PMID- 21615829 TI - In vitro production of porcine embryos: current status, future perspectives and alternative applications. AB - The pig is considered to be a suitable source of cells and organs for xenotransplants, as well as a transgenic animal to produce specific proteins, given the biological similarities it shares with human beings. However, the in vitro embryo production system in pigs is inefficient compared with those in other mammals, such as cattle or mice. Although numerous modifications have been applied to improve the efficiency of in vitro embryo production systems in pigs, not much progress has been made to overcome the problem of polyspermy, and low developmental ability due to insufficient cytoplasmic abilities of in vitro matured oocytes and improper culture conditions for the in vitro produced embryos. Recent achievements, such as the establishment of chemically defined medium and utilization of 'zona hardening' technique, have gained some success. However, further research for the reduction of polyspermy and detrimental effects of the culture systems in pigs is still needed. PMID- 21615830 TI - Selecting the desirable method for predicting 305-day lactation yields in Mongolia. AB - In Mongolia, milk production must be improved both quantitatively and qualitatively, and we must find the most desirable method for predicting 305-day milk yields for cows in Mongolia. Therefore, the predicted 305-day milk yield from the test interval method (TIM), multiple-trait prediction (MTP) and the random regression model (RRM) were compared. The daily milk production records during 305 days for cows calving from 1986 to 2007 from the National Livestock Breeding Center in Japan, and the test day records every month for cows calving from 1985 to 2005 from the Hokkaido Dairy Milk Recording and Testing Association were used. Wilmink's model for the average of lactation curve in MTP was adopted. A cubic Legendre polynomials and the parameters of Wilmink's function were used for RRM. The results suggested that RRM was the most desirable method for predicting 305-day milk yields in Mongolia because estimates by RRM were the most accurate when using over four records. Further analyses will be required to examine the properties of RRM when predicting 305-day milk yields using test day records in Mongolia. PMID- 21615831 TI - Homozygosity of single nucleotide polymorphisms in the 3' region of the canine estrogen receptor 1 gene is greater in Toy Poodles than in Miniature Dachshunds and Chihuahuas. AB - Differences in the distribution of single nucleotide polymorphisms (SNPs) and haplotypes in the estrogen receptor alpha gene (ESR1) were examined in Miniature Dachshunds (n = 48), Chihuahuas (n = 20) and Toy Poodles (n = 18). Five DNA fragments located in the 40-kb region at the 3' end of ESR1 were amplified by polymerase chain reaction and were directly sequenced. We compared allele, genotype and estimated haplotype frequencies at each SNP in the 3' end of ESR1 for these three breeds of small dog. The frequency of the major allele and the genotype frequency of the major allele homozygotes, were significantly higher in Toy Poodles for five SNPs (SNP #5, #14-17) than in Miniature Dachshunds, and significantly higher in Toy Poodles than Chihuahuas for three SNPs (SNP #15-17). A common haplotype block was identified in an approximately 20-kb region encompassing four SNPs (SNPs # 14-17). The frequencies of the most abundant estimated haplotype (GTTG) and GTTG homozygotes were significantly higher in Toy Poodles than in the other two breeds. These results imply that homozygosity for the allele, genotype and haplotype distribution within the block at the 3' end of ESR1 is greater in Toy Poodles than in Miniature Dachshunds and Chihuahuas. PMID- 21615832 TI - Genetic association between GHSR1a 5'UTR-microsatellite and nt-7(C>A) loci and growth and carcass traits in Japanese Black cattle. AB - We carried out a genetic association study between five nucleotide polymorphisms (5'UTR microsatellite ((TG)(n)), nt-7(C>A), L24V, DelR242 and Intron 1 microsatellite) of the GHSR1a gene and growth and carcass traits in 1285 steers sired by 117 Japanese Black bulls in a progeny testing program. We report herein, a significant association between the 5'UTR microsatellite and nt-7(C>A) loci and growth and carcass traits. We also propose a translational hypothesis that the association is due to differences in the secondary structure of GHSR1b mRNA (the non-spliced type with the 5'UTR microsatellite) among the GHSR1a gene haplotypes. Furthermore, we predicted the potential increase in profitability due to increased carcass weight in cow-calf fattening enterprises through planned matings based on DNA testing of the 5'UTR microsatellite. Statistical analysis revealed that the 5'UTR microsatellite locus had a significant additive effect on carcass weight (CW) and average daily gain (ADG), but not on beef marbling score (BMS). One of the four major microsatellite alleles (19-TG allele) with an allele frequency of 0.145, had a significantly (P < 0.0007) desirable effect on CW and ADG. We concluded that the 19-TG allele could potentially be economically useful nucleotide markers for growth and carcass traits in Japanese Black cattle. PMID- 21615833 TI - Effect of DNA polymorphisms related to fatty acid composition in adipose tissue of Holstein cattle. AB - Fatty acid composition of adipose tissue has been recognized as an important carcass trait because of its relationship with eating quality such as favorable beef flavor and tenderness. Therefore, we investigated the effects of genetic polymorphisms of liver X receptor, alpha (LXR), stearoyl-CoA desaturase (SCD), Fatty acid synthase (FASN), and Fatty acid binding protein 4 (FABP4) on fatty acid composition in intramuscular fat tissue of Holstein steers. The major allele frequencies were 0.705 in SCD, 0.518 in FABP4, 0.888 in FASN, and 0.984 in LXR. Genotyping of SCD showed significant effect on C14:0, C14:1, C18:0 and saturated fatty acid (P < 0.05). In addition, the result suggested that SCD genotype possibly had effect on composition of C18:1 and monounsaturated fatty acid. Genotype of FABP4 had significant effect on composition of C16:0. Effect of LXR genotypes could not be analyze because of extremely biased genotype frequencies. Our results suggest that genotypes of SCD and FABP4 may in part affect meat quality in Holstein. PMID- 21615834 TI - The addition of calcium ion chelator, EGTA to thawing solution improves fertilizing ability in frozen-thawed boar sperm. AB - In our previous study, seminal plasma effectively suppressed the induction of sperm to capacitation-like status and acrosome loss during the thawing process. However, because boar seminal plasma is contaminated with various kinds of bacteria and/or viruses, it is necessary to develop a thawing solution without animal-derived materials. In this study, we focused on the increase of intracellular Ca(2+) ([Ca(2+)](i)) in sperm after thawing and the negative effects of sperm qualities. After thawing, the fluorescent intensity of [Ca(2+)](i) indicator, Fluo-3/AM, and the level of phosphorylated tyrosine residue of protein were increased in the sperm. Next, we investigated whether the addition of Ca(2+) chelators (ethylenediaminetetraacetic acid (EDTA) and ethyleneglycoltetraacetic acid (EGTA)) improved post-thawed sperm motility. When the frozen-thawed sperm were treated with 6 mmol/L EDTA + 6 mmol/L EGTA, sperm motility was significantly increased as compared with control (6 mmol/L EDTA alone) at all incubation periods (P < 0.05). The combinational treatment significantly suppressed the elevation of [Ca(2+)](i) and the tyrosine phosphorylation, which improved the acrosomal status and fertilizing ability in vitro. Furthermore, when the thawing method was applied for artificial insemination, the fertilization rate was significantly higher than control (P < 0.05, 33% vs. 82%). Thus, we conclude that the addition of EDTA + EGTA to thawing solution is a beneficial tool for artificial insemination using frozen-thawed boar sperm. PMID- 21615835 TI - Mouse embryo co-culture with autologous cumulus cells and fetal development post embryo transfer. AB - This study was conducted to examine the potential for implantation and sustainable fetal development of mouse embryos cultured from the pronuclear to blastocyst stage. Pronuclear embryos from ICR mice (Harlan Sprague-Dawley) were cultured in Sydney IVF sequential media (Cook) to the blastocyst stage in medium only or co-cultured with autologous cumulus cells. We also experimented with co culture in 100 uL drops. Drop co-culture produced blastocyst formation rates with a mean of 47.0%, which was significantly higher (P < 0.05) compared to embryos cultured in identical culture conditions except without cumulus cells at 27.3%. Blastocysts obtained in vitro in Cook medium only and co-cultured in Cook medium with cumulus cells were transferred to pseudopregnant females of ICR strain. The day of blastocyst transfer into surrogate females was designated as post-transfer of blastocyst day 1 (PT 1). The implantation and fetal development was compared to embryo transfer of in vivo derived blastocysts, which served as controls. There were no statistical differences for implantation and fetal development rates for blastocysts cultured in vitro in either Cook medium only or co-culture in Cook medium with cumulus cells compared to in vivo-derived blastocysts. The advantage of the co-culture system is in generating more blastocysts available for transfer. PMID- 21615836 TI - Detection of the EGFP sequence in breast muscle of 3-year-old chicken after transfection using sonoporation. AB - In our continuing effort to generate transgenic chickens, sonoporation was chosen to insert an exogenous gene into the chicken genome. An EGFP expression vector (pCAG-EGFPac) and microbubbles were injected into the central disc of stage-X blastoderm or the germinal crescent of stage-4 embryos, followed by ultrasonic vibration. Nineteen chicks out of 108 treated embryos hatched, six females and six males out of these 19 chicks grew to sexual maturity and two females and three males lived for 3 years. Genomic DNA from 17 out of 35 gonads from embryos and chicks that died before sexual maturity was EGFP-positive by PCR. No EGFP sequence was detected in the genomic DNA of 322 embryos from six sexually mature females and the semen from four sexually mature males by PCR. When genomic DNA was obtained from various tissues of five 3-year-old chickens, the EGFP sequence was amplified from the genomic DNA of the breast muscle of a female (No. 85). The above sequence was subjected to DNA sequencing and verified to be the EGFP sequence. These results showed that sonoporation is an effective tool for the transduction of exogenous genes into chicken embryos for the generation of transgenic chickens. PMID- 21615837 TI - Fatty acid composition of ruminal bacteria and protozoa, and effect of defaunation on fatty acid profile in the rumen with special reference to conjugated linoleic acid in cattle. AB - Objectives of this study were to compare fatty acid (FA) composition of ruminal bacterial (B) and protozoal (P) cells, and to investigate effect of protozoa on FA profile in the rumen of cattle. Three cows were used to prepare ruminal B and P cells. Four faunated and three defaunated cattle (half-siblings) were used to study effect of protozoa on ruminal FA profile. Proportions of C16:0 and C18:0 in total fatty acids in B cells were 20.7% and 37.4%, whereas those in P cells were 33.4% and 11.6%, respectively. Proportions of trans-vaccenic acid (VA) and cis-9, trans-11 conjugated linoleic acid (CLA) in B cells were 3.9% and 1.0%, and those in P cells were 5.5% and 1.6%, respectively, being higher in P cells. Proportions of C18:1, C18:2 and C18:3 in P cells were two to three times higher than in B cells. Proportions of unsaturated fatty acids, VA and CLA in B cells of faunated cattle were higher than those of defaunated. VA and CLA in the ruminal fluid of faunated were also 1.6 to 2.5 times higher than those of defaunated. This tendency was similar for cell-free fraction of ruminal fluid. These results indicate that protozoa contribute greatly in VA and CLA production in the rumen. PMID- 21615838 TI - Rumen microbial response in production of CLA and methane to safflower oil in association with fish oil or/and fumarate. AB - Supplementation effect of fish oil and/or fumarate on production of conjugated linoleic acid (CLA) and methane by rumen microbes was examined when incubated with safflower oil. One hundred and twenty milligrams of safflower oil (SO), safflower oil with 24 mg fish oil (SOFO), safflower oil with 24 mmol/L fumarate (SOFA), or safflower oil with 24 mg fish oil and 24 mmol/L fumarate (SOFOFA) were added to the 90 mL culture solution. The culture solution was also made without any supplements (control). The SOFA and SOFOFA increased pH and propionate (C3) compared to other treatments from 3 h incubation time. An accumulated amount of total methane (CH(4) ) for 12 h incubation was decreased by all the supplements compared to control. The concentrations of c9,t11CLA for all the incubation times were increased in the treatments of SOFO, SOFA and SOFOFA compared to SO. The highest concentration of c9,t11CLA was observed from SOFOFA among all the treatments at all incubation times. Overall data indicate that supplementation of combined fumarate and/or fish oil when incubated with safflower oil could depress CH(4) generation and increase production of C(3) and CLA under the condition of current in vitro study. PMID- 21615840 TI - Relationships between urine pH and electrolyte status in cows fed forages. AB - Data of 20 balance measurements from Holstein dairy cows and urine samples from 24 Japanese Black beef cows were collected to evaluate the relationships between urine pH and electrolyte status in cows fed forages. The ratio of forages in the diet was 70-100% in dairy cows and beef cows were fed Italian ryegrass silage and wheat bran. Mean urine pH in dairy cows was 8.10, ranging from 7.27 to 8.71, and that in beef cows was 7.73, ranging from 7.42 to 8.12. There were positive correlations between urine pH and urinary K contents (P = 0.0012) or K intake (P = 0.019) in dairy cows, although plasma Na, Cl and K had no effect on urine pH. There was a weak negative correlation (P = 0.039) between urine pH and urinary Na content in dairy cows. However, there were no significant correlations between urine pH and urinary Na, Cl and K contents in beef cows. These results indicate that the concentrated urinary K due to the increased K intake may directly enhance urine pH in dairy cows fed mainly forages. PMID- 21615839 TI - Effect of ensiled hop (Humulus lupulus L.) residues on plasma acetate turnover rate in sheep. AB - An isotope dilution method using [1-(13)C]sodium acetate was applied to determine the effect of feeding ensiled hop (Humulus lupulus L.) residues on plasma acetate turnover rate in six adult crossbred sheep. The sheep were fed 63 g/kg body weight (BW)(0.75)/day of either mixed hay of orchardgrass (Dactylis glomerata L.) and reed canarygrass (Phalaris arundinacea L.) and round bale silage at 3:1 ratio (Hay-diet), or another where round bale silage was replaced by ensiled hop residues (Hop-diet) with a crossover design each of a 3-week period. The isotope dilution method was performed on day 21 of each dietary treatment. Dry matter digestibility was similar between diets, and nitrogen (N) digestibility was lower (P = 0.001) for Hop-diet than Hay-diet. However, N retention did not differ between diets. Plasma acetate concentration was lower (P = 0.04) for Hop-diet than Hay-diet, and the turnover rate of plasma acetate did not differ between diets. Plasma concentration of lactate and non-esterified fatty acids were similar between diets. Hop-diet was found almost comparable to Hay-diet on plasma acetate turnover rate in the present experimental conditions. Therefore, it could be concluded that hop residues partially could be used as an alternative to traditionally used round bale silage for rearing sheep. PMID- 21615841 TI - Effects of corn dried distiller's grains with solubles and enzyme premix supplements on growth performance, carcass characteristics and meat quality parameters in finishing pigs. AB - The objective of the present study was to investigate the effects of corn dried distiller's grains with solubles (DDGS) and enzyme premix (mannanase + phytase) supplementation on the growth performance, carcass and meat quality parameters in finishing pigs. Sixty hybrid pigs (L * LW * D) with initial weight of 63.92 +/- 1.50 kg were used in a 3 * 2 factorial design with main effects of DDGS levels (0, 10 and 20%) and enzyme premix levels (0% vs. 0.14%). Average daily gain (ADG, P < 0.01) and average daily feed intake (ADFI, P < 0.05) were decreased due to an increased level of DDGS additive while the feed conversion ratio was improved (P < 0.05) by adding enzyme premix. The diet cost/gain (won/kg) was saved (P < 0.01) due to an increased level of DDGS additive. There were no significant differences in carcass characteristics and meat quality parameters of Longissimus dorsi muscle by DDGS level and enzyme premix. Palmitoleic acid, oleic acid and monounsaturated fatty acid (MUFA) decreased (P < 0.05) according to DDGS level. The results indicate that DDGS may be used in feeds for finishing pig as a replacement of corn and soybean meal without affecting their carcass characteristics and meat quality. PMID- 21615842 TI - Effect of dietary inclusion of sorghum milling waste on growth response, nutrient utilisation, gut characteristics and cecal microflora of weaner rabbits. AB - Growth response, nutrient digestibility and cecal microflora of 80 male, mixed breed weaner rabbits fed with varying dietary inclusions of sorghum milling waste (SMW) was investigated. Four experimental diets were formulated such that SMW was included at 0 (control), 100, 200 and 300 g/kg, respectively. Each dietary treatment was performed on 20 rabbits. Feed intake increased (P < 0.05) while final live weight and feed conversion ratio of rabbits decreased (P < 0.05) following increased dietary inclusion of SMW. Rabbits fed with 100 and 200 g/kg SMW had similar feed conversion ratios, weight gain, crude fiber, dry matter and crude protein digestibility values. Rabbits fed with 300 g/kg SMW recorded the lowest (P < 0.05) hot carcass weight, dressing percentage and rack weight. Similar dressing percentage and rack weight were recorded for rabbits fed with control diet, 100 and 200 g/kg SMW. The weight of cecal content increased (P < 0.05) with increased dietary inclusion levels of SMW. Rabbits fed with 300 g/kg SMW recorded the lowest (P < 0.05) coliform and lactobaccillus counts. Dietary inclusion of up to 200 g/kg SMW supported improved growth response and carcass yield without imposing any detrimental effect on cecal microflora. PMID- 21615843 TI - Effect of calcitonin on adrenocorticotropic hormone secretion stimulated by corticotropin-releasing hormone in the hen anterior pituitary. AB - The presence of a receptor for calcitonin (CT) and the effect of chicken CT (cCT) on adrenocorticotropic hormone (ACTH) secretion stimulated by rat/human corticotropin-releasing hormone (rhCRH) in the hen anterior pituitary were studied. The specific [(125)I]cCT binding component was present in the plasma membrane of hen anterior pituitary and this binding component had properties of a receptor which has binding specificity to cCT, reversibility, saturable binding, high affinity and limited capacity. When anterior pituitary cells were incubated in vitro, cCT increased the maximal secretion of chicken ACTH stimulated by rhCRH. These results suggest that CT may act directly on the anterior pituitary via its receptor binding and enhances the ACTH secretion by CRH. PMID- 21615844 TI - Influence of shearing on oxidative stress and some physiological parameters in ewes. AB - The aim of the present study was to evaluate the effect of shearing on physiological and oxidative stress parameters in ewes. Twenty Comisana ewes were used and divided into two groups. Ten ewes were left unshorn as a control group (Group A) and 10 ewes were shorn (Group B). All measurements were taken before and after shearing, and repeated 8 h after shearing and 1, 2, 3, 4, 5, 10 and 15 days after shearing. Reactive oxygen species (dROMs), antioxidant barrier (oxy adsorbent), thiol antioxidant barrier (SHp) and packed cell volume (PCV) were assessed in blood samples collected by means of jugular venipuncture. Rectal temperature (RT), respiratory rate (RR) and heart rate (HR) were also measured. Two-way repeated measures analysis of variance (ANOVA), followed by Bonferroni's test, was used for the assessment of significant effects due to shearing and time. The statistical analysis showed significant increases (P < 0.01) of dROMs, oxy-adsorbent, SHp, and a significant decrease (P < 0.01) of RT and RR associated with time and shearing. Our results indicate that shearing causes a change in the ewe's homeostatic balance that leads to oxidative stress. PMID- 21615845 TI - Economic effects of using sexed semen in Japanese dairy herds. AB - Dairy farm agricultural income should increase when sexed semen is used instead of unsexed semen. This study simulated three scenarios: using unsexed semen (N), using X sperm of Holstein and unsexed semen of Japanese Black (H), and using X sperm of Holstein and Y sperm of Japanese Black (HJ). Two management scenarios were considered: conventional management (CONV); and the use of Japanese Black semen with surplus cows to produce F(1) s where sufficient replacement female Holstein calves were obtained for maintenance (MAINT). In CONV, the agricultural incomes of H and HJ were 610,000-2,400,000 yen higher than that of N. The agricultural income increased when the difference in the prices between Holstein males and average F(1) male/female calves was 34,000-50,000 yen, that between Holstein male and female calves was 80,000-110,000 yen, and that between F(1) male and female calves was 50,000-90,000 yen. Therefore, the agricultural income can be increased by using sexed semen to select calves of a more valued breed and sex. PMID- 21615846 TI - Lessons from a transplant patient with diarrhea, cryptosporidial infection, and possible mycophenolate mofetil-associated colitis. AB - Diarrhea in a transplant recipient may be caused by infection, metabolic problems, or adverse drug effects. The immunosuppressive drug most frequently associated with diarrhea in transplant recipients is mycophenolate mofetil (MMF). We present the case of a patient with 2 potential explanations for diarrhea lasting several weeks, which occurred years after liver transplantation. Whereas stool samples were positive for cryptosporidia, the histopathological findings were compatible with MMF colitis. However, diarrhea resolved after treatment of cryptosporidial infection, despite continued MMF medication. This case shows that histopathological findings of MMF colitis may be misleading and do not prove that diarrhea is drug induced. PMID- 21615847 TI - Persistence of influenza vaccine-induced antibodies in lung transplant patients between seasons. AB - BACKGROUND: Immunization policy-making bodies advised against immunizing too early before the influenza season because vaccine-specific antibody may wane before the end of the influenza season. Lung transplant patients are included in the group of high-risk patients for whom this recommendation had been made. We hypothesized that immunosuppressed lung transplant patients would maintain protective concentrations of influenza antigen-specific antibodies between seasons. METHODS: As part of a planned 5-year study of influenza vaccine responses in lung transplant patients, we measured influenza antibody concentrations by hemagglutination inhibition assay before influenza immunization annually. The fraction of lung transplant patients who maintained seroprotective levels (>= 40 hemagglutination units) approximately 11 months from last season immunization was calculated. Antibody concentrations and response rates in lung transplant patients were compared with healthy individuals and those waiting for lung transplantation. RESULTS: The majority of lung transplant patients maintained seroprotective influenza antigen-specific antibody concentrations for approximately 11 months after immunization. Seroprotection rates varied greatly with influenza antigens (healthy 68-100%, pretransplant 44-100%, transplant 64 100%), and were similar when groups were compared. More than 70% of lung transplant patients maintained seroprotective antibody concentrations to 10 of 11 vaccine antigens. CONCLUSION: Seroprotective influenza antibody concentrations are maintained at very high rates among immunosuppressed lung transplant patients and depend more on the vaccine virus than the immunostatus of the vaccine recipient. Early seasonal influenza immunization of lung transplant patients is appropriate. PMID- 21615848 TI - Onset and complications of varicella zoster reactivation in the autologous hematopoietic cell transplant population. AB - BACKGROUND: Varicella zoster virus (VZV) infections are a common complication in patients receiving autologous or allogeneic hematopoietic cell transplant (HCT). Recent guideline revisions suggest extending VZV prophylaxis to 1 year after autologous HCT. We retrospectively evaluated reactivation at our center, before implementation of extended acyclovir prophylaxis, to determine onset and outcome in the autologous HCT population. METHODS: Inclusion criteria consisted of adult patients who received an autologous HCT with documentation for at least 1 year post transplant. Those excluded from review were patients who received acyclovir prophylaxis for >30 days post transplant or subsequently received an allogeneic transplant within 1 year. For patients in whom reactivation occurred, the severity of infection, the timing of onset, treatment of the reactivation, and any complications were recorded. RESULTS: In the final analysis, 56 patients were assessed. Reactivation of zoster occurred in 16% of recipients with a median onset of 4.5 months post transplant. Complications that were observed include postherpetic neuralgia, severe pain, scarring, and motor weakness. Two patients required hospitalization for treatment, with 1 patient requiring 6 months of rehabilitation for motor weakness following the infection. CONCLUSIONS: Our study revealed a 16% incidence of VZV reactivation in our autologous HCT population. The onset of these occurrences ranged from 2 to 10 months post transplant, with significant VZV-associated complications. We consider VZV reactivation a serious concern in the autologous transplant setting, requiring extended prophylaxis. PMID- 21615849 TI - Dynamic prediction of metastases after radical prostatectomy for prostate cancer. AB - What's known on the subject? and What does the study add? One of two problems plagues virtually are existing post-prostatectomy prediction tools: either (1) they predict PSA recurrences (which are of unclear importance) or (2) the predictions they make are anchored at the date of surgery and are not updated based on how patients evolve over the postoperative years. Our prediction tool is a significant improvement over existing prediction tools in that it predicts the development of metastases which is a very important clinical endpoint that indicates incurable prostate cancer. Additionally, our tool allows for updated predictions at any point following radical prostatectomy by considering commonly available postoperative information (postoperative serum PSA and use of adjuvant therapies) to modify its risk predictions. The net result is a dynamic tool that renders clinically relevant predictions that change as the patient's clinical status changes throughout the postoperative course. OBJECTIVE: * To develop a dynamic algorithm that predicts the risk of metastases from any time point after radical prostatectomy (RP). PATIENTS AND METHODS: * The study cohort consisted of 5741 RP patients who were treated from 1990-99. * Patients were grouped into one of four clinical states at follow-up: State(1) , prostate-specific antigen (PSA) undetectable; State(2), PSA 0.15-0.39 ng/mL; State(3), PSA >=0.4 ng/mL; and State(4), previous androgen deprivation or radiation therapy. * Follow-up epochs (alive and at risk of systemic progression) at 0, 2, 4 and 6 years post-RP, cumulative incidence curves and multistate Cox models were used to assess the risk of metastases over the ensuing 5-year interval. * Gleason score, seminal vesicle and surgical margin involvement, and PSA variables were evaluated as predictors. RESULTS: * Median follow-up was 11.7 years, with 4411, 4256 and 3983 patients followed with PSA at 2, 4 and 6 years, respectively. * In total, 287 metastatic events occurred and the 5-year risk of metastasis was 0.4%, 2.1%, 8.7% and 12.6% for men in States 1, 2, 3 and 4, respectively. * Independent predictors of metastasis by group included seminal vesicle involvement (all groups), Gleason score (groups 1, 3 and 4), current PSA (groups 3 and 4) and maximum past PSA (group 4). CONCLUSIONS: * We present a web-based prognostic tool for patients undergoing RP that is valid at many time points after surgery. * Our tool predicts the development of metastases. PMID- 21615850 TI - Clinically unconfirmed positive urinary cytology: diagnostic implications and oncological outcomes. AB - OBJECTIVE: *To evaluate the natural history of patients presenting with positive urinary cytology in the absence of clinically identifiable disease. PATIENTS AND METHODS: *A surgical database was queried to identify 48 patients with positive urinary cytology of undetermined source. *All patients underwent a thorough urological evaluation consisting of random bladder biopsies, lateral montanal prostate biopsies, bilateral retrograde pyelography and selective urine cytology sampling from each ureter. *Time from presentation to definitive diagnosis was estimated using the Kaplan-Meier method. RESULTS: *In total, 27 patients (56%) had a history of bladder cancer and 21 (44%) were evaluated for irritative voiding symptoms or haematuria. *At a median follow-up of 31 months, the source of the positive cytology was identified in 32 patients (67%): in 29 (61%) as bladder cancer, in two patients (4%) as upper tract tumour and in one patient (2%) as transitional cell carcinoma of the prostate; 29/32 (91%) had high-grade disease and the predominant clinical stage (53%) was carcinoma in situ. *The median (interquartile range) interval from presentation to diagnosis was 19 (8 22) months, and was significantly shorter in naive patients than in those with a previous history of bladder cancer (13 vs 27 months, P < 0.05). *Disease-free survival with an intact bladder at 4 years was 74% (95% CI, 63-85). CONCLUSIONS: *Unconfirmed positive urine cytology often predates the development of high-grade urothelial carcinoma. *The bladder is most commonly involved. *The time to diagnosis is generally protracted, although the long0term outcome appears to be favourable. PMID- 21615851 TI - Impact of urethral ultrasonography on decision-making in anterior urethroplasty. AB - OBJECTIVE: To assess the impact of urethral ultrasonography on decision-making in complex anterior urethral reconstructions. PATIENTS AND METHODS: This was an institutional review board approved retrospective review of 500 patients receiving anterior urethroplasty from 1985 to 2007. In all, 232 patients met the inclusion criteria of documented preoperative clinical assessment, retrograde urethrogram (RUG), urethral ultrasonogram, and measured operative stricture length and dimensions. At the time of surgery, intraoperative urethral ultrasonography was performed. A preoperative planned reconstructive approach was compared with the reconstructive procedure performed to determine how often urethral ultrasonography impacted operative decision-making. RESULTS: Of the 232 patients in our review, 115 (50%), 88 (38%) and 16 (7%) received respectively anastomotic, buccal mucosa graft and fasciocutaneous flap urethroplasties. Intraoperative ultrasonography changed surgical approach in 44 patients (19%), and was integral in deciding between two equally possible approaches in an additional 61 patients (26%). In patients whose approach was changed from anastomotic urethroplasty to an onlay urethroplasty, mean RUG length was 2.0 cm while mean ultrasound length was significantly longer at 3.4 cm (P= 0.02). When the surgical approach was changed from an onlay urethroplasty to an anastomotic urethroplasty, mean RUG length was 2.7 cm compared with a significantly shorter intraoperative ultrasonography length of 1.9 cm (P < 0.005). CONCLUSIONS: Ultrasonography of the anterior urethra directly influenced our reconstructive operative approach in 45% of patients. It is a simple, reliable procedure that adds valuable clinical insight and objective radiographic data to help select the optimal anterior urethral reconstructive approach. PMID- 21615852 TI - Distal urethroplasty for isolated fossa navicularis and meatal strictures. AB - OBJECTIVE: * Urethral strictures located in the fossa navicularis are common and are often managed with meatotomy or meatoplasty. * Few data have described the outcomes for men after urethroplasty or patient satisfaction following these procedures. METHODS: * In all, 93 men at two different institutions underwent surgical repair of distal urethral stricture disease using meatotomy (73) or meatoplasty (20), with 13/20 (65%) of the latter group undergoing substitution urethroplasty. * In patients with lichen sclerosus (LS), all involved tissue was excised prior to reconstruction. * In a subset of men undergoing meatotomy, patient satisfaction was evaluated by questionnaire. RESULTS: * Average clinical follow-up for men undergoing distal urethroplasty was 61 months. * Successful reconstruction requiring no further intervention occurred in 84% of men overall. Subgroup analysis revealed success in 87% of men with meatotomy, 75% with meatoplasty and 66% with substitution urethroplasty. * Men with LS had a significantly greater rate of stricture recurrence (20.5% vs 7.5%, P= 0.04). * Of the subset of men who completed a patient-based questionnaire 84% reported they were either satisfied or very satisfied with the results of their meatotomy. CONCLUSIONS: * We report the success of distal urethral stricture management. * Meatal strictures may be approached successfully in a stepwise manner progressing from meatotomy to meatoplasty for longer recurrent strictures, with a high overall success rate for meatotomy. * Although substitution grafts may be useful for men with longer distal strictures and those with LS, the risk of recurrence was significantly higher in this cohort. PMID- 21615853 TI - Trends in adverse events of benign prostatic hyperplasia (BPH) in the USA, 1998 to 2008. AB - OBJECTIVE: To determine if the adverse events (AEs) of benign prostatic hyperplasia (BPH) have declined in tandem with increased use of oral therapy. MATERIALS AND METHODS: We used the Nationwide Inpatient Sample, a 20% sample of USA community hospitals, weighted to estimate national numbers to characterize the prevalence of AEs of BPH from 1998 to 2008. We calculated the age-adjusted prevalence of BPH and associated conditions and analyzed prevalence trends with regression modelling. RESULTS: Of 134 million estimated eligible discharges during the study period, 7,464,730 (5.6%) had either a primary or secondary diagnosis of BPH. The age-adjusted prevalence of BPH among all hospitalizations, irrespective of primary diagnosis, increased from 4.3% to 8% (P < 0.001) during the study period. The age-adjusted prevalence of BPH as a primary diagnosis decreased from 0.88% to 0.48% (P < 0.001). Discharges for BPH surgery decreased 51% (odds ratio [OR] 0.49, 95% confidence interval [CI] 0.45-0.54, P-trend <0.001) over time. Discharges for primary BPH with acute renal failure increased >400% (OR 4.28, 95% CI 3.22-5.71, P-trend <0.001). There were no significant changes in discharges for primary BPH with urinary retention (P-trend = 0.636), bladder stones (P-trend = 0.117), or urinary infection (P-trend = 0.101) over time. CONCLUSIONS: Increased hospitalizations for BPH with acute renal failure and stable hospitalizations for other AEs of BPH indicate that severe AEs of BPH persist despite widespread use of oral therapies in the USA. Further studies are needed to explain these trends. PMID- 21615854 TI - Impact of docetaxel-based chemotherapy on quality of life of patients with castration-resistant prostate cancer: results from a prospective phase II randomized trial. AB - What's known on the subject? and What does the study add? Data on quality of life during docetaxel treatment in castration resistant prostate cancer was mainly provided by SWOG and TAX327 trials. In the TAX327 trial biochemical response and pain predicted survival, whereas quality of life outcomes did not. In the present study, there were no statistically significant changes in the quality of life scales during treatment except in the case of patients receiving docetaxel and estramustine, who experienced a significant decrease in pain. Our data seem to suggest that patients with a better baseline quality of life (and consequently with fewer symptoms) are more likely to achieve a biochemical response. OBJECTIVES: * To assess quality of life (QoL) outcomes and pain changes in patients affected by castration-resistant prostate cancer enrolled in a phase II randomized trial of 3-week docetaxel (DOC)-based chemotherapy. * To provide further data to clarify the conflicting published data concerning the impact of DOC on the patients' QoL. PATIENTS AND METHODS: * QoL outcomes were assessed using the European Organisation for the Research and Treatment of Cancer QLQ-C30 questionnaire. * Pain changes were evaluated by means of the Brief Pain Inventory at baseline and after every two DOC courses. * The patients completing at least two questionnaires (at baseline and before the third course) were considered evaluable. RESULTS: * In all, 59 patients were evaluable. * Asymptomatic patients and responders had a better baseline QoL than symptomatic patients and non responders. * There were no statistically significant changes in the QLQ-C30 scales during treatment except in the case of patients receiving DOC and estramustine, who experienced a significant decrease in pain. * There was a progressive improvement in the mean intensity and interference scores of the Brief Pain Inventory. CONCLUSIONS: * Our data confirm that QoL is generally maintained during chemotherapy. * There is a substantial reduction in pain. * Our results also suggest that baseline QoL may predict treatment response. PMID- 21615855 TI - Early ultrasound-guided neurolysis for pain management in gastrointestinal and pelvic malignancies: an observational study in a tertiary care center of urban India. AB - Patients with advanced gastrointestinal and pelvic malignancies commonly present with pain of varying severity. In a majority of these patients, pain can be effectively managed using an integrated systemic pharmacological approach with oral morphine being the cornerstone of treatment. However, with escalating doses, intolerable side effects of oral morphine may lead to patient dissatisfaction. When oral pharmacotherapy fails to adequately address the issue of pain or leads to insufferable side effects, neurolytic blocks of the sympathetic axis are usually used for pain alleviation. As these blocks may reduce oral analgesic requirement, a reevaluation of their timing is merited. This article presents our hospital-based in-patient palliative care unit experience with early ultrasonography-guided neurolysis of celiac plexus, superior hypogastric plexus and ganglion impar. Of the 44 patients we studied, 20 underwent celiac plexus neurolysis, 18 superior hypogastric plexus neurolysis, and 6 ganglion impar neurolysis. Their pain was being managed with oral morphine before neurolysis, but only 11.4% patients required oral morphine for satisfactory pain control, 2 months after neurolysis. The mean Visual Analog Scale score before block placement was 5.64 +/- 0.69 and fell to 2.25 +/- 1.33 at 2 months post neurolysis (P < 0.001). We suggest that bedside ultrasonography-guided sympathetic axis neurolysis may be employed early in patients with incurable abdominal or pelvic cancer. Its use as a first-line intervention for achieving pain control with minimal complications warrants further consideration and investigation. PMID- 21615856 TI - Epidural steroid and clonidine for chronic intractable post-thoracotomy pain: a pilot study. AB - BACKGROUND: Chronic post-thoracotomy pain is relatively common after major thoracic surgery. The primary results of a pilot study using thoracic epidural steroid and clonidine injection to treat chronic intractable post-thoracotomy pain are presented. METHODS: Twenty-one patients with intractable post thoracotomy pain participated in the study. Thirteen patients received thoracic epidural injection of a mixture of 150 MUg clonidine and 80 mg of methylprednisolone acetate diluted in 8 mL 0.5% lidocaine. Eight patients continued with comprehensive medical management and served as a control group. A visual analog scale (VAS) for pain was recorded before treatment, 30 minutes after the epidural injection and before discharge, at 3 weeks and 6 months. Pain, sleep disturbances, appetite changes and daily activity, as well as the incidence of complications were recorded. The need for opioid rescue medications was recorded. RESULTS: Twelve of 13 patients in the injection group reported improvement (> 50% reduction of pain) at 3 weeks and 6 months following the injection. Allodynia improved in all injection group patients compared to four of eight in the control group. Sleep disturbance, appetite changes and daily activity were improved in the injection group. The number of patients requiring opioid rescue medications was reduced from 61.5% to 15.3% during the 6-month duration of study. Injection caused transient hypotension in 46.2% of patients. Mild sedation was noted in 30.7% of patients receiving injection; 15.3% of the patients had localized back pain at the site of injection. DISCUSSION: Our preliminary data suggest possible efficacy of thoracic epidural steroid and clonidine mixture in the treatment of chronic post-thoracotomy pain. No serious adverse effects were noted in this pilot study. PMID- 21615858 TI - The story of the Texas Pain Society: formation and function of a regional pain society. AB - The idea of forming a Texas Pain Society came to the Founders in 1987 due to disparity and deficiencies in the practice of pain management in the United States and, in particular, the State of Texas. The Founders considered very carefully the implication of forming such a society. They diligently mapped out the mission and goals of the Texas Pain Society in those early formative years. This report is the history of Texas Pain Society as the activities unfolded from 1989 to 2011. The reader may question why there is a need to tell such a story. We believe strongly that, with disparities of standards of practice in pain medicine and poor recognition of advances in pain management, this scenario is quite common in many states and countries. The practitioners of pain management in these regions certainly must have considered getting together and forming a consensus on the standards of practice in their communities. This historical report of the Texas Pain Society provides the relevant information necessary and the efforts to be made for a society's mission to achieve its goals and have an ongoing impact in its own region. We hope that we have shed some light on a process for the formation of a regional pain society such as ours. PMID- 21615857 TI - Health care costs in patients with fibromyalgia on pregabalin vs. duloxetine. AB - BACKGROUND: The economic burden associated with fibromyalgia in the U.S. is substantial. The objective of this study was to compare changes in health care costs in fibromyalgia patients initiated on pregabalin and duloxetine in real world settings. METHODS: Patients (>= 18 years old) initiating pregabalin or duloxetine between June 1, 2007 and December 31, 2008 were identified using a U.S. managed care database. Patients were selected if they had >= 2 medical claims for fibromyalgia (ICD-9-CM, 729.1) at least 90 days apart or >= 1 claim for fibromyalgia followed within 30 days by a pharmacy claim for pregabalin. The date of the first pregabalin or duloxetine prescription was defined as the index date, and continuous enrollment for 6-month pre- and postindex periods was required. RESULTS: A total of 1,616 pregabalin and 207 duloxetine patients were identified. Treatment differences between pregabalin and duloxetine in the pre /postindex change in mean [SD] all-cause total health care costs ($1,307 [16,747] vs. -$158 [17,337]; P = 0.24) or fibromyalgia-related total health care costs ($584 [3,834] vs. $759 [2,133]; P = 0.32) were not significant. Multivariate analysis using difference-in-differences models showed no significant difference in all-cause costs (mean cost ratio = 1.05, 95% CI: 0.84 to 1.31) or fibromyalgia related costs (0.85, 95% CI: 0.61 to 1.18) between treatments during the postindex period. CONCLUSION: No significant differences were found between pregabalin and duloxetine in the pre- to postindex change in mean all-cause or fibromyalgia-related total health care costs. PMID- 21615860 TI - Teeth fracture among visually impaired and sighted children of 12 and 15 years age groups of Udaipur city, India--a comparative study. AB - AIM: To compare the prevalence of anterior teeth fracture among sighted and visually impaired children of 12 and 15 years ages in Udaipur city, India. MATERIALS AND METHODS: A cross sectional study was carried out among 508 children (83 visually impaired and 425 sighted children), which comprised of 55 males and 28 females among visually impaired and 348 females and 77 males among sighted. The survey was carried out using Andreasen's classification for teeth fracture. Data was analyzed using chi-square test, with keeping level of significance at P < 0.05. RESULTS: It was found that, visually impaired children (32.5%) had significantly higher percentage of teeth fracture than that of sighted children (9.6%) (P = 001). Males had significantly higher percentage of teeth fracture than females in both groups (P = 0.001). It was observed that overjet of more than 3.5 mm had significantly increased risk of sustaining traumatic dental injuries among visually impaired (70.4%) than that of sighted individuals (46.3%) (P = 0.043). However, fracture of teeth was independent of age groups and lip coverage. CONCLUSION: Prevalence of traumatic dental injuries in a group of individuals with visual impairment was higher than that of sighted. This should alert caregivers to carry out a profound investigation of the events and suggest methods to reduce this type of morbidity. PMID- 21615862 TI - Considerations of maxillary tuberosity fractures during extraction of upper molars: a literature review. AB - BACKGROUND: Maxillary tuberosity fractures during molar teeth extraction commonly occur in dental practice; however, very few cases have been reported and discussed in the literature. A correct preoperative radiographic interpretation, coupled with the anatomical knowledge of the structures involved, is essential to prevent such complications. AIM: The purpose of this paper is to enumerate the predisposing and etiological factors of maxillary tuberosity fractures during the extraction of upper molars, discuss the procedures that need to be taken when small or large fractured fragments of the tuberosity are evident during surgery, and suggest appropriate recommendations. This study is based on a thorough literature review. CONCLUSIONS: Upon discovering that a maxillary tuberosity has fractured, the dentist must first halt the procedure before inadvertent laceration of the adjoining soft tissue occurs and then determine the extent of the fracture by palpating the mobile fragment. After performing the dissection of the soft tissues, immediate removal of the small fractures, including the tooth with small bony fragments, may be the best option, because of the difficulty incurred when attempting to retain the bone. When a large bony fragment is present, it is recommended (i) that the extraction be abandoned and surgical removal of the tooth be performed using root sectioning, (ii) that the dentist tries to detach the fractured tuberosity from the roots, or (iii) that the dentist stabilizes the mobile part(s) of the bone by means of a rigid fixation technique for 4-6 weeks and, at a future moment, attempts a surgical removal without the use of a forceps. PMID- 21615861 TI - Development of new artificial models for splint rigidity evaluation. AB - AIM: We developed two versions of an artificial model and assessed their suitability for splint rigidity evaluation. These models allowed the simulation of traumatically loosened teeth and the use of the acid-etch technique for splint application. MATERIALS AND METHODS: A straight and half-round arch bar model with bovine tooth facets were manufactured. Using the Periotest method, tooth mobility was evaluated before (PTVpre) and after (PTVpost) splinting. Two types of previously investigated wire-composite splints, WCS1 (Dentaflex 0.45 mm; Dentaurum) and WCS2 (Strengthens 0.8 * 1.8 mm; Dentaurum), were applied (n = 10) to each model. The relative splint effect (SpErel = DeltaPTV/PTVpre) was calculated, and the working times for the models and splints were evaluated. Student's t-test and the Mann-Whitney U-test were employed with Bonferroni correction for multiple hypotheses. RESULTS: When comparing the relative splint effect of the 'injured' central incisors between the models within one splint type, differences were only found for tooth 21 (WCS2; P < 0.008); for comparisons of splints within one model type, differences were detected for both incisors and model types (P < 0.008). With the straight model, significantly less working time was necessary (P < 0.05). CONCLUSION: Using these models for in vitro splint rigidity evaluation, the splints can be applied with the acid-etch technique and tooth mobility can be individually adjusted. WCS1 is considered flexible compared to the more rigid WCS2. The results from the straight and the round model were predominantly closely related to each other. In terms of working time, the straight model is superior to the round model. PMID- 21615863 TI - Treatment of oblique crown fractures in maxillary premolars using adhesive tooth fragment reattachment: 19 years of follow up. AB - This report presents an oblique crown fracture in the maxillary right premolars of an adolescent because of fall, which was treated using adhesive tooth fragment reattachment. The impact of the mandible base to the floor caused minor fractures in multiple teeth, severe fracture of teeth 14 and 15, and condylar fractures. The fragments of teeth 43, 45, 46, and 16 were lost at the site of accident. The condylar fractures were managed with a non-surgical conservative approach combining mandibular immobilization and stabilization of temporomandibular joints with maxillomandibular fixation using acrylic splints. After 3 months, the interocclusal splints were removed, and the patient was referred for dental care. Teeth 14 and 15 presented complicated oblique crown fractures causing separation of the buccal and palatal fragments, which were in place, attached to the gingival tissue. Tooth 15 presented chronic hyperplastic pulpits. The fragments were banded for stabilization during the endodontic treatment, and the bands served as matrix for adhesive tooth fragment reattachment. The other fractured teeth received direct composite resin restorations. After 8 years, tooth 16 developed pulp necrosis and was treated endodontically and restored with composite resin. Clinical and radiographic examination 19 years after trauma showed a good adaptation of the tooth fragment/composite resin restoration, good periodontal health, no signs of root resorption, and intact lamina dura. PMID- 21615864 TI - Clinical management of a fractured anterior tooth with reattachment technique: a case report with an 8-year follow up. AB - Fracture of anterior teeth is a relatively common outcome of trauma to the teeth. If the fractured teeth fragments are recovered by the patient and brought to the dental office within reasonable time, the fragments may be reattached to the remaining tooth structure. This article presents a case of a 15-year-old male patient with fractured left maxillary lateral incisor. The fragment reattachment was performed using dual-cured composite resin cement and the treatment outcomes of an 8-year follow up were presented. The technique described in this case report for reattachment of the fractured fragments is simple, while restoring providing long-lasting esthetics and improved function with a very conservative approach. PMID- 21615866 TI - A randomized, controlled study comparing the cosmetic outcome of a new wound closure device with Prolene suture closing caesarean wounds. AB - A prospective, randomised study was conducted to compare the wound closure performance and cosmetic outcome of caesarean section wounds closed with traditional Prolene suture or a new wound closure device (Leukosan((r)) SkinLink). Sixty-one patients referred to primary section were allocated to wound closure with either Leukosan((r)) SkinLink or Prolene suture. Cosmetic outcome as the primary measure was evaluated by the patient, the surgeon as well as by independent examiners blinded to the method of wound closure. Evaluations were recorded at 3, 6 and 12 months following wound closure. Both methods of wound closure scored equally high on the visual analogue scale for cosmetic evaluation at the 3-, 6- and 12-month follow-ups as assessed by the patient, surgeon and the blinded observers. The study has shown that innovative methods for wound closure compared with traditional methods such as suture providing excellent cosmetic results represent a valid alternative to physician and patient for surgical incisions. PMID- 21615865 TI - Breastfeeding and long-chain polyunsaturated fatty acid intake in the first 4 post-natal months and infant cognitive development: an observational study. AB - The aim of this study was to examine infant feeding and the long-chain polyunsaturated fatty acid (LCPUFA) concentration of breast milk and formulas in relation to infant development. The prospective Pregnancy, Infection and Nutrition Study (n=358) collected data on breastfeeding, breast milk samples and the formulas fed through 4months post-partum. At 12months of age, infants' development was assessed (Mullen Scales of Early Learning). Linear regression was used to examine development in relation to breastfeeding, breast milk docosahexaenoic acid (DHA) and arachidonic acid (AA) concentration, and DHA and AA concentration from the combination of breast milk and formula. The median breast milk DHA concentration was 0.20% of total fatty acids [interquartile range (IQR)=0.14, 0.34]; median AA concentration was 0.52% (IQR=0.44, 0.63). Upon adjustment for preterm birth, sex, smoking, race and ethnicity and education, breastfeeding exclusivity was unrelated to development. Among infants exclusively breastfed, breast milk LCPUFA concentration was not associated with development (Mullen composite, DHA: adjusted beta=-1.3, 95% confidence interval: -10.3, 7.7). Variables combining DHA and AA concentrations from breast milk and formula, weighted by their contribution to diet, were unassociated with development. We found no evidence of enhanced infant development related to the LCPUFA content of breast milk or formula consumed during the first four post-natal months. PMID- 21615867 TI - Cisapride: what can we learn from the rise and fall of a prokinetic? AB - Cisapride, the prototype serotonergic agent, evolved from a body of research that defined the key roles of serotonergic receptors in gastrointestinal motor and sensory function. Impressed by its in vitro properties and encouraged by clinical trial data, cisapride became the drug of choice for the treatment of a wide range of motility disorders and clinicians appeared impressed by its efficacy and comfortable with its side-effect profile. Once serious cardiac events began to be reported in association with cisapride therapy, dark clouds rapidly gathered and soon enveloped the drug, leading to its widespread withdrawal from markets. What lessons can we learn from the story of cisapride? How can its brief but spectacular rise and equally sensational demise inform the development of new drugs which are so sorely needed in the management of motility and functional gastrointestinal disorders? This review explores the background to the development of cisapride, its history in clinical trials and the experience with adverse events and, in so doing, attempts to identify lessons for the future in the therapeutics of enteric neuromodulatory drugs. PMID- 21615868 TI - Nutrigenomic approach to tackle the unpleasant journey to Helicobacter pylori associated gastric carcinogenesis. AB - While dietary habits or nutritional intake continue to rank as significant factors influencing the incidence of cancer, there have been considerable scientific uncertainties about who will benefit, but who about will not benefit from nutrition. This might be due to inadequate knowledge about an individual's genetic background, the cumulative effect of nutrients on genetic expression profiles, ambiguous clinical differences between beneficiaries and non beneficiaries and the lack of information about active protein induction. During the past 200 years of nutrition research, we have experienced revolutionary advances in both chemistry and genomics. According to the high expectations for tailored medicine, a nutrigenomic approach harboring tremendous potential to change the future of dietary guideline and personal recommendations will provide an essential basis for personalized dietary recommendations to prevent common multifactorial diseases decades before their overt clinical manifestation. In the current review, we introduce our efforts to discover Helicobacter pylori (H. pylori)-related disease biomarkers applicable for diagnostic, predictive and therapeutic purposes using several kinds of technology. For instance, based on publications showing the in vitro and in vivo efficacy of Korean red ginseng on mitigating H. pylori-associated gastric atrophy, a nutrigenomic approach allows us to confirm that Korean red ginseng prevents H. pylori-associated gastric cancer in predictable ways. PMID- 21615869 TI - Efficacy and safety of adalimumab in Crohn's disease: meta-analysis of placebo controlled trials. AB - OBJECTIVE: To evaluate the efficacy and safety of adalimumab in inducing and maintaining remission of Crohn's disease. METHODS: Electronic databases were searched. Placebo-controlled trials of adalimumab used in patients with Crohn's disease were included. Data were analyzed with Review Manager 4.2. RESULTS: Four studies enrolling 1402 patients were confirmed as meeting our criteria. Remission rates of inducing and maintaining remission in patients with Crohn's disease were higher for adalimumab than placebo (P<0.05). Adalimumab significantly improves the quality of life in patients with Crohn's disease. No significant difference in total adverse events was found in maintaining remission. Moreover, there were significantly less serious adverse events from taking adalimumab than from taking the placebo in long-term treatment. Patients with increased baseline C-reactive protein may benefit more from adalimumab therapy than those without. CONCLUSIONS: Adalimumab is effective and safe in Crohn's disease. However, studies of a larger number of patients are still required for better assessing the safety profile of adalimumab. PMID- 21615870 TI - Gastroesophageal and laryngopharyngeal reflux detected by 24-hour combined impedance and pH monitoring in healthy Chinese volunteers. AB - OBJECTIVE: To define the normative data of gastroesophageal reflux (GER) and laryngopharyngeal reflux (LPR) in healthy Chinese volunteers. METHODS: Healthy volunteers without gastrointestinal and throat disease and symptoms were enrolled and underwent 24-hour impedance-pH monitoring. RESULTS: A total of 37 healthy volunteers completed all the tests (female 54.1%; age 38.0+/-15.2 years). Of 1862 GER, 49.6% were mixed, 40.9% were liquid and 9.5% were gas. More than half (54.9%) were acid. The median and 95th percentile numbers of GER were 52 and 71, respectively. The number of LPR was eight, of which two were mixed and the others were liquid. Only one episode of reflux was classified as acid LPR. The median and 95th percentile numbers of LPR were 0 and 2, respectively. More liquid reflux were associated with LPR (P=0.02). Age had no influence on GER and LPR reflux profiles. CONCLUSIONS: Few LPR occurred in healthy Chinese volunteers and they were rarely acid when they reached the larynx. LPR was mostly associated with liquid reflux. PMID- 21615871 TI - Chemopreventive effect of epigallocatechin-3-gallate (EGCG) and folic acid on the N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced gastrointestinal cancer in rat model. AB - OBJECTIVE: To investigate the chemopreventive effect and mechanisms of epigallocatechin-3-gallate (EGCG) and folic acid on N-methyl-N'-nitro-N nitrosoguanidine (MNNG)-induced gastrointestinal cancer in rats, and to investigate and compare the combinatorial effects of EGCG and folic acid on the chemoprevention of gastrointestinal carcinogenesis. METHODS: A total of 159 healthy male Wistar rats were randomly divided into seven groups to have the MNNG in drink (group M), MNNG in drink and EGCG in the feed (group ME), MNNG in drink and folic acid in the feed (group MF), MNNG in drink and EGCG+folic acid in the feed (group MEF), EGCG in the feed (group E), folic acid in the feed (group F) or normal feed (group C), respectively. At 44 weeks, all the rats were killed and assessed for the presence of gastrointestinal tumor. The occurrence of cancer was evaluated by histology. Ki-67 in cancerous tissues and in situ apoptosis were determined by immunohistochemical staining or terminal deoxyribonucleotide transferase-mediated nick-end labeling (TUNEL) assay, respectively. RESULTS: The experiment was completed in 157 rats (98.74%). As compared with group M, the tumor incidence of group MEF decreased significantly (P=0.011). Ki-67 expression in cancerous tissues of group ME and MEF also decreased significantly (P=0.038, P=0.009), while apoptosis of group ME, MF and MEF increased significantly (P=0.000, P=0.003, P=0.000). CONCLUSION: EGCG combined with folic acid has an obvious chemopreventive effect on gastrointestinal carcinogenesis induced by MNNG in rats. PMID- 21615872 TI - Diagnostic value of terminal ileum biopsy in chronic diarrhea with normal endoscopic appearance. AB - OBJECTIVE: To investigate the diagnostic value of a terminal ileum biopsy in chronic non-bloody diarrhea with normal endoscopic appearance. METHODS: Patients who had a terminal ileum biopsy performed between January 2007 and January 2010 during a colonoscopy despite normal endoscopic appearance were reviewed. They were divided into two groups according to the indication for the colonoscopy: non bloody diarrhea and non-diarrhea; and their histopathological findings were compared. RESULTS: By screening the colonoscopy records, 473 patients were found to have had a terminal ileum biopsy. Of these patients, 178 were excluded for various reasons. Finally, 295 patients were suitable for analysis. The non-bloody diarrhea group consisted of 135 patients, while the remaining 160 patients constituted the non-diarrhea group. A histopathological examination of the terminal ileum biopsy specimens in the non-bloody diarrhea group revealed an abnormal histology in eight patients (5.9%), compared with five patients (3.1%) in the non-diarrhea group. No statistically significant difference was found between the two groups (P=0.26). CONCLUSION: Although performing a biopsy of the terminal ileum despite a macroscopically normal appearance may offer some additional benefit to the diagnosis of chronic non-bloody diarrhea, prospective trials are needed for assessing the diagnostic value of a terminal ileum biopsy in patients with chronic diarrhea. PMID- 21615873 TI - Clinical features and molecular alterations of traditional serrated adenoma in sporadic colorectal carcinogenesis. AB - OBJECTIVE: To compare clinical features and molecular alterations between traditional serrated adenomas (TSA) & serrated carcinomas (SCa) and traditional adenomas (TA) & carcinomas (Ca) of the colorectum and to verify a traditional serrated pathway of sporadic colorectal carcinogenesis. METHODS: One thousand two hundred slides of colorectal polyps obtained from 1160 patients were collected and reviewed to define TSA and clinical and pathological features were analyzed and compared with TA. DNA was extracted from specimens of TSA, TA, SCa and Ca, v raf murine sarcoma viral oncogene homolog B (BRAF) V600E mutation and microsatellite instability (MSI) (assay for BAT25 and BAT26) were analyzed. RESULTS: Overall 29 TSA were confirmed (2.5%), and there was an age difference between patients with TSA and TA (56.0 vs 62.7, P<0.05). Compared with TA, TSA was located more often in the rectosigmoid colon (TSA 62.1% vs TA 35.2%, P<0.05), but occurred less in the descending colon (TSA 0% vs TA 25.35%, P=0.0068). No difference was found in terms of gender and the size or pedicles of polyps (P>0.05). The BRAF V600E mutation was detected in 36.3% of SCa and 26.7% of TSA patients, but it was not detected in TA and Ca patients; MSI-H was noticed in 23% of SCa, 33.3% of TSA, 5.3% of Ca and 0% of TA patients, respectively (P<0.05). CONCLUSION: There might be a traditional serrated pathway of sporadic colorectal carcinogenesis that is different from the conventional adenoma to carcinoma carcinogenesis pathway in the colorectum. PMID- 21615874 TI - Evaluation of three different hepatitis C virus typing methods for detection of mixed-genotype infections. AB - OBJECTIVE: To evaluate the clinical applicability of an eligible assay for the true prevalence of hepatitis C virus (HCV) mixed-genotype infections. METHODS: A newly developed HCV genotyping method targeting all six major HCV genotypes and 12 subtypes, restriction fragment length polymorphism (RFLP) and a serotyping assay were utilized for the detection of HCV mixed-genotype infections using known HCV genotypes and unknown samples. RESULTS: In a defined mix of HCV genotypes, a genotype present at levels as low as 8.3% was detected by our newly developed assay, showing a threefold increase in sensitivity over that of direct deoxyribonucleic (DNA) sequencing. A comparative study of the accuracy among the three genotyping methods was carried out on samples obtained from 50 thalassemic patients who received multiple blood transfusions. The results showed that viruses in approximately 42% of the samples from this group were determined to be infected with mixed genotypes by our newly developed method. A serotyping assay and RFLP analysis, performed with poor results, could identify only 18% and 10% of mixed-genotype infections, respectively. CONCLUSION: The newly developed assay may be the method of choice when detection of genotypes present at low levels in mixed-genotype infections due to its higher level of sensitivity. PMID- 21615875 TI - Low-dose hepatic computed tomography perfusion imaging and its preliminary study. AB - OBJECTIVE: Computed tomography perfusion imaging (CTPI) is a rapid and non invasive functional imaging method that reflects hemodynamic changes of liver diseases. However, its large radiation dosage limits its clinical application. We aimed to evaluate the feasibility of low-dose CTPI in normal liver and its preliminary application in hepatocellular carcinoma (HCC). METHODS: CTPI was performed in 34 healthy volunteers randomly divided into three groups with different applications of tube current, including a conventional dose group, a median-dose group and a low-dose group. The perfusion parameters of each group were compared and a low-dose CTPI was performed in 13 patients with HCC. RESULTS: Relatively satisfying images and perfusion parameters of liver CTPI were acquired with the different tube currents. There were no significant differences between the parameters of the three groups (P>0.05). The effective dosage of conventional, median and low-dose liver CTPI were 19.62 mSv, 12.61 mSv, and 7.01 mSv, respectively. The radiation dosage of low-dose liver CTPI was reduced to 64.27% compared with that of the conventional group. The hepatic blood flow, hepatic blood volume and hepatic perfusion index of HCC were higher than background liver parenchyma and normal liver. CONCLUSIONS: Low-dose liver CTPI obtained similar perfusion parameters result to that of the conventional-dose, whereas the radiation dosage was reduced by 2/3. Low-dose liver CTPI can reflect the hemodynamic change of HCC. Low-dose liver CTPI has potential clinical value for diagnosis and differential diagnosis of liver diseases. PMID- 21615876 TI - Characteristics of pancreatic diabetes in patients with autoimmune pancreatitis. AB - OBJECTIVE: Although patients with autoimmune pancreatitis (AIP) tend to have concurrent diverse disorders, very few studies have focused on diabetes mellitus (DM) coexisting with AIP. METHODS: In total 102 AIP patients with DM were divided into three groups. Those with DM before the onset of AIP were labeled group A (n=35), those who developed DM and AIP simultaneously were labeled group B (n=58) and those who developed DM after steroid therapy for AIP were labeled group C (n=9). The characteristics of DM among the three groups were evaluated. RESULTS: No significant differences were noted in the age of DM onset among the three groups. However, the mean duration of DM was significantly longer in group A (8.7 years) than in groups B and C. AIP developed 6.8 years after DM onset in group A, whereas it developed 1.8 years after steroid therapy in group C. Group A had the highest rate (25.7%) of family members with a history of AIP. Levels of serum albumin, total cholesterol and triglyceride were significantly lower in group A. No correlations were found between glycated hemoglobin and benzoyl-tyrosyl para aminobenzoic acid. Hypoglycemia was observed in 20% of patients under insulin therapy. Most of them were habitual drinkers and received no pancreatic enzymes. Group A showed a high prevalence of retinopathy, nephropathy and macrovascular disorders than group B. CONCLUSION: Aspects of AIP-associated pancreatic diabetes were clarified. AIP-associated DM must be controlled by a full assessment of the pancreatic endocrine and exocrine function. PMID- 21615877 TI - An experimental study of betadine irrigation for preventing infection during the natural orifice transluminal endoscopic surgery (NOTES) procedure. AB - OBJECTIVE: To study the effect and feasibility of using betadine irrigation of the gastrointestinal tract for preventing infection during the natural orifice transluminal endoscopic surgery (NOTES) procedure. METHODS: Twelve sows were used in this study. Four sows in the control group were lavaged with 500 mL saline. The eight sows in the experimental group were first lavaged with 500 mL saline and then irrigated with 200 mL betadine. A total of 5 mL of gastrointestinal (GI) tract fluid was collected before and after lavage, respectively, and 5 mL of peritoneal fluid was collected at the end of the NOTES procedure. A follow-up endoscopic examination of the GI tract was performed 24 h after NOTES. The animals were killed and necropsied after 3 weeks. RESULTS: Irrigation with betadine of the GI tract significantly reduced the bacterial load of GI fluid. One sow died of diaphragmatic injury. No inflammation, ulcer or bleeding were observed in the experimental group by endoscopy after 24 h. More adhesions and abscesses were found in the control group than in the experimental group after 3 weeks. Only one case of adhesion was observed in the experimental group using the transcolonic approach. CONCLUSIONS: Betadine irrigation of the GI tract is effective and feasible for preventing infection during the NOTES procedure. Further studies are needed for assessing the effectiveness and safety of betadine irrigation in the clinical application of NOTES. PMID- 21615878 TI - Magnetic resonance cholangiography to evaluate biliary tree integrity after cholecystectomy: a case report. PMID- 21615879 TI - Synchronous multifocal early gastric cancers and rectal adenocarcinoma: lesson learnt from image-enhanced endoscopy. PMID- 21615880 TI - Glutathione S-transferase M1/T1 gene polymorphisms and vitiligo in a Mediterranean population. PMID- 21615881 TI - Clinical outcome and pathological features associated with NRAS mutation in cutaneous melanoma. AB - The effect of NRAS mutations on the pathological features and clinical outcomes in patients with cutaneous melanoma was compared with that of tumors containing BRAF(V600E) mutations and tumors wild type for both (WT). Clinical outcome data were obtained from a prospective cohort of 249 patients. Mutations involving NRAS and BRAF(V600E) were detected by PCR and were sequence verified. Cox proportional hazards regression was performed to relate NRAS and BRAF mutations to clinical outcome. Seventy-five percentage of NRAS mutations occurred in tumors >1 mm thick (BRAF(V600E) 40%, WT 34%); 75% of NRAS mutations had >1 mitosis/mm(2) (BRAF(V600E) 40%, WT 55%). When compared to WT, multivariate analysis of melanoma specific survival (MSS) identified NRAS mutations as an adverse prognostic factor [hazard ratio (HR) 2.96; P = 0.04] but not BRAF(V600E) mutations (HR 1.73; P = 0.23). NRAS mutations were associated with thicker tumors and higher rates of mitosis when compared to BRAF(V600E) and WT melanoma and independently of this, with shorter MSS. PMID- 21615882 TI - Treating major depression by creating positive expectations for the future: a pilot study for the effectiveness of future-directed therapy (FDT) on symptom severity and quality of life. AB - INTRODUCTION: This nonrandomized pilot study assesses the efficacy of a new future-oriented form of therapy, known as future-directed therapy (FDT), as a treatment for patients with Major Depressive Disorder (MDD) in a naturalistic hospital-based outpatient psychiatry clinic. The study measured symptom severity of depression and anxiety, in addition to quality of life pre- and posttreatment. AIMS: The study examined a new manualized treatment designed to help people anticipate a more positive future. The intervention consists of twenty 90-min group sessions administered twice a week over 10 weeks. The intervention was compared to depressed patients in the same clinic who enrolled in traditional cognitive-based group psychotherapy. Sixteen patients with MDD completed the FDT intervention as part of their outpatient treatment for depression. Seventeen patients with MDD participated in treatment as usual (TAU) cognitive-based group therapy. The Quick Inventory of Depressive Symptoms, the Beck Anxiety Inventory, and the Quality-of-Life Enjoyment and Satisfaction Questionnaire short form, self report instruments were administered prior to and immediately after the completion of therapy. RESULTS: Patients treated with FDT demonstrated significant improvements in depression (P = 0.001), anxiety (P = 0.021) and quality of life (P = 0.035), and also reported high satisfaction with the therapy. Compared to the TAU group, patients treated with FDT showed greater improvements in depressive symptoms (P = 0.049). CONCLUSIONS: FDT may have the potential of becoming an additional treatment option for patients with MDD. PMID- 21615883 TI - Associations of body mass index and waist circumference with: energy intake and percentage energy from macronutrients, in a cohort of Australian children. AB - BACKGROUND: It is evident from previous research that the role of dietary composition in relation to the development of childhood obesity remains inconclusive. Several studies investigating the relationship between body mass index (BMI), waist circumference (WC) and/or skin fold measurements with energy intake have suggested that the macronutrient composition of the diet (protein, carbohydrate, fat) may play an important contributing role to obesity in childhood as it does in adults. This study investigated the possible relationship between BMI and WC with energy intake and percentage energy intake from macronutrients in Australian children and adolescents. METHODS: Height, weight and WC measurements, along with 24 h food and drink records (FDR) intake data were collected from 2460 boys and girls aged 5-17 years living in the state of Queensland, Australia. RESULTS: Statistically significant, yet weak correlations between BMI z-score and WC with total energy intake were observed in grades 1, 5 and 10, with only 55% of subjects having a physiologically plausible 24 hr FDR. Using Pearson correlations to examine the relationship between BMI and WC with energy intake and percentage macronutrient intake, no significant correlations were observed between BMI z-score or WC and percentage energy intake from protein, carbohydrate or fat. One way ANOVAs showed that although those with a higher BMI z-score or WC consumed significantly more energy than their lean counterparts. CONCLUSION: No evidence of an association between percentage macronutrient intake and BMI or WC was found. Evidently, more robust longitudinal studies are needed to elucidate the relationship linking obesity and dietary intake. PMID- 21615884 TI - MMP28 (epilysin) as a novel promoter of invasion and metastasis in gastric cancer. AB - BACKGROUND: The purpose of this study was to investigate invasion and metastasis related genes in gastric cancer. METHODS: The transwell migration assay was used to select a highly invasive sub-line from minimally invasive parent gastric cancer cells, and gene expression was compared using a microarray. MMP28 upregulation was confirmed using qRT-PCR. MMP28 immunohistochemistry was performed in normal and gastric cancer specimens. Invasiveness and tumor formation of stable cells overexpressing MMP28 were tested in vitro and in vivo. RESULTS: MMP28 was overexpressed in the highly invasive sub-cell line. Immunohistochemistry revealed MMP28 expression was markedly increased in gastric carcinoma relative to normal epithelia, and was significantly associated with depth of tumor invasion, lymph node metastasis and poorer overall survival. Ectopic expression of MMP28 indicated MMP28 promoted tumor cell invasion in vitro and increased gastric carcinoma metastasis in vivo. CONCLUSIONS: This study indicates MMP28 is frequently overexpressed during progression of gastric carcinoma, and contributes to tumor cell invasion and metastasis. MMP28 may be a novel therapeutic target for prevention and treatment of metastases in gastric cancer. PMID- 21615885 TI - Multiple congenital ocular anomalies in Icelandic horses. AB - BACKGROUND: Multiple congenital ocular anomalies (MCOA) syndrome is a hereditary congenital eye defect that was first described in Silver colored Rocky Mountain horses. The mutation causing this disease is located within a defined chromosomal interval, which also contains the gene and mutation that is associated with the Silver coat color (PMEL17, exon 11). Horses that are homozygous for the disease causing allele have multiple defects (MCOA-phenotype), whilst the heterozygous horses predominantly have cysts of the iris, ciliary body or retina (Cyst phenotype). It has been argued that these ocular defects are caused by a recent mutation that is restricted to horses that are related to the Rocky Mountain Horse breed. For that reason we have examined another horse breed, the Icelandic horse, which is historically quite divergent from Rocky Mountain horses. RESULTS: We examined 24 Icelandic horses and established that the MCOA syndrome is present in this breed. Four of these horses were categorised as having the MCOA-phenotype and were genotyped as being homozygous for the PMEL17 mutation. The most common clinical signs included megaloglobus, iris stromal hypoplasia, abnormal pectinate ligaments, iridociliary cysts occasionally extending into the peripheral retina and cataracts. The cysts and pectinate ligament abnormalities were observed in the temporal quadrant of the eyes. Fourteen horses were heterozygous for the PMEL17 mutation and were characterized as having the Cyst-phenotype with cysts and occasionally curvilinear streaks in the peripheral retina. Three additional horses were genotyped as PMEL17 heterozygotes, but in these horses we were unable to detect cysts or other forms of anomalies.One eye of a severely vision-impaired 18 month-old stallion, homozygous for the PMEL17 mutation was examined by light microscopy. Redundant duplication of non-pigmented ciliary body epithelium, sometimes forming cysts bulging into the posterior chamber and localized areas of atrophy in the peripheral retina were seen. CONCLUSIONS: The MCOA syndrome is segregating with the PMEL17 mutation in the Icelandic Horse population. This needs to be taken into consideration in breeding decisions and highlights the fact that MCOA syndrome is present in a breed that are more ancient and not closely related to the Rocky Mountain Horse breed. PMID- 21615886 TI - Cardiac safety of indacaterol in healthy subjects: a randomized, multidose, placebo- and positive-controlled, parallel-group thorough QT study. AB - BACKGROUND: Indacaterol is a novel once-daily ultra long-acting beta2-agonist for the treatment of chronic obstructive pulmonary disease. It is known that beta2 agonists, like other adrenergic compounds, can prolong the QT-interval. This thorough QT/QTc study (as per ICH E14 guideline) evaluated the effect of indacaterol on the QT interval in healthy subjects. METHODS: In this randomized, double-blind, parallel-group, placebo- and positive-controlled (open-label moxifloxacin) study, non-smoking healthy subjects (18-55 years, body mass index: 18.5-32.0 kg/m2) were randomized (4:4:2:4:1) to 14-day treatment with once-daily indacaterol (150 MUg, 300 MUg, or 600 MUg), placebo, or placebo/moxifloxacin (double-blind 14-day treatment with placebo and a single open-label dose of 400 mg moxifloxacin on Day 14). The primary endpoint was the change from baseline on Day 14 in QTcF (QT interval corrected for heart rate using Fridericia's formula). RESULTS: In total, 404 subjects were randomized to receive indacaterol (150 [n = 108], 300 [n = 108], 600 MUg [n = 54]), placebo (n = 107), or placebo/moxifloxacin (n = 27); 388 subjects completed the study. Maximal time matched mean (90% confidence intervals) treatment differences from placebo in QTcF change from baseline on Day 14 were 2.66 (0.55, 4.77), 2.98 (1.02, 4.93) and 3.34 (0.86, 5.82) ms for indacaterol 150 MUg, 300 MUg and 600 MUg, respectively. Study sensitivity was confirmed with moxifloxacin demonstrating a significant maximal time-matched QTcF prolongation of 13.90 (10.58, 17.22) ms compared to placebo. All indacaterol doses were well tolerated. CONCLUSION: Indacaterol, at doses up to 600 MUg once daily (2-4 times the therapeutic dose) does not have any clinically relevant effect on the QT interval. PMID- 21615887 TI - Toxoplasma, toxocara and tuberculosis co-infection in a four year old child. AB - BACKGROUND: Tuberculosis. toxocariasis and toxoplasmosis are among the common infectious causes of lymphadenitis in children. Cases of Toxoplasma gondii and Toxocara spp co-infection have been reported. CASE PRESENTATION: This case report describes a co-infection of Toxoplasma gondii, Toxocara spp and tuberculosis in a child with chronic lymphadenopathy and eosinophilia. CONCLUSION: The case report highlights two important points. First is the diagnostic challenges that are encountered by clinicians in tropical countries such as Sri Lanka, where lymphadenopathy and eosinophilia with a positive serology commonly point towards a parasitic infection. Secondly the importance of proper history taking and performing the Mantoux test as a first line investigation in a country where the incidence of tuberculosis is low, even in the absence of a positive contact history. PMID- 21615888 TI - Neurocysticercosis as an important differential of seizures in pregnancy: two case reports. AB - INTRODUCTION: Seizures in pregnancy usually result from eclampsia, epilepsy or central nervous system disorders. Neurocysticercosis is a rare, but an important, cause of first-time convulsions in pregnancy. CASE PRESENTATIONS: We report the cases of two Indian women, aged 20 and 24 years old respectively, with neurocysticercosis presenting in the second trimester of pregnancy with convulsions. Both had marginally raised blood pressure with 1+ urine albumin and neither had a past history of convulsions. The neurocysticercosis was diagnosed on magnetic resonance imaging of the head, which showed spherical ring-enhancing lesions in the brain. In one woman, pregnancy was terminated due to spina bifida in the fetus and she was discharged on albendazole and phenytoin. The second woman was put on carbamazepine: she had an emergency Cesarean section at term for fetal distress and delivered a healthy baby. Her postnatal period was uneventful. CONCLUSION: Neurocysticercosis should be considered in pregnant women presenting with seizures which cannot be explained by eclampsia, especially in early pregnancy. PMID- 21615889 TI - Multiple testing for gene sets from microarray experiments. AB - BACKGROUND: A key objective in many microarray association studies is the identification of individual genes associated with clinical outcome. It is often of additional interest to identify sets of genes, known a priori to have similar biologic function, associated with the outcome. RESULTS: In this paper, we propose a general permutation-based framework for gene set testing that controls the false discovery rate (FDR) while accounting for the dependency among the genes within and across each gene set. The application of the proposed method is demonstrated using three public microarray data sets. The performance of our proposed method is contrasted to two other existing Gene Set Enrichment Analysis (GSEA) and Gene Set Analysis (GSA) methods. CONCLUSIONS: Our simulations show that the proposed method controls the FDR at the desired level. Through simulations and case studies, we observe that our method performs better than GSEA and GSA, especially when the number of prognostic gene sets is large. PMID- 21615890 TI - Susceptibility of adult female Aedes aegypti (Diptera: Culicidae) to the entomopathogenic fungus Metarhizium anisopliae is modified following blood feeding. AB - BACKGROUND: The mosquito Aedes aegypti, vector of dengue fever, is a target for control by entomopathogenic fungi. Recent studies by our group have shown the susceptibility of adult A. aegypti to fungal infection by Metarhizium anisopliae. This fungus is currently being tested under field conditions. However, it is unknown whether blood-fed A. aegypti females are equally susceptible to infection by entomopathogenic fungi as sucrose fed females. Insect populations will be composed of females in a range of nutritional states. The fungus should be equally efficient at reducing survival of insects that rest on fungus impregnated surfaces following a blood meal as those coming into contact with fungi before host feeding. This could be an important factor when considering the behavior of A. aegypti females that can blood feed on multiple hosts over a short time period. METHODS: Female A. aegypti of the Rockefeller strain and a wild strain were infected with two isolates of the entomopathogenic fungus M. anisopliae (LPP 133 and ESALQ 818) using an indirect contact bioassay at different times following blood feeding. Survival rates were monitored on a daily basis and one way analysis of variance combined with Duncan's post-hoc test or Log-rank survival curve analysis were used for statistical comparisons of susceptibility to infection. RESULTS: Blood feeding rapidly reduced susceptibility to infection, determined by the difference in survival rates and survival curves, when females were exposed to either of the two M. anisopliae isolates. Following a time lag which probably coincided with digestion of the blood meal (96-120 h post feeding), host susceptibility to infection returned to pre-blood fed (sucrose fed) levels. CONCLUSIONS: Reduced susceptibility of A. aegypti to fungi following a blood meal is of concern. Furthermore, engorged females seeking out intra domicile resting places post-blood feeding, would be predicted to rest for prolonged periods on fungus impregnated black cloths, thus optimizing infection rates. It should be remembered that lowered susceptibility was only a temporary phenomenon and this may not necessarily occur when mosquitoes are infected with other fungal isolates. These results may have implications for field testing of entomopathogenic fungi by our group and further studies should be carried out to better understand the insect-fungus interaction. PMID- 21615891 TI - Withdrawal-induced delirium associated with a benzodiazepine switch: a case report. AB - INTRODUCTION: Introduced in the early 1960s, diazepam remains among the most frequently prescribed benzodiazepine-type sedatives and hypnotics. Patients with chronic use of short-acting benzodiazepines are frequently switched to diazepam because the accumulating, long-acting metabolite, N-desmethyl-diazepam, prevents benzodiazepine-associated withdrawal symptoms, which can occur during trough plasma levels of short-acting benzodiazepines. Although mild to moderate withdrawal symptoms are frequently observed during benzodiazepine switching to diazepam, severe medical complications associated with this treatment approach have thus far not been reported. CASE PRESENTATION: A 64-year-old female Caucasian with major depression, alcohol dependence and benzodiazepine dependence was successfully treated for depression and, after lorazepam-assisted alcohol detoxification, was switched from lorazepam to diazepam to facilitate benzodiazepine discontinuation. Subsequent to the benzodiazepine switch, our patient unexpectedly developed an acute delirious state, which quickly remitted after re-administration of lorazepam. A newly diagnosed early form of mixed dementia, combining both vascular and Alzheimer-type lesions, was found as a likely contributing factor for the observed vulnerability to benzodiazepine induced withdrawal symptoms. CONCLUSION: Chronic use of benzodiazepines is common in the elderly and a switch to diazepam often precedes benzodiazepine discontinuation trials. However, contrary to common clinical practice, benzodiazepine switching to diazepam may require cross-titration with slow tapering of the first benzodiazepine to allow for the build-up of N-desmethyl diazepam, in order to safely prevent severe withdrawal symptoms. Alternatively, long-term treatment with low doses of benzodiazepines may be considered, especially in elderly patients with chronic use of benzodiazepines and proven vulnerability to benzodiazepine-associated withdrawal symptoms. PMID- 21615892 TI - Discriminative ability of the generic and condition-specific Child-Oral Impacts on Daily Performances (Child-OIDP) by the Limpopo-Arusha School Health (LASH) project: a cross-sectional study. AB - BACKGROUND: Generic and condition-specific (CS) oral-health-related quality-of life (OHRQoL) instruments assess the impacts of general oral conditions and specific oral diseases. Focusing schoolchildren from Arusha and Dar es Salaam, in Tanzania, this study compared the discriminative ability of the generic Child OIDP with respect to dental caries and periodontal problems across the study sites. Secondly, the discriminative ability of the generic-and the CS Child OIDP attributed to dental caries, periodontal problems and malocclusion was compared with respect to various oral conditions as part of a construct validation. METHODS: In Arusha, 1077 school children (mean age 14.9 years, range 12-17 years) and 1601 school children in Dar es Salaam (mean age 13.0 years, range 12-14 years) underwent oral clinical examinations and completed the Kiswahili version of the generic and CS Child-OIDP inventories. The discriminative ability was assessed as differences in overall mean and prevalence scores between groups, corresponding effect sizes and odd ratios, OR. RESULTS: The differences in the prevalence scores and the overall mean generic Child-OIDP scores were significant between the groups with (DMFT > 0) and without (DMFT = 0) caries experience and with (simplified oral hygiene index [OHI-S] > 1) and without periodontal problems (OHI-S <= 1) in Arusha and Dar es Salaam. In Dar es Salaam, differences in the generic and CS Child-OIDP scores were observed between the groups with and without dental caries, differences in the generic Child-OIDP scores were observed between the groups with and without periodontal problems, and differences in the CS Child-OIDP scores were observed between malocclusion groups. The adjusted OR for the association between dental caries and the CS Child-OIDP score attributed to dental caries was 5.4. The adjusted OR for the association between malocclusion and CS Child-OIDP attributed to malocclusion varied from 8.8 to 2.5. CONCLUSION: The generic Child-OIDP discriminated equally well between children with and without dental caries and periodontal problems across socio-culturally different study sites. Compared with its generic form, the CS Child-OIDP discriminated most strongly between children with and without dental caries and malocclusion. The CS Child OIDP attributed to dental caries and malocclusion seems to be better suited to support clinical indicators when estimating oral health needs among school children in Tanzania. PMID- 21615893 TI - Half-dose verteporfin photodynamic therapy for bullous variant of central serous chorioretinopathy: a case report. AB - INTRODUCTION: Central serous chorioretinopathy is characterized by serous neurosensory detachment of the macula and it usually resolves spontaneously with good visual prognosis. In some patients, however, the serous retinal detachment might be very extensive and can result in bullous exudative retinal detachment. We evaluated the use of half-dose verteporfin photodynamic therapy for the treatment of bullous retinal detachment in idiopathic central serous chorioretinopathy. CASE PRESENTATION: We report the case of a 51-year-old Chinese man who presented with blurred vision in his right eye and superior visual field defect due to bullous variant of central serous chorioretinopathy. No improvement in vision and retinal detachment was noted after three months of observation and a short course of oral acetazolamide. He was then treated with half-dose verteporfin photodynamic therapy and his visual acuity improved from 20/70 to 20/25 within one month of treatment. Three months after photodynamic therapy, there was complete resolution of sub-retinal fluid and bullous retinal detachment. No recurrence of central serous chorioretinopathy was noted in three years of follow-up. CONCLUSION: We report the beneficial effect of photodynamic therapy with half-dose verteporfin as a treatment option for bullous retinal detachment caused by central serous chorioretinopathy. PMID- 21615894 TI - Curative treatment of oesophageal carcinoma: current options and future developments. AB - Since the 1980s major advances in surgery, radiotherapy and chemotherapy have established multimodal approaches as curative treatment options for oesophageal cancer. In addition the introduction of functional imaging modalities such as PET CT created new opportunities for a more adequate patient selection and therapy response assessment.The majority of oesophageal carcinomas are represented by two histologies: squamous cell carcinoma and adenocarcinoma. In recent years an epidemiological shift towards the latter was observed. From a surgical point of view, adenocarcinomas, which are usually located in the distal third of the oesophagus, may be treated with a transhiatal resection, whereas squamous cell carcinomas, which are typically found in the middle and the upper third, require a transthoracic approach. Since overall survival after surgery alone is poor, multimodality approaches have been developed. At least for patients with locally advanced tumors, surgery alone can no longer be advocated as routine treatment. Nowadays, scientific interest is focused on tumor response to induction radiochemotherapy. A neoadjuvant approach includes the early and accurate assessment of clinical response, optimally performed by repeated PET-CT imaging and endoscopic ultrasound, which may permit early adaption of the therapeutic concept. Patients with SCC that show clinical response by PET CT are considered to have a better prognosis, regardless of whether surgery will be performed or not. In non-responding patients salvage surgery improves survival, especially if complete resection is achieved. PMID- 21615895 TI - Characteristic values of the lumbar load of manual patient handling for the application in workers' compensation procedures. AB - BACKGROUND: The human spine is often exposed to mechanical load in vocational activities especially in combination with lifting, carrying and positioning of heavy objects. This also applies in particular to nursing activities with manual patient handling. In the present study a detailed investigation on the load of the lumbar spine during manual patient handling was performed. METHODS: For a total of 13 presumably endangering activities with transferring a patient, the body movements performed by healthcare workers were recorded and the exerted action forces were determined with regard to magnitude, direction and lateral distribution in the time course with a "measuring bed", a "measuring chair" and a "measuring floor". By the application of biomechanical model calculations the load on the lowest intervertebral disc of the lumbar spine (L5-S1) was determined considering the posture and action force data for every manual patient handling. RESULTS: The results of the investigations reveal the occurrence of high lumbar load during manual patient handling activities, especially in those cases, where awkward postures of the healthcare worker are combined with high action forces caused by the patient's mass. These findings were compared to suitable issues of corresponding investigations provided in the literature. Furthermore measurement based characteristic values of lumbar load were derived for the use in statement procedures concerning the disease no. 2108 of the German list of occupational diseases. CONCLUSIONS: To protect healthcare workers from mechanical overload and the risk of developing a disc-related disease, prevention measures should be compiled. Such measures could include the application of "back-fairer" nursing techniques and the use of "technical" and" small aids" to reduce the lumbar load during manual patient handling. Further studies, concerning these aspects, are necessary. PMID- 21615897 TI - Assessment of replicate bias in 454 pyrosequencing and a multi-purpose read filtering tool. AB - BACKGROUND: Roche 454 pyrosequencing platform is often considered the most versatile of the Next Generation Sequencing technology platforms, permitting the sequencing of large genomes, the analysis of variations or the study of transcriptomes. A recent reported bias leads to the production of multiple reads for a unique DNA fragment in a random manner within a run. This bias has a direct impact on the quality of the measurement of the representation of the fragments using the reads. Other cleaning steps are usually performed on the reads before assembly or alignment. FINDINGS: PyroCleaner is a software module intended to clean 454 pyrosequencing reads in order to ease the assembly process. This program is a free software and is distributed under the terms of the GNU General Public License as published by the Free Software Foundation. It implements several filters using criteria such as read duplication, length, complexity, base pair quality and number of undetermined bases. It also permits to clean flowgram files (.sff) of paired-end sequences generating on one hand validated paired-ends file and the other hand single read file. CONCLUSIONS: Read cleaning has always been an important step in sequence analysis. The pyrocleaner python module is a Swiss knife dedicated to 454 reads cleaning. It includes commonly used filters as well as specialised ones such as duplicated read removal and paired-end read verification. PMID- 21615896 TI - Classically and alternatively activated bone marrow derived macrophages differ in cytoskeletal functions and migration towards specific CNS cell types. AB - BACKGROUND: Macrophages play an important role in neuroinflammatory diseases such as multiple sclerosis (MS) and spinal cord injury (SCI), being involved in both damage and repair. The divergent effects of macrophages might be explained by their different activation status: classically activated (CA/M1), pro inflammatory, macrophages and alternatively activated (AA/M2), growth promoting, macrophages. Little is known about the effect of macrophages with these phenotypes in the central nervous system (CNS) and how they influence pathogenesis. The aim of this study was therefore to determine the characteristics of these phenotypically different macrophages in the context of the CNS in an in vitro setting. RESULTS: Here we show that bone marrow derived CA and AA macrophages have a distinct migratory capacity towards medium conditioned by various cell types of the CNS. AA macrophages were preferentially attracted by the low weight (< 10 kD) fraction of neuronal conditioned medium, while CA macrophages were attracted in higher numbers by astrocyte- and oligodendrocyte conditioned medium. Intrinsic motility was twice as high in AA macrophages compared to CA macrophages. The adhesion to extracellular matrix molecules (ECM) was significantly enhanced in CA macrophages compared to control and AA macrophages. The actin cytoskeleton was differentially organized between CA and AA macrophages, possibly due to greater activity of the GTPases RhoA and Rac in CA macrophages. Phagocytosis of myelin and neuronal fragments was increased in CA macrophages compared to AA macrophages. The increase in myelin phagocytosis was associated with higher expression of CR3/MAC-1 in CA macrophages. CONCLUSION: In conclusion, since AA macrophages are more motile and are attracted by NCM, they are prone to migrate towards neurons in the CNS. CA macrophages have a lower motility and a stronger adhesion to ECM. In neuroinflammatory diseases the restricted migration and motility of CA macrophages might limit lesion size due to bystander damage. PMID- 21615898 TI - Postnatal exposure to PCB 153 and PCB 180, but not to PCB 52, produces changes in activity level and stimulus control in outbred male Wistar Kyoto rats. AB - BACKGROUND: Polychlorinated biphenyls (PCBs) are a class of organic compounds that bioaccumulate due to their chemical stability and lipophilic properties. Humans are prenatally exposed via trans-placental transfer, through breast milk as infants, and through fish, seafood and fatty foods as adolescents and adults. Exposure has several reported effects ranging from developmental abnormalities to cognitive and motor deficiencies. In the present study, three experimental groups of rats were orally exposed to PCBs typically found in human breast milk and then behaviorally tested for changes in measures of stimulus control (percentage lever presses on the reinforcer-producing lever), activity level (responses with IRTs > 0.67 s), and responses with short IRTs (< 0.67 s). METHODS: Male offspring from Wistar Kyoto (WKY/NTac) dams purchased pregnant from Taconic Farms (Germantown, NY) were orally given PCB at around postnatal day 8, 14, and 20 at a dose of 10 mg/kg body weight at each exposure. Three experimental groups were exposed either to PCB 52, PCB 153, or PCB 180. A fourth group fed corn oil only served as controls. From postnatal day 25, for 33 days, the animals were tested for behavioral changes using an operant procedure. RESULTS: PCB exposure did not produce behavioral changes during training when responding was frequently reinforced using a variable interval 3 s schedule. When correct responses were reinforced on a variable interval 180 s schedule, animals exposed to PCB 153 or PCB 180 were less active than controls and animals exposed to PCB 52. Stimulus control was better in animals exposed to PCB 180 than in controls and in the PCB 52 group. Also, the PCB 153 and PCB 180 groups had fewer responses with short IRTs than the PCB 52 group. No effects of exposure to PCB 52 were found when compared to controls. CONCLUSIONS: Exposure to PCBs 153 and 180 produced hypoactivity that continued at least five weeks after the last exposure. No effects of exposure to PCB 52 were observed. PMID- 21615899 TI - Referral of patients with depression to mental health care by Dutch general practitioners: an observational study. AB - BACKGROUND: Depression is a common illness, often treated in primary care. Guidelines provide recommendations for referral to mental health care. Several studies investigated determinants of referral, none investigated guideline criteria as possible determinants.We wanted to evaluate general practitioner's referral of depressed patients to mental health care and to what extent this is in agreement with (Dutch) guideline recommendations. METHODS: We used data of primary care respondents from the Netherlands Study of Depression and Anxiety with major depressive disorder in the past year (n = 478). We excluded respondents with missing data (n = 134). Referral data was collected from electronic patient files between 1 year before and after baseline and self report at baseline and 1-year follow-up. Logistic regression was used to describe association between guideline referral criteria (e.g. perceived need for psychotherapy, suicide risk, severe/chronic depression, antidepressant therapy failure) and referral. RESULTS: A high 58% of depressed patients were referred. Younger patients, those with suicidal tendency, chronic depression or perceived need for psychotherapy were referred more often. Patients who had used >=2 antidepressants or with chronic depression were more often referred to secondary care. Referred respondents met on average more guideline criteria for referral. However, only 8-11% of variance was explained. CONCLUSION: The majority of depressed patients were referred to mental health care. General practitioners take guideline criteria into account in decision making for referral of depressed patients to mental health care. However, other factors play a part, considering the small percentage of variance explained. Further research is necessary to investigate this. PMID- 21615900 TI - Detection of novel chromosome-SCCmec variants in Methicillin Resistant Staphylococcus aureus and their inclusion in PCR based screening. AB - FINDINGS: To facilitate automation, a novel DNA extraction method for MRSA was adopted. The MRSA specific chromosome-SCCmec PCR was adapted, additional primers were added, and the performance was validated. From various laboratories in The Netherlands we received a total of 86 MRSA clinical isolates, that were negative in commercially available tests. We identified 14 MRSA strains with new variant chromosome-SCCmec junctions by sequence analysis. These MRSA strains appeared to carry SCCmec sequences with a high degree of homology to SCC regions of S. epidermidis and S. haemolyticus. All were included for detection in chromosome SCCmec based PCR. BACKGROUND: Efficient management of Methicillin Resistant Staphylococcus aureus (MRSA) in the hospital is needed to prevent dissemination. It is important that MRSA can be rapidly identified, and effective infection control measures can be initiated. Equally important is a rapid MRSA negative report, especially for patients in isolation. For negative screening we implemented fully automated high through-put molecular screening for MRSA. CONCLUSIONS: Fourteen variant chromosome-SCCmec junctions in MRSA, that are not detected in commercially available MRSA detection kits were added to our PCR to detect all currently known variant SCC-mec types of MRSA. PMID- 21615901 TI - The peroxisome proliferator-activated receptor (PPAR) alpha agonist fenofibrate maintains bone mass, while the PPAR gamma agonist pioglitazone exaggerates bone loss, in ovariectomized rats. AB - BACKGROUND: Activation of peroxisome proliferator-activated receptor (PPAR)gamma is associated with bone loss and increased fracture risk, while PPARalpha activation seems to have positive skeletal effects. To further explore these effects we have examined the effect of the PPARalpha agonists fenofibrate and Wyeth 14643, and the PPARgamma agonist pioglitazone, on bone mineral density (BMD), bone architecture and biomechanical strength in ovariectomized rats. METHODS: Fifty-five female Sprague-Dawley rats were assigned to five groups. One group was sham-operated and given vehicle (methylcellulose), the other groups were ovariectomized and given vehicle, fenofibrate, Wyeth 14643 and pioglitazone, respectively, daily for four months. Whole body and femoral BMD were measured by dual X-ray absorptiometry (DXA), and biomechanical testing of femurs, and micro computed tomography (microCT) of the femoral shaft and head, were performed. RESULTS: Whole body and femoral BMD were significantly higher in sham controls and ovariectomized animals given fenofibrate, compared to ovariectomized controls. Ovariectomized rats given Wyeth 14643, maintained whole body BMD at sham levels, while rats on pioglitazone had lower whole body and femoral BMD, impaired bone quality and less mechanical strength compared to sham and ovariectomized controls. In contrast, cortical volume, trabecular bone volume and thickness, and endocortical volume were maintained at sham levels in rats given fenofibrate. CONCLUSIONS: The PPARalpha agonist fenofibrate, and to a lesser extent the PPARaplha agonist Wyeth 14643, maintained BMD and bone architecture at sham levels, while the PPARgamma agonist pioglitazone exaggerated bone loss and negatively affected bone architecture, in ovariectomized rats. PMID- 21615902 TI - Gene expression analysis in lymphoblasts derived from patients with autism spectrum disorder. AB - BACKGROUND: The autism spectrum disorders (ASDs) are complex neurodevelopmental disorders that result in severe and pervasive impairment in the development of reciprocal social interaction and verbal and nonverbal communication skills. In addition, individuals with ASD have stereotypical behavior, interests and activities. Rare mutations of some genes, such as neuroligin (NLGN) 3/4, neurexin (NRXN) 1, SHANK3, MeCP2 and NHE9, have been reported to be associated with ASD. In the present study, we investigated whether alterations in mRNA expression levels of these genes could be found in lymphoblastoid cell lines derived from patients with ASD. METHODS: We measured mRNA expression levels of NLGN3/4, NRXN1, SHANK3, MeCP2, NHE9 and AKT1 in lymphoblastoid cells from 35 patients with ASD and 35 healthy controls, as well as from 45 patients with schizophrenia and 45 healthy controls, using real-time quantitative reverse transcriptase polymerase chain reaction assays. RESULTS: The mRNA expression levels of NLGN3 and SHANK3 normalized by beta-actin or TBP were significantly decreased in the individuals with ASD compared to controls, whereas no difference was found in the mRNA expression level of MeCP2, NHE9 or AKT1. However, normalized NLGN3 and SHANK3 gene expression levels were not altered in patients with schizophrenia, and expression levels of NLGN4 and NRXN1 mRNA were not quantitatively measurable in lymphoblastoid cells. CONCLUSIONS: Our results provide evidence that the NLGN3 and SHANK3 genes may be differentially expressed in lymphoblastoid cell lines from individuals with ASD compared to those from controls. These findings suggest the possibility that decreased mRNA expression levels of these genes might be involved in the pathophysiology of ASD in a substantial population of ASD patients. PMID- 21615903 TI - Sampling strategies for accurate computational inferences of gametic phase across highly polymorphic major histocompatibility complex loci. AB - BACKGROUND: Genes of the Major Histocompatibility Complex (MHC) are very popular genetic markers among evolutionary biologists because of their potential role in pathogen confrontation and sexual selection. However, MHC genotyping still remains challenging and time-consuming in spite of substantial methodological advances. Although computational haplotype inference has brought into focus interesting alternatives, high heterozygosity, extensive genetic variation and population admixture are known to cause inaccuracies. We have investigated the role of sample size, genetic polymorphism and genetic structuring on the performance of the popular Bayesian PHASE algorithm. To cover this aim, we took advantage of a large database of known genotypes (using traditional laboratory based techniques) at single MHC class I (N = 56 individuals and 50 alleles) and MHC class II B (N = 103 individuals and 62 alleles) loci in the lesser kestrel Falco naumanni. FINDINGS: Analyses carried out over real MHC genotypes showed that the accuracy of gametic phase reconstruction improved with sample size as a result of the reduction in the allele to individual ratio. We then simulated different data sets introducing variations in this parameter to define an optimal ratio. CONCLUSIONS: Our results demonstrate a critical influence of the allele to individual ratio on PHASE performance. We found that a minimum allele to individual ratio (1:2) yielded 100% accuracy for both MHC loci. Sampling effort is therefore a crucial step to obtain reliable MHC haplotype reconstructions and must be accomplished accordingly to the degree of MHC polymorphism. We expect our findings provide a foothold into the design of straightforward and cost-effective genotyping strategies of those MHC loci from which locus-specific primers are available. PMID- 21615904 TI - Epidemiology of acute and chronic hepatitis B virus infection in Norway, 1992 2009. AB - BACKGROUND: Norway is classified as a low prevalence country for hepatitis B virus infection. Vaccination is only recommended for risk groups (intravenous drug users (IDUs), Men who have Sex with Men (MSM), immigrants and contacts of known carriers). We describe the epidemiology of reported cases of hepatitis B in Norway, during the years 1992-2009 in order to assess the validity of current risk groups and recommend preventive measures. METHODS: We used case based data from the national surveillance system on acute and chronic hepatitis B. The Norwegian Statistics Bureau provided population and migration data and the Norwegian Institute for Alcohol and Drug Research the estimated number of active IDUs between 2002-2007. Incidence rates (IR) and incidence rate ratios (IRR) for acute hepatitis B and notification rates (NR) and notification rate ratios (NRR) for chronic hepatitis B with 95% confidence intervals were calculated. RESULTS: The annual IR of acute hepatitis B ranged from 0.7/100,000 (1992) to 10.6/100,000 (1999). Transmission occurred mainly among IDUs (64%) or through sexual contact (24%). The risk of acquiring acute hepatitis B was highest in people aged 20-29 (IRR = 6.6 [3.3-13.3]), and in males (IRR = 2.4 [1.7-3.3]). We observed two peaks of newly reported chronic hepatitis B cases in 2003 and 2009 (NR = 17.6/100,000 and 17.4/100,000, respectively). Chronic hepatitis B was more likely to be diagnosed among immigrants than among Norwegians (NRR = 93 [71.9-120.6]), and among those 20-29 compared to those 50-59 (NRR = 5.2 [3.5-7.9]). CONCLUSIONS: IDUs remain the largest risk group for acute hepatitis B. The observed peaks of chronic hepatitis B are related to increased immigration from high endemic countries and screening and vaccination of these groups is important to prevent further spread of infection. Universal screening of pregnant women should be introduced. A universal vaccination strategy should be considered, given the high cost of reaching the target populations. We recommend evaluating the surveillance system for hepatitis B as well as the effectiveness of screening and vaccinating immigrant populations. PMID- 21615905 TI - Introduction of a new model for time-continuous and non-contact investigations of in-vitro thrombolysis under physiological flow conditions. AB - BACKGROUND: Thrombolysis is a dynamic and time-dependent process influenced by the haemodynamic conditions. Currently there is no model that allows for time continuous, non-contact measurements under physiological flow conditions. The aim of this work was to introduce such a model. METHODS: The model is based on a computer-controlled pump providing variable constant or pulsatile flows in a tube system filled with blood substitute. Clots can be fixed in a custom-built clot carrier within the tube system. The pressure decline at the clot carrier is measured as a novel way to measure lysis of the clot. With different experiments the hydrodynamic properties and reliability of the model were analyzed. Finally, the lysis rate of clots generated from human platelet rich plasma (PRP) was measured during a one hour combined application of diagnostic ultrasound (2 MHz, 0.179 W/cm2) and a thrombolytic agent (rt-PA) as it is commonly used for clinical sonothrombolysis treatments. RESULTS: All hydrodynamic parameters can be adjusted and measured with high accuracy. First experiments with sonothrombolysis demonstrated the feasibility of the model despite low lysis rates. CONCLUSIONS: The model allows to adjust accurately all hydrodynamic parameters affecting thrombolysis under physiological flow conditions and for non-contact, time continuous measurements. Low lysis rates of first sonothrombolysis experiments are primarily attributable to the high stability of the used PRP-clots. PMID- 21615907 TI - Serum Mullerian inhibiting substance levels are lower in premenopausal women with breast precancer and cancer. AB - BACKGROUND: In preclinical studies, mullerian inhibiting substance (MIS) has a protective affect against breast cancer. Our objective was to determine whether serum MIS concentrations were associated with cancerous or precancerous lesions. Blood from 30 premenopausal women was collected and serum extracted prior to their undergoing breast biopsy to assess a suspicious lesion found on imaging or physical examination. Based on biopsy results, the serum specimens were grouped as cancer (invasive or ductal carcinoma in situ), precancer (atypical hyperplasia or lobular carcinoma in situ), or benign. FINDINGS: Serum from women with cancer and precancer (p = .0009) had lower MIS levels than serum from women with benign disease. CONCLUSION: Our findings provide preliminary evidence for MIS being associated with current breast cancer risk, which should be validated in a larger population. PMID- 21615908 TI - Characterization of the expression, promoter activity and molecular architecture of fibin. AB - BACKGROUND: Fibin was initially discovered as a secreted signal molecule essential for pectoral fin bud initiation in zebrafish. Currently, there is little information about the molecular architecture and biological relevance of fibin in humans and other mammals. RESULTS: Fibin is expressed in cerebellum, skeletal muscle and many other embryonic and adult mouse tissues suggesting not only a role during embryonic development but also in adult functions. A 2.5-kbp genomic sequence fragment upstream of the coding sequence is sufficient to drive and regulate fibin expression through stimulation by glucocorticoids, activators of the protein kinase C signalling pathways and manganese ions. Fibin is an evolutionarily conserved protein, carries a cleavable signal peptide (amino acids 1-18) and is glycosylated at Asn30. The two conserved cysteines participate in intermolecular disulfide bond and multimer formation. Although fibin displays all features of a secretory protein, it is mostly retained in the endoplasmic reticulum when heterologously expressed. CONCLUSION: Fibin is functionally relevant during embryogenesis and adult life. Its expression is regulated by a number of cellular signalling pathways and the protein is routed via the secretory pathway. However, proper secretion presumably requires an unknown covalently-linked or associated co-factor. PMID- 21615906 TI - Modelling the global constraints of temperature on transmission of Plasmodium falciparum and P. vivax. AB - BACKGROUND: Temperature is a key determinant of environmental suitability for transmission of human malaria, modulating endemicity in some regions and preventing transmission in others. The spatial modelling of malaria endemicity has become increasingly sophisticated and is now central to the global scale planning, implementation, and monitoring of disease control and regional efforts towards elimination, but existing efforts to model the constraints of temperature on the malaria landscape at these scales have been simplistic. Here, we define an analytical framework to model these constraints appropriately at fine spatial and temporal resolutions, providing a detailed dynamic description that can enhance large scale malaria cartography as a decision-support tool in public health. RESULTS: We defined a dynamic biological model that incorporated the principal mechanisms of temperature dependency in the malaria transmission cycle and used it with fine spatial and temporal resolution temperature data to evaluate time series of temperature suitability for transmission of Plasmodium falciparum and P. vivax throughout an average year, quantified using an index proportional to the basic reproductive number. Time-series were calculated for all 1 km resolution land pixels globally and were summarised to create high-resolution maps for each species delineating those regions where temperature precludes transmission throughout the year. Within suitable zones we mapped for each pixel the number of days in which transmission is possible and an integrated measure of the intensity of suitability across the year. The detailed evaluation of temporal suitability dynamics provided by the model is visualised in a series of accompanying animations. CONCLUSIONS: These modelled products, made available freely in the public domain, can support the refined delineation of populations at risk; enhance endemicity mapping by offering a detailed, dynamic, and biologically driven alternative to the ubiquitous empirical incorporation of raw temperature data in geospatial models; and provide a rich spatial and temporal platform for future biological modelling studies. PMID- 21615909 TI - Quality of tumor lysates used for pulsing dendritic cells is influenced by the method used to harvest adherent tumor cells. AB - BACKGROUND: Lysates from tumor cells are reported to induce maturation of dendritic cells (DCs) and are used in clinical settings for DC-based vaccination against solid tumors. Nevertheless, the maturation inducing effect of tumor lysates on DCs is discussed controversially and the efficacy of tumor vaccines varies significantly. FINDINGS: Using three individual adherent colorectal tumor cell lines we also faced the difficulty to obtain consistent results regarding maturation inducing effect of tumor lysates on DCs. Therefore, we compared different methods to prepare tumor cell lysate and could demonstrate that trypsinizing as a method to harvest adherent tumor cells has a significant negative impact on biologic activity of tumor lysates. Specifically, we assessed induction of maturation markers CD40, CD80, and CD86 on DCs which were treated with differently prepared lysates. CONCLUSIONS: Trypsinizing is a very common way of harvesting adherent cells from culture flasks. Our results shall call investigators' attention to the enzymatic activity of trypsin degrading some possibly important proteins on the surface of cultured cells. Specifically for DC based vaccination against tumor antigens investigators should avoid trypsin. PMID- 21615912 TI - qtl.outbred: Interfacing outbred line cross data with the R/qtl mapping software. AB - BACKGROUND: qtl.outbred is an extendible interface in the statistical environment, R, for combining quantitative trait loci (QTL) mapping tools. It is built as an umbrella package that enables outbred genotype probabilities to be calculated and/or imported into the software package R/qtl. FINDINGS: Using qtl.outbred, the genotype probabilities from outbred line cross data can be calculated by interfacing with a new and efficient algorithm developed for analyzing arbitrarily large datasets (included in the package) or imported from other sources such as the web-based tool, GridQTL. CONCLUSION: qtl.outbred will improve the speed for calculating probabilities and the ability to analyse large future datasets. This package enables the user to analyse outbred line cross data accurately, but with similar effort than inbred line cross data. PMID- 21615911 TI - Acute hospital admissions among nursing home residents: a population-based observational study. AB - BACKGROUND: Nursing home residents are prone to acute illness due to their high age, underlying illnesses and immobility. We examined the incidence of acute hospital admissions among nursing home residents versus the age-matched community dwelling population in a geographically defined area during a two years period. The hospital stays of the nursing home population are described according to diagnosis, length of stay and mortality. Similar studies have previously not been reported in Scandinavia. METHODS: The acute hospitalisations of the nursing home residents were identified through ambulance records. These were linked to hospital patient records for inclusion of demographics, diagnosis at discharge, length of stay and mortality. Incidence of hospitalisation was calculated based on patient-time at risk. RESULTS: The annual hospital admission incidence was 0.62 admissions per person-year among the nursing home residents and 0.26 among the community dwellers. In the nursing home population we found that dominant diagnoses were respiratory diseases, falls-related and circulatory diseases, accounting for 55% of the cases. The median length of stay was 3 days (interquartile range = 4). The in-hospital mortality rate was 16% and 30 day mortality after discharge 30%. CONCLUSION: Acute hospital admission rate among nursing home residents was high in this Scandinavian setting. The pattern of diagnoses causing the admissions appears to be consistent with previous research. The in-hospital and 30 day mortality rates are high. PMID- 21615910 TI - Interplay of recombination and selection in the genomes of Chlamydia trachomatis. AB - BACKGROUND: Chlamydia trachomatis is an obligate intracellular bacterial parasite, which causes several severe and debilitating diseases in humans. This study uses comparative genomic analyses of 12 complete published C. trachomatis genomes to assess the contribution of recombination and selection in this pathogen and to understand the major evolutionary forces acting on the genome of this bacterium. RESULTS: The conserved core genes of C. trachomatis are a large proportion of the pan-genome: we identified 836 core genes in C. trachomatis out of a range of 874-927 total genes in each genome. The ratio of recombination events compared to mutation (rho/theta) was 0.07 based on ancestral reconstructions using the ClonalFrame tool, but recombination had a significant effect on genetic diversification (r/m=0.71). The distance-dependent decay of linkage disequilibrium also indicated that C. trachomatis populations behaved intermediately between sexual and clonal extremes. Fifty-five genes were identified as having a history of recombination and 92 were under positive selection based on statistical tests. Twenty-three genes showed evidence of being under both positive selection and recombination, which included genes with a known role in virulence and pathogencity (e.g., ompA, pmps, tarp). Analysis of inter-clade recombination flux indicated non-uniform currents of recombination between clades, which suggests the possibility of spatial population structure in C. trachomatis infections. CONCLUSIONS: C. trachomatis is the archetype of a bacterial species where recombination is relatively frequent yet gene gains by horizontal gene transfer (HGT) and losses (by deletion) are rare. Gene conversion occurs at sites across the whole C. trachomatis genome but may be more often fixed in genes that are under diversifying selection. Furthermore, genome sequencing will reveal patterns of serotype specific gene exchange and selection that will generate important research questions for understanding C. trachomatis pathogenesis. REVIEWERS: This article was reviewed by Dr. Jeremy Selengut, Dr. Lee S. Katz (nominated by Dr. I. King Jordan) and Dr. Arcady Mushegian. PMID- 21615913 TI - A novel and well-defined benchmarking method for second generation read mapping. AB - BACKGROUND: Second generation sequencing technologies yield DNA sequence data at ultra high-throughput. Common to most biological applications is a mapping of the reads to an almost identical or highly similar reference genome. The assessment of the quality of read mapping results is not straightforward and has not been formalized so far. Hence, it has not been easy to compare different read mapping approaches in a unified way and to determine which program is the best for what task. RESULTS: We present a new benchmark method, called Rabema (Read Alignment BEnchMArk), for read mappers. It consists of a strict definition of the read mapping problem and of tools to evaluate the result of arbitrary read mappers supporting the SAM output format. CONCLUSIONS: We show the usefulness of the benchmark program by performing a comparison of popular read mappers. The tools supporting the benchmark are licensed under the GPL and available from http://www.seqan.de/projects/rabema.html. PMID- 21615914 TI - Prevalence of self-reported finger deformations and occupational risk factors among professional cooks: a cross-sectional study. AB - BACKGROUND: Previous studies have pointed out that the school lunch workers in Japan are suffering from work-related disorders including finger deformations. The purpose of this study was to investigate the prevalence of self-reported finger deformations and the association with job-related risk factors. METHODS: A cross-sectional questionnaire study of 5,719 subjects (response rate: 81%, 982 men and 4,737 women) was undertaken during September 2003 to February 2004. RESULTS: Finger deformations were found among 11.7% of the men and 35.6% of the women studied, with significant differences among sex, age and sex-age groups. For both men and women the pattern of finger deformations across the hand was similar for the right and the left hand. For women, the deformations were found in about 10% of the distal interphalangeal joints of all fingers. Based on multiple logistic regression analyses, the factors female sex, age, the number of cooked lunches per cook and cooking activities were independently associated with the prevalence of finger deformations. High prevalence odds ratios were found for those frequently carrying or using tools by hands such as delivering containers, distributing meals, preparing dishes, washing equipment, cutting and stirring foods. CONCLUSIONS: Among the school lunch workers studied, women had a higher prevalence of finger deformations on all joints of both hands. Various cooking tasks were associated with the prevalence of finger deformations. The results suggest that improvements in working conditions are important for preventing work related disorders such as finger deformations. PMID- 21615915 TI - Common somatic symptoms, causal attributions of somatic symptoms and psychiatric morbidity in a cross-sectional community study in Santiago, Chile. AB - BACKGROUND: It has been suggested that individuals from non-western countries tend to deny or mask psychological symptoms of common mental disorders and to present with somatic symptoms. The aim of the present paper was to investigate the association between common mental disorders and somatic symptoms in a representative sample of the general population of Santiago, Chile FINDINGS: This was a cross-sectional study of a stratified random sample of 3807 subjects living in private households in Santiago, Chile. Psychiatric disorders were assessed with the revised Clinical Interview Schedule. We found a strong association between the presence of somatic symptoms and psychiatric disorders (odds ratio 3.20, [95% confidence interval 2.52 - 4.05]). In addition, subjects who attributed their somatic symptoms to psychological or mixed psychological/physical causes were more likely to be cases compared to subjects who made physical attributions only (odds ratios 7.10 [95% CI 4.49-11.25] and 9.27 [6.00-14.34] respectively. CONCLUSIONS: The study confirms previous observations from more selected samples that subjects of Hispanic origin are generally aware of the link between somatic symptoms and psychological ill-health and do not hide or "mask" their psychological symptoms. PMID- 21615916 TI - Tryptophan degradation in women with breast cancer: a pilot study. AB - BACKGROUND: Altered tryptophan metabolism and indoleamine 2,3-dioxygenase activity are linked to cancer development and progression. In addition, these biological factors have been associated with the development and severity of neuropsychiatric syndromes, including major depressive disorder. However, this biological mechanism associated with both poor disease outcomes and adverse neuropsychiatric symptoms has received little attention in women with breast cancer. Therefore, a pilot study was undertaken to compare levels of tryptophan and other proteins involved in tryptophan degradation in women with breast cancer to women without cancer, and secondarily, to examine levels in women with breast caner over the course of chemotherapy. FINDINGS: Blood samples were collected from women with a recent diagnosis of breast cancer (n = 33) before their first cycle of chemotherapy and after their last cycle of chemotherapy. The comparison group (n = 24) provided a blood sample prior to breast biopsy. Plasma concentrations of tryptophan, kynurenine, and tyrosine were determined. The kynurenine to tryptophan ratio (KYN/TRP) was used to estimate indoleamine 2,3 dioxygenase activity. On average, the women with breast cancer had lower levels of tryptophan, elevated levels of kynurenine and tyrosine and an increased KYN/TRP ratio compared to women without breast cancer. There was a statistically significant difference between the two groups in the KYN/TRP ratio (p = 0.036), which remained elevated in women with breast cancer throughout the treatment trajectory. CONCLUSIONS: The findings of this pilot study suggest that increased tryptophan degradation may occur in women with early-stage breast cancer. Given the multifactorial consequences of increased tryptophan degradation in cancer outcomes and neuropsychiatric symptom manifestation, this biological mechanism deserves broader attention in women with breast cancer. PMID- 21615917 TI - Noninvasive monitoring of cardiac function in a chronic ischemic heart failure model in the rat: assessment with tissue Doppler and non-Doppler 2D strain echocardiography. AB - OBJECTIVES: Feasibility of noninvasive monitoring of cardiac function after surgically induced ischemic cardiomyopathy with tissue Doppler and non-Doppler 2D strain echocardiography in rats. BACKGROUND: The optimal method for quantitative assessment of global and regional ventricular function in rats with chronic heart failure for research purposes remains unclear. METHODS: 20 rats underwent suture ligation of the left anterior descending coronary artery via a left thoracotomy to induce ischemic cardiomyopathy. Echocardiographic examination with estimation of left ventricular wall thickness, diameters, fractional shortening, ejection fraction, wall velocities as well as radial strain were performed before and 4 weeks after surgery. RESULTS: Mean LVEF decreased from 70 +/- 6% to 40 +/- 8% (p < 0.0001) one month after the operation. LVEDD increased from 7 +/- 1 mm to 9 +/- 1 mm (p < 0.0001), systolic anterior velocity decreased from 0.79 +/- 0.25 cm/s to 0.18 +/- 0.19 cm/s (p < 0.0001). Radial 2D strain was significantly reduced after myocardial infarction of the septal (18.2 +/- 6.6% vs 7.0 +/- 5.9%, p < 0.001), anteroseptal (17.3 +/- 5.2% vs 4.6 +/- 3.0%, p < 0.0001), anterior (18.9 +/- 5.9% vs 5.6 +/- 2.5%, p < 0.0001), lateral (21.4 +/- 4.9% vs 8.1 +/- 3.5%, p < 0.0001) as well as posterior myocardial segments (19.3 +/- 5.2% vs 15.4 +/- 5.5%, p < 0.01). Inferior segments (19.2 +/- 7.9% vs 17.8 +/- 7.9%, ns) did not change at all. CONCLUSION: It is feasible to assess dimensions, global function, and regional contractility with echocardiography in rats suffering from chronic heart failure after myocardial infarction. Particularly regional function can be exactly evaluated if tissue Doppler and 2D strain is used. PMID- 21615918 TI - Methylation levels of the "long interspersed nucleotide element-1" repetitive sequences predict survival of melanoma patients. AB - BACKGROUND: The prognosis of cutaneous melanoma (CM) differs for patients with identical clinico-pathological stage, and no molecular markers discriminating the prognosis of stage III individuals have been established. Genome-wide alterations in DNA methylation are a common event in cancer. This study aimed to define the prognostic value of genomic DNA methylation levels in stage III CM patients. METHODS: Overall level of genomic DNA methylation was measured using bisulfite pyrosequencing at three CpG sites (CpG1, CpG2, CpG3) of the Long Interspersed Nucleotide Element-1 (LINE-1) sequences in short-term CM cultures from 42 stage IIIC patients. The impact of LINE-1 methylation on overall survival (OS) was assessed using Cox regression and Kaplan-Meier analysis. RESULTS: Hypomethylation (i.e., methylation below median) at CpG2 and CpG3 sites significantly associated with improved prognosis of CM, CpG3 showing the strongest association. Patients with hypomethylated CpG3 had increased OS (P = 0.01, log-rank = 6.39) by Kaplan Meyer analysis. Median OS of patients with hypomethylated or hypermethylated CpG3 were 31.9 and 11.5 months, respectively. The 5 year OS for patients with hypomethylated CpG3 was 48% compared to 7% for patients with hypermethylated sequences. Among the variables examined by Cox regression analysis, LINE-1 methylation at CpG2 and CpG3 was the only predictor of OS (Hazard Ratio = 2.63, for hypermethylated CpG3; 95% Confidence Interval: 1.21-5.69; P = 0.01). CONCLUSION: LINE-1 methylation is identified as a molecular marker of prognosis for CM patients in stage IIIC. Evaluation of LINE-1 promises to represent a key tool for driving the most appropriate clinical management of stage III CM patients. PMID- 21615919 TI - Molecular cytogenetic characterization of canine histiocytic sarcoma: A spontaneous model for human histiocytic cancer identifies deletion of tumor suppressor genes and highlights influence of genetic background on tumor behavior. AB - BACKGROUND: Histiocytic malignancies in both humans and dogs are rare and poorly understood. While canine histiocytic sarcoma (HS) is uncommon in the general domestic dog population, there is a strikingly high incidence in a subset of breeds, suggesting heritable predisposition. Molecular cytogenetic profiling of canine HS in these breeds would serve to reveal recurrent DNA copy number aberrations (CNAs) that are breed and/or tumor associated, as well as defining those shared with human HS. This process would identify evolutionarily conserved cytogenetic changes to highlight regions of particular importance to HS biology. METHODS: Using genome wide array comparative genomic hybridization we assessed CNAs in 104 spontaneously occurring HS from two breeds of dog exhibiting a particularly elevated incidence of this tumor, the Bernese Mountain Dog and Flat Coated Retriever. Recurrent CNAs were evaluated further by multicolor fluorescence in situ hybridization and loss of heterozygosity analyses. Statistical analyses were performed to identify CNAs associated with tumor location and breed. RESULTS: Almost all recurrent CNAs identified in this study were shared between the two breeds, suggesting that they are associated more with the cancer phenotype than with breed. A subset of recurrent genomic imbalances suggested involvement of known cancer associated genes in HS pathogenesis, including deletions of the tumor suppressor genes CDKN2A/B, RB1 and PTEN. A small number of aberrations were unique to each breed, implying that they may contribute to the major differences in tumor location evident in these two breeds. The most highly recurrent canine CNAs revealed in this study are evolutionarily conserved with those reported in human histiocytic proliferations, suggesting that human and dog HS share a conserved pathogenesis. CONCLUSIONS: The breed associated clinical features and DNA copy number aberrations exhibited by canine HS offer a valuable model for the human counterpart, providing additional evidence towards elucidation of the pathophysiological and genetic mechanisms associated with histiocytic malignancies. Extrapolation of data derived from canine histiocytic disorders to human histiocytic proliferation may help to further our understanding of the propagation and cancerization of histiocytic cells, contributing to development of new and effective therapeutic modalities for both species. PMID- 21615920 TI - Transcriptional regulatory program in wild-type and retinoblastoma gene-deficient mouse embryonic fibroblasts during adipocyte differentiation. AB - BACKGROUND: Although many molecular regulators of adipogenesis have been identified a comprehensive catalogue of components is still missing. Recent studies showed that the retinoblastoma protein (pRb) was expressed in the cell cycle and late cellular differentiation phase during adipogenesis. To investigate this dual role of pRb in the early and late stages of adipogenesis we used microarrays to perform a comprehensive systems-level analysis of the common transcriptional program of the classic 3T3-L1 preadipocyte cell line, wild-type mouse embryonic fibroblasts (MEFs), and retinoblastoma gene-deficient MEFs (Rb-/- MEFs). FINDINGS: Comparative analysis of the expression profiles of 3T3-L1 cells and wild-type MEFs revealed genes involved specifically in early regulation of the adipocyte differentiation as well as secreted factors and signaling molecules regulating the later phase of differentiation. In an attempt to identify transcription factors regulating adipogenesis, bioinformatics analysis of the promoters of coordinately and highly expressed genes was performed. We were able to identify a number of high-confidence target genes for follow-up experimental studies. Additionally, combination of experimental data and computational analyses pinpointed a feedback-loop between Pparg and Foxo1.To analyze the effects of the retinoblastoma protein at the transcriptional level we chose a perturbated system (Rb-/- MEFs) for comparison to the transcriptional program of wild-type MEFs. Gene ontology analysis of 64 deregulated genes showed that the Rb /- MEF model exhibits a brown(-like) adipocyte phenotype. Additionally, the analysis results indicate a different or additional role for pRb family member involvement in the lineage commitment. CONCLUSION: In this study a number of commonly modulated genes during adipogenesis in 3T3-L1 cells and MEFs, potential transcriptional regulation mechanisms, and differentially regulated targets during adipocyte differentiation of Rb-/- MEFs could be identified. These data and the analysis provide a starting point for further experimental studies to identify target genes for pharmacological intervention and ultimately remodeling of white adipose tissue into brown adipose tissue. PMID- 21615921 TI - Transgenic overexpression of miR-133a in skeletal muscle. AB - BACKGROUND: MicroRNAs (miRNAs) are a class of non-coding regulatory RNAs of ~22 nucleotides in length. miRNAs regulate gene expression post-transcriptionally, primarily by associating with the 3' untranslated region (UTR) of their regulatory target mRNAs. Recent work has begun to reveal roles for miRNAs in a wide range of biological processes, including cell proliferation, differentiation and apoptosis. Many miRNAs are expressed in cardiac and skeletal muscle, and dysregulated miRNA expression has been correlated with muscle-related disorders. We have previously reported that the expression of muscle-specific miR-1 and miR 133 is induced during skeletal muscle differentiation and miR-1 and miR-133 play central regulatory roles in myoblast proliferation and differentiation in vitro. METHODS: In this study, we measured the expression of miRNAs in the skeletal muscle of mdx mice, an animal model for human muscular dystrophy. We also generated transgenic mice to overexpress miR-133a in skeletal muscle. RESULTS: We examined the expression of miRNAs in the skeletal muscle of mdx mice. We found that the expression of muscle miRNAs, including miR-1a, miR-133a and miR-206, was up-regulated in the skeletal muscle of mdx mice. In order to further investigate the function of miR-133a in skeletal muscle in vivo, we have created several independent transgenic founder lines. Surprisingly, skeletal muscle development and function appear to be unaffected in miR-133a transgenic mice. CONCLUSIONS: Our results indicate that miR-133a is dispensable for the normal development and function of skeletal muscle. PMID- 21615923 TI - GENIE: a software package for gene-gene interaction analysis in genetic association studies using multiple GPU or CPU cores. AB - BACKGROUND: Gene-gene interaction in genetic association studies is computationally intensive when a large number of SNPs are involved. Most of the latest Central Processing Units (CPUs) have multiple cores, whereas Graphics Processing Units (GPUs) also have hundreds of cores and have been recently used to implement faster scientific software. However, currently there are no genetic analysis software packages that allow users to fully utilize the computing power of these multi-core devices for genetic interaction analysis for binary traits. FINDINGS: Here we present a novel software package GENIE, which utilizes the power of multiple GPU or CPU processor cores to parallelize the interaction analysis. GENIE reads an entire genetic association study dataset into memory and partitions the dataset into fragments with non-overlapping sets of SNPs. For each fragment, GENIE analyzes: 1) the interaction of SNPs within it in parallel, and 2) the interaction between the SNPs of the current fragment and other fragments in parallel. We tested GENIE on a large-scale candidate gene study on high density lipoprotein cholesterol. Using an NVIDIA Tesla C1060 graphics card, the GPU mode of GENIE achieves a speedup of 27 times over its single-core CPU mode run. CONCLUSIONS: GENIE is open-source, economical, user-friendly, and scalable. Since the computing power and memory capacity of graphics cards are increasing rapidly while their cost is going down, we anticipate that GENIE will achieve greater speedups with faster GPU cards. Documentation, source code, and precompiled binaries can be downloaded from http://www.cceb.upenn.edu/~mli/software/GENIE/. PMID- 21615922 TI - Cardiac lipid content is unresponsive to a physical activity training intervention in type 2 diabetic patients, despite improved ejection fraction. AB - BACKGROUND: Increased cardiac lipid content has been associated with diabetic cardiomyopathy. We recently showed that cardiac lipid content is reduced after 12 weeks of physical activity training in healthy overweight subjects. The beneficial effect of exercise training on cardiovascular risk is well established and the decrease in cardiac lipid content with exercise training in healthy overweight subjects was accompanied by improved ejection fraction. It is yet unclear whether diabetic patients respond similarly to physical activity training and whether a lowered lipid content in the heart is necessary for improvements in cardiac function. Here, we investigated whether exercise training is able to lower cardiac lipid content and improve cardiac function in type 2 diabetic patients. METHODS: Eleven overweight-to-obese male patients with type 2 diabetes mellitus (age: 58.4 +/- 0.9 years, BMI: 29.9 +/- 0.01 kg/m2) followed a 12-week training program (combination endurance/strength training, three sessions/week). Before and after training, maximal whole body oxygen uptake (VO2max) and insulin sensitivity (by hyperinsulinemic, euglycemic clamp) was determined. Systolic function was determined under resting conditions by CINE-MRI and cardiac lipid content in the septum of the heart by Proton Magnetic Resonance Spectroscopy. RESULTS: VO2max increased (from 27.1 +/- 1.5 to 30.1 +/- 1.6 ml/min/kg, p = 0.001) and insulin sensitivity improved upon training (insulin stimulated glucose disposal (delta Rd of glucose) improved from 5.8 +/- 1.9 to 10.3 +/- 2.0 MUmol/kg/min, p = 0.02. Left-ventricular ejection fraction improved after training (from 50.5 +/- 2.0 to 55.6 +/- 1.5%, p = 0.01) as well as cardiac index and cardiac output. Unexpectedly, cardiac lipid content in the septum remained unchanged (from 0.80 +/- 0.22% to 0.95 +/- 0.21%, p = 0.15). CONCLUSIONS: Twelve weeks of progressive endurance/strength training was effective in improving VO2max, insulin sensitivity and cardiac function in patients with type 2 diabetes mellitus. However, cardiac lipid content remained unchanged. These data suggest that a decrease in cardiac lipid content in type 2 diabetic patients is not a prerequisite for improvements in cardiac function. TRIAL REGISTRATION: ISRCTN: ISRCTN43780395. PMID- 21615924 TI - High quality and quantity Genome-wide germline genotypes from FFPE normal tissue. AB - BACKGROUND: Although collections of formalin fixed paraffin embedded (FFPE) samples exist, sometimes representing decades of stored samples, they have not typically been utilized to their full potential. Normal tissue from such samples would be extremely valuable for generation of genotype data for individuals who cannot otherwise provide a DNA sample. FINDINGS: We extracted DNA from normal tissue identified in FFPE tissue blocks from prostate surgery and obtained complete genome wide genotype data for over 500,000 SNP markers for these samples, and for DNA extracted from whole blood for 2 of the cases, for comparison.Four of the five FFPE samples of varying age and amount of tissue had identifiable normal tissue. We obtained good quality genotype data for between 89 and 99% of all SNP markers for the 4 samples from FFPE. Concordance rates of over 99% were observed for the 2 samples with DNA from both FFPE and from whole blood. CONCLUSIONS: DNA extracted from normal FFPE tissue provides excellent quality and quantity genome-wide genotyping data representing germline DNA, sufficient for both linkage and association analyses. This allows genetic analysis of informative individuals who are no longer available for sampling in genetic studies. PMID- 21615925 TI - The dynamics of E1A in regulating networks and canonical pathways in quiescent cells. AB - BACKGROUND: Adenoviruses force quiescent cells to re-enter the cell cycle to replicate their DNA, and for the most part, this is accomplished after they express the E1A protein immediately after infection. In this context, E1A is believed to inactivate cellular proteins (e.g., p130) that are known to be involved in the silencing of E2F-dependent genes that are required for cell cycle entry. However, the potential perturbation of these types of genes by E1A relative to their functions in regulatory networks and canonical pathways remains poorly understood. FINDINGS: We have used DNA microarrays analyzed with Bayesian ANOVA for microarray (BAM) to assess changes in gene expression after E1A alone was introduced into quiescent cells from a regulated promoter. Approximately 2,401 genes were significantly modulated by E1A, and of these, 385 and 1033 met the criteria for generating networks and functional and canonical pathway analysis respectively, as determined by using Ingenuity Pathway Analysis software. After focusing on the highest-ranking cellular processes and regulatory networks that were responsive to E1A in quiescent cells, we observed that many of the up-regulated genes were associated with DNA replication, the cell cycle and cellular compromise. We also identified a cadre of up regulated genes with no previous connection to E1A; including genes that encode components of global DNA repair systems and DNA damage checkpoints. Among the down-regulated genes, we found that many were involved in cell signalling, cell movement, and cellular proliferation. Remarkably, a subset of these was also associated with p53 independent apoptosis, and the putative suppression of this pathway may be necessary in the viral life cycle until sufficient progeny have been produced. CONCLUSIONS: These studies have identified for the first time a large number of genes that are relevant to E1A's activities in promoting quiescent cells to re enter the cell cycle in order to create an optimum environment for adenoviral replication. PMID- 21615927 TI - A post-labeling method for multiplexed and multicolored genotyping analysis of SSR, indel and SNP markers in single tube with bar-coded split tag (BStag). AB - BACKGROUND: Genotyping analysis using capillary DNA sequencing with fluorescently labeled primer pairs obtained by polymerase chain reaction (PCR) is widely used, but is expensive. The post-PCR labeling method using fluorescently labeled short oligonucleotides and nested PCR of the amplified product obtained from unlabeled primer pairs is a simple and inexpensive alternative. However, previously reported protocols often produced spurious peaks or inconsistent amplification under multiplexed analysis as a result of simultaneous progress of both the amplification and labeling reactions and local homology of the attached tag sequence. RESULTS: A set of 16 bp-long oligonucleotide sequences termed bar-coded split tag (BStag), comprising a common basal region, a three-nucleotide 'bar code' sequence, and a mismatched nucleotide at the middle position were designed for selective post-PCR labeling. The BStag was attached at the 5' end of the forward primer of interest. The melting temperature of the BStag was low enough to separate the labeling reaction from initial PCR amplification, and each sequence was minimally divergent but maintained maximum selectivity. Post-PCR labeling of the amplified product was achieved by extending for three cycles at a lower annealing temperature after the conventional amplification program with the appropriate fluorescently labeled BStag primer. No amplification was confirmed with BStag primers for 12 plant species. The electropherogram of the labeled product obtained using this method was consistent with that of prelabeled primer, except for their apparent size. CONCLUSIONS: BStag enabled multiplexed post-PCR labeling of simple sequence repeat or insertion/deletion markers with different dyes in a single tube. BStag in conjunction with locus specific oligo and allele specific oligo was also useful for single nucleotide polymorphism analysis. The labeling protocol was simple and no additional operation was required. Single tube multiplexed post-PCR labeling is useful for a wide variety of genotyping studies with maximal flexibility and minimal costs. PMID- 21615926 TI - Analysis of common bean expressed sequence tags identifies sulfur metabolic pathways active in seed and sulfur-rich proteins highly expressed in the absence of phaseolin and major lectins. AB - BACKGROUND: A deficiency in phaseolin and phytohemagglutinin is associated with a near doubling of sulfur amino acid content in genetically related lines of common bean (Phaseolus vulgaris), particularly cysteine, elevated by 70%, and methionine, elevated by 10%. This mostly takes place at the expense of an abundant non-protein amino acid, S-methyl-cysteine. The deficiency in phaseolin and phytohemagglutinin is mainly compensated by increased levels of the 11S globulin legumin and residual lectins. Legumin, albumin-2, defensin and albumin-1 were previously identified as contributing to the increased sulfur amino acid content in the mutant line, on the basis of similarity to proteins from other legumes. RESULTS: Profiling of free amino acid in developing seeds of the BAT93 reference genotype revealed a biphasic accumulation of gamma-glutamyl-S-methyl cysteine, the main soluble form of S-methyl-cysteine, with a lag phase occurring during storage protein accumulation. A collection of 30,147 expressed sequence tags (ESTs) was generated from four developmental stages, corresponding to distinct phases of gamma-glutamyl-S-methyl-cysteine accumulation, and covering the transitions to reserve accumulation and dessication. Analysis of gene ontology categories indicated the occurrence of multiple sulfur metabolic pathways, including all enzymatic activities responsible for sulfate assimilation, de novo cysteine and methionine biosynthesis. Integration of genomic and proteomic data enabled the identification and isolation of cDNAs coding for legumin, albumin-2, defensin D1 and albumin-1A and -B induced in the absence of phaseolin and phytohemagglutinin. Their deduced amino acid sequences have a higher content of cysteine than methionine, providing an explanation for the preferential increase of cysteine in the mutant line. CONCLUSION: The EST collection provides a foundation to further investigate sulfur metabolism and the differential accumulation of sulfur amino acids in seed of common bean. Identification of sulfur-rich proteins whose levels are elevated in seed lacking phaseolin and phytohemagglutinin and sulfur metabolic genes may assist the improvement of protein quality. PMID- 21615928 TI - A graphical method for practical and informative identifiability analyses of physiological models: a case study of insulin kinetics and sensitivity. AB - BACKGROUND: Derivative based a-priori structural identifiability analyses of mathematical models can offer valuable insight into the identifiability of model parameters. However, these analyses are only capable of a binary confirmation of the mathematical distinction of parameters and a positive outcome can begin to lose relevance when measurement error is introduced. This article presents an integral based method that allows the observation of the identifiability of models with two-parameters in the presence of assay error. METHODS: The method measures the distinction of the integral formulations of the parameter coefficients at the proposed sampling times. It can thus predict the susceptibility of the parameters to the effects of measurement error. The method is tested in-silico with Monte Carlo analyses of a number of insulin sensitivity test applications. RESULTS: The method successfully captured the analogous nature of identifiability observed in Monte Carlo analyses of a number of cases including protocol alterations, parameter changes and differences in participant behaviour. However, due to the numerical nature of the analyses, prediction was not perfect in all cases. CONCLUSIONS: Thus although the current method has valuable and significant capabilities in terms of study or test protocol design, additional developments would further strengthen the predictive capability of the method. Finally, the method captures the experimental reality that sampling error and timing can negate assumed parameter identifiability and that identifiability is a continuous rather than discrete phenomenon. PMID- 21615930 TI - Patient medical costs for tuberculosis treatment and impact on adherence in China: a systematic review. AB - BACKGROUND: Charging for tuberculosis (TB) treatment could reduce completion rates, particularly in the poor. We identified and synthesised studies that measure costs of TB treatment, estimates of adherence and the potential impact of charging on treatment completion in China. METHODS: Inclusion criteria were primary research studies, including surveys and studies using qualitative methods, conducted in mainland China. We searched MEDLINE, PUBMED, EMBASE, Science Direct, HEED, CNKI to June 2010; and web pages of relevant Chinese and international organisations. Cost estimates were extracted, transformed, and expressed in absolute values and as a percentage of household income. RESULTS: Low income patients, defined at household or district level, pay a total of US$ 149 to 724 (RMB 1241 to 5228) for medical costs for a treatment course; as a percentage of annual household income, estimates range from 42% to 119%. One national survey showed 73% of TB patients at the time of the survey had interrupted or suspended treatment, and estimates from 9 smaller more recent studies showed that the proportion of patients at the time of the survey who had run out of drugs or were not taking them ranged from 3 to 25%. Synthesis of surveys and qualitative research indicate that cost is the most cited reason for default. CONCLUSIONS: Despite a policy of free drug treatment for TB in China, health services charge all income groups, and costs are high. Adherence measured in cross sectional surveys is often low, and the cumulative failure to adhere is likely to be much higher. These findings may be relevant to those concerned with the development and spread of multi-drug resistant TB. New strategies need to take this into account and ensure patient adherence. PMID- 21615929 TI - Dexamethasone inhibits the Nox-dependent ROS production via suppression of MKP-1 dependent MAPK pathways in activated microglia. AB - BACKGROUND: Nox-2 (also known as gp91phox), a subunit component of NADPH oxidases, generates reactive oxygen species (ROS). Nox-dependent ROS generation and nitric oxide (NO) release by microglia have been implicated in a variety of diseases in the central nervous system. Dexamethasone (Dex) has been shown to suppress the ROS production, NO release and inflammatory reaction of activated microglial cells. However, the underlying mechanisms remain unclear. RESULTS: The present study showed that the increased ROS production and NO release in activated BV-2 microglial cells by LPS were associated with increased expression of Nox-2 and iNOS. Dex suppressed the upregulation of Nox-2 and iNOS, as well as the subsequent ROS production and NO synthesis in activated BV-2 cells. This inhibition caused by Dex appeared to be mediated by upregulation of MAPK phosphatase-1 (MKP-1), which antagonizes the activity of mitogen-activated protein kinases (MAPKs). Dex induced-suppression of Nox-2 and -upregulation of MKP-1 was also evident in the activated microglia from corpus callosum of postnatal rat brains. The overexpression of MKP-1 or inhibition of MAPKs (by specific inhibitors of JNK and p38 MAPKs), were found to downregulate the expression of Nox-2 and iNOS and thereby inhibit the synthesis of ROS and NO in activated BV-2 cells. Moreover, Dex was unable to suppress the LPS-induced synthesis of ROS and NO in BV-2 cells transfected with MKP-1 siRNA. On the other hand, knockdown of Nox-2 in BV-2 cells suppressed the LPS-induced ROS production and NO release. CONCLUSION: In conclusion, it is suggested that downregulation of Nox-2 and overexpression of MKP-1 that regulate ROS and NO may form the potential therapeutic strategy for the treatment of neuroinflammation in neurodegenerative diseases. PMID- 21615931 TI - Selection of reference genes for normalization of quantitative real-time PCR in organ culture of the rat and rabbit intervertebral disc. AB - BACKGROUND: The accuracy of quantitative real-time RT-PCR (qRT-PCR) is often influenced by experimental artifacts, resulting in erroneous expression profiles of target genes. The practice of employing normalization using a reference gene significantly improves reliability and its applicability to molecular biology. However, selection of an ideal reference gene(s) is of critical importance to discern meaningful results. The aim of this study was to evaluate the stability of seven potential reference genes (Actb, GAPDH, 18S rRNA, CycA, Hprt1, Ywhaz, and Pgk1) and identify most stable gene(s) for application in tissue culture research using the rat and rabbit intervertebral disc (IVD). FINDINGS: In vitro, four genes (Hprt1, CycA, GAPDH, and 18S rRNA) in rat IVD tissue and five genes (CycA, Hprt1, Actb, Pgk1, and Ywhaz) in rabbit IVD tissue were determined as most stable for up to 14 days in culture. Pair-wise variation analysis indicated that combination of Hprt1 and CycA in rat and the combination of Hprt1, CycA, and Actb in rabbit may most stable reference gene candidates for IVD tissue culture. CONCLUSIONS: Our results indicate that Hprt1 and CycA are the most stable reference gene candidates for rat and rabbit IVD culture studies. In rabbit IVD, Actb could be an additional gene employed in conjunction with Hprt1 and CycA. Selection of optimal reference gene candidate(s) should be a pertinent exercise before employment of PCR outcome measures for biomedical research. PMID- 21615932 TI - Challenges of controlling sleeping sickness in areas of violent conflict: experience in the Democratic Republic of Congo. AB - BACKGROUND: Human African trypanosomiasis (HAT), or sleeping sickness, is a fatal neglected tropical disease if left untreated. HAT primarily affects people living in rural sub-Saharan Africa, often in regions afflicted by violent conflict. Screening and treatment of HAT is complex and resource-intensive, and especially difficult in insecure, resource-constrained settings. The country with the highest endemicity of HAT is the Democratic Republic of Congo (DRC), which has a number of foci of high disease prevalence. We present here the challenges of carrying out HAT control programmes in general and in a conflict-affected region of DRC. We discuss the difficulties of measuring disease burden, medical care complexities, waning international support, and research and development barriers for HAT. DISCUSSION: In 2007, Medecins Sans Frontieres (MSF) began screening for HAT in the Haut-Uele and Bas-Uele districts of Orientale Province in northeastern DRC, an area of high prevalence affected by armed conflict. Through early 2009, HAT prevalence rate of 3.4% was found, reaching 10% in some villages. More than 46,000 patients were screened and 1,570 treated for HAT during this time. In March 2009, two treatment centres were forced to close due to insecurity, disrupting patient treatment, follow-up, and transmission-control efforts. One project was reopened in December 2009 when the security situation improved, and another in late 2010 based on concerns that population displacement might reactivate historic foci. In all of 2010, 770 patients were treated at these sites, despite a limited geographical range of action for the mobile teams. SUMMARY: In conflict settings where HAT is prevalent, targeted medical interventions are needed to provide care to the patients caught in these areas. Strategies of integrating care into existing health systems may be unfeasible since such infrastructure is often absent in resource-poor contexts. HAT care in conflict areas must balance logistical and medical capacity with security considerations, and community networks and international-response coordination should be maintained. Research and development for less complicated, field adapted tools for diagnosis and treatment, and international support for funding and program implementation, are urgently needed to facilitate HAT control in these remote and insecure areas. PMID- 21615934 TI - Screening and brief interventions for hazardous and harmful alcohol use among patients with active tuberculosis attending primary care clinics in South Africa: a cluster randomized controlled trial protocol. AB - BACKGROUND: In 2008 the World Health Organization (WHO) reported that South Africa had the highest tuberculosis (TB) incidence in the world. This high incidence rate is linked to a number of factors, including HIV co-infection and alcohol use disorders. The diagnosis and treatment package for TB and HIV co infection is relatively well established in South Africa. However, because alcohol use disorders may present more insidiously, making it difficult to diagnose, those patients with active TB and misusing alcohol are not easily cured from TB. With this in mind, the primary purpose of this cluster randomized controlled trial is to provide screening for alcohol misuse and to test the efficacy of brief interventions in reducing alcohol intake in those patients with active TB found to be misusing alcohol in primary health care clinics in three provinces in South Africa. METHODS/DESIGN: Within each of the three selected health districts with the highest TB burden in South Africa, 14 primary health care clinics with the highest TB caseloads will be selected. Those agreeing to participate will be stratified according to TB treatment caseload and the type of facility (clinic or community health centre). Within strata from 14 primary care facilities, 7 will be randomly selected into intervention and 7 to control study clinics (42 clinics, 21 intervention clinics and 21 control clinics). At the clinic level systematic sampling will be used to recruit newly diagnosed TB patients. Those consenting will be screened for alcohol misuse using the AUDIT. Patients who screen positive for alcohol misuse over a 6-month period will be given either a brief intervention based on the Information-Motivation-Behavioural Skills (IMB) Model or an alcohol use health education leaflet.A total sample size of 520 is expected. DISCUSSION: The trial will evaluate the impact of alcohol screening and brief interventions for patients with active TB in primary care settings in South Africa. The findings will impact public health and will enable the health ministry to formulate policy related to comprehensive treatment for TB and alcohol misuse, which will result in reduction in alcohol use and ultimately improve the TB cure rates. TRIAL REGISTRATION NUMBER: PACTR: PACTR201105000297151. PMID- 21615933 TI - Signal peptide cleavage is essential for surface expression of a regulatory T cell surface protein, leucine rich repeat containing 32 (LRRC32). AB - BACKGROUND: Elevated numbers of regulatory T cells (T(regs)) have been implicated in certain cancers. Depletion of T(regs) has been shown to increase anti-tumor immunity. T(regs) also play a critical role in the suppression of autoimmune responses. The study of T(regs) has been hampered by a lack of adequate surface markers. Leucine Rich Repeat Containing 32 (LRRC32), also known as Glycoprotein A Repetitions Predominant (GARP), has been postulated as a novel surface marker of activated T(regs). However, there is limited information regarding the processing of LRRC32 or the regulatory phenotype and functional activity of T(regs) expressing LRRC32. RESULTS: Using naturally-occurring freshly isolated T(regs), we demonstrate that low levels of LRRC32 are present intracellularly prior to activation and that freshly isolated LRRC32+ T(regs) are distinct from LRRC32- T(regs) with respect to the expression of surface CD62L. Using LRRC32 transfectants of HEK cells, we demonstrate that the N-terminus of LRRC32 is cleaved prior to expression of the protein at the cell surface. Furthermore, we demonstrate using a construct containing a deleted putative signal peptide region that the presence of a signal peptide region is critical to cell surface expression of LRRC32. Finally, mixed lymphocyte assays demonstrate that LRRC32+ T(regs) are more potent suppressors than LRRC32- T(regs). CONCLUSIONS: A cleaved signal peptide site in LRRC32 is necessary for surface localization of native LRRC32 following activation of naturally-occurring freshly-isolated regulatory T cells. LRRC32 expression appears to alter the surface expression of activation markers of T cells such as CD62L. LRRC32 surface expression may be useful as a marker that selects for more potent T(reg) populations. In summary, understanding the processing and expression of LRRC32 may provide insight into the mechanism of action of T(regs) and the refinement of immunotherapeutic strategies aimed at targeting these cells. PMID- 21615935 TI - Synchronous gastric adenocarcinoma and gastrointestinal stromal tumor (GIST) of the stomach: a case report. AB - Gastrointestinal stromal tumors (GISTs) are rare mesenchymal neoplasms of the gastrointestinal tract (1%), and stomach is the most common location involved. However, the co-existence of gastric adenocarcinoma and GIST is very rare. A case of an 80-year-old male with a simultaneous presentation of a gastric adenocarcinoma and GIST is presented. Various hypotheses have been proposed in order to explain this rare simultaneous development, but even though it's cause has not been proven yet. PMID- 21615936 TI - Designing clinical trials for assessing the effects of cognitive training and physical activity interventions on cognitive outcomes: the Seniors Health and Activity Research Program Pilot (SHARP-P) study, a randomized controlled trial. AB - BACKGROUND: The efficacy of non-pharmacological intervention approaches such as physical activity, strength, and cognitive training for improving brain health has not been established. Before definitive trials are mounted, important design questions on participation/adherence, training and interventions effects must be answered to more fully inform a full-scale trial. METHODS: SHARP-P was a single blinded randomized controlled pilot trial of a 4-month physical activity training intervention (PA) and/or cognitive training intervention (CT) in a 2 * 2 factorial design with a health education control condition in 73 community dwelling persons, aged 70-85 years, who were at risk for cognitive decline but did not have mild cognitive impairment. RESULTS: Intervention attendance rates were higher in the CT and PACT groups: CT: 96%, PA: 76%, PACT: 90% (p=0.004), the interventions produced marked changes in cognitive and physical performance measures (p<=0.05), and retention rates exceeded 90%. There were no statistically significant differences in 4-month changes in composite scores of cognitive, executive, and episodic memory function among arms. Four-month improvements in the composite measure increased with age among participants assigned to physical activity training but decreased with age for other participants (intervention*age interaction p=0.01). Depending on the choice of outcome, two-armed full-scale trials may require fewer than 1,000 participants (continuous outcome) or 2,000 participants (categorical outcome). CONCLUSIONS: Good levels of participation, adherence, and retention appear to be achievable for participants through age 85 years. Care should be taken to ensure that an attention control condition does not attenuate intervention effects. Depending on the choice of outcome measures, the necessary sample sizes to conduct four-year trials appear to be feasible. TRIAL REGISTRATION: Clinicaltrials.gov Identifier: NCT00688155. PMID- 21615937 TI - A synthetic system links FeFe-hydrogenases to essential E. coli sulfur metabolism. AB - BACKGROUND: FeFe-hydrogenases are the most active class of H2-producing enzymes known in nature and may have important applications in clean H2 energy production. Many potential uses are currently complicated by a crucial weakness: the active sites of all known FeFe-hydrogenases are irreversibly inactivated by O2. RESULTS: We have developed a synthetic metabolic pathway in E. coli that links FeFe-hydrogenase activity to the production of the essential amino acid cysteine. Our design includes a complementary host strain whose endogenous redox pool is insulated from the synthetic metabolic pathway. Host viability on a selective medium requires hydrogenase expression, and moderate O2 levels eliminate growth. This pathway forms the basis for a genetic selection for O2 tolerance. Genetically selected hydrogenases did not show improved stability in O2 and in many cases had lost H2 production activity. The isolated mutations cluster significantly on charged surface residues, suggesting the evolution of binding surfaces that may accelerate hydrogenase electron transfer. CONCLUSIONS: Rational design can optimize a fully heterologous three-component pathway to provide an essential metabolic flux while remaining insulated from the endogenous redox pool. We have developed a number of convenient in vivo assays to aid in the engineering of synthetic H2 metabolism. Our results also indicate a H2 independent redox activity in three different FeFe-hydrogenases, with implications for the future directed evolution of H2-activating catalysts. PMID- 21615938 TI - Genetic polymorphisms in folate pathway enzymes, DRD4 and GSTM1 are related to temporomandibular disorder. AB - BACKGROUND: Temporomandibular disorder (TMD) is a multifactorial syndrome related to a critical period of human life. TMD has been associated with psychological dysfunctions, oxidative state and sexual dimorphism with coincidental occurrence along the pubertal development. In this work we study the association between TMD and genetic polymorphisms of folate metabolism, neurotransmission, oxidative and hormonal metabolism. Folate metabolism, which depends on genes variations and diet, is directly involved in genetic and epigenetic variations that can influence the changes of last growing period of development in human and the appearance of the TMD. METHODS: A case-control study was designed to evaluate the impact of genetic polymorphisms above described on TMD. A total of 229 individuals (69% women) were included at the study; 86 were patients with TMD and 143 were healthy control subjects. Subjects underwent to a clinical examination following the guidelines by the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD). Genotyping of 20 Single Nucleotide Polymorphisms (SNPs), divided in two groups, was performed by multiplex minisequencing preceded by multiplex PCR. Other seven genetic polymorphisms different from SNPs (deletions, insertions, tandem repeat, null genotype) were achieved by a multiplex-PCR. A chi-square test was performed to determine the differences in genotype and allelic frequencies between TMD patients and healthy subjects. To estimate TMD risk, in those polymorphisms that shown significant differences, odds ratio (OR) with a 95% of confidence interval were calculated. RESULTS: Six of the polymorphisms showed statistical associations with TMD. Four of them are related to enzymes of folates metabolism: Allele G of Serine Hydoxymethyltransferase 1 (SHMT1) rs1979277 (OR = 3.99; 95%CI 1.72, 9.25; p = 0.002), allele G of SHMT1 rs638416 (OR = 2.80; 95%CI 1.51, 5.21; p = 0.013), allele T of Methylentetrahydrofolate Dehydrogenase (MTHFD) rs2236225 (OR = 3.09; 95%CI 1.27, 7.50; p = 0.016) and allele A of Methionine Synthase Reductase (MTRR) rs1801394 (OR = 2.35; 95CI 1.10, 5.00; p = 0.037). An inflammatory oxidative stress enzyme, Gluthatione S-Tranferase Mu-1(GSTM1), null allele (OR = 2.21; 95%CI 1.24, 4.36; p = 0.030) and a neurotransmission receptor, Dopamine Receptor D4 (DRD4), long allele of 48 bp-repeat (OR = 3.62; 95%CI 0.76, 17.26; p = 0.161). CONCLUSIONS: Some genetic polymorphisms related to folates metabolism, inflammatory oxidative stress, and neurotransmission responses to pain, has been significantly associated to TMD syndrome. PMID- 21615939 TI - Cement leakage causes potential thermal injury in vertebroplasty. AB - BACKGROUND: Percutaneous vertebroplasty by injecting PMMA bone cement into the fractured vertebrae has been widely accepted in treatment of spinal compression fracture. However, the exothermic polymerization of bone cement may cause osseous or neural tissue injury. This study is thus designed to evaluate the potential risk of thermal damage in percutaneous vertebroplasty. METHOD: Twelve porcine vertebrae were immersed in 37 degrees C saline for the experiment. In the first stage of the study, vertebroplasty without cement leakage (control group, n = 6) was simulated. The anterior cortex, foramen, posterior cortex and the center of the vertebral body were selected for temperature measurement. Parameters including peak temperature and duration above 45 degrees C were recorded. In the second stage, a model (n = 6) simulating bone cement leaking into the spinal canal was designed. The methods for temperature measurement were identical to those used in the first stage. RESULTS: In Stage 1 of the study (vertebroplasty of the porcine vertebral body in the absence of cement leakage), the average maximal temperature at the anterior cortex was 42.4 +/- 2.2 degrees C; at the neural foramen 39.5 +/- 2.1 degrees C; at the posterior cortex 40.0 +/- 2.5 degrees C and at the vertebral center, 68.1 +/- 3.4 degrees C. The average time interval above 45 degrees C was 0 seconds at the anterior cortex; at the neural foramen, 0 seconds; at the posterior cortex, 0 seconds and at the vertebral center, 223 seconds. Thus, except at the core of the bone cement, temperatures around the vertebral body did not exceed 45 degrees C. In Stage 2 of the study (cement leakage model), the average maximal temperature at the anterior cortex was 42.7 +/- 2.4 degrees C; at the neural foramen, 41.1 +/- 0.4 degrees C; at the posterior cortex, 59.1 +/- 7.6 degrees C and at the vertebral center, 77.3 +/- 5.7 degrees C. The average time interval above 45 degrees C at the anterior cortex was 0 seconds; at the neural foramen, 0 seconds; at the posterior cortex, 329.3 seconds and at the vertebral center, 393.2 seconds. Based on these results, temperatures exceeded 45 degrees C at the posterior cortex and at the vertebral center. CONCLUSIONS: The results indicated that, for bone cement confined within the vertebra, curing temperatures do not directly cause thermal injury to the nearby soft tissue. If bone cement leaks into the spinal canal, the exothermic reaction at the posterior cortex might result in thermal injury to the neural tissue. PMID- 21615941 TI - Fast empirical Bayesian LASSO for multiple quantitative trait locus mapping. AB - BACKGROUND: The Bayesian shrinkage technique has been applied to multiple quantitative trait loci (QTLs) mapping to estimate the genetic effects of QTLs on quantitative traits from a very large set of possible effects including the main and epistatic effects of QTLs. Although the recently developed empirical Bayes (EB) method significantly reduced computation comparing with the fully Bayesian approach, its speed and accuracy are limited by the fact that numerical optimization is required to estimate the variance components in the QTL model. RESULTS: We developed a fast empirical Bayesian LASSO (EBLASSO) method for multiple QTL mapping. The fact that the EBLASSO can estimate the variance components in a closed form along with other algorithmic techniques render the EBLASSO method more efficient and accurate. Comparing with the EB method, our simulation study demonstrated that the EBLASSO method could substantially improve the computational speed and detect more QTL effects without increasing the false positive rate. Particularly, the EBLASSO algorithm running on a personal computer could easily handle a linear QTL model with more than 100,000 variables in our simulation study. Real data analysis also demonstrated that the EBLASSO method detected more reasonable effects than the EB method. Comparing with the LASSO, our simulation showed that the current version of the EBLASSO implemented in Matlab had similar speed as the LASSO implemented in Fortran, and that the EBLASSO detected the same number of true effects as the LASSO but a much smaller number of false positive effects. CONCLUSIONS: The EBLASSO method can handle a large number of effects possibly including both the main and epistatic QTL effects, environmental effects and the effects of gene-environment interactions. It will be a very useful tool for multiple QTL mapping. PMID- 21615942 TI - Idiopathic pancreatitis is a consequence of an altering spectrum of bile nucleation time. AB - BACKGROUND: The pathogenesis of idiopathic pancreatitis (IP) remains poorly understood. Our hypothesis is that IP is a sequel of micro-crystallization of hepatic bile. METHODS: A prospective case control study compared 55 patients; symptomatic cholelithiasis - 30 (14 male, median age 36 years; mean BMI - 25.1 kg/m2), gallstone pancreatitis - 9 (3 male, median age 35 years; mean BMI - 24.86 kg/m2 ) and IP - 16 (9 male, median age 34 years; mean BMI -23.34 kg/m2) with 30 controls (15 male, median age 38 years; mean BMI = 24.5 kg/m2) undergoing laparotomy for conditions not related to the gall bladder and bile duct. Ultrafiltered bile from the common hepatic duct in patients and controls was incubated in anaerobic conditions and examined by polarized light microscopy to assess bile nucleation time (NT). In the analysis, the mean NT of patients with gallstones and gallstone pancreatitis was taken as a cumulative mean NT for those with established gallstone disease (EGD). RESULTS: Patients were similar to controls. Mean NT in all groups of patients was significantly shorter than controls (EGD cumulative mean NT, 1.73 +/- 0.2 days vs. controls, 12.74 +/- 0.4 days, P = 0.001 and IP patients mean NT, 3.1 +/- 0.24 days vs. controls, 12.74 +/ 0.4 days, P = 0.001). However, NT in those with IP was longer compared with those with EGD (mean NT in IP, 3.1 +/- 0.24 days vs. cumulative mean in EGD: 1.73 +/- 0.2 days, P = 0.002). CONCLUSION: Nucleation time of bile in patients with IP is abnormal and is intermediate to nucleation time of lithogenic bile at one end of the spectrum of lithogenicity and non-lithogenic bile, at the other end. PMID- 21615940 TI - Are workplace health promotion programs effective at improving presenteeism in workers? A systematic review and best evidence synthesis of the literature. AB - BACKGROUND: Presenteeism is highly prevalent and costly to employers. It is defined as being present at work, but limited in some aspect of job performance by a health problem.Workplace health promotion (WHP) is a common strategy used to enhance on-the-job productivity. The primary objective is to determine if WHP programs are effective in improving presenteeism. The secondary objectives are to identify characteristics of successful programs and potential risk factors for presenteeism. METHODS: The Cochrane Library, Medline, and other electronic databases were searched from 1990 to 2010. Reference lists were examined, key journals were hand-searched and experts were contacted. Included studies were original research that contained data on at least 20 participants (>= 18 years of age), and examined the impacts of WHP programs implemented at the workplace. The Effective Public Health Practice Project Tool for Quantitative Studies was used to rate studies. 'Strong' and 'moderate' studies were abstracted into evidence tables, and a best evidence synthesis was performed. Interventions were deemed successful if they improved the outcome of interest. Their program components were identified, as were possible risk factors contributing to presenteeism. RESULTS: After 2,032 titles and abstracts were screened, 47 articles were reviewed, and 14 were accepted (4 strong and 10 moderate studies). These studies contained preliminary evidence for a positive effect of some WHP programs. Successful programs offered organizational leadership, health risk screening, individually tailored programs, and a supportive workplace culture. Potential risk factors contributing to presenteeism included being overweight, a poor diet, a lack of exercise, high stress, and poor relations with co-workers and management. LIMITATIONS: This review is limited to English publications. A large number of reviewed studies (70%) were inadmissible due to issues of bias, thus limiting the amount of primary evidence. The uncertainties surrounding presenteeism measurement is of significant concern as a source of bias. CONCLUSIONS: The presenteeism literature is young and heterogeneous. There is preliminary evidence that some WHP programs can positively affect presenteeism and that certain risk factors are of importance. Future research would benefit from standard presenteeism metrics and studies conducted across a broad range of workplace settings. PMID- 21615943 TI - The profile of general practitioners (GPs) who publish in selected family practice journals. AB - BACKGROUND: Providing support for research is one of the key issues in the ongoing attempts to improve Primary Care. However, when patient care takes up a significant part of a GP's time, conducting research is difficult. In this study we examine the working conditions and profile of GPs who publish in three leading medical journals and propose possible remedial policy actions. FINDINGS: The authors of all articles published in 2006 and 2007 in three international Family Medicine journals - Annals of Family Medicine, Family Practice, and Journal of Family Practice - were contacted by E-mail. They were asked to complete a questionnaire investigating the following variables: availability of specific time for research, time devoted to research, number of patients attended, and university affiliation. Only GPs were included in the study. Three hundred and ten relevant articles published between 2006 and 2007 were identified and the authors contacted using a survey tool. 124 researchers responded to our questionnaire; 45% of respondents who were not GPs were excluded. On average GPs spent 2.52 days per week and 6.9 hours per day on patient care, seeing 45 patients per week. Seventy-five per cent of GPs had specific time assigned to research, on average 13 hours per week; 79% were affiliated to a university and 69% held teaching positions. CONCLUSIONS: Most GPs who publish original articles in leading journals have time specifically assigned to research as part of their normal working schedule. They see a relatively small number of patients. Improving the working conditions of family physicians who intend to investigate is likely to lead to better research results. PMID- 21615944 TI - Evaluation of allelic forms of the erythrocyte binding antigen 175 (EBA-175) in Plasmodium falciparum field isolates from Brazilian endemic area. AB - BACKGROUND: The Plasmodium falciparum Erythrocyte Binding Antigen-175 (EBA-175) is an antigen considered to be one of the leading malaria vaccine candidates. EBA 175 mediates sialic acid-dependent binding to glycophorin A on the erythrocytes playing a crucial role during invasion of the P. falciparum in the host cell. Dimorphic allele segments, termed C-fragment and F-fragment, have been found in high endemicity malaria areas and associations between the dimorphism and severe malaria have been described. In this study, the genetic dimorphism of EBA-175 was evaluated in P. falciparum field isolates from Brazilian malaria endemic area. METHODS: The study was carried out in rural villages situated near Porto Velho, Rondonia State in the Brazilian Amazon in three time points between 1993 and 2008. The allelic dimorphism of the EBA-175 was analysed by Nested PCR. RESULTS: The classical allelic dimorphism of the EBA-175 was identified in the studied area. Overall, C-fragment was amplified in a higher frequency than F-fragment. The same was observed in the three time points where C-fragment was observed in a higher frequency than F-fragment. Single infections (one fragment amplified) were more frequent than mixed infection (two fragments amplified). CONCLUSIONS: These findings confirm the dimorphism of EBA175, since only the two types of fragments were amplified, C-fragment and F-fragment. Also, the results show the remarkable predominance of CAMP allele in the studied area. The comparative analysis in three time points indicates that the allelic dimorphism of the EBA-175 is stable over time. PMID- 21615946 TI - Delay in diagnosis of tuberculosis in Rawalpindi, Pakistan. AB - BACKGROUND: Delay in diagnosis and treatment of tuberculosis (TB) may enhance the chances of morbidity and mortality and play a key role in continuous transmission of the bacilli. The objective of this study was to describe health care seeking behavior of suspected TB patients and initial diagnostic work up prior to consultation and diagnosis at National TB Center (NTC). FINDINGS: Interviews of 252 sputum smear positive patients were taken from NTC, Rawalpindi. The duration between on-set of symptoms and start of treatment was considered as the total delay and correlated with general characteristics of TB patients. The proportion of males and females were 49.6% and 50.4% with median age of 25 and 24 years respectively. A median delay of 56 days (8 weeks) was observed which was significantly associated with age, cough and fever. More than 50% of the current patients had a history of contact with previously diagnosed TB patients. The majority of patients (63%) visited health care providers within three weeks of appearance of symptoms but only thirty five percent were investigated for TB diagnosis. CONCLUSION: Cough and fever are being ignored as likely symptoms of TB by patients as well as health care providers resulting in delay. Engaging private practitioners through public private mix (PPM) approach for expansion of TB diagnosis and increasing public awareness could be more beneficial to reduce delay. PMID- 21615945 TI - Low penetrance of retinoblastoma for p.V654L mutation of the RB1 gene. AB - BACKGROUND: Retinoblastoma is caused by compound heterozygosity or homozygosity of retinoblastoma gene (RB1) mutations. In germline retinoblastoma, mutations in the RB1 gene predispose individuals to increased cancer risks during development. These mutations segregate as autosomal dominant traits with high penetrance (90%). METHODS: We screened 30 family members from one family using high resolution melting assay and DNA direct sequencing for mutations in the RB1 gene. We evaluate the phenotype and penetrance of germline mutations of the RB1 gene in a large Taiwanese family. RESULTS: The molecular analysis and clinical details of this family showed phenotypic variability associated with the p.V654L mutation in exon 19 of the RB1 gene in 11 family members. The phenotype varied from asymptomatic to presence of a unilateral tumor. Only four individuals (2 males and 2 females) developed unilateral retinoblastoma, which resulted in calculated low penetrance of 36% (4/11). The four individuals with retinoblastoma were diagnosed before the age of three years. None of their relatives exhibited variable severity or bilateral retinoblastoma. CONCLUSIONS: The diseased-eye ratio for this family was 0.36, which is lower than current estimates. This suggests that the RB1 p.V654L mutation is a typical mutation associated with low penetrance. PMID- 21615947 TI - Age-specific mortality patterns in Central Mozambique during and after the end of the Civil War. AB - BACKGROUND: In recent years, vigorous debate has developed concerning how conflicts contribute to the spread of infectious diseases, and in particular, the role of post-conflict situations in the epidemiology of HIV/AIDS. This study details the age-specific mortality patterns among the population in the central provincial capital of Beira, Mozambique, during and after the Mozambican civil war which ended in 1992. METHODS: Data was collected from the death register at Beira's Central Hospital between 1985 and 2003 and descriptively analyzed. RESULTS: The data show two distinct periods: before and after the peace agreements in 1992. Before 1992 (during the civil war), the main impact of mortality was on children below 5 years of age, including still births, accounting for 58% of all deaths. After the war ended in 1992, the pattern shifted dramatically and rapidly to the 15-49 year old age group which accounted for 49% of all deaths by 2003. CONCLUSIONS: As under-5 mortality rates were decreasing at the end of the conflict, rates for 24-49 year old adults began to dramatically increase due to AIDS. This study demonstrates that strategies can be implemented during conflicts to decrease mortality rates in one vulnerable population but post-conflict dynamics can bring together other factors which contribute to the rapid spread of other infectious diseases in other vulnerable populations. PMID- 21615948 TI - Assessing implementation difficulties in tobacco use prevention and cessation counselling among dental providers. AB - BACKGROUND: Tobacco use adversely affects oral health. Clinical guidelines recommend that dental providers promote tobacco abstinence and provide patients who use tobacco with brief tobacco use cessation counselling. Research shows that these guidelines are seldom implemented, however. To improve guideline adherence and to develop effective interventions, it is essential to understand provider behaviour and challenges to implementation. This study aimed to develop a theoretically informed measure for assessing among dental providers implementation difficulties related to tobacco use prevention and cessation (TUPAC) counselling guidelines, to evaluate those difficulties among a sample of dental providers, and to investigate a possible underlying structure of applied theoretical domains. METHODS: A 35-item questionnaire was developed based on key theoretical domains relevant to the implementation behaviours of healthcare providers. Specific items were drawn mostly from the literature on TUPAC counselling studies of healthcare providers. The data were collected from dentists (n = 73) and dental hygienists (n = 22) in 36 dental clinics in Finland using a web-based survey. Of 95 providers, 73 participated (76.8%). We used Cronbach's alpha to ascertain the internal consistency of the questionnaire. Mean domain scores were calculated to assess different aspects of implementation difficulties and exploratory factor analysis to assess the theoretical domain structure. The authors agreed on the labels assigned to the factors on the basis of their component domains and the broader behavioural and theoretical literature. RESULTS: Internal consistency values for theoretical domains varied from 0.50 ('emotion') to 0.71 ('environmental context and resources'). The domain environmental context and resources had the lowest mean score (21.3%; 95% confidence interval [CI], 17.2 to 25.4) and was identified as a potential implementation difficulty. The domain emotion provided the highest mean score (60%; 95% CI, 55.0 to 65.0). Three factors were extracted that explain 70.8% of the variance: motivation (47.6% of variance, alpha = 0.86), capability (13.3% of variance, alpha = 0.83), and opportunity (10.0% of variance, alpha = 0.71). CONCLUSIONS: This study demonstrated a theoretically informed approach to identifying possible implementation difficulties in TUPAC counselling among dental providers. This approach provides a method for moving from diagnosing implementation difficulties to designing and evaluating interventions. PMID- 21615949 TI - Adipokine resistin predicts anti-inflammatory effect of glucocorticoids in asthma. AB - BACKGROUND: Adipokines are protein mediators secreted by adipose tissue. Recently, adipokines have also been involved in the regulation of inflammation and allergic responses, and suggested to affect the risk of asthma especially in obese female patients. We assessed if adipokines predict responsiveness to glucocorticoids and if plasma adipokine levels are associated with lung function or inflammatory activity also in non-obese (body mass index (BMI) <= 30 kg/m2) women with newly-diagnosed steroid-naive asthma. METHODS: Lung function, exhaled NO, plasma levels of adipokines leptin, resistin, adiponectin and adipsin, and inflammatory markers were measured in 35 steroid-naive female asthmatics and in healthy controls. The measurements were repeated in a subgroup of asthmatics after 8 weeks of treatment with inhaled fluticasone. Adipokine concentrations in plasma were adjusted for BMI. RESULTS: High baseline resistin concentrations were associated with a more pronounced decrease in serum levels of eosinophil cationic protein (ECP) (r = -0.745, p = 0.013), eosinophil protein X (EPX) (r = -0.733, p = 0.016) and myeloperoxidase (MPO) (r = -0.721, p = 0.019) during fluticasone treatment. In asthmatics, leptin correlated positively with asthma symptom score and negatively with lung function. However, no significant differences in plasma adipokine levels between non-obese asthmatics and healthy controls were found. The effects of resistin were also investigated in human macrophages in cell culture. Interestingly, resistin increased the production of proinflammatory factors IL-6 and TNF-alpha and that was inhibited by fluticasone. CONCLUSIONS: High resistin levels predicted favourable anti-inflammatory effect of inhaled glucocorticoids suggesting that resistin may be a marker of steroid-sensitive phenotype in asthma. High leptin levels were associated with a more severe disease suggesting that the link between leptin and asthma is not restricted to obesity. PMID- 21615950 TI - Histone deacetylase inhibitors SAHA and sodium butyrate block G1-to-S cell cycle progression in neurosphere formation by adult subventricular cells. AB - BACKGROUND: Histone deacetylases (HDACs) are enzymes that modulate gene expression and cellular processes by deacetylating histones and non-histone proteins. While small molecule inhibitors of HDAC activity (HDACi) are used clinically in the treatment of cancer, pre-clinical treatment models suggest they also exert neuroprotective effects and stimulate neurogenesis in neuropathological conditions. However, the direct effects of HDACi on cell cycle progression and proliferation, two properties required for continued neurogenesis, have not been fully characterized in adult neural stem cells (NSCs). In this study, we examined the effects of two broad class I and class II HDACi on adult mouse NSCs, the hydroxamate-based HDACi suberoylanilide hydroxamic acid (vorinostat, SAHA) and the short chain fatty acid HDACi sodium butyrate. RESULTS: We show that both HDACi suppress the formation of neurospheres by adult mouse NSCs grown in proliferation culture conditions in vitro. DNA synthesis is significantly inhibited in adult mouse NSCs exposed to either SAHA or sodium butyrate and inhibition is associated with an arrest in the G1 phase of the cell cycle. HDACi exposure also resulted in transcriptional changes in adult mouse NSCs. Cdk inhibitor genes p21 and p27 transcript levels are increased and associated with elevated H3K9 acetylation levels at proximal promoter regions of p21 and p27. mRNA levels for notch effector Hes genes and Spry-box stem cell transcription factors are downregulated, whereas pro-neural transcription factors Neurog1 and Neurod1 are upregulated. Lastly, we show HDAC inhibition under proliferation culture conditions leads to long-term changes in cell fate in adult mouse NSCs induced to differentiate in vitro. CONCLUSION: SAHA and sodium butyrate directly regulate cdk inhibitor transcription to control cell cycle progression in adult mouse NSCs. HDAC inhibition results in G1 arrest in adult mouse NSCs and transcriptional changes associated with activation of neuronal lineage commitment programs and a reduction of stem/progenitor state. Changes in differentiated cell state in adult mouse NSCs treated with HDACi under proliferation culture conditions suggests an intrinsic relationship between multipotency, cell cycle progression and HDAC activity in these cells. PMID- 21615951 TI - The ACCESS study a Zelen randomised controlled trial of a treatment package including problem solving therapy compared to treatment as usual in people who present to hospital after self-harm: study protocol for a randomised controlled trial. AB - BACKGROUND: People who present to hospital after intentionally harming themselves pose a common and important problem. Previous reviews of interventions have been inconclusive as existing trials have been under powered and done on unrepresentative populations. These reviews have however indicated that problem solving therapy and regular written communications after the self-harm attempt may be an effective treatment. This protocol describes a large pragmatic trial of a package of measures which include problem solving therapy, regular written communication, patient support, cultural assessment, improved access to primary care and a risk management strategy in people who present to hospital after self harm using a novel design. METHODS: We propose to use a double consent Zelen design where participants are randomised prior to giving consent to enroll a large representative cohort of patients. The main outcome will be hospital attendance following repetition of self-harm, in the 12 months after recruitment with secondary outcomes of self reported self-harm, hopelessness, anxiety, depression, quality of life, social function and hospital use at three months and one year. DISCUSSION: A strength of the study is that it is a pragmatic trial which aims to recruit large numbers and does not exclude people if English is not their first language. A potential limitation is the analysis of the results which is complex and may underestimate any effect if a large number of people refuse their consent in the group randomised to problem solving therapy as they will effectively cross over to the treatment as usual group. However the primary analysis is a true intention to treat analysis of everyone randomised which includes both those who consent and do not consent to participate in the study. This provides information about how the intervention will work in practice in a representative population which is a major advance in this study compared to what has been done before. TRIAL REGISTRATION: Australia and New Zealand Clinical Trials Register (ANZCTR): ACTRN12609000641291. PMID- 21615952 TI - Closure of post-laryngectomy pharyngocutaneous fistulae. AB - BACKGROUND: Closure of salvage laryngectomy defects with vascularized tissue remains controversial. METHODS: We evaluate outcomes in patients who required repair of a fistula after attempted primary closure of salvage laryngectomy defect and assess risk factors for persistent fistula. Between 2001 and 2010, 20 patients were treated for pharyngocutaneous fistulae after primary closure of a salvage laryngectomy. All patients required free flap repair for definitive fistula management. RESULTS: Patients presented with fistulae from one to 18 months in duration; median time to closure was seven days. Radial forearm free flap was used in 86% of patients. With free flap alone 50% of patients achieved fistula closure. Additional procedures improved closure rate to 85%. Recipient vessels were used in the neck in 54.5%, compared to internal mammary vessels in 45.5%. Hypothyroidism was identified as a risk factor for persistent fistula (p = 0.01). Chronic steroid use (p = 0.08) did not reach significance as a risk factor for fistula closure. Gastroesophageal reflux disease was newly diagnosed or noted as a comorbidity in 14 patients (70%) in this study. It did not reach statistical significance as a risk factor in refistulization (p = 0.12). Complications included leak, carotid blowout, infection, free flap loss, and late refistulization. Overall flap failure in this study was 4.5%. CONCLUSIONS: Delayed secondary repair of pharygocutaneous fistulas after salvage laryngectomy is associated with a higher complication rate and poor success rates compared to use of vascularized tissue at the time of salvage laryngectomy. Prolonged wound healing in these patients is associated with hypothyroidism. PMID- 21615953 TI - Relevance of circulating nucleosomes and oncological biomarkers for predicting response to transarterial chemoembolization therapy in liver cancer patients. AB - BACKGROUND: Transarterial chemoembolization (TACE) therapy is an effective locoregional treatment in hepatocellular cancer (HCC) patients. For early modification of therapy, markers predicting therapy response are urgently required. METHODS: Here, sera of 50 prospectively and consecutively included HCC patients undergoing 71 TACE therapies were taken before and 3 h, 6 h and 24 h after TACE application to analyze concentrations of circulating nucleosomes, cytokeratin-19 fragments (CYFRA 21-1), alpha fetoprotein (AFP), C-reactive protein (CRP) and several liver biomarkers, and to compare these with radiological response to therapy. RESULTS: While nucleosomes, CYFRA 21-1, CRP and some liver biomarkers increased already 24 h after TACE, percental changes of nucleosome concentrations before and 24 h after TACE and pre- and posttherapeutic values of AFP, gamma-glutamyl-transferase (GGT) and alkaline phosphatase (AP) significantly indicated the later therapy response (39 progression versus 32 no progression). In multivariate analysis, nucleosomes (24 h), AP (24 h) and TACE number were independent predictive markers. The risk score of this combination model achieved an AUC of 81.8% in receiver operating characteristic (ROC) curves and a sensitivity for prediction of non-response to therapy of 41% at 97% specificity, and of 72% at 78% specificity. CONCLUSION: Circulating nucleosomes and liver markers are valuable tools for early estimation of the efficacy of TACE therapy in HCC patients. PMID- 21615954 TI - Crystal structure of alkyl hydroperoxidase D like protein PA0269 from Pseudomonas aeruginosa: homology of the AhpD-like structural family. AB - BACKGROUND: Alkyl hydroperoxidase activity provides an important antioxidant defense for bacterial cells. The catalytic mechanism requires two peroxidases, AhpC and AhpD, where AhpD plays the role of an essential adaptor protein. RESULTS: The crystal structure of a putative AhpD from Pseudomonas aeruginosa has been determined at 1.9 A. The protein has an all-helical fold with a chain topology similar to a known AhpD from Mycobacterium tuberculosis despite a low overall sequence identity of 9%. A conserved two alpha-helical motif responsible for function is present in both. However, in the P. aeruginosa protein, helices H3, H4 of this motif are located at the N-terminal part of the chain, while in M. tuberculosis AhpD, the corresponding helices H8, H9 are situated at the C terminus. Residues 24-62 of the putative catalytic region of P. aeruginosa have a higher sequence identity of 33% where the functional activity is supplied by a proton relay system of five residues, Glu36, Cys48, Tyr50, Cys51, and His55, and one structural water molecule. A comparison of five other related hypothetical proteins from various species, assigned to the alkyl hydroperoxidase D-like protein family, shows they contain the same conserved structural motif and catalytic sequence Cys-X-X-Cys. We have shown that AhpD from P. aeruginosa exhibits a weak ability to reduce H(2)O(2) as tested using a ferrous oxidation xylenol orange (FOX) assay, and this activity is blocked by thiol alkylating reagents. CONCLUSION: Thus, this hypothetical protein was assigned to the AhpD like protein family with peroxidase-related activity. The functional relationship of specific oligomeric structures of AhpD-like structural family is discussed. PMID- 21615955 TI - A comparison of small monetary incentives to convert survey non-respondents: a randomized control trial. AB - BACKGROUND: Maximizing response rates is critically important in order to provide the most generalizable and unbiased research results. High response rates reduce the chance of respondents being systematically different from non-respondents, and thus, reduce the risk of results not truly reflecting the study population. Monetary incentives are often used to improve response rates, but little is known about whether larger incentives improve response rates in those who previously have been unenthusiastic about participating in research. In this study we compared the response rates and cost-effectiveness of a $5 versus $2 monetary incentive accompanying a short survey mailed to patients who did not respond or refused to participate in research study with a face-to-face survey. METHODS: 1,328 non-responders were randomly assigned to receive $5 or $2 and a short, 10 question survey by mail. Reminder postcards were sent to everyone; those not returning the survey were sent a second survey without incentive. Overall response rates, response rates by incentive condition, and odds of responding to the larger incentive were calculated. Total costs (materials, postage, and labor) and incremental cost-effectiveness ratios were also calculated and compared by incentive condition. RESULTS: After the first mailing, the response rate within the $5 group was significantly higher (57.8% vs. 47.7%, p<.001); after the second mailing, the difference narrowed by 80%, resulting in a non-significant difference in cumulative rates between the $5 and $2 groups (67.3% vs. 65.4%, respectively, p=.47). Regardless of incentive or number of contacts, respondents were significantly more likely to be male, white, married, and 50-75 years old. Total costs were higher with the larger versus smaller incentive ($13.77 versus $9.95 per completed survey). CONCLUSIONS: A $5 incentive provides a significantly higher response rate than a $2 incentive if only one survey mailing is used but not if two survey mailings are used. PMID- 21615956 TI - Impact of a probiotic fermented milk in the gut ecosystem and in the systemic immunity using a non-severe protein-energy-malnutrition model in mice. AB - BACKGROUND: Malnutrition affects the immune response, causing a decrease of defence mechanisms and making the host more susceptible to infections. Probiotics can reconstitute the intestinal mucosa and stimulate local and systemic immunity. The aim of this work was evaluate the effects of a probiotic fermented milk as a complement of a re-nutrition diet, on the recovery of the intestinal barrier, and mucosal and systemic immune functions in a murine model of non-severe protein energy-malnutrition. Its potential protection against Salmonella enterica serovar Typhimurium (S. Typhimurium) infection was also analyzed. METHODS: Mice were undernourished and divided into 3 groups according to the dietary supplement received during re-nutrition (milk, probiotic fermented milk or its bacterial free supernatant) and compared to well-nourished and malnourished mice. They were sacrificed previous to the re-nutrition and 5 days post re-nutrition. The phagocytic activity of macrophages from spleen and peritoneum and the changes in the intestinal histology and microbiota were evaluated. Different immune cell populations and cytokine productions were analyzed in the small intestine tissues. The effect of the re-nutrition supplements on the systemic immunity using OVA antigen and against an infection with S. Typhimurium was also studied. RESULTS: Probiotic fermented milk was the most effective re-nutrition diet that improved the intestinal microbiota. Its administration also increased the number of IgA+ cells, macrophages and dendritic cells. The production of different cytokine (IFN-gamma, TNF-alpha, IL-12) by these cells and the phagocytic activity in peritoneum and spleen was also increased. This re-nutrition diet also stimulated the systemic immune response against OVA antigen which was diminished after the malnutrition period and also improved the host response against S. Typhimurium, decreasing the spread of pathogenic bacteria to the liver and the spleen. The importance of the metabolites released during milk fermentation was also demonstrated through the analysis of the bacterial free supernatant obtained from the probiotic fermented milk, but the whole product showed the best effects in the parameters evaluated in this study. CONCLUSIONS: The administration of probiotic fermented milk as a dietary supplement during the re-nutrition process in a murine immunodeficiency model by malnutrition could be a good adjuvant diet to improve the gut and systemic immune response for the protection against Salmonella infection. PMID- 21615957 TI - Patterns and determinants of breastfeeding and complementary feeding practices in urban informal settlements, Nairobi Kenya. AB - BACKGROUND: The World Health Organisation (WHO) recommends exclusive breastfeeding during the first six months of life for optimal growth, development and health. Breastfeeding should continue up to two years or more and nutritionally adequate, safe, and appropriately-fed complementary foods should be introduced at the age of six months to meet the evolving needs of the growing infant. Little evidence exists on breastfeeding and infant feeding practices in urban slums in sub-Saharan Africa. Our aim was to assess breastfeeding and infant feeding practices in Nairobi slums with reference to WHO recommendations. METHODS: Data from a longitudinal study conducted in two Nairobi slums are used. The study used information on the first year of life of 4299 children born between September 2006 and January 2010. All women who gave birth during this period were interviewed on breastfeeding and complementary feeding practices at recruitment and this information was updated twice, at four-monthly intervals. Cox proportional hazard analysis was used to determine factors associated with cessation of breastfeeding in infancy and early introduction of complementary foods. RESULTS: There was universal breastfeeding with almost all children (99%) having ever been breastfed. However, more than a third (37%) were not breastfed in the first hour following delivery, and 40% were given something to drink other than the mothers' breast milk within 3 days after delivery. About 85% of infants were still breastfeeding by the end of the 11th month. Exclusive breastfeeding for the first six months was rare as only about 2% of infants were exclusively breastfed for six months. Factors associated with sub-optimal infant breastfeeding and feeding practices in these settings include child's sex; perceived size at birth; mother's marital status, ethnicity; education level; family planning (pregnancy desirability); health seeking behaviour (place of delivery) and; neighbourhood (slum of residence). CONCLUSIONS: The study indicates poor adherence to WHO recommendations for breastfeeding and infant feeding practices. Interventions and further research should pay attention to factors such as cultural practices, access to and utilization of health care facilities, child feeding education, and family planning. PMID- 21615958 TI - Angiopoietin-1 is associated with cerebral vasospasm and delayed cerebral ischemia in subarachnoid hemorrhage. AB - BACKGROUND: Angiopoietin-1 (Ang-1) and -2 (Ang-2) are keyplayers in the regulation of endothelial homeostasis and vascular proliferation. Angiopoietins may play an important role in the pathophysiology of cerebral vasospasm (CVS). Ang-1 and Ang-2 have not been investigated in this regard so far. METHODS: 20 patients with subarachnoid hemorrhage (SAH) and 20 healthy controls (HC) were included in this prospective study. Blood samples were collected from days 1 to 7 and every other day thereafter. Ang-1 and Ang-2 were measured in serum samples using commercially available enzyme-linked immunosorbent assay. Transcranial Doppler sonography was performed to monitor the occurrence of cerebral vasospasm. RESULTS: SAH patients showed a significant drop of Ang-1 levels on day 2 and 3 post SAH compared to baseline and HC. Patients, who developed Doppler sonographic CVS, showed significantly lower levels of Ang-1 with a sustained decrease in contrast to patients without Doppler sonographic CVS, whose Ang-1 levels recovered in the later course of the disease. In patients developing cerebral ischemia attributable to vasospasm significantly lower Ang-1 levels have already been observed on the day of admission. Differences of Ang-2 between SAH patients and HC or patients with and without Doppler sonographic CVS were not statistically significant. CONCLUSIONS: Ang-1, but not Ang-2, is significantly altered in patients suffering from SAH and especially in those experiencing CVS and cerebral ischemia. The loss of vascular integrity, regulated by Ang-1, might be in part responsible for the development of cerebral vasospasm and subsequent cerebral ischemia. PMID- 21615959 TI - An interesting journey of an ingested needle: a case report and review of the literature on extra-abdominal migration of ingested foreign bodies. AB - Swallowed foreign bodies encounter a major problem especially in children, but fortunately they mostly do not cause any related complication and are easily passed with the stool. In this paper, an interesting journey of a needle is presented. A 20-year old female admitted to our emergency service after she had swallowed a sewing machine needle, which is initially observed in the stomach in the plain abdominal radiography. During the follow-up period, the needle traveled through bowels, and surprisingly was observed in the left lung on 10th day of the follow-up. It was removed with a thoracotomy and pneumotomy under the fluoroscopic guidance. The postoperative period was uneventful and the patient was discharged from the hospital on the day 5. We also review the literature on interesting extra-abdominal migrations of swallowing foreign bodies. PMID- 21615960 TI - Antibacterial and dermal toxicological profiles of ethyl acetate extract from Crassocephalum bauchiense (Hutch.) Milne-Redh (Asteraceae). AB - BACKGROUND: The emergence in recent years of numerous resistant strains of pathogenic bacteria to a range of formerly efficient antibiotics constitutes a serious threat to public health. Crassocephalum bauchiense, a medicinal herb found in the West Region of Cameroon is used to treat gastrointestinal infections as well as liver disorders. The ethyl acetate extract from the leaves of C. bauchiense was evaluated for its antibacterial activity as well as acute and sub acute toxicities. METHODS: The plant extract was prepared by maceration in ethyl acetate. Its phytochemical screening was done by standard methods. The broth microdilution method was used to evaluate the in vitro antibacterial activity. The in vivo antibacterial activity of a gel formulation (0.05, 1 and 2% w/v) of this extract was evaluated using a Staphylococcus aureus-induced dermatitis in a murine model. Selected haematological and biochemical parameters were used to evaluate the dermal sub-acute toxicity of the extract in rats. RESULTS: Phytochemical screening of the C. bauchiense extract revealed the presence of alkaloids, phenols, tannins and sterols. In vitro antibacterial activities were observed against all the tested microorganisms (MIC = 0.04-6.25 mg/ml). Formulated extract-gel (2% w/v) and gentamycin (reference drug) eradicated the microbial infection after five days of treatment. A single dermal dose of this extract up to 32 g/kg body weight (bw) did not produce any visible sign of toxicity. Also, daily dermal application of the C. bauchiense extract gel formulation for 28 days did not show any negative effect, instead some biochemical parameters such as alanine aminotransferase (ALT and AST), low density lipoprotein (LDL), high density lipoprotein (HDL) and triglycerides were significantly (p < 0.05) affected positively. CONCLUSION: These results indicate that the C. bauchiense ethyl acetate extract can be used safely for the treatment of some bacterial infections. PMID- 21615961 TI - How do psychosocial determinants in migrant women in the Netherlands differ from these among their counterparts in their country of origin? A cross-sectional study. AB - BACKGROUND: Migration of non-Western women into Western countries often results in an increase in smoking prevalence among migrant women. To gain more insight into how to prevent this increase, we compared psychosocial determinants of smoking between Surinamese women in Suriname and those in the Netherlands. METHODS: Data were obtained between 2000 and 2004 from two cross-sectional studies, the CVRFO study in Suriname (n = 702) and the SUNSET study in the Netherlands (n = 674). For analyses of determinants, we collected additional data in CVRFO study population (n = 85). Differences between the two groups were analysed by chi-square analyses and logistic regression analyses. RESULTS: As was found in other studies among migrant women, more Surinamese migrant women in the Netherlands smoked (31%) than women in Suriname (16%). More Surinamese women in the Netherlands than in Suriname had a positive affective and cognitive attitude towards smoking (OR = 2.6 (95%CI 1.05;6.39) and OR = 3.3 (95%CI 1.31;8.41)). They perceived a positive norm within their partners and friends regarding smoking more frequently (OR = 6.5 (95%CI 2.7;15.6) and OR = 3.3 (95%CI 1.50;7.25)). CONCLUSION: Migrant women are more positive towards smoking and perceived a more positive norm towards smoking when compared with women in the country of origin. Interventions targeted at the psychosocial determinants regarding smoking for newly migrated women, in particular the consequences of smoking and the norm towards smoking might help to prevent an increase in smoking in those populations. PMID- 21615962 TI - Epstein-Barr virus myelitis and Castleman's disease in a patient with acquired immune deficiency syndrome: a case report. AB - INTRODUCTION: Few cases of Epstein-Barr virus myelitis have been described in the literature. Multi-centric Castleman's disease is a lymphoproliferative disorder that is well known for its associations with the human immunodeficiency virus, human herpes virus 8, and Kaposi's sarcoma. The concurrent presentation of these two diseases in a patient at the same time is extremely unusual. CASE PRESENTATION: We describe the case of a 43-year-old Caucasian man with acquired immune deficiency syndrome who presented with fever, weight loss and diffuse lymphadenopathy, and was diagnosed with multi-centric Castleman's disease. He presented three weeks later with lower extremity weakness and urinary retention, at which time cerebrospinal fluid contained lymphocytic pleocytosis and elevated protein. Magnetic resonance imaging demonstrated abnormal spinal cord signal intensity over several cervical and thoracic segments, suggesting the diagnosis of myelitis. Our patient was ultimately diagnosed with Epstein-Barr virus myelitis, as Epstein-Barr virus DNA was detected by polymerase chain reaction in the cerebrospinal fluid. CONCLUSION: To the best of our knowledge, this is the first case of multi-centric Castleman's disease followed by acute Epstein-Barr virus myelitis in a human immunodeficiency virus-infected patient. Clinicians caring for human immunodeficiency virus-infected patients should be vigilant about monitoring patients with increasing lymphadenopathy, prompting thorough diagnostic investigations when necessary. PMID- 21615963 TI - Methods used for successful follow-up in a large scale national cohort study in Thailand. AB - BACKGROUND: Ensuring successful follow-up is essential when conducting a prospective cohort study. Most existing literature reviewing methods to ensure a high response rate is based on experience in developed nations. FINDINGS: We report our 4-year follow-up success for a national cohort study examining the health transition underway in Thailand. We began the cohort study in 2005 with a baseline postal questionnaire sent to all 200,000 Thais enrolled as distance learning students at Sukhothai Thammathirat Open University and residing all over Thailand; 87,134 or 44% of the students responded. Subsequently we used University and national media to inform cohort members of study progress. Also, we prepared a health book with study results and health advice which was distributed to all cohort members. After 4 years we repeated the survey and achieved a 71% response rate. In this paper we report the methods used to achieve this responseThe initial follow-up mail-out generated a response rate of about 48% reflecting the extensive preparatory work between baseline and follow-up. After 4 rounds of telephone contact (more than 100,000 phone calls) and 4 related mail-out rounds progressively over 16 months an overall response rate was achieved of just over 71% (n = 60,774). The total cost was US$4.06/respondent - 19% for printing, 21% for postage, 14% for tape measures (included in mail-out), 18% for data processing 22% for prizes and 6% for telephone. CONCLUSIONS: Many of the methods reported as effective for mail questionnaire and cohort response rates held true for Thailand. These included being associated with a university, incentivating cooperation, follow-up contact, providing a second copy of questionnaire where necessary, and assurance of confidentiality. Telephone contact with the cohort and the small prizes given to responders were particularly important in the Thai context as was Thai leadership of the research team. PMID- 21615964 TI - Expression and localization of aquaporin 1b during oocyte development in the Japanese eel (Anguilla japonica). AB - To elucidate the molecular mechanisms underling hydration during oocyte maturation, we characterized the structure of Japanese eel (Anguilla japonica) novel-water selective aquaporin 1 (AQP1b) that thought to be involved in oocyte hydration. The aqp1b cDNA encodes a 263 amino acid protein that includes the six potential transmembrane domains and two Asn-Pro-Ala motifs. Reverse transcription polymerase chain reaction showed transcription of Japanese eel aqp1b in ovary and testis but not in the other tissues. In situ hybridization studies with the eel aqp1b cRNA probe revealed intense eel aqp1b signal in the oocytes at the perinucleolus stage and the signals became faint during the process of oocyte development. Light microscopic immunocytochemical analysis of ovary revealed that the Japanese eel AQP1b was expressed in the cytoplasm around the yolk globules which were located in the peripheral region of oocytes during the primary yolk globule stage; thereafter, the immunoreactivity was observed throughout the cytoplasm of oocyte as vitellogenesis progressed. The immunoreactivity became localized around the large membrane-limited yolk masses which were formed by the fusion of yolk globules during the oocyte maturation phase. These results together indicate that AQP1b, which is synthesized in the oocyte during the process of oocyte growth, is essential for mediating water uptake into eel oocytes. PMID- 21615965 TI - P53 in human melanoma fails to regulate target genes associated with apoptosis and the cell cycle and may contribute to proliferation. AB - BACKGROUND: Metastatic melanoma represents a major clinical problem. Its incidence continues to rise in western countries and there are currently no curative treatments. While mutation of the P53 tumour suppressor gene is a common feature of many types of cancer, mutational inactivation of P53 in melanoma is uncommon; however, its function often appears abnormal. METHODS: In this study whole genome bead arrays were used to examine the transcript expression of P53 target genes in extracts from 82 melanoma metastases and 6 melanoma cell lines, to provide a global assessment of aberrant P53 function. The expression of these genes was also examined in extracts derived from diploid human melanocytes and fibroblasts. RESULTS: The results indicated that P53 target transcripts involved in apoptosis were under-expressed in melanoma metastases and melanoma cell lines, while those involved in the cell cycle were over-expressed in melanoma cell lines. There was little difference in the transcript expression of P53 target genes between cell lines with null/mutant P53 compared to those with wild-type P53, suggesting that altered expression in melanoma was not related to P53 status. Similarly, down-regulation of P53 by short-hairpin RNA (shRNA) had limited effect on P53 target gene expression in melanoma cells, whereas there were a large number of P53 target genes whose mRNA expression was significantly altered by P53 inhibition in melanocytes. Analysis of whole genome gene expression profiles indicated that the ability of P53 to regulate genes involved in the cell cycle was significantly reduced in melanoma cells. Moreover, inhibition of P53 in melanocytes induced changes in gene expression profiles that were characteristic of melanoma cells and resulted in increased proliferation. Conversely, knockdown of P53 in melanoma cells resulted in decreased proliferation. CONCLUSIONS: These results indicate that P53 target genes involved in apoptosis and cell cycle regulation are aberrantly expressed in melanoma and that this aberrant functional activity of P53 may contribute to the proliferation of melanoma. PMID- 21615966 TI - The evolution of reproductive isolation in a simultaneous hermaphrodite, the freshwater snail Physa. AB - BACKGROUND: The cosmopolitan freshwater snail Physa acuta has recently found widespread use as a model organism for the study of mating systems and reproductive allocation. Mitochondrial DNA phylogenies suggest that Physa carolinae, recently described from the American southeast, is a sister species of P. acuta. The divergence of the acuta/carolinae ancestor from the more widespread P. pomilia appears to be somewhat older, and the split between a hypothetical acuta/carolinae/pomilia ancestor and P. gyrina appears older still. RESULTS: Here we report the results of no-choice mating experiments yielding no evidence of hybridization between gyrina and any of four other populations (pomilia, carolinae, Philadelphia acuta, or Charleston acuta), nor between pomilia and carolinae. Crosses between pomilia and both acuta populations yielded sterile F1 progeny with reduced viability, while crosses between carolinae and both acuta populations yielded sterile F1 hybrids of normal viability. A set of mate-choice tests also revealed significant sexual isolation between gyrina and all four of our other Physa populations, between pomilia and carolinae, and between pomilia and Charleston acuta, but not between pomilia and the acuta population from Philadelphia, nor between carolinae and either acuta population. These observations are consistent with the origin of hybrid sterility prior to hybrid inviability, and a hypothesis that speciation between pomilia and acuta may have been reinforced by selection for prezygotic reproductive isolation in sympatry. CONCLUSIONS: We propose a two-factor model for the evolution of postzygotic reproductive incompatibility in this set of five Physa populations consistent with the Dobzhansky-Muller model of speciation, and a second two-factor model for the evolution of sexual incompatibility. Under these models, species trees may be said to correspond with gene trees in American populations of the freshwater snail, Physa. PMID- 21615967 TI - Systems mapping: how to improve the genetic mapping of complex traits through design principles of biological systems. AB - BACKGROUND: Every phenotypic trait can be viewed as a "system" in which a group of interconnected components function synergistically to yield a unified whole. Once a system's components and their interactions have been delineated according to biological principles, we can manipulate and engineer functionally relevant components to produce a desirable system phenotype. RESULTS: We describe a conceptual framework for mapping quantitative trait loci (QTLs) that control complex traits by treating trait formation as a dynamic system. This framework, called systems mapping, incorporates a system of differential equations that quantifies how alterations of different components lead to the global change of trait development and function through genes, and provides a quantitative and testable platform for assessing the interplay between gene action and development. We applied systems mapping to analyze biomass growth data in a mapping population of soybeans and identified specific loci that are responsible for the dynamics of biomass partitioning to leaves, stem, and roots. CONCLUSIONS: We show that systems mapping implemented by design principles of biological systems is quite versatile for deciphering the genetic machineries for size shape, structural-functional, sink-source and pleiotropic relationships underlying plant physiology and development. Systems mapping should enable geneticists to shed light on the genetic complexity of any biological system in plants and other organisms and predict its physiological and pathological states. PMID- 21615968 TI - Treating postnatal depressive symptoms in primary care: a randomised controlled trial of GP management, with and without adjunctive counselling. AB - BACKGROUND: Postnatal depression (PND) is under-diagnosed and most women do not access effective help. We aimed to evaluate comparative management of (PND) following screening with the Edinburgh Postnatal Depression Scale, using three best-practice care pathways by comparing management by general practitioners (GPs) alone compared to adjunctive counselling, based on cognitive behavioural therapy (CBT), delivered by postnatal nurses or psychologists. METHODS: This was a parallel, three-group randomised controlled trial conducted in a primary care setting (general practices and maternal & child health centres) and a psychology clinic. A total of 3,531 postnatal women were screened for symptoms of depression; 333 scored above cut-off on the screening tool and 169 were referred to the study. Sixty-eight of these women were randomised between the three treatment groups. RESULTS: Mean scores on the Beck Depression Inventory (BDI-II) at entry were in the moderate-to-severe range. There was significant variation in the post-study frequency of scores exceeding the threshold indicative of mild-to severe depressive symptoms, such that more women receiving only GP management remained above the cut-off score after treatment (p = .028). However, all three treatment conditions were accompanied by significant reductions in depressive symptoms and mean post-study BDI-II scores were similar between groups. Compliance was high in all three groups. Women rated the treatments as highly effective. Rates of both referral to the study (51%), and subsequent treatment uptake (40%) were low. CONCLUSIONS: Data from this small study suggest that GP management of PND when augmented by a CBT-counselling package may be successful in reducing depressive symptoms in more patients compared to GP management alone. The relatively low rates of referral and treatment uptake, suggest that help seeking remains an issue for many women with PND, consistent with previous research. PMID- 21615969 TI - Quality of life at the Dead Sea region: the lower the better? An observational study. AB - BACKGROUND: The Dead Sea region, the lowest in the world at 410 meters below sea level, is considered a potent climatotherapy center for the treatment of different chronic diseases. OBJECTIVE: To assess the prevalence of chronic diseases and the quality of life of residents of the Dead Sea region compared with residents of the Ramat Negev region, which has a similar climate, but is situated 600 meters above sea level. METHODS: An observational study based on a self-administered questionnaire. Data were collected from kibbutz (communal settlement) members in both regions. Residents of the Dead Sea were the study group and of Ramat Negev were the control group. We compared demographic characteristics, the prevalence of different chronic diseases and health-related quality of life (HRQOL) using the SF-36 questionnaire. RESULTS: There was a higher prevalence of skin nevi and non-inflammatory rheumatic diseases (NIRD) among Dead Sea residents, but they had significantly higher HRQOL mean scores in general health (68.7 +/- 21 vs. 64.4 +/- 22, p = 0.023) and vitality (64.7 +/- 17.9 vs. 59.6 +/- 17.3, p = 0.001), as well as significantly higher summary scores: physical component score (80.7 +/- 18.2 vs. 78 +/- 18.6, p = 0.042), and mental component score (79 +/- 16.4 vs. 77.2 +/- 15, p = 0.02). These results did not change after adjusting for social-demographic characteristics, health-related habits, and chronic diseases. CONCLUSIONS: No significant difference between the groups was found in the prevalence of most chronic diseases, except for higher rates of skin nevi and NIRD among Dead Sea residents. HRQOL was significantly higher among Dead Sea residents, both healthy or with chronic disease. PMID- 21615970 TI - Adhesive polypeptides of Staphylococcus aureus identified using a novel secretion library technique in Escherichia coli. AB - BACKGROUND: Bacterial adhesive proteins, called adhesins, are frequently the decisive factor in initiation of a bacterial infection. Characterization of such molecules is crucial for the understanding of bacterial pathogenesis, design of vaccines and development of antibacterial drugs. Because adhesins are frequently difficult to express, their characterization has often been hampered. Alternative expression methods developed for the analysis of adhesins, e.g. surface display techniques, suffer from various drawbacks and reports on high-level extracellular secretion of heterologous proteins in Gram-negative bacteria are scarce. These expression techniques are currently a field of active research. The purpose of the current study was to construct a convenient, new technique for identification of unknown bacterial adhesive polypeptides directly from the growth medium of the Escherichia coli host and to identify novel proteinaceous adhesins of the model organism Staphylococcus aureus. RESULTS: Randomly fragmented chromosomal DNA of S. aureus was cloned into a unique restriction site of our expression vector, which facilitates secretion of foreign FLAG-tagged polypeptides into the growth medium of E. coli DeltafliCDeltafliD, to generate a library of 1663 clones expressing FLAG-tagged polypeptides. Sequence and bioinformatics analyses showed that in our example, the library covered approximately 32% of the S. aureus proteome. Polypeptides from the growth medium of the library clones were screened for binding to a selection of S. aureus target molecules and adhesive fragments of known staphylococcal adhesins (e.g coagulase and fibronectin-binding protein A) as well as polypeptides of novel function (e.g. a universal stress protein and phosphoribosylamino-imidazole carboxylase ATPase subunit) were detected. The results were further validated using purified His-tagged recombinant proteins of the corresponding fragments in enzyme-linked immunoassay and surface plasmon resonance analysis. CONCLUSIONS: A new technique for identification of unknown bacterial adhesive polypeptides was constructed. Application of the method on S. aureus allowed us to identify three known adhesins and in addition, five new polypeptides binding to human plasma and extracellular matrix proteins. The method, here used on S. aureus, is convenient due to the use of soluble proteins from the growth medium and can in principle be applied to any bacterial species of interest. PMID- 21615972 TI - MixtureTree: a program for constructing phylogeny. AB - BACKGROUND: MixtureTree v1.0 is a Linux based program (written in C++) which implements an algorithm based on mixture models for reconstructing phylogeny from binary sequence data, such as single-nucleotide polymorphisms (SNPs). In addition to the mixture algorithm with three different optimization options, the program also implements a bootstrap procedure with majority-rule consensus. RESULTS: The MixtureTree program written in C++ is a Linux based package. The User's Guide and source codes will be available at http://math.asu.edu/~scchen/MixtureTree.html CONCLUSIONS: The efficiency of the mixture algorithm is relatively higher than some classical methods, such as Neighbor-Joining method, Maximum Parsimony method and Maximum Likelihood method. The shortcoming of the mixture tree algorithms, for example timing consuming, can be improved by implementing other revised Expectation-Maximization(EM) algorithms instead of the traditional EM algorithm. PMID- 21615973 TI - Australasian podiatry council conference 2011 melbourne, australia. 26-29 april 2011. Abstracts. PMID- 21615971 TI - Exacerbation of cigarette smoke-induced pulmonary inflammation by Staphylococcus aureus enterotoxin B in mice. AB - BACKGROUND: Cigarette smoke (CS) is a major risk factor for the development of COPD. CS exposure is associated with an increased risk of bacterial colonization and respiratory tract infection, because of suppressed antibacterial activities of the immune system and delayed clearance of microbial agents from the lungs. Colonization with Staphylococcus aureus results in release of virulent enterotoxins, with superantigen activity which causes T cell activation. OBJECTIVE: To study the effect of Staphylococcus aureus enterotoxin B (SEB) on CS induced inflammation, in a mouse model of COPD. METHODS: C57/Bl6 mice were exposed to CS or air for 4 weeks (5 cigarettes/exposure, 4x/day, 5 days/week). Endonasal SEB (10 MUg/ml) or saline was concomitantly applied starting from week 3, on alternate days. 24 h after the last CS and SEB exposure, mice were sacrificed and bronchoalveolar lavage (BAL) fluid and lung tissue were collected. RESULTS: Combined exposure to CS and SEB resulted in a raised number of lymphocytes and neutrophils in BAL, as well as increased numbers of CD8+ T lymphocytes and granulocytes in lung tissue, compared to sole CS or SEB exposure. Moreover, concomitant CS/SEB exposure induced both IL-13 mRNA expression in lungs and goblet cell hyperplasia in the airway wall. In addition, combined CS/SEB exposure stimulated the formation of dense, organized aggregates of B- and T- lymphocytes in lungs, as well as significant higher CXCL-13 (protein, mRNA) and CCL19 (mRNA) levels in lungs. CONCLUSIONS: Combined CS and SEB exposure aggravates CS-induced inflammation in mice, suggesting that Staphylococcus aureus could influence the pathogenesis of COPD. PMID- 21615975 TI - The role of wildlife in transboundary animal diseases. AB - This paper identifies some of the more important diseases at the wildlife livestock interface and the role wildlife plays in disease transmission. Domestic livestock, wildlife and humans share many similar pathogens. Pathogens of wild or domestic animal origin that can cause infections in humans are known as zoonotic organisms and the converse are termed as anthroponotic organisms. Seventy-seven percent of livestock pathogens and 91% of domestic carnivore pathogens are known to infect multiple hosts, including wildlife. Understanding this group of pathogens is critical to public health safety, because they infect a wide range of hosts and are most likely to emerge as novel causes of infection in humans and domestic animals. Diseases at the wildlife-livestock interface, particularly those that are zoonotic, must be an area of focus for public health programs and surveillance for emerging infectious diseases. Additionally, understanding wildlife and their role is a vital part of understanding the epidemiology and ecology of diseases. To do this, a multi-faceted approach combining capacity building and training, wildlife disease surveillance, wildlife-livestock interface and disease ecology studies, data and information sharing and outbreak investigation are needed. PMID- 21615974 TI - Chronic fatigue syndrome. AB - INTRODUCTION: Chronic fatigue syndrome (CFS) affects between 0.006% and 3% of the population depending on the criteria of definition used, with women being at higher risk than men. METHODS AND OUTCOMES: We conducted a systematic review and aimed to answer the following clinical question: What are the effects of treatments for chronic fatigue syndrome? We searched: Medline, Embase, The Cochrane Library, and other important databases up to March 2010 (Clinical Evidence reviews are updated periodically; please check our website for the most up-to-date version of this review). We included harms alerts from relevant organisations such as the US Food and Drug Administration (FDA) and the UK Medicines and Healthcare products Regulatory Agency (MHRA). RESULTS: We found 46 systematic reviews, RCTs, or observational studies that met our inclusion criteria. We performed a GRADE evaluation of the quality of evidence for interventions. CONCLUSIONS: In this systematic review we present information relating to the effectiveness and safety of the following interventions: antidepressants, cognitive behavioural therapy (CBT), corticosteroids, dietary supplements, evening primrose oil, galantamine, graded exercise therapy, homeopathy, immunotherapy, intramuscular magnesium, oral nicotinamide adenine dinucleotide, and prolonged rest. PMID- 21615976 TI - Fontan conversion with concomitant arrhythmia surgery for the failing atriopulmonary connections: mid-term results from a single centre. AB - OBJECTIVES: Classical Atriopulmonary Fontan connections tend to fail in the long term due to progressive anastomotic site obstruction, right atrial enlargement, and refractory atrial arrhythmias. Conversion to total cavopulmonary connection with concomitant arrhythmia surgery is a promising treatment but optimal timing of the procedure remains controversial. METHODS: Between the years 2002 and 2009, 15 patients with a median age of 26.2 (12-43) years underwent Fontan conversion operation with concomitant arrhythmia surgery. All were symptomatic and 14 out of the 15 patients had refractory arrhythmias. The duration of pre-operative arrhythmia and the outcome of surgery were correlated to study the impact of delay in surgical intervention on post-operative survival and arrhythmia control. RESULTS: There were two patients who died in the early post-operative period (13.3%). At the mid-term follow-up, 53 (20-86) months, late atrial arrhythmias had recurred in two of the 13 surviving patients (15.30%) and one patient developed late sinus node dysfunction. The need for anti-arrhythmic drugs decreased considerably from 93.5% to 15.3% on mid-term follow-up. There was no late death or need for cardiac transplantation. The duration of arrhythmia before surgery was prolonged for more than 10 years in patients who died as well as in those who had complications like late recurrence of arrhythmias, dependence on anti-arrhythmic medications, and worsening of ventricular dysfunction. CONCLUSIONS: Fontan conversion is a well-established treatment option for salvaging the failing atriopulmonary connections. Concomitant arrhythmia surgery effectively resolves the refractory atrial arrhythmias and improves survival, but we need to optimise the timing of Fontan conversion to improve the long-term outcome. PMID- 21615977 TI - Agraphia in Korean patients with early onset Alzheimer's disease. AB - BACKGROUND: Agraphia in Korean patients may be different from agraphia in other patients who use alphabetical writing systems due to the "visuoconstructional script" characteristics of the Korean writing system, Hangul. Patients with early onset Alzheimer's disease (EOAD) have a severe degree of hypometabolism in the parietal area, which is known to be involved in processing visuospatial function. Thus, we explored the diverse error patterns manifested in writing single syllables in Korean patients with EOAD. METHODS: A study sample of 35 patients with EOAD and 18 healthy controls (HC) performed a Hangul writing task. We analyzed the erroneous responses of the subjects according to visuoconstructional and linguistic characteristics. In addition, we evaluated the relationship between Hangul writing and the neuropsychological variables as well as the severity of dementia. RESULTS: When comparing the total number of erroneous responses between EOAD and HC groups, the performances of EOAD patients were significantly worse than those of HC. EOAD patients demonstrated visuoconstructional errors even in the early stages of the disease. Severity of dementia and multiple cognitive domains such as attention, language, immediate memory, and frontal executive functions significantly correlated with the performance of Hangul writing. CONCLUSION: Our findings suggest that patients with EOAD exhibit not only linguistic errors but also visuoconstructional manifestations of agraphia, which are associated with cognitive impairments in the multiple domains. PMID- 21615978 TI - The external ion microbeam of the LABEC laboratory in florence: some applications to cultural heritage. AB - Ion beam analysis (IBA) techniques are a powerful analytical tool used to investigate the composition and structure of precious materials principally because they can be applied in atmosphere. Thus, the sample can be analyzed as is, and heating and charging effects are strongly diminished. Since IBA measurements can be made with low ion currents and acquisition time, the damage risk is limited. At the microbeam line of the LABEC laboratory, it is possible to exploit the potentials of IBA techniques in an external set up to reconstruct the distribution maps of all the detected elements over the analyzed area with spatial resolutions as low as 10 MUm. This is an important feature when objects with inhomogeneous structures-on a scale of hundred microns or so-are investigated, as happens in some cases with artworks. The detection set up installed on our external microbeam allows us to use different IBA techniques simultaneously. Thus, in a single measurement run, it is possible to obtain complementary information on both sample composition and structure. Some applications to works of art are presented here as examples of the analytical capabilities of the external scanning microbeam in the cultural heritage field. PMID- 21615979 TI - Compositional analysis with atomic column spatial resolution by 5th-order aberration-corrected scanning transmission electron microscopy. AB - We show in this article that it is possible to obtain elemental compositional maps and profiles with atomic-column resolution across an InxGa1-xAs multilayer structure from 5th-order aberration-corrected high-angle annular dark-field scanning transmission electron microscopy (HAADF-STEM) images. The compositional profiles obtained from the analysis of HAADF-STEM images describe accurately the distribution of In in the studied multilayer in good agreement with Muraki's segregation model [Muraki, K., Fukatsu, S., Shiraki, Y. & Ito, R. (1992). Surface segregation of In atoms during molecular beam epitaxy and its influence on the energy levels in InGaAs/GaAs quantums wells. Appl Phys Lett 61, 557-559]. PMID- 21615980 TI - Carbon diffusion from methane into walls of carbon nanotube through structurally and compositionally modified iron catalyst. AB - To understand diffusion processes occurring inside Fe catalysts during multiwall carbon nanotube (MWCNT) growth, catalysts were studied using atomic-resolution scanning transmission electron microscopy combined with electron energy-loss spectroscopy. Nanotube walls emanate from structurally modified and chemically complex catalysts that consist of cementite and a 5 nm amorphous FeOx cap separated by a 2-3 nm thick carbon-rich region that also contains Fe and O (a C:FexOy). Nonuniform distribution of carbon atoms throughout the catalyst base reveals that carbon molecules from the gas phase decompose near the catalyst multisection junction, where the MWCNT walls terminate. Formation of the a C:FexOy region provides the essential carbon source for MWCNT growth. Two different carbon diffusion mechanisms are responsible for the growth of the inner and outer walls of each MWCNT. PMID- 21615981 TI - Portable apparatus for in situ x-ray diffraction and fluorescence analyses of artworks. AB - A portable X-ray fluorescence/X-ray diffraction (XRF/XRD) system for artwork studies has been designed constructed and tested. It is based on Debye Scherrer XRD in reflection that takes advantage of many recent improvements in the handling of X-rays (polycapillary optics; advanced two-dimensional detection). The apparatus is based on a copper anode air cooled X-ray source, and the XRD analysis is performed on a 5-20 MUm thick layer from the object surface. Energy dispersive XRF elemental analysis can be performed at the same point as XRD, giving elemental compositions that support the interpretation of XRD diagrams. XRF and XRD analyses were tested to explore the quality and the limits of the analytical technique. The XRD diagrams are comparable in quality with diagrams obtained with conventional laboratory equipment. The mineral identification of materials in artwork is routinely performed with the portable XRF-XRD system. Examples are given for ceramic glazes containing crystals and for paintings where the determination of pigments is still a challenge for nondestructive analysis. For instance, lead compounds that provide a variety of color pigments can be easily identified as well as a pigment such as lapis lazuli that is difficult to identify by XRF alone. More than 70 works of art have been studied in situ in museums, monuments, etc. In addition to ceramics and paintings, these works include bronzes, manuscripts, etc., which permit improvement in the comprehension of ancient artistic techniques. PMID- 21615982 TI - Genomic approaches for the understanding of aging in model organisms. AB - Aging is one of the most complicated biological processes in all species. A number of different model organisms from yeast to monkeys have been studied to understand the aging process. Until recently, many different age-related genes and age-regulating cellular pathways, such as insulin/IGF-1-like signal, mitochondrial dysfunction, Sir2 pathway, have been identified through classical genetic studies. Parallel to genetic approaches, genome-wide approaches have provided valuable insights for the understanding of molecular mechanisms occurring during aging. Gene expression profiling analysis can measure the transcriptional alteration of multiple genes in a genome simultaneously and is widely used to elucidate the mechanisms of complex biological pathways. Here, current global gene expression profiling studies on normal aging and age-related genetic/environmental interventions in widely-used model organisms are briefly reviewed. PMID- 21615983 TI - Mitochondrial superoxide anion (O(2)(-)) inducible "mev-1" animal models for aging research. AB - Most intracellular reactive oxygen species (ROS), especially superoxide anion (O(2)(-)) that is converted from oxygen, are overproduced by excessive electron leakage from the mitochondrial respiratory chain. Intracellular oxidative stress that damages cellular components can contribute to lifestyle-related diseases such as diabetes and arteriosclerosis, and age-related diseases such as cancer and neuronal degenerative diseases. We have previously demonstrated that the excessive mitochondrial O(2)(-) production caused by SDHC mutations (G71E in C. elegans, I71E in Drosophila and V69E in mouse) results in premature death in C. elegans and Drosophila, cancer in mouse embryonic fibroblast cells and infertility in transgenic mice. SDHC is a subunit of mitochondrial complex II. In humans, it has been reported that mutations in SDHB, SDHC or SDHD often result in inherited head and neck paragangliomas (PGLs). Recently, we established Tet-mev-1 conditional transgenic mice using our uniquely developed Tet-On/Off system, which equilibrates transgene expression to endogenous levels. These mice experienced mitochondrial respiratory chain dysfunction that resulted in O(2)(-) overproduction. The mitochondrial oxidative stress caused excessive apoptosis leading to low birth weight and growth retardation in the neonatal developmental phase in Tet-mev-1 mice. Here, we briefly describe the relationships between mitochondrial O(2)(-) and aging phenomena in mev-1 animal models. [BMB reports 2011; 44(5): 298-305]. PMID- 21615984 TI - Alteration in NCX-3 immunoreactivity within the gerbil hippocampus following spontaneous seizures. AB - Although NCX-3 is highly expressed in the brain, the distribution of NCX-3 in the epileptic hippocampus is still controversial. Therefore, to assess the distribution and pattern of NCX-3 expression in epileptic hippocampus, we performed a comparative analysis of NCX-3 immunoreactivities in the hippocampus of seizure-resistant (SR) and seizure-sensitive (SS) gerbils. In SR gerbils, NCX 3 immunoreactivity was higher than pre-seizure SS gerbils, particularly in the pavalbumin (PV)-positive interneurons. Three h post-ictal, NCX-3 immunoreactivity in the SS gerbil hippocampus was markedly elevated to the level of SR gerbils. Six h post-ictal, the expression of NCX-3 was reduced to the level of pre-seizure SS gerbils. Therefore, the results of the present study suggest that down regulation of NCX-3 expression in the SS gerbil hippocampus may be involved in the hyperexcitability of SS gerbils due to an imbalance of intracellular Na(+)/Ca(2+) homeostasis and Ca(2+) concentration. PMID- 21615985 TI - Knockdown of cytosolic NADP(+) -dependent isocitrate dehydrogenase enhances MPP(+) -induced oxidative injury in PC12 cells. AB - 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and its toxic metabolite 1 methyl-4-phenylpyridium ion (MPP(+)) have been shown to induce Parkinson's disease-like symptoms as well as neurotoxicity in humans and animal species. Recently, we reported that maintenance of redox balance and cellular defense against oxidative damage are primary functions of the novel antioxidant enzyme cytosolic NADP(+) -dependent isocitrate dehydrogenase (IDPc). In this study, we examined the role of IDPc in cellular defense against MPP(+) -induced oxidative injury using PC12 cells transfected with IDPc small interfering RNA (siRNA). Our results demonstrate that MPP(+) -mediated disruption of cellular redox status, oxidative damage to cells, and apoptotic cell death were significantly enhanced by knockdown of IDPc. PMID- 21615986 TI - Distinct mutations in MLH1 and MSH2 genes in hereditary non-polyposis colorectal cancer (HNPCC) families from China. AB - Hereditary non-polyposis Colorectal Cancer (HNPCC) is an autosomal dominant inheritance syndrome. HNPCC is the most common hereditary variant of colorectal cancer (CRC), which accounts for 2-5% CRCs, mainly due to hMLH1 and hMSH2 mutations that impair DNA repair functions. Our study aimed to identify the patterns of hMSH2 and hMLH1 mutations in Chinese HNPCC patients. Ninety-eight unrelated families from China meeting Amsterdam or Bethesda criteria were included in our study. Germline mutations in MLH1 and MSH2 genes, located in the exons and the splice-site junctions, were screened in the 98 probands by direct sequencing. Eleven mutations were found in ten patients (11%), with six in MLH1 (54.5%) and five in MSH2 (45.5%) genes. One patient had mutations in both MLH1 and MSH2 genes. Three novel mutations in MLH1 gene (c.157_160delGAGG, c.2157dupT and c.-64G>T) were found for the first time, and one suspected hotspot in MSH2 (c.1168C>T) was revealed. PMID- 21615987 TI - Two groups of S-layer proteins, SLP1s and SLP2s, in Bacillus thuringiensis co exist in the S-layer and in parasporal inclusions. AB - We screened four B. thuringiensis strains whose parasporal inclusions contained the S-layer protein (SLP), and cloned two slp genes from each strain. Phylogenetic analysis indicated these SLPs could be divided into two groups, SLP1s and SLP2s. To confirm whether SLPs were present in the S-layer or as a parasporal inclusion, strains CTC and BMB1152 were chosen for further study. Western blots with whole-cell associated proteins from strains CTC and BMB1152 in the vegetative phase showed that SLP1s and SLP2s were constituents of the S layer. Immunofluorescence utilizing spore-inclusion mixtures of strains CTC and BMB1152 in the sporulation phase showed that SLP1s and SLP2s were also constituents of parasporal inclusions. When heterogeneously expressed in the crystal negative strain BMB171, four SLPs from strains CTC and BMB1152 could also form parasporal inclusions. This temporal and spatial expression is not an occasional phenomenon but ubiquitous in B. thuringiensis strains. PMID- 21615988 TI - Levosulpiride, (S)-(-)-5-Aminosulfonyl-N-[(1-ethyl-2-pyrrolidinyl) methyl]-2 methoxybenzamide, enhances the transduction efficiency of PEP-1-ribosomal protein S3 in vitro and in vivo. AB - Many proteins with poor transduction efficiency were reported to be delivered to cells by fusion with protein transduction domains (PTDs). In this study, we investigated the effect of levosulpiride on the transduction of PEP-1 ribosomal protein S3 (PEP-1-rpS3), and examined its influence on the stimulation of the therapeutic properties of PEP-1-rpS3. PEP-1-rpS3 transduction into HaCaT human keratinocytes and mouse skin was stimulated by levosulpiride in a manner that did not directly affect the cell viability. Following 12-O-tetradecanoylphorbol- 13 acetate (TPA)-induced inflammation in mice, levosulpiride alone was ineffective in reducing TPA-induced edema and in inhibiting the elevated productions of inflammatory mediators and cytokines, such as cyclooxygenase-2, inducible nitric oxide synthase, interleukin-6 and -1beta, and tumor necrosis factor- alpha. Anti inflammatory activity by PEP-1-rpS3 + levosulpiride was significantly more potent than by PEP-1-rpS3 alone. These results suggest that levosulpiride may be useful for enhancing the therapeutic effect of PEP-1-rpS3 against various inflammatory diseases. [BMB reports 2011; 44(5): 329-334]. PMID- 21615989 TI - Adenosine derived from Staphylococcus aureus-engulfed macrophages functions as a potent stimulant for the induction of inflammatory cytokines in mast cells. AB - In this study, we attempted to isolate novel mast cell-stimulating molecules from Staphylococcus aureus. Water-soluble extract of S. aureus cell lysate strongly induced human interleukin- 8 in human mast cell line-1 and mouse interleukin-6 in mouse bone marrow-derived mast cells. The active molecule was purified to homogeneity through a C(18) reverse phase HPLC column. By determination of its structure by MALDITOF and (1)H- and (13)C-NMR, adenosine was revealed to be responsible for the observed cytokine induction activities. Further studies using 8-sulfophenyl theophylline, a selective adenosine receptor blocker, verified that purified adenosine can induce interleukin-8 production via adenosine receptors on mast cells. Moreover, adenosine was purified from S. aureusengulfed RAW264.7 cells, a murine macrophage cell line, used to induce phagocytosis of S. aureus. These results show a novel view of the source of exogenous adenosine in vivo and provide a mechanistic link between inflammatory disease and bacterial infection. PMID- 21615990 TI - The stimulatory effect of CaCl(2), NaCl and NH(4)NO(3) salts on the ssDNA-binding activity of RecA depends on nucleotide cofactor and buffer pH. AB - The single-stranded DNA binding activity of the Escherichia coli RecA protein is crucial for homologous recombination to occur. This and other biochemical activities of ssDNA binding proteins may be affected by various factors. In this study, we analyzed the effect of CaCl(2), NaCl and NH(4)NO(3) salts in combination with the pH and nucleotide cofactor effect on the ssDNA-binding activity of RecA. The studies revealed that, in addition to the inhibitory effect, these salts exert also a stimulatory effect on RecA. These effects occur only under very strict conditions, and the presence or absence and the type of nucleotide cofactor play here a major role. It was observed that in contrast to ATP, ATPgammaS prevented the inhibitory effect of NaCl and NH(4)NO(3), even at very high salt concentration. These results indicate that ATPgammaS most likely stabilizes the structure of RecA required for DNA binding, making it resistant to high salt concentrations. PMID- 21615991 TI - An integrated bioinformatics analysis of mouse testis protein profiles with new understanding. AB - The testis is major male gonad responsible for spermatogenesis and steroidogenesis. Much knowledge is still remained to be learned about the control of these events. In this study, we performed a comprehensive bioinformatics analysis on 1,196 mouse testis proteins screened from public protein database. Integrated function and pathway analysis were performed through Database for Annotation, Visualization and Integrated Discovery (DAVID) and ingenuity Pathway Analysis (IPA), and significant features were clustered. Protein membrane organization and gene density on chromosomes were analyzed and discussed. The enriched bioinformatics analysis could provide clues and basis to the development of diagnostic markers and therapeutic targets for infertility and male contraception. PMID- 21615992 TI - Knock-down of human MutY homolog (hMYH) decreases phosphorylation of checkpoint kinase 1 (Chk1) induced by hydroxyurea and UV treatment. AB - The effect of human MutY homolog (hMYH) on the activation of checkpoint proteins in response to hydroxyurea (HU) and ultraviolet (UV) treatment was investigated in hMYH-disrupted HEK293 cells. hMYH-disrupted cells decreased the phosphorylation of Chk1 upon HU or UV treatment and increased the phosphorylation of Cdk2 and the amount of Cdc25A, but not Cdc25C. In siMYH-transfected cells, the increased rate of phosphorylated Chk1 upon HU or UV treatment was lower than that in siGFP-transfected cells, meaning that hMYH was involved in the activation mechanism of Chk1 upon DNA damage. The phosphorylation of ataxia telangiectasia and Rad3- related protein (ATR) upon HU or UV treatment was decreased in hMYH disrupted HEK293 and HaCaT cells. Co-immunoprecipitation experiments showed that hMYH was immunoprecipitated by anti-ATR. These results suggest that hMYH may interact with ATR and function as a mediator of Chk1 phosphorylation in response to DNA damage. PMID- 21615993 TI - [Histologic evaluation of malignant lymphoma]. PMID- 21615994 TI - [Primary gastrointestinal diffuse large B-cell lymphoma: an immunohistochemical and prognostic study of 90 cases]. AB - OBJECTIVE: To study the immunophenotype and prognostic significance of primary gastrointestinal diffuse large B-cell lymphoma, with reference to Hans, Choi and Tally algorithms. METHODS: The clinicopathologic features and follow-up data in 90 cases of primary gastrointestinal diffuse large B-cell lymphoma were analyzed by Kaplan-Meier method, Log-rank test and Cox regression model. Immunohistochemistry was carried out using EliVision and EnVision methods for CD20, CD3epsilon, CD10, bcl-6, MUM-1, CD5, bcl-2, GCET1, FOXP1, LMO2, BLIMP1 and Ki-67. RESULTS: The age of patients ranged from 27 to 83 years (mean = 58 years). The male-to-female ratio was 1.31:1. Amongst the 90 cases studied, 64.4% (58/90) involved the stomach and 35.6% (32/90) involved the intestine. The immunohistochemical findings were as follows: 100% positivity for CD20, 0% for CD3epsilon and CD5, 17.8% (16/90) for CD10, 75.6% (68/90) for bcl-6, 52.2% (47/90) for MUM-1 (cut off was 30%), 43.3% (39/90) for MUM-1 (cut off was 80%), 50.0% (45/90) for GCET1, 45.6% (41/90) for FOXP1, 23.3% (21/90) for LMO2, 42.2% (38/90) for bcl-2 and 8.9% (8/90) for BLIMP1. The Ki-67 index ranged from 20% to 95% (median = 80%). According to Hans algorithm, 51.1% of the cases belonged to germinal center B-cell (GCB) subtype and 48.9% belonged to non-GCB subtype. In contrast, Choi algorithm classified 55.6% cases as GCB subtype and 44.4% as activated B-cell (ABC) subtype. According to Tally algorithm, 34.4% were of GCB subtype and 65.6% of non-GCB subtype. Most of the patients (67.8%, 61/90) received chemotherapy and 68.9% (62/90) underwent surgical resection. The overall 2, 3 and 5-year survival rates were 58.5%, 52.8% and 49.8%, respectively. The overall 2, 3 and 5-year survival rates in the CHOP therapy group were 68.5%, 61.2% and 52.9%, respectively. CONCLUSIONS: There is no significant difference in ratio between the GCB and non-GCB/ABC subtypes by Hans and Choi algorithms. The non-GCB subtype seems to be more prevalent according to Tally algorithm. Although there is no significant difference in survival between GCB and non-GCB/ABC subtypes by the 3 algorithms, GCB subtype tends to show a better survival. In univariate analysis, LDH level, international prognostic index, chemotherapy, surgical resection, B symptoms, number of involved sites and clinical stage are found to have prognostic significance. In multivariate analysis, Choi algorithm, Tally algorithm, chemotherapy, surgical resection, LDH level and clinical stage are independent prognostic factors. PMID- 21615995 TI - [Clinicopathologic features of systemic EBV-positive T/NK-cell lymphoproliferative disease in adults]. AB - OBJECTIVE: To study the clinicopathologic features, immunophenotype, clonality and Epstein-Barr virus (EBV) status of systemic EBV-positive T/NK-cell lymphoproliferative disease in adults (ASEBV(+)T/NK-LPD). METHODS: Twenty cases of ASEBV(+)T/NK-LPD were analyzed retrospectively with histopathologic review, immunohistochemistry and in-situ hybridization for EBV-encoded RNA (EBER). The follow-up data were collected. RESULTS: There were altogether 15 males and 5 females. The median age of the patients was 34 years. The average duration from onset of symptoms to diagnosis was 8.7 months. Fever (18/20), hepatosplenomegaly (18/20) and lymphadenopathy (17/20) were the main clinical manifestations. Eleven of the 17 patients died during follow-up, with a mean survival of 2.9 months. Histologically, there was obvious expansion of T zone of the involved lymph nodes, associated with diminished lymphoid follicles. The interfollicular areas were widened and infiltrated by small to median-sized lymphoid cells which showed only mild atypia. Scattered large lymphoid cells were not uncommon. The nodal capsule was thickened in 6 cases. Focal necrosis was seen in 9 cases. Sinus histiocytic proliferation with erythrophagocytosis was observed in 3 cases. In addition, there were mild atypical lymphoid cells infiltrate into the liver, spleen, intestinal mucosa and bone marrow. Immunohistochemical study and in-situ hybridization showed that the EBER-positive cells were of T-cell lineage, with CD3 expression. They were also positive for cytotoxic molecules (granzyme B or TIA-1). Only 1 case was CD56 positive. A predominance of CD8-positive cells was demonstrated in 8 of the 14 cases studied, while CD4-positive cells predominated in the remaining 5 cases. One case showed similar proportion of CD8 and CD4 positive cells. The number of EBER-positive cells ranged from 30 to more than 300 per high-power fields. These EBER-positive cells were of small to large size and located mainly in the expanded T zone and occasionally in the germinal centers. Three of the 7 cases exhibited clonal rearrangement of T-cell receptor gamma gene, while the other 4 cases exhibited polyclonal rearrangement of T-cell receptor gamma gene. CONCLUSIONS: ASEBV(+)T/NK-LPD is a systemic disease with a subacute or chronic clinical course. Most patients suffer from relapsing fever, lymphadenopathy and hepatosplenomegaly. The disease is characterized by proliferation of EBV-infected cytotoxic T cells. The T zone of the involved lymph nodes shows expansion by mildly atypical lymphoid cells. The disease is associated with poor clinical outcome and can be life-threatening. The patients often die of multiorgan failure and bleeding. PMID- 21615996 TI - [Immunophenotypes and prognosis of diffuse large B-cell lymphoma: a study of 500 cases]. AB - OBJECTIVE: To study the immunophenotype and overall survival of diffuse large B cell lymphoma (DLBCL) classified according to the 2008 World Health Organization classification of tumors of hematopoietic and lymphoid tissues. METHODS: Five hundred cases of DLBCL were retrospectively analyzed with histologic review, immunohistochemistry, gene rearrangement study, in situ hybridization and fluorescence in situ hybridization. Follow-up data were collected. The overall survival rates of germinal center B-cell (GCB) and non-germinal center B-cell (non-GCB) subtypes, as well as those of DLBCL, not otherwise specified (NOS) and Epstein-Barr virus (EBV)-positive DLBCL of the elderly, were compared. RESULTS: DLBCL-NOS was the commonest subtype which accounted for 77.2% (386/500) of the cases. EBV-positive DLBCL of the elderly, primary DLBCL of central nervous system, primary mediastinal (thymic) large B-cell lymphoma and T cell/histiocyte rich large B-cell lymphoma accounted for 9.4% (47/500), 4.4% (22/500), 2.8% (14/500) and 2.6% (13/500), respectively. 68.5% (219/320) of DLBCL-NOS belonged to non-GCB subtype. The percentage of GCB subtype and CD5-positive subtype were 28.4% (91/320) and 3.1% (10/320), respectively. Comparison of the overall survival, GCB and non-GCB immunophenotypic groups have no significant difference (P = 0.93). And the same result in which of the EBV-positive DLBCL of the elderly and DLBCL-NOS group, before and after age matched (P = 0.13 and 0.28, respectively). A double-hit lymphoma was found by FISH detection, which presenting as gray zone lymphoma in morphology. CONCLUSIONS: By using Hans algorithm, GCB and non-GCB subtypes show no significant difference in overall survival. EBV-positive DLBCL of the elderly and DLBCL-NOS also do not have significant difference in overall survival. Fluorescence in situ hybridization technique is helpful in identification of DLBCL with rare phenotypes. PMID- 21615997 TI - [Perivascular epithelioid cell tumor, not otherwise specified: a clinicopathologic and immunohistochemical analysis of 31 cases]. AB - OBJECTIVE: To study the clinicopathologic characteristics of perivascular epithelioid cell tumor (PEComa), not otherwise specified (NOS) and to evaluate the diagnostic criteria for malignancy. METHODS: The clinical and pathologic features of 31 cases of PEComa-NOS were reviewed. The follow-up data available were analyzed. RESULTS: There were a total of 24 females and 7 males. The age of the patients ranged from 13 to 66 years (mean = 40 years). The site of tumor occurrence included gynecologic organs (n = 12), intraabdominal/peritoneal soft tissue (n = 10), gastrointestinal tract (n = 4), thigh (n = 2), mediastinum (n = 1), left groin (n = 1) and urinary bladder (n = 1). None of the cases was associated with tuberous sclerosis complex. Histologic examination showed that 23 cases (74%) were clear cell sugar tumor-like, 4 cases (13%) were clear cell myomelanocytic tumor-like and 4 cases (13%) were of mixed epithelioid-spindled morphology. According to the classification system proposed by Folpe et al, 19 cases (61%) were classified as malignant, 7 cases (23%) as PEComa of uncertain malignant potential and 5 cases (16%) as benign. The expression rates of HMB45, smooth muscle actin and desmin in tested cases were 100% (31/31), 67% (14/21) and 6/18, respectively. Follow-up data (1 to 56 months) were available in 23 cases (74%). Amongst the 16 cases of malignant PEComa, 7 patients were still alive with no evidence of disease, 6 patients were alive with unresectable or recurrent/metastatic disease and 3 patients died of the disease. The local recurrence and metastasis in those 16 cases were 6 cases and 5 cases, respectively. One of the 4 patients with PEComa of uncertain malignant potential died, while the remaining 3 patients and all of the patients with benign PEComa had an uneventful clinical course. CONCLUSIONS: The classification system of PEComas proposed by Folpe et al. is reliable in routine practice. Correlation with the clinical and radiologic findings however is prudent when dealing with core biopsy specimens or sampling from exploration laparotomy. Owing to the histologic heterogeneity of this entity, thorough understanding of the morphologic spectrum is essential in arriving at a correct diagnosis. PMID- 21615998 TI - [Establishment and characterization of a nude mice model of human diffuse large B cell lymphoma]. AB - OBJECTIVE: To establish a diffuse large B-cell lymphoma (DLBCL)-mice model using human DLBCL cell line LY8, to investigate its characteristics of growth and to provide a model for in vivo study of DLBCL pathogenesis and treatment. METHODS: LY8 cells were injected subcutaneously into the right flank of nude mice. Harvested tumor tissues were cut into small pieces of 1.5 mm * 1.5 mm * 1.5 mm and implanted subcutaneously into nude mice. Tumor growth was visualized and the histologic characteristics were documented. Expression of LCA, CD20, CD79alpha, Ki-67, CD3, CD45RO, bcl-6, MUM-1, CD10 and bcl-2 were examined by using immunohistochemistry. IgH clonal rearrangement and status of three microsatellite loci (D14S68, D18S69, D20S199) in the xenografted tumor samples and the parental cell line LY8 were detected using PCR amplification followed by PAGE. RESULTS: The subcutaneous xenograft DLBCL model was successfully established by using cell line LY8, and a stable growth was achieved up to the 9th generation. The tumor in each generation showed similar growth characteristics and the rate of subcutaneous tumor formation was 91.9% (114/124). The tumor growth was observed from the 2nd week after implantation, reaching 1.3 cm in major diameter at the 3rd week and 2.0 cm at the 4th week. The tumor had identical morphological characteristics with those of human DLBCL, and expressed LCA, CD20, CD79alpha, bcl-6, MUM-1, CD10 and bcl-2. The tumor of xenograft mice and cell line LY8 showed identical IgH rearrangement and microsatellite length. CONCLUSIONS: A human DLBCL bearing mouse model was successfully established. The mice model is similar to human counterpart with high stability and repeatability. Therefore, it provides an ideal animal model for in vivo studies of the biological characteristics and treatment of DLBCL. PMID- 21615999 TI - [Meningeal hemangiopericytoma: a clinicopathologic study of 17 cases]. PMID- 21616000 TI - [Expression and significance of CXCR7 in human colorectal tumor]. PMID- 21616001 TI - [Expression status of HER2 in mammary and extramammary Paget's disease]. PMID- 21616002 TI - [Chinese pathologic consensus for standard diagnosis of gastrointestinal and pancreatic neuroendocrine neoplasm]. PMID- 21616003 TI - [Evaluation and application of methylation-sensitive restriction enzymes-based quantitative PCR for DNA methylation detection]. PMID- 21616004 TI - [Preparation and application of three-dimensional pathology gross specimens]. PMID- 21616005 TI - [Granulomatous slack skin with anaplastic large cell lymphoma: report of a case]. PMID- 21616006 TI - [CD30-negative and ALK-positive anaplastic large cell lymphoma: report of a case]. PMID- 21616007 TI - [Primary diffuse large B-cell lymphoma in uterine corpus: report of a case]. PMID- 21616008 TI - [Congenital fibrosarcoma with diffuse metastases in fetus: report of a case]. PMID- 21616009 TI - [Mazabraud syndrome: report of a case]. PMID- 21616010 TI - [Laryngeal cryptococcosis: report of a case]. PMID- 21616011 TI - [Unclassified B-cell lymphomas with "grey zone" characteristics]. PMID- 21616012 TI - [Signaling pathways in pathogenesis of diffuse large B-cell lymphoma]. PMID- 21616013 TI - [Advances in research on EBV-positive T/NK cell lymphoproliferative disease]. PMID- 21616014 TI - The RATPAC (Randomised Assessment of Treatment using Panel Assay of Cardiac markers) trial: a randomised controlled trial of point-of-care cardiac markers in the emergency department. AB - OBJECTIVES: To evaluate the clinical effectiveness and cost-effectiveness of using a point-of-care cardiac marker panel in patients presenting to the emergency department (ED) with suspected but not proven acute myocardial infarction (AMI). DESIGN: Multicentre pragmatic open randomised controlled trial and economic evaluation. SETTING: Six acute hospital EDs in the UK. PARTICIPANTS: Adults presenting to hospital with chest pain due to suspected but not proven myocardial infarction, and no other potentially serious alternative pathology or comorbidity. INTERVENTIONS: Participants were allocated using an online randomisation system to receive either (1) diagnostic assessment using the point of-care biochemical marker panel or (2) conventional diagnostic assessment without the panel. All tests and treatments other than the panel were provided at the discretion of the clinician. MAIN OUTCOME MEASURES: The primary outcome was the proportion of patients successfully discharged home after ED assessment, defined as patients who had (1) either left the hospital or were awaiting transport home with a discharge decision having been made at 4 hours after initial presentation and (2) suffered no major adverse event (as defined below) during the following 3 months. Secondary outcomes included length of initial hospital stay and total inpatient days over 3 months, and major adverse events (death, non-fatal AMI, life-threatening arrhythmia, emergency revascularisation or hospitalisation for myocardial ischaemia). Economic analysis estimated mean costs and quality-adjusted life-years (QALYs), and then estimated the probability of cost-effectiveness assuming willingness to pay of L20,000 per QALY gained. RESULTS: We randomised 1132 participants to point of care and 1131 to standard care, and analysed 1125 and 1118, respectively [mean age 54.5 years, 1307/2243 (58%) male and 269/2243 (12%) with known coronary heart disease (CHD)]. In the point-of-care group 358/1125 (32%) were successfully discharged compared with 146/1118 (13%) in the standard-care group [odds ratio (OR) adjusted for age, gender and history of CHD 3.81; 95% confidence interval (CI) 3.01 to 4.82, p < 0.001]. Mean length of the initial hospital stay was 29.6 hours versus 31.8 hours (mean difference = 2.1 hours; 95% CI -3.7 to 8.0 hours, p = 0.462), while median length of initial hospital stay was 8.8 hours versus 14.2 hours (p < 0.001). More patients in the point-of-care group had no inpatient days recorded during follow up (54% vs 40%, p < 0.001), but mean inpatient days did not differ between the two groups (1.8 vs 1.7, p = 0.815). More patients in the point-of-care group were managed on coronary care [50/1125 (4%) vs 31/1118 (3%), p = 0.041]. There were 36 (3%) patients with major adverse events in the point-of-care group and 26 (2%) in the standard-care group (adjusted OR 1.31; 95% CI 0.78 to 2.20, p = 0.313). Mean costs per patient were L1217 with point-of-care versus L1006 with standard care (p = 0.056), while mean QALYs were 0.158 versus 0.161 (p = 0.250). The probability of standard care being dominant (i.e. cheaper and more effective) was 0.888. CONCLUSIONS: Point-of-care testing increases the proportion of patients successfully discharged home and reduces the median (but not mean) length of hospital stay. It is more expensive than standard care and unlikely to be considered cost-effective. TRIAL REGISTRATION: Current Controlled Trials ISRCTN37823923. FUNDING: This project was funded by the NIHR Health Technology Assessment programme and will be published in full in Health Technology Assessment; Vol. 15, No. 23. See the HTA programme website for further project information. PMID- 21616015 TI - Editorial Issue 1 2011: practical research to improve care. PMID- 21616016 TI - Prescribing for the elderly: ethical considerations. AB - This article reports on a few ethical considerations associated with prescribing for the elderly. A large number of elderly people are on several medications for the treatment of various chronic conditions that develop with age. Most of these treatments involve symptom management and prevention of symptoms likely to occur from untreated diseases. Serious consideration should be given when prescribing for the elderly. These include: allowing the elderly and their families to be involved in their treatment; social support; and cost-benefit considerations. A lack of clear therapeutic guidelines exists for prescribing in this group of people that is confounded by factors such as quality of life and life expectancy. PMID- 21616017 TI - Better knowledge, better health: piloting an education intervention in chronic condition self-management support. AB - Chronic condition self-management and lifestyle risk modification education is paramount for General Practice registrars. A multi-dimensional learning package 'Better Knowledge, Better Health' was developed and piloted to improve General Practice registrars' understanding of their role in supporting chronic condition self-management in patients with osteoarthritis. This pilot study was supported by the Australian Better Health Initiative. Pre-training learning needs analysis with a new intake of General Practice registrars (n=40) indicated high levels of confidence in supporting patients in chronic condition management and lifestyle risk modification, and locating and interacting with local resources and allied health professionals. Conversely, interviews with General Practice Supervisors (n=13) found most would not identify chronic condition self-management skills as priorities for registrar learning. Supervisors were also not familiar with core principles of chronic condition self-management, in particular application of motivational interviewing to behaviour change. Disparities between General Practice Supervisors' perceptions of the importance of chronic condition self management and lifestyle risk modification education and levels of access to learning opportunities in chronic condition self-management for registrars are discussed. Difficulties in implementing a pilot study within tight timeframes are also explored. PMID- 21616018 TI - Development of a nurse-led primary healthcare service for injecting drug users in inner-city Sydney. AB - Injecting drug users (IDUs) experience numerous health problems, but report barriers to utilising general practitioners (GPs). A nurse-led Harm Minimisation based Primary Healthcare (HMPH) service for IDUs was established within a needle and syringe program in inner-city Sydney with Area Health Service medical support and clinical governance. This paper aimed to describe the HMPH service, review service utilisation and assess nurses' perceptions of their work with IDUs. A review of the most recent 200 clinic files was undertaken. Service utilisation, GP and other health service use and access were extracted and analysed using SPSS. A semi-structured qualitative interview with clinic nurses regarding their experience working with IDUs and local GPs was conducted and analysed. Since its inception in mid-2006, the service has been utilised by 417 clients. Of the most recent 200 files, blood-borne virus and sexually transmitted infection screening were the primary reason for presentation (64.5%). At least one follow-up visit was attended by 90% of clients. A total of 62% of clients reported consulting a GP in the last 12 months. The service provided 102 referrals. Nurses believed that IDUs tend to utilise GPs ineffectively and that self-care is a low priority, but that they can support IDUs to overcome some barriers to GPs and facilitate access. Targeted primary health care services led by nurses with focussed medical support and co-located with needle and syringe programs can fill an important gap in delivering and facilitating health care to IDUs. PMID- 21616019 TI - Evaluation of a rural community pharmacy-based Waist Management Project: bringing the program to the people. AB - Time constraints and lack of awareness of risk factors for future chronic disease development prevent many young adults from accessing lifestyle programs offered by local health services. This study aimed to determine the effectiveness of a rural pharmacy-based multidisciplinary healthy lifestyle pilot program on reducing risk factors for chronic disease development among young adults. Individuals under the age of 50 with chronic disease risk factors were referred to the program. All subjects were provided with free after-hours nutritional counselling from a dietitian at the local community pharmacy, a comprehensive medication review conducted by the pharmacist, gym membership and access to cooking classes and supermarket tours. Selected participants also received bulk billed GP appointments and assistance with establishing a home vegetable garden. Body weight, waist circumference, fruit and vegetable consumption and physical activity were assessed at baseline and after program conclusion. Forty participants regularly attended the program between March 2009 and March 2010. At program conclusion, mean body weight was reduced by 3.8+/-6.7kg (P<0.001) and waist circumference reduced by 3.9+/-6.5cm (P<0.001). Fruit consumption increased by 1.2+/-0.2 serves/day (P<0.001) and vegetable intake increased by 1.6+/-1.0 serves/day (P<0.001). Participants also spent an average of 88.0+/-47.7 more min/week (P<0.001) engaged in physical activity on completion of the program. The community pharmacy provided an accessible location for the delivery of a successful chronic disease risk reduction program targeting young adults in a rural area. PMID- 21616020 TI - Physician's assistants: a workforce solution for Australia? AB - Significant medical workforce shortages, particularly in rural and remote locations, have prompted a range of responses in Australia at both state and Commonwealth levels. One such response was a pilot project to test the suitability of the Physician Assistant (PA) role in the Australian context. Five US-trained and accredited PAs were employed by Queensland Health and deployed in urban, rural and remote settings across Queensland. A concurrent mixed-method evaluation was conducted by Urbis, an independent research firm. The evaluation found that the PAs provided quality, safe clinical care under the supervision of local medical officers. The majority of nurses and doctors who worked with the PAs believed that the PAs made a positive contribution to the health care team by increasing capacity to meet patient needs; reducing on-call requirements for doctors; liaising with other clinical team members; streamlining procedures for efficient patient throughput; and providing continuity during periods of doctor changeover. The Pilot demonstrated that a delegated PA role can provide safe, quality health care by augmenting an established healthcare team. The PA role has the potential to benefit the community by increasing the capacity of the health care system, and to improve recruitment and retention by providing an additional professional pathway. The small size of the Pilot limits the ability to generalise regarding the future efficacy of the PA role in Australia. Further research is required to test training and deployment of PAs in a wider range of Australian clinical settings, including general practice and rural health clinics. PMID- 21616021 TI - Transgenerational learning: maximising resources, minimising teaching gaps and fostering progressive learning. AB - The need to rationalise teaching resources underpinned a project at Monash University that used a Delphi technique to re-examine the teaching curriculum of two key topic areas in the medical curriculum - ophthalmology and dermatology - from an undergraduate, graduate and vocational perspective. Using Bloom's taxonomy the learning objectives from these topic areas were collated and analysed. This process allowed the revising and redistributing of learning objectives of the curricula to reduce the likelihood of duplication of teaching or more importantly gaps in teaching occurring. This process highlighted the potential utility of using a transgenerational approach to curriculum planning but the outcomes are limited due to the small number of participating educators and the lack of formal evaluation of the method. PMID- 21616022 TI - A review of pharmacist recommendations in an aged care facility. AB - The aim of this study was to analyse the types of and rationale for the clinical recommendations made by the pharmacist in a rural aged care facility to improve patient safety. The classes of drugs associated with the pharmacist's recommendations were also identified. A related aim was to determine their degree of acceptance by medical practitioners. A retrospective, cross-sectional study design was used to review 56 aged care residents' case notes over a 12-month period. The main outcome measures included: the types of and reasons for recommendations made by the pharmacist; classes of drugs associated with the pharmacist's recommendations; and the implementation rate of the pharmacist's recommendations by the medical practitioner. A total of 196 recommendations were made by the pharmacist to the residents' existing medications. The main types of recommendations were alteration to residents' monitoring (49%), discontinuation of drug treatment (19%) followed by initiation of drug treatment (17%). The main reasons for the recommendations were to reduce potential side-effects (45%), symptom control (32%) and to increase drug efficacy (19%). Analysis of medical practitioners' case notes estimated that 70% of the pharmacist's recommendations were being implemented by the residents' medical practitioner. This case notes analysis reinforces the importance of doctor-pharmacist collaboration in the management of residents' medications in aged care facilities. PMID- 21616024 TI - Women's experience of becoming caregivers to their ill partners: Gadamerian hermeneutics. AB - The recent morbidity trends in Australia indicate a steep rise in the number of individuals living with chronic illness who rely on the assistance of mostly women carers. Consequently, supporting and promoting carers' health should be a priority to sustain their health, and ensure their ability to provide care to a significant other. This interpretive hermeneutic study explored the lived experience of women during the transition of becoming carers to their male partners with various health conditions. Gadamerian hermeneutics were used to research participants' experiences allowing for interpretations that incorporated understandings of both worlds, those of the researcher and the participants. This negotiation of understandings made it possible to create mutual and new knowledge. The phenomenon 'entering transition' revealed itself as a precarious balance between a sense of loss and coping with the life changing situation. Yearning to preserve that equilibrium, the women did not wish to be considered as carers to their partners but expressed a need for maintaining the cohesiveness of their identity as part of a couple. One of the recommendations arising out of this study is that a different approach is needed to assist nurses and other health professionals to support this specific population of women carers in the community and in hospital settings. PMID- 21616023 TI - Aspects of work-life balance of Australian general practitioners: determinants and possible consequences. AB - The Australian general practitioner (GP) workforce, especially younger generation GPs and female GPs, increasingly prioritises work-life balance (WLB). Good WLB is associated with decreased interest of medical students in general practice as a speciality choice as well as good health and wellbeing, and decisions of GPs to retire early. Therefore, understanding the role played by different factors in achieving WLB is crucial to ensure a sufficient GP workforce necessary to meet the rising demands of health care. There is a dearth of empirical, quantitative, large, population-based studies assessing the level of WLB in the Australian GP population as well as contributing and consequent factors. Our study fills this identified gap in the current literature. This study aimed to investigate the extent, determinants and possible consequences of WLB of Australian GPs. Data for this study come from the baseline cohort of the Medicine in Australia: Balancing Employment and Life (MABEL) longitudinal, population-level survey. Questionnaires tailored specifically for GPs and GP registrars were sent to all clinically active GPs registered in the Australian Medical Publishing Co. (AMPCo) database (n=22137), with a choice of completing either a paper or online version. Data were collected between June and December 2008. STATA (10.0) was used for conducting weighted data analyses. Regression methods were applied for assessing the associations between dependent and independent variables. Of the 3906 GPs (17.6%) who responded, 53% reported that the balance between their personal and professional commitments was about right. Generation X GPs and females reported a better WLB than baby boomers and males respectively. However, those reporting good WLB also worked significantly fewer hours than those reporting poor WLB. GPs who reported good opportunities for leisure activities and perceived that they have good health also reported better WLB. Contrastingly, those reporting difficulty in taking time off when wanted, working unpredictable and longer hours reported poor WLB. Importantly, poor WLB was associated with increased intention to reduce total hours worked by GPs. Several factors relating to family and social circumstances were not significantly associated with WLB of GPs in this study. Only half of the GP workforce reported good WLB. Flexibility of work hours, opportunities for leisure activities and good health of GPs have the potential to enhance the WLB and hence promote greater GP participation in the workforce. PMID- 21616025 TI - 'Adolescence is difficult, some kids are difficult': general practitioner perceptions of working with young people. AB - General practitioners (GPs) are the health care providers from which young people are most likely to seek help. However, the rate at which young people access GPs is less than ideal. Four focus groups were conducted with groups of GPs in New South Wales to inform the development of a GP training program on youth health. Analysis of the focus group interviews yielded three themes that describe the context in which GPs work with young people, how GPs see young people and their work with them, and GPs' expressed training preferences. GPs described working with young people as difficult, which in turn suggests that the self-efficacy of GPs may need to be addressed in training and resource development. GPs also described systemic barriers to working with young people that can be addressed through training, advocacy and policy development. PMID- 21616026 TI - Childhood obesity: how do Australian general practitioners feel about managing this growing health problem? AB - General practitioners (GPs) are ideally placed to identify and treat childhood obesity, but its prevalence continues to rise and evidence for effective GP interventions is lacking. Further analysis of the barriers to effective identification and management of childhood obesity is warranted. This survey aimed to explore how Queensland GPs feel about managing the growing problem of childhood obesity. A cross-sectional survey was sent to a random sample of 573 Queensland GPs about perceptions of diagnosis and management of childhood obesity. A total of 30% of GPs responded (n=170). The main perceived obstacles to identification of childhood obesity were uncertainty about definition criteria and how to calculate body mass index, and lack of access to body mass index percentile charts. The main perceived obstacles in managing childhood obesity were lack of financial incentive, time constraints, lack of health system support and parental resistance. Only 22% of respondents indicated awareness of the National Health and Medical Research Council guidelines for management of obese children and 92% had never used any formal clinical guidelines in assessment or management of childhood obesity. Addressing these barriers to identification of childhood obesity by GPs may facilitate more effective management. Strategies include greater emphasis on this issue in general practice training, financial incentives for diagnosis and management, incorporating clinical management guidelines into medical software, and increasing allied and community health support. PMID- 21616027 TI - Health status and use of health services among recently arrived men with refugee backgrounds: a comparative analysis of urban and regional settlement in South east Queensland. AB - Approximately one-third of refugee and humanitarian entrants to Australia are adult men. Many of these men and their families settle in regional areas. Little is known about the health status of refugee men and the use of health services, and whether or not there are differences between those living in urban and regional areas. This paper reports on the cross-sectional differences in health status and use of health services among a group of 233 recently arrived refugee men living in urban and regional areas of South-east Queensland. Overall, participants reported good levels of subjective health status, moderate to good levels of well-being, and low prevalence of mental illness. Men living in urban areas were more likely to have a long-standing illness and report poorer health status than those settled in regional areas. In contrast, men living in regional areas reported poorer levels of well-being in the environment domain and were more likely to visit hospital emergency departments. Targeted health promotion programs will ensure that refugee men remain healthy and develop their full potential as members of the Australian community. Programs that facilitate refugees' access to primary health care in regional areas may promote more appropriate use of hospital emergency departments by these communities. PMID- 21616028 TI - Practice-based chronic condition care coordination: challenges and opportunities. AB - This paper explores issues related to the types of support that practice nurses require to engage in care coordination for people with chronic conditions. A sample of practice nurses and general practitioners participated in a focus group discussion to identify their perspectives on the role of practice nurses as providers of care coordination, the specific tasks that might be conducted and the types of support that might be required. The data were analysed using thematic analysis. The findings suggested there was considerable confusion about care coordination and a lack of conceptual clarity. Nevertheless, nurses were committed to engaging in activities that would promote care coordination. Four main themes emerged that indicated the importance of a developmental and well supported implementation process. These themes included the need for cultural change within the whole practice, increased capacity to develop trusted and tested partnerships, appropriate role definition and a full understanding of the financial models that could support care coordination. Practice nurses clearly have a role in care coordination, but models of care coordination need to be localised and contextualised within specific GP practices. Cultural change will, in many instances, be critical to the development of localised programs. Broad supportive structures, including ongoing mentorship and administrative assistance (particularly with financial and procedural aspects of care coordination) will be required when implementing programs that enhance roles for practice nurses. PMID- 21616029 TI - Factors that influence physical activity for pregnant and postpartum women and implications for primary care. AB - Many pregnant women and women of child-bearing age do not engage in the recommended levels of physical activity despite the well known benefits. Pregnancy and the postpartum period can be a time when inactivity actually increases. Women who experience gestational diabetes mellitus (GDM) during their pregnancy are often advised to become more active in order to ameliorate their increased risk of developing type 2 diabetes. Health professionals have an influential role in promoting physical activity, which would be enhanced with an understanding of the factors that positively and negatively influence women's participation in physical activity during pregnancy and in the postpartum period. This research sought to explore these factors with pregnant and postpartum women including those who had experienced GDM and the attention given to physical activity during pregnancy. A survey was developed after a critical review of factors identified from previous studies. Women were recruited from the antenatal clinic, community health centres and the local media. Results from 72 women are reported from a predominately well educated, Caucasian population. Overall, the results were confirmatory of factors previously identified. Lack of child care, time constraints, no time and feeling unwell during pregnancy hindered activity and factors that facilitated activity included family support, enjoyment of activity and to prevent later health problems. It was also found that non-GDM women are given minimal advice about exercise during pregnancy. A checklist has been developed for health professionals, in partnership with women, to direct attention to the factors that enable and hinder participation in physical activity during and after pregnancy. PMID- 21616030 TI - Walking for fitness: is it enough to maintain both heart and bone health? AB - Exercising at levels of whole body accelerations exceeding 3.6g has been shown to have positive effects on cardiovascular fitness, bone density and balance. This pilot research project evaluated the whole body accelerations and cardiovascular challenge provided by selected walks in the Canberra region of Australia to determine if walks could be ranked according to potential level of impact on both cardiovascular fitness and bone health. Nine participants, who described themselves as walking at least 3km, three times per week, wore a data logging device recording heart rate, acceleration and GPS position while walking three outdoor tracks: (1) the running track of an athletics stadium; (2) on a hill climb path through bushland; and (3) on a route through suburban streets. There was a significant difference (P<0.05) for heart rate, distribution of whole body accelerations and average walking speed between track 2 and tracks 1 and 3. There was a significant difference for heart rate, distribution of whole body accelerations and average walking speed between the walks. The running track and the suburban walk provide a moderate exercise challenge, with the hill climb walk providing progressively greater vertical height challenge, resulting in an increased cardiovascular exercise challenge. No participant effectively exceeded the threshold for achieving a positive impact on bone density (100 or more accelerations/day >3.6g) on track 1, and only two of the nine participants intermittently achieved this threshold on tracks 2 and 3. PMID- 21616031 TI - Dementia: opportunities for risk reduction and early detection in general practice. AB - This project aimed to measure general practitioner (GP), practice nurse and patient health literacy about memory problems, dementia and its risk factors. Data were collected from general practices across Australia and a smaller sample in England. Questionnaires explored sources and adequacy of dementia knowledge and a randomised controlled trial tested the intervention of a dementia risk reduction pamphlet on patient knowledge of dementia risk reduction strategies. Data were analysed using SPSS software. The results of 621 questionnaires from patients aged over 30 years showed 37% had memory concerns, 6% recalled having a memory test, 52% would like a memory test and 15% had heard about dementia from their GP. Patients receiving the intervention were significantly more likely to be aware of dementia risk reduction strategies (P<=0.005). The results of 153 GP/nurse questionnaires indicated 64% thought a doctor should discuss dementia with patients despite only 21% assessing their dementia knowledge as adequate. There was no significant difference in responses between Australia and England (P>=0.05). The frequency of documentation of Mini-Mental State Examination and dementia diagnosis in computerised medical records of patients over 75 years was less than 0.01. These results demonstrate that many adult patients attending GPs have memory concerns, associate dementia with memory loss, and are receptive to information about dementia risk reduction. Most general practitioners and their nurses rate their dementia knowledge as inadequate with few testing for memory problems or discussing dementia with their patients. PMID- 21616032 TI - Enhancing palliative care in rural Australia: the residential aged care setting. AB - The delivery of palliative care in residential aged care communities is challenging, even more so in rural areas due to workforce ageing and shortages. The objectives of the present study were to: (i) assess the needs of, and quality of palliative care delivered to residents of 16 residential aged care facilities in rural southern Australia; and (ii) identify the needs of care staff to facilitate the delivery of quality palliative care. A cross-sectional survey of all residents, assessing the degree of functional limitation, stage of palliative care, and the presence of several quality indicators was conducted. Separate focus groups of care staff and relatives of residents sought information on the quality of care delivered, perceived strengths and weaknesses of the care delivered, and education and training needs. Quality palliative care in residential aged care facilities (RACFs) is hampered by workforce shortages, with low ratios of registered nurses, limited access to general practitioners after hours, and some communication difficulties. Some staff reported low confidence in technical and psychosocial aspects of care, especially for relatives. Relatives described mostly appropriate care, while acknowledging workload constraints. Most residents whose condition was unstable, deteriorating or terminal received advance care planning, though family expectations and unwillingness to discuss end-of-life care did tend to delay planning. Unstable residents with a reasonable prognosis were more likely to be transferred to hospital than terminally ill residents. Palliative care in participating RACFs appears to be adequate. Provision of targeted education for health care providers and implementation of protocols for advance care planning and end-of life care pathways will enhance this care. PMID- 21616033 TI - The qualitative interview and challenges for clinicians undertaking research: a personal reflection. AB - Drawing on my doctoral experience the aim of this article is to present my transition from practitioner to novice researcher and the challenges I encountered when undertaking qualitative in-depth interviews. The contents of my research diary were coded for words, sentences and paragraphs and were then grouped into themes and subsequently organised into concepts and categories. The analysis identified one core category: 'changing states: learning to become a researcher'. The related categories included 'guessing responses', 'confusing boundaries' and 'revealing hidden concepts'. These concepts provide a description of how I learnt to become a researcher and became a changed state. The paper provides practitioners with practical examples of my transition from practitioner to novice researcher. I offer some tips for practitioners who wish to undertake research in their clinical role. PMID- 21616034 TI - Rural research capacity building program: capacity building outcomes. AB - The Rural Research Capacity Building Program commenced in 2006 with the aim of developing research skills in rural health workers. The program was based on the capacity building principles of workforce development, organisational development, resource allocation, partnership and leadership. Qualitative methods were used to assess capacity building outcomes. A sample of candidates from the 2006 and 2007 cohorts were selected for interview using stratified random sampling and supplemental purposive sampling. Twenty-five individual semi structured interviews were conducted with candidates, their managers and mentors. Interviews were thematically analysed. The program components of teaching, mentoring and networking led to the development of research skills in candidates undertaking the program. This workforce development resulted in workplace change, particularly where the candidate's project was 'close to practice' and they had management support. The leadership shown and partnerships developed by the program managers enhanced the workforce development and organisational change outcomes. Resources, such as backfill and incidentals, were useful for candidates, but practicalities, such as availability of replacement staff, limited effectiveness. This study showed the value of using a capacity building framework and demonstrated that undertaking research on a topic close to practice positioned candidates to drive change within their organisation. PMID- 21616035 TI - Researcher development program of the primary health care research, evaluation and development strategy. AB - The Research Development Program (RDP) was initiated in 2004 under the Primary Health Care Research, Evaluation and Development (PHCRED) Strategy to increase the number and range of people with knowledge and skills in primary health care research and evaluation. RDP Fellows were invited to participate in an online survey about the effect the program had on their research knowledge, attitudes and practice. The response rate was 42% (105/248). Most were female (88%) with 66% aged between 31 and 50 years. Over two-thirds (72%) were health practitioners. Activities undertaken during the RDP ranged from literature reviews, developing a research question, preparing ethics submissions, attending and presenting at conferences and seminars, preparing papers and reports, and submitting grant applications. Despite the fact that only 52% agreed that the RDP time was adequate, 94% agreed that the RDP was a valuable experience, with 89% expressing interest in undertaking further research. These results indicate that this program has had a positive effect on the RDP Fellows in terms of their knowledge about research, their attitude to research, and the way they use research in their work. PMID- 21616038 TI - Roles of autoantibodies in central nervous system injury. AB - Stroke, traumatic brain injury (TBI), and spinal cord injury (SCI) cause irreversible damage to the nervous system. Although these are neurological disorders, pathology and loss of function also occur outside the nervous system and are often not easily explained by paralysis or impaired neural function. Emerging data indicate that much of the pathological sequelae that accompanies CNS trauma has characteristics of a self-directed immunological disease. Here, we outline those data, describing basic mechanisms of B cell activation and autoantibody synthesis after CNS injury. A summary of the anti-CNS autoantibodies that have been identified in humans and animals is provided along with a discussion of how autoantibodies may affect survival of neuronal and non-neuronal tissues and whether autoimmune reactions are feasible therapeutic targets after CNS trauma. PMID- 21616037 TI - HIV and bone metabolism. AB - The skeleton is an organ whose integrity is maintained by constant lifelong renewal involving coordinated removal of worn bone by osteoclasts and resynthesis of new bone by osteoblasts. In young adult humans and animals this process is homeostatic with no net gain or loss of bone mass. With natural aging and exacerbated by numerous pathological conditions, bone removal exceeds bone formation, disrupting homeostasis and resulting in bone loss. Over time, skeletal decline reaches clinical significance with development of osteopenia and eventually osteoporosis, conditions that dramatically increase bone fragility and the risk of fracture. Bone fractures can be devastating with significant morbidity and mortality. Over the last decade, it has become clear that skeletal renewal is strongly influenced by the immune system, a consequence of deep integration and centralization of common cell types and cytokine mediators, which we have termed the "immuno-skeletal interface." Consequently, dysregulated skeletal renewal and bone loss is a common feature of inflammatory conditions associated with immune activation. Interestingly, bone loss is also associated with conditions of immunodeficiency, including infection by the human immunodeficiency virus (HIV) that leads to acquired immunodeficiency syndrome (AIDS). Disruptions to the immuno-skeletal interface drive skeletal deterioration contributing to a high rate of bone fracture in HIV infection. This review examines current knowledge concerning the prevalence and etiology of skeletal complications in HIV infection, the effect of antiretroviral therapies (ART) on the skeleton, and how disruption of the immuno-skeletal interface may underlie bone loss in HIV infection and ART. PMID- 21616039 TI - Immune mechanisms in atherosclerosis and potential for an atherosclerosis vaccine. AB - A large body of evidence implicates the immune system in the pathogenesis and modulation of atherosclerosis. Dendritic cells and lymphocytes are among the many components of the immune system that are involved in modulating atherogenesis. This review focuses on the current knowledge of the complex role of the dendritic cells and lymphocytes in atherogenesis and the potential for immune-modulation therapies for atherosclerosis. PMID- 21616040 TI - The potential role of Th17 in mediating the transition from acute to chronic autoimmune inflammation: rheumatoid arthritis as a model. AB - T helper 17 cells (Th17) have arisen in the last 15 years as major effector cells in several chronic inflammatory states. In synovitis associated with rheumatoid arthritis (RA), Th17 emerged as being involved in driving the active acute phases and correlated with local and systemic parameters of inflammation; in particular, TCRzeta(dim) Th17 appear to be the greatest producers of IL-17 at the single-cell level. IL-1beta and IL-6, along with IL-23, arose as the major drivers of differentiation and local development of Th17, while IL-15 and cell-cell contact can trigger the local production of IL-17. TNF-alpha inhibition can reversibly block the migration of pathogenic effector memory TCRzeta(dim) T cells and CCR6+ Th17 from peripheral blood to inflamed tissues. IL-17 is a potent chemoattractant for pre-committed CD4+ T cells and neutrophils, and may promote the migration of B cells to lymphoid follicles in the chronic phase of synovial inflammation. Importantly, IL-17 drives osteoclastogenesis and neoangiogenesis in the RA joint. These data strongly suggest that Th17 are key effector cells in driving the transition from the acute to the chronic phase of RA inflammation. PMID- 21616041 TI - Diagnosis and treatment of irritable bowel syndrome. AB - Irritable bowel syndrome (IBS) is a chronic functional disorder of the gastrointestinal tract. The exact cause is unknown. The diagnosis should be made on clinical grounds, using symptom-based criteria such as the Manning or Rome criteria, unless symptoms are thought to be atypical. Excluding celiac disease in all patients consulting with symptoms suggestive of IBS is worthwhile, but evidence for performing other investigations to exclude organic disease is not convincing. No medical therapy for IBS has been shown to alter the disease course, and treatment has traditionally been directed towards symptom relief. The aim should be to improve the predominant symptom reported by the patient. Fiber, peppermint oil, or antispasmodic agents are beneficial as first-line therapies in some patients. Where these fail, emerging data have confirmed the efficacy of antidepressants, drugs acting on the 5-hydroxytryptamine receptor, and probiotics in the short-term treatment of IBS. There are a number of novel therapies under development that show promise, including non-absorbable antibiotics, lubiprostone, and linaclotide. This article will provide a summary of diagnostic criteria for IBS, evidence to support investigations to exclude organic disease, and current and emerging therapies in this field. PMID- 21616042 TI - Molecular imaging using positron emission tomography in colorectal cancer. AB - Positron emission tomography (PET) is one of the most important recent advances in cancer imaging. Molecular imaging using PET is now an integral part of multidisciplinary cancer care. In this review, the role of PET in colorectal cancer (CRC) is discussed, including its well established role in the assessment of recurrent disease and emerging applications such as initial staging, monitoring therapy efficacy, and radiotherapy planning. The development of new hybrid devices such as PET-magnetic resonance imaging along with the use of novel molecular probes in targeting specific pathways in oncogenesis will further improve patient management. PMID- 21616043 TI - Autoimmune encephalitis -- new awareness, challenging questions. AB - The field of autoimmune encephalopathies has expanded rapidly in the last few years. It is now well-established that a substantial proportion of encephalitides are associated with autoantibodies directed against the extracellular domains of cell-surface proteins which are critical in the regulation of neuronal excitability. These include LGI1, CASPR2, contactin-2 (VGKC-complex antibodies), and the NMDA, AMPA, and GABA(B) receptors. The clinical importance of these conditions lies in their frequent immunotherapy-response and, less commonly, their association with distinctive tumors. Studies which have examined cohorts of patients defined by these serum antibodies have identified a number of clinical features that have helped understand the core phenotypes of these conditions. In addition, sensitive antibody assays have allowed the expansion of the phenotypes to include a minority of patients with isolated epilepsies or psychoses. There is also evidence that autoimmune encephalitis may progress to adult-onset hippocampal sclerosis. Clinical, and accumulating scientific, data strongly suggest direct pathogenicity of these autoantibodies. The generation of the autoantibody, in some patients, can be explained by the presence of tumors which express their antigenic target. Serum antibody levels are higher than their levels in CSF in the vast majority of cases. However, the majority of patients do not harbor a tumor and the etiology of the disease in these patients is less clear. Below, we suggest models for the etiology and pathogenic mechanisms of these autoantibodies by incorporating concepts such as serum generation of the autoantibodies, the blood-brain barrier, intrathecal antibody production, and prodromal infections. PMID- 21616044 TI - Cytokines in the pathogenesis of ulcerative colitis. AB - Ulcerative colitis is a chronic inflammatory condition of the large intestine whose etiology remains largely unknown. Its pathogenesis involves the breakdown of intestinal mucosal homeostasis due to a genetically determined miscommunication between commensal flora and the gut associated immune system. Cytokines are central components of the inflammatory pathways that take place during the active and chronic phases of ulcerative colitis. Recent research has identified several novel cytokine systems that are upregulated at the mucosa of patients with ulcerative colitis and started to unveil their functional importance for disease pathogenesis. The significance of interleukin-13 (IL-13), TNF-like cytokine 1A (TL1A), IL-33, and their receptors in ulcerative colitis is strongly supported by converging expression and functional data. These molecular systems may define subgroups of patients with uniform immunological profiles. Within these subpopulations such novel cytokine systems may serve as markers of biological activity of the disease. More importantly, they may offer unique therapeutic opportunities through the development of drugs that specifically target and neutralize well-defined inflammatory pathways. PMID- 21616045 TI - Pharmacogenetic mechanisms underlying unanticipated drug responses. AB - Because most medicines have not been encountered by individuals of our species prior to treatment, it follows that treatment could uncover a previously silent genetic predisposition or could interact with a known genetic variation(s) to produce an unintended outcome. Pharmacogenetics encompasses the discovery, testing, and application of genetic variation as applied to therapeutic treatment and outcome. Two broad divisions of pharmacogenetics are recognized: pharmacokinetics, which describes genetic variations that affect drug metabolism, and pharmacodynamics which describes similar processes that have effects on drug targets, including downstream signaling pathways. The genetic mechanisms that underlie an altered drug response recapitulates most known sources of genomic variation. The most commonly encountered is sequence variation. This includes changes in the primary nucleotide sequence of coding, regulatory, and splice regions of a gene, the product of which affects, or is affected by, a drug. Less common forms of variability in the structure and function of the genome have also been found to underlie an individualized response to medicines. Among these are sequence variation in microRNA (miRNA) binding sites, which affects the ability of miRNA to regulate translation; pharmacoepigenetics, which examines heritable chromatin modifications; and copy number variation. Among the 158 currently registered pharmacogenetic clinical trials, the most frequent conditions or disease processes being studied are cancer, psychiatric disorders, and coagulation/ thrombosis. From this observation, it is postulated that pharmacogenetics has its greatest potential for optimizing the use of drugs with a high rate of failure or adverse outcomes. PMID- 21616046 TI - WHO urges polio-endemic countries to completely halt the transmission of the wild polio virus by 2012. PMID- 21616047 TI - Long term trends introduce a potential bias when evaluating the impact of the pneumococcal conjugate vaccination programme in England and Wales. AB - A pneumococcal conjugate vaccine (PCV7) was introduced into the United Kingdom's childhood immunisation schedule in September 2006. Evaluation of its impact on the incidence of invasive pneumococcal disease (IPD) as assessed by routine reports of laboratory-confirmed cases should take into account possible long-term trends due to factors like changes in case ascertainment. To this end, we compared pre-PCV7 trends in reported IPD incidence in England and Wales identified by blood culture with those for two other bacteraemias, Escherichia coli and non -pyogenic streptococci, for which there has not been any public health intervention. While no trend was detected in the age group 65 years and older, there was an annual increase of 3% and 11% in those aged under five years and between five and 64 years, respectively, which was similar for IPD and the other two pathogens. After PCV7 introduction, a continuing trend was only found for non-pyogenic streptococci in under five year-olds. These trends in the incidence for bacteraemias for which there has been no intervention could suggest that there have been changes in case ascertainment because of increased reporting or blood culturing. Accounting for them will improve the evaluation of the impact of PCV7 on IPD. PMID- 21616048 TI - Toxin producing Vibrio cholerae O75 outbreak, United States, March to April 2011. AB - The Florida Department of Health, Florida, United States, is investigating a Vibrio cholerae O75 outbreak. Ten cases with disease onsets from 23 March to 13 April 2011, presented with gastrointestinal symptoms of diarrhoea, nausea, vomiting, cramps, chills, and/or fever, after consuming raw or lightly cooked oysters harvested from Apalachicola Bay, Florida. Symptoms were milder than those during outbreaks of epidemic (serogroup O1 and O139) Vibrio cholerae; no case required rehydration treatment or hospitalisation. PMID- 21616049 TI - Influenza A(H1N1)2009 antibody seroprevalence in Scotland following the 2010/11 influenza season. AB - Following the 2010/11 influenza season, we determined the age- and location specific seroprevalence of antibodies against the influenza A(H1N1)2009 virus in Scotland. Samples were analysed by microneutralisation assay. Age/seropositivity profiles varied significantly between cities. The increases in seroprevalence relative to the previous influenza season (2009/10) were similar across age groups and geographic locations. However, the increased seropositivity in older adults appeared to be driven by exposure to vaccination, indicating significantly lower levels of infection than in younger age groups. PMID- 21616050 TI - A cluster of Listeria monocytogenes infections in hospitalised adults, Midlands, England, February 2011. AB - Hospital-acquired listeriosis cases are not commonly reported but remain a significant public health problem. We report on three cases in patients with underlying conditions occurring during one week in February 2011. The cases had common exposure to pre-packed sandwiches and salads manufactured in compliance with regulations. Breaches in cold chain and shelf life controls at hospital level were identified as key contributing factors. Rigorous hospital food management systems remain important for patient safety. PMID- 21616051 TI - Intracellular regulation of heterotrimeric G-protein signaling modulates vascular smooth muscle cell contraction. AB - The contractile function of vascular smooth muscle cells within the media of resistance arterioles is tightly connected to the role of these blood vessels in the maintenance of blood pressure homeostasis. Thus, much effort has been made to understand the intracellular signaling pathways that control vascular smooth muscle cell contractility with the aim that this knowledge will provide important clues for reducing the impact of uncontrolled blood pressure in our society. A key set of surface receptors, the G-protein coupled receptors, has been widely associated with the regulation of vascular smooth muscle cell contractility. Indeed, many of the current treatments for hypertension involve selective inhibition of these receptors. More recently, we have begun to understand the cellular mechanisms whereby G-protein coupled pathways are connected to the contractile machinery of the vascular smooth muscle cells. What has emerged is a view where there are multiple intracellular control points for G-protein signaling that coordinate and focus the extracellular stimuli into meaningful physiologic responses. This work will examine some of the recent advances in our understanding of G-protein signaling and its regulation of contractile function in vascular smooth muscle cells. PMID- 21616052 TI - Bis-THF motif of acetogenin binds to the third matrix-side loop of ND1 subunit in mitochondrial NADH-ubiquinone oxidoreductase. AB - Natural acetogenins are among the most potent inhibitors of bovine heart mitochondrial NADH-ubiquinone oxidoreductase (complex I). Our photoaffinity labeling study suggested that the hydroxylated bis-THF ring moiety of acetogenins binds at "site A" in the third matrix-side loop connecting the fifth and sixth transmembrane helices in the ND1 subunit [Kakutani et al. (2010) Biochemistry 49, 4794-4803]. Nevertheless, since this proposition was led using a photoreactive Deltalac-acetogenin derivative, it needs to be directly verified using a natural acetogenin-type probe. We therefore conducted photoaffinity labeling using a photoreactive natural acetogenin mimic ([(125)I]diazinylated natural acetogenin, [(125)I]DANA), which has a small photolabile diazirine group, in place of a hydroxy group, attached to the bis-THF ring moiety. Analysis of the photocross linked protein in bovine heart submitochondrial particles unambiguously revealed that [(125)I]DANA binds to the membrane subunit ND1 with high specificity. The photocross-linking was completely blocked in the presence of just a 5-fold excess of bullatacin, indicating that [(125)I]DANA is an excellent mimic of natural acetogenins and hence binds to the site that accommodates natural products. Careful examination of the fragmentation patterns of the cross-linked ND1 generated by different proteases and their combinations indicated that the cross linked residue is predominantly located at the supposed site A in the third matrix-side loop. PMID- 21616053 TI - Functional domain size in aggregates of light-harvesting complex II and thylakoid membranes. AB - The functional domain size for efficient excited singlet state quenching was studied in artificial aggregates of the main light-harvesting complex II (LHCIIb) from spinach and in native thylakoid membranes by picosecond time-resolved fluorescence spectroscopy and quantum yield measurements. The domain size was estimated from the efficiency of added exogenous singlet excitation quenchers phenyl-p-benzoquinone (PPQ) and dinitrobenzene (DNB). The mean fluorescence lifetimes tau(av) were quantified for a range of quencher concentrations. Applying the Stern-Volmer formalism, apparent quenching rate constants k(q) were determined from the dependencies on quencher concentration of the ratio tau(0)(av)/tau(av), where tau(0)(av) is the average fluorescence lifetime of the sample without addition of an exogenous quencher. The functional domain size was gathered from the ratio k(q)'/k(q), i.e., the apparent quenching rate constants determined in aggregates (or membranes), k(q)', and in detergent-solubilised LHCII trimers, k(q), respectively. In LHCII macroaggregates, the resulting values for the domain size were 15-30 LHCII trimers. In native thylakoid membranes the domain size was equivalent to 12-24 LHCII trimers, corresponding to 500-1000 chlorophylls. Virtually the same results were obtained when membranes were suspended in buffers promoting either membrane stacking or destacking. These domain sizes are orders of magnitude smaller than the number of physically connected pigment-protein complexes. Therefore our results imply that the physical size of an antenna system beyond the numbers of a functional domain size has little or no effect on improving the light-harvesting efficiency. PMID- 21616054 TI - The effect of long-chain bases on polysialic acid-mediated membrane interactions. AB - Negatively-charged polysialic acid (polySia) chains are usually membrane-bound and are often expressed on the surface of neuroinvasive bacterial cells, neural cells, and tumor cells. PolySia can mediate both repulsive and attractive cis interactions between membrane components, and trans interactions between membranes. Positively-charged long-chain bases are widely present in cells, are often localized in membranes and can function as bioactive lipids. Here we use Langmuir monolayer technique, fluorescence spectroscopy and electron microscopy of lipid vesicles to study the role of a simple long-chain base, octadecylamine (ODA), in both cis and trans interactions mediated by polySia in model membranes composed of ODA and dioleoylphospatidycholine (DOPC). When added free to an aqueous solution, polySia increases the collapse pressure of ODA/DOPC monolayers, reduces the effect of ODA on the limiting molecular area, inverses the values of excess area per molecule and of excess free energy of mixing from positive to negative, and induces fusion of ODA/DOPC vesicles. These results suggest that a polySia chain can act as a multi-bridge that mediates cis interactions between different components of a lipid membrane, disrupts membrane aggregates, and mediates trans interactions between lipids in apposing membranes. These observations imply that polySia in cellular systems can act in a similar way. PMID- 21616055 TI - Analysis of AcrB and AcrB/DARPin ligand complexes by LILBID MS. AB - The AcrA/AcrB/TolC complex is responsible for intrinsic multidrug resistance (MDR) in Escherichia coli. Together with the periplasmic adaptor protein AcrA and the outer membrane channel TolC, the inner membrane component AcrB forms an efflux complex that spans both the inner and outer membrane and bridges the periplasm of the Gram-negative cell. Within the entire tripartite complex, homotrimeric AcrB plays a central role in energy transduction and substrate selection. In vitro selected designed ankyrin repeat proteins (DARPin) that specifically bind to the periplasmic domain of AcrB were shown to ameliorate diffraction resolution of AcrB/DARPin protein co-crystals (G. Sennhauser, P. Amstutz, C. Briand, O. Storchenegger, M.G. Grutter, Drug export pathway of multidrug exporter AcrB revealed by DARPin inhibitors, PLoS Biol 5 (2007) e7). Structural analysis by X-ray crystallography revealed that 2 DARPin molecules were bound to the trimeric AcrB wildtype protein in the crystal, whereas the V612F and G616N AcrB variant crystal structures show 3 DARPin molecules bound to the trimer. These specific stoichiometric differences were analyzed in solution via densitometry after microchannel electrophoresis, analytical ultracentrifugation and via laser-induced liquid bead ion desorption mass spectrometry (LILBID-MS). Using the latter technology, we investigated the gradual disassembly of the AcrB trimer and bound DARPin ligands in dependence on laser intensity in solution. At low laser intensity, the release of the detergent molecule micelle from the AcrB/DARPin complex was observed. By increasing laser intensity, dimeric and monomeric AcrB species with bound DARPin molecules were detected showing the high affinity binding of DARPin to monomeric AcrB species. High laser intensity LILBID MS experiments indicated a spectral shift of the monomeric AcrB peak of 3.1kDa, representing a low molecular weight ligand in all detergent-solubilized AcrB samples and in the AcrB crystal. The identity of this ligand was further investigated using phospholipid analysis of purified AcrB and AcrB variant samples, and indicated the presence of phosphatidylethanolamine and possibly cardiolipin, both constituents of the Escherichia coli membrane. PMID- 21616056 TI - Crystal structure of the cell corpse engulfment protein CED-2 in Caenorhabditis elegans. AB - In the nematode Caenorhabditis elegans, the cell corpse engulfment proteins CED 2, CED-5, and CED-12 act in the same pathway to regulate the activation of the Rac small GTPase, CED-10, leading to the rearrangement of the actin cytoskeleton for engulfing apoptotic cells. Nevertheless, it is not well understood how these proteins act together. Here we report the crystal structures of the CED-2 protein as determined by X-ray crystallography. The full-length CED-2 protein and its truncated form containing the N-terminal SH2 domain and the first SH3 domain show similar three-dimensional structures. A CED-2 point mutation (F125G) disrupting its interaction with the PXXP motif of CED-5 did not affect its rescuing activity. However, CED-2 was found to interact with the N-terminal region of CED 5. Our findings suggest that CED-2 may regulate cell corpse engulfment by interacting with CED-5 through the N-terminal region rather than the PXXP motif. PMID- 21616057 TI - Structure of a novel class II phospholipase D: catalytic cleft is modified by a disulphide bridge. AB - Phospholipases D (PLDs) are principally responsible for the local and systemic effects of Loxosceles envenomation including dermonecrosis and hemolysis. Despite their clinical relevance in loxoscelism, to date, only the SMase I from Loxosceles laeta, a class I member, has been structurally characterized. The crystal structure of a class II member from Loxosceles intermedia venom has been determined at 1.7A resolution. Structural comparison to the class I member showed that the presence of an additional disulphide bridge which links the catalytic loop to the flexible loop significantly changes the volume and shape of the catalytic cleft. An examination of the crystal structures of PLD homologues in the presence of low molecular weight compounds at their active sites suggests the existence of a ligand-dependent rotamer conformation of the highly conserved residue Trp230 (equivalent to Trp192 in the glycerophosphodiester phosphodiesterase from Thermus thermophofilus, PDB code: 1VD6) indicating its role in substrate binding in both enzymes. Sequence and structural analyses suggest that the reduced sphingomyelinase activity observed in some class IIb PLDs is probably due to point mutations which lead to a different substrate preference. PMID- 21616058 TI - Dynamic changes in hair cell stereocilia and cochlear transduction after noise exposure. AB - The structures of cochlear transduction include stereocilia at the apical surface of hair cells and their connection to the tectorial membrane. The transduction site is one of the loci for noise-induced cochlear damage. Although stereocilia are susceptible to noise, it has been found that in the inner ears of avians, this fragile structure is largely self-repairing and is associated with recovery of hearing sensitivity after noise exposure, as observed in the difference between the temporal threshold shift (TTS) and the permanent threshold shift (PTS). In the mammalian cochleae, however, threshold shifts measured in the auditory brainstem responses (ABR) did not parallel the chronological changes in the stereocilia on hair cells. It is unclear how the morphological recovery of the stereocilia on the mammalian hair cells is correlated with the changes in cochlear transduction that can be assessed by measuring receptor potential. In the present study, guinea pigs were exposed to a broadband noise of 110 dB SPL for 2h. Auditory sensitivity was evaluated using ABR and cochlear transduction was assessed using cochlear microphonics (CM). Stereocilia morphology was quantified at different time points after the noise and compared with the control. The noise produced a TTS of 55.69 +/- 14.13 dB in frequency-averaged ABR thresholds. The threshold shift was reduced to 9.58 +/- 11.75 dB SPL 1 month later with virtually no loss of hair cells. Damage to the stereocilia immediately after noise exposure was found to be associated with depression of CM amplitude. Virtually no abnormal stereocilia were observed 1month after the noise in association with a fully recovered CM. PMID- 21616059 TI - Activity-dependent SUMOylation of the brain-specific scaffolding protein GISP. AB - G-protein coupled receptor interacting scaffold protein (GISP) is a multi-domain, brain-specific protein derived from the A-kinase anchoring protein (AKAP)-9 gene. Using yeast two-hybrid screens to identify GISP interacting proteins we isolated the SUMO conjugating enzyme Ubc9. GISP interacts with Ubc9 in vitro, in heterologous cells and in neurons. SUMOylation is a post-translational modification in which the small protein SUMO is covalently conjugated to target proteins, modulating their function. Consistent with its interaction with Ubc9, we show that GISP is SUMOylated by both SUMO-1 and SUMO-2 in both in vitro SUMOylation assays and in mammalian cells. Intriguingly, SUMOylation of GISP in neurons occurs in an activity-dependent manner in response to chemical LTP. These data suggest that GISP is a novel neuronal SUMO substrate whose SUMOylation status is modulated by neuronal activity. PMID- 21616060 TI - TLR4 activates NF-kappaB in human ovarian granulosa tumor cells. AB - Previous studies have demonstrated expression of Toll-like receptors (TLRs) in the surface epithelium of normal ovaries (OSE) and in epithelial ovarian tumors. Most notably, OSE-derived cancers express TLR4, which activates the nuclear factor-kappa B (NF-kappaB) signaling cascade as a mediator of inflammatory response. Currently, there is considerable interest in elucidating the role of TLR-mediated signaling in cancers. Nevertheless, the expression of TLRs in granulosa cell tumors (GCTs) of the ovary, and the extent to which GCT expression of TLRs may influence cell-signaling pathways and/or modulate the efficacy of chemotherapeutics, has yet to be determined. In the present study, human GCT lines (COV434 and KGN) were utilized to evaluate expression of functional TLR4. TLR4 is expressed in GCT cell lines and ligation of TLR4 with bacterial lipopolysaccharide (LPS) led to IkappaB degradation and activation of NF-kappaB. NF-kappaB activation was confirmed by nuclear localization of NF-kappaB p65 following treatment with LPS and the naturally occurring ligand, HSP60. Notably, immunoneutralization of TLR4 blocked nuclear localization, and inhibition of NF kappaB signaling attenuated LPS-induced TNFalpha plus increased doubling time in both cell lines. Contradictory to reports using human OSE cell lines, inhibition of NF-kappaB signaling failed to sensitize GCT lines to TRAIL or cisplatin. In summary, findings herein are the first to demonstrate a functional TLR-signaling pathway specifically in GCTs, and indicate that in contrast to OSE-derived cancers, inhibition of NF-kappaB does not sensitize GCTs to TRAIL or cisplatin. PMID- 21616061 TI - Soluble FGFR4 extracellular domain inhibits FGF19-induced activation of FGFR4 signaling and prevents nonalcoholic fatty liver disease. AB - Fibroblast growth factor receptor 4 (FGFR4) is a transmembrane tyrosine kinase receptor that plays a crucial role in the regulation of hepatic bile acid and lipid metabolism. FGFR4 underlies high-fat diet-induced hepatic steatosis, suggesting that inhibition of FGFR4 activation may be an effective way to prevent or treat nonalcoholic fatty liver disease (NAFLD). To determine whether neutralization of FGFR4 ligands by soluble FGFR4 extracellular domain (FGFR4-ECD) can inhibit the activation of FGFR4, we constructed FGFR4-ECD expression vector and showed that FGFR4-ECD was effectively expressed in cells and secreted into culture medium. FGFR4-ECD inhibited FGF19-induced activation of FGFR4 signaling and reduced steatosis of HepG2 induced by palmitic acid in vitro. Furthermore, in a tetracycline-induced fatty liver model, expression of FGFR4-ECD in mouse liver reduced the accumulation of hepatic lipids and partially restored the expression of peroxisome proliferator-activated receptor alpha (PPARalpha), which promotes the mitochondrial fatty acid beta-oxidation but is repressed by tetracycline. Taken together, these results demonstrate that FGFR4-ECD can block FGFR4 signaling and prevent hepatic steatosis, highlighting the potential value of inhibition of FGFR4 signaling as a method for therapeutic intervention against NAFLD. PMID- 21616062 TI - Inhibition of human Na(v)1.5 sodium channels by strychnine and its analogs. AB - Strychnine and brucine from the seeds of the plant Strychnos nux vomica have been shown to have interesting pharmacological effects on several neurotransmitter receptors. In this study, we have characterized the pharmacological properties of strychnine and its analogs on human Na(v)1.5 channels to assess their potential therapeutic advantage in certain arrhythmias. Among the eight alkaloids, only strychnine and icajine exhibited inhibition potency on the Na(v)1.5 channel with the half-maximum inhibition (IC(50)) values of 83.1MUM and 104.6MUM, respectively. Structure-function analysis indicated that the increased bulky methoxy groups on the phenyl ring or the negatively charged oxygen atom may account for this lack of inhibition on the Na(v)1.5 channel. Strychnine and icajine may bind to the channel by cation-pi interactions. The substitution with a large side chain on the phenyl ring or the increased molecular volume may alter the optimized position for the compound close to the binding sites of the channel. Strychnine and icajine bind to the Na(v)1.5 channel with a new mechanism that is different from TTX and local anesthetics. They bind to the outer vestibule of the channel pore with fast association and dissociation rates at resting state. Strychnine and icajine had little effect on steady-state fast inactivation but markedly shifted the slow inactivation of Na(v)1.5 currents toward more hyperpolarized potentials. The property of icajine influencing slow inactivated state of Na(v)1.5 channel would be potential therapeutic advantages in certain arrhythmias. PMID- 21616063 TI - piRNA, the new non-coding RNA, is aberrantly expressed in human cancer cells. AB - BACKGROUND: Piwi-interacting RNAs (piRNAs) are a novel class of non-coding single strand RNAs. They are involved in germline development, in silencing of selfish DNA elements, and in maintaining germline DNA integrity. The relationship between piRNAs and carcinogenesis has not been shown yet. METHODS: The relationship between piRNAs and carcinogenesis was identified by microarray screening and real time quantitative reverse transcription-polymerase chain reaction technology. The piR-651 inhibitor was transfected into gastric cancer cells to assess its influence on cell growth. Cell cycle analysis was used to reveal the cellular mechanisms of piR-651 in the genesis of gastric cancer. RESULTS: piR-651 expression was upregulated in gastric cancer tissues compared with paired non cancerous tissues. The levels of piR-651 were associated with TNM stage (P=0.032). The expression of piR-651 in gastric, colon, lung, and breast cancer tissues was higher than that in paired non-cancerous tissues. The upregulated expression of piR-651 was confirmed in several cancer cell lines including gastric, lung, mesothelium, breast, liver, and cervical cancer cell lines. The growth of gastric cancer cells was inhibited by a piR-651 inhibitor and arrested at the G(2)/M phase. CONCLUSION: piR-651 might be involved in the development of gastric cancer and other cancers, and is a potential marker for cancer diagnosis. PMID- 21616064 TI - Guasha-induced hepatoprotection in chronic active hepatitis B: a case study. AB - BACKGROUND: Heme oxygenase-1 (HO-1) has demonstrated hepatoprotective effect in animal hepatitis models. HO-1 was also reported to be upregulated with Guasha, an ancient therapeutic technique which applies instrument assisted press-stroking to treat many disorders. METHODS: We report a case on the changes of liver function, plasma HO-1 and T-helper (Th) cytokine balance in a chronic active hepatitis B carrier before and after Guasha. The patient presented with increased activities of liver enzymes (ALT and AST), indicating inflammatory damage in liver before Guasha. RESULTS: Forty-eight hours after receiving Guasha, the patient showed changes in a number of serum markers: a decline of liver enzymes (ALT and AST) indicating reduced chronic inflammation, an elevated plasma HO-1, and a modulation of T-helper (Th)1/Th2 balance. CONCLUSIONS: Guasha was shown to transiently reduce the inflammatory markers of liver injury in human, together with an enhancement of HO-1 which might be responsible for the hepatoprotective action. PMID- 21616065 TI - Actions of two GABAA receptor benzodiazepine-site ligands that are mediated via non-gamma2-dependent modulation. AB - The potent sedative-hypnotic zolpidem and the convulsant methyl-6,7-dimethoxy-4 ethyl-beta-carboline-3-carboxylate (DMCM) act primarily by binding to the benzodiazepine site of the main inhibitory neurotransmitter receptor, the pentameric gamma-aminobutyric acid type A receptor (GABA(A)). This binding depends critically on the wild-type F77 residue of the GABA(A) receptor gamma2 subunit. Mice with gamma2 subunit F77I point mutation (gamma2I77 mouse line) lose the high-affinity nanomolar binding of these ligands as well as their most robust behavioral actions at low doses. Interestingly, the gamma2I77 mice offer a tool to study the actions of these substances mediated via other possible binding sites of the GABA(A) receptor. In ligand autoradiographic experiments, we discovered in gamma2I77 mouse brain sections a significant amount of residual non gamma2 subunit-dependent benzodiazepine site binding enriched to the striatum and septum. Zolpidem only weakly affected this residual binding at micromolar concentrations, and only a high zolpidem dose (>= 40 mg/kg) caused sedation and deficits in motor coordination in gamma2I77 mice. DMCM had an agonistic action through a secondary, low-affinity non-benzodiazepine binding site of the GABA(A) receptor in the forebrain of gamma2I77 mice, and this drug also fully displaced the residual benzodiazepine-site labeling. In behavioral tests, a high dose (20mg/kg) of DMCM was sedative and modulated fear learning. DMCM, but not zolpidem, acted as an agonist in recombinant GABA(A) alpha1/6beta3 receptors studied using ligand binding and electrophysiological assays. Our results highlight the less well-known actions of high doses of DMCM and zolpidem that are not mediated via the gamma2 subunit-containing benzodiazepine site of the GABA(A) receptor. PMID- 21616066 TI - Protective effects of saffron extract and its active constituent crocin against oxidative stress and spatial learning and memory deficits induced by chronic stress in rats. AB - Although it is well established that chronic stress impairs spatial learning and memory, few studies have investigated possible ways to prevent its deleterious effects. Here, we investigated the effects of Crocus sativus L., commonly known as saffron, and its active constituent crocin on learning and memory loss and the induction of oxidative stress in the hippocampus by chronic stress. Rats were injected with saffron extract, crocin or vehicle over a period of 21 days while being exposed to chronic restraint stress (6 h/day). After this, they were trained and tested on a water-maze spatial memory task. They performed four trials per day for 5 consecutive days, and this was followed by a probe trial two days later. At the end of the behavioral testing, several parameters of oxidative stress in the hippocampus were measured. Treatment with saffron extract or crocin blocked the ability of chronic stress to impair spatial learning and memory retention. Relative to controls that received vehicle, stressed animals that received saffron extract or crocin had significantly higher levels of lipid peroxidation products, significantly higher activities of antioxidant enzymes including glutathione peroxidase, glutathione reductase and superoxide dismutase and significantly lower total antioxidant reactivity capacity. Finally, crocin significantly decreased plasma levels of corticosterone, as measured after the end of stress. These observations indicate that saffron and its active constituent crocin can prevent the impairment of learning and memory as well as the oxidative stress damage to the hippocampus induced by chronic stress. Thus, using these substances may be useful in pharmacological alleviation of cognitive deficits. PMID- 21616067 TI - Schistosoma mansoni: structural and biochemical characterization of two distinct Venus Kinase Receptors. AB - Venus Kinase Receptors (VKRs) are atypical transmembrane proteins composed of an extracellular Venus FlyTrap module linked through a single helix to a tyrosine kinase domain similar to that of insulin receptors. This structure was first described in Schistosoma mansoni, then in a selected range of invertebrates, including many insects. The preferential expression of VKRs in larvae and gonads suggested their role in development and reproduction. While a single vkr gene was consistently found in all genomes, we identified two distinct vkr genes in S. mansoni. Our data indicated that Smvkr1 and Smvkr2 are very similar in structure and likely originated from gene duplication. Both genes are expressed in all the parasite stages and encode homologous proteins with a conserved VKR structure. Recombinant SmVKR1 and SmVKR2 exhibit tyrosine kinase activities dependent on the binding of distinct small ligand molecules. SmVKR1 and SmVKR2 could represent paralogs with different functions in the parasite. PMID- 21616068 TI - Schistosoma mansoni egg glycoproteins and C-type lectins of host immune cells: molecular partners that shape immune responses. AB - Schistosome eggs and egg-derived molecules are potent immunomodulatory agents. There is increasing evidence that the interplay between egg glycoproteins and host C-type lectins plays an important role in shaping immune responses during schistosomiasis. As most experiments in this field so far have been performed using complex protein/glycoprotein mixtures or synthetic model glycoconjugates, it is still largely unclear which individual moieties of schistosome eggs are immunologically active. In this review we will discuss molecular aspects of Schistosoma mansoni egg glycoproteins, their interactions with C-type lectins, and the relevance to schistosome egg immunobiology. PMID- 21616069 TI - Trypanosoma evansi: Ca(2+) ATPase activity and lipid peroxidation in skeletal muscle from rats experimentally infected. AB - The aim of this study was to evaluate Ca(2+) ATPase activity and the lipid peroxidation in muscles from rats experimentally infected by Trypanosoma evansi and its roles in the muscle pathogenesis in trypanosomosis. Thirty-six rats were divided in two groups. Group A was infected with an isolate from T. evansi and group B was used as a negative control. Group A was divided into three subgroups (A1, A2 and A3), three animals each group, as well as group B (B1, B2 and B3). The collection of samples were performed at days 5 (A1 and B1), 15 (A2 and B2) and 30 (A3 and B3) post-infection (PI) with the purpose of comparison between healthy and infected rats in the course of the disease. The Ca(2+) ATPase enzyme activity was determined in skeletal muscle samples. Muscle tissue lipid peroxidation was determined by TBARS levels, and histopathologically it was investigated a possible damage to the muscle tissue of rats infected with T. evansi. It was observed a significant decrease of Ca(2+) ATPase activity in infected rats compared to not-infected. This enzymatic inhibition was observed at days 5, 15 and 30 PI. A significant increase was observed for TBARS levels in the muscles of infected rats at days 5, 15 and 30 PI. It was not identified any histological alterations for gastrocnemius in rats infected by T. evansi at days 5 and 15 PI. Nevertheless, at day 30 PI it was verified inflammatory infiltrate with mononuclear cells between muscle fibers in three infected rats (50%). T. evansi infections in rats showed a negative correlation between Ca(2+) ATPase and TBARS levels. Based on these results we suggest that the leg weakness and muscle injuries common in infected animals with T. evansi may be related to a reduced activity of Ca(2+) ATPase and oxidative stress. PMID- 21616070 TI - Toxoplasma gondii aspartic protease 1 is not essential in tachyzoites. AB - Aspartic proteases are important virulence factors for pathogens and are recognized as attractive drug targets. Seven aspartic proteases (ASPs) have been identified in Toxoplasma gondii genome. Bioinformatics and phylogenetic analyses regroup them into five monophyletic groups. Among them, TgASP1, a coccidian specific aspartic protease related to the food vacuole plasmepsins, is associated with the secretory pathway in non-dividing cells and relocalizes in close proximity to the nascent inner membrane complex (IMC) of daughter cells during replication. Despite a potential role for TgASP1 in IMC formation, the generation of a conventional knockout of the TgASP1 gene revealed that this protease is not required for T. gondii tachyzoite survival or for proper IMC biogenesis. PMID- 21616071 TI - Leishmania aethiopica: development of specific and sensitive PCR diagnostic test. AB - PCR has proved useful for rapid diagnosis and typing of Leishmania. Lack of specificity to discriminate between species and/or sensitivity to detect from clinical samples has always been an issue. Previously developed primers either require PCR-RFLP analysis for Leishmania aethiopica discrimination or lack sensitivity to detect L. aethiopica from clinical samples. Here we report the development and validation of L. aethiopica specific PCR primers (V5F/V10R) based on cysteine protease B (cpb), a multicopy and polymorphic gene of Leishmania. V5F/V10R primers differentiate L. aethiopica from Leishmania tropica, Leishmania major, Leishmania donovani and Leishmania infantum by direct PCR. In addition, they are sensitive enough to detect L. aethiopica from biopsy samples. The primers can be very useful for epidemiological studies, species typing and diagnosis of L. aethiopica directly from clinical samples. Implementation of these primers in routine L. aethiopica diagnosis can improve detection rate, save time, money and labor required for culturing Leishmania. PMID- 21616072 TI - Plasmodium falciparum: DNA sequence specificity of cisplatin and cisplatin analogues. AB - In this paper, we provided evidence that cisplatin is able to form adducts with cellular DNA in Plasmodium falciparum. The DNA sequence specificity of cisplatin adduct formation was determined in trophozoite-enriched P. falciparum cells and this paper represents the first occasion that the sequence specificity of cisplatin DNA damage has been observed in malaria cells. Utilising a sub telomeric, 692 bp repeat sequence in the P. falciparum genome, we were able to investigate the DNA adducts formed by cisplatin and five analogues. A run of eight consecutive guanines was the most prominent site of DNA damage in the malarial cells. This study suggests that the mechanism of P. falciparum cell death caused by cisplatin involves damage to DNA and hence inhibition of DNA replication and cell division. PMID- 21616073 TI - The type III secretion system is involved in Escherichia coli K1 interactions with Acanthamoeba. AB - The type III secretion system among Gram-negative bacteria is known to deliver effectors into host cell to interfere with host cellular processes. The type III secretion system in Yersina, Pseudomonas and Enterohemorrhagic Escherichia coli have been well documented to be involved in the bacterial pathogenicity. The existence of type III secretion system has been demonstrated in neuropathogenic E. coli K1 strains. Here, it is observed that the deletion mutant of type III secretion system in E. coli strain EC10 exhibited defects in the invasion and intracellular survival in Acanthamoeba castellanii (a keratitis isolate) compared to its parent strain. Next, it was determined whether type III secretion system plays a role in E. coli K1 survival inside Acanthamoeba during the encystment process. Using encystment assays, our findings revealed that the type III secretion system-deletion mutant exhibited significantly reduced survival inside Acanthamoeba cysts compared with its parent strain, EC10 (P<0.01). This is the first demonstration that the type III secretion system plays an important role in E. coli interactions with Acanthamoeba. A complete understanding of how amoebae harbor bacterial pathogens will help design strategies against E. coli transmission to the susceptible hosts. PMID- 21616074 TI - Lipin-1gamma isoform is a novel lipid droplet-associated protein highly expressed in the brain. AB - Lipin-1 proteins are phosphatidic acid phosphatases (PAPs) catalyzing the conversion from phosphatidic acid (PA) to diacylglycerol (DG). Two alternative splicing isoforms, lipin-1alpha and -1beta, are localized at different subcellular compartments. A third splicing isoform, lipin-1gamma was recently cloned and its subcellular localization is unknown. Here, we demonstrate that lipin-1gamma is localized to lipid droplets (LDs), an association mediated by a hydrophobic, lipin-1gamma-specific domain. Additional expression of lipin-1gamma altered LD morphology without affecting the triacylglycerol (TG) level. In human tissues, lipin-1gamma is the main lipin-1 isoform expressed in normal human brain, suggesting a specialized role in regulating brain lipid metabolism. PMID- 21616076 TI - Quantitative short-day photoperiodic response in larval development and its adaptive significance in an adult-overwintering cerambycid beetle, Phytoecia rufiventris. AB - The chrysanthemum longicorn beetle, Phytoecia rufiventris, overwinters in the adult stage and reproduces in spring. Larvae of this beetle develop during summer inside a host stem or root. In the present study, photoperiodic control of larval development and its adaptive significance were examined in this beetle using an artificial diet. Larvae showed a short-day photoperiodic response at 25 degrees C with a critical day length of around 14 h; larvae reared under short-day conditions pupated, whereas those reared under long-day conditions entered summer diapause with some supernumerary molts and did not pupate. A similar response was found at 30 degrees C, but with a shorter critical day length. Below the critical day length, a shorter day length corresponded to a shorter larval period. Larvae transferred from long-day conditions to various photoperiods showed a similar quantitative response. Field rearing of larvae starting at various times of year showed that pupation occurs within a relatively short period in early autumn. Field rearing of pupae and adults at various times indicated that only pupation in early autumn results in a high survival rate until winter. Earlier or later pupation led to a low survival rate due to death before overwintering in the adult and pupal stages, respectively. Thus, in P. rufiventris, timing of pupation regulated by the quantitative short-day photoperiodic response is vital for survival. Relatively lower developmental threshold in the pupal stage supports this hypothesis. PMID- 21616075 TI - Neuronal plasticity in the cingulate cortex of rats following esophageal acid exposure in early life. AB - BACKGROUND & AIMS: The cingulate cortex has been reported to be involved in processing pain of esophageal origin. However, little is known about molecular changes and cortical activation that arise from early-life esophageal acid reflux. Excitatory neurotransmission via activation of the N-methyl-d-aspartate (NMDA) receptor and its interaction with postsynaptic density protein 95 (PSD-95) at the synapse appear to mediate neuronal development and plasticity. We investigated the effect of early-life esophageal acid exposure on NMDA receptor subunits and PSD-95 expression in the developing cingulate cortex. METHODS: We assessed NMDA receptor subunits and PSD-95 protein expression in rostral cingulate cortex (rCC) tissues of rats exposed to esophageal acid or saline (control), either during postnatal day (P) 7 to 14 and/or acutely at adult stage (P60) using immunoblot and immunoprecipitation analyses. RESULTS: Compared with controls, acid exposure from P7 to P14 significantly increased expression of NR1, NR2A, and PSD-95, measured 6 weeks after exposure. However, acute exposure at P60 caused a transient increase in expression of NMDA receptor subunits. These molecular changes were more robust in animals exposed to acid neonatally and rechallenged, acutely, at P60. Esophageal acid exposure induced calcium calmodulin kinase II-mediated phosphorylation of the subunit NR2B at Ser1303. CONCLUSIONS: Esophageal acid exposure during early stages of life has long-term effects as a result of phosphorylation of the NMDA receptor and overexpression in the rCC. This molecular alteration in the rCC might mediate sensitization of patients with acid-induced esophageal disorders. PMID- 21616077 TI - Detection of single and mixed covert baculovirus infections in eastern spruce budworm, Choristoneura fumiferana populations. AB - We surveyed for covert baculovirus infections in the eastern spruce budworm, Choristoneura fumiferana (Clemens) and compared the prevalence of virus detected in a laboratory and a field population. DNA was extracted from budworm adults and then PCR with degenerate primers was used to identify individuals carrying baculovirus DNA. Multiplex PCR was then applied to the positive samples to distinguish between the multiple baculovirus types that could potentially be found in C. fumiferana populations. Covert infections were found in both the laboratory and the field population of C. fumiferana, although the frequency of infection and the composition of viruses found were very different. Overall 28% of insects from the laboratory population were positive for baculovirus DNA. Individual adults supported both single and mixed covert infections with CfMNPV plus CfDEFNPV, CfDEFNPV plus a GV and mixtures of all three viruses together. However, the majority of insects supported single virus infections, and surprisingly this virus was CfDEFNPV, a virus that is reported not to have per os activity in C. fumiferana larvae. Insects from field populations showed a very different pattern; 70.5% of individuals were baculovirus positive and all of these were positive for CfDEFNPV only. PMID- 21616078 TI - MCP-1-induced protein attenuates endotoxin-induced myocardial dysfunction by suppressing cardiac NF-kB activation via inhibition of IkB kinase activation. AB - Myocardial contractile dysfunction is a major consequence of septic shock, which is mainly mediated by nuclear factor-kappa B (NF-kB)-dependent production of inflammatory mediators in the heart. A novel zinc-finger protein, MCP-1-induced protein (MCPIP), is thought to have NF-kB inhibitory activity in certain cell cultures, but its pathophysiological consequence in vivo remains undefined. This study aims to clarify whether the anti-inflammatory potency of MCPIP contribute to amelioration of septic myocardial inflammation and dysfunction in vivo. Transgenic mice (TG) with cardiac-specific expression of MCPIP and their littermate wild-type (WT) controls were challenged with Escherichia coli LPS (10mg/kg ip) and myocardial function was assessed 18 h later using echocardiography. LPS administration markedly deteriorated myocardial contractile function evidenced by reduction of the percentage of left ventricular fractional shortening, which was significantly attenuated by myocardial expression of MCPIP. MCPIP TG mice exhibited a markedly reduced myocardial inflammatory cytokines, less of iNOS expression and peroxynitrite formation, decreased caspase-3/7 activities and apoptotic cell death compared with LPS-treated WT mice. Activation of cardiac NF-kB observed in LPS-challenged WT mice was suppressed by the presence of MCPIP, as evidenced by decreased phosphorylation of IkB kinase (IKKalpha/beta), reduced degradation of the cytosolic IkBalpha, and decreased nuclear translocation of NF-kB p65 subunit and its target DNA-binding activity. These results suggest that MCPIP has therapeutic values to protect heart from inflammatory pathologies, possibly through inhibition of IkB kinase complex, leading to blockade of NF-kB activation, and subsequently, attenuation of the proinflammatory state and nitrosative stress in the myocardium. PMID- 21616079 TI - Cardiac myosin activation part 1: from concept to clinic. AB - Decreased cardiac contractility is a central feature of systolic heart failure and yet we have no effective drugs to improve cardiac contractility. Existing drugs that increase cardiac contractility do so indirectly through signaling cascades and their use is limited by their mechanism-related adverse effects. Direct activation of the cardiac sarcomere to increase cardiac contractility may provide a means to avoid these limitations. Using a reconstituted version of the cardiac sarcomere, we screened a small molecule library and identified several chemical classes that directly activate cardiac myosin. One compound class has been optimized extensively using an iterative process; omecamtiv mecarbil, a small-molecule, selective, cardiac myosin activator is the most advanced exemplar of this novel mechanistic class. It accelerates the transition of myosin into the force-generating state without affecting cardiac myocyte calcium homeostasis. In animal models, omecamtiv mecarbil increases cardiac function by increasing the duration of ejection without changing the rates of contraction. Initial clinical studies have demonstrated the translation of this mechanism into humans, and further clinical studies of its use in acute and chronic heart failure are planned. Cardiac myosin activation may provide a new therapeutic approach for systolic heart failure. This article is part of a special issue entitled "Key Signaling Molecules in Hypertrophy and Heart Failure." PMID- 21616081 TI - Monte Carlo study of the formation and conformational properties of dimers of Abeta42 variants. AB - Small soluble oligomers, and dimers in particular, of the amyloid beta-peptide (Abeta) are believed to play an important pathological role in Alzheimer's disease. Here, we investigate the spontaneous dimerization of Abeta42, with 42 residues, by implicit solvent all-atom Monte Carlo simulations, for the wild-type peptide and the mutants F20E, E22G and E22G/I31E. The observed dimers of these variants share many overall conformational characteristics but differ in several aspects at a detailed level. In all four cases, the most common type of secondary structure is intramolecular antiparallel beta-sheets. Parallel, in-register beta sheet structure, as in models for Abeta fibrils, is rare. The primary force driving the formation of dimers is hydrophobic attraction. The conformational differences that we do see involve turns centered in the 20-30 region. The probability of finding turns centered in the 25-30 region, where there is a loop in Abeta fibrils, is found to increase upon dimerization and to correlate with experimentally measured rates of fibril formation for the different Abeta42 variants. Our findings hint at reorganization of this part of the molecule as a potentially critical step in Abeta aggregation. PMID- 21616080 TI - A two-site mechanism for the inhibition of IAPP amyloidogenesis by zinc. AB - Human islet amyloid polypeptide (hIAPP) is a highly amyloidogenic protein co secreted with insulin in response to glucose levels. The formation of hIAPP amyloid plaques near islet cells has been linked to the death of insulin secreting beta-cells in humans and the progression of type II diabetes. Since both healthy individuals and those with type II diabetes produce and secrete hIAPP, it is reasonable to look for factors involved in storing hIAPP and preventing amyloidosis. We have previously shown that zinc inhibits the formation of insoluble amyloid plaques of hIAPP; however, there remains significant ambiguity in the underlying mechanisms. In this study, we show that zinc binds unaggregated hIAPP at micromolar concentrations similar to those found in the extracellular environment. By contrast, the fibrillar amyloid form of hIAPP has low affinity for zinc. The binding stoichiometry obtained from isothermal titration calorimetry experiments indicates that zinc favors the formation of hIAPP hexamers. High-resolution NMR structures of hIAPP bound to zinc reveal changes in the electron environment along residues that would be located along one face of the amphipathic hIAPP alpha-helix proposed as an intermediate for amyloid formation. Results from electrospray ionization mass spectroscopy investigations showed that a single zinc atom is predominantly bound to hIAPP and revealed that zinc inhibits the formation of the dimer. At higher concentrations of zinc, a second zinc atom binds to hIAPP, suggesting the presence of a low affinity secondary binding site. Combined, these results suggest that zinc promotes the formation of oligomers while creating an energetic barrier for the formation of amyloid fibers. PMID- 21616083 TI - Percent of highly immunogenic amino acid residues forming B-cell epitopes is higher in homologous proteins encoded by GC-rich genes. AB - We analyzed the dependence of the percent of highly immunogenic amino acid residues included in B-cell epitopes of homologous proteins on the GC-content (G+C) of genes coding for them in twenty-seven lineages of proteins (and subsequent genes), which belong to seven Varicello and five Simplex viruses. We found out that proteins encoded by genes of a high GC-content usually contain more targets for humoral immune response than their homologs encoded by GC-poor genes. This tendency is characteristic not only to the lineages of glycoproteins, which are the main targets for humoral immune response against Simplex and Varicello viruses, but also to the lineages of capsid proteins and even "housekeeping" enzymes. The percent of amino acids included in linear B-cell epitopes has been predicted for 324 proteins by BepiPred algorithm (www.cbs.dtu.dk/services/BepiPred), the percent of highly immunogenic amino acids included in discontinuous B-cell epitopes and the percent of exposed amino acid residues have been predicted by Epitopia algorithm (http://epitopia.tau.ac.il/). Immunological consequences of the directional mutational GC-pressure are mostly due to the decrease in the total usage of highly hydrophobic amino acids and due to the increase in proline and glycine levels of usage in proteins. The weaker the negative selection on amino acid substitutions caused by symmetric mutational pressure, the higher the slope of direct dependence of the percent of highly immunogenic amino acids included in B-cell epitopes on G+C. PMID- 21616082 TI - Characterization of the structure and function of Klebsiella pneumoniae allantoin racemase. AB - The oxidative catabolism of uric acid produces 5-hydroxyisourate (HIU), which is further degraded to (S)-allantoin by two enzymes, HIU hydrolase and 2-oxo-4 hydroxy-4-carboxy-5-ureidoimidazoline decarboxylase. The intermediates of the latter two reactions, HIU and 2-oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline, are unstable in solution and decay nonstereospecifically to allantoin. In addition, nonenzymatic racemization of allantoin has been shown to occur at physiological pH. Since the further breakdown of allantoin is catalyzed by allantoinase, an enzyme that is specific for (S)-allantoin, an allantoin racemase is necessary for complete and efficient catabolism of uric acid. In this work, we characterize the structure and activity of allantoin racemase from Klebsiella pneumoniae (KpHpxA). In addition to an unliganded structure solved using selenomethionyl single wavelength anomalous dispersion, structures of C79S/C184S KpHpxA in complex with allantoin and with 5-acetylhydantoin are presented. These structures reveal several important features of the active site including an oxyanion hole and a polar binding pocket that interacts with the ureido tail of allantoin and serves to control the orientation of the hydantoin ring. The ability of KpHpxA to interconvert the (R)- and (S)-enantiomers of allantoin is demonstrated, and analysis of the steady-state kinetics of KpHpxA yielded a k(cat)/K(m) of 6.0 * 10(5) M(-1) s(-1). Mutation of either of the active-site cysteines, Cys79 or Cys184, to serine inactivates this enzyme. The data presented provide new insights into the activity and substrate specificity of this enzyme and enable us to propose a mechanism for catalysis that is consistent with the two-base mechanism observed in other members of the aspartate/glutamate family. PMID- 21616084 TI - Focal adhesion kinase regulation of neovascularization. AB - In this review, we discuss the role of focal adhesion kinase (FAK), an intracellular tyrosine kinase, in endothelial cells in relation to neovascularization. Genetic and in vitro studies have identified critical factors, receptor systems, and their intracellular signaling components that regulate the neovasculogenic phenotypes of endothelial cells. Among these factors, FAK appears to regulate several aspects of endothelial cellular behavior, including migration, survival, cytoskeletal organization, as well as cell proliferation. Upon adhesion of endothelial cells to extracellular matrix (ECM) ligands, integrins cluster on the plane of plasma-membrane, while cytoplasmic domains of integrins interact with cytoskeletal proteins and signaling molecules including FAK. However, FAK not only serves as a critical component of integrin signaling, but is also a downstream element of the VEGF/VEGF-receptor and other ligand-receptor systems that regulate neovascularization. A complete understanding of FAK-mediated neovascularization, therefore, should address the molecular and cellular mechanisms that regulate the biology of FAK. Continued research on FAK may, therefore, yield novel therapies to improve treatment modalities for the pathological neovascularization associated with diseases. PMID- 21616085 TI - MicroRNA expression in the livers of inbred mice. AB - MicroRNAs are short, non-coding RNA sequences that regulate genes at the post transcriptional level and have been shown to be important in development, tissue differentiation, and disease. Limited attention has been given to the natural variation in miRNA expression across genetically diverse populations even though it is well established that genetic polymorphisms can have a profound effect on mRNA levels. Expression level of 577 miRNAs in the livers of 70 strains of inbred mice was assessed, and we found that miRNA expression is highly stable across different strains. Globally, the expression of miRNA target transcripts does not correlate with miRNA expression, primarily due to the low variance of miRNA but high variance of mRNA expression across strains. Our results show that there is little genetic effect on the baseline miRNA levels in murine liver. The stability of mouse liver miRNA expression in a genetically diverse population suggests that treatment-induced disruptions in liver miRNA expression, a phenomenon established for a large number of toxicants, may indicate an important mechanism for the disturbance of normal liver function, and may prove to be a useful genetic background-independent biomarker of toxicant effect. PMID- 21616087 TI - Equine behaviour and heart rate in temperament tests with or without rider or handler. AB - The aim of the present study was to compare horses' heart rate (HR), heart rate variability (RMSSD, pNN50) and behaviour in the same temperament test when being ridden, led, and released free. Behavioural measurements included scores and linear measurements for reactivity (R), activity (A), time to calm down (T) and emotionality (E), recorded during the approach (1) and/or during confrontation with the stimulus (2). Sixty-five horses were each confronted 3 times (1 ridden, 1 led, 1 free running in balanced order) with 3 novel and/or sudden stimuli. Mixed model analysis indicated that leading resulted in the lowest (P<0.05 throughout) reactions as measured by A1, A2, E1, E2, R2, and pNN50 while riding produced the strongest (A1, T2, HR, RMSSD, pNN50) or medium (E1, E2, R2) reactions. Free running resulted either in the strongest (A2, E1, E2, R2) or in the lowest (A1, T2, HR, RMSSD, pNN50) reactions. The repeatability across tests for HR (0.57), but not for RMSSD (0.23) or pNN50 (0.25) was higher than for any behavioural measurement: the latter ranged from values below 0.10 (A1, A2, T2) to values between 0.30 and 0.45 (E1, E2, R2). Overall, the results show that a rider or handler influences, but not completely masks, the horses' intrinsic behaviour in a temperament test, and this influence appeared to be stronger on behavioural variables and heart rate variability than on the horses' heart rates. Taking both practical considerations and repeatabilities into account, reactivity appears to be the most valuable parameter. Emotionality and heart rate can also yield valid results reflecting additional dimensions of temperament although their practical relevance may be less obvious. If a combination of observed variables is chosen with care, a valid assessment of a horse's temperament may be possible in all types of tests. However, in practice, tests that resemble the practical circumstances most closely, i.e. testing riding horses under a rider, should be chosen. PMID- 21616086 TI - Functional dissociations between four basic arithmetic operations in the human posterior parietal cortex: a cytoarchitectonic mapping study. AB - Although lesion studies over the past several decades have focused on functional dissociations in posterior parietal cortex (PPC) during arithmetic, no consistent view has emerged of its differential involvement in addition, subtraction, multiplication, and division. To circumvent problems with poor anatomical localization, we examined functional overlap and dissociations in cytoarchitectonically defined subdivisions of the intraparietal sulcus (IPS), superior parietal lobule (SPL) and angular gyrus (AG), across these four operations. Compared to a number identification control task, all operations except addition, showed a consistent profile of left posterior IPS activation and deactivation in the right posterior AG. Multiplication and subtraction differed significantly in right, but not left, IPS and AG activity, challenging the view that the left AG differentially subserves retrieval during multiplication. Although addition and multiplication both rely on retrieval, multiplication evoked significantly greater activation in right posterior IPS, as well as the prefrontal cortex, lingual and fusiform gyri, demonstrating that addition and multiplication engage different brain processes. Comparison of PPC responses to the two pairs of inverse operations: division versus multiplication and subtraction versus addition revealed greater activation of left lateral SPL during division, suggesting that processing inverse relations is operation specific. Our findings demonstrate that individual IPS, SPL and AG subdivisions are differentially modulated by the four arithmetic operations and they point to significant functional heterogeneity and individual differences in activation and deactivation within the PPC. Critically, these effects are related to retrieval, calculation and inversion, the three key cognitive processes that are differentially engaged by arithmetic operations. Our findings point to distribute representation of these processes in the human PPC and also help explain why lesion and previous imaging studies have yielded inconsistent findings. PMID- 21616088 TI - Metabolism of aflatoxin B1 in turkey liver microsomes: the relative roles of cytochromes P450 1A5 and 3A37. AB - The extreme sensitivity of turkeys to aflatoxin B(1) (AFB(1)) is associated with efficient epoxidation by hepatic cytochromes P450 (P450) 1A5 and 3A37 to exo aflatoxin B(1)-8,9-epoxide (exo-AFBO). The combined presence of 1A5 and 3A37, which obey different kinetic models, both of which metabolize AFB(1) to the exo AFBO and to detoxification products aflatoxin M(1) (AFM(1)) and aflatoxin Q(1) (AFQ(1)), respectively, complicates the kinetic analysis of AFB(1) in turkey liver microsomes (TLMs). Antisera directed against 1A5 and 3A37, thereby individually removing the catalytic contribution of these enzymes, were used to identify the P450 responsible for epoxidating AFB(1) in TLMs. In control TLMs, AFB(1) was converted to exo-AFBO in addition to AFM(1) and AFQ(1) confirming the presence of functional 1A5 and 3A37. Pretreatment with anti-1A5 inhibited exo AFBO formation, especially at low, submicromolar (~0.1MUM), while anti-3A37, resulted in inhibition of exo-AFBO formation, but at higher (>50MUM) AFB(1) concentrations. Metabolism in immunoinhibited TLMs resembled that of individual enzymes: 1A5 produced exo-AFBO and AFM(1), conforming to Michaelis-Menten, while 3A37 produced exo-AFBO and AFQ(1) following the kinetic Hill equation. At 0.1MUM AFB(1), close to concentrations in livers of exposed animals, 1A5 contributed to 98% of the total exo-AFBO formation. At this concentration, 1A5 accounted for a higher activation:detoxification (50:1, exo-AFBO: AFM(1)) compared to 3A37 (0.15: 1, exo-AFBO: AFQ(1)), suggesting that 1A5 is high, while 3A4 is the low affinity enzyme in turkey liver. The data support the conclusion that P450 1A5 is the dominant enzyme responsible for AFB(1) bioactivation and metabolism at environmentally-relevant AFB(1) concentrations in turkey liver. PMID- 21616090 TI - 5-Methoxyflavanone induces cell cycle arrest at the G2/M phase, apoptosis and autophagy in HCT116 human colon cancer cells. AB - Natural flavonoids have diverse pharmacological activities, including anti oxidative, anti-inflammatory, and anti-cancer activities. In this study, we investigated the molecular mechanism underlying the action of 5-methoxyflavanone (5-MF) which has a strong bioavailability and metabolic stability. Our results show that 5-MF inhibited the growth and clonogenicity of HCT116 human colon cancer cells, and that it activated DNA damage responses, as revealed by the accumulation of p53 and the phosphorylation of DNA damage-sensitive proteins, including ataxia-telangiectasia mutated (ATM) at Ser1981, checkpoint kinase 2 (Chk2) at Thr68, and histone H2AX at Ser139. 5-MF-induced DNA damage was confirmed in a comet tail assay. We also found that 5-MF increased the cleavage of caspase-2 and -7, leading to the induction of apoptosis. Pretreatment with the ATM inhibitor KU55933 enhanced 5-MF-induced gamma-H2AX formation and caspase-7 cleavage. HCT116 cells lacking p53 (p53(-/-)) or p21 (p21(-/-)) exhibited increased sensitivity to 5-MF compared to wild-type cells. 5-MF further induced autophagy via an ERK signaling pathway. Blockage of autophagy with the MEK inhibitor U0126 potentiated 5-MF-induced gamma-H2AX formation and caspase-2 activation. These results suggest that a caspase-2 cascade mediates 5-MF-induced anti-tumor activity, while an ATM/Chk2/p53/p21 checkpoint pathway and ERK mediated autophagy act as a survival program to block caspase-2-mediated apoptosis induced by 5-MF. PMID- 21616091 TI - Evaluation of a ricin vaccine candidate (RVEc) for human toxicity using an in vitro vascular leak assay. AB - To protect against ricin intoxication, a genetically derived ricin A chain vaccine candidate (RVEc) was developed lacking the toxic N-glycosidase activity (Olson et al., 2004). The vaccine protects animals against an aerosolized ricin holotoxin (RT) challenge (Carra et al., 2007). In the current study, the RVEc vaccine was evaluated for its interaction and effect on human endothelial cells. RVEc was tested in an in vitro cellular-based bioassay, consisting of primary human endothelial cells cultured on collagen-coated inserts, to which concentrations of the vaccine candidate (0.6, 2, 2.5 or 9 MUM) were added. RVEc showed no signs of adverse activity on the cells (e.g., cytotoxicty activity) as measured by changes in trans-endothelial electrical resistance (TEER). In contrast, ricin toxin (RT) cytotoxicity was observed at all concentrations tested. Under light microscopy, no cytotoxicity was visible at 24h with 0.6 or 9 MUM of RVEc. However, cytotoxicity was observed for RT and to a lesser degree for RTA. Flow cytometric analysis showed binding of RT, slight binding of RTA, and no binding of the RVEc vaccine to endothelial cells. The presence of RTB as a contaminant contributing to the cytotoxicity in the RTA preparation was ruled out by a RTB-specific ELISA. In addition, RTA at 9 MUM produced a cytotoxic activity that could not be explained exclusively by the presence of azide in the RTA buffer. In the current study, the model demonstrated no discernable adverse events of the RVEc vaccine on human endothelial cells, when compared to the toxicity caused by holotoxin or native RTA preparations. PMID- 21616089 TI - The aryl hydrocarbon receptor interacts with ATP5alpha1, a subunit of the ATP synthase complex, and modulates mitochondrial function. AB - Dioxins, including 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD), produce a wide range of toxic effects in mammals. Most, if not all, of these toxic effects are regulated by the aryl hydrocarbon receptor (AHR). The AHR is a ligand activated transcription factor that has been shown to interact with numerous proteins capable of influencing the receptor's function. The ability of secondary proteins to alter AHR-mediated transcriptional events, a necessary step for toxicity, led us to determine whether additional interacting proteins could be identified. To this end, we have employed tandem affinity purification (TAP) of the AHR in Hepa1c1c7 cells. TAP of the AHR, followed by mass spectrometry (MS) identified ATP5alpha1, a subunit of the ATP synthase complex, as a strong AHR interactor in the absence of ligand. The interaction was lost upon exposure to TCDD. The association was confirmed by co-immunoprecipitation in multiple cell lines. In addition, cell fractionation experiments showed that a fraction of the AHR is found in the mitochondria. To ascribe a potential functional role to the AHR:ATP5alpha1 interaction, TCDD was shown to induce a hyperpolarization of the mitochondrial membrane in an AHR-dependent and transcription-independent manner. These results suggest that a fraction of the total cellular AHR pool is localized to the mitochondria and contributes to the organelle's homeostasis. PMID- 21616092 TI - And yet it moves: perceptual illusions and neural mechanisms of pursuit compensation during smooth pursuit eye movements. AB - Pursuit eye movements are smooth rotations of the eye aimed at tracking moving objects. During pursuit, the visual system "compensates" for the eye movements, and transforms image movements captured by the eye from retinal to extra-retinal coordinates, for world-centered perception and action. When this function is impaired such as in schizophrenia, subjects misattribute retinal movements generated by their own eye movements to external sources. Surprisingly, even in healthy subjects pursuit compensation is incomplete, and results in illusory perception of motion. Neurophysiological, psychophysical and imaging studies elucidated many aspects of the neural substrates of visual processing during pursuit, including where and how in the cortex visual and non-visual signals interact to produce extra-retinal perception of motion. Here we review current understanding of motion processing in the visual cortex during pursuit and its relation to perception, from a broad perspective drawing from electrophysiology, fMRI, psychophysics and computational modeling. We discuss the experimental findings in the context of theories of pursuit compensation, and review some of the open questions in the field. PMID- 21616093 TI - In vitro production of Helicoverpa baculovirus biopesticides--automated selection of insect cell clones for manufacturing and systems biology studies. AB - Baculovirus pesticides are increasingly being used as effective biological control agents against caterpillar pests worldwide. Increasing occlusion body (OB) yields per cell in culture is the main challenge to enable commercialization of in vitro production of baculovirus pesticides. Isolating clones from a heterogeneous cell population may allow development of a high virus producing cell clone. To date, the selection of insect clones has been based mainly on laborious cell serial dilution methods which create few viable clones. This work used an automated robotic clone picking system to establish over 250 insect clones of a Helicoverpa zea cell population to be screened for virus production. However, the higher producing clones only produced 10-30% higher OB yields than the original cell population. This study suggested that unless screening of thousands of clones is performed, obtaining a 2-fold increase in OB/cell yield compared to the parent population is unlikely. Nevertheless, it creates pure clones for manufacturing. In addition, two clones that were at least 2-3 times different in OB yields were isolated. Hence, this method can create a high contrast system (OB/cell yield basis), for comparative studies using a systems biology approach, which should inform a more targeted approach to engineer genetically a production cell line. PMID- 21616094 TI - Depletion of GTP pool is not the predominant mechanism by which ribavirin exerts its antiviral effect on Lassa virus. AB - Ribavirin (1-beta-d-ribofuranosyl-1,2,4-triazole-3-carboxamide) is the standard treatment for Lassa fever, though its mode of action is unknown. One possibility is depletion of the intracellular GTP pool via inhibition of the cellular enzyme inosine monophosphate dehydrogenase (IMPDH). This study compared the anti arenaviral effect of ribavirin with that of two other IMPDH inhibitors, mycophenolic acid (MPA) and 5-ethynyl-1-beta-d-ribofuranosylimidazole-4 carboxamide (EICAR). All three compounds were able to inhibit Lassa virus replication by >=2 log units in cell culture. Restoring the intracellular GTP pool by exogenous addition of guanosine reversed the inhibitory effects of MPA and EICAR, while ribavirin remained fully active. Analogous experiments performed with Zaire Ebola virus showed that IMPDH inhibitors are also active against this virus, although to a lesser extent than against Lassa virus. In conclusion, the experiments with MPA and EICAR indicate that replication of Lassa and Ebola virus is sensitive to depletion of the GTP pool mediated via inhibition of IMPDH. However, this is not the predominant mechanism by which ribavirin exerts its in vitro antiviral effect on Lassa virus. PMID- 21616095 TI - Significantly increased CCL5/RANTES and CCR7 mRNA levels in localized scleroderma. AB - Plaque-type morphea, the most common subtype of localized scleroderma (LS), is a connective tissue disease which is characterized by immunological dysregulation, vascular alterations, and skin fibrosis. We aimed to investigate the expression profiles of different cytokines and chemokines in patients with LS and healthy controls. Twenty patients with plaque-type morphea and 18 healthy controls were investigated. Skin biopsies were performed for real-time RT-PCR studies. Median mRNA of interleukin (IL)-6 was significantly higher expressed in LS than in normal skin. By contrast, median IL-1alpha mRNA levels were significantly decreased in LS as compared to controls. Median mRNA expression of CCL13, IL 1beta, IL-2, and tumor necrosis factor-alpha did not significantly differ between LS lesions and healthy skin. However, we observed significantly increased median chemokine ligand 5/RANTES (CCL5/RANTES) and median chemokine receptor 7 (CCR7) mRNA expression in LS lesions as compared to healthy controls. CCL5/RANTES and CCR7 mRNA expression significantly correlated in LS lesions. We could confirm data of previous studies indicating that gene expression of IL-6 is up-regulated in LS lesions as compared to healthy skin. Moreover, we have shown for the first time a significant increase of mRNA levels of CCR7 and CCL5/RANTES in LS lesions indicating an important pathogenetic role of chemokines in LS. PMID- 21616096 TI - Altered regulation of renal nitric oxide and atrial natriuretic peptide systems in angiotensin II-induced hypertension. AB - The present study was aimed to determine whether there is an altered role of local nitric oxide (NO) and atrial natriuretic peptide (ANP) systems in the kidney in association with the angiotensin (Ang) II-induced hypertension. Male Sprague-Dawley rats were used. Ang II (100 ng.min-1.kg-1) was infused through entire time course. Thirteenth day after beginning the regimen, kidneys were taken. The protein expression of NO synthase (NOS) and nitrotyrosine was determined by semiquantitative immunoblotting. The mRNA expression of components of ANP system was determined by real-time polymerase chain reaction. The activities of soluble and particulate guanylyl cyclases were determined by the amount of cGMP generated in responses to sodium nitroprusside and ANP, respectively. There developed hypertension and decreased creatinine clearance in the experimental group. The protein expression of eNOS, nNOS and nitrotyrosine was increased in the cortex, while that of iNOS remained unaltered. The urinary excretion of NO increased in Ang II-induced hypertensive rats. The catalytic activity of soluble guanylyl cyclase was blunted in the glomerulus in Ang II induced hypertensive rats. The mRNA expression of ANP was increased in Ang II induced hypertensive rats. Neither the expression of NPR-A nor that of NPR-C was changed. The protein expression of neutral endopeptidase was decreased and the activity of particulate guanylyl cyclase was blunted in the glomerulus and papilla in Ang II-induced hypertensive rats. In conclusion, the synthesis of NO and ANP was increased in the kidney of Ang II-induced hypertension, while stimulated cGMP response was blunted. These results suggest desensitization of guanylyl cyclase in the kidney of Ang II-induced hypertensive rats, which may contribute to the associated renal vasoconstriction and hypertension. PMID- 21616097 TI - Release of podocalyxin into the extracellular space. Role of metalloproteinases. AB - Podocalyxin (PODXL) is a type I membrane mucoprotein abundantly presented in the epithelial cells (podocytes) of kidney glomeruli where it plays an important role in maintaining the plasma filtration. PODXL is also expressed in other types of cells but its function is ignored. A recombinant soluble fragment of the PODXL ectodomain modifies the signaling of the membrane bound PODXL. Based on this antecedent, we aimed at investigating whether PODXL could be cleaved and released into the extracellular space as a soluble peptide. In this study, we used a fusion protein of human PODXL and green fluorescent protein expressed in CHO cells (CHO-PODXL-GFP) and a human tumor cell (Tera-1) inherently expressing PODXL. PODXL was detected by wide-field microscopy in the Golgi, the plasma membrane and in a vesicular form preferentially located at the leading edges of the cell and also progressing along the filopodium. We detected PODXL in the insoluble and soluble fractions of the extracellular medium of CHO-PODXL-GFP cells. Stimulation of protein kinase C (PKC) by Phorbol-12-myristate-13-acetate (PMA) enhanced the release of PODXL to the extracellular space whereas this effect was prevented either by inhibitors of PKC or specific inhibitors of matrix metalloproteinases. It is concluded that intact PODXL is released to the extracellular space as a cargo of microvesicles and also as a soluble cleaved fragment of ectodomain. PMID- 21616099 TI - Viable but non-culturable (VBNC) bacteria: Gene expression in planktonic and biofilm cells. AB - Viable but non-culturable (VBNC) bacteria are common in nutrient poor and/or stressed environments as planktonic cells and biofilms. This article discusses approaches to researching VBNC bacteria to obtain knowledge that is lacking on their gene expression while in the VBNC state, and when they enter into and then recover from this state, when provided with the necessary nutrients and environmental conditions to support growth and cell division. Two-dimensional gel electrophoresis of proteins, global gene expression, reverse-transcription polymerase chain reaction (PCR) analysis and sequencing by synthesis coupled with data on cell numbers, viability and species present are central to understanding the VBNC state. PMID- 21616098 TI - Proteolytic action of kallikrein-related peptidase 7 produces unique active matrix metalloproteinase-9 lacking the C-terminal hemopexin domains. AB - The gelatinases, matrix metalloproteinase (MMP)-9 and -2, are produced as latent, inactive enzymes that can be proteolytically activated by a number of proteases. In many normal and pathological conditions, where the expression of MMPs is deregulated, changes in the expression of other proteases have also been reported. Human kallikrein-related peptidase 7 (KLK7), a chymotryptic-like serine protease, is overexpressed in many different types of neoplastic conditions, which have also been shown to express high levels of both MMP-9 and -2. Since the activation of MMPs by KLK7 has never been examined, we sought to determine whether KLK7 can activate these MMPs. To test this hypothesis KLK7 was incubated with the recombinant MMPs and the products of the reaction were analyzed for their activity. Incubation of proMMP-9 with KLK7 resulted in the production of a novel truncated, active MMP-9 lacking the C-terminal hemopexin domains. In contrast, KLK7 degraded, but did not activate, proMMP-2. The novel activation of proMMP-9 by KLK7 was further confirmed using conditioned medium prepared from an MMP-9-expressing cell line, MDA-MMP-9. Our results clearly establish that KLK7 activates proMMP-9 to produce a novel truncated, active MMP-9 product not generated by other proteases. These findings suggest that KLK7 may play an important role in the activation of MMP-9 in tumors that express high levels of both these proteases and the resulting truncated MMP may possess altered substrate specificities compared with full-length MMP-9 activated by other proteases. PMID- 21616100 TI - Proline repeats, in cis- and trans-positions, confer protection against the toxicity of misfolded proteins in a mammalian cellular model. AB - A broad range of neurodegenerative disorders result from the cytotoxicity conferred by aberrantly folded mutant proteins. Intriguingly, the cytotoxicity and aggregation property of a few mutant proteins are known to be modulated by the flanking sequences. One of such modulators is the proline repeat tract. Using a mammalian cellular model, we show here that proline repeat tract, both in cis- and in trans-positions, ameliorate the cytotoxicity of wide range of misfolded proteins coded by synthetic constructs. We further show that the proline repeat tract could possibly confer protection against the cytotoxicity of misfolded proteins by altering their conformation at the time of their synthesis. Thus, our study elucidates the mechanism by which the proline repeat tract might ameliorate the toxicity of misfolded proteins, and opens up new therapeutic modalities for disorders caused by cytotoxic misfolded proteins. PMID- 21616101 TI - Role of activating transcription factor 3 in ischemic penumbra region following transient middle cerebral artery occlusion and reperfusion injury. AB - The activating transcription factor 3 (ATF3) is expressed by various types of cellular insults. It has been suggested to serve diverse functions in both cellular survival and death signal cascades, but the exact role of ATF3 in brain ischemia is little known so far. Thus, the authors examined the expression pattern of ATF3 following middle cerebral artery occlusion (MCAO) and reperfusion injury. At 1-2 days after MCAO and reperfusion injury, numerous number of ATF3 immunoreacitive (-ir) nuclei was observed in the ipsilateral peri-infarct cortex, but declined rapidly at 3 days. Almost all ATF3-ir nuclei were co-localized with NeuN-ir neurons. Neither GFAP- nor OX42-ir neuroglia were co-localized with ATF3. Double labeling of Fluoro-Jade B with ATF3 showed that ATF3-ir nuclei mismatched with Fluoro-Jade B-ir neurons. To further examine the role of ATF3 in ischemic peri-infarct regions, double immunofluorescent labeling of ATF3/caspase 3, ATF3/Bcl-xl, and ATF3/HSP27 was conducted. Semiquantitive estimation showed that about 15% of ATF3-ir neurons also expressed caspase 3. However, about only 0.4% and 2.6% of ATF3-ir neurons were double-stained with Bcl-xl and Hsp27, respectively. Consequently, it would be suggested that ATF3 seem to play an important role in caspase-dependent neuronal apoptotic signal transduction pathways caused by focal cerebral ischemia and reperfusion injury. PMID- 21616102 TI - PhoN-expressing, lyophilized, recombinant Deinococcus radiodurans cells for uranium bioprecipitation. AB - Employment of genetically engineered radiation resistant organisms to recover radionuclides/heavy metals from radioactive wastes is an attractive proposition. Cells of recombinant Deinococcus radiodurans strain expressing, a non-specific acid phosphatase encoding phoN gene, were lyophilized. Lyophilized recombinant Deinococcus cells retained viability and PhoN activity and could efficiently precipitate uranium from aqueous solutions for up to six months of storage at room temperature. Batch process for uranium removal using lyophilized cells was more efficient compared to a flow through system, in terms of percent uranium removed, substrate conservation and time taken. Lyophilized recombinant Deinococcus cells exhibited high loading of up to 5.7 g uranium/g dry weight of cells in a batch process at 20 mM input uranium concentration. Lyophilization deflated the cells but did not alter gross cell morphology or surface nucleation capability of cells for uranium precipitation. The precipitated uranyl phosphate remained tightly associated with the cell surface, thus facilitating easy recovery. PMID- 21616104 TI - Kinetics of transport of doxorubicin bound to nanoparticles across the blood brain barrier. AB - Drug delivery to the brain is restricted due to the blood-brain barrier (BBB). Previously, it has been shown that surfactant-coated doxorubicin-loaded nanoparticles were successful in overcoming the BBB and were effective in the treatment of rat brain tumours. However, drug distribution in brain tissue after crossing the BBB was never determined. To distinguish between the amounts of drug in the whole brain and the fraction of drug in the brain parenchyma after crossing the BBB a capillary depletion technique was employed. For this purpose rats were intravenously treated with a doxorubicin solution in 1% polysorbate 80, or doxorubicin-loaded poly-(n-butyl cyanoacrylate) (PBCA) nanoparticles without and with 1% polysorbate 80 coating, respectively. The dosage of doxorubicin was 5 mg per kg of rat body weight. At 30 min, 2 h, and 4 h following intravenous injection into the tail vein, the rats were sacrificed and their brains removed. Homogenates of the brains were prepared. In addition, one part of the homogenate was separated by centrifugation into a pellet (vascular elements) and supernatant (parenchyma) using a well established capillary depletion technique. The time dependent distribution of doxorubicin in these brain fractions was studied. Clinically effective concentrations in all investigated brain fractions could only be detected in rats treated with surfactant-coated nanoparticles, indicating a significant transcytosis across the BBB. Only low concentrations were observed after 0.5 and 2 h with the uncoated nanoparticles. No uptake of doxorubicin into the brain was observable after administration of drug solution alone. These observations demonstrate the great potential of surface-coated PBCA nanoparticles for the delivery of drugs to the central nervous system. PMID- 21616105 TI - A first step towards a mouse model for hepatitis C virus infection containing a human immune system. AB - BACKGROUND & AIMS: Studies of hepatitis C virus (HCV) infection, immunopathogenesis, and resulting liver diseases have been hampered by the lack of a small animal model. We developed humanized mice with human immune system and liver tissues to improve the studies of hepatitis C pathogenesis and treatment. METHODS: To promote engraftment of human hepatocytes, we expressed a fusion protein of the FK506 binding protein (FKBP) and caspase 8 under the control of the albumin promoter (AFC8), which induces liver cell death, in Balb/C Rag2(-/-) gammaC-null mice. Co-transplantation of human CD34(+) human hematopoietic stem cells (HSC) and hepatocyte progenitors into the transgenic mice led to efficient engraftment of human leucocytes and hepatocytes. We then infected these humanized mice (AFC8-hu HSC/Hep) with primary HCV isolates and studied HCV-induced immune responses and liver diseases. RESULTS: AFC8-hu HSC/Hep mice supported HCV infection in the liver and generated a human immune T-cell response against HCV. HCV infection induced liver inflammation, hepatitis, and fibrosis, which correlated with activation of stellate cells and expression of human fibrogenic genes. CONCLUSIONS: AFC8-hu HSC/Hep mice are a useful model of HCV infection, the immune response, and liver disease, because they contain human immune system and liver cells. These mice become infected with HCV, generate a specific immune response against the virus, and develop liver diseases that include hepatitis and fibrosis. This model might also be used to develop therapeutics for HCV infection. PMID- 21616106 TI - Depicting the role of TP53 in hepatocellular carcinoma progression. AB - BACKGROUND & AIMS: Mutations in TP53, a tumor suppressor gene, are associated with prognosis of many cancers. However, the prognostic values of TP53 mutation sites are not known for patients with hepatocellular carcinoma (HCC) because of heterogeneity in their geographic and etiologic backgrounds. METHODS: TP53 mutations were investigated in a total of 409 HCC patients, including Chinese (n=336) and white (n=73) patients, using the direct sequencing method. RESULTS: A total of 125 TP53 mutations were found in Chinese patients with HCC (37.2%). HCC patients with TP53 mutations had a shorter overall survival time compared with patients with wild-type TP53 (hazard ratio [HR], 1.86; 95% confidence interval [CI]: 1.37-2.52; P<.001). The hot spot mutations R249S and V157F were significantly associated with worse prognosis in univariate (HR, 2.11; 95% CI: 1.51-2.94; P<.001) and multivariate analyses (HR, 1.79; 95% CI: 1.29-2.51; P<.001). Gene expression analysis revealed the existence of stem cell-like traits in tumors with TP53 mutations. These findings were validated in breast and lung tumor samples with TP53 mutations. CONCLUSIONS: TP53 mutations, particularly the hot spot mutations R249S and V157F, are associated with poor prognosis for patients with HCC. The acquisition of stem cell-like gene expression traits might contribute to the aggressive behavior of tumors with TP53 mutation. PMID- 21616107 TI - Lactose characteristics and the generation of the aerosol. AB - The delivery efficiency of dry-powder products for inhalation is dependent upon the drug formulation, the inhaler device, and the inhalation technique. Dry powder formulations are generally produced by mixing the micronised drug particles with larger carrier particles. These carrier particles are commonly lactose. The aerosol performance of a powder is highly dependent on the lactose characteristics, such as particle size distribution and shape and surface properties. Because lactose is the main component in these formulations, its selection is a crucial determinant of drug deposition into the lung, as interparticle forces may be affected by the carrier-particle properties. Therefore, the purpose of this article is to review the various grades of lactose, their production, and the methods of their characterisation. The origin of their adhesive and cohesive forces and their influence on aerosol generation are described, and the impact of the physicochemical properties of lactose on carrier-drug dispersion is discussed in detail. PMID- 21616108 TI - Viral delivery for gene therapy against cell movement in cancer. AB - Viral delivery for cancer gene therapy is a promising approach, where traditional radiotherapy or chemotherapy to limit proliferation and movement of cancer cells has met resistance. Based on the new understanding of the biology of the viral vectors, therapeutic viral vectors for cancer gene therapy have been improved for greater safety and efficacy as well as transitioned from being non-replicating to replication-competent. Traditional oncolytic vectors have focused on eliminating tumor growth, while novel vectors simultaneously target epithelial-to-mesenchymal transition (EMT) in cancer cells, which could further prevent and reverse the aggressive tumor progression. In this review, we highlight the illustrative examples of cancer gene therapy in clinical trials as well as preclinical data and include proposals on methods to further enhance the safety and efficacy of oncolytic viral vectors in cancer gene therapy. PMID- 21616109 TI - Stimulatory effect of intracerebroventricular administration of orexin A on food intake in the zebrafish, Danio rerio. AB - Orexin is a potent orexigenic neuropeptide implicated in feeding regulation of mammals. However, except for the case of goldfish, the involvement of orexin in the feeding behavior of teleost fish has not well been studied. Therefore, we investigated the role of orexin on food intake using a zebrafish (Danio rerio) model. We examined the effect of feeding status on orexin-like immunoreactivity and the expression level of orexin transcript in the brain. The number of neuronal cells showing orexin-like immunoreactivity in the hypothalamic region, including the posterior tuberal nucleus, was significantly increased in fish fasted for 7days. Orexin precursor mRNA levels in the brain obtained from fish fasted for 7 days were higher than those in fish that had been fed normally. We then investigated the effect of intracerebroventricular (ICV) administration of orexin A on food intake. Cumulative food intake was significantly increased by ICV administration of orexin A (at 0.3 and 3 pmol/g body weight, BW) during a 60 min observation period after treatment. The orexin A-induced orexigenic action (at 0.3 pmol/g BW) was blocked by treatment with an orexin receptor antagonist, SB334867, at 10 pmol/g BW. These results indicate that orexin A acts as feeding regulator in the zebrafish. PMID- 21616110 TI - Effects of cell-type specific leptin receptor mutation on leptin transport across the BBB. AB - The functions of leptin receptors (LRs) are cell-type specific. At the blood brain barrier, LRs mediate leptin transport that is essential for its CNS actions, and both endothelial and astrocytic LRs may be involved. To test this, we generated endothelia specific LR knockout (ELKO) and astrocyte specific LR knockout (ALKO) mice. ELKO mice were derived from a cross of Tie2-cre recombinase mice with LR-floxed mice, whereas ALKO mice were generated by a cross of GFAP-cre with LR-floxed mice, yielding mutant transmembrane LRs without signaling functions in endothelial cells and astrocytes, respectively. The ELKO mutation did not affect leptin half-life in blood or apparent influx rate to the brain and spinal cord, though there was an increase of brain parenchymal uptake of leptin after in situ brain perfusion. Similarly, the ALKO mutation did not affect blood brain barrier permeation of leptin or its degradation in blood and brain. The results support our observation from cellular studies that membrane-bound truncated LRs are fully efficient in transporting leptin, and that basal levels of astrocytic LRs do not affect leptin transport across the endothelial monolayer. Nonetheless, the absence of leptin signaling at the BBB appears to enhance the availability of leptin to CNS parenchyma. The ELKO and ALKO mice provide new models to determine the dynamic regulation of leptin transport in metabolic and inflammatory disorders where cellular distribution of LRs is shifted. PMID- 21616111 TI - Upregulation of KIR3DL1 gene expression in intestinal mucosa in active celiac disease. AB - Killer cell immunoglobulin-like receptors (KIRs) modulate natural killer (NK) and T-cell function by human leukocyte antigen class I interaction and have been implicated in celiac disease (CD). Qualitative expression of 16 KIR genes was determined in biopsies from 22 CD patients at diagnosis and after >2 years on a gluten-free diet (GFD). Quantitative expression analysis of KIR2DL4, KIR3DL1, KIR3DL3, and KLRC2 (a marker of an NK-reprogrammed T-cell subpopulation augmented in CD) was performed in 35 additional CD biopsy pairs and 14 non-CD control biopsies. No specific KIR expression profile was observed in CD. KIR3DL1 was more frequently expressed in active CD compared with GFD (p = 0.0312) and controls (p = 0.0008), with slightly increased levels in active disease. KLRC2 was overexpressed in active (p = 0.0037) and GFD (p = 0.0469) patients compared with non-CD controls and coexpressed with KIR3DL1. Results suggest the participation of KIR3DL1 overexpression in the overall immune activation seen in CD mucosa, which could be partly explained by the NK-like T-cell subpopulation increase. PMID- 21616113 TI - Towards ambient temperature-stable vaccines: the identification of thermally stabilizing liquid formulations for measles virus using an innovative high throughput infectivity assay. AB - As a result of thermal instability, some live attenuated viral (LAV) vaccines lose substantial potency from the time of manufacture to the point of administration. Developing regions lacking extensive, reliable refrigeration ("cold-chain") infrastructure are particularly vulnerable to vaccine failure, which in turn increases the burden of disease. Development of a robust, infectivity-based high throughput screening process for identifying thermostable vaccine formulations offers significant promise for vaccine development across a wide variety of LAV products. Here we describe a system that incorporates thermal stability screening into formulation design using heat labile measles virus as a prototype. The screening of >11,000 unique formulations resulted in the identification of liquid formulations with marked improvement over those used in commercial monovalent measles vaccines, with <1.0 log loss of activity after incubation for 8h at 40 degrees C. The approach was shown to be transferable to a second unrelated virus, and therefore offers significant promise towards the optimization of formulation for LAV vaccine products. PMID- 21616114 TI - Human papillomavirus (HPV) vaccination: perception and practice among French general practitioners in the year since licensing. AB - Acceptance of the Human Papillomavirus (HPV) vaccine by targeted populations will depend to a large extent on its acceptability among physicians. We examined the perceptions, attitudes and practices of general practitioners (GPs) in relation to HPV vaccination. From November 2007 to April 2008, a cross-sectional survey was carried out among a representative 5% sample of GPs in the large Rhone-Alpes region of France. Both quantitative (self-administered questionnaire) and qualitative (interview) approaches were used. During the month preceding the survey, 75.6% of the 279 GPs who responded had given at least one HPV vaccination and 47.6% had given a vaccination at the routine target age of 14 years. Overall, 80.8% of GPs reported a favourable opinion about HPV vaccination, 17.4% were uncertain and 1.8% were opposed. The main justification for a favourable opinion related to the public health benefits of the HPV vaccination (cited by 60% of those favouring vaccination). The main justification for an "opposed or uncertain" opinion was the too recent introduction of the vaccine (cited by 43.4%). The major difficulties in providing HPV vaccination were patients' concerns about potential side effects (cited by 37% of the respondents) and the target age of 14 years (28.9%). Interviews suggested that the concern about age may relate to the need, as perceived by GPs, to discuss sexually transmitted infections with adolescent patients. A favourable opinion about HPV vaccination was associated with seeing more female patients per week, younger age, and GPs' intention to recommend hepatitis B vaccination. This representative survey of GPs in a major region of France finds a favourable opinion about the HPV vaccine and widespread use of it, despite some concerns that the recent introduction of the vaccine means that we do not yet fully understand the potential for side effects and about the recommended target age of recipients. PMID- 21616115 TI - Evidence of an effect of BCG revaccination on incidence of tuberculosis in school aged children in Brazil: second report of the BCG-REVAC cluster-randomised trial. AB - BCG revaccination is still used in some tuberculosis endemic countries. Until now, the little evidence available suggested that BCG revaccination confers very limited additional protection, although there was no information on whether protection depends on the setting and age of revaccination, or if protection increases with time since vaccination. Here we report on an extended follow up of the BCG-REVAC trial, a cluster randomised trial conducted in the Brazilian cities Salvador and Manaus including over 200,000 children aged 7-14 years aimed to evaluate the efficacy of BCG revaccination in children who had received neonatal BCG vaccination. With the extended follow-up (9 years) and the additional cases accrued we now have enough power to report vaccine efficacy separately for the two cities (with different distances from Equator and presumably different prevalence of non-tuberculosis mycobacteria), and by age at vaccination and clinical form. The overall vaccine efficacy was 12% (-2 to 24%) as compared to 9% (-16 to 29%) for the 5-year follow up. Vaccine efficacy was higher in Salvador (19%, 3 to 33%) than in Manaus (1%, -27 to 27%) with the highest vaccine efficacy in children from Salvador aged <11 years at revaccination (33%, 3 to 54%). The findings are in line with the hypothesis that BCG vaccination offers higher efficacy in low NTMb prevalence, and show that revaccination with BCG can offer weak protection in selected subgroups. PMID- 21616112 TI - Mouse models to assess the efficacy of non-typhoidal Salmonella vaccines: revisiting the role of host innate susceptibility and routes of challenge. AB - Non-typhoidal Salmonella enterica (NTS) serovars Typhimurium and Enteritidis are important causes of bacterial gastroenteritis in the USA and worldwide. In sub Saharan Africa these two serovars are emerging as agents associated with lethal invasive disease (e.g., bacteremia, meningitis). The development of NTS vaccines, based on mucosally administered live attenuated strains and parenteral non-living antigens, could diminish the NTS disease burden globally. Mouse models of S. Typhimurium and S. Enteritidis invasive disease can accelerate the development of NTS vaccines. Live attenuated NTS vaccines elicit both cellular and humoral immunity in mice and their efficacy is well established. In contrast, non-living vaccines that primarily elicit humoral immunity have demonstrated variable efficacy. An analysis of the reported studies with non-living vaccines against S. Typhimurium and S. Enteritidis reveals that efficacy is influenced by two important independent variables: (1) the innate susceptibility to NTS infection that differs dramatically between commonly used mouse strains and (2) the virulence of the NTS strain used for challenge. Protection by non-living vaccines has generally been seen only in host-pathogen interactions where a sub-lethal infection results, such as challenging resistant mice with either highly virulent or weakly virulent strains or susceptible mice with weakly virulent strains. The immunologic basis of this discrepancy and the implications for human NTS vaccine development are reviewed herein. PMID- 21616116 TI - A systematic review of experimental infections with enterotoxigenic Escherichia coli (ETEC). AB - Volunteer challenge with enterotoxigenic Escherichia coli (ETEC) has been used for four decades to elucidate the pathogenesis and immune responses and assess efficacy of various interventions. We performed a systematic review of these studies and a meta-analysis of individual patient-level data (IPD) from a subset of studies using standard methodology. We identified 27 studies of 11 ETEC strains administered to 443 naive subjects at doses from 1*10(6) to 1*10(10) colony forming units (cfu). Diarrhea attack rates varied by strain, dose and enterotoxin. Similar rates were seen at doses of 5*10(8) to 1*10(10)cfu with the three most commonly used strains B7A, E24377A, H10407. In IPD analysis, the highest diarrhea attack rates were seen with strains B7A, H10407 and E24377A. The H10407 induced significantly higher stool output than the other strains. Additionally, the rate of output was different across strains. The risk of diarrhea, abdominal cramps, nausea and headaches differed significantly by ETEC strain. An increased risk of nausea, abdominal cramps and headaches was seen for females. Baseline anti-LT IgG titers appeared to be associated with a decrease risk of diarrhea outcomes, a trend not seen with anti-LT IgA or seen consistently with anti-colonization factor antibodies. Neither early antibiotic treatment nor diarrhea duration significantly affected the frequency or magnitude of serologic responses. These studies have served as an invaluable tool in understanding disease course, pathogenicity, innate immune responses and an early assessment of product efficacy. When designing and planning experimental ETEC infection studies in this age of increased ethical scrutiny and growing appreciation of post infectious sequelae, better understanding of available data is essential. PMID- 21616117 TI - Economic burden of HPV-related cancers in France. AB - Human papillomavirus (HPV) infection is associated with a range of diseases and cancers at different anatomical sites. In addition to its role as a necessary cause of cervical cancer, HPV is also associated with cancers of the vulva, vagina, anus, penis, head and neck. With the exception of cervical cancer, however, very few data are available on the economic burden of HPV-associated cancers. We assessed the annual costs associated with management of HPV-related cancers in France from the healthcare payers' perspective. We used data from studies that employed similar methodologies to estimate the costs during 2006 for cervical cancer, vulvar and vaginal cancers, anal cancer, and penile cancer, and during 2007 for head and neck cancers. Data on hospital-management costs for cancer were derived from the French national hospital database. The costs of outpatient care and daily allowance costs were estimated using data from the French National Institute of Cancer report for 2007. The costs for HPV-related cancers were estimated according to the percentage of each cancer type attributable to HPV infection. The estimated total costs associated with HPV related cancers in France were ?239.7 million. The overall costs in men were ?107.2 million, driven mainly by head and neck cancers (?94.6 million). The total costs in women were ?132.5 million, due mainly to invasive cervical cancer (?83.9 million). The costs associated with HPV-related cancers are important to consider when evaluating the overall benefits of HPV vaccination in males and females. PMID- 21616118 TI - Anterior cingulate proton spectroscopy glutamate levels differ as a function of smoking cessation outcome. AB - BACKGROUND: Cigarette smoking is the leading preventable cause of death. Unfortunately, the majority of smokers who attempt to quit smoking relapse within weeks. Abnormal dorsal anterior cingulate cortex (dACC) function may contribute to tobacco smoking relapse vulnerability. Growing evidence suggests that glutamate neurotransmission is involved in mediating nicotine dependence. We hypothesized that prior to a cessation attempt, dACC glutamate levels would be lower in relapse vulnerable smokers. METHODS: Proton magnetic resonance spectra (MRS) were obtained from dACC and a control region, the parieto-occipital cortex (POC), using two-dimensional J-resolved MRS at 4T and analyzed using LCModel. Nine nicotine-dependent women were scanned prior to making a quit attempt. Subjects then were divided into two groups; those able to maintain subsequent abstinence aided by nicotine replacement therapy (NRT) and those who slipped while on NRT (smoked any part of a cigarette after attaining at least 24h of abstinence). RESULTS: Slip subjects exhibited significantly reduced dACC MRS glutamate (Glu/Cr) levels (p<0.03) compared to abstinent subjects. This effect was not observed in the POC control region. CONCLUSIONS: Our preliminary findings suggest that dACC Glu levels as measured with MRS may help identify and/or be a biomarker for relapse vulnerable smokers. Future research following up on these findings may help clarify the role of dACC Glu in smoking dependence that may lead to new treatment strategies. PMID- 21616119 TI - Different role of Schisandrin B on mercury-induced renal damage in vivo and in vitro. AB - Mercuric chloride (HgCl2) causes acute oxidant renal failure that affects mainly proximal tubules. Schisandrin B (Sch B), an active lignan from the fruit of Schisandra chinensis, has been successfully used to treat gentamicin nephrotoxicity, but its role against mercury damage is still largely unknown. Here we analysed in vivo and in vitro the efficacy of Sch B supplementation against HgCl2 nephrotoxicity, focusing on histopathology, stress proteins, oxidative (cytochrome c oxidase) and nitrosactive markers (eNOS, nNOS). Wistar rats were treated with Sch B (10 mg/kg/day p.o.) or vehicle (olive oil) for 9 days, then coadministered with a single HgCl2 nephrotoxic dose (3.5 mg/kg i.p.) and killed after 24 h. The tubular and mitochondrial damage induced by mercury was limited by Sch B coadministration in vivo. Remarkably, after Sch B and mercury challenge, HSP25, HSP72, GRP75 were reduced in the renal cortex, cytochrome c oxidase increased and eNOS and nNOS were restored in glomeruli. In contrast, NRK-52E proximal tubular cells treated with Sch B 6.25 MUM plus HgCl2 20 MUM did not show any amelioration on viability and oxidative stress in respect to HgCl2 20 MUM alone. Moreover, after Sch B plus mercury in vitro treatment, HSP72 staining persisted while HSP25 further increased. Thus, in our experimental conditions, Sch B cotreatment afforded better protection against mercury poisoning in vivo than in vitro. This discrepancy might be partly attributable to Sch B influence on glomerular perfusion as corroborated by the recovery of vasoactive markers like macular and endothelial nitric oxide isoforms. PMID- 21616120 TI - Ghrelin and cancer. AB - Ghrelin is a peptide hormone that was originally isolated from the stomach as the endogenous ligand for the growth hormone secretagogue receptor (GHSR). Ghrelin has many functions, including the regulation of appetite and gut motility, growth hormone release from the anterior pituitary and roles in the cardiovascular and immune systems. Ghrelin and its receptor are expressed in a number of cancers and cancer cell lines and may play a role in processes associated with cancer progression, including cell proliferation, apoptosis, and cell invasion and migration. PMID- 21616122 TI - Ghrelin axis genes, peptides and receptors: recent findings and future challenges. AB - The ghrelin axis consists of the gene products of the ghrelin gene (GHRL), and their receptors, including the classical ghrelin receptor GHSR. While it is well known that the ghrelin gene encodes the 28 amino acid ghrelin peptide hormone, it is now also clear that the locus encodes a range of other bioactive molecules, including novel peptides and non-coding RNAs. For many of these molecules, the physiological functions and cognate receptor(s) remain to be determined. Emerging research techniques, including proteogenomics, are likely to reveal further ghrelin axis-derived molecules. Studies of the role of ghrelin axis genes, peptides and receptors, therefore, promises to be a fruitful area of basic and clinical research in years to come. PMID- 21616121 TI - Characterization of murine melanocortin receptors mediating adipocyte lipolysis and examination of signalling pathways involved. AB - The melanocortin receptors (MCRs) belong to the G-protein coupled receptors (family A). So far, 5 different subtypes have been described (MC1R-MC5R) and of these MC2R and MC5R have been proposed to act directly in adipocytes and regulate lipolysis in rodents. Using ACTH and alpha-melanocyte stimulating hormone (alpha MSH) generated from proopiomelanocortin (POMC), as well as synthetic MSH analogues to stimulate lipolysis in murine 3T3-L1 adipocytes it is shown that MC2R and MC5R are lipolytic mediators in differentiated 3T3-L1 adipocytes. Involvement of cAMP, phosphorylated extracellular signal-regulated kinase (ERK) 1/2, protein kinase B (PKB), adenosine 5' monophosphate activated protein kinase (AMPK) and Jun-amino-terminal kinase (JNK) in MCR mediated lipolysis were studied. Interestingly, results obtained in 3T3-L1 cells suggest that lipolysis stimulated by alpha-MSH, NDP-alpha-MSH, MT-II, SHU9119 and PG-901 is mediated through MC5R in a cAMP independent manner. Finally, we identify essential differences in MCR mediated lipolysis when using 3T3-L1 cells compared to primary adipocytes. PMID- 21616123 TI - Regulator of G protein signaling 9-2 (RGS9-2) mRNA is up regulated during neuronal differentiation of mouse embryonic stem cells. AB - In this study we demonstrate up-regulation of mRNA for Regulator of G protein Signaling (RGS) 6, 7, 9 and 11, R7 family RGS binding protein (R7BP) and RGS9 anchor protein (R9AP) during neuronal differentiation of mouse embryonic stem cells (mESCs). This expression pattern was most robust for RGS9 whose transcript level was low in undifferentiated mESCs but increased over 125 fold when differentiating mESCs began to exhibit a neuronal precursor cell (NPC) phenotype. In addition, we demonstrate that RGS9 mRNA is expressed in neuronal stem cells isolated from embryonic mouse cortex. The expression of RGS9 in two distinct populations of NPCs suggests that RGS9 and its accessory proteins may play an important role in neuron development. PMID- 21616124 TI - Reversibility in Chinese word formation influences target identification. AB - We recorded event-related brain potentials during the processing of visually presented compound words in Mandarin Chinese. We capitalized on a particular characteristic of Chinese word formation, where two constituents can be combined in two different orders (A+B or B+A), yielding distinct meanings-so-called "reversible words". By investigating the impact of structural reversibility on the processing of compounds in Chinese during a lexical decision task, the present study revealed a pronounced difference between reversible and non reversible words. Analyses revealed a more enhanced negativity (N400) for reversible words, reflecting demands during semantic processing, followed by a P300-like deflection for non-reversible words, which we interpret with respect to the ease of target identification due to target discriminability. The findings indicate that both character combinations (A+B, B+A) as well as access to the individual constituent meanings result in interference during the processing of reversible and non-reversible words. They reveal that combinatorial processes are affected by demands arising from competing representations (N400), and more importantly, that this impacts the task-relevant cognitive processes required in the current word/non-word identification task (P300). The observed P300-like potential suggests that the allocation of attentional resources is intertwined with combinatorial operations. PMID- 21616125 TI - Subventricular zone in motor neuron disease with frontotemporal dementia. AB - Investigate how the subventricular proliferation and organisation is modified in a patient with FTLD-ALS. We studied the subventricular zone (SVZ) of a patient with FTLD-ALS immunohistochemical and histologically. We found an increase of Ki 67 positive cells and neuroblast in the subventricular zone, suggesting an activation of proliferating activity in response to FTD-ALS. This proliferation can act as a compensatory mechanism for rapid neuronal death and its modulation could provide a new therapeutic pathway in ALS. These results suggest a modification of neurogenesis in FTD-ALS. PMID- 21616126 TI - Role of apoptosis inducing factor (AIF) for hippocampal neuronal cell death following global cerebral ischemia in mice. AB - The molecular mechanisms of neuronal cell death following circulatory arrest are still not fully understood. In the current study we investigated the role of apoptosis-inducing factor (AIF), a major caspase-independent mitochondrial cell death protein, for neuronal cell death following global cerebral ischemia (GCI). C57/Bl6 or low AIF expressing Harlequin mutant mice (AIF(low)) and their wild type littermates were subjected to 10 min of GCI. DNA damage, nuclear pathology, and localization of AIF were investigated 6, 24, and 72 h after GCI by TUNEL and DAPI staining, and immunohistochemistry, respectively. Cell death of hippocampal CA1 neurons following GCI was associated with nuclear translocation of AIF, nuclear pyknosis, and DNA fragmentation, i.e. ~80% of all TUNEL-positive neurons had nuclear AIF staining. In AIF(low) mice neuronal cell loss was reduced by 60% (p<0.02). The current experiments suggest that AIF-mediated signaling represents a novel mechanism of neuronal cell death following GCI. PMID- 21616127 TI - The neuronal influence on tumor progression. AB - Nerve fibers accompany blood and lymphatic vessels all over the body. An extensive amount of knowledge has been obtained with regard to tumor angiogenesis and tumor lymphangiogenesis, yet little is known about the potential biological effects of "neoneurogenesis". Cancer cells can exploit the advantage of the factors released by the nerve fibers to generate a positive microenvironment for cell survival and proliferation. At the same time, they can stimulate the formation of neurites by secreting neurotrophic factors and axon guidance molecules. The neuronal influence on the biology of a neoplasm was initially described several decades ago. Since then, an increasing amount of experimental evidence strongly suggests the existence of reciprocal interactions between cancer cells and nerves in humans. Moreover, researchers have been able to demonstrate a crosstalk between cancer cells and nerve fibers as a strategy for survival. Despite all these evidence, a lot remains to be done in order to clarify the role of neurotransmitters, neuropeptides, and their associated receptor-initiated signaling pathways in the development and progression of cancer, and response to therapy. A global-wide characterization of the neurotransmitters or neuropeptides present in the tumor microenvironment would provide insights into the real biological influences of the neuronal tissue on tumor progression. This review is intended to discuss our current understanding of neurosignaling in cancer and its potential implications on cancer prevention and therapy. The review will focus on the soluble factors released by cancer cells and nerve endings, their biological effects and their potential relevance in the treatment of cancer. PMID- 21616128 TI - Parvalbumin interneurons and calretinin fibers arising from the thalamic nucleus reuniens degenerate in the subiculum after kainic acid-induced seizures. AB - The subiculum is the major output area of the hippocampus. It is closely interconnected with the entorhinal cortex and other parahippocampal areas. In animal models of temporal lobe epilepsy (TLE) and in TLE patients it exerts increased network excitability and may crucially contribute to the propagation of limbic seizures. Using immunohistochemistry and in situ-hybridization we now investigated neuropathological changes affecting parvalbumin and calretinin containing neurons in the subiculum and other parahippocampal areas after kainic acid-induced status epilepticus. We observed prominent losses in parvalbumin containing interneurons in the subiculum and entorhinal cortex, and in the principal cell layers of the pre- and parasubiculum. Degeneration of parvalbumin positive neurons was associated with significant precipitation of parvalbumin immunoreactive debris 24 h after kainic acid injection. In the subiculum the superficial portion of the pyramidal cell layer was more severely affected than its deep part. In the entorhinal cortex, the deep layers were more severely affected than the superficial ones. The decrease in number of parvalbumin positive neurons in the subiculum and entorhinal cortex correlated with the number of spontaneous seizures subsequently experienced by the rats. The loss of parvalbumin neurons thus may contribute to the development of spontaneous seizures. On the other hand, surviving parvalbumin neurons revealed markedly increased expression of parvalbumin mRNA notably in the pyramidal cell layer of the subiculum and in all layers of the entorhinal cortex. This indicates increased activity of these neurons aiming to compensate for the partial loss of this functionally important neuron population. Furthermore, calretinin-positive fibers terminating in the molecular layer of the subiculum, in sector CA1 of the hippocampus proper and in the entorhinal cortex degenerated together with their presumed perikarya in the thalamic nucleus reuniens. In addition, a significant loss of calretinin containing interneurons was observed in the subiculum. Notably, the loss in parvalbumin positive neurons in the subiculum equaled that in human TLE. It may result in marked impairment of feed-forward inhibition of the temporo-ammonic pathway and may significantly contribute to epileptogenesis. Similarly, the loss of calretinin-positive fiber tracts originating from the nucleus reuniens thalami significantly contributes to the rearrangement of neuronal circuitries in the subiculum and entorhinal cortex during epileptogenesis. PMID- 21616129 TI - Identification and localization of a neuron-specific isoform of TAF1 in rat brain: implications for neuropathology of DYT3 dystonia. AB - The neuron-specific isoform of the TAF1 gene (N-TAF1) is thought to be involved in the pathogenesis of DYT3 dystonia, which leads to progressive neurodegeneration in the striatum. To determine the expression pattern of N-TAF1 transcripts, we developed a specific monoclonal antibody against the N-TAF1 protein. Here we show that in the rat brain, N-TAF1 protein appears as a nuclear protein within subsets of neurons in multiple brain regions. Of particular interest is that in the striatum, the nuclei possessing N-TAF1 protein are largely within medium spiny neurons, and they are distributed preferentially, though not exclusively, in the striosome compartment. The compartmental preference and cell type-selective distribution of N-TAF1 protein in the striatum are strikingly similar to the patterns of neuronal loss in the striatum of DYT3 patients. Our findings suggest that the distribution of N-TAF1 protein could represent a key molecular characteristic contributing to the pattern of striatal degeneration in DYT3 dystonia. PMID- 21616130 TI - Systemic toxicity of di-2-ethylhexyl terephthalate (DEHT) in rodents following four weeks of intravenous exposure. AB - BACKGROUND: Di-2-ethylhexyl-terephtalate (DEHT) is a general purpose plasticizer and a structural isomer to di-2-ethylhexyl phthalate (DEHP) being known for its toxicity. Despite the fact that DEHT is used in quite a number of synthetics for medical device production including equipment for intravenous administration, toxicity of DEHT has not been assessed after/during intravenous exposure. Hence we report here the results of a toxicity study in male and female rats with continuous intravenous infusion of DEHT over 4 weeks. METHODS: The study was done according to OECD guidelines under GLP conditions. The dose was infused per day to male and female rats over a period of 4 weeks with saline (control), middle chain triglycerides (vehicle) as well as with 38.2, 114.5 or 381.6mg DEHT/kg. Each group (n=6) was closely monitored regarding survival, body weight development, food and water consumption. Moreover blood and urine samples were taken and a standardized necropsy as wall as a histological analysis was performed after the investigation period. RESULTS: DEHT had no effect on survival, body weight development, food and water consumption in the whole dose range investigated. There were no indications as to hematotoxicity or immunotoxicity. Clinical chemistry and histopathology indicated no exposure related effect on hepatic, thyroidal and reproductive functions or organs. CONCLUSION: DEHT administered via intravenous infusion was tolerated systemically and locally without adverse effects up to and including 381.6mg/kg/day (NOAEL=381.6mg/kg*day). In particular, there were no effects on reproductive tissues/organs, kidneys, liver hepatocytes and peroxisomes, which are known targets of DEHP-toxicity. PMID- 21616132 TI - Impact of noise or styrene exposure on the kinetics of presbycusis. AB - Presbycusis, or age-related hearing loss is a growing problem as the general population ages. In this longitudinal study, the influence of noise or styrene exposure on presbycusis was investigated in Brown Norway rats. Animals were exposed at 6 months of age, either to a band noise centered at 8 kHz at a Lex,8h = 85 dB (86.2 dB SPL for 6 h), or to 300 ppm of styrene for 6 h per day, five days per week, for four weeks. Cubic distortion product otoacoustic emissions (2f1-f2 DPOAEs) were used to test the capacity of the auditory receptor over the lifespan of the animals. 2f1-f2DPOAE measurements are easy to implement and efficiently track the age-related deterioration of mid- and high-frequencies. They are good indicators of temporary auditory threshold shift, especially with a level of primaries close to 60 dB SPL. Post-exposure hearing defects are best identified using moderate, rather than high, levels of primaries. Like many aging humans, aging rats lose sensitivity to high-frequencies faster than to medium frequencies. Although the results obtained with the styrene exposure were not entirely conclusive, histopathological data showed the presbycusis process to be enhanced. Noise-exposed rats exhibit a loss of spiral ganglion cells from 12 months and a 7 dB drop in 2f1-f2DPOAEs at 24 months, indicating that even moderate-intensity noise can accelerate the presbycusis process. Even though the results obtained with the styrene exposure are less conclusive, the histopathological data show an enhancement of the presbycusis process. PMID- 21616131 TI - Micropatterned methacrylate polymers direct spiral ganglion neurite and Schwann cell growth. AB - Significant advances in the functional outcomes achieved with cochlear implantation will likely require tissue-engineering approaches to improve the neural prosthesis interface. One strategy is to direct spiral ganglion neuron (SGN) axon growth in a highly organized fashion to approximate or contact stimulating electrodes. Here we assessed the ability of micropatterns induced by photopolymerization in methacrylate (MA) polymer systems to direct cultured neonatal rat SGN neurite growth and alignment of SG Schwann cells (SGSCs). SGN survival and neurite length were comparable among various polymer compositions. Remarkably, there was no significant difference in SGN survival or neurite length between laminin and non-laminin coated MA polymer substrates, suggesting high biocompatibility with SG tissue. Micropatterning with photopolymerization generated microchannels with a ridge periodicity of 50 MUm and channel depths of 0.6-1.0 MUm. SGN neurites grew within the grooves of the microchannels. These topographies strongly induced alignment of dissociated SGN neurites and SGSCs to parallel the pattern. By contrast, fibroblasts failed to align with the micropattern suggesting cell specific responses to topographical cues. SGN neurites extending from explants turned to parallel the pattern as they encountered the microchannels. The extent of turning was significantly correlated with angle at which the neurite initially encountered the pattern. These results indicate that SGN neurites respond to microtopographical features and that these features can be used to direct neurite growth in a highly organized fashion. PMID- 21616133 TI - Early life permethrin insecticide treatment leads to heart damage in adult rats. AB - Early life environmental exposure to xenobiotics could represent a critical period for the onset of permanent alterations in the structure and function of different organs. Cardiovascular diseases can be related to various factors including environmental toxicants. The aim of the present study was to evaluate the effect of early life permethrin treatment (1/50 LD(50), from 6th to 21st day of life) on heart of adult rats. Increased DNA damage, decreased heart cell membrane fluidity, increased cholesterol content, protein and lipid oxidation were measured in heart cells from adult rats treated with permethrin during the neonatal period with respect to control rats. Moreover, the same group showed higher levels of cholesterol, IL-1beta, IL-2, IFN-gamma, rat-Rantes and IL-10 cytokines and decreased albumin content in plasma. Lower cholesterol levels and perturbation in the phospholipid lateral diffusion together with decreased GSH levels and increased GPx activity were measured in heart mitochondria of the treated group. Our findings support the evidence that the neonatal period has a critical role in the development of heart disease in adulthood. We hypothesize that the alterations observed in adult rats could depend on epigenetic changes that occurred during this period which influence gene expression throughout the rat's life, leading to alterations of certain parameters related to cardiac function. PMID- 21616134 TI - Effects of altered glucose supply and adiposity on expression of hypothalamic energy balance regulatory genes in late gestation growth restricted ovine fetuses. AB - Intra-uterine growth restriction (IUGR) predisposes obesity in adulthood. This may be due to altered fetal nutrition causing sustained changes within the developing hypothalamic energy balance regulatory system. Using our established ovine model of IUGR, 130-day singleton fetuses (term=147 days) were obtained from growing adolescent mothers on control dietary intake (C), high intake (H) or H with growth hormone administration during either early (H+early GH) or late gestation (H+late GH) (n=6/group). GH increased maternal glycemia for the duration of treatment. H and H+early GH fetuses showed IUGR compared with C fetuses; body weight was partially restored in H+late GH fetuses, with 40% increased adiposity. In the fetal hypothalamic arcuate nucleus (ARC), cocaine- and amphetamine-regulated transcript mRNA (anorexigenic) was decreased in H fetuses and correlated across all groups with total fetal liver glycogen. Neuropeptide Y, agouti-related peptide (orexigenic) and proopiomelanocortin (anorexigenic) mRNAs were not different between groups. Insulin receptor mRNA in the ARC was increased in H, H+early GH and H+late GH fetuses and correlated negatively with fetal plasma insulin. Leptin receptor mRNA in the ARC correlated positively with fetal plasma leptin concentration and fetal fat content. Therefore, in IUGR fetuses, a key anorexigenic neuropeptide is sensitive to altered glucose supply and the hypothalamic leptin-signaling pathway is altered prenatally by increased adiposity and leptinemia. These changes could impact on postnatal energy balance regulation. PMID- 21616135 TI - Heightened locomotor-activating effects of amphetamine administered into the nucleus accumbens in adolescent rats. AB - There is a shift in sensitivity to systemically administered psychostimulants in adolescence, as evidenced by less amphetamine-induced locomotor activity in adolescent compared to adult rodents. Locomotor activating effects of amphetamine are dependent on drug actions in the core of the nucleus accumbens (NAc), but the contribution of this region to age differences in amphetamine sensitivity has not been studied directly. In the present study, we investigated the development of the NAc using targeted injections of amphetamine (0, 3, or 6 MUg/side) directly into the NAc core in early (postnatal day 30; P30) or late (P45) adolescence, or in adulthood (P75). Locomotor activity was recorded during two 1h sessions, 48 h apart. Amphetamine increased locomotor activity at all ages. P45 rats were more active than adults only at the 3 MUg/side dose, but this difference was not significant when baseline activity was taken into account. In contrast, P30 rats were more active than adults at the 6 MUg/side dose, indicating that the magnitude of the locomotor response is highest in early adolescence. Results of the present study are the first to directly show a developmental difference in the sensitivity of the NAc to amphetamine under conditions in which the influence of pharmacokinetic factors and regulatory brain regions is minimized. PMID- 21616136 TI - Origin and evolutionary analysis of the plant-specific TIFY transcription factor family. AB - A substantial number of transcription factor families have been investigated from all kingdoms of life, but a particular class of plant-specific TIFY transcription factors, characterized by a highly conserved TIFY domain, lacks a systemic analysis of its origin and evolutionary relationships among different plant species. After exhaustive genome-wide searches against 14 genomes, TIFY transcription factors were identified and classified into four subfamilies TIFY, PPD, JAZ and ZML according to their different domain architectures. Results show that the TIFY domain of the ZML subfamily possesses a core "TLS[F/Y]XG" motif rather than the "TIFYXG" motif that is dominant in the other three subfamilies. A comprehensive survey of the TIFY family allowed us to discover a new group within the JAZ subfamily and to identify several novel conserved motifs via phylogenetic analysis. Evolutional analysis indicates that whole genome duplication and tandem duplication contributed to the expansion of the TIFY family in plants. PMID- 21616137 TI - Distinct role of Hsp70 in Drosophila hemocytes during severe hypoxia. AB - Severe hypoxia can lead to injury and mortality in vertebrate or invertebrate organisms. Our research is focused on understanding the molecular mechanisms that lead to injury or adaptation to hypoxic stress using Drosophila as a model system. In this study, we employed the UAS-Gal4 system to dissect the protective role of Hsp70 in specific tissues in vivo under severe hypoxia. In contrast to overexpression in tissues such as muscles, heart, and brain, we found that overexpression of Hsp70 in hemocytes of flies provides a remarkable survival benefit to flies exposed to severe hypoxia for days. Furthermore, these flies were tolerant not only to severe hypoxia but also to other stresses such as oxidant stress (e.g., paraquat feeding or hyperoxia). Interestingly we observed that the better survival with Hsp70 overexpression in hemocytes under hypoxia or oxidant stress is causally linked to reactive oxygen species (ROS) reduction in whole flies. We also show that hemocytes are a major source of ROS generation, leading to injury during hypoxia, and their elimination results in a better survival under hypoxia. Hence, our study identified a protective role for Hsp70 in Drosophila hemocytes, which is linked to ROS reduction in the whole flies and thus helps in their remarkable survival during oxidant or hypoxic stress. PMID- 21616139 TI - P66Shc mediated ferritin degradation--a novel mechanism of ROS formation. AB - Diallyl trisulfide (DATS) has been shown to induce the formation of reactive oxygen species (ROS) in prostate cancer cells, which was accompanied by a decrease in the ferritin protein level and an increase in the labile iron pool (LIP). However, the mechanism of the ferritin degradation has not been fully elucidated. In this paper we demonstrate that DATS-induced ROS formation depends on p66Shc. In cells stably expressing a dominant negative mutant of p66Shc (p66ShcS36A), DATS did not induce ROS formation. In addition, in cells expressing p66ShcS36A neither an increase in ferritin H degradation nor an increase in LIP were observed. Cells stably expressing p66ShcS36A also possess higher levels of ferritin H compared to PC-3 cells transfected with an empty vector. Moreover, DATS-induced G2/M arrest is completely abrogated in cells expressing p66ShcS36A. Mouse embryonic fibroblasts (MEFs) derived from wild-type (WT) or p66Shc knockout mouse have been used to evaluate if p66Shc involvement in DATS-induced signaling is cell specific. DATS induced G2/M arrest in WT MEFs but had no effect in the p66Shc(-/-) cell line. Moreover, increases in LIP and ROS formation were significantly attenuated in p66Shc(-/-) MEFs treated with DATS. PMID- 21616140 TI - Salubrinal, an eIF2alpha dephosphorylation inhibitor, enhances cisplatin-induced oxidative stress and nephrotoxicity in a mouse model. AB - Although cisplatin attacks various tumors with remarkable efficacy, its clinical usage has been limited by its side effects, particularly nephrotoxicity. Salubrinal, a selective eukaryotic translation initiation factor 2 subunit alpha (eIF2alpha) dephosphorylation inhibitor, has been found to protect cells from endoplasmic reticulum (ER)-stress-induced cytotoxicity. In this study, we hypothesized that salubrinal would protect against cisplatin-induced nephrotoxicity in a mouse model. Cisplatin treatment significantly increased serum blood urea nitrogen and creatinine levels, renal kidney injury marker (kim 1) mRNA expression, renal cell apoptosis, and renal histopathological changes in mice. Unexpectedly, administration of salubrinal significantly enhanced the cisplatin-induced nephrotoxicity in mice. Salubrinal by itself did not induce alterations in the function or histomorphology of mouse kidneys. Salubrinal significantly enhanced the phosphorylation of eIF2alpha, the protein expression of activating transcription factor 4 and CCAAT/enhancer binding protein homologous protein, and the cleavage of caspases 12, 9, and 3 in the kidneys of cisplatin-treated mice. Moreover, salubrinal enhanced the cisplatin-induced oxidative stress in the kidneys. The antioxidant N-acetylcysteine significantly reversed the increased renal lipid peroxidation, activated renal caspase cascade, and increased blood BUN and creatinine in cisplatin-alone- or cisplatin plus salubrinal-treated mice. These findings suggest that salubrinal aggravates cisplatin-induced nephrotoxicity through the enhancement of oxidative stress and ER stress-related cell apoptosis. PMID- 21616138 TI - Regulation of Ras proteins by reactive nitrogen species. AB - Ras GTPases have been a subject of intense investigation since the early 1980s, when single point mutations in Ras were shown to cause deregulated cell growth control. Subsequently, Ras was identified as the most prevalent oncogene found in human cancer. Ras proteins regulate a host of pathways involved in cell growth, differentiation, and apoptosis by cycling between inactive GDP-bound and active GTP-bound states. Regulation of Ras activity is controlled by cellular factors that alter guanine nucleotide cycling. Oncogenic mutations prevent protein regulatory factors from down-regulating Ras activity, thereby maintaining Ras in a chronically activated state. The central dogma in the field is that protein modulatory factors are the primary regulators of Ras activity. Since the mid 1990s, however, evidence has accumulated that small molecule reactive nitrogen species (RNS) can also influence Ras guanine nucleotide cycling. Herein, we review the basic chemistry behind RNS formation and discuss the mechanism through which various RNS enhance nucleotide exchange in Ras proteins. In addition, we present studies that demonstrate the physiological relevance of RNS-mediated Ras activation within the context of immune system function, brain function, and cancer development. We also highlight future directions and experimental methods that may enhance our ability to detect RNS-mediated activation in cell cultures and in vivo. The development of such methods may ultimately pave new directions for detecting and elucidating how Ras proteins are regulated by redox species, as well as for targeting redox-activated Ras in cancer and other disease states. PMID- 21616141 TI - Ironing out neurodegeneration: iron chelation for neuroprotection. PMID- 21616144 TI - The emerging role of HSP20 as a multifunctional protective agent. AB - The small heat shock proteins (sHSPs) are a highly conserved family of molecular chaperones that are ubiquitously expressed throughout nature. They are transiently upregulated in many tissue types following stressful stimuli. Recently, one member of the sHSP family, HSP20 (HspB6), has been shown to be highly effective as a protective mediator against a number of debilitating pathological conditions, including cardiac hypertrophy and Alzheimer's disease. Hsp20 is also an important modulator of vital physiological processes, such as smooth muscle relaxation and cardiac contractility. This review focuses on the molecular mechanisms employed by HSP20 that allow it to act as an innate protector in the context of cardiovascular and neurological diseases. Emerging evidence for a possible role as an anti-cancer agent is also presented. PMID- 21616143 TI - Mitochondrial stress: a bridge between mitochondrial dysfunction and metabolic diseases? AB - Under pathophysiological conditions such as obesity, excessive oxidation of nutrients may induce mitochondrial stress, leading to mitochondrial unfolded protein response (UPR(mt)) and initiation of a retrograde stress signaling pathway. Defects in the UPR(mt) and the retrograde signaling pathways may disrupt the integrity and homeostasis of the mitochondria, resulting in endoplasmic reticulum stress and insulin resistance. Improving the capacity of mitochondria to reduce stress may be an effective approach to improve mitochondria function and to suppress obesity-induced metabolic disorders such as insulin resistance and type 2 diabetes. PMID- 21616142 TI - Diverse functions of cationic Mn(III) N-substituted pyridylporphyrins, recognized as SOD mimics. AB - Oxidative stress, a redox imbalance between the endogenous reactive species and antioxidant systems, is common to numerous pathological conditions such as cancer, central nervous system injuries, radiation injury, diabetes etc. Therefore, compounds able to reduce oxidative stress have been actively sought for over 3 decades. Superoxide is the major species involved in oxidative stress either in its own right or through its progeny, such as ONOO-, H2O2, *OH, CO3*-, and *NO2. Hence, the very first compounds developed in the late 1970-ies were the superoxide dismutase (SOD) mimics. Thus far the most potent mimics have been the cationic meso Mn(III) N-substituted pyridylporphyrins and N,N'-disubstituted imidazolylporphyrins (MnPs), some of them with k(cat)(O2.-) similar to the k(cat) of SOD enzymes. Most frequently studied are ortho isomers MnTE-2-PyP5+, MnTnHex-2 PyP5+, and MnTDE-2-ImP5+. The ability to disproportionate O2.- parallels their ability to remove the other major oxidizing species, peroxynitrite, ONOO-. The same structural feature that gives rise to the high k(cat)(O2.-) and k(red)(ONOO ), allows MnPs to strongly impact the activation of the redox-sensitive transcription factors, HIF-1alpha, NF-kappaB, AP-1, and SP-1, and therefore modify the excessive inflammatory and immune responses. Coupling with cellular reductants and other redox-active endogenous proteins seems to be involved in the actions of Mn porphyrins. While hydrophilic analogues, such as MnTE-2-PyP5+ and MnTDE-2-ImP5+ are potent in numerous animal models of diseases, the lipophilic analogues, such as MnTnHex-2-PyP5+, were developed to cross blood brain barrier and target central nervous system and critical cellular compartments, mitochondria. The modification of its structure, aimed to preserve the SOD-like potency and lipophilicity, and diminish the toxicity, has presently been pursued. The pulmonary radioprotection by MnTnHex-2-PyP5+ was the first efficacy study performed successfully with non-human primates. The Phase I toxicity clinical trials were done on amyotrophic lateral sclerosis patients with N,N' diethylimidazolium analogue, MnTDE-2-ImP5+ (AEOL10150). Its aggressive development as a wide spectrum radioprotector by Aeolus Pharmaceuticals has been supported by USA Federal government. The latest generation of compounds, bearing oxygens in pyridyl substituents is presently under aggressive development for cancer and CNS injuries at Duke University and is supported by Duke Translational Research Institute, The Wallace H. Coulter Translational Partners Grant Program, Preston Robert Tisch Brain Tumor Center at Duke, and National Institute of Allergy and Infectious Diseases. Metal center of cationic MnPs easily accepts and donates electrons as exemplified in the catalysis of O2.- dismutation. Thus such compounds may be equally good anti- and pro-oxidants; in either case the beneficial therapeutic effects may be observed. Moreover, while the in vivo effects may appear antioxidative, the mechanism of action of MnPs that produced such effects may be pro-oxidative; the most obvious example being the inhibition of NF-kappaB. The experimental data therefore teach us that we need to distinguish between the mechanism/s of action/s of MnPs and the effects we observe. A number of factors impact the type of action of MnPs leading to favorable therapeutic effects: levels of reactive species and oxygen, levels of endogenous antioxidants (enzymes and low-molecular compounds), levels of MnPs, their site of accumulation, and the mutual encounters of all of those species. The complexity of in vivo redox systems and the complex redox chemistry of MnPs challenge and motivate us to further our understanding of the physiology of the normal and diseased cell with ultimate goal to successfully treat human diseases. PMID- 21616145 TI - Function of the asparagine 74 residue of the inhibitory gamma-subunit of retinal rod cGMP-phophodiesterase (PDE) in vivo. AB - The inhibitory subunit of rod cyclic guanosine monophosphate (cGMP) phosphodiesterase, PDE6gamma, is a major component of rod transduction and is required to support photoreceptor integrity. The N74A allele of PDE6gamma has previously been shown in experiments carried out in vitro to reduce the regulatory inhibition on the PDE6 catalytic core subunits, PDE6alphabeta. This should, in intact rods, lead to an increase in basal (dark) PDE6 activity producing a state equivalent to light adaptation in the rods and we have examined this possibility using ERG and suction-electrode measurements. The murine opsin promoter was used to drive the expression of a mutant N74A and a wild-type PDE6gamma control transgene in the photoreceptors of +/Pde6g(tm1) mice. This transgenic line was crossed with Pde6g(tm1)/Pde6g(tm1) mice to generate animals able to synthesize only the transgenic mutant PDE6gamma. We find that the N74A mutation did not produce a significant decrease in circulating current, a decrease in sensitivity or affect the kinetics of the light response, all hallmarks of the light-adapted state. In an in vitro assay of the PDE purified from the N74A transgenic mice and control mice we could find no increase in basal activity of the mutant PDE6. Both the results from the physiology and the biochemistry experiments are consistent with the interpretation that the mutation causes a much milder phenotype in vivo than was predicted from observations made using a cell-free assay system. The in vivo regulation of PDE6gamma on PDE6alphabeta may be more dynamic and context-dependent than was replicated in vitro. PMID- 21616146 TI - Interaction of the receptor FGFRL1 with the negative regulator Spred1. AB - FGFRL1 is a member of the fibroblast growth factor receptor family. It plays an essential role during branching morphogenesis of the metanephric kidneys, as mice with a targeted deletion of the Fgfrl1 gene show severe kidney dysplasia. Here we used the yeast two-hybrid system to demonstrate that FGFRL1 binds with its C terminal, histidine-rich domain to Spred1 and to other proteins of the Sprouty/Spred family. Members of this family are known to act as negative regulators of the Ras/Raf/Erk signaling pathway. Truncation experiments further showed that FGFRL1 interacts with the SPR domain of Spred1, a domain that is shared by all members of the Sprouty/Spred family. The interaction could be verified by coprecipitation of the interaction partners from solution and by codistribution at the cell membrane of COS1 and HEK293 cells. Interestingly, Spred1 increased the retention time of FGFRL1 at the plasma membrane where the receptor might interact with ligands. FGFRL1 and members of the Sprouty/Spred family belong to the FGF synexpression group, which also includes FGF3, FGF8, Sef and Isthmin. It is conceivable that FGFRL1, Sef and some Sprouty/Spred proteins work in concert to control growth factor signaling during branching morphogenesis of the kidneys and other organs. PMID- 21616147 TI - Fat-1 transgenic mice with elevated omega-3 fatty acids are protected from allergic airway responses. AB - Omega-3 polyunsaturated fatty acids (n-3 PUFA) have been implicated in the alleviation of asthma. Recent studies have demonstrated that the n-3 PUFA derived lipid mediators, protectin D1 and resolvin E1, may act as potent resolution agonists in airway inflammation. The effects of the n-3 PUFA tissue status itself on asthma pathogenesis remains to be further investigated. In this study allergic airway inflammation induced by allergen sensitization and aerosol challenge in Fat-1 and wild-type mice was investigated. Fat-1 transgenic mice displayed increased endogenous lung n-3 PUFA. When allergen-sensitized and aerosol challenged, these animals had decreased airway inflammation with decreased leukocyte accumulation in bronchoalveolar lavage fluid and lung parenchyma. The Fat-1 mice had a shift to the right in the dose-response relationship for methacholine induced bronchoconstriction with a significant increase in the log ED200. The Fat-1 mice had lower BALF concentrations of the pro-inflammatory cytokines IL-1alpha, IL-2, IL-5, IL-9, IL-13, G-CSF, KC and RANTES. Furthermore, increased lung tissue amounts of the counter-regulatory mediators protectin D1 and resolvin E1 were found in Fat-1 mice after bronchoprovocative challenge. These results therefore demonstrate a direct protective role for lung n-3 PUFA in allergic airway responses and an increased generation of protectin D1 and resolvin E1 in this context. PMID- 21616148 TI - Bioactivity and toxicity studies of amphotericin B incorporated in liquid crystals. AB - The main objective of the present work was to evaluate the bioactivity and safety of amphotericin B-liquid crystal mixtures (AmB-LC). The effects of liquid crystal (LC) materials and AmB-LC ratios on bioactivity and toxicity to respiratory cell lines were investigated. The formation of AmB-LC mixtures did not change the physical properties of LC when the AmB loading was not more than a 1:3 mole ratio (25%). The exposure of respiratory cell lines to LC did not generate any toxicity (2.5-80 MUg/mL). The inhibitory activity of AmB in all liquid crystal formulations (cholesteryl palmityl carbonate: CPC, dicholesteryl carbonate: DCC and sodium cholesteryl carbonate: SCC) on fungi was significantly enhanced when compared to that of the same amount of pure AmB. However, their toxicity to respiratory related cells and red blood cells was significantly decreased. This could be a huge advantage in clinical applications as there is more possibility for dose adjustments. The exposure of small airway epithelial cells (SAEC) and alveolar macrophages (AMs NR8383) to liquid crystals had no significant detrimental effects at doses of between 2.5 and 80 MUg/mL (viability was always over 80%). The production of toxic cytokines and inflammatory mediators, including tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta) and nitric oxide (NO), after treatment with AmB in liquid crystals at concentrations of between 2 and 32 MUg/mL was significantly reduced by about a 1000-fold compared to that generated by lipopolysaccharide (LPS). PMID- 21616149 TI - Decrease of blood type antigenicity over the long-term after ABO-incompatible kidney transplantation. AB - BACKGROUND: Few studies have investigated the changes in the antigenicities of the transplanted organs after transplantation. METHODS: We examined, by immunohistochemical assay, the changes in expression of the blood-type antigens on the transplanted kidneys over the long-term after ABO-incompatible kidney transplantation with A- or B-antigen incompatibility (blood type A to B and B to A). The subjects were six patients, including one case with graft loss, who had received ABO-incompatible kidney allografts more than ten years previously. As normal controls, four cases of ABO-compatible transplantation during the same period, including two recipient/donor pairs each with blood group A1 and blood group B were enrolled. RESULTS: Expression of the blood-type A or B antigens decreased gradually to 91.8% during the first three months after transplantation, 85.8% during the first five years, 64.1% during the first ten years, and 57.6% over ten years after transplantation on average in ABO-incompatible transplant recipients. In one patient with graft loss due to severe antibody-mediated rejection, the donor's type B blood-type antigens on the transplanted graft had changed but partially to the recipient's blood-type A antigen by 2582days after the transplantation, suggestive of the occurrence of blood-type chimerism on the endothelium. In ABO-compatible transplant recipients, such changes in expression were not observed. The average percentage of blood-type antigen-positive vessels at more than ten years after the renal transplantation was 99.8%. CONCLUSIONS: Decrease in the expression of the donor's blood-type antigen on the endothelium of the graft has been considered as one of the mechanisms underlying the accommodation occurring over the long-term after ABO-incompatible kidney transplantation. On the other hand, establishment of antigenic chimerism on the graft endothelium could be one of the hallmarks of the immunological reaction associated with antibody-mediated rejection. PMID- 21616150 TI - System approaches of Weiss and Bertalanffy and their relevance for systems biology today. AB - System approaches in biology have a long history. We focus here on the thinking of Paul A. Weiss and Ludwig von Bertalanffy, who contributed a great deal towards making the system concept operable in biology in the early 20th century. To them, considering whole living systems, which includes their organisation or order, is equally important as the dynamics within systems and the interplay between different levels from molecules over cells to organisms. They also called for taking the intrinsic activity of living systems and the conservation of system states into account. We compare these notions with today's systems biology, which is often a bottom-up approach from molecular dynamics to cellular behaviour. We conclude that bringing together the early heuristics with recent formalisms and novel experimental set-ups can lead to fruitful results and understanding. PMID- 21616151 TI - Improved expression of Rhizopus oryzae alpha-amylase in the methylotrophic yeast Pichia pastoris. AB - In our previous study, the alpha-amylase from Rhizopus oryzae (RoAmy) was expressed in Escherichia coli and Saccharomyces cerevisiae but the obtained recombinant RoAmy (rRoAmy) yields were too low. The aim of the present research was to obtain high-level expressions of RoAmy in the methylotrophic yeast Pichia pastoris. To this end, we constructed P. pastoris strains with the capability to express recombinant alpha-amylase under the control of constitutive glyceraldehyde-3-phosphate dehydrogenase (GAP) and methanol-inducible alcohol oxidase 1 promoters. The levels of inducibly expressed rRoAmy were higher than those of constitutively expressed. The maximal inducible rRoAmy expression levels for the Mut(+) strains (41.1mg/l) were approximately eight times higher than those for the Mut(s) strains and 24 times higher than those expressed under the control of the GAP promoter. For both inducible and constitutive expressions, the S. cerevisiae alpha-prepro sequence and the native signal sequence of RoAmy were used separately to direct the secretion of rRoAmy into the culture medium of P. pastoris. Low levels of intracellular amylase activities that had been detected after shake-flask fermentation indicated that both signal sequences could effectively direct the secretion of rRoAmy under all studied conditions. In addition, the secretion levels of rRoAmy directed with its own signal peptide were 7-10% higher than those directed by the alpha-prepro sequence. PMID- 21616152 TI - A differential structure approach to membrane segmentation in electron tomography. AB - Electron tomography allows three-dimensional visualization of cellular landscapes in molecular detail. Segmentation is a paramount stage for the interpretation of the reconstructed tomograms. Although several computational approaches have been proposed, none has prevailed as a generic method and thus segmentation through manual annotation is still a common choice. In this work we introduce a segmentation method targeted at membranes, which define the natural limits of compartments within biological specimens. Our method is based on local differential structure and on a Gaussian-like membrane model. First, it isolates information through scale-space and finds potential membrane-like points at a local scale. Then, the structural information is integrated at a global scale to yield the definite segmentation. We show and validate the performance of the algorithm on a number of tomograms under different experimental conditions. PMID- 21616153 TI - Clustering and variance maps for cryo-electron tomography using wedge-masked differences. AB - Cryo-electron tomography provides 3D imaging of frozen hydrated biological samples with nanometer resolution. Reconstructed volumes suffer from low signal to-noise-ratio (SNR)(1) and artifacts caused by systematically missing tomographic data. Both problems can be overcome by combining multiple subvolumes with varying orientations, assuming they contain identical structures. Clustering (unsupervised classification) is required to ensure or verify population homogeneity, but this process is complicated by the problems of poor SNR and missing data, the factors that led to consideration of multiple subvolumes in the first place. Here, we describe a new approach to clustering and variance mapping in the face of these difficulties. The combined subvolume is taken as an estimate of the true subvolume, and the effect of missing data is computed for individual subvolumes. Clustering and variance mapping then proceed based on differences between expected and observed subvolumes. We show that this new method is faster and more accurate than two current, widely used techniques. PMID- 21616154 TI - Ventral striatal signal changes represent missed opportunities and predict future choice. AB - Realizing one has missed an opportunity can influence decision behavior in the future, such that a large missed opportunity leads to more risk taking in the next round. To investigate the neuronal mechanism of this phenomenon we used functional magnetic resonance imaging (fMRI) in combination with a sequential decision task in which the magnitude of possible gains linearly increased, but at the same time the gain probability decreased. After subjects decided to stop a trial and to collect the gains, not only the chosen option (actual outcome), but also the alternative option (maximum possible gain in this round) was revealed. Our data show that a missed chance influenced volunteers' decision behavior: volunteers took more risk after rounds in which they had missed a large opportunity. This was paralleled by signal changes in a lateral area of the ventral striatum that scaled with the difference between what could have been gained and what was actually gained in this round. In addition, after gains signal changes in dopaminoceptive structures including the midbrain and ventral striatum together with the insula predicted individual choice behavior in the subsequent round. Thus, our data provide a neural mechanism for how missed opportunities influence future decisions. PMID- 21616155 TI - Inhibition in early Alzheimer's disease: an fMRI-based study of effective connectivity. AB - Changes of functional connectivity in prodromal and early Alzheimer's disease can arise from compensatory and/or pathological processes. We hypothesized that i) there is impairment of effective inhibition associated with early Alzheimer's disease that may lead to ii) a paradoxical increase of functional connectivity. To this end we analyzed effective connectivity in 14 patients and 16 matched controls using dynamic causal modeling of functional MRI time series recorded during a visual inter-hemispheric integration task. By contrasting co-linear with non co-linear bilateral gratings, we estimated inhibitory top-down effects within the visual areas. The anatomical areas constituting the functional network of interest were identified with categorical functional MRI contrasts (Stimuli>Baseline and Co-linear gratings>Non co-linear gratings), which implicated V1 and V3v in both hemispheres. A model with reciprocal excitatory intrinsic connections linking these four regions and modulatory inhibitory effects exerted by V3v on V1 optimally explained the functional MRI time series in both subject groups. However, Alzheimer's disease was associated with significantly weakened intrinsic and modulatory connections. Top-down inhibitory effects, previously detected as relative deactivations of V1 in young adults, were observed neither in our aged controls nor in patients. We conclude that effective inhibition weakens with age and more so in early Alzheimer's disease. PMID- 21616156 TI - Ventromedial prefrontal volume predicts understanding of others and social network size. AB - Cognitive abilities such as Theory of Mind (ToM), and more generally mentalizing competences, are central to human sociality. Neuroimaging has associated these abilities with specific brain regions including temporo-parietal junction, superior temporal sulcus, frontal pole, and ventromedial prefrontal cortex. Previous studies have shown both that mentalizing competence, indexed as the ability to correctly understand others' belief states, is associated with social network size and that social group size is correlated with frontal lobe volume across primate species (the social brain hypothesis). Given this, we predicted that both mentalizing competences and the number of social relationships a person can maintain simultaneously will be a function of gray matter volume in these regions associated with conventional Theory of Mind. We used voxel-based morphometry of Magnetic Resonance Images (MRIs) to test this hypothesis in humans. Specifically, we regressed individuals' mentalizing competences and social network sizes against gray matter volume. This revealed that gray matter volume in bilateral posterior frontal pole and left temporoparietal junction and superior temporal sucus varies parametrically with mentalizing competence. Furthermore, gray matter volume in the medial orbitofrontal cortex and the ventral portion of medial frontal gyrus, varied parametrically with both mentalizing competence and social network size, demonstrating a shared neural basis for these very different facets of sociality. These findings provide the first fine-grained anatomical support for the social brain hypothesis. As such, they have important implications for our understanding of the constraints limiting social cognition and social network size in humans, as well as for our understanding of how such abilities evolved across primates. PMID- 21616157 TI - Keeping the rhythm -- pro-arrhythmic investigations in isolated Gottingen minipig hearts. AB - INTRODUCTION: Cardiac arrhythmia is a potentially lethal condition. A better prediction and mechanistic understanding of human cardiac arrhythmia is dependent on the development of good animal models. Minipigs are increasingly being used in cardiovascular research and we hypothesize that Langendorff-perfused minipig hearts could serve as a novel model of pro-arrhythmia. METHODS: We studied arrhythmic biomarkers and overt arrhythmia using sharp electrode recordings and Langendorff-perfusion of minipig and dog hearts. Hearts were subjected to pharmacological blockade of repolarizing currents, chemical and surgical AV block, and programmed electrical stimulation. RESULTS: Minipigs have significantly longer cardiac action potentials and QT intervals than dogs at comparable cycle lengths. In addition, minipigs were relatively resistant to arrhythmias in a variety of settings and arrhythmias were only observed in response to programmed electrical S1-S2 stimulation after surgical AV-ablation and combined I(Kr) and I(Ks) blockade. Accordingly, the short term variability of the action potential remained very low in minipigs. In addition, the electro mechanical window (EMw) remained positive in both dog and minipig experiments, although the EMw approached negative values in dogs during I(Ks) blockade and beta-adrenergic stimulation. Isoprenaline- or pacing-induced changes in frequency resulted in paradoxical transient effects on repolarization. DISCUSSION: The evidence presented indicates that Langendorff-perfused Gottingen minipig hearts are resistant to the development of arrhythmias in experimental settings known to be pro-arrhythmic in other species including dogs. A possible mechanistic explanation could be the long electrical systole compared to dogs, combined with a short vulnerable window, recorded in minipig hearts. We also find that EMw recordings from Langendorff perfused hearts are less sensitive to I(Ks) blockade than what has been reported from in vivo experiments, most likely due to the mechanical properties of the recording apparatus. PMID- 21616158 TI - Deformational behaviour of knee cartilage and changes in serum cartilage oligomeric matrix protein (COMP) after running and drop landing. AB - OBJECTIVE: To investigate (1) the effect of running and drop landing interventions on knee cartilage deformation and serum cartilage oligomeric matrix protein (COMP) concentration and (2) if the changes in cartilage volume correlate with the changes in serum COMP level. METHODS: Knee joint cartilage volume and thickness were determined using magnetic resonance imaging (MRI) as well as COMP concentration from serum samples before and after in vivo loading of 14 healthy adults (seven male and seven female). Participants performed different loading interventions of 30 min duration on three different days: (1) 100 vertical drop landings from a 73 cm high platform, (2) running at a velocity of 2.2m/s (3.96 km), and (3) resting on a chair. Blood samples were taken immediately before, immediately after and 0.5h, 1h, 2h and 3h post intervention. Pre- and post loading coronal and axial gradient echo MR images with fat suppression were used to determine the patellar, tibial and femoral cartilage deformation. RESULTS: Serum COMP levels increased immediately after the running (+30.7%, pre: 7.3U/l, 95% confidence interval (CI): 5.6, 8.9, post: 9.1U/l, 95% CI: 7.2, 11.0, P=0.001) and after drop landing intervention (+32.3%, pre: 6.8U/l, 95% CI: 5.3, 8.4; post: 8.9U/l, 95% CI: 6.8, 10.9, P=0.001). Cartilage deformation was more pronounced after running compared to drop landing intervention, with being significant (volume: P=0.002 and thickness: P=0.001) only in the lateral tibia. We found a significant correlation (r(2)=0.599, P=0.001) between changes in serum COMP (%) and in cartilage volume (%) after the drop landing intervention, but not after running. CONCLUSIONS: In vivo exercise interventions differentially regulate serum COMP concentrations and knee cartilage deformations. The relation between changes in COMP and in cartilage volume seems to depend on both mechanical and biochemical factors. PMID- 21616159 TI - Short- and long-term, salinity-induced modulation of V-ATPase activity in the posterior gills of the true freshwater crab, Dilocarcinus pagei (Brachyura, Trichodactylidae). AB - To better understand the biochemical mechanisms underlying anisosmotic extracellular regulation in the freshwater Brachyura, we kinetically characterized the V-ATPase from the posterior gills of Dilocarcinus pagei, acclimated for 10days to salinities up to 210/00. Specific activity was highest in fresh water (26.5+/-2.1U mg(-1)), decreasing in 50/00 to 210/00, attaining 3 fold less at 150/00. Apparent affinities for ATP and Mg(2+) respectively increased 3.2- and 2-fold at 100/00, suggesting expression of different isoenzymes. In a 240-h time-course study of exposure to 210/00, maximum specific activity decreased 2.5- to 4-fold within 1 to 24h while apparent affinities for ATP and Mg(2+) respectively increased by 12-fold within 24h and 2.4-fold after 1h, unchanged thereafter. K(I) for bafilomycin A(1) decreased 150-fold after 1h, remaining constant up to 120h. This is the first kinetic analysis of V-ATPase specific activity in crustacean gills during salinity acclimation. Our findings indicate active gill Cl(-) uptake by D. pagei in fresh water, and short- and long term down-regulation of V-ATPase-driven ion uptake processes during salinity exposure, aiding in comprehension of the biochemical adaptations underpinning the establishment of the Brachyura in fresh water. PMID- 21616160 TI - Regulation of hexokinase by reversible phosphorylation in skeletal muscle of a freeze-tolerant frog. AB - Hexokinase (HK) was isolated from hind leg skeletal muscle of the wood frog, Rana sylvatica, a freeze tolerant species that uses glucose as a cryoprotectant. Analysis of kinetic parameters (K(m) and V(max)) of HK showed significant increases in K(m) glucose (from 144 +/- 4.4 to 248 +/- 1 2.0 MUM) and K(m) ATP (from 248 +/- 8.5 to 330 +/- 20.9 MUM), as well as a decrease in V(max) (from 86.1 +/- 0.40 to 52 +/- 0.49 mUmg(-1) of protein) in frogs following freezing exposure, indicating lower affinity for HK substrates and lower enzyme activity in this state. Subsequent analyses indicated that differential phosphorylation of HK between the two states was responsible for the altered kinetic properties. HK was analyzed by SDS-PAGE; phosphoprotein staining revealed a 33% decrease in phosphate content of HK from frozen frogs but immunoblotting showed no change in total HK protein content. Muscle extracts from control and frozen frogs were incubated with ions and second messengers to stimulate the actions of protein kinases and protein phosphatases, with results indicating that HK can be phosphorylated by protein kinases A and C, and AMP-activated protein kinase, and can be dephosphorylated by protein phosphatases 1, 2A and 2C. The data indicate that in control frogs, HK is in a higher phosphate form and displays a high substrate affinity and high activity, whereas in frozen frogs HK is less phosphorylated, with lower substrate affinity and lower activity. Studies also showed that HK affinity for ATP decreases further in response to low temperature, but that high cryoprotective glucose concentrations can prevent these changes in affinity. Finally, the activity and structure of HK from frozen frogs is more sensitive to non-compatible osmolytes than the enzyme in control frogs. PMID- 21616161 TI - [Isolation of Beauveria bassiana in a bronchoalveolar lavage specimen from a patient with bladder cancer]. PMID- 21616162 TI - PEXEL-independent trafficking of Plasmodium falciparum SURFIN4.2 to the parasite infected red blood cell and Maurer's clefts. AB - SURFIN(4.2) is a parasite-infected red blood cell (iRBC) surface associated protein of Plasmodium falciparum. To analyze the region responsible for the intracellular trafficking of SURFIN(4.2) to the iRBC and Maurer's clefts, a panel of transgenic parasite lines expressing recombinant SURFIN(4.2) fused with green fluorescent protein was generated and evaluated for their localization. We found that the cytoplasmic region containing a tryptophan rich (WR) domain is not necessary for trafficking, whereas the transmembrane (TM) region was. Two PEXEL like sequences were shown not to be responsible for the trafficking of SURFIN(4.2), demonstrating that the protein is trafficked in a PEXEL-independent manner. N-terminal replacement, deletion of the cysteine-rich domain or the variable region also did not prevent the protein from localizing at the iRBC or Maurer's clefts. A recombinant SURFIN(4.2) protein possessing 50 amino acids upstream of the TM region, TM region itself and a part of the cytoplasmic region was shown to be trafficked into the iRBC and Maurer's clefts, suggesting that there are no essential trafficking motifs in the SURFIN(4.2) extracellular region. A mini-SURFIN(4.2) protein containing WR domain was shown by Western blotting to be more abundantly detected in a Triton X-100-insoluble fraction, compared to the one without WR domain. We suggest that the cytoplasmic region containing the WR may be responsible for their difference in solubility. PMID- 21616163 TI - Cardicola opisthorchis n. sp. (Trematoda: Aporocotylidae) from the Pacific bluefin tuna, Thunnus orientalis (Temminck & Schlegel, 1844), cultured in Japan. AB - A new aporocotylid blood fluke is described, based on specimens from the ventricle of the Pacific bluefin tuna, Thunnus orientalis (Temminck et Schlegel), cultured in Wakayama and Nagasaki Prefectures, Japan. The new species is morphologically similar to the members of the genus Cardicola Short, 1953, but shows distinct differences in the body form, location of the testis and the orientation of the ootype. The body of the new species is long and slender, whereas other Cardicola species are small and generally lanceolate. The testis is mostly located posterior to the caeca and anterior to the ovary, occupying 31-45% of body length, in contrast to the known Cardicola species, whose testis is typically intercaecal. The ootype is oriented anteriorly, while in most congeners, it is directed posteriorly or horizontally. Phylogenetic analyses of this aporocotylid, together with Cardicola orientalis Ogawa, Tanaka, Sugihara et Takami, 2010 from the same host, were conducted based on DNA sequences of the ITS2 rDNA and the 28S region of ribosomal RNA. The analyses revealed that the new blood fluke belongs to the genus Cardicola despite the marked morphological differences. Thus, this aporocotylid is named Cardicola opisthorchis n. sp. and the generic diagnosis is emended in this paper. In addition, 100% identity among the ITS2 sequences from the present species, Cardicola sp. from T. orientalis in Mexico and Cardicola sp. from the northern bluefin tuna, Thunnus thynnus (Linnaeus) in Spain suggests that C. opisthorchis n. sp. has a broad geographical distribution and that it infects both the Pacific and northern bluefin tuna. PMID- 21616164 TI - Responses of thioredoxin 1 and thioredoxin-related protein 14 mRNAs to cadmium and copper stresses in Venerupis philippinarum. AB - Thioredoxin (abbreviated as Trx) is an important ubiquitous disulfide reductase, which can protect organisms against various oxidative stresses. In the present study, thioredoxin 1 (named as VpTrx1) and thioredoxin-related protein (named as VpTrp14) were identified from Venerupis philippinarum, respectively. Similar to most Trx1s, VpTrx1 possessed all conserved features critical for the fundamental structure and function of Trx1s, such as the conserved catalytic residues (C-G-P C), but lacked the other cysteine residues, while VpTrp14 contained the conserved motif (C-P-D-C). Quantitative Real-time PCR assay showed that VpTrx1 and VpTrp14 transcripts were distributed in a wide array of tissues most abundantly expressed in the hepatopancreas. The expression of VpTrp14 mRNA in the hepatopancreas was significantly up-regulated after exposure to 10 and 40MUg/L Cd, while the VpTrx1 expression level was kept relatively constant. Both the expression levels of VpTrx1 and VpTrp14 in the hepatopancreas were induced after exposure to Cu, and increased to the peak value at 96h under the 40MUg/L Cu exposure. These results showed that VpTrp14 transcripts responded to metal stress more acutely than VpTrx1, and both Trxs responded to Cu stress more sensitively than Cd. Together, it was suggested that VpTrx1 and VpTrp14 perhaps played important roles in the antioxidant responses against metal stress in V. philippinarum. PMID- 21616166 TI - Oxidative stress in newly-hatched Chorthippus brunneus--the effects of zinc treatment during diapause, depending on the female's age and its origins. AB - The responses of glutathione, glutathione S-transferases (GSTs), and catalase (CAT) were determined in 1-day-old larvae of Chorthippus brunneus Thunberg, 1815, a grasshopper exposed to zinc during diapause, from unpolluted (Pilica) or polluted (Olkusz, Szopienice) sites. The aim of the work was to search for differences among populations of the insects as a result of various multistress pressures in their habitats. The question of zinc toxicity in the context of energy allocation was also considered. Zinc caused a decrease in glutathione concentration in the body of zinc-treated larvae. Significant differences between control and zinc-treated groups were confirmed for young females' progeny from Pilica and Olkusz as well as old females' progeny from Olkusz. GSTs activity was generally not influenced by zinc. It is possible that GSTs were not the most important target of zinc action. On the contrary, the influence of zinc on CAT activity was found. The increase in CAT activity after zinc treatment was similar for all studied populations. An increase in CAT activity after zinc exposure seems to be the most universal reaction. CAT activity in zinc-treated grasshoppers may explain the mechanism of zinc toxicity based on reactive oxygen forms generation. PMID- 21616165 TI - Offspring production with cryopreserved sperm from a live-bearing fish Xiphophorus maculatus and implications for female fecundity. AB - Xiphophorus fishes are well-established models for biomedical research of spontaneous or induced tumors, and their use in research dates back to the 1930s. Currently, 58 well-pedigreed lines exist among 24 Xiphophorus species housed as live animals at the Xiphophorus Genetic Stock Center. The technique of sperm cryopreservation has been applied to preserve these valuable genetic resources, and production of offspring has been reported with cryopreserved sperm in two species (X. helleri and X. couchianus). The goal of this research was to establish protocols for sperm cryopreservation and artificial insemination that yield live young in X. maculatus, a widely used research species. The objectives were to: 1) collect basic biological characteristics of males, and quantify the sperm production yield after crushing of dissected testis; 2) cryopreserve sperm from X. maculatus by adapting as necessary the protocols for sperm cryopreservation of X. helleri and X. couchianus; 3) use cryopreserved sperm to inseminate virgin females of X maculatus and other species (X. helleri and X. couchianus), and 4) compare experimental trials over a 3-year period to identify opportunities for improving female fecundity. In total, 117 males were used in this study with a standard length of 2.5 +/- 0.3 cm (mean +/- SD), body weight of 0.474 +/- 0.149 g, and dissected testis weight of 7.1 +/- 3.7 mg. Calculation of sperm availability showed 5.9 +/- 2.8 * 10(6) sperm cells per mg of testis weight. Offspring were produced from cryopreserved sperm. Male-to-male variation (1-70%) was observed in post-thaw motility despite little variation in motility before freezing (60-90%) or genetic variation (~100 generations of sib-mating). Comparisons of biological factors of males did not have significant correlations with the production of live young, and the influence of females on production of young was identified from the comparison of artificial insemination over 3 years. Overall, this study describes offspring production from cryopreserved sperm in a third species of Xiphophorus fishes, and identifies the opportunities for improving female fecundity which is essential for establishment of germplasm repositories for Xiphophorus fishes. PMID- 21616167 TI - Evaluation of the direct and indirect response of blood leukocytes to carbon nanotubes (CNTs). AB - Carbon nanotubes (CNTs) possess unique structural and functional properties and are readily internalized by various mammalian cells, making them highly attractive as a tool for gene and drug delivery. However, prior to use in vivo as carriers for therapeutics, their toxicity and potential to elicit an immune response need to be understood. To evaluate the acute response of blood leukocytes to CNTs in vitro, we recreated two specific events: (a) a direct exposure event that may occur due to presence of CNTs in circulation and (b) presentation of CNTs to blood leukocytes via antigen presenting cells. The potential for activation of different leukocyte subpopulations was then evaluated by profiling various early activation markers using flow cytometry. To ensure relevance to gene and drug delivery, these experiments utilized single-walled CNTs (SWCNTs) functionalized with single-stranded (ss)-DNA fragments consisting of guanine-thymine (GT) repeat sequences, which have potential to serve as a backbone for transport of biomolecules and also as a surfactant to prevent aggregation. Results from this study demonstrate that ssDNA-functionalized SWCNTs does not elicit an acute immune response from blood leukocytes through either direct or indirect interactions as verified by the expression of early leukocyte activation markers. FROM THE CLINICAL EDITOR: Carbon nanotubes offer a possible option for targeted gene and drug delivery, but first their toxicity and potential to elicit an immune response need to be understood. The authors of this study demonstrate that ssDNA-functionalized SWCNTs do not elicit an acute immune response from blood leukocytes as verified by the expression of early leukocyte activation markers. PMID- 21616168 TI - Design and potential application of PEGylated gold nanoparticles with size dependent permeation through brain microvasculature. AB - Gold nanoparticles (AuNPs) have gained prominence in several targeting applications involving systemic cancers. Their enhanced permeation and retention within permissive tumor microvasculature provides a selective advantage for targeting. Malignant brain tumors also exhibit transport-permissive microvasculature secondary to blood-brain barrier disruption. Hence AuNPs may have potential relevance for brain tumor targeting. However, there are currently no studies that systematically examine brain microvasculature permeation of polyethylene glycol (PEG)-functionalized AuNPs. Such studies could pave the way for rationale AuNP design for passive targeting of malignant tumors. In this report we designed and characterized AuNPs with varying core particle sizes (4-24 nm) and PEG chain lengths [molecular weight 1000-10,000]. Using an in-vitro model designed to mimic the transport-permissive brain microvasculature, we demonstrate size-dependent permeation properties with respect to core particle size and PEG chain length. In general short PEG chain length (molecular weight 1000-2000) in combination with smallest core size led to optimum permeation in our model system. FROM THE CLINICAL EDITOR: In this report the authors designed and characterized PEGylated gold NPs with varying core particle sizes and PEG chain lengths and demonstrate that short PEG chain length in combination with smallest core size led to optimum permeation of a blood-brain barrier model system. These findings may pave the way to optimized therapy of malignant brain tumors. PMID- 21616169 TI - Bionanotechnologies for treatment and diagnosis of Alzheimer's disease. PMID- 21616170 TI - The effect of dietary flaxseed on improving symptoms of cardiovascular disease in patients with peripheral artery disease: rationale and design of the FLAX-PAD randomized controlled trial. AB - BACKGROUND: Flaxseed is an important source of alpha-linolenic acid an essential omega-3 fatty acid. The possibility that a supplementation of the diet with foods rich in alpha-linolenic acid, antioxidants and fiber (like flaxseed) has not been investigated. METHODS/DESIGN: The primary objective is to determine whether consumption of a diet rich in FLAXseed over a one year period has any beneficial cardiovascular effects in patients with Peripheral Arterial Disease (FLAX-PAD study). This is a single center, prospective, double blinded, randomized controlled clinical trial aimed at in 110 patients over 40 years old and with peripheral arterial disease. Patients will receive 30 g of milled flaxseed (or placebo) per day. Primary endpoints are incidence of myocardial infarction and stroke. Secondary measures include: requirement for surgical interventions, exercise and cardiopulmonary performance, cardiac arrhythmias, serum lipid profile, arterial sufficiency, blood pressure, inflammatory profile, platelet function, changes in drug dosage levels, as well as nutrigenomic and biomarker profiles in the blood. Recruitment and baseline examinations started in October 2008. Baseline data of the 110 patients is shown. CONCLUSIONS: FLAX-PAD will generate data on the safety, tolerability, cardiovascular efficacy and genomic response to a diet rich in flaxseed. It will determine the effects on primary and secondary events (stroke, myocardial infarctions, angina pectoris, cardiac arrhythmias) as well as in secondary endpoints (exercise performance, blood pressure and circulating lipid levels) in patients with PAD. PMID- 21616171 TI - Health, economic, psychological and social impact of educating carers of patients with advanced pulmonary disease (protocol). AB - People with advanced pulmonary disease (APD), such as those with chronic obstructive pulmonary disease, have markedly impaired quality of life. Home Oxygen Therapy (HOT) itself is burdensome, although it often improves survival duration and quality of life in these patients. The exact burdens on informal caregivers of these patients are unknown. The central purpose of the pragmatic randomized controlled study described in this protocol is to determine the effectiveness of improving the skills and knowledge of carers of patients with APD who use HOT. Specifically we aimed to estimate the incremental impact of this carer intervention above usual care on health, economic, psychological and social domains for patient and carer dyads relative to the level of current burden. Eligible patients and their carers were recruited through three major hospitals, and randomized to an intervention or control group. The carers in the intervention group received two home-delivered education sessions based on the principles of academic detailing. Participants are currently being followed over 12 months. The primary outcome will be the proportion of patients surviving without a chronic obstructive pulmonary disease-related readmission / residential (non respite) care over 12 months. Carer secondary outcomes include perceived caregiver burden, level of expected and received social support, perceived level of mastery, self esteem, health related quality of life and disability, and ability to conduct domestic chores and household maintenance, social activities and provide service to others, and fatigue. Secondary patient outcomes include health related quality of life and disability, and current respiratory health status. PMID- 21616172 TI - Are propensity scores really superior to standard multivariable analysis? AB - Clinicians often face difficult decisions despite the lack of evidence from randomized trials. Thus, clinical evidence is often shaped by non-randomized studies exploiting multivariable approaches to limit the extent of confounding. Since their introduction, propensity scores have been used more and more frequently to estimate relevant clinical effects adjusting for established confounders, especially in small datasets. However, debate persists on their real usefulness in comparison to standard multivariable approaches such as logistic regression and Cox proportional hazard analysis. This holds even truer in light of key quantitative developments such as bootstrap and Bayesian methods. This qualitative review aims to provide a concise and practical guide to choose between propensity scores and standard multivariable analysis, emphasizing strengths and weaknesses of both approaches. PMID- 21616173 TI - Oversight and management of a cell therapy clinical trial network: experience and lessons learned. AB - The Cardiovascular Cell Therapy Research Network (CCTRN), sponsored by the National Heart, Lung, and Blood Institute (NHLBI), was established to develop, coordinate, and conduct multiple collaborative protocols testing the effects of cell therapy on cardiovascular diseases. The Network was born into a difficult political and ethical climate created by the recent removal of a dozen drugs from the US formulary and the temporary halting of 27 gene therapy trials due to safety concerns. This article describes the Network's challenges as it initiated three protocols in a polarized cultural atmosphere at a time when oversight bodies were positioning themselves for the tightest vigilance of promising new therapies. Effective strategies involving ongoing education, open communication, and relationship building with the oversight community are discussed. PMID- 21616175 TI - Vaccine candidates for leishmaniasis: a review. AB - Leishmaniasis is a diverse group of clinical syndromes caused by protozoan parasites of the genus Leishmania. The clinical manifestation of the disease varies from self-limiting cutaneous lesions to progressive visceral disease. It is estimated that 350 million people are at risk in 88 countries, with a global incidence of 1-1.5 million cases of cutaneous and 500,000 cases of visceral leishmaniasis. The key control measures mainly rely on early case detection and chemotherapy which has been hampered by the toxicity of drugs, side-effects and by the emergence of drug resistance in parasites. Control of reservoir host and vector is difficult due to operational difficulties and frequent relapses in the host. Therefore, the development of effective and affordable vaccine against leishmaniasis is highly desirable. Although considerable progress has been made over the last decade in understanding immune mechanisms underlying potential candidate antigens, including killed, live attenuated parasites, crude parasites, pure or recombinant Leishmania proteins or DNA encoding leishmanial proteins, as well as immunomodulators from sand fly saliva, very few candidate vaccines have progressed beyond the experimental stage. As such there is no vaccine against any form of human leishmaniasis. In recent years, however, much interest has been stimulated towards vaccination against leishmaniasis focused mainly on cutaneous leishmaniasis with fewer attempts against visceral leishmaniasis. PMID- 21616176 TI - A phytoceramide analog stimulates the production of chemokines through CREB activation in human endothelial cells. AB - Innate immunity is the front-line of self-defense against microbial infection. In mammals, innate immunity interacts with adaptive immunity and has a key role in the regulated immune response. From a pharmaceutical point of view, innate immunity is an ideal target for the development of immunoregulators. Therefore, we aimed to isolate and characterize a novel mammalian immunoregulator isolated from the thermophilic cellulotic fungus Talaromyces sp. 2'-(R)-hydroxy-C(24) phytoceramide (C(24)(2'OH)Phy) was isolated from Talaromyces sp. using a Drosophila ex vivo culture system. C(24)(2'OH)Phy suppressed the immune deficiency (IMD) pathway-dependent expression of antibacterial peptides in Drosophila, whereas it stimulated the production of chemokines in human cells. Structure activity relationship studies of C(24)(2'OH)Phy analogs revealed that both the 2'-(R)-hydroxylignoceroyl group and phytoceramide backbone are essential for the biologic activity of C(24)(2'OH)Phy. Microarray analysis revealed that C(24)(2'OH)Phy selectively activates the transcription of inflammatory response genes, including chemokines. Furthermore, a reporter gene assay and small interfering RNA analysis demonstrated that C(24)(2'OH)Phy stimulates chemokine production through cAMP response element-binding protein activation in human cells. C(24)(2'OH)Phy may be a lead immunostimulating compound in humans. PMID- 21616174 TI - Toll-like receptors are key players in neurodegeneration. AB - The activation of innate immune response is initiated by engagement of pattern recognition receptors (PPRs), such as Toll-like receptors (TLRs). These receptors are expressed in peripheral leukocytes and in many cell types in the central nervous system (CNS). The expression of TLRs in CNS was mainly studied in astrocytes and microglial cells. However, new evidence indicates that these receptors may play an important role in neuronal homeostasis. The expression of TLRs in the CNS is variable and can be modulated by multiple factors, including pro-inflammatory molecules, which are elevated in neurodegenerative diseases and can increase the expression of TLRs in CNS cells. Moreover, activation of TLRs induces the release of pro-inflammatory cytokines. Therefore, TLRs have been shown to play a role in several aspects of neurodegenerative diseases. Here we will discuss results reported in the recent literature concerning the participation of TLRs in neurodegenerative diseases. PMID- 21616177 TI - Dual diaminopimelate biosynthesis pathways in Bacteroides fragilis and Clostridium thermocellum. AB - Bacteroides fragilis and Clostridium thermocellum were recently found to synthesize diaminopimelate (DAP) by way of LL-DAP aminotransferase. Both species also contain an ortholog of meso-diaminopimelate dehydrogenase (Ddh), suggesting that they may have redundant pathways for DAP biosynthesis. The B. fragilis Ddh ortholog shows low homology with other examples of Ddh and this species belongs to a phylum, the Bacteriodetes, not previously known to contain this enzyme. By contrast, the C. thermocellum ortholog is well conserved with known examples of Ddh. Using in vitro and in vivo assays both the B. fragilis and C. thermocellum enzymes were found to be authentic examples of Ddh, displaying kinetic properties typical of this enzyme. The result indicates that B. fragilis contains a sequence diverged form of Ddh. Phylogenomic analysis of the microbial genome database revealed that 77% of species with a Ddh ortholog also contain a second pathway for DAP biosynthesis suggesting that Ddh evolved as an ancillary mechanism for DAP biosynthesis. PMID- 21616178 TI - Mobilization of storage proteins in soybean seed (Glycine max L.) during germination and seedling growth. AB - During germination and early growth of the seedling, storage proteins are degraded by proteases. Currently, limited information is available on the degradation of storage proteins in the soybean during germination. In this study, a combined two-dimensional gel electrophoresis and mass spectrometry approach was utilized to determine the proteome profile of soybean seeds (Glycine max L.; Eunhakong). Comparative analysis showed that the temporal profiles of protein expression are dramatically changed during the seed germination and seedling growth. More than 80% of the proteins identified were subunits of glycinin and beta-conglycinin, two major storage proteins. Most subunits of these proteins were degraded almost completely at a different rate by 120h, and the degradation products were accumulated or degraded further. Interestingly, the acidic subunits of glycinin were rapidly degraded, but no obvious change in the basic chains. Of the five acidic subunits, the degradation of G2 subunit was not apparently affected by at least 96h but the levels decreased rapidly after that, while no newly appearing intermediate was detected upon the degradation of G4 subunit. On the other hand, the degradation of beta-conglycinin during storage protein mobilization appeared to be similar to that of glycinin but at a faster rate. Both alpha and alpha' subunits of beta-conglycinin largely disappeared by 96h, while the beta subunits degraded at the slowest rate. These results suggest that mobilization of subunits of the storage proteins is differentially regulated for seed germination and seedling growth. The present proteomic analysis will facilitate future studies addressing the complex biochemical events taking place during soybean seed germination. PMID- 21616179 TI - R171H missense mutation of INSL6 in a patient with spermatogenic failure. AB - Insulin-like peptide 6 (INSL6) is a newly identified insulin/relaxin family peptide hormone that is predominantly expressed by the male germ cells in testes. A recent murine study demonstrated that INSL6-knockout results in spermatogenic failure. In the present study, human INSL6 gene was screened for mutations that may contribute to human spermatogenic failure. Of 249 patients and 249 healthy control subjects, a heterozygous R171H missense mutation was found in one patient. The R171H mutation probably disturbed the in vivo processing of the INSL6 prohormone because it was located at the absolutely conserved tetrabasic cleavage site between the C-peptide and the A-chain, therefore the R171H missense mutation might be responsible for human spermatogenic failure. PMID- 21616180 TI - Bio-generated metal binding polysaccharides as catalysts for synthetic applications and organic pollutant transformations. AB - Iron and palladium binding an exopolysaccharide (EPS) were obtained and purified from cultures of bacterial cells of Klebsiella oxytoca BAS-10. The strain BAS-10 was able to grow under anaerobic conditions with Fe(III)-citrate as energy and carbon source, producing Fe(III)-EPS that was extracted and used as catalyst in the oxidation reaction of phenol with H(2)O(2). The same bacterial strain was cultivated anaerobically with Na-citrate and Pd(2)(NO)(3) was added during the exponential growth to afford a Pd-EPS, named Bio-Pd (A), that, after isolation and purification, was used as catalyst in the reductive dehalogenation of chlorobenzene as model reaction. For comparison other two palladium binding polysaccharides were prepared: (a) a second type Pd-EPS, named Bio-Pd (B), was obtained by an exchange reaction with Pd acetate starting from an iron-free EPS produced by strain BAS-10 growing on Na-citrate medium; (b) a third type of palladium, named Bio-Pd (C), bound to a different polysaccharide, was recovered after the same exchange reaction applied on glycolipid emulsan obtained from an aerobic culture of Acinetobacter venetianus RAG 1. The superiority of Bio-Pd (A), as catalyst, vs Bio-Pd (B) and (C) was demonstrated. This approach to use microorganisms to prepare metal bound polysaccharides is novel and permits to prepare metal species, sequestrated in aqueous phase that can be useful either as catalysts for synthetic applications or to support the microbial biotransformation of pollutants. PMID- 21616182 TI - Proteomic comparison of virulent phase I and avirulent phase II of Coxiella burnetii, the causative agent of Q fever. AB - Coxiella burnetii, a category B biological warfare agent, causes multiple outbreaks of the zoonotic disease Q fever world-wide, each year. The virulent phase I and avirulent phase II variants of the Nine Mile RSA 493 and 439 strains of C. burnetii were propagated in embryonated hen eggs and then purified by centrifugation through Renografin gradients. Total protein fractions were isolated from each phase and subjected to analysis by one-dimensional electrophoresis plus tandem mass spectrometry. A total of 235 and 215 non redundant proteins were unambiguously identified from the phase I and II cells, respectively. Many of these proteins had not been previously reported in proteomic studies of C. burnetii. The newly identified proteins should provide additional insight into the pathogenesis of Q fever. Several of the identified proteins are involved in the biosynthesis and metabolism of components of the extracellular matrix. Forty-four of the proteins have been annotated as having distinct roles in the pathogenesis or survival of C. burnetii within the harsh phagolysosomal environment. We propose that nine enzymes specifically involved with lipopolysaccharide biosynthesis and metabolism, and that are distinctively present in phase I cells, are virulence-associated proteins. PMID- 21616181 TI - Proteome profiling of wild type and lumican-deficient mouse corneas. AB - To elucidate how the deficiency of a major corneal proteoglycan, lumican, affects corneal homeostasis, we used mass spectrometry to derive the proteome profile of the lumican-deficient and the heterozygous mouse corneas and compared these to the wild type corneal proteome. 2108 proteins were quantified in the mouse cornea. Selected proteins and transcripts were investigated by Western blot and quantitative RT-PCR, respectively. We observed major changes in the composition of the stromal extracellular matrix (ECM) proteins in the lumican-deficient mice. Lumican deficiency altered cellular proteins in the stroma and the corneal epithelium. The ECM changes included increases in fibril forming collagen type I, Collagen type VI, fibromodulin, perlecan, laminin beta2, collagen type IV, nidogen/entactin and anchoring collagen type VII in the Lum+/- and the Lum-/- mouse corneas, while the stromal proteoglycans decorin, biglycan and keratocan were decreased in the Lum-/-( corneas. Cellular protein changes included increases in alcohol dehydrogenase, superoxide dismutase and decreases in epithelial cytokeratins 8 and 14. We also detected proteins that are novel to the cornea. The proteomes will provide an insight into the lumican-deficient corneal phenotype of stromal thinning and loss of transparency and a better understanding of pathogenic changes in corneal and ocular dystrophies. PMID- 21616183 TI - Deconvolution of overlapping isotopic clusters improves quantification of stable isotope-labeled peptides. AB - High-resolution mass spectrometry and the use of stable isotopes have greatly improved our ability to quantify proteomes. Typically, the relative abundance of peptides is estimated by identifying the isotopic clusters and by comparing the peak intensities of peptide pairs. However, when the mass shift between the labeled peptides is small, there can be the possibility for overlap of the isotopic clusters which will hamper quantification accuracy with a typical upwards bias for the heavier peptide. Here, we investigated the impact of the overlapping peak issue with respect to dimethyl based quantification and we confirmed there can be need for correction. In addition, we present a tool that can correct overlapping issues when they arise which is based on modeling isotopic distributions. We demonstrate that our approach leads to improved accuracy and precision of protein quantification. PMID- 21616184 TI - Functions of Kaposi's sarcoma-associated herpesvirus microRNAs. AB - MicroRNAs can modulate gene expression post-transcriptionally by altering mRNA stability and protein translation. Multiple DNA viruses express viral microRNAs and have been shown to modulate expression of host and viral genes. Through various methods of microRNA target identification, we are beginning to understand the various roles of viral miRNAs including viral replication, immune evasion and apoptosis. This review is focused on the roles of microRNAs expressed by Kaposi's sarcoma-associated herpesvirus or human herpesvirus 8. This article is part of a Special Issue entitled: "MicroRNAs in viral gene regulation". PMID- 21616185 TI - Diversity of operation in ATP-dependent chromatin remodelers. AB - Chromatin is actively restructured by a group of proteins that belong to the family of ATP-dependent DNA translocases. These chromatin remodelers can assemble, relocate or remove nucleosomes, the fundamental building blocks of chromatin. The family of ATP-dependent chromatin remodelers has many properties in common, but there are also important differences that may account for their varying roles in the cell. Some of the important characteristics of these complexes have begun to be revealed such as their interactions with chromatin and their mechanism of operation. The different domains of chromatin remodelers are discussed in terms of their targets and functional roles in mobilizing nucleosomes. The techniques that have driven these findings are discussed and how these have helped develop the current models for how nucleosomes are remodeled. This article is part of a Special Issue entitled: Snf2/Swi2 ATPase structure and function. PMID- 21616188 TI - Achieving nutrient density: a vegetarian approach. PMID- 21616187 TI - MicroRNA-like antivirals. AB - Employing engineered DNA templates to express antiviral microRNA (miRNA) sequences has considerable therapeutic potential. The durable silencing that may be achieved with these RNAi activators is valuable to counter chronic viral infections, such as those caused by HIV-1, hepatitis B, hepatitis C and dengue viruses. Early use of expressed antiviral miRNAs entailed generation of cassettes containing Pol III promoters (e.g. U6 and H1) that transcribe virus-targeting short hairpin RNA mimics of precursor miRNAs. Virus escape from single gene silencing elements prompted later development of combinatorial antiviral miRNA expression cassettes that form multitargeting siRNAs from transcribed long hairpin RNA and polycistronic primary miRNA sequences. Weaker Pol III and Pol II promoters have also been employed to control production of antiviral miRNA mimics, improve dose regulation and address concerns about toxicity caused by saturation of the endogenous miRNA pathway. Efficient delivery of expressed antiviral sequences remains challenging and utilizing viral vectors, which include recombinant adenoviruses, adeno-associated viruses and lentiviruses, has been favored. Investigations using recombinant lentiviruses to transduce CD34+ hematological precursor cells with expressed HIV-1 gene silencers are at advanced stages and show promise in preclinical and clinical trials. Although the use of expressed antiviral miRNA sequences to treat viral infections is encouraging, eventual therapeutic application will be dependent on rigorously proving their safety, efficient delivery to target tissues and uncomplicated large scale preparation of vector formulations. This article is part of a special issue entitled: MicroRNAs in viral gene regulation. PMID- 21616189 TI - Introducing ADA's 86th President: Sylvia Escott-Stump, MA, RD, FADA, LDN. PMID- 21616190 TI - Credibility: becoming a trusted health professional. PMID- 21616186 TI - Regulation of cellular miRNA expression by human papillomaviruses. AB - High-risk HPV infection leads to aberrant expression of cellular oncogenic and tumor suppressive miRNAs. A large number of these miRNA genes are downstream targets of the transcription factors c-Myc, p53, and E2F and their expression can therefore be modulated by oncogenic HPV E6 and E7. Cervical cancer represents a unique tumor model for understanding how viral E6 and E7 oncoproteins deregulate the expression of the miR-15/16 cluster, miR-17-92 family, miR-21, miR-23b, miR 34a, and miR-106b/93/25 cluster via the E6-p53 and E7-pRb pathways. Moreover, miRNAs may influence the expression of papillomavirus genes in a differentiation dependent manner by targeting viral RNA transcripts. Cellular miRNAs affecting HPV DNA replication are of great interest and will be a future focus. We are entering an era focusing on miRNA and noncoding RNA, and the studies on HPV and host miRNA interactions will continue shedding more light on our understanding of the HPV life cycle and the mechanistic underpinnings of HPV-induced oncogenesis. This article is part of a Special Issue entitled: "MicroRNAs in viral gene regulation". PMID- 21616191 TI - Functional foods: consumer attitudes, perceptions, and behaviors in a growing market. PMID- 21616192 TI - Expanding opportunities for registered dietitians in the health care system. PMID- 21616193 TI - A vegetarian diet for weight management. PMID- 21616194 TI - A vegetarian dietary pattern as a nutrient-dense approach to weight management: an analysis of the national health and nutrition examination survey 1999-2004. AB - BACKGROUND: Population-based studies have shown that vegetarians have lower body mass index than nonvegetarians, suggesting that vegetarian diet plans may be an approach for weight management. However, a perception exists that vegetarian diets are deficient in certain nutrients. OBJECTIVE: To compare dietary quality of vegetarians, nonvegetarians, and dieters, and to test the hypothesis that a vegetarian diet would not compromise nutrient intake when used to manage body weight. DESIGN: Cross-sectional analysis of National Health and Nutrition Examination Survey (1999-2004) dietary and anthropometric data. Diet quality was determined using United States Department of Agriculture's Healthy Eating Index 2005. Participants included adults aged 19 years and older, excluding pregnant and lactating women (N = 13,292). Lacto-ovo vegetarian diets were portrayed by intakes of participants who did not eat meat, poultry, or fish on the day of the survey (n = 851). Weight-loss diets were portrayed by intakes of participants who consumed 500 kcal less than their estimated energy requirements (n = 4,635). Mean nutrient intakes and body mass indexes were adjusted for energy, sex, and ethnicity. Using analysis of variance, all vegetarians were compared to all nonvegetarians, dieting vegetarians to dieting nonvegetarians, and nondieting vegetarians to nondieting nonvegetarians. RESULTS: Mean intakes of fiber, vitamins A, C, and E, thiamin, riboflavin, folate, calcium, magnesium, and iron were higher for all vegetarians than for all nonvegetarians. Although vegetarian intakes of vitamin E, vitamin A, and magnesium exceeded that of nonvegetarians (8.3 +/- 0.3 vs 7.0 +/- 0.1 mg; 718 +/- 28 vs 603 +/- 10 MUg; 322 +/- 5 vs 281 +/ 2 mg), both groups had intakes that were less than desired. The Healthy Eating Index score did not differ for all vegetarians compared to all nonvegetarians (50.5 +/- 0.88 vs 50.1 +/- 0.33, P = 0.6). CONCLUSIONS: These findings suggest that vegetarian diets are nutrient dense, consistent with dietary guidelines, and could be recommended for weight management without compromising diet quality. PMID- 21616196 TI - Decreased salt intake in Japanese men aged 40 to 70 years and women aged 70 to 79 years: an 8-year longitudinal study. AB - BACKGROUND: It is not known whether salt intake decreases over time in the same population. This study attempts to describe salt intake for 8 years according to age groups, and examines whether salt intake changes over time in community dwelling middle-aged and elderly Japanese subjects. METHODS: Data were collected as part of the National Institute for Longevity Sciences Longitudinal Study of Aging. Participants included 544 men and 512 women who participated in and completed all nutrition surveys from the first (1997-2000) to fifth (2006-2008) study waves. Each study wave was conducted for 2 years; in individuals, the entire follow-up period was 8 years. Salt and energy intake were calculated from 3-day diet records with photographs. The mixed-effects regression model was used for analysis of repeated measures of salt intake. RESULTS: Mean age and salt intake for study participants at first participation in the survey were 56.5 +/- 9.3 years and 12.8 +/- 3.3 g/day in men and 55.8 +/- 9.4 years and 10.6 +/- 2.5 g/day in women, respectively. Mean energy intake decreased in men and women in all age groups from the first to fifth study waves. Eight-year longitudinal data showed that salt intake decreased in men. In stratified analyses by age, mean salt intake in men decreased 0.08 g/year among 40- to 49-year-olds, 0.09 g/year among 50- to 59-year-olds, 0.16 g/year among 60- to 69-year-olds, and 0.14 g/year among 70- to 79-year-olds. For women, mean salt intake decreased 0.08 g/year among 70- to 79-year-olds (P = 0.098). After adjusting for energy intake, salt intake was decreased among 60- to 69-year-old men (P = 0.049) and increased among 50- to 59-year-old women (P = 0.015). CONCLUSIONS: Absolute salt intake was decreased among all age groups from 40 to 70 years in men and from 70 to 79 years in women. An increased focus on reducing energy intake resulted in only a modest decrease in salt intake. Although we observed a decline, salt intake still exceeded recommended levels. Efforts that focus on salt reduction are needed to address this important public health problem. PMID- 21616195 TI - A structured diet and exercise program promotes favorable changes in weight loss, body composition, and weight maintenance. AB - BACKGROUND: A number of diet and exercise programs purport to help promote and maintain weight loss. However, few studies have compared the efficacy of different methods. OBJECTIVE: To determine whether adherence to a meal replacement-based diet program (MRP) with encouragement to increase physical activity is as effective as following a more structured meal-plan-based diet and supervised exercise program (SDE) in sedentary obese women. DESIGN: Randomized comparative effectiveness trial. PARTICIPANTS/SETTING: From July 2007 to October 2008, 90 obese and apparently healthy women completed a 10-week university-based weight loss trial while 77 women from this cohort also completed a 24-week weight maintenance phase. INTERVENTION: Participants were matched and randomized to participate in an MRP or SDE program. MAIN OUTCOME MEASURES: Weight loss, health, and fitness-related data were assessed at 0 and 10 weeks on all subjects as well as at 14, 22, and 34 weeks on participants who completed the weight maintenance phase. STATISTICAL ANALYSES PERFORMED: Data were analyzed by multivariate analysis of variance for repeated measures. RESULTS: During the 10-week weight loss phase, moderate and vigorous physical activity levels were significantly higher in the SDE group with no differences observed between groups in daily energy intake. The SDE group lost more weight (-3.1 +/- 3.7 vs -1.6 +/- 2.5 kg; P = 0.03); fat mass (-2.3 +/- 3.5 vs -0.9 +/- 1.6 kg; P = 0.02); centimeters from the hips (-4.6 +/- 7 vs -0.2 +/- 6 cm; P = 0.002) and waist (-2.9 +/- 6 vs -0.6 +/- 5 cm; P = 0.05); and, experienced a greater increase in peak aerobic capacity than participants in the MRP group. During the 24-week maintenance phase, participants in the SDE group maintained greater moderate and vigorous physical activity levels, weight loss, fat loss, and saw greater improvement in maximal aerobic capacity and strength. CONCLUSIONS: In sedentary and obese women, an SDE based program appears to be more efficacious in promoting and maintaining weight loss and improvements in markers of health and fitness compared to an MRP type program with encouragement to increase physical activity. PMID- 21616197 TI - Competitive foods sales are associated with a negative effect on school finances. AB - BACKGROUND: It is widely presumed that competitive foods-foods offered for sale in schools in addition to reimbursable federal meals programs-provide revenue that is essential to maintain school foodservices. However, evidence is lacking to demonstrate whether competitive foods sales truly improve foodservice financial viability. OBJECTIVE: The aim of this research was to assess whether or not competitive foods sales have an overall positive financial effect on school foodservice finances. DESIGN AND STATISTICAL ANALYSES: This observational study used a multivariate time series analysis of annual foodservice financial data from repeated observations of 344 Minnesota public school districts between 2001 and 2008 (N=2,695). First, revenue from competitive foods was assessed in terms of whether or not such revenue displaced or complemented revenue from reimbursable meals. Second, profit from competitive foods was assessed in terms of whether or not such profit displaced or increased total school foodservice profit. RESULTS: Fixed effects models indicated small but significant negative relationships between competitive foods sales and reimbursable meals revenue, as well as overall foodservice profit. A 10% increase in competitive foods revenue was associated with a 0.1% decrease in reimbursable meals revenue (P<0.05). A 10% increase in competitive foods profit was associated with a 0.7% decrease in overall foodservice profit among schools with profitable competitive sales (P<0.10). CONCLUSIONS: Study findings suggest that competitive foods can have a negative effect on school foodservice finances. Better understanding of foodservice finances could influence current approaches to improve school nutrition. Improved recordkeeping may be necessary to ensure that public funds are not used to subsidize schools' competitive offerings. PMID- 21616198 TI - Travel history, hunting, and venison consumption related to prion disease exposure, 2006-2007 FoodNet Population Survey. AB - The transmission of bovine spongiform encephalopathy (BSE) to human beings and the spread of chronic wasting disease (CWD) among cervids have prompted concerns about zoonotic transmission of prion diseases. Travel to the United Kingdom and other European countries, hunting for deer or elk, and venison consumption could result in the exposure of US residents to the agents that cause BSE and CWD. The Foodborne Diseases Active Surveillance Network 2006-2007 population survey was used to assess the prevalence of these behaviors among residents of 10 catchment areas across the United States. Of 17,372 survey respondents, 19.4% reported travel to the United Kingdom since 1980, and 29.5% reported travel to any of the nine European countries considered to be BSE-endemic since 1980. The proportion of respondents who had ever hunted deer or elk was 18.5%, and 1.2% had hunted deer or elk in a CWD-endemic area. More than two thirds (67.4%) reported having ever eaten deer or elk meat. Respondents who traveled spent more time in the United Kingdom (median 14 days) than in any other BSE-endemic country. Of the 11,635 respondents who had consumed venison, 59.8% ate venison at most one to two times during their year of highest consumption, and 88.6% had obtained all of their meat from the wild. The survey results were useful in determining the prevalence and frequency of behaviors that could be important factors for foodborne prion transmission. PMID- 21616199 TI - Signaling proteins that influence energy intake may affect unintentional weight loss in elderly persons. AB - After age 70 to 75 years, average body weight decreases both in ailing and healthy people because of a loss of appetite that results in reduced energy intake and the loss of body fat and lean muscle tissue. This so-called anorexia of aging predisposes elderly people to continued pathologic weight loss and malnutrition-major causes of morbidity and mortality. Health care professionals must understand the many factors involved in the anorexia of aging to help older adults prevent unintentional weight loss. Psychological, social, and cultural factors are important effectors; however, physiological factors are emphasized here because they are not thoroughly understood and they make it inherently difficult for most people to alter their body weight. Monoamines, steroid hormones (glucocorticoids and mineralocorticoids), endocannabinoids, and proteins all influence body weight. This review is an analysis of proteins from the brain, pancreas, adipose tissue, and gastrointestinal tract that are known to affect energy intake and energy balance, with an attempt to identify those factors that may change with aging. The articles included in this review were obtained by a PubMed database search using the keywords mouse OR rat OR human AND aged OR aging OR older OR elderly AND adult AND anorexia OR "unintentional weight loss," and each of the individual proteins discussed, as well as from the reference lists of those articles. The results reveal that some proteins may be important in the development of unintentional weight loss in elderly persons, whereas others may not have a significant role. However, many of the proteins that could conceivably have a role in unintentional weight loss have not yet been studied with that question in mind. Preventing unintentional weight loss in older adults is an important goal and further research on the role of proteins important for the maintenance of energy balance and the development of unintentional weight loss in elderly persons is warranted. PMID- 21616200 TI - Association of delta13C in fingerstick blood with added-sugar and sugar-sweetened beverage intake. AB - A reliance on self-reported dietary intake measures is a common research limitation, thus the need for dietary biomarkers. Added-sugar intake may play a role in the development and progression of obesity and related comorbidities; common sweeteners include corn and sugar cane derivatives. These plants contain a high amount of 13C, a naturally occurring stable carbon isotope. Consumption of these sweeteners, of which sugar-sweetened beverages are the primary dietary source, might be reflected in the delta13C value of blood. Fingerstick blood represents an ideal substrate for bioassay because of its ease of acquisition. The objective of this investigation was to determine if the delta13C value of fingerstick blood is a potential biomarker of added-sugar and sugar-sweetened beverage intake. Individuals aged 21 years and older (n = 60) were recruited to attend three laboratory visits; assessments completed at each visit depended upon a randomly assigned sequence (sequence one or two). The initial visit included assessment of height, weight, and dietary intake (sequence one: beverage intake questionnaire, sequence two: 4-day food intake record). Sequence one participants completed a food intake record at visit two, and nonfasting blood samples were obtained via routine fingersticks at visits one and three. Sequence two participants completed a beverage intake questionnaire at visit two, and provided fingerstick blood samples at visits two and three. Samples were analyzed for delta13C value using natural abundance stable isotope mass spectrometry. delta13C value was compared to dietary outcomes in all participants, as well as among those in the highest and lowest tertile of added-sugar intake. Reported mean added-sugar consumption was 66 +/- 5 g/day, and sugar-sweetened beverage consumption was 330 +/- 53 g/day and 134 +/- 25 kcal/day. Mean fingerstick delta13C value was -19.940/00 +/- 0.100/00, which differed by body mass index status. delta13C value was associated (all P < 0.05) with intake of total added sugars (g, r = 0.37; kcal, r = 0.37), soft drinks (g, r = 0.26; kcal, r = 0.27), and total sugar-sweetened beverage (g, r = 0.28; kcal, r = 0.35). The delta13C value in the lowest and the highest added-sugar intake tertiles were significantly different (mean difference = -0.480/00; P = 0.028). Although there are several potential dietary sources for blood carbon, the delta13C value of fingerstick blood shows promise as a noninvasive biomarker of added-sugar and sugar-sweetened beverage intake based on these findings. PMID- 21616202 TI - The children's menu assessment: development, evaluation, and relevance of a tool for evaluating children's menus. AB - Restaurant foods represent a substantial portion of children's dietary intake, and consumption of foods away from home has been shown to contribute to excess adiposity. This descriptive study aimed to pilot-test and establish the reliability of a standardized and comprehensive assessment tool, the Children's Menu Assessment, for evaluating the restaurant food environment for children. The tool is an expansion of the Nutrition Environment Measures Survey-Restaurant. In 2009-2010, a randomly selected sample of 130 local and chain restaurants were chosen from within 20 miles of Little Rock, AR, to examine the availability of children's menus and to conduct initial calibration of the Children's Menu Assessment tool (final sample: n=46). Independent raters completed the Children's Menu Assessment in order to determine inter-rater reliability. Test-retest reliability was also examined. Inter-rater reliability was high: percent agreement was 97% and Spearman correlation was 0.90. Test-retest was also high: percent agreement was 91% and Spearman correlation was 0.96. Mean Children's Menu Assessment completion time was 14 minutes, 56 seconds +/- 10 minutes, 21 seconds. Analysis of Children's Menu Assessment findings revealed that few healthier options were available on children's menus, and most menus did not provide parents with information for making healthy choices, including nutrition information or identification of healthier options. The Children's Menu Assessment tool allows for comprehensive, rapid measurement of the restaurant food environment for children with high inter-rater reliability. This tool has the potential to contribute to public health efforts to develop and evaluate targeted environmental interventions and/or policy changes regarding restaurant foods. PMID- 21616201 TI - Demographic and financial characteristics of school districts with low and high a la Carte sales in rural Kansas Public Schools. AB - Reducing a la carte items in schools-foods and beverages sold outside the reimbursable meals program-can have important implications for childhood obesity. However, schools are reluctant to reduce a la carte offerings because of the impact these changes could have on revenue. Some foodservice programs operate with limited a la carte sales, but little is known about these programs. This secondary data analysis compared rural and urban/suburban school districts with low and high a la carte sales. Foodservice financial records (2007-2008) were obtained from the Kansas State Department of Education for all public K-12 school districts (n=302). chi2 and t tests were used to examine the independent association of variables to a la carte sales. A multivariate model was then constructed of the factors most strongly associated with low a la carte sales. In rural districts with low a la carte sales, lunch prices and participation were higher, lunch costs and a la carte quality were lower, and fewer free/reduced price lunches were served compared to rural districts with high a la carte sales. Lunch price (odds ratio=1.2; 95% confidence interval, 1.1 to 1.4) and free/reduced price lunch participation (odds ratio=3.0; 95% confidence interval, 1.0 to 9.8) remained in the multivariate model predicting low a la carte sales. No differences were found between urban/suburban districts with low and high a la carte sales. Findings highlight important factors to maintaining low a la carte sales. Schools should consider raising lunch prices and increasing meal participation rates as two potential strategies for reducing the sale of a la carte items without compromising foodservice revenue. PMID- 21616203 TI - Perceived environmental church support is associated with dietary practices among African-American adults. AB - A unique strength of the African-American community is the importance of church and faith. Interventions promoting health might want to build on these strengths by developing faith-based interventions that encourage churches to create an environment that supports behavior change. The objective of the study was to examine the relationship between perceived environmental church support for healthy eating and intake of fruit and vegetables and fat- and fiber-related behaviors, and to examine whether these relationships differ by sex. The design was a cross-sectional study in which participants completed self-report dietary and perceived church support measures before initiation of an intervention. Relationships between fruit and vegetable consumption, fat- and fiber-related behaviors, and perceived church support (eg, total, written informational, spoken informational, instrumental [fruit and vegetable consumption only]), along with Support*Sex interactions were examined. Participants were 1,136 African-American church members from four geographically defined districts in South Carolina. Statistical analyses included regression models controlling for sex, age, years of education, health rating, and body mass index using SAS PROC MIXED. A separate model was conducted for each measure of perceived church support and each type of healthy eating index. Perceived total church support and perceived written and spoken informational church support were associated with considerably higher fruit and vegetable intake and more favorable fiber-related behaviors, whereas only perceived total and perceived written informational support were associated with more low-fat dietary behaviors. Perceived instrumental church support was not associated with fruit and vegetable consumption. No sex differences were found. The social and physical church environment can be an important factor influencing the dietary habits of its members. Future faith-based interventions should further explore the role of the church environment in improving the dietary practices of its members. PMID- 21616204 TI - Establishing a baseline measure of school wellness-related policies implemented in a nationally representative sample of school districts. AB - The Child Nutrition and WIC Reauthorization Act of 2004 required school districts to establish a local school wellness policy by the first day of the 2006-2007 school year. To provide a baseline measure of the extent to which wellness related policies were implemented in school districts nationwide in 2006, this study analyzed data from the 2006 School Health Policies and Programs Study (SHPPS). SHPPS used a cross-sectional design to measure policies and practices among a nationally representative sample of 538 public school districts. The authors applied a standardized wellness policy coding system to the data by matching each element to relevant questions from SHPPS and calculated the percentage of school districts meeting each element in the coding system. Statistical analyses included calculation of 95% confidence intervals for percentages and mean number of elements met in each area. In 2006, none of the districts met all elements included in the coding system for local wellness policies. In addition, the percentage of districts meeting each element varied widely. On average, districts met the greatest number of elements in the area of nutrition education and the least number of elements in the area of physical activity. By applying a coding system for district policies to an existing dataset, this study used a novel approach to determine areas of strength and weakness in the implementation of local school wellness-related policies in 2006. PMID- 21616205 TI - American Dietetic Association: standards of practice and standards of professional performance for registered dietitians (competent, proficient, and expert) in integrative and functional medicine. PMID- 21616206 TI - Entry-level dietetics practice today: results from the 2010 commission on dietetic registration entry-level dietetics practice audit. PMID- 21616207 TI - Is the incidence of foodborne illness in the United States changing? PMID- 21616208 TI - Safety of vitrectomy for floaters--how safe is safe? PMID- 21616209 TI - Preoperative evaluation for patients with choroidal/ciliary body melanomas: what is necessary? PMID- 21616210 TI - Access to cataract surgical services: international ophthalmology accepts the challenge. PMID- 21616212 TI - Dome-shaped macula. PMID- 21616213 TI - Topography-guided conductive keratoplasty for keratoconus. PMID- 21616215 TI - Prognosis of upper eyelid epiblepharon repair in Down syndrome. PMID- 21616217 TI - Prognostic factors and outcomes in endogenous Klebsiella endophthalmitis. PMID- 21616219 TI - Endothelial cell density after Descemet membrane endothelial keratoplasty: 1- to 4-year follow-up. PMID- 21616220 TI - Bronchomegaly as a complication of fetal endoscopic tracheal occlusion. A caution and a possible solution. AB - Fetal medicine is developing rapidly and aims to improve the outcome for fetuses with congenital anomalies. Fetal endoscopic tracheal occlusion (FETO) has been developed for fetuses with congenital diaphragmatic hernia to counterbalance the compression of the lung by the abdominal viscera, preserving the pulmonary maturation. Because the perinatal morbidity and mortality of patients treated with FETO have decreased, new complications are emerging in the older survivors. Tracheomegaly has been reported to be a late complication of FETO, sometimes requiring tracheostomy. We report a case of bronchial dilatation after FETO and suggest an alternative surgical treatment. PMID- 21616221 TI - Difficult central venous access removal: case reports of the use of endovascular snare shearing of endothelialized tetherings. AB - Although a fibrin sheath occurs in most long-standing central venous catheters, they do not typically interfere with complete removal of the catheter. We present 2 cases of long-standing catheters that could not be removed with simple surgical techniques because of endotheliazation via fibrous attachments to the venous wall. Both catheters were successfully removed using a modified snare technique through the right femoral vein. PMID- 21616222 TI - Dieulafoy lesion in the ileum of a child: a case report. AB - A 14-year-old girl had massive bleeding from a Dieulafoy lesion of the ileum. A preoperative dynamic computed tomography scan detected the point of bleeding. Selective arteriography with embolization using microcoils could not stop the bleeding, but the microcoils were useful as markers of the location of the bleeding point. The position of the microcoils was confirmed by intraoperative fluoroscopy. A partial resection of the ileum that included the lesion was performed. The pathologic finding was Dieulafoy lesion of the ileum. Dieulafoy lesion is a rare condition that usually presents in the stomach. Dieulafoy lesion of the ileum is extraordinarily rare, and to our knowledge, this is only the second case report of an ileal lesion in a child. PMID- 21616223 TI - Lipoblastoma: a rare lesion in the differential diagnosis of childhood mediastinal tumors. AB - Lipoblastoma is a rare, benign, fatty tissue tumor that occurs in childhood. The location of this tumor in the mediastinum and extension to the chest wall is uncommon. We describe a 12-month-old male infant with a mediastinal lipoblastoma discovered because of a chest wall swelling. Computed tomography showed the deep component and fatty content of the lesion suggestive of the diagnosis. Total excision of the mass was carried out. The histologic examination of the lesion confirmed the diagnosis of lipoblastoma. Mediastinal lipoblastoma should be considered among the possible diagnoses of a fatty mediastinal tumor in childhood. PMID- 21616224 TI - Double duodenal atresia with perforation: a case report. AB - Duodenal atresia is a common cause of neonatal bowel obstruction. Double duodenal atresia has been reported as a rare variation of duodenal atresia. This report discusses the importance of an appreciation of unusual anatomy in neonatal duodenal obstruction and highlights the importance of assessing bowel continuity intraoperatively. PMID- 21616225 TI - Intrapericardial diaphragmatic hernia: a rare type of congenital diaphragmatic hernia. AB - An extremely rare case of congenital intrapericardial diaphragmatic hernia is presented. Since 1981, only 14 cases have been reported in the literature. A 5 year-old girl presented with dyspnea on exertion and easy fatigability. Computed tomography was suggestive of an anterior diaphragmatic hernia. Laparoscopy followed by successful open repair of hernia was performed. PMID- 21616226 TI - Enlarging unilateral breast mass in an adolescent male: an unusual presentation of intraductal papilloma. AB - Breast masses presenting in adolescent boys are rare and are almost uniformly owing to gynecomastia. Although surgical referral for breast masses in adolescent boys is common, intervention is typically for cosmesis. We report the case of a 14-year-old boy who presented with an enlarging unilateral breast mass, which was found to be owing to an intraductal papilloma at the time of surgical excision. PMID- 21616227 TI - Enhancing safety of laparoscopic vascular control for neonatal sacrococcygeal teratoma. AB - Life-threatening bleeding is a hazard of major tumor excision in children. However, fatalities from inadvertent arterial ligation should not be overlooked. Sacrococcygeal teratoma is the commonest neonatal tumor. Laparotomy to ligate the median sacral artery has been used to preempt potentially fatal resectional bleeding. Use of laparoscopy to achieve the same is an evolving technique, with only 7 neonatal cases described. As such, the Idea, Development, Exploration, Assessment, Long-term study (IDEAL) guidelines on surgical innovation recommend case reports addressing proof of concept, technical factors and safety tips. Fortunately, mistaken arterial division is so far unreported during laparoscopic median sacral artery ligation. However, as uptake widens, anatomical distortion by tumor and surgeon disorientation at endosurgery are risk factors for even such inconceivable complications. We report a successful case of laparoscopic vascular control for neonatal sacrococcygeal teratoma and demonstrate an observation that serves as a useful safety check for this procedure (as well as the open alternative). PMID- 21616228 TI - Laparoscopic repair of traumatic abdominal wall hernia from handlebar injury. AB - A 14-year-old boy was seen at an outside hospital after falling over the handlebar of his bicycle and was discharged home. He was subsequently seen in our emergency department with complaints of persistent abdominal pain. A computed tomography scan of the abdomen revealed disruption of the muscles of the upper right abdominal wall containing the hepatic flexure of the colon, with a small amount of intraperitoneal free fluid noted. The patient underwent laparoscopic exploration using 3 ports (2-5 mm and 1-12 mm) and 2 separate stab incisions. The traumatic abdominal wall hernia was repaired with interrupted sutures placed with an ENDO CLOSE (Covidien, Mansfield, MA) device, and a mesenteric defect in the colon was approximated with intracorporeal sutures. The trocar sites were sutured closed. The patient recovered well and was discharged home. Follow-up examination revealed no abdominal wall defect and resolution of his symptoms. Laparoscopic repair of a traumatic abdominal wall defect and exploratory laparoscopy after trauma is feasible and safe in the pediatric patient. It should be considered as an alternative approach with potentially less morbidity than an exploratory laparotomy for handlebar injuries in a stable patient. PMID- 21616229 TI - Noblesse oblige - the pediatric surgeon (h)as the key to quality control and improvement. The Fred MacLoed Lecture. AB - The trust crisis in health care demands action by health care professionals. Trust is based on quality, candor, and accountability. The pediatric surgeon, as the expert in the field, should be in control of quality management. By improving quality, the trust in the health care system can be restored. Quality is defined as being on target with minimal variation. To assess these targets, performance indicators have been developed by the Association of Pediatric Surgeons in The Netherlands for 7 neonatal conditions. Variation can be distinguished as special cause and common-cause variation using the control chart method of Walter Shewhart. The various activities in this field that have been developed and are ongoing in The Netherlands are presented. PMID- 21616230 TI - Institutional practice and outcome variation in the management of congenital diaphragmatic hernia and gastroschisis in Canada: a report from the Canadian Pediatric Surgery Network. AB - BACKGROUND: Perinatal management of congenital diaphragmatic hernia (CDH) and gastroschisis (GS) remains nonstandardized and institution specific. This analysis describes practice and outcome variation across a national network. METHODS: A national, prospective, disease-specific database for CDH and GS was evaluated over 4 years. Centers were evaluated individually and defined as low (low-volume center [LVC]) or high (high-volume center [HVC]) volume based on case mean. RESULTS: Congenital diaphragmatic hernia. Two hundred fifteen liveborn cases were studied (mean, 14.3 cases/center) across 15 centers (8 LVCs and 7 HVCs). Significant interinstitutional practice variation was noted in rates of termination (0%-40%) and cesarean delivery (0%-61%). Centers demonstrated marked variation in ventilation strategies, vasodilator and paralytic use, timing of surgery, and rates of primary closure. Overall survival was 81.4% (LVC, 76.9%; HVC, 82.4%; P = .43). Gastroschisis. Four hundred sixteen cases were investigated (mean, 26 cases/center; range, 6-72) across 16 centers (10 LVCs and 6 HVCs). Cesarean delivery rates varied widely between centers (0%-86%) as did timing of closure (early vs delayed, 1%-100%). There was no difference in length of stay, days on total parenteral nutrition, and overall survival (94.3% vs 97.2%; P = .17) between LVCs and HVCs. CONCLUSIONS: The existence of perinatal practice and outcome variation for GS and CDH suggests targets for improved delivery of care and justifies efforts to standardize treatment on a national basis. PMID- 21616231 TI - Impact of prenatal evaluation and protocol-based perinatal management on congenital diaphragmatic hernia outcomes. AB - BACKGROUND/PURPOSE: Although intuitive, the benefit of prenatal evaluation and multidisciplinary perinatal management for fetuses with congenital diaphragmatic hernia (CDH) is unproven. We compared the outcome of prenatally diagnosed patients with CDH whose perinatal management was by a predefined protocol with those who were diagnosed postnatally and managed by the same team. We hypothesized that patients with CDH undergoing prenatal evaluation with perinatal planning would demonstrate improved outcome. METHODS: Retrospective chart review of all patients with Bochdalek-type CDH at a single institution between 2004 and 2009 was performed. Patients were stratified by history of perinatal management, and data were analyzed by Fisher's Exact test and Student's t test. RESULTS: Of 116 patients, 71 fetuses presented in the prenatal period and delivered at our facility (PRE), whereas 45 infants were either outborn or postnatally diagnosed (POST). There were more high-risk patients in the PRE group compared with the POST group as indicated by higher rates of liver herniation (63% vs 36%, P = .03), need for patch repair (57% vs 27%, P = .004), and extracorporeal membrane oxygenation use (35% vs 18%, P = .05). Despite differences in risk, there was no difference in 6-month survival between groups (73% vs 73%). CONCLUSIONS: Patients with CDH diagnosed prenatally are a higher risk group. Prenatal evaluation and multidisciplinary perinatal management allows for improved outcome in these patients. PMID- 21616232 TI - Influence of location of delivery on outcome in neonates with congenital diaphragmatic hernia. AB - BACKGROUND: Although it is often recommended that infants with antenatally diagnosed congenital diaphragmatic hernia (CDH) be delivered in a perinatal center, this practice has not been scientifically validated. METHODS: Data were obtained from The Canadian Pediatric Surgery Network, covering 16 pediatric surgical centers over a 4-year period. Inborn was defined as birth in a hospital with a neonatal intensive care unit or connected to a neonatal intensive care unit by a bridge or tunnel. Outborn was defined as requiring transfer by ambulance or flight. Primary outcome variable was mortality. RESULTS: Of 140 infants with antenatally diagnosed CDH, 75 were inborn and 65 were outborn. Univariate analysis demonstrated no significant difference between groups with respect to gestational age, birth weight, days to surgery, primary repair, need for ventilation, use of pressors or extracorporeal membrane oxygenation, or incidence of comorbidities. Severity of illness, as reflected by the Score for Neonatal Acute Physiology II (SNAP II), was significantly higher among inborn infants (21 [interquartile range, 7-32] vs 5 [interquartile range, 9-12]; P = .0001). Logistic regression analysis, controlling for severity of illness, revealed that location of delivery was a significant independent predictor for mortality, with an odds ratio of dying when outborn of 2.8 (P = .04). CONCLUSIONS: Outborn delivery is a significant predictor of mortality for infants with antenatally diagnosed CDH. PMID- 21616233 TI - Ex-utero intrapartum treatment procedure for giant neck masses--fetal and maternal outcomes. AB - BACKGROUND/PURPOSE: For fetuses with giant neck masses and tracheal obstruction, an ex-utero intrapartum treatment (EXIT) procedure allows for safe nonemergent airway management while on placental support. Our objective was to examine fetal and maternal outcomes after EXIT procedure specifically for giant neck masses. METHODS: The medical records of all patients referred to a comprehensive fetal center for a giant neck mass between 2001 and 2010 were reviewed retrospectively. RESULTS: Among 24 patients referred, an EXIT procedure was performed in 12 with evidence of tracheal compression. An EXIT procedure was not performed because of minimal tracheal involvement (n = 8), elective abortion (n = 2), fetal demise (n = 1), or obstetric complication (n = 1). In all fetuses, the airway was successfully secured; tracheal intubation was achieved with rigid bronchoscopy (n = 10), direct laryngoscopy (n = 1), and tracheostomy (n = 1). Eleven patients survived to discharge, whereas 1 patient with significant pulmonary hypoplasia died 8 days after emergency EXIT procedure. Of 11 surviving infants, 10 are neurodevelopmentally intact. All mothers who desired future pregnancies have subsequently had uncomplicated deliveries (n = 6). CONCLUSIONS: Ex-utero intrapartum treatment procedure for giant neck mass can be performed safely for both mother and child. Most fetuses can be orotracheally intubated with minimal long-term morbidity. The potential for future pregnancies is preserved. PMID- 21616234 TI - Pediatric thyroidectomy: a collaborative surgical approach. AB - INTRODUCTION: We have developed a collaborative approach to pediatric thyroid surgery, with operations performed at a children's hospital by a pediatric surgeon and an endocrine surgeon. We hypothesize that this strategy minimizes specialist-specific limitations and optimizes care of children with surgical thyroid disease. METHODS: Data from all partial and total thyroidectomies performed by the pediatric-endocrine surgery team at a tertiary children's hospital between 1995 and 2009 were collected and analyzed retrospectively. Statistical analyses were performed with IBM SPSS software (SPSS, Chicago, IL). RESULTS: Thirty-five children met the inclusion criteria (69% female; median age, 13 years; median follow-up, 1119 days). The indications for operation were thyroid nodule (71%), genetic abnormality with predisposition to thyroid malignancy (17%), multinodular goiter (5.7%), Grave disease (2.9%), and Hashimoto thyroiditis (2.9%). Sixteen children (46%) underwent thyroid lobectomy, and 19 children (54%) underwent total thyroidectomy. Median length of stay was 1 day (1 day after lobectomy vs 2 days after total thyroidectomy, P < .0001). There were 4 cases of transient hypocalcemia after total thyroidectomy, but there were no nerve injuries or other in-hospital complications in either group (overall complication rate, 11%). CONCLUSIONS: For pediatric thyroidectomy and thyroid lobectomy, collaboration of high-volume endocrine and pediatric surgeons as well as pediatric endocrinologists at a dedicated pediatric medical center provides optimal surgical outcomes. PMID- 21616235 TI - A contemporary evaluation of pulmonary function in children undergoing lung resection in infancy. AB - BACKGROUND/PURPOSE: The management of asymptomatic congenital lung lesions is controversial. Some centers recommend resection in infancy, and others prefer observation. Our objective was to evaluate the pulmonary function of children who underwent lung resection at 12 months or younger. We hypothesized that these children would not have a significant reduction in pulmonary function when compared with norms for age. METHODS: All patients at 2 tertiary-care children's hospitals who underwent lung resection at 12 months or younger and are currently older than 5 years were identified and prospectively recruited. Pulmonary function testing was standardized in all patients. RESULTS: Fourteen children were tested prospectively, whereas results were available for another 5 children. Four children were excluded for inability to perform pulmonary function testing (n = 2) or for preexisting pulmonary hypoplasia/syndrome (n = 2). Pulmonary function testing values were considered normal if they were more than 80% of predicted. Forced vital capacity was normal in 14 (93%) of 15 children, and forced expiratory volume in 1 second was normal in 13 (86%) of 15 children. Diffusion capacity and respiratory muscle strength were normal in all children tested. CONCLUSIONS: Most children undergoing lung resection in infancy will have normal pulmonary function tests, supporting our philosophy of early, elective resection of congenital lung lesions. PMID- 21616236 TI - Lung nodules in pediatric oncology patients: a prediction rule for when to biopsy. AB - PURPOSE: The purpose of the study was to develop a prediction rule regarding the factors that most accurately predict the diagnosis of a malignancy in a lung nodule in the pediatric oncology patient. METHODS: A retrospective review of pediatric oncology patients that underwent lung nodule resection between 1998 and 2007 was performed. Multivariable logistic regression was used to create a prediction rule. RESULTS: Fifty pediatric oncology patients underwent 21 thoracotomies and 48 thoracoscopies to resect discrete lung nodules seen on computed tomographic scans during workup for metastasis or routine surveillance. The mean nodule size was 10.43 +/- 7.08 mm. The most significant predictors of malignancy in a nodule were peripheral location (odds ratio [OR], 9.1); size between 5 and 10 mm (OR, 2.78); location within the right lower lobe (OR, 2.43); and patients with osteosarcoma (OR, 10.8), Ewing sarcoma (OR, 3.05), or hepatocellular carcinoma (OR, 2.38). CONCLUSIONS: Lesions that are between 5 and 10 mm in size and peripherally located in patients with osteosarcoma, Ewing sarcoma, or hepatocellular carcinoma are most likely to be malignant. Use of a prediction rule can help guide clinical practice by determining which patients should undergo surgical resection of lung nodules and which patients may be closely observed with continued radiologic studies. PMID- 21616237 TI - Endoscopic management of congenital esophageal stenosis. AB - BACKGROUND/PURPOSE: Congenital esophageal stenosis (CES) is a rare malformation. Endoscopic dilations represent a therapeutic option. This study retrospectively evaluated the efficacy and safety of a conservative treatment of CES. PATIENTS AND METHODS: Patients diagnosed with CES since 1980 by a barium study or endoscopy were reviewed. Endoscopic ultrasonography (Olympus UM-3R-20-MHz radial miniprobe, Olympus Corporation, Tokyo, Japan), available from 2001, allowed for the differential diagnosis of tracheobronchial remnants (TBR) and fibromuscular hypertrophy (FMH) CES. All children underwent conservative treatment by endoscopic dilations (hydrostatic and Savary). RESULTS: Forty-seven patients (20 men) had CES. Fifteen were associated with esophageal atresia; and 8, with Down syndrome. Mean age at the diagnosis was 28.3 months (range, 1 day to 146 months). Symptoms were solid food refusal, regurgitation, vomiting, and dysphagia. Congenital esophageal stenosis was located in the distal esophagus. Endoscopic ultrasonography demonstrated TBR and FMH in 6 patients. One hundred forty-eight dilations in 47 patients were performed. The stenosis healed in 45 (95.7%). Complications were 5 (10.6%) esophageal perforations, hydrostatic (3/32, or 9.3%), and Savary (2/116, or 1.7%). At follow-up, 1 patient with FMH CES and 1 patient with TBR CES required operation for persistent dysphagia. CONCLUSIONS: The conservative treatment yielded positive outcomes in CES. Endoscopic ultrasonography allows for a correct diagnosis of TBR/FMH CES. A surgical approach should be reserved for CES not responsive to dilations. PMID- 21616238 TI - Esophageal stenosis in epidermolysis bullosum: a challenge for the endoscopist. AB - BACKGROUND/PURPOSE: Esophageal stenosis is a severe complication in dystrophic epidermolysis bullosa (EB). Endoscopic dilations may cause mucosal injury with stricture recurrence. Our aim was to describe our referral EB-center experience on safety and long-term efficacy of fluoroscopically guided balloon dilation without endoscopy. METHODS: Over 14 years, 34 patients with EB, previously evaluated with barium esophagogram for dysphagia, underwent balloon esophageal dilation. Under fluoroscopy, a guide wire was introduced via a nostril into the stomach. A 12-mm pneumatic balloon, which passed over the wire, was filled using radio-opaque contrast, dilating the stricture. Orotracheal intubation was avoided. Antibiotics, dexamethasone, and proton-pump inhibitors were administered. Study approval was obtained from our ethical board. RESULTS: Ninety three dilations were performed. Seventeen patients had a single stenosis. The mean age of onset was 18 years (range, 3-47 years). Thirteen patients underwent one dilation. In 6 cases, endoscopy was necessary to visualize the esophageal lumen. Complications included cervical esophageal perforation (2) and transitory dysphagia (10). Thirty patients were feeding within 24 hours. During the follow up, 2 patients required a gastrostomy, and 2 patients underwent fundoplication for gastroesophageal reflux disease. CONCLUSIONS: Fluoroscopically guided balloon dilation in EB is a safe and well-tolerated procedure. An experienced endoscopy team is necessary in certain cases. PMID- 21616239 TI - Custom dynamic stent for esophageal strictures in children. AB - BACKGROUND: Esophageal stenting represents a new strategy to avoid multiple dilations owing to stenosis relapse. Our custom stent improves esophageal motility unlike the widespread self-expandable plastic esophageal stents. The aim of the study was to confirm the efficacy of treatment with silicone custom stents in esophageal stenosis (ES) in pediatric patients. METHODS: A silicone stent of 7 , 9-, or 12.7-mm external diameter is built coaxially on a nasogastric tube that guarantees the correct position. The 2 ends are tailored to allow food passage between stent and esophageal wall. All patients received dexamethasone (2 mg/kg per day) for 3 days and ranitidine/proton-pump inhibitors. Study approval was obtained from our ethical board. RESULTS: From 1988 to 2010, 79 patients with ES, mean age 35.4 months (3-125 months), underwent esophageal hydrostatic/Savary dilations and custom-stent placement, left in place for at least 40 days. Stenting was effective in 70 (88.6%) of 79 patients. Fifty percent of the patients with effective treatment received only one dilation for stent placement. Fourteen patients received more stents successfully. There was one stent-related major complication. CONCLUSION: Our custom stent improves treatment in ES. In caustic injuries, ES stenting represents the first option. In postsurgical ES, we stent after at least 5 dilations. PMID- 21616240 TI - Complete vs partial fundoplication in children with esophageal atresia. AB - PURPOSE: The purpose of the study was to compare outcomes after partial vs complete fundoplication in patients with prior esophageal atresia repair. METHODS: All patients undergoing fundoplication following esophageal atresia repair at a tertiary care pediatric hospital from 1987 to 2006 were retrospectively reviewed. All children had at least 1 year of follow-up postfundoplication. RESULTS: Of 47 children, 31 (66%) had a partial fundoplication and 16 (34%) had complete fundoplication. Demographics, presence of tracheoesophageal fistula, early complications of esophageal atresia repair, gastroesophageal reflux symptoms before fundoplication, and operative details of fundoplication were statistically similar between groups, except for the frequency of hiatus repair during fundoplication (23% vs 69%, P = .004). Patients were followed for a median of 4.98 years (range, 1-17.8 years). Postfundoplication symptoms of vomiting (39% vs 31%), dysphagia (45% vs 38%), retching (10% vs 25%), abnormal findings on barium study, and need for reoperation (19% vs 13%) were not statistically different between groups. However, a greater proportion of children undergoing partial fundoplication achieved long-term symptom- and medication-free recovery (52% vs 13%, P = .012). CONCLUSIONS: Our data suggest that partial fundoplication is associated with a greater likelihood of symptom- and medication-free recovery than complete fundoplication in children with previously repaired esophageal atresia. PMID- 21616241 TI - Outcomes in children with hypoplastic left heart syndrome undergoing open fundoplication. AB - BACKGROUND/PURPOSE: Some institutions recommend early fundoplication in patients with hypoplastic left heart syndrome (HLHS) with signs of gastroesophageal reflux disease because of the risk of reflux-related cardiac events. However, their cardiac physiology may impose high perioperative morbidity and mortality. Therefore, we reviewed our experience with fundoplication in this population to allow for assessment of the risk-benefit ratio. METHODS: A retrospective review of patients with a diagnosis of HLHS who underwent a fundoplication from January 1990 to July 7, 2009, was performed. All patients underwent open fundoplication between first and second stages of cardiac repair. RESULTS: Thirty-nine patients were identified. There were 3 intraoperative complications: hemodynamic instability (n = 2) and a pulmonary hypertensive crisis requiring extracorporeal membrane oxygenation and termination of the procedure (n = 1). There were 27 postoperative complications in 16 patients. There were 2 deaths (4%) within 30 days, and there were 9 deaths (23%) in patients between their first and second stage of cardiac repair during the study period. CONCLUSIONS: Noncardiac surgical procedures in patients palliated for HLHS have a high morbidity and mortality. We recommend that routine fundoplication in this population should only be performed under prospective protocols until the relative risk of operation vs risk of reflux is delineated. PMID- 21616242 TI - Delayed gastric emptying and typical scintigraphic gastric curves in children with gastroesophageal reflux disease: could pyloromyotomy improve this condition? AB - BACKGROUND/PURPOSES: Delayed gastric emptying (DGE) is a cofactor in the etiopathogenesis of gastroesophageal reflux disease (GERD). Scintigraphy is the criterion standard to evaluate gastric emptying (GE). This study aims to define typical scintigraphic activity-time curves (ATCs) related to DGE and esophageal atresia (EA) and to demonstrate the effectiveness of pyloromyotomy (P) in improving GE. METHODS: Since 2002, 83 children underwent Nissen fundoplication. Patients were divided into 2 groups: group I, GERD-only patients; group II, patients with GERD owing to EA. Depending on preoperative scintigraphy, each group was subdivided into 2 subgroups. Before surgery and 1 year after, endoscopy and scintigraphy were performed. In the presence of DGE, P was associated with Nissen fundoplication. Gastric emptying differences at baseline and at follow-up were estimated by the Student t test. Pre- and post-ATCs were evaluated by the chi(2) test. RESULTS: During follow-up, GE completely normalized in subgroups with DGE. Scintigraphic ATC analysis documented an association between DGE and a typical rectilinear fitting, with a higher rate in EA patients. After P, the scintigraphic pattern changed in an exponential manner related to a faster GE. CONCLUSIONS: Delayed gastric emptying is frequent in EA, and the scintigraphic ATCs are typical. Pyloromyotomy is a safe and effective technique to fully normalize GE. PMID- 21616243 TI - Open transumbilical pyloromyotomy: is it more painful than the laparoscopic approach? AB - BACKGROUND: Open transumbilical pyloromyotomy (UMBP) and laparoscopic pyloromyotomy (LAP) have been compared on different outcomes, but postoperative pain as a primary end point had never been assessed. The aim of this study was to compare the use of analgesia in UMBP and LAP patients. METHODS: Infants with hypertrophic pyloric stenosis treated by UMBP in 2008-2009 were matched with LAP treated infants. Demographics, type and use of analgesia, and length of stay were recorded. Statistical analysis was performed using the Fisher exact test. RESULTS: Each group contained 19 patients (N = 38) with comparable demographics and no comorbid condition. Bupivacaine was injected intraoperatively in all UMBP and 89% of LAP infants. There was a trend toward increased acetaminophen use in LAP infants (79% vs 58%, P = .61) in the recovery room. There was no difference in opiates use (3 UMBP vs 1 LAP, P = .60). In the ward, more UMBP patients received acetaminophen (78% vs 53%, P = .03). This difference was significant. Mean postoperative length of stay was similar in both groups. CONCLUSION: Our study suggests that UMBP infants might experience more postoperative pain in the ward, without any impact on various outcomes. A prospective study with a larger sample size should be undertaken to verify these findings. PMID- 21616244 TI - Surgery or endoscopy to treat duodenal duplications in children. AB - BACKGROUND/PURPOSE: Gastrointestinal duplications (duodenal duplications [DDs]) are a rare congenital malformation generally located in or adjacent to the medial border of the duodenal wall. The goal of therapy is surgical excision. Conservative endoscopic management represents an alternative option. AIM: The aim of the study was to highlight the role of endoscopic ultrasound (EUS) in guiding the endoscopic or surgical treatment of DD. METHODS: Between 2002 and 2010, 6 patients (2 male; mean age, 7.83 years; range, 2-18 years), all with recurrent acute pancreatitis, were diagnosed with DD by ultrasound and magnetic resonance imaging. Endoscopy was always performed together with EUS (Olympus UM-3R 20-MHz radial miniprobe, Tokyo, Japan). An endoscopic section of the common duodenal-DD wall, using a precut needle or sphincterotome, was chosen by EUS when the biliary tree was not involved in the DD. Otherwise, surgery with duodenotomy and complete opening of the common wall was used. RESULTS: After EUS evaluation, endoscopic treatment was successfully performed in 4 patients, 2 of whom required surgical treatment. Bleeding occurred in 1 patient after endoscopic resection and in 1 patient after surgery. The mean follow-up time without pathologic signs was 3.3 years (range, 0.25-8). CONCLUSIONS: Endoscopic ultrasound can effectively guide surgical or endoscopic therapies. Bleeding is a possible complication. PMID- 21616245 TI - Pathologic changes in biliary dyskinesia. AB - PURPOSE: For children with upper abdominal pain and evaluation for acalculous biliary disease, laparoscopic cholecystectomy is an accepted treatment with inconsistent outcomes. The purpose of this study was to identify predictors of outcomes. METHODS: One hundred sixty-seven children underwent laparoscopic cholecystectomy at a single children's hospital. Radiographic findings, histopathology, family history, and demographics (sex, age, height, weight, body mass index-for-age percentile) were evaluated as predictors of postoperative symptomatic resolution using a binomial probability model. The data for radiologic studies and pathologic specimens were obtained via re-review in a blinded fashion. RESULTS: Of 167 children, 43 (25.7%) had a preoperative diagnosis of biliary dyskinesia and 41 (95.3%) had documented follow-up. Mean follow-up was 8.4 months. Twenty-eight patients (68.3%) had symptom resolution. Ejection fraction less than or equal to 15%, pain upon cholecystokinin injection, and a family history of biliary disease were not predictors of symptomatic resolution. Nonoverweight patients (body mass index-for-age <85th percentile) were more likely to have symptom resolution than their overweight counterparts (odds ratio, 2.13). Most patients (68.3%) had a pathologic gallbladder on blinded review. However, this did not correlate with outcome. CONCLUSIONS: Most gallbladders removed for biliary dyskinesia are pathologic. Being overweight can be considered a relative contraindication to cholecystectomy for biliary dyskinesia. PMID- 21616246 TI - Parenteral aluminum induces liver injury in a newborn piglet model. AB - PURPOSE: Parenteral nutrition-associated cholestasis remains a significant problem, especially for the surgical neonate. Aluminum is a toxic element known to contaminate parenteral nutrition. We hypothesize that parenterally administered aluminum causes liver injury similar to that seen in parenteral nutrition-associated cholestasis. METHODS: Twenty 3- to 6-day-old domestic pigs were divided into 5 equal groups. A control group received daily intravenous 0.9% sodium chloride. Each subject in experimental groups received intravenous aluminum chloride at 1500 MUg/kg per day for 1, 2, 3, or 4 weeks. At the end of the study, blood was sampled for direct bilirubin and total bile acid levels. Liver, bile, and urine were sampled for aluminum content. Liver tissue was imaged by transmission electron microscopy for ultrastructural changes. RESULTS: Transmission electron microscopy revealed marked blunting of bile canaliculi microvilli in all experimental subjects but not the controls. Serum total bile acids correlated with the duration of aluminum exposure. The hepatic aluminum concentration correlated with the duration of aluminum exposure. CONCLUSIONS: Parenterally infused aluminum resulted in liver injury as demonstrated by elevated bile acids and by blunting of the bile canaliculi microvilli. These findings are similar to those reported in early parenteral nutrition-associated liver disease. PMID- 21616247 TI - Hepatobiliary anomalies in conjoined twins. AB - INTRODUCTION: Conjoined twinning is an extremely rare anomaly. Very few diagrammatic descriptions are provided for the various hepatobiliary anomalies seen in these twins. We aimed to review our experience with the various subtypes of hepatobiliary anomalies and their association with the inability to separate as well as provide diagrammatic descriptions of these anomalies. METHODS: We retrospectively reviewed our experiences within separating twins. We reviewed patterns of hepatobiliary anomalies and the required investigations and intraoperative workups. RESULTS: Of the 60 cases we evaluated, 28 were successfully separated. The reasons for nonseparation were possession of: a single heart, major communicating hearts, or major chromosomal anomalies. The liver was involved in 17 cases (60.7%) in the operative group and 23 cases (71.8%) in the nonoperative group. All cases had a computed tomographic scan and ultrasound as preoperative workup. Only 2 cases required a magnetic resonance cholangiopancreatography for preoperative evaluation. Intraoperative ultrasound was not used, and only 3 cases required an intraoperative cholangiogram. Diagrammatic depictions of the various categories of anomalies are presented. CONCLUSION: In our experience, we did not find hepatobiliary anomalies to be the sole reason for inseparability in any of the conjoined sets. Hepatobiliary anomalies seem to be more frequent in the nonseparable group. PMID- 21616248 TI - National trends in the surgical management of Meckel's diverticulum. AB - BACKGROUND: Treatment recommendations for Meckel's diverticulum (MD) come mostly from single-institution case series. The objective of this study was to review the surgical management and outcomes of children undergoing Meckel's diverticulectomy using contemporary data from a national database. METHODS: We queried 2007 to 2008 data from the Pediatric Health Information System database and analyzed demographic and outcome variables for patients undergoing surgical resection of MD. Cases were classified as primary (symptomatic MD) or secondary (incidental MD). Outcomes in primary cases were compared between open and laparoscopic approaches. Statistical analyses were performed using SPSS (Chicago, IL). RESULTS: Eight hundred fifteen children underwent Meckel's diverticulectomy. Meckel's diverticulectomy was more common in boys (boy-girl, 2.3:1), and half (53%) of the children required surgery before their fourth birthday. More cases (n = 485; 60%) were classified as primary, and most children were approached by laparotomy (75%). The most common presentations for primary cases were obstruction (30%), bleeding (27%), and intussusception (19%). In the primary group, patients treated with the laparoscopic approach had a shorter length of stay (open approach, 5.7 +/- 5.2 days; laparoscopic approach, 4.3 +/- 2.7 days; P < .02). CONCLUSION: These data describe current trends in the surgical treatment of MD in the United States. Laparoscopic Meckel's diverticulectomy appears to shorten length of stay but is used much less frequently than the traditional open approach. PMID- 21616249 TI - Development and validation of a pediatric laparoscopic surgery simulator. AB - BACKGROUND: Although a validated simulator exists for adult laparoscopy, there is no pediatric counterpart. The objective of this study is to develop and validate a pediatric laparoscopic surgery (PLS) simulator. METHODS: A PLS simulator was developed. Participants were stratified according to level of expertise and tested on the fundamentals of laparoscopic surgery (FLS) and PLS simulators. A subsequent group was tested exclusively on the PLS simulator. RESULTS: The PLS intracorporeal suturing score was lower than its adult counterpart (P = .02). The PLS pattern-cutting score was higher than in the FLS simulator (P < .001). If the latter was eliminated from the calculation, the revised total FLS score was significantly better than the revised PLS score. When all participants were combined, total PLS scores as well as performance on 3 of 5 tasks allowed differentiation between novice, intermediate, and expert. CONCLUSIONS: The PLS simulator was able to discriminate between the novice, intermediate, and expert using the total PLS score and the performance on 3 of the 5 tasks, thus providing evidence for construct validity. The other 2 tasks will require formal modification or a change in the scoring metrics to establish their independent construct validity. PMID- 21616250 TI - Single-incision laparoscopic surgery in children: initial single-center experience. AB - BACKGROUND: In continued efforts to further improve the advantages of minimally invasive surgery to patients, surgeons have developed single-incision laparoscopic techniques. We report our initial experience in children with a variety of single-site procedures. METHOD: A retrospective chart review was performed on patients who underwent a single-site procedure from April 2009 to April 2010. RESULTS: There were 142 consecutive procedures: 24 cholecystectomies, 103 appendectomies for nonperforated appendicitis, 2 splenectomies, 1 combined splenectomy/cholecystectomy, 8 ileocecectomies, 2 Meckel diverticulectomies, 1 small bowel duplication resection, and 1 jejunal stricture resection. There were 12 conversions to conventional laparoscopy: 10 during appendectomy and 2 during cholecystectomy. Mean operative time was 34 minutes for appendectomy, 73 minutes for cholecystectomy, 90 minutes for splenectomy, 116 minutes for combined splenectomy/cholecystectomy, 86 minutes for ileocecectomy, and 43 minutes for the small bowel procedures. The only complications were umbilical surgical site infections after appendectomy in 6 patients. CONCLUSION: This institution's preliminary experience suggests that single-incision laparoscopic surgery in children has at least comparable outcomes to conventional laparoscopic surgery. However, prospective data are needed to prove that single-incision laparoscopic surgery is superior to conventional laparoscopy. PMID- 21616251 TI - Prematurity, not age at operation or incarceration, impacts complication rates of inguinal hernia repair. AB - PURPOSE: Inguinal hernia repair (IHR) remains the most common procedure in pediatric surgery. Although postoperative sequelae are well described, we examined if prematurity and age were important determinants of postoperative complications. METHODS: A retrospective review of children younger than 2 years undergoing IHR from 2004 to 2007 was performed, with a minimum of 1-year follow up. Patients were segregated into groups based on age at diagnosis (A, 0-3; B: 4 26; C: 27-52; D: 53-104 weeks), with or without prematurity. Incarceration rates were investigated. Complications were categorized as major (vas injury, recurrence, testicular atrophy) or minor (wound infection, "high" testicle, hydrocele) and compared. RESULTS: Two hundred sixty-eight patients were analyzed (98 premature), with 14 major complications (5.2%) and 26 (9.7%) minor complications overall. Groups A and B accounted for more major (12/14) and minor complications (22/26) when compared with groups C and D (P < .005). In patients less than 26 weeks (groups A and B), premature infants had more complications than term infants (27.7% vs 12.1%, P = .01). Of 22 patients with incarcerated hernias, 2 (9.1%) had major complications (P < .5 vs nonincarcerated patients). CONCLUSIONS: Our study suggests that prematurity, rather than age at operation or incarceration, affects complication rates after IHR. This information should be used to frame the discussion of informed consent for this commonly performed procedure. PMID- 21616252 TI - Sclerotherapy for lymphatic malformations in children: a scoping review. AB - PURPOSE: This scoping review assesses the literature and summarizes the current evidence on sclerotherapy for the treatment of lymphatic malformations in pediatric patients. METHODS: A comprehensive search of published and unpublished literature was conducted using multiple databases. Title, abstract, and full-text screening was conducted by 2 independent clinicians. All discrepancies were resolved during consensus meetings. RESULTS: A total of 182 articles were retrieved. Forty-four articles were removed as duplicates, and 11 articles were added after reviewing prominent studies. After full-text abstraction, 44 articles and 2 conference proceedings (N = 882 patients) were included in the final results. Twelve articles were classified as level II and 34 articles as level IV evidence. Picibanil (OK-432) was the primary agent used in most included studies. Postinjection symptoms with OK-432 were primarily fever, swelling, and erythema at the site. Life-threatening complications were uncommon and involved postinjection swelling of cervical lesions causing airway compromise. CONCLUSIONS: The literature regarding sclerotherapy for lymphatic malformations is of a low level of evidence and suffers from a lack of standardization. Randomized clinical trials focused on OK-432, bleomycin, or alcoholic solution of zein; standardized dosing protocols; and consistent and reliable outcome reporting will be necessary for further development of treatment guidelines. PMID- 21616253 TI - Utility of amylase and lipase as predictors of grade of injury or outcomes in pediatric patients with pancreatic trauma. AB - INTRODUCTION: Grade of injury, serum amylase, and lipase are markers used to assess pancreatic injury. It is unclear how amylase and lipase relate to grade of injury or predict outcome. We hypothesize that serum amylase and lipase are good predictors of grade of injury and outcomes in patients with pancreatic trauma. METHODS: This study is a multicenter review from 9 pediatric trauma centers of all children admitted to their institution over 5 years with a pancreatic injury. Initial as well as peak amylase and lipase values were analyzed with relation to pancreatic grade, length of stay, and outcomes. RESULTS: One hundred thirty-one records were analyzed. There were 44 girls and 85 boys with an average age of 9.0 +/- 0.4 years. The mean injury severity score (ISS) score was 15.5 +/- 1.2 SE. The average length of stay (in days) was analyzed by grades 0 (3.93), 1 (7.73), 2 (13.4), 3 (18.4), 4 (31), and 5 (13.5). Neither initial nor peak amylase/lipase correlated with grade of injury. Neither amylase nor lipase predicted length of stay or mortality. Maximal amylase was highly predictive of developing a pseudocyst. CONCLUSION: There seems to be limited value for repetitive routine amylase and lipase levels in the management of pediatric trauma patients with pancreatic injury. PMID- 21616254 TI - Pediatric firearm injuries: a 10-year single-center experience of 194 patients. AB - BACKGROUND: The objective was to investigate the relationship of high gun ownership and gun death rate on children and determine predictors influencing the incidence and outcome of pediatric firearm injuries in a major pediatric level 1 trauma center. METHODS: We performed a retrospective review of our trauma registry to identify hospital admissions between April 1999 and March 2010. We extracted demographic and geographic data, seasonal variation, injury type, firearm type, and outcome. RESULTS: We identified 194 firearm injuries. The incidence did not change during the past decade. Most occurred during the second half of the year (61.4%). Mean age was 12.2 +/- 4.6 years (range, 0.4-19.2 years). Unintentional shootings accounted for 100 injuries followed by assaults (n = 55) and innocent bystanders (n = 39). African American children were most often injured because of a violent cause (60.3%), whereas white children were shot unintentionally (80.1%). Powder-propelled firearms caused 82.5% of injuries. Overall, 17.5% of children required an operation, and 9.3% died. CONCLUSIONS: The overwhelming majority of children were injured after a gun went off unintentionally, whereas most African American children were shot violently. We identified certain seasonal and geographic clusters. These data can be used to target gun injury prevention programs. PMID- 21616255 TI - The significance of pseudoaneurysms in the nonoperative management of pediatric blunt splenic trauma. AB - PURPOSE: Nonoperative management is the standard of care for hemodynamically stable pediatric and adult blunt splenic injuries. In adults, most centers follow a well-defined protocol involving repeated imaging at 24 to 48 hours, with embolization of splenic pseudoaneurysms (SAPs). In children, the significance of radiologically detected SAP has yet to be clarified. METHODS: A systematic review of the medical literature was conducted to analyze the outcomes of documented posttraumatic SAP in the pediatric population. RESULTS: Sixteen articles, including 1 prospective study, 4 retrospective reviews, and 11 case reports were reviewed. Forty-five SAPs were reported. Ninety-six percent of children were reported as stable. Yet, 82% underwent splenectomy, splenorrhaphy, or embolization. The fear of delayed complications owing to SAP was often cited as the reason for intervention in otherwise stable children. Only one child with a documented pseudoaneurysm experienced a delayed splenic rupture while under observation. No deaths were reported. CONCLUSIONS: There is no evidence to support or dispute the routine use of follow-up imaging and embolization of posttraumatic SAP in the pediatric population. At present, the decision to treat SAP in stable children is at the discretion of the treating physician. A prospective study is needed to clarify this issue. PMID- 21616256 TI - Traumatic pseudoaneurysms of the liver and spleen in children: is routine screening warranted? AB - BACKGROUND: Although blunt injury to the spleen and liver can lead to pseudoaneurysm formation, current surgical guidelines do not recommend follow-up imaging. Controversy exists regarding the clinical implications of these traumatic pseudoaneurysms as well as their management. METHODS: Retrospective review of children treated nonoperatively for isolated blunt liver and spleen trauma between 1991 and 2008 was undertaken. Patient demographics, grade of injury, and follow-up Doppler ultrasound results were obtained. RESULTS: Three hundred sixty-two children were identified. One hundred eighty-six of them had splenic injuries, and 10 (5.4%) developed pseudoaneurysms. They were associated with grade III (3/39 [8%]) and grade IV (7/41 [17%]) injuries. In 7 patients, the pseudoaneurysm thrombosed spontaneously. Angiographic embolization was required in 2 children, and one underwent emergency splenectomy for delayed hemorrhage. Of the 176 patients who had liver injuries, 3 (1.7%) developed pseudoaneurysms. All 3 were associated with grade IV injuries (3/11 [27%]). One child underwent early embolization, while 2 developed delayed hemorrhage requiring emergent treatment. CONCLUSIONS: Pseudoaneurysm development after blunt abdominal trauma is associated with high-grade splenic and liver injuries. Routine screening of this group of patients before discharge from hospital may be warranted because of the potential risk of life-threatening hemorrhage. PMID- 21616257 TI - Central venous catheter database: an important issue in quality assurance. AB - BACKGROUND/PURPOSE: The purpose of this study was to analyze the factors that affect the longevity of central venous catheters. METHODS: Comprehensive clinical data recorded during insertion and removal of totally implantable devices (TID) and tunneled lines (TL) from October 1988 to January 2009 were analyzed. Univariate and multivariate Cox proportional hazards regression models were used to identify clinical factors that predict catheter longevity. RESULTS: Information was available for 1167 central venous catheter insertions in 858 patients, 648 TID and 509 TL. Univariate analysis detected longer device longevity in the following: TID longer than TL (P < .0001), catheter tip in the superior vena cava (SVC)/right atrial junction (P < .0001), and right side greater than left (P = .002). Shorter device longevity was observed in lines used for total parenteral nutrition (P < .0001) and young age (P < .0001). Multivariate model detected the following: hazard of removal for TID is 0.304 that of TL (P < .0001) and SVC is 0.525 that of other locations (P = .0005). Hazard decreases by 5.4% for every 1-year increase in patient age (P < .0004). CONCLUSION: Multiple confounding factors were encountered. However, the single most important factor in catheter longevity that is influenced by the surgeon is tip location in the SVC/right atrial junction. PMID- 21616258 TI - Outcome of peripherally inserted central venous catheters in surgical and medical neonates. AB - BACKGROUND/PURPOSE: Peripherally inserted central venous catheters (PICCs) are commonly used for neonatal vascular access. The aim of this study was to look at PICC line complication rates and possible predictors of PICC infection in a neonatal intensive care unit. METHOD: This was a prospective study of 226 neonates who had PICCs on our neonatal intensive care unit between January 2006 and June 2009. Complete data was available on 218 neonates who had 294 PICC lines. Criteria for catheter-related sepsis was positive blood cultures (peripheral/central) and/or a positive catheter tip culture after removal in the presence of a clinical suspicion of line sepsis. RESULTS: Of 218 neonates, 132 (169 lines) were medical, and 86 (125 lines) were surgical. Our PICC line infection rate was 17 infections per 1000 catheter-days. Surgical neonates had infection rates of 24.8% compared with 18.3% of medical neonates (P < .18). The odds ratio for a PICC infection was 3.1 (95% confidence interval, 1.64-5.87) if the catheter was in situ for 9 days or more, P < .01. Coagulase-negative staphylococcus was isolated from 55 (89%) of 62 blood cultures. CONCLUSIONS: Our PICC infection rate was 17 per 1000 catheter-days. The length of catheter stay was the only predictor of PICC infection. PMID- 21616259 TI - Ethanol lock therapy to reduce the incidence of catheter-related bloodstream infections in home parenteral nutrition patients with intestinal failure: preliminary experience. AB - BACKGROUND: Catheter-related bloodstream infections (CRBSI) cause morbidity and mortality in patients with intestinal failure dependent on parenteral nutrition. Ethanol lock of central venous catheters may decrease CRBSI, but limited pediatric data are available. METHODS: Home parenteral nutrition patients with at least one previous CRBSI were initiated on a 70% ethanol lock protocol for a minimum of 4 hours. Infection rates (per 1000 catheter days) before and after initiation of ethanol locks were compared using a paired t test. RESULTS: Ten patients (4 girls; median age, 44 months [range, 31-129 months]) began ethanol lock therapy after a total of 91 CRBSIs (37 gram-positive, 30 gram-negative, and 24 fungal) with a mean of 10.2 +/- 6.2 per 1000 catheter days. Patients received ethanol lock for an average of 227 +/- 64 days with only 3 CRBSI occurring (CRBSI rate of 0.9 +/- 1.8 per 1000 catheter days [P = .005]). Central venous catheter replacements decreased from 5.6 per 1000 days to 0.3 per 1000 days posttherapy (P = .038). Ethanol lock was discontinued in 2 of 10 patients because of catheter thrombosis. CONCLUSION: Preliminary results demonstrate a significant decrease in CRBSI with a 70% ethanol lock protocol. Catheter thrombosis may be a limitation that needs to be addressed. With such a dramatic therapeutic effect, a randomized trial is feasible and should be performed. PMID- 21616260 TI - Success in the Pediatric Surgery Match: a survey of the 2010 applicant pool. AB - BACKGROUND/PURPOSE: Traditionally, basic science research and publication record have led to a successful Pediatric Surgery Match. With changing applicant research backgrounds, we evaluated if these or other factors still apply. METHODS: A SurveyMonkey questionnaire was distributed to 57 applicants with known contact information. We assessed demographic/financial data, application details and match results, research experience, publications, presence of a pediatric surgery fellowship at their home program, and applicant ranking criteria. RESULTS: Forty-three (75%) responses were received. Twenty-five candidates matched, 12 (48%) to 1 of their first 3 choices. The median number of programs applied to was similar for matched and unmatched candidates (30), but matched candidates attended more interviews (21 vs 14.5; P = .03). Matched applicants had more publications (9.5 vs 5.1; P = .03), although research experience was similar to unmatched candidates. Research focus for matched vs total applicants included basic science (5 vs 12), clinical (4 vs 6), and both (11 vs 16). Five candidates matched without research experience. Ten (40%) applicants matched to institutions where they completed residency/research/fellowship training. Twelve (49%) applicants matched from programs without a fellowship program. CONCLUSION: A strong publication record remains important, although clinical research is being valued more. Candidates from nonfellowship programs can be successful. This information may be useful to mentor future applicants and lays the foundation for a critical evaluation of the match process. PMID- 21616261 TI - Lights, camera, surgery: a novel pilot project to engage medical students in the development of pediatric surgical learning resources. AB - BACKGROUND/PURPOSE: It is important to engage junior medical students in the pediatric surgical environment to showcase possible future career choices. Our aim was to assess how the students valued their experience in the realm of clinical learning, exposure to surgical careers, and development of skill sets necessary for creating learning resources. METHODS: A novel pilot project entitled Lights, Camera, Surgery engaged 13 medical students in the production of instructional videos of basic surgical procedures. An electronic survey questionnaire allowed the students to provide formal feedback on the project outcomes. RESULTS: Of the students who responded, 87.5% expressed appreciation of the enhanced clinical experience. All of the students either agreed or strongly agreed that the project afforded them valuable leadership experience, practical skills in creating educational learning resources, and opportunities to explore careers in surgery. All of the students either agreed or strongly agreed that the project allowed them to gain valuable skills in educational video production. The project videos are now available as educational tools. CONCLUSIONS: Engaging medical students in the production of surgical videos potentially improves leadership skills and promotes the use of educational resources while affording them opportunities to explore pediatric surgery as a future career choice. PMID- 21616262 TI - Report of the 42nd annual meeting of the Canadian Association of Pediatric Surgeons. AB - This is a report of the 42nd annual meeting of the Canadian Association of Pediatric Surgeons. PMID- 21616263 TI - Long-term follow-up of a child with primary lymph node gastrinoma and Zollinger Ellison syndrome. AB - A 10-year-old child with Zollinger-Ellison syndrome and primary lymph node gastrinoma is reported to emphasize the difficulties encountered in management and the value of long-term follow-up. The gastrinoma was present in a lymph node close to the greater curvature of the stomach. Primary lymph node gastrinomas are relatively rare in children and, to the best of our knowledge, have not previously been reported at this location. PMID- 21616264 TI - Selective ligation of portal vein and hepatic artery for ruptured hepatocellular carcinoma in a 13-year-old boy. AB - Hepatocellular carcinoma (HCC) in children is rare, and the prognosis has been poor because of its advanced stage at diagnosis and unresponsiveness to chemotherapy. We report a 13-year-old boy with ruptured HCC in the left trisegment. When hemostasis of the ruptured surface was achieved in the emergency operation, the left branch of the portal vein and the left hepatic artery were ligated at the same time. The volume of the future liver remnant (FLR), that is, his right posterior sector, increased from 56% on admission to 70% of his standard liver volume on day 2. Blood level of serum protein induced by vitamin K absence or antagonist II started to decrease immediately. Left trisegmentectomy was successfully performed 10 days later, followed by chemotherapy. He has been well with a 2-year survival without recurrence. When the FLR is considered relatively small for a major hepatic resection, the selective ligation of the portal vein and the hepatic artery, which feed HCC, seems to be beneficial. This is because it may induce enlargement of the FLR, increasing the safety of the hepatectomy as preoperative portal vein embolization does before a major hepatectomy in adult patients with HCC, and the latter suppresses the tumor while waiting for the planned hepatectomy. PMID- 21616265 TI - Laparoscopic nephroureterectomy for Wilms' tumor: oncologic considerations. AB - Wilms' tumor is the most common malignant renal tumor of childhood. Surgical resection is an important aspect of therapy and is traditionally performed through an open transabdominal approach. The advent of advanced laparoscopic techniques for benign renal lesions has led to interest in applying a minimally invasive approach to Wilms' tumors in children. We describe a 2-year-old girl who presented with a right renal mass measuring 18 * 13 cm with peritoneal seeding and pulmonary metastatic disease. Wilms' tumor was confirmed on open biopsy. After neoadjuvant chemotherapy, a laparoscopic resection of the tumor with right radical nephroureterectomy, retroperitoneal lymph node dissection, and resection of peritoneal metastases was achieved using one 12-mm and four 5-mm ports. The specimens were removed in an endoscopic retrieval bag through a small Pfannenstiel incision by extending the 12-mm port site. The patient had an uncomplicated recovery and was discharged on the fifth post-operative day. She remains disease free at 19 months postoperatively. Minimally invasive techniques may be considered for resection of Wilms' tumor provided oncologic principles are carefully followed. PMID- 21616266 TI - Laparoscopic nephron-sparing resection of synchronous Wilms tumors in a case of hyperplastic perilobar nephroblastomatosis. AB - Diffuse hyperplastic perilobar nephroblastomatosis (DHPLN) is a rare precursor lesion of Wilms tumor (WT). Because of the increased risk to develop WT in either kidney, current management algorithms of DHPLN merit nephron-sparing strategies, beginning with chemotherapy and close radiographic monitoring into late childhood. After resolution of DHPLN, subsequent detection of a renal nodule mandates resection to exclude WT. Here, we report the case of a 4-year-old girl who developed 2 synchronous nodules in the right kidney more than 2 years after completion of therapy for DHPLN. Because of the early detection and peripheral location of these 2 nodules, laparoscopic nephron-sparing resection of each was performed using ultrasonic dissection. Both nodules were determined on pathology to be favorable histology WT with negative surgical margins. The child was placed on vincristine and actinomycin D therapy for 18 weeks. PMID- 21616267 TI - Resection of a duodenal web using single-incision pediatric endosurgery. AB - Single-incision pediatric endosurgery (SIPES) is used in many centers for routine cases such as appendectomies and cholecystectomies, but more complex procedures are still infrequently performed. We report the case of a 9-month-old girl with Down syndrome diagnosed with a duodenal web who underwent duodenal web resection and tapering using a SIPES technique. The procedure was performed through a single 2-cm incision in the umbilicus and took 209 minutes. Postoperatively, the patient was feeding well, gaining weight, and had no appreciable scar at a follow up of 6 months. SIPES duodenal web resection in an infant is a reasonable alternative to conventional triangulated laparoscopy that can be performed safely with good functional and cosmetic postoperative results. PMID- 21616268 TI - Preoperative administration of Sudan III and successful treatment of persistent chylous ascites in a neonate. AB - Congenital chylous ascites is a rare entity, and surgical treatment is confined to selected intractable cases. We report 2 cases of refractory congenital chylous ascites successfully treated with preoperative administration of lipophilic dye (Sudan III) followed by abdominal systematic surgical exploration, cauterization, and fibrin glue application. PMID- 21616269 TI - A case report of bilateral cervical chondrocutaneous remnants with review of the literature. AB - Cervical chondrocutaneous remnants are very rare entities. They are thought to originate either from the second branchial arch or from auricular tissues. To date, less than 40 cases have been reported in the medical literature, and only 7 cases were bilateral. We report the case of a 1-month-old girl presenting with bilateral neck lesions since birth, with no other anomalies. Complete surgical excision was performed, the pathology of which confirmed the diagnosis of chondrocutaneous remnants. Follow-up after 9 months showed no clinical evidence of complications or recurrence. PMID- 21616270 TI - Ovarian fibroma with marked ascites and elevated serum levels of CA-125 in a young girl. AB - We report a case of ovarian fibroma with marked ascites and elevated serum CA-125 levels in a young girl. Ovarian fibromas are rare in children. They usually present as a solid mass and may be associated with ascites and elevated serum CA 125 levels. Because of their solid nature and these associations, they can be mistaken for a malignant tumor, resulting in unnecessary oophorectomy. Ovarian fibromas are benign neoplasms, and the prognosis is extremely good. Surgical management should be an ovarian-sparing tumor excision. Although uncommon in pediatric patients, ovarian fibromas should be included in the differential diagnosis of ovarian mass in children. PMID- 21616271 TI - Novel titanium constructs for chest wall reconstruction in children. AB - PURPOSE: We have previously reported the use of the vertical expandable prosthetic titanium rib (VEPTR) for treatment of thoracic dystrophy. This report describes our experience with this device and other novel titanium constructs for chest wall reconstruction. METHODS: This is a retrospective chart review of all children and adolescents undergoing chest wall reconstruction with titanium constructs between December 2005 and May 2010. RESULTS: Six patients have undergone chest wall reconstruction with VEPTR or other titanium constructs. Four had chest wall resection for primary malignancy, 1 had metastatic chest wall tumor resection, and 1 had congenital chest wall deformity. There were no immediate complications, and all patients have exhibited excellent respiratory function with no scoliosis. CONCLUSIONS: Chest wall reconstruction after tumor resection or for primary chest wall deformities can be effectively accomplished with VEPTR and other customized titanium constructs. Goals should be durable protection of intrathoracic organs and preservation of thoracic volume and function throughout growth. Careful preoperative evaluation and patient-specific planning are important aspects of successful reconstruction. PMID- 21616272 TI - Ten-year review of groin laparoscopy in 1001 pediatric patients with clinical unilateral inguinal hernia: an improved technique with transhernia multiple channel scope. AB - PURPOSE: The purpose of this 10-year review of data is to verify the effectiveness, safety, and appropriate age group for using a multichannel scope during inguinal herniorrhaphy in pediatric patients with clinical unilateral inguinal hernia to evaluate for contralateral patent processus vaginalis (CPPV). METHODS: The data evaluated are age, sex, negative findings, positive findings, false positives, false negatives, recurrences, date of recurrence, and complications. Patients who clinically had bilateral hernias or were born prematurely were excluded. A 5-mm, 30 degrees multichannel scope was used through the ipsilateral open hernia sac to explore the contralateral internal ring. A Fogarty catheter was used through one of the channels of the scope to probe the contralateral side in instances of questionable patent processus vaginalis. RESULTS: One thousand one patients were studied, and a total of 237 CPPVs (23%) were identified. The highest incidence of CPPV was found in those patients younger than 1 year (44%). Contralateral patent processus vaginalis was identified and ligated in 34% of patients younger than 2 years, 20% in patients 2 to 8 years old, and 17% of patients 9 to 18 years old. There were no false positives and 6 false negatives (0.6%) of the contralateral side. There were 3 (0.3%) recurrent inguinal hernias of the ipsilateral side and no complications. CONCLUSIONS: The use of a multichannel scope through the ipsilateral open hernia sac during inguinal herniorrhaphy in pediatric patients with clinical unilateral inguinal hernia to evaluate for CPPV proved to be effective, cost-effective, and safe. Our procedure eliminated any additional scars and the cost of trocars and permitted us to probe the contralateral internal ring. Unnecessary open exploration was spared in 56% of children younger than 1 year and proved to be useful in all pediatric patients up to the age of 18 years. PMID- 21616273 TI - Massive hydrops of the gallbladder mimicking a choledochal cyst. PMID- 21616275 TI - Letter to the Editor regarding comparison of anorectal angle and continence after Georgeson and Pena procedures for high/intermediate imperforate anus. PMID- 21616276 TI - Nonneoplastic cystic lesions of pancreas: a practical clinical, histologic, and radiologic approach. AB - Cystic lesions of pancreas are increasingly identified due to widespread use and improved resolution of cross-sectional imaging. These lesions are broadly classified into neoplastic and nonneoplastic cysts. Nonneoplastic cysts have no malignant potential and include pseudocysts, retention cysts, benign epithelial cysts, abscesses, duodenal wall cysts (diverticula), lymphoepithelial cysts, and the recently described mucinous nonneoplastic cyst. Herein, we report the clinical presentation, histology, and imaging findings of nonneoplastic cystic lesions of the pancreas together with pancreatic necrosis. PMID- 21616277 TI - Myelography: a primer. AB - Despite recent trends toward evaluation of back and neck pain with magnetic resonance imaging, myelography and postmyelography computed tomography continue to play an important role in the workup of many patients with spinal pathology. We present techniques for the safe and efficient performance of myelography, lumbar, and cervical puncture, which remain important skills to be mastered by radiology residents and fellows. PMID- 21616278 TI - Magnetic resonance imaging of cardiac tumors: part 1, sequences, protocols, and benign tumors. AB - Cardiovascular magnetic resonance imaging (CMRI) is the reference noninvasive technique for assessment and characterization of a suspected cardiac or juxta cardiac mass. The multiplanar assessment of anatomy, tissue composition, and functional impact afforded by CMRI allows for early differentiation between a nonneoplastic mass and a tumor mass, be it benign or malignant. CMRI has superior tissue contrast resolution compared with competing noninvasive imaging modalities (echocardiography and multidetector computed tomography). A number of different imaging sequences are employed for a comprehensive CMRI assessment. Black-blood prepared sequences are mainly used for tissue characterization and assessment of lesion enhancement characteristics. Bright blood prepared sequences provide functional information, such as lesion mobility and impact on adjacent valves and chambers. Atrial myxoma is the most common primary benign cardiac tumor and can present with obstructive or embolic phenomena. Fibroelastomas are smaller lesions that do not alter hemodynamic function but can cause symptoms secondary to systemic embolization. Less common benign cardiac tumors include lipoma, fibroma, hemangioma, paraganglioma, and cystic tumor of the atrioventricular nodal region. PMID- 21616279 TI - Magnetic resonance imaging of cardiac tumors: part 2, malignant tumors and tumor like conditions. AB - Cardiovascular magnetic resonance imaging (CMRI) is the reference noninvasive imaging technique for assessment and characterization of a suspected cardiac or juxta-cardiac mass. The multiplanar assessment of anatomy, tissue composition, and functional impact afforded by CMRI allows for early differentiation between a nonneoplastic mass and a tumor mass, be it benign or malignant. Malignant cardiac tumors have a poor prognosis; however, early detection and characterization confer some survival advantage, enabling early instigation of chemotherapy and/or consideration of a surgical debulking procedure. Cardiac metastases are far more common than primary tumors and are an important consideration in patients with disseminated disease. Angiosarcoma accounts for the majority of primary malignant lesions. Less common primary malignant cardiac tumors include sarcomas with myofibroblastic differentiation, lymphoma, rhabdomyosarcoma, pericardial mesothelioma, and pericardial synovial sarcoma. A number of benign masses and normal anatomical variants can cause confusion to the inexperienced observer and must be recognized to avoid unnecessary intervention. These include intracardiac thrombus, bronchogenic and pericardial cysts, and anatomical structures, such as the Crista terminalis and moderator band. PMID- 21616280 TI - Bland-White-Garland syndrome in an elderly woman. PMID- 21616282 TI - The impact of patient and lesion complexity on clinical and angiographic outcomes after revascularization with zotarolimus- and everolimus-eluting stents: a substudy of the RESOLUTE All Comers Trial (a randomized comparison of a zotarolimus-eluting stent with an everolimus-eluting stent for percutaneous coronary intervention). AB - OBJECTIVES: The aim of this study was to investigate the impact of patient and lesion complexity on outcomes with newer-generation zotarolimus-eluting stents (ZES) and everolimus-eluting stents (EES). BACKGROUND: Clinical and angiographic outcomes of newer-generation stents have not been described among complex patients. METHODS: Patients enrolled in the RESOLUTE All Comers trial (A Randomized Comparison of a Zotarolimus-Eluting Stent With an Everolimus-Eluting Stent for Percutaneous Coronary Intervention) were stratified into "complex" and "simple." RESULTS: Of 2,292 patients, 1,520 (66.3%) were complex and treated with ZES (n = 764) or EES (n = 756). Event rates were higher among complex patients, and results did not differ between ZES and EES, regardless of complexity. At 1 year, target lesion failure was 8.9% in ZES- and 9.7% in EES-treated complex patients (p = 0.66) and 6.8% in ZES- and 5.7% in EES-treated simple patients (p = 0.55). Rates of cardiac death (1.3% vs. 2.2%, p = 0.24), target-vessel myocardial infarction (4.3% vs. 4.4%, p = 0.90), and clinically indicated target lesion revascularization (4.4% vs. 4.0%, p = 0.80) were similar for both stent types among complex patients. Definite or probable stent thrombosis occurred in 20 (1.3%) complex patients with no difference between ZES (1.7%) and EES (0.9%, p = 0.26). Angiographic follow-up showed similar results for ZES and EES in terms of in-stent percentage diameter stenosis (22.2 +/- 15.4% vs. 21.4 +/- 15.8%, p = 0.67) and in-segment binary restenosis (6.6% vs. 8.0%, p = 0.82) in the complex group. CONCLUSIONS: In this all-comers randomized trial, major adverse cardiovascular events were more frequent among complex than simple patients. The newer-generation ZES and EES proved to be safe and effective, regardless of complexity, with similar clinical and angiographic outcomes for both stent types through 1 year. (RESOLUTE-III All Comers Trial: A Randomized Comparison of a Zotarolimus-Eluting Stent With an Everolimus-Eluting Stent for Percutaneous Coronary Intervention; NCT00617084). PMID- 21616283 TI - Volume status and diuretic therapy in systolic heart failure and the detection of early abnormalities in renal and tubular function. AB - OBJECTIVES: This study sought to determine the pharmacodynamic effect of modulation of volume status by withdrawal and reinstitution of diuretic treatment on markers of renal and tubular function. BACKGROUND: Decreased renal perfusion and increased congestion are associated with renal dysfunction in patients with heart failure. METHODS: In this study, 30 patients with chronic systolic heart failure in a presumed euvolemic state and on standard oral furosemide therapy (40 to 80 mg) were examined. At baseline, subjects were withdrawn from their loop diuretics. After 72 h, their furosemide regimen was reinstated, and patients were studied again 3 days later. Serum creatinine, atrial and B-type natriuretic peptide, urinary kidney injury molecule (KIM)-1, urinary N-acetyl-beta-D glucosaminidase (NAG), and serum as well as urinary neutrophil gelatinase associated lipocalin (NGAL) were determined at various time points. RESULTS: Diuretic withdrawal resulted in increases in atrial and B-type natriuretic peptide (both p < 0.05). Serum creatinine was unaffected. Both urinary KIM-1 (p < 0.001) and NAG (p = 0.010) concentrations rose significantly, after diuretic withdrawal, whereas serum and urinary NGAL were not significantly affected. After reinitiation of furosemide, both urinary KIM-1 and NAG concentrations returned to baseline (both p < 0.05), but NGAL values were unaffected. CONCLUSIONS: Subclinical changes in volume status by diuretic withdrawal and reinstitution are associated with increases and decreases of markers of tubular dysfunction in stable heart failure. Diuretic therapy may favorably affect renal and tubular function by decreasing congestion. PMID- 21616284 TI - Diuretics: are our ideas based on knowledge? PMID- 21616285 TI - Flecainide therapy reduces exercise-induced ventricular arrhythmias in patients with catecholaminergic polymorphic ventricular tachycardia. AB - OBJECTIVES: This study evaluated the efficacy and safety of flecainide in addition to conventional drug therapy in patients with catecholaminergic polymorphic ventricular tachycardia (CPVT). BACKGROUND: CPVT is an inherited arrhythmia syndrome caused by gene mutations that destabilize cardiac ryanodine receptor Ca(2+) release channels. Sudden cardiac death is incompletely prevented by conventional drug therapy with beta-blockers with or without Ca(2+) channel blockers. The antiarrhythmic agent flecainide directly targets the molecular defect in CPVT by inhibiting premature Ca(2+) release and triggered beats in vitro. METHODS: We collected data from every consecutive genotype-positive CPVT patient started on flecainide at 8 international centers before December 2009. The primary outcome measure was the reduction of ventricular arrhythmias during exercise testing. RESULTS: Thirty-three patients received flecainide because of exercise-induced ventricular arrhythmias despite conventional (for different reasons, not always optimal) therapy (median age 25 years; range 7 to 68 years; 73% female). Exercise tests comparing flecainide in addition to conventional therapy with conventional therapy alone were available for 29 patients. Twenty two patients (76%) had either partial (n = 8) or complete (n = 14) suppression of exercise-induced ventricular arrhythmias with flecainide (p < 0.001). No patient experienced worsening of exercise-induced ventricular arrhythmias. The median daily flecainide dose in responders was 150 mg (range 100 to 300 mg). During a median follow-up of 20 months (range 12 to 40 months), 1 patient experienced implantable cardioverter-defibrillator shocks for polymorphic ventricular arrhythmias, which were associated with a low serum flecainide level. In 1 patient, flecainide successfully suppressed exercise-induced ventricular arrhythmias for 29 years. CONCLUSIONS: Flecainide reduced exercise-induced ventricular arrhythmias in patients with CPVT not controlled by conventional drug therapy. PMID- 21616286 TI - The V(2) transition ratio: a new electrocardiographic criterion for distinguishing left from right ventricular outflow tract tachycardia origin. AB - OBJECTIVES: We sought to develop electrocardiography (ECG) criteria for distinguishing left ventricular outflow tract (LVOT) from right ventricular outflow tract (RVOT) origin in patients with idiopathic outflow tract ventricular tachycardia (OTVT) and lead V(3) R/S transition. BACKGROUND: Several ECG criteria have been proposed for differentiating left from right OTVT origin; ventricular tachycardias (VTs) with left bundle branch block and V(3) transition remain a challenge. METHODS: We analyzed the surface ECG pattern of patients with OTVT with a precordial transition in lead V(3) who underwent successful catheter ablation. Sinus and VT QRS morphologies were measured in limb and precordial leads with electronic calipers. The V(2) and V(3) transition ratios were calculated by computing the percentage R-wave during VT (R/R+S)(VT) divided by the percentage R-wave in sinus rhythm (R/R+S)(SR). RESULTS: We retrospectively analyzed ECGs from 40 patients (mean age 44 +/- 14 years, 21 female) with outflow tract premature ventricular contractions (PVCs)/VT. Patients with structural heart disease, paced rhythms, and bundle branch block during sinus rhythm were excluded. The V(2) transition ratio was significantly greater for LVOT PVCs compared with RVOT PVCs (1.27 +/- 0.60 vs. 0.23 +/- 0.16; p < 0.001) and was the only independent predictor of LVOT origin. In 21 prospective cases, a V(2) transition ratio >=0.60 predicted an LVOT origin with 91% accuracy. A PVC precordial transition occurring later than the sinus rhythm transition excluded an LVOT origin with 100% accuracy. CONCLUSIONS: The V(2) transition ratio is a novel electrocardiographic measure that reliably distinguishes LVOT from RVOT origin in patients with lead V(3) precordial transition. This measure might be useful for counseling patients and planning an ablation strategy. PMID- 21616287 TI - The utility of cardiac biomarkers, tissue velocity and strain imaging, and cardiac magnetic resonance imaging in predicting early left ventricular dysfunction in patients with human epidermal growth factor receptor II-positive breast cancer treated with adjuvant trastuzumab therapy. AB - OBJECTIVES: The aim of this study was to evaluate whether cardiac biomarkers, tissue velocity (TVI) and strain imaging, and cardiac magnetic resonance imaging can predict early left ventricular (LV) dysfunction in human epidermal growth factor receptor II-positive breast cancer patients treated with trastuzumab in the adjuvant setting. BACKGROUND: Early indexes of LV systolic dysfunction with noninvasive cardiac imaging would be useful for addressing the cardiac safety profile of trastuzumab, potentially avoiding the detrimental effects of heart failure. METHODS: We used cardiac biomarkers, TVI and strain imaging, and cardiac magnetic resonance imaging to detect pre-clinical changes in LV systolic function, before conventional changes in left ventricular ejection fraction (LVEF) in human epidermal growth factor receptor II-positive breast cancer patients treated with trastuzumab in the adjuvant setting. RESULTS: Of 42 patients (mean age 47 +/- 9 years) prospectively followed between 2007 and 2009, 10 (25%) developed trastuzumab-mediated cardiomyopathy (CM). Troponin T, C reactive protein, and brain natriuretic peptide did not change over time. Within 3 months of adjuvant therapy with trastuzumab, there was a significant difference in the lateral S' between the normal cohort and the CM group (9.1 +/- 1.6 cm/s and 6.4 +/- 0.6 cm/s, respectively, p < 0.05). Similarly, the peak global longitudinal and radial strain decreased as early as 3 months in the trastuzumab mediated cardiotoxicity group. As compared with both global longitudinal and radial strain, only S' was able to identify all 10 patients who developed trastuzumab-mediated CM. The LVEF subsequently decreased at 6 months of follow-up in all 10 patients, necessitating discontinuation of the drug. All 10 patients demonstrated delayed enhancement of the lateral wall of the LV within the mid myocardial portion, consistent with trastuzumab-induced CM. CONCLUSIONS: Both TVI and strain imaging were able to detect pre-clinical changes in LV systolic function, before conventional changes in LVEF, in patients receiving trastuzumab in the adjuvant setting. PMID- 21616288 TI - Can modern echocardiographic techniques predict drug-induced cardiotoxicity? PMID- 21616290 TI - Cyanosis in a primum atrial septal defect without pulmonary hypertension. PMID- 21616289 TI - Cardiac-specific overexpression of caveolin-3 attenuates cardiac hypertrophy and increases natriuretic peptide expression and signaling. AB - OBJECTIVES: We hypothesized that cardiac myocyte-specific overexpression of caveolin-3 (Cav-3), a muscle-specific caveolin, would alter natriuretic peptide signaling and attenuate cardiac hypertrophy. BACKGROUND: Natriuretic peptides modulate cardiac hypertrophy and are potential therapeutic options for patients with heart failure. Caveolae, microdomains in the plasma membrane that contain caveolin proteins and natriuretic peptide receptors, have been implicated in cardiac hypertrophy and natriuretic peptide localization. METHODS: We generated transgenic mice with cardiac myocyte-specific overexpression of caveolin-3 (Cav-3 OE) and also used an adenoviral construct to increase Cav-3 in cardiac myocytes. RESULTS: The Cav-3 OE mice subjected to transverse aortic constriction had increased survival, reduced cardiac hypertrophy, and maintenance of cardiac function compared with control mice. In left ventricle at baseline, messenger ribonucleic acid for atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) were increased 7- and 3-fold, respectively, in Cav-3 OE mice compared with control subjects and were accompanied by increased protein expression for ANP and BNP. In addition, ventricles from Cav-3 OE mice had greater cyclic guanosine monophosphate levels, less nuclear factor of activated T cell nuclear translocation, and more nuclear Akt phosphorylation than ventricles from control subjects. Cardiac myocytes incubated with Cav-3 adenovirus showed increased expression of Cav-3, ANP, and Akt phosphorylation. Incubation with methyl-beta-cyclodextrin, which disrupts caveolae, or with wortmannin, a PI3K inhibitor, blocked the increase in ANP expression. CONCLUSIONS: These results imply that cardiac myocyte-specific Cav-3 OE is a novel strategy to enhance natriuretic peptide expression, attenuate hypertrophy, and possibly exploit the therapeutic benefits of natriuretic peptides in cardiac hypertrophy and heart failure. PMID- 21616291 TI - Carotid intima-media thickness as a surrogate endpoint. PMID- 21616292 TI - Carotid intima-media thickness progression and cardiovascular disease risk. PMID- 21616295 TI - Admission hyperlactatemia in intensive care units and mortality. PMID- 21616297 TI - Complexity at the bedside. PMID- 21616298 TI - Proceedings from the Montebello Round Table Discussion. Second annual conference on Complexity and Variability discusses research that brings innovation to the bedside. PMID- 21616429 TI - Radiofrequency ablation of lung metastases close to large vessels during vascular occlusion: preliminary experience. AB - PURPOSE: To report an initial prospective evaluation of the technical feasibility, efficacy, and safety of combining percutaneous temporary balloon occlusion (PBO) of a large pulmonary artery adjacent to a metastatic lung tumor treated with percutaneous radiofrequency (RF) ablation. MATERIALS AND METHODS: In six patients, lung RF ablation with a multitined, expandable electrode with simultaneous PBO via femoral access was attempted with the use of digital angiography and multidetector computed tomography (CT). Follow-up imaging was obtained immediately after treatment, at 1-2 days, and at 2, 6, 9, and 12 months; positron emission tomography/CT was performed at 4 months. RESULTS: Metastases targeted measured 17-37 mm (22 +/- 8) and were in contact with a pulmonary artery 3-5 mm. Temporary occlusion of the pulmonary arterial branch in contact with the tumor was technically possible in five of six patients. Postablation CT scans obtained within 2 days of the procedure showed ablation zones measuring 37-57 mm (47 +/- 8) in their shortest diameter. Three patients developed lung infarction within 1 month after RF ablation, and two had to be readmitted. At 3 months after the procedure, four patients had persistent occlusion of the balloon-occluded vessel. No uptake was demonstrated 4 months after ablation; at 12 months, all tumors showed complete ablation on CT. CONCLUSIONS: RF ablation of lung tumors with PBO is a feasible technique, but it induces atelectasia and long-lasting vascular occlusion responsible for a high rate of readmission. The results of this small study warrant careful further exploration of the benefits of the technique, compared with RF ablation without PBO or other methods of ablative therapy. PMID- 21616430 TI - A comparison of US-guided percutaneous radiofrequency ablation of medium-sized hepatocellular carcinoma with a cluster electrode or a single electrode with a multiple overlapping ablation technique. AB - PURPOSE: To compare ablation zone, local therapeutic efficacy, and complications of ultrasound (US)-guided percutaneous radiofrequency (RF) ablation of medium sized hepatocellular carcinomas (HCCs) with a cluster electrode versus a single electrode with multiple overlapping ablations. MATERIALS AND METHODS: From February 2005 to January 2009, a total of 79 consecutive patients (57 men, 22 women; mean age, 58.6 y) with 79 HCCs (range, 2.5-4.0 cm) underwent percutaneous RF ablation with a cluster electrode (n = 37) or a single electrode with multiple overlapping ablations (n = 42). These methods were compared in terms of ablation zone size on immediate follow-up computed tomography and the rates of technique effectiveness and cumulative local tumor progression on further follow-up (range, 12.0-46.2 mo; median, 24.3 mo). RESULTS: Baseline characteristics did not differ between groups. The ablation zone in the cluster group was significantly smaller in the longitudinal dimension (43.7 mm +/- 4.8 vs 46.5 mm +/- 5.9; P = .03), but greater in the short axial dimension (30.7 mm +/- 3.9 vs 27.3 mm +/- 5.5; P = .03), compared with that in the overlapping ablation group. Technique effectiveness rates in the cluster and overlap groups were 100% and 92.9% (39 of 42), respectively (P = .24). The cumulative local tumor progression rate was significantly lower in the cluster group than in the overlapping group (1- and 3- year follow-up: 8.1 and 18.8% vs 23.8 and 42.2%, P = .04). Complications were more frequent in the cluster group than in the overlapping group (27.0% vs 7.1%; P = .03). CONCLUSIONS: In US-guided percutaneous RF ablation of medium-sized HCCs, the cluster electrode showed better local therapeutic efficacy than the single electrode with multiple overlapping ablations, probably because of the favorable shape of the ablation zone; however, complications occurred more frequently. PMID- 21616431 TI - Safety and efficacy of radiofrequency ablation for hepatocellular adenoma. AB - PURPOSE: To investigate the safety and efficacy of radiofrequency (RF) ablation for the treatment of hepatocellular adenoma (HCA). MATERIALS AND METHODS: From 2000 to 2009, 170 patients with HCA were referred to a single tertiary hepatobiliary center. Medical records of 18 patients treated with RF ablation were retrospectively analyzed. RESULTS: All patients were female, and the majority had a history of hormonal contraceptive use. Ten patients (56%) had multiple HCAs, with a median number of two lesions (range, one to 12) per patient. Median size of HCA at the time of RF ablation was 3.0 cm (range, 0.8-7.3 cm). A total of 45 HCAs were ablated in 32 sessions (open, n = 4; percutaneous, n = 28). RF ablation was complete after the first session in 26 HCAs (57.8%), and the majority of patients underwent multiple RF ablation sessions to fully ablate all HCAs. Major complications developed in two patients. CONCLUSIONS: RF ablation can be used effectively in the treatment of HCA. However, multiple sessions are often required, and signs of residual adenoma might persist in some patients despite repetitive treatment. RF ablation might be especially beneficial in cases not amenable to surgery or in patients who would require major hepatic resection otherwise. PMID- 21616432 TI - Bland embolization in the treatment of hepatic adenomas: preliminary experience. AB - The study presents preliminary, retrospective experience with bland embolization for hepatic adenomas (HAs) with the use of tris-acryl gelatin microspheres. Eight patients underwent bland embolization for biopsy-proven HAs. A biopsy specimen was taken from only one lesion when multiple lesions were present. Seventeen embolizations were performed for abdominal pain, active bleeding, or prophylaxis against bleeding. Five patients underwent multiple procedures. Technical success rate was 100%. Median follow-up was 24 months (range, 10-40 mo). No growth was observed at follow-up in any treated lesion, and many lesions (13 of 16) regressed. Bland embolization may be safely used for treatment of abdominal pain and bleeding from HAs. PMID- 21616433 TI - Host defense pathways: role of redundancy and compensation in infectious disease phenotypes. AB - Innate host defense pathways consist of microbial sensors, their signaling pathways, and the antimicrobial effector mechanisms. Several classes of host defense pathways are currently known, each comprising several pattern-recognition receptors that detect different types of pathogens. These pathways interact with one another in a variety of ways that can be categorized into cooperation, complementation, and compensation. Understanding the principles of these interactions is important for better understanding of host defense mechanisms, as well as for correct interpretation of immunodeficient phenotypes. PMID- 21616434 TI - Toll-like receptors and their crosstalk with other innate receptors in infection and immunity. AB - Toll-like receptors (TLRs) are germline-encoded pattern recognition receptors (PRRs) that play a central role in host cell recognition and responses to microbial pathogens. TLR-mediated recognition of components derived from a wide range of pathogens and their role in the subsequent initiation of innate immune responses is widely accepted; however, the recent discovery of non-TLR PRRs, such as C-type lectin receptors, NOD-like receptors, and RIG-I-like receptors, suggests that many aspects of innate immunity are more sophisticated and complex. In this review, we will focus on the role played by TLRs in mounting protective immune responses against infection and their crosstalk with other PRRs with respect to pathogen recognition. PMID- 21616435 TI - Myeloid C-type lectin receptors in pathogen recognition and host defense. AB - C-type lectin receptors (CLRs) comprise a heterogeneous group of transmembrane proteins. Many of them are expressed in myeloid cells and signal in response to pathogen-derived or self ligands to initiate or regulate cell activation. Here, we review the properties of myeloid CLRs, highlighting how their signaling function is coordinated with that of other innate receptor families to control immunity to infection. PMID- 21616436 TI - Regulation of the antimicrobial response by NLR proteins. AB - Nucleotide-binding, oligomerization domain (NOD)-like receptor (NLR) proteins are a family of innate immune receptors that play a pivotal role in microbial sensing, leading to the initiation of antimicrobial immune responses. Dysregulation of the function of multiple NLR family members has been linked, both in mice and humans, to a propensity for infection and autoinflammatory disease. Despite our increased understanding of NLR function and interactions, many aspects related to mechanisms of sensing, downstream signaling, and in vivo functions remain elusive. In this review, we focus on key members of the NLR family, describing their activation by diverse microbes, downstream effector functions, and interactions with each other and with other innate sensor protein families. Also discussed is the role of microbial sensing by NLR receptors leading to activation of the adaptive immune arm that collaborates in the antimicrobial defense. PMID- 21616438 TI - Gut Nod2 calls the bone marrow for monocyte reinforcements. AB - Nod2 is an intracellular sensor linked to Crohn's disease, an inflammatory malady of the intestinal tract. In this issue of Immunity, Kim et al. (2011) demonstrate that Nod2 is responsible for regulating monocyte-attracting chemokines to the inflamed gut. PMID- 21616439 TI - A new twist on the PYRIN Mediterranean coast. AB - Familial Mediterranean fever is caused by mutations of the PYRIN protein. Chae et al. (2011) provide evidence for a ASC protein-dependent pathway of caspase-1 activation in which gain-of-function PYRIN mutations lead to IL-1beta cytokine overproduction and inflammatory disease. PMID- 21616440 TI - A narrow circle of mutual friends. AB - Commensal microbiota confers a goldilocks state of alertness to pathogens, yet restrains deleterious inflammation. In this issue of Immunity, Geuking et al. (2011) demonstrate that a minimal bacterial community of the Schaedler flora establishes a balance between pro- and anti-inflammatory T cells in the gut. PMID- 21616441 TI - Chronic infections capture little attention of the masses. AB - In this issue of Immunity, Egen et al. (2011) provide compelling evidence that only a minute fraction of mycobacteria-specific T cells present in a granuloma are actively fulfilling effector functions, an observation that may in fact be a general feature of chronic infections. PMID- 21616442 TI - [Controversies, challenges and debate: how to build the future of urology?]. PMID- 21616437 TI - Immune signaling by RIG-I-like receptors. AB - The RIG-I-like receptors (RLRs) RIG-I, MDA5, and LGP2 play a major role in pathogen sensing of RNA virus infection to initiate and modulate antiviral immunity. The RLRs detect viral RNA ligands or processed self RNA in the cytoplasm to trigger innate immunity and inflammation and to impart gene expression that serves to control infection. Importantly, RLRs cooperate in signaling crosstalk networks with Toll-like receptors and other factors to impart innate immunity and to modulate the adaptive immune response. RLR regulation occurs at a variety of levels ranging from autoregulation to ligand and cofactor interactions and posttranslational modifications. Abberant RLR signaling or dysregulation of RLR expression is now implicated in the development of autoimmune diseases. Understanding the processes of RLR signaling and response will provide insights to guide RLR-targeted therapeutics for antiviral and immune modifying applications. PMID- 21616443 TI - [Ongoing debates on localized prostate cancer: epidemiology, screening and stratification]. AB - Diagnosis and treatment of localized prostate cancer is debated within the urological community. While the incidence of the disease has increased significantly due to the screening, recent data from the literature showed that indiscriminate support led to a high risk of over-treatment and unnecessary morbidity. The development of new strategies for detection and diagnosis, based on the progress of biology (pro-PSA, urinary PCA3, genetic polymorphism) and imaging (diffusion MRI), helps to better characterize localized prostate cancers and to better adapt their support. At the 2010 AFU Congress, many studies aimed to better target patients at risk of cancer within the population and to better identify cancers with a high potential of evolution among the diagnosed cancers. This requires a better knowledge of risk factors (age, family history, black race, exposition to steroid hormones), the definition of finer criteria for targeted screening and a more accurate staging to make an appropriate therapeutic response. PMID- 21616444 TI - [News and perspective in management of high-risk prostate cancer]. AB - Advanced prostate cancers include situations involving the patient's survival in the medium or short run. They are responsible for the second leading cause of death by cancer in France. However, their management is poorly codified and confronts clinicians and researchers to therapeutic problems. Even if the main options remain surgery radiotherapy and hormone therapy, alone or in combination, their places tends to be clarified. The association radio-hormonotherapy is the gold standard, but surgery first has interesting results with the possibility of multi-modal salvage treatment if necessary. Similarly, improved diagnostic tools allow a more personal management of these patients and the search for predictive factors (early PSA assay, time to reach PSA nadir) contribute to provide better monitoring. The mortality of prostate cancer decreases by 2.5 % per year due to earlier detection of cancer and advances in treatment of advanced forms. Technical advances in surgery and radiotherapy, associated with new ways of androgeno-suppression should continue this trend. PMID- 21616445 TI - [Surgical innovations and therapeutic strategies in the management of renal cell carcinomas]. AB - In recent years, the management of renal cell carcinomas has changed significantly and is now based on two concepts: preservation of renal function in localized forms and control of angiogenesis in metastatic forms. The main ways of research in the diagnostic strategy is to develop new tools to identify more precisely the lesions discovered by imaging, thanks to biological or molecular markers as micro-RNA, and to predict renal function after surgery. Surgical indications for partial nephrectomy are now only limited by technical feasibility. In this regard, tolls like the RENAL nephrometric score help to evaluate the risk of total nephrectomy or the duration of clamping. Furthermore, the important development of robotically assisted surgery provides very promising results. In advanced forms of renal cell carcinomas, recent advances have led to a considerable prolongation of recurrence-free survival, but overall survival remains the same with 40 % of specific mortality. New targeted therapies have paved the way for a new approach and studies expected in the coming years could change the therapeutic strategies and lead to a new staging of patients. PMID- 21616446 TI - [How to select the best candidates for prostate biopsies? The role of traditional tools and contribution of new biomarkers in prostate cancer]. AB - In 2010, early detection of prostate cancer continues to rely on digital rectal examination and serum total PSA. However, selecting patients for prostate biopsy requires taking into account the prostate volume and the evolution of PSA over time. PSA derivatives such as PSA density, PSA velocity and the ratio free PSA / total PSA are useful supplements. However, the choice of the threshold value is not well defined and depends on the relative sensitivity and specificity desired. The real innovations come from basic research that has found potential markers of aggressiveness of prostate cancer and molecular biology tools used routinely as the PCA-3 score and the pro-PSA. The role of these new markers for diagnosis and prognosis of prostate cancer remains unclear. PMID- 21616447 TI - [Evolution of modern imaging and development of targeted prostatic biopsies for the diagnosis of prostate cancer]. AB - Prostatic biopsies are the standard procedure to diagnose of prostate cancer and provide historical prognostic criteria. However the prostatic biopsies are relatively blind and the echography is more useful to identify the organ than to target the tumour. Advances in imaging in recent years with the development of 3D can allow new practices. Several tools exist and are being evaluated to allow recording the location of prostate biopsies under echography (mapping) and to guide biopsy, possibly by fusing MRI images. Other tools are being developed for the implementation of robotics biopsies under ultrasound or MRI. PMID- 21616448 TI - [Innovations in hormonal treatment for locally advanced and/or metastatic prostate cancer]. AB - Discovered over 40 years ago, hormonal therapy remains the cornerstone therapy of advanced prostate cancer and continues to evolve. Suppression of serum testosterone remains the mainstay of systemic treatment of prostate cancer. Antagonists of LH-RH are now available and can prevent the castration delay of agonists. They seem to have a clinical benefit in terms of PSA control. During the phase of resistance to extracellular castration, the androgen receptor is the dominant element. Intracellular steroidogenesis can be blocked by Abiraterone. Amplification and mutation of the androgen receptor may be controlled by antiandrogens of second generation. Thus, these new molecules, already or soon available, will renew the strategy of prostate cancer treatment. PMID- 21616449 TI - One health for 7 billion. PMID- 21616450 TI - Is screening immigrants for latent tuberculosis cost-effective? PMID- 21616451 TI - How common was 2009 pandemic influenza A H1N1? PMID- 21616452 TI - Prevalence of mycoplasma encephalitis. PMID- 21616453 TI - Do children receiving HAART need to be revaccinated? PMID- 21616454 TI - Polymyxin B for endotoxin removal in sepsis. PMID- 21616455 TI - Vaccination against meningococcal meningitis outside the meningitis belt. PMID- 21616456 TI - Leprosy now: epidemiology, progress, challenges, and research gaps. AB - Leprosy continues to be a challenge to health worldwide, with about 250,000 new cases being detected every year. Despite widespread implementation of effective multidrug therapy, leprosy has not been eliminated. A third of newly diagnosed patients have nerve damage and might develop disabilities, although the proportion varies according to several factors, including level of self-care. Women who develop leprosy continue to be especially disadvantaged, with rates of late diagnosis and disability remaining high in this subgroup. Leprosy was not a specified disease in the Millennium Development Goals, but improvements in the other areas they cover, such as education and levels of poverty, will help leprosy patients and services. We review data and make recommendations for research on diagnosis, treatment, and prevention, such as further use of molecular analysis of the Mycobacterium leprae genome, implementation of BCG vaccination, and administration of chemoprophylaxis to household contacts. We also suggest development of tools for early diagnosis and detection of infection and nerve damage, and formulation of strategies to manage the chronic complications of leprosy, such as immune-mediated reactions and neuropathy. PMID- 21616457 TI - Use of benchmarking and public reporting for infection control in four high income countries. AB - Benchmarking of surveillance data for health-care-associated infection (HCAI) has been used for more than three decades to inform prevention strategies and improve patients' safety. In recent years, public reporting of HCAI indicators has been mandated in several countries because of an increasing demand for transparency, although many methodological issues surrounding benchmarking remain unresolved and are highly debated. In this Review, we describe developments in benchmarking and public reporting of HCAI indicators in England, France, Germany, and the USA. Although benchmarking networks in these countries are derived from a common model and use similar methods, approaches to public reporting have been more diverse. The USA and England have predominantly focused on reporting of infection rates, whereas France has put emphasis on process and structure indicators. In Germany, HCAI indicators of individual institutions are treated confidentially and are not disseminated publicly. Although evidence for a direct effect of public reporting of indicators alone on incidence of HCAIs is weak at present, it has been associated with substantial organisational change. An opportunity now exists to learn from the different strategies that have been adopted. PMID- 21616458 TI - New approaches to the assessment of vaccine herd protection in clinical trials. AB - Criteria for the introduction of new vaccines into routine public health practice are becoming increasingly stringent. For vaccines that are expensive and those that provide moderate protection, the ability to confer herd protection could be crucial to policy deliberations about vaccine introduction. Traditionally, herd protection has been assessed after a vaccine is introduced, delaying the availability of data on herd effects to inform decisions about vaccine introduction. New methodological developments now provide the possibility to assess herd protection before the introduction of a vaccine into public health programmes. One approach is a cluster-randomised trial, which allows assessment of herd protection in a way that minimises biases. Analysis of individually randomised trials by appropriately selected clusters created post hoc can also provide measurements of herd protection. Here we discuss the use of these designs, which can generate an improved evidence base at an early stage for making decisions about the introduction of new vaccines. PMID- 21616459 TI - Umbilicated erythematous papules in an immunocompromised patient. PMID- 21616460 TI - Screen the information on screenings. PMID- 21616461 TI - Objective diagnostic and interventional vision test protocol for the mild traumatic brain injury population. PMID- 21616462 TI - Paraoptometry has a history of vision. PMID- 21616463 TI - Bone remodeling. PMID- 21616464 TI - Multiscale mechanobiology of de novo bone generation, remodeling and adaptation of autograft in a common ovine femur model. AB - The link between mechanics and biology in the generation and the adaptation of bone has been studied for more than a century in the context of skeletal development and fracture healing. However, the interplay between mechanics and biology in de novo generation of bone in postnatal defects as well as healing of morcellized bone graft or massive cortical bone autografts is less well understood. To address this, here we integrate insights from our previously published studies describing the mechanobiology on both de novo bone generation and graft healing in a common ovine femoral defect model. Studying these effects in a common experimental model provides a unique opportunity to elucidate factors conducive to harnessing the regenerative power of the periosteum, and ultimately, to provide mechanistic insights into the multiscale mechanobiology of bone generation, remodeling and adaptation. Taken together, the studies indicate that, as long as adequate, directional transport of cells and molecules can be insured (e.g. with periosteum in situ or a delivery device), biological factors intrinsic to the periosteum suffice to bridge critical sized bone defects, even in the absence of a patent blood supply. Furthermore, mechanical stimuli are crucial for the success of periosteal bone generation and bone graft healing. Interestingly, areas of highest periosteal strain around defects correlate with greatest amounts albeit not greatest mineralization of newly generated bone. This may indicate a role for convection enhanced transport of cells and molecules in modulation of tissue generation by pluripotent cells that ingress into the defect center, away from the periosteum and toward the surface of the intramedullary nail that fills the medullary cavity. These insights bring us much closer to understanding the mechanobiological environment and stimuli that stimulate the proliferation and differentiation of periosteum-derived progenitor cells and ultimately drive the generation of new bone tissue. Furthermore, these insights provide a foundation to create virtual predictive computational models of bone mechanophysiology, to develop cell seeding protocols for scale up and manufacture of engineered tissues, to optimize surgical procedures, and to develop post-surgical therapies with the ultimate goal of achieving the best possible healing outcomes for treatment and/or reconstruction of postnatal bone defects. PMID- 21616465 TI - Hierarchies of damage induced loss of mechanical properties in calcified bone after in vivo fatigue loading of rat ulnae. AB - During fatigue loading of whole bone, damage to bone tissue accumulates, coalesces and leads to fractures. Whether damage affects tissue material properties similarly at the nanoscale (less than 1 MUm), microscale (less than 1 mm), and whole bone scale has not been fully evaluated. Therefore, in this study, we examine scale-dependent loss of calcified tissue material properties in rat ulnae, after fatigue loading of rat forearms using the forearm compression model. In vivo fatigue loading was conducted on the right forearms until a displacement end-point was reached. The non-fatigued left forearms served as contralateral controls. Subsequently, three-point bending tests to failure on excised ulnae demonstrated a 41% and 49% reduction in the stiffness and ultimate strength as compared to contralateral control ulnae, respectively. Depth-sensing microindentation demonstrated an average decrease in material properties, such as elastic modulus and hardness, of 28% and 29% respectively. Nanoindentation measured elastic modulus and hardness were reduced by 26% and 29% in damaged bone relative to contralateral controls, respectively. The increased loss of whole bone material properties compared to tissue material properties measured using indentation is mainly attributed to the presence of a macrocrack located in the medial compressive region at the site of peak strains. The similar magnitude of changes in material properties by microindentation and nanoindentation is attributed to damage that may originate at an even smaller scale, as inferred from 10% differences in connectivity of osteocyte canaliculi in damaged bone. PMID- 21616466 TI - Numerical model of bone remodeling sensitive to loading frequency through a poroelastic behavior and internal fluid movements. AB - In this article, a phenomenological numerical model of bone remodeling is proposed. This model is based on the poroelasticity theory in order to take into account the effects of fluid movements in bone adaptation. Moreover, the proposed remodeling law is based on the classical 'Stanford' law, enriched in order to take into account the loading frequency, through fluid movements. This coupling is materialized by a quadratic function of Darcy velocity. The numerical model is carried out, using a finite element method, and calibrated using experimental results at macroscopic level, from the literature. First results concern cyclic loadings on a mouse ulna, at different frequencies between 1 Hz and 30 Hz, for a force amplitude of 1.5 N and 2 N. Experimental results exhibit a sensitivity to the loading frequency, with privileged frequency for bone remodeling between 5 Hz and 10 Hz, for the force amplitude of 2 N. For the force amplitude of 1.5 N, no privileged frequencies for bone remodeling are highlighted. This tendency is reproduced by the proposed numerical computations. The model is identified on a single case (one frequency and one force amplitude) and validated on the other ones. The second experimental validation deals with a different loading regime, an internal fluid pressure at 20 Hz on a turkey ulna. The same framework is applied, and the numerical and experimental data are still matching in terms of gain in bone mass density. PMID- 21616467 TI - Efficient numerical analysis of bone remodelling. AB - This paper presents a formulation for the three-dimensional numerical simulation of mechanically regulated bone adaptation. Attention is focussed on a phenomenologically-based approach to bone remodelling that can be used as a computationally efficient tool to provide insight into the overall response of bone to mechanical loading. A discretisation approach is developed based on a hybrid finite element formulation where displacement, stress and density fields are approximated independently. The paper also discusses a solution algorithm tailored for shared memory multi-core computers. The performance of the model is demonstrated by two numerical examples. PMID- 21616468 TI - Apparent damage accumulation in cancellous bone using neural networks. AB - In this paper, a neural network model is developed to simulate the accumulation of apparent fatigue damage of 3D trabecular bone architecture at a given bone site during cyclic loading. The method is based on five steps: (i) performing suitable numerical experiments to simulate fatigue accumulation of a 3D micro-CT trabecular bone samples taken from proximal femur for different combinations of loading conditions; (ii) averaging the sample outputs in terms of apparent damage at whole specimen level based on local tissue damage; (iii) preparation of a proper set of corresponding input-output data to train the network to identify apparent damage evolution; (iv) training the neural network based on the results of step (iii); (v) application of the neural network as a tool to estimate rapidly the apparent damage evolution at a given bone site. The proposed NN model can be incorporated into finite element codes to perform fatigue damage simulation at continuum level including some morphological factors and some bone material properties. The proposed neural network based multiscale approach is the first model, to the author's knowledge, that incorporates both finite element analysis and neural network computation to rapidly simulate multilevel fatigue of bone. This is beneficial to develop enhanced finite element models to investigate the role of damage accumulation on bone damage repair during remodelling. PMID- 21616469 TI - Trabecular bone remodelling simulated by a stochastic exchange of discrete bone packets from the surface. AB - Human bone is constantly renewed through life via the process of bone remodelling, in which individual packets of bone are removed by osteoclasts and replaced by osteoblasts. Remodelling is mechanically controlled, where osteocytes embedded within the bone matrix are thought to act as mechanical sensors. In this computational work, a stochastic model for bone remodelling is used in which the renewal of bone material occurs by exchange of discrete bone packets. We tested different hypotheses of how the mechanical stimulus for bone remodelling is integrated by osteocytes and sent to actor cells on the bone's surface. A collective (summed) signal from multiple osteocytes as opposed to an individual (maximal) signal from a single osteocyte was found to lead to lower inner porosity and surface roughness of the simulated bone structure. This observation can be interpreted in that collective osteocyte signalling provides an effective surface tension to the remodelling process. Furthermore, the material heterogeneity due to remodelling was studied on a network of trabeculae. As the model is discrete, the age of individual bone packets can be monitored with time. The simulation results were compared with experimental data coming from quantitative back scattered electron imaging by transforming the information about the age of the bone packet into a mineral content. Discrepancies with experiments indicate that osteoclasts preferentially resorb low mineralized, i.e. young, bone at the bone's surface. PMID- 21616470 TI - Multiscale FE method for analysis of bone micro-structures. AB - Bones are composed of hierarchical bio-composite materials characterized by complex multiscale structural geometry and behavior. The architecture and the mechanical properties of bone tissue differ at each level of hierarchy. Thus, a multiscale approach for mechanical analysis of bone is imperative. This paper proposes a new approach for 3D multiscale finite element analysis of trabecular bone that can offer physicians a "digital magnifying glass" to facilitate continuous transition between macro- and micro-scales. The approach imitates the human ability to perceive details. That is, zooming-out from an object causes fewer details to be visible. As a result, the material appears to be smoother and more homogeneous. Zooming-in, in contrast, reveals additional details and material heterogeneity. Realization of the proposed approach requires synergy between a hierarchical geometric model for representing intermediate scales and a mechanical model for local material properties of bone tissue for each scale. The geometric model facilitates seamless and continuous bi-directional transition between macro- and micro-scales, while the mechanical model preserves the effective material properties. A 2D model of a simplified trabecular structure was implemented and analyzed in order to assess the feasibility of the proposed multiscale approach. The successful results of this model led to extending the method into 3D and analyzing real trabecular structures. PMID- 21616471 TI - Effects of loading frequency on the functional adaptation of trabeculae predicted by bone remodeling simulation. AB - The process of bone remodeling is regulated by metabolic activities of many bone cells. While osteoclasts and osteoblasts are responsible for bone resorption and formation, respectively, activities of these cells are believed to be controlled by a mechanosensory system of osteocytes embedded in the extracellular bone matrix. Several experimental and theoretical studies have suggested that the strain-derived interstitial fluid flow in lacuno-canalicular porosity serves as the prime mover for bone remodeling. Previously, we constructed a mathematical model for trabecular bone remodeling that interconnects the microscopic cellular activities with the macroscopic morphological changes in trabeculae through the mechanical hierarchy. This model assumes that fluid-induced shear stress acting on osteocyte processes is a driving force for bone remodeling. The validity of this model has been demonstrated with a remodeling simulation using a two dimensional trabecular model. In this study, to investigate the effects of loading frequency, which is thought to be a significant mechanical factor in bone remodeling, we simulated morphological changes of a three-dimensional single trabecula under cyclic uniaxial loading with various frequencies. The results of the simulation show the trabecula reoriented to the loading direction with the progress of bone remodeling. Furthermore, as the imposed loading frequency increased, the diameter of the trabecula in the equilibrium state was enlarged by remodeling. These results indicate that our simulation model can successfully evaluate the relationship between loading frequency and trabecular bone remodeling. PMID- 21616472 TI - What is the importance of multiphysical phenomena in bone remodelling signals expression? A multiscale perspective. AB - Cortical bone, constituting the outer shell of long bones, is continuously renewed by bone cells in response to daily stimuli. This process, known as bone remodelling, is essential for proper bone functioning in both physiological and pathological conditions. Classical bone remodelling models do not, or only implicitly do, take into account physico-chemical phenomena, focussing on the mechanosensitivity property of the tissue. The aim of this paper is to carry out an investigation of the multiphysical phenomena occuring in bone life. Using a recent multiscale model combining piezoelectricity and electrokinetics to poromechanics, the usual viewpoint of bone remodelling models is questioned and new research avenues are proposed. PMID- 21616473 TI - [Past, present, and future of propranolol for hemangiomas of infancy]. PMID- 21616474 TI - Electrostatic and steric interactions in oil-in-water emulsion films from Pluronic surfactants. AB - Stabilization of oil-in-water emulsion films from PEO-PPO-PEO triblock copolymers is described in terms of interaction surface forces. Results on emulsion films from four Pluronic surfactants, namely F108, F68, P104 and P65 obtained with the Thin Film Pressure Balance Technique are summarized. It is found that film stabilization is due to DLVO (electrostatic) and non-DLVO (steric in origin) repulsive forces. The charging of the oil/water film interfaces is related to preferential adsorption of OH(-) ions. This is confirmed by pH-dependent measurements of the equivalent film thickness (h(w)) at both constant capillary pressure and ionic strength. With reducing pH in the acidic region, a critical value (pH(cr,st)) corresponding to an isoelectric state of the oil/water film surfaces is found where the electrostatic interaction in the films is eliminated. At pH<=pH(cr,st), the emulsion films are stabilized only by steric forces due to interaction between the polymer adsorption layers. Disjoining pressure (Pi) isotherms measured for emulsion films from all the four Pluronic surfactants used at pH50 Gy), and 6 received radiation alone. Two were aborted owing to fibrosis. Stations 4L, 4R, and 7 were accessed in 84% of patients, with confirmed nodal tissue in 88%. There was 1 azygos vein injury that required urgent thoracotomy and 2 recurrent nerve injuries. Resection ensued in 63 patients: 53 with negative mediastinoscopy, 8 with microscopic nodal metastases, and the 2 aborted cases. In patients with negative mediastinoscopy, 5 had N2 disease at thoracotomy: 3 in stations 4 and 7 and 2 in nodal stations inaccessible by mediastinoscopy. The sensitivity was 71%, specificity was 100%, and negative predictive value was 91%. CONCLUSIONS: In experienced hands, mediastinoscopy in a complex mediastinum is safe. Preclusive mediastinal fibrosis is rare. Expected lymph node stations can be accessed, and the results strongly correlate with postresection pathology. Mediastinoscopy is valuable to evaluate the nodal response to therapy in the setting of combined modality therapy. PMID- 21616476 TI - Factors of collaborative working: a framework for a collaboration model. AB - The ability of organisations to support collaborative working environments is of increasing importance as they move towards more distributed ways of working. Despite the attention collaboration has received from a number of disparate fields, there is a lack of a unified understanding of the component factors of collaboration. As part of our work on a European Integrated Project, CoSpaces, collaboration and collaborative working and the factors which define it were examined through the literature and new empirical work with a number of partner user companies in the aerospace, automotive and construction sectors. This was to support development of a descriptive human factors model of collaboration - the CoSpaces Collaborative Working Model (CCWM). We identified seven main categories of factors involved in collaboration: Context, Support, Tasks, Interaction Processes, Teams, Individuals, and Overarching Factors, and summarised these in a framework which forms a basis for the model. We discuss supporting evidence for the factors which emerged from our fieldwork with user partners, and use of the model in activities such as collaboration readiness profiling. PMID- 21616477 TI - Urease activity in dental plaque and saliva of children during a three-year study period and its relationship with other caries risk factors. AB - Bacterial urease activity in dental plaque and in saliva generates ammonia, which can increase the plaque pH and can protect acid-sensitive oral bacteria. Recent cross-sectional studies suggest that reduced ability to generate ammonia from urea in dental plaque can be an important caries risk factor. In spite of this proposed important clinical role, there is currently no information available regarding important clinical aspects of oral ureolysis in children. OBJECTIVE: The objective of this study was to evaluate the distribution and pattern of urease activity in the dental plaque and in the saliva of children during a three year period, and to examine the relationship of urease with some important caries risk factors. METHODS: A longitudinal study was conducted with repeated measures over a three-year period on a panel of 80 children, aged 3-6 years at recruitment. The dynamics of change in urease activity were described and associated with clinical, biological, and behavioural caries risk factors. RESULTS: Urease activity in plaque showed a trend to remain stable during the study period and was negatively associated with sugar consumption (P<0.05). Urease activity in unstimulated saliva increased with age, and it was positively associated with the levels of mutans streptococci in saliva and with the educational level of the parents (P<0.05). CONCLUSIONS: The results of this study reveal interesting and complex interactions between oral urease activity and some important caries risk factors. Urease activity in saliva could be an indicator of mutans infection in children. PMID- 21616478 TI - Cyclin-dependent kinase 5 activator p25 is generated during memory formation and is reduced at an early stage in Alzheimer's disease. AB - BACKGROUND: The cyclin-dependent kinase 5 activator p35 can be cleaved into p25. Formation of p25 has been suggested to contribute to neurodegeneration in Alzheimer's disease (AD). However, overexpression of low levels of p25 in mice enhances memory formation. Therefore, it has been suggested that p25 formation might be an event early in AD to compensate for impairments in synaptic plasticity. Ongoing p25 formation has been hypothesized to contribute to neurodegeneration at the later stages of AD. METHODS: Here, we tested the early compensation hypothesis by analyzing the levels of p25 and its precursor p35 in AD postmortem samples from different brain regions at different stages of tau pathology, using quantitative Western blots. Furthermore, we studied p35 and p25 during spatial memory formation. By employing quantitative mass spectrometry, we identified proteins downstream of p25, which were then studied in AD samples. RESULTS: We found that p25 is generated during spatial memory formation. Furthermore, we demonstrate that overexpression of p25 in the physiological range increases the expression of two proteins implicated in spine formation, septin 7 and optic atrophy 1. We show that the expression of p35 and p25 is reduced as an early event in AD. Moreover, expression of the p25-regulated protein optic atrophy 1 was reduced in a time course similar to p25 expression. CONCLUSIONS: Our findings suggest that p25 generation is a mechanism underlying hippocampal memory formation that is impaired in the early stages of AD. Our findings argue against the previously raised early compensation hypothesis and they propose that p25-mediated neurotoxicity does not occur in AD. PMID- 21616479 TI - Polyunsaturated fatty acids and blood circulation in the forebrain during a mental arithmetic task. AB - The effects of polyunsaturated fatty acids on human cerebral blood oxygenation have yet to be extensively investigated. In this study, healthy participants (14 men, 40 women) aged between 20 and 49 years were recruited. All female participants entered the trial at the start of their menstrual cycle. Blood was sampled before measuring cerebral blood oxygenation in the prefrontal cortex (PFC) and prior to administering two kinds of questionnaires, the Profile of Mood States (POMS) and a questionnaire regarding participants' arousal level. Blood oxygenation in the PFC was continuously monitored immediately before and during the Uchida-Kraepelin Performance (UKP) test as a mental arithmetic task. Changes in the tissue oxygenation index (the ratio of oxyhemoglobin to oxyhemoglobin+deoxyhemoglobin; TOI, a simplified index for cerebral blood circulation) were measured by near-infrared spectroscopy. Multiple regression analysis was performed with sex, age, smoking and drinking as confounding factors. Eicosapentaenoic acid (EPA) was positively associated with TOI, which was positively associated with arousal level and inversely associated with negative mood (POMS). EPA and docosahexaenoic acid were inversely associated with depression-dejection (POMS) and positively associated with arousal level and overall performance in the UKP test. We suggest that EPA might increase the oxygenation level in the PFC, in turn improving various psychological parameters and performance. PMID- 21616480 TI - A kinetic study on the reductive opening of the diphenylmethylene acetal in methyl 2,3-O-diphenylmethylene-alpha-L-rhamnopyranoside. AB - Reductive opening of the diphenylmethyl acetal in methyl 2,3-O-diphenylmethylene alpha-l-rhamnopyranoside has been investigated by kinetic studies, and the results have been compared to those recently obtained by quantum chemical calculations. In contrast to the previous theoretical calculations which related only to the presumably rate limiting step of the reductive opening, the reaction system LiAlH(4), AlCl(3), and the title compound consists of at least four simultaneous reactions. Nevertheless, reasonable agreement can be found between the activation Gibbs free energy obtained from kinetic measurements and the theoretically calculated ones in spite of the experimental errors and the approximate nature of theoretical calculations. PMID- 21616481 TI - [Sigmoid achalasia]. PMID- 21616482 TI - [Redundant coloplasty as a cause of a persistent cervical fistula]. PMID- 21616483 TI - "I had so much it didn't seem fair": Eight-year-olds reject two forms of inequity. AB - Research using economic games has demonstrated that adults are willing to sacrifice rewards in order to prevent inequity both when they receive less than a social partner (disadvantageous inequity) and when they receive more (advantageous inequity). We investigated the development of both forms of inequity aversion in 4- to 8-year-olds using a novel economic game in which children could accept or reject unequal allocations of candy with an unfamiliar peer. The results showed that 4- to 7-year-olds rejected disadvantageous offers, but accepted advantageous offers. By contrast, 8-year-olds rejected both forms of inequity. These results suggest that two distinct mechanisms underlie the development of the two forms of inequity aversion. PMID- 21616484 TI - Tract based spatial statistical analysis and voxel based morphometry of diffusion indices in temporal lobe epilepsy. AB - White matter (WM) microstructure can be evaluated by diffusion tensor imaging (DTI). Tract-based spatial statistical (TBSS) analysis provides a means of assessing alterations in WM tracts. In this paper, both voxel-based morphometry (VBM) and TBSS are examined using DTI data of temporal lobe epilepsy (TLE) patients and nonepileptic subjects. In addition to fractional anisotropy (FA), ellipsoidal area ratio (EAR) is used in this study. Significant reductions of FA and EAR are identified by TBSS in the parahippocampal white matter. Because of methodological differences, TBSS detects more localized abnormalities than VBM, while the EAR is more sensitive to WM alteration than FA. PMID- 21616485 TI - Mental, psychomotor, neurologic, and behavioral outcomes of 2-year-old children born after preimplantation genetic screening: follow-up of a randomized controlled trial. AB - OBJECTIVE: To evaluate the effect of preimplantation genetic screening (PGS) on neurodevelopmental outcomes in children. DESIGN: Prospective, assessor-blinded, follow-up study of children born to women randomly assigned to in vitro fertilization or intracytoplasmic sperm injection (IVF/ICSI) with or without PGS. SETTING: University Medical Center, Groningen, and Academic Medical Center, Amsterdam, the Netherlands. PATIENT(S): Fifty-four PGS children and 77 controls. INTERVENTION(S): PGS. MAIN OUTCOME MEASURE(S): Mental, psychomotor, neurologic, and behavioral outcomes in 2-year-old children as measured with the Bayley Scales of Infant Development, the Hempel neurologic examination, and the Child Behavior Check List. RESULT(S): The mental, psychomotor, and behavioral outcomes at 2 years in children born after IVF with and without PGS were similar overall. The PGS children showed lower neurologic optimality scores than the control children. Scores on all tests were within the normal range. CONCLUSION(S): Conception with PGS does not seem to be associated with impaired mental, psychomotor, or behavioral outcomes by age 2. However, the lower neurologic optimality scores found in the PGS children may signal less favorable long-term neurologic outcomes in PGS children. Our findings stress the need for safety evaluations with new assisted reproductive techniques before large-scale implementation. PMID- 21616486 TI - Vascular endothelial growth factor expression up-regulated by endometrial ischemia in secretory phase plays an important role in endometriosis. AB - Primary human endometrial cells were exposed to hypoxia preconditioning (HPC), HPC-hypoxia, and hypoxia conditions, and then endometrial tissue treated with ischemia preconditioning (IPC) was transplanted onto the chick embryo chorioallantoic membrane to investigate the role of slight ischemia of endometrium in the pathologic process of endometriosis. IPC up-regulated vascular endothelial growth factor expression and decreased apoptosis of endometrial cells, thus facilitating the endometrial fragments' ectopic implantation. PMID- 21616487 TI - Socioeconomic disparities in the use and success of fertility treatments: analysis of data from a prospective cohort in the United States. AB - OBJECTIVE: To determine the effect of income, education, and race on the use and outcomes of infertility care. DESIGN: Prospective cohort. SETTING: Eight community and academic infertility practices. PATIENT(S): Three hundred ninety one women presenting for an infertility evaluation. INTERVENTION(S): Face-to-face and telephone interviews and questionnaires. MAIN OUTCOME MEASURE(S): Use of infertility services and odds of pregnancy. Linear and logistic regression used to assess relationship between racial and socioeconomic characteristics, use of infertility services, and infertility outcomes. RESULT(S): After adjustment for age and demographic and fertility characteristics, college-educated couples (beta = $5,786) and households earning $100,000-$150,000 (beta = $6,465) and >=$150,000 (beta = $8,602) spent significantly more on infertility care than their non college-educated, lower-income counterparts. Higher income and college-educated couples were much more likely to use more cycles of higher-intensity fertility treatment. The increased cost of infertility care was primarily explained by these differences in number and type of infertility treatment. Even after adjustment for these factors and total amount spent on fertility care, having a college degree was associated with persistently higher odds of achieving a pregnancy (OR = 1.9). CONCLUSION(S): Education and household income were independently associated with the amount of money spent on fertility care. This relationship was primarily explained by types and intensity of infertility treatments used. Having at least a college degree was independently associated with improved odds of pregnancy. PMID- 21616488 TI - Reconstruction of radial capitellar fractures using fine-threaded implants (FFS). AB - Fractures of the capitulum humeri are rare and challenging injuries to the elbow and outcome results are limited to case series, often with different fixation devices. This article aims to present 15 patients with a fracture of the capitulum humeri treated with the same implant, fine-threaded Kirschner wires (FFS). Besides range of motion and grip strength, outcome measurements include the following scores: the Mayo Elbow Performance index (MEPI); the American Shoulder and Elbow Surgeons' Elbow Assessment (ASES) Form; the Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire; and the Broberg-Morrey score. Using the modified Bryan and Morrey classification, there were seven type-I injuries, three type-II, two type-III and three type-IV fractures. In addition to fractures of the capitellum, five patients were diagnosed with a complex ligamentous injury and three had suffered radial head fractures. After a minimum follow-up of 12 months, we obtained a range of motion in flexion and extension of 124 degrees and a pro- and supination of 173 degrees . The mean DASH showed 10.84 and the Broberg-Morrey score reached 90.8. ASES and MEPI scores were 91.5 and 90, respectively. The mean grip strength of the injured side was 217.9N versus 236.4N on the healthy side. The use of FFS for capitellar fractures of the humerus achieved good clinical results and these implants can be considered as a treatment option for this fracture entity. PMID- 21616489 TI - The concept of difference and the experience of China-educated nurses working in Australia: a symbolic interactionist exploration. AB - BACKGROUND: The migration of registered nurses as a response to a global nurse shortage has seen a growth in research interest in this area. Much of the research focuses on differences in language and culture which are posed as the attributes of the immigrant nurse. OBJECTIVES: In reporting on an analysis of data drawn from China-educated nurses working in the Australian health care system, this paper explores the social construction of difference and the related intersection of difference and racialisation. DESIGN: A symbolic interactionist approach informed 46 in-depth interviews with 28 China-educated nurses. The method of analysis was initial and focused coding and constant comparison of data. The focus of interpretation was on human action and interaction as contextual and thus structural factors were critical to the analysis. RESULTS: Two levels of meaning were depicted in this study: difference as "you are you and I am I" and difference as "incompetence". Negative meanings were ascribed to difference which in turn legitimised inequality and held the potential to perpetuate racism. CONCLUSIONS: We argue that it is problematic to conceptualise difference as individual attributes. Difference needs to be contextualised and thus explored as a complex and socially constructed concept that in its application has social and political implications for immigrant nurses in Australia and elsewhere. PMID- 21616490 TI - A comparison of 25-hydroxyvitamin D serum levels among those with or without statin-associated myalgias. PMID- 21616491 TI - Genetic and environmental regulation of inflammatory CVD biomarkers Lp-PLA2 and IgM anti-PC. AB - OBJECTIVE: We set out to investigate the relative contribution of genetic and environmental effect on two inflammatory CVD biomarkers; lipoprotein-associated phospholipase A(2) (Lp-PLA(2)) and anti-phosphorylcholine IgM (anti-PC). Their relationships and possible co-regulation with other established CVD biomarkers are also examined. METHODS: Lp-PLA(2) activity (N=1600) and anti-PC (N=2036) levels were measured in elderly Swedish twins. Correlation analyses and heritability estimation were conducted by structural equation modeling. RESULTS: We attribute 0.37 of the variance of Lp-PLA(2) and 0.40 of anti-PC variance to genetic variance. In addition, a bivariate heritability of 0.33, 0.35 and 0.36 could be detected for levels of Lp-PLA(2) together with ApoB, total cholesterol and LDL, respectively. Anti-PC was only weakly related to other biomarkers of CVD, which may suggest a more independent role of anti-PC as a biomarker. CONCLUSIONS: In this large sample, Lp-PLA(2) activity has lower heritability and higher environmental regulation than previously reported. Anti-PC levels are partly influenced by dominance genetics and appear to be regulated independently of more established CVD biomarkers. PMID- 21616492 TI - An unbreakable on-line approach towards sol-gel capillary microextraction. AB - In this work a novel unbreakable sol-gel-based in-tube device for on-line solid phase microextraction (SPME) was developed. The inner surface of a copper tube, intended to be used as a high performance liquid chromatography (HPLC) loop, was electrodeposited by metallic Cu followed by the self assembled monolayers (SAM) of 3-(mercaptopropyl) trimethoxysilane (3MPTMOS). Then, poly (ethyleneglycol) (PEG) was chemically bonded to the -OH sites of the SAM already covering the inner surface of the copper loop using sol-gel technology. The homogeneity and the porous surface structure of the SAM and sol-gel coatings were examined using the scanning electron microscopy (SEM) and adsorption/desorption porosimetry (BET). The prepared loop was used for online in-tube SPME (capillary microextraction) of some selected polycyclic aromatic hydrocarbons (PAHs), as model compounds, from the aquatic media. After extraction, the HPLC mobile phase was used for on-line desorption and elution of the extracted analytes from the loop to the HPLC column. Major parameters affecting the extraction efficiency including the sample flow rate through the copper tube, loading time, desorption time and sample volume were optimized. For investigating the sorbent efficiency, four loops based on the copper tube itself, the copper tube after electrodeposition with Cu and the tubes with the SAMs and SAMs-sol-gel coating were made and compared. The SAMs-sol-gel coated loop clearly shows a prominently lead of at least 20-100 times of higher efficiency. The linearity for the analytes was in the range of 0.01-500 MUg L(-1). Limit of detection (LOD) was in the range of 0.005-0.5 MUg L(-1) and the RSD% values (n=5) were all below 8.3% at the 5 MUg L(-1) level. The developed method was successfully applied to real water samples while the relative recovery percentages obtained for the spiked water samples were from 90 to 104%. The prepared loop exhibited long life time due to its remarkable solvent and mechanical stability. Different solvents such as methanol, acetonitrile and acetone were passed through the loop for many days and it was also used for more than 100 extractions/desorption of the selected analytes and no decrease in the peak areas was observed. PMID- 21616494 TI - Liquid chromatography fractionation with gas chromatography/mass spectrometry and preparative gas chromatography-nuclear magnetic resonance analysis of selected nonylphenol polyethoxylates. AB - Commercial nonylphenol polyethoxylates, designated as NPnEOs, where n is the number of ethoxy groups, comprise a range of ethoxylate groups. According to the starting material nonylphenol, they may also be composed of a complex mix of isomeric nonyl substituents. In order to study more fully the heterogeneity arising from both the ethoxylate and nonyl groups, a mixture of NPnEOs is first fractionated by normal phase liquid chromatography (NPLC) into separate fractions comprising individual ethoxymers, n. Preparative collection of each early elution ethoxymer fraction allows further separation of different isomeric nonyl group components by using analytical gas chromatography/mass spectrometry (GC/MS). The nonyl isomers are not resolved in the NPLC method. The distribution of the isomeric nonyl side chain of different ethoxymers bears close resemblance with each other, and also with the original nonylphenol starting material, although separation efficiency of the nonyl isomers for each ethoxymer decreases with increasing ethoxymer number. Mass spectrometry of the separated isomers display close similarity for presumed equivalent isomers in each fraction, based on elution order of the nonyl isomers. This suggests that each corresponding peak has the same isomer structure. Mass spectra are interpreted based on branching within the nonyl side chain. Preparative GC coupled with MS and nuclear magnetic resonance spectroscopy elucidated the molecular structure of one of the resolved isomers as 4-(1,3-dimethyl-1-propyl-butyl)-phenol diethoxylate. PMID- 21616495 TI - On-chip solid phase extraction and enzyme digestion using cationic PolyE-323 coatings and porous polymer monoliths coupled to electrospray mass spectrometry. AB - We evaluate the compatibility and performance of polymer monolith solid phase extraction beds that incorporate cationic charge, with a polycationic surface coating, PolyE-323, fabricated within microfluidic glass chips. The PolyE-323 is used to reduce protein and peptide adsorption on capillary walls during electrophoresis, and to create anodal flow for electrokinetically driven nano electrospray ionization mass spectrometry. A hydrophobic butyl methacrylate-based monolithic porous polymer was copolymerized with an ionizable monomer, [2 (methacryloyloxy)ethyl] trimethylammonium chloride to form a polymer monolith for solid phase extraction that also sustains anodal electroosmotic flow. Exposure of the PolyE-323 coating to the monolith forming mixture affected the performance of the chip by a minor amount; electrokinetic migration times increased by ~5%, and plate numbers were reduced by an average of 5% for proteins and peptides. 1-mm long on-chip monolithic solid phase extraction columns showed reproducible, linear calibration curves (R(2)=0.9978) between 0.1 and 5 nM BODIPY at fixed preconcentration times, with a capacity of 2.4 pmol or 0.92 mmol/L of monolithic column for cytochrome c. Solution phase on-bed trypsin digestion was conducted by capturing model protein samples onto the monolithic polymer bed. Complete digestion of the proteins was recorded for a 30 min stop flow digestion, with high sequence coverage (88% for cytochrome c and 56% for BSA) and minimal trypsin autodigestion product. The polycationic coating and the polymer monolith materials proved to be compatible with each other, providing a high quality solid phase extraction bed and a robust coating to reduce protein adsorption and generate anodal flow, which is advantageous for electrospray. PMID- 21616496 TI - Determination of binding parameters between lysozyme and its aptamer by frontal analysis continuous microchip electrophoresis (FACMCE). AB - An original and simple methodology based on microchip electrophoresis (MCE) in a continuous frontal analysis mode (named frontal analysis continuous microchip electrophoresis, FACMCE) was developed for the simultaneous determination of the binding parameters, i.e. ligand-site dissociation constant (k(d)) and number of binding sites on the substrate (n). This simultaneous determination was exemplified with the interaction between an aptamer and its target. The selected target is a strongly basic protein, lysozyme, as its quantification is of great interest due to its antimicrobial and allergenic properties. A glass microdevice equipped with a fluorescence detection system was coated with hydroxypropylcellulose, reducing the electroosmotic flow and adsorption onto the channel walls. This microdevice allowed the continuous electrokinetic injection of a mixture of fluorescently labelled aptamer and non-labelled lysozyme. By determining the concentration of the free fluorescently labelled aptamer thanks to its corresponding plateau height, mathematical linearization methods allowed to determine a k(d) value of 48.4+/-8.0 nM, consistent with reported results (31 nM), while the average number of binding sites n on lysozyme, never determined before, was 0.16+/-0.03. These results seem to indicate that the buffer nature and the SELEX process should influence the number and affinity of the binding sites. In parallel it has been shown that the binding between lysozyme and its aptamer presents two sites of different binding affinities. PMID- 21616497 TI - Improved partition efficiency with threaded cylindrical column in vortex counter current chromatography. AB - Type-I coil planet centrifuge produces a uniformly circulating centrifugal force field to produce vortex motion of two immiscible solvent phases in a cylindrical cavity of the separation column to perform efficient countercurrent chromatography. The partition efficiency obtained from the original vortex column was substantially improved by threading the cylindrical cavity to increase the area of mass transfer between the two phases. Partition efficiency of the threaded column was evaluated by three different two-phase solvent systems with a broad range of hydrophobicity each with a set of suitable test samples. Overall results of the present studies indicated that the threaded cylindrical column substantially improves the partition efficiency in terms of theoretical plate number, peak resolution, and height equivalent of one theoretical plate. The results also indicated that higher peak resolution is produced by eluting either the upper phase in the head to tail direction or the lower phase in the reversed direction. When there is a choice in the mobile phase, a better separation is achieved by using the less viscous phase as the mobile phase. Since the present system gives extremely low column pressure, it may be a potential alternative to the conventional type-J HSCCC system for a large-scale preparative separation. PMID- 21616498 TI - Determination of per- and polyfluorinated substances in airborne particulate matter by microwave-assisted extraction and liquid chromatography-tandem mass spectrometry. AB - A sensitive and confirmatory analytical method has been developed for the determination of 12 ionic per- and polyfluorinated alkyl substances (PFAS) in fine airborne particulate matter (PM2.5) at trace levels. The proposed method includes extraction of PM2.5-bound PFAS by microwave-assisted extraction (MAE) followed by centrifugation and injection into the liquid chromatograph coupled to a triple quadrupole tandem mass spectrometry system (LC-MS/MS). The main parameters affecting the performance of MAE were optimised using statistical design of experiments (DoE). Recoveries ranged from 83 to 120% and the method quantification limit (MQL) was 1.4 pg m(-3), when air volumes of 720 m(3) were sampled. This method was successfully applied to 41 samples collected from five stations of the monitoring network of the Valencian Regional Government (Spain) during April-July 2010. Eight out of 12 PFCs investigated were quantified in at least one sample (PFBA, PFPeA, PFHxS, 6:2 FTS, PFOA, PFNA, PFOS and PFDA). The measured concentrations ranged from 1.4 to 34.3 pg m(-3). PMID- 21616499 TI - High spatial resolution label-free detection of antigen-antibody binding on patterned surface by imaging ellipsometry. AB - High spatial resolution and large area thickness mapping of label-free protein microarray has been achieved using imaging ellipsometry (IE) under optimized conditions. The protein patterns with feature size down to 8*8 MUm(2) was readily imaged, and the binding between the surface immobilized antigen and the antibody was monitored. Quantitative thickness analysis of antibody-antigen binding on the 32*32 MUm(2) micron spots was successfully performed, and we have obtained a limit of detection as low as 1.2 pg/spot. This work demonstrates that appropriately optimized IE could be used as a highly sensitive and high through put label-free technique for studying surface antigen-antibody recognition in sub 40 MUm scale. PMID- 21616500 TI - Moderating effect of ammonia on particle growth and stability of quasi monodisperse silver nanoparticles synthesized by the Turkevich method. AB - A new method to stabilize silver nanoparticles by the addition of ammonia is proposed. Colloidal dispersions of silver nanoparticles were synthesized by the Turkevich method using sodium citrate to reduce silver nitrate at high pH and at 90 degrees C. After approximately 12 min, a diluted ammonia solution was added to the reaction flask to form soluble diamine silver (I) complexes that played an important growth moderating role, making it possible to stabilize metallic silver nanoparticles with sizes as small as 1.6 nm after 17 min of reaction. Colloidal dispersions were characterized by UV-visible absorption spectroscopy, X-ray diffraction, and transmission electronic microscopy. PMID- 21616501 TI - CdS nanoparticles deposited on montmorillonite: preparation, characterization and application for photoreduction of carbon dioxide. AB - CdS nanoparticles were precipitated by the reaction of cadmium acetate with sodium sulphide in the presence of cetyltrimethylammonium (CTA) and deposited on montmorillonite (MMT). The resulting CdS-MMT nanocomposite contained 6 wt.% of CdS and 30 wt.% of CTA. Band-gap energy of CdS was estimated at 2.63+/-0.09 eV using the Tauc plot. The size of CdS nanoparticles was calculated from the band gap energy at 5 nm and from the micrographs of transmission electron microscopy (TEM) at 5 nm. Selected area electron diffraction (SAED) recognized the cubic structure of CdS (Hawleite). The dynamic light scattering (DLS) method confirmed that CdS nanoparticles were anchored on the surface of MMT particles. CTA was found to be intercalated into MMT and adsorbed on its external surface. CdS-MMT was used for the photoreduction of carbon dioxide dissolved in NaOH solutions. The yields of originating gas products can be arranged in the order: H(2) ? CH(4) > CO. Amounts of these products were 4-8 folds higher then those obtained with TiO(2) Evonic P25. Hydrogen reduced CO(2) to CO and CH(4). PMID- 21616503 TI - The need for phosphoproteomic approaches in psychiatric research. PMID- 21616502 TI - Predicting sights from sounds: 6-month-olds' intermodal numerical abilities. AB - Although the psychophysics of infants' nonsymbolic number representations have been well studied, less is known about other characteristics of the approximate number system (ANS) in young children. Here three experiments explored the extent to which the ANS yields abstract representations by testing infants' ability to transfer approximate number representations across sensory modalities. These experiments showed that 6-month-olds matched the approximate number of sounds they heard to the approximate number of sights they saw, looking longer at visual arrays that numerically mismatched a previously heard auditory sequence. This looking preference was observed when sights and sounds mismatched by 1:3 and 1:2 ratios but not by a 2:3 ratio. These findings suggest that infants can compare numerical information obtained in different modalities using representations stored in memory. Furthermore, the acuity of 6-month-olds' comparisons of intermodal numerical sequences appears to parallel that of their comparisons of unimodal sequences. PMID- 21616504 TI - Multiple acyl-CoA-dehydrogenase deficiency (MADD)--a novel mutation of electron transferring-flavoprotein dehydrogenase ETFDH. AB - This is the case of a 41 year old man, suffering general weakness and elevated liver enzymes, sensitive to a treatment with riboflavin and coenzyme Q(10). Tandem mass spectroscopy and molecular analysis reveal a multiple acyl-CoA dehydrogenase deficiency (MADD) with two novel heterozygote missense mutations of the EFTDH gene. PMID- 21616505 TI - NOTCH3 gene mutations in subjects clinically suspected of CADASIL. AB - BACKGROUND: Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is an inherited cerebrovascular disease due to mutations involving loss or gain of a cysteine residue in the NOTCH3 gene. A cluster of mutations around exons 3 and 4 was originally reported. Identification of pathogenic mutation is important for diagnostic confirmation of the disease, however genetic counselling and testing of relatives at risk is critical in mutation carriers. METHODS: Mutation analysis of the NOTCH3 gene was performed through direct sequencing in 140 patients with clinical suspicion of CADASIL. Patients underwent genetic counselling pre and post testing. The 2-23 exons containing all EGF-like domains were screened. RESULTS: 14 familial forms of the disease have been identified with 14 different causative mutations in exons 2, 3, 4, 5, 7, 10, 14, 19, 20 and 22 of the NOTCH3 gene; no pathogenetic mutations have been identified in exons 6 and 8; several genetic variations both in coding as well as in intronic regions were identified too. CONCLUSIONS: Our data confirm the importance of screening the whole EGF-like domains region of NOTCH3 gene for the molecular diagnosis of CADASIL among the Italian population too. Moreover genetic variants different from loss or gain of a cysteine residue are identified and presented. PMID- 21616506 TI - Determination of the elastic modulus of ascending thoracic aortic aneurysm at different ranges of pressure using uniaxial tensile testing. AB - OBJECTIVE: The purpose of this study is to provide measurements of the elastic modulus of the aortic wall of ascending thoracic aortic aneurysms for different ranges of pressure (physiologic, hypertensive). In addition, pre-failure stress, taken as the peak stress obtained before specimen failure, was recorded for each test. METHODS: Ninety-seven aortic samples freshly excised from 13 patients with ascending thoracic aortic aneurysms were obtained from greater and lesser curvatures and tested uniaxially in circumferential and longitudinal orientations. RESULTS: The maximum elastic moduli, overall, and particularly in the lesser curvature were significantly higher in the circumferential orientation (9.19 MPa) than in the longitudinal (3.13 MPa). Results of peak stress showed positive correlation with maximum elastic modulus and inverse correlation with tissue wall thickness. CONCLUSIONS: This study provides new data on the elastic modulus in the physiologic and hypertensive range that can be used in computational analysis and the design of bench-top models. The accuracy of computational analysis and bench-top models strongly depends on the knowledge of the elastic properties of the aortic wall. The mechanical properties presented in this study, with specific values for 2 locations (greater and lesser curvature) and 2 directions (circumferential, longitudinal), will increase our understanding of the mechanisms that precede rupture of an ascending aortic aneurysm. PMID- 21616507 TI - Designing comparative effectiveness trials of surgical ablation for atrial fibrillation: experience of the Cardiothoracic Surgical Trials Network. AB - OBJECTIVE: Since the introduction of the cut-and-sew Cox maze procedure for atrial fibrillation, there has been substantial innovation in techniques for ablation. Use of alternative energy sources for ablation simplified the procedure and has resulted in dramatic increase in the number of patients with atrial fibrillation treated by surgical ablation. Despite its increasingly widespread adoption, there is lack of rigorous clinical evidence to establish this procedure as an effective clinical therapy. METHODS: This article describes a comparative effectiveness randomized trial, supported by the Cardiothoracic Surgical Clinical Trials Network, of surgical ablation with left atrial appendage closure versus left atrial appendage closure alone in patients with persistent and long-standing persistent atrial fibrillation undergoing mitral valve surgery. Nested within this trial is a further randomized comparison of 2 different lesions sets: pulmonary vein isolation and the full maze lesion set. RESULTS: This article addresses trial design challenges, including how best to characterize the target population, operationalize freedom from atrial fibrillation as a primary end point, account for the impact of antiarrhythmic drugs, and measure and analyze secondary end points, such as postoperative atrial fibrillation load. CONCLUSIONS: This article concludes by discussing how insights that emerge from this trial may affect surgical practice and guide future research in this area. PMID- 21616508 TI - Spinal cord injury in patients undergoing total arch replacement: a cautionary note for use of the long elephant technique. AB - OBJECTIVES: Spinal cord injury in patients undergoing total arch replacement via median sternotomy is a rare but catastrophic complication. With recent surgical advances, including the long elephant trunk technique or open stent procedure, the incidence of spinal cord injury is increasing. In our institute, the long elephant trunk procedure is routinely performed when appropriate to the patient's status. The aim of our study was to determine risk factors for spinal cord injury during total arch replacement. METHODS: Since April 2004, 61 patients underwent total arch replacement with a 4-branched prosthetic graft. The study population included 44 men and 17 women (mean age, 69.8 +/- 11.1 years). Thirty-three patients underwent the long elephant trunk procedure with a mean length of 12.0 +/- 3.5 cm (range, 7-21.6 cm). The Adamkiewicz artery was detected by computed tomography in 11 patients. We assessed the perioperative factors associated with spinal cord injury. RESULTS: Four (6.6%) patients had spinal cord injury (3 cases of paraplegia and 1 case of paraparesis). This constitutes 12% of all patients having the long elephant trunk procedure. No patients with direct distal anastomosis had a spinal cord injury. Univariate logistic regression analysis identified female gender, elephant trunk more than 10 cm from the left subclavian artery, and nonpreserved Adamkiewicz artery as significant independent risk factors for spinal cord injury. Both female gender (P = .017) and long elephant trunk (P = .005) were significant by multivariate analysis. CONCLUSIONS: Elephant trunk more than 10 cm from the left subclavian artery was associated with increased risk of spinal cord injury. We recommend short elephant trunk or long elephant trunk with preservation of the Adamkiewicz artery to prevent spinal cord injury in patients having total arch replacement. PMID- 21616509 TI - Plastic ingestion in Franciscana dolphins, Pontoporia blainvillei (Gervais and d'Orbigny, 1844), from Argentina. AB - Plastic debris (PD) ingestion was examined in 106 Franciscana dolphins (Pontoporia blainvillei) incidentally captured in artisanal fisheries of the northern coast of Argentina. Twenty-eight percent of the dolphins presented PD in their stomach, but no ulcerations or obstructions were recorded in the digestive tracts. PD ingestion was more frequent in estuarine (34.6%) than in marine (19.2%) environments, but the type of debris was similar. Packaging debris (cellophane, bags, and bands) was found in 64.3% of the dolphins, with a lesser proportion (35.7%) ingesting fishery gear fragments (monofilament lines, ropes, and nets) or of unknown sources (25.0%). PD ingestion correlated with ontogenetic changes in feeding regimes, reaching maximum values in recently weaned dolphins. Because a simultaneous increase in gillnet entanglement and the bioaccumulation of heavy metals take place at this stage, the first months after trophic independence should be considered as a key phase for the conservation of Franciscana dolphin stocks in northern Argentina. PMID- 21616510 TI - Distribution, origin, and potential toxicological significance of polycyclic aromatic hydrocarbons (PAHs) in sediments of Kaohsiung Harbor, Taiwan. AB - Sediment samples were collected from 12 locations of Kaohsiung Harbor, Taiwan and analyzed for polycyclic aromatic hydrocarbons (PAHs). Total PAH concentrations varied from 472 to 16,201 ng g(-1) dry weight. The highest PAH concentrations were from the industrial zone docks situated in south Kaohsiung Harbor. Diagnostic ratios showed that the possible source of PAHs in the industrial zone dock could be coal combustion while in the other zones it could be petroleum combustion. The toxic equivalent concentrations (TEQcarc) of PAHs varied from 55 to 1964 ng TEQ g(-1) dry weight. Higher total TEQcarc values were found at industrial zone docks (from 1404 to 1964 ng TEQ g(-1) dry weight). As compared with the US Sediment Quality Guidelines (SQGs), the observed levels of PAHs at industrial zone docks exceeded the effects range low (ERL), and could thus cause acute biological damage. However, the lower levels of PAHs at the other zones would probably not exert adverse biological effects. PMID- 21616511 TI - Cetaceans and military sonar: a need for better management. PMID- 21616512 TI - High-dose insulin administration is associated with hypoaminoacidemia during cardiac surgery. AB - Although the effects of insulin on glucose homeostasis are well recognized in surgical patients, its effect on perioperative protein metabolism has received little attention. The purpose of this study was to examine the effect of high dose insulin therapy on the plasma concentrations of amino acids (AAs) in patients undergoing coronary artery bypass grafting surgery. We studied 20 nondiabetic patients scheduled for elective coronary artery bypass grafting surgery. Patients were randomly allocated to receive either standard metabolic care (target glycemia 6.0-10.0 mmol/L, control group, n = 10) or high-dose insulin therapy (insulin group, n = 10). Insulin was administered at 5 mU.kg( 1).min(-1) beginning at skin incision. Simultaneously, 20% dextrose was infused at a variable rate adjusted to maintain glycemia between 4.0 and 6.0 mmol/L. Plasma AAs, glucose, cortisol, and insulin were measured immediately before surgery and at sternal closure. Differences in mean values were assessed by Student t test. Plasma concentrations of all AAs decreased in the insulin group, with 15 of 22 AAs, including all branched-chain AAs, being significantly lower at sternal closure when compared with the control group. At the end of surgery, plasma glucose concentration was significantly lower in the insulin group (4.2 +/ 0.6 vs 7.3 +/- 1.0 mmol/L, P = .0001), whereas plasma cortisol levels did not show any difference between groups. High-dose insulin therapy resulted in a significant reduction in plasma AAs, particularly branched-chain AAs, during cardiac surgery. PMID- 21616513 TI - A population-based study on incidence and economic burden of influenza-like illness in south China, 2007. AB - OBJECTIVES: The disease burden of influenza-like illness (ILI) in most tropical and subtropical countries has not been described adequately to date. The aim of this study was to determine the epidemiology and incidence of ILI, and to assess the economic burden in south China. STUDY DESIGN: Prospective study. METHODS: A population-based household survey was conducted quarterly in 2007 in Guangdong Province. RESULTS: The average number of subjects in each quarterly survey was 13,687. In total, 1002 cases of self-reported ILI were identified in all four surveys, indicating an annual incidence of 7.23 cases of ILI per 100 persons. The second quarter had the highest incidence of ILI (2.83 cases per 100 persons). Children aged 1-4 years, <1 year and 5-9 years had the highest annual incidence rates of ILI (49.87, 35.19 and 21.24 cases per 100 persons, respectively). The incidence of ILI was significantly higher in males than females (P < 0.001), and significantly higher in rural residents than urban residents (P < 0.001). The individual cost per episode of ILI represented approximately 20% of monthly per capita income of residents. CONCLUSIONS: The results of this large-scale household study confirm that ILI places a substantial health and economic burden on south China. Ultimately, the results of this study will provide further information for understanding the disease burden of influenza in subtropical areas. PMID- 21616514 TI - Socio-economic aspects of improved sanitation in slums: a review. AB - This socio-economic review provides an overview of the sanitation crisis in slum areas, and re-emphasizes the importance of sanitation. It highlights a lack of recognition of actual drivers for sanitation improvements, and the complexities in the provision of sanitation services in the context of urban slums with a mix of tenants and landlords. It elaborates how the drivers of demand for sanitation outlined in contemporary research are not universal but are rather context specific. The authors point out specific knowledge gaps for future research; for example, the need to establish a scientific basis for context-specific drivers of demand for sanitation improvements in slums, and a better understanding of associated complexities in order to set boundary conditions for achieving desired improvements. PMID- 21616515 TI - [An anterior maxillary swelling]. PMID- 21616516 TI - Direct agglutination test (DAT): improvement of biosafety for laboratory diagnosis of visceral leishmaniasis. AB - In this study, the direct agglutination test (DAT), using 2-mercaptoethanol (2 ME), kaolin or N-acetylcysteine (NAC) as sample diluents, was used to assay 89 samples from visceral leishmaniasis (VL) patients and 130 samples from patients with other diseases and healthy individuals. Maintaining a cut-off of 1:100, the DAT assays with 2-ME, kaolin or NAC presented sensitivities of 94.4%, 95.5% and 100% (P = 0.09) and specificities of 99.2%, 100% and 97.7% (P = 0.17), respectively. Based on these results, we suggest that NAC can be used as a replacement for 2-ME in the DAT, increasing biosafety in the diagnosis of VL. PMID- 21616517 TI - The role of p53 in the response of tumor cells to sonodynamic therapy in vitro. AB - p53 plays a pivotal role in apoptosis. In addition, p53 is currently extensively investigated as a promising strategy for highly specific anticancer therapy in chemotherapeutics and photodynamic therapy. However, the role of p53 in the response of tumor cells to sonodynamic therapy treatment is still unclear. In this study, we aim to investigate the activation of p53 in sonodynamic therapy. Three murine tumor models with distinct aggressiveness (S180, H-22 and EAC) were treated with 1.75MHz continuous ultrasound at an acoustic intensity (I(SATA)) of 1.4W for 3min in the presence of 20MUg/ml hematoporphyrin. The DNA fragment and nuclear damage were observed by TUNEL and single cell gel electrophoresis. Western blotting and RT-PCR were used to analyze the expression of p53, PUMA, Bax and Fas. Then we checked the translocation of p53 by confocal microscopy. DNA sequencing was used to determine the status of p53 gene in three tumor cell lines. Our results indicated that the level of p53 protein and mRNA increased significantly, and p53 activated the expression of its downstream pro-apoptosis gene PUMA, Bax and Fas in the S180 and H-22 cells. Meanwhile, p53 protein translocated onto mitochondria. In the EAC cells, expression and translocation of p53 was not found; the level of PUMA, Bax and Fas remained unaltered. The S180 cells showed most serious DNA fragment and nuclear damage with 77.43% TDNA; H-22 cells in the middle with 58.85% TDNA; whereas EAC cells appeared less nuclear material lost with just 15.82% TDNA. The results of DNA sequencing showed that the sequences of exons 5-8 of the p53 gene of S180, H-22 and EAC cells were the same with the sequences of wild-type p53 provided by NCBI. These results primarily demonstrated that: (1) p53 was activated to promote SDT-induced apoptosis through extrinsic and intrinsic signaling pathways in the S180 and H-22 cells; (2) cellular responses of different cells to SDT were distinct, the aggressive S180 cells were much more sensitive than H-22, whereas EAC cells were relatively less sensitive. The discrepancy among the cell lines may be due to different activation time of p53 protein. PMID- 21616518 TI - Biologically induced phosphorus precipitation in aerobic granular sludge process. AB - Aerobic granular sludge is a promising process for nutrient removal in wastewater treatment. In this work, for the first time, biologically induced precipitation of phosphorus as hydroxyl-apatite (Ca(5)(PO(4))(3)(OH)) in the core of granules is demonstrated by direct spectral and optical analysis: Raman spectroscopy, Energy dispersive X-ray (EDX) coupled with Scanning Electron Microscopy (SEM), and X-ray diffraction analysis are performed simultaneously on aerobic granules cultivated in a batch airlift reactor for 500 days. Results reveal the presence of mineral clusters in the core of granules, concentrating all the calcium and considerable amounts of phosphorus. Hydroxyapatite appears as the major mineral, whereas other minor minerals could be transiently produced but not appreciably accumulated. Biologically induced precipitation was responsible for 45% of the overall P removal in the operating conditions tested, with pH varying from 7.8 to 8.8. Major factors influencing this phenomenon (pH, anaerobic phosphate release, nitrification denitrification) need to be investigated as it is an interesting way to immobilize phosphorus in a stable and valuable product. PMID- 21616519 TI - Spatial distribution of organochlorine contaminants in soil, sediment, and fish in Bikini and Enewetak Atolls of the Marshall Islands, Pacific Ocean. AB - Several nuclear tests were performed at Enewetak and Bikini Atolls in the Marshall Islands between 1946 and 1958. The events at Bikini Atoll involved several ships that were tested for durability during nuclear explosions, and 24 vessels now rest on the bottom of the Bikini lagoon. Nine soil samples were collected from different areas on the two islands of the atoll, and eighteen sediment, nine fish, and one lobster were collected in the vicinity of the sunken ships. Organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), and polychlorinated terphenyls (PCTs) in these samples were analyzed using gas chromatography/ion trap mass spectrometry (GC/ITMS). The average recoveries ranged from 78% to 104% for the different PCB congeners. The limits of detection (LOD) for PCBs, PCTs, DDE, DDT, and dieldrin ranged 10-50 pg g(-1). Some fish from Enewetak contained PCBs at a concentration range of 37-137 ng g(-1), dry weight (dw), and most of the soils from Enewetak showed evidence of PCBs (22-392 ng g(-1)dw). Most of the Bikini lagoon sediment samples contained PCBs, and the highest was the one collected from around the Saratoga, an aircraft carrier (1555 ng g(-1)dw). Some of the fish samples, most of the soil samples, and only one of the sediment samples contained 2,2-bis(4-chlorophenyl)-1,1-dichloroethylene (DDE) and PCBs. In addition to PCBs, the soils from Enewetak Atoll contained PCTs. PCTs were not detected in the sediment samples from Bikini Atoll. The results suggest local pollution sources of PCBs, PCTs, and OCPs. PMID- 21616520 TI - Clinical efficacy and safety of denosumab in postmenopausal women with low bone mineral density and osteoporosis: a meta-analysis. AB - OBJECTIVE: Clinical trials indicate that denosumab could be a potential treatment for postmenopausal osteoporosis. The objective of this meta-analysis was to assess the clinical efficacy and safety of offering denosumab to postmenopausal women with low bone mass. METHODS: Data sources included MEDLINE, EMBASE, and Cochrane Central Register of Controlled Trials (CENTRAL) from inception to February 3, 2010 and bibliographies of reviews. Randomized controlled trials comparing the efficacy and safety of denosumab to placebo for treatment of low bone mass (low bone mineral density or osteoporosis) in postmenopausal women were selected. Two reviewers independently abstracted data on study general characteristics and outcomes. Review Manager 5.0 software was used for data syntheses and meta-analysis. RESULTS: The database search revealed 4 studies (comprising 8864 patients randomized) that met the inclusion criteria and contributed to some or all of the meta-analysis outcomes. Relative risk (95% CI) of fractures for the denosumab compared with placebo group was 0.58 (0.52 to 0.66); relative risk (95% CI) of serious adverse events was 1.33 (0.83 to 2.14); relative risk (95% CI) of serious adverse events related to infection was 2.10 (0.64 to 6.90); relative risk (95% CI) of neoplasm was 1.11 (0.91 to 1.36); relative risk (95% CI) of study discontinuation due to adverse events was 1.10 (0.83 to 1.47); and relative risk (95% CI) of death was 0.78 (0.57 to 1.06). Findings remained robust to sensitivity analyses. CONCLUSION: Our analysis found a significant reduction in relative fracture risk in the denosumab compared with the placebo group. PMID- 21616521 TI - Sonographic evidence of synovitis as a predictor of response to corticosteroid injection in osteoarthritis: comment on the article by Hayashi et al. PMID- 21616522 TI - Expression of indoleamine 2,3-dioxygenase predicts shorter survival in patients with vulvar squamous cell carcinoma (vSCC) not influencing on the recruitment of FOXP3-expressing regulatory T cells in cancer nests. AB - OBJECTIVE: Regulatory T cells (Tregs), and the enzyme indoleamine 2,3-dioxygenase (IDO), have potential regulatory properties for immune escape in cancer. Inhibitors of IDO are available and could potentially be used in vulvar cancer if IDO was proved to drive progression of the disease. The aim of this study was to evaluate the expression of factor forkhead boxP3 (FOXP3), a marker of Tregs, and IDO in vulvar squamous cell carcinoma (vSCC), and to verify their prognostic significance. METHODS: 76 primary tumors and 35 lymph node metastases derived from 76 patients with full clinical history were analyzed. The intratumoral infiltration of Tregs and IDO expression within cancer were evaluated by immunohistochemistry. RESULTS: The number of Tregs in primary tumor and in corresponding lymph node metastasis was significantly correlated. Intensity of Treg infiltrates in the primary and metastatic sites was not correlated to IDO expression and had no influence on the overall patient survival. High IDO expression was associated with significantly worse overall survival among vSCC patients and was found to be an independent prognostic factor similarly to the tumor grade and patient's age. CONCLUSIONS: The degree of intratumoral Treg infiltrates is an individual feature and remains stable throughout the course of the disease without impact on the patient's survival. IDO expression predicts shorter survival of vSCC patients. If immunologic tolerance of the tumor is promoted by the overexpression of IDO it will not influence the number of intratumoral Tregs. IDO expression seems to be an independent prognostic factor in patients with vSCC. PMID- 21616523 TI - Xanthohumol decreases Notch1 expression and cell growth by cell cycle arrest and induction of apoptosis in epithelial ovarian cancer cell lines. AB - OBJECTIVE: Notch1 signaling is active in ovarian cancer and is a promising pathway for new therapies in ovarian cancer. We have previously detected high Notch1 expression in ovarian tumors. Xanthohumol has been shown to inhibit cancer cell growth and invasion, including Kaposi's sarcoma, which also highly expresses Notch1. We hypothesized that the Notch1 signaling pathway is targeted by xanthohumol leading to decreased ovarian cancer cell growth. METHODS: SKOV3 and OVCAR3 cells were utilized. MTT growth assays were conducted following treatment with xanthohumol. Quantitative RT-PCR and Western blot analyses were conducted to assess Notch1 down-regulation. Luciferase reporter assays were performed to assess functional down-regulation of Notch1. Cell cycle analysis was performed by flow cytometry. RESULTS: Significant growth inhibition and down-regulation of Notch1 transcription and protein expression were found following xanthohumol treatment. In addition, xanthohumol increased Hes6 transcription and decreased Hes1 transcription, known downstream targets of Notch 1. These observations were associated with cell cycle inhibition as demonstrated by an increase in p21 expression and S and G2/M cell cycle arrest confirmed by an increase in phosphorylated cdc2. Furthermore, an increase in the apoptotic markers, cleaved caspase-3 and cleaved PARP were observed. CONCLUSION: Xanthohumol was a potent inhibitor of ovarian cancer cell growth, and our results suggest that xanthohumol may be influencing the Notch1 pathway. These findings suggest that xanthohumol could be useful as a therapeutic agent in ovarian cancer. PMID- 21616524 TI - Let-7 microRNA-mediated regulation of IL-13 and allergic airway inflammation. AB - BACKGROUND: IL-13, a cytokine secreted by T(H)2 lymphocytes and other cells, critically modulates allergic inflammation and tissue remodeling in allergic asthma. Although much is known about transcriptional regulation of IL-13, posttranscriptional regulation is poorly understood. OBJECTIVE: Because many inflammatory pathways are known to be regulated by microRNAs, permitting a rapid and fine-tuned response, the role of microRNA-mediated regulation of IL-13 was investigated using both in vitro and in vivo studies. METHODS: A combination of in silico approaches and in vitro transfections in A549 cells and primary cultured T cells was used to demonstrate the involvement of let-7 in IL-13 regulation. Furthermore, intranasal delivery of let-7 microRNA mimic in mice was performed to study its effects in allergic airway inflammatory conditions. RESULTS: Using a combination of bioinformatics and molecular approaches, we demonstrate that the let-7 family of microRNAs regulates IL-13 expression. Induced levels of IL-13 in cultured T cells were inversely related to let-7 levels. In an IL-13-dependent murine model of allergic airway inflammation, we observed that inflammation was associated with a reduction in most of the members of the let-7 family. Exogenous administration of let-7 mimic to lungs of mice with allergic inflammation resulted in a decrease in IL-13 levels, resolution of airway inflammation, reduction in airway hyperresponsiveness, and attenuation of mucus metaplasia and subepithelial fibrosis. CONCLUSION: Let-7 microRNAs inhibit IL-13 expression and represent a major regulatory mechanism for modulating IL-13 secretion in IL-13-producing cell types and thereby T(H)2 inflammation. PMID- 21616525 TI - Treatment with propranolol of 6 patients with idiopathic aquagenic pruritus. PMID- 21616526 TI - Vascular events after stroke: terutroban fails to PERFORM. PMID- 21616528 TI - The treatment of Glioblastoma Xenografts by surfactant conjugated dendritic nanoconjugates. AB - Polysorbate 80 (P80) anchored poly(propyleneimine) (PPI) dendritic nanoconjugate was developed and evaluated for targeting anti-cancer drug, docetaxel (DTX) to the brain tumor. In vitro cytotoxicity studies of free DTX, DTX-PPI and DTX-P80 PPI dendrimers were carried out using U87MG human glioblastoma cell line. The in vivo anti-cancer activity in brain tumor bearing rats revealed that DTX loaded P80 conjugated dendrimers reduced the tumor volume extremely significantly (p < 0.0001; more than 50%). The median survival time for brain tumor bearing rats treated with DTX-P80-PPI dendrimers (42 days) was extended very significantly as compared to DTX-PPI (23 days; p < 0.001), receptor blocked group (15 days; p < 0.001) and free DTX (18 days; p < 0.001). Gamma scintigraphy and biodistribution studies further confirmed the targeting efficiency and higher biodistribution of ligand conjugated dendrimer into the brain. The results concluded that the developed nanoconjugate has potential to deliver significantly higher amount of drug to brain tumor for improved therapeutic outcome. PMID- 21616529 TI - Near-infrared light induced in vivo photodynamic therapy of cancer based on upconversion nanoparticles. AB - Upconversion nanoparticles (UCNPs) that emit high-energy photons upon excitation by the low-energy near-infrared (NIR) light are emerging as new optical nano probes useful in biomedicine. Herein, we load Chlorin e6 (Ce6), a photosensitizer, on polymer-coated UCNPs, forming a UCNP-Ce6 supramolecular complex that produces singlet oxygen to kill cancer cells under NIR light. Excellent photodynamic therapy (PDT) efficacy is achieved in tumor-bearing mice upon intratumoral injection of UCNP-Ce6 and the followed NIR light exposure. It is further uncovered that UCNPs after PDT treatment are gradually cleared out from mouse organs, without rendering appreciable toxicity to the treated animals. Moreover, we demonstrate that the NIR-induced PDT based on UCNP-Ce6 exhibits a remarkably increased tissue penetration depth compared to the traditional PDT using visible excitation light, offering significantly improved treatment efficacy for tumors blocked by thick biological tissues. Our work demonstrates NIR light-induced in vivo PDT treatment of cancer in animals, and highlights the promise of UCNPs for multifunctional in vivo cancer treatment and imaging. PMID- 21616530 TI - Regenerative potentials of platelet-rich plasma enhanced by collagen in retrieving pro-inflammatory cytokine-inhibited chondrogenesis. AB - This study was undertaken to evaluate the role of collagen matrix to enhance platelet-rich plasma (PRP) effects on pro-inflammatory cytokine-induced arthritic model. We have previously demonstrated the highly regenerative roles of PRP to restore disc degeneration and osteoporosis. In this study, PRP modulated by collagen matrix was used as a regenerative and anti-inflammatory mediator to rescue the chondrocyte degeneration induced by pro-inflammatory cytokines IL 1beta (10 ng/ml)+TNF-alpha (20 ng/ml). First, the MTT result indicated that 1 ng/ml TGF-beta1 in PRP showed an optimal dosage for chondrocytes proliferation. The chondrogenic-specific gene expressions were rescued by PRP from the inhibition of IL-1beta+TNF-alpha, especially under the modulation of collagen matrix. The inflammatory molecules activated by IL-1beta+TNF-alpha were also significantly diminished by PRP with collagen matrix. The membrane receptors integrin alpha1beta1 and CD44 were strongly inhibited by IL-1beta+TNF-alpha, while this inhibition was then recovered by PRP in collagen coating condition. In a 3D model encapsulated with collagen, PRP-induced chondrogenesis were highly enhanced, such as strong restoration of type II collagen and proteoglycan from the inhibition of IL-1beta+TNF-alpha. The result indicated that collagen matrix enhances the effect of PRP on chondrogenesis in response to pro-inflammatory cytokines. The combination of PRP and collagen matrix might facilitate a physiological microenvironment beneficial for maintaining chondrocyte homeostasis and represents an advanced osteoarthritis therapy for clinical applications. PMID- 21616531 TI - STIM and Orai in platelet function. AB - Physiological platelet activation and thrombus formation are essential to stop bleeding in case of vascular injury, whereas inadequate triggering of the same process in diseased vessels can lead to fatal thromboembolism and tissue ischemia of vital organs. A central step in platelet activation is agonist-induced elevation of the intracellular Ca(2+) concentration. This happens on the one hand through the release of Ca(2+) from intracellular stores and on the other hand through Ca(2+) influx from the extracellular space. In platelets, the major Ca(2+) influx pathway is the so-called store operated Ca(2+) entry (SOCE), induced by store depletion. Studies in the last five years discovered the molecular background of platelet SOCE. Stromal interaction molecule 1 (STIM1) and Orai1, two so far unknown molecules, got in the focus of research. STIM1 was found to be the Ca(2+) sensor in the endoplasmic reticulum (ER) membrane, whereas Orai1 was identified as the major store operated Ca(2+) (SOC) channel in the plasma membrane. These two molecules and their role in platelet function and thrombus formation are the topic of the present review with a special focus on apoptosis and apoptosis-like processes in platelet physiology. PMID- 21616532 TI - A menage a trois made in heaven: G-protein-coupled receptors, lipids and TRP channels. AB - Recent technical and conceptual advances in lipid analysis have given us a glimpse into the true versatility of the lipidome and the complexity of lipid signaling species. Progress alike in protein chemistry and genetics has presented us with new signal pathways and molecular mechanisms for the lipid actions. G protein-coupled receptors (GPCR) appear to play a central role in the regulation of many lipid signals and are also themselves targets for some of these. TRP channels have recently been acknowledged as one of the most important GPCR effectors; in many cases the signals from GPCRs to TRPs are mediated via lipid signals. This review aims at presenting a view into the complex lipid signaling networks, their possible regulation by GPCRs and the signals transmitted to the TRP channels. Critical views and possible shortcomings in the composition of the studies are also presented. PMID- 21616533 TI - Enhanced ER Ca2+ store filling by overexpression of SERCA2b promotes IP3-evoked puffs. AB - Liberation of Ca(2+) from the endoplasmic reticulum (ER) through inositol trisphosphate receptors (IP(3)R) is modulated by the ER Ca(2+) content, and overexpression of SERCA2b to accelerate Ca(2+) sequestration into the ER has been shown to potentiate the frequency and amplitude of IP(3)-evoked Ca(2+) waves in Xenopus oocytes. Here, we examined the effects of SERCA overexpression on the elementary IP(3)-evoked puffs to elucidate whether ER [Ca(2+)] may modulate IP(3)R function via luminal regulatory sites in addition to simply determining the size of the available store and electrochemical driving force for Ca(2+) release. SERCA2b and Ca(2+) permeable nicotinic plasmalemmal channels were expressed in oocytes, and hyperpolarizing pulses were delivered to induce Ca(2+) influx and thereby load ER stores. Puffs evoked by photoreleased IP(3) were significantly potentiated in terms of numbers of responding sites, frequency and amplitude following transient Ca(2+) influx in SERCA-overexpressing cells, whereas little change was evident with SERCA overexpression alone or following Ca(2+) influx in control cells not overexpressing SERCA. Intriguingly, we observed the appearance of a new population of puffs that arose after long latencies and had prolonged durations supporting the notion of luminal regulation of IP(3)R gating kinetics. PMID- 21616534 TI - Roles of TRPM2 in oxidative stress. AB - Reactive oxygen species (ROS) play critical roles in cell death, diseases, and normal cellular processes. TRPM2 is a member of transient receptor potential (TRP) protein superfamily and forms a Ca(2+)-permeable nonselective cation channel activated by ROS, specifically by hydrogen peroxide (H(2)O(2)), and at least in part via second-messenger mechanisms. Accumulating evidence has indicated that TRPM2 mediates multiple cellular responses, after our finding that Ca(2+) influx via TRPM2 regulates H(2)O(2)-induced cell death. Recently, we have demonstrated that Ca(2+) influx through TRPM2 induces chemokine production in monocytes and macrophages, which aggravates inflammatory neutrophil infiltration in mice. However, understanding is still limited for in vivo physiological or pathophysiological significance of ROS-induced TRPM2 activation. In this review, we summarize mechanisms underlying activation of TRPM2 channels by oxidative stress and downstream biological responses, and discuss the biological importance of oxidative stress-activated TRP channels. PMID- 21616535 TI - Stimulation of human red blood cells leads to Ca2+-mediated intercellular adhesion. AB - Red blood cells (RBCs) are a major component of blood clots, which form physiologically as a response to injury or pathologically in thrombosis. The active participation of RBCs in thrombus solidification has been previously proposed but not yet experimentally proven. Holographic optical tweezers and single-cell force spectroscopy were used to study potential cell-cell adhesion between RBCs. Irreversible intercellular adhesion of RBCs could be induced by stimulation with lysophosphatidic acid (LPA), a compound known to be released by activated platelets. We identified Ca(2+) as an essential player in the signaling cascade by directly inducing Ca(2+) influx using A23187. Elevation of the internal Ca(2+) concentration leads to an intercellular adhesion of RBCs similar to that induced by LPA stimulation. Using single-cell force spectroscopy, the adhesion of the RBCs was identified to be approximately 100 pN, a value large enough to be of significance inside a blood clot or in pathological situations like the vasco-occlusive crisis in sickle cell disease patients. PMID- 21616536 TI - Bartonella species and their ectoparasites: selective host adaptation or strain selection between the vector and the mammalian host? AB - A wide range of blood-sucking arthropods have either been confirmed or are suspected as important vectors in Bartonella transmission to mammals, including humans. Overall, it appears that the diversity of Bartonella species DNA identified in ectoparasites is much broader than the species detected in their mammalian hosts, suggesting a mechanism of adaptation of Bartonella species to their host-vector ecosystem. However, these mechanisms leading to the fitness between the vectors and their hosts still need to be investigated. PMID- 21616537 TI - A Piagetian approach to infant referential behaviors. AB - Near the end of the first year of life, infants begin producing referential behaviors that in adults indicate joint attention, or coordinating shared attention to an object with another person. These behaviors have been interpreted in the social cognitive literature as an indication that infants realize they are sharing attention to an object with another person. In this paper, we address theory and research on infant referential behaviors described as joint attention and offer an alternative explanation for the presence of these behaviors. Using Piaget's constructivist theory, we show how research in this area can be interpreted without assuming that infants have advanced social cognitive abilities. We argue that infants' referential behaviors are motor signifiers of thought and that infants recognize humans as particularly relevant objects for their goal-directed behaviors. Finally, we describe how the field of infant joint attention research should proceed if a comprehensive understanding of infant cognitive processes is to be desired. PMID- 21616538 TI - Maternal sensitivity during distressing tasks: a unique predictor of attachment security. AB - The extent to which maternal sensitivity during a non-arousing free play task and during distressing tasks at 6 months predicted infant-mother attachment security was examined. When considered simultaneously, only maternal sensitivity during distressing tasks predicted subsequent attachment security. Infant temperament was unrelated to attachment security. PMID- 21616539 TI - Activation of a DeltaFOSB dependent gene expression pattern in the dorsolateral prefrontal cortex of patients with major depressive disorder. AB - BACKGROUND: A DeltaFOSB mediated transcriptional response in the nucleus accumbens (NAc) is induced by chronic social stress in rodent and a 50% down regulation of DeltaFOSB has been also reported in the NAc of eight depressed subjects. To evaluate the role of DeltaFOSB in the prefrontal cortex which is critically involved in negative cognitive bias associated with major depressive disorder (MDD) we have quantified the mRNA levels of DeltaFOSB and of five of its major target genes in the Brodmann area 46 from 24 patients with MDD (11 with psychotic symptoms) and 12 controls. METHOD: Expression of the six genes has been quantified by a real-time quantitative PCR method: DeltaFOSB, GRIA2 (encoding the GluR2 subunit of the AMPA receptor), SPARCL1 (encoding hevin), SG3 (encoding the secretogranin III), PCP4 (encoding the Purkinje cell protein 4), ATP6V0C (encoding a subunit of the lysosomal ATPase). RESULTS: Expression of DeltaFOSB and GRIA2 was significantly up-regulated (~ 1.60) in the BA 46 of MDD patients. Overexpression of SCG3 and PCP4 was restricted to psychotic subjects. The mRNA levels of GRIA2, SCG3 and PCP4 were strongly correlated in the depressed group. LIMITATIONS: All the patients were treated by antidepressants and the number of subjects in each subgroup was rather small. CONCLUSIONS: Induction of a DeltaFOSB mediated transcriptional pattern in the prefrontal cortex is opposite to the down regulation observed in the NAc. The major consequence might be a shift in the excitability of the glutamatergic synapses which depends on GluR2 (high in the NAc and low in the BA 46). PMID- 21616540 TI - Exercise and severe depression: preliminary results of an add-on study. AB - BACKGROUND: Physical exercise has been extensively researched as a therapeutic option for treatment of major depression. METHODS: In a randomized controlled trial, we analyze the effects of aerobic physical exercise as an add-on strategy for treatment of severe depressed inpatients. The exercise has a "Dose" of 16.5 kcal/kg/week, three times a week during all the hospitalization. RESULTS: Our preliminary results show that there is no significant difference in scores of Hamilton in the second week between groups (Mean[SD]=8.2[5.96] * 11.18[5.03], p=0.192). However, there is a significant reduction in Hamilton scores of patients in exercise group at discharge (Mean[SD]=5.93[4.46] * 9.45[3.56], p=0.041). Regarding Quality of Life (QoL), no significant difference were found between groups in the second week in physical domain (Mean[SD]=56.98[8.96] * 54.54[9.18], p=0.511) and psychological domain (Mean[SD]=50.88[13.88] * 42.04[12.42], p=0.106). However, there is a significant difference in psychological domain (Mean[SD]=55.88[9.92] v 41.66[13.04], p=0.004) and a trend but no statistical significance in the physical (Mean[SD]=58.80[9.14] * 52.12[8.70], p=0.07) at discharge. LIMITATIONS: Many patients receive different treatment strategies, like ECT (1 patient at exercise group * 3 at control group). Other limitation is the small number of participants included until this moment. CONCLUSION: Our preliminary results suggest that physical exercise could be a feasible and effective add-on strategy for treatment of severe depressed inpatients, improving their depressive symptoms and QoL. PMID- 21616541 TI - Excessive daytime sleepiness and fatigue in depressed patients and therapeutic response of a sedating antidepressant. AB - BACKGROUND: Although sleepiness and fatigue are common symptoms in depressed patients, the relationships among sleepiness, fatigue and treatment of depression have not been fully elucidated. The main objective of this study was to investigate the therapeutic effects of a sedating antidepressant on sleepiness and fatigue in patients with depression. METHODS: Forty-two depressed patients, who met DSM-IV diagnostic criteria, and 32 matched healthy controls participated in the baseline measurements. Sixteen of the depressed patients were treated with mirtazapine. At baseline, daytime sleepiness was assessed using the Epworth Sleepiness Scale (ESS) and Stanford Sleepiness Scale (SSS), and fatigue was assessed using the Fatigue Severity Scale (FSS) and Fatigue Impact Scale (FIS). During treatment, Multiple Sleep Latency Test (MSLT), ESS and SSS were used to measure daytime sleepiness, and the FIS, FACES Checklist-Fatigue subscale (FAF) and Fatigue Assessment Instrument (FAI) were used to measure fatigue. RESULTS: At baseline, there were significant group differences between the depressed and healthy controls on the ESS (P = 0.001), SSS (P<0.001), FSS (P < 0.001) and FIS (P < 0.001). Significant improvement of sleepiness and fatigue measures was seen after treatment with mirtazapine on the MSLT (P = 0.011), ESS (P = 0.021), SSS (P = 0.001), FIS (P = 0.002) and FAF (P = 0.004). LIMITATIONS: Open-label treatment and relatively small sample size. CONCLUSION: Daytime sleepiness and fatigue are significant symptoms in depressed patients. Slightly paradoxically, a sedating antidepressant may alleviate these symptoms. PMID- 21616542 TI - Trajectory of depression symptoms and related factors in later life--a population based study. AB - BACKGROUND: A number of studies of elderly depression have examined the association between depression and its related factors; however, they did not observe how the development of related factors influenced elderly depression over time. This study aims to understand the rising trajectory of depressive symptoms found in elderly male and female, and whether the causation of depression due to related factors would be heterogeneous between male and female over a decade from 1993 to 2003. METHOD: This study uses the data from the "Survey of Health and Living Status of the Elderly in Taiwan". A total of 1017 elderly Taiwanese, aged 60 and over, completed four survey modules. The latent growth curve model and general growth mixture model were used to identify the trajectories of depression symptoms, and to assess their related factors in the elderly male and female in Taiwan during the 10 year survey study. RESULT: The results showed that a rising trajectory of depressive symptoms came about for both Taiwan elderly male and female over ten years. We also found that in the trajectories of depressive symptoms between elderly male and female across four waves of data, the results were heterogeneous. The trajectory of depressive symptoms in males was affected by perceived health and disability; the trajectory of depressive symptoms in females was influenced by disability and social support. CONCLUSION: These findings demonstrated that the final depressive symptoms of the elderly would differ from the elderly with developmentally different depressive related factors. The findings also highlighted the developmental trajectories of depressive symptom in terms of their related factors in the elderly. PMID- 21616543 TI - Brain-derived neurotrophic factor (BDNF) plasma levels in drug-naive OCD patients are lower than those in healthy people, but are not lower than those in drug treated OCD patients. AB - BACKGROUND: Brain-derived neurotrophic factor (BDNF) plays an important role in neural development, and has been implicated in the development of depressive and anxiety disorders. Obsessive-compulsive disorder (OCD) is a chronic anxiety disorder with an unclear pathophysiology. Although genetic studies have suggested an association between BDNF and OCD, the results have been inconsistent. The aims of this study were to determine whether BDNF plasma levels in OCD patients are lower than those in healthy controls and whether BDNF plasma levels differ between drug-naive and drug-treated OCD patients. METHODS: We examined BDNF plasma levels in 22 drug-naive OCD patients, 52 drug-treated OCD patients, and 63 healthy controls. Individuals in all groups with a current or lifetime history of depression were excluded. RESULTS: BDNF plasma levels in both drug-naive OCD patients (1.97 +/- 1.80 ng/ml, p=0.00) and drug-treated OCD patients (1.98 +/- 1.54 ng/ml, p=0.00) were lower than those in normal controls (4.09 +/- 2.00 ng/ml). However BDNF plasma levels in those two OCD patients groups were not different from each other significantly (p=0.99). Length of drug treatment was positively associated with BDNF plasma levels in the drug-treated patients (r=0.34, p=0.03). LIMITATIONS: We used treatment length of two weeks and above as the criterion to recruit drug-treated patients. Probably this treatment length is not sufficient to identify drug-associated changes in BDNF levels. CONCLUSIONS: Our findings support the hypothesis that BDNF is involved in the pathophysiology of OCD, and may be a peripheral marker indicating neurotrophic impairment in OCD. PMID- 21616544 TI - Serum brain-derived neurotrophic factor and glucocorticoid receptor levels in lymphocytes as markers of antidepressant response in major depressive patients: a pilot study. AB - Depressive patients often have altered cortisol secretion, an effect that likely derives from impaired activity of the glucocorticoid receptor (GR), the main regulator of the hypothalamus-pituitary-adrenal (HPA) axis. Glucocorticoids reduce the levels of brain-derived neurotrophic factor (BDNF), a downstream target of antidepressants. Antidepressants promote the transcriptional activity of cyclic adenosine monophosphate (cAMP) response element binding protein (CREB), a regulator of BDNF expression. To identify potential biomarkers for the onset of antidepressant action in depressive patients, GR and phospho-CREB (pCREB) levels in lymphocytes and serum BDNF levels were repeatedly measured during the course of antidepressant treatment. Thirty-four depressed outpatients (10 male and 24 female) were treated with venlafaxine (75mg/day), and individuals exhibiting a 50% reduction in their baseline 17-Item Hamilton Depression Rating Scale score by the 6th week of treatment were considered responders. Responders showed an early improvement in parallel with a rise in BDNF levels during the first two weeks of treatment. Non-responders showed increased GR levels by the third week and reduced serum BDNF by the sixth week of treatment. In contrast, venlafaxine did not affect levels of pCREB. We conclude that levels of BDNF in serum and GR levels in lymphocytes may represent biomarkers that could be used to predict responses to venlafaxine treatment. PMID- 21616545 TI - Cloning and antibody recognition analysis of the canine 5-lipoxygenase gene. AB - 5-Lipoxygenase cDNA was prepared from canine white blood cells revealing the full length message using an oligonucleotide capping method. The sequenced 5 Lipoxygenase open reading frame revealed a 2031 base pair message encoding a 676 amino acid protein. The amino acid sequence showed mild variation with the presumed canine sequence, as well as differences in important residues of known phosphorylation observed in other species. The sequence had between 86 and 92% homology with other species, revealing a highly conserved sequence. Confirmation of gene product identity was achieved through transient transfection of the gene in a V5-Histidine tagged pcDNA 3.1 vector into a known canine cell line. Both V5 antibody and 5-lipoxygenase antibody confirmed the gene product using Western blotting and immunoflourescence. PMID- 21616546 TI - The importance of FMDV localisation in lymphoid tissue. AB - Foot-and-mouth disease virus, a highly contagious pathogen that can cause lameness, low weight and decreased milk production, is a scourge of agricultural livestock around the world. Although the acute phase of infection is rarely fatal, infection may persist in animals that have apparently recovered, creating a viral reservoir that some fear could contribute to the spread of disease. We have used an array of molecular techniques to search for traces of virus in tissues from the mouths and throats of infected cattle. In a carefully controlled study, we have found evidence of intact, non-replicating virus particles trapped by follicular dendritic cells within the germinal centres of lymph nodes. Strikingly, virus was present for up to 38 days post infection, even though it was undetectable in surrounding tissues. The retention of intact virus within germinal centres is likely to have a role in stimulating the long lasting immune response that is characteristic of viral infections. Our data suggests that this capture may also be responsible for preserving intact viruses capable of infecting susceptible cells as they come into contact with germinal centres. African buffalo (Syncerus caffer) are typically infected with all three South African Territories types of FMDV by 2 years of age and these viruses can be transmitted to farmed livestock. Buffalo harbour persistent virus in greater amounts and for longer periods than cattle and thus provided us with further opportunities to define the sites of viral localisation. PMID- 21616527 TI - Terutroban versus aspirin in patients with cerebral ischaemic events (PERFORM): a randomised, double-blind, parallel-group trial. AB - BACKGROUND: Patients with ischaemic stroke or transient ischaemic attack (TIA) are at high risk of recurrent stroke or other cardiovascular events. We compared the selective thromboxane-prostaglandin receptor antagonist terutroban with aspirin in the prevention of cerebral and cardiovascular ischaemic events in patients with a recent non-cardioembolic cerebral ischaemic event. METHODS: This randomised, double-blind, parallel-group trial was undertaken in 802 centres in 46 countries. Patients who had an ischaemic stroke in the previous 3 months or a TIA in the previous 8 days were randomly allocated with a central interactive response system to 30 mg per day terutroban or 100 mg per day aspirin. Patients and investigators were masked to treatment allocation. The primary efficacy endpoint was a composite of fatal or non-fatal ischaemic stroke, fatal or non fatal myocardial infarction, or other vascular death (excluding haemorrhagic death). We planned a sequential statistical analysis of non-inferiority (margin 1.05) followed by analysis of superiority. Analysis was by intention to treat. The study was stopped prematurely for futility on the basis of the recommendation of the Data Monitoring Committee. This study is registered, number ISRCTN66157730. FINDINGS: 9562 patients were assigned to terutroban (9556 analysed) and 9558 to aspirin (9544 analysed); mean follow-up was 28.3 months (SD 7.7). The primary endpoint occurred in 1091 (11%) patients receiving terutroban and 1062 (11%) receiving aspirin (hazard ratio [HR] 1.02, 95% CI 0.94-1.12). There was no evidence of a difference between terutroban and aspirin for the secondary or tertiary endpoints. We recorded some increase in minor bleedings with terutroban compared with aspirin (1147 [12%] vs 1045 [11%]; HR 1.11, 95% CI 1.02-1.21), but no significant differences in other safety endpoints. INTERPRETATION: The trial did not meet the predefined criteria for non inferiority, but showed similar rates of the primary endpoint with terutroban and aspirin, without safety advantages for terutroban. In a worldwide perspective, aspirin remains the gold standard antiplatelet drug for secondary stroke prevention in view of its efficacy, tolerance, and cost. FUNDING: Servier, France. PMID- 21616547 TI - Use of the johnin PPD interferon-gamma assay in control of bovine paratuberculosis. AB - Although the interferon-gamma (IFN-gamma) assay for measurements of cell-mediated immune (CMI) responses to paratuberculosis PPD (johnin) has been available for close to 20 years, the assay has not yet emerged as the long desired test to identify infected animals at an early time point. Among other issues, this relates to problematic interpretation of the test results and maybe an over expectation of what can be deducted from this kind of test given the chronic nature and slow development of infection of paratuberculosis. Over a number of years a modified IFN-gamma assay with addition of recombinant bovine IL-12 to the PPDj stimulation of blood samples from the heifer group in more than 20 Danish dairy herds which also perform surveillance of MAP antibodies in milk have been performed. The results indicate that IFN-gamma assay results are specific for paratuberculosis, but the IFN-gamma assay result of an individual animal cannot establish whether the animal is infected or predict the future progression of disease in this animal. The IFN-gamma assay should thus be used on a group of animals to test the level of exposure to paratuberculosis bacteria the animals have experienced, and thereby assist in maintaining rational in-herd management procedures and in the establishment of paratuberculosis status of a given herd. Indeed, for any diagnostic test applied in paratuberculosis, both the diagnostic target condition and the purpose of the diagnostic testing must be considered before any meaningful estimates of sensitivity or specificity can be given. PMID- 21616548 TI - Diversity of plasmids encoding histidine decarboxylase gene in Tetragenococcus spp. isolated from Japanese fish sauce. AB - Nineteen isolates of histamine producing halophilic bacteria were isolated from four fish sauce mashes, each mash accumulating over 1000 ppm of histamine. The complete sequences of the plasmids encoding the pyruvoyl dependent histidine decarboxylase gene (hdcA), which is harbored in histamine producing bacteria, were determined. In conjunction, the sequence regions adjacent to hdcA were analyzed to provide information regarding its genetic origin. As reference strains, Tetragenococcus halophilus H and T. muriaticus JCM10006(T) were also studied. Phenotypic and 16S rRNA gene sequence analyses identified all isolates as T. halophilus, a predominant histamine producing bacteria present during fish sauce fermentation. Genetic analyses (PCR, Southern blot, and complete plasmid sequencing) of the histamine producing isolates confirmed that all the isolates harbored approximately 21-37 kbp plasmids encoding a single copy of the hdc cluster consisting of four genes related to histamine production. Analysis of hdc clusters, including spacer regions, indicated >99% sequence similarity among the isolates. All of the plasmids sequenced encoded traA, however genes related to plasmid conjugation, namely mob genes and oriT, were not identified. Two putative mobile genetic elements, ISLP1-like and IS200-like, respectively, were identified in the up- and downstream region of the hdc cluster of all plasmids. Most of the sequences, except hdc cluster and two adjacent IS elements, were diverse among plasmids, suggesting that each histamine producers harbored a different histamine related plasmid. These results suggested that the hdc cluster was not spread by clonal dissemination depending on the specific plasmid and that the hdc cluster in tetragenococcal plasmid was likely encoded on transformable elements. PMID- 21616549 TI - Empirical Bayes estimation of random effects of a mixed-effects proportional odds Markov model for ordinal data. AB - The objective of this work was to investigate the factors influencing the quality of empirical Bayes estimates (EBEs) of individual random effects of a mixed effects Markov model for ordered categorical data. It was motivated by an attempt to develop a model-based dose adaptation tool for clinical use in colorectal cancer patients receiving capecitabine, which induces severe hand-and-foot syndrome (HFS) toxicity in more than a half of the patients. This simulation based study employed a published mixed-effects model for HFS. The quality of EBEs was assessed in terms of accuracy and precision, as well as shrinkage. Three optimization algorithms were compared: simplex, quasi-Newton and adaptive random search. The investigated factors were amount of data per patient, distribution of categories within patients, magnitude of the inter-individual variability, and values of the effect model parameters. The main factors affecting the quality of EBEs were the values of parameters governing the dose-response relationship and the within-subject distribution of categories. For the chosen HFS toxicity model, the accuracy and precision of EBEs were rather low, and therefore the feasibility of their use for individual model-based dose adaptation seemed limited. PMID- 21616550 TI - Brain metastasis as the first manifestation of small cell lung cancer in a female adolescent. AB - Small cell lung cancer is commonly regarded as an old age related disease. Although the brain is a common site of metastasis for this type of lung cancer, the first manifestation is rarely shown from the metastasis site. In this paper, we report a rare case of 19-year-old female patient with small cell lung cancer whose initial presentation was a solitary brain metastasis. The patient was admitted to our hospital for the recurrence of brain tumor, and the histological diagnosis after operation showed small cell lung cancer. Three months later, she suffered from dry cough and was diagnosed as small cell lung cancer by trans bronchial biopsy. Due to the identical cell types and immunohistochemical phenotypes, the brain tumor was retrospectively diagnosed as metastasis of small lung cancer by two neurological pathologists. She is the youngest female with brain metastasis as the first manifestation of small cell lung cancer reported so far internationally as to our knowledge. Second-hand smoking was possibly one of the risk factors. PMID- 21616551 TI - Blockage of vascular endothelial growth factor (VEGF) reduces experimental pleurodesis. AB - BACKGROUND AND OBJECTIVE: Chemical pleurodesis controls recurrent malignant pleural effusion. The mechanism that determines pleural symphysis involves the action of vascular endothelial growth factor (VEGF). We assessed the influence of the anti-VEGF antibody (bevacizumab) on pleurodesis induced by talc or silver nitrate and analyzed the temporal development of pleural angiogenesis. METHODS: Sixty New Zealand rabbits received intrapleural injection (2mL) of talc (400mg/kg) or 0.5% silver nitrate. In each group, half of the animals received an intravenous injection of bevacizumab 30min before the sclerosing agent. Five animals from each group were euthanized 7, 14, or 28 days after the procedure. Adhesions and inflammation (scores: 0-4), thickness (MUm), vascular density (vessels/field), and collagen fibers (MUm(2)) were evaluated in the visceral pleura. RESULTS: Antibody anti-VEGF interferes in pleurodesis induced by talc or silver nitrate. Pleural inflammation was discreet with no difference between the groups, regardless the anti-VEGF treatment. Concerning the vascular density of the visceral pleura, a smaller number of neoformed vessels was noted in the animals that received bevacizumab. In the animals receiving silver nitrate, the decrement in adhesions and vascular density was associated with reduced thick and thin collagen fibers, resulting in less pleural thickness. CONCLUSION: The anti VEGF antibody inhibits adhesions between pleural layers. Despite being an experimental study in animals with normal pleura, the results call attention to a likely lack of success in pleurodesis when VEGF blockers are used. PMID- 21616552 TI - Fifth Educational Symposium of the Spanish Lung Cancer Group: report on the Molecular Biology Workshop. AB - The majority of non-small-cell lung cancer (NSCLC) patients present with locally advanced (35%) or metastatic disease (40%); in this setting, it is of the utmost importance to balance efficacy with toxicity. However, with platinum combinations, survival has reached a "plateau", with median overall survival times of a mere 10-12 months, making it mandatory to search for new strategies and to identify more effective treatment. Molecular characteristics can be more informative than clinical features in predicting clinical benefit, and the identification of molecular markers can help define subgroups of patients who are likely to respond to different treatments, thus avoiding unnecessary toxicities and costs and providing the maximum benefit to each patient. Here we review research on biomarker assessment that was presented during the Molecular Biology Workshop held in Palma de Mallorca on 25 November 2010, during the Fifth Educational Symposium of the Spanish Lung Cancer Group. PMID- 21616553 TI - Decision-making under great uncertainty: environmental management in an era of global change. AB - Global change issues are complex and the consequences of decisions are often highly uncertain. The large spatial and temporal scales and stakes involved make it important to take account of present and potential consequences in decision making. Standard approaches to decision-making under uncertainty require information about the likelihood of alternative states, how states and actions combine to form outcomes and the net benefits of different outcomes. For global change issues, however, the set of potential states is often unknown, much less the probabilities, effect of actions or their net benefits. Decision theory, thresholds, scenarios and resilience thinking can expand awareness of the potential states and outcomes, as well as of the probabilities and consequences of outcomes under alternative decisions. PMID- 21616554 TI - Cellular senescence induced by cathepsin X downregulation. AB - Cellular senescence represents a powerful tumor suppressor mechanism to prevent proliferation and invasion of malignant cells. Since tumor cells as well as primary fibroblasts lacking the lysosomal cysteine-type carboxypeptidase cathepsin X exhibit a reduced invasive capacity, we hypothesized that the underlying reason may be the induction of cellular senescence. To investigate the cellular and molecular mechanisms leading to diminished migration/invasion of cathepsin X-deficient cells, we have analyzed murine embryonic fibroblasts (MEF) derived from cathepsin X-deficient mice and neonatal human dermal fibroblasts (NHDF) transfected with siRNAs targeting cathepsin X. Remarkably, both cell types exhibited a flattened and enlarged cell body, a characteristic phenotype of senescent cells. Additional evidence for accelerated senescence was obtained by detection of the common senescence marker beta-galactosidase. Further examination revealed increased expression levels of senescence-associated genes such as p16, p21, p53, and caveolin in these cells along with a reduced proliferation rate. The accelerated cellular senescence induced by cathepsin X deficiency was rescued by simultaneous expression of exogenous cathepsin X. Finally, cell cycle analysis confirmed a marked reduction of the synthesis rate and prolongation of the S phase, while susceptibility to apoptosis of cathepsin X-deficient cells remained unchanged. In conclusion, cathepsin X deficiency leads to accelerated cellular senescence and consequently to diminished cellular proliferation and migration/invasion implying a potential role of cathepsin X in bypassing cellular senescence. PMID- 21616555 TI - Differential effects of growth factors on oligodendrocyte progenitor migration. AB - Oligodendrocytes are myelinating cells of the CNS that originate as progenitor cells (OP) in discrete areas of the developing brain. During brain development, OP migrate significant distances prior to proliferating and myelinating the axons of the putative white matter tracts. Growth factors play a major regulatory role in the behavior of OP. Specifically, platelet-derived growth factor A (PDGF-A) and fibroblast growth factor 2 (FGF2) are two of the most well characterized regulators of OP development. Both growth factors interact with tyrosine kinase receptors, activating various intracellular signaling pathways. The current study advances our earlier research by comparing the effects of both PDGF-A and FGF2 on OP migration. Our results show that activation of ERK is required for OP migration. These findings correlate well with our previous demonstration of the ERK pathway mediating PDGF-A induced OP migration. We also demonstrate the significance of threshold levels of growth factors and temporal regulation for OP migration. In addition, ERK activation alone is not sufficient to induce OP migration. The current research supports the involvement of the non-ERK mediated signaling pathway in OP migration. PMID- 21616556 TI - Overexpression of osteopontin induces angiogenesis of endothelial progenitor cells via the avbeta3/PI3K/AKT/eNOS/NO signaling pathway in glioma cells. AB - Angiogenesis, a hallmark of tumor growth, is regulated by various angiogenic factors. Recent studies have shown that osteopontin (OPN) is a secreted, integrin binding protein that contributes to glioma progression. However, its effect on the angiogenesis of gliomas is not fully understood. To elucidate the role of OPN in the process of glioma angiogenesis, endothelial progenitor cells (EPCs) were treated with conditioned media of human glioma SHG44 cells overexpressing OPN. Here, we identified that OPN secreted by glioma cells accelerated EPCs angiogenesis in vitro, including proliferation, migration, and tube formation. OPN also induced the activation of AKT and endothelial nitric oxide synthase (eNOS) and increased NO production without affecting the expression of VEGF, VEGFR-1, or VEGFR-2. Moreover, the avbeta3 antibody, the PI3-K inhibitor LY294002 and the eNOS inhibitor NMA suppressed the OPN-mediated increase in NO production and angiogenesis in EPCs. Taken together, these results demonstrate that OPN directly stimulates angiogenesis via the avbeta3/PI3-K/AKT/eNOS/NO signaling pathway and may play an important role in tumorigenesis by enhancing angiogenesis in gliomas. PMID- 21616557 TI - Characterisation of a novel glycosylphosphatidylinositol-anchored mono-ADP ribosyltransferase isoform in ovary cells. AB - The mammalian mono-ADP-ribosyltransferases are a family of enzymes related to bacterial toxins that can catalyse both intracellular and extracellular mono-ADP ribosylation of target proteins involved in different cellular processes, such as cell migration, signalling and inflammation. Here, we report the molecular cloning and functional characterisation of a novel glycosylphosphatidylinositol (GPI)-anchored mono-ADP-ribosyltransferase isoform from Chinese hamster ovary (CHO) cells (cARTC2.1) that has both NAD-glycohydrolase and arginine-specific ADP ribosyltransferase activities. cARTC2.1 has the R-S-EXE active-site motif that is typical of arginine-specific ADP-ribosyltransferases, with Glu209 as the predicted catalytic amino acid. When over-expressed in CHO cells, the E209G single point mutant of cARTC2.1 cannot hydrolyse NAD(+), although it retains low arginine-specific ADP-ribosyltransferase activity. This ADP-ribosyltransferase activity was abolished only with an additional mutation in the R-S-EXE active site motif, with both of the glutamate residues of the EKE sequence of cARTC2.1 mutated to glycine (E207/209G). These glutamate-mutated proteins localise to the plasma membrane, as does wild-type cARTC2.1. Thus, the partial or total loss of enzymatic activity of cARTC2.1 that arises from these mutations does not affect its cellular localisation. Importantly, an endogenous ADP-ribosyltransferase is indeed expressed and active in a subset of CHO cells, while a similar activity cannot be detected in ovarian cancer cells. With respect to this endogenous ecto ART activity, we have identified two cell populations: ART-positive and ART negative CHO cells. The subset of ART-positive cells, which represented 5% of the total cells, is tightly maintained in the CHO cell population. PMID- 21616558 TI - Narrowband ultraviolet B phototherapy for the treatment of steroid-refractory and steroid-dependent acute graft-versus-host disease of the skin. AB - BACKGROUND: Acute graft-versus-host disease (aGvHD) is a common complication of allogeneic stem cell transplantation. It is usually treated with high doses of corticosteroids and other immunosuppressive agents. When cutaneous features are predominant, narrowband ultraviolet B (NB-UVB) phototherapy may be an attractive option for its steroid-sparing effect. OBJECTIVE: We sought to examine the clinical efficacy of NB-UVB in the treatment of steroid-refractory and steroid dependent cutaneous aGvHD. METHODS: We conducted a retrospective chart review of patients with steroid-refractory and steroid-dependent aGvHD, who received NB-UVB between 2005 and 2009 at our institution. RESULTS: We identified 14 patients with aGvHD treated with NB-UVB between 2005 and 2009. The median number of treatments was 15, administered over a median of 43 days. Eight of 14 patients (57%) achieved a complete response at the end of treatment; an additional 3 patients (21%) achieved a partial response; and 3 patients (21%) showed no improvement at the time when phototherapy was discontinued (nonresponders). Four patients developed chronic graft-versus-host disease (GvHD). Three of the 8 complete responders remained free of GvHD at 6 months' follow-up. LIMITATIONS: The rarity of steroid-refractory aGvHD limited the study to a small number of participants. Because GvHD is variable in its presentation and course, and life-threatening in many cases, large controlled prospective trials for potential therapies are difficult. CONCLUSIONS: NB-UVB is a viable option for the treatment of steroid refractory and steroid-dependent aGvHD of the skin. PMID- 21616559 TI - Segmental vitiligo associated with generalized vitiligo (mixed vitiligo): a retrospective case series of 19 patients. AB - BACKGROUND: Mixed vitiligo (MV), the association of segmental vitiligo (SV) and nonsegmental vitiligo, has been rarely reported. OBJECTIVE: The aim of this study was to delineate the clinical spectrum of MV through a case series of patients with typical SV associated with patchy bilateral vitiligo. METHODS: This was a cross-sectional evaluation in the setting of a prospective observational study conducted in the vitiligo clinic of the department of dermatology in Bordeaux, France. RESULTS: Nineteen patients with MV were identified. Four were male and 15 were female. Most patients had an onset of SV before the age of 18 years (18 of 19). In all patients, SV preceded nonsegmental vitiligo with a delay ranging from 6 months to more than 24 months. LIMITATIONS: This study was cross-sectional and based on a single-center experience. CONCLUSION: MV is not yet part of a conventional classification. However, this entity may have been neglected until now and should be included in the classification of vitiligo in addition to SV and nonsegmental vitiligo. Moreover, MV may be essential to the understanding of the pathogenesis of vitiligo as a primary skin disorder. PMID- 21616560 TI - The effects of adalimumab treatment and psoriasis severity on self-reported work productivity and activity impairment for patients with moderate to severe psoriasis. AB - BACKGROUND: Psoriasis significantly impairs work productivity and daily activities. OBJECTIVES: We sought to examine the effects of adalimumab on psoriasis-related work productivity and activity impairment and associations between the impairment and psoriasis severity in patients with moderate to severe psoriasis. METHODS: Data were from the first 16 weeks of the Randomized controlled EValuation of adalimumab Every other week dosing in moderate to severe psoriasis TriAL (REVEAL). Outcomes as measured by the Work Productivity and Activity Impairment Questionnaire for Psoriasis (WPAI-Psoriasis) included employment status, total work productivity impairment, and total activity impairment. Logistic regression and analyses of covariance were used to assess the effects of adalimumab and treatment response (>= 75% improvement in Psoriasis Area and Severity Index responders) on WPAI-Psoriasis outcomes. Longitudinal generalized estimating equations and Pearson correlation coefficients were used to assess associations between WPAI outcomes and psoriasis severity. RESULTS: Greater improvements in total work productivity impairment and total activity impairment were observed with adalimumab treatment versus placebo (15.5 and 11.1 percentage points, respectively; P < .001). Unemployment rate, total work productivity impairment, and total activity impairment were significantly associated with greater baseline psoriasis severity. Changes in WPAI outcomes were significantly correlated with greater psoriasis severity. The Dermatology Life Quality Index had stronger associations with changes in WPAI outcomes compared with clinical severity measures (Psoriasis Area and Severity Index and Physician Global Assessment). LIMITATIONS: REVEAL only included WPAI data for 16 weeks. Therefore, long-term impact of adalimumab treatment on productivity outcomes could not be assessed. In addition, information on occupational job title or industry was not collected and data were not adjusted for psoriatic arthritis. CONCLUSIONS: Adalimumab reduced psoriasis-related work productivity and activity impairment in patients with moderate to severe psoriasis. PMID- 21616561 TI - Contact sensitization in very young children. AB - BACKGROUND: Allergic contact dermatitis is an increasingly recognized clinical problem in children. OBJECTIVE: The aim of our study was to evaluate contact sensitization in patients younger than 3 years of age with suspected contact dermatitis. METHODS: During a 6-year period (2002-2008), 321 children underwent routine patch testing with a screening pediatric standard series of 30 allergens. RESULTS: Two hundred children (62.3%; 102 girls and 98 boys aged 3-36 months [mean age 27 + 5.6 months]) developed at least one positive reaction. The most frequent reactions were to nickel sulfate (26.8%), followed by potassium dichromate (9%), cocamidopropylbetaine (7.2%), cobalt chloride (6.2%), neomycin sulfate (5%), and methylchloroisothiazolinone/methylisothiazolinone (4.4%). The prevalence of contact sensitization was similar in children with (61.3%) and without (63%) atopic dermatitis. LIMITATIONS: The prevalence of contact sensitization in children younger than 3 years of age was not compared with the prevalence in older children. CONCLUSION: A high prevalence of contact sensitization was found in preschool children, even younger than 3 years of age. Patch testing should be considered in this age group when persistent dermatitis does not respond to conventional treatment. PMID- 21616562 TI - Comorbidity profiles among patients with alopecia areata: the importance of onset age, a nationwide population-based study. AB - BACKGROUND: Alopecia areata (AA) is considered an autoimmune disease with undetermined pathogenesis. Age at onset predicts distinct outcomes. A nationwide study of the relationship of AA with associated diseases stratified by onset age has rarely been reported. OBJECTIVE: We sought to clarify the role of atopic and autoimmune diseases in AA, thereby better understanding its pathogenesis. METHODS: A total of 4334 patients with AA were identified from the National Health Insurance Database in Taiwan from 1996 to 2008. A national representative cohort of 784,158 persons served as control subjects. RESULTS: Among patients with AA, there were significant associations with vitiligo, lupus erythematosus, psoriasis, atopic dermatitis, autoimmune thyroid disease, and allergic rhinitis. Different ages at onset resulted in disparate comorbidities. Increased risk of atopic dermatitis (odds ratio [OR] 3.82, 95% confidence interval 2.67-5.45) and lupus erythematosus (OR 9.76, 95% confidence interval 3.05-31.21) were found in childhood AA younger than 10 years. Additional diseases including psoriasis (OR 2.43) and rheumatoid arthritis (OR 2.57) appeared at onset age 11 to 20 years. Most atopic and autoimmune diseases were observed at onset ages of 21 to 60 years. With onset age older than 60 years, thyroid disease (OR 2.52) was highly related to AA. Moreover, patients with AA had higher risk for more coexisting diseases than control subjects. LIMITATIONS: We could not differentiate hypothyroidism from hyperthyroidism. CONCLUSIONS: AA is related to various atopic and autoimmune diseases. Different associated diseases in each onset age group of AA can allow clinician to efficiently investigate specific comorbidities. PMID- 21616563 TI - In vitro evaluation of a novel process for reducing bacterial contamination of environmental surfaces. AB - BACKGROUND: Disinfection of contaminated surfaces is an integral and challenging aspect of infection prevention. We evaluated the ability of Goldshield 5 (GS; NBS Technology, Laurelton, NY), an antimicrobial surfactant that coats surfaces with covalently bound octadecyldimethylammonium ions, to reduce the bacterial burden on contaminated surfaces. METHODS: We tested the GS product for inhibitory activity against patient isolates of methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa (PA), and Escherichia coli (EC) on fabric according to the garment industry standard American Association of Textile Chemists and Colorists 100 protocol. We also tested the product for activity against these same isolates in carrier tests with a modified Association of Official Analytical Chemists use-dilution method. RESULTS: On fabric, viability of bacterial isolates was inhibited for 14 days. GS also reduced recovery of viable MRSA, PA, and EC from Formica and stainless steel carriers treated with the product. CONCLUSION: Our results demonstrate that GS has inhibitory activity and potential utility as part of an infection control process. PMID- 21616564 TI - Device-associated infections rates in adult, pediatric, and neonatal intensive care units of hospitals in the Philippines: International Nosocomial Infection Control Consortium (INICC) findings. AB - BACKGROUND: This study investigated the rate of device-associated health care associated infection (DA-HAI), microbiological profiles, bacterial resistance, length of stay (LOS), and mortality rate in 9 intensive care units (ICUs) of 3 hospital members of the International Nosocomial Infection Control Consortium (INICC) in the Philippines. METHODS: This was an open-label, prospective cohort, active DA-HAI surveillance study of adult, pediatric, and newborn patients admitted to 9 tertiary care ICUs in the Philippines between January 2005 and December 2009, implementing methodology developed by the INICC. Data collection was performed in the participating ICUs, and data were uploaded and analyzed at the INICC headquarters using proprietary software. DA-HAI rates were registered based on definitions promulgated by the Centers for Disease Control and Prevention's National Healthcare Safety Network. RESULTS: Over a 5-year period, 4952 patients hospitalized in ICUs for a total of 40,733 days acquired 199 DA HAIs, for an overall rate of 4.9 infections per 1,000 ICU-days. Ventilator associated pneumonia posed the greatest risk (16.7 per 1,000 ventilator-days in the adult ICUs, 12.8 per 1,000 ventilator-days in the pediatric ICU, and 0.44 per 1,000 ventilator-days in the neonatal ICUs), followed by central line-associated bloodstream infections (4.6 per 1,000 catheter-days in the adult ICUs, 8.23 per 1,000 ventilator-days in the pediatric ICU, and 9.6 per 1,000 ventilator-days in the neonatal ICUs) and catheter-associated urinary tract infections (4.2 per 1,000 catheter-days in the adult ICUs and 0.0 in the pediatric ICU). CONCLUSION: DA-HAIs pose far greater threats to patient safety in Philippine ICUs than in US ICUs. The establishment of active infection control programs that involve infection surveillance and implement guidelines for prevention can improve patient safety and should become a priority. PMID- 21616565 TI - Adverse events of thiopurine immunomodulators in patients with inflammatory bowel disease. AB - BACKGROUND: Thiopurine immunomodulators are the most commonly used immunosuppressants in inflammatory bowel disease. AIMS: To evaluate the incidence of adverse events (AE) in patients with inflammatory bowel disease treated with azathioprine (AZA) or 6-mercaptopurine (MP) in our hospital, the features of these effects, the distribution of socio-demographic factors, and the possible predisposing factors. METHODS: We included 377 patients with inflammatory bowel disease who were diagnosed through 2008 and who received AZA or MP during the course of their disease. We collected retrospective demographic, clinical, and laboratory data about their disease and detailed information on any AE. RESULTS: Fifty-one patients had some form of AE with AZA or MP (13.5%) and 11% discontinued therapy because of toxicity. Statistically significant association with Crohn's disease was found (P = .008). Myelotoxicity occurred in 18 patients (4.8%) with a mean time of laboratory abnormalities appearing after 16 months. Nine patients had hepatotoxicity secondary to these drugs (2.4%); one of them developed nodular regenerative hyperplasia and portal hypertension. Ten patients had acute pancreatitis (2.7%) with a mean time occurrence of 27 days and a statistically significant association with Crohn's disease (P = .03) and smoking (P = .01). Fifteen patients had gastrointestinal intolerance (4%) but 5 were able to continue with medication given in divided doses or switching to MP. CONCLUSIONS: Thiopurine immunomodulators have a significant percentage of AE (13.5%), which, although usually mild, forced us to follow up all cases and sometimes even suspend treatment. PMID- 21616566 TI - Cytotoxic oxasqualenoids from the red alga Laurencia viridis. AB - Three new polyether squalene derivatives 15-dehydroxythyrsenol A (2), prethyrsenol A (3) and 13-hydroxyprethyrsenol A (4) have been isolated from the red alga Laurencia viridis. Their structures were determined through the interpretation of NMR spectroscopic data and chemical correlations. In addition, four semi-synthetic compounds modulating the solubility of the lead compound dehydrothyrsiferol (1) were prepared without loss of activity. The cytotoxicity of the new compounds exhibited low MUM activities. In order to explain their biological properties, docking simulations of the natural and synthetic compounds onto the alphavbeta3 integrin binding region were carried out. PMID- 21616567 TI - [Ischemic strokes in young adults and illegal drugs]. AB - One out of four ischemic strokes in France occurs in adults under 65 years old. About a third of them remain unexplained even after an extensive etiological assessment. A large part of these unexplained strokes could be linked to illegal drug abuse, and 10 % are estimated to be directly linked to illegal drugs in some international studies. The most frequently incriminated recreational drug remains cocaine, via several mechanisms. However, several other illegal drugs, some very commonly used such as cannabis, are suspected to have an important role in neurovascular diseases. In this article, we reviewed the epidemiological, pathophysiological and clinical studies, published in the international literature over the past 30 years. The drug-caused stroke epidemiology needs to be more precisely studied, as well as the underlying mechanisms depending on each drug. This is a public health issue that affects an economically active population, as stroke is the first cause of acquired handicap in adults. PMID- 21616568 TI - 'Two's company--Three's a crowd': the collection of umbilical cord blood for commercial stem cell banks in England and the midwifery profession. AB - OBJECTIVE: to explore how lay and professional stakeholders within umbilical cord blood banking discussed midwives' and privately employed phlebotomists' roles in light of commercial UCB collection, and what insights this offers of midwifery authority and power. DESIGN: qualitative study using face-to-face, semi structured interviews that were digitally recorded, transcribed and coded according to themes relating to the research aims. SETTING: across England. PARTICIPANTS: 61 interviews were conducted between April 2009 and August 2010 with lay and professional stakeholders within umbilical cord blood banking. FINDINGS: the space and access requirements for privately employed phlebotomists to conduct their work were discussed and highlighted the discursive and spatial boundary-work conducted by, or on behalf of, midwives to retain their authority over the umbilical cord blood and labour rooms. KEY CONCLUSIONS: midwives were portrayed as accommodating privately employed phlebotomists to some extent. It was implied that midwives did so because phlebotomists conformed to implicit boundaries, which required respecting midwives' authority over the labour ward, room and the umbilical cord blood. In turn, midwives' power was protected. IMPLICATIONS FOR PRACTICE: the findings highlight the important role of spatial boundaries and the significance of the organisation of spaces when implementing new services within health care. PMID- 21616569 TI - In vitro and in vivo approaches for the measurement of oral bioavailability of lead (Pb) in contaminated soils: a review. AB - We reviewed the published evidence of lead (Pb) contamination of urban soils, soil Pb risk to children through hand-to-mouth activity, reduction of soil Pb bioavailability due to soil amendments, and methods to assess bioaccessibility which correlate with bioavailability of soil Pb. Feeding tests have shown that urban soils may have much lower Pb bioavailability than previously assumed. Hence bioavailability of soil Pb is the important measure for protection of public health, not total soil Pb. Chemical extraction tests (Pb bioaccessibility) have been developed which are well correlated with the results of bioavailability tests; application of these tests can save money and time compared with feeding tests. Recent findings have revealed that fractional bioaccessibility (bioaccessible compared to total) of Pb in urban soils is only 5-10% of total soil Pb, far lower than the 60% as bioavailable as food-Pb presumed by U.S.-EPA (30% absolute bioavailability used in IEUBK model). PMID- 21616570 TI - Sources of ambient volatile organic compounds and their contributions to photochemical ozone formation at a site in the Pearl River Delta, southern China. AB - The Positive Matrix Factorization (PMF) receptor model and the Observation Based Model (OBM) were combined to analyze volatile organic compound (VOC) data collected at a suburban site (WQS) in the PRD region. The purposes are to estimate the VOC source apportionment and investigate the contributions of these sources and species of these sources to the O(3) formation in PRD. Ten VOC sources were identified. We further applied the PMF-extracted concentrations of these 10 sources into the OBM and found "solvent usage 1", "diesel vehicular emissions" and "biomass/biofuel burning" contributed most to the O(3) formation at WQS. Among these three sources, higher Relative Incremental Reactivity (RIR) weighted values of ethene, toluene and m/p-xylene indicated that they were mainly responsible for local O(3) formation in the region. Sensitivity analysis revealed that the sources of "diesel vehicular emissions", "biomass/biofuel burning" and "solvent usage 1" had low uncertainties whereas "gasoline evaporation" showed the highest uncertainty. PMID- 21616571 TI - Does small mammal prey guild affect the exposure of predators to anticoagulant rodenticides? AB - Ireland has a restricted small mammal prey guild but still includes species most likely to consume anticoagulant rodenticide (AR) baits. This may enhance secondary exposure of predators to ARs. We compared liver AR residues in foxes (Vulpes vulpes) in Northern Ireland (NI) with those in foxes from Great Britain which has a more diverse prey guild but similar agricultural use of ARs. Liver ARs were detected in 84% of NI foxes, more than in a comparable sample of foxes from Scotland and similar to that of suspected AR poisoned animals from England and Wales. High exposure in NI foxes is probably due to greater predation of commensal rodents and non-target species most likely to take AR baits, and may also partly reflect greater exposure to highly persistent brodifacoum and flocoumafen. High exposure is likely to enhance risk and Ireland may be a sentinel for potential effects on predator populations. PMID- 21616572 TI - Near, yet so far: the effect of pictorial cues on spatial attention. AB - Distinct cognitive and neural mechanisms underlie perception and action in near (within-reach) and far (outside-reach) space. Objects in far space can be brought into the brain's near-space through tool-use. We determined whether a near object can be pushed into far space by changing the pictorial context in which it occurs. Participants (n = 372) made relative length judgements for lines presented in near space, but superimposed over photographs of near and far objects. The left segment of the line was overestimated in the baseline and near context conditions whereas the right was overestimated in the far-context. The change from leftward to rightward overestimation is the same when lines are physically shifted from near to far space. Because participants did not have to do anything in relation to the photograph, the results suggest that simply viewing images with a near/far context can cause a shift of attention along the distal/proximal axis, which may reflect differential activation of the ventral/dorsal visual streams. PMID- 21616573 TI - [Coronary sinus orifice atresia]. PMID- 21616574 TI - [Effect of opening a new catheterization laboratory on myocardial infarction patients]. PMID- 21616575 TI - [Recurrent tako tsubo related to subclinical hyperthyroidism]. PMID- 21616576 TI - [Inflammation and thrombosis in atrial fibrillation]. PMID- 21616577 TI - [Sandwich stenting to treat an ostial left main narrowing following transcatheter aortic valve implantation]. PMID- 21616578 TI - [Elderly patient with platypnea-orthodeoxia: check the ascending aorta]. PMID- 21616579 TI - [Paravalvular regurgitations and percutaneous prosthetic aortic valves]. PMID- 21616580 TI - Bioremediation of Acidic and Metalliferous Drainage (AMD) through organic carbon amendment by municipal sewage and green waste. AB - Pit lakes (abandoned flooded mine pits) represent a potentially valuable water resource in hot arid regions. However, pit lake water is often characterised by low pH with high dissolved metal concentrations resulting from Acidic and Metalliferous Drainage (AMD). Addition of organic matter to pit lakes to enhance microbial sulphate reduction is a potential cost effective remediation strategy. However, cost and availability of suitable organic substrates are often limiting. Nevertheless, large quantities of sewage and green waste (organic garden waste) are often available at mine sites from nearby service towns. We treated AMD pit lake water (pH 2.4) from tropical, North Queensland, Australia, with primary treated sewage sludge, green waste, and a mixture of sewage and green waste (1:1) in a controlled microcosm experiment (4.5 L). Treatments were assessed at two different rates of organic loading of 16:1 and 32:1 pit water:organic matter by mass. Combined green waste and sewage treatment was the optimal treatment with water pH increased to 5.5 in only 145 days with decreases of dissolved metal concentrations. Results indicated that green waste was a key component in the pH increase and concomitant heavy metal removal. Water quality remediation was primarily due to microbially-mediated sulphate reduction. The net result of this process was removal of sulphate and metal solutes to sediment mainly as monosulfides. During the treatment process NH(3) and H(2)S gases were produced, albeit at below concentrations of concern. Total coliforms were abundant in all green waste-treatments, however, faecal coliforms were absent from all treatments. This study demonstrates addition of low-grade organic materials has promise for bioremediation of acidic waters and warrants further experimental investigation into feasibility at higher scales of application such as pit lakes. PMID- 21616581 TI - A review of studies applying environmental impact assessment methods on fruit production systems. AB - Although many aspects of environmental accounting methodologies in food production have already been investigated, the application of environmental indicators in the fruit sector is still rare and no consensus can be found on the preferred method. On the contrary, widely diverging approaches have been taken to several aspects of the analyses, such as data collection, handling of scaling issues, and goal and scope definition. This paper reviews studies assessing the sustainability or environmental impacts of fruit production under different conditions and identifies aspects of fruit production that are of environmental importance. Four environmental assessment methods which may be applied to assess fruit production systems are evaluated, namely Life Cycle Assessment, Ecological Footprint Analysis, Emergy Analysis and Energy Balance. In the 22 peer-reviewed journal articles and two conference articles applying one of these methods in the fruit sector that were included in this review, a total of 26 applications of environmental impact assessment methods are described. These applications differ concerning e.g. overall objective, set of environmental issues considered, definition of system boundaries and calculation algorithms. Due to the relatively high variability in study cases and approaches, it was not possible to identify any one method as being better than the others. However, remarks on methodologies and suggestions for standardisation are given and the environmental burdens of fruit systems are highlighted. PMID- 21616582 TI - Laparoscopic cryoablation versus partial nephrectomy for the treatment of small renal masses: systematic review and cumulative analysis of observational studies. AB - CONTEXT: For small renal masses (SRMs), partial nephrectomy (PN) represents the therapeutic standard of care. Laparoscopic cryoablation (LCA) could be regarded as an alternative to surgical excision in selected patients, if perioperative complication rates and oncologic results are comparable. OBJECTIVE: To perform a cumulative analysis of observational studies regarding oncologic outcomes and perioperative complications of both procedures. EVIDENCE ACQUISITION: Medline, Embase, and Web of Science searches were performed for clinically localized sporadic SRMs that were treated with PN or LCA. A total of 6785 lesions were analyzed for local and metastatic tumor progression and 10 906 procedures for perioperative complications. EVIDENCE SYNTHESIS: Patients undergoing LCA were significantly older, mean tumor sizes were lower, and mean follow-up duration was shorter (each p<0.001). Following LCA and PN, 8.5% and 1.9% developed local tumor progression, respectively (p<0.001). In multivariable analysis, the relative risk for local tumor progression of LCA versus PN was 5.24-fold increased (p<0.001); the risk of metastatic progression was similar. The overall complication rate was higher following PN (23.5% vs 17.0%; p<0.001), especially the rate of major complications (19.2% vs 10.2%; p<0.001). In multivariable analysis, the total risk for complications and major complications for PN versus LCA was 4.6-fold (p=0.004) and 9.71-fold (p<0.001) increased, respectively. Limitations of this analysis include follow-up and selection bias, and lack of standardization reporting complications and outcomes. CONCLUSIONS: Both PN and LCA are viable options for the management of SRMs. Compared with PN, LCA results in a higher risk of local tumor progression. The risk of perioperative complications appears to be lower following LCA; however, this difference is strongly influenced by selection bias, and thus limited conclusions can be made regarding true differences in complications. Therefore, PN is the gold standard for SRMs, but LCA may be indicated in selected patients with significant comorbidity. PMID- 21616583 TI - Real-time cancer cell tracking by bioluminescence in a preclinical model of human bladder cancer growth and metastasis. AB - BACKGROUND: Bladder cancer is the fifth most common malignancy in the Western world and the second most frequently diagnosed genitourinary tumor. In the majority of cases, death from bladder cancer results from metastatic disease. Understanding the multistep process of carcinogenesis and metastasis in urothelial cancers is pivotal to the development of new therapeutic strategies. Molecular imaging of cancer growth and metastasis in preclinical models provides the essential link between cell-based experiments and clinical translation. OBJECTIVE: Develop preclinical models for sensitive bladder cancer cell tracking during tumor progression and metastasis. DESIGN, SETTING, AND PARTICIPANTS: A human transitional cell carcinoma UM-UC-3 cell line was generated that stably expresses luciferase 2 (UM-UC-3luc2), a mammalian codon-optimized firefly luciferase with superior expression. Preclinical models were developed with human UM-UC-3luc2 cells xenografted into the bladder (orthotopic model with metastases) or inoculated into the left cardiac ventricle (bone metastasis model) of immunocompromised mice. MEASUREMENTS: Noninvasive, sensitive bioluminescent imaging of human firefly luciferase 2-positive bladder cancer in mice using the IVIS100 imaging system. RESULTS AND LIMITATIONS: In the orthotopic model (intravesical inoculation), tumor growth could be followed directly after inoculation of UM-UC-3luc2 cells. Importantly, micrometastatic lesions originating from orthotopically implanted cancer cells could be detected in the locoregional lymph nodes and in distant organs. In addition, the superior bioluminescent indicator firefly luciferase 2 allows the detection and monitoring of micrometastatic lesions in real time after intracardiac inoculation of human bladder cancer cells in mice. The main disadvantage is the lack of T-cell immunity in the preclinical models. CONCLUSIONS: The new bioluminescence-based preclinical bladder cancer models enable superior, noninvasive, and real-time tracking of cancer cells, tumor progression, and micrometastasis. Because of the significant improvement in detection of small cell numbers, the presented models are ideally suited for functional studies dealing with minimal residual disease as well as real-time imaging of drug response. PMID- 21616584 TI - Isolated facial palsy and ophthalmoplegia--first symptoms of a cerebral granulomatous vasculitis with giant cells--a diagnostic challenge. PMID- 21616585 TI - A case of chemotherapy-responsive paraneoplastic rubral tremor. PMID- 21616586 TI - Alpha-fetoprotein-producing pancreatic cancer cells possess cancer stem cell characteristics. AB - We aimed to demonstrate the existence of cancer stem cells in human pancreatic cancer, and to clarify that they are alpha-fetoprotein (AFP) producing cells. Six cell lines derived from human pancreatic cancers were examined, and AsPC-1 and PANC-1 were noted to express AFP. Single cell culture assays and xenotransplantation revealed that the AFP-producing cells had the capacity for self-renewal and differentiation, and that these cells were tumorigenic. Furthermore, they were resistant to anti-cancer agents. The ABCA12 transporter was expressed in the AFP-producing cells at a level more than twice as high as that in the non-AFP-producing cells. The AFP-producing cells were shown to be putative pancreatic cancer stem cells. Furthermore, the expression of ABCA12 appears to be associated with drug resistance. PMID- 21616587 TI - Reclaimed wastewater: impact on soil-plant system under tropical conditions. AB - This study investigated the ionic speciation of reclaimed urban wastewater (RWW), and the impact of increasing RWW irrigation rates on soil properties and plant nutrition under field conditions. Most RWW elements (>66%) are readily available as NH(4)(+), Ca(2+), Mg(2+), K(+), SO(4)(2-), Cl(-), H(3)BO(3), Mn(2+) and Zn(2+), but in imbalanced proportion for plant nutrition. Lead, Cd, Cr and Al in RWW are mostly bounded with DOM or OH(-).(.)Irrigation with RWW decreased soil acidity, which is beneficial to the acidic tropical soil. Although RWW irrigation builds exchangeable Na(+) up, the excessive Na(+) was leached out of the soil profile after a rainy summer season (>400 mm). Benefits of the disposal of RWW to the soil under tropical conditions were discussed, however, the over irrigation with RWW (>100% of crop evapotranspiration) led to a nutritional imbalance, accumulating S and leading to a plant deficiency of P and K. PMID- 21616588 TI - Effective heavy metal removal from aqueous systems by thiol functionalized magnetic mesoporous silica. AB - A thiol-functionalized magnetic mesoporous silica material (called SH mSi@Fe(3)O(4)), synthesized by a modified Stober method, has been investigated as a convenient and effective adsorbent for heavy metal ions. Structural characterization by powder X-ray diffraction, N(2) adsorption-desorption isotherm, Fourier transform infrared spectroscopy and elemental analyses confirms the mesoporous structure and the organic moiety content of this adsorbent. The high saturation magnetization (38.4 emu/g) make it easier and faster to be separated from water under a moderate magnetic field. Adsorption kinetics was elucidated by pseudo-second-order kinetic equation and exhibited 3-stage intraparticle diffusion mode. Adsorption isotherms of Hg and Pb fitted well with Langmuir model, exhibiting high adsorption capacity of 260 and 91.5mg of metal/g of adsorbent, respectively. The distribution coefficients of the tested metal ions between SH-mSi@Fe(3)O(4) and different natural water sources (groundwater, lake water, tap water and river water) were above the level of 10(5)mL/g. The material was very stable in different water matrices, even in strong acid and alkaline solutions. Metal-loaded SH-mSi@Fe(3)O(4) was able to regenerate in acid solution under ultrasonication. This novel SH-mSi@Fe(3)O(4) is suitable for repeated use in heavy metal removal from different water matrices. PMID- 21616589 TI - Removal of malachite green by adsorption and precipitation using aminopropyl functionalized magnesium phyllosilicate. AB - We report a method for the removal of malachite green (MG) by adsorption and precipitation using nano-sized aminopropyl functionalized magnesium phyllosilicate (AMP) clay. MG, which is used in aquaculture and fisheries, is a carcinogenic and mutagenic compound. In response to these health risks, many efforts have been focused on adsorption of MG onto various adsorbents, which is a versatile and widely used technique for removing MG from water. Herein, we describe the adsorption and precipitation of MG using AMP clay, as well as the alkaline fading phenomenon of MG. In this study, prepared AMP clay and the precipitate product after the reaction of MG-AMP clay mixture were characterized. In addition, adsorption isotherms and kinetics, as well as thermodynamic studies are presented. Based on the results, we suggest a macro- and microscopic removal mechanism for the adsorption and precipitation of MG using AMP clay. An AMP clay dosage of 0.1 mg mL(-1) exhibited a maximum removal capacity of 334.80 mg g(-1) and 81.72% MG removal efficiency. With further increases of the AMP clay dosage, removal capacity by AMP clay gradually decreased; at dosage above 0.2 mg mL(-1) of AMP clay, the removal efficiency reached 100%. PMID- 21616590 TI - Highly efficient visible light TiO2 photocatalyst prepared by sol-gel method at temperatures lower than 300 degrees C. AB - Highly efficient visible light TiO(2) photocatalyst was prepared by the sol-gel method at lower temperature (<= 300 degrees C), and characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FTIR), UV-vis diffuse reflectance spectroscopy (UV-vis DRS), X-ray photoelectron spectroscopy (XPS), Raman spectroscopy and differential scanning calorimetry-thermogravimetric analysis (DSC-TGA). The effects of the heat treatment temperature and time of the as-prepared TiO(2) on its visible light photocatalytic activity were investigated by monitoring the degradation of methyl orange solution under visible light irradiation (wavelength >= 400 nm). Results show that the as-prepared TiO(2) nanoparticles possess an anatase phase and mesoporous structure with carbon self-doping and visible photosensitive organic groups. The visible light photocatalytic activity of the as-prepared TiO(2) is greatly higher than those of the commercial TiO(2) (P-25) and other visible photocatalysts reported in literature (such as PPy/TiO(2), P3HT/TiO(2), PANI/TiO(2), N-TiO(2) and Fe(3+)-TiO(2)) and its photocatalytic stability is excellent. The reasons for improving the visible light photocatalytic activity of the as-prepared TiO(2) can be explained by carbon self-doping and a large amount of visible photosensitive groups existing in the as-prepared TiO(2). The apparent optical thickness (tau(app)), local volumetric rate of photo absorption (LVRPA) and kinetic constant (k(T)) of the photodegradation system were calculated. PMID- 21616591 TI - Degradation of tetracycline in aqueous media by ozonation in an internal loop lift reactor. AB - The degradation of tetracycline by ozone was investigated in this paper. In the laboratory scale experiments, the effect of major parameters, including pH, gas flow rate, gaseous ozone concentration, hydrogen peroxide concentration and hydroxyl radical scavenger (tert-butyl alcohol) on the degradation of tetracycline was studied. A pseudo-first order kinetic model was used to simulate the experimental results. The results indicated that the tetracycline degradation rate increased with pH, gaseous ozone concentration and gas flow rate. The addition of hydrogen peroxide or hydroxyl radical scavenger had little effect on tetracycline removal, indicating that the direct oxidation of tetracycline by ozone was dominant process and the radical contribution to the tetracycline oxidation could be neglected. The main intermediates were separated and identified as well as the simple degradation pathway of tetracycline was proposed. The COD removal reached to 35% after 90 min reaction. The acute toxicity experiments illustrated that the Daphnia magna mortality reached the maximum after 25 min ozonation and then decreased to zero after 90 min ozonation. PMID- 21616592 TI - Chemical and toxicological evolution of the antibiotic sulfamethoxazole under ozone treatment in water solution. AB - This work studied the elimination paths of the sulfonamide antibiotic sulfamethoxazole by ozonation in fast kinetic regime. The ozonation runs were performed in conditions favouring either the direct attack of the ozone molecule or the indirect attack by ozone-generated radical species with initial concentration of 0.150 mM. When doses of ozone were transferred to the liquid phase 0.2mM, in no case did sulfamethoxazole remain in solution. Two main transformation pathways were found involving the preferential attack of molecular ozone or radical pathway and leading to the formation of six intermediates, which were identified by LC-ESI-QTOF-MS. Both routes took place simultaneously in the different conditions tested, leading to a hydroxylation reaction of the benzene ring, oxidation of the amino group on the benzene ring, oxidation of the methyl group and the double bond in the isoxazole ring and S-N bond cleavage. The most abundant reaction intermediate was that resulting from S-N bond cleavage. The toxicity of partially ozonated samples for Daphnia magna and Pseudokirchneriella subcapitata revealed the formation of toxic by-products during the early stages of reaction and the persistence of considerable toxicity after the total depletion of sulfamethoxazole. PMID- 21616593 TI - Processed wastewater sludge for improvement of mechanical properties of concretes. AB - Two problems are addressed simultaneously. One is the utilisation of sludge from the treatment of wastewater. The other is the modification of the mechanical properties of concrete. The sludge was subjected to two series of treatments. In one series, coagulants were used, including ferrous sulphate, aluminium sulphate or aluminium polyhydroxychloride. In the other series, an electrochemical treatment was applied with several starting values of pH. Then, concretes consisting of a cement matrix, silica sand, marble and one of the sludges were developed. Specimens without sludge were prepared for comparison. Curing times and aggregate concentrations were varied. The compressive strength, compressive strain at yield point, and static and dynamic elastic moduli were determined. Diagrams of the compressive strength and compressive strain at the yield point as a function of time passed through the minima as a function of time for concretes containing sludge; therefore, the presence of sludge has beneficial effects on the long term properties. Some morphological changes caused by the presence of sludge are seen in scanning electron microscopy. A way of utilising sludge is thus provided together with a way to improve the compressive strain at yield point of concrete. PMID- 21616594 TI - Local characteristics of cross-unit contamination around high-rise building due to wind effect: mean concentration and infection risk assessment. AB - In this present work, the characteristics of hazardous gas dispersion and possible cross-unit contamination around a complex-shaped high-rise residential building due to wind effect are thoroughly studied using physical modeling method. Experiments were performed in a boundary layer wind tunnel for a 1:30 scale model that represented a 10-story residential building in prototype. Tracer gas, simulating exhausted room air, was continuously released from different floor levels, and its concentrations on the adjacent envelope surfaces were measured using fast flame ionization detectors. The mean concentration fields were reported and analyzed under different configurations during the experiment to consider the effects on pollutant dispersion behavior due to changes in source position and approaching wind condition, with the main emphasis on the differences between open-window and closed-window conditions. In particular, the measured concentration fields were further examined from a practical point of view, with respect to hazard assessment. Understanding these hazardous plume dispersion features is useful for employing effective intervention strategies in modern residential building environment in case of hazardous substance release. The study on this physical process is not only helpful to reduce the hazardous effect of routine release of harmful pollutant near the building, but also useful for the purpose of prevention and control of accidental infectious diseases outbreak. PMID- 21616595 TI - Novel biopolymer-coated hydroxyapatite foams for removing heavy-metals from polluted water. AB - 3D-macroporous biopolymer-coated hydroxyapatite (HA) foams have been developed as potential devices for the treatment of lead, cadmium and copper contamination of consumable waters. These foams have exhibited a fast and effective ion metal immobilization into the HA structure after an in vitro treatment mimicking a serious water contamination case. To improve HA foam stability at contaminated aqueous solutions pH, as well as its handling and shape integrity the 3D macroporous foams have been coated with biopolymers polycaprolactone (PCL) and gelatine cross-linked with glutaraldehyde (G/Glu). Metal ion immobilization tests have shown higher and fast heavy metals captured as function of hydrophilicity rate of biopolymer used. After an in vitro treatment, foam morphology integrity is guaranteed and the uptake of heavy metal ions rises up to 405 MUmol/g in the case of Pb(2+), 378 MUmol/g of Cu(2+) and 316 MUmol/g of Cd(2+). These novel materials promise a feasible advance in development of new, easy to handle and low cost water purifying methods. PMID- 21616596 TI - Efficacy of acupuncture for migraine prophylaxis: a single-blinded, double-dummy, randomized controlled trial. AB - Insufficient clinical trial data were available to prove the efficacy of acupuncture for migraine prophylaxis. A multicenter, double-dummy, single blinded, randomized controlled clinical trial was conducted at the outpatient departments of acupuncture at 5 hospitals in China to evaluate the effectiveness of acupuncture. A total of 140 patients with migraine without aura were recruited and assigned randomly to 2 different groups: the acupuncture group treated with verum acupuncture plus placebo and the control group treated with sham acupuncture plus flunarizine. Treated by acupuncture 3 times per week and drugs every night, patients from both groups were evaluated at week 0 (baseline), week 4, and week 16. The primary outcome was measured by the proportion of responders (defined as the proportion of patients with a reduction of migraine days by at least 50%). The secondary outcome measures included the number of migraine days, visual analogue scale (VAS, 0 to 10 cm) for pain, as well as the physical and mental component summary scores of the 36-item short-form health survey (SF-36). The patients in the acupuncture group had better responder rates and fewer migraine days compared with the control group (P<.05), whereas there were no significant differences between the 2 groups in VAS scores and SF-36 physical and mental component summary scores (P>.05). The results suggested that acupuncture was more effective than flunarizine in decreasing days of migraine attacks, whereas no significantly differences were found between acupuncture and flunarizine in reduction of pain intensity and improvement of the quality of life. PMID- 21616597 TI - Joint pain in people with hemophilia depends on joint status. AB - Recurrent joint bleedings in people with hemophilia (PWH) often progress into the full clinical picture of hemophilic arthropathy, accompanied by chronic pain. Although chronic pain is commonly present in PWH, investigations assessing pain thresholds have not been performed yet. Thus, the aim of this study was to obtain objective and subjective measures of joint pain in PWH and to relate these to the severity of joint pathology. Thirty-six patients (aged 43+/-11 years) with hemophilia A and B (31 severe A, 1 B; 3 moderate A, 1 B) and 40 healthy control subjects (aged 42+/-14 years) participated in this study. Mechanical pain thresholds were obtained as objective parameters using an algometer, while subjective pain intensity and quality were assessed using numeric analogue scales. Quality of life was estimated using the Short-Form Health Survey (SF-36) questionnaire. Overall, we found reduced mechanical pain thresholds as obtained from the knee (PWH--left 38.1 [28.7/57.7], right 29.5 [20.9/49.3]; control--left 67.4 [56.8/112.6], right 60.9 [42.6/97.2]), and elbow (PWH--left 23.4 [15.3/33.4], right 23.5 [20.1/35.1]; control--left 56.7 [32.6/86.6], right 53.0 [30.7/87.7] in N; median [25th/75th percentile]) joints in PWH. Interestingly, this increased pain sensitivity was related to the severity of clinical joint pathology. In addition, PWH reported their pain in a more descriptive and not affective manner and scored similar to controls in the mental domain of the SF 36, thereby indicating good coping strategies despite the chronic nature of their complaints. In conclusion, pain sensitivity at the site of the affected joints is increased and closely related to joint pathology in people with hemophilia. PMID- 21616598 TI - The influence of ethnic concordance and discordance on verbal reports and nonverbal behaviours of pain. AB - This study's aim was to examine the influence of ethnic concordance on Chinese participants' pain report and nonverbal pain expression in a laboratory setting. Participants (n=102) were exposed to a cold pressor task under 1 of 2 conditions: Chinese milieu (n=52; participants exposed to Chinese experimenters and language), or European Canadian milieu (n=50; participants exposed to Euro Canadian experimenters and English language). A reference group with 86 Euro Canadian participants, exposed to the Euro-Canadian milieu only, was included for comparison. The Chinese groups did not differ on pain intensity during the pain task. However, Chinese participants in the Chinese milieu reported significantly higher affective pain and displayed more nonverbal behaviour of pain than the Chinese participants in the Euro-Canadian milieu. In addition, compared to the Euro-Canadian group, both Chinese groups reported higher pain intensity during the pain task and greater affective pain after immersion. The results demonstrated that an ethnically concordant milieu is associated with increased nonverbal pain displays and affective pain report. These findings suggest that research on ethnic disparities in pain treatment should examine ethnic concordance between observer and individual in pain. PMID- 21616599 TI - Prevalence of Cryptosporidium spp. in pigs in Shanghai, China. AB - A survey on prevalence of Cryptosporidium spp. in pigs at 12 farms in 8 suburban districts and 1 county of Shanghai was conducted under Sheather's sucrose flotation protocol and modified acid-fast stain methods from 2006 to 2009. A total of 2323 faecal samples were collected and Cryptosporidium spp. oocysts were detected in 800 samples (34.4%). Cryptosporidium was found in all 12 pig farms. Significant variations of prevalence were observed in different farms ranging from 14.1% to 90.6%. A follow-up survey on a positive pig farm for 13 consecutive months revealed that the most serious infection of Cryptosporidium spp. in pigs happened in winter and spring, and the lowest infection season was summer. Cryptosporidium spp. infection was mainly found in piglets within 2 months and no infection was found among those pigs of 90-180 days of age. The genotype analyses were carried out through PCR-RFLP and partial sequences analysis of small subunit ribosomal RNA (SSU rRNA) in some of the positive samples. Cryptosporidium suis (57/69, 82.6%), Cryptosporidium pig genotype II (6/69, 8.7%) and mixed infection of above two species/genotype (6/69, 8.7%) were found to be the main species/genotype in pigs in Shanghai area. PMID- 21616600 TI - Mechanism of action of demethylating and immune modulatory agents--introduction. PMID- 21616601 TI - The cortisol awakening response in amyotrophic lateral sclerosis is blunted and correlates with clinical status and depressive mood. AB - Considerable evidence indicates that amyotrophic lateral sclerosis (ALS), a progressive neurodegenerative disease of the motor system, has an enormous impact on the patient's emotional and physical well-being. As previous findings indicated that particularly the rise in cortisol levels immediately after awakening, i.e., the cortisol awakening response (CAR), is associated with indices of physical and emotional well-being, we compared the CAR of 29 admitted ALS patients with that of 12 age-matched caregiver controls. Saliva samples for cortisol measurement were collected immediately, 15, 30 and 45 min after awakening. The severity of ALS progression was quantified using the ALS functional rating scale (ALSFRS) and manual muscle test (MMT). Depressive mood status in ALS patients was determined with the Beck Depression Inventory (BDI) and Hamilton Depression Rating Scale (HDRS). Salivary cortisol levels of ALS patients did not differ from those of caregiver controls at awakening, 15 min or 45 min after awakening, but were significantly lower at 30 min after awakening. Area under the curve analysis confirmed that the CAR was significantly smaller in ALS patients than in caregiver controls. A smaller CAR in ALS patients was significantly correlated to poorer clinical status, as assessed with both the ALSFRS and MMT rating instruments. Further, a smaller CAR significantly correlated with a more severe depressive mood status. No correlations were observed between total cortisol output during the first 45 min post-awakening and clinical or depressive status. In conclusion, our findings indicate that ALS patients show a blunted CAR, correlated with disease and depression severity. PMID- 21616603 TI - Cetuximab and cancers of the head and neck: tapping the circadian rhythm. AB - Proteins in tissue obtained from human skin and oral mucosa have shown a significant circadian rhythm, with the peak expression of p27 at 6:00 AM (early G1-phase marker), p53 at 10:50 AM (late G1-phase marker) and cyclin-E at 2:50 PM (S-phase marker). Patients irradiated in late afternoon/evening have shown a higher grade of mucositis and dermatitis. Studies evaluating the effect of EGFR blockade on cell cycle progression in several human cell types, including A431 squamous epithelial carcinoma cells, suggest that cetuximab leads to cell cycle arrest in G1 phase. On concurrent administration with radiation, mucositis and dermatitis are its main side-effects. So we can hypothesize that cetuximab administration after 11:00 AM would decrease these toxicities. In addition, its administration prior to late afternoon/evening (3:00 PM) can further reduce the radiation associated mucositis and dermatitis due to the occurrence of S-phase during this time and thus increase the therapeutic benefit. PMID- 21616602 TI - Chronic overexpression of corticotropin-releasing factor from the central amygdala produces HPA axis hyperactivity and behavioral anxiety associated with gene-expression changes in the hippocampus and paraventricular nucleus of the hypothalamus. AB - Environmental stress has been demonstrated to increase susceptibility for mood and anxiety disorders, and hyperactivity of the hypothalamic-pituitary-adrenal (HPA) axis, the primary endocrine response to stress, is often observed in these patients. HPA axis activation is initiated by corticotropin-releasing factor (CRF) from the hypothalamus, leading to the hypothesis that hypothalamic CRF overexpression contributes to HPA axis hyperactivity in psychiatric patients. In addition, elevated CRF in cerebrospinal fluid is observed in mood and anxiety disorder patients, suggesting that CRF is also being overproduced from extrahypothalamic sources such as the central amygdala (CeA) and overactivity of the amygdala in neuroimaging studies is a consistent finding in anxiety and depression patients. Due to the importance of CRF and the amygdala in the etiology of stress-sensitive psychiatric disorders, the present study sought to further dissect the impact of CRF overexpression (OE) in the amygdala on downstream behavioral, endocrine, and gene-expression changes typically associated with chronic stress. To test the hypothesis that elevated CRF output from the amygdala would reproduce HPA axis hyperactivity and behavioral symptoms of chronic stress, we developed a lentiviral vector in which 3.0kb of the CRF promoter drives overexpression of CRF (LVCRFp3.0CRF). In adult male rats, Experiment-1 examined behavioral consequences of chronic CRF overexpression from the amygdala; the dexamethasone (Dex)/CRF test was used to measure HPA axis reactivity. Experiment-2 focused on HPA axis disruptions; the dexamethasone suppression and CRF-stimulation tests as well as the Dex/CRF test were used. In both experiments, expression of HPA-axis related transcripts were assessed. PMID- 21616604 TI - Information homeostasis as a fundamental principle governing the cell division and death. AB - To express the genetic information with minimal error is one of the key functions of a cell. Here we propose an information theory based, phenomenological model for the expression of genetic information. Based on the model we propose the concept of 'information homeostasis' which ensures that genetic information is expressed with minimal error. We suggest that together with energy homeostasis, information homeostasis is a fundamental working principle of a biological cell. This model proposes a novel explanation of why a cell divides and why it stops to divide and, thus, provides novel insights into oncogenesis and various neuro degenerative diseases. Moreover, the model suggests a theoretical framework to understand cell division and death, beyond specific biochemical pathways. PMID- 21616605 TI - [Epidemiology of celiac disease]. AB - Celiac disease is an autoimmune disorder induced by gluten in genetically susceptible individuals. Patients may present with typical symptoms of enteropathy with diarrhoea and failure to thrive, but atypical symptoms, or even silent forms are more often recognized since serologic markers are available. This progress led to the increase of the incidence of the celiac disease during the last 30 years, which reflect probably the improvement in diagnosing atypical forms of the disease. The prevalence is comparable from a country to another, around 0,7 to 2%. There is a progression with time from latent to silent form, and then to the active disease. The exclusion of the gluten from the diet leads to the regression from the active to the latent form of the disease. PMID- 21616606 TI - Prehypertension among adults in Great Tunis region (Tunisia): A population-based study. AB - PURPOSE: The present study aimed to determine the prevalence of prehypertension (preHTN) and its cardiometabolic profile in Tunisians, and to estimate the risk for coronary heart disease (CHD) according to blood pressure status. PATIENTS AND METHOD: This cross-sectional study was conducted in 2004-2005, and used a two stage cluster sampling method to select a representative sample of the Great Tunis population. A total of 2712 individuals (1228 men and 1484 women), aged 35 to 69 years were included. Definition and classification of hypertension (HTN) was performed according to guidelines from the Joint National Committee on prevention, detection, evaluation and treatment of high blood pressure (JNC-7) report. RESULTS: The prevalence of preHTN and HTN was 56.8% and 25.0% in males, and 43.1% and 36.1% in females, respectively. Subjects with preHTN and those with HTN showed higher prevalence of diabetes, dyslipidemia, obesity and abdominal obesity than the normotensive (NT) group. The metabolic syndrome (MetS) was found in 8.0%, 17.8% and 53.8% of NT, preHT and HTN subjects, respectively. The risk of developing CHD within 10 years, as predicted by the Framingham-Anderson model, was above 15% for 3.9%, 31.1% and 65.0% among NT, preHTN and HTN subjects, respectively. In multivariate analysis, preHTN was associated with age (OR [95% CI], 1.02 [1.01-1.03]; P<0.01), male gender (2.51 [1.89-3.23]; P<0.001), obesity (2.36 [1.71-3.26]; P<0.01), abdominal obesity (1.53 [1.14-2.06]; P<0.01) and smoking (0.70 [0.53-0.92]; P<0.01). CONCLUSION: PreHTN is very common in Tunisians. It is associated with a higher prevalence of cardiometabolic risk factors and confers a higher risk for subsequent CHD. These findings support the recommendations of lifestyle modification for preHTN patients. PMID- 21616607 TI - [Immunogenetics of celiac disease]. AB - Celiac disease is an auto-immune enteropathy involving genetic factors. It is associated in almost all the patients, to specific susceptibility alleles encoding histocompatibility antigens (HLA for human leucocyte antigen), specifically certain variants of the HLA-DQ2, and the HLA-DQ8 HLA class II molecules. Its estimated prevalence is 1% in the european and north-american populations. However, although these alleles represent the main genetic factor for this disease, they do not explain it on their own, as they are expressed by up to 30% of the population. Recent immunological advances allowed identifying the immunodominant epitopes of gluten, to establish the role of tissue transglutaminase in the disease and to define at the atomic level the presentation of these antigens by the HLA-DQ molecule. It is noteworthy that the HLA susceptibility alleles only account for 40% of the whole genetic risk, and the challenge is now to explain the remaining 60%. Genome-wide association studies using the DNA arrays technology to screen single nucleotide polymorphisms to pinpoint candidate regions and genes, have started to provide answers, but contradictory results sometimes still persist. Most of the genes emerging as statistically significantly associated with celiac disease are involved in the immune response, and suggest that the situation is complex. PMID- 21616608 TI - [Symptoms of Celiac disease in childhood]. AB - The knowledge regarding celiac disease has increased dramatically in recent years, due to the availability of accurate serologic markers. Mass screening studies have shown that the prevalence of sensitization can be as high as 1/80. The range of symptoms is wide, from the classic growth failure, denutrition and diarrhea in infancy to clinically and histologically asymptomatic sensitized subjects. The interest of a routine mass screening is debated. The classical celiac disease in infancy is well known. Atypical symptoms and potentially associated disease are more frequent and potentially confounding. Physicians should be aware of any clue for celiac disease in atypical cases in order to improve the diagnostic yield, and therefore avoiding short or long term consequences. PMID- 21616609 TI - [Soft tissue infection associated with bacteremia caused by Shewanella putrefaciens]. AB - Shewanella putrefaciens is rarely involved in human infectious disease. We report here a case of soft tissue infection with bacteremia on a patient with risk factors (liver cirrhosis, diabetes mellitus). PMID- 21616610 TI - Kinetics of Equid herpesvirus type 2 infections in a group of Thoroughbred foals. AB - The significance of infection with Equid herpesvirus 2 (EHV-2) remains unresolved, mainly due to its widespread distribution, and frequent isolation of the virus not only from diseased animals, but also from clinically normal horses. It has been suggested that EHV-2 exerts its effects on the host indirectly, through predisposition to secondary infections. The aim of this study was to determine kinetics of EHV-2 infection among foals and to investigate the role that EHV-2 may play in development of Rhodococcus equi pneumonia on one farm. Serial blood samples were collected from 43 foals over a period of 10 months. Viral load of EHV-2 in blood was determined using quantitative realtime PCR assay. All but 2 foals were positive for EHV-2 on at least one occasion. The majority (88%) of foals became infected with EHV-2 within the first 2 months of life. Once infected, most foals (86%) remained positive for EHV-2 on all subsequent samplings. The load of EHV-2 varied between individual foals and between different sampling times for each foal. There was a significant difference in EHV-2 load between samples collected from healthy foals and foals suspected of R. equi pneumonia only for 2-month-old foals, but not for foals of other ages. The results of this study extend our knowledge of EHV-2 epidemiology among foals. PMID- 21616611 TI - Inter- and intra-observer agreement of Prechtl's method on the qualitative assessment of general movements in preterm, term and young infants. AB - BACKGROUND: Prechtl's method on the qualitative assessment of general movements (GMs) has been shown to be a good predictor of neurological outcome. There is substantial evidence that this method has good inter- and intra-observer agreement. AIMS: We wanted to find out whether this high agreement is reproducible in the clinical setting. STUDY DESIGN: Reliability study (inter- and intra-observer agreement). SUBJECTS: Twenty video-sequences of children at the age of preterm and writhing movements (31-41 weeks postmenstrual age) and 10 video-sequences of children at the fidgety movements age (49-54 weeks postmenstrual age) were rated by five physiotherapists. OUTCOME MEASURES: Intra- and inter-observer agreements were analyzed with percentage agreement and with nominal kappa statistic with bootstrapped bias corrected 95% confidence intervals. RESULTS: We found fair to substantial inter-observer reliability for the six response categories (time-point 1(t1): median kappa 0.44, range 0.27 to 0.59, time-point 2 (t2): median kappa 0.55, range 0.41 to 0.77) and fair to almost perfect for the normal/abnormal ratings (t1: median kappa 0.53, range 0.29 to 0.61, t2: median kappa 0.63, range 0.29 to 0.85). There was statistically significant improvement from t1 to t2 for the six response categories. The intra observer reliability for the 9-week interval was moderate to almost perfect (median kappa 0.68, range 0.41 to 0.86). CONCLUSIONS: We were not able to exactly reproduce the generally very good results. In our clinical setting now videos are evaluated by at least two trained therapists and the results are discussed, if necessary, to reach a consensus. PMID- 21616612 TI - Effectiveness of a short-term treatment with progesterone injections on synchrony of lambing and fertility in tropical hair sheep. AB - The efficacy of using a low cost system for delivering progesterone as part of an estrous synchronization protocol in sheep was evaluated. In experiment 1, Barbados Black Belly ewes (n=34) and ewe lambs (n=35; 37.5+/-0.9 kg) were assigned to be untreated, control animals (C), or to receive PGF(2alpha) on d0 (PG), or receive two injections of progesterone (200mg, i.m. each) on D -5 and on D -2.5, prior to PGF(2alpha), on D 0 (2PPG). Treatment with 2PPG increased the proportion of animals lambing to the first service (P<0.05), an effect that was greater in ewe lambs than ewes (treatment * parity; P<0.05). The interval from ram introduction to lambing and the mean lambing day was less (P=0.04) in the 2PPG-treated animals compared to control animals. In Experiment 2, lactating ewes from experiment 1 (n=61) 60-85 days postpartum were assigned within parity and number of lambs reared to remain nursing (S; n=29) or weaned (W; n=32) 3 weeks prior to treatment with the 2PPG protocol. There was no effect of treatment on the proportion of animals lambing to the first service or overall, interval from ram introduction to lambing and lambing interval. An 8-month lambing interval was observed in ewes in which estrus was synchronized regardless of physiological state. In conclusion, the two-progesterone injection synchronization protocol may be used as a practical low cost and efficient method of synchronizing estrus to reduce the lambing interval and maximize productivity in tropical breeds of sheep. PMID- 21616614 TI - Density gradient centrifugation of sperm from a subfertile stallion and effect of seminal plasma addition on fertility. AB - Stallions are not selected for fertility but for other criteria (pedigree, conformation, performances, progeny), therefore valuable but subfertile stallions with poor semen quality are frequently used in commercial breeding programs. The object of this study was to evaluate whether sperm selection through a silane coated silica colloid gradient centrifugation, with or without the addition of seminal plasma of a high fertile stallion, could improve the pregnancy rates of an oligospermic valuable stallion in a commercial breeding program. In 2008 breeding season (experiment 1, n=104 mares), simple centrifugation and density gradient centrifugation of the sperm were compared. In 2009 and 2010 breeding seasons (experiment 2, n=125 mares), the effect of the addition of 5% seminal plasma to the extender after sperm selection was evaluated. In all mares deep horn uterine insemination was performed with 1 ml containing 50*10(6) morphologically normal progressive motile spermatozoa, 24-30 h after induction of ovulation with hCG. Pregnancy diagnosis by ultrasonography was performed 14 days following ovulation. Results showed a higher per cycle pregnancy rate (P>0.05) when sperm selection through a density gradient was used (62% vs. 42.3%, exp 1), while the addition of 5% seminal plasma did not influence the outcome (45.9% vs. 47.6%, exp 2) (P>0.05). An age-related decrease in the fertility of the stallion was observed when comparing the results from the different breeding seasons (P<0.05). In conclusion, sperm selection through a discontinuous density gradient enabled a normal per cycle pregnancy rate to be achieved from an oligospermic subfertile stallion in a commercial breeding program, and no differences were observed regarding the addition of seminal plasma. PMID- 21616613 TI - Doppler ultrasonographic assessment of maternal and fetal blood flow in abnormal canine pregnancy. AB - The aim of this study was to describe the changes of uterine artery, umbilical artery and fetal abdominal aorta, renal and internal carotid arteries blood flow in abnormal canine pregnancy. Twenty-two, Brucella-negative pregnant bitches were retrospectively classified into abnormal (which had either interrupted their pregnancy between days 52 and 60 or had perinatal death >60% of the litter; n=11) and normal (which had delivered healthy puppies at term; n=11). In all the animals, color and pulsed-wave Doppler examinations of uterine artery were conducted every 10 days from Day 20 to 50 from estimated luteinizing hormone peak. Doppler ultrasonography was also conducted in the fetuses to assess umbilical artery, abdominal aorta, renal and internal carotid arteries from Day 40 to 60 of gestation. Throughout the study, resistance index (RI) of uterine, umbilical and fetal renal arteries decreased up to -15% compared to -36% (P<0.01), -11% compared to -23% (P<0.05) and 2% compared to -13% (P<0.05), respectively in the abnormal and normal bitches. Fetal abdominal aorta and internal carotid did not differ between groups (P>0.05). It is concluded that in dogs, uterine artery, umbilical artery and fetal renal artery RI differ between normal and abnormal gestation being useful for the prediction of adverse obstetric outcome. PMID- 21616615 TI - Profiling of counterfeit medicines by vibrational spectroscopy. AB - Counterfeit pharmaceutical products have become a widespread problem in the last decade. Various analytical techniques have been applied to discriminate between genuine and counterfeit products. Among these, Near-infrared (NIR) and Raman spectroscopy provided promising results. The present study offers a methodology allowing to provide more valuable information for organisations engaged in the fight against counterfeiting of medicines. A database was established by analyzing counterfeits of a particular pharmaceutical product using Near-infrared (NIR) and Raman spectroscopy. Unsupervised chemometric techniques (i.e. principal component analysis - PCA and hierarchical cluster analysis - HCA) were implemented to identify the classes within the datasets. Gas Chromatography coupled to Mass Spectrometry (GC-MS) and Fourier Transform Infrared Spectroscopy (FT-IR) were used to determine the number of different chemical profiles within the counterfeits. A comparison with the classes established by NIR and Raman spectroscopy allowed to evaluate the discriminating power provided by these techniques. Supervised classifiers (i.e. k-Nearest Neighbors, Partial Least Squares Discriminant Analysis, Probabilistic Neural Networks and Counterpropagation Artificial Neural Networks) were applied on the acquired NIR and Raman spectra and the results were compared to the ones provided by the unsupervised classifiers. The retained strategy for routine applications, founded on the classes identified by NIR and Raman spectroscopy, uses a classification algorithm based on distance measures and Receiver Operating Characteristics (ROC) curves. The model is able to compare the spectrum of a new counterfeit with that of previously analyzed products and to determine if a new specimen belongs to one of the existing classes, consequently allowing to establish a link with other counterfeits of the database. PMID- 21616616 TI - Stature estimation from long bone lengths in a Thai population. AB - The estimation of stature is a very important step in developing a biological profile for forensic identification. However, little previous work has been done on stature estimation among modern Thai people, despite a growing number of forensic cases in Thailand in recent years. The current study was carried out on a sample of 200 skeletons from a northern Thai population (132 males and 68 females), ranging in age from 19 to 94 years. The maximum lengths of six long bones (humerus, radius, ulna, femur, tibia and fibula) were measured and stature reconstruction formulae generated using linear regression. These equations were then tested on a holdout sample of 15 females and 15 males. Results reveal that the three lower limb bones are the most accurate estimators of stature among the males, with the fibula equation producing the lowest standard error of the estimate (SE=4.89cm), followed by the femur (SE=5.06cm). Results for females were mixed. The femur produced the lowest standard error among the females (SE=5.21cm), followed by the radius (SE=5.63cm). However, when tested against the holdout sample (n=30), the femur equations were considerably more accurate, with a mean absolute error of 3.5cm and a median absolute error of 2.4cm. Females exhibited a higher standard error of the estimate than reported in many previous studies. This higher error may be the result of a recent secular trend in stature affecting the females of our sample somewhat more than the males. PMID- 21616617 TI - Doxycycline reduces nitric oxide production in guinea pig inner ears. AB - OBJECTIVE: Gentamicin application is an important therapeutic option to control vertigo spells in Meniere's disease. However, even in the case of low-dose intratympanic application, gentamicin might contribute to a pathological NO increase leading to cochlear damage and hearing impairment. The study was performed to evaluate the nitric oxide (NO) reducing capacity of doxycycline in the inner ear after NO-induction by gentamicin. METHODS: In a prospective animal study, a single dose of gentamicin (10mg/kg body weight) was injected intratympanically into male guinea pigs (n=48). The auditory brainstem responses (ABRs) were recorded prior to application and 3, 5 and 7 days afterwards. The organ of Corti and the lateral wall of 42 animals were isolated after 7 days and incubated separately for 6h in cell culture medium. Doxycycline was adjusted to organ cultures of 5 animals. Two NOS inhibitors, N(G)-Nitro-l-arginine methyl ester (l-NAME) and NG-monomethyl-l-arginine monoacetate (l-NMMA), were applied in three different concentrations to the organ cultures of 30 animals in total (5 animals per concentration). As controls, seven animals received no further substance except gentamicin. The NO-production was quantified by chemiluminescence. Additional six gentamicin-treated animals were used for immunohistochemical studies. RESULTS: The ABRs declined continuously from the first to the seventh day after gentamicin application. Doxycycline reduced NO production in the lateral wall by 54% (p=.029) comparable to the effect of the applied nitric oxide inhibitors. In the organ of Corti, NO-production was reduced by about 41% showing no statistical significance in respect to great inter-animal variations. CONCLUSION: The application of doxycycline might offer a new therapeutic approach to prevent NO-induced cochlea damage through ototoxic substances. PMID- 21616618 TI - Middle ear metastasis of thymoma. AB - OBJECTIVE: We report an extremely rare case of thymoma which developed middle ear metastasis along with acute sensorineural hearing loss in the contra ear. METHOD: We present a case report and a review of the world literature concerning thymoma metastases to the middle ear. RESULTS: A 54-year-old female patient with thymoma who developed middle ear metastasis along with acute sensorineural hearing loss in the contra ear. CONCLUSION: We have not found out thymoma metastases to the middle ear in the past. PMID- 21616619 TI - Halogranum salarium sp. nov., a halophilic archaeon isolated from sea salt. AB - Three halophilic archaea, strains B-1(T), B-3 and B-4, were isolated from evaporitic salt crystals from Namhae, Korea. Cells of the strains were Gram-stain negative, motile and pleomorphic, and colonies were red-pigmented. The three isolates had identical 16S rRNA gene sequences and formed a tight phylogenetic clade with Halogranum rubrum RO2-11(T) in the genus Halogranum, showing 99.5% sequence similarity. The next most closely related species were Halogranum amylolyticum and Halogranum gelatinilyticum (97.4 and 96.3% similarity to the respective type strains). The phylogeny based on the full-length RNA polymerase subunit B' gene (rpoB') was in agreement with the 16S rRNA gene sequence analysis, but allowed better discrimination. DNA-DNA hybridization between a representative strain (B-1(T)) and the type strains of Hgn. rubrum, Hgn. amylolyticum and Hgn. gelatinilyticum revealed less than 40% relatedness. Polar lipid analysis showed that the three isolates contained phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and three glycolipids. Combined genotypic and phenotypic data supported the conclusion that strains B-1(T), B-3 and B-4 represent a novel species of the genus Halogranum, for which the name Halogranum salarium sp. nov. is proposed. The type strain is B-1(T) (=KCTC 4066(T)=DSM 23171(T)). PMID- 21616620 TI - Phase gradient imaging for positive contrast generation to superparamagnetic iron oxide nanoparticle-labeled targets in magnetic resonance imaging. AB - Positive contrast imaging methods produce enhanced signal at large magnetic field gradient in magnetic resonance imaging. Several postprocessing algorithms, such as susceptibility gradient mapping and phase gradient mapping methods, have been applied for positive contrast generation to detect the cells targeted by superparamagnetic iron oxide nanoparticles. In the phase gradient mapping methods, smoothness condition has to be satisfied to keep the phase gradient unwrapped. Moreover, there has been no discussion about the truncation artifact associated with the algorithm of differentiation that is performed in k-space by the multiplication with frequency value. In this work, phase gradient methods are discussed by considering the wrapping problem when the smoothness condition is not satisfied. A region-growing unwrapping algorithm is used in the phase gradient image to solve the problem. In order to reduce the truncation artifact, a cosine function is multiplied in the k-space to eliminate the abrupt change at the boundaries. Simulation, phantom and in vivo experimental results demonstrate that the modified phase gradient mapping methods may produce improved positive contrast effects by reducing truncation or wrapping artifacts. PMID- 21616621 TI - Renal diffusion tensor imaging: is it possible to define the tubular pathway? A case report. AB - The authors report a case of unilateral xanthogranulomatous pyelonephritis, associated with chronic lithiasis studied by standard clinical magnetic resonance imaging protocol and diffusion tensor imaging (DTI). Maps of apparent diffusion coefficient (ADC) and fractional anisotropy (FA) and tractography were reconstructed on both healthy and pathologic kidney. ADC and FA values are in agreement with the literature. Tractography reconstruction of tubular renal architecture was confirmed by histology. This result suggests the potential ability of DTI to detect structural alterations in the architecture of the kidney, as noninvasive tool, preceding the onset of clinical-laboratory alterations. PMID- 21616622 TI - An automatic cerebellum extraction method in T1-weighted brain MR images using an active contour model with a shape prior. AB - PURPOSE: The objective of this paper was to automatically segment the cerebellum from T1-weighted human brain magnetic resonance (MR) images. MATERIALS AND METHODS: The proposed method constructs a cerebellum template using five sets of 3-T MR imaging (MRI) data, which are used to determine the initial position and the shape prior of the cerebellum for the active contour model. Our formulation includes the active contour model with shape prior, which thereby maintains the shape of the template. The proposed active contour model is sequentially applied to sagittal-, coronal- and transverse-view images. To evaluate the proposed method, it is applied to BrainWeb data and a 3-T MRI data set and compared with FreeSurfer with respect to performance assessment metrics. RESULTS: The segmented cerebellum was compared with the results from FreeSurfer. Using the manually segmented cerebellum as reference, we measured the average Jaccard coefficients of the proposed method, which were 0.882 and 0.885 for the BrainWeb data and 3-T MRI data set, respectively. CONCLUSION: We presented the active contour model with shape prior for extracting the cerebellum from T1-weighted brain MR images. The proposed method yielded a robust and accurate segmentation result. PMID- 21616623 TI - Investigation of the large-scale functional brain networks modulated by acupuncture. AB - Previous neuroimaging studies have primarily focused on the neural activities involving the acute effects of acupuncture. Considering that acupuncture can induce long-lasting effects, several researchers have begun to pay attention to the sustained effects of acupuncture on the resting brain. Most of these researchers adopted functional connectivity analysis based on one or a few preselected brain regions and demonstrated various function-guided brain networks underlying the specific effect of acupuncture. Few have investigated how these brain networks interacted at the whole-brain level. In this study, we sought to investigate the functional correlations throughout the entire brain following acupuncture at acupoint ST36 (ACUP) in comparison with acupuncture at nearby nonacupoint (SHAM). We divided the whole brain into 90 regions and constructed functional brain network for each condition. Then we examined the network hubs and identified statistically significant differences in functional correlations between the two conditions. Following ACUP, but not SHAM, the limbic/paralimbic regions such as the amygdala, hippocampus and anterior cingulate gyrus emerged as network hubs. For direct comparisons, increased correlations for ACUP compared to SHAM were primarily related with the limbic/paralimbic and subcortical regions such as the insula, amygdala, anterior cingulate gyrus, and thalamus, whereas decreased correlations were mainly related with the sensory and frontal cortex. The heterogeneous modulation patterns between the two conditions may relate to the functional specific modulatory effects of acupuncture. The preliminary findings may help us to better understand the long-lasting effects of acupuncture on the entire resting brain, as well as the neurophysiological mechanisms underlying acupuncture. PMID- 21616624 TI - Detection of small intrahepatic metastases of hepatocellular carcinomas using diffusion-weighted imaging: comparison with conventional dynamic MRI. AB - PURPOSE: The purpose of our study was to compare diffusion-weighted MR imaging (DWI) with conventional dynamic MRI in terms of the assessment of small intrahepatic metastases from hepatocellular carcinoma (HCC). MATERIALS AND METHODS: In 24 patients with multifocal, small (<=2 cm) intrahepatic metastatic foci of advanced HCC, a total of 134 lesions (<=1 cm, n=81; >1 cm, n=53) were subjected to a comparative analysis of hepatic MRI including static and gadopentetate dimeglumine-enhanced dynamic imaging, and DWI using a single-shot spin-echo echo-planar MRI (b values=50, 400 and 800 s/mm(2)), by two independent reviewers. RESULTS: A larger number of the lesions were detected and diagnosed as intrahepatic metastases on DWI [Reviewer 1, 121 (90%); Reviewer 2, 117 (87%)] than on dynamic imaging [Reviewer 1, 107 (80%); Reviewer 2, 105 (78%)] (P<.05). For the 81 smaller lesions (<=1 cm), DWI was able to detect more lesions than dynamic imaging [Reviewer 1, 68 (84%) vs. 56 (69%), P=.008; Reviewer 2, 65 (80%) vs. 55 (68%), P=.031], but there was no statistically significant difference between the two image sets for larger (>1 cm) lesions. CONCLUSION: Due to its higher detection rate of subcentimeter lesions, DWI could be considered complementary to dynamic MRI in the diagnosis of intrahepatic metastases of HCCs. PMID- 21616625 TI - Assessment of clinical data of nonlinear stochastic deconvolution versus block circulant singular value decomposition for quantitative dynamic susceptibility contrast magnetic resonance imaging. AB - Dynamic susceptibility contrast magnetic resonance imaging (DSC-MRI) allows the noninvasive assessment of brain hemodynamics alterations by quantifying, via deconvolution, the cerebral blood flow (CBF) and mean transit time (MTT). Singular value decomposition (SVD) and block-circulant SVD (cSVD) are the most widely adopted deconvolution method, although they bear some limitations, including unphysiological oscillations in the residue function and bias in the presence of delay and dispersion between the tissue and the arterial input function. A nonlinear stochastic regularization (NSR) has been proposed, which performs better than SVD and cSVD on simulated data both in the presence and absence of dispersion. Moreover, NSR allows to quantify the dispersion level. Here, cSVD and NSR are compared for the first time on a group of nine patients with severe atherosclerotic unilateral stenosis of internal carotid artery before and after carotid stenting to investigate the effect of arterial dispersion. According to region of interest-based analysis, NSR characterizes the pathologic tissue more accurately than cSVD, thus improving the quality of the information provided to physicians for diagnosis. In fact, in 7 (78%) of the 9 subjects, CBF and MTT maps provided by NSR allow to correctly identify the pathologic hemisphere to the physician. Moreover, by emphasizing the difference between pathologic and healthy tissues, NSR may be successfully used to monitor the subject's recovery after the treatment and/or surgery. NSR also generates dispersion level and non-dispersed CBF and MTT maps. The dispersion level provides information on CBF and MTT estimates reliability and may also be used as a clinical indicator of pathological tissue state complementary to CBF and MTT, thus increasing the clinical information provided by DSC-MRI analysis. PMID- 21616626 TI - Evaluation of the web-based Diabetes Interactive Education Programme (DIEP) for patients with type 2 diabetes. AB - OBJECTIVE: The objective in this study was to evaluate a web-based type 2 diabetes self-management education programme aimed at improving knowledge, encouraging active patient participation and providing supportive self-management tools. METHODS: (1) An effect evaluation was conducted using a randomized controlled trial with a pre-test and post-test design (n=99) and a knowledge questionnaire. (2) A user evaluation was conducted using an online questionnaire (n=564) and one-on-one interviews (n=11) to examine the perceived quality, use of functionalities and use of the programme as a supportive tool in education. RESULTS: The effect evaluation showed a significant intervention effect (p<0.01) on knowledge. The user evaluation showed high satisfaction with the programme's content, credibility and user-friendliness. However, functionalities and self management tools were used by less than half of the participants. CONCLUSION: The programme can improve knowledge, but it is not fully used as intended. A more optimal use of the programme is necessary for higher efficacy. PRACTICE IMPLICATIONS: The use of mostly spoken text instead of written text was highly appreciated and could be used more often for educational websites. Furthermore, health care practitioners need support in implementing new educational programmes during consultations. PMID- 21616627 TI - Diabetes Medication Assistance Service: the pharmacist's role in supporting patient self-management of type 2 diabetes (T2DM) in Australia. AB - OBJECTIVE: To evaluate the capacity and effectiveness of trained community pharmacists in delivering the Diabetes Medication Assistance Service (DMAS) via (1) number and types of self-management support interventions (SMSIs); (2) number of goals set and attained by patients and (3) patient outcomes (glycaemic control, medication adherence and satisfaction). METHODS: Pharmacists (n=109) from 90 community pharmacies in Australia were trained and credentialed to deliver the DMAS. The training focused on developing pharmacists' knowledge and skills in supporting patients' diabetes self-management. RESULTS: A total of 387 patients completed the trial. The mean number of SMSIs per patient was 35 (SD +/ 31) and the majority (87%) had at least one documented goal that was fully or partially attained. There were significant health benefits for patients including improved glycaemic control and a reduced risk of non-adherence to medications. Over 90% of DMAS patients reported improvements in their knowledge about diabetes self-management. CONCLUSION: The DMAS provides self management support in the community pharmacy for people with T2DM which may result in improved clinical outcomes. PRACTICE IMPLICATION: Given appropriate training in diabetes care and behavior change strategies, community pharmacists can offer programs which provide self-management support to their patients with T2DM and improve their health outcomes. PMID- 21616628 TI - Subcutaneous emphysema of the upper extremity after elbow arthroscopy. AB - Subcutaneous emphysema of the upper extremity is rare. Crepitation on physical examination and visible gas on radiographs raise the concern of gas gangrene due to gas-producing bacteria. Rapid establishment of a differential diagnosis is necessary to initiate proper treatment. We present a case of subcutaneous emphysema after elbow arthroscopy caused by a noninfectious genesis. A 59-year old woman with loose bodies in her left elbow due to mild degenerative joint disease and restricted range of motion was offered an elbow arthroscopy with removal of loose bodies and arthrolysis. Postoperatively, the elbow was actively put alternatively in maximum extension and flexion. On the first postoperative day, rapidly ascending swelling and subcutaneous crepitation starting from the hand to the forearm were noted. There was no clinical evidence of infection. Radiographs showed subcutaneous air. Frequent blood tests and clinical evaluation ruled out a potentially life-threatening bacterial infection, and the signs resolved after 1 week without surgical treatment. Presumably, the intensive postoperative range-of-motion exercises led to a sucking in of air into the wound during each movement. This case illustrates that it is important to differentiate nonbacterial from bacterial causes of soft-tissue gas formation to initiate the appropriate treatment. PMID- 21616629 TI - [Deployment of the checklist "Patient safety in the operating room" in two Lorraine hospitals. Performances and difficulties]. AB - OBJECTIVE: To evaluate the implementation of the checklist HAS 2010 in two Lorraine health facilities to identify objectively brakes and levers to suggest possible improvements. STUDY DESIGN: Descriptive survey. MATERIAL AND METHODS: Operating rooms of the Neurosciences building Central University Hospital, Nancy, Belle-Isle Hospital participating public hospital service, Metz. Two stages: a retrospective audit allowed to objectify quantitatively the presence of CL in the patient record and monitor compliance with the instructions of filling. A survey by self-administered questionnaire to professionals to assess the use of CL. RESULTS: CL was present in 50 % and 100 % records. The filling did not comply with instructions of the HAS (occupancy, three different times, time out before incision, stop the procedure if "no") and it was hard for about 30 % of participants, some items have problems of understanding. CL has strengthened information sharing about one third of respondents, over half of them are of interest, it has already helped to prevent errors. The deployment strategy influences directly the involvement of professionals. The evaluation of the use of CL cannot be done only through an audit record; the association with a self administered questionnaire appears to be relevant. CONCLUSION: Good adhesion to the concept, hope for better results after stock enhancement: optimize information and training of all professionals, avoid duplication with other documents. The audit record alone is not a good tool for evaluating the use of LBC, we must associate a self-administered questionnaire. PMID- 21616630 TI - Enoyl coenzyme A hydratase 1 is an important factor in the lymphatic metastasis of tumors. AB - We have previously demonstrated that enoyl coenzyme A hydratase 1 (Ech1) is involved in the lymphatic metastasis of tumors. In this study, RNAi was used to investigate the role of Ech1 in Hca-F, a hepatocarcinoma cell line with high rates of lymphatic metastasis. The downregulation of Ech1 inhibited proliferation of the Hca-F cells, increased the ratio of Hca-F cells in S phase to G(1) phase and decreased the adhesion and migration capacities of Hca-F cells. A higher expression level of Ech1 was confirmed in tissue from patients with gastric carcinoma (GC) with lymph node metastases (LNM), indicating the clinical association with tumor metastasis. The results indicate that Ech1 is a critical factor in the development of lymphatic metastasis in these tumors. PMID- 21616631 TI - Pyrrolo[2,3-alpha]carbazole derivatives as topoisomerase I inhibitors that affect viability of glioma and endothelial cells in vitro and angiogenesis in vivo. AB - Topoisomerase I is one of the most significant molecular targets through which indolocarbazoles inhibit tumour growth. In the present work, we studied the effect of new pyrrolo[2,3-alpha]carbazole derivatives on topoisomerase I activity in vitro, as well as on the viability of glioma and endothelial cells in vitro and on angiogenesis in vivo. All the tested compounds significantly decreased topoisomerase I activity in a concentration dependent manner, with the most effective being 1c, 1d(1), 1d(2) and 1f. The number of viable glioma and endothelial cells in vitro was also decreased in a concentration-dependent manner by all the tested compounds, although efficacy and potency differed in endothelial compared with glioma cells. Compounds 1c, 1d(1), 1e and 1f were the most effective in glioma cells, while compounds 1d(2) and 1e were the most effective in decreasing the number of viable endothelial cells. Finally, all the tested compounds inhibited angiogenesis in the chicken embryo chorioallantoic membrane in a significant and dose-dependent manner, with the most effective inhibitor being compound 1d(2). These data suggest that the tested pyrrolo[2,3 alpha]carbazole derivatives inhibit topoisomerase I activity and may be potentially useful for inhibition of glioma cell growth and angiogenesis. PMID- 21616632 TI - P21 (waf1/cip1) is required for non-small cell lung cancer sensitive to Gefitinib treatment. AB - Lung cancer is the leading cause of death from cancer in the world. Gefitinib is known to its inhibition of EGFR tyrosine kinase and worldwide used for antitumor in non-small cell lung cancer (NSCLC). Here, we show that Gefitinib reduces p-Akt levels, concomitant with elevation of p21 levels and suppression of cdk2/4 and cyclinE/D1 activities which result in impaired cell cycle progression through G1 arrest only in NSCLC cells in which it inhibits growth. We find that Gefitinib induced p21 protein stability, rather than increased RNA accumulation, was responsible for the elevated p21 levels. More, treatment of beta-elemene, a natural plant drug extracted from Curcuma wenyujin, restored sensitivity to Gefitinib via the mechanism modulated the elevation of p21 levels in the cells which are acquired resistance to Gefitinib. These data suggest that administration of Gefitinib in combination with beta-elemene may offer great opportunities for NSCLC which are acquired resistance to Gefitinib. The p21 effect on the cells to response to Gefitinib was further confirmed by p21 over expression and knockdown studies pointing to a requirement of p21 for the cells sensitive to Gefitinib. Thus, we propose that p21 is required for Gefitinib sensitive NSCLC cells. PMID- 21616633 TI - Protective effects of tryptophan on neuro-inflammation in rats after administering lipopolysaccharide. AB - Tryptophan (TRP), which plays an important role in immune system regulation, protein synthesis, serotonin (5-HT) and melatonin production, is a potent endogenous free radical scavenger and antioxidant. The aim of this work was to determine the efficacy of TRP in neuro-inflammation induced by systemic administration of lipopolysacharide (LPS, 20mg/kg) which promotes the synthesis of free radical (LPO: MDA and 4-HDA), and pro-inflammatory cytokine Interferon gamma (IFN-gamma) in different brain regions (cerebral cortex and hippocampus) of rats. Experiments were performed on adult female, pregnant and lactating rats fed with a diet of TRP content (0.5mg/100g protein), cerebral cortex and hippocampus were evaluated for lipid peroxidation (LPO) products, nitrites, nitrates and plasmatic concentration of IFN-gamma. LPO levels in LPS+TRP groups were significantly decreased than that obtained in the LPS group. However, there were no observed differences in plasmatic levels of nitrites and nitrates as well as IFN-gamma, neither in the cerebral cortex or hippocampus. The TRP has protective effect in the oxidative damage in a model of endotoxic shock in the breading nurslings induced by the systemic administration of LPS, acting as a scavenger of free radicals. So, it can be proposed as an innocuous protector agent in the endotoxic shock process. PMID- 21616634 TI - The ideal patient advocate. PMID- 21616635 TI - Fibromuscular dysplasia associated with simultaneous spontaneous dissection of four peripheral arteries in a 30-year-old man. AB - A 30-year-old man had a sudden bout of severe abdominal pain. An enhanced computed tomographic scan revealed dissections of the celiac artery, superior mesenteric artery, left renal artery, and right external iliac artery; stenosis of the right renal artery; and left kidney infarction. After careful evaluation, the patient was diagnosed with fibromuscular dysplasia (medial dysplasia), based on the findings obtained from the enhanced computed tomographic scan. This case is extremely rare because fibromuscular dysplasia occurred concurrently with simultaneous spontaneous dissections of four peripheral arteries in a young man. PMID- 21616636 TI - Carotid endarterectomy performed after carotid artery restenosis following stent migration. AB - Right carotid occlusion and left carotid stenosis were determined in a patient who had undergone a cerebrovascular accident. We performed a carotid endarterectomy for the high-risk patient who developed middle cerebral artery occlusion during stenting and a restenosis owing to stent migration. PMID- 21616637 TI - Multiscale interactions between chemical and electric signaling in LTP induction, LTP reversal and dendritic excitability. AB - Synaptic plasticity leads to long-term changes in excitability, whereas cellular homeostasis maintains excitability. Both these processes involve interactions between molecular events, electrical events, and network activity. Here I explore these intersections with a multilevel model that embeds molecular events following synaptic calcium influx into a multicompartmental electrical model of a CA1 hippocampal neuron. I model synaptic plasticity using a two-state (bistable) molecular switch that controls glutamate receptor insertion into the post synaptic density. I also model dendritic activation of the MAPK signaling pathway, which in turn phosphorylates and inactivates A-type potassium channels. I find that LTP-inducing stimuli turn on individual spines and raise dendritic excitability. This increases the amount of calcium that enters due to synaptic input triggered by network activity. As a result, LTD is now induced in some synapses. Overall, this suggests a mechanism for cellular homeostasis where strengthening of some synapses eventually balances out through weakening of a possibly overlapping set of other synapses. Even in this very narrow slice of cellular events, interesting system properties arise at the interface between multiple scales of cellular function. PMID- 21616638 TI - The clinical relevance of orthognathic surgery on quality of life. AB - The aim of orthognathic surgery is to produce a more aesthetic facial skeletal appearance, and improve jaw function. This prospective study, aimed to evaluate the impact of orthognathic surgery on quality of life for patients with dentofacial deformity, and whether it was clinically meaningful. 62 consecutive patients were recruited (27 male, 35 female) aged 18-38 years. Baseline data were collected using a validated health status measure (Orthognathic Quality of Life Questionnaire (OQLQ)) and a visual analogue scale (VAS). Postoperative questionnaires (OQLQ, VAS) and a Global Transition Scale (GTS) were completed at 6 months after completion of treatment and compared with pre-treatment scores. Following surgery, there was a significant (p<0.05, paired t test) improvement in OQLQ scores for each domain. The proportion of patients reporting a moderate or large improvement was: facial appearance (93%), chewing function (64%), comfort (60%) and speech (32%). Clinical relevance of change scores was reported in terms of effect sizes, and the largest effect was on facial aesthetics. The clinical impact was moderate on social aspects of deformity and oral function and a small effect on awareness of facial deformity. This research reaffirms that orthognathic surgery has positive effects on quality of life. PMID- 21616639 TI - Using Ferguson mouth gag to improve the visibility and surgical access in TMJ surgery (Arakeri's TMJ Technique). PMID- 21616640 TI - Pentraxin 3 as a biomarker for acute coronary syndrome: comparison with biomarkers for cardiac damage. AB - BACKGROUND: Pentraxin 3 (PTX3) is increased in circulating blood during the acute stage of acute coronary syndrome (ACS). Therefore, we compared diagnostic values of PTX3 for ACS with those of biomarkers for myocardial damage, such as troponin T (TnT) and heart-type fatty acid binding protein (H-FABP). METHODS AND RESULTS: Patients (n = 87) undergoing coronary angiography (CAG), consisting of 16 ACS and 71 non-ACS patients were enrolled. Non-ACS consists of 12 patients with normal CAG, 30 stable angina pectoris (SAP) patients controlled by medical treatment, and 29 SAP patients who required elective coronary revascularization (percutaneous coronary intervention or coronary artery bypass graft). Age, gender, or prevalence of diabetes, hypertension, or smoking was not significantly different between ACS and non-ACS groups. Serum total, high-density lipoprotein, or low-density lipoprotein cholesterol, or triglyceride levels were not significantly different between ACS and non-ACS. PTX3 levels were not significantly correlated with lipid profiles or different between those with and without conventional risk factors. Circulating PTX3, TnT, and H-FABP levels were significantly higher in ACS than non-ACS. In receiver-operating characteristic (ROC) curves, area under the curve (AUC) values for PTX3, TnT and H-FABP were 0.920, 0.674, and 0.690, respectively. ROC curves of PTX3 (AUC: 0.901), TnT (AUC: 0.731), and H-FABP (AUC: 0.633) for ST-elevation ACS were similar to those for whole ACS. In a TnT-negative subgroup, the AUC values of PTX3 and H-FABP for ACS were 0.981 and 0.489, respectively. CONCLUSIONS: PTX3 is a sensitive and specific biomarker for the diagnosis of ACS, and shows additional diagnostic values when measured in combination with TnT. PMID- 21616641 TI - Patients with more coronary yellow plaques have higher risk of stenosis progression within 7 months. AB - BACKGROUND: Disruption of vulnerable plaques causes acute coronary syndrome and stenosis progression. Yellow plaques are regarded as vulnerable and the number of yellow plaques per vessel (NYP) has been reported as a marker of vulnerable patients. Therefore, we examined if patients with more yellow plaques would have higher risk of stenosis progression. METHODS AND RESULTS: A series of patients (n = 70) who received percutaneous coronary intervention (PCI) and angioscopy was included. Patients were divided into 2 groups according to NYP: group 1 (NYP <4, n = 32) and group 2 (NYP >= 4, n = 38). Coronary artery stenosis progression in any segment excluding target lesion of PCI was examined by angiography at 7 months. Maximum yellow color grade of yellow plaques (2.7 +/- 0.7 vs. 1.7 +/- 1.2, p < 0.0001) and the number of non-target disrupted yellow plaques was larger in group 2 than in group 1 (1.1 +/- 1.5 vs. 0.2 +/- 0.6, p=0.0017). Progression of coronary stenosis was detected more frequently in group 2 than in group 1 (29% vs. 9%, p = 0.041). The number of sites with stenosis progression was larger in group 2 than in group 1 (0.47 +/- 0.98 vs. 0.09 +/- 0.30 sites/patient, p = 0.036). CONCLUSIONS: Vulnerable patients with more yellow plaques had higher incidence of stenosis progression. Approximately 30% of vulnerable patients with NYP >= 4 had stenosis progression within 7 months. PMID- 21616642 TI - Restenosis after implantation of sirolimus-eluting stent begins suddenly, shows short term progression, and stops suddenly. AB - BACKGROUND AND PURPOSE: The peak of restenosis in patients implanted with bare metal stents (BMS) is thought to be 6 months after BMS implantation, but the development of restenosis with respect to time and the peak of restenosis in patients implanted with drug-eluting stents (DES) is not known. This study aims to reveal the rate of development of restenosis with respect to time in patients implanted with DES. METHODS: A total of 282 patients who underwent sirolimus eluting stent (SES) implantation in native coronary arteries at our hospital were evaluated by serial quantitative angiography at 3 and 6 months, and based on the latter results, at 1 and 2 years after SES implantation. Clinical data were collected for up to 3 years. RESULTS: Three-year follow-up data were obtained for 261 patients. The 3-year incidence of clinically driven target-lesion revascularization (TLR) was 6.1% (16/261); of the 16 cases, 5 occurred at 3-month follow-up, 7 at 6-month angiographic follow-up, and 1 at 1-year follow up, respectively. While minimum lumen diameter (MLD) of these vessels that underwent TLR at 6 months decreased rapidly after the 3-month angiographic follow-up, MLD of the vessels with 50-70% stenosis at 6-month angiographic follow-up was almost unchanged at 1-year angiographic follow-up; however, 3 lesions required late (i.e. beyond 1 year) revascularization. CONCLUSIONS: It is difficult to predict SES restenosis by angiography. SES restenosis begins suddenly, shows short-term progression, and stops suddenly. However, treatment of de novo coronary stenosis with SES is associated with a sustained clinical benefit and a very low incidence of TLR. PMID- 21616643 TI - Epileptic EEG classification based on extreme learning machine and nonlinear features. AB - The automatic detection and classification of epileptic EEG are significant in the evaluation of patients with epilepsy. This paper presents a new EEG classification approach based on the extreme learning machine (ELM) and nonlinear dynamical features. The theory of nonlinear dynamics has been a powerful tool for understanding brain electrical activities. Nonlinear features extracted from EEG signals such as approximate entropy (ApEn), Hurst exponent and scaling exponent obtained with detrended fluctuation analysis (DFA) are employed to characterize interictal and ictal EEGs. The statistics indicate that the differences of those nonlinear features between interictal and ictal EEGs are statistically significant. The ELM algorithm is employed to train a single hidden layer feedforward neural network (SLFN) with EEG nonlinear features. The experiments demonstrate that compared with the backpropagation (BP) algorithm and support vector machine (SVM), the performance of the ELM is better in terms of training time and classification accuracy which achieves a satisfying recognition accuracy of 96.5% for interictal and ictal EEG signals. PMID- 21616644 TI - Molecular detection of OXA carbapenemase genes in multidrug-resistant Acinetobacter baumannii isolates from Iraq and Georgia. AB - The aim of this study was to determine the susceptibility to imipenem (IPM) of Acinetobacter baumannii isolates from different countries and to characterise the carbapenemase-encoding genes in IPM-resistant isolates. A total of 12 A. baumannii strains collected in Belgium (n=2), Iraq (n=8) and Georgia (n=2) were included in the study. Identification of the isolates was confirmed using matrix assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS). Antibiotic susceptibility testing was performed by the disk diffusion method, and Etest was used to determine the IPM minimum inhibitory concentrations (MICs) of resistant isolates. The presence of carbapenemase-encoding genes was investigated by polymerase chain reaction (PCR). All A. baumannii isolates were eventually identified by MALDI-TOF MS with high score values. Amongst the 12 strains, 6 were found to be resistant to IPM (MICs >=16 MUg/mL), comprising clinical isolates from wound infections of soldiers who were injured either during the Iraq war in 2007 (5 isolates) or during the Georgian-Russian war in 2008 (1 isolate from Georgia). All isolates contained ISAba1 and bla(OXA-51 like), but isolates from Iraq contained the bla(OXA-23) gene located on a plasmid whereas the isolate from Georgia contained the bla(OXA-24) gene located on the chromosome. None of the IPM-resistant isolates contained the bla(OXA-58)- or bla(NDM-1)-encoding genes. In conclusion, these results re-emphasise the worldwide dissemination of OXA carbapenemase genes in multidrug-resistant clinical isolates of A. baumannii and, to the best of our knowledge, report the first IPM-resistant A. baumannii strain isolated from a patient during the Georgian-Russian war with the bla(OXA-24) gene located on the chromosome. PMID- 21616646 TI - Prevalence and impact of cannabis use disorders in adolescents with early onset first episode psychosis. AB - BACKGROUND: Previous studies on the impact of cannabis use disorders (CU) on outcome in psychosis were predominantly based on non representative samples, often have not controlled for confounders and rarely focused on adolescent patients. Thus, the aims of the present study were to assess: (i) prevalence of CU; (ii) baseline and pretreatment differences between CU and those without CU (NCU); (iii) the impact of baseline and course of CU on 18-month outcomes in a representative cohort of adolescents with early onset first episode psychosis (EOP). METHODS: The sample comprised 99 adolescents (age 14 to 18) with EOP (onset age 14 to 17), admitted to the Early Psychosis Prevention and Intervention Centre in Australia. Data were collected from medical files using a standardized questionnaire. RESULTS: Prevalence of lifetime CU was 65.7%, of current CU at baseline 53.5%, and of persistent CU throughout treatment 26.3%. Baseline CU compared to NCU had significantly higher illness-severity, lower psychosocial functioning, less insight, lower premorbid functioning and longer duration of untreated psychosis. Compared to all other groups, only persistent CU was linked to worse outcomes and more service disengagement. Effect sizes were medium controlling for relevant confounders. Medication non-adherence did not explain the association between persistent CU and worse outcome. CONCLUSIONS: Baseline CU was associated with worse baseline characteristics, but only persistent CU was linked with worse outcome. About half of those with baseline CU reduced cannabis during treatment. For these, effectively treating the psychotic disorder may already be beneficial. However, future research is necessary on the reasons for persistent CU in EOP and its treatment. PMID- 21616645 TI - No gender differences in social outcome in patients suffering from schizophrenia. AB - Differences between female and male patients with schizophrenia in psychopathology and course of illness have frequently been reported. However, the influence of sex on symptomatic and social remission is still an open issue. In the present study, differences between males and females in both clinical and social remission rates and in scores on several scales assessing social functioning were evaluated in 295 stabilized patients with schizophrenia, schizoaffective or delusional disorder. Female patients, as compared with males, showed a later onset of the illness, less negative symptoms and less frequent alcohol abuse. No significant difference was found between females and males in the rate of symptomatic and functional remission. No significant effect of sex was observed on any index of social functioning. PMID- 21616647 TI - Biosynthesis and characterization of gold nanoparticles produced by laccase from Paraconiothyrium variabile. AB - During recent years investigation on the development of eco-friendly processes for production of gold nanoparticles (GNPs) have received much attention due to hazardous effects of chemical compounds used for nanoparticle preparation. In the present study, the purified laccase from Paraconiothyrium variabile was applied for synthesis of Au nanoparticles (AuNPs) and the properties of produced nanoparticles were characterized. The UV-vis spectrum of formed AuNPs showed a peak at 530 nm related to surface plasmon absorbance of GNPs represented the formation of gold nanoparticles after 20 min incubation of HAuCl(4) (0.6 mM) in the presence of 73 U laccase at 70 degrees C. Transmission electron microscopy (TEM) image of AuNPs showed well dispersed nanoparticles in the range of 71-266 nm as determined by the laser light scattering method. The pattern of energy dispersive X-ray (EDX) of the prepared GNPs confirmed the structure of gold nanocrystals. PMID- 21616648 TI - Synthesis and aggregation behaviour of a new sultaine surfactant. AB - The aggregation properties of a new sultaine surfactant have been studied in buffered aqueous solution at pH 7.4 under controlled condition of osmolarity. Spontaneously formed sultaine vesicles with a mean diameter of about 1 MUm can be observed by optical microscopy. The phase behaviour of the surfactant has been investigated by differential scanning calorimetry (DSC) and Nile Red fluorescence. Two critical vesicular concentrations (CVC(1) and CVC(2)) have been fluorimetrically measured, by using pyrene and Nile Red as the fluorescent probes. The two populations of vesicles behave differently as a consequence of their size. The stability of extruded large unilamellar vesicles (LUV) formed slightly above the CVC(1) has been evaluated in the temperature range 25-75 degrees C by following the rate of spontaneous release of entrapped 5(6) carboxyfluorescein (CF). The stability of the same vesicles at 70 degrees C has also been investigated under osmotic stress obtained by adding NaCl or sucrose to the bulk solution. At a sultaine concentration above the CVC(2) LUV tend to associate and form stable larger closely packed aggregates as suggested by Dynamic Laser Light Scattering and rheological measurements. PMID- 21616649 TI - Carbon nanotube-chitosan modified disposable pencil graphite electrode for vitamin B12 analysis. AB - A single walled carbon nanotube-chitosan (SWCNT-chitosan) modified disposable pencil graphite electrode (PGE) was used in this study for the electrochemical detection of Vitamin B(12). Electrochemical behaviors of SWCNT-chitosan PGE and chitosan modified PGE were compared by using cyclic voltammetry (CV), square-wave voltammetry (SWV) and electrochemical impedance spectroscopy (EIS) techniques. SWCNT-chitosan modified electrode was also used for the quantification of Vitamin B(12) in pharmaceutical products. The results show that this electrode system is suitable for sensitive Vitamin B(12) analysis giving good recovery results. The surface morphologies of the SWCNT-chitosan PGE, chitosan modified PGE and unmodified PGE were characterized by using scanning electron microscopy (SEM). PMID- 21616650 TI - Investigation of the redox property of a metalloprotein layer self-assembled on various chemical linkers. AB - Myogloblin, a well-known metalloprotein, was immobilized on a gold surface using various chemical linkers to investigate the length effect of chemical linker on the electron transfer in protein layers, because chemical linkers play roles in the pathway that transfers the electron from the protein to the gold substrate and act as protein immobilization reagents. Chemical linkers with 2, 6, 11, and 16 carbons were utilized to confirm length-effects. The immobilization of protein and chemical linker was validated with surface plasmon resonance (SPR) and atomic force microscopy (AFM). The electrochemical property was evaluated by cyclic voltammetry (CV) and chronocoulometry (CC). In those results, redox peaks of immobilized protein were controlled via the length of chemical linkers, and it could be directly applied to the realization of bioelectronic device. PMID- 21616651 TI - [Varicella-associated purpura fulminans and deep vein thrombosis: a pediatric case report]. AB - Purpura fulminans (PF) and deep vein thrombosis are rare complications secondary to chicken pox disease. The presence of antibodies reflects an ongoing immunological process and requires specialized management. The present study reports a 4-year-old boy with no medical history who presented with purpura on the legs 10 days after chicken pox eruption. Laboratory tests showed a disseminated intravascular coagulation associated with low plasma protein C and S activities, and the presence of anti-protein S antibodies. A replacement therapy with protein C infusions and fresh frozen plasma was prescribed. The patient also underwent regular sessions of hyperbaric oxygen followed by the surgery. Fourteen days after the beginning of the purpuric lesions, he presented deep vein thrombosis (DVT) of the lower limbs and was treated with unfractionated heparin. This case report illustrates the pathophysiology of DVT occurring in a patient with chicken pox disease (i.e., acquired protein C and S deficiencies and anti protein S autoantibodies) and emphasizes the utility of thrombophilia testing in order to better adapt treatment. PMID- 21616652 TI - Glycosylation, galectins and cellular signaling. AB - Glycosylation is a common posttranslational modification of proteins and lipids of the secretory pathway that generates binding sites for galactose-specific lectins or galectins. Branching of Asn-linked (N-)glycans by the N acetylglucosaminyltransferases (Mgat genes) increases affinity for galectins. Both tissue-specific expression of the enzymes and the metabolic supply of sugar nucleotides to the ER and Golgi regulate glycan distribution while protein sequences specify NXS/T site multiplicity, providing metabolic and genetic contributions to galectin-glycoprotein interactions. Galectins cross-link glycoproteins forming dynamic microdomains or lattices that regulate various mediators of cell adhesion, migration, proliferation, survival and differentiation. There are a similar number of galactose-specific galectins in C. elegans and humans, but expression of higher-affinity branched N-glycans are a more recent feature of vertebrate evolution. Galectins might be considered a reading code for repetition of the minimal units of binding [Gal(NAc)beta1 3/4GlcNAc] and NXS/T site multiplicity in proteins. The rapidly evolving and structurally complex Golgi modifications to surface receptors are interpreted through affinity for the lattice, which regulates receptor levels as a function of the cellular environment, and thereby the probability of various cell fates. Many important questions remain concerning the regulation of the galectins, the glycan ligands and lattice interaction with other membrane domains and endocytic pathways. PMID- 21616653 TI - Utilization of municipal solid waste incineration fly ash for sulfoaluminate cement clinker production. AB - The feasibility of partially substituting raw materials with municipal solid waste incineration (MSWI) fly ash in sulfoaluminate cement (SAC) clinker production was investigated by X-ray diffraction (XRD), compressive strength and free expansion ratio testing. Three different leaching tests were used to assess the environmental impact of the produced material. Experimental results show that the replacement of MSWI fly ash could be taken up to 30% in the raw mixes. The good quality SAC clinkers are obtained by controlling the compositional parameters at alkalinity modulus (C(m)) around 1.05, alumina-sulfur ratio (P) around 2.5, alumina-silica ratio (N) around 2.0~3.0 and firing the raw mixes at 1250 degrees C for 2h. The compressive strengths of SAC are high in early age while that develop slowly in later age. Results also show that the expansive properties of SAC are strongly depended on the gypsum content. Leaching studies of toxic elements in the hydrated SAC-based system reveal that all the investigated elements are well bounded in the clinker minerals or immobilized by the hydration products. Although some limited positive results indicate that the SAC prepared from MSWI fly ash would present no immediate thread to the environment, the long-term toxicity leaching behavior needs to be further studied. PMID- 21616654 TI - Nanoparticle based DNA biosensor for tuberculosis detection using thermophilic helicase-dependent isothermal amplification. AB - The present study describes the development of a DNA based biosensor to detect Mycobacterium tuberculosis using thermophilic helicase-dependent isothermal amplification (tHDA) and dextrin coated gold nanoparticles (AuNPs) as electrochemical reporter. The biosensor is composed of gold nanoparticles (AuNPs) and amine-terminated magnetic particles (MPs) each functionalized with a different DNA probe that specifically hybridize with opposite ends of a fragment within the IS6110 gene, which is M. tuberculosis complex (MTC) specific. After hybridization, the formed complex (MP-target-AuNP) is magnetically separated from the solution and the AuNPs are electrochemically detected on a screen printed carbon electrode (SPCE) chip. The obtained detection limit is 0.01 ng/MUl of isothermally amplified target (105 bp). This biosensor system can be potentially implemented in peripheral laboratories with the use of a portable, handheld potentiostat. PMID- 21616655 TI - Methyl parathion hydrolase based nanocomposite biosensors for highly sensitive and selective determination of methyl parathion. AB - This article reports the fabrication of a nanocomposite biosensor for the sensitive and specific detection of methyl parathion. The nanocomposite sensing film was prepared via the formation of gold nanoparticles on silica particles, mixing with multiwall carbon nanotubes and subsequent covalent immobilization of methyl parathion hydrolase. The composite of the individual materials was finely tuned to offer the sensing film with high specific surface area and high conductivity. A significant synergistic effect of nanocomposites on the biosensor performance was observed in biosensing methyl parathion. The square wave voltammetric responses displayed well defined peaks, linearly proportional to the concentrations of methyl parathion in the range from 0.001 MUg mL-1 to 5.0 MUg mL 1 with a detection limit of 0.3 ng mL-1. The application of this biosensor in the analysis of spiked garlic samples was also evaluated. The proposed protocol can be used as a platform for the simple and fast construction of biosensors with good performance for the determination of enzyme-specific electroactive species. PMID- 21616656 TI - Ag(I)-cysteamine complex based electrochemical stripping immunoassay: ultrasensitive human IgG detection. AB - An ultrasensitive electrochemical immunosensor for a protein using a Ag (I) cysteamine complex (Ag-Cys) as a label was fabricated. The low detection of a protein was based on the electrochemical stripping of Ag from the adsorbed Ag-Cys complex on the gold nanoparticles (AuNPs) conjugated human immunoglobulin G (anti IgG) antibody (AuNPs-anti-IgG). The electrochemical immunosensor was fabricated by immobilizing anti-IgG antibody on a poly-5,2':5',2''-terthiophene-3' carboxylic acid (polyTTCA) film grown on the glassy carbon electrode through the covalent bond formation between amine groups of anti-IgG and carboxylic acid groups of polyTTCA. The target protein, IgG was sandwiched between the anti-IgG antibody that covalently attached onto the polyTTCA layer and AuNPs-anti-IgG. Using square wave voltammetry, well defined Ag stripping voltammograms were obtained for the each target concentration. Various experimental parameters were optimized and interference effects from other proteins were checked out. The immunosensor exhibited a wide dynamic range with the detection limit of 0.4 +/- 0.05 fg/mL. To evaluate the analytical reliability, the proposed immunosensor was applied to human IgG spiked serum samples and acceptable results were obtained indicating that the method can be readily extended to other bioaffinity assays of clinical or environmental significance. PMID- 21616657 TI - Comparison of relative potency of intrathecal bupivacaine for motor block in pregnant versus non-pregnant women. AB - BACKGROUND: Pregnancy is associated with facilitated spread of spinal and epidural anesthesia. There are limited data available for relative potency of motor block of neuraxial local anesthetics in non-pregnant versus pregnant women. The purpose of this study was to investigate the median effective dose (ED(50)) of intrathecal isobaric bupivacaine for motor block in non-pregnant and pregnant women and to estimate the respective potency ratio. METHODS: American Society of Anesthesiologists physical status I and II pregnant (n=35) and non-pregnant (n=35) patients undergoing elective cesarean delivery or elective gynecological surgery under combined spinal-epidural anesthesia were enrolled. According to the up-down sequential allocation technique, the dose of intrathecal isobaric bupivacaine for each patient was determined by the response of the previous patient in both groups. The initial dose of bupivacaine was 4 mg and the testing interval was set at 0.5 mg. Efficacy was determined by the occurrence of motor block in either lower limb as assessed by the modified Bromage scale within 5 min of spinal injection. RESULTS: The ED(50) of intrathecal bupivacaine for motor block was 4.51 (95% confidence interval (CI) 4.27-4.76) mg for non-pregnant women and 3.96 (95% CI 3.83-4.08) mg for pregnant women. The relative potency ratio for motor block for pregnant versus non-pregnant women was 1.14 (95% CI 1.05-1.24), (P=0.0037). CONCLUSIONS: Intrathecal bupivacaine was 1.14 times more potent for motor block in pregnant versus non-pregnant women. Our current data confirm the difference in local anesthetic requirement between non-pregnant and pregnant patients. PMID- 21616658 TI - CYP1A1*2A polymorphism as a predictor of clinical outcome in advanced lung cancer patients treated with EGFR-TKI and its combined effects with EGFR intron 1 (CA)n polymorphism. AB - BACKGROUND: Mutations in the epidermal growth factor receptor (EGFR) have been confirmed as predictors of efficacy for EGFR-tyrosine kinase inhibitors (TKIs). We investigated whether polymorphisms of the EGFR and cytochrome P450, family 1, member A1 (CYP1A1) genes were associated with clinical outcome in NSCLC patients treated with EGFR-TKI. METHODS: Genotypes for the intron 1 (CA)n repeat and R497K polymorphisms in the EGFR gene and the *2A (3801 T->C) and *2C (2455 A->G) polymorphisms in CYP1A1 gene were evaluated in 115 NSCLC patients by PCR-RFLP and DNA sequencing. Genetic polymorphisms were correlated with clinical outcomes of EGFR-TKIs. From a subgroup of patients whose tumour tissues were available, associations between somatic EGFR mutations, EGFR expression, and genomic polymorphisms were also analysed. RESULTS: EGFR intron 1 (CA)n and CYP1A1*2A polymorphisms were independent predictive factors (p=0.046, p=0.011, respectively) and the latter was also a prognostic factor (p=0.001) for patients treated with EGFR-TKIs. We also observed a strong synergistic effect from two genotypes. Specifically, patients with both the T/T allele of the CYP1A1 gene and shorter intron 1 CA repeats (<= 16 CA) of the EGFR gene showed an improved response (p=0.002) compared with patients with the T/C or C/C allele and longer intron 1 CA repeats (both alleles >16 CA). In contrast, for R497K and CYP1A1*2C, no relationship was observed with clinical outcome for patients treated with EGFR TKIs (p=0.573; p=0.629, respectively). Both SNPs in the CYP1A1 gene showed a correlation with EGFR somatic mutations. CONCLUSIONS: The findings of this study suggest that the CYP1A1*2A polymorphism is a predictor for clinical outcome in NSCLC patients treated with EGFR-TKI therapy, and combining analysis of both CYP1A1*2A and EGFR intron 1 (CA)n polymorphisms may be useful for predicting treatment outcome in NSCLC patients treated with EGFR-TKIs. PMID- 21616659 TI - Curcumin improves MMC-based chemotherapy by simultaneously sensitising cancer cells to MMC and reducing MMC-associated side-effects. AB - Mitomycin C (MMC) potently suppresses tumour growth. However, its use is limited by its severe toxicity to the kidney and bone marrow. The purpose of this study is to investigate whether the chemoprevention agent curcumin can reduce MMC associated side-effects and improve MMC efficacy in a breast cancer xenograft model. We first determined the effectiveness of combined MMC and curcumin to inhibit in vitro cell growth and to regress in vivo tumour outgrowth. We then investigated the mechanisms associated with MMC/curcumin-induced cell death by examining the effect of MMC/curcumin treatment on apoptosis, the activation of caspase-3, 8 and 9 and the expression of bcl-2 and bax. We also evaluated the ability of curcumin to alleviate MMC-associated side-effects by comparing the levels of creatinine/blood urea nitrogen (Cr/BUN) and glutamic oxalacetic transaminase/glutamic pyruvic transaminase (GPT/GOT) in serum between animals receiving MMC alone and MMC/curcumin. Curcumin significantly sensitised MCF-7 and MDA-MB-231 cells to MMC-induced cell death and improved MMC's ability to regress MCF-7 xenograft. MMC and curcumin together synergistically enhanced apoptosis in MCF-7 cells and the apoptosis most likely resulted from both the activation of caspases and modulation of bcl-2/bax expression. Most importantly, the inclusion of curcumin in MMC treatment decreased MMC-caused severe side-effects evidenced by significant improvement in the kidney function. Enhancing the tumoricidal effect of MMC, curcumin greatly reduces MMC-associated severe side-effects. Therefore, the combination treatment of MMC and curcumin may be of significant therapeutic benefit in treating breast cancer. PMID- 21616660 TI - A case of multiple brain metastases of uterine leiomyosarcoma with a literature review. AB - Brain metastasis from uterine leiomyosarcoma is extremely rare, and prognostically alarming despite various treatments. The authors report a case of multiple brain metastases from uterine leiomyosarcoma who took a favorable course after tumor resection and gamma-knife treatment. A 50-year-old woman with a history of hysterectomy for uterine leiomyosarcoma two years earlier, presented with a recent onset of headaches and vomiting. Multiple cerebral lesions were found by magnetic resonance imaging (MRI). The Karnofsky performance scale (KPS) was 40 with left hemiparesis and cerebellar ataxia. She was treated by resection of the left occipital and cerebellar tumors, followed by gamma-knife irradiation of the residual tumors. KPS was 70 at her discharge from the hospital. MRI failed to show recurrence of the intracranial lesions 6 months after irradiation. She remained at home until she died from massive intra-abdominal bleeding. This is the first case with multiple brain metastases from uterine leiomyosarcoma, who survived with remarkable neurological improvement for 12 months. No comparable survival has been reported in the literature. It is evident that surgical resection and additional gamma-knife irradiation contributed to early neurological recovery. PMID- 21616661 TI - Ethanol production from oil palm trunks treated with aqueous ammonia and cellulase. AB - Oil palm trunks are a possible lignocellulosic source for ethanol production. Low enzymatic digestibility of this type of material (11.9% of the theoretical glucose yield) makes pretreatment necessary. An enzymatic digestibility of 95.4% with insoluble solids recovery of 49.8% was achieved after soaking shredded oil palm trunks in ammonia under optimum conditions (80 degrees C, 1:12 solid-to liquid ratio, 8h and 7% (w/w) ammonia solution). Treatment with 60 FPU of commercial cellulase (Accellerase 1000) per gram of glucan and fermentation with Saccharomyces cerevisiae D(5)A resulted in an ethanol concentration of 13.3g/L and an ethanol yield of 78.3% (based on the theoretical maximum) after 96 h. These results indicate that oil palm trunks are a biomass feedstock that can be used for bioethanol production. PMID- 21616662 TI - Life cycle assessment of biogas infrastructure options on a regional scale. AB - A life cycle assessment has been completed of potential biogas infrastructures on a regional scale. Centralised and distributed infrastructures were considered along with biogas end uses of Combined Heat and Power (CHP) and injection to the gas grid for either transport fuel or domestic heating end uses. Damage orientated (endpoint) life cycle impact assessment method identified that CHP with 80% heat utilisation had the least environmental impact, followed by transport fuel use. Utilisation for domestic heating purposes via the gas grid was found to perform less well. A 32% difference in transportation requirement between the centralised and distributed infrastructures was found to have a relatively small effect on the overall environmental impact. Global warming impacts were significantly affected by changes in methane emissions at upgrading stage, highlighting the importance of minimising operational losses. PMID- 21616663 TI - Electricity generation in a membrane-less microbial fuel cell with down-flow feeding onto the cathode. AB - A novel membrane-less microbial fuel cell (MFC) with down-flow feeding was constructed to generate electricity. Wastewater was fed directly onto the cathode which was horizontally installed in the upper part of the MFC. Oxygen could be utilized readily from the air. The concentration of dissolved oxygen in the influent wastewater had little effect on the power generation. A saturation-type relationship was observed between the initial COD and the power generation. The influent flow rate could affect greatly the power density. Fed by the synthetic glucose wastewater with a COD value of 3500 mg/L at a flow rate of 4.0 mL/min, the developed MFC could produce a maximum power density of 37.4 mW/m(2). Its applicability was further evaluated by the treatment of brewery wastewater. The system could be scaled up readily due to its simple configuration, easy operation and relatively high power density. PMID- 21616664 TI - N,N'-Dialkylaminoalkylcarbonyl (DAAC) prodrugs and aminoalkylcarbonyl (AAC) prodrugs of 4-hydroxyacetanilide and naltrexone with improved skin permeation properties. AB - N,N'-Dialkylaminoalkylcarbonyl (DAAC) and aminoalkylcarbonyl (AAC) prodrugs of phenolic drugs acetaminophen (APAP) and naltrexone (NTX) are reported. The effects of incorporation of a basic amine group into the promoiety of an acyl prodrug of a phenolic drug on its skin permeation properties are also presented. DAAC-APAP prodrugs were synthesized via a three-step procedure starting with haloalkylcarbonyl esters which were reacted with five different amines: dimethylamine, diethylamine, dipropylamine, morpholine, and piperidine. The spacing between the amino group and the carbonyl group of the acyl group was 1-3 CH(2). After the hydrolysis of the ester, the carboxylic acid product was subsequently coupled with the parent drug via a dicyclohexyl carbodiimide (DCC) mediated coupling to yield the DAAC-APAP-HCl prodrugs in excellent yields. The AAC prodrugs were synthesized using commercially available Boc-protected amino acids using DCC or EDCI as coupling agents. The yields of the prodrugs synthesized using these two different methods have been compared. Half-lives (t(1/2)) of a few members of the DAAC and AAC series were measured in buffer (pH 6.0, 20mM). The members evaluated in hydrolysis experiments exhibit a t(1/2) range of 15-113min. Among AAC-APAP prodrugs, the isopropyl group in valinate-APAP HCl exerted a steric effect that increased the t(1/2) value for this prodrug compared to alaninate-APAP-HCl or prolinate-APAP-HCl. The 2-morpholinylacetate APAP prodrug was able to achieve twice the flux of APAP in in vitro diffusion cell experiments through hairless mouse skin. PMID- 21616665 TI - Triazoles as gamma-secretase modulators. AB - Synthesis, SAR, and evaluation of aryl triazoles as novel gamma secretase modulators (GSMs) are presented in this communication. Starting from the literature and in-house leads, we evaluated a range of five-membered heterocycles as replacements for olefins commonly found in non-acid GSMs. 1,2,3-C-aryl triazoles were identified as suitable replacements which exhibited good modulation of gamma-secretase activity, excellent pharmacokinetics and good central lowering of Abeta42 in Sprague-Dawley rats. PMID- 21616666 TI - Can biomechanical variables predict improvement in crouch gait? AB - Many patients respond positively to treatments for crouch gait, yet surgical outcomes are inconsistent and unpredictable. In this study, we developed a multivariable regression model to determine if biomechanical variables and other subject characteristics measured during a physical exam and gait analysis can predict which subjects with crouch gait will demonstrate improved knee kinematics on a follow-up gait analysis. We formulated the model and tested its performance by retrospectively analyzing 353 limbs of subjects who walked with crouch gait. The regression model was able to predict which subjects would demonstrate 'Improved' and 'Unimproved' knee kinematics with over 70% accuracy, and was able to explain approximately 49% of the variance in subjects' change in knee flexion between gait analyses. We found that improvement in stance phase knee flexion was positively associated with three variables that were drawn from knowledge about the biomechanical contributors to crouch gait: (i) adequate hamstrings lengths and velocities, possibly achieved via hamstrings lengthening surgery, (ii) normal tibial torsion, possibly achieved via tibial derotation osteotomy, and (iii) sufficient muscle strength. PMID- 21616667 TI - Are gait initiation parameters early markers of Huntington's disease in pre manifest mutation carriers? AB - Huntington's disease (HD) pre-manifest mutation carriers (PMCs) present early onset gait disturbances. Gait initiation encompasses the preparation and execution of the first step. By using paradigms with and without external cues, a gait initiation analysis can highlight the interaction between motor and cognitive aspects of movement preparation and execution. Hence, gait initiation disorders may constitute particularly interesting early markers of HD. The objective of the present study was to quantify gait initiation in PMCs. In a case control study, 17 PMCs (median age: 36.5) were compared with a group of 25 healthy controls (HCs, median age: 36) for gait initiation and a group of 57 HCs (median age: 38) for gait. Presymptomatic mutation carriers displayed a shorter first step duration and lower-amplitude postural adjustments. For the first step duration and speed, these impairments were more pronounced under self-triggered (ST) conditions. The PMCs displayed a lower gait speed, cadence and stride length and higher stride-to-stride variability. The latter parameter seemed capable of differentiating between PMCs and HCs with adequate sensitivity (0.81) and specificity (0.87). We confirmed the early-onset impairment of gait in general and first step execution in particular in PMCs (particularly under ST conditions). The temporal parameters of step execution (e.g. duration) and spatial parameters of postural adjustment (e.g. a backward shift in the centre of pressure) may be worth investigating as early markers of HD. However, two such parameters (stride-to-stride variability and first step duration under ST conditions) already appear to be sufficiently reliable diagnostic tools for differentiating between PMCs and HCs. PMID- 21616668 TI - Transverse plane rotation of the foot and transverse hip and pelvic kinematics in diplegic cerebral palsy. AB - External rotation of the foot associated with mid-foot break is a commonly observed gait abnormality in diplegic CP patients. Previous studies have shown a correlation between equinus and internal hip rotation in hemiplegic patients. This study aimed to determine if there was a correlation between the amount of transverse plane rotation in diplegic CP patients using kinematic data from standardised gait analysis. Lower limb data of 134 ambulant children with diplegic CP was analysed retrospectively determining the maximum change in foot, hip and pelvis rotation during loading response. Highly significant negative correlations (P=<0.001) were found between foot and hip movements and foot and pelvic movements. Equinus at initial contact diminished the foot:hip correlation while it enhanced the foot:pelvic correlation. There was less external rotation of the foot in equinus patients (P=0.012) and more external rotation of the pelvis in the equinus group (P=<0.001). This data reveal a correlation between transverse plane rotation at foot level to that at the hip and pelvis. The likely biomechanical explanation is relatively excessive transverse external rotation of the foot due to abnormalities such as mid-foot break. When under load, where the foot is fixed to the floor, internal rotation of the entire leg occurs. This is due to lever arm disease as a result of the relatively shortened foot and inefficiency of the plantar-flexion knee-extension couple. Equinus modulates the effect. When treating such patients, lever arm deformities at all levels must be considered to result in the best outcome and prevent recurrences. PMID- 21616669 TI - Postural balance in low back pain patients: Intra-session reliability of center of pressure on a portable force platform and of the one leg stand test. AB - Low back pain (LBP) patients have poorer postural control compared to healthy controls, and the importance of assessing and addressing balance is a matter of debate. In the clinic, balance is often tested by means of the one leg stand test (OLST) while research often employs center of pressure (CoP) on a force platform. Portable force platforms might be of clinical relevance, but their reliability for LBP patients in a clinical setting has not been demonstrated. As LBP patients are more dependent on vision compared to healthy controls, the ratio of tests performed with eyes open and eyes closed (Romberg Ratio) might be of clinical interest. This study aimed to assess postural balance in LBP patients by analyzing intra-session reliability of CoP parameters on a portable force platform, the Romberg Ratio, and the OLST. Furthermore, we aimed to determine whether CoP parameters and OLST measure identical aspects of postural stability. We examined 49 LBP patients and found acceptable reliability of the CoP parameters' trace length and velocity, whereas reliability regarding C90 area, the Romberg Ratio, and the OLST was poor. Correlations between the CoP parameters and OLST were insignificant. Reliability of trace length and velocity is acceptable and can be used as parameters when assessing CoP in LBP patients. PMID- 21616671 TI - Synthesis, structure-affinity relationships, and molecular modeling studies of novel pyrazolo[3,4-c]quinoline derivatives as adenosine receptor antagonists. AB - This paper reports the study of new 2-phenyl- and 2-methylpyrazolo[3,4-c]quinolin 4-ones (series A) and 4-amines (series B), designed as adenosine receptor (AR) antagonists. The synthesized compounds bear at the 6-position various groups, with different lipophilicity and steric hindrance, that were thought to increase human A(1) and A(2A) AR affinities and selectivities, with respect to those of the parent 6-unsubstituted compounds. In series A, this modification was not tolerated since it reduced AR affinity, while in series B it shifted the binding towards the hA(1) subtype. To rationalize the observed structure-affinity relationships, molecular docking studies at A(2A)AR-based homology models of the A(1) and A(3) ARs and at the A(2A)AR crystal structure were carried out. PMID- 21616672 TI - Extraction of tacit knowledge from large ADME data sets via pairwise analysis. AB - Pharmaceutical companies routinely collect data across multiple projects for common ADME endpoints. Although at the time of collection the data is intended for use in decision making within a specific project, knowledge can be gained by data mining the entire cross-project data set for patterns of structure-activity relationships (SAR) that may be applied to any project. One such data mining method is pairwise analysis. This method has the advantage of being able to identify small structural changes that lead to significant changes in activity. In this paper, we describe the process for full pairwise analysis of our high throughput ADME assays routinely used for compound discovery efforts at Pfizer (microsomal clearance, passive membrane permeability, P-gp efflux, and lipophilicity). We also describe multiple strategies for the application of these transforms in a prospective manner during compound design. Finally, a detailed analysis of the activity patterns in pairs of compounds that share the same molecular transformation reveals multiple types of transforms from an SAR perspective. These include bioisosteres, additives, multiplicatives, and a type we call switches as they act to either turn on or turn off an activity. PMID- 21616670 TI - Mass spectrometry accelerates membrane protein analysis. AB - Cellular membranes are composed of proteins and glyco- and phospholipids and play an indispensible role in maintaining cellular integrity and homeostasis, by physically restricting biochemical processes within cells and providing protection. Membrane proteins perform many essential functions, which include operating as transporters, adhesion-anchors, receptors, and enzymes. Recent advancements in proteomic mass spectrometry have resulted in substantial progress towards the determination of the plasma membrane (PM) proteome, resolution of membrane protein topology, establishment of numerous receptor protein complexes, identification of ligand-receptor pairs, and the elucidation of signaling networks originating at the PM. Here, we discuss the recent accelerated success of discovery-based proteomic pipelines for the establishment of a complete membrane proteome. PMID- 21616673 TI - Co-localization of PCNA, VCAM-1 and caspase-3 with nestin in xenografts derived from human anaplastic astrocytoma and glioblastoma multiforme tumor spheres. AB - The cancer stem cell hypothesis proposes that tumors contain a small subset of cancer cells, the cancer stem cells, which constitute a reservoir of self sustaining cells with the exclusive ability to self-renew and maintain the tumor. Markers that define cancer stem cells that are capable of recapitulating brain tumors as xenografts in mice has not been described. We investigated the relationship between expression of nestin and that of PCNA, VCAM-1 and caspase-3 in the xenografts developed from human anaplastic astrocytoma and glioblastoma tumor-derived spheres in the brain of nude mouse. Xenografts obtained from astrocytoma tumor stem cells (ATSC) and glioblastoma tumor stem cells (GTSC) have showed a large number of cells positive for both PCNA and the nestin, demonstrating that nestin expressing cells have a high rate of proliferation. Xenografts from GTSC showed heterogeneous staining pattern with cells that express both nestin and VCAM-1, whereas others cells remained complete negative. In this case it was noticed that most tumor cells with large or multinucleated nuclei coexpress nestin and VCAM-1. In xenografts from ATSC most cells positive for nestin express VCAM-1 and in this case the two proteins appear to occupy the same cytoplasmic region. Both GTSC and ATSC derived xenografts showed cells positive for both caspase-3 and for nestin detected mainly as single cells and as cell clusters located near or around a blood vessel. PMID- 21616674 TI - Pulmonary resection for lung cancer with malignant pleural disease first detected at thoracotomy. AB - OBJECTIVES: The prognosis of non-small-cell lung cancer (NSCLC) patients with malignant pleural disease (MPD), characterized by malignant pleural effusion and/or malignant pleural nodules, is reported to be poor, and patients with MPD are generally not subjected to surgery. However, whether or not the primary tumor should be resected, when MPD is first detected at thoracotomy, is controversial. METHODS: The clinical records of 1623 consecutive NSCLC patients, who underwent surgery between 1990 and 2007, were retrospectively reviewed. A hundred patients (6.2%) were classified with pathological stage IV disease according to the seventh edition of the Union for International Cancer Control (UICC) staging system. There were 73 patients with MPD, which included 32 with effusion without nodules (MPE) and 41 with nodules with or without effusion (MPN). Intra- or postoperative pleural chemotherapy was administered to 37 MPD patients. RESULTS: The median survival time, the 3-year survival rate and the 5-year survival rate for MPD patients were 25.9 months, 41.4%, and 23.7%, respectively, which are better outcomes than those for M1b patients (8.7 months, 18% and 18%, respectively) (log-lank test: p=0.014). Among MPD patients, N0-1 disease was determined to be a favorable prognostic factor (p=0.01). MPD status (MPE or MPN) was not prognostically significant (p=0.40). MPE patients with N0-1 disease had a significantly better prognosis with a 5-year survival rate of 63.6% compared to MPE patients with N2-3 disease (p=0.003). Twenty-seven percent of MPN patients with N0-1 disease achieved 5-year survival, whereas none of the MPD patients with N2-3 disease survived longer than 5 years after surgery. CONCLUSIONS: The prognosis of patients with surgically detected MPD, who underwent resection, was better than that of M1b patients. MPE patients with N0-1 disease may be candidates for resection. PMID- 21616675 TI - Mild-to-moderate hypothermia in aortic arch surgery using circulatory arrest: a change of paradigm? AB - OBJECTIVES: Antegrade cerebral perfusion makes deep hypothermia non-essential for neuroprotection; therefore, there is a growing tendency to increase the body temperature during circulatory arrest with selective brain perfusion. However, very little is known about the clinical efficacy of mild-to-moderate hypothermia for ischemic organ protection during circulatory arrest. The aim of this study was to evaluate the safety and efficiency of mild-to-moderate hypothermia for lower-body protection during aortic arch surgery with circulatory arrest and antegrade cerebral perfusion. METHODS: Between January 2005 and December 2009, a total of 347 patients underwent non-emergent arch surgery. In all patients, the systematic cooling was adapted to the expected time of circulatory arrest, and cerebral perfusion was performed at a constant blood temperature of 28 degrees C. There were 40 cardiac or aortic re-operations, 312 patients had concomitant aortic valve or root surgery, and 10 patients had replacement of the descending aorta. All examined data were collected prospectively. RESULTS: The duration of circulatory arrest and the deepest rectal temperature were 18+/-11 min (range, 6 70 min) and 31.5+/-1.6 degrees C (range, 26.0-35.0 degrees C) for all 347 patients, and 34+/-12 min (range, 17-70 min) and 29.9+/-1.7 degrees C (range, 26.0-34.6 degrees C) for 77 patients having total/subtotal arch replacement. The maximum serum lactate level on the first postoperative day was, on average, 2.3+/ 1.2 mmol l(-1). In the statistical analysis, no association between the duration of temperature-adapted circulatory arrest and lactate, creatinine, or lactate dehydrogenase levels after surgery could be demonstrated. The 30-day mortality was 0.9%. Permanent neurological deficit or temporary dysfunction occurred in three (0.9%) and eight (2.3%) patients, respectively. No paraplegia and no hepatic failure were reported; however, mesenteric ischemia occurred in one patient with severe stenosis of the celiac and upper mesenteric arteries. Temporary dialysis was necessary primarily after surgery in five patients. All of them underwent hemiarch replacement only, and four patients had an increased creatinine level before surgery. CONCLUSION: Systemic mild-to-moderate hypothermia that is adapted to the duration of circulatory arrest is a simple, safe, and effective method of organ protection and can be recommended in routine aortic arch surgery with circulatory arrest and cerebral perfusion. PMID- 21616676 TI - Urgent lung transplantation in cystic fibrosis patients: experience of a French center. AB - OBJECTIVE: We report preliminary results obtained with urgent lung transplantation (ULTx) in cystic fibrosis (CF) patients, based on a French high emergency lung allocation (HELA) system, and the impact of this system on waiting list death. METHODS: The medical records of the first 15 CF patients receiving ULTx between June 2007 and May 2010 at Hopital Europeen Georges Pompidou, France, were retrospectively reviewed. ULTx patients (URG group, n=15) were compared with our entire cohort of CF patients receiving elective lung transplants (LTx) (ELT group, n=118). RESULTS: Both groups were similar in terms of use of cardiopulmonary bypass (CPB), length of stay in the intensive care unit (ICU), and intubation > 72 h. Incidence of primary graft dysfunction (PGD) and perioperative mortality was also similar in both groups, but graft ischemic time and severity of PGD were higher in the URG group. One-year and 2.5-year survival rates were, respectively, 73% and 54.5% for the URG group. Death on the waiting list and time to LTx (including all pulmonary diagnoses) decreased by 67% and 64%, respectively. CONCLUSIONS: Although still preliminary and with a short follow-up period, our results suggest that the allocation of LTx to CF patients based on the HELA criteria yielded acceptable outcomes and improved waiting-list death rate and time to LTx. PMID- 21616677 TI - Neoadjuvant therapy in early-stage breast cancer. AB - Neoadjuvant (preoperative) chemotherapy is becoming a commonly used option for women with early-stage breast cancer, allowing a greater proportion of patients to undergo breast conservation surgery. Neoadjuvant chemotherapy also allows the early assessment of response or resistance to chemotherapy and facilitates chemotherapy delivery prior to any surgical alterations to the vasculature. Ongoing research is examining the potential benefits of neoadjuvant therapy with cytotoxic agents as well as other treatments, including endocrine therapies and biologic agents. Additionally, biomarkers are being intensively investigated as methods for identifying patients who will most likely benefit from neoadjuvant chemotherapy. As of yet, neoadjuvant chemotherapy has not demonstrated a definitive benefit over adjuvant (postoperative) chemotherapy with regard to prolonging survival. It remains to be seen whether novel cytotoxic agents used in the neoadjuvant setting will improve pathologic clinical response rates and ultimately improve long-term outcome in women with early-stage breast cancer. PMID- 21616679 TI - Association of TNF-alpha promoter gene G-308A polymorphism with metabolic syndrome, insulin resistance, serum TNF-alpha and leptin levels in Indian adult women. AB - BACKGROUND: Tumour necrosis factor alpha is a multifunctional proinflammatory cytokine involved in the pathogenesis of metabolic syndrome, insulin resistance, and obesity. Aim of this study is to investigate in a North Indian female population the impact of the G-308A TNF-alpha variant on various components of the metabolic syndrome, Insulin Resistance, serum TNF-alpha and Leptin levels. METHODS: The G-308A TNF-alpha polymorphism has been studied in 269 females with metabolic syndrome (NCEP ATP III criteria) (age 31.91+/-6.05) and 272 healthy females without metabolic syndrome (age 30.96+/-7.01). The G-308A variant was detected by PCR amplification and Nco-1 digestion. RESULTS: Homozygous mutant genotype (AA) (p=<0.001: OR=3.24: 95% CI=2.15-4.89) and mutant allele (A) (p=<0.001: OR=3.04: 95% CI=2.08-4.43) of TNF-alpha was significantly less frequently observed in the control population as compared to study group. Furthermore, on dividing the subjects into two groups according to the absence (TNF-1 allele) or presence of the mutant A (TNF-2) allele, significant results were obtained in most of the metabolic risk factors. CONCLUSIONS: Our results suggest that the G-308A polymorphism of the TNF-alpha gene may be independently associated with hypertension, leptin level and hypercholesterolemia leading to metabolic syndrome independent of Insulin resistance and hyperglycemia. PMID- 21616678 TI - Increased very low density lipoprotein (VLDL) secretion, hepatic steatosis, and insulin resistance. AB - Insulin resistance (IR) affects not only the regulation of carbohydrate metabolism but all aspects of lipid and lipoprotein metabolism. IR is associated with increased secretion of VLDL and increased plasma triglycerides, as well as with hepatic steatosis, despite the increased VLDL secretion. Here we link IR with increased VLDL secretion and hepatic steatosis at both the physiologic and molecular levels. Increased VLDL secretion, together with the downstream effects on high density lipoprotein (HDL) cholesterol and low density lipoprotein (LDL) size, is proatherogenic. Hepatic steatosis is a risk factor for steatohepatitis and cirrhosis. Understanding the complex inter-relationships between IR and these abnormalities of liver lipid homeostasis will provide insights relevant to new therapies for these increasing clinical problems. PMID- 21616681 TI - Population pharmacokinetics of lidocaine administered during and after cardiac surgery. AB - OBJECTIVE: The objective of this study was to determine the pharmacokinetics of lidocaine in a 48-hour infusion in patients undergoing cardiac surgery with cardiopulmonary bypass (CPB). DESIGN: A retrospective substudy of a clinical trial assessing the efficacy of intravenous lidocaine for postoperative cognitive decline. SETTING: A university hospital. PARTICIPANTS: Ninety-nine patients undergoing cardiac surgery with CPB. INTERVENTIONS: After the induction of anesthesia, lidocaine was administered as a bolus of 1 mg/kg and followed by a continuous infusion at 4 mg/min for the 1st hour, 2 mg/min for the 2nd hour, and 1 mg/min for the next 46 hours. MEASUREMENTS AND MAIN RESULTS: Blood samples were taken at baseline, the end of CPB, and 24 and 48 hours after CPB for the measurement of the plasma concentration of lidocaine. Lidocaine levels increased significantly over time despite a constant rate of infusion (p < 0.05). The pharmacokinetics of lidocaine was best described by a 2-compartment model, and body weight was found to be a significant factor for the volume of the central compartment and clearance. The final pharmacokinetic parameters were V(1)(L) = 0.0619*weight, V(2)(L) = 187, CL(1) (L/min) = 0.00419*weight, and CL(2) (L/min) = 8.92. CONCLUSIONS: A 2-compartment pharmacokinetic model best describes the plasma concentrations of a 48-hour lidocaine infusion in patients undergoing cardiac surgery with CPB. The inclusion of body weight as a covariate on clearance and central compartment improves the model. Lidocaine infusions should be dosed by body weight and decreased after 24 hours to avoid potential toxicity in long-term infusions. PMID- 21616682 TI - The prognostic role of electrocorticography in tailored temporal lobe surgery. AB - Intraoperative electrocorticography (ECoG) has been in clinical use for many decades, yet the validity of this procedure in guiding resective surgery for temporal lobe epilepsy (TLE) is still uncertain, especially in tailored temporal lobectomies in cases of TLE with hippocampal sclerosis. METHODOLOGY: Using a case control design combined with descriptive and comparative analyses we retrospectively evaluated two groups of patients: patients (n=20) who had tailored temporal lobectomies guided by intraoperative ECoG and patients (n=19) who had standard temporal lobectomies without ECoG. Clinical and neuroimaging data, pre- and post-resection ECoG recordings, and seizure-free outcomes were reviewed. RESULTS: Of the 20 patients who underwent epilepsy surgery guided by ECoG, pre-resection ECoG studies found spikes both in mesial temporal lobe (MTL) and lateral temporal lobe (LTL) in 10 patients, and restricted to the LTL or MTL in 7 and 3 patients respectively. Postsurgical ECoG captured residual epileptiform activity in only 3 patients, all of whom had MTL and LTL spikes prior to surgery. Postsurgical follow-up at 16.3 (+/-6.7) months found 15 (75%) patients were seizure free (1A), while 5 (25%) had other outcomes. Analysis found no difference in the proportion of seizure-free outcomes between patients with residual epileptiform activity compared to those without residual epileptiform activity, or between patients who had a tailored resection guided by ECoG and patients who had standard resections without ECoG monitoring. CONCLUSION: The patients who underwent tailored temporal lobe epilepsy surgery guided by ECoG had similar outcome compared with the patients with epilepsy surgery not guided by ECoG. PMID- 21616683 TI - Mycetoma or synovial sarcoma? A case report with review of the literature. AB - Mycetoma, also commonly referred to as Madura foot, is statistically rare in the United States. However, it is endemic to other parts of the world. It is a pseudotumor characterized by a triad of tumefaction, draining sinuses, and grains. Two types exist, with each caused by different groups of organisms that require different treatment approaches. Therefore, the exact diagnosis and culture of the organism is vital to successful treatment outcomes. Synovial sarcoma, in contrast, is a malignancy much more commonly seen in the United States. It is characterized by a well-circumscribed, often palpable, mass that is usually well delineated on magnetic resonance imaging. It has characteristic histologic and genetic features that help distinguish it from other soft tissue masses. We present a case of a soft tissue mass diagnosed in the United States. The patient had several clinical and radiographic features of synovial sarcoma but the histologic outcome was mycetoma. The case is followed by a review of the published data. PMID- 21616684 TI - Reconstructive options for defects after melanoma excision in the foot and ankle region. AB - Cutaneous wounds created by excision of primary cutaneous melanoma localized to the foot and ankle can be challenging to reconstruct. A wide range of coverage techniques are available for wound coverage. The techniques we have found useful in this regard include skin grafts and flaps, as well as specialized wound dressings. In this techniques report, we describe the specialized anatomic surface subunits of the foot and ankle and the reconstructive coverage methods we have found useful for each site. PMID- 21616680 TI - Costimulatory pathways in transplantation. AB - Secondary, so-called costimulatory, signals are critically required for the process of T cell activation. Since landmark studies defined that T cells receiving a T cell receptor signal without a costimulatory signal, are tolerized in vitro, the investigation of T cell costimulation has attracted intense interest. Early studies demonstrated that interrupting T cell costimulation allows attenuation of the alloresponse, which is particularly difficult to modulate due to the clone size of alloreactive T cells. The understanding of costimulation has since evolved substantially and now encompasses not only positive signals involved in T cell activation but also negative signals inhibiting T cell activation and promoting T cell tolerance. Costimulation blockade has been used effectively for the induction of tolerance in rodent models of transplantation, but turned out to be less potent in large animals and humans. In this overview we will discuss the evolution of the concept of T cell costimulation, the potential of 'classical' and newly identified costimulation pathways as therapeutic targets for organ transplantation as well as progress towards clinical application of the first costimulation blocking compound. PMID- 21616685 TI - Giant cell tumor of the distal phalanx of the biphalangeal fifth toe: a case report and review of the literature. AB - Giant cell tumor of the distal phalanx of a toe is rather unusual. We report an unusual case of giant cell tumor arising at the distal phalanx of the left biphalangeal fifth toe in a 13-year-old boy. This was treated successfully with curettage and packing with bone substitution. PMID- 21616687 TI - Lateral leg compartment syndrome caused by ill-fitting compression stocking placed for deep vein thrombosis prophylaxis during surgery: a case report. AB - Isolated lateral leg compartment syndrome is a relatively rare event, with potentially devastating consequences. We present a case of a 44-year-old man with isolated lateral leg compartment syndrome caused by a compression stocking used for deep vein thrombosis prophylaxis during surgery. It was found to be excessively tight around the patient's proximal calf postoperatively. He underwent lateral leg compartment fasciotomy and delayed wound closure with a split-thickness skin graft. At the 7-month follow-up visit, he had returned to full activity, had no muscle deficits, and had numbness in the distribution of the intermediate dorsal cutaneous nerve. PMID- 21616686 TI - Outcome of subtalar fusion using bovine cancellous bone graft: a retrospective case series. AB - Solvent preserved bovine cancellous bone graft (Tutobone((r))) has been promoted as an alternative to autologous bone graft. The aim of our study was to compare the outcomes of subtalar fusion in patients in whom Tutobone((r)) was used with the outcomes in patients in whom it was not used. This was a retrospective comparative study. Tutobone((r)) was used in 9 patients in the test group. Of these repairs, 6 were isolated subtalar fusions, and 3 were performed as a part of triple arthrodesis. A total of 17 patients were included in the control group; 4 underwent autologous iliac crest grafting and 13 received a local bone graft from excised joint surfaces. At 12 months after surgery, 8 of the 9 in the Tutobone((r)) group had persistent pain and radiologic signs of nonunion confirmed on computed tomography scan. All 17 in the other group had successful clinical and radiologic fusion at 12 months. We believe this is sufficient evidence to advise against the use of bovine cancellous bone graft material for subtalar fusion surgery. PMID- 21616688 TI - Foot and shoe size mismatch in three different New York City populations. AB - Proper shoe size is an important element of foot health, especially in the elderly and diabetic populations. An improper fit can lead to pain, functional limitations, and falls. The aim of the present study was to determine the proportion of adults who are unaware of their own shoe size in 3 different New York City populations: a foot specialist private practice, an academic diabetic foot and ankle clinic, and a charity care center, the Bowery clinic, serving the homeless. A shoe size mismatch was defined as a difference of at least 0.5 in size between the measured foot and the shoe size. Demographic data were collected during the examination and retrospectively by chart review. A total of 235 volunteers participated in our study. A significant difference in the prevalence of the measured foot and shoe size mismatch was found between the cohort from the private practice compared with both the diabetic foot and ankle clinic and the Bowery clinic (P < .01 and P < .01, respectively). A significant difference was also detected (P < .05) between the private practice and the Bowery mission cohort when a difference of at least 1.5 sizes was present between the measured foot and the shoe size. Of those with a foot to shoe size mismatch, 60% had a difference of more than 0.5 in the shoe size between their right and left foot. In conclusion, our findings suggest that proper footwear sizing is lacking among a large proportion of our patients and that an adequate shoe size can be achieved with proper counseling. PMID- 21616689 TI - Subextensor digitorum brevis abscess after blunt trauma. AB - It is unusual for an abscess to form after direct blunt trauma without a penetrating injury. There are several reports of submuscular or intramuscular abscess formation after blunt trauma; however, none has been reported in the foot. We present an interesting case of a patient who presented after a fairly benign, low energy injury to the foot. The patient was found to have a submuscular abscess that was successfully managed with a surgical incision and drainage. PMID- 21616690 TI - Bilateral clubfoot in three homozygous preterm triplets. AB - The etiology of congenital idiopathic talipes equinovarus deformity is unclear. Studies on populations, families, and twins have suggested a genetic component. However, the mode of inheritance does not fit classic patterns. The intrauterine posture and environmental and developmental causative factors have also been associated with the deformity. Neurologic, muscular, bony, connective tissue, and vascular structures can be affected. We present the case of monochorionic triplets with bilateral congenital idiopathic talipes equinovarus deformities. To the best of our knowledge, such a presentation has not been previously described and supports a genetic etiology of congenital idiopathic talipes equinovarus deformity. PMID- 21616691 TI - Active surveillance vs. treatment for low-risk prostate cancer: a cost comparison. AB - OBJECTIVE: Radical prostatectomy (RP) and radiation therapy are standard curative approaches for low-risk prostate cancer (PC). Active surveillance (AS) is becoming an increasingly accepted management alternative for low-risk PC. Our aim is to compare the cumulative medical costs of treatment vs. AS. METHODS AND MATERIALS: We collected data on the cumulative medical costs of open radical retropubic prostatectomy (RRP), robotic-assisted radical prostatectomy (RARP), external beam radiotherapy (EBRT), brachytherapy (BT), and AS at our institution. For physicians' reimbursements, Medicare values of our region were used to maintain uniformity. For inpatient costs other than reimbursements, we used the mean cost at our institution. The costs of RRP and RARP involve preoperative investigations, medical clearance, physicians' fees, inpatient costs, and pathologic examination of prostatectomy specimen and follow-up. The inpatient costs include the operating room, disposable equipment, anesthesia, post anesthesia care, transfusion, and hospital stay. The cost of EBRT involves the cost of consultation, planning, simulation and treatment sessions, and follow-up. BT costs involved radiotherapy planning as well as inpatients costs. AS protocol involves regular visits, transrectal ultrasound guided biopsies, prostate specific antigen (PSA) testing. To evaluate the cost of treating complications, treatment after AS, and treatment for recurrence, we created a Markov model based on recent studies and our experience. RESULTS: The cumulative costs of RRP are $9,732 (1 year), $10,360 (2 years), $12,209 (5 years), and $15,084 (10 years). While for RARP, the costs are $17,824 (1 year), $18,308 (2 years), $20,117 (5 years), and $22,762 (10 years). The costs of EBRT are $20,730 (1 year), $20,969 (2 years), $22,043 (5 years), and $23,953 (10 years). BT costs are $14,061 (1 year), $14,300 (2 years), $15,374 (5 years), and $17,284 (10 years). The costs of AS are $1,154 (1 year), $2,308 (2 years), $8,761 (5 years), and $13,116 (10 years). CONCLUSIONS: The cumulative medical costs of RARP and EBRT are much higher than BT, RRP, and AS. AS is associated with a different cost distribution in which the initial cost is low and relatively higher cost of follow-up. Despite the higher follow-up cost, AS remains the most cost effective alternative for low risk PC. PMID- 21616692 TI - A potential pathway for managing diabetic patients with arterial emboli detected by retinal screening. AB - OBJECTIVE: The aim was to review a pathway of care for diabetic patients found to have retinal artery emboli detected by retinal screening. DESIGN: This was a retrospective review of a pathway agreed in 2001 by a multidisciplinary team. MATERIALS AND METHODS: The prospectively collected Gloucestershire Diabetic Retinal Screening Programme database was reviewed; patients sent for carotid duplex imaging underwent review of their scan results and their casenotes. RESULTS: The prevalence of retinal emboli was 214 out of 25,299 diabetic patients who had retinal screening (0.85%). Some 200 diabetic patients underwent carotid duplex imaging; 23 had ipsilateral and 2 had contralateral carotid stenosis > 70%. Of these, ten patients underwent carotid endarterectomy without any major morbidity. CONCLUSIONS: [corrected] A pathway was established for the small number of patients with retinal emboli, and could be tested in other populations. PMID- 21616693 TI - Cyclic alternating pattern (CAP): the marker of sleep instability. AB - Cyclic alternating pattern CAP is the EEG marker of unstable sleep, a concept which is poorly appreciated among the metrics of sleep physiology. Besides, duration, depth and continuity, sleep restorative properties depend on the capacity of the brain to create periods of sustained stable sleep. This issue is not confined only to the EEG activities but reverberates upon the ongoing autonomic activity and behavioral functions, which are mutually entrained in a synchronized oscillation. CAP can be identified both in adult and children sleep and therefore represents a sensitive tool for the investigation of sleep disorders across the lifespan. The present review illustrates the story of CAP in the last 25 years, the standardized scoring criteria, the basic physiological properties and how the dimension of sleep instability has provided new insight into pathophysiolology and management of sleep disorders. PMID- 21616694 TI - From analytic inversion to contemporary IMRT optimization: radiation therapy planning revisited from a mathematical perspective. AB - In this paper we look at the development of radiation therapy treatment planning from a mathematical point of view. Historically, planning for Intensity-Modulated Radiation Therapy (IMRT) has been considered as an inverse problem. We discuss first the two fundamental approaches that have been investigated to solve this inverse problem: Continuous analytic inversion techniques on one hand, and fully discretized algebraic methods on the other hand. In the second part of the paper, we review another fundamental question which has been subject to debate from the beginning of IMRT until the present day: The rotation therapy approach versus fixed angle IMRT. This builds a bridge from historic work on IMRT planning to contemporary research in the context of Intensity-Modulated Arc Therapy (IMAT). PMID- 21616695 TI - Healthcare-associated pneumonia: diagnostic criteria and distinction from community-acquired pneumonia. AB - BACKGROUND: Traditionally, pneumonia developing in patients who receive healthcare services in the outpatient environment has been classified as community-acquired pneumonia (CAP). However, recent investigations suggest that this type of infection, known as healthcare-associated pneumonia (HCAP), is distinct from CAP in terms of its epidemiology, etiology, and risk for infection with multidrug-resistant (MDR) pathogens. METHODS: A Medline literature review of available clinical studies using the term HCAP was conducted to determine outcomes compared to CAP and effective empiric treatment strategies. RESULTS: Analysis of multi-institutional clinical data showed that mortality in hospitalized patients with HCAP is greater than that in CAP, and patients with HCAP received inappropriate initial empiric antibiotic treatment more frequently than CAP patients. The bacterial pathogens associated with HCAP also differed from CAP with potentially MDR Gram-positive and Gram-negative bacteria being more common in HCAP. CONCLUSIONS: All patients hospitalized with suspected HCAP should be evaluated for their underlying risk of infection with MDR pathogens. Because HCAP is similar to hospital-acquired pneumonia (HAP), both clinically and etiologically, it should be treated as HAP until culture data become available. PMID- 21616697 TI - [Arthroscopic treatment for Preiser's disease by partial resection of the scaphoid and pyrocarbone's implant, preliminary results: a case report and literature review]. AB - Idiopathic avascular necrosis of the scaphoid, also called Preiser's disease, has been diagnosed in only 150 cases. Its clinical and radiological description is well established, but there is no consensus regarding its treatment. We report the case of a 53-year-old woman, with an advanced disease; treated arthroscopically by partial resection of the scaphoid, and insertion of an adapted pyrocarbon implant. Preliminary results at 6 months were satisfying in terms of pain. However, there was little improvement in range of motion, with a final range of 150 degrees . PMID- 21616698 TI - A computational modeling approach of the jet-like acoustic streaming and heat generation induced by low frequency high power ultrasonic horn reactors. AB - High power ultrasound reactors have gained a lot of interest in the food industry given the effects that can arise from ultrasonic-induced cavitation in liquid foods. However, most of the new food processing developments have been based on empirical approaches. Thus, there is a need for mathematical models which help to understand, optimize, and scale up ultrasonic reactors. In this work, a computational fluid dynamics (CFD) model was developed to predict the acoustic streaming and induced heat generated by an ultrasonic horn reactor. In the model it is assumed that the horn tip is a fluid inlet, where a turbulent jet flow is injected into the vessel. The hydrodynamic momentum rate of the incoming jet is assumed to be equal to the total acoustic momentum rate emitted by the acoustic power source. CFD velocity predictions show excellent agreement with the experimental data for power densities higher than W(0)/V >= 25kWm(-3). This model successfully describes hydrodynamic fields (streaming) generated by low-frequency high-power ultrasound. PMID- 21616696 TI - Outpatient follow-up of patients hospitalized for acute leptospirosis. AB - OBJECTIVE: The outcome of leptospirosis after the resolution of acute disease, either spontaneously or after treatment, is not well described. The aim of this study was to assess the possible sequelae of acute leptospirosis after hospital discharge. METHODS: We report here a prospective study carried out in Sao Paulo, Brazil in which patients hospitalized for leptospirosis were followed in the outpatient setting. RESULTS: Forty-seven patients were serially assessed: 32 severe and 15 mild cases. Early and late complications were not common in either group, but subjective complaints were common in the first few weeks after hospital discharge (53% of severe cases, 40% of mild cases). Two patients had continuing complaints: one had profound general malaise and the other developed new onset panic disorder. The sample analyzed represented 26% of the patients hospitalized with leptospirosis in the city of Sao Paulo during the study period. The duration of follow-up was an average of approximately 20 days at the first visit, and approximately 40 days at the second visit. Forty-seven patients came for one follow-up visit and 22 of the same patients had two follow-up visits. CONCLUSIONS: While two of 47 patients reported continuing symptoms after hospitalization for acute leptospirosis, no definitive, objective evidence of chronic sequelae due to this infection was proven. While preliminary, these observations point to the need for a prospective, rigorous and systematic study to definitively determine and characterize late complications and chronic disease after acute leptospirosis. PMID- 21616699 TI - A new parameter for characterizing balancing ability on an unstable oscillatory platform. AB - An oscillatory platform can be used as a simple measuring instrument for modelling balancing capacity. We sought (1) to determine the test-retest reliability of the Lehr's damping ratio for characterizing the balancing ability on an unstable oscillatory platform of 20 healthy young and 20 healthy elderly participants and (2) to compare balancing ability on an unstable oscillatory platform between groups. Tests were performed during standing on both limbs and on one limb (dominant or non-dominant). The average of differences of the Lehr's damping ratio was 0.004+/-0.003 (3.77%+/-3.89%), and tests conducted at different times showed strong correlations (range 0.978-0.998). The average value of the Lehr's damping ratio was significantly smaller for elderly compared to young participants and it was significantly smaller for participants standing on their non-dominant limb compared to when they stood on both limbs or on the dominant limb. Based on this reproducible measuring method, Lehr's damping ratio depends on lateral dominance, and age. PMID- 21616700 TI - A practical technique for measuring human biofluid conductivity using high gain frequency characteristics. AB - Currently, the study of ion composition and performance in human biofluids plays an important role in biomedical engineering research and technology. This field may become universal for human diagnostics; it allows early detection of different diseases in humans by measuring changes in ion behaviour in human biofluids. Practical experiments were conducted to analyse the liquid composite electrolyte conductivity in an alternating electric current field. These experiments allow the contribution of separate types of ions to the overall conductivity to be estimated. The method of estimating the concentration of active ions contained in biofluids is also introduced; it illustrates the possibility of performing qualitative and quantitative analysis over a wide range of concentrations and compositions. The authors present a procedure to determine the concentration of active liquid ions based on conductivity gain-frequency characteristic curve tracing. The experimental results validate the practical use of the proposed method. The results of this research are promising, and further investigation is required to further improve the method. PMID- 21616701 TI - Migrants and healthcare: investigating patient mobility among migrants in Ireland. AB - Drawing on detailed interviews with 60 recent migrants to Ireland, we discuss the extent and nature of patient mobility. The paper is framed by the typology of patient mobility outlined by Glinos et al. (2010), which highlights patient motivation and funding. We pay particular attention to four key areas: availability of health care for migrants living in Ireland; affordability of care as a push factor for patient mobility; how migrants' perceptions of care affect their decision about where to avail of care; and the impact of familiarity on patient mobility. We provide empirical support for this typology. However, our research also highlights the fact that two factors - availability and familiarity - require further elaboration. Our research demonstrates the need for greater levels of awareness of culture specificity on the part of both migrants and healthcare providers. It also highlights the need to investigate the social and spatial activities of migrants seeking health care, both within and beyond national boundaries. PMID- 21616702 TI - The role of testosterone in social interaction. AB - Although animal researchers established the role of testosterone as a 'social hormone' decades ago, the investigation of its causal influence on human social behaviors has only recently begun. Here, we review and discuss recent studies showing the causal effects of testosterone on social interactions in animals and humans, and outline the basic neurobiological mechanisms that might underlie these effects. Based on these recent findings, we argue that the role of testosterone in human social behavior might be best understood in terms of the search for, and maintenance of, social status. PMID- 21616703 TI - Prospective multicenter trial assessing effectiveness, refractive predictability and safety of a new aberration free, bi-aspheric intraocular lens. AB - PURPOSE: To determine the effectiveness and safety of the Softec HD IOL; and to present refractive outcomes for lenses manufactured at an IOL power tolerance of 0.11 D. METHODS: Three-hundred and ninety adult patients requiring removal of a cataractous lens with implantation of a monofocal IOL in at least one eye were eligible for study participation across eight US investigative sites. Patients were enrolled unilaterally. After routine surgery, subjects were examined for adverse events (AEs), best corrected visual acuity (BCVA) and manifest refraction correction at 12 months postoperatively. RESULTS: Three-hundred and sixty-six (95%) of patients completed the 12-month postoperative visit. The percent of patients achieving best corrected Snellen acuity 20/40 or better was 98.9%, and 81.1% of patients achieved best corrected Snellen acuity 20/25 or better. Of those patients (80%) implanted with a lens available in 0.25 D increments (manufactured at a tolerance of 0.11 D) 40.9%, 69.8% and 93.8% of patients were within +/-0.25 D, +/-0.50 D and +/-1.0 D of predicted target refraction respectively. Overall incidence of cumulative and persistent IOL Grid AEs was 2.2% with no AE meeting or exceeding the FDA Grid of Historical Controls. CONCLUSIONS: The Softec HD IOL is a safe and effective lens. The high manufacturing tolerance of the lens appears to enhance refractive outcomes. PMID- 21616704 TI - Rapid green synthesis of gold nanoparticles using Rosa hybrida petal extract at room temperature. AB - This study reports a green method for the synthesis of gold nanoparticles using the aqueous extract of rose petals. The effects of gold salt concentration, extract concentration and extract quantity were investigated on nanoparticles synthesis. Gold nanoparticles were characterized with different techniques such as UV-vis spectroscopy, FT-IR spectroscopy, X-ray diffraction, energy dispersive X-ray spectroscopy, dynamic light scattering and transmission electron microscopy. Transmission electron microscopy experiments showed that these nanoparticles are formed with various shapes. FT-IR spectroscopy revealed that gold nanoparticles were functionalized with biomolecules that have primary amine group (-NH2), carbonyl group, -OH groups and other stabilizing functional groups. X-ray diffraction pattern showed high purity and face centered cubic structure of gold nanoparticles. Dynamic light scattering technique was used for particle size measurement, and it was found to be about 10nm. The rate of the reaction was high and it was completed within 5 min. PMID- 21616705 TI - On the infrared spectrum of L-lysinium(2+) sulfate. AB - It is argued that the infrared spectrum of L-lysine sulfate (L-Lys2+.SO4(2-)) published by Krishnakumar et al. [V. Krishnakumar, S. Manohar, R. Nagalakshmi, Spectrochim. Acta A75 (2010) 1394-1397] cannot belong to this crystal species. To confirm this, crystals (L-Lys2+.SO4(2-)) were synthesized and identified by XRD, and ATR FTIR and Raman spectra of L-Lys2+.SO4(2-) were registered and are discussed. PMID- 21616706 TI - Study on naringenin-CTMAB-DNA system by resonance light scattering technique and its analytical application. AB - A new high-sensitivity determination method of deoxyribonucleic acid (DNA) with detection limit at nanogram levels was proposed. Based on the measurement of resonance light scattering (RLS), it was found DNA could combine with naringenin and cetyltrimethylammonium bromide (CTMAB) in basic Tris-HCl buffer and produce enhanced RLS signal. The optimum conditions for this system were studied in detail. The enhanced intensity of RLS of naringenin-CTMAB at 353 nm was directly proportional to the concentration of DNA in the range of 0.017-1.7 MUg mL(-1). The detection limit was 5.06 ng mL(-1). Using the proposed method, the synthetic samples were analyzed with satisfactory results, the recovery was 99.3-105.0% and RSD was 0.7-3.7%. PMID- 21616707 TI - Molecular structure and infrared spectra of (HXeCN)n (n=2, 3 or 4). AB - The structure, energetics, and vibrational spectra of the (HXeCN)2 dimer were investigated at the CCSD(T), MP2 and B3LYP levels. Such properties of the (HXeCN)3 trimer and (HXeCN)4 tetramer were investigated at the B3LYP level. The dimer, trimer, and tetramer were predicted to have a C2h, C2v, and D2d structure, respectively. In all of these oligomers, the N?Xe intermonomeric interaction is the most important one for holding the monomers together. Included with the ZPVE and BSSE, the stabilization energy of the dimer is 12.36 kcal/mol at the CCSD(T) level, while those of the dimer, trimer, and tetramer are 10.42, 18.23, and 31.34 kcal/mol, respectively, at the B3LYP level. At the B3LYP level, with respect to those of the isolated monomer, the C-Xe and Xe-H asymmetric stretching frequencies are shifted by -11.2 and +128.0 cm(-1) for the dimer, -51.6, +220.7 and -11.5, +96.6 cm(-1) for the trimer, and -14.1 and +201.8 cm(-1) for the tetramer. PMID- 21616708 TI - Synthesis, molecular modeling, thermal and spectral studies of metal complexes of hydrazone derived from 5-acetyl-4-hydroxy-2H-1,3-thiazine-2,6(3H)-dione and thiosemicarbazide. AB - Metal complexes with the general formula [ML(H2O)(CH3OH)x].nH2O.(CH3OH)y(NO3)z [M=Cu(II), Ni(II), Co(II), VO(IV), Cr(III), Cd(II), Zn(II) or UO2(VI); x=0-2; y=0,1; z=0,1; n=0-2, 6 and L=hydrazone (H2L) derived from condensation of thiosemicarbazide with 5-acetyl-4-hydroxy-2H-1,3-thiazine-2,6(3H)-dione. The synthesized ligand and its metal complexes have been characterized on the basis of elemental analyses, spectral and magnetic studies as well as thermal gravimetric analysis (TGA). The deprotonated ligand acts as a dibasic tridentate (ONS) via phenolate oxygen, azomethine (CN), and thiolate (C-S) groups. Copper(II) complex exhibits square planar geometry. Nickel(II), chromium(III) and dioxouranium(VI) complexes exhibit octahedral geometry. Cobalt(II), cadmium(II) and zinc(II) complexes showed tetrahedral geometry, whereas oxovanadium(IV) reveals square pyramidal geometry. Thermal analysis are investigated and showed either three or four thermal decomposition steps. Kinetic parameters (Ea, A, DeltaH, DeltaS and DeltaG) of the thermal decomposition stages have been evaluated using Coats-Redfern equations. The molecular parameters of the ligand and its metal complexes have been calculated and correlated with the experimental data such as IR and TGA results. PMID- 21616709 TI - Incidental Lewy body disease: electrophysiological findings suggesting pre clinical Lewy body disorders. AB - OBJECTIVE: Evaluate electrophysiologic findings in incidental Lewy body disease (ILBD). METHODS: ILBD, Control, and Parkinson's disease (PD) subjects had electrophysiological evaluation within 2 years prior to autopsy. Data analyzed included surface electromyography (EMG) of upper extremity muscles during rest and muscle activation, and electroencephalography (EEG) recording at rest. For EMG, gross tracings and spectral peaks were analyzed. EEG measures analyzed were background frequency and power in delta, theta, alpha, and beta bands. RESULTS: Three of ten ILBD subjects (30%) showed unilateral rhythmic EMG discharges at rest without a visually apparent rest tremor. The ILBD resting EMG frequency was lower than in the Control group with no overlap (P=.03) and close to that of the PD group. The ILBD group had significantly lower background rhythm frequency than the Control group (P=.001) but was greater than the PD group (P=.01). CONCLUSIONS: The electrophysiologic changes in ILBD cases are between those of Control and PD, suggesting that these findings may reflect changes correlating with ILBD as a possible precursor to PD. SIGNIFICANCE: Electrophysiologic changes in ILBD may assist with the identification of a preclinical stage for Lewy body disorders and help the development of a therapeutic agent for modifying Lewy body disease progression. PMID- 21616710 TI - Assembly of a protein "brush" by end-grafting titin fragments to liposomes. AB - Titin is a giant filamentous protein of striated muscle composed of >300 immunoglobulin-like domains linked in tandem. Here, we demonstrate that a six immunoglobulin fragment of titin carrying a poly-histidine tag forms a protein "brush" on liposomes containing metallochelating lipids. These specimens might allow the direct visualization by cryo-EM of frozen hydrated and unstained titin chains with preserved architectural features. PMID- 21616712 TI - Structures and functions of autotransporter proteins in microbial pathogens. AB - Since their discovery more than 20 years ago the autotransporter protein superfamily has been growing continuously and currently represents the largest protein family in (pathogenic) Gram-negative bacteria. Autotransporter proteins (AT) adhere to a common structural principle and are composed of a C-terminal beta-barrel-shaped 'translocator' domain and an N-terminal 'passenger' domain. The translocator is anchored in the outer membrane and is indispensable for the N terminal passenger part to traverse the outer membrane. Most if not all AT harbor a chaperone segment that increases protein stability and may be located in the passenger or translocator domain. The passenger mediates the specific virulence function(s) of the particular AT. Accordingly, passenger domains of AT can be quite variable. Interestingly, AT have been identified as the first glycosylated proteins in Gram-negative bacteria. Despite the considerable efforts invested in the characterization of autotransporter biogenesis, various aspects such as the participation of accessory proteins, the fate of the translocator, or the translocation of glycosylated proteins still remain only poorly understood. In addition, recent evidence indicates that the prefix 'auto' might be slightly exaggerated. Here, we will selectively discuss novel insights at various stages of AT biogenesis. PMID- 21616711 TI - High prevalence of aggregative adherent Escherichia coli strains in the mucosa associated microbiota of patients with inflammatory bowel diseases. AB - The intestinal population of Escherichia coli is increased in patients with inflammatory bowel disease (IBD), but the reason for this elevation, the particular features of these bacteria and their potential role in the pathogenesis of the disease are not known. The present study was undertaken to investigate the adherence abilities and some virulence properties of a collection of 131 E. coli isolates cultured from rectal biopsies of 23 subjects diagnosed with ulcerative colitis (UC), 8 with Crohn's disease (CD) and 23 control patients from southern Brazil. The adherence abilities of the bacteria were investigated in vitro, using HEp-2 epithelial cells in assays of 3 and 6h of bacteria-cell contact. The isolates were screened by PCR with primers for the following virulence genetic markers: plasmid of aggregative adhesion (pAA) and the aggregative adherence fimbriae R (aggR), E. coli attaching and effacing (eae), invasion-associated locus (ial), invasion plasmid antigen H (ipaH) and Shiga citotoxin-encoding (stx) genes. HEp-2 cells aggregative adherent E. coli strains, as detected in the 3h adherence assay, were found in 14/23 (60.9%) patients with UC, 7/8 (87.5%) with CD and in 7/23 (30.4%) controls (p=0.011). Virulence genetic markers were detected in strains of 9 patients with UC (39.1%), but in none of CD or control group. Two of these UC patients had strains harboring both pAA and aggR, one had strains positive for aggR, four had strains positive for eae and two had strains positive for stx. These results suggest that the augmented population of E. coli on the rectal mucosa of IBD patients, particularly of those diagnosed with UC, is mostly comprised of aggregative adherent strains, some of which possessing classical virulence markers of E. coli. PMID- 21616713 TI - Pilot study on the exposure of the German general population to non-dioxin-like and dioxin-like PCBs. AB - In the past, polychlorinated biphenyls (PCBs) have been widely used and were distributed in the environment. Due to their high persistence and bioaccumulative potential, they can still be detected in the blood of the general population, despite their ban more than 20 years ago. Among the various congeners, the presence of dioxin-like PCBs in blood raises the highest environmental concerns due to their critical toxicological properties. We determined the plasma concentrations of 6 non-dioxin-like and 12 dioxin-like PCBs as well as the organochlorine pesticides HCB and p,p'-DDE (p,p' dichlorodiphenyldichloroethylene) in a group of 105 non-smokers out of the German general population by GC/MS as an estimate of the background burden to these compounds. The organochlorine compounds were extracted from plasma with n-hexane, cleaned up on a silica gel column and finally quantified using GC/MS and (13)C(12)-labelled internal standards. The limit of detection for all congeners was determined to be 0.01 MUg/Lplasma. Age had a strong influence on the plasma levels of HCB (hexachlorobenzene), p,p'-DDE and all higher chlorinated PCB congeners, while no gender difference was observed. Among the dioxin-like PCBs, PCB 118, PCB 156 and PCB 167 were detectable in 74-98% of all samples. Highly significant correlations between several PCB-congeners were observed. Our results provide for the first time volume-based data on the extent of the age-related background burden to dioxin-like PCBs in Germany. PMID- 21616714 TI - Validation and comparison of ActiGraph activity monitors. AB - OBJECTIVE: To compare activity counts from the ActiGraph GT3X to those from the ActiGraph GT1M during treadmill walking/running. A secondary aim was to develop tri-axial vector magnitude (VM3) cut-points to classify physical activity (PA) intensity. METHODS: Fifty participants wore the GT3X and the GT1M on the non dominant hip and exercised at 4 treadmill speeds (4.8, 6.4, 9.7, and 12 km h( 1)). Vertical (VT) and antero-posterior (AP) activity counts (counts min(-1)) as well as the vector magnitudes of the two axes (VM2) from both monitors were tested for significant differences using two-way ANOVA's. Bland-Altman plots were used to assess agreement between activity counts from the GT3X and GT1M. Linear regression analysis between VM3 countsmin(-1) and oxygen consumption data was conducted to develop VM3 cut-points for moderate, hard and very hard PA. RESULTS: There were no significant inter-monitor differences in VT activity counts at any speed. AP and VM2 activity counts from the GT1M were significantly higher (p<0.01) than those from the GT3X at 4.8, 9.7 and 12 km h(-1). High inter-monitor agreement was found for VT activity counts but not for AP and VM2 activity counts. VM3 cut-points for moderate, hard, and very hard PA intensities were 2690 6166, 6167-9642, >9642 counts min(-1). CONCLUSION: Due to the lack of congruence between the AP and VM2 activity counts from the GT1M and the GT3X, comparisons of data obtained with these two monitors should be avoided when using more than just the VT axis. VM3 cut-points may be used to classify PA in future studies. PMID- 21616715 TI - Symptoms and signs associated with syncope in young people with primary cardiac arrhythmias. AB - BACKGROUND: It is often reported that clinical symptoms are useful in differentiating cardiac from non-cardiac syncope. Studies in the young are rare. This study was designed to capture the symptoms and signs reported by patients with cardiac syncope before the patients or their attending clinicians knew the final diagnosis. METHODS: Retrospective case-note review of 35 consecutive unrelated gene-positive probands with a proven cardiac channelopathy. RESULTS: The presentation leading to diagnosis of cardiac channelopathy was resuscitated sudden cardiac death in 7 patients; syncope in 20; collapse with retained consciousness in 2; palpitations in 1 and an incidental finding in 5. For the 20 patients with syncope (LQTS 18, Brugada syndrome 2), median age at presentation was 13.9 years (1.8 day to 40.8 years). Of the 17 patients able to describe the onset of syncope, 11 (65%) had at least one symptom prior to collapse, though none reported nausea. Dizziness or lightheadedness was the most frequent symptom, being experienced by 8 (47%). Nine (of 20) patients (45%) had witnessed seizure like activity and 8 (40%) had urinary incontinence. Nineteen patients were capable of describing the post-syncopal period, of whom 15 (79%) reported symptoms, the most common (12; 65%) being drowsiness or exhaustion. CONCLUSIONS: Cardiac syncope in the young frequently presents with symptoms and signs that are typically associated with other causes of transient loss of consciousness, including vasovagal syncope and seizure disorders. The presence of symptoms may not be as helpful in differentiating arrhythmic from non-arrhythmic events as is often supposed. A thorough history, appropriate investigations and a high index of suspicion remain essential in the assessment of syncope. PMID- 21616716 TI - Self-perceived burden in patients with cancer: scale development and descriptive study. AB - PURPOSE: The purpose of this study was to develop a Japanese version of the Self Perceived Burden Scale (SPBS) for patients with cancer, to confirm its validity and reliability, and to elucidate the features of Self-Perceived Burden in Japanese patients with cancer. METHODS: Firstly, study team members carried out repeated translation and back translation. After that, scale development involved several phases including item development, construct validity testing, criterion related validity testing, and reliability testing. A questionnaire that included the SPBS was distributed to 310 patients with cancer. Responses were received from 226 (72.9%) patients and final analysis was performed on data from 210 of these. RESULTS: Exploratory factor analysis indicated that the 18-item version (alpha = 0.96) of the scale consisted of a single main factor. A 9-item abbreviation (alpha = 0.93) was therefore proposed. The convergent correlations of the SPBS with the FACIT-Sp and GHQ-12 scales showed significant correspondence. The construct of SPB was distinct from the "social and family well-being" FACIT-Sp subscale. Performance status, disease duration, living arrangement, and main caregiver were significantly related to the SPBS. CONCLUSION: We developed a Japanese version of the SPBS for cancer patients and demonstrated its validity and reliability, which makes it possible to study SPB in cancer patients in Japan. Our study revealed that cancer patients in Japan also feel SPB, in a way that relates specifically to their cultural background. PMID- 21616717 TI - Mucinous carcinomas of the ovary and colorectum: different organ, same dilemma. AB - Mucinous carcinomas are uncommon histological types that affect several organ sites. Primary mucinous carcinomas of the ovary are distinct from other ovarian carcinoma types, but they can pose a particular challenge for correct diagnosis from metastases, which most usually originate from the colorectum. Correct diagnosis is the mainstay of treatment, because standard practice states that protocols are tailored to the primary organ site. Little is known of mutational alterations in primary and metastatic mucinous carcinomas of the ovary, and few markers exist that can discriminate between them. We reviewed commonalities between ovarian and colorectal mucinous carcinomas with respect to aetiology, molecular alterations, differential diagnosis, and implications for treatment. Although primary mucinous carcinomas of the ovary and colorectum share similar mutational patterns and unfavourable outcomes at advanced stage, compared with their non-mucinous counterparts, important differences exist with respect to mucin localisation and specific molecular alterations. Technologies--eg, next generation sequencing--could aid identification of additional driver molecular changes that will help clarify the relation between mucinous carcinomas from different organ sites. Perhaps, then, we can consider moving towards testing and adoption of therapeutic approaches tailored to molecular characteristics of mucinous carcinomas, irrespective of organ site, so patients' survival can be optimised. PMID- 21616718 TI - Defining macrophage phenotype and function in adipose tissue. AB - In obesity, chronic low-grade inflammation is thought to mediate the effects of increased adipose tissue mass on metabolic comorbidity. Of the different cell types that contribute to obesity-induced inflammation in adipose tissue, this review focuses on macrophages and their monocytes precursors. Mechanisms for monocyte recruitment to adipose tissue, and how both monocytes and macrophages are phenotypically modified in this environment in response to increasing fat mass, are considered. The versatile phenotype of adipose tissue macrophages might contribute not only to inflammatory and metabolic alterations, but could also help to maintain adipose tissue homeostasis in the setting of obesity. PMID- 21616719 TI - Transition from open to robotic-assisted pediatric pyeloplasty: a feasibility and outcome study. AB - PURPOSE: Laparoscopic reconstructive procedures in the pediatric patient are associated with a steep learning curve. Outcomes from robotic-assisted pediatric urology have been reported by surgeons with known facility in laparoscopic surgery. We describe the experience of a single surgeon in transitioning from open to robotic-assisted laparoscopic pyeloplasty (RALP) without previous training in traditional laparoscopic pyeloplasty or intracorporeal suturing. MATERIALS AND METHODS: We reviewed our experience with 20 (mean age 7.4 years) consecutive children undergoing RALP for ureteropelvic junction obstruction at our institution over 36 months. Additionally, a literature search was conducted to identify age-similar patient groups who underwent open and laparoscopic pyeloplasty. RESULTS: Length of hospitalization and postoperative analgesia requirement were greater in the age-similar open pyeloplasty group compared to the other two groups. Intraoperative times were greater in the laparoscopic and RALP groups compared to the open pyeloplasty group. CONCLUSIONS: Our experience confirms the feasibility of transitioning from open to robotic-assisted laparoscopic pediatric pyeloplasty without previous experience in conventional laparoscopy. Outcomes, analgesic requirement and hospitalization for the patients from our institution are comparable to the laparoscopy patient group and improved compared to open pyeloplasty patients from the literature. PMID- 21616720 TI - Smeulders N, Makin E, Desai D, Duffy PG, Healy C, Cuckow PM et al. The predictive value of a repeat micturating cystourethrogram for remnant leaflets after primary endoscopic ablation of posterior urethral valves. J Pediatr Urol 2011; 7: 203-8. PMID- 21616721 TI - Rural and urban differences in the associations between characteristics of the community food environment and fruit and vegetable intake. AB - OBJECTIVE: To examine the relationship between measures of the household and retail food environments and fruit and vegetable (FV) intake in both urban and rural environmental contexts. DESIGN: A cross-sectional design was used. Data for FV intake and other characteristics were collected via survey instrument and geocoded to the objective food environment based on a ground-truthed (windshield audit) survey of the retail food environment. SETTING: One urban and 6 contiguous rural counties. PARTICIPANTS: This study involved 2,556 residents of the Brazos Valley, Texas, who were selected through random-digit dialing. MAIN OUTCOME MEASURE: Two-item scale of FV intake. ANALYSIS: Data were analyzed using chi square analysis, 2-sample t tests, and linear regression. RESULTS: Distance to supermarket or supercenter was insignificant in the urban model, but significant in the rural model (beta = -.014, P < .010, confidence interval = -.024, -.003). CONCLUSIONS AND IMPLICATIONS: Retail food environments have different impacts on FV intake in urban and rural settings. Interventions to improve FV intake in these settings should account for the importance of distance to the retail food environment in rural settings. PMID- 21616722 TI - Out-of-pocket drug costs and drug utilization patterns of postmenopausal Medicare beneficiaries with osteoporosis. AB - BACKGROUND: The Medicare Part D coverage gap has been associated with lower adherence and drug utilization and higher discontinuation. Because osteoporosis has a relatively high prevalence among Medicare-eligible postmenopausal women, we examined changes in utilization of osteoporosis medications during this coverage gap. OBJECTIVES: The purpose of this study was to investigate changes in out-of pocket (OOP) drug costs and utilization associated with the Medicare Part D coverage gap among postmenopausal beneficiaries with osteoporosis. METHODS: This retrospective analysis of 2007 pharmacy claims focuses on postmenopausal female Medicare beneficiaries enrolled in full-, partial-, or no-gap exposure standard or Medicare Advantage prescription drug plans (PDPs), retiree drug subsidy (RDS) plans, or the low-income subsidy program. We compared beneficiaries with osteoporosis who were taking teriparatide (Eli Lilly and Company, Indianapolis, Indiana) (n = 5657) with matched samples of beneficiaries who were taking nonteriparatide osteoporosis medications (NTO; n = 16,971) or who had other chronic conditions (OCC; n = 16,971). We measured average monthly prescription drug fills and OOP costs, medication discontinuation, and skipping. RESULTS: More than half the sample reached the coverage gap; OOP costs then rose for teriparatide users enrolled in partial- or full-gap exposure plans (increase of 121% and 186%; $300 and $349) but fell for those in no-gap exposure PDPs or RDS plans (decrease of 49% and 30%; $131 and $40). OOP costs for beneficiaries in partial- or full-gap exposure PDPs increased >120% (increase of $144 and $176) in the NTO group and nearly doubled for the OCC group (increase of $124 and $151); these OOP costs were substantially lower than those for teriparatide users. Both teriparatide users and NTO group members discontinued or skipped medications more often than persons in the OCC group, regardless of plan or benefit design. CONCLUSION: Medication discontinuation and OOP costs among beneficiaries with osteoporosis were highest for those enrolled in Part D plans with a coverage gap. Providers should be aware of potential cost-related nonadherence among Medicare beneficiaries taking osteoporosis medications. PMID- 21616723 TI - Comment on: Reduction in anastomotic strictures using bioabsorbable circular staple line reinforcement in laparoscopic gastric bypass. PMID- 21616724 TI - Practice of routine intraoperative leak test during laparoscopic sleeve gastrectomy should not be discarded. PMID- 21616725 TI - Management of gastrointestinal leaks after surgery for clinically severe obesity. AB - BACKGROUND: Gastrointestinal leaks after bariatric surgery are the primary cause of serious morbidity and mortality nationwide. Enteric leaks can differ in severity, presentation, and management, depending on the type of bariatric surgery performed. Our objective was to describe the clinical presentation and treatment outcomes in patients who developed postoperative leaks at a university hospital bariatric referral center. METHODS: A retrospective observational study using descriptive statistics was conducted on data from 1499 bariatric operations performed at our institution from 1994 to 2010. The procedures included a variant of biliopancreatic diversion with long limb reconstruction (BPD-LL) in 820 patients (791 open and 29 laparoscopic), Roux-en-Y gastric bypass (RYGB) in 301 patients (105 open and 196 laparoscopic), and sleeve gastrectomy (SG) in 208 patients (5 open and 203 laparoscopic). RESULTS: Of these patients, 30 (2%) developed a postoperative leak at a median of 18 days (range 2-32) postoperatively. The primary procedure was laparoscopic SG in 12 patients (5.8%), laparoscopic RYGB in 5 patients (1.6%), and BPD-LL (12 open and 1 laparoscopic) in 13 patients (1.6%). In all patients who underwent laparoscopic SG, the leak site was along the staple line. The gastrojejunal anastomosis was leaking in 4 (80%) and 12 (92.3%) patients in the RYGB and BPD-LL group, respectively. The enteroenteral anastomosis was leaking in 1 patient each in the RYGB and BPD-LL groups (20% and 7.7%, respectively). Three patients (10%; 2 from the BPD-LL group and 1 from the RYGB group) presented with generalized peritonitis and underwent emergency re-exploration; nonoperative treatment was successful in the remaining 27 patients (90%). Stent placement for persistent gastrocutaneous fistula was used in 9 patients (30%; 8 from the SG cohort and 1 from the BPD-LL group). The overall mortality rate was 3.3%. CONCLUSION: In our experience, most leaks resulting from antiobesity surgery were successfully managed using nonoperative methods. Rapid management of gastrointestinal leaks using computed tomography guided drainage and/or intraluminal stent placement could be the treatment of choice in selected patients. PMID- 21616726 TI - Embolic capture angioplasty of lower extremity lesion following distal embolization. AB - Peripheral embolization is a known complication of peripheral artery interventions. It can lead to occlusion of distal vessels leading to significant lower extremity ischemia and complications. Peripheral artery interventions involving chronic total occlusions have been shown to have higher rates of complications including distal embolization. Although distal embolic protection strategies are available, they are seldom utilized during lower extremity interventions, especially during treatment of totally occluded vessels. Proteus embolic capture angioplasty balloon may provide operators with the option of balloon dilation of stenotic lesion in the peripheral arterial bed, along with embolic capture. We report a case where use of this novel device helped in achieving a successful angioplasty outcome along with evacuation of distal embolus and restoration of antegrade flow. PMID- 21616728 TI - An improved approach for extraction and high-performance liquid chromatography analysis of paraquat in human plasma. AB - A simple, sensitive, reliable and economical HPLC method for quantifying paraquat concentration in human plasma has been developed, using diethyl paraquat as an internal standard. The drugs were extracted from the sample and separated on Xtimate C18 column with a mobile phase of 15% acetonitrile in 0.1M orthophosphoric acid containing SDS (150 mg/l). The pH of the mobile phase was adjusted to 3 with triethylamine and the detection wavelength was 256 nm for both paraquat and the internal standard. The average extraction recoveries were 91.9%. Good linearity (R(2)=0.9984) was observed throughout the range of 0.02-10 MUg/ml in 0.5 ml plasma. The overall accuracy of this method was 97.6-107.3% and the lower limit of detection was 0.01 MUg/ml. The intra- and inter-day variations were lower than 3.65% and 2.64%, respectively. We used this method to examine the paraquat concentrations of 53 patients with acute paraquat intoxication of whom 26 (49.1%) survived. In conclusion, this method was suitable for quantification of paraquat plasma concentration in toxicological samples. It was helpful in both assessing the severity of intoxication and predicting the outcome of paraquat poisoning. PMID- 21616727 TI - Intramural coronary lipid injection induces atheromatous lesions expressing proinflammatory chemokines: implications for the development of a porcine model of atherosclerosis. AB - BACKGROUND: Intramural delivery of lipids into the coronaries of pigs fed high cholesterol diet results in the formation of localized atherosclerotic-like lesions within 12 weeks. These lesions are located in positively remodeled vessels and are associated to the development of abundant adventitial vasa vasorum and mononuclear cell infiltrate. In this study, we aimed to analyze the degree of expression of various inflammatory chemokines within the developed lesions compared with control segments injected with saline. METHODS: Balloon injury was performed in 15 coronary arteries of pigs fed high-cholesterol diet for 12 weeks. Two weeks after procedure, 60 coronary segments were randomized to either intramural injections of complex lipids (n=30) or normal saline (n=30). Neovessel density in the lesions was analyzed by lectin stain. Segments were processed for RNA expression of inflammatory chemokines such as monocyte chemoattractant protein-1 and vascular endothelial growth factor. RESULTS: At 12 weeks, the percentage area of stenosis seen in histological sections was modest in both groups (lipids: 17.3+/-15 vs. saline: 32.4+/-22.8, P=.017). The lipid group showed higher vasa vasorum (VV) quantity (saline: 18.2+/-14.9 VV/section vs. lipids: 30.6+/-21.6 VV/section, P<.05) and vasa vasorum density (saline: 7.3+/-4.6 VV/mm(2) vs. lipids: 16.5+/-9 VV/mm(2), P<.001). In addition, monocyte chemoattractant protein-1 expression was higher in the lipid group (1.5+/-1.12) compared with saline control group (0.83+/-0.34, P<.01). Vascular endothelial growth factor expression was also higher in the lipid group (1.36+/-0.9) compared with saline group (0.87+/-0.33, P<.05). CONCLUSION: The intramural injection of complex lipids into the coronary arteries of pigs maintained in a high cholesterol diet results in focal lesions located in positively remodeled vessels that have a high neovessel count and express proinflammatory chemokines. PMID- 21616729 TI - Why does height matter for educational attainment? Evidence from German children. AB - Height is positively associated with educational attainment. We investigate the mechanisms behind this relationship using data on German pre-teen students. We show that taller children are more likely to enroll in Gymnasium, the most academic secondary school track, and that primary school teachers provide more favorable school track decisions to taller students. We find that a 1cm increase in height is associated with a 1.6 percentage points increase in the probability of attending Gymnasium. This holds even when controlling for academic achievement and parental background. In addition, we present evidence that height and social skills are positively associated already at age 2-3. We propose the association between height and noncognitive skills as a possible explanation of the height school premium, even if discrimination cannot be ruled out entirely. PMID- 21616730 TI - Multiple varieties of musical meaning: Comment on "Towards a neural basis of processing musical semantics" by Stefan Koelsch. PMID- 21616731 TI - Gastric metastasis of breast cancer: a single centre retrospective study. AB - BACKGROUND: Digestive metastasis of breast cancer are rare but when they do occur the stomach is one of the commoner sites. AIM: To describe the clinical, endoscopic, pathological features and treatment. METHODS: 35 cases of gastric metastasis were identified retrospectively between 1980 and 2008. RESULTS: The location of the gastric metastasis was fundus (n=15, 43%), antrum (n=15, 43%) or both (n=5, 14%). The histological subtype of primary breast cancer was invasive lobular carcinoma in 34 patients (97%). Hormonal receptors were positive in 19 out of 24 cases (79%), two out of 22 analysed were HER2 positive (9%). There were 16 (46%) patients with peritoneal carcinosis. The treatment was chemotherapy (n=13, 37%), hormonotherapy (n=2, 6%) or both (n=13, 37%). The 2-year survival rate after gastric metastasis diagnosis was 53% with a median follow up of 31 months [7-84 months]. CONCLUSION: Ninety-seven percent of gastric metastasis from breast cancers are derived from invasive lobular carcinoma. Seventy-nine percent of these are HER+ and comparison with the original histopathological slides of primary breast carcinoma should be performed to differentiate gastric metastasis from primary gastric carcinoma. Peritoneal carcinomatosis accompanied gastric metastasis in almost half the cases in this series and treatment was generally chemotherapy. PMID- 21616732 TI - Diabetes mellitus and the risk of cholecystectomy. AB - BACKGROUND: Whether diabetes mellitus is associated with an increased risk of cholecystectomy remains controversial. AIMS: To explore the association between diabetes mellitus and the risk of cholecystectomy. METHODS: Population-based case control analysis using UK-based General Practice Research Database. Cases of cholecystectomy and up to four controls per case, matched on age, sex, BMI, general practice, calendar time, and years of history in the database were identified between 1994 and 2008. Conditional logistic regression was used to estimate the risk of cholecystectomy in diabetics compared to non-diabetics. Odds ratios (ORs) were calculated, adjusted for smoking, alcohol consumption, statin use, and additional confounders. RESULTS: Amongst 22,574 cases with cholecystectomy and 72,476 controls, 1068 (4.7%) and 3270 (4.5%) had diabetes, respectively, yielding an adjusted OR for developing gallstone disease followed by cholecystectomy of 0.88, 95% CI 78-1.00, p=0.05) in association with diabetes mellitus. Neither glycaemic control, nor increasing diabetes duration or oral antidiabetic therapies were associated with an altered risk of cholecystectomy. Use of statins was protective in patients with (adj. OR 0.66, 95% CI 0.54-0.80, p<0.0001) or without diabetes (adj. OR 0.70, 95% CI 0.62-0.78, p<0.0001). CONCLUSIONS: Diabetes mellitus was not associated with an altered risk of cholecystectomy. PMID- 21616734 TI - Invited commentary. PMID- 21616733 TI - Management of chronic liver disease by general practitioners in southern Italy: unmet educational needs. AB - BACKGROUND: Despite chronic liver diseases represent an important cause of illness in Italy, data from family practice are poor. AIM: To assess the management of chronic liver diseases by general practitioners in a large area of Southern Italy. METHODS: This was a 5-year retrospective analysis from 104 physicians in charge of a population of 143,159 adult subjects. RESULTS: Amongst 6550 patients with chronic liver disease (4.7%, 3400 M, median age 57 years), 1330 (20.3%) had HCV infection, 226 (3.4%) HBV infection, and 293 (4.5%) liver cirrhosis (25 alcohol-related). The prevalence of alcohol consumption, recorded by 90% of physicians, was 20.4%. Hypertransaminasemia and liver steatosis had a prevalence of 6.7% and 2.4%, respectively. Although transaminases were checked 3 times over 5 years in 80% of cases, few patients were investigated for viral infection, and less than 50% underwent ultrasonography and consultation, leaving undefined a consistent number of cases. Alcohol consumption, body mass index and ultrasonography were poorly checked even in hypertransaminasemic patients. CONCLUSIONS: This study shows that data recording by general practitioners in chronic liver disease patients lacks homogeneity and can miss important information. One unmet need is therefore the integration between theoretical knowledge and practice to share similar behaviours and improve the management of these patients. PMID- 21616735 TI - Long-term results after 40 years experience with treatment of rare facial clefts: Part 2--Symmetrical median clefts. AB - BACKGROUND: Median facial clefts are reconstructive challenges, requiring multiple operations throughout life. Long-term results are often still far from ideal and could be improved. Due to surgical intervention and diminished intrinsic growth potential, surgical results may change from initially good into a progressively disappointing outcome. If, however, the ideal timing and type of surgery are known, in combination with the intrinsic growth potential, the results can be ameliorated. A guideline for surgical treatment is given. METHODS: Twenty patients with a pure symmetrical median cleft were evaluated on intermediate and long-term surgical results. The final result was scored based on severity of the initial and the remaining facial deformities, and the need for revisional surgery. RESULTS: The long-term surgical outcome was initially good for each of the affected facial parts and the face in general, but worsened over time, especially in the zone of the nose. An adequate and stable result of hypertelorism correction was observed for both the orbital box osteotomy and medial faciotomy, even when performed at a young age. CONCLUSIONS: The intrinsic growth restriction is mainly localised in the central midface. This leads to a complex and often unpredictable growth of the maturing face. It makes it difficult to achieve perfect reconstructions. Caution with surgical interventions of the nose at a young age is required. Once the face has matured, a midface advancement and secondary nose correction should be considered for satisfactory projection. Early referral to a specialised centre is essential. PMID- 21616736 TI - Bilateral trigger finger in a 5-year-old child: case report. AB - Trigger finger is rare in children. Herein, we report on a 5-year-old boy with trigger digits in several of his fingers since 1 month before which got worse. Triggering occurred in decreasing order of severity in proximal interphalangeal joints of the left middle, index, ring fingers and thumb. There was also triggering of the right index finger. The patient had no history of trauma, metabolic disorders and viral infections. The examination revealed triggering of the above-mentioned digits and was otherwise unremarkable. The patient was treated with physiotherapy (eight sessions) and non-steroidal anti-inflammatory drugs (NSAIDs) (for 20 days). The symptoms were completely resolved 4 months after its onset. Although surgical release is recommended by many authors for the treatment of trigger finger in children, if an inflammatory process is not unlikely, particularly when the symptoms present at older ages, the patient may be treated with a short course of empirical treatment with physiotherapy and/or NSAIDs to see his/her response. PMID- 21616737 TI - A model of translational research for diabetes prevention in low and middle income countries: The Diabetes Community Lifestyle Improvement Program (D-CLIP) trial. AB - AIMS: The Diabetes Community Lifestyle Improvement Program (D-CLIP) aims to implement and evaluate in a controlled, randomized trial the effectiveness, cost effectiveness, and sustainability of a culturally appropriate, low-cost, and sustainable lifestyle intervention for the prevention of type 2 diabetes mellitus in India. METHODS: D-CLIP, a translational research project adapted from the methods and curriculum developed and tested for efficacy in the Diabetes Prevention Program, utilizes innovated methods (a step-wise model of diabetes prevention with lifestyle and metformin added when needed; inclusion of individuals with isolated glucose tolerance, impaired fasting glucose, and both; classes team-taught by professionals and trained community educators) with the goals of increasing diabetes prevention, community acceptability, and long-term dissemination and sustainability of the program. The study outcomes are: diabetes incidence (primary measure of effectiveness), cost-effectiveness, changes in anthropometric measures, plasma lipids, blood pressure, blood glucose, and HbA(1c,) Program acceptability and sustainability will be assessed using a mixed methods approach. CONCLUSION: D-CLIP, a low-cost, community-based, research program, addresses the key components of translational research and can be used as a model for prevention of chronic diseases in other low and middle-income country settings. PMID- 21616738 TI - The evaluation and management of childhood type 2 diabetes mellitus. AB - Diabetes in children is increasing in prevalence with obesity-associated insulin resistance being a major contributing factor. This report contains a comprehensive review of treatment paradigms regarding the management of diabetes in children. The information provided was collected from an extensive literature review on type 2 diabetes, and children and/or adolescents with diabetes and/or obesity treatment. There is a wide array of contributing factors to the development of diabetes and consequences that indicate the need to tailor treatment plans with reducing obesity as a primary outcome. Lipocentric treatment paradigms offer opportunity to reduce the impact of obesity. PMID- 21616739 TI - Did Van Gogh have Meniere's disease? AB - Very surprising diagnoses have sometimes been made of the illness from which Vincent van Gogh suffered. Most of them can be safely ignored; the diagnosis of Meniere's disease, however, put forward by K. I. Arenberg, deserves attention. This diagnosis was first criticized in a doctoral thesis in 1992 in the university of Saint-Etienne (France), and again in a book published by Henri Andre Martin in 1994, and the present study is based on these. Analyzing van Gogh's pathology presupposes awareness of the family context in which he lived, his eventful life, his artistic career and of his correspondence, which notably provides no support for a diagnosis of Meniere's disease. Analysis, indeed, enables Meniere's disease to be categorically eliminated in favor of temporal epilepsy, as confirmed in 1956 by Gastaut following the diagnosis initially made half a century earlier by Dr Rey. Van Gogh's illness is an example of the contradictory changes in affect, behavior and personality to be observed in epileptic patients. Absinth intoxication doubtless aggravated van Gogh's epilepsy and may account for certain prolonged episodes of confusion. It would be wrong, however, to speak of alcoholic dementia rather than of a probable epileptic psychosis or other pathology such as hallucinatory psychosis, or delirious attack, either isolated or related to late-onset schizophrenia. Apart from certain episodes of severe mental disorder, however, van Gogh's exceptional creativity was maintained up to the very end. Like any other artist, his temperament was expressed in his art, exalted rather than impaired by his illness. PMID- 21616740 TI - Adenocarcinoma of the ethmoid sinus: retrospective study of 42 cases. AB - OBJECTIVES: Retrospective analysis of the oncological results and morbidity of ethmoid sinus adenocarcinoma surgery, and identification of survival factors. MATERIAL AND METHODS: Forty-two patients were treated from 1990 to 2009. The study covered clinical presentation, medical imaging, histologic data, TNM grade, treatment, morbidity and overall recurrence-free survival. RESULTS: Forty-one men and one woman, with mean age at diagnosis of 61.5 years, were included. 85.7% had been exposed to wood dust. Twenty patients (47.6%) were graded T4 at diagnosis. Thirty-three (78.5%) were treated by surgery followed by radiation therapy; nearly half of these showed recurrence. Overall specific 5-year survival was 44.2% at 5 years and recurrence-free 5-year survival 46.4%. The factors of poor prognosis found were cerebromeningeal or orbital invasion and local recurrence. CONCLUSION: This series confirms the epidemiological literature on ethmoid adenocarcinoma and the influence of poor prognosis factors. Survival depended directly on local tumor control and seemed to be enhanced on an isolated transfacial approach. PMID- 21616741 TI - Imaging IPMN: take home messages and news. AB - IPMN is a frequent disease involving pancreatic duct. This disease could be malignant (parenchymal invasive adenocarcinoma), particularly if the main pancreatic duct is involved (this involvement is considered present if > 6 mm), if this enlargement reaches 10 mm or more, and if the pathological phenotype is biliopancreatic or intestinal (malignancy is less frequent if gastric one). Invasiveness is suspected if hypodense parenchymal lesion is present, particularly near a cystical lesion or MPD, a mural nodule of the wall, or if MPD wall has got a contrast uptake. Mural nodules inside cystic branch duct are associated with in situ grade 3 malignancies. MPD IPMN must be resected to prevent malignancy. The follow-up of isolated branch duct cysts relies upon MDCT and MRI, every two years if lesion is less than 1cm. Every one year if bigger, particularly if more than to 3 cm. PMID- 21616748 TI - Chronic spontaneous urticaria is not an allergic disease. AB - The links between chronic urticaria, IgE sensitization and allergy have been much discussed but little studied. We investigated IgE sensitization and allergy in 128 adult chronic urticaria patients during 2006-2008. During a one-day hospitalisation, the patients answered a standardized questionnaire and underwent blood serum analysis, physical tests and skin prick-tests. IgE sensitization to environmental allergens was defined by the positivity of at least one skin prick test and/or elevated levels of serum IgE >= 300 Kui/L. The chronic urticaria was considered allergic if: i) a high correlation between positive skin prick tests to a clinically relevant allergen and the case history was found; ii) complete remission of urticaria occurred within two months of allergen withdrawal. Of 105 patients with interpretable skin prick tests, 46.7% were IgE sensitized. Two patients had clinically relevant positive skin prick tests but their chronic urticaria had many other triggering factors and neither was in complete remission after withdrawal of these allergens. IgE sensitization is higher in chronic urticaria patients than in the global adult population, suggesting that it is one important etiopathogenic factor in chronic urticaria. However, it cannot be considered as the expression of an IgE-mediated allergy but as a chronic inflammatory disease, more frequent in IgE sensitized people and favoured by multiple factors, among which IgE-mediated allergy is exceptional. PMID- 21616749 TI - The immune microenvironments of lung and intraocular tumors. AB - An increasing body of evidence underlines the prominent role of the immune microenvironment in cancer growth, invasion and metastasis. The local immune responses are specialized in the different organs. We will discuss the composition of the immune microenvironments and their role on tumor development in two cancers developing in opposite contexts: in the lung, which is at the interface of the outside world and in the eye, which is, an immunoprivileged site protected from it. We demonstrate that the immune system plays a major role in control of tumors, despite of these highly different tissue microenvironments. PMID- 21616751 TI - Disseminated superficial actinic porokeratosis associated with pseudoxanthoma elasticum. PMID- 21616750 TI - Lichen scrofulosorum caused by pulmonary Mycobacterium avium complex (MAC) infection. PMID- 21616752 TI - CD10 expressed by fibroblasts and melanoma cells degrades endothelin-1 secreted by human keratinocytes. AB - Endothelin-1 (ET-1) is a potent multifunctional peptide linked to wound healing, pigmentation, carcinogenesis, and fibrosclerotic processes in the skin. Whereas ET-1 was thought to be digested by receptor-mediated endocytosis, it is also reported to be biochemically degraded by the neutral endopeptidase CD10 using kidney homogenates. Although keratinocytes (KC) and fibroblasts (Fb) are sources of both ET-1 and CD10, respectively, there is no report investigating the direct association between CD10 expression and its function in relation to ET-1 degradation in the skin. CD10 expression in melanoma cells is associated with clinical prognosis, suggesting an important role in the invasive and metastatic potential of melanoma cells. Here, cultured KC produced much higher amounts of ET 1 than did cultured Fb or melanoma cells. In contrast, KC and A375 melanoma cells did not express CD10, while Fb, SK-MEL-28 and G361 melanoma cells constitutively expressed CD10. KC-derived ET-1 was down-modulated by both CD10-positive Fb and CD10-positive melanoma cells, and the inhibition was partially reversed under substitution conditions using CD10-knockdown Fb or CD10-knockdown melanoma cells. This indicates that CD10 on cultured Fb and melanoma cells is biochemically active in the degradation or down-modulation of ET-1 secreted from KC. These findings may lead to better understanding of skin homeostasis and of the malignant potential of melanoma. PMID- 21616753 TI - Effectiveness of electrochemotherapy in treatment of a recurrent squamous cell carcinoma of the scalp. PMID- 21616754 TI - Nevus anemicus of the cheek with ipsilateral melanosis bulbi: an unusual example of didymosis. PMID- 21616755 TI - The role of suicide risk in the decision for psychiatric hospitalization after a suicide attempt. AB - BACKGROUND: Suicide prevention can be improved by knowing which variables physicians take into account when considering hospitalization or discharge of patients who have attempted suicide. AIMS: To test whether suicide risk is an adequate explanatory variable for predicting admission to a psychiatric unit after a suicide attempt. METHODS: Analyses of 840 clinical records of patients who had attempted suicide (66.3% women) at four public general hospitals in Madrid (Spain). RESULTS: 180 (21.4%) patients were admitted to psychiatric units. Logistic regression analyses showed that explanatory variables predicting admission were: male gender; previous psychiatric hospitalization; psychiatric disorder; not having a substance-related disorder; use of a lethal method; delay until discovery of more than one hour; previous attempts; suicidal ideation; high suicidal planning; and lack of verbalization of adequate criticism of the attempt. CONCLUSIONS: Suicide risk appears to be an adequate explanatory variable for predicting the decision to admit a patient to a psychiatric ward after a suicide attempt, although the introduction of other variables improves the model. These results provide additional information regarding factors involved in everyday medical practice in emergency settings. PMID- 21616756 TI - Specific psychological treatment versus treatment as usual in adolescents with self-harm: systematic review and meta-analysis. AB - BACKGROUND: Despite recent advances in the understanding and treatment of self harm, poor engagement with therapy remains a serious problem. AIMS: To investigate whether offering specific psychological treatment (SPT) leads to better engagement than offering treatment as usual (TAU) in adolescents who have self-harmed. METHODS: Data sources were identified by searching Medline, PsychINFO, EMBASE, and PubMed for randomized controlled trials comparing SPT versus TAU in adolescents presenting with self-harm. RESULTS: Seven studies met inclusion criteria, and six were entered into the meta-analysis. There was no statistically significant difference between the number of subjects not completing four or more sessions of an SPT (27.7%, 70/253) than TAU (43.3%, 106/245), RR = 0.71 (95% CI: 0.49-1.05). CONCLUSIONS: Engaging adolescents with psychological treatment is necessary although not sufficient to achieve treatment goals. Further research is needed to develop tools for maximizing engagement. PMID- 21616757 TI - Key considerations for preventing suicide in older adults: consensus opinions of an expert panel. AB - BACKGROUND: The number of older adults is growing rapidly. This fact, combined with the high rates of suicide in later life, indicates that many more older adults will die by their own hands before rigorous trials can be conducted to fully understand the best approaches to prevent late life suicide. AIMS: To disseminate key considerations for interventions addressing senior suicidal behavior. METHODS: An international expert panel has reviewed and discussed key considerations for interventions against suicide in older adults based on existing evidence, where available, and expert opinion. RESULTS: A set of new key considerations is divided into: universal, selective, and indicated prevention as well as a section on general considerations. CONCLUSIONS: The suggestions span a wide range and are offered for consideration by local groups preparing new interventions, as well as large scale public health care planning. PMID- 21616758 TI - Suicidal behavior on Axis VI: clinical data supporting a sixth Axis for DSM-V. AB - BACKGROUND: Oquendo and colleagues (Oquendo, Baca-Garcia, Mann, & Giner, 2008; Oquendo & Currier, 2009) recommend that DSM-V emphasize suicide risk assessment on a sixth axis, thereby increasing regularity of suicide risk assessments. AIMS: We propose that evidence of nonredundancy with Axis V - Global Assessment of Functioning (GAF) is one piece of data that can serve as a starting point for a line of research establishing incremental predictive utility for a separate suicide risk assessment in the DSM framework. METHODS: A standardized suicide risk assessment protocol, measures of depressive, anxious, and eating disordered symptomatology, as well as an index of comorbidity were administered to a sample of 412 adult outpatients. RESULTS: Our data indicate that data from standardized suicide risk assessments are associated with indices of symptomatology severity as well as comorbidity, controlling for GAF. CONCLUSIONS: These results support the nonredundancy of the assessments and suggest the utility of longitudinal investigations of the predictive utility of a sixth DSM axis in the assessment of suicide risk. PMID- 21616759 TI - Towards evidence-based suicide prevention programs. PMID- 21616760 TI - Self-report of suicidal thoughts and behavior vs. school nurse evaluations in Dutch high-school students. AB - BACKGROUND: Systematic screening to identify adolescents at risk for suicidal behavior is crucial to suicide prevention. AIMS: The current practice of screening and follow-up actions taken by school nurses in schools for secondary education in Rotterdam, the Netherlands, was studied. METHODS: In Rotterdam, an approach was developed in which, as a standard routine, students are invited to an interview with a school nurse after filling out a youth health survey in classroom setting. This health survey comprised, among other subjects, the Strengths and Difficulties Questionnaire (SDQ) and questions about suicidal thoughts and behaviors. The results from the interviews and the self-report questionnaires of nearly 4000 first-grade students in secondary education were analyzed. RESULTS: The vast majority of invitations were not followed by any action from the school nurse. Also, the majority of students whose self-report led to an "Attention Subject" for the nurse were not considered to be at risk by the nurse. The self-report of recent suicidal thoughts added to the predictive value of emotional distress as measured by the SDQ and other self-report conclusions. CONCLUSIONS: This study demonstrates the additional value of self reported suicidal thoughts for Dutch preventive practices in youth health care. PMID- 21616761 TI - The attitudes of preclinical and clinical Turkish medical students toward suicide attempters. AB - BACKGROUND: Interest in studies of stigma toward patients with mental disorders is growing. Research on the attitudes of medical students toward suicide attempters does not exist; although as medical personnel, they will encounter suicide attempters in emergency rooms. AIMS: We aimed to investigate the attitudes of preclinical and clinical medical students toward suicide attempters and to compare their attitudes with nonmedical students. METHODS: Participants were asked to fill out questionnaires those were searching the attitudes toward suicide attempters. These questionnaires used a social distance scale, skillfulness assessment scale, and dangerousness scale. RESULTS: More than 73% of students had a negative attitude toward "renting a room of their home to a suicide attempter." More than 90% would not want "a suicide attempter to supervise their children for few hours." Significantly more preclinical than clinical students would not want their children to marry a suicide attempter. CONCLUSIONS: Social distance, skillfulness, and attitudes concerning the dangerousness of suicide attempters are problematic for medical students and need educational intervention. PMID- 21616762 TI - Birthday blues: examining the association between birthday and suicide in a national sample. AB - BACKGROUND: Socioculturally meaningful events have been shown to influence the timing of suicide, but the influence of psychiatric disorder on these associations has seldom been studied. AIMS: To investigate the association between birthday and increased risk of suicide in the general population and in a national sample of psychiatric patients. METHODS: Data on general population suicides and suicide by individuals in recent care of mental health services were examined for day of death in relation to one's birthday using Poisson regression analysis. RESULTS: An increased risk of suicide was observed on day of one's birthday itself for males in both the general population (IRR = 1.39, 95% CI = 1.18-1.64, p < .01) and the clinical population (IRR = 1.48, 95% CI = 1.07-2.07, p = .03), especially for those aged 35 years and older. In the clinical population, risk was restricted to male patients aged 35-54 and risk extended to the 3 days prior to one's birthday. CONCLUSIONS: Birthdays are periods of increased risk for men aged 35 and older in the general population and in those receiving mental health care. Raising health-care professionals' awareness of patient groups at greater risk at this personally significant time may benefit care planning and could facilitate suicide prevention in these individuals. PMID- 21616763 TI - Hyperlinked suicide: assessing the prominence and accessibility of suicide websites. AB - BACKGROUND: The relationship between the Internet and suicide is a topic of growing concern among suicide researchers and the public, though to date few have actually attempted to investigate the accessibility and prominence of suicide related information online, and there have been no comprehensive studies of site networking structure. AIMS: To assess the visibility of various types of online information to suicide-risk individuals, and to assess the prominence and accessibility of "pro-suicide," suicide prevention, and support sites by measuring their networking structure. METHODS: Employing empirically derived search terms, we used the web-based Virtual Observatory for the Study of Online Networks (VOSON) to conduct hyperlink network analysis (HNA) of suicide-related websites. RESULTS: Pro-suicide sites are rare and marginal, while sites dedicated to information about suicide as well as sites dedicated to prevention policy and advocacy are readily accessible. CONCLUSIONS: The networking structure of suicide related Internet content has not been described previously. Our analysis shows that HNA is a useful method for gaining an indepth understanding of network traffic in relation to suicide-content websites. This information will be useful for strengthening the web presence of support and suicide prevention sites, and for monitoring changes over time. PMID- 21616764 TI - The economic and potential years of life lost from suicide in Taiwan, 1997-2007. AB - BACKGROUND: Taiwan has experienced a marked increase in the suicide rate in the last decade. However, the socioeconomic burden and impact to the community has not been adequately assessed. AIMS: This study aimed to estimate the social and economic burden of premature mortality from suicide in Taiwan in 1997-2007. METHODS: The suicide rate, potential years of life lost (PYLL), and present value of lifetime earnings (PVLE) by sex and age groups in 1997-2007 were calculated. The contribution of each suicide method to PYLL for each age group was also assessed. RESULTS: Using the PYLL calculations, suicide had become the third leading cause of death in Taiwan in 2007, compared to its ninth position in terms of absolute numbers. Furthermore, the PYLL was associated with an estimated NTD (New Taiwan Dollars) 32.5 billion of lost earnings in 2007. The increase in PYLL and PVLE from suicide was highest in middle-aged men (aged 25-59 years). Charcoal burning suicide accounted for most of the increase in PYLL in the middle-aged group in the past decade. CONCLUSIONS: The loss of life in middle-aged males contributes disproportionately to the social and economic burden of suicide in Taiwan. Suicide intervention effort should target this high-risk population. PMID- 21616765 TI - Integrating medical examiner and police report data: can this improve our knowledge of the social circumstances surrounding suicide? AB - BACKGROUND: Recently, suicide in the United States has begun to be viewed as a preventable public health issue. This has led to the creation of a National Violent Death Reporting System that collects and integrates data on the social circumstances surrounding suicides. AIMS: The study examines data on social circumstances surrounding suicides as collected by the medical examiner report (ME) and police report (PR) and subsequently integrated into the state of Maryland's violent death reporting system. METHODS: Reported data on social circumstances surrounding suicides occurring in the years 2003-2006 in Maryland (n = 1,476) were analyzed by examining their prevalence in the ME and PR, strength of association, and integration. RESULTS: With the exception of three circumstances, there was variation among reported circumstances in the ME and PR. Furthermore, there was only a moderately strong relationship between the ME and PR for most circumstances, while a significant increase occurred in the prevalence of these circumstances when ME and PR were integrated. CONCLUSIONS: The integration of ME and PR has the potential to increase our knowledge of the circumstances surrounding suicide and to better inform prevention efforts. However, before this potential can be reached, there are still issues that must be considered. PMID- 21616766 TI - Panic disorder and suicidal behavior: a follow-up study of patients treated with cognitive therapy and SSRIs in Hungary. AB - BACKGROUND: Previous research has suggested that patients with panic disorder but no comorbid disorder are not at greater risk for suicidal behavior. AIMS: The present study followed up patients with panic disorder in order to assess the frequency of their suicidal behavior. METHODS: A sample of 281 outpatients with panic disorder, but without a comorbid psychiatric disorder, was followed up for an average of 5 years. The patients were given 6-8 weeks of cognitive therapy, and 65% were prescribed SSRIs. RESULTS: At the time of first admission, 5 patients (1.7%) reported a previous (lifetime) suicide attempt, and 53 patients (18.2%) reported previous (lifetime) suicidal ideation (both thoughts and plans), not greatly different from the Hungarian population in general. During the follow up period, no patient committed suicide, 2 patients attempted suicide (0.7%), and 4 patients (1.4%) reported suicidal ideation. CONCLUSIONS: This study indicates that people with panic disorders without comorbid disorders have no higher suicidal risk than the general population in Hungary. After treatment with cognitive therapy and SSRIs, 38.5% were symptom-free, and only 7.8% required continued close therapeutic contact after the follow-up period. PMID- 21616768 TI - Numerical alterations of ageing B lymphocyte subsets. AB - The global population is ageing. Elderly people suffer from more severe infections than younger persons. The major reason for the increased susceptibility to infections in the elderly is the deregulated functions of the immune system. Immunosenescence affects both innate and adaptive immune reactions. Among these, quantitative alterations of B lymphocyte subsets determine outcome of infections and vaccination. The overall number of B cells seems to be stable or the decrease is moderate. Reduced input of naive B lymphocytes is compensated by anergic, exhausted memory cells. Concerning B lymphocyte subsets, experimental data obtained in the mouse model and in vivo studies conducted in old-age humans are frequently controversial. Further analysis of human B lymphocyte subpopulations is required that could be regarded as an important biomarker of human life span. PMID- 21616769 TI - The importance of body composition and dry weight assessments in patients with chronic kidney disease. AB - Chronic volume overload is the major cause of hypertension and other cardiovascular morbidity in dialysis patients. One of the most important goals of physicians who take care of patients with chronic renal failure is to obtain near euvolemia or "dry body weight" in order to maintain or normalize blood pressure and prevent further cardiovascular events. In clinical practice, exact estimation of dry weight in hemodialysis patients remains a major challenge. Alterations in body composition, particularly malnutrition, are common in patients receiving long-term hemodialysis and contribute to a high mortality rate. In contrast, obesity - a known risk factor for cardiovascular morbidity and mortality - is prevalent amongst kidney allograft recipients in - long term after renal transplantation. Several technological tools and biochemical markers for estimation of plasma volume and body composition are available for clinical use. Our aim was to highlight the importance of control of body fluid volume and body composition in patients with chronic kidney disease and to describe the different methods available for such measurements. PMID- 21616770 TI - Carotid arterial stiffness in patients with chronic obstructive pulmonary disease. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is an important cause of morbidity and mortality.Patients with COPD have a cardiovascular risk three times greater than the normal population and cardiac disease accounts for about 50% of deaths in these patients. Arterial stiffness is a marker of increased cardiovascular risk and a predictor for cardiovascular morbidity and mortality. There is evidence of increased aortic stiffness in COPD. The aim of our study is to determine parameters of carotid arterial stiffness in patients with stable COPD. PATIENTS AND METHODS: We enrolled 38 patients with COPD and 36 matched controls. All subjects were evaluated by clinical history and spirometry; parameters of carotid stiffness (beta stiffness index - 13 stiffness index, arterial compliance - AC, pulse wave velocity - PWV, augmentation index - AIx) were determined using an ultrasound device (ALOKA ProSound a 10). RESULTS: Patients with COPD have increased markers of arterial stiffness compared to controls (B stiffness index,p=0.03; AC, p=0.002; carotid PWV, p<0.001; Alx, p<0.001). There were no differences between the two groups regarding the classical cardiovascular risk factors with the exception of smoking history. Patients with severe pulmonary disease had more increased 13 stiffness index and PWV than those with mild and moderate disease(p=0.03, respectively p=0.004). CONCLUSIONS: Patients with COPD had increased carotid arterial stiffness. In severe forms of COPD, arterial stiffness seems to be more important than in mild and moderate pulmonary disease. PMID- 21616771 TI - Exercise training biomarkers: influence of short-term diet modification on the blood lactate to rating of perceived exertion (La:RPE) ratio. AB - This study examined the effect of dietary consumption of carbohydrates (CHO) on the blood lactate to rating of perceived exertion (La:RPE) ratio during an intense micro-cycle of exercise training. This ratio is a proposed biomarker of exercise training stress and potential indicator for under- or overtraining in athletes. Sixteen male athletes were randomly assigned into two groups; high CHO (H-CHO; 60% of daily caloric intake) and low CHO (L-CHO; 30% of daily caloric intake). Diets were controlled the day before and for the three days of the micro cycle. The micro-cycle consisted of three successive days of 60 minutes of intense cycling (~70% of VO2peak). Blood samples were obtained immediately before and after exercise (post) on each day of exercise training (D1, D2, D3) and were analyzed for blood lactate. Rating of perceived exertion (RPE) scores were taken at the end of each exercise session and combined with the post exercise lactate value to form the La:RPE ratio. An analysis of variance (ANOVA) showed a significant difference between the La:RPE ratio for the H-CHO and L-CHO groups at D3 even though the exercise intensity was not significantly different between the groups. Specifically, the ratio was significantly (p < 0.02) lower on D3 in the L CHO group (~31% lower) than in the H-CHO group. From these findings it is recommended that diet needs to be monitored when using the La:RPE ratio as an exercise training biomarker to determine whether an athlete is truly under training or overtraining. Athletes or coaches that use the La:RPE ratio as a training biomarker, but do not monitor dietary CHO intake need to interpreted their findings carefully. PMID- 21616772 TI - Left atrial and left atrial appendage function in paroxysmal atrial fibrillation. AB - PURPOSE: In patients with paroxysmal atrial fibrillation (PAF) little information is available about left atrial (LA)function, and there is less information about LA appendage (LAA) function, and about their relations. METHODS AND RESULTS: 46 patients were selected for catheter ablation (CA) because of nonvalvular PAF.Transthoracic, tissue Doppler and transoesophageal echocardiography was performed before CA. LA volumes and volume index (LAVI) were calculated. LA function was assessed by LA filling fraction (LAFF), LA emptying fraction (LAEF), systolic fraction of pulmonary venous flow (PVSF) and late diastolic velocities of mitral annulus(Aa,, A5at) LAA function was assessed by peak LAA emptying flow velocity (PLAAEFV). Diastolic dysfunction(DD) was also assessed. Dilated LAVI in 32, LA dysfunction in 20, DD with elevated LV filling pressure in 19 patients was found. Aa,at and Aa,p correlated with LAFF (r:0.53; p<0.001 and r:0.43; p<0.05), LAEF (r:0.51;p<0.001 and r:0.63; p<0.001), PVSF (r:0.49; p<0.001 and r:0.46; p<0.005) and PLAAEFV (r:0.58; p<0.001 and r:0.45; p<0.01). CONCLUSIONS: In PAF patients Aa velocity is useful to assess LA function and correlates positively with other TTE derived LA functional parameters and LAA function by TEE derived PLAAEFV. PMID- 21616773 TI - The validity of incremental exercise testing in discriminating of physiological profiles in elite runners. AB - The goal of this study was to determine whether traditional ergoespirometric incremental exercise testing carried out to the point of exhaustion could be useful in distinguishing the physiological profiles of elite runners that compete in races that lasted about 8 minutes versus those that lasted about 2 hours. Ten male marathon runners (performance time: 2:12:04, coefficient of variation (CV) = 2.33%) and 8 male 3000 m steeplechase runners (performance time: 8:37.83, CV = 2.12%) performed an incremental test on the treadmill (starting speed 10 km.h-1; increments, 2 km.h-1; increment duration, 3 min to exhaustion). Heart rate (HR), VO2, and lactate concentrations were measured at the end of each exercise level. At maximal effort, there were no differences between the groups regarding VO2max and maximal HR; however, the workload time, vVO2max and peak treadmill velocity were significantly higher in the 3000 m steeplechase group (p<0.05). At submaximal effort, there were no significant differences between groups for VO2 (ml.kg-1.min-1), HR, or lactate. Our results show that this type of testing was not sufficient for discriminating the physiological profiles of elite runners who competed in middle-distance versus long-distance events (e.g. in the marathon and the 3000 m steeplechase). PMID- 21616774 TI - Involuntary human hand movements due to FM radio waves in a moving van. AB - Finland TRACT Involuntary movements of hands in a moving van on a public road were studied to clarify the possible role of frequency modulated radio waves on driving. The signals were measured in a direct 2 km test segment of an international road during repeated drives to both directions. Test subjects (n=4) had an ability to sense radio frequency field intensity variations of the environment. They were sitting in a minivan with arm movement detectors in their hands. A potentiometer was used to register the hand movements to a computer which simultaneously collected data on the amplitude of the RF signal of the local FM tower 30 km distance at a frequency of about 100 MHz. Involuntary hand movements of the test subjects correlated with electromagnetic field, i.e. FM radio wave intensity measured. They reacted also on the place of a geomagnetic anomaly crossing the road, which was found on the basis of these recordings and confirmed by the public geological maps of the area.In conclusion, RF irradiation seems to affect the human hand reflexes of sensitive persons in a moving van along a normal public road which may have significance in traffic safety. PMID- 21616775 TI - Involvement of D1 receptors in depression-like behavior of ovariectomized rats. AB - The aim of the present study was to explore the mood effects of D1 receptor agonist, SKF-38393 and D1 receptor antagonist, SCH-23390 alone or in combination with a low dose of 17beta-estradiol (17beta-E2) in the adult ovariectomized female rats (OVX). OVX rats of Wistar strain were used in all experiments. Two weeks after surgery rats were chronically treated with vehicle, a low dose of 17beta-E2 (5.0 MUg/rat), SKF-38393 (0.1 mg/kg), SCH-23390 (0.1 mg/kg), SKF-38393 plus 17beta-E2 or SCH-23390 plus 17beta-E2 for 14 days before the forced swimming test. We found that SCH-23390 significantly decreased immobility time in the OVX females. A combination of SCH-23390 with a low dose of 17beta-E2 induced more profound decrease of immobility time in the OVX rats compared to the rats treated with SCH-23390 alone. On the contrary, SKF-38393 failed to modify depression-like behavior in the OVX rats. In addition, SKF-38393 significantly blocked the antidepressant-like effect of 17beta-E2 in OVX rats. Thus, the D1 receptor antagonist SCH-23390 alone or in combination with a low dose of 17beta-E2 exerted antidepressant-like effect in OVX female rats, while the D1 receptor agonist SKF 38393 produced depressant-like profile on OVX rats. PMID- 21616776 TI - Monosodium glutamate versus diet induced obesity in pregnant rats and their offspring. AB - BACKGROUND: We aim at determining the role of monosodium glutamate (MSG) compared with high caloric chow(HCC) in development of obesity in pregnant rats and their offspring. METHODS: Ninety pregnant rats were divided into 3 groups, control, MSG and HCC fed. We determined energy intake, body weight (BW), abdominal fat, fat to body weight ratio, serum glucose, insulin, leptin, lipid profile, ob and leptin receptor-b gene expressions in pregnant rats and ob and leptin receptor-b gene expressions, serum insulin,glucose, leptin, triacylglycerides (TAG), total lipids (TL) and BW in offspring. RESULTS: Although daily energy intake and BW of MSG treated rats were lower than those of HCC fed rats, their abdominal fat and fat body weight ratio were higher. MSG or HCC increased Ob gene expression, leptin, insulin,LDL, cholesterol, total lipids (TL), glucose and decreased leptin receptor-b gene expression. In offspring of MSG treated rats, BW, serum glucose, insulin, leptin, TAG, TL and Ob gene expression increased and leptin receptor-b gene expression decreased whereas in offspring of HCC fed rats, serum insulin, leptin, Ob and leptin receptor-b gene expression increased but serum glucose, TAG, TL or BW did not change. CONCLUSION: We conclude that in pregnant rats, MSG, in spite of mild hypophagia, caused severe increase in fat body weight ratio, via leptin resistance, whereas, HCC increased BW and fat body weight ratio, due to hyperphagia with consequent leptin resistance. Moreover, maternal obesity in pregnancy, caused by MSG, has greater impact on offspring metabolism and BW than that induced by HCC. PMID- 21616777 TI - Single, high-dose 17beta-estradiol therapy has anti-apoptotic effect and induces cerebral plasticity following transient forebrain ischemia in gerbils (Short communication). AB - Although much is known about the protective effect of acute estrogen therapy in cerebral ischemia, relatively little is known about the importance of apoptosis and cerebral plasticity in this mechanism. In this work 10 min global cerebral ischemia was produced by transient bilateral carotid occlusion in 4-month-old ovariectomized female gerbils. In every of our experimental group (sham for ischemia group, ischemia group and ischemia + a high, single dose 17beta estradiol pre-treatment group) apoptotic (bcl-Xl, bax) and cerebral plasticity (GAP-43, synapsin-I, nestin) hippocampal genes' expression was measured four days after surgery. Expression of the anti-apoptotic bcl-Xl (p<0.01) and the cerebral plasticity marker synapsin-I and nestin (p<0.01) increased with acute estrogen pretreatment in ischemic animals. No change, however, in bax or GAP-43 expression was detected in estrogen treated animals compared to ischemic gerbils. These results suggest that acute estrogen therapy has anti-apoptotic effect and increases cerebral plasticity, which play an important role in cytoprotection or cerebroprotection. PMID- 21616778 TI - Non-invasive oxidative stress markers for liver fibrosis development in the evolution of toxic hepatitis. AB - Oxidative stress is related to the liver fibrosis, anticipating the hepatic stellate cells' (HSC) activation. Our aim was to correlate oxidative stress markers with the histological liver alterations in order to identify predictive, noninvasive parameters of fibrosis progression in the evolution of toxic hepatitis.CCl4 in sunflower oil was administered to rats intragastrically, twice a week. After 2, 3, 4 and 8 weeks of treatment, plasma levels of malondialdehyde (MDA), protein carbonyls (PC), hydrogen donor capacity (HD), sulfhydryl groups (SH), and glutathione (GSH) were measured and histological examination of the liver slides was performed. Dynamics of histological disorders was assessed by The Knodell score. Significant elevation of inflammation grade was obtained after the second week of the experiment only (p=0.001), while fibrosis started to become significant (p=0.001) after 1 month of CCl4 administration. Between plasma MDA and liver fibrosis development a good correlation was obtained (r=0.877, p=0.05). Correlation between PC dynamics and liver alterations was marginally significant for inflammation grade (r=0.756, p=0.138). HD evolution revealed a marginally inverse correlation with inflammation grade (r=-0.794, p=0.108). No correlations could be established for other parameters with either inflammation grade or fibrosis stage.Our study shows that MDA elevation offers the best prediction potential for fibrosis, while marginal prediction fiability could be attributed to high levels of plasma PC and low levels of HD. PMID- 21616779 TI - Effects of fluoxetine and LY 367265 on tolerance to the analgesic effect of morphine in rats. AB - Morphine is widely used to treat chronic pain, however its utility is hindered by the development of tolerance to its analgesic effects. The aim of this study was to investigate effects of fluoxetine, a specific serotonin (5-HT) reuptake inhibitor, and LY 367265, an inhibitor of the 5-HT transporter and 5-HT2A receptor antagonist, on tolerance induced to the analgesic effect of morphine in rats. The study was carried out on male Wistar Albino rats (weighing 170-190 g). To constitute morphine tolerance, animals received morphine (50 mg/kg; s.c.) once daily for 3 days. After last dose of morphine, injected on day 4, morphine tolerance was evaluated. The analgesic effects of fluoxetine (10 mg/ kg; i.p.), LY 367265 (3 mg/kg; i.p.) and morphine were considered at 30-min intervals by tail-flick and hot-plate tests. The results showed that fluoxetine and LY 367265 significantly attenuated the development and expression of morphine tolerance. The maximal antinociceptive effects were obtained 30 min after administration of fluoxetine and 60 min after administration of LY 367265. In conclusion, we observed that co-injection of morphine with fluoxetine and LY 367265 increased the analgesic effects of morphine and delayed development of tolerance to morphine analgesia. PMID- 21616780 TI - Effect of vitamin E therapy on serum uric acid in DOCA-salt-treated rats. AB - Uric acid is considered as an antioxidant in the blood. Despite its proposed protective properties, elevated plasma uric acid has been associated with hypertension in a variety of disorders. The purpose of this study was to investigate the relationship between the increase of arterial blood pressure and the changes in serum uric acid, measured during the gradual development of experimental hypertension in deoxycorticosterone (DOCA)-salt-treated rats. Blood pressure was monitored by tail-cuff method, urinary and plasma uric acid was measured by autoanalyzer during the induction of hypertension in 1-, 2-, 3- and 4 week DOCA-salt-treated Sprague-Dawley rats. Vitamin E (200 mg/kg/day/gavage) was co-administered with DOCA-salt for 4 weeks. From the first week of DOCA-salt treatment, rats exhibited marked increases in blood pressure. DOCA-salt treatment also resulted in a significant increase in serum uric acid and a significant decrease in urinary uric acid at the end of the first week. These changes in serum and urinary uric acid remained until the 4th week of DOCA-salt treatment but blood pressure continued to increase throughout the study. Vitamin E treatment increased urinary excretion of uric acid and decreased blood pressure and serum uric acid in DOCA-salt-treated rats. These data suggest that enhanced serum uric acid may be a contributing factor to the onset of hypertension in DOCA salt-treated rats. A uricosuric effect is suggested for vitamin E in the treatment of hypertension. PMID- 21616781 TI - Glabridin as a major active isoflavan from Glycyrrhiza glabra (licorice) reverses learning and memory deficits in diabetic rats. AB - Cognitive impairment occurs in diabetes mellitus. Glabridin as a major active flavonoids in Glycyrrhiza glabra (licorice) improves learning and memory in mice. In the present study, we investigated the effect of chronic treatment with glabridin (5, 25 and 50 mg/kg, p.o.) on cognitive function in control and streptozotocin (STZ)-induced diabetic rats.Animals were divided into untreated control, glabridin-treated control (5, 25 and 50 mg/kg), untreated diabetic and glabridin treated diabetic (5, 25 and 50 mg/kg) groups. Treatments were begun at the onset of hyperglycemia. Passive avoidance learning (PAL) and memory was assessed 30 days later. Diabetes caused cognition deficits in the PAL and memory paradigm. While oral glabridin administration (25 and 50 mg/kg) improved learning and memory in non-diabetic rats, it reversed learning and memory deficits of diabetic rats. Low dose glabridin (5 mg/kg) did not alter cognitive function in non-diabetic and diabetic groups. Glabridin treatment partially improved the reduced body weight and hyperglycemia of diabetic rats although the differences were not significant. The combination of antioxidant, neuroprotective and anticholinesterase properties of glabridin may all be responsible for the observed effects. These results show that glabridin prevented the deleterious effects of diabetes on learning and memory in rats. Further studies are warranted for clinical use of glabridin in the management of demented diabetic patients. PMID- 21616782 TI - Blood oxidative stress generation after intraperitoneal administration of functionalized single-walled carbon nanotubes in rats. AB - Single-walled carbon nanotubes (SWCNTs) have been proposed for various medical applications. However, their safety for human administration has not been yet fully demonstrated. In vitro studies have pointed oxidative stress as a mechanism involved in their cytotoxic effects. In the present study we have evaluated the capacity of DNA functionalized SWCNTs to induce oxidative stress in blood after intraperitoneal (ip) administration in rats. The presence of SWCNTs in blood was confirmed by Raman spectroscopy 30 minutes after their ip administration. Oxidative stress parameters (malondialdehyde - MDA, protein carbonyls - PC, antioxidant capacity measured as hydrogen donating capacity - HD, sulfhydryl groups - SH, glutathione - GSH and nitrites - NO) were assessed in blood at 3, 6, 24, respectively, and 48 hours after ip injection. MDA, PC and NO exhibited a significant increase at 3-6 hours interval from exposure, followed by a recovery trend. The levels of HD reached a bottom level at 6 hours after administration, while SH strongly decreased at 3 hours interval and increased slightly up to 48 hours without attending the initial values. GSH level recorded an increasing tendency at the 3rd hour, an incomplete recovery process at 24 hours followed by a secondary significant increase following a 48-hour interval. Significant inverse correlations were obtained between the PC and SH levels and between the NO and HD values. In conclusion, the ip administration of DNA functionalized SWCNT in rats results in oxidative stress generation in plasma, with a transient pattern of evolution. PMID- 21616784 TI - Impact of health portal enrollment with email reminders on adherence to clinic appointments: a pilot study. AB - BACKGROUND: Internet portal technologies that provide access to portions of electronic health records have the potential to revolutionize patients' involvement in their care. However, relatively few descriptions of the demographic characteristics of portal enrollees or of the effects of portal technology on quality outcomes exist. This study examined data from patients who attended one of seven Duke Medicine clinics and who were offered the option of enrolling in and using the Duke Medicine HealthView portal (HVP). The HVP allows patients to manage details of their appointment scheduling and provides automated email appointment reminders in addition to the telephone and mail reminders that all patients receive. OBJECTIVE: Our objective was to test whether portal enrollment with an email reminder functionality is significantly related to decreases in rates of appointment "no-shows," which are known to impair clinic operational efficiency. METHODS: Appointment activity during a 1-year period was examined for all patients attending one of seven Duke Medicine clinics. Patients were categorized as portal enrollees or as nonusers either by their status at time of appointment or at the end of the 1-year period. Demographic characteristics and no-show rates among these groups were compared. A binomial logistic regression model was constructed to measure the adjusted impact of HVP enrollment on no-show rates, given confounding factors. To demonstrate the effect of HVP use over time, monthly no-show rates were calculated for patient appointment keeping and contrasted between preportal and postportal deployment periods. RESULTS: Across seven clinics, 58,942 patients, 15.7% (9239/58,942) of whom were portal enrollees, scheduled 198,199 appointments with an overall no show rate of 9.9% (19,668/198,199). We found that HVP enrollees were significantly more likely to be female, white, and privately insured compared with nonusers. Bivariate no-show rate differences between portal enrollment groups varied widely according to patient- and appointment-level attributes. Large reductions in no-show rates were seen among historically disadvantaged groups: Medicaid holders (OR = 2.04 for nonuser/enrollee, 5.6% difference, P < .001), uninsured patients (OR = 2.60, 12.8% difference, P < .001), and black patients (OR = 2.13, 8.0% difference, P < .001). After fitting a binomial logistic regression model for the outcome of appointment arrival, the adjusted odds of arrival increased 39.0% for portal enrollees relative to nonusers (OR = 1.39, 95% CI 1.22 - 1.57, P < .001). Analysis of monthly no-show rates over 2 years demonstrated that patients who registered for portal access and received three reminders of upcoming appointments (email, phone, and mail) had a 2.0% no show rate reduction (P < .001), whereas patients who did not enroll and only received traditional phone and mail reminders saw no such reduction (P < .09). CONCLUSIONS: Monthly no-show rates across all seven Duke Medicine clinics were significantly reduced among patients who registered for portal use, suggesting that in combination with an email reminder feature, this technology may have an important and beneficial effect on clinic operations. PMID- 21616786 TI - Microsatellite primers in the Peltigera dolichorhiza complex (lichenized ascomycete, Peltigerales). AB - PREMISE OF THE STUDY: Microsatellite primers were developed for the lichen forming fungus Peltigera dolichorhiza to investigate partitioning of genetic variation in a widespread, morphologically and chemically variable taxon likely to represent a complex of cryptic lineages, including P. neopolydactyla. * METHODS AND RESULTS: Using next generation shotgun sequence reads, 331 primer pairs were designed to amplify microsatellite sequences from an African accession of P. dolichorhiza. Eleven primer pairs representing the longest repeat units identified were tested on 15 P. dolichorhiza accessions from Africa (incl. Reunion), South America, Papua New Guinea, and on two accessions of P. neopolydactyla from North America. The primers amplified di-, tri, tetra-, and pentanucelotide repeats with 3-8 alleles per locus. All individuals represent distinct multiloci genotypes. * CONCLUSIONS: These results indicate the utility of the new microsatellite primers for testing genetic differentiation within the widespread complex of P. dolichorhiza. PMID- 21616787 TI - Polymorphic chloroplast microsatellite markers in the octoploid Lepidium meyenii (Brassicaceae) and cross-species amplification in Lepidium. AB - PREMISE OF THE STUDY: As a crop and medicinal plant, the octoploid Andean endemic Lepidium meyenii suffers from taxonomic uncertainty. Few molecular markers are available to genotype individuals or track gene flow in wild and cultivated material. * METHODS AND RESULTS: Using available sequence data, eight cpSSR primer pairs were developed for L. meyenii. Levels of polymorphism checked in 56 individual L. meyenii, including cultivated and wild material, revealed that the number of alleles per locus ranged from three to five, and intrapopulation allele frequencies ranged from 0.071 to 1.0. Polymerase-chain reaction screens using our cpSSR primers in 27 other Lepidium species and three Coronopus species suggested a high degree of interspecific amplification. * CONCLUSIONS: These polymorphic cpSSR markers should prove useful in characterizing genetic variation among cultivated and wild L. meyenii. Additionally, interspecific amplifications suggest that these markers will be useful for the study of related taxa. PMID- 21616788 TI - Isolation and characterization of polymorphic microsatellite loci in Camellia nitidissima Chi (Theaceae). AB - PREMISE OF THE STUDY: Microsatellite markers were developed in Camellia nitidissima for further population genetic studies. * METHODS AND RESULTS: Eight microsatellite markers were newly developed from C. nitidissima and 7 were transferred from other Camellia species. Two to 13 alleles per locus were identified for these microsatellites. Observed and expected heterozygosities ranged from 0.040 to 0.909, and 0.184 to 0.916, respectively. Four loci showed a significant deviation from Hardy-Weinberg equilibrium and five locus pairs displayed linkage disequilibrium. * CONCLUSIONS: These microsatellite markers will be useful to assess the genetic variation and genetic structure of C. nitidissima. PMID- 21616789 TI - Isolation and characterization of microsatellite markers in an endangered species Dracaena cambodiana (Liliaceae). AB - * PREMISE OF THE STUDY: The development of microsatellite primers in the endangered species Dracaena cambodiana will be the foundation for genetic and conservation studies of D. cambodiana and several Dracaena species. * METHODS AND RESULTS: A total of 26 microsatellite markers were developed in Chinese populations of D. cambodiana, using the Fast Isolation by AFLP of Sequences Containing Repeats (FIASCO) protocol. Among them, sixteen primer pairs generated polymorphic loci (fourteen of them successfully amplified in other four Dracaena species) and ten primer pairs produced monomorphic loci. * CONCLUSIONS: These microsatellite markers could be used in the further investigation of population genetics of D. cambodiana and other Dracaena species. PMID- 21616790 TI - Microsatellite markers for the cushion rock jasmine, Androsace tapete (Primulaceae), a species endemic to the Qinghai-Tibetan Plateau. AB - PREMISE OF THE STUDY: Microsatellite markers were developed in an alpine plant endemic to the Qinghai-Tibetan Plateau, Androsace tapete, to investigate its spatial genetic structure, gene flow, and mating patterns. * METHODS AND RESULTS: Using the combined biotin capture method, 16 microsatellite primer sets were isolated and characterized. Fifteen of these markers showed polymorphism, and the number of alleles per locus ranged from three to 13 across 56 individuals from six Tibetan populations. * CONCLUSIONS: These markers provide a useful tool to investigate the spatial genetic structure, gene flow, and mating system of A. tapete. PMID- 21616791 TI - Isolation and characterization of microsatellite loci in Santalum lanceolatum and Santalum leptocladum (Santalaceae). AB - PREMISE OF THE STUDY: Microsatellite primers were developed for the first time in the native Australian sandalwood species Santalum lanceolatum. * METHODS AND RESULTS: Using an enrichment cloning protocol, five novel polymorphic codominant loci were developed and characterized in S. lanceolatum and S. leptocladum. In addition to these, three existing microsatellite loci from other sandalwood species were successfully amplified and characterized for S. lanceolatum and S. leptocladum. Among the eight loci, allelic diversity ranged from 4 to 29. * CONCLUSIONS: Primers will be useful for studies of clonality, genetic diversity and spatial genetic structure in wild populations. When coupled with other molecular techniques will help investigate the relationship between S. lanceolatum and S. leptocladum, species of commercial and conservation interest. PMID- 21616792 TI - Characterization and multiplexing of EST-SSR primers in Cynodon (Poaceae) species1. AB - PREMISE OF THE STUDY: Cynodon species are multiple-use grasses that display varying levels of adaptation to biotic and abiotic stress. Previously identified EST-SSR primers were characterized and multiplexed to assess the level of genetic diversity present within a collection of almost 1200 Cynodon accessions from across Australia. * METHODS AND RESULTS: Two multiplex reactions were developed comprising a total of 16 EST-SSR markers. All SSR markers amplified across different Cynodon species and different levels of ploidy. The number of alleles ranged from one to eight per locus and the total number of alleles for the germplasm collection was 79. * CONCLUSIONS: The 16 markers show sufficient variation for the characterization of Cynodon core collections and analysis of population genetic diversity in Cynodon grasses. PMID- 21616793 TI - Worldwide correlations of mechanical properties and green wood density. AB - PREMISE OF THE STUDY: The density of wood is highly correlated with the ability of stems and roots to resist bending or twisting, which is important for evaluating the mechanical behavior of trees. It also provides a measure of carbon storage, which is an important variable in modeling ecosystem processes and tree construction costs. However, most measurements of the density and mechanical properties of wood have little direct bearing on understanding the biomechanics of living plants because they are based on kiln- or air-dried samples. * METHODS: Here, we present and analyze the relationships between four important mechanical properties (Young's modulus, the modulus of rupture, and the maximum strength in shearing and in compression) and the density of green wood (i.e., wood at 50% moisture content) from a worldwide, taxonomically broad spectrum of 161 species. * KEY RESULTS: These data indicate that each of the mechanical properties disproportionately increases across species with increasing green wood density, i.e., stems composed of denser green wood are disproportionately stiffer and stronger than stems with equivalent cross-sections composed of less dense green wood. * CONCLUSIONS: Although denser wood may have a higher carbon construction cost, the mechanical benefits of denser woods likely outweigh the extra cost. PMID- 21616794 TI - The seed dispersal catapult of Cardamine parviflora (Brassicaceae) is efficient but unreliable. AB - PREMISE OF THE STUDY: Seed dispersal performance is an essential component of plant fitness. Despite their significance in shaping performance, the mechanical processes that drive dispersal are poorly understood. We have quantified seed dispersal mechanics in Cardamine parviflora (Brassicaceae), a ballistic disperser that launches seeds with specialized catapult-like structures. To determine which aspects of catapult function dictate interspecific dispersal differences, we compared this disperser with other ballistic dispersers. Comparison with brassicas that lack ballistic dispersal may also provide insight into the evolution of this mechanism. * METHODS: Catapult performance was quantified using high-speed video analysis of dehiscence, ballistic modeling of seed trajectories, and measuring the mechanical energy storage capacity of the spring-like siliqua valve tissue that launched the seeds. * KEY RESULTS: The siliquae valves coiled rapidly outward, launching the seeds in 4.7 +/- 1.3 ms (mean +/- SD, N = 11). Coiling was likely driven by the bilayered valve structure. The catapult was 21.3 +/- 10.3% efficient (mean +/- SD, N = 11) at transferring stored elastic energy to the seeds as kinetic energy. The majority of seeds (71.4%) were not launched effectively. * CONCLUSIONS: The efficiency of the C. parviflora catapult was high in comparison to that of a ballistic diplochore, a dispersal mode associated with poor ballistic performance, although the unreliability of the launch mechanism limited dispersal distance. Effective launching requires temporary seed-valve adhesion. The adhesion mechanism may be the source of the unreliability. Valve curvature is likely driven by the bilayered valve structure, a feature absent in nonballistic brassicas. PMID- 21616795 TI - Induction of programmed cell death in Arabidopsis and rice by single-wall carbon nanotubes. AB - PREMISE OF THE STUDY: Single-walled carbon nanotubes (SWCNTs) have many unique structural and mechanical properties. Their potential applications, especially in biomedical engineering and medical chemistry, have been increasing in recent years, but the toxicological impact of nanoparticles has rarely been studied in plants. * METHODS: We exposed Arabidopsis and rice leaf protoplasts to SWCNTs and examined cell viability, DNA damage, reactive oxygen species generation, and related gene expression. We also tested the effects of nanoparticles on Arabidopsis leaves after injecting a SWCNT solution. EM-TUNEL (electron microscopic terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling) and a cerium chloride staining method were used. * KEY RESULTS: SWCNTs caused adverse cellular responses including cell aggregation, chromatin condensation along with a TUNEL-positive reaction, plasma membrane deposition, and H(2)O(2) accumulation. The effect of SWCNTs on the survival of cells was dose dependent, with 25 MUg/mL inducing 25% cell death in 6 h. In contrast, activated carbon, which is not a nano-sized carbon particle, did not induce cell death even 24 h after treatments. The data indicated that the nano-size of the particle is a critical factor for toxicity. Moreover, endocytosis-like structures with cerium chloride deposits formed after SWCNT treatment, suggesting a possible pathway for nanoparticles to traverse the cell membrane. * CONCLUSIONS: Consequently, SWCNTs have an adverse effect on protoplasts and leaves through oxidative stress, leading to a certain amount of programmed cell death. Although nanomaterials have great advantages in many respects, the benefits and side effects still need to be assessed carefully. PMID- 21616796 TI - Growth and fitness components of wild x cultivated Sorghum bicolor (Poaceae) hybrids in Nebraska. AB - PREMISE OF THE STUDY: Gene flow from crops to wild relatives has received considerable attention since the advent of genetically modified crops. Numerous researchers have found wild-crop hybrids to be nearly as fit as their wild parents, which suggests that crop genes may persist in wild populations. Components of the ecological fitness of cultivated sorghum, its wild relative, shattercane, and their hybrids have not been studied. * METHODS: To assess the potential for gene introgression into shattercane, we crossed cultivated sorghum to a single inbred shattercane line to produce F(1) hybrids and measured growth and several components of ecological fitness in relation to both parents in Nebraska, USA. * KEY RESULTS: Germination of F(1) seeds was similar to that of its shattercane parent except at high temperatures, where it was as sensitive as the sorghum parent. The F(1) grew taller and produced more biomass than either parent, but the F(1) leaf area index was intermediate. Fecundity of the F(1) plant was similar to that of shattercane and much greater than that of cultivated sorghum. * CONCLUSIONS: Considering all data, the ecological fitness of shattercane * cultivated sorghum F(1) hybrids may be equivalent to the wild shattercane parent, which suggests that crop genes that are either neutral or beneficial to shattercane would persist in populations within agroecosystems. PMID- 21616797 TI - Change in floral orientation in Anisodus luridus (Solanaceae) protects pollen grains and facilitates development of fertilized ovules. AB - PREMISE OF THE STUDY: Some floral traits could be selected by pollinators and nonpollinator agents, and studying the floral traits shaped by physical agents could reveal adaptive mechanisms to the environment. We explored the adaptive significance of the change in floral orientation, from pendulous flowers to erect fruits that have a persistent calyx, in Anisodus luridus, a perennial native to the Qinghai-Tibet Plateau (QTP). * METHODS: We examined the effect of flower orientation on pollinator visitation rates, pollen deposition efficiency, pollen quantity, and pollen viability to estimate whether pendulous flowers have improved male fitness. We then measured seed production and seed germination rate to assess whether erect fruits have enhanced female fitness. * KEY RESULTS: Pendulous flowers did not have any preferred pollinators or increased pollen deposition. In artificially erected flowers, the number of pollen grains greatly decreased after rainwash. Pollen germination experiments indicated that pollen damage by water and exposure to solar radiation is serious. In the persistent calyx that holds water within it, the temperature inside the calyx changed slower than in the calyx that had the water removed. After supplemental pollination, the seed number, seed set, and seed mass of fruits that had water removed from the calyx were reduced greatly in both years. * CONCLUSIONS: We conclude that the change in floral orientation could enhance male and female fitness of A. luridus and is effectively adaptive to the alpine environments, indicating a strong selection by the combined pressure from various abiotic nonpollinator agents in shaping the floral traits of this alpine plant. PMID- 21616798 TI - The impact of altered precipitation variability on annual weed species. AB - PREMISE OF THE STUDY: Climate change models predict increasing variability in precipitation across the globe, with an increase in the incidence of large precipitation events but decreasing overall event frequency. Research with annual species in arid and semiarid ecosystems has demonstrated that precipitation variability can influence plant community dynamics; however, less is known about the impact of precipitation variability in less water-limited ecosystems, including economically important agricultural systems. * METHODS: We conducted three greenhouse experiments to determine how variation in total precipitation and the interval between precipitation events affected emergence and growth of two common annual midwestern weed species, Chenopodium album (Chenopodiaceae) and Setaria faberi (Poaceae). * KEY RESULTS: Both species responded to precipitation variability; however, the effect depended on life stage and precipitation amount, indicating that responses are highly context-dependent. Emergence of both species increased with longer intervals between precipitation events at low total precipitation, but species' responses varied under typical precipitation amounts. Individual seedling biomass of both species depended on interactions between total water and intervals, but species' responses differed; Setaria faberi biomass was reduced with longer intervals, but Chenopodium album had either a positive or no response. * CONCLUSIONS: Our results suggest that changes in precipitation variability likely will affect the composition and relative abundance of agriculturally important weeds. These results are important for understanding how changes in the temporal variability of precipitation due to global climate changes could impact plants in non-arid communities. PMID- 21616799 TI - Evolution of breeding systems and fruits in New World Galium and relatives (Rubiaceae). AB - PREMISE OF THE STUDY: Dioecy occurs in only about 6% of angiosperms, yet it has evolved many times from hermaphroditism. Polygamy is an even more uncommon condition within angiosperms, in which both unisexual and bisexual flowers occur within a species. Polygamy, dioecy, and hermaphroditism all occur within a New World clade of Galium (Rubiaceae), in which dioecy is hypothesized to have evolved from hermaphroditism via polygamy. At least five sections of Galium as traditionally defined by fruit morphology occur within this group. We tested the monophyly of sections defined by fruit morphology and sought to determine origins and pathways of breeding systems within this group. * METHODS: We obtained chloroplast (rpoB-trnC, trnC-psbM, trnL-ndhJ) and nuclear ribosomal (external transcribed spacer) DNA sequences for 89 taxa from the Cruciata-Galium-Valantia (CGV) clade to estimate the phylogeny. Ancestral states for breeding systems, fruit types, and fruit hairs were reconstructed using parsimony and likelihood analyses. * KEY RESULTS: We identified nine well-supported lineages of New World Galium taxa. However, none of the sections traditionally defined by fruit morphology are monophyletic. Dioecy is inferred to have arisen at least three times from hermaphroditism; polygamy is inferred to have arisen at least twice from dioecy and at least six times from hermaphroditism. * CONCLUSIONS: Polygamy appears to be a terminal condition in the CGV clade and not a pathway to dioecy. Fruit characters traditionally used in the taxonomy of this group have arisen multiple times within this clade of Galium and are not reliable indicators of shared evolutionary history. PMID- 21616800 TI - A molecular phylogeny and classification of Verbenaceae. AB - PREMISE OF THE STUDY: Verbenaceae consist of trees, shrubs, lianas, and herbs distributed primarily in Latin America, where they occur in a wide array of ecosystems. A second center of diversity exists in Africa. Competing morphology based classifications that rely on different traits conflict in significant ways. A broad phylogenetic study was undertaken to assess those classifications and to examine the historical geography of the family. * METHODS: Analysis of seven chloroplast DNA regions for 109 species, representing all genera except one monotypic genus, provide inference into evolutionary relationships in Verbenaceae. * KEY RESULTS: The phylogeny shows that none of the traditional classifications reflect phylogenetic relationships very well. Eight clades are recognized as tribes (Casselieae, Citharexyleae, Duranteae, Lantaneae, Neospartoneae trib. nov., Petreeae, Priveae, and Verbeneae). Two genera, Dipyrena and Rhaphithamnus, remain unplaced in these larger clades. Petreeae, which consist of Neotropical lianas, are sister to the rest of the family. Lantaneae and Verbeneae together form a derived clade that comprises approximately two thirds of the species in Verbenaceae. * CONCLUSIONS: We present a new tribal classification, including one new tribe, Neospartoneae trib. nov., to accommodate three small genera of Argentine species (Diostea, Neosparton, and Lampaya). Phylogenetic inference suggests a South American origin for Verbenaceae, with approximately six colonization events having given rise to the Old World species. PMID- 21616801 TI - Molecular phylogeny of Atripliceae (Chenopodioideae, Chenopodiaceae): Implications for systematics, biogeography, flower and fruit evolution, and the origin of C4 photosynthesis. AB - PREMISE OF THE STUDY: Atripliceae (Chenopodiaceae), including Atriplex (300 spp.) as the largest genus of the family, are an ecologically important group of steppes and semideserts worldwide. Relationships in Atripliceae are poorly understood due to obscure and potentially convergent morphological characters. * METHODS: Using sequence variation of two chloroplast markers (rbcL gene, atpB rbcL spacer) and one nrDNA marker (ITS) analyzed with BEAST, we investigated the systematics and biogeography of Atripliceae. We surveyed flower morphology and fruit anatomy to study the evolution of flowers and fruits in the tribe. * KEY RESULTS: Female flowers with persistent foliar cover (the diagnostic character of traditional Atripliceae) evolved three times in Chenopodioideae, in Atripliceae s.s., Axyrideae, and Spinacia. Atripliceae s.s. started to diversify during the Early Miocene in Eurasia, separating into the Archiatriplex and the Atriplex clades. The former consists of eight species-poor, disjunct, and morphologically heterogeneous genera and is likely a relictual lineage. The Atriplex clade comprises the majority of species and evolved one C(4) lineage 14.1-10.5 Ma, which diversified rapidly worldwide. The C(4) Atriplex entered North America during the Middle/Late Miocene and spread to South America subsequently. Australia was colonized by two C(4) lineages both arriving during the Late Miocene. One of them diversified rapidly, giving rise to most Australian Atriplex species. * CONCLUSIONS: Atripliceae s.s. comprise Archiatriplex, Atriplex, Exomis, Extriplex, Grayia, Halimione, Holmbergia, Manochlamys, Proatriplex, and Stutzia. Microgynoecium is included based on morphology but only weak molecular support. Axyris, Krascheninnikovia, and Ceratocarpus (here described as Axyrideae) and Spinacia are excluded from Atripliceae. PMID- 21616802 TI - Complex patterns of hybridization between exotic and native North American poplar species. AB - PREMISE OF THE STUDY: Poplars and their hybrids are seen as important candidates for bioenergy initiatives. However, many concerns have been raised about large scale plantations of new poplar cultivars. The deployment of such plants with novel traits brings the risk of potential spread of novel genome regions (including exotic genes, transgenes, or other heritable modifications) into natural populations of related species. The possibility of introgression is especially high in poplars because reproductive barriers between species are weak. Knowledge of the frequency of hybridization between cultivated trees and natural populations is one important step in the risk-assessment process. * METHODS: We studied the rate of spontaneous hybridization from two sexually mature poplar plantations into adjacent natural populations of Populus deltoides and P. balsamifera. The two plantations, both in eastern Canada, contain many different complex hybrid clones with components from exotic species, mostly P. nigra, P. trichocarpa, and P. maximowiczii. We analyzed 12 species-specific single nucleotide polymorphisms from six different genes in 5373 offspring sampled from the natural populations. * RESULTS: Contributions from all three exotics were found in the offspring, confirming low reproductive barriers among poplar species in these sections. The frequency of hybrid offspring varied among pollen donors, recipient populations, and years. * CONCLUSIONS: The remarkably high rate of hybridization that was found in the smallest natural population sampled suggests that small peripheral populations carry a higher risk of introgression. These results could be used as a starting point for developing regulatory guidelines for the introduction of plants with novel traits. PMID- 21616803 TI - Reproductive success of cultivated Pyrus calleryana (Rosaceae) and establishment ability of invasive, hybrid progeny. AB - * PREMISE OF THE STUDY: Pyrus calleryana Decne., an ornamental tree species introduced from China, is a relatively new invasive that has only recently begun to spread across the United States after intraspecific hybridization between cultivars. The function of such hybridization in the evolution of invasiveness is still relatively understudied, especially with respect to the initial establishment and persistence of invasive genotypes. Multiple introductions of genetically divergent populations or cultivars may benefit from new genetic combinations created during hybridization events and/or release from Allee effects in founder populations. * METHODS: We quantified the outcome of intraspecific hybridization between cultivars of P. calleryana in a common garden. Measures of the reproductive success and establishment ability of their early- and advanced-generation hybrid offspring were collected to assess the likelihood of particular cultivar genotypes to establish in invasive populations. These traits also were compared between cultivated and invasive parents to identify any generational differences in invasive potential. * KEY RESULTS: Differences were detected in measures of reproductive ability, but no group emerged as consistently more fecund. Advanced-generation hybrids also had significantly less biomass, indicating a reduction in hybrid performance relative to that of the cultivated progeny. * CONCLUSION: Ultimately, this study indicates that increased spread of P. calleryana has been initiated by introduction of multiple cultivar types and subsequent widespread planting and is not the result of an inherent fitness advantage of hybrid progeny. PMID- 21616804 TI - Morphologically cryptic biological species within the liverwort Frullania asagrayana. AB - PREMISE OF THE STUDY: The Frullania tamarisci complex includes eight Holarctic liverwort species. One of these, F. asagrayana, is distributed broadly throughout eastern North America from Canada to the Gulf Coast. Preliminary genetic data suggested that the species includes two groups of populations. This study was designed to test whether the two groups are reproductively isolated biological species. * METHODS: Eighty-eight samples from across the range of F. asagrayana, plus 73 samples from one population, were genotyped for 13 microsatellite loci. Sequences for two plastid loci and nrITS were obtained from 13 accessions. Genetic data were analyzed using coalescent models and Bayesian inference. * KEY RESULTS: Frullania asagrayana is sequence-invariant at the two plastid loci and ITS2, but two clear groups were resolved by microsatellites. The two groups are largely reproductively isolated, but there is a low level of gene flow from the southern to the northern group. No gene flow was detected in the other direction. A local population was heterogeneous but displayed strong genetic structure. * CONCLUSIONS: The genetic structure of F. asagrayana in eastern North America reflects morphologically cryptic differentiation between reproductively isolated groups of populations, near-panmixis within groups, and clonal propagation at local scales. Reproductive isolation between groups that are invariant at the level of nucleotide sequences shows that caution must be exercised in making taxonomic and evolutionary inferences from reciprocal monophyly (or lack thereof) between putative species. PMID- 21616805 TI - Quantitative variation for apomictic reproduction in the genus Boechera (Brassicaceae). AB - PREMISE OF THE STUDY: The evolution of asexual seed production (apomixis) from sexual relatives is a great enigma of plant biology. The genus Boechera is ideal for studying apomixis because of its close relation to Arabidopsis and the occurrence of sexual and apomictic species at low ploidy levels (diploid and triploid). Apomixis is characterized by three components: unreduced embryo-sac formation (apomeiosis), fertilization-independent embryogenesis (parthenogenesis), and functional endosperm formation (pseudogamy or autonomous endosperm formation). Understanding the variation in these traits within and between species has been hindered by the laborious histological analyses required to analyze large numbers of samples. * METHODS: To quantify variability for the different components of apomictic seed development, we developed a high throughput flow cytometric seed screen technique to measure embryo:endosperm ploidy in over 22000 single seeds derived from 71 accessions of diploid and triploid Boechera. * KEY RESULTS: Three interrelated features were identified within and among Boechera species: (1) variation for most traits associated with apomictic seed formation, (2) three levels of apomeiosis expression (low, high, obligate), and (3) correlations between apomeiosis and parthenogenesis/pseudogamy. * CONCLUSIONS: The data presented here provide a framework for choosing specific genotypes for correlations with large "omics" data sets being collected for Boechera to study population structure, gene flow, and evolution of specific traits. We hypothesize that low levels of apomeiosis represent an ancestral condition of Boechera, whereas high apomeiosis levels may have been induced by global gene regulatory changes associated with hybridization. PMID- 21616806 TI - Phylogeny and a new tribal classification of the Panicoideae s.l. (Poaceae) based on plastid and nuclear sequence data and structural data. AB - PREMISE OF THE STUDY: The subfamily Panicoideae (Poaceae) encompasses nearly one third of the diversity of grass species, including important crops such as maize and sugarcane. Previous analyses recovered strong support for a Panicoideae+Centothecoideae lineage within the diverse Panicoideae+Arundinoideae+Chloridoideae+Micrairoideae+Aristidoideae+Danthonioidea (PACMAD) clade, although support for internal relationships was inconsistent. The objectives of this research were to (1) further test the monophyly of each subfamily and previously recovered clades within the Panicoideae+Centothecoideae lineage, (2) establish phylogenetic relationships among these groups, and (3) propose a new tribal classification for this lineage based explicitly on the phylogeny. * METHODS: Maximum parsimony and Bayesian inference analyses of 37 taxa were based on previously published sequences (ndhF and rpl16 intron) and on new plastid and nuclear (rbcL and granule-bound starch synthase I) sequence data as well as structural data. * Key results. The Panicoideae+Centothecoideae lineage and a majority of the clades identified in previous analyses continue to be robustly supported, but resolution along the backbone of the topology remains elusive. Support for the monophyly of both subfamilies was lacking although support values for some clades increased. The tribes Centotheceae and Arundinelleae were confirmed as polyphyletic. * CONCLUSIONS: Subfamily Centothecoideae is formally submerged into the Panicoideae, and a new tribal classification for the expanded Panicoideae is proposed based explicitly on the phylogeny. This classification includes 12 tribes of which Chasmanthieae and Zeugiteae are segretated from the Centotheceae; Tristachyideae is segregated from Arundinelleae, and a new tribe, Cyperochloeae, is validated to accommodate two isolated genera. A key to the tribes is provided. PMID- 21616807 TI - Unusual patterns of hybridization involving a narrow endemic Rhododendron species (Ericaceae) in Yunnan, China. AB - PREMISE OF THE STUDY: One potential threat to rare species is genetic swamping caused by hybridization, but few studies have quantified this threat. Rhododendron cyanocarpum is a narrow endemic species that occurs sympatrically with potentially interfertile congeners throughout its range within Yunnan, China. We searched the entire distribution of R. cyanocarpum for hybrids and examined the patterns of hybridization to assess potential threat from hybridization. * METHODS: In a comprehensive field survey, we detected only one instance of hybridization involving R. cyanocarpum, with R. delavayi, at Huadianba near Dali. Material of both species and putative hybrids was examined using morphology, chloroplast DNA, nuclear ribosomal DNA, and Bayesian analysis of AFLP profiles. * KEY RESULTS: Of 10 putative hybrids, two were F(1)(')s and at least seven were F(2)(')s. Four backcrosses to R. delavayi were detected among material with R. delavayi-like morphology within the hybrid zone. Backcrosses to R. cyanocarpum were not detected. Therefore F(2)(')s outnumbered all other classes within the hybrid zone, a situation not previously confirmed for plants and extremely rare generally. Hybridization was asymmetrical, with R. delavayi as the maternal parent in all but one of the hybrids detected. * CONCLUSIONS: Although natural hybridization is common in Rhododendron, it is rare in R. cyanocarpum and is apparently not accompanied by backcrossing toward R. cyanocarpum. Hence, there is no immediate risk of genetic swamping, unless habitat disturbance increases and changes the patterns of hybridization. Our study is the first to report a plant hybrid zone dominated by F(2) hybrids. This pattern might contribute to species barrier maintenance. PMID- 21616808 TI - Characterization of 14 microsatellite loci in a tropical palm, Attalea phalerata (Arecaceae). AB - PREMISE OF THE STUDY: We developed microsatellite primers for the widely distributed tropical palm Attalea phalerata for studies on the dispersal and spatial genetic structure of palm populations. * METHODS AND RESULTS: Fourteen di , tri-, and tetra-nucleotide microsatellite primer pairs were identified. The number of alleles in the population tested ranged between 3 and 25, with a mean of 12.1. Ten microsatellite loci exhibited no significant deviations from Hardy Weinberg Equilibrium or presence of null alleles, and their combined probability of exclusion was 0.998. * CONCLUSIONS: These microsatellite loci will be useful in parentage analysis and population genetics studies of Attalea phalerata. PMID- 21616809 TI - Microsatellite loci for Paspalum atratum (Poaceae) and cross-amplification in other species. AB - PREMISE OF THE STUDY: Paspalum atratum is a perennial, cespitose, tropical grass native to Central and South America. This species belongs to a polyploid complex (Plicatula group) little known at the genetic level. The characterized microsatellite markers provide new informative tools for further studies of the hybridization, mating systems, and structure of the population. * METHODS AND RESULTS: Using the microsatellite-enriched library method, we isolated and characterized 19 microsatellite markers from P. atratum. Eleven of them were polymorphic, showing a variable degree of variation, while eight were monomorphic in the samples analyzed. Additionally, the transferability of these microsatellite markers was tested in other species. * CONCLUSIONS: These results suggest that the characterized markers have enough discriminatory potential to be used in genetic characterizations of Paspalum taxa, which are based on an understanding of their mating systems and genetic structure, as well as in understanding the evolutionary processes involved in the evolution of groups of Paspalum. PMID- 21616810 TI - Development and characterization of microsatellite loci for Khaya senegalensis (Meliaceae)1. AB - PREMISE OF THE STUDY: Microsatellite loci were developed to characterize genetic variation and population subdivision in Khaya senegalensis (Desr.) A. Juss. (Meliaceae). * METHODS AND RESULTS: Microsatellite loci were identified from genomic DNA sequences generated using the 454 GS-FLX titanium platform. Primers were designed for 67 tri- and tetranucleotide repeats, of which 20 were selected for 2 multiplexes based on amplification success and band size. Eleven of these loci showed polymorphism in two populations of Khaya senegalensis and amplified in individuals from across the species range. * CONCLUSIONS: These new microsatellite loci will be useful for investigation of the evolutionary and conservation genetics of Khaya senegalensis. PMID- 21616811 TI - Development of microsatellite loci for the invasive weed Wedelia trilobata (Asteraceae). AB - PREMISE OF THE STUDY: Microsatellite markers were developed to help elucidate the population genetics of the invasive species Wedelia trilobata. * METHODS AND RESULTS: Using the Fast Isolation by AFLP of Sequences COntaining (FIASCO) repeats protocol, 23 sets of primers for amplifying microsatellite loci were identified in W. trilobata, 10 of which showed polymorphism (two to five alleles per locus) in samples of two populations of W. trilobata, one from China and one from Peru. Six of these loci were successfully amplified from samples of the native congener W. chinensis, with expected sizes. * CONCLUSIONS: These markers may be useful for further investigation of population genetics of Wedelia trilobata and other congener species. PMID- 21616812 TI - Isolation and characterization of 50 nuclear microsatellite markers for Cathaya argyrophylla, a Chinese endemic conifer. AB - PREMISE OF THE STUDY: Microsatellite primers were developed for the endangered Cathaya argyrophylla (Pinaceae) to investigate its genetic diversity and population genetic structure, as well as its evolutionary history. * METHODS AND RESULTS: Fifty dinucleotide microsatellite loci were identified in two populations. The number of alleles per locus ranged from 1 to 6, with a mean of 2.84. The observed and expected heterozygosities ranged from 0 to 0.889 and from 0 to 0.779, respectively. * CONCLUSIONS: These markers will facilitate further studies on the population genetics and evolutionary history of Cathaya argyrophylla. PMID- 21616813 TI - New microsatellite loci in the dwarf yams Dioscorea group Epipetrum (Dioscoreaceae). AB - PREMISE OF THE STUDY: Microsatellite loci were isolated and characterized from enriched genomic libraries of two taxa of the Chilean Epipetrum group of Dioscorea to assess their levels of genetic diversity and population differentiation. * METHODS AND RESULTS: Eleven microsatellite loci were identified. Six out of nine microsatellites from D. biloba amplified in D. humilis, and the two microsatellites from D. humilis amplified in both taxa. Two different sets of eight loci amplified in each of the two tested taxa, D. biloba and D. humilis. The average number of alleles was 5.75 and 5 for D. biloba and D. humilis, respectively. Higher levels of mean genetic diversity were found in D. biloba (H(E) = 0.639) than in D. humilis (H(E) = 0.414). * CONCLUSIONS: These microsatellite primers will be useful in population genetic studies and to establish conservation strategies in the endangered taxa of the Epipetrum group of Dioscorea. PMID- 21616814 TI - Development of SSR markers from Citrus clementina (Rutaceae) BAC end sequences and interspecific transferability in Citrus. AB - PREMISE OF THE STUDY: Microsatellite primers were developed from bacterial artificial chromosome (BAC) end sequences of Citrus clementina and their transferability and polymorphism tested in the genus Citrus for future anchorage of physical and genetic maps and comparative interspecific genetic mapping. * METHODS AND RESULTS: Using PAGE and DNA silver staining, 79 primer pairs were selected for their transferability and polymorphism among 526 microsatellites mined in BES. A preliminary diversity study in Citrus was conducted with 18 of them, in C. reticulata, C. maxima, C. medica, C. sinensis, C. aurantium, C. paradisi, C. lemon, C. aurantifolia, and some papedas (wild citrus), using a capillary electrophoresis fragment analyzer. Intra- and interspecific polymorphism was observed, and heterozygous markers were identified for the different genotypes to be used for genetic mapping. * CONCLUSIONS: These results indicate the utility of the developed primers for comparative mapping studies and the integration of physical and genetic maps. PMID- 21616815 TI - Modeling Nymphoides architecture: A morphological analysis of Nymphoides aquatica (Menyanthaceae). AB - PREMISE OF THE STUDY: Species in the aquatic genus Nymphoides have inflorescences that appear to arise from the petioles of floating leaves. The inflorescence-floating leaf complex can produce vegetative propagules and/or additional inflorescences and leaves. We analyzed the morphology of N. aquatica to determine how this complex relates to whole plant architecture and whether whole plant growth is sympodial or monopodial. * METHODS: We used dissections, measurements, and microscopic observations of field-collected plants and plants cultivated for 2 years in outdoor tanks in south Florida, USA. * KEY RESULTS: Nymphoides aquatica had a submerged plagiotropic rhizome that produced floating leaves in an alternate/spiral phyllotaxy. Rhizomes were composed of successive sympodial units that varied in the number of leaves produced before the apex terminated. The basic sympodial unit had a prophyll that subtended a renewal shoot bud, a short-petioled leaf (SPL) with floating lamina, and an inflorescence; the SPL axillary bud expanded as a vegetative propagule. Plants produced either successive basic sympodial units or expanded sympodia that intercalated long-petioled leaves between the prophyll and the SPL. * CONCLUSIONS: Nymphoides aquatica grows sympodially, forming a rhizome composed of successive basic sympodia and expanded sympodial units. Variations on these types of sympodial growth help explain the branching patterns and leaf morphologies described for other Nymphoides species. Monitoring how these two sympodial phases are affected by water depth provides an ecologically meaningful way to assess N. aquatica's responses to altered hydrology. PMID- 21616816 TI - Resource use efficiency and community effects of invasive Hypochaeris radicata (Asteraceae) during primary succession. AB - PREMISE OF THE STUDY: We sought to better understand the impacts and mechanisms underpinning a successful invasion of resource-poor sites by a nonnative plant on Mount St. Helens volcano (MSH). * METHODS: We investigated the short-term effects of the nonnative plant Hypochaeris radicata on growth of native species colonizing drought-prone primary successional surfaces under N-limited and N augmented conditions. To understand the success of H. radicata, we compared its resource use efficiency to that of a closely related native colonist, Hieracium albiflorum, under the same conditions. * KEY RESULTS: Removing H. radicata did not affect growth of the most common colonists, but N addition demonstrated strong N limitation to growth in H. albiflorum, H. radicata, and Agrostis spp. Nonnative H. radicata exhibited lower water-use efficiency than H. albiflorum but did not differ in efficiency of N use. H. radicata biomass increased faster in response to an N pulse than did the native H. albiflorum, as did the pool of N held in H. radicata tissues. * CONCLUSIONS: Our findings contrast with results from Hawaiian volcanic sites, where higher short-term resource use efficiency was reported for invasive species, including H. radicata. Our results suggest that at MSH, the success of H. radicata relies on rapid uptake and utilization of N rather than on higher efficiency. This strategy is especially advantageous at MSH because N pulses commonly occur as a consequence of herbivore-induced mortality of Lupinus lepidus (Fabaceae). PMID- 21616817 TI - Seed ecology of an invasive alien species, Acacia longifolia (Fabaceae), in Portuguese dune ecosystems. AB - PREMISE OF THE STUDY: Worldwide, invasive plants threaten biodiversity, by disrupting habitats and ecosystem processes, and cause major economic losses. Invasiveness in plants is frequently associated with prolific production of seeds that accumulate in the soil. Knowledge of the extent and persistence of invasive seed banks helps explain invasion processes and enables management planning. A study of Acacia longifolia, an invasive species in Portuguese dune ecosystems, provides an informative example. * METHODS: Seed rain and dispersal (seed traps), the persistence of seeds in the soil (burial), and the extent of seed banks were measured and analyzed. * KEY RESULTS: Seed rain is concentrated under the canopy with about 12000 seeds . m(-2) falling annually. The number of seeds in the soil declined with time, with only 30% surviving after 75 mo. Losses were lowest at greater depths. Seed germinability was low (<12%), but viability was high (>85%) for surviving seeds. The seed bank under the canopy was approximately 1500 and 500 seeds . m(-2) in long- and recently invaded stands, respectively. Some seeds were found up to 7 m from the edge of stands, indicating that outside agencies facilitate dispersal. * CONCLUSIONS: Acacia longifolia produces large numbers of seeds, some of which are lost through germination, decay, and granivory. The remainder form vast and persistent seed banks that serve as a source of replenishment and make it difficult to control the invader once it is established. Control costs escalate as the duration of an invasion increases, highlighting the urgency of initiating and persevering with control efforts. PMID- 21616818 TI - Seedling dynamics at elevation limits: Complex interactions beyond seed and microsite limitations. AB - PREMISE OF THE STUDY: Limited availability of seeds and microsites are important constraints for seedling emergence and survival. However, little is known about population-level feedbacks between seed and microsite limitation and how these relationships vary in contrasting conditions. Focusing on Armeria caespitosa, a high-mountain endemic, we asked whether seedling establishment was simultaneously limited by seed and microsite availability, whether the balance between seed and microsite limitations varied with contrasting environments, and whether seed and microsite limitations interacted with each other. * METHODS: We studied seedling emergence and survival at the edges of the elevation range of A. caespitosa in central Spain over 4 years. We used mixed linear models and structural equation modeling. * KEY RESULTS: Our results showed that A. caespitosa recruits similar numbers of new seedlings per plot in contrasting environments. Seedling emergence and survival in A. caespitosa were both limited by seed and microsite availability. Habitat suitability (the surrogate of microsite limitation) varied between populations and indirectly affected fertility (the surrogate of seed limitation). * CONCLUSIONS: Seedling emergence and survival are both seed and microsite limited, there are interactions between seed and microsite limitations, and both factors, which control seedling establishment, and their interconnections, vary among local populations. PMID- 21616819 TI - Ploidy-specific interactions of three host plants with arbuscular mycorrhizal fungi: Does genome copy number matter? AB - PREMISE OF THE STUDY: Polyploidy has been shown to affect different plant traits and modulate interactions between plants and other organisms, such as pollinators and herbivores. However, no information is available on whether it can also shape the functioning of mycorrhizal symbiosis. * METHODS: The mycorrhizal growth response was assessed for three angiosperms with intraspecific ploidy variation. Different cytotypes of Aster amellus, Campanula gentilis, and Pimpinella saxifraga were either left uninoculated or were inoculated with arbuscular mycorrhizal (AM) fungi in a pot experiment. After 3 mo of cultivation in a greenhouse, plant growth, phosphorus concentration in the shoot biomass, and development of the AM symbiosis were evaluated. * KEY RESULTS: No significant ploidy-specific differences in AM development were recorded. The inoculation led to consistently greater phosphorus uptake; however, the effect on plant growth differed considerably among plant species, populations, ploidy levels, and AM species. A salient ploidy-specific response was observed in A. amellus. Whereas diploid plants benefited from AM inoculation, the hexaploids consistently showed negative or no-growth responses (depending on the AM species). In contrast to A. amellus, no interactions between inoculation and ploidy were observed in C. gentilis and P. saxifraga. * CONCLUSIONS: The first evidence is provided of a ploidy-specific response of a mycotrophic plant to AM fungi. Our results demonstrate the complexity of interaction between plants and associated AM fungi, with the ploidy level of the host plant being one component that may modulate the functioning of the symbiosis. PMID- 21616820 TI - Zero-sum allocational strategies determine the allometry of specific leaf area. AB - PREMISE OF THE STUDY: Specific leaf area (SLA) is a critical component of the leaf economics spectrum, and many functional leaf traits have been empirically demonstrated to covary with SLA. However, a complete understanding of how change in leaf size influences SLA has not yet emerged. * METHODS: To help develop a more complete understanding of the determinants of variability in SLA, we present a covariation model of leaf allometry that predicts a zero-sum interdependence of leaf thickness, density, and surface area on leaf mass. We test the model's predictions on measurements of 900 leaves from 44 angiosperm species. * KEY RESULTS: We observe that "diminishing returns," the negative allometry (slope < 1) of surface area versus mass, does not hold universally across species. Rather, the scaling of SLA is linked to the relative allocation to thickness and density. Specifically, diminishing returns are observed when leaves grow thicker, more than their density decreases, with increasing mass. Finally, we confirm model predictions that the allometric dependence of area, thickness, and density on mass can be well approximated by a zero-sum allocational process. * CONCLUSIONS: Our work adds to the growing body of evidence that allometric covariation is a hallmark of the scaling behavior of complex plant and leaf traits. Moreover, because our model makes predictions based on allocational constraints, it provides a foundation to understand how deviations from zero-sum tradeoffs in allocation to leaf thickness, density, or area determine the allometry of SLA and, ultimately, underlie adaptive strategies within and across plant species. PMID- 21616821 TI - Morphometric architecture of the most taxon-rich species in the U.S. flora: Astragalus lentiginosus (Fabaceae). AB - PREMISE OF THE STUDY: The study of infra-taxa has historically been considered the study of incipient species. Astragalus lentiginosus Douglas ex Hooker (Fabaceae) is the most taxonomically diverse species in the U. S. flora. To gain insight into the nature of infra-taxa, I ask whether an optimal number of groups exists within A. lentiginosus, whether the varieties represent discrete entities, and whether morphological diversity within A. lentiginosus can be explained by climate. * METHODS: Morphometric measurement was made on herbarium specimens. Seasonality was inferred by fitting sine waves to climatic data. Statistical clustering, ordination, and multiple regression analyses were employed to test hypotheses. * KEY RESULTS: K-means clustering resulted in no clear optimal number of groups within the species. Principal components analysis demonstrated that taxa occupy contiguous regions of morphospace but do not form distinct clusters. Multiple regression demonstrated significant relations among morphology and climatic parameters, which are generally stronger than geographic relations. * CONCLUSIONS: The bewildering amount of diversity contained within A. lentiginosus begs for decomposition, yet its clinal nature precludes it from division into discrete groups. As a species, A. lentiginosus exists as an array of populations in a delicate balance between homogenizing forces, such as gene flow and retention of ancestral characters, and diversifying forces, such as local adaptation and drift. The effect of these forces has become evident in the phenotype of A. lentiginosus, resulting in more infra-taxa than any other species in the U. S. flora, but has failed to fracture this group into distinct entities. PMID- 21616822 TI - Molecular phylogenetics of suborder Cactineae (Caryophyllales), including insights into photosynthetic diversification and historical biogeography. AB - PREMISE OF THE STUDY: Phylogenetic relationships were investigated among the eight families (Anacampserotaceae, Basellaceae, Cactaceae, Didiereaceae, Halophytaceae, Montiaceae, Portulacaceae, Talinaceae) that form suborder Cactineae (= Portulacineae) of the Caryophyllales. In addition, photosynthesis diversification and historical biogeography were addressed. * METHODS: Chloroplast DNA sequences, mostly noncoding, were used to estimate the phylogeny. Divergence times were calibrated using two Hawaiian Portulaca species, due to the lack of an unequivocal fossil record for Cactineae. Photosynthetic pathways were determined from carbon isotope ratios (delta(13)C) and leaf anatomy. * KEY RESULTS: Maximum likelihood and Bayesian analyses were consistent with previous studies in that the suborder, almost all families, and the ACPT clade (Anacampserotaceae, Cactaceae, Portulacaceae, Talinaceae) were strongly supported as monophyletic; however, relationships among families remain uncertain. The age of Cactineae was estimated to be 18.8 Myr. Leaf anatomy and delta(13)C and were congruent in most cases, and inconsistencies between these pointed to photosynthetic intermediates. Reconstruction of photosynthesis diversification showed C(3) to be the ancestral pathway, a shift to C(4) in Portulacaceae, and five independent origins of Crassulacean acid metabolism (CAM). Cactineae were inferred to have originated in the New World. * CONCLUSIONS: Although the C(3) pathway is inferred as the ancestral state in Cactineae, some CAM activity has been reported in the literature in almost every family of the suborder, leaving open the possibility that CAM may have one origin in the group. Incongruence among loci could be due to internal short branches, which possibly represent rapid radiations in response to increasing aridity in the Miocene. PMID- 21616823 TI - Multilocus patterns of nucleotide polymorphism and demographic change in Taxodium distichum (Cupressaceae) in the lower Mississippi River alluvial valley. AB - PREMISE OF THE STUDY: Studies of the geographic patterns of genetic variation can give important insights into the past population structure of species. Our study species, Taxodium distichum L. (bald-cypress), prefers riparian and wetland habitats and is widely distributed in southeastern North America and Mexico. We compared the genetic variation of T. distichum with that of its close relative, Cryptomeria japonica, which is endemic to Japan. * METHODS: Nucleotide polymorphisms of T. distichum in the lower Mississippi River alluvial valley, USA, were examined at 10 nuclear loci. * KEY RESULTS: The average nucleotide diversity at silent sites, pi(sil), across the 10 loci in T. distichum was higher than that of C. japonica (pi(sil) = 0.00732 and 0.00322, respectively). In T. distichum, Tajima's D values were each negative at 9 out of 10 loci, which suggests a recent population expansion. Maximum-likelihood and Bayesian estimations of the exponential population growth rate (g) of T. distichum populations indicated that this species had expanded approximately at the rate of 1.7-1.0 * 10(-6) per year in the past. * CONCLUSIONS: Taxodium distichum had significantly higher nucleotide variation than C. japonica, and its patterns of polymorphism contrasted strikingly with those of the latter, which previously has been inferred to have experienced a reduction in population size. PMID- 21616824 TI - Other tetraploid species and conspecific diploids as sources of genetic variation for an autotetraploid. AB - PREMISE OF THE STUDY: Most plants are polyploid and have more than two copies of the genome. The evolutionary success of polyploids is often attributed to their potential to harbor increased genetic variation, but it is poorly understood how polyploids can attain such variation. Because of their formation bottleneck, newly formed tetraploids start out with little variation. Tetraploids may attain genetic variation through a combination of new mutations, recurrent formation, and gene exchange with diploid ancestors or related tetraploid species. We explore the role of gene exchange and introgression in autotetraploid Rorippa amphibia, a species that harbors more genetic variation than its diploid ancestors. * METHODS: We crossed autotetraploid R. amphibia to diploid conspecifics and tetraploid R. sylvestris and backcrossed resulting F(1) hybrids. We used flow cytometry to determine the ploidy of all progeny. * KEY RESULTS: Tetraploids of R. amphibia and R. sylvestris were interfertile; F(1) hybrids were fertile and could backcross. Crosses between diploids and tetraploids yielded a small number of viable, often tetraploid progeny. This indicates that unreduced gametes can facilitate gene flow from diploids to tetraploids. We detected a frequency of unreduced gametes of around 2.7 per 1000, which was comparable between diploids and tetraploids. * CONCLUSIONS: Introgression from tetraploid R. sylvestris provides a realistic source of variation in autotetraploid R. amphibia. Only in a scenario where other compatible partners are absent, for example immediately after tetraploidization, gene flow through unreduced gametes from diploids could be an important source of genetic variation for tetraploids. PMID- 21616825 TI - Ectomycorrhizas of Cercocarpus ledifolius (Rosaceae). AB - PREMISE OF THE STUDY: Woody species in the Rosaceae form ectomycorrhizal associations, but the fungal symbionts are unknown. The species of fungi determine whether host plants are isolated from other ectomycorrhizal species in the plant community or linked with other trees through mycorrhizal networks. In this study we identified the fungi that form ectomycorrhizas with Cercocarpus ledifolius (curl-leaf mountain mahogany). * METHODS: Soil samples were collected under canopies of C. ledifolius. Ectomycorrhizas were described by morphology and by DNA sequences of the ITS region. Host species were confirmed by rbcL sequences. * KEY RESULTS: Sixteen species of fungi were identified from ectomycorrhizas of Cercocarpus ledifolius. The ectomycorrhizal community was distinguished by the presence of a Geopora species situated in the G. arenicola clade and by the absence of Rhizopogon, suilloids, and Sebacinales. Of the species on C. ledifolius, two also occurred on trees of Quercus garryana var. breweri and four on Arctostaphylos sp. * CONCLUSIONS: The presence of fungal species in common with other ectomycorrhizal hosts shows that C. ledifolius, Q. garryana var. breweri, and Arctostaphylos species could be linked by a mycorrhizal network, allowing them to exchange nutrients or to share inoculum for seedling roots and new fine roots. Single-host fungi limited to C. ledifolius may improve resource acquisition and reduce competition with other ectomycorrhizal hosts. The finding of a Geopora species as a frequent mycobiont of C. ledifolius suggests that this fungus might be appropriate for inoculating seedlings for habitat restoration. PMID- 21616826 TI - Organization, anatomy, and fungal endophytes of a Triassic conifer embryo. AB - PREMISE OF THE STUDY: Much is known about the Paleozoic conifers and the conifers assignable to extant families that appear in the Jurassic; however, relatively little is known about the transitional conifers of the early Mesozoic, especially the Voltziales. To better understand the evolution of this group, we aim to increase knowledge of voltzialean anatomy and morphology. A fossil embryo of the group is described in this study. * METHODS: Several permineralized seeds, one containing a well-preserved embryo, were collected from the early Middle Triassic Fremouw Peak locality in the Central Transantarctic Mountains. Samples were prepared using the standard acetate peel technique and studied in transmitted light. * KEY RESULTS: The embryo belongs to Parasciadopitys aequata, a member of the Voltziales. The embryo and megagametophyte tissues are exquisitely preserved. The embryo is colonized by two distinct fungi. Sporocarps of the fungi are found in the megagametophyte and in the space between the megagametophyte and nucellus. The additions of embryo and megagametophyte characters to the description of P. aequata have made it one of the most completely known fossil taxa reproductively. * CONCLUSIONS: Preservation of fossil embryos, although rare, can expand the array of characters available in tracing the evolutionary history of plants. The embryo of P. aequata shares similarities with embryos of other extinct and extant conifer families. The association of the embryo with Combresomyces cornifer and Mycocarpon asterineum increases our understanding of the roles of microorganisms in Triassic ecosystems. PMID- 21616827 TI - Characterization of oil and starch accumulation in tubers of Cyperus esculentus var. sativus (Cyperaceae): A novel model system to study oil reserves in nonseed tissues. AB - PREMISE OF THE STUDY: Storage oil (triacylglycerol) accumulates in tissues such as the embryo and endosperm of seeds and the fruit mesocarp, but seldom in underground organs. As a rare exception, cultivated variants of yellow nutsedge (Cyperus esculentus) contain high amounts of both oil and starch in the mature tubers. * METHODS: Biochemical analyses and light and electron microscopy were used to study the accumulation patterns of storage nutrients in developing nutsedge tubers. * KEY RESULTS: During the initial phase of tuber development, the conducting rhizome tissue is transformed into a storage compartment, then massive storage reserves accumulate in the tuber. At the beginning of tuber development, a large sugar load coincided with the onset of starch accumulation. Oil accumulation started later, concomitant with a substantial drop in the sugar content. Initially, oil accumulated at a lower rate compared to starch, but the rate later increased; after 6 wk, oil made up 24% of tuber dry mass, while starch made up 32%. Protein concentration changed only a small amount throughout this development. Oil and starch accumulated in the same cells throughout the tubers in a sequential fashion during tuber development. * CONCLUSIONS: The developmental pattern in the build up of storage nutrients in the tubers highlights nutsedge as a novel model plant, having potential to significantly widen our understanding on how synthesis of storage reserves, and in particular oils, is regulated and directed in nonseed tissues such as tubers and roots. PMID- 21616828 TI - Variation in the timing of autonomous selfing among populations that differ in flower size, time to reproductive maturity, and climate. AB - PREMISE OF THE STUDY: Early reproductive maturity is common in dry and ephemeral habitats and often associated with smaller flowers with increased potential for within-flower (autonomous) self-pollination. We investigated whether populations from locations that differ in moisture availability, known to vary for whole plant development rate, also varied in the timing of autonomous selfing. This timing is of interest because the modes of selfing (prior, competing, and delayed) have different fitness consequences. * METHODS: We measured timing of anther dehiscence, stigma receptivity, and herkogamy under pollinator-free conditions for plants from three populations of Collinsia parviflora that differed in annual precipitation, flower size, and time to sexual maturity. Using a manipulative experiment, we determined potential seed production via prior, competing, and delayed autonomous selfing for each population. * KEY RESULTS: Stigma receptivity, anther dehiscence, and selfing ability covaried with whole plant development and climate. Plants from the driest site, which reached sexual maturity earliest, had receptive stigmas and dehiscent anthers in bud. Most seeds were produced via prior selfing. The population from the wettest site with slowest development was not receptive until after flowers opened. Although competing selfing was possible, all selfing was delayed. The intermediate population was between these extremes, with significant contributions from both competing and delayed selfing. * CONCLUSIONS: Our results demonstrate that within species variation in the timing of selfing occurs and is related to both environmental conditions and whole-plant development rates. We suggest that, if these results can be generalized to other species, mating systems may evolve in response to ongoing climatic change. PMID- 21616829 TI - Phylogeographic structure of jack pine (Pinus banksiana; Pinaceae) supports the existence of a coastal glacial refugium in northeastern North America. AB - PREMISE OF THE STUDY: The genetic structure of jack pine (Pinus banksiana Lamb.), a North American boreal conifer with large longitudinal distribution, was investigated to test for the possible existence of a genetically distinct lineage in the Maritimes region in northeastern North America, which could be indicative of a mid-latitude coastal refuge during the last glaciation. * METHODS: One maternally inherited mitochondrial DNA (mtDNA) minisatellite marker and four paternally inherited chloroplast DNA (cpDNA) microsatellite markers were used to assess the range-wide geographical structure of jack pine populations with particular focus on northeastern North America. * KEY RESULTS: The populations from the Maritimes region presented a unique mtDNA background characterized by very low diversity and the preponderance of a distinctive mitotype. The distribution of cpDNA diversity was not spatially structured, though three chlorotypes were restricted to the east. * CONCLUSIONS: MtDNA data suggest that populations from the Maritimes region derive from a genetically depauperated north-coastal refugium. Contrastingly, the much higher geographical uniformity observed for cpDNA variation indicates that gene flow by pollen had been much more effective than seed gene flow at homogenizing population structure. PMID- 21616830 TI - Optimized genotyping with microsatellite markers in the fungal banana pathogen Mycosphaerella fijiensis (Mycosphaerellaceae). AB - PREMISE OF THE STUDY: Large-scale population genetics studies are required to investigate the dispersal processes underlying the emergence of Mycosphaerella fijiensis, a fungal pathogen of banana. To this end, we have developed an optimized genotyping procedure combining novel microsatellite markers and a modified DNA extraction protocol. * METHODS AND RESULTS: Primers for tetranucleotide loci were designed directly from the recently published genome sequence of M. fijiensis. A total of 19 new polymorphic and easy-to-score markers were developed. Their use was combined with an adapted protocol for total DNA extraction starting from young lesions collected from banana leaves, thus avoiding a pathogen isolation step. * CONCLUSIONS: The combination of the two technical developments presented here will permit the expansion of genotyping capacity in M. fijiensis, allowing large-scale analysis of samples from various geographic locations. PMID- 21616831 TI - De novo sequencing and development of EST-SSR for Gevuina avellana (Proteaceae), a nut tree from South America. AB - * PREMISE OF THE STUDY: The aim of this study was to develop molecular tools to investigate the genetic structure and diversity of natural populations of Gevuina avellana (Molina, Proteaceae), the Chilean hazelnut. Simple sequence repeats (SSRs) derived from expressed sequence tags (EST-SSRs) were developed, optimized, and characterized. * METHODS AND RESULTS: The microsatellite-containing sequences were selected from a cDNA library developed from the nut. The eight marker loci showed two to seven alleles in 60 unrelated trees, from two different natural populations. The observed heterozygosity (H(o)) was 0.44, ranging from 0.07 to 0.92 for different loci. When the multilocus genotype of the eight EST-SSRs was considered, all trees could be differentiated. * CONCLUSIONS: The newly identified EST-SSRs in G. avellana and the multi-pooling strategy involving eight markers will facilitate future studies of clone identification, genetic structure, and diversity. PMID- 21616832 TI - Development and characterization of microsatellite loci in Taihangia rupestris (Rosaceae), a rare cliff herb. AB - PREMISE OF THE STUDY: Microsatellite primers were developed for the rare Taihangia rupestris (Rosaceae) to evaluate genetic diversity, population genetic structure, mating system, and demographic events of this species. * METHODS AND RESULTS: Ten primer sets were developed using an enriched genomic library and were successfully amplified in T. rupestris var. ciliata and T. rupestris var. rupestris. The number of alleles per locus ranged from 2 to 21; the observed and expected heterozygosities ranged from 0.300 to 0.950 and from 0.328 to 0.956, respectively, in the two varieties. * CONCLUSIONS: The markers described here will be useful for studies of genetic variation, genetic structure, and mating systems of T. rupestris, which are important for the future conservation of this rare species. PMID- 21616833 TI - Isolation of compound microsatellite markers for the endangered plant Neolitsea sericea (Lauraceae). AB - PREMISE OF THE STUDY: The development of compound microsatellite markers was conducted in Neolitsea sericea to investigate genetic diversity and population genetic structure of this endangered insular species. * METHODS AND RESULTS: Using the compound microsatellite marker technique, 10 compound microsatellite markers that were successfully amplified showed polymorphism when assessed in 55 individuals from two populations in East China and Japan. Overall, the number of alleles ranged from 3 to 17, with an average of 7.9 alleles per locus. In addition, these primers could be easily amplified in Neolitsea aurata var. paraciculata and N. aurata var. chekiangensis. * CONCLUSIONS: The highly polymorphic markers developed and characterized in this study will be useful for population genetic studies of N. sericea. PMID- 21616834 TI - Development of intron targeting (IT) markers for potato and cross-species amplification in Solanum nigrum (Solanaceae). AB - PREMISE OF THE STUDY: Intron Targeting (IT) primers were developed for potato using expressed sequence tags (EST) and NCBI database records to study genetic diversity. * METHODS AND RESULTS: Twenty-nine polymorphic intron targeting (IT) markers were generated and characterized from 30 samples of potato and 22 samples of Solanum nigrum to detect polymorphism. The number of alleles (A) per locus ranged from 2 to 7 in the analyzed populations, and the observed heterozygosity (H(O)) and expected heterozygosity (H(E)) from 0 to 0.833 and 0.750, respectively. All of the primers also amplified in the related species S. nigrum. * CONCLUSIONS: The developed markers will provide valuable tools for genetic diversity analysis, genetic mapping, and marker-assisted breeding of potato and related Solanum species. PMID- 21616835 TI - New primers for single-copy nuclear-encoded chloroplast-expressed glutamine synthetase (ncpGS) in Oxalidaceae. AB - PREMISE OF THE STUDY: New primers were developed for the nuclear marker glutamine synthetase (ncpGS) in Oxalidaceae. * METHODS AND RESULTS: New forward and reverse primers were designed and tested across a wide range of Oxalidaceae. Selected taxa were sequenced to confirm homology. Potential for phylogenetic study was assessed by comparing sequenced taxa with commonly used nuclear and plastid markers. * CONCLUSIONS: Four out of five Oxalidaceae genera and all tested Oxalis spp. amplified successfully. Sequencing confirmed homology of the amplicon. Parsimony analysis of ncpGS showed that it is a promising candidate for future phylogenetic work in Oxalidaceae. PMID- 21616836 TI - Molecular genotyping of Trapa bispinosa and T. japonica (Trapaceae) based on nuclear AP2 and chloroplast DNA trnL-F region. AB - * PREMISE OF THE STUDY: Two marker systems were developed for the molecular identification of three Trapa species based on the length variation of nuclear AP2 and trnL-F chloroplast intergenic spacer region. * METHODS AND RESULTS: Our marker systems analyzed 118 individuals among 36 populations from three Trapa species. Trapa incisa was found to have a unique genotype from the other two species. Individuals of T. bispinosa were distinguished from T. japonica because the former had only a single AP2 genotype. Moreover, our results suggest a hybrid status of T. japonica in terms of two bands within an individual. One band was specific to T. japonica and the other one shared with T. bispinosa. * CONCLUSIONS: Our marker system demonstrates that nucleotide sequence variations can serve as a fast, reliable, and reproducible tool for molecular genotyping and examining the natural hybrid of water chestnut species. PMID- 21616837 TI - Identification and characterization of 74 novel polymorphic EST-SSR markers in the tea plant, Camellia sinensis (Theaceae). AB - PREMISE OF THE STUDY: New simple sequence repeat (SSR) markers were developed in the tea plant (Camellia sinensis) using published Expressed Sequence Tag (EST) sequences for further genetic studies and breeding programs. * METHODS AND RESULTS: A total of 74 EST-based SSR markers were generated. Polymorphism and transferability validation in 45 individuals of seven species and varieties of Camellia L. sect. Thea (L.) Dyer revealed that the number of alleles (N(A)) per locus varied between one and four or five in each species or variety. The observed heterozygosity (H(O)) and expected heterozygosity (H(E)) ranged from 0.000 to 1.000 and 0.893, respectively. * CONCLUSIONS: These new polymorphic and transferable EST-SSR markers will have potential for applications in genetic diversity evaluation, molecular fingerprinting identification, comparative genomics analysis, and genetic mapping in the tea plant. PMID- 21616838 TI - Ten polymorphic microsatellite markers in Michelia maudiae (Magnoliaceae). AB - PREMISE OF THE STUDY: Michelia maudiae is a threatened species in the Magnoliaceae. Microsatellite markers were developed and characterized in M. maudiae for further investigation of its conservation genetics. * METHODS AND RESULTS: Microsatellite markers were developed in M. maudiae using the Fast Isolation by AFLP of Sequences Containing Repeats protocol. Ten polymorphic microsatellites were assessed in two populations of M. maudiae. The number of alleles detected per locus varied from 1 to 8. Observed and expected heterozygosities ranged from 0.000 to 0.792 and from 0.000 to 0.826, respectively. Six primer pairs showed transferability in the two related species Michelia foveolata and Michelia chapensis. * CONCLUSIONS: This set of microsatellite markers provides a useful tool for future studies of the conservation genetics of M. maudiae and other congeneric species. PMID- 21616839 TI - Development of seven novel EST-SSR markers from Cycas panzhihuaensis (Cycadaceae). AB - PREMISE OF THE STUDY: Cycas panzhihuaensis L. Zhou & S. Y. Yang is a vulnerable gymnosperm endemic to China, where its range represents the northernmost occupation of Cycas L. We developed EST-derived SSR markers to investigate its genetic diversity and population structure. * METHODS AND RESULTS: Based on the expressed sequence tag (EST) database for Cycas rumphii Miq., seven simple sequence repeat (SSR) markers were identified and screened in 55 individuals from seven wild populations of C. panzhihuaensis. Alleles numbered 2 to 5, and their observed heterozygosity and expected heterozygosity ranged from 0.0000 to 0.6545 and from 0.0535 to 0.6966, respectively. * CONCLUSIONS: These new EST-SSR markers will enhance further studies of the population genetics of C. panzhihuaensis, allowing researchers to design reasonable conservation and management protocols. PMID- 21616840 TI - Immunohistochemical investigation of the necrotrophic phase of the fungus Colletotrichum gloeosporioides in the biocontrol of hemp sesbania (Sesbania exaltata; Papilionaceae). AB - PREMISE OF THE STUDY: Fungal plant pathogens exert much of their effect on plant cells through alterations in the host cell walls. However, obtaining biochemical proof for this change is difficult because of the relatively small number of cells that are affected by the pathogen relative to the bulk of host tissue. In this study, we examined the differences in host wall composition between infected and uninfected areas of seedlings of the weed hemp sesbania (Sesbania exaltata) that were treated with the biocontrol agent Colletotrichum gloeosporioides. * METHODS: To determine the changes in cell wall composition, we used semi-thin sections and a battery of antibody probes that recognize components of the cell wall and immunogold-silver cytochemistry to visualize the probes. * KEY RESULTS: A loss of specific plant cell wall polysaccharides in the region surrounding the primary fungal infection and the creation of a defensive layer by the plant to limit the fungal invasion were the two most obvious changes noted in this study. At the invasion site, there was significant loss of rhamnogalacturon-1 (RGI) and esterified and de-esterified homogalacturonan (HG)-reactive epitopes from the cell walls. In contrast, boundary tissue between the vascular tissue and the fungal lesion reacted more strongly with antibodies that recognize arabinogalactan proteins (AGPs) and xyloglucans than in unaffected areas. * CONCLUSIONS: These data strongly indicate a role of pectinases in the invasion of the biocontrol agent and the importance of extensins, AGPs, and xyloglucans as defense by the host. PMID- 21616841 TI - Intrinsic and extrinsic hydraulic factors in varying sizes of two Amazonian palm species (Iriartea deltoidea and Mauritia flexuosa) differing in development and growing environment. AB - PREMISE OF THE STUDY: This study seeks to determine how hydraulic factors vary with ontogeny and whether they begin to limit further height growth in palms. Palms are an attractive group for physiological research because their columnar trunks and simple leaf habit allow key intrinsic and extrinsic hydraulic variables to be estimated more easily than in complex arborescent dicotyledons. * METHODS: We measured various biometric and physiological factors including sap flux, leaf areas, turnover rates, and internode lengths in two Amazonian rainforest species: terra firme Iriartea deltoidea and swamp-adapted Mauritia flexuosa. These two palm species differ markedly in edaphic conditions, leaf type (pinnately compound vs. palmate), and bole development, making physiological comparisons between them important as well. * KEY RESULTS: The species exhibited differing patterns in height growth rate along boles, which appear to relate to their differences in bole development. Growth rates ultimately slowed at the tops of tall palms in both species. We also found a high degree of convergence in total leaf area with height in both species even though they exhibited contrasting patterns in both live frond number and leaf area per frond with height. Sap flux density from leaves was constant with height but four times greater in M. flexuosa than in I. deltoidea. * CONCLUSIONS: Although height growth rates slow considerably in tall palms, neither species shows evidence that hydraulic factors become limiting because they are able to support much greater leaf areas with similar sap flux densities as shorter palms. PMID- 21616842 TI - The importance of soil microorganisms for maintaining diverse plant communities in tallgrass prairie. AB - PREMISE OF THE STUDY: According to the "Janzen-Connell hypothesis," soil microorganisms have the potential to increase plant community diversity by mediating negative feedback on plant growth. Evidence for such microbe-driven negative feedback has been found in a variety of terrestrial systems. However, it is currently unknown how general this phenomenon is within most plant communities. Also unknown is the role of mutualists in generating such feedback: do they decrease the influence of soil-mediated negative feedback on plant fitness or do they increase its effect by proliferating with plant hosts to which they give the least benefit? * METHODS: We investigated soil-microbe-mediated feedback via a series of reciprocal transplant experiments in the greenhouse using soil from a restored tallgrass prairie and native tallgrass prairie plant species. * KEY RESULTS: We found that negative feedback was very common but that mutualists (arbuscular mycorrhizal fungi) influence plant growth in opposition to the overall negative feedback trend. * CONCLUSIONS: Widespread microbially mediated negative feedback indicates that plant community diversity and composition in tallgrass prairie are dependent on soil microorganisms. Native soil microorganisms should be considered in restoration efforts of tallgrass prairie and, potentially, other native plant communities. PMID- 21616843 TI - Why does Viola hondoensis (Violaceae) shed its winter leaves in spring? AB - PREMISE OF THE STUDY: Viola hondoensis is a perennial herb that inhabits the understory of temperate, deciduous forests. It is an evergreen plant with a leaf life span that is shorter than a year. Its summer leaves are produced in spring and shed in autumn; winter leaves are produced in autumn and shed in spring. Here we asked why the plant sheds its winter leaves in spring, though climate conditions improve from spring to summer. We proposed four hypotheses for the cause of shedding: (1) changes in seasonal environment such as day length or air temperature, (2) shading by canopy deciduous trees, (3) self-shading by taller summer leaves, and (4) competition for nutrients between summer and winter leaves. * METHODS: To test these hypotheses, we manipulated the environment of winter leaves: (1) plants were transplanted to the open site where there was no shading by canopy trees. (2) Petioles of summer leaves were anchored to the soil surface to avoid shading of winter leaves. (3) Sink organs were removed to eliminate nutrient competition. * KEY RESULTS: Longevity of winter leaves was extended when shading by summer leaves was eliminated and when sink organs were removed, but not when plants were transplanted to the open site. * CONCLUSION: We conclude that the relative difference in light availability between summer and winter leaves is a critical factor for regulation of leaf shedding, consistent with the theory of maximization of the whole-plant photosynthesis. PMID- 21616844 TI - Root contraction helps protect the "living rock" cactus Ariocarpus fissuratus from lethal high temperatures when growing in rocky soil. AB - PREMISE OF THE STUDY: We investigated how the "living rock" cactus Ariocarpus fissuratus, like other low-growing desert plants, can endure potentially lethal high temperatures at the soil surface. Specifically, we examined how shoot descent by root contraction in the presence or absence of soil rocks influences shoot temperatures and transpiration. * METHODS: Root contraction was identified by measuring shoot descent and anatomical analysis. Temperatures and transpiration were measured for plants at two heights in sandy and rocky soil, and temperature tolerances were determined by vital staining. * KEY RESULTS: Plants embedded in rocky soil survived an extreme heat episode, unlike plants in sandy soil, though rocks did not moderate low temperatures. Root contraction occurred regardless of season and soil moisture. Xylem conduits (wide-band tracheids) formed a compressible lattice that decreased root length as rays enlarged the root base radially. Plant position in the soil did not affect transpiration. * CONCLUSIONS: Contractile roots pulled plants of A. fissuratus into the soil at rates of 6-30 mm yr(-1). Maintaining shoots level with the soil surface kept plant temperatures below the high lethal temperature and improved survivorship in soil shaded by surface rocks. PMID- 21616845 TI - Molecular systematics and morphological character evolution of the Condamineeae (Rubiaceae). AB - PREMISE OF THE STUDY: The Condamineeae have in previous molecular studies been shown to be part of an early-divergent clade within the subfamily Ixoroideae, together with the tribes Calycophylleae, and Hippotideae, and genera of the former Cinchoneae and Rondeletieae. Generic relationships within this clade have, however, remained largely unresolved. * METHODS: In this study, the systematics of the Condamineeae was further examined by phylogenetic reconstruction of six cpDNA regions and one nrDNA region using parsimony and Bayesian Markov chain Monte Carlo inference. Morphological character evolution within the tribe was assessed by ancestral state reconstruction using likelihood optimization of characters onto Bayesian trees. * KEY RESULTS: Calycophylleae appears polyphyletic. "Hippotideae" is monophyletic but nested within the Condamineeae. The phylogenetic hypotheses presented support a resurrection of the genera Holtonia, Schizocalyx, and Semaphyllanthe. Furthermore, Bathysa is found to be polyphyletic, Tresanthera is found nested within Rustia, and the taxonomically disputed genus Dialypetalanthus is here shown to be sister to a Bothriospora Wittmackanthus clade. Morphological ancestral state reconstructions indicate that protogyny have evolved at least two times within the tribe and that indehiscent fruits, loculicidal fruit dehiscence, and intrapetiolar stipules have evolved independently several times. The occurrence of calycophylls (leaf-like calyx lobes), poricidal anthers, and winged seeds also appear homoplastic within the tribe. * CONCLUSIONS: A diagnosis and delimitation of the tribe Condamineeae is presented, with taxonomic proposals to synonymize Tresanthera and to transfer several species of Bathysa as well as Phitopis to a resurrected Schizocalyx. PMID- 21616846 TI - Fruits and foliage of Pueraria (Leguminosae, Papilionoideae) from the Neogene of Eurasia and their biogeographic implications. AB - PREMISE OF THE STUDY: Pueraria (Leguminosae, Papilionoideae) is native in East Asia, South Asia, Southeast Asia, and Oceania and is well known as a rampant invasive weed in the southeastern United States (P. montana; better known as kudzu), but relatively little is known about its early evolution and biogeographic origin. * METHODS: On the basis of comparative analyses of the fruit and leaflet architecture of closely related extant and fossil taxa, we studied the fossil history and biogeography of Pueraria. * KEY RESULTS: Fossil Pueraria is recognized on the basis of distinctive fruit and foliage from the Mio Pliocene of middle latitudes in China, Japan, Abkhazia, and Croatia. Recognition of P. miothunbergiana from the Mio-Pliocene of China and Japan is reinforced by a trifoliolate leaf as well as isolated lateral and terminal leaflets. Pueraria shanwangensis sp. nov. represents the first recognition of fossil Pueraria fruits. This fruit species co-occurs with P. miothunbergiana in the Middle Miocene Shanwang flora and possibly represents the same population. Pueraria maxima (Unger) comb. nov., previously named as Dolichites maximus or Desmodium maximum, is recognized on the basis of leaflets from the Miocene of Croatia and Abkhazia. Other prior fossil reports of Pueraria and Dolichites are reevaluated. * CONCLUSIONS: Pueraria had begun to diversify by at least the Middle Miocene and had spread into the Mio-Pliocene subtropical and temperate floras of the Balkan Peninsula, the Caucasus, and eastern Asia, which suggests the present diversity of this genus in tropical Asia and Oceania might have originated from the mid latitudes of Eurasia. PMID- 21616847 TI - Pollen-mediated gene flow in isolated and continuous stands of bur oak, Quercus macrocarpa (Fagaceae). AB - PREMISE OF THE STUDY: Pollination patterns determine the reproductive neighborhood size of plants, the connectivity of populations, and the impacts of habitat fragmentation. We characterized pollination in three populations of Quercus macrocarpa occurring in a highly altered landscape in northeastern Illinois to determine whether isolated remnant stands were reproductively isolated. * METHODS: We used microsatellites to genotype all adults and 787 acorns from two isolated savanna remnants and a stand in an old-growth forest. One isolated remnant occurred in a highly urbanized/industrialized landscape, and one occurred in an agricultural landscape. Parentage assignment was used to assess pollen-mediated gene flow. * KEY RESULTS: Pollen donors from outside the study sites accounted for between 46% and 53% of paternities and did not differ significantly among sites, indicating that similar high levels of gene flow occurred at all three sites. Within stands, the mean pollination distance ranged from 42 to 70 meters, and when accounting for outside pollinations, mean pollination distances were well over 100 meters. Genetic diversity of incoming pollen was extremely high in all three stands. The number of effective pollen donors, N(ep), calculated from paternity assignment was higher than that estimated by an indirect correlated paternity approach. * CONCLUSIONS: Our findings indicate that extremely isolated stands of oaks are unlikely to be genetically and reproductively isolated, and remnant stands may contribute to maintaining genetic connectivity in highly modified landscapes. PMID- 21616848 TI - Hydraulic properties of fern sporophytes: Consequences for ecological and evolutionary diversification. AB - PREMISE OF THE STUDY: Ferns are an important component of both tropical and temperate forests; yet, our understanding of the water relations of their sporophyte generation is limited. Indeed, to date there has been no large scale survey that attempts to clarify how ferns fit into current ideas of plant water relations. This study examines several tropical ferns with the goal of understanding how these characters vary between species from various habitats and across life forms * METHODS: We measured stipe hydraulic conductivity, water potential, and vulnerability to cavitation along with photosynthetic variables and leaf allometry of 21 species from 14 genera to identify physiological trait assemblages across taxa. * KEY RESULTS: Epiphytic ferns have significantly lower hydraulic conductivity and a vascular system more resistant to cavitation (i.e., higher P(50) values). They reached lower mid-day water potentials and produced leaves with reduced stipe lengths and reduced laminar area relative to terrestrial species. Xylem specific hydraulic conductivity (K(S)) was correlated with the mean hydraulic diameter of tracheids in terrestrial species, but not in epiphytes. There was no evidence of safety-efficiency trade-offs in any group. * CONCLUSIONS: When compared across life forms, our data shed light on physiological mechanisms that may have allowed for terrestrial ferns to move into the epiphytic habit. When compared across a diverse assemblage of terrestrial plants, we find that resistance to water flow in fern stipes is significantly higher than that recorded from the stems of seed plants. PMID- 21616849 TI - Geographic variation in reproductive success of Stenocereus thurberi (Cactaceae): Effects of pollination timing and pollinator guild. AB - PREMISE OF THE STUDY: It has been proposed that species of columnar cacti from dry tropical areas depend on bats for their reproduction, whereas species from dry subtropical areas are also pollinated by other species. To test this hypothesis, we examined the effects of pollinator guild and of variation in time and space on the reproductive success of a widespread species. * METHODS: Changes in fruit set, seed set, and pollinator activity through time were recorded in three widely separated populations of Stenocereus thurberi. Breeding system and sources of pollination limitation were determined by controlled pollinator exclusions in each population. * KEY RESULTS: Significant differences were found in the timing of activity and in the effectiveness of pollinators among sites. In the northern and central populations, reproductive success depends on bats, whereas in the southern population a combination of pollinators was more effective. No difference between open and hand cross-pollination treatments was found in the northern and central populations, which suggests no pollen limitation. However, significant differences were detected in the southern population, which indicates temporal differences in pollinator abundance or arrival time. * CONCLUSIONS: Local variation in pollinator assemblages and reproductive success could greatly affect the evolution of pollination systems. The pattern of generalist pollination in the southernmost populations and specialized pollination in the central and northern populations contradicts the hypothesis of latitudinal variation. In the absence of nocturnal pollinators, the accumulated nectar can sustain visits by diurnal pollinators, a bet-hedging strategy that increases the chances of fruit set in some populations. PMID- 21616850 TI - A complete generic phylogeny of Malpighiaceae inferred from nucleotide sequence data and morphology. AB - PREMISE OF THE STUDY: The Malpighiaceae include ~1300 tropical flowering plant species in which generic definitions and intergeneric relationships have long been problematic. The goals of our study were to resolve relationships among the 11 generic segregates from the New World genus Mascagnia, test the monophyly of the largest remaining Malpighiaceae genera, and clarify the placement of Old World Malpighiaceae. * METHODS: We combined DNA sequence data for four genes (plastid ndhF, matK, and rbcL and nuclear PHYC) from 338 ingroup accessions that represented all 77 currently recognized genera with morphological data from 144 ingroup species to produce a complete generic phylogeny of the family. * KEY RESULTS AND CONCLUSIONS: The genera are distributed among 14 mostly well supported clades. The interrelationships of these major subclades have strong support, except for the clade comprising the wing-fruited genera (i.e., the malpighioid+Amorimia, Ectopopterys, hiraeoid, stigmaphylloid, and tetrapteroid clades). These results resolve numerous systematic problems, while others have emerged and constitute opportunities for future study. Malpighiaceae migrated from the New to Old World nine times, with two of those migrants being very recent arrivals from the New World. The seven other Old World clades dispersed much earlier, likely during the Tertiary. Comparison of floral morphology in Old World Malpighiaceae with their closest New World relatives suggests that morphological stasis in the New World likely results from selection by neotropical oil-bee pollinators and that the morphological diversity found in Old World flowers has evolved following their release from selection by those bees. PMID- 21616851 TI - Ecogeography of ploidy variation in cultivated potato (Solanum sect. Petota). AB - PREMISE OF THE STUDY: The taxonomy of cultivated potatoes has been highly controversial, with estimates of species numbers ranging from 3 to 17. Ploidy level has been one of the most important taxonomic characters to recognize cultivated potato species, containing diploid (2n = 2x = 24), triploid (2n = 3x = 36), tetraploid (2n = 4x = 48), and pentaploid (2n = 5x = 60) cultivars. We tested the environmental associations of different ploidy levels in cultivated potato species that traditionally have been recognized as Linnaean taxa to see whether, in combination with prior morphological, molecular, and crossing data, some of the ploidy variants can be recognized as distinct taxa. * METHODS: We summarize 2780 chromosome counts of landrace cultivated potatoes, provide georeferences to 2048 of them, and analyze these data for 20 environmental variables at 10-min resolution using the randomForest algorithm to explore associations with taxa and ploidy variants. * KEY RESULTS: Except for the S. tuberosum Chilotanum Group and extreme northern and southern range extensions of the Andigenum Group, it is impossible to find distinct habitats for the ploidy variants of the S. tuberosum Andigenum Group. * CONCLUSIONS: Our distributional and ecological data, in combination with prior results from morphology, microsatellites, and crossing data, provide yet additional data to support a major reclassification of cultivated potato species. A rational, stable, and universally accepted taxonomy of this major crop plant will greatly aid all users of wild and cultivated potatoes from breeders to gene bank managers to ecologists and evolutionary biologists. PMID- 21616852 TI - Species-specific SSR alleles for studies of hybrid cattails (Typha latifolia x T. angustifolia; Typhaceae) in North America. AB - PREMISE: Studies of hybridizing species are facilitated by the availability of species-specific molecular markers for identifying early- and later-generation hybrids. Cattails are a dominant feature of wetland communities, and a better understanding of the prevalence of hybrids is needed to assess the ecological and evolutionary effects of hybridization. Hybridization between Typha angustifolia and T. latifolia produce long-lived clones, known as Typha *glauca, which are considered to be invasive. Although morphological variation in cattails makes it difficult to recognize early- and later-generation hybrids, several dominant, species-specific RAPD markers are available. Our goal was to find codominant, species-specific markers with greater polymorphism than RAPDs, to identify later generation hybrids more efficiently. * METHODS: We screened nine SSR (simple sequence repeat) loci that were described from populations in Ukraine, and we surveyed 31 cattail populations from the upper Midwest and eastern USA. * KEY RESULTS: Seven SSR loci distinguished the parent taxa and were consistent with known species-specific RAPD markers, allowing easier detection of backcrossing. We used linear discriminant analysis to show that F(1) hybrid phenotypes were intermediate between the parent taxa, while those of backcrossed plants overlapped with the hybrids and their parents. Log(leaf length/leaf width), spike gap length, spike length, and stem diameter explained much of the variation among groups. * CONCLUSIONS: We provide the first documentation of backcrossed plants in hybridizing cattail populations in Michigan. The diagnostic SSR loci we identified should be extremely useful for examining the evolutionary and ecology interactions of hybridizing cattails in North America. PMID- 21616853 TI - New microsatellite loci for pomegranate, Punica granatum (Lythraceae). AB - PREMISE OF THE STUDY: A new set of pomegranate microsatellites was selected and characterized to assess the level of genetic diversity among cultivars and wild genotypes. * METHODS AND RESULTS: Nine Simple Sequence Repeat (SSR) markers were obtained using the Microsatellite-AFLP technique and were successfully amplified in 34 genotypes belonging to Italian, Spanish, and Turkish germplasm collections. The number of alleles per locus ranged from 1 to 5, and the total number of alleles was 22. * CONCLUSIONS: Because only a few codominant markers are available for this species, the newly identified SSRs will facilitate genetic diversity studies, fingerprinting, and mapping. In addition, the 9 loci successfully amplified in P. granatum var. nana. No cross transferability was observed for Cuphea micropetala and Lagerstroemia indica (Lythraceae). PMID- 21616854 TI - Ten microsatellite markers in endangered species Sauvagesia rhodoleuca (Ochnaceae). AB - PREMISE OF THE STUDY: We developed microsatellite markers to investigate the level of genetic diversity within and among populations of the endemic shrub Sauvagesia rhodoleuca in China. * METHODS AND RESULTS: Ten polymorphic microsatellite loci were identified in five populations. The number of alleles per locus varied from 2 to 16. Observed and expected heterozygosities ranged from 0.000 to 1.000 and from 0.000 to 0.726, respectively. * CONCLUSIONS: The results provide basic information on genetic diversity for future studies of population genetics in S. rhodoleuca. PMID- 21616855 TI - A set of microsatellite markers for Arrabidaea chica (Bignoniaceae), a medicinal liana from the Neotropics. AB - PREMISE OF THE STUDY: Microsatellite markers were developed, optimized, and characterized for Arrabidaea chica (Humb. & Bonpl.) Verl. (Bignoniaceae), a Neotropical liana extensively used in folk medicine. The aim of this study was to develop molecular tools to investigate the genetic structure and diversity of natural populations and germplasm collections of this species. * METHODS AND RESULTS: Eight highly polymorphic microsatellite markers revealed a multibanded pattern, suggesting that the species is polyploid. The total number of bands per locus ranged from 9 to 17, revealing high levels of polymorphism. * CONCLUSIONS: The high level of polymorphism detected with these markers indicates their utility in devising conservation strategies and rational exploitation of A. chica. PMID- 21616856 TI - A rapid and inexpensive method for the direct PCR amplification of DNA from plants. AB - PREMISE OF THE STUDY: We present a rapid and inexpensive alternative to DNA isolation for polymerase chain reaction (PCR) amplification from plants. * METHODS AND RESULTS: The method involves direct PCR amplification from material macerated in one buffer, followed by dilution and incubation in a second buffer. We describe the procedure and demonstrate its application for nuclear and plastid DNA amplification across a broad range of vascular plants. * CONCLUSIONS: The method is fast, easy to perform, cost-effective, and consequently ideal for large sample numbers. It represents a considerable simplification of present approaches requiring DNA isolation prior to PCR amplification and will be useful in plant systematics and biotechnology, including applications such as DNA barcoding. PMID- 21616857 TI - Generation and characterization of 11 novel est derived microsatellites from Vicia faba (Fabaceae). AB - PREMISE OF THE STUDY: Simple sequence repeat (SSR) markers were developed for faba bean using expressed sequence tags (ESTs) from the NCBI database to study for genetic diversity. * METHODS AND RESULTS: A total of 11 novel EST-SSR loci were generated and characterized when tested on four populations of 29 faba bean individuals from China and Europe. The number of alleles (A) ranged from 1 to 3 in each population, and observed heterozygosity (H(O)) and expected heterozygosity (H(E)) ranged from 0 to 0.5000 and 0.6400, respectively. Furthermore, transferable analysis revealed that eight of these loci (72.73%) amplified in Pisum sativum L., six of which (75.00%) detected polymorphism. * CONCLUSIONS: The developed markers in this study will provide valuable tools for genetic diversity, resource conservation, genetic mapping, and marker-assisted breeding of faba bean in the future. PMID- 21616858 TI - "Prototaxites was not a taphonomic artifact". PMID- 21616859 TI - The enigmatic Devonian fossil Prototaxites is not a rolled-up liverwort mat: Comment on the paper by Graham et al. (AJB 97: 268-275). PMID- 21616860 TI - Rolled liverwort mats explain major Prototaxites features: Response to commentaries. AB - In volume 97(2) of the American Journal of Botany (pp. 268-275), we published an article entitled "Structural, physiological, and stable carbon isotopic evidence that the enigmatic Paleozoic fossil Prototaxites formed from rolled liverwort mats". Here, we respond to a letter and a commentary on our article in the present issue, welcoming this opportunity to continue the scientific dialogue about an issue that has long been stimulating and controversial. For the reader's benefit, we first briefly describe the recent scholarly context of our article. PMID- 21616861 TI - The hydrostatic gradient, not light availability, drives height-related variation in Sequoia sempervirens (Cupressaceae) leaf anatomy. AB - PREMISE OF THE STUDY: Leaves at the tops of most trees are smaller, thicker, and in many other ways different from leaves on the lowermost branches. This height related variation in leaf structure has been explained as acclimation to differing light environments and, alternatively, as a consequence of hydrostatic, gravitational constraints on turgor pressure that reduce leaf expansion. * METHODS: To separate hydrostatic effects from those of light availability, we used anatomical analysis of height-paired samples from the inner and outer tree crowns of tall redwoods (Sequoia sempervirens). * KEY RESULTS: Height above the ground correlates much more strongly with leaf anatomy than does light availability. Leaf length, width, and mesophyll porosity all decrease linearly with height and help explain increases in leaf-mass-to-area ratio and decreases in both photosynthetic capacity and internal gas-phase conductance with increasing height. Two functional traits-leaf thickness and transfusion tissue also increase with height and may improve water-stress tolerance. Transfusion tissue area increases enough that whole-leaf vascular volume does not change significantly with height in most trees. Transfusion tracheids become deformed with height, suggesting they may collapse under water stress and act as a hydraulic buffer that improves leaf water status and reduces the likelihood of xylem dysfunction. * CONCLUSIONS: That such variation in leaf structure may be caused more by gravity than by light calls into question use of the terms "sun" and "shade" to describe leaves at the tops and bottoms of tall tree crowns. PMID- 21616862 TI - Heterogeneous selection on trichome production in Alaskan Arabidopsis kamchatica (Brassicaceae). AB - PREMISE OF THE STUDY: Environmental heterogeneity is thought to be one of the primary factors in the evolutionary maintenance of morphological variation. Here, we explore the role of environmental heterogeneity in the maintenance of variation in leaf hair (trichome) production in Arabidopsis kamchatica. * METHODS: We investigate abiotic correlates of trichome production in A. kamchatica via surveys of both herbarium specimens and wild populations. In addition, we examine patterns of phenotypic selection on trichome production among populations that differ in environmental characteristics. * KEY RESULTS: Trichome-producing herbarium specimens were more likely to occur at lower latitudes and in locations with lower mean annual precipitation and less annual variation in temperature than glabrous specimens. In surveys of wild populations, frequencies of trichome-producing plants were higher in drier habitats than in wetter environments. Using phenotypic selection analysis, we found divergent selection through female fitness (fruit production) on trichome number in populations that differ in environmental characteristics; there was selection for reduced trichome number in one population and selection for increased trichome number in another population. In a population containing both glabrous and trichome-producing plants, glabrous plants produced significantly more fruits than trichome-producing individuals, which indicates selection against the trichome morph. * CONCLUSIONS: Our results demonstrate that there is heterogeneity in selection among populations, which could be responsible for the maintenance of trichome variation in Alaskan populations of A. kamchatica. PMID- 21616863 TI - Variation in leaf stomatal traits of 28 tree species in relation to gas exchange along an edaphic gradient in a Bornean rain forest. AB - PREMISE OF THE STUDY: Quantifying variation in functional traits associated with shifts in the species composition of plant communities along resource gradients helps identify environmental attributes important for community assembly. Stomates regulate the balance between carbon assimilation and water status in plants. If environmental attributes affecting photosynthetic water-use efficiency govern species distribution along an edaphic gradient, then adaptive variation in stomatal traits of plant species specializing on different soils should reflect belowground resource availability. * METHODS: We tested this hypothesis by quantifying stomatal trait variation in understory saplings of 28 Bornean tree species in relation to gas exchange and water-use efficiency (WUE). * KEY RESULTS: Comparisons between congeneric specialists of the more fertile, moister clay and the less fertile, well-drained sandy loam revealed little evidence of similar shifts in stomatal traits across genera, nor was stomatal pore index correlated with g(max), A(max), or WUE (A(max)/g(max) or Delta(13)C), suggesting that stomates may be overbuilt in these shaded juveniles. A(max) was higher on sandy loam, likely due to higher understory irradiance there, but there were no other significant differences in gas exchange or WUE. * CONCLUSIONS: Despite substantial diversity in stomatal anatomy, there were few strong relationships between stomatal, photosynthetic, and WUE traits in relation to soil resources. Routine differences in water availability therefore may not exert a dominant control on the distributions of these Bornean tree species. Furthermore, the clades represented by these 12 genera may possess alternative functional designs enabling photosynthetic WUE that is sufficient to these humid, understory environments, due to whole plant-functional integration of stomatal traits with other, unmeasured traits influencing gas exchange. PMID- 21616864 TI - Polystichum munitum (Dryopteridaceae) varies geographically in its capacity to absorb fog water by foliar uptake within the redwood forest ecosystem. AB - PREMISE OF THE STUDY: Fog provides a critical water resource to plants around the world. In the redwood forest ecosystem of northern California, plants depend on fog absorbed through foliar uptake to stay hydrated during the rainless summer. In this study, we identified regions within the redwood ecosystem where the fern Polystichum munitum canopy most effectively absorbs fog drip that reaches the forest floor. * METHODS: We measured the foliar uptake capacity of P. munitum fronds at seven sites along 700 km of the redwood forest ecosystem. We quantified the canopy cover of P. munitum at each site and estimated how much water the fern canopy can acquire aboveground through fog interception and absorption. * KEY RESULTS: Throughout the ecosystem, nocturnal foliar uptake increased the leaf water content of P. munitum by 7.2%, and we estimated that the P. munitum canopy can absorb 5 +/- 3% (mean +/- SE) of intercepted fog precipitation. Strikingly, P. munitum had the highest foliar uptake capacity in the center of the ecosystem and may absorb 10% more of the fog its canopy intercepts in this region relative to other regions studied. Conversely, P. munitum had no foliar uptake capacity in the southern end of the ecosystem. * CONCLUSIONS: This study shows the first evidence that foliar uptake varies within species at the landscape scale. Our findings suggest that the P. munitum at the southern tip of the redwood ecosystem may suffer most from low summertime water availability because it had no potential to acquire fog as an aboveground water subsidy. PMID- 21616865 TI - The evolution of Cayaponia (Cucurbitaceae): Repeated shifts from bat to bee pollination and long-distance dispersal to Africa 2-5 million years ago. AB - PREMISE OF THE STUDY: The Cucurbitaceae genus Cayaponia comprises ~60 species that occur from Uruguay to the southern United States and the Caribbean; C. africana occurs in West Africa and on Madagascar. Pollination is by bees or bats, raising the question of the evolutionary direction and frequency of pollinator shifts. Studies that investigated such shifts in other clades have suggested that bat pollination might be an evolutionary end point. * METHODS: Plastid and nuclear DNA sequences were obtained for 50 accessions representing 30 species of Cayaponia and close relatives, and analyses were carried out to test monophyly, infer divergence times, and reconstruct ancestral states for habitat preferences and pollination modes. * KEY RESULTS: The phylogeny shows that Cayaponia is monophyletic as long as Selysia (a genus with four species from Central and South America) is included. The required nomenclatural transfers are made in this paper. African and Madagascan accessions of C. africana form a clade that is part of a polytomy with Caribbean and South American species, and the inferred divergence time of 2-5 Ma implies a transoceanic dispersal event from the New World to Africa. The ancestral state reconstructions suggest that Cayaponia originated in tropical forests from where open savannas were reached several times and that bee pollination arose from bat pollination, roughly concomitant with the shifts from forests to savanna habitats. * CONCLUSIONS: Cayaponia provides the first example of evolutionary transitions from bat to bee pollination as well as another instance of transoceanic dispersal from the New World to Africa. PMID- 21616866 TI - Phylogeny and character evolution in Medicago (Leguminosae): Evidence from analyses of plastid trnK/matK and nuclear GA3ox1 sequences. AB - PREMISE OF THE STUDY: The genus Medicago, with about 87 species, includes the model legume species M. truncatula, and a number of important forage species such as M. sativa (alfalfa), M. scutellata (snail medic), and M. lupulina (black medic). Relationships within the genus are not yet sufficiently resolved, contributing to difficulty in understanding the evolution of a number of distinguishing characteristics such as aneuploidy and polyploidy, life history, structure of cotyledons, and number of seeds per fruit. * METHODS: Phylogenetic relationships of 70-73 species of Medicago and its sister genus Trigonella (including Melilotus) were reconstructed from nucleotide sequences of the plastid trnK/matK region and the nuclear-encoded GA3ox1 gene (gibberellin 3-beta hydroxylase) using maximum parsimony and Bayesian inference methods. * KEY RESULTS: Our results support certain currently recognized taxonomic groups, e.g., sect. Medicago (with M. sativa) and sect. Buceras. However, other strongly supported clades-the "reduced subsection Leptospireae clade" that includes M. lupulina, the "polymorpha clade" that includes M. murex and M. polymorpha and the "subsection Pachyspireae clade" that includes M. truncatula-each of which includes species presently in different subsections of sect. Spirocarpos, contradict the current classification. * CONCLUSIONS: These results support the hypothesis that some characters considered important in existing taxonomies, for example, single-seeded fruits that have arisen more than once in both Medicago and Trigonella, are indeed homoplastic. Others, such as the 2n = 14 chromosome number, have also arisen independently within the genus. In addition, we demonstrate support for the utility of GA3ox1 sequences for phylogenetic analysis among and within closely related genera of legumes. PMID- 21616867 TI - Hybridization and polyploidy of an aquatic plant, Ruppia (Ruppiaceae), inferred from plastid and nuclear DNA phylogenies. AB - PREMISE OF THE STUDY: The monogeneric family Ruppiaceae is found primarily in brackish water and is widely distributed on all continents, many islands, and from subartic to tropical zones. Ruppia taxonomy has been confusing because of its simplified morphology yet high phenotypic plasticity and the existence of polyploidy and putative hybrids. This study addresses the current classification of species in the genus, the origin of putative hybrids and polyploids, and the distribution of Ruppia species. * METHODS: Separate molecular phylogenetic analyses using plastid DNA and nuclear-encoded PHYB data sets were performed after chromosome observations. * KEY RESULTS: The resultant trees were largely congruent between genomes, but were incongruent in two respects: the first incongruence may be caused by long outgroup branches and their effect on ingroup rooting, and the second is caused by the existence of heterogeneous PHYB sequences for several accessions that may reflect several independent hybridization events. Several morphological species recognized in previous taxonomic revisions appear paraphyletic in plastid DNA and PHYB trees. * CONCLUSIONS: Given the molecular phylogenies, and considering chromosome number and morphology, three species and one species complex comprising six lineages were discerned. A putative allotriploid, an allotetraploid, and a lineage of hybrid origin were identified within the species complex, and a hybrid was found outside the species complex, and their respective putative parental taxa were inferred. With respect to biogeography, a remarkably discontinuous distribution was identified in two cases, for which bird-mediated seed dispersal may be a reasonable explanation. PMID- 21616868 TI - Molecular phylogeny of the genus Vitis (Vitaceae) based on plastid markers. AB - PREMISE OF THE STUDY: This work represents the first molecular phylogeny of the economically important genus Vitis, an important genetic resource for breeding in grapevine, Vitis vinifera. * METHODS: A molecular phylogeny of Vitis using a combined data set of three noncoding regions of the plastid DNA genome was constructed from 47 accessions covering 30 species of Vitis. The data for the trnL-F marker were combined with previously published data across the Vitaceae. * KEY RESULTS: The molecular phylogeny demonstrated monophyly of the genus Vitis. Based on the combined analysis of three genes, Vitis is split into three clades that mirror the continental distribution of these accessions. The diversity is highest in the Asian clade, but the general genetic distances across taxa from different continents are relatively small. * CONCLUSIONS: The findings support a relatively recent and intense gene flow between East Asia and North America and the possible impact of hybridization on the evolution of the genus Vitis. Taxon identity in important stock collections should be screened carefully because roughly 10% of the accessions analyzed in the present study had been misidentified. PMID- 21616869 TI - Upper Cretaceous woods from the Olmos Formation (late Campanian-early Maastrichtian), Coahuila, Mexico. AB - PREMISE OF THE STUDY: The Olmos Formation was part of a system of deltas that existed in the southern portion of the Western Interior of North America during the Campanian-Maastrichtian. The paleofloristic composition from the northern portions of the Epicontinental Sea is relatively well known, but less intensive exploration in the south has precluded more detailed floristic comparison across the entire latitudinal span of the Sea. The Olmos Formation flora, with more than 100 different leaf morphotypes so far recognized and several wood types, has the most diverse Cretaceous fossil plant assemblage in Mexico and represents a valuable opportunity for comparative studies. * METHODS: The fossil woods here described were collected in the Coahuila State, Mexico. The samples were studied using standard thin section technique and identified by comparison with fossil and extant material. * KEY RESULTS: We described four new genera (Olmosoxylon, cf. Lauraceae; Coahuiloxylon, ?Anacardiaceae, ?Burseraceae; Muzquizoxylon, Cornaceae; and Wheeleroxylon, Malvaceae s.l.) and three xylotypes of angiosperms. * CONCLUSIONS: Some of the genera present in the Olmos Formation such as Javelinoxylon and Metcalfeoxylon have been described from geologic units in the USA (San Juan Basin, New Mexico and Big Bend National Park, Texas), suggesting similarity in the taxonomic composition of the floras that inhabited southern portions of the western margin of the Campanian-Maastrichtian Epicontinental Sea. Other species, however, have only been reported for the Olmos Formation, indicating some degree of local floristic differentiation among the assemblages that inhabited the southern portion of the Western Interior. PMID- 21616870 TI - Polyphenol oxidase activity in the roots of seedlings of Bromus (Poaceae) and other grass genera. AB - PREMISE OF THE STUDY: Phenolic compounds exuded by roots have been implicated in allelopathic interactions among plants. Root enzymes that destroy phenolics may protect plants against allelopathic inhibition and thus may aid in invasiveness. Phenolic-degrading enzymes are chiefly found in aboveground plant parts, but have also been previously reported in root tissues where the enzyme's function is unknown. We explored phenolic oxidase activity in emerging roots of grasses in a survey across different grass genera; in particular, we aimed to test whether grasses of the genus Bromus, known for their large invasion potential, differ in this respect from other grass taxa. * METHODS: We assayed a range of grass genera commonly found in the United States for root enzyme activity with spectrophotometric assays of phenol oxidase activity using l-DOPA as the main substrate. * KEY RESULTS: In the survey of a grass genera, we discovered that roots of the genus Bromus contain large amounts of polyphenol oxidase (PPO) activity, while all other tested grass genera, even ones closely related to Bromus, did not. PPO was found to be present at germination and remained active throughout the life of the plant. Compared to other PPOs, the enzyme present in Bromus appears to have a narrow substrate range. * CONCLUSIONS: The specific functions of the root PPO and the ecological ramifications of the special status of Bromus are not yet clear. The possibility that the enzyme plays a role in plant species interaction for bromes, a genus of grasses known to have high invasive potential, is raised. PMID- 21616871 TI - Spatial genetic structure within size classes of the endangered tropical tree Guaiacum sanctum (Zygophyllaceae). AB - PREMISE OF THE STUDY: Patterns of spatial genetic structure (SGS) were analyzed within a population of the endangered tropical tree Guaiacum sanctum located in northwestern Costa Rica. Documentation of these patterns provides insights into the gene dispersal mechanisms that play a central role in the maintenance and structure of genetic diversity within plant populations. * METHODS: Allozyme analyses were used to examine SGS in Palo Verde National Park, Costa Rica. The SGS was compared among three plots and different age classes. * KEY RESULTS: High levels of genetic diversity were found overall with a pooled genetic diversity of H(e) = 0.302 (+/-0.02). Selfing was proposed as the proximate cause for significant levels of heterozygote deficiency observed across size classes and plots. An unexpected lack of SGS (r(j) < 0.02) was observed for all size classes, suggesting the mixing of seeds from several adults. A parent-pair parentage analysis indicated that at least 48% of the smaller individuals within a plot were produced by parents located at distances of at least 150 m. * CONCLUSIONS: Populations of G. sanctum are established and maintained by bird-mediated, moderate- to long-distance seed dispersal, which results in a mixture of seeds from unrelated maternal individuals, effectively eliminating SGS. Proximity between individuals is, therefore, a poor predictor of family structure in this species. Long-distance seed dispersal, coupled with estimates of high genetic diversity, suggests that this endangered species has the potential for natural regeneration and restoration given the availability of suitable habitats. PMID- 21616872 TI - Species boundaries and frequency of hybridization in the Dryopteris carthusiana (Dryopteridaceae) complex: A taxonomic puzzle resolved using genome size data. AB - PREMISE OF THE STUDY: Genome duplication and interspecific hybridization are important evolutionary processes that significantly influence phenotypic variation, ecological behavior, and reproductive biology of land plants. These processes played a major role in the evolution of the Dryopteris carthusiana complex. This taxonomically intricate group composed of one diploid (D. expansa) and two allotetraploid (D. carthusiana and D. dilatata) species in Central Europe. Overall phenotypic similarity, great plasticity, and the incidence of interspecific hybrids have led to a continuous dispute concerning species circumscription and delimitation. * METHODS: We used flow cytometry and multivariate morphometrics to assess the level of phenotypic variation and the frequency of hybridization in a representative set covering all recognized species and hybrids. * KEY RESULTS: Flow cytometric measurements revealed unique genome sizes in all species and hybrids, allowing their easy and reliable identification for subsequent morphometric analyses. Different species often formed mixed populations, providing the opportunity for interspecific hybridization. Different frequencies of particular hybrid combinations depended primarily on evolutionary relationships, reproductive biology, and co-occurrence of progenitors. * CONCLUSIONS: Our study shows that genome size is a powerful marker for taxonomic decisions about the D. carthusiana complex and that genome size data may help to resolve taxonomic complexities in this important component of the temperate fern flora. PMID- 21616873 TI - Flower heating following anthesis and the evolution of gall midge pollination in Schisandraceae. AB - PREMISE OF THE STUDY: Flower heating is known from a few species in 11 of the c. 450 families of flowering plants. Flowers in these families produce heat metabolically and are adapted to beetles or flies as pollinators. Here, we focus on the Schisandraceae, an American/Asian plant family known to exhibit flower heating in some species, but not others, raising the question of the adaptive function of heat production. * METHODS: We used field observations, experiments, and ancestral trait reconstruction on a molecular phylogeny for Schisandraceae that includes the investigated species. * KEY RESULTS: At least two Chinese species of Illicium are exclusively pollinated by gall midges that use the flowers as brood sites (not for pollen feeding). Continuous monitoring of flower temperatures revealed that the highest temperatures were attained after the flowers' sexual functions were over, and experiments showed that post-anthetic warming benefited larval development, not fruit development. Midge larvae in flowers with trimmed tepals (and hence a lower temperature) died, but fruit set ratios remained unchanged. Based on the DNA phylogeny, gall midge pollination evolved from general fly/beetle pollination several times in Schisandraceae, with some species adapted to flower-breeding midges, others to pollen-feeding midges. * CONCLUSIONS: Flower heating may be an ancestral trait in Schisandraceae that became co-opted in species pollinated by flower-breeding midges requiring long persistent warm chambers for larval development. PMID- 21616874 TI - Natural hybridization in tropical spikerushes of Eleocharis subgenus Limnochloa (Cyperaceae): Evidence from morphology and DNA markers. AB - PREMISE OF THE STUDY: Natural hybridization represents an important force driving plant evolution and affecting community structure and functioning. Hybridization may be overlooked, however, among morphologically highly uniform congeners. An excellent example of such a group is Eleocharis subgenus Limnochloa, which has no reliably proven hybrids. Does this reflect biological barriers to interspecific crosses or difficulties in detecting the hybrids? We tested the hypothesis that hybridization occurs among sympatric Eleocharis cellulosa, E. interstincta, and E. mutata in northern Belize, Central America. * METHODS: Morphometric study (407 plants) was followed by examination of inter simple sequence repeat (ISSR) polymorphisms (44 plants) and ITS sequence variation (33 plants). * KEY RESULTS: Two putatively hybrid morphotypes were discerned-E. cellulosa-resembling and E. interstincta-resembling. DNA markers of E. cellulosa and E. interstincta displayed additive constitution in plants from one E. cellulosa-resembling population only. The other putatively hybrid populations contained ISSR and ITS markers of the species they resembled morphologically, several unique ISSR markers, and ITS sequences of an undescribed South American Limnochloa entity. DNA markers of E. mutata were absent in the putative hybrids. * CONCLUSIONS: Simultaneous use of various types of molecular markers can overcome many pitfalls of investigations concerning hybridization among closely related and morphologically similar species. Northern Belize represents a hybrid zone of E. cellulosa and E. interstincta. A third participant in the hybridization events occurring in this zone is an unknown Limnochloa lineage but is not E. mutata. Interspecific hybridization may play a significant role in the diversification of Eleocharis. PMID- 21616875 TI - Development of microsatellite markers in Lupinus luteus (Fabaceae) and cross species amplification in other lupine species. AB - PREMISE OF THE STUDY: Microsatellite primers were developed in Lupinus luteus L., an emerging temperate protein crop, to investigate genetic diversity, population structure, and to facilitate the generation of better yellow lupine varieties. * METHODS AND RESULTS: Thirteen polymorphic primer sets were evaluated in a European and Eastern European accession collection of L. luteus. The primers amplified di-, tri-, and tetranucleotide repeats with 2-4 alleles per locus. These revealed a moderate to low level of genetic variation, as indicated by an average observed heterozygosity of 0.0126. Select loci also amplified successfully in the closely related species L. hispanicus Boiss. & Reut. and in the New World species L. mutabilis Sweet. * CONCLUSIONS: These results indicate the utility of primers for the study of genetic diversity across L. luteus populations and related lupine species. The use of these microsatellite markers will facilitate the implementation of several molecular breeding strategies in yellow lupine. PMID- 21616876 TI - Development and characterization of polymorphic microsatellite markers in Momordica charantia (Cucurbitaceae). AB - PREMISE OF THE STUDY: Polymorphic microsatellite markers were developed for Momordica charantia L. to investigate the genetic diversity and population structure within and between M. charantia and its four related species (Cucurbita pepo L., Luffa cylindrical L., Lagenaria siceraria L., and Cucumis sativus L.). * METHODS AND RESULTS: Using the Fast Isolation by AFLP of Sequence COntaining Repeats (FIASCO) method, 16 polymorphic microsatellite loci were identified in 36 individuals of M. charantia. Across all the M. charantia samples, the number of alleles per locus ranged from three to eight. Seven primers successfully amplified in the four related species. * CONCLUSIONS: These markers will be useful to study population ecology and population differentiation among M. charantia species and its related species. PMID- 21616877 TI - A quantitative and statistically robust method for the determination of xylem conduit spatial distribution. AB - PREMISE OF THE STUDY: Because of their limited length, xylem conduits need to connect to each other to maintain water transport from roots to leaves. Conduit spatial distribution in a cross section plays an important role in aiding this connectivity. While indices of conduit spatial distribution already exist, they are not well defined statistically. * METHODS: We used point pattern analysis to derive new spatial indices. One hundred and five cross-sectional images from different species were transformed into binary images. The resulting point patterns, based on the locations of the conduit centers-of-area, were analyzed to determine whether they departed from randomness. Conduit distribution was then modeled using a spatially explicit stochastic model. * KEY RESULTS: The presence of conduit randomness, uniformity, or aggregation depended on the spatial scale of the analysis. The large majority of the images showed patterns significantly different from randomness at least at one spatial scale. A strong phylogenetic signal was detected in the spatial variables. * CONCLUSIONS: Conduit spatial arrangement has been largely conserved during evolution, especially at small spatial scales. Species in which conduits were aggregated in clusters had a lower conduit density compared to those with uniform distribution. Statistically sound spatial indices must be employed as an aid in the characterization of distributional patterns across species and in models of xylem water transport. Point pattern analysis is a very useful tool in identifying spatial patterns. PMID- 21616878 TI - Population biology of two rare fern species: long life and long-lasting stability. AB - PREMISE OF THE STUDY: This study describes the population dynamics of two rare fern species and evaluates the prospects of their survival. This is the first detailed demography study of ferns using transition matrix models. The study species, Asplenium adulterinum and A. cuneifolium, are restricted to serpentine rocks and differ in ploidy level and partly in habitat requirements. Both species are of interest in nature conservation. * METHODS: Single life-history traits were evaluated and transition matrix models were used to describe the dynamics of the populations. Population growth rates, elasticity values, and life-table response experiments were used to compare the dynamics between species, years, and different habitat types. Predicted population performance based on models was compared with real data on population growth. * KEY RESULTS: All populations of both species are growing. Stable stage distribution based on stochastic simulation corresponds to current stage distribution. The most critical phase of the life cycle is stasis of large adult plants. Reproduction is of low importance. Extinction probability of small populations is low. Mean life span of individuals of both species is 30-50 yr. When compared with real data, the model successfully predicted population performance over 10 yr. * CONCLUSION: Populations in the study region are not endangered, and current population dynamics are stable. Differences in life-history traits between species, probability of extinction between species and habitat, and different ploidy-and, thus, probably different dispersal ability-suggest the existence of metapopulation dynamics. PMID- 21616879 TI - Isotopic evidence of partial mycoheterotrophy in the Gentianaceae: Bartonia virginica and Obolaria virginica as case studies. AB - PREMISE OF THE STUDY: An estimated 10% of plant species have evolved to steal C from their symbiotic fungal partners (mycoheterotrophy), and while physiological evidence for full and partial mycoheterotrophy is well developed in the Orchidaceae and Ericaceae, it is lacking for the majority of other mycoheterotrophic taxa. The family Gentianaceae not only contains several lineages of achlorophyllous mycoheterotrophs, but also contains species that are putative partially mycoheterotrophic. The North American genera Bartonia and Obolaria (Gentianaceae) are green but have leaves reduced to scales or foliose bracts and so have ambiguous mycoheterotrophic status. * METHODS: We investigated the natural abundance (13)C and (15)N profiles of both genera along with total N and chlorophyll content and investigated mycorrhizal infection using light microscopy. * KEY RESULTS: The shoots of B. virginica were significantly more enriched in (15)N than the surrounding vegetation but not in (13)C. In contrast, the shoots of O. virginica are not enriched in (15)N compared to the surrounding vegetation but were significantly enriched in (13)C. Total N concentrations were significantly higher than the surrounding vegetation in B. virginica, while the collaroid roots of both species were infected by arbuscular mycorrhizal fungi. * CONCLUSIONS: This microscopic evidence coupled with the natural abundance stable isotope profiles strongly suggests that both species are partially mycoheterotrophic. However, differences in the root-shoot stable isotopic patterns relative to surrounding vegetation between B. virginica and O. virginica are suggestive of the utilization of different physiological pathways or extent of commitment to mycoheterotrophic C gain. PMID- 21616880 TI - A case study of modified interactions with symbionts in a hybrid mediterranean orchid. AB - PREMISE OF THE STUDY: Most studies on orchid hybrids examine separately the effects of hybridization on interactions with pollinators or with mycorrhizal fungi. Here, we simultaneously investigated both interactions in the mediterranean food-deceptive Orchis simia, O. anthropophora, and their hybrid (O. *bergonii) and tested a possible breakdown of coevolution using a multidisciplinary approach. * METHODS: We compared leaf growth, seed viability, emitted scent, and mycorrhizal fungi (species and rate of infection) among these three taxa. * KEY RESULTS: We show that leaf surface is greater in adult hybrids than in the parental species, suggesting a heterosis effect for vegetative growth. We demonstrate that flowers of the two parental species emit well differentiated bouquets of volatile organic compounds, while hybrids emit larger quantities, accumulating most compounds of the two parental species. However, hybrids fail to attract pollinators and have a 10 times lower fruit set. We determined that closely related Tulasnellales are mycorrhizal in the three taxa, suggesting that the mycorrhizal partner does not impair hybrid survival. We propose an interpretative model for O. *bergonii compared with its parents. * CONCLUSIONS: In hybrids, carbon resources normally devoted to reproduction may be reallocated to the mycorrhizal symbiosis as a result of the disruption of the pollination interaction in hybrids. Higher mycorrhizal infection may in turn enhance vegetative growth and scent emission. Such interplay between the two obligate biotic interactions yields new insights into hybridization among orchids. PMID- 21616881 TI - Species-specificity of nurse plants for the establishment, survivorship, and growth of a columnar cactus. AB - PREMISE OF THE STUDY: Seedling establishment and early survivorship are crucial steps for the regeneration of plant populations because both have long-lasting effects on plant population dynamics. For species recruiting through facilitation, species-specific facilitative effects might affect early fitness, an overlooked aspect in studies of facilitation considering groups of nurse species. * METHODS: We experimentally evaluated the roles of 10 nurse species and open space on the early performance of the columnar cactus Neobuxbaumia mezcalaensis. We measured establishment, survivorship, and growth of individuals over 3 years. Moreover, to study an extended period of the ontogeny of the interaction between this cactus and its nurse plants, we also monitored survivorship and growth rates of individuals between 3 to 12 cm tall during a 3 year period. * KEY RESULTS: Neobuxbaumia mezcalaensis performance varied significantly among nurse species, and only six yielded positive effects on early fitness. Densely canopied plants were the best nurses for this cactus. However, even among densely canopied species, some produced negative effects on the early fitness of N. mezcalaensis, indicating that similar nurse plants may elicit either facilitative or interference effects on beneficiary species. * CONCLUSIONS: Our results emphasize the importance of species-specific facilitative interactions in the crucial early stages in the life cycle of N. mezcalaensis and how different nurse species modify the effect of seed-rain and contribute significantly to the population dynamics of the species. PMID- 21616882 TI - The age and diversification of the angiosperms re-revisited. AB - PREMISE OF THE STUDY: It has been 8 years since the last comprehensive analysis of divergence times across the angiosperms. Given recent methodological improvements in estimating divergence times, refined understanding of relationships among major angiosperm lineages, and the immense interest in using large angiosperm phylogenies to investigate questions in ecology and comparative biology, new estimates of the ages of the major clades are badly needed. Improved estimations of divergence times will concomitantly improve our understanding of both the evolutionary history of the angiosperms and the patterns and processes that have led to this highly diverse clade. * METHODS: We simultaneously estimated the age of the angiosperms and the divergence times of key angiosperm lineages, using 36 calibration points for 567 taxa and a "relaxed clock" methodology that does not assume any correlation between rates, thus allowing for lineage-specific rate heterogeneity. * KEY RESULTS: Based on the analysis for which we set fossils to fit lognormal priors, we obtained an estimated age of the angiosperms of 167-199 Ma and the following age estimates for major angiosperm clades: Mesangiospermae (139-156 Ma); Gunneridae (109-139 Ma); Rosidae (108-121 Ma); Asteridae (101-119 Ma). * CONCLUSIONS: With the exception of the age of the angiosperms themselves, these age estimates are generally younger than other recent molecular estimates and very close to dates inferred from the fossil record. We also provide dates for all major angiosperm clades (including 45 orders and 335 families [208 stem group age only, 127 both stem and crown group ages], sensu APG III). Our analyses provide a new comprehensive source of reference dates for major angiosperm clades that we hope will be of broad utility. PMID- 21616883 TI - Ecological genetics of the Bromus tectorum (Poaceae)-Ustilago bullata (Ustilaginaceae) pathosystem: A role for frequency-dependent selection? AB - PREMISE OF THE STUDY: Evolutionary processes that maintain genetic diversity in plants are likely to include selection imposed by pathogens. Negative frequency dependent selection is a mechanism for maintenance of resistance polymorphism in plant-pathogen interactions. We explored whether such selection operates in the Bromus tectorum-Ustilago bullata pathosystem. Gene-for-gene relationships between resistance and avirulence loci have been demonstrated for this pathosystem. * METHODS: We used molecular markers and cross-inoculation trials to learn whether the SSR genotypes of the host exhibited resistance to co-occurring pathogen races, whether host genotypes within a population had equal disease probability, and whether a common resistance locus and its corresponding avirulence locus exhibited predicted allele frequency changes during an epidemic. * KEY RESULTS: Five of six putative resistance loci that conferred resistance to co-occurring pathogen races occurred in common host SSR genotypes. Some common genotypes within populations were more likely to be diseased than others, and genotype frequencies sometimes changed across years in patterns consistent with frequency dependent selection. Observed changes in frequency of resistance and virulence alleles during an epidemic provided further support, but evidence was inconclusive. * CONCLUSIONS: Frequency-dependent selection may operate at endemic disease levels in this pathosystem, but is difficult to detect because many susceptible plants escape infection. Most pathogen isolates were virulent on most host genotypes, minimizing the apparent importance of frequency-dependent selection even during epidemics. PMID- 21616884 TI - A narrow group of monophyletic Tulasnella (Tulasnellaceae) symbiont lineages are associated with multiple species of Chiloglottis (Orchidaceae): Implications for orchid diversity. AB - PREMISE OF THE STUDY: The Orchidaceae is characterized by exceptional species diversity. Obligate orchid mycorrhizae are predicted to determine orchid distributions, and highly specific relationships between orchids and fungi may drive orchid diversification. In this study, mycorrhizal diversity was examined in the terrestrial, photosynthetic orchid genus Chiloglottis to test the hypothesis of mycorrhizal-mediated diversification in the genus Chiloglottis. This orchid genus secures pollination by sexual deception, an obligate and highly specific pollination strategy. Here we asked whether the obligate orchid-fungal interactions are also specific. * METHODS: Two sequenced loci, the internal transcribed spacer region (ITS) and mitochondrial large subunit (mtLSU), were used to identify fungal isolates and assess fungal species diversity. Symbiotic germination of two species Chiloglottis aff. jeanesii and C. valida were used to assess germination potential of isolates and confirm mycorrhizal association. * KEY RESULTS: Phylogenetic analyses revealed that six representative Chiloglottis species spanning a broad survey of the genus were all associated with a narrow group of monophyletic Tulasnella fungal lineages. * CONCLUSIONS: The Chiloglottis Tulasnella interaction appears to be the first known case of such a narrow symbiont association across a broadly surveyed orchid genus. It appears that the specific pollination system of Chiloglottis, rather than specific orchid-fungal interactions has been the key driving force in the diversification of the genus. These findings also indicate that plant groups with highly specific mycorrhizal partners can have a widespread distribution. PMID- 21616885 TI - Embryonic inbreeding depression varies among populations and by mating system in Witheringia solanacea (Solanaceae). AB - PREMISE OF THE STUDY: Embryonic inbreeding depression is a key influence on mating system evolution and can be difficult to estimate in self-incompatible species. A pollen chase experiment was used to estimate the magnitude of embryonic inbreeding depression in Costa Rican Witheringia solanacea, a species polymorphic for self-incompatibility (SI). In a pollen chase experiment, bud self pollinations are followed after anthesis by outcross pollinations, with a comparable pair of outcross pollinations used as a control. Lowered seed set for the self-precedence treatment indicates embryonic inbreeding depression. * METHODS: Embryonic inbreeding depression was assayed for self-compatible (SC) individuals and for SI plants from two populations that differ quantitatively in the onset and enzymatic activity of their SI response. Microsatellite markers were used to assay the selfing rate of a sample of surviving progeny from the prior self-pollination treatment. * KEY RESULTS: SC individuals showed no evidence of embryonic inbreeding depression. In SI plants, prior self-pollination reduced seed number by 28-70%, depending on population. Microsatellite genotyping revealed that embryonic inbreeding depression was even more severe than estimated by the phenotypic data: for mature fruits resulting from self-pollination precedence, the majority of the progeny were the result of outcross fertilization. * CONCLUSIONS: Lineage-specific purging of recessive lethals has accompanied the evolution of SC in this species. SI populations show contrasting levels of embryonic inbreeding depression, with nearly complete embryonic lethality upon selfing in the Monteverde population. In the face of high embryonic inbreeding depression, an increase in selfing rate can evidently occur only under severe pollen limitation. PMID- 21616886 TI - Geographic variation in primary sex allocation per flower within and among 12 species of Pedicularis (Orobanchaceae): Proportional male investment increases with elevation. AB - PREMISE OF THE STUDY: The study of geographic variation in ecologically important traits within and among taxa is a first step toward understanding the environmental factors that contribute to population differentiation and species divergence. This study examines variation in mean sex allocation per flower (androecium mass/gynoecium mass) among 49 wild populations representing 12 Pedicularis species across an elevation gradient on the eastern Tibetan Plateau. * METHODS: We used population means to evaluate sources of variation in per flower sex allocation within and across species. In particular, we evaluate the relative influence of intrinsic (i.e., plant size, estimated as aboveground stem biomass) vs. extrinsic factors affecting mean sex allocation among populations. * KEY RESULTS: Mean sex allocation per flower (the relative investment in male floral organs) is negatively correlated with mean plant size; populations of large plants produce relatively female-biased flowers. This relationship between mean plant size and mean sex allocation is not statistically significant, however, when the effect of elevation is controlled statistically. Among populations within and across species, mean sex allocation increases with elevation. This relationship persists even when the effect of mean plant size is controlled statistically. Factors associated with increasing elevation appear to favor genotypes and/or taxa with male-biased flowers. * CONCLUSION: Extrinsic environmental conditions may be more important than intrinsic resource status in determining patterns of geographic variation in mean sex allocation among populations or species of Pedicularis. We cannot conclude whether the effect of elevation on mean sex allocation is the result of environmentally induced plasticity, genetically based adaptation, or species sorting, but it is only partly mediated by mean plant size. PMID- 21616887 TI - Molecular phylogeny and biogeographic diversification of Parthenocissus (Vitaceae) disjunct between Asia and North America. AB - PREMISE OF THE STUDY: Parthenocissus is a genus of the grape family Vitaceae and has a disjunct distribution in Asia and North America with members in both tropical and temperate regions. The monophyly of Parthenocissus has not yet been tested, and the species relationships and the evolution of its intercontinental disjunction have not been investigated with extensive sampling and molecular phylogenetic methods. * METHODS: Plastid (trnL-F, rps16, and atpB-rbcL) and nuclear GAI1 sequences of 56 accessions representing all 12 Parthenocissus species were analyzed with parsimony, likelihood, and Bayesian inference. Divergence times of disjunct lineages were estimated with relaxed Bayesian dating. Evolution of the leaflet number was assessed by tracing this character onto Bayesian trees using the Trace Character Over Trees option in the program Mesquite. * KEY RESULTS: Parthenocissus is monophyletic and sister to the newly described segregate genus Yua. Two major clades within Parthenocissus are recognizable corresponding to their distribution in Asia and North America. The disjunction between the two continents is estimated to be at 21.64 (95% higher posterior densities 10.23-34.89) million years ago. * CONCLUSIONS: Parthenocissus is likely to have derived from the Eocene boreotropical element. Its current Asian-North American disjunction is dated to the early Miocene, congruent with fossil and paleoclimatic evidence. The tropical species is nested within the temperate clade and is inferred to have dispersed from the adjacent temperate regions. Parthenocissus and Yua are best treated as distinct genera. Leaflet number in this genus has a complex history and cannot be used as a character for infrageneric classification. PMID- 21616888 TI - Molecular phylogeny, character evolution, and biogeography of the grammitid fern genus Lellingeria (Polypodiaceae). AB - PREMISE OF THE STUDY: The recognition of monophyletic genera for groups that have high levels of homoplastic morphological characters and/or conflicting results obtained by different studies can be difficult. Such is the case in the grammitid ferns, a clade within the Polypodiaceae. In this study, we aim to resolve relationships among four clades of grammitid ferns, which have been previously recovered either as a polytomy or with conflicting topologies, with the goal of circumscribing monophyletic genera. * METHODS: The sampling included 89 specimens representing 61 species, and sequences were obtained for two genes (atpB and rbcL) and four intergenic spacers (atpB-rbcL, rps4-trnS, trnG-trnR, and trnL-trnF), resulting in a matrix of 5091 characters. The combined data set was analyzed using parsimony, likelihood, and Bayesian methods. Ninety-six morphological characters were optimized onto the generated trees, using the parsimony method. * KEY RESULTS: Lellingeria is composed of two main clades, the L. myosuroides and the Lellingeria s.s. clades, which together are sister to Melpomene. Sister to all three of these is a clade with two species of the polyphyletic genus Terpsichore. In the L. myosuroides clade, several dispersal events occurred between the neotropics, Africa, and the Pacific Islands, whereas Lellingeria s.s. is restricted to the neotropics, with about 60% of its diversity in the Andes. * CONCLUSIONS: Overall, our results suggest that Lellingeria is monophyletic, with two clades that are easily characterized morphologically and biogeographically. Morphological characters describing the indument are the most important to define the clades within the ingroup. A small clade, previously considered in Terpsichore, should be recognized as a new genus. PMID- 21616889 TI - Phylogeography of the Japanese common sedge, Carex conica complex (Cyperaceae), based on chloroplast DNA sequence data and chromosomal variation. AB - PREMISE OF THE STUDY: Genetic and chromosomal variations in plants are often reflected in the geographical distribution patterns. Therefore, identifying such phylogeographical patterns on population is important for understanding the process of plant diversification and speciation, and analyzing both molecular and cytological aspects is necessary. * METHODS: We investigated the phylogeographic pattern and genetic diversity of the widespread Japanese sedge, Carex conica complex, based on chloroplast DNA haplotypes and chromosomal variations. * KEY RESULTS: Sixteen distinct haplotypes were detected from 258 individuals in the C. conica complex inferred from the cpDNA sequences of intergenic spacer (IGS) between atpB and rbcL, IGS between trnT and trnL, trnL intron, and IGS between trnL and trnF. Most haplotypes showed distinct geographical structures. Phylogenetic analyses revealed two major clades, clades I and II, among 16 haplotypes of the complex. The haplotypes of clade I were mainly found in eastern Japan, while most haplotypes of clade II were found in western Japan. Four intraspecific aneuploids of 2n = 32, 34, 36, and 38, with forming normal bivalents in meiotic divisions, were also found in the complex. The geographical distribution of these intraspecific aneuploids corresponded well with those of the haplotypes. * CONCLUSIONS: Our results suggest that the genetic diversity and chromosomal variations in the C. conica complex may have originated from contractions and expansions of geographical ranges affected by Quaternary climatic oscillations. PMID- 21616890 TI - Placing the woody tropical genera of Polygonaceae: A hypothesis of character evolution and phylogeny. AB - PREMISE OF THE STUDY: Taxonomic groups have often been recognized on the basis of geographic distinctions rather than accurately representing evolutionary relationships. This has been particularly true for temperate and tropical members from the same family. Polygonaceae exemplifies this problem, wherein the woody tropical genera were segregated from temperate members of the family and placed in the subfamily Polygonoideae as two tribes: Triplarideae and Coccolobeae. Modern phylogenetic studies, especially when inferred from many lines of evidence, can elucidate more probable hypotheses of relationships. This study builds on previous work in the family and aims to test the traditional classification of the tropical woody taxa, which have been understudied and undersampled compared to their temperate relatives. * METHODS: A phylogenetic study was undertaken with expanded sampling of the tropical genera with data from five plastid markers (psbA-trnH, psaI-accD, matK, ndhF, and rbcL), nuclear ribosomal DNA (ITS) and morphology. * KEY RESULTS: Results support the placement of nine of 12 genera of the Triplarideae and Coccolobeae within Eriogonoideae, in which these genera form a paraphyletic assemblage giving rise to Eriogoneae. The remaining woody tropical genera excluded from Eriogonoideae occur in the paleotropics. * CONCLUSIONS: Traditional characters used to delimit Coccolobeae and Triplarideae are not useful for defining monophyletic groups. The six-tepal condition is derived from the five-tepal condition, and unisexual flowers have arisen multiple times in different sexual systems. Ruminate endosperm has arisen multiple times in the family, suggesting this character is highly plastic. PMID- 21616891 TI - An evaluation of candidate plant DNA barcodes and assignment methods in diagnosing 29 species in the genus Agalinis (Orobanchaceae). AB - PREMISE OF THE STUDY: DNA barcoding has been proposed as a useful technique within many disciplines (e.g., conservation biology and forensics) for determining the taxonomic identity of a sample based on nucleotide similarity to samples of known taxonomy. Application of DNA barcoding to plants has primarily focused on evaluating the success of candidate barcodes across a broad spectrum of evolutionary divergence. Less attention has been paid to evaluating performance when distinguishing congeners or to differential success of analytical techniques despite the fact that the practical application and utility of barcoding hinges on the ability to distinguish closely related species. * METHODS: We tested the ability to distinguish among 92 samples representing 29 putative species in the genus Agalinis (Orobanchaceae) using 13 candidate barcodes and three analytical methods (i.e., threshold genetic distances, hierarchical tree-based, and diagnostic character differences). Due to questions regarding evolutionary distinctiveness of some taxa, we evaluated success under two taxonomic hypotheses. * KEY RESULTS: The psbA-trnH and trnT-trnL barcodes in conjunction with the "best close match" distance-based method best met the objectives of DNA barcoding. Success was also a function of the taxonomy used. * CONCLUSIONS: In addition to accurately identifying query sequences, our results showed that DNA barcoding is useful for detecting taxonomic uncertainty; determining whether erroneous taxonomy or incomplete lineage sorting is the cause requires additional information provided by traditional taxonomic approaches. The magnitude of differentiation within and among the Agalinis species sampled suggests that our results inform how DNA barcoding will perform among closely related species in other genera. PMID- 21616892 TI - Small variance in growth rates in annual plants has large effects on genetic drift. AB - PREMISE OF THE STUDY: Effective population size (N(e)) is a critical index of the evolutionary capacity of populations. Low N(e) indicates that standing genetic diversity is susceptible to loss via stochastic processes (and inbreeding) and is, therefore, unavailable for natural selection to act upon. Reported N(e) in plant populations is often quite low. What biological and ecological factors might produce such low N(e) * METHODS: We conducted a simulation model to test the effect of randomly assigned and autocorrelated growth rates of annual plants on plant-size distributions at the end of the growing season. Because plant size is directly correlated with reproductive output in annual plants, variation in plant size reflects variation in reproduction, and thus our modeled size distributions can be used to estimate N(e). * KEY RESULTS: Randomly assigned growth rates had a negligble effect on N(e)/N. Autocorrelated growth rates decreased N(e)/N as the length of the growing season increased. This was the case even when the variance in growth rates was as low as 0.1% of the mean. * CONCLUSIONS: While intrinsic plant biology can affect the degree of growth autocorrelation, ecological factors such as competition, herbivory, and abiotic stress can increase or decrease levels of growth autocorrelation. Ecological factors that increase growth autocorrelation can have significant effects on genetic drift within populations. PMID- 21616893 TI - Microsatellites for the mangrove tree Avicennia germinans (Acanthaceae): Tools for hybridization and mating system studies. AB - PREMISE OF THE STUDY: We developed a new set of microsatellite markers for the black mangrove Avicennia germinans, to provide new informative tools for further studies of the mating system, interspecific hybridization, and population genetics. * METHODS AND RESULTS: We used the microsatellite-enriched library approach to isolate and characterize 25 new primer pairs. Sixteen of them are polymorphic, showing a variable degree of variation in A. germinans, while nine were monomorphic in the samples examined. Eight exhibited private alleles in A. schaueriana. * CONCLUSIONS: These results indicate that these new microsatellite markers will be useful molecular tools for further studies of A. germinans and A. schaueriana population genetics, mating systems, and hybridization. PMID- 21616894 TI - New microsatellite markers for the rare plant Cercidiphyllum japonicum and their utility for Cercidiphyllum magnificum. AB - PREMISE OF THE STUDY: Microsatellite markers were developed for Cercidiphyllum japonicum to study population genetics of this endangered species native to both eastern China and Japan. * METHODS AND RESULTS: Using the Fast Isolation by AFLP of Sequences Containing Repeats (FIASCO) protocol, 12 microsatellite markers that were successfully amplified showed polymorphism when tested on 33 individuals from two populations in eastern China and Japan. Overall, the number of alleles per locus ranged between 5 and 18. Eleven markers could be easily amplified and were polymorphic in C. magnificum. * CONCLUSIONS: These results indicate that these microsatellite markers are adequate for detecting and characterizing population genetic structure in Cercidiphyllum at fine and range-wide geographical scales. PMID- 21616895 TI - Hygrochastic capsule dehiscence in New Zealand alpine Veronica (Plantaginaceae). AB - PREMISE OF THE STUDY: Plant movement is more widespread than often recognized, involving different organs and mechanisms. Hygrochasy (opening in response to moisture) is a capsule-opening movement that is widely believed to be predominantly a feature of plants of desert and arid zones, where it may protect against seed predators and harsh climatic conditions and restrict dispersal to favorable germination times and sites. However, recently it has been reported from a wider range of environments. This study demonstrates that hygrochasy is much more common than previously realized, extending the phenomenon to plants of alpine habitats. * METHODS: Capsules of 23 species of New Zealand Veronica were collected, and we used light microscopy to investigate the anatomy and biomechanics responsible for the opening mechanism. Additionally, we collected morphological data to identify common traits of hygrochastic species. * KEY RESULTS: Hygrochastic capsule dehiscence was found in 10 alpine Veronica species. The opening mechanism is based on an antagonistic reaction between a nonlignified swelling tissue and a lignified resistance tissue. In Veronica, hygrochasy is associated with erect, narrowly angustiseptate capsules on short peduncles of creeping subshrubs or cushion plants. * CONCLUSION: Hygrochasy is a common dehiscence type in New Zealand alpine Veronica, and for the first time, this mechanism is described in detail for plants in alpine habitats. We propose that hygrochasy provides an effective seed dispersal mechanism in solitary capsules embedded in cushion plants and may restrict dispersal within habitat patches. PMID- 21616896 TI - Merged trees in second-growth, fire-origin forests in Patagonia, Chile: positive spatial association patterns and their ecological implications. AB - * PREMISE OF THE STUDY: Negative density-dependent processes have been thought to be the primary cause of shifting spatial patterns of tree populations through time. The existence of adult tree clusters might challenge this classical prediction. Here, we document the prevalence of merged stems (clustering of mature trees leading to stem fusion) in second-growth forests of Nothofagus pumilio and hypothesize that it is nonrandom but predictable in space. * METHODS: We stem-mapped nine sites in second-growth edge and interior forests of fire origin and in mature forests of N. pumilio (>3500 trees) in central Patagonia, Chile. The spatial structure of stand-level and individual-level features was estimated with spatial analyses (pair-correlation function and nearest-neighbor distances). * KEY RESULTS: Multistemmed trees were merged clusters of separate individuals. Merged trees were predominantly found at the edge of the second growth forests. We found strong clustering (<=5 m) at forest edge sites and none at interior sites. Nearest-neighbor distance distributions were unimodal for unmerged trees and monotonically decreasing for merged trees; interstem distances were much smaller at the edge sites than at the interior sites. * CONCLUSIONS: The occurrence of merged trees at the forest edge, and the resulting high spatial aggregation of stems, is consistent with the hypothesis that establishment was probably aggregated. The spatial pattern found at the forest edge changes the standard spatial pattern sequence through time in temperate forests, altering traditional forest-stand-dynamics models. PMID- 21616897 TI - Changes in snowmelt date and summer precipitation affect the flowering phenology of Erythronium grandiflorum (glacier lily; Liliaceae). AB - PREMISE OF THE STUDY: Climate change has affected species worldwide, including alterations in phenology, migration patterns, distribution, and survival. Because Erythronium grandiflorum is an early-season bloomer, alterations in its phenology may have serious implications for many North American Rocky Mountain communities, including changes in resource availability for pollinators and herbivores. * METHODS: We investigated whether changes in the snowmelt date, summer temperature, and summer precipitation have altered the timing and abundance of flowering in E. grandiflorum by collecting long-term data on floral abundance from 1975-2008 in a series of 2 * 2 m plots at the Rocky Mountain Biological Laboratory (RMBL) in Gothic, Colorado in the United States. * KEY RESULTS: Snowmelt date and mean summer temperature were negatively correlated. Over the 30 yr study, the snowmelt date advanced by 4.14 d/decade, and mean summer temperature increased by 0.38 degrees C/decade. Summer precipitation was variable, showing no change. The first, peak, and last flowering dates of E. grandiflorum advanced an average of 3.2 d/decade. Furthermore, earlier snowmelt and greater summer precipitation in the previous year led to earlier flowering in E. grandiflorum. There was no change in flowering abundance in this species, indicating it may be controlled by a complex set of abiotic and biotic variables. * CONCLUSIONS: Our study indicates that snowmelt is arriving earlier at the RMBL, which has caused earlier flowering in E. grandiflorum. Because alterations in phenology can disrupt important ecological interactions, information on potential phenological shifts in species that interact with E. grandiflorum is essential in determining the net effect of climate-driven alterations in phenology. PMID- 21616898 TI - Salinity tolerance and genetic variability in freshwater and brackish Iris hexagona colonies. AB - PREMISE OF THE STUDY: Saltwater intrusion is one of the most widespread environmental threats to freshwater wetlands. Iris species worldwide are important members of these plant communities. Wetland irises reproduce clonally and sexually, which permits populations to spread and disperse in benign and stressful conditions. The ability of iris populations to tolerate and adapt to elevated salinity can play an important role in determining the long-term health of wetland ecosystems. * METHODS: We used microsatellite markers to evaluate population structure and genetic diversity, and we performed a common garden experiment to examine the effect of salinity on the growth and reproduction of wild Iris hexagona collected from freshwater and brackish wetlands. * KEY RESULTS: Colonies were genetically distinct, with average to high heterozygosity (0.55-0.66) for a clonal species. Salinity had negative linear effects on leaf mass (g), clonal growth (g), root mass (g), and flower numbers, and it had nonlinear effects on seed numbers and seed mass (mg). The greatest sexual reproduction occurred in the intermediate-salinity (4 parts per thousand) treatment. Flowering phenology was delayed for 5 days in the highest-salinity treatment. * CONCLUSIONS: We hypothesized that irises from brackish habitats would tolerate salinity better than freshwater irises would, but no difference in iris performance existed between the two habitats. The observed salinity tolerance and genetic diversity of I. hexagona indicate that populations will persist despite moderate increases in environmental salinity. PMID- 21616899 TI - Utility of a large, multigene plastid data set in inferring higher-order relationships in ferns and relatives (monilophytes). AB - PREMISE OF THE STUDY: The monilophytes (ferns and relatives)-the third largest group of land plants-exhibit a diverse array of vegetative and reproductive morphologies. Investigations into their early ecological and life-history diversification require accurate, well-corroborated phylogenetic estimates. We examined the utility of a large plastid-based data set in inferring backbone relationships for monilophytes. * METHODS: We recovered 17 plastid genes for exemplar taxa using published and new primers. We compared results from maximum likelihood and parsimony analyses, assessed the effects of removing rapidly evolving characters, and examined the extent to which our data corroborate or contradict the results of other studies, or resolve current ambiguities. * KEY RESULTS: Considering multifamily clades, we found bootstrap support comparable to or better than that in published studies that used fewer genes from fewer or more taxa. We firmly establish filmy ferns (Hymenophyllales) as the sister group of all leptosporangiates except Osmundaceae, resolving the second deepest split in leptosporangiate-fern phylogeny. A clade comprising Ophioglossaceae and Psilotaceae is currently accepted as the sister group of other monilophytes, but we recover Equisetum in this position. We also recover marattioid and leptosporangiate ferns as sister groups. Maximum-likelihood rate-class estimates are somewhat skewed when a long-branch lineage (Selaginella) is included, negatively affecting bootstrap support for early branches. * CONCLUSIONS: Our findings support the utility of this gene set in corroborating relationships found in previous studies, improving support, and resolving uncertainties in monilophyte phylogeny. Despite these advances, our results also underline the need for continued work on resolving the very earliest splits in monilophyte phylogeny. PMID- 21616900 TI - Origins and introgression of polyploid species in Mentzelia section Trachyphytum (Loasaceae). AB - PREMISE OF THE STUDY: Polyploid speciation has been important in plant evolution. However, the conditions that favor the origination and persistence of polyploids are still not well understood. Here, we examine origins of 16 polyploid species in Mentzelia section Trachyphytum. * METHODS: We used phylogeny reconstructions based on DNA sequences from plastid regions and the nuclear gene isocitrate dehydrogenase (idh) to construct hypotheses of introgression and polyploidization. * KEY RESULTS: Molecular data suggest that homoploid hybridization has been surprisingly common in Trachyphytum. Diploid species had unequal involvement in polyploid origins, but most polyploid taxa had allopolyploid origins from extant progenitors. A few polyploids with extreme phenotypes did not appear to have extant progenitors. We infer that the progenitors of these species were derived from extinct diploid lineages or ancestral lineages of multiple extant diploids. In agreement with other recent studies, we recovered molecular evidence of multiple phylogenetically distinct origins for several polyploid taxa, including the widespread octoploid M. albicaulis. * CONCLUSIONS: Evidence of high levels of introgression and allopolyploidy suggests that hybridization has played an important role in the evolution of Trachyphytum. Although idh sequences exhibited complicated evolution, including gene duplication, deletion, and recombination, they provided a higher percentage of informative characters for phylogeny reconstruction than the most variable plastid regions, allowing tests of hypotheses regarding polyploid origins. Given the necessity for rapidly evolving low-copy nuclear genes, researchers studying hybridization and polyploidy may increasingly turn to complex sequence data. PMID- 21616901 TI - Guyanagaster, a new wood-decaying sequestrate fungal genus related to Armillaria (Physalacriaceae, Agaricales, Basidiomycota). AB - PREMISE OF THE STUDY: Sequestrate basidiomycete fungi (e.g. "gasteromycetes") have foregone ballistospory and evolved alternative, often elaborate mechanisms of basidiospore dispersal with highly altered basidioma morphology. Sequestrate fungi have independently evolved in numerous Agaricomycete lineages, confounding taxonomic arrangements of these fungi for decades. Understanding the multiple origins and taxonomic affinities of sequestrate fungi provides insight into the evolutionary forces that can drastically alter basidioma morphology. In the neotropical rainforests of the Guiana Shield, we encountered a remarkable sequestrate fungus fruiting directly on decaying hardwood roots. The fungus' singular combination of traits include a wood-decaying habit; black, verrucose peridium; reduced stipe; and gelatinized basidiospore mass. * METHODS: Guyanagaster necrorhiza gen. et sp. nov. is described. Macro- and micromorphological characters were assessed and compared to most similar taxa. To determine the phylogenetic affinities of the fungus, DNA sequence data were obtained for the 18S, ITS, and 28S rDNA, RBP2, and EF1alpha regions and subjected to single- and multi-gene analyses. DNA sequences from fungal vegetative organs growing on decaying woody roots confirmed the wood-inhabiting lifestyle of Guyanagaster. * KEY RESULTS: Guyanagaster is morphologically unique among sequestrate fungi worldwide. Phylogenetic evidence places Guyanagaster in close relation to the wood-decaying mushroom genus Armillaria in the Physalacriaceae (Agaricales, Agaricomycetes, Basidiomycota). * CONCLUSIONS: Guyanagaster represents an independently evolved sequestrate form within the Physalacriaceae. Although molecular data confirm that Guyanagaster is closely related to Armillaria, the unusual features of this fungus suggest a case of radically divergent morphological evolution. PMID- 21616902 TI - Evolutionary and ecophysiological significance of sugar utilization by the peat moss Sphagnum compactum (Sphagnaceae) and the common charophycean associates Cylindrocystis brebissonii and Mougeotia sp. (Zygnemataceae). AB - PREMISE OF THE STUDY: The goal of this study was to illuminate the evolutionary history and ecological importance of plant mixotrophy-the uptake and utilization of exogenous organic compounds. * METHODS: We quantitatively assessed the effect of sugar amendments on laboratory growth of Sphagnum compactum as a representative emergent peat moss and two species of ecologically associated zygnematalean algae, Cylindrocystis brebissonii and Mougeotia sp. * KEY RESULTS: Together with observations published elsewhere, our results suggest that under carbon or light limitation, the uptake of exogenous sugars by cells of charophycean algae and peat mosses may help these organisms maintain positive carbon balance. Utilization of 1% glucose by aquatic-grown algae helped to relieve dissolved inorganic carbon limitation, enhancing photoautotrophic growth by factors of 9.0 and 1.7, respectively. After an 8-wk growth period, amendments of 1% and 2% glucose enhanced air-grown moss biomass by 28 and 39 times, respectively, that of controls lacking sugar amendments. After 9 wk, 1% fructose enhanced biomass by 21 times, and 2% sucrose enhanced biomass by 31 times. * CONCLUSION: Our results indicate that plant mixotrophy is an early-evolved trait. The results also indicate that quantitative differences in sugar utilization by bryophytes and charophycean algae correlate with relative investments in protective cell-wall polyphenolics measured in previous studies, suggesting that sugar utilization may subsidize the cost of producing phenolic wall compounds in bryophytes. PMID- 21616903 TI - Linkages among life-history components through time: Carryover effects on the demography of a short-lived polycarpic plant. AB - PREMISE OF THE STUDY: Current reproduction in polycarpic plants may be affected by a wide variety of factors, including carryover or "historical" effects derived from environments experienced early in life and from previous investments in organ preformation or other life functions, such as growth or reproduction. Historical effects as determinants of plant reproductive success in a specific season have received considerably less attention than events during the current reproductive episode, especially for short-lived plants. * METHODS: We used structural equation modeling to assess direct and indirect relations between past reproduction and both subsequent reproduction and growth in Helianthemum squamatum, a short-lived polycarpic plant. Additionally, we explored the effects of current reproduction on future survival (binomial variable) by using logistic regression. * KEY RESULTS: Historical effects derived from previous growth positively affected current reproduction, suggesting the existence of a reproductive hierarchy in the population, in which some individuals are consistently better growers and seeders. The lack of effects of current reproduction on future reproduction, together with the existence of a negative effect on future survival, suggests that to invest maximally in reproduction at the expense of a short life span may be the optimal strategy for H. squamatum, a species inhabiting very unpredictable ecosystems. * CONCLUSIONS: Historical effects derived from past investments in growth or reproduction may have important consequences for current plant performance and may play an essential role in shaping life histories. PMID- 21616904 TI - Spatiotemporal variation in the reproductive ecology of two parapatric subspecies of Oenothera cespitosa (Onagraceae). AB - * PREMISE OF THE STUDY: Flowering plants that rely on pollinators for most of their reproduction may experience unpredictable and inconsistent availability of effective pollinators throughout their reproductive lifetime. We investigated the reproductive ecology of two subspecies of the tufted evening primrose, Oenothera cespitosa, which occupy geographically and edaphically distinct habitats in western North America: O. cespitosa subsp. navajoensis inhabits sandstone soils on open sites or rocky slopes in the Colorado Plateau and O. cespitosa subsp. cespitosa grows in clay soils on talus slopes and exposed rocky ridges in the western Great Plains and northern Rocky Mountains of the United States. * METHODS: Pollen augmentation and selfing experiments, floral visitor observations, and single-visit effectiveness experiments were conducted over 4 years to examine the breeding system and spatiotemporal variation in pollinator behavior, assemblage, and abundance at different populations for each subspecies. * KEY RESULTS: Both subspecies of O. cespitosa were self-incompatible and pollen limited, suggesting that the relative abundance, effectiveness, and movement patterns of different insects as pollinators influenced the quality and quantity of seed production in these plants. Medium-sized vespertine hawkmoths (Hyles lineata, Sphinx vashti) were effective pollinators when present, as were large matinal bees (Anthophora affabilis, A. dammersi, Xylocopa tabaniformis androleuca), whereas small oligolectic Lasioglossum bees primarily functioned as pollen thieves in the evening and morning. * CONCLUSIONS: These findings highlight the importance of variability of pollinator composition and abundance in the evolution of plant breeding systems and reproductive success at varying spatial and temporal scales. PMID- 21616905 TI - Newly resolved relationships in an early land plant lineage: Bryophyta class Sphagnopsida (peat mosses). AB - PREMISE OF THE STUDY: The Sphagnopsida, an early-diverging lineage of mosses (phylum Bryophyta), are morphologically and ecologically unique and have profound impacts on global climate. The Sphagnopsida are currently classified in two genera, Sphagnum (peat mosses) with some 350-500 species and Ambuchanania with one species. An analysis of phylogenetic relationships among species and genera in the Sphagnopsida were conducted to resolve major lineages and relationships among species within the Sphagnopsida. * METHODS: Phylogenetic analyses of nucleotide sequences from the nuclear, plastid, and mitochondrial genomes (11 704 nucleotides total) were conducted and analyzed using maximum likelihood and Bayesian inference employing seven different substitution models of varying complexity. * KEY RESULTS: Phylogenetic analyses resolved three lineages within the Sphagnopsida: (1) Sphagnum sericeum, (2) S. inretortum plus Ambuchanania leucobryoides, and (3) all remaining species of Sphagnum. Sister group relationships among these three clades could not be resolved, but the phylogenetic results indicate that the highly divergent morphology of A. leucobryoides is derived within the Sphagnopsida rather than plesiomorphic. A new classification is proposed for class Sphagnopsida, with one order (Sphagnales), three families, and four genera. * CONCLUSIONS: The Sphagnopsida are an old lineage within the phylum Bryophyta, but the extant species of Sphagnum represent a relatively recent radiation. It is likely that additional species critical to understanding the evolution of peat mosses await discovery, especially in the southern hemisphere. PMID- 21616906 TI - A phylogeny and classification of the Muhlenbergiinae (Poaceae: Chloridoideae: Cynodonteae) based on plastid and nuclear DNA sequences. AB - PREMISE OF THE STUDY: To understand the origins of C(4) grasslands, we must have a better interpretation of plant traits via phylogenetic reconstruction. Muhlenbergiinae, the largest subtribe of C(4) grasses in Mexico and the southwestern United States (with 176 species), is taxonomically poorly understood. * METHODS: We conducted a phylogenetic analysis of 47 genera and 174 species using six plastid regions (ndhA intron, ndhF, rps16-trnK, rps16 intron, rps3, and rpl32-trnL) and the nuclear ITS 1 and 2 (ribosomal internal transcribed spacer) regions to infer evolutionary relationships and revise the classification. * KEY RESULTS: In our analyses, Muhlenbergia (ca. 153 species) is paraphyletic, with nine genera (Aegopogon, Bealia, Blepharoneuron, Chaboissaea, Lycurus, Muhlenbergia, Pereilema, Redfieldia, Schaffnerella, and Schedonnardus) found nested within. We recognized the following five well-supported monophyletic lineages within Muhlenbergia: subg. Muhlenbergia, with species that have phosphoenolpyruvate carboxykinase-like leaf anatomy and long, scaly rhizomes; subg. Trichochloa with long-lived species that are relatively tall (up to 3 m); subg. Clomena with 3-nerved upper glumes; sect. Pseudosporobolus species with narrow panicles and plumbeous spikelets; and sect. Bealia species with lemmas with hairy margins and midveins. * CONCLUSIONS: We propose expanding the circumscription of Muhlenbergia to include the other nine genera in this subtribe and make the following new combinations: Muhlenbergia subg. Bealia, M. diandra, M. geminiflora, M. paniculata, M. phleoides, M. subg. Pseudosporobolus (also lectotipified), M. solisii, M. tricholepis. We also propose several new names: M. ammophila, M. columbi, M. plumosa. Our phylograms suggest that Muhlenbergia originated in North America because the sister (Sohnsia filifolia and Scleropogoninae) is composed of predominantly North American species. PMID- 21616907 TI - Quantitative morphology and species delimitation under the general lineage concept: Optimization for Hedera (Araliaceae). AB - PREMISE OF THE STUDY: The use of continuous morphological characters in taxonomy is traditionally contingent on the existence of discrete diagnostic characters. When plant species are the result of recent divergence and gene flow and/or hybridization occur, the use of continuous morphological characters may help in species identification and delimitation. Between nine and 15 species have been recognized in the last treatments of Hedera. The recent divergence of the species and the involvement of allopolyploidization as the main force in this process may have greatly impeded the establishment of clear limits and contributed to multiple taxonomic proposals. * METHODS: A multivariate statistical decision making procedure was applied to 56 quantitative morphological characters and 602 specimens to identify and delimit Hedera species under the general lineage concept. Species' exclusive genetic ancestry was evaluated with the genealogical sorting index from the Bayesian inference trees of 30 Hedera ITS sequences. * KEY RESULTS: The decision-making procedure allowed recognizing 12 species and two groups (stellate and scale-like trichome groups) in Hedera and provided statistical support for making decisions about long-standing taxonomic controversies. Common ancestry was detected for the populations of three species even in the absence of the species monophyly. * CONCLUSIONS: Quantitative variation supports discrete variation and provides statistical support for the taxa recognized in some recent proposals of Hedera. The need of explicit analysis of quantitative data are claimed to reduce taxonomic subjectivity and ease decision-making when qualitative data fail. PMID- 21616908 TI - Clonal structure of wild populations and origins of horticultural stocks of Illicium parviflorum (Illiciaceae). AB - PREMISE OF THE STUDY: Habitat fragmentation is often assumed to result in limited genetic diversity across impacted plant communities. Central Florida has undergone extensive anthropogenic changes, while also harboring large numbers of endemic species. In this study, we assessed genetic structure and dependence on clonality in a central Florida endemic, Illicium parviflorum (Illiciaceae), as well as evaluated genetic diversity of this species in horticultural stocks. * METHODS: Six sites were sampled across the geographic range of I. parviflorum. A PCR-based assay using intersimple sequence repeats (ISSRs) was used to assess genetic structure. * KEY RESULTS: Results, based on 26 ISSR loci, suggest that clonal structure plays a role in all populations, with PD values ranging from 0.25 to 0.50. Only two populations exhibited unique genotypes, while the remaining four populations shared genotypes. Horticultural samples all shared one genotype, which can be traced back to a single natural population. * CONCLUSIONS: Clonal reproduction is an important factor in the maintenance of natural populations of I. parviflorum, although the degree to which this is true varies by population. Horticultural samples likely represent a single or very few collection events, indicating the need for greater genetic diversity within horticultural stocks. Further analyses using microsatellites are planned to confirm these results. PMID- 21616909 TI - Evidence of a high-Andean, mid-Holocene plant community: An ancient DNA analysis of glacially preserved remains. AB - PREMISE OF THE STUDY: Around the world, tropical glaciers and ice caps are retreating at unprecedented rates because of climate change. In at least one location, along the margin of the Quelccaya Ice Cap in southeastern Peru, ancient plant remains have been continually uncovered since 2002. We used genetic analysis to identify plants that existed at these sites during the mid-Holocene. * METHODS: We examined remains between 4576 and 5222 yr old, using PCR amplification, cloning, and sequencing of a fragment of the chloroplast trnL intron. We then matched these sequences to sequences in GenBank. * KEY RESULTS: We found evidence of at least five taxa characteristic of wetlands, which occur primarily at lower elevations in the region today. * CONCLUSIONS: A diverse community most likely existed at these locations the last time they were ice-free and thus has the potential to reestablish with time. This is the first genetic analysis of vegetation uncovered by receding glacial ice, and it may become one of many as ancient plant materials are newly uncovered in a changing climate. PMID- 21616910 TI - The mysterious evolutionary origin for the GNE gene and the root of bilateria. AB - Phylogenomic analyses have revealed several important metazoan clades, such as the Ecdysozoa and the Lophotrochozoa. However, the phylogenetic positions of a few taxa, such as ctenophores, chaetognaths, acoelomorphs, and Xenoturbella, remain contentious. Thus, the findings of qualitative markers or "rare genomic changes" seem ideal to independently test previous phylogenetic hypotheses. We here describe a rare genomic change, the presence of the gene UDP-GlcNAc 2 epimerase/N-acetylmannosamine kinase (GNE). We show that GNE is encoded in the genomes of deuterostomes, acoelomorphs and Xenoturbella, whereas it is absent in protostomes and nonbilaterians. Moreover, the GNE has a complex evolutionary origin involving unique lateral gene transfer events and/or extensive hidden paralogy for each protein domain. However, rather than using GNE as a phylogenetic character, we argue that rare genomic changes such as the one presented here should be used with caution. PMID- 21616911 TI - Long-range and targeted ectopic recombination between the two homeologous chromosomes 11 and 12 in Oryza species. AB - Whole genome duplication (WGD) and subsequent evolution of gene pairs have been shown to have shaped the present day genomes of most, if not all, plants and to have played an essential role in the evolution of many eukaryotic genomes. Analysis of the rice (Oryza sativa ssp. japonica) genome sequence suggested an ancestral WGD ~50-70 Ma common to all cereals and a segmental duplication between chromosomes 11 and 12 as recently as 5 Ma. More recent studies based on coding sequences have demonstrated that gene conversion is responsible for the high sequence conservation which suggested such a recent duplication. We previously showed that gene conversion has been a recurrent process throughout the Oryza genus and in closely related species and that orthologous duplicated regions are also highly conserved in other cereal genomes. We have extended these studies to compare megabase regions of genomic (coding and noncoding) sequences between two cultivated (O. sativa, Oryza glaberrima) and one wild (Oryza brachyantha) rice species using a novel approach of topological incongruency. The high levels of intraspecies conservation of both gene and nongene sequences, particularly in O. brachyantha, indicate long-range conversion events less than 4 Ma in all three species. These observations demonstrate megabase-scale conversion initiated within a highly rearranged region located at ~2.1 Mb from the chromosome termini and emphasize the importance of gene conversion in cereal genome evolution. PMID- 21616912 TI - Odor hedonic capacity and anhedonia in schizophrenia and unaffected first-degree relatives of schizophrenia patients. AB - OBJECTIVE: There is increasing evidence that schizophrenia patients have difficulties in the hedonic appraisal of odors. In a prior study, we assessed olfactory hedonic perception birhinally and found that males with schizophrenia failed to attach the appropriate hedonic valence to a pleasant odor, despite correctly perceiving changes in odor intensity. Female patients, in contrast, exhibited normal responses. The current study extends this work by examining odor valence processing in unaffected first-degree relatives of schizophrenia patients, to determine the extent to which this abnormality may be genetically mediated. We also examine odor valence processing unirhinally, rather than birhinally, to probe possible lateralized differences in patients' hedonic processing deficits. METHOD: Individuals with schizophrenia (n = 54), first degree unaffected family members (n = 22), and demographically matched controls (n = 45) were administered the Suprathreshold Amyl Acetate Odor Intensity and Odor Pleasantness Rating Test. RESULTS: In contrast to family members and controls, both male and female schizophrenia probands underevaluated the hedonic characteristics of amyl acetate at lower concentrations and overevaluated its pleasantness at concentrations perceived as unpleasant by both controls and relatives. These patient-specific differences could not be explained by differences in smoking habit, medication use, or subjective ratings of odor intensity. However, they were associated with increased levels of anhedonia/asociality and negative symptomatology. CONCLUSIONS: Our findings suggest that both male and female schizophrenia patients have difficulties in the unirhinal appraisal of hedonic valence. Normal responses in unaffected first degree relatives suggest that this is an environmentally, rather than genetically, mediated abnormality denoting negative symptomatology. PMID- 21616914 TI - Incidence of malignancy in patients treated for antineutrophil cytoplasm antibody associated vasculitis: follow-up data from European Vasculitis Study Group clinical trials. AB - OBJECTIVES: Because standard immunosuppressive treatment for antineutrophil cytoplasm antibody-associated vasculitis (AAV) (granulomatosis with polyangiitis (Wegener's) (GPA) and microscopic polyangiitis (MPA)) has been associated with a significant risk of developing cancer, the cancer incidence of treated AAV patients was assessed. METHODS: This analysis concerned 535 patients with newly diagnosed AAV from 15 countries who had been enrolled between 1995 and 2002 in four European clinical trials. Over the period 2004-7, study participants' follow up events were updated, including cancers diagnosed. Age, sex and area standardised incidence ratios (SIR) and their 95% CI were calculated by linkage to five national cancer databases. RESULTS: During the 2650 person-years' observation period, 50 cancers were diagnosed in 46 patients. SIR (95% CI) were 1.58 (1.17 to 2.08) for cancers at all sites, 1.30 (0.90 to 1.80) for cancers at all sites excluding non-melanoma skin cancer (NMSC), 2.41 (0.66 to 6.17) for bladder cancer, 3.23 (0.39 to 11.65) for leukaemia, 1.11 (0.03 to 6.19) for lymphoma and 2.78 (1.56 to 4.59) for NMSC. Subgroup SIR for cancers at all sites were 1.92 (1.31 to 2.71) for GPA and 1.20 (0.71 to 1.89) for MPA. CONCLUSIONS: Cancer rates for AAV patients treated with conventional immunosuppressive therapy exceeded those expected for the general population. This cancer excess was largely driven by an increased incidence of NMSC. The smaller cancer risk magnitude in this cohort, compared with previous studies, might reflect less extensive use of cyclophosphamide in current treatment protocols. Longer follow up data are warranted to appraise the risk of developing cancers later during the course of AAV. PMID- 21616913 TI - The Ile585Val TRPV1 variant is involved in risk of painful knee osteoarthritis. AB - OBJECTIVE: To assess if a coding variant in the gene encoding transient receptor potential cation channel, subfamily V, member 1 (TRPV1) is associated with genetic risk of painful knee osteoarthritis (OA). METHODS: The Ile585Val TRPV1 variant encoded by rs8065080 was genotyped in 3270 cases of symptomatic knee OA, 1098 cases of asymptomatic knee OA and 3852 controls from seven cohorts from the UK, the USA and Australia. The genetic association between the low-pain genotype Ile-Ile and risk of symptomatic and asymptomatic knee OA was assessed. RESULTS: The TRPV1 585 Ile-Ile genotype, reported to be associated with lower thermal pain sensitivity, was associated with a lower risk of symptomatic knee OA in a comparison of symptomatic cases with healthy controls, with an odds ratio (OR) of 0.75 (95% CI 0.64 to 0.88; p=0.00039 by meta-analysis) after adjustment for age, sex and body mass index. No difference was seen between asymptomatic OA cases and controls (OR=1.02, 95% CI 0.82 to 1.27 p=0.86) but the Ile-Ile genotype was associated with lower risk of symptomatic versus asymptomatic knee OA adjusting for covariates and radiographic severity (OR=0.73, 95% CI 0.57 to 0.94 p=0.0136). TRPV1 expression in articular cartilage was increased by inflammatory cytokines (tumour necrosis factor alpha and interleukin 1). However, there were no differences in TRPV1 expression in healthy and arthritic synovial tissue. CONCLUSIONS: A genotype involved in lower peripheral pain sensitivity is significantly associated with a decreased risk of painful knee OA. This indicates a role for the pro-nociceptive gene TRPV1 in genetic susceptibility to symptomatic knee OA, which may also be influenced by a role for this molecule in cartilage function. PMID- 21616915 TI - Calpain-7 binds to CHMP1B at its second alpha-helical region and forms a ternary complex with IST1. AB - Some intracellular proteins involved in the endosomal sorting complex required for transport (ESCRT) system have microtubule interacting and transport (MIT) domains and bind to ESCRT-III protein family members named charged multivesicular body proteins (CHMPs) at their C-terminal regions containing MIT-interacting motifs (MIMs). While two types of MIMs (MIM1 and MIM2) have been reported, CHMP1B has MIM1 and IST1 has both MIM1 and MIM2. Previously, we demonstrated that CHMP1B and IST1 directly interacted with a tandem repeat of MIT domains of calpain-7 (CL7MIT) and that autolytic activity of calpain-7 was enhanced by IST1 in vitro but not by overexpression of IST1 in HEK293T cells. In this study, we detected enhancement of autolysis of mGFP-fused calpain-7 by coexpression with CHMP1B and observed further activation by additional coexpression of IST1 in HEK293T cells. We found that CL7MIT interacted with the second alpha-helical region of CHMP1B but not with the canonical C-terminal region containing MIM1 in vitro. Co immunoprecipitation assays demonstrated that the interaction between CL7MIT and CHMP1B and between CL7MIT and IST1 became stronger when IST1 or CHMP1B was additionally coexpressed, suggesting formation of ternary complex of calpain-7, IST1 and CHMP1B. Moreover, subcellular fractionation analyses revealed increase of calpain-7 in membrane/organelle fractions by concomitant overexpression of these ESCRT-III family member proteins. PMID- 21616916 TI - AKT down-regulates insulin-like growth factor-1 receptor as a negative feedback. AB - As a member of receptor tyrosine kinase (RTK) family, insulin-like growth factor 1 (IGF1) receptor (IGF1R) activates several downstream pathways to transmit proliferative signals from extracellular stimulation. AKT as a major effector plays a pivotal role in integrating various survival signalling cascades. Our data here show that hyperactive AKT leads to the decrease of IGF1R at the transcriptional level, which could be partly restored by phosphatidylinositol-3 kinase (PI3K) inhibitors including wortmannin and LY294002. Moreover, the decrease of IGF1R impairs the sensitivity of IRS1 to the stimulation by IGF1. mTOR as a main downstream target of AKT is not involved in the AKT-mediated down regulation of IGF1R. PMID- 21616917 TI - Vascular function in the diagnostic categories of polycystic ovary syndrome. AB - BACKGROUND: Polycystic ovary syndrome (PCOS) is associated with increased risk of cardiovascular disease (CVD). However, it is unknown whether CVD risk is equivalent across the different diagnostic categories or phenotypes of PCOS, particularly in the new phenotypes that affect up to 50% of women with PCOS. Surrogate markers of CVD include endothelial function and arterial stiffness that are independent risk factors for CVD. The primary aim of this study was to assess whether milder phenotypes of PCOS have elevated CVD risk compared with controls. METHODS: This was a cross-sectional study, conducted in an academic medical centre, of overweight premenopausal women with either National Institute of Health (NIH) PCOS (n = 29) or milder non-NIH PCOS (n = 25) and controls without PCOS (n = 27). Primary outcomes were markers of vascular health, including endothelial function [peripheral arterial tonometry (PAT), asymmetric dimethylarginine (ADMA) and plasminogen activator inhibitor-1 (PAI-1)] and arterial stiffness [central pulse wave velocity (PWVc)]. Secondary outcomes were insulin resistance, glucose tolerance and inflammation (C-reactive protein). RESULTS: ADMA was significantly different across the three groups (P = 0.003). ADMA was similar for NIH and non-NIH PCOS (0.56 +/- 0.01 versus 0.53 +/- 0.02 umol/l, P = 1.0) and higher for both NIH PCOS compared with controls (0.56 +/- 0.01 versus 0.46 +/- 0.02 umol/l, P = 0.003) and non-NIH PCOS compared with controls (0.53 +/- 0.02 versus 0.46 +/- 0.02 umol/l, P = 0.046). PAI-1 (P = 0.20), PAT (P = 0.95) and PWVc (P = 0.67) were similar for the three groups. All results were maintained after adjustment for age and body mass index where significant differences in these potentially confounding factors occurred between groups. CONCLUSIONS: Women with NIH and non-NIH PCOS have elevated ADMA compared with controls independent of age and adiposity. This suggests that CVD risk, reflected by endothelial dysfunction, is increased in both traditional NIH and new milder non-NIH PCOS phenotypes. However, no differences in other markers of endothelial function or arterial stiffness were apparent between phenotypes in this PCOS cohort. PMID- 21616918 TI - Primary cT2 bladder cancer: a good candidate for radiotherapy combined with cisplatin for bladder preservation. AB - BACKGROUND: Bladder preservation therapy (BPT) has been attempted for patients with localized muscle-invasive bladder cancer. However, the indication for BPT has not yet been established. To identify patients who are good candidates for BPT, we evaluated our long-term experience with chemoradiation therapy (CRT) for bladder preservation. METHODS: Between 1994 and 2009, 82 patients with bladder cancer (clinical stage T2-N0M0) without concurrent upper urinary tract urothelial cancer were treated with CRT. Before CRT, the patients had a biopsy or resection of the tumor by transurethral resection (TUR). The response to CRT was evaluated by TUR, urine cytology and computed tomography. RESULTS: Thirty-two cases (39.0%) had a pathological complete response (pCR) that was defined as no microscopic residual tumor in the bladder. After TUR, 69 cases (84.0%) achieved local control of the cancer, which was considered as a clinical complete response (cCR). There was no significant association between achievement of pCR and examined parameters. The long-term results of CRT were evaluated in cCR cases. The median follow-up was 42.8 months (range, 4.1-155.1). The 5-year overall survival rate was 77.7% and 5-year progression-free survival rate was 64.5%. Clinical T stage and type of tumor (primary or recurrence) were predictive factors for overall survival as well as progression-free survival. In addition, primary cT2 cases had significantly better prognosis than cT3-4 and recurrent cases in overall survival and progression-free survival (P= 0.008 and P= 0.046, respectively). CONCLUSION: Cases with a primary cT2 tumor could be good candidates for BPT with radiation combined with cisplatin. PMID- 21616919 TI - Efficacy of continuous, daily, oral, ultra-low-dose 200 mg acyclovir to prevent herpes zoster events among bortezomib-treated patients: a report from retrospective study. AB - OBJECTIVE: Herpes zoster is the most common infection in patients treated with bortezomib-containing regimens for multiple myeloma. Some clinical trials have reported on the use of acyclovir prophylaxis to decrease the incidence of herpes zoster. However, the appropriate acyclovir dose and duration of prophylaxis remain unclear. The primary objective of this study was to evaluate the efficacy of continuous oral 200 mg/day acyclovir prophylaxis and the secondary objective was to determine the risk factors for developing herpes zoster. METHODS: We collected medical information from consecutive patients who received bortezomib with or without acyclovir prophylaxis for relapsed or refractory multiple myeloma at our hospital and retrospectively analyzed the efficacy of acyclovir prophylaxis and the parameters for predicting the risk factors for developing herpes zoster. The definition of acyclovir prophylaxis was oral continuous administration of 200 mg of once daily, without cessation, during the entire period of bortezomib treatment. RESULTS: Six of the 33 patients in the study developed herpes zoster during bortezomib treatment. No varicella-zoster virus reactivation was observed in the 19 patients in the acyclovir prophylaxis group. The incidence of herpes zoster was significantly higher in the group that did not receive acyclovir prophylaxis (43%, 6 of 14 patients) than in the group that did (0%, 0 of 19; P = 0.003). The predictive factors for varicella-zoster virus reactivation were male sex (P = 0.035) and the use of acyclovir (P = 0.003). CONCLUSIONS: Continuous prophylaxis by oral 200 mg/day acyclovir in multiple myeloma patients receiving bortezomib treatment is effective and sufficient in preventing herpes zoster. PMID- 21616920 TI - Polymorphism and ligand dependent changes in human glucagon-like peptide-1 receptor (GLP-1R) function: allosteric rescue of loss of function mutation. AB - The glucagon-like peptide-1 receptor (GLP-1R) is a key physiological regulator of insulin secretion and a major therapeutic target for the treatment of type II diabetes. However, regulation of GLP-1R function is complex with multiple endogenous peptides that interact with the receptor, including full-length (1-37) and truncated (7-37) forms of GLP-1 that can exist in an amidated form (GLP-1(1 36)NH2 and GLP-1(7-36)NH2) and the related peptide oxyntomodulin. In addition, the GLP-1R possesses exogenous agonists, including exendin-4, and the allosteric modulator, compound 2 (6,7-dichloro-2-methylsulfonyl-3-tert butylaminoquinoxaline). The complexity of this ligand-receptor system is further increased by the presence of several single nucleotide polymorphisms (SNPs) that are distributed across the receptor. We have investigated 10 GLP-1R SNPs, which were characterized in three physiologically relevant signaling pathways (cAMP accumulation, extracellular signal-regulated kinase 1/2 phosphorylation, and intracellular Ca2+ mobilization); ligand binding and cell surface receptor expression were also determined. We demonstrate both ligand- and pathway-specific effects for multiple SNPs, with the most dramatic effect observed for the Met149 receptor variant. At the Met149 variant, there was selective loss of peptide induced responses across all pathways examined, but preservation of response to the small molecule compound 2. In contrast, at the Cys333 variant, peptide responses were preserved but there was attenuated response to compound 2. Strikingly, the loss of peptide function at the Met149 receptor variant could be allosterically rescued by compound 2, providing proof-of-principle evidence that allosteric drugs could be used to treat patients with this loss of function variant. PMID- 21616921 TI - Ligand-dependent TrkA activity in brain differentially affects spatial learning and long-term memory. AB - In the central nervous system, the nerve growth factor (NGF) receptor TrkA is expressed primarily in cholinergic neurons that are implicated in spatial learning and memory, whereas the NGF receptor p75(NTR) is expressed in many neuronal populations and glia. We asked whether selective TrkA activation may have a different impact on learning, short-term memory, and long-term memory. We also asked whether TrkA activation might affect cognition differently in wild type mice versus mice with cognitive deficits due to transgenic overexpression of mutant amyloid-precursor protein (APP mice). Mice were treated with wild-type NGF (a ligand of TrkA and p75(NTR)) or with selective pharmacological agonists of TrkA that do not bind to p75(NTR). In APP mice, the selective TrkA agonists significantly improved learning and short-term memory. These improvements are associated with a reduction of soluble Abeta levels in the cortex and AKT activation in the cortex and hippocampus. However, this improved phenotype did not translate into improved long-term memory. In normal wild-type mice, none of the treatments affected learning or short-term memory, but a TrkA-selective agonist caused persistent deficits in long-term memory. The deficit in wild-type mice was associated temporally, in the hippocampus, with increased AKT activity, increased brain-derived neurotrophic factor precursor, increased neurotrophin receptor homolog-2 (p75-related protein), and long-term depression. Together, these data indicate that selective TrkA activation affects cognition but does so differently in impaired APP mice versus normal wild-type mice. Understanding mechanisms that govern learning and memory is important for better treatment of cognitive disorders. PMID- 21616923 TI - Event-related potentials to emotional stimuli in migrainous children. AB - The present preliminary study was aimed at investigating the electrocortical correlates of attentional allocation toward emotional stimuli in children and adolescents with migraine by means of the event-related potentials. The electroencephalogram was continuously recorded in 7 migrainous children and 8 healthy controls while they were looking at a series of pleasant, neutral, and unpleasant pictures. The mean amplitude of the Negative Central component of the event-related potentials was computed as an index of the allocation of attentional resources to the presented stimuli. Relative to controls, children with migraine displayed reduced fronto-central negativity and larger posterior positivity in response to emotional pictures. This effect was already evident, overall, in a time window preceding the Negative Central component. The smaller cortical negativity in response to emotional stimuli suggests reduced attentional engagement toward emotionally relevant stimuli, or might be interpreted in terms of advanced brain maturation in migraine children. PMID- 21616922 TI - The vascular effects of infection in Pediatric Stroke (VIPS) Study. AB - Understanding the vascular injury pathway is crucial to developing rational strategies for secondary stroke prevention in children. The multicenter Vascular Effects of Infection in Pediatric Stroke (VIPS) cohort study will test the hypotheses that (1) infection can lead to childhood arterial ischemic stroke by causing vascular injury and (2) resultant arteriopathy and inflammatory markers predict recurrent stroke. The authors are prospectively enrolling 480 children (aged 1 month through 18 years) with arterial ischemic stroke and collecting extensive infectious histories, blood and serum samples (and cerebrospinal fluid, when clinically obtained), and standardized brain and cerebrovascular imaging studies. Laboratory assays include serologies (acute and convalescent) and molecular assays for herpesviruses and levels of inflammatory markers. Participants are followed prospectively for recurrent ischemic events (minimum of 1 year). The analyses will measure association between markers of infection and cerebral arteriopathy and will assess whether cerebral arteriopathy and inflammatory markers predict recurrent stroke. PMID- 21616925 TI - Os odontoideum as a rare but possible complication in children with dyskinetic cerebral palsy: a clinical and neuroradiologic study. AB - The authors describe a 12-year-old boy with dyskinetic (athetoid-dystonic subtype) cerebral palsy and os odontoideum. Dystonic and choreoathetotic components in cerebral palsy are movement disorders that are difficult to treat and cause major disability. Dystonic posturing causes excessive flexion, extension, and rotation of the neck. Repetitive abnormal movements in patients with this type of cerebral palsy give rise to a higher incidence of pathologic conditions affecting the craniovertebral junction. Os odontoideum is one of these pathologies, and it represents a rare anomaly of the odontoid process. There are only a few reports describing os odontoideum in children with dyskinetic cerebral palsy. This clinical and neuroradiologic study focuses on the problem of atlantoaxial instability and os odontoideum in these forms of cerebral palsy, which is too often underestimated. PMID- 21616924 TI - Neurologic and neurobehavioral sequelae in children with human immunodeficiency virus (HIV-1) infection. AB - The range and extent of neurologic and neurobehavioral complications of human immunodeficiency virus (HIV-1) infection in children are under-described. Seventy eight children with HIV-1 infection (32 females) were assessed for neurologic complications. Forty-six children had abnormal neurology examinations. Thirty three children had global pyramidal tract signs, 5 had a hemiparesis, 4 had peripheral neuropathy, 18 had visual impairment, and 5 had hearing impairment. Thirty-nine of 63 children over 1 year of age had neurobehavioral problems. Of 24 children with HIV encephalopathy, 74% had severe immunosuppression and 45% were not receiving antiretroviral therapy. Twelve children had prior opportunistic central nervous system infections, and 9 had epilepsy. Diverse neurologic and neurobehavioral deficits are common in children with HIV-1 infection. Children with severe immunosuppression, who were not receiving antiretroviral therapy, were growth impaired and less than 1 year of age, were at greatest risk for developing neurologic complications. PMID- 21616926 TI - Interactive effects of elevated CO2 and drought on nocturnal water fluxes in Eucalyptus saligna. AB - Nocturnal water flux has been observed in trees under a variety of environmental conditions and can be a significant contributor to diel canopy water flux. Elevated atmospheric CO(2) (elevated [CO(2)]) can have an important effect on day time plant water fluxes, but it is not known whether it also affects nocturnal water fluxes. We examined the effects of elevated [CO(2)] on nocturnal water flux of field-grown Eucalyptus saligna trees using sap flux through the tree stem expressed on a sapwood area (J(s)) and leaf area (E(t)) basis. After 19 months growth under well-watered conditions, drought was imposed by withholding water for 5 months in the summer, ending with a rain event that restored soil moisture. Reductions in J(s) and E(t) were observed during the severe drought period in the dry treatment under elevated [CO(2)], but not during moderate- and post-drought periods. Elevated [CO(2)] affected night-time sap flux density which included the stem recharge period, called 'total night flux' (19:00 to 05:00, J(s,r)), but not during the post-recharge period, which primarily consisted of canopy transpiration (23:00 to 05:00, J(s,c)). Elevated [CO(2)] wet (EW) trees exhibited higher J(s,r) than ambient [CO(2)] wet trees (AW) indicating greater water flux in elevated [CO(2)] under well-watered conditions. However, under drought conditions, elevated [CO(2)] dry (ED) trees exhibited significantly lower J(s,r) than ambient [CO(2)] dry trees (AD), indicating less water flux during stem recharge under elevated [CO(2)]. J(s,c) did not differ between ambient and elevated [CO(2)]. Vapour pressure deficit (D) was clearly the major influence on night-time sap flux. D was positively correlated with J(s,r) and had its greatest impact on J(s,r) at high D in ambient [CO(2)]. Our results suggest that elevated [CO(2)] may reduce night-time water flux in E. saligna when soil water content is low and D is high. While elevated [CO(2)] affected J(s,r), it did not affect day time water flux in wet soil, suggesting that the responses of J(s,r) to environmental factors cannot be directly inferred from day-time patterns. Changes in J(s,r) are likely to influence pre-dawn leaf water potential, and plant responses to water stress. Nocturnal fluxes are clearly important for predicting effects of climate change on forest physiology and hydrology. PMID- 21616929 TI - Re: "Comparison of 2 approaches for determining the natural history risk of brain arteriovenous malformation rupture". PMID- 21616928 TI - Associations between general and abdominal adiposity and mortality in individuals with diabetes mellitus. AB - Individuals with diabetes mellitus are advised to achieve a healthy weight to prevent complications. However, fat mass distribution has hardly been investigated as a risk factor for diabetes complications. The authors studied associations between body mass index, waist circumference, waist/hip ratio, and waist/height ratio and mortality among individuals with diabetes mellitus. Within the European Prospective Investigation into Cancer and Nutrition, a subcohort was defined as 5,435 individuals with a confirmed self-report of diabetes mellitus at baseline in 1992-2000. Participants were aged 57.3 (standard deviation, 6.3) years, 54% were men, the median diabetes duration was 4.6 (interquartile range, 2.0-9.8) years, and 22% of the participants used insulin. Body mass index, as indicator of general obesity, was not associated with higher mortality, whereas all measurements of abdominal obesity showed a positive association. Associations generally were slightly weaker in women. The strongest association was observed for waist/height ratio: In the fifth quintile, the hazard rate ratio was 1.88 (95% confidence interval: 1.33, 2.65) for men and 2.46 (95% confidence interval: 1.46, 4.14) for women. Measurements of abdominal, but not general, adiposity were associated with higher mortality in diabetic individuals. The waist/height ratio showed the strongest association. Respective indicators might be investigated in risk prediction models. PMID- 21616930 TI - Degradation of glutathione S-conjugates in Physcomitrella patens is initiated by cleavage of glycine. AB - Glutathione-dependent detoxification is a key pathway that allows plants to efficiently remove toxic compounds like heavy metals or electrophilic xenobiotics. Under persistent exposure to toxins plants need to respond to continuous demand with efficient synthesis of glutathione (GSH) and ideally fast and efficient removal of potentially toxic glutathione S-conjugates. With the aim of studying the respective degradation pathway in Physcomitrella patens we initially characterized fluorescence labeling of protonema cultures with GSH specific xenobiotic monochlorobimane (MCB). Incubation of protonema with 200 MUM MCB for 24 h resulted in a steady increase of total bimane label, which was not confined to glutathione S-bimane (GS-B), but predominantly present in gamma glutamylcysteine S-bimane (gamma-EC-B) and cysteine S-bimane (Cys-B). Pulse-chase experiments identified gamma-EC-B and Cys-B as degradation products of GS-B, suggesting initial cleavage of the C-terminal glycine, followed by cleavage of the gamma-glutamyl bond. The amount of GS-B formed, increased linearly at 90 nmol GSH g fw-1 h-1 for 24 h and after ~1.5 h already surpassed the amount of GSH present in control protonema. This demand-driven biosynthesis of GSH depends on sufficient supply of sulfate in the incubation medium. PMID- 21616931 TI - Structure-function studies of an engineered scaffold protein derived from Stefin A. II: Development and applications of the SQT variant. AB - Constrained binding peptides (peptide aptamers) may serve as tools to explore protein conformations and disrupt protein-protein interactions. The quality of the protein scaffold, by which the binding peptide is constrained and presented, is of crucial importance. SQT (Stefin A Quadruple Mutant-Tracy) is our most recent development in the Stefin A-derived scaffold series. Stefin A naturally uses three surfaces to interact with its targets. SQT tolerates peptide insertions at all three positions. Peptide aptamers in the SQT scaffold can be expressed in bacterial, yeast and human cells, and displayed as a fusion to truncated pIII on phage. Peptides that bind to CDK2 can show improved binding in protein microarrays when presented by the SQT scaffold. Yeast two-hybrid libraries have been screened for binders to the POZ domain of BCL-6 and to a peptide derived from PBP2', specific to methicillin-resistant Staphylococcus aureus. Presentation of the Noxa BH3 helix by SQT allows specific interaction with Mcl-1 in human cells. Together, our results show that Stefin A-derived scaffolds, including SQT, can be used for a variety of applications in cellular and molecular biology. We will henceforth refer to Stefin A-derived engineered proteins as Scannins. PMID- 21616932 TI - Age-related differences in muscle fatigue vary by contraction type: a meta analysis. AB - BACKGROUND: During senescence, despite the loss of strength (force-generating capability) associated with sarcopenia, muscle endurance may improve for isometric contractions. PURPOSE: The purpose of this study was to perform a systematic meta-analysis of young versus older adults, considering likely moderators (ie, contraction type, joint, sex, activity level, and task intensity). DATA SOURCES: A 2-stage systematic review identified potential studies from PubMed, CINAHL, PEDro, EBSCOhost: ERIC, EBSCOhost: Sportdiscus, and The Cochrane Library. STUDY SELECTION: Studies reporting fatigue tasks (voluntary activation) performed at a relative intensity in both young (18-45 years of age) and old (>= 55 years of age) adults who were healthy were considered. DATA EXTRACTION: Sample size, mean and variance outcome data (ie, fatigue index or endurance time), joint, contraction type, task intensity (percentage of maximum), sex, and activity levels were extracted. DATA SYNTHESIS: Effect sizes were (1) computed for all data points; (2) subgrouped by contraction type, sex, joint or muscle group, intensity, or activity level; and (3) further subgrouped between contraction type and the remaining moderators. Out of 3,457 potential studies, 46 publications (with 78 distinct effect size data points) met all inclusion criteria. LIMITATIONS: A lack of available data limited subgroup analyses (ie, sex, intensity, joint), as did a disproportionate spread of data (most intensities >= 50% of maximum voluntary contraction). CONCLUSIONS: Overall, older adults were able to sustain relative-intensity tasks significantly longer or with less force decay than younger adults (effect size=0.49). However, this age related difference was present only for sustained and intermittent isometric contractions, whereas this age-related advantage was lost for dynamic tasks. When controlling for contraction type, the additional modifiers played minor roles. Identifying muscle endurance capabilities in the older adult may provide an avenue to improve functional capabilities, despite a clearly established decrement in peak torque. PMID- 21616933 TI - Measuring physical fitness in children who are 5 to 12 years old with a test battery that is functional and easy to administer. AB - BACKGROUND: Valid and reliable measures of children's physical fitness are necessary for investigating the relationship between children's physical fitness and children's health. Objective The objective of this study was to estimate the feasibility, internal consistency, convergent construct validity, and test-retest reliability of a new, functional, and easily administered test battery for measuring children's physical fitness. DESIGN: The study was a cross-sectional descriptive survey applying physical fitness tests across age groups 5 to 12 years. METHODS: Each of the 9 items in the test battery consists of a compound motor activity that recruits various combinations of endurance, strength (force generating capacity), agility, balance, and motor coordination: standing broad jump, jumping a distance of 7 m on 2 feet, jumping a distance of 7 m on one foot, throwing a tennis ball with one hand, pushing a medicine ball with 2 hands, climbing wall bars, performing a 10 * 5 m shuttle run, running 20 m as fast as possible, and performing a reduced Cooper test (6 minutes). The test battery was administered to 195 children (aged 5-12 years) from 4 schools and kindergartens in Norway. RESULTS: Overall, the children in each age group were able to perform all of the test items, indicating the suitability of the test battery for children as young as 5 years of age. With increasing age, total scores improved linearly, indicating the adequate sensitivity of the test battery for the age range examined in this study. Furthermore, even with the modest sample size used in this study, total scores were normally distributed, thereby fulfilling the necessary assumptions of most statistical procedures. For investigating the reliability of the test battery, 24 children (mean age=8.6 years) in one class were retested 1 week later. Test-retest correlations were high, with intraclass correlation coefficients for individual test items and total score ranging from .54 to .92. Limitations The survey was limited to samples of 5- to 12-year-old Norwegian children. Larger samples in each age group are essential for establishing age- and sex-specific norms. CONCLUSIONS: These promising results warrant further development of the test battery, including standardization and normalization based on a large, representative sample. PMID- 21616935 TI - Suppression of apoptosis by PIF1 helicase in human tumor cells. AB - Defining the processes that sustain telomere maintenance is critical to our understanding of cancer and longevity. PIF1 is a nonprocessive 5'->3' human DNA helicase that exhibits broad substrate specificity. In vitro studies have implicated PIF1 in maintaining telomeres and processing stalled DNA replication forks, but disruption of the murine Pif1 gene did not yield any apparent phenotype. In this study, we evaluated the function of the PIF1 gene in human cells by using siRNA knockdown strategies to gauge its role in the response to DNA replication stress. We found that PIF1 depletion reduced the survival of both p53-deficient and p53-proficient human tumor cells by triggering apoptosis. In contrast, nonmalignant cells were unaffected by PIF1 depletion. Apoptosis induction in tumor cells was augmented by cotreatment with replication inhibitors (thymidine, hydroxyurea, or gemcitabine). When sensitive PIF1-depleted cells were released from a thymidine-induced S-phase arrest, there remained a subpopulation of cells that failed to enter S-phase. This cell subpopulation displayed an increase in levels of cyclin E and p21, as well as a deficiency in S-phase checkpoint markers that were induced with thymidine in PIF1 expressing cells. Specifically, CHK1 activation was suppressed and we detected no consistent changes in ATM S1981 autophosphorylation, gammaH2AX induction, or RPA hyperphosphorylation. Death in PIF1-depleted cells was detected in late G(1)/early S-phase and was dependent on caspase-3 activity. Taken together, our findings suggest roles for PIF1 in S-phase entry and progression that are essential to protect human tumor cells from apoptosis. PMID- 21616934 TI - Associations between physical performance and executive function in older adults with mild cognitive impairment: gait speed and the timed "up & go" test. AB - BACKGROUND: Older adults with amnestic mild cognitive impairment (aMCI) are at higher risk for developing Alzheimer disease. Physical performance decline on gait and mobility tasks in conjunction with executive dysfunction has implications for accelerated functional decline, disability, and institutionalization in sedentary older adults with aMCI. OBJECTIVES: The purpose of this study was to examine whether performance on 2 tests commonly used by physical therapists (usual gait speed and Timed "Up & Go" Test [TUG]) are associated with performance on 2 neuropsychological tests of executive function (Trail Making Test, part B [TMT-B], and Stroop-Interference, calculated from the Stroop Word Color Test) in sedentary older adults with aMCI. DESIGN: The study was a cross-sectional analysis of 201 sedentary older adults with memory impairment participating in a longitudinal intervention study of cognitive function, aging, exercise, and health promotion. METHODS: Physical performance speed on gait and mobility tasks was measured via usual gait speed and the TUG (at fast pace). Executive function was measured with the TMT-B and Stroop Interference measures. RESULTS: Applying multiple linear regression, usual gait speed was associated with executive function on both the TMT-B (beta=-0.215, P=.003) and Stroop-Interference (beta=-0.195, P=.01) measures, indicating that slower usual gait speed was associated with lower executive function performance. Timed "Up & Go" Test scores (in logarithmic transformation) also were associated with executive function on both the TMT-B (beta=0.256, P<.001) and Stroop Interference (beta=0.228, P=.002) measures, indicating that a longer time on the TUG was associated with lower executive function performance. All associations remained statistically significant after adjusting for age, sex, depressive symptoms, medical comorbidity, and body mass index. LIMITATIONS: The cross sectional nature of this study does not allow for inferences of causation. CONCLUSIONS: Physical performance speed was associated with executive function after adjusting for age, sex, and age-related factors in sedentary older adults with aMCI. Further research is needed to determine mechanisms and early intervention strategies to slow functional decline. PMID- 21616936 TI - Preconditioned endothelial progenitor cells reduce formation of melanoma metastases through SPARC-driven cell-cell interactions and endocytosis. AB - Tumor progression is associated with the release of signaling substances from the primary tumor into the bloodstream. Tumor-derived cytokines are known to promote the mobilization and the recruitment of cells from the bone marrow, including endothelial progenitor cells (EPC). Here, we examined whether such paracrine influence could also influence the capacity of EPC to interfere with circulating metastatic cells. We therefore consecutively injected EPC prestimulated by tumor conditioned medium (EPC-CM) and luciferase-expressing B16 melanoma cells to mice. A net decrease in metastases spreading (vs. nonstimulated EPC) led us to carry out a 2-dimensional difference gel electrophoresis (2D-DIGE) proteomic study to identify possible mediators of EPC-driven protection. Among 33 proteins exhibiting significant changes in expression, secreted protein, acidic and rich in cysteine (SPARC) presented the highest induction after EPC exposure to CM. We then showed that contrary to control EPC, SPARC-silenced EPC were not able to reduce the extent of metastases when injected with B16 melanoma cells. Using adhesion tests and the hanging drop assay, we further documented that cell-cell interactions between EPC-CM and melanoma cells were promoted in a SPARC-dependent manner. This interaction led to the engulfment of melanoma cells by EPC-CM, a process prevented by SPARC silencing and mimicked by recombinant SPARC. Finally, we showed that contrary to melanoma cells, the prometastatic human breast cancer cell line MDA-MB231-D3H2 reduced SPARC expression in human EPC and stimulated metastases spreading. Our findings unravel the influence of tumor cells on EPC phenotypes through a SPARC-driven accentuation of macrophagic capacity associated with limitations to metastatic spread. PMID- 21616938 TI - RAD51C is a susceptibility gene for ovarian cancer. AB - A homozygous mutation in the RAD51C gene was recently found to cause Fanconi anemia-like disorder. Furthermore, six heterozygous deleterious RAD51C mutations were detected in German breast and ovarian cancer families. We screened 277 Finnish familial breast or ovarian cancer patients for RAD51C and identified two recurrent deleterious mutations (c.93delG and c.837+1G>A). These mutations were further genotyped in 491 familial breast cancer patients, 409 unselected ovarian cancer patients and two series of unselected breast cancer cases (884 from Helsinki and 686 from Tampere) and population controls (1279 and 807, respectively). The mutation frequency among all breast cancer cases was not different from the controls (4 out of 2239, 0.2% versus population controls 2 out of 2086, 0.1%, P= 0.7). In the Helsinki series, each mutation was found in four cases with personal or family history of ovarian cancer. No mutations were found among cases with familial breast cancer only, four out of the eight carriers did not have family history of breast cancer. The mutations associated with an increased risk of familial breast and ovarian cancer (OR: 13.59, 95% CI 1.89 97.6, P= 0.026 compared with controls), but especially with familial ovarian cancer in the absence of breast cancer (OR: 213, 95% CI 25.6-1769, P= 0.0002) and also with unselected ovarian cancer (OR: 6.31, 95% CI 1.15-34.6, P= 0.033), with a significantly higher mutation rate among the familial cases (two out of eight, 25%) than the unselected ovarian cancer cases (4 out of 409, 1%) (OR: 33.8, 95% CI 5.15-221, P= 0.005). These results suggest RAD51C as the first moderate-to high risk susceptibility gene for ovarian cancer. PMID- 21616937 TI - The mef/elf4 transcription factor fine tunes the DNA damage response. AB - The ATM kinase plays a critical role in initiating the DNA damage response that is triggered by genotoxic stresses capable of inducing DNA double-strand breaks. Here, we show that ELF4/MEF, a member of the ETS family of transcription factors, contributes to the persistence of gammaH2AX DNA damage foci and promotes the DNA damage response leading to the induction of apoptosis. Conversely, the absence of ELF4 promotes the faster repair of damaged DNA and more rapid disappearance of gammaH2AX foci in response to gamma-irradiation, leading to a radio-resistant phenotype despite normal ATM phosphorylation. Following gamma-irradiation, ATM phosphorylates ELF4, leading to its degradation; a mutant form of ELF4 that cannot be phosphorylated by ATM persists following gamma-irradiation, delaying the resolution of gammaH2AX foci and triggering an excessive DNA damage response. Thus, although ELF4 promotes the phosphorylation of H2AX by ATM, its activity must be dampened by ATM-dependent phosphorylation and degradation to avoid an excessive DNA damage response. PMID- 21616939 TI - Chronic exertional compartment syndrome. PMID- 21616940 TI - Quadratus lumborum asymmetry is not isolated to the dominant side in junior cricket fast bowlers. AB - BACKGROUND AND AIM: Bowling-side quadratus lumborum (QL) asymmetries have been previously reported on the dominant side in junior cricket fast bowlers using MRI. The aim of this study was to investigate QL asymmetry when measuring with two different methods; first using a small number of images with clear muscle borders and second using a larger number of images with less strict inclusion criteria. METHODS: MRI was performed on 38 junior (14.9 years) cricket fast bowlers prior to the start of a cricket season. Each MR image slice was evaluated to determine whether the QL muscle contour was visible and was assigned an image quality rating for inclusion in the study. The cross-sectional area of each included QL image was measured and compared with the corresponding image on the other side of the spine to determine side-to-side difference (asymmetries). RESULTS: Using the main method of including only high-quality MR images, 25% of MR images, where QL was in the field of view, met the inclusion criteria. The mean QL asymmetry was 13%, while 55% of participants had asymmetries greater than 10%. There was no significant difference in the number of participants with dominant and non-dominant side QL asymmetry. However, there was a significant difference in the magnitude of asymmetry between the dominant side (10.5%) and non-dominant (16.4%) asymmetries. The intraclass correlation coefficient for repeated measurements of QL asymmetry for randomly selected images (18%) was excellent (ICC 0.966, 95% CI 0.89 to 0.99). Using the second measurement method, with less strict inclusion criteria for MR images, similar results on the distribution of QL asymmetry were found. CONCLUSION: Contrary to previous research, this study demonstrated that there was a similar distribution of QL asymmetry between the dominant and non-dominant side. The presence of only dominant side asymmetry must therefore be questioned. PMID- 21616941 TI - Are anaesthetics toxic to the brain? AB - It has been assumed that anaesthetics have minimal or no persistent effects after emergence from anaesthesia. However, general anaesthetics act on multiple ion channels, receptors, and cell signalling systems in the central nervous system to produce anaesthesia, so it should come as no surprise that they also have non anaesthetic actions that range from beneficial to detrimental. Accumulating evidence is forcing the anaesthesia community to question the safety of general anaesthesia at the extremes of age. Preclinical data suggest that inhaled anaesthetics can have profound and long-lasting effects during key neurodevelopmental periods in neonatal animals by increasing neuronal cell death (apoptosis) and reducing neurogenesis. Clinical data remain conflicting on the significance of these laboratory data to the paediatric population. At the opposite extreme in age, elderly patients are recognized to be at an increased risk of postoperative cognitive dysfunction (POCD) with a well-recognized decline in cognitive function after surgery. The underlying mechanisms and the contribution of anaesthesia in particular to POCD remain unclear. Laboratory models suggest anaesthetic interactions with neurodegenerative mechanisms, such as those linked to the onset and progression of Alzheimer's disease, but their clinical relevance remains inconclusive. Prospective randomized clinical trials are underway to address the clinical significance of these findings, but there are major challenges in designing, executing, and interpreting such trials. It is unlikely that definitive clinical studies absolving general anaesthetics of neurotoxicity will become available in the near future, requiring clinicians to use careful judgement when using these profound neurodepressants in vulnerable patients. PMID- 21616942 TI - Is age a predictor of mortality in a U.K. medical high dependency unit? AB - BACKGROUND: The population aged older than 65 yr is set to increase by 32% by 2033. In resource-limited environments, difficult decisions regarding access to high dependency care for the elderly are increasingly important. The aim of this study was to determine whether age is a predictor of mortality in patients admitted to an open medical high dependency unit (MHDU). METHODS: Prospective observational cohort study of 100 consecutive patients admitted to an MHDU with a primary medical diagnosis over a 3 month period. The primary endpoint was 30 day mortality. RESULTS: Overall mortality at 30 days was 21% (n=21). Patients aged <65 yr were 41%, 29% were 65-74 yr, and 30% were aged 75 yr and above. There were no significant differences in mortality between groups (12%, 31%, and 23%, respectively). When considering Acute Physiology and Chronic Health Evaluation scores >=25, there was no significant difference in mortality between age groups [35% <70 yr (7/20) vs. 29% >=70 yr (4/14), P=1.000]. The final model at multivariable regression analysis identified that >=2 organ support (odds ratio 10.84, 95% confidence interval 3.28-35.84) and pre-admission moderate/nursing home care [4.44 (1.05-18.70)] were significantly associated with worse outcome. The majority of survivors (88%) were discharged at their pre-admission functional status; those who declined in function were not significantly older than those who did not. CONCLUSIONS: Age does not predict the outcome from an MHDU. Those requiring >=2 organ support, higher levels of pre-admission home support, or both had higher mortality. Selected elderly medical patients should not be denied high dependency unit care. PMID- 21616943 TI - First experience of monitoring with cardiac event recorder electrocardiography Omron system in childhood population for sporadic, potentially arrhythmia-related symptoms. AB - AIMS: To document symptomatic episodes of palpitations with traditional methods such as24 h Holter monitoring (HM) or loop recorders remains a big challenge in clinical practice. Clinical trials with patient-activated electrocardiography (ECG) recorders show increased diagnostic yield in such patients. However, studies in the paediatric population are limited. We want to present a first experience with an event-recording system Omron HeartScan in children with symptomatic palpitations. METHODS AND RESULTS: Thirty paediatric patients (age 4 16 years) were followed with the Omron at our centre. All patients had a normal echocardiogram, a normal baseline 12-lead ECG and a normal 24 h HM. Indications with regard to monitoring were palpitations (n = 30). Two of them also had episodes of pre-syncope. The average of palpitation episodes in the past 3 months was 13.2 +/- 8.3. The mean age of the study population was 9.7 +/- 2.3 years [17 males (56.7 %)]. In all patients (n = 30) a diagnostic event could be recorded with the studied system. Four patients were diagnosed with supraventricular tachycardia (SVT) and underwent catheter ablation. The remaining patients (n = 26; 87%) were diagnosed with sinus tachycardia. Two patients with SVT additionally had episodes of pre-syncope. None of the patients could be diagnosed with previous 24 h HM. CONCLUSION: This event recorder has a high diagnostic yield in the childhood population. The children enjoyed the ease of using the system under daily-life conditions. In this study the system was able to record a diagnostic event in all patients with palpitations. PMID- 21616944 TI - Clinical applications of magnets on cardiac rhythm management devices. AB - The growing indications for permanent pacemaker and implantable cardioverter defibrillator (ICD) implantation have increased the number of patients with these cardiac rhythm management devices (CRMDs). Cardiac rhythm management devices occasionally perform inappropriately in response to electromagnetic interference (e.g. surgical electrocautery) or lead noise over-sensing (e.g. lead fracture). Temporary reprogramming of the CRMDs using device programmers can prevent these untoward device responses. However, these programmers are device manufacturer specific and require technically qualified personnel to operate. This could cause delayed patient care and increased use of resources in certain clinical situations. Alternatively, clinical magnets, when appropriately positioned over the device site, can change the pacing to an asynchronous mode in pacemakers and suspend tachycardia therapies in ICDs. Although readily available, clinical magnets have not been widely used for this purpose, perhaps due to the unfamiliarity with the variable responses of CRMDs to magnet application. This article provides a comprehensive overview of the current literature on the mechanism of action and the specific responses of various CRMDs to clinical magnets. PMID- 21616945 TI - Patient-reported outcomes in Danish implantable cardioverter defibrillator patients with a Sprint Fidelis lead advisory notification. AB - AIMS: Few studies have investigated the association between implantable cardioverter defibrillators (ICDs) and lead advisory notifications and patient reported outcomes (PROs). We examined (i) whether the mode used to inform patients about a device advisory is associated with PROs, and (ii) whether patients with a lead subject to a device advisory report poorer PROs than non advisory controls. METHODS AND RESULTS: Patients (n= 207) implanted with an ICD at Aarhus University Hospital, Denmark, with a Sprint Fidelis lead subject to an advisory and a non-advisory control group (n= 510), completed a set of standardized PRO measures. A Bonferroni correction was applied to all statistical PRO comparisons to adjust for multiple comparisons, with a P-value of 0.0038 (0.05/13 PROs) indicating statistical significance. Device advisory patients did not differ significantly on PROs according to mode of notification (all P-values >0.0038). They also did not differ significantly from controls on mean scores of depression, anxiety, device acceptance, and health status (all P > 0.0038). Differences were only found on ICD concerns (P< 0.0001) and on mental health status (P = 0.003), with advisory patients reporting fewer ICD concerns and a better mental health status than non-advisory controls. CONCLUSIONS: The mode used to inform ICD patients about the advisory was not associated with PROs, nor was the overall well-being of device advisory patients impaired compared to non advisory controls. These results indicate that ICD patients are generally able to cope with a device advisory. PMID- 21616947 TI - Letter from Russia: alcoholism and dissent-report of a whistleblower. PMID- 21616946 TI - Urinary ethyl glucuronide and ethyl sulfate testing for recent drinking in alcohol-dependent outpatients treated with acamprosate or placebo. AB - AIMS: Ethyl glucuronide (EtG) and ethyl sulfate (EtS) are sensitive and specific biomarkers for recent alcohol ingestion. This study compared urinary EtG and EtS measurement with self-reports for detection of prior drinking in alcohol dependent outpatients treated with the anti-craving medication acamprosate or placebo. METHODS: Alcohol-dependent outpatients (26 women, 30 men) were randomized to 21 days of oral acamprosate (2 g/day) or placebo treatment in a double-blind design. They were instructed to refrain from drinking during the study. Return visits to the ward for blood and urine sampling and filling out questionnaires were made on Day 7, 14 (urine sample optional) and 22 (urine sample mandatory). EtG and EtS were determined by liquid chromatography-mass spectrometry. RESULTS: On the first day (Day 0), 72% of all patients (acamprosate 65%, placebo 78%) tested positive for recent drinking according to urinary EtG (reporting limit >= 0.5 mg/l) and EtS (>= 0.1 mg/l). On the final day (Day 22), the frequency of positive tests was significantly reduced to 30% in the acamprosate group (P = 0.0374) and 33% for placebo (P = 0.0050). However, there was no difference between the treatment groups. When both groups were combined, the EtG (P = 0.025) and EtS (P = 0.015) concentrations were lower on the final day. Altogether, EtG and EtS were detected in 76 of 156 (49%) urine samples. When drinking in the day before sampling was admitted, 93% of urines tested positive; when drinking was denied, still 28% of the samples were positive. CONCLUSION: These results confirmed the value of urinary EtG and EtS as reliable indicators of recent drinking during outpatient treatment of persons with alcohol-related problems, and as objective outcome measures when evaluating new alcohol treatment strategies and pharmacotherapies. PMID- 21616948 TI - Cnidoscolus aconitifolius leaf extract protects against hepatic damage induced by chronic ethanol administration in Wistar rats. AB - AIMS: This study focused on the possible protective effect of Cnidoscolus aconitifolius leaf extract (CA) against hepatic damage induced by chronic ethanol administration in rats. METHODS: Male Wistar rats were distributed into seven groups of six rats each. The first group was the control, second group received 20% ethanol-only (7.9 g/kg), third and fourth groups were pre-treated with CA (100 and 200 mg/kg, respectively) before treatment with ethanol. The fifth and sixth groups received CA and kolaviron (KV; 200 mg/kg), respectively, while the seventh group received KV and ethanol. KV served as the reference antioxidant. RESULTS: Ethanol-treated rats had significantly (P < 0.05) elevated serum and liver post-mitochondrial malondialdehyde, an index of lipid peroxidation. Ethanol toxicity lowered the antioxidant defense indices, such as reduced glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT). Specifically, the activities of hepatic SOD and CAT decreased by 48 and 51%, respectively, while the level of GSH decreased by 56%. In addition, serum total cholesterol, triglycerides and low-density lipoproteins-cholesterol levels were significantly (P < 0.05) elevated in ethanol-treated rats. Also, significant (P < 0.05) elevation in serum alanine and aspartate aminotransferases, and gamma-glutamyl transferase activities were observed in ethanol-treated rats. Supplementation with CA significantly (P < 0.05) decreased the activities of liver marker enzymes, stabilized the lipid profiles and restored the antioxidants status of ethanol-treated rats. The activities of CA were comparable with KV in the ethanol treated rats. This observation was supported by histopathological examination of liver slides. CONCLUSION: These findings suggest the hepatoprotective and antioxidant effects of CA leaf extract, which offered protection against ethanol induced toxicity. PMID- 21616949 TI - Bringing prevention to the population: an important role for cardiologists in policy-making. PMID- 21616950 TI - Early detection of pulmonary vascular disease in pulmonary arterial hypertension: time to move forward. AB - Pulmonary arterial hypertension (PAH) can be a rapidly progressive disorder and is associated with high rate of mortality, despite medical intervention. With the availability of effective therapy, early disease detection is an important strategic objective to improve treatment outcomes. Resting echocardiography is currently the recommended screening modality for high-risk population groups. However, it is clear that derangements in resting haemodynamics (and symptoms) are late sequelae of the pathobiological processes that begin in the distal pulmonary arteries. Exercise stress may unmask early pulmonary vascular dysfunction but the definition, clinical significance, and natural history of 'exercise PAH' remain undefined. We will review the currently available and potential future strategies aimed at early disease detection, and propose that ultimately the way forward is to detect disease at a stage prior to the rise in resting pulmonary artery pressure. PMID- 21616951 TI - Anticoagulation control in Sweden: reports of time in therapeutic range, major bleeding, and thrombo-embolic complications from the national quality registry AuriculA. AB - AIMS: In anticoagulation treatment with warfarin, the risk of thrombo-embolic events must be weighed against the risk of bleeding. Time in therapeutic range (TTR) is an important tool to assess the quality of anticoagulation treatment, and has been shown to correlate with less bleeding and thrombo-embolic complications. AuriculA, the Swedish national quality registry for atrial fibrillation and anticoagulation, is used for follow-up and dosage control of warfarin. This is the first report of TTR in AuriculA and, in a subgroup of two centres, bleeding and thrombo-embolic complications during 2008. METHODS AND RESULTS: Prothrombin complex (International normalized ratio) values from 18 391 patients in 67 different centres were analysed. The mean (SD) age was 70 (12) years. The main indications for warfarin treatment were: atrial fibrillation (64%), venous thromboembolism (19%), and heart valve dysfunction (13%). Time in therapeutic range for all patients was 76.2%. The mean weekly dose of warfarin decreased with age and TTR increased with age. In 4273 patients from two centres in AuriculA, the frequency of major bleedings and venous/arterial thrombo embolism were 2.6 and 1.7% and for atrial fibrillation, 2.6 and 1.4%, per treatment year, respectively. A correlation between age and the risk of major bleeding (P< 0.001), but not thrombo-embolic complications (P= 0.147), was seen. CONCLUSION: Compared with prospective randomized trials of warfarin treatment, TTR in the AuriculA population was higher. Complications were low, probably due to the organization of anticoagulation treatment in Sweden. Use of the AuriculA dosing programme could have contributed to the results by keeping dosing regimens consistent over all centres. PMID- 21616952 TI - The WHICH? trial: rationale and design of a pragmatic randomized, multicentre comparison of home- vs. clinic-based management of chronic heart failure patients. AB - AIMS: To describe the rationale and design of the Which Heart failure Intervention is most Cost-effective & consumer friendly in reducing Hospital care (WHICH?) trial. METHODS: WHICH? is a pragmatic, multicentre, randomized controlled trial that seeks to determine if multidisciplinary management of chronic heart failure (CHF) patients post-acute hospitalization delivered in a patient's own home is superior to care delivered via a specialist CHF outpatient clinic. The composite primary endpoint is all-cause, unplanned recurrent hospitalization or death during 12-18 months of follow-up. Of 688 eligible patients, 280 patients (73% male and 66% principal diagnosis of CHF) with a mean age of 71 +/- 14 years have been randomized to home- (n = 143) or clinic-based (n = 137) post-discharge management. This will provide 80% power (two-sided alpha of 0.05) to detect a 15% absolute difference in both the primary end-point and rate of all-cause hospital stay. Preliminary data suggest that the two groups are well matched in nearly all baseline socio-economic and clinical parameters. The majority of patients have significant co-morbidity, including hypertension (63%), coronary artery disease (55%), and atrial fibrillation (53%) with an accordingly high Charlson Index of Comorbidity Score (6.1 +/- 2.4). PERSPECTIVE: Despite its relatively small size, the WHICH? trial is well placed to examine the relative impact of two of the most commonly applied forms of face-to-face management designed to reduce recurrent hospitalization and prolong survival in CHF patients. PMID- 21616953 TI - Complementary role of copeptin and high-sensitivity troponin in predicting outcome in patients with stable chronic heart failure. AB - AIMS: Copeptin, the C-terminal part of the vasopressin pro-hormone, is elevated after myocardial infarction and predicts adverse outcome. In the present study we investigated whether the complementary role of copeptin and cardiac troponin T (cTnT) could be used for identification of high-risk patients with chronic stable heart failure. METHODS AND RESULTS: We measured copeptin and high-sensitivity cTnT (hs-cTnT) levels in 172 consecutive patients with stable chronic heart failure. Patients were followed for all-cause mortality and hospitalization due to heart failure for a median of 1301 days (interquartile range: 707-1636). In univariate analysis, plasma copeptin showed a moderate but significant correlation with hs-cTnT (r= 0.40 P< 0.001), age (r= 0.36 P< 0.001), creatinine (r= 0.52 P< 0.001), and amino-terminal pro-B type natriuretic peptide (NT-proBNP; r= 0.42 P< 0.001). Both copeptin (P= 0.002) and hs-cTnT (P= 0.005) concentrations were significantly increased in patients with higher New York Heart Association classes. While 109 (58%) patients had hs-cTnT concentrations above normal (>14 pg/mL) 104 patients (55%) had copeptin concentrations above normal (16.4 pmol/L). In survival analysis, both elevated copeptin and hs-cTnT concentrations were significant predictors of outcome (P< 0.001 for both). The combination of both markers showed a graded and highly significant association with impaired clinical outcome, which was independent of plasma NT-proBNP levels (adjusted hazard ratios 1.40, 95% CI, 1.20-1.70; P< 0.001). Adding copeptin concentrations to a prediction model with NT-proBNP and hs-cTnT resulted in significant improvement in model performance (net reclassification improvement 0.208; P< 0.05). CONCLUSION: Our data suggest that the combined use of hs-cTnT and copeptin might predict clinical outcome of patients with chronic stable heart failure. PMID- 21616954 TI - A systematic review of comprehensive geriatric assessment to improve outcomes for frail older people being rapidly discharged from acute hospital: 'interface geriatrics'. AB - BACKGROUND: many frail older people who attend acute hospital settings and who are discharged home within short periods (up to 72 h) have poor outcomes. This review assessed the role of comprehensive geriatric assessment (CGA) for such people. METHODS: standard bibliographic databases were searched for high-quality randomised controlled trials (RCTs) of CGA in this setting. When appropriate, intervention effects were presented as rate ratios with 95% confidence intervals. RESULTS: five trials of sufficient quality were included. There was no clear evidence of benefit for CGA interventions in this population in terms of mortality [RR 0.92 (95% CI 0.55-1.52)] or readmissions [RR 0.95 (95% CI 0.83 1.08)] or for subsequent institutionalisation, functional ability, quality-of life or cognition. CONCLUSIONS: there is no clear evidence of benefit for CGA interventions in frail older people being discharged from emergency departments or acute medical units. However, few such trials have been carried out and their overall quality was poor. Further well designed trials are justified. PMID- 21616955 TI - Embedding the Mental Capacity Act into clinical practice in England and Wales. PMID- 21616956 TI - Lifestyle behaviours and quality-adjusted life years in middle and older age. AB - OBJECTIVE: to examine the relationship between combined lifestyle behaviours and quality-adjusted life years (QALYs) in a general population. METHODS: a population-based study was conducted in 13,358 men and women who participated in the European Prospective Investigation into Cancer (EPIC)-Norfolk (baseline 1993 97). A score of 1 was given to each of non-smoking, physically not inactive, moderate alcohol consumption (1-14 units) and consumption of at least five portions of fruit and vegetables (vitamin C level >=50 umol/l). Short-Form Six Dimension (SF-6D) health utility index scores were derived from the SF-36. QALYs were estimated up to follow-up (July 2007). RESULTS: a total of 13,358 men and women were eligible to be included in the study (aged 40-79 years at baseline). A total of 12,921 people were alive at follow-up (117, 784 person-years). Mean follow-up period was ~11.5 years. 437 (4.4% of men and 2.4% of women) died. The death rate was 6.5 times higher in people with health behaviour score 0 compared with those who scored 4 (8.4 versus 1.3%). People with higher scores had significantly higher QALYs. CONCLUSION: our findings support the view that modifiable lifestyle factors are an important component in health improvement. PMID- 21616957 TI - Narrative review: the management of acute decompensated heart failure. AB - Acute decompensated heart failure (ADHF) is the most common reason for hospitalization in Western nations. The prognosis of patients admitted to hospital with ADHF is poor, with up to 64% being readmitted within the first 90 days after discharge and with a 1-year mortality approximating 20%. Epidemiological studies suggest that the majority of patients hospitalized with ADHF receive treatment that is inadequate and which is not based on scientific evidence. Furthermore, emerging data suggest that the "conventional" therapeutic interventions for ADHF including morphine, high-dose diuretics, and inotropic agents may be harmful. The goal of this review is to provide evidence-based recommendations for the diagnosis and management of ADHF. PMID- 21616958 TI - Ischaemic stroke; not just a disease of adulthood. PMID- 21616959 TI - A patient with suspicious violaceous nodules. PMID- 21616960 TI - In utero programming of pubertal development? PMID- 21616961 TI - Feedback on doctors' performance from parents and carers of children: a national pilot study. AB - OBJECTIVES: To evaluate the reliability and validity of a children's carers' feedback tool, to explore the feasibility of delivering this nationally and to determine acceptability to doctors of this assessment. PARTICIPANTS: 122 UK paediatricians on the specialist register undertaking outpatient consultations. DESIGN: Participants were each sent 50 forms for distribution to carers. Mean scores for each question, and for the overall pilot cohort were returned to participants with verbatim free text comments. Participating paediatricians' views were sought before and after receiving feedback. RESULTS: 122 doctors returned 4415 forms (mean 36 per doctor). All doctors scored highly with scores across all returned forms having a median of 4.58 (IQ range 0.17) where the maximum score was 5. Differences were observed between scores from female compared to male carers (p<0.05), from consultations rated by carer and child compared to carer alone (p<0.05) and from carers who had previously met the doctor compared to those in their first consultation (p<0.001). 'White' doctors received higher ratings than 'non-white' doctors (p<0.05) and white patients rated both white doctors and non-white doctors more highly than non-white patients (p<0.01). A minimum of 25 consultations rated by children's carers are needed for acceptable reliability. 93.9% of participants would be happy to be assessed in this way for the purposes of revalidation. CONCLUSIONS: National delivery of a valid and reliable method of carer feedback is feasible. The scores received and acceptability in these self-selected doctors was high. Confounding variables may influence feedback, so guidance on interpretation may be needed. PMID- 21616962 TI - A comment on impaired peri-nidal cerebrovascular reserve in seizure patients with brain arteriovenous malformations. PMID- 21616963 TI - The contribution of the putamen to sensory aspects of pain: insights from structural connectivity and brain lesions. AB - Cerebral cortical activity is heavily influenced by interactions with the basal ganglia. These interactions occur via cortico-basal ganglia-thalamo-cortical loops. The putamen is one of the major sites of cortical input into basal ganglia loops and is frequently activated during pain. This activity has been typically associated with the processing of pain-related motor responses. However, the potential contribution of putamen to the processing of sensory aspects of pain remains poorly characterized. In order to more directly determine if the putamen can contribute to sensory aspects of pain, nine individuals with lesions involving the putamen underwent both psychophysical and functional imaging assessment of perceived pain and pain-related brain activation. These individuals exhibited intact tactile thresholds, but reduced heat pain sensitivity and widespread reductions in pain-related cortical activity in comparison with 14 age matched healthy subjects. Using magnetic resonance imaging to assess structural connectivity in healthy subjects, we show that portions of the putamen activated during pain are connected not only with cortical regions involved in sensory motor processing, but also regions involved in attention, memory and affect. Such a framework may allow cognitive information to flow from these brain areas to the putamen where it may be used to influence how nociceptive information is processed. Taken together, these findings indicate that the putamen and the basal ganglia may contribute importantly to the shaping of an individual subjective sensory experience by utilizing internal cognitive information to influence activity of large areas of the cerebral cortex. PMID- 21616965 TI - Brain. Editorial. PMID- 21616964 TI - Ovarian metabolism of xenobiotics. AB - At birth, the mammalian ovary contains a finite number of primordial follicles, which once depleted, cannot be replaced. Xenobiotic exposures can destroy primordial follicles resulting in premature ovarian failure and, consequently, early entry into menopause. A number of chemical classes can induce premature ovarian failure, including environmental, chemotherapeutic and industrial exposures. While our knowledge on the mechanistic events that occur in the ovary with chemical exposures is increasing, our understanding of the ovary's capacity to metabolize such compounds is less established. This review will focus on three chemicals for which information on ovarian metabolism is known: trichloroethylene, 7,12-dimethylbenz[a]anthracene and 4-vinylcyclohexene. The current state of understanding of ovarian bioactivation and detoxification processes for each will be described. PMID- 21616966 TI - Therapeutic stem cell plasticity orchestrates tissue plasticity. PMID- 21616967 TI - Inherited peripheral neuropathies: a myriad of genes and complex phenotypes. PMID- 21616968 TI - Beta-amyloid activates PARP causing astrocytic metabolic failure and neuronal death. AB - Alzheimer's disease is characterized by beta-amyloid accumulation in the central nervous system. As beta-amyloid is neurotoxic in culture, we have explored the mechanisms of toxicity in the search for therapeutic targets for Alzheimer's disease and now identify a key role for poly(ADP-ribose) polymerase in beta amyloid-induced neuronal death. Exposure of hippocampal neuronal/glial co cultures to beta-amyloid peptides activates the glial nicotinamide adenine dinucleotide phosphate oxidase, followed by predominantly neuronal cell death. beta-amyloid exposure caused the progressive loss of mitochondrial membrane potential in astrocytes, accompanied by transient mitochondrial depolarizations caused by reversible openings of the mitochondrial permeability transition pore. The transients were absent in cultures from cyclophilin D knockout mice, leaving the slow depolarization available for study in isolation. beta-amyloid exposure decreased both nicotinamide adenine dinucleotide fluorescence and oxygen consumption, while provision of mitochondrial substrates reversed the depolarization, suggesting that substrate supply was limiting. Poly(ADP-ribose) polymerase is activated by oxidative stress and consumes nicotinamide adenine dinucleotide, decreasing substrate availability. beta-amyloid exposure caused accumulation of the poly(ADP-ribose) polymerase product, poly-ADP-ribose polymers, in astrocytes. Inhibition of either poly(ADP-ribose) polymerase or of the nicotinamide adenine dinucleotide phosphate oxidase prevented the appearance of poly-ADP-ribose polymers and the mitochondrial depolarization. Exposure of co cultures to beta-amyloid for >8 h decreased nicotinamide adenine dinucleotide and mitochondrial membrane potential and increased cell death in neurons, all of which were prevented by poly(ADP-ribose) polymerase inhibitors. Poly-ADP-ribose polymers increased with age in the brains of the TASTPM Alzheimer mouse model. We conclude that beta-amyloid-induced neuronal death is mediated by poly(ADP-ribose) polymerase in response to oxidative stress generated by the astrocytic nicotinamide adenine dinucleotide phosphate oxidase. PMID- 21616969 TI - Motor system hyperconnectivity in juvenile myoclonic epilepsy: a cognitive functional magnetic resonance imaging study. AB - Juvenile myoclonic epilepsy is the most frequent idiopathic generalized epilepsy syndrome. It is characterized by predominant myoclonic jerks of upper limbs, often provoked by cognitive activities, and typically responsive to treatment with sodium valproate. Neurophysiological, neuropsychological and imaging studies in juvenile myoclonic epilepsy have consistently pointed towards subtle abnormalities in the medial frontal lobes. Using functional magnetic resonance imaging with an executive frontal lobe paradigm, we investigated cortical activation patterns and interaction between cortical regions in 30 patients with juvenile myoclonic epilepsy and 26 healthy controls. With increasing cognitive demand, patients showed increasing coactivation of the primary motor cortex and supplementary motor area. This effect was stronger in patients still suffering from seizures, and was not seen in healthy controls. Patients with juvenile myoclonic epilepsy showed increased functional connectivity between the motor system and frontoparietal cognitive networks. Furthermore, we found impaired deactivation of the default mode network during cognitive tasks with persistent activation in medial frontal and central regions in patients. Coactivation in the motor cortex and supplementary motor area with increasing cognitive load and increased functional coupling between the motor system and cognitive networks provide an explanation how cognitive effort can cause myoclonic jerks in juvenile myoclonic epilepsy. The supplementary motor area represents the anatomical link between these two functional systems, and our findings may be the functional correlate of previously described structural abnormalities in the medial frontal lobe in juvenile myoclonic epilepsy. PMID- 21616970 TI - Independent oscillatory patterns determine performance fluctuations in children with attention deficit/hyperactivity disorder. AB - The maintenance of stable goal-directed behaviour is a hallmark of conscious executive control in humans. Notably, both correct and error human actions may have a subconscious activation-based determination. One possible source of subconscious interference may be the default mode network that, in contrast to attentional network, manifests intrinsic oscillations at very low (<0.1 Hz) frequencies. In the present study, we analyse the time dynamics of performance accuracy to search for multisecond periodic fluctuations of error occurrence. Attentional lapses in attention deficit/hyperactivity disorder are proposed to originate from interferences from intrinsically oscillating networks. Identifying periodic error fluctuations with a frequency<0.1 Hz in patients with attention deficit/hyperactivity disorder would provide a behavioural evidence for such interferences. Performance was monitored during a visual flanker task in 92 children (7- to 16-year olds), 47 with attention deficit/hyperactivity disorder, combined type and 45 healthy controls. Using an original approach, the time distribution of error occurrence was analysed in the frequency and time-frequency domains in order to detect rhythmic periodicity. Major results demonstrate that in both patients and controls, error behaviour was characterized by multisecond rhythmic fluctuations with a period of ~12 s, appearing with a delay after transition to task. Only in attention deficit/hyperactivity disorder, was there an additional 'pathological' oscillation of error generation, which determined periodic drops of performance accuracy each 20-30 s. Thus, in patients, periodic error fluctuations were modulated by two independent oscillatory patterns. The findings demonstrate that: (i) attentive behaviour of children is determined by multisecond regularities; and (ii) a unique additional periodicity guides performance fluctuations in patients. These observations may re-conceptualize the understanding of attentive behaviour beyond the executive top-down control and may reveal new origins of psychopathological behaviours in attention deficit/hyperactivity disorder. PMID- 21616971 TI - Infarction of 'non-core-non-penumbral' tissue after stroke: multivariate modelling of clinical impact. AB - There is considerable intersubject variability in early neurological course after anterior circulation stroke, yet the pathophysiology underlying this variability is not fully understood. Here, we hypothesize that, although not predicted by current pathophysiological models, infarction of 'non-core-non-penumbral' (i.e. clinically silent) brain tissue may nevertheless occur, and negatively influence clinical course over and above the established positive impact of penumbral salvage. In order to test this hypothesis, non-core-non-penumbral tissue was identified in two independent prospectively recruited cohorts, using computed tomography perfusion, and magnetic resonance perfusion- and diffusion-weighted imaging, respectively. Follow-up structural magnetic resonance imaging was obtained about 1 month later in all patients to map the final infarct. The volumes of both the acutely silent but eventually infarcted tissue, and the eventually non-infarcted penumbra, were determined by performing voxel-wise analysis of the acute and follow-up image sets, using previously validated perfusion thresholds. Early neurological course was expressed as change in National Institutes of Health Stroke Scale scores between the acute and 1-month assessments, relative to the acute score. The relationship between the acutely silent but eventually infarcted tissue volume and early neurological course was tested using a multivariate regression model that included the volume of non infarcted penumbra. Thirty-four and 58 patients were recruited in the computed tomography perfusion and magnetic resonance perfusion cohorts, respectively (mean onset-to-imaging time: 136 and 156 min; 27 and 42 patients received intravenous thrombolysis, respectively). Infarction of acutely silent tissue was identified in most patients in both cohorts. Although its volume (median 0.2 and 2 ml, respectively) was much smaller than that of salvaged penumbra (59.3 and 93 ml, respectively), it was substantial in ~10% of patients. As expected, salvaged penumbra strongly positively influenced early neurological course. Even after correcting for the latter effect in the multivariate model, infarction of acutely silent tissue independently negatively influenced early neurological course in both cohorts (P=0.018 and 0.031, respectively). This is the first systematic study to document infarction of acutely silent tissue after anterior circulation stroke, and to show that it affects a sizeable fraction of patients and has the predicted negative impact on clinical course. These findings were replicated in two independent cohorts, regardless of the perfusion imaging modality used. Preventing infarction of the tissue not initially at risk should have direct clinical benefit. PMID- 21616973 TI - Inherited prion disease with 4-octapeptide repeat insertion: disease requires the interaction of multiple genetic risk factors. AB - Genetic factors are implicated in the aetiology of sporadic late-onset neurodegenerative diseases. Whether these genetic variants are predominantly common or rare, and how multiple genetic factors interact with each other to cause disease is poorly understood. Inherited prion diseases are highly heterogeneous and may be clinically mistaken for sporadic Creutzfeldt-Jakob disease because of a negative family history. Here we report our investigation of patients from the UK with four extra octapeptide repeats, which suggest that the risk of clinical disease is increased by a combination of the mutation and a susceptibility haplotype on the wild-type chromosome. The predominant clinical syndrome is a progressive cortical dementia with pyramidal signs, myoclonus and cerebellar abnormalities that closely resemble sporadic Creutzfeldt-Jakob disease. Autopsy shows perpendicular deposits of prion protein in the molecular layer of the cerebellum. Identity testing, PRNP microsatellite haplotyping and genealogical work confirm no cryptic close family relationships and suggests multiple progenitor disease haplotypes. All patients were homozygous for methionine at polymorphic codon 129. In addition, at a single nucleotide polymorphism upstream of PRNP thought to confer susceptibility to sporadic Creutzfeldt-Jakob disease (rs1029273), all patients were homozygous for the risk allele (combined P=5.9*10(-5)). The haplotype identified may also be a risk factor in other partially penetrant inherited prion diseases although it does not modify age of onset. Blood expression of PRNP in healthy individuals was modestly higher in carriers of the risk haplotype. These findings may provide a precedent for understanding apparently sporadic neurodegenerative diseases caused by rare high-risk mutations. PMID- 21616972 TI - Human neural stem cells enhance structural plasticity and axonal transport in the ischaemic brain. AB - Stem cell transplantation promises new hope for the treatment of stroke although significant questions remain about how the grafted cells elicit their effects. One hypothesis is that transplanted stem cells enhance endogenous repair mechanisms activated after cerebral ischaemia. Recognizing that bilateral reorganization of surviving circuits is associated with recovery after stroke, we investigated the ability of transplanted human neural progenitor cells to enhance this structural plasticity. Our results show the first evidence that human neural progenitor cell treatment can significantly increase dendritic plasticity in both the ipsi- and contralesional cortex and this coincides with stem cell-induced functional recovery. Moreover, stem cell-grafted rats demonstrated increased corticocortical, corticostriatal, corticothalamic and corticospinal axonal rewiring from the contralesional side; with the transcallosal and corticospinal axonal sprouting correlating with functional recovery. Furthermore, we demonstrate that axonal transport, which is critical for both proper axonal function and axonal sprouting, is inhibited by stroke and that this is rescued by the stem cell treatment, thus identifying another novel potential mechanism of action of transplanted cells. Finally, we established in vitro co-culture assays in which these stem cells mimicked the effects observed in vivo. Through immunodepletion studies, we identified vascular endothelial growth factor, thrombospondins 1 and 2, and slit as mediators partially responsible for stem cell-induced effects on dendritic sprouting, axonal plasticity and axonal transport in vitro. Thus, we postulate that human neural progenitor cells aid recovery after stroke through secretion of factors that enhance brain repair and plasticity. PMID- 21616975 TI - Effect of adjunctive treatment with aripiprazole to atypical antipsychotics on cognitive function in schizophrenia patients. AB - Second-generation antipsychotics yield only a modest improvement in cognitive benefit compared to first-generation antipsychotics. Aripiprazole, which is a partial dopamine D2 receptor agonist, may have an impact on cognitive dysfunction in patients with schizophrenia. This study administered aripiprazole or placebo to 36 outpatients with schizophrenia also receiving risperidone or olanzapine for 12 weeks in a double-blind, randomized, placebo-controlled study. Cognitive function was evaluated using the Brief Assessment of Cognition in Schizophrenia (BACS) just prior to drug administration as well as 12 weeks after. The PANSS and UKU side effect rating scales were used to evaluate the clinical response to additional treatment with aripiprazole. In a primary analyses, ANCOVA showed that there was an interaction between the treatment group and time for verbal fluency (p < 0.05), but not for any domain in BACS, PANSS or UKU side effect rating scales. Upon secondary analysis, however, the ameliorative change in motor speed as assessed by the BACS (p < 0.05) for those receiving aripiprazole was greater than that for the placebo group, whereas deterioration in verbal fluency (p < 0.01) and executive function (p < 0.01) in those receiving aripiprazole was significantly greater than in the placebo group. These results suggest that adjunctive treatment with aripiprazole improves motor speed but worsens some cognitive functions. It is likely that these effects are due to the dopamine D2 antagonistic effect of aripiprazole. PMID- 21616976 TI - Neuroanatomical substrates involved in cannabinoid modulation of defensive responses. AB - Administration of Cannabis sativa derivatives causes anxiolytic or anxiogenic effects in humans and laboratory animals, depending on the specific compound and dosage used. In agreement with these findings, several studies in the last decade have indicated that the endocannabinoid system modulates neuronal activity in areas involved in defensive responses. The mechanisms of these effects, however, are still not clear. The present review summarizes recent data suggesting that they involve modulation of glutamate and GABA-mediated neurotransmission in brain sites such as the medial prefrontal cortex, amygdaloid complex, bed nucleus of the stria terminalis, hippocampus and dorsal periaqueductal gray. Moreover, we also discuss results indicating that, in these regions, the endocannabinoid system could be particularly engaged by highly stressful situations. PMID- 21616974 TI - Impaired neurovascular repair in subjects with diabetes following experimental intracutaneous axotomy. AB - Diabetic complications and vascular disease are closely intertwined. Diabetes mellitus is a well-established risk factor for both large and small vessel vascular changes, and conversely other vascular risk factors confer increased risk for diabetic complications such as peripheral neuropathy, nephropathy and retinopathy. Furthermore, axons and blood vessels share molecular signals for purposes of navigation, regeneration and terminal arborizations. We examined blood vessel, Schwann cell and axonal regeneration using validated axotomy models to study and compare patterns and the relationship of regeneration among these different structures. Ten subjects with diabetes mellitus complicated by neuropathy and 10 healthy controls underwent 3 mm distal thigh punch skin biopsies to create an intracutaneous excision axotomy followed by a concentric 4 mm overlapping biopsy at different time points. Serial sections were immunostained against a pan-axonal marker (PGP9.5), an axonal regenerative marker (GAP43), Schwann cells (p75) and blood vessels (CD31) to visualize regenerating structures in the dermis and epidermis. The regenerative and collateral axonal sprouting rates, blood vessel growth rate and Schwann cell density were quantified using established stereology techniques. Subjects also underwent a chemical 'axotomy' through the topical application of capsaicin, and regenerative sprouting was assessed by the return of intraepidermal nerve fibre density through regenerative regrowth. In the healed 3 mm biopsy sites, collateral and dermal regenerative axonal sprouts grew into the central denervated area in a stereotypic pattern with collateral sprouts growing along the dermal-epidermal junction while regenerative dermal axons, blood vessels and Schwann cells grew from their transected proximal stumps into the deep dermis. Vessel growth preceded axon and Schwann cell migration into the denervated region, perhaps acting as scaffolding for axon and Schwann cell growth. In control subjects, Schwann cell growth was more robust and extended into the superficial dermis, while among subjects with diabetes mellitus, Schwann tubes appeared atrophic and were limited to the mid-dermis. Rates of collateral (P=0.0001), dermal axonal regenerative sprouting (P=0.02), Schwann cell migration (P<0.05) and blood vessel growth (P=0.002) were slower among subjects with diabetes mellitus compared with control subjects. Regenerative deficits are a common theme in diabetes mellitus and may underlie the development of neuropathy. We observed that blood vessel growth recapitulated the pattern seen in ontogeny and preceded regenerating nerve fibres, suggesting that enhancement of blood vessel growth might facilitate axonal regeneration. These models are useful tools for the efficient investigation of neurotrophic and regenerative drugs, and also to explore factors that may differentially affect axonal regeneration. PMID- 21616977 TI - MDMA intoxication and verbal memory performance: a placebo-controlled pharmaco MRI study. AB - The aim of the present study was to identify the neural substrate underlying memory impairment due to a single dose of MDMA (3,4 methylenedioxymethamphetamine) by means of pharmaco-MRI. Based on previous behavioral results it was hypothesized that this deficit could be attributed to a specific influence of MDMA on encoding. Fourteen Ecstasy users participated in this double-blind, placebo-controlled, within-subject study with two treatment conditions: MDMA (75 mg) and placebo. Memory performance was tested by means of a word learning task including two words lists, one addressing reading processes (control task, CWL) and a second (experimental task, EWL) addressing encoding and reading processes. Behavioral data showed that under the influence of MDMA, EWL performance was worse than placebo. Imaging data showed that Encoding was situated mainly in (pre)frontal, temporal and parietal areas. MDMA by Encoding interaction was situated in three areas: the left middle frontal gyrus (BA10), the right fusiform gyrus (BA19), and the left cuneus (BA18). Behavioral and functional data only correlated in BA10. It appeared that EWL performance caused BOLD signal change in BA10 during placebo treatment but not during MDMA intoxication. It is concluded that MDMA influences middle frontal gyrus processes resulting in impoverished memory encoding. PMID- 21616978 TI - Nicotine restores morphine-induced memory deficit through the D1 and D2 dopamine receptor mechanisms in the nucleus accumbens. AB - Involvement of the dopamine D1 and D2 receptors in the nucleus accumbens (NAc) with interaction between morphine and nicotine on inhibitory avoidance (IA) memory was investigated. A step-through type of inhibitory avoidance tasks was used to assess memory in male Wistar rats. The results showed that subcutaneous (s.c.) administration of morphine (7.5 mg/kg) after training decreased retrieval of IA memory in the animals when tested 24 h later. Pre-test administration of the same dose of morphine significantly reversed the deficiency in retrieval. The results also showed that pre-test administration of nicotine (0.2 and 0.4 mg/kg, s.c.) by itself mimicked the effect of pre-test morphine, and lower doses of nicotine (0.1 and 0.2 mg/kg) also improved the effect of a low dose of morphine (2.5 mg/kg) on retrieval of IA memory. Pre-test intra-NAc administration of the dopamine D1 receptor antagonist, SCH 23390 (0.001 and 0.01 ug/rat), and the dopamine D2 receptor antagonist, sulpiride (0.5 and 1 ug/rat) caused no significant effects on IA memory by themselves, but both prevented reinstatement of the retrieval of IA memory by the effective dose of nicotine (0.4 mg/kg). It can be concluded that the dopaminergic mechanism(s) in the NAc is a crosslink for the effect of morphine and nicotine on reinstatement of retrieval of IA memory impaired by post-training administration of morphine. PMID- 21616979 TI - Lack of effect of ketamine on cortical glutamate and glutamine in healthy volunteers: a proton magnetic resonance spectroscopy study. AB - Ketamine is a N-methyl-D-aspartic acid (NMDA) antagonist that has been associated with temporary clinical improvement in patients with depression. Studies using magnetic resonance spectroscopy (MRS) have shown that major depression is associated with decreased levels of glutamate and glutamine (Glx) in the anterior cingulate cortex, which normalize with clinical recovery. The present study aimed to test whether a ketamine infusion would increase cortical Glx levels in healthy volunteers. Healthy volunteers received an intravenous infusion of ketamine (0.5 mg kg-1, n = 8) or saline (n = 9) over 40 minutes. MRS measurements were obtained at baseline, during, and at the end of the infusion. The infusion of ketamine had significant effects on mental state but there was no effect of ketamine on the levels of Glx (F (3,39) = 1.70, p = 0.18) or glutamate (F (3,39) = 48, p = 0.70). This study suggests that the gradual infusion of low-dose ketamine in antidepressant doses not cause changes in cortical glutamate or glutamine in healthy volunteers that are visible by proton MRS. PMID- 21616981 TI - Prediction of the outcome of orthodontic treatment of Class III malocclusion. PMID- 21616982 TI - Decoding subject-driven cognitive states with whole-brain connectivity patterns. AB - Decoding specific cognitive states from brain activity constitutes a major goal of neuroscience. Previous studies of brain-state classification have focused largely on decoding brief, discrete events and have required the timing of these events to be known. To date, methods for decoding more continuous and purely subject-driven cognitive states have not been available. Here, we demonstrate that free-streaming subject-driven cognitive states can be decoded using a novel whole-brain functional connectivity analysis. Ninety functional regions of interest (ROIs) were defined across 14 large-scale resting-state brain networks to generate a 3960 cell matrix reflecting whole-brain connectivity. We trained a classifier to identify specific patterns of whole-brain connectivity as subjects rested quietly, remembered the events of their day, subtracted numbers, or (silently) sang lyrics. In a leave-one-out cross-validation, the classifier identified these 4 cognitive states with 84% accuracy. More critically, the classifier achieved 85% accuracy when identifying these states in a second, independent cohort of subjects. Classification accuracy remained high with imaging runs as short as 30-60 s. At all temporal intervals assessed, the 90 functionally defined ROIs outperformed a set of 112 commonly used structural ROIs in classifying cognitive states. This approach should enable decoding a myriad of subject-driven cognitive states from brief imaging data samples. PMID- 21616983 TI - New pool of cortical interneuron precursors in the early postnatal dorsal white matter. AB - The migration of cortical gamma-aminobutyric acidergic interneurons has been extensively studied in rodent embryos, whereas few studies have documented their postnatal migration. Combining in vivo analysis together with time-lapse imaging on cortical slices, we explored the origin and migration of cortical interneurons during the first weeks of postnatal life. Strikingly, we observed that a large pool of GAD65-GFP-positive cells accumulate in the dorsal white matter region during the first postnatal week. Part of these cells divides and expresses the transcription factor paired box 6 indicating the presence of local transient amplifying precursors. The vast majority of these cells are immature interneurons expressing the neuronal marker doublecortin and partly the calcium-binding protein calretinin. Time-lapse imaging reveals that GAD65-GFP-positive neurons migrate from the white matter pool into the overlying anterior cingulate cortex (aCC). Some interneurons in the postnatal aCC express the same immature neuronal markers suggesting ongoing migration of calretinin-positive interneurons. Finally, bromodeoxyuridine incorporation experiments confirm that a small fraction of interneurons located in the aCC are generated during the early postnatal period. These results altogether reveal that at postnatal ages, the dorsal white matter contains a pool of interneuron precursors that divide and migrate into the aCC. PMID- 21616985 TI - The contribution of emotion and cognition to moral sensitivity: a neurodevelopmental study. AB - Whether emotion is a source of moral judgments remains controversial. This study combined neurophysiological measures, including functional magnetic resonance imaging, eye-tracking, and pupillary response with behavioral measures assessing affective and moral judgments across age. One hundred and twenty-six participants aged between 4 and 37 years viewed scenarios depicting intentional versus accidental actions that caused harm/damage to people and objects. Morally, salient scenarios evoked stronger empathic sadness in young participants and were associated with enhanced activity in the amygdala, insula, and temporal poles. While intentional harm was evaluated as equally wrong across all participants, ratings of deserved punishments and malevolent intent gradually became more differentiated with age. Furthermore, age-related increase in activity was detected in the ventromedial prefrontal cortex in response to intentional harm to people, as well as increased functional connectivity between this region and the amygdala. Our study provides evidence that moral reasoning involves a complex integration between affective and cognitive processes that gradually changes with age and can be viewed in dynamic transaction across the course of ontogenesis. The findings support the view that negative emotion alerts the individual to the moral salience of a situation by bringing discomfort and thus can serve as an antecedent to moral judgment. PMID- 21616984 TI - The effects of aging on material-independent and material-dependent neural correlates of source memory retrieval. AB - Age-related declines in source memory have been observed for various stimuli and associated details. These impairments may be related to alterations in brain regions contributing to source memory via material-independent processes and/or regions specialized for processing specific materials. Using event-related functional magnetic resonance imaging, we investigate the effects of aging on source memory and associated neural activity for words and objects. Source accuracy was equally impaired in older adults for both materials. Imaging data revealed both groups recruited similar networks of regions to support source memory accuracy irrespective of material, including parietal and prefrontal cortices (PFC) and the hippocampus. Age-related decreases in material-independent activity linked to postretrieval monitoring were observed in right lateral PFC. Additionally, age-related increases in source accuracy effects were shown in perirhinal cortex, which were positively correlated with performance in older adults, potentially reflecting functional compensation. In addition to group differences in material-independent regions, age-related crossover interactions for material-dependent source memory effects were observed in regions selectively engaged by objects. These results suggest that older adults' source memory impairments reflect alterations in regions making material-independent contributions to source memory retrieval, primarily the lateral PFC, but may be further impacted by changes in regions sensitive to particular materials. PMID- 21616986 TI - Program intensity and service delivery models in the schools: SLP survey results. AB - PURPOSE: School-based speech-language pathologists (SLPs) routinely work with team members to make recommendations regarding an intervention program's intensity and method of service delivery for children with speech and language impairments. In this study, student, SLP, and workplace characteristics that may influence SLPs' recommendations were examined. METHOD: Almost 2,000 school-based SLPs completed an online survey about the factors they consider when making recommendations regarding program intensity and service delivery model that students on their caseloads receive. RESULTS: SLPs reported that student characteristics, rather than SLP or workplace characteristics, were the factors they considered the most when making these recommendations. However, these same SLPs reported that current students on their caseload with severe to moderate disabilities participated in intervention 2-3 times a week for 20-30 min in groups outside of the classroom. Students with the least severe disability received intervention 1 time a week for 20-30 min in groups outside of the classroom. CONCLUSION: The limited variety of intervention program intensities and service delivery models used suggests that student characteristics may not be the most important factor considered when making intervention recommendations, as reported by the SLPs. Instead, caseload size and years of practice appear to influence SLPs' recommendations regarding which program intensity and service delivery models to use. PMID- 21616987 TI - Addressing clinician-client mismatch: a preliminary intervention study with a bilingual Vietnamese-English preschooler. AB - PURPOSE: This project examined receptive vocabulary treatment outcomes in the two languages of a bilingual preschooler with moderate to severe language impairment. METHOD: A series of single-subject experimental designs was used to compare English-only (EO) and bilingual (BI) approaches to receptive vocabulary treatment. The participant, Nam, was a boy age 3;11 (years;months) who was learning Vietnamese as a first language at home and English in his early childhood education program. Treatment was implemented by an EO interventionist using a computer interface and prerecorded audio files in Vietnamese and English. The dependent measure was the percentage of items that were correctly identified in each language. RESULTS: Combined studies revealed that the BI approach increased Nam's attention to task and was as effective as the EO approach for increasing his receptive vocabulary in English. Nam made vocabulary gains in both treatment conditions; receptive vocabulary gains were evident in both Vietnamese and English. CONCLUSION: This project showed that it is feasible for an EO clinician to promote gains in both the home and school languages of a BI child through creative collaborations with BI colleagues and the use of technology. Replication with additional participants and treatment activities is needed to make further generalizations. PMID- 21616988 TI - Increasing the odds: applying emergentist theory in language intervention. AB - PURPOSE: This review introduces emergentism, which is a leading theory of language development that states that language ability is the product of interactions between the child's language environment and his or her learning capabilities. The review suggests ways in which emergentism provides a theoretical rationale for interventions that are designed to address developmental language delays in young children. METHOD: A review of selected literature on emergentist theory and research is presented, with a focus on the acquisition of early morphology and syntax. A significant method for developing and testing emergentist theory, connectionist modeling, is described. Key themes from both connectionist and behavioral studies are summarized and applied with specific examples to language intervention techniques. A case study is presented to integrate elements of emergentism with language intervention. CONCLUSIONS: Evaluating the theoretical foundation for language interventions is an important step in evidence-based practice. This article introduces three themes in the emergentist literature that have implications for language intervention: (a) sufficiency of language input, (b) active engagement of the child with the input, and (c) factors that increase the odds for correctly mapping language form to meaning. Evidence supporting the importance of these factors in effective language intervention is presented, along with limitations in that evidence. PMID- 21616989 TI - The use of sound-field amplification devices in different types of classrooms. AB - PURPOSE: The purpose of this study was to determine if sound-field amplification (SFA) devices affected student performance in 3 different types of classrooms. METHOD: The classroom performance of 147 children (77 males, 70 females, ages 8;2 [years;months] +/- 5 months) was measured at the beginning and end of the second semester of their third year in 1 of 4 primary schools in Brisbane, Australia. Each school contained 2 participating classrooms, 1 with and 1 without an SFA device. RESULTS: The SFA devices contributed to small but significant improvements in student listening (p < .01) and auditory analysis (p < .05) skills, but only in the school where the participating classrooms were in a brick building (vs. a demountable building) with neighboring classrooms separated by solid walls (vs. open spaces). The classrooms in this school showed the lowest background noise measures (47-50 dB 1 hr, A weighted) and the second lowest reverberation times (0.87-0.91 s) overall, although these values still exceeded the maximums recommended by American National Standards Institute S12.60-2002 (2002). CONCLUSION: These results suggest that any potential benefits of SFA devices are more likely to be realized in classrooms with better acoustics. PMID- 21616990 TI - Change in marital status structure as an obstacle for health improvement: evidence from six developed countries. AB - The aim of this study is to estimate the contributions of changes in population distribution by marital status to the changes in adult mortality in six developed countries. The change in total mortality was decomposed into the contributions of: (i) mortality changes within each marital status category; and (ii) changes in population composition by marital status. The study provides evidence that changes in population composition contributed to increases in male mortality in Russia and Lithuania, whereas in the remaining male populations this was a significant obstacle for faster health improvements. The compositional changes had only small impacts on female mortality. PMID- 21616991 TI - Socio-economic disadvantage and body mass over the life course in women and men: results from the Northern Swedish Cohort. AB - BACKGROUND: Obesity and body mass in adulthood relate both to current and to childhood socio-economic status, particularly in women, but the underlying life course processes are not known. This study aims at examining whether the life course socio-economic status-body mass association in women and men is explained by the cumulative risk or adolescent sensitive period models whether associations are similar at different life course stages; and whether health behaviours explain the associations. METHODS: A total of 476 women and 517 men participated in this 27-year prospective cohort study (participation rate 93%). Body mass index was assessed at the age of 16 and 43 years and self-reported at the age of 21 and 30 years. Information on socio-economic status by own or parental (age 16 years) occupation, smoking, snuff, alcohol, physical activity and diet was collected at each age. RESULTS: In women, cumulative socio-economic status and socio-economic status in adolescence were related to body mass index at the age of 16, 21, 30 and 43 years and to the 27-year change in body mass, independently of health behaviours and for adolescent socio-economic status also of later socio economic attainment. Associations were generally stronger for body mass at older age. In men, associations were mostly non-significant, although health behaviours contributed strongly to body mass. CONCLUSIONS: In women, both the sensitive period (in adolescence) and cumulative risk models explain the socio-economic body mass link. Efforts to reduce the social inequality in body mass in women should be directed at the early life course, but focusing on unhealthy behaviours might not be a sufficient approach. PMID- 21616992 TI - Breastfeeding and obesity in Brazilian children. AB - BACKGROUND: The association between breastfeeding and obesity is inconsistent by the literature. This study aims to assess whether obesity is associated to occurrence of breastfeeding and to duration of total and exclusive breastfeeding in Brazilian children. METHODS: A cross-sectional study was conducted with 764 children enrolled in public and private schools from Vicosa, Minas Gerais, Brazil. Obesity (outcome variable) was defined as body mass index above the +2 standard deviations score using sex and age specific standards of World Health Organization. Exposure was the occurrence and duration of breastfeeding. Potential confounders were controlled by multiple logistic regression analysis and were divided in two groups: children (gender, age, birth weight, gestational age, order of birth, number of siblings, number of persons in the residence, type of school, physical activity patterns and time watching television) and mothers (age, nutritional status, level of education, weight gain during pregnancy, smokes currently and during the pregnancy). RESULTS: Prevalence of obesity was 10.7%; 6.8% of the children were not breastfed and 59.0% did not receive exclusive breastfeeding. After adjustment for confounding variables by logistic regression analysis, no statistically significant association was observed between obesity and the occurrence and/or duration of total and exclusive breastfeeding. There was no dose-response effect of duration of breastfeeding on prevalence of obesity. CONCLUSION: Our results do not support the hypothesis that breastfeeding promotion would reduce obesity in this population. Controversial findings regarding this association by literature indicate a need for further investigations. PMID- 21616993 TI - Tobacco consumption and pancreatic cancer mortality: what can we conclude from historical data in Australia? AB - BACKGROUND: Tobacco consumption is an established risk factor for pancreatic cancer yet studies of long-term mortality trends have not statistically analysed this relationship. We sought evidence for this relationship based on an analysis of long-term population-level data in Australia. METHODS: Pancreatic cancer mortality data from 1931, tobacco consumption data and fruit and vegetable consumption data for Australia were utilized. Log-linear Poisson regression models were used to analyse pancreatic cancer mortality from 1931 with cumulative cohort and lagged time-specific tobacco consumption data and fruit and vegetable consumption data. RESULTS: Pancreatic cancer mortality rose steadily for males until it began falling from the 1970s, and continued rising for females until 2006. These trends correspond with a long-term rise in male tobacco consumption until the 1960s and a later peak for females. Our models show that cumulative tobacco consumption predicts pancreatic cancer mortality for both sexes but with time lags only being significant for males. Fruit and vegetable consumption provides a protective effect against mortality in some of the models. CONCLUSION: The success of smoking reduction programmes in Australia has contributed to the decline in pancreatic cancer mortality for males, providing important evidence about the need for tobacco control measures in populations where it is still increasing. Continued declines in female tobacco consumption should lead to a reversal of the long-term rise in female pancreatic cancer mortality. PMID- 21616994 TI - Elevated plasma growth differentiation factor-15 correlates with lymph node metastases and poor survival in endometrial cancer. AB - PURPOSE: The study objective was to investigate and validate plasma growth differentiation factor-15 (GDF-15) as a predictor of lymph node metastasis and a poor prognosis in primary endometrial cancer. EXPERIMENTAL DESIGN: Plasma samples from 510 women treated for endometrial cancer in a primary investigation cohort (n = 44) and a secondary validation cohort (n = 466) were analyzed for GDF-15. Plasma from healthy premenopausal (n = 20) and postmenopausal (n = 20) women, women with borderline (n = 43), benign (n = 144), and malignant ovarian tumors (n = 125) were used for comparison. RESULTS: Median plasma GDF-15 concentration for the endometrial cancer group was elevated (1,077 ng/L) as compared with pre- and postmenopausal controls (590 and 684 ng/L) and women with benign (591 ng/L) or borderline ovarian tumors (718 ng/L; all P < 0.001), but similar to the ovarian cancer group. In the large validation cohort of endometrial carcinomas, high plasma GDF-15 was significantly associated with International Federation of Gynecology and Obstetrics (FIGO) stage III/IV disease, nonendometrioid histology, high grade, older age, postmenopausal status, and lymph node metastases (all P <= 0.001). High GDF-15 was also an independent predictor of poor disease-specific and recurrence-free survival. CONCLUSIONS: Based on findings indicated in a primary investigation set and confirmed in the large secondary validation set, we report for the first time plasma GDF-15 as a biomarker for endometrial cancer phenotype, including presence of lymph node metastasis and reduced survival. Its applicability as a predictor of metastatic nodes and in monitoring treatment of endometrial cancer needs to be further studied. PMID- 21616995 TI - Peripheral vasoconstriction and abnormal parasympathetic response to sighs and transient hypoxia in sickle cell disease. AB - RATIONALE: Sickle cell disease is an inherited blood disorder characterized by vasoocclusive crises. Although hypoxia and pulmonary disease are known risk factors for these crises, the mechanisms that initiate vasoocclusive events are not well known. OBJECTIVES: To study the relationship between transient hypoxia, respiration, and microvascular blood flow in patients with sickle cell. METHODS: We established a protocol that mimics nighttime hypoxic episodes and measured microvascular blood flow to determine if transient hypoxia causes a decrease in microvascular blood flow. Significant desaturations were induced safely by five breaths of 100% nitrogen. MEASUREMENTS AND MAIN RESULTS: Desaturation did not induce change in microvascular perfusion; however, it induced substantial transient parasympathetic activity withdrawal in patients with sickle cell disease, but not controls subjects. Marked periodic drops in peripheral microvascular perfusion, unrelated to hypoxia, were triggered by sighs in 11 of 11 patients with sickle cell and 8 of 11 control subjects. Although the sigh frequency was the same in both groups, the probability of a sigh inducing a perfusion drop was 78% in patients with sickle cell and 17% in control subjects (P < 0.001). Evidence for sigh-induced sympathetic nervous system dominance was seen in patients with sickle cell (P < 0.05), but was not significant in control subjects. CONCLUSIONS: These data demonstrate significant disruption of autonomic nervous system balance, with marked parasympathetic withdrawal in response to transient hypoxia. They draw attention to an enhanced autonomic nervous system mediated sigh-vasoconstrictor response in patients with sickle cell that could increase red cell retention in the microvasculature, promoting vasoocclusion. PMID- 21616996 TI - Prompting physicians to address a daily checklist and process of care and clinical outcomes: a single-site study. AB - RATIONALE: Checklists may reduce errors of omission for critically ill patients. OBJECTIVES: To determine whether prompting to use a checklist improves process of care and clinical outcomes. METHODS: We conducted a cohort study in the medical intensive care unit (MICU) of a tertiary care university hospital. Patients admitted to either of two independent MICU teams were included. Intervention team physicians were prompted to address six parameters from a daily rounding checklist if overlooked during morning work rounds. The second team (control) used the identical checklist without prompting. MEASUREMENTS AND MAIN RESULTS: One hundred and forty prompted group patients were compared with 125 control and 1,283 preintervention patients. Compared with control, prompting increased median ventilator-free duration, decreased empirical antibiotic and central venous catheter duration, and increased rates of deep vein thrombosis and stress ulcer prophylaxis. Prompted group patients had lower risk-adjusted ICU mortality compared with the control group (odds ratio, 0.36; 95% confidence interval, 0.13 0.96; P = 0.041) and lower hospital mortality compared with the control group (10.0 vs. 20.8%; P = 0.014), which remained significant after risk adjustment (odds ratio, 0.34; 95% confidence interval, 0.15-0.76; P = 0.008). Observed-to predicted ICU length of stay was lower in the prompted group compared with control (0.59 vs. 0.87; P = 0.02). Checklist availability alone did not improve mortality or length of stay compared with preintervention patients. CONCLUSIONS: In this single-site, preliminary study, checklist-based prompting improved multiple processes of care, and may have improved mortality and length of stay, compared with a stand-alone checklist. The manner in which checklists are implemented is of great consequence in the care of critically ill patients. PMID- 21616999 TI - Ascertainment of individual risk of mortality for patients with idiopathic pulmonary fibrosis. AB - RATIONALE: Several predictors of mortality in patients with idiopathic pulmonary fibrosis have been described; however, there is a need for a practical and accurate method of quantifying the prognosis of individual patients. OBJECTIVES: Develop a practical mortality risk scoring system for patients with idiopathic pulmonary fibrosis. METHODS: We used a Cox proportional hazards model and data from two clinical trials (n = 1,099) to identify independent predictors of 1-year mortality among patients with idiopathic pulmonary fibrosis. From the comprehensive model, an abbreviated clinical model comprised of only those predictors that are readily and reliably ascertained by clinicians was derived. Beta coefficients for each predictor were then used to develop a practical mortality risk scoring system. MEASUREMENTS AND MAIN RESULTS: Independent predictors of mortality included age, respiratory hospitalization, percent predicted FVC, 24-week change in FVC, percent predicted carbon monoxide diffusing capacity, 24-week change in percent predicted carbon monoxide diffusing capacity, and 24-week change in health-related quality of life. An abbreviated clinical model comprising only four predictors (age, respiratory hospitalization, percent predicted FVC, and 24-wk change in FVC), and the corresponding risk scoring system produced estimates of 1-year mortality risk consistent with observed data (9.9% vs. 9.7%; C statistic = 0.75; 95% confidence interval, 0.71-0.79). CONCLUSIONS: The prognosis for patients with idiopathic pulmonary fibrosis may be accurately determined using four readily ascertainable predictors. Our simplified scoring system may be a valuable tool for determining prognosis and guiding clinical management. Additional research is needed to validate the applicability and accuracy of the scoring system. PMID- 21617000 TI - Psychosocial processes and sexual initiation among Ghanaian youth. AB - This study models primary abstinence and age at first sex in a cross-sectional sample of Ghanaian youth aged 17 to 22 years. The aim is to examine how reproductive knowledge and social cognitive factors jointly affect the choice to initiate sex. Among males, the authors find that reproductive knowledge is negatively associated with abstinence. Its negative association, however, is moderated by the extent of adult support a respondent receives. Among females, reproductive knowledge is positively associated with abstinence. Its positive association is moderated by the extent of household sex communication. Correspondingly, when age at first sex is modeled, knowledge is a negative predictor among females. Its effects are mediated by household sex communication. A third social cognitive variable, perceived permissive peer attitudes about sex, is not associated with knowledge but is a protective factor for sexual initiation for both sexes. Though causal direction cannot be established, the results suggest that cognition affects sexual initiation both directly and through complex social mechanisms. The evidence provides theoretical support for comprehensive adolescent sexual and reproductive health interventions that promote abstinence and other safe sex behaviors through social as well as individual pathways. PMID- 21616998 TI - Early alveolar epithelial dysfunction promotes lung inflammation in a mouse model of Hermansky-Pudlak syndrome. AB - RATIONALE: The pulmonary phenotype of Hermansky-Pudlak syndrome (HPS) in adults includes foamy alveolar type 2 cells, inflammation, and lung remodeling, but there is no information about ontogeny or early disease mediators. OBJECTIVES: To establish the ontogeny of HPS lung disease in an animal model, examine disease mediators, and relate them to patients with HPS1. METHODS: Mice with mutations in both HPS1/pale ear and HPS2/AP3B1/pearl (EPPE mice) were studied longitudinally. Total lung homogenate, lung tissue sections, and bronchoalveolar lavage (BAL) were examined for phospholipid, collagen, histology, cell counts, chemokines, surfactant protein D (SP-D), and S-nitrosylated SP-D. Isolated alveolar epithelial cells were examined for expression of inflammatory mediators, and chemotaxis assays were used to assess their importance. Pulmonary function test results and BAL from patients with HPS1 and normal volunteers were examined for clinical correlation. MEASUREMENTS AND MAIN RESULTS: EPPE mice develop increased total lung phospholipid, followed by a macrophage-predominant pulmonary inflammation, and lung remodeling including fibrosis. BAL fluid from EPPE animals exhibited early accumulation of both SP-D and S-nitrosylated SP-D. BAL fluid from patients with HPS1 exhibited similar changes in SP-D that correlated inversely with pulmonary function. Alveolar epithelial cells demonstrated expression of both monocyte chemotactic protein (MCP)-1 and inducible nitric oxide synthase in juvenile EPPE mice. Last, BAL from EPPE mice and patients with HPS1 enhanced migration of RAW267.4 cells, which was attenuated by immunodepletion of SP-D and MCP-1. CONCLUSIONS: Inflammation is initiated from the abnormal alveolar epithelial cells in HPS, and S-nitrosylated SP-D plays a significant role in amplifying pulmonary inflammation. PMID- 21617001 TI - Cellulitis, headache, and fever following tick bites. PMID- 21617002 TI - Targeting the tcdA gene: is this appropriate for detection of A and/or B Clostridium difficile toxin-producing strains? PMID- 21617004 TI - Soul mate: exploring the concept of soul. AB - This article describes an "advanced practice" registered nurse's skill in using multiple theoretical frameworks to make meaning of her severely developmentally disabled son's untimely death. Aspects of religion, spirituality, and philosophy are presented plus how related practices, such as used within Alcoholics Anonymous, are incorporated into everyday life are referenced. Creating unique rituals and ceremonies demonstrates the power of the mind as a partner in the healing process when grief seems insurmountable. This article, titled "Soul Mate" discusses how individuals create their own healing narratives when confronted with grief and tragedy. Nursing interventions, sensitive to this process, support and promote the grief process. Eliciting, recognizing, and accepting a patient's unique self-made rituals and ceremonies as they cope with a beloved's death and dying enhances their nursing interventions. PMID- 21617005 TI - The stingy bone marrow and the wasteful peripheral blood: a tale of two ITPs. PMID- 21617006 TI - Correcting CGD safely, iPSo facto. PMID- 21617007 TI - Hematopoiesis and bone remodeling. PMID- 21617008 TI - Vav-Vav-Vav-voom! PMID- 21616997 TI - Characteristics and outcomes of ventilated patients according to time to liberation from mechanical ventilation. AB - RATIONALE: A new classification of patients based on the duration of liberation of mechanical ventilation has been proposed. OBJECTIVES: To analyze outcomes based on the new weaning classification in a cohort of mechanically ventilated patients. METHODS: Secondary analysis included 2,714 patients who were weaned and underwent scheduled extubation from a cohort of 4,968 adult patients mechanically ventilated for more than 12 hours. MEASUREMENTS AND MAIN RESULTS: Patients were classified according to a new weaning classification: 1,502 patients (55%) as simple weaning,1,058 patients (39%) as difficult weaning, and 154 (6%) as prolonged weaning.Variables associated with prolonged weaning(.7d)were: severity at admission (odds ratio [OR] per unit of Simplified Acute Physiology Score II, 1.01; 95% confidence interval [CI], 1.001-1.02), duration of mechanical ventilation before first attempt of weaning (OR per day, 1.10; 95% CI, 1.06 1.13), chronic pulmonary disease other than chronic obstructive pulmonary disease (OR,13.23; 95% CI, 3.44-51.05), pneumonia as the reason to start mechanical ventilation (OR, 1.82; 95% CI, 1.07-3.08), and level of positive end-expiratory pressure applied before weaning (OR per unit,1.09; 95% CI, 1.04-1.14). The prolonged weaning group had a nonsignificant trend toward a higher rate of reintubation (P 1/4 0.08),tracheostomy (P 1/4 0.15), and significantly longer length of stay and higher mortality in the intensive care unit (OR for death, 1.97;95%CI, 1.17-3.31). The adjusted probability of death remained constant until Day 7, at which point it increased to 12.1%. PMID- 21617009 TI - Cellular therapy with sequential unmanipulated donor lymphocyte infusions in drug resistant cytomegalovirus (CMV) encephalitis. PMID- 21617010 TI - CD9 up-regulation on CD34+ cells with ingenol 3,20-dibenzoate does not improve homing in NSG mice. PMID- 21617011 TI - Fluorescence in situ hybridization confirms the presence of Merkel cell polyomavirus in chronic lymphocytic leukemia cells. PMID- 21617012 TI - The JAK2 46/1 haplotype in splanchnic vein thrombosis. PMID- 21617013 TI - West Nile virus infection transmitted by granulocyte transfusion. PMID- 21617014 TI - Permissive, nonpermissive HLA-DPB1 epitope disparities and the specificity of T cells infiltrating the skin during acute graft-versus-host disease. PMID- 21617016 TI - Population pharmacokinetic/pharmacodynamic modeling of clopidogrel in Korean healthy volunteers and stroke patients. AB - Population pharmacokinetic (PK) and pharmacodynamic (PD) modeling of clopidogrel was developed from pooled data from healthy volunteers (n = 44) and stroke patients (n = 35). The PK modeling used plasma concentrations of the clopidogrel metabolite (SR26334), and the PD modeling used platelet aggregation. The models were developed using NONMEM and evaluated via visual predictive check (VPC). Data were analyzed by 2-compartment modeling with Erlang's absorption and first-order elimination. There was no statistically significant covariate for each model parameter. The typical point estimates of PK were k(tr) (identical transfer rate constant) = 5.97 h(-1), k(e) (elimination rate constant) = 0.126 h(-1), k(d) (distribution rate constant) = 0.212 h(-1), V(2) (volume of central compartment) = 21.0 L, and V(3) (volume of peripheral compartment) = 38.8 L. The typical point estimates of PD were k(in) (input rate) = 27.9 h(-1), E(max) (maximum effect on input rate) = 0.292 h(-1), EC(5) (0) (median effective concentration) = 0.00629 ng/mL, and BASE (predose aggregation) = 66.7%. The final model was used to estimate individual parameters using patient data and showed good predictions using VPC. PMID- 21617017 TI - Is there an accurate method to measure metabolic requirement of institutionalized children with spastic cerebral palsy? AB - OBJECTIVES: This study hypothesized that there is no difference between energy expenditure measured by indirect calorimetry (IC) and that estimated by predicted formulas compared with the actual intake of children with spastic cerebral palsy (CP). METHODS: Fifteen children aged 3 to 18 years with spastic CP and associated complications were recruited. IC was used to measure mean energy expenditure (MEE) compared with 3 predicted equations for energy expenditure (PEE), including body surface area (BSA), the recommended daily allowance (RDA), and an equation designed specifically for patients with CP. Friedman and paired t tests were used to examine the variance between PEE and MEE. Intraclass correlation coefficient (ICC) was used to explore the correlation between MEE and PEE. The pretest and posttest core temperatures were compared using the Wilcoxon signed rank test. RESULTS: Mean +/- standard deviation MEE was 800.5 +/- 295.7 kcal/d; BSA was 1,213.4 +/- 171.2 kcal/d; RDA was 1,928.1 +/- 341.0 kcal/d; and CP was 1,603.1 +/ 215.8 kcal/d. The actual diet intake provided 935.3 +/- 222.9 kcal/d. Post hoc analysis revealed a significant difference between mean MEE and PEE (P < .001) but not mean actual intake (P = .128). In addition, the ICC of MEE vs PEE was 0.635 at a 95% confidence interval, indicating a weak correlation. In addition, mean pretest body temperature was 36.4 degrees C +/- 1 degrees C, and mean posttest body temperature was 35.8 degrees C +/- 2 degrees C. CONCLUSIONS: The study showed that MEE was significantly different from PEE, but not from actual intake. This warrants further exploration to develop a population-specific PEE for children with spastic CP. PMID- 21617018 TI - Activation of platelet membrane glycoprotein induced by parenteral nutrition: a preliminary report. AB - BACKGROUND: Long-term parenteral nutrition (PN) is a standardized treatment in many patients who are unable to use their gastrointestinal tract to absorb nutrients and water. Catheter-related thrombosis (CRT) is one of the most common complications of PN. Several factors predispose patients to CRT. The main objective of this study was to assess the platelet membrane glycoprotein (GP) activation and coagulopathy induced by PN. METHODS: Fifteen patients with intestinal failure were given PN for 47.9 days (range, 30-92 days) and 15 oral fed healthy volunteers served as controls. Complete blood counts and coagulation and biochemical parameters were determined. The platelet surface GPs, P-selectin and GPIIb/IIIa were measured by flow cytometry. RESULTS: There was no significant difference between the control group and PN group in coagulation and biochemical parameters. Platelet P-selectin expression of the PN group was significantly higher than that of the control group (3.43% +/- 1.22% and 1.99% +/- 0.58%, respectively; P < .01). There was no significant difference in GPIIb/IIIa expression between the 2 groups. CONCLUSIONS: Long-term PN (>30 days) induced the activation of platelet membrane GPs, which may be a significant risk factor for the development of CRT in patients with intestinal failure who require PN. PMID- 21617019 TI - Phase II trial of PTK787/ZK 222584 (vatalanib) administered orally once-daily or in two divided daily doses as second-line monotherapy in relapsed or progressing patients with stage IIIB/IV non-small-cell lung cancer (NSCLC). AB - BACKGROUND: The objective of this multicenter, prospective uncontrolled phase II trial was to determine efficacy, safety and tolerability of vatalanib, an oral angiogenesis inhibitor targeting all known vascular endothelial growth factor receptors, in the second-line treatment of non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: Patients with stage IIIB/IV NSCLC-proven tumor progression during or after one platinum-based chemotherapy regimen received a fixed dose of 1250 mg vatalanib either once-daily dosing (QD) or two divided daily dosing (TDD: 500 mg a.m. + 750 mg p.m.) until disease progression or unacceptable toxicity. Primary end point was the disease control rate (DCR) at 12 weeks. RESULTS: Fifty four and 58 patients were enrolled to the QD and TDD arms. DCR at 12 weeks was 35% in the QD and 37% in the TDD arm. The best overall response included one (2%) patient with confirmed partial response with QD and three (5%) with TDD. Median progression-free survival and overall survival were 2.1/7.3 months in the QD arm and 2.8/9.0 months with TDD arm. This therapy showed a moderate toxicity profile for the majority of patients. CONCLUSIONS: In the chosen patient population, vatalanib QD and TDD dosing demonstrated potential benefits in tumor size reduction, DCR, and survival. PMID- 21617020 TI - Dairy products and colorectal cancer risk: a systematic review and meta-analysis of cohort studies. AB - BACKGROUND: Previous studies of the association between intake of dairy products and colorectal cancer risk have indicated an inverse association with milk, however, the evidence for cheese or other dairy products is inconsistent. METHODS: We conducted a systematic review and meta-analysis to clarify the shape of the dose-response relationship between dairy products and colorectal cancer risk. We searched the PubMed database for prospective studies published up to May 2010. Summary relative risks (RRs) were estimated using a random effects model. RESULTS: Nineteen cohort studies were included. The summary RR was 0.83 (95% CI [confidence interval]: 0.78-0.88, I2=25%) per 400 g/day of total dairy products, 0.91 (95% CI: 0.85-0.94, I2=0%) per 200 g/day of milk intake and 0.96 (95% CI: 0.83-1.12, I2=28%) per 50 g/day of cheese. Inverse associations were observed in both men and women but were restricted to colon cancer. There was evidence of a nonlinear association between milk and total dairy products and colorectal cancer risk, P<0.001, and the inverse associations appeared to be the strongest at the higher range of intake. CONCLUSION: This meta-analysis shows that milk and total dairy products, but not cheese or other dairy products, are associated with a reduction in colorectal cancer risk. PMID- 21617021 TI - Effects of everyday romantic goal pursuit on women's attitudes toward math and science. AB - The present research examined the impact of everyday romantic goal strivings on women's attitudes toward science, technology, engineering, and math (STEM). It was hypothesized that women may distance themselves from STEM when the goal to be romantically desirable is activated because pursuing intelligence goals in masculine domains (i.e., STEM) conflicts with pursuing romantic goals associated with traditional romantic scripts and gender norms. Consistent with hypotheses, women, but not men, who viewed images (Study 1) or overheard conversations (Studies 2a-2b) related to romantic goals reported less positive attitudes toward STEM and less preference for majoring in math/science compared to other disciplines. On days when women pursued romantic goals, the more romantic activities they engaged in and the more desirable they felt, but the fewer math activities they engaged in. Furthermore, women's previous day romantic goal strivings predicted feeling more desirable but being less invested in math on the following day (Study 3). PMID- 21617022 TI - In vitro antifungal susceptibilities and molecular typing of sequentially isolated clinical Cryptococcus neoformans strains from Croatia. AB - A collection of 48 clinical Cryptococcus neoformans isolates from Croatia was investigated retrospectively using in vitro antifungal susceptibility testing and molecular biological techniques to determine mating type and serotype by PCR and amplified fragment length polymorphism (AFLP) genotyping. These isolates were obtained from 15 patients: ten were human immunodeficiency virus (HIV)-negative (66.7 %) and five were HIV-positive (33.3 %). From five patients, only one isolate was available, whilst from the other ten patients, two to 11 isolates were isolated sequentially. Antifungal susceptibility was tested by a broth microdilution method. Serotype A (genotype AFLP1) and serotype D (genotype AFLP2) were both found in six patients (40 % each), and serotype AD (genotype AFLP3) in three (20.0 %) patients. Mating type alpha (n = 12; 80.0 %) predominated and alpha/a hybrids were identified in 20.0 % of patients diagnosed with cryptococcosis. Two AFLP genotypes of C. neoformans were isolated during a single episode from one patient. The in vitro antifungal MIC(90) and susceptibility ranges for C. neoformans isolates were 0.5 ug ml(-1) (range 0.031-0.5 ug ml(-1)) for amphotericin B, 4 ug ml(-1) (range 1-4 ug ml(-1)) for flucytosine and fluconazole, 0.25 ug ml(-1) (range 0.031-0.5 ug ml(-1)) for itraconazole and 0.062 ug ml(-1) (range 0.031-0.25 ug ml(-1)) for voriconazole. PMID- 21617023 TI - The newly discovered ClpK protein strongly promotes survival of Klebsiella pneumoniae biofilm subjected to heat shock. PMID- 21617024 TI - Microarray-based detection of extended virulence and antimicrobial resistance gene profiles in phylogroup B2 Escherichia coli of human, meat and animal origin. AB - Extra-intestinal pathogenic Escherichia coli (ExPEC) causing urinary tract infections (UTIs) most often belong to phylogenetic group B2 and stem from the patient's own faecal flora. It has been hypothesized that the external reservoir for these uropathogenic E. coli in the human intestine may be meat and food production animals. To investigate such a connection, this study analysed an E. coli phylogroup B2 strain collection (n = 161) of geographical and temporally matched isolates, published previously, from UTI patients (n = 52), community dwelling humans (n = 36), imported (n = 5) and Danish (n = 13) broiler chicken meat, Danish broiler chickens (n = 17), imported (n = 3) and Danish (n = 27) pork, and healthy Danish pigs (n = 8). The isolates were subjected to microarray analysis for 315 virulence genes and variants and 82 antimicrobial resistance genes and variants. In total, 133 different virulence and antimicrobial resistance genes were detected in at least one UTI isolate. Between 66 and 87 of these genes were also detected in meat and animal isolates. Cluster analyses of virulence and resistance gene profiles, respectively, showed that UTI and community-dwelling human isolates most often grouped with meat and animal isolates, indicating genotypic similarity among such isolates. Furthermore, B2 isolates were detected from UTI patients and meat, with indistinguishable gene profiles. A considerable proportion of the animal and meat isolates belonged to the ExPEC pathotype. In conclusion, these findings suggest that B2 E. coli from meat and animal origin can be the source of most of the virulence and antimicrobial resistance genes detected in uropathogenic E. coli isolates and that there is a general resemblance of animal, meat and UTI E. coli based on extended gene profiling. These findings support the hypothesis of a zoonotic link between E. coli causing UTIs and E. coli from meat and animals. PMID- 21617028 TI - AtIPD: a curated database of Arabidopsis isoprenoid pathway models and genes for isoprenoid network analysis. PMID- 21617027 TI - Interfamily transfer of tomato Ve1 mediates Verticillium resistance in Arabidopsis. AB - Vascular wilts caused by soil-borne fungal species of the Verticillium genus are devastating plant diseases. The most common species, Verticillium dahliae and Verticillium albo-atrum, have broad host ranges and are notoriously difficult to control. Therefore, genetic resistance is the preferred method for disease control. Only from tomato (Solanum lycopersicum) has a Verticillium resistance locus been cloned, comprising the Ve1 gene that encodes a receptor-like protein type cell surface receptor. Due to lack of a suitable model for receptor-like protein (RLP)-mediated resistance signaling in Arabidopsis (Arabidopsis thaliana), so far relatively little is known about RLP signaling in pathogen resistance. Here, we show that Ve1 remains fully functional after interfamily transfer to Arabidopsis and that Ve1-transgenic Arabidopsis is resistant to race 1 but not to race 2 strains of V. dahliae and V. albo-atrum, nor to the Brassicaceae-specific pathogen Verticillium longisporum. Furthermore, we show that signaling components utilized by Ve1 in Arabidopsis to establish Verticillium resistance overlap with those required in tomato and include SERK3/BAK1, EDS1, and NDR1, which strongly suggests that critical components for resistance signaling are conserved. We subsequently investigated the requirement of SERK family members for Ve1 resistance in Arabidopsis, revealing that SERK1 is required in addition to SERK3/BAK1. Using virus-induced gene silencing, the requirement of SERK1 for Ve1-mediated resistance was confirmed in tomato. Moreover, we show the requirement of SERK1 for resistance against the foliar fungal pathogen Cladosporium fulvum mediated by the RLP Cf-4. Our results demonstrate that Arabidopsis can be used as model to unravel the genetics of Ve1 mediated resistance. PMID- 21617025 TI - A review of visual memory capacity: Beyond individual items and toward structured representations. AB - Traditional memory research has focused on identifying separate memory systems and exploring different stages of memory processing. This approach has been valuable for establishing a taxonomy of memory systems and characterizing their function but has been less informative about the nature of stored memory representations. Recent research on visual memory has shifted toward a representation-based emphasis, focusing on the contents of memory and attempting to determine the format and structure of remembered information. The main thesis of this review will be that one cannot fully understand memory systems or memory processes without also determining the nature of memory representations. Nowhere is this connection more obvious than in research that attempts to measure the capacity of visual memory. We will review research on the capacity of visual working memory and visual long-term memory, highlighting recent work that emphasizes the contents of memory. This focus impacts not only how we estimate the capacity of the system--going beyond quantifying how many items can be remembered and moving toward structured representations--but how we model memory systems and memory processes. PMID- 21617029 TI - Distinctive photosystem II photoinactivation and protein dynamics in marine diatoms. AB - Diatoms host chlorophyll a/c chloroplasts distinct from green chloroplasts. Diatoms now dominate the eukaryotic oceanic phytoplankton, in part through their exploitation of environments with variable light. We grew marine diatoms across a range of temperatures and then analyzed their PSII function and subunit turnover during an increase in light to mimic an upward mixing event. The small diatom Thalassiosira pseudonana initially responds to increased photoinactivation under blue or white light with rapid acceleration of the photosystem II (PSII) repair cycle. Increased red light provoked only modest PSII photoinactivation but triggered a rapid clearance of a subpool of PsbA. Furthermore, PsbD and PsbB content was greater than PsbA content, indicating a large pool of partly assembled PSII repair cycle intermediates lacking PsbA. The initial replacement rates for PsbD (D2) were, surprisingly, comparable to or higher than those for PsbA (D1), and even the supposedly stable PsbB (CP47) dropped rapidly upon the light shift, showing a novel aspect of rapid protein subunit turnover in the PSII repair cycle in small diatoms. Under sustained high light, T. pseudonana induces sustained nonphotochemical quenching, which correlates with stabilization of PSII function and the PsbA pool. The larger diatom Coscinodiscus radiatus showed generally similar responses but had a smaller allocation of PSII complexes relative to total protein content, with nearly equal stiochiometries of PsbA and PsbD subunits. Fast turnover of multiple PSII subunits, pools of PSII repair cycle intermediates, and photoprotective induction of nonphotochemical quenching are important interacting factors, particularly for small diatoms, to withstand and exploit high, fluctuating light. PMID- 21617030 TI - Phosphate deprivation in maize: genetics and genomics. PMID- 21617031 TI - Quantification of the brassinosteroid insensitive1 receptor in planta. AB - In plants, green fluorescent protein (GFP) is routinely used to determine the subcellular location of fusion proteins. Here, we show that confocal imaging can be employed to approximate the number of GFP-labeled protein molecules present in living Arabidopsis (Arabidopsis thaliana) root cells. The technique involves calibration with soluble GFP to provide a usable protein concentration range within the confocal volume of the microscope. As a proof of principle, we quantified the Brassinosteroid Insensitive1 (BRI1) receptor fused to GFP, under control of its own promoter. The number of BRI1-GFP molecules per root epidermal cell ranges from 22,000 in the meristem and 130,000 in the elongation zone to 80,000 in the maturation zone, indicating that up to 6-fold differences in BRI1 receptor content exist. In contrast, when taking into account differences in cell size, BRI1-GFP receptor density in the plasma membrane is kept constant at 12 receptors MUm-2 in all cells throughout the meristem and elongation zone. Only the quiescent center and columella cells deviate from this pattern and have 5 to 6 receptors MUm-2. Remarkably, root cell sensitivity toward brassinosteroids appears to coincide with uniform meristem receptor density. PMID- 21617032 TI - The thick aleurone1 mutant defines a negative regulation of maize aleurone cell fate that functions downstream of defective kernel1. AB - The maize (Zea mays) aleurone layer occupies the single outermost layer of the endosperm. The defective kernel1 (dek1) gene is a central regulator required for aleurone cell fate specification. dek1 mutants have pleiotropic phenotypes including lack of aleurone cells, aborted embryos, carotenoid deficiency, and a soft, floury endosperm deficient in zeins. Here we describe the thick aleurone1 (thk1) mutant that defines a novel negative function in the regulation of aleurone differentiation. Mutants possess multiple layers of aleurone cells as well as aborted embryos. Clonal sectors of thk1 mutant tissue in otherwise normal endosperm showed localized expression of the phenotype with sharp boundaries, indicating a localized cellular function for the gene. Sectors in leaves showed expanded epidermal cell morphology but the mutant epidermis generally remained in a single cell layer. Double mutant analysis indicated that the thk1 mutant is epistatic to dek1 for several aspects of the pleiotropic dek1 phenotype. dek1 mutant endosperm that was mosaic for thk1 mutant sectors showed localized patches of multilayered aleurone. Localized sectors were surrounded by halos of carotenoid pigments and double mutant kernels had restored zein profiles. In sum, loss of thk1 function restored the ability of dek1 mutant endosperm to accumulate carotenoids and zeins and to differentiate aleurone. Therefore the thk1 mutation defines a negative regulator that functions downstream of dek1 in the signaling system that controls aleurone specification and other aspects of endosperm development. The thk1 mutation was found to be caused by a deletion of approximately 2 megabases. PMID- 21617034 TI - Safety aspects of electroacupuncture. PMID- 21617035 TI - 'Forbidden points' in pregnancy: no plausible mechanism for risk. AB - It has been suggested that acupuncture may pose particular risks during pregnancy: by enhancing oxygenation to the developing embryo (presumably via increasing blood flow to the uterus); by affecting the level of maternal progesterone in early pregnancy; or by stimulating uterine contractions. This article examines the proposed risks and fails to find any plausible physiological mechanism for them. PMID- 21617036 TI - Skin changes after manual or electrical acupuncture. AB - Pigmentation of the skin after electroacupuncture (EA) or acupuncture in patients is not well recognised. Reporting and studying skin changes after acupuncture or EA would increase awareness among health services providers. This case series includes four cases of short-lived or long-term skin pigmentation after EA or acupuncture. Skin pigmentation or dark spots appeared in these four cases: after 12 treatments in Case 4, during the 14 treatments in Case 3, after one treatment course (16 treatments) in Case 1 and during the sixth month in a long-term treatment of duration 2 years (one or two treatments per week) in Case 2. These skin changes faded gradually after cessation of a course of acupuncture or EA. Skin pigmentation was hardly seen in one case after 14 days, and in another case after 42 days. Pigmentation of the skin could still be seen in one case after 42 days and could still be seen in another case after 2 years, however, in both cases, pigmentation has reduced in size and colour intensity by more than 50%. It is likely that pigmentation of the skin is a specific consequence of needling and may be closely linked to acupuncture dosage regardless of whether stimulation is given manually or by EA. PMID- 21617037 TI - Report of adverse event with electroacupuncture. AB - Electroacupuncture (EA) is becoming more common as a treatment for chronic musculoskeletal pain. It can be associated with adverse events related to the small electric currents used, in addition to the adverse events related to needle penetration of tissues. This paper reports a case of minor tissue damage following high intensity EA for 30 min with a device delivering a waveform that does not appear to be completely charge-balanced. This case highlights a rare but preventable adverse event. Manufacturers should be encouraged to develop EA stimulators that use charge-balanced waveforms. PMID- 21617039 TI - High pre-eruptive water contents preserved in lunar melt inclusions. AB - The Moon has long been thought to be highly depleted in volatiles such as water, and indeed published direct measurements of water in lunar volcanic glasses have never exceeded 50 parts per million (ppm). Here, we report in situ measurements of water in lunar melt inclusions; these samples of primitive lunar magma, by virtue of being trapped within olivine crystals before volcanic eruption, did not experience posteruptive degassing. The lunar melt inclusions contain 615 to 1410 ppm water and high correlated amounts of fluorine (50 to 78 ppm), sulfur (612 to 877 ppm), and chlorine (1.5 to 3.0 ppm). These volatile contents are very similar to primitive terrestrial mid-ocean ridge basalts and indicate that some parts of the lunar interior contain as much water as Earth's upper mantle. PMID- 21617040 TI - TFEB links autophagy to lysosomal biogenesis. AB - Autophagy is a cellular catabolic process that relies on the cooperation of autophagosomes and lysosomes. During starvation, the cell expands both compartments to enhance degradation processes. We found that starvation activates a transcriptional program that controls major steps of the autophagic pathway, including autophagosome formation, autophagosome-lysosome fusion, and substrate degradation. The transcription factor EB (TFEB), a master gene for lysosomal biogenesis, coordinated this program by driving expression of autophagy and lysosomal genes. Nuclear localization and activity of TFEB were regulated by serine phosphorylation mediated by the extracellular signal-regulated kinase 2, whose activity was tuned by the levels of extracellular nutrients. Thus, a mitogen-activated protein kinase-dependent mechanism regulates autophagy by controlling the biogenesis and partnership of two distinct cellular organelles. PMID- 21617042 TI - Selective attention from voluntary control of neurons in prefrontal cortex. AB - Animals can learn to voluntarily control neuronal activity within various brain areas through operant conditioning, but the relevance of that control to cognitive functions is unknown. We found that rhesus monkeys can control the activity of neurons within the frontal eye field (FEF), an oculomotor area of the prefrontal cortex. However, operantly driven FEF activity was primarily associated with selective visual attention, and not oculomotor preparation. Attentional effects were untrained and were observed both behaviorally and neurophysiologically. Furthermore, selective attention correlated with voluntary, but not spontaneous, fluctuations in FEF activity. Our results reveal a specific association of voluntarily driven neuronal activity with "top-down" attention and suggest a basis for the use of neurofeedback training to treat disorders of attention. PMID- 21617041 TI - Phosphorylation of the autophagy receptor optineurin restricts Salmonella growth. AB - Selective autophagy can be mediated via receptor molecules that link specific cargoes to the autophagosomal membranes decorated by ubiquitin-like microtubule associated protein light chain 3 (LC3) modifiers. Although several autophagy receptors have been identified, little is known about mechanisms controlling their functions in vivo. In this work, we found that phosphorylation of an autophagy receptor, optineurin, promoted selective autophagy of ubiquitin-coated cytosolic Salmonella enterica. The protein kinase TANK binding kinase 1 (TBK1) phosphorylated optineurin on serine-177, enhancing LC3 binding affinity and autophagic clearance of cytosolic Salmonella. Conversely, ubiquitin- or LC3 binding optineurin mutants and silencing of optineurin or TBK1 impaired Salmonella autophagy, resulting in increased intracellular bacterial proliferation. We propose that phosphorylation of autophagy receptors might be a general mechanism for regulation of cargo-selective autophagy. PMID- 21617043 TI - Innovation needs novel thinking. PMID- 21617048 TI - Cancer research. Texas's $3 billion fund lures scientific heavyweights. PMID- 21617049 TI - Immunology. Regulatory T cells get their chance to shine. PMID- 21617050 TI - Remote sensing. India's Earth-observing system comes under fire. PMID- 21617051 TI - Scientific community. The prion heretic. PMID- 21617052 TI - Bio-inspired engineering. Manta machines. PMID- 21617053 TI - The Biology of Genomes. On the trail of brain domestication genes. PMID- 21617054 TI - The Biology of Genomes. DNA circles cause cow coat color changes. PMID- 21617055 TI - The Biology of Genomes. Disease risk links to gene regulation. PMID- 21617056 TI - CT risks dwarfed by diagnostic benefits. PMID- 21617057 TI - Springtime for science in Egypt. PMID- 21617058 TI - Comment on "A persistent oxygen anomaly reveals the fate of spilled methane in the deep Gulf of Mexico". AB - Kessler et al. (Reports, 21 January 2011, p. 312) reported that methane released from the 2010 Deepwater Horizon blowout, approximately 40% of the total hydrocarbon discharge, was consumed quantitatively by methanotrophic bacteria in Gulf of Mexico deep waters over a 4-month period. We find the evidence explicitly linking observed oxygen anomalies to methane consumption ambiguous and extension of these observations to hydrate-derived methane climate forcing premature. PMID- 21617059 TI - Value of small forest fragments to amphibians. PMID- 21617060 TI - Oceans. Mitigating local causes of ocean acidification with existing laws. PMID- 21617062 TI - Physics. A logical use for atoms. PMID- 21617061 TI - Development. Limb cells don't tell time. PMID- 21617063 TI - Physics. Chameleon magnets. PMID- 21617064 TI - Behavior. Explaining human behavioral diversity. PMID- 21617065 TI - Biochemistry. KSR plays CRAF-ty. PMID- 21617066 TI - Geophysics. Assessing the status of Alaska's glaciers. PMID- 21617067 TI - SPORE series winner. The periodic table of videos. PMID- 21617068 TI - Dyscalculia: from brain to education. AB - Recent research in cognitive and developmental neuroscience is providing a new approach to the understanding of dyscalculia that emphasizes a core deficit in understanding sets and their numerosities, which is fundamental to all aspects of elementary school mathematics. The neural bases of numerosity processing have been investigated in structural and functional neuroimaging studies of adults and children, and neural markers of its impairment in dyscalculia have been identified. New interventions to strengthen numerosity processing, including adaptive software, promise effective evidence-based education for dyscalculic learners. PMID- 21617069 TI - Pure reasoning in 12-month-old infants as probabilistic inference. AB - Many organisms can predict future events from the statistics of past experience, but humans also excel at making predictions by pure reasoning: integrating multiple sources of information, guided by abstract knowledge, to form rational expectations about novel situations, never directly experienced. Here, we show that this reasoning is surprisingly rich, powerful, and coherent even in preverbal infants. When 12-month-old infants view complex displays of multiple moving objects, they form time-varying expectations about future events that are a systematic and rational function of several stimulus variables. Infants' looking times are consistent with a Bayesian ideal observer embodying abstract principles of object motion. The model explains infants' statistical expectations and classic qualitative findings about object cognition in younger babies, not originally viewed as probabilistic inferences. PMID- 21617070 TI - Experimental repetitive quantum error correction. AB - The computational potential of a quantum processor can only be unleashed if errors during a quantum computation can be controlled and corrected for. Quantum error correction works if imperfections of quantum gate operations and measurements are below a certain threshold and corrections can be applied repeatedly. We implement multiple quantum error correction cycles for phase-flip errors on qubits encoded with trapped ions. Errors are corrected by a quantum feedback algorithm using high-fidelity gate operations and a reset technique for the auxiliary qubits. Up to three consecutive correction cycles are realized, and the behavior of the algorithm for different noise environments is analyzed. PMID- 21617071 TI - Electrically induced ferromagnetism at room temperature in cobalt-doped titanium dioxide. AB - The electric field effect in ferromagnetic semiconductors enables switching of the magnetization, which is a key technology for spintronic applications. We demonstrated electric field-induced ferromagnetism at room temperature in a magnetic oxide semiconductor, (Ti,Co)O(2), by means of electric double-layer gating with high-density electron accumulation (>10(14) per square centimeter). By applying a gate voltage of a few volts, a low-carrier paramagnetic state was transformed into a high-carrier ferromagnetic state, thereby revealing the considerable role of electron carriers in high-temperature ferromagnetism and demonstrating a route to room-temperature semiconductor spintronics. PMID- 21617072 TI - Seismic imaging of transition zone discontinuities suggests hot mantle west of Hawaii. AB - The Hawaiian hotspot is often attributed to hot material rising from depth in the mantle, but efforts to detect a thermal plume seismically have been inconclusive. To investigate pertinent thermal anomalies, we imaged with inverse scattering of SS waves the depths to seismic discontinuities below the Central Pacific, which we explain with olivine and garnet transitions in a pyrolitic mantle. The presence of an 800- to 2000-kilometer-wide thermal anomaly (DeltaT(max) ~300 to 400 kelvin) deep in the transition zone west of Hawaii suggests that hot material does not rise from the lower mantle through a narrow vertical plume but accumulates near the base of the transition zone before being entrained in flow toward Hawaii and, perhaps, other islands. This implies that geochemical trends in Hawaiian lavas cannot constrain lower mantle domains directly. PMID- 21617073 TI - Computational design of virus-like protein assemblies on carbon nanotube surfaces. AB - There is a general need for the engineering of protein-like molecules that organize into geometrically specific superstructures on molecular surfaces, directing further functionalization to create richly textured, multilayered assemblies. Here we describe a computational approach whereby the surface properties and symmetry of a targeted surface define the sequence and superstructure of surface-organizing peptides. Computational design proceeds in a series of steps that encode both surface recognition and favorable intersubunit packing interactions. This procedure is exemplified in the design of peptides that assemble into a tubular structure surrounding single-walled carbon nanotubes (SWNTs). The geometrically defined, virus-like coating created by these peptides converts the smooth surfaces of SWNTs into highly textured assemblies with long scale order, capable of directing the assembly of gold nanoparticles into helical arrays along the SWNT axis. PMID- 21617074 TI - Impact of Antarctic Circumpolar Current development on late Paleogene ocean structure. AB - Global cooling and the development of continental-scale Antarctic glaciation occurred in the late middle Eocene to early Oligocene (~38 to 28 million years ago), accompanied by deep-ocean reorganization attributed to gradual Antarctic Circumpolar Current (ACC) development. Our benthic foraminiferal stable isotope comparisons show that a large delta(13)C offset developed between mid-depth (~600 meters) and deep (>1000 meters) western North Atlantic waters in the early Oligocene, indicating the development of intermediate-depth delta(13)C and O(2) minima closely linked in the modern ocean to northward incursion of Antarctic Intermediate Water. At the same time, the ocean's coldest waters became restricted to south of the ACC, probably forming a bottom-ocean layer, as in the modern ocean. We show that the modern four-layer ocean structure (surface, intermediate, deep, and bottom waters) developed during the early Oligocene as a consequence of the ACC. PMID- 21617075 TI - Initiation of proximal-distal patterning in the vertebrate limb by signals and growth. AB - Two broad classes of models have been proposed to explain the patterning of the proximal-distal axis of the vertebrate limb (from the shoulder to the digit tips). Differentiating between them, we demonstrate that early limb mesenchyme in the chick is initially maintained in a state capable of generating all limb segments through exposure to a combination of proximal and distal signals. As the limb bud grows, the proximal limb is established through continued exposure to flank-derived signal(s), whereas the developmental program determining the medial and distal segments is initiated in domains that grow beyond proximal influence. In addition, the system we have developed, combining in vitro and in vivo culture, opens the door to a new level of analysis of patterning mechanisms in the limb. PMID- 21617076 TI - Diffusible signals, not autonomous mechanisms, determine the main proximodistal limb subdivision. AB - Vertebrate limbs develop three main proximodistal (PD) segments (upper arm, forearm, and hand) in a proximal-to-distal sequence. Despite extensive research into limb development, whether PD specification occurs through nonautonomous or autonomous mechanisms is not resolved. Heterotopic transplantation of intact and recombinant chicken limb buds identifies signals in the embryo trunk that proximalize distal limb cells to generate a complete PD axis. In these transplants, retinoic acid induces proximalization, which is counteracted by fibroblast growth factors from the distal limb bud; these related actions suggest that the first limb-bud PD regionalization results from the balance between proximal and distal signals. The plasticity of limb progenitor cell identity in response to diffusible signals provides a unifying view of PD patterning during vertebrate limb development and regeneration. PMID- 21617077 TI - Differences between tight and loose cultures: a 33-nation study. AB - With data from 33 nations, we illustrate the differences between cultures that are tight (have many strong norms and a low tolerance of deviant behavior) versus loose (have weak social norms and a high tolerance of deviant behavior). Tightness-looseness is part of a complex, loosely integrated multilevel system that comprises distal ecological and historical threats (e.g., high population density, resource scarcity, a history of territorial conflict, and disease and environmental threats), broad versus narrow socialization in societal institutions (e.g., autocracy, media regulations), the strength of everyday recurring situations, and micro-level psychological affordances (e.g., prevention self-guides, high regulatory strength, need for structure). This research advances knowledge that can foster cross-cultural understanding in a world of increasing global interdependence and has implications for modeling cultural change. PMID- 21617078 TI - Ischemic preconditioning of the muscle improves maximal exercise performance but not maximal oxygen uptake in humans. AB - Brief episodes of nonlethal ischemia, commonly known as "ischemic preconditioning" (IP), are protective against cell injury induced by infarction. Moreover, muscle IP has been found capable of improving exercise performance. The aim of the study was the comparison of standard exercise performances carried out in normal conditions with those carried out following IP, achieved by brief muscle ischemia at rest (RIP) and after exercise (EIP). Seventeen physically active, healthy male subjects performed three incremental, randomly assigned maximal exercise tests on a cycle ergometer up to exhaustion. One was the reference (REF) test, whereas the others were performed after the RIP and EIP sessions. Total exercise time (TET), total work (TW), and maximal power output (W(max)), oxygen uptake (VO(2max)), and pulmonary ventilation (VE(max)) were assessed. Furthermore, impedance cardiography was used to measure maximal heart rate (HR(max)), stroke volume (SV(max)), and cardiac output (CO(max)). A subgroup of volunteers (n = 10) performed all-out tests to assess their anaerobic capacity. We found that both RIP and EIP protocols increased in a similar fashion TET, TW, W(max), VE(max), and HR(max) with respect to the REF test. In particular, W(max) increased by ~ 4% in both preconditioning procedures. However, preconditioning sessions failed to increase traditionally measured variables such as VO(2max), SV(max,) CO(max), and anaerobic capacity(.) It was concluded that muscle IP improves performance without any difference between RIP and EIP procedures. The mechanism of this effect could be related to changes in fatigue perception. PMID- 21617079 TI - Absence of meal-induced insulin sensitization (AMIS) in aging rats is associated with cardiac dysfunction that is protected by antioxidants. AB - We have previously demonstrated that progressive development of absence of meal induced insulin sensitization (AMIS) leads to postprandial hyperglycemia, compensatory hyperinsulinemia, resultant hyperlipidemia, increased oxidative stress, and obesity, progressing to syndrome X in aging rats. The present study tested the hypothesis that progressive development of AMIS in aging rats further resulted in deterioration in cardiac performance. Anesthetized male Sprague Dawley rats were tested at 9, 26, and 52 wk to determine their dynamic response to insulin and cardiac function. Dynamic insulin sensitivity was determined before and after atropine to quantitate hepatic insulin sensitizing substance (HISS)-dependent and -independent insulin action. Cardiac performance was evaluated using a Millar pressure-volume conductance catheter system. AMIS developed with age, as demonstrated by significant decrease in HISS-dependent insulin action, and this syndrome was increased by sucrose supplementation and inhibited by the antioxidant treatment. Associated with progressive development of AMIS, aging rats showed impaired cardiac performance, including the reduction in cardiac index, heart rate, dP/dt(max), dP/dt(min), ejection fraction and decreased slope of left ventricular end-systolic pressure-volume relationship, and increased relaxation time constant of left ventricular pressure as well as increased left ventricular end-diastolic pressure. Total peripheral vascular resistance also increased with age. Sucrose supplementation and antioxidant treatment, respectively, potentiated and attenuated cardiac dysfunction associated with age. In addition, poor cardiac performance correlated closely with the development of AMIS. These results indicate that AMIS is the first metabolic defect that leads to homeostatic disturbances and dysfunctions, including cardiovascular diseases. PMID- 21617080 TI - Echocardiographic detection of congestive heart failure in postinfarction rats. AB - In studies of congestive heart failure (CHF) treatment, it is essential to select animals with a similar degree of cardiac dysfunction. However, this is difficult to establish without hemodynamic evaluation in rat postinfarction-induced CHF. This study aimed to diagnose CHF in long-term follow-up postinfarction rats using only echocardiographic criteria through a J-tree cluster analysis and Fisher's linear discriminant function. Two sets of sham and infarcted rats were studied. The first was used to perform cluster analysis and the second to prospectively validate the results. Six months after inducing myocardial infarction (MI), rats were subjected to transthoracic echocardiography. Infarct size was measured by histological analysis. Six echocardiographic variables were used in the cluster analysis: left ventricular (LV) systolic dimension, LV diastolic dimension-to body weight ratio, left atrial diameter-to-body weight ratio, LV posterior wall shortening velocity, E wave, and isovolumetric relaxation time. Cluster analysis joined the rats into one sham and two MI groups. One MI cluster had more severe anatomical and echocardiographic changes and was called MI with heart failure (MI/HF+, n = 24, infarct size: 42.7 +/- 5.8%). The other had less severe changes and was called MI without heart failure (MI/HF-, n = 11, infarct size: 32.3 +/- 9.9%; P < 0.001 vs. MI/HF+). Three rats with small infarct size (21.6 +/- 2.2%) presenting mild cardiac alterations were misallocated in the sham group. Fisher's linear discriminant function was built using these groups and used to prospectively classify additional groups of sham-operated (n = 20) and infarcted rats (n = 57) using the same echocardiographic parameters. The discriminant function therefore detected CHF with 100% specificity and 80% sensitivity considering allocation in MI/HF+ and sham group, and 100% specificity and 58.8% sensitivity considering MI/HF+ and MI/HF- groups, taking into account pathological criteria of CHF diagnosis. Echocardiographic analysis can be used to accurately predict congestive heart failure in postinfarction rats. PMID- 21617081 TI - Editor's note regarding Lundberg et al. Letter to the Editor. PMID- 21617082 TI - Increases in cerebrovascular impedance in older adults. AB - This study explored a novel method for measuring cerebrovascular impedance to quantify the relationship between pulsatile changes in cerebral blood flow (CBF) and arterial pressure. Arterial pressure in the internal or common carotid artery (applanation tonometry), CBF velocity in the middle cerebral artery (transcranial Doppler), and end-tidal CO(2) (capnography) were measured in six young (28 +/- 4 yr) and nine elderly subjects (70 +/- 6 yr). Transfer function method was used to estimate cerebrovascular impedance. Under supine resting conditions, CBF velocity was reduced in the elderly despite the fact that they had higher arterial pressure than young subjects. As expected, cerebrovascular resistance index was increased in the elderly. In both young and elderly subjects, impedance modulus was reduced gradually in the frequency range of 0.78-8 Hz. Phase was negative in the range of 0.78-4.3 Hz and fluctuated at high frequencies. Compared with the young, impedance modulus increased by 38% in the elderly in the range of 0.78-2 Hz and by 39% in the range of 2-4 Hz (P < 0.05). Moreover, increases in impedance were correlated with reductions in CBF velocity. Collectively, these findings demonstrate the feasibility of assessing cerebrovascular impedance using the noninvasive method developed in this study. The estimated impedance modulus and phase are similar to those observed in the systemic circulation and other vascular beds. Moreover, increases in impedance in the elderly suggest that arterial stiffening, besides changes in cerebrovascular resistance, contributes to reduction in CBF with age. PMID- 21617083 TI - Mechanism of beneficial effects of physical activity on atherosclerosis and coronary heart disease. PMID- 21617084 TI - Estimated glomerular filtration rate is a poor predictor of concentration for a broad range of uremic toxins. AB - BACKGROUND AND OBJECTIVES: The degree of chronic kidney disease (CKD) is currently expressed in terms of GFR, which can be determined directly or estimated according to different formulas on the basis of serum creatinine and/or cystatin C measurements (estimated GFR [eGFR]). The purpose of this study was to investigate whether eGFR values are representative for uremic toxin concentrations in patients with different degrees of CKD. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Associations between eGFR based on serum cystatin C and different uremic solutes (mol wt range 113 to 240 D; determined by colorimetry, HPLC, or ELISA) were evaluated in 95 CKD patients not on dialysis (CKD stage 2 to 5). The same analysis was also applied for six other eGFR formulas. RESULTS: There was a substantial disparity in fits among solutes. In linear regression, explained variance of eGFR was extremely low for most solutes, with eGFR > 0.4 only for creatinine. The other eGFR formulations gave comparably disappointing results with regard to their association to uremic solutes. Relative similarity in R(2) values per solute for the different eGFR values and the strong disparity in values between solutes suggest that the differences in R(2) are mainly due to discrepancies in solute handling apart from GFR. CONCLUSIONS: eGFR is poorly associated with concentrations of all studied uremic toxins in patients with different degrees of CKD, correlates differently with each individual solute, and can thus not be considered representative for evaluating the accumulation of solutes in the course of CKD. PMID- 21617085 TI - Pre-emptive eculizumab and plasmapheresis for renal transplant in atypical hemolytic uremic syndrome. AB - The case of a 12-year-old with a hybrid CFH/CFHL1 gene and atypical hemolytic uremic syndrome (aHUS) that had previously developed native kidney and then renal allograft loss is reported. This case illustrates the relatively common occurrence of renal loss from the late presentation of aHUS. Also presented is a protocol for the pre-emptive use of eculizumab and plasmapheresis as part of a renal transplant plan for the treatment of aHUS in patients deemed at high risk for recurrent disease. This protocol was a result of a multidisciplinary approach including adult and pediatric nephrology, transplant surgery, transfusion medicine, and infectious disease specialists. This protocol and the justifications and components of it can function as a guideline for the treatment of a group of children that have waited in limbo for the first U.S. transplant to open the door to this type of definitive care for this devastating disease. PMID- 21617086 TI - Is coronary artery calcification associated with vertebral bone density in nondialyzed chronic kidney disease patients? AB - BACKGROUND AND OBJECTIVES: Low bone mineral density and coronary artery calcification (CAC) are highly prevalent among chronic kidney disease (CKD) patients, and both conditions are strongly associated with higher mortality. The study presented here aimed to investigate whether reduced vertebral bone density (VBD) was associated with the presence of CAC in the earlier stages of CKD. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Seventy-two nondialyzed CKD patients (age 52 +/- 11.7 years, 70% male, 42% diabetics, creatinine clearance 40.4 +/- 18.2 ml/min per 1.73 m(2)) were studied. VBD and CAC were quantified by computed tomography. RESULTS: CAC > 10 Agatston units (AU) was observed in 50% of the patients (median 120 AU [interquartile range 32 to 584 AU]), and a calcification score >= 400 AU was found in 19% (736 [527 to 1012] AU). VBD (190 +/- 52 Hounsfield units) correlated inversely with age (r = -0.41, P < 0.001) and calcium score (r = -0.31, P = 0.01), and no correlation was found with gender, creatinine clearance, proteinuria, lipid profile, mineral parameters, body mass index, and diabetes. Patients in the lowest tertile of VBD had expressively increased calcium score in comparison to the middle and highest tertile groups. In the multiple logistic regression analysis adjusting for confounding variables, low VBD was independently associated with the presence of CAC. CONCLUSIONS: Low VBD was associated with CAC in nondialyzed CKD patients. The authors suggest that low VBD might constitute another nontraditional risk factor for cardiovascular disease in CKD. PMID- 21617087 TI - Spinal epidural abscess in hemodialysis patients: a case series and review. AB - Spinal epidural abscess (SEA) is a rare but potentially devastating condition. We noticed an increase in the number of cases of SEA, with the majority in hemodialysis (HD) patients. This prompted a retrospective chart review of all cases of SEA admitted from 2000 to 2005 and a literature search of similar cases. We identified 19 SEA cases treated at Long Island College Hospital during this 6 year period, of which six were on HD: four were dialyzed via catheter, one via arteriovenous fistula, and in one the method of dialysis was not documented. Four patients had bacteremia with Staphylococcus aureus. Four patients presented with paresis or paralysis; only one improved. The mortality rate was 33% (2/6). We found 30 other cases of SEA in patients on HD from the literature. These 36 HD cases were compared with 85 SEA cases that were not on HD (13 from our study and 72 described in two large case series). The mortality rate was noted to be much higher in HD patients (23% [6/26] versus 7% [6/85]). Neurologic deficit at presentation was noted in 47% (17/36) of HD patients versus 69% (59/85) of non-HD patients, but neurologic improvement was higher in non-HD patients (71% [42/59] versus 29% [5/17]). This is the largest literature review of SEA in patients on HD. When compared with non-HD patients, HD patients had a higher mortality rate and were less likely to improve neurologically. PMID- 21617088 TI - Active vitamin D and acute respiratory infections in dialysis patients. AB - BACKGROUND AND OBJECTIVES: Vitamin D has gained attention for its pleiotropic effects in areas other than bone metabolism, and the effects of vitamin D in preventing respiratory infections have been reported as one of its immunomodulating properties. This study assessed the preventive effect of vitamin D receptor activator (VDRA) on respiratory infections in dialysis patients. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Maintained Japanese hemodialysis patients (n = 508) were observed for 5 years, and the incidence of hospitalization during this period because of acute respiratory infection (ARI) was recorded. RESULTS: Of the 508 patients, 212 had taken oral VDRA at the start of the study, whereas 296 patients had not received it. During the 5-year follow up period, 57 patients were hospitalized because of ARIs. Kaplan-Meier analysis revealed that the incidence of hospitalization because of respiratory infection was significantly lower in patients who had been treated with VDRA compared with patients who had not (log rank test; P = 0.02). The multivariate Cox proportional hazards model demonstrated that the patients who had taken oral VDRA were at a significantly lower risk of hospitalization because of respiratory disease (hazard ratio 0.47, 95% confidence interval 0.25 to 0.90). CONCLUSIONS: The findings of this study suggest that the administration of oral VDRA has a preventive effect on the incidence of ARIs in dialysis patients. PMID- 21617089 TI - Nurse-coordinated care in CKD: time for translation into practice? PMID- 21617090 TI - A nurse-coordinated model of care versus usual care for stage 3/4 chronic kidney disease in the community: a randomized controlled trial. AB - BACKGROUND AND OBJECTIVES: It is unclear how to optimally care for chronic kidney disease (CKD). This study compares a new coordinated model to usual care for CKD. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: A randomized trial in nephrology clinics and the community included 474 patients with median estimated GFR (eGFR) 42 ml/min per 1.73 m(2) identified by laboratory-based case finding compared care coordinated by a general practitioner (controls) with care by a nurse-coordinated team including a nephrologist (intervention) for a median (interquartile range [IQR]) of 742 days. 32% were diabetic, 60% had cardiovascular disease, and proteinuria was minimal. Guided by protocols, the intervention team targeted risk factors for adverse kidney and cardiovascular outcomes. Serial eGFR and clinical events were tracked. RESULTS: The average decline in eGFR over 20 months was -1.9 ml/min per 1.73 m(2). eGFR declined by >=4 ml/min per 1.73 m(2) within 20 months in 28 (17%) intervention patients versus 23 (13.9%) control patients. Control of BP, LDL, and diabetes were comparable across groups. In the intervention group there was a trend to greater use of renin-angiotensin blockers and more use of statins in those with initial LDL >2.5 mmol/L. Treatment was rarely required for anemia, acidosis, or disordered mineral metabolism. Clinical events occurred in 5.2% per year. CONCLUSIONS: Patients with stage 3/4 CKD identified through community laboratories largely had nonprogressive kidney disease but had cardiovascular risk. Over a median of 24 months, the nurse-coordinated team did not affect rate of GFR decline or control of most risk factors compared with usual care. PMID- 21617091 TI - Cost-effectiveness analysis of a randomized trial comparing care models for chronic kidney disease. AB - BACKGROUND AND OBJECTIVES: Potential cost and effectiveness of a nephrologist/nurse-based multifaceted intervention for stage 3 to 4 chronic kidney disease are not known. This study examines the cost-effectiveness of a chronic disease management model for chronic kidney disease. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Cost and cost-effectiveness were prospectively gathered alongside a multicenter trial. The Canadian Prevention of Renal and Cardiovascular Endpoints Trial (CanPREVENT) randomized 236 patients to receive usual care (controls) and another 238 patients to multifaceted nurse/nephrologist supported care that targeted factors associated with development of kidney and cardiovascular disease (intervention). Cost and outcomes over 2 years were examined to determine the incremental cost-effectiveness of the intervention. Base-case analysis included disease-related costs, and sensitivity analysis included all costs. RESULTS: Consideration of all costs produced statistically significant differences. A lower number of days in hospital explained most of the cost difference. For both base-case and sensitivity analyses with all costs included, the intervention group required fewer resources and had higher quality of life. The direction of the results was unchanged to inclusion of various types of costs, consideration of payer or societal perspective, changes to the discount rate, and levels of GFR. CONCLUSIONS: The nephrologist/nurse-based multifaceted intervention represents good value for money because it reduces costs without reducing quality of life for patients with chronic kidney disease. PMID- 21617092 TI - Converting structural information into an allosteric-energy-based picture for elongation factor Tu activation by the ribosome. AB - The crucial process of aminoacyl-tRNA delivery to the ribosome is energized by the GTPase reaction of the elongation factor Tu (EF-Tu). Advances in the elucidation of the structure of the EF-Tu/ribosome complex provide the rare opportunity of gaining a detailed understanding of the activation process of this system. Here, we use quantitative simulation approaches and reproduce the energetics of the GTPase reaction of EF-Tu with and without the ribosome and with several key mutants. Our study provides a novel insight into the activation process. It is found that the critical H84 residue is not likely to behave as a general base but rather contributes to an allosteric effect, which includes a major transition state stabilization by the electrostatic effect of the P loop and other regions of the protein. Our findings have general relevance to GTPase activation, including the processes that control signal transduction. PMID- 21617094 TI - System-level feedbacks make the anaphase switch irreversible. AB - The mitotic checkpoint prevents a eukaryotic cell from commencing to separate its replicated genome into two daughter cells (anaphase) until all of its chromosomes are properly aligned on the metaphase plate, with the two copies of each chromosome attached to opposite poles of the mitotic spindle. The mitotic checkpoint is exquisitely sensitive in that a single unaligned chromosome, 1 of a total of ~50, is sufficient to delay progression into anaphase; however, when the last chromosome comes into alignment on the metaphase plate, the mitotic checkpoint is quickly satisfied, and the replicated chromosomes are rapidly partitioned to opposite poles of the dividing cell. The mitotic checkpoint is also curious in the sense that, before metaphase alignment, chromosomes that are not being pulled in opposite directions by the mitotic spindle activate the checkpoint, but during anaphase, these same tensionless chromosomes can no longer activate the checkpoint. These and other puzzles associated with the mitotic checkpoint are addressed by a proposed molecular mechanism, which involves two positive feedback loops that create a bistable response of the checkpoint to chromosomal tension. PMID- 21617095 TI - The demise of "Nutcracker Man". PMID- 21617096 TI - Retinol-binding protein 4 is associated with prediabetes in adults from the general population: the Cooperative Health Research in the Region of Augsburg (KORA) F4 Study. AB - OBJECTIVE: We examined the association between retinol-binding protein 4 (RBP4), a novel adipokine, and prediabetes (isolated impaired fasting glucose [i-IFG], isolated impaired glucose tolerance [i-IGT], and combined IFG and IGT) in men and women aged 32-81 years. RESEARCH DESIGN AND METHODS: The analysis was based on 2,614 participants without previously diagnosed diabetes and those with newly diagnosed diabetes of the Cooperative Health Research in the Region of Augsburg (KORA) F4 Study, conducted from 2006 to 2008 in southern Germany. Plasma RBP4 was analyzed by immunonephelometry. RESULTS: In logistic regression analysis, RBP4 levels in the fourth quartile versus the first quartile were significantly associated with prediabetes (i-IGT, i-IFG, and IFG/IGT; reference normal glucose tolerance) independent of known metabolic risk factors and lifestyle variables (odds ratio 1.63 [95% CI 1.17-2.27] after multivariable adjustment). Stratification by sex showed generally similar results. CONCLUSIONS: RBP4 levels were associated with prediabetes in individuals from the general population. Prospective studies investigating the impact of RBP4 on the development of glucose intolerance are needed. PMID- 21617097 TI - Heterogeneity of altered cytokine levels across the clinical spectrum of diabetes in China. AB - OBJECTIVE: To determine the relationship between selected cytokines and diabetes in Chinese subjects. RESEARCH DESIGN AND METHODS: Adult patients with recent onset type 1 diabetes (n=53), latent autoimmune diabetes in adults (LADA) (n=250), and type 2 diabetes (n=285) from multiple centers were compared with normal subjects (n=196). We centrally tested serum GAD antibodies (GADAs), interleukin-6 (IL-6), lipocalin 2 (LCN2), high-sensitivity C-reactive protein (hs CRP), and adiponectin. RESULTS: After adjustment for age, sex, and BMI, all diabetes types had increased IL-6 and LCN2 (P<0.01), and all four cytokines were increased in LADA (P<0.01). In type 1 diabetes, adiponectin but not hs-CRP was increased (P<0.01), whereas in type 2 diabetes, hs-CRP but not adiponectin was increased (P<0.01). Adiponectin was correlated positively with GADA titer and negatively with hs-CRP (P<0.01 for both). CONCLUSIONS: In China, inflammatory markers are increased in all three major types of diabetes, but probably for different reasons, even in autoimmune diabetes. PMID- 21617098 TI - Salsalate attenuates free fatty acid-induced microvascular and metabolic insulin resistance in humans. AB - OBJECTIVE: Insulin recruits muscle microvasculature, thereby increasing endothelial exchange surface area. Free fatty acids (FFAs) cause insulin resistance by activating inhibitor of kappaB kinase beta. Elevating plasma FFAs impairs insulin's microvascular and metabolic actions in vivo. Whether salsalate, an anti-inflammatory agent, prevents FFA-induced microvascular and/or metabolic insulin resistance in humans is unknown. RESEARCH DESIGN AND METHODS: Eleven healthy, young adults were studied three times in random order. After an overnight fast, on two occasions each subject received a 5-h systemic infusion of Intralipid +/- salsalate pretreatment (50 mg/kg/day for 4 days). On the third occasion, saline replaced Intralipid. A 1 mU/kg/min euglycemic insulin clamp was superimposed over the last 2-h of each study. Skeletal and cardiac muscle microvascular blood volume (MBV), microvascular flow velocity (MFV), and microvascular blood flow (MBF) were determined before and after insulin infusion. Whole body glucose disposal rates were calculated from glucose infusion rates. RESULTS: Insulin significantly increased skeletal and cardiac muscle MBV and MBF without affecting MFV. Lipid infusion abolished insulin-mediated microvascular recruitment in both skeletal and cardiac muscle and lowered insulin-stimulated whole body glucose disposal (P<0.001). Salsalate treatment rescued insulin's actions to recruit muscle microvasculature and improved insulin-stimulated whole body glucose disposal in the presence of high plasma FFAs. CONCLUSIONS: High plasma concentrations of FFAs cause both microvascular and metabolic insulin resistance, which can be prevented or attenuated by salsalate treatment. Our data suggest that treatments aimed at inhibition of inflammatory response might help alleviate vascular insulin resistance and improve metabolic control in patients with diabetes. PMID- 21617099 TI - Insulin concerns and promises. PMID- 21617100 TI - Diabetic ketoacidosis complicated with previously unknown Gitelman syndrome in a Tunisian child. PMID- 21617101 TI - Diabetic ketosis caused by the insulin analog aspart-induced anti-insulin antibody: successful treatment with the newest insulin analog glulisine. PMID- 21617102 TI - Increasing the low-glucose alarm of a continuous glucose monitoring system prevents exercise-induced hypoglycemia without triggering any false alarms. PMID- 21617103 TI - Comment on: Lipsky and Berendt. Hyperbaric oxygen therapy for diabetic foot wounds: has hope hurdled hype? Diabetes Care 2010;33:1143-1145. PMID- 21617105 TI - Response to comment on Londahl et al: hyperbaric oxygen therapy facilitates healing of chronic foot ulcers in patients with diabetes. Diabetes Care 2010;33:998-1003. PMID- 21617106 TI - Comment on: Olsson et al. High levels of education are associated with an increased risk of latent autoimmune diabetes in adults: results from the Nord Trondelag Health Study. Diabetes Care 2011;34:102-107. PMID- 21617109 TI - Globalization of diabetes: the role of diet, lifestyle, and genes. AB - Type 2 diabetes is a global public health crisis that threatens the economies of all nations, particularly developing countries. Fueled by rapid urbanization, nutrition transition, and increasingly sedentary lifestyles, the epidemic has grown in parallel with the worldwide rise in obesity. Asia's large population and rapid economic development have made it an epicenter of the epidemic. Asian populations tend to develop diabetes at younger ages and lower BMI levels than Caucasians. Several factors contribute to accelerated diabetes epidemic in Asians, including the "normal-weight metabolically obese" phenotype; high prevalence of smoking and heavy alcohol use; high intake of refined carbohydrates (e.g., white rice); and dramatically decreased physical activity levels. Poor nutrition in utero and in early life combined with overnutrition in later life may also play a role in Asia's diabetes epidemic. Recent advances in genome-wide association studies have contributed substantially to our understanding of diabetes pathophysiology, but currently identified genetic loci are insufficient to explain ethnic differences in diabetes risk. Nonetheless, interactions between Westernized diet and lifestyle and genetic background may accelerate the growth of diabetes in the context of rapid nutrition transition. Epidemiologic studies and randomized clinical trials show that type 2 diabetes is largely preventable through diet and lifestyle modifications. Translating these findings into practice, however, requires fundamental changes in public policies, the food and built environments, and health systems. To curb the escalating diabetes epidemic, primary prevention through promotion of a healthy diet and lifestyle should be a global public policy priority. PMID- 21617110 TI - Influence of time of day of blood pressure-lowering treatment on cardiovascular risk in hypertensive patients with type 2 diabetes. AB - OBJECTIVE: We prospectively investigated in hypertensive patients with type 2 diabetes if bedtime treatment with >=1 hypertension medications exerts better blood pressure control and cardiovascular risk reduction than conventional therapy, in which all medications are ingested in the morning. RESEARCH DESIGN AND METHODS: We conducted a prospective, randomized, open-label, blinded end point trial on 448 hypertensive patients with type 2 diabetes, 255 men/193 women, mean +/- SD age 62.5 +/- 10.8 years, randomized to ingest all their prescribed hypertension medications upon awakening or >=1 of them at bedtime. Ambulatory blood pressure was measured for 48 h at baseline and again annually or even more frequently (quarterly) after adjustments in treatment. RESULTS: After a median follow-up of 5.4 years, patients ingesting >=1 hypertension medications at bedtime showed a significantly lower cardiovascular risk (adjusted by age and sex) than subjects ingesting all medications upon awakening (hazard ratio 0.33 [95% CI 0.21-0.54]; P < 0.001). The difference between groups in the adjusted risk of major events (cardiovascular death, myocardial infarction, and stroke) was also statistically significant (0.25 [0.10-0.61]; P = 0.003). Patients treated at bedtime showed significantly lower sleep time blood pressure mean and higher prevalence of controlled ambulatory blood pressure (62.5 vs. 50.9%; P = 0.013). There was a significant 12% cardiovascular risk reduction per each 5 mmHg decrease in asleep systolic blood pressure during follow-up (P < 0.001). CONCLUSIONS: Among patients with diabetes, treatment with >=1 hypertension medications at bedtime, compared with all medications upon waking, resulted in improved ambulatory blood pressure control and significantly reduced cardiovascular morbidity and mortality. PMID- 21617111 TI - Position statement executive summary: guidelines and recommendations for laboratory analysis in the diagnosis and management of diabetes mellitus. AB - BACKGROUND: Multiple laboratory tests are used in the diagnosis and management of patients with diabetes mellitus. The quality of the scientific evidence supporting the use of these assays varies substantially. APPROACH: An expert committee compiled evidence-based recommendations for the use of laboratory analysis in patients with diabetes. A new system was developed to grade the overall quality of the evidence and the strength of the recommendations. A draft of the guidelines was posted on the Internet, and the document was modified in response to comments. The guidelines were reviewed by the joint Evidence-Based Laboratory Medicine Committee of the AACC and the National Academy of Clinical Biochemistry and were accepted after revisions by the Professional Practice Committee and subsequent approval by the Executive Committee of the American Diabetes Association. CONTENT: In addition to the long-standing criteria based on measurement of venous plasma glucose, diabetes can be diagnosed by demonstrating increased hemoglobin A(1c) (HbA(1c)) concentrations in the blood. Monitoring of glycemic control is performed by the patients measuring their own plasma or blood glucose with meters and by laboratory analysis of HbA(1c). The potential roles of noninvasive glucose monitoring, genetic testing, and measurement of autoantibodies, urine albumin, insulin, proinsulin, C-peptide, and other analytes are addressed. SUMMARY: The guidelines provide specific recommendations based on published data or derived from expert consensus. Several analytes are found to have minimal clinical value at the present time, and measurement of them is not recommended. PMID- 21617112 TI - Use of metformin in the setting of mild-to-moderate renal insufficiency. PMID- 21617108 TI - Guidelines and recommendations for laboratory analysis in the diagnosis and management of diabetes mellitus. AB - BACKGROUND: Multiple laboratory tests are used to diagnose and manage patients with diabetes mellitus. The quality of the scientific evidence supporting the use of these tests varies substantially. APPROACH: An expert committee compiled evidence-based recommendations for the use of laboratory testing for patients with diabetes. A new system was developed to grade the overall quality of the evidence and the strength of the recommendations. Draft guidelines were posted on the Internet and presented at the 2007 Arnold O. Beckman Conference. The document was modified in response to oral and written comments, and a revised draft was posted in 2010 and again modified in response to written comments. The National Academy of Clinical Biochemistry and the Evidence-Based Laboratory Medicine Committee of the American Association for Clinical Chemistry jointly reviewed the guidelines, which were accepted after revisions by the Professional Practice Committee and subsequently approved by the Executive Committee of the American Diabetes Association. CONTENT: In addition to long-standing criteria based on measurement of plasma glucose, diabetes can be diagnosed by demonstrating increased blood hemoglobin A(1c) (HbA(1c)) concentrations. Monitoring of glycemic control is performed by self-monitoring of plasma or blood glucose with meters and by laboratory analysis of HbA(1c). The potential roles of noninvasive glucose monitoring, genetic testing, and measurement of autoantibodies, urine albumin, insulin, proinsulin, C-peptide, and other analytes are addressed. SUMMARY: The guidelines provide specific recommendations that are based on published data or derived from expert consensus. Several analytes have minimal clinical value at present, and their measurement is not recommended. PMID- 21617113 TI - Chronobiology impacts response to antihypertensive drug regimen in type 2 diabetes. PMID- 21617115 TI - Specific epidermal growth factor receptor autophosphorylation sites promote mouse colon epithelial cell chemotaxis and restitution. AB - Upon ligand binding, epidermal growth factor (EGF) receptor (R) autophosphorylates on COOH-terminal tyrosines, generating docking sites for signaling partners that stimulate proliferation, restitution, and chemotaxis. Specificity for individual EGFR tyrosines in cellular responses has been hypothesized but not well documented. Here we tested the requirement for particular tyrosines, and associated downstream pathways, in mouse colon epithelial cell chemotactic migration. We compared these requirements to those for the phenotypically distinct restitution (wound healing) migration. Wild-type, Y992/1173F, Y1045F, Y1068F, and Y1086F EGFR constructs were expressed in EGFR(-/ ) cells; EGF-induced chemotaxis or restitution were determined by Boyden chamber or modified scratch wound assay, respectively. Pharmacological inhibitors of p38, phospholipase C (PLC), Src, MEK, JNK/SAPK, phosphatidylinositol 3-kinase (PI 3 kinase), and protein kinase C (PKC) were used to block EGF-stimulated signaling. Pathway activation was determined by immunoblot analysis. Unlike wild-type EGFR, Y992/1173F and Y1086F EGFR did not stimulate colon epithelial cell chemotaxis toward EGF; Y1045F and Y1068F EGFR partially stimulated chemotaxis. Only wild type EGFR promoted colonocyte restitution. Inhibition of p38, PLC, and Src, or Grb2 knockdown, blocked chemotaxis; JNK, PI 3-kinase, and PKC inhibitors or c-Cbl knockdown blocked restitution but not chemotaxis. All four EGFR mutants stimulated downstream signaling in response to EGF, but Y992/1173F EGFR was partially defective in PLCgamma activation whereas both Y1068F and Y1086F EGFR failed to activate Src. We conclude that specific EGFR tyrosines play key roles in determining cellular responses to ligand. Chemotaxis and restitution, which have different migration phenotypes and physiological consequences, have overlapping but not identical EGFR signaling requirements. PMID- 21617116 TI - Acidic bile salts modulate the squamous epithelial barrier function by modulating tight junction proteins. AB - Experimental models for esophageal epithelium in vitro either suffer from poor differentiation or complicated culture systems. An air-liquid interface system with normal human bronchial epithelial cells can serve as a model of esophageal like squamous epithelial cell layers. Here, we explore the influence of bile acids on barrier function and tight junction (TJ) proteins. The cells were treated with taurocholic acid (TCA), glycocholic acid (GCA), or deoxycholic acid (DCA) at different pH values, or with pepsin. Barrier function was measured by transepithelial electrical resistance (TEER) and the diffusion of paracellular tracers (permeability). The expression of TJ proteins, including claudin-1 and claudin-4, was examined by Western blotting of 1% Nonidet P-40-soluble and insoluble fractions. TCA and GCA dose-dependently decreased TEER and increased paracellular permeability at pH 3 after 1 h. TCA (4 mM) or GCA (4 mM) did not change TEER and permeability at pH 7.4 or pH 4. The combination of TCA and GCA at pH 3 significantly decreased TEER and increased permeability at lower concentrations (2 mM). Pepsin (4 mg/ml, pH 3) did not have any effect on barrier function. DCA significantly decreased the TEER and increased permeability at pH 6, a weakly acidic condition. TCA (4 mM) and GCA (4 mM) significantly decreased the insoluble fractions of claudin-1 and claudin-4 at pH 3. In conclusion, acidic bile salts disrupted the squamous epithelial barrier function partly by modulating the amounts of claudin-1 and claudin-4. These results provide new insights for understanding the role of TJ proteins in esophagitis. PMID- 21617118 TI - Trends in the timing of pre-emptive kidney transplantation. AB - Pre-emptive kidney transplantation is considered the best available renal replacement therapy, but no guidelines exist to direct its timing during CKD progression. We used a national cohort of 19,471 first-time pre-emptive kidney transplant recipients between 1995-2009 to evaluate patterns and implications of transplant timing. Mean estimated GFR (eGFR) at the time of pre-emptive transplant increased significantly over time, from 9.2 ml/min/1.73 m(2) in 1995 to 13.8 ml/min/1.73 m(2) in 2009 (P<0.001). Patients with eGFR >= 15 ml/min/1.73 m(2) represented an increasing proportion of pre-emptive transplant recipients, from 9% in 1995 to 35% in 2009; the trend for patients with eGFR >= 10 was similar (30% to 72%). We did not detect statistically significant differences in patient survival or death-censored graft survival between strata of eGFR at the time of transplant, either in the full cohort or in subgroup analyses of patients who might theoretically benefit from earlier pre-emptive transplantation. In summary, pre-emptive kidney transplantation is occurring at increasing levels of native kidney function. Earlier transplantation does not appear to associate with patient or graft survival, suggesting that earlier pre-emptive transplantation may subject donors and recipients to premature operative risk and waste the native kidney function of recipients. PMID- 21617119 TI - Minding the gap: connexin 40 at the heart of renin release. PMID- 21617117 TI - TNF-alpha converting enzyme-mediated ErbB4 transactivation by TNF promotes colonic epithelial cell survival. AB - Disruption of intestinal epithelial homeostasis, including enhanced apoptosis, is a hallmark of inflammatory bowel disease (IBD). We have recently shown that tumor necrosis factor (TNF) increases the kinase activity of ErbB4, a member of the epidermal growth factor receptor family that is elevated in mucosa of IBD patients and that promotes colon epithelial cell survival. In this study, we tested the hypothesis that TNF transactivates ErbB4 through TNF-alpha converting enzyme (TACE)-mediated ligand release and that this transactivation is necessary to protect colonic epithelial cells from cytokine-induced apoptosis. Using neutralizing antibodies, we show that heparin-binding EGF-like growth factor (HB EGF) is required for ErbB4 phosphorylation in response to TNF. Pharmacological or genetic inhibition of the metalloprotease TACE, which mediates HB-EGF release from cells, blocked TNF-induced ErbB4 activation. MEK, but not Src or p38, was also required for transactivation. TACE activity and ligand binding were required for ErbB4-mediated antiapoptotic signaling; whereas mouse colon epithelial cells expressing ErbB4 were resistant to TNF-induced apoptosis, TACE inhibition or blockade of ErbB4 ligand binding reversed the survival advantage. We conclude that TNF transactivates ErbB4 through TACE-dependent HB-EGF release, thus protecting colon epithelial cells from cytokine-induced apoptosis. These findings have important implications for understanding how ErbB4 protects the colon from apoptosis-induced tissue injury in inflammatory conditions such as IBD. PMID- 21617120 TI - Making sense of the latest advice on vitamin D therapy. AB - The Institute of Medicine recently published recommendations for the daily intake and optimal serum levels of vitamin D based on an extensive review of the existing literature. Here we examine the issue and put levels of vitamin D in context for the general population and in patients with chronic kidney disease. Large randomized controlled trials are necessary to ensure that current recommendations are appropriate. PMID- 21617121 TI - Suramin inhibits renal fibrosis in chronic kidney disease. AB - The activation of cytokine and growth factor receptors associates with the development and progression of renal fibrosis. Suramin is a compound that inhibits the interaction of several cytokines and growth factors with their receptors, but whether suramin inhibits the progression of renal fibrosis is unknown. Here, treatment of cultured renal interstitial fibroblasts with suramin inhibited their activation induced by TGF-beta1 and serum. In a mouse model of obstructive nephropathy, administration of a single dose of suramin immediately after ureteral obstruction abolished the expression of fibronectin, largely suppressed expression of alpha-SMA and type I collagen, and reduced the deposition of extracellular matrix proteins. Suramin also decreased the expression of multiple cytokines including TGF-beta1 and reduced the interstitial infiltration of leukocytes. Moreover, suramin decreased expression of the type II TGF-beta receptor, blocked phosphorylation of the EGF and PDGF receptors, and inactivated several signaling pathways associated with the progression of renal fibrosis. In a rat model of CKD, suramin abrogated proteinuria, limited the decline of renal function, and prevented glomerular and tubulointerstitial damage. Collectively, these findings indicate that suramin is a potent antifibrotic agent that may have therapeutic potential for patients with fibrotic kidney diseases. PMID- 21617122 TI - DRB1*15 allele is a risk factor for PR3-ANCA disease in African Americans. AB - Anti-neutrophil cytoplasmic autoantibody (ANCA) disease rarely occurs in African Americans and risk factors for the disease in this population are unknown. Here, we genotyped MHC class II alleles and found that, among African Americans, those with proteinase 3-ANCA (PR3-ANCA) had 73.3-fold higher odds of having HLA-DRB1*15 alleles than community-based controls (OR 73.3; 95% CI 9.1 to 591). In addition, a disproportionate number of African American patients carried the DRB1*1501 allelic variant of Caucasian descent rather than the DRB1*1503 allelic variant of African descent. Among Caucasians, those with PR3-ANCA had 2.2-fold higher odds of carrying DRB1*1501 than controls (OR 2.2; 95% CI 1.2 to 4.0). A validation study supported by the Vasculitis Clinical Research Consortium confirmed the strong association between the DRB1*15 allele and PR3-ANCA disease, among African Americans. Furthermore, we found that DRB1*1501 protein binds with high affinity to amino acid sequences of sense-PR3, purportedly an antigenic epitope, and to the amino acid sequence complementary to this epitope in vitro. Peptides of sense PR3 and complementary-PR3 also bound to TNF-alpha-induced surface expression of DRB1*1501 on peripheral neutrophils. Taken together, these data suggest HLA DRB1*15 alleles contribute to the pathogenesis of PR3-ANCA disease. PMID- 21617123 TI - Nilotinib attenuates renal injury and prolongs survival in chronic kidney disease. AB - The tyrosine kinase inhibitor imatinib is beneficial in experimental renal diseases, but the effect of the new tyrosine kinase inhibitor nilotinib on the progression of renal failure is unknown. We administered either nilotinib or vehicle to Sprague-Dawley rats beginning 2 weeks after 5/6 nephrectomy (Nx) or laparotomy and continuing for 8 weeks. Serum creatinine levels were significantly lower in the nilotinib group after 6 and 8 weeks of treatment. Furthermore, nilotinib-treated rats had less proteinuria, attenuated glomerulosclerosis and tubulointerstitial damage, and reduced macrophage infiltration into the tubulointerstitium. Treatment with nilotinib also significantly decreased renal cortical expression of profibrogenic genes, such as IL-1beta and monocyte chemotactic protein-1, which correlated closely with the tubulointerstitial damage score and ED1-positive macrophages score. In addition, nilotinib treatment significantly prolonged survival. Taken together, these results suggest that nilotinib may limit the progression of chronic kidney disease. PMID- 21617124 TI - Trials and tribulations of new agents, novel biomarkers, and retarding renal progression. PMID- 21617125 TI - REIN on obesity, proteinuria and CKD. PMID- 21617126 TI - Sulfatide-reactive natural killer T cells abrogate ischemia-reperfusion injury. AB - There is a significant immune response to ischemia-reperfusion injury (IRI), but the role of immunomodulatory natural killer T (NKT) cell subtypes is not well understood. Here, we compared the severity of IRI in mice deficient in type I/II NKT cells (CD1d(-/-)) or type I NKT cells (Jalpha18(-/-)). The absence of NKT cells, especially type II NKT cells, accentuated the severity of renal injury, whereas repletion of NKT cells attenuated injury. Adoptively transferred NKT cells trafficked into the tubulointerstitium, which is the primary area of injury. Sulfatide-induced activation of type II NKT cells protected kidneys from IRI, but inhibition of NKT cell recruitment enhanced injury. In co-culture experiments, sulfatide-induced activation of NKT cells from either mice or humans attenuated apoptosis of renal tubular cells after transient hypoxia via hypoxia inducible factor (HIF)-1alpha and IL-10 pathways. Renal tissue of patients with acute tubular necrosis (ATN) frequently contained NKT cells, and the number of these cells tended to negatively correlate with ATN severity. In summary, sulfatide-reactive type II NKT cells are renoprotective in IRI, suggesting that pharmacologic modulation of NKT cells may protect against ischemic injury. PMID- 21617127 TI - Correlates of physician visits among older adults in China: the effects of family support. AB - OBJECTIVE: We examined how family support influenced the use of health services among older Chinese adults. METHOD: Data came from a national representative survey including 20,255 respondents aged 60 and older. The dependent variable was the number of physician visits in the past 12 months. Family support variables include living arrangement, family size, financial support, instrumental support, and filial piety. RESULTS: Providing or receiving financial support increased the likelihood as well as number of physician visits. By contrast, living with children and regarding children as filial decreased physician visits. DISCUSSION: Financial sufficiency as indicated by the exchange of financial resources within families indicates the importance of money in predicting older adults' physician visits. Living with children may indicate a higher level of support, which substitutes some of physician services. Perceiving children as being filial may render psychological protective effects to older adults which results in less health service use. PMID- 21617128 TI - Interactions between ankyrin-G, Plakophilin-2, and Connexin43 at the cardiac intercalated disc. AB - RATIONALE: The early description of the intercalated disc defined 3 structures, all of them involved in cell-cell communication: desmosomes, gap junctions, and adherens junctions. Current evidence demonstrates that molecules not involved in providing a physical continuum between cells also populate the intercalated disc. Key among them is the voltage-gated sodium channel complex. An important component of this complex is the cytoskeletal adaptor protein Ankyrin-G (AnkG). OBJECTIVE: To test the hypothesis that AnkG partners with desmosome and gap junction molecules and exerts a functional effect on intercellular communication in the heart. METHODS AND RESULTS: We used a combination of microscopy, immunochemistry, patch-clamp, and optical mapping to assess the interactions between AnkG, Plakophilin-2, and Connexin43. Coimmunoprecipitation studies from rat heart lysate demonstrated associations between the 3 molecules. With the use of siRNA technology, we demonstrated that loss of AnkG expression caused significant changes in subcellular distribution and/or abundance of PKP2 and Connexin43 as well as a decrease in intercellular adhesion strength and electric coupling. Regulation of AnkG and of Na(v)1.5 by Plakophilin-2 was also demonstrated. Finally, optical mapping experiments in AnkG-silenced cells demonstrated a shift in the minimal frequency at which rate-dependence activation block was observed. CONCLUSIONS: These experiments support the hypothesis that AnkG is a key functional component of the intercalated disc at the intersection of 3 complexes often considered independent: the voltage-gated sodium channel, gap junctions, and the cardiac desmosome. Possible implications to the pathophysiology of inherited arrhythmias (such as arrhythmogenic right ventricular cardiomyopathy) are discussed. PMID- 21617129 TI - Atg7 induces basal autophagy and rescues autophagic deficiency in CryABR120G cardiomyocytes. AB - RATIONALE: Increasing evidence suggests that misfolded proteins and intracellular aggregates contribute to cardiac disease and heart failure. Several cardiomyopathies, including the alphaB-crystallin R120G mutation (CryAB(R120G)) model of desmin-related cardiomyopathy, accumulate cytotoxic misfolded proteins in the form of preamyloid oligomers and aggresomes. Impaired autophagic function is a potential cause of misfolded protein accumulations, cytoplasmic aggregate loads, and cardiac disease. Atg7, a mediator of autophagosomal biogenesis, is a putative regulator of autophagic function. OBJECTIVE: To determine whether autophagic induction by Atg7 is sufficient to reduce misfolded protein and aggregate content in protein misfolding-stressed cardiomyocytes. METHODS AND RESULTS: To define the gain and loss of function effects of Atg7 expression on CryAB(R120G) protein misfolding and aggregates, neonatal rat cardiomyocytes were infected with adenoviruses expressing either wild-type CryAB or CryAB(R120G) and coinfected with Atg7 adenovirus or with Atg7 silencing siRNAs to produce gain-of or loss-of Atg7 function. Atg7 overexpression effectively induced basal autophagy with no detrimental effects on cell survival, suggesting that Atg7 can activate autophagy with no apparent cytotoxic effects. Autophagic flux assays on CryAB(R120G)-expressing cardiomyocytes revealed reduced autophagic function, which probably contributed to the failure of misfolded proteins and aggregates to be cleared. Coexpression of Atg7 and CryAB(R120G) significantly reduced preamyloid oligomer staining, aggregate content, and cardiomyocyte cytotoxicity. Conversely, Atg7 silencing in the CryAB(R120G) background significantly inhibited the already reduced rate of autophagy and increased CryAB(R120G) aggregate content and cytotoxicity. CONCLUSIONS: Atg7 induces basal autophagy, rescues the CryAB(R120G) autophagic deficiency, and attenuates the accumulation of misfolded proteins and aggregates in cardiomyocytes. PMID- 21617130 TI - Endogenous muscle atrophy F-box mediates pressure overload-induced cardiac hypertrophy through regulation of nuclear factor-kappaB. AB - RATIONALE: Overexpression of muscle atrophy F-box (MAFbx/atrogin-1), an E3 ubiquitin ligase, induces proteasomal degradation in cardiomyocytes. The role of endogenous MAFbx in regulating cardiac hypertrophy and failure remains unclear. OBJECTIVE: We investigated the role of MAFbx in regulating cardiac hypertrophy and function in response to pressure overload. Transverse aortic constriction (TAC) was applied to MAFbx knockout (KO) and wild-type (WT) mice. METHODS AND RESULTS: Expression of MAFbx in WT mice was significantly increased by TAC. TAC induced increases in cardiac hypertrophy were significantly smaller in MAFbx KO than in WT mice. There was significantly less lung congestion and interstitial fibrosis in MAFbx KO than in WT mice. MAFbx KO also inhibited beta-adrenergic cardiac hypertrophy. DNA microarray analysis revealed that activation of genes associated with the transcription factor binding site for the nuclear factor kappaB family were inhibited in MAFbx KO mice compared with WT mice after TAC. Although the levels of IkappaB-alpha were significantly decreased after TAC in WT mice, they were increased in MAFbx KO mice. MAFbx regulates ubiquitination and proteasomal degradation of IkappaB-alpha in cardiomyocytes. In primary cultured rat cardiomyocytes, phenylephrine-induced activation of nuclear factor-kappaB and hypertrophy were significantly suppressed by MAFbx knockdown but were partially rescued by overexpression of nuclear factor-kappaB p65. CONCLUSIONS: MAFbx plays an essential role in mediating cardiac hypertrophy in response to pressure overload. Downregulation of MAFbx inhibits cardiac hypertrophy in part through stabilization of IkappaB-alpha and inactivation of nuclear factor-kappaB. Taken together, inhibition of MAFbx attenuates pathological hypertrophy, thereby protecting the heart from progression into heart failure. PMID- 21617131 TI - Novel role of the CXC chemokine receptor 3 in inflammatory response to arterial injury: involvement of mTORC1: retraction. PMID- 21617133 TI - The ontogeny of cardiac regeneration. PMID- 21617132 TI - What is the oncologic risk of stem cell treatment for heart disease? PMID- 21617134 TI - Thematic series on the pathobiology of vascular calcification: an introduction. AB - Vascular calcification increasingly afflicts our aging, dysmetabolic population. Once considered only a passive process of dead and dying cells, data from multiple laboratories worldwide have converged to demonstrate that vascular calcification is a highly regulated form of biomineralization. The goal of this thematic review series is to highlight what is known concerning the biological "players" and "game rules" with respect to vascular mineral metabolism. Armed with this understanding, it is hoped that novel therapeutic strategies can be crafted to prevent and treat vascular calcium accrual, to the benefit of our patients afflicted with arteriosclerotic valvular and vascular diseases. PMID- 21617135 TI - Molecular imaging insights into early inflammatory stages of arterial and aortic valve calcification. AB - Traditional imaging modalities such as computed tomography, although perfectly adept at identifying and quantifying advanced calcification, cannot detect the early stages of this disorder and offer limited insight into the mechanisms of mineral dysregulation. This review presents optical molecular imaging as a promising tool that simultaneously detects pathobiological processes associated with inflammation and early stages of calcification in vivo at the (sub)cellular levels. Research into treatment of cardiovascular calcification is lacking, as shown by clinical trials that have failed to demonstrate the reduction of calcific aortic stenosis. Hence, the need to elucidate the pathways that contribute to cardiovascular calcification and to develop new therapeutic strategies to prevent or reverse calcification has driven investigations into the use of molecular imaging. This review discusses studies that have used molecular imaging methods to advance knowledge of cardiovascular calcification, focusing in particular on the inflammation-dependent mechanisms of arterial and aortic valve calcification. PMID- 21617137 TI - Aging causes collateral rarefaction and increased severity of ischemic injury in multiple tissues. AB - OBJECTIVE: Aging is a major risk factor for increased ischemic tissue injury. Whether collateral rarefaction and impaired remodeling contribute to this is unknown. We quantified the number and diameter of native collaterals and their remodeling in 3-, 16-, 24-, and 31-month-old mice. METHODS AND RESULTS: Aging caused an "age-dose-dependent" greater drop in perfusion immediately after femoral artery ligation, followed by a diminished recovery of flow and increase in tissue injury. These effects were associated with a decline in collateral number, diameter, and remodeling. Angiogenesis was also impaired. Mechanistically, these changes were not accompanied by reduced recruitment of T cells or macrophages to remodeling collaterals. However, endothelial nitric oxide synthase signaling was dysfunctional, as indicated by increased protein nitrosylation and less phosphorylated endothelial nitric oxide synthase and vasodilator-stimulated phosphoprotein in collateral wall cells. The cerebral circulation exhibited a similar age-dose-dependent loss of collateral number and diameter and increased tortuosity, resulting in an increase in collateral resistance and infarct volume (eg, 6- and 3-fold, respectively, in 24-month-old mice) after artery occlusion. This was not associated with rarefaction of similarly sized arterioles. Collateral remodeling was also reduced. CONCLUSIONS: Our findings demonstrate that aging causes rarefaction and insufficiency of the collateral circulation in multiple tissues, resulting in more severe ischemic tissue injury. PMID- 21617138 TI - Reduced thrombin formation and altered fibrin clot properties induced by polyunsaturated omega-3 fatty acids on top of dual antiplatelet therapy in patients undergoing percutaneous coronary intervention (OMEGA-PCI clot). AB - OBJECTIVE: The goal of this study was to investigate whether omega-3 polyunsaturated fatty acids (n-3 PUFA) are able to alter plasma fibrin clot properties and reduce thrombin formation in stable coronary artery disease patients undergoing percutaneous coronary intervention (PCI). METHODS AND RESULTS: In an investigator-initiated, prospective, double-blind, placebo controlled, randomized study, patients undergoing PCI who received standard pharmacotherapy were assigned to the treatment with 1 g/day n-3 PUFA (n = 30) or placebo (n = 24) for 1 month. Plasma fibrin clot permeability (K(s)); lysis time (t(50%)); prothrombin fragment 1.2; and peak thrombin generation from automated thrombogram, 8-isoprostaglandin F(2alpha) (8-iso-PGF(2alpha), an oxidative stress marker), and C-reactive protein were determined at baseline, 3 to 5 days after randomization, and 30 days after randomization. At baseline, both treatment groups did not differ significantly. A 1-month treatment with n-3 PUFA compared with placebo was associated with 15.3% higher K(s), indicating larger pores in the fibrin network (P = 0.0005); 14.3% shorter t(50%), indicating increased susceptibility to fibrinolysis (P<0.0001); 33.8% lower prothrombin fragment 1.2 (P = 0.0013); 13.4% lower peak thrombin generation (P = 0.04); and 13.1% lower 8 iso-PGF(2alpha) (P = 0.009). Treatment with n-3 PUFA had no effect on fibrinogen and C-reactive protein. After 1 month of treatment, fibrinogen (r = -0.53, P<0.0001), treatment assignment (r = 0.29, P = 0.006) and 8-iso-PGF(2alpha) (r = 0.27, P = 0.015) were independently associated with clot permeability (P<0.0001, R(2) = 0.66). CONCLUSIONS: Adding n-3 PUFA to standard therapy in stable patients undergoing PCI significantly decreases thrombin formation and oxidative stress and favorably alters fibrin clot properties. These findings indicate novel antithrombotic effects induced by n-3 PUFA in humans. PMID- 21617139 TI - Inhibition of pathological differentiation of valvular interstitial cells by C type natriuretic peptide. AB - OBJECTIVE: Calcific aortic valve disease is associated with the differentiation of valvular interstitial cells (VICs) to myofibroblast and osteoblast-like cells, particularly in the fibrosa layer of the valve. Previous studies suggested that C type natriuretic peptide (CNP) protects against calcific aortic valve disease to maintain homeostasis. We aimed to determine whether CNP inhibits VIC pathological differentiation as a mechanism to explain its protective effects. METHODS AND RESULTS: CNP expression was prominent in normal porcine aortic valves, particularly on the ventricular side, but reduced in sclerotic valves concomitant with the appearance of pathological VIC phenotypes in the fibrosa. In vitro, CNP inhibited calcified aggregate formation and bone-related transcript and protein expression by VICs grown in osteogenic conditions. Under myofibrogenic culture conditions, CNP reduced alpha-smooth muscle actin expression and cell-mediated gel contraction, indicating inhibition of myofibroblast differentiation. Similar to CNP, simvastatin inhibited VIC osteoblast and myofibroblast differentiation in vitro. Strikingly, simvastatin upregulated CNP expression in VICs cultured under myofibrogenic conditions, and small interfering RNA knockdown of natriuretic peptide receptor-b (a CNP receptor) significantly reduced the antifibrotic effect of simvastatin, suggesting that it acts in part via CNP/NPR-B autocrine/paracrine signaling. CONCLUSIONS: CNP inhibits myofibroblast and osteoblast differentiation of VICs and is responsible in part for inhibition of VIC myofibroblast differentiation by statins, suggesting novel mechanisms to explain the protective effect of CNP and the pleiotropic effects of statins in the aortic valve. PMID- 21617136 TI - Calcific aortic valve stenosis: methods, models, and mechanisms. AB - Calcific aortic valve stenosis (CAVS) is a major health problem facing aging societies. The identification of osteoblast-like and osteoclast-like cells in human tissue has led to a major paradigm shift in the field. CAVS was thought to be a passive, degenerative process, whereas now the progression of calcification in CAVS is considered to be actively regulated. Mechanistic studies examining the contributions of true ectopic osteogenesis, nonosseous calcification, and ectopic osteoblast-like cells (that appear to function differently from skeletal osteoblasts) to valvular dysfunction have been facilitated by the development of mouse models of CAVS. Recent studies also suggest that valvular fibrosis, as well as calcification, may play an important role in restricting cusp movement, and CAVS may be more appropriately viewed as a fibrocalcific disease. High-resolution echocardiography and magnetic resonance imaging have emerged as useful tools for testing the efficacy of pharmacological and genetic interventions in vivo. Key studies in humans and animals are reviewed that have shaped current paradigms in the field of CAVS, and suggest promising future areas for research. PMID- 21617140 TI - Arginase I attenuates inflammatory cytokine secretion induced by lipopolysaccharide in vascular smooth muscle cells. AB - OBJECTIVE: Inflammation plays an important role in atherosclerosis. Arginase I (Arg I) promotes the proliferation of vascular smooth muscle cells; however, the effect of Arg I on inflammation remains unknown. The present study investigated the role of Arg I in inflammation in vitro and in vivo. METHODS AND RESULTS: Quantitative reverse transcription-polymerase chain reaction and Western blot analysis demonstrated that Arg I inhibited tumor necrosis factor-alpha production induced by lipopolysaccharide in human aortic smooth muscle cells. Inducible nitric oxide synthase substrate competition and nuclear factor-kappaB activation were main contributors to lipopolysaccharide-mediated inflammatory cytokine generation. However, Arg I could attenuate the function of inducible nitric oxide synthase and inhibit the subsequent nuclear factor-kappaB activation, leading to inhibition of tumor necrosis factor-alpha generation. Furthermore, upregulation of Arg I significantly decreased macrophage infiltration and inflammation in atherosclerotic plaque of rabbits, whereas downregulation of Arg I aggravated these adverse effects. CONCLUSIONS: The results indicate the antiinflammatory effects of Arg I and suggest an unexpected beneficial role of Arg I in inflammatory disease. PMID- 21617141 TI - Knockout of toll-like receptor-2 attenuates both the proinflammatory state of diabetes and incipient diabetic nephropathy. AB - OBJECTIVE: Type 1 diabetes (T1DM) is a proinflammatory state and confers an increased risk for vascular complications. Toll-like receptors (TLR) could participate in diabetic vasculopathies. Whether TLR activation contributes to the proinflammatory state of T1DM and the pathogenesis of diabetic nephropathy remains unknown. METHODS AND RESULTS: We induced T1DM in TLR2 knockout mice (TLR2 /-) and wild-type littermates (C57BL/6J-WT) using streptozotocin (STZ). Fasting blood, peritoneal macrophages, and kidneys were obtained for flow cytometry, Western blot, microscopy, and cytokine assays at 6 and 14 weeks after induction of diabetes. Macrophage TLR2 expression and MyD88-dependent signaling were increased in diabetic mice (WT+STZ) compared with nondiabetic WT mice. These biomarkers were attenuated in diabetic TLR2-/- macrophages. WT+STZ mice showed increased kidney:body weight ratio due to cell hypertrophy, increased albuminuria, decreased kidney nephrin, podocin, and podocyte number and increased transforming growth factor-beta and laminin compared with WT mice. Nephrin, podocin, and podocyte number and effacement were restored, and transforming growth factor-beta and laminin levels were decreased in TLR2-/-+ STZ mice kidneys versus WT+STZ. Peritoneal and kidney macrophages were predominantly M1 phenotype in WT+STZ mice; this was attenuated in TLR2-/-+STZ mice. CONCLUSIONS: These data support a role for TLR2 in promoting inflammation and early changes of incipient diabetic nephropathy, in addition to albuminuria and podocyte loss. PMID- 21617142 TI - Aldosterone regulates vascular gene transcription via oxidative stress-dependent and -independent pathways. AB - OBJECTIVE: Aldosterone (Aldo) antagonism prevents cardiovascular mortality by unclear mechanisms. Aldo binds to the mineralocorticoid receptor (MR), a ligand activated transcription factor, which is expressed in human vascular cells. Here we define the early Aldo-regulated vascular transcriptome and investigate the mechanisms of gene regulation by Aldo in the vasculature that may contribute to vascular disease. METHODS AND RESULTS: Gene expression profiling of Aldo-treated mouse aortas identified 72 genes regulated by Aldo. These genes are overrepresented in Gene Ontology categories involved in vascular function and disease. Quantitative reverse transcription-polymerase chain reaction was used to confirm and further explore mechanisms of vascular gene regulation by Aldo. Aldo regulated vascular gene expression was inhibited by actinomycin D and MR antagonists supporting a transcriptional MR-dependent mechanism. Aldo regulation of a subset of genes was enhanced in the setting of vascular endothelial denudation and blocked by the free radical scavenger Tempol, supporting synergy between Aldo and vascular injury that is oxidative stress dependent. In the aortic arch, a region predisposed to atherosclerosis, the injury-enhanced genes also demonstrated enhanced expression compared with the descending aorta, both at baseline and after Aldo exposure. Furthermore, the clinically beneficial MR antagonist spironolactone inhibited expression of the identified genes in aortic tissue from humans with atherosclerosis. CONCLUSIONS: This study defines the Aldo regulated vascular transcriptome and characterizes a subset of proatherogenic genes with enhanced Aldo-stimulated, oxidative stress-dependent expression in the setting of vascular injury and in areas predisposed to atherosclerosis. Inhibition of MR regulation of these genes may play a role in the protective effects of Aldo antagonists in patients with vascular disease, and these pathways may provide novel drug targets to prevent atherosclerosis in humans. PMID- 21617143 TI - Two aces: transient ischemic attack work-up as outpatient assessment of clinical evaluation and safety. AB - BACKGROUND AND PURPOSE: To evaluate a novel emergency department-based TIA triage system. METHODS: We developed an approach to TIA triage and management based on risk assessment using the ABCD(2) score in combination with early cervical and intracranial vessel imaging. It was anticipated that this triage system would avoid hospitalization for the majority of TIA patients and result in a low rate of recurrent stroke. We hypothesized that the subsequent stroke rate among consecutively encountered patients managed with this approach would be lower than predicted based on their ABCD2 scores. RESULTS: From June 2007 to December 2009, 224 consecutive patients evaluated in the Stanford emergency department for a possible TIA were enrolled in the study. One hundred fifty-seven were discharged to complete their evaluation at the outpatient TIA clinic; 67 patients were hospitalized. One hundred sixteen patients had a final diagnosis of TIA/minor stroke or possible TIA. The stroke rates at 7, 30, and 90 days were 0.6% (0.1% 3.5%) for patients referred to the TIA clinic and 1.5% (0.3%-8.0%) for the hospitalized patients. Combining both groups, the overall stroke rate was 0.9% (0.3%-3.2%), which is significantly less than expected based on ABCD2 scores (P=0.034 at 7 days and P=0.001 at 90 days). CONCLUSIONS: This emergency department-based inpatient versus outpatient TIA triage system led to a low rate of hospitalization (30%). Recurrent stroke rates were low for both the hospitalized and outpatient subgroups. PMID- 21617144 TI - Hypertension and cerebral vascular reactivity in childhood: challenge and opportunity. PMID- 21617145 TI - Impairment of cerebral blood flow regulation in astronauts with orthostatic intolerance after flight. AB - BACKGROUND AND PURPOSE: We investigated cerebral blood flow regulation in astronauts before and after flights. We hypothesized that autoregulation would be different before flight and after flight between nonfinishers and the finishers of a stand test. METHODS: Twenty-seven astronauts from shuttle missions lasting 8 to 16 days underwent a 10-minute stand test: 10 days before flight, 1 to 2 hours and 3 days after landing. Mean blood flow velocity of the middle cerebral artery (MCA) was measured using transcranial Doppler; Mean arterial pressure was measured using a Finapres (Ohmeda, Englewood, CO) and was adjusted to the level of the MCA (BP(MCA)). Cross-spectral power, gain, phase, and coherence were determined for the relation between BP(MCA) and the cerebrovascular resistance index mean blood flow velocity/BP(MCA). RESULTS: BP(MCA) was reduced with stand (P<0.001). Differences between finishers and nonfinishers (P=0.011) and over test days (P=0.004) were observed. Cerebrovascular conductance was affected by stand (P<0.001), by group (P<0.001) with a group by stand, and test day interaction (P<0.01). Preflight data suggest that the nonfinishers were operating at a higher cerebral vasodilation than finishers for a given BP(MCA), and on landing day the nonfinishers had a greater decrease in mean blood flow velocity as a function of BP(MCA) with standing compared to finishers and preflight. There was a significant interaction effect of gender over the test days and from supine to stand (P=0.035). CONCLUSIONS: Our results indicate that the cause of presyncope in astronauts may be related to a mismatch of cerebral blood flow with blood pressure. Astronaut gender may also play a role in susceptibility to orthostatic intolerance after flight. PMID- 21617146 TI - Cerebrospinal fluid 20-HETE is associated with delayed cerebral ischemia and poor outcomes after aneurysmal subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: Delayed cerebral ischemia (DCI) is a major complication after aneurysmal subarachnoid hemorrhage (aSAH); it is manifested by changes in cerebral blood flow accompanied by neurological decline, and it results in long term functional and neuropsychological impairment. Preclinical evidence has demonstrated that the arachidonic acid metabolite, 20-hydroxyeicosatetraenoic acid (20-HETE), affects cerebral microvascular tone and cerebral blood flow after aSAH. The purpose of this study was to determine whether cerebrospinal fluid 20 HETE levels were associated with DCI and long-term neuropsychological outcomes in aSAH patients. METHODS: Cerebrospinal fluid samples were collected twice daily through 14 days after hemorrhage on 108 acute, adult, aSAH patients. Samples were analyzed for 20-HETE via HPLC MSQ single quadrupole mass spectrometry. DCI was defined as the presence of impaired cerebral blood flow (angiographic vasospasm, elevated transcranial Dopplers, abnormal computed tomography or magnetic resonance perfusion scans) accompanied by neurological deterioration. Outcomes, including death and neuropsychological testing, were completed at 3 months after hemorrhage. RESULTS: Detectable 20-HETE levels were observed in 31% of patient samples and were associated with severity of hemorrhage (Hunt & Hess [HH], P=0.04; Fisher, P=0.05). Detection of 20-HETE was not associated with angiographic vasospasm (P=0.34); however, detectable 20-HETE was significantly associated with DCI (P=0.016). Our data also suggest that detectable 20-HETE was associated with decreased performance in 5 neuropsychological domains. CONCLUSIONS: These results provide the first clinical evidence that cerebrospinal fluid 20-HETE concentrations are associated with DCI and poor outcomes, and this provides impetus for future studies to elucidate the clinical utility of inhibiting 20-HETE formation as a novel therapeutic intervention in patients with aSAH. PMID- 21617147 TI - Racial-ethnic disparities in stroke care: the American experience: a statement for healthcare professionals from the American Heart Association/American Stroke Association. AB - PURPOSE: Our goal is to describe the effect of race and ethnicity on stroke epidemiology, personal beliefs, access to care, response to treatment, and participation in clinical research. In addition, we seek to determine the state of knowledge on the main factors that may explain disparities in stroke care, with the goal of identifying gaps in knowledge to guide future research. The intended audience includes physicians, nurses, other healthcare professionals, and policy makers. METHODS: Members of the writing group were appointed by the American Heart Association Stroke Council Scientific Statement Oversight Committee and represent different areas of expertise in relation to racial-ethnic disparities in stroke care. The writing group reviewed the relevant literature, with an emphasis on reports published since 1972. The statement was approved by the writing group; the statement underwent peer review, then was approved by the American Heart Association Science Advisory and Coordinating Committee. RESULTS: There are limitations in the definitions of racial and ethnic categories currently in use. For the purpose of this statement, we used the racial categories defined by the US federal government: white, black or African American, Asian, American Indian/Alaskan Native, and Native Hawaiian/other Pacific Islander. There are 2 ethnic categories: people of Hispanic/Latino origin or not of Hispanic/Latino origin. There are differences in the distribution of the burden of risk factors, stroke incidence and prevalence, and stroke mortality among different racial and ethnic groups. In addition, there are disparities in stroke care between minority groups compared with whites. These disparities include lack of awareness of stroke symptoms and signs and lack of knowledge about the need for urgent treatment and the causal role of risk factors. There are also differences in attitudes, beliefs, and compliance among minorities compared with whites. Differences in socioeconomic status and insurance coverage, mistrust of the healthcare system, the relatively limited number of providers who are members of minority groups, and system limitations may contribute to disparities in access to or quality of care, which in turn might result in different rates of stroke morbidity and mortality. Cultural and language barriers probably also contribute to some of these disparities. Minorities use emergency medical services systems less, are often delayed in arriving at the emergency department, have longer waiting times in the emergency department, and are less likely to receive thrombolysis for acute ischemic stroke. Although unmeasured factors may play a role in these delays, the presence of bias in the delivery of care cannot be excluded. Minorities have equal access to rehabilitation services, although they experience longer stays and have poorer functional status than whites. Minorities are inadequately treated with both primary and secondary stroke prevention strategies compared with whites. Sparse data exist on racial ethnic disparities in access to surgical care after intracerebral hemorrhage and subarachnoid hemorrhage. Participation of minorities in clinical research is limited. Barriers to participation in clinical research include beliefs, lack of trust, and limited awareness. Race is a contentious topic in biomedical research because race is not proven to be a surrogate for genetic constitution. CONCLUSIONS: There are limitations in the current definitions of race and ethnicity. Nevertheless, racial and ethnic disparities in stroke exist and include differences in the biological determinants of disease and disparities throughout the continuum of care, including access to and quality of care. Access to and participation in research is also limited among minority groups. Acknowledging the presence of disparities and understanding the factors that contribute to them are necessary first steps. More research is required to understand these differences and find solutions. PMID- 21617148 TI - Community-based participatory research: a new approach to engaging community members to rapidly call 911 for stroke. AB - BACKGROUND AND PURPOSE: Acute stroke treatments are underutilized primarily because of delayed hospital arrival. Using a community-based participatory research approach, we explored stroke self-efficacy, knowledge, and perceptions of stroke among a predominately African American population in Flint, Michigan. METHODS: In March 2010, a survey was administered to youth and adults after religious services at 3 churches and during 1 church health day. The survey consisted of vignettes (12 stroke, 4 nonstroke) to assess knowledge of stroke warning signs and behavioral intent to call 911. The survey also assessed stroke self-efficacy, personal knowledge of someone who had experienced a stroke, personal history of stroke, and barriers to calling 911. Linear regression models explored the association of stroke self-efficacy with behavioral intent to call 911 among adults. RESULTS: Two hundred forty-two adults and 90 youths completed the survey. Ninety-two percent of adults and 90% of youth respondents were African American. Responding to 12 stroke vignettes, adults would call 911 in 72% (SD, 0.26) of the vignettes, whereas youths would call 911 in 54% of vignettes (SD, 0.29; P<0.001). Adults correctly identified stroke in 51% (SD, 0.32) of the stroke vignettes and youth correctly identified stroke in 46% (SD, 0.28) of the stroke vignettes (P=0.28). Stroke self-efficacy predicted behavioral intent to call 911 (P=0.046). CONCLUSIONS: In addition to knowledge of stroke warning signs, behavioral interventions to increase both stroke self-efficacy and behavioral intent may be useful for helping people make appropriate 911 calls for stroke. A community-based participatory research approach may be effective in reducing stroke disparities. PMID- 21617149 TI - Hypertension impairs vascular reactivity in the pediatric brain. AB - BACKGROUND AND PURPOSE: Chronic hypertension impairs cerebrovascular regulation in adults, but its effects on the pediatric population are unknown. The objective of this study was to investigate cerebrovascular abnormalities in hypertensive children and adolescents. METHODS: Sixty-four children and adolescents aged 7 to 20 years underwent transcranial Doppler examinations of the middle cerebral artery at the time of rebreathing CO2. Time-averaged maximum mean cerebral blood flow velocity and end-tidal CO2 were used to quantify cerebrovascular reactivity during hypercapnia. Patients were clinically categorized as hypertensive, prehypertensive, or white coat hypertensive based on 24-hour ambulatory blood pressure measurements. Their reactivities were compared with 9 normotensive control subjects and evaluated against baseline mean blood pressure z-scores and loads. RESULTS: Untreated hypertensive children had significantly lower hypercapnic reactivity than normotensive children (2.556 +/- 1.832 cm/s x mm Hg versus 4.256 +/- 1.334 cm/s x mm Hg, P < 0.05). Baseline mean diastolic blood pressure z-scores (r = -0.331, P = 0.037) and diastolic blood pressure loads (r = -0.351, P = 0.026) were inversely related to reactivity. CONCLUSIONS: Untreated hypertensive children and adolescents have blunted reactivity to hypercapnia, indicating deranged vasodilatory reactivity. The inverse relationship between diastolic blood pressure indices and reactivity suggests that diastolic blood pressure may be a better predictor of cerebral end organ damage than systolic blood pressure. PMID- 21617150 TI - Outcomes after carotid artery stenting and endarterectomy in the Medicare population. AB - BACKGROUND AND PURPOSE: Carotid artery stenting (CAS) is an alternative to carotid endarterectomy (CEA) for stroke prevention. The value of this therapy relative to CEA remains uncertain. METHODS: In 10 958 Medicare patients aged 66 years or older between 2004 and 2006, we analyzed in-hospital, 1-year stroke, myocardial infarction, and death rate outcomes and the effects of potential confounding variables. RESULTS: CAS patients (87% were asymptomatic) had a higher baseline risk profile, including having a higher percentage of coronary and peripheral arterial disease, heart failure, and renal failure. In-hospital stroke rate (1.9% CAS versus 1.4% CEA; P=0.14) and mortality (CAS 0.9% versus 0.6% CEA; P=0.20) were similar. By 1 year, CAS patients had similar stroke rates (5.3% CAS versus 4.1% CEA; P=0.12) but higher all-cause mortality rates (9.9% CAS versus 6.1% CEA; P<0.001). Using Cox multivariable models, there was a similar stroke risk (hazard ratio, 1.28; 95% CI, 0.90-1.79) but CAS patients had a significantly higher mortality (HR, 1.32; 95% CI, 1.02-1.71). Sensitivity analyses suggested that unmeasured confounders could be responsible for the mortality difference. In multivariable analysis, stroke risk was highest in the patients symptomatic at the time of revascularization. CONCLUSIONS: CAS patients had a similar stroke risk but an increased mortality rate at 1 year compared with CEA patients, possibly related to the higher baseline risk profile in the CAS patient group. PMID- 21617151 TI - Letter by Lahr et al regarding article, "Promoting thrombolysis in acute ischemic stroke". PMID- 21617152 TI - Guidelines and recommendations for laboratory analysis in the diagnosis and management of diabetes mellitus. AB - BACKGROUND: Multiple laboratory tests are used to diagnose and manage patients with diabetes mellitus. The quality of the scientific evidence supporting the use of these tests varies substantially. APPROACH: An expert committee compiled evidence-based recommendations for the use of laboratory testing for patients with diabetes. A new system was developed to grade the overall quality of the evidence and the strength of the recommendations. Draft guidelines were posted on the Internet and presented at the 2007 Arnold O. Beckman Conference. The document was modified in response to oral and written comments, and a revised draft was posted in 2010 and again modified in response to written comments. The National Academy of Clinical Biochemistry and the Evidence Based Laboratory Medicine Committee of the AACC jointly reviewed the guidelines, which were accepted after revisions by the Professional Practice Committee and subsequently approved by the Executive Committee of the American Diabetes Association. CONTENT: In addition to long-standing criteria based on measurement of plasma glucose, diabetes can be diagnosed by demonstrating increased blood hemoglobin A(1c) (Hb A(1c)) concentrations. Monitoring of glycemic control is performed by self-monitoring of plasma or blood glucose with meters and by laboratory analysis of Hb A(1c). The potential roles of noninvasive glucose monitoring, genetic testing, and measurement of autoantibodies, urine albumin, insulin, proinsulin, C-peptide, and other analytes are addressed. SUMMARY: The guidelines provide specific recommendations that are based on published data or derived from expert consensus. Several analytes have minimal clinical value at present, and their measurement is not recommended. PMID- 21617153 TI - Executive summary: guidelines and recommendations for laboratory analysis in the diagnosis and management of diabetes mellitus. AB - BACKGROUND: Multiple laboratory tests are used in the diagnosis and management of patients with diabetes mellitus. The quality of the scientific evidence supporting the use of these assays varies substantially. APPROACH: An expert committee compiled evidence-based recommendations for the use of laboratory analysis in patients with diabetes. A new system was developed to grade the overall quality of the evidence and the strength of the recommendations. A draft of the guidelines was posted on the Internet, and the document was modified in response to comments. The guidelines were reviewed by the joint Evidence-Based Laboratory Medicine Committee of the AACC and the National Academy of Clinical Biochemistry and were accepted after revisions by the Professional Practice Committee and subsequent approval by the Executive Committee of the American Diabetes Association. CONTENT: In addition to the long-standing criteria based on measurement of venous plasma glucose, diabetes can be diagnosed by demonstrating increased hemoglobin A(1c) (Hb A(1c)) concentrations in the blood. Monitoring of glycemic control is performed by the patients measuring their own plasma or blood glucose with meters and by laboratory analysis of Hb A(1c). The potential roles of noninvasive glucose monitoring, genetic testing, and measurement of autoantibodies, urine albumin, insulin, proinsulin, C-peptide, and other analytes are addressed. SUMMARY: The guidelines provide specific recommendations based on published data or derived from expert consensus. Several analytes are found to have minimal clinical value at the present time, and measurement of them is not recommended. PMID- 21617156 TI - Correlations among various functional and morphological tests in resolved central serous chorioretinopathy. AB - AIMS: To find the explanatory parameters for best corrected visual acuity (BCVA) and microperimetry (MP) in resolved central serous chorioretinopathy. METHODS: Thirty-three eyes from 33 patients were analysed retrospectively. BCVA and MP were correlated with parameters from various functional and morphological tests. The mean MP sensitivities at fovea 2 degrees and 4 degrees , retinal thickness and degree of defect at the photoreceptor inner and outer segment junction (IS/OS) of the spectral domain-optical coherent tomography image, normalised coefficient of variation of grey scale at the fovea in the short wavelength and near infrared fundus autofluorescence image, P1 amplitude and latency, and N1 amplitude and latency of multifocal electroretinography at ring 1 (R1) were measured. Spearman correlations and multiple linear regression analysis were used for analysis of correlation between functional and morphological characteristics. RESULTS: The mean BCVA was 0.19 +/- 0.23 (logarithm of the minimum angle of resolution (logMAR)). The mean MP at 2 degrees was 12.79 +/- 4.47 dB. BCVA correlated with MP at 2 degrees (rho=-0.491, p=0.004) and had a significant association with the IS/OS defect and centre point thickness (CPT) (BCVA=0.413+0.314 * IS/OS defect-0.002 * CPT; p<0.001, R=0.739, R(2)=0.546). Retinal sensitivity measured by MP at the fovea (2 degrees ) had a significant association with the IS/OS defect and N1 latency at R1 (MP at 2 degrees =19.350 9.116 * IS/OS defect -0.324 * N1 latency at R1; p<0.001, R=0.804, R(2)=0.647). CONCLUSIONS: The visual function of eyes with resolved central serous chorioretinopathy was suboptimal and was better explained when various parameters showing retinal status were combined and interpreted together. PMID- 21617157 TI - 24-hour ocular perfusion pressure in glaucoma patients. PMID- 21617155 TI - Discovery and assessment of conserved Pax6 target genes and enhancers. AB - The characterization of transcriptional networks (TNs) is essential for understanding complex biological phenomena such as development, disease, and evolution. In this study, we have designed and implemented a procedure that combines in silico target screens with zebrafish and mouse validation, in order to identify cis-elements and genes directly regulated by Pax6. We chose Pax6 as the paradigm because of its crucial roles in organogenesis and human disease. We identified over 600 putative Pax6 binding sites and more than 200 predicted direct target genes, conserved in evolution from zebrafish to human and to mouse. This was accomplished using hidden Markov models (HMMs) generated from experimentally validated Pax6 binding sites. A small sample of genes, expressed in the neural lineage, was chosen from the predictions for RNA in situ validation using zebrafish and mouse models. Validation of DNA binding to some predicted cis elements was also carried out using chromatin immunoprecipitation (ChIP) and zebrafish reporter transgenic studies. The results show that this combined procedure is a highly efficient tool to investigate the architecture of TNs and constitutes a useful complementary resource to ChIP and expression data sets because of its inherent spatiotemporal independence. We have identified several novel direct targets, including some putative disease genes, among them Foxp2; these will allow further dissection of Pax6 function in development and disease. PMID- 21617158 TI - Allogeneic mesenchymal stem cell transplantation in seven patients with refractory inflammatory bowel disease. PMID- 21617154 TI - Photodynamic therapy of cancer: an update. AB - Photodynamic therapy (PDT) is a clinically approved, minimally invasive therapeutic procedure that can exert a selective cytotoxic activity toward malignant cells. The procedure involves administration of a photosensitizing agent followed by irradiation at a wavelength corresponding to an absorbance band of the sensitizer. In the presence of oxygen, a series of events lead to direct tumor cell death, damage to the microvasculature, and induction of a local inflammatory reaction. Clinical studies revealed that PDT can be curative, particularly in early stage tumors. It can prolong survival in patients with inoperable cancers and significantly improve quality of life. Minimal normal tissue toxicity, negligible systemic effects, greatly reduced long-term morbidity, lack of intrinsic or acquired resistance mechanisms, and excellent cosmetic as well as organ function-sparing effects of this treatment make it a valuable therapeutic option for combination treatments. With a number of recent technological improvements, PDT has the potential to become integrated into the mainstream of cancer treatment. PMID- 21617160 TI - Impressive hypertensive encephalopathy. PMID- 21617159 TI - Interhospital variation in the RATPAC trial (Randomised Assessment of Treatment using Panel Assay of Cardiac markers). AB - BACKGROUND: The RATPAC trial showed that using a point-of-care panel of CK MB(mass), myoglobin and troponin at baseline and 90 min increased the proportion of patients successfully discharged home, leading to reduced median length of initial hospital stay. However, it did not change mean hospital stay and may have increased mean costs per patient. The aim of this study was to explore variation in outcome and costs between participating hospitals. METHODS: RATPAC was a pragmatic multicentre randomised controlled trial (N=2243) and economic analysis comparing diagnostic assessment using the panel to standard care for patients with acute chest pain due to suspected myocardial infarction at six hospitals. The difference in the proportion of patients successfully discharged (primary outcome) and mean costs per patient between the participating hospitals was compared. RESULTS: Point-of-care assessment led to a higher proportion of successful discharges in four hospitals, a lower proportion in one and was equivocal in another. The OR (95% CI) for the primary outcome varied from 0.12 (0.01 to 1.03) to 11.07 (6.23 to 19.66) with significant heterogeneity between the centres (p<0.001). The mean cost per patient for the intervention group ranged from being L214.49 less than the control group (-132.56 to 657.10) to L646.57 more expensive (73.12 to 1612.71), with weak evidence of heterogeneity between the centres (p=0.0803). CONCLUSION: The effect of point-of-care panel assessment on successful discharge and costs per patient varied markedly between hospitals and may depend on local protocols, staff practices and available facilities. PMID- 21617161 TI - A cohort study of outcomes following head injury among children and young adults in full-time education. AB - OBJECTIVE: To estimate the prevalence of post-concussive symptoms (PCS) following head injury among adolescents in full-time education and to identify prognostic factors at presentation to the emergency department (ED) that may predict the development of PCS. METHODS: An observational cohort study of all head injured patients aged 13-21 and in full-time education presenting to an inner city ED was performed. Subjects were followed up at 1 and 6 months after injury by structured telephone interview to assess for the presence of symptoms or ongoing disability. Presentation data of those identified as having PCS underwent regression analysis to isolate potential prognostic indicators for such problems. RESULTS: Of the 188 patients recruited, 5.9% (95% CI 3.3% to 10.2%) still had some symptoms after 6 months, with half of these claiming that such symptoms were affecting everyday living. Of these patients, 82% were assaulted as the cause of their injury and nearly 40% had no conventional indicators of head injury severity at presentation. After 1 month, 46/188 (24.5%, 95% CI 18.9% to 31.1%) patients had some degree of symptoms, most of whom were discharged directly from the ED. Potential prognostic indicators identified were a reduced Glasgow Coma Score (GCS) (<15) at presentation and being assaulted as the cause of injury. CONCLUSION: The prevalence of PCS 6 months following head injury for the selected sub-group was 5.9%, and 10.6% if assaulted. Most patients who developed PCS were discharged directly from the ED. PMID- 21617162 TI - Genetics and cardiovascular disease--causes and prevention of unexpected sudden adult death: the role of the SADS clinic. AB - Sudden arrhythmic death syndrome (SADS) accounts for approximately 500 deaths in England and Wales per year. Clinical screening of the surviving first-degree relatives can identify an inherited cardiovascular condition in up to half of families, permitting lifestyle modification and confirmed effective prophylactic therapies to prevent further sudden deaths. Mechanisms for molecular autopsy are available to improve the diagnostic yield but practical barriers to its successful implementation exist. This article reviews the clinical screening of the first-degree relatives of SADS patients, molecular autopsy of probands and the broader implications of national recommendations for the investigation of sudden cardiac death. PMID- 21617163 TI - Prospective randomised trial evaluating a paclitaxel-coated balloon in patients treated with endothelial progenitor cell capturing stents for de novo coronary artery disease. AB - BACKGROUND: Percutaneous coronary intervention with stent implantation is limited by the occurrence of re-stenosis and the risk of stent thromboses. OBJECTIVE: To define the impact of paclitaxel-coated balloon angioplasty plus endothelial progenitor cell capturing (EPC) stent implantation in de novo coronary artery disease. This combination may reduce neointimal proliferation within the EPC stent and address the risk of stent thrombosis by facilitating rapid endothelialisation. METHODS: In this prospective single-blind multicentre randomised trial, 120 patients with a de novo lesion in a native coronary artery were randomly assigned to undergo treatment with paclitaxel-coated balloon plus EPC stent or EPC stent alone. Dual antiplatelet therapy was prescribed for 3 months. Angiographic follow-up was scheduled at 6 months. The primary endpoint was in-stent late lumen loss. The secondary clinical endpoint was a composite of death from a cardiac cause, myocardial infarction attributed to the target vessel or target lesion revascularisation. RESULTS: There was no difference in patient baseline characteristics or procedural results. The angiographic follow-up rate was 96%. Treatment with paclitaxel-coated balloon plus EPC stent was superior to EPC stent alone, with an in-stent late loss of 0.34+/-0.45 mm versus 0.88+/-0.48 mm (p<0.001). The re-stenosis rate was reduced from 23.2% to 5.1% (p=0.006) and the clinical endpoint was reduced from 17.2% to 4.8% (p=0.039). There was no definite or probable stent thrombosis. CONCLUSIONS: Paclitaxel-coated balloon plus EPC stent implantation is superior to EPC stent implantation alone for treatment of de novo coronary artery disease. TRIAL REGISTRATION: NCT00732953. PMID- 21617164 TI - From genomic databases to translation: a call to action. AB - The rapid rise of international collaborative science has enabled access to genomic data. In this article, it is argued that to move beyond mapping genomic variation to understanding its role in complex disease aetiology and treatment will require extending data sharing for the purposes of clinical research translation and implementation. PMID- 21617165 TI - The morality of care: case study and review. AB - This case concerns aspects of the treatment of a post-surgical patient in a major public hospital in New Zealand during the author's experiences as a fourth year medical student. This case is used to consider the interlinked ethical issues of sympathy, moral virtue, dignity and how the medical environment can realign these values. PMID- 21617166 TI - What context features might be important determinants of the effectiveness of patient safety practice interventions? AB - BACKGROUND: Differences in contexts (eg, policies, healthcare organisation characteristics) may explain variations in the effects of patient safety practice (PSP) implementations. However, knowledge of which contextual features are important determinants of PSP effectiveness is limited and consensus is lacking on a taxonomy of which contexts matter. METHODS: Iterative, formal discussions were held with a 22-member technical expert panel composed of experts or leaders in patient safety, healthcare systems, and methods. First, potentially important contextual features were identified, focusing on five PSPs. Then, two surveys were conducted to determine the context likely to influence PSP implementations. RESULTS: The panel reached a consensus on a taxonomy of four broad domains of contextual features important for PSP implementations: safety culture, teamwork and leadership involvement; structural organisational characteristics (eg, size, organisational complexity or financial status); external factors (eg, financial or performance incentives or PSP regulations); and availability of implementation and management tools (eg, training organisational incentives). Panelists also tended to rate specific patient safety culture, teamwork and leadership contexts as high priority for assessing their effects on PSP implementations, but tended to rate specific organisational characteristic contexts as high priority only for use in PSP evaluations. Panelists appeared split on whether specific external factors and implementation/management tools were important for assessment or only description. CONCLUSION: This work can guide research commissioners and evaluators on the contextual features of PSP implementations that are important to report or evaluate. It represents a first step towards developing guidelines on contexts in PSP implementation evaluations. However, the science of context measurement needs maturing. PMID- 21617167 TI - Prospective comparison of three guideline development methods for treatment of actinic keratosis. AB - OBJECTIVE: To compare three methods of guideline development, to see whether using alternative evidence-based methods resulted in variation of recommendations for treating actinic keratosis. METHODS: Method 1 followed a standard multiple session evidence-based approach with a working group. In method 2 recommendations were formulated by a working group during a 2-day conference. Method 3 used one epidemiologist to summarise the evidence and one dermatologist to make clinical recommendations afterwards. Graded recommendations and levels of evidence were compared per therapy across three draft guidelines. The primary outcome was the extent of accordance or discordance. Secondary outcomes were total costs and time period necessary to make a draft guideline. RESULTS: Therapeutic recommendations and levels of evidence differed in some occasions. However, intraclass correlations between levels of evidence were significant (method 1 vs 2: p = 0.003; method 1 vs 3: p < 0.001). Regarding recommendation variation method 1 and method 2 correlated significant at 0.755 (p = 0.001). Method 1 versus 3 and method 2 versus 3 also showed significant, but lower, correlation coefficients (respectively, 0.493 (p = 0.026) and 0.673 (p = 0.007)). Method 3 was the cheapest and quickest (24,770 euro and 4 months) and method 1 was the most expensive and slowest method (?48,100 euro and 14 months). CONCLUSIONS: The value of a guideline using alternative evidence-based methods seems to at least equal that of a guideline composed in multiple sessions, that is, for topics with a monodisciplinary character and a relatively small number of conducted trials. In addition, the presented alternatives were more time- and cost-efficient. PMID- 21617168 TI - The fibrinogen cleavage product Aalpha-Val360, a specific marker of neutrophil elastase activity in vivo. AB - BACKGROUND: Alpha-1-antitrypsin (A1AT) deficiency is the only recognised genetic risk factor for chronic obstructive pulmonary disease (COPD), a leading cause of morbidity and mortality worldwide. Since A1AT is the major inhibitor of neutrophil elastase (NE), this enzyme has become widely implicated in the pathogenesis of COPD in general; however, there is currently no specific biomarker for its pre-inhibition activity. Such a biomarker should be a measure of elastase-specific COPD disease activity with the potential to assess early targeted therapeutic intervention, in contrast to traditional and non-specific disease severity markers such as forced expiratory volume in 1 s. METHODS: In pilot studies, plasma Aalpha-Val(360) and markers of neutrophil activation were measured in 95 subjects with a range of A1AT concentrations. Aalpha-Val(360) and sputum elastase activity were also measured in a further seven PiZ A1AT-deficient subjects over the course of an acute exacerbation. Finally, Aalpha-Val(360) was measured in plasma from subjects randomised to receive A1AT replacement or placebo in the EXACTLE trial. RESULTS: The plasma concentrations of Aalpha Val(360) and A1AT related exponentially, consistent with previous theoretical and in vitro experimental data. L-233 (an intracellular NE inhibitor) blocked generation of Aalpha-Val(360) and subsequent A1AT/NE complex formation. Aalpha Val(360) was related to the spirometric severity of lung disease in A1AT deficiency, to sputum elastase activity in acute exacerbations and was decreased in subjects receiving A1AT replacement therapy (while remaining constant in those receiving placebo). CONCLUSIONS: Aalpha-Val(360) represents the first specific footprint of pre-inhibition NE activity and is a potential biomarker of disease activity and progression in subjects with elastase-dependent COPD. TRIAL REGISTRATION: The EXACTLE study was registered in ClinicalTrials.gov as 'Antitrypsin (AAT) to Treat Emphysema in AAT-Deficient Patients'; ClinicalTrials.gov Identifier: NCT00263887. PMID- 21617170 TI - A spline for the time. PMID- 21617169 TI - Emergence of parapneumonic empyema in the USA. AB - BACKGROUND: Although recent reports suggest that the incidence of parapneumonic empyema has increased in several regions of the USA, national trends in disease burden are unknown. National trends in the incidence of parapneumonic empyema hospitalisations and changes in empyema by associated pathogens were examined. METHODS: National hospitalisation data (1996-2008) were analysed and rates estimated using census estimates as denominators. Incidence rate ratios (IRR) compared 2008 with 1996 rates. Discharge diagnosis codes were used to characterise pathogens associated with empyema hospitalisations. RESULTS: Overall, national parapneumonic empyema-related hospitalisation rates increased from 3.04 per 100,000 in 1996 to 5.98 per 100,000 in 2008, a 2.0-fold increase (95% CI 1.8 to 2.1). The increases were observed among children (IRR 1.9 (95% CI 1.4 to 2.7)) and adults aged 18-39, 40-64 and >=65 years (IRR 1.8 (95% CI 1.5 to 2.1), 2.0 (95% CI 1.6 to 3.1) and 1.7 (95% CI 1.5 to 2.0), respectively). Overall, pneumococcal empyema rates remained relatively stable in all age groups whereas streptococcal- (non-pneumococcal) and staphylococcal-related empyema rates increased 1.9-fold and 3.3-fold, respectively, with consistent increases across age groups. The overall in-hospital case fatality ratio for parapneumonic empyema-related hospitalisations was 8.0% (95% CI 6.4% to 9.5%) in 1996 and 7.2% (95% CI 6.3% to 8.1%) in 2008 (p=0.395). Of the empyemas where study pathogens were listed (37.6%), staphylococcal-related empyema had the largest absolute increases across age groups and was associated with longer hospital stay and higher in-hospital mortality than other empyemas. CONCLUSIONS: Although parapneumonic empyema-related hospitalisations remained relatively rare, they increased substantially during the study period. A number of pathogens, especially staphylococcus, contributed to this increase. PMID- 21617171 TI - PCR-based tissue identification: the UCLH experience. AB - The need to accurately identify tissue of an individual can arise in a variety of settings including mislabelled slides or sample carryover. Reported rates of carryover range from 0.6% to 2.9% of slides depending on the methods of evaluation. Carryover becomes particularity clinically important when malignant tissue is found in an otherwise benign sample. The suspicion of malignancy causes immense psychological stress to the patient and results in additional management costs due to the additional investigations required to rule out malignancy. Proving a negative can be difficult and many cases result in lifelong follow-up for the patient. Molecular techniques such as PCR amplification of simple tandem repeat (STR) sequences can be used to identify tissue and hence its provenance. At University College London Hospital, STR PCR analysis has been used since 2003. Here the authors report their experience with regard to the clinical scenarios, the technique used and the outcomes. PMID- 21617172 TI - Bilateral thalamic haemorrhage and intracranial injuries related to alcohol induced thrombocytopenia. PMID- 21617173 TI - The criminal justice system's considerations of so-called near-virtual autopsies: the East Midlands experience. AB - BACKGROUND: While several research groups champion the potential for postmortem CT (PMCT) to replace the invasive postmortem (PM), many questions still remain. AIMS: Perhaps the two most important questions are whether PMCT can provide the same level of information as an invasive PM, and arguably more importantly, can it meet the needs of the end users of the PM report. Through a comparative analysis of invasive post-mortem and CT findings and a questionnaire based qualitative thematic analysis, the authors have sought to answer these questions. RESULTS AND CONCLUSION: Here, the authors show that PMCT is good at providing accurate causes of death and that the interpretation of cases is not significantly altered by the absence of histology. The authors show that in straightforward trauma deaths such as road traffic incidents, there exists the potential for the replacement of the invasive PM by PMCT examination. However, as yet, PMCT cannot provide all of the information that is expected by the criminal justice system in complex forensic cases. PMID- 21617175 TI - Republished paper: assessing and improving safety culture throughout an academic medical centre: a prospective cohort study. AB - OBJECTIVES To describe the authors' hospital-wide efforts to improve safety climate at a large academic medical centre. DESIGN AND SETTING A prospective cohort study used multiple interventions to improve hospital-wide safety climate. 144 clinical units in an urban academic medical centre are included in this analysis. Interventions The comprehensive unit-based safety programme included steps to identify hazards, partner units with a senior executive to fix hazards, learn from defects, and implement communication and teamwork tools. Hospital level interventions were also implemented. Main outcome measures Safety climate was assessed annually using the safety attitudes questionnaire. The safety culture goal was to meet or exceed the 60% minimum positive score or improve the score by >=10 points. RESULTS Response rates were 77% (2006) and 79% (2008). For safety climate, 55% of units in 2006 and 82% in 2008 achieved the culture goal. For teamwork climate, 61% of units in 2006 and 83% in 2008 achieved the culture goal. The mean safety climate improvement (difference score) for 79 units at or above 60% in 2006 was 0.201 in 2008; the mean improvement for the 65 units below the threshold was 18.278. The mean teamwork climate improvement (difference score) for the 89 units at or above 60% in 2006 was 0.452 in 2008; the mean improvement for the 55 units below the threshold was 16.176. Climate scores improved significantly from 2006 to 2008 in every domain except stress recognition. CONCLUSIONS Hospital-wide interventions were associated with improvements in safety climate at a large academic medical centre. PMID- 21617174 TI - Global DNA hypomethylation is an early event in Helicobacter pylori-related gastric carcinogenesis. AB - AIM: Cancer, particularly gastric cancer (GC), is prevalently an epigenetic phenomenon that is dependent on an altered DNA methylation pattern. In gastric carcinogenesis, many genes show aberrant methylation; however, none of them may be used as a biomarker of cancer risk and progression. The authors aimed to evaluate the global DNA methylation of gastric mucosa in Helicobacter pylori (Hp) related chronic gastritis, in GC and in 10 patients with preneoplastic lesions (ie, atrophy and intestinal metaplasia) followed up for 10 years. METHODS: The authors analysed 93 dyspeptic patients who underwent upper endoscopy, 41 surgical GC samples and 10 patients with preneoplastic gastric lesions followed up for 10 years after successful Hp eradication therapy. Global DNA methylation status and surrogate markers of cell proliferation and apoptosis were evaluated by immunohistochemistry using the anti-5-methylcytosine (5-MC), anti-Ki-67 and anti p53 (anti-apoptotic marker)-specific antibodies, respectively. RESULTS: Global DNA methylation of gastric mucosa gradually decreased from normal mucosa to Hp positive gastritis, Hp-positive chronic atrophic gastritis, independent of Cag-A status and GC; however, the variation was significant (p<0.05) only between Hp negative subjects and Hp-positive chronic gastritis. Interestingly, the 5-MC immunostaining was absent in areas of intestinal metaplasia. In the 10 patients with preneoplastic lesions, global DNA methylation decreased over time despite the eradication of Hp infection, but reached significance only at 10 years versus baseline. The 5-MC immunostaining negatively correlated with Ki-67 and p53 expression in all groups. CONCLUSION: Global DNA hypomethylation is an early molecular event in Hp-related gastric carcinogenesis. Further studies with more cases and a longer follow-up are needed to establish the potential GC predictive role of DNA hypomethylation. PMID- 21617176 TI - Identifying children at risk for type 2 diabetes in underserved communities. AB - PURPOSE: The purpose of this study was to identify and assess health behaviors among ethnic minority children at high risk for type 2 diabetes. METHODS: Diabetes screenings were conducted at community centers, churches, and local neighborhood health fairs in Madison, Wisconsin. During these events, diabetes risk assessment surveys were given to parents of children between the ages of 10 to 19 years. Parents who identified their children as having 2 or more risk factors for type 2 diabetes were invited to have their child screened for type 2 diabetes. RESULTS: A total of 86 children between the ages of 10 to 19 years (mean age = 13; 58% male) were screened for diabetes. Fifty-one percent of the children were overweight or obese with 38% having >3 risk factors for type 2 diabetes. While there was no significant difference in the nutritional habits reported between normal, overweight, or obese children, fewer overweight and obese children reported exercising at least 30 minutes 5 to 7 days a week compared to children with a normal weight (P = .033). CONCLUSION: Prevention of diabetes is a powerful public health intervention. Targeted diabetes screening in disadvantaged, underserved communities is an effective way to identify families with children at risk for type 2 diabetes. In addition, information obtained from these screenings can assist researchers and clinicians in designing accessible and affordable health promotion interventions that are culturally relevant to the youth and families within the community. PMID- 21617177 TI - Thrombospondin-1: an islet endothelial cell signal of importance for beta-cell function. AB - OBJECTIVE: Loss of thrombospondin (TSP)-1 in pancreatic islets has been shown to cause islet hyperplasia. This study tested the hypothesis that endothelial derived TSP-1 is important for beta-cell function. RESEARCH DESIGN AND METHODS: Islet function was evaluated both in vivo and in vitro. Messenger RNA and protein expression were measured by real-time PCR and Western blot, respectively. The role of endothelial-derived TSP-1 for beta-cell function was determined using a transplantation design in which recipient blood vessels either were allowed to grow or not into the transplanted islets. RESULTS: TSP-1-deficient mice were glucose intolerant, despite having an increased beta-cell mass. Moreover, their islets had decreased glucose-stimulated insulin release, (pro)insulin biosynthesis, and glucose oxidation rate, as well as increased expression of uncoupling protein-2 and lactate dehydrogenase-A when compared with control islets. Almost all TSP-1 in normal islets were found to be derived from the endothelium. Transplantation of free and encapsulated neonatal wild-type and TSP 1-deficient islets was performed in order to selectively reconstitute with TSP-1 positive or -negative blood vessels in the islets and supported that the beta cell defects occurring in TSP-1-deficient islets reflected postnatal loss of the glycoprotein in the islet endothelial cells. Treatment of neonatal TSP-1 deficient mice with the transforming growth factor (TGF)beta-1-activating sequence of TSP-1 showed that reconstitution of TGFbeta-1 activation prevented the development of decreased glucose tolerance in these mice. Thus, endothelial derived TSP-1 activates islet TGFbeta-1 of importance for beta-cells. CONCLUSIONS: Our study indicates a novel role for endothelial cells as functional paracrine support for pancreatic beta-cells. PMID- 21617178 TI - Transcriptional profiling of diabetic neuropathy in the BKS db/db mouse: a model of type 2 diabetes. AB - OBJECTIVE: A better understanding of the molecular mechanisms underlying the development and progression of diabetic neuropathy (DN) is essential for the design of mechanism-based therapies. We examined changes in global gene expression to define pathways regulated by diabetes in peripheral nerve. RESEARCH DESIGN AND METHODS: Microarray data for 24-week-old BKS db/db and db/+ mouse sciatic nerve were analyzed to define significantly differentially expressed genes (DEGs); DEGs were further analyzed to identify regulated biological processes and pathways. Expression profile clustering was performed to identify coexpressed DEGs. A set of coexpressed lipid metabolism genes was used for promoter sequence analysis. RESULTS: Gene expression changes are consistent with structural changes of axonal degeneration. Pathways regulated in the db/db nerve include lipid metabolism, carbohydrate metabolism, energy metabolism, peroxisome proliferator-activated receptor signaling, apoptosis, and axon guidance. Promoter sequences of lipid metabolism-related genes exhibit evidence of coregulation of lipid metabolism and nervous system development genes. CONCLUSIONS: Our data support existing hypotheses regarding hyperglycemia-mediated nerve damage in DN. Moreover, our analyses revealed a possible coregulation mechanism connecting hyperlipidemia and axonal degeneration. PMID- 21617180 TI - Genetic defect in phospholipase Cdelta1 protects mice from obesity by regulating thermogenesis and adipogenesis. AB - OBJECTIVE: Regulation of obesity development is an important issue to prevent metabolic syndromes. Gene-disrupted mice of phospholipase Cdelta1 (PLCdelta1), a key enzyme of phosphoinositide turnover, seemed to show leanness. Here we examined whether and how PLCdelta1 is involved in obesity. RESEARCH DESIGN AND METHODS: Weight gain, insulin sensitivity, and metabolic rate in PLCdelta1(-/-) mice were compared with PLCdelta1(+/-) littermate mice on a high-fat diet. Thermogenic and adipogenetic potentials of PLCdelta1(-/-) immortalized brown adipocytes and adipogenesis of PLCdelta1-knockdown (KD) 3T3L1 cells, or PLCdelta1(-/-) white adipose tissue (WAT) stromal-vascular fraction (SVF) cells, were also investigated. RESULTS: PLCdelta1(-/-) mice showed marked decreases in weight gain and mass of epididymal WAT and preserved insulin sensitivity compared with PLCdelta1(+/-) mice on a high-fat diet. In addition, PLCdelta1(-/-) mice have a higher metabolic rate such as higher oxygen consumption and heat production. When control immortalized brown adipocytes were treated with thermogenic inducers, expression of PLCdelta1 was decreased and thermogenic gene uncoupling protein 1 (UCP1) was upregulated to a greater extent in PLCdelta1(-/-) immortalized brown adipocytes. In contrast, ectopic expression of PLCdelta1 in PLCdelta1(-/-) brown adipocytes induced a decrease in UCP expression, indicating that PLCdelta1 negatively regulates thermogenesis. Importantly, accumulation of lipid droplets was severely decreased when PLCdelta1-KD 3T3L1 cells, or PLCdelta1(-/-) WAT SVF cells, were differentiated, whereas differentiation of PLCdelta1(-/-) brown preadipocytes was promoted. CONCLUSIONS: PLCdelta1 has essential roles in thermogenesis and adipogenesis and thereby contributes to the development of obesity. PMID- 21617179 TI - Body weight, not insulin sensitivity or secretion, may predict spontaneous weight changes in nondiabetic and prediabetic subjects: the RISC study. AB - OBJECTIVE: Previous studies have found that high insulin sensitivity predicts weight gain; this association has not been confirmed. Our aim was to systematically analyze metabolic predictors of spontaneous weight changes. RESEARCH DESIGN AND METHODS: In 561 women and 467 men from the Relationship Between Insulin Sensitivity and Cardiovascular Disease (RISC) cohort (mean age 44 years, BMI range 19-44 kg/m(2), 9% impaired glucose tolerance) followed up for 3 years, we measured insulin sensitivity (by a euglycemic clamp) and beta-cell function (by modeling of the C-peptide response to oral glucose and by acute insulin response to intravenous glucose). RESULTS: Insulin sensitivity was similar in weight gainers (top 20% of the distribution of BMI changes), weight losers (bottom 20%), and weight stable subjects across quartiles of baseline BMI. By multiple logistic or linear regression analyses controlling for center, age, sex, and baseline BMI, neither insulin sensitivity nor any beta-cell function parameter showed an independent association with weight gain; this was true in normal glucose tolerance, impaired glucose tolerance, and whether subjects progressed to dysglycemia or not. Baseline BMI was significantly higher in gainers (26.1 +/- 4.1 kg/m(2)) and losers (26.6 +/- 3.7 kg/m(2)) than in weight stable subjects (24.8 +/- 3.8 kg/m(2), P<0.0001 for both gainers and losers). Baseline waist circumference (or equivalently, BMI or weight) was a positive, independent predictor of both weight gain and weight loss (odds ratio 1.48 [95% CI 1.12-1.97]) in men and (1.67 [1.28-2.12]) in women. In men only, better insulin sensitivity was an additional independent predictor of weight loss. CONCLUSIONS: Neither insulin sensitivity nor insulin secretion predicts spontaneous weight gain. Individuals who have attained a higher weight are prone to either gaining or losing weight regardless of their glucose tolerance. PMID- 21617181 TI - Activation of peroxisome proliferator-activated receptor-beta/-delta (PPAR-beta/ delta) ameliorates insulin signaling and reduces SOCS3 levels by inhibiting STAT3 in interleukin-6-stimulated adipocytes. AB - OBJECTIVE: It has been suggested that interleukin (IL)-6 is one of the mediators linking obesity-derived chronic inflammation with insulin resistance through activation of STAT3, with subsequent upregulation of suppressor of cytokine signaling 3 (SOCS3). We evaluated whether peroxisome proliferator-activated receptor (PPAR)-beta/-delta prevented activation of the IL-6-STAT3-SOCS3 pathway and insulin resistance in adipocytes. RESEARCH DESIGN AND METHODS: Adipocytes and white adipose tissue from wild-type and PPAR-beta/-delta-null mice were used to evaluate the effect of PPAR-beta/-delta on the IL-6-STAT3-SOCS3 pathway. RESULTS: First, we observed that the PPAR-beta/-delta agonist GW501516 prevented both IL-6 dependent reduction in insulin-stimulated Akt phosphorylation and glucose uptake in adipocytes. In addition, this drug treatment abolished IL-6-induced SOCS3 expression in differentiated 3T3-L1 adipocytes. This effect was associated with the capacity of the drug to prevent IL-6-induced STAT3 phosphorylation on Tyr(705) and Ser(727) residues in vitro and in vivo. Moreover, GW501516 prevented IL-6-dependent induction of extracellular signal-related kinase (ERK)1/2, a serine-threonine-protein kinase involved in serine STAT3 phosphorylation. Furthermore, in white adipose tissue from PPAR-beta/-delta-null mice, STAT3 phosphorylation (Tyr(705) and Ser(727)), STAT3 DNA-binding activity, and SOCS3 protein levels were higher than in wild-type mice. Several steps in STAT3 activation require its association with heat shock protein 90 (Hsp90), which was prevented by GW501516 as revealed in immunoprecipitation studies. Consistent with this finding, the STAT3-Hsp90 association was enhanced in white adipose tissue from PPAR-beta/-delta-null mice compared with wild-type mice. CONCLUSIONS: Collectively, our findings indicate that PPAR-beta/-delta activation prevents IL 6-induced STAT3 activation by inhibiting ERK1/2 and preventing the STAT3-Hsp90 association, an effect that may contribute to the prevention of cytokine-induced insulin resistance in adipocytes. PMID- 21617182 TI - Glycation of LDL by methylglyoxal increases arterial atherogenicity: a possible contributor to increased risk of cardiovascular disease in diabetes. AB - OBJECTIVE: To study whether modification of LDL by methylglyoxal (MG), a potent arginine-directed glycating agent that is increased in diabetes, is associated with increased atherogenicity. RESEARCH DESIGN AND METHODS: Human LDL was isolated and modified by MG in vitro to minimal extent (MG(min)-LDL) as occurs in vivo. Atherogenic characteristics of MG(min)-LDL were characterized: particle size, proteoglycan-binding, susceptibility to aggregation, LDL and non-LDL receptor-binding, and aortal deposition. The major site of modification of apolipoprotein B100 (apoB100) modification was investigated by mass spectrometric peptide mapping. RESULTS: MG(min)-LDL contained 1.6 molar equivalents of MG modification-mostly hydroimidazolone-as found in vivo. MG(min)-LDL had decreased particle size, increased binding to proteoglycans, and increased aggregation in vitro. Cell culture studies showed that MG(min)-LDL was bound by the LDL receptor but not by the scavenger receptor and had increased binding affinity for cell surface heparan sulfate-containing proteoglycan. Radiotracer studies in rats showed that MG(min)-LDL had a similar fractional clearance rate in plasma to unmodified LDL but increased partitioning onto the aortal wall. Mass spectrometry peptide mapping identified arginine-18 as the hotspot site of apoB100 modification in MG(min)-LDL. A computed structural model predicted that MG modification of apoB100 induces distortion, increasing exposure of the N-terminal proteoglycan-binding domain on the surface of LDL. This likely mediates particle remodeling and increases proteoglycan binding. CONCLUSIONS: MG modification of LDL forms small, dense LDL with increased atherogenicity that provides a new route to atherogenic LDL and may explain the escalation of cardiovascular risk in diabetes and the cardioprotective effect of metformin. PMID- 21617183 TI - Comment on: Atkinson. It's time to consider changing the rules: the rationale for rethinking control groups in clinical trials aimed at reversing type 1 diabetes. Diabetes 2011;60:361-363. PMID- 21617186 TI - Stimulating IL-13 receptors on T cells: a new pathway for tolerance induction in diabetes? PMID- 21617185 TI - Efficacy of metreleptin in obese patients with type 2 diabetes: cellular and molecular pathways underlying leptin tolerance. AB - OBJECTIVE: Metreleptin has been efficacious in improving metabolic control in patients with lipodystrophy, but its efficacy has not been tested in obese patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: We studied the role of leptin in regulating the endocrine adaptation to long-term caloric deprivation and weight loss in obese diabetic subjects over 16 weeks in the context of a double-blinded, placebo-controlled, randomized trial. We then performed detailed interventional and mechanistic signaling studies in humans in vivo, ex vivo, and in vitro. RESULTS: In obese patients with diabetes, metreleptin administration for 16 weeks did not alter body weight or circulating inflammatory markers but reduced HbA(1c) marginally (8.01 +/- 0.93-7.96 +/- 1.12, P = 0.03). Total leptin, leptin-binding protein, and antileptin antibody levels increased, limiting free leptin availability and resulting in circulating free leptin levels of ~50 ng/mL. Consistent with clinical observations, all metreleptin signaling pathways studied in human adipose tissue and peripheral blood mononuclear cells were saturable at ~50 ng/mL, with no major differences in timing or magnitude of leptin-activated STAT3 phosphorylation in tissues from male versus female or obese versus lean humans in vivo, ex vivo, or in vitro. We also observed for the first time that endoplasmic reticulum (ER) stress in human primary adipocytes inhibits leptin signaling. CONCLUSIONS: In obese patients with diabetes, metreleptin administration did not alter body weight or circulating inflammatory markers but reduced HbA(1c) marginally. ER stress and the saturable nature of leptin signaling pathways play a key role in the development of leptin tolerance in obese patients with diabetes. PMID- 21617187 TI - IL-13Ralpha1 expression on beta-cell-specific T cells in NOD mice. AB - OBJECTIVE: Immunotherapy using peptides from the beta-cell antigen GAD65 can preserve glucose homeostasis in diabetes-prone NOD mice; however, the precise mechanisms that arrest islet-reactive T cells remain unresolved. Our previous work revealed that a dominant GAD65 epitope contained two overlapping I-A(g7) restricted determinants, 524-538 and 530-543, with the former associated with amelioration of hyperglycemia. Here, we sought to discover whether p524-538 specific T cells could directly regulate islet-reactive T cells. RESEARCH DESIGN AND METHODS: Prediabetic NOD mice were used to determine the relationship between peptide p524-538-induced interleukin (IL)-13 and regulation of islet autoimmunity. Pancreatic lymph node (PLN) cells from mice at distinct stages of islet inflammation, peri-insulitis versus invasive insulitis, were harvested to establish the expression pattern of IL-13 receptor alpha1 (IL-13Ralpha1) on islet associated T cells. RESULTS: Peptide p524-538 preferentially induced IL-13 producing T cells that antagonized the release of gamma-interferon by spontaneously arising GAD65 autoimmunity, and recombinant human IL-13 inhibited proliferation of islet-reactive clonotypic T cells. A subset of CD4(+) T cells in NOD and NOD.BDC2.5 T cell receptor transgenic mice expressed functional IL 13Ralpha1, which induced phosphorylation of signal transducer and activator of transcription 6 in the presence of cognate cytokine. Notably, the number of IL 13Ralpha1(+) T cells was heightened in the PLN of young NOD mice when compared with older female counterparts with advanced insulitis. Immunization with p524 538 preserved IL-13Ralpha1 expression on PLN T cells. CONCLUSIONS: IL-13 may be important for regulating autoimmunity in the early stages of insulitis, and the loss of IL-13Ralpha1 on islet-reactive T cells may be a biomarker for fading regional immune regulation and progression to overt diabetes. PMID- 21617190 TI - Vitamin D for CRF or for dialysis patients: do we D-iscern? PMID- 21617188 TI - Mutations of the same conserved glutamate residue in NBD2 of the sulfonylurea receptor 1 subunit of the KATP channel can result in either hyperinsulinism or neonatal diabetes. AB - OBJECTIVE: Two novel mutations (E1506D, E1506G) in the nucleotide-binding domain 2 (NBD2) of the ATP-sensitive K(+) channel (K(ATP) channel) sulfonylurea receptor 1 (SUR1) subunit were detected heterozygously in patients with neonatal diabetes. A mutation at the same residue (E1506K) was previously shown to cause congenital hyperinsulinemia. We sought to understand why mutations at the same residue can cause either neonatal diabetes or hyperinsulinemia. RESEARCH DESIGN AND METHODS: Neonatal diabetic patients were sequenced for mutations in ABCC8 (SUR1) and KCNJ11 (Kir6.2). Wild-type and mutant K(ATP) channels were expressed in Xenopus laevis oocytes and studied with electrophysiological methods. RESULTS: Oocytes expressing neonatal diabetes mutant channels had larger resting whole-cell K(ATP) currents than wild-type, consistent with the patients' diabetes. Conversely, no E1506K currents were recorded at rest or after metabolic inhibition, as expected for a mutation causing hyperinsulinemia. K(ATP) channels are activated by Mg nucleotides (via SUR1) and blocked by ATP (via Kir6.2). All mutations decreased channel activation by MgADP but had little effect on MgATP activation, as assessed using an ATP-insensitive Kir6.2 subunit. Importantly, using wild-type Kir6.2, a 30-s preconditioning exposure to physiological MgATP concentrations (>300 umol/L) caused a marked reduction in the ATP sensitivity of neonatal diabetic channels, a small decrease in that of wild-type channels, and no change for E1506K channels. This difference in MgATP inhibition may explain the difference in resting whole-cell currents found for the neonatal diabetes and hyperinsulinemia mutations. CONCLUSIONS: Mutations in the same residue can cause either hyperinsulinemia or neonatal diabetes. Differentially altered nucleotide regulation by NBD2 of SUR1 can explain the respective clinical phenotypes. PMID- 21617191 TI - Sirolimus-based regimen is associated with decreased expression of glomerular vascular endothelial growth factor. AB - BACKGROUND: Sirolimus (SRL) is a potent immunosuppressant used in organ transplantation. It is known to decrease vascular endothelial growth factor (VEGF) synthesis, making it an interesting treatment option for transplant patients who develop Kaposi sarcoma or other malignant diseases. Because VEGF plays a key role in glomerular function and vascular remodelling, we determined the effect of SRL on renal VEGF expression. METHODS: Using immunohistochemistry and quantitative image analysis, we examined renal VEGF expression in routine kidney biopsies performed at 1 year post-transplant in the CONCEPT study, a prospective randomized study comparing a cyclosporine (CsA)-based regimen to a SRL-based regimen in association with mycophenolate mofetil (MMF). RESULTS: A total of 74 patients were included in this substudy; 35 were randomized to the CsA group and 39 to the SRL group. Using continuous variables, the mean percentage of glomerular VEGF expression at Week 52 was significantly lower in the SRL group (14.7 +/- 13%) compared to CsA group (21.2 +/- 14%: P = 0.02). The percentage of glomerular VEGF expression at Week 52 was not influenced by recipient or donor age, gender, renal function, CsA dose, CsA blood level, SRL dose or SRL blood level. It was significantly lower in patients with a proteinuria over versus below 0.5 g/day (11.58 +/- 7.9 versus 19.45 +/- 15.53; P = 0.036). CONCLUSIONS: There is emerging evidence that the VEGF system can play either a beneficial or a detrimental role depending on the specific pathologic situations. Therefore, modulating the renal VEGF axis by using an SRL-based regimen may influence the evolution of kidney injury associated with renal transplantation. PMID- 21617192 TI - Toll-like receptor 3 ligands induce CD80 expression in human podocytes via an NF kappaB-dependent pathway. AB - BACKGROUND: Recent studies suggest that CD80 (also known as B7.1) is expressed on podocytes in minimal-change disease (MCD) and may have a role in mediating proteinuria. CD80 expression is known to be induced by Toll-like receptor (TLR) ligands in dendritic cells. We therefore evaluated the ability of TLR to induce CD80 in human cultured podocytes. METHODS: Conditionally immortalized human podocytes were evaluated for TLR expression. Based on high expression of TLR3, we evaluated the effect of polyinosinic-polycytidylic acid (polyIC), a TLR3 ligand, to induce CD80 expression in vitro. RESULTS: TLR1-6 and 9 messenger RNA (mRNA) were expressed in podocytes. Among TLR ligands 1-9, CD80 mRNA expression was significantly induced by polyIC and lipopolysaccharide (TLR4 ligand) with the greatest stimulation by polyIC (6.8 +/- 0.7 times at 6 h, P < 0.001 versus control). PolyIC induced increased expression of Cathepsin L, decreased synaptopodin expression and resulted in actin reorganization which suggested a similar injury pattern as observed with lipopolyssaccharide. PolyIC induced type I and type II interferon signaling, nuclear factor kappa B (NF-kappaB) activation and the induction of CD80 expression. Knockdown of CD80 protected against actin reorganization and reduced synaptopodin expression in response to polyIC. Dexamethasone, a corticosteroid commonly used to treat MCD, also blocked both basal and polyIC-stimulated CD80 expression, as did inhibition of NF-kappaB. CONCLUSIONS: Activation of TLR3 on cultured human podocytes induces CD80 expression and phenotypic change via an NF-kappaB-dependent mechanism and is partially blocked by dexamethasone. These studies provide a mechanism by which viral infections may cause proteinuria. PMID- 21617193 TI - Polymorphism of the renalase gene in end-stage renal disease patients affected by hypertension. AB - BACKGROUND: Renalase is a novel flavin adenine dinucleotide-dependent amine oxidase that is secreted by the kidney. It circulates in the blood and modulates the cardiac function and systemic blood pressure. Insufficiency of renalase in patients with chronic kidney disease may explain the frequent occurrence of hypertension among patients with end-stage renal disease (ESRD) and an increased risk of cardiovascular events in this group. The aim of the study was to assess the relationship of two renalase gene polymorphisms with hypertension in dialysed patients. METHODS: Rs2576178 polymorphism was genotyped in 369 patients, rs10887800 polymorphism was genotyped in 421 dialysed patients, using polymerase chain reaction (PCR) and subsequent cleavage with Msp I and Pst I restriction endonucleases. RESULTS: Genotype distribution and allele frequencies of rs2576178 polymorphism were compared in the following subgroups of patients: dialysed patients with hypertension: ESRD HY + (n = 200) and dialysed patients without hypertension: ESRD HY - (n = 169). There was a significant difference in the frequency of the G allele carriers. G allele carriers were associated with a 1.55 times higher risk of hypertension [odds ratio (OR) = 1.55; 95% confidence interval (CI): 1.023-2.357, P = 0.039]. Distribution of genotypes and frequencies of alleles of rs10887800 polymorphism were compared in the following subgroups of patients: ESRD HY + (n = 278) and ESRD HY - (n = 143). The G allele carriers were recognized with a significantly higher frequency in ESRD HY + patients (0.46 in ESRD HY + versus 0.37 in ESRD HY - ) [OR = 1.76; 95% CI: (1.159-2.667, P = 0.008)]. CONCLUSIONS: Our results are the first to suggest an association between renalase gene polymorphisms analysed and hypertension in dialysed patients. It may be an important step towards gaining a deeper insight into cardiovascular pathophysiology. Furthermore, it might provide an optimal treatment and better prognosis for patients with chronic kidney disease. PMID- 21617194 TI - Acute pulmonary oedema in chronic dialysis patients admitted into an intensive care unit. AB - BACKGROUND: Acute pulmonary oedema (APO) in patients undergoing chronic dialysis (CD), a common cause of hospital admission in this population, is poorly documented. The objective of this study was to determine the causes, profile, clinical course and outcomes of APO in CD patients admitted in an intensive care unit (ICU). METHODS: Medical charts of all CD patients consecutively admitted for APO in the renal ICU of the Tenon Hospital (Paris, France) between January 2000 and December 2007 were considered. Data collection included patient characteristics, etiologic factors for chronic renal failure and co-morbidities, past history of APO, precipitating factors, clinical evolution and outcomes. RESULTS: Of the 112 files considered, 102 (65% men) were included in the final analysis. Patients were aged 20-88 years and had been dialysed for a median duration of 2 years. Hypertension (36.3%), chronic glomerulonephritis (25.5%) and diabetes mellitus (17.6%) were the main etiologic factors of chronic renal failure; 38.2% had a past history of APO. Acute pulmonary infection (26%), excessive interdialytic weight gain (25%) and inappropriate dry weight prescription (23%) were the leading causes of APO. The duration of hospitalization was <4 days in 60% of participants. Nine deaths (four being of cardiac origin) were recorded. Being referred from another hospital service was the main predictor of death. CONCLUSIONS: APO fuelled in part by chest infection, excessive interdialytic weight gain and inappropriate dry weight are important causes of hospitalization in CD patients. Mortality is high among those referred from other services usually in critical conditions. PMID- 21617195 TI - Intermediate bioelectrolyte changes after phospho-soda or polyethylene glycol precolonoscopic laxatives in a population undergoing health examinations. AB - BACKGROUND: Colonoscopy is a common procedure for diagnosing and screening colon cancer and other bowel-related diseases. Many studies have pointed out that using phospho-soda as a bowel preparation can cause obvious electrolyte abnormalities or acute kidney injury. Nonetheless, there are few studies related to its prevalence and risk factors in the population undergoing health examinations. Our aim was to compare the biochemical and electrolyte changes after using two commonly used bowel preparation regimens in this population. METHODS: In this retrospective study, we collected data about participants who, before a screening colonoscopy, used oral phospho-soda laxatives in 2006, and those who used polyethylene glycol-based laxatives in 2005. Several serum biochemical and electrolyte profiles were compared between the two groups. Additional risk factors of hyperphosphatemia, a well-known side effect of phospho-soda, were also derived. RESULTS: We enrolled a total of 2270 participants (1321 in 2005; 1449 in 2006). The basic demographic data of the two groups were not statistically different. Nonetheless, between the two groups, some serum biochemical and electrolytic data differed significantly: in those using oral phospho-soda laxatives, we found a higher prevalence of hyperuricemia, hypocalcemia, hypokalemia, hypernatremia and hyperphosphatemia. Further analyses showed that using oral phospho-soda laxatives was a risk factor for hyperphosphatemia; conversely, being male was a protective factor. CONCLUSION: Oral phospho-soda laxatives indeed influence the biochemical and electrolyte profiles of persons undergoing health examinations. One should be careful when interpreting bioelectrolytic data while using phospho-soda as a bowel preparation. PMID- 21617196 TI - Lupus nephritis: prolonged immunoadsorption (IAS) reduces proteinuria and stabilizes global disease activity. AB - BACKGROUND: Systemic lupus erythematosus (SLE) is characterized by pathogenic autoantibodies, which can be removed by extracorporeal procedures. While previous studies have shown short-term efficacy of immunoadsorption (IAS) in SLE, no information on long-term benefit and safety is available. METHODS: IAS was offered to patients with highly active renal disease when conventional therapy had failed. Eleven patients entered the prolonged IAS programme and were followed for up to 10 years (mean 6.4 +/- 3.5). Efficacy of IAS was determined by reduction in proteinuria (primary outcome), global disease activity [SLE Disease Activity Index (SLEDAI)] and anti-double-stranded DNA (anti-dsDNA) levels (secondary outcomes). Full/partial remission was defined as <= 0.5/<= 1.0 g/day for proteinuria, <= 5/<= 8 for SLEDAI and <= 25/<= 50 IU/mL for anti-dsDNA levels. We further assessed flares, infections, malignancies and procedure related adverse events. RESULTS: Short-term IAS (<= 1 year) resulted in a significant reduction of proteinuria (9.2 +/- 3.7 to 2.3 +/- 2.4, P = 0.0001), disease activity (SLEDAI 19 +/- 8 to 4 +/- 2, P = 0.0004) and dsDNA levels (168 +/- 205 to 45 +/- 34, P = 0.001). In patients without remission after 1 year (n = 5), prolonged IAS decreased proteinuria from 4.3 +/- 2.4 to 0.5 +/- 0.4 g/day, P = 0.02. At the end of observation, complete remission in proteinuria was achieved in seven patients (64%) and partial remission in two (18%) additional patients. One patient flared and was discontinued; in all other patients, disease activity and anti-dsDNA stabilized at remission levels. Flares (0.28 +/- 0.30) and infections (0.66 +/- 0.70 per patient/year) were relatively uncommon; no malignancies, anaphylactic or orthostatic adverse events were observed. CONCLUSION: IAS is effective in short-term use but prolonged IAS can provide additional therapeutic benefit while showing an acceptable safety profile. The vast majority of initially therapy-refractory patients met the remission criteria at the end of observation. PMID- 21617197 TI - Effect of different immunosuppressive regimens on the evolution of distinct metabolic parameters: evidence from the Symphony study. AB - BACKGROUND: The metabolic syndrome (MS) is an important risk factor for graft dysfunction and patient death after renal transplantation. The aim of this sub analysis of the Symphony study was to assess the progression of the laboratory parameters associated with MS in the first year after transplantation. METHODS: Data collected from the Symphony study were used; 1645 patients were randomized to receive standard-dose cyclosporine (Stand-CsA), low-dose cyclosporine (Low CsA), tacrolimus (Low-Tac) or sirolimus (Low-SRL), in addition to mycophenolate mofetil (MMF) and corticosteroids. Data were collected for levels and progression over the first year post-transplantation of systolic and diastolic blood pressure, uric acid, triglycerides, low-density lipoprotein (LDL) cholesterol, high-density lipoprotein (HDL) cholesterol and fasting glucose levels by treatment arm. RESULTS: The low-SRL group had significantly higher levels of triglycerides and LDL. The two CsA arms were associated with the highest uric acid levels at each time point. There were no significant differences in overall levels or changes in glucose or HDL. Patients in the standard-CsA arm had significantly higher diastolic blood pressure than those in the Low-SRL and Low Tac arms. Systolic blood pressure was higher in the Low-CsA arm than in the Low Tac arm. The use of antihypertensive and antidiabetic agents was similar between the treatment arms. In the Low-SRL arm, more patients were treated with lipid lowering therapy. Mean daily steroid doses were the highest in the Low-SRL arm. CONCLUSIONS: This sub-analysis demonstrates that there is a difference in metabolic parameters between immunosuppressive groups. CsA therapy was associated with the highest values of uric acid and systolic and diastolic blood pressure. Patients on SRL therapy had the worst lipaemic control. A possible effect of Tac on new-onset diabetes could not be excluded. PMID- 21617198 TI - Testosterone deficiency is a cause of anaemia and reduced responsiveness to erythropoiesis-stimulating agents in men with chronic kidney disease. AB - BACKGROUND: Hypogonadism or testosterone deficiency is a prevalent condition in men with chronic kidney disease (CKD). Testosterone stimulates erythropoiesis via production of haematopoietic growth factors and possible improvement of iron bioavailability. We hypothesized that testosterone deficiency predisposes to anaemia and reduced responsiveness to erythropoiesis-stimulating agents (ESAs) in CKD men. Materials and methods. We studied associations between endogenous testosterone and haemoglobin in 239 ESA-naive nondialysed CKD Stages 1-5 male patients. Additionally, we studied associations between endogenous testosterone levels and ESA dose (U/kg/week) in 126 ESA-treated men undergoing haemodialysis (HD). RESULTS: Among ESA-naive males, patients with anaemia presented lower testosterone values. Endogenous testosterone was negatively associated with haemoglobin levels in uni- and multivariate models. Testosterone-deficient patients (total testosterone <10 nmol/L) were 5.3 (95% confidence interval 2.2 12.5) times more likely to be anaemic (Hb < 13.0 g/dL) than testosterone sufficient patients. In ESA-treated men undergoing HD, higher ESA doses (above the median value of 121 IU/kg body weight/week) are associated with lower testosterone levels and higher percentage of hypochromic red blood cells (RBC). The inverse association between testosterone levels and ESA doses persisted after multivariate adjustment for age, sex hormone-binding globulin, comorbidities, C reactive protein and s-albumin but was lost after further adjustment for iron medication and hypochromic RBC. CONCLUSIONS: Hypogonadism may be an additional cause of anaemia and reduced ESA responsiveness in men with CKD. Our results raise the possibility that restoration of testosterone levels in hypogonadal CKD males may translate into lower prevalence of anaemia and better ESA responsiveness. PMID- 21617199 TI - Renal transplant recipients have elevated frequencies of circulating myeloid derived suppressor cells. AB - BACKGROUND: Cancer, particularly cutaneous squamous cell carcinoma (SCC), is a major cause of mortality in renal transplant recipients (RTRs). Myeloid-derived suppressor cells (MDSC) play a central role in suppressing cancer immunosurveillance but their potential mobilisation in RTRs and levels relative to those of other immunoregulatory dendritic cell (DC) populations have not been analysed. METHODS: The circulating frequencies of MDSC and DC were analysed by multicolour flow cytometry in immunocompetent patients without (n = 13) or with (ICI-SCC(Pos), n = 14) current SCC, normal donors (NDs, n = 34), chronic kidney disease patients (CKD patients, n = 22) and RTRs (n = 31). RESULTS: Compared to NDs, RTRs had significantly elevated levels of both CD14(Neg) and CD14(Pos) MDSC subsets (P < 0.001), while CKD patients and ICI-SCC(Pos) had significantly elevated levels of only the CD14(Neg)-MDSC subset. DC frequencies were significantly decreased in RTRs and CKD patients but were at normal levels in ICI SCC(Pos). The MDSC/DC ratio was significantly elevated (P < 0.05) in RTRs (median = 5.7), CKD patients (median = 3.2) and ICI-SCC(Pos) (median = 3.5) relative to NDs (median = 0.7). The use of immunosuppressive drugs in CKD patients and past/current occurrence of SCC in RTRs was associated with significantly increased CD14(Neg)-MDSC frequencies. MDSC enriched from RTRs, when co-cultured with activated NDs T cells significantly suppressed extracellular IL-10 levels and can, when activated with formyl-methionyl-leucyl-phenylalanine, inhibit T cell proliferation. CONCLUSIONS: RTRs, CKD patients and ICI-SCC(Pos) have increased MDSC frequencies and MDSC/DC ratios. These changes may impact on cancer immunosurveillance. Therefore, MDSC represent both a potential therapeutic target and prognostic marker in these patients, with respect to the development of SCC and other malignancies. PMID- 21617200 TI - Toll-like receptor 4 potentiates Ca2+-dependent secretion of electrolytes from swine tracheal glands. AB - Airway surface fluids are mainly secreted from submucosal glands, and play important roles in the defense of airways via the up-regulation of mucociliary transport, resulting in an exclusion of many microbes or foreign substances. Although there are many articles concerning the importance of Toll-like receptors (TLRs) in airway immune systems, whether TLRs directly cooperate with tracheal submucosal glands to increase secretion remains unknown. We investigated the effects of ligands of the three TLR subtypes (TLR2, TLR3, and TLR4) on the physiologic secretion of electrolytes by using a patch-clamp technique. Among these TLRs, only the TLR4 ligand, LPS, showed potentiating effects on acetylcholine (ACh)-induced ionic currents in a dose-dependent manner. These potentiating effects were completely abolished by pretreatment with a specific TLR4 antagonist or the anti-TLR4 antibody. LPS per se exerted no appreciable effect on baseline currents. Next, we demonstrated the abundant expression of TLR4 in submucosal gland acinar cells by using immunofluorescent staining and RT PCR. Furthermore, we revealed that both nitric oxide synthase inhibitors and cyclic guanosine monophosphate (cGMP)-dependent protein kinase (cGK) inhibitors abolished the LPS-induced potentiating effects completely. Analyses of fluorescence intensities, using an intracellular nitric oxide (NO) indicator, demonstrated that LPS could further increase the ACh-induced synthesis of NO. These findings suggest that TLR4 takes part in airway mucosal defense systems as a unique exogenous potentiator of electrolyte-water secretion from submucosal gland acinar cells, and that NO/cGMP/cGK signaling is involved in this rapid TLR4 signaling pathway. PMID- 21617201 TI - Polycyclic aromatic hydrocarbons impair function of beta2-adrenergic receptors in airway epithelial and smooth muscle cells. AB - Incomplete combustion produces a pollutant mixture that includes polycyclic aromatic hydrocarbons (PAHs). Previous work by the Columbia Center for Children's Environmental Health (CCCEH) and others linked exposure to PAH with symptoms of asthma and other adverse health effects in young children. Inhaled beta(2) adrenergic agonists are mainstays in the treatment of reactive airway diseases. These exogenous catecholamines engage membrane-bound beta(2)-adrenergic receptors (beta(2)AR) on airway epithelial and smooth muscle cells to cause airway dilation. We hypothesized that exposure to PAH might similarly interfere with the function of beta(2)AR in airway epithelial or smooth muscle cells, reducing the efficacy of a medication important for the treatment of asthma symptoms. A PAH mixture was devised, based on ambient levels measured prenatally among a cohort of pregnant women participating at the CCCEH. Primary airway epithelial and smooth muscle cells were exposed to varying concentrations of the PAH mixture, and expression, function, and signaling of beta(2)AR were assessed. Murine tracheal epithelial cells and human airway smooth muscle cells, after exposure to a PAH mixture, exhibited reduced expression and function of beta(2)AR. These findings support our hypothesis that environmentally relevant PAHs can impede beta(2)AR-mediated airway relaxation, and suggest a new paradigm where air pollutants not only contribute to the pathogenesis of childhood asthma, but also diminish responsiveness to standard therapy. PMID- 21617202 TI - Urokinase plasminogen activator regulates pulmonary arterial contractility and vascular permeability in mice. AB - The concentration of urokinase plasminogen activator (uPA) is elevated in pathological settings such as acute lung injury, where pulmonary arterial contractility and permeability are disrupted. uPA limits the accretion of fibrin after injury. Here we investigated whether uPA also regulates pulmonary arterial contractility and permeability. Contractility was measured using isolated pulmonary arterial rings. Pulmonary blood flow was measured in vivo by Doppler and pulmonary vascular permeability, according to the extravasation of Evans blue. Our data show that uPA regulates the in vitro pulmonary arterial contractility induced by phenylephrine in a dose-dependent manner through two receptor-dependent pathways, and regulates vascular contractility and permeability in vivo. Physiological concentrations of uPA (<=1 nM) stimulate the contractility of pulmonary arterial rings induced by phenylephrine through the low-density lipoprotein receptor-related protein receptor. The procontractile effect of uPA is independent of its catalytic activity. At pathophysiological concentrations, uPA (20 nM) inhibits contractility and increases vascular permeability. The inhibition of vascular contractility and increase of vascular permeability is mediated through a two-step process that involves docking to N methyl-d-aspartate receptor-1 (NMDA-R1) on pulmonary vascular smooth muscle cells, and requires catalytic activity. Peptides that specifically inhibit the docking of uPA to NMDA-R, or the uPA variant with a mutated receptor docking site, abolished both the effects of uPA on vascular contractility and permeability, without affecting its catalytic activity. These data show that uPA, at concentrations found under pathological conditions, reduces pulmonary arterial contractility and increases permeability though the activation of NMDA-R1. The selective inhibition of NMDAR-1 activation by uPA can be accomplished without a loss of fibrinolytic activity. PMID- 21617203 TI - A sphingosine 1-phosphate 1 receptor agonist modulates brain death-induced neurogenic pulmonary injury. AB - Lung transplantation remains the only viable therapy for patients with end-stage lung disease. However, the full utilization of this strategy is severely compromised by a lack of donor lung availability. The vast majority of donor lungs available for transplantation are from individuals after brain death (BD). Unfortunately, the early autonomic storm that accompanies BD often results in neurogenic pulmonary edema (NPE), producing varying degrees of lung injury or leading to primary graft dysfunction after transplantation. We demonstrated that sphingosine 1-phosphate (S1P)/analogues, which are major barrier-enhancing agents, reduce vascular permeability via the S1P1 receptor, S1PR1. Because primary lung graft dysfunction is induced by lung vascular endothelial cell barrier dysfunction, we hypothesized that the S1PR1 agonist, SEW-2871, may attenuate NPE when administered to the donor shortly after BD. Significant lung injury was observed after BD, with increases of approximately 60% in bronchoalveolar lavage (BAL) total protein, cell counts, and lung tissue wet/dry (W/D) weight ratios. In contrast, rats receiving SEW-2871 (0.1 mg/kg) 15 minutes after BD and assessed after 4 hours exhibited significant lung protection (~ 50% reduction, P = 0.01), as reflected by reduced BAL protein/albumin, cytokines, cellularity, and lung tissue wet/dry weight ratio. Microarray analysis at 4 hours revealed a global impact of both BD and SEW on lung gene expression, with a differential gene expression of enriched immune-response/inflammation pathways across all groups. Overall, SEW served to attenuate the BD-mediated up-regulation of gene expression. Two potential biomarkers, TNF and chemokine CC motif receptor like 2, exhibited gene array dysregulation. We conclude that SEW-2871 significantly attenuates BD-induced lung injury, and may serve as a potential candidate to improve human donor availability. PMID- 21617204 TI - An epidemic of oropharyngeal squamous cell carcinoma (OSCC) due to human papillomavirus (HPV) infection and aspects of treatment and prevention. AB - In Europe and the US the incidence of oropharyngeal squamous cell carcinoma (OSCC), where tonsillar and tongue base cancer dominate, has increased and we propose this is due to an epidemic of human papillomavirus (HPV) infection. We base our hypothesis on two main findings. Firstly, in the past decade numerous reports have shown the presence of HPV in OSCC, and that HPV-positive OSCC has a significantly better response to therapy than HPV-negative OSCC. As a consequence of these findings, the International Agency for Research against Cancer (IARC) has acknowledged HPV, in addition to smoking and alcohol, as a risk factor for OSCC. Secondly, we and others have demonstrated a considerable and significant increase in the proportion of HPV-positive tonsillar and tongue base cancer/OSCC in recent decades. In conclusion, we suggest that the increased incidence of OSCC is due to an HPV epidemic. In addition, since patients with HPV-positive OSCC have a better response to therapy than those with HPV-negative OSCC, we put forward that it is important to individualize therapy on the basis of HPV status and other markers in order to obtain optimal survival with the least side effects. Finally, we suggest that the possibility to vaccinate against HPV positive OSCC should be examined. PMID- 21617205 TI - Synthesis and evaluation of a novel lipophilic folate receptor targeting ligand. AB - BACKGROUND: Folate receptor (FR)-targeted liposomes have been investigated as delivery vehicles for anticancer drugs. A novel lipophilic FR ligand, folate glutathione-polyethyleneglycol-distearoyl phosphatidylethanolamine (F-GSH-PEG DSPE), was synthesized, incorporated into liposomes and evaluated for FR targeting efficiency. These liposomes were then evaluated as carriers of the chemotherapy agent vincristine (VIN). MATERIALS AND METHODS: F-GSH-PEG-DSPE was synthesized and FR-targeted liposomes loaded with either calcein (F-L-Calcein) or VIN (F-L-VIN) were prepared by thin film hydration followed by polycarbonate membrane extrusion and, in the case of VIN, by remote loading. To assess liposome stability, the uptake of F-L-VIN in KB (FR+) cancer cells was measured after storage under 4 degrees C for 3 months. Comparative pharmacokinetic studies were carried out with F-L-VIN and L-VIN (non-targeted control liposomes). RESULTS: F-L Calcein showed significantly higher cellular uptake in KB cells compared to non targeted liposomes. In addition, F-L-VIN showed enhanced cytotoxicity in KB cells in vitro compared to control liposomes. Pharmacokinetic parameters indicated that both F-L-VIN and control liposomes had higher area under the curve (AUC), mean residence time (MRT), elimination half life (t1/2-beta) and lower total body clearance (CL) than those of free VIN, while there were no significant differences between these liposomal formulations. CONCLUSION: F-GSH-PEG-DSPE is effective as a novel ligand for the synthesis of FR-targeted liposomes. PMID- 21617206 TI - Direct effects of pure nicotine, cigarette smoke extract, Swedish-type smokeless tobacco (Snus) extract and ethanol on human normal endothelial cells and fibroblasts. AB - The adverse health effects of cigarette smoking are well established including the increased risk of various types of cancer. In this study, the direct effects of ethanol, pure nicotine, cigarette smoke extract and Swedish type smokeless tobacco (Snus) extract on normal cells were investigated. MATERIALS AND METHODS: Primary normal adult human endothelial cells and fibroblasts at early passage were used. Upon exposure to pure nicotine, cigarette smoke extract, Snus extract and ethanol, these cells were assessed for DNA synthesis, gene expression profile and cellular morphology. RESULTS: Normal human fibroblasts and endothelial cells have unique gene expression profiles. The effects of treatment with ethanol and nicotine from different sources was more prominent in endothelial cells than fibroblasts. The combination of alterated gene expressions and strongly inhibited DNA synthesis was only detected in cells exposed to smoke extract. In the presence and absence of ethanol, pure nicotine and Snus extract induced abnormalities in the cytoplasm without any significant degree of cell death. With similar doses of nicotine and ethanol, the additional components in smoke extract had a dominant effect. The smoke extract induced vast cellular abnormalities and massive cell death. CONCLUSION: Cigarette smoke induced massive cell death and various abnormalities at cellular and molecular levels in surviving endothelial cells and fibroblasts. The combination of genomic alterations and the chronic inflammatory microenvironment induced from massive cell death, will potentially promote tumourigenesis and various diseases in cigarette smokers. PMID- 21617207 TI - Circulating human prostate cancer cells from an orthotopic mouse model rapidly captured by immunomagnetic beads and imaged by GFP expression. AB - Circulating tumor cells (CTCs) are potential precursors of metastasis. They are also of use in diagnosing malignancy and for prognostic purposes. Our laboratory has previously isolated CTCs from orthotopic nude mouse models of human prostate cancer cells where the PC-3 cancer cells express green fluorescent protein (GFP). It was found that orthotopic tumors produced CTCs and not subcutaneous tumors, which may explain why orthotopic tumors metastasize and subcutaneous tumors do not. However, in this previous study, CTCs were observed only after culture. In the present study, using the GFP-expressing PC-3 orthotopic model and immunomagnetic beads coated with anti-epithelial cell adhesion molecule (EpCAM) and anti-prostate specific membrane antigen (PSMA), GFP-expressing CTC were isolated within 15 minutes and were readily visualized by GFP fluorescence. It was possible to immediately place the immunomagnetic-bead-captured GFP-expressing PC-3 CTCs in 3-dimensional sponge cell culture, where they proliferated. The combination of GFP expression and the use of immunomagnetic beads is a very powerful method to obtain CTCs for either immediate analysis or for biological characterization in vivo or in 3-dimensional culture. PMID- 21617208 TI - Quantitative PCR and immunohistochemical analyses of HMGB1 and RAGE expression in canine disseminated histiocytic sarcoma (malignant histiocytosis). AB - BACKGROUND: Disorders of histiocytic origin affecting humans and dogs share various similarities. Canine disseminated histiocytic sarcoma (DHS) (formerly known as malignant histiocytosis) is an aggressive neoplasm of interstitial dendritic cells (DCs). The receptor for glycation end products (RAGE) and the high mobility group box1 protein (HMGB1) have been shown to be required for the maturation and migration of DCs. Thus, deregulation of the expression of these genes could have a major effect on the progression of histiocytic disorders. MATERIALS AND METHODS: Neoplastic canine DHS samples and non-neoplastic control samples were analysed immunohistochemically and via real-time PCR. RESULTS: Significant down-regulation of RAGE in the lung tumour samples and down regulation of HMGB1 in the lung, lymph node and spleen tumour samples were detected compared to their non-neoplastic counterparts. CONCLUSION: RAGE and HMGB1 expression down-regulation in canine DHS points to a role in the progression of histiocytic disorders. PMID- 21617209 TI - Anticancer effect of dihydroartemisinin (DHA) in a pancreatic tumor model evaluated by conventional methods and optical imaging. AB - BACKGROUND: Dihydroartemisinin (DHA) inhibits the growth of certain cancer cells and xenograft tumors. Further understanding of the molecular mechanisms and genetic participants that govern the antineoplastic effects of DHA is necessary. The anticancer effects of DHA and its underlying mechanisms in pancreatic cancer and the efficacy in animal models by noninvasive optical imaging were evaluated. MATERIALS AND METHODS: Combined with cell/tumor growth assays, flow cytometric analysis, and Hoechst staining, the effect of DHA was investigated using the pancreatic cancer cell line BxPc3-RFP stably expressing red fluorescence protein and in vitro/in vivo optical imaging. Proteins that regulate proliferation (PCNA), apoptosis (Bax and Bcl-2), and angiogenesis (vascular endothelial growth factor (VEGF)) were evaluated in cell and tumor samples by Western blotting and immunohistochemical analyses. RESULTS: DHA inhibited the proliferation and viability of cells in a dose-dependent manner and induced apoptosis. We observed down-regulation of PCNA and Bcl-2, and up-regulation of Bax. VEGF was down regulated by DHA in cells under normoxic, but not hypoxic, conditions. Fluorescence intensity emitted from cells and tumors correlated linearly with cell count and tumor burden, respectively. CONCLUSION: DHA inhibits cell and tumor growth by interfering with cell proliferation and inducing apoptosis. The antiangiogenic effect of DHA appears to be a complicated process. Optical imaging supports the real-time assessment of DHA efficacy in a preclinical model and comprehensive analysis substantiates that DHA is a potential candidate for pancreatic cancer therapy. PMID- 21617210 TI - Pretreatment HIF-1alpha and GLUT-1 expressions do not correlate with outcome after preoperative chemoradiotherapy in rectal cancer. AB - The aim of the present study was to investigate hypoxia-inducible factor 1alpha (HIF-1alpha) and glucose transporter-1 (GLUT-1) expressions as predictors of response and survival after chemoradiotherapy in pretreatment biopsy specimens from patients with rectal cancer. MATERIALS AND METHODS: The immunohistological expressions of HIF-1alpha and GLUT-1 were investigated in pretreatment biopsies from 86 patients with rectal cancer receiving long course preoperative chemoradiotherapy. The immunohistological stainings were scored semi quantitatively (percentage of stained cells and staining intensity), and an immunoreactive score was calculated. The response to the chemoradiotherapy was assessed by the Mandard Tumour Regression Grade system (TRG). RESULTS: No association was found between HIF-1alpha or GLUT-1 and clinicopathological variables. HIF-1alpha and GLUT-1 expression had no predictive impact regarding response to chemoradiotherapy measured by TRG and was not associated with overall survival. CONCLUSION: The present study did not suggest any predictive or prognostic value of pretreatment HIF-1alpha or GLUT-1 expression in patients with rectal cancer treated with preoperative chemoradiotherapy. PMID- 21617211 TI - Heterogeneity of metalloproteinase expression in colorectal cancer - relation of molecular findings to basic morphology. AB - AIM: To clarify the correlation between morphological features and the mRNA expression of a disintegrin and metalloproteinases (ADAMs) and matrix metalloproteinases (MMPs) in colorectal cancer. MATERIALS AND METHODS: In 29 colorectal tumors, cancer cells were isolated by laser microdissection, and the mRNA expression of metalloproteinases was compared between the tumor center and the invasive front. RESULTS: Regarding the central region as the standard, the mRNA levels of ADAM-12, MMP-1, -2, -7, and -9 at the invasive front were up regulated. The degree of up-regulation was significantly higher in solid cancer nest (SCN)-positive tumors than in SCN-negative tumors for the mRNA levels of ADAM-12, MMP-2, -7, and - 9 (SCN-positive, median 5.1-, 3.9-, 9.9-, and 2.7-fold; SCN-negative 2.0-, 1.0-, 2.4-, and 0.8-fold; p<0.05, respectively). CONCLUSION: Up-regulation of the mRNA expression of particular metalloproteinases was significantly associated with the SCN at the leading edge of colorectal tumors. PMID- 21617212 TI - Overexpression of the TXNDC5 protein in non-small cell lung carcinoma. AB - Thioredoxin domain containing protein 5 (TXNDC5) is a member of the thioredoxin (Trx) domain-containing family of proteins that have been implicated in cancer progression. The expression of TXNDC5 in non-small cell lung carcinoma (NSCLC) tumours compared to patient-matched normal lung tissue was determined and cell line models were used to determine if expression was regulated by hypoxia. PATIENTS AND METHODS: Samples of tumour and normal lung tissue were taken during surgery and immediately frozen. The expression of TXNDC5 was determined by Western blotting and immunohistochemistry. To analyse the effect of hypoxia on TXNDC5 expression NSCLC cell lines were used. RESULTS: Tumours from 18/29 (62%) individuals exhibited an increase in TXNDC5 expression compared to normal lung tissue (p<0.05). TXNDC5 expression was not elevated by hypoxia. CONCLUSION: TXNDC5 is up-regulated in the majority of resected human NSCLC. Cell line data indicates that the expression of TXNDC5 in tumour cells is not regulated by hypoxia. PMID- 21617213 TI - Effect of hyperthermia on liver cell lines: important findings for thermal therapy in hepatocellular carcinoma. AB - BACKGROUND: Hepatocellular carcinoma is the fifth most common malignant tumour, with a high mortality rate. This study aimed to investigate the effect of hyperthermia on HepG2 and LX-1 cell lines to gain more information on thermal treatment of liver tumours. MATERIALS AND METHODS: The cell lines HepG2, LX-1 and their co-cultures were heated from 55 degrees C to 85 degrees C for different time spans. After heat exposure, metabolic activity was measured immediately, and after 24 h and 48 h using the 3-(4,5-dimethylthiazol-2-yl)-5-(3 carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) (MTS) test to assess how many cells had survived heating. RESULTS: Our results show highly significant differences between the temperature tolerance of HepG2 and LX-1 cells. Alone, HepG2 cells are most sensitive to heat-induced cell death, their sensitivity decreased with rising percentages of LX-1 cells in the co-culture. CONCLUSION: Our results suggest that the outcome of thermal cancer therapy is dependent on the temperature and the grade of fibrosis in the treated livers. PMID- 21617214 TI - Lectin histochemistry of metastasizing and non-metastasizing breast and colon cancer cells. AB - BACKGROUND: Glycosylation of the tumour cell surface is of importance in metastasis formation as indicated by lectin-binding studies. In particular, binding of the lectin HPA is associated with metastasis formation, both in clinical studies and in xenograft models of breast and colon cancer. Here we examined if there is an association between the HPA-positive glycotopes of metastasizing cancer cells and selectin-binding properties. MATERIALS AND METHODS: Glycotope expression of human breast and colon cancer cells (MCF7, T47D, HBL100, HT29, SW480) grown in culture and xenografted into SCID mice were investigated by histochemical analysis. RESULTS: HPA binding was observed in metastasizing breast and colon cancers and not in non-metastasizing ones. In colon cancer, E-selectin binding and expression of the selectin ligands CD15s and CA19-9 was higher in metastatic HT29 than in non-metastatic SW480 cells, especially when cells were grown in vitro. In breast cancer, E-selectin binding, CD15s and CA19-9 expression were independent of the metastatic potential. P Selectin binding was slightly higher in metastasizing breast cancer cells (MCF7, T47D) than in non-metastasizing HBL100 cells. CONCLUSION: Binding to E-selectin and expression of E-selectin ligands of colon cancer cells grown in vitro is associated with metastasis formation in a xenograft model. However, analysis of selectin ligands is of limited predictive value for the metastatic potential of breast cancer cells in our xenograft model. PMID- 21617215 TI - Structural rearrangements of trisomies are a risk marker of clinical progression in hyperdiploid multiple myeloma. AB - BACKGROUND: Hyperdiploid multiple myeloma (HMM) is being characterized by the presence of several trisomies and a low incidence of immunoglobulin heavy chain rearrangements. It has not well defined what specific steps are associated with disease progression. We present two patients that showed some primary trisomies rearranged as a step of cytogenetic and clinical progression. This prompted us to review cytogenetic results from all patients referred to our hospital to assess the importance of this phenomenon in HMM. PATIENTS AND METHODS: We carried out conventional cytogenetics in all patients. In four cases we also performed spectral karyotype (SKY) and arm-specific chromosome painting (ASP). RESULTS: We demonstrate that in two patients some primary trisomies became along the disease course structurally altered and this coincided with clinical progression. We observed this phenomenon in more than 60% of HMM cases diagnosed at our laboratory. CONCLUSION: We propose structural rearrangements of trisomies as a biological marker of progression in HMM. PMID- 21617216 TI - Correlation and coexpression of HIFs and NOTCH markers in NSCLC. AB - BACKGROUND: NOTCH and hypoxia pathways are both known to be highly involved in cancer. Because of the close interplay between both of these pathways, we investigated correlation and co-expression of molecules in these pathways. MATERIALS AND METHODS: In 335 unselected stage I-IIIA NSCLC patients, protein expressions of hypoxia inducible factor 1alpha (HIF1alpha), hypoxia inducible factor 2alpha (HIF2alpha), glucose transporter 1 (GLUT1), lactate dehydrogenase 5 (LDH5), carbonic anhydrase IX (CAIX), delta like 4 (DLL4), JAGGED1, NOTCH1 and NOTCH4, evaluated by immunohistochemistry, were correlated and co-expressions tested in tumor and stromal cells. RESULTS: HIF2alpha and LDH5 correlated moderately with DLL4, JAGGED1 and NOTCH4 in both tumor and stromal compartments (Spearman's r=0.16-0.33). The coexpression of HIF1alpha and NOTCH1 in tumor was significantly indicative of poor prognosis in univariate analysis. Hypoxia and NOTCH ligands and receptors were moderately correlated. CONCLUSION: The lack of appealing coexpression findings for HIF1alpha and NOTCH1 may be due to the way HIF1alpha directly influences NOTCH signalling without depending on an elevated NOTCH expression. PMID- 21617217 TI - NF-kappaB activation by peroxynitrite through IkappaBalpha-dependent phosphorylation versus nitration in colon cancer cells. AB - BACKGROUND: Peroxynitrite has been proposed to activate Nuclear Factor kappaappa beta (NF-kappaB) in a non-canonical or aberrant pathway and to contribute to pathology of many diseases. METHODS: The activation of NF-kappaB by peroxynitrite was assessed by Western blot, immunoprecipitation, RT-PCR, and image stream analysis. RESULTS: Our work showed that in HT-29 cancer cells, peroxynitrite can cause nitration of Inhibitory protein kappa B alpha (IkappaBalpha) at the expense of its phosphorylation. This led to a decrease in the degradation and re synthesis of IkappaBalpha. Similar findings were observed for mRNA levels assessed by RT-PCR. Exposure of HT-29 cells to p38 inhibitor SB202190, prior to stimulation, resulted in a dramatic decrease of IkappaBalpha kinase and IkappaBalpha phosphorylation and caused an increase of peroxynitrite-mediated nitration of IkappaBalpha, indicating that peroxynitrite may activate NF-kappaB via dual mechanism of IkappaBalpha phosphorylation which is p38 dependent, as well as IkappaBalpha nitration. CONCLUSION: Our findings demonstrate a possible interaction of the p38 pathway with the NF-kappaB pathway under peroxynitrite stimulation. PMID- 21617218 TI - Efficient down-regulation of CDK4 by novel lipid nanoparticle-mediated siRNA delivery. AB - BACKGROUND: Inhibition of cyclin-dependent kinases 4 (CDK4) activity by Small interfering RNA (siRNA) has been demonstrated as one of the promising approaches to treat cancer. MATERIALS AND METHODS: CDK4 siRNA was packaged in a lipid nanoparticle (LNP)-based delivery system that consists of an ionizable cationic lipid, helper lipid and polyethylene glycol (PEG)-lipid. The physical properties, including the size and surface charge of LNP-siRNA, were characterized by dynamic light scattering and zeta potential measurements. The biological activities of LNP-siRNA, including the cellular uptake, CDK4 expression and G(1) cell cycle arrest, in both HeLa cervical cancer and MDA-MB-468 breast cancer cells were evaluated by flow cytometry, confocal microscopy, quantitative reverse transcription PCR (qRT-PCR) and Western blotting, respectively. RESULTS: The new LNP-mediated siRNA delivery demonstrated enhanced cellular uptake. Compared with free siRNA and lipofectamine-formulated siRNA, cells treated by LNP-CDK4 siRNA exhibited significant G(1) cell cycle arrest, which was consistent with efficient down-regulation of CDK4 at both mRNA and protein levels. CONCLUSION: Gene silencing of CDK4 by LNP-siRNA offers an alternative strategy for CDK4-based cancer therapy. The new LNP can be used for efficient delivery of siRNA in vitro. PMID- 21617219 TI - Targeting of breast metastases using a viral gene vector with tumour-selective transcription. AB - BACKGROUND: Adeno-associated virus (AAV) vectors have significant potential as gene delivery vectors for cancer gene therapy. However, broad AAV2 tissue tropism results in nonspecific gene expression. MATERIALS AND METHODS: We investigated use of the C-X-C chemokine receptor type 4 (CXCR4) promoter to restrict AAV expression to tumour cells, in subcutaneous MCF-7 xenograft mouse models of breast cancer and in patient samples, using bioluminescent imaging and flow cytometric analysis. RESULTS: Higher transgene expression levels were observed in subcutaneous MCF-7 tumours relative to normal tissue (muscle) using the CXCR4 promoter, unlike a ubiquitously expressing Cytomegalovirus promoter construct, with preferential AAVCXCR4 expression in epithelial tumour and CXCR4-positive cells. Transgene expression following intravenously administered AAVCXCR4 in a model of liver metastasis was detected specifically in livers of tumour bearing mice. Ex vivo analysis using patient samples also demonstrated higher AAVCXCR4 expression in tumour compared with normal liver tissue. CONCLUSION: This study demonstrates for the first time, the potential for systemic administration of AAV2 vector for tumour-selective gene therapy. PMID- 21617220 TI - Experimental study of the anticancer effect of gemcitabine combined with sirolimus on chemically induced urothelial lesions. AB - BACKGROUND: The purpose of this study was to determine the efficacy of a combination of gemcitabine and sirolimus in a mouse model of invasive bladder cancer. MATERIALS AND METHODS: Gemcitabine (50 mg/kg) and sirolimus (1.5 mg/kg) were administered to animals previously exposed to N-butyl-N 4(hydroxybutyl)nitrosamine in drinking water. Tumour development was determined by histopathological evaluation. RESULTS: Both drugs were well tolerated by animals. The incidence of lesions in mice treated with gemcitabine was lower in comparison to those not treated, however this result was not statistically significant. The incidence of invasive bladder cancer in animals treated with sirolimus was statistically lower (20%) than in animals not treated (54%) (p=0.008). The results indicate that this drug combination has no statistical significance on the development of pre-neoplastic urothelial lesions and had only a minor impact on invasive bladder cancer incidence in mice. CONCLUSION: The combination of gemcitabine and sirolimus had only a marginal impact on invasive bladder cancer in a mouse model. PMID- 21617221 TI - p16INK4a methylation in serum as a follow-up marker for recurrence of colorectal cancer. AB - BACKGROUND: p16(INK4a) methylation present in the tumors of colorectal cancer (CRC) patients can be detected in their serum using quantitative methylation specific PCR (Q-MSP). To investigate the possibility that this technique could be applied to the monitoring for cancer recurrence in CRC patients, p16(INK4a) methylation in the serum of CRC patients during their follow-up period was evaluated. MATERIALS AND METHODS: Using Q-MSP on serum samples from 21 CRC patients undergoing surgery for primary CRC, the p16INK4a methylation score (p16(INK4a) MS) was evaluated one day before surgery and during the follow-up period. RESULTS: In the serum samples collected before primary resection, p16(INK4a) methylation was detected in 8 out of the 13 patients with same methylation in the tumor. The p16(INK4a) MS decreased within 2 weeks after surgery. Only two patients, who had the potential for recurrence, exhibited p16(INK4a) methylation in their serum. One month after surgery, in the patients with recurrence of tumor, a dramatic increase in p16(INK4a) MS was observed, while in the disease-free patients no methylation was seen continuously. CONCLUSION: p16(INK4a) MS could sensitively reflect the recurrence status and may be useful for identifying the presence of recurrence during the follow-up of CRC patients. PMID- 21617222 TI - Barley low molecular weight beta-glucan potently induces maturation of mouse dendritic cells. AB - BACKGROUND: Accumulating evidence indicates that non-toxic immunostimulants with strong differentiation/maturation-inducing activity for dendritic cells (DCs) might be useful for preventing or even curing cancer. MATERIALS AND METHODS: Mouse bone marrow (BM) cells were cultured in the presence of various glucans and their differentiation/maturation-inducing activities were compared by measuring cytokines secreted in the culture medium. RESULTS: Barley-derived beta-glucan with an average molecular weight of 2 kDa (BBG-Low) remarkably stimulated the formation of mature DCs from immature mouse DCs. The amount of interleukin-6 produced by sequential treatment of BM cells with granulocyte-macrophage colony stimulating factor and 10 MUg/mL of BBG-Low was approximately 30 times higher than that obtained by a similar sequential treatment using barley beta-glucan of 40-70 kDa instead of BBG-Low. CONCLUSION: BBG-Low induces the formation of mature DCs from immature DCs and suggests that BBG-Low will be useful as a potent nontoxic immunostimulator. PMID- 21617223 TI - The effect of PTEN on proliferation and drug-, and radiosensitivity in malignant glioma cells. AB - BACKGROUND: Deletions or mutations of the phosphatase and tensin homolog (PTEN) are frequently observed in malignant glioma and are responsible for progression of the disease. Since the molecule is a promising target for gene therapy, the effects of PTEN on glioma proliferation in combination with the anti-neoplastic agent, temozolomide, and ionizing radiation were investigated. MATERIALS AND METHODS: An adenoviral vector encoding PTEN was used. After infection, changes in proliferation, the cell cycle, as well as drug- and radiosensitivity were investigated. RESULTS: Expression of PTEN led to a 1.21-fold prolongation of the doubling time of the cells. It reduced G(1) and increased G(2)/M populations. Forced PTEN expression conferred sensitivity to temozolomide and/or ionizing radiation. CONCLUSION: In addition to counteracting cell proliferation, expression of PTEN presented advantages in the chemo- and radiosensitivity of glioma cells. Methods for up-regulation of PTEN may have a role in increasing the efficacy of current adjuvant therapies. PMID- 21617224 TI - Zeranol induces cell proliferation and protein disulfide isomerase expression in mammary gland of ACI rat. AB - BACKGROUND: Zeranol is a non-steroidal anabolic growth promoter with potent estrogenic activity that is widely used as a growth promoter in the US beef industry. Consumption of beef derived from Zeranol-implanted cattle may be a risk factor for breast cancer. Protein disulfide isomerase (PDI) has been studied extensively as a key enzyme involving in the formation of the correct pattern of disulfide bonds in newly synthesized proteins. The relationship between PDI expression and cancer development has attracted interest of cancer researchers in recent years. MATERIALS AND METHODS: We implanted ACI rats with 12 mg Zeranol pellet and harvested the mammary tissues and tumor at day 110 after implantation and investigated the effect of Zeranol-implantation on cell proliferation by histological examination and proliferation in vitro. We also evaluated PDI mRNA expression in primary epithelial cells isolated from normal mammary glands and primary tumor cells from tumor specimens using real-time RT-PCR. To further confirm, we also evaluated the effect of Zeranol on PDI mRNA expression in primary epithelial cells isolated from normal mammary gland of ACI rats. RESULTS: We observed a palpable mammary tumor in one of three Zeranol-implanted ACI rats at day-110 post Zeranol-implantation. Zeranol-implantation significantly promoted the cell proliferation of primary mammary epithelial and stromal cells isolated from the mammary gland of normal ACI rats. PDI mRNA is over-expressed in primary tumor cells isolated from the tumor specimen and in Zeranol-treated primary cultured epithelial cells from the mammary gland of normal ACI rats. CONCLUSION: These findings suggest that up-regulated expression of PDI may play a critical role in mammary tumorigenesis and cell proliferation in response to Zeranol. Our findings implicate PDI as a biomarker for mammary tumorigenesis. PMID- 21617225 TI - Zanthoxylum ailanthoides Sieb and Zucc. extract inhibits growth and induces cell death through G2/M-phase arrest and activation of apoptotic signals in colo 205 human colon adenocarcinoma cells. AB - The effects of 50% ethanolic stem extracts of Zanthoxylum ailanthoides Sieb and Zucc. (ZASZ) on the cell viability, cell cycle and apoptosis were investigated in a human colon adenocarcinoma cell line (colo 205). The results demonstrated that ZASZ induced morphological changes and decreased the cell viability. ZASZ promoted Wee1, checkpoint kinase 2 (CHK2), p21 and p53 levels, decreased cyclin B and cdc25c associated with that led to G(2)/M phase arrest. ZASZ-triggered apoptosis was confirmed by 4' -6-diamidino-2-phenylindole (DAPI) staining and DNA gel electrophoresis. ZASZ increased the levels of glucose-regulated protein 78 (GRP78) and growth arrest and DNA damage inducible gene 153 (GADD153), and promoted an increase of reactive oxygen species (ROS) and Ca(2+) release, and loss of mitochondrial membrane potential (DeltaPsi(m)) accompanied by cytochrome c release that was due to the decrease of Bcl-2 and increase of Bax levels in the colo 205 cells. ZASZ also induced the protein levels of apoptosis-inducing factor (AIF) and endonuclease G (Endo G), increased the levels of caspase-3, -7 and -9, and stimulated the levels of fatty acid synthase (Fas) and Fas ligand in the colo 205 cells. ZASZ contains phenolic compounds, including flavone, chlorogenic acid and isofraxidin, among which, flavone was found to be the most effective in reducing cell viability and proliferative responses in the colo 205 cells. ZASZ induces cytotoxicity and apoptosis in colo 205 cells which provides the rationale for studies in animal models on the utilization of ZASZ as a potential cancer therapeutic compound. PMID- 21617226 TI - Association of FOXP3 expression with non-small cell lung cancer. AB - BACKGROUND/AIM: Lung cancer is rarely cured by current therapeutic approaches. Although numerous studies have implicated FOXP3 positive regulatory T-cells in cancer pathogenesis, the role of FOXP3 in lung cancer pathogenesis remains unkown. MATERIALS AND METHODS: Using immunohistochemistry FOXP3 expression was determined in 44 NSCLC tissue specimens, 20 samples from adjacent non neoplastic lung parenchyma and 5 normal lung tissue specimens. RESULTS: FOXP3 immunostaining was always nuclear in both tumor and non-neoplastic adjacent tissues. FOXP3 was also detected at lower levels in normal bronchial epithelium. Moreover, FOXP3 expression in cancer cells correlated with lymphocytic FOXP3-immunopositivity and the presence of lymph node metastasis. FOXP3 lymphocytic expression was also negatively associated with the age of the patients. CONCLUSION: FOXP3 is overexpressed in NSCLC cells and tumor-infiltrating lymphocytes. This study provides evidence that lymphocytic FOXP3 expression may be age related and that tumor FOXP3 expression is correlated with lymph node metastasis. PMID- 21617227 TI - Gene mutations and polymorphisms of TP53 and FHIT in chronic esophagitis and esophageal carcinoma. AB - AIM: The aim of this study was to investigate genetic changes of the TP53 (tumor protein p53) and FHIT (fragile histidine triad) genes in precursor lesion such as chronic esophagitis (CE) and in esophageal squamous cell carcinoma (ESCC). MATERIALS AND METHODS: PCR-Single Strand Conformation Polymorphism (SSCP) analysis and DNA sequencing in 30 CE and 10 ESCC specimens were performed. RESULTS: DNA sequencing indicated two novel mutations in the TP53 exons 5 (codon 147) and 6 (codon 197) in 2/9 SSCP positive cases of ESCC, but no mutation was found in the CE. The SIFT (Sorting Intolerant From Tolerant) web-based program showed that missense variant at codon 197 of TP53 could affect the protein function. Additionally, polymorphisms of the TP53 exon 4 (codon 36 and 72) and of the FHIT exon 7 (codon 88) were observed in 4/11 (36%) cases of CE and 6/9 (67%) SSCP positive cases of ESCC after DNA sequencing. CONCLUSION: TP53 gene mutations are not common events in CE, but are frequent in ESCC, and as polymorphisms of TP53 and FHIT may confer a greater risk for the development of esophageal carcinoma, further studies are required. PMID- 21617228 TI - Phenethyl isothiocyanate (PEITC) promotes G2/M phase arrest via p53 expression and induces apoptosis through caspase- and mitochondria-dependent signaling pathways in human prostate cancer DU 145 cells. AB - Phenethyl isothiocyanate (PEITC), one of many compounds found in cruciferous vegetables, has been reported as a potential anticancer agent. In earlier studies, PEITC was shown to inhibit cell growth and induction of apoptosis in many cancer cell lines. However, no report has shown whether PEITC can induce apoptosis in human prostate cancer cells. Herein, we aimed to determine whether PEITC has anticancer activity in DU 145 human prostate cancer cells. As a result, we found that PEITC induced a dose-dependent decrease in cell viability through induction of cell apoptosis and cell cycle arrest in the G(2)/M phase of DU 145 cells. PEITC induced morphological changes and DNA damage in DU 145 cells. The induction of G(2)/M phase arrest was mediated by the increase of p53 and WEE1 and it reduced the level of CDC25C protein. The induction of apoptosis was mediated by the activation of caspase-8-, caspase-9- and caspase-3-depedent pathways. Results also showed that PEITC caused mitochondrial dysfunction, increasing the release of cytochrome c and Endo G from mitochondria, and led cell apoptosis through a mitochondria-dependent signaling pathway. This study showed that PEITC might exhibit anticancer activity and become a potent agent for human prostate cancer cells in the future. PMID- 21617229 TI - Potential prognostic value of repulsive guidance molecules in breast cancer. AB - BACKGROUND: Repulsive guidance molecules (RGMs) are novel co-receptors of bone morphogenetic proteins (BMPs) which have been implicated in bone metastasis of cancer. This study aimed to investigate roles played by RGMs in breast cancer. MATERIALS AND METHODS: Expression of RGMs was examined in breast cancer cell lines using RT-PCR. The expression of RGMs in human breast cancer tissues was assessed using both quantitative PCR and immunohistochemical staining. RESULTS: RGMB was detectable in both cell lines and tissues samples of breast cancer. RGMA and RGMC were expressed in the breast tissues, but were undetectable in the examined breast cancer cell lines. Furthermore, reduced expression of RGMA in breast cancer was associated with poor prognosis. RGMB transcript levels appeared to be lower in breast cancer with local recurrence and distant metastasis, but were relatively higher in the patients who died from the disease. CONCLUSION: Aberrant expression of RGMs was indicated in breast cancer. The perturbed expression was associated with disease progression and poor prognosis. PMID- 21617230 TI - Prognostic effects and regulation of activin A, maspin, and the androgen receptor in upper urinary tract urothelial carcinoma. AB - BACKGROUND: Molecular mechanisms responsible for carcinogenesis in upper urinary tract urothelial carcinoma (UUTUC) are not yet clear. This study aimed to examine and correlate the subcellular localization of activin A, maspin, and the androgen receptor (AR) with demographic characteristics, pathological grade, and stage of UUTUC in a Taiwanese population, and to investigate the regulatory mechanisms for activin A, maspin, and AR. MATERIAL AND METHODS: Sections of stage I and II of UUTUCs from 93 patients were examined, with immunohistochemical detection of activin A, maspin, and AR. Patients were divided into four groups according to stage, grade, and disease-free interval (DFI). Pathologic characteristics and the subcellular localization of these markers were correlated with DFI. The urothelial carcinoma cell line HT1197 was stimulated with activin A at different time-points, and the mRNA expression of maspin before and after activin A stimulation was analyzed by reverse transcription-polymerase chain reaction (RT PCR). RESULTS: Expression of AR was observed to be stronger in stage II than in stage I of UUTUC. Expression of cytosolic activin A correlated with longer DFI for early-stage UUTUC (=0.048). Cellular and molecular localization examination revealed that a high level of activin A in the cytosol positively correlated with a high level of maspin in the cytosol (=0.038), and with increased AR expression in the cytosol (=0.044). By RT-PCR, mRNA expression of maspin was significantly induced after administering activin A to HT1197 cancer cells. CONCLUSION: Activin A can induce maspin expression in urothelial carcinoma cells. The expression level and localization of activin A, maspin and AR may be exploited and used as predictive markers for UUTUCs. PMID- 21617231 TI - Response of medulloblastoma cells to vincristine and lomustine: role of TRKC, CTNNB1 and STK15. AB - BACKGROUND: Vincristine and lomustine are two important chemotherapeutic drugs used for the treatment of different types of neoplasms, including medulloblastomas. MATERIALS AND METHODS: We investigated the effects of vincristine and lomustine on 12 primary medulloblastoma cell cultures and the DAOY cell line using the annexinV-flow cytometry and immunoblotting techniques, following treatment of cells for different periods of time. RESULTS: Both drugs triggered apoptosis and cell cycle delay at the G(2)/M phase and also up regulated p16. Furthermore, the expression of 8 different cancer-related genes were assessed and their mRNA and protein levels were found to be highly heterogeneous and did not correlate in several medulloblastoma cultures. Importantly, there was significant correlation between the level of cadherin associated protein beta 1 (CTNNB1) and Aurora kinase A (STK15) proteins and neurotrophic tyrosine kinase receptor type 3 (TRKC) mRNA and the proportion of apoptosis induced by vincristine, the combination of both drugs, and lomustine, respectively. CONCLUSION: These genes could be of great importance as therapeutic biomarkers during the treatment of medulloblastoma patients with vincristine and lomustine. PMID- 21617232 TI - Impact of common medications on serum total prostate-specific antigen levels and risk group assignment in patients with prostate cancer. AB - A recent study in men without prostate cancer suggested that extended use of common medications (nonsteroidal anti-inflammatory drugs (NSAIDs), thiazide diuretics and statins) may lower serum total prostate-specific antigen (PSA) levels by clinically relevant amounts. The present study evaluated the impact of these drugs in patients with clinically localized prostate cancer. A retrospective analysis of 177 patients was performed. The multivariate regression analyses were adjusted for age, prostate volume, Gleason score, T stage, diagnostic setting (clinical symptoms versus elevated PSA only) and presence of diabetes mellitus. Drug use increased with age, e.g. to 50% in patients >=70 years. The most commonly used drugs were statins (32% of all patients, including those who used drug combinations), followed by NSAIDs (21%) and thiazide diuretics (13%). Drug use was associated with a statistically significant PSA reduction (12%, when comparing 104 non-users to 73 users of any of the three drug types; adjusted analysis, p=0.01). Compared to the U.S.A. National Comprehensive Cancer Network risk group assignment based on measured PSA level, reassignment after correcting for medication use resulted in 8 changes among 57 patients with low or intermediate risk (14%). No such changes can be expected in patients belonging to the high-risk group. These results support the concerns expressed previously, given that risk group assignment, which may be inaccurate in patients using concomitant medications, eventually guides choice of treatment. PMID- 21617233 TI - Iron chelation therapy with deferasirox induced complete remission in a patient with chemotherapy-resistant acute monocytic leukemia. AB - BACKGROUND: A patient with chemotherapy-resistant acute monocytic leukemia who achieved complete remission (CR) after iron chelation therapy (ICT) with deferasirox is reported for the first time. A 73-year-old Japanese man with acute monocytic leukemia who was refractory to conventional remission induction chemotherapies achieved a partial response, with some improvement of his hemoglobin level and white blood cell count after gemtuzumab ozogamicin (GO) treatment. Seven months after GO treatment, the disease relapsed and the patient developed pancytopenia. He declined further chemotherapy, and started receiving 1,200-1,800 ml of packed red blood cell transfusion per month together with ICT with deferasirox (baseline serum ferritin level was 1,412 ng/ml). Twelve months after the initiation of deferasirox, the patient's serum ferritin level decreased to below 1,000 ng/ml and deferasirox was discontinued. Four months after discontinuation of deferasirox, the blood cell count normalized and the patient became transfusion-independent. Bone marrow aspiration and biopsy revealed hematological and cytogenetic CR. CONCLUSION: CR was achieved after ICT with deferasirox in a patient with acute myelogenous leukemia, suggesting that deferasirox may have an antileukemic effect in the clinical setting. PMID- 21617234 TI - Treatment of low-risk prostate cancer with radical hypofractionated accelerated radiotherapy with cytoprotection (HypoARC): an interim analysis of toxicity and efficacy. AB - AIM: Radiobiological analysis of clinical data suggests that prostate cancer has a low alpha/beta ratio, implying that large radiotherapy fractions may better control the disease. Acceleration of radiotherapy may be also of importance in a subset of tumors. In this study we assessed the feasibility and efficacy of a highly accelerated and hypofractionated scheme of radiotherapy (HypoARC), for the treatment of localized low risk prostate cancer. PATIENTS AND METHODS: Fifty-five patients with prostate cancer (T1-2 stage, Gleason score <7 and prostate specific antigen (PSA) <10 ng/ml) were treated with localized conformal 4-field radiotherapy to the prostate and seminal vesicles: 51 Gy were delivered (3.4 Gy/fraction, within 19 days). The biological dose to the prostate ranged from 67.9-91.7 Gy. Amifostine (0-1000 mg depending upon tolerance) was delivered daily for cytoprotection. The median follow-up of patients is 30 (6-69) months. RESULTS: Early toxicity was overall low, proctitis being the most frequent side effect (23.6% grade II). High dose amifostine significantly protected against proctitis (p=0.005). Grade 2 frequency and dysurea occurred in 1.8% and 3.7% of cases, respectively. There was no late toxicity >=grade 2. Amifostine significantly protected against chronic frequency (p=0.02). Within a median follow-up of 30 months, one patient (1.8%) experienced a biochemical relapse. CONCLUSION: HypoARC is feasible and safe for patients with low-risk prostate cancer and, considering also the high efficacy noted, a strong rationale is provided for the further evaluation of HypoARC in randomized trials. PMID- 21617235 TI - Pemetrexed-induced hyperpigmentation of the skin. AB - Pemetrexed (AlimtaTM) is used frequently for the treatment of lung cancer and is associated with various types and grades of cutaneous side-effects. We report here one patient who presented with asymptomatic hyperpigmentation of the skin, localized on the palms of the hands and the soles of the feet after administration of pemetrexed for lung cancer despite receiving standard pre medication. This type of skin toxicity was completely reversible after withdrawal of the drug (without pharmacologic intervention) and has to our knowledge not been reported before. PMID- 21617236 TI - Lymphatic microvessel density, VEGF-C, and VEGFR-3 expression in different molecular types of breast cancer. AB - BACKGROUND: Breast cancer is a heterogeneous disease and five major distinct molecular types have been characterized by gene analysis and immunohistochemistry. The molecular types of breast cancer have different behavior, a particular profile of response to therapy, reflected in the differential survival of patients. Previous findings showed a particular preference for lymph node and distant metastases of different molecular types, but the specific lymphangiogenic profile of these types is lacking. PATIENTS AND METHODS: We investigated the differential expression vascular endothelial growth factor-C (VEGF-C), vascular endothelial growth factor receptor-3 (VEGFR-3) and D2 40 by immunohistochemistry, to evaluate lymphangiogenesis and the lymphatic microvessel density (LMVD), in patients with breast cancer, stratified according to the molecular classification. RESULTS: There was a differential expression of VEGF-C/VEGFR-3 and D2-40 in different molecular types of breast cancer, with highest level of expression for these markers being found in HER2 and luminal B types and the lowest in basal-like type. The lowest value of both intratumoral and peritumoral LMVD were found in normal-like type breast cancer. VEGF-C expression did not correlate with the grade of the tumor, but a significant correlation was found with lymph node metastasis. VEGFR-3 expression was found in 66.66% of the cases and correlated with the expression of VEGF-C in tumor cells. There was a positive correlation between VEGF-C, VEGFR-3 and LMVD only in the HER2 type, and a positive correlation in HER2 and normal-like types with VEGFR-3 expression in tumor cells. In addition, there was a correlation between HER2 type, VEGF-C and VEGFR-3 expression in tumor cells and lymphatic endothelium, respectively, and LMVD. CONCLUSION: Our results support a differential signature of lymphangiogenesis in different molecular types of breast cancer and these findings may have a direct impact on prognosis and therapeutic strategy of this disease. PMID- 21617237 TI - Off-label use of oxaliplatin in patients with metastatic breast cancer. AB - BACKGROUND: Oxaliplatin is an anticancer agent only approved for treatment of colorectal cancer, but that has shown some activity in metastatic breast cancer in phase II studies. Herein, we examine the off-label use of oxaliplatin in unselected patients with metastatic breast cancer. PATIENTS AND METHODS: A retrospective review was performed of all patients with metastatic breast cancer treated with oxaliplatin at our hospital between February 2003 and November 2009. Data concerning patterns of use, safety and activity were collected from patient charts. RESULTS: The cohort comprised 30 female patients with a median age of 49 (range, 34-68 years) and a median of two involved organs (range, 1-4). All patients had been pretreated for metastatic breast cancer (median number of previous lines: 3; range:1-6). Oxaliplatin was only given in association either with fluorouracil and folinic acid (n=23) or with gemcitabine (n=7). The most commonly used dose was 100 mg/m(2) given every other week or every 3 weeks. As of December 15, 2009, the median duration of treatment was 4 (range, 0.75-11) months. Most of the discontinuations occured due to disease progression (n=11) and adverse effects or worsening condition (n=8). Twelve (40%) patients presented side-effects related to oxaliplatin use including hematotoxicity (n=8), gastrointestinal disorders (n=4) and neuropathies (n=2). Among patients evaluable for antitumoral activity (n=15), one patient achieved a complete response and one patient demonstrated a partial response. Most of the patients (57%) continued to be treated by chemotherapy after oxaliplatin. Median overall survival for the evaluable patients was 10 (range, 1-51) months. CONCLUSION: In our population of heavily pretreated women with metastatic breast cancer, off-label use of oxaliplatin was of little worth. This off-label treatment was not the last therapeutic option for most of these patients. PMID- 21617238 TI - Correlation between tumor-associated proteins and response to neoadjuvant treatment in patients with advanced squamous-cell esophageal cancer. AB - BACKGROUND: Possible predictive markers of response to neoadjuvant radiochemotherapy (NRCT) of esophageal cancer have been identified. PATIENTS AND METHODS: Patient biopsies were obtained from both tumor and normal tissue before the NRCT of locally advanced esophageal squamous cell carcinoma. Protein solutions were separated and immunoblot analysis was performed with heat shock protein (Hsp)16.2, heme-binding protein 2 (SOUL), BCL2-associated X protein (Bax), B-cell-associated leukemia protein 2 (Bcl-2) and heat shock protein 90 (Hsp90) antibodies. Following NRCT, the patients were restaged according to the Response Evaluation Criteria In Solid Tumors (RECIST). Following resections the pathological down-staging was evaluated. RESULTS: Clinical restaging revealed a response rate of 65%. Pathological examination revealed down-staging in 30% and 25% of the cases for the T and N categories respectively. Compared to the normal esophageal mucosa, a decreased expression of Hsp16.2, Hsp90 and SOUL proteins and an increased Bax/Bcl-2 ratio was found in the responding tumors. CONCLUSION: Hsp16,2, Hsp90 and SOUL expression and Bax/ Bcl-2 ratio correlates to the efficacy of NRCT and predict outcome in patients with locally advanced squamous cell esophageal cancer. PMID- 21617239 TI - Bevacizumab in first-line therapy of metastatic colorectal cancer: a retrospective comparison of FOLFIRI and XELIRI. AB - BACKGROUND: The antivascular endothelial growth factor monoclonal antibody bevacizumab with infusional 5-fluorouracil, leucovorin and irinotecan (FOLFIRI) is a standard first-line treatment option for metastatic colorectal cancer. However, clinical data for capecitabine and irinotecan (XELIRI) with bevacizumab are limited. PATIENTS AND METHODS: A retrospective study was conducted on 139 patients with metastatic colorectal cancer to assess the efficacy and safety of first-line bevacizumab in combination with XELIRI or FOLFIRI. Primary endpoints were overall response rate (ORR), disease control rate and radical resection rate. Secondary endpoints included overall survival (OS), progression-free survival (PFS) and safety. RESULTS: No significant differences in efficacy were observed between patients administered XELIRI or FOLFIRI with bevacizumab. The ORR, median OS and PFS and recorded adverse events (AEs) were comparable to those previously reported, with no new or unexpected AEs observed. CONCLUSION: Bevacizumab is similarly efficacious and well tolerated when administered with XELIRI or FOLFIRI. PMID- 21617240 TI - Differences in breast carcinoma in situ between menopausal and premenopausal women. AB - AIM: To seek differences between ductal carcinoma in situ (DCIS) according to the menopausal status of patients and to analyze their repercussions on patient care. PATIENTS AND METHODS: A multicenter retrospective study of 384 patients from 3 centers specialized in breast cancer surgery was carried out based on an analysis of the various characteristics (clinical, therapeutic, histologic, outcome) of DCIS between two groups of post- and pre-menopausal patients. RESULTS: At the time of diagnosis, 58.6% of the patients were menopausal. Compared to these patients, DCIS in premenopausal women was more frequently associated with initial clinical signs (p=0.006), a larger tumor size (p=0.02), involved margins after initial surgery (p=0.005), and surgical re-excision (p=0.03). The mammograms of the menopausal patients indicated a worse prognosis (using the American College of Radiology Classification) (p=0.025), and according to the histology report findings, more marked comedo necrosis (p=0.01). There was no difference in the other criteria (nuclear grade, multifocality, benign lesions associated with malignancy, relapse and its time of occurrence). The use of hormone replacement therapy had no effect on these data. CONCLUSION: The characteristics of DCIS are similar, whether occuring before or after the menopause, but the phenotypic expression is different. Menopausal status should not be a criterion for changing patient care. PMID- 21617241 TI - Fundic gland polyps. AB - BACKGROUND: Fundic gland polyps (FGPs) are common hamartomatous cystic lesions in the gastric mucosa. The aim was to assess the cellular components in sections stained with Giemsa or toluidine blue: two stains that easily differentiate parietal from chief cells. MATERIALS AND METHODS: Sixty-eight consecutive gastric biopsies having well-oriented sections from the fundic mucosa were investigated: 34 had FGP, and 34 were control cases. Using an ocular microscale, the thickness of each FGP was divided into an upper and a lower compartment. The proportion of parietal and chief cells in each compartment was assessed in 3 consecutive high power fields (*40). RESULTS: In FGPs, parietal cells predominated in the upper compartment where the parietal chief cell ratio >=60 was 59.8%, and in the lower compartment it was only 5.9% (p<0.05). Within cysts, cell exfoliation with secondary eosinophilic nuclear-free material was found. FGPs also displayed small glandular islands with an admixture of parietal and chief cells. The cytoplasm of parietal cells was often ballooned, vacuolated, with basophilic deposits or anucleated (ghost cells). CONCLUSION: FGPs are composed of parietal cells predominantly in the upper half and with chief cells in the lower half, as well as with glandular islands showing cytological alterations in parietal cells. Intraluminal nuclear-free material seems to obstruct the outlets of the glands leading to cystic formation. PMID- 21617242 TI - Current concepts in the non-operative management of rectal cancer after neoadjuvant chemoradiation. AB - BACKGROUND: Neoadjuvant chemoradiation (CRT) is used to downstage locally advanced rectal cancer prior to surgery. This is an established paradigm of treatment in the West. Some patients who undergo chemoradiotherapy achieve a complete clinical response, where no evidence of tumour can be found on clinical assessment prior to surgery. Recent studies have suggested a 'watch and wait policy' for patients who achieve a complete clinical response with close clinical follow-up, and to only intervene surgically if local recurrence manifests. This notion differs from established regimes of treatment, but by virtue of its positive implications for patients, deserves further consideration and scrutiny. This article reviews the evidence for the non-operative 'watch and wait' management of rectal cancer in patients with complete clinical response after chemoradiation, and discusses the limitations in applying a policy of this nature. METHODS: The evidence in the literature for non-operative treatment of. Only a few studies have examined this issue. Results have been variable, with only one study supporting non-operative management, where low local recurrence rates were shown on intermediate follow-up (<3%), with excellent overall and disease-free survival rates (100% and 86% respectively). Another two studies found high local recurrence rates (approximately 80%) and poor survival on short term follow-up (around 20%). Differences in selection of patients were found between studies, as well as other aspects that might explain some of the disparity in results found. CONCLUSION: There is not enough evidence to justify a non-operative approach to treating rectal cancer after chemoradiation in complete clinical responders at present. Better designed studies are required to clarify some of the issues involved. PMID- 21617243 TI - Assessment of general anxiety in patients with breast disease and breast cancer using the Spielberger STAI self evaluation test: a prospective case-control study in Finland. AB - BACKGROUND: Trait anxiety (T-Anxiety) was assessed using the subscale from the Spielberger State-Trait Anxiety Inventory test (STAI form Y-2), with statements refering to how a person generally feels, and a higher total score reflecting a higher T-Anxiety. To the authors' knowledge, the associations between the STAI test and the risk of breast cancer (BC) are rarely considered together in a prospective study. PATIENTS AND METHODS: In an extension of the Kuopio Breast Cancer Study 115 women with breast symptoms were evaluated for anxiety using the STAI form Y-2 test before any diagnostic procedures were carried out. RESULTS: The clinical examination and biopsy showed BC in 34 patients, benign breast disease (BBD) in 53 patients and 28 individuals were shown to be healthy study subjects (HSS). There was a trend for the HSS women to have less severe anxiety in the STAI test (1/28 patients, 3.6%) than those in the BBD (7/53 patients, 13.2%) and BC groups (4/34 patients, 11.8%). The subjects in the HSS group reported being significantly more happy in the STAI test (STAI mean score, 1.54) than the BBD group (STAI mean score, 1.98) and the BC group (STAI mean score, 2.00) (p=0.01). The subjects in the HSS group also reported being more content in the STAI test (STAI mean score, 1.68) than the BBD group (STAI mean score, 1.87) and the BC group (STAI mean score, 1.97). CONCLUSION: Patients with BC and BBD tend to have more unhappy and uncontent feelings, but no specific link between the STAI test (form Y-2) and breast cancer risk is supported. PMID- 21617246 TI - Separating foliar physiology from morphology reveals the relative roles of vertically structured transpiration factors within red maple crowns and limitations of larger scale models. AB - A spatially explicit mechanistic model, MAESTRA, was used to separate key parameters affecting transpiration to provide insights into the most influential parameters for accurate predictions of within-crown and within-canopy transpiration. Once validated among Acer rubrum L. genotypes, model responses to different parameterization scenarios were scaled up to stand transpiration (expressed per unit leaf area) to assess how transpiration might be affected by the spatial distribution of foliage properties. For example, when physiological differences were accounted for, differences in leaf width among A. rubrum L. genotypes resulted in a 25% difference in transpiration. An in silico within canopy sensitivity analysis was conducted over the range of genotype parameter variation observed and under different climate forcing conditions. The analysis revealed that seven of 16 leaf traits had a >=5% impact on transpiration predictions. Under sparse foliage conditions, comparisons of the present findings with previous studies were in agreement that parameters such as the maximum Rubisco-limited rate of photosynthesis can explain ~20% of the variability in predicted transpiration. However, the spatial analysis shows how such parameters can decrease or change in importance below the uppermost canopy layer. Alternatively, model sensitivity to leaf width and minimum stomatal conductance was continuous along a vertical canopy depth profile. Foremost, transpiration sensitivity to an observed range of morphological and physiological parameters is examined and the spatial sensitivity of transpiration model predictions to vertical variations in microclimate and foliage density is identified to reduce the uncertainty of current transpiration predictions. PMID- 21617247 TI - Brassinosteroids can regulate cellulose biosynthesis by controlling the expression of CESA genes in Arabidopsis. AB - The phytohormones, brassinosteroids (BRs), play important roles in regulating cell elongation and cell size, and BR-related mutants in Arabidopsis display significant dwarf phenotypes. Cellulose is a biopolymer which has a major contribution to cell wall formation during cell expansion and elongation. However, whether BRs regulate cellulose synthesis, and if so, what the underlying mechanism of cell elongation induced by BRs is, is unknown. The content of cellulose and the expression levels of the cellulose synthase genes (CESAs) was measured in BR-related mutants and their wild-type counterpart. The chromatin immunoprecipitation (CHIP) experiments and genetic analysis were used to demonstrate that BRs regulate CESA genes. It was found here that the BR-deficient or BR-perceptional mutants contain less cellulose than the wild type. The expression of CESA genes, especially those related to primary cell wall synthesis, was reduced in det2-1 and bri1-301, and was only inducible by BRs in the BR-deficient mutant det2-1. CHIP experiments show that the BR-activated transcription factor BES1 can associate with upstream elements of most CESA genes particularly those related with the primary cell wall. Furthermore, over expression of the BR receptor BRI1 in CESA1, 3, and 6 mutants can only partially rescue the dwarf phenotypes. Our findings provide potential insights into the mechanism that BRs regulate cellulose synthesis to accomplish the cell elongation process in plant development. PMID- 21617248 TI - Dynamic trafficking of wheat gamma-gliadin and of its structural domains in tobacco cells, studied with fluorescent protein fusions. AB - Prolamins, the main storage proteins of wheat seeds, are synthesized and retained in the endoplasmic reticulum (ER) of the endosperm cells, where they accumulate in protein bodies (PBs) and are then exported to the storage vacuole. The mechanisms leading to these events are unresolved. To investigate this unconventional trafficking pathway, wheat gamma-gliadin and its isolated repeated N-terminal and cysteine-rich C-terminal domains were fused to fluorescent proteins and expressed in tobacco leaf epidermal cells. The results indicated that gamma-gliadin and both isolated domains were able to be retained and accumulated as protein body-like structures (PBLS) in the ER, suggesting that tandem repeats are not the only sequence involved in gamma-gliadin ER retention and PBLS formation. The high actin-dependent mobility of gamma-gliadin PBLS is also reported, and it is demonstrated that most of them do not co-localize with Golgi body or pre-vacuolar compartment markers. Both gamma-gliadin domains are found in the same PBLS when co-expressed, which is most probably due to their ability to interact with each other, as indicated by the yeast two-hybrid and FRET-FLIM experiments. Moreover, when stably expressed in BY-2 cells, green fluorescent protein (GFP) fusions to gamma-gliadin and its isolated domains were retained in the ER for several days before being exported to the vacuole in a Golgi-dependent manner, and degraded, leading to the release of the GFP 'core'. Taken together, the results show that tobacco cells are a convenient model to study the atypical wheat prolamin trafficking with fluorescent protein fusions. PMID- 21617249 TI - An epigenetic view of plant cells cultured in vitro: somaclonal variation and beyond. AB - Epigenetic mechanisms are highly dynamic events that modulate gene expression. As more accurate and powerful tools for epigenetic analysis become available for application in a broader range of plant species, analysis of the epigenetic landscape of plant cell cultures may turn out to be crucial for understanding variant phenotypes. In vitro plant cell and tissue culture methodologies are important for many ongoing plant propagation and breeding programmes as well as for cutting-edge research in several plant model species. Although it has long been known that in vitro conditions induce variation at several levels, most studies using such conditions rely on the assumption that in vitro cultured plant cells/tissues mostly conform genotypically and phenotypically. However, when large-scale clonal propagation is the aim, there has been a concern in confirming true-to-typeness using molecular markers for evaluating stability. While in most reports genetic variation has been found to occur at relatively modest frequencies, variation in DNA methylation patterns seems to be much more frequent and in some cases it has been directly implicated in phenotypic variation. Recent advances in the field of epigenetics have uncovered highly dynamic mechanisms of chromatin remodelling occurring during cell dedifferentiation and differentiation processes on which in vitro adventitious plant regeneration systems are based. Here, an overview of recent findings related to developmental switches occurring during in vitro culture is presented. Additionally, an update on the detection of epigenetic variation in plant cell cultures will be provided and discussed in the light of recent progress in the plant epigenetics field. PMID- 21617251 TI - Endometrial stromal cells and immune cell populations within lymph nodes in a nonhuman primate model of endometriosis. AB - Mounting evidence suggests that immunological responses may be altered in endometriosis. The baboon (Papio anubis) is generally considered the best model of endometriosis pathogenesis. The objective of the current study was to investigate for the first time immunological changes within uterine and peritoneal draining lymph nodes in a nonhuman primate baboon model of endometriosis. Paraffin-embedded femoral lymph nodes were obtained from 22 normally cycling female baboons (induced endometriosis n = 11; control n = 11). Immunohistochemical staining was performed with antibodies for endometrial stromal cells, T cells, immature and mature dendritic cells, and B cells. Lymph nodes were evaluated using an automated cellular imaging system. Endometrial stromal cells were significantly increased in lymph nodes from animals with induced endometriosis, compared to control animals (P = .033). In animals with induced endometriosis, some lymph node immune cell populations including T cells, dendritic cells and B cells were increased, suggesting an efficient early response or peritoneal drainage. PMID- 21617252 TI - Developmental and dyadic perspectives on commitment in adult romantic relationships. AB - We tested hypotheses concerning the developmental roots of becoming the "weak link" (less committed) partner in adult romantic relationships and the associations between partners' absolute and relative levels of commitment and dyadic outcomes. We examined 78 target 20- to 21-year-olds who were involved in a romantic relationship and who had been studied since birth. As predicted, people who received lower-quality support from caregivers in toddlerhood or who were less able to resolve conflicts with a best friend in midadolescence were more likely to become the weak-link partner in a romantic relationship at age 20 to 21. Furthermore, lower commitment on the part of the weak-link partner coupled with greater discrepancy in commitment between partners predicted a greater likelihood that the couple would display hostility (rated by observers) during a videotaped conflict-resolution task when they were 20 to 21 years old. These findings are discussed from developmental and dyadic perspectives. PMID- 21617253 TI - Pain perception in knees with circumscribed cartilage lesions is associated with intra-articular IGF-1 expression. AB - BACKGROUND: Circumscribed cartilage defects are considered as prearthritic lesions and lead to differential intra-articular cytokine expression. Mechanisms of associated pain development and influence of smoking behavior are not yet fully understood in humans. PURPOSE: This study aimed to reveal relations between synovial cytokine levels in knees with circumscribed cartilage defects and pain sensation. STUDY DESIGN: Descriptive laboratory study. METHODS: In a clinical trial, knee lavage fluids of 42 patients with circumscribed cartilage lesions treated by either microfracturing (n = 19) or by autologous chondrocyte implantation (n = 23) and fluids of 5 healthy control individuals were prospectively collected. Preoperative knee pain was evaluated according to frequency and strength; subjective knee function was assessed using a visual analog scale and the International Knee Documentation Committee (IKDC) score. Synovial concentrations of aggrecan, insulin-like growth factor (IGF)-I, basic fibroblast growth factor (bFGF), interleukin (IL)-1beta, bone morphogenetic protein (BMP)-2, and BMP-7 were determined by enzyme-linked immunosorbent assay. RESULTS: Pain strength showed a highly significant association with intra articular IGF-1 levels (rho = .48, P < .01), but no correlation with synovial concentrations of aggrecan, bFGF, IL-1beta, BMP-2, and BMP-7. Although pain strength and frequency demonstrated a statistically significant relationship, no substantial association between pain frequency and any of the examined cytokine levels was found. Intra-articular IGF-1 concentrations significantly correlated with the area of cartilage damage (rho = .35, P < .02); the other investigated cytokines failed to show this association. Neither of the determined intra articular mediators demonstrated statistically significant correlations with subjective knee function or IKDC score. Only intra-articular concentrations of IGF-1 and BMP-2 statistically significantly correlated with age; total protein content was negatively associated with body mass index (P < .05). In smokers, synovial expression of total protein content, IGF-1, and bFGF was significantly diminished compared to nonsmokers (P < .05). CONCLUSION: Insulin-like growth factor-I is present in knees with circumscribed cartilage lesions in a size dependent manner. IGF-1 levels correlated with indicators of pain perception; smoking negatively influenced synovial cytokine expression related to cartilage metabolism, but pain perception was not altered. PMID- 21617254 TI - Accuracy of palpation-guided and ultrasound-guided needle tip placement into the deep and superficial posterior leg compartments. AB - BACKGROUND: Invasive leg compartment testing may be performed to diagnose chronic exertional compartment syndrome. No prior study has assessed the accuracy of leg compartment needle tip access. PURPOSE: This study was undertaken to describe and determine the accuracy of palpation-guided and ultrasound-guided techniques for needle tip placement into the deep and superficial posterior leg compartments by a less experienced clinician and a more experienced clinician using a cadaveric model. STUDY DESIGN: Controlled laboratory study. METHODS: Twenty unembalmed adult lower limb cadaveric specimens were used for the study. Two investigators, a sports medicine fellow with 10 months of musculoskeletal ultrasound experience and a staff physiatrist with 3 years of musculoskeletal ultrasound experience, each performed 1 palpation-guided and 1 ultrasound-guided colored latex dye injection into the deep and superficial posterior leg compartments of each cadaveric specimen. A blinded investigator dissected the specimens and graded them for accuracy. RESULTS: The accuracy rates of palpation-guided (accuracy rate, 90%; 95% confidence interval [CI], 76%-97%) and ultrasound-guided (accuracy rate, 88%; 95% CI, 73%-95%) deep posterior compartment injections were statistically equivalent (P = 1.000). All 80 injections performed into the superficial posterior compartment were accurate (accuracy rate, 100%; 95% CI, 89% 100%). The accuracy of the less experienced investigator (total injection accuracy rate, 88%; 95% CI, 73%-95%) and the more experienced investigator (total injection accuracy rate, 90%; 95% CI, 76%-97%) were not significantly different (P = 1.000). CONCLUSION: Needle tip placement into the deep and superficial posterior leg compartments is relatively accurate with palpation guidance regardless of level of experience, and does not improve with the use of ultrasound guidance. CLINICAL RELEVANCE: Ultrasound guidance does not appear to be indicated for routine deep or superficial posterior leg compartment pressure testing. However, this does not preclude the need for ultrasound guidance in selected clinical scenarios. PMID- 21617255 TI - Microtriplication of 11q24.1: a highly recognisable phenotype with short stature, distinctive facial features, keratoconus, overweight, and intellectual disability. AB - BACKGROUND: Partial tetrasomy is mainly described as a cytogenetically visible rearrangement due to a supernumerary chromosome (i(12p), i(18p), inv dup(15)). Except for chromosome 15q11q13, intrachromosomal triplications are rare and so far not associated with a recognisable phenotype. METHODS AND RESULTS: This report describes two unrelated patients with a de novo non-recurrent submicroscopic interstitial triplication 11q24.1 detected with array comparative genomic hybridisation and confirmed by fluorescence in situ hybridisation, molecular combing, and quantitative PCR. Microsatellite analysis suggested that a common mechanism of rearrangement might have been involved. These patients share remarkably similar clinical features including distinctive facial dysmorphisms, short stature with small extremities, keratoconus, overweight, and intellectual disability. The overlapping region of 1.8 Mb contains 11 RefSeq genes and three microRNA related genes. Interestingly, the overexpression of ASAM, a gene encoding an adipocyte specific adhesion molecule, may contribute to patients' obesity. Upregulation of BILD, known to mediate apoptosis in a caspase dependent manner, could deserve further investigation into the pathological mechanism of keratoconus. CONCLUSION: Isolated duplications of distal 11q region have been previously reported and associated with intellectual disability but without a consistent set of clinical features. These findings support the proposal that microtriplication 11q24.1 is a well recognisable clinical entity. PMID- 21617250 TI - Comparative transcriptomics of Eastern African cichlid fishes shows signs of positive selection and a large contribution of untranslated regions to genetic diversity. AB - The hundreds of endemic species of cichlid fishes in the East African Great Lakes Tanganyika, Malawi, and Victoria are a prime model system in evolutionary biology. With five genomes currently being sequenced, eastern African cichlids also represent a forthcoming genomic model for evolutionary studies of genotype to-phenotype processes in adaptive radiations. Here we report the functional annotation and comparative analyses of transcriptome data sets for two eastern African cichlid species, Astatotilapia burtoni and Ophthalmotilapia ventralis, representatives of the modern haplochromines and ectodines, respectively. Nearly 647,000 expressed sequence tags were assembled in more than 46,000 contigs for each species using the 454 sequencing technology, largely expanding the current sequence data set publicly available for these cichlids. Total predicted coverage of their proteome diversity is approximately 50% for both species. Comparative qualitative and quantitative analyses show very similar transcriptome data for the two species in terms of both functional annotation and relative abundance of gene ontology terms expressed. Average genetic distance between species is 1.75% when all transcript types are considered including nonannotated sequences, 1.33% for annotated sequences only including untranslated regions, and decreases to nearly half, 0.95%, for coding sequences only, suggesting a large contribution of noncoding regions to their genetic diversity. Comparative analyses across the two species, tilapia and the outgroup medaka based on an overlapping data set of 1,216 genes (~526 kb) demonstrate cichlid-specific signature of disruptive selection and provide a set of candidate genes that are putatively under positive selection. Overall, these data sets offer the genetic platform for future comparative analyses in light of the upcoming genomes for this taxonomic group. PMID- 21617257 TI - Respiratory chain complex I deficiency due to NDUFA12 mutations as a new cause of Leigh syndrome. AB - BACKGROUND: This study investigated a girl with Leigh syndrome born to first cousin parents of Pakistani descent with an isolated respiratory chain complex I deficiency in muscle and fibroblasts. Her early development was delayed, and from age 2 years she started losing motor abilities. Cerebral MRI showed basal ganglia lesions typical of Leigh syndrome. METHODS AND RESULTS: A genome-wide search for homozygosity was performed with the Affymetrix GeneChip 50K Xba array. The analysis revealed several homozygous regions. Three candidate genes were identified, and in one of the genes, NDUFA12, a homozygous c.178C->T mutation leading to a premature stop codon (p.Arg60X) was found. Western blot analysis showed absence of NDUFA12 protein in patient fibroblasts and functional complementation by a baculovirus system showed restoration of complex I activity. CONCLUSION: NDUFA12 mutations are apparently not a frequent cause of complex I deficiency, since mutations were not found by screening altogether 122 complex I deficient patients in two different studies. NDUFA12 encodes an accessory subunit of complex I and is a paralogue of NDUFAF2. Despite the complete absence of NDUFA12 protein, a fully assembled and enzymatically active complex I could be found, albeit in reduced amounts. This suggests that NDUFA12 is required either at a late step in the assembly of complex I, or in the stability of complex I. PMID- 21617256 TI - Variants in or near KITLG, BAK1, DMRT1, and TERT-CLPTM1L predispose to familial testicular germ cell tumour. AB - BACKGROUND: Familial testicular germ cell tumours (TGCTs) and bilateral TGCTs comprise 1-2% and 5% of all TGCTs, respectively, but their genetic basis remains largely unknown. AIM: To investigate the contribution of known testicular cancer risk variants in familial and bilateral TGCTs. METHODS AND RESULTS: The study genotyped 106 single nucleotide polymorphisms (SNPs) in four regions (BAK1, DMRT1, KITLG, TERT-CLPTM1L) previously identified from genome-wide association studies of TGCT, including risk single nucleotide polymorphisms (SNPs) rs210138 (BAK1), rs755383 (DMRT1), rs4635969 (TERT-CLPTM1L) in 97 cases with familial TGCT and 22 affected individuals with sporadic bilateral TGCT as well as 871 controls. Using a generalised estimating equations method that takes into account blood relationships among cases, the associations with familial and bilateral TGCT were analysed. Three previously identified risk SNPs were found to be associated with familial and bilateral TGCT (rs210138: OR 1.80, CI 1.35 to 2.41, p=7.03*10(-5); rs755383: OR 1.67, CI 1.23 to 2.22, p=6.70*10(-4); rs4635969: OR 1.59, CI 1.16 to 2.19, p=4.07*10(-3)). Evidence for a second independent association was found for an SNP in TERT (rs4975605: OR 1.68, CI 1.23 to 2.29, p=1.24*10(-3)). Another association with an SNP was identified in KITLG (rs2046971: OR 2.33, p=1.28*10( 3)); this SNP is in high linkage disequilibrium (LD) with reported risk variant rs995030. CONCLUSION: This study provides evidence for replication of recent genome-wide association studies results and shows that variants in or near BAK1, DMRT1, TERT-CLPTM1L, and KITLG predispose to familial and bilateral TGCT. These findings imply that familial TGCT and sporadic TGCT share a common genetic basis. PMID- 21617258 TI - Array comparative genomic hybridisation on first polar bodies suggests that non disjunction is not the predominant mechanism leading to aneuploidy in humans. AB - INTRODUCTION: Aneuploidy (the presence of extra or missing chromosomes) arises primarily through chromosome segregation errors in the oocyte at meiosis I but the details of mechanism by which such errors occur in humans are the subject of some debate. It is generally believed that aneuploidy arises primarily as a result of segregation of a whole chromosome to the same pole as its homologue (non-disjunction). Nonetheless, classical cytogenetic studies suggest that this model does not fully account for the patterns observed in human oocytes. An alternative model (precocious separation of sister chromatids) has thus been proposed, but recurring criticism of this model purports that technical issues may have led to interpretation errors. MATERIALS AND METHODS: Array comparative genomic hybridisation (aCGH) was used on 164 human first polar bodies to distinguish between whole chromosome (non-disjunction) and chromatid (precocious separation) errors. RESULTS: Single chromatid errors were over 11 times more common than whole chromosome errors, consistent with prior classical cytogenetic and fluorescence in situ hybridisation (FISH) studies. DISCUSSION: The received wisdom that non-disjunction is the primary mechanism leading to human aneuploidy should be reconsidered. PMID- 21617259 TI - New approaches to estimating national rates of invasive pneumococcal disease. AB - National infectious disease incidence rates are often estimated by standardizing locally derived rates using national-level age and race distributions. Data on other factors potentially associated with incidence are often not available in the form of patient-level covariates. Including characteristics of patients' area of residence may improve the accuracy of national estimates. The authors used data from the Centers for Disease Control and Prevention's Active Bacterial Core Surveillance program (2004-2005), adjusted for census-based variables, to estimate the national incidence of invasive pneumococcal disease (IPD). The authors tested Poisson and negative binomial models in a cross-validation procedure to select variables best predicting the incidence of IPD in each county. Including census-level information on race and educational attainment improved the fit of both Poisson and negative binomial models beyond that achieved by adjusting for other census variables or by adjusting for an individual's race and age alone. The Poisson model with census-based predictors led to a national estimate of IPD of 16.0 cases per 100,000 persons as compared with 13.5 per 100,000 persons using an individual's age and race alone. Accuracy of, and confidence intervals for, these estimates can only be determined by obtaining data from other randomly selected US counties. However, incorporating census-derived characteristics should be considered when estimating national incidence of IPD and other diseases. PMID- 21617260 TI - Intergenerational correlations in size at birth and the contribution of environmental factors: The Uppsala Birth Cohort Multigenerational Study, Sweden, 1915-2002. AB - Sizes at birth of parents and their children are known to be correlated, reflecting in part the influence of fetal and maternal genes. Sociodemographic factors, regarded as aspects of the shared environment across generations, would also be expected to contribute, but evidence is limited. In the present study, the authors aimed to quantify the role of the shared environment in explaining intergenerational correlations in birth weight and length by using data across 3 consecutive generations from the Uppsala Birth Cohort Multigenerational Study in Uppsala, Sweden. That study included birth and sociodemographic data on 7,657 singletons born in Uppsala in 1915-1929 (generation 1) and their grandchildren (generation 3). Standard regression and biometric models were used to study the correlations in size at birth of generation 1-generation 3 pairs. The data showed stronger correlations in maternal pairs than in paternal pairs for birth weight (0.125 vs. 0.096, P = 0.02) but not for birth length (0.097 vs. 0.093, P = 0.77). These correlations were not reduced by adjustment for sociodemographic factors in regression models. In contrast, significant shared-environment contributions to the intergenerational correlations were identified in biometric models, averaging 14% for both birth measures. These models assumed a common latent factor for the sociodemographic variables. The present results show that the shared environment moderately but significantly contributes to intergenerational correlations. PMID- 21617261 TI - Incidence of hospital-acquired pressure ulcers in acute care using two different risk assessment scales: results of a retrospective study. AB - More than one million people develop pressure ulcers (PU) every year in the US - a major problem that leads to increased morbidity and mortality. A hospital acquired PU (HAPU) is defined as any ulcer noted 24 or more hours after hospital admission. The purpose of this retrospective study was to compare the incidence of HAPUs between two 1-year periods of time (March 1, 2007 through March 17, 2008 and March 18, 2008 through March 31, 2009) at an acute care facility. Records from all admitted adult medical-surgical and critical care patients along with routinely submitted quality control reports on patients who acquired PUs during hospitalization were reviewed and abstracted. A locally developed PU risk assessment instrument (SST) was used during the first time period and the Braden Scale (BS) was used during the second time period. The overall PU incidence was 368 in 41,840 patient admissions (8.80 per 1,000); 9.49 in the SST and 8.08 in the BS patient sample (P = 0.125). No significant differences in patient age, race, and PU severity or location were observed. However, the incidence of more severe ulcers (Stage III and Stage IV) was lower in the BS group (0.5667 in the SST group compared to 0.2419 per 1,000 admissions in the BS group) even though median patient length of stay was longer (28 days and 34 days in the SST and BS groups, respectively), suggesting that use of a valid and reliable PU risk assessment instrument may reduce the incidence of severe PUs. Study design and sample size limit the ability to generalize the results of this study. Controlled clinical studies comparing the effect of different PU risk assessment instruments on PU incidence are needed. PMID- 21617262 TI - Using chamomile solution or a 1% topical hydrocortisone ointment in the management of peristomal skin lesions in colostomy patients: results of a controlled clinical study. AB - Peristomal skin complications interfere with stoma appliance use and negatively affect patient quality of life. To find an alternative to long-term peristomal skin treatment involving corticosteroid products, a prospective study was conducted to compare the effect of a German chamomile solution to topical steroids on peristomal skin lesions in colostomy patients. Persons seeking care for the treatment of a peristomal skin lesion were assigned to a treatment regimen of once-a-day hydrocortisone 1% ointment (n = 36) or twice-a-day chamomile compress (n = 36) application. Treatments were assigned by matching patient demographic, history, and skin condition variables. At baseline, no significant differences between the variables were observed. Forty-two (42) of the 72 patients were female. Most participants had their stoma for more than 1 year (18.14 months in the chamomile and 17.69 months in the steroid group). Lesions were assessed every 3 days for a maximum of 28 days. Lesions healed significantly faster in the chamomile than in the hydrocortisone group (mean time to healing 8.89 +/- 4.89 and 14.53 +/- 7.6 days, respectively; P = 0.001). Stoma patient symptoms (pain and itching) also resolved more expediently in the chamomile than in the hydrocortisone group. Because corticosteroids are nonspecific anti-inflammatory agents, herbal extract use can prevent the side effects of long-term topical corticosteroid use. The results of this study suggest that German chamomile can be recommended to relieve itching and inflammation and that twice-daily application facilitates healing of peristomal skin lesions. Methods to facilitate the application of topical treatments without interfering with appliance adhesion or necessitating frequent appliance removal should be refined. Additional randomized studies are needed to confirm the results of this study. PMID- 21617263 TI - Adenocarcinoma of a colostomy following abdominoperineal resection for squamous cell carcinoma of the anal canal: a case study. AB - Malignant neoplasms presenting on a stoma, as well as the development of colorectal adenocarcinoma after previous treatment for squamous cell carcinoma (SCC) of the anal canal, are rare. The unique case is presented of an 81-year-old woman with parastomal bleeding and ulceration found to have a primary colorectal adenocarcinoma arising de novo on a colostomy, formed after salvage abdominoperineal resection (APR) 3 years earlier for recurrent anal SCC. This is the first reported case of a colonic adenocarcinoma on a colostomy formed after an APR for anal SCC. Although stomal neoplasia is rare, the appearance of a friable bleeding lesion on the stoma should be investigated to exclude metastatic cancer or a second primary malignancy. PMID- 21617265 TI - Management options for rare diseases in children and adolescents in Georgia (experience of the country with transitional economy). AB - We present the results of our research and organizational work aimed at the management optimization for the rare diseases in Georgia (the Country with Transitional Economy). We compiled a list of the rare diseases actual for Georgia; elaborated algorythms and expert systems supporting the diagnosis making process for various clusters of the rare diseases; translated into Georgian and adapted textual materials regarding the management of various rare disorders; assessed the awareness level for the rare diseases of the pediatricians and general practioners in Georgia and attempted to raise it by organizing seminars and conferences, including international ones, in various regions of Georgia; elaborated a model of the expert system (based on the fuzzy logic principles) for unmasking the cases suspicious for the rare disease; laid the foundation for the national register of the rare diseases in children and adolescents; elaborated the module for post-graduate education regarding rare diseases; organized the center for the rare diseases. PMID- 21617266 TI - Public health research on rare diseases. AB - Despite the low prevalence of Rare Diseases (RD), over 30 million EU citizens suffer from these conditions. This paper summarizes some aspects of these life threatening chronic and debilitating diseases that usually require long term specialist care and costly formal and informal surveillance. Epidemiology does have an important role to play in the field of RD, since it provides appropriate methods and tools for assessing exposures and health outcomes. In this regard, the utility of registries, biobanks and population-based surveillance systems are discussed. The lack of effective diagnoses and treatments in RD patients often underlies their shortened life expectancy and quality of life. Due to the limited number of patients and the scarcity of relevant knowledge and expertise, coordination at European level is probably the best way of pooling the very limited resources available and provides a very high added-value. RD require the combined efforts of health and social care professionals, politicians, managers and researchers to increase the availability of effective disease management tools to improve care and to extend both life expectancy and Health Related Quality of Life. PMID- 21617267 TI - General knowledge and awareness on rare diseases among general practitioners in Bulgaria. AB - Rare diseases are a serious public health problem and are a threat to the health of EU citizens. Important role in the area of rare diseases have the medical specialists who diagnose and monitor the course of the disease of each patient. General practitioners (GPs) are usually the first to identify "unusual" patients that might have a rare disease. The GPs awareness and knowledge about rare diseases is a strong factor for the timely and accurate diagnosis and adequate treatment of rare diseases conditions. A telephone interview was conducted among the GPs in Bulgaria between January and March 2008. A set of 10 questions with pre-defined answers was constructed and offered to the GPs in order to determine their level of knowledge and awareness of rare diseases. Data were statistically processed using specialized software SPSS version 9.0 (SPSS, Inc., Chicago IL). The responses of surveyed doctor indicate a low level of general knowledge and awareness. This means, that GPs from the primary health care system in Bulgaria could not provide sufficient in quality and timeliness specific information to their patients with rare diseases. A campaign for increasing the awareness of GPs about rare diseases is needed. PMID- 21617268 TI - [University center on rare diseases, as a successful example of the integration of the clinical sector into the system of medical education]. AB - There is substantiated appropriateness of the establishment of the University Center on Rare Diseases. It is indicated that the center could illustrate a successful integration of the clinical sector into the system of medical education. The model of the functioning of center and its regulations are presented. The main topics of the special teaching module on rare diseases for physicians are submitted as well. PMID- 21617269 TI - [Purposefulness of using fuzzy logic approaches in the rare disease clinical trials]. AB - Different methods being used in the rare diseases clinical trials are examined. There is shown the purposefulness of using fuzzy approaches for such studies. Some advantages of Fuzzy logic methods in comparison with Baysian approach are substantiated. PMID- 21617270 TI - Unilateral asymptomatic testis enlargement in children and adolescents. AB - In literature a well codified definition of unilateral asymptomatic testis enlargement does not exist. Therefore in these cases the pediatrician or adolescentologist will have to make a clinical and diagnostic evaluation in order to exclude: a) an enlarged testis secondary to tumors, surgery, or endocrinological diseases; b) a small testis due to a previous (ex. cryptorchidism) or current disease (e.g. varicocele).The presence of a mild difference in testis volumes during puberty is not at all rare. This situation may be due to the technique used for evaluation of testis volume or secondary to a varicocele. The identification of variants of testis enlargement is important, because, while on one hand there are conditions without clinical relevance, on the other hand, there are diseases that require early diagnosis and immediate treatment. The Authors report a brief review of the literature and their own clinical experience. 14 patients with unilateral testis enlargement were observed. At the first examination, mean age was 12.3+/-1.2 years and the volume of the enlarged testis varied between 4 ml and 20 ml (mean volume 10+/-4 ml) versus 1.5 ml and 10 ml (mean volume 5+/-2 ml) of the contralateral testis. In 75% of cases the right testis was affected. During the ten year follow-up, the volume of the enlarged testis never exceeded 25 ml and progressive reduction of the difference between the two testes was demonstrated. Therefore, they propose another clinical condition defined as transitory unilateral testis enlargement of puberty. PMID- 21617271 TI - [Clinical value of changes in red blood cell ultrastructure and energy metabolism in children with cystic fibrosis]. AB - In 42 patients of various age (from 1 month to 14 years) with cystic fibrosis were analyzed ultra structure, level of adenylnucleotides and activity ATP-ase of erythrocytes, in order to characterize their membrane and energy metabolism. The studies revealed the changes in erythrocytes in the cases of cystic fibrosis. In the cases of broncho pulmonary form of cystic fibrosis were detected I and II row echinocytes, cone-shaped erythrocytes, also erythrocytes with reach-through hole in center. At mixed form of cystic fibrosis were detected more changes in erythrocytes than in other forms of this disease. Both cone-shaped erythrocytes were more than in other forms of cystic fibrosis. Also there were detected erythrocytes with holes (round, polygonal) in their center. The results of the study provide a more precise diagnosis, in time correction of disorders and a comprehensive assessment of multiple-modality treatment of cystic fibrosis. PMID- 21617272 TI - [Oral cavity features in patients suffering from osteogenesis imperfecta]. AB - Osteogenesis Imperfecta (OI) is a rare hereditary connective tissue disorder. This pathology is characterized by disruption of biosynthesis of Type I collagen, and production of limited amount of defective and imperfect collagens. This causes decrease in bone mass of human body, bones become fragile and brittle, resulting in unreasonable multiple fractures. Reportedly, number of patients with OI ranges between 32-38 in Georgia. However, exact number of patients, including children and their parents, is unknown. Dentinogenesis Imperfecta (DI; DGI) and skeletal malocclusion occupy special place in varied spectrum of OI clinical symptoms. We studied 14 patients: 9 women (64.3%), 5 men (35.7%) and divided them in three age groups: I - 2.5-6 years - period of primary dentition (28.6%), II - 6-14 years - period of changing teeth dentition (35.7%) and III - above 14 years period of permanent dentition (35.7%). 28.5% of screened patients had one of the symptoms of DI, such as tooth discoloration. Discoloration of primary teeth was revealed in 4 patients (primary dentition). Another symptom of DI, such as early abrasion, was detected in 5 patients i.e. 35.71%. This was divided in the following manner: I age group - 3 cases, II and III age groups - 1-1 cases. It was also observed that early abrasion of primary teeth prevails over permanent. One of DI's radiographic symptoms, such as peculiar form of teeth crown and root, was revealed in 21.4% or in 3 patients, 2 of whom had bulbous crown, and the third one deformed (curved) root. Peculiar characteristics of DI, such as increased constriction of the coronal-radicular junction, obliterated pulp chamber, short and narrow roots, were not observed in the patients examined. Interesting characteristic of DI, such as periapical destruction of intact tooth root, was revealed in the form of bone defect in 7.1% of those examined (1 patient). Therefore, out of examined 14 patients with OI - DI had 6 patients or 42.85% of cases. Also, interesting observation was revealed - DI is more common in primary teeth (66.66%) than in permanent (33.33%). Radiographic examination - orthopantomography - revealed secondary osteoporosis of jaw bones in 100% of cases. Mucous tissue of examined patients is within normal range. Among examined patients, 1 case of adenty, 1 case of retention and 1 case of overcomplex tooth were revealed. According to current literature, it is unknown whether there is a lgical relationship between adenty, retention, overcomplex teeth and OI. This will be defined by future research. PMID- 21617273 TI - The Marfan syndrome - features, natural history and treatment options - our experiences. AB - The Marfan syndrome (MFS) is one of the most common (1:3000-1:4000) heritable connective tissue disorders. It's still a rarely diagnosed syndrome, especially in childhood. Near all cases MFS results from mutations in the fibrillin-1 (FBN1) gene on chromosome 15q21.1, which encodes for the glycoprotein fibrillin. The FBN1 gene is a large protein that can cause more than 500 mutations and molecular examinations, finally confirming the diagnosis, are conducted extremely rare. We present prospective data concerning 66 patients with clinically-diagnosed MFS who have been controlled in Department of Pediatric Cardiology and Congenital Heart Diseases Medical University in Gdansk in 2000 - 2010. 29 patients (44%) had mitral valve regurgitations, 19 (29%) aneurysmal dilatation of the aorta, 13 (20%) had both these irregularities. In 7 cases (11%) diagnosis of mitral valve prolapse preceded appearance of an aneurysmal dilalation of the aortic bulb. During the observation 11 patients (17%) underwent cardiosurgical procedures for the sake of stopping crucial progressive mitral valve dysfunction and/or aneurysmal dilatation of the aortic bulb, which threatened with a rupture of aortic aneurysm. In 39 cases (59%) prophylactic treatment with beta - blockers was administered. The patients with MFS need a multidisciplinary system of care and the psychological supporting. The cardiosurgical treatment, which nowadays is bringing better results, due to the technological advancements is a new hope for this patient population. PMID- 21617274 TI - Influence of halothane narcosis on a change of the number of GABA-positive cells in the hippocampus of adult rats with the midasolam premedication. AB - The most difficult questions in pediatric anesthesiology still remain relevant at surgeries to correct congenital maxilla-facial pathologies, accompanied by severe respiratory distress, speech and auditory sensation. For a long time the anesthetics were widely used among the neonatal and young children with extreme care due to their indicated suppressive effects on the immature organ systems. According to this fact, the choice of anesthesia is a matter of extremely importance. In addition, the study of molecular mechanisms indicating to the adverse effects induced by application of anesthesia will make it possible to develop therapeutic strategies for prevention of postoperative complications. The purpose of present study is to investigate a change of the quantity of GABA positive cells in the hippocampus of white adult rats under the conditions of halothane narcosis and with the premedication by midazolam. Subjects of a study - 30 adult white rats with the weight of 130-140 g. Animals were subdivided into three groups. I group - intact animals; II group - sham-operated rats under the conditions of halothane narcosis; III group - sham-operated animals, premedicated by midazolam, under the halothane narcosis. A change in the quantity of GABA positive cells was determined by the immunohistochemical method (Rabbit Anti GABA). All data were processed by the method of standard variation statistics. The authenticity of results was evaluated according to the Student's criterion within limits of 95-99% reliability. The results of conducted by us studies revealed, that during the application of halothane narcosis causes an increase in the GABA positive cells in the fields CA1 on hippocampus of adult white rats. In 24 hours after sham operation, at the condition of application of halothane narcosis a quantity of GABA positive cells raises in the CA3 field of hippocampus in II group animals as well. Moreover, in the CA1 field correlation between GAD65/67 positive and GABA positive cells is revealed which was not observed at later stages. The study of this question is the purpose of our next investigations. PMID- 21617275 TI - Endocrine histology findings in a prepubertal thalassemic girl with multiple endocrine complications secondary to iron overload. AB - beta-thalassaemia major (TM) is an inherited disorder of erythropoiesis requiring regular blood transfusions and chelation therapy for the iron overload resulting from transfusions and increased gastrointestinal absorption. Endocrine dysfunctions are common in older children with TM and has been attributed to iron deposition in endocrine glands. The Authors report the clinical and histological findings of endocrine glands in a prepubertal girl with multiple endocrine complications secondary to iron overloadn died from cardiac failure. Variations in severity of the disease and therapeutic regimens may result in different incidence and types of complications It is emphasized the importance of chelating therapy to protect endocrine glands from haemosiderosis. PMID- 21617276 TI - A novel hepatocyte nuclear factor-1beta (MODY 5) gene mutation in a Romanian boy with pancreatic calcifications, renal and hepatic dysfunction. AB - We report a 12-years-old Romanian boy with a diagnosis of diabetes and renal insufficiency. Mutations in homeodomain-containing transcription factor hepatocyte nuclear factor (HNF-1beta) have been reported in association with maturity-onset diabetes of the young (MODY 5) and early maturity-onset diabetes, progressive non-diabetic renal dysfunction and bilateral renal cysts. We found a new heterozygous mutation in HFN-1beta located in the exon 3 (c.715 G>C; p.239R) associated to pancreatic calcifications. The importance of molecular diagnosis of MODY patients is reinforced and the need for a careful follow-up is stressed in order to monitor the progression of clinical manifestations and its correlation with the gene mutation. PMID- 21617277 TI - The course of the dilated cardiomyopathy in three siblings - a rare case of familial non-compact left ventricle. AB - The dilated cardiomyopathy still remains a big problem in infant's cardiology. Almost a third of patients with this diagnosis die in infancy, 30% will suffer from the chronic heart failure that forces constant treatment or/and heart transplantations, and in remaining 30% we notice improvement during infancy. We presented the clinical course and progress of dilated cardiomyopathy based on the case of three siblings. Signs of the heart failure were nonspecific. Concluding: diagnostic vigilance must be shown in the cases of positive familial history. PMID- 21617278 TI - Hyperparathyroidism due to auto-immunological malabsorption in an African girl. AB - Hyperparathyroidism is a rare finding in children. It is a typical sign of vitamin D-deficiency caused by different reasons. It may also be due to calcium wasting syndromes, and it can rarely be induced by adenomas of the parathyroid glands and in parathormone receptor mutations (pseudohyperparathyroidism). A 12 year old Gambian girl living in Hamburg, Germany, was developing abdominal and joint pain. Serum analysis revealed low serum-calcium, significantly elevated parathormone and decreased vitamin D. Immigrant rickets was assumed. Because of abdominal pain and iron deficiency, lambliasis was ruled out. Celiac disease was demonstrated by gliadin and endomysium antibodies as well as by intestinal mucosa biopsy. Despite of a gluten-free diet the joint pains persisted. They were declared by rheumatologists to be caused by a chronic juvenile arthritis (sister disease of celiac disease). However, there were no positive inflammation signals and no clear elevated rheuma-immunology. Follow up: Gluten-free diet and additional treatment with calcium and active vitamin D did not stop increasing parathormone levels, did not stop abdominal and joint pain, and did not stop increment of positive celiac disease antibodies. Assuming compliance problems the patient was then treated with vitamin D injections, which caused decreasing parathormone levels and vanishing joint pain. Celiac disease can cause intestinal rickets with elevated parathomone levels mimicking chronic juvenile arthritis, if gluten-free diet is not strictly performed by compliance problems. PMID- 21617279 TI - Somatostatin - therapeutic option for chylothorax in preterm neonates. Report on two patients and review of the literature. AB - We report the use of somatostatin in two preterm neonates with chylothorax and discuss those cases in the light of current literature on the treatment of chylothorax in infants born preterm. Chylothorax, a severe complication in thorax surgery, is also a symptom of different diseases and may even occur spontaneously. Treatment is difficult, especially in preterm neonates with co morbidities. The standard therapeutic strategy with non-invasive procedures (e.g. enteral diet free of long chain triglycerides or parenteral nutrition) is not always effective. Surgical interventions, like pleurodesis, ligation of the ductus thoracicus, or pleuroperitoneal shunt may be of considerable risk in preterm infants and must be carefully evaluated. Somatostatin is a new non invasive therapeutic option for the treatment of chylothorax in adults and older pediatric patients. Case reports demonstrate the effectiveness of the somatostatin treatment, mostly in adult patients and in adolescents. There are only few case reports describing the use of somatostatin in preterm neonates. One VLBW (very low birth weight) and one hypotrophic ELBW (extremely low birth weight) neonate (gestational ages of 30+3/7, and 25+2/7 weeks; birth weights of 1270g, and 450g respectively) were treated for chylothorax with continuous infusion of somatostatin in addition to the dietary treatment. The chylothorax disappeared after start of somatostatin. No major side-effects of the somatostatin treatment were observed. As reported in other published pediatric cases, somatostatin seems to be a therapeutic option for the treatment of chylothorax in preterm neonates. In review of the literature we identified another eleven case reports on the treatment of persistent chylothorax with somatostatin or octreotide in preterm neonates. Further observations are needed before somatostatin can be recommended as a standard first-line treatment procedure for chylothorax in infants. PMID- 21617280 TI - Hallermann-Streiff syndrome: a case report from Georgia. AB - We report a 9 years and 6 month old boy with the Hallermann-Streiff syndrome (HSS). The patient was referred by a pediatrician. The diagnosis was established by endocrinologist based on the presence of specific facial gestalt (bird-like face) and bilateral congenital cataracts. The patient was not short, but had mild mental retardation; hypotrichosis was represented by Sign of Hertoghe (Queen Anne's sign) only. Thyroid function was normal. X-ray study yielded valuable data. The night apnoea (secondary to the dyscephalic narrowing of the upper airways) constituted the major concern. We also discuss diagnostic criteria for the HSS along with significance of various clinical signs. Combination of specific facial gestalt and ocular abnormalities should be particularly alarming. Endocrine aspects of the HSS are reviewed. X-ray study is recommended as an inexpensive and readily available but informative tool. PMID- 21617281 TI - [The congenital parcial erithroblastopenia--Diamond-Blackfan anemia]. AB - The article presents twenty-five year observation on 5 patients with Diamond Blackfan anemia. In 2010 the new case of this pathology in neonate was diagnosed. Research suggests that rarely, the Diamond-Blackfan anemia may be may be the result of an aplastic anemia. To find out what the real cause of anemia is the number of reticulocytes and qualitative and quantitative indicators bone marrow should be investigated in newborns and infants. PMID- 21617282 TI - [Thrombocytopenia-absent radius - TAR-syndrome]. AB - Thrombocytopenia-absent radius - TAR-syndrome is a rare condition in which thrombocytopenia is associated with bilateral radial aplasia. TAR syndrome was first described in 1956 by H. Cross et al. An autosomal recessive inheritance pattern was proposed because TAR affected more than one member of some families. The disease varies and includes abnormalities in the skeletal, hematological, cardiac, gastrointestinal and other systems. In article the 30 day infant with thrombocytopenia-absent radius - TAR-syndrome is described. The diagnose was confirmed by clinical, laboratory, radiological, genetic etc. investigations. PMID- 21617283 TI - A case of DiGeorge syndrome in Georgia. AB - Patient 6 - year- old boy, with history of recurrent otitis, cleft palate, was admitted to the hospital for fever, abdominal pain; He had high ESR,CRP, low T lymphocytes, VSD. Peritoneal fluid was positive for pseudomona aeroginoza. Diagnosis of DiGeorge syndrome was confirmed by further genetical study. Immune deficiencies should be considered when infections are severe, persistent resistant to standard treatment, or caused by opportunistic organisms. Treatments can often correct many of the critical and immediate problems associated with DiGeorge syndrome such as heart defects, calcium defects, poor immune system functions and cleft palate. People who had poor immune function as children due to small or missing thymus, may have an increased risk of autoimmune disorders, such as a rheumatoid arthritis and Graves disease. Because DiGeorge syndrome can result in so many disorders, a number of specialists should be involved in diagnosing specific conditions, recommending treatments and providing care. PMID- 21617284 TI - Pediatric drug induced hypersensitivity syndrome: case report. AB - Drug induced hypersensitivity syndrome (DIHS) is a rare but potentially life threatening adverse drug reaction particularly associated with anticonvulsant drugs. We describe the case of a patient who suffered from DIHS caused by anticonvulsant. Our experience suggests glucocorticoids (GC) may be of benefit to children with severe DIHS. PMID- 21617285 TI - Rare tubulopathy - primary hypophosphatemic rickets (case report). AB - Primary hypophosphatemic rickets is a rare disorder caused by inborn defect of renal tubular reabsorbtion and usually manifested in childhood and infancy with stunted growth and deformities of lower limbs. Patient 12 years old, female, was born to healthy parents, at 41 weeks of gestation by normal delivery and had a normal birth weight and length. She had one healthy sibling (male). Psychomotor development until the age of 2 years was normal. Since the age of 2 years deformation of legs and difficulties with walking have been observed. The Patient was consulted by Pediatric Orthopedist, Nephrologists and Endocrinologist. The blood biochemical findings revealed normal pH, normal calcium (CA), potassium (K), sodium (Na) concentrations, very low phosphate (P) with markedly elevated alkaline phosphatase (AP) and slightly elevated parathyroid hormone (PTH) concentration; urine test indicated impaired tubular function: mild glucosuria, proteinuria, and markedly increased phosphaturia. Phosphate tubular reabsorbtion (PTR) was 52%, phosphate/creatinine clearance 0,52, renal threshold phosphate concentration (TmP/GFR) 0,25 mmol/l. Genetic tests results not available yet. RESULTS: the diagnosis of phosphate diabetes made on the basis of clinical laboratory data. The therapy with Inorganic phosphate (50-100mg/kg/d) and 1,25(OH)D3 40-50 ng/kg/d had been started. Childs walking abilities improved with treatment, the deformation of legs decreased, but O-legs and other skeletal deformations are still remarkable. Auxological parameters are not satisfactory. Physical growth is below the normal range (height SDS -3,78); the patient's predicted height less than target height. Biochemical monitoring performed regularly under the treatment, serum phosphate remains below normal, serum ALP is still elevated. CONCLUSION: This case is in line with other publications and indicates the difficulty to achieve normal phosphate levels and normal growth without an additional treatment with growth hormone. At this stage of the disease, the question of referring to the growth hormone therapy is being considered aimed to improve the growth tendencies. It is possible that orthopedic surgery is also needed in the future. PMID- 21617286 TI - Encephalocele and skeletal malformations (case report). AB - Encephalocele is a herniation of the brain (cranium bifidum, cephalocele, craniocele), formed during embryonic development, because of the incomplete closure of Neural Tube. It is a rare skull defect, with the incidence of 0.8 to 5 per 10,000 live births. The article presents the medical history of a four month old patient, with frontoethmoidal encephalocele and multiple skeletal anomalies, such as amniotic knots on limbs, foot deformity, sindactilia and cleft palate. PMID- 21617287 TI - International Conference on Rare Diseases in Tbilisi, 2010. PMID- 21617289 TI - A model for filtered backprojection reconstruction artifacts due to time-varying attenuation values in perfusion C-arm CT. AB - Filtered backprojection is the basis for many CT reconstruction tasks. It assumes constant attenuation values of the object during the acquisition of the projection data. Reconstruction artifacts can arise if this assumption is violated. For example, contrast flow in perfusion imaging with C-arm CT systems, which have acquisition times of several seconds per C-arm rotation, can cause this violation. In this paper, we derived and validated a novel spatio-temporal model to describe these kinds of artifacts. The model separates the temporal dynamics due to contrast flow from the scan and reconstruction parameters. We introduced derivative-weighted point spread functions to describe the spatial spread of the artifacts. The model allows prediction of reconstruction artifacts for given temporal dynamics of the attenuation values. Furthermore, it can be used to systematically investigate the influence of different reconstruction parameters on the artifacts. We have shown that with optimized redundancy weighting function parameters the spatial spread of the artifacts around a typical arterial vessel can be reduced by about 70%. Finally, an inversion of our model could be used as the basis for novel dynamic reconstruction algorithms that further minimize these artifacts. PMID- 21617290 TI - A robust procedure for verifying TomoTherapy Hi-ArtTM source models for small fields. AB - The dosimetric measurement and modeling of small radiation treatment fields (<2 * 2 cm2) are difficult to perform and prone to error. Measurements of small fields are often adversely influenced by the properties of the detectors used to make them. The dosimetric properties of small fields have been difficult to accurately model due to the effects of source occlusion caused by the collimating jaws. In this study, small longitudinal slice widths (SWs) of the TomoTherapy(r) Hi-Art(r) machine are characterized by performing dosimetric measurements topographically. By using a static gantry, opening the central 16 MLC leaves during the irradiations, and symmetrically scanning detectors 10 cm through each longitudinal SW, integral doses to a 'TomoTherapy equivalent' 10 * 10 cm2 area are topographically measured. To quantify the effects of source occlusion for TomoTherapy, a quantity referred to as the integral scanned dose to slice width ratio (D/SW) is introduced. (D/SW) ratios are measured for SWs ranging from 0.375 to 5 cm in size using ion chambers and a radiographic film. The measurements of the (D/SW) ratio are shown to be insensitive to the detectors used in this study. The (D/SW) ratios for TomoTherapy have values of unity in the range of SW sizes from 5 cm to approximately 2 cm. For SWs smaller than 2 cm in size, the source occlusion effect substantially reduces the measured machine output and the value of the (D/SW) ratios. The topographic measurement method presented provides a way to directly evaluate the accuracy of the small-field source model parameters used in dose calculation algorithms. As an example, the electron source spot size of a Penelope Monte Carlo (MC) model of TomoTherapy was varied to match computed and measured (D/SW) ratios. It was shown that the MC results for small SW sizes were sensitive to that particular parameter. PMID- 21617291 TI - Small animal radiotherapy research platforms. AB - Advances in conformal radiation therapy and advancements in pre-clinical radiotherapy research have recently stimulated the development of precise micro irradiators for small animals such as mice and rats. These devices are often kilovolt x-ray radiation sources combined with high-resolution CT imaging equipment for image guidance, as the latter allows precise and accurate beam positioning. This is similar to modern human radiotherapy practice. These devices are considered a major step forward compared to the current standard of animal experimentation in cancer radiobiology research. The availability of this novel equipment enables a wide variety of pre-clinical experiments on the synergy of radiation with other therapies, complex radiation schemes, sub-target boost studies, hypofractionated radiotherapy, contrast-enhanced radiotherapy and studies of relative biological effectiveness, to name just a few examples. In this review we discuss the required irradiation and imaging capabilities of small animal radiation research platforms. We describe the need for improved small animal radiotherapy research and highlight pioneering efforts, some of which led recently to commercially available prototypes. From this, it will be clear that much further development is still needed, on both the irradiation side and imaging side. We discuss at length the need for improved treatment planning tools for small animal platforms, and the current lack of a standard therein. Finally, we mention some recent experimental work using the early animal radiation research platforms, and the potential they offer for advancing radiobiology research. PMID- 21617292 TI - Radon potential mapping of the Tralee-Castleisland and Cavan areas (Ireland) based on airborne gamma-ray spectrometry and geology. AB - The probability of homes in Ireland having high indoor radon concentrations is estimated on the basis of known in-house radon measurements averaged over 10 km * 10 km grid squares. The scope for using airborne gamma-ray spectrometer data for the Tralee-Castleisland area of county Kerry and county Cavan to predict the radon potential (RP) in two distinct areas of Ireland is evaluated in this study. Airborne data are compared statistically with in-house radon measurements in conjunction with geological and ground permeability data to establish linear regression models and produce radon potential maps. The best agreement between the percentage of dwellings exceeding the reference level (RL) for radon concentrations in Ireland (% > RL), estimated from indoor radon data, and modelled RP in the Tralee-Castleisland area is produced using models based on airborne gamma-ray spectrometry equivalent uranium (eU) and ground permeability data. Good agreement was obtained between the % > RL from indoor radon data and RP estimated from eU data in the Cavan area using terrain specific models. In both areas, RP maps derived from eU data are spatially more detailed than the published 10 km grid map. The results show the potential for using airborne radiometric data for producing RP maps. PMID- 21617293 TI - Radiological indices of technologically enhanced naturally occurring radionuclides: a PIXE approach. AB - This paper reports an assessment of the level of the radionuclides (40)K, (232)Th and (238)U in environmental soil samples (process waste), and hence their calculated dose rates. For this purpose, the radioactivity from three natural radionuclides was determined in tin process-waste samples in Jos, Nigeria. This work is based on the particle induced x-ray emission (PIXE) approach, devoid of the secular equilibrium, and most of the resolution, interference, self absorption, geometrical and density correction problems inherent in gamma spectrometry. Many potential environmental hazards have been observed and the data would be of use to the government in its remediation plan for the study area. The high hazard indices require taking adequate measures to check exposures, and an underground lining in the waste ponds to prevent direct contact with the waste pile is recommended. The use of the wastes as building materials should be stopped and use of soils around this area in any development projects should be discouraged until detailed studies on indoor radiation doses and the effects on the inhabitants of prolonged exposures have been carried out. PMID- 21617296 TI - Severe deterministic effects of external exposure and intake of radioactive material: basis for emergency response criteria. AB - Lessons learned from responses to past events have shown that more guidance is needed for the response to radiation emergencies (in this context, a 'radiation emergency' means the same as a 'nuclear or radiological emergency') which could lead to severe deterministic effects. The International Atomic Energy Agency (IAEA) requirements for preparedness and response for a radiation emergency, inter alia, require that arrangements shall be made to prevent, to a practicable extent, severe deterministic effects and to provide the appropriate specialised treatment for these effects. These requirements apply to all exposure pathways, both internal and external, and all reasonable scenarios, to include those resulting from malicious acts (e.g. dirty bombs). This paper briefly describes the approach used to develop the basis for emergency response criteria for protective actions to prevent severe deterministic effects in the case of external exposure and intake of radioactive material. PMID- 21617297 TI - Effective dose: practice, purpose and pitfalls for nuclear medicine. AB - Effective dose (E) is the only comparatively simple dose quantity that is related to health detriment for stochastic effects from exposure to ionising radiation. As such, E has found wide application for medical exposures, as it allows comparisons with doses from different examinations and other sources. E is derived from the weighted sum of doses to tissues known to be sensitive to radiation from epidemiological studies and contains inherent approximations. Thus it is not a scientific quantity, but a practical one that the International Commission on Radiological Protection (ICRP) has created for use in the calculation of reference doses for protection purposes. In the application of E to medical exposures, there has been a tendency to attribute a greater accuracy to values of E than is justified by its derivation. Recognising that E is strictly not subject to uncertainties, an analysis has been undertaken of potential uncertainties in E for different nuclear medicine examinations to enable users to judge its reliability as a comparator of relative risk. Assessments have been based on the considered accuracy of the component parts and indicate that the uncertainties in the values of E as a relative indicator of harm for nuclear medicine procedures for a reference patient are about +/- 50%. These are larger than those for radiology procedures, because of the tendency for doses to single organs, especially the bladder, to form a substantial part of E for some procedures. Revision of the tissue weighting factors in 2007 produced a 10% decrease in the mean value of E for nuclear medicine examinations. Estimations of cancer risk based on E for an individual could vary by one or two orders of magnitude. E fulfils an important role as a health-related dose quantity that can be used in justification of nuclear medicine examinations, but physicians should be aware of its limitations. General terminology should be used in conveying risks to patients and medical professionals. PMID- 21617298 TI - The history of IRPA--up to the millennium. AB - Since its foundation, the International Radiation Protection Association (IRPA) has grown to become the international voice of the profession of radiological protection. When it came into being in 1965 the existence of such a profession had only recently been recognised, but already there was seen to be a need for coordination and exchange of information and views internationally. As with all such organisations, the early years and decades were taken up with the development and evolution of the organisation, but there comes a time when, if the history is not documented, it gradually gets lost. Recognising this, the Executive Council (EC) created the role of archivist and historian, and appointed Geoff Webb with a brief to compile the history of IRPA. This article is the result and covers the developments leading up to the formation of IRPA and from then to the year 2000. An expanded version with more names and photographs is available through IRPA. PMID- 21617299 TI - Effectiveness and safety of pandemic influenza A (H1N1) 2009 vaccine in healthcare workers at a university hospital in Japan. AB - Pandemic influenza A (H1N1) virus (AH1pdm) emerged in April 2009. An inactivated, split-virus, unadjuvanted AH1pdm vaccine was manufactured in Japan, and vaccination was initiated with top priority for healthcare workers (HCWs) on October 19, 2009. A retrospective cohort study was conducted to evaluate the effectiveness of a single-dose vaccine for HCWs in a hospital in Japan. A total of 1,567 (84.5%) of 1,854 HCWs were vaccinated. Thirty-seven were infected with AH1pdm before the vaccine became available, and were excluded. The other 250 were not vaccinated for personal reasons. We analyzed the influenza infection rate with or without vaccination and related adverse events. Among the 1,817 HCWs without previous infection, 37 were infected with AH1pdm; 13 (5.2%) of 250 unvaccinated HCWs became infected, which was a significantly higher rate than the 24 (1.5%) of 1,567 vaccinated HCWs (P=0.001). Multivariate analysis revealed that age of 20-29 years was a risk factor for infection (adjusted odds ratio [aOR], 3.7; P<0.001), and that vaccination was a preventive factor (aOR, 0.20; P<0.001). Adverse events occurred in 549 of 1,060 HCWs, but most were mild. Although vaccination was carried out during AH1pdm epidemic expansion, the single-dose AH1pdm vaccine proved effective in HCWs, and severe adverse events were rare. PMID- 21617300 TI - The adverse events of influenza A (H1N1) vaccination and its risk factors in healthcare personnel in 18 military healthcare units in Korea. AB - In this study, we characterized adverse events related to influenza A (H1N1) vaccination and studied the factors that influence the occurrence of these events. A total of 4,302 personnel in 18 military healthcare units in Korea received 0.5 ml of inactivated H1N1 vaccine. The study questionnaires were answered by 3,939 (91.6%) personnel, at both 2 weeks and 4 weeks after vaccination. Among these subjects, 3,531 (82.1%) who responded to all questions in the questionnaire were studied. After immunization, military doctors were ordered to report the occurrence of any adverse event related to the vaccine for 2 months. According to the responses of the subjects, the most prevalent events were fatigue (11.3%), pain at the injection site (8.38%), and myalgia (6.97%). Female gender, being in the age range of 20-49 years, obesity, regular alcohol consumption, and comorbidity, but not smoking status or pregnancy, were related to a high incidence of local or systemic adverse events after H1N1 vaccination. A total of 14 cases of adverse events were reported by the military doctors. In most reported cases, the subjects had fever in addition to the primary adverse event, and one patient was diagnosed with pneumonia. In conclusion, the overall burden of adverse events related to influenza A (H1N1) vaccination was not inconsequential, but most symptoms were mild. Female gender, middle-age range of 20-49 years, obesity, regular alcohol consumption, and comorbidity were risk factors for the occurrence of adverse events after H1N1 vaccination. PMID- 21617301 TI - Immunogenicity and safety of a China-made monovalent pandemic (H1N1) 2009 influenza A vaccine in healthcare workers in Guangzhou, China. AB - Because healthcare workers played an important role in the battle against novel pandemic (H1N1) 2009 influenza, a clinical study was conducted to examine the immunogenicity and safety of a single dose of a China-made monovalent, split virus, pandemic (H1N1) 2009 influenza vaccine in this special high-risk population. Healthcare workers in the First Affiliated Hospital of Guangzhou Medical College who received the pandemic (H1N1) 2009 influenza vaccine were prospectively enrolled. Antibody titers were measured at enrollment and 14 days later using hemagglutination-inhibition (HI) and microneutralization assays. Adverse events were recorded within 7 days and 6 months after vaccination. Double sera were provided by 51 of 65 enrolled subjects. Postvaccination titers of 1:40 or more on HI assay were observed in 96% of recipients. Seroconversion or a significant increase in titer on HI assay occurred in 59% of subjects. The factor increase in GMTs was 4.3. The majority of complaints were mild to moderate in intensity. Although more than half of healthcare workers seemed immune to the pandemic (H1N1) influenza A virus before vaccination, most of the subjects still showed a fast, robust immune response to a single 15-MUg dose of a monovalent, split-virus unadjuvanted pandemic H1N1 2009 influenza vaccine. PMID- 21617303 TI - Detection of group a rotavirus RNA and antigens in serum and cerebrospinal fluid from two children with clinically mild encephalopathy with a reversible splenial lesion. AB - We report on two children with mild encephalopathy with a reversible splenial lesion associated with group A rotavirus (GARV) infection. We examined stool, serum, and cerebrospinal fluid samples to determine the presence of the GARV VP7 gene and GARV antigen by reverse-transcription PCR and enzyme-linked immunosorbent assay, respectively. GARV antigen was detected in stool samples from both patients. The GARV G genotype was G9 in one child and G3 in the other. GARV antigens were also found in both serum samples. However, the GARV VP7 gene was detected in only one serum sample, which was collected on the first day of symptomatic illness. Neither GARV antigen nor the VP7 gene was detected in cerebrospinal fluid samples. Both patients had excellent outcomes. Our results suggest that the reversible splenial changes in our patients might have been caused by indirect effects to the central nervous system subsequent to viral infection. PMID- 21617302 TI - Genetic Analysis and Phylogenetic Characterization of Pandemic (H1N1) 2009 influenza viruses that found in Nagasaki, Japan. AB - Isolation and determination of the nucleotide sequence of hemagglutinin (HA) of the pandemic (H1N1) 2009 influenza viruses found in Nagasaki, Japan, were conducted. The alignment results of the predicted HA amino acid sequences of these strains compared to the known global isolates revealed 5 specific amino acid differences located within the antigenic sites. The phylogenetic analyses revealed that the majority of the Nagasaki isolates could be classified into 6 phylogenetic clusters. Almost all isolates collected in the early season were classified into cluster I, which apparently originated from A/Nagasaki/HA-6/2009 isolated from a patient who returned from the Philippines. This cluster ceased to spread after November 2009. Between the end of August 2009 and January 2010, 5 new phylogenetic clusters (II-VI) emerged with viruses from different origins, and cluster III continuously advanced until March 2010. These results suggest that the onset of the influenza epidemic in Nagasaki originated from patient(s) who returned from the Philippines, and subsequently, various imported strains from different origins sustained the virus spread. Among the Nagasaki isolates, A/Nagasaki/HA-58/2009 having an H275Y mutation in the neuraminidase gene, which confers resistance to oseltamivir, was isolated. This is the first report in which an oseltamivir-resistant pandemic H275Y mutant was identified in Nagasaki Prefecture. PMID- 21617304 TI - Epidemiological investigation of measles in sera of healthy people in Heilongjiang Province, China. AB - Serological samples of healthy people were collected to obtain the levels of measles antibodies in different groups of people in Heilongjiang Province, China. Using quantitative enzyme-linked immunosorbent assays to measure the antibody levels, we found the lowest antibody positive rate and the lowest geometric mean concentration values in healthy people aged 15-39. This group is the population at high risk for adult measles in Heilongjiang Province, and is the focus of measles elimination work. The new challenges for the eventual elimination of measles will be to address immunization strategies in this segment of the population in order to control the incidence. PMID- 21617305 TI - Evaluation of a quantitative real-time PCR assay for the detection of JC polyomavirus DNA in cerebrospinal fluid without nucleic acid extraction. AB - The JC polyomavirus (JCV) is the causative agent of progressive multifocal leukoencephalopathy (PML), a fatal demyelinating disease. The current diagnostic standard for PML is real-time PCR testing of extracted DNA for assessing the presence of JCV DNA in cerebrospinal fluid (CSF). This study was aimed at evaluating the feasibility of a real-time PCR assay without nucleic acid extraction for the rapid quantification of JCV DNA in CSF. CSF samples were heat treated or treated with DNAzol Direct, a commercially available reagent for direct PCR, and the performances of the real-time PCR assays using templates obtained by either treatment were compared with that using DNA extracts. JCV DNA was detected in the heat- or DNAzol Direct-treated samples containing only a few copies of the viral genome per reaction, and a linear relationship was noted between the copy number detected and the amount of input virus ascertained by the DNA extraction method. The sensitivities of the assays using the heat and DNAzol Direct treatments were 85.7 and 90.5%, respectively, with the results of the DNA extraction method being used as reference. These data demonstrate that the real time PCR assay introduced in this study can serve as a rapid and cost-effective method of testing for JCV without DNA extraction and thereby facilitate the assessment of PML. PMID- 21617306 TI - First detection of a putative knockdown resistance gene in major mosquito vector, Aedes albopictus. AB - The Asian tiger mosquito, Aedes albopictus (Skuse), is the major vector of Chikungunya fever and the secondary vector of dengue fever. We collected Ae. albopictus from Singapore and performed genotyping assay to detect mutations of the voltage-gated sodium channel, which is the target site of pyrethroid insecticides. We detected an amino acid substitution, F1534C, which is suspected to confer knockdown resistance (kdr) to pyrethroid insecticides. Of the collected mosquitoes, 53.8% were homozygous for this mutation, and the allele frequency of this mutation was estimated to be 73.1%. No kdr mutation was detected in the 5 other loci of domains II and IV. This is the first evidence for the presence of the kdr gene in Ae. albopictus, and our findings highlight the need for studying the global distribution of this allele in this important vector insect. PMID- 21617307 TI - Molecular epidemiology and antimicrobial resistance determinants of multidrug resistant Acinetobacter baumannii in five proximal hospitals in Taiwan. AB - We investigated the molecular epidemiology, antibiotic susceptibility, and antimicrobial resistant gene determinants of 23 multidrug-resistant (MDR) Acinetobacter baumannii samples that were collected from 5 proximal hospitals in Taiwan during April and May 2009. Major antibiotic resistance varied from 82.6 to 100%. Fivw pulsotypes were observed to spread clonally among the 5 hospitals. PCR screening revealed high distributions of intI1 (91%), bla(OXA-23) (57%), bla(ampC) (100%), adeB (100%), adeJ (100%), and abeM (100%) genes, which were prevalent in the MDR A. baumannii isolates. Resistance gene expression was examined by reverse transcription-PCR, and showed that increased ampC expression was associated with ceftazidime resistance, but expression of adeB, adeJ, or abeM did not guarantee antimicrobial resistance phenotypes. In addition, imipenem resistance in some A. baumannii strains may be mainly modulated by genes other than bla(OXA-51-like). This is the first direct evidence indicating local spread of MDR A. baumannii in Taiwan. The resistance gene determinants are widely distributed in clonal and nonclonal-related isolates. PMID- 21617308 TI - SpeI restriction enzyme displays greater discriminatory power than XbaI enzyme does in a pulsed-field gel electrophoresis study on 146 clinical Burkholderia pseudomallei isolates. AB - Restriction enzymes SpeI and XbaI were used in a pulsed-field gel electrophoresis (PFGE) study for molecular characterization of 146 clinical Burkholderia pseudomallei isolates. The PFGE parameters were optimized to enable comparable, reproducible, and robust results. The optimized parameters for both SpeI and XbaI restriction enzymes used in this study were 200 V and a pulse time of 5 to 65 s for a 28-h runtime. Using SpeI, 9 different clusters were identified, whereas 6 clusters were identified by XbaI digestion, which exhibited 85% similarity to SpeI. SpeI (discrimination index [D]=0.854) showed higher discriminatory power than XbaI did (D=0.464). PMID- 21617309 TI - First report on isolation and molecular characterization of clinical Mycobacterium setense isolates in Asia. AB - We herein present the third set of documented clinical Mycobacterium setense cases. Three clinically unrelated isolates were identified and characterized using various key conventional and molecular diagnostic tests. Phenotypic and molecular data analysis, particularly 16S rDNA, hsp65, and rpoB sequencing, provided evidence of M. setense involvement in clinical infections in Iranian patients. Our findings may shed light on the capability of this rare Mycobacterium sp. to cause infection in both healthy and immunocompromised patients. PMID- 21617310 TI - Phylogenetic analysis of an off-seasonal influenza virus A (H3N2) in Niigata, Japan, 2010. AB - The objective of this study was to characterize the off-seasonal influenza virus A subtype H3N2, which caused an outbreak in an elderly hospital in Niigata, Japan. Virus isolates were subtyped by the hemagglutination-inhibition test and screened for antiviral drug sensitivity by real-time PCR using cycling probe technology the and 50% inhibitory concentration (IC(50)) method. Whole genome sequencing was performed in order to determine the phylogeny of the outbreak virus. Seven virus isolates were analyzed in this study, and the results showed that all belonged to the influenza virus A (H3N2). These viruses exhibited the S31N mutation in M2, which confers resistance to amantadine. The results of the IC(50) analysis showed that these viruses were sensitive to both oseltamivir and zanamivir. Whole genome analysis revealed that the virus was similar to the A/Perth/16/2009 strain and that it is a triple reassortant virus with a 3+3+2 pattern of segment recombination. PMID- 21617311 TI - Relationship between RANTES polymorphisms and respiratory syncytial virus bronchiolitis in a Japanese infant population. AB - Respiratory syncytial virus (RSV) is the most important virus associated with bronchiolitis in infants and young children. The regulated upon activation, normal T-cell expressed and secreted protein (RANTES, also known as CCL5) appears to be a key player in the etiology of RSV-infected airway inflammation. In this study, we genotyped three single-nucleotide polymorphisms in the RANTES gene: 403G/A, -28C/G, and In1.1T/C in 59 infants with severe RSV bronchiolitis and 201 control subjects. The frequencies of the -403G/A+A/A, -28C/G+G/G, and In1.1T/C+C/C genotypes were significantly lower in patients with severe RSV bronchiolitis than in control subjects, and the frequencies of the -403A, -28G, and In1.1C alleles were significantly lower in RSV patients than in control subjects. The present results suggest that RANTES polymorphisms may confer risk for severe RSV bronchiolitis. PMID- 21617312 TI - FTA card utility for PCR detection of Mycobacterium leprae. AB - The suitability of the FTA(r) elute card for the collection of slit skin smear (SSS) samples for PCR detection of Mycobacterium leprae was evaluated. A total of 192 SSS leprosy samples, of bacillary index (BI) 1 to 5, were collected from patients attending two skin clinics in Myanmar and preserved using both FTA(r) elute cards and 70% ethanol tubes. To compare the efficacy of PCR detection of DNA from each BI class, PCR was performed to amplify an M. leprae-specific repetitive element. Of the 192 samples, 116 FTA(r) elute card and 112 70% ethanol samples were PCR positive for M. leprae DNA. When correlated with BI, area under the curve (AUC) values of the respective receiver-operating characteristic curves were similar for the FTA(r) elute card and ethanol collection methods (AUC=0.6). Taken together, our results indicate that the FTA(r) elute card, which enables the collection, transport, and archiving of clinical samples, is an attractive alternative to ethanol preservation for the detection of M. leprae DNA. PMID- 21617313 TI - Prevalence of class A and AmpC beta-lactamases in clinical Escherichia coli isolates from Pakistan Institute of Medical Science, Islamabad, Pakistan. AB - In this study, 121 Escherichia coli samples isolated from clinical specimens obtained from Pakistan Institute of Medical Science, Islamabad, Pakistan, were analyzed for extended-spectrum beta-lactamases (ESBLs) and AmpC beta-lactamases using disk-diffusion assay and polymerase chain reaction. Of the isolates, 78 and 43 were identified as ESBL and AmpC producers, respectively. The highest resistance (89%) was observed against cefotaxime, followed by ciprofloxacin (87.6%) and cefepime (87%). Genetic analysis showed the presence of different class A and class C beta-lactamase genes, either alone (44.7%) or in combination (53.6%). CTX-M (57.7%) was the most prevalent among class A, followed by TEM (20.3%) and SHV (15.4%). CIT (including LAT-1 to LAT-4, CMY-2 to CMY-7, and BIL 1) and MOX (including MOX-1, MOX-2, CMY-1, and CMY-8 to CMY-11) family-specific plasmid-mediated AmpC beta-lactamases were the most prevalent among these isolates. Our study showed that both class A and class C beta-lactamases contributed to cephalosporin resistance in the E. coli isolates, thereby limiting therapeutic options. Co-expression of these enzymes may further hinder the identification of ESBLs, which is a critical step for designing a successful treatment for multidrug-resistant E. coli. PMID- 21617314 TI - Molecular characterization of rifampicin- and isoniazid-resistant Mycobacterium tuberculosis strains isolated in Kazakhstan. AB - Kazakhstan is one of the 14 countries with a high rate of morbidity due to multidrug-resistant tuberculosis (MDR TB) in WHO European region. The aim of our study was to characterize mutations associated with drug resistance to rifampicin and isoniazid in Mycobacterium tuberculosis isolates from Kazakhstan. M. tuberculosis strains were isolated from TB patients in different regions of Kazakhstan. A drug susceptibility test was performed on Lowenstein-Jensen medium using the absolute concentration method. Sequencing analysis was performed of the rpoB rifampicin resistance-determining region and the katG gene, the oxyR-ahpC intergenic region, and the inhA promoter region in 259 MDR M. tuberculosis isolates, in 51 isoniazid-resistant isolates, and in 13 rifampicin-resistant isolates. The mutational analysis revealed that the most frequent mutations associated with rifampicin and isoniazid resistance in M. tuberculosis are the substitutions at codons 531 (82.7%) and 315 (98.4%) in the rpoB and katG genes, respectively. In addition, we have found mutations with lower frequency at codon 526 (8.4%), 533 (1.5%), and 516 (1.1%) in the rpoB gene. In 6.2% of the isolates, no mutations were found in the rpoB gene. The findings of this study provide useful data for a better understanding of the mutation spectrum of isoniazid and rifampicin resistance among strains isolated from patients in Kazakhstan. Our results are also useful for the development of diagnostic tests of MDR M. tuberculosis. PMID- 21617315 TI - Characterization of splenic cells during the early phase of infection with neuropathogenic mouse hepatitis virus. AB - The highly neuropathogenic cl-2 and less virulent srr7 viruses isolated from the neurotropic JHM strain of the mouse hepatitis virus exhibit super acute spread of virus (SAS), a term applied when rapid viral spread from an organ or part of the initially infected site to another non-adjacent organ or part is detected within 12 h after infection. Herein, we used a cytospin procedure to confirm SAS in splenic cells derived from mice whose brains were infected with these viruses. The cytospin procedure enabled effective preservation of the cells on glass slides. With this method, we could characterize extremely low populations of infected cells in the spleen (less than 0.1%) at 12 h post-inoculation with srr7. We observed that all kinds of splenic cells examined were infected, including B220(+)Ly-6C(+) plasmacytoid dendritic cells. The population of viral antigen positive splenic cells was only slightly higher in cl-2 infection than in srr7 infection, but the cells showing viral production were present in numbers significantly higher in cl-2 infection compared with srr7 infection. PMID- 21617316 TI - Epidemiological investigation and analysis of hepatitis A virus genomes in the three cases of hepatitis A infections that occurred in April-May 2010. PMID- 21617317 TI - First detection of measles virus genotype g3 in a Japanese woman: an imported case. PMID- 21617318 TI - Contribution of acquired factors to the pathogenesis of dilated cardiomyopathy. The cause of dilated cardiomyopathy: genetic or acquired? (Acquired-Side)-. AB - Although genetic abnormalities play a pivotal role in the development of dilated cardiomyopathy (DCM), acquired infection and autoimmune abnormalities, or both, appear to be predominant underlying disorders. Of these, viral infection causes target organ damage via perforin produced by suppressor T cells. Thereafter, various antigens released from damaged myocytes are presented on the major histocompatibility complex II, which is expressed in antigen-presenting cells, resulting in activation of both cellular (Th1) and humoral (Th2) immunity. Various antimyocardial antibodies are detected in the serum of patients with DCM and recent findings suggest that at least some of them are directly related to the pathophysiology of DCM. An autoantibody targeting the beta1-adrenergic receptor is related to the persistent myocardial damage resulting in DCM and provides the substrate for fatal ventricular arrhythmias. An antibody for the muscarinic M2 receptor is related to atrial fibrillation, an antibody targeting Na-K-ATPase is closely related to sudden cardiac death from fatal ventricular arrhythmias, and an autoantibody for troponin I increases the L-type calcium current and is related to myocardial damage. On the other hand, genetic factors are also involved in susceptibility to viral infection and aberrations of acquired immunity, including antigen presentation and autoantibody production. In conclusion, acquired factors are predominant causes of DCM, although the 2 predisposing factors are also linked to genetic abnormalities. PMID- 21617319 TI - Contribution of genetic factors to the pathogenesis of dilated cardiomyopathy: the cause of dilated cardiomyopathy: genetic or acquired? (genetic-side). AB - Dilated cardiomyopathy (DCM) is characterized by dilated ventricles and systolic dysfunction. Its etiology is not fully unraveled, but both extrinsic and intrinsic factors are considered to be involved. The intrinsic factors include genetic variations in the genes (ie, disease-causing mutations and disease associated polymorphisms), which play key roles in controlling the susceptibility to the disease by affecting the performance, regulation, and/or maintenance of cardiac function. DCM can be classified into 2 types: hereditary and non hereditary. The genetic variations, or disease-causing mutations, contributing to the pathogenesis of hereditary DCM can be found in various genes, especially those for sarcolemma elements, contractile elements, Z-disc elements, sarcoplasmic elements, and nuclear lamina elements of cardiomyocytes. On the other hand, disease-associated polymorphisms, which control the susceptibility to non-hereditary DCM, may be found in genes expressing not only in cardiomyocytes but also other non-cardiac cells involved in the immune system. Because functional alterations caused by these genetic variations can be classified into several categories, it is necessary to understand the pathogenesis and hence to develop diagnostic and therapeutic strategies for both hereditary and non hereditary DCM from the viewpoint of genetic factors. PMID- 21617320 TI - Different characteristics of peripartum cardiomyopathy between patients complicated with and without hypertensive disorders. -Results from the Japanese Nationwide survey of peripartum cardiomyopathy-. AB - BACKGROUND: There has been no nationwide survey concerning peripartum cardiomyopathy (PPCM) among the Asian population, and clinical profiles of PPCM complicated with hypertensive disorders complicating pregnancy (HD) as the major risk factor of PPCM have not been characterized. METHODS AND RESULTS: A retrospective, nationwide survey of PPCM in 2007 and 2008 all over Japan was performed and the clinical characteristics were compared between patients with and without HD. We obtained data for 102 patients. HD during pregnancy occurred in 42 patients (41%). Patients with HD were older than those without HD (33.8 vs. 31.9 years old, P<0.05) and babies were delivered more frequently by Caesarean section (81% vs. 52%, P<0.01). Although cardiac parameters at diagnosis were similar in patients with and without HD, patients with HD were hospitalized for a shorter period and had better cardiac function after 7 months. Multivariate regression analysis revealed that HD was independently associated with a shorter hospital stay and a higher left ventricular ejection fraction at last follow up. CONCLUSIONS: PPCM complicated with HD had different clinical characteristics from those without HD. This condition might be a unique subset of PPCM that is characterized by relatively swift recovery except in the cases of death. In order to prevent severe heart failure and maternal death, peripartum women should be treated with HD cautiously and must immediately undergo a cardiac examination as needed. PMID- 21617321 TI - Drug-eluting stent: importance of clinico-pathological correlations. AB - The benefit of drug-eluting stents (DES) is the remarkable reduction in the rates of both restenosis and target lesion revascularization. However, the risk of thrombotic complications extends further in DES-implanted arteries compared with those treated with bare-metal stents (BMS). Moreover, in-stent thrombosis (IST) and delayed arterial healing in DES-treated arteries have been identified by histological examination. At autopsy, proliferation of a monolayer composed of endothelium-like cells over stent struts in DES receiving arteries has been observed; however, these cells are negative for well-accepted endothelial cell markers. An inflammatory reaction against the stent struts is apparent after implantation of BMS and paclitaxel-eluting stents, whereas after sirolimus eluting stents (SES), minimal inflammation is seen up to 6 months after device implantation. IST and in-stent restenosis, both possibly related to a hypersensitivity phenomena, are peculiar to DES, albeit relatively infrequent. A case of enhanced neointimal hyperplasia at 6 months after SES implantation with massive inflammatory reaction including eosinophils, and fibrin deposition is reported here. Observation of the morphological alterations after DES implantation by imaging techniques may furnish important information, but lack of precise comparative data between vascular imaging and histopathology leads to improper interpretation of imaging. Ex vivo imaging using angioscopy, intravascular ultrasound, and optical coherency tomography of SES implantation is presented and the images are compared with the corresponding pathological section. PMID- 21617322 TI - Serum concentration of heart-type fatty acid-binding protein in children and adolescents with congenital heart disease. AB - BACKGROUND: Serum heart-type fatty acid-binding protein (H-FABP) is widely applied as a marker of cardiac myocyte injury. Recently, it has been reported that levels of H-FABP are elevated in adult patients with chronic heart failure and thus provide useful prognostic information. The aim of the present study was to examine the relationships between serum H-FABP levels and pathophysiological characteristics in children and adolescents with congenital heart disease (CHD). METHODS AND RESULTS: Serum H-FABP levels were preoperatively and postoperatively measured in 238 consecutive patients with CHD aged 1-31 years. The relationships between H-FABP levels and severity of heart failure, circulatory status and laboratory data were cross-sectionally analyzed. Multivariate regression analysis indicated that serum H-FABP levels are independently affected by age, New York Heart Association functional class, creatine kinase MB, creatinine and arterial oxygen saturation (standard regression coefficients, -0.378, 0.237, 0.422, 0.615, and -0.210, respectively). Neither left ventricular ejection fraction nor B-type natriuretic peptide correlated with H-FABP levels. CONCLUSIONS: H-FABP could serve as a new monitoring tool to provide information that will guide the optimal therapy and management of CHD patients. PMID- 21617323 TI - Percutaneous valved stent implantation above the coronary ostia. AB - BACKGROUND: In recent years, some experimental and clinical studies on transcatheter aortic valve implantation (TAVI) have been conducted. TAVI is indicated in patients with calcified pure or predominant aortic stenosis. The risk of this technique is still high. Aortic valved stent implantation above the coronary ostia might avoid blocking the coronary ostia. METHODS AND RESULTS: Twenty healthy dogs were selected to establish a canine model of acute aortic valve rupture. The dogs were randomly divided into 2 groups: the rupture model group without any treatment and the valved stent group with percutaneous valved stent implantation above the coronary ostia. The 2 groups of animals were followed up for 3 months. Echocardiography and other tests were performed to assess aortic regurgitation and ventricular function. Acute aortic valve rupture models were successfully established in 16 of 20 dogs. In the rupture model group, the mean aortic regurgitation was 6.8 +/- 1.9 ml/s; only 3 of 8 animals survived for 3 months. In the valved stent group, the mean aortic regurgitation was 7.0 +/- 2.1 ml/s; valved stents were successfully implanted in 8 animals. Instant post-implantation anatomy showed that the stents were located appropriately. Seven dogs survived for 3 months. CONCLUSIONS: Percutaneous valved stent implantation above the coronary ostia is feasible and effective as a transitional treatment for acute aortic valve rupture. PMID- 21617324 TI - Calcium channel antagonists prevent cardiovascular events in Japanese patients with myocardial infarction even in the absence of coronary artery spasm. PMID- 21617325 TI - Transcatheter closure of patent foramen ovale in Chinese patients with paradoxical embolism. -Immediate results and long-term follow-up-. AB - BACKGROUND: The aim of the present study was to assess immediate and long-term clinical outcome of Chinese patent foramen ovale (PFO) patients with paradoxical embolism who underwent transcatheter PFO closure. METHODS AND RESULTS: One hundred and ninety-two patients underwent transcatheter PFO closure for secondary prevention of thromboembolic events (TE). During the procedure, 7 patients had frequent atrial premature beats or transient atrial tachycardia in implantation and 1 patient had a transitory ST-elevation in leads II, III and aV(F). These complications converted spontaneously after a few minutes. No cases of procedure related death or TE were observed during hospitalization. Minor adverse events, including chest discomfort (11%), palpitations (25%) and dyspnea (1%) were reported within 1 month of the procedure. These symptoms had disappeared in most patients by 6-month follow-up. One patient had a new occurrence of migraine at 27 months after the implantation. Within a median follow-up of 49 +/- 8 months, no residual shunt of the atrial level was identified and correct positioning of the device was confirmed on transthoracic echocardiography in all patients. No death related to any cause or recurrent TE were recorded. CONCLUSIONS: Transcatheter PFO closure is a minimally invasive procedure with a high success rate, low complication rate and an excellent long-term outcome, and appears to be a wise approach for secondary prevention of recurrent embolic events in symptomatic patients. PMID- 21617326 TI - Long-term follow-up of patients with isolated left ventricular noncompaction: role of electrocardiography in predicting poor outcome. AB - BACKGROUND: Abnormal baseline electrocardiograms (ECGs) are common in patients with isolated left ventricular noncompaction (IVNC). Whether certain electrocardiographic parameters are associated with a poor clinical outcome, however, remains elusive. The present study was therefore designed to comprehensively assess the predictive value of baseline ECG findings in patients newly diagnosed with IVNC. METHODS AND RESULTS: 74 patients diagnosed with IVNC were included in the analysis. During follow-up, 8 patients (11%) died of a cardiovascular cause or underwent heart transplantation (primary outcome measure). On univariate analysis, several variables, including repolarization abnormalities (ST segment elevation/depression, T-wave inversion) in the inferior leads (5-year estimator: 67.1 +/- 10.7% vs. 98 +/- 2.2%; P = 0.001), an increase in PQ- (hazard ratio (HR) 1.032, P=0.004) and QTc-duration (HR 1.037, P=0.001), were predictive of cardiovascular death or heart transplantation. On multivariate analysis, only PQ- and QTc-duration and the presence of repolarization abnormalities in the inferior leads remained significantly predictive of a poor outcome. CONCLUSIONS: PQ duration, QTc duration, and repolarization abnormalities in the inferior leads are independently predictive of a poor prognosis in IVNC. Further prospective studies are required to conclusively investigate the usefulness of baseline ECG parameters for risk stratification in patients with IVNC. PMID- 21617327 TI - Early development of hyponatremia implicates short- and long-term outcomes in ST elevation acute myocardial infarction. AB - BACKGROUND: Clinical importance of hyponatremia in ST-elevation acute myocardial infarction (STEMI) in the era of primary intervention has not been fully understood. The aim of this study was to investigate the impact of hyponatremia on outcomes in patients with STEMI and secondarily to investigate the contribution of arginine vasopressin (AVP) to hyponatremia in STEMI. METHODS AND RESULTS: Hyponatremia was defined as a sodium concentration <136 mmol/L at 72h after hospitalization. First, the short-term (in-hospital mortality or congestive heart failure (CHF)) and long-term prognosis (cardiac death, re-admission for CHF) in STEMI patients was conducted. Second, the relationship between serum sodium level and plasma AVP was investigated. In hyponatremic patients the incidence of in-hospital heart failure was significantly greater (P=0.0018), long term cardiac death was a higher trend (17.2% vs. 6.3%, P=0.19) and re-admission due to CHF was significantly more frequent (20.7% vs. 4.5%, P=0.0024). Plasma AVP level was higher in the hyponatremia group (4.5 vs. 2.7 pg/ml, P=0.003), and it had a negative correlation with serum sodium level (r=-0.28, P=0.02). CONCLUSIONS: Hyponatremia was frequently found in the early phase of STEMI, and associated with heart failure in both short- and long-term outcomes. Non-osmotic secretion of AVP could be involved in hyponatremia in STEMI patients. PMID- 21617328 TI - Benign cystic nodules may have ultrasonographic features mimicking papillary thyroid carcinoma during interval changes. AB - It had been observed that some cystic nodules change morphologically with ultrasonographic (US) features suspicious for malignancy. The aim of this study was to evaluate the US characteristics of benign cystic nodules mimicking papillary thyroid carcinoma (PTC) during interval changes. Between January 2009 and October 2009, 26 patients with benign cystic nodules showing marked hypoechogenicity in US during the follow-up period were enrolled. During the same period, 38 patients with marked hypoechogenicity in US were enrolled for the PTC group. We evaluated the differences in US characteristics between the 2 groups. Nodule size, margin, echogenic dot and vascularity were not significantly different between the 2 groups. Nodule shape was significantly different between the 2 groups with a lower prevalence of taller than wide in the benign cystic group (11.5% vs. 39.5%, P=0.022). Other coexisting cystic nodules were more frequently observed in benign cystic group (48.3% vs. 5.3%, P=0.001). If echogenic dot was detected in benign cystic nodule, it was more than 1 mm in size without posterior acoustic shadowing unlike echogenic dots in the PTC group. In conclusion, some of the benign cystic nodules may have suspicious malignant features on US during interval changes. A careful assessment of US findings and a previous history may be of value in discriminating them from PTC. PMID- 21617329 TI - Ghrelin in small intestine, its contribution to regulation of food intake and body weight in cross-intestinal parabiotic rats. AB - Ghrelin has been shown to be associated with feeding behavior in humans and rodents. It has been suggested that ghrelin may play a role behind the effect of bariatric surgery. Inbred rats were made into parabiotic pairs so that they shared a single abdominal cavity. A further operation is performed later in which the small intestines are transected and re-connected so that one rat continually lost nutrition to its partner. Changes in food intake and body weight were recorded. Seven weeks later, content of ghrelin in the plasma, stomach and upper intestines were measured in the paired rats. Rats which lost nutrients to its counterpart (Loss rats) ingested significantly more food than sham control rats (p<0.001). Rats which gained nutrient (Gain rats) ingested less than controls (p<0.001). There was no significant difference in body weight, blood glucose, insulin, free fatty acids and triglycerides between the paired rats. There was significantly higher levels of ghrelin in the plasma (p<0.008) and the intestine of the Loss rats (p<0.02). There were no difference in ghrelin in the stomach between parabiotic rats and sham operated controls. The ghrelin content of the plasma and intestines were significantly higher in the Loss rats, which ate more, and normal in the Gain rats, which ate less than controls. Because no remarkable changes in the ghrelin content were observed in the stomach, difference in the quality of the chime may affect the local synthesis and release of ghrelin. PMID- 21617330 TI - The use of plaque score measurements to assess changes in atherosclerotic plaque burden induced by lipid-lowering therapy over time: the METEOR study. AB - AIM: To evaluate whether plaque scoring measurements are able to track changes in atherosclerotic plaque burden over time and to study whether this is affected by lipid-lowering therapy. METHODS: Data used were from METEOR (Measuring Effects on Intima-Media Thickness: an Evaluation Of Rosuvastatin), a randomized controlled trial of rosuvastatin 40 mg among 984 low-risk patients with modest carotid intima-media thickening (CIMT). In this analysis, duplicate ultrasound images from 12 carotid sites were collected at the baseline and end of the study from 495 European patients and were evaluated for plaque presence and severity. Plaques were scored from near and far walls of the 12 sites (0= none; 1= minimal; 2= moderate; 3= severe) and plaque scores (PS) were combined into two summary measures for each examination. The MeanMaxPS is the mean over the 12 carotid sites of the maximum score at each site and the MaxMaxPS reflects the most severe lesion at any site. RESULTS: Baseline MeanMaxPS and MaxMaxPS were 0.31 (SD: 0.20) and 1.15 (SD: 0.51), respectively. Changes in MeanMaxPS and MaxMaxPS significantly differed between rosuvastatin and placebo (mean difference: -0.03 [SE: 0.01; p =0.016] and -0.09 [SE: 0.04; p =0.027], respectively). In contrast to rosuvastatin, which demonstrated no change from the baseline, placebo showed significant progression in MeanMaxPS and MaxMaxPS (p =0.002; both). CONCLUSION: The plaque-scoring method proved capable of assessing the change in atherosclerotic plaque burden over time and proved useful to evaluate lipid lowering in asymptomatic individuals with a low risk of cardiovascular disease and subclinical atherosclerosis. PMID- 21617331 TI - Orally administered rutin inhibits the gene expression of Th2 cytokines in the gut and lung in aged mice. AB - Rutin is one of the flavonoids derived from plants such as buckwheat and is well known as a powerful antioxidant. To determine whether dietary rutin could modulate mucosal immunity, we examined the gene expression of Th1/Th2 cytokines and the receptors in the gut and lung. Aged (18 months old, 18 M) C3H/HeN female mice were orally administered rutin for 10 days. The small intestine and lung were taken and analyzed by real-time PCR for gene expression. Interleukin (IL)-13 and IL-13Ralpha2 gene expression was significantly low (P<0.05 respectively) in the small intestine of aged rutin-fed mice. Meanwhile, there was no change in interferon gamma gene expression between control and rutin-fed mice. IL-13 gene expression was also downregulated in the lung. To examine the mechanism of the inhibitory effect of rutin on Th2 cytokines in aged mice, intestinal nitric oxide synthase (NOS) expression was evaluated. Rutin inhibited inducible NOS (NOS2) gene expression, but not neuronal NOS and endothelial NOS. Gene analysis of cells collected from the small intestine by laser capture dissection revealed that NOS2 expression was significantly inhibited in crypt regions. Thus, rutin might be effective against a Th2-dominant profile through NOS2 inhibition in aged mice. PMID- 21617332 TI - Fibrolipoma of a cryptorchid testis in a young bull. AB - A six-month-old Japanese Black bull was found to have no left testis in the scrotum. A fist-sized mass was palpated per rectum. Two months later, hCG was injected and blood samples were collected before and after injection. No testosterone response to hCG was observed. On the cut surface of the excised mass, most of the mass was composed of homogeneous adipose-like tissue. The rest of the surface was composed of a well-circumscribed testicular parenchyma-like tissue (18 * 16 * 15 mm). Histology revealed diffusely distributed mature adipocytes and septa of fibrous connective tissue. Neither germ cells nor spermatozoa were observed in the seminiferous tubules. A diagnosis was made of fibrolipoma and aspermatogenesis of the left cryptorchid testis. PMID- 21617333 TI - Short sleep duration and irregular bedtime are associated with increased behavioral problems among Japanese preschool-age children. AB - Sleep problems are known to be risk factors for subsequent emotional and behavioral difficulties in childhood and adolescence. To date, there has been no study investigating the relationships between sleep habits and behavioral problems in a large nonclinical sample of preschool age children. The aim of this study was to examine these relationships and factors associated with the sleep habits of preschool age (2 to 5 year old) children. Their mothers (n = 1,746) completed a multiple-choice questionnaire about the sleep habits and behavior problems of their children, as well as their own sleep habits and working hours at Tokyo metropolitan public nursery schools. The short sleep duration group showed significantly higher aggressive scores than the long sleep duration group among 2- to 3-year-old children, and the irregular bedtime group showed significantly higher aggressive and attention problem scores than the regular bedtime group among 4- to 5-year-old children. Univariate and multivariate logistic regression analyses revealed that children's late bedtime was associated with their mother's late waking-up time, and late schedule of both the mother's leaving and returning home. This study recognized an association between behavioral problems and poor sleep habits among preschool-age children. It is important for children to sleep regularly and adequately in order to decrease their behavior problems. In conclusion, appropriate management of children's sleep by their mothers is necessary for promoting sleep-related health of children. PMID- 21617334 TI - Seasonal variation in estimated glomerular filtration rate based on serum creatinine levels in hypertensive patients. AB - Seasonal variations in blood pressures should be kept in mind when controlling blood pressure in hypertensive patients. Seasonal variations in glomerular filtration rate (GFR) also may have a clinical significance. However, it is time consuming to measure GFR directly. We therefore examined the seasonal variation in estimated glomerular filtration rate (eGFR) based on serum creatinine levels in hypertensive patients without CKD (eGFR >= 60 mL/min/1.73 m(2)) and those with chronic kidney disease (CKD) (eGFR < 60 mL/min/1.73 m(2)). This study included 47 hypertensive patients without CKD (69 +/- 11 yrs) and 55 hypertensive patients with CKD (76 +/- 8 yrs). The eGFR was determined from the equation: eGFR = 194 * age(-0.287) * (serum creatinine)(-1.094) (* 0.739 if female). Overall, both groups of hypertensive patients demonstrated similar seasonal variations in eGFR. Importantly, hypertensive patients without CKD and those with CKD showed the lower eGFR in summer (June-August) (71.8 +/- 13.2 and 37.2 +/- 13.0 mL/min/1.73 m(2), respectively) compared with the eGFR in spring (March-May) (77.9 +/- 13.0 and 43.0 +/- 14.0 mL/min/1.73 m(2), respectively) (p < 0.05). The decrease in eGFR from spring to summer was similar for both types of hypertensive patients (without CKD, -6.1 +/- 7.0; with CKD, -5.8 +/- 5.2 mL/min/1.73 m(2)). However, the percent change in eGFR from spring to summer was greater in hypertensive patients with CKD (-13.8 +/- 9.4 %) than in those without CKD (-7.7 +/- 8.3 %) (p = 0.001). In conclusion, careful observation regarding renal function is needed for hypertensive patients with CKD during summer. PMID- 21617335 TI - Cost effectiveness of interventions to promote screening for colorectal cancer: a randomized trial. AB - OBJECTIVES: Screening for colorectal cancer is considered cost effective, but is underutilized in the U.S. Information on the efficiency of "tailored interventions" to promote colorectal cancer screening in primary care settings is limited. The paper reports the results of a cost effectiveness analysis that compared a survey-only control group to a Centers for Disease Control (CDC) web based intervention (screen for life) and to a tailored interactive computer-based intervention. METHODS: A randomized controlled trial of people 50 and over, was conducted to test the interventions. The sample was 1224 participants 50-70 years of age, recruited from Kelsey-Seybold Clinic, a large multi-specialty clinic in Houston, Texas. Screening status was obtained by medical chart review after a 12 month follow-up period. An "intention to treat" analysis and micro costing from the patient and provider perspectives were used to estimate the costs and effects. Analysis of statistical uncertainty was conducted using nonparametric bootstrapping. RESULTS: The estimated cost of implementing the web-based intervention was $40 per person and the cost of the tailored intervention was $45 per person. The additional cost per person screened for the web-based intervention compared to no intervention was $2602 and the tailored intervention was no more effective than the web-based strategy. CONCLUSIONS: The tailored intervention was less cost-effective than the web-based intervention for colorectal cancer screening promotion. The web-based intervention was less cost effective than previous studies of in-reach colorectal cancer screening promotion. Researchers need to continue developing and evaluating the effectiveness and cost-effectiveness of interventions to increase colorectal cancer screening. PMID- 21617336 TI - Changes in the distribution of maternal age and parity and increasing trends in the low birth weight rate in Korea between 1995 and 2005. AB - OBJECTIVES: This study measured the impact of shifts in maternal age and parity on the increase in the low birth weight (LBW) rate in Korea. METHODS: We obtained raw data for all 6 397 945 live births registered at the Korea National Statistical Office between 1995 and 2005. We calculated the proportion of increment in the LBW rate due to changes in the distribution of maternal age and parity (AP-dis) and the proportion due to changes in the age- and parity-specific LBW rate (AP-spe). RESULTS: The LBW rate increased from 3.02% in 1995 to 4.28% in 2005. The multiple birth rate increased from 1.32% to 2.19% during the same period. Of the 1.26% points increment in the LBW rate, 0.64% points occurred among singleton births and 0.62% points occurred among multiple births. Changes in the AP-dis accounted for 50% of the increase in the LBW rate among singleton births, but did not contribute to the increase in the LBW rate among multiple births. The remainder of the total increment in the LBW rate was explained by the increase in the AP-spe. CONCLUSIONS: This study demonstrated that shifts in maternal age and parity among singleton births and increased multiple births were important contributors to the increment in the LBW rate. This study also revealed that the increase in the AP-spe was an equally important contributor as the shifts in maternal age and parity to the increment in the LBW rate among singleton births and was a major contributor among multiple births. PMID- 21617337 TI - Modifying effect of suicidal ideation on the relationship between asthma and cigarette use behaviors among Korean adolescents. AB - OBJECTIVES: Although cigarette smoking is known to be related to the exacerbation of asthma symptoms, several studies have indicated that the prevalence of cigarette smoking among asthmatic adolescents is similar to or even higher than that among non-asthmatic adolescents. The aim of this study was to evaluate the relationship between asthma and cigarette use behaviors and whether or not the presence of suicidal ideation modifies this relationship among Korean adolescents. METHODS: We analyzed data from the 2008 Korea Youth Risk Behavior Web-based Survey, which included a nationally representative sample of middle and high school students. Multiple logistic regression models were used to calculate odd ratios and 95% confidence intervals of cigarette use behaviors among current asthmatics, former asthmatics, and non-asthmatics, after adjusting for gender, grade, school records, socioeconomic status, current alcohol use, and suicidal ideation. RESULTS: Of 75 238 study participants, 3.5% were current asthmatics and 4.5% were former asthmatics. Compared with non-asthmatics, asthmatics were more likely to report current cigarette use, frequent and heavy cigarette use, and cigarette use before 13 years of age. There were statistically significant interactions between asthma and suicidal ideation in cigarette use behaviors. CONCLUSIONS: This study demonstrated that asthmatic adolescents are more likely than non-asthmatic adolescents to engage in cigarette use behaviors and the presence of suicidal ideation is an effect modifier of the relationship between asthma and cigarette use behaviors. Particular attention should be paid to the awareness of health risks of cigarette smoking and mental health problems among asthmatic adolescents. PMID- 21617338 TI - Association between microRNA196a2 rs11614913 genotypes and the risk of non-small cell lung cancer in Korean population. AB - OBJECTIVES: The microRNA (miRNA) miR-196a2 may play an important role in lung cancer development and survival by altering binding activity of target mRNA. In this study, we evaluated their associations with the susceptibility of non-small cell lung cancers (NSCLC) by case-control study in a Korean population. METHODS: We performed genotyping analyses for miR-196a2 rs11614913 T/C at miRNA regions in a case-control study using blood samples of 406 NSCLC patient and 428 cancer-free control groups. RESULTS: The total C allele frequencies for miR-196a2 were 48.8% for the patients and 45.6% for the controls; and the genotype frequencies of TT, TC, and CC were 23.7%, 55.2%, and 21.1% for the patients and 31.1%, 46.35%, and 22.4% for the controls (p<0.05). Participants who possesses TC/CC genotypes showed high risk for NSCLC compared to those possessed TT genotypes (OR, 1.42; 95% CI, 1.03 to 1.96). The association was persisted in 60 and older age group, male, smokers, those without family history for cancer. However, no significant association of CC genotypes in recessive genetic model was observed. CONCLUSIONS: In conclusion, this case-control study provides evidence that miR-196a2 rs11614913 C/T polymorphisms are associated with a significantly increased risk of NSCLC in a dominant model, indicating that common genetic polymorphisms in miR 196a2 rs11614913 are associated with NSCLC. The association of miR196a2 rs11614913 polymorphisms and NSCLC risk require confirmation through additional larger studies. PMID- 21617339 TI - Frequency and related factors of masked hypertension at a worksite in Korea. AB - OBJECTIVES: Masked hypertension is associated with metabolic risks and increased risk of cardiovascular disease. The purpose of this study was to identify the frequency of and risk factors of masked hypertension in Korean workers. METHODS: The study was conducted among 121 employees at a hotel in Gyeongju, Korea, from December 2008 to February 2009. We measured blood pressure (BP) both in the clinic and using 24-hour ambulatory BP monitors for all subjects. Hypertension was defined independently by both methods, and subjects were classified into four groups: true normotension, masked hypertension, white coat hypertension, and sustained hypertension. RESULTS: The frequency of masked hypertension in our study group was 25.6%. Compared with true normotension, the factors related to masked hypertension were male gender (odds ratio [OR], 10.7; 95% confidence interval [CI], 1.41 to 81.09), aging one year (OR, 0.88; 95% CI, 0.78 to 0.99), clinic BP 120-129/80-84 mmHg (OR, 8.42; 95% CI, 1.51 to 46.82), clinic BP 130-139 / 85-89 mmHg (OR, 12.14; 95% CI, 1.80 to 81.85), smoking (OR, 5.51; 95% CI, 1.15 to 26.54), and increase of total cholesterol 1 mg / dL (OR, 1.05; 95% CI, 1.02 to 1.08). In males only, these factors were clinic BP 120-129 / 80-84 mmHg (OR, 15.07; 95% CI, 1.55 to 146.19), clinic BP 130-139 / 85-89 mmHg (OR, 17.16; 95% CI, 1.56 to 189.45), smoking (OR, 11.61; 95% CI, 1.52 to 88.62), and increase of total cholesterol 1 mg/dL (OR, 1.05; 95% CI, 1.01 to 1.09). CONCLUSIONS: The frequency of masked hypertension was high in our study sample. Detection and management of masked hypertension, a known strong predictor of cardiovascular risk, could improve prognosis for at-risk populations. PMID- 21617340 TI - Study of specific oligosaccharide structures related with swine flu (H1N1) and avian flu, and tamiflu as their remedy. AB - The infection of pandemic influenza viruses such as swine flu (H1N1) and avian flu viruses to the host cells is related to the following two factors: First, the surface protein such as HA (hemagglutinin) and NA (neuraminidase) of the influenza virus. Second, the specific structure of the oligosaccharide [sialic acid(alpha2-6) galactose(beta1-4)glucose or sialic acid(alpha2-3)galactose(beta1 4)glucose] on the host cell. After recognizing the specific structure of the oligosaccharide on the surface of host cells by the surface protein of the influenza virus, the influenza virus can secrete sialidase and cleave the sialic acid attached on the final position of the specific structure of the oligosaccharide on the surface of host cells. Tamiflu (oseltamivir), known as a remedy of swine flu, has a saccharide analog structure, especially the sialic acid analog. Tamiflu can inhibit the invasion of influenza viruses (swine flu and avian flu viruses) into the host cells by competition with sialic acid on the terminal position of the specific oligosaccharide on the surface of the host cell. Because of the emergence of Tamiflu resistance, the development of new potent anti-influenza inhibitors is needed. The inhibitors with positive-charge groups have potential as antiviral therapeutics, and the strain specificity must also be resolved. PMID- 21617341 TI - Genome organization and transcription response to harvest of two metallothionein like genes in Agaricus bisporus fruiting bodies. AB - Metallothioneins are a class of small cysteine-rich proteins that have been associated with increased tolerance to metal and oxidative stresses in animals, plants, and fungi. We investigated a metallothionein-like (mt-like) gene shown previously to be upregulated in fruiting bodies of the fungus Agaricus bisporus in response to post-harvest storage. Analysis of an A. bisporus genomic DNA cosmid library identified two similar mt-like genes (met1 and met2) arranged as a bidirectional gene pair transcribed from the same promoter region. The promoter contained regulatory elements including 9 metal responsive elements and a CAAT box region 220 bp downstream of met1 that showed striking similarity to a feature in Coprinopsis cinerea mt-like gene promoters. Transcriptional analysis showed that both met genes are significantly and rapidly (within 3 hours) upregulated during post-harvest storage and expression is significantly greater in stipe and cap tissues compared with the gills. However, a strong directionality of the promoter was demonstrated, as transcript levels of met1 were at least two orders of magnitude greater than those of met2 in all samples tested. PMID- 21617342 TI - Ferric reductase activity of the ArsH protein from Acidithiobacillus ferrooxidans. AB - The arsH gene is one of the arsenic resistance system in bacteria and eukaryotes. The ArsH protein was annotated as a NADPH-dependent flavin mononucleotide (FMN) reductase with unknown biological function. Here we report for the first time that the ArsH protein showed high ferric reductase activity. Glu104 was an essential residue for maintaining the stability of the FMN cofactor. The ArsH protein may perform an important role for cytosolic ferric iron assimilation in vivo. PMID- 21617343 TI - Isolation, optimization, and partial purification of amylase from Chrysosporium asperatum by submerged fermentation. AB - A potent fungus for amylase production, Chrysosporium asperatum, was isolated from among 30 different cultures obtained from wood samples collected in the Junagadh forest, India. All of the isolated cultures were screened for their ability to produce amylase by submerged fermentation. Among the selected cultures, C. asperatum (Class Euascomycetes; Onygenales; Onygenaceae) gave maximum amylase production. In all of the different media tested, potato starch was found to be a good substrate for production of amylase enzyme at 30 degrees C and pH 5.0. Production of enzyme reached the maximum when a combination of starch and 2% xylose, and organic nitrogen (1% yeast extract) and ammonium sulfate were used as carbon and nitrogen sources, respectively. There was no significant effect of metal ions on enzyme activity. The enzyme was relatively stable at 50 degrees C for 20 min, and no inhibitory effect of Ca+2 ions on amylase production was observed. PMID- 21617344 TI - Growth-suppressing activity of the transfected Cx26 on BICR-M1Rk breast cancer cell line. AB - There are accumulating evidences suggesting that connexin (Cx), a gap junction channel-forming protein, acts as a growth suppressor in various cancer cells, and this effect is attributed to the gap junction-mediated intercellular communication (GJIC). In order to characterize the relationship between the growth-arresting activity of Cx26 and its cytoplasmic localizations after expression, we linked a nuclear export signal (NES) sequence to Cx26 cDNA before transfecting into a rat breast cancer cell line. A confocal fluorescent microscopic observation revealed that the insertion of NES minimized the nuclear expression of Cx26, and increased its cytoplasmic expression, including plasma membrane junctions. Total cell counting and BrdUrd-labeling experiments showed that the growth of the breast cancer cells was inhibited by 74% upon transfection of Cx26-NES, whereas only 9% inhibition was observed with only Cx26 cDNA. PMID- 21617345 TI - Identification and characterization of the acid phosphatase HppA in Helicobacter pylori. AB - An acid phosphatase (HppA) activated by NH4Cl was purified 192- and 34-fold from the periplasmic and membrane fractions of Helicobacter pylori, respectively. SDS polyacrylamide gel electrophoresis revealed that HppA from the latter appears to be several kilodaltons larger in molecular mass than from the former by about 24 kDa. Under acidic conditions (pH< or =4.5), the enzyme activity was entirely dependent on the presence of certain mono- and/or divalent metal cations (e.g., K+, NH4 +, and/or Ni2+). In particular, Ni2+ appeared to lower the enzyme's Km for the substrates, without changing Vmax. The purified enzyme showed differential specificity against nucleotide substrates with pH; for example, the enzyme hydrolyzed adenosine nucleotides more rapidly at pH 5.5 than at pH 6.0, and vice versa for CTP or TTP. Analyses of the enzyme's N-terminal sequence and of an HppA- H. pylori mutant revealed that the purified enzyme is identical to rHppA, a cloned H. pylori class C acid phosphatase, and shown to be the sole bacterial 5'-nucleotidase uniquely activated by NH4Cl. In contrast to wild type, HppA- H. pylori cells grew more slowly. Strikingly, they imported Mg2+ at a markedly lowered rate, but assimilated urea rapidly, with a subsequent increase in extracellular pH. Moreover, mutant cells were much more sensitive to extracellular potassium ions, as well as to metronidazole, omeprazole, or thiophenol, with considerably lowered MIC values, than wild-type cells. From these data, we suggest that the role of the acid phosphatase HppA in H. pylori may extend beyond 5'-nucleotidase function to include cation-flux as well as pH regulation on the cell envelope. PMID- 21617346 TI - Coenzyme Q10 production by Sphingomonas sp. ZUTE03 with novel precursors isolated from tobacco waste in a two-phase conversion system. AB - Coenzyme Q10 (CoQ10) is a widely used supplement in heart diseases treatment or antioxidative dietary. The microbial production of CoQ10 was enhanced by addition of solanesol and novel precursors recovered from waste tobacco. The novel precursors were separated by silica gel and identified as alpha-linolenic acid (LNA) and butylated hydroxytoluene (BHT) based on the effect on CoQ10 production and GC-MS. The effects of novel precursors on CoQ10 production by Sphingomonas sp. ZUTE03 were further evaluated in a two-phase conversion system. The precursor's combination of solanesol (70 mg/l) with BHT (30 mg/l) showed the best effect on the improvement of CoQ10 yield. A maximal CoQ10 productivity (9.5 mg l 1 h-1) was achieved after 8 h conversion, with a molar conversion rate of 92.6% and 92.4% on BHT and solanesol, respectively. The novel precursors, BHT and LNA in crude extracts from waste tobacco leaves, might become potential candidates for application in the industrial production of CoQ10 by microbes. PMID- 21617347 TI - Purification and characterization of an extracellular beta-glucosidase produced by Phoma sp. KCTC11825BP isolated from rotten mandarin peel. AB - A beta-glucosidase from Phoma sp. KCTC11825BP isolated from rotten mandarin peel was purified 8.5-fold with a specific activity of 84.5 U/mg protein. The purified enzyme had a molecular mass of 440 kDa with a subunit of 110 kDa. The partial amino acid sequence of the purified beta-glucosidase evidenced high homology with the fungal beta- glucosidases belonging to glycosyl hydrolase family 3. Its optimal activity was detected at pH 4.5 and 60 degrees C, and the enzyme had a half-life of 53 h at 60 degrees C. The Km values for p-nitrophenyl-beta-D glucopyranoside and cellobiose were 0.3 mM and 3.2 mM, respectively. The enzyme was competitively inhibited by both glucose (Ki=1.7 mM) and glucono-delta-lactone (Ki=0.1 mM) when pNPG was used as the substrate. Its activity was inhibited by 41% by 10 mM Cu2+ and stimulated by 20% by 10 mM Mg2+. PMID- 21617348 TI - The phenotypic and genotypic characterization of Korean isolates of Cronobacter spp. (Enterobacter sakazakii). AB - This study was conducted to investigate the phenotypic and genotypic characteristics of Korean isolates of Cronobacter spp. (Enterobacter sakazakii). A total of 43 Cronobacter spp., including 5 clinical isolates, 34 food isolates, 2 environmental isolates, and 2 reference strains (C. sakazakii ATCC 29004 and C. muytjensii ATCC51329) were used in this study. Korean isolates of Cronobacter spp. were divided into 11 biogroups according to their biochemical profiles and 3 genomic groups based on the analysis of their 16S rRNA gene sequences. Biogroups 1 and 2 contained the majority of isolates (n=26), most of which were contained in 16S rRNA cluster 1 (n=34). Korean isolates of Cronobacter spp. showed diverse biochemical profiles. Biogroup 1 contained C. sakazakii GIHE (Gyeonggido Research Institute of Health and Environment) 1 and 2, which were isolated from babies that exhibited symptoms of Cronobacter spp. infection such as gastroenteritis, sepsis, and meningitis. Our finding revealed that Biogroup 1, C. sakazakii, is more prevalent and may be a more pathogenic biogroup than other biogroups, but the pathogenic biogroup was not represented clearly among the 11 biogroups tested in this study. Thus, all biogroups of Cronobacter spp. were recognized as pathogenic bacteria, and the absence of Cronobacter spp. in infant foods should be constantly regulated to prevent food poisoning and infection caused by Cronobacter spp. PMID- 21617349 TI - Cloning and expression of a bpr gene encoding Bacillopeptidase F from Bacillus amyloliquefaciens CH86-1. AB - A gene encoding bacillopeptidase F, bpr86-1, was cloned from B. amyloliquefaciens CH86-1 isolated from cheonggukjang. This gene could encode a preproenzyme of 1,431 amino acids. When bpr86-1 was introduced into B. subtilis WB600 via pHY300PLK, an E. coli-Bacillus shuttle vector, the transformant showed fibrinolytic activity. During growth on LB, the fibrinolytic activity of cells increased sharply when they entered the stationary phase. The highest activity (761.4 mU/mg protein) was observed at 96 h of cultivation. PMID- 21617350 TI - Impact of fermentation rate changes on potential hydrogen sulfide concentrations in wine. AB - The correlation between alcoholic fermentation rate, measured as carbon dioxide (CO2) evolution, and the rate of hydrogen sulfide (H2S) formation during wine production was investigated. Both rates and the resulting concentration peaks in fermentor headspace H2S were directly impacted by yeast assimilable nitrogenous compounds in the grape juice. A series of model fermentations was conducted in temperature-controlled and stirred fermentors using a complex model juice with defined concentrations of ammonium ions and/or amino acids. The fermentation rate was measured indirectly by noting the weight loss of the fermentor; H2S was quantitatively trapped in realtime using a pre-calibrated H2S detection tube which was inserted into a fermentor gas relief port. Evolution rates for CO2 and H2S as well as the relative ratios between them were calculated. These fermentations confirmed that total sulfide formation was strongly yeast strain dependent, and high concentrations of yeast assimilable nitrogen did not necessarily protect against elevated H2S formation. High initial concentrations of ammonium ions via addition of diammonium phosphate (DAP) caused a higher evolution of H2S when compared with a non-supplemented but nondeficient juice. It was observed that the excess availability of a certain yeast assimilable amino acid, arginine, could result in a more sustained CO2 production rate throughout the wine fermentation. The contribution of yeast assimilable amino acids from conventional commercial yeast foods to lowering of the H2S formation was marginal. PMID- 21617351 TI - RKIP downregulation induces the HBx-mediated Raf-1 mitochondrial translocation. AB - The Raf-1 kinase inhibitory protein (RKIP) can regulate multiple key signaling pathways. Specifically, RKIP binds to Raf-1 kinase and inhibits the Ras-Raf-1 MEK1/2- ERK1/2 pathway. Additionally, Raf-1 has been shown to translocate to mitochondria and thereby protect cells from stress-mediated apoptosis. Recently, HBx was found to stimulate the mitochondrial translocation of Raf-1, contributing to the anti-apoptotic effect. We found that RKIP was downregulated during HBx mediated hepatocarcinogenesis. In this study, we show that RKIP bound to Raf-1 and consequently inhibited the translocation of Raf-1 into mitochondria. This promoted the apoptosis of cells treated with apoptotic stimulus. Thus, the downregulation of RKIP increased the level of free Raf-1 and thereby elevated the mitochondrial translocation of Raf-1 during HBx-mediated hepatocarcinogenesis. The elevated Raf-1 mitochondrial translocation induced the increased anti apoptotic effect and subsequently promoted HBx-mediated hepatocarcinogenesis. PMID- 21617352 TI - Production of a recombinant anti-human CD4 single-chain variable-fragment antibody using phage display technology and its expression in Escherichia coli. AB - Single-chain variable fragment (scFv) is a fusion protein of the variable regions of the heavy (VH) and light (VL) chains of immunoglobulin, connected with a short linker peptide of 10 to about 20 amino acids. In this study, the scFv of a monoclonal antibody against the third domain of human CD4 was cloned from OKT4 hybridoma cells using the phage display technique and produced in E. coli. The expression, production, and purification of anti-CD4 scFv were tested using SDS PAGE and Western blot, and the specificity of anti-CD4 scFv was examined using ELISA. A 31 kDa recombinant anti-CD4 scFv was expressed and produced in bacteria, which was confirmed by SDS-PAGE and Western blot assays. Sequence analysis proved the ScFv structure of the construct. It was able to bind to CD4 in quality ELISA assay. The canonical structure of anti-CD4 scFv antibody was obtained using the SWISS_MODEL bioinformatics tool for comparing with the scFv general structure. To the best of our knowledge, this is the first report for generating scFv against human CD4 antigen. Engineered anti-CD4 scFv could be used in immunological studies, including fluorochrome conjugation, bispecific antibody production, bifunctional protein synthesis, and other genetic engineering manipulations. Since the binding site of our product is domain 3 (D3) of the CD4 molecule and different from the CD4 immunological main domain, including D1 and D2, further studies are needed to evaluate the anti-CD4 scFv potential for diagnostic and therapeutic applications. PMID- 21617353 TI - Colorimetric evaluation of the time-killing assay for Citropin 1.1, lipopeptide Palm-KK-NH2, and Temporin A. AB - Nowadays, there are a number of colorimetric techniques available for the determination of a time killing assay in a manner much easier and faster than those previously more commonly used, which were much more time-consuming and laborious colony counting procedures. Here, an attempt has been made to test the antimicrobial peptides of Citropin 1.1, Palm-KK-NH2, and Temporin A on a reference strain of Staphylococcus aureus using resazurin as the cell viability reagent. Staphylococcus aureus was exposed to the test compounds over varying periods of time and the metabolic activity measured, with a profile of antimicrobial activity then established. The results are in agreement with data from previous literature, thus confirming the relevance of the application of resazurin for the testing of antimicrobial agents. PMID- 21617354 TI - Benzyldihydroxyoctenone, a novel nonsteroidal antiandrogen, shows differential apoptotic induction in prostate cancer cells in response to their androgen responsiveness. AB - The molecular mechanisms of apoptotic induction by benzyldihydroxyoctenone (BDH), a nonsteroidal antiandrogen, isolated from the culture broth of Streptomyces sp., have been previously published in prostate cancer LNCaP cells. Apoptotic induction of BDH-treated LNCaP cells was associated with downregulation of Bcl-xL that caused, in turn, cytochrome c release from mitochondria, and activation of procaspases and specific proteolytic cleavage of poly(ADP-ribose) polymerase (PARP). The purpose of the present study was to investigate the patterns of apoptotic induction by BDH in non-prostate, ovarian cancer PA-1 (androgen independent and -insensitive) cells and prostate cancer cells with different androgen responsiveness, such as C4-2 (androgen-independent and -sensitive), 22Rv1 (androgen-dependent and -low sensitive), and LNCaP (androgen-dependent and high sensitive) cells. We found that BDH-treated LNCaP cell proliferation was significantly inhibited in a time-dependent manner and induced apoptosis via downregulation of the androgen receptor (AR) and prostate-specific antigen (PSA), as well as antiapoptotic Bcl-xL protein. However, the levels of BDH-mediated apoptotic induction and growth inhibition in 22Rv1 cells were apparently lower than those of LNCaP cells. In contrast, the induction of apoptosis and antiproliferative effect in BDH-treated non-prostate cancer PA-1 and hormone refractory C4-2 cells were not detectable and marginal, respectively. Therefore, BDH-mediated differential apoptotic induction and growth inhibition in a cell type seem to be obviously dependent on its androgen responsiveness; primarily on androgen-dependency, and then on androgen sensitivity. PMID- 21617355 TI - [Eosinophilic chronic rhinosinusitis and eosinophilic otitis media]. PMID- 21617356 TI - [Episodic angioedema with eosinophilia]. PMID- 21617357 TI - [Recent advances in guidelines associated with food allergy (how to cope with food allergy patients)]. PMID- 21617358 TI - [Molecular mechanisms for dendritic cell subset functions]. PMID- 21617359 TI - [A multipotential role of the transcriptional repressor BCL6 in the pathogenesis of allergic inflammation]. PMID- 21617360 TI - [Clinical study concerning anti-inflammatory effect of fixed dose therapy by budesonide/formoterol combination inhaler for moderate persistent asthmatics]. AB - BACKGROUND: The clinical usefulness of fixed dose maintenance therapy using a combination inhaler containing budesonide and formoterol (FBC) has already been established, still evidence concerning anti-inflammatory effect by maintenance therapy with fix-dosed FBC, in comparison with other ICS and LABA combination therapy including salmeterol/fluticasone combination inhaler, is lacking. METHODS: Moderate persistent adult asthmatics who has received combination therapy of ICS (200~500 MUg/day FP equivalent) and LABA (salmeterol 100 MUg/day) for more than 6 months and under well-control conditions by asthma control test (ACT) have been recruited. FeNO, as a marker of airway inflammation, ACT score, and the results of spirometry have been evaluated after switching to maintenance therapy by FBC 640/18 MUg/day for 8 weeks. RESULTS: The fixed dosed FBC therapy resulted in superior primary outcome, as compared with previous ICS/LABA combination therapy, as assessed by measuring FeNO, 44.0+/-26.5 to 31.3+/- 15.4 ppb (p<0.01, paired-t test). ACT score also improved significantly, 22.22+/-1.57 to 23.88+/-1.57 (p<0.01). The number of patients who used SABA more than once a week has decreased 13 to 0 with FBC treatment. CONCLUSION: The airway anti inflammatory effect by 8 weeks maintenance therapy with fix-dosed FBC 640/18 MUg/day (2 puffs twice a day) has been strongly suggested. PMID- 21617361 TI - [A trial of hierarchical cluster analysis of hospitalized wheezy infants and young children based on clinical factors]. AB - BACKGROUND: It has been estimated that there are several phenotypes constituting wheezy infants, in addition to true asthma. OBJECTIVE: The aim of this study is categorized wheezy infants and young children by cluster analysis and to turn out of categorized frequency. SUBJECTS AND METHODS: The study included 53 subjects aged less than 2 years old who were admitted with wheezy exacerbation. Based on clinical factors, we classified these subjects into several clinical phenotypes using cluster analysis. RESULTS: The following four phenotypes were identified; cluster 1: almost all were exacerbated by RSV infection without previous wheezy episodes; cluster 2: almost all were younger, males, without previous wheezy episodes, sensitized to allergens and parental asthma, who tended to be exposed to passive smoke and have no siblings; cluster 3: almost all were slightly older males with recurrent wheezy episodes who tended to be sensitized to various allergens; cluster 4: almost all were younger females with exposure to passive smoke, whose mother or father has a history of asthma. Cluster 4 patients tended to attend a day-care center and to be exclusively breast-fed. CONCLUSION: Although this study covers only a limited number of subjects, cluster analysis is a new and useful method of categorizing heterogeneous wheezy infants and young children. Further analysis may establish clinical classifications of these patients. PMID- 21617362 TI - [Factors influencing parental medication adherence in pediatric asthma]. AB - PURPOSE: To identify the factors influencing parental medication control behaviors (inhaling corticosteroids and medication-taking) in pediatric asthma management. METHODS: A specially-designed questionnaire survey was conducted on 942 parents with asthmatic children in hospitals and elementary schools. RESULTS: Factor analysis on inhalation behaviors resulted in five factors: understanding of benefit, mastering on inhalation skills and medication management, family support, anxiety of side effects, and explanation from a doctor (cumulative contribution ratio=51.3%). Factor analysis on medication-taking behavior resulted in five factors: understanding of medication effectiveness and benefit, family support, anxiety of side effects, skills on giving medicines, and family routine (cumulative contribution ratio=50.6%). CONCLUSION: The results indicate the importance of recognizing factors influencing parental medication control behaviors in developing education strategies to maintain and reinforce their asthma management behaviors. PMID- 21617363 TI - [The efficacy of shortening an administration period with omalizumab]. AB - A 64-year old male who had been treated with oral predisolone for severe bronchial asthma had started the treatment of omalizumab administration every 4 weeks since December 2009 and his symptoms had prominently improved. However, he complained the recurrence of his symptoms 4 weeks after administration of omalizumab and the mean times of SABA use had also increased from 0.61/day (1-2w) to 0.95/day (4w). Therefore, we had shortened the interval of omalizumab to 3 from 4 weeks since April 2010. Consequently, his symptoms have improved with increase of Asthma Control Test (15.67->21.33) and it has been possible to reduce the oral prednisolone. Recently, we can use the omalizumab and have extended the choice of treatment for severe bronchial asthma. However, it is predicted that there are some patients who have clinical problems for continuing the recommendation dose or intervals. We report a case with a brief review of the literature. PMID- 21617365 TI - Stem cells for organ repair: support or replace? PMID- 21617366 TI - Chromatin remodeling and the circadian clock: Jumonji C-domain containing proteins. AB - Circadian rhythms are a universal way for organisms, ranging from prokaryotes to humans, to maintain coordination with the daily changes of light and temperature. It is known that a functional circadian clock confers enhanced fitness. In both animals and plants, diverse physiological processes are affected by the clock and more than 10% of transcripts show a circadian rhythm. Recent advances in the field have revealed a link between circadian regulated gene expression and dynamic changes in chromatin. Jumonji C (JmjC) domain-containing proteins have been shown to be involved in chromatin remodeling, acting as histone demethylases. The recent discovery that a JmjC-domain containing protein functions as a novel clock component suggests that histone modification has evolved as an important mechanism at the core of the circadian machinery. PMID- 21617367 TI - A genomic point-of-view on environmental factors influencing the human brain methylome. AB - The etiologic paradigm of complex human disorders such as autism is that genetic and environmental risk factors are independent and additive, but the interactive effects at the epigenetic interface are largely ignored. Genomic technologies have radically changed perspective on the human genome and how the epigenetic interface may impact complex human disorders. Here, I review recent genomic, environmental, and epigenetic findings that suggest a new paradigm of "integrative genomics" in which genetic variation in genomic size may be impacted by dietary and environmental factors that influence the genomic saturation of DNA methylation. Human genomes are highly repetitive, but the interface of large scale genomic differences with environmental factors that alter the DNA methylome such as dietary folate is under-explored. In addition to obvious direct effects of some environmental toxins on the genome by causing chromosomal breaks, non mutagenic toxin exposures correlate with DNA hypomethylation that can lead to rearrangements between repeats or increased retrotransposition. Since human neurodevelopment appears to be particularly sensitive to alterations in epigenetic pathways, a further focus will be on how developing neurons may be particularly impacted by even subtle alterations to DNA methylation and proposing new directions towards understanding the quixotic etiology of autism by integrative genomic approaches. PMID- 21617369 TI - Hypothesis: environmental regulation of 5-hydroxymethylcytosine by oxidative stress. AB - Many environmental toxins, such as heavy metals, air particles, and ozone, induce oxidative stress and decrease the levels of NADH and NADPH, cofactors that drive anabolic biochemical reactions and provide reducing capacity to combat oxidative stress. Recently, it was found that the Ten-eleven translocation (TET) protein family members, which oxidize 5-methylcytosine (5-mC) to 5-hydroxymethylcytosine (5-hmC) in the DNA, is activated under high oxygen conditions by alpha ketoglutarate (alpha-KG), a cofactor produced by aerobic metabolism in the citric acid cycle. TET, Jumonji-family histone demethylases, and prolyl hydroxylase, a repressor of HIF1alpha under high oxygen conditions, all require alpha ketoglutarate (alpha-KG) as cofactors for their activation. The impact of the HIF1alpha and TET proteins, which appear to have opposing functions, reaches several aspects of human life-including cell growth regulation, embryonic stem cell maintenance, cell differentiation, and tumorigenesis. The role of metabolism on regulating global DNA methylation and chromatin organization is recently demanding greater attention from the biomedical research community. This article will discuss the possible role of TET activation and the regulation of 5-hmC and 5-mC levels in response to environmental stress. We will also discuss how 5-hmC and 5-mC levels at the promoters of specific genes might be a useful biomarker for exposure to environmental toxins. PMID- 21617368 TI - The influence of aging, environmental exposures and local sequence features on the variation of DNA methylation in blood. AB - In order to properly comprehend the epigenetic dysregulation that occurs during the course of disease, there is a need to characterize the epigenetic variability in healthy individuals that arises in response to aging and exposures, and to understand such variation within the biological context of the DNA sequence. We analyzed the methylation of 26,486 autosomal CpG loci in blood from 205 healthy subjects, using three complementary approaches to assess the association between methylation, age or exposures, and local sequence features, such as CpG island status, repeat sequences, location within a polycomb target gene or proximity to a transcription factor binding site. We clustered CpGs (1) using unsupervised recursively partitioned mixture modeling (RPMM) and (2) bioinformatically informed methods, and (3) also employed a marginal model-based (non-clustering) approach. We observed associations between age and methylation and hair dye use and methylation, where the direction and magnitude was contingent on the local sequence features of the CpGs. Our results demonstrate that CpGs are differentially methylated dependent upon the genomic features of the sequence in which they are embedded, and that CpG methylation is associated with age and hair dye use in a CpG context-dependent manner in healthy individuals. PMID- 21617370 TI - Sex differences in epigenetic mechanisms may underlie risk and resilience for mental health disorders. AB - Alterations in the epigenetic programming of sex differences in the brain may underlie sexually dimorphic neurodevelopmental disorders. Sex differences have been found in DNA methyltransferases 3a, DNA methylation patterns, MeCP2, and nuclear corepressor within the developing brain. Natural variations in these epigenetic mechanisms have profound consequences on gene expression and brain function. Exogenous or endogenous perturbations during development may impact these epigenetic processes and alter the trajectory of the developing brain and confer sexually dimorphic risk and resilience for developing a neurological or mental health disorder. PMID- 21617371 TI - Phytoserotonin: a review. AB - Serotonin (5-hydroxytryptamine; SER) is one of the well-studied indoleamine neurotransmitter in vertebrates. Recently SER has also been reported in wide range of plant species. The precise function of SER at the physiological level, particularly growth regulation, flowering, xylem sap exudation, ion permeability and plant morphogenesis in plant system has not been clear. Though SER is found in different parts of plant species including leaves, stems, roots, fruits and seeds, the quantity of SER within plant tissues varies widely. SER has been recently shown as a plant hormone in view of its auxin-like activity. This brief review provide an overview of SER biosynthesis, localization, its role in plant morphogenesis and possible physiological functions in plants. This would certainly help to elucidate further the multiple roles of SER in plant morphogenesis. In the future it may form the basis for studies on involvement of SER in cellular signaling mechanisms in plants. Apart from these gaps in understanding the role of SER in ontogeny of plant physiology and ecological, adaptations have been emphasized. Thus, overall perspectives in this area of research and its possible implications have been presented. PMID- 21617372 TI - The conserved fission complex on peroxisomes and mitochondria. AB - Peroxisomes are eukaryotic organelles highly versatile and dynamic in content and abundance. Plant peroxisomes mediate various metabolic pathways, a number of which are completed sequentially in peroxisomes and other subcellular organelles, including mitochondria and chloroplasts. To understand how peroxisomal dynamics contribute to changes in plant physiology and adaptation, the multiplication pathways of peroxisomes are being dissected. Research in Arabidopsis thaliana has identified several evolutionarily conserved families of proteins in peroxisome division. These include five PEROXIN11 proteins (PEX11a to -e) that induce peroxisome elongation, and the fission machinery, which is composed of three dynamin-related proteins (DRP3A, -3B, and -5B) and DRP's membrane receptor, FISSION1 (FIS1A and -1B). While the function of PEX11 is restricted to peroxisomes, the fission factors are more promiscuous. DRP3 and FIS1 proteins are shared between peroxisomes and mitochondria, and DRP5B plays a dual role in the division of chloroplasts and peroxisomes. Analysis of the Arabidopsis genome suggests that higher plants may also contain functional homologs of the yeast Mdv1/Caf4 proteins, adaptor proteins that link DRPs to FIS1 on the membrane of both peroxisomes and mitochondria. Sharing a conserved fission machine between these metabolically linked subcellular compartments throughout evolution may have some biological significance. PMID- 21617373 TI - Enhanced disease resistance to Botrytis cinerea in myb46 Arabidopsis plants is associated to an early down-regulation of CesA genes. AB - The cell wall is a protective barrier of paramount importance for the survival of plant cells. Monitoring the integrity of cell wall allows plants to quickly activate defence pathways to minimize pathogen entry and reduce the spread of disease. Counterintuitively, however, pharmacological effects as well as genetic lesions that affect cellulose biosynthesis and content confer plants with enhanced resistance against necrotrophic fungi. This kind of pathogens target cellulose for degradation to facilitate penetration and to generate glucose units as a food source. Our results points towards the existence of a transcriptional reprogramming mechanism in genes encoding cellulose synthases (CesAs) that occurs very soon after Botrytis cinerea attack and that results in a temporarily shut down of some CesA genes. Interestingly, the observed coordinated down-regulation of CesA genes is more pronounced, and occurs earlier, in myb46 mutant plants. In the resistant myb46 plants, pathogen infection induces transient down-regulation of CesA genes that concurs with a selective transcriptional reprogramming in a set of genes encoding structural cell wall proteins and extracellular remodelling enzymes. Together with previous indications, our results favour the hypothesis that CesAs are part of a surveillance system of the cell wall integrity that senses the presence of a pathogen and transduces that signal into a rapid transcriptional reprogramming of the affected cell. PMID- 21617374 TI - Potassium flux in the pollen tubes was essential in plant sexual reproduction. AB - Potassium channels are controlling K (+) transport across plasma membrane and thus playing a central role in all aspects of osmolarity as well as numerous other functions in plants including in sexual reproduction. We have used whole cell and single-channel patch-clamp recording techniques investigated the regulation of intracellular free Ca ( 2+) -activated outward K (+) channels in Pyrus pyrifolia pollen tube protoplasts. We have also showed the channels could be inhibited by heme and activated carbon monoxide (CO). In the presence of oxygen and NADPH, hemoxygenases catalyzes heme degradation, producing biliverdin, iron and CO. Considered the oxygen concentration approaching zero in the ovary, the heme will inhibit the K (+) outward flux from the intracellular of pollen tube, increasing the pollen tubes osmolarity, inducing pollen tube burst. Here we discuss the putative role of K (+) channels in plant sexual reproduction. PMID- 21617375 TI - Phosphate starvation signaling in rice. AB - Phosphorus is one of the most essential and limiting macronutrients for plants. Phosphate (Pi) deficiency could affect crop productivity seriously in agriculture. How to cope with this problem? Unveiling the molecular mechanism behind the Pi starvation responses of plants will be helpful to solve this issue. Rice is one of the most important crops, which feeds over one-third of the people in the world. In this review, we summarize the recent progress on Pi starvation signaling in rice with the intention to provide a further insight into the molecular mechanism of Pi starvation responses in rice and to give a new research direction to design transgenic plants with high Pi efficiency. PMID- 21617376 TI - Roles of heterotrimeric G proteins in guard cell ion channel regulation. AB - Stomata are formed by pairs of surrounding guard cells and perform important roles in photosynthesis, transpiration and innate immunity of terrestrial plants. Ionic solutes in the cytosol of guard cells are important for cell turgor and volume change. Consequently, transmembrane flux of ions such as K+, Cl-, and malate2- through K+ channels and anion channels of guard cells are a direct driving force for turgor change, while the opening of calcium permeable channels can serve as a trigger of cytosolic free calcium concentration elevations or oscillations, which play second messenger roles. In plants, heterotrimeric G proteins have fewer members than in animals, but they are well investigated and found to regulate these channels and to play fundamental roles in guard cell function. This mini-review focuses on the recent understanding of G-protein regulation of ion channels on the plasma membrane of guard cells and their participation in stomatal movements. PMID- 21617377 TI - Silicon efflux transporters isolated from two pumpkin cultivars contrasting in Si uptake. AB - The accumulation of silicon (Si) differs greatly with plant species and cultivars due to different ability of the roots to take up Si. In Si accumulating plants such as rice, barley and maize, Si uptake is mediated by the influx (Lsi1) and efflux (Lsi2) transporters. Here we report isolation and functional analysis of two Si efflux transporters (CmLsi2-1 and CmLsi2-2) from two pumpkin (Cucurbita moschata Duch.) cultivars contrasting in Si uptake. These cultivars are used for rootstocks of bloom and bloomless cucumber, respectively. Different from mutations in the Si influx transporter CmLsi1, there was no difference in the sequence of either CmLsi2 between two cultivars. Both CmLsi2-1 and CmLsi2-2 showed an efflux transport activity for Si and they were expressed in both the roots and shoots. These results confirm our previous finding that mutation in CmLsi1, but not in CmLsi2-1 and CmLsi2-2 are responsible for bloomless phenotype resulting from low Si uptake. PMID- 21617378 TI - Visualization of wounding-induced root-to-shoot communication in Arabidopsis. AB - It is known that wounding systemically activates the expression of various defense-related genes in plants. However, most studies of wound-induced systemic response are concerned with a leaf-to-leaf response. We have recently reported that the long distance signaling was also observed in the shoots of Arabidopsis seedling with wounded roots. We identified early and late root-to-shoot responsive (RtS) genes that were upregulated in the shoots of root-wounded seedlings at 30 min and 6 h post-injury, respectively. It is likely that the primary signals were rapidly transfered from injured roots to shoots, and then these signals were converted into chemical signals. In fact, increase of JA and OPDA content activated the expression of early and late RtS genes in shoots, respectively. In addition, we visualized wound-induced root-to-shoot response by using RtS promoter-luciferase (Luc) transgenic plants. Analysis of the AtERF13 promoter::Luc transgenic plants clearly shows that the wound-induced root-to shoot signaling was rapidly activated via the vascular systems. PMID- 21617379 TI - ATG7 contributes to plant basal immunity towards fungal infection. AB - Autophagy has an important function in cellular homeostasis. In recent years autophagy has been implicated in plant basal immunity and assigned negative ("anti-death") and positive ("pro-death") regulatory functions in controlling cell death programs that establish sufficient immunity to microbial infection. We recently showed that Arabidopsis mutants lacking the autophagy-associated (ATG) genes ATG5, ATG10 and ATG18a are compromised in their resistance towards infection with necrotrophic fungal pathogens but display an enhanced resistance towards biotrophic bacterial invaders. Thus, the function of autophagy as either being pro-death or anti-death depends critically on the lifestyle and infection strategy of invading microbes. Here we show that ATG7 contributes to resistance to fungal pathogens. Genetic inactivation of ATG7 results in elevated susceptibility towards the necrotrophic fungal pathogen, Alternaria brassicicola, with atg7 mutants developing spreading necrosis accompanied by production of reactive oxygen intermediates. Likewise, treatment with the fungal toxin fumonisin B1 causes spreading lesion formation in the atg7 mutant. We conclude that ATG7-dependent autophagy constitutes an "anti-death" ("pro-survival") plant mechanism to control the containment of cell death and immunity to necrophic fungal infection. PMID- 21617380 TI - Novel surface targets and serum biomarkers from the ovarian cancer vasculature. AB - The molecular phenotype of tumor vasculature is different from normal vasculature, offering new opportunities for diagnosis and therapy of cancer, but the identification of tumor-restricted targets remains a challenge. We investigated 13 tumor vascular markers (TVMs) from 50 candidates identified through expression profiling of ovarian cancer vascular cells and selected to be either transmembrane or secreted, and to be either absent or expressed at low levels in normal tissues while overexpressed in tumors, based on analysis of 1,110 normal and tumor tissues from publicly available Affymetrix microarray data. Tumor-specific expression of each TVM was confirmed at the protein level in tumor tissue and/or in serum. Among the 13 TVMs, 11 were expressed on tumor vascular endothelium; the remaining 2 TVMs were expressed by tumor leukocytes. Our results demonstrate that certain transmembrane TVMs such as ADAM12 and CDCP1 are selectively expressed in tumor vasculature and represent promising targets for vascular imaging or anti-vascular therapy of epithelial ovarian cancer, while secreted or shed molecules such as TNFRSF21/DR6 can function as serum biomarkers. We have identified novel tumor-specific vasculature markers which appear promising for cancer serum diagnostics, molecular imaging and/or therapeutic targeting applications and warrant further clinical development. PMID- 21617381 TI - Cuticular wax biosynthesis as a way of inducing drought resistance. AB - Plants have evolved diverse adaptive strategies to cope with drought or water deficit conditions, such as stomatal closure, maintenance of root growth and water uptake, and biosynthesis of osmoprotectants. Accumulation of cuticular waxes also contributes to drought resistance. However, it is still unclear how cuticular wax biosynthesis is regulated in response to drought and how it is associated with plant responses to drought at the molecular level. The abscisic acid (ABA)-inducible MYB96 transcription factor plays a role in drought resistance. Notably, it also regulates cuticular wax biosynthesis by binding directly to the promoters of genes encoding fatty acid elongating enzymes, such as KCS, KCR and ECR that constitute a rate-limiting step in cuticular wax biosynthesis. In the myb96-1D mutant that constitutively express the MYB96 gene, many of genes involved in cuticular wax biosynthesis are upregulated and accordingly, cuticular wax accumulation is greatly elevated. In contrast, cuticular wax accumulation is reduced in the myb96-1 mutant, linking drought with cuticular wax biosynthesis. It is evident that the MYB96 transcription factor incorporates drought stress signals into a gene regulatory network that modulates cuticular wax biosynthesis under drought stress conditions, providing a first molecular mechanism by which cuticular wax biosynthesis contributes to drought resistance and protection from pathogenic and mechanical damages as well. PMID- 21617382 TI - Challenges to understand plant responses to wind. AB - Understanding plant response to wind is complicated as this factor entails not only mechanical stress but also affects leaf microclimate. In a recent study, we found that plant responses to mechanical stress (MS) may be different and even in the opposite direction to those of wind. MS-treated Plantago major plants produced thinner more elongated leaves while those in wind did the opposite. The latter can be associated with the drying effect of wind as is further supported by data on petiole anatomy presented here. These results indicate that plant responses to wind will depend on the extent of water stress. It should also be recognized that the responses to wind may differ between different parts of a plant and between plant species. Physiological research on wind responses should thus focus on the signal sensing and transduction of both the mechanical and drought signals associated with wind, and consider both plant size and architecture. PMID- 21617383 TI - The activation of the Arabidopsis P-ATPase 1 by the brassinosteroid receptor BRI1 is independent of threonine 948 phosphorylation. AB - The plasma membrane-spanning receptor brassinosteroid insenstive 1 (BRI1) rapidly induces plant cell wall expansion in response to brassinosteroids such as brassinolide (BL). Wall expansion is accompanied by a rapid hyperpolarisation of the plasma membrane which is recordable by measuring the fluorescence lifetime (FLT) of the green fluorescent protein (GFP) fused to BRI1. For the BL induction of hyperpolarisation and wall expansion, the activation of the plasma membrane P type H+-ATPase is necessary. Furthermore, the activation of the P-ATPase requires BRI1 kinase activity and appears to be mediated by a BL-modulated association of BRI1 with the proton pump. Here, we show that BRI1 also associates with a mutant version of the Arabidopsis P-ATPase 1 (AHA1) characterized by an exchange of a well-known regulatory threonine for a non-phosphorylatable residue in the auto inhibitory C-terminal domain. Even more important, BRI1 is still able to activate this AHA1 mutant in response to BL. This suggests a novel mechanism for the enzymatic activation of the P-ATPase by BRI1 in the plasma membrane. Furthermore, we demonstrate that the FLT of BRI1-GFP can be used as a non-invasive probe to analyse long-distance BL signaling in Arabidopsis seedlings. PMID- 21617384 TI - Identification and gene expression profiling of tumor-initiating cells isolated from human osteosarcoma cell lines in an orthotopic mouse model. AB - In the cancer stem cell model a cell hierarchy has been suggested as an explanation for intratumoral heterogeneity and tumor formation is thought to be driven by this tumor cell subpopulation. The identification of cancer stem cells in osteosarcoma (OS) and the biological processes dysregulated in this cell subpopulation, also known as tumor-initiating cells (TICs), may provide new therapeutic targets. The goal of this study, therefore, was to identify and characterize the gene expression profiles of TICs isolated from human OS cell lines. We analyzed the self-renewal capacity of OS cell lines and primary OS tumors based upon their ability to form sphere-like structures (sarcospheres) under serum-starving conditions. TICs were identify from OS cell lines using the long-term label retention dye PKH26. OS TICs and the bulk of tumor cells were isolated and used to assess their ability to initiate tumors in NOD/SCID mice. Gene expression profiles of OS TICs were obtained from fresh orthotopic tumor samples. We observed that increased sarcosphere efficiency correlated with an enhanced tumorigenic potential in OS. PKH26Hi cells were shown to constitute OS TICs based upon their capacity to form more sarcospheres, as well as to generate both primary bone tumors and lung metastases efficiently in NOD/SCID mice. Genomic profiling of OS TICs revealed that both bone development and cell migration processes were dysregulated in this tumor cell subpopulation. PKH26 labeling represents a valuable tool to identify OS TICs and gene expression analysis of this tumor cell compartment may identify potential therapeutic targets. PMID- 21617386 TI - Potential role of p62 in tumor development. AB - p62 is a ubiquitously expressed cellular protein conserved in metazoa but not in plants and fungi, and is known as one of the selective substrates for autophagy. This protein is localized at the autophagosome formation site and directly interacts with LC3, an autophagosome-localizing protein, and it is incorporated subsequently into the autophagosome and finally degraded. Impaired autophagy is accompanied by the accumulation of p62, followed by the formation of aggregates positive for p62 and ubiquitinated proteins because of the nature of both self oligomerization and the ubiquitin-binding capacity of p62. The p62-positive aggregates observed in hepatocytes of liver-specific Atg7-deficient mice are completely dispersed by additional loss of p62, suggesting the involvement of p62 in the formation of disease-related inclusion bodies. Importantly, similar aggregates known as Mallory bodies and intracellular hyaline bodies have been identified in hepatocellular carcinoma (HCC). However, the pathophysiological significance of such aggregates remains unclear. Recently, we identified the role of p62-positive aggregates in human HCC and autophagy-deficient tumors in tumor development through persistent activation of Nrf2, a transcription factor responsible for stress response. PMID- 21617385 TI - Enzymatic role for soybean ecto-apyrase in nodulation. AB - Root nodulation is regulated by a variety of mechanisms. Ecto-apyrase is an enzyme proposed to control the concentration of extracellular nucleotides. Transgenic expression of the soybean GS52 ecto-apyrase was shown to stimulate nodulation. However, mutation of the enzyme to disrupt enzymatic activity prevented this effect. Therefore, the data suggest that the enzymatic activity of the ecto-apyrase is critical for nodulation enhancement, suggesting a direct effect on extracellular nucleotide hydrolysis. In this article, we propose a hypothetical mechanism for plant ecto-apyrase function during nodulation. PMID- 21617388 TI - Correction factors applied to finger dosimetry: a theoretical assessment of appropriate values for use in handling radiopharmaceuticals. AB - United States Nuclear Regulatory Commission (USNRC) regulations limit the dose to the skin to 500 mSv per year. This is also the dose limit recommended by the International Commission on Radiological Protection (ICRP). The operational quantity recommended by ICRP for quantifying dose to the skin is the personal dose equivalent, Hp(0.07) and is identical to NRC's shallow dose equivalent, Hs, also measured at a skin depth of 7 mg cm-2. However, whereas ICRP recommends averaging the dose to the skin over an area of 1 cm regardless of the size of the exposed area of skin, USNRC requires the shallow dose equivalent to be averaged over 10 cm. To monitor dose to the skin of the hands of workers handling radioactive materials and particularly in radiopharmaceutical manufacturing facilities, which is the focus of this work, workers are frequently required to wear finger ring dosimeters. The dosimeters monitor the dose at the location of the sensitive element, but this is not the dose required to show compliance (i.e., the dose averaged over the highest exposed contiguous 10 cm of skin). Therefore, it may be necessary to apply a correction factor that enables estimation of the required skin dose from the dosimeter reading. This work explored the effects of finger ring placement and of the geometry of the radioactive materials being handled by the worker on the relationship between the dosimeter reading and the desired average dose. A mathematical model of the hand was developed for this purpose that is capable of positioning the fingers in any desired grasping configuration, thereby realistically modeling manipulation of any object. The model was then used with the radiation transport code MCNP to calculate the dose distribution on the skin of the hand when handling a variety of radioactive vials and syringes, as well as the dose to the dosimeter element. Correction factors were calculated using the results of these calculations and examined for any patterns that may be useful in establishing an appropriate correction factor for this type of work. It was determined that a correction factor of one applied to the dosimeter reading, with the dosimeter placed at the base of the middle finger, provides an adequate estimate of the required average dose during a monitoring period for most commonly encountered geometries. Different correction factors may be required for exceptional or unusual source geometries and must be considered on a case-by-case basis. PMID- 21617389 TI - Organ-specific external dose coefficients and protective apron transmission factors for historical dose reconstruction for medical personnel. AB - While radiation absorbed dose (Gy) to the skin or other organs is sometimes estimated for patients from diagnostic radiologic examinations or therapeutic procedures, rarely is occupationally-received radiation absorbed dose to individual organs/tissues estimated for medical personnel; e.g., radiologic technologists or radiologists. Generally, for medical personnel, equivalent or effective radiation doses are estimated for compliance purposes. In the very few cases when organ doses to medical personnel are reconstructed, the data is usually for the purpose of epidemiologic studies; e.g., a study of historical doses and risks to a cohort of about 110,000 radiologic technologists presently underway at the U.S. National Cancer Institute. While ICRP and ICRU have published organ-specific external dose conversion coefficients (DCCs) (i.e., absorbed dose to organs and tissues per unit air kerma and dose equivalent per unit air kerma), those factors have been published primarily for mono-energetic photons at selected energies. This presents two related problems for historical dose reconstruction, both of which are addressed here. It is necessary to derive conversion factor values for (1) continuous distributions of energy typical of diagnostic medical x-rays (bremsstrahlung radiation), and (2) energies of particular radioisotopes used in medical procedures, neither of which are presented in published tables. For derivation of DCCs for bremsstrahlung radiation, combinations of x-ray tube potentials and filtrations were derived for different time periods based on a review of relevant literature. Three peak tube potentials (70 kV, 80 kV, and 90 kV) with four different amounts of beam filtration were determined to be applicable for historic dose reconstruction. The probabilities of these machine settings were assigned to each of the four time periods (earlier than 1949, 1949-1954, 1955-1968, and after 1968). Continuous functions were fit to each set of discrete values of the ICRP/ICRU mono-energetic DCCs and the functions integrated over the air-kerma weighted photon fluence of the 12 defined x-ray spectra. The air kerma-weighted DCCs in this work were developed specifically for an irradiation geometry of anterior to posterior (AP) and for the following tissues: thyroid, breast, ovary, lens of eye, lung, colon, testes, heart, skin (anterior side only), red bone marrow (RBM), and brain. In addition, a series of functional relationships to predict DT Ka-1 values for RBM dependent on body mass index [BMI (kg m-2) = weight per height] and average photon energy were derived from a published analysis. Factors to account for attenuation of radiation by protective lead aprons were also developed. Because lead protective aprons often worn by radiology personnel not only reduce the intensity of x-ray exposure but also appreciably harden the transmitted fluence of bremsstrahlung x-rays, DCCs were separately calculated for organs possibly protected by lead aprons by considering three cases: no apron, 0.25 mm Pb apron, and 0.5 mm Pb apron. For estimation of organ doses from conducting procedures with radioisotopes, continuous functions of the reported mono-energetic values were developed, and DCCs were derived by estimation of the function at relevant energies. By considering the temporal changes in primary exposure-related parameters (e.g., energy distribution), the derived DCCs and transmission factors presented here allow for more realistic historical dose reconstructions for medical personnel when monitoring badge readings are the primary data on which estimation of an individual's organ doses are based. PMID- 21617390 TI - Reconstruction of long-lived radionuclide intakes for Techa riverside residents: strontium-90. AB - Releases of radioactive materials from the Mayak Production Association in 1949 1956 resulted in contamination of the Techa River; a nuclide of major interest was 90Sr, which downstream residents consumed with water from the river and with milk contaminated by cows' consumption of river water and contaminated pasture. Over the years, several reconstructions of dose have been performed for the approximately 30,000 persons who make up the Extended Techa River Cohort. The purpose of the study described here was to derive a revised reference-90Sr-intake function for the members of this cohort. The revision was necessary because recently discovered data have provided a more accurate description of the time course of the releases, and more is now known about the importance of the pasture grass-cow-milk pathway for the members of this cohort. The fundamental basis for the derivation of the reference-90Sr-intake function remains the same: thousands of measurements of 90Sr content in bone with a special whole-body counter, thousands of measurements of beta-activity of front teeth with a special tooth beta counter, and a variety of other measurements, including post mortem measurements of 90Sr in bone, measurements of 90Sr in cow's milk, and measurements of beta activity in human excreta. Results of the new analyses are that the major intake started in September 1950 and peaked somewhat later than originally postulated. However, the total intake for adult residents has not changed significantly. For children of some birth years, the intake and incorporation of Sr in bone tissue have changed substantially. PMID- 21617391 TI - Conservatism in effective dose calculations for accident events involving fuel reprocessing waste tanks. AB - Conservatism in the calculation of the effective dose following an airborne release from an accident involving a fuel reprocessing waste tank is examined. Within the regulatory constraints at the Hanford Site, deterministic effective dose calculations are conservative by at least an order of magnitude. Deterministic calculations should be used with caution in reaching decisions associated with required safety systems and mitigation philosophy related to the accidental release of airborne radioactive material to the environment. PMID- 21617392 TI - Evaluation of the shadow shield technique for the measurement of scattered radiation. AB - ISO standard 4037 specifies the characteristics and production methods of x-ray and gamma-ray reference radiation for calibrating protection-level dosimeters and rate dosimeters. The standard limits scattered radiation to 5%. The shadow shield technique is a widely accepted test used to determine scatter contribution. Using an MCNP model, an analysis of the shadow shield technique was performed. It was found that the shadow shield technique is accurate in predicting the scatter due to the walls and floor for source-to-detector distances (R) less than 6 m. For R greater than 6 m for the modeled irradiator and calibration range, the shadow shield blocks photons that scatter upbeam from the detector, which causes the shadow shield method to under-predict the scatter percentage. Moreover, the shadow shield blocks scatter that originates from the irradiator, which causes the shadow shield method to under-predict the scatter percentage by as much as 10 percentage units for the irradiator and calibration range that were modeled. PMID- 21617393 TI - Characterization of the radiation-damaged precursor cells in bone marrow based on modeling of the peripheral blood granulocytes response. AB - Bone marrow failure is the major cause of radiation lethality in mammals. Since bone marrow is distributed heterogeneously within trabecular spongiosa encased in a cortex of cortical bone, it is very difficult to measure the extent of the radiation damage directly. However, indirect consequences of damage to marrow, such as reductions in peripheral blood cell counts, are easily measured. In this paper, the authors investgate a mathematical model of the granulopoiesis system that provides quantitative relationships between reductions in peripheral blood cells and the bone marrow precursor cells following radiation exposure. A coarse grained architecture of cellular replication and production as well as a mechanism for implicit regulation used in this model are discussed. The model is based on previous investigations of rodents. The authors test how well the model matches, in the principal dynamic regime of hematopoiesis, experimental data on large animals as well as empirical data on humans following radiation exposure. Due to its ability to infer, albeit indirectly, radiation damage to bone marrow, this model will provide a useful computational tool in radiation accident management, military operations involving nuclear warfare, radiation therapy, and space radiation risk assessment. PMID- 21617394 TI - The deviation of liver dose in real patients for thoracic computed tomography scans: a new approach to individual dosimetry with methods of radiotherapy treatment planning. AB - The increasing use of computed tomography (CT) in diagnostic imaging is associated with a relevant increase in patient dose and requires CT dose optimization. Anthropomorphic phantoms and mathematical patient models have been developed to improve the dosimetry in diagnostic imaging. Nevertheless, the doses calculated in these models and the ones individual patients can receive may differ considerably. In particular, the assessment of organ doses is problematic when organs and tissues receive only a partial exposure. A typical example for this situation is the exposure of the liver within a thoracic CT. To evaluate the impact of the field boundary and the liver volume on the individual organ dose, 50 CT scans from 25 male and 25 female patients between the ages of 27 to 87 were analyzed in this study with the volumetric tools of a treatment planning system for radiotherapy. The relative volume of the liver within a thoracic CT was assessed and compared to results from dosimetry methods using standardized patient models. The differences between an individual dose and the results from standardized patients are considerable. The fraction of the liver volume within a thoracic CT with a standard lower boundary extends from 48-92%, resulting in a possible dose difference of up to a factor of 1.7. Results from mathematical phantoms can underestimate the liver dose by more than a factor of 2.6. From the determined data, correction factors for the dosimetry of the liver using standard programs can be derived. PMID- 21617395 TI - Toward improved ionizing radiation safety standards. AB - Ionizing radiation safety standards developed by the International Commission on Radiological Protection (ICRP) during the past 50-plus years have provided guidance for effective protection of workers and the public from the potentially harmful effects of exposure to ionizing radiation, including cancer. Earlier standards were based primarily on radiation dose rate to organs of the body. More recent recommendations have calculated cancer risk as a function of cumulative dose using a linear no-threshold cancer risk model based on the acute high dose rate exposures received by the Japanese atomic bomb survivors. The underlying assumption in these current recommendations is that risk of radiation-induced cancer is proportional to cumulative dose without threshold. In conflict with this position are the studies of protracted exposures from internally-deposited radionuclides in people and laboratory animals that have demonstrated that cancer induction risk is a function of average dose rate for protracted exposures to ionizing radiation. At lower average dose rates, cancer latency can exceed natural lifespan leading to a virtual threshold. This forum statement proposes that the conflict of these two cancer risk models is explained by the fact that the increased risk of cancer observed in the atomic bomb survivor studies was primarily the result of acute high dose rate promotion of ongoing biological processes that lead to cancer rather than cancer induction. In addition, ionizing radiation-induced cancer is not the result of a simple stochastic event in a single living cell but rather a complex deterministic systemic effect in living tissues. It is recommended that the ICRP consider revising its position in light of this important distinction between cancer promotion and cancer induction. PMID- 21617398 TI - The power of partnerships: working together to promote physical activity and prevent obesity in North Carolina. PMID- 21617399 TI - Leveraging community concerns regarding meningococcal disease to address racial and socioeconomic disparities in immunization coverage. AB - Following the death of 2 adolescents due to serogroup C invasive meningococcal disease (SCIMD) in April 2008, the Chicago Department of Public Health (CDPH) observed a high level of concern in 2 Chicago communities inferred to have low meningococcal vaccine (MCV) coverage rates. In response, CDPH promptly mobilized additional resources, administering 5,343 doses of MCV in 40 schools over 2 weeks and immunizing 44% of enrolled students aged 11 to 18 years. The number of eligible students vaccinated per school ranged from 9 to 466 (median, 112) and the proportion of age-eligible students receiving the vaccine ranged from 5% to 87% (median, 52%). The attributes of the SCIMD activity did not meet the definition of a community-based outbreak, but presented an opportunity to promptly intensify existing mechanisms for meningococcal vaccination of adolescents in the affected neighborhoods and overcome traditional barriers to vaccination. PMID- 21617400 TI - Unmet needs among people reported with hepatitis C, New York City. AB - OBJECTIVE: This project sought to describe unmet needs among patients reported with hepatitis C in New York City. DESIGN: From the New York City Health Department's hepatitis C surveillance database, we randomly selected patients whose positive hepatitis C test was in April or May 2005. In 2006, we interviewed patients by telephone and collected information from their clinicians or by medical record review. SETTING: New York City. PARTICIPANTS: We interviewed 180 of the 387 eligible patients and collected information from clinicians for 145 of the 180 patients. MAIN OUTCOME MEASURES: These included whether patients had understood their clinicians' explanation of their hepatitis C diagnosis, if they had been counseled about not drinking alcohol, information about support group attendance, vaccination against hepatitis A and B, health status, treatment, and other factors. RESULTS: Of the 180 patients, 7% stated that they had not understood their clinicians' explanation of their hepatitis C diagnosis, and 26% said that they had not been counseled about avoiding alcohol. Among the 90% of patients who had not attended a hepatitis support group, 31% were interested in attending. Among the 145 patients with information from clinicians, at least 28% were susceptible to hepatitis A and 18% to hepatitis B. CONCLUSIONS: This hepatitis C surveillance project, with information from patients and clinicians, illustrates a valuable use of a chronic hepatitis C surveillance system. The patients described here had several unmet needs, including hepatitis A and B vaccination, basic information about the virus, support groups, and counseling about preventing further liver damage and preventing transmission to others. Relatively simple and affordable health department activities can address these needs, improving quality of life and decreasing the likelihood of liver disease progression. PMID- 21617401 TI - Tsunamis slosh public health practice and scholarship together: an opportunity for a healthier future? PMID- 21617402 TI - Improving collaboration-communities, health departments, and schools and programs in public health: an academic perspective. PMID- 21617403 TI - Public health, medicine, and dentistry as partners in community health: a pioneering initiative in interprofessional, practice-based education. AB - CONTEXT: As public health challenges grow more complex, the call for professional education to be interprofessional, collaborative, and grounded in real world practice has intensified. OBJECTIVE: In this article, we describe the development, implementation, and results of one pioneering course at Boston University that aims to prepare public health, medical, and dental students for their combined roles in community health settings. SETTING AND PARTICIPANTS: The Schools of Public Health, Medicine, and Dental Medicine jointly offered the course in partnership with 3 community organizations. Participants include MPH, MD, and DMD candidates. INTERVENTION: The course design integrates the use of "The Challenge Model" (created by Management Sciences for Health) with training in public health consultation techniques (eg, community-based participatory research, logic models, monitoring and evaluation). Teams of 6 to 8 medical and public health students collaborate with managers and staff of a community health center to address 1 organizational challenge and recommend a sustainability plan. RESULTS: Postcourse evaluations revealed that a cross-disciplinary, practice based education model is feasible and can meet students' learning objectives and exceed expectations of community partners. We overcame formidable obstacles related to the "silo'ed" nature of academic institutions and the competing priorities within overburdened community organizations. We found that sustained project implementation was attained at some but not all sites, yet all sites highly valued the perspective and contribution of student teams. CONCLUSION: Dynamic and replicable, this practice-based education model is adaptable to professional schools whose work intersects in the real world and calls for collaborative leadership. PMID- 21617404 TI - Health and illness in context: a pragmatic, interdisciplinary approach to teaching and learning applied public health within an urban safety net system. AB - BACKGROUND: Academic centers and community programs are too often separated by institutional and cultural chasms. Such divides weaken our capacity to develop a diverse public health-oriented, community-based workforce. This article describes one bridge designed to connect the academy to local safety net systems and the lessons learned during its construction. PROGRAM DESCRIPTION: "Health & Illness in Context" is an interdisciplinary program developed in 2008 by students at Oregon Health & Science University and staff at Portland's Central City Concern. Over a 7-week period, small cohorts of medical, nursing, and public health students gain an intimate, street-level understanding of the local safety net and the structural forces that shape it. Guided by program faculty, they traverse the maze of urban social services-following clients' pathways from homelessness and addiction to treatment, recovery, and social reintegration. In each 4-hour session, students: (1) apply key concepts from public health to challenging real world contexts, (2) explore effective, innovative approaches to addressing complex health and social issues, and (3) directly engage members of underserved communities and the diverse professionals that serve them. OUTCOMES: Although too early to formally assess its impact on career choice, Health & Illness in Context is already serving as an incubator for novel public health-oriented experiences, curricula, and activism that are further narrowing the community-university divide. Citing Health & Illness in Context as a primary inspiration, students have developed complementary elective courses, community-outreach activities, and long-term community collaborations. Meanwhile, program faculty members, now formally advise student initiatives, serve as mentors/preceptors, and have expanded their involvement at the university. PMID- 21617405 TI - NYC Epi Scholars program: promoting applied health disparities research in an urban public health department-a program model. AB - OBJECTIVE: Although health disparities research has already contributed to decreased mortality and morbidity in underserved communities, more work is needed. The NYC Epi Scholars program of the New York City Department of Health and Mental Hygiene (NYC DOHMH) aims to address gaps in critical public health needs and to train future public health leaders in epidemiology. The program is designed to increase racial/ethnic and socioeconomic diversity in the public health workforce, to provide fieldwork and practica opportunities, and to cultivate future leaders in epidemiology and public health. METHODS: Since its inception in 2007, the NYC Epi Scholars program of the NYC DOHMH has sought talented epidemiology students interested in gaining practical experience in applied health disparities research. NYC Epi Scholars is open to graduate epidemiology students who have demonstrated achievement and leadership potential and gives them an opportunity to provide high-quality research assistance to projects that identify and address health disparities of public health significance. RESULTS: Many of the program's 32 alumni have made notable contributions to public health: publishing articles in peer-reviewed journals; making presentations at national and international conferences; and after graduating, pursuing careers at the DOHMH, Centers for Disease Control and Prevention, the Environmental Protection Agency, and the National Institutes of Health. CONCLUSIONS: Because of its noted success, the NYC Epi Scholars program may serve as a "best-practice" model for expansion in other urban health departments. PMID- 21617406 TI - Community as classroom: teaching and learning public health in rural Appalachia. AB - Traditional models for public health professional education tend to be didactic, with brief, discrete practica appended. National reports of both practitioners and academicians have called for more competency-driven, interdisciplinary focused, community-based, service-oriented, and experientially-guided learning for students across the curriculum. East Tennessee State University began its own curricular revisioning in health professions education nearly 2 decades ago with a grant from the W.K. Kellogg Foundation, emphasizing competencies development through community-based learning in community-academic partnerships. This article describes 3 examples that grew from that initiative. In the first example, students in multiple classes delivered a longitudinal community-based employee wellness intervention for a rural county school district. BS public health students conducted needs assessments and prepared health education materials; MPH students conducted health assessments and worked with school wellness councils to deliver client-centered interventions; DrPH students supervised the project and provided feedback to the schools using participatory methods. In the second example, MPH students in a social-behavioral foundations course used experiential learning to investigate the region's elevated cancer mortality ranking. Following meetings with multiple community groups, students employed theoretical constructs to frame regional beliefs about cancer and presented findings to community leaders. One outcome was a 5-year community-based participatory research study of cancer in rural Appalachia. In the third example, MPH students in a health consulting course assessed local African Americans' awareness of the university's health and education programs and perceptions of their community health issues. Students learned consultation methods by assisting at multiple regional African American community meetings to discover issues and interest that resulted in the organization of a regional African American health coalition, multiple community health interventions, and the region's first health disparities summit. Lessons learned are presented which identify key elements of success and factors that influence adoption of community-based teaching and learning in public health. PMID- 21617407 TI - Integrating service-learning into an MPH curriculum for future public health practitioners: strengthening community-campus partnerships. AB - Through a 3-year grant from the Community-Campus Partnerships for Health, the Virginia Commonwealth University MPH program adopted an incremental approach to implement service-learning focused on health disparities into its curriculum. We first incorporated service-learning into an elective course and then a required internship. We then worked with the Virginia Department of Health to develop a plan for first-year students to engage in additional experiential learning through a practicum. Students also were encouraged to organize community service events, such as health fairs. Service-learning was fully incorporated into the internship. The first-year student practicum, followed by the internship, has strengthened collaborations among faculty, students, and the Virginia Department of Health and expanded student service in the community. The number of student supported community service events more than doubled. An incremental approach to incorporating service-learning led to successful implementation of the pedagogy. Service-learning benefits community partners, enriches student learning, and is well-suited for studies in public health. PMID- 21617408 TI - Infrastructure for teaching and learning in the community: Johns Hopkins University Student Outreach Resource Center (SOURCE). AB - As health professional schools strive to offer students meaningful, structured community engagement activities, various support structures are needed. In 2005, Johns Hopkins University's Schools of Medicine, Nursing, and Public Health launched the interdisciplinary community service and service-learning center, Student Outreach Resource Center (SOURCE), which operates through reciprocal partnerships between the Hopkins schools and local community-based organizations. SOURCE is recognized on campus and in the Baltimore community for its ability to recruit and prepare students to collaborate with local partners on a wide range of practice initiatives, through both curricular and cocurricular offerings. This article describes SOURCE's history and formation, process for creating authentic partnerships, services and programs, governance, and lessons learned. In a short period of time and with a modest financial investment, the expertise and infrastructure provided by SOURCE have greatly benefited both the participating community-based organizations and the Johns Hopkins health professional schools. PMID- 21617409 TI - Master of public health community-based practicum: students' and preceptors' experiences. AB - CONTEXT: We studied students' perceptions regarding a required 120-hour practicum within a program including service learning and Problem-based Learning. Responses to pre- and postpracticum surveys and a second-year survey were analyzed for 2008 to 2010 graduates. Preceptors' responses regarding students' performance were also analyzed. OBJECTIVE: To assess attainment of public health competencies by analyzing student and preceptor identification of areas for improvement. DESIGN: Basis of study was a 29-item survey regarding competencies before/after practicum completion, and a 27-item postsurvey assessing perceptions of practicum, preceptor, and site. Instruments included a 5-point Likert Scale (1 = strongly disagree/highly ineffective to 5 = strongly agree/highly effective). SETTING: Practicum experiences are selected from more than 250 community-based partners including government, health systems & not-for-profit agencies. PARTICIPANTS: Three first-year student cohorts and preceptors surveyed. Response rates varied across cohorts and instruments. MAIN OUTCOME MEASURE(S): Study focused on satisfaction scores and pre- and postsurvey response differences reported by respondents. RESULTS: Students reported high degrees of satisfaction & value in learning by doing, increased ability addressing real world problems, and commitment to working in the community. Preceptors reported satisfaction with students' competency and ability completing projects. CONCLUSION: Experiential learning in competency-based graduate public health education derived from Problem-based Learning and service learning shares commonalities and relevance with public health practice, illustrating how Problem-based Learning enhances students' ability becoming self-directed, collaborative, problem-solvers working with communities. Students' service learning ethos is manifested in value attributed to the practicum and community advocacy. Self-assessment of personal and professional development appears enhanced through the practicum experience. PMID- 21617410 TI - Evaluating community-based public health leadership training. AB - CONTEXT: Addressing the nation's increasingly complex public health challenges will require more effective multisector collaboration and stronger public health leadership. In 2005, the Healthy Wisconsin Leadership Institute launched an annual, year-long intensive "community teams" program. The goal of this program is to develop collaborative leadership and public health skills among Wisconsin based multisectoral teams mobilizing their communities to improve public health. OBJECTIVE: To measure the scope of participation and program impacts on individual learning and practice, including application of new knowledge and collective achievements of teams on coalition and short-term community outcomes. DESIGN: End-of-year participant program evaluations and follow-up telephone interviews with participants 20 months after program completion. SETTING: Community-based public health leadership training program. PARTICIPANTS: Sixty eight participants in the Community Teams Program during the years 2006 to 2007 and 2007 to 2008. MAIN OUTCOME MEASURES: Professional diversity of program participants; individual learning and practice, including application of new knowledge; and collective achievements of teams, including coalition and short term community outcomes. RESULTS: Participants in the Community Teams Program represent a diversity of sectors, including nonprofit, governmental, academic, business, and local public health. Participation increased knowledge across all public health and leadership competency areas covered in the program. Participating teams reported outcomes, including increased engagement of community leadership, expansion of preventive services, increased media coverage, strengthened community coalitions, and increased grant funding. CONCLUSIONS: Evaluation of this community-based approach to public health leadership training has shown it to be a promising model for building collaborative and public health leadership skills and initiating sustained community change for health improvement. PMID- 21617411 TI - Building sustainable community partnerships into the structure of new academic public health schools and programs. AB - We describe and assess how the College of Public Health at the University of Georgia, established in 2005, has developed formal institutional mechanisms to facilitate community-university partnerships that serve the needs of communities and the university. The College developed these partnerships as part of its founding; therefore, the University of Georgia model may serve as an important model for other new public health programs. One important lesson is the need to develop financial and organizational mechanisms that ensure stability over time. Equally important is attention to how community needs can be addressed by faculty and students in academically appropriate ways. The integration of these 2 lessons ensures that the academic mission is fulfilled at the same time that community needs are addressed. Together, these lessons suggest that multiple formal strategies are warranted in the development of academically appropriate and sustainable university-community partnerships. PMID- 21617412 TI - Professional development using student-led, community-based activities. AB - As a community health education center affiliated with an academic institution, we recognize that by investing in the professional development of our students, we not only maximize our own outcomes but those of our students as well. Our project, Creating Community Connections, was developed to aid the work of our Center in characterizing the evolving community landscape following Hurricane Katrina while providing opportunities for students to engage in experiential learning. Students in the project could gain skills in program planning and community assessment, as well as leadership and communications. Twenty-three students worked on the project during its 2 years, developing data collection tools, organizing and conducting key informant interviews, facilitating focus groups and community forums, managing data, and summarizing project findings for community presentations. Participation in this project allowed our students to grow as public health leaders and researchers while gaining a greater appreciation for community collaboration. PMID- 21617413 TI - A university, community coalition, and town partnership to promote walking. AB - Less than half of all US adults report meeting physical activity recommendations of 30 minutes or more of moderate to vigorous physical activity on at least 5 days per week. Thus, community-wide ecological initiatives are needed to create environments that support incorporating physical activity into residents' daily lives. In this article we describe an ongoing collaborative service-learning partnership between Clemson University, a community coalition, and a neighboring small rural town to address local social and physical environment supports for walking. Years 1 to 3 of this collaborative initiative were evaluated using a mixed-method approach to assess physical environment changes, social environment changes, community perceptions of support for walking, community perceptions of collaborating with university students, and students' skill development. Results revealed several key environmental changes such as mapping and marking 3 walking trails in the community, development of broad marketing efforts linked to the trails that promote community health and heritage, and annual community events to promote walking and the newly developed walking trails. Interview data with community leaders identified several key themes critical to facilitating and enhancing our university and community collaboration. Lastly, students developed skills in developing partnerships, mapping, advocacy, event planning, critical reflection, and qualitative and quantitative data collection and analysis. Through this process community members and students learn evidence-based public health skills for using data and planning frameworks to guide local initiatives, engage community members in decision making, and conducting evaluations. PMID- 21617414 TI - Reaching the underserved through community-based participatory research and service learning: description and evaluation of a unique medical student training program. AB - OBJECTIVES: To provide an overview of the Community Health Fellowship Program (CHFP), describe the types of projects completed by the community health fellows from 2005 to 2009 and to assess the program's effectiveness from the perspective of fellows and community partners. METHODS: We developed the CHFP for training medical students in community-based participatory research (CBPR), and understanding the components of successful community partnerships for addressing health disparities in underserved communities. The program has didactic and applied community research components. RESULTS: From 2005 to 2009, fellows completed 25 research projects with 19 different community partners. Fellows reported favorable attitudes about the program, their mentors, and their community projects; their research knowledge increased significantly in most areas, especially their ability to develop a succinct research question, familiarity with CBPR, and delivering a formal research presentation (Wilcoxon signed-rank test, P <.05). Community partners reported favorable attitudes toward the fellows and the program; using a 5-point Likert scale (1 = not favorable, 5 = very favorable), they reported highly favorable attitudes about fellows' level of responsibility (4.85), level of cooperation (4.85), familiarity with the needs of the medically underserved (4.69), and knowledge of how to apply local solutions to health problems (4.54). CONCLUSIONS: The CHFP has high favorability and support among fellows and community partners; the program can serve as a prototype for training future physicians in understanding and addressing the needs of the underserved, through community partnerships, and community-based participatory research. PMID- 21617415 TI - Developing leadership and advocacy skills in medical students through service learning. AB - INTRODUCTION: Traditional medical training focuses on ameliorating disease states but not on the underlying socially determined causes. The LEADS (Leadership Education Advocacy Development Scholarship) program at the University of Colorado Denver School of Medicine was designed to train medical students to become effective advocates and to promote health at the community level. METHODS: Participants in the LEADS Track complete courses in advocacy skills, perform a summer internship, and complete a mentored scholarly activity addressing population health. Students are paired with a faculty mentor and a community based organization. RESULTS: Students report empowerment, improved self-efficacy, and increased likelihood of future engagement in leadership and health advocacy. Community sponsors also rate the experience as highly valuable. CONCLUSIONS: A curriculum in advocacy and leadership skills that includes an intensive, community-based service learning experience is effective at increasing student empowerment and disposition toward community service. PMID- 21617416 TI - Recycling Mentors: an intergenerational, service-learning program to promote recycling and environmental awareness. AB - The purpose of Recycling Mentors was to implement an intergenerational, service learning program focused on promoting recycling and environmental awareness among students enrolled in Community Health (HEA 301) and Current Issues in Gerontology (GRN 440/540) and adults older than 60 years. Recycling Mentors was conducted in New Hanover County (NHC), North Carolina, where a moderate climate and coastal location attracts many tourists, retirees, and college students. A community like NHC is a good place to implement service-learning that educates both students and older adults about the benefits of recycling to individual health and the environment. During the Fall 2009 semester, undergraduate and graduate students completed institutional review board training and then conducted the program with older adults. The education component of Recycling Mentors included a pre/post survey, brochure, and scheduled visits. Overall, Recycling Mentors was positive service-learning experience with students identifying salient outcomes such as learning about recycling and the environment and working with older adults. In addition, teaching the education component of Recycling Mentors was good practice for students who will be the future health professionals. While service-learning and environmentally themed projects are common, a program that combines the 2 like Recycling Mentors is unique and has the potential to motivate individual change while positively impacting the local community and the environment. PMID- 21617417 TI - Privilege, power, and public health programs: a student perspective on deconstructing institutional racism in community service learning. AB - The Association of Schools of Public Health has identified "diversity and culture" as 1 of 7 crosscutting competencies that public health students are expected to achieve. This competency is traditionally incorporated into the curriculum through a community service-learning (CSL) component that aims to expose students to racial/ethnic health disparities. However, this model of CSL is problematic because although students are directly engaging with community members, it does not ensure long-term sustainable changes or benefits for the host community. Moreover, academic institutions have developed significant critiques of traditional CSL models where white middle-class students engage with low-income clients and communities of color, potentially reinforcing Eurocentric power and privilege. As such, public health programs require a shift in both pedagogy and curricula that more directly addresses underlying institutional racism in health disparities. Consistent with the principles of public health, a social justice framework is imperative in teaching cultural competency and should facilitate discussion of racial injustice and privilege in the students' own lives. This brief presents an autobiographical personal narrative of my experiences with CSL as a racial/ethnic minority student in a California graduate school of public health. Although autoethnography is inherently limited, this brief highlights my observations of the limitations of the service-learning model to adequately educate students on the intersection of racism and health outcomes. In addition, the brief includes suggestions for creating inclusive curricula that critically examine issues of privilege, oppression, and power dynamics related to race/ethnicity. PMID- 21617418 TI - Collaborative research: empowering students and connecting to community. AB - Public health social work is committed to improving the health and well-being of individuals and communities. Collaborative partnerships can be a tremendous resource and valuable approach to meeting community needs. This article discusses the essential role of partnership and community learning through the case study of a student-faculty team engaged in collaborative research on homelessness in upstate New York in an effort to inform the development of a community affordable housing plan. The goals of the project were to (1) improve understanding of homelessness at the local level, (2) contribute to community planning efforts to end homelessness, and (3) enrich public health social work skills. This case study describes the various ways in which collaboration is cultivated and infused throughout the life of a project. The authors reflect on benefits and challenges of collaboration, and suggest considerations for designing collaborative research projects. This article discusses the impact faculty-student and college-community collaborative partnerships can have on expanding knowledge and enhancing community well-being. PMID- 21617419 TI - Carnegie signaling the importance of community-engaged health care teaching and learning. PMID- 21617420 TI - Responding to the economy: raising the profile of vaccines for children. PMID- 21617421 TI - What hat are you wearing? PMID- 21617422 TI - The efficacy of perilesional surgical approach for foreign body granuloma. PMID- 21617423 TI - Breast reconstruction after breast cancer. AB - LEARNING OBJECTIVES: After reading this article, the participant should be able to: 1. Describe the mental, emotional, and physical benefits of reconstruction in breast cancer patients. 2. Compare the most common techniques of reconstruction in patients and detail benefits and risks associated with each. 3. Outline different methods of reconstruction and identify the method considered best for the patient based on timing of the procedures, body type, adjuvant therapies, and other coexisting conditions. 4. Distinguish between some of the different flaps that can be considered for autologous reconstruction. SUMMARY: Breast cancer is unfortunately a common disease affecting millions of women, often at a relatively young age. Reconstruction following mastectomy offers women an opportunity to mollify some of the emotional and aesthetic effects of this devastating disease. Although varying techniques of alloplastic and autologous techniques are available, all strive to achieve the same goal: the satisfactory reformation of a breast mound that appears as natural as possible without clothing and at the very least is normal in appearance under clothing. This article summarizes the various approaches to breast reconstruction and offers a balanced view of the risks and benefits of each, all of which in the end offer the opportunity for excellent and predictable results with a high degree of patient satisfaction. PMID- 21617424 TI - Visual impairment after blepharoplasty and after depot steroid injection. PMID- 21617425 TI - Lower blepharoplasty with orbital septum reconstruction using upper blepharoplasty excision material. PMID- 21617426 TI - The question mark ear deformity and its surgical repair. PMID- 21617427 TI - Temporalis fascia for lip augmentation. PMID- 21617428 TI - Human immunodeficiency virus-related lipoatrophy of the face: where should we have to fill? PMID- 21617429 TI - Neovascularization following facial surgery: possible cause and treatment. PMID- 21617430 TI - A prospective controlled assessment of microneedling with the Dermaroller device. PMID- 21617431 TI - Staged nipple reconstruction with vascularized SurgiMend acellular dermal matrix. PMID- 21617432 TI - How long does the nipple projection last after reconstruction using the skate flap purse-string technique? PMID- 21617433 TI - Bilateral autologous reconstruction from different sites: indications and outcomes after DIEP and SGAP flaps. PMID- 21617434 TI - The use of a single set of internal mammary recipient vessels in bilateral free flap breast reconstruction. PMID- 21617435 TI - The role of pocket dissection in breast implant contracture: a single surgeon's review. PMID- 21617436 TI - Lateral intercostal artery perforator flap in combination with thoracoabdominal advancement flap for correction of contour deformities following autologous breast reconstruction. PMID- 21617437 TI - Adolescent breast reduction: indications, techniques, and outcomes. PMID- 21617438 TI - Null-type anaplastic lymphoma kinase-negative anaplastic large cell lymphoma arising in a silicone breast implant capsule. PMID- 21617439 TI - Homodigital dorsal adipofascial reverse flap: clinical applications. PMID- 21617440 TI - Upper extremity Mycobacterium chelonae cutaneous infection. PMID- 21617441 TI - Our trick to control the depth of injection of the botulinum toxin in the treatment of hyperhidrosis. PMID- 21617442 TI - Free flap skin temperature correlates to microcirculatory free flap capillary blood flow. PMID- 21617443 TI - Diastasis as a cause of recurrence in ventral herniorrhaphy with porcine acellular dermal collagen implant. PMID- 21617444 TI - Panniculectomy as a pre-bariatric surgery procedure. PMID- 21617445 TI - Extended dermal apposition: a simple technique for the closure of irradiated perineal and sacral wounds. PMID- 21617446 TI - Use of a retrograde pedicled double-barreled osteocutaneous fibula flap for reconstruction of distal tibia and soft-tissue defects. PMID- 21617447 TI - Randomized controlled trials and consolidated standards of reporting trials reporting quality: an evidence-based analysis of the American Society of Plastic Surgeons, American Burn Association, and American Society for Surgery of the Hand conference meetings. PMID- 21617448 TI - Narcissism, perfectionism, and interest in cosmetic surgery. PMID- 21617449 TI - Discussion: Anaplastic large cell lymphoma and breast implants: a systematic review. PMID- 21617450 TI - A head-to-head comparison of quality of life and aesthetic outcomes following immediate, staged-immediate, and delayed oncoplastic reduction mammaplasty. AB - BACKGROUND: Oncoplastic reduction mammaplasty offers patients breast conservation with the added benefit of functional improvement in symptoms related to macromastia. The reduction can be performed in the immediate setting with the lumpectomy, in a staged-immediate fashion after final pathology has been confirmed or in a delayed fashion after completion of both lumpectomy and radiation. This study compared quality of life and aesthetic outcomes for these different cohorts. METHODS: A retrospective review was carried out on 16 consecutive patients who had oncoplastic reduction mammaplasty by the senior author (M.Y.N.) between 2003 and 2009. Demographics, oncologic treatment and timing, and reduction techniques were recorded. Patients were asked to complete a questionnaire to assess quality of life and satisfaction. Preoperative and postoperative photographs were evaluated by 15 reviewers. RESULTS: Over a 7-year period, five patients had immediate, six had staged-immediate, and five had delayed reduction mammaplasty. Mean patient age was 52.5 years, and mean body mass index was 31.5. The average timing of reduction was 0, 49, and 734 days for the three groups. Positive margins occurred in two patients, leading to completion mastectomy. In addition, one patient in the staged-immediate group had a recurrence that led to completion mastectomy. Complications occurred in seven (44 percent) of 16 patients. Questionnaire response was 75 percent (12 of 16), showing positive scores in all groups but no statistical significance. Objective aesthetic evaluation also revealed significant improvements within groups comparing various preoperative to postoperative parameters. Importantly, aesthetic scores for the delayed group were consistently lower across all aspects but did not reach significance. CONCLUSION: Oncoplastic reduction mammaplasty can be safe and effective in carefully selected patients in the immediate, staged immediate, and delayed settings. PMID- 21617451 TI - Breast fat grafting with platelet-rich plasma: a comparative clinical study and current state of the art. AB - BACKGROUND: The role of platelet-rich plasma in enhancing fat graft take is attracting the scientific community. There is, however, a lack of clinical studies on the matter. The aim of this article is to report the authors' experience in breast fat grafting with and without platelet-rich plasma and to investigate the state of the art on adipose tissue platelet-rich plasma enrichment. METHODS: The authors retrospectively reviewed 42 women who underwent breast fat grafting between September of 2007 and September of 2009. Seventeen of these patients (40 percent) were grafted with fat (according to Coleman) enriched with platelet-rich plasma at 10 percent (group A), and 25 patients (60 percent) received only fat grafts according to Coleman (group B). All patients underwent preoperative breast ultrasound and mammography and were regularly followed up with breast ultrasound 3 months later and then at 6-month intervals. The reconstructive and aesthetic outcomes were evaluated using the following parameters: (1) clinical outcomes according to the surgeons and the patient, (2) the rate of liponecrosis at breast ultrasound, and (3) the need of further fat grafting to achieve the planned result. RESULTS: The clinical outcomes, the rate of liponecrosis at breast ultrasound, and the need for further fat grafting reveal that fat grafting plus platelet-rich plasma at 10 percent is not superior to Coleman fat grafting alone. CONCLUSIONS: In the authors' retrospective analysis, no effect of platelet-rich plasma was seen in enhancing fat graft take when compared with the Coleman fat graft. Further research and prospective clinical studies are needed to understand the role of platelet-rich plasma, if any, in fat grafting. PMID- 21617452 TI - Immediate locally advanced breast cancer and chest wall reconstruction: surgical planning and reconstruction strategies with extended V-Y latissimus dorsi myocutaneous flap. AB - BACKGROUND: Surgical resection in locally advanced breast cancer produces large defects that may not be suitable for primary closure. Immediate reconstruction is controversial and presents a complicated scenario for breast surgeons and plastic surgeons. METHODS: In this study, a different design was planned for the latissimus dorsi musculocutaneous flap with primary closure in V-Y for the correction of major lesions in the anterior chest wall. Twenty-five patients underwent immediate locally advanced breast cancer reconstruction with a V-Y latissimus dorsi musculocutaneous flap. This flap was raised from adjacent tissue located on the lateral and posterior thoracic region and presented a triangular shape whose base was the lateral aspect of the mastectomy wound. The technique was indicated in patients with large thoracic wounds. RESULTS: Mean follow-up time was 16 months. Closure was obtained in the donor and recipient sites without the use of skin grafts or other more major procedures. Complications occurred in nine patients (36 percent), including dorsal wound dehiscence in five patients and seroma in three. All cases except one were treated by a conservative approach with a good result. No total flap loss was reported. All patients achieved a satisfactory thoracic reconstruction and adequate wound care. CONCLUSIONS: The V Y latissimus dorsi musculocutaneous flap is a reliable technique for immediate locally advanced breast cancer reconstruction. The technique is advantageous because the V-Y design allows primary closure of the chest wound and donor defect. Success depends on patient selection, coordinated planning with the breast cancer surgeon, and careful intraoperative management. PMID- 21617453 TI - Abdominal donor-site outcomes for medial versus lateral deep inferior epigastric artery branch perforator harvest. AB - BACKGROUND: Damage to the intercostal nerves during deep inferior epigastric perforator (DIEP) and muscle-sparing free transverse rectus abdominis musculocutaneous (TRAM) flap harvest compromises abdominal wall integrity. Intercostal motor nerves are closely associated with the lateral branch of the deep inferior epigastric artery (DIEA); therefore, the authors hypothesized that medial branch flap donor sites would develop fewer abdominal bulges/hernias. METHODS: The authors evaluated 2043 consecutive abdomen-based free flap breast reconstructions performed at The University of Texas M. D. Anderson Cancer Center between 2000 and 2010. Of these, the authors included only DIEP or muscle-sparing free TRAM flaps in which it could be clearly determined from which branch perforators were harvested. The authors examined the relationship between patient and treatment factors and donor-site hernia and bulge. RESULTS: The authors included 501 patients with a mean follow-up of 31 months [289 medial branch flaps (47 percent) and 326 lateral branch flaps (53 percent)]. Patient demographics, reconstruction timing, DIEP versus muscle-sparing free TRAM, unilateral versus bilateral distribution, and percentage of mesh closures were similar between the branch harvest groups. Twenty-eight donor sites (4.6 percent) developed a bulge/hernia. Abdominal bulge/hernia rates were similar between the medial and lateral branch donor sites (3.5 percent and 5.5 percent, respectively) (p = 0.20). CONCLUSIONS: This is the largest study to date comparing donor-site morbidity following medial or lateral DIEA branch harvest. Choice of perforators should be based on quality, size, and orientation. Medial versus lateral row perforators should not be harvested preferentially to reduce donor-site hernia or bulge. PMID- 21617454 TI - Breast reconstruction with free tissue transfer from the abdomen in the morbidly obese. AB - BACKGROUND: There are national trends of increasing incidence of morbid obesity and autologous breast reconstruction with free tissue transfer from the abdomen. The purpose of this study was to assess the safety and efficacy of free flap breast reconstruction in the morbidly obese population. METHODS: A retrospective review was conducted on all patients who underwent transverse rectus abdominis myocutaneous, deep inferior epigastric perforator, or superficial inferior epigastric artery flap breast reconstructions between July of 2006 and October of 2008. Data from all patients with a body mass index greater than 40 were compared with those of patients with a body mass index less than 40. A p value less than 0.05 was considered significant. Significant findings were then analyzed in a post hoc fashion to examine trends with increasing body mass index. RESULTS: Four hundred four patients underwent 612 free flap breast reconstructions during the study period. Twenty-five of these patients (6 percent) had a body mass index greater than 40. The morbidly obese group had significantly higher rate of total flap loss (p = 0.02), total major postoperative complications (p = 0.05), and delayed wound healing (p = 0.006). CONCLUSIONS: Free flap breast reconstruction in the morbidly obese is associated with a higher risk of total flap loss, total major postoperative complications, and delayed abdominal wound healing. However, the overall incidence of complications is low, making free tissue transfer from the abdomen an acceptable method of breast reconstruction in this patient population. PMID- 21617455 TI - Discussion: Breast reconstruction with free tissue transfer from the abdomen in the morbidly obese. PMID- 21617456 TI - Arginine improves microcirculation in the free transverse rectus abdominis myocutaneous flap after breast reconstruction: a randomized, double-blind clinical trial. AB - BACKGROUND: Partial flap loss is caused by the incapability of the vascular pedicle to provide sufficient microvascular perfusion in distal segments of the flap in addition to the reperfusion injury that occurs in the whole flap after free tissue transfer. In experimental studies, the amino acid arginine reduces reperfusion injury and improves microvascular perfusion. The purpose of this clinical study was to explore the effect of arginine in free flap surgery. METHODS: In this randomized, double blind, placebo-controlled trial, 20 patients with unilateral breast reconstruction using the free transverse rectus abdominis myocutaneous flap were included. Patient and flap data were recorded. Patients received a continuous intravenous infusion of arginine or the control amino acid alanine for 5 days. Microcirculation was recorded in the flap in a standardized fashion using laser Doppler flowmetry (Perimed). RESULTS: Zone IV microcirculatory blood flow postoperatively was higher in the arginine group than in the alanine control group (p = 0.04). CONCLUSION: The authors' study shows beneficial effects of intravenous therapy with arginine to improve microcirculation in the free transverse rectus abdominis myocutaneous flap. PMID- 21617457 TI - Virginal mammary hypertrophy: a meta-analysis and treatment algorithm. AB - BACKGROUND: Virginal mammary hypertrophy, a rapid enlargement of one or both breasts that usually presents in the adolescent years, is a rare condition that has been reported sporadically in the medical literature. Currently, there are no patient- or disease-oriented evidence-based guidelines for the treatment of this condition. This meta-analysis examines the published case reports and presents a cumulative algorithm for the diagnosis and treatment of this uncommon condition. METHODS: A literature search was performed using PubMed, with multiple keywords. Information regarding patient age, menarchal state, mass of excision, surgical technique, number of operations, pharmacologic intervention, and recurrence was extracted from each case report and analyzed using SPSS 15.1 statistical software. RESULTS: A significant relationship was found (p < 0.01), as was an odds ratio of 7.0, for the likelihood of recurrence using a reduction mammaplasty as opposed to a mastectomy. CONCLUSIONS: Based on the evidence presented in this article, certain interventions are more effective for the treatment of virginal mammary hypertrophy. On diagnosis of virginal mammary hypertrophy, tamoxifen therapy may be considered based on previous literature, barring any medical contraindications. A subcutaneous mastectomy with complete removal of breast tissue is the procedure least likely to lead to recurrence but is more deforming. Reduction mammaplasty gives an improved aesthetic breast, but it is important to counsel the patient on the likelihood of increased recurrence. Tamoxifen therapy following surgery may decrease the recurrence rate. PMID- 21617458 TI - The use of AlloDerm in postmastectomy alloplastic breast reconstruction: part I. A systematic review. AB - BACKGROUND: Postmastectomy alloplastic breast reconstruction is a common procedure that continues to evolve. Increasingly, AlloDerm is being used in both direct-to-implant and two-stage breast reconstruction. The objective of this systematic review was to summarize the outcomes from studies describing this use of AlloDerm, and to compare outcomes to those from studies reviewing non-AlloDerm alloplastic reconstruction. METHODS: A computerized search was performed across multiple databases. Studies involving patients undergoing alloplastic breast reconstruction with AlloDerm were included. A systematic review was performed to include randomized controlled trials, comparative observational studies, noncomparative observational studies, and case series. RESULTS: A systematic review of the literature revealed 14 studies that satisfied inclusion criteria. Both acute and long-term complication rates were obtained. No objective validated outcomes were reported. Ninety-three percent of included studies were level IV evidence. Complication rates were as follows: infection, 0 to 11 percent; hematoma, 0 to 6.7 percent; seroma, 0 to 9 percent; partial flap necrosis, 0 to 25 percent; implant exposure with removal, 0 to 14 percent; implant exposure with salvage, 0 to 4 percent; capsular contracture, 0 to 8 percent; and rippling, 0 to 6 percent. No study included a cost analysis. CONCLUSIONS: Complications using AlloDerm are comparable to those of non-AlloDerm alloplastic reconstructions. AlloDerm appears to confer a low rate of capsular contracture. A formal analysis is required to determine AlloDerm's cost effectiveness in use for direct-to implant reconstructions. In addition, a randomized controlled trial comparing AlloDerm use to conventional two-stage reconstruction is currently absent from the literature. PMID- 21617459 TI - The use of AlloDerm in postmastectomy alloplastic breast reconstruction: part II. A cost analysis. AB - BACKGROUND: Increasingly, AlloDerm is being used in alloplastic breast reconstruction, and has been the subject of a recent systematic review. The authors' objective was to perform a cost analysis comparing direct-to-implant with AlloDerm reconstruction to two-stage non-AlloDerm reconstruction. METHODS: Seven clinically important health outcomes and their probabilities for both types of reconstruction were derived from the recent review. A decision analytic model from the Canadian provincial payer's perspective was constructed based on these health states. Direct medical costs were estimated from a university-based hospital, yielding expected costs for direct-to-implant reconstruction with AlloDerm and two-stage non-AlloDerm reconstruction. Sensitivity analyses were conducted. RESULTS: Baseline and expected costs were calculated for direct-to implant AlloDerm and two-stage non-AlloDerm reconstruction. Direct-to-implant reconstruction with AlloDerm was found to be less expensive in the baseline ($10,240 versus $10,584) and expected cost ($10,734 versus $11,251) using a 6 * 16-cm AlloDerm sheet. With a 6 * 12-cm sheet, expected cost falls to $9673. By increasing direct-to-implant operative time from 2 hours to 2.5 hours, expected cost rises to $11,784. If capsular contracture rate requiring revision is set at 15 percent for both procedures, expected costs are $10,926 and $11,251 for direct to-implant and two-stage procedures, respectively. If the capsular contracture rate is lowered for either procedure, this has minimal impact on expected cost. CONCLUSIONS: Although AlloDerm is expensive, it appears to be cost-effective if used for direct-to-implant breast reconstruction. The methods used here may be extrapolated to different centers incorporating local costs and complication rates. A formal randomized controlled trial, including costs, is recommended. PMID- 21617460 TI - The impact of Smad3 loss of function on TGF-beta signaling and radiation-induced capsular contracture. AB - BACKGROUND: Capsular contracture remains a major problem following prosthetic breast implantation, especially in patients undergoing irradiation. Recent studies suggest that such radiation injuries are a cascading process of cytokine activation, with transforming growth factor (TGF)-beta acting as the "master switch." Because TGF-beta signals through phosphorylation of Smad3, a plausible approach to abate TGF-beta-induced capsular contracture would be to interrupt Smad3 signaling. To test this hypothesis, capsular contracture formation in wild type and Smad3 knockout mice was compared using micro-computed tomographic and histologic examination. METHODS: On day 0, 48 mice were implanted with bilateral silicone gel implants. Postoperatively, animals were imaged using live-scan micro computed tomographic scanning. Animals in the radiation arm then received a 10-Gy directed radiation dose. On postoperative days 21, 28, 35, and 42, animals were imaged again. Histologic evaluation was performed at necropsy. RESULTS: Irradiated implants in the wild-type mice demonstrated shape and contour deformation on micro-computed tomographic scanning beginning on postoperative day 21 and progressing through day 42. Conversely, micro-computed tomographic scanning of irradiated implants in knockout mice demonstrated few changes from day 0 through day 42. Corresponding histologic specimens from wild-type mice demonstrated irregular capsules composed of disorganized collagen that became thicker from day 21 to day 42. Irradiated knockout specimen maintained thin capsules from day 21 through day 42. CONCLUSIONS: In this work, inhibiting TGF beta signaling led to a reduction in radiation-induced capsular contracture as measured by micro-computed tomographic and histologic evaluation. The results of this study suggest a promising target for the prevention of capsular contracture through the development of anti-Smad3/TGF-beta-based therapies. PMID- 21617461 TI - Polymer therapy: a novel treatment to improve fat graft viability. AB - BACKGROUND: Autologous fat grafting is currently undergoing a renaissance. However, fat grafts are limited by unpredictable survival. Poloxamers can act as tissue surfactants. These nonionic surfactants have been shown to stabilize the membranes of damaged cells and to protect against injury and apoptosis in numerous models. This study was designed to investigate the ability of poloxamers to protect harvested adipocytes and to increase fat graft survival. METHODS: Lipoaspirate was obtained from surgical patients. Samples were washed in normal saline, centrifuged at 200 g, treated with various poloxamers or poloxamer components for 30 minutes, centrifuged at 200 g, and implanted into the flanks of nude mice in 1.0-cc, 1.0-g lobules. The grafts were explanted serially for 10 days and at 6 weeks. Endpoints were weight, apoptosis, cell viability, DNA content, and histology. RESULTS: Grafts treated with poloxamers P188, F108, and F127 demonstrated increased graft survival by weight. Fat grafts treated with poloxamers L64 and P188 demonstrated improvement in cell viability, and those treated with poloxamers L64, P188, and F38 demonstrated improved histology. P188 treated grafts demonstrated a 50 percent reduction in apoptosis compared with saline-treated controls (p < 0.05) and an overall 72 percent survival by weight at 6 weeks. P188 demonstrated statistically significant improvement by weight, DNA content, histology, and cell viability (89 percent versus 33 percent). CONCLUSIONS: The authors demonstrate that poloxamers, with membrane-sealing capability, can increase graft survival. Among these poloxamers, P188 demonstrated statistically significant improvement in apoptosis, graft survival by weight, cell viability, DNA content, and histology. PMID- 21617462 TI - Long-term stability of adipose tissue generated from a vascularized pedicled fat flap inside a chamber. AB - BACKGROUND: Numerous studies demonstrate the generation and short-term survival of adipose tissue; however, long-term persistence remains elusive. This study evaluates long-term survival and transferability of de novo adipose constructs based on a ligated vascular pedicle and tissue engineering chamber combination. METHODS: Defined adipose tissue flaps were implanted into rats in either intact or perforated domed chambers. In half of the groups, the chambers were removed after 10 weeks and the constructs transferred on their vascular pedicle to a new site, where they were observed for a further 10 weeks. In the remaining groups, the tissue construct was observed for 20 weeks inside the chamber. Tissue volume was assessed using magnetic resonance imaging and histologic measures, and constructs were assessed for stability and necrosis. Sections were assessed histologically and for proliferation using Ki-67. RESULTS: At 20 weeks, volume analysis revealed an increase in adipose volume from 0.04 +/- 0.001 ml at the time of insertion into the chambers to 0.27 +/- 0.004 ml in the closed and 0.44 +/- 0.014 ml in the perforated chambers. There was an additional increase of approximately 10 to 15 percent in tissue volume in flaps that remained in chambers for 20 weeks, whereas the volume of the transferred tissue not in chambers remained unaltered. Histomorphometric assessment of the tissues documented no signs of hypertrophy, fat necrosis, or atypical changes of the newly generated tissue. CONCLUSION: This study presents a promising new method of generating significant amounts of mature, vascularized, stable, and transferable adipose tissue for permanent autologous soft-tissue replacement. PMID- 21617463 TI - Expression of HIF-1alpha in ischemia and reperfusion in human microsurgical free muscle tissue transfer. AB - BACKGROUND: The aim of this study was to analyze the expression of hypoxia inducible factor (HIF)-1alpha during ischemia and after reperfusion in muscle tissue in the context of microsurgical free muscle tissue transfer. METHODS: Ten patients with soft-tissue defects needing coverage with microsurgical free muscle flaps were included in this study. In all patients, the muscle samples were taken from free myocutaneous flaps. The first sample was taken before induction of ischemia in normoxia, another one was taken after 72 +/- 11 minutes of ischemia, and the last one was taken 77 +/- 22 minutes after reperfusion. The samples were analyzed using DNA microarray, real-time polymerase chain reaction, and immunohistochemistry. RESULTS: DNA microarray, real-time polymerase chain reaction, and immunohistochemistry did not provide evidence of differential expression of HIF-1alpha comparing ischemia and reperfusion to normoxia. However, DNA microarray showed an up-regulation of activating transcription factor-3 during ischemia and spermine N1-acetyltransferase-1 during ischemia and reperfusion. CONCLUSIONS: This study shows that ischemia and reperfusion induce alterations on the gene expression level in human muscle free flaps. Data from this study indicate that the expression of HIF-1alpha might not be affected but that other putative pathways of ischemic regulation might be of great interest. Finally, these findings correspond with the surgeon's clinical experience that the accepted times of ischemia, generally up to 90 minutes, are not sufficient to induce pathophysiologic processes, which can ultimately lead to flap loss. PMID- 21617464 TI - In vitro mineralization of human mesenchymal stem cells on three-dimensional type I collagen versus PLGA scaffolds: a comparative analysis. AB - BACKGROUND: Development of a tissue engineered bone graft requires efficient bioactivity screening of biomaterials in clinically relevant three-dimensional systems. The authors analyzed the relative osteogenic potential of two three dimensional biomaterials--type I collagen and poly(L-lactide-co-glycolide) (PLGA) -to support in vitro mineralization of human mesenchymal stem cells. METHODS: Human mesenchymal stem cells were seeded onto three-dimensional PLGA or type I collagen scaffolds; incubated in osteogenic media; and harvested at 1, 4, and 7 days. Messenger RNA expression was analyzed using quantitative real-time reverse transcriptase polymerase chain reaction for osteogenic (i.e., alkaline phosphatase, osteocalcin, bone sialoprotein, Runx2/core binding factor alpha-1) and angiogenic (i.e., vascular endothelial growth factor and interleukin-8) markers. Alkaline phosphatase enzyme activity was measured at 4 and 7 days. Mineralization was detected by alizarin red staining and micro-computed tomographic imaging at 8 and 12 weeks. Mineral composition was analyzed by solid phase nuclear magnetic resonance spectroscopy. RESULTS: Early osteogenic and angiogenic markers, and alkaline phosphatase enzyme activity, were up-regulated on PLGA versus collagen scaffolds. However, long-term mineralization endpoints favored type I collagen. By 8 weeks, human mesenchymal stem cells on collagen exhibited significantly higher mineral density by micro-computed tomographic and alizarin red staining than PLGA scaffolds. Both biomaterials deposited calcium hydroxyapatite as determined by nuclear magnetic resonance spectroscopy. CONCLUSIONS: The authors' findings suggest that despite early PLGA induction of osteogenic gene expression, long-term mineralization occurs earlier and to a greater extent on type I collagen, highlighting collagen as a potential bone tissue engineering scaffold in the human mesenchymal stem cell niche. When screening the relative osteoinductive profiles of three-dimensional bone tissue engineering scaffolds in vitro, the authors recommend including long-term endpoints of osteogenesis. PMID- 21617466 TI - Human versus non-cross-linked porcine acellular dermal matrix used for ventral hernia repair: comparison of in vivo fibrovascular remodeling and mechanical repair strength. AB - BACKGROUND: Human acellular dermal matrix (HADM) and non-cross-linked porcine acellular dermal matrix (ncl-PADM) are clinically useful for complex ventral hernia repair. Direct comparisons between the two in vivo are lacking, however. This study compared clinically relevant early outcomes with these bioprosthetic materials when used for ventral hernia repair. METHODS: Seventy-two guinea pigs underwent inlay repair of surgically created hernias with HADM (n = 37) or ncl PADM (n = 35). Repair sites were harvested at 1, 2, or 4 weeks postoperatively. Adhesions were graded and quantified. Mechanical testing and histologic and immunohistologic (factor VIII) analyses of cellular and vascular infiltration were performed. RESULTS: No infections or recurrent hernias occurred. No difference was observed in mean adhesion surface area or tenacity between groups. Mean cellular infiltration (p < 0.002, weeks 1 and 4; p < 0.006, week 2) and vascular infiltration (p < 0.0003, week 1; p < 0.0001, weeks 2 and 4) were greater in HADM. Ultimate tensile strength at the implant-musculofascia interface increased over time with both materials, but no difference was observed at 4 weeks. The mean ultimate tensile strength of explanted ncl-PADM itself was consistently greater than that of HADM. The elastic modulus (stiffness) did not differ between groups at the interface but was greater in explanted ncl-PADM (p < 0.0001, weeks 1 and 2; p < 0.02, week 4). CONCLUSIONS: Both HADM and ncl-PADM become infiltrated with host cells and blood vessels within 4 weeks and have similar musculofascia-bioprosthetic interface strength. However, HADM has greater cellular and vascular infiltration. Longer-term studies will help determine whether later differences in material strength, stiffness, and remodeling affect hernia and/or bulge incidence. PMID- 21617465 TI - The fiber diameter of synthetic bioresorbable extracellular matrix influences human fibroblast morphology and fibronectin matrix assembly. AB - BACKGROUND: The ideal scaffold material should provide immediate capacity to bear mechanical loads and also permit eventual resorption and replacement with native tissue of similar mechanical integrity. Scaffold characteristics such as fiber diameter provide environmental cues that can influence cell function and differentiation. In this study, the impact of fiber diameter of scaffolds constructed from a tyrosine-based bioresorbable polymer on cellular response was investigated. METHODS: Electrospun bioresorbable poly(desamino tyrosyl-tyrosine ethyl ester carbonate) scaffolds composed of microfibers or nanofibers were constructed and seeded with human dermal fibroblasts. The impact of fiber diameter on actin cytoskeletal morphology, focal adhesion size, fibronectin matrix assembly, and cell proliferation was evaluated using immunofluorescent microscopy and computer-assisted image analysis. RESULTS: Actin stress fibers were more easily observed in cells on microfiber scaffolds compared with those on nanofiber scaffolds. Cells on nanofiber scaffolds developed smaller focal adhesion complexes compared with those on microfiber scaffolds (p < 0.0001). The temporal patterns of fibronectin matrix assembly were affected by scaffold fiber diameter, with cells on microfiber scaffolds showing a delayed response in dense fibril formation compared with nanofiber scaffolds. Cells on nanofiber scaffolds showed higher proliferation compared with microfiber scaffolds at time points under 1 week (p < 0.01), but by 2 weeks significantly higher cell proliferation was observed on microfiber scaffolds (p < 0.01). CONCLUSIONS: The fiber diameter of bioresorbable scaffolds can significantly influence cell response and suggests that the ability of scaffolds to elicit consistent biological responses depends on factors beyond scaffold composition. Such findings have important implications for the design of clinically useful engineered constructs. PMID- 21617467 TI - A histological analysis of the Mullerectomy: redefining its mechanism in ptosis repair. AB - BACKGROUND: The goals of this study were to determine whether the surgical target amount of Muller's muscle removed is consistent with the histological amount obtained, to assess the clinical outcome of ptosis repair based on these amounts, and to evaluate the role of the levator aponeurosis in the procedure. METHODS: The Mullerectomy procedure was performed on eight eyelids of four cadavers and eight eyelids of seven patients (six unilateral, one bilateral). Histologically, the dimensions of the Muller and levator muscles were measured in the specimens removed. If present, the elastic fiber network of the levator aponeurosis was noted. Ptosis measurements were analyzed according to the target amount of resection and histologic amount obtained. RESULTS: The median percentage of Muller's muscle histologically measured compared with the target tissue resection obtained was 23.25 percent (0 to 37.5 percent, n = 8) in cadavers, compared with 31.13 percent (2.8 to 58.8 percent) in patient specimens. Levator aponeurosis was identified in all cadaver specimens (n = 8) and all patient specimens (n = 8). Elastin stain identified the elastic fiber network of the levator aponeurosis. There was no statistically significant difference between amount of resection and presence of the Muller or levator muscle in cadaver (p > 0.05) or patient specimens (p > 0.05). CONCLUSIONS: There is a consistent presence of levator aponeurosis that may play a role in ptosis repair via aponeurosis advancement and resection using a modified internal Mullerectomy approach. Using the authors' surgical technique, there is no correlation of the histologically measured amount of Muller's muscle to the postoperative marginal reflex distance-1 change in the operative eyelid. The levator aponeurosis may play a greater role in ptosis repair using an internal Mullerectomy approach than previously thought. PMID- 21617468 TI - Gelsolin amyloidosis as a cause of early aging and progressive bilateral facial paralysis. AB - BACKGROUND: Gelsolin amyloidosis, or Meretoja disease, is a dominantly inherited syndrome in which the collection of amyloid leads to early aging, bilateral progressive facial paralysis, and corneal lattice dystrophy. Characteristically, the major symptoms appear in the fifth decade of life, with brow ptosis and blepharochalasis, drooping of the facial tissues, and oral disturbances. Indications and methods, as well as the results of plastic surgical treatment, seem varied. To better understand and operate on this challenging group of patients, the authors evaluated the surgical experience of their department. METHODS: From the surgical database, all patients operated on in the authors' department for gelsolin amyloidosis from 1986 to 2004 were included. Disease severity was evaluated, together with data on the course of surgery, complications, and results. RESULTS: During 1986 through 2004, 35 gelsolin amyloidosis patients underwent surgery, totaling 67 operations and 95 procedures; 89 percent consulted a plastic surgeon for brow ptosis or blepharochalasis, 15 percent for problems in oral function, and 21 percent for ocular dryness or corneal ulcers. Total bilateral forehead palsy was noted in 63 percent, total unilateral palsy in 14 percent, and weakness of the frontal branch in 23 percent. The buccal branch was affected in 40 percent. Postoperative complications were observed in 31 percent and minor complaints in 60 percent. Two-thirds, however, seemed satisfied with the results. The majority required reoperations. CONCLUSIONS: Operative treatment of gelsolin amyloidosis is symptomatic. Due to relentless disease progression, a good functional and aesthetic result seems to require selected techniques and repeated surgery. Recognizing this underdiagnosed syndrome is essential. PMID- 21617469 TI - Transoral robotic reconstructive surgery reconstruction of a tongue base defect with a radial forearm flap. PMID- 21617470 TI - Experience of an orthoplastic limb salvage team after the Haiti earthquake: analysis of caseload and early outcomes. AB - BACKGROUND: After the devastating earthquake in Haiti on January 12, 2010, a British orthoplastic limb salvage team was mobilized. The team operated in a suburb of Port-au-Prince from January 20, 2010. This analysis gives an overview of the caseload and early outcomes. METHODS: A retrospective analysis of operative data from the log book was performed from the opening of the facility on January 20, 2010, until March 12, 2010. RESULTS: In total, 348 operations were carried out on 158 patients, at an average of 47 cases per week. Seventy-three percent of the cases were soft-tissue cases and 25 percent were bony or combined soft-tissue and bony cases. The majority of bony procedures (n = 26; 16 percent) and flap procedures (n = 16; 10 percent) took place in the early weeks (weeks 1 through 4). Combined orthoplastic cases accounted for 37 percent of cases (16 of 44) in week 2 but only 7 percent (three of 43) in week 7. General anesthetic cases accounted for 89 percent of cases (39 of 44) in week 2 but only 40 percent (17 of 43) in week 7. Only six patients (4 percent) underwent amputation, but 36 operations (10 percent) dealt with the sequelae of amputation. Sixteen patients (10 percent) suffered complications, including two amputations for failed limb salvage. CONCLUSIONS: This article reports the outcomes of a limb salvage team in the acute response after an earthquake disaster with a favorable amputation rate and highlights the potential benefit of mobilizing this type of team. Detailing the changing caseload over time will allow for more efficient planning in case of a similar future disaster. PMID- 21617471 TI - Suprascapular nerve reconstruction in obstetrical brachial plexus palsy: spinal accessory nerve transfer versus C5 root grafting. AB - BACKGROUND: The purpose of this study was to determine whether there is any difference in external rotation following reconstruction of the suprascapular nerve using nerve grafts from the proximal C5 root or nerve transfer using the spinal accessory nerve. METHODS: External rotation was assessed using the Active Movement Scale immediately before surgery and 3 years postoperatively. Patients with less than 3 years of follow-up were excluded. For patients who underwent secondary shoulder surgery before the 3-year follow-up, the Active Movement Scale score before shoulder surgery was used as the outcome. RESULTS: One-hundred-six patients underwent nerve grafting, while 71 patients underwent spinal accessory nerve transfer. The spinal accessory nerve transfer group had a greater proportion of patients with total plexus palsies, more avulsions, and an earlier age at surgery (p < 0.001). In the C5 nerve graft group, the mean Active Movement Scale score increased from 0.4 to 2.2 (p < 0.001). In the nerve transfer group, the mean score increased from 0.2 to 3.0 (p < 0.001). Preoperatively, the C5 nerve graft group had significantly better scores than the nerve transfer group (p = 0.03). Postoperatively, there was no significant difference between treatments (p = 0.1). Further statistical analysis failed to demonstrate a significant advantage of one surgical treatment over the other. CONCLUSIONS: There was no difference in external rotation after suprascapular nerve reconstruction with either nerve grafting from the proximal C5 root or spinal accessory nerve transfer. The choice of suprascapular nerve reconstruction can be selected depending on specific requirements of the individual lesion. PMID- 21617472 TI - An analysis of mandibular volume in hemifacial microsomia. AB - BACKGROUND: The mandibular deformity in hemifacial microsomia is characterized by ramus-condyle unit deficiency. The Pruzansky score classifies the proximal mandible according to aberrant condylar-unit structure. The authors sought to volumetrically evaluate the hemifacial mandible compared with controls, and to assess for Pruzansky score correlation. METHODS: This is a retrospective analysis of children with hemifacial microsomia. Demographic information was obtained, and computed tomographic data were analyzed by segmentation and volumetric calculations. Age-matched controls were compared using the t test. RESULTS: Computed tomographic scans revealed 24 hemifacial and 13 controls: 62.5 percent right, 12.5 percent left, and 25 percent bilateral; and 34 percent type I, 28 percent type IIa, 16 percent type IIb, and 22 percent type III. Type IIb/III compared with type I/IIa were 11,100 and 17,773 mm, respectively (p = 0.0029). Segmental evaluation of type IIb/III versus type I/IIa showed 3590 versus 6510 mm for the proximal segments (p = 0.0022) and 7449 versus 10,829 mm for the dental bearing segments (p = 0.0221). All hemifacial microsomia hemimandibles (types I to III) were significantly less than controls: 14,837 versus 20,418 mm (p = 0.0005). Both dentate and proximal hemifacial microsomia segments statistically decreased in volume with increasing Pruzansky score. The dentate segment of the unaffected hemifacial microsomia side was statistically less than controls. CONCLUSIONS: This study volumetrically characterized the hemifacial microsomia mandibular deformity. As expected, with increasing Pruzansky severity, hemimandibular and proximal segment volumes declined. Unexpectedly, the hemifacial dentate segment also proved significantly diminished, corresponding to the degree of proximal volume loss. PMID- 21617473 TI - The double-opposing buccal flap procedure for palatal lengthening. AB - BACKGROUND: Velopharyngeal dysfunction has been treated with either a pharyngeal flap or sphincteroplasty with varying degrees of success. Both of these entities have their own series of problems, with sleep apnea and nasal mucous flow disruptions at the forefront. The purpose of this study was to review the senior author's (R.J.M.) experience performing the double-opposing buccal flap for palatal lengthening. METHODS: All patients who were treated with double-opposing buccal flaps between October of 1994 and July of 2007 were reviewed. These patients presented with varying degrees of velopharyngeal dysfunction showing some degree of velar movement at the time of surgery. Preoperative and postoperative speech results were reviewed for comparison. RESULTS: Twenty-seven patients underwent palatal lengthening, with an average length of follow-up of 58 months. Distal flap necrosis occurred in two patients. The level of intelligibility (65.4 percent versus 95.5 percent) and resonance (moderately hypernasal versus normal resonance) improved significantly postoperatively (p < 0.0001). Only one patient required the addition of a pharyngeal flap for persistent velopharyngeal dysfunction, and there were no postoperative issues with sleep apnea. CONCLUSIONS: The double-opposing buccal flap is an effective technique for lengthening the palate, improving speech, and decreasing the risks of postoperative sleep apnea. All patients experienced a dramatic improvement in their resonance and intelligibility. This technique appears most effective in patients with intact velar movement who demonstrate a small to moderate posterior velar gap. The double-opposing buccal flap is a useful means of treating velopharyngeal dysfunction, thus serving as an adjunct when improving pharyngeal closure. PMID- 21617474 TI - Primary lymphedema: clinical features and management in 138 pediatric patients. AB - BACKGROUND: Lymphedema results from maldevelopment of the lymphatic system (primary) or injury to lymphatic vasculature (secondary). Primary lymphedema is far less common than the secondary condition. The purpose of this study was to determine the clinical features of primary lymphedema in the pediatric age group. METHODS: The authors' Vascular Anomalies Center database was reviewed for patients evaluated between 1999 and 2010 with onset of lymphedema before 21 years of age. Cause, sex, age of onset, location, and familial/syndromic association were determined. Morbidity, progression, and treatment were analyzed. RESULTS: Lymphedema was confirmed in 142 children: 138 cases (97.2 percent) were primary and four (2.8 percent) were secondary. Analysis of the primary cohort showed that 58.7 percent of the patients were female. Age of onset was infancy, 49.2 percent; childhood, 9.5 percent; or adolescence, 41.3 percent. Boys most commonly presented in infancy (68.0 percent), whereas girls usually developed swelling in adolescence (55.3 percent). Lymphedema involved an extremity (81.9 percent), genitalia (4.3 percent), or both (13.8 percent). The lower limb was most commonly affected (91.7 percent), and 52.9 percent had bilateral lower extremity disease. Eleven percent of patients had familial or syndromic lymphedema. Cellulitis occurred in 18.8 percent of children; 13.0 percent required hospitalization. The majority of patients (57.9 percent) had progression of their disease. Treatment was compression garments alone (75.4 percent) or in combination with pneumatic compression (19.6 percent); 13.0 percent had operative intervention. CONCLUSIONS: Pediatric primary lymphedema usually involves the lower extremities. Boys typically are affected at birth, and girls most often present during adolescence. Most patients do not have major morbidity, are successfully managed by compression, and do not require surgical treatment. PMID- 21617475 TI - Diagnostic accuracy of maxillofacial trauma two-dimensional and three-dimensional computed tomographic scans: comparison of oral surgeons, head and neck surgeons, plastic surgeons, and neuroradiologists. AB - BACKGROUND: The authors' objectives were to study differences in diagnostic accuracy between two- and three-dimensional computed tomographic scans and among the specialties of plastic surgery, head and neck surgery, oral surgery, and neuroradiology, since this had not previously been done. METHODS: Four groups of subspecialists completed time-proctored tests of 20 maxillofacial trauma scans with zygomatic arch, zygomatic complex, orbital, Le Fort I, II, III, mandibular and panfacial fractures from five institutions (n = 40). Accuracy of diagnosis and indication for surgery, efficiency, and preference were assessed. Comparison between two- and three-dimensional scans, between expert (experienced attending) versus novice (resident/fellow), and among the four subspecialties was performed. RESULTS: For two- and three-dimensional scans, two-dimensional was more accurate for orbital floor/medial wall (40 percent and 34 percent) and frontal sinus (26 percent for diagnostic) fractures. Two-dimensional examinations took 2.3 times longer but were preferred (85 percent). Experts and novices had similar accuracy with three-dimensional scanning, but experts were more accurate with the two dimensional scanning. Experts were 3.3 times faster with two-dimensional scanning but not with three-dimensional scanning. Accuracy of diagnosis among subspecialists was similar, except that oral surgery was less accurate with orbitozygomatic fractures (79 percent versus 90 to 92 percent); neuroradiology was less accurate with indications for surgery (65 percent versus 87 to 93 percent). CONCLUSIONS: Differences in diagnostic accuracy exist between two- and three-dimensional maxillofacial scans and between expert and novice readers but not between subspecialties. Combined modalities are preferred. PMID- 21617476 TI - Breast augmentation using preexpansion and autologous fat transplantation: a clinical radiographic study. AB - BACKGROUND: Despite the increased popularity of fat grafting of the breasts, there remain unanswered questions. There is currently no standard for technique or data regarding long-term volume maintenance with this procedure. Because of the sensitive nature of breast tissue, there is a need for radiographic evaluation, focusing on volume maintenance and on tissue viability. This study was designed to quantify the long-term volume maintenance of mature adipocyte fat grafting for breast augmentation using recipient-site preexpansion. METHODS: This is a prospective examination of 25 patients in 46 breasts treated with fat grafting for breast augmentation from 2007 to 2009. Indications included micromastia, postexplantation deformity, tuberous breast deformity, and Poland syndrome. Preexpansion using the BRAVA device was used in all patients. Fat was processed using low-g-force centrifugation. Patients had preoperative and 6-month postoperative three-dimensional volumetric imaging and/or magnetic resonance imaging to quantify breast volume. RESULTS: All women had a significant increase in breast volume (range, 60 to 200 percent) at 6 months, as determined by magnetic resonance imaging (n = 12), and all had breasts that were soft and natural in appearance and feel. Magnetic resonance imaging examinations postoperatively revealed no new oil cysts or breast masses. CONCLUSIONS: Preexpansion of the breast allows for megavolume (>300 cc) grafting with reproducible, long-lasting results that can be achieved in less than 2 hours. These data can serve as a benchmark with which to evaluate the safety and efficacy of other core technology strategies in fat grafting. The authors believe preexpansion is useful for successful megavolume fat grafting to the breast. PMID- 21617477 TI - Discussion: Breast augmentation using preexpansion and autologous fat transplantation: a clinical radiographic study. PMID- 21617478 TI - Discussion: Breast augmentation using preexpansion and autologous fat transplantation: a clinical radiographic study. PMID- 21617479 TI - The safety of rhytidectomy in the elderly. AB - BACKGROUND: The purpose of this study was to evaluate the safety of face-lift surgery in an elderly population. Specifically, is chronologic age an independent risk factor leading to a higher complication rate in the elderly patient undergoing rhytidectomy surgery? METHODS: The authors retrospectively reviewed consecutive face lifts (216 patients) performed by a single surgeon over a 3-year period. Patients were divided into two groups, younger than 65 years (148 patients) and 65 years and older (68 patients). Comorbidities, operative details, and complications were compared using statistical analysis. RESULTS: The average age was 70.0 years in the elderly group and 57.6 years in the younger group. When compared with the patients younger than 65 years, elderly patients were more likely to have a higher American Society of Anesthesiologists score and to have had a prior face lift (41.2 percent versus 17.6 percent, p < 0.001). The elderly had complication rates comparable to those of younger patients (2.9 percent versus 2.0 percent major, p = 0.65; and 5.9 percent versus 6.1 percent minor, p = 0.99). There were no deaths in either group. CONCLUSIONS: In the authors' series of carefully selected elderly patients, face-lift complication rates were not statistically different when compared with those of a younger control group. The authors' data suggest that chronologic age alone was not an independent risk factor for face-lift surgery. Further studies are needed to define whether a chronologic age limit for safe face-lift surgery beyond age 65 exists. PMID- 21617480 TI - A clinical trial in facial fat grafting: filtered and washed versus centrifuged fat. AB - BACKGROUND: Although the increasing trend is to rebuild facial soft-tissue volume with autologous fat transfer, there is no agreement concerning the best way of processing the harvested fat before reinjecting it. Among all the reported fat graft processing techniques, in the present study, the authors compared the clinical results obtained using simple filtered and washed fat with those achieved by means of pure centrifuged fat. METHODS: A prospective double-blind study was conducted on 25 healthy patients undergoing facial fat transplantation from January of 2006 to June of 2006. During the same session, half the face was injected with simple filtered and washed fat, and the other half was instead treated with centrifuged fat. Subjective and objective methods were used to evaluate the results. The subjective methods included a questionnaire, sent to all patients, accompanied by an explanatory letter. The objective method involved the evaluation of preoperative and postoperative photographs by a three-member jury. The average follow-up period was 12 months. RESULTS: The authors' experimental work demonstrates that there was no significant difference between the two fat-processing techniques. In the long term, the implanted hemifacial regions produced comparable results. CONCLUSION: The authors, following their personal experience based on the reported data, went back to using the simple filtered fat after some years of use of centrifuged fat. PMID- 21617481 TI - Division or department: a microeconomic analysis. AB - BACKGROUND: In this article, the authors present a microeconomic analysis of the effects of the administrative status on plastic surgery units within academic medical centers, comparing the departmental versus subdepartmental status. The objectives are to introduce decision-making tools of microeconomics and use them to explore the potential effects of administrative status on academic plastic surgery services. METHODS: Real financial data over a decade were used to construct total cost (TC), average total cost (ATC), and total revenue (TR) curves. From these, the authors derive the efficiency scale and express the fiscal performance by examining profitability, and the commonly used ATC curve. Mathematical modeling is then used to examine the effects of departmental versus subdepartmental status, assuming that (1) a plastic surgery unit exists in a competitive market; and (2) TR > TC for the plastic surgery unit to self-sustain in the long term. The variables considered are total clinical production (Q), gross collection rates (GCR), personnel cost, and departmental tax. RESULTS: The sustainability (Q against GCR) is a hyperbolic curve with Q * GCR = TC at break even. The TC/TR = f(TR) curve resembles the ATC curve. Sectional versus departmental status for a plastic surgery service in an academic medical center depends greatly on the shape of their TC/TR = f(TR) curve. CONCLUSIONS: With most competing clinical units within the same academic medical center having departmental status, and most competing private surgeons having no institutional "taxes," the essential requirement for academic medical center plastic surgery services is to ensure that their TC/TR = f(TR) curve is comparable to that of their competitors. PMID- 21617482 TI - Authorship and medical ghostwriting: Plastic and Reconstructive Surgery policy. AB - Authorship of biomedical articles serves to acknowledge and credit individuals who contributed substantially to the development, writing, and editing of articles. In addition, authorship indicates who is responsible for ideas and experiments, and can be used to hold people accountable if something is disproven or shown false. Types of authorship can be divided into four primary categories: ghost authorship, guest authorship, gift authorship, and legitimate authorship. Of these four categories, Plastic and Reconstructive Surgery (along with other journals, the World Association of Medical Editors, the International Committee of Medical Journal Editors, the Council of Science Editors, and the Committee on Publication Ethics) affirms that only authors who meet the criteria of "legitimate authorship" as authors should be listed on an article. This article discusses what constitutes legitimate article authorship and sets forth the Journal's policy on authorship. PMID- 21617483 TI - Breast implants and anaplastic large cell lymphoma: using science to guide our patients and plastic surgeons worldwide. PMID- 21617486 TI - An algorithm for oncologic scalp reconstruction. PMID- 21617487 TI - The Cyrano nose: different treatment approaches to management of hemangiomas of the nasal tip. PMID- 21617490 TI - Finding a favorable treatment of polyacrylamide hydrogel injection complication. PMID- 21617493 TI - Factors influencing free flap management. PMID- 21617495 TI - Refinements in postoperative free flap monitoring. PMID- 21617496 TI - Single versus double venous microvascular anastomoses. PMID- 21617497 TI - One versus two venous anastomoses in microvascular free flap surgery. PMID- 21617499 TI - Some thoughts on the posterior brachioplasty. PMID- 21617501 TI - Is adherent scar always nonpliable? PMID- 21617504 TI - Six ways problem-based learning cases can sabotage patient-centered medical education. AB - PURPOSE: Problem-based learning (PBL) cases tell a story of a medical encounter; however, the version of the story is typically very biomedical in focus. The patient and her or his experience of the situation are rarely the focus of the case despite a prevalent discourse of patient-centeredness in contemporary medical education. This report describes a qualitative study that explored the question, "How does PBL teach medical students about what matters in medicine?" METHOD: The qualitative study, culminating in 2008, involved three data collection strategies: (1) a discourse analysis of a set of PBL cases from 2005 to 2006, (2) observation of a PBL tutorial group, and (3) semistructured, in depth, open-ended interviews with medical educators and medical students. RESULTS: In this report, using data gathered from 67 PBL cases, 26 hours of observation, and 14 interviews, the author describes six specific ways in which PBL cases-if not thoughtfully conceptualized and authored-can serve to overlook social considerations, thereby undermining a patient-centered approach. These comprise the detective case, the shape-shifting patient, the voiceless PBL person, the joke name, the disembodied PBL person, and the stereotypical PBL person. CONCLUSIONS: PBL cases constitute an important component of undergraduate medical education. Thoughtful authoring of PBL cases has the potential to reinforce, rather than undermine, principles of patient-centeredness. PMID- 21617505 TI - Teachers as learners: the effect of bedside teaching on the clinical skills of clinician-teachers. AB - PURPOSE: To assess the impact on full-time faculty's own clinical skills and practices of sustained clinical skills bedside teaching with preclerkship students. METHOD: This was a longitudinal, qualitative study of faculty who provide dedicated ongoing bedside clinical skills teaching for preclerkship medical students. Interviews were conducted during 2003 to 2007 with 31 faculty of the Colleges program at University of Washington School of Medicine. Content analyses of interview transcripts were performed. RESULTS: Teachers perceived a strong positive impact of teaching on their own clinical skills. Six themes were associated with the influence of bedside teaching on teachers' skills and practices. One related to deterrents to change (e.g., reliance on tests/specialists) that narrowed teachers' practice skills prior to starting bedside teaching. Three related to expansion of the process of clinical care resulting from bedside teaching: expanded knowledge and skills, deconstructing the clinical experience (e.g., deepening, broadening, slowing one's practice), and greater self-reflection (e.g., awareness of being a role model). Two were perceived outcomes: improved clinical skills (e.g., physical examination) and more mindful practices (e.g., self-confidence, patient-centered). CONCLUSIONS: Teachers perceived profound positive impact on their clinical skills from teaching preclerkship students at the bedside. Further studies are needed, including comparing teaching preclerkship students with teaching advanced students and residents, to assess whether teaching at other levels has this effect. PMID- 21617506 TI - A history of medical student debt: observations and implications for the future of medical education. AB - Over the last 50 years, medical student debt has become a problem of national importance, and obtaining medical education in the United States has become a loan-dependent, individual investment. Although this phenomenon must be understood in the general context of U.S. higher education as well as economic and social trends in late-20th-century America, the historical problem of medical student debt requires specific attention for several reasons. First, current mechanisms for students' educational financing may not withstand debt levels above a certain ceiling which is rapidly approaching. Second, there are no standards for costs of medical school attendance, and these can vary dramatically between different schools even within a single city. Third, there is no consensus on the true cost of educating a medical student, which limits accountability to students and society for these costs. Fourth, policy efforts to improve physician workforce diversity and mitigate shortages in the primary care workforce are inhibited by rising levels of medical student indebtedness. Fortunately, the current effort to expand the U.S. physician workforce presents a unique opportunity to confront the unsustainable growth of medical student debt and explore new approaches to the financing of medical students' education. PMID- 21617507 TI - The association between intolerance of ambiguity and decline in medical students' attitudes toward the underserved. AB - PURPOSE: Little published research details the risk factors for the decline in students' attitudes toward underserved populations during medical school. The authors assessed the association between this attitude change and intolerance of ambiguity (the tendency to perceive novel or complex situations as sources of threat), since treating underserved populations often involves a high level of complexity. METHOD: The University of New Mexico School of Medicine administered a survey assessing attitudes toward underserved populations at matriculation and at graduation to seven consecutive medical school classes (matriculation years 1999 to 2005). The university also administered a survey measuring tolerance of ambiguity at matriculation. Five hundred twenty-nine students were eligible to complete both surveys between 1999 and 2009. RESULTS: Three hundred thirteen (59%) students completed the attitude survey at matriculation and graduation. Attitude scores for a majority of students (69%) decreased from matriculation to graduation. Changes in scores ranged from +25 to -35; the average change was 4.5. Linear regression analysis showed that those who were tolerant of ambiguity (top 20% of tolerance of ambiguity scores) were significantly less likely to have declines in attitudes toward the underserved; the coefficient was 3.69 (P = .003). Other factors independently associated with maintaining high attitude scores were being female and starting medical school at age 24 or younger. CONCLUSIONS: Attention to, and practice with, ambiguous situations may help moderate decreases in attitudes toward underserved populations. Medical education should address the fact that physicians face much ambiguity and should offer students tools to help them respond to ambiguous clinical situations. PMID- 21617508 TI - Perspective: call to action: it is time for academic institutions to appoint a resident quality and patient safety officer. AB - In meeting the Accreditation Council for Graduate Medical Education (ACGME) core competency requirements, teaching hospitals often find it challenging to ensure effective involvement of housestaff in the area of quality and patient safety (QPS). Because housestaff are the frontline providers of care to patients, and medical errors occasionally occur based on their actions, it is essential for health care organizations to engage them in QPS processes.In early 2008 a Housestaff Quality Council (HQC) was established at New York-Presbyterian Hospital, Weill Cornell Medical Center, to improve QPS by engaging housestaff in policy and decision-making processes and to promote greater housestaff participation in QPS initiatives. It was quickly realized that the success of the HQC was highly contingent on alignment with the institution's overall QPS agenda. To this end, the position of resident QPS officer was created to strengthen the relationship between the hospital's strategic goals and the HQC. The authors describe the success of the resident QPS officers at their institution and observe that by appointing and supporting resident QPS officers, hospitals will be better able to meet their quality and safety goals, residency programs will be able to fulfill their required ACGME core competencies, and the overall quality and safety of patient care can be improved. Simultaneously, the creation of this position will help to create a new cadre of physician leaders needed to further the goals of QPS in health care. PMID- 21617509 TI - Using behavior change plans to improve medical student self-care. AB - PURPOSE: To describe an innovative approach to teaching medical students the principles and practice of health behavior change and self-care using a behavior change plan (BCP). METHOD: Second-year medical students at Northwestern University Feinberg School of Medicine (n = 343) took a required Healthy Living unit in 2008 or 2009. They completed a BCP project in which they selected a personal behavior to change (exercise, nutrition, sleep, personal habits/hygiene, study/work habits, or mental/emotional health), set a goal, tracked progress, and self-assessed success. The authors employed a one-group posttest-only design to conduct a quantitative analysis and a qualitative evaluation of students' BCPs and their attitudes concerning the project. RESULTS: Among the 343 students, 299 (87.2%) set BCP goals related to exercise, nutrition, or sleep. BCP outcomes varied: 139 students (40.5%) achieved their goal, 170 (49.6%) failed to do so, and 34 (9.9%) were uncertain. Factor analysis produced two independent attitude scales: utility (alpha = .80) and burden (alpha = .67). Logistic regression showed that success approached statistical significance only in the sleep behavior category and for the utility attitude scale. Qualitative case reports provide insights about BCP targets, management, and results. After completing the assignment, 274 (79.9%) of the students considered themselves to be healthier, and 281 (81.9%) indicated they would use the process again. CONCLUSIONS: Completing a BCP is a valuable and effective exercise that enables medical students to practice the strategies and skills and experience the obstacles of changing health behavior. PMID- 21617510 TI - Caring for underserved patients through neighborhood health screening: outcomes of a longitudinal, interprofessional, student-run home visit program in Singapore. AB - PURPOSE: Service learning, an effective vehicle for teaching undergraduate public health while providing underserved communities with medical care, is not well established in Asia. The authors evaluated a service learning program, Neighborhood Health Screening (NHS), in Singapore. METHOD: Medical and nursing undergraduate students provided in-home medical services to patients in a low income neighborhood (January-June 2010). The authors assessed student-reported pedagogical effectiveness in nine domains, asked students for qualitative feedback on their experiences, assessed patients' satisfaction with NHS, and tracked clinical outcomes. RESULTS: Of the 240 medical and 34 nursing students who participated, 222 (93%) and 34 (100%), respectively, completed the questionnaire; 136 of the medical students (57%) also provided qualitative feedback. Most students felt NHS was beneficial across all domains. Male medical students were less likely to report increased understanding of deficiencies in the health care system and long-term management of chronic disease; preclinical students were more likely to report improvements in comprehending ethical issues, critical thinking and action skills, and gaining and applying knowledge. Qualitative feedback supported quantitative findings. Patients were satisfied with NHS: 266 (75%) agreed that NHS improved their health, and 301 (85%) felt NHS provided sufficient time to address their issues. After a single year, amongst patients with known hypertension, treatment increased from 63% to 93% (P < .001), and blood pressure control amongst those who were on treatment improved from 42% to 79% (P < .001). CONCLUSIONS: Service learning can make an important contribution to medical teaching and patient care in Asia. PMID- 21617511 TI - Contributors of black men's success in admission to and graduation from medical school. AB - PURPOSE: To determine characteristics and individual experiences that contribute to black men's success in being admitted to and graduating from medical school. METHOD: In 2010, one of the authors, a black man, interviewed 10 black male medical students enrolled at Florida State University College of Medicine and 3 black male physicians associated with that school, using consensual qualitative research methodology to analyze the data. The investigators recorded and transcribed the interviews, coded them to determine themes, and identified an overarching theoretical construct. RESULTS: The authors identified six broad contributors to successful admission to and completion of medical school: social support, education, exposure to the field of medicine, group identity, faith, and social responsibility. The six categories were subsequently grouped into four major themes: educational experiences, exposure to medicine, psychosocial cultural experiences, and personal attributes and individual perceptions. The metaphor of a table (success) with four legs (four major themes) illustrates the complex dynamics that contribute to success. CONCLUSIONS: The success of black men and the factors that contribute to that success guided this study. The results reinforced the conclusion that success for black men is achieved via a balance between educational experiences, exposure to medicine, psychosocial cultural experiences, and personal attributes and individual perceptions. This information can be used by medical schools to strengthen their outreach programs, provide a theoretical construct for discussion and research, and generate questions for future quantitative studies. PMID- 21617512 TI - Perspective: medical education research and the institutional review board: reexamining the process. AB - Medical school and residency training curricula across the country have undergone extensive revisions and, much like clinical quality improvement (QI) initiatives, require assessments of new programs. Because sharing knowledge is a hallmark of academic medicine, program evaluation may come under the purview of the institutional review board (IRB); however, the distinction between QI and research is often unclear. And yet a medical education (ME) inquiry can be designed according to either paradigm. The purpose of this article is to bring IRBs and ME researchers closer to a shared understanding of key concepts underlying human participation in research and QI activities, and to consensus on the application of these concepts. The current QI discourse provides a useful framework for making this distinction; the authors identify key theoretical principles and practical considerations derived from this work that are relevant to ME and training, such as the application of the regulatory definition of human subject research to ME inquiries. For ME inquiries defined as human subject research, and therefore subject to IRB review, this article explores the application of the human research regulations to ME research. It concludes with practical suggestions for institutions, IRBs, and ME researchers, which range from formal procedures for making the QI versus research distinction, to instruction in study design and development and the human subject regulatory implications. The intent is to promote a discussion that will result in greater consensus and a more consistent application of the regulatory framework. PMID- 21617513 TI - The key role of a transition course in preparing medical students for internship. AB - Among the core transitions in medical education is the one from medical school to residency. Despite this challenging transition, the final year of medical school is known as lacking structure and clarity. The authors examine the preparation of medical students for the professional and personal challenges of internship in the context of transition courses. They first describe the development of a residency transition course, offered since 2001 at the University of California, San Francisco, School of Medicine (UCSF), exploring aspects of a needs assessment, course goals and content, core competencies, and course implementation. They then critically analyze the course, judging it successful based on high subjective satisfaction scores and increased perceived preparedness data. Next, the authors discuss the national context of transition courses, perspectives of various stakeholders, and lessons learned from the UCSF experience. Finally, they consider future directions, suggesting that internship transition courses be a standard part of the medical school curriculum. PMID- 21617514 TI - Interprofessional education in the internal medicine clerkship: results from a national survey. AB - PURPOSE: Growing data support interprofessional teams as an important part of medical education. This study describes attitudes, barriers, and practices regarding interprofessional education (IPE) in internal medicine (IM) clerkships in the United States and Canada. METHOD: In 2009, a section on IPE was included on the Clerkship Directors in Internal Medicine annual survey. This section contained 23 multiple-choice questions exploring both core and subinternship experiences. Data were analyzed using descriptive statistics and Rasch analysis. RESULTS: Sixty-nine of 107 institutional members responded to the survey (64% response rate). Approximately 68% of responding clerkship directors believed that IPE is important to the practice of IM. However, only 57% believed that it should become a part of the undergraduate clinical curriculum. The three most significant barriers to IPE in the IM clerkship were scheduling alignment, time in the existing curriculum, and resources in time and money. Although more than half of respondents felt IPE should be included in the clinical curriculum, 81% indicated that there was no formal curriculum on IPE in their core IM clerkship, and 84% indicated that there was no formal curriculum during IM subinternship rotations at their institution. CONCLUSIONS: There is limited penetration of IPE into one of the foundational clinical training episodes for medical students in Liaison Committee for Medical Education-accredited schools. This may be related to misperceptions of the relative value of these experiences and limitations of curricular time. Learning in and from successful models of interprofessional teams in clinical practice may help overcome these barriers. PMID- 21617515 TI - An innovative approach for calculating the work relative value units of clinical activities otherwise concealed. AB - Since the introduction of work relative value units (wRVUs), academic institutions have increasingly relied on them as a means of assessing clinical productivity. However, wRVUs do not capture all activities performed in the academic setting. Although proposals exist for converting teaching, research, and administrative activities into wRVUs, certain clinical activities-deemed "special services" at the University of Kentucky-remain unrecognized by this metric. For instance, wRVUs do not capture activities which include clinical work performed on a contractual basis, nor do they capture medicolegal activities. Yet, these and other special services often represent an important stream of revenue for an academic department. Because of both the significant dependence of wRVUs in determining clinical productivity and the failure of wRVUs to capture special services, the authors propose a formula that converts these clinical efforts into wRVUs. PMID- 21617516 TI - Exploring the changing learning environment of the gross anatomy lab. AB - PURPOSE: The objective of this study was to assess the impact of virtual models and prosected specimens in the context of the gross anatomy lab. METHOD: In 2009, student volunteers from an undergraduate anatomy class were randomly assigned to study groups in one of three learning conditions. All groups studied the muscles of mastication and completed identical learning objectives during a 45-minute lab. All groups were provided with two reference atlases. Groups were distinguished by the type of primary tools they were provided: gross prosections, three-dimensional stereoscopic computer model, or both resources. The facilitator kept observational field notes. A prepost multiple-choice knowledge test was administered to evaluate students' learning. RESULTS: No significant effect of the laboratory models was demonstrated between groups on the prepost assessment of knowledge. Recurring observations included students' tendency to revert to individual memorization prior to the posttest, rotation of models to match views in the provided atlas, and dissemination of groups into smaller working units. CONCLUSIONS: The use of virtual lab resources seemed to influence the social context and learning environment of the anatomy lab. As computer-based learning methods are implemented and studied, they must be evaluated beyond their impact on knowledge gain to consider the effect technology has on students' social development. PMID- 21617517 TI - Consultancies: a model for interdisciplinary training and mentoring of junior faculty investigators. AB - The study of complex, health-related problems is often best addressed by interdisciplinary teams, and yet models for training and mentoring junior investigators in an interdisciplinary mode are not widely available. Here, the authors describe their school's version of the consultancy process, a two-year effort (September 2007 to June 2009) sponsored by the University of Washington's Center for Interdisciplinary Geriatric Research, as a model for short-and long term, interdisciplinary training and mentoring of junior faculty investigators, and evaluate its effects on establishing productive cross-disciplinary linkages among them. Between September 2007 and December 2008, written feedback was collected from participating faculty after each consultancy session. A brief, Internet-based survey of all attendees was conducted in February 2009 to gather information about longer-term implications and benefits of consultancy participation. Most respondents rated sessions highly, and a majority of the respondents reported increased networking opportunities, access to resources, new research questions, access to expertise beyond their disciplines as a result of the sessions, and a positive impact on cross-disciplinary collaborations. Their responses suggest that the consultancy format may stimulate the formation of new interdisciplinary mentoring relationships and foster cross-disciplinary collaborations. PMID- 21617518 TI - Secondary hyperparathyroidism in HIV patients: is there any responsibility of highly active antiretroviral therapy? AB - Secondary hyperparathyroidism may develop in the presence of hypovitaminosis D in order to maintain calcium homeostasis. We conducted a cross-sectional analysis in a cohort of 371 patients, identifying secondary hyperparathyroidism in 65 patients. This high prevalence (17.5%) was in part justified by the high prevalence of hypovitaminosis D (77.4%) in the whole sample, but we also identified an independent association with the use of tenofovir. PMID- 21617519 TI - Measuring concurrent partnerships: potential for underestimation in UNAIDS recommended method. AB - Research on concurrent sexual partnerships has been constrained by inconsistent and inaccurate measurements of these partnerships. Recently, a UNAIDS working group recommended a method based on sexual partner histories to improve the measurement of concurrency. Using recent survey data for young adults living in Cape Town, South Africa, we found that this method could result in underestimates of concurrency due to respondents' failure to report additional sexual partners. PMID- 21617520 TI - Apolipoprotein E polymorphism and behavioral and psychological symptoms of dementia in patients with Alzheimer disease. AB - The aims of this study were to identify subsyndromes of behavioral and psychological symptoms of dementia (BPSD) in Alzheimer disease (AD), and to investigate whether the apolipoprotein E (ApoE) gene confers a risk of distinct BPSD subsyndromes. BPSD of 96 patients with AD were assessed using the Neuropsychiatric Inventory. Factor analysis with principal component analysis and varimax rotation was used to construct the BPSD subsyndromes. ApoE genotypes were determined using the TaqMan technology. The results showed that the 5 subsyndromes can be determined, including: agitation/aggression-delusion, euphoria-disinhibition, depression-apathy, hallucination-nighttime behavior, and appetite. ApoE epsilon4 carriers had higher factor scores in the agitation/aggression-delusion subsyndrome. We demonstrated that ApoE epsilon4 confers a higher risk for the subsyndrome of agitation/aggression delusion in AD. PMID- 21617522 TI - HER2 expression in gastric and gastroesophageal junction adenocarcinoma in a US population: clinicopathologic analysis with proposed approach to HER2 assessment. AB - Recent evidence suggests that trastuzumab, a monoclonal antibody which targets HER2, in combination with chemotherapy is a therapeutic option in patients with HER2-positive gastric or gastroesophageal junction cancer. Widely accepted guidelines for HER2 testing in gastric and gastroesophageal junction cancer have not been established. The purpose of this study was to analyze the incidence and patterns of HER2 expression in gastric and gastroesophageal junction cancer using a tissue microarray approach, which closely simulates small biopsies routinely tested for HER2. One hundred sixty-nine patients, including 99 primary gastric adenocarcinomas and 70 primary gastroesophageal junction carcinomas were analyzed for HER2 overexpression by immunohistochemistry and HER2 gene amplification by fluorescence in situ hybridization using scoring schemes proposed by both American Society of Clinical Oncology/College of American Pathologists (ASCO/CAP) and the results of the recently published Trastuzumab for Gastric Cancer (ToGA) trial. In our analysis, 19 adenocarcinomas were HER2 positive, defined as either a HER2/CEP17 ratio >2.2 and/or a 3+ HER2 immunohistochemistry score with either the ASCO/CAP or ToGA scoring schemes. Of the 19 HER2-positive adenocarcinomas, 8 (42%) exhibited a characteristic strongly intense basolateral membranous staining pattern which would be interpreted as negative (1+) using the accepted ASCO/CAP scoring scheme for HER2 assessment in breast carcinoma, but were correctly labeled as 3+ positive using the proposed ToGA scoring scheme. Of the 19 HER2 positive adenocarcinomas, 8 (42%) demonstrated heterogeneous HER2 protein expression by immunohistochemistry. Twelve of 99 (12%) gastric carcinomas were positive for HER2. Of these, HER2 was more often identified in intestinal-type adenocarcinomas (10 of 52, 19%) compared with diffuse (2 of 34, 6%) adenocarcinoma. Seven of 70 (10%) gastroesophageal junction carcinomas were positive for HER2 of which all were intestinal type (7 of 58, 12%). HER2 status or primary tumor site did not correlate with patient survival. Gastric and gastroesophageal junction adenocarcinomas typically display a characteristic basolateral membranous pattern of HER2 expression which is often heterogeneous rendering routine evaluation of HER2 status on small tissue samples challenging. PMID- 21617521 TI - Prevalence of psychotic symptoms in a community-based Parkinson disease sample. AB - OBJECTIVES: : To determine the prevalence of psychotic phenomena, including minor symptoms, in a Parkinson disease (PD) sample and compare the clinical correlates associated with the various psychotic phenomena. To evaluate the extent to which cases met National Institute of Neurological Diseases and Stroke (NINDS)/National Institute of Mental Health (NIMH)-proposed criteria for PD-associated psychosis. METHODS: : A total of 250 patients with idiopathic PD and Mini Mental State Exam scores greater than 23 from three community-based movement disorder clinics underwent comprehensive research diagnostic evaluations by a geriatric psychiatrist as part of a study on mood disorders in PD. Psychotic symptoms were categorized using a checklist, which included a breakdown of hallucinations, delusions, and minor symptoms. Clinical characteristics of groups with minor and other psychotic symptoms were compared. The NINDS/NIMH criteria for PD-psychosis were retrospectively applied. RESULTS: : Of the total sample, 26% of patients were found to have any current psychotic symptoms, with 47.7% of those having isolated minor symptoms, and 52.3% having hallucinations and/or delusions. Compared to those with no current psychiatric symptoms, minor symptoms were associated with more depressive symptoms and worse quality of life, and 90.8% of those with psychotic symptoms fulfilled the NINDS/NIMH proposed criteria. CONCLUSIONS: : Psychotic symptoms are common in PD patients, with minor psychotic phenomena present in nearly half of affected patients in a community-based sample. Psychotic symptoms, including minor phenomena, were clinically significant. The NINDS/NIMH PD-psychosis criteria captured the clinical characteristics of psychosis as it relates to PD. Longitudinal studies are needed to determine whether minor psychotic symptoms represent a precursor to hallucinations and delusions, and to further validate diagnostic criteria. PMID- 21617523 TI - CD46 Expression is an unfavorable prognostic factor in breast cancer cases. AB - The membrane cofactor protein, CD46 represents a complement inhibitor, which protects autologous cells from complement-mediated cytotoxicity. On tumor cells, CD46 may exhibit the potential to protect them from immune responses of the host. The present study aimed at evaluation of prognostic significance of CD46 expression in breast cancers. The analyses were performed on 70 samples of breast cancer. Immunohistochemical reactions were performed on paraffin sections of studied tumors using monoclonal antibodies directed against CD46. Results of the immunohistochemical reactions and of clinical observations were subjected to statistical analysis. Multivariate analysis showed that expression of CD46 and involvement of lymph nodes represent independent risk factors for disease-free survival and overall survival. Kaplan-Meier analysis showed that patients with tumors negative for CD46 have an increased progression-free time and overall survival time as compared with patients with the CD46-positive tumors. The study demonstrates that breast cancers manifest CD46 expression and that it is linked to a less favorable prognosis. PMID- 21617524 TI - Distinction of high-grade neuroendocrine carcinoma/small cell carcinoma from conventional urothelial carcinoma of urinary bladder: an immunohistochemical approach. AB - CONTEXT: High-grade neuroendocrine carcinomas and small cell carcinomas (HGNEC/SmCC) of the urinary bladder are uncommon but aggressive neoplasms. Differentiation of HGNEC/SmCC from high-grade urothelial carcinoma (UC) is based on histomorphologic features, but can be difficult in small biopsies and cases with mixed morphology. OBJECTIVE: We attempt to identify a limited immunohistochemical panel that aids in this distinction. DESIGN: We selected 39 cases of bladder carcinoma with small cell morphology: 7 HGNEC/SmCC, 21 high grade UC with neuroendocrine-like pattern, and 11 mixed neoplasms. Immunohistochemistry for pan-cytokeratin, synaptophysin, chromogranin, p63, and thyroid transcription factor-1 was performed. RESULTS: Pan-cytokeratin was positive in 6 of 7 cases (86%) of the HGNEC/SmCC group. All 7 tumors were positive for synaptophysin, 6 of them were negative for p63 and chromogranin, and 1 was positive for p63 and chromogranin. All 21 high-grade UC with neuroendocrine like pattern of growth showed positive staining for pan-cytokeratin, and were all negative for synaptophysin and chromogranin. Sixteen (76%) of high-grade UC were also positive for p63. All 11 mixed tumors were positive for pan-cytokeratin. In 10 of the 11 mixed tumors (91%), synaptophysin was positive in the neuroendocrine differentiated areas and it was negative in the urothelial component. In 2 of the 11 mixed tumors (18%) chromogranin was also positive. Three (27%) of the 11 mixed cases were positive for p63 in the UC foci. Chromogranin was negative in 6 of the pure HGNEC/SmCC and in 8 of the mixed tumors. None of the 39 samples were reactive for thyroid transcription factor-1. CONCLUSIONS: A limited immunohistochemical panel including pan-cytokeratin, synaptophysin, and p63 discriminates HGNEC/SmCC from high-grade UC. PMID- 21617525 TI - Epidemiological study of injuries in men's international under-20 rugby union tournaments. AB - OBJECTIVE: To determine the incidence, nature, and causes of match injuries sustained during Under-20 (U-20) international rugby. DESIGN: Prospective cohort study; definitions and procedures were compliant with the international consensus statement for epidemiological studies in rugby. SETTING: 2008 and 2010 IRB U-20 Junior World Championships and Junior World Rugby Trophies. PARTICIPANTS: Nine hundred forty-one players representing 35 international teams. ASSESSMENT OF RISK FACTORS: Injuries reported as functions of playing position and nature and cause of injury. MAIN OUTCOME MEASURES: Incidence, location, type, severity, and causes of match injuries. RESULTS: Incidence of injury was 57.2 per 1000 player-match hours (forwards, 55.3; backs, 59.4) with a mean severity of 22.4 days (forwards, 27.7; backs, 16.9) and a median severity of 6 days (forwards, 8; backs, 6). Lower limb ligament (25.3%) and muscle (21.3%) pathologies were the main injuries. Most injuries were acute (90.4%) and sustained during tackles (45.1%) and collisions (17.7%). CONCLUSIONS: The study showed that the overall risk of injury for players in international U-20 rugby is significantly lower than that reported at the full international level of play; the nature and causes of injury at U-20 are similar to those at the full international level of play. PMID- 21617526 TI - Evaluation of standard endotracheal intubation, assisted laryngoscopy (airtraq), and laryngeal mask airway in the management of the helmeted athlete airway: a manikin study. AB - OBJECTIVES: Physicians at sporting events must rarely manage the airway of a helmeted athlete. This poses challenges for providers who do not regularly engage in airway management. In a manikin model, our purpose was to determine (1) if standard endotracheal intubation (ETI) of a simulated helmeted athlete is adversely affected by bright-light conditions and (2) if the use of laryngeal mask airway (LMA) or Airtraq improves airway management success. DESIGN: This is a randomized, prospective, crossover study. SETTING: The study was conducted at a 500-bed community-based hospital with residency training programs in family medicine and emergency medicine, as well as a fellowship in sports medicine. PARTICIPANTS: We randomized 42 residents to manage the airway of a simulated helmeted athlete in c-spine immobilization using ETI, Airtraq, and LMA. Each method was attempted under bright light and in standard light. MAIN OUTCOME MEASURES: Our main outcomes were success or failure of airway and time to airway. Secondary outcome was perceived difficulty in airway management as a factor of environmental factors. RESULTS: Airway success rates were 93% for ETI, 99% for LMA, and 75% for Airtraq. Standard ETI was significantly faster than intubation using the Airtraq (P = 0.0001) and had greater success (P = 0.004). Time to airway was faster with LMA than with standard ETI (P < 0.00001). There was no impact of bright light on ETI time (P = 0.61). CONCLUSIONS: These results suggest that both ETI and LMA may be acceptable choices for management of the airway in the helmeted athlete. Time to airway was significantly decreased with the use of LMA, regardless of the experience level of the intubator. Lighting conditions had no effect on success. PMID- 21617527 TI - Serum levels of brain-derived neurotrophic factor and cortisol to sulfate of dehydroepiandrosterone molar ratio associated with clinical response to L theanine as augmentation of antipsychotic therapy in schizophrenia and schizoaffective disorder patients. AB - OBJECTIVES: L-Theanine (gamma-glutamylethylamide) augmentation to antipsychotic therapy ameliorates positive, activation, and anxiety symptoms in schizophrenia and schizoaffective disorder patients. This study examines the association between circulating levels of neurochemical indicators and the beneficial clinical effects of L-theanine augmentation. METHODS: Serum levels of neurochemical indicators such as brain-derived neurotrophic factor (BDNF), dehydroepiandrosterone (DHEA), its sulfate (DHEAS), cortisol, cholesterol, and insulin were monitored in 40 schizophrenia and schizoaffective disorder patients during an 8-week, double-blind, randomized, placebo-controlled trial with L theanine (400 mg/d). Multiple regression analysis was applied for searching association between improvement in symptom scores and changes in circulating levels of neurochemical indicators for an 8-week trial. RESULTS: Regression models among L-theanine-treated patients indicate that circulating levels of BDNF and cortisol-to-DHEAS*100 molar ratio were significantly associated with the beneficial clinical effects of L-theanine augmentation. Variability of serum BDNF levels accounted for 26.2% of the total variance in reduction of dysphoric mood and 38.2% in anxiety scores. In addition, the changes in cortisol-to-DHEAS*100 molar ratio accounted for 30% to 34% of the variance in activation factor and dysphoric mood scores and for 15.9% in anxiety scores. Regression models among placebo-treated patients did not reach significant level. CONCLUSIONS: These preliminary results indicate that circulating BDNF and cortisol-to-DHEAS*100 molar ratio may be involved in the beneficial clinical effects of L-theanine as augmentation of antipsychotic therapy in schizophrenia and schizoaffective disorder patients. PMID- 21617530 TI - Genotype of lattice corneal dystrophy (R124C mutation in TGFBI) in a patient presenting with features of Avellino corneal dystrophy. PMID- 21617528 TI - Music recognition in frontotemporal lobar degeneration and Alzheimer disease. AB - OBJECTIVE: To compare music recognition in patients with frontotemporal dementia, semantic dementia, Alzheimer disease, and controls and to evaluate the relationship between music recognition and brain volume. BACKGROUND: Recognition of familiar music depends on several levels of processing. There are few studies about how patients with dementia recognize familiar music. METHODS: Subjects were administered tasks that assess pitch and melody discrimination, detection of pitch errors in familiar melodies, and naming of familiar melodies. RESULTS: There were no group differences on pitch and melody discrimination tasks. However, patients with semantic dementia had considerable difficulty naming familiar melodies and also scored the lowest when asked to identify pitch errors in the same melodies. Naming familiar melodies, but not other music tasks, was strongly related to measures of semantic memory. Voxel-based morphometry analysis of brain magnetic resonance imaging showed that difficulty in naming songs was associated with the bilateral temporal lobes and inferior frontal gyrus, whereas difficulty in identifying pitch errors in familiar melodies correlated with primarily the right temporal lobe. CONCLUSIONS: The results support a view that the anterior temporal lobes play a role in familiar melody recognition, and that musical functions are affected differentially across forms of dementia. PMID- 21617531 TI - Maturation of the occlusion effect: a bone conduction auditory steady state response study in infants and adults with normal hearing. AB - OBJECTIVE: The aim of this study was to investigate the maturational time course of the occlusion effect in infants with normal hearing. The objectives were (i) to investigate the occlusion effect in a larger group of young infants, (ii) to determine whether the occlusion effect is seen in bone conduction auditory steady state responses (ASSRs) for older infants, and (iii) to investigate the mechanisms that underlie bone conduction hearing in unoccluded and occluded ears in infants by measuring sound pressure in the ear canal. DESIGN: Experiments 1A and 1B: The SPL in the ear canal to 500, 1000, and 2000 Hz bone-conducted pure tones were compared in 22 young infants (0-7 mo), 10 older infants (10-22 mo), and 34 adults, all with normal hearing, for unoccluded and occluded ears. Experiment 2: Bone conduction behavioral thresholds in 17 adults were compared for unoccluded and occluded ears at 500, 1000, 2000, and 4000 Hz. Experiment 3: Bone conduction ASSR thresholds and amplitudes were compared in 22 young infants, 10 older infants, and 20 adults for an unoccluded and occluded test ear. Stimuli were bone-conducted amplitude/frequency-modulated tones presented simultaneously at 500, 1000, 2000, and 4000 Hz. RESULTS: There were significant increases in sound pressure in the ear canal for stimuli presented at 40 dB HL when ears were occluded at 500 and 1000 Hz for all age groups. Infants showed the largest increases in SPL at 500 and 1000 Hz (5-8 dB > adults). Young infants showed no significant decreases in ASSR thresholds (2-6 dB) and amplitudes (0-10 nV) across frequency with occlusion; however, a significant number of infants had an occlusion effect at 500 Hz. Older infants showed a nonsignificant decrease in ASSR thresholds with occlusion (8 dB), a significant increase in ASSR amplitudes at 1000 Hz (6-21 nV), and a significant number of infants with an occlusion effect at 1000 Hz. Adult behavioral thresholds decreased significantly when ears were occluded at 500 and 1000 Hz; for ASSRs, thresholds also decreased (6-7 dB) and amplitudes increased (3-11 nV) at both 500 and 1000 Hz, but the mean trends and statistical findings were not in agreement in all cases. A significant number of adult subjects had an occlusion effect at 500 and 1000 Hz for both behavioral and ASSR thresholds. CONCLUSIONS: Our findings suggest that the occlusion effect for ASSR thresholds in young infants is small but emerging at 500 Hz but negligible at 1000 Hz and that the occlusion effect in older infants is emerging at both 500 and 1000 Hz. The clinical implications of these findings are that it is appropriate to conduct bone conduction testing on young infants without compensating for an occlusion effect; however, for older infants, it is prudent to remove insert earphones during bone conduction testing. For both young and older infants, occluding the ear canal increases the sound pressure near the tympanic membrane; however, this pathway appears to contribute less to bone conduction hearing when ears are occluded compared with adults as measured by ASSRs. PMID- 21617532 TI - Performance of enhanced liver fibrosis plasma markers in asymptomatic individuals with ZZ alpha1-antitrypsin deficiency. AB - OBJECTIVES: Alpha1-antitrypsin deficiency (AATD) is a common genetic cause of chronic liver disease. According to retrospective studies, up to 25% of those with homozygous ZZ (Glu 342 to Lys) AATD suffer from liver cirrhosis and/or liver cancer in late adulthood. We hypothesized that the plasma markers for liver fibrosis, necrosis, and apoptosis may identify AATD individuals at higher risk for liver diseases. METHODS: The study cohort included 52 clinically healthy ZZ AATD individuals of 34 years of age, identified in the Swedish neonatal screening of 1972-1974, and 81 age-matched controls with normal MM AAT variant. We analyzed plasma levels of the enhanced liver fibrosis (ELF) panel, including plasma tissue inhibitor of metalloprotease-1, amino-terminal propeptide of type III collagen and hyaluronic acid (HA), and the M30 and M65 antigens, markers for apoptosis/necrosis. RESULTS: Higher levels of tissue inhibitor of metalloprotease 1 (52%, P<0.001), amino-terminal propeptide of type III collagen (12%, P<0.05), HA (17% not significant), and M65 (13.4%, P=0.043) were found in ZZ than in MM patients. In the ZZ group, plasma levels of AAT correlated with M65 (P<0.01) and with HA (P<0.05). On the basis of the ELF panel, M30 and M65, a logistic regression model enabled us to correctly classify 81.2% of the originally grouped ZZ and MM cases with a sensitivity of 73.1% and a specificity of 86.4%. CONCLUSION: The ELF markers are associated with ZZ AATD at early adulthood, and can be considered as a useful tool to identify ZZ cases at an increased risk of developing liver diseases later in life. PMID- 21617533 TI - Prevalence of hepatitis B in the southeast of China: a population-based study with a large sample size. AB - OBJECTIVE: Hepatitis B is a major public health problem in China. However, in the past 10 years, estimates of the prevalence of hepatitis B virus (HBV) have been sparse, with results that do not always agree. The purpose of this study was to investigate the epidemiology of HBV in adults and to provide the most recent baseline data for planning and monitoring of health. METHODS: The prevalence of HBV was determined in a community-based, cross-sectional, age-stratified sample of adults (aged 20 years or older). The study population was selected using random multistage cluster sampling. Demographic information and serological samples were obtained from 19 933 participants. RESULTS: The prevalence of anti HBV core antibody, HBV surface antigen (HBsAg), and anti-HBV surface antibody was 38.6, 7.9, and 44.9%, respectively. We found statistically significant differences in hepatitis B infection between men and women (P<0.001). The prevalence of HBV infection increased with age. The rates of HBsAg positivity and HBV infection in individuals living on an island were higher than those of individuals living in plains. The alanine aminotransferase level was elevated (>=38 IU/l) in 15.7 and 7.2% of HBsAg-positive and HBsAg-negative patients; the proportion of elevated alanine aminotransferase was markedly higher in men (12.7%) than in women (4.7%) (P<0.001). The alpha-fetoprotein-positive rate was 2% in HBsAg-positive individuals who were aged 30 years or older. CONCLUSION: Borderline intermediate/high endemicity was found among adults in the northern region of Zhejiang Province. The vaccination program has contributed to a reduction in infection over the last 20 years, although further efforts are required to turn the region into a 'low-endemicity' area. PMID- 21617534 TI - Ultraviolet radiation as a risk factor for cataract and macular degeneration. AB - The human eye is constantly exposed to sunlight and artificial lighting. Light transmission through the eye is fundamental to its unique biological functions of directing vision and circadian rhythm, and therefore, light absorbed by the eye must be benign. However, exposure to the intense ambient radiation can pose a hazard particularly if the recipient is over 40 years of age. This radiation exposure can lead to impaired vision and transient or permanent blindness.Both ultraviolet-A (UV-A) and UV-B induce cataract formation and are not necessary for sight. Ultraviolet radiation is also a risk factor for damage to the retinas of children. The removal of these wavelengths from ocular exposure will greatly reduce the risk of early cataract and retinal damage. One way this may be easily done is by wearing sunglasses that block wavelengths below 400 nm (marked 400 on the glasses). However, because of the geometry of the eye, these glasses must be wraparound sunglasses to prevent reflective UV radiation from reaching the eye. Additional protection may be offered by contact lenses that absorb significant amounts of UV radiation.In addition to UV radiation, short blue visible light (400-440 nm) is a risk factor for the adult human retina. This wavelength of light is not essential for sight and not necessary for a circadian rhythm response. For those over 50 years old, it would be of value to remove these wavelengths of light with specially designed sunglasses or contact lenses to reduce the risk of age-related macular degeneration. PMID- 21617535 TI - Brevundimonas diminuta keratitis. AB - PURPOSE: To report the first documented case of keratitis secondary to the gram negative bacillus Brevundimonas diminuta. METHODS: A 66-year-old woman with an ocular history of bilateral soft contact lens wear presented with a large, 6*9 mm, culture-positive Pseudomonas aeruginosa-induced corneal ulcer. After weeks of therapy, she developed secondary keratitis. Repeat cultures grew a rare, gram negative bacillus, B. diminuta. RESULTS: Targeted therapy followed by adjustments based on repeat bacterial cultures with sensitivities resulted in slow resolution of the ulcer. After months of therapy, the cornea recovered leaving only a minimal corneal scar. Visual acuity improved from a low of Hand Motions to a final 20/40. CONCLUSIONS: This is the first reported case of ocular infection with B. diminuta. This case demonstrates the need to be aware of potential rare secondary infections in corneas compromised by an initial infection and to adjust pharmacotherapy based on sensitivities and clinical signs. PMID- 21617536 TI - Meibomian gland therapeutic expression: quantifying the applied pressure and the limitation of resulting pain. AB - OBJECTIVES: The purposes of this study were to determine (1) the pressure required to express the first nonliquid material from nonfunctional lower lid meibomian glands, (2) the pressure required to evacuate all of the expressible material from the glands (simulating the authors' methodology for therapeutic meibomian gland expression), and (3) the level of pain associated with these procedures. METHODS: All patients (n=28) were recruited from those presenting for ocular examinations at a single practice. Custom instrumentation exerting pressures from 1.0 to 150.0 psi was developed to quantify the pressure applied during expression. The instrument was applied to the inner surface of the lower lid. The lid was then compressed between the thumb and the contact surface of the instrument. The applied pressure was displayed on a digital meter. The first procedure evaluated the pressure required to obtain the first nonliquid material from nonfunctional glands. The second evaluated the pressure required for evacuating all expressible gland contents. The pain response was monitored throughout the procedure. RESULTS: The pressure to obtain the first nonliquid material ranged from 5 to 40 psi (mean=16.1+/-8.2 psi) and for the evacuation of expressible contents, from 10 to 40 psi (mean=25.6+/-11.4 psi). Only 7% of the patients could tolerate the pressure necessary to administer complete therapeutic expression along the entire lower eyelid. CONCLUSIONS: Forces of significant magnitude are required for therapeutic expression. Pain is the limiting factor for the conduct of this treatment. PMID- 21617537 TI - Effects of peroxide-based contact lens-disinfecting systems on human corneal epithelial cells in vitro. AB - OBJECTIVE: To evaluate the effects of residual hydrogen peroxide (H2O2) in neutralized H2O2-based contact lens-disinfecting solutions on morphology, viability, and barrier function of monolayer and stratified human corneal-limbal epithelial (HCLE) cells. METHODS: Cells were exposed to contact lens formulations containing 0.01% H2O2 for 10, 20, or 60 minutes. The morphology of monolayer or stratified cells was observed by microscopy. Monolayer or stratified cell viability was determined using a live/dead assay, and monolayer cell viability was quantified using flow cytometry. Effects of formulations on barrier function of stratified HCLE cells were evaluated by measuring fluorescein permeability and transepithelial resistance of cultures grown on membrane inserts. To determine the sensitivity of the tests to peroxide damage, stratified cells were also exposed to 0.01% to 0.3% H2O2 in culture medium. RESULTS: All formulations caused swelling of monolayer cells. Formulations with or without H2O2 at pH 7.9 caused mild decreases in monolayer cell viability but did not affect viability or barrier function of stratified HCLE cells. H2O2 (>=0.1%) in culture medium caused damage without recovery to stratified HCLE cells. CONCLUSIONS: Although tests on stratified cells are capable of detecting damage caused by H2O2 in culture medium, residual H2O2 in neutralized ophthalmic formulations had no effect on stratified cells in vitro. These data suggest that H2O2, used appropriately, is a safe disinfectant. Data comparing monolayer and stratified cultures suggest that monolayers are more sensitive to peroxide damage and that the effects of neutralized formulations on stratified cells may better predict the intact corneal epithelial response. PMID- 21617538 TI - Case report: spontaneous Stenotrophomonas maltophilia keratitis in a diabetic patient. AB - OBJECTIVES: To report a rare case of spontaneous Stenotrophomonas maltophilia keratitis in a diabetic patient. METHODS: A case report of this rare ulcerative keratitis case with an extensive review of the literature. RESULTS: The patient's corneal infiltrate was successfully cultured and a rare ocular organism identified. The patient's presentation and course and the response to empiric treatment are discussed. CONCLUSIONS: Consideration of the antibiotic combination chosen here can be considered in the treatment of S. maltophilia keratitis after appropriate cultures are taken and the organism isolated. PMID- 21617539 TI - Early opacification and subsequent in vivo clearing of a silicone intraocular lens. AB - OBJECTIVE: To report a case of early opacification and spontaneous in vivo clearing of a silicone intraocular lens (IOL). METHODS: A 79-year-old women underwent uncomplicated cataract extraction and subsequent implantation of a +21.5 D AMO Z9002 (Abbott Medical Optics, Santa Ana, CA) silicone lens. RESULTS: On postoperative day 1, the patient presented with a translucent, milky white appearing, diffuse IOL opacification. On postoperative day 8, the IOL periphery had cleared, but a full thickness haze persisted in the central portion of the optic. By postoperative day 14, the IOL had cleared. To date, the lens has remained clear. CONCLUSION: One possible mechanism might be exposure to industrial chemicals; however, the manufacturer reports no other similar lens opacities from this manufacturing batch of IOLs. We recommend that surgeons who implant or have implanted an AMO Z9002 silicone IOL and observe similar findings follow the patient closely for resolution of the haze. PMID- 21617540 TI - Technetium-99m-labeled RBC scintigraphy: less relative utility now than ever? PMID- 21617541 TI - Predictors of recurrence after resection of small gastric gastrointestinal stromal tumors of 5 cm or less. AB - GOALS: To evaluate the recurrence predicting factors of small gastric gastrointestinal stromal tumors (GISTs) through the long-term follow-up after surgical/endoscopic resection. BACKGROUND: Although small gastric GISTs are known to have a low risk of recurrence after complete resection, the prognostic factors are not well known. STUDY: The study retrospectively analyzed the records of 136 patients with primary gastric GISTs of 5 cm or less without metastasis who underwent surgical/endoscopic resection between March 1997 and December 2008 at the Asan Medical Center, and who were followed-up for at least 3 months after resection. Specimens were assessed for tumor size, mitotic index, and microscopic resection margin. Specimen sections were immunohistochemically stained to determine the levels of expression of the cell cycle proteins p53, p16(INK4), pRb, cyclin D1, and Ki-67. DNA was extracted from high-risk tumors to analyze for KIT mutations. RESULTS: Among 136 patients, 5 (3.7%) patients with tumors with a high mitotic index showed recurrence at a median 23 months post resection. None of 14 patients with microscopic positive resection margins showed recurrence during a median follow-up time of 32 months. A high mitotic index was a predictor of recurrence (P<0.001), but that tumor size, method of resection, or margin status were not. In addition, abnormal p53 expression was found to be associated with recurrence (P=0.004). All assessable high-risk tumors had a KIT exon 11 mutation. CONCLUSIONS: Predictors of recurrence of gastric GISTs of 5 cm or less were a high mitotic index and abnormal p53 expression. A positive microscopic resection margin was not associated with recurrence. PMID- 21617542 TI - Comparison of clevudine and entecavir for treatment-naive patients with chronic hepatitis B virus infection: two-year follow-up data. AB - BACKGROUND/AIM: Clevudine and entecavir are highly potent antiviral agents being used in treatment of chronic hepatitis B. However, no data comparing clinical efficacy and safety of these 2 drugs over a long-term period is available. The aims of this study are to compare virologic, biochemical, and serologic response rates of clevudine and entecavir, as well as treatment failure rates up to 2 years. METHODS: Data of patients who started clevudine (n = 86) or entecavir (n = 159) as a primary treatment for chronic hepatitis B at Korea University Ansan or Guro Hospital between January 2007 and June 2008 were analyzed. RESULTS: Treatment responses were compared at 3-month intervals up to 24 months. Per protocol analysis showed no difference in virologic responses between the 2 groups at all time points, except at 18 months. When analyzed on intention-to treat basis for virologic response at 24 months, the response rates were 45.3% in the clevudine group and 72.3% in the entecavir group, which are significantly different (P < 0.001). Rates of biochemical response and HBeAg seroconversion were not significantly different between the groups at all time points. Up to 24 months, antiviral resistance developed in 18 patients (24.4%) in the clevudine group. Clevudine was discontinued owing to muscle-related problems in 10 patients (11.6%). CONCLUSIONS: Although both drugs showed potent antiviral activity, entecavir showed better virologic response at 24 months, primarily owing to treatment failures in the clevudine group that were associated with development of drug resistance and muscle-related problems. PMID- 21617544 TI - Probiotics in functional bowel disorders getting it right. PMID- 21617543 TI - Prevalence of Barrett esophagus in first-degree relatives of patients with esophageal adenocarcinoma. AB - AIM: The aim of this study is to determine the prevalence of Barrett esophagus (BE) in first-degree relatives of patients with esophageal adenocarcinoma (EAC) and Barrett high-grade dysplasia (HGD). METHODS: After Institutional Review Board approval, first-degree relatives of patients with EAC/HGD underwent unsedated ultrathin transnasal endoscopy (UUTNE) with biopsy. BE was suspected if any salmon-colored epithelial tongues were seen above the gastroesophageal junction. A diagnosis of BE was made only if biopsy from these areas confirmed columnar lined epithelium with intestinal metaplasia. RESULTS: From 23 families, 47 first degree relatives underwent ultrathin transnasal endoscopy and 1 patient underwent routine upper endoscopy with sedation as part of this study. The mean age of cases was 44.4 years. All patients tolerated the procedure well and there were no procedure-related complications. BE was suspected in 16 (34%) patients and confirmed in 13 of 16 (27.7%) patients. There were 4 long segments (>3 cm) and 9 short segments (<3 cm) of BE. CONCLUSION: There is a significantly higher than expected prevalence of BE in first-degree relatives of patients with EAC/HGD. This should be taken in to consideration to develop further screening guidelines. Further work is needed to confirm these findings. Unsedated transnasal endoscopy is a safe and well-tolerated method for BE screening. PMID- 21617545 TI - A case of levamisole-induced systemic vasculitis and cocaine-induced midline destructive lesion: a case report. AB - We describe a case of antineutrophil cytoplasmic antibody-positive vasculitis from levamisole-tainted cocaine with concomitant cocaine-induced midline destructive lesions of the palate and nasal septum. The diagnosis was confirmed after extensive clinical, laboratory, pathologic, and radiographic testing. Timely recognition of this clinical entity is critical to avoid misdiagnosis and unnecessary treatment with potentially harmful cytotoxic agents. Given the high rate of levamisole contamination within the nation's cocaine supply, clinicians should be alerted to this emerging health threat. PMID- 21617546 TI - Listen to the axillary artery: diagnosis of occult giant cell arteritis. AB - Large-vessel involvement is increasingly recognized in giant cell arteritis (GCA) and frequently occurs with an unspecific systemic inflammatory syndrome or fever of unknown origin. We describe the case of a 68-year-old woman with a history of polymyalgia rheumatica who presented with progressive weight loss, unexplained anemia, and a marked humoral inflammatory response. Diagnosis of large-vessel GCA was facilitated by axillary artery auscultation, revealing a bilateral axillary artery bruit. The diagnosis was confirmed by color duplex sonography and temporal artery biopsy. In elderly patients with an unspecific systemic inflammatory condition, axillary artery auscultation is a simple but valuable tool for detection of underlying occult large-vessel GCA. PMID- 21617547 TI - Reversal of transfusion dependence by tumor necrosis factor inhibitor treatment in a patient with concurrent rheumatoid arthritis and primary myelofibrosis. AB - Tumor necrosis factor alpha (TNF-alpha) is a proinflammatory cytokine with pleiotropic effects. Currently, TNF-alpha inhibitors are approved by the Food and Drug Administration for a number of diseases including rheumatoid arthritis, juvenile idiopathic arthritis, psoriatic arthritis, ankylosing spondylitis, Crohn disease, and ulcerative colitis. We report a patient with seropositive rheumatoid arthritis with concurrent primary myelofibrosis, who had transfusion-dependent anemia and moderate thrombocytopenia that reversed during treatment with adalimumab. Rheumatoid arthritis and myeloproliferative disorder or myelodysplastic disorder often coexist, and treatment with standard immunosuppressants becomes complex. This report adds to the accumulating evidence of the safety of TNF-alpha inhibitors when primary myelofibrosis is present and generates discussion for further exploration of the potential therapeutic benefit of TNF-alpha inhibitors in cytopenias associated with primary myelofibrosis. PMID- 21617548 TI - Prevalence and progress of joint symptoms in hereditary hemochromatosis and symptomatic response to venesection. PMID- 21617549 TI - Risk factors associated with depressive symptoms in Venezuelan patients with rheumatoid arthritis. PMID- 21617550 TI - A case of immunoglobulin 4-related disease with bilateral mass-forming lesions in the nasolacrimal ducts. AB - This case study describes a 58-year-old man with watery eyes, bilateral swelling of the ala nasi and submandibular glands, and a swollen right parotid gland. Computed tomography revealed mass-forming lesions in both nasolacrimal ducts, extending bilaterally to the lacrimal sac and inferior meatus in the nose. Pathologic investigation showed marked infiltrates of immunoglobulin 4 (IgG4) positive plasma cells in the nasolacrimal duct lesion, which led us to diagnose IgG4-related disease. Oral prednisolone improved the symptoms of watery eyes and the bilateral swelling of the nasolacrimal duct and salivary gland enlargement. To the best of our knowledge, this is the first report of IgG4-related disease involving mass-forming lesions in the nasolacrimal ducts. PMID- 21617551 TI - Oligosymptomatic herpetic hepatitis in a patient with rheumatoid arthritis using corticosteroid and methotrexate. PMID- 21617552 TI - Febuxostat efficacy in allopurinol-resistant tophaceous gout. AB - Allopurinol has been a standard hypouricemic agent for more than 40 years, but febuxostat, which is also a xanthine oxidase inhibitor, is now available. We present a case of tophaceous gout with uric acid levels that did not respond to prescription of very high doses of allopurinol or uricosuric agents. With febuxostat treatment, the patient had rapid and sustained reduction in her uric acid levels to target range. This case raises the possibility of currently unrecognized pharmacologic differences between allopurinol and febuxostat. PMID- 21617553 TI - Rapid development of sarcoid tenosynovitis. AB - We report a case of acute tenosynovitis from sarcoidosis and review previously reported cases of this entity. A woman with known pulmonary sarcoidosis rapidly developed painless nodules in the tendon sheaths of the dorsum of both hands and wrists. Sarcoid tenosynovitis is almost exclusively found in the upper extremity. The flexor and extensor tendons are equally affected. The condition may respond to medical therapy including corticosteroids and other immunomodulating medications. Surgical debulking and tendon sheathectomy have also been curative. The disease has also been reported to spontaneously resolve. Our patient dramatically improved while on methotrexate. PMID- 21617554 TI - Double-blind, randomized, controlled, pilot study comparing classic ayurvedic medicine, methotrexate, and their combination in rheumatoid arthritis. AB - OBJECTIVE: To compare classic Ayurveda, methotrexate (MTX), and their combination in a double-blind, randomized, double-dummy, pilot trial in rheumatoid arthritis (RA) for 36 weeks. METHODS: Forty-three seropositive RA patients by American College of Rheumatology (ACR) criteria with disease duration of less than 7 years were assigned to the following treatment groups: MTX plus Ayurvedic placebo (n = 14), Ayurveda plus MTX placebo (n = 12), or Ayurveda plus MTX (n = 17). Outcomes included the Disease Activity Score (DAS28-CRP), ACR20/50/70, and Health Assessment Questionnaire--Disability Index. All measures were obtained every 12 weeks for 36 weeks. Analyses included descriptive statistics, analysis of variance, chi2, or Student t test. The unique features of this study included the development of placebos for each Ayurvedic pharmacological dosage form and individualization of Ayurvedic therapy. RESULTS: All groups were comparable at baseline in demographics and disease characteristics. There were no statistically significant differences among the 3 groups on the efficacy measures. ACR20 results were MTX 86%, Ayurveda 100%, and combination 82%, and DAS28-CRP response were MTX -2.4, Ayurveda -1.7, and combination -2.4. Differences in adverse events among groups were also not statistically significant, although the MTX groups experienced more adverse event (MTX 174, Ayurveda 112, combination 176). No deaths occurred. CONCLUSIONS: In this first-ever, double-blind, randomized, placebo-controlled pilot study comparing Ayurveda, MTX, and their combination, all 3 treatments were approximately equivalent in efficacy, within the limits of a pilot study. Adverse events were numerically fewer in the Ayurveda-only group. This study demonstrates that double-blind, placebo-controlled, randomized studies are possible when testing individualized classic Ayurvedic versus allopathic treatment in ways acceptable to western standards and to Ayurvedic physicians. It also justifies the need for larger studies. PMID- 21617555 TI - Outcome and predictors of kidney disease progression in Puerto Ricans with systemic lupus erythematosus initially presenting with mild renal involvement. AB - OBJECTIVES: The aims of this study were to determine the outcomes and predictors of renal disease progression in Puerto Ricans with systemic lupus erythematosus (SLE) initially presenting mild renal involvement. METHODS: A retrospective cohort of 61 patients with SLE (per American College of Rheumatology classification) with mild renal involvement was studied. Mild renal disease was defined as glomerular filtration rate (GFR) of 90 mL/min or higher in the presence of proteinuria (>0.25 g/d, but <3.5 g/d), hematuria, and/or urinary cellular casts. Demographic parameters, clinical manifestations, serologic markers, comorbidities, pharmacologic treatments, disease activity, and damage accrual were determined at onset of renal disease. Factors associated with renal disease progression were evaluated using recurrent event survival analysis. RESULTS: Of 61 patients, 55 (90.2%) were women. The mean (SD) age at renal onset was 29 (11.2) years, and the mean (SD) follow-up period was 5.1 (3.4) years. Thirty-eight patients had a decline in GFR. Thirty-two had a mild decline (GFR = 60-89 mL/min), 5 developed moderate to severe renal insufficiency (GFR = 15-59 mL/min), and 1 evolved to end-stage renal disease (GFR < 15 mL/min). In the Cox model, low C4 levels and proteinuria greater than 0.5 g/d were associated with an earlier decline in GFR. CONCLUSIONS: Most Puerto Rican patients with SLE initially presenting with mild renal involvement had a decrease in GFR after an average of 5 years of kidney disease, although most had a mild dysfunction. Low C4 levels and proteinuria were predictors of an earlier decline in GFR. We emphasize that awareness of these factors may contribute to early identification of individuals at risk for renal deterioration. PMID- 21617556 TI - Nerve blocks at the wrist for painful injections of the palm. AB - PURPOSE: Injections into the palmar hand for trigger finger, palmar flexor tenosynovitis, and Dupuytren contracture can be very painful. This randomized, controlled study evaluated nerve block anesthesia at the wrist for prevention of procedural pain associated with painful injection of the palmar hand. METHODS: Forty-seven corticosteroid injections for trigger fingers in 19 individuals were randomized to (1) anesthesia consisting of median and ulnar nerve block with 1% lidocaine anesthesia followed by standard injection or (2) standard injection alone using the 1-needle 2-syringe technique consisting of transthecal dilation of the synovial sheath with 0.5 mL 1% lidocaine with a mechanical syringe, the reciprocating procedure device, followed by injection with 20 mg triamcinolone acetonide. Baseline pain, needle insertion/sheath dilation pain, corticosteroid injection pain, resolution of trigger finger, and pain at outcome (2 weeks) were determined. RESULTS: Standard injection for trigger finger was associated with significant pain in 100% of subjects. Nerve blocks at the wrist provided effective anesthesia, resulting in a 56% reduction in injection pain compared with direct injection (P < 0.01). There was 100% resolution of trigger finger in both treatment groups. Pain at the 2-week outcome, reduction in pain from baseline, responders, and nonresponders were not statistically different (P > 0.3 for all). Eighty-eight percent of subjects preferred nerve block anesthesia to direct injection (P < 0.0001). CONCLUSIONS: Nerve block anesthesia at the wrist before palmar injection is preferred by patients and is highly effective in preventing pain associated with injection of the palmar hand for trigger finger and other painful hand procedures. PMID- 21617557 TI - Rheumatic complaints in women taking aromatase inhibitors for treatment of hormone-dependent breast cancer. AB - BACKGROUND: The rheumatic adverse effects accompanying treatment with aromatase inhibitors (AIs) in hormone-dependent breast cancer represent an area of clinical relevance and emerging concern. This report describes these rheumatic complaints detailing their clinical pattern. METHODS: During 1-year period, 18 consecutive postmenopausal women (mean age, 58.33 years; range, 52-66 years) in treatment with AIs for hormone-dependent breast cancer (mean duration of therapy, 12.0 months; range, 9.1-17.7 months) were referred for evaluation in the outpatient clinic of the rheumatology unit in relation to rheumatic complaints. According to a routine protocol planned with oncologists, patient evaluations consisted of a complete clinical examination with careful assessment of rheumatic complaints and related physical symptoms, followed by laboratory testing and a bone scintiscan. In no cases were rheumatic complaints present before AI therapy. RESULTS: On the basis of clinical data and investigations and by applying accepted diagnostic criteria, a diagnosis of an undifferentiated spondyloarthropathy was reached in 10 (55.5%) of the 18 patients studied, and an oligoarthritis was shown in 2 more patients (11.1%), whereas a simple arthralgia was found in the remaining 6 patients (33.3%). In the patients meeting criteria as belonging to a spondyloarthritic subset, a family history positive for psoriasis and celiac disease was shown in 2 and 1 instance, respectively, whereas HLA-CW6 and HLA-B27 were detected in 3 and 1 case. A high serum level of anti-cyclic citrullinated peptide antibodies was shown in 1 patient with oligoarthritis. Most of the patients (16/18) were treated with nonsteroidal anti-inflammatory drugs or with corticosteroids. Methotrexate (10 mg weekly) was added in 3 of these patients, nonresponders. Aromatase inhibitor discontinuation was needed in the remaining 2 cases with spontaneous resolution of symptoms over time. CONCLUSIONS: Data from the present study emphasize a previously unsuspected high prevalence of defined arthritides underlying these rheumatic complaints. Therefore, investigative efforts should be addressed to better clarify the clinical and pathogenetic significance of these important consequences of AI therapy. An accurate monitoring of rheumatic complaints has to be suggested to patients taking AI therapy, with a rapid referral to a rheumatologist in the case of consistent suspicion of an inflammatory arthritis. PMID- 21617558 TI - Five consecutive cases of a cutaneous vasculopathy in users of levamisole adulterated cocaine. AB - Five patients with an antineutrophil cytoplasmic antibody (ANCA)-associated cutaneous vasculopathy secondary to levamisole-adulterated cocaine were prospectively followed up at a single hospital. All patients presented with retiform purpura, with ear involvement being the most characteristic finding. Cocaine metabolites were present on urine toxicology screening, with 2 of 4 of those tested also being positive for levamisole. High-titer polyspecific ANCA and positive antiphospholipid antibody tests were defining laboratory features. Thrombosis and/or leukocytoclastic vasculitis were seen on skin biopsy. Improvement of skin lesions and laboratory findings occurred with cessation of cocaine; however, arthralgias and other complications developed. Levamisole adulterated cocaine is a cause of a cutaneous vasculopathy associated with characteristic laboratory and clinical features that allow it to be distinguished from classic ANCA-associated small-vessel vasculitides. The chronic sequelae of this syndrome and the potential role for immunosuppression are yet to be completely defined. PMID- 21617559 TI - The clinical scene investigator academy: the power of staff nurses improving patient and organizational outcomes. AB - Nurses are in a key position to have an impact on the national quality agenda. However, many lack the quality and safety methods to lead this work. The Clinical Scene Investigator Academy provided 23 nurses from 7 regional hospitals with the knowledge and support necessary to implement nurse-led quality improvement projects. Not only did each Clinical Scene Investigator project improve patient and/or staff outcomes but 3 such projects also had a significant economic impact. PMID- 21617560 TI - Validation and clinical utility of a bowel habit questionnaire in school-age children. AB - OBJECTIVES: The aim of the study was to validate a brief Bowel Habit Questionnaire (BHQ) with prospectively obtained data from a 14-day diary and to determine whether the BHQ predicts the development of medically significant constipation (MSC) during the following year. MATERIALS AND METHODS: The BHQ was distributed to parents of children ages 5 to 8 years during health supervision visits. Both the BHQ and subsequent diary were scored to indicate constipation if at least 2 of the following were reported: infrequent bowel movements, stool accidents, straining, avoidance, discomfort with defecation, or passing large stools >25% of the time. One year later, the BHQ was repeated to assess for MSC, defined as medical encounters about constipation or use of enemas, suppositories, laxatives, or stool softeners. RESULTS: MSC was reported for 57 (13.7%) of 416 children on the first BHQ. Paired BHQ and diary data were obtained for 269 children; 54 (20.1%) had diary scores indicating constipation. BHQ had a sensitivity of 59.6% (95% confidence interval [CI] 46.7%-71.4%) and a specificity of 82.6% (95% CI 77.0%-87.1%). One year later, 11 children (5.2%) had developed new-onset MSC; 7 (63.6%) of these children had initial BHQ scores of at least 2. Positive and negative predictive values for MSC were 19.4% (95% CI 9.8%-35.0%) and 97.7% (94.2%-99.1%), respectively. CONCLUSIONS: Parents often do not recognize constipation in young school-age children and most constipated children remain untreated. A brief screening questionnaire in this population proved to be valid but only moderately sensitive; efforts to improve sensitivity are needed before recommending it for routine use. PMID- 21617561 TI - Etiology of septicemia in children with acute leukemia: 9-year experience from a children's hospital in China. AB - Bloodstream infection is an important cause of death among leukemia patients, and the etiologic agent surveillance is important for the prophylaxis and treatment. This study aims to identify the common bloodstream isolates in hospitalized leukemia patients with septicemia in our hospital, to choose the ideal combination of antimicrobial agents for infection prophylaxis and to clarify the appropriate time for antibiotic prophylaxis. To know this information, a retrospective analysis was conducted over a 9-year period from July 2001 to July 2010 by reviewing medical records of leukemia children admitted to our hospital. The overall frequencies of isolation were 45% in Gram-positive bacteria, 53.8% in Gram-negative bacteria, and 1.2% in fungi, respectively. Coagulase-negative Staphylococci were the most common organisms isolated, accounting for 32.7% of the total blood culture isolates, followed by Escherichia coli (15.7%) and Klebsiella pneumoniae (7.1%). The incidence of septicemia caused by extended spectrum beta-lactamase-producing E. coli and K. pneumoniae was high (69.2% and 58.8% of total isolates, respectively). The coverage rate of antimicrobial combinations of "vancomycin+cefoperazone-sulbactam" and "vancomycin+piperacillin tazobactam" to blood culture isolates of leukemia patients in our hospital were 91.88% and 90.27%, respectively. More than 90% of septicemia occurred when the absolute neutrophil count was lower than 1.6*109/L and 83.05% when absolute neutrophil count was lower than 1.0*109/L. These results suggest that ongoing surveillance for antimicrobial susceptibility in leukemia children remains essential. Vancomycin+cefoperazone-sulbactam and vancomycin+piperacillin tazobactam are the good choice for leukemia children to prevent bacterial infections in our hospital. In an effort to reduce total consumption of antibiotics and to elevate the therapeutic efficacy, antibiotics prophylaxis should be started with the appearance of neutropenia in leukemia children. PMID- 21617562 TI - Parvovirus B19-associated hemophagocytic lymphohistiocytosis in a child with precursor B-cell acute lymphoblastic leukemia under maintenance chemotherapy. AB - Development of hemophagocytic lymphohistiocytosis (HLH) is quite rare among acute lymphoblastic leukemia (ALL) patients. We present a 3-year-old boy with precursor B-cell ALL, who was complicated by HLH because of parvovirus B19 infection during maintenance chemotherapy. Remarkable erythroid hypoplasia, giant normoblasts, and hemophagocytosed macrophages in bone marrow were important clues for the diagnosis. The patient was successfully treated with high-dose steroids and intravenous immunoglobulins. To our knowledge, this is the first report describing parvovirus B19-associated HLH in ALL. Our case highlights that parvovirus B19 can cause HLH, a potentially fatal disorder, and prolonged unexpected cytopenia in childhood ALL. PMID- 21617563 TI - Acute promyelocytic leukemia with complex translocation t(5;17;15)(q35;q21;q22): case report and review of the literature. AB - The t(15;17)(q22;q21), resulting in PML-RARA fusion gene, is a characteristic chromosomal translocation in acute promyelocytic leukemia (APL). We report a pediatric APL case with a 3-way translocation: t(5;17;15)(q35;q21;q22). Complete blood cell counts of a 12-year-old girl, of pale appearance, showed pancytopenia with increased blasts. Morphology and immunophenotype of the leukemic cells were compatible with APL. Karyotype analysis showed t(5;17;15)(q35;q21;q22) and add(7)(q32). We detected the PML-RARA fusion gene by both reverse transcriptase polymerase chain reaction and fluorescent in situ hybridization analysis. The patient underwent successful treatment with cytarabine with all-trans retinoic acid and anthracycline-based therapy. PMID- 21617564 TI - Congenital hemophagocytic lymphohistiocytosis in a preterm infant: cytokine profile and a review of the disease. AB - A preterm infant with very low birth weight was born with fetal onset familial hemophagocytic lymphohistiocytosis. Known gene abnormalities responsible for the disease were not identified in the patient. The infant died at 13 months of age owing to complications from cord blood stem cell transplantation. We found selectively elevated expression of interleukin-6 and chemokines in the cord blood of the patient. We also reviewed 7 other preterm cases of congenital hemophagocytic lymphohistiocytosis to highlight the significance of this condition, as it can cause ascites and hepatosplenomegaly in utero and be mistaken for congenital infection in the fetus. PMID- 21617565 TI - Squamous cell carcinoma of the lung in an autistic child who has never smoked. AB - Primary bronchogenic carcinoma of the lung is extremely rare in childhood, particularly the squamous cell type. Only 13 cases have been reported in the literature. We report a case of squamous cell carcinoma in an autistic, 16-year old boy who presented with a productive cough. Interestingly, he was a never smoker, but had been exposed to environmental tobacco smoking by his father for 13 years. The diagnosis was delayed by approximately 1 month due to his young age. He was diagnosed with squamous cell carcinoma of the lung by video-assisted thoracoscopic surgery, and chemotherapy was arranged. Considering his age, autism, and good performance status, a combined chemotherapeutic regimen with gemcitabine plus carboplatin was planned. After the second cycle of chemotherapy, the cough resolved and a computed tomography scan showed a partial response of the central conglomerated mass with the absence of the malignant pleural effusion. PMID- 21617566 TI - Asthma is a distinct comorbid condition in children with sickle cell anemia with elevated total and allergen-specific IgE levels. AB - Asthma in children with sickle cell anemia (SCA) is associated with increased morbidity and mortality. However, the definition of asthma in SCA is based on a physician's impression. In a retrospective cohort of children with SCA, relationships between a physician diagnosis of asthma and total and allergen specific immunoglobulin E levels were evaluated. In children with SCA, elevated total and specific immunoglobulin E levels were significantly associated with a diagnosis of asthma (P<0.05), further supporting the concept that asthma is a separate comorbid condition of SCA. PMID- 21617567 TI - Preservation of spinal instrumentation after development of postoperative bacterial infections in patients undergoing spinal arthrodesis. AB - STUDY DESIGN: This is a retrospective review. OBJECTIVE: The purpose of this study is to evaluate the efficacy of medical and surgical management of postoperative infections after the placement of spinal instrumentation in the context of retaining the instrumentation. SUMMARY OF BACKGROUND DATA: The implementation of spinal instrumentation for the treatment of various spinal conditions is associated with a risk of developing a postoperative wound infection. An aggressive medical and surgical approach is necessary for eradication of subfascial infections and treatment is often performed with the guidance of infectious diseases consultants. These medical specialists frequently recommend removal of the implanted spinal instrumentation due to the concern for the potential of persistent microbial infection resulting from the formation of bacteria harboring biofilm on the indwelling spinal hardware. METHODS: Of 854 consecutive patients who had undergone spinal instrumentation by the senior author, 17 patients were identified who developed postoperative infections, 16 of which were treated by the senior author. RESULTS: All patients underwent surgical debridement and received antimicrobial therapy in consultation with the infectious diseases consultants. Spinal instrumentation was preserved in all cases. Two patients had clinical and radiologic evidence of nonunion and later underwent a staged instrumentation revision procedure. Eradication of the infection was successful in all patients. The mean follow-up period was 2.1 years (range, 12 mo to 4.5 y). CONCLUSIONS: A combination of aggressive surgical debridement with microbial-guided pharmacotherapy enabled infection cure in all patients with postoperative bacterial infections after spinal instrumentation. Adjunct measures including the use of wound vacuum devices and long-term suppressive antimicrobial therapy were also used in high-risk patients. These results show that postoperative bacterial infections in the setting of spinal instrumentation can be successfully treated without removing titanium alloy instrumentation. PMID- 21617568 TI - Comparison of disc and vertebral wedging between patients with adolescent idiopathic scoliosis and Chiari malformation-associated scoliosis. AB - STUDY DESIGN: A comparative study. OBJECTIVE: To investigate the difference of disc and vertebral wedging between patients with adolescent idiopathic scoliosis (AIS) and Chiari malformation-associated scoliosis (CMS). SUMMARY OF BACKGROUND DATA: The wedging of disc and vertebral wedging of AIS are well investigated. However, the comparison of that wedging between AIS and CMS is not found in the literature. METHOD: This study included 35 cases of AIS and 31 cases of CMS. The disc and vertebral wedging was measured by Cobb method. The percentage of single disc and vertebral wedging in the whole thoracic or lumbar curve was calculated. The difference between disc and vertebral wedging, the apical vertebra and average adjacent vertebrae wedging, the apical discs and adjacent discs wedging in the same curve location, same diagnosis and similar Cobb angle group was compared. The difference of disc or vertebral wedging between AIS and CMS, Cobb angle <60 degrees and >=60 degrees was also compared. The correlation between the apical vertebral wedging angle and Cobb angle was performed by Pearson correlation analysis. RESULTS: The difference between disc and vertebral wedging in the same curve location, same diagnosis, and similar Cobb angle group was statistically significant (P<0.05). No significant difference of the disc or vertebral wedging was found between AIS and CMS in the same curve location and the similar Cobb angle group (P>0.05). No significant difference of the disc or vertebral wedging was found between Cobb angle <60 degrees and >=60 degrees groups in the same curve location and the same diagnosis (P>0.05). There was a positive correlation between the apical vertebra wedging angle and Cobb angle in AIS patients and CMS patients. The wedging of apical vertebra and disc was more than that in the adjacent disc and vertebra in the same curve. CONCLUSIONS: The relative wedging of intervertebral disc and vertebrae with AIS patients is similar to that of scoliosis with known cause (CMS). The wedging of discs and vertebrae in AIS patients may be an adaptive change secondary to some extravertebral factors. PMID- 21617569 TI - Quality of data in perinatal population health databases: a systematic review. AB - BACKGROUND: Administrative or population health datasets (PHDS) are increasingly being used for research related to maternal and infant health. However, the accuracy and completeness of the information in the PHDS is important to ensure validity of the results of this research. OBJECTIVE: To compile and review studies that validate the reporting of conditions and procedures related to pregnancy, childbirth, and newborns and provide a tool of reference for researchers. METHODS: A systematic search was conducted of Medline and EMBASE databases to find studies that validated routinely collected datasets containing diagnoses and procedures related to pregnancy, childbirth, and newborns. To be included datasets had to be validated against a gold standard, such as review of medical records, maternal interview or survey, specialized register, or laboratory data. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and/or kappa statistic for each diagnosis or procedure code were calculated. RESULTS: Forty-three validation studies were included. Under-enumeration was common, with the level of ascertainment increasing as time from diagnosis/procedure to birth decreased. Most conditions and procedures had high specificities indicating few false positives, and procedures were more accurately reported than diagnoses. Hospital discharge data were generally more accurate than birth data, however identifying cases from more than 1 dataset further increased ascertainment. CONCLUSIONS: This comprehensive collection of validation studies summarizing the quality of perinatal population data will be an invaluable resource to all researchers working with PHDS. PMID- 21617570 TI - Effect of patient selection method on provider group performance estimates. AB - BACKGROUND: Performance measurement at the provider group level is increasingly advocated, but different methods for selecting patients when calculating provider group performance have received little evaluation. OBJECTIVE: We compared 2 currently used methods according to characteristics of the patients selected and impact on performance estimates. RESEARCH DESIGN, SUBJECTS, AND MEASURES: We analyzed Medicare claims data for fee-for-service beneficiaries with diabetes ever seen at an academic multispeciality physician group in 2003 to 2004. We examined sample size, sociodemographics, clinical characteristics, and receipt of recommended diabetes monitoring in 2004 for the groups of patients selected using 2 methods implemented in large-scale performance initiatives: the Plurality Provider Algorithm and the Diabetes Care Home method. We examined differences among discordantly assigned patients to determine evidence for differential selection regarding these measures. RESULTS: Fewer patients were selected under the Diabetes Care Home method (n=3558) than the Plurality Provider Algorithm (n=4859). Compared with the Plurality Provider Algorithm, the Diabetes Care Home method preferentially selected patients who were female, not entitled because of disability, older, more likely to have hypertension, and less likely to have kidney disease and peripheral vascular disease, and had lower levels of predicted utilization. Diabetes performance was higher under Diabetes Care Home method, with 67% versus 58% receiving >1 A1c tests, 70% versus 65% receiving >=1 low density lipoprotein (LDL) test, and 38% versus 37% receiving an eye examination. CONCLUSIONS: The method used to select patients when calculating provider group performance may affect patient case mix and estimated performance levels, and warrants careful consideration when comparing performance estimates. PMID- 21617572 TI - Risk factors for household transmission of community-associated methicillin resistant Staphylococcus aureus. AB - BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) has emerged as a community pathogen. Community-associated (CA) MRSA infections have occurred among multiple members of a household. We describe the incidence of and risk factors for MRSA colonization among household contacts of children with CA-MRSA infections. METHODS: MRSA-infected children <18 years of age who lacked established healthcare-associated MRSA risk factors were identified through surveillance at 12 Minnesota hospital laboratories. Nasal swab specimens and information on medical history and hygiene behaviors were collected from case patients and enrolled household contacts during home visits. S. aureus isolates obtained from nasal cultures were screened for oxacillin resistance. RESULTS: In all, 236 households consisting of 236 case-patients and 712 household contacts were enrolled. Home visits were conducted on an average of 69 days after the onset of symptom in case-patients (range: 16-178 days). Twenty-nine (13%) case patients and 82 (12%) household contacts had MRSA nasal colonization. Nasal MRSA colonization in >= 1 household contact occurred in 58 (25%) households. Household contacts who assisted the case-patient to bathe or who shared balms/ointments/lotion with the case-patient were more likely to be colonized (P < 0.01, P < 0.05), whereas those who reported using antibacterial versus nonantibacterial soap for hand washing were less likely to be colonized (P < 0.05) with MRSA clonally related to the case-patient infection isolate. CONCLUSIONS: Only 13% of case-patients had MRSA nasal colonization on an average of 69 days after their initial MRSA infection. CA-MRSA colonization may be short lived or may occur at non-nasal sites. One quarter of households had at least one household contact colonized with MRSA. Modifiable behaviors, such as sharing personal items, may contribute to transmission. PMID- 21617573 TI - Immunogenicity and safety of an inactivated hepatitis A vaccine when coadministered with measles-mumps-rubella and varicella vaccines in children less than 2 years of age. AB - BACKGROUND: Hepatitis A vaccination in early childhood has reduced hepatitis A transmission. Coadministration of hepatitis A vaccine with other childhood vaccines may assist completion of the age-appropriate immunization schedule. We assessed the immunogenicity and safety of an inactivated hepatitis A virus vaccine when coadministered with measles-mumps-rubella (MMR) and varicella vaccines in children less than 2 years of age. METHODS: In this open-label, randomized, multicenter study, 3 groups of healthy children 15 months of age received either 2 doses of hepatitis A vaccine 6 to 9 months apart (n = 324), hepatitis A vaccine coadministered with MMR and varicella vaccines and a second dose of hepatitis A vaccine 6 to 9 months later (n = 462), or MMR and varicella vaccines followed 6 weeks later by 2 doses of hepatitis A vaccine 6 to 9 months apart (n = 455). Immune responses were evaluated at baseline, 31 days after the second dose of hepatitis A vaccine, and 42 days after MMR and varicella vaccine administration. Solicited, unsolicited, and serious adverse events were collected. RESULTS: After 2 doses of hepatitis A vaccine, nearly all subjects in all groups were seropositive (>=99%). Coadministration of hepatitis A vaccine with MMR and varicella vaccines did not impact the immunogenicity of any of the vaccines and was well tolerated. CONCLUSIONS: The immune response to hepatitis A vaccine and US-licensed MMR and varicella vaccines is not adversely affected when coadministered in children 15 months of age. PMID- 21617574 TI - The use of blood counts and blood cultures to screen neonates born to partially treated group B Streptococcus-carrier mothers for early-onset sepsis: is it justified? AB - BACKGROUND: No clear recommendations exist regarding the approach to evaluate neonates born to partially treated group B Streptococcus (GBS)-carrier mothers for early-onset GBS (EO-GBS) sepsis. OBJECTIVE: To determine the yield and drawbacks of screening, all neonates born to GBS-carrier mothers who received only one dose of IV antibiotic, less than 4 hours, before delivery (partially treated). METHODS: A retrospective analysis was performed of all complete blood counts (CBCs) and blood cultures obtained from infants born during the period 2005 to 2009 to GBS-positive screened mothers treated with only one dose of antibiotic prior to delivery. A review was conducted of all neonatal EO-GBS sepsis cases during the study period. RESULTS: : Of 5845 GBS-carrier mothers, 1648 (28%) received only one dose of antibiotic less than 4 hours before delivery. We traced the CBCs and blood cultures, which were taken from 1413/1648 (86%) infants after birth. In 234 (18%) of these 1413 neonates, a second CBC sample was taken due to abnormal result of the CBC (leukocytosis, leukopenia, or thrombocytopenia) or secondary to technical failure in obtaining the blood. None of the blood cultures taken in that screening protocol was GBS positive, but in 10 cases contamination with coagulase-negative Staphylococcus was reported. During the study period, EO-GBS sepsis was diagnosed in 11 neonates; all had clinical symptoms upon presentation. CONCLUSIONS: The use of CBC and blood culture to screen neonates born to GBS-carrier mothers who received only one dose of IV antibiotic before delivery led to a negligible clinical yield and a high rate of technical failure. Although these findings are in line with the recent change in the Centers for Disease Control guidelines, they put in question the cost of this practice in terms of neonatal pain and parental anxiety. PMID- 21617575 TI - Peroxisome proliferator-activated receptor gamma agonist troglitazone inhibits high mobility group box 1 expression in endothelial cells via suppressing transcriptional activity of nuclear factor kappaB and activator protein 1. AB - High mobility group box 1 (HMGB1), a delayed mediator of proinflammatory cytokines, could initiate and amplify inflammatory responses to infection, injury, and other inflammatory stimuli, and it has emerged as a potential therapeutic target for inflammatory diseases. The overexpression of HMGB1 in endothelial cells has been proved to contribute to the development of these diseases. Because many proinflammatory cytokines expression were suppressed by thiazolidinediones (TZDs), agonists for nuclear receptor peroxisome proliferator activated receptor gamma (PPARgamma), whether TZDs can inhibit HMGB1 expression and function is of great interest, however, it remains unknown. Herein, we provide evidence that PPARgamma agonist troglitazone, a member of the TZD class, modulates HMGB1 expression in the endothelial cell line EA.hy926 and propose a potential mechanism for that. Results from polymerase chain reaction experiments revealed that PPARgamma is expressed in EA.hy926 cells, and it can be activated by troglitazone. Troglitazone inhibited the basal and LPS-stimulated HMGB1 expression at the mRNA level and protein level. A luciferase reporter assay showed that troglitazone inhibited not only the transcriptional activation of the HMGB1 promoter but also activities of heterologous promoters driven by nuclear factor kappaB (NF-kappaB) or activator protein 1 (AP-1) response elements. Altogether, these data suggest that NF-kappaB and AP-1 may participate in the inhibitory effect on HMGB1 transcription induced by troglitazone. Activation of PPARgamma by troglitazone is effective for HMGB1 inhibition via suppressing NF kappaB and AP-1 transcriptional activity in endothelial cells, which provides a new potential strategy to suppress excessive HMGB1 in inflammatory diseases. PMID- 21617576 TI - Modulation of innate immunity by adenosine receptor stimulation. AB - In the past decades, increased concentrations of the signaling molecule adenosine have been shown to play an important role in the prevention of tissue damage evoked by several stressful circumstances. During systemic inflammation, the circulating adenosine concentration increases rapidly, even up to 10-fold in septic shock patients. By binding to specific adenosine receptor subtypes, designated A1, A2a, A2b, and A3, adenosine exerts a wide variety of immunomodulating and (cyto)protective effects. Only recently, several specific adenosine receptor agonists and other drugs that modulate adenosine metabolism have been developed for human use. Importantly, correct interpretation of the effects of adenosine is highly related to the model of inflammation used, e.g., administration of endotoxin or live bacteria. This review will discuss the potential role for adenosine as an immunomodulating and cytoprotective signaling molecule and will discuss its potential role in the treatment of the patient suffering from sepsis. PMID- 21617577 TI - Early administration of sivelestat, the neutrophil elastase inhibitor, in adults for acute lung injury following gastric aspiration. AB - Gastric aspiration is the major cause of acute lung injury (ALI) and acute respiratory distress syndrome. Aspiration-induced ALI is believed to be, at least in part, facilitated by neutrophil-derived mediators and toxic molecules. We conducted a prospective cohort study based on the hypothesis that sivelestat, a specific neutrophil elastase inhibitor, is effective for treating ALI following gastric aspiration. Forty-four ALI patients who showed evidence of aspiration were observed within 12 h before intensive care unit admission and who had been mechanically ventilated within 12 h after admission were included in this study. Lung injury score (LIS) and PAO2/FiO2 (P/F) ratio on day 7 were defined as the primary outcomes of the study. Twenty-three patients were assigned to the sivelestat group and 21 to the control group. In univariate analyses, the proportions of patients with LIS lower than 1.0 on day 7 and a P/F greater than 300 on day 7 were significantly higher in the sivelestat group than in the control group (60.9% vs. 26.3%, P = 0.03; 87.0% vs. 36.8%, P = 0.001). In the logistic regression model, the use of sivelestat was an independent predictor for LIS lower than 1.0 on day 7 (relative risk, 7.4; 95% confidence interval [CI], 1.51-36.48) and for a P/F ratio higher than 300 on day 7 (relative risk, 18.5; 95% CI, 2.72-126.46). In the Cox proportional hazards model, the use of sivelestat was associated with a lower cumulative proportion of patients who received mechanical ventilation during the initial 14 days (hazard ratio, 2.6; 95% CI, 1.17-5.55). PMID- 21617578 TI - The liver as a central regulator of hydrogen sulfide. AB - The liver is likely exposed to high levels of hydrogen sulfide (H2S) from endogenous hepatic synthesis and exogenous sources from the gastrointestinal tract. Little is known about the consequence of H2S exposure on the liver or hepatic regulation of H2S levels. We hypothesized that the liver has a high capacity to metabolize H2S and that H2S oxidation is decreased during sepsis, a condition in which hepatic O2 is limited and H2S synthesis is increased. Using a nonrecirculating isolated and perfused liver system, we demonstrated rapid hepatic H2S metabolism up to an infusion concentration of 200' MUM H2S. Hydrogen sulfide metabolism was associated with an increase in O2 consumption from a baseline 96.7 +/- 7.6 MUmol O2/min/kg to 109 +/- 7.4 MUmol O2/min/kg at an infusion concentration of 150 MUM H2S (P < 0.001). Removal of O2 from the perfusate decreased H2S clearance from a maximal 97% to only 23%. Livers isolated from rats subjected to cecal ligation and puncture (CLP) did not differ significantly from control livers in their capacity to metabolize H2S, suggesting that H2S oxidation remains a priority during sepsis. To test whether H2S induces O2 consumption in vivo, intravital microscopy was utilized to monitor the oxygen content in the hepatic microenvironment. Infusion of H2S increased the NADH/NAD+ ratio (645 gray-scale-unit increase, P = 0.035) and decreased hepatic O2 availability visualized with Ru(Phen)3(2+) (439 gray-scale-unit increase, P = 0.040). We conclude that the liver has a high hepatic capacity for H2S metabolism. Moreover, H2S oxidation consumes available oxygen and may exacerbate the tissue hypoxia associated with sepsis. PMID- 21617579 TI - beta2-adrenergic receptor antagonist butoxamine partly abolishes the protection of 100% oxygen treatment against zymosan-induced generalized inflammation in mice. AB - We have demonstrated that 100% oxygen inhalation is beneficial to zymosan-induced generalized inflammation, and reactive oxygen species may be involved in the protection of oxygen treatment. Other investigators suggest that reactive oxygen species may modulate the sympathetic nervous system activity and beta2-adrenergic receptor (beta2AR)-mediated pathway. Moreover, studies have demonstrated that beta2AR agonists are beneficial to sepsis. Therefore, we assessed the effects of beta2AR antagonist butoxamine on the protection of oxygen treatment against zymosan-induced generalized inflammation in mice. Mice were given oxygen treatment by exposure to 100% oxygen for 3 h starting at 4 and 12 h after zymosan injection, respectively. In the mortality study, survival was monitored for 7 days after zymosan injection in mice. At 24 h after zymosan injection, mice were killed, and blood sample and organs were harvested for analysis. We observed that 100% oxygen treatment prevented the abnormal changes in organ histopathology, lactate dehydrogenase and C-reactive protein in serum, inflammatory cytokines in serum and tissue, and arterial blood gas analysis and improved the survival rate in zymosan-challenged mice. We found that pretreatment with beta2AR antagonist butoxamine partly abolished the protection of 100% oxygen inhalation. We also showed that zymosan induced the increase in serum 3'-5'-cyclic adenosine monophosphate (cAMP) and the decrease in tissue cAMP. However, oxygen treatment increased the cAMP levels in both serum and tissue, which were partly abolished by pretreatment with butoxamine. Thus, 100% oxygen inhalation may protect against zymosan-induced generalized inflammation in mice partly through activation of beta2AR pathway and subsequently enhance cAMP levels in both serum and tissue. PMID- 21617580 TI - Severe intestinal ischemia can trigger cardiovascular collapse and sudden death via a parasympathetic mechanism. AB - Hemorrhagic shock and splanchnic arterial occlusion (SAO) followed by reperfusion are associated with high mortality. However, rapid cardiovascular failure and death may also occur before reperfusion in hemorrhagic shock and SAO. We show in a rat SAO model that, upon gut ischemia, mean arterial blood pressure transiently elevates and then drops fatally in one of two time courses: (i) gradually over ~1 to 3 h or (ii) rapidly (often by >80 mmHg) over a period of 1 to 6 min. We hypothesize that fast fatal pressure drops (FFPDs) are due to failure of autonomic nervous system control. To test this, we treated rats with Glucose (10%) in the small intestinal lumen and intramuscularly administered xylazine to activate the parasympathetic nervous system or with a muscarinic anticholinergic (glycopyrrolate) or by total subdiaphragmatic vagotomy to attenuate parasympathetic nervous system activity. We also tested nafamostat mesilate (ANGD [6-amidino-2-naphthyl p-guanidinobenzoate dimethanesulfonate]), a protease inhibitor efficacious in preventing blood pressure loss in SAO with reperfusion, in the intestinal lumen. Fifty percent of animals receiving xylazine and Glucose died by FFPD (vs. 33% with neither, not statistically significant). Total subdiaphragmatic vagotomy or glycopyrrolate treatment significantly reduced the incidence to 0% (P < 0.008), although slow fatal pressure drops still occurred. ANGD did not prevent FFPDs, but delayed onset of slow fatal pressure drops (P < 0.013). These results suggest that gut ischemia can cause sudden death via an autonomic nervous system mechanism and that SAO with Glucose and xylazine may serve as a useful model for the study of neurogenic shock or autonomic dysregulation associated with sudden death. PMID- 21617582 TI - Accurate staging for gallbladder cancer: implications for surgical therapy and pathological assessment. AB - BACKGROUND: This study evaluates the significance of tumor involvement of the liver in early T-stage tumors and lymph node (LN) metastases on outcome after R0 resection of gallbladder cancer (GBCA). METHODS: A prospectively maintained database, supplemented with review of the medical record, was used to identify patients who underwent a complete (R0) resection for GBCA. All patients underwent definitive surgical treatment at the initial operation (1 stage) or after initial noncurative cholecystectomy (incidental tumors, 2 stage), including partial hepatectomy and portal LN dissection, with or without bile duct and/or adjacent organ resection. Clinicopathological variables, including TNM stage, histologic tumor involvement of liver (residual or direct extension in the GB fossa or discontiguous disease), and the total number of regional LNs assessed were analyzed for their association with outcome. RESULTS: One hundred twenty-two patients were identified and analyzed. The median follow up period was 23 months. Liver and nodal involvement by GBCA were observed in 61 (50%) and 41(34%) patients, respectively. Among patients with T2 tumors (n = 53), 48 (91%) were incidental. Liver involvement was present in 26%, and this factor was associated with decreased recurrence-free (RFS) and disease-specific survival (DSS) compared with patients with T2 tumors without liver involvement (median RFS, 12 months vs. not reached, P = 0.004, median DSS 25 months versus not reached, P = 0.003); T1b tumors (n = 10) were not associated with liver involvement. The median total lymph node count (TLNC) was 3 (range 0-20). For the entire cohort, survival of patients classified as N0 based on TLNC < 6 was significantly worse than that of N0 patients based on TLNC >= 6 (median RFS, 22 months versus not reached, P < 0.001, median DSS 41 months versus not reached, P < 0.001). Liver involvement and TLNC remained significant prognostic factors in a multivariate model that included TNM stage. CONCLUSION: Resection and histologic evaluation of at least 6 lymph nodes improves risk-stratification after resection of GBCA. Incidental T2 tumors are often associated with residual liver disease and should be reclassified to reflect the adverse outcome. The data suggests a need for standardized minimum requirements for adequate surgical treatment and pathological examination. PMID- 21617583 TI - Surgical cure for type II diabetes: myth or reality? PMID- 21617585 TI - The long-term impact of surgical complications after resection of stage I nonsmall cell lung cancer: a population-based survival analysis. AB - OBJECTIVE: Surgical morbidity may influence long-term cancer survival. Because resection of early stage nonsmall cell lung cancer (NSCLC) is primary therapy, we sought to determine the survival impact of surgical complications for elderly patients undergoing resection of stage I NSCLC. METHODS: Using the linked Surveillance Epidemiology and End Results-Medicare database (2000-2005), we identified elderly patients who underwent lobectomy for stage I NSCLC. We then assessed the unadjusted association between in-hospital, postoperative complications, and long-term survival for patients who survived more than 30 days after resection using the Kaplan-Meier method. Finally, we used Cox proportional hazards regression to evaluate the relationship between postoperative complications and 5-year cancer-specific (CSS) and overall survival (OS) after adjusting for patient, tumor, and treatment characteristics. RESULTS: We identified 3996 eligible patients. The overall in-hospital, postoperative complication rate was 54.2%. Pulmonary complications were the most common (n = 1464) followed by cardiac (n = 916). Unadjusted 5-year CSS was significantly worse for those who had an in-hospital, postoperative complication (70.9%) compared to those who did not (78.9%, P < 0.001). OS was also significantly worse (P < 0.001) for patients who developed a complication. Complications continued to predict worse 5-year CSS and OS after adjusting for patient, tumor, and treatment characteristics (HR: 1.38, 95% CI, 1.17-1.64). CONCLUSIONS: The occurrence of in hospital postoperative complications was an independent predictor of worse 5-year CSS after resection of stage I NSCLC. Importantly, the impact of surgical complications extends well after the initial perioperative period. These findings may help identify important targets for best practice guidelines and quality-of care measures. PMID- 21617586 TI - Early and late acute antibody-mediated rejection differ immunologically and in response to proteasome inhibition. AB - BACKGROUND: The efficacy of plasma cell targeted therapies for antibody-mediated rejection (AMR) has not been defined in detail. The purpose of this study was to compare early and late acute AMR in terms of immunologic characteristics and responses with proteasome inhibitor (PI) therapy. METHODS: Renal transplant recipients with acute AMR were treated with PI-based regimens. Early acute AMR was defined as occurring within 6 months posttransplant. Immunodominant donor specific antibody (iDSA) was defined as the DSA with the highest level. RESULTS: Results are expressed as early or late acute AMR. Thirty AMR episodes (13 early, 17 late) were treated in 12 and 16 patients. Early but not late AMR was associated with presensitization. Late AMR iDSA levels were higher, and specificities were primarily class II (DQ being most frequent). Early AMR patients demonstrated greater reduction in iDSA at 7, 14, and 30 days and at the posttreatment nadir (81.5%+21.2% vs. 51.4%+27.6%; P<0.01). Early AMR patients were more likely to demonstrate histologic resolution/improvement (87.5% vs. 53.8%; P=0.13). Both groups demonstrated significant improvement in renal function. CONCLUSIONS: Early and late AMR exhibit distinct immunologic characteristics and respond differently to PI therapy. PMID- 21617587 TI - Chronic kidney disease after orthotopic liver transplantation: impact of hepatitis C infection. AB - BACKGROUND: Hepatitis C virus (HCV) infection has been shown to be a potential risk factor for the development of chronic kidney disease in liver transplant recipients. METHODS: We performed a retrospective cohort study of 307 patients with and without HCV cirrhosis and preserved pretransplant renal function (serum creatinine<1.5 mg/dL pretransplantation) to assess the impact of HCV on the incidence of posttransplant chronic kidney disease. Kaplan-Meier analysis was performed for time to development of estimated glomerular filtration rate (eGFR) less than 30 mL/min, need for dialysis, and mortality. RESULTS: One hundred eighty-one patients were transplanted for HCV cirrhosis and 126 recipients had other causes of liver disease. Mean model for end-stage liver disease scores were 21.64 in the HCV group and 21.30 in the non-HCV group (P=0.58); 51% of patients in the HCV cohort had hepatocellular carcinoma compared with 27% in the non-HCV cohort (P<0.001). Mean pretransplant serum creatinine level was 0.89 mg/dL in both groups. At 3 years posttransplant, eGFR did not differ between the HCV and non-HCV cohorts (64.96 mL/min vs. 66.09 mL/min; P=0.71). A total of 14.4% of the patients with HCV achieved an eGFR less than 30 mL/min compared with 10.3% of the patients without HCV (P=0.13). There was no difference between the cohorts with respect to requirement for dialysis (P=0.73) or deaths (P=0.08), including those that were liver related (P=0.15). CONCLUSIONS: Patients with HCV cirrhosis and normal preliver transplant renal function do not have worse posttransplant renal outcomes compared with those with other causes of liver disease. PMID- 21617588 TI - Basiliximab induction and delayed calcineurin inhibitor initiation in liver transplant recipients with renal insufficiency. AB - BACKGROUND: Renal insufficiency (RI) is common after liver transplantation (LT) and may worsen due to calcineurin inhibitor (CNI) use. We compared LT outcomes using basiliximab induction and delayed CNI initiation to controls with a standard CNI regimen in patients with peri-LT RI. METHODS: All adults transplanted January 2004 to December 2007 with peri-LT RI (hemodialysis or creatinine >=1.5 within 1 week of LT) were included in a retrospective nonrandomized cohort. Outcomes including 30-day and 1-year patient and graft survival and renal function were compared between basiliximab and control groups. RESULTS: Two hundred twenty-nine patients (102 basiliximab, 127 controls) were analyzed, mean age 54 years, 72% men, 54% with hepatitis C virus. Mean model for end-stage liver disease (28.2 vs. 20.0; P<0.001) and creatinine (1.9 vs. 1.6; P=0.001) were higher and more patients were on hemodialysis at LT (29% vs. 6%; P<0.001) in the basiliximab group. 30-day patient (99% vs. 97%; P=0.26) and graft survival (98% vs. 95%; P=0.17), 1-year patient (87% vs. 87%; P=0.89) and graft survival (86% vs. 82%; P=0.37), mean creatinine at 1-year (1.5 vs. 1.5 mg/dL; P=0.82), and treated acute rejection (6% vs. 6%; P=0.90) were similar between basiliximab and control groups, respectively. In multivariable logistic regression, basiliximab was not significantly associated with 30-day (odds ratio, 0.10; P=0.11) or 1-year (odds ratio, 0.97; P=0.94) survival, controlling for age, previous LT, model for end-stage liver disease, and hepatitis C virus. CONCLUSIONS: Basiliximab induction resulted in 30-day and 1-year patient, graft and renal outcomes comparable with a control group receiving standard CNI-based immunosuppression. Antibody induction with delayed CNI should be further studied prospectively. PMID- 21617589 TI - Aprotinin versus tranexamic acid during liver transplantation: impact on blood product requirements and survival. AB - BACKGROUND: Historically, orthotopic liver transplantation (OLT) has been associated with major blood loss and the need for blood product transfusions. Activation of the fibrinolytic system can contribute significantly to bleeding. Prophylactic administration of antifibrinolytic agents was found to reduce blood loss. METHODS: The efficacy of two antifibrinolytic compounds--aprotinin (AP) and tranexamic acid (TA)--was compared in OLT. Four hundred consecutive OLTs were studied: 300 patients received AP and 100 received TA. Multivariate logistic regression analysis was used to identify independent predictors of intraoperative transfusion requirement and 1-year patient mortality. RESULTS: There was no intergroup difference in intraoperative blood loss (1082+/-1056 vs. 1007+/-790 mL), red blood cell transfusion per patient (0.5+/-1.4 vs. 0.5+/-1.0), final hemoglobin (Hb) concentration (93+/-20 g/L vs. 95+/-22 g/L), the percentage of OLT cases requiring no blood product administration (80% vs. 82%), and 1-year survival (85.1% vs. 87.4%). Serum creatinine concentrations were also the same (116+/-55 vs. 119+/-36 MUmol/L) 1 year after surgery. Two variables, starting Hb and phlebotomy, correlated with the two primary outcome measures (transfusion and 1-year survival). CONCLUSIONS: In our experience, administration of AP was not superior to TA with regards to blood loss and blood product transfusion requirement during OLT. In addition, we found no difference between the groups in the 1-year survival rate and renal function. Furthermore, we suggest that starting Hb concentration should be considered when prioritizing patients on the waiting list and planning perioperative care for OLT recipients. PMID- 21617590 TI - Analysis of factors affecting recurrence of hepatocellular carcinoma after liver transplantation with a special focus on inflammation markers. AB - BACKGROUND: Systemic inflammation markers, such as neutrophil-to-lymphocyte ratio (NLR), have recently emerged as the prognostic factors for recurrence of liver tumors. METHODS: We assessed the ability of NLR and of other variables to predict the outcomes of orthotopic liver transplantation (OLT) for hepatocellular carcinoma (HCC). A retrospective analysis was performed in 219 patients with HCC who underwent OLT between 1997 and 2009, with a median follow-up of 40 months. RESULTS: Overall 3- and 5-year patient survival rates were 76.6% and 70.7%, respectively. Overall 3- and 5-year recurrence-free survival (RFS) rates were 83.8% and 82.1%, respectively. On univariate analysis, the factors affecting overall survival were alpha-fetoprotein more than 30 ng/mL (P=0.006), NLR more than or equal to 5 (P<0.0001), hepatitis C infection (P=0.043), and presence of microvascular invasion (MVI; P=0.006). Preoperative treatments (P=0.006), alpha fetoprotein more than 30 ng/mL (P=0.003), NLR more than or equal to 5 (P<0.0001), exceeding Milan criteria at final histology (P=0.001), poor tumor differentiation (P=0.02), and presence of MVI (P<0.0001) predicted a lower RFS. Cox's proportional hazard model showed that only increased NLR and presence of MVI independently predicted overall survival and RFS. CONCLUSIONS: NLR is an important predictor of outcome after OLT for HCC and should be used to identify OLT candidates at high risk of recurrence. PMID- 21617591 TI - Platycoside O, a new triterpenoid saponin from the roots of Platycodon grandiflorum. AB - A new unusual minor triterpenoid saponin, platycoside O (1), was isolated from the 75% EtOH extract obtained from the roots of Platycodon grandiflorum, together with four known saponins: platycoside M-3 (2), platycoside J (3), platycoside F (4) and platycoside B (5). The structure of 1 was determined as 3-O-beta-D glucopyranosyl-(1->6)-beta-D-glucopyranosyl-2beta,3beta,16alpha,23 tetrahydroxyolean-12-en-24-methoxyl, 24-oxo-28-oic acid 28-O-beta-D-xylopyranosyl (1->4)-alpha-L-rhamnopyranosyl-(1->2)-alpha-L-arabinopyranoside on the basis of spectral analysis and chemical evidence. PMID- 21617592 TI - Dexmedetomidine (Precedex) for ICU sedation. AB - Opioids, benzodiazepines, propofol, antipsychotics and dexmedetomidine (Precedex) are frequently used in the intensive care unit (ICU) to manage pain, anxiety, agitation and delirium, and to facilitate procedures such as mechanical ventilation. The use of dexmedetomidine, a centrally-acting selective alpha(2) receptor agonist approved by the FDA in 1999, has been increasing in recent years. Some new studies comparing it to other drugs for ICU sedation have been published. PMID- 21617593 TI - In brief: fexofenadine (Allegra) and fruit juice. PMID- 21617594 TI - Capsaicin patch (Qutenza) for postherpetic neuralgia. AB - The FDA has approved a topical 8% patch formulation of capsaicin (Qutenza NeurogesX), available only by prescription, for local treatment of postherpetic neuralgia. Postherpetic neuralgia occurs after herpes zoster in about one third of patients >=60 years old and can persist for months or even years. PMID- 21617595 TI - New treatments for keratoconus. AB - Keratoconus is a fairly common, progressive, noninflammatory, corneal-thinning disorder in which the cornea has a conical shape. Rigid gas permeable (RGP) contact lenses or corneal transplantation are effective treatments, but intracorneal ring segments and collagen cross-linking procedures offer some new alternatives in the management of the disease. PMID- 21617596 TI - Drugs for HIV infection. PMID- 21617597 TI - Coxibs: is there a benefit when compared to traditional non-selective NSAIDs in postoperative pain management? AB - A multi-modal approach for the management of postoperative pain has become increasingly popular. Strategies to avoid the use of opioids and thus any opioid analgesic related side-effect is an important part of the expansion of ambulatory surgery. Combining long acting local anesthesia in the wound area and non-opioid analgesics are today a basic concept in management of day care, short stay patients. Paracetamol and non-steroidal anti-inflammatory drugs (NSAIDs) is often sufficient to provide satisfactory pain relief after minor and intermediate procedures. The use of multimodal or balanced analgesia has since long been shown to facilitate resumption of activities of daily living. The opioid sparing effects of the addition of NSAIDs to morphine patient-controlled analgesic (PCA) after major surgery has also shown repeatedly. The development and introduction of the most selective cyclo-oxygenase-2-inhibitors (Coxibs) was primarily indicated to reduce the risk and severity of gastrointestinal bleeding. The Coxibs have become an interesting option in postoperative pain management. The less pronounced effect on platelet function and subsequent lower risk for impaired hemeostasis makes them, in theory, a preferred option to the non selective traditional NSAIDs. The benefit versus risk for a more generalized use of Coxibs must, however, be based on a thorough evaluation of the overall benefits and risks for the use of NSAIDs and a further evaluation on whether the specific therapeutic features of the Coxibs provide benefits outweighing their increased cost. This review aims at providing a background and an overview of the benefits versus risks for the use of Coxibs as part of a multimodal postoperative pain management. PMID- 21617598 TI - Impact of early vs. late tracheostomy on weaning: a retrospective analysis. AB - BACKGROUND: Early tracheostomy has been advocated for a number of reasons. Especially in association with weaning from mechanical ventilation, it is known that an early timepoint can help patients being weaned more rapidly from the ventilator. However, timing of tracheostomy is still unknown and evidence is lacking. The effects of early tracheostomy compared with intermediate and late tracheostomy were assessed in critically ill patients. METHODS: Data collected from January 2005 to December 2007 were conducted for retrospective analysis. All patients needing tracheostomy due to extubation failure and/or weaning failure were included (N.=296). Early tracheostomy (ET) was defined as <=4 days, intermediate tracheostomy (IT) as tracheostomy within 5-9 days, and late tracheostomy (LT) was defined as >=10 days after endotracheal intubation. After proving normal distribution, significant changes between the three groups were tested by ANOVA followed by post hoc tests for multiple comparisons (Bonferroni's test). RESULTS: Intensive care unit (ICU) mortality was significantly higher in the LT group when being compared with the ET but not when being compared with the IT group (40.7% vs. 24.8% vs. 17.1%). Further, a significantly reduced incidence of VAP and sepsis, a smaller amount of ventilator days and a shorter ICU length of stay could be observed for the ET group. Length of weaning was not significantly different between the groups. CONCLUSION: The length of weaning after tracheostomy is not affected by the timing. It seems beneficial to favour early tracheostomy in order to reduce the time of mechanical ventilation and its associated risks. PMID- 21617599 TI - Usefulness and limits of near infrared spectroscopy monitoring during endovascular neuroradiologic procedures. AB - BACKGROUND: Transcranial cerebral oximetry (TCCO) with near-infrared spectroscopy (NIRS) is a non-invasive, bedside technique, which allows the continuous measurement of regional cerebral oxygenation. The aim of this study was to evaluate TCCO monitoring during endovascular neuroradiologic procedures. METHODS: Adult patients undergoing elective endovascular embolization of cerebral aneurysms, arteriovenous malformations, dural arteriovenous fistulas and meningiomas under general anesthesia were included in the study, over a period of 12 months. Twenty-eight procedures in 25 patients were analyzed. RESULTS: Regional cerebral oxygenation rSO(2) readings were significantly different according to the different phases of the neuroendovascular procedure. An effect of the underlying cerebral pathology on regional cerebral oxygenation rSO(2) recording, in relation to the different stage of the interventional procedure, was also evident, the more invasive the procedure the greater the impact on rSO(2) reading. NIRS monitoring contributed to a prompt diagnosis and management of two adverse intraoperative events and helped in early evaluation of prognosis. CONCLUSION: TCCO with NIRS is a promising monitoring tool to assess the balance between oxygen supply and demand during neuroradiologic procedures. Nevertheless, some limits should be acknowledged, such as the study of the posterior circulation and artefacts related to contrast agent injection. A careful understanding of the undergoing step of the procedure as well of the possible influence of intrinsic and extrinsic factors affecting recording is important for interpretation of data. PMID- 21617600 TI - Long-term extracorporeal membrane oxygenation with minimal ventilatory support: a new paradigm for severe ARDS? AB - Pulmonary tuberculosis can lead to acute respiratory distress syndrome (ARDS) which is associated with high mortality. We report the case of a patient with pulmonary tuberculosis and severe ARDS (PaO2/FiO2<100 mmHg) who was initially managed with advanced up-to-date treatments (protective ventilation and extracorporeal membrane oxygenation, ECMO) but failed to improve. After a month of failure and the development of bilateral pneumothoraces, we drastically changed our therapeutic strategy: we maximized ECMO support to maintain oxygenation, we greatly reduced ventilation pressures and we left the pneumothoraces undrained. From then on, the patient improved and he eventually survived. This case suggests that ECMO permits large reductions in lung inflation and ventilation to rest the lungs, while maintaining acceptable oxygenation. The combination of ECMO and markedly attenuated ventilation strategy may be effective in cases of severe ARDS. PMID- 21617601 TI - The 2011 revolution in antimicrobial susceptibility testing and reporting: what the ICU physicians need to know. PMID- 21617602 TI - Pediatric delirium in the pediatric intensive care unit: a systematic review and an update on key issues and research questions. AB - There was a dearth in awareness and knowledge regarding pediatric delirium (PD) at the pediatric intensive care unit (PICU). The aim of this study is to highlight the most recent and up-to-date findings of current literature -by means of a systematic review (SR) method-, and to present the key issues and research questions. A SR of the literature published between 24 March 2009 and 10 March 2011. Eight new articles were identified which included (literature) reviews, journal commentaries, and observational studies. There still is a dearth of literature on PD, with also a variable level of evidence (1b - 5), especially in relation to critical illness. To be able to further clarify PD in the PICU, additional research questions are provided for future research. PMID- 21617603 TI - Prophylaxis of venous thromboembolism: low molecular weight heparin compared to the selective anticoagulants rivaroxaban, dabigatran and fondaparinux. AB - Newer therapeutic options available in the prevention of postoperative thromboembolism, currently focused on fondaparinux, rivaroxaban and dabigatran warrant an overall therapeutic assessment. The constitutive comparisons with enoxaparin are based on a combined outcome measure solely driven by the incidence of "asymptomatic deep vein thrombosis". Its validity as a clinically relevant endpoint is missing if antithrombotics of different classes are compared. This is because they target different phases of thrombogenesis i. e. ahead and beyond the asymptomatic stage of thrombosis. Additional concerns refer to the dosing regimens and their practical administration: Fondaparinux, rivaroxaban and dabigatran are dosed to achieve maximum effects very close to their limits of tolerance whereas wide dosing spectra for the low molecular weight herparin (LMWH)'s indicate the potential for dose adaptation and increase. The other disadvantage to the control-heparin originates in the timing for the 1st administration which doesn't fit in with the "just-in-time" principle. So the enoxaparin-regimen is lacking in benchmark-quality - with the consequence that the meaning of the Phase III-trials does'nt go beyond a mere technical demarcation from the marketed variant of the product as defined by the stipulations in the package insert. As to tolerance the selective anticoagulants exhibit an increased risk of major and other clinically relevant bleeding, exceeding that of enoxaparin by 30% (P<0.001). The outcome of the meta-analyses on fondaparinux, rivaroxaban and dabigatran is supported by product-specific calculations and assessments of the European Medicine Equivalence Agency (EMEA). Rivaroxaban and dabigatran show significant age-dependent renal accumulation. Because the dose-finding studies were restricted to patients over 60 year old the regimens definitely established are not applicable to younger patients. The reason for the limited therapeutic index of the selective anticoagulants originates in their monovalent activity as such not adequately matching the complexity of thrombogenesis and early thrombus extension. Their class-specific limitations are compensated through more intensive dosage-regimens which result in accentuated bleeding complications. Connotatively the hypothesis emerged that antiXa- and IIa-effects interact synergistically which translates into enhanced efficacy and tolerance. Experimental studies on hirudin with pentasaccharide and hirudin with "lower low molecular weight heparin" (3KDA) support such rationale. PMID- 21617604 TI - Distribution and clinical impact of phlebosclerosis. AB - Sclerosis of the vessels has been extensively described in arteries but rarely in veins. Phlebosclerosis refers to the thickening and hardening of the venous wall. Despite its morphological similarities with arteriosclerosis and potential morbid consequences phlebosclerosis has gained only little attention. This is a narrative review based on a MEDLINE search (1949 to February 2009) of articles in English, German and Japanese (with English abstracts). References in retrieved articles were searched for further potentially relevant articles. Controversy about the definition, etiology, pathogenesis and histological aspects of phlebosclerosis exists. Phlebosclerosis constitutes a fibrous degeneration of the venous wall, predominantly the intima, with or without calcification. It has been described to affect almost the entire venous system with variable clinical consequences. The exact pathophysiologic mechanisms are unknown. Despite the confusing terminology and limited scientific data, we conclude that phlebosclerosis should be regarded as a distinct clinical entity. Its prevalence and clinical significance may be underestimated. Future research towards the better definition of the term and recognition of its true prevalence, distribution and clinical significance is warranted. PMID- 21617605 TI - The association between subclinical atherosclerosis in carotid arteries and Connexin 37 gene polymorphism (1019C>T; Pro319Ser) in women. AB - AIM: The aim of our study was to evaluate a possible association between subclinical atherosclerosis in carotid arteries and the connexin 37 gene polymorphism (1019C>T; Pro319Ser) in a population of urban and rural women. METHODS: A 5% population sample of urban women aged 45-54 years (N.=896) and a 1% representative sample of rural women aged 33-72 years (N.=152) were examined using an identical protocol and genotyped for Cx37 gene polymorphism. The association between the Cx37 polymorphism and intima-media thickness in common carotid arteries measured by ultrasound (CIMT) was studied. RESULTS: We have found a different pattern of the effect of the Cx37 gene on CIMT with regard to fasting glycemia with significant interaction between fasting glycemia and Cx37 gene on CIMT (test for equality of slopes P<0.0001). In addition, we also detected potential threshold effect of fasting glycemia at the concentration of 5.5 mmol/L (ANCOVA; P=0.026). Carriers of TT genotype showed protection against subclinical atherosclerosis if their fasting glycemia was above 5.5 mmol/L. CONCLUSION: In women with higher fasting glycemia TT genotype of Cx37 polymorphism was protective against subclinical atherosclerosis. Therefore, the Cx37 gene may exert completely different effects in the artery wall, depending on glycemia. PMID- 21617606 TI - CALSFOAM-completed automated local statistics based first order absolute moment" for carotid wall recognition, segmentation and IMT measurement: validation and benchmarking on a 300 patient database. AB - AIM: In this work we present a novel methodology (called CALSFOAM) for the automated segmentation of ultrasound carotid images and intima-media thickness (IMT) measurement. CALSFOAM was developed in order to overcome limitations of a previously developed snake-based technique. METHODS: CALSFOAM consists of two stages: Stage-I is an automatic recognition of the carotid artery system in an image frame and Stage-II is a combination of segmentation and IMT measurement sub system. Stage-I is performed by using local statistics and by automatically tracing the profile of the distal adventitia. Stage-II takes the traced adventitia boundary and builds an ROI for distal wall segmentation that uses a first order absolute moment (FOAM) technique. CALSFOAM was benchmarked against our previous snake based technique and validated on a 300-image multi institutional dataset. RESULTS: CALSFOAM's lumen-intima (LI) segmentation error was 0.049+/-0.039 mm, the media-adventitia (MA) error was 0.088+/-0.054 mm; the IMT measurement bias was 0.125+/-0.103 mm. To reduce CALSFOAM error, we adopted a GREEDY approach for fusing the boundaries from the two techniques and obtained LI and MA errors equal to 0.02+/-0.014 mm, 0.023+/-0.013 mm, and an IMT bias of 0.074+/-0.068 mm. CONCLUSION: Even though CALSFOAM's performance was lower than snake-based segmentation techniques, it helped in avoiding possible inaccuracies of snakes and its parameter sensitivities. The very accurate performance obtained by the GREEDY approach demonstrated that the two techniques could be considered as complementary. PMID- 21617607 TI - Cardiac risk stratification in elective non-cardiac surgery: role of NT-proBNP. AB - AIM: The aim of the study was to investigate the utility of NT-proBNP measurement for the stratification of presurgical cardiac risk. METHODS: Cardiac risk before elective non-cardiac surgery was evaluated in 82 consecutive patients. From each patient a venous blood sample was drawn to determinate NT-proBNP levels. Patients were followed up over three months in order to detect the occurrence of cardiac adverse events. RESULTS: NT-proBNP was positively correlated (P<0.0001) with age, days of hospitalization (P=0.001) and ASA class (P=0.001). High surgical risk (P<0.0001), diabetes (P=0.004), dyslipidemia (P=0.006) and elevated levels of NT proBNP (P<0.0001) were significantly correlated with events. Using a logistic regression analysis we found an independent association between pre-operative elevated NT-proBNP and postoperative cardiac events (OR 1.2, 95% CI 1.0-1.4, P=0.01). CONCLUSION: Measuring NT-proBNP before non cardiac surgery in clinical practice could be useful to better stratify patients' risk. PMID- 21617608 TI - Effects of the ABO-mismatch between donor and recipient of cryopreserved arterial homografts. AB - AIM: Cryopreserved arterial homograft (CAH) is a well-established substitute material for in situ reconstruction of vascular infections. However, their degeneration remains serious complication. Although several studies propose ABO mismatching between CAH-donor and -recipient as the main reason, the results are controversial. We compared the outcome between ABO-compatible and ABO incompatible CAH recipients to evaluate the contribution of ABO-mismatching. METHODS: Between January 2004 and December 2007, a retrospective review in 32 patients who underwent CAH-implantation was performed. The patients were divided in ABO-incompatible (group A: 17/32 patients; 53%) and ABO-compatible (group B: 15/32 patients; 47%) to CAH donor. Leucocytes, platelets and C-reactive protein (CRP) levels were recorded during the in-hospital stay. These were correlated with the surface of implanted homograft (SIH). Mid-term survival- and freedom from-reoperation (FFR) rates were also calculated. RESULTS: In both groups, peak of leucocytes and CRP was recorded on third postoperative day (POD3) and regarding platelets lowest values on POD1. Interestingly, a second CRP-peak was reported on POD8 in group A (A: 172+/-104mg/L vs. B: 75+/-55mg/L, P=0.01). No relationship between second CRP-peak and SIH was found. After 27 months median follow-up (range, 5-49 months), survival- (65% vs. 84%, P=0.28) and FFR-rates (94% vs. 93%, P=0.98) remained comparable. CONCLUSION: We consider that the second CRP-peak expresses an early cytoimmunologic response of ABO-incompatible recipients against CAH. However, we did not find any relationship between ABO incompatibility and poor mid-term outcome in terms of reoperation or mortality. Longer surveillance of our patients is mandatory. PMID- 21617609 TI - A novel oscillometric device for peripheral arterial disease screening in everyday practice. The Czech-post MONICA study. AB - AIM: Ankle brachial index (ABI) is a diagnostic tool for peripheral arterial disease (PAD) and a cardiovascular risk stratification tool. Despite this evidence and guidelines recommending its use in everyday practice, ABI is not widely used. Automatic ABI measurement may lower the barrier to incorporate ABI measurement into everyday practice. The aim of this study was to validate a novel automatic oscillometric ABI device (BOSO ABI) against a gold standard-Doppler device in an epidemiological setting. METHODS: In 839 patients from the Czech post-MONICA study (a randomly selected representative population sample aged over 25 years), mean age 54.3+/-13.8 years (47% of men), ABI measurement was performed using the BOSO ABI device and a handheld Doppler device in a random fashion. The two techniques were carried out by different investigators each blinded to the findings of the other. Analyses were conducted as proposed by Bland and Altman. RESULTS: The mean ABI difference between the two methods was 0.1+/-0.11, with 95% limits of agreement ranging from -0.11 to 0.30. The difference between Doppler and oscillometric ABI increased significantly with increasing mean ABI (r=0.29; P<0.001). When considering Doppler the gold standard, automated oscillometric measurement had a 76.9% sensitivity, 97.9% specificity, and 37% positive and 99.6% negative predictive values in diagnosing ABI <0.9. CONCLUSION: The BOSO ABI device cannot be used interchangeably for standard Doppler ABI measurement in diagnosing PAD. However, its high negative predictive value allows using it as a screening tool for PAD. PMID- 21617610 TI - alpha-smooth muscle actin, fibrillin-1, apoptosis and proliferation detection in primary varicose lower limb veins of women. AB - AIM: The role of SMC apoptosis and proliferation was correlated to the amount of fibrillin and alfa-smooth muscle actin of primary varicose veins. METHODS: Twenty varicose vein specimens were atraumatically harvested from 20 women undergoing lower extremity primary varicose vein excision. The patients were divided into groups according to age (<50 years, >50 years) and the presence of leg edema (CEAP, class 2 or 3). The surface density of fibrillin-1 fibers (Sv([Fbn-1])), the volume density of smooth muscle cells: (Vv([SMC])), the number of proliferating and apoptotic cells per area. Quantitative data comparisons between class and age groups were performed. RESULTS: The median value of Vv([SMC]) was 16% greater and the Sv([Fbn-1]) was 35% greater in the intima vein sections from patients up to 50y compared to >50y. Apoptosis was found more frequent in veins sections from varicose women >50y. In the media layer, Sv([Fbn-1]) in veins from patients up to 50y was more important, and women with >50y had also more cells in apoptosis. Vv([SMC]) from women without edema (CEAP-Class 2) was 28% greater in the intima and apoptotic cells were more prominent in the intima of women with edema (CEAP-Class 2). In the media layer, Sv([Fbn-1]) was 12,5% greater in veins from women without edema and apoptosis was more detected in the veins from patients with edema. CONCLUSION: Age of the patient may affect the remodeling of varicose veins and SMC quantity in the media layer was found decreased in patients with edema. PMID- 21617611 TI - Correlation between improvement in functional signs and plethysmographic parameters during venoactive treatment (Cyclo 3 Fort). AB - AIM: The aim of this study was to compare the change in functional signs of venous insufficiency and venous refilling time measured by mercury strain-gauge plethysmography under the effects of the combination of Ruscus aculeatus, hesperidin methylchalcone and ascorbic acid (Cyclo 3 Fort). METHODS: An open label clinical trial conducted in 65 women presenting with CEAP class C2s and C3s assessment criteria. At D0 and D28, functional signs were measured on a visual analog scale (VAS), venous refilling time by mercury strain-gauge plethysmography and venous reflux by echo-Doppler method. RESULTS: Under treatment, venous refilling time increased from 11.7+/-4 s to 13.8+/-4.4 s with Delta of 26% (P<0.0001) and the percentage of patients showing improvement of functional symptoms >=30% was significantly correlated (P=0.04) with the percentage of patients presenting an improvement in venous refilling time >=10%. CONCLUSION: Improvement in subjective functional signs under treatment with Cyclo 3 Fort was correlated with objective plethysmographic parameter improvement. There-fore, this study contributes to objectively document the benefit of prescribing this venoactive drug to active women with CEAP class C2s to C3s. PMID- 21617612 TI - Magnetic resonance venography evaluating veins flow for legs by application of long stretch elastic bandage. AB - AIM: Varicose veins are a common feature of chronic venous disease that causes pain and swelling of legs. Long stretch elastic bandages are one of the methods used to prevent and treat venous disease, but diagnostic benefit by magnetic resonance imaging (MRI) is not widely promoted. This study aims to evaluate the effects of long stretch elastic bandages using a non-contrast magnetic resonance venographic (MRV) technique by detecting influence on circumferences and flow velocities of varicose and deep veins, before and immediately after application of long stretch elastic bandages. METHODS: Twelve patients presenting with visible varicose veins underwent MRV. The level of the superficial varicose and deep veins before and after wearing long stretch elastic bandages was recorded. RESULTS: By comparing before and after long stretch elastic bandages, all 12 cases showed statistical significant decrease of the superficial venous flow areas; 9 out of 12 cases showed statistical significant change of the superficial (decreased) and deep (increased) venous flow rates. However, due to technical limitation, over half of the cases could not record the change of the deep venous flow areas. CONCLUSION: The non-contrast MRV technique with Cine phase contrast, is capable to detect the short term beneficial effects of long stretch elastic bandages by measuring change of the superficial venous flow areas and superficial and deep venous flow rates. PMID- 21617613 TI - High bifurcation of the brachial artery. AB - Variations of the arterial pattern in the upper limb are quite common in humans. In the present study, we describe two cases of high bifurcation of the brachial artery. In the first cadaver both branches run superficially along the arm and were also located superficially at the antecubita fossa. In the second cadaver the branch that continues as the radial artery in the forearm run superficially and was located also superficially at the antecubita fossa, while the second branch had a more or less normal distribution. The embryological basis and clinical significance are discussed. PMID- 21617614 TI - Endovascular management of aortoenteric fistulas with aortic cuff extenders: report of two cases. AB - Despite immediate open surgery, aortoenteric fistula (AEF) remains a highly lethal condition. Endovascular management is widely employed, although there is no agreement on its role as a definite treatment or, because of a high incidence of recurrent bleeding and sepsis, as a bridge to open repair. Two cases of secondary AEFs after distant elective abdominal aortic aneurysm repair are presented. The first patient was a 76-year-old man and the second one a 70-year old man. Both patients presented with hematemesis, had no signs of sepsis and were successfully managed with endovascular surgery, using aortic cuff extenders. Postoperative course was uneventful for both patients who were discharged on long term antibiotics. However, during follow-up the first patient was readmitted four times; twice due to infection (at 2 and 6 months, respectively) and twice due to recurrent bleeding (at 5 and 9 months, respectively). The last episode of bleeding was managed with axillobifemoral bypass grafting, removal of the prostheses and closure of the aortic stump and the duodenal defect, but the patient died on the 5th postoperative day from multiple organ failure. The second patient remained asymptomatic until the 16th postoperative month when he developed lumbar spine osteomyelitis as a direct extension of graft infection and was deemed inoperable due to multiple comorbidities. Endovascular management of AEF can achieve satisfactory short-term results. Due to the high rate of recurrent bleeding and sepsis it should be used as a temporary measure and a bridge to open repair, whenever this is feasible. PMID- 21617615 TI - Efficacy and safety of long-term ezetimibe/simvastatin treatment in patients with familial hypercholesterolemia. PMID- 21617617 TI - Trauma-related death in the elderly: a new challenge for trauma system? PMID- 21617618 TI - Sleep in the ICU: where dreams go to die. PMID- 21617619 TI - Xenon exposure in the neonatal rat brain: effects on genes that regulate apoptosis. AB - BACKGROUND: In the developing rodent brain, exposure to volatile anesthetics causes widespread neuronal apoptosis in several regions of the brain. Increasing evidence points to a possible neuroprotective role for the anesthetic gas xenon, following neuronal injury. To address this gap in understanding, we explored the transcriptional consequences of xenon in the brains of postnatal day 7 (P7) rats exposed to xenon compared to those of air-breathing animals, with particular emphasis on the mRNA transcript levels of Akt and c-Jun N-terminal kinase kinase 1 (JNKK1), which are important for cell survival and the activation of extrinsic neuroapoptotic pathways, respectively. METHODS: P7 Sprague/Dawley rats were exposed to air (75% nitrogen, 25% oxygen) or xenon (75% xenon, 25% oxygen) for 120 min (N=6/group). Forebrains were harvested for reverse transcription polymerase chain reaction, which enabled quantification of Akt and JNKK1 mRNA transcripts. Suppression subtractive hybridization was used to explore the "genetic signature" of xenon exposure. RESULTS: Compared to control air-breathing animals, xenon-breathing rats exhibited a 0.7-fold decrease in Akt mRNA expression (P<0.01) and a 1.6-fold increase in JNKK1 mRNA levels (P<0.05). CONCLUSION: The concomitant decrease in the Akt mRNA expression level and increase in the JNKK1 mRNA transcript level provide evidence that xenon has a neuroapoptotic effect in the developing rodent forebrain. Given these results, further study into the paradoxical neuroprotective and neuroapoptotic effects of xenon is warranted. PMID- 21617620 TI - Contribution of the coronary sinus blood to the pulmonary artery oxygen saturation gradient in cardiac surgery patients. AB - BACKGROUND: The coronary sinus oxygen saturation (SO2) can affect the oxygen saturation of the superior vena cava (superior cava SO2) and the pulmonary artery (pulmonary artery SO2), causing a gradient between the latter two (DeltaSO2), as has been observed in different physiological and pathological conditions. The objective of the study was to evaluate the different determinants of DeltaSO2 in cardiac surgery patients. METHODS: An observational, prospective study was carried out on 18 patients undergoing elective cardiac surgery. Blood samples were obtained from the superior vena cava, the pulmonary artery, the inferior vena cava, and the coronary sinus before extracorporeal circulation. RESULTS: The following measurements were made: superior cava SO2, pulmonary artery SO2, coronary sinus SO2, and inferior cava SO2. The mean values (+/- SD) were as follows: superior cava SO2=76.4+/-12.6%; inferior cava SO2=72.7+/-15.8%; coronary sinus SO2=46.6+/-17.0%; and pulmonary artery SO2=71.9+/-12.9%. The DeltaSO2 was 4.5+/-5.5%. The average oxygen saturation (SO2avg=[Superior cava SO2 + inferior cava SO2]/2) was 74.6+/-13.7%. The superior cava SO2 was significantly higher than the pulmonary artery SO2, and the DeltaSO2 was significantly different from zero (P<=0.05). No significant differences were found between the superior cava SO2 and the inferior cava SO2, and both were significantly different from the coronary sinus SO2. The difference between SO2avg and the pulmonary artery SO2 was 2.74+/-4.4%. CONCLUSION: The observed DeltaSO2 could only be explained by dilution of the superior cava SO2 with blood with a lower SO2. The coronary sinus blood contributed to generate this gradient. PMID- 21617621 TI - Propofol, remifentanil and mivacurium: fast track surgery with poor intubating conditions. AB - BACKGROUND: Mivacurium is widespread used because it is the non-depolarizing muscle relaxant with the shortest duration time. Therefore, it seems to be ideal for fast track or ambulatory surgery. However, especially in combination with propofol and remifentanil onset time remains unclear and incidence of poor intubating conditions seems to be higher than in other regimes of anesthesia. METHODS: We included 35 ear, nose and throat (ENT) patients in this study. Muscle relaxation was measured by acceleromyograhpy at the adductor pollicis muscle (a.p.m.) and intubating conditions were evaluated. Anesthesia was induced with 2.5 mg kg-1 propofol and 1 ug kg-1 remifentanil and intubation was performed three minutes after the administration of 0.2 mg kg-1 mivacurium. Open vocal cords conjoined with full relaxation of the a.p.m., easy mouth opening and prevention of coughing and bucking represented the primary endpoint in this study. RESULTS: Only 20% of patients (N.=7) had optimal intubating conditions and achieved the primary endpoint. In 21 patients (60%) a complete block of the a.p.m. could not be achieved and in six patients (17%) the vocal cords were closed. In seven patients (20%) we observed difficult mouth opening and in 11 patients (31%) coughing and bucking. In addition, we found a prolonged onset time of 228+/-95 seconds (mean+/-SD). CONCLUSION: In combination with propofol and remifentanil the muscle relaxant agent mivacurium led to uncertain muscle relaxation and to poor intubating conditions. Therefore the study was aborted after 35 patients. Probably mivacurium is not a useful muscle relaxant agent if fast and deep muscle relaxation is needed. The advantage of a short duration time is foiled by intubation complications due to insufficient muscle relaxation. PMID- 21617622 TI - Age-related differences in mechanism, cause, and location of trauma deaths. AB - BACKGROUND: Trauma death has traditionally been described as primarily occurring in young men exposed to penetrating trauma or road traffic accidents. The epidemiology of trauma fatalities in Europe may change as a result of the increasing proportion of elderly patients. The goal of this study was to describe age-related differences in trauma type, mechanism, cause and location of death in a well-defined European region. METHODS: We prospectively registered all trauma patients and severe burn patients in eastern Denmark over 12 consecutive months. We analyzed all trauma fatalities in our region regarding the trauma type, mechanism, cause and location of death. RESULTS: A total of 2,923 patients were registered, of which 292 (9.9%) died within 30 days. Mortality increased with age, with a mortality of 46.1% in patients older than 80 years old. Blunt trauma was the most frequent trauma type at all ages, but the trauma mechanism differed among ages, with falls constituting 46.8% of trauma deaths in the elderly. The primary cause of death was head and spine injuries across all age-groups. Death took place before arrival at the hospital in 45% of the cases, but death during primary admission became increasingly important with advanced age. CONCLUSION: Increasing age was associated with higher mortality, an increased proportion of falls and fatal head or spine injuries. PMID- 21617623 TI - A recruitment maneuver improves apneic oxygenation. AB - BACKGROUND: Apneic oxygenation (AO) is applied during surgery and in intensive care units. Even with AO, apnea is associated with progressive hypoxemia, limiting the tolerable amount of time in AO. This experimental study evaluates the effects of a recruitment maneuver (RM) on oxygenation, CO2 retention, and survival times during prolonged apnea, supported or not supported with intratracheal apneic oxygenation. METHODS: Following Ethic Committee approval, 15 male Sprague-Dawley rats were anesthetized and ventilated with PCV and FiO2:1 for 15 minutes. After obtaining a basal arterial blood-gas sample, the rats were randomized into 3 groups and disconnected from the ventilator: group (G) 1 (N.=6): AO with a cannula inserted into the carina; G2 (N=6): RM (40 cm H2O CPAP applied for 30 seconds) before AO; and G3 (N.=3): no application after disconnection (G3 was stopped after the first 3 subjects died within 3 minutes). Further arterial blood-gas samples were taken after 1, 3, and 6 minutes (T1, T3, and T6). Survival times after the start of AO were recorded. RESULTS: G2 was associated with significantly higher values of PaO2 at T3 and T6 when compared to G1 (345+/-56 vs. 233+/-65 mm Hg at T3 and 258+/-31 vs. 180+/-31 mm Hg at T6, respectively, P<0.05). There were significant changes in PaO2, pH, and PaCO2 over time in all subjects, but no differences were observed between G1 and G2 in pH or PaCO2. Survival time in G2 was significantly longer as opposed to G1 (G1: 10.3+/ 2.3 min; G2: 14.3+/-3.6 min; P<0.05). CONCLUSION: RM prior to AO prolongs tolerance to apnea, probably by increasing the time before intolerable hypoxemia occurs, without a significant difference in PaCO2 levels. PMID- 21617624 TI - Sleep disturbances in the critically ill patients: role of delirium and sedative agents. AB - BACKGROUND: Impairment of sleep quality and quantity has been described in critically ill patients. Delirium, an organ dysfunction that affects outcome of the critically ill patients, is characterized by an acute onset of impaired cognitive function, visual hallucinations, delusions, and illusions. These symptoms resemble the hypnagogic hallucinations and wakeful dreams seen in patients with neurological degenerative disorders and suffering of disorders of rapid eye movement (REM) sleep. We assessed the characteristics of sleep disruption in a cohort of surgical critically ill patients examining the hypothesis that severe impairments of rapid eyes movement (REM) sleep are associated to delirium. METHODS: Surgical patients admitted to the intensive care units of the San G. Battista Hospital (University of Turin) were enrolled. Once weaning was initiated, sleep was recorded for one night utilizing standard polysomnography. Clinical status, laboratory data on admission, co-morbidities and duration of mechanical ventilation were recorded. Patients were a priori classified as having a "severe REM reduction" or "REM reduction" if REM was higher or lower than 6% of the total sleep time (TST), respectively. Occurrence of delirium during intensive care unit (ICU) stay was identified by CAM-ICU twice a day. Multivariate forward stepwise logistic regression analysis was performed with sleep ("severe REM reduction" vs. "REM reduction") as the a priori dependent factor. RESULTS: REM sleep amounted to 44 (16-72) minutes [11 (8-55) % of the TST] in 14 patients ("REM reduction") and to 2.5 (0-36) minutes [1 (0-6) % of the TST] in the remaining 15 patients ("severe REM reduction") (P = 0.0004). SAPS II on admission was higher in " severely REM deprived" then in "REM deprived" patients. Delirium was present in 11 patients (73.3%) of the patients with "severe REM reduction" and lasted for a median of 3 (0-11) days before sleep assessment, while only one patient having "REM reduction" developed delirium that lasted for 1 day. The factors independently associated with a higher risk of developing "severe REM reduction" were delirium and daily dosage of lorazepam. CONCLUSION: The present study shows that while all critically ill patients present a profound fragmentation of sleep with a high frequency of arousals and awakenings and a reduction of REM sleep, a percentage of patients present an extremely severe reduction of REM sleep. Delirium and daily dosage of lorazepam are the factors independently associated to extremely severe REM sleep reduction. PMID- 21617625 TI - Non-heart-beating organ donation in Italy. AB - In 2007 the non-heart-beating organ donation (NHBD) "Programma Alba" (Sunrise Programme) started in Pavia, Italy. The initial plan was to cut down waiting list for kidney transplantation, while its final aim is to shorten organ transplantation waiting lists. When compared to European countries and the USA, the Italian NHBD program has taken longer to get established. Initially Italian physicians were not entirely aware of the NHBD organ viability for transplantation, furthermore ethical issues and the need to regulate medical requirements to Italian law slowed down the NHBD program. In particular, Italian legislation provides for death ascertainment after irreversible cardiac arrest, 20-minute flat electrocardiogram. This no-touch period is longer when compared to worldwide legislation, and organ viability has been a main concern for Italian transplant doctors over the years. However, recent data let up to 40-minute warm ischemia time to preserve organ viability; this has encouraged Pavia's group to establish the NHBD "Programma Alba". It was designed according to Italian legislation from death diagnosis to graft placement, from this perspective must the significant role of the Transplant coordinator be recognized. Since 2007 seven kidneys have been gathered from seven NHBD. Of these, six NHBD kidneys have been transplanted. Currently, four patients are out of dialysis. This report is a detailed description of NHBD "Programma Alba" and its preliminary results. PMID- 21617626 TI - Pharmacogenetic treatment of acute respiratory distress syndrome. AB - Acute lung injury (ALI) and the acute respiratory distress syndrome (ARDS) occur due to systemic inflammatory disorders or direct injury to the lung. The occurrence of ALI/ARDS is sporadic and is not reliably predicted by the type or severity of injury. A combination of patient characteristics and mechanism of injury are responsible for the sporadic nature of ALI/ARDS and its observed phenotypic variability. Research on the pathophysiology and genetics of ALI/ARDS continues to advance, revealing critical molecular pathways in disease development and specific genetic factors that alter the expression of disease. Despite these advances, pharmacologic therapies have yet to be developed for the prevention or treatment of disease. We anticipate that continued improvement of our understanding of the genetic and pathophysiologic mechanisms underlying ALI/ARDS combined with future clinical trials will allow pharmacogenetic therapies for ALI/ARDS to be developed. PMID- 21617627 TI - Preoperative evaluation in geriatric surgery: comorbidity, functional status and pharmacological history. AB - The demand for elective and emergency surgery by older patients is increasing. This review examines the current practice of preoperative evaluation in geriatric anesthesia and provides an overview of new insights in this field. Preoperative anesthesia consultation is essential to examine the patient, evaluate the operative risk and plan preventive perioperative actions. Chronological age probably represents an independent risk factor. Age should not be considered an exclusion criterion from surgery per se. More than 50% of patients over 70 years old suffer from one infirmity, and 30% suffer from two or more infirmities. Hypertension is the most common disease, followed by coronary artery disease, diabetes and chronic obstructive pulmonary disease. Aging processes, illnesses, malnutrition, difficulties in communication and comprehension, psychological alterations and social needs may coexist and overlap. Changes in pharmacodynamics and pharmacokinetics induced by aging make elderly patients very sensitive to drugs, especially those administered perioperatively. Drug underuse, misuse and abuse are described, together with criteria to manage perioperative medications. Disability, dementia and frailty are risk factors for adverse outcomes and delirium after surgery. Traditional anesthesia consultation captures only a small portion of the necessary information, especially about functional status and frailty. Although the association between older age and surgical complications is well known, most anesthetists and surgeons do not measure physical and cognitive function preoperatively. Extending anesthesia consultation to functional status provides useful information for preoperative counseling and planning of postoperative care. A strong joint action with the surgical team is essential. Currently, while many resources are employed to assess preoperative cardiac risk and despite the dramatic increase in the number of elderly surgical patients, the association between older age itself and surgical complications has not been fully investigated, and preoperative evaluation of functional status is not yet a part of routine preoperative practice. Creating a new culture and developing appropriate clinical, scientific and relational approaches to these patients represent the core of the challenge. PMID- 21617628 TI - Ventilator-induced lung injury and sepsis: two sides of the same coin? AB - Unequivocal evidence from both experimental and clinical research has shown that mechanical ventilation can damage the lungs and initiate an inflammatory response, possibly contributing to extrapulmonary organ dysfunction. This type of injury, referred to as ventilator-induced lung injury (VILI), resembles the syndromes of acute lung injury (ALI) and the acute respiratory distress syndrome (ARDS). VILI can trigger a complex array of inflammatory mediators, resulting in a local and systemic inflammatory response. Substances produced in the lungs can be translocated into the systemic circulation as a result of injury to the pulmonary epithelium and to the capillary endothelium. This type of injury forms the basis for the use of low tidal volumes (5-7 mL/kg of predicted body weight) during mechanical ventilation of patients with ALI/ARDS. The recognition of VILI has prompted a number of investigators to suggest that ALI/ARDS may be, in part, a product of our efforts to mechanically ventilate patients rather than the progression of the underlying disease. On the other hand, current scientific evidence supports a link between VILI and the development of extrapulmonary organ dysfunction, similar to how most severe cases of sepsis are clinically manifested. In addition, functional genomics approaches using a gene array methodology to measure lung gene expression have identified differential patterns of gene expression in animal models of VILI, similar to those gene pathways activated during experimental and clinical sepsis. In this line of thought, we hypothesize that injurious mechanical ventilation could be responsible for the perpetuation and worsening of sepsis in some patients and for the development of a sepsis-like syndrome in others. PMID- 21617629 TI - Cigarette package health warnings and interest in quitting smoking --- 14 countries, 2008--2010. AB - The World Health Organization (WHO) Framework Convention on Tobacco Control (FCTC) requires health warnings on tobacco product packages sold in countries that ratified the WHO FCTC treaty (1). These warnings are expected to 1) describe the harmful effects of tobacco use; 2) be approved by the appropriate national authority; 3) appear on at least 30%, and ideally 50% or more, of the package's principal display areas; 4) be large, clear, visible, and legible in the country's principal language(s); 5) have multiple, rotating messages; and 6) preferably use pictures or pictograms. To assess the effects of cigarette package health warnings on interest in quitting smoking among smokers of manufactured cigarettes aged >=15 years, this report examines 2008--2010 data from the Global Adult Tobacco Survey (GATS) in 14 WHO FCTC countries. Among men, the prevalence of manufactured cigarette smoking ranged from 9.6% in India to 59.3% in Russia. Among men in 12 of the countries and women in seven countries, >90% of smokers reported noticing a package warning in the previous 30 days. The percentage of smokers thinking about quitting because of the warnings was >50% in six countries and >25% in men and women in all countries except Poland. WHO has identified providing tobacco health information, including graphic health warnings on tobacco packages, as a powerful "best buy" in combating noncommunicable disease (2). Implementing effective warning labels as a component of a comprehensive approach can help decrease tobacco use and its many health consequences. PMID- 21617630 TI - Human Jamestown canyon virus infection --- Montana, 2009. AB - Jamestown Canyon virus (JCV) is a mosquito-borne zoonotic pathogen belonging to the California serogroup of bunyaviruses. Although JCV is widely distributed throughout temperate North America, reports of human JCV infection in the United States are rare. This is the first report of human JCV infection detected in Montana, one of only 15 cases reported in the United States since 2004, when JCV became reportable. On May 26, 2009, a man aged 51 years with no travel history outside of Montana went to a local emergency department immediately following onset of fever, severe frontal headache, dizziness, left-sided numbness, and tingling. His blood pressure was elevated. Stroke was ruled out, oxygen was administered, medication was prescribed for hypertension, and the patient was sent home. One week later, the patient visited his primary-care physician complaining of continued neurologic symptoms consistent with acute febrile encephalitis and recent mosquito bites. Although West Nile virus (WNV) disease was diagnosed based on detection of WNV-immunoglobulin M (IgM) and G (IgG) antibodies, subsequent testing indicated that the WNV antibodies were from a past infection and that his illness was caused by JCV. The final diagnosis of JCV infection was based on positive JCV-specific IgM enzyme-linked immunosorbent assay (ELISA) results and a fourfold rise in paired sample JCV plaque reduction neutralization test (PRNT) titers. This finding represents a previously unrecognized risk for JCV infection in Montana; clinicians should consider JCV infection when assessing patients for suspected arboviral infections. PMID- 21617631 TI - Contribution of occupational physical activity toward meeting recommended physical activity guidelines: United States, 2007. AB - Regular physical activity helps maintain healthy weight and reduces the likelihood of developing chronic diseases. The 2008 Physical Activity Guidelines for Americans (1) are derived from the most recent scientific review of physical activity health benefits and do not differentiate among physical activity for leisure, transportation, work, or other purposes. To examine the potential influence of occupational physical activity on meeting minimum weekly aerobic physical activity guidelines, the Washington State Department of Health (WADOH) analyzed demographic patterns in physical activity levels with and without consideration of occupational physical activity using 2007 Behavioral Risk Factor Surveillance System (BRFSS) data. This report describes the results of that analysis, which indicated that, approximately two thirds (64.3%) of U.S. adults met minimum physical activity guidelines through nonoccupational physical activity. When occupational physical activity (defined as reported work activity of mostly walking or heavy labor) was considered, an additional 6.5% of adults likely met the guidelines. The increase was greatest for Hispanic men (14.4%) and men with less than a high school education (15.9%). Public health agencies conducting surveillance of population physical activity levels also should consider including occupational physical activity, which will help to identify demographic groups for targeted programs that increase physical activity. PMID- 21617632 TI - Recommendations for use of a booster dose of inactivated vero cell culture derived Japanese encephalitis vaccine: advisory committee on immunization practices, 2011. AB - Japanese encephalitis (JE) virus, a mosquito-borne flavivirus, is an important cause of encephalitis in Asia with a case fatality rate of 20%--30% and neurologic or psychiatric sequelae in 30%--50% of survivors (1). Travelers to JE endemic countries and laboratory personnel who work with infectious JE virus are at potential risk for JE virus infection. In 2010, CDC's Advisory Committee on Immunization Practices (ACIP) updated recommendations for prevention of JE. The updated recommendations included information on use of a new inactivated, Vero cell culture--derived JE vaccine (JE-VC [manufactured as Ixiaro]) that was licensed in the United States in 2009. Data on the need for and timing of booster doses with JE-VC were not available when the vaccine was licensed. This report summarizes new data on the persistence of neutralizing antibodies following primary vaccination with JE-VC and the safety and immunogenicity of a booster dose of JE-VC. The report also provides updated guidance to health-care personnel regarding use of a booster dose of JE-VC for U.S. travelers and laboratory personnel. ACIP recommends that if the primary series of JE-VC was administered >1 year previously, a booster dose may be given before potential JE virus exposure. PMID- 21617633 TI - Update on Japanese encephalitis vaccine for children: United States, May 2011. AB - Inactivated mouse brain--derived Japanese encephalitis (JE) vaccine (JE-MB [manufactured as JE-Vax]), the only JE vaccine that is licensed for use in children in the United States, is no longer available. This notice provides updated information regarding options for obtaining JE vaccine for U.S. children. PMID- 21617634 TI - Measles: United States, January--May 20, 2011. AB - Measles is a highly contagious, acute viral illness that can lead to serious complications and death. Endemic or sustained measles transmission has not occurred in the United States since the late 1990s, despite continued importations (1). During 2001--2008, a median of 56 (range: 37--140) measles cases were reported to CDC annually (2); during the first 19 weeks of 2011, 118 cases of measles were reported, the highest number reported for this period since 1996. Of the 118 cases, 105 (89%) were associated with importation from other countries, including 46 importations (34 among U.S. residents traveling abroad and 12 among foreign visitors). Among those 46 cases, 40 (87%) were importations from the World Health Organization (WHO) European and South-East Asia regions. Of the 118, 105 (89%) patients were unvaccinated. Forty-seven (40%) patients were hospitalized and nine had pneumonia. The increased number of measles importations into the United States this year underscores the importance of vaccination to prevent measles and its complications. PMID- 21617635 TI - Long-Term BMI changes since adolescence and markers of early and advanced subclinical atherosclerosis. AB - Although long-term weight gain has been associated with cardiovascular risk and intima-media thickening (IMT), no sufficient data exist on possible associations of such weight changes with more advanced stages of subclinical atherosclerosis. Moreover, the value of self-reported weight changes, a more practical approach to assess long-term history in adiposity status, is still a matter of debate. In this longitudinal study, long-term changes in BMI and overweight status were assessed in 106 healthy young adults (age 40.5 +/- 1.1 years, 60 males). These were a subgroup of adolescent school students who had originally been examined in 1983 initially aiming to assess cardiovascular risk factor prevalence. Markers of early (carotid IMT) and advanced (presence of plaques in the carotid and femoral arteries and ankle-brachial index, ABI) subclinical atherosclerosis were measured in all individuals. By multivariate analysis, among other risk factors, IMT and the presence of plaques were independently determined by BMI change, while a low ABI was also determined by changes in overweight status. An adverse long term adiposity profile change (>= +4 kg/m(2) and/or change into overweight/obese status from normal weight since adolescence) incrementally determined a low ABI over current risk factors. Self-reported and actual BMI changes were correlated (r = 0.587) but their means significantly differed, while the former significantly correlated with IMT only (P = 0.032). In conclusion, an adverse long term adiposity status change was more prominently associated with advanced subclinical atherosclerosis and particularly low ABI. These results also suggest that the utility of self-reported weight changes may be limited in primary prevention practice. PMID- 21617636 TI - Time use and food pattern influences on obesity. AB - The rise of obesity in the United States over the past 25 years has resulted in an increase in the number of research studies published related to the causes, consequences, and possible solutions to the problem. Most would agree that obesity is a multi-dimensional problem that requires a range of solutions related to individual diet and activity, food and built environment, and public policy. Examination of complex relationships between food choice, time use patterns, sociodemographic characteristics and obesity has been limited by data availability and disciplinary focus. Using the theory of the production of health capital, this paper links empirical data from the Consumer Expenditure, Current Population, and American Time Use Surveys to provide estimates of the impacts of food expenditure and time use patterns on obesity in single female headed households of 31-50 years of age. PMID- 21617637 TI - Tobacco smoke exposure before, during, and after pregnancy and risk of overweight at age 6. AB - Maternal smoking during pregnancy has been associated with overweight and obesity in childhood and is strongly correlated with children's tobacco smoke exposure before and after pregnancy. We investigated the independent association of tobacco smoke exposure at various pre- and postnatal periods and overweight at age 6. A total of 1,954 children attending the 2001-2002 school entrance health examination in the city of Aachen, Germany, were included into this study. Height and weight were measured, BMI was calculated. Tobacco smoke exposure at various periods, other lifestyle and sociodemographic factors were ascertained by questionnaire. Multiple logistic regression models were used to assess the association between tobacco smoke exposure and overweight. Prevalence of overweight was 8.9%. Significant positive associations were found with maternal smoking before and during pregnancy and during the first and sixth year of life. When all smoking periods were included into one logistic model simultaneously, secondhand smoke exposure after birth remained positively associated with overweight at age 6 at either one of the two time periods (first year only: odds ratio (OR) (95% confidence interval (CI)): 2.94 (1.30-6.67), sixth year only: 2.57 (1.64-4.04), respectively) or at both (4.43 (2.24-8.76)). Exposure to tobacco smoke during the first years of life appears to be a key risk factor for development of childhood overweight. PMID- 21617638 TI - Effects of smoking and obesity on the association between CDH13 (rs3865188) and adiponectin among Korean men: the KARE study. AB - Adiponectin is secreted exclusively by adipocytes. Adiponectin is regulated by obesity, smoking, and genetic factors, including CDH13, which may contribute to development of diseases such as cardiovascular disease. Therefore, we aim to explore the joint effect of smoking and obesity on the association between CDH13 (rs3865188) and adiponectin among Korean men. This study included 1,570 Korean men aged 40-69 years who participated in the KARE cohort study (community-based cohorts, South Korea) from 2001. Hypoadiponectinemia was defined as the lowest quartile of adiponectin. In this study, individuals with at least one at-risk allele, the T allele, had an increased risk for hypoadiponectinemia, particularly current smokers with at least one T allele together with obesity when compared to those without the T allele. In addition, individuals with the TT genotype of CDH13 rs3865188, as well as obesity, were significantly associated with a 1.8 fold (odds ratio (OR) = 9.4, 95% confidence interval (CI) = 4.4-19.6, P < 0.001) increased risk for hypoadiponectinemia compared to individuals with the TT genotype of CDH 13 rs3865188 and normal waist circumference (WC) (OR = 5.1, 95% CI = 3.5-7.6, P < 0.001). However, in the joint effect of smoking and obesity, current smokers with the TT genotype of CDH13 rs3865188, as well as obesity, were significantly associated with a 6.2-fold (OR = 24.2, 95% CI = 3.0-196.6, P < 0.001) increased risk for hypoadiponectinemia compared to nonsmokers with the TT genotype of CDH 13 rs3865188 with normal WC (OR = 3.9, 95% CI = 1.7-9.3, P < 0.001). This study suggested that the association between CDH13 and adiponectin can be modified by lifestyle factors, such as smoking and obesity, among Korean men. PMID- 21617640 TI - The relationship of birth weight with longitudinal changes in body composition in adult women. AB - Most research on birth weight and adult health status has reported adult measures at a single time point. This study examined the relationship of self-reported birth weight to longitudinal changes in adult body composition in 587 women of the Michigan Bone Health and Metabolism Study, followed from 1992 to 2007 and aged 24-50 years at baseline. Linear mixed models were used to estimate the association between three birth weight categories and women's 15-year changes in adult weight, height, BMI, waist and hip circumference, waist-to-hip ratio, and fat, lean, and skeletal muscle mass. Body composition measures increased in all women over the 15-year study period. At their adult baseline, high birth weight women weighed 13% more and had waist circumference and lean mass measures that were 5.51 cm and 3.91 kg larger, respectively, than normal birth weight women. No differences were observed in adult body composition between low and normal birth weight women and rates of change in the adult measures did not vary across the birth weight groups. Women heavier at birth continued to be heavier through adulthood, corroborating previous reports based on single measures of adult body composition. Research to address whether higher adult body composition in high birth weight women increases the longitudinal risk for obesity-related chronic diseases is needed. PMID- 21617639 TI - Lower extremity fat mass is associated with insulin resistance in overweight and obese individuals: the CARDIA study. AB - Lower extremity fat mass (LEFM) has been shown to be favorably associated with glucose metabolism. However, it is not clear whether this relationship is similar across varying levels of obesity. We hypothesized that lower amounts of LEFM is associated with higher insulin resistance (IR) and this association may vary according to weight status. Participants with available measures were examined from the Coronary Artery Risk Development in Young Adults study (CARDIA), a multi center longitudinal study of the etiology of atherosclerosis in black and white men and women aged 38-50 years old in 2005-2006 (n = 1,579). The homeostasis model assessment of IR (HOMA(IR)) was calculated to estimate IR, regional adiposity was measured using dual energy X-ray absorptiometry (DXA), and weight status was defined according to BMI categories. Obese and overweight participants exhibited higher IR, total fat mass (FM), trunk FM (TFM), and LEFM compared to normal weight participants. After controlling for age, height, race, study center, education, smoking, and cardiorespiratory fitness (CRF), greater LEFM was significantly associated with higher IR only in normal weight men and women. Further adjustment for TFM revealed that lower LEFM was significantly associated with higher IR in overweight and obese men and women and the positive association in normal weight individuals was attenuated. These results suggest that excess adiposity in the lower extremities may attenuate the metabolic risk observed at a given level of abdominal adiposity in overweight and obese individuals. Weight status presents additional complexity since the metabolic influence of adipose tissue may not be homogenous across anatomic regions or level of obesity. PMID- 21617641 TI - Increased PTH and 1.25(OH)(2)D levels associated with increased markers of bone turnover following bariatric surgery. AB - The objective of this study was to characterize changes in metabolic bone parameters following bariatric surgery. Seventy-three obese adult patients who underwent either gastric banding (GB), Roux-en-Y gastric bypass (RYGB), or biliopancreatic diversion with duodenal switch (BPD/DS) were followed prospectively for 18 months postoperatively. Changes in the calcium-vitamin D axis (25-hydroxyvitamin D (25OHD), 1,25-dihydroxyvitamin D (1,25(OH)(2)D), calcium, parathyroid hormone (PTH)), markers of bone formation (osteocalcin, bone specific alkaline phosphatase) and resorption (urinary N-telopeptide (NTx)), as well as bone mineral density (BMD) were assessed at 3-month intervals during this time period. Bariatric surgery resulted in significant and progressive weight loss over 18 months. With supplementation, 25OHD levels increased 65.3% (P < 0.0001) by 3 months, but leveled off and decreased <30 ng/ml by 18 months. PTH initially decreased 21.4% (P = 0.01) at 3 months, but later approached presurgery levels. 1,25(OH)(2)D increased significantly starting at month 12 (50.3% increase from baseline, P = 0.008), and was positively associated with PTH (r = 0.82, P = 0.0001). When stratified by surgery type, median PTH and 1,25(OH)(2)D levels were higher following combined restrictive and malabsorptive operations (RYGB and BPD/DS) compared to GB. Bone formation/resorption markers were increased by 3 months (P < 0.05) and remained elevated through 18 months. Radial BMD decreased 3.5% by month 18, but this change was not significant (P = 0.23). Our findings show that after transient improvement, preoperative vitamin D insufficiency and secondary hyperparathyroidism persisted following surgery despite supplementation. Postoperative secondary hyperparathyroidism was associated with increased 1,25(OH)(2)D levels and increased bone turnover markers. PMID- 21617642 TI - Nutritional strategies and growth in extremely low birth weight infants with bronchopulmonary dysplasia over the past 10 years. AB - OBJECTIVE: Changes in nutritional strategies over the past decade have been shown to improve postnatal growth in extremely low birth weight (ELBW) infants. We showed 10 years ago that the majority of these ELBW infants with bronchopulmonary dysplasia (BPD) suffer postnatal growth failure. We theorized that recent changes in nutritional support strategies would positively affect growth outcomes in ELBW infants with BPD. STUDY DESIGN: A retrospective study of 88 ELBW infants with BPD. Nutritional data, postnatal growth and BPD severity were compared across three cohorts: (1) weight gain <=14 g kg(-1) per day, (2) 14.1 to 16 g kg(-1) per day and (3) >=16 g kg(-1) per day from return to birth weight through discharge. We also compared these to a historical cohort. RESULT: In all, 73% of current subjects grew at or above fetal rates. There was less extrauterine growth restriction (EUGR) by weight and head circumference for those ELBW infants with BPD receiving higher amounts of protein. Aggressive early TPN and receipt of caloric-dense milk seemed to be the 'new' nutritional strategies improving growth for current ELBW infants with BPD compared with those 10 years ago. CONCLUSION: Despite a diagnosis of BPD, improved nutritional strategies have enhanced postnatal growth in infants at high risk for EUGR. PMID- 21617643 TI - The use of mobile robotic telemedicine technology in the neonatal intensive care unit. AB - OBJECTIVE: To describe the use of a wireless, mobile, robotic telecommunications system in the Neonatal Intensive Care Unit (NICU). STUDY DESIGN: In this prospective study utilizing 304 patient encounters on 46 preterm and term neonates in a level IIIa NICU, a bedside neonatologist ('on-site neonatologist'; ONSN) and a neonatologist at a distant location ('off-site neonatologist'; OFFSN) evaluated selected demographic information, laboratory data and clinical and radiological findings of the subjects. The OFFSN used a commercial wireless, mobile, robotic telecommunications system controlled from a remote site. The two physicians were blinded to each other's findings and agreement rates of the evaluations between the ONSN and the OFFSN were compared using kappa statistics. Agreement rates between two ONSNs using the same protocol with 39 patient encounters served as the reference standard. The dependability and timeliness of data transmission were also assessed. RESULT: Excellent or intermediate-to-good agreements were noted for all but a few physical examination assessments between both the ONSN and OFFSN and the two ONSNs. Poor agreements were found for certain physical examination parameters (breath-, heart- and bowel-sounds and capillary refill time) with or without the use of telemedicine. The median duration of the encounters by the ONSN and OFFSN and the two ONSNs was similar. Five encounters were excluded from the analysis because of technical difficulties. No complications associated with the use of the mobile robot were noted. CONCLUSION: Our findings indicate that the use of mobile robotic telemedicine technology is feasible for neonates in the NICU. PMID- 21617646 TI - Uropathogenic bacteria leave a mark. AB - Urinary tract infections are common, obnoxious and in some cases even life endangering. They are most often caused by uropathogenic Escherichia coli. Persistent and seemingly recurrent infections may be caused by bacteria establishing intracellular reservoirs in the urothelial epithelium. A better understanding of the mechanisms involved in the complex interplay between host and pathogens is required for preventing and controlling these infections. A paper in this issue of Laboratory Investigation reports changes in gene expression in urothelial carcinoma cells following infection by uropathogenic bacteria which go along with the activation of DNA methyltransferase 1 and increased methylation at the CDKN2A gene. These pioneer findings should stimulate the further development of in vitro models for studying urothelial infections and prompt more extensive analyses of epigenetic alterations elicited by bacterial infections in the urinary tract. PMID- 21617644 TI - Induction of human neuronal cells by defined transcription factors. AB - Somatic cell nuclear transfer, cell fusion, or expression of lineage-specific factors have been shown to induce cell-fate changes in diverse somatic cell types. We recently observed that forced expression of a combination of three transcription factors, Brn2 (also known as Pou3f2), Ascl1 and Myt1l, can efficiently convert mouse fibroblasts into functional induced neuronal (iN) cells. Here we show that the same three factors can generate functional neurons from human pluripotent stem cells as early as 6 days after transgene activation. When combined with the basic helix-loop-helix transcription factor NeuroD1, these factors could also convert fetal and postnatal human fibroblasts into iN cells showing typical neuronal morphologies and expressing multiple neuronal markers, even after downregulation of the exogenous transcription factors. Importantly, the vast majority of human iN cells were able to generate action potentials and many matured to receive synaptic contacts when co-cultured with primary mouse cortical neurons. Our data demonstrate that non-neural human somatic cells, as well as pluripotent stem cells, can be converted directly into neurons by lineage determining transcription factors. These methods may facilitate robust generation of patient-specific human neurons for in vitro disease modelling or future applications in regenerative medicine. PMID- 21617648 TI - Dental hygienists and therapists: how much professional autonomy do they have? How much do they want? Results from a UK survey. AB - OBJECTIVES: To investigate autonomous working among singly and dually qualified dental hygienists and therapists in UK primary care. Earlier studies and policy papers suggest that greater autonomy for these groups may be a desirable workforce planning goal. METHODS: UK-wide postal surveys of hygienists, hygienist therapists and therapists. Respondents were asked whether they undertook 15 clinical activities on their own initiative, how comfortable they would feel undertaking such clinical activities if referral from a dentist were not required, and how they perceived dentists' reactions. RESULTS: Overall response rate was 65% (n = 150 hygienists, 183 hygienist-therapists and 152 therapists). Over 80% of hygienists and hygienist-therapists reported undertaking BPEs, history-taking, pocket charting, mucosal examinations and recall interval planning autonomously. Similarly high proportions of hygienist-therapists and therapists reported giving local analgesia and choosing restorative materials autonomously. However, fewer than 50% of all three groups said they undertook dental charting, fissure sealing, resin restorations, taking radiographs, and tooth whitening autonomously. While confidence in undertaking such activities without a dentist's referral was generally high, it was lower in respect to mucosal examinations, identifying suspicious lesions, interpreting radiographs, tooth whitening, and (except for singly qualified dental therapists) diagnosing caries. CONCLUSIONS: Results suggest high levels of experience and confidence in their ability to work autonomously across a wide range of investigative activities, treatment decision-making and treatment planning. The exceptions to this pattern are appropriate to the different clinical remit of these groups. PMID- 21617650 TI - Presidential address. PMID- 21617649 TI - Modelling the impact of process variables in community fluoridated milk schemes on a population of UK schoolchildren. AB - BACKGROUND: Dental caries is a public health problem. Fluoridated milk (FM) schemes are used as a preventive measure. The impact of process variables in these schemes is not understood. METHODS: Process variable data on the number of days of consumption, attendance, volume consumed, parental consent together with the proportion of children drinking FM at 7- and 11-years old were aggregated from eight schemes in the UK. The impact of process variables was modelled in an 'averaged' scheme (reduced in effectiveness by process variables) and compared with a notional 'ideal' one in which no process variables operate. Parental consent was analysed according to socio-economic groupings. RESULTS: Proportion of days per year FM was consumed: 0.52. Values for process variables were: attendance rate 0.94; proportion of milk consumed 0.91; proportion of children with parental consent at 5 years 0.65; proportion drinking FM at 7 and 11 years respectively 0.54 and 0.27. No clear trends were observed for parental consent across socio-economic groupings. CONCLUSION: Modelling suggests that due to the cumulative impact of process variables, there is cause for concern about the effectiveness of FM schemes as currently managed in the UK as a standalone public health measure for the prevention of caries. PMID- 21617651 TI - Valedictory address. PMID- 21617652 TI - OMFS team snookered. PMID- 21617653 TI - Plea for transparency. PMID- 21617654 TI - Sound via teeth. PMID- 21617655 TI - Animal ethics. PMID- 21617656 TI - Cleft pathways. PMID- 21617670 TI - Summary of: Dental hygienists and therapists: how much professional autonomy do they have? How much do they want? Results from a UK survey. PMID- 21617671 TI - Summary of: Modelling the impact of process variables in community fluoridated milk schemes on a population of UK schoolchildren. PMID- 21617672 TI - Preparing dental students for careers as independent dental professionals: clinical audit and community-based clinical teaching. AB - Community-based clinical teaching programmes are now an established feature of most UK dental school training programmes. Appropriately implemented, they enhance the educational achievements and competences achieved by dental students within the earlier part of their developing careers, while helping students to traverse the often-difficult transition between dental school and vocational/foundation training and independent practice. Dental school programmes have often been criticised for 'lagging behind' developments in general dental practice - an important example being the so-called 'business of dentistry', including clinical audit. As readers will be aware, clinical audit is an essential component of UK dental practice, with the aims of improving the quality of clinical care and optimising patient safety. The aim of this paper is to highlight how training in clinical audit has been successfully embedded in the community-based clinical teaching programme at Cardiff. PMID- 21617673 TI - Evaluation of a community-based clinical teaching programme by current and former student dental therapists and dental hygienists: a pilot investigation. AB - AIM: There has been considerable expansion in the involvement of community-based clinical teaching programmes (sometimes termed 'outreach teaching') in UK and other international dental schools. While there has been much interest in the role of this educational methodology in the professional and educational development of student dentists, there has been little, if no, consideration of this form of teaching in relation to dental care professional (DCP) students. The aim of this pilot investigation was to report the feedback and evaluation of current and former student dental therapists and dental hygienists on their experience on the St David's community-based clinical teaching programme at Cardiff. METHODS: In Autumn 2009, a questionnaire was distributed by hand to the current second year student dental therapist and dental hygiene class at Cardiff (n = 18) and by post to the dental therapist and dental hygiene classes of 2004 (n = 16) and 2007 (n = 17). The questionnaire included both 'open' and 'closed' questions. RESULTS: Thirty responses were returned (response rate = 59%; 2004 (n = 5, 31%), 2007 (n = 9, 53%), current class (n = 16, 89%)). Seventy percent of respondents (n = 21) reported that they found the community-based clinical teaching programme to be a pleasant working environment and close to subsequent independent practice. Seventy-seven percent (n = 23) reported that their confidence performing nonsurgical periodontal treatment had increased while at the programme. One respondent commented that the programme was '...an invaluable and insightful introduction to what it would be like working in practice. Without being given the experience, it would have been a big shock to the system when I started working in practice...' CONCLUSION: This pilot investigation has revealed that current and former dental therapist and dental hygiene students are enthusiastic in their support for the inclusion of community-based clinical teaching programmes in their educational and professional development. Most former and current dental therapist and dental hygiene students noted the positive effects of this form of training on their subsequent clinical careers. PMID- 21617691 TI - Antithrombotic therapy. Lancelot results published. PMID- 21617692 TI - NLRX1/NOD5 deficiency does not affect MAVS signalling. PMID- 21617693 TI - Epithelial cell polarity: a major gatekeeper against cancer? AB - The correct establishment and maintenance of cell polarity are crucial for normal cell physiology and tissue homeostasis. Conversely, loss of cell polarity, tissue disorganisation and excessive cell growth are hallmarks of cancer. In this review, we focus on identifying the stages of tumoural development that are affected by the loss or deregulation of epithelial cell polarity. Asymmetric division has recently emerged as a major regulatory mechanism that controls stem cell numbers and differentiation. Links between cell polarity and asymmetric cell division in the context of cancer will be examined. Apical-basal polarity and cell-cell adhesion are tightly interconnected. Hence, how loss of cell polarity in epithelial cells may promote epithelial mesenchymal transition and metastasis will also be discussed. Altogether, we present the argument that loss of epithelial cell polarity may have an important role in both the initiation of tumourigenesis and in later stages of tumour development, favouring the progression of tumours from benign to malignancy. PMID- 21617694 TI - The golden anniversary of the thymus. AB - The immunological function of the thymus was first documented 50 years ago by using neonatally thymectomized mice, while studying its role in virus-induced leukaemia. Since then, an enormous wealth of reports has helped to define the importance of this primary lymphoid organ. In this article, I summarize the key advances that have led to our current knowledge of the functions of the thymus and its T cells in immunity. PMID- 21617695 TI - Short-term changes in the optic nerve head and visual field after trabeculectomy. AB - AIMS OR PURPOSE: To evaluate short-term changes in optic nerve head topography and visual field induced by surgical reduction of intraocular pressure. METHODS: A prospective study was performed on 56 eyes of 56 patients with uncontrolled primary open-angle glaucoma despite maximum medical therapy, which underwent trabeculectomy. Optic nerve head evaluations by means of Heidelberg Retina Tomograph, and visual field tests were performed pre-operatively, and at 3 and 6 months after surgery. Differences in intraocular pressure, visual field indices, and Heidelberg Retina Tomograph parameters were evaluated with the t-test for paired data. A linear regression model was calculated to analyze the relationship between intraocular pressure reduction and visual field changes, and optic nerve head changes. RESULTS: Mean intraocular pressure decreased from 24.4 +/- 5.0 mm Hg to 12.1 +/- 3.1 mm Hg (month 3, P<0.001), and 10.6 +/- 2.8 mm Hg (month 6, P<0.001) after trabeculectomy. Mean retinal nerve fiber layer thickness (baseline, 0.19 +/- 0.034; month 3, 0.24 +/- 0.039, P=0.05; month 6, 0.21 +/- 0.037, P=0.05) showed a statistical significant change compared with baseline values. CONCLUSIONS: In this 6-months study, a significant increase in retinal nerve fiber layer thickness was detected after glaucoma filtration surgery. PMID- 21617696 TI - Anatomical and functional outcomes in contusion injuries of posterior segment. AB - PURPOSE: To evaluate the clinical features, and anatomical and visual outcomes in patients with closed-globe contusion injury involving the posterior segment. METHODS: Retrospective review of posterior segment contusion injuries admitted to our tertiary referral center. RESULTS: In all, 115 patients (115 eyes) with complete data were reviewed. Surgery had been performed in 79 (69%) patients. The mean follow-up period was 6 months (range, 2-34 months). Retinal detachment, in 31% of eyes, was the most frequently encountered posterior segment pathology. The presence of retinal detachment was associated with poor visual outcome (<20/100), (P<0.001). Coexisting (five patients, 4%) and postoperative proliferative vitreoretinopathy (PVR) (two patients, 2%) was the main cause of failure in these cases. A significant positive correlation was obtained between initial and final visual acuity levels in both the medical treatment group and the surgical treatment group (P<0.05). The presenting visual acuity of <20/400 was associated with poor visual outcome (P<0.05 for both groups). Poor visual outcome in 13 patients with successful repair of retinal detachment was due to the macular lesions and the optic atrophy. CONCLUSION: Retinal detachment was the most frequently encountered posterior segment pathology subsequent to closed-globe contusion injuries. In addition to macular scarring and optic nerve damage, development of PVR has prognostic significance in these eyes. PMID- 21617697 TI - Mesenchymal stromal cell 'licensing': a multistep process. AB - Many in vitro and in vivo data are available supporting the role of mesenchymal stromal cell (MSC) licensing in the induction of a measurable and effective immune regulation. The failure of some MSC-based protocols for immune modulation in animal models and in human clinical trials may be explained by either lack of a proper licensing by inflammatory microenvironment or wrong timing in MSC administration. Thus, optimization of MSC use for immune-regulating purposes is required to maximize their beneficial effects. PMID- 21617699 TI - TLR4 knockout protects mice from radiation-induced thymic lymphoma by downregulation of IL6 and miR-21. PMID- 21617698 TI - Immunophenotypic and gene expression analysis of monoclonal B-cell lymphocytosis shows biologic characteristics associated with good prognosis CLL. AB - Monoclonal B-cell lymphocytosis (MBL) is a hematologic condition wherein small B cell clones can be detected in the blood of asymptomatic individuals. Most MBL have an immunophenotype similar to chronic lymphocytic leukemia (CLL), and 'CLL like' MBL is a precursor to CLL. We used flow cytometry to identify MBL from unaffected members of CLL kindreds. We identified 101 MBL cases from 622 study subjects; of these, 82 individuals with MBL were further characterized. In all, 91 unique MBL clones were detected: 73 CLL-like MBL (CD5(+)CD20(dim)sIg(dim)), 11 atypical MBL (CD5(+)CD20(+)sIg(+)) and 7 CD5(neg) MBL (CD5(neg)CD20(+)sIg(neg)). Extended immunophenotypic characterization of these MBL subtypes was performed, and significant differences in cell surface expression of CD23, CD49d, CD79b and FMC-7 were observed among the groups. Markers of risk in CLL such as CD38, ZAP70 and CD49d were infrequently expressed in CLL-like MBL, but were expressed in the majority of atypical MBL. Interphase cytogenetics was performed in 35 MBL cases, and del 13q14 was most common (22/30 CLL-like MBL cases). Gene expression analysis using oligonucleotide arrays was performed on seven CLL-like MBL, and showed activation of B-cell receptor associated pathways. Our findings underscore the diversity of MBL subtypes and further clarify the relationship between MBL and other lymphoproliferative disorders. PMID- 21617700 TI - Refining the diagnosis of T-cell large granular lymphocytic leukemia by combining distinct patterns of antigen expression with T-cell clonality studies. AB - T-cell large granular lymphocytic (LGL) leukemia is a complex diagnosis, requiring persistent clonal expansions of LGLs, and cytopenias. Often the diagnosis is unclear as non-clonal expansions of LGLs commonly occur in reactive conditions. To better understand T-LGL leukemia, we performed a comprehensive clinicopathologic analysis of 85 patients with LGL expansions. Interestingly, distinct CD8+(dim)/CD57+ populations, seen by flow cytometry, were significantly associated with clonal T-LGL leukemia (P < 0.001) as well as neutropenia (median absolute neutrophil count (ANC) 1.45 vs 3.19 * 10(9)/l; P = 0.0017). Furthermore, cases with distinct CD8+(dim)/CD57+ populations and monoclonal T cells had even lower ANCs (median ANC 1.41 * 10(9)/l; P = 0.001) compared with cases without these dual criteria. Additionally, complete or partial loss of CD5 expression was independently associated with clonal T-LGL leukemia (P<0.001) and neutropenia (median ANC 1.41 vs 2.70 * 10(9)/l; P = 0.002). This study describes specific immunophenotypic parameters to better define clonal cases of T-LGL leukemia associated with significant neutropenia. PMID- 21617701 TI - Correction of murine Rag1 deficiency by self-inactivating lentiviral vector mediated gene transfer. AB - Severe combined immunodeficiency (SCID) patients with an inactivating mutation in recombination activation gene 1 (RAG1) lack B and T cells due to the inability to rearrange immunoglobulin (Ig) and T-cell receptor (TCR) genes. Gene therapy is a valid treatment option for RAG-SCID patients, especially for patients lacking a suitable bone marrow donor, but developing such therapy has proven challenging. As a preclinical model for RAG-SCID, we used Rag1-/- mice and lentiviral self inactivating (SIN) vectors harboring different internal elements to deliver native or codon-optimized human RAG1 sequences. Treatment resulted in the appearance of B and T cells in peripheral blood and developing B and T cells were detected in central lymphoid organs. Serum Ig levels and Ig and TCR Vbeta gene segment usage was comparable to wild-type (WT) controls, indicating that RAG mediated rearrangement took place. Remarkably, relatively low frequencies of B cells produced WT levels of serum immunoglobulins. Upon stimulation of the TCR, corrected spleen cells proliferated and produced cytokines. In vivo challenge resulted in production of antigen-specific antibodies. No leukemia development as consequence of insertional mutagenesis was observed. The functional reconstitution of the B- as well as the T-cell compartment provides proof-of principle for therapeutic RAG1 gene transfer in Rag1-/- mice using lentiviral SIN vectors. PMID- 21617702 TI - Novel H1N1 influenza in neonates: from mild to fatal disease. AB - Analysis of pediatric deaths associated with pandemic A H1N1 influenza shows that fatal outcome is more likely in young children, under the age of 5. Neonates, because of the immaturity of their immune system, could represent a high-risk group for severe disease and fatal outcome. We present a group of five neonates with confirmed novel influenza A H1N1 infection. This report indicates that the full spectrum of influenza A H1N1 infection ranging from mild febrile illness with spontaneous recovery to severe disease with fatal outcome may be expected even in neonates. PMID- 21617703 TI - A member of the ETS family, EHF, and the ATPase RUVBL1 inhibit p53-mediated apoptosis. AB - Tumour cells are known to be dependent on, or 'addicted to', not only oncogenes, but also some non-oncogenes. However, the mechanisms by which tumour cells are addicted to these genes have not been fully explained. Here, we show that overexpression of a member of the ETS family, EHF, is required for the survival of colon tumour cells that contain wild-type p53. We found that EHF directly activates the transcription of RUVBL1, an ATPase associated with chromatin remodelling complexes. RUVBL1 blocks p53-mediated apoptosis by repressing the expression of p53 and its target genes. Moreover, we found that RUVBL1 represses p53 transcription by binding to the p53 promoter, interfering with RNF20/hBRE1 mediated histone H2B monoubiquitination and promoting PAF1-mediated histone H3K9 trimethylation. These results indicate that EHF-mediated RUVBL1 expression allows colon tumour cells to avoid p53-mediated apoptosis. Thus, EHF and RUVBL1 might be promising molecular targets for the treatment of colon tumours. PMID- 21617704 TI - E-cadherin is crucial for embryonic stem cell pluripotency and can replace OCT4 during somatic cell reprogramming. AB - We report new functions of the cell-adhesion molecule E-cadherin in murine pluripotent cells. E-cadherin is highly expressed in mouse embryonic stem cells, and interference with E-cadherin causes differentiation. During cellular reprogramming of mouse fibroblasts by OCT4, SOX2, KLF4 and c-MYC, fully reprogrammed cells were exclusively observed in the E-cadherin-positive cell population and could not be obtained in the absence of E-cadherin. Moreover, reprogrammed cells could be established by viral E-cadherin in the absence of exogenous OCT4. Thus, reprogramming requires spatial cues that cross-talk with essential transcription factors. The cell-adhesion molecule E-cadherin has important functions in pluripotency and reprogramming. PMID- 21617705 TI - Targeted therapy in head and neck cancer. AB - AIMS AND BACKGROUND: This review focuses on recent advances in understanding the molecular mechanisms at the basis of cancer initiation and progression in the head and neck and also discusses the possible development of targeted cellular strategies. Intrinsic and acquired resistance of cancer cells to current conventional treatments, as well as recurrence, represent a major challenge in treating and curing the most aggressive and metastatic tumors also in the head and neck. Even though in some hematologic malignancies (i.e., non-Hodgkin's lymphomas) antibodies specifically designed to target tumor-specific cells have already been introduced, in solid tumors molecular targeted therapy is now entering clinical practice. METHODS: A PubMed database systematic review. RESULTS AND CONCLUSIONS: Molecular targeting could achieve specific damage to cancer cells, at the same time preserving functionally important tissues. This could offer new prospectives in primary and adjuvant treatment also of head and neck tumors. PMID- 21617706 TI - Do pre-diagnostic drinking habits influence breast cancer survival? AB - AIMS AND BACKGROUND: Alcohol consumption increases the risk of developing breast cancer and may also be associated with late diagnosis, recurrence, distant metastases and death. Many studies have examined the role of alcohol as a risk factor for the development of breast cancer, but very few studies have addressed the role of alcohol as a prognostic factor for survival among women diagnosed with breast cancer. The aim of this study was to investigate the survival of women with breast cancer in relation to pre-diagnostic alcohol intake and other factors known to influence prognosis. METHODS: We analyzed data for 264 women in the EUROCARE and ORDET studies who were diagnosed with breast cancer from 1987 up to 31 December 2001 and for whom information was available on follow-up, stage at diagnosis, HER-2 and hormone receptor status, and pre-diagnostic dietary alcohol intake, categorized as zero (0 g/day, non-drinkers), moderate (up to 13 g/day, about 1 serving) and high (>13 g/day). Ten-year relative survival was estimated using the maximum-likelihood approach. The excess risk of death within 10 years of diagnosis was modeled by level of alcohol intake, adjusting separately for age, stage, body mass index and tumor subtype. RESULTS: Ten-year relative survival was lower in women who drank more than 13 g/day (65%; 95% CI, 47-78) than in non-drinkers (88%; 95% CI, 75-95). The excess risk of death within 10 years was significantly higher in women who drank more than 13 g/day than non drinkers (relative excess risk, 4.13; 95% CI, 1.69-10.10) and was not altered by adjustment for other prognostic factors. The excess risk within 10 years was higher for women with a body mass index of 25 kg/m2 or higher (relative excess risk, 2.20; 95% CI, 1.01-4.70) and higher for those with more advanced disease. CONCLUSIONS: Women who drank more than 13 g alcohol per day had lower survival than non-drinkers. The excess risk of death within 10 years of diagnosis was unaffected by other known risk factors. High alcohol consumption may be an adverse prognostic factor for breast cancer. PMID- 21617707 TI - Trends in prostate cancer incidence and mortality before and after the introduction of PSA testing in the Slovak and Czech Republics. AB - AIMS AND BACKGROUND: As two neighboring countries in central Europe with national cancer registries, the Slovak (SR) and Czech Republics (CR) are countries with a medium global rate in the occurrence of prostate cancer. This paper analyzes the incidence of prostate cancer and mortality before and after the introduction of PSA testing in the two Republics and the possible reasons for any differences discovered and compares the results with selected regions and countries of the world. STUDY DESIGN AND RESULTS: In the Slovak Republic, prostate cancer incidence (age-adjusted to the world standard population) has risen from 14.6/100,000 in 1968 (95% CI, +/-1.5772) to 36.2/100,000 in 2005 (95% CI, +/ 2.0678). The estimated annual increase in the incidence during the period 1968 1991 (before nationwide PSA testing) was 0.421; from 1991 (when nationwide PSA testing began) to up to 2003 it was 0.941. Mortality rates grew from 7.3/100,000 in 1968 to 14.9/100,000 in 2005. In spite of the geographic proximity of the two countries, the increase in incidence occurred faster in the Czech than in the Slovak Republic, from 15.8/100,000 in 1977 (95% CI, +/-0.9748) to 59.5/100,000 in 2005 (95% CI, +/-1.7187). The estimated annual increase in incidence in the Czech Republic for the period of 1977-1991 was 0.581. From 1991 (when national PSA testing began) until 2003, it was 1.981. In the period before 1991, mortality rose more sharply in the Czech than in the Slovak Republic, whereas after the introduction of PSA testing mortality stabilized more quickly in the Czech than in the Slovak Republic. In the Slovak Republic, a significant reduction in mortality was observed after 2002 and has continued to the present and probably is not affected only by the results connected with the increase in PSA testing. CONCLUSIONS: The difference in the incidence and mortality of prostate cancer in the Slovak and the Czech Republics results from a difference in the intensity of PSA testing as well as from the introduction of complex, more effective treatment in advanced clinical stages. PMID- 21617708 TI - Malignant mesothelioma of the pleura in nonagenarian patients. AB - AIMS AND BACKGROUND: Malignant mesothelioma developing at very old ages is a rare event. The reasons for such late development were investigated. METHODS: A series of 811 malignant mesothelioma of the pleura, diagnosed at the Trieste and Monfalcone Hospitals, in northeastern Italy, in the period 1968-2008 were reviewed. Eight cases regarding patients aged 90 years or more were selected. In such cases, occupational histories were re-examined, and additional data could be obtained from the patients' relatives. Routine lung sections obtained at necropsy were examined for asbestos bodies. In 2 cases, asbestos bodies had been isolated after chemical digestion of lung tissue. RESULTS: The group included 7 men and one woman, aged between 90 and 93 years. All 8 patients had long-term histories of occupational exposure to asbestos, mostly in shipyards. Latency periods, elapsed between first exposure to asbestos and tumor manifestation, ranged between 64 and 75 years. Asbestos bodies were found on routine lung sections in 6 cases. Isolation of lung asbestos bodies showed 72,000 bodies per gram of dried tissue in a 90-year-old man, who had worked in the shipyards for 34 years, and 150 bodies per gram in a 93-year-old woman, who had worked in the shipyards for 23 years. CONCLUSIONS: In this group of cases, the late development of mesothelioma can not be attributed to mild exposure to asbestos or to unusually late exposures. Very long latency periods even in people heavily exposed suggest an individual resistance to the oncogenic effects of asbestos. PMID- 21617710 TI - Infantile fibrosarcoma: retrospective analysis of eleven patients. AB - AIMS AND BACKGROUND: Infantile fibrosarcomas are soft tissue sarcomas that are diagnosed at or soon after birth. In the present study, we retrospectively evaluated clinical characteristics, treatment modalities and outcome of patients diagnosed with infantile fibrosarcoma at our institution. METHODS: A retrospective review was conducted to evaluate demographic characteristics, presenting features, type and timing of surgery, other treatment modalities and survival characteristics. RESULTS: Nine males and 2 females were diagnosed with infantile fibrosarcoma between 1970-2008. The initial surgical procedure was subtotal resection in 4 patients, gross-total resection in 3 and biopsy in 4. Neoadjuvant chemotherapy was given to 10 patients. Three patients died, one for the disease and 2 from complications of therapy. Eight patients are under follow up with no evidence of disease for 1.3 to 13.5 years. None of the patients in the series underwent amputation. CONCLUSIONS: Owing to the chemosensitive nature of the tumor and possibility of spontaneous regression, neoadjuvant chemotherapy should be considered to prevent extensive or mutilating surgery. PMID- 21617709 TI - From clinical trials to clinical practice: predictors of response to erlotinib in advanced non-small cell lung cancer patients pretreated with chemotherapy. AB - BACKGROUND: Inhibition of the epidermal growth factor receptor pathway with tyrosine kinase inhibitors can improve outcome of patients with advanced non small cell lung cancer after first-line chemotherapy. The use of clinical characteristics and molecular markers may permit the identification of patients who are more likely to benefit from erlotinib. PATIENTS AND METHODS: Retrospective analysis of unselected patients with metastatic non-small cell lung cancer who had previously failed on at least one line of chemotherapy and treated at our institution with erlotinib (150 mg/day orally) until disease progression. Mutations of epidermal growth factor receptor (exon 19-21) and KRAS (codon 12-13) genes were screened with high-resolution melting analysis and identified with direct sequencing. RESULTS: Fifty-three patients were included in the study. The disease control rate was 38%. Median progression-free survival and median overall survival were 4 and 15 months, respectively. Skin rash, diarrhea and mucositis were the most common toxicities of erlotinib. In 19 patients, erlotinib dose was reduced for toxicity. The disease control rate and progression-free survival were significantly better in non-smokers, responders to chemotherapy and patients with epidermal growth factor receptor mutations. Overall survival was longer in patients with skin toxicity and epidermal growth factor receptor mutations. CONCLUSIONS: In our experience, epidermal growth factor receptor mutations, response to previous chemotherapy and non-smoking status were predictors of higher disease control rate and longer progression-free survival. Overall survival was significantly longer in patients with epidermal growth factor receptor mutations and skin toxicity. PMID- 21617711 TI - Patterns of postoperative radiotherapy for head and neck cancer in Italy: a prospective, observational study by the head and neck group of the Italian Association for Radiation Oncology (AIRO). AB - AIMS AND BACKGROUND: Our previous survey showed that the patterns of postoperative radiotherapy (PORT) for head and neck cancer (HNC) in Italy might be suboptimal. A prospective observational study was therefore designed to evaluate this issue in greater detail. METHODS: All radiotherapy centers involved in the HNC Working Group of the Italian Radiation Oncology Association were asked to enter into the study all patients treated with PORT during a 6-month period. RESULTS: A total of 200 patients were accrued by 24 centers from December 2008 to May 2009. Larynx (38%) and oral cavity (34%) were the most common primary sites. The median time between surgery and the start of radiotherapy was 69 days (range, 25-215 days). Seventy-nine percent of cases with no evidence of risk factors for local recurrence were treated with high-dose radiotherapy to the primary site. In about 75% of cases the pN0 neck was included in the target volume. Concomitant chemotherapy was delivered to about 60% of patients with major risk factors and 21% of patients with no risk factors. CONCLUSIONS: Three issues emerged from our study as potential targets for future investigations: the impact on clinical outcome of the interval between surgery and the start of PORT; factors driving radiation oncologists to overtreat volumes at low risk of recurrence; and problems associated with the delivery of concomitant chemotherapy. PMID- 21617712 TI - The INTER-ROMA project--a survey among Italian radiation oncologists on their approach to the treatment of bone metastases. AB - AIMS AND BACKGROUND: Radiotherapy has an established palliative role for bone metastases but despite the large number of patients treated there is still controversy surrounding the optimal radiotherapy schedule to prescribe. The aim of this survey was to determine the decision patterns of Italian radiation oncologists in four different clinical cases of patients with bone metastases. METHODS AND STUDY DESIGN: During the latest national meeting of the Italian Association of Radiation Oncology (AIRO), four clinical cases were presented to attending radiation oncologists. The cases were different with respect to the histology of the primary tumor, performance status, pain before and after analgesics, tumor site, and radiological characteristics of the metastatic lesions. For each clinical case the respondents were asked to give an indication for treatment; prescribe doses, volumes and treatment field arrangements; decide whether to prescribe prophylactic supportive therapy or not; and provide information about factors that particularly influenced prescription. Finally, a descriptive statistical analysis was performed. RESULTS: Three hundred questionnaires were distributed to radiation oncologists attending the congress. One hundred twenty-five questionnaires were returned but only 122 (40.6%) were adequately completed and considered for the analysis. Considerable differences were observed among radiation oncologists in prescribing and delivering radiotherapy for bone metastases. There was also a notable divergence from international guidelines, which will be discussed in this report. CONCLUSIONS: Despite the results of clinical trials, Italian radiation oncologists differ considerably in their decisions on treatment doses and volumes. National guidelines are needed in order that patients can be treated uniformly and better data will become available for evidence-based palliative radiotherapy. PMID- 21617713 TI - AKT signaling pathway in invasive ductal carcinoma of the breast: correlation with ERa, ERbeta and HER-2 expression. AB - AIMS AND BACKGROUND: Estradiol exerts most of its effects by direct binding to the estrogen receptor in breast carcinoma, ERbeta expression is a useful biomarker for breast cancer in a manner that is independent of ERa expression. However, studies evaluating ERbeta expression with certain tumor variables, such as tumor grade and disease-free survival, had produced conflicting results. The Akt signaling pathway currently attracts considerable attention as a new target for effective therapeutic strategies. The current study attempted to compare the relative associations of variables including ERa, ERbeta, HER-2/neu and AKT staining with the presence of metastases or survival. METHODS AND STUDY DESIGN: Immunohistochemical staining was employed to determine the expression of ERa, ERbeta, pAkt and HER-2/neu in 110 cases of primary breast carcinoma. RESULTS: Positive ERa, ERbeta, pAkt and HER-2/neu expressions were respectively observed in 46.4% (51/110), 59.1% (65/110), 40.9% (45/110) and 31.8% (35/110) of the tumors. pAkt was significantly associated with HER-2/neu overexpression (P <0.005) and axillary lymph node metastasis (P <0.05). However, there was no significant relationship between pAkt and ERa, ERbeta, p53 (P >0.05) expressions. Survival analysis showed that pAkt positivity was associated with poor disease free survival of the patients. CONCLUSIONS: The current study suggested that activity of the Akt signaling pathway may indicate a poor prognosis in patients with breast carcinoma. The results implied that estrogen can activate the PI3K Akt pathway through ERa and ERbeta-independent mechanisms in breast cancer. PMID- 21617714 TI - Current role of 111In-DTPA-octreotide scintigraphy in diagnosis of thymic masses. AB - AIMS AND BACKGROUND: Thymic tumors (thymomas and thymic carcinomas) represent 50% of all mediastinal tumors. Thymomas usually express high levels of somatostatin receptors, which enable in vivo imaging with 111In-DTPA-octreotide (OctreoScan(r)). The aim of this study was to further investigate the role of radionuclide techniques in the diagnosis, staging and follow-up of these tumors. METHODS: Eight patients (5 women, 3 men, age range 35-79 years; mean +/- SD 56.1 +/- 15.8 years) entered the study. In 4 patients, myasthenia gravis was the presenting symptom. 111In-DTPA-octreotide scan was performed within 3 weeks after contrast enhanced CT and/or MRI. Planar and tomographic images were acquired within 24 hours of the injection of 111 MBq OctreoScan. The scintigraphic results were defined in correlation with the histological findings. RESULTS: Histology revealed thymoma in 3 patients, thymic carcinoma in 1, insular carcinoma of presumably thymic origin in 1, thymic carcinoid in 1, and thymic hyperplasia in 2 patients. Two thymomas were at stage I, 1 thymoma and 1 thymic carcinoma at stage II, 1 insular carcinoma of presumably thymic origin at stage IV, and 1 thymic carcinoid at stage IV. OctreoScan consistently accumulated in primary and/or metastatic sites of thymic tumors while no radiotracer uptake was detected in the 2 patients with benign thymic hyperplasia. In 1 patient with a very large mediastinal mass (13 cm in largest diameter) and multiple metastatic deposits in the lungs, OctreoScan scintigraphy showed a large area of pathological uptake in the anterior mediastinum and a small area of focal uptake in the cervical-dorsal region of the right lung corresponding to a lymph node expressing somatostatin receptors. CONCLUSIONS: OctreoScan is avidly taken up by thymic tumors, enabling the diagnosis of these tumors and a better evaluation of their extension. It does not accumulate in thymic hyperplasia, thus allowing the differential diagnosis between these 2 pathological conditions. In patients affected by myasthenia gravis, OctreoScan scintigraphy can play an important role in characterizing thymic masses. PMID- 21617715 TI - Interaction effect between the receptor for advanced glycation end products (RAGE) and high-mobility group box-1 (HMGB-1) for the migration of a squamous cell carcinoma cell line. AB - AIMS AND BACKGROUND: The receptor for advanced glycation end products (RAGE) is a multiligand cell surface receptor of the immunoglobulin superfamily and a newly recognized invasion-related gene. High mobility group box-1 (HMGB-1) is a 30-kD protein binding to RAGE and acting as a transcription-factor-like protein that regulates the expression of several genes. In this study, the interaction effect between RAGE and HMGB-1 on the migration of SCC7 cells was investigated along with the inhibitory effect of the drug nifedipine on this interaction effect. METHODS AND STUDY DESIGN: Ten surgical specimens from patients with squamous cell carcinoma (SCC) of the head and neck and a SCC7 cell line were stained using immunohistochemical and immunocytochemical methods. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect RAGE expression in SCC7 cells; Western blot analysis was used to detect HMGB-1 expression in SCC7 cells. An in vitro migration assay (Boyden chamber migration assay) was used for evaluating the interaction effect between RAGE and HMGB-1 on the migration of SCC7 cells. HMGB-1 and various concentrations of nifedipine were tested for their effect on SCC7 cell migration with in vitro migration assays. RESULTS AND CONCLUSIONS: RAGE and HMGB-1 were expressed in almost all human head and neck SCC tissues and in SCC7 cells as detected by immunostaining. The migration assay showed that the interaction between RAGE and HMGB-1 increased SCC7 migration activity depending on the level of HMGB-1, and nifedipine inhibited the interaction effect between RAGE and HMGB-1 on SCC7 cells in a dose-dependent manner. The interaction between RAGE and HMGB-1 could be closely associated with metastasis of SCC of the head and neck. Nifedipine may have an inhibitory effect on tumor metastasis. PMID- 21617716 TI - Expression of cystatin C in human esophageal cancer. AB - AIMS AND BACKGROUND: Cystatin C is a member of the cysteine protease inhibitors and its function is to decrease protease activity. A recent study showed that it was aberrantly expressed in many malignant tumors in association with tumor invasion and metastasis. We attempted to detect its expression in esophageal cancer tissues and adjacent reparative normal tissues. METHODS AND DESIGN: Samples of cancers and non-cancerous esophageal tissues were obtained as matched pairs from 30 surgery patients with esophageal cancer and paraffin embedded. The expression of cystatin C in tissues was investigated by immunohistochemistry. Fisher's exact test was used to analyze the relationship between esophageal cancer tissues and adjacent normal tissues. Furthermore, mRNA was extracted, and reverse transcriptase polymerase chain reaction was performed. RESULTS: The intensity of cystatin C immunostaining in tumor tissues was increased compared to that of adjacent normal tissues. mRNA expression of the cystatin C gene was greater in esophageal cancer than in normal tissues (P <0.05). CONCLUSIONS: Our results indicate that cystatin C may play an important role in the pathogenesis and metastasis of esophageal cancer. PMID- 21617717 TI - Absence of paxillin gene mutation in lung cancer and other common solid cancers. AB - AIMS AND BACKGROUND: Mounting evidence indicates that deregulated cell adhesion is involved in the mechanisms of cancer pathogenesis. A recent study showed that the paxillin gene (PXN) encoding a focal adhesion protein was somatically mutated in lung cancers. The aim of this study was to confirm the presence of PXN mutations in lung cancers as well as in other common solid cancers. METHODS: We analyzed somatic PXN mutations in 45 lung, 45 gastric, 45 colorectal, 45 breast, 45 liver and 45 prostate cancers by polymerase chain reaction and single-strand conformation polymorphism assay. RESULTS: Neither lung nor other cancers were found to be associated with somatic mutations of PXN. CONCLUSIONS: In contrast to the previous report, our study revealed that PXN mutation was absent in lung cancers and other common solid cancers, suggesting that PXN mutation may not play a principal role in solid cancer development. PMID- 21617718 TI - Hereditary paraganglioma syndrome associated with SDHD gene mutations: a patient with multicentric presentation treated with radiotherapy. Case report. AB - INTRODUCTION: Extra-adrenal paragangliomas are rare tumors arising from neuroendocrine cells. Sporadic and hereditary forms have been recognized. Among the latter, the PGL1 and PGL4 syndromes are associated with a high risk of multiple localizations. The treatment of choice for paragangliomas is surgical resection, but in some cases surgery can be difficult due to particular or multiple tumor sites or may result in severe neurological deficits. In such cases radiotherapy can be an effective alternative. In this paper we describe the case of a patient affected by hereditary paraganglioma syndrome with multicentric presentation who was treated at our center by external radiotherapy. CASE REPORT: A 55-year-old man presented in April 2008 with multiple paragangliomas: one in the left pontocerebellar angle, two in the middle neck, one mediastinal paraaortic mass, and an abdominal paraaortic lesion. The left pontocerebellar and mediastinal tumors were treated with three-dimensional conformal radiotherapy (3D CRT) at total doses of 50.40 Gy and 55.80 Gy, respectively. The neck lesions were treated with intensity-modulated radiotherapy (IMRT) at a total dose of 55.80 Gy. The abdominal paraaortic lesion was surgically resected. RESULTS: No severe acute or late toxicity as evaluated with the EORTC-RTOG scale was observed. Fourteen months after the end of radiotherapy a whole body CT scan showed that the tumor lesions were stable in size and in their relations to contiguous structures. The arterial pressure was controlled by medical therapy and urine catecholamine levels were within the normal range. CONCLUSIONS: We believe that in patients affected by unresectable paragangliomas radiotherapy is a safe and effective alternative to surgery. The use of high-dose conformity techniques such as 3D-CRT and IMRT will allow higher local control rates with relatively few side effects thanks to the possibility of dose escalation and reduction of the amount of irradiated healthy tissues. PMID- 21617719 TI - Megavoltage CT images of helical tomotherapy unit for radiation treatment simulation: impact on feasibility of treatment planning in a prostate cancer patient with bilateral femoral prostheses. AB - Metal prosthesis artefacts on CT images can be a significant problem in the definition of volumes of interest, dose calculation and patient setup in modern radiotherapy. We experienced considerable difficulties in defining the organs at risk and treatment volumes on kVCT images of standard CT simulation in a prostate cancer patient due to the presence of bilateral femoral prostheses causing artefacts. As shown in the current case, MVCT images of the patient in the treatment position obtained using a helical tomotherapy unit can provide sufficient morphological information to define the pelvic anatomic structures for radical prostate treatment planning. The patient completed the planned treatment and at 90 days after the end of treatment no severe side effects were recorded. Since there have been few reports on the use of MVCT images to overcome the problem of hip prosthesis artefacts, a brief literature review was also carried out. PMID- 21617720 TI - Multidisciplinary approach in the treatment of malignant paraganglioma of the glomus vagale: a case report. AB - Malignant paraganglioma of the glomus vagale is a rare tumor entity originating from paraganglia or glomus cells. It typically affects middle age. It differs from benign paraganglioma because of its rapid growth and more aggressive clinical behavior. We report the case of a 40-year-old man presenting with a 5 cm lesion in the upper right cervical region detected by computed tomography (CT) and magnetic resonance imaging (MRI), which also showed enlargement of ipsilateral spinal and jugulodigastric lymph nodes with contrast enhancement. Clinical manifestations at diagnosis included a partial neurological deficit involving the right cranial nerves X, XI and XII. Tumor vascularization was assessed by digital angiography. The tumor mass was entirely removed by a right cervical approach with en-bloc resection with the regional lymph nodes. Histopathological examination showed a paraganglioma with cellular pleomorphism, necrotic microfoci, perineural infiltration and angiogenesis. Massive metastases in two of three jugulodigastric and one spinal lymph nodes on the right side were also detected. Postoperative workup included MRI, positron emission tomography (PET)/CT, meta-iodine-benzyl-guanidine (MIBG) scan, liver ultrasound and chest radiography. Subsequently, the patient underwent conformal radiotherapy with concomitant cisplatin administration. At the last clinical and radiological follow-up examination 5 years after completion of treatment, the patient was free of tumor recurrence. The integrated treatment by surgery and chemoradiation was feasible and effective in the management of this rare case of malignant paraganglioma of the glomus vagale. Multicenter studies should be done to increase the knowledge of tumor presentation and natural history and to analyze the possible treatment options. PMID- 21617721 TI - A case report of thyroid gland metastasis associated with lung metastasis from colon cancer. AB - Thyroid gland metastasis of malignant tumors is observed in 1.9% to 9.5% of histologically examined autopsy cases. Thyroid metastasis from colon cancer is extremely rare and the prognosis is poor. Here we report a case of lung metastasis and thyroid gland metastasis following sigmoid colon cancer surgery. In 2000, a 58-year-old woman underwent a sigmoid colectomy for sigmoid colon cancer. In 2005, a metastatic lung tumor was detected by chest CT. The patient underwent a partial thoracoscopic resection of the left lung in April 2005. On a CT scan taken 3 years and 4 months after the lung resection, a tumor mass was observed in the left lung and a low-absorption region with an unclear border was seen in the left lobe of the thyroid gland. Thyroid aspiration cytology showed adenocarcinoma, and a diagnosis of thyroid gland metastasis from sigmoid colon cancer was made. In April 2008 a subtotal thyroidectomy was performed. Following surgery, the patient underwent chemotherapy with mFOLFOX6 and bevacizumab. Nevertheless a number of lung metastases and expressions of lung metastasis were subsequently observed. Histopathological examination revealed a number of metastases of differentiated papillary adenocarcinoma in the thyroid gland from colon cancer. PMID- 21617722 TI - Primary breast lymphoma clinically mimicking acute mastitis: a case report. AB - Extranodal non-Hodgkin lymphoma of the breast is a rare disease. We present a case of primary breast lymphoma with atypical clinical manifestations that looked like acute mastitis. A 46-year-old woman had noted a painful swelling in the right breast for 2 months. The mass had an inflammatory appearance and acute mastitis was the clinical impression. She underwent a core biopsy of the mass, and pathology showed inflammatory changes. The inflammatory mass regressed and recurred during hospitalization, and further incision with debridement was done. The histological findings were consistent with diffuse large B cell lymphoma. Systemic examination found stage IIE disease. She received chemotherapy and local radiation is planned. PMID- 21617723 TI - Squamous cell carcinoma of the umbilicus: management of an unusual localization. AB - We describe the case of a squamous cell carcinoma spreading to the skin and regional lymph nodes from the umbilicus. Bilateral inguinal lymphadenectomy and a session of electrochemotherapy with bleomycin 15 mg/m2 were performed. However, because of the development of new cutaneous nodules in the abdominopelvic region, we performed targeted palliative therapy with erlotinib 150 mg/day. Targeted adjuvant therapy was preferred to the use of a major cytotoxic agent because of the high risk of superinfection and heart failure. Erlotinib produced a partial clinical response with reduction of the number and size of the skin nodules. CT scan performed after 60 days of treatment did not show any new lesions. To our knowledge, this is the first report of an umbilical metastatic squamous cell carcinoma treated with modern targeted therapy. This therapeutic strategy can be considered a valid palliative option in the management of metastatic cutaneous nodules of this rare primary site. PMID- 21617724 TI - Successful treatment with GEMOX in patient with metastatic pancreatic adenosquamous carcinoma. AB - BACKGROUND: Pancreatic adenosquamous carcinoma (PASC) is a rare subtype of pancreatic cancer characterized by a dual histological component and aggressive behavior. This form is not well known, its histogenesis is uncertain, and there are different opinions on the diagnostic histopathological criteria. The differential diagnosis with more common ductal adenocarcinoma, squamous cell carcinoma, squamous or adenosquamous metastases is complex. The available therapies do not improve the poor prognosis and it is difficult to find long-term survivors (more than 1 year), even after demolitive surgery with complementary therapies. CASE REPORT: We report a case of advanced PASC with excellent progression-free survival and overall survival, 20 months and 29 months, respectively. Furthermore, an almost complete response was obtained to first-line chemotherapy with gemcitabine/oxaliplatin (GEMOX) followed by maintenance gemcitabine. CONCLUSION: GEMOX followed by gemcitabine as maintenance could be an effective treatment for this pancreatic entity. Further reports are needed to confirm this outcome. PMID- 21617725 TI - Concurrence of UGT1A polymorphism and end-stage renal disease leads to severe toxicities of irinotecan in a patient with metastatic colon cancer. AB - AIMS AND BACKGROUND: Colorectal cancer is one of the most common malignancies in the world, and irinotecan (CPT-11) is useful in its treatment. However, the safety and pharmacokinetics of irinotecan in dialysis patients with metastatic colorectal cancer are unclear. CASE REPORT: We report the case of a 74-year-old man receiving chronic hemodialysis who had metastatic colorectal cancer. Palliative chemotherapy with irinotecan (80 mg/m2 weekly) was administered after hemodialysis. Blood samples were collected before and 1.5, 3, 6, 9, and 15 hours after administration of irinotecan. The peak serum concentrations (Cmax) of irinotecan and SN-38 in this patient were 1,480 and 17.8 ng/mL, respectively, which were similar to the reported values in patients with normal renal function after a similar dose of irinotecan (75 mg/m2). The area under the serum concentration-time curve (AUC0-infinity) was 8,240 ng*h/mL for irinotecan and 619 ng*h/mL for SN-38. The AUC0-infinity for SN-38 was markedly higher than that for patients with normal renal function. Sequencing analysis of the UGT1A genes found that the patient had variant alleles of UGT1A1*28, UGT1A1*60 and UGT1A9*22, which may lead to decreased glucuronidation and excretion of SN-38, and may account for increased irinotecan-related toxicity. The patient developed febrile grade 4 neutropenia on day 7 after chemotherapy and died of septic shock on day 14. CONCLUSIONS: UGT1A polymorphisms and renal failure may lead to accumulation of SN 38, which may have played a role in the death of this patient. Irinotecan should be used cautiously in dialysis patients with metastatic colorectal cancer and screening for UGT1A polymorphisms may help in identifying patients with lower SN 38 glucuronidation rates and greater susceptibility to irinotecan-induced toxicity. PMID- 21617726 TI - Distribution of coding apoptotic gene polymorphisms in women with extreme phenotypes of breast cancer predisposition and tolerance. AB - AIMS AND BACKGROUND: Comparison of subjects with extreme phenotypes of cancer susceptibility and tolerance allows to detect low-penetrance gene-disease interactions with a relatively small study size. METHODS AND STUDY DESIGN: We analyzed the distribution of 19 coding apoptotic gene polymorphisms (Bid Gly10Ser; Casp2 Leu141Val; Casp5 Ala90Thr and Val318Leu; Casp7 Glu255Asp; Casp8 His302Asp; Casp9 Val28Ala, His173Arg and Arg221Gln; Casp10 Ile479Leu; Faim Thr117Ala and Ser127Leu; DR4 Arg141His, Thr209Arg, Ala228Glu and Lys441Arg; Survivin Lys129Glu; TNFR1 Gln121Arg; XIAP Pro423Gln) in 121 breast cancer patients with clinical features of a hereditary predisposition (family history and/or early onset and/or bilaterality) and 142 elderly tumor-free women. RESULTS: None of the individual single nucleotide polymorphisms (SNPs) demonstrated an association with breast cancer risk. The analysis of gene interactions revealed that the combination of XIAP Pro423Gln (rs5956583) AA genotype with Casp7 Glu255Asp (rs2227310) CG genotype appeared to prevail in "supercases" relative to "supercontrols" (25/121 [21%] vs 11/142 [8%], P = 0.002). We attempted to validate this association in the second round of case control analysis, which involved 519 randomly selected breast cancer patients and 509 age-matched healthy women, but no difference was detected upon this comparison. CONCLUSIONS: Coding apoptotic gene polymorphisms do not play a major role in BC predisposition. The results of this investigation may be considered while designing future studies on breast cancer-associated candidate SNPs. PMID- 21617727 TI - Trabectedin-related rhabdomyolysis: an uncommon but fatal toxicity. AB - Rhabdomyolysis is defined as the dissolution of striped muscle characterized by the leakage of intracellular muscle components into the circulation, which can ultimately lead to renal failure with a possible fatal outcome. Trabectedin is a potential cause of rhabdomyolysis. Herein, we describe a case of rhabdomyolysis in a female patient with recurrent metastatic leiomyosarcoma of the uterus and who had full recovery, and we review the already published cases in order to identify a common pattern of emergence. PMID- 21617728 TI - Problems in dealing with very rare adverse effects of new anticancer drugs: the example of trabectedin. PMID- 21617729 TI - Does in situ melanoma really come before invasive melanoma? Descriptive epidemiology questions this relationship. PMID- 21617730 TI - Spike rate and spike timing contributions to coding taste quality information in rat periphery. AB - There is emerging evidence that individual sensory neurons in the rodent brain rely on temporal features of the discharge pattern to code differences in taste quality information. In contrast, investigations of individual sensory neurons in the periphery have focused on analysis of spike rate and mostly disregarded spike timing as a taste quality coding mechanism. The purpose of this work was to determine the contribution of spike timing to taste quality coding by rat geniculate ganglion neurons using computational methods that have been applied successfully in other systems. We recorded the discharge patterns of narrowly tuned and broadly tuned neurons in the rat geniculate ganglion to representatives of the five basic taste qualities. We used mutual information to determine significant responses and the van Rossum metric to characterize their temporal features. While our findings show that spike timing contributes a significant part of the message, spike rate contributes the largest portion of the message relayed by afferent neurons from rat fungiform taste buds to the brain. Thus, spike rate and spike timing together are more effective than spike rate alone in coding stimulus quality information to a single basic taste in the periphery for both narrowly tuned specialist and broadly tuned generalist neurons. PMID- 21617731 TI - Disinhibition bursting of dopaminergic neurons. AB - Substantia nigra pars compacta (SNpc) dopaminergic neurons receive strong tonic inputs from GABAergic neurons in the substantia nigra pars reticulata (SNpr) and globus pallidus (GP), and glutamatergic neurons in the subthalamic nucleus. The presence of these tonic inputs raises the possibility that phasic disinhibition may trigger phasic bursts in dopaminergic neurons. We first applied constant NMDA and GABA(A) conductances onto a two-compartment single cell model of the dopaminergic neuron (Kuznetsov et al., 2006). The model exhibited disinhibition bursting upon stepwise removal of inhibition. A further bifurcation analysis suggests that disinhibition may be more robust than excitation alone in that for most levels of NMDA conductance, the cell remains capable of bursting even after a complete removal of inhibition, whereas too much excitatory input will drive the cell into depolarization block. To investigate the network dynamics of disinhibition, we used a modified version of an integrate-and-fire based model of the basal ganglia (Humphries et al., 2006). Synaptic activity generated in the network was delivered to the two-compartment single cell dopaminergic neuron. Phasic activation of the D1-expressing medium spiny neurons in the striatum (D1STR) produced disinhibition bursts in dopaminergic neurons through the direct pathway (D1STR to SNpr to SNpc). Anatomical studies have shown that D1STR neurons have collaterals that terminate in GP. Adding these collaterals to the model, we found that striatal activation increased the intra-burst firing frequency of the disinhibition burst as the weight of this connection was increased. Our studies suggest that striatal activation is a robust means by which disinhibition bursts can be generated by SNpc dopaminergic neurons, and that recruitment of the indirect pathway via collaterals may enhance disinhibition bursting. PMID- 21617732 TI - Spike correlations - what can they tell about synchrony? AB - Sensory and cognitive processing relies on the concerted activity of large populations of neurons. The advent of modern experimental techniques like two photon population calcium imaging makes it possible to monitor the spiking activity of multiple neurons as they are participating in specific cognitive tasks. The development of appropriate theoretical tools to quantify and interpret the spiking activity of multiple neurons, however, is still in its infancy. One of the simplest and widely used measures of correlated activity is the pairwise correlation coefficient. While spike correlation coefficients are easy to compute using the available numerical toolboxes, it has remained largely an open question whether they are indeed a reliable measure of synchrony. Surprisingly, despite the intense use of correlation coefficients in the design of synthetic spike trains, the construction of population models and the assessment of the synchrony level in live neuronal networks very little was known about their computational properties. We showed that many features of pairwise spike correlations can be studied analytically in a tractable threshold model. Importantly, we demonstrated that under some circumstances the correlation coefficients can vanish, even though input and also pairwise spike cross correlations are present. This finding suggests that the most popular and frequently used measures can, by design, fail to capture the neuronal synchrony. PMID- 21617733 TI - Deep brain stimulation in the media: over-optimistic portrayals call for a new strategy involving journalists and scientists in ethical debates. AB - Deep brain stimulation (DBS) is optimistically portrayed in contemporary media. This already happened with psychosurgery during the first half of the twentieth century. The tendency of popular media to hype the benefits of DBS therapies, without equally highlighting risks, fosters public expectations also due to the lack of ethical analysis in the scientific literature. Media are not expected (and often not prepared) to raise the ethical issues which remain unaddressed by the scientific community. To obtain a more objective portrayal of DBS in the media, a deeper collaboration between the science community and journalists, and particularly specialized ones, must be promoted. Access to databases and articles, directly or through science media centers, has also been proven effective in increasing the quality of reporting. This article has three main objectives. Firstly, to explore the past media coverage of leukotomy, and to examine its widespread acceptance and the neglect of ethical issues in its depiction. Secondly, to describe how current enthusiastic coverage of DBS causes excessive optimism and neglect of ethical issues in patients. Thirdly, to discuss communication models and strategies to enhance media and science responsibility. PMID- 21617734 TI - Auditory perception of self-similarity in water sounds. AB - Many natural signals, including environmental sounds, exhibit scale-invariant statistics: their structure is repeated at multiple scales. Such scale-invariance has been identified separately across spectral and temporal correlations of natural sounds (Clarke and Voss, 1975; Attias and Schreiner, 1997; Escabi et al., 2003; Singh and Theunissen, 2003). Yet the role of scale-invariance across overall spectro-temporal structure of the sound has not been explored directly in auditory perception. Here, we identify that the acoustic waveform from the recording of running water is a self-similar fractal, exhibiting scale-invariance not only within spectral channels, but also across the full spectral bandwidth. The auditory perception of the water sound did not change with its scale. We tested the role of scale-invariance in perception by using an artificial sound, which could be rendered scale-invariant. We generated a random chirp stimulus: an auditory signal controlled by two parameters, Q, controlling the relative, and r, controlling the absolute, temporal structure of the sound. Imposing scale invariant statistics on the artificial sound was required for its perception as natural and water-like. Further, Q had to be restricted to a specific range for the sound to be perceived as natural. To detect self-similarity in the water sound, and identify Q, the auditory system needs to process the temporal dynamics of the waveform across spectral bands in terms of the number of cycles, rather than absolute timing. We propose a two-stage neural model implementing this computation. This computation may be carried out by circuits of neurons in the auditory cortex. The set of auditory stimuli developed in this study are particularly suitable for measurements of response properties of neurons in the auditory pathway, allowing for quantification of the effects of varying the statistics of the spectro-temporal statistical structure of the stimulus. PMID- 21617735 TI - Dissecting the contribution of individual receptor subunits to the enhancement of N-methyl-d-aspartate currents by dopamine D1 receptor activation in striatum. AB - Dopamine, via activation of D1 receptors, enhances N-methyl-d-aspartate (NMDA) receptor-mediated responses in striatal medium-sized spiny neurons. However, the role of specific NMDA receptor subunits in this enhancement remains unknown. Here we used genetic and pharmacological tools to dissect the contribution of NR1 and NR2A/B subunits to NMDA responses and their modulation by dopamine receptors. We demonstrate that D1 enhancement of NMDA responses does not occur or is significantly reduced in mice with genetic knock-down of NR1 subunits, indicating a critical role of these subunits. Interestingly, spontaneous and evoked alpha amino-3-hydroxyl-5-methyl-4-isoxazole-propionic acid (AMPA) receptor-mediated responses were significantly enhanced in NR1 knock-down animals, probably as a compensatory mechanism for the marked reduction in NMDA receptor function. The NMDA receptor subunits NR2A and NR2B played differential roles in D1 modulation. Whereas genetic deletion or pharmacological blockade of NR2A subunits enhanced D1 potentiation of NMDA responses, blockade of NR2B subunits reduced this potentiation, suggesting that these regulatory subunits of the NMDA receptor counterbalance their respective functions. In addition, using D1 and D2 receptor EGFP-expressing mice, we demonstrate that NR2A subunits contribute more to NMDA responses in D1-MSSNs, whereas NR2B subunits contribute more to NMDA responses in D2 cells. The differential contribution of discrete receptor subunits to NMDA responses and dopamine modulation in the striatum has important implications for synaptic plasticity and selective neuronal vulnerability in disease states. PMID- 21617736 TI - Determining the nature of prefrontal cortex recruitment after traumatic brain injury: a response to Turner. PMID- 21617737 TI - Soil Physical Constraints on Intrinsic Biodegradation of Petroleum Vapors in a Layered Subsurface. AB - Naturally occurring biodegradation of petroleum hydrocarbons in the vadose zone depends on the physical soil environment influencing field-scale gas exchange and pore-scale microbial metabolism. In this study, we evaluated the effect of soil physical heterogeneity on biodegradation of petroleum vapors in a 16-m-deep, layered vadose zone. Soil slurry experiments (soil/water ratio 10:30 w/w, 25 degrees C) on benzene biodegradation under aerobic and well-mixed conditions indicated that the biodegradation potential in different textured soil samples was related to soil type rather than depth, in the order: sandy loam > fine sand > limestone. Similarly, O(2) consumption rates during in situ respiration tests performed at the site were higher in the sandy loam than in the fine sand, although the difference was less significant than in the slurries. Laboratory and field data generally agreed well and suggested a significant potential for aerobic biodegradation, even with nutrient-poor and deep subsurface conditions. In slurries of the sandy loam, the biodegradation potential declined with increasing in situ water saturation (i.e., decreasing air-filled porosity in the field). This showed a relation between antecedent undisturbed field conditions and the slurry biodegradation potential, and suggested airfilled porosity to be a key factor for the intrinsic biodegradation potential in the field. PMID- 21617738 TI - RadBall Technology Testing in the Savannah River Site's Health Physics Instrument Calibration Laboratory. AB - The United Kingdom's National Nuclear Laboratory (NNL) has developed a radiation mapping device that can locate and quantify radioactive hazards within contaminated areas of the nuclear industry. The device, known as RadBall(TM), consists of a colander-like outer collimator that houses a radiation-sensitive polymer sphere. The collimator has over two hundred small holes; thus, specific areas of the polymer sphere are exposed to radiation becoming increasingly more opaque in proportion to the absorbed dose. The polymer sphere is imaged in an optical-CT scanner that produces a high resolution 3D map of optical attenuation coefficients. Subsequent analysis of the optical attenuation data provides information on the spatial distribution of sources in a given area forming a 3D characterization of the area of interest. The RadBall(TM) technology has been deployed in a number of technology trials in nuclear waste reprocessing plants at Sellafield in the United Kingdom and facilities of the Savannah River National Laboratory (SRNL). This paper summarizes the tests completed at SRNL Health Physics Instrument Calibration Laboratory (HPICL). PMID- 21617739 TI - Predicting Air Quality in Smart Environments. AB - The pervasive sensing technologies found in smart environments offer unprecedented opportunities for monitoring and assisting the individuals who live and work in these spaces. As aspect of daily life that is often overlooked in maintaining a healthy lifestyle is the air quality of the environment. In this paper we investigate the use of machine learning technologies to predict CO(2) levels as an indicator of air quality in smart environments. We introduce techniques for collecting and analyzing sensor information in smart environments and analyze the correlation between resident activities and air quality levels. The effectiveness of our techniques is evaluated using three physical smart environment testbeds. PMID- 21617740 TI - RadBall Technology Testing and MCNP Modeling of the Tungsten Collimator. AB - The United Kingdom's National Nuclear Laboratory (NNL) has developed a remote, non-electrical, radiation-mapping device known as RadBall(TM), which can locate and quantify radioactive hazards within contaminated areas of the nuclear industry. RadBall(TM) consists of a colander-like outer shell that houses a radiation-sensitive polymer sphere. The outer shell works to collimate radiation sources and those areas of the polymer sphere that are exposed react, becoming increasingly more opaque, in proportion to the absorbed dose. The polymer sphere is imaged in an optical-CT scanner, which produces a high resolution 3D map of optical attenuation coefficients. Subsequent analysis of the optical attenuation matrix provides information on the spatial distribution of sources in a given area forming a 3D characterization of the area of interest. RadBall(TM) has no power requirements and can be positioned in tight or hard-to reach locations. The RadBall(TM) technology has been deployed in a number of technology trials in nuclear waste reprocessing plants at Sellafield in the United Kingdom and facilities of the Savannah River National Laboratory (SRNL). This study focuses on the RadBall(TM) testing and modeling accomplished at SRNL. PMID- 21617741 TI - Silicon-Neuron Design: A Dynamical Systems Approach. AB - We present an approach to design spiking silicon neurons based on dynamical systems theory. Dynamical systems theory aids in choosing the appropriate level of abstraction, prescribing a neuron model with the desired dynamics while maintaining simplicity. Further, we provide a procedure to transform the prescribed equations into subthreshold current-mode circuits. We present a circuit design example, a positive-feedback integrate-and-fire neuron, fabricated in 0.25 MUm CMOS. We analyze and characterize the circuit, and demonstrate that it can be configured to exhibit desired behaviors, including spike-frequency adaptation and two forms of bursting. PMID- 21617742 TI - Discovering Activities to Recognize and Track in a Smart Environment. AB - The machine learning and pervasive sensing technologies found in smart homes offer unprecedented opportunities for providing health monitoring and assistance to individuals experiencing difficulties living independently at home. In order to monitor the functional health of smart home residents, we need to design technologies that recognize and track activities that people normally perform as part of their daily routines. Although approaches do exist for recognizing activities, the approaches are applied to activities that have been pre-selected and for which labeled training data is available. In contrast, we introduce an automated approach to activity tracking that identifies frequent activities that naturally occur in an individual's routine. With this capability we can then track the occurrence of regular activities to monitor functional health and to detect changes in an individual's patterns and lifestyle. In this paper we describe our activity mining and tracking approach and validate our algorithms on data collected in physical smart environments. PMID- 21617743 TI - Preliminary Investigation of the Dosimetric Properties of 'RadGel' AB - A preliminary investigation into the efficacy of a new 3D dosimetry material, RadGelTM, for verification of radiation therapy dose distributions is presented. Small volumes of RadGelTM were found to exhibit a linear, reproducible response to dose. A gradual increase in optical-density (OD) with time was observed, suggesting scanning should be completed within 18 hours to keep a linear correlation of R(2) > 0.99. A larger 10 cm diameter volume of RadGelTM was irradiated with a rotationally symmetric "spoke" plan designed to rigorously evaluate scanner/dosimeter combined performance. The dosimeter was imaged with the Duke Mid-sized Optical-CT Scanner (DMOS). Promising OD and corresponding dose maps were obtained. Edge artefacts were observed and are suspected to be exacerbated by the particular container used in this early study. Further studies will evaluate new containers and methods for refractive matching at the gel container-fluid interface. PMID- 21617744 TI - ScanSim: A tool for simulating optical-CT imaging. AB - A software tool has been developed that can simulate image formation in a variety of optical-CT scanning configurations. The formalism of the simulation is introduced, including two main modes: a diverging point source mode, and a converging broad beam mode. Preliminary results are presented for scanning Presage dosimeters in both modes and immersed in refractive media of widely varying refractive index (RI), including air, water, and a fully matched medium. Pronounced differences in the edge artifacts and accuracy of reconstructed coefficients is observed. The ScanSim software is shown to be a useful tool to investigate and quantify many aspects of optical-CT image formation, including reducing dependence on matching fluids. PMID- 21617745 TI - Dosimetry challenges for implementing emerging technologies. AB - During the last 10 years, radiation therapy technologies have gone through major changes, mainly related introduction of sophisticated delivery and imaging techniques to improve the target localization accuracy and dose conformity. While implementation of these emerging technologies such as image-guided SRS/SBRT, IMRT/IMAT, IGRT, 4D motion management, and special delivery technologies showed substantial clinical gains for patient care, many other factors, such as training/quality, efficiency/efficacy, and cost/effectiveness etc. remain to be challenging. This talk will address technical challenges for dosimetry verification of implementing these emerging technologies in radiation therapy. PMID- 21617746 TI - Enhancing the Cultural Relevance of Empirically-Supported Mental Health Interventions. AB - Evidence-based practice (EBP) has become a hot topic in clinical social work and other mental health disciplines. Mental health professionals have called attention to the need for clinical decision-making to be based on the best available empirically supported treatments integrated with client preferences, values, and circumstances. This movement has greatly stimulated mental health professionals to develop, test, and adopt efficacious treatments for clients with psychological problems, but what is missing in the literature is the cultural context in which these treatments must be implemented to be effective with racial/ethnic minority populations. Herein, we utilize the culturally centered framework of Bernal, Bonilla and Bellido (1995) to examine its utility in assessing to what extent empirically supported mental health treatments incorporate culturally relevant components. PMID- 21617747 TI - Color Perception of 3D Objects: Constancy with Respect To Variation of Surface Gloss. AB - What determines the color appearance of real objects viewed under natural conditions? The light reflected from different locations on a single object can vary enormously. This variation is enhanced when the material properties of the object are changed from matte to glossy. Yet humans have no trouble assigning a color name to most things. We studied how people perceive the color of spheres in complex scenes. Observers viewed graphics simulations of a three-dimensional scene containing two spheres, test and match. The observer's task was to adjust the match sphere until its color appearance was the same as that of the test sphere. The match sphere was always matte, and observers varied its color by changing the simulated spectral reflectance function. The surface gloss of the test spheres was varied across conditions. The data show that for fixed test sphere body reflectance, color appearance depends on surface gloss. This effect is small, however, compared to the variation that would be expected if observers simply matched the average of the light reflected from the test. PMID- 21617748 TI - Asymmetric phenotype of Axenfeld-Rieger anomaly and aniridia associated with a novel PITX2 mutation. AB - PURPOSE: To evaluate the asymmetry of the anterior segment phenotype between the two eyes of a patient with Axenfeld-Rieger syndrome (ARS). METHODS: The entire database of a tertiary glaucoma practice was screened for patients with ARS. The medical records of patients with ARS were reviewed. The clinical characteristics of ocular examination of the two eyes of each patient were recorded and compared. Dental and medical information were also reviewed where available. The anterior segment phenotype was tabulated to assess asymmetry. Asymmetric anterior segment characteristics of patients with ARS were compared with reported cases in the literature. RESULTS: Eight patients with ARS were identified from screening of more than 5,000 patients of a tertiary glaucoma practice. All patients had Axenfeld-Rieger anomaly in both eyes except one patient presented with an asymmetric phenotype of the anterior segment with features of Axenfeld-Rieger anomaly in one eye, but aniridia in the other eye. This patient had non-ocular findings including flat midface, hypodontia with lack of an upper incisor, and redundant periumbilical skin, typical for ARS. A heterozygous C>T nucleotide substitution was identified in exon 4 of the pituitary homeobox 2 (PITX2) gene, resulting in the replacement of a glutamine codon (CAG) with a stop codon (TAG) at amino acid position 67. This mutation is denoted c.199C>T at the cDNA level or p.Gln67Stop (or Q67X) at the protein level. Only three cases with asymmetric anterior segment phenotype between the two eyes of a patient with AGS have been reported in the literature. CONCLUSIONS: Variability in phenotype may occur between the two eyes of an individual affected by ARS. The current case undermines the advantage of genetic testing to correctly diagnose a rare disease. PMID- 21617749 TI - Analysis on the alterations of lens proteins by Vitex negundo in selenite cataract models. AB - PURPOSE: Cataract is the leading cause of blindness and is associated with oxidative damage and protein modification in the lens. In the present study, we have employed proteomic and microscopic approaches to investigate the attenuation of selenite cataract by the flavonoids from Vitex negundo (FVN). METHODS: To demonstrate this attenuation, Sprague-Dawley rat pups were divided into control (G I), selenite induced (G II), and selenite + FVN treated (G III). Cataract was induced by single subcutaneous injection of sodium selenite (4 mg/Kg bodyweight) on the 10th day and FVN (1 mg/Kg bodyweight) administered intraperitoneally from the 8th to the 15th day. RESULTS: Our study indicated that chaperone property of alpha-crystallin and soluble protein levels were reduced in the selenite induced group. Post translational modifications identified by two dimensional polyacrylamide gel electrophoresis (2D-PAGE) and immunoblot analysis revealed the loss of cytoskeletal proteins in selenite induced group. Damage of lenticular membrane and abnormal fiber structure were observed by electron microscopy. CONCLUSIONS: The results of this study suggest that FVN modulated selenite induced cataractogensis in rat pups by preventing loss of chaperone property, various changes in lens proteins, and lens structure, further strengthening its protective role. PMID- 21617750 TI - XRCC1 and XPD DNA repair gene polymorphisms: a potential risk factor for glaucoma in the Pakistani population. AB - PURPOSE: The present study was designed to determine the association of polymorphisms of the DNA repair genes X-ray cross-complementing group 1 (XRCC1) (c.1316G>A [rs25487]) and xeroderma pigmentosum complementation group D (XPD) (c.2298A>C [rs13181]) with primary open-angle glaucoma (POAG) and primary closed angle glaucoma (PCAG). METHODS: In this prospective case-control study, polymerase chain reaction-restriction fragment length polymorphism analysis was used to study the association of XRCC1 and XPD with 160 POAG patients, 163 PCAG patients, and 193 unaffected controls. RESULTS: XRCC1 rs25487 was found to be significantly associated specifically with male POAG patients (chi(2) = 13.2 [p = 0.001]), only for the dominant model (odds ratio [OR] = 2.65 [95% confidence interval [CI] = 1.44-4.85], p < 0.005). In addition XPD rs13181 was also found to be associated with male POAG patients (chi(2) = 12.1 [p < 0.005]), for both dominant (OR = 2.44 [95% CI = 1.33-4.47], p < 0.005) as well as recessive model (OR = 3.62 [95% CI = 1.45-9.01], p < 0.01). Combined genotypes of both the genes revealed that the heterozygote AC/GA was significantly associated with the male POAG patients (z = 3.00 [p < 0.001]). The AA/GG genotype was present at a higher frequency in the male controls and the AA/GA in the female controls and could thus have a protective role in males and females, respectively. CONCLUSIONS: We postulate that defects in the DNA repair genes XRCC1 and XPD may possibly be associated with the progression of POAG in male patients of Pakistani origin. PMID- 21617751 TI - Genotype-phenotype correlations of TGFBI p.Leu509Pro, p.Leu509Arg, p.Val613Gly, and the allelic association of p.Met502Val-p.Arg555Gln mutations. AB - PURPOSE: Investigate the genotype-phenotype correlations for five TGFBI (transforming growth factor, beta-induced) mutations including one novel pathogenic variant and one complex allele affecting the fourth FAS1 domain of keratoepithelin, and their potential effects on the protein's structure. METHODS: Three unrelated families were clinically diagnosed with lattice corneal dystrophy (CD) and one with an unclassified CD of Bowman's layer. Mutations in the TGFBI gene were detected by direct sequencing, and the functional impact of each variant was predicted using in silico algorithms. Corneal phenotypes, including histological examinations, were compared with the literature data. Furthermore, molecular modeling studies of these mutations were performed. RESULTS: Two distinct missense mutations affecting the same residue at position 509 of keratoepithelin: p.Leu509Pro (c.1526T>C) and p.Leu509Arg (c.1526T>G) were found to be associated with a lattice-type CD. The novel p.Val613Gly (c.1828T>G) TGFBI mutation was found in a sporadic case of an Algerian individual affected by lattice CD. Finally, the Bowman's layer CD was linked to the association in cis of the p.Met502Val and p.Arg555Gln variants, leading to the reclassification of this CD as atypical Thiel-Behnke CD. Structural modeling of these TGFBI mutations argues in favor of these mutations being responsible for instability and/or incorrect folding of keratoepithelin, predictions that are compatible with the clinical diagnoses. CONCLUSIONS: Description of a novel TGFBI mutation and a complex TGFBI allele further extends the mutational spectrum of TGFBI. Moreover, we show convincing evidence that TGFBI mutations affecting Leu509 are linked to the lattice phenotype in two unrelated French families, contrasting with findings previously reported. The p.Leu509Pro was reported to be associated with both amyloid and non-amyloid aggregates, whereas p.Leu509Arg has been described as being responsible for Epithelial Basement Membrane Dystrophy (EBMD). PMID- 21617752 TI - Oxidative stress, mitochondrial dysfunction and calcium overload in human lamina cribrosa cells from glaucoma donors. AB - PURPOSE: Oxidative stress is implicit in the pathological changes associated with glaucoma. The purpose of this study was to compare levels of oxidative stress in glial fibrillary acid-negative protein (GFAP) lamina cribrosa (LC) cells obtained from the optic nerve head (ONH) region of 5 normal (NLC) and 4 glaucomatous (GLC) human donor eyes and to also examine mitochondrial function and calcium homeostasis in this region of the ONH. METHODS: Intracellular reactive oxygen species (ROS) production was examined by a thiobarbituric acid reactive substances (TBARS) assay which measures malondialdehyde (MDA), a naturally occurring product of lipid peroxidation and is used as an indicator of oxidative stress. Mitochondrial membrane potential (MMP) and intracellular calcium ([Ca(2+)](i)) levels were evaluated by flow cytometry using the JC-1 (5,5',6,6' tetrachloro-1,1',3,3'-tetrabenzimidazolecarbocyanine iodide) and fluo-4/AM probes respectively. Anti-oxidant and Ca(2+) transport system gene and protein expression were determined by real time polymerase chain reaction (RT-PCR) using gene-specific primer/probe sets and western immunoblotting, respectively. RESULTS: Intracellular ROS production was increased in GLC compared to NLC (27.19 +/- 7.05 uM MDA versus 14.59 +/- 0.82 uM MDA, p < 0.05). Expression of the anti oxidants Aldo-keto reductase family 1 member C1 (AKR1C1) and Glutamate cysteine ligase catalytic subunit (GCLC) were significantly lower in GLC (p = 0.02) compared to NLC control. MMP was lower in GLC (57.5 +/- 6.8%) compared to NLC (41.8 +/- 5.3%). [Ca(2+)](i) levels were found to be higher (p < 0.001) in GLC cells compared to NLC. Expression of the plasma membrane Ca(2+)/ATPase (PMCA) and the sodium-calcium (NCX) exchangers were lower, while intracellular sarco endoplasmic reticulum Ca(2+)/ATPase 3 (SERCA) expression was significantly higher in GLC compared to NLC. Subjection of NLC cells to oxidative stress (200 uM H(2)0(2)) reduced expression of Na(+)/Ca2(+) exchanger 1 (NCX 1), plasma membrane Ca2+ ATPase 1 (PMCA 1), and PMCA 4 as determined by RT-PCR. CONCLUSIONS: Our data finds evidence of oxidative stress, mitochondrial dysfunction and impaired calcium extrusion in GLC cells compared to NLC cells and suggests their importance in the pathological changes occurring at the ONH in glaucoma. Future therapies may target reducing oxidative stress and / or [Ca(2+)](i). PMID- 21617753 TI - Microphakia and congenital cataract formation in a novel Lim2(C51R) mutant mouse. AB - PURPOSE: Within a mutagenesis screen, we identified the new mouse mutant Aca47 with small lenses and reduced axial eye lengths. The aim of the actual study was the molecular and morphological characterization of the mouse mutant Aca47. METHODS: We analyzed the offspring of paternally N-ethyl-N-nitrosourea (ENU) treated C57BL/6J mice for eye-size parameters by non-invasive in vivo laser interference biometry. Linkage analysis of the eye size mutant Aca47 was performed using single nucleotide polymorphisms and microsatellite markers. The Aca47 mutation was identified by sequence analysis of positional candidate genes. A general polymorphism at the mutated site was excluded by restriction analysis. Eyes of the Aca47 mouse mutant were characterized by histology. Visual properties were examined in the virtual drum. RESULTS: We identified a new mutant characterized by a significantly smaller lens and reduced axial eye length without any changes for cornea thickness, anterior chamber depth or aqueous humor size. The smaller size of lens was more pronounced in the homozygous mutants, which further developed congenital cataracts in the lens nucleus. The mutation was mapped to chromosome 7 between the markers D7Mit247 and D7Mit81. Using a positional candidate approach, the lens intrinsic integral membrane protein MP19 encoding gene Lim2 was sequenced; a T -> C exchange at cDNA position 151 leads to a cysteine-to-arginine substitution at position 51 of the Lim2 protein. Eye histology of adult heterozygous mutants did not show alterations on the cellular level. However, homozygous lenses revealed irregularly arranged lens fiber layers in the cortex. Virtual vision tests indicated that visual properties are not affected by reduced eye size of heterozygous individuals. CONCLUSIONS: These findings demonstrate a novel missense mutation in the Lim2 gene that affects lens development in a semidominant manner. Since homozygous mutants develop congenital lens opacities, this line can be used as a model for inherited cataract formation in humans. PMID- 21617754 TI - FKBP51 protects 661w cell culture from staurosporine-induced apoptosis. AB - PURPOSE: Neurodegenerative diseases and neurotraumas typically result in apoptosis of specific neurons leading to the pathology observed during the disease state. Existing treatments target the symptoms instead of preventing the death of these neurons. Although neuroprotective drugs should be useful as a treatment to prevent further loss of neurons, efficacious molecules are lacking. FK506 (tacrolimus), a widely used immunosuppressant drug, has significant neuroprotective and neuroregenerative properties throughout the central nervous system, including the eye. FK506 achieves these properties through interaction with FK506 binding proteins (FKBP), including FK506 binding protein 51 (FKBP51). In this study, we examine the effects of FKBP51 as a neuroprotective agent on a neuronal cell line. METHODS: We cultured 661w cell cultures with or without FK506, or stably transfected them with an FKBP51 expression vector. These cells were then exposed to the apoptosis-inducing agent staurosporine. Cell viability was determined using a calcein AM/propidium iodide assay. Protein levels and activation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF kappaB) were determined by western immunoblot analysis. RESULTS: FKBP51 overexpression significantly protected 661w cell cultures from staurosporine induced apoptosis. FKBP51 overexpression also significantly increased NF-kappaB p65 protein levels and activated NF-kappaB p65. FK506 treatment significantly protected 661w neuronal cultures from staurosporine-induced apoptosis. FK506 increased FKBP51, NF-kappaB p65, and levels of activated NF-kappaB p65 protein. CONCLUSIONS: These results suggest that FKBP51 protects 661w cell cultures from apoptosis induced by staurosporine. Additionally, FK506 protected 661w cell cultures from apoptosis and displayed a mechanism similar to that of FKBP51 overexpression. Both FK506 and FKBP51 appear to act through activation of NF kappaB p65 protein, suggesting a common pathway for neuroprotection. These findings implicate FKBP51 as a protein important to neuronal cell culture survival. FKBP51 may be a potential therapeutic drug target for preventing the neurodegeneration and neurotrauma that occur during neurodegenerative diseases. PMID- 21617755 TI - Effect of PITX2 knockdown on transcriptome of primary human trabecular meshwork cell cultures. AB - PURPOSE: To identify genes whose expressions in primary human trabecular meshwork (TM) cell cultures are affected by the transcription factor pituitary homeobox 2 (PITX2) and to identify genes that may have roles in glaucoma. Known glaucoma causing genes account for disease in a small fraction of patients, and we aimed at identification of other genes that may have subtle and accumulative effects not easily identifiable by a genetic approach. METHODS: Expression profiles derived using microarrays were compared between TM control cells and cells treated with PITX2 siRNAs using three protocols so as to minimize false positive and negative results. The first protocol was based on the commonly used B statistic. The second and third protocols were based on fold change in expression. The second protocol used a threshold of at least 2 fold change in expression, whereas the third protocol used ranking in fold change without setting a threshold. The likelihood of a selected gene being a true positive was considered to correlate with the number of protocols by which it was selected. By considering all genes that were selected by at least one protocol, the likelihood of false negatives was expected to decrease. Effects on a subset of selected genes were verified by real time PCR, western blots, and immunocytochemistry. Effects on ALDH1A1, were further pursued because its protein product, aldehyde dehydrogenase 1 family, member A1, has roles in oxidative stress and because oxidative stress is known to be relevant to the etiology of glaucoma. RESULTS: The expression level of 41 genes was assessed by to be possibly affected by PITX2 knockdown. Twenty one genes were down-regulated and twenty were upregulated. The expression of five genes was assessed to be altered by all three analysis protocols. The five genes were DIRAS3 (DIRAS family, GTP-binding RAS-like 3), CXCL6 (chemokine (C-X-C motif) ligand 6), SAMD5 (sterile alpha motif domain containing 5), CBFB (core-binding factor, beta subunit), and MEIS2 (meis homeobox 2). Real time PCR experiments verified results on a subset of genes tested. Notably, the results were also confirmed in two independent TMs. Effects on CXCL6 and ALDH1A1 were also confirmed by western blots, and effects on ALDH1A1 were further shown by immunocytochemistry. Data consistent with PITX2 involvement in ALDH1A1 mediated response to oxidative stress were presented. CONCLUSIONS: Bioinformatics tools revealed that the genes identified affect functions and pathways relevant to glaucoma. Involvement of PITX2 in expression of some of the genes and in some of the pathways is being reported here for the first time. As many of the genes identified have not been studied vis-a-vis glaucoma, we feel they introduce new candidates for understanding this devastating disease. PMID- 21617756 TI - Is the variant c.422+90G -> A in intron 4 of indoleamine 2, 3 -dioxygenase (IDO) gene related to age related cataracts? AB - PURPOSE: To screen for sequence variations in the IDO gene that encodes indoleamine 2, 3- dioxygenase (IDO), the first rate limiting enzyme involved in the tryptophan catabolism which results in the production of UV filters playing a role in the maintenance of lens transparency. METHODS: We conducted a case control study to screen for sequence changes in the IDO gene and associated demographic risk factors in patients with nuclear (NC-110), cortical (CC-110) and Posterior sub capsular (PSC-111) cataracts in comparison to normal controls (210) from Hyderabad, India. RESULTS: Among the cataract types studied high risk was observed for CC and PSC types in female patients, individuals with low body mass index and smoking habit. Cataract development had early onset more frequently in cases of PSC followed by CC and NC. Screening by single strand conformation polymorphism (SSCP) revealed mobility shifts in 6 of the 331 patient (3 with NC and 3 with PSC) samples which upon sequencing confirmed the presence of G -> A transition (c.422+90G -> A; rs4613984) in the intron downstream to exon 4 of IDO which was further tested by RFLP analysis using the HhaI restriction enzyme. Of the 6 patients, one with nuclear cataract showed homozygosity and the remaining five showed heterozygosity for the substitution. None of the control samples showed this variation. CONCLUSIONS: It is possible that the substitution c.422+90G -> A; rs4613984 in an intron downstream to exon 4 of IDO may be related with cataract formation among the aged. PMID- 21617757 TI - Transcriptional factors associated with epithelial-mesenchymal transition in choroidal neovascularization. AB - PURPOSE: To investigate the transcriptional factors associated with epithelial mesenchymal transition (EMT) in choroidal neovascularization (CNV) secondary to age-related macular degeneration (AMD). METHODS: Paraffin sections of CNV obtained from patients with AMD (n = 12) were stained for transcriptional factors related to EMT, i.e., Snail, Slug, SIP1, and Twist. As a control, postmortem sections of ocular normal tissue were used. Furthermore, using a human retinal pigment epithelial (RPE) cell line (ARPE-19), reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence microscopy were performed to explore the cellular localization and expression levels of EMT-associated transcriptional factors upon cytokine stimulation. RESULTS: Of 12 specimens, 11 CNV tissues (91.6%) showed staining for Snail localized in cellular nuclei, particularly in those of RPE cells. Snail was strongly co-localized with alpha smooth muscle antigen (SMA) in RPE cells. In contrast, postmortem human retina showed no Snail staining in RPE cells. Other transcriptional factors, Slug, Twist and SIP1 were not detected in CNV or normal human retina. In ARPE-19 cells, RT PCR and immunofluorescence microscopy showed that Snail mRNA was upregulated by transforming growth factor (TGF)-beta and VEGF stimulation. Furthermore, TGF-beta induced relocalization of Snail to the nucleus in RPE cells. CONCLUSIONS: The current data indicate that Snail is a major transcriptional factor for EMT changes of RPE cells in human CNV. PMID- 21617758 TI - Introduction to a new book about health, lifestyle, and health promotion. PMID- 21617759 TI - Improvements in the Formation of Boron-Doped Diamond Coatings on Platinum Wires Using the Novel Nucleation Process (NNP). AB - In order to increase the initial nucleation density for the growth of boron-doped diamond on platinum wires, we employed the novel nucleation process (NNP) originally developed by Rotter et al. and discussed by others [1-3]. This pretreatment method involves (i) the initial formation of a thin carbon layer over the substrate followed by (ii) ultrasonic seeding of this "soft" carbon layer with nanoscale particles of diamond. This two-step pretreatment is followed by the deposition of boron-doped diamond by microwave plasma-assisted CVD. Both the diamond seed particles and sites on the carbon layer itself function as the initial nucleation zones for diamond growth from an H(2)-rich source gas mixture. We report herein on the characterization of the pre-growth carbon layer formed on Pt as well as boron-doped films grown for 2, 4 and 6 h post NNP pretreatment. Results from scanning electron microscopy, Raman spectroscopy and electrochemical studies are reported. The NNP method increases the initial nucleation density on Pt and leads to the formation of a continuous diamond film in a shorter deposition time than is typical for wires pretreated by conventional ultrasonic seeding. The results indicate that the pregrowth layer itself consists of nanoscopic domains of diamond and functions well to enhance the initial nucleation of diamond without any diamond powder seeding. PMID- 21617760 TI - Stationary digital breast tomosynthesis with distributed field emission X-ray tube. AB - Tomosynthesis requires projection images from different viewing angles. Using a distributed x-ray source this can be achieved without mechanical motion of the source with the potential for faster image acquisition speed. A distributed x-ray tube has been designed and manufactured specifically for breast tomosynthesis. The x-ray tube consists of 31 field emission x-ray sources with an angular range of 30 degrees . The total dose is up to 100mAs with an energy range between 27 and 45 kVp. We discuss the source geometry and results from the characterization of the first prototype. The x-ray tube uses field emission cathodes based on carbon nanotubes (CNT) as electron source. Prior to the manufacturing of the sealed x-ray tube extensive testing on the field emission cathodes has been performed to verify the requirements for commercial tomosynthesis systems in terms of emission current, focal spot size and tube lifetime. PMID- 21617761 TI - Expedient synthesis of alpha,alpha-dimethyl-beta-hydroxy carbonyl scaffolds via Evans' aldol reaction with a tertiary enolate. AB - An efficient synthetic methodology for 3-hydroxy-2,2-dimethyloctynoic acid (DHOYA) and several variants, which are increasingly common fragments encountered in bioactive marine cyanobacterial metabolites, was developed. These fragments were obtained in three steps via a tertiary aldol reaction utilizing an Evans' chiral auxiliary to afford the desired stereochemistry at the beta-hydroxy carbon. Thus far, this methodology has been successfully applied in determination of the absolute stereochemistry of eight cyanobacterial natural products, including the VGSC activator palymramide A. PMID- 21617762 TI - Relationships Between Physical and Non-Physical Forms of Intimate Partner Violence and Depression among Urban Minority Adolescent Females. AB - BACKGROUND: Little is known about intimate partner violence (IPV) and depression among low income, urban African American and Hispanic adolescent females. METHOD: Interviews with 102 urban African American and Hispanic adolescent females examined physical abuse, emotional/verbal abuse, and threats, and their unique and combined associations with depression. RESULTS: One-quarter of the sample experienced all three types of abuse. Non-physical forms of IPV were significantly associated with depression. CONCLUSIONS: Some urban adolescent females from lower income households experience high rates of IPV. Physical and non-physical forms of IPV are important in understanding and responding to depression in this population. PMID- 21617764 TI - Nonionic Block Copolypeptide Micelles Containing a Hydrophobic racemic-Leucine Core. AB - Block copolymer micelles have been used extensively as carriers for therapeutic drugs and diagnostic molecules. Here, we report the synthesis of nonionic, block copolypeptides, K(P) (x)(rac-L)(y), which have a "reversed" rod-coil structure composed of a hydrophilic, rod-like, alpha-helical segment attached to a disordered, racemic hydrophobic segment. The self assembly of these block copolypeptides in water was studied, and their compositions were optimized to identify a sample, K(P) (100)(rac-L)(10), which is able to form well defined micelles that are very stable against dilution, high temperatures, and various media. Micelle structure was determined using a combination of electron microscopy and dynamic light scattering measurements. The potential of these micelles as drug delivery carriers was evaluated by encapsulation of the anticancer drug camptothecin. The drug containing micelles were found to be stable. PMID- 21617763 TI - Synthesis and Structure-Activity Relationships of Benzothienothiazepinone Inhibitors of Protein Kinase D. AB - Protein kinase D (PKD) is a member of a novel family of serine/threonine kinases that regulate fundamental cellular processes. PKD is implicated in the pathogenesis of several diseases, including cancer. Progress in understanding the biological functions and therapeutic potential of PKD has been hampered by the lack of specific inhibitors. The benzoxoloazepinolone CID755673 was recently identified as the first potent and selective PKD inhibitor. The study of structure-activity relationships (SAR) of this lead structure led to further improvements in PKD1 potency. We describe herein the synthesis and biological evaluation of novel benzothienothiazepinone analogs. We achieved a ten-fold increase in the in vitro PKD1 inhibitory potency for the second generation lead kb-NB142-70 and accomplished a transition to an almost equally potent novel pyrimidine scaffold, while maintaining excellent target selectivity. These promising results will guide the design of pharmacological tools to dissect PKD function and pave the way for the development of potential anti-cancer agents. PMID- 21617765 TI - Exploring the neonatal salivary transcriptome: technical optimization and clinical applications. PMID- 21617766 TI - Propensity score modelling in observational studies using dimension reduction methods. AB - Conditional independence assumptions are very important in causal inference modelling as well as in dimension reduction methodologies. These are two very strikingly different statistical literatures, and we study links between the two in this article. The concept of covariate sufficiency plays an important role, and we provide theoretical justification when dimension reduction and partial least squares methods will allow for valid causal inference to be performed. The methods are illustrated with application to a medical study and to simulated data. PMID- 21617767 TI - Abronione, a rotenoid from the desert annual Abronia villosa. AB - A high-throughput phytochemical investigation of Abronia villosa afforded a new rotenoid designated abronione (1) along with the known compounds boeravinone C and lupeol. The structure of 1 was determined using NMR, MS, and optical analysis with < 400 ug of material. Compound 1 displayed moderate cytotoxicity against NCI H460 and HL-60 human cancer cell lines with IC(50) values of 14 and 36 uM, respectively. PMID- 21617768 TI - Competition supports integrated care (?). PMID- 21617770 TI - New substituted isocoumarins and Dihydroisocoumarins and their cytotoxic activities. AB - New isocoumarins were prepared in an efficient way from 2-iodobenzoic acid derivatives and hept-1-yne in a Sonogashira reaction, followed by spontaneous cyclization. Catalytic hydrogenation gave the corresponding dihydroisocoumarins. A 4-chloroisocoumarin was prepared on an alternative pathway. Some of the new compounds showed moderate cytotoxic activities against a human leukemia cell line (HL 60). PMID- 21617769 TI - Phytoestrogen biological actions on Mammalian reproductive system and cancer growth. AB - Phytoestrogens are a family of diverse polyphenolic compounds derived from nature plant that structurally or functionally mimic circulating estrogen in the mammalian reproductive system. They induce estrogenic and anti-estrogenic effects in the brain-pituitary-gonad axis (a principal endocrine system involving in reproductive regulation) and peripheral reproductive organs. The dichotomy of phytoestrogen-mediated actions elucidates that they play the biological activities via complex mechanisms and belong to various chemical classes. In comparison with their unobvious physiological functions in normal reproductive tissues, there are increasing investigations showing that phytoestrogen induces significant inhibitory effects on the growth of breast and ovarian cancers through different signaling pathways. This review summarized the results of the previous studies regarding principal signaling transductions for mediating the growth of the ovarian and breast cancers. Phytoestrogen potentially modulates the signaling molecules via: (1) blocking the nuclear and membrane estrogen receptors (ER), (2) interfering with the growth factor receptor, (3) inhibiting the G protein-coupled receptor in ER-deficient cells, (4) activating apoptosis and nullifying anti-apoptotic signals. PMID- 21617771 TI - Chemometric QSAR modeling and in silico design of antioxidant NO donor phenols. AB - An acceleration of free radical formation within human system exacerbates the incidence of several life-threatening diseases. The systemic antioxidants often fall short for neutralizing the free radicals thereby demanding external antioxidant supplementation. Therein arises the need for development of new antioxidants with improved potency. In order to search for efficient antioxidant molecules, the present work deals with quantitative structure-activity relationship (QSAR) studies of a series of antioxidants belonging to the class of phenolic derivatives bearing NO donor groups. In this study, several QSAR models with appreciable statistical significance have been reported. Models were built using various chemometric tools and validated both internally and externally. These models chiefly infer that presence of substituted aromatic carbons, long chain branched substituents, an oxadiazole-N-oxide ring with an electronegative atom containing group substituted at the 5 position and high degree of methyl substitutions of the parent moiety are conducive to the antioxidant activity profile of these molecules. The novelty of this work is not only that the structural attributes of NO donor phenolic compounds required for potent antioxidant activity have been explored in this study, but new compounds with possible antioxidant activity have also been designed and their antioxidant activity has been predicted in silico. PMID- 21617772 TI - The gramine route to pyrido[4,3-b]indol-3-ones - identification of a new cytotoxic lead. AB - A novel approach to 3-oxo-gamma-carbolines was worked out starting from methyl indol-2-ylacetate via a gramine derivative. After quaternization, ammonia and 4 methoxybenzylamine could be inserted giving appropriate 3-oxo-gamma-carbolines. Condensation with 2-chlorobenzaldehyde under microwave irradiation gave a 4-(2 chlorobenzyl)-3-oxo-gamma-carboline. N-methylation lead to a product with very promising antifungal and cytotoxic activities. PMID- 21617773 TI - Rapid simultaneous determination of telmisartan, amlodipine besylate and hydrochlorothiazide in a combined poly pill dosage form by stability-indicating ultra performance liquid chromatography. AB - A simple, precise and rapid stability-indicating ultra-performance liquid chromatography (UPLC) method is developed for the simultaneous quantitative determination of Telmisartan, Amlodipine besylate and Hydrochlorothiazide from their innovative poly pill combination drug product in the presence of degradation products. It involves a 100 mm x 2.1 mm, 1.7 MUm C-18 column. The separation is achieved on a simple gradient method. The mobile phase A contains a mixture of sodium perchlorate buffer pH 3.2 (0.053M): acetonitrile in the ratio 90:10, v/v, and mobile B contains a mixture of sodium perchlorate buffer pH 3.2 (0.053M): acetonitrile in the ratio 20:80, v/v. The flow rate is 0.6 mL min(-1) and the column temperature is maintained at 55 degrees C.The gradient program (T/%B) is set as 0/5, 1.2/5, 1.6/40, 4/40, 4.1/5 and 4.5/5. The detector wavelength is 271 nm for Hydrochlorothiazide and Telmisartan and 237 nm for Amlodipine. The retention times of Telmisartan, Amlodipine, and Hydrochlorothiazide are 3.6 minutes, 3.2 minutes and 0.9 minutes; respectively. The total runtime for the separation of the three active compounds and their degradation products is 4.5 minutes. The described method is validated with respect to system suitability, specificity, linearity, precision and accuracy. The precision of the assay method is evaluated by carrying out six independent assays of T, A and H (0.032 mg mL(-1) of T, 0.004 mg mL(-1) of A, 0.01 mg mL(-1) of H). The accuracy of the method is evaluated in triplicate at three concentration levels, i.e. 50%, 100% and 150% of target test concentration (0.64 mg mL(-1) of T, 0.08 mg mL(-1) of A, 0.2 mg mL(-1) of H). The described method is linear over the range, 16 to 48 MUg mL(-1) for T, 2 to 6 MUg mL(-1)A and 5 to 15 MUg mL(-1) for H. The method is fast and suitable for high-throughput analysis allowing the analysis of about 250 samples per working day. PMID- 21617774 TI - Optimization of forced degradation using experimental design and development of a stability-indicating liquid chromatographic assay method for rebamipide in bulk and tablet dosage form. AB - A novel stability-indicating RP-HPLC assay method was developed and validated for quantitative determination of rebamipide in bulk and tablet dosage form. Rebamipide (drug and drug product) solutions were exposed to acid and alkali hydrolysis, thermal stress, oxidation by hydrogen peroxide and photodegradation. Experimental design has been used during forced degradation to determine significant factors responsible for degradation and to obtain optimal degradation conditions. In addition, acid and alkali hydrolysis was performed using a microwave oven. The chromatographic method employed the HiQ sil C-18HS (250 * 4.6 mm; 5 MUm) column with mobile phase consisting of 0.02 M potassium phosphate (pH adjusted to 6.8) and methanol (40:60, v/v) and the detection was performed at 230 nm. The procedure was validated for specificity, linearity, accuracy, precision and robustness. There was no interference observed of excipients and degradation products in the determination of the active pharmaceutical ingredient. The method showed good accuracy and precision (intra and inter day) and the response was linear in a range from 0.5 to 5 MUg mL(-1). The method was found to be simple and fast with less trial and error experimentation by making use of experimental design. Also, it proved that microwave energy can be used to expedite hydrolysis of rebamipide. PMID- 21617775 TI - Development and validation of a stability indicating RP-UPLC method for determination of quetiapine in pharmaceutical dosage form. AB - The present work reports a stability indicating reversed phase ultra performance liquid chromatography (RP-UPLC) method for the quantitative determination of quetiapine in pharmaceutical dosage form. The chromatographic separation is performed on an Agilent Eclipse Plus C18, RRHD 1.8 MUm (50 mm x 2.1 mm) column using gradient elution. The optimized mobile phase consists of 0.1 % aqueous triethylamine (pH 7.2) as a solvent-A and 80:20 v/v mixture of acetonitrile and methanol as solvent-B. The eluted compounds are monitored at 252 nm wavelength using a UV detector. The developed method separates quetiapine from its five impurities/degradation products within a run time of 5 min. Stability indicating capability of the developed method is established by analyzing forced degradation samples in which the spectral purity of quetiapine is ascertained along with the separation of degradation products from analyte peak. The developed RP-UPLC method is validated as per International Conference on Harmonization (ICH) guidelines with respect to system suitability, specificity, precision, accuracy, linearity, robustness and filter compatibility. PMID- 21617776 TI - Stability indicating LC-method for estimation of paracetamol and lornoxicam in combined dosage form. AB - A simple, specific and stability indicating reversed phase high performance liquid chromatographic method was developed for the simultaneous determination of paracetamol and lornoxicam in tablet dosage form. A Brownlee C-18, 5 MUm column having 250*4.6 mm i.d. in isocratic mode, with mobile phase containing 0.05 M potassium dihydrogen phosphate:methanol (40:60, v/v) was used. The flow rate was 1.0 ml/min and effluents were monitored at 266 nm. The retention times of paracetamol and lornoxicam were 2.7 min and 5.1 min, respectively. The linearity for paracetamol and lornoxicam were in the range of 5-200 MUg/ml and 0.08-20 MUg/ml, respectively. Paracetamol and lornoxicam stock solutions were subjected to acid and alkali hydrolysis, chemical oxidation and dry heat degradation. The proposed method was validated and successfully applied to the estimation of paracetamol and lornoxicam in combined tablet dosage form. PMID- 21617777 TI - Formulation and bioequivalence of two valsartan tablets after a single oral administration. AB - The aim of this study is to assess the quality of Valzan((r)) tablet (160 mg, valsartan immediate release test formulation) by comparing its pharmacokinetic parameters with Diovan((r)) tablet (160 mg, valsartan reference formulation). Valzan((r)) tablets were prepared according to a dry granulation method (roll compaction). To assess the bioequivalence of Valzan((r)) tablets a randomized, two-way, crossover, bioequivalence study was performed in 24 healthy male volunteers. The selected volunteers were divided into two groups of 12 subjects. One group was treated with the reference formulation (Diovan((r))) and the other one with the generic Valzan((r)), with a cross-over after the drug washout period of 14 days. Blood samples were collected at fixed time intervals and valsartan concentrations were determined by a validated HPLC assay method. The pharmacokinetic parameters AUC(0-48), AUC(0-infinity), C(max), T(max), K(e) and T(1/2) were determined for both the tablets and were compared statistically to evaluate the bioequivalence between the two brands of valsartan, using the statistical model recommended by the FDA. The analysis of variance (ANOVA) did not show any significant difference between the two formulations and 90% confidence intervals (CI) fell within the acceptable range for bioequivalence. Based on this statistical evaluation it was concluded that the test tablets (Valzan((r))) is well formulated, since it exhibits pharmacokinetic profile comparable to the reference brand Diovan((r)). PMID- 21617778 TI - Antitrypanosomal and cytotoxic activities of 22-Hydroxyclerosterol, a new sterol from Allexis cauliflora (Violaceae). AB - In the search for new antiparasitic natural compounds from the medicinal plants from Cameroon, the new 22-hydroxyclerosterol, established as such on the basis of detailed chemical and spectroscopic analysis, was isolated from the hexane extract of the stem bark of Allexis cauliflora together with the known clerosterol. 22-Hydroxyclerosterol inhibited the growth of Trypanosoma brucei brucei cells with an ED(50) value of 1.56 MUM. The compound was also established as an uncompetitive inhibitor of the glycolytic enzyme PGI of T. brucei (Ki'= 3 +/- 1 MUM), an uncompetitive inhibitor of mammalian rabbit muscles' enzyme PyK (Ki'= 26 +/- 3 MUM) and a mixed inhibitor of PyK of Leishmania mexicana (Ki'= 65 +/- 10 MUM; Ki= 24 +/- 5 MUM). PMID- 21617779 TI - Preparation of herbal tea as infusion or by maceration at room temperature using mistletoe tea as an example. AB - Herbal tea can be prepared by infusion or maceration at room temperature resulting in different compositions of extractable constituents, which possibly influences the mode of action or safety profile. Knowledge on this topic is limited. The aim of this study was to investigate the substantial differences between infusion and maceration as recommended preparation methods for the preparation of herbal mistletoe tea, a traditional remedy against cardiovascular diseases. No active substances are known but analytical marker substances such as proteins, triterpenoids, phenylpropane derivatives and flavonoids can be quantified within the herb and the different herbal tea preparations. Whereas phenylpropane derivatives were completely extracted by infusion and maceration, neither method dissolved viscotoxins. 43% of mistletoe lectins were extracted by maceration, whereas by infusion they are inactivated by thermal degradation. By contrast, oleanolic acid and betulinic acid are present in higher concentrations in infusates compared with macerates, but even infusion extracted less than 2%. Infusion extracted 43% of flavonoid-like substances and maceration only 31%. In conclusion this study determines some differences between both extraction methods on the profile of solved substances. The relevance of it should be determined in studies dealing with the efficacy of herbal mistletoe tea. PMID- 21617780 TI - Anatomy of subterranean organs of medicinally used cardueae and related species and its value for discrimination. AB - Numerous species of the Asteraceae, the composites, are famous for their use in both traditional and conventional medicine. Reliable anatomical descriptions of these plants and of possible adulterations provide a basis for fast identification and cheap purity controls of respective medicinal drugs by means of light microscopy. Nevertheless, detailed comparative studies on root and rhizome anatomy of valuable as well as related inconsiderable composite plants are largely missing yet. The presented study aims to narrow this gap by performing anatomical analyses of roots and rhizomes of 16 species belonging to the tribe Cardueae, of formerly and currently used drugs as well as their near relatives as potential adulterations (Carlina acaulis L., Carlina vulgaris L., Arctium lappa L., Arctium tomentosum Mill., Carduus defloratus L., Carduus personata (L.) Jacq, Cirsium arvense (L.) Scop., Cirsium vulgare (Savi) Ten., Cirsium erisithales (Jacq.) Scop., Onopordum acanthium L., Silybum marianum (L.) Gaertn., Rhaponticum scariosum Lam., Centaurea jacea L., Centaurea scabiosa L., Centaurea cyanus L., Cnicus benedictus L.). A detailed verbal and graphical survey of the analysed anatomical features is provided. Several characters were finally extracted which allow for discrimination of the examined species and may be effectively used for drug quality controls. PMID- 21617781 TI - Phenolics from Rhagadiolus stellatus (Asteraceae, Cichorieae). AB - Rhagadiolus stellatus Gaertn., a Mediterranean member of the Cichorieae tribe of the Asteraceae family used as a food plant, was analyzed for its spectrum of phenolic compounds. Kaempferol 3-O-beta-glucoside 1, kaempferol 3-O-beta rutinoside (nicotiflorin) 2, quercetin 3-O-beta-glucoside 3, and luteolin 4 were isolated from the n-butanol layer of a methanolic extract of whole plants of Rh. stellatus of Spanish origin by repeated Sephadex LH-20 column chromatography. Structures were determined based on NMR and MS data as well as by comparison with literature data. Additionally, chlorogenic acid 5 and 3,5-dicaffeoylquinic acid 6 were detected by HPLC/DAD and HPLC/MS. Chemosystematic implications of the presented findings are discussed in comparison with other members of the Cichorieae tribe. PMID- 21617782 TI - Optimization, characterisation and pharmacokinetic studies of mucoadhesive oral multiple unit systems of ornidazole. AB - The objective of the present study was to investigate the applicability of matrix type mucoadhesive oral multiple unit systems (MUS) for sustaining the release of ornidazole in the gastrointestinal tract (GIT). The MUS were prepared by ionotropic gelation method using chitosan and hydroxypropyl methyl cellulose K4M (HPMC K4M) according to 3(2) factorial designs and were evaluated in vitro and in vivo. The particle size length ranged from 0.78 to 1.30 mm and breadth from 0.76 to 1.30 mm, respectively. The entrapment efficiency was in range of 80 to 96%. The rapid wash-off test was observed faster at intestinal pH 6.8 as compared to acidic pH 1.2. The fluoroscopic study revealed the retention of MUS in GIT for more than 5 hours. The pharmacokinetic parameters C(max), T(max), mean residence time (MRT) and area under curve (AUC) of developed MUS were found to be improved significantly (p<0.05) when compared with marketed immediate release tablets each containing 500 mg of drug. This study demonstrates that the MUS could be a good alternative to immediate release tablets to deliver ornidazole and expected to be less irritant to gastric and intestinal mucosa. PMID- 21617784 TI - The use of liquid-based cytology in cervical cancer screening. PMID- 21617783 TI - Preparation of mucoadhesive oral patches containing tetracycline hydrochloride and carvacrol for treatment of local mouth bacterial infections and candidiasis. AB - The specific aim of this work was to prepare mucoadhesive patches containing tetracycline hydrochloride and carvacrol in an attempt to develop a novel oral drug delivery system for the treatment of mouth infections. The bilayered patches were prepared using ethyl cellulose as a backing layer and carbopol 934 as a matrix mucoadhesive layer. Patches were prepared with different loading amounts of tetracycline hydrochloride and carvacrol. The antimicrobial activity was assessed for the prepared patches using the disc-diffusion method against the yeast Candida albicans and five bacterial strains, including Pseudomonas aeruginosa, Escherichia coli, Bacillus cereus, Staphylococcus aureus, and Bacillus bronchispti. In this work, we highlighted the possibility of occurrence of a synergistic action between carvacrol and tetracycline. The best formulation was selected based on microbiological tests, drug release, ex-vivo mucoadhesive performance, and swelling index. Physical characteristics of the selected formulations were determined. These included pH, patch thickness, weight uniformity, content uniformity, folding endurance, and patch stability. PMID- 21617785 TI - The impact of liquid-based cytology in decreasing the incidence of cervical cancer. AB - Major advances in screening have lowered the death rate from cervical cancer in the United States. One of the first major advances in cervical cancer screening was the Papanicolaou (Pap) test. The second major advance was liquid-based cytology (LBC). This review presents a wide range of data, discusses the strengths and weaknesses of the available information regarding Pap technologies, and reviews the meta-analyses, which have examined the differences in clinical performance. The review concludes with information on new and future developments to further decrease cervical cancer deaths. PMID- 21617786 TI - [Extravasation of cytostatic drugs]. AB - Paravasation of cytostatic drugs during peripheral intravenous administration is a well known complication. In the United States of America it occurs in seven percent of cases with different severity and consequences. Although methods to completely avoid this complication are still unavailable, we are able to decrease the risks by identifying the patient- and procedure-related factors. The educated patient is a good indicator of paravasation in case he or she can cooperate and call the nurse. When the patient is unable to cooperate, the risks of extravasation is higher and closer nursing surveillance is indicated. The extent of injury depends mainly on the chemical structure of the extravasant substance (vesicant, irritant or non-vesicant) which may be modified by other factors. There is no strong evidence-based guidance for the management of complication. Abrupt cessation of the infusion and drawing back on the inserted venous catheter as well as elevating and resting the affected limb are necessary measures. In the available literature cooling or warming of the affected area is controversial. Similarly there are still open questions regarding the value of using antidotes as dexrazoxane, dimethylsulfoxide, thiosulfate and hyaluronidase (which is not registered as medicament in Hungary). In the event of extravasation early multidisciplinary dermatological and surgical assessment is essential for definitive diagnosis and setting the optimal management. PMID- 21617787 TI - [Introduction of a clinical protocol for extravasation at the National Institute of Oncology, Budapest, Hungary]. AB - Extravasation of cytostatics occurs when an infusion containing a cytotoxic drug leaks into the surrounding perivascular and subcutaneous tissues. Incidence of cytostatic extravasation is found to be 0.1-6% according to the literature. Depending on the severity of complications, pain, loss of function in the extremities, or in extreme cases tissue necrosis necessitating an amputation may develop, drawing consequences like delay or interruption of the chemotherapy. Extent of complications is greatly influenced by the type of medication administered, general condition of the patient, and professional preparedness of staff providing the oncological health service. The protocol recently implemented in the National Institute of Oncology is a short, compact guidance for physicians and nurses providing oncological care, so by quick and adequate management of extravasation cases, severe complications could be prevented. More complex practical guidelines including algorithms could be created as a result of a wider collaboration, with the help of which oncological health professionals could easily cope with this rare problem. The authors describe in their review the implementation of the use of dry warm and cold packs, dymethylsulfoxide and hyaluronidase and their function within the algorithm of extravasation treatment. PMID- 21617788 TI - [Possibilities of hypnosis and hypnosuggestive methods in oncology]. AB - Fear of death, pain, or the recurrence of the illness of tumor patients can narrow their attention to a point where a spontaneous altered state of consciousness occurs. In these cases hypnosis either in formal psychotherapy or embedded into the everyday communication with the physician can effectively complement other already known medical and psychological techniques. Although numerous studies have reported the beneficial physical and mental changes induced by hypnosis, for a long time there were not enough research to affect evidence based medicine. New studies meeting the most rigorous methodological standards, new reviews and the characteristics of hypnosis shown by neuroimaging techniques support the acceptance of this method. Hypnosis is used and studied with adult and child tumor patients alike mostly in the areas of anxiety, pain, nausea, vomiting, quality of life, mood amelioration, immune system and hot flushes. Most of the assays describe hypnosis as an empirically validated treatment technique that in most cases surpass attention diversion, coping trainings, cognitive behavior and relaxation techniques and other regular treatments. In this paper we review these observations. PMID- 21617789 TI - [Aromatase inhibitors and arthralgia]. AB - In several large adjuvant clinical trials it has been demonstrated that substitution (eventually addition) of aromatase inhibitors (AIs) provides an improved outcome of endocrine-sensitive breast cancer over tamoxifen alone. Nevertheless, arthralgia induced by the AIs is one of the most frequent side effects in hormonal therapy. It is characterized by tenosynovial changes and is more frequent in patients in clinical practice than previously appreciated in adjuvant clinical trials. AI-related arthralgia may be related to estrogen deprivation, but estrogen replacement is not an option for these women. Therefore standard painkillers, NSAIDs (COX2 inhibitors), week opioids and other interventions (vitamin D, calcium, bisphosphonates, exercise, acupuncture, complementary and alternative approaches, eventually switch to another endocrine drug) are used for managing this treatment-related side effect, and improve adherence and quality of life among breast cancer survivors. PMID- 21617790 TI - [Challenges in oncologic plastic surgery of the breast]. AB - Breast screening programs along with advances in diagnostic methods and oncologic treatment have resulted in full recovery for a decisive number of patients diagnosed with early-stage breast cancer. The results of the ultra-radical-, followed by the breast conserving era pose new opportunities and challenges for the oncologic breast surgeon. The focus of oncoplastic surgery is not only on the tumor, but also on the female patient, allowing for individualized immediate breast reconstruction with acceptable esthetic result following radical tumor exstirpation. Modern procedures differ both in concept and technique from that of traditional breast surgery. This paper provides a comprehensive and detailed overview of reconstructive and oncoplastic breast surgery. PMID- 21617791 TI - [Study of motogenic signal in human melanoma cells]. AB - The components of the extracellular matrix (ECM) are more than just adhesion sites for migrating tumor cells: following enzymatic degradation of the ECM, the release of sequestrated growth factors increases, thus they become available for tumor cells. In a number of cancers dysfunction of epidermal growth factor receptor (EGFR) or hepatocyte growth factor receptor (c-Met) contribute to the malignant transformation that directly regulates cell proliferation, survival and motility. Furthermore, intracellular calcium level plays an important role in the regulation of the tyrosine kinase pathway. In our preclinical experiments, by administering heparin-derived oligosaccharides we influenced the interaction between human melanoma cells and ECM. In vitro cell migration was inhibited by heparin fragments. Moreover, two of the effective oligosaccharides reduced the number of lung colonies formed in SCID mice. In human melanoma cells an important element of Ca2+ homeostasis, the purinergic Ca2+ channel P2X7 proved to be an anti-apoptotic protein. EGFR and c-Met showed constitutive activity in human melanoma cells, and their inhibition in vitro caused decreased proliferation, migration and elevated apoptosis. Administration of a selective c-Met-TKI significantly decreased primary tumor growth in vivo as well as the capacity for liver colony formation in SCID mice. Selective EGFR-TKI had less inhibitory effect on metastasis formation, and had no effect on the primary tumor. Our results suggest the necessity of a rational dual-specific drug design for the purpose in the therapy of malignant melanoma. PMID- 21617792 TI - New photocatalyst BiOCl/BiOI composites with highly enhanced visible light photocatalytic performances. AB - BiOCl/BiOI composites with a visible light response were prepared by a simple hydrothermal method. Even though both single BiOCl and BiOI show low photocatalytic activity, BiOCl/BiOI composites provide enhanced efficiency in decomposing organic compounds including Methyl Orange (MO) and Rhodamine B (RhB). Furthermore, the 20%BiOCl/BiOI composite shows the highest efficiency for decomposing MO, while the highest performance is observed for the degradation of RhB over 70%BiOCl/BiOI composite. A possible photocatalytic mechanism has been proposed based on the relative experiments and the band positions of BiOCl and BiOI. PMID- 21617793 TI - In vitro 3D collective sprouting angiogenesis under orchestrated ANG-1 and VEGF gradients. AB - Sprouting angiogenesis requires a coordinated guidance from a variety of angiogenic factors. Here, we have developed a unique hydrogel incorporating microfluidic platform which mimics the physiological microenvironment in 3D under a precisely orchestrated gradient of soluble angiogenic factors, VEGF and ANG-1. The system enables the quantified investigation in chemotactic response of endothelial cells during the collective angiogenic sprouting process. While the presence of a VEGF gradient alone was sufficient in inducing a greater number of tip cells, addition of ANG-1 to the VEGF gradient enhanced the number of tip cells that are attached to collectively migrated stalk cells. The chemotactic response of tip cells attracted by the VEGF gradient and the stabilizing role of ANG-1 were morphologically investigated, elucidating the 3D co-operative migration of tip and stalk cells as well as their structures. We found that ANG-1 enhanced the connection of the stalk cells with the tip cells, and then the direct connection regulated the morphogenesis and/or life cycle of stalk cells. PMID- 21617794 TI - Photolytic processing of secondary organic aerosols dissolved in cloud droplets. AB - The effect of UV irradiation on the molecular composition of aqueous extracts of secondary organic aerosol (SOA) was investigated. SOA was prepared by the dark reaction of ozone and d-limonene at 0.05-1 ppm precursor concentrations and collected with a particle-into-liquid sampler (PILS). The PILS extracts were photolyzed by 300-400 nm radiation for up to 24 h. Water-soluble SOA constituents were analyzed using high-resolution electrospray ionization mass spectrometry (HR ESI-MS) at different stages of photolysis for all SOA precursor concentrations. Exposure to UV radiation increased the average O/C ratio and decreased the average double bond equivalent (DBE) of the dissolved SOA compounds. Oligomeric compounds were significantly decreased by photolysis relative to the monomeric compounds. Direct pH measurements showed that acidic compounds increased in abundance upon photolysis. Methanol reactivity analysis revealed significant photodissociation of molecules containing carbonyl groups and the formation of carboxylic acids. Aldehydes, such as limononaldehyde, were almost completely removed. The removal of carbonyls was further confirmed by the UV/Vis absorption spectroscopy of the SOA extracts where the absorbance in the carbonyl n->pi* band decreased significantly upon photolysis. The effective quantum yield (the number of carbonyls destroyed per photon absorbed) was estimated as ~0.03. The total concentration of peroxides did not change significantly during photolysis as quantified with an iodometric test. Although organic peroxides were photolyzed, the likely end products of photolysis were smaller peroxides, including hydrogen peroxide, resulting in a no net change in the peroxide content. Photolysis of dry limonene SOA deposited on substrates was investigated in a separate set of experiments. The observed effects on the average O/C and DBE were similar to the aqueous photolysis, but the extent of chemical change was smaller in dry SOA. Our results suggest that biogenic SOA dissolved in cloud and fog droplets will undergo significant photolytic processing on a time scale of hours to days. This type of photolytic processing may account for the discrepancy between the higher values of O/C measured in the field experiments relative to the laboratory measurements on SOA in smog chambers. In addition, the direct photolysis of oligomeric compounds may be responsible for the scarcity of their observation in the field. PMID- 21617795 TI - On the interaction of ascorbic acid and the tetrachlorocuprate ion [CuCl4]2- in CuCl nanoplatelet formation from an ionic liquid precursor (ILP). AB - The formation of CuCl nanoplatelets from the ionic liquid precursor (ILP) butylpyridinium tetrachlorocuprate [C(4)Py](2)[CuCl(4)] using ascorbic acid as a reducing agent was investigated. In particular, electron paramagnetic resonance (EPR) spectroscopy was used to evaluate the interaction between ascorbic acid and the Cu(II) ion before reduction to Cu(I). EPR spectroscopy suggests that the [CuCl(4)](2-) ion in the neat IL is a distorted tetrahedron, consistent with DFT calculations. Addition of ascorbic acid leads to the removal of one chloride from the [CuCl(4)](2-) anion, as shown by DFT and the loss of symmetry by EPR. DFT furthermore suggests that the most stable adduct is formed when only one hydroxyl group of the ascorbic acid coordinates to the Cu(II) ion. PMID- 21617796 TI - The origin of dips for the graphene-based DNA sequencing device. AB - We recently proposed an ultrafast DNA sequencing method that electrically distinguishes different nucleobases on a graphene nanoribbon (GNR), utilizing pi pi interaction. Analyzing the molecular orbitals (MOs) and the features of dips in conductance for our GNR-based sequencing device, we prove that the Fano resonance is responsible for the characteristic dips of each nucleobase. PMID- 21617797 TI - Pneumatically tunable optofluidic 2 * 2 switch for reconfigurable optical circuit. AB - We presented a pneumatically tunable 2 * 2 optofluidic switch for on-chip light routing that was controlled by compressed air. The device was fabricated with an optically clear elastomer-polydimethylsiloxane (PDMS)-by soft-lithography. The optical switching is realized with a tunable air-gap mirror by which the light is deflected due to total internal reflection in the bypass state. When the device is subjected to high pressure, the air gap collapses and hence the light will be switched to the crossover state. The device had a switching speed of more than 5 Hz and an extinction ratio of 8 dB. This switch can be readily integrated with other microfluidic circuits. We demonstrated a simple reconfigurable optical waveguide circuit for dual-channel microfluidic spectroscopy measurement on a chip. PMID- 21617798 TI - Application of platinum nanoparticles as affinity probe and matrix for direct analysis of small biomolecules and microwave digested proteins using matrix assisted laser desorption/ionization mass spectrometry. AB - We report the use of platinum nanoparticles (PtNPs) for analysis of amino acids, peptides, proteins and microwave digested proteins (lysozyme and bovine serum albumin) without any tedious washing and separation procedures prior to matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). In the present study, PtNPs play three functions, such as matrix, affinity probe and acceleration of protein digestion by absorbing the microwave irradiation. Good signal intensity of the target molecules from the sample was obtained when laser energy, NPs concentration and incubation time were set to 35 MUJ, 25 nM and 30 min, respectively. In addition, higher numbers of peptide sequence were obtained for microwave digested lysozyme protein using PtNPs as compared to previously reported methods for analysis of digested protein in MALDI MS. Thus, the present method is a simple, rapid and one step preparation method for the analysis of amino acids, peptides, proteins and digested proteins in MALDI-TOF-MS without the need for any tedious purifications and washing procedures. PMID- 21617799 TI - Density functional study on the increment of carrier mobility in armchair graphene nanoribbons induced by Stone-Wales defects. AB - Armchair graphene nanoribbons and their derived structures containing Stone-Wales defects are investigated using a self-consistent field crystal orbital method based on density functional theory. The investigation indicates that both the nanoribbons and the defective structures are semiconductors. A low concentration of middle Stone-Wales defects generally increases the carrier mobility, calculated using deformation potential theory, while edge Stone-Wales defects decrease it. The largest increment of the carrier mobility is as high as 170%, which is explained by the lighter carrier effective mass with crystal orbital analysis. PMID- 21617800 TI - Boron environments in Pyrex(r) glass--a high resolution, Double-Rotation NMR and thermodynamic modelling study. AB - It is shown, using the important technological glass Pyrex(r) as an example, that 1D and 2D (11)B Double-Rotation (DOR) NMR experiments, in combination with thermodynamic modelling, are able to provide unique structural information about complex glasses. (11)B DOR NMR has been applied to Pyrex(r) glass in order to remove both dipolar and quadrupolar broadening of the NMR lines, leading to high resolution spectra that allow unambiguous, accurate peak fitting to be carried out, of particular importance in the case of the 3-coordinated [BO(3)] (B3) trigonal planar environments. The data obtained are of sufficient quality that they can be used to test the distributions of borate and borosilicate superstructural units predicted by the thermodynamics-based Model of Associated Solutions. The model predicts the dominant boron-containing chemical groupings in Pyrex(r) glass to be those associated with B(2)O(3) and sodium tetraborate (with smaller amounts of sodium triborate, sodium diborate, sodium pentaborate, danburite and reedmergnerite). Excellent agreement is found between model and experiment provided the (11)B peaks with isotropic chemical shifts of -1.4 ppm and 0.5 ppm are assigned to B4 species from borosilicate units ([B(OSi)(4)] and [B(OSi)(3)(OB)]) and borate superstructural units (mainly triborate rings with some pentaborate and diborate) respectively. The peaks with isotropic shifts of 14 ppm and 18.1 ppm are then assigned to B3 in borate superstructural units (mainly triborate and pentaborate along with connecting B3) and boroxol rings respectively. The assignments of the DOR NMR peaks, are supported by the presence of cross-peaks in (11)B spin-diffusion DOR NMR spectra which can be used to develop a structural model in which B(2)O(3)-like regions are linked, via borate and borosilicate superstructural units, to the majority silica network. Pyrex(r) is thus shown to have a heterogeneous structure, with distinct molecular groupings that are far removed from a random distribution of network polyhedra with only short-range order. PMID- 21617801 TI - Protein phosphorylation analysis in archival clinical cancer samples by shotgun and targeted proteomics approaches. AB - Protein phosphorylation affects most eukaryotic cellular processes and its deregulation is considered a hallmark of cancer and other diseases. Phosphoproteomics may enable monitoring of altered signaling pathways as a means of stratifying tumors and facilitating the discovery of new drugs. Unfortunately, the development of molecular tests for clinical use is constrained by the limited availability of fresh frozen, clinically annotated samples. Here we report phosphopeptide analysis in human archival formalin-fixed, paraffin-embedded (FFPE) cancer samples based on immobilized metal affinity chromatography followed by liquid chromatography coupled with tandem mass spectrometry and selected reaction monitoring techniques. Our results indicate the equivalence of detectable phosphorylation rates in archival FFPE and fresh frozen tissues. Moreover, we demonstrate the applicability of targeted assays for phosphopeptide analysis in clinical archival FFPE samples, using an experimental workflow suitable for processing and analyzing large sample series. This work paves the way for the application of shotgun and targeted phosphoproteomics approaches in clinically relevant studies using archival clinical samples. PMID- 21617802 TI - Solute-solvent friction kernels and solution properties of methyl oxazoline phenyl oxazoline (MeOx-PhOx) copolymers in binary ethanol-water mixtures. AB - Solvent mixtures often alter the solubility of polymeric substances. Statistical copolymers made from 2-methyl-2-oxazoline (MeOx) and 2-phenyl-2-oxazoline (PhOx) are known for their varying solubilities in pure ethanol, pure water and in binary mixtures of ethanol-water. Constrained Molecular Dynamics (MD) simulations have been carried out with an aim to explain the varying solubilities of the statistical MeOx-PhOx copolymers. The solute-solvent dynamic friction kernels calculated through constrained MD simulations corroborate the solubility pattern in these copolymers. The solvation characteristics have been analyzed in terms of the solute-solvent radial distribution functions (RDFs). The ethanol-soluble MeOx PhOx copolymers exhibit characteristic solute-composition dependence in the dynamic solute-solvent friction kernels, indicating the strength of the solute solvent correlations. The aggressive solvation by the ethanol molecules in the binary solvent mixtures has been brought out by the O(solute)-H(ethanol) RDFs which exhibit a characteristic dependence on the ethanol content in the solvent composition. The corresponding O(solute)-H(water) RDFs are devoid of any such composition dependence. For all the MeOx-PhOx copolymers, the O-site solvation is strongly dominated by the water molecules and the N-sites are solvated equally by both ethanol and water molecules. PMID- 21617803 TI - Urine analysis in microfluidic devices. AB - Microfluidics has attracted considerable attention since its early development in the 1980s and has experienced rapid growth in the past three decades due to advantages associated with miniaturization, integration and automation. Urine analysis is a common, fast and inexpensive clinical diagnostic tool in health care. In this article, we will be reviewing recent works starting from 2005 to the present for urine analysis using microfluidic devices or systems and to provide in-depth commentary about these techniques. Moreover, commercial strips that are often treated as chips and their readers for urine analysis will also be briefly discussed. We start with an introduction to the physiological significance of various components or measurement standards in urine analysis, followed by a brief introduction to enabling microfluidic technologies. Then, microfluidic devices or systems for sample pretreatments and for sensing urinary macromolecules, micromolecules, as well as multiplexed analysis are reviewed, in this sequence. Moreover, a microfluidic chip for urinary proteome profiling is also discussed, followed by a section discussing commercial products. Finally, the authors' perspectives on microfluidic-based urine analysis are provided. These advancements in microfluidic techniques for urine analysis may improve current routine clinical practices, particularly for point-of-care (POC) applications. PMID- 21617804 TI - Conformational polymorphs of 1,1,2,2-tetrachloroethane: pressure vs. temperature. AB - Directional Cl...Cl type I and II interactions govern the low-density aggregation of 1,1,2,2-tetrachloroethane molecules in synclinal conformation in the crystalline state at low temperature, whereas the dense molecular packing in high pressure is achieved for the antiperiplanar conformers and electrostatically less favored Cl...Cl contacts. The mechanism of transformation between loose and dense associations involves the collapse of Cl...Cl contacts and conformational conversion. PMID- 21617805 TI - Multiplexed color encoded silica nanospheres prepared by stepwise encapsulating quantum dot/SiO2 multilayers. AB - We present a flexible method for multiplexed colour encoded nanospheres (MENs) encapsulating layers of different colour quantum dots. Our method results in highly efficient photoluminescent nanospheres with monodisperse, photostable, and excellent luminescence properties. PMID- 21617806 TI - Plasmon-induced CD response of oligonucleotide-conjugated metal nanoparticles. AB - Non-chiral metal nanoparticles conjugated with chiral oligonucleotide molecules demonstrated a circular dichroism (CD) at the plasmonic wavelengths due to aggregation effects. PMID- 21617808 TI - A V-shaped cationic dye for nonlinear optical bioimaging. AB - A symmetric cationic molecule with D-pi-A(+)-pi-D architecture was synthesized with high two-photon absorption cross-section (sigma(2) ~ 1140 GM). Application as a marker in fluorescence microscopy of living cells revealed its presence inside the cell staining vesicular shape organelles in the cytoplasm. Fluorescence lifetime imaging microscopy shows that it is also able to penetrate within the nucleus. PMID- 21617807 TI - Fluorinated paramagnetic chelates as potential multi-chromic 19F tracer agents. AB - A class of potential multi-chromic (19)F imaging tracers is made by pairing metal ions with a fluorinated chelator. All fluorinated metal chelates emit a single (19)F signal. Paramagnetic metal ions shifted the (19)F signal frequency and made the (19)F relaxation rates insensitive toward local chemical environment. PMID- 21617809 TI - A rational route to SCM materials based on a 1-D cobalt selenocyanato coordination polymer. AB - Thermal annealing of a discrete complex with terminal SeCN anions and monodentate coligands enforces the formation of a 1D cobalt selenocyanato coordination polymer that shows slow relaxation of the magnetization. Therefore, this approach offers a rational route to 1D materials that might show single chain magnetic behaviour. PMID- 21617810 TI - Formation of stable neutral copper bis-dithiolene thin films by potentiostatic electrodeposition. AB - Thin films of neutral copper dithiolenes have been prepared by potentiostatic electrodeposition. This method allows the isolation of near infrared (NIR) active species, in a useable form, that are otherwise unobtainable by conventional chemical methods. PMID- 21617811 TI - Antibiotic activities of host defense peptides: more to it than lipid bilayer perturbation. AB - Defensins are small basic amphiphilic peptides (up to 5 kDa) that have been shown to be important effector molecules of the innate immune system of animals, plants and fungi. In addition to immune modulatory functions, they have potent direct antimicrobial activity against a broad spectrum of bacteria, fungi and/or viruses, which makes them promising lead compounds for the development of next generation antiinfectives. The mode of antibiotic action of defensins was long thought to result from electrostatic interaction between the positively charged defensins and negatively charged microbial membranes, followed by unspecific membrane permeabilization or pore-formation. Microbial membranes are more negatively charged than human membranes, which may explain to some extent the specificity of defensin action against microbes and associated low toxicity for the host. However, research during the past decade has demonstrated that defensin activities can be much more targeted and that microbe-specific lipid receptors are involved in the killing activity of various defensins. In this respect, human, fungal and invertebrate defensins have been shown to bind to and sequester the bacterial cell wall building block lipid II, thereby specifically inhibiting cell wall biosynthesis. Moreover, plant and insect defensins were found to interact with fungal sphingolipid receptors, resulting in fungal cell death. This review summarizes the current knowledge on the mode of action and structure of defensins from different kingdoms, with specific emphasis on their interaction with microbial lipid receptors. PMID- 21617812 TI - An efficient route to 1-aminoisoquinolines via AgOTf-catalyzed reaction of 2 alkynylbenzaldoxime with amine. AB - 2-Alkynylbenzaldoxime reacts with amine catalyzed by silver triflate under mild conditions, leading to 1-aminoisoquinolines in good yield. This reaction proceeds efficiently with good functional group tolerance. PMID- 21617813 TI - Exploiting Sm(II) and Sm(III) in SmI2-initiated reaction cascades: application in a tag removal-cyclisation approach to spirooxindole scaffolds. AB - A tag removal-cyclisation sequence is described that is initiated by reduction using a Sm(II) species and completed by a Sm(III) Lewis acid that is formed in an earlier stage. Therefore, the reaction cascade utilises both oxidation states of a samarium reagent in discrete steps and allows access to privileged, pyrrolidinyl-spirooxindole scaffolds and analogues inspired by the anti-cancer natural product spirotryprostatin A. PMID- 21617814 TI - Quantum clustering and network analysis of MD simulation trajectories to probe the conformational ensembles of protein-ligand interactions. AB - In this article, we present a novel application of a quantum clustering (QC) technique to objectively cluster the conformations, sampled by molecular dynamics simulations performed on different ligand bound structures of the protein. We further portray each conformational population in terms of dynamically stable network parameters which beautifully capture the ligand induced variations in the ensemble in atomistic detail. The conformational populations thus identified by the QC method and verified by network parameters are evaluated for different ligand bound states of the protein pyrrolysyl-tRNA synthetase (DhPylRS) from D. hafniense. The ligand/environment induced re-distribution of protein conformational ensembles forms the basis for understanding several important biological phenomena such as allostery and enzyme catalysis. The atomistic level characterization of each population in the conformational ensemble in terms of the re-orchestrated networks of amino acids is a challenging problem, especially when the changes are minimal at the backbone level. Here we demonstrate that the QC method is sensitive to such subtle changes and is able to cluster MD snapshots which are similar at the side-chain interaction level. Although we have applied these methods on simulation trajectories of a modest time scale (20 ns each), we emphasize that our methodology provides a general approach towards an objective clustering of large-scale MD simulation data and may be applied to probe multistate equilibria at higher time scales, and to problems related to protein folding for any protein or protein-protein/RNA/DNA complex of interest with a known structure. PMID- 21617815 TI - Simultaneous enhancement of chemical shift dispersion and diffusion resolution in mixture analysis by diffusion-ordered NMR spectroscopy. AB - Mixture analysis by high resolution diffusion-ordered NMR spectroscopy (HR-DOSY) requires differences in both chemical shift and diffusion coefficient; resolution can be greatly enhanced by exploiting the chemical specificity of lanthanide shift reagent binding to increase chemical shift and diffusion dispersion simultaneously. PMID- 21617816 TI - 4-Nitrophenyl sulfenyl chloride as a new precursor for the formation of aromatic SAMs on gold surfaces. AB - 4-Nitrophenyl sulfenyl chloride was used as a new precursor for the formation of densely packed aromatic SAMs on gold. The adsorption involves the reductive dissociation of the S-Cl bond. A well-ordered row structure corresponding to a ?3 * 4 phase with a molecular area of 27.8 A(2) is obtained. PMID- 21617817 TI - A near-infrared fluorescent probe for detecting copper(II) with high selectivity and sensitivity and its biological imaging applications. AB - The first near-infrared fluorescent probe was developed toward Cu(2+). Based on the photo-induced electron transfer (PET) mechanism, the probe exhibited weak fluorescence. Upon the addition of Cu(2+), it fluoresced strongly. The probe offered this unique capability, and was successfully applied to living cells, tissues and in vivo to visualize Cu(2+). PMID- 21617818 TI - Engineering of a bis-chelator motif into a protein alpha-helix for rigid lanthanide binding and paramagnetic NMR spectroscopy. AB - Attachment of two nitrilotriacetic acid-based ligands to a protein alpha-helix in an i, i + 4 configuration produces an octadentate chelating motif that is able to bind paramagnetic lanthanide ions rigidly and with high affinity, leading to large pseudocontact shifts and residual dipolar couplings in the NMR spectrum. PMID- 21617819 TI - Contrasting the denaturing effect of guanidinium chloride with the stabilizing effect of guanidinium sulfate. AB - Guanidinium chloride, GdmCl, is a strong denaturing agent of globular proteins, whereas guanidinium sulfate, Gdm(2)SO(4), is a stabilizing agent of globular proteins. The stabilizing activity of Gdm(2)SO(4) is unexpected because the denaturant capability of GdmCl is due to direct interactions of Gdm(+) ions with protein surface groups. It is shown that the statistical thermodynamic approach devised to explain the molecular origin of cold denaturation [G. Graziano, Phys. Chem. Chem. Phys., 2010, 12, 14245-14252] can provide a rationalization of the different behaviour of GdmCl and Gdm(2)SO(4) towards globular proteins. The fundamental quantity is the reversible work to create in the aqueous solution a cavity suitable to host the D-state and a cavity suitable to host the N-state. In aqueous GdmCl solutions, this contribution is not large enough to overwhelm the conformational entropy gain upon unfolding and the direct attractions between Gdm(+) ions and protein surface groups; in aqueous Gdm(2)SO(4) solutions, it is so large that it overwhelms the two destabilizing contributions. Sulfate ions, due to their high charge density, interact strongly with water molecules producing a number density increase, that, in turn, renders the cavity creation process very costly, reversing the denaturing power of Gdm(+) ions and stabilizing the N-state of globular proteins. PMID- 21617820 TI - Non aggregated colloidal silver nanoparticles for surface enhanced resonance Raman spectroscopy. AB - Silver nanoparticles with a tuneable lambda max were produced as colloids by heterogeneous nucleation. The synthesis process is both fast and repeatable, producing stable PVA capped nanoparticles. The colloid's effectiveness in the SERRS system was investigated using Rhodamine 6G, R6G, Crystal Violet, CV, and Malachite Green, MG, as probe molecules. A clear sensing trend was observed, where the Raman signal emitted was significantly enhanced by the addition of silver nanoparticles. A build up of signal intensity is observed until an optimum ratio is achieved, followed by a decline in signal intensity as the concentration of nanoparticles is further increased. The sensing trend appeared to be dependant on the structure of these model molecules with similarly structured compounds exhibiting similar trends. Thus a maximum enhancement with the Ag: analyte molar ratio of ~ 5.56: 1, was seen for CV and MG whereas R6G had a maximum enhancement at the Ag: analyte molar ratio of ~ 2.25: 1. PMID- 21617821 TI - Rapid screening swine foot-and-mouth disease virus using micro-ELISA system. AB - In order to tackle both regional and global foot-and-mouth disease virus (FMDV) epdimics, we hereby develop a rapid microfluidic thermal lens microscopic method to screen swine type O FMDV with good efficiency. The scheme has great merits in terms of field portability, sample volume, assay time, analytical sensitivity, and test reproducibility. PMID- 21617822 TI - Chemically resistant microfluidic valves from Viton(r) membranes bonded to COC and PMMA. AB - We present a reliable technique for irreversibly bonding chemically inert Viton(r) membranes to PMMA and COC substrates to produce microfluidic devices with integrated elastomeric structures. Viton(r) is widely used in commercially available valves and has several advantages when compared to other elastomeric membranes currently utilised in microfluidic valves (e.g. PDMS), such as high solvent resistance, low porosity and high temperature tolerance. The bond strength was sufficient to withstand a fluid pressure of 400 kPa (PMMA/Viton(r)) and 310 kPa (COC/Viton(r)) before leakage or burst failure, which is sufficient for most microfluidic applications. We demonstrate and characterise on-chip pneumatic Viton(r) microvalves on PMMA and COC substrates. We also provide a detailed method for bonding fluorinated Viton(r) elastomer, a highly chemically compatible material, to PMMA and COC polymers. This allows the production of microfluidic devices able to handle a wide range of chemically harsh fluids and broadens the scope of the microfluidic platform concept. PMID- 21617823 TI - Early development drug formulation on a chip: fabrication of nanoparticles using a microfluidic spray dryer. AB - Early development drug formulation is exacerbated by increasingly poor bioavailability of potential candidates. Prevention of attrition due to formulation problems necessitates physicochemical analysis and formulation studies at a very early stage during development, where the availability of a new substance is limited to small quantities, thus impeding extensive experiments. Miniaturization of common formulation processes is a strategy to overcome those limitations. We present a versatile technique for fabricating drug nanoformulations using a microfluidic spray dryer. Nanoparticles are formed by evaporative precipitation of the drug-loaded spray in air at room temperature. Using danazol as a model drug, amorphous nanoparticles of 20-60 nm in diameter are prepared with a narrow size distribution. We design the device with a geometry that allows the injection of two separate solvent streams, thus enabling co-spray drying of two substances for the production of drug co-precipitates with tailor-made composition for optimization of therapeutic efficiency. PMID- 21617824 TI - Triggered release of therapeutic antibodies from nanodiamond complexes. AB - Recent reports have revealed that detonation nanodiamonds (NDs) can serve as efficient, biocompatible, and versatile drug delivery platforms. Consequently, further investigations exploring additional therapeutic applications are warranted. Current limitations associated with the non-specific nature of intravenous drugs limit the potential of certain pharmacological agents. One such treatment that could benefit from a stable delivery platform is antibody (Ab) therapy. Determination of Ab adsorption and desorption to a ND surface was subsequently examined using the transforming growth factor beta (TGF-beta) antibody as a model therapeutic. ND-Ab complexes were found to be stable in water through enzyme-linked immunosorbent assays (ELISAs), UV-vis spectroscopy and TEM, with no Ab released after ten days. Released Abs were detected in extreme pH solutions (3.5), DMEM (+) serum with pH levels ranging from 4 to 10.5, and inorganic saline solutions. Preserved activity of Abs released in DMEM (+) serum was confirmed using an ELISA. These results suggest ND-Ab complexes are synthesized and stabilized in water and are triggered to release active Abs upon exposure to physiological conditions. PMID- 21617826 TI - Human papillomavirus genotypes among infected Thai women with different cytological findings by analysis of E1 genes. AB - Cervical cytological data may not be sufficient for cervical cancer screening and prevention. In this project, we determined HPV genotype among infected Thai women with different cytological findings by characterization of E1 genes. Five hundred and thirty-five specimens were tested by PCR amplification of the E1 genes. HPV genotypes were determined by sequencing, comparison with the GenBank database and were analyzed in relation to different cytological findings. HPV-DNA by PCR were typed and revealed 32 different genotypes. HR-HPV (HPV16, 18 or 52) was detected in all samples with cervical cancer cytology. HPV16 was most prevalent irrespective of cervical cytology. Moreover, HPV31 and 52 were most prevalent in the HSIL and LSIL groups whereas HPV66 was found mostly in the LSIL group. The LSIL group displayed the highest variation of HPV genotypes. Moreover, HPV31 and 52 predominated in the HSIL and LSIL groups especially HPV52 which was found in cancer samples. We hoped that these data of HPV genotypes can be used as preliminary data of HPV in Thailand and can serve as basic data for future research into the HPV genotype in south-east Asia. PMID- 21617825 TI - Italian guidelines for the use of antiretroviral agents and the diagnostic clinical management of HIV-1 infected persons. PMID- 21617827 TI - Comparison of real-time PCR and pp65 antigen assays for monitoring the development of Cytomegalovirus disease in recipients of solid organ and bone marrow transplants. AB - Cytomegalovirus (CMV) infection is a frequent complication in transplant recipients. This retrospective study compared real-time PCR (rt-PCR) and a pp65 antigen assay as tools for monitoring CMV infection in solid organ (SOT) and bone marrow (SCT) transplant patients. The study tested 2662 samples by rt-PCR, and 1284 specimens with a pp65 antigen assay. 24.3% of the rt-PCR samples and 4.1% of the pp65 antigen samples were positive. 793 specimens, from 230 patients, were tested with both assays. In 6.7% of samples, both tests were positive; in 72.7% both were negative; in the remaining 20.6% of cases, the results were discordant. CMV disease was diagnosed in 50 patients. Results from the two methods were poorly correlated (r=0.460). The sensitivity of rt-PCR (94%) was higher than that of the pp65 antigen assay (27%). Both assays showed high specificity (92% and 99%, respectively). ROC curve analysis, performed separately for SOT and SCT patients, confirmed that rt-PCR outperformed the pp65 assay in the detection of CMV. These findings provide evidence that rt-PCR is a reliable diagnostic tool, and that it can be more effective than pp65 based assays in monitoring CMV infection progression and in guiding therapy in immunocompromised patients. PMID- 21617828 TI - Quantitative viral load measurement for BKV infection in renal transplant recipients as a predictive tool for BKVAN. AB - Infection by polyomavirus BK (BKV) is an emerging problem in the clinical management of renal transplant patients because it is responsible for nephropathy and consequently can cause loss of the transplanted organ (BKV associated nephropathy, BKVAN). Aim of this study was to evaluate the use of blood viral load measurement as a screening tool for diagnosis of BKV infection and to identify a threshold value for the management of patients. A total of 75 kidney transplant patients, corresponding to 338 consecutive plasma samples, were analyzed by an automatic system for nucleic acid extraction and quantitative real time polymerase chain reaction (PCR) for detection of BKV. BKV was detected in 170 samples (26 patients) with a median viral load of 4.1 log10 copies/mL; among these 26 patients, seven (34.7%) were found to have BKVAN on allograft biopsy together with a median viral load of 5 log10 copies/mL. The ROC curve analysis identified a viral load equal to 4.1 log10 copies/mL as the best discriminant cut off value to predict the disease and to identify patients at risk of developing BKVAN. PMID- 21617830 TI - Microbiological quality of Pecorino Siciliano "primosale" cheese on retail sale in the street markets of Palermo, Italy. AB - Pecorino Siciliano (PS) "primosale" is a traditional Sicilian fresh soft cheese made from sheep's milk. Short-ripening time and production from unpasteurized or raw milk can facilitate bacterial contamination of PS "primosale". The microbiological quality of "primosale" on retail sale in the street markets of Palermo, Italy was studied by detecting the common food pathogens Listeria monocytogenes and Staphylococcus aureus and indicator microorganisms, such as Escherichia coli, Enterobacteriaceae and Staphylococcaceae. In our study, 4% and 44% of the samples, respectively, did not comply with the acceptability levels fixed by European regulations for S. aureus and E. coli. A high contamination of bacteria belonging to Enterobacteriaceae and Staphylococcaceae was found in 42% and 50% of the cheeses analyzed, respectively. Such results indicate poor husbandry and poor hygiene practices during milk collection or preservation or during cheese production processes and handling. In addition, the retail sale conditions may have played a role in cheese contamination since a correlation was found between poor microbiological quality and some selling parameters. This study emphasizes the need to improve production hygiene throughout the PS food chain in line with the traditional cheese-making procedures. Labelling of PS with clear information on whether the cheese was prepared from raw milk also requires improvement. PMID- 21617829 TI - The diagnostic value of Western blot method in patients with cystic echinococcosis. AB - Cystic echinococcosis (CE) is the larval cystic stage (called echinococcal cysts) of a small taeniid-type tapeworm (Echinococcus granulosus). Carnivores such as dogs are usually definitive hosts. Intermediate hosts are typically herbivores such as sheep and cattle. CE can be detected using various imaging techniques such as ultrasonography or radiology. Moreover the primary diagnosis has to be confirmed by serological tests since the clinical signs of the disease are non specific. This study examined the antigenic band patterns useful for serologic diagnosis of hydatidosis. We also report on the post-operative evolution of patients treated for this disease and also determined the diagnostic performance of Western blot IgG kit. Twenty-five (16 females and 9 males) non-operated patients with hydatid cysts (NOP) and 33 (21 females and 12 males) operated patients with hydatid cysts (OP) were included as study group and 22 healthy individuals (14 females and 8 males) with no known chronic diseases were included as a control group. The ages of the patients and control group individuals were between 16-83 years. Patient and control groups were matched for age and sex. Cyst hydatid IgG antibodies were detected in the sera from all patient groups but no antibodies were found in the sera from the control group using ELISA IgG method. Twenty-three (92%) non-operated patients and 18 (54.5%) operated patients exhibited positive results when Western blot IgG kit was used. The P7 band pattern was detected in the sera from all operated and non-operated patients. Twenty-seven of these positive cases had p7 and (p7+p16/18), (p7+p24/26) or (p7+p16/18+p24/26). No antibodies against p7, p16/18 ve p24/26 band patterns were seen in sera from the control group A statistically significant difference was detected between operated and nonoperated patients for Western blot positivity.(p<0.01). p: 0.018- X2=5,604- OR: 0.176- 95% CI: 0.037- 0.841. The sensitivity, specificity, positive prediction and negative prediction values of Echinococcus granulosus Western blot kit for 25 cases with CE and 22 healthy controls were calculated as 92%, 100%, 100% and 91.7%, respectively. In conclusion, we suggest that monitoring p7 in all non-operated patients may be useful to determine the efficiacy of medical treatment and that monitoring p7 antibodies using serological and Western blot methods in operated patients may be useful for the screening of post-operative evolution in patients with hydatid cyst. PMID- 21617831 TI - Erythromycin and penicillin resistance mechanisms among viridans group streptococci isolated from blood cultures of adult patients with underlying diseases. AB - The aim of the study was to evaluate the species distribution, antimicrobial susceptibility and erythromycin-penicillin resistance mechanisms of viridans streptococci (VGS) isolates from blood cultures of adult patients with underlying diseases. Fifty VGS blood culture isolates were screened for their antibiotic susceptibilities against penicillin G, erythromycin and tetracycline by E-test. Clindamycin, cefotaxime, chloramphenicol, levofloxacin, linezolid and vancomycin susceptibility were performed by broth microdilution method. Erythromycin and penicillin resistance genotypes, ermB and mefA/E, pbp1a, pbp2b and pbp2x are amplified using PCR method. The clinical isolates included Streptococcus mitis (n. 19), S.oralis (n. 13), S.sanguinis, S.parasanguinis (n. 6, each), S.salivarius, S.vestibularis (n. 2, each), S.constellatus, S.sobrinus (n. 1, each). The percentage resistance against erythromycin and penicillin was 36% and 30%, respectively. The genotypic carriage rate of erythromycin resistance genes were: 56% ermB, 28% mefE, 8% ermB+mefE. Penicillin-resistant isolates carried pbp2b (33.3%) and pbp2x (20%) genes. Twenty-four VGS isolates were recovered from patients with cancer. S.mitis and S.oralis predominated among patients with cancer who had erythromycin and penicillin resistance isolates. The importance of classical antimicrobial agents like penicillin and erythromycin warrants the continuous surveillance of invasive VGS isolates and can guide better treatment options especially in patients with underlying diseases. PMID- 21617832 TI - Interlaboratory evaluation of VITEK2 system and Sensititre YeastOne(r) for antifungal susceptibility testing of yeasts isolated from blood cultures against four antifungal agents. AB - An interlaboratory evaluation (seven centers) of VITEK2 System and Sensititre YeastOne(r) was conducted to test the antifungal susceptibilities of yeasts. The MICs of amphotericin B, fluconazole, flucytosine, and voriconazole were determined for 70 isolates of Candida spp. Our results demonstrated a higher interlaboratory agreement of VITEK 2 System than Sensititre YeastOne(c). A good concordance between the two methods was observed for amphotericin B, fluconazole, voriconazole and 5-fluorocytosine (from 81.4% to 88.6%). The study suggests the potential value of the VITEK2 System as a convenient alternative method for testing the susceptibility of yeasts. It also indicates the need for further optimization of MIC endpoint criteria to improve interlaboratory agreement. PMID- 21617833 TI - Toxoplasmosis in Kosovo pregnant women. AB - This study presents the initial results of a collaborative project aimed at the evaluation of Toxoplasma seroprevalence in a population of Kosovar pregnant women. The serum samples of 334 pregnant women were tested to detect IgG, IgM, IgG avidity for toxoplasmosis. Data regarding age, occupation, area of origin and education were also obtained for the pregnant women examined; 97/334 (29.4%) resulted positive for IgG antibodies, four of whom (4.1%) were also positive for IgM, (1.2% of the total population examined). All four IgM-positive pregnant women also demonstrated low avidity tests. The rate of IgG seroprevalence found in our study was lower than that observed in various European countries, especially those of western Europe. Conversely, the percentage of recent infections was higher than expected. The higher rate of infections could be the result of a recent toxoplasmosis epidemic in Kosovo, most likely due to the altered hygienic conditions caused by the forced transfer of the ethnic-Albanian population from an area of low (Serbia) to high (Kosovo) toxoplasmosis prevalence. PMID- 21617834 TI - Transmission electron microscopy as a tool for exploring bacterial proteins: model of RickA in Rickettsia conorii. AB - Rickettsia conorii, the etiologic agent of Mediterranean spotted fever, belongs to the spotted fever group of Rickettsia. It is an obligate intracellular bacterium that grows within the cytoplasm of its eukaryotic host cells. It is motile in the cytoplasm of infected cells and RickA is reported as critical protein in this aspect. However, the subcellular localization of RickA remains uncertain. We describe a simple method allowing RickA protein to be localized by immunofluorescence assay (IFA) and transmission electron microscopy (TEM). By using IFA we showed the global expression of surface protein RickA in R. conorii organisms. The TEM results showed that RickA is widely expressed over the entire bacterial surface of R. conorii. PMID- 21617835 TI - Frequency of seropositivity for Coxiella burnetii immunoglobulins in livestock and abattoir workers in Trinidad. AB - Coxiella burnetii is a zoonotic, rickettsial pathogen which causes mild and severe diseases often referred to as Q-fever in humans, particularly those occupationally exposed. This study determined the seropositivity for Coxiella burnetii IgM immunoglobulins using the enzyme immunoassay (EIA) in livestock and abattoir workers in Trinidad and related to selected personal characteristics to seroprevalence. Overall, of the 455 humans whose serum samples were tested, 20 (4.4%) were seropositive for C. burnetii IgM immunoglobulin, comprising 13 (4.6%) out of 283 livestock workers, 4 (4.7%) out of 85 abattoir workers and 3 (3.4%) out of 87 office workers (P>0.05; Chi 2). The age, sex and race of workers were not significantly associated with the occurrence of acute Q-fever (P>0.05; Chi 2). This is considered the first documentation of Q-fever in the human population in Trinidad. It is difficult to assess the impact of the disease in the country since the disease is not routinely tested for in the local hospitals or diagnostic laboratories. PMID- 21617836 TI - Co-expression of plasmid-mediated quinolone resistance-qnrA1 and blaVEB-1 gene in a Providencia stuartii strain. AB - An extended-spectrum B-lactamase (ESBL)-producing Providencia stuartii isolate was studied. A qnrA1 gene co-expressing blaVEB-1 gene was detected. Both genes were transferred to the recipient strain. The ciprofloxacin MIC of recipient strain increased tenfold. The blaVEB-1 gene persisted in microorganisms in Turkey but it also spread with PMQR genes to other species. The combination of PMQR with multidrug resistant isolates producing ESBLs may compromise the use of valuable antibiotics. Serious efforts are necessary to detect PMQR determinants not only with common B-lactamases in widespread pathogens but also with uncommon forms that are encountered infrequently. PMID- 21617837 TI - Effect of brief daily exercise on headache among adults--secondary analysis of a randomized controlled trial. AB - OBJECTIVE: This paper investigates secondary outcomes (headache) in a randomized controlled trial with physical exercise among office workers with neck/shoulder pain. METHODS: A total of 198 office workers with frequent neck/shoulder pain were randomly allocated to either one of two intervention groups (10 weeks of resistance training with elastic tubing for 2 or 12 minutes per day, 5 times a week) or the control group, which received weekly health information. Secondary outcomes included changes in frequency, intensity, and duration of headache after ten weeks. RESULTS: Compared with the control group, headache frequency decreased in the 2- and 12-minute intervention groups [0.64 days/week (95% confidence interval [95% CI]) 0.23-1.0) and 0.79 days/week (95% CI 0.37-1.2), corresponding to a 43% and 56% decrease from baseline, respectively]. Intensity and duration of the remaining headaches were unaffected. CONCLUSIONS: Two minutes of daily resistance training for ten weeks reduces headache frequency among office workers with neck/shoulder pain. The vast number of adult workers suffering from one or two days of weekly headaches and who could potentially comply with and benefit from brief exercise programs stresses the applicability of our findings. PMID- 21617838 TI - Identification of biological functions and gene networks regulated by heat stress in U937 human lymphoma cells. AB - Although cancer cells exposed to temperatures >42.5 degrees C undergo cell death as the temperature rises, exposure of up to 42.5 degrees C induces slight or no cytotoxicity. The temperature of 42.5 degrees C is, therefore, well known to be the inflection point of hyperthermia. To better understand the molecular mechanisms underlying cellular responses to heat stress at temperatures higher and lower than the inflection point, we carried out global scale microarray and computational gene expression analyses. Human leukemia U937 cells were incubated at 42 degrees C or 44 degrees C for 15 min and cultured at 37 degrees C for 0-6 h. Apoptosis accompanied by the activation of caspase-3 and DNA fragmentation was only observed in cells treated with heat stress at 44 degrees C, but not at 42 degrees C. Although a large number of genes were differentially expressed by a factor of 2.0 or greater, we found substantial differences with respect to the biological functions and gene networks of the genes differentially expressed at the two temperatures examined. Interestingly, we identified temperature-specific gene networks that were considered to be mainly associated with cell death or cellular compromise and cellular function and maintenance at 44 degrees C or 42 degrees C, respectively, by using the Ingenuity pathway analysis tools. These findings provide the molecular basis for a further understanding of the mechanisms of the biological changes that are responsive to heat stress in human lymphoma cells. PMID- 21617839 TI - Phase II study of the antiretroviral activity and safety of the glucocorticoid receptor antagonist mifepristone in HIV-1-infected patients. AB - Preclinical studies have shown that the anti-glucocorticoid drug mifepristone effectively inhibits HIV replication both in vitro and in vivo. However, the drug did not demonstrate anti-HIV activity in a previous phase I/II study when administered at the daily dose of 75-225 mg. The aim of this study was to assess whether mifepristone may exert antiretroviral activity or influence immunological parameters when administered orally at daily doses of 150 or 300 mg in highly active antiretroviral therapy (HAART)-naive HIV-infected patients. We performed an open label non-randomized phase II study that included 26 patients who underwent 28 days of once daily oral administration of 150 (12 subjects) or 300 mg (14 subjects) of mifepristone. A total of 3 patients dropped out of the study, respectively 1 in the 150 mg dose group and 2 in the 300 mg dose group. The main hemato-chemical alterations reported were hypokalemia and increase in the blood levels of cortisol, especially in those patients that received mifepristone at the dose of 300 mg/day. Although we observed a trend of reduced viral load along the study in both groups, statistical significance was not achieved for either the primary nor the secondary endpoints. In summary, mifepristone treatment was well-tolerated but it failed to significantly influence viro-immunological parameters in HAART-naive HIV-infected patients. PMID- 21617840 TI - Inhibition of glypican-3 expression via RNA interference influences the growth and invasive ability of the MHCC97-H human hepatocellular carcinoma cell line. AB - Glypican-3 (GPC3), a membrane-bound heparan sulfate proteoglycan, is found to be overexpressed in hepatocellular carcinoma (HCC). The purpose of the present study was to investigate the possible role of GPC3 in the development of HCC. In this study, RNA interference (RNAi) with a GPC3 small hairpin RNA (GPC3 shRNA) was used to identify the effects of GPC3 on the regulation of malignant behaviors of HCC. MHCC97-H, a highly metastatic human HCC cell line in which GPC3 mRNA and protein levels were detected as the highest among the 4 HCC cell lines assessed in this study, and was thus selected as a cell model for in vitro and in vivo experiments. The results showed that down-regulation of GPC3 can significantly inhibit the proliferative and invasive ability of MHCC97-H. Compared with the parental HCC cells, GPC3-silenced cells exhibited attenuated capacities in developing tumors in nude mice, while the growth of tumor xenografts derived from these cells dramatically regressed. In conclusion, our results suggest that GPC3 contributes to the proliferation and metastasis of HCC and that it may be a potential molecular target for HCC treatment. PMID- 21617841 TI - State-of-the-art bioinformatics protein structure prediction tools (Review). AB - Knowledge of the native structure of a protein could provide an understanding of the molecular basis of its function. However, in the postgenomics era, there is a growing gap between proteins with experimentally determined structures and proteins without known structures. To deal with the overwhelming data, a collection of automated methods as bioinformatics tools which determine the structure of a protein from its amino acid sequence have emerged. The aim of this paper is to provide the experimental biologists with a set of cutting-edge, carefully evaluated, user-friendly computational tools for protein structure prediction that would be helpful for the interpretation of their results and the rational design of new experiments. PMID- 21617842 TI - Molecular modulation of osteocalcin and its relevance in diabetes (Review). AB - Osteocalcin (OC) is encoded by the bone gamma-carboxyglutamate (Gla) protein (BGLAP) gene, and it is released by osteoblasts during osteogenesis. Its expression can be modulated by growth factors, hormones, cytokines and physical stimuli via signal transduction pathways, binding to the BGLAP gene promoter or interactions with nuclear transcription factors. It was recently demonstrated that uncarboxylated OC improves glucose tolerance and insulin sensitivity in mice by increasing the expression and secretion of insulin in beta-cells and of adiponectin in adipocytes. Humans with type 2 diabetes have significantly lower serum levels of OC than healthy individuals and indeed, serum OC levels have been inversely correlated with fasting plasma glucose, fasting insulin and the homeostasis model assessment of insulin resistance (HOMA-IR) index. Moreover, several drugs have been shown to influence OC expression and its serum concentration. This review summarizes the molecular mechanisms involved in the modulation of OC expression, and discusses the potential relevance of OC in the pathogenesis and treatment of diabetes. PMID- 21617843 TI - Biocompatibility of magnesium-zinc alloy in biodegradable orthopedic implants. AB - In this study, magnesium-zinc (Mg-Zn) alloy was investigated as a biodegradable orthopedic implant. MC3T3-E1 cell attachment, mineralization and osteogenic specific mRNA expression were assessed for as measurements of the in vitro biocompatibility of Mg-Zn alloy. In vivo degradation of the Mg-Zn alloy and the accompanying new bone formation in the femoral marrow cavity were analyzed by scanning electron microscopy and histomorphological analysis. Results showed that MC3T3-E1 cells cultured on Mg-Zn alloy samples manifested better attachment and mineralization ability, as well as improved mRNA expression of collagen 1 alpha 1 (COL1alpha1) and osteocalcin (OC), compared with cells seeded on poly-L-lactic acid (PLLA) samples. In vivo experiments demonstrated that, compared with PLLA materials, the Mg-Zn alloy not only degraded faster, but was accompanied by considerable new bone formation around the samples. Our data indicate that Mg-Zn alloy has excellent biocompatibility for application as degradable bone implants. PMID- 21617844 TI - Increased expression of alphaA-crystallin in human diabetic eye. AB - We recently demonstrated that alphaA-crystallin, a molecular chaperone, protected photoreceptors from apoptotic signals in intraocular inflammation. Advanced glycation end product (AGE) plays an important role in the progression of diabetic retinopathy. The aim of this study was to examine the expression of alpha-crystallins and apoptosis in human diabetic retina, and to analyze alpha crystallin up-regulation in murine eyes after AGE stimulation. Eight eye globes were obtained from postmortem donors. Six out of the eight had a medical history of diabetes mellitus, while two were without diabetes. Formalin-fixed, paraffin embedded tissue sections were subjected to H&E staining and immunohistochemistry with anti-alphaA and alphaB-crystallins, anti-AGE and receptor for AGE (RAGE) antibodies. Apoptotic cells were detected by the TUNEL assay. Recombinant AGE protein was injected into the vitreous of adult murine eyes, and the posterior eyecups were excised 4 days after the administration. Western blot analyses and quantitative real-time PCR were performed to evaluate the alteration of alpha crystallin expression. Histopathology revealed no remarkable differences between diabetic and non-diabetic retinas. Immunoreactivity for alphaA-crystallin was predominantly detected in the diabetic retina, whereas alphaB-crystallin expression was relatively low. AGE immunoreactivity was highly detected in diabetic retina and the vitreous, whilst immunoreactivity for RAGE was less marked. TUNEL-positive apoptotic cells were occasionally observed in photoreceptors of the diabetic retina, whereas cytoplasmic immunoreactivity for alphaA-crystallin was relatively low. alphaA-crystallin expression was up regulated, and alphaB-crystallin was down-regulated in murine posterior eyecups exposed to AGE protein. The mRNA levels of alphaA-crystallin were significantly up-regulated, whereas those of alphaB-crystallin remained unchanged after AGE stimulation. Thus, alphaA-crystallin and AGE were highly expressed in human diabetic retina. alphaA-crystallin responded to AGE accumulation, which may contribute to the protection of photoreceptors against AGE-related retinal tissue injury. PMID- 21617845 TI - Poly(ADP-ribose)polymerase inhibition counteracts renal hypertrophy and multiple manifestations of peripheral neuropathy in diabetic Akita mice. AB - Poly(ADP-ribose) polymerase (PARP) activation has been implicated in the pathogenesis of diabetic complications, including nephropathy and peripheral neuropathy. This study aimed at evaluating the manifestations of both complications in diabetic Akita mice, a model of Type 1 (insulin-dependent) diabetes, and their amenability to treatment with the potent PARP inhibitor, 10 (4-methyl-piperazin-1-ylmethyl)-2H-7-oxa-1,2-diaza-benzo[de] anthracen-3-one (GPI 15427). Male non-diabetic C57Bl6/J and diabetic C57Bl/6-Ins2Akita/J (Akita) mice were maintained with or without treatment with GPI-15427, 30 mg/kg/day, for 4 weeks starting from 16 weeks of age. Sixteen week-old Akita mice displayed sensory nerve conduction velocity (SNCV) deficit, whereas the motor nerve conduction velocity (MNCV) tended to decrease, but the difference with controls did not achieve statistical significance. They also developed thermal and mechanical hypoalgesia and tactile allodynia. SNCV deficit, mechanical hypoalgesia, and tactile allodynia progressed with age whereas the severity of thermal hypoalgesia was similar in 16- and 20-week-old Akita mice. PARP inhibition alleviated, although it did not completely reverse, SNCV deficit, thermal and mechanical hypoalgesia and tactile allodynia. Sixteen-week-old Akita mice displayed MNCV deficit (41.3+/-2.5 vs. 51.0+/-1.2 m/sec in non-diabetic controls, P<0.01), axonal atrophy of myelinated fibers, kidney hypertrophy, and albuminuria. MNCV slowing, axonal atrophy, and kidney hypertrophy, but not albuminuria, were less severe in GPI-15427-treated age-matched Akita mice. Neuroprotective and nephroprotective effects of PARP inhibition were not due to alleviation of diabetic hyperglycemia, or peripheral nerve p38 mitogen-activated protein kinase activation. GPI-15427 did not affect any variables in control mice. In conclusion, the findings support an important role for PARP activation in diabetic peripheral neuropathy and kidney hypertrophy associated with Type 1 diabetes, and provide rationale for development and further studies of PARP inhibitors, for the prevention and treatment of these complications. PMID- 21617846 TI - Ghrelin reduces rat myocardial calcification induced by nicotine and vitamin D3 in vivo. AB - Ghrelin, a newly discovered bioactive peptide, initially was identified as a strong stimulant for the release of growth hormone (GH) and that has improved cardiac function in patients suffering from end-stage chronic heart failure. Increasing evidence has demonstrated that ghrelin may have myocardial protective effects. However, the role of ghrelin in the pathogenesis of cardiovascular diseases remains unclear. In this study, an in vivo model of rat myocardial calcification induced by vitamin D3 and nicotine was used to study the possible mechanism in the regulatory action of ghrelin on the calcified myocardium. Calcification increased total Ca2+ content and 45Ca2+ deposition in the myocardium and alkaline phosphatase (ALP) activation in the plasma. Compared with the control group, ghrelin mRNA expression was up-regulated and the myocardium calcium content was significantly increased in vitamin D3 and nicotine-treated rats. Rats were subcutaneously injected with 1 or 10 nmol/kg ghrelin. Rats treated with both low- and high-dose ghrelin decreased total Ca2+ content and 45Ca2+ deposition in cardiac muscle and inhibited ALP activation in the myocardium and plasma, in a concentration-dependent manner. In addition, osteopontin (OPN) mRNA expression significantly decreased and that of endothelin (ET-1) significantly increased with myocardial calcification. Ghrelin treatment increased OPN expression at the mRNA level and reduced ET-1 mRNA expression in a dose-dependent manner. These results indicate that exogenous administration with ghrelin attenuates myocardial calcification induced by nicotine and vitamin D3, and that the possible mechanism is via the ghrelin-induced increase in the OPN mRNA levels and decrease in the ET-1 mRNA expression in the myocardium. PMID- 21617847 TI - Leptin gene transfer regulates fibromuscular development and lipid deposition in muscles via SIRT1, FOXO3a and PGC-1alpha in mice in vivo. AB - Leptin gene transfer in the liver by hydrodynamic-based gene delivery instead of peptide administration was used to investigate the effects of leptin on muscle mass accretion and lipid accumulation in muscles of wild-type mice. Food intake (P<0.01), body weight (P<0.01) and white adipose tissue (P<0.01) were significantly reduced in the leptin gene-treated group compared with the control group. Moreover, plasma leptin concentration was significantly increased after administration of the mouse leptin gene at a dose of 15 ug per mouse for 1 day (P<0.01) or 1 week (P<0.05). Furthermore, the mRNA abundance of myosin heavy chain type I (MyHC-I), myosin heavy chain type II (MyHC-IIa, MyHC-IIx), adipose triglyceride lipase (ATGL), hormone-sensitive lipase (HSL) and lipoprotein lipase (LPL) genes in gastrocnemius muscle and extensor digitorum longus after administration of leptin for 1 week were significantly increased compared with the control group. Finally, we investigated the mechanism by which leptin gene transfer affects fibromuscular and fat deposition in muscle. Gene expression and protein levels of SIRT1, and proliferator-activated receptor-gamma coactivator 1alpha (PGC-1alpha) were remarkably increased in extensor digitorum longus. On the other hand, PGC-1alpha and FOXO3a gene expression was observed to have significantly increased in gastrocnemius muscle. However, only changes in the protein levels of PGC-1alpha were observed (P<0.05). These results suggest that leptin may affect the growth and development of muscle, and fat deposition in wild-type mice via SIRT1 and FOXO3a and their downstream targets, including PGC 1alpha. PMID- 21617848 TI - Role of periostin in cancer progression and metastasis: inhibition of breast cancer progression and metastasis by anti-periostin antibody in a murine model. AB - Periostin (PN), a secreted adhesion-related protein expressed in the periosteum and periodontal ligaments, acts as a critical regulator of the formation and maintenance of bone and teeth, and also plays an important role in tumorigenesis. Although PN is highly expressed in various types of human cancers, its function is still unclear. In this study, we focused on the exon 17 region of PN, which is alternatively spliced out. To investigate the function of full-length PN with exon 17, we produced a neutralizing antibody (PN1-Ab) against the peptide encoded by exon 17. In vivo, administration of PN1-Ab significantly inhibited the growth of primary tumors as well as metastatic tumors, associated with prevention of bone destruction, resulting in increased survival of mice. Consistent with in vivo data, the present in vitro study demonstrated that addition of full-length PN significantly inhibited cell adhesion and detached adherent cells, while PN1 Ab inhibited the action of PN in a dose-dependent manner. In addition, PN1-Ab significantly inhibited the proliferation, migration and invasion of 4T1 mouse breast cancer cells, which produced PN. Interestingly, PN1-Ab also inhibited the differentiation of osteoclasts. Overall, the present study demonstrated that PN plays a pivotal role in the progression and metastasis of breast cancer. Since administration of PN1-Ab prolonged cell survival through inhibition of the progression and metastasis of 4T1 cells, further development of the PN1-Ab such as generation of a humanized antibody may provide a new therapeutic agent against breast cancer. PMID- 21617849 TI - Aspirin reduces the apoptotic effect of etoposide via Akt activation and up regulation of p21(cip). AB - Previous studies on the apoptotic effect of aspirin mainly focus on colorectal cancer and breast carcinoma. Few studies have been designed to explore the effect of aspirin on hepatocellular carcinoma. In the present study, we observed that aspirin caused G0/G1 phase cell cycle arrest and reduced etoposide induced caspase-3 activation in hepatocellular carcinoma G2 (HepG2) cells. Further investigation demonstrated that aspirin notably enhanced the activity of Akt and ERK1/2. Blocking the activation of Akt by the PI3-K-selective inhibitor wortmannin abrogated the anti-apoptotic effect of aspirin while the MEK inhibitor U0126 did not. p21(cip), an important substrate of Akt, is involved in the regulation of cell cycle arrest and apoptosis. Our data showed that the protein expression and ser146 phosphorylation levels of p21(cip) were significantly increased after treatment with aspirin, whereas p53 or p27 showed no change. The increase of p21(cip) protein levels was also scavenged by wortmannin but not by U0126. Moreover, reduction of caspase-3 activity induced by aspirin was attenuated by silencing p21(cip) expression. These results indicated that the anti-apoptotic effect of aspirin was dependent on activation of Akt which inhibited cell apoptosis by up-regulating p21(cip) and blocking caspase-3 activation. These findings could have clinical relevance in anticancer therapy and aspirin co-treatment of human malignancies. PMID- 21617850 TI - Dickkopf-1 in human oral cancer. AB - Dickkopf-1 (Dkk1), a negative regulator of the Wnt signaling pathway, is implicated in tumorigenesis in several types of cancer. The purpose of this study was to determine the involvement of Dkk1 in oral squamous cell carcinoma (OSCC). We found that Dkk1 is frequently upregulated in OSCC-derived cell lines and primary OSCCs compared with normal counterparts. Unexpectedly, Dkk1-positive cases were correlated significantly (P<0.05) with a low risk of regional lymph node metastasis. We also found that cellular migration and invasiveness increased in Dkk1 knockdown cells and decreased in Dkk1 overexpressed cells. Furthermore, we investigated the relationship between the expression of Dkk1 and distribution of beta-catenin in OSCC cells, since the Wnt signaling pathway is related closely to beta-catenin. Whereas alteration of the beta-catenin levels was not observed in each subcellular fractionation, the phosphorylated beta-catenin levels in nuclei increased in Dkk1 knockdown cells and decreased in Dkk1 overexpressed cells. These data indicated that the high phosphorylation level of beta-catenin in nuclei was correlated with a high risk of tumor invasiveness. The current study suggested that Dkk1 plays an important role in regulating cellular migration and invasiveness, making Dkk1 a potential biomarker for early detection of lymph node metastasis in OSCCs. PMID- 21617851 TI - Overexpression of EGR-1 modulates the activity of NF-kappaB and AP-1 in prostate carcinoma PC-3 and LNCaP cell lines. AB - To address elements that might uniquely characterize EGR-1 mediated signaling, the expression of two transcription factors, namely, nuclear factor kappa B (NF kappaB) and activator protein-1 (AP-1) were studied. PC-3 and LNCaP prostate carcinoma cell lines were transiently transfected with wild-type Egr-1 expression plasmid (pCMV-Egr-1) and treated with cisplatin and TPA. Overexpression of EGR-1 was found to induce nuclear expression of both, NF-kappaB and AP1. However, the intensity of the induced AP-1 and NF-kappaB was diminished after cisplatin treatment, but not after TPA. Our findings confirm that the overexpression of wild-type Egr-1 caused a marked increase in cell proliferation in PC-3 and LNCaP proliferation in a 14-day soft agar colony forming assay. In addition, luciferase reporter gene assay showed that the transcriptional activity of AP-1 and NF kappaB in PC-3 and LNCaP prostate carcinoma cell lines was also modulated by the overexpression of EGR-1 in these cells using tandem repeated Luc-AP-1 and Luc-NF kappaB. The activation of both NF-kappaB and AP-1 are key steps in the cascade of events following the activation of the EGR-1 gene. It was revealed that overexpression of EGR-1 selectively increased AP-1 and NF-kappaB activation, and that the activation of these nuclear factors appears to be essential for the induction of proliferation and anchorage independence in activated PC-3 and LNCaP cells. However, the mechanism underlying the modulation of AP-1 and NF-kappaB by the overexpression of EGR-1 is still unknown. PMID- 21617852 TI - Co-expression of laminin beta3 and gamma2 chains and epigenetic inactivation of laminin alpha3 chain in gastric cancer. AB - Laminin-332 (LM-332, formerly termed laminin-5) is a heterotrimeric glycoprotein that regulates cell adhesion and migration. Molecular alterations of LM-332 are involved in cancer progression. The aim of this study was to clarify alterations of LM-332 in gastric carcinoma. The expression of LM-332 subunits in 10 gastric carcinoma cell lines was investigated by RT-PCR, Western blotting, and immuno cytochemical/immunofluorescent analyses. The promoter methylation status of LM 332-encoding genes (LAMA3, LAMB3 and LAMC2) was analyzed by methylation-specific PCR (MSP). The relationship between cell migration and LM-332 expression was assessed by the scratch assay. The expression of LM-332 was analyzed immunohistochemically in 90 gastric cancer tissues. Co-expression of laminin beta3 and gamma2 chains was often observed in gastric carcinoma cell lines at mRNA and protein levels. In contrast, there was no expression of laminin alpha3 at either the mRNA or protein levels. Extra-cellular secretion of laminin beta3 and gamma2 chains was found in 2 of the 10 cell lines. The LAMA3 gene was transcriptionally silenced by methylation of the promoter CpG islands in all of the cell lines, while the LAMB3 and LAMC2 genes were silenced in several cell lines. Treatment with a demethylating agent, 5-aza-2'-deoxycytidine (5-aza-dC), restored expression of the LM-332-encoding genes. Methylation frequency of LAMA3 was higher than those of the LAMB2 and LAMC2 genes in gastric cancer tissues. Migration distances were significantly correlated with cytoplasmic laminin gamma2 chain expression. Immunohistochemistry showed frequent co-expression of laminin beta3 and gamma2 chains in gastric carcinoma cells, which was significantly correlated with depth of invasion and advanced tumor stage. The results suggest that the laminin beta3 and gamma2 chains accumulate intracellularly and play a role in gastric cancer progression, while epigenetic silencing of the laminin alpha3 chain may lead to inability to synthesize the basement membrane and may affect cancer cell invasion. Cancer cell motility appears to be associated with the cyto-plasmic laminin gamma2 chain in vitro. PMID- 21617853 TI - Aberrant DNA methylation of T-cell leukemia, homeobox 3 modulates cisplatin sensitivity in bladder cancer. AB - The development of resistance to cisplatin during treatment of bladder cancer constitutes a major obstacle to curing bladder cancer. The identification of epigenetic biomarker molecules for cisplatin resistance and the elucidation of the function of the identified genes in bladder cancer will provide useful information. We found that the candidate gene TLX3 was unmethylated in cisplatin sensitive cells and methylated in resistant cells. The suppression of TLX3 expression using TLX3-specific shRNA in parental cells increased cisplatin resistance. Contrarily, overexpression of TLX3 in resistant cells induced increased sensitivity to cisplatin. We found that 22 (21%) out of 110 clinical samples of bladder cancer showed the methylated pattern using the COBRA assay in TLX3. We found a correlation between TLX3 methylation and the sensitivity to cisplatin in the clinical samples by SDI test. Cisplatin sensitivity was closely associated with the methylation status of TLX3. These findings showed that the TLX3 methylation may be useful as a novel biomarker for cisplatin resistance and can be used to design therapies to counteract the resistance against cisplatin in bladder cancer. PMID- 21617854 TI - Promoter methylation of leukemia inhibitory factor receptor gene in colorectal carcinoma. AB - Aberrant methylation of gene promoters and corresponding loss of gene expression plays a critical role in the initiation and progression of colorectal cancer. An IL-6-type cytokine receptor, leukemia inhibitory factor receptor (LIFR), is a component of cell-surface receptor complexes for multifunctional cytokines such as LIF. Herein, we report that LIFR is methylated in human colon cancer. LIFR promoter was methylated in primary tumor tissues with high frequency (65%, 52/80). Quantitative methylation-specific PCR (TaqMan-MSP) demonstrated differential promoter methylation of LIFR in primary colorectal cancer tissues as compared to normal colon tissues (5%, 4/80). LIFR methylation was not detectable in 13 normal colon mucosa samples obtained from patients without cancer. The mRNA expression of LIFR was significantly down-regulated in colon cancer tissues as compared to corresponding normal tissues. A strong expression of LIFR protein was observed in all non-malignant normal and adjacent normal colon mucosa tissues whereas down-regulated LIFR protein expression was observed in primary tumors. These results demonstrate that cancer-specific methylation and a specific decrease of LIFR expression are a common inactivation event in colon cancer development. PMID- 21617855 TI - Caffeine activates tumor suppressor PTEN in sarcoma cells. AB - The tumor suppressor phosphatase and tensin homolog deleted on chromosome 10 (PTEN) is a negative regulator of the phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway. Akt activation exerts a strong anti-apoptotic effect and inhibits key pro-apoptotic proteins. We investigated the effect of caffeine in the prevention of tumor cell proliferation and induction of cell death. We found that caffeine induced increased intracellular cAMP levels, PTEN activation and Akt inactivation, which together prevented proliferation of human osteosarcoma cells (MG63) and fibrosarcoma cells (HT1080). PTEN knockdown by siRNA reduced the effects of caffeine on Akt inactivation in osteosarcoma cells. These results indicate that the tumor suppressor PTEN signaling pathway contributes to the growth-inhibitory effect of caffeine on sarcoma cells. Our data suggest that caffeine and other drugs that act on this pathway could have promising therapeutic effects in the treatment of sarcoma patients. PMID- 21617856 TI - Glucosylceramide synthase, a factor in modulating drug resistance, is overexpressed in metastatic breast carcinoma. AB - Drug resistance causes treatment failure in approximately 50% of breast cancer patients with chemotherapy. Overexpression of glucosylceramide synthase (GCS) confers drug resistance in cancer cells, and suppression of GCS sensitizes cancers to chemotherapy in preclinical studies. Thus, GCS becomes a potential target to reverse drug resistance; however, little is known about GCS expression levels in normal tissues and whether GCS overexpression is associated with metastatic cancers. Herewith, we report our studies in GCS expression levels and breast cancer from patients. GCS levels were analyzed using cancer profiling arrays, breast cancer histo-arrays and quantitative RT-PCR in tumor tissues. We found that breast (18 exp. index) and other hormone-dependent organs (testis, cervix, ovary, prostate) displayed the lowest levels of GCS mRNA, whereas liver (52 exp. index) and other organs (kidney, bladder, stomach) displayed the highest levels of GCS. GCS mRNA levels were significantly elevated in tumors of breast, cervix, rectum and small intestine, as compared to each paired normal tissue. In mammary tissue, GCS overexpression was detected in breast cancers with metastasis, but not in benign fibroadenoma or primary tumors. GCS overexpression was coincident with HER2 expression (gamma2=0.84) in ER-negative breast adenocarcinoma. In tumor specimens, GCS mRNA was elevated by 4-fold and significantly associated with stage III (5/7), lymph node-positive (7/8) and estrogen receptor-positive breast cancers (7/9). GCS expression was significantly and selectively elevated in breast cancer, in particular in metastatic disease. GCS overexpression was highly associated with ER-positive and HER2-positive breast cancer with metastasis. Although a small study, these data suggest that GCS may be a prognostic indicator and potential target for the treatment of chemotherapy-refractory breast cancer. PMID- 21617857 TI - EGF stimulates cyclooxygenase-2 expression through the STAT5 signaling pathway in human lung adenocarcinoma A549 cells. AB - The epidermal growth factor receptor (EGFR) can be activated by several growth factors within the tumor microenvironment, and it can activate several signaling pathways. For tumor development, these EGFR-related signaling pathways may converge on several common nuclear transcription factors, one such transcription factor being STAT5. STAT5 plays an important role in the oncogenic signal transduction pathway in non-small cell lung cancer. In this study, we examined whether the epidermal growth factor (EGF) can stimulate cyclooxygenase-2 (COX-2) expression in human lung adeno-carcinoma A549 cells transfected with or without STAT5 siRNA or dominant-negative (DN)-STAT5, and identified the pathways involved in this response. We found that STAT5 siRNA significantly reduced EGF-induced COX 2 expression, and STAT5 phosphorylation. STAT5 phosphorylation predominantly mediates EGF-induced COX-2 promoter activity. STAT5 siRNA was found to inhibit COX-2 expression in resting A549 cells despite the absence of detectable activated phosphorylated STAT5. Using an adenoviral system, we expressed DN-STAT5 in human lung adenocarcinoma A549 cells in order to broaden the investigation and to determine the role of STAT5 in EGF-mediated COX-2 gene expression. The overexpression of DN-STAT5 significantly inhibited EGF-induced COX-2 expression, and we found that EGF induced the tyrosine phosphorylation of STAT5 and up- regulated COX-2 expression. DN-STAT5 also blocked COX-2 promoter activity. Our results demonstrate that EGF stimulates COX-2 expression in human lung adenocarcinoma A549 cells via the activation of the STAT5 pathway and that COX-2 expression may be independent of phosphorylated STAT5 in A549 cells in vitro. PMID- 21617858 TI - Growth response of human colorectal tumour cell lines to treatment with afatinib (BIBW2992), an irreversible erbB family blocker, and its association with expression of HER family members. AB - Despite the approval of the anti-epidermal growth factor receptor (EGFR) monoclonal antibodies (mAbs), cetuximab and panitumumab, for the treatment of colorectal cancer patients, there is currently no reliable predictive marker for response to therapy. In addition, the duration of response is often limited. Therefore, this study aimed to investigate the effect of afatinib, an irreversible erbB family blocker, as a single agent or in combination with cytotoxic drugs (5-fluorouracil, irinotecan and oxaliplatin) or mAb ICR62 on the proliferation of a panel of human colorectal tumour cell lines and the association between the expression levels of the EGFR family members and response to treatment. Of the cells examined, EGFR-overexpressing DiFi cells were the most sensitive to treatment with both afatinib (IC50=45 nM) and ICR62 (IC50=4.33 nM). Afatinib also inhibited the growth of other tumour cell lines with IC50 values which ranged from 0.33 uM (CCL-221) to 1.62 uM (HCT-116). A significant association was found between the co-expression of EGFR, human epidermal growth factor receptor (HER)-2 and HER-3 and response to treatment with afatinib (R=0.915, P=0.021). Treat-ment with afatinib and cytotoxic drugs was accompanied by an increase in the proportion of these cells in the sub-G0/G1 and in the S and G2/M phase of the cell cycle, respectively. We conclude that afatinib as monotherapy or in combination with other drugs shows activity in colorectal tumour cells and that determination of the co-expression of HER family members should be conducted in clinical trials using drugs targeting erbB signaling. This approach could lead to the identification of a specific subpopulation of cancer patients more likely to benefit from erbB-directed therapy. PMID- 21617859 TI - Lactate enhances motility of tumor cells and inhibits monocyte migration and cytokine release. AB - In solid malignant tumors, lactate has been identified as a prognostic parameter for metastasis and overall survival of patients. To investigate the effects of lactate on tumor cell migration, Boyden chamber assays were applied. We could show here that lactate enhances tumor cell motility of head and neck carcinoma cell lines significantly in a dose-dependent manner. The changes in tumor cell migration could be attributed to L-lactate or a conversion of lactate to pyruvate, as only these two substances were able to increase migration. Addition of D-lactate or changes in osmolarity or intracellular pH did not alter the migratory potential of the cells investigated. Because lactate was shown earlier to impair the penetration of dendritic cells in a tumor spheroid model, which is contrary to the response of the malignant cell population in the present study, we included blood monocytes in our assay as a highly motile immune cell type and precursor of tumor-associated macrophages. Interestingly, high levels of L lactate (20 mM) at a pH of 7.4 inhibited monocyte migration in the Boyden chamber system. In addition, cytokine release of TNF and IL-6 was inhibited. The obtained data suggest that high lactate content promotes tumor progression by contributing to the phenomenon of tumor immune escape and by enhancing the migratory potential of the malignant cell population which may directly be coupled to a higher incidence of metastasis. PMID- 21617860 TI - Clinicopathological and biological significance of CDC28 protein kinase regulatory subunit 2 overexpression in human gastric cancer. AB - CDC28 protein kinase regulatory subunit 2 (CKS2) is a cyclin-dependent kinase subunit (CKS) family member that participates in cell cycle regulation. Few studies have investigated its involvement in gastric cancer. In this study, we focused on the clinical and biological significance of CKS2 over-expression in gastric cancer. The expression of CKS2 mRNA was studied by real-time quantitative reverse transcription polymerase chain reaction, and the expression status in each tumor was examined to varify whether any correlation existed with clinical and pathological factors. In addition, an immuno-histochemical study was performed in selected samples. Moreover, we examined the impact of CKS2-siRNA in a gastric cancer cell line. A significantly higher expression of CKS2 mRNA was found in tumor tissues compared to paired normal tissues (p<0.01). Immunohistochemical analyses led to similar results. The overall five-year survival rate was significantly higher in the low CKS2 expression group (59.9%) than in the high CKS2 expression group (23.9%) (p<0.01). Univariate and multivariate analysis showed that CKS2 expression status was an independent prognostic factor (relative risk, 1.41; 95% confidence interval, 1.01-1.97; p<0.05). Moreover, the inhibition of cellular proliferation by CKS2-siRNA was observed in a gastric cancer cell line. CKS2 is one of the gastric cancer-related genes that correlates with biological aggressiveness and poor prognosis of gastric cancer. Thus, CKS2 is a possible candidate gene for diagnosis, as well as targeted molecular diagnosis and therapy in gastric cancer. PMID- 21617861 TI - Apoptotic death in curcumin-treated NPC-TW 076 human nasopharyngeal carcinoma cells is mediated through the ROS, mitochondrial depolarization and caspase-3 dependent signaling responses. AB - Curcumin, a potent candidate anticancer agent, is a dietary pigment (phenolic compound) derived from the food flavoring spice turmeric (Curcuma longa), and it has been shown to have inhibitory effects on tumor cells through anti proliferative and proapoptotic activities. However, there is no report showing curcumin-induced apoptotic cell death in human nasopharyngeal carcinoma cells in vitro. Thus, this study was performed to elucidate whether mitochondria and caspase cascades are involved in the modulation of apoptosis and cell cycle arrest in curcumin-treated NPC-TW 076 human nasopharyngeal carcinoma cells. The effects of curcumin on cell cycle arrest and apoptosis were measured by flow cytometry, and caspase-3 activity, apoptosis-associated protein levels and its regulated molecules were studied by flow cytometric assay and immunoblots. The results indicated that curcumin-induced G2/M phase arrest was associated with a marked decrease in the protein expression of cyclin A, cyclin B and cyclin dependent kinase 1 (Cdk1). Curcumin-induced apoptosis was accompanied with upregulation of the protein expression of Bax and downregulation of the protein levels of Bcl-2, resulting in dysfunction of mitochondria and subsequently led to cytochrome c release and sequential activation of caspase-9 and caspase-3 in NPC TW 076 cells in a time-dependent manner. These findings revealed that mitochondria, AIF caspase-3- dependent pathways play a vital role in curcumin induced G2/M phase arrest and apoptosis of NPC-TW 076 cells in vitro. PMID- 21617862 TI - Predicting the chemosensitivity of pancreatic cancer cells by quantifying the expression levels of genes associated with the metabolism of gemcitabine and 5 fluorouracil. AB - Gemcitabine (GEM) is the standard treatment for advanced/metastatic pancreatic cancer. However, there is a substantial subset of patients in whom the efficacy of GEM, when used as a single agent, is inadequate. Recently, the 5-fluorouracil (5-FU) prodrugs capecitabine and S-1 have been used as an alternative, either alone or in combination with GEM. The aim of the present study was to investigate the expression pattern of genes that render pancreatic cancer cells sensitive to GEM and 5-FU, and to identify markers for individualized chemotherapy, even in patients who have developed resistance. We investigated the correlation between the expression of genes associated with the metabolism of GEM and 5-FU, and sensitivity to these drugs in 15 human pancreatic cancer cell lines. We also established GEM- and 5-FU-resistant pancreatic cancer cell lines to investigate changes in the expression levels of these genes and the effects of one drug on cells resistant to the other. We found no correlation between pancreatic cancer cell sensitivity to either GEM- or 5-FU. GEM-resistant cells did not become resistant to 5-FU and vice versa. High expression of RRM1 (P=0.048) and TS x DPD (P=0.035) correlated significantly with sensitivity to GEM and 5-FU, respectively. 5-FU-resistant cells expressed significantly higher levels of TP than parental cells (P<0.05). In conclusion, pancreatic cancer cells showed no cross-resistance to GEM and 5-FU. Quantitative analyses of RRM1, TP, DPD and TS mRNA levels in pancreatic cancer cells may be useful for predicting their sensitivity to GEM and 5-FU. PMID- 21617863 TI - Anti-aging effects of extracts prepared from salmon nasal cartilage in hairless mice. AB - The skin anti-aging effects of orally administered salmon nasal cartilage extract (SNCE), which includes abundant proteoglycan, were investigated using a hairless mouse skin-aging model, in which aging was caused by repetitive ultraviolet B (UV B) irradiation. Physiological analysis of the skin surface following repetitive UV-B irradiation of 8 weeks revealed inhibition of erythema levels and reduction of transepidermal water loss (TEWL) due to oral administration of SNCE. Similarly, inhibitory actions of epidermal and dermal hypertrophy were revealed by hematoxylin and eosin staining. Furthermore, effects on the hydration level of the skin surface by SNCE administration were indicated at 4 weeks of UV-B irradiation, but greater effects were not apparent. These results indicate that SNCE may serve as an anti-aging agent for healthy skin. PMID- 21617864 TI - Histone deacetylase inhibitors induce thymidine phosphorylase expression in cultured breast cancer cell lines. AB - Thymidine phosphorylase (TP) is an enzyme involved in thymidine synthesis and degradation. The expression of this enzyme has been proposed as a predictive factor for the therapeutic benefit of capecitabine, which is a precursor of the drug 5'-fluorouracil. In fact, TP is the rate-limiting enzyme in the activation of capecitabine. Therefore, higher levels of TP are expected to sensitize cancer cells to capecitabine treatment. In the present study, using breast cancer cell lines, we found a correlation between TP mRNA and protein levels, suggesting that compounds able to increase TP gene expression also increase protein levels. Accordingly, we demonstrated that treatment of breast cancer MCF7 and MDA231 cell lines with histone deacetylase inhibitors, tricostatin A and suberoylanilide hydroxamic acid, increased TP both at the mRNA and protein level. The effects of histone deacetylase inhibitors were not found to occur via the cytokine TNFalpha, a known inducer of TP expression. Our findings suggest a strategy to sensitize breast cancer cells to capecitabine treatment. PMID- 21617865 TI - Sulforaphane inhibits the growth of KPL-1 human breast cancer cells in vitro and suppresses the growth and metastasis of orthotopically transplanted KPL-1 cells in female athymic mice. AB - The anticancer effects of sulforaphane (SFN), which is found in cruciferous vegetables, were studied on KPL-1 human breast cancer cells in vitro and in vivo. Cell proliferation in vitro was assessed by a 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay, and tumor growth and metastasis in vivo were examined in orthotopically (right thoracic mammary fat pad) transplanted KPL 1 cells in female athymic BALB/c mice. The MTT assay showed that SFN directly inhibited KPL-1 cell growth in vitro (IC50 at 48 h, 19.1 uM; IC50 at 72 h, 17.8 uM). Athymic mice received a KPL-1 cell transplant, and SFN treatment (intraperitoneal injection of 25 or 50 mg/kg SFN) was started the next day. Mice received five injections each week during the 26-day experimental period (for a total of 20 injections). Compared with the SFN-untreated controls, SFN suppressed primary tumor growth. At the termination of the experiment, the final tumor volume was 686+/-94 mm3 for the control group, 516+/-70 mm3 (75% of control value) for the 25 mg/kg SFN group and 351+/-55 mm3 (51% of control value) for the 50 mg/kg SFN group. The final tumor weight was 571+/-69 mg in the control group, 416+/-63 mg (73% of the control value) in the 25 mg/kg SFN group and 338+/-56 mg (59% of the control value) in the 50 mg/kg SFN group. SFN caused a dose-dependent decrease in the proliferation ratio and an increase in the apoptotic ratio of the primary tumor cells. SFN treatment tended to reduce regional (axillary) lymph node metastasis. Treatment with 50 mg/kg SFN significantly inhibited KPL-1 cell growth in vivo by suppressing cell proliferation and inducing apoptosis, and it tended to reduce axillary lymph node metastasis of KPL-1 human breast cancer cell xenografts in female athymic mice. PMID- 21617866 TI - Expression of histone deacetylase 1 and metastasis-associated protein 1 as prognostic factors in colon cancer. AB - Histone deacetylase 1 (HDAC1) and metastasis-associated protein 1 (MTA1) form the nucleosome remodeling and histone deacetylation (NuRD) complex and may possibly play a central role in cancer development. However, limited data has been reported regarding the expression of both HDAC1 and MTA1. The aim of the present study was to clarify the clinical role of HDAC1 and MTA1 expression in colon cancer. Seventy-four patients with colon cancer, who underwent colectomy at our institution, were enrolled in this study. Expression of HDAC1 and MTA1 was examined immunohistochemically. The patients were divided into four groups: HDAC1 positive group (n=58), HDAC1-negative group (n=16), MTA1-positive group (n=38) and MTA1-negative group (n=36). Clinicopathological factors and survival rates were compared between the groups. Regarding the clinicopathological factors, the depth of tumor invasion and stage correlated significantly with HDAC1 expression (p<0.05). Age, depth of tumor invasion and vascular invasion tended to correlate with MTA1 expression. The 5-year survival rate in the HDAC1-positive group (55.1%) was significantly worse compared to the HDAC1-negative group (86.5%) (p<0.05), and the 5-year survival rate of the MTA1-positive group (50.5%) was significantly worse than that of the MTA1-negative group (73.1%) (p=0.05). In patients with stages II-IV and curability A, B, the survival rate in those with HDAC1-positive expression was significantly worse than those with HDAC1-negative expression (p<0.05), and the survival rate of the MTA1-positive group tended to be worse than that of the MTA1-negative group (p=0.07). Overall survival in both the HDAC1 and MTA1-positive groups was significantly worse than overall survival of the other groups (p<0.05). Disease-free survival in both the HDAC1- and MTA1 positive groups, among patients with stages II-IV and curability A, B, was also significantly worse than that of the other groups (p<0.05). HDAC1 and MTA1 expression levels were significantly related to poorer prognosis. Therefore, HDAC1 and MTA1 expression levels are potential prognostic indicators for colon cancer. PMID- 21617867 TI - Inhibition of nuclear factor-kappaB activity by small interfering RNA in esophageal squamous cell carcinoma cell lines. AB - Chemotherapy with 5-fluorouracil (5-FU) is commonly used in combination therapy for esophageal squamous cell carcinoma (ESCC), but its efficacy is limited in certain patients. Recent studies suggest that constitutive activation of nuclear factor-kappaB (NF-kappaB) has a critical role in tumorigenesis and is associated with poor prognosis and resistance to chemoradiation therapy in many types of human cancers. In the present study, we evaluated the effect of small interfering RNA targeting NF-kappaB (NF-kappaB siRNA) combined with 5-FU on the proliferation of two cell lines of cultured ESSCs. Immunofluorescence and immunoblot analyses revealed that the NF-kappaB protein was localized mostly in the cytoplasm of ESCCs. When cultured ESCCs were exposed to tumor necrosis factor-alpha, NF-kappaB was transferred to the nucleus and activated. ESCCs with activated NF-kappaB had poor sensitivity to 5-FU. When cells were transfected with NF-kappaB siRNA, the levels of NF-kappaB protein were significantly decreased in the cytoplasm and the nucleus. Transcriptional activity of NF-kappaB was significantly suppressed in cells treated with 5-FU and NF-kappaB siRNA compared to cells treated with 5-FU alone. 5-FU consistently suppressed proliferation of ESCCs in a dose-dependent manner, and this effect was significantly enhanced when combined with NF-kappaB siRNA. These results suggest that combination therapy of 5-FU with NF-kappaB siRNA may provide a new therapeutic option for ESCC. PMID- 21617868 TI - Tissue factor is involved in retinoblastoma cell proliferation via both the Akt and extracellular signal-regulated kinase pathways. AB - Tissue factor (TF) is known to play a role in tumor progression. In retinoblastoma, the expression and role of TF has not been determined yet. Herein, we demonstrated for the first time that TF is closely related to the proliferation of retinoblastoma cells, which could be therefore effectively suppressed by blockade of the TF pathway. TF was selectively expressed on the areas of highly mitogenic activity in an orthotopic transplantation mouse model of retinoblastoma. In addition, the levels of TF expression in retinoblastoma cells were elevated after FGF2 treatment, whereas the proliferative effect of FGF2 on retinoblastoma cells was significantly inhibited by blockade of the TF pathway via TF pathway inhibitor (TFPI). Interestingly, retinoblastoma cells cultured with FGF2 showed increased phosphorylation of both Akt and ERK1/2. Addition of TFPI nearly abolished the FGF2-induced phosphorylation of Akt and ERK1/2 in retinoblastoma cells. Therefore, our data suggest that TF expression in retinoblastoma cells is closely related to tumor cell proliferation and TFPI has the potential to inhibit retinoblastoma cell proliferation via the inhibition of both Akt and ERK1/2 activation. PMID- 21617869 TI - Expression of KL-6/MUC1 in pancreatic cancer tissues and its potential involvement in tumor metastasis. AB - Aberrant expression of KL-6/MUC1 mucin has been proven to be associated with poorer tumor behavior in many carcinomas. The aim of this study was to evaluate the expression of KL-6/MUC1 in pancreatic cancer tissues and its potential involvement in tumor metastasis. The expression of KL-6/MUC1 in 18 cases of pancreatic ductal carcinoma (PDC), 5 cases of intraductal papillary mucinous tumor (IPMT), and 3 cases of islet cell tumor was detected by immunohistochemical staining. To determine the impact of loss of KL-6/MUC1 expression on pancreatic tumor progression, an siRNA targeting MUC1 was synthesized and transfected into Panc-1 cells and Capan-1 cells to knock down KL-6/MUC1 expression. Down regulation of KL-6/MUC1 expression was detected by reverse transcription polymerase chain reaction (RT-PCR) and western blotting. E-cadherin and KL-6 mucin co-expression was detected by immunofluorescence. The expression of E cadherin and E-cadherin/beta-catenin comlex was determined by immunoprecipitation. Cell invasive abilities were detected by invasion assay. Positive KL-6/MUC1 staining was observed in all 18 PDC cases (18/18,100.0%) and 1 metastatic IPMT case (1/5, 20.0%). The results suggested that KL-6/MUC1 overexpression may be associated with more aggressive tumor behavior, although the cases of pancreatic cancer tissues in this study are limited. RT-PCR and western blotting showed that both KL-6/MUC1 mRNA and protein can be effectively silenced. Following KL-6/MUC1 knockdown, E-cadherin expression increased. Also, E cadherin/beta-catenin complex expression increased and the invasive ability of the cells decreased. This study indicated that overexpression of KL-6/MUC1 in pancreatic cancer tissues may be associated with metastasis of pancreatic cancer by regulating E-cadherin and E-cadherin/beta-catenin complex expression. PMID- 21617870 TI - Effects of 5,6-benzoflavone, indole-3-carbinol (I3C) and diindolylmethane (DIM) on chemically-induced mammary carcinogenesis: is DIM a substitute for I3C? AB - The abilities of 5,6-benzoflavone (5,6-BF, a synthetic flavonoid), indole-3 carbinol (I3C, a plant derived product) or diindolylmethane (DIM, a condensation product of I3C) to alter the induction of mammary cancers induced by the carcinogens 7,12-dimethylbenzanthracene (DMBA) or N-methyl-N-nitrosourea (MNU) were evaluated. Interestingly, the first two agents act as aryl hydrocarbon receptor (AhR) agonists, while DIM does not. The agents were initially examined for their ability to inhibit DMBA-induced mammary carcinogenesis. Agents were administered for 14 days starting 7 days prior to a single dose of the carcinogen. Evaluated over an extensive range of doses (165, 550 and 1650 ppm in the diet), 5,6-BF caused a dose-dependent decrease of mammary cancers. In addition, 5,6-BF at doses of 1650 and 165 ppm in the diet blocked the induction of DMBA-induced DNA adducts in the mammary gland by approximately 85% and 45%, respectively. In contrast, DIM (180 or 20 mg/kg BW/day) failed to block induction of DMBA tumors. The effect of these agents on the promotion/progression phase of carcinogenesis using the MNU mammary cancer model was also determined. 5,6-BF (1650 or 165 ppm in the diet), I3C (180 or 60 mg/kg BW/day administered by gavage), or DIM (180 or 60 mg/kg BW/day by gavage) were initiated 5 days after the administration of MNU, and continually thereafter. 5,6-BF decreased MNU- induced mammary tumor multiplicity by 40-60%. I3C reduced tumor multiplicity at the high dose, while DIM at either dose had minimal effects on tumor multiplicity. Thus, 5,6-BF and I3C were highly effective against initiation of DMBA-induced mammary carcinogenesis, and were also effective against MNU-induced tumors during the promotion/progression phase of carcinogenesis. In contrast, DIM had minimal effects in either model; arguing that administration of DIM is not analogous to administration of I3C. PMID- 21617871 TI - Effect of temozolomide on cell viability in gonadotroph adenoma cell lines. AB - Invasive pituitary adenomas are usually refractory to routine neurosurgery, radiosurgery or medications, and alternative therapies are needed. The effects of temozolomide (TMZ) on the inhibition of gonadotroph adenoma cell viability and hormone secretion were evaluated. Cell viability and IC50 values were evaluated after alphaT3-1 cells were treated with TMZ (31.25-1000 uM) or vehicle for 0-72 h. Cell cycle changes and the extent of apoptosis were detected using flow cytometry, TUNEL and TEM. The molecular mechanism of TMZ action was investigated by the Caspase-Glo(r) assay and immunoblotting. Gonadotropin secretion was assessed using an immunoassay system. TMZ dose- and time-dependently suppressed cell proliferation (P<0.01 vs. control, 250 uM, 24 h) and induced S-phase accumulation and G2/M-phase arrest (P<0.05 vs. control, 250 uM, 24 h). Early apoptotic cells increased following a 24-h TMZ incubation (P<0.001 vs. control, 250 uM), consistent with TEM and TUNEL detection that exhibited morphological features of apoptosis. TMZ (250 uM) increased the level of caspase-3/7 by 6-fold, caspase-9 by 7-fold and caspase-8 by 3-fold after a 24-h incubation, while it attenuated Bcl-2 expression (P<0.001 vs. control) and raised the proteolysis of PARP. Both FSH and LH levels were significantly decreased by TMZ (P<0.01 vs. control, 250 uM, 24 h). TMZ inhibited cell proliferation and hormone secretion, and induced cell cycle arrest and apoptotic cell death in gonadotroph adenoma cells via both death receptor and mitochondrial pathways, suggesting that it may represent a useful medical management strategy of invasive gonadotroph adenomas. PMID- 21617872 TI - Pigment epithelium-derived factor inhibits angiogenesis and growth of gastric carcinoma by down-regulation of VEGF. AB - Previous studies have shown that pigment epithelium-derived factor (PEDF) is an antitumor candidate with anti-oxidative, anti-inflammatory and anti-angiogenesis properties. However, the effect of PEDF on gastric carcinoma has not been elucidated. MTT assay and Annexin V/PI staining were performed. Immunohistochemistry was applied to detect microvessel density (MVD) of a xenograft model. The protein levels of vascular endothelial growth factor (VEGF) were examined by western blot analysis and hypoxia-inducible factor-1alpha (HIF 1alpha) translocation was investigated by immunofluorescence. Results showed that growth and angiogenesis of gastric carcinoma were suppressed after PEDF injection. PEDF could not directly suppress proliferation or induce apoptosis of gastric carcinoma cells. However, the expression of VEGF both in tumor tissues and gastric carcinoma cells was down-regulated by PEDF. The amount and nuclear translocation of HIF-1alpha, the transcription factor of VEGF, was also inhibited by PEDF. In conclusion, PEDF suppresses angiogenesis and growth of gastric carcinoma by down-regulating HIF-1alpha and VEGF and may have potential for gastric carcinoma treatment. PMID- 21617873 TI - Radiotherapy and wound healing: principles, management and prospects (review). AB - Radiation therapy is a major therapeutic modality in the management of cancer patients. Over 60% of these patients receive radiotherapy at some point during their course of treatment and over 90% will develop skin reactions after therapy. Problematic wound healing in radiation-damaged tissue constitutes a major surgical difficulty and despite all efforts, irradiated skin remains a therapeutic challenge. This review provides an overview of the fundamental principles of radiation therapy with regards to the wound healing in normal and irradiated skin. Furthermore, it presents techniques that describe how to prevent and manage skin side effects as well as prospects that may improve cutaneous wound repair in general and in irradiated skin. PMID- 21617874 TI - Activity of aspirin analogues and vanillin in a human colorectal cancer cell line. AB - Colorectal cancer is the third most common cancer and is associated with significant morbidity and mortality. Epidemiological and animal studies indicate that regular acetylsalicylic acid (aspirin) intake is associated with a reduction in the incidence of colorectal cancer. Acetylsalicylic acid (ASA) has also been shown to inhibit colorectal cancer cell proliferation in vitro. The molecular basis for this specific cytotoxicity is an area of considerable debate. To investigate the toxicity of salicylates, the sensitivity of the DNA mismatch repair proficient SW480 human colorectal cancer cell line to four categories of compounds with varying degrees of structural similarity to acetylsalicylic acid was tested. These compounds were: i) salicylic acid analogues with substituents at the 5-position; ii) ASA analogues with extended chain lengths in the acyl group; iii) vanillin (4-hydroxy-3-methoxybenzaldehyde; and iv) bis(2 carboxyphenyl) succinate (BCS) and structurally similar derivatives thereof. It was found that compounds with amino and acetamido substituents at the salicylate 5-position were less toxic than ASA itself. Modifications to the length of the hydrocarbon chain in the acyl groups of ASA analogues also marginally reduced toxicity. Vanillin exhibited relatively limited toxicity against the SW480 colorectal cancer cell line. Commercially available and in-house synthesised BCS (diaspirin) were notably more inhibitory to cell growth than ASA itself, yet retained substantial specificity against colorectal cancer cell lines vs. non colorectal cancer cell lines. BCS and ASA were toxic to SW480 cells through initiation of necrotic and apoptotic pathways. Fumaroyldiaspirin and benzoylaspirin exhibited greater toxicity than ASA against the SW480 cell line. A novel method for synthesis of BCS, a compound that has erratic commercial availability, is described. We propose that the anti-inflammatory and anticancer capacity of BCS and the other analogues described herein is worthy of investigation. PMID- 21617875 TI - Fenofibrate attenuates contact-stimulated cell motility and gap junctional coupling in DU-145 human prostate cancer cell populations. AB - In the present study, we investigated the effects of fenofibrate on the invasive potential of DU-145 human prostate cancer cells in the context of gap junctional intercellular coupling and the formation of reactive oxygen species. Time-lapse analyses of cell motility, accompanied by tests of cell viability, membrane microviscosity, reactive oxygen species accumulation and the function of gap junctional protein connexin 43 were performed in monolayer cultures of DU-145 cells following fenofibrate administration. Fenofibrate inhibited the motility of DU-145 cells and attenuated gap junctional intercellular coupling in a manner independent of its effects on cell viability, PPARalpha activation and cell membrane micro-viscosity. Instead, N-acetyl-L-cysteine, a scavenger of reactive oxygen species, restored cell motility and gap junctional coupling in fenofibrate treated DU-145 cell populations. These data indicate that two parameters crucial for cancer cell metastatic potential, i.e. cell motility and gap junctional coupling, are inhibited by fenofibrate. Thus, fenofibrate affects prostate cancer cell invasion via an orchestrated action on versatile cancer cell properties determining this process. A novel mechanism of anti-invasive activity of fenofibrate, which depends on its interference with cell motility and the function of gap junctions regulated by reactive oxygen species, is suggested. PMID- 21617876 TI - CD44 as a stem cell marker in head and neck squamous cell carcinoma. AB - In the recent past, evidence is increasing indicating the existence of a subpopulation of resistant tumor cells in head and neck squamous cell carcinoma (HNSCC) that cannot be eradicated by established antineoplastic treatments. These cancer stem cells (CSCs) have features of somatic stem cells such as selfrenewal, proliferation and differentiation. CD44+ cells in tumors of the head and neck are referred to as CSCs of HNSCC. Expression profiling of CD44 in 29 HNSCC tumors was performed by fluorescence microscopy. ELISA analysis was performed to detect concentration of soluble CD44 in the peripheral blood of 29 HNSCC patients and 11 healthy controls. Expression of CD44 was determined in all HNSCC tissue samples (n=29). In all samples a surface staining pattern was found. The concentration of CD44 in the peripheral blood of HNSCC patients was significantly higher compared to a healthy control group (mHNSCC =13.5 +/- 0.5 ng/ ml; mCont = 9.3 +/- 0.6 ng/ml; P=0.6 x 10(-12)). The role of CD44 as a marker for CSCs in HNSCC remains to be ascertained. Further experiments might reveal its role as a diagnostic and prognostic factor, and possibly as a therapeutic target. PMID- 21617877 TI - beta-catenin plays a key role in metastasis of human hepatocellular carcinoma. AB - Currently, there are no diagnostic or metastatic markers that can be used in early diagnosis and treatment of human hepatocellular carcinoma (HCC). The aim of this study was to find a molecular marker that regulated migration and metastasis in HCC. We analyzed the gene expression of beta-catenin, c-Myc and IL-8 in human HCC tissue by RT-PCR and immunohistochemistry and analyzed five variously differentiated HCC cell lines by Western blotting and migration and invasion assays to find markers for HCC diagnosis and HCC metastasis. mRNA expression of beta-catenin was significantly higher in the tumor area compared to the non-tumor area and was more abundant in specimens of late-stage HCC. Immunohistochemistry revealed that the translocation of beta-catenin into the nucleus was closely correlated with IL-8 protein levels and tumor stage. Similarly, the level of expression and nuclear translocation of beta-catenin was greater in HA22T cells with high proliferative activity than in HCC cell lines with low proliferative activity (PLC, Hep3B, HepG2). Knockdown of the beta-catenin gene with beta catenin antisense oligonucleotides resulted in inhibition of cell migration and invasion of HA22T cells. Taken together, these results suggest that beta-catenin may be a suitable diagnostic marker of metastasis in human HCC. PMID- 21617878 TI - Human FAT1 cadherin controls cell migration and invasion of oral squamous cell carcinoma through the localization of beta-catenin. AB - FAT1 [Homo sapiens FAT tumor suppressor homolog 1 (Drosophila)] is an intrinsic membrane protein classified as a member of the cadherin superfamily. The FAT1 gene is a tumor suppressor in humans as well as being the pivotal gene for cell morphogenesis and migration. Deletion of this gene could play a role in the characteristics of oral squamous cell carcinomas (OSCCs), involving cell adhesion, migration and/or invasion. This study investigated the mechanisms by which FAT1 is involved in the biological behavior of OSCCs. First, a rat monoclonal antibody was developed against a FAT1 intra-cellular domain epitope, and used for an immunohistochemical study of FAT1 in clinically obtained OSCC samples. FAT1 was localized at lamellipodial edges or cell-cell boundaries in normal cells and well differentiated OSCCs, but showed a diffuse cytoplasmic and nuclear distribution in moderately-poorly differentiated OSCCs. FAT1-siRNA was transfected into OSCCs resulting in a drastic inhibition of cell migration and invasion based on the suppression of FAT1 expression and disorganized localization of beta-catenin which is associated with cell polarity and migration. These results suggested that FAT1 may be involved in the migration and invasion mechanisms of OSCCs and, therefore, it could be an important target for the development of new therapeutic strategies. PMID- 21617879 TI - Prognostic implications of the DNA damage response pathway in glioblastoma. AB - Genomic instability and resistance to genotoxic therapies for glioblastoma (GBM) suggest aberrant DNA damage response (DDR), since DDR maintains the genomic integrity against genotoxic insults including anti-tumor therapies. To elucidate the biological and clinical meaning of DDR in GBM, we retrospectively investigated the immunohistochemical expression of DDR proteins (ATM, Chk1, Chk2, TopBP1, Rad17, p53, Nbs1, MDC1, gammaH2AX and RPA1) in 69 GBM surgical samples and their relation with GBM patient survival. Remarkably, higher expression of ATM revealed significantly longer overall survival (OS) and progression-free survival (PFS) (p<0.05). Upon multivariate analysis, expression level of ATM was an independent factor for longer OS (p=0.020) and longer PFS (p=0.019). Since ATM induces cell cycle arrest or apoptosis through cell cycle regulators in response to genotoxic insults, these results indicate that aberrant DDR signaling through ATM in GBM may be associated with resistance to genotoxic anti-tumor therapeutics. Conclusively, we emphasize that the identification of DDR machinery, which can be involved in unstable genomic status or genotoxic therapies in GBM, is very important to predict patient outcome. PMID- 21617880 TI - Extraskeletal Ewing's sarcoma of the thoracic epidural space: case report and review of the literature. AB - The occurrence of primary extraskeletal Ewing's sarcoma (EES) of the spinal epidural space has been rarely reported in the literature. The clinical, radiologic and pathologic features of a case of EES occurring in the thoracic epidural space are presented. A 37-year-old woman presented with a one-year history of back pain. Magnetic resonance imaging demonstrated an epidural mass at the T8-9 level. Laminectomy and partial resection of the tumor were performed. The differential diagnosis of a spinal epidural mass is broad. Histopathological and molecular cytogenetic examinations confirmed an EES arising from the thoracic epidural space. Despite receiving both chemotherapy and radiotherapy, the patient died of respiratory insufficiency due to medulla oblongata metastasis 22 months after the initial consultation. Awareness of this entity will allow this rare diagnosis to be considered and facilitate appropriate management. A review of the literature on spinal epidural EES is also presented. PMID- 21617881 TI - Loss of 3p26.3 is an independent prognostic factor in patients with oral squamous cell carcinoma. AB - Oral squamous cell carcinoma (OSCC) is a common malignancy worldwide and the prognosis for patients with advanced-stage OSCC is particularly poor. To identify DNA copy number aberrations and candidate genes associated with a poor or favorable outcome, we analyzed the genome profiles of OSCC tumors by array-based comparative genomic hybrid-ization (A-CGH). This technique uses DNA microarray technology to detect genomic copy number variations at a higher resolution level than chromosome-based CGH. Fifty patients with primary OSCCs were included in the study. Of these 50 patients, 37 were treated surgically and 13 were treated without surgery and had received irradiation and/or chemotherapy. All samples were analyzed by A-CGH. Gains were detected frequently (>50%) at chromosomal regions 5p15.33, 7p22.3, 8q21.1-24.3, 9q34.3, 11q13, 16p13.3 and 20q13.3. Losses were frequently detected at 3p22, 3p14 and 4q35.2. High-level gains were recurrently (>10%) detected at each of 5p15, 7p22, 7p11, 8q24, 11q13, 11q22 and 22q11. Gains of 2p25.1, 11p15, 16p13.3, 16q24.3 and 20q13.3 were inversely correlated with nodal metastasis. In 37 of the 50 OSCC patients treated with surgery, gains of 8q12.1-24.22 and losses of 3p26.2-3 were associated with disease-specific survival (p<0.01). Loss of a 0.2 Mb chromosomal region in 3p26.3 was associated with a poor prognostic outcome in the Kaplan-Meier analysis (p<0.01 by the log-rank test). Multivariate analysis revealed that loss of 3p26.3 is an independent prognostic factor (p<0.01) of OSCC. Loss of a 0.2 Mb chromosomal region in 3p26.3 including the CHL1 (cell adhesion molecule with homology to L1CAM1) gene was identified as a novel potential marker for predicting the prognosis of patients with OSCC. PMID- 21617882 TI - Quercetin induces apoptosis in the methotrexate-resistant osteosarcoma cell line U2-OS/MTX300 via mitochondrial dysfunction and dephosphorylation of Akt. AB - Quercetin is the most abundant polyphenolic flavonoid found in plants. Several studies suggest that it has potent anticancer effects. The present study examines the apoptosis-inducing activity and the underlying mechanism of action of quercetin in a methotrexate (MTX)-resistant osteosarcoma model. Our results showed that quercetin inhibited cell viability in a dose-dependent manner and there was no cross-resistance between MTX and quercetin in U2-OS/MTX300 cells. The induction of apoptosis was observed by flow cyto-metry and fluorescence staining experiments. Quercetin-induced apoptosis was accompanied by a significant reduction of mitochondrial membrane potential, release of mitochondrial cytochrome c to the cytosol, activation of caspase-3, down regulation of Bcl-2, p-Bad and up-regulation of Bax. A remarkable dephospho rylation of Akt was also detected after quercetin treatment. Furthermore, transduction with constitutively active Akt protected against the quercetin induced dephosphorylation of Akt and Bad as well as poly(ADP-ribose)polymerase (PARP) degradation, while combined treatment with quercetin and LY294002 enhanced the dephosphorylation of Akt, Bad and PARP cleavage in U2-OS/MTX300 cells. Taken together, our results demonstrate that quercetin-induced apoptosis in the MTX resistant osteosarcoma cells U2-OS/MTX300 was mediated via mitochondrial dysfunction and dephosphorylation of Akt. PMID- 21617883 TI - Fasting glucose cutoff point: where does the risk terminate? Tehran lipid and glucose study. AB - The aim of this study is to determine safe fasting plasma glucose (FPG) levels. We included data on 5,960 individuals aged >=20 years at baseline with at least one follow-up examination. Diabetes was ascertained in accordance with American Diabetes Association criteria, using standard 2-h post-challenge plasma glucose test. Multivariate restricted cubic splines Weibull regression was implemented for interval-censored survival data on incident diabetes. We used Harrell's C statistic for discrimination, Nam-D'Agostino chi(2) for calibration, and Royston's R(2) for variations in the outcome explained by models. During a 6-year median follow-up, 369 incident cases of diabetes were ascertained. Family history of diabetes, systolic blood pressure, waist-to-height ratio, and triglyceride-to high-density lipoprotein cholesterol ratio, independent of FPG and each other remained associated with incident diabetes. The cubic splines model achieved good calibration (chi(2) = 12.1) and discrimination (C = 0.828) and explained 75% of variation in the time until incident diabetes. A J-shaped FPG-diabetes relationship was observed. Descending arm of the dose-response relationship curve corresponded to increasing FPG levels up to 4.0 mmol l(-1), where it started increasing. The risk of incident diabetes decreased with decreasing levels of FPG down to 4.0 mmol l(-1), where the risk stopped decreasing. Multivariate-adjusted risk of incident diabetes was zero at FPG = 5.05 mmol l(-1). Although currently there is no firm evidence to show that early intervention among individuals with the elevated FPG levels could prevent or delay onset of diabetes, individuals with FPG levels below 5.05 mmol l(-1) could be safely reassured about their near term risk of developing incident diabetes and screened on a less frequent basis. PMID- 21617884 TI - Does 20-min arm crank ergometer exercise increase plasma interleukin-6 in individuals with cervical spinal cord injury? AB - Interleukin-6 (IL-6) is produced by contracting skeletal muscles and then released into the circulation and considered to mediate the health benefits of exercise against chronic diseases. Individuals with spinal cord injury (SCI) are reported to be at higher risk of developing metabolic diseases. We investigated the IL-6 responses to 20-min arm crank ergometer exercise at 60% of maximum oxygen consumption in eight trained individuals with cervical SCI (CSCI) between C6 and C7, and eight able-bodied trained healthy subjects. The plasma concentrations of IL-6, adrenaline, prostaglandin E(2) and cortisol were measured before, immediately after the exercise, 1 and 2 h after exercise. At rest, the plasma adrenaline concentration was significantly lower in individuals with CSCI than in able-bodied subjects (P < 0.01). On the other hand, the concentration of IL-6 was significantly higher at rest in individuals with CSCI (2.18 +/- 0.44 pg/ml, mean +/- SEM) than the control (1.02 +/- 0.22 pg/ml, P < 0.05). In able bodied subjects, the plasma adrenaline concentration increased significantly immediately after the exercise (P < 0.01) and returned to the baseline level at 1 h after exercise, and the plasma IL-6 level increased significantly at 1 h after exercise (1.91 +/- 0.28 pg/ml, P < 0.05) and returned to the baseline level at 2 h after exercise. In contrast, adrenaline and IL-6 levels were steady throughout the study in individuals with CSCI. The lack of exercise-related IL-6 response in individuals with CSCI could be due to muscle atrophy and sympathetic nervous system dysfunction. PMID- 21617885 TI - The Woven EndoBridge cerebral aneurysm embolization device (WEB II): initial clinical experience. AB - INTRODUCTION: The Woven Endobridge (WEB II) device (Sequent Medical, Inc., Aliso Viejo, CA, USA) is an intra-saccular, oblate, braided-wire embolization device designed to provide flow disruption at the aneurysm neck-parent artery interface. The purpose of this study was to evaluate the acute and short-term performance of the WEB II device regarding the immediacy, degree, and durability of aneurysm occlusion in two patients. METHODS: The WEB II device was implanted in one patient with an unruptured MCA trifurcation aneurysm and one patient with an unruptured basilar tip aneurysm. The degree of intra-aneurysmal flow disruption was graded based on serial digital subtraction aneurysm angiography performed over 30 min immediately following device implantation and at 8 weeks. Immediate and 8-week post-treatment CT and 3-T MRI studies were also performed. RESULTS: Delivery and deployment of the WEB II device was technically straightforward and achieved without complications. Neither device required retrieval or repositioning after full deployment. There were no peri-procedural thrombembolic or hemorrhagic complications. In both cases, complete aneurysm occlusion was observed within minutes of device deployment. Short-term angiographic follow-up confirmed stable complete occlusion at 8 weeks. CONCLUSION: Early technical and clinical results from the first WEB II cases have been encouraging and suggest that the intra-saccular deployment of self-expanding, compliant, cylindrical, high-density, braided metallic mesh constructs may represent a feasible approach for the endovascular treatment of cerebral aneurysms. PMID- 21617886 TI - The Third Consensus Conference on the treatment of aplastic anemia. PMID- 21617887 TI - Single versus multiple dose intravenous immunoglobulin in combination with LED phototherapy in the treatment of ABO hemolytic disease in neonates. AB - Intravenous immunoglobulin (IVIG) has been found to decrease hemolysis in neonatal jaundice due to blood group incompatibility, but a consensus on its usage has not been reached. We conducted a study to compare single versus multiple dose of IVIG in combination with light emitting diode (LED) phototherapy in patients with neonatal jaundice secondary to ABO blood incompatibility, and compared the efficacy of these treatments with that in a group of patients who received LED phototherapy solely. Thirty-nine term neonates with ABO blood group incompatibility were enrolled in the study. Group I received one dose of IVIG (1 g/kg) and LED phototherapy, and group II two doses of IVIG (1 g/kg) and LED phototherapy, whereas group III received LED phototherapy only. In group I, exchange transfusion was performed in one patient (6%) and in group II in one patient (10%). In the control group, none of the patients required exchange transfusion. Duration of LED phototherapy was 4.3 +/- 0.7 days in group I + II (IVIG group), 3.9 +/- 0.6 days in group III (P = 0.06). Lowest hematocrit level in group I + II was 35.0 +/- 7.8 and group III was 38.9 +/- 4.2, this was statistically significant (P = 0.034). IVIG therapy, single or multiple, did not affect exchange transfusion, need of erythrocyte transfusion and hospitalization time when used in combination with LED phototherapy in the treatment of ABO hemolytic jaundice in neonates. PMID- 21617888 TI - Recombinant AAV2-mediated beta-globin expression in human fetal hematopoietic cells from the aborted fetuses with beta-thalassemia major. AB - Genetic correction of autologous hematopoietic stem cells has been proposed as an attractive treatment method for beta-thalassemia. Our previous study has shown that recombinant adeno-associated virus 2 (rAAV2) efficiently transduces human fetal liver hematopoietic cells, and mediates the expression of the human beta globin gene in vivo. In this study, we investigated whether rAAV2 could also mediate the expression of normal beta-globin gene in human hematopoietic cells from beta-thalassemia patients. Human hematopoietic cells were isolated from aborted beta-thalassemia major fetuses, transduced with rAAV2-beta-globin, and then transplanted into nude mice. We found that rAAV2-beta-globin transduced human fetal hematopoietic cells, as determined by allele-specific PCR analysis. Furthermore, beta-globin transgene expression was detected in human hematopoietic cells up to 70 days post-transplantation in the recipient mice. High-pressure liquid chromatography analysis showed that human beta-globin expression levels increased significantly compared with control, as indicated by a 1.2-2.8-fold increase in the ratio of beta/alpha-globin chain. These novel data demonstrate that rAAV2 can transduce and mediate the normal beta-globin gene expression in fetal hematopoietic cells from beta-thalassemia patients. Our findings further support the potential use of rAAV-based gene therapy in the treatment of human beta-thalassemia. PMID- 21617889 TI - Rosiglitazone improves spatial memory and decreases insoluble Abeta(1-42) in APP/PS1 mice. PMID- 21617891 TI - Treatment of motor fluctuations in Parkinson's disease. PMID- 21617890 TI - FMR1 premutation and full mutation molecular mechanisms related to autism. AB - Fragile X syndrome (FXS) is caused by an expanded CGG repeat (>200 repeats) in the 5' un-translated portion of the fragile X mental retardation 1 gene (FMR1) leading to a deficiency or absence of the FMR1 protein (FMRP). FMRP is an RNA binding protein that regulates the translation of a number of other genes that are important for synaptic development and plasticity. Furthermore, many of these genes, when mutated, have been linked to autism in the general population, which may explain the high comorbidity that exists between FXS and autism spectrum disorders (ASD). Additionally, premutation repeat expansions (55 to 200 CGG repeats) may also give rise to ASD through a different molecular mechanism that involves a direct toxic effect of FMR1 mRNA. It is believed that RNA toxicity underlies much of the premutation-related involvement, including developmental concerns like autism, as well as neurodegenerative issues with aging such as the fragile X-associated tremor ataxia syndrome (FXTAS). RNA toxicity can also lead to mitochondrial dysfunction, which is common in older premutation carriers both with and without FXTAS. Many of the problems with cellular dysregulation in both premutation and full mutation neurons also parallel the cellular abnormalities that have been documented in idiopathic autism. Research regarding dysregulation of neurotransmitter systems caused by the lack of FMRP in FXS, including metabotropic glutamate receptor 1/5 (mGluR1/5) pathway and GABA pathways, has led to new targeted treatments for FXS. Preliminary evidence suggests that these new targeted treatments will also be beneficial in non-fragile X forms of autism. PMID- 21617892 TI - Kynurenines and intestinal neurotransmission: the role of N-methyl-D-aspartate receptors. AB - Gastrointestinal neuroprotection involves the net effect of many mechanisms which protect the enteral nervous system and its cells from death, dysfunction or degeneration. Neuroprotection is also a therapeutic strategy, aimed at slowing or halting the progression of primary neuronal loss following acute or chronic diseases. The neuroprotective properties of a compound clearly have implications for an understanding of the mechanism of dysfunctions and for therapeutic approaches in a number of gastrointestinal diseases.This paper focused on the roles of glutamate and N-methyl-D-aspartate (NMDA) receptors in the intrinsic neuronal control of gastrointestinal motility; the consequences of inflammation on gastrointestinal motility changes; and the involvement of tryptophan metabolites (especially kynurenic acid) in the regulatory function of the enteral nervous system and the modulation of the inflammatory response. Common features in the mechanisms of action, illustrative evidence from animal models, and experimental neuroprotective therapies making use of the currently available possibilities are also discussed.Overall, the evidence suggests that gastrointestinal neuroprotection against inflammation and glutamate-induced neurotoxicity may be mediated synergistically through the blockade of NMDA receptors and the inhibition of neuronal nitric oxide synthase activity and xanthine oxidoreductase-dependent superoxide production. These components are likewise significant factors in the pathomechanism of gastrointestinal inflammatory diseases and inflammation-linked motility alterations. Inhibition of the enteric NMDA receptors by kynurenic acid or its analogues may provide a novel option via which to influence intestinal hypermotility and inflammatory processes simultaneously. PMID- 21617893 TI - The added value of contrast-enhanced ultrasound in patients with colorectal cancer undergoing preoperative evaluation with extensive gadobenate dimeglumine liver MRI. AB - OBJECTIVES: To evaluate the added value of pre- and intraoperative contrast enhanced ultrasound (transabdominal, or TCEUS and intraoperative, or ICEUS, respectively) in patients with known or highly suspected colorectal cancer liver metastases (CRLM) who have previously undergone extensive gadobenate dimeglumine (Gd-BOPTA) liver MRI. METHODS: Fifteen patients with a total of 31 lesions were included in the comparison of TCEUS vs. MRI and nine patients with a total of 19 lesions were included in the comparison of ICEUS vs. MRI. MRI examinations were performed before TCEUS and ICEUS. The analysis was performed lesion by lesion. Sensitivity, positive predictive value (PPV) and accuracy were calculated and compared. RESULTS: On comparing TCEUS with MRI, sensitivity differed significantly, with values of 87% and 100%, respectively (p value < 0.05), but there was no significant difference in PPV and accuracy. The comparison of ICEUS with MRI, however, showed no significant difference in sensitivity, PPV or accuracy. CONCLUSIONS: Transabdominal and intraoperative contrast-enhanced ultrasound have no added value in the preoperative evaluation of patients with CRLM undergoing extensive gadobenate dimeglumine liver MRI. PMID- 21617894 TI - Do older athletes reach limits in their performance during marathon running? AB - In the last decades, the participation of elderly trained people in endurance events such as marathon running has dramatically increased. Previous studies suggested that the performance of master runners (>40 years) during marathon running has improved. The aims of the study were (1) to analyze the changes in participation and performance trends of master marathon runners between 1980 and 2009, and (2) to compare the gender differences in performance as a function of age across the years. Running times of the best male and female runners between 20 and 79 years of age who competed in the New York City Marathon were analyzed. Gender differences in performance times were analyzed for the top 10 male and female runners between 20 and 65 years of age. The participation of master runners increased during the 1980-2009 period, to a greater extent for females compared to males. During that period, running times of master runners significantly (P < 0.01) decreased for males older than 64 years and for females older than 44 years, respectively. Gender differences in running times decreased over the last three decades but remained relatively stable across the ages during the last decade. These data suggest that male (>=65 years) and female (>=45 years) master runners have probably not yet reached their limits in marathon performance. The relative stability of gender differences in marathon running times across the different age groups over the last decade also suggests that age related declines in physiological function do not differ between male and female marathoners. PMID- 21617895 TI - Succession in stream biofilms is an environmentally driven gradient of stress tolerance. AB - The century-long research on succession has bestowed us with a number of theories, but little agreement on what causes species replacements through time. The majority of studies has explored the temporal trends of individual species in plant and much less so in microbial communities, arguing that interspecific interactions, especially competition, play a key role in community organization throughout succession. In this experimental investigation of periphytic succession in re-circulating laboratory streams, we examined the density and the relative abundance of diatoms and soft algae for 35 days across gradients of low to high nutrient supply (nitrogen + phosphorus) and low to intermediate current velocity (10 vs. 30 cm.s(-1)). All algal species were classified into trophic groups and morphological guilds, both of which responded more strongly to nutrient than current velocity manipulations, as shown by regression analyses. We concluded that within the manipulated environmental ranges: (1) Succession was a gradient of stress tolerance, driven primarily by nutrient supply and secondarily, by current velocity. Nutrient supply had a qualitative effect in determining whether the contribution of species tolerant vs. sensitive to nutrient limitation would increase through time, while current velocity had a quantitative influence and affected only the rate of this increase. (2) The mechanism of algal succession at a functional level was a neutral coexistence, whereby the tolerant low profile guild maintained high density when overgrown by sensitive species, while sensitive species, constituting mostly the motile and high profile guilds, were neither facilitated nor inhibited by tolerant species but controlled by the environment. It is suggested that the mechanism of succession may depend on the level of biological organization with interspecific interactions giving way to neutral coexistence along the hierarchy from species to functional groups. Considering that the functional makeup is strictly environmentally defined, while species composition reflects local and regional species pools that may exhibit substantial geographic variability, succession is deterministic at a functional level but stochastic at a species level. PMID- 21617896 TI - Terrestrial sources homogenize bacterial water quality during rainfall in two urbanized watersheds in Santa Barbara, CA. AB - Microbiological contamination from runoff is a human health concern in urbanized coastal environments, but the contamination sources are often unknown. This study quantified fecal indicator bacteria and compared the distributions of human specific genetic markers and bacterial community composition during dry and wet weather in urban creeks draining two neighboring watersheds in Santa Barbara, CA. In a prior study conducted during exclusively dry weather, the creeks were contaminated with human waste as indicated by elevated numbers of the human specific Bacteroidales marker HF183 (Sercu et al. in Environ Sci Technol 43:293 298, 2009). During the storm, fecal indicator bacterial numbers and loads increased orders of magnitude above dry weather conditions. Moreover, bacterial community composition drastically changed during rainfall and differed from dry weather flow by (1) increased bacterial diversity, (2) reduced spatial heterogeneity within and between watersheds, and (3) clone library sequences more related to terrestrial than freshwater taxa. Finally, the spatial patterns of human-associated genetic markers (HF183 and Methanobrevibacter smithii nifH gene) changed during wet weather, and the contribution of surface soils to M. smithii nifH gene detection was suspected. The increased fecal indicator bacteria numbers during wet weather were likely associated with terrestrial sources, instead of human waste sources that dominated during dry weather flow. PMID- 21617897 TI - Construction of an integrated map of Haliotis diversicolor using microsatellite markers. AB - Small abalone, Haliotis diversicolor, is naturally distributed along the coastal waters of East Asia from Japan to the Philippines. It is an economically important maricultured species in southern China and Taiwan. Genetic linkage maps for small abalone were constructed using a total of 308 simple sequence repeat markers including 297 novel markers. Segregation data on 96 progeny were genotyped using a pseudo-testcross strategy. Sixteen linkage groups were identified in both female and male maps, consistent with the haploid chromosome number. The female linkage map covered 758.3 cM, with an average interval of 5.2 cM. The male linkage map spanned a total genetic distance of 676.2 cM, with an average interval of 4.5 cM. An integrated map was constructed by incorporating the homologous parental linkage groups, resulting in 16 linkage groups with a total of 762.1 cM. Genome coverage of the integrated linkage map was approximately 80.7%. The genetic linkage maps of small abalone may facilitate marker-assisted selection and quantitative trait loci mapping. PMID- 21617898 TI - Effects of sucrose and trehalose on stability, kinetic properties, and thermal aggregation of firefly luciferase. AB - In this study, we used sugars as stabilizing additives to improve the thermostability and to inhibit aggregation of firefly luciferase. The combination of sucrose and trehalose has a strong stabilizing effect on firefly luciferase activity and prevents its thermoinactivation. These additives can also increase optimum temperature. It has been shown that the presence of both sucrose and trehalose can inhibit thermal aggregation of firefly luciferase and decrease bioluminescence decay rate. In order to understand the molecular mechanism of thermostabilization, we investigated the effects of sucrose and trehalose combination on the secondary structure of luciferase by Fourier transform infrared spectroscopy. Minor changes in content of secondary structure of firefly luciferase are observed upon treatment with additives. PMID- 21617899 TI - Implicit attenuation of subsequent emotion by cognitive activity. AB - The successful regulation of emotion relies heavily on executive function. Until very recently, however, the specific effects of executive function activity on other processes havereceived relatively little empirical attention.The link between executive function and emotional responses suggests that the activation of executive function may play an important role in the attenuation of emotional responses. We conducted twoexperiments to test the hypothesis that decrements in emotional responses following cognitive task performance might be caused, at least in part, by the activation of executive function.First, we sought to test whetherthe activation of executive function reducedemotional responsesimplicitly, without any volitional effort.Next, we sought to examine the link between prior activation of executivefunction and the attenuation of subsequent emotional responses, by comparing the effects of an executive-function-demanding task with the effects of a task that is equally effortful but does not engage executive function.We discuss our results in the context of the role of executive function in the attenuation of emotion. PMID- 21617900 TI - Up-regulation of thromboxane A2 impairs cerebrovascular eNOS function in aging atherosclerotic mice. AB - We previously reported that in healthy mouse cerebral arteries, endothelial nitric oxide synthase (eNOS) produces H2O2, leading to endothelium-dependent dilation. In contrast, thromboxane A2 (TXA2), a potent pro-oxidant and pro inflammatory endogenous vasoconstrictor, is associated with eNOS dysfunction. Our objectives were to elucidate whether (1) the cerebrovascular eNOS-H2O2 pathway was sensitive to oxidative stress associated with aging and dyslipidemia and (2) TXA2 contributed to cerebral eNOS dysfunction. Atherosclerotic (ATX = LDLR(-/-); hApoB(+/+)) and wild-type (WT) control mice were used at 3 and 12 months old (m/o). Three-m/o ATX mice were treated with the cardio-protective polyphenol catechin for 9 months. Dilations to ACh and the simultaneous eNOS-derived H2O2 production were recorded in isolated pressurized cerebral arteries. The age associated decrease in cerebral eNOS-H2O2 pathway observed in WT was premature in ATX mice, decreasing at 3 m/o and abolished at 12 m/o. Thromboxane synthase inhibition by furegrelate increased dilations at 12 months in WT and at 3 and 12 months in ATX mice, suggesting an anti-dilatory role of TXA2 with age hastened by dyslipidemia. In addition, the non-selective NADP(H) oxidase inhibitor apocynin improved the eNOS-H2O2 pathway only in 12-m/o ATX mice. Catechin normalized the function of this pathway, which became sensitive to L-NNA and insensitive to furegrelate or apocynin; catechin also prevented the rise in TXA2 synthase expression. In conclusion, the age-dependent cerebral endothelial dysfunction is precocious in dyslipidemia and involves TXA2 production that limits eNOS activity. Preventive catechin treatment reduced the impact of endogenous TXA2 on the control of cerebral tone and maintained eNOS function. PMID- 21617901 TI - Unusual renal manifestations in a girl with hyperthyroidism. AB - A 16-year-girl, a known case of hyperthyroidism since last 6 months, presented to the emergency with complaints of acute gastroenteritis of 5 days duration. At admission the child was found to be hypovolemic with acidotic breathing, which was thought to be due to dehydration. However, despite fluid correction she remained acidotic. A diagnosis of distal renal tubular acidosis (RTA) was suspected when her renal scan, for the cause of metabolic acidosis, revealed nephrocalcinosis. This was later confirmed by relevant investigations. She was started on treatment for distal RTA on which she symptomatically improved. The association of distal RTA and thyroid disorders is rarely reported in children. Till date there is only one report of distal RTA and hypothyroidism, but none with hyperthyroidism in this age group. The authors chose to report this case to highlight the fact that one should have a high index of suspicion for renal tubular disorders in children with thyroid hormone abnormalities, as these are treatable conditions if diagnosed early. PMID- 21617902 TI - Spontaneous pneumopericardium in a healthy full-term neonate. AB - Pneumopericardium is rare in neonates and might be of serious clinical significance. Neonatal pneumopericardium is usually a complication of mechanical ventilation in premature infants with respiratory distress syndrome. Here, the authors report a full-term neonate who developed spontaneous pneumopericardium which resolved spontaneously. PMID- 21617903 TI - Intrascrotal extratesticular neurofibroma as a possible cause of failed descent in ipsilateral testis. AB - Intrascrotal extratesticular neurofibromas (IEN) often originate from genitofemoral nerve (GFN) and present as a paratesticular mass. Synchronous presence of IEN and undescended testis has not been reported previously. A 12 year-old boy with neurocutaneous syndrome and congenital giant melanocytic nevi along with IEN and ipsilateral undescended testis is presented, to discuss the underlying pathophysiology of failed testicular descent in the presence of IEN. PMID- 21617904 TI - Symptomatic hypoglycemia causing brain injury in a term breast fed newborn following early discharge. AB - Cerebral metabolism and functioning depends upon an adequate blood glucose supply which provides for majority of the brain's energy requirement. Studies from the past have shown that neonatal hypoglycemia is associated with acute and long term neurological sequelae. Early discharge without adequately established breast feeding may lead to feeding problems, post discharge hypoglycemia and its associated neurological complications. The authors describe one such case of an exclusively breast fed term newborn who presented on day 3 with symptomatic hypoglycemia and associated neurological injury. PMID- 21617906 TI - Hematidrosis and hemolacria: a case report. AB - Hematidrosis (bloody sweat) and hemolacria (bloody tears) are very rare phenomena. The authors herein report hematidrosis and hemolacria occurring together in a 10-year-old girl. The role of propranolol in its treatment is highlighted. PMID- 21617905 TI - Factors affecting duration of chronic fatigue syndrome in pediatric patients. AB - OBJECTIVE: To determine factors affecting duration of chronic fatigue syndrome (CFS) in pediatric patients. METHODS: This Retrospective cohort consisted of patients with CFS at the regional referral infectious disease clinic for evaluation of fatigue in children and adolescents. Demographic, clinical, and laboratory data were analyzed to identify the impact on duration and severity of pediatric CFS. RESULTS: A total number of 53 predominantly white (98.1%) patients with CFS, aged 9-18 years, were included in the study. Other than fatigue, headaches and sleep disturbance were the most common symptoms of pediatric CFS. Seropositive status for Borrelia burgdorferi (B. burgdorferi) and Epstein-Barr virus (EBV) was identified in 66% of the patients with the diagnosis of CFS by CDC criteria. No association was found between the CFS symptoms, gender, or age at diagnosis and duration of fatigue symptoms. Duration of CFS was associated with high Body-Mass Index (BMI) in a regression model after adjustment for patient's age, gender, and seropositive status for B. burgdorferi and/or EBV (0.34 +/- 0.15, P < 0.04). CONCLUSIONS: BMI is significantly associated with prolonged duration of CFS. PMID- 21617907 TI - Validation of Pediatric Index of Mortality 2 in three pediatric intensive care units in Hong Kong. AB - OBJECTIVE: To compare the probabilities of death predicted by Pediatric Index of Mortality 2 (PIM2) and Pediatric Index of Mortality 1 (PIM1) models against actual mortalities in 3 pediatric intensive care units in Hong Kong. METHODS: All consecutive patients admitted to pediatric intensive care units in 3 regional non teaching hospitals between Jan 2006 and December 2006 were included. The data required for PIM1 and PIM2 were collected. The probabilities of death predicted by PIM1 and PIM2 were validated against the actual probability of mortality. RESULTS: Of the 540 patients were included in this study, only seven deaths were observed (1.3%). PIM 1 and PIM 2 overestimated the mortality rate by giving a greater mortality rate. (PIM1: 13.4, Standardized mortality ratio = 0.52, 95% CI = 0.14 to 0.91; PIM2: 14.2, Standardized mortality ratio =0.49, 95% CI = 0.13 to 0.86) The discrimination of PIM1 and PIM2 were satisfactory as reflected by area under receiver-operator characteristic curve of 0.889 (95% CI: 0.703 to 1.000) and 0.904 (95% CI: 0.738 to 1.000) respectively. Calibration was not possible due to insufficient death cases. CONCLUSIONS: The current study showed that PIM2 and PIM1 had similar accuracy in mortality prediction in Hong Kong. The current study also demonstrated the difficulty to calibrate the mortality model when actual mortality rate is low. PMID- 21617908 TI - A primary care approach for adolescent care and counseling services. AB - OBJECTIVE: Adolescents can have mental, emotional, and behavior problems that are a source of stress for the child as well as the family, school and community. These may disrupt the adolescent's ability to function normally. Adolescents also have reproductive concerns especially at menarche. Considering the extent of problems of adolescents and the lack of adolescent care and counseling services, it was felt that community adolescent care counseling services should be made available. This article describes the steps involved in the setting up of Taluk model of adolescent care and counseling services. METHODS: Following steps were involved in setting up a Taluk model of adolescent care counseling service delivery system. Step I: Focus Group Discussions (FGDs) among Stakeholders. Step II: Conceptualization and Strategy planning for service delivery. III: Finalization of service delivery model Step IV: Workshops for finalization of TSQ T 2008 version the tool to be used for assessing the adolescents in the ARSH clinics. Step V: Training Programme for Medical/Paramedical health staff. Step VI: Awareness programs for mothers of adolescents. Step VII: Setting up of ACS/ARSH clinics at Taluk hospitals. Step VIII: Evaluation of the utilization of services at Taluk hospitals. The clinic has been well utilized with 1,588 adolescents being seen in 2 years. RESULTS: Medical and Reproductive problems among adolescent girls were anemia, underweight, dysmenorrhoea, menstrual irregularities and symptoms of Polycystic Ovarian Syndrome, whereas among boys problems were mostly related to concerns about masturbation and its perceived ill effects. The psychosocial problems ranged from minor anxieties, sadness and adjustment problems to psychiatric disorders. Scholastic problems included poor concentration, poor study habits and low intelligence quotient. CONCLUSIONS: The success of the clinics in these five hospitals can be replicated in other parts of the state as well as the country. These will go a long way to ameliorate the morbidity of adolescents. PMID- 21617909 TI - Effectiveness of a reproductive sexual health education package among school going adolescents. AB - OBJECTIVE: To assess the effectiveness of a school based "Adolescent Reproductive Sexual Health Education (ARSHE) Package" in improving students' knowledge on reproductive sexual health matters. METHODS: An ARSHE package originally developed at Child Development Centre, Kerala, modified and approved by ICMR taskforce group was administered in three urban schools (One boys only, one girls only and one co-education) and one co-education rural school at Thiruvananthapuram district, Kerala. The study sample consisted of 1,586 adolescents including 996 boys and 560 girls of class IX and XI. Pre and post intervention knowledge regarding reproductive sexual health matters was assessed using a self-administered questionnaire. RESULTS: In the pre-intervention period, it was observed that majority of adolescents were poorly informed about reproductive sexual health matters, particularly about contraceptives. As compared to boys, girls had much poorer knowledge about prevention of pregnancy and after intervention; there was a statistically significant increase in the knowledge in both boys and girls. Among girls percentage of poor knowledge had reduced significantly from 64.1% to 8.3% and among boys from 37.7% to 3.5%. Similarly, increase in knowledge level was also observed in various other aspects of reproductive and sexual health including, STI, HIV/AIDS and perceptions about premarital sex. CONCLUSIONS: The study results revealed the feasibility and effectiveness of school based reproductive and sexual health education intervention programs for adolescents. PMID- 21617910 TI - Priority mental health disorders of children and adolescents in primary-care pediatric settings in India 4: training and capacity building. AB - Training in the primary-care child and adolescent mental health should take into consideration the local milieu, national health care education and development. It should aim to improve the mental health knowledge, competency as well as develop professional relationships between various primary, secondary and tertiary-care mental health providers to enhance outcomes. The collaborative training between the various stakeholders in the Child and Adolescent Mental Health (CAMH) should be enhanced. Currently, the favoured methods, to augment the training for practicing Primary-care Physicians, like CME and short training programs with their specific goals, settings and methodology are well documented. However, to improve the skills in CAMH for medical trainees at undergraduate and postgraduate levels, restructuring of the curriculum is essential. PMID- 21617911 TI - Priority mental health disorders of children and adolescents in primary-care pediatric settings in India 3: psychotherapy and other non-pharmacological interventions. AB - The pediatrician is a primary column of support for children and adolescents with a myriad of mental health problems in low-mental health care resource countries like India. While majority of mental health consultations happen in primary-care, and only 10% are referred successfully for specialised help, there is a clear role for pediatrician psychotherapists in primary care. The primary-care pediatricians should be aware of the indications for psychotherapy, the various approaches that could be used in primary-care settings, the structure and the process of the psychotherapeutic technique involved, the suggested specific techniques for the Priority Mental Health Disorders and the evidence available to support their use as well as the developmental modifications that are required based on the cognitive development of the child or adolescent. PMID- 21617912 TI - Priority mental health disorders of children and adolescents in primary-care pediatric settings in India 2: diagnosis, pharmacological treatment and referral. AB - The advent of pediatric psychopharmacology has enormously improved psychiatric care of children and adolescents. Nonetheless, our practice of diagnosis, treatment and referral in primary-care pediatric settings is not optimum as current evidence based knowledge is not regularly applied in the actual clinical circumstances. To help primary-care pediatricians minimise this in research clinical practice, pharmacological treatment and referral in their clinical practice, they need to follow a two-tier diagnostic and multi axial treatment approach. The two-tier diagnostic approach of using a screening measure followed by confirmation of the screen positive cases with reference standard clinical criterion, improves the sensitivity and specificity. The multiaxial treatment has the advantage of offering a holistic approach to the intervention and improve prognosis from the interacting axes. The primary-care physician should be aware of the medications of choice for the Priority Mental Health Disorders and their drug interactions. Finally, referral of cases with atypical presentations, multiple comorbidities and poor response to the first-line of treatment needs referral to the next tier in the system. PMID- 21617913 TI - HIF-1 is involved in high glucose-induced paracellular permeability of brain endothelial cells. AB - Experimental evidence from human patients and animal models of diabetes has demonstrated that hyperglycemia increases blood-brain barrier (BBB) permeability, which is associated with increased risk of neurological dysfunction. However, the mechanism underlying high glucose-induced BBB disruption is not understood. Here we investigated the role of hypoxia-inducible factor-1 (HIF-1) in high glucose induced endothelial permeability in vitro using mouse brain microvascular endothelial cells (b.End3). Our results demonstrated that high glucose (30 mM) upregulated the protein level of HIF-1alpha, the regulatable subunit of HIF-1, and increased the transcriptional activity of HIF-1 in the endothelial cells. At the same time, high glucose increased the paracellular permeability associated with diminished expression and disrupted continuity of tight junction proteins occludin and zona occludens protein-1 (ZO-1) of the endothelial cells. Upregulating HIF-1 activity by cobalt chloride increased the paracellular permeability of the endothelial cells exposed to normal glucose (5.5 mM). In contrast, downregulating HIF-1 activity by HIF-1alpha inhibitors and HIF-1alpha specific siRNA ameliorated the increased paracellular permeability and the alterations of distribution pattern of occludin and ZO-1 induced by high glucose. In addition, high glucose increased expression of vascular endothelial growth factor (VEGF), a downstream gene of HIF-1. Inhibiting VEGF improved the expression pattern of occludin and ZO-1, and attenuated the endothelial leakage. Furthermore, key results were confirmed in human brain microvascular endothelial cells. These results strongly indicate that HIF-1 plays an important role in high glucose-induced BBB dysfunction. The results will help us understand the molecular mechanisms involved in hyperglycemia-induced BBB dysfunction and neurological outcomes. PMID- 21617914 TI - A placebo-controlled study of sertraline's effect on cortisol response to the dexamethasone/corticotropin-releasing hormone test in healthy adults. AB - RATIONALE: The dexamethasone/corticotropin-releasing hormone (Dex/CRH) test is a neuroendocrine probe involving serial blood sampling of cortisol during a standardized pharmacological challenge without inducing psychological distress in humans. Some past studies in depressed patients have shown a "normalization" or decrease in cortisol response to the Dex/CRH test following successful treatment with an antidepressant. Studies in nondepressed healthy adult samples have also shown aberrant cortisol reactivity to be associated with depression risk factors. These findings prompted research into the use of the Dex/CRH test as a tool for developing antidepressant drugs. OBJECTIVES: In this study, the Dex/CRH test was evaluated with regard to its potential utility for drug development in nonclinical samples. METHODS: The Dex/CRH test was administered before and after 6 weeks of blinded treatment with either sertraline 100 mg/day or matching placebo in 22 healthy adults (13 women, nine men). RESULTS: Cortisol response to the Dex/CRH test increased following treatment with standard doses of sertraline, compared to placebo, after controlling for age and sex. CONCLUSIONS: The observed pattern of change contrasts with results from published studies in depressed patients and with our initial hypothesis. PMID- 21617915 TI - Genetic testing in renal disease. AB - A revolution is happening in genetics! The decoding of the first genome in 2003 was a large international collaborative effort that took about 13 years at a cost of around $2.7 billion. Now, only a few years later, new technology allows the sequencing of an entire genome within a few weeks--and at a cost of less than $10,000. The vaunted $1000 genome is within reach. These extraordinary advances will undoubtedly transform the way we practice medicine. But, like any new technology, it carries risks, as well as benefits. As physicians, we need to understand the implications in order to best utilise these advances for our patients and to provide informed advice. In this review, our aim is to explain these new technologies, to separate the hype from the reality and to address some of the resulting questions and implications. The practical objective is to provide a simple overview of the available technologies and of purpose to which they are best suited. PMID- 21617916 TI - Maternal smoking during pregnancy and kidney volume in the offspring: the Generation R Study. AB - An adverse fetal environment leads to smaller kidneys, with fewer nephrons, which might predispose an individual to the development of kidney disease and hypertension in adult life. In a prospective cohort study among 1,072 children followed from early fetal life onward, we examined whether maternal smoking during pregnancy, as a significant adverse fetal exposure, is associated with fetal (third trimester of pregnancy, n = 1,031) and infant kidney volume (2 years of age, n = 538) measured by ultrasound. Analyses were adjusted for various potential confounders. Among mothers who continued smoking, we observed dose dependent associations between the number of cigarettes smoked during pregnancy and kidney volume in fetal life. Smoking less than five cigarettes per day was associated with larger fetal combined kidney volume, while smoking more than ten cigarettes per day tended to be associated with smaller fetal combined kidney volume (p for trend: 0.002). This pattern was not significant for kidney volume at the age of 2 years. Our results suggest that smoking during pregnancy might affect kidney development in fetal life with a dose-dependent relationship. Further studies are needed to assess the underlying mechanisms and whether these differences in fetal kidney volume have postnatal consequences for kidney function and blood pressure. PMID- 21617917 TI - PIK3CA mutations rarely demonstrate genotypic intratumoral heterogeneity and are selected for in breast cancer progression. AB - PIK3CA gene mutations are the most common activating mutations in human breast cancer. Its association with hormone receptor-positive breast cancer makes it a prime target for clinical therapeutic advances to maintain anti-estrogen responsiveness. In anticipation of this therapeutic approach, we have evaluated intratumoral heterogeneity in primary breast cancers with regard to PIK3CA mutation status. In addition, we have assessed for the presence of the mutation in paired pre-invasive breast cancer and metastases. To assess for intratumoral heterogeneity, separate tumor blocks from primary breast cancers (n = 63) were genotyped for PIK3CA mutations. Available paired tissue samples from breast tumors known to harbor mutations underwent massARRAY genotyping (n = 70) to identify PIK3CA and AKT1(E17K) mutations. Cores were macro-dissected from matched tissue, including normal breast, benign lymph nodes (LN), ductal carcinoma in situ, regional LN metastases, and distant metastases. Matched samples underwent genetic fingerprinting by multiple SNP genotyping to confirm genetic identity. Intratumoral heterogeneity is minimal with a concordance rate of 95.2% between two different blocks from primary breast cancers. Complete concordance of PIK3CA mutations is noted between primary breast cancer and DCIS. PIK3CA mutations in primary breast cancer are detected in matched regional LNs (91.7%) and distant metastases (100%). Mutation detection by massARRAY genotyping is sensitive but may be affected by sample quality. Intratumoral heterogeneity as measured by PIK3CA genotype is rare; PIK3CA mutations occur early and are selected for in breast cancer progression. HapMap analysis is an essential control for paired sample analysis. This data is clinically important, particularly, for the design of therapies targeting the PI3K/AKT pathway, as it offers confidence that the detection of PIK3CA mutations in the invasive primary tumor will accurately reflect breast cancer biology. PMID- 21617918 TI - Age-specific trends of survival in metastatic breast cancer: 26 years longitudinal data from a population-based cancer registry in Stockholm, Sweden. AB - Treatment of metastatic breast cancer (MBC) has evolved during the last decades but it is largely unknown whether this has led to improved survival in the general MBC population. Based on the regional, population-based breast cancer registry, we identified 5,463 patients diagnosed with MBC in Stockholm County during 1979-2004. Patients were divided into five cohorts based on the year of first MBC diagnosis and observed and relative survival were compared across the cohorts after adjustment for potential confounders. A significant trend of better survival over time was demonstrated for patients 60 years or younger (P < 0.001, by log-rank test for trend), but not for older patients (P = 0.12) or for the whole MBC population (P = 0.13). The adjusted observed survival of patients <= 60 years was significantly improved in the 2000-2004 cohort (P < 0.001, hazard ratio = 0.7, 95% confidence interval = 0.58-0.84), corresponding to a clinically significant increase of median survival with more than 3 months and absolute increase of 5-year survival with 8% or more compared to previous periods. Similarly, relative survival analysis indicated a 31% decreased mortality for the younger subpopulation in the 2000-2004 cohort (P < 0.001). Systemic adjuvant treatment was a negative prognostic factor after distant recurrence. Treatment advancements in MBC are not reflected by better survival for the whole MBC population. An improvement is only observed after the year 2000 and is restricted to younger patients. PMID- 21617919 TI - Unmet supportive care needs: a cross-cultural comparison between Hong Kong Chinese and German Caucasian women with breast cancer. AB - The comparison of psychosocial needs across different cultural settings can identify cultural and service impacts on psychosocial outcomes. We compare psychosocial needs in Hong Kong Chinese and German Caucasian women with breast cancer. Completed questionnaires were collected from 348 Chinese and 292 German women with breast cancer for assessing unmet psychosocial needs (Supportive Care Needs Survey Short Form), psychological distress (the Hospital Anxiety and Depression scale), and listed physical and psychological symptoms. Only 11% of the participants reported not needing help for any of the 34 items. More German (14%) than Chinese women (8%) reported no unmet needs (chi(2) = 6.16, P = .013). With both samples combined, the Health System and Information domain unmet needs were the most prevalent, apart from one Psychological need domain item, "Fear about the cancer spreading." Chinese and German samples differed significantly in prevalence and patterns of unmet psychosocial needs. Multivariate adjustment for demographic, clinical, and sample characteristics, psychological distress, and symptoms showed that significantly greater unmet Health system and Information, and Patient care and support domain needs, associated with the presence of symptoms (beta = .232, P < .001), high HADS Anxiety (beta = .187, P < .001), higher education attainment (beta = .120, P = .002), and Chinese sample membership (beta = .280, P < .001). Greater unmet Psychological, Physical and Daily Living, and Sexuality domain needs were associated with the presence of symptoms, psychological distress, and German group membership, among others. German women reported more anxiety (t = 10.45, P < .001) and depression (t = 3.71, P < .001). In post hoc analyses, German, but not Chinese women reporting greater anxiety and depression had greater unmet Psychological and Sexuality domain needs (P < .001). It can be concluded that culture-specific differences in supportive care needs exist. Hong Kong Chinese women prioritize needs for information about their disease and treatment, whereas German Caucasian women prioritize physical and psychological support. Planning for cancer supportive care services or interventions to reduce unmet needs must consider cultural and/or health service contexts. PMID- 21617920 TI - Effect of false-positives and women's characteristics on long-term adherence to breast cancer screening. AB - False-positive results may influence adherence to mammography screening. The effectiveness of breast cancer screening is closely related to adequate adherence among the target population. The objective of this study was to evaluate how false-positives and women's characteristics affect the likelihood of reattendance at routine breast cancer screening in a sequence of routine screening invitations. We performed a retrospective cohort study of 1,371,218 women aged 45 69 years, eligible for the next routine screening, who underwent 4,545,346 screening mammograms from 1990 to 2006. We estimated the likelihood of attendance at seven sequential screening mammograms. Multilevel discrete time hazard models were used to estimate the effect of false-positive results on reattendance, and the odds ratios (OR) of non-attendance for the women's personal characteristics studied. The overall reattendance rate at the second screening was 81.7% while at the seventh screening was 95.6%. At the second screening invitation reattendance among women with and without a false-positive mammogram was 79.3 vs. 85.3%, respectively. At the fourth and seventh screenings, these percentages were 86.3 vs. 89.9% and 94.6 vs. 96.0%, respectively. The study variables associated with a higher risk of failing to participate in subsequent screenings were oldest age (OR = 8.48; 95% CI: 8.31-8.65), not attending their first screening invitation (OR = 1.12; 95% CI: 1.11-1.14), and previous invasive procedures (OR = 1.09; 95% CI: 1.07-1.10). The risk of non-attendance was lower in women with a familial history of breast cancer (OR = 0.97; 95% CI: 0.96-0.99), and those using hormone replacement therapy (OR = 0.96; 95% CI: 0.94-0.97). In conclusion, reattendance was lower in women with false-positive mammograms than in those with negative results, although this difference decreased with the number of completed screening participations, suggesting that abnormal results in earlier screenings more strongly influence behavior. These findings may be useful in providing women with accurate information and in improving the effectiveness of screening programs. PMID- 21617921 TI - Antioxidant prophylaxis for cellular injury in ovarian surface epithelium resulting from CO2 pneumoperitoneum in a laparoscopic rat model. AB - OBJECTIVE: Selective cytoprotective functions of vitamin E, N-acetyl-L: cysteine, and amifostine have been used as a preventer of ischemia injury by expelling the free oxygen radicals leading to stabilization of the cellular membranes. The purpose of this experimental study was to investigate the oxidative stress related to cellular injury in ovarian surface epithelium and the effect of prophylaxis with an anti-oxidant using laparoscopic rat model. DESIGN: Laparoscopic rat model. MATERIALS AND METHODS: Randomly allocated 40 Wistar Albino female rats have been used for the pneumoperitoneum model which was constituted to fix the intraabdominal pressure on 5 mmHg for 60 min. The antioxidants, vitamin E and NAC were given to rats 3 days before the operation and were applied for 30 days; amifostine was applied 30 min before the operation until after for 7 days. After abdominal desufflation, over biopsies were made on the 13th min, 24th h, and 7th and 30th days. By using of transmission electron microscopy, the damage on cells and organels were assessed and graded. RESULTS: In ovarian surface epithelium, the apical surface specializations were affected in all groups except Vit E group:The microvilli were irregular and coarse and had disappeared in some places. Some cells were separated from the epithelium. In addition, mitochondria degeneration was observed in all group except Vit E. CONCLUSIONS: In the early period of laparoscopy, reversible cellular damage occurs and this damage can be prevented by vitamin E. PMID- 21617922 TI - Minimal invasive treatment options in pregnant women with ovarian vein syndrome. AB - PURPOSE: To describe the treatment modalities of symptomatic ovarian vein syndrome in pregnancy. METHODS: In our study, we included 12 pregnant women with right ureter and kidney dilatation; caused by ureteric obstruction. In 11 out of 12, we insert a DJ stent and in one woman we applied percutaneous nephrostomy, because of intermittent abdominal pain, resistant on analgetic therapy or feverish pyelonephritis. DJ stents and nephrostomy were inserted under ultrasound guidance without anaesthesia. RESULTS: After insertion of DJ stents, respective percutaneous nephrostomy colic attacks went back immediately and the feverish pyelonephritis in few days. At no time during these procedures, there was a risk for pregnancy. CONCLUSION: Ovarian vein syndrome in pregnancy can lead to violent colic pain and can become complicated by accompanied pyelonephritis. In these cases insertion of a DJ stent or percutaneous nephrostomy under ultrasound guidance is possible and safe, and leads to an improvement of complaints immediately. PMID- 21617923 TI - Prolonged progression-free survival in patients with chronic lymphocytic leukemia receiving granulocyte colony-stimulating factor during treatment with fludarabine, cyclophosphamide, and rituximab. AB - The clinical benefit of the addition of granulocyte colony-stimulating factor (G CSF) to standard immunochemotherapy of chronic lymphocytic leukemia (CLL) with fludarabine, cyclophosphamide, and rituximab (FCR) is still unclear. In this retrospective study we analyzed the outcome of 32 consecutive patients with CLL during treatment with FCR. Sixteen patients received G-CSF for treatment of CTC grade 3 or 4 neutropenia or febrile neutropenia at some point during therapy and 16 did not. Both groups were well balanced for clinical and biological risk factors. Overall response rates were not significantly different (94% vs. 75%; p=0.144). Interestingly, a significantly better progression-free survival (100% vs. 35.4% at 24 months; p<0.001) and even overall survival (100% vs. 77.8% at 24 months; p=0.022) was observed in patients receiving G-CSF. While the underlying cause remains to be elucidated, these data strongly suggest an association of the addition of G-CSF to FCR therapy with final patient outcome. PMID- 21617924 TI - Reconstruction of long-term tobacco consumption trends in Australia and their relationship to lung cancer mortality. AB - OBJECTIVES: Analysis of long-term trends in smoking and causes of death in Australia are prevented by a lack of detailed tobacco consumption data prior to World War II. The objective of this study was to reconstruct data on tobacco consumption in Australia back to 1887 and examine its relationship with population-level lung cancer mortality, corrected for biases and miscoding. METHODS: Back-extrapolation techniques and existing tobacco sales data were combined to estimate tobacco consumption prior to the 1940s. The relationship of tobacco and lung cancer mortality was examined with descriptive period and cohort analyses and log-linear Poisson regression models of cumulative cohort consumption. RESULTS: The results show that tobacco consumption rose steadily in Australia for the majority of years from 1887 to World War II, before increasing drastically in the following years and then falling sharply to the present day. Lung cancer mortality was strongly influenced by tobacco consumption, peaking 20 25 years after the peak in tobacco consumption for men and 25-30 years for women. Regression models found cumulative consumption a very strong predictor of mortality. CONCLUSIONS: Period and cohort trends in smoking and lung cancer were similar to many other Western countries. The effectiveness of smoking reduction campaigns in Australia clearly reduced male lung cancer mortality and provides guidance for other countries, such as China, where smoking prevalence remains high. PMID- 21617925 TI - Putting a value on the avoidance of false positive results when screening for inherited metabolic disease in the newborn. AB - Despite the increase in the number of inherited metabolic diseases that can be detected at birth using a single dried blood spot sample, the impact of false positive results on parents remains a concern. We used an economic approach - the contingent valuation method - which asks parents to give their maximum willingness to pay for an extension in a screening programme and the degree to which the potential for false positive results diminishes their valuations. 160 parents of a child or children under the age of 16 years were surveyed and given descriptions of the current screening programme in the UK, an extended programme and an extended programme with no false positives. 148 (92.5%) respondents said they would accept the screen for the five extra conditions in an expanded screening programme whilst 10 (6.3%) said they would not and two were unsure. When asked to indicate if they would choose to be screened under an expanded screening programme with no false positive results, 152 (95%) said they would, five (3.1%) said they would not, two were unsure, and there was one non-response. 151 (94.4%) said they preferred the hypothetical test with no false-positives. The mean willingness to pay for the expanded programme was L178 compared to L219 for the hypothetical expanded programme without false positives (p > 0.05). The results suggest that there is widespread parental support for extended screening in the UK and that the number of false-positives is a relatively small issue. PMID- 21617926 TI - Synovium, synergy and serendipity. PMID- 21617927 TI - Prenatal screening of sialic acid storage disease and confirmation in cultured fibroblasts by LC-MS/MS. AB - Sialic acid storage disease (SASD) is an inborn error resulting from defects in the lysosomal membrane protein sialin. The SASD phenotypical spectrum ranges from a severe presentation, infantile sialic acid storage disease (ISSD) which may present as hydrops fetalis, to a relatively mild form, Salla disease. Screening for SASD is performed by determination of free sialic acid (FSA) in urine or amniotic fluid supernatant (AFS). Subsequent diagnosis of SASD is performed by quantification of FSA in cultured fibroblasts and by mutation analysis of the sialin gene, SLC17A5. We describe simple quantitative procedures to determine FSA as well as conjugated sialic acid in AFS, and FSA in cultured fibroblasts, using isotope dilution ((13)C(3)-sialic acid) and multiple reaction monitoring LC-ESI MS/MS. The whole procedure can be performed in 2-4 h. Reference values in AFS were 0-8.2 MUmol/L for 15-25 weeks of gestation and 3.2-12.0 MUmol/L for 26-38 weeks of gestation. In AFS samples from five fetuses affected with ISSD FSA was 23.9-58.9 MUmol/L demonstrating that this method is able to discriminate ISSD pregnancies from normal ones. The method was also validated for determination of FSA in fibroblast homogenates. FSA in SASD fibroblasts (ISSD; 20-154 nmol/mg protein, intermediate SASD; 12.9-15.1 nmol/mg, Salla disease; 5.9-7.4 nmol/mg) was clearly elevated compared to normal controls (0.3-2.2 nmol/mg). In conclusion, we report simple quantitative procedures to determine FSA in AFS and cultured fibroblasts improving both prenatal diagnostic efficacy for ISSD as well as confirmatory testing in cultured fibroblasts following initial screening in urine or AFS. PMID- 21617928 TI - Simultaneous production and characterization of medium-chain-length polyhydroxyalkanoates and alginate oligosaccharides by Pseudomonas mendocina NK 01. AB - When Pseudomonas mendocina NK-01 was cultivated in a 200-L fermentor using glucose as carbon source, 0.316 g L(-1) medium-chain-length polyhydroxyalkanoate (PHA(MCL)) and 0.57 g L(-1) alginate oligosaccharides (AO) were obtained at the end of the process. GC/MS was used to characterize the PHA(MCL), which was found to be a polymer mainly consisting of 3HO (3-hydroxyoctanoate) and 3HD (3 hydroxydecanoate). T (m) and T (g) values for the PHA(MCL) were 51.03 degrees C and -41.21 degrees C, respectively, by DSC. Its decomposition temperature was about 300 degrees C. The elongation at break was 700% under 12 MPa stress. MS and GPC were also carried out to characterize the AO which had weight-average molecular weights of 1,546 and 1,029 Da, respectively, for the two main components at the end of the fermentation process. MS analysis revealed that the AO were consisted of beta-D-mannuronic acid and/or alpha-L-guluronic acid, and the beta-D-mannuronic acid and/or alpha-L-guluronic acid residues were partially acetylated at position C2 or C3. PMID- 21617929 TI - Separation of ionic liquid [Mmim][DMP] and glucose from enzymatic hydrolysis mixture of cellulose using alumina column chromatography. AB - Pretreatment of cellulose with ionic liquids (ILs) can improve the efficiency of the hydrolysis by increasing the surface area of the substrates accessible to solvents and cellulases. However, the IL methods are facing challenges to separate the hydrolyzed sugar products as well as the renewable ILs from the complex hydrolysis mixtures. In this study, an alumina column chromatography (ACC) method was developed for the separation of hydrophilic IL N-methyl-N methylimidazolium dimethyl phosphate ([Mmim][DMP]) and glucose, which was the main ingredient of the monosaccharide hydrolyzate. The processing parameters involved in ACC separation were investigated in detail. Our results showed that the recovery yields of [Mmim][DMP] and glucose can reach up to 93.38% and 90.14%, respectively, under the optimized parameters: the sampling ratio of 1:20 between the applied sample volume and the bed volume of the column; a gradient elution using methanol (100%, 150 ml) and then water (170 ml) as eluents with 1 ml/min flow rate. The recovered [Mmim][DMP] showed qualified property and was effective in a new hydrolysis reaction. In addition, scale-up ACC separations were successfully done with satisfied separation performance. The results indicated that the ACC is one of the available methods for the separation of ILs and monosaccharides from the hydrolysis mixtures. PMID- 21617931 TI - A potential use of embryonic stem cell medium for the in vitro culture of preimplantation embryos. AB - PURPOSE: To assess the impact of embryonic stem cell culture medium (ESCM) on the pre- and post-implantation development of the mouse embryo, as a mammalian model, in comparison with the conventional culture medium, a potassium simplex optimized medium (KSOM). METHODS: Development in ESCM versus KSOM was compared in terms of embryo morphology, cleavage, cavitation, hatching, cell number, expression of TE and ICM transcription factors (Cdx2 and Oct4, respectively), implantation, and development in utero. RESULTS: An enriched medium like ESCM can be beneficial for in vitro embryo development when cultured from the 8-cell stage, as evidenced by promotion of blastocyst development with respect to cavity expansion, hatching, and cell division. Such benefits were not observed when embryos were cultured from the 2-cell stage. CONCLUSIONS: ESCM may augment in vitro embryo development from the 8-cell stage. Using different culture media at different stages may be beneficial to achieve more effective human in vitro fertilization. PMID- 21617930 TI - The contribution of mitochondrial function to reproductive aging. AB - PURPOSE: The number of women attempting to conceive between the ages of 36 and 44 has increased significantly in the last decade. While it is well established that women's reproductive success dramatically declines with age, the underlying physiological changes responsible for this phenomenon are not well understood. With assisted reproductive technologies, it is clear that oocyte quality is a likely cause since women over 40 undergoing in vitro fertilization (IVF) with oocytes donated by younger women have success rates comparable to young patients. Apart from oocyte donation, there is no known intervention to improve the pregnancy outcome of older patients. The aim of this paper was the review the relevant data on the potential role of mitochondria in reproductive aging. METHOD: Review of current literature on the subject. RESULTS: We present the current evidence that associate mitochondrial dysfunction with age related decrease in female reproductive outcome. CONCLUSIONS: The aging process is complex, driven by a multitude of factors thought to modulate cellular and organism life span. Although the factors responsible for diminished oocyte quality remain to be elucidated, the present review focuses on the potential role of impaired mitochondrial function. PMID- 21617932 TI - Umbilical cord blood banking: an update. AB - BACKGROUND: Umbilical cord blood is a potential vast source of primitive hematopoietic stem and progenitor cells available for clinical application to reconstitute the hematopoietic system and/or restore immunological function in affected individuals requiring treatment. Cord blood can be used as an alternative source for bone marrow transplantation and its use is developing into a new field of treatment for pediatric and adult patients presenting with hematological disorders, immunological defects and specific genetic diseases. DISCUSSION: More than 25,000 allogeneic cord blood transplantations have been performed worldwide since the first cord blood transplantation in 1988. There are two banking options for storing umbilical cord blood [private (family) and public]. Cord blood stored in private banks are used for either autologous or allogeneic transplants for the infant donor or related family members but private cord blood banks are not searchable or available to the public. More than 780,000 cord blood units are stored in over 130 private cord blood banks, worldwide, and over 400,000 units in more than 100 quality controlled public cord blood banks. CONCLUSIONS: Researchers continue to evaluate the usefulness of cord blood cells in treating human diseases or disorders for purposes other than hematological disorders including heart disease, strokes, brain or spinal cord injuries and cancer. This review summarizes the status of umbilical cord blood banking, its history and current and potential use in the treatment of human disease. PMID- 21617933 TI - Does medial rotational deformity of the whole pelvis universally exist in unilateral DDH? AB - INTRODUCTION: There are no reports on the morphologic features of the entire pelvis in the subgroup of dislocation of hips according to the classification system of Tonnis. In addition, the correlation of excessive medial rotation of the lower pelvis with the increased acetabular anteversion was uncertain in the children with unilateral developmental dysplasia of the hip (DDH). PATIENTS AND METHODS: In the study, 74 patients with unilateral DDH and 31 subjects of normal control were involved, and the images of three-dimensional computed tomography (3D-CT) were retrospectively reconstructed to compare the acetabular anteversion angle (AA), the rotational angle of the upper pelvis (URA), and the rotational angle of the lower pelvis (LRA). The correlation of the AA with LRA was analyzed. RESULTS: In the group of grade II and IV, though the LRA was increased on the dislocated side as compared with the unaffected side (P < 0.05), the URA did not differ between the two sides (P > 0.05). There was positive correlation between AA and LRA on the dislocated side in the group of grade II, III, and IV. CONCLUSION: The excessive medial rotation of whole pelvis on the dislocated side was not universally presented in unilateral DDH. The excessive medial rotational deformity of the lower pelvis could induce increased acetabular anteversion. An individualized treatment plan based on the accurate assessment of morphologic features of the whole pelvis and the exact understanding for underlying causes of acetabular anteversion through 3D-CT should be considered to avoid complications during surgery. PMID- 21617934 TI - The use of gentamicin-coated nails in the tibia: preliminary results of a prospective study. AB - BACKGROUND: The use of antibiotic-coated implants may reduce the rate of infection and facilitate fracture healing after surgical treatment of tibial shaft fractures. A new biodegradable gentamicin-loaded coating of an implant (UTN PROtect) was CE-certified in August 2005. In this prospective, non-randomized case series, we investigated the clinical, laboratory and radiological outcomes of 21 patients who underwent surgical treatment in closed or open tibial fractures, as well as revisions with the UTN PROtect gentamicin-coated intramedullary nail. METHODS: Of 21 patients (13 men, 8 women), 19 completed the 6-month follow-up. The study population included patients with complex tibial fractures and late revision cases. Clinical outcomes comprised adverse events, including infections and the SF-36 physical score. Laboratory outcomes, including C-reactive protein and leukocyte count as inflammatory markers, haemoglobin and serum gentamicin, were measured at baseline and up to 6 months post operatively. Radiographic assessments of fracture healing and weight-bearing capacity were determined at 5 weeks, 3 and 6 months after surgery. RESULTS: No implant-related infections occurred; one patient had superficial wound healing problems. Mean C reactive protein levels remained below 5 mg/L throughout the study, with a peak at 4-7 days after surgery (4.4 mg/L; range 0.5-16.1 mg/L). Leukocyte counts and haemoglobin levels did not vary over time during the study. The mean SF-36 physical score at 6 months was 42.6 (range 19.4-56.7). Radiographic union defined as three or four bridged cortices was achieved in 11 patients (58%) after 6 months. The remaining eight patients showed partial fracture healing with one or two bridged cortices. Additionally, 13 patients (68%) demonstrated full weight bearing capacity after 6 months. CONCLUSIONS: The use of the UTN PROtect intramedullary nail was associated with good clinical, laboratory and radiological outcomes after 6 months. These preliminary results support the use of gentamicin-coated implants as a new potential treatment option for the prevention of infection in trauma patients and in revision cases. LEVEL OF EVIDENCE: Level II. PMID- 21617935 TI - Radiological prognostic factors in patients with pandemic H1N1 (pH1N1) infection requiring hospital admission. AB - The aim of this study was to determine the radiologic findings associated with admission to the intensive care unit (ICU) and the development of acute respiratory distress syndrome (ARDS) in patients with pH1N1 infection. One hundred and four patients (15-96 years) with laboratory-confirmed pH1N1 infection seen at the Emergency Department from July to December 2009 who underwent chest radiographs were studied. Radiographs were evaluated for consolidation, ground glass opacities, interstitial patterns, distribution, and extent of findings. Eighty-seven (83.7%) of the patients were managed in the ward, and 17 (16.3%) patients eventually required admission to the ICU. All patients admitted to the ICU showed abnormalities on the initial radiograph. The presence of consolidation, multifocal, diffuse, and bilateral involvement on the initial radiograph was associated with a statistically higher risk of requiring ICU admission (p<0.001). There were no significant differences regarding age, sex, and presence of underlying comorbidities. Evolution to ARDS was found in eight cases that necessitated ICU care. All of them had on the initial radiograph patchy multifocal consolidations (p<0.001) with bilateral lesions in six cases. A higher number of lung zones involved and consolidation on the initial chest radiograph as well as a rapid progression of the radiological abnormalities were identified in patients requiring ICU admission and development of ARDS. Initial chest radiographs show acute abnormalities in all patients with severe disease. The findings of a multifocal patchy consolidation pattern with bilateral or diffuse lung involvement on admission should alert of the impending severity of disease and the risk of necessitating ICU admission. PMID- 21617936 TI - Retropharyngeal calcific tendinitis--longus colli tendinitis--an unusual cause of acute dysphagia. PMID- 21617938 TI - Non-point contamination homogenizes the water quality of pampean streams. AB - The aim of this research was to study the effects on streams water quality of non point contamination as agriculture and cattle farming activities at a basin with pampean characteristics located at the upper Reconquista River Basin. Eight reaches with different uses in their neighboring zones were selected with the proposed to detect differences at the water quality among them. SRP range was 0 0.60 mgP.PO (4) (-3) /L, ammonia 1-137.3 MUgN-NH4(+)/L and nitrate 0-4.15 mgN NO(3) (-)/L. There was high similitude and homogeneous physicochemical characteristics at the different reaches of the streams. The high levels of dissolved nutrients showed similar eutrophication conditions at the streams. PMID- 21617939 TI - Behavior of beta cyfluthrin and imidacloprid in/on mango (Mangifera indica L.). AB - Residue persistence of beta cyfluthrin and imidacloprid on mango was carried out after giving spray application of the combination formulation, beta cyfluthrin 9% + imidacloprid 21% (Solomon 300 OD) 3 times at the fruit formation stage. The treatments were, untreated control, standard dose of 75 g a.i. ha(-1) and double dose of 150 g a.i. ha(-1). Initial residues of beta cyfluthrin on mango fruits were 0.04 and 0.12 mg kg(-1) from treatments at the standard and double doses, respectively. The residues dissipated with the half-life of 2.4 and 2.6 days and persisted for 5 days only. Initial residues of imidacloprid on mango fruits were 0.14 and 0.18 mg kg(-1) from treatments at the standard and double doses, respectively. Imidacloprid residues degraded with the half-life of 3.06 and 4.16 days, respectively and persisted for 10 days. Mature mango fruits at harvest were free from residues of both insecticides. A safe pre-harvest interval of 8 days is recommended for consumption of mango fruits after treatment of the combination formulation. PMID- 21617937 TI - Intestinal microbiota in human health and disease: the impact of probiotics. AB - The complex communities of microorganisms that colonise the human gastrointestinal tract play an important role in human health. The development of culture-independent molecular techniques has provided new insights in the composition and diversity of the intestinal microbiota. Here, we summarise the present state of the art on the intestinal microbiota with specific attention for the application of high-throughput functional microbiomic approaches to determine the contribution of the intestinal microbiota to human health. Moreover, we review the association between dysbiosis of the microbiota and both intestinal and extra-intestinal diseases. Finally, we discuss the potential of probiotic microorganism to modulate the intestinal microbiota and thereby contribute to health and well-being. The effects of probiotic consumption on the intestinal microbiota are addressed, as well as the development of tailor-made probiotics designed for specific aberrations that are associated with microbial dysbiosis. PMID- 21617940 TI - Combined analysis of the association between p73 G4C14-to-A4T14 polymorphisms and cancer risk. AB - P73 is a structural and functional homologue of p53, and plays an important role in regulating cell cycle and apoptosis. A potentially functional polymorphism (designated as p73 G4C14-to-A4T14) has been identified in a region in exon 2 of the p73 gene, which may theoretically form a stem-loop structure and thereby affect p73 expression. Several investigations have reported the correlation between p73 G4C14-to-A4T14 polymorphism and cancer risk. However, the results are inconclusive. To further assess the association between p73 polymorphism and cancer risk, we performed meta-analysis of the data sets obtained from 26 individual studies involving 8,148 cancer patients and 8,150 controls. The association between p73 G4C14-to-A4T14 polymorphism and cancer risk was determined by crude odd ratios (OR) with 95% CI (confidential interval). AT allele carriers were found to have a significantly increased risk of cervical cancer (AT/GC vs. GC/GC, OR = 1.63, 95% CI = 1.14-2.33; AT/AT + AT/GC vs. GC/GC, OR = 1.49, 95% CI = 1.05-2.10), colorectal cancer (AT/AT vs. AT/GC + GC/GC, OR = 1.98, 95% CI = 1.25-3.12), head and neck cancer (AT/AT + AT/GC vs. GC/GC, OR = 1.44, 95% CI = 1.06-1.96) and other cancers (AT/AT vs. GC/GC, OR = 1.78, 95% CI = 1.24-2.57; AT/AT vs. AT/GC + GC/GC, OR = 1.80, 95% CI = 1.26-2.56). In the stratified analysis of ethnicity, a significantly elevated cancer risk was found in Caucasians (AT/AT + AT/GC vs. GC/GC, OR = 1.18, 95% CI = 1.08-1.30; allele AT vs. allele GC, OR = 1.15, 95% CI = 1.06-1.24). No significant association of p73 polymorphism with the cancer risk of smoking was detected by stratified analysis by smoking status. Together, our data suggest that the p73 G4C14-to-A4T14 may be a risk factor of cancer especially in Caucasians. PMID- 21617941 TI - Bufalin enhances the anti-proliferative effect of sorafenib on human hepatocellular carcinoma cells through downregulation of ERK. AB - The purpose of this study was to investigate the effect of bufalin on the anti proliferative activity of sorafenib in the human hepatocellular carcinoma (HCC) cell lines PLC/PRF/5 and Hep G-2 and to determine the relevant molecular mechanism. Concurrent treatment with sorafenib and bufalin at a fixed ratio (25:1) for 48 h resulted in synergistic growth inhibition in HCC cell lines as determined by CCK-8 cell viability assays. Exposure of both PLC/PRF/5 and Hep G-2 cells to this combination of sorafenib (6.25 MUM) and bufalin (50 nM) resulted in noticeable increases in apoptotic cell death, as evidenced by the disruption of mitochondria, compared to treatment with either agent alone. Although both sorafenib (6.25 MUM) and bufalin (250 nM) alone inhibited the phosphorylation of ERK, the reduction in pERK was more pronounced in the cells treated with a combination of bufalin (50 nM) and sorafenib (250 nM). Furthermore, the inhibitory effect of bufalin on pERK was blocked by the PI3kinase inhibitor LY294002, suggesting that the reduction in pERK induced by bufalin might be mediated by AKT in these two HCC cell lines. Taken together, the results of our study suggest that bufalin enhances the anti-cancer effects of sorafenib on PLC/PRF/5 and Hep G-2 by contributing to the downregulation of ERK. PMID- 21617942 TI - Gene variants of XRCC4 and XRCC3 and their association with risk for urothelial bladder cancer. AB - The DNA double strand break repair gene XRCC4, an important caretaker of genome stability and XRCC3 are suggested to play an imperative role in the development of carcinogenesis. However, no evidence has been provided showing that these genes are associated with risk of urinary bladder cancer (UBC). The study was designed to examine the polymorphisms associated with two genes namely XRCC4 G1394T (rs6869366), intron 3 (rs28360317), intron 7 rs1805377 and rs2836007 and XRCC3 (rs861539 and rs1799796), respectively and investigate their role as susceptible markers for UBC risk in North Indian cohort. In this hospital-based case-control study histologically confirmed 211 UBC patients and 244 age and gender matched controls of similar ethnicity were genotyped by means of PCR-RFLP. Significant different distributions in the frequency of the XRCC4 intron 3 genotype, but not the XRCC4 G1394T or intron 7 genotypes, between the UBC and control groups were observed. XRCC4 intron 7 Del/Del conferred enhanced risk (OR 1.94; P 0.017) in UBC. Interestingly, XRCC -1394 G>T variant genotype GG was associated with reduced risk (OR 0.27; P 0.020). However, none of the four polymorphisms in XRCC4 were associated with tobacco smoking and risk of recurrence in patients treated with BCG immunotherapy. Similarly, none of the XRCC3 polymorphisms were associated with UBC susceptibility. Our results suggested that the XRCC4 intron 3 rs6869366 genotype and intron 7 rs28360317 may be associated with UBC risk and may be a novel useful marker for primary prevention and anticancer intervention. PMID- 21617943 TI - 2-hydroxylethyl methacrylate (HEMA), a tooth restoration component, exerts its genotoxic effects in human gingival fibroblasts trough methacrylic acid, an immediate product of its degradation. AB - HEMA (2-hydroxyethyl methacrylate), a methacrylate commonly used in dentistry, was reported to induce genotoxic effects, but their mechanism is not fully understood. HEMA may be degraded by the oral cavity esterases or through mechanical stress following the chewing process. Methacrylic acid (MAA) is the primary product of HEMA degradation. In the present work we compared cytotoxic and genotoxic effects induced by HEMA and MAA in human gingival fibroblasts (HGFs). A 6-h exposure to HEMA or MAA induced a weak decrease in the viability of HGFs. Neither HEMA nor MAA induced strand breaks in the isolated plasmid DNA, but both compounds evoked DNA damage in HGFs, as evaluated by the alkaline comet assay. Oxidative modifications to the DNA bases were monitored by the DNA repair enzymes Endo III and Fpg. DNA damage induced by HEMA and MAA was not persistent and was removed during a 120 min repair incubation. Results from the neutral comet assay indicated that both compounds induced DNA double strand breaks (DSBs) and they were confirmed by the gamma-H2AX assay. Both compounds induced apoptosis and perturbed the cell cycle. Therefore, methacrylic acid, a product of HEMA degradation, may be involved in its cytotoxic and genotoxic action. PMID- 21617944 TI - Arachidonic acid omega-hydroxylase CYP4A11: inter-ethnic variations in the 8590T>C loss-of-function variant. AB - The human Cytochrome P450 4A11 (CYP4A11) is a major omega-hydroxylase involved in the regulation of blood pressure in the kidney through the conversion of arachidonic acid into 20-hydroxyeicosatetraenoic acid (20-HETE). Previous studies have reported a significant association between the 8590T>C genetic variant of CYP4A11 and hypertension. Interestingly, several population-based studies have reported ethnic differences in the prevalence of hypertension, with the highest prevalence in African populations. The aim of this work was to determine the frequency and inter-ethnic comparison of the CYP4A11 (8590T>C) functional polymorphism, in five new ethnic groups: European (99 French Caucasians), African (36 Gabonese and 50 Senegalese), South American (60 Peruvians) and North African (53 Tunisians) populations, using polymerase chain reaction-single strand conformational polymorphism and sequencing strategies. We confirmed that the CYP4A11 (8590T>C) functional polymorphism exhibits inter-ethnic frequency differences. Noteworthy, the highest 8590C allele frequency was observed in the Tunisian (30.2%), followed by Senegalese (20%) populations. In addition, the CC genotype was only found in the Gabonese and Tunisian populations (5.6% and 8.4%, respectively). These populations may be of major interest to help to clarify the linkage between hypertension and CYP4A11 (8590T>C) genotype in African populations. These findings provide data for further studies that investigate the potential association of CYP4A11 (8590T>C) variant with an incidence of hypertension genesis in respect of ethnicity. PMID- 21617945 TI - Genetic variation of new 21 autosomal short tandem repeat loci in a Chinese Salar ethnic group. AB - In the present study, we reported the allele frequencies for new 21 autosomal short tandem repeat (STR) loci, including D6S474, D12ATA63, D22S1045, D10S1248, D1S1677, D11S4463, D1S1627, D3S4529, D2S441, D6S1017, D4S2408, D19S433, D17S1301, D1GATA113, D18S853, D20S482, D14S1434, D9S1122, D2S1776, D10S1435 and D5S2500 loci. Forensic statistical parameters were estimated from a sample set of 120 unrelated healthy individuals from the Salar ethnic group in Xunhua Salar Autonomous County of Qinghai province, China. A total of 151 alleles were observed at 21 STR loci in the population, and their allele frequencies were in the range of 0.004-0.554. All STR loci showed a high degree of genetic polymorphisms, and the combined probability of exclusion, combined power of discrimination and combined probability of matching for all 21 STR loci were 0.9999993134, 0.99999999999999999991739 and 8.2607 * 10(-20), respectively. For all the 21 STR loci in the Salar ethnic group, the observed genotypic data showed no significant deviation from those expected under the Hardy-Weinberg equilibrium. The allele frequency distributions for the 21 autosomal STR loci were compared between the Salar group and its neighboring populations and significant differences were detected among these populations at D1S1677, D2S441, D3S4529, D4S2408, D6S1017, D11S4463, D12ATA63, D14S14343, D18S853, D19S433 and D22S1045 loci. PMID- 21617946 TI - Association of bovine carcass phenotypes with genes in an adaptive thermogenesis pathway. AB - Associations of carcass phenotypes with genes regulating fat and energy metabolism involved in adaptive thermogenesis were examined in beef cattle. Carcass weight (CW) was found to be associated with MAP2K6 and UCP2 genes; back fat thickness (BFT) was found to be associated with PPARGC1A, MAP2K6, and UCP2 genes; marbling score (MS) was found to be associated with PPARGC1A and MAP2K6 genes; and eye-muscle area (EMA) was found to be associated only with UCP2 gene (P < 0.05). Further analyses found significant associations of interactions between PPARGC1A and MAP2K6 genes with CW and MS. Especially, interactive genetic associations were identified between c.424 and 222 G>A in PPARGC1A and c.17-10118 T>G in MAP2K6 and between c.228+28619 A>G in PPARGC1A and c.17-10118 T>G in MAP2K6, and they were both detected for CW and MS at a significant level (P < 0.05). The current study suggested that the individual and interactive associations of PPARGC1A, MAP2K6, and UCP2 genes with carcass phenotypes might be resulted from the pathway with fat and energy metabolism through the adaptive thermogenesis. PMID- 21617947 TI - A 304 bp insertion/deletion mutation in promoter region induces the increase of porcine IDH3beta gene expression. AB - Obese and lean pig breeds show obvious differences in adipose metabolism/fat deposition; however, the molecular mechanism underlying phenotype variation remains unknown. In order to understand it, we analyzed the differences of gene expression in backfat between Meishan (a typical Chinese indigenous obese breed) and Large White (a lean Western breed) pigs. Here, we cloned porcine beta subunit of IDH3 (IDH3B) and 2447 bp 5'-flanking sequence of this gene, and determined the genomic structure. Porcine IDH3B contains three isoforms, IDH3B ( 1 ), IDH3B ( 2 ) and IDH3B ( 3 ). Real-time RT-PCR revealed that these three isoforms were prevalently up-regulated in backfat of western commercial pigs, Large White, Landrace and Duroc, compared with Chinese indigenous breeds, Meishan and Tongcheng pigs. A 304 bp insertion/deletion variant was found in the 5'-flanking region. Dual-luciferase reporter assays showed that in vitro the promoter of IDH3B gene with the insertion had higher luciferase activity as compared with the wild type. Three genotypes AA, AB and BB, due to this insertion, were detected, and the frequency of allele A was dominant in western commercial pigs, whereas allele B predominated in Chinese indigenous breeds. IDH3B mRNA expression in Meishan pigs was more abundant with genotype AA than with genotype AB or BB, as in Large White pigs. In addition, the polymorphism was detected in 317 pigs of a Large White * Meishan F2 resource population. Association analysis showed that pigs with genotype AA possessed higher backfat thickness at buttocks than those with genotype AB (P < 0.05) or BB. These data suggested that the 304 bp insertion mutation in promoter region increased the expression of porcine IDH3beta transcripts and this mutation might be a candidate marker for marker assistant selection in swine breeding. PMID- 21617948 TI - Molecular cloning and functional analysis of one ZEITLUPE homolog GmZTL3 in soybean. AB - ZEITLUPE (ZTL) plays an important role in the control of flowering time and photomorpogenesis in Arabidopsis and is highly conserved throughout the plant kingdom. Here, we report the characterization of a soybean ZTL homolog GmZTL3 (Glycine max ZTL 3). The absorption spectrum of the recombinant GmZTL3 proteins indicates that it may be a UV/blue photoreceptor. The GmZTL3 expression is independent of diurnal cycles and varies in different tissues along with developmental stages. Before the unifoliolates open fully, GmZTL3 transcripts concentrate in the roots and hypocotyls, while at flowering GmZTL3 accumulates at higher abundance in stems and petioles. Furthermore, the GmZTL3 mRNA accumulates in all kinds of leaves before flowering and concentrates in maturation seeds. In Arabidopsis, the ectopic expression of GmZTL3 delays flowering, implicating GmZTL3 is an inhibitor of flowering induction. Our data indicate that GmZTL3 probably functions as a photoreceptor and plays a role in multiple developmental processes, including the control of flowering time. PMID- 21617949 TI - Role of post translational modifications and novel crosstalk between phosphorylation and O-beta-GlcNAc modifications in human claudin-1, -3 and -4. AB - The precise characterization of post translational modifications (PTMs) is important for the understanding of protein regulatory mechanisms and their role in disease. However, experimental studies on PTMs, especially with multifunctional proteins are difficult to follow and investigate. Bioinformatic tools are therefore helpful in predicting key protein modifications. To study the role of PTMs in claudin proteins, specifically claudin-1, -3 and -4 in the onset or progression of human cancers, we performed an in silico study of various PTMs and investigated their interplay. Given that the activity of claudins is known to be influenced by two types of PTMs, specifically palmitoylation and kinase- dependent phosphorylation, we predicted two conserved regions in the topological domains of claudin-1, -3 and -4 as potential palmitoylation sites. Furthermore, conserved phosphorylation residues, which may be targets for kinases and can alter claudin's ability to maintain the integrity of tight junctions, were identified. To our knowledge, this is the first report to suggest O-glycosylation of claudin proteins, as well as a potential novel interplay between phosphorylation and O-glycosylation at Yin Yang sites. Thus, our findings may facilitate the production of anti-cancer drugs, and suggest that novel therapeutic strategies should target post translational events. PMID- 21617950 TI - Atypical meningioma of sylvian fissure with a 20-year history: a rare case report. AB - Atypical meningiomas of sylvian fissure are extremely rare. We reported a case of sylvian fissure atypical meningioma with a 20-year history. The tumor was excised subtotally, thereafter a postoperative radiation therapy was done. The patient had a favorable outcome during the two-year follow-up. The possible pathogenesis of this case was hypothesized and analyzed in this article. PMID- 21617952 TI - Genetic influences on five measures of processing speed and their covariation with general cognitive ability in the elderly: the older Australian twins study. AB - Processing speed (PS) is one of the basic elements of cognitive functions and has been regarded as a "common mechanism" which mediates general cognitive decline in aging. The present study of Australian twins (117 monozygotic pairs, 98 dizygotic pairs, and 42 single twins aged 65 years and over), estimated the genetic influences in five measures of PS: Digit Symbol Coding (DS), Trail Making Test A (TMTA), Stroop color naming and word reading (Stroop), Simple Reaction Time (SRT) and Complex Reaction Time (CRT); and their covariation with general cognitive ability (GCA): reasoning, problem-solving, and memory. Additive genetic factors explained 62% of the variance in DS, 42% in TMTA, 57% in Stroop, and 48% and 35% in SRT and CRT, respectively. Quantitative genetic modeling showed that all of the covariation between the five PS measures and GCA could be explained by one common genetic factor, while the covariation between the PS measures was partly explained by non-shared environmental as well as genetic influences. The genetic correlation among the PS measures was strongest for DS and TMTA, and between the PS measures and GCA was strongest for DS. These findings suggest that the different PS measures, as well as GCA were to a large extent influenced by the same set of genes and that the relationship between PS and GCA is entirely due to shared-genetic influences. PMID- 21617951 TI - Expression of Bcl-2 and Bax after hippocampal ischemia in DHA + EPA treated rats. AB - To determine the impact of omega3 fatty acids on post-ischemic expression of pro- and anti-apoptotic proteins in hippocampus, male rats were received 10 or 100 mg/kg [Docosahexaenoic acid (DHA) + Ecosapentaenoic acid (EPA); gavage; 21 days before ischemia to 2-10 days after ischemia]. Global cerebral ischemia reperfusion (IR) was performed using the four-vessel occlusion model; ischemia 8 min and reperfusion 6, 48 h and 10 days. IR increased Bcl-2 and Bax expression after 48 h (p < 0.05 and p < 0.01 vs. sham) and 10 days (only Bax; p < 0.05), without significant difference with DHA + EPA groups after 6 h. But after 48 h expression of Bcl-2 increased (p < 0.05 vs. IR) and Bax decreased (p < 0.05). At day 10 after ischemia expression of Bax in DHA + EPA acid groups was less than IR (p < 0.05) and in 100 mg/kg DHA + EPA group Bcl-2 expression was more than IR (p < 0.05). These data suggested that long-term gavage with DHA + EPA increase hippocampal neurons survival for days after ischemia, revealed by increased Bcl-2 and decreased Bax expressions. PMID- 21617953 TI - Prior mating experience modulates the dispersal of Drosophila in males more than in females. AB - Cues from both an animal's internal physiological state and its local environment may influence its decision to disperse. However, identifying and quantifying the causative factors underlying the initiation of dispersal is difficult in uncontrolled natural settings. In this study, we automatically monitored the movement of fruit flies and examined the influence of food availability, sex, and reproductive status on their dispersal between laboratory environments. In general, flies with mating experience behave as if they are hungrier than virgin flies, leaving at a greater rate when food is unavailable and staying longer when it is available. Males dispersed at a higher rate and were more active than females when food was unavailable, but tended to stay longer in environments containing food than did females. We found no significant relationship between weight and activity, suggesting the behavioral differences between males and females are caused by an intrinsic factor relating to the sex of a fly and not simply its body size. Finally, we observed a significant difference between the dispersal of the natural isolate used throughout this study and the widely-used laboratory strain, Canton-S, and show that the difference cannot be explained by allelic differences in the foraging gene. PMID- 21617954 TI - Coronary-cameral fistula from the sinoatrial nodal branch of the right coronary artery draining into the right atrium: demonstration by multidetector CT. PMID- 21617955 TI - Spontaneous intracranial hypotension due to intradural thoracic osteophyte with superimposed disc herniation: report of two cases. AB - Spontaneous intracranial hypotension (SIH) is a clinical syndrome in which absolute or relative hypovolemia of the cerebrospinal fluid (CSF) results in various neurological symptoms. The etiology of spontaneous CSF leaks often remains unknown. However, it is believed that the most common cause is the fragility of spinal meninges at the level of radicular nerve root sleeve. These tears can be spontaneous (primary) or secondary. Spinal pathologies can cause this tear with resultant CSF leak and SIH, which include spinal trauma, degenerative diseases and spinal surgery. Uncommonly, SIH is developed by osteophyte with disc herniation without any other pathology. In this article, we reported two cases of SIH secondary to spinal dural tear due to intradural thoracic osteophyte with superimposed disc herniation, with the absence of other pathologies, which were treated successfully with epidural blood patch (EBP). PMID- 21617956 TI - Endoscopic excision of a ganglion cyst in an infrapatellar fat pad extending into the subcutaneous layer. PMID- 21617957 TI - Acute hydrops in a host cornea after penetrating keratoplasty for keratoconus. PMID- 21617958 TI - Suprachoroidal implant surgery in intractable glaucoma. AB - PURPOSE: To report the early results of suprachoroidal silicone implant surgery in intractable glaucoma. MATERIALS AND METHODS: A modified silicone implant with no valve was implanted into the suprachoroidal space of 15 eyes with intractable glaucoma. RESULTS: The mean age of the patients was 53.0 +/- 24.5 (range 7-85) years, the mean follow-up time was 17.1 +/- 4.8 (range 10-28) months, and the mean preoperative intraocular pressure (IOP) of patients receiving two or more medications was 33.1 +/- 9.8 mmHg. At the last follow-up visit, mean IOP was 16.5 +/- 7.9 (range 10-35) mmHg (Wilcoxon signed rank test, p = 0.001). The functional success, i.e., IOP <=21 mmHg both with and without antiglaucomatous drugs, was 93.3%. The total success rate, i.e., IOP <=21 mmHg without medication, was 13.3%. The average number of antiglaucomatous drugs used was 3.8 (range 2-5) preoperatively, and 2.2 postoperatively (range 0-4) (Wilcoxon signed rank test, p = 0.011). There was a >=30% decrease in the IOP of 66.6% of the eyes. Shallow choroidal detachment as proof of drainage was evident in all cases. CONCLUSION: Drainage of the aqueous humor from the anterior chamber to the suprachoroidal space via implantation of a modified silicone implant is effective in lowering the IOP in intractable glaucoma. PMID- 21617959 TI - Macular detachment after successful intravitreal bevacizumab for myopic choroidal neovascularization. AB - PURPOSE: To determine the incidence and features of macular retinal detachment (RD) after successful intravitreal injection of bevacizumab (IVB) for the treatment of myopic choroidal neovascularizations (CNVs). METHODS: Seventy-four eyes of 74 patients who received IVB for myopic CNV were studied. Fluorescein angiography (FA) and optical coherence tomography (OCT) were performed at baseline, at 1 week after IVB, and every month thereafter. The shortest follow-up period was 6 months. RESULTS: Four eyes (5.4%) either developed or had increased macular RD after successful IVB. In the three eyes, macular RD newly developed 1 month after IVB despite the absence of dye leakage. The remaining eye had a limited area of macular RD at baseline and increased despite the disappearance of dye leakage from the CNV after the IVB. The four eyes that developed a macular RD had retinoschisis around the CNV before IVB, whereas none of the 70 other patients had a retinoschisis throughout the follow-up period. In all four eyes that either developed or had increased macular RD, outer lamellar holes were detected at the upper edge of the RD by OCT examinations, whereas none of the patients who did not develop a macular RD after IVB showed outer lamellar holes. CONCLUSIONS: These results suggest that there is a risk of developing macular RD after IVB in eyes with myopic CNV. Periodic OCT examinations are recommended for patients with myopic CNV to search for RDs after IVB. PMID- 21617961 TI - Atypical presentation of acute macular neuroretinopathy with tiny parafoveal reddish-brown lesions. AB - PURPOSE: To report two cases of acute macular neuroretinopathy with unusually small parafoveal lesions, which showed later recurrent lesions either in the same eye or the fellow eye. METHODS: Observational case series. PATIENTS: In case 1, a 48-year-old woman developed a sudden onset of a tiny paracentral scotoma associated with a small reddish-brown paracentral lesion. A new paracentral lesion with a corresponding scotoma developed 1 year later. In case 2, a 39-year old man with diabetes type 1 developed a paracentral scotoma OS with a corresponding small reddish-brown lesion. Two months later, a similar lesion with a corresponding scotoma developed in the fellow eye. CONCLUSIONS: Acute macular neuroretinopathy may present with tiny paracentral lesions difficult to detect with fundus examination, and may be associated with recurrent lesions either in the same eye or the fellow eye. PMID- 21617960 TI - Expression of interleukin-4 receptor alpha in human corneal epithelial cells. AB - PURPOSE: We previously reported that human conjunctival epithelial cells expressed functioning interleukin-4 receptor alpha (IL-4Ralpha). In this study, we investigated whether human corneal epithelial cells also express functioning IL-4Ralpha. METHODS: The presence of IL-4Ralpha mRNA and protein in human corneal epithelium was examined by reverse-transcriptase polymerase chain reaction (RT PCR) and immunohistology, respectively. The cell surface expression of IL-4Ralpha and the transcripts upregulated upon IL-4Ralpha ligand (IL-4 or IL-13) stimulation were examined by flow cytometry and quantitative RT-PCR, respectively, using immortalized human corneal-limbal epithelial (HCLE) cells. RESULTS: The mRNA and protein of IL-4Ralpha were detected in human corneal epithelium. Flow cytometry analysis showed the cell surface expression of IL 4Ralpha protein. Quantitative RT-PCR assay of HCLE cells showed the upregulation of the transcripts tumor necrosis factor alpha-induced protein 6 (TNFAIP6), RAS guanyl-releasing protein 1 (RASGRP1), carbonic anhydrase II (CA2), cytokine inducible SH2-containing protein (CISH), hyaluronan synthase 3 (HAS3), calpain 14 (CAPN14), endothelin receptor type A (EDNRA), cathepsin C (CTSC), and lecithin retinol acyltransferase (LRAT) as well as human conjunctival epithelial cells. CONCLUSION: Human corneal epithelial cells expressed functioning IL-4Ralpha, and stimulation of its ligands, IL-4 and IL-13, could induce the expression of various genes, e.g., antiinflammatory molecule genes such as TNFAIP6 and CISH and cellular differentiation and proliferation-related molecule genes such as RASGRP1, HAS3, EDNRA, and LRAT. PMID- 21617962 TI - The role of progestogens in regulating matrix metalloproteinase activity in macrophages and microglial cells. AB - Although the systemic effects of progestogens have been extensively studied, little is known in regards to the cellular effects of these compounds. Using a cellular model for vascular (macrophages) and brain (microglial) cells, we studied the effects of various progestogens, either alone or in combination with 17beta-estradiol (E(2)) on the activity of matrix metalloproteinase-9 (MMP-9), a proteolytic enzyme involved in vascular remodeling and plaque destabilization in cardiovascular events, blood-brain barrier breakdown in stroke and brain regeneration and neurovascular remodeling during repair phases of brain injury. In the absence of E(2), medroxyprogesterone acetate (MPA), a synthetic progestogen and progesterone (PG) metabolites tended to increase MMP-9 enzyme activity in macrophages and microglial cells, whereas PG decreased such activity in macrophages; exceptions being that MPA and the PG metabolite, pregnanediol (Pdiol) had no effect on macrophage MMP-9 enzyme activity and PG had no effect on microglial cell MMP-9 enzyme activity. In the presence of E(2), an opposite affect was observed whereby MPA and the PG metabolites tended to decrease MMP-9 enzyme activity from macrophages and microglial cells, whereas PG had no effect; exceptions being that MPA and Pdiol had no effect on macrophage MMP-9 enzyme activity. In conclusion, these results demonstrate that the effects of PG, PG metabolites and MPA on MMP-9 enzyme activity differ across vascular and brain cells when administered alone or in combination with E(2) which could have important mechanistic implications for hormone therapy. PMID- 21617963 TI - Identification of specific pluripotent stem cell death--inducing small molecules by chemical screening. AB - A potential application of embryonic and inducible pluripotent stem cells for the therapy of degenerative diseases involves pure somatic cells, free of tumorigenic undifferentiated embryonic and inducible pluripotent stem cells. In complex collections of chemicals with pharmacological potential we expect to find molecules able to induce specific pluripotent stem cell death, which could be used in some cell therapy settings to eliminate undifferentiated cells. Therefore, we have screened a chemical library of 1120 small chemicals to identify compounds that induce specifically apoptotic cell death in undifferentiated mouse embryonic stem cells (ESCs). Interestingly, three compounds currently used as clinically approved drugs, nortriptyline, benzethonium chloride and methylbenzethonium chloride, induced differential effects in cell viability in ESCs versus mouse embryonic fibroblasts (MEFs). Nortriptyline induced apoptotic cell death in MEFs but not in ESCs, whereas benzethonium and methylbenzethonium chloride showed the opposite effect. Nortriptyline, a tricyclic antidepressant, has also been described as a potent inhibitor of mitochondrial permeability transition, one of two major mechanisms involved in mitochondrial membrane permeabilization during apoptosis. Benzethonium chloride and methylbenzethonium chloride are quaternary ammonium salts used as antimicrobial agents with broad spectrum and have also been described as anticancer agents. A similar effect of benzethonium chloride was observed in human induced pluripotent stem cells (hiPSCs) when compared to both primary human skin fibroblasts and an established human fibroblast cell line. Human fibroblasts and hiPSCs were similarly resistant to nortriptyline, although with a different behavior. Our results indicate differential sensitivity of ESCs, hiPSCs and fibroblasts to certain chemical compounds, which might have important applications in the stem cell-based therapy by eliminating undifferentiated pluripotent stem cells from stem cell-derived somatic cells to prevent tumor formation after transplantation for therapy of degenerative diseases. PMID- 21617964 TI - Determination of noise induced hearing loss in mining: an application of hierarchical loglinear modelling. AB - The levels of noise arise from mining industry seem to be higher when compared to other industries. For this reason, noise exposure and noise-induced hearing loss (NIHL) are prevalent in mining. Assessment of noise emission levels that arise from various mining operations is required to prevent and minimize the NIHL. Because the studies for preventing occupational hearing loss among miners are inadequate, a quarry and stone crushing-screening plant was selected to generate site-specific data. The noise levels of the environments in which workers work were measured and also a hearing test centre applied hearing tests to the workers. According to the hearing test results, it was determined that the part of workers have hearing loss. The main factors affecting the NIHL were assumed as experience, noise level, miners' age and occupation, and by taking into account the sub factors of the main factors, multi way contingency tables were prepared. Then hierarchical loglinear analysis method was implemented to categorized data; thus, the probabilities might effect NIHL was investigated. At the end of this study, it was found that the most risky occupation group was the drivers, and additionally, these workers were mostly exposed to 70-79 dB(A) noise level. When the important interactions are evaluated, it is determined that 4-11 years experienced crusher workers have high probability of NIHL because of high exposure to 90-99 dB(A) noise level. Moreover, the most important interactions which may affect the NIHL were identified and the precautions to reduce hearing loss were presented. PMID- 21617965 TI - Major ion chemistry of shallow groundwater of a fast growing city of central India. AB - Nagpur City located in semiarid area of central India is a fast-growing industrial centre. In recent years, rapid development has created an increased demand for drinking water, which is increasingly being fulfilled by groundwater abstraction. The present study was undertaken to assess major ion chemistry of shallow groundwater to understand geochemical evolution of groundwater and water quality for promoting sustainable development and effective management of groundwater resources. A total of 47 water samples were collected from shallow aquifer of selected parts of the city and the water chemistry of various ions viz. Ca(2 +), Mg(2 +), Na(+), K(+), CO(3)(2-), HCO(3)(-), Cl(-), SO(4)(2-) and NO(3)(-) are carried out. The chemical relationships in Piper diagram identify Ca HCO(3)-Cl and mixed Ca-Na-HCO(3)-Cl as most prevalent water types. Alkaline earth exceeds alkalis and weak acids exceed strong acids. Ionic ratios and Gibb's diagram suggest that silicate rock weathering and anthropogenic activities are the main processes that determine the ionic composition in the study area. The nitrate appeared as a major problem of safe drinking water in this region. We recorded highest nitrate concentration, i.e., 411 mg/l in one of the dug well. A comparison of groundwater quality in relation to drinking water quality standards revealed that about half of the shallow aquifer samples are not suitable for drinking. PMID- 21617966 TI - Distribution and health risk assessment of HCHs in urban soils of Beijing, China. AB - The concentrations of hexachlorocyclohexanes (HCHs) were investigated in urban soil samples collected from business area, classical garden (CL), culture and educational area, large public green space (LA), residential area, and roadside area in Beijing. HCH concentrations ranged from 0.32 to 136.43 ng/g, with a geometric mean of 3.46 ng/g. The HCH concentrations in CL and LA were much higher than that in the other types of land use, which was due to the usage of HCHs to protect vegetation in CL and LA. Source identification showed that contamination source of HCHs was derived from historical HCHs (including technical HCHs and Lindane) as well as the long-range atmospheric transportation of HCHs. HCH concentrations showed a decreasing trend from the city centre to the suburb, and it increased with the age of the urban area. HCHs were negatively correlated with pH and positively correlated with total organic carbon and black carbon in soils. Health risk assessment with CalTOX and Monte Carlo analysis showed that health risks mainly came from dermal uptake and inhalation exposure pathways, and the total risk values were lower than the acceptable health risk value (10(- 6)). The sensitivity analysis indicated that the reaction half-life of HCHs in soil, fraction dermal uptake from soil, exposure duration, and organic carbon fraction in soil significantly contributed to the variance of the health risk. PMID- 21617967 TI - Assessing reliable indicators to sewage pollution in coastal soft-bottom communities. AB - Physicochemical characteristics of sediment and benthic communities were studied in the proximity of seven sewage outfalls with differences in flow and wastewater treatment in the Western Mediterranean Sea. Redox potential was the only abiotic parameter which showed a pattern related with distance to outfalls, whereas granulometry, percentage of organic matter, metal concentrations or pH did not show changes related with outfall presence. Benthic community analysis proved to be the most suitable monitoring tool. The results showed that the highest impacted stations corresponded with those closest to outfall with the highest flow and only pre-treatment, whilst a decrease of this tendency was detected in the locations where secondary treatment takes place. Meta-analysis showed a decrease of amphipods and tanaids abundance as well as redox potential, as the indicators with the clearest response to sewage presence. PMID- 21617968 TI - Management of special conditions in patients on vitamin K antagonists. AB - Physicians are occasionally faced with difficult situations in the management of vitamin K antagonists (VKA) due to the lack of sound data available in controlled studies on certain conditions. In this review we would like to address some special but frequent conditions that can be encountered in daily clinical practice. These include the use of VKA in hemodialysis, thromboembolism in patients with liver cirrhosis and the thromboembolic risk in patients who bleed in the course of treatment with VKA. Moreover, two other conditions were examined: what the best way of expressing prothrombin time would be in patients with liver disease and how to behave when a patient treated with VKA shows a subtherapeutic INR. These topics were discussed by a panel of experts during a workshop recently held in Milan by the Italian Federation of Centres for the Diagnosis of Thrombosis and the Surveillance of Antithrombotic Therapies (FCSA). The main aim of the workshop was to provide helpful and practical advice to physicians in the daily management of VKA. PMID- 21617970 TI - Seizure, syncope, or both: cardiac pacemaker reveals temporal lobe epilepsy. PMID- 21617969 TI - Effect of dietary arginine supplementation on reproductive performance of mice with porcine circovirus type 2 infection. AB - The objective of this study was to investigate whether supplemental dietary arginine increases reproductive performance in mice infected with porcine circovirus type2 (PCV2). A total of 50KM female mice were allotted randomly to the arginine group (0.6% arginine+gestation diet) and control group (1.22% alanine+gestation diet). All the mice began to mate after 14 days of treatment with our prepared feed and challenged with PCV2 at the dose of 100 TCID50 (50% tissue culture infection dose, TCID50) after 7 days of pregnancy. Abortion rate, litter number, litter birth weight, the daily weight gain in the first 7 days and survival rate in the first 2 weeks of the neonates were calculated. The serum progesterone, estrogen, nitric oxide and superoxide dismutase (SOD) activity and total antioxidant capacity (T-AOC) on the 14th day of pregnancy were measured. Arginine supplementation decreased the abortion rate of pregnant mice and mortality of neonates caused by PCV2 infection. Further, litter number, litter birth weight and the daily weight gain of neonates increased in the arginine group compared to the control group. Arginine supplementation increased significantly the serum progesterone (P<0.01) and nitric oxide levels (P<0.05), but had little effect on the serum estrogen level. SOD activity and T-AOC in the arginine group were significantly higher (P<0.01) than the control group. In conclusion, arginine supplementation partially reversed the reproductive failure in mice caused by PCV2 infection. PMID- 21617971 TI - Research progress on Livin protein: an inhibitor of apoptosis. AB - Livin is a member of the inhibitors of apoptosis (IAP) family, which plays crucial roles in apoptosis, cell proliferation, and cell cycle control. Abnormal Livin expression is sometimes detected during the process of cancer formation and/or progression. Thus, Livin research may provide an opportunity for the development of potential therapy for Livin-relevant cancers. In this review, we introduce Livin structure, biological function, and its role in cancer formation and progression and the possible interventions for cancer therapy. PMID- 21617972 TI - Histomorphometric analysis of inflammatory response and necrosis in re-implanted central incisor of rats treated with low-level laser therapy. AB - Low-level laser therapy is a tool employed in the management of post-operative inflammation process and in the enhancement of reparative process. The aim of the study was to perform histological evaluation of dental and periodontal ligament of rats central upper-left incisor teeth re-implanted and irradiated with low level laser (InGaAl, 685 nm, 50 J/cm(2)) 15, 30, and 60 days after re implantation. Seventy-two male rats had the central upper left incisor removed and kept for 15 min on dry gauze before replantation. Laser was irradiated over the root surface and empty alveolus prior replantation and over surrounding mucosa after the re-implantation. After histological procedures, all slices were analyzed regarding external resorption area and histological aspects. We observed an increase of root resorption (p < 0.05) in the control group compared to the laser group at 15, 30, and 60 days. These results showed that the laser groups developed less root resorption areas than the control group in all experimental periods. Additionally, histological analysis revealed less inflammatory cells and necrotic areas in laser groups. PMID- 21617973 TI - Efficacy of low-level laser therapy in the management of pain, facial swelling, and postoperative trismus after a lower third molar extraction. A preliminary study. AB - Pain, swelling, and trismus are the most common complications after surgical removal of impacted lower third molars. The aim of this study was to evaluate the analgesic and anti-inflammatory effects of a low-level laser therapy (Laser SmileTM, Biolase(r), San Clemente, USA) applied to the wound appeared after the surgical removal of impacted lower third molars. A prospective, randomized, and double-blind study was undertaken in 20 healthy patients with two symmetrically impacted lower third molars. The application of a low-level laser was made randomly on one of the two sides after surgery. The experimental side received 5 J/cm(2) of energy density, a wavelength of 810 nm, and an output power of 0.5 W. On the control side, a handpiece was applied intraorally, but the laser was not activated. Evaluations of postoperative pain, trismus, and swelling were made. The sample consisted of 11 women and nine men, and mean age was 23.35 years (18 37). The pain level in the first hours after surgery was lower in the experimental side than in the placebo side, although without statistically significant differences (p = 0.258). Swelling and trismus at the 2nd and 7th postoperative days were slightly higher in the control side, although not statistically significant differences were detected (p > 0.05). The application of a low-level laser with the parameters used in this study did not show beneficial affects in reducing pain, swelling, and trismus after removal of impacted lower third molars. PMID- 21617974 TI - One-step radiosynthesis of 18F-AlF-NOTA-RGD2 for tumor angiogenesis PET imaging. AB - PURPOSE: One of the major obstacles of the clinical translation of (18)F-labeled arginine-glycine-aspartic acid (RGD) peptides has been the laborious multistep radiosynthesis. In order to facilitate the application of RGD-based positron emission tomography (PET) probes in the clinical setting we investigated in this study the feasibility of using the chelation reaction between Al(18)F and a macrocyclic chelator-conjugated dimeric RGD peptide as a simple one-step (18)F labeling strategy for development of a PET probe for tumor angiogenesis imaging. METHODS: Dimeric cyclic peptide E[c(RGDyK)](2) (RGD(2)) was first conjugated with a macrocyclic chelator, 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA), and the resulting bioconjugate NOTA-RGD(2) was then radiofluorinated via Al(18)F intermediate to synthesize (18)F-AlF-NOTA-RGD(2). Integrin binding affinities of the peptides were assessed by a U87MG cell-based receptor binding assay using (125)I-echistatin as the radioligand. The tumor targeting efficacy and in vivo profile of (18)F-AlF-NOTA-RGD(2) were further evaluated in a subcutaneous U87MG glioblastoma xenograft model by microPET and biodistribution. RESULTS: NOTA RGD(2) was successfully (18)F-fluorinated with good yield within 40 min using the Al(18)F intermediate. The IC(50) of (19)F-AlF-NOTA-RGD(2) was determined to be 46 +/- 4.4 nM. Quantitative microPET studies demonstrated that (18)F-AlF-NOTA-RGD(2) showed high tumor uptake, fast clearance from the body, and good tumor to normal organ ratios. CONCLUSION: NOTA-RGD(2) bioconjugate has been successfully prepared and labeled with Al(18)F in one single step of radiosynthesis. The favorable in vivo performance and the short radiosynthetic route of (18)F-AlF-NOTA-RGD(2) warrant further optimization of the probe and the radiofluorination strategy to accelerate the clinical translation of (18)F-labeled RGD peptides. PMID- 21617975 TI - Evaluation of a cumulative SUV-volume histogram method for parameterizing heterogeneous intratumoural FDG uptake in non-small cell lung cancer PET studies. AB - PURPOSE: Standardized uptake values (SUV) are commonly used for quantification of whole-body [(18)F]fluoro-2-deoxy-D: -glucose (FDG) positron emission tomography (PET) studies. Changes in SUV following therapy, however, only provide a proper measure of response in case of homogeneous FDG uptake in the tumour. The purpose of this study was therefore to implement and characterize a method that enables quantification of heterogeneity in tumour FDG uptake. METHODS: Cumulative SUV volume histograms (CSH), describing % of total tumour volume above % threshold of maximum SUV (SUV(max)), were calculated. The area under a CSH curve (AUC) is a quantitative index of tumour uptake heterogeneity, with lower AUC corresponding to higher degrees of heterogeneity. Simulations of homogeneous and heterogeneous responses were performed to assess the value of AUC-CSH for measuring uptake and/or response heterogeneity. In addition, partial volume correction and image denoising was applied prior to calculating AUC-CSH. Finally, the method was applied to a number of human FDG scans. RESULTS: Partial volume correction and noise reduction improved CSH curves. Both simulations and clinical examples showed that AUC-CSH values corresponded with level of tumour heterogeneity and/or heterogeneity in response. In contrast, this correspondence was not seen with SUV(max) alone. The results indicate that the main advantage of AUC-CSH above other measures, such as 1/COV (coefficient of variation), is the possibility to measure or normalize AUC-CSH in different ways. CONCLUSION: AUC-CSH might be used as a quantitative index of heterogeneity in tracer uptake. In response monitoring studies it can be used to address heterogeneity in response. PMID- 21617976 TI - 123I-MIBG scintigraphy/SPECT versus 18F-FDG PET in paediatric neuroblastoma. AB - PURPOSE: To analyse different uptake patterns in (123)I-MIBG scintigraphy/SPECT imaging and (18)F-FDG PET in paediatric neuroblastoma patients. METHODS: We compared 23 (123)I-MIBG scintigraphy scans and 23 (18)F-FDG PET scans (mean interval 10 days) in 19 patients with a suspected neuroblastic tumour (16 neuroblastoma, 1 ganglioneuroblastoma, 1 ganglioneuroma and 1 opsomyoclonus syndrome). SPECT images of the abdomen or other tumour-affected regions were available in all patients. Indications for (18)F-FDG PET were a (123)I-MIBG negative tumour, a discrepancy in (123)I-MIBG uptake compared to the morphological imaging or imaging results inconsistent with clinical findings. A lesion was found by (123)I-MIBG scintigraphy and/or (18)F-FDG PET and/or morphological imaging. RESULTS: A total of 58 suspicious lesions (mean lesion diameter 3.8 cm) were evaluated and 18 were confirmed by histology and 40 by clinical follow-up. The sensitivities of (123)I-MIBG scintigraphy and (18)F-FDG PET were 50% and 78% and the specificities were 75% and 92%, respectively. False positive results (three (123)I-MIBG scintigraphy, one (18)F-FDG PET) were due to physiological uptake or posttherapy changes. False-negative results (23 (123)I MIBG scintigraphy, 10 (18)F-FDG PET) were due to low uptake and small lesion size. Combined (123)I-MIBG scintigraphy/(18)F-FDG PET imaging showed the highest sensitivity of 85%. In 34 lesions the (123)I-MIBG scintigraphy and morphological imaging findings were discrepant. (18)F-FDG PET correctly identified 32 of the discrepant findings. Two bone/bone marrow metastases were missed by (18)F-FDG PET. CONCLUSION: (123)I-MIBG scintigraphy and (18)F-FDG PET showed noticeable differences in their uptake patterns. (18)F-FDG PET was more sensitive and specific for the detection of neuroblastoma lesions. Our findings suggest that a (18)F-FDG PET scan may be useful in the event of discrepant or inconclusive findings on (123)I-MIBG scintigraphy/SPECT and morphological imaging. PMID- 21617977 TI - Standard uptake value and metabolic tumor volume of 18F-FDG PET/CT predict short term outcome early in the course of chemoradiotherapy in advanced non-small cell lung cancer. AB - PURPOSE: The aim of this study is to investigate the role of standard uptake values (SUVs) and metabolic tumor volume (MTV) in [(18)F]fluorodeoxyglucose positron emission tomography/computed tomography (FDG PET/CT) to predict the short-term outcome of chemoradiotherapy (CRT) in patients with advanced non-small cell lung cancer (NSCLC). METHODS: A total of 37 patients were included in the prospective study. All patients were evaluated by FDG PET before and following 40 Gy radiotherapy (RT) with a concurrent cisplatin-based chemotherapy regimen. Semiquantitative assessment was used to determine maximum and mean SUVs (SUV(max)/SUV(mean)) and metabolic tumor volume (MTV). Short-term outcome using the treatment response evaluation was assessed according to the Response Evaluation Criteria in Solid Tumors. The receiver-operating characteristic (ROC) curve analysis was used to determine the diagnostic accuracy of (18)F-FDG PET in identifying responders. RESULTS: Changes in SUV(max), SUV(mean), and MTV were significantly more pronounced in responders than in nonresponders (p = 0.002, 0.002, 0.000). The thresholds of SUV(max), SUV(mean), and MTV changes defined by ROC curve analysis were 37.2, 41.7, and 29.7%, respectively. The sensitivity, specificity, and accuracy of SUV(max) change for predicting tumor response were 83.3, 84.6, and 84.9%, respectively. The sensitivity, specificity, and accuracy of SUV(mean) change for predicting tumor response were 79.2, 100, and 88.8%, respectively. The sensitivity, specificity, and accuracy of MTV change for predicting tumor response were 91.7, 84.6, and 92.3%, respectively. CONCLUSION: SUV and MTV changes from two serial (18)F-FDG PET/CT scans, before and after initial CRT, allow prediction of the treatment response in advanced NSCLC. PMID- 21617978 TI - Focus on MALDI imaging. PMID- 21617979 TI - Profiling of non-esterified fatty acids in human plasma using liquid chromatography-electron ionization mass spectrometry. AB - This paper focuses on the development of a novel approach to analyze underivatized fatty acids in human plasma. The method is based on liquid-liquid extraction followed by reversed phase liquid chromatography coupled to direct electron ionization mass spectrometry (LC-Direct-EI-MS). The assay is validated. Calibrations show satisfactory linearity and precision in the investigated range of linearity. Recoveries span from 75% to 104%. The method limits of detection, varying from 0.53 to 5.35 MUM, are satisfactory for the quantitation of non esterified fatty acids (NEFAs) in plasma at physiological levels. The method has been successfully applied to the NEFAs profiling of plasma samples from healthy adult volunteers and subjects affected by diabetes mellitus. Compared with published protocols based on gas chromatography-mass spectrometry and liquid chromatography coupled to electrospray ionization mass spectrometry, this method does not require derivatization and does not show matrix effects, thus simplifying sample preparation procedure and reducing the total time of analysis to approximately 90 min. In addition, Direct-EI-MS allows the acquisition of high quality NIST library-matchable EI spectra, allowing an easy-to-obtain identification of the target NEFAs. PMID- 21617981 TI - Urethral closure pressures among primiparous women with and without levator ani muscle defects. AB - INTRODUCTION AND HYPOTHESIS: Vaginal birth is an established risk factor for levator ani (LA) defects and incontinence. We hypothesized an association between urethral pressure profiles and LA defects. METHODS: One hundred sixty primiparous women, 9-12 months postpartum, were assessed with MRI for LA defects, urodynamic testing, and instrumented speculum for vaginal closure force. Urodynamic testing included resting maximal urethral closure pressure (MUCP) and urethral closure pressure with a pelvic floor contraction or Kegel (KUCP). We examined the relationships between MUCP, KUCP, LA defect status, and vaginal closure force. RESULTS: There was no significant association between MUCP or KUCP in women with and without LA defects (p = 0.94, p = 0.95). Additionally, there was no correlation between MUCP and vaginal closure force (r = 0.06, p = 0.41), and a weak correlation between KUCP and vaginal closure force (r = 0.20, p = 0.01). CONCLUSIONS: In this population, urethral pressure profiles are unrelated to LA defect status after vaginal birth, indicating that the mechanism responsible for LA damage spares the urethra. PMID- 21617982 TI - Mullerianosis involving the urinary bladder. AB - We report a case of mullerianosis involving the urinary bladder. A 45-year-old woman presented with complaints of chronic pelvic pain, dysmenorrhoea, irritative lower urinary tract symptoms and cyclical haematuria. Cystoscopy and transurethral biopsy suggested mullerianosis involving the posterior bladder wall; she subsequently had surgery including partial cystectomy with complete excision of the lesion. Histology reported presence of endometrial, endocervical and endosalpingeal tissues within the bladder mass confirming mullerianosis. She made satisfactory postoperative recovery. Awareness of this lesion is necessary for proper diagnosis and appropriate treatment. PMID- 21617983 TI - Efficacy and safety of combined therapy with tamsulosin and tolterodine in female patients with a maximal flow rate less than 12 ml/s. AB - INTRODUCTION AND HYPOTHESIS: We assessed the effect of tamsulosin HCl (0.2 mg) with or without tolterodine extended release (2 mg) on female patients with a maximal flow rate (Qmax) less than 12 ml/s who were suspected of having functional bladder outlet obstruction. METHODS: From January 2007 to December 2008, 250 patients with a Qmax less than 12 ml/s were selected for this study. Initial drop-out rates in groups I (15.2%) and II (40.0%) are significantly different: 19 of 125 patients in groups I and 50 of 125 patients in group II failed to complete the 12-week clinical trial. The patients were treated with tamsulosin alone (0.2 mg/day; group I, n = 106) or with tamsulosin combined with tolterodine (2 mg/day; group II, n = 75). The effectiveness of these medications was assessed at baseline and after 12 weeks of treatment on the basis of the International Prostate Symptom Score (IPSS) and other measures including the Qmax and the postvoid residual urine volume. RESULTS: The total IPSS, the voiding symptom score, the Qmax, and the residual urine volume were significantly improved from baseline after 12 weeks of treatment (p < 0.05) in both groups, whereas the storage symptom score significantly improved only in group II (p < 0.05). After 12 weeks of treatment, there were no significant differences in subjective symptom scores or objective uroflowmetric parameters between the two groups, except for storage symptoms (group I, 4.3 +/- 1.6 vs group II, 3.8 +/- 0.9) and postvoid residual urine (group I, 31.8 +/- 22.4 vs group II, 56.1 +/- 29.7), which was not considered to be clinically meaningful. CONCLUSION: Combination therapy with tamsulosin and tolterodine improved the subjective symptoms and uroflowmetric measures of female patients with a maximal flow rate of less than 12 ml/s. Women with a slight degree of storage symptoms will not be benefitted by prescribing anticholinergics. PMID- 21617984 TI - Reverse-transcriptase quantitative PCR method to detect uptake of hydrogen produced from cyanobacteria by Alcaligenes hydrogenophilus, an aerobic hydrogen oxidising bacterium. AB - Hydrogen-oxidising bacteria play a key ecological role in a variety of habitats including the rhizosphere and hot springs. To investigate the possibly of interspecies hydrogen exchange between cyanobacteria and hydrogen-oxidising bacteria, we developed a sensitive and reliable reverse-transcriptase qPCR assay for up-regulation of the hupS gene in the knallgas bacterium Alcaligenes hydrogenophilus DSM2625. The assay detected up-regulation of the gene at initial hydrogen concentrations as low as 0.12 MUM. Expression of hupS also increased in the presence of hydrogen-producing cyanobacteria, both when Ah DSM2625 was directly added to a hydrogen-producing culture of the cyanobacteria, and when cultures were physically separated in a vessel that allowed gas exchange. Additional refinements and development of the sensitive assay will lead to a better understanding of hydrogen exchange in aerobic ecosystems and development of reporter strains to detect hydrogen-producing organisms. PMID- 21617985 TI - Peutz-Jeghers syndrome: a patient's view. AB - Life with Peutz-Jeghers syndrome (PJS) is complex and demanding, affecting not only the patients, but also their family members, loved ones, and clinicians. In general, clinicians and researchers have defined the syndrome. Now, in addition, people with PJS and their caregivers use the Internet for medical research and to find others with whom to share information and support. An online support group for people interested in PJS has been an important resource since 2000. This article is meant to complement the medical literature and clinician practice by sharing knowledge gained from this group and through the author's personal experience as a PJS patient. Clinicians and researchers may benefit from a better rounded view of the syndrome, a long list of questions and issues, and tips for future research and clinical practice. This is offered with the intent to ease suffering and improve future patient care. PMID- 21617987 TI - Task switching with a 2:1 cue-to-task mapping: separating cue disambiguation from task-rule retrieval. AB - An experiment is reported that investigated switching among two numerical judgment tasks with a factorial variation of the cue-to-task mapping (1:1 vs. 2:1) for each of the tasks. In addition, the precuing interval (CSI) was varied. The results suggest that with a long CSI of 1,100 ms, switching performance is almost completely determined by the task-specific conditional probability of a task switch given a cue switch. This effect probably reflects the complexity of task-rule retrieval. Without preparation (CSI = 0 ms), the complexity of cue disambiguation as a function of the number of cues across tasks seems to account for most part of the additional variance observed in this condition. The latter observation is in line with suggestions that increasing the number of cues per task from one to two introduces additional demands on the level of cue processing that reflect the transition from an isomorphic to a homomorphic mapping function. PMID- 21617986 TI - Extracorporeal shock wave therapy promotes cell proliferation and collagen synthesis of primary cultured human tenocytes. AB - PURPOSE: The aim of this study was to investigate whether the effects of extracorporeal shock wave therapy (ESWT) could affect the behavior of primary cultured human tenocytes over a 12-day period. METHODS: In this controlled laboratory study, primary human tenocytes were established from semitendinosus tendons collected from 3 patients undergoing arthroscopic anterior cruciate ligament (ACL) reconstruction. Cell viability, overall cell morphology, cell proliferation, and collagen synthesis following ESWT have been evaluated. RESULTS: ESWT significantly interferes with the overall cell morphology, by impairing dedifferentiation of the cells. Furthermore, a shock wave-mediated growth-promoting effect was measured by the MTT (tetrazolium) colorimetric assay and by the proliferation marker Ki67. Lastly, a significant increase in collagen (mainly type I) synthesis by ESWT-tenocytes compared with control cells was found. CONCLUSIONS: Shock wave treatment promoted cell growth and collagen synthesis of primary cultured human tenocytes. The clinical benefits of ESWT may be ascribed to an increased efficiency of tendon repair after injury. PMID- 21617988 TI - cGMP regulates hydrogen peroxide accumulation in calcium-dependent salt resistance pathway in Arabidopsis thaliana roots. AB - 3',5'-cyclic guanosine monophosphate (cGMP) is an important second messenger in plants. In the present study, roles of cGMP in salt resistance in Arabidopsis roots were investigated. Arabidopsis roots were sensitive to 100 mM NaCl treatment, displaying a great increase in electrolyte leakage and Na(+)/K(+) ratio and a decrease in gene expression of the plasma membrane (PM) H(+)-ATPase. However, application of exogenous 8Br-cGMP (an analog of cGMP), H(2)O(2) or CaCl(2) alleviated the NaCl-induced injury by maintaining a lower Na(+)/K(+) ratio and increasing the PM H(+)-ATPase gene expression. In addition, the inhibition of root elongation and seed germination under salt stress was removed by 8Br-cGMP. Further study indicated that 8Br-cGMP-induced higher NADPH levels for PM NADPH oxidase to generate H(2)O(2) by regulating glucose-6-phosphate dehydrogenase (G6PDH) activity. The effect of 8Br-cGMP and H(2)O(2) on ionic homeostasis was abolished when Ca(2+) was eliminated by glycol-bis-(2-amino ethyl ether)-N,N,N',N'-tetraacetic acid (EGTA, a Ca(2+) chelator) in Arabidopsis roots under salt stress. Taken together, cGMP could regulate H(2)O(2) accumulation in salt stress, and Ca(2+) was necessary in the cGMP-mediated signaling pathway. H(2)O(2), as the downstream component of cGMP signaling pathway, stimulated PM H(+)-ATPase gene expression. Thus, ion homeostasis was modulated for salt tolerance. PMID- 21617989 TI - WCI, a novel wheat chymotrypsin inhibitor: purification, primary structure, inhibitory properties and heterologous expression. AB - A novel chymotrypsin inhibitor, detected in the endosperm of Triticum aestivum, was purified and characterized with respect to the main physical-chemical properties. On the basis of its specificity, this inhibitor was named WCI (wheat chymotrypsin inhibitor). WCI is a monomeric neutral protein made up of 119 residues and molecular mass value of 12,933.40 Da. Automated sequence and mass spectrometry analyses, carried out on several samples of purified inhibitor, evidenced an intrinsic molecular heterogeneity due to the presence of the isoform [des-(Thr)WCI], accounting for about 40% of the total sample. In vitro, WCI acted as a strong inhibitor of bovine pancreatic chymotrypsin as well as of chymotryptic-like activities isolated from the midgut of two phytophagous insects, Helicoverpa armigera (Hub.) and Tenebrio molitor L., respectively. No inhibitory activities were detected against bacterial subtilisins, bovine pancreatic trypsin, porcine pancreatic elastase or human leukocyte elastase. The primary structure of WCI was significantly similar (45.7-89.1%) to those of several proteins belonging to the cereal trypsin/alpha-amylase inhibitor super family and showed the typical sequence motif of this crowed protein group. The cDNA of the inhibitor (wci-cDNA) was isolated from wheat immature caryopses and employed to obtain a recombinant product in E. coli. Experimental evidences indicated that the recombinant inhibitor was localized in the inclusion bodies from which it was recovered as soluble and partially active protein by applying an appropriate refolding procedure. WCI reactive site localization, as well as its inhibitory specificity, was investigated by molecular modeling approach. PMID- 21617990 TI - Diverse spontaneous silencing of a transgene among two Nicotiana species. AB - In plants, transgenes frequently become spontaneously silenced for unknown reasons. Typically, transgene silencing involves the generation of small interfering RNAs (siRNAs) that directly or indirectly target cognate DNA and mRNA sequences for methylation and degradation, respectively. In this report, we compared spontaneous silencing of a transgene in Nicotiana benthamiana and Nicotiana tabacum. In both species, abundant siRNAs were produced. In N. benthamiana, the self-silencing process involved mRNA degradation and dense DNA methylation of the homologous coding region. In N. tabacum, self-silencing occurred without complete mRNA degradation and with low methylation of the cognate coding region. Our data indicated that in plants, siRNA-mediated spontaneous silencing is, in addition to mRNA degradation, based on translational inhibition. Differences in the initiation and establishment of self-silencing together with marked differences in the degree of de novo DNA methylation showed that the mechanistic details of RNA silencing, although largely conserved, may vary also in genetically close plant species. PMID- 21617991 TI - Sclerostin levels during growth in children. AB - Serum sclerostin levels are associated with cortical porosity, suggesting that changes in sclerostin production during growth may play a role in defining cortical structure. INTRODUCTION: Sclerostin, produced by osteocytes, is a potent inhibitor of Wnt signaling and bone formation. While sclerostin levels increase with age in adults and are higher in men compared to women, there is currently no information on changes in circulating sclerostin levels during growth in humans. METHODS: We measured serum sclerostin levels in 6- to 21-year-old girls (n = 62) and boys (n = 56) and related these to trabecular and cortical bone microarchitectural parameters using high-resolution peripheral quantitative computed tomography and to markers of bone turnover. RESULTS: Serum sclerostin levels were higher in boys as compared to girls and declined in both sexes following the onset of puberty. There was no consistent relationship between sclerostin levels and trabecular bone parameters in either sex. However, serum sclerostin levels were inversely associated with cortical volumetric bone mineral density and cortical thickness in girls and positively associated with the cortical porosity index in both girls and boys. Bone turnover markers were positively correlated with serum sclerostin levels in both sexes. CONCLUSION: The gender difference in serum sclerostin levels appears to be established during puberty, and sclerostin levels tend to decline in late puberty in both girls and boys. Serum sclerostin levels are associated with cortical porosity, suggesting that changes in sclerostin production during growth may play a role in defining cortical structure. PMID- 21617992 TI - Partial adherence: a new perspective on health economic assessment in osteoporosis. AB - Partial adherence in osteoporosis increases the risk for fragility fracture and has considerable impact on cost-effectiveness. This review highlights a number of avenues for further research, such as improved definition of thresholds of compliance and persistence, as well as gap length, offset times, and fraction of benefit. INTRODUCTION: A number of economic models have been developed to evaluate osteoporosis therapies and support decisions regarding efficient allocation of health care resources. Adherence to treatment is seldom incorporated in these models, which may reduce their validity for decision-making since adherence is poor in real-world clinical practice. METHODS: An ad hoc working group of the European Society for Clinical and Economic Aspects of Osteoporosis and Osteoarthritis met to review key issues concerning the incorporation of partial adherence in health economic models. RESULTS: Observational data have shown that poor adherence is associated with an increase in the risk for fragility fracture. Health economic modelling indicates that full adherence is associated with more quality-adjusted life years gained than partial adherence, as well as higher treatment costs and lower fracture-related costs. Although adherence appears as an important driver of cost-effectiveness, the effect is dependent on a range of other variables, such as offset time, fraction of benefit, fracture risk, fracture efficacy, fracture-related costs, and drug cost, some of which are poorly defined. Current models used to evaluate cost effectiveness in osteoporosis may oversimplify the contributions of compliance and persistence. CONCLUSION: Partial adherence has a significant impact on cost effectiveness. Further research is required to optimise thresholds of compliance and persistence, the impact of gap length, offset times, and fraction of benefit. PMID- 21617993 TI - Teriparatide increases the maturation of circulating osteoblast precursors. AB - This study shows that teriparatide promotes the circulating osteoblast (OB) precursor degree of maturation in patients affected by postmenopausal osteoporosis. INTRODUCTION: Anabolic treatment with teriparatide has proven effective for the therapy of postmenopausal osteoporosis and significantly reduces the risk of non-vertebral fragility fractures. The aim of this study was to investigate the effect of teriparatide on circulating OB precursors. METHODS: We evaluated by flow cytometry and real-time PCR the expression of OBs typical markers in peripheral blood mononuclear cells during treatment with teriparatide plus calcium and vitamin D, raloxifene plus calcium and vitamin D or calcium and vitamin D alone at various time points. Serum bone alkaline phosphatase and osteocalcin (OC) were measured as markers of bone turnover. RESULTS: Our results show that circulating OB precursors are more numerous and more immature in patients affected by fragility fractures than in osteoporotic patients without fractures. We also show that teriparatide treatment increases the expression of alkaline phosphatase and of OC in OB precursors; thus, it increases their degree of maturation. CONCLUSIONS: We suggest that teriparatide acts as anabolic agents also by promoting the maturation of OB precursors. PMID- 21617994 TI - Community mental health centres initiated by the South-Eastern Europe Stability Pact: evaluation in seven countries. AB - Eight community mental health care centres (initiated by the South-Eastern Europe Stability Pact) in Albania, Bosnia-Herzegovina, Croatia, Macedonia, Moldova, Montenegro and Romania were evaluated. Characteristics of patients, patient reported outcomes and patient views of care were assessed in 305 psychiatric patients. Patient characteristics varied across centres, with most patients having long term psychotic disorders. Treatment satisfaction and therapeutic relationships were rated favourably. Subjective quality of life mean scores were rather low, with higher satisfaction with health and dissatisfaction with the financial and employment situation. Being unemployed was the only factor associated with poor quality of life and lower treatment satisfaction. Most developing centres target patients with persistent psychotic disorders. Care appears highly valued by the patients. The findings encourage establishing more centres in the region and call for employment schemes for people with mental illnesses. PMID- 21617995 TI - A portable, integrated analyzer for microfluidic - based molecular analysis. AB - A portable, fully automated analyzer that provides actuation and flow control to a disposable, self-contained, microfluidic cassette ("chip") for point-of-care, molecular testing is described. The analyzer provides mechanical actuation to compress pouches that pump liquids in the cassette, to open and close diaphragm valves for flow control, and to induce vibrations that enhance stirring. The analyzer also provides thermal actuation for the temperature cycling needed for polymerase chain reaction (PCR) amplification of nucleic acids and for various drying processes. To improve the temperature uniformity of the PCR chamber, the system utilizes a double-sided heating/cooling scheme with a custom feedforward, variable, structural proportional-integral-derivative (FVSPID) controller. The analyzer includes a programmable central processing unit that directs the sequence and timing of the various operations and that is interfaced with a computer. The disposable cassette receives a sample, and it carries out cell lysis, nucleic acid isolation, concentration, and purification, thermal cycling, and either real time or lateral flow (LF) based detection. The system's operation was demonstrated by processing saliva samples spiked with B. cereus cells. The amplicons were detected with a lateral flow assay using upconverting phosphor reporter particles. This system is particularly suited for use in regions lacking centralized laboratory facilities and skilled personnel. PMID- 21617997 TI - Determination of total antioxidant capacity by a new spectrofluorometric method based on Ce(IV) reduction: Ce(III) fluorescence probe for CERAC assay. AB - A Ce(IV)-based reducing capacity (CERAC) assay was developed to measure the total antioxidant capacity (TAC) of foods, in which Ce(IV) would selectively oxidize antioxidant compounds but not citric acid and reducing sugars which are not classified as antioxidants. The method is based on the electron-transfer (ET) reaction between Ce(IV) ion and antioxidants in optimized acidic sulphate medium (i.e., 0.3 M H(2)SO(4) and 0.7 M Na(2)SO(4)) and subsequent determination of the produced Ce(III) ions by a fluorometric method. The fluorescent product, Ce(III), exhibited strong fluorescence at 360 nm with an excitation wavelength of 256 nm, the fluorescence intensity being correlated to antioxidant power of the original sample. The linear concentration range for most antioxidants was quite wide, e.g., 5.0 * 10(-7)-1.0 * 10(-5) M for quercetin. The developed procedure was successfully applied to the TAC assay of antioxidant compounds such as trolox, quercetin, gallic acid, ascorbic acid, catechin, naringin, naringenin, caffeic acid, ferulic acid, glutathione, and cysteine. The proposed method was reproducible, additive in terms of TAC values of constituents of complex mixtures, and the trolox equivalent antioxidant capacities (TEAC coefficients) of the tested antioxidant compounds gave good correlations with those found by reference methods such as ABTS and CUPRAC. PMID- 21617998 TI - A simple and sensitive fluorescence quenching method for the determination of H(2)O(2) using Rhodamine B and FE(3)O(4) nanocatalyst. AB - In pH 1.99 sodium acetate-HCl buffer solutions at 60 degrees C, Rhodamine B exhibited a strong fluorescence peak at 584 nm using an excitation wavelength of 548 nm. The fluorescence quenching occurred when Fe(3)O(4) nanoparticles catalyzed H(2)O(2) oxidation of Rhodamine B. Under the chosen conditions, the fluorescence intensity at 584 nm decreased when the concentration of H(2)O(2) increased. The fluorescence quenching intensity is linear with the concentration of H(2)O(2) in the range of 10-200 nmol/L. Thus, a new and simple and sensitive nanocatalytic fluorescence method was proposed for the determination of H(2)O(2) in synthetic sample, with satisfactory results. PMID- 21617996 TI - Elastomeric electrospun scaffolds of poly(L-lactide-co-trimethylene carbonate) for myocardial tissue engineering. AB - In myocardial tissue engineering the use of synthetically bioengineered flexible patches implanted in the infarcted area is considered one of the promising strategy for cardiac repair. In this work the potentialities of a biomimetic electrospun scaffold made of a commercial copolymer of (L)-lactic acid with trimethylene carbonate (P(L)LA-co-TMC) are investigated in comparison to electrospun poly(L)lactic acid. The P(L)LA-co-TMC scaffold used in this work is a glassy rigid material at room temperature while it is a rubbery soft material at 37 degrees C. Mechanical characterization results (tensile stress-strain and creep-recovery measurements) show that at 37 degrees C electrospun P(L)LA-co-TMC displays an elastic modulus of around 20 MPa and the ability to completely recover up to 10% of deformation. Cell culture experiments show that P(L)LA-co TMC scaffold promotes cardiomyocyte proliferation and efficiently preserve cell morphology, without hampering expression of sarcomeric alpha actinin marker, thus demonstrating its potentialities as synthetic biomaterial for myocardial tissue engineering. PMID- 21617999 TI - A novel rhodamine-benzimidazole conjugate as a highly selective turn-on fluorescent probe for Fe(3+). AB - In this manuscript, a novel probe RHBI based on the rhodamine-benzimidazole conjugate was designed and synthesized. RHBI showed an extreme selectivity for Fe(3+) over other metal ions such as Pb(2+), Ni(2+), Co(2+), Mn(2+), Zn(2+), Hg(2+), Cd(2+), Ag(+), Mg(2+), Ca(2+), Ba(2+), Na(+) and K(+) in acetonitrile. Upon the addition of 10 equiv. of Fe(3+), a 1098-fold fluorescence intensity enhancement was observed at the maximum emission wavelength of 582 nm. Both the Job's plot and ESI-MS showed that RHBI coordinated with Fe(3+) in a 1:1 stoichomitry and the calculated binding constant was 1.01 * 10(4) M(-1). The competition experiment for Fe(3+) ions mixed with other metal ions exhibited no obvious change except Cu(2+) that could induce a mild fluorescence quenching. Moreover, the fluorescence emission increased linearly with the Fe(3+) concentration in the range of 6 * 10(-6)-4 * 10(-5) M and the detection limit was 1.5 * 10(-8) M. PMID- 21618000 TI - Acute purulent pericarditis in childhood: don't forget beta-haemolytic group-A Streptococcus. PMID- 21618001 TI - Apocynin attenuates ventilator-induced lung injury in an isolated and perfused rat lung model. AB - RATIONALE: Apocynin suppresses the generation of reactive oxygen species (ROS) that are implicated in ventilator-induced lung injury (VILI). We thus hypothesized that apocynin attenuates VILI. METHODS: VILI was induced by mechanical ventilation with tidal volume (V(t)) of 15 ml/kg in isolated and perfused rat lung. Apocynin was administered in the perfusate at onset of mechanical ventilation. A group ventilated with low V(t) of 5 ml/kg served as control. Hemodynamics, lung injury indices, inflammatory responses, and activation of apoptotic pathways were determined upon completion of mechanical ventilation. RESULTS: There was an increase in lung permeability and lung weight gain after mechanical ventilation with high V(t), compared with low V (t). Levels of inflammatory cytokines including interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), and macrophage inflammatory protein-2 (MIP-2) increased in lung lavage fluids; concentrations of carbonyl, thiobarbituric acid reactive substances, and H(2)O(2) were higher in perfusates and lung lavage fluids, and expression of myeloperoxidase, JNK, p38, and caspase-3 in lung tissue was greater in the high-V(t) than in the low-V(t) group. Administration of apocynin attenuated these inflammatory responses and lung permeability associated with decreased activation of nuclear factor-kappaB. CONCLUSIONS: VILI is associated with inflammatory responses including generation of ROS, cytokines, and activation of mitogen-activated protein kinase cascades. Administration of apocynin at onset of mechanical ventilation attenuates inflammatory responses and VILI in the isolated, perfused rat lung model. PMID- 21618002 TI - Effect of statin therapy on plasma adiponectin concentrations in patients with the sepsis syndrome: a preliminary investigation. PMID- 21618003 TI - What are the different initial presentations of frontotemporal dementia? AB - Frontotemporal dementia (FTD) symptoms at the beginning of illness are in either the realm of behavioral disturbance or in language disruption, also known as aphasia. Based on specific constellations of behavioral change or characteristics of the aphasia, physicians can anticipate the type of protein that is abnormal in the brain. Family history rich with similar instances of illness can further guide the cause of illness in an individual. Knowing the type of protein abnormality (proteinopathy) from among the three most common in FTD (tau, TDP-43, or fused in sarcoma) can help treating clinicians to advise families on the future course of illness and future clinical drug trials that would be most applicable to each patient. PMID- 21618004 TI - New potential therapeutic approaches in frontotemporal dementia: oxytocin, vasopressin, and social cognition. AB - Frontotemporal dementias are neurodegenerative disorders marked by mid-life onset and progressive changes in behavior, decision making, and/or language. In several subtypes of frontotemporal dementia (FTD), damage to regions of the frontal and temporal lobes that occurs early in the disease course critically impairs emotional processing, social cognition, and behavior. To date, there are no curative or disease-modifying treatments for FTD, nor any approved treatments for the loss of empathy, impaired social cognition, and behavior of patients with FTD. A growing body of research suggests that the neuropeptides oxytocin and vasopressin are important mediators of social behavior in humans and other animals. Here, we review the effects of oxytocin and vasopressin on social cognition and behavior and discuss the potential implications for these agents as novel treatments in FTD. PMID- 21618005 TI - A discriminant analysis of trace elements in scalp hair of healthy controls and stage-IIIB non-small cell lung cancer (NSCLC) patients. AB - Our work aimed at extending the search for the trace elements (TE) abnormalities in patients with lung cancer and in healthy controls who smoke, and also for evidence of a possible association between lung cancer and TE. The analysis of the hair from patients with Stage-IIIB non-small cell lung cancer (group 1) and healthy controls (group 2) were analyzed using the inductively coupled plasma mass spectrometry technique in order to obtain information on the correlation between the lung cancer patients and healthy controls. Sixty-seven one-hair samples in group 1 were individually collected before chemoradiotherapy. For comparison, 74 hair samples were collected from group 2. In group 1, the trace elements present at the highest levels were measured to be Ca, Zn, Sn, Na and Mg, respectively, and they were quantified as 68.2, 53.2, 33.9, 23.3, and 28.9 MUg.kg(-1), respectively. In group 2, the trace elements present at the highest levels were Zn, Mg, Ca, Fe, and Se, respectively, and they were quantified as 109.7, 31.9, 30.8, 25.0, and 20.1 MUg.kg(-1). In group 1, the highest levels of Ca, Sn, and Na were 2.03, 1.06, and 1.01 times higher, respectively, compared with group 2. In group 2, Zn, Mg, Fe, and Se were 2, 1.01, 2.7, and 1.6 times higher, respectively, compared with group 1. When the levels of trace elements were compared between groups 1 and 2 using Student's t test, the levels of Ag, Au, Be, Bi, Ca, Cd, Ce, Co, Cr, Cu, Fe, Ga, Hg, K, Ni, Rb, Rh, Sb, Sc, Ti, V, and Zn were found to be statistically different (p < 0.05). According to Pearson's correlation, the most powerful correlation was found for Cr-As (r = 0.858) couple in group 1 (r = 0.745) and for Mn-Cr couple in group 2. The factors obtained according to converted matrix were observed to be as follows: for group 1, first factor, ten variables (Cd, Li, Cs, Ag, Rb, Pd, Ga, Zn, Al, and K); second factor, seven variables (Cr, As, Sn, Co, Ca, Rh, and Fe) and third factor, four variables (Mn, Au, Cu, and Hg). Within the first factor that best describes the overall change, the most important variables are Cd and Li, respectively the first and the second factors. Group 2 contained the following: first factors, six variables, Cr, Mn, Al, Ba, Rb, and Pb; second factor, eight variables, Co, As, Sn, Cd, Hg, Cs, Ca, and Ce; third factor, five variables Na, Ga, Be, B, and Sr; and fourth factor, two variables K and Ag. First, second, third, and fourth factors explain the 36% of the overall change. Within the first factor that best described the overall change, the most important variables were Cr and Mn. In this analysis, we observed that the group 2 trace elements accumulated were heavy metals and that the control group showed both heavy metals and macroelements required for the body. The average trace elements levels in the two groups were evaluated. In addition, the general role of trace elements in the lung carcinogenic processes was discussed. The study revealed that the carcinogenic processes are significantly affecting the trace elements and the trace elements distribution in the hair of lung cancer patients compared with the healthy controls. It was revealed that there was a relation between lung cancer and trace elements, especially heavy metals. Our findings suggest that the heavy metals accumulated in the body may pose a high risk for lung cancer development. PMID- 21618006 TI - Reduction of Sb(V) in a human macrophage cell line measured by HPLC-ICP-MS. AB - Drugs based on pentavalent antimony are first-line treatment of the parasite disease leishmaniasis. It is generally believed that Sb(V) acts as a prodrug, which is activated by reduction to Sb(III); however, the site of reduction is not known. It has been hypothesised that the reduction takes place in the parasites' host cells, the macrophages. In this study, the human macrophage cell line Mono Mac 6 was exposed to Sb(V) in form of the drug sodium stibogluconate (PentostamTM). Cell extracts were analysed for Sb species by high-performance liquid chromatography with inductively coupled plasma-mass spectrometry detection. We found that Sb(V) is actually reduced to Sb(III) in the macrophages; up to 23% of the intracellular Sb was found as Sb(III). Transfer of the cells to Sb-free medium rapidly decreased their Sb(V) and Sb(III) content. Induction of the cell's production of reactive oxygen species did not have any marked effect on the intracellular amounts of Sb(III). PMID- 21618007 TI - [X, Y,... lack of personnel?]. PMID- 21618008 TI - Visualisation of the chemical space of fragments, lead-like and drug-like molecules in PubChem. AB - The 4.5 million organic molecules with up to 20 non-hydrogen atoms in PubChem were analyzed using the MQN-system, which consists in 42 integer value descriptors of molecular structure. The 42-dimensional MQN-space was visualised by principal component analysis and representation of the (PC1, PC2), (PC1, PC3) and (PC2, PC3) planes. The molecules were organized according to ring count (PC1, 38% of variance), the molecular size (PC2, 25% of variance), and the H-bond acceptor count (PC3, 12% of variance). Compounds following Lipinski's bioavailability, Oprea's lead-likeness and Congreve's fragment-likeness criteria formed separated groups in MQN-space visible in the (PC2, PC3) plane. MQN similarity searches of the 4.5 million molecules (see the browser available at www.gdb.unibe.ch ) gave significant enrichment factors for recovering groups of fragment-sized bioactive compounds related to ten different biological targets taken from Chembl, allowing lead-hopping relationships not seen with substructure fingerprint similarity searches. The diversity of different compound series was analyzed by MQN-distance histograms. PMID- 21618009 TI - Visualisation and subsets of the chemical universe database GDB-13 for virtual screening. AB - The chemical universe database GDB-13, which enumerates 977 million organic molecules up to 13 atoms of C, N, O, S and Cl following simple chemical stability and synthetic feasibility rules, represents a vast reservoir for new fragments. GDB-13 was classified using the MQN-system discussed in the preceding paper for the analysis of PubChem fragments. Two hundred and fifty-five subsets of GDB-13 were generated by the combinatorial use of eight restrictive criteria, including fragment-like ("rule of three") and scaffold-like (no acyclic carbon atoms) filters. Virtual screening for analogs of 15 commercial drugs of 13 non-hydrogen atoms or less shows that retrieving MQN-neighbors of a query molecule from GDB-13 or its subsets provides on average a 38-fold enrichment in structural analogs (Daylight-type substructure fingerprint Tanimoto T (SF) > 0.7), and a 75-fold enrichment in shape-similar analogs (ROCS TanimotoCombo score > 1.4). An MQN searchable version of GDB-13 is provided at www.gdb.unibe.ch . PMID- 21618010 TI - Legacy effects overwhelm the short-term effects of exotic plant invasion and restoration on soil microbial community structure, enzyme activities, and nitrogen cycling. AB - Plant invasions can have substantial consequences for the soil ecosystem, altering microbial community structure and nutrient cycling. However, relatively little is known about what drives these changes, making it difficult to predict the effects of future invasions. In addition, because most studies compare soils from uninvaded areas to long-established dense invasions, little is known about the temporal dependence of invasion impacts. We experimentally manipulated forest understory vegetation in replicated sites dominated either by exotic Japanese barberry (Berberis thunbergii), native Viburnums, or native Vacciniums, so that each vegetation type was present in each site-type. We compared the short-term effect of vegetation changes to the lingering legacy effects of the previous vegetation type by measuring soil microbial community structure (phospholipid fatty acids) and function (extracellular enzymes and nitrogen mineralization). We also replaced the aboveground litter in half of each plot with an inert substitute to determine if changes in the soil microbial community were driven by aboveground or belowground plant inputs. We found that after 2 years, the microbial community structure and function was largely determined by the legacy effect of the previous vegetation type, and was not affected by the current vegetation. Aboveground litter removal had only weak effects, suggesting that changes in the soil microbial community and nutrient cycling were driven largely by belowground processes. These results suggest that changes in the soil following either invasion or restoration do not occur quickly, but rather exhibit long-lasting legacy effects from previous belowground plant inputs. PMID- 21618011 TI - A gall-inducing arthropod drives declines in canopy tree photosynthesis. AB - Mature forest canopies sustain an enormous diversity of herbivorous arthropods; however, with the exception of species that exhibit large-scale outbreaks, canopy arthropods are thought to have relatively little influence on overall forest productivity. Diminutive gall-inducing mites (Acari; Eriophyoidae) are ubiquitous in forest canopies and are almost always highly host specific, but despite their pervasive occurrence, the impacts of these obligate parasites on canopy physiology have not been examined. We have documented large declines in photosynthetic capacity (approx. 60%) and stomatal conductance (approx. 50%) in canopy leaves of mature sugar maple (Acer saccharum) trees frequently galled by the maple spindle gall mite Vasates aceriscrumena. Remarkably, such large impacts occurred at very low levels of galling, with the presence of only a few galls (occupying approx. 1% of leaf area) compromising gas-exchange across the entire leaf. In contrast to these extreme impacts on the leaves of adult trees, galls had no detectible effect on the gas-exchange of maple saplings, implying large ontogenetic differences in host tolerance to mite galling. We also found a significant negative correlation between canopy tree radial increment growth and levels of mite galling. Increased galling levels and higher physiological susceptibility in older canopy trees thus suggest that gall-inducing mites may be major drivers of "age-dependent" reductions in the physiological performance and growth of older trees. PMID- 21618012 TI - Study of the left renal variations by means of magnetic resonance imaging. AB - PURPOSE: A prospective study was designed to figure out the percentages of the left renal vein variations using routine lumbar spinal magnetic resonance imaging (MRI) in patients with neurological problems. METHODS: Between March 2010 and October 2010, the study population was recruited from a total of 2,644 consecutive patients who would undergo a routine lumbar spinal MRI examination. In addition to the routine MRI sequences, axial, balanced turbo field echo sequence (BTFE-BH SENSE) was applied after detection of a left renal vein variation. RESULTS: As the left renal vein variations, retroaortic left renal vein (RLRV) and circumaortic left renal vein were detected. The number of cases with the correspondent percentages of the total left renal vein variations, RLRV and circumaortic left renal vein were 71/2,644 (2.68%), 44/2,644 (1.66%) and 27/2,644 (1.02%), respectively. In cases with RLRV, the numbers of males and females with their correspondent percentages were 19/44 (43.2%) and 25/44 (56.8%), respectively. In cases with circumaortic left renal vein, the numbers of males and females with their correspondent percentages were 13/27 (48.1%) and 14/27 (51.9%), respectively. With Chi-square test, no statistically significant gender difference was found between the percentages of left renal vein variations (P = 0.83). CONCLUSIONS: MRI is useful in detecting RLRV and circumaortic left renal vein. If a left renal vein variation is detected, an additional BTFE-BH SENSE sequence is suggested to confirm whether it is retroaortic or circumaortic. PMID- 21618013 TI - The retinacula of Weitbrecht in the adult hip. AB - PURPOSE: The aim of the study was to describe the retinacula of Weitbrecht in the adult hip. MATERIALS AND METHODS: Specimens were obtained from 30 adult hips, average age was 77 years (age range 43-91 years), 8 specimens were fixed by formalin solution and 22 were not fixed. RESULTS: Anterior retinaculum was found in 40% of examined specimens. The anterior retinaculum was in 83% of cases formed by a flat plate and in 17% by two to three parallel bands. Medial retinaculum was present constantly, extending from the attachment of the articular capsule at the base of the lesser trochanter towards the fovea capitis femoris as far as the edge of the articular cartilage. Typically, the retinaculum had the form of an inverted "T". Of the three retinacula, the medial one was the strongest. Lateral retinaculum was also present constantly. In 89% of cases, it had the form of a quadrilateral plate adjacent to the upper surface of the femoral neck. This plate arises from the insertion of the articular capsule on the upper part of the femoral neck at the base of the greater trochanter close to the trochanteric fossa. The plate extended along the upper edge of the femoral neck as far as the edge of the articular cartilage. Microscopic examination revealed fine blood vessels running through the retinacula. CONCLUSION: Lateral retinaculum and medial retinaculum are constant synovial plicae in terms of both occurrence and localization. Nutritive arteries run through both the plicae to supply the femoral head. PMID- 21618014 TI - 3D Modeling of branching vessels from anatomical sketches: towards a new interactive teaching of anatomy: Interactive virtual blackboard. AB - Sketching is an intuitive way to explain spatial relationships between complex objects. The French community of Anatomists are used to teaching didactic lectures on a blackboard with their colored chalks. The increasing complexity of the sketches affords to the students an opportunity to work out a mental representation of anatomical structures in 3D. To help students perform this labored step, we present a new interactive blackboard which constructs plausible 3D models of branching vessels from a single sketch. We exploit the sketching conventions used in anatomical drawings to infer depth and curvature. We then model the set of branching vessels as a convolution surface generated by a graph of skeleton curves. Classic situations, focused on arteries, have been analyzed to manage vessels' curvatures, subdivisions and overlaps. Original sketches and 3D models are presented for each case. No specific training is required to use the interface. The anatomists have begun to embrace a new generation of 3D digital modeling applications as tools for anatomical teaching. We discuss the future use of this system as a step towards the interactive teaching of anatomy. PMID- 21618015 TI - Reliability of magnetic resonance imaging measurements of the cross-sectional area of the muscle contractile and non-contractile components. AB - BACKGROUND/PURPOSE: Magnetic resonance imaging (MRI) is increasingly used to study skeletal muscles patients with muscular disorders. We report an MRI technique for evaluating the trunk muscles. This technique takes both the component surface area (CSA) and the density of the muscles on MRI axial slices into account . Using a computer-based image analysis system, we combined MRI data measuring the muscle CSA, which was separated into the contractile component (CCSA) and the non-contractile component (NCCSA). The purpose of this study was to analyze the reliability of this method of measuring the CSA, CCSA and NCCSA in trunk muscles on MRI axial slices through L4 and T12. METHODS: Thirty volunteer subjects were enrolled in this study. Two acquisitions were performed. For the reliability analyses, each of the two slices (T12 and L4) from 30 subjects was measured by three raters trained in this technique, on two occasions 2 weeks apart. Each muscle was surrounded and its CSA, NCCSA and CCSA were recorded. For each muscle, the agreement between the two sets of 30 measurements performed by three observers was evaluated by calculating an intra-class correlation coefficient (ICC). RESULTS: Regarding the slice through L4 and T12, the reliabilities of the measurement of CSA, CCSA were very good for all the muscles except the parietal muscles. CONCLUSION: MRI measurements of the trunk muscle cross-sectional areas and of the CCSA and NCCSA are reliable. PMID- 21618016 TI - Seromucinous hamartoma of the nasal cavity: a report of two cases and review of the literature. AB - The sinonasal tract is a complex anatomic site, home to a wide variety of reactive, inflammatory, benign, and malignant lesions. Inflammatory polyps and papillomas are usually easily recognized by pathologists. A poorly understood lesion that has been more clearly defined in recent years is the nasal hamartoma. The epithelial subtypes include seromucinous hamartoma, respiratory epithelial adenomatoid hamartoma, and hybrid lesions. Seromucinous hamartomas have only been recognized and substantially reported over the past few years. They are a diagnostic challenge, needing to be distinguished from low grade adenocarcinomas, and are of interest because most of the basic questions about their pathophysiology remain unanswered. Herein, we present two novel cases of seromucinous hamartoma with features that partly expand the morphologic spectrum of these lesions, discuss the differential diagnosis, and review the literature to compare our findings with previously reported cases with the aim of better understanding this interesting entity. PMID- 21618017 TI - Acetyl-L-carnitine prevents carbon tetrachloride-induced oxidative stress in various tissues of Wistar rats. AB - Acetyl-L-carnitine (ALCAR) has been shown to prevent experimental selenite cataractogenesis, a manifestation of oxidative stress, but little is known about its potential in other settings of oxidative stress. The present study was based on the hypothesis that ALCAR prevents carbon tetrachloride (CCl(4))-induced oxidative stress in vital tissues. Male albino Wistar rats were divided into three groups, each of six rats. Group I (control) rats received only vehicle (1 ml/kg b.w.) for 4 days; Group II (CCl(4)-exposed, untreated) rats received CCl(4) (2 ml/kg b.w.) on the second and third days and vehicle on the first and fourth days; Group III (CCl(4)-exposed, ALCAR-treated) rats received ALCAR (200 mg/kg b.w.) for 4 days and CCl(4) on the second and third days. All administrations were made intraperitoneally. After the experimental period, significantly (P < 0.05) elevated mean serum levels of aspartate transaminase, alanine transaminase, alkaline phosphatase, and lactate dehydrogenase were observed in Group II rats when compared to Group I and Group III rats. The mean levels of vitamin C, vitamin E, and reduced glutathione and the mean activities of superoxide dismutase, catalase, and glutathione peroxidase were significantly (P < 0.05) lower in samples of hemolysate and of liver, kidney, and brain tissues of Group II rats than those in Group I and Group III rats. The mean level of lipid peroxidation was significantly (P < 0.05) higher in Group II rats than that in Group I and Group III rats. Moreover, the CCl(4)-induced upregulation of inducible nitric oxide synthase expression was prevented by ALCAR in the liver and brain tissues. These results suggest that ALCAR is able to prevent the CCl(4) induced oxidative stress. PMID- 21618018 TI - relaxGUI: a new software for fast and simple NMR relaxation data analysis and calculation of ps-ns and MUs motion of proteins. AB - Investigation of protein dynamics on the ps-ns and MUs-ms timeframes provides detailed insight into the mechanisms of enzymes and the binding properties of proteins. Nuclear magnetic resonance (NMR) is an excellent tool for studying protein dynamics at atomic resolution. Analysis of relaxation data using model free analysis can be a tedious and time consuming process, which requires good knowledge of scripting procedures. The software relaxGUI was developed for fast and simple model-free analysis and is fully integrated into the software package relax. It is written in Python and uses wxPython to build the graphical user interface (GUI) for maximum performance and multi-platform use. This software allows the analysis of NMR relaxation data with ease and the generation of publication quality graphs as well as color coded images of molecular structures. The interface is designed for simple data analysis and management. The software was tested and validated against the command line version of relax. PMID- 21618019 TI - [Glaucoma surgery in childhood]. AB - Technical characteristics and a long-term therapeutic strategy due to a long life expectancy play a key role in pediatric glaucoma surgery. The well-established angle surgery (goniotomy and trabeculotomy) achieves successful results in primary childhood glaucoma. Trabeculectomy seems to have been displaced as a secondary approach by glaucoma drainage devices (GDD) in primary childhood glaucoma due to inferior results, especially for children under 3 years of age. Even for secondary childhood glaucoma the results of GDD are encouraging, especially for therapy refractory aphakic glaucoma. In the first 2 years after GDD surgery success rates are about 80% for pediatric glaucoma and the results appear to be independent of the type of glaucoma and implant used. The complications of GDD are balanced to the faster intraocular pressure (IOP) control during the phase of visual acuity development. Cyclodestructive procedures may be applied as a secondary adjuvant approach but they increase the risk of conjunctival scarring and hypotony for subsequent procedures. PMID- 21618020 TI - XRD and DFT-modelized structures of a pteridine-2,4(1H,3H)-dithione/N,N' dimethylformamide H-bonded cluster (2:2). MO study of the coordination ability. AB - The title compound, C(6)H(4)N(4)S(2).C(3)H(7)NO, crystallizes in the monoclinic space group C 2/c with a = 26.673(5), b = 5.397(1), c = 16.522(3) A, beta = 95.49(3) degrees , Z = 8, R = 0.0461 for 1891 reflections with I > 2sigma(I) and 174 parameters (4 restraints). Single pteridine-2,4(1 H,3 H)-dithione and dimethylformamide molecules are packed via N-H...O and N-H...N hydrogen bonds into centrosymmetric clusters containing two molecules of each class; these are roughly planar and placed into two different sets of planes -both containing the [-1,0,2] direction- mutually angled by 77.8 degrees . Despite the distance between two neighbor planes in each set is ca. 3.4 A, the analysis of pi,pi stacking interactions shows too large slippage distance between aromatic rings from contiguous planes. Additional sigma-pi interactions between S2, S4 and O1S atoms and pyrazine or pyrimidine rings from adjacent molecules are present. The structure for the cluster [DTLM-DMF](2) has been simulated by using the density functionals B1B95 (6-31 G(d) and 6-31+G(d) basis sets) and M06-2X (6-31 G(d) basis set). As a result, the M06-2X/6-31 G(d) approach provides the best agreement with the experimental XRD data. For a better evaluation of the intermolecular interactions, the superposition of two dimeric adducts [DTLM DMF](2) has been modelized. The binding capability of DTLM ligand was simulated on systems containing two metal-binding modes to palladium (N5-S4 and N1-S2) with different chelate size. The analysis of the frontier orbitals points out that the link with the metallic centers will take place through the sulfur atoms. PMID- 21618021 TI - Four-component pharmacophore model for endomorphins toward MU opioid receptor subtypes. AB - In the present work, a series of simulation tools were used to determine structure-activity relationships for the endomorphins (EMs) and derive MU pharmacophore models for these peptides. Potential lowest energy conformations were determined in vacuo by systematically varying the torsional angles of the Tyr(1)-Pro(2) (omega(1)) and Pro(2)-Trp(3)/Phe(3) (omega(2)) as tuning parameters in AM1 calculations. These initial models were then exposed to aqueous conditions via molecular dynamics simulations. In aqueous solution, the simulations suggest that endomorphin conformers strongly favor the trans/trans pair of the omega(1)/omega(2) amide bonds. From two-dimensional probability distributions of the ring-to-ring distances with respect to the pharmacophoric angles for EMs, a selectivity range of MU(1) is ca. 8.3 ~ 10.5 A for endomorphin-2 and selectivity range of MU(2) is ca. 10.5 ~ 13.0 A for endomorphin-1 were determined. Four component MU-pharmacophore models are proposed for EMs and are compared to the previously published delta- and kappa-pharmacophore models. PMID- 21618022 TI - A viral grandfather: genomics in 2010 contradict Darwin's vision of evolution. PMID- 21618023 TI - Mechanism of rate enhancement of wood fiber saccharification by cationic polyelectrolytes. AB - Cationic polyelectrolytes can increase the cellulase-induced hydrolysis rates of bleached wood fiber. We show that the polymer associates mainly with the amorphous region of fiber and acts principally on endoglucanase. Fiber/water partitioning of the enzyme follows a Langmuir isotherm for the untreated fiber but a Freundlich isotherm is obeyed for the polymer-treated fiber. PMID- 21618024 TI - Immobilized enzymes affect biofilm formation. AB - The effect of the activity of immobilized enzymes on the initial attachment of pathogenic bacteria commonly associated with nosocomial infections (Pseudomonas aeruginosa and Staphylococcus epidermidis) was investigated. The proteolytic enzymes, subtilisin A and the glycoside hydrolase cellulose, were covalently attached onto poly(ethylene-alt-maleic) anhydride copolymer films. A comparison between active and heat-inactivated surfaces showed that while the activity of immobilized cellulase reduced the attachment of S. epidermidis by 67%, it had no effect on the attachment of P. aeruginosa. Immobilized subtilisin A had opposite effects: the active enzyme had no effect on the attachment of S. epidermidis but reduced the attachment of P. aeruginosa by 44%. The results suggest that different biomolecules are involved in the initial steps of attachment of different bacteria, and that the development of broad-spectrum antifouling enzymatic coatings will need to involve the co-immobilization of enzymes. PMID- 21618025 TI - Expressions of thermostable bacterial cellulases in tobacco plant. AB - An economical method for the conversion of lignocellulosic biomass is to use plants as bioreactors for cellulases production. Two bacterial thermostable cellulases (E2 and E3) and a E3-E2 fusion form were expressed in tobacco, driven by a double 35S promoter and 5' TEV-UTL. The enzymes were targeted to the apoplast and cytosol via 5' signal peptides and 3' retention signal peptides, respectively, and all showed functional activities. All transgenic plants exhibited normal growth compared to wild type. Transgenic plants that expressed apoplast-localized E2 had the highest average activity, about 1.5 and 3 times higher than those expressed ER-localized and cytosolic E2, respectively. Effect of subcellular compartment localization was due primarily to post-transcriptional modification, since mRNA abundances were similar despite the range of cellulase activities obtained. The recombinant cellulases exhibited good thermostability below 65 degrees C. After storing for 3 days at -20 and 28 degrees C, the enzymes lost nearly 20 and 80% of activity, respectively. The results suggested a potential application for heterologous expression of cellulases in plant for biomass conversion. PMID- 21618026 TI - Transcatheter closure of perimembranous ventricular septal defects (VSD) with VSD occluder: early and mid-term results. AB - Results of perimembranous ventricular septal defects (pmVSD) transcatheter closure have been reported in the literature, mostly with the Amplatzer VSD device (muscular or eccentric) (AGA Medical Corp., Golden Valley, MN, USA). However, the data of percutaneous closure of pmVSD with VSD occluder (VSD-O) made in China are still limited. We sought to analyze the safety, efficacy, and follow up results of percutaneous closure of pmVSD with VSD-O made in China. Seventy eight patients underwent percutaneous closure of pmVSD at our institution between February 2005 and June 2007. A VSD device made in china (Huayishengjie Medical Corp., Beijing, China) was used in all subjects. The mean age at closure was 11 years (range 2.5-44 years). The attempt to place the device was successful in 74 patients (94.9%). The median device size used was 8 mm (range 5-16 mm). No deaths occurred. Total occlusion rate was 62.8% at completion of the procedure, rising to 87.2% at discharge and 99% during the follow-up. A total of eight early complications occurred (10.3%), but in all subjects these were transient. The median follow-up was 32 months. The most significant complication was complete atrioventricular block (cAVB) in the early phase (five subjects, 6.4%) and during the follow-up (one subject, 1.3%), and there was no need for pacemaker implantation in six subjects. Logistic regression analysis showed that the only variable significantly associated with the occurrence of this complication was age at the time of the procedure (p = 0.025; OR 0.22). All subjects experiencing this problem were <5 years old. Percutaneous pmVSD closure used VSD-O made in China is associated with excellent success and closure rates, no mortality, and low morbidity. Nowadays, pmVSD percutaneous closure is a valuable alternative to surgery. Longer follow-up data and improvements in device characteristics are needed to reduce the risk of cAVB. PMID- 21618027 TI - A miR-21 inhibitor enhances apoptosis and reduces G(2)-M accumulation induced by ionizing radiation in human glioblastoma U251 cells. AB - MicroRNAs (miRNAs) are small noncoding RNAs that take part in diverse biological processes by suppressing target gene expression. Elevated expression of miR-21 has been reported in many types of human cancers. Radiotherapy is a standard adjuvant treatment for patients with glioblastoma. However, the resistance of glioblastoma cells to radiation limits the success of this treatment. In this study, we found that miR-21 expression was upregulated in response to ionizing radiation (IR) in U251 cells, which suggested that miR-21 could be involved in the response of U251 cells to radiation. We showed that a miR-21 inhibitor enhanced IR-induced glioblastoma cell growth arrest and increased the level of apoptosis, which was probably caused by abrogation of the G(2)-M arrest induced by IR. Further research demonstrated that the miR-21 inhibitor induced the upregulation of Cdc25A. Taken together, these findings suggest that miR-21 inhibitor can increase IR-induced growth arrest and apoptosis in U251 glioblastoma cells, at least in part by abrogating G(2)-M arrest, and that Cdc25A is a potential target of miR-21. PMID- 21618028 TI - Background enhancement of mammary glandular tissue on breast dynamic MRI: imaging features and effect on assessment of breast cancer extent. AB - BACKGROUND: Just as mammographic breast density influences mammographic sensitivity, the degree of background enhancement in breast magnetic resonance imaging (MRI) may influence the sensitivity of breast MRI. The purpose of this study is to assess the influence of background enhancement on the accuracy of breast cancer extent assessment using MRI and to assess the correlation between the accuracy of breast cancer extent assessment and the kinetic analysis of background enhancement in dynamic contrast-enhanced MRI. METHODS: Seventy bilateral breast MRI examinations were evaluated to assess the extent of a known primary tumor. Background enhancement was classified into four categories by visual assessment: minimal, mild, moderate, and marked, in the early dynamic phase and in the late dynamic phase. The correlation of the results with histological findings was examined. RESULTS: Background enhancement grade showed a significant tendency to increase during dynamic contrast-enhanced MRI. When classifying background enhancement at early dynamic phase, the accuracy of tumor extent assessment by MRI with moderate/marked background enhancement was 60%, which was lower than the 78% accuracy with minimal/mild background enhancement, but not significantly so (p = 0.153). When classifying background enhancement at late dynamic phase, the accuracy with moderate/marked background enhancement was 61%, which was significantly lower than the 83% accuracy with minimal/mild background enhancement (p = 0.034). There was no tumor-size-related bias between the groups (p = 0.089). CONCLUSION: The degree of background enhancement on breast MRI affects the accuracy of breast cancer extent assessment, especially at late dynamic phase. PMID- 21618029 TI - Isolation and characterization of the first Chinese porcine sapelovirus strain. AB - Investigations were carried out to identify the causative agent of acute diarrhea, respiratory distress, and polioencephalomyelitis of pigs on a swine farm in Shanghai, China. Samples from the affected animals were tested for viruses and bacteria that are known to cause similar symptoms in swine, and only porcine sapelovirus (PSV; designated as csh strain) was isolated. The presence of PSV was further confirmed by the specific cytopathic effects observed in susceptible cells and by the results of PCR and electron microscopy. Nucleotide sequencing and phylogenetic analysis showed that this isolate is PSV. When inoculated into healthy pigs, PSV.csh caused the same symptoms as observed in the affected herd. Therefore, PSV.csh is the causative agent of this disease. To the best of our knowledge, this is the first report of PSV infecting piglets in China. PMID- 21618030 TI - Complete sequence and genetic characterization of pigeon avian nephritis virus, a member of the family Astroviridae. AB - In the current study, the complete genome sequence of a member of the family Astroviridae isolated from pigeons was determined through genetic characterization and phylogeny analysis. The isolated genome sequence was proposed to be that of pigeon avian nephritis virus (ANV), whose genome structure and characteristics were similar to previously reported avian astroviruses. The sequenced ssRNA genome comprises 6928 nucleotides, excluding the poly(A) tail, and contains three open reading frames. Phylogenetic analysis using a partial nucleotide sequence of the polymerase gene and the entire amino acid sequence of the full-length capsid protein revealed that pigeon avian nephritis virus is closely related to the previously published ANV, especially to the Japanese G 4260 and Chinese strains. This investigation provides information on the sequence and genetic characteristics of this virus and contributes to a better understanding of pigeon ANV and the possible occurrence of astrovirus transmission between chickens and pigeons. PMID- 21618031 TI - Immune Cells in Colorectal Cancer: Prognostic Relevance and Role of MSI. AB - There is growing evidence that both local and systemic inflammatory responses play an important role in the progression of a variety of solid tumors. Colorectal cancer (CRC) results from the cumulative effect of sequential genetic alterations, leading to the expression of tumor-associated antigens possibly inducing a cellular anti-tumor immune response. It is well recognized that cytotoxic lymphocytes (CTLs) constitute one of the most important effector mechanisms of anti-tumor-immunity. However, their potential prognostic influence in CRC remains controversial. In addition, other key players like natural killer cells, tumor associated macrophages and regulatory T cells play an important role in the immune attack against CRC and need further investigation. This review will mainly focus on the role of the adaptive immune system in CRC and particularly in regard to microsatellite instability. PMID- 21618032 TI - Epicrania fugax with backward radiation: clinical characteristics of nine new cases. AB - Epicrania fugax (EF) is a novel syndrome, described as a paroxysmal and brief head pain, starting in posterior cranial regions and rapidly spreading forward ipsilateral eye, nose or forehead. Two patients with comparable clinical features stemming from frontal scalp to ipsilateral posterior regions have been recently described and proposed as backward radiation epicrania fugax (BREF). We report a new series of nine BREF and compare their clinical characteristics with 18 forward radiation EF (FREF). Since first description of BREF in February 2010 we have assessed nine patients (four males, five females) with this clinical picture at an outpatient headache office in a Tertiary Hospital. Comparison is established with 18 FREF patients (6 males, 12 females), attended since the publication of first series of EF in March 2008. We found no differences between BREF and FREF, respectively, in age at onset (43.4 +/- 13.1 vs. 42.5 +/- 17.7 years), female/male ratio (5/4 vs. 12/6), pain intensity (6.9 +/- 2.1 vs. 6.8 +/- 2.1 in a 0-10 visual analogical scale), duration (7.1 +/- 4.9 vs. 5.7 +/- 4.3 s) and frequency of episodes per day (7 +/- 8.4 vs. 9.9 +/- 15.4). Patients in BREF group presented less frequently interictal pain in stemming point (22.2 vs. 55.5%) and accompanying autonomic signs (33.3 vs. 55.5%), but without statistical significance in both the cases. This series reinforces the proposal of EF as a new headache variant or a new headache syndrome. Clinical picture of brief pain paroxysms starting in the anterior scalp and radiating backwards does not fit known headaches or neuralgias and might correspond to a reverse variant of EF, clinical characteristics of which are comparable to FREF. PMID- 21618033 TI - 3, 6 or 9-band shooters: tailor made ligators as needed. PMID- 21618034 TI - Intergenerational effect of juvenile hormone on offspring in Pogonomyrmex harvester ants. AB - Parents can influence the phenotypes of their offspring via a number of mechanisms. In harvester ants, whether female progeny develop into workers or daughter queens is strongly influenced by the age and temperature conditions experienced by their mother, which is associated with variation in maternal ecdysteroid deposition in fertilized eggs. In many insects, juvenile hormone (JH) is antagonistic to ecdysteroid release, suggesting that seasonal and age-based variation in maternal JH titers may explain maternal effects on offspring size and reproductive caste. To test this hypothesis, we artificially increased maternal JH titers with methoprene, a JH analog, in laboratory colonies of two Pogonomyrmex populations exhibiting genetic caste determination. Increasing maternal JH resulted in a 50% increase in worker body size, as well as a sharp reduction in total number of progeny reared, but did not alter the genotype of progeny reared to adulthood. The intergenerational effect of JH manipulation was not mediated by a reduction in ecdysteroid deposition into eggs; instead, changes in egg size, trophic egg availability or brood/worker ratio may have altered the nutritional environment of developing larvae. Egg ecdysteroid content was significantly negatively correlated with natural variation in worker body size, however, suggesting that there are multiple independent routes by which queens can modify offspring phenotypes. PMID- 21618035 TI - White light-mediated Cu (II)-5FU interaction augments the chemotherapeutic potential of 5-FU: an in vitro study. AB - 5-Fluorouracil (5-FU) is a potent photosensitizer used in colon and rectal cancers. 5-FU on galvanostatic electrolysis or radiation-induced oxidation of aqueous solution yields N(1)-C(5)-linked dimmer hydrate of 5-FU. Copper is presently associated with chromatin; in cancer cells the concentration of copper is very high. It has been shown to be capable of mediating the action of several anticancer drugs through the production of reactive oxygen species (ROS). The objective of the present study is to determine the Cu (II)-mediated anticancer mechanism of 5-FU under photo-illumination as well as 5-FU alone. We have shown that a pro-oxidant action was enhanced when Cu (II) was used with 5-FU as compared to 5-FU alone. This may be due to the inhibition of dimerization of 5-FU when present in combination with Cu (II) under photo-illumination. It was also shown that 5-FU alone as well as in combination with Cu (II) was able to generate oxidative stress in lymphocyte which is inhibited by scavengers of ROS. Moreover, the results of Fourier-transformed infrared spectra lead to the conclusion that the dimerization of 5-FU was inhibited when used in combination with Cu (II). It was due to the interaction of 5-FU with Cu (II). Hence, we propose that during chemoradiotherapy with 5-FU, the endogenous copper is mobilized by 5-FU, leading to the generation of ROS which cause oxidative stress and possibly cancer cell death by apoptosis. PMID- 21618036 TI - Vertical inheritance and bursts of transposition have shaped the evolution of the BS non-LTR retrotransposon in Drosophila. AB - The history of transposable elements over evolutionary time can often be partially reconstructed on the basis of genome analysis. In this study, we identified and extensively characterized the NLTR BS retrotransposon in 12 sequenced Drosophila genomes, by its sequence diversity within and among genomes, its degeneration pattern and its transcriptional activity. We show that the BS element has a variable copy number and patchy distribution within the Drosophila genus, that it is at distinct stages of the evolutionary cycle in the different Drosophila species and that its evolution is characterized by vertical transmission and by bursts of transposition in certain species. PMID- 21618037 TI - Is laparoscopic cholecystectomy safe for lymphangioma of the gallbladder? A complicated case mimicking subhepatic abscess. AB - Lymphangiomas are rare benign neoplasms of the lymphatic tissue generally occurring in the childhood. Cystic lymphangioma of the gallbladder is an extremely rare tumor with only eight cases having been reported in the literature. The aspecific and potentially misleading clinical presentation of these tumors requires complex preoperative imaging in the setting of clinical suspicion to make the correct diagnosis. The treatment of choice is complete excision with negative margins to avoid local recurrence. Their tendency to locally invade the surrounding tissues requires sometimes extended resections. Laparoscopic cholecystectomy can be a questionable choice in this setting; however, the procedures can be performed safely in most cases, although complicated. We report the case of a hemorrhagic cystic lymphangioma of the gallbladder mimicking a subhepatic abscess and operated in emergency with laparoscopic approach. PMID- 21618038 TI - When flavor guides motor control: an effector independence study. AB - Research on multisensory integration during natural tasks has revealed how chemical senses contribute to plan and control movements. An aspect which has yet to be investigated regards whether the motor representations evoked by chemosensory stimuli, once established for a particular movement, can be used to control different effectors. Here, we investigate this issue by asking participants to drink a sip of flavored solution, grasp with the hand a visual target, and then bring it to the mouth, miming the action of biting. Results show that hand and lip apertures were scaled according to the size of the object evoked by the flavor. Maximum hand and lip apertures were greater when the action toward a small visual target (e.g., strawberry) was preceded by a sip of a "large" (e.g., orange) than a "small" (e.g., almond) flavor solution. Conversely, maximum hand and lip apertures were smaller when the action toward a large visual target (e.g., apple) was preceded by the presentation of a "small" (e.g., strawberry) rather than a "large" flavor solution. These findings support previous evidence on the presence of a unique motor plan underlying the act of grasping with-the-hand and with-the-mouth, extending the knowledge of chemosensorimotor transformations to motor equivalence. PMID- 21618039 TI - Subclavian artery perforation complicating coronary angiography. PMID- 21618040 TI - Obstructive intramyocardial haematoma after percutaneous coronary intervention. PMID- 21618041 TI - Paroxysmal mechanical aortic valve prosthesis dysfunction: capturing the right moment. PMID- 21618043 TI - Quantification of cardiorespiratory interactions based on joint symbolic dynamics. AB - Cardiac and respiratory rhythms are highly nonlinear and nonstationary. As a result traditional time-domain techniques are often inadequate to characterize their complex dynamics. In this article, we introduce a novel technique to investigate the interactions between R-R intervals and respiratory phases based on their joint symbolic dynamics. To evaluate the technique, electrocardiograms (ECG) and respiratory signals were recorded in 13 healthy subjects in different body postures during spontaneous and controlled breathing. Herein, the R-R time series were extracted from ECG and respiratory phases were obtained from abdomen impedance belts using the Hilbert transform. Both time series were transformed into ternary symbol vectors based on the changes between two successive R-R intervals or respiratory phases. Subsequently, words of different symbol lengths were formed and the correspondence between the two series of words was determined to quantify the interaction between cardiac and respiratory cycles. To validate our results, respiratory sinus arrhythmia (RSA) was further studied using the phase-averaged characterization of the RSA pattern. The percentage of similarity of the sequence of symbols, between the respective words of the two series determined by joint symbolic dynamics, was significantly reduced in the upright position compared to the supine position (26.4 +/- 4.7 vs. 20.5 +/- 5.4%, p < 0.01). Similarly, RSA was also reduced during upright posture, but the difference was less significant (0.11 +/- 0.02 vs. 0.08 +/- 0.01 s, p < 0.05). In conclusion, joint symbolic dynamics provides a new efficient technique for the analysis of cardiorespiratory interaction that is highly sensitive to the effects of orthostatic challenge. PMID- 21618044 TI - Inelasticity of human carotid atherosclerotic plaque. AB - Little mechanical test data exists regarding the inelastic behavior of atherosclerotic plaques. As a result finite element (FE) models of stenting procedures commonly use hyperelastic material models to describe the soft tissue response thus limiting the accuracy of the model to the expansion stage of stent implantation and leave them unable to predict the lumen gain. In this study, cyclic mechanical tests were performed to characterize the inelastic behavior of fresh human carotid atherosclerotic plaque tissue due to radial compressive loading. Plaques were classified clinically as either mixed (M), calcified (Ca), or echolucent (E). An approximately linear increase in the plastic deformation was observed with increases in the peak applied strain for all plaque types. While calcified plaques generally appeared stiffest, it was observed that the clinical classification of plaques had no significant effect on the magnitude of permanent deformation on unloading. The test data was characterized using a constitutive model that accounts for both permanent deformation and stress softening to describe the compressive plaque behavior on unloading. Material constants are reported for individual plaques as well as mean values for each plaque classification. This data can be considered as a first step in characterizing the inelastic behavior of atherosclerotic plaques and could be used in combination with future mechanical data to improve the predictive capabilities of FE models of angioplasty and stenting procedures particularly in relation to lumen gain. PMID- 21618045 TI - Adult Baby Syndrome and Gender Identity Disorder. PMID- 21618046 TI - Mean age of first sex: do they know what we mean? PMID- 21618047 TI - Nitrifying granules cultivation in a sequencing batch reactor at a low organics to-total nitrogen ratio in wastewater. AB - It is possible to cultivate aerobic granular sludge at a low organic loading rate and organics-to-total nitrogen (COD/N) ratio in wastewater in the reactor with typical geometry (height/diameter = 2.1, superficial air velocity = 6 mm/s). The noted nitrification efficiency was very high (99%). At the highest applied ammonia load (0.3 +/- 0.002 mg NH (4) (+) -N g total suspended solids (TSS)(-1) day(-1), COD/N = 1), the dominating oxidized form of nitrogen was nitrite. Despite a constant aeration in the reactor, denitrification occurred in the structure of granules. Applied molecular techniques allowed the changes in the ammonia-oxidizing bacteria (AOB) community in granular sludge to be tracked. The major factor influencing AOB number and species composition was ammonia load. At the ammonia load of 0.3 +/- 0.002 mg NH (4) (+) -N g TSS(-1) day(-1), a highly diverse AOB community covering bacteria belonging to both the Nitrosospira and Nitrosomonas genera accounted for ca. 40% of the total bacteria in the biomass. PMID- 21618048 TI - Neuroprotective effects of 2-cyclopropylimino-3-methyl-1,3-thiazoline hydrochloride against oxidative stress. AB - Oxidative stress, glutamate excitotoxicity, and inflammation are the important pathological mechanisms in neurodegenerative diseases. Recently, we reported that 2-cyclopropylimino-3-methyl-1,3-thiazoline hydrochloride protects rat glial cells against glutamate-induced excitotoxicity. In this study, we report the effects of 2-cyclopropylimino-3-methyl-1,3-thiazoline hydrochloride on primary cultured cortical astrocytes after exposure to hydrogen peroxide (H2O2). Pretreatment of cells with 2-cyclopropylimino-3-methyl-1,3-thiazoline hydrochloride prior to H2O2 exposure attenuated the H2O2-induced reductions in cell survival and superoxide dismutase, catalase, glutathione, and glutathione peroxidase activities. It also reduced H2O2-induced increases in reactive oxygen species levels, malondialdehyde content, and production of nitric oxide. These effects were all concentration dependent. Our results suggest that 2-cyclopropylimino-3-methyl-1,3-thiazoline hydrochloride protects against oxidative stress. PMID- 21618049 TI - Nitric oxide contributes to hypoxia-reoxygenation-induced P-glycoprotein expression in rat brain endothelial cells. AB - Ischemia-reperfusion leads to increased levels at the blood-brain barrier of the multidrug efflux transporter, P-glycoprotein that provides protection to the brain by limiting access of unwanted substances. This is coincident with the production of nitric oxide. This present study using immortalized rat brain endothelial cells (GPNTs) examines whether following hypoxia-reoxygenation, nitric oxide contributes to the alterations in P-glycoprotein levels. After 6 h of hypoxia, both nitric oxide and reactive oxygen species, detected intracellularly using fluorescent monitoring dyes, were produced in the subsequent reoxygenation phase coincident with increased P-glycoprotein. The evidence that nitric oxide can directly affect P-glycoprotein expression was sought by applying S-nitroso-N-acetyl-DL: -penicillamine that as shown increased the nitric oxide generation. Sodium nitroprusside, though more effective at increasing P-glycoprotein expression, appeared to produce different reactive species. Real time RT-PCR analysis revealed the predominant form of nitric oxide synthase in these cells to be endothelial, inhibition of which partially prevented the increase in P-glycoprotein during reoxygenation. These data indicate that the production of nitric oxide by endothelial nitric oxide synthase during reoxygenation can influence P-glycoprotein expression in cells of the blood-rat brain barrier, highlighting another route by which nitric oxide may protect the brain. PMID- 21618050 TI - Ogataea saltuana sp. nov., a novel methanol-assimilating yeast species. AB - Four ascosporulating strains of an undescribed methanol-assimilating yeast species were isolated from forest habitats in Hungary. Three were recovered from rotten wood and one from leaves of a sessile oak (Quercus petraea). An additional isolate of the undescribed species sharing similar phenotypic characters with the above-noted strains was recovered from the gut of an unidentified beetle collected from under the bark of a coniferous tree in Bulgaria. A closely related, but somewhat divergent strain was recovered from insect frass in a Ponderosa pine (Pinus ponderosa) collected in New Mexico, USA. Analysis of the D1/D2 sequences of the LSU rRNA gene placed the new species in the Ogataea clade. The ITS and the D1/D2 LSU sequences of the rRNA gene repeats were compared for the above-noted strains and that of the type strain of Ogataea zsoltii, the closest neighbour among currently recognized Ogataea species. Their relatedness was investigated by parsimony network analysis as well. As a result of the sequence analysis, it was concluded that the six strains isolated from tree associated habitats represent a single new yeast species. Ogataea saltuana sp. nov. is proposed to accommodate these strains. The type strain NCAIM Y.01833(T) (CBS 10795(T), NRRL Y-48448(T)) was recovered from rotten wood of Scotch pine (Pinus silvestris) in Hungary. The GenBank accession number for the D1/D2 domain nuclear large subunit rRNA gene sequence of strain NCAIM Y.01833(T) (CBS 10795(T), NRRL Y-48448(T)) is EU327033. The MycoBank number of the new species is MB 519966. PMID- 21618051 TI - A re-audit of prevention of venous thromboembolic disease in medical and surgical hospital patients in a single university teaching hospital. PMID- 21618052 TI - Soothing a sore throat: the efficacy and safety of steroids in acute pharyngitis. AB - BACKGROUND: Acute pharyngitis is a common presentation to Emergency Departments (EDs) and although most patients experience mild-to-moderate discomfort, severe pharyngitis can be incapacitating and painful. There have been recent trends in prescribing steroids to provide pain relief for patients with acute pharyngitis. AIM: To examine the evidence for using steroids in adult patients with acute pharyngitis attending Emergency Departments. METHODS: A comprehensive literature search was performed to identify randomised controlled trials, systematic reviews and meta-analyses. The primary outcome measure assessed was pain relief, while safety of steroids was chosen as a secondary outcome measure. RESULTS: Six papers met the inclusion criteria--five randomised controlled clinical trials and one systematic review (adult and paediatric patients in a variety of clinical settings). Mean times to onset of pain relief were shorter in patients receiving steroids (8.1 vs. 19.9 h, 6.3 vs. 11.3 h, 6.3 vs. 12.4 h). A single study examining reduction in VAS pain scores found a statistically significant improvement in patients given steroids (PO: 4.2 +/- 2.3, IM: 3.8 +/- 2.3 vs placebo: 2.0). Marvez-Valls et al. reported shorter times to complete pain relief (43 vs. 36 h) but this and other studies failed to demonstrate a statistically significant reduction in the pain scores at 24 h. Frequency and nature of side effect reporting was inconsistent throughout the studies with no trials explicitly powered to detect clinically important adverse events. CONCLUSION: Steroids may provide a useful adjunct in the management of acute pharyngitis, but there is currently insufficient evidence to endorse routine use. PMID- 21618053 TI - A comprehensive workflow for general-purpose neural modeling with highly configurable neuromorphic hardware systems. AB - In this article, we present a methodological framework that meets novel requirements emerging from upcoming types of accelerated and highly configurable neuromorphic hardware systems. We describe in detail a device with 45 million programmable and dynamic synapses that is currently under development, and we sketch the conceptual challenges that arise from taking this platform into operation. More specifically, we aim at the establishment of this neuromorphic system as a flexible and neuroscientifically valuable modeling tool that can be used by non-hardware experts. We consider various functional aspects to be crucial for this purpose, and we introduce a consistent workflow with detailed descriptions of all involved modules that implement the suggested steps: The integration of the hardware interface into the simulator-independent model description language PyNN; a fully automated translation between the PyNN domain and appropriate hardware configurations; an executable specification of the future neuromorphic system that can be seamlessly integrated into this biology-to hardware mapping process as a test bench for all software layers and possible hardware design modifications; an evaluation scheme that deploys models from a dedicated benchmark library, compares the results generated by virtual or prototype hardware devices with reference software simulations and analyzes the differences. The integration of these components into one hardware-software workflow provides an ecosystem for ongoing preparative studies that support the hardware design process and represents the basis for the maturity of the model-to hardware mapping software. The functionality and flexibility of the latter is proven with a variety of experimental results. PMID- 21618054 TI - Biplane correlation imaging: a feasibility study based on phantom and human data. AB - The objective of this study was to implement and evaluate the performance of a biplane correlation imaging (BCI) technique aimed to reduce the effect of anatomic noise and improve the detection of lung nodules in chest radiographs. Seventy-one low-dose posterior-anterior images were acquired from an anthropomorphic chest phantom with 0.28 degrees angular separations over a range of +/-10 degrees along the vertical axis within an 11 s interval. Similar data were acquired from 19 human subjects with institutional review board approval and informed consent. The data were incorporated into a computer-aided detection (CAD) algorithm in which suspect lesions were identified by examining the geometrical correlation of the detected signals that remained relatively constant against variable anatomic backgrounds. The data were analyzed to determine the effect of angular separation, and the overall sensitivity and false-positives for lung nodule detection. The best performance was achieved for angular separations of the projection pairs greater than 5 degrees . Within that range, the technique provided an order of magnitude decrease in the number of false-positive reports when compared with CAD analysis of single-view images. Overall, the technique yielded ~1.1 false-positive per patient with an average sensitivity of 75%. The results indicated that the incorporation of angular information can offer a reduction in the number of false-positives without a notable reduction in sensitivity. The findings suggest that the BCI technique has the potential for clinical implementation as a cost-effective technique to improve the detection of subtle lung nodules with lowered rate of false-positives. PMID- 21618055 TI - Optimizing bioimpedance measurement configuration for dual-gated nuclear medicine imaging: a sensitivity study. AB - Motion artefacts due to respiration and cardiac contractions may deteriorate the quality of nuclear medicine imaging leading to incorrect diagnosis and inadequate treatment. Motion artefacts can be minimized by simultaneous respiratory and cardiac gating, dual-gating. Currently, only cardiac gating is often performed. In this study, an optimized bioimpedance measurement configuration was determined for simultaneous respiratory and cardiac gating signal acquisition. The optimized configuration was located on anterolateral upper thorax based on sensitivity simulations utilizing a simplified thorax model. The validity of the optimized configuration was studied with six healthy volunteers. In the peak-to-peak and frequency content analyses the optimized configuration showed consistently higher peak-to-peak values and frequency content than other studied measurement configurations. This study indicates that the bioimpedance method has potential for the dual-gating in nuclear medicine imaging. The method would minimize the need of additional equipment, is easy for the technologists to use and comfortable for the patients. PMID- 21618056 TI - Retrospective analysis of seventy-one patients with neuroendocrine tumor and review of the literature. AB - Neuroendocrine tumors (NET) are rare, but their incidence is gradually increasing. In this study, demographical and tumor characteristics, treatment modalities, responses, and survival rates were evaluated in the patients with NET. Seventy-one patients with NET from 3 tertiary care centers evaluated retrospectively. Overall survival (OS), progression-free survival (PFS), and disease-free survival rates were estimated by Kaplan-Meier Method. Male/female ratio was 0.86 (33/38). Median age was 52 years. Rates for family cancer history and goiter/thyroiditis were 22.4 and 17.8%, respectively. The most common primary site was lung (22.5%), in parallel with the literature, and 31% had the large cell neuroendocrine carcinoma histology. The second most common site was stomach. Carcinoid syndrome rate was found to be 30.6%. Half of the patients were in early stage at diagnosis. Surgical resection rate was 64.7, and 45% of the patient received chemotherapy (CT), 22% received radiotherapy. Seventy-six percent of resected patients had local disease. Thirty-two patients received CT for palliation or concurrent with radiotherapy or in adjuvant setting. Platin/etoposide combination was the most commonly used chemotherapy regimen. Chemotherapy response rate was 35.7%. Five patients had received somatostatin analogue. Radiotherapy was used in adjuvant setting in one-third of the patients. Median OS was 66 months, and median PFS was 30 months. Female gender and fifth decade seem to have higher risk. History for family cancer and goiter/thyroiditis was high in the patients with NET, though there is no data about an association between NET and thyroid disorders in the literature. PMID- 21618057 TI - Acute transient encephalopathy after weekly paclitaxel infusion. AB - Paclitaxel is highly active against a variety of solid tumors including breast lung, ovarian and head and neck cancer. Although peripheral neurotoxicity is well known side effect, central nervous system (CNS) toxicity-related standard dose of paclitaxel is extremely uncommon, because paclitaxel dose not cross the blood brain barrier and is not detectable in the cerebrospinal fluid. We present a patient with advanced stage breast carcinoma who developed acute and spontaneous resolving encephalopathy after weekly dose of paclitaxel. The patient did not have brain metastasis, or prior whole-brain irradiation, or any type of neurosurgery. Radiological imaging studies showed no abnormalities. CNS toxicity of paclitaxel should be kept in mind in patients without a previous history of brain metastasis or brain irradiation and even with low weekly doses. PMID- 21618058 TI - Rat pancreatic level of cystathionine gamma-lyase is regulated by glucose level via specificity protein 1 (SP1) phosphorylation. AB - AIMS/HYPOTHESIS: Cystathionine gamma-lyase (CSE) catalyses the endogenous production of hydrogen sulphide (H(2)S) in pancreatic beta cells, and H(2)S has been shown to inhibit insulin release from these cells. As altered pancreatic H(2)S production modulated by glucose has been previously shown, we hypothesised that the Cse gene could be regulated by glucose level in insulin-secreting cells. METHODS: The effects of glucose on CSE protein level and mRNA level were analysed in INS-1E cells. Glucose effect on Cse promoter activity was tested by constructing a proximal Cse promoter vector including specificity protein 1 (Sp1) consensus sequence. RESULTS: High glucose (20 mmol/l) inhibited H(2)S production in INS-1E cells and freshly isolated rat pancreatic islets. Cse mRNA expression, CSE activity and protein abundance were also profoundly reduced by high glucose. The involvement of SP1 in basal and high-glucose-regulated CSE production was demonstrated. Sp1-knockdown abolished a large portion of CSE production at basal glucose. Phosphorylation of SP1 stimulated by high glucose was inhibited by p38 mitogen-activated protein kinase (MAPK) inhibitors SB203580 and SB202190. After blocking p38 MAPK phosphorylation, the inhibitive effects of high glucose on CSE protein production and promoter activity in INS-1E cells were also virtually abolished. CONCLUSIONS/INTERPRETATION: Glucose stimulates the phosphorylation of SP1 via p38 MAPK activation, which leads to decreased Cse promoter activity and subsequent downregulation of Cse gene expression. Inhibited H(2)S production through glucose-mediated CSE activity and production alterations may be involved in the fine control of glucose-induced insulin secretion. PMID- 21618059 TI - Development of a complex parent-metabolite joint population pharmacokinetic model. AB - This study aimed to develop a joint population pharmacokinetic model for an antipsychotic agent in development (S33138) and its active metabolite (S35424) produced by reversible metabolism. Because such a model leads to identifiability problems and numerical difficulties, the model building was performed using the FOCE-I and the Stochastic Approximation Expectation Maximization (SAEM) estimation algorithms in NONMEM and MONOLIX, respectively. Four different structural models were compared based on Bayesian information criteria. Models were first written as ordinary differential equations systems and then in closed form (CF) to facilitate further analyses. The impact of polymorphisms on genes coding for the CYP2C19 and CYP2D6 enzymes, respectively involved in the parent drug and the metabolite elimination were investigated using permutation Wald test. The parent drug and metabolite plasma concentrations of 101 patients were analyzed on two occasions after 4 and 8 weeks of treatment at 1, 3, 6, and 24 h following daily oral administration. All configurations led to a two compartment model with back-transformation of the metabolite into the parent drug and a first pass effect. The elimination clearance of the metabolite through other processes than back-transformation was decreased by 35% [9-53%] in CYP2D6 poor metabolizer. Permutation tests were performed to ensure the robustness of the analysis, using SAEM and CF. In conclusion, we developed a complex joint pharmacokinetic model adequately predicting the impact of CYP2D6 polymorphisms on the parent drug and its metabolite concentrations through the back-transformation mechanism. PMID- 21618060 TI - Practice guidelines for communicating a prenatal or postnatal diagnosis of Down syndrome: recommendations of the national society of genetic counselors. AB - Down syndrome is one of the most common conditions encountered in the genetics clinic. Due to improvements in healthcare, educational opportunities, and community inclusion over the past 30 years, the life expectancy and quality of life for individuals with Down syndrome have significantly improved. As prenatal screening and diagnostic techniques have become more enhanced and widely available, genetic counselors can expect to frequently provide information and support following a new diagnosis of Down syndrome. This guideline was written for genetic counselors and other healthcare providers regarding the communication of a diagnosis of Down syndrome to ensure that families are consistently given up to-date and balanced information about the condition, delivered in a supportive and respectful manner. PMID- 21618061 TI - Vacuum phenomenon: prevalence and appearance in the knee with 3 T magnetic resonance imaging. AB - OBJECTIVES: To determine the prevalence of vacuum phenomenon (VP) in the knee on magnetic resonance (MR) images, describe the imaging features that characterize VP, and assess how often VP mimics pathological knee lesions. MATERIALS AND METHODS: Consecutive knee MR studies performed on a 3 T MR system over a 9-month period were retrospectively reviewed by one radiologist who then selected studies with findings potentially indicating VP. Three experienced musculoskeletal radiologists reviewed these cases in consensus to confirm the presence of VP and to assess the shape, size, and signal of VP; the presence of magnetic susceptibility artifacts; and the ability of MR sequences to show VP. RESULTS: A total of 914 consecutive exams from 875 patients (524 men; mean age, 35 years) were reviewed. Vacuum phenomenon was found in 12 patients (prevalence 1.3%). In six (50%) patients, VP mimicked a meniscal tear, with four cases simulating a torn medial discoid meniscus. The VP signal was not easily differentiated from meniscal signal on most sequences in most cases (9/12). Gradient-recalled echo (GRE) localizer images proved most definitive, with 3D SPACE images the next most effective. Fast spin echo (FSE) images were only occasionally able to differentiate VP from meniscus. CONCLUSION: Rarely recognized on MR, VP can mimic meniscal pathology, potentially leading to inappropriate surgery. Because differentiation of VP from the meniscus is challenging on FSE at 3 T, radiologists should become familiar with the appearance of VP and review GRE localizer or 3D images carefully to avoid misinterpretation. PMID- 21618062 TI - Potent inhibition of protein tyrosine phosphatases by copper complexes with multi benzimidazole derivatives. AB - A series of copper complexes with multi-benzimidazole derivatives, including mono and di-nuclear, were synthesized and characterized by Fourier transform IR spectroscopy, UV-Vis spectroscopy, elemental analysis, electrospray ionization mass spectrometry. The speciation of Cu/NTB in aqueous solution was investigated by potentiometric pH titrations. Their inhibitory effects against human protein tyrosine phosphatase 1B (PTP1B), T-cell protein tyrosine phosphatase (TCPTP), megakaryocyte protein tyrosine phosphatase 2 (PTP-MEG2), srchomology phosphatase 1 (SHP-1) and srchomology phosphatase 2 (SHP-2) were evaluated in vitro. The five copper complexes exhibit potent inhibition against PTP1B, TCPTP and PTP-MEG2 with almost same inhibitory effects with IC(50) at submicro molar level and about tenfold weaker inhibition versus SHP-1, but almost no inhibition against SHP-2. Kinetic analysis indicates that they are reversible competitive inhibitors of PTP1B. Fluorescence study on the interaction between PTP1B and complex 2 or 4 suggests that the complexes bind to PTP1B with the formation of a 1:1 complex. The binding constant are about 1.14 * 10(6) and 1.87 * 10(6) M(-1) at 310 K for 2 and 4, respectively. PMID- 21618063 TI - A 5-year experience with laparoscopic adjustable gastric banding--focus on outcomes, complications, and their management. AB - BACKGROUND: Laparoscopic adjustable gastric banding (LAGB) remains the most popular surgical modality for obesity management in Europe. The aim of this publication is to present a 5-year experience in obesity treatment with LAGB operation with the assessment of outcomes, frequency of complications, and their management. Management of the band-related complications is crucial for continuous obesity treatments, despite the fact of initial failure, allowing further excess weight loss in patients with morbid obesity. METHODS: One hundred sixty patients underwent the LAGB procedure with standard pars flaccida technique during the years 2005-2009. A retrospective analysis of the data was performed; chi-squared test and Student's t test at the level of significance of p < 0.05 were used. Information on reoperations was gathered from hospital case notes. RESULTS: In the presented group, the mean body mass index (BMI) was 48.13 kg/m(2) (33.46-83.04 kg/m(2); standard deviation [SD] +/-8.45). Of the patients, 36.2% had super morbid obesity with BMI >50 kg/m(2). The mean observation period reached 549 days (31-2,026 days; SD +/-390.1), with the mean number of control visits of 4.2 (1-12). The mean percentage of excess weight loss during the observation period was 34% (from -9.9% to 85.1%; SD +/-20.6), with the mean body mass reduction of 24.4 kg. Complications appeared in 30 patients (20.1%). Twenty four patients (16.1%) required reoperation. There were no mortalities recorded. CONCLUSIONS: The mean operative time of 59 min was relatively short. Morbidity and mortality rates were comparable to many published series. Failure or complications of LAGB did not stop the obesity treatment. Most of the band related complications occurred late and could be provided for laparoscopically. PMID- 21618064 TI - Decreased serum betaine concentrations in patients after bariatric surgery. AB - Bariatric surgery significantly reduces the risk of cardiovascular diseases but has no effects on hyperhomocysteinemia, the risk factor for atherogenesis. We hypothesize that the decrease in serum betaine (involved in homocysteine metabolism) concentrations, after bariatric surgery, impairs conversion of homocysteine to methionine, leading to hyperhomocysteinemia. If this is true, it may be desirable to supply patients after bariatric surgery with betaine. Serum betaine and homocysteine concentrations were measured by liquid chromatography/mass spectrometry, in 16 obese patients, before and 6 months after bariatric surgery. Ten healthy individuals with normal body mass index served as controls. Serum betaine concentrations decreased to the values lower than in controls after bariatric surgery, whereas serum homocysteine concentrations remained elevated. In patients supplemented with B(12) and folate, no effect of bariatric surgery on serum concentrations of vitamins involved in homocysteine metabolism was observed. These results suggest that betaine deficit could be responsible for maintenance of hyperhomocysteinemia after bariatric surgery. We postulate that supplementation with betaine could be of therapeutic value for the treatment of hyperhomocysteinemia after bariatric surgery. PMID- 21618065 TI - Predictive factors of participation in postsecondary education for high school leavers with autism. AB - This exploratory study was designed to identify the factors predictive of participation in postsecondary education for high school leavers with autism. A secondary data analysis of the National Longitudinal Transition Study 2 (NLTS2) data was performed for this study. Potential predictors of participation in postsecondary education were assessed using a backward logistic regression analysis. This study found that the high school's primary post-high school goal for the student, parental expectations, high school type, annual household income, and academic performance were significant predictors of participation in postsecondary education. The findings of this current study may provide critical information for parents of children with autism as well as educators and professionals who work with students with autism. PMID- 21618066 TI - Larger IOR effects following forget than following remember instructions depend on exogenous attentional withdrawal and target localization. AB - When words are onset in the visual periphery, inhibition of return (IOR) for a subsequent target is larger when those words receive an intervening forget instruction than when they receive a remember instruction Taylor (Quarterly Journal of Experimental Psychology, 58A, 613-629, 2005). The present study manipulated the allocation of endogenous and exogenous attention to assess the source of the forget > remember IOR difference. We determined that the forget > remember IOR difference likely arises from the differential withdrawal of exogenous-rather than endogenous-attention. Furthermore, this forget > remember IOR difference occurs only when a spatially compatible localization response is required; it does not occur when a simple detection response or a perceptual discrimination is required. This suggests that the forget > remember difference in the magnitude of IOR is not due to differences in perceptual/attentional processing. Instead, an instruction to remember or forget biases spatial responses in accordance with whether a location has previously contained relevant or irrelevant information. We suggest that directed forgetting in an item-method paradigm is not accomplished by changes in attention; rather, the changes in attention are coincident with changes in memory and may serve to bias later responses away from a source of unreliable information. PMID- 21618067 TI - Dose-dependent influence of sevoflurane anesthesia on neuronal survival and cognitive outcome after transient forebrain ischemia in Sprague-Dawley rats. AB - BACKGROUND: Volatile anesthetics reduce postischemic neurohistopathological injury and improve neurological outcome in various animal models. However, the isoflurane concentrations above 1 minimum alveolar concentration (MAC) have been associated with reduced neuronal survival and impaired functional outcome. The aim of this study was to evaluate if 1.8 MAC sevoflurane alters postischemic neuronal survival and neurologic outcome compared with 0.45 MAC sevoflurane. METHODS: In this study, 20 fasted male Sprague-Dawley rats were randomly assigned to treatment groups with 1 or 4 vol.% sevoflurane end-tidal concentration. Cerebral ischemia was induced by bilateral carotid artery occlusion and hemorrhagic hypotension (BCAO). The cognitive outcome was assessed after 7 days using the object recognition test. Animals were then re-anesthetized and brains were removed for neurohistopathological analysis of the hippocampus (CA1) and cortex using hematoxylin-eosin staining. RESULTS: Physiologic parameters were not different between both the treatment groups. The number of viable neurons (median [Q1, Q3]) in the CA1 region on postischemic day 7 was increased after high-dose sevoflurane compared with low-dose sevoflurane (1645 [453, 1825] vs. 3222 [2920, 3993] neurons/ROI, P < 0.05). Results of the object recognition test were not different between both the treatment groups. CONCLUSIONS: Postischemic neuronal survival was increased with 1.8 MAC compared with 0.45 MAC sevoflurane. Therefore, experimental models of cerebral ischemia should account for neuroprotective effects of sevoflurane with increasing concentrations. To ensure minimal interference of sevoflurane on neuronal survival, a low inspired concentration should be used and fluctuations in the depth of anesthesia should be limited. PMID- 21618068 TI - Understanding the "Russian mortality paradox" in Central Asia: evidence from Kyrgyzstan. AB - In the former Soviet republics of central Asia, ethnic Russians have exhibited higher adult mortality than native ethnic groups (e.g., Kazakh, Kyrgyz, Uzbek) in spite of the higher socioeconomic status of ethnic Russians. The mortality disadvantage of ethnic Russians at adult ages appears to have even increased since the breakup of the Soviet Union. The most common explanation for this "Russian mortality paradox," is that deaths are better reported among ethnic Russians. In this study, we use detailed mortality data from Kyrgyzstan between 1959 and 1999 to evaluate various explanations for the Russian mortality paradox: data artifacts, migration effects, and cultural effects. We find that the most plausible explanation is the cultural hypothesis because the personal behaviors that appear to generate a large part of the observed mortality differences (alcohol consumption, in particular) seem to be closely tied to cultural practices. We examine the implications of this finding for understanding the health crisis in post-Soviet states. PMID- 21618070 TI - Specific effects of a calorie-based intervention on stair climbing in overweight commuters. AB - BACKGROUND: Point-of-choice prompts consistently increase stair climbing; a greater increase in overweight than normal weight individuals was reported in a multi-component worksite campaign. PURPOSE: The purpose of this study is to investigate effects of a multi-component campaign, on stair climbing, in a public access setting. METHODS: In an interrupted-time-series-design, baseline observations (2 weeks) preceded a 2-week point-of-choice prompt. An additional message, positioned at the top of the climb for a further 6-week period, summarised the calorific consequences of a single ascent. Inconspicuous observers recorded traveller's methods of ascent, coded by sex and weight status, twice a week between 08:00 and 09:59. RESULTS: At baseline, the overweight chose stairs less than normal weight individuals. The multi-component campaign targeting weight control reversed this bias, increasing stair climbing only in overweight individuals. CONCLUSIONS: The specificity of the effect confirms the appeal of this lifestyle activity for the overweight. The discussion focuses on how intentions to control weight may be converted into behaviour. PMID- 21618071 TI - Plant-pathogen interactions: what microarray tells about it? AB - Plant defense responses are mediated by elementary regulatory proteins that affect expression of thousands of genes. Over the last decade, microarray technology has played a key role in deciphering the underlying networks of gene regulation in plants that lead to a wide variety of defence responses. Microarray is an important tool to quantify and profile the expression of thousands of genes simultaneously, with two main aims: (1) gene discovery and (2) global expression profiling. Several microarray technologies are currently in use; most include a glass slide platform with spotted cDNA or oligonucleotides. Till date, microarray technology has been used in the identification of regulatory genes, end-point defence genes, to understand the signal transduction processes underlying disease resistance and its intimate links to other physiological pathways. Microarray technology can be used for in-depth, simultaneous profiling of host/pathogen genes as the disease progresses from infection to resistance/susceptibility at different developmental stages of the host, which can be done in different environments, for clearer understanding of the processes involved. A thorough knowledge of plant disease resistance using successful combination of microarray and other high throughput techniques, as well as biochemical, genetic, and cell biological experiments is needed for practical application to secure and stabilize yield of many crop plants. This review starts with a brief introduction to microarray technology, followed by the basics of plant-pathogen interaction, the use of DNA microarrays over the last decade to unravel the mysteries of plant pathogen interaction, and ends with the future prospects of this technology. PMID- 21618072 TI - Population fragmentation causes randomly fixed genotypes in populations of Arabidopsis kamchatica in the Japanese Archipelago. AB - Populations of arctic alpine plants likely disappeared and re-colonised several times at the southern edge of their distributions during glacial and interglacial cycles throughout the Quaternary. Range shift and population fragmentation after a glacial period would affect the genetic structure of such plants in southernmost populations. We aimed to elucidate how climatic oscillations influenced the population subsistence of alpine plants in the Japanese Archipelago as one of the southernmost populations, by inferring the genetic structure of Arabidopsis kamchatica subsp. kamchatica and the intraspecific littoral taxon, subsp. kawasakiana. We identified genotypes based on the haplotypes of five nuclear genes and two chloroplast DNA spacers for 164 individuals from 24 populations. Most populations harboured only one private genotype, whereas few polymorphisms were found in each population. Two genetic genealogies were found, suggesting that northern Japanese populations of alpine subsp. kamchatica, subsp. kawasakiana and the northerly subsp. kamchatica in eastern Russia and Alaska clustered and differentiated from populations in central Honshu, western Japan and Taiwan. During climatic oscillations, the genetic structure of extant southernmost populations would have been shaped by strong genetic drift under population fragmentation and randomly fixed to a single genotype among their ancestral polymorphisms. PMID- 21618073 TI - Late tracheostomy tube decannulation by progression of a laryngeal tumour: an approach for airway control. PMID- 21618074 TI - Rates of early intervention referral and significant developmental delay, by birthweight and gestational age. AB - Though correlated, birthweight (BW) and gestational age (GA) have independent effects on cognitive and neurological outcomes. Jurisdictions vary in their inclusion of these two characteristics in their list of established conditions for automatic eligibility for Early Intervention (EI) services, which may lead them to miss important high-risk groups. We evaluated the relationship between BW GA combinations and both EI referral rates and risk of EI-diagnosed significant developmental delay in a population of New York City (NYC) births. We linked birth certificates of children born in NYC to resident mothers during 1999-2001 and surviving the first 28 days of life (n = 339,522) to EI administrative data. We calculated EI referral rates for various BW-GA categories, and used a logistic model to directly estimate the predicted risk of delay. EI referral rates of over 50% were observed in children born <1,250 g and those born <30 weeks and 1,250 1,499 g. Additionally, more than one in two children born either less than 1,250 g or <30 weeks and 1,250-1,499 g were predicted to be diagnosed with a developmental delay, compared with almost one-tenth among those born >2,500 g and 39+ weeks. A BW threshold of <1,250 g would identify children with the highest risk of delay; GA as an additional criterion would prevent overlooking high-risk children born <30 weeks but at higher birthweights. Physicians should monitor children with high-risk birth characteristics and refer them, if appropriate, for formal evaluation. EI programs may use these findings to guide determination of automatic eligibility criteria. PMID- 21618075 TI - Successful therapy with tonsillectomy for severe ISKDC grade VI Henoch-Schonlein purpura nephritis and persistent nephrotic syndrome. AB - Henoch-Schonlein purpura (HSP) is a systemic disorder characterized by leukocytoclastic vasculitis involving the capillaries and the deposition of IgA immune complexes. Renal involvement is the principal cause of morbidity and mortality in children with HSP. We report here a 13-year-old girl with Henoch Schonlein purpura nephritis (HSPN) of International Study of Kidney Disease in Children (ISKDC) grade VI and persistent nephrotic syndrome despite receiving conventional therapy, such as prednisolone, methylprednisolone and urokinase pulse therapy and plasmapheresis (PP). The patient was treated with tonsillectomy, which subsequently decreased proteinuria, induced the disappearance of microscopic hematuria, and improved renal pathological findings. A regimen of methylprednisolone and urokinase pulse therapy plus PP with tonsillectomy may be an effective and useful therapy for some children with severe HSPN children of ISKDC grade VI and persistent nephrotic syndrome. PMID- 21618076 TI - Anaerobic clavicular osteomyelitis following colonoscopy in a hemodialysis patient. AB - Patients on dialysis are immunocompromised and are therefore susceptible to both common and unusual infectious complications. These infections are often related to their dialysis access but even routine diagnostic tests unrelated to dialysis can also lead to rare adverse events. We present an unusual case of clavicular osteomyelitis from Bacteroides fragilis in a patient on maintenance hemodialysis following colonoscopy. The risk factors for this unusual site of infection, the incidence and guidelines for prophylactic antibiotic administration are discussed here. PMID- 21618077 TI - The efficacy and safety of etanercept in patients with rheumatoid arthritis and spondyloarthropathy on hemodialysis. AB - We aimed to evaluate the efficacy and safety of long-term use of etanercept therapy in patients with spondyloarthropathy (SpA) and rheumatoid arthritis (RA) on hemodialysis (HD). Selected RA or SpA patients treated with etanercept under HD were retrospectively evaluated. Etanercept-related adverse events were closely recorded for all patients. At the follow-up, erythrocyte sedimentation rate and C reactive protein levels were monitored. Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) for SpA patients and Disease Activity Score (DAS28) for RA patients were measured at every 3 or 6 months. In total five end-stage renal disease (ESRD) patients were enrolled to the study. The causes of ESRD in the study subjects were amyloidosis (n = 2), analgesic nephropathy (n = 2), and nephrolithiasis (n = 1). Three were diagnosed as SpA and two were RA. All patients used etanercept. The median age was 39 years (range 22-72 years). The median disease duration was 12 years (range 2-20 years). The median follow-up after etanercept therapy was 18 months (range 5-33 months). DAS28 score decreased after the treatment and did not increase during follow-up in RA patients. BASDAI score decreased after the treatment during follow-up in three patients with SpA. At the follow-up, only one patient was diagnosed with septic arthritis. As a result of our study, etanercept treatment in RA and SpA patients on HD seems to be safe, well tolerated, and effective in most of the patients. Above all, due to impaired host defense in patients with ESRD, enhanced risk of infections should be kept in mind during follow-up period and larger trials are needed to prove the safety of etanercept in HD patients. PMID- 21618078 TI - No effect of physiotherapy on the serum levels of adipocytokines in patients with ankylosing spondylitis. AB - To investigate the effect of intensive physiotherapy on disease activity and serum levels of adipocytokines in patients with ankylosing spondylitis (AS). Twenty-six patients with AS were included in this study. Intensive physiotherapy was performed twice a week for a period of 3 months. The Bath AS Disease Activity Index (BASDAI) and the Bath AS Functional Index (BASFI) were assessed at inclusion and after 3 months. Leptin, adiponectin, resistin and visfatin serum levels were analysed by ELISA assays. Patients had mild to moderate disease activity. Baseline levels of adipocytokines did not correlate with indicators of disease activity, functional status or acute-phase reactants. After the 3 months of intensive physiotherapy, BASDAI significantly decreased from 2.98 to 1.8 (p = 0.01) and BASFI improved from 2.31 to 1.37 (p = 0.05), while there were no changes in serum levels of CRP, ESR and adipocytokines. In addition, baseline levels of adipocytokines did not predict the change of disease activity or functional ability. Intensive physiotherapy effectively reduces all clinical measures of disease activity, but it is not associated with a significant change in acute-phase reactants or serum levels of adipocytokines. PMID- 21618079 TI - Disruption of endocytic pathway regulatory genes activates autophagy in C. elegans. AB - Autophagy and endocytic pathway are highly regulated catabolic processes. Both processes are crucial for cell growth, development, differentiation, disease and homeostasis and exhibit membrane rearrangement for their function. Autophagy and endocytic pathway represent branches of the lysosomal digestive system, autophagy being responsible for degradation of cytoplasmic components and endocytic pathway for degradation of exogenous substances. Here we report that autophagy is activated when endocytic pathway regulatory genes such as rab-5 and rabx-5 are disrupted. Defects in the ubiquitin binding domain of RABX-5 are critical in activating autophagy. We also observed that the elevated autophagy level does not contribute to lifespan extension of rabx-5 mutant. Our results suggest that autophagy may compensate for the endocytic pathway when regulatory genes for the endocytic pathway malfunction, providing a case of complementation between two functionally related cellular processes. PMID- 21618080 TI - Trafficking of stem cells. AB - Stem cells undergo regulated trafficking from the developmental stages to the adulthood. Stem cell migration is critical to organize developing organs and likely contributes postnatally to tissue regeneration. Here, we review the molecular mechanisms underlying migration of hematopoietic stem cells, neural stem cells, and primordial germ cells, revealing common operative pathways. PMID- 21618081 TI - Migratory strategies of normal and malignant stem cells. AB - The regulated migration of stem cells is critical for organogenesis during development and for tissue -homeostasis and repair during adulthood. Human bone marrow (BM) represents an accessible reservoir containing regenerative cell types from hematopoietic, endothelial, and mesenchymal-stromal lineages that together coordinate hematopoiesis and promote the repair of damaged vasculature and tissues throughout the body. Thus, a detailed understanding of lineage-specific stem cell mobilization, homing, and subsequent engraftment in areas of injury or disease is of critical importance to the rational development of novel cell mediated regenerative therapies. Stem cell trafficking via the circulation from site of origin to peripheral tissues requires fundamental molecular pathways governing (1) niche-specific deadhesion of progenitor cells; (2) chemoattraction to guide progenitor cell homing; and (3) interstitial navigation and adhesion/retention of recruited progenitor cells. This overview chapter summarizes the diversity of migratory strategies employed by hematopoietic, endothelial, and mesenchymal-stromal progenitor cells during repair and regeneration after tissue damage. Further elucidation of stem cell homing and migration pathways will allow greater application of stem cells for targeted cell therapy and/or drug delivery for tissue repair. Strikingly similar migratory mechanisms appear to govern the in vivo migration of recently characterized cancer stem cells (CSC) in leukemias and solid tumors, indicating that conserved principles of stem cell migration and niche specificity will provide new information to target CSC in anticancer therapy. PMID- 21618082 TI - Hematopoietic stem cell characterization and isolation. AB - Hematopoietic stem cells (HSCs) are defined by the capabilities of multi-lineage differentiation and long-term self-renewal. Both these characteristics contribute to maintain the homeostasis of the system and allow the restoration of hematopoiesis after insults, such as infections or therapeutic ablation. Reconstitution after lethal irradiation strictly depends on a third, fundamental property of HSCs: the capability to migrate under the influence of specific chemokines. Directed by a chemotactic compass, after transplant HSCs find their way to the bone marrow, where they eventually home and engraft. HSCs represent a rare population that primarily resides in the bone marrow with an estimated frequency of 0.01% of total nucleated cells. Separating HSCs from differentiated cells that reside in the bone marrow has been the focus of intense investigation for years. In this chapter, we will describe in detail the strategy routinely used by our laboratory to purify murine HSCs, by exploiting their antigenic phenotype (KSL), combined with the physiological capability to efficiently efflux the vital dye Hoechst 33342, generating the so-called Side Population, or SP. PMID- 21618083 TI - Isolation and characterization of adult neural stem cells. AB - It has been thought for a long time that the adult brain is incapable of generating new neurons, or that neurons cannot be added to its complex circuitry. However, recent technology has resulted in an explosion of research demonstrating that neurogenesis, or the birth of new neurons from adult stem cells constitutively occurs in two specific regions of the mammalian brain; namely the subventricular zone and hippocampal dentate gyrus. Adult CNS stem cells exhibit three main characteristics: (1) they are "self-renewing," i.e., they possess a theoretically unlimited ability to produce progeny indistinguishable from themselves, (2) they are proliferative (undergoing mitosis) and (3) they are multipotent for the different neuroectodermal lineages of the CNS, including the different neuronal, and glial subtypes. CNS stem cells and all progenitor cell types are broadly termed "precursors." In this chapter, we describe methods to identify, isolate and experimentally manipulate stem cells of the adult brain. We outline how to prepare a precursor cell culture from naive brain tissue and how to test the "stemness" potential of different cell types present in that culture, which is achieved in a three-step paradigm. Following their isolation, stem/progenitor cells are expanded in neurosphere culture. Single cells obtained from these neurospheres are sorted for the expression of surface markers by flow cytometry. Finally, putative stem cells from cell sorting will be subjected to the so-called neural colony-forming cell assay, which allows discrimination between stem and progenitor cells. At the end of this chapter we will also describe how to identify neural stem cells in vivo. PMID- 21618084 TI - Magnetic resonance imaging of stem cell migration. AB - Noninvasive cellular imaging allows the real-time tracking of grafted cells as well as the monitoring of their migration. In this review, we will focus on cell tracking using MRI, since MRI is noninvasive, clinically transferable, and displays good resolution, ranging from 50 MUm in animal experiments up to 300 MUm using whole body clinical scanners. In addition to information about grafted cells, MRI provides information about the surrounding tissue (i.e., lesion size, edema, inflammation), which may negatively affect graft survival or the functional recovery of the tissue. Transplanted cells are labeled with MR contrast agents in vitro prior to transplantation in order to visualize them in the host tissue. The chapter will focus on the use of superparamagnetic iron oxide nanoparticles (SPIO), because they have strong effects on T2 relaxation yet do not affect cell viability, and will provide an overview of different modifications of SPIO and their use in MR tracking in living organisms. PMID- 21618085 TI - Imaging of Schwann cells in vivo. AB - The ability to examine cells tagged with fluorescent proteins in vivo has led to exciting advances in molecular neurobiology. The integral role of Schwann cells in nerve regeneration is well characterized, but not until recently has dynamic imaging of these critical cells been possible. Unlike many static techniques, in vivo imaging of Schwann cells tagged with green fluorescent protein (GFP) allows for serial imaging and may provide important insights into their role in peripheral nerve regeneration. In this chapter, we describe a protocol for in vivo imaging of SC migration into nonfluorescent nerve grafts using transgenic S100-GFP mice. PMID- 21618086 TI - Imaging of embryonic stem cell migration in vivo. AB - Conventional reporter gene technology and histological methods cannot routinely be used to track the in vivo behavior of embryonic stem (ES) cells longitudinally after cellular transplantation. Here we describe a protocol for monitoring the in vivo survival, proliferation, and migration of ES cells without necessitating animal sacrifice. Stable ES cell lines containing double fusion (DF; enhanced green fluorescent protein and firefly luciferase) or triple fusion (TF; monomeric red fluorescent protein, firefly luciferase, and herpes simplex virus thymidine kinase) reporter genes can be established within 4-6 weeks by lentiviral transduction followed by fluorescence-activated cell sorting. The cell fate and behavior of these DF or TF ES cells can subsequently be tracked noninvasively by bioluminescence and microPET imaging for a prolonged period of time. PMID- 21618087 TI - In vivo germ line stem cell migration: a mouse model. AB - A stem cell niche is a specialized tissue environment that controls the proliferation and differentiation of its resident stem cells. The functions of these structures have been well characterized in adult organisms. In particular, the bone marrow stem cell niche in mammals has been amenable to analysis because of the ability of transplanted hematopoietic cells to home and to recolonize the bone marrow of an irradiated host. Despite clues from adult models, it remains unclear how stem cells become partitioned into appropriate niches during embryonic development. To examine the earliest steps in niche formation, we created an organ culture system to observe the development of primordial germ cells (PGCs), a migratory stem cell population that will eventually give rise to the gametes. Using this assay, we can watch PGCs as they migrate to colonize the developing gonads and can introduce growth factor agonists or antagonists to test the function of proteins that regulate this process. This provides an unprecedented opportunity to identify the cellular and molecular interactions required for the formation of the germ cell niche. PMID- 21618088 TI - Live microscopy of neural stem cell migration in brain slices. AB - In the developing central nervous system (CNS), neural stem cells undergo a complex series of -morphogenetic and motile events. Errors in neural stem cell proliferation or migration cause serious brain developmental disorders. However, the relative importance of each step in neurogenesis and migration and the identity of genes affecting these processes has only begun to be explored. Using live imaging in brain slices, neural stem cells and their progeny labeled by in utero gene transfer can be monitored at high spatial and temporal resolution for as long as several days. Cell cycle progression, mitosis, morphogenesis, and migratory behavior can each be documented in detail. Furthermore, the behavior of subcellular structures, including nuclei, centrosomes, and microtubules, can also be observed using fluorescent marker proteins. This chapter describes the application of these approaches in combination with RNA interference to investigate normal developing brain and the role of genes involved in brain developmental disorders, such as lissencephaly. PMID- 21618089 TI - Whole embryo imaging of hematopoietic cell emergence and migration. AB - The use of transgenic mice in which tissue or lineage-specific, cell-restricted promoters drive fluorescent reporters has recently been reported as a means to follow the in vivo migration of various hematopoietic cells during murine development. At present there is limited ability of these approaches to image the emergence of the first hematopoietic cell subsets due to lack of unique markers that define those hematopoietic cells. We have utilized whole embryo analysis via immunostaining and confocal laser-scanning microscopic (CLSM) imaging to define the emergence of the first hematopoietic elements in the yolk sac of the developing conceptus. The methods employed to examine yolk sac hematopoiesis may be applied to hematopoietic cell emergence in the embryo proper or fetal liver in the generation of a complete map of hematopoietic ontogeny. PMID- 21618090 TI - Stem cell migration: a zebrafish model. AB - Compared with other vertebrate animal models, zebrafish (Danio rerio) has its superior advantages for studying stem cell migration. Zebrafish have similar tissues and organs as mammals, where tissue-specific stem cells reside in. Zebrafish eggs are externally fertilized and remain transparent until most of the organs are fully developed. This allows imaging stem cells in vivo very easily. Recently, a zebrafish double pigmentation mutant, casper, became a new popular imaging model in the zebrafish field due to its completely transparent bodies in adulthood. It has been used as an excellent model to study adult hematopoietic stem cell (HSC) in the transplantation setting. The unparalleled imaging power of zebrafish provides great opportunities of tracing stem cells in vivo in the developmental and regenerative context. In this chapter, we use HSC as an example and combine the powerful imaging techniques in zebrafish, to provide protocols for in vivo imaging fluorescence-labeled stem cell migration, stem cell fate tracing in zebrafish embryos, HSC transplantation, and in vivo imaging in both zebrafish embryos and adults. These techniques can also be applied to other types of stem cells in zebrafish embryos and adults. PMID- 21618091 TI - Imaging pluripotent cell migration in Drosophila. AB - Drosophila melanogaster offers a powerful system for the analysis of cell migration. In the embryo, -pluripotent cells of the mesodermal and endodermal primordia undergo epithelial-mesenchymal transitions and cell migration, while primordial germ cells migrate through an endodermal barrier to form the gonads. Visualisation of these migrations has traditionally been achieved by staining fixed embryos at different developmental stages or through live imaging of cells using tissue-specific expression of marker fluorescent proteins. More recently, photoactivatable fluorescence proteins have allowed the labelling of small groups of cells or single cells so that their migratory patterns and fate can be followed. By fusing the photoactivatable fluorescent protein to proteins that mark different subcellular components, it is now possible to visualise different aspects of the cells as they migrate. Here, we review previous studies of the migration of pluripotent embryonic cells and describe, in detail, methods for visualising these cells. PMID- 21618092 TI - Adhesion, migration, and homing of murine hematopoietic stem cells and progenitors. AB - Recent advances in cell biology have demonstrated the role of multiple signaling proteins in the transduction of external signals to cytoplasmic and nuclear effectors, controlling the movement and/or retention of hematopoietic stem cells and progenitors (HSC/P) within the bone marrow, with important clinical implications. Multiple assays have become routine in the analysis of adhesion to the microenvironment, migration toward chemoattractant gradients, and homing of HSC/P in the bone marrow in vivo. In this chapter, we analyze some of the most frequently used assays in our laboratory to explore the ability of HSC to migrate, adhere, and home in in vitro and in vivo assays. PMID- 21618093 TI - Methods to analyze the homing efficiency and spatial distribution of hematopoietic stem and progenitor cells and their relationship to the bone marrow endosteum and vascular endothelium. AB - The tracking of immunofluorescent labeled hematopoietic stem and progenitor cells (HSC/HPC) within the bone marrow (BM) cavity allows the assessment of the regulatory processes involved in transendothelial migration, trans-marrow migration, and finally lodgement into the HSC niche. This is of interest as the extracellular and cellular components involved in the regulation of HSC quiescence and differentiation are still not completely understood. Homing of transplanted HSC is the first critical step in the interaction between HSC and the microenvironment of the BM. As a consequence, murine models allowing the evaluation of the structural relationship between migrating HSC, the endosteal bone surface, and the vascular components of the BM enhance our understanding of hematopoietic regulation. PMID- 21618094 TI - Imaging hematopoietic stem cells in the marrow of long bones in vivo. AB - Hematopoietic stem and progenitor cells (HSPCs) are located in the bone marrow in zones of residence specialized in supporting them which are referred to as niches. It is in such a specialized niche that normal HSPCs are maintained to perform their self-renewal and differentiation duties in a highly controlled manner. One challenge in dissecting the functional significance of the complex cellular and molecular interactions in the niche is to link the types and qualities of cell-cell contacts to the intracellular signaling components involved in cell regulation. Attempts to study the interactions of HSPC with their niche eventually have to be performed in their natural location in vivo, as isolation of the cells from bone -marrow will disrupt the HSPC-niche interactions and thus not reveal functionally critical cell-cell -contacts. Intravital imaging of individual cells in the bone marrow has just recently been introduced, almost exclusively focusing on imaging inside the marrow of the calvaria. However, calvarial marrow is functionally distinct from marrow of long bones, the major source of HSPC for both physiology and study. To overcome these limitations, we developed a novel method for multiphoton intravital imaging of HSPC in the marrow of long bones. PMID- 21618095 TI - Studies of adult neural stem cell migration. AB - Adult neural progenitors originating is the subventricular zone (SVZ) proceed toward the olfactory bulb (OB) along a peculiar path of migration known as the rostral migratory stream. Once reaching the inner core of the bulb, neuroblasts migrate radially to various layers of the adult OB where they differentiate into local interneurons. We recently demonstrated that this latter form of migration is guided by blood vessels that serve as a scaffold for neural progenitor migration to their respective OB layers. Migration of progenitors along blood vessels can be visualized by time-lapse confocal imaging, which requires labeling of both migrating cells and blood vessels. In this article, we describe procedures to label neural progenitors in adult through in vivo stereotaxic cell tracer injection into the SVZ, and to visualize blood vessels through luminal filling with stabilized fluorescent dyes. Moreover, we illustrate the methods for acute brain slices preparation for imaging and procedures used during confocal time-lapse imaging to follow the movement of adult neural progenitors along blood vessels. PMID- 21618096 TI - Dissecting mesenchymal stem cell movement: migration assays for tracing and deducing cell migration. AB - Targeted migration is a necessary attribute for any gene delivery vehicle. Mesenchymal stem cells (MSC) have been used as effective delivery vehicles for treatments against cancer, graft versus host disease, -arthritis, multiple sclerosis, and many other diseases. MSC migrate toward sites of inflammation, however, the true migratory mechanism has yet to be elucidated. There are several receptors and respective chemokines known to be involved in the migration of the MSC. Further insight to MSC migration will be revealed both in vivo and in vitro through the application of migration assays from the most simple, to the more technologically demanding. PMID- 21618097 TI - Epithelial stem cells. AB - It is likely that adult epithelial stem cells will be useful in the treatment of diseases, such as ectodermal dysplasias, monilethrix, Netherton syndrome, Menkes disease, hereditary epidermolysis bullosa, and alopecias. Additionally, other skin problems such as burn wounds, chronic wounds, and ulcers will benefit from stem cell-related therapies. However, there are many questions that need to be answered before this goal can be realized. The most important of these questions is what regulates the adhesion of stem cells to the niche versus migration to the site of injury. We have started to identify the mechanisms involved in this decision-making process. PMID- 21618099 TI - The role of receptor tyrosine kinases in primordial germ cell migration. AB - During embryonic development in Drosophila, rodents, and other organisms, primordial germ cells (PGCs) migrate from their points of origin to the nascent gonads, where they give rise to germ line stem cells. Receptor tyrosine kinase (RTK) activity is required for normal migration of primordial germ cells in both Drosophila and rodents. In this chapter, we discuss in vivo as well as in vitro methods which have been used to elucidate the role of the RTK Torso in Drosophila germ cell migration. Included are protocols for embryo collection, fixation, and immunostaining; the dominant female sterile technique; in vitro culture and observation of PGCs; pole cell transplantation; and labeling of pole cells for in vivo observation. PMID- 21618098 TI - Pathways implicated in stem cell migration: the SDF-1/CXCR4 axis. AB - The hallmark of hematopoietic stem and progenitor cells (HSPCs) is their motility, which is essential for their function, such as recruitment upon demand. Stromal Derived Factor-1 (SDF-1, CXCL12) and its major receptor CXCR4 play major roles in stem cell motility and development. In vitro migration assays, implicating either gradients or cell surface-bound forms of SDF-1, are easy to perform and provide vital information regarding directional and random stem cell motility, which correlate with their repopulation potential in clinical and experimental transplantations. In vivo stem cell homing to the bone marrow, their retention, engraftment, and egress to the circulation, all involve SDF-1/CXCR4 interactions. Finally, other stem cell features such as stem cell survival and proliferation, are also dependent on the SDF-1/CXCR4 axis. PMID- 21618100 TI - Rho GTPases in hematopoietic stem/progenitor cell migration. AB - Rho GTPases including RhoA, Rac1, and Cdc42 are a class of intracellular signaling proteins critical for the regulation of cytoskeleton organization, adhesion, and migration. Molecular mechanisms of mammalian cell migration were first revealed in fibroblasts where RhoA, Rac1, and Cdc42 facilitate in the multistep process including establishment and maintenance of polarity, formation of actin-rich protrusions, remodeling of adhesive contacts, and generation of force. In hematopoietic stem/progenitor cells, Rho GTPases relay signals from chemokines and cytokines such as SDF-1alpha and SCF to the actin and microtubule cytoskeleton through effector kinases and/or adaptor molecules that affect adhesion or transcription. Comprehensive use of murine conditional gene knockout technology combined with biochemical approaches in recent studies allows for physiologically relevant investigations of the involvement of Rho GTPases in hematopoietic stem/progenitor cell migration, providing important mechanisms for the stem/progenitor maintenance. PMID- 21618101 TI - Serendipity in scientific discoveries: some examples in glycosciences. PMID- 21618102 TI - My 30-year devotion to the N-linked oligosaccharide structures. PMID- 21618103 TI - Human blood group ABH/Ii, Le(a,b,x,y), and sialyl Le (a,x) glycotopes; internal structures; and immunochemical roles of human ovarian cyst glycoproteins. PMID- 21618104 TI - Lewis glyco-epitopes: structure, biosynthesis, and functions. PMID- 21618105 TI - A unique endo-beta-galactosidase that cleaves both blood group A and B glycotopes. PMID- 21618106 TI - Recognition roles of mammalian structural units and polyvalency in lectin--glycan interactions. PMID- 21618107 TI - Adhesion/growth-regulatory galectins: insights into their ligand selectivity using natural glycoproteins and glycotopes. PMID- 21618108 TI - Glycotope structures and intramolecular affinity factors of plant lectins for Tn/T antigens. PMID- 21618109 TI - The five bacterial lectins (PA-IL, PA-IIL, RSL, RS-IIL, and CV-IIL): interactions with diverse animal cells and glycoproteins. PMID- 21618110 TI - On the differential sialic acid specificity of lectins from different parts of Saraca indica. PMID- 21618111 TI - Regulation of lectin production by the human pathogens Pseudomonas aeruginosa and Chromobacterium violaceum: effects of choline, trehalose, and ethanol. PMID- 21618112 TI - Non-carbohydrate-mediated interaction of lectins with plant proteins. PMID- 21618113 TI - Novel concepts about the role of lectins in the plant cell. PMID- 21618114 TI - Role of gangliosides and plasma membrane-associated sialidase in the process of cell membrane organization. PMID- 21618115 TI - 9-O-Acetyl GD3 in lymphoid and erythroid cells. PMID- 21618116 TI - GM3 Upregulation of matrix metalloproteinase-9 possibly through PI3K, AKT, RICTOR, RHOGDI-2, and TNF-A pathways in mouse melanoma B16 cells. PMID- 21618117 TI - Pathological roles of ganglioside mimicry in Guillain-Barre syndrome and related neuropathies. PMID- 21618118 TI - Structure and function of glycolipids in thermophilic bacteria. PMID- 21618119 TI - Lipooligosaccharides of Neisseria species: similarity between N. polysaccharea and N. meningitidis LOSs. PMID- 21618120 TI - Roles for N- and O-glycans in early mouse development. PMID- 21618121 TI - Glycobiology in the field of gerontology (glycogerontology). PMID- 21618122 TI - Galectins in regulation of apoptosis. PMID- 21618123 TI - Avian and human influenza virus receptors and their distribution. PMID- 21618124 TI - Importance of a factor VIIIc-like glycoprotein expressed in capillary endothelial cells (eFactor VIIIc) in angiogenesis. PMID- 21618125 TI - Mucin O-glycan branching enzymes: structure, function, and gene regulation. PMID- 21618126 TI - Studying carbohydrate self-recognition in marine sponges using synthetic aggregation factor epitopes. PMID- 21618127 TI - Docking of chitin oligomers and Nod factors on lectin domains of the LysM-RLK receptors in the Medicago-Rhizobium symbiosis. PMID- 21618128 TI - O-Acetylated sialic acids and their role in immune defense. PMID- 21618129 TI - Sialylated and sulfated carbohydrate ligands for selectins and siglecs: involvement in traffic and homing of human memory T and B lymphocytes. PMID- 21618130 TI - Diversity of natural anti-alpha-galactosyl antibodies in human serum. PMID- 21618131 TI - Significance of serum glycoprotein profiles in spontaneous tolerance after liver allograft transplantation. PMID- 21618132 TI - Hematogenous metastasis: roles of CD44v and alternative sialofucosylated selectin ligands. PMID- 21618133 TI - Regulation of glycosyltransferase genes in apoptotic breast cancer cells induced by L: -PPMP and cisplatin. PMID- 21618134 TI - Aberrant glycosphingolipid expression and membrane organization in tumor cells: consequences on tumor-host interactions. PMID- 21618135 TI - Human KDN (deaminated neuraminic acid) and its elevated expression in cancer cells: mechanism and significance. PMID- 21618136 TI - Polysialic acid bioengineering of cancer and neuronal cells by N-acyl sialic acid precursor treatment. PMID- 21618137 TI - Synthesis of hemagglutinin-binding trisaccharides. PMID- 21618138 TI - Fabrication and applications of glyconanomaterials. PMID- 21618139 TI - Glycan arrays to decipher the specificity of plant lectins. PMID- 21618140 TI - Targeting C-type lectin for the treatment of flavivirus infections. PMID- 21618141 TI - Effect of growth hormone on steroid content, proliferation and apoptosis in the chicken ovary during sexual maturation. AB - The present study was undertaken to examine in vivo the effect of growth hormone (GH) on progesterone and estradiol levels and on cell proliferation and apoptosis in the chicken ovary during sexual maturation. Hy-Line chickens (10 weeks old) were injected three times a week with 200 MUg recombinant chicken GH (cGH) per kilogram body weight until sexual maturity. GH treatment significantly increased ovarian weight at 16 weeks of age, i.e., ~1 week before onset of egg laying. The progesterone content in the ovary just before and at the time of sexual maturity and the estradiol content before onset of egg laying were also elevated after cGH injections. The highest number of proliferating (positive for proliferating cell nuclear antigen) and apoptotic (positive for terminal-deoxynucleotidyl transferase-mediated dUTP nick-end labeling) cells was found in the ovarian stroma and white follicles (>1-4 mm diameter), whereas the lowest number of these cells was detected in yellow (>8-30 mm) follicles. Administration of cGH significantly stimulated cell proliferation and inhibited cell apoptosis in the ovarian stroma and small ovarian follicles. The number of ovarian follicles and the weight of the ovary prior to the first oviposition were also higher in cGH injected hens. Thus, prior to and after the onset of egg laying, GH participates in the growth, maturation and hormonal activity of ovarian follicles in the chicken, via the regulation of steroidogenesis, proliferation and apoptosis processes. PMID- 21618142 TI - TGF-beta signal transduction spreading to a wider field: a broad variety of mechanisms for context-dependent effects of TGF-beta. AB - Transforming growth factor (TGF)-beta signaling is involved in almost all major cell behaviors under physiological and pathological conditions, and its regulatory system has therefore been vigorously investigated. The fundamental elements in TGF-beta signaling are TGF-beta ligands, their receptors, and intracellular Smad effectors. The TGF-beta ligand induces the receptors directly to phosphorylate and activate Smad proteins, which then form transcriptional complexes to control target genes. One of the classical questions in the field of research on TGF-beta signaling is how this cytokine induces multiple cell responses depending on cell type and cellular context. Possible answers to this question include cross-interaction with other signaling pathways, different repertoires of Smad-binding transcription factors, and genetic alterations, especially in cancer cells. In addition to these genetic paradigms, recent work has extended TGF-beta research into new fields, including epigenetic regulation and non-coding RNAs. In this review, we first describe the basic machinery of TGF beta signaling and discuss several factors that comprise TGF-beta signaling networks. We then address mechanisms by which TGF-beta induces several responses in a cell-context-dependent fashion. In addition to classical frames, the interaction of TGF-beta signaling with epigenetics and microRNA is discussed. PMID- 21618143 TI - Indigestible dextrin stimulates glucoamylase production in submerged culture of Aspergillus kawachii. AB - Submerged batch cultures of Aspergillus kawachii grown on indigestible dextrin were investigated for potential improvements in glucoamylase (GA) production. In flask culture, specific GA productivities per dry weight biomass using dextrin and indigestible dextrin were 11.0 and 56.1 mU/mg-DW, respectively. Indigestible dextrin was a poor substrate for enzymatic hydrolysis. Rates of glucose formation from dextrin and indigestible dextrin by enzymatic hydrolysis were 0.477 and 0.100 mg-glucose/ml/h, respectively. For this reason, residual glucose concentrations in batch cultures grown on indigestible dextrin remained below 1.32 mg/ml where glucose-limiting conditions were easily maintained. Batch culture using indigestible dextrin had the same residual glucose profile as dextrin fed-batch culture, and nearly the same GA activity was obtained after 42.5 h of growth. However, between 42.5 and 66 h, the GA production rate of the indigestible dextrin batch culture (11.5 mU/ml/h) was higher than that of the dextrin fed-batch culture (6.5 mU/ml/h). During this period, a high amount of residual maltooligosaccharide was detected in the culture supernatant grown on indigestible dextrin. The high GA productivity observed in the indigestible dextrin batch culture may have resulted from the absence of glucose and the simultaneous presence of maltooligosaccharides throughout growth. PMID- 21618144 TI - An interesting electrocardiogram. PMID- 21618145 TI - 50 Years ago in CORR: Arthrography of the shoulder Joint Robert L. Samilson MD, Robert L. Raphael MD, Lawrence Post MD, Charles Noonan MD, Evelyn Siris MD, and Frank L. Raney Jr, MD CORR 1961;20:21-32. PMID- 21618146 TI - Adiposity, joint and systemic inflammation: the additional risk of having a metabolic syndrome in rheumatoid arthritis. AB - Adiposity is a predisposing condition to atherosclerosis, and rheumatoid arthritis (RA) also predisposes to accelerated atherosclerosis. Adiposity is one of the key features of the metabolic syndrome (MetS) and it is well recognised that a metabolic syndrome (and fat tissue) is a major player in this complex network. Endothelial dysfunction and carotid intima-media thickness, early pre clinical markers of atherosclerosis which are the main determinants of cardiovascular (CV) morbidity and mortality, occur early on in RA. RA patients have an incidence of CV diseases at least two times higher than the general population. MetS and RA have a low and a severe-moderate degree of inflammation in common, respectively. Adipose tissue has emerged as a dynamic organ that releases several inflammatory and immune mediators (adipokines). In addition, fat has been recognised as a producer of B cell activating factor (BAFF) and of chemerin, an inducer at the chondrocyte level of IL1beta, TNFalpha, IL6, IL8 and MMP13, thus possibly contributing to cartilage damage. Since fat produces inflammation, to obtain a full control of the CV risk in RA, data suggest that it is therefore mandatory to have a "tight control" of both RA and MetS-related inflammation, especially if RA presents MetS as a co-morbidity. PMID- 21618147 TI - Incidence, aetiology and pattern of mandibular fractures in central Switzerland. AB - PURPOSE: The two major causative factors for mandibular fractures, as stated in the literature, are either interpersonal violence or motor vehicle accidents. The purpose of this study was to describe epidemiological trends of mandibular fractures in Switzerland. A special emphasis was directed towards the potential impact of socio-economic standards on the mechanism and pattern of mandible fractures. PATIENTS AND METHODS: A database of patients aged over 16 years who had been diagnosed with a mandibular fracture between January 2000 and December 2007 at the University Hospital of Bern, Switzerland's largest Cranio Maxillofacial-Surgery Centre, was retrospectively reviewed. Patients' data including gender, age, mechanism of accident, fracture site and associated injuries were analysed and compared with previously published data. RESULTS: There were a total of 420 patients with 707 mandibular fractures. The two most common causes of injury were road traffic accidents (28%) and various types of sports injuries (21%). A total of 13% of the patients were under the influence of alcohol or drugs at admission. Fractures were predominantly situated in the condyle/subcondyle (43%) and in the symphysis/parasymphysis region (35%). Occurrences of fractures in the angle and in the body were low, at 12% and 7% respectively. CONCLUSION: In contrast to other highly developed countries, sports and leisure-related accidents outnumbered motor vehicle accidents and altercations. The data presented here supports the assumption of a correlation of trauma cause and fracture pattern. PMID- 21618148 TI - Blinded by the light. PMID- 21618149 TI - Algorithm for detection of small-bowel metastasis in malignant melanoma of the skin. AB - BACKGROUND AND STUDY AIM: The aim of this study was to develop an algorithm to detect small-bowel metastasis (SBM) of melanoma by sequential laboratory parameters and pan-intestinal endoscopy (PIE) including video capsule endoscopy (VCE). PATIENTS AND METHODS: A total of 390 melanoma patients (AJCC stage I/II/III/IV, 140/80/121/49) were screened for signs of intestinal blood loss (fecal occult blood test [FOBT] or overt bleeding) in an open, multicenter, prospective study, and those who were positive underwent PIE. Independent of the presence of intestinal bleeding, all stage IV patients were offered PIE. Follow up was obtained in 357 patients (91.5 %) for a median of 16 months. We undertook to identify possible associations between SBM and clinical and laboratory data. Survival data were analyzed with regard to clinical and laboratory data and small bowel findings. RESULTS: Intestinal blood loss was suspected in 49 of 390 patients (12.6 %), 38 of whom (77.6 %) agreed to undergo endoscopy. In 10 patients, SBM was detected by VCE (intention-to-diagnose, 20.4 %; AJCC III, n = 2; AJCC IV, n = 8). The SBM was resected in five patients. Total detection rates of SBM were 14 of 49 patients in stage IV (28.6 %, intention-to-diagnose), 2 of 121 in stage III (1.7 %), and 0 in stage I/II. In FOBT-positive patients, SBM detection rates were 72.7 %, 14.3 %, and 0 % in tumor stages IV, III, and I/II, respectively. Positive FOBT proved to be an independent negative prognostic factor for total survival in stage III and IV melanoma. CONCLUSIONS: SBMs are frequent in advanced melanoma. In stage III patients, screening for intestinal blood loss by PIE may help to identify SBMs. In stage IV, indication for PIE should depend on the individual consequences of detecting SBM, but not on bleeding symptoms alone. PMID- 21618150 TI - Factors that predict bleeding following endoscopic mucosal resection of large colonic lesions. AB - BACKGROUND AND STUDY AIMS: Endoscopic mucosal resection (EMR) for large colonic laterally spreading tumors (LSTs) is a safe, efficacious, and cost-effective treatment. The most common serious complication is delayed bleeding, which reduces these advantages, but consensus guidelines for large-polyp EMR do not exist. PATIENTS AND METHODS: Data from two large prospective intention-to-treat studies of EMR for colonic LSTs 20 mm or greater in size were analyzed. Data collection was comprehensive, and included patient and lesion characteristics. EMR technique and cessation of anticoagulant and antiplatelet therapy was standardized. Clinically significant delayed bleeding was defined as that requiring hospital admission. RESULTS: EMR was performed on 302 lesions in 288 patients. There was clinically significant delayed bleeding in 21 cases (7 %). Ten underwent colonoscopy. One required angiography. One required surgery after perforation following hemostatic clip placement. There were no deaths. Risk factors for bleeding on multivariate analysis were right colon location [adjusted odds ratio (OR) 4.4, P = 0.01], use of aspirin (OR 6.3, P = 0.005), and age (OR per decade of age 1.70). All bleeds occurred before aspirin was restarted. Patient characteristics, including ASA grade and co-morbidity type, were not predictive. Despite requiring more complex EMR, larger lesion size ( P = 0.2), multiple excisions rather than en bloc resection ( P = 0.1), polyp morphology ( P = 0.2), and previous attempts ( P = 0.5), were not associated with increased risk. CONCLUSIONS: Proximal lesion location is a highly significant risk for clinically significant delayed bleeding following colonic EMR, and this knowledge could form the basis of a targeted therapeutic trial. Recent aspirin use also increases bleeding risk--specific consensus guidelines in this area are required for colonic EMR. PMID- 21618151 TI - Elevated stricture rate following the use of fully covered self-expandable metal biliary stents for biliary leaks following liver transplantation. AB - BACKGROUND: Biliary leaks and strictures are common complications after liver transplantation and can be managed surgically or endoscopically. Endoscopic management using fully covered self-expandable metal stents (FCSEMS) might provide some advantages over the commonly used plastic stents in the management of bile leaks after liver transplantation. METHODS: Between December 2006 and January 2009, 17 liver transplant recipients underwent placement of a FCSEMS for treatment of biliary leaks. RESULTS: FCSEMS were deployed at median of 18 days (range: 6 - 160) after liver transplantation and left in place for a median of 102 days (range: 35 - 427), with a median follow-up after FCSEMS removal of 407 days (range: 27 - 972). Long-term leak control was obtained in all but one patient. Complications included 6 clinically significant biliary strictures (35 %), which were treated with repeat stent placement, and two clinically insignificant strictures (12 %) which required no intervention. Additionally, three patients (18 %) had biliary ulcerations after stent removal, confirmed by choledochoscopy, and were managed conservatively. Two patients required repeat liver transplantation due to hepatic artery thrombosis, and one patient died from sepsis unrelated to FCSEMS stenting. CONCLUSIONS: FCSEMS treat biliary leaks effectively, but carry a relatively high stricture risk in patients who have received liver transplants. FCSEMS cannot be recommended for management of biliary leaks following liver transplantation at this point. PMID- 21618152 TI - Asian minimal-change esophagitis in the era of high-definition endoscopy. PMID- 21618155 TI - Concurrent training and pulmonary function in smokers. AB - This study compared the effects of aerobic, resistance and concurrent aerobic and resistance training on pulmonary function and cardiorespiratory endurance in at risk smokers. 50 sedentary, male smokers with pulmonary function impairments at risk for developing chronic lung diseases were randomly assigned to an aerobic (AerG;n=12), resistance (ResG;n=13), concurrent (ConG;n=13) or non-exercising control (NexG;n=12) group for 16 weeks. AerG subjects performed 45 min of aerobic exercise at 60%HR(max), ResG subjects performed 8 resistance exercises at 60%1-RM for 3 sets, 15 repetitions while ConG subjects performed both aerobic and resistance exercises. ANOVA revealed no significant difference between the groups in their pre-/post-test changes for FEV(1)/FVC ratio (AerG:-4.13%; ResG:-2.13%; ConG:-0.56%); FEF-50 (AerG:-4.59%; ResG:-7.62%; ConG:5.76%), FEF-75 (AerG:-2.36%; ResG:-7.62%; ConG:10.71%) and FEF 25-75 (AerG:-3.53%; ResG:-6.43%; ConG:7.63%). Significant differences were found between the groups in their pre-/post-test changes for FVC (AerG:8.05%; ResG:7.22%; ConG:11.55%), FEV(1) (Aer:9.60%;ResG:5.13%; ConG:12.10%), PEF (AerG: 11.29%; ResG:7.49%; ConG:20.18%), PIF (AerG:24.80%; ResG:19.41%; ConG:28.15%), IVC (AerG: 9.04%; ResG: 6.21%; ConG:16.35%), FEF-25 (AerG:5.88%; ResG:5.37%; ConG:11.88%) and cardiorespiratory fitness (AerG:25.44%; ResG:11.59%; ConG:22.83%). Post-hoc analysis revealed concurrent and aerobic training were equally effective at improving PIF and cardiorespiratory fitness with concurrent training most effective at improving FVC, FEV(1), PEF, IVC and FEF-25. This suggests synergy between aerobic and resistance exercise in preventing or reducing the detrimental effects of smoking while gaining the unique benefits of each mode of exercise. PMID- 21618156 TI - Does upper body strength and power influence upper body Wingate performance in men and women? AB - The aim of this study was to determine the influence of muscular strength and power on upper body Wingate performance in men and women. Muscular strength (1 repetition maximum bench press), muscular power (bench throws) and upper body anaerobic performance (Wingate Anaerobic Test (WAnT)) was assessed in 24 men and 16 women. Men had significantly ( P<0.001) higher absolute and relative peak and mean power and blood lactate concentration during the WAnT compared to their female counterparts. Men also produced significantly ( P<0.001) higher strength and absolute and relative peak and mean power during the bench press and throw, respectively, compared to the female participants. For men body mass and mean power produced during the bench throw explained approximately 84% and 87% of the variance in Wingate peak ( P<0.001 and P=0.039, respectively) and mean ( P<0.001 and P=0.028, respectively) power. For women mean power produced during the bench throw explained approximately 72% and 52% of the variance in Wingate peak ( P=0.002) and mean ( P=0.017) power, respectively. For men body mass and to a lesser extent muscular power best predicts upper body Wingate performance while for women only muscular power predicts upper body Wingate performance. PMID- 21618157 TI - Fluctuating asymmetry as a predictor for rowing ergometer performance. AB - Fluctuating asymmetry (FA) can be defined as an organism's deviation from perfect bilateral symmetry. FA has been of interest to evolutionary biologists as it may be indicative of the ability of an individual to express its genotype in a stable manner. Asymmetry has been shown to correlate with success in both intra- and inter-sexual selection in various species, including humans. A growing body of knowledge is emerging concerning the relationship between asymmetry and sporting ability. This study seeks to expand upon understanding of developmental stability and athletics by investigating the association between asymmetry and performance in rowers. Both male and female competitors from a range of abilities were tested (76 males and 70 females), with asymmetry being determined through bilateral digit measurements and performance through personal best time over 2000 m on the Concept 2 indoor rowing ergometer. Significant negative correlations were observed between asymmetry and 2000 m ergometer performance in both males and females. The relationship remained significant after adjustment for rowing experience and height. These results suggest that asymmetry may serve as a predictor of potential ability in the sport of rowing. The cause for the association between asymmetry and sporting performance has yet to be identified. PMID- 21618158 TI - Influence of cycle ergometer type and sex on assessment of 30-second anaerobic capacity and power. AB - This study examined the influence of cycle ergometer type and sex on assessment of 30-s anaerobic capacity and power. 41 healthy adults performed a 30-s anaerobic cycle test using a mechanically- (ME) and air-braked (AE) ergometer in a randomised order, approximately 7 days apart. Peak heart rate (HR) and rating of perceived exertion were similar between tests with peak HR greater for females compared to males (187.0 +/- 9.1 vs. 180.8 +/- 9.9 bpm, p<0.05). Peak power (1 100 +/- 330 vs. 802 +/- 225 W), mean power (793 +/- 223 vs. 587 +/- 156 W) and total work (23.8 +/- 6.7 vs. 17.6 +/- 4.7 kJ) were greater for AE compared to ME (p<0.001) and greater for males compared to females (p<0.001). The mean difference for anaerobic capacity and power between AE and ME were similar for males and females (37-41% vs. 33-35%, p>0.05). Peak lactate was greater for AE compared to ME (16.1 +/- 3.4 vs. 14.8 +/- 2.9 mmol.L (-1); p<0.05) and greater for males compared to females (16.2 +/- 3.5 vs. 14.6 +/- 2.7 mmol.L (-1); p<0.05). The current study demonstrated that anaerobic power and capacity were substantially greater when assessed using AE compared to the traditional ME with the difference between ergometer types unaffected by sex. Ergometer type should be considered when comparing anaerobic results across populations and/or studies. PMID- 21618159 TI - Adapting workload improves the measurement of heart rate recovery. AB - Heart rate after a standardized test varies with a change in training status, possibly compromising the accuracy of measuring changes in heart rate recovery (HRR). The aim of this study was to determine if a change in the exercise intensity would result in a change in heart rate recovery and/or the accuracy of the heart rate recovery measurement. 31 subjects performed 4 submaximal running tests (HIMS). Based on the heart rate after the first HIMS, subjects either completed 4 identical HIMS (SAME (n=9)), 2 standard and 2 faster HIMS (FASTER (n=10)) or 2 standard and 2 slower HIMS (SLOWER (n=12)). Although no changes in heart rate recovery were found when the HIMS protocol was adapted, lower coefficients of variation (CV) and typical errors of measurement (TEM) were found in the SLOWER (CV: 11 +/- 7 to 5 +/- 3% ( P=0.025)), TEM: 6 to 3 beats and FASTER group (CV: 11 +/- 7 to 4 +/- 3% ( P=0.048), TEM: 7 to 3 beats). To ensure the highest level of sensitivity in detecting meaningful changes in HRR over time, submaximal testing protocols should target exercise intensities ranging in between 86-93% of heart rate maximum. PMID- 21618160 TI - Exercise improved rat metabolism by raising PPAR-alpha. AB - Based on the importance of exercise and crucial role of liver in metabolism, the aim of this study was to determine whether the expression of peroxisome proliferator-activated receptor (PPAR)-alpha, gamma and adiponectin receptor 2 in OLETF rat liver were altered in conjunction with improved metabolism with exercise training. OLETF rats were randomly assigned to 2 groups: sedentary control group (n=26), and long-term exercise-trained group (n=26). Full data were available on 32 OLETF rats (16 for each group). Adiponectin, glucose, insulin, triglyceride and cholesterol were assessed. Livers were taken to determine the expression of PPAR-alpha, gamma and adiponectin receptor 2. Compared with sedentary control group, fasting glucose (9.38 +/- 2.99 mmol/L vs. 7.32 +/- 1.76 mmol/L, P<0.05), triglyceride (1.73 +/- 0.34 mmol/L vs. 0.89 +/- 0.12 mmol/L, P<0.05) and cholesterol (4.41 +/- 0.75 mmol/L vs. 2.13 +/- 0.32 mmol/L, P<0.05) were substantially reduced after exercise, which significantly correlated with increased PPAR-alpha (P<0.05) in liver. The expression of PPAR-alpha upstream and target genes, including hepatocyte nuclear factor-4 (HNF4), carnitine palmitoyl transferase 1 (CPT-1), catalase (CAT) and ATPbinding cassette transporter A1 (ABCA1) also increased significantly. Therefore, our findings suggest that increased PPAR-alpha expression in OLETF rats liver is a contributory factor to the exercise-related improvements in whole-body metabolism. PMID- 21618161 TI - Changes in blood circulation of the contralateral Achilles tendon during and after acupuncture and heating. AB - The purpose of this study was to investigate the effects of acupuncture and heating (application of hot pack) treatments on blood circulation in the contralateral Achilles tendon. During the treatments (10 min for acupuncture, 20 min for heating) and recovery period (40 min), the blood volume (THb) and oxygen saturation (StO2) of the treated and the non-treated tendons were measured using red laser lights. During both treatments, THb and StO2 of the treated tendon increased significantly from the resting level. The increased THb and StO2 of the treated tendon were maintained until the end of the recovery period after removal of the acupuncture needle, although these values decreased after removal of the hot pack. Although THb of the non-treated sides did not change during both acupuncture and heating treatments, it increased gradually after removal of the acupuncture needle or the hot pack. For both treatments, the amount of increase in THb of the non-treated tendon was significantly correlated to that of the treated tendon during the last phase of recovery period. These results obtained from the healthy subjects imply that blood circulation in the injured tendon in a plaster cast may be improved by applying acupuncture or heating treatments to the contralateral healthy limb. PMID- 21618162 TI - Increased average longevity among the "Tour de France" cyclists. AB - It is widely held among the general population and even among health professionals that moderate exercise is a healthy practice but long term high intensity exercise is not. The specific amount of physical activity necessary for good health remains unclear. To date, longevity studies of elite athletes have been relatively sparse and the results are somewhat conflicting. The Tour de France is among the most gruelling sport events in the world, during which highly trained professional cyclists undertake high intensity exercise for a full 3 weeks. Consequently we set out to determine the longevity of the participants in the Tour de France, compared with that of the general population. We studied the longevity of 834 cyclists from France (n=465), Italy (n=196) and Belgium (n=173) who rode the Tour de France between the years 1930 and 1964. Dates of birth and death of the cyclists were obtained on December 31 (st) 2007. We calculated the percentage of survivors for each age and compared them with the values for the pooled general population of France, Italy and Belgium for the appropriate age cohorts. We found a very significant increase in average longevity (17%) of the cyclists when compared with the general population. The age at which 50% of the general population died was 73.5 vs. 81.5 years in Tour de France participants. Our major finding is that repeated very intense exercise prolongs life span in well trained practitioners. Our findings underpin the importance of exercising without the fear that becoming exhausted might be bad for one's health. PMID- 21618163 TI - [Archaeological and forensic ophthalmopathology]. PMID- 21618165 TI - [Acute abdomen in a 32-year-old woman]. PMID- 21618166 TI - Relevance and methods of interventional breast sonography in preoperative axillary lymph node staging. AB - Reduction of therapy-induced morbidity is an important goal for the improvement of the quality of breast cancer treatment. The introduction of sentinel lymph node biopsy (SLNB) significantly contributed to the reduction of surgery-induced morbidity in the shoulder-arm region. However, a clinically positive nodal status is still considered a contra-indication for SLNB. The current data constellation clearly shows that the clinical and also the sonographic malignancy assessment is insufficient for the accurate identification of the nodal status. A merely clinical and/or image-based appraisal of the axillary lymph node status would lead to overtreatment due to unnecessary axillary dissection in approximately 40 % of patients. In order to reduce the rate of unnecessary axillary dissection (AD), pretreatment interventional clarification is necessary to provide more detailed information about the histological condition of the lymph node. Comparing the currently available methods, fine needle aspiration (FNA) is the best in terms of cost and time requirement, practicability and complication rate. However, considering the sensitivity, it is inferior to ultrasound-guided core needle biopsy (CNB). Thus, a negative FNA outcome requires further clarification, which possibly can be performed with CNB. With a specificity of nearly 100 % and therefore a low rate of false positive cases for FNA, complete AD can be indicated by a positive FNA result. In the context of the interventional procedure, it must be stressed that FNA requires a high level of expertise on the part of both the examiner and the cytologist. The prerequisite for optimal interventional diagnostics of lymph nodes is an adequate sonographic assessment on the basis of standardized sonomorphological criteria. PMID- 21618167 TI - [Scrotal ultrasound in children and adolecents with duplex Doppler analysis of intratesticular arteries]. AB - PURPOSE: Scrotal ultrasound, color Doppler and spectral Doppler analysis of intratesticular arteries are the most common and important examinations in boys with scrotal pain. In the existing literature information concerning feasibility and reference values of color Doppler and spectral Doppler in small testes is inconsistent. MATERIALS AND METHODS: In the present study 102 boys from 2 days to 16 years old without present or anamnestic scrotal disease were examined in a standardized manner. Using linear scanners (9 - 14 mHz), testicular volumetry and spectral Doppler analysis of typical intratesticular arteries were performed and the paratesticular structures were examined. For analysis we grouped the testes by volume and compared the measured values V. max syst., V. max enddiast., and RI. RESULTS: In all test subjects a complete examination with spectral Doppler analysis of intratesticular arteries could be performed. With increasing testicular volume, there is a linear increase in blood flow velocities V. max syst. and V. enddiast. Irrespective of age and testicular volume, the RI of the intratesticular arteries is 0.54 +/- 0.08. CONCLUSION: In contrast to published data, this study shows that color Doppler and spectral Doppler of testicular arteries can be regularly performed even in small testes of less than 1 ml. Reference values for blood flow velocities and RI were found and should improve the diagnostic value of testicular ultrasound examinations in children. PMID- 21618168 TI - [Pulmonary Graft versus Host Disease]. PMID- 21618179 TI - [Temporary legal waiver in partner resignation from medical practice group]. PMID- 21618181 TI - [Editorial. Parasitic diseases in Turkey]. PMID- 21618182 TI - [Comparison of polymerase chain reaction with wet mount and culture methods for the diagnosis of trichomoniasis]. AB - OBJECTIVE: Wet mount, culture and staining methods are generally used in the diagnosis of trichomoniasis. Polymerase chain reaction (PCR) methods with different primer pairs have been tested and used for research in recent years. METHODS: In this study, T. vaginalis was tested for in the vaginal samples of 102 patients referred to obstetrics and gynecology polyclinic of Aydin Obstetrics and Children Hospital for various reasons, with direct microscopy, culture and PCR with primers targeting Tv-E650. In addition, PCR was applied to 20 T. vaginalis strains that were isolated from patients who were previously diagnosed with vaginitis. RESULTS: Of 102 samples, T. vaginalis was found to be positive in 2.94, 4.90 and 4.90% with wet mount, TYM medium and PCR respectively. The positivity rate reached 5.88% using the 3 methods together. All 20 strains isolated from patients with vaginitis were reported as positive by the PCR method. CONCLUSION: The wet mount had 60% sensitivity and 100% specificity, while PCR showed 80% sensitivity and 97.95% specificity when compared with the culture method, which is accepted as the "gold standard". The PCR method was performed for the first time as a diagnostic assay for trichomoniasis in this study and it is concluded that it can be used routinely for its diagnosis. PMID- 21618183 TI - The investigation of Entamoeba histolytica prevalence in some villages of Sivas by ELISA method. AB - OBJECTIVE: Humans may be infected with three morphologically identical Entamoeba species; pathogenic E. histolytica, commensal E. moshkowskii and E. dispar. The aim of the present study was to determine the true prevalence of the E. histolytica using native lugol, trichrome staining and a monoclonal antigen detection kit (ELISA kit E. histolytica-II; Techlab, Inc., Blacksburg, VA) among primary school children living in the rural areas around Sivas. METHODS: A total of 1449 stool samples were examined by native lugol and Trichrome staining, and 312 (22%) samples were positive for one or more parasite species. Additionally, 22 (1.5%) stool samples were found to be positive for the presence of E. histolytica/dispar cysts, and these samples were further examined by E. histolytica specific antigen based ELISA. RESULTS: As a result, ELISA test gave negative reactions for all the samples. Also, there was no cross reaction with other luminal protozoa such as E. coli, G. intestinalis, B. hominis and I. butschlii. CONCLUSION: The data reveals that E. histolytica prevalence may be lower than estimated. PMID- 21618184 TI - Antiparasitic efficiency of Artemisia absinthium on Toxocara cati in naturally infected cats. AB - OBJECTIVE: The first aim of the present study was to determine the efficiency of A. absinthium extract on cats naturally infected with Toxocara cati. The second aim was to determine the efficiency of the extract on the embryonic development of T. cati eggs in vitro. METHODS: Artemisia absinthium extract was orally administrated to cats at the doses of 300 mg/kg and 600 mg/kg body weight in Group 1 and 2, respectively. It was given only once a day and the treatment continued 7 consecutive days. The faeces of the cats were examined both macroscopically and microscopically by flotation procedure with saturated salt solution pre-, during and post- treatment period. The faecal analysis was maintained during 8 days after completing the extract administration. The alteration of faecal egg numbers was performed by using the McMaster technique. RESULTS: The faecal egg numbers per gram were decreased gradually in cats in the trial groups. In the treatment period, the activities of ALT, AST, ALP, urea and creatinine were located within the physiological ranges in cats. In in vitro trials with A. absinthium extract, the embryonic development of T. cati eggs was identical in all groups (treatment and control). A. absinthium extract did not inhibit larval development in eggs in in vitro trials. CONCLUSION: This plant extract may be an alternative choice in the treatment of parasitic diseases in future. PMID- 21618185 TI - [Evaluation of Sabin-Feldman test results of Ankara University Medical Faculty Medical Parasitology laboratory between 1997-2007]. AB - OBJECTIVE: Toxoplasmosis caused by Toxoplasma gondii is a protozoan infection which has a worldwide distribution. Sabin-Feldman is considered as a reference test among serologic methods for its high sensitivity and specificity and has a role for the confirmation of the suspected results of the commonly used ELISA tests. Few laboratories perform this test as it enrolls the live tachyzoites.The aim of this study was to report the results of Ankara University Medical Faculty Parasitology laboratory which is the first lab to perform this test in Turkey. METHODS: We retrospectively investigated anti-Toxoplasma antibody positivities and titers of 678 sera obtained from 648 patients who applied to our laboratory between January 1997 and December 2007 with a prediagnosis of toxoplasmosis. RESULTS: Among 648 patients (490 (75.62%) female and 158 (24.38%) male), 318 (46.90%) were seropositive with titers of 1:16 or higher. The titers were 1:16 in 93, 1:64 in 134, 1:256 in 53, 1:1024 in 28 and 10 patients with higher titers. Seven patients (1.08%) seroconverted or had a 4 fold increase in antibody titers in their sera taken 3 weeks apart and were considered as acute toxoplasmosis. There was no statistically significant difference between the seropositivity rates of female and male patients (p=0.078). CONCLUSION: Sabin-Feldman Test still has an important role in seroprevalence studies both in humans and animals and confirmation of the tests used in routine diagnosis. PMID- 21618186 TI - [Isolation of Blastocystis spp. from human hosts and in vitro determination of different morphological forms]. AB - OBJECTIVE: Blastocystis is a highly common parasite that infects the gastrointestinal tract of many different organisms. Morphology and the appropriate classification of Blastocystis spp. has only recently been resolved with molecular biological studies. This study was performed to determine the prevalence of Blastocystis spp. among humans and to isolate the parasite from clinical specimens. METHODS: Blastocystis spp. was detected in 0.48% of the stool samples and the positive samples were cultivated in Locke-Egg Serum (LES). During passages inoculums were investigated by direct microscopy and stained with trichrome and iron hematoxylene. RESULTS: Vacuolar and granular forms were the most common in cultures and also the amoeboid form was observed. CONCLUSION: LES medium may be a suitable selection for studies aiming to determine the frequency of Blastocystis spp. and for the diagnosis in routine laboratories. PMID- 21618187 TI - Prevalence of Toxoplasma gondii antibodies in domestic (Columba livia domestica) and wild (Columba livia livia) pigeons in Nigde region, Turkey. AB - OBJECTIVE: The present study was conducted to investigate the prevalence of Toxoplasma gondii specific antibodies in domestic (Columba livia domestica) and wild (Columba livia livia) pigeons between October 2003-June 2004. METHODS: Blood samples were collected from 216 pigeons, consisting of 105 (55 female, 50 male) domestic pigeons and 111 (53 female, 58 male) wild pigeons. The sera were tested for T. gondii antibodies using the Sabin Feldman Dye Test (SFDT). RESULTS: One of the 105 (0.95%) domestic pigeon and one of the 111 (0.90%) wild pigeon were found to be seropositive for T. gondii antibodies at the titer of 1:16. CONCLUSION: This is the first serological study on toxoplasmosis in the domestic and wild pigeon in the Nigde region of Turkey. PMID- 21618188 TI - Nematocystis vinodae n. sp. (Protozoa, Apicomplexa, Eugregarinida), a monocystid gregarine from Eutyphoeus nicholsoni (Beddard). AB - OBJECTIVE: Biodiversity studies in search of endoparasitic acephaline gregarines revealed a new species under the genus Nematocystis Hesse, 1909. METHODS: The species has been obtained from the seminal vesicles of the earthworm, Eutyphoeus nicholsoni (Beddard). RESULTS AND CONCLUSION: Gamonts of this species are solitary and bear a perfect nematoid appearance with smoothly curved terminals, especially in mature forms. Mature gamonts measure 245.40-302.66 (270.68+/-18.86) MUm in length and 20.45-26.58 (24.54+/-2.10) MUm in width. Nucleus is rounded or slightly ovoidal, 20.45-24.54 (22.69+/-1.74) MUm in diameter. Gametocysts are almost rounded, measuring 77.71-85.89 (82.82+/-2.69) MUm in diameter. Oocysts are navicular, measuring 7.70-8.47 (8.24+/-0.36) MUm x 4.23-4.62 (4.50 +/-0.18) MUm. PMID- 21618189 TI - [Frequency of intestinal parasites among administrators and workers in sanitary and non-sanitary institutions]. AB - OBJECTIVE: Transmission of parasites generally occurs through fecal-oral means directly from human to human or through receiving eggs and cysts by means of nourishment. The aim of the study was to investigate the frequency of intestinal parasites among administrators and workers in sanitary and non-sanitary institutions. METHODS: Stool specimens were examined using native-lugol, Trichrome and acid-fast stains methods. RESULTS: 23.7% of the 2443 fecal specimens were found to be positive. The frequencies of parasites were found to be 9.8% for Entamoeba coli, 7.2% for Blastocystis hominis, 7.2% for Iodamoeba butschlii, 3.4% for Giardia intestinalis, 0.9% for Dientamoeba fragilis, 0.13% for Entamoeba histolytica, 0.08% for Chilomastix mesnilii, 0.04% for Trichomonas intestinalis, 0.04% for Entamoeba hartmanni, 0.04% for Hymenolepis nana, 0.04% for Taenia spp. and 0.04% for Enterobius vermicularis. CONCLUSION: This rate of parasite positivity among healthy subjects visiting hospital for porter examination suggests that intestinal parasites still constitute a public health problem in the region. Moreover, it can be considered that one important factor in the frequency of the parasite can be both the nature of the jobs of administrators and workers in sanitary and non-sanitary institutions and their interaction with people during sales. PMID- 21618190 TI - Chewing lice (Phthiraptera) found on songbirds (Passeriformes) in Turkey. AB - OBJECTIVE: This study was performed to detect chewing lice species found on the songbirds at Lake Kuyucuk bird ringing station in the Kars province located in eastern Turkey. METHODS: Chewing lice were collected from songbirds captured between September and October 2009. Fifty-one birds belonging to 22 species and 16 genera from 10 families were examined for the louse. RESULTS: Eleven of 51 birds (21.57%) belonging to 7 species; were infested with at least one chewing louse species. The collected lice were identified as Menacanthus chrysophaeus (Kellogg, 1896) on Reed Bunting, Menacanthus pusillus (Nitzsch,1866) on Water Pipits, Calandra Lark and Yellow Wagtail, Myrsidea rustica (Giebel,1874) on Swallow, Brueelia cruciata (Burmeister,1838) on Red-backed Shrike, and Penenirmus rarus (Zlotorzycka,1976) on Chiffchaff. All four Reed Bunting specimens were infested with Menacanthus chrysophaeus. The rate of infestation was 100% in Reed Bunting, Red-backed Shrike and Swallow; 66.7% in Yellow Wagtail; 50% in Calandra Lark and Chiffchaff and 11.1% in Water Pipits. No louse infestation was found in the birds belonging to Paridae, Passeridae, Sylviidae, and Muscipapidae families. CONCLUSION: Menacanthus chrysophaeus on Reed Bunting and Menacanthus pusillus on Calandra Lark are new hosts for these lice species. All the louse species determined in the present study are first records for Turkey. PMID- 21618191 TI - [Some biological features of Hyalomma marginatum in the laboratory]. AB - OBJECTIVE: This study has been planned to investigate some biological features of Hyalomma marginatum, which is one of the important vectors in the world and in our country. METHODS: The study started with the adult H. marginatum which were collected from cattle in the field and attempts were made for these to be adapted to the laboratory by breeding in the laboratory throughout a generation. Unfed adult ticks and unfed larvae were bred on the rabbit ear while unfed nymphs were bred on the calf ear. The biologic development stages of this tick kind, except for sucking blood stage, passed in an incubator which was at a temperature of 28 degrees C and 85% relative humidity, while the unfed active stages passed in an incubator which was at 18 degrees C and 85% relative humidity to conserve for a long time. RESULTS: Unfed females on rabbits were full and fell in approximately 15 days, began to ovulate 20.5 days later and ovulated for 16 days. The larvae which hatched out approximately 29 days later became active in 8.5 days. These larvae sucked blood from rabbits for 14.5 days and abandoned these animals as full nymphs. Unfed adult ticks, which existed as a result of these nymphs casting off their skin in approximately 26 days, became active by completing their cutinization in approximately 10 days. CONCLUSION: At the end of the study, it was determined that H. marginatum showed 2 different behaviors in rabbits. According to this, the total life cycle in the laboratory environment changed by between 97 and 182 days and lasted approximately 138.5 days. PMID- 21618192 TI - [A postoperative wound myiasis caused by Lucilia sericata (Diptera: Calliphoridae) in a woman in Kars]. AB - Lucilia sericata is known as a facultative ectoparasite and it often causes myiasis in open wounds, mouth, nose and eyes of animals and rarely in humans. This case report concerns a 56 year old woman who had right breast cancer surgery. Approximately 10 months after surgery, swelling, pain, redness occurred in the incision site. About two weeks later the site turned into an open wound then fly larvae (97 live larvae) were found. Under t operation conditions, a large surgical wound debridement and debulking were performed on the open wound. The patient was followed for three weeks after the operation. The wound dressings were carried out daily and it shrank partially. Medical treatment was given to the patient and she was discharged. The collected larvae were identified as Lucilia sericata by microscopic examination in the parasitology lab. PMID- 21618193 TI - A nasopharyngeal human infestation caused by Linguatula serrata nymphs in Van province: a case report. AB - The reason for the presentation of this case is that Linguatula serrata, a parasite rarely encountered in humans, was found in a patient in Van province. The patient, who was 26 years old, lived in Ercis a town in Van province, East Turkey. She was admitted to the Outpatient Clinic of Infectious Diseases of Ercis Government Hospital with a complaint of coughing a few worms about 4 cm long from the oral cavity, and also sore throat, partial voice loss and vomiting. The polyclinic doctor suspected that the worm was a parasite but he could not make a diagnosis, and the patient was referred to Health Research and Training Hospital, Yuzuncu Yil University. The parasite was examined in the Parasitology Laboratory and it was observed that this parasite was the nymph of L. serrata whose mouth was surrounded with four hooks and had approximately 90 body segments with small hooks. No medical treatment was given to the patient except that gargling with saline solution was recommended. As a result, we think that physicians should consider L. serrata infestation in patients applying to health foundations with complaints such as pharyngitis accompanied by pharyngeal pain, coughing, sneezing and vomiting. PMID- 21618194 TI - Primary hydatid disease of the pancreas mimicking pancreatic pseudo-cyst in a child: case report and review of the literature. AB - Primary hydatid disease of the pancreas is very rare. We report the case of a 7 year-old girl who presented with abdominal pain and an epigastric mass. The Casoni and indirect hemagglutination test for hydatid disease were negative. A diagnosis of a pancreatic pseudocyst was established by ultrasonography (US) and computed tomography scan before surgery. Ultrasound guided percutaneous drainage was planned as treatment. During the procedure, the cyst was perforated and as germinative membrane was seen by US, we arranged surgery. Hydatid disease should be considered in the differential diagnosis of all cystic masses in the pancreas, even if Casoni and indirect hemagglutination tests negative, especially in geographic regions like Turkey, where the disease is endemic. PMID- 21618195 TI - [Cystic echinococcosis mimicking tuberculosis in childhood]. AB - Both cystic echinococcosis (CE) and tuberculosis (Tbc) are important health problems in developing countries. Pulmonary CE and Tbc have to be differentiated from other diseases as they have increased the risk of morbidity and mortality. Besides, these two diseases can mimic each other. Here, we discuss a 7 year-old patient admitted with fever, cough who was unresponsive to nonspecific antibiotic treatment given for pneumonia, had Tbc treatment due to a positive tuberculin skin test and radiologic appearance consistent with Tbc and on follow-up. He was eventually diagnosed as CE based on a cystic lesion consistent with CE in the liver and echinococcosis specific IgE positivity and was succesfully treated with anti-parasitic therapy. PMID- 21618196 TI - [General overview of camel parasites and the situation in Turkey]. AB - The aim of this review is overview the main protozoan, arthropoda and helminthic parasites seen in camels and to evaluate the publications related to camel parasites in Turkey. In different parts of the world, trichostrongylose, mange, nasal myiasis and trypanosomiasis are the most common parasitic diseases found in camels. Hydatic cyst larval stage of E. granulosus is important both economically and in terms of public health. It is also emphasized that the records concerning parasites of the Turkish camel population, which is decreasing significantly, are extremely limited. PMID- 21618197 TI - Clinical images: Polyostotic fibrous dysplasia. PMID- 21618198 TI - Increased sensitivity to apoptosis induced by methotrexate is mediated by JNK. AB - OBJECTIVE: Low-dose methotrexate (MTX) is an effective therapy for rheumatoid arthritis (RA), yet its mechanism of action is incompletely understood. The aim of this study was to explore the induction of apoptosis by MTX. METHODS: Flow cytometry was performed to assess changes in the levels of intracellular proteins, reactive oxygen species (ROS), and apoptosis. Quantitative polymerase chain reaction was performed to assess changes in the transcript levels of select target genes in response to MTX. RESULTS: MTX did not directly induce apoptosis but rather "primed" cells for markedly increased sensitivity to apoptosis via either mitochondrial or death receptor pathways, by a JNK-dependent mechanism. Increased sensitivity to apoptosis was mediated, at least in part, by MTX dependent production of ROS, JNK activation, and JNK-dependent induction of genes whose protein products promote apoptosis. Supplementation with tetrahydrobiopterin blocked these MTX-induced effects. Patients with RA who were receiving low-dose MTX therapy expressed elevated levels of the JNK target gene, jun. CONCLUSION: Our results support a model whereby MTX inhibits reduction of dihydrobiopterin to tetrahydrobiopterin, resulting in increased production of ROS, increased JNK activity, and increased sensitivity to apoptosis. The finding of increased jun levels in patients with RA receiving low-dose MTX supports the notion that this pathway is activated by MTX in vivo and may contribute to the efficacy of MTX in inflammatory disease. PMID- 21618200 TI - A possible role of leptin-associated increase in soluble interleukin-2 receptor diminishing a clinical response to infliximab in rheumatoid arthritis: Comment on the article by Klaasen et al. PMID- 21618201 TI - Subcutaneous abatacept versus intravenous abatacept: a phase IIIb noninferiority study in patients with an inadequate response to methotrexate. AB - OBJECTIVE: To compare the efficacy and safety of subcutaneous (SC) and intravenous (IV) abatacept. METHODS: In this phase IIIb double-blind, double dummy, 6-month study, patients with rheumatoid arthritis (RA) and inadequate responses to methotrexate were randomized to receive 125 mg SC abatacept on days 1 and 8 and weekly thereafter (plus an IV loading dose [~10 mg/kg] on day 1) or IV abatacept (~10 mg/kg) on days 1, 15, and 29 and every 4 weeks thereafter. The primary end point for determining the noninferiority of SC abatacept to IV abatacept was the proportion of patients in each group meeting the American College of Rheumatology 20% improvement criteria (achieving an ACR20 response) at month 6. Other efficacy end points, immunogenicity, and safety were also assessed. RESULTS: Of 1,457 patients, 693 of 736 (94.2%) treated with SC abatacept and 676 of 721 (93.8%) treated with IV abatacept completed 6 months. At month 6, 76.0% (95% confidence interval 72.9, 79.2) of SC abatacept-treated patients versus 75.8% (95% confidence interval 72.6, 79.0) of IV abatacept treated patients achieved an ACR20 response (estimated difference between groups 0.3% [95% confidence interval -4.2, 4.8]), confirming noninferiority of SC abatacept to IV abatacept. Onset and magnitude of ACR responses and disease activity and physical function improvements were comparable between the SC and IV abatacept-treated groups. The proportions of adverse events (AEs) and serious AEs over 6 months were 67.0% and 4.2%, respectively, in the SC abatacept-treated group and 65.2% and 4.9%, respectively, in the IV abatacept-treated group, with comparable frequencies of serious infections, malignancies, and autoimmune events between groups. SC injection site reactions (mostly mild) occurred in 19 SC abatacept (IV placebo)-treated patients (2.6%) and 18 IV abatacept (SC placebo) treated patients (2.5%). Abatacept-induced antibodies occurred in 1.1% of SC abatacept-treated patients and 2.3% of IV abatacept-treated patients. CONCLUSION: SC abatacept provides efficacy and safety comparable with that of IV abatacept, with low immunogenicity and high retention rates, consistent with the established IV abatacept profile. Rates of injection site reactions were low. SC abatacept will provide additional treatment options, such as an alternative route of administration, for patients with RA. PMID- 21618202 TI - Association of the idiotype:antiidiotype antibody ratio with the efficacy of intravenous immunoglobulin treatment for the prevention of recurrent autoimmune associated congenital heart block. AB - OBJECTIVE: Congenital heart block (CHB), a manifestation of neonatal lupus, is associated with maternal anti-Ro/SSA and anti-La/SSB autoantibodies and recurs in ~18% of subsequent pregnancies. This study was undertaken to investigate the effect of the idiotype:antiidiotype (Id:anti-Id) antibody ratio in the ability of intravenous immunoglobulin (IVIG) administered during subsequent pregnancies to prevent CHB. METHODS: We studied 16 anti-Ro/SSA and anti-La/SSB-positive pregnant women from the Preventive IVIG Therapy for Congenital Heart Block study who had previously given birth to a child with neonatal lupus. In 3 of the mothers, the study pregnancy resulted in the birth of a child with neonatal lupus (2 with CHB and 1 with rash). Sequential serum samples were obtained from all mothers immediately before the administration of IVIG during pregnancy and were evaluated for antibodies against the major B cell epitope 349-364aa of La/SSB (idiotype) and its antiidiotypic antibodies. RESULTS: Following IVIG treatment, serum titers of anti-La(349-364) (Id antibodies) decreased in 80% of the mothers, and in 60% an increase in anti-Id antibodies against anti-La(349-364) was observed. The Id:anti-Id ratio was significantly higher in mothers whose offspring developed neonatal lupus compared to mothers who gave birth to a healthy child (P<0.0001). Removal of anti-Id antibodies substantially increased the reactivity against La(349-364) in sera from 5 of 7 mothers tested. All IVIG preparations were examined for Id and anti-Id antibody activity. IVIG from batches administered to mothers who gave birth to a healthy child had an Id:anti-Id activity ratio of <1, in contrast to that given to mothers who gave birth to a child with neonatal lupus. Addition of the IVIG preparations to the maternal sera further enhanced antiidiotypic activity (by up to 4.7-fold) in 11 of 13 patients studied. CONCLUSION: This is the first study in humans to demonstrate that IVIG influences the Id-anti-Id network of a specific pathogenic autoantibody. Specifically, we showed that IVIG enhanced the anti-Id antibody response in pregnant women with anti-La/SSB antibodies. A high Id:anti-Id ratio in both the IVIG preparation and the maternal serum may explain the absence of an effect of IVIG in preventing recurrent neonatal lupus in some cases. PMID- 21618203 TI - Chitinases in the salivary glands and circulation of patients with Sjogren's syndrome: macrophage harbingers of disease severity. AB - OBJECTIVE: Sjogren's syndrome (SS) is a chronic autoimmune disease of unknown etiology that targets salivary and lacrimal glands and may be accompanied by multiorgan systemic manifestations. To further the understanding of immunopathology associated with SS and identify potential therapeutic targets, we undertook the present study comparing the gene expression profiles of salivary glands with severe inflammation versus those of salivary glands with mild or no disease. METHODS: Using microarray profiling of salivary gland tissue from patients with SS and control subjects, we identified target genes, which were further characterized in tissue, serum, and cultured cell populations by real time polymerase chain reaction and protein analysis. RESULTS: Among the most highly expressed SS genes were those associated with myeloid cells, including members of the mammalian chitinase family, which had not previously been shown to be associated with exocrinopathies. Both chitinase 3-like protein 1 and chitinase 1, highly conserved chitinase-like glycoproteins (one with enzymatic activity and one lacking enzymatic activity), were evident at the transcriptome level and were detected within inflamed tissue. Chitinases were expressed during monocyte-to macrophage differentiation and their levels augmented by stimulation with cytokines, including interferon-alpha (IFNalpha). CONCLUSION: Because elevated expression of these and other macrophage-derived molecules corresponded with more severe SS, the present observations suggest that macrophages have potential immunopathologic involvement in SS and that the tissue macrophage transcription profile reflects multiple genes induced by IFNalpha. PMID- 21618204 TI - B cell biomarkers of rituximab responses in systemic lupus erythematosus. AB - OBJECTIVE: Rituximab appears to be effective in many studies of systemic lupus erythematosus (SLE), with variable initial clinical response and time to relapse. However, results of a randomized controlled trial of rituximab were negative. This study was undertaken to evaluate the effectiveness of rituximab in SLE, using highly sensitive flow cytometry (HSFC), which can define B cell numbers 50 100 times lower than conventional techniques and predicts responses in rheumatoid arthritis. METHODS: Thirty-nine patients with active SLE were started on a standard regimen of rituximab with intravenous and oral steroids. Clinical response and relapse were defined using the British Isles Lupus Assessment Group (BILAG) index with criteria for major clinical response, partial clinical response, and nonresponse. HSFC, including analysis of B cell subsets, was performed. RESULTS: There was a significant reduction from baseline in global BILAG score at all time points analyzed (P<0.0001), and major clinical response and partial clinical response rates were 51% and 31%, respectively. Time to relapse was highly variable. Fifty percent of the patients relapsed after 6-18 months (earlier relapse); the remainder relapsed at a slower rate (later relapse). B cell depletion and repopulation were variable and were predictive of these clinical outcomes. There was a persistent B cell presence in 21 patients after 2 infusions of rituximab, which included all 7 patients with no response (P=0.012 versus patients with complete depletion of B cells). Memory B cell (P=0.02) and plasmablast (P<0.001) repopulation after 26 weeks was markedly faster in patients with earlier relapse versus patients with later relapse. CONCLUSION: Our findings indicate that rituximab is effective in SLE, and clinical responses are supported by close correlation with B cell numbers. HSFC is a valuable tool in the assessment and prediction of response in SLE. PMID- 21618205 TI - Clinical course of lung physiology in patients with scleroderma and interstitial lung disease: analysis of the Scleroderma Lung Study Placebo Group. AB - OBJECTIVE: Patients with systemic sclerosis-associated interstitial lung disease (SSc-ILD) are thought to have the greatest decline in lung function (forced vital capacity [FVC]% predicted) in the early years after disease onset. The aim of this study was to assess the natural history of the decline in FVC% predicted in patients receiving placebo in the Scleroderma Lung Study and to evaluate possible factors for cohort enrichment in future therapeutic trials. METHODS: Patients randomized to receive placebo (n=79) were divided into 3 groups based on the duration of SSc (0-2 years, 2-4 years, and >4 years). Descriptive statistics and a mixed-effects model were used to analyze the rate of decline in the FVC% predicted over a 1-year period. Additional analyses stratified according to the severity of fibrosis on high-resolution computed tomography (HRCT) were performed, and interactions between disease severity and disease duration were explored. RESULTS: The mean+/-SD decline in the unadjusted FVC% predicted during the 1-year period was 4.2+/-12.8%. At baseline, 28.5%, 43.0%, and 28.5% of patients were in the groups with disease durations of 0-2 years, 2-4 years, and >4 years, respectively. The rate of decline in the FVC% predicted was not significantly different across the 3 disease groups (P=0.85). When stratified by baseline fibrosis on HRCT, the rate of decline in the FVC% predicted was statistically significantly greater in the group with severe fibrosis (mean annualized decline in the FVC% predicted 7.2% versus 2.7% in the groups with no or moderate fibrosis; P=0.008). The decline observed in the group with severe fibrosis was most pronounced in those with a relatively short disease duration (0 2 years; annualized decline 7.0%). CONCLUSION: Among patients with SSc-ILD in the Scleroderma Lung Study, the rates of progression of lung disease were similar irrespective of disease duration. The baseline HRCT fibrosis score is a predictor of a future decline in the FVC% predicted in the absence of effective treatment. PMID- 21618206 TI - Transforming growth factor beta controls CCN3 expression in nucleus pulposus cells of the intervertebral disc. AB - OBJECTIVE: To investigate transforming growth factor beta (TGFbeta) regulation of CCN3 expression in cells of the nucleus pulposus. METHODS: Real-time reverse transcription-polymerase chain reaction and Western blot analyses were used to measure CCN3 expression in the nucleus pulposus. Transfections were used to measure the effect of Smad3, MAPKs, and activator protein 1 (AP-1) on TGFbeta mediated CCN3 promoter activity. Lentiviral knockdown of Smad3 was performed to assess the role of Smad3 in CCN3 expression. RESULTS: CCN3 was expressed in embryonic and adult intervertebral discs. TGFbeta decreased the expression of CCN3 and suppressed its promoter activity in nucleus pulposus cells. DN-Smad3, Smad3 small interfering RNA, or DN-AP-1 had little effect on TGFbeta suppression of CCN3 promoter activity. However, p38 and ERK inhibitors blocked suppression of CCN3 by TGFbeta, suggesting involvement of these signaling pathways in the regulation of CCN3. Interestingly, overexpression of Smad3 in the absence of TGFbeta increased CCN3 promoter activity. We validated the role of Smad3 in controlling CCN3 expression in Smad3-null mice and in nucleus pulposus cells transduced with lentiviral short hairpin Smad3. In terms of function, treatment with recombinant CCN3 showed a dose-dependent decrease in the proliferation of nucleus pulposus cells. Moreover, CCN3-treated cells showed a decrease in aggrecan, versican, CCN2, and type I collagen expression. CONCLUSION: The opposing effect of TGFbeta on CCN2 and CCN3 expression and the suppression of CCN2 by CCN3 in nucleus pulposus cells further the paradigm that these CCN proteins form an interacting triad, which is possibly important in maintaining extracellular matrix homeostasis and cell numbers. PMID- 21618207 TI - Increased expression of interleukin-22 by synovial Th17 cells during late stages of murine experimental arthritis is controlled by interleukin-1 and enhances bone degradation. AB - OBJECTIVE: Interleukin-22 (IL-22) is a mediator in antimicrobial responses and inflammatory autoimmune diseases. Although IL-22 and its receptor, IL-22R, have been identified in the synovium of rheumatoid arthritis patients, the source of IL-22 and its contribution to disease pathogenicity remain to be established. This study was undertaken to investigate the regulation of IL-22 by Th17 cells in vitro and to evaluate the potential for IL-22 depletion in an experimental arthritis model using mice deficient in the IL-1 receptor antagonist (IL-1Ra-/-). METHODS: Naive murine T cells were cultured under conditions leading to polarization of the cells into subsets of Th1, Th2, induced Treg, and Th17. Cytokines were measured in the culture supernatants, and the cells were analyzed by fluorescence-activated cell sorting. Tissue samples from the inflamed ankle synovium of IL-1Ra-/- mice were isolated, and messenger RNA levels of marker genes were quantified. IL-1Ra-/- mice were treated with neutralizing anti-IL-22 antibodies. Synovial cells were isolated from the inflamed tissue and sorted into fractions for analysis of cytokine production. RESULTS: In vitro tests showed that Th17 cells produced high levels of IL-22 after stimulation with IL-1 or IL 23. Interestingly, a synergistic increase in the production of IL-22 was observed after combining IL-1 and IL-23. In vivo, IL-1Ra-/- mice displayed a progressive erosive arthritis, characterized by up-regulation of IL-17 in mildly and severely inflamed tissue, whereas the levels of IL-22 and IL-22R were increased only in severely inflamed synovia. Anti-IL-22 treatment of IL-1Ra-/- mice significantly reduced the inflammation and bone erosion. Analysis of isolated single cells from the inflamed synovia revealed that IL-22 was mainly produced by IL-17-expressing T cells. CONCLUSION: These findings suggest that IL-22 plays an important role in IL-1-driven chronic joint destruction. PMID- 21618208 TI - The specific free radical scavenger edaravone suppresses fibrosis in the bleomycin-induced and tight skin mouse models of systemic sclerosis. AB - OBJECTIVE: Patients with systemic sclerosis (SSc) exhibit enhanced production of free radicals due to ischemia and reperfusion injury following Raynaud's phenomenon, an initial clinical manifestation. Oxidative stress induces cytokine production, inflammatory cell recruitment, and tissue injury in several inflammatory diseases. The aim of this study was to examine the effect of edaravone, a free radical scavenger, on the development of fibrosis and autoimmunity in two different mouse models of SSc. METHODS: The bleomycin-induced SSc model in mice and the tight skin mouse model were used to evaluate the effect of edaravone on fibrosis and immunologic abnormalities. To assess the reaction of fibroblasts to stimulation with free radicals, fibroblasts from these mice were cultured with NONOate, a nitric oxide-releasing agent, and hydrogen peroxide. RESULTS: Treatment with edaravone reduced fibrosis in mice with bleomycin-induced SSc and in TSK/+ mice. The production of free radicals was also attenuated by edaravone in both models. In addition, production of fibrogenic cytokines such as interleukin-6 and transforming growth factor beta1, production of anti topoisomerase I antibody, and the degree of hypergammaglobulinemia were reduced by edaravone. Furthermore, bleomycin induced the production of H2O2 and nitric oxide from inflammatory cells, and collagen production was increased in fibroblasts cultured with H2O2 and NONOate. CONCLUSION: This study is the first to show that edaravone has a significant inhibitory effect on fibrosis both in the bleomycin-induced SSc model and in TSK/+ mice. These results indicate that edaravone should be further evaluated for potential use as an antifibrotic agent in SSc. PMID- 21618209 TI - Zinc to cadmium replacement in the A. thaliana SUPERMAN Cys2 His2 zinc finger induces structural rearrangements of typical DNA base determinant positions. AB - Among heavy metals, whose toxicity cause a steadily increasing of environmental pollution, cadmium is of special concern due to its relatively high mobility in soils and potential toxicity at low concentrations. Given their ubiquitous role, zinc fingers domains have been proposed as mediators for the toxic and carcinogenic effects exerted by xenobiotic metals. To verify the structural effects of zinc replacement by cadmium in zinc fingers, we have determined the high resolution structure of the single Cys2 His2 zinc finger of the Arabidopsis thaliana SUPERMAN protein (SUP37) complexed to the cadmium ion by means of UV-vis and NMR techniques. SUP37 is able to bind Cd(II), though with a dissociation constant higher than that measured for Zn(II). Cd-SUP37 retains the betabetaalpha fold but experiences a global structural rearrangement affecting both the relative orientation of the secondary structure elements and the position of side chains involved in DNA recognition: among them Ser17 side chain, which we show to be essential for DNA binding, experiences the largest displacement. PMID- 21618210 TI - Yield stress of pretreated corn stover suspensions using magnetic resonance imaging. AB - Cellulose fibers in water form networks that give rise to an apparent yield stress, especially at high solids contents. Measuring the yield stress and correlating it with fiber concentration is important for the biomass and pulp industries. Understanding how the yield stress behaves at high solids concentrations is critical to optimize enzymatic hydrolysis of biomass in the production of biofuels. Rheological studies on pretreated corn stover and various pulp fibers have shown that yield stress values correlate with fiber mass concentration through a power-law relationship. We use magnetic resonance imaging (MRI) as an in-line rheometer to measure velocity profiles during pipe flow. If coupled with pressure drop measurements, these allow yield stress values to be determined. We compare our results with literature values and discuss the accuracy and precision of the rheo-MRI measurement, along with the effects of fiber characteristics on the power-law coefficients. PMID- 21618211 TI - Trends and outcomes after surgical lower limb revascularization in England. AB - BACKGROUND: The aim was to analyse contemporary data on the number of surgical revascularization procedures performed each year in England, and their outcome. METHODS: Hospital Episode Statistics and Office for National Statistics data were used to quantify numbers and identify factors associated with outcome after all femoropopliteal and femorodistal bypass procedures performed between 2002 and 2006. Outcome measures were repeat bypass, major amputation, death and a composite measure. Single-level multivariable logistic regression modelling was used to quantify the effect of these variables on outcome. RESULTS: A total of 21,675 femoropopliteal and 3458 femorodistal bypass procedures were performed. Mean in-hospital mortality rates were 6.7 and 8.0 per cent respectively. One-year survival rates were 82.8 and 79.1 per cent; both increased over the study interval. The mean 1-year major amputation rate after femoropopliteal bypass was 10.4 per cent, which decreased significantly over the 5 years (P < 0.001); after distal bypass the rate of 20.8 per cent remained unchanged (P = 0.456). Diabetes mellitus and chronic kidney disease were significant predictors of adverse outcome for both procedures: odds ratio (OR) at 1 year 1.56 (95 per cent confidence interval 1.46 to 1.67; P < 0.001) and 2.15 (1.88 to 2.45; P < 0.001) respectively for femoropopliteal bypass. Previous femoral angioplasty was associated with an increased rate of major amputation 1 year after proximal bypass (OR 1.18, 1.05 to 1.33; P = 0.004). CONCLUSION: Although all mortality rates are improving, the major amputation rate remains high after femorodistal bypass. Adverse events occurred after 37.6 per cent of femoropopliteal and 49.7 per cent of femorodistal bypasses; diabetes and chronic renal failure were the main predictors of poor outcome. PMID- 21618212 TI - Risk factors for conversion and complications after unilateral laparoscopic adrenalectomy. AB - BACKGROUND: Laparoscopic adrenalectomy (LA) is the procedure of choice for surgical management of most benign adrenal tumours, with a reported overall complication rate around 10 per cent. The aim of this study was to determine predictive factors for postoperative complications and conversion to open surgery after unilateral LA. METHODS: From 1994 to 2009, consecutive patients undergoing unilateral LA by the lateral transabdominal approach were analysed from a prospectively maintained database. A mass larger than 12 cm in diameter and suspected primary adrenal carcinoma were considered contraindications to LA. Predictive factors for postoperative complications and conversion to open surgery were analysed. RESULTS: Some 462 patients were analysed. There were no postoperative deaths. Postoperative complications occurred in 53 patients (11.5 per cent), medical complications in 28, and surgical complications in 33 patients. Six patients underwent reoperation for complications. Multivariable logistic regression analysis showed that conversion to open surgery (odds ratio (OR) 6.20, 95 per cent confidence interval 2.08 to 18.53; P = 0.001) and left sided tumour (OR 1.89, 1.02 to 3.52; P = 0.044) were independent predictive factors for overall complications. Conversion to open surgery was the only independent predictive factor for medical complications (OR 12.88, 4.21 to 39.41; P = 0.001), and left-sided LA was the only predictive factor for surgical complications (OR 2.22, 1.01 to 4.89; P = 0.047). No factor was predictive of conversion to open surgery. CONCLUSION: In this single-institution study, conversion to open surgery and left-sided tumours were independent predictive factors for overall complications, but none of the variables analysed was predictive of conversion. PMID- 21618213 TI - HER-2 status in primary oesophageal cancer, lymph nodes and distant metastases. AB - BACKGROUND: Some 10-15 per cent of patients with oesophageal cancer overexpress human epidermal growth factor receptor (HER) 2 at the primary tumour site, leading to the hope that specific targeted systemic therapy might favourably influence clinical and subclinical disease at locoregional and distant sites. This approach is based on primary tumour characteristics, without knowledge of expression patterns at metastatic sites. In oesophageal cancer, concordance between HER-2 status at the primary tumour and other sites is unknown. METHODS: The HER-2 status of primary tumours and corresponding metastatic sites (lymph node and distant) and local recurrence were evaluated in a series of patients with oesophageal cancer, using immunohistochemistry and dual colorimetric in situ hybridization. RESULTS: There were 97 adenocarcinomas (ACs) and 79 squamous cell carcinomas (SCCs). Some 14 per cent of primary ACs and 1 per cent of primary SCCs were staged as HER-2-positive. The HER-2 status was identical in the primary tumour and lymph node metastases in 95 per cent of ACs and 99 per cent of SCCs respectively (P = 0.375, sign test). Nineteen of 22 distant metastases from AC and all from SCC had identical HER-2 status to the primary tumour. In two of 22 patients with AC the primary tumour was classed as negative but distant metastases were HER-2-positive. CONCLUSION: With over 85 per cent concordance in HER-2 status between primary tumours and distant metastases in oesophageal cancer, routine HER-2 testing of metastases to confirm HER-2 positivity is not warranted. Assessment of HER-2 status at metastatic sites may be worthwhile in some patients with easily accessible metastases and negative HER-2 status at the primary tumour, or if adequate material cannot be obtained from the primary site. PMID- 21618214 TI - Effect of time errors on infinity values obtained using Prony's method. AB - The estimates of infinity values obtained using Prony's method are shown to be subject to significant error arising from small errors in the assigned sample times. The error of the estimate may be reduced by increasing the time interval between samples. PMID- 21618215 TI - Synthesis and antitumor activity of 4-[p-[bis(2 chloroethyl)amino]phenyl]butyrates. AB - Ten 4-[p-[bis(2-chloroethyl)amino]phenyl]butyrates were synthesized and evaluated for antitumor activity. The 2-phenoxyethyl ester exhibited activity against P-388 lymphocytic leukemia, and the n-butyl and n-pentyl esters exhibited activity against L-1210 lymphoid leukemia in initial screening tests. PMID- 21618216 TI - MicroRNA miR-335 is crucial for the BRCA1 regulatory cascade in breast cancer development. AB - The expression of microRNAs is altered in various cancer types, leading to their definition as onco- and tumor-suppressor microRNAs. In our study, we investigated the role of miR-335 in the formation of sporadic human breast cancer and its involvement in the regulatory network of the breast cancer susceptibility gene BRCA1. To validate single components of the BRCA1 cascade, microRNA overexpression was performed in a cell culture model with subsequent protein analysis and luciferase reporter assays. Here, we were able to identify miR-335 as simultaneously regulating the known BRCA1 activators ERalpha, IGF1R, SP1 and the repressor ID4, including a feedback regulation of miR-335 expression by estrogens. Overexpression of miR-335 resulted in an upregulation of BRCA1 mRNA expression, suggesting a functional dominance of ID4 signaling. The relevance of the miR-335 regulation for human breast cancer was confirmed in primary sporadic breast cancer specimens with significantly decreased miR-335 levels (p < 0.05) in comparison to normal controls. Interestingly, the microRNA expression level correlated positively to the BRCA1 transcript level, supporting the hypothesis of a miR-335-mediated regulation of the tumor suppressor gene. Functionally, overexpression of miR-335 led to decreased cell viability and an increase in apoptosis, supporting its tumor-suppressive function. In summary, our data indicate that miR-335 affects different targets in the upstream BRCA1-regulatory cascade with impact on key cellular functions such as proliferation and apoptosis. Deregulation of the microRNA during breast cancer development and progression may thereby lead to an increased tumorigenic potential by inactivating crucial tumor-suppressive signals. PMID- 21618217 TI - Monocytes enhance cell proliferation and LMP1 expression of nasal natural killer/T-cell lymphoma cells by cell contact-dependent interaction through membrane-bound IL-15. AB - Nasal natural killer (NK)/T-cell lymphoma (NNKTL) is an Epstein-Barr virus (EBV) related malignancy with poor prognosis and has distinct histological features characterized by angiocentric and polymorphous lymphoreticular infiltrates including inflammatory cells such as granulocytes, monocytes, macrophages and lymphocytes. Here, we show that the monocytes enhance proliferation as well as LMP1 expression of NNKTL cells by cell contact-dependent interaction through membrane-bound interleukin (IL)-15. We used two EBV-positive NK-cell lines, SNK6 and KAI3, which originated from two patients-SNK6 from a patient with NNKTL and KAI3 from a patient with a severe mosquito allergy. We cocultured the cell lines with granulocytes or monocytes and examined whether proliferation, survival and LMP1 expression of the cells changed. Although cocultured granulocytes did not affect proliferation, survival or LMP1 expression of the cells, cocultured monocytes enhanced both proliferation and LMP1 expression in a dose-dependent manner. These phenomena were not seen when monocytes were placed in a separate chamber. Moreover, the monocyte-inducible proliferation and LMP1 expression were inhibited by treatment with an antibody against IL-15. Furthermore, production of interferon-gamma-inducible protein (IP)-10 were enhanced by coculture with monocytes and were inhibited by the antibody. Immunohistological studies confirmed that a number of infiltrating CD14-positive monocytes contacted CD56 positive lymphoma cells in all of 20 NNKTL tissues tested. These results suggest that monocytes enhance cell growth as well as LMP1 expression of NNKTL cells by cell contact-dependent interaction through membrane-bound IL-15. In the microenvironment of NNKTL tissue, a positive feedback loop of interaction between lymphoma cells and monocytes may be present and contribute to lymphoma progression. PMID- 21618218 TI - A reassessment of whether cortical motor neurons die following spinal cord injury. AB - Over the past century, the question of whether the cells of origin of the corticospinal tract (CST) die following spinal cord injury (SCI) has been debated. A recent study reported an approximately 20% loss of retrogradely labeled cortical motoneurons following damage to their axons resulting from SCI at T9 (Hains et al. [2003] J. Comp. Neurol. 462:328-341). In follow-up studies, however, we failed to find any evidence of loss of CST axons in the medullary pyramid, which must occur if CST neurons die. Here, we seek to resolve the discrepancy by re-evaluating possible loss of CST neurons using the same techniques as Hains et al. (quantitative analysis of retrograde labeling and staining for cell death markers including TUNEL and Hoechst labeling of the nuclei). Following either dorsal funiculus lesions at thoracic level 9 (T9) or lateral hemisection at cervical level 5 (C5), our results reveal no evidence for a loss of retrogradely labeled neurons and no evidence for TUNEL staining of axotomized cortical motoneurons. These results indicate that CST cell bodies do not undergo retrograde cell death following SCI, and therefore targeting such cell death is not a valid therapeutic target. PMID- 21618219 TI - The mouse olfactory peduncle. AB - The olfactory peduncle, the region connecting the olfactory bulb with the basal forebrain, contains several neural areas that have received relatively little attention. The present work includes studies that provide an overview of the region in the mouse. An analysis of cell soma size in pars principalis (pP) of the anterior olfactory nucleus (AON) revealed considerable differences in tissue organization between mice and rats. An unbiased stereological study of neuron number in the cell-dense regions of pars externa (pE) and pP of the AON of 3-, 12 , and 24-month-old mice indicated that pE has about 16,500 cells in 0.043 mm(3) and pP about 58,300 cells in 0.307 mm(3) . Quantitative Golgi studies of pyramidal neurons in pP suggested that mouse neurons are similar to although smaller than those of the rat. An immunohistochemical analysis demonstrated that all peduncular regions (pE, pP, the dorsal peduncular cortex, ventral tenia tecta, and anterior olfactory tubercle and piriform cortex) have cells that express either calbindin, calretinin, parvalbumin, somatostatin, vasoactive intestinal polypeptide, neuropeptide Y, or cholecystokinin (antigens commonly co expressed by subspecies of gamma-aminobutyric acid [GABA]ergic neurons), although the relative numbers of each cell type differ between zones. Finally, an electron microscopic comparison of the organization of myelinated fibers in lateral olfactory tract in the anterior and posterior peduncle indicated that the region is less orderly in mice than in rats. The results provide a caveat for investigators who generalize data between species, as both similarities and differences between the laboratory mouse and rat were observed. PMID- 21618220 TI - Immunohistochemical localization of vesicular glutamate transporter 2 (vGluT2) in the central nervous system of the pigeon (Columba livia). AB - Glutamatergic neurons are distributed widely in the telencephalic pallium of birds, but their targets are unknown. In the present study, a polyclonal antibody was produced against pigeon vesicular glutamate transporter 2 (vGluT2) and was used for Western blot and immunohistochemistry to detect projection targets of glutamatergic neurons in the pigeon central nervous system. The molecular weight of vGluT2 was measured at 65 kDa. vGluT2 immunoreactivity was observed in neuropil, but not in neuronal cell bodies or glia. Immunoreactive neuropil generally appeared homogeneous, but fine granules or puncta were found in many areas or nuclei. The telencephalon showed strong immunoreactivity, except for the globus pallidus. In particular, the mesopallium and hippocampal formation revealed the most intense immunostaining. In the diencephalon, vGluT2 immunoreactivity was intense in the dorsal thalamus and hypothalamus. In the midbrain, strong immunostaining was seen in the periaqueductal gray, the dorsal part of the lateral mesencephalic nucleus, and the isthmo-optic nucleus. The optic tectum showed moderate immunoreactivity. In the cerebellar cortex, glomeruli in the granular layer were intensely immunoreactive, and the molecular layer showed intensely homogeneous immunostaining. In the caudal brainstem, the cochlear magnocellular and angular nuclei showed strongly immunoreactive puncta around neuronal cell bodies. In the spinal cord, the dorsal horn revealed moderate immunoreactivity and the marginal nucleus was strongly immunoreactive. Ultrastructural observations revealed that vGluT2 immunoreactivity is localized in asymmetric, presynaptic terminals. The present results indicate an extensive distribution of glutamatergic projections and circuits in the avian central nervous system. PMID- 21618221 TI - Analyses of the spatiotemporal expression and subcellular localization of liprin alpha proteins. AB - The members of the Liprin-alpha protein family, Liprin-alpha1-4, are scaffolding proteins that play important roles in the regulation of synapse assembly and maturation, vesicular trafficking, and cell motility. Recent evidence suggests that despite their high degree of homology, the four isoforms can be differentially regulated and fulfill diverging functions. However, to date their precise regional and subcellular distribution has remained elusive. Here, we examine the spatiotemporal expression patterns of Liprins-alpha in the rodent by using in situ hybridization, immunoblotting, and immunochemistry of primary cells as well as brain and retina sections. We show that Liprin-alpha1-4 mRNA and protein are widely expressed throughout the developing and adult central nervous system, with Liprin-alpha2 and -alpha3 being the major Liprin-alpha isoforms in the brain. Our data show that the four Liprin-alpha proteins differ in their regional distribution, in particular in the hippocampus, the cerebellum, and the olfactory bulb. Liprin-alpha1 exhibits a unique spatiotemporal expression pattern as its levels decrease during synaptogenesis, and it is the only Liprin-alpha with substantial non-neuronal expression. Immunocytochemistry of cultured primary neurons with pre- and postsynaptic marker proteins shows all four Liprins-alpha to be present at synapses and nonsynaptic sites to varying degrees. Together, these results show that neurons in different brain regions express a distinct complement of Liprin-alpha proteins. PMID- 21618222 TI - Differential expression of liprin-alpha family proteins in the brain suggests functional diversification. AB - Liprin-alpha proteins are major protein constituents of synapses and are important for the organization of synaptic vesicles and neurotransmitter receptors on their respective sides of the synapse. Although it is becoming apparent that the single liprin-alpha gene in invertebrates is essential for synapse function, it is not known to what extent the four different liprin-alpha homologs (liprin-alpha1-4) in mammals are involved at synapses. We have designed specific antibodies against each of the four liprin-alpha proteins and investigated their regional and cellular distribution in the brain. Here we show that all four liprin-alpha proteins are present throughout the mature brain but have different regional distributions, which is highlighted by their differential localization in olfactory bulb, hippocampus, and cerebellar cortex. Double immunofluorescence staining indicates that different liprin-alpha proteins are enriched in different synaptic populations but are also present at nonsynaptic sites. In particular, liprin-alpha2 is preferentially associated with hippocampal mossy fiber endings in the CA3, whereas synapses in the molecular layers of the CA1 and dentate gyrus double-labeled for liprin-alpha3. The localization of liprin-alpha2 and liprin-alpha3 with excitatory synapses was confirmed in cultured primary hippocampal neurons. Liprin-alpha4, which poorly co-distributed with presynaptic markers in hippocampus, instead strongly co-localized with VGLUT1 in the cerebellar molecular layer, suggesting its presence in parallel fiber-Purkinje cell synapses. Finally, staining of cultured glial cells indicated that liprin-alpha1 and liprin-alpha3 are also associated with astrocytes. We conclude that liprin-alpha family proteins might perform independent and specialized synaptic and nonsynaptic functions in different regions of the brain. PMID- 21618223 TI - Galanin neurons in the intermediate nucleus (InM) of the human hypothalamus in relation to sex, age, and gender identity. AB - The intermediate nucleus (InM) in the preoptic area of the human brain, also known as the sexually dimorphic nucleus of the preoptic area (SDN-POA) and the interstitial nucleus of the anterior hypothalamus-1 (INAH-1) is explored here. We investigated its population of galanin-immunoreactive (Gal-Ir) neurons in relation to sex, age, and gender identity in the postmortem brain of 77 subjects. First we compared the InM volume and number of Gal-Ir neurons of 22 males and 22 females in the course of aging. In a second experiment, we compared for the first time the InM volume and the total and Gal-Ir neuron number in 43 subjects with different gender identities: 14 control males (M), 11 control females (F), 10 male-to-female (MtF) transsexual people, and 5 men who were castrated because of prostate cancer (CAS). In the first experiment we found a sex difference in the younger age group (<45 years of age), i.e., a larger volume and Gal-Ir neuron number in males and an age difference, with a decrease in volume and Gal-Ir neuron number in males > 45 years. In the second experiment the MtF transsexual group presented an intermediate value for the total InM neuron number and volume that did not seem different in males and females. Because the CAS group did not have total neuron numbers that were different from the intact males, the change in adult circulating testosterone levels does not seem to explain the intermediate values in the MtF group. Organizational and activational hormone effects on the InM are discussed. PMID- 21618225 TI - Morphological and functional characterization of cholinergic interneurons in the dorsal horn of the mouse spinal cord. AB - Endogenous acetylcholine is an important modulator of sensory processing, especially at the spinal level, where nociceptive (pain-related) stimuli enter the central nervous system and are integrated before being relayed to the brain. To decipher the organization of the local cholinergic circuitry in the spinal dorsal horn, we used transgenic mice expressing enchanced green fluorescent protein specifically in cholinergic neurons (ChAT::EGFP) and characterized the morphology, neurochemistry, and firing properties of the sparse population of cholinergic interneurons in this area. Three-dimensional reconstruction of lamina III ChAT::EGFP neurons based either on their intrinsic fluorescence or on intracellular labeling in live tissue demonstrated that these neurons have long and thin processes that grow preferentially in the dorsal direction. Their dendrites and axon are highly elongated in the rostrocaudal direction, beyond the limits of a single spinal segment. These unique morphological features suggest that dorsal horn cholinergic interneurons are the main contributors to the plexus of cholinergic processes located in lamina IIi, just dorsal to their cell bodies. In addition, immunostainings demonstrated that dorsal horn cholinergic interneurons in the mouse are gamma-aminobutyric acidergic and express nitric oxide synthase, as in rats. Finally, electrophysiological recordings from these neurons in spinal cord slices demonstrate that two-thirds of them have a repetitive spiking pattern with frequent rebound spikes following hyperpolarization. Altogether our results indicate that, although they are rare, the morphological and functional features of cholinergic neurons enable them to collect segmental information in superficial layers of the dorsal horn and to modulate it over several segments. PMID- 21618224 TI - Genetic tracing of Nav1.8-expressing vagal afferents in the mouse. AB - Nav1.8 is a tetrodotoxin-resistant sodium channel present in large subsets of peripheral sensory neurons, including both spinal and vagal afferents. In spinal afferents, Nav1.8 plays a key role in signaling different types of pain. Little is known, however, about the exact identity and role of Nav1.8-expressing vagal neurons. Here we generated mice with restricted expression of tdTomato fluorescent protein in all Nav1.8-expressing afferent neurons. As a result, intense fluorescence was visible in the cell bodies, central relays, and sensory endings of these neurons, revealing the full extent of their innervation sites in thoracic and abdominal viscera. For instance, vagal and spinal Nav1.8-expressing endings were seen clearly within the gastrointestinal mucosa and myenteric plexus, respectively. In the gastrointestinal muscle wall, labeled endings included a small subset of vagal tension receptors but not any stretch receptors. We also examined the detailed innervation of key metabolic tissues such as liver and pancreas and evaluated the anatomical relationship of Nav1.8-expressing vagal afferents with select enteroendocrine cells (i.e., ghrelin, glucagon, GLP-1). Specifically, our data revealed the presence of Nav1.8-expressing vagal afferents in several metabolic tissues and varying degrees of proximity between Nav1.8 expressing mucosal afferents and enteroendocrine cells, including apparent neuroendocrine apposition. In summary, this study demonstrates the power and versatility of the Cre-LoxP technology to trace identified visceral afferents, and our data suggest a previously unrecognized role for Nav1.8-expressing vagal neurons in gastrointestinal functions. PMID- 21618226 TI - General visceral and gustatory connections of the posterior thalamic nucleus of goldfish. AB - Fiber connections of the posterior thalamic nucleus were studied in goldfish. Tracer injections into the rostral part of posterior thalamic nucleus labeled somata and terminals in the medial subnucleus of commissural nucleus of Cajal, secondary general visceral nucleus, a nucleus tentatively identified as the preglomerular general visceral nucleus, torus lateralis, inferior lobar nuclei, preoptic area, dorsal region of medial part of dorsal telencephalic area, and supracommissural part of ventral telencephalic area. Labeled terminals were observed in the lateral subnucleus of commissural nucleus of Cajal; primary gustatory centers, in particular the vagal lobe; and lateral valvular nucleus. Other occasional connections were also observed in a number of structures. The results of tracer injections to the brainstem general visceral and gustatory structures suggested reciprocal connections with the caudal part of posterior thalamic nucleus. These findings suggest that the posterior thalamic nucleus is primarily a general visceral/gustatory structure, serving as a forebrain integration center of visceral information. Descending fibers to the vagal lobe presumably represent a major channel through which the forebrain regulates pharyngeal feeding behavior. PMID- 21618228 TI - Involvement of Galpha(olf)-expressing neurons in the vomeronasal system of Bufo japonicus. AB - Most terrestrial vertebrates possess anatomically distinct olfactory organs: the olfactory epithelium (OE) and the vomeronasal organ (VNO). In rodents, olfactory receptors coupled to Galpha(olf) are expressed in the OE, whereas vomeronasal receptors type 1 (V1R) and vomeronasal receptors type 2 (V2R), coupled to Galpha(i2) and Galpha(o) , respectively, are expressed in the VNO. These receptors and G proteins are thought to play important roles in olfactory perception. However, we previously reported that only V2R and Galpha(o) expression is detected in the Xenopus laevis VNO. As X. laevis spends its entire life in water, we considered that expression of limited types of chemosensory machinery in the VNO might be due to adaptation of the VNO to aquatic life. Thus, we analyzed the expression of G proteins in the VNO and the accessory olfactory bulb (AOB) of the adult Japanese toad, Bufo japonicus, because this species is well adapted to a terrestrial life. By using immunohistochemical analysis in combination with in situ hybridization and DiI labeling, we found that B. japonicus Galpha(olf) and Galpha(o) were expressed in the apical and middle-to basal layer of the vomeronasal neuroepithelium, and that the axons of these Galpha(olf) - and Galpha(o) -expressing vomeronasal neurons projected to the rostral and caudal accessory olfactory bulb, respectively. These results strongly suggest that both the Galpha(olf) - and Galpha(o) -mediated signal transduction pathways function in the B. japonicus VNO. The expression of Galpha(olf) in the B. japonicus VNO may correlate with the detection of airborne chemical cues and with a terrestrial life. PMID- 21618229 TI - Efferent connections of nucleus accumbens subdivisions of the domestic chicken (Gallus domesticus): an anterograde pathway tracing study. AB - Envisaged as a limbic-motor interface, the mammalian nucleus accumbens (Ac) is responsible for motivation, emotionality, and reward mechanisms. As in mammals, Ac of the domestic chick has three subdivisions: the rostral pole (AcR) lying in the rostral part of basal telencephalon, the core (AcC), corresponding to the ventromedial medial striatum, and the shell (AcS), lying ventrally and ventrolaterally to the AcC. Less well known is the connectivity of subdivisions. Here we report on the efferents of Ac subregions, using biotinylated dextran amine as anterograde tracer, deposited into the AcR, AcS, and AcC. The projections of the accumbens subregions mainly overlap in the telencephalon and the diencephalon but differ in the brainstem. In the telencephalon, the main projection sites are the ventral pallidum, the basal nucleus (Meynert), and the nucleus of the diagonal band. The lateral hypothalamus and lateral preoptic area receive strong projections from the AcR and AcS, and weaker projections from the AcC. The AcR and AcC massively innervate the subthalamic nucleus. In the brainstem the bulk of accumbens fibers were found in the compact part of the substantia nigra. All subregions project to the parabrachial region, reticular formation, periaqueductal gray, and the raphe nuclei, with some differences in the weights and subregional distributions. AcR and AcS project extensively to the ventral tegmental area, while AcC sends massive innervation to the solitary and vagal motor nuclei. Overall, the results seem to support the previously suggested distribution of Ac subregions, emphasizing similarities and differences with mammals. PMID- 21618227 TI - Inputs to the midbrain dopaminergic complex in the rat, with emphasis on extended amygdala-recipient sectors. AB - The midbrain dopaminergic neuronal groups A8, A9, A10, and A10dc occupy, respectively, the retrorubral field (RRF), substantia nigra compacta (SNc), ventral tegmental area (VTA), and ventrolateral periaqueductal gray (PAGvl). Collectively, these structures give rise to a mixed dopaminergic and nondopaminergic projection system that essentially permits adaptive behavior. However, knowledge is incomplete regarding how the afferents of these structures are organized. Although the VTA is known to receive numerous afferents from cortex, basal forebrain, and brainstem and the SNc is widely perceived as receiving inputs mainly from the striatum, the afferents of the RRF and PAGvl have yet to be assessed comprehensively. This study was performed to provide an account of those connections and to seek a better understanding of how afferents might contribute to the functional interrelatedness of the VTA, SNc, RRF, and PAGvl. Ventral midbrain structures received injections of retrograde tracer, and the resulting retrogradely labeled structures were targeted with injections of anterogradely transported Phaseolus vulgaris leucoagglutinin. Whereas all injections of retrograde tracer into the VTA, SNc, RRF, or PAGvl produced labeling in many structures extending from the cortex to caudal brainstem, pronounced labeling of structures making up the central division of the extended amygdala occurred following injections that involved the RRF and PAGvl. The anterograde tracing supported this finding, and the combination of retrograde and anterograde labeling data also confirmed reports from other groups indicating that the SNc receives robust input from many of the same structures that innervate the VTA, RRF, and PAGvl. PMID- 21618230 TI - Assembly properties of lamprey neurofilament subunits and their expression after spinal cord transection. AB - In mammals neurofilaments (NF) are formed by coassembly of three subunits: NFL, NFM, and NFH (light, medium, and heavy). It had been believed that lampreys have only one subunit, NF180. However, a previous study showed that NF180 could not self-assemble but could coassemble with rat NFL, suggesting the existence of additional NF subunits in lamprey. More recently, we cloned three additional NF subunits. These new subunits and NF180 have now been transfected in combinations into SW13cl.2Vim(-) cells, which lack endogenous cytoplasmic intermediate filaments. None of the subunits could self-assemble. No combination of NF subunits could form filaments in the absence of lamprey NFL (L-NFL). Assembly occurred at 28 degrees C, but not at 37 degrees C. L-NFL could form thick NF bundles with NF180 but not with NF132 and NF95, which formed only fine filamentous arrays. To determine which parts of the NF subunits are required for filament or bundle formation, we constructed deletion mutants of NF180 and cotransfected them with L-NFL. As with mammalian NF, only constructs with intact head and core domains could form filaments with L-NFL. However, the full length of NF180 was required to form NF bundles. As with NF180, in situ hybridization indicated that mRNA for L-NFL and NF132 was downregulated in identified reticulospinal neurons by 5 weeks after spinal cord transection, but was reexpressed at 10 weeks selectively in those neurons whose axons have a high probability of regenerating. This is consistent with a possible role of NFs in the mechanism of axon regeneration. PMID- 21618231 TI - Connections of the terminal nerve and the olfactory system in two galeomorph sharks: an experimental study using a carbocyanine dye. AB - In elasmobranchs the terminal nerve courses separately from the olfactory nerve. This characteristic makes elasmobranchs excellent models to study the anatomy and function of these two systems. Here we study the neural connections of the terminal nerve and olfactory system in two sharks by experimental tracing methods using carbocyanine dyes. The main projections from the terminal nerve system (consisting of three ganglia in Scyliorhinus canicula) course ipsilaterally to the medial septal nucleus and bilaterally to the ventromedial telencephalic pallial region. Minor terminal nerve projections were also traced ipsilaterally to diencephalic and mesencephalic levels. With regard to the olfactory connections, our results show that in sharks, unlike ray-finned fishes, the primary olfactory projections are mainly restricted to the olfactory bulb. We also performed tracer application to the olfactory bulb in order to analyze the possible central neuroanatomical relationship between the projections of the terminal nerve and the olfactory bulb. In these experiments labeled neurons and fibers were observed from telencephalic to caudal mesencephalic regions. However, we observe almost no overlap between the two systems at central levels. The afferent and the putatively efferent connections of the dogfish olfactory bulb are compared with those previously reported in other elasmobranchs. The significance of the extratelencephalic secondary olfactory projections is also discussed in a comparative context. PMID- 21618232 TI - Segregation of feedforward and feedback projections in mouse visual cortex. AB - Hierarchical organization is a common feature of mammalian neocortex. Neurons that send their axons from lower to higher areas of the hierarchy are referred to as "feedforward" (FF) neurons, whereas those projecting in the opposite direction are called "feedback" (FB) neurons. Anatomical, functional, and theoretical studies suggest that these different classes of projections play fundamentally different roles in perception. In primates, laminar differences in projection patterns often distinguish the two projection streams. In rodents, however, these differences are less clear, despite an established hierarchy of visual areas. Thus the rodent provides a strong test of the hypothesis that FF and FB neurons form distinct populations. We tested this hypothesis by injecting retrograde tracers into two different hierarchical levels of mouse visual cortex (area 17 and anterolateral area [AL]) and then determining the relative proportions of double-labeled FF and FB neurons in an area intermediate to them (lateromedial area [LM]). Despite finding singly labeled neurons densely intermingled with no laminar segregation, we found few double-labeled neurons (~5% of each singly labeled population). We also examined the development of FF and FB connections. FF connections were present at the earliest timepoint we examined (postnatal day 2, P2), while FB connections were not detectable until P11. Our findings indicate that, even in cortices without laminar segregation of FF and FB neurons, the two projection systems are largely distinct at the neuronal level and also differ with respect to the timing of their axonal outgrowth. PMID- 21618233 TI - Motoneurons, DUM cells, and sensory neurons in an insect thoracic ganglion: a tracing study in the stick insect Carausius morosus. AB - Anatomical features of leg motoneurons, dorsal unpaired median (DUM) cells, and sensory neurons in stick insect mesothoracic ganglia were examined using fluorescent dye backfills of lateral nerves. Structures were analyzed in whole mounts of ganglia and transverse sections. Numbers of motoneurons and details of their structure by far exceed previously published data. The general neuroanatomical layout of motoneurons matches the general orthopteran pattern. Cell bodies of excitatory motoneurons form clusters in the lateral cortex, dendrites branch mainly in the dorsal neuropil. We identified nine DUM cells, six of which have axons in nerve nl5. Most sensory fibers terminate in the ventral association center (VAC). Twenty-three small cell bodies located close to the soma of the fast extensor tibiae motoneuron likely belong to strand receptors. Labeled structures are compared with previously published data from stick insects and other orthopterous insects. PMID- 21618234 TI - Stereological analysis of the rat and monkey amygdala. AB - The amygdala is part of a neural network that contributes to the regulation of emotional behaviors. Rodents, especially rats, are used extensively as model organisms to decipher the functions of specific amygdala nuclei, in particular in relation to fear and emotional learning. Analysis of the role of the nonhuman primate amygdala in these functions has lagged work in the rodent but provides evidence for conservation of basic functions across species. Here we provide quantitative information regarding the morphological characteristics of the main amygdala nuclei in rats and monkeys, including neuron and glial cell numbers, neuronal soma size, and individual nuclei volumes. The volumes of the lateral, basal, and accessory basal nuclei were, respectively, 32, 39, and 39 times larger in monkeys than in rats. In contrast, the central and medial nuclei were only 8 and 4 times larger in monkeys than in rats. The numbers of neurons in the lateral, basal, and accessory basal nuclei were 14, 11, and 16 times greater in monkeys than in rats, whereas the numbers of neurons in the central and medial nuclei were only 2.3 and 1.5 times greater in monkeys than in rats. Neuron density was between 2.4 and 3.7 times lower in monkeys than in rats, whereas glial density was only between 1.1 and 1.7 times lower in monkeys than in rats. We compare our data in rats and monkeys with those previously published in humans and discuss the theoretical and functional implications that derive from our quantitative structural findings. PMID- 21618235 TI - Two distinct types of ON directionally selective ganglion cells in the rabbit retina. AB - Mammalian retinas contain about 20 types of ganglion cells that respond to different aspects of the visual scene, including the direction of motion of objects in the visual field. The rabbit retina has long been thought to contain two distinct types of directionally selective (DS) ganglion cell: a bistratified ON-OFF DS ganglion cell that responds to onset and termination of light, and an ON DS ganglion cell, which stratifies only in the ON layer and responds only to light onset. This division is challenged by targeted recordings from rabbit retina, which indicate that ON DS ganglion cells occur in two discriminably different types. One of these is strongly tracer-coupled to amacrine cells; the other is never tracer-coupled. These two types also differ in branching pattern, stratification depth, relative latency, and transience of spiking. The sustained, uncoupled ON DS cell ramifies completely within the lower cholinergic band and responds to nicotine with continuous firing. In contrast, the transient, coupled ON DS ganglion cell stratifies above the cholinergic band and is not positioned to receive major input from cholinergic amacrine cells, consistent with its modest response to the cholinergic agonist nicotine. Much data have accrued that directional responses in the mammalian retina originate via gamma-aminobutyric acid (GABA) release from the dendrites of starburst amacrine cells (Euler et al., 2002). If there is an ON DS ganglion cell that does not stratify in the starburst band, this suggests that its GABA-dependent directional signals may be generated by a mechanism independent of starburst amacrine cells. PMID- 21618236 TI - Neurochemical phenotypes of myenteric neurons in the rhesus monkey. AB - Understanding the neurochemical composition of the enteric nervous system (ENS) is critical for elucidating neurological function in the gastrointestinal (GI) tract in health and disease. Despite their status as the closest models of human neurological systems, relatively little is known about enteric neurochemistry in nonhuman primates. We describe neurochemical coding of the enteric nervous system, specifically the myenteric plexus, of the rhesus monkey (Macaca mulatta) by immunohistochemistry and directly compare it to human tissues. There are considerable differences in the myenteric plexus along different segments of the monkey GI tract. While acetylcholine neurons make up the majority of myenteric neurons in the stomach (70%), they are a minority in the rectum (47%). Conversely, only 22% of gastric myenteric neurons express nitric oxide synthase (NOS) compared to 52% in the rectum. Vasoactive intestinal peptide (VIP) is more prominent in the stomach (37%) versus the rest of the GI tract (~10%), and catecholamine neurons are rare (~1%). There is significant coexpression of NOS and VIP in myenteric neurons that is more prominent in the proximal GI tract. Taken as a whole, these data provide insight into the neurochemical anatomy underlying GI motility. While overall similarity to other mammalian species is clear, there are some notable differences between the ENS of rhesus monkeys, humans, and other species that will be important to take into account when evaluating models of human diseases in animals. PMID- 21618237 TI - Monosynaptic inputs to ErbB4-expressing inhibitory neurons in mouse primary somatosensory cortex. AB - Previous reports have described inputs to the somatosensory cortex (S1) in mouse or rat using retrograde or anterograde tracers. Such studies do not, however, reveal which particular cell types within the S1 cortex receive direct monosynaptic connections from these input sources. Here we describe the monosynaptic inputs to a subpopulation of mouse S1 inhibitory neurons that express ErbB4. We used a previously described "bridge protein," composed of the ErbB4 ligand, neuregulin (NRG1), fused to the avian viral receptor TVB (TVB NRG1), along with EnvB pseudotyped lentivirus (LV) and rabies virus (RV), to selectively coinfect ErbB4-expressing neurons (Choi et al. [2010] Proc Natl Acad Sci U S A 107:16703-16708). The RV had its glycoprotein gene deleted and replaced with mCherry, so that infected cells express mCherry and the virus cannot spread without provision of rabies glycoprotein (RG) by transcomplementation. The LV encoded and expressed RG to allow transcomplementation in coinfected neurons, so that the RV could spread transsynaptically and label their direct monosynaptic inputs. The RV could not spread beyond the direct inputs, due to the lack of RG in presynaptic cells. This method revealed long-range connections from thalamus, nucleus basalis, raphe, and distant cortical areas, including ipsilateral motor, secondary somatosensory, retrosplenial, and perirhinal cortex and contralateral S1. In addition, local connections from ipsilateral pyramidal neurons within S1 were labeled. These input sources account for all of the known inputs to S1 described with standard tracers, suggesting that the subpopulation of ErbB4 positive inhibitory neurons infected using the TVB-NRG1 bridge protein receives inputs indiscriminately from S1 input sources. PMID- 21618238 TI - Subcellular localization of Patched and Smoothened, the receptors for Sonic hedgehog signaling, in the hippocampal neuron. AB - Cumulative evidence suggests that, aside from patterning the embryonic neural tube, Sonic hedgehog (Shh) signaling plays important roles in the mature nervous system. In this study, we investigate the expression and localization of the Shh signaling receptors, Patched (Ptch) and Smoothened (Smo), in the hippocampal neurons of young and mature rats. Reverse transcriptase-polymerase chain reaction and immunoblotting analyses show that the expression of Ptch and Smo remains at a moderate level in young postnatal and adult brains. By using immunofluorescence light microscopy and immunoelectron microscopy, we examine the spatial distribution of Ptch and Smo within the hippocampal neurons. In young developing neurons, Ptch and Smo are present in the processes and are clustered at their growth cones. In mature neurons, Ptch and Smo are concentrated in dendrites, spines, and postsynaptic sites. Synaptic Ptch and Smo often co-exist with unusual structures-synaptic spinules and autophagosomes. Our results reveal the anatomical organization of the Shh receptors within both the young and the mature hippocampal neurons. PMID- 21618240 TI - The roles of fascins in health and disease. AB - Fascins are actin-binding proteins that cross-link filamentous actin into tightly packed parallel bundles. These bundles are important for the organization and morphology of an extremely diverse set of sub-cellular structures that include dynamic and stable cell-surface protrusions, stress fibres, and the specialized actin bundles of photoreceptor and stereocilia cells. In this review, we discuss the fascin gene family and its evolution, the actin-bundling activity of fascins and the molecular pathways by which it is regulated, and the role of the diverse actin/fascin structures in normal cellular processes. We discuss the mechanisms by which fascins contribute to disease pathologies, especially cancer, where fascin-1 is emerging as a novel therapeutic target in carcinoma metastasis. PMID- 21618239 TI - Thalamocortical projections to rat auditory cortex from the ventral and dorsal divisions of the medial geniculate nucleus. AB - The ventral and dorsal medial geniculate (MGV and MGD) constitute the major auditory thalamic subdivisions providing thalamocortical inputs to layer IV and lower layer III of auditory cortex. No quantitative evaluation of this projection is available. Using biotinylated dextran amine (BDA)/biocytin injections, we describe the cortical projection patterns of MGV and MGD cells. In primary auditory cortex the bulk of MGV axon terminals are in layer IV/lower layer III with minor projections to supragranular layers and intermediate levels in infragranular layers. MGD axons project to cortical regions designated posterodorsal (PD) and ventral (VA) showing laminar terminal distributions that are quantitatively similar to the MGV-to-primary cortex terminal distribution. At the electron microscopic level MGV and MGD terminals are non-gamma-aminobutyric acid (GABA)ergic with MGD terminals in PD and VA slightly but significantly larger than MGV terminals in primary cortex. MGV/MGD terminals synapse primarily onto non-GABAergic spines/dendrites. A small number synapse on GABAergic structures, contacting large dendrites or cell bodies primarily in the major thalamocortical recipient layers. For MGV projections to primary cortex or MGD projections to PD or VA, the non-GABAergic postsynaptic structures at each site were the same size regardless of whether they were in supragranular, granular, or infragranular layers. However, the population of MGD terminal-recipient structures in VA were significantly larger than the MGD terminal-recipient structures in PD or the MGV terminal-recipient structures in primary cortex. Thus, if terminal and postsynaptic structure size indicate strength of excitation then MGD to VA inputs are strongest, MGD to PD intermediate, and MGV to primary cortex the weakest. PMID- 21618241 TI - Free fatty acid receptors FFAR1 and GPR120 as novel therapeutic targets for metabolic disorders. AB - Free fatty acids (FFAs) are not only essential nutritional components, but they also act as signaling molecules in various physiological processes. Recently, a G protein-coupled receptor deorphanizing strategy has successfully identified a family of receptors that are activated by FFAs. FFA receptors (FFARs) are proposed to play critical roles in a variety of physiological and pathophysiological processes, especially in metabolic disorders. Among the FFARs, FFAR1 (GPR40) and GPR120 are activated by medium- and long-chain FFAs. FFAR1 facilitates glucose-stimulated insulin secretion from pancreatic beta-cells, whereas GPR120 regulates the secretion of glucagon-like peptide-1 in the intestine, as well as insulin sensitivity in macrophages. Because these receptors are potential therapeutic targets for metabolic disorders such as type 2 diabetes, selective ligands have been developed. In this review, we discuss recent advances in the identification of ligands, structure activity relationships, and pharmacological characterization of FFAR1 and GPR120, and we present a summary of recent progress in understanding their physiological roles and their potential as drug targets. PMID- 21618242 TI - Standardized pretreatment breast MRI--accuracy and influence on mastectomy decisions. AB - BACKGROUND AND OBJECTIVES: Routine pretreatment breast magnetic resonance imaging in newly diagnosed cancer patients remains controversial. We assess MRI accuracy and influence on mastectomy decisions after institution of standardized pretreatment MRI. METHODS: A prospectively collected database of 74 consecutive new invasive breast cancer patients with pretreatment breast MRI was reviewed for treatment choice, radiologic, and pathologic results. Thirty-eight of 72 patients with available surgical records underwent mastectomy. Mastectomy preoperative and operative electronic records were reviewed for treatment decision analysis. RESULTS: Seventeen of 72 (23.6%) invasive breast cancer patients were likely influenced to undergo mastectomy by new information from MRI. MRI reported that the multifocal/multicentric (MF/MC) rate was 20 of 72 (27.8%) versus 19 of 72 (26.4%) by surgical pathology. MRI sensitivity for MF/MC disease was 89.5% versus 11.8% for mammography. MRI specificity was 84.2%. All three false positives declined recommended preoperative biopsies. MRI MF/MC diagnosis highly correlated with pathology results, P < 0.001. CONCLUSIONS: Increased mastectomy rate from 29 to 52.8% after standardization of pre-treatment breast MRI for invasive cancer is largely due to MRI findings of increased extent of disease. These MRI findings correlate well with pathologic findings and appear to justify the performance of mastectomies in these patients. PMID- 21618243 TI - Use of the "Secrea (HogyTM)" sponge spacer in thoracoscopic surgery for lung cancer. PMID- 21618244 TI - Regional chemotherapy for pancreatic cancer. PMID- 21618245 TI - Does surgery or radiation therapy impact survival for patients with extrapulmonary small cell cancers? AB - BACKGROUND AND OBJECTIVES: Extrapulmonary small cell carcinomas (EPSCC) are rare tumors where therapy remains poorly defined. We sought to determine the impact of surgical extirpation and radiation therapy for outcomes of EPSCC. METHODS: The Surveillance, Epidemiology, and End Results (SEER) database was queried for patients with EPSCC which were further categorized by site and evaluated for survival by specific treatment strategy. RESULTS: We identified 94,173 patients with small cell carcinoma of which 88,605 (94.1%) and 5,568 (5.9%) had pulmonary small cell carcinoma and EPSCC, respectively. EPSCC patients were subdivided by site with the following proportions: genitourinary (24.1%), gastrointestinal (22.1%), head and neck (7.1%), breast (4%), and miscellaneous (42.7%). Overall EPSSC and specifically gastrointestinal disease had significantly improved median, 5- and 10-year survival with surgery and/or radiation for all stages and sizes. For all EPSCCs multivariate analysis revealed age (>50), gender (female), stage (regional, distant), radiation, and surgery to be independent predictors of survival. CONCLUSIONS: Although outcomes for EPSCC remains poor, both surgery and radiation is shown to significantly improve median, 5- and 10-year survival rates. EPSCC patients who are potential candidates for surgical resection or radiation therapy may benefit from these treatments. PMID- 21618246 TI - Identification and validation of Kallikrein-ralated peptidase 11 as a novel prognostic marker of gastric cancer based on immunohistochemistry. AB - BACKGROUND AND OBJECTIVES: It is important to identify and validate the differentially expressed genes in gastric cancer to screen diagnostic and/or prognostic tumor markers. METHODS: cDNA expression microarray, gene set enrichment analysis, and bioinformatics approaches were used to screen the differentially expressed genes between gastric cancer tissues and adjacent non cancerous mucosa. A novel candidate prognostic marker, Kallikrein-related peptidase 11 (KLK11), was validated in 400 Chinese gastric cancer patients. KLK11 expression in gastric cancer tissues was detected using real-time PCR and Western blot. KLK11 protein expression was further analyzed by immunostaining on tissue microarray, followed with clinicopathological significance and survival analysis. RESULTS: KLK11 expression was significantly decreased in gastric cancer compared with that in normal gastric mucosa (P<0.001). Furthermore, KLK11 expression was much lower in poorly differentiated cancer samples than that in well differentiated group (P<0.01). Survival analysis showed that negative KLK11 expression was associated with nearly fivefold increased risk of distant metastasis after curative gastrectomy (HR 4.65, P<0.01). Multivariate Cox regression analysis showed that KLK11 expression emerged as a significant independent prognostic factor for disease-free survival and overall survival (P<0.05). CONCLUSIONS: The results indicated that KLK11 expression was decreased in gastric cancer and might serve as a novel independent prognostic marker. PMID- 21618247 TI - Long-term outcomes and survival after laparoscopy-assisted distal gastrectomy for gastric cancer: three-year survival analysis of a single-center experience in Korea. AB - BACKGROUND AND OBJECTIVES: Laparoscopy-assisted distal gastrectomy (LADG) has been established as an alternative treatment for early gastric cancer (EGC) because of excellent short-term results. However, only a few reports have considered the long-term outcomes of LADG. In this study, we investigated the 3 year outcome and survival of patients who underwent LADG. METHODS: We assessed 182 patients with gastric adenocarcinoma who underwent LADG. The indication for LADG was confined to EGCs (T1N0 or T1N1 cases). The clinicopathological characteristics and long-term survival data of all patients were analyzed. RESULTS: The overall morbidity and mortality rates of the patients were 11% and 0%, respectively. An analysis of the final pathological stages of the patients revealed that 160 had stage Ia, 20 had stage Ib, and only 2 had stage II. The median follow-up period was 44 months (range, 2-73 months), and there were two recurrences. Five patients died of other causes, but no patients died of a gastric cancer recurrence. The 3-year overall and disease-specific survival rates were 97.3% and 100%, respectively. CONCLUSIONS: LADG for EGC is acceptable in terms of both short- and long-term outcomes. Thus, LADG can be considered a primary treatment for EGC. PMID- 21618248 TI - Surgical outcomes for gastric cancer patients with intraperitoneal free cancer cell, but no macroscopic peritoneal metastasis. AB - BACKGROUND AND OBJECTIVES: Gastric cancer patients with intraperitoneal cancer cells have extremely poor prognoses, because they frequently develop peritoneal metastasis. The aim of the current study is to clarify the clinicopathologic characteristics of patients with intraperitoneal free cancer cells who do not have recurrences. METHODS: This study examined 1,985 gastric adenocarcinoma patients who underwent gastrectomies at our institution between January 1975 and December 2000. RESULTS: The 5-year survival rate of advanced gastric cancer patients with intraperitoneal free cancer cells, but no macroscopic peritoneal metastases (P0Cy1), is 15.3%. Multivariate analysis identified only lymph node metastasis as an independent prognostic indicator in P0Cy1 patients. The 5-year survival rates of P0Cy1 patients were 62.5%, 19.1%, 17.6%, and 9.4% in n0, n1, n2, and n3 patients, respectively. Peritoneal recurrence was observed significantly more often in P0Cy1 patients than in advanced gastric cancer patients without intraperitoneal free cancer cells. CONCLUSIONS: Our data indicate that prognoses of P0Cy1 patients without lymph node metastasis are good, whereas the prognoses of those with lymph node metastasis are extremely poor. Therefore, radical surgery and adequate adjuvant chemotherapy should be performed for P0Cy1 patients with no lymph node metastasis. PMID- 21618249 TI - Expression and prognostic significance of CEACAM6, ITGB1, and CYR61 in peripheral blood of patients with gastric cancer. AB - BACKGROUND: We examined CEACAM6, ITGB1, and cyr61 concentrations from patients with gastric cancers (GCs) to assess their clinical application for diagnosing and monitoring diseases. METHODS: Real-time reverse transcription-polymerase chain reaction was used to detect the expressions of CEA, CEACAM6, ITGB1, IGF1R, CK20, cyr61, and S100A4 in peripheral blood karyocyte from 82 patients with GC, 24 patients with recurrence, and 37 healthy volunteers. Receiver operating characteristics (ROC) curves were constructed. RESULTS: There were significant association between these CEACAM6, ITGB1, and cyr61 and TNM Stages and distant metastasis. The AUC of CEACAM6 was 0.884 +/- 0.044 (P = 0.0001), the AUC of cyr61 was 0.833 +/- 0.047 (P = 0.0001), and the AUC of ITGB1 was 0.838 +/- 0.042 (P = 0.0001) by differentiating preoperative GC patients from healthy volunteers from ROC curve analysis. The AUC of CEACAM6 was 0.761 +/- 0.066 (P = 0.001), the AUC of CYR61 was 0.762 +/- 0.063 (P = 0.001), and the AUC of ITGB1 was 0.824 +/- 0.051 (P = 0.0001), by differentiating recurrence of GC from healthy volunteers from ROC curve analysis. CONCLUSION: The method of detecting the expression of CEACAM6, ITGB1, and CYR61 in peripheral blood of GC patients was more sensitive than CEA, IGF1R, CK20, and S100A4 for the early diagnosis of metastasis and recurrence. PMID- 21618250 TI - The peak-standardized uptake value (P-SUV) by preoperative positron emission tomography-computed tomography (PET-CT) is a useful indicator of lymph node metastasis in gastric cancer. AB - BACKGROUND AND OBJECTIVES: Little data is currently available on the usefulness of peak-standardized uptake value (P-SUV) by positron emission tomography computed tomography (PET-CT) in gastric cancer. The purpose of the present study was to evaluate the value of PET-CT for the preoperative evaluation of patients with gastric cancer. The aim of this study was to assess the relation of between primary tumor P-SUV, as determined by preoperative PET-CT, and lymph node metastasis in gastric cancer. METHODS: From December 2007 to March 2010, we analyzed the PET-CT of 147 patients that underwent gastrectomy for gastric cancer. P-SUV in PET-CT were measured by single nuclear medicine physician. Statistical analysis was performed to determine relations between clinicopathologic parameters including P-SUV and lymph node metastasis using the chi-square test, the independent t-test, and using logistic regression analysis. RESULTS: Age, tumor depth, tumor size, and lymph node metastasis were found to be associated with primary tumor P-SUV by PET-CT (P=0.009, <0.001, <0.001, and <0.001, respectively). No association was found between P-SUV and tumor histology or tumor location (P=0.099). Advanced gastric cancer was found to have a higher P SUV than early gastric cancer, and a higher P-SUV was found to be associated with lymph node metastases by both univariate and multivariate analysis. CONCLUSIONS: P-SUV of primary tumor could be an independent indicator of lymph node metastasis in gastric cancer. Gastric surgeons should pay more attention to the dissection of lymph nodes when primary tumors have higher P-SUV values by PET-CT. PMID- 21618251 TI - Elevated preoperative neutrophil to lymphocyte ratio predicts poor survival following resection in late stage gastric cancer. AB - BACKGROUND: Elevated neutrophil to lymphocyte ratio (N/L ratio) has been shown to be a prognostic indicator in various cancers. We aimed to investigate the prognostic significance of the preoperative N/L ratio in late stage gastric cancer. METHODS: From April 2004 to August 2007, 293 patients who had undergone gastrectomy with curative intent for the AJCC/UICC TNM Stage III or IV gastric cancer were included. N/L ratio was calculated from lymphocyte and neutrophil counts on routine blood tests taken prior to surgery. RESULTS: The median follow up time for surviving patients was 38.2 months (4.2-65.5 months) and median preoperative N/L ratio was 2.06 (range 0.47-19.73). Subjects were dichotomized at the N/L value of 2.0. A multivariate analysis established a significant relationship between the N/L ratio and overall survival (HR=1.609; 95% confidence interval, CI, 1.144-2.264; P=0.006). The cutoff value up to 3.0, the value of 75 percentiles, showed a significant prognostic effect on disease-free survival (HR=1.654; 95% CI, 1.088-2.515; P=0.019). CONCLUSIONS: The results suggest that the elevated preoperative N/L ratio predicts poor disease-free and overall survival following resection for late stage gastric cancer. It may be utilized as a simple, reliable prognostic factor for risk stratification and will provide better treatment allocation. PMID- 21618252 TI - New real-time loop recorder diagnosis of symptomatic arrhythmia via telemedicine. AB - BACKGROUND: One disadvantage of current loop recorders is the long interval between recording an electrocardiogram (ECG), establishing a diagnosis, and taking appropriate medical measures. The Cardio R loop recorder transmits cardiac recordings by cellular communication at the push of a button. Users can concomitantly relay symptoms, thereby providing a symptom/cardio-rhythm correlation. HYPOTHESIS: The Cardio R is capable of early detection of cardio electrical events that could account for patients' symptoms. METHODS: This observational study was designed to evaluate patients who were referred from community physicians/cardiologists for evaluation of various cardiac symptoms that were not observed by regular office ECGs or traditional 24-hour Holter cardiac monitoring. Transmitted recordings were instantly displayed on a monitor for immediate diagnosis by the on-duty medical team at SHL-Telemedicine's call center. Abnormal tracings, especially when accompanied by symptoms selected from the prepared list, enabled the staff to instruct the subscriber, notify their physician, and/or dispatch a mobile intensive care unit to the scene. RESULTS: Between January 2009 and August 2010, there were 17 622 ECG transmissions received from 604 patients (age range, 10-95 years) who completed a 1-month trial with the Cardio R device. Palpitation, presyncope, and chest pain were the leading complaints. A disturbance in rhythm that could account for symptoms occurred during recording in 49% cases and was displayed within 7 minutes in 93% of them. No longer than 2 days elapsed from recording onset to diagnosis. CONCLUSIONS: The Cardio R device enables prompt ECG confirmation/exclusion of a probable arrhythmic cause of symptoms, enabling rapid intervention for cardiac relevant complaints. PMID- 21618253 TI - Three-residue loop closure in proteins: a new kinematic method reveals a locus of connected loop conformations. AB - The closure of a three-residue loop was studied using a developed kinematic method. It was shown that there are infinite number of three-residue loops (a locus of conformations), which can connect two segments of a polypeptide. This adds to the current understanding of a finite number of conformations for three residue loop-closure. In the developed method, some of the equations can be solved analytically to reduce the computation cost. Benefiting from the reduced computation time, we determined all the relative positions of two polypeptide segments that can be connected by a three-residue loop. PMID- 21618254 TI - Prenatal interventions for fetal lung lesions. AB - The widespread availability of high resolution ultrasound equipment and almost universal routine anatomy scanning in all pregnant women in the developed world has lead to increased detection of abnormalities in the fetal thorax. Already in the 1980s, large pleural effusions and significant macrocystic lesions in the fetus were easily detected on ultrasound. However, smaller lung tumours were often missed. Nowadays, fetal medicine centres receive many referrals for evaluation of fetal lung lesions, of which the most common are congenital cystic adenomatoid malformation and bronchopulmonary sequestration. Almost invariably, both the parents and the referring physicians experience anxiety after detection of large lung masses in the fetus. However, the vast majority of the currently detected fetal lung lesions have an excellent prognosis without the need for prenatal intervention. In the small group of fetuses in which the prognosis is poor, almost exclusively those with concomitant fetal hydrops and cardiac failure, several options for fetal therapy exist, often with a more than 50% survival rate. Indications, techniques, complications and outcomes of these interventions will be described in this review. PMID- 21618255 TI - Organ targeted prenatal gene therapy--how far are we? AB - Prenatal gene therapy aims to deliver genes to cells and tissues early in prenatal life, allowing correction of a genetic defect, before long-term tissue damage has occurred. In contrast to postnatal gene therapy, prenatal application can target genes to a large population of dividing stem cells, and the smaller fetal size allows a higher vector-to-target cell ratio to be achieved. Early gestation delivery may allow the development of immune tolerance to the transgenic protein which would facilitate postnatal repeat vector administration if needed. Targeting particular organs will depend on manipulating the vector to achieve selective tropism and on choosing the most appropriate gestational age and injection method for fetal delivery. Intra-amniotic injection reaches the skin, and other organs that are bathed in the fluid however since gene transfer to the lung and gut is usually poor more direct injection methods will be needed. Delivery to the liver and blood can be achieved by systemic delivery via the umbilical vein or peritoneal cavity. Gene transfer to the central nervous system in the fetus is difficult but newer vectors are available that transduce neuronal tissue even after systemic delivery. PMID- 21618258 TI - Experience with local lymph node assay performance standards using standard radioactivity and nonradioactive cell count measurements. AB - The local lymph node assay (LLNA) is the preferred test for identification of skin-sensitizing substances by measuring radioactive thymidine incorporation into the lymph node. To facilitate acceptance of nonradioactive variants, validation authorities have published harmonized minimum performance standards (PS) that the alternative endpoint assay must meet. In the present work, these standards were applied to a variant of the LLNA based on lymph node cell counts (LNCC) run in parallel as a control with the standard LLNA with radioactivity measurements, with threshold concentrations (EC3) being determined for the sensitizers. Of the 22 PS chemicals tested in this study, 21 yielded the same results from standard radioactivity and cell count measurements; only 2-mercaptobenzothiazole was positive by LLNA but negative by LNCC. Of the 16 PS positives, 15 were positive by LLNA and 14 by LNCC; methylmethacrylate was not identified as sensitizer by either of the measurements. Two of the six PS negatives tested negative in our study by both LLNA and LNCC. Of the four PS negatives which were positive in our study, chlorobenzene and methyl salicylate were tested at higher concentrations than the published PS, whereas the corresponding concentrations resulted in consistent negative results. Methylmethacrylate and nickel chloride tested positive within the concentration range used for the published PS. The results indicate cell counts and radioactive measurements are in good accordance within the same LLNA using the 22 PS test substances. Comparisons with the published PS results may, however, require balanced analysis rather than a simple checklist approach. PMID- 21618259 TI - Dark cell change of the cerebellar Purkinje cells induced by terbutaline under transient disruption of the blood-brain barrier in adult rats: morphological evaluation. AB - This study aimed to establish a cerebellar degeneration animal model and to characterize the dark cell change of Purkinje cells. We hypothesized that terbutaline, a beta2-adrenoceptor agonist, induces cerebellar degeneration not only in neonatal rats, but also in adult rats. Nine-week-old adult male Sprague Dawley rats were anesthetized and infused with 25% mannitol via the left common carotid artery. Thirty seconds later, terbutaline was infused via the same artery. Dark-stained Purkinje cells were observed in the entire cerebellum on day 3. Prominent Bergmann glial cells accompanied by swelling of the glial processes were present, and were closely associated with the dark-stained Purkinje cells. These findings were found continuously throughout day 30. Ultrastructurally, dilated Golgi vesicles and/or endoplasmic reticulum and large lamella bodies were present in both severely changed and slightly changed Purkinje cells. Bergmann glial cells in the area of synaptic contacts of the severely changed Purkinje cells showed swelling. The Bergmann glial process in close contact with the slightly changed Purkinje cell dendrite in molecular layer showed slight swelling, and large lamella bodies in the dendrite were observed close to the dendritic spines. These findings may suggest that terbutaline induced a failure of Bergmann glial cell and resulted in dark cell degeneration of the Purkinje cells due to glutamate excitotoxicity. PMID- 21618260 TI - Virological breakthrough and resistance in patients with chronic hepatitis B receiving nucleos(t)ide analogues in clinical practice. AB - Virological breakthrough (VBT) is the first manifestation of antiviral drug resistance during nucleos(t)ide analogue (NUC) treatment of chronic hepatitis B (CHB), but not all VBTs are due to drug resistance. This study sought to determine the incidence of VBT and genotypic resistance (GR) in patients with CHB who were receiving NUCs in clinical practice. Records of patients with CHB who were receiving NUCs were reviewed. All patients with VBT were tested for drug resistance mutations. Of 148 patients included, 73% were men and mean age was 44.9 years. During a mean follow-up of 37.5 +/- 20.1 months, 39 (26%) patients had at least 1 VBT. Of these 39 patients, 15 (38%) were not confirmed to have VBT on retesting, and 10 of these 15 had no evidence of GR. The cumulative probability of VBT, confirmed VBT, and GR at 5 years was 46.1%, 29.7%, and 33.9%, respectively. In multivariate analysis, failure to achieve undetectable hepatitis B virus (HBV) DNA was the only factor significantly associated with VBT. Among the 10 patients who had VBT but no confirmed VBT or GR and who were maintained on the same medications, serum HBV DNA decreased in all 10, and nine had undetectable HBV DNA at a mean of 6.8 months after the VBT. Four patients had persistently undetectable HBV DNA, six had transient increase in HBV DNA during follow-up, and none had GR. CONCLUSION: VBT was common in patients with CHB receiving NUCs in clinical practice, but nearly 40% of the VBTs were not related to antiviral drug resistance. Counseling of patients with CHB on medication adherence and confirmation of VBT and/or GR can avoid unnecessary changes in antiviral medications. PMID- 21618261 TI - Reviving pegylated interferon as a therapeutic agent for hepatitis D: no more room for nucleos(t)ides? AB - BACKGROUND: Chronic infection with hepatitis B virus and hepatitis delta virus (HDV) results in the most severe form of viral hepatitis. There is no currently approved treatment. We investigated the safety and efficacy of 48 weeks of treatment with peginterferon alfa-2a plus adefovir dipivoxil, peginterferon alfa 2a alone,and adefovir dipivoxil alone. METHODS: We conducted a randomized trial in which 31 patients with HDV infection received treatment with 180 lg of peginterferon alfa-2a weekly plus 10 mgof adefovir daily, 29 received 180 lg of peginterferon alfa-2a weekly plus placebo, and 30 received 10 mg of adefovir alone weekly for 48 weeks. Follow-up was conducted for an additional 24 weeks. Efficacy end points included clearance of HDV RNA,normalization of alanine aminotransferase levels, and a decline in levels of hepatitis B surface antigen (HBsAg). RESULTS: The primary endpoint-normalization of alanine aminotransferase levels and clearance of HDV RNA at week 48--was achieved in two patients in the group receiving peginterferon alfa-2a plus adefovir and two patients in the group receiving peginterferon alfa-2a plus placebo but in none of the patients in the group receiving adefovir alone. At week 48, the test for HDV RNA was negative in 23% of patients in the first group, 24% of patients in the second, and none of those in the third (P1/4 0.006 for the comparison of the first and third groups; P1/4 0.004 for the comparison of the second and third). The efficacy of peginterferon alfa-2a was sustained for 24 weeks after treatment, with 28% of the patients receiving peginterferon alfa-2a plus adefovir or peginterferon alfa-2a alone having negative results on HDV-RNA tests; none of the patients receiving adefovir alone had negative results. A decline in HBsAg levels of more than 1 log10 IU per milliliter from baseline to week 48 was observed in 10 patients in the first group, 2 in the second, and none in the third (P<0.001 for the comparison of the first and third groups and P1/40.01 for the comparison of the first and second). CONCLUSIONS: Treatment with peginterferon alfa-2a for 48 weeks, with or without adefovir, resulted in sustained HDV RNA clearance in about one quarter of patients with HDV infection. PMID- 21618262 TI - Genome-wide association studies in primary sclerosing cholangitis: still more questions than answers? AB - Primary sclerosing cholangitis (PSC) is a chronic bile duct disease affecting 2.4 7.5% of individuals with inflammatory bowel disease. We performed a genome-wide association analysis of 2,466,182 SNPs in 715 individuals with PSC and 2962 controls,followed by replication in 1025 PSC cases and 2174 controls. We detected non-HLA associations at rs3197999 in MST1 and rs6720394 near BCL2L11 (combined P = 1.1 x (10(-16) and P = 4.1 x 10(-8) respectively). PMID- 21618263 TI - A woman with improving cholestasis but hepatitis aggravating. PMID- 21618266 TI - Nonalcoholic fatty liver disease activity score and Brunt's pathologic criteria for the diagnosis of nonalcoholic steatohepatitis: what do they mean and do they agree? PMID- 21618267 TI - A hairy business-periostracal hair formation in two species of helicoid snails (gastropoda, stylommatophora, helicoidea). AB - In molluscs, the calcareous shell is covered externally by a thin organic layer, the periostracum. The periostracum of some pulmonate species is of special taxonomic interest because it bears distinct microscale architectures. Where and how these structures are formed is as yet unknown. Using histological sections through their shells, gelatin cuts, and live observations I studied the pattern by which the periostracal hair-like projections in two helicoid land snail species are secreted and evenly arranged on the shell. The results indicate a complex mechanism: a hair is formed in the periostracal groove independently of the periostracum, after which it is attached to the edge of the shell, drawn out of the tissue, and finally swivelled to the upper side of the periostracum. Upon further growth of the periostracum, the hairs are finally fixed upright on the shell. PMID- 21618268 TI - Histology of tooth attachment tissues and plicidentine in Varanus (Reptilia: Squamata), and a discussion of the evolution of amniote tooth attachment. AB - Few recent studies have examined the histological basis for tooth attachment in squamates. In the past few years, a surge of interest in this topic has led to the intriguing suggestion that the major tissues derived from the tooth germ (enamel, dentine, cementum and alveolar bone), are conservative and are present in all amniotes. In this study, we describe the histology and development of the tooth attachment complex in Varanus rudicollis, the rough-neck monitor. We provide the first published evidence for the role of cementum and alveolar bone in tooth attachment in varanoid lizards. In Varanus, cementum is deposited on the external surface of the tooth root as well as at the base of the tooth, where it plays a role in the attachment of the tooth to the jawbone. Alveolar bone is also involved in tooth ankylosis. Our results support the hypothesis that the major tooth germ tissues are found in all amniotes. We provide insights into the structure and development of plicidentine, defined as infolding of the dentine around the tooth base. This feature is unique to varanoids among extant tetrapods and is the third tissue implicated in tooth attachment in Varanus. Plicidentine develops asymmetrically along the labial-lingual axis of a tooth. Varanus is characterized by the presence of both primary and higher-order lamellae, which anastomose to form a honeycomb-like surface that then interacts with the more basal attachment tissues. PMID- 21618269 TI - Functional anatomy of scent glands in Paranemastoma quadripunctatum (Opiliones, Dyspnoi, Nemastomatidae). AB - The morphological organization and functional anatomy of prosomal defensive (scent) glands in Paranemastoma quadripunctatum, a representative of the dyspnoid harvestmen, was investigated by means of histological semithin sections, software based 3D-reconstruction and scanning electron microscopy. Scent glands comprise large, hollow sacs on either side of the prosoma, each of these opening to the outside via one orifice (ozopore) immediately above coxa I. In contrast to the situation known from laniatorean, cyphophthalmid and some eupnoid Opiliones, ozopores are not exposed but hidden in a depression (atrium), formed by a dorsal integumental fold of the carapace and the dorsal parts of coxae I. Glandular sacs are connected to ozopores via a short duct which is equipped with a specific closing mechanism in its distal part: A layer of modified epidermal cells forms a kind of pad-like tissue, surrounding the duct like a valve. Several muscles attached to the anterior parts of the glandular reservoir and to the epithelial pad may be associated with ozopore-opening. The actual mechanism of secretion discharge seems to be highly unusual and may be hypothesized on the basis of corroborating data from behavioral observations, scent gland anatomy and secretion chemistry as follows: Enteric fluid is considered to be directed towards the ozopores via cuticular grooves in the surface of the coxapophyses of legs I. Then, the fluid is sucked into the anterior part of the scent gland reservoirs by the action of dorsal dilator muscles that widen the reservoir and produce a short-term negative pressure. After dilution/solution of the naphthoquinone-rich scent gland contents, a secretion-loaded fluid is thought to be discharged with the help of transversal compressor muscles. This is the first detailed study on the functional anatomy of scent glands and the mechanisms of secretion discharge in the Dyspnoi. PMID- 21618270 TI - Synthesis, cytotoxicity testing, and structure-activity relationships of novel 6 chloro-7-(4-phenylimino-4H-3,1-benzoxazin-2-yl)-3-(substituted)-1,4,2 benzodithiazine 1,1-dioxides. AB - A new series of 16 6-chloro-1,1-dioxo-7-{4-[(4-R(1)-phenyl)imino]-4H-3,1 benzoxazin-2-yl}-3-(substituted amino)-1,4,2-benzodithiazines 7-22 was prepared in order to evaluate the cytotoxic activity against six human cancer cell lines. The structures of the new compounds were confirmed by IR, (1)H-, and (13)C-NMR, elemental analysis and in the cases of 11 and 31 by X-ray crystal structure analysis. This analysis showed that contrary to our earlier report the structures contain a benzoxazine ring instead of the proposed quinazolinone ring. The bioassay indicated that the benzodithiazine derivatives 7-22 possess cancer cell growth-inhibitory properties. Some compounds showed a high level of selectivity for certain cell lines. The most active compounds 11, 12, 16, 19, 21, and 22 exhibited potency higher or comparable to cisplatin. The compounds were particularly effective in LCLC-103H and MCF-7 cell lines with IC(50) values of 0.49-1.60 uM. Quantitative structure activity relationships (QSAR) revealed that a chloro substituent R(1) in the phenyl ring as well as the shape of the substituted amino group at R(2) (e.g., unsaturation is beneficial) are important for potency. PMID- 21618271 TI - Chemistry of phosphorus ylides. Part 33. Synthesis and antitumor activities of some new chromenone derivatives. AB - The reaction of nucleophilic phosphacumulene and phosphallene ylides with different chromenone derivatives was investigated. Heterocycles and carbocycles of various ring sizes and heteroatom patterns such as pyrano-, oxaphosphino-, cyclopenta-, phosphoranylidene cyclobutane-, and phospholo-chromenones were obtained. The antitumor (breast and liver) properties of some new compounds were performed in vitro. Some of these compounds were more potent than the comparative standard. PMID- 21618272 TI - Synthesis and in-vitro antimicrobial activity of secondary and tertiary amines containing 2-chloro-6-methylquinoline moiety. AB - A number of secondary and tertiary amines bearing 2-chloro-6-methylquinoline were synthesized by nucleophilic substitution reaction of 3-(chloromethyl)-2-chloro-6 methylquinoline with substituted aromatic primary and secondary amines in presence of catalytic amount of triethylamine (TEA) and K(2)CO(3). All the compounds were characterized by combined use of IR, (1)H-NMR, (13)C-NMR, mass spectral data, and microanalyses. The newly synthesized quinolinyl amines were screened in vitro for their antifungal activity against Aspergillus niger MTCC 281, Aspergillus flavus MTCC 277, Monascus purpureus MTCC 369, Penicillium citrinum NCIM 768 and for antibacterial activity strains viz. Escherichia coli NCTC 10418, Staphylococcus aureus NCTC 65710, and Pseudomonas aeruginosa NCTC 10662 by agar diffusion technique. Results of the preliminary screening revealed that some of the compounds mainly those with electron withdrawing groups in the phenyl ring showed promising antifungal activity. PMID- 21618273 TI - Quality of life after treatment with midurethral sling and concomitant prolapse repair in patients with mixed versus stress urinary incontinence. AB - AIMS: To compare quality of life in patients with mixed urinary incontinence (MUI) to stress urinary incontinence (SUI) after treatment with a retropubic midurethral sling and concomitant prolapse repair. METHODS: Sixty-three patients met inclusion criteria. Patients completed the short form of the urogenital distress inventory (UDI-6) questionnaire pre- and postoperatively. Concomitant procedures included anterior/posterior colporrhaphy, paravaginal repair, LeForte, vaginal hysterectomy, and sacrospinous ligament fixation. RESULTS: Preoperatively, those with MUI had significantly higher UDI-6 scores (59.8 +/- 16.6 vs. 42.3 +/- 14.5, respectively, P < 0.0001). Postoperatively, there was no difference in UDI-6 scores (P = 0.87). Subjective cure rate for SUI was the same in patients with MUI versus SUI (73%). Patients with MUI demonstrated a significantly greater improvement in UDI-6 scores as compared to patients with SUI. CONCLUSIONS: Patients with MUI compared with SUI demonstrated a greater improvement in quality of life after midurethral sling placement and concomitant pelvic organ prolapse repair as assessed by a disease-specific quality of life questionnaire (UDI-6). PMID- 21618274 TI - Stranger at the gate: the effect of the plaintiff's use of an interpreter on juror decision-making. AB - There exists a substantial literature examining the effect on juror decision making of extraneous demographic characteristics of plaintiffs and defendants. In most of these studies, members of groups that are perceived as being minorities or as belonging to one of a variety of outgroups (lower socioeconomic status, immigrants) are treated more harshly by jurors, or are perceived as being less deserving or credible. In this study, the authors examine treatment by jurors of a relatively less well investigated outgroup: that of the non-English speaker. An experiment was conducted in which actual jurors in a large urban county were randomly assigned to view a videotape of a civil case. Three versions of the videotapes were identical except that, on one, the plaintiff required an interpreter to communicate and it is approximately three minutes longer than the other two. On the other two versions, the plaintiff spoke English, but differed in ethnicity (Hispanic or Anglo). The findings showed that the non-English speaking plaintiff did not fare worse than the English speakers, and, in fact, was awarded higher mean damages than either of the English speakers. PMID- 21618275 TI - Mechanisms of collagen fibril alignment in tendon injury: from tendon regeneration to artificial tendon. AB - The process by which collagen fibrils are aligned following tendon injury remains unknown. Therefore, we analyzed the process of tendon regeneration by transmission electron microscopy, using a film model method. In mice, the Achilles tendon of medial head was transected. On day 3, after only the proximal end of the transected tendon was placed on film and kept in vivo, a translucent substance containing granules, called tendon gel, was secreted. On day 5, the granules assembled in a loose (L) layer, and coalesced tightly in a dense (D) layer, forming an L-D-L layered pattern. On day 10, granules showed high electron density in H layers, which developed into D-H-D layers on day 13. The distal end was placed on film to face the proximal end. On day 10, the tendon gel showed a D H-D layer pattern. On day 11, mechanical stress from muscular constriction changed the tendon gel to aligned collagen fibrils (6 +/- 2 nm in diameter). Thereafter, the diameter of the fibrils increased. Tendon gel harvested on day 5 or day 10 was pulled manually or by hanging weights (about 0.6 MPa). Aligned collagen fibrils (32 +/- 7 nm in diameter) were created by traction using tendon gel harvested on day 10. PMID- 21618276 TI - PDGF in bone formation and regeneration: new insights into a novel mechanism involving MSCs. AB - With the identification of mesenchymal stem cells (MSCs) as pericytes, the details of bone formation, regeneration, and repair take on new meaning. Growth factors and other signaling molecules together with MSCs play important roles in these bone fabrication processes. However, the interaction of these cellular healing components is not completely understood. The formation of new vasculature is critical to regeneration and repair as both the driver and orientor of new bone formation. In this context, MSCs are proposed to be largely derived from pericytes associated with the vasculature. A comprehensive perspective is presented in which signaling molecules such as PDGF take on new significance in the vasculature-pericyte-MSC-osteoblast dynamics. Current data suggest that PDGF could function as a central connector between the cellular components and contributors of the osteoblast differentiation program. The inference is that PDGF could function at sites of injury to mobilize the pericytes from their abluminal location, stimulate mitotic expansion of these cells and help organize them. In this way, PDGF both contributes to the osteogenic lineage and helps to stabilize newly forming vessels that act to drive the multistep, multicomponent cascade of new bone formation. This thesis explains how PDGF functions as a powerful therapeutic agent for bone formation and repair. PMID- 21618277 TI - Reconstruction of composite tibial defect with free flaps and ipsilateral vascularized fibular transposition. AB - Composite defects of the tibia following open fractures are among the most challenging of clinical problems. The aim of this study is to report the results of treatment using a free flap procedure followed by ipsilateral vascularized fibular transposition (IVFT) for reconstruction of composite tibial defects. Ten patients underwent a free flap procedure followed by IVFT and plating. The mean size of the flaps was 12.1 * 6 cm(2). The mean length of bone defect was 5.35 cm. IVFT were performed 4.3 months following the free flap. Patients were followed for an average of 3.4 years. All flaps survived. The average time to union of the proximal and distal ends was 5.2 and 6.7 months, respectively. There were neither stress fractures of the transferred fibula nor recurrent infections. One patient demonstrated a medial angulation of 8 degrees in the reconstructed tibia but experienced no difficulties in activities of daily living. At the last follow-up time point, all patients were able to walk without an assist device and were satisfied with the preservation of the injured lower extremity. Free flap procedures followed by IVFT for the treatment of composite tibial defects may reduce complications at the recipient site and infections, such as osteomyelitis. The plating technique combined with IVFT allowed bone union without additional operations or stress fractures in our series. We suggest that staged free flap and IVFT is useful for the treatment of composite segmental tibial defects. PMID- 21618278 TI - A unique deep inferior epigastric artery perforator and implications for a muscle and fascia sparing vertical rectus abdominis myocutaneous flap: a case report. AB - Despite the sacrifice of rectus abdominis muscle, the vertical rectus abdominis musculocutaneous (VRAM) flap is still a preferred option for perineal reconstruction. This journal has previously reported on the utility of preoperative computed tomographic angiography (CTA) in this setting to identify cases that are both suitable and unsuitable for rectus abdominis flaps after previous surgery. We report a case which highlights a unique example of the benefits of such imaging, with the largest deep inferior epigastric artery (DIEA) perforator described to date identified on imaging, and used to potentiate a donor-site sparing procedure. The use of this dominant perforator was able to limit donor site harvest to only a small cuff of anterior rectus sheath and a small segment of rectus abdominis, potentiating a muscle-sparing and fascia sparing VRAM flap for perineal reconstruction. As such, preoperative CTA was found to be a useful tool in identifying a unique anatomical variant in the largest DIEA perforator described to date, and was used to potentiate a muscle sparing and fascia-sparing VRAM flap for perineal reconstruction. PMID- 21618279 TI - Make no bones about it (regulation and control): NGF versus TGF-beta and BMP. PMID- 21618280 TI - Accumlation of lipoprostaglandin E1 at the anastomotic region after systemic application. AB - Lipoprostaglandin E1 (lipo-PGE1) has been found accumulating in injured vascular regions. This study examined the localization of lipo-PGE1 in the anastomotic region. The study was divided into three parts. First, we performed anastomosis of the rat femoral artery and vein (n = 17). Lipo-PGE1 labeled with 1,1' dioctadecyl-1,3,3',3'-tetramethyl-indocarbocyanine was infused intravenously. Hematoxylin-Eosin staining and fluorescence microscopic findings showed that lipo PGE1 markedly accumulated at the anastomotic site when compared to the contralateral non anastomotic region. Then, we measured laser Doppler flow (LDF) of a lower leg before and after infusion of lipo-PGE1 (n = 7) and saline (n = 7). Increase of blood flow was maintained 1 hour after the infusion of lipo-PGE1 (144% +/- 25.0%) when compared to saline infusion. Finally, we performed immunohistochemical and electron microscopic examinations and found that Lipo PGE1 was incorporated in vascular smooth muscle cells of the anastomotic region. These findings suggest selective accumulation of the lipo-PGE1 in the vascular anastomosis site and affect on the blood flow of repaired vessels. PMID- 21618281 TI - Paper-traction system to assist and facilitate microvascular anastomosis. PMID- 21618282 TI - Why we choose free vascularized fibular grafting for osteonecrosis of the femoral head? PMID- 21618283 TI - Frontotemporal dementia with parkinsonism presenting as posterior cortical atrophy. PMID- 21618284 TI - Association between the prevalence of depression and age in a large representative German sample of people aged 53 to 80 years. AB - OBJECTIVE: The aim of the study was to determine the association between the prevalence of clinically significant depression and age in a large representative sample of elderly German people. METHODS: In the second follow-up (2005-2007) of the ESTHER cohort study, the 15-item geriatric depression scale (GDS-15) as well as a sociodemographic and clinical questionnaire were administered to a representative sample of 8270 people of ages 53 to 80 years. The prevalence of clinically significant depression was estimated using a GDS cut-off score of 5/6. Prevalence rates were estimated for the different age categories. Association between depression and age was analyzed using logistic regression, adjusted for gender, co-morbid medical disorders, education, marital status, physical activity, smoking, self-perceived cognitive impairment, and anti-depressive medication. RESULTS: Of the participants, 7878 (95.3%) completed more than twelve GDS items and were included in the study. The prevalence of clinically significant depression was 16.0% (95%CI = [15.2; 16.6]). The function of depression prevalence dependent on age group showed a U-shaped pattern (53-59: 21.0%, CI = [18.9; 23.3]; 60-64: 17.7%, CI = [15.7; 19.7]; 65-69: 12.6%, CI = [11.2; 14.0]; 70-74: 14.4%, CI = [12.6; 16.0]; 75-80: 17.1%, CI = [14.9; 19.4]). Adjusted odds ratios showed that the chances of being depressive decrease with the age category but remain relatively stable for people aged 65 and over. CONCLUSIONS: The prevalence of depression in the elderly seems to be associated with the age category. Adjusted odds ratios showed that people aged 60 and older had lower chances of being depressive than people aged 53 to 59 years. PMID- 21618285 TI - Staging of the cognitive decline in Alzheimer's disease: insights from a detailed neuropsychological investigation of mild cognitive impairment and mild Alzheimer's disease. AB - OBJECTIVE: The decline of episodic memory in Alzheimer's disease (AD) is well established, but the exact appearance and staging of deficits in other cognitive domains is sometimes contentious. The current investigation attempted to elucidate the appearance of additional cognitive deficits in the non-episodic domains and to understand these deficits with respect to the known pathological staging of AD. METHODS: A cross-sectional investigation compared cognitively normal age-matched controls with patients with mild AD and mild cognitive impairment (MCI) using a detailed neuropsychological assessment. RESULTS: The systematic investigation of cognitive performance across the major cognitive domains demonstrates that the appearance of additional cognitive deficits in MCI and AD can be predicted, with impaired semantic cognition performance pre-empting the appearance of attention/executive dysfunction and visuospatial deficits in the majority of patients with MCI. CONCLUSIONS: This progressive pattern of cognitive deficits fits with the known pathological staging of AD, and the data further highlight the relative rarity of pure amnestic MCI. These results indicate that any neuropsychological test battery used to assess patients with MCI should include language and semantic memory tests in addition to typical episodic memory tests, as changes within this domain might be a sensitive indication of incipient AD. PMID- 21618286 TI - Alcohol diagnoses among older Tennessee Medicare beneficiaries: race and gender differences. AB - BACKGROUND: These analyses bolster a sparse body of research focusing on the rate of alcohol disorders among older adults, particularly race and gender subgroups. METHODS: We based the study on cross-sectional data from all Medicare billed physician/patient encounters. Analyses of these data included cross-tabulations, difference of means tests, and difference of proportions tests, logistic regression and multinomial logistic regression. These analyses were based Medicare billing records from physician/patient encounters in Tennessee. Data included Tennessee Medicare billings beneficiaries enrolled in Medicare Part B, who saw a physician at least once in 2000. Patients with billings containing ICD 9 codes: 303 (alcohol abuse), 305 (alcohol dependence), 291 (alcohol psychosis), or 571.1-571.3 (alcohol-related liver disease including cirrhosis of the liver) as to primary diagnosis were considered alcohol-disordered. RESULTS: Analyses reveal the overall rate of alcohol disorders, subgroup variation in rates and differences in pattern of specific disorders. Merely 0.04% of Tennessee Medicare beneficiaries were diagnosed with any type of alcoholism, a rate much lower than those reported in previous studies. Rates of alcohol disorders varied across groups, with significantly higher rates for Black men. The type alcohol disorder also varied across groups. CONCLUSIONS: Many encounters with the medical system result in missed opportunities to identify and treat alcohol disorders, a significant risk factor among older adults. Alcoholism both triggers and exacerbates many chronic conditions among older adults. The earlier in the disease trajectory the more of these conditions could be prevented or more efficiently managed, resulting in substantial savings in health care costs. PMID- 21618287 TI - Semantic organizational strategy predicts verbal memory and remission rate of geriatric depression. AB - OBJECTIVE: This study tests the hypothesis that the use of semantic organizational strategy during the free-recall phase of a verbal memory task predicts remission of geriatric depression. METHODS: Sixty-five older patients with major depression participated in a 12-week escitalopram treatment trial. Neuropsychological performance was assessed at baseline after a 2-week drug washout period. The Hopkins Verbal Learning Test-Revised was used to assess verbal learning and memory. Remission was defined as a Hamilton Depression Rating Scale score of <= 7 for 2 consecutive weeks and no longer meeting the DSM-IV-TR criteria for major depression. The association between the number of clusters used at the final learning trial (trial 3) and remission was examined using Cox's proportional hazards survival analysis. The relationship between the number of clusters utilized in the final learning trial and the number of words recalled after a 25-min delay was examined in a regression with age and education as covariates. RESULTS: Higher number of clusters utilized predicted remission rates (hazard ratio, 1.26 (95% confidence interval, 1.04-1.54); chi(2) = 4.23, df = 3, p = 0.04). There was a positive relationship between the total number of clusters used by the end of the third learning trial and the total number of words recalled at the delayed recall trial (F(3,58) = 7.93; p < 0.001). CONCLUSIONS: Effective semantic strategy use at baseline on a verbal list learning task by older depressed patients was associated with higher rates of remission with antidepressant treatment. This result provides support for previous findings indicating that measures of executive functioning at baseline are useful in predicting antidepressant response. PMID- 21618288 TI - Teacher-delivered resilience-focused intervention in schools with traumatized children following the second Lebanon War. AB - The 2006 Lebanon War exposed children in the north of Israel to daily rocket attacks. To cope with the massive psychological needs, a teacher-delivered protocol focusing on enhancing personal resilience was implemented. Children were assessed for risk factors, symptoms, and adaptation before the 16-week program (Time 1; n = 983) and after its completion (Time 2; n = 563). At a 3-month follow up (Time 3; n = 754) children were assessed together with a waiting-list comparison group (n = 1,152). Participating children showed a significant symptom decrease at Time 2 and significantly fewer symptoms than the control group at Time 3. Six or more risk factors were associated with greater symptoms and parental concern about the child's adaptive functioning. Teachers are valuable cost-effective providers for clinically informed interventions after mass trauma and disaster. PMID- 21618289 TI - Avoidant coping as a mediator between peritraumatic dissociation and posttraumatic stress disorder symptoms. AB - Peritraumatic dissociation consistently predicts posttraumatic stress disorder (PTSD). Avoidant coping may serve as a mechanism through which peritraumatic dissociation contributes to PTSD symptoms. Path analysis was used to examine whether avoidant coping assessed 6 weeks following a motor vehicle accident mediated the relationship between in-hospital peritraumatic dissociation and 6 month (n = 193) and 12-month (n = 167) chronic PTSD symptoms. Results revealed that, after controlling for age, gender, depression, and 6-week PTSD symptoms, avoidant coping remained a partial mediator between peritraumatic dissociation and chronic PTSD symptoms 6- and 12-months postaccident. Post-hoc multigroup analyses suggested that at 6-months posttrauma, the mediation was significant in women, but not in men. Gender-specific results were not significant at 12-months posttrauma. Interventions targeted at reducing avoidant coping in high dissociators may aid in reducing PTSD symptoms. PMID- 21618290 TI - Relationships between soldiers' PTSD symptoms and spousal communication during deployment. AB - Social support, including support from spouses, may buffer against posttraumatic stress disorder (PTSD) symptoms. The current study assessed whether the frequency of spousal communication during a recent deployment, a potentially important source of support for soldiers, was related to postdeployment PTSD symptoms. Data came from 193 married male Army soldiers who returned from military deployment within the past year. For communication modalities conceptualized as delayed (i.e., letters, care packages, and e-mails), greater spousal communication frequency during deployment was associated with lower postdeployment PTSD symptom scores, but only at higher levels of marital satisfaction (p = .009). At lower marital satisfaction, more delayed spousal communication during deployment was associated with more PTSD symptoms (p = .042). For communication modalities conceptualized as interactive (i.e., phone calls, instant messaging, instant messaging with video), the same general direction of effects was seen, but the interaction between communication frequency and marital satisfaction predicting PTSD symptoms did not reach significance. PMID- 21618291 TI - Pattern transfer and electric-field-induced magnetic domain formation in multiferroic heterostructures. PMID- 21618292 TI - Nanoscale electronics: digital fabrication by direct femtosecond laser processing of metal nanoparticles. PMID- 21618293 TI - Triggered cell release from materials using bioadhesive photocleavable linkers. PMID- 21618294 TI - Depleted bulk heterojunction colloidal quantum dot photovoltaics. PMID- 21618295 TI - Multi-path magnetic relaxation of mono-dysprosium(III) single-molecule magnet with extremely high barrier. PMID- 21618296 TI - Spontaneous transmission of chirality through multiple length scales. AB - The hierarchical transfer of chirality in nature, from the nano-, to meso-, to macroscopic length scales, is very complex, and as of yet, not well understood. The advent of scanning probes has allowed chirality to be monitored at the single molecule or monolayer level and has opened up the possibility to track enantiospecific interactions and chiral self-assembly with molecular-scale detail. This paper describes the self-assembly of a simple, model molecule (naphtho[2,3-a]pyrene) that is achiral in the gas phase, but becomes chiral when adsorbed on a surface. This polyaromatic hydrocarbon forms a stable and reversibly ordered system on Cu(111) in which the transmission of chirality from single surface-bound molecules to complex 2D chiral architectures can be monitored as a function of molecular packing density and surface temperature. In addition to the point chirality of the surface-bound molecule, the unit cell of the molecular domains was also found to be chiral due to the incommensurate alignment of the molecular rows with respect to the underlying metal lattice. These molecular domains always aggregated in groups of three, all of the same chirality, but with different rotational orientations, forming homochiral "tri lobe" ensembles. At a larger length scale, these tri-lobe ensembles associated with nearest-neighbor tri-lobe units of opposite chirality at lower packing densities before forming an extended array of homochiral tri-lobe ensembles at higher converges. This system displayed chirality at a variety of size scales from the molecular (~1 nm) and domain (~5 nm) to the tri-lobe ensemble (~10 nm) and extended array (>25 nm) levels. The chirality of the tri-lobe ensembles dictated how the overall surface packing occurred and both homo- and heterochiral arrays could be reproducibly and reversibly formed and interchanged as a function of surface coverage. Finally, these chirally templated surfaces displayed remarkable enantiospecificity for naphtho[2,3-a]pyrene molecules adsorbed in the second layer. Given its simplicity, reversibility, and rich degree of order, this system represents an ideal test bed for the investigation of symmetry breaking and the hierarchical transmission of chirality through multiple length scales. PMID- 21618297 TI - Solvent and TMEDA effects on the configurational stability of chiral lithiated aryloxiranes. AB - The employment of hexane/N,N,N',N'-tetramethylethylenediamine (TMEDA) dramatically hinders the racemization of those lithiated styrene oxides (trifluoromethyl-, chloro-, and phenylthio-substituted) that have been proven to be configurationally unstable in THF on the timescale of their reactions. The barriers to inversion and the activation parameters, calculated (Eyring equation) for reactions performed in THF, THF/TMEDA, and hexane/TMEDA, suggest the intervention of particular enantiomerization mechanisms for each case. The role of TMEDA in both coordinating and noncoordinating solvents has also been questioned and discussed in light of the kinetic data gathered and a model for deprotonation in hexane/TMEDA has also been proposed. The synthetic benefits of our results became apparent on establishing an asymmetric synthesis of an industrially important antifungal agent. PMID- 21618298 TI - Iterative dianion relay along the ring: formation of gem-bis(trimethylsilyl) cyclopentenones from 2,5-bis(trimethylsilyl) oxy-cyclopentadienyl dianions and acid chlorides. PMID- 21618299 TI - Expeditious assembly of a 2-amino-4H-chromene skeleton by using an enantioselective Mannich intramolecular ring cyclization-tautomerization cascade sequence. PMID- 21618300 TI - A [2]catenane displaying pirouetting motion triggered by debenzylation and locked by chloride anion recognition. PMID- 21618301 TI - Rh(III)-catalyzed oxidative olefination of vinylic C-H bonds: efficient and selective access to di-unsaturated alpha-amino acid derivatives and other linear 1,3-butadienes. PMID- 21618302 TI - Potentiation of antimicrobial activity of ciprofloxacin by Pelargonium graveolens essential oil against selected uropathogens. AB - The recent approach of using herbs and antibiotics in combination constitutes a strategy to overcome the problems of resistance and side effects associated with conventional antibiotics. In the present study, the antimicrobial effect of Pelargonium graveolens L' Her essential oil in combination with ciprofloxacin was evaluated on uropathogens, namely, Klebsiella pneumoniae KT2, Proteus mirabilis PRT3 and Staphylococcus aureus ST2. Minimum inhibitory concentrations of P. graveolens essential oil and ciprofloxacin were determined by the microbroth dilution method and further, the interaction between these two agents was studied by a checkerboard method. The fractional inhibitory concentration index (FICI) was calculated to be 0.375 for both K. pneumoniae KT2 and P. mirabilis PRT3, while for S. aureus ST2 it was found to be 0.5. The values of FICI for the tested microorganisms were found to be <=0.5, which indicates synergism between P. graveolens essential oil and ciprofloxacin. The concave shaped curve in the isobolograms also depicted a synergistic effect of P. graveolens essential oil and ciprofloxacin against the tested microorganisms. Hence, the synergistic action of P. graveolens essential oil and ciprofloxacin may be applied for the treatment of UTIs, which have hitherto been treated by using only synthetic drugs. PMID- 21618303 TI - Diallyl disulfide induces MUC5B expression via ERK2 in human airway epithelial cells. AB - Garlic has been shown to have antimicrobial, hypolipidemic, antithrombotic, antitumor and immunostimulatory properties. The medicinal effects of garlic are derived from the flavonoid and organosulfur components. Diallyl disulfide (DADS), an organosulfur, is the main component responsible for the diverse biological effects of garlic. However, the effects of DADS on mucin gene expression in airway epithelial cells have not been reported to date. Therefore, this study was performed to investigate the effects and brief signaling pathway of DADS associated with MUC5B expression in NCI-H292 epithelial cells using RT-PCR, ELISA, western blot, immunocytochemistry and cell transfection with siRNA. DADS induced MUC5B expression and activated the phosphorylation of ERK1/2 MAPK. In addition, U0126 inhibited DADS-induced MUC5B expression and DADS-activated phosphorylation of ERK1/2 MAPK. Moreover, the immunopositive cells for MUC5B protein did not appear after treatment of DADS with U0126, and the knockdown of ERK2 MAPK by ERK2 MAPK siRNA significantly blocked DADS-induced MUC5B mRNA expression. However, DADS did not activate the phosphorylation of p38 MAPK, and SB203580 did not inhibit DADS-induced MUC5B expression. This is the first study to show that DADS-induced MUC5B expression appears to be regulated by activation of the ERK2 MAPK signaling pathway in human NCI-H292 airway epithelial cells. PMID- 21618304 TI - Diffusion tensor imaging of diffuse axonal injury in a rat brain trauma model. AB - Diffusion tensor imaging (DTI) was used to study traumatic brain injury. The impact-acceleration trauma model was used in rats. Here, in addition to diffusivities (mean, axial and radial), fractional anisotropy (FA) was used, in particular, as a parameter to characterize the cerebral tissue early after trauma. DTI was implemented at 7 T using fast spiral k-space sampling and the twice-refocused spin echo radiofrequency sequence for eddy current minimization. The method was carefully validated on different phantom measurements. DTI of a trauma group (n = 5), as well as a sham group (n = 5), was performed at different time points during 6 h following traumatic brain injury. Two cerebral regions, the cortex and corpus callosum, were analyzed carefully. A significant decrease in diffusivity in the trauma group versus the sham group was observed, suggesting the predominance of cellular edema in both cerebral regions. No significant FA change was detected in the cortex. In the corpus callosum of the trauma group, the FA indices were significantly lower. A net discontinuity in fiber reconstructions in the corpus callosum was observed by fiber tracking using DTI. Histological analysis using Hoechst, myelin basic protein and Bielschowsky staining showed fiber disorganization in the corpus callosum in the brains of the trauma group. On the basis of our histology results and the characteristics of the impact-acceleration model responsible for the presence of diffuse axonal injury, the detection of low FA caused by a drastic reduction in axial diffusivity and the presence of fiber disconnections of the DTI track in the corpus callosum were considered to be related to the presence of diffuse axonal injury. PMID- 21618305 TI - Association between sociability and diffusion tensor imaging in BALB/cJ mice. AB - The purpose of this study was to use high-resolution diffusion tensor imaging (DTI) to investigate the association between DTI metrics and sociability in BALB/c inbred mice. The sociability of prepubescent (30-day-old) BALB/cJ mice was operationally defined as the time that the mice spent sniffing a stimulus mouse in a social choice test. High-resolution ex vivo DTI data on 12 BALB/cJ mouse brains were acquired using a 9.4-T vertical-bore magnet. Regression analysis was conducted to investigate the association between DTI metrics and sociability. Significant positive regression (p < 0.001) between social sniffing time and fractional anisotropy was found in 10 regions located in the thalamic nuclei, zona incerta/substantia nigra, visual/orbital/somatosensory cortices and entorhinal cortex. In addition, significant negative regression (p < 0.001) between social sniffing time and mean diffusivity was found in five areas located in the sensory cortex, motor cortex, external capsule and amygdaloid region. In all regions showing significant regression with either the mean diffusivity or fractional anisotropy, the tertiary eigenvalue correlated negatively with the social sniffing time. This study demonstrates the feasibility of using DTI to detect brain regions associated with sociability in a mouse model system. PMID- 21618306 TI - Lactate MRSI and DCE MRI as surrogate markers of prostate tumor aggressiveness. AB - Longitudinal studies of lactate MRSI and dynamic contrast-enhanced MRI were performed at 4.7 T in two prostate tumor models grown in rats, Dunning R3327-AT (AT) and Dunning R3327-H (H), to determine the potential of lactate and the perfusion/permeability parameter Ak(ep) as markers of tumor aggressiveness. Subcutaneous AT (n = 12) and H (n = 6) tumors were studied at different volumes between 100 and 2900 mm(3) (Groups 1-5). Lactate concentration was determined using selective multiple quantum coherence MRSI with the phantom substitution method. Tumor enhancement after the administration of gadolinium diethylenetriaminepenta-acetic acid was analyzed using the Brix-Hoffmann model and the Ak(ep) parameter was used as a measure of tumor perfusion/permeability. Lactate was not detected in the smallest AT tumors (Group 1; 100-270 mm(3) ). In larger AT tumors, the lactate concentration increased from 2.8 +/- 1.0 mm (Group 2; 290-700 mm(3)) to 8.4 +/- 2.9 mm (Group 3; 1000-1340 mm(3)) and 8.2 +/- 2.2 mm (Group 4; 1380-1750 mm(3) ), and then decreased to 5.0 +/- 1.7 mm (Group 5; 1900 2500 mm(3)), and was consistently higher in the tumor core than in the rim. Lactate was not detected in any of the H tumors. The mean tumor Ak(ep) values decreased with increasing volume in both tumor types, but were significantly higher in H tumors. In AT tumors, the Ak(ep) values were significantly higher in the rim than in the core. Histological hypoxic and necrotic fractions in AT tumors increased with volume from 0% in Group 1 to about 20% and 30%, respectively, in Group 5. Minimal amounts of hypoxia and necrosis were found in H tumors of all sizes. Thus, the presence of lactate and heterogeneous perfusion/permeability are signatures of aggressive, metabolically deprived tumors. PMID- 21618307 TI - Classification of Greek Mentha pulegium L. (Pennyroyal) samples, according to geographical location by Fourier transform infrared spectroscopy. AB - INTRODUCTION: Mentha pulegium L. (pennyroyal) is one of the four most commercially important Mentha species, even it is not a cultivated plant. It can be abundantly located in the Iberian Peninsula and North African countries. In Greece it grows in the wild and it is scattered all over the country. Pennyroyal is best known for its essential oil, with Spain and Morocco being the largest producers in the world. Mid-infrared spectroscopy has been applied to determine the origin of various samples. OBJECTIVES: In this work Fourier transform infrared spectroscopy (FT-IR) combined with canonical discriminant analysis has been applied to distinguish 70 Greek pennyroyal samples according to their collection areas. MATERIAL AND METHODS: Pennyroyal nonpolar organic extracts were prepared using ultrasound-assisted solvent extraction. The spectra of the extracts were recorded in the range of 4000-400 cm(-1) and the best discrimination was achieved in the spectral region 1720-1650 cm(-1) . RESULTS: Spectral features for the discrimination of pennyroyal samples among the different collection areas occur primarily in the carbonyl region and are correlated with the main volatile constituents of the extracts (menthone, isomenthone, pulegone, piperitone). All areas were easily differentiated by canonical discriminant analysis. The percentages of correct classification and validation were 94.3 and 90.0%, respectively. CONCLUSION: The combination of FT IR spectroscopy and multivariate analysis provides a rapid and ambient method to discriminate pennyroyal samples in terms of geographical origin. PMID- 21618309 TI - A new method for the simultaneous analysis of strychnine and brucine in Strychnos nux-vomica unprocessed and processed seeds using a carbon-paste electrode modified with multi-walled carbon nanotubes. AB - INTRODUCTION: Strychnos nux-vomica L. (Loganiaceae), widely used in folk medicine, is grown extensively in southern Asian countries. Its major bioactive constituents are strychnine and brucine, which are frequently used in traditional herbal medicines for treatment of nervous diseases, vomiting and traumatic pain. OBJECTIVE: A new method using a carbon-paste electrode modified with multi-walled carbon nanotubes (CNT/CPE) was developed and validated for single or simultaneous determination of strychnine and brucine in Strychnos nux-vomica seeds. Additionally, an environmentally friendly method was successfully applied to reduce the levels of strychnine and brucine in seeds. MATERIALS AND METHODS: Cyclic voltammetry, chronocoulometry and differential pulse voltammetry were used with multi-walled carbon nanotube modified carbon-paste electrodes. RESULTS: The peak currents increase linearly with the strychnine and brucine concentrations in the ranges of 50-1000 and 5-355 u m, and the detection limits for strychnine and brucine were 0.43 and 0.28 u m, respectively. Of the processing methods used, the greatest reduction in the strychnine and brucine levels was observed in samples processed using milk and saltwater. CONCLUSION: A new, sensitive and selective method was developed for the measurement of strychnine and brucine. This method was successfully applied to the determination of strychnine and brucine in unprocessed and processed Strychnos nux-vomica seed. PMID- 21618308 TI - A comparison of headspace solid-phase microextraction and classic hydrodistillation for the identification of volatile constituents from Thapsia spp. provides insights into guaianolide biosynthesis in Apiaceae. AB - INTRODUCTION: Thapsia spp. (Apiaceae) are the major natural source of polyoxygenated guaianolide sesquiterpene lactones known as thapsigargins, which induce apoptosis in mammalian cells via a high affinity inhibition of the sarco/endoplasmic reticulum Ca(2+) ATPase. The mechanism of biosynthesis of thapsigargins has not been elucidated, and probable biochemical precursors such as hydrocarbon or oxygenated sesquiterpenes have not been identified in previous phytochemical analyses of essential oils from this genus. OBJECTIVE: To investigate the utility of solid phase micro-extraction (SPME), when compared with classical essential oil distillates, for identifying potential precursors of guaianolide sesquiterpene lactones from Thapsia garganica L. and Thapsia villosa L. type II. METHODOLOGY: A systematic description of the volatile components of roots, flowers, stems and fruits of T. villosa and of root, flower and fruits of T. garganica was constructed via GC-MS analyses of SPME-adsorbed compounds and of essential oils obtained through hydrodistillation of the same tissues. RESULTS: The sesquiterpenoids delta-cadinene, alpha- and delta-guaiene, elemol and guaiols were found to be major volatile constituents of the roots of T. garganica and T. villosa trapped using SPME. In contrast, these sesquiterpenoids were not detected or were at negligible levels in essential oils, where sesquiterpenoids are potentially converted to azulenes during hydrodistillation. CONCLUSION: The new data reported in this study demonstrates that SPME is a valuable tool for the identification of volatile sesquiterpenes when compared with analysis of essential oils, and we postulate that guaiene is the likely precursor of guaianolide sesquiterpenes from Thapsia. PMID- 21618310 TI - Effect of high relative humidity on dried Plantago lanceolata L. leaves during long-term storage: effects on chemical composition, colour and microbiological quality. AB - INTRODUCTION: Modern phytotherapy and quality assurance requires stability data on bioactive metabolites to identify and minimise decomposing factors during processing and storage. A compound's stability in a complex matrix can be different from the stability of the purified compound. OBJECTIVE: To test the stability of iridoids and acteoside and quantify changes in colour and microbiological quality in a common herbal tea, dried P. lanceolata leaves during exposure to high-humidity air. To test the contribution of fungi to metabolite decomposition. METHODOLOGY: Dried P. lanceolata leaves were exposed to atmospheres of different relative humidity (75, 45 and 0%) for 24 weeks. Changes in aucubin and catalpol concentration were determined by CE-MEKC, and those in acteoside on TLC. Colour and chlorophyll-like pigments were measured by different spectrophotometric methods. The number of fungi was monitored; 10 strains were isolated from the plant drug, and their ability to decompose the analytes of interest was tested. RESULTS: During incubation at 75% relative humidity (RH), aucubin, catalpol and acteoside concentrations decreased by 95.7, 97.0 and 70.5%, respectively. Strong shifts were detected in CIELAB parameters a* and b* (browning) as a result of conversion of chlorophyll to pheophytin. Intensive microbial proliferation was also observed. Changes at 45 or 0% RH were typically insignificant. Seven of the 10 isolated fungal strains could decompose both iridoids, and five could decompose acteoside in vitro. CONCLUSION: It was shown that exposure to water results in loss of bioactive molecules of P. lanceolata dried leaves, and that colonising fungi are the key contributors to this loss. PMID- 21618311 TI - Simultaneous determination of eight anthraquinones in Semen Cassiae by HPLC-DAD. AB - INTRODUCTION: Semen Cassiae (SC), a traditional Chinese herbal medicine for the treatment of various diseases, is known to contain active anthraquinone ingredients. However, since the content of some anthraquinones is too low, previous analytical methods only allow the quantitation of a few anthraquinones or a hydrolysis step has to be included in the sample preparation. A rapid and accurate method to examine the content of as many anthraquinones as possible in SC would be desirable. OBJECTIVE: To develop a rapid, sensitive and accurate high performance liquid chromatography-diode array detection (HPLC- DAD) method to simultaneously quantify eight major anthraquinones (obtusifolin-2-glucoside, aurantio-obtusin, aloe- emodin, rhein, obtusifolin, emodin, chrysophanol and physcion) in SC. METHODOLOGY: The separation of anthraquinones was achieved on a C18-column with a gradient elution using acetonitrile and 0.1% phosphoric acid. The detection wavelength was 278 nm and the analysis finished within 25 min. RESULTS: The limits of detection of these compounds ranged from 0.07 to 0.15 ug/mL while the limits of quantitation ranged from 0.24 to 0.51 ug/mL. All calibration curves showed good linearities (r2 > 0.999) within the test ranges. This validated method was successfully used to analyse 22 batches of SC samples collected from various geographical locations. CONCLUSION: The method was validated to be simple, rapid, accurate and reliable to simultaneously determine eight major anthraquinones in SC. Meanwhile, a more specific anthraquinone, obtusifolin, was proposed to serve as a marker for SC, replacing chrysophanol. PMID- 21618312 TI - Safe use of Doppler ultrasound during the 11 to 13 + 6-week scan: is it possible? PMID- 21618313 TI - ISUOG statement on the safe use of Doppler in the 11 to 13 +6-week fetal ultrasound examination. PMID- 21618314 TI - Effect of maternal betamethasone on fetal and uteroplacental blood flow velocity waveforms. AB - OBJECTIVES: To investigate the effects of antenatal betamethasone on fetal and uteroplacental hemodynamics. METHODS: The study comprised 33 women with singleton high-risk pregnancies (23-33 weeks; 27 pregnancies < 30 weeks) not in labor, but at risk for preterm delivery based on fetal or maternal indications. They were treated with two doses of 12 mg betamethasone intramuscularly 24 h apart to enhance fetal lung maturity. Flow velocity waveforms were recorded with Doppler ultrasound from the umbilical artery, the fetal middle cerebral artery, the ductus venosus and both maternal uterine arteries, once before and twice after betamethasone administration. RESULTS: Twenty-one (64%) women delivered within 4 days, nine (27%) women within 5-7 days and three (9%) within 8-15 days after the first dose of betamethasone. Two days after betamethasone, a decrease in pulsatility index was found in the umbilical artery (P = 0.0002) and ductus venosus (P = 0.003). Changes in the umbilical artery waveform from reversed to absent, and from absent to positive diastolic flow, were noted in 12 of 15 cases (P < 0.01). After 4 days, umbilical artery and ductus venosus velocity waveforms in the undelivered fetuses either returned to the type of waveform observed before treatment or showed further deterioration. No significant effects of betamethasone were observed in the fetal middle cerebral artery and uteroplacental circulation. CONCLUSIONS: Maternal antenatal betamethasone resulted in a significant transient change in the velocity waveform and a decrease in the pulsatility index in the umbilical artery and ductus venosus, but did not influence uteroplacental circulation. These findings indicate a direct effect of betamethasone on fetal circulation. PMID- 21618315 TI - Prenatal management of congenital infantile fibrosarcoma: unexpected outcome. AB - We report the case of a large leg tumor in a fetus at 36 weeks of gestation. The tumor's location, ultrasound characteristics and magnetic resonance imaging (MRI) findings enabled a prenatal diagnosis of congenital fibrosarcoma and allowed us to establish the prognosis. After multidisciplinary discussion, it was decided that the pregnancy should continue to 38 weeks, with surgery or neo-adjuvant chemotherapy scheduled after delivery. Unfortunately, the tumor unexpectedly burst in utero and the newborn died of the consequences of hemorrhagic shock, despite rapid amputation. Histological examination confirmed the nature of the tumor. The prognosis of congenital fibrosarcoma is generally good when there are no metastases. MRI is key to establishing a prenatal diagnosis. However, rupture of the tumor can result in fatal hemorrhaging and the pregnancy should be monitored closely before scheduled delivery. PMID- 21618316 TI - Accuracy of sonographic estimation of fetal head circumference. PMID- 21618318 TI - Mothers in recovery: an occupational perspective. AB - The purposes of this study were to discover the stories of mothers regarding their journeys to addiction and through recovery, to explore the impact of addiction on the occupational performance of mothers and to identify the factors perceived by these mothers as important in their treatment. A narrative inquiry with thematic analysis of data was utilized; semi-structured interviews were conducted with 10 women in treatment for alcohol or other drug addiction who were all mothers of minor children. Alterations in occupational identity, occupational performance patterns and performance capacity were revealed. Environmental elements contributing to addiction and those important in recovery were identified and included structure (or the lack of it) and the persons populating the social environment. This qualitative study is not generalizable and should be viewed as relevant to these informants in this context; participants had histories of chronic substance addiction and are not representative of all addictions patients. Findings can be viewed within the context of other research to help enrich the reader's understanding of the complexity of issues. Further research is recommended to test the efficacy of occupational therapy interventions for substance addictions. Possible intervention studies might include those focused on environmental structure, exploration of interests in treatment and co-occupations of mother and child. PMID- 21618319 TI - Working conditions of occupational therapists in Jordan. AB - The purpose of this study was to investigate the working conditions and factors related to job satisfaction among Jordanian occupational therapists. A self administered survey consisting of several questions about the participants' jobs was developed for this study. The inclusion criteria included occupational therapists who worked in the field of occupational therapy (OT) in Jordan and who had at least six months of experience. The survey was distributed to 120 occupational therapists in different OT settings. One hundred and one occupational therapists from different hospitals, centres, schools and universities responded to the survey. Salaries of the participants were low in comparison with the salaries of the rest of healthcare practitioners in Jordan. Chi-square analysis revealed significant relationship between participants' perceived job satisfaction and the administration of the setting, and between the choice of studying OT and the salary that the participant makes. Most occupational therapists in Jordan are young, have modest experience and work in general OT services in government-owned facilities or paediatric centres. A sense of dissatisfaction was obvious among the participants. Further studies are needed to explore reasons of work satisfaction and dissatisfaction among occupational therapists in Jordan. PMID- 21618320 TI - Optically triggered dissociation of kinetically stabilized block copolymer vesicles in aqueous solution. AB - We demonstrate a strategy for using an optical stimulus to trigger the dissociation of block copolymer (BCP) vesicles in aqueous solution. The BCP, comprising hydrophilic poly(ethylene oxide) (PEO) and a block of poly(methacrylic acid) bearing a number of spiropyran methacrylate comonomer units (P(MAA-co SPMA)), was allowed to firstly self-assemble into large vesicles in aqueous solution at pH=3 with protonated carboxylic acid groups, and then become kinetically stable at pH=8 due to the glassy vesicle membrane of P(MAA-co-SPMA). Fast dissociation of the vesicles was achieved through a cascade of events triggered by UV-induced isomerization from neutral spiropyran to charged merocyanine in the membrane. PMID- 21618321 TI - Fabrication of nanofillers into a granular "nanosupport" for Ziegler-Natta catalysts: towards scalable in situ preparation of polyolefin nanocomposites. AB - This communication reports a strategy for scale-up of an in situ polymerization technique for polyolefin-based nanocomposites preparation, taking layered silicate (clay) and multi-walled carbon nanotubes (MWCNTs) as examples of nanofillers. The strategy is realized by transforming the nanofillers into granular "nanosupports" for Ziegler-Natta catalysts. With a catalyst to polymer replication effect on particle morphology, the in situ prepared nanocomposites are of controlled granular particle morphology. With the polymer particle morphology controlled, the in situ polymerization technique becomes suitable for industrial olefin polymerization processes for mass production of polyolefin nanocomposites. PMID- 21618322 TI - Polyglycidol based amphiphilic double-comb copolymers and their self-association in aqueous solution. AB - Herein, we report synthesis and self-association properties of amphiphilic double comb polymers with polyglycidol backbones. First, a bifunctional polyglycidol precursor is synthesized via monomer activated anionic polymerization. Next, two efficient and orthogonal polymer analogous reactions are carried out for grafting hydrophilic oligoethylene glycol side chains and hydrophobic linear aliphatic side-chains. The polymers are analyzed by means of NMR, GPC, and DSC. From the DSC analysis of the bulk samples it is evident that aliphatic side chains segregate from the polar backbone and thus crystallize. Furthermore, in aqueous media the double-comb polymers spontaneously self-assemble to form a multilayer structure. The present results pave a way to tailor and design amphiphilic polymers based on glycidols. Major advantages are spontaneous self-assembly in water and the possibility to form onion polymersomes relevant to encapsulation. PMID- 21618323 TI - pH-disintegrable polyelectrolyte multilayer-coated mesoporous silica nanoparticles exhibiting triggered co-release of cisplatin and model drug molecules. AB - We report on the fabrication of pH-disintegrable polyelectrolyte multilayer coated mesoporous silica nanoparticles (MSN) capable of triggered co-release of cisplatin and model drug molecules. The outer polyelectrolyte multilayer was assembled from permanently cationic polyelectrolyte, poly(allyl amine hydrochloride) (PAH), and negatively charged polyelectrolyte, P(DMA-co-TPAMA), consisting of N,N-dimethylacrylamide (DMA) and 3,4,5,6-tetrahydrophthalic anhydride-functionalized N-(3-aminopropyl)methacrylamide (TPAMA) monomer units, which exhibits pH-induced charge conversion characteristics. Thus, the subtle alteration of solution pH from 7.4 to ~5-6 can lead to the disintegration of outer polyelectrolyte multilayers, accompanied with the co-release of cisplatin and RhB. PMID- 21618324 TI - The effect of hydrogen bonding on intramolecular chain transfer in polymerization of acrylates. AB - Propagation rate coefficients (k(p) ) for 2-hydroxyethyl acrylate (HEA) have been determined by pulsed-laser polymerization (PLP) combined with size-exclusion chromatography (SEC) between 20 and 60 degrees C using pulse repetition rates of 50 and 100 Hz. The success of PLP-SEC under these conditions suggests that HEA is not subjected to the intramolecular chain transfer to polymer (backbiting) reactions dominant for other acrylates; (13) C NMR analysis shows that the quaternary carbon observed in PLP-generated poly(butyl acrylate) (pBA) samples is not observed in pHEA. These results are related to H-bonding in the system, as it is shown that the introduction of H-bonding by addition of n-butanol to BA suppresses backbiting, and the disruption of H-bonding by addition of dimethylformamide to HEA leads to an increased level of backbiting. PMID- 21618326 TI - Diagnosis and management of parotid carcinoma with a special focus on recent advances in molecular biology. AB - Recent progress in diagnosis, treatment, prognosis, and outcome of parotid cancer is reviewed. Modern imaging allows evaluation of the anatomical extent of the cancer and its relationship to the facial nerve, and the World Health Organization (WHO) Histological Classification facilitates accurate, consistent diagnosis. Surgery remains the treatment of choice with preservation of a functioning facial nerve. Resection of the facial nerve should only be undertaken when there is clinical evidence of nerve dysfunction. The NO neck should be treated in advanced-stage and high-grade cancers, but the choice between elective surgery and elective irradiation remains controversial. Low-stage, low-grade tumors can generally be cured by surgery alone. Postoperative radiotherapy improves locoregional control in all other tumor stages and grades. Currently, the diagnostic and therapeutic approach to parotid cancer offer few options for a class of neoplasms that has many subtypes each with a unique molecular background and variable clinical behavior. Nonetheless, this approach results in a satisfactory locoregional cancer control, making distant metastasis the most frequent cause for treatment failure. At present, systemic treatment for distant failure is disappointing, although recent progress in molecular biology has suggested that adding targeted therapy should achieve tumor response or stabilization. Although disease control remains variable, the prognosis of individual patients can be increasingly accurately predicted by multivariate analysis. PMID- 21618325 TI - Tobacco carcinogen metabolites and DNA adducts as biomarkers in head and neck cancer: potential screening tools and prognostic indicators. AB - Head and neck squamous cell carcinoma (HNSCC) is one of many cancers that are strongly associated with tobacco use. Whereas HNSCC is often seen in tobacco users, many tobacco users do not develop carcinoma; and the differences between smokers with and without HNSCC are poorly studied to date. Some smokers may be inherently more susceptible to developing carcinoma due to patterns of tobacco use, innate metabolism of carcinogens, or altered excretion. Identifying those smokers at greatest risk for HNSCC would have great benefit through targeted smoking cessation efforts and enhanced surveillance. One approach to better understand the extent of exposure to, and metabolism of, tobacco carcinogens is through the use of tobacco-specific metabolites. Tobacco-specific metabolites can identify patterns of dose, exposure, and metabolism, and perhaps ultimately characterize the important differences between smokers who develop HNSCC and smokers who do not. PMID- 21618327 TI - The DETECT adherence score--structure and psychometric exploration of a novel approach to measure adherence to drug and non-drug interventions in primary care. AB - There is substantial evidence that patient compliance or rather adherence to medical measures and recommendations for lifestyle changes can pivotally influence the prognosis of the patients or disease progression. However, the scientific evaluation and the statistical analysis of "patient adherence" are extremely difficult due to the fact that the construct of "adherence" is complex and comprised of many layers, and varies greatly in different disease groups. With this paper, we describe the development and structure of this novel assessment tool that takes past and prospective information on different facets of drug and behavioural adherence into account, expected to result in considerably improved prediction of future cardiovascular risk. We suggest a simple scoring scheme and explore the psychometric properties and the higher order factorial structure. In this exploratory study the "Diabetes Cardiovascular Risk Evaluation Targets and Essential Data for Commitment of Treatment" (DETECT) adherence score revealed good psychometric properties in terms of internal consistency and factorial structure, suggesting that its further exploration in terms of external validity is promising. Findings also underline that it is useful and informative to cover within one score both, pharmacologic and non pharmacologic interventions in primary care. Our combination in this respect is unique, as most studies conducted on this subject so far aimed at assessing solely drug adherence or behavioural adherence. PMID- 21618328 TI - Bone metastases: evaluation of acuity of lesions using dynamic gadolinium-chelate enhancement, preliminary results. AB - PURPOSE: To evaluate whether enhancement on serial dynamic gadolinium-enhanced abdominal-pelvic MR imaging (DCE-MRI) can determine the acuity of bone metastases. MATERIALS AND METHODS: Twenty consecutive patients who underwent abdominal-pelvic DCE-MRI for evaluation/staging of a proven cancer and had bone metastases were included. Two radiologists analyzed in consensus 59 DCE-MRIs of these patients. Region of interest measurements were performed in up to three lesions on noncontrast T1-weighted, serial hepatic arterial dominant phase (HADP), early hepatic venous phase (EHVF), and interstitial phase (IP) postgadolinium images, and the percentage enhancement of 134 lesions was calculated. The coordinator separately and retrospectively sorted the lesions into three groups based on the imaging and clinical information: acute/active, subacute, and chronic metastases. RESULTS: The mean percentage enhancement of the bone metastases classified as acute/active, subacute, and chronic in the HADP, EHVP and IP were respectively (%): 134, 107, 99; 87, 86, 87; and 39, 65, 73. In the HADP, acute/active lesions enhanced significantly more than both subacute (1.53-fold) and chronic (3.4-fold) lesions (P < 0.01). Time intensity curves were significantly different between these three entities as well. CONCLUSION: The enhancement on arterial phase images and the time-intensity curves were different for acute/active, subacute, and chronic bone metastases. PMID- 21618329 TI - Quantitative and reproducibility study of four tractography algorithms used in clinical routine. AB - PURPOSE: To evaluate fiber tracking strategy in terms of acquisition schemes in conjunction with four algorithms used in clinical routine, we studied one of the major tracts, anatomically well known, and which should be preserved as much as possible during neurosurgery: the corticospinal tract. MATERIALS AND METHODS: Two identical exams, composed of three DTI acquisition schemes (6, 15, and 32 gradient directions), were performed on 12 healthy subjects during two different sessions. For each subject, intra-operator, and inter-exam reproducibility was quantitatively calculated from different fiber tracking algorithms: three deterministic and a probabilistic one. Inter-exam reproducibility was evaluated comparing fiber tracking results from the repetition of the same acquisition one month apart and variation of the fiber density distribution percentile. RESULTS: For each fiber tracking algorithm, the best reproducibility result is obtained in case of 50% of fiber density and for the number of directions equal to 32. The reproducibility is improved using the probabilistic algorithm. CONCLUSION: This study highlights increased reliability of reproducibility results based on the number of directions used during the acquisition. The method of tractography used and the choice of adequate density fiber tract greatly improve the results. PMID- 21618330 TI - Robust mapping of the myelin water fraction in the presence of noise: synergic combination of anisotropic diffusion filter and spatially regularized nonnegative least squares algorithm. AB - PURPOSE: To improve the mapping of myelin water fraction (MWF) despite the presence of measurement noise, and to increase the visibility of fine structures in MWF maps. MATERIALS AND METHODS: An anisotropic diffusion filter (ADF) was effectively combined with a spatially regularized nonnegative least squares algorithm (srNNLS) for robust MWF mapping. Synthetic data simulations were performed to assess the effectiveness of this new method. Experimental measurements of signal decay curves were obtained and MWF maps were estimated using the new method and compared with maps estimated using other methods. RESULTS: MWF mapping was substantially improved in both simulations and experimental data when ADF was combined with the srNNLS algorithm. MWF variability decreased with the use of the proposed method, which in turn resulted in increased visibility of small focal lesions and structures in the MWF maps. CONCLUSION: This study demonstrates that the benefits of ADF and srNNLS algorithms can be effectively combined in a synergic way for robust mapping of MWF in the presence of noise. Substantial improvements to MWF mapping can be made using the proposed method. PMID- 21618331 TI - Transmural heterogeneity of left ventricular myocardium remodeling in postinfarct porcine model revealed by MR diffusion tensor imaging. AB - PURPOSE: To investigate the transmural heterogeneity of left ventricular myocardium structural remodeling. MATERIALS AND METHODS: Ex vivo diffusion tensor imaging (DTI) was performed in six adult porcine heart samples with apical septum infarction collected 13 weeks after permanent left anterior descending coronary artery ligation and six age-matched intact controls. Alterations in diffusion indices and myocardial fiber orientation, including fractional anisotropy (FA), mean apparent diffusion coefficient (mean ADC), axial diffusivity (lambda(?) ), radial diffusivity (lambda(?) ), and fiber helix angle were investigated at five transmural zones across myocardium wall in regions adjacent and remote to the infarct. RESULTS: In both adjacent and remote regions of infarcted hearts, FA showed no significant alteration across transmural zones compared to controls. However, mean ADC, lambda(?) , and lambda(?) exhibited significant decreases at endocardium zones but not epicardium zones. Moreover, myocardial fiber helix angle shifted towards left-handed orientation at all transmural zones, especially in regions adjacent to the infarct, becoming more aligned with the epicardium fiber orientation. CONCLUSION: These experimental DTI findings indicate that the endocardium was more vulnerable to infarction, leading to more pronounced microstructural changes during remodeling. The current DTI approach reveals additional information in delineating postinfarct remodeling process, which may provide insights into cardiac mechanics and clinical assessment of cardiac diseases. PMID- 21618332 TI - MR perfusion-weighted imaging may help in differentiating between nonenhancing gliomas and nonneoplastic lesions in the cervicomedullary junction. AB - PURPOSE: To evaluate the ability of dynamic susceptibility-weighted contrast enhanced magnetic resonance (MR) perfusion imaging (DSC-PWI) in distinguishing between nonenhancing gliomas and nonenhancing, nonneoplastic lesions in the cervicomedullary junction region. MATERIALS AND METHODS: This retrospective study involved eight patients with nonenhancing gliomas in the medulla oblongata and eight patients with nonenhancing nonneoplastic lesions. The relative cerebral blood volume (rCBV) ratios, peak heights, and percentage of signal intensity recovery derived from time-signal intensity curves of these nonenhancing lesions were compared. RESULTS: The mean peak height of nonenhancing gliomas was significantly higher than the value of their reference regions of interest (ROIs). In contrast, mean peak height of nonneoplastic lesions was significantly lower than their reference ROIs. The mean peak height and mean maximal rCBV ratio of nonenhancing gliomas were significantly higher than those of nonenhancing, nonneoplastic lesions (P<0.05). There was no significant difference with regard to percentage of signal intensity recovery between the two groups. CONCLUSION: DSC-PWI could be a useful adjuvant tool to differentiate between nonenhancing gliomas and nonenhancing, nonneoplastic lesions in the cervicomedullary junction region. PMID- 21618333 TI - Diagnostic value of whole-body magnetic resonance imaging for bone metastases: a systematic review and meta-analysis. AB - PURPOSE: To assess the overall diagnostic accuracy of whole-body magnetic resonance imaging (WB-MRI) in detecting bone metastases with a meta-analysis. MATERIALS AND METHODS: The MEDLINE, EMBASE, Cancerlit, and Cochrane Library databases were searched from January 1995 to September 2010 for studies evaluating the accuracy of WB-MRI in detecting bone metastases. Histopathologic analysis and/or close clinical and imaging follow-up for at least 6 months was assessed. Meta-analysis methods were used to pool sensitivity and specificity and to construct summary receiver-operating characteristics. RESULTS: A total of 11 studies with 495 patients who fulfilled all of the inclusion criteria were considered for the analysis. No publication bias was found. WB-MRI had a pooled sensitivity of 0.899 (95% confidence interval [CI], 0.845-0.939) and a pooled specificity of 0.918 (95% CI, 0.882-0.946). The subgroup without diffusion weighted imaging (DWI) positive results had higher pooled specificity 0.961 (95% CI, 0.922-0.984) than the subgroup with DWI (P < 0.05). CONCLUSION: WB-MRI was an accurate, cost-effective tool in detecting bone metastases. WB-MRI without DWI may improve the specificity of detecting bone metastases. DWI seems to be a sensitive but rather unspecific modality for the detection of bone metastatic disease. High-quality prospective studies regarding WB-MRI in detecting bone metastases still need to be conducted. PMID- 21618334 TI - Comparison of a 32-channel with a 12-channel head coil: are there relevant improvements for functional imaging? AB - PURPOSE: To evaluate the suitability of a 12- or 32-channel head coil and of a prescan normalization filter for functional magnetic resonance imaging (fMRI) studies at different brain regions. MATERIALS AND METHODS: fMRI was obtained from 36 volunteers executing a visually instructed motor paradigm using a 12-channel head matrix coil and a 32-channel phased-array head coil with and without prescan normalization filtering at 3 T. The time-course signal-to-noise ratio (tSNR) and the magnitude of functional activation (beta-value, t-value, percent signal change) were statistically compared between experimental conditions for the contralateral primary motor and visual cortex, contralateral thalamus, and ipsilateral anterior cerebellar hemispheres. RESULTS: tSNR was higher overall measuring with the 32-channel array and with prescan normalization. Without filtering, the 32-channel array delivered higher functional activation magnitudes for the visual cortex, whereas the 12-channel array seemed superior in this respect in thalamus and cerebellum. Filtering did not considerably affect the fMRI-activation magnitude detected from the 12-channel coil; its application favored the 32-channel coil at the subcortical and cerebellar locations but disfavored it at the cortical ones. CONCLUSION: The 32-channel coil detected more fMRI-activation cortically but less subcortically than the 12-channel coil; prescan normalization improved activation parameters only at central brain structures. PMID- 21618335 TI - Direct magnetic resonance arthrography of the wrist with axial traction: a feasibility study to assess joint cartilage. AB - PURPOSE: To assess the impact of axial traction during acquisition of direct magnetic resonance (MR) arthrography of the wrist with regard to joint space width and amount of contrast material between the opposing cartilage surfaces. MATERIALS AND METHODS: Fifteen consecutive patients (12 male, mean age 38.1 years) were included in this Institutional Review Board-approved prospective study. Three-compartment wrist MR arthrographies were performed between October and December 2009 on a 3 T unit using a fat-suppressed T1-weighted isotropic high resolution volumetric interpolated breathhold examination (VIBE) sequence in the coronal plane, with and without axial traction (3 kg). Two radiologists measured radiocarpal (radioscaphoid, radiolunate) and midcarpal (lunocapitate, hamatolunate) joint space widths, with and without traction, and assessed the amount of contrast material between the opposing cartilage surfaces using a three point scale: 0 = absence, 1 = partial, 2 = complete. RESULTS: With traction, joint space width increased significantly at the radioscaphoid (Delta = 0.78 mm, P < 0.01), radiolunate (Delta = 0.18 mm, P < 0.01), and lunocapitate (Delta = 0.45 mm, P < 0.01) spaces, and both observers detected significantly more contrast material between the cartilage surfaces. At the hamatolunate space, the differences in joint space width (Delta = 0.14 mm, P = 0.54) and amount of contrast material were not significant. CONCLUSION: Direct wrist MR arthrography with axial traction of 3 kg increases joint space width at the radiocarpal and lunocapitate spaces, and prompts better coverage of the articular cartilage by the contrast material. PMID- 21618336 TI - An analysis algorithm for accurate determination of articular cartilage thickness of hip joint from MR images. AB - PURPOSE: To test the accuracy of the most widely used technique based on edge detection for thickness measurement of the hip joint cartilage in MR images, and to improve the measurement accuracy by developing a new measurement method based on a model of the MRI process. MATERIALS AND METHODS: MRI was performed in 3 normal cadaver hips, 25 hips of normal volunteers, and 25 hips of patients with osteoarthritis. In general, thickness was defined as the distance between the two sides of the hip cartilage along the normal directions of the cartilage surfaces. In this article this is referred to as the "edge detection method." A theoretical simulation analysis revealed that the edge detection method considerably underestimated the cartilage thickness of the hip joint. A new measurement method based on a model of the MR imaging process was accordingly proposed for correcting the measurement errors. RESULTS: In the experiment using the cadaver hips, anatomical measurement of cartilage thickness was used as reference standard. For measurements at 35 sites, the proposed model-based method gave results similar to those presented from anatomic section, while the edge detection method gave underestimation compared with the anatomic thickness. The underestimation biases for the edge detection method were consistent with the biases predicted by theoretical simulation. In the experiment using the hips of volunteers and patients, the edge detection result was an underestimation compared with the result generated by using the model-based method. CONCLUSION: The edge detection method underestimated the hip cartilage thickness in MR images. The proposed model-based method was more accurate than the edge detection method for thickness measurement of the hip cartilage. PMID- 21618337 TI - New respiratory gating technique for whole heart cine imaging: integration of a navigator slice in steady state free precession sequences. AB - PURPOSE: To evaluate the performance of a slice navigator sequence integrated into a b-SSFP sequence for obtaining real time respiratory self-gated whole heart cine imaging. MATERIALS AND METHODS: In this work, we present a novel and robust approach for respiratory motion detection by integrating a slice navigator sequence into a balanced steady state free precession (b-SSFP) sequence, while maintaining the steady state. The slice navigator sequence is integrated into consecutive repetition times (TRs) of a b-SSFP sequence to excite and read out a navigator slice. We performed several phantom experiments to test the performance of the slice navigator sequence. Additionally, the method was evaluated in five volunteers and compared with breathing signals obtained from conventional pencil beam navigator sequence. Finally, the navigator slice was used to obtain whole heart MR cine images. RESULTS: The breathing signals detected by the proposed method showed an excellent agreement with those obtained from pencil beam navigators. Moreover, the technique was capable of removing respiratory motion artifacts with minimal distortion of the steady state. Image quality comparison showed a statistical significant improvement from a quality score of 2.1 obtained by the nonrespiratory gated images, compared to a quality score of 3.4 obtained by the respiratory gated images. CONCLUSION: This novel method represents a robust approach to estimate breathing motion during SSFP imaging. The technique was successfully applied to acquire whole heart artifact-free cine images. PMID- 21618338 TI - Skull-stripping method for brain MRI using a 3D level set with a speedup operator. AB - PURPOSE: To extract the brain region from brain magnetic resonance (MR) images using a fast 3D level set method and a refinement process. MATERIALS AND METHODS: The proposed method introduces a speedup operator to the conventional 3D level set method in order to accelerate the level set evolution. While the processing time for brain extraction is reduced by the speedup operator, the accuracy of brain extraction is also improved by adopting a refinement process. RESULTS: The speedup operator yielded a 75% reduction in the total iteration numbers for the synthesized volume. The proposed method was applied to several datasets and compared with previous methods, ie, BrainVisa, BET, and FreeSurfer. The proposed method provided a Jaccard index of 0.971 +/- 0.0114 for the BrainWeb dataset, 0.864 +/- 0.035 for the IBSR dataset, and 0.9414 +/- 0.0517 for a self-produced dataset acquired with a 3T MRI system. CONCLUSION: Utilizing a speedup operator, the proposed method reduced the evolution time. Robust and accurate results for various datasets were obtained in experiments. PMID- 21618339 TI - The central role of pharmacoepidemiology in comparative effectiveness research education: critical next steps. PMID- 21618340 TI - Risk of hospitalization for upper gastrointestinal adverse events associated with nonsteroidal anti-inflammatory drugs: a nationwide case-crossover study in Taiwan. AB - PURPOSE: This study aimed to evaluate the risks of upper gastrointestinal (GI) adverse events across a variety of oral and parenteral coxibs and nonselective nonsteroidal anti-inflammatory drugs (nsNSAIDs) in the general population of Taiwan. METHODS: In a case-crossover study, all patients aged >=20 years who were hospitalized for upper GI adverse events (peptic ulcer and bleeding; gastritis and duodenitis) in 2006 were identified using the International Classification of Diseases, 9th Revision, Clinical Modification diagnosis codes from inpatient claims from the Taiwan National Health Insurance Database. For each patient, the case period was defined as 1-30 days and the control period as 31-60 days before the date of hospitalization. Outpatient pharmacy prescription database was searched for individual NSAID use during the case and control periods. A conditional logistic regression model was applied, and adjusted self-matched odds ratios (OR) and their 95% confidence intervals (95%CI) were reported. RESULTS: A total of 40,635 patients hospitalized for upper GI adverse events were included. The adjusted OR was 1.52 (95%CI: 1.27-1.82) for celecoxib and 2.56 (95%CI: 2.44 2.69) for oral nsNSAIDs. The ORs were above 2 for oral piroxicam, diclofenac, ketorolac, ketoprofen, acemetacin, and naproxen and were around 1.5 for tiaprofenic acid, indomethacin, mefenamic acid, and ibuprofen. Higher risks were evident for parenteral NSAIDs, in particular ketorolac with an OR of 5.76 (95%CI: 5.14-6.44). CONCLUSION: Use of celecoxib and all nsNSAIDs studied was associated with a greater risk of upper GI toxicity as compared with nonuse. Parenteral NSAIDs posed a higher risk, but celecoxib, ibuprofen, and mefenamic acid posed a lower risk than other NSAIDs. PMID- 21618341 TI - Monozygotic twins discordant for neurofibromatosis type 1 due to a postzygotic NF1 gene mutation. AB - The analysis of monozygotic twins (MZ) concordant for neurofibromatosis type 1 (NF1) has indicated that genetic factors exert a major influence on the clinical variability (e.g. the number of cafe-au-lait spots and/or neurofibromas) evident in this disease. Here, we report on a pair of monozygotic, dichorionic twins who are phenotypically discordant with respect to NF1. Whereas DNA sequence analysis indicated somatic mosaicism for the NF1 nonsense mutation, c.4108C>T (p.Q1370X), in the affected twin II/1, this lesion was apparently absent in his unaffected brother. The observation of heterozygosity for flanking SNP and microsatellite markers rendered it most unlikely that the observed mosaicism with normal cells was due to mutation reversion brought about either by gene conversion or mitotic recombination. Instead, we conclude that the twinning event, which would have taken place within three days post-fertilization, must have preceded the c.4108C>T mutation which is therefore predicted to have occurred during the blastocyst stage, leading to somatic mosaicism with normal cells lacking the mutation. This is the first reported case of monozygotic twins discordant for NF1 in whom mosaicism for a postzygotic NF1 gene mutation has been observed in the affected but not the unaffected twin. PMID- 21618343 TI - Germline mutations in the PALB2 gene are population specific and occur with low frequencies in familial breast cancer. AB - The Partner and Localizer of BRCA2 (PALB2) protein has been linked to Fanconi anemia and breast cancer predisposition. Here we present data of a comprehensive mutation screening of the PALB2 gene in 818 familial cases of breast cancer from Germany. By analyzing the entire coding region of PALB2, we found seven truncating mutations (six of them novel) in families tested negative for BRCA1/2 mutations. In addition, two novel potentially disease causing missense mutations were found. Remarkably, only one mutation reported previously in other populations, was also identified in the German population. No PALB2 mutation carriers were identified in 450 unaffected controls. Thus, our observations indicate a low prevalence of deleterious PALB2 mutations and a specific mutation profile within the German population. As PALB2-deficient tumors were shown to be sensitive to Poly(ADP-ribose) Polymerase (PARP) inhibitors, our study has implications for newly developed, favorable treatment options in familial breast cancer. PMID- 21618342 TI - Analysis of the disintegrin-metalloproteinases family reveals ADAM29 and ADAM7 are often mutated in melanoma. AB - We performed a mutational analysis of the 19 disintegrin-metalloproteinases (ADAMs) genes in human cutaneous metastatic melanoma and identified eight to be somatically mutated in 79 samples, affecting 34% of the melanoma tumors analyzed. Functional analysis of the two frequently mutated ADAM genes, ADAM29 and ADAM7 demonstrated that the mutations affect adhesion of melanoma cells to specific extracellular matrix proteins and in some cases increase their migration ability. This suggests that mutated ADAM genes could play a role in melanoma progression. PMID- 21618344 TI - Characterization of two mutations in the SPTLC1 subunit of serine palmitoyltransferase associated with hereditary sensory and autonomic neuropathy type I. AB - Hereditary sensory and autonomic neuropathy type I (HSAN-I) is an axonal peripheral neuropathy leading to progressive distal sensory loss and severe ulcerations. Mutations in SPTLC1 and SPTLC2, encoding the two subunits of serine palmitoyltransferase (SPT), the enzyme catalyzing the first and rate-limiting step in the de novo synthesis of sphingolipids, have been reported to cause HSAN I. Here, we demonstrate that the SPTLC1 mutations p.S331F and p.A352V result in a reduction of SPT activity in vitro and are associated with increased levels of the deoxysphingoid bases 1-deoxy-sphinganine and 1-deoxymethyl-sphinganine in patients' plasma samples. Stably expressing p.S331F-SPTLC1 HEK293T cell lines likewise show accumulation of deoxysphingoid bases, but this accumulation is not observed in HEK293T cells overexpressing p.A352V-SPTLC1. These results confirm that the increased formation of deoxysphingoid bases is a key feature for HSAN-I as it is associated with all pathogenic SPTLC1 and SPTLC2 mutations reported so far, but also warrant for caution in the interpretation of in vitro data. PMID- 21618345 TI - miRvar: A comprehensive database for genomic variations in microRNAs. AB - microRNAs are a recently discovered and well studied class of small noncoding functional RNAs. The regulatory role of microRNAs (miRNAs) has been well studied in a wide variety of biological processes but there have been no systematic effort to understand and analyze the genetic variations in miRNA loci and study its functional consequences. We have comprehensively curated genetic variations in miRNA loci in the human genome and established a computational pipeline to assess potential functional consequences of these variants along with methods for systematic curation and reporting of variations in these loci. The data is made available on the Leiden Open (source) Variation Database (LOVD) platform at http://genome.igib.res.in/mirlovd to provide ease of aggregation and analysis and is open for community curation efforts. PMID- 21618346 TI - Next generation sequencing of pooled samples reveals new SNRNP200 mutations associated with retinitis pigmentosa. AB - The gene SNRNP200 is composed of 45 exons and encodes a protein essential for pre mRNA splicing, the 200 kDa helicase hBrr2. Two mutations in SNRNP200 have recently been associated with autosomal dominant retinitis pigmentosa (adRP), a retinal degenerative disease, in two families from China. In this work we analyzed the entire 35-Kb SNRNP200 genomic region in a cohort of 96 unrelated North American patients with adRP. To complete this large-scale sequencing project, we performed ultra high-throughput sequencing of pooled, untagged PCR products. We then validated the detected DNA changes by Sanger sequencing of individual samples from this cohort and from an additional one of 95 patients. One of the two previously known mutations (p.S1087L) was identified in 3 patients, while 4 new missense changes (p.R681C, p.R681H, p.V683L, p.Y689C) affecting highly conserved codons were identified in 6 unrelated individuals, indicating that the prevalence of SNRNP200-associated adRP is relatively high. We also took advantage of this research to evaluate the pool-and-sequence method, especially with respect to the generation of false positive and negative results. We conclude that, although this strategy can be adopted for rapid discovery of new disease-associated variants, it still requires extensive validation to be used in routine DNA screenings. PMID- 21618347 TI - Classical investigation of incomplete collagen C-propeptide processing reveals a distinctive high bone mass OI phenotype. PMID- 21618348 TI - Comparison of programs for in silico assessment of missense substitutions. PMID- 21618349 TI - Functional characterization of naturally occurring genetic variants in the human TLR1-2-6 gene family. AB - Toll-like receptors (TLRs) are considered an essential component of the innate immune system, initiating inflammatory responses following infection of the host. Humans have 10 functional TLRs, differing in their subcellular distributions and the microbial agonists they sense. The phylogenetically conserved TLR1-2-6 family is unique in that TLR1 and TLR6 form heterodimers with TLR2 to mediate signalling in response to agonists. Epidemiological genetic studies have identified several TLR variants that appear to influence susceptibility to infectious diseases, but the functional consequences of which remain largely unknown. Here, we assessed the functional impact of the TLR1-2-6 variants with altered amino acid sequences segregating naturally in the human population. We used an NF-kappaB reporter assay in TLR-transfected human embryonic kidney 293T cells stimulated with the corresponding TLR agonists. We found that among the 41 naturally occurring variants with amino acid alterations identified in the TLR1-2-6 family, 14 of them (five TLR1, four TLR2, and five TLR6 variants) displayed marked impairment of NF-kappaB activation. Most of these variants are present at very low population frequencies and are population-specific. These observations suggest that rare, nonsynonymous TLR mutations are likely to have deleterious effects on immune responses and may therefore contribute to complex susceptibility to infection at the population level. PMID- 21618350 TI - Relative role of methylator and tumor suppressor pathways in ulcerative colitis associated colon cancer. AB - BACKGROUND: Chronic ulcerative colitis (UC) is associated with an increased colorectal cancer risk which may be secondary to repetitive mucosal injury. Both epigenetic methylation and the classic adenoma-to-carcinoma sequence have been implicated in this malignant transformation, but the underlying molecular mechanisms remain poorly defined. This study compares the molecular characteristics of colitis-associated and common colorectal cancers. METHODS: Nineteen patients with colorectal adenocarcinomas arising within UC were matched for age and cancer site with 54 patients with sporadic adenocarcinomas. Tumor tissue was examined for BRAF mutations, CpG island methylator phenotype (CIMP), and MLH1 promoter methylation. Mutations of KRAS and p53 were assessed by sequencing. RESULTS: Patient demographics were similar for the two groups. CIMP was observed in 22% of sporadic colorectal cancers and in 5% of UC cancers (P = 0.162). Rates of BRAF mutation (4% vs 5%, P = 1.0), MLH1 methylation (9% versus 5%, P = 0.682), and KRAS mutations (24% versus 32%, P = 0.552) were similar between the groups. However, colitis-associated colorectal cancers were more likely to have a p53 mutation compared to sporadic adenocarcinomas (95% versus 53%, P = 0.001). The dominant mutation for colitis-associated cancers was a mutation in codon 4, representing half of the mutations. Furthermore, colitis associated cancers had a higher rate of mutation in codon 8 (48% versus 6%, P < 0.001) than sporadic counterparts. CONCLUSIONS: Unlike other inflammatory gastrointestinal cancers, colitis-associated colorectal cancers do not preferentially arise via a methylator pathway when compared to sporadic colorectal cancers. Chromosomal instability remains an important etiology, but with a unique p53 frequency and mutation pattern. PMID- 21618351 TI - Expression of HNF4gamma is downregulated in patients with active ulcerative colitis (UC) compared to UC patients in remission and healthy controls. PMID- 21618352 TI - Clinical outcomes and prognostic factors for thiopurine maintenance therapy in patients with intestinal Behcet's disease. AB - BACKGROUND: To date, there have been no studies focusing on the efficacy of thiopurine therapy in intestinal Behcet's disease (BD). We conducted this study to investigate clinical outcomes and predictors of clinical relapse in intestinal BD patients receiving thiopurine maintenance therapy. METHODS: We reviewed the medical records of all patients with intestinal BD who received thiopurine therapy in a single tertiary academic medical center between March 1986 and October 2010. The cumulative probabilities of clinical relapse after remission were calculated using the Kaplan-Meier method. Predictors of clinical relapse were identified by univariate analysis using the log-rank test and by multivariate analysis using Cox proportional hazards regression models. RESULTS: Of a total of 272 patients with intestinal BD, 67 (24.6%) received their first course of thiopurine therapy at our center. Thirty-nine (58.2%) of the 67 patients constantly received thiopurines for maintaining medically or surgically induced remission. The cumulative relapse rates at 1 year, 2 years, 3 years, and 5 years after remission were 5.8%, 28.7%, 43.7%, and 51.7%, respectively. On multivariate analysis, a younger age (<25 years) at diagnosis and a lower hemoglobin level (<11 g/dL) were independent predictive factors for relapse in intestinal BD patients receiving thiopurine maintenance therapy. CONCLUSIONS: Thiopurine therapy showed a relatively good effect for maintenance of remission in intestinal BD patients. However, a younger age at diagnosis and a lower hemoglobin level were associated with a poor response to thiopurines, necessitating early adoption of effective alternative therapeutic options in these risk groups. PMID- 21618353 TI - Long-term durability of response to adalimumab in Crohn's disease. AB - BACKGROUND: Adalimumab is an effective treatment for Crohn's disease (CD), but may also be associated with loss of response. Few reports provide insight into the durability of treatment of CD with adalimumab for periods longer than 12 months in clinical practice. AIMS: To evaluate the long-term durability of adalimumab maintenance treatment and to identify predictive factors associated with loss of response. METHODS: CD patients who initially responded to adalimumab were evaluated in a historical cohort study. Maintenance of long-term response was estimated using Kaplan-Meier analysis. Cox regression analysis was performed to identify potential predictive factors for loss of efficacy. RESULTS: In all, 380 CD patients were included (mean age, 38 years; 52% female). Of these, 43% had ileocolic CD, 50% inflammatory CD, and 41% perianal CD. Median follow-up with adalimumab was 8 months (range, 4-75 months). The annual risk of loss of response to adalimumab was 18% per patient-year of follow-up. Twenty-eight percent of patients were anti-TNF-naive and 72% anti-TNF-experienced. The loss of efficacy was 8% per patient-year of follow-up in the anti-TNF-naive patients and 22% in the anti-TNF-experienced group (P < 0.01). In the multivariate analysis, the presence of extraintestinal manifestations (hazard ratio [HR] = 1.7; 95% confidence interval [CI] = 1.02-2.9) and previous experience with other anti-TNF agents (HR = 2.5,95% CI = 1.2-5.3) were associated with higher risk of loss of efficacy. CONCLUSIONS: A relevant proportion of CD patients on long-term adalimumab lost response. The risk of loss of response was higher (more than 2 fold) in anti-TNF-experienced than in anti-TNF-naive patients (22% vs. 8% per patient-year of treatment). Having extraintestinal manifestations seems to increase the risk of loss of efficacy. PMID- 21618354 TI - Fructo-oligosaccharides as therapeutics for active Crohn's disease: adequate power and no effect. PMID- 21618355 TI - Multifocal pouch body adenocarcinoma following ileal pouch-anal anastomosis (IPAA) for ulcerative colitis. PMID- 21618356 TI - Dextran sodium sulfate inhibition of real-time polymerase chain reaction amplification: a poly-A purification solution. AB - BACKGROUND: Dextran sulfate sodium (DSS) induces experimental colitis and promotes colitis-associated cancer in rodents. Here we document potent inhibition of real-time quantitative polymerase chain reaction (qPCR) using cDNA from DSS exposed mouse tissues, which complicates gene expression analysis. METHODS: We characterize DSS inhibition of qPCR in-vitro and in a wide array of murine tissues following ingestion of DSS. We examine different approaches to RNA purification prior to cDNA synthesis in order to optimize real-time polymerase chain reaction amplification and gene expression analysis. RESULTS: DSS inhibits qPCR amplification of cDNA between 1 and 10 nM. Orally administered DSS interferes with qPCR amplification of cDNA derived from multiple tissues. Poly-A purification of DSS-exposed RNA allows reliable and cost-effective gene expression analysis in DSS-exposed tissue. CONCLUSIONS: DSS is a potent inhibitor of real-time qPCR amplification and interferes with tissue-specific gene expression analysis in DSS-exposed mice. Poly-A purification of tissue-derived RNA results in reliable and cost-effective gene expression analysis in DSS exposed mice. PMID- 21618357 TI - CESAME substudy on thiopurines in pregnancy: is this the baby we were waiting for? PMID- 21618358 TI - Information needs and preferences in IBD (IBD self-management: the AGA guide to Crohn's disease and ulcerative colitis). PMID- 21618359 TI - B cells exposed to enterobacterial components suppress development of experimental colitis. AB - BACKGROUND: B cells positively contribute to immunity by antigen presentation to CD4(+) T cells, cytokine production, and differentiation into antibody secreting plasma cells. Accumulating evidence implies that B cells also possess immunoregulatory functions closely linked to their capability of IL-10 secretion. METHODS: Colitis development was followed in CD4(+) CD25(-) T cell transplanted SCID mice co-transferred with B cells exposed to an enterobacterial extract (ebx B cells). B and T cell cytokine expression was measured by flow cytometry and enzyme-linked immunosorbent assay (ELISA). RESULTS: We demonstrate that splenic B cells exposed to ebx produce large amounts of IL-10 in vitro and express CD1d and CD5 previously known to be associated with regulatory B cells. In SCID mice transplanted with colitogenic CD4(+) CD25(-) T cells, co-transfer of ebx-B cells significantly suppressed development of colitis. Suppression was dependent on B cell-derived IL-10, as co-transfer of IL-10 knockout ebx-B cells failed to suppress colitis. Ebx-B cell-mediated suppression of colitis was associated with a decrease in interferon gamma (IFN-gamma)-producing T(H) 1 cells and increased frequencies of Foxp3-expressing T cells. CONCLUSIONS: These data demonstrate that splenic B cells exposed to enterobacterial components acquire immunosuppressive functions by which they can suppress development of experimental T cell-mediated colitis in an IL-10-dependent way. PMID- 21618360 TI - Reciprocal regulation of the survival and apoptosis of Th17 and Th1 cells in the colon. AB - BACKGROUND: The immediate early response gene X-1 (IEX-1) is a stress-inducible gene involved in the regulation of cell growth, apoptosis and inflammation. METHODS: Acute colitis was induced by treatment of IEX-1 knockout (KO) and wild type (WT) control mice with dextran sulfate sodium (DSS), whereas chronic colitis was induced in Rag-/- mice by adoptive transfer of CD4(+) CD45RB(hi) T cells isolated from the two strains of mice. The diseases and responses of lamina propria lymphocytes were analyzed in the mice. RESULTS: IEX-1 KO mice produced IL 17 in the colon significantly greater than WT control mice following DSS treatment owing to better survival and differentiation of both IL-17-secreting gammadelta T cells and Th17 cells. The altered level of IL-17 production contributed critically to the reduced colon inflammation in IEX-1 KO mice, and administration of neutralizing anti-IL-17 antibody increased susceptibility of the animal to the disease. Strikingly, in contrast to the better survival of T cells producing IL-17, lack of IEX-1 enhanced apoptosis in proinflammatory T cells producing interferon gamma (IFN-gamma). Enhanced apoptosis in Th1 cells and better survival of Th17 cells may both result in a delayed onset of colitis in Rag-/- mice receiving pathogenic CD4(+) CD45RB(hi) T cells isolated from IEX-1 KO animals compared to those mice transferred with WT counterparts CONCLUSIONS: The present study demonstrates the refractoriness of IEX-1 knockout (KO) mice to DSS induced colitis and diminished pathogenesis of IEX-1-deficient CD4(+) CD45RB(hi) T cells. These data demonstrate that IEX-1 reciprocally regulates T-cell survival and apoptosis in a subset-dependent fashion. Inhibition of IEX-1 may thus offer novel strategies for colitis treatment by simultaneous induction of apoptosis in proinflammatory Th1 cells while promoting the survival and differentiation of a protective T-cell subset. PMID- 21618361 TI - Rectal neuroendocrine tumor in ulcerative colitis. PMID- 21618362 TI - Fecal bacteriotherapy for ulcerative colitis: patients are ready, are we? AB - BACKGROUND: Fecal bacteriotherapy (FB) has been proposed as a safe and effective alternative treatment for a number of gastrointestinal conditions including ulcerative colitis (UC). We performed a qualitative study to explore the attitudes and concerns of adult patients and parents of children with UC regarding FB as a potential treatment. METHODS: We conducted six focus groups for adult patients with UC and parents of children with UC or indeterminate colitis. Participants were asked about their perceptions of and interest in FB as a treatment for UC. Sessions were recorded, transcribed, and reviewed to identify domains, themes, and major concepts. RESULTS: The focus groups included 15 adult patients and seven parents of children with colitis. We identified five major domains pertaining to FB: impressions of treatment, benefits, risks, potential mechanisms, and social concerns. All but one participant expressed interest in FB and several wished it were already available. Participants compared FB to probiotics, felt it was "natural," easier than current therapies, and with donor screening would be safe. Although initial distaste and the "yuck factor" were uniformly mentioned, these concerns were outweighed by perceived benefits. CONCLUSION: This is the first study to examine important ethical and social issues surrounding FB as a treatment for UC. Given adequate supporting research, donor selection, and screening, adult patients and parents of children with UC will consider FB and are eager for it to become available. These findings have important implications for future microbiome-based treatments. PMID- 21618363 TI - Unfractionated or low molecular weight heparin for induction of remission in ulcerative colitis: a Cochrane inflammatory bowel disease and functional bowel disorders systematic review of randomized trials. AB - BACKGROUND: We aimed to systematically review the efficacy of unfractionated heparin (UFH) or low molecular weight heparins (LMWH) for remission induction in patients with ulcerative colitis (UC). METHODS: A literature search to April 2011 was performed to identify all randomized trials studying UFH or LMWH use in patients with UC. The Cochrane Risk of Bias Tool was used to assess study quality. RESULTS: LMWH administered subcutaneously showed no benefit over placebo for any outcome, including clinical remission, and clinical, endoscopic, or histological improvement. High-dose LMWH administered via an extended colon release tablet demonstrated benefit over placebo for clinical remission (odds ratio [OR] 2.73; 95% confidence interval [CI] 1.32-5.67; P = 0.007), clinical improvement (OR 2.99; 95% CI 1.30-6.87; P = 0.01), and endoscopic improvement (OR 2.25; 95% CI 1.01-5.01; P = 0.05) but not endoscopic remission or histologic improvement. LMWH was not beneficial when added to standard therapy for clinical remission, clinical improvement, endoscopic remission, or endoscopic improvement. One study examining UFH versus corticosteroids for the treatment of severe UC demonstrated the inferiority of UFH for clinical improvement. More patients assigned to UFH had rectal hemorrhage as an adverse event. CONCLUSIONS: LMWH administered by extended colon-release tablets may be effective for the treatment of active UC. This benefit needs to be confirmed by further randomized controlled studies. The same benefits were not seen when LMWH was administered subcutaneously at lower doses. There is no evidence to support the use of UFH for the treatment of active UC. PMID- 21618364 TI - Maternal inflammatory bowel disease and offspring body size: a prospective cohort study. AB - BACKGROUND: Maternal inflammatory bowel disease (IBD) may influence intrauterine growth and hence size at birth, but the consequences for offspring in later life remain uncertain. This study investigated the growth of children of mothers with Crohn's disease (CD) or ulcerative colitis (UC). METHODS: The Danish National Birth Cohort, comprising 40,640 mother-child pairs with 7-year follow-up of children's height and weight, were linked to the Danish National Disease Register, whereby 50 mothers with CD and 147 mothers with UC were identified. Associations were tested by regression analyses, taking several covariates into account. RESULTS: Children of mothers with IBD were significantly shorter at birth than children of unaffected mothers (adjusted; CD, difference in cm, -0.82; 95% confidence interval [CI], -1.39 to -0.25; UC, -0.41; 95% CI, -0.75 to -0.07) and they tended to be of lower birth weight (adjusted; CD, difference in grams, 119.7; 95% CI, -246.7 to 7.3; UC, -64.0; 95% CI, -138.7 to 10.7). However, during the first year of life children of IBD mothers reached similar body sizes as children of unaffected mothers. At the 7-year follow-up, girls, but not boys, of CD mothers tended to be overweight (adjusted odds ratio [OR], 2.47; 95% CI, 0.98 6.24) and had increased waist circumference (adjusted difference in cm, 3.48; 95% CI, 1.40-5.58) compared to the unaffected population. CONCLUSIONS: The present study confirms that maternal history of IBD leads to decreased birth size in offspring. Reassuringly, body size in children of IBD mothers approached body size in children of unaffected mothers during childhood. PMID- 21618365 TI - Transmission distortion in Crohn's disease risk gene ATG16L1 leads to sex difference in disease association. AB - BACKGROUND: Crohn's disease (CD), an inflammatory disease of the bowel, affects millions of people around the world. Evidence suggests that disease onset and pathogenesis differ between males and females. Yet no comprehensive efforts exist to assess the sex-specific genetic architecture of CD. METHODS: We used genotyping data from a cohort of 1748 CD cases and 2938 controls to investigate 71 meta-analysis-confirmed CD risk loci for sex differences in disease risk. We further validated the significant results in separate cohorts of 968 CD cases and 2809 controls, and performed a meta-analysis across datasets. RESULTS: The single nucleotide polymorphism (SNP) rs3792106 (C/T) in ATG16L1 showed a significant sex effect with P-value 6.9 * 10(-13) and allelic odds ratio 1.48 in females, and P value 0.013 and odds ratio 1.22 in males (odds ratio heterogeneity P-value 0.037). Surprisingly, the difference was found to arise from a discrepancy in allele frequencies between male and female controls (P-value 0.0045) rather than cases. We found similar results for this SNP in the separate validation datasets. Using 155 HapMap 3 trios, we detected significant maternal overtransmission of the T allele at rs3792106 (P-value 0.027). CONCLUSIONS: Our results indicate that different transmission patterns between sexes may sustain the disparate allele frequencies at rs3792106 in healthy populations, and furthermore that a virus risk variant mechanism implicated in CD alters the distribution in diseased patients. To our knowledge, this is the first report of sex-specific CD association in ATG16L1. The possible implications in CD and basic human biology present interesting areas for future investigation. PMID- 21618366 TI - Varicella zoster meningoradiculitis in Crohn's disease treated with 6 mercaptopurine. PMID- 21618367 TI - Controlled self-assembling of gadolinium nanoparticles as smart molecular magnetic resonance imaging contrast agents. PMID- 21618368 TI - One is enough: influencing polymer properties with a single chromophoric unit. PMID- 21618369 TI - Synthesis with perfect atom economy: generation of diazo ketones by 1,3-dipolar cycloaddition of nitrous oxide at cyclic alkynes under mild conditions. PMID- 21618370 TI - Cross-coupling reactions of organoboranes: an easy way to construct C-C bonds (Nobel Lecture). PMID- 21618371 TI - Electric control of magnetization and interplay between orbital ordering and ferroelectricity in a multiferroic metal-organic framework. PMID- 21618372 TI - Disrupting the reader of histone language. PMID- 21618373 TI - Coordination polymer nanostructures. PMID- 21618374 TI - Zwitterionic copolymerization: synthesis of cyclic gradient copolymers. PMID- 21618375 TI - A multidimensional sensing device for the discrimination of proteins based on manganese-doped ZnS quantum dots. AB - Lab-on-a-nanoparticle: the triple-channel optical properties of Mn-doped ZnS quantum dots (fluorescence, phosphorescence, and light scattering) are explored to develop a multidimensional sensing device for the discrimination of proteins in a lab-on-a-nanoparticle approach. PMID- 21618376 TI - Continuous in situ generation, separation, and reaction of diazomethane in a dual channel microreactor. PMID- 21618377 TI - Ion-induced transformation of magnetism in a bimetallic CuFe Prussian blue analogue. PMID- 21618379 TI - Slow photoelectron spectroscopy of delta-valerolactam and its dimer. AB - We studied the single-photon ionization of gas-phase delta-valerolactam (piperidin-2-one) and of its dimer using vacuum-ultraviolet (VUV) synchrotron radiation coupled to a velocity map imaging electron/ion coincidence spectrometer. The slow photoelectron spectrum (SPES) of the monomer is dominated by the vibrational transitions to the X state. Moreover, several weaker and complex bands are observed, corresponding to the population of the vibrational bands (pure or combination) of the electronically excited states of the cation arising from their mutual vibronic interactions. For the dimer, we measure a unique large band. These spectra are assigned with the help of theoretical calculations dealing with the equilibrium geometries, electronic-state patterns and evolutions, harmonic and anharmonic wavenumbers of the monomer and dimer, either neutral or positively charged. The state energies of the [delta valerolactam](+) cation in the X ground, A, B, C, excited electronic states, and of the [delta-valerolactam](2) (+) cation's lowest states are determined. After its formation, [delta-valerolactam](2) (+) is subject to intramolecular isomerization, H transfer and then unimolecular fragmentation processes. Close to the ionization thresholds, the photoionization of these molecules is found to be mainly dominated by a direct process whereas the indirect route (autoionization) contributes at higher energies. PMID- 21618380 TI - IR spectroscopy of isolated neutral and protonated adenine and 9-methyladenine. AB - IR spectroscopy is employed to study isolated adenine and its derivative 9 methyladenine in both their neutral and protonated forms. The IR spectra of neutral adenine and 9-methyladenine are measured in a molecular beam expansion via IR-UV ion-dip spectroscopy in the 525 to 1750 cm(-1) region. For adenine, UV excitation selects the 9H tautomer to give a conformer-selective IR spectrum. For 9-methyladenine, only one tautomer exists because of the methyl substitution at the N(9) position. The experimental spectra agree closely with spectra computed for these tautomers at the B3LYP/6-311++G(df,pd) level of theory. These spectra complement previous tautomer-specific IR spectra in the hydrogen stretching range. The 9H-adenine spectrum obtained is compared to a previously recorded FTIR spectrum of adenine at 280 degrees C, which shows close agreement, although the 7H tautomer cannot be excluded from contributing. Protonated adenine and 9 methyladenine are generated by electrospray ionization and studied via IR multiple-photon dissociation (IRMPD) spectroscopy. Comparison of the experimental spectra with computed spectra allows identification of the protonation site, which suggests that the 1-9 tautomer is the dominant contributor to the spectra. PMID- 21618381 TI - Teaching argumentation and scientific discourse using the ribosomal peptidyl transferase reaction. AB - Argumentation and discourse are two integral parts of scientific investigation that are often overlooked in undergraduate science education. To address this limitation, the story of peptide bond formation by the ribosome can be used to illustrate the importance of evidence, claims, arguments, and counterarguments in scientific discourse. With the determination of the first structure of the large ribosomal subunit bound to a transition state inhibitor came an initial hypothesis about the role of the ribosome in peptide bond formation. This initial hypothesis was based on a few central assumptions about the transition state mimic and acid-base catalysis by serine proteases. The initial proposed mechanism started a flurry of scientific discourse in experimental articles and commentaries that tested the validity of the initial proposed mechanism. Using this civil argumentation as a guide, class discussions, assignments, and a debate were designed that allow students to analyze and question the claims and evidence about the mechanism of peptide bond synthesis. In the end, students develop a sense of critical skepticism, and an understanding of scientific discourse, while learning about the current consensus mechanism for peptide bond synthesis. Biochemistry and Molecular Biology Education Vol. 39, No. 3, pp. 185-190, 2011. PMID- 21618382 TI - Assessment of genetics knowledge and skills in medical students: insight for a clinical neurogenetics curriculum. AB - The pace of discovery in biochemistry and genetics and its effect on clinical medicine places new curricular challenges in medical school education. We sought to evaluate students' understanding of neurogenetics and its clinical applications to design a pilot curriculum into the clinical neurology clerkship. We utilized a needs assessment and a written examination to evaluate the genetics knowledge of 81 third- and fourth-year medical students. The needs assessment surveyed students' self-perceptions of their own understanding of basic and clinically related genetic principles and clinical skills, as well as the most effective educational methods. Medical students reported more competence with basic science learned during the preclinical years than clinical concepts, and they demonstrated relatively low knowledge levels in clinical neurogenetics concepts on the examination, with an average of 29% correct on questions pertaining to genetic counseling compared with 82% correct with regard to inheritance patterns. Common, cross-specialty clinical skills were attained (e.g. internet search, family histories), while at least half of students reported minimal understanding or awareness of key genetics websites (e.g. OMIM) and indications for support group recommendations and genetics referrals. Teaching these more specific genetics skills and concepts needs to be emphasized in the clinical curriculum. PMID- 21618383 TI - A writing-intensive, methods-based laboratory course for undergraduates. AB - Engaging undergraduate students in designing and executing original research should not only be accompanied by technique training but also intentional instruction in the critical analysis and writing of scientific literature. The course described here takes a rigorous approach to scientific reading and writing using primary literature as the model while simultaneously integrating laboratory instruction on basic enzyme purification and characterization, followed by 6 weeks of laboratory dedicated to student-designed original research projects. In the preparation and execution of their original projects, students engage in analysis of the primary literature, proposal writing, peer review, manuscript preparation, and oral presentation. The result is a comprehensive and challenging course that teaches third- and fourth-year undergraduates what it means to "think and work like a scientist." PMID- 21618384 TI - A practical workshop for generating simple DNA fingerprints of plants. AB - Gel electrophoresis DNA fingerprints offer a graphical and visually appealing illumination of the similarities and differences between DNA sequences of different species and individuals. A polymerase chain reaction (PCR) and restriction digest protocol was designed to give high-school students the opportunity to generate simple fingerprints of plants thereby illustrating concepts and techniques in genetics and molecular biology. Three combinations of primers/restriction enzyme targeting chloroplast DNA were sufficient to generate patterns that enabled visual discrimination of plant species. The protocol was tested with a range of common fruit, vegetable, and herb plants that could be easily cultivated and handled in the laboratory. Toxic or hazardous materials such as ethidium bromide and liquid nitrogen were avoided. The protocol was validated as a university outreach workshop targeted at a group of up to 10 high school students. In a teaching laboratory, students sampled plants, setup the PCR reaction and restriction digest using microliter pipettes, and loaded the digested samples on an agarose gel. The workshop was structured as 2 * 2.5-hour sessions on separate days. The main challenges stemmed from the speed and accuracy of pipetting, especially at the gel loading stage. Feedback from students was largely positive, with the majority reporting that they had both enjoyed and learnt from the experience. PMID- 21618385 TI - An in silico DNA cloning experiment for the biochemistry laboratory. AB - This laboratory exercise introduces students to concepts in recombinant DNA technology while accommodating a major semester project in protein purification, structure, and function in a biochemistry laboratory for junior- and senior-level undergraduate students. It is also suitable for forensic science courses focused in DNA biology and advanced high school biology classes. Students begin by examining a plasmid map with the goal of identifying which restriction enzymes may be used to clone a piece of foreign DNA containing a gene of interest into the vector. From the National Center for Biotechnology Initiative website, students are instructed to retrieve a protein sequence and use Expasy's Reverse Translate program to reverse translate the protein to cDNA. Students then use Integrated DNA Technologies' OligoAnalyzer to predict the complementary DNA strand and obtain DNA recognition sequences for the desired restriction enzymes from New England Biolabs' website. Students add the appropriate DNA restriction sequences to the double-stranded foreign DNA for cloning into the plasmid and infecting Escherichia coli cells. Students are introduced to computational biology tools, molecular biology terminology and the process of DNA cloning in this valuable single session, in silico experiment. This project develops students' understanding of the cloning process as a whole and contrasts with other laboratory and internship experiences in which the students may be involved in only a piece of the cloning process/techniques. Students interested in pursuing postgraduate study and research or employment in an academic biochemistry or molecular biology laboratory or industry will benefit most from this experience. PMID- 21618386 TI - Differentiating biochemistry course laboratories based on student experience. AB - Content and emphases in undergraduate biochemistry courses can be readily tailored to accommodate the standards of the department in which they are housed, as well as the backgrounds of the students in the courses. A more challenging issue is how to construct laboratory experiences for a class with both chemistry majors, who usually have little or no experience with biochemical techniques and biology and biochemistry majors who do. This manuscript describes a strategy for differentiating biochemistry labs to meet the needs of students with differing backgrounds. PMID- 21618388 TI - The turn of the screw: an exercise in protein secondary structure. AB - An exercise using simple paper strips to illustrate protein helical and sheet secondary structures is presented. Drawing on the rich historical context of the use of physical models in protein biochemistry by early practitioners, in particular Linus Pauling, the purpose of this activity is to cultivate in students a hands-on, intuitive sense of protein secondary structure and to complement the common computer-based structural portrayals often used in teaching biochemistry. As students fold these paper strips into model secondary structures, they will better grasp how intramolecular hydrogen bonds form in the folding of a polypeptide into secondary structure, and how these hydrogen bonds direct the overall shape of helical and sheet structures, including the handedness of the alpha-helix and the difference between right- and the left handed twist. PMID- 21618390 TI - Commentary: How the internet is changing the way we think, read and remember. PMID- 21618391 TI - Proteopedia entry: citrate synthase. PMID- 21618392 TI - Websites of note. PMID- 21618393 TI - Bridging the educational research-teaching practice gap: curriculum development, part 2: becoming an agent of change. AB - Many faculty members in science departments are experiencing pressure to improve their courses, particularly with respect to the ways in which students are taught and assessed. The purpose of this article is to provide some insights and practical ideas on how curriculum change can be brought about-how motivated individuals can become agents of change. Change almost always elicits opposing and supporting forces, examples of which are given. Finally, we discuss examples of strategies to deal with these forces and highlight various factors that need to be considered when implementing such strategies, including the concepts of a zone of feasible innovation, the zone of tolerance, and the development of communities of practice. PMID- 21618394 TI - Endorsing alternative careers for PhDs. PMID- 21618396 TI - Meetings calender. PMID- 21618397 TI - A one-stop approach to the management of soft tissue and degenerative musculoskeletal conditions using clinic-based ultrasonography. AB - BACKGROUND AND OBJECTIVE: One-stop clinics aim at assessment, investigation and initiating treatment in a single hospital visit. They have been proven to be cost effective and to increase patient satisfaction in various specialties. The aim of this study was to describe the one-stop approach to managing soft tissue and degenerative musculoskeletal conditions using clinic-based musculoskeletal ultrasonography (MSUS). METHODS: A retrospective case record review was carried out of patients assessed and managed in the musculoskeletal clinic by a musculoskeletal and sports physician over a 10-month period. RESULTS: A total of 1,166 patients were assessed and managed in a total of 155 outpatient clinics. The age range of patients was 19 to 85 years (median age 45 years). The diagnoses included traumatic or overuse tendinopathy, degenerative arthritis, bursitis, acute/chronic sporting injuries and acute/chronic back pain. A total of 1,012 patients (87%) had conditions related to the appendicular system (shoulder girdle, upper limb, pelvic girdle and lower limb) and 154 patients were referred with spinal pain. All patients with appendicular system problems had a definite diagnosis and treatment initiated on the first visit to the clinic. In 658 (65%) patients, a diagnostic ultrasound was deemed an appropriate investigation to improve the accuracy of diagnosis and more than half of them (352 patients) needed ultrasound-guided injections as part of the management of their conditions. A portable GE Healthcare LOGIQe machine with a 10 MHz linear probe and colour Doppler flow imaging was used to perform all scans. No adverse incidents were reported. CONCLUSIONS: The use of clinic-based MSUS enables a one stop approach, reduces repeated hospital appointments and improves quality of care in an outpatient musculoskeletal clinic. PMID- 21618398 TI - The recognition and assessment of cardiovascular risk in people with rheumatoid arthritis in primary care: a questionnaire-based study of general practitioners. AB - OBJECTIVES: To investigate how well recognized the association between rheumatoid arthritis (RA) and excess cardiovascular (CV) risk is within primary care and the current assessment strategies being employed by general practitioners (GPs). METHODS: Questionnaires were sent to all 376 GPs in the Worcestershire Primary Care Trust. RESULTS: Thirty-two per cent of GPs identified RA as an independent risk factor for CV disease. Fifteen per cent and 34%, respectively, assessed their RA patients for primary and secondary prevention of their CV risks. Of those GPs who made an assessment, 18.4% adjusted the calculated risk derived from standard charts. The frequency of assessment was greater among GPs who had received a form of education about the association between CV disease and RA. However, of the GPs identifying this susceptibility, only 40% performed any form of primary prevention risk assessment. CONCLUSIONS: At present, the excess risk of CV disease conferred by RA is under-recognized and under-assessed in primary care. Currently, educational resources on this topic targeted at GPs are lacking and may in part account for our findings. However, even when GPs did identify the risk of CV disease in RA or had received education about it, this did not consistently change their clinical management. Further work to promote knowledge and management strategies for CV disease in RA is therefore needed to improve the care of patients with this condition. PMID- 21618399 TI - Physical activity coaching of patients with rheumatoid arthritis in everyday practice: a long-term follow-up. AB - OBJECTIVES: To investigate the long-term effects on perceived general health, disease activity, pain, activity limitation and cognitive behavioural factors of a one-year coaching programme performed in ordinary physical therapy practice to promote the adoption of health-enhancing physical activity in patients with early rheumatoid arthritis (RA). METHODS: A total of 228 patients with early RA, from 10 rheumatology clinics in Sweden, were randomly assigned to an intervention group (IG; n = 94) or a control group (CG; n = 134). The IG was coached by physical therapists during the first year to adopt health-enhancing levels of physical activity (30 minutes/day, moderately intensive, >= 4 days/week). No coaching was given during the subsequent year between post-intervention and follow-up. Follow-up assessment consisted of a postal questionnaire on physical activity and of visual analogue scales for ratings of general health perception and pain. The Health Assessment Questionnaire Disability Index (HAQ) and the Disease Activity Score in 28 joints (DAS 28) were collected at regular medical check-ups. RESULTS: Sixty-five (69%) participants in the IG and 92 (69%) in the CG completed the entire study period by filling in the follow-up questionnaire on physical activity two years after baseline. The intervention seemed to lack any significant influence on long-term outcome. However, different patterns of change in physical activity behaviour were observed in the two groups. CONCLUSIONS: No long-term improvement in perceived general health or other outcomes were found in the follow-up. This may partly be because the intervention lacked several important behavioural elements for physical activity maintenance. PMID- 21618400 TI - Early transcriptional regulation by C-peptide in freshly isolated rat proximal tubular cells. AB - BACKGROUND: Clinical studies have shown that proinsulin C-peptide exerts renoprotective effects in type 1 diabetes, although the underlying mechanisms are poorly understood. As C-peptide has been shown to induce several intracellular events and to localize to nuclei, we aimed to determine whether gene transcription is affected in proximal tubular kidney cells, and if so, whether the genes with altered transcription include those related to protective mechanisms. METHODS: The effect of C-peptide incubation (2 h) on gene expression was investigated in freshly isolated proximal tubular cells from streptozotocin diabetic Sprague-Dawley rats using global gene expression profiling and real-time quantitative polymerase chain reaction. Protein expression was assayed using western blotting. Different bioinformatic strategies were employed. RESULTS: Gene transcription profiling demonstrated differential transcription of 492 genes (p < 0.01) after 2 h of C-peptide exposure, with the majority of these genes repressed (83%). Real-time quantitative polymerase chain reaction validation supported a trend of several G protein-coupled receptors being activated, and certain transcription factors being repressed. Also, C-peptide repressed the transcription of genes associated with the pathways of circulatory and inflammatory diseases. CONCLUSION: This study shows that C-peptide exerts early effects on gene transcription in proximal tubular cells. The findings also bring further knowledge to the renoprotective mechanisms of C-peptide in type 1 diabetes, and support a transcriptional activity for C-peptide. It is suggested that C-peptide may play a regulatory role in the gene expression of proximal tubular cells. PMID- 21618401 TI - Structural heterogeneity and ligand gating in ferric Methanosarcina acetivorans protoglobin mutants. AB - Protoglobin from Methanosarcina acetivorans C2A (MaPgb), a strictly anaerobic methanogenic Archaea, displays peculiar structural and functional properties within members of the hemoglobin superfamily. In fact, MaPgb-specific loops and a N-terminal extension (20 amino acid residues) completely bury the heme within the protein matrix. Therefore, the access of diatomic gaseous molecules to the heme is granted by two apolar tunnels reaching the heme distal site from locations at the B/G and B/E helix interfaces. The presence of two tunnels within the protein matrix could be partly responsible for the slightly biphasic ligand binding behavior. Unusually, MaPgb oxygenation is favored with respect to carbonylation. Here, the crucial role of Tyr(B10)61 and Ile(G11)149 residues, located in the heme distal site and lining the protein matrix tunnels 1 and 2, respectively, on ligand binding to the heme-Fe-atom and on distal site structural organization is reported. In particular, tunnel 1 accessibility is modulated by a complex reorganization of the Trp(B9)60 and Phe(E11)93 side-chains, triggered by mutations of the Tyr(B10)61 and Ile(G11)149 residues, and affected by the presence and type of the distal heme-bound ligand. PMID- 21618402 TI - Oxygen binding to Arabidopsis thaliana AHb2 nonsymbiotic hemoglobin: evidence for a role in oxygen transport. AB - Nonsymbiotic hemoglobins AHb1 and AHb2 discovered in Arabidopsis thaliana are likely to carry out distinct physiological roles, in consideration of their differences in sequence, structure, expression pattern, and tissue localization. Despite a relatively fast autoxidation in the presence of O(2) , we were able to collect O(2) -binding curves for AHb2 in the presence of a reduction enzymatic system. AHb2 binds O(2) noncooperatively with a p50 of 0.021 +/- 0.003 Torr, a value consistent with a recently proposed role in O(2) transport. The analysis of the internal cavities derived from the structures sampled in molecular dynamics simulations confirms strong differences with AHb1, proposed to work as a NO deoxygenase in vivo. Overall, our results are consistent with a role for AHb2 as an oxygen carrier, as recently proposed on the basis of experiments on AHb2 overexpressing mutants of A. thaliana. PMID- 21618403 TI - The carbon monoxide-releasing molecule, CORM-3 (RU(CO)(3) CL(glycinate)), targets respiration and oxidases in Campylobacter jejuni, generating hydrogen peroxide. AB - Carbon monoxide (CO) is a classical respiratory inhibitor, but CO-releasing molecules (CO-RMs) have therapeutic value, increasing phagocytosis, and reducing sepsis-induced lethality. CORM-3, Ru(CO)(3) Cl(glycinate), a ruthenium-based carbonyl that liberates CO under physiological conditions, has previously been shown to inhibit bacterial growth and respiration, even at high concentrations of oxygen. Here, we report the effects of CORM-3 on the microaerophilic foodborne pathogen Campylobacter jejuni. Even at CO-RM (i.e., CO) concentrations that exceed dissolved oxygen levels, CORM-3 does not inhibit microaerobic growth. This insensitivity is not due to failure of CORM-3 to penetrate cells, as revealed by assay with extracellular myoglobin and by the ability of CO from externally added CORM-3 to bind intracellular membrane-associated respiratory oxidases. Even at almost 200 MU M oxygen, CORM-3 inhibits formate-dependent respiration and leads to generation of hydrogen peroxide. This work shows that CO-RMs have valuable properties as antimicrobial agents; however, growth inhibition does not always accompany inhibition of respiration, even when ambient oxygen concentrations are low. PMID- 21618404 TI - Growth variation among Bacillus thuringiensis strains can affect screening procedures for supernatant-secreted toxins against insect pests. AB - BACKGROUND: Supernatant-secreted proteins in Bacillus thuringiensis (Berliner) (Bt) with insecticidal activity provide an important source of information for discovery of new useful strains and/or entomotoxins. However, physiological variation among isolates might interfere in the detection efficiency of screening procedures on Bt collections. The aim of this study was to assess the magnitude of this variation in a sample of isolates from a tropical Bt collection, which was gauged through the assessment of their temporal patterns of growth and protein secretion in culture supernatants (SNs), as well as of the corresponding toxicity against fall armyworm (Spodoptera frugiperda, JE Smith). Feeding bioassays were performed, with larvae being treated with heated and non-heated total protein extracted from SNs collected at different culture times. Larva mortality and reduction in pupa formation were observed. RESULTS: Intra- and interisolate variations were observed in the temporal patterns of growth, quality and quantity of protein secreted, as well as in insecticidal activity of these SNs, based on larvae mortality and pupation rates. These results suggest that the insecticidal potential of certain isolates can be hidden if comparisons are done on the basis of the same number of cells in the culture and/or the same culturing time. CONCLUSIONS: Methods of screening Bt collections on the basis of feeding bioassays can be misleading with regards to identifying more promising isolates for biocontrol purposes if physiological differences are not considered. The consequences and implications of these findings for the development of experimental systems that depend on toxicity bioassays to identify alternative Bt strains and entomotoxins with practical applicability have been discussed. PMID- 21618405 TI - Epigenetic regulation in neural crest development. AB - The neural crest (NC) is a multipotent, migratory cell population that arises from the developing dorsal neural fold of vertebrate embryos. Once their fates are specified, neural crest cells (NCCs) migrate along defined routes and differentiate into a variety of tissues, including bone and cartilage of the craniofacial skeleton, peripheral neurons, glia, pigment cells, endocrine cells, and mesenchymal precursor cells (Santagati and Rijli,2003; Dupin et al.,2006; Hall,2009). Abnormal development of NCCs causes a number of human diseases, including ear abnormalities (including deafness), heart anomalies, neuroblastomas, and mandibulofacial dysostosis (Hall,2009). For more than a century, NCCs have attracted the attention of geneticists and developmental biologists for their stem cell-like properties, including self-renewal and multipotent differentiation potential. However, we have only begun to understand the underlying mechanisms responsible for their formation and behavior. Recent studies have demonstrated that epigenetic regulation plays important roles in NC development. In this review, we focused on some of the most recent findings on chromatin-mediated mechanisms for vertebrate NCC development. PMID- 21618407 TI - Heterozygous M1V variant of ELA-2 gene mutation associated with G-CSF refractory severe congenital neutropenia. AB - Severe congenital neutropenia is an autosomal recessive disorder characterized by maturation arrest at the promyelocyte/myelocyte phase in the bone marrow, absolute neutrophil count <0.5 * 10(9) /L and recurrent bacterial infections. Homozygous mutations of either HAX-1 or ELA-2 have been described. We report the case of a premature male infant with congenital neutropenia, associated with multiple infections, refractory to treatment with granulocyte colony stimulating factor who subsequently underwent matched sibling donor stem-cell transplant. He was found to be heterozygous for the M1V variant of the ELA-2 gene that we postulate to be causative for his severe neutropenia PMID- 21618406 TI - Extracellular matrix and heart development. AB - The extracellular matrix (ECM) of the developing heart contains numerous molecules that form a dynamic environment that plays an active and crucial role in the regulation of cellular events. ECM molecules found in the heart include hyaluronan, fibronectin, fibrillin, proteoglycans, and collagens. Tight regulation of the spatiotemporal expression, and the proteolytic processing of ECM components by proteases including members of the ADAMTS family, is essential for normal cardiac development. Perturbation of the expression of genes involved in matrix composition and remodeling can interfere with a myriad of events involved in the formation of the four-chambered heart and result in prenatal lethality or cardiac malformations as seen in humans with congenital heart disease. In this review, we summarize what is known about the specific importance of some of the components of the ECM in relation to the cardiovascular development. PMID- 21618408 TI - Congenital cytomegalovirus infection and Wiskott-Aldrich syndrome successfully treated with unrelated cord blood transplantation. AB - We report a successful umbilical cord blood transplantation (UCBT) in an 8-month male with Wiskott-Aldrich syndrome (WAS) and congenital cytomegalovirus (CMV) infection. The child presented at 3 months of age with symptomatic thrombocytopenia and CMV infection. Despite appropriate antiviral treatment no rise in the platelet count was observed. Genetic analysis confirmed the diagnosis of WAS. The clinical course was complicated by severe CMV retinitis with bilateral retinal hemorrhages and renal vasculitis. He underwent unrelated UCBT resulting in a rapid resolution of autoimmunity and thrombocytopenia. PMID- 21618409 TI - Risk factors for smoking among adolescent survivors of childhood cancer: a report from the Childhood Cancer Survivor Study. AB - BACKGROUND: Few studies have examined risk factors for smoking among adolescent survivors of childhood cancer. The present study reports on the rate of smoking and identifies factors associated with smoking in a sample of adolescent survivors from the Childhood Cancer Survivor Study (CCSS). PROCEDURE: Participants included 307 adolescent survivors and 97 healthy siblings (ages 14 20) who completed a self-report survey of health, quality of life, and health behaviors. RESULTS: Smoking rates did not differ significantly between survivor and sibling groups (ever smokers: 28% vs. 33%, recent smokers: 10% vs. 9%, respectively). Ever smoking was significantly associated with peer smoking, smokers in the household, binging, suicidal behavior, and no history of CRT. There were significant interactions of peer smoking with gender and CRT for ever smoking and with binging for recent smoking. Recent smoking was more likely for survivors with other household smokers (RR=2.24, CI=1.21-4.16), past suicidality (RR=1.89, CI=1.00-3.56), and no CRT (RR=2.40, CI=1.12-5.17). Among survivors with few smoking friends, ever smoking was more likely for survivors with no CRT (RR=4.47, CI=1.43-13.9), and recent smoking was more likely among survivors who binged (RR=3.37, CI=1.17-9.71). CONCLUSIONS: Despite the health risks associated with survivorship, nearly one in three adolescent survivors of childhood cancer has smoked. Exposure to other smokers, in particular, appears to increase the likelihood of smoking for some survivors. Providing smoking cessation programs targeted to family members, helping survivors choose non-smoking friends, and teaching ways to resist smoking influences from peers may be important pathways for smoking prevention with adolescent survivors. PMID- 21618410 TI - Folate pathway polymorphisms predict deficits in attention and processing speed after childhood leukemia therapy. AB - BACKGROUND: Neurocognitive impairment occurs in 20-40% of childhood acute lymphoblastic leukemia (ALL) survivors, possibly mediated by folate depletion and homocysteine elevation following methotrexate treatment. We evaluated the relationship between folate pathway polymorphisms and neurocognitive impairment after childhood ALL chemotherapy. PROCEDURE: Seventy-two childhood ALL survivors treated with chemotherapy alone underwent a neurocognitive battery consisting of: Trail Making Tests A (TMTA) and B (TMTB), Grooved Pegboard Test Dominant-Hand and Nondominant-Hand, Digit Span subtest, and Verbal Fluency Test. We performed genotyping for: 10-methylenetetrahydrofolate reductase (MTHFR 677C>T and MTHFR 1298A>C), serine hydroxymethyltransferase (SHMT 1420C>T), methionine synthase (MS 2756 A>G), methionine synthase reductase (MTRR 66A>G), and thymidylate synthase (TSER). Student's two sample t-test and analysis of covariance were used to compare test scores by genotype. RESULTS: General impairment on the neurocognitive battery was related to MTHFR 1298A>C (P = 0.03) and MS 2756A>G (P = 0.05). Specifically, survivors with MTHFR 1298AC/CC genotypes scored, on average, 13 points lower on TMTB than those with MTHFR 1298AA genotype (P = 0.001). The MS 2756AA genotype was associated with a 12.2 point lower mean TMTA score, compared to MS 2756 AG/GG genotypes (P = 0.01). The TSER 2R/3R and 3R/3R genotypes were associated with an 11.4 point lower mean score on TMTB, compared to the TSER 2R/2R genotype (P = 0.03). Survivors with >=6 folate pathway risk alleles demonstrated a 9.5 point lower mean TMTA score (P = 0.06) and 14.5 point lower TMTB score (P = 0.002) than survivors with <6 risk alleles. CONCLUSIONS: Folate pathway polymorphisms are associated with deficits in attention and processing speed after childhood ALL therapy. PMID- 21618412 TI - Very high hypertriglyceridemia induced: is plasmapheresis needed? PMID- 21618411 TI - Hedgehog pathway activity in pediatric embryonal rhabdomyosarcoma and undifferentiated sarcoma: a report from the Children's Oncology Group. AB - BACKGROUND: Aberrant activation of the hedgehog (Hh) signaling pathway is implicated widely in both pediatric and adult malignancies. Inactivation of the Hh regulator PTCH is responsible for the Gorlin cancer predisposition syndrome. The spectrum of tumors found in Gorlin Syndrome includes basal cell carcinoma, medulloblastoma, and rarely, rhabdomyosarcoma (RMS). A previous report utilizing in situ hybridization has provided initial evidence for the expression of Hh targets GLI1 and PTCH in RMS tumors. PROCEDURE: To investigate the role of Hh pathway signaling in pediatric RMS and undifferentiated sarcoma (US) tumors, the expression of Hh pathway targets GLI1 and PTCH was measured. RNA was extracted from archival human tumor specimens collected from pediatric patients enrolled on Intergroup Rhabdomyosarcoma Study III and IV, and subjected to quantitative reverse transcriptase-polymerase chain reaction. RESULTS: Expression of GLI1 with or without PTCH was detected in substantial subsets of embryonal RMS (ERMS) and US tumors but only rarely in alveolar RMS tumors. Neither PTCH mutations nor activating SMO mutations were detected in ERMS tumors with high GLI1 expression. Microarray analysis demonstrated relative overexpression of downstream Hh targets in ERMS tumors with high or intermediate GLI1 expression. Unlike a recent report, Hh pathway activity in ERMS tumors did not correlate with a unique clinical phenotype. CONCLUSIONS: Our findings support a role for Hh pathway activation in the genesis of a subset of ERMS and US tumors. Hh signaling may represent a novel therapeutic target in affected tumors. PMID- 21618413 TI - Children's psychological distress during pediatric HSCT: parent and child perspectives. AB - BACKGROUND: Hematopoietic stem cell transplantation (HSCT) can be challenging to pediatric recipients and their families. Little is known about the recipients' psychological status as they initiate treatment and in the year afterwards. The purpose of this study is to describe the psychological status of 107 pediatric HSCT recipients from their parents' perspective, and to compare reports from parents and children in a subset of 55 children. We hypothesized that there would be discrepancies between parent and child report of child distress. PROCEDURE: Multi-site, prospective study of eligible child participants and their parents who completed selected modules from the Structured Clinical Interview for DSM-IV TR, Childhood Version (KID-SCID) the month before and one year after HSCT. Diagnoses were threshold or subthreshold. RESULTS: According to parents, nearly 30% of children had anxiety disorder both before and after HSCT; approximately half of these met threshold criteria. Agreement between parents and children for anxiety disorders was poor at baseline (kappa = -0.18, 95%CI = -0.33, -0.02) and fair at 12 months (kappa = 0.31, 95%CI = -0.04, 0.66). Agreement about mood disorders was fair at baseline (10% prevalence, kappa = 0.39, 95%CI = -0.02, 0.79) and moderate at 12 months (14% prevalence, kappa = 0.41, 95%CI = 0.02, 0.80). CONCLUSIONS: Anxiety (30%) and mood (10-14%) symptoms are common in children both before and after HSCT; parent and child reports of these symptoms do not agree. Input from parents and children is recommended to identify more accurately children who may need additional intervention during and following HSCT. PMID- 21618414 TI - Ofatumumab for a rituximab-allergic child with chronic-relapsing paraneoplastic opsoclonus-myoclonus. AB - Ofatumumab is a fully human anti-CD20 monoclonal antibody in phase II-III trials for various autoimmune and lymphoreticular diseases. We used it to treat a rituximab-allergic child with severe, chronic-relapsing, opsoclonus-myoclonus syndrome (OMS), characterized by persistent cerebrospinal fluid (CSF) B-cell expansion and T-cell dysregulation. He had relapsed despite chemotherapy, plasma exchange with immunoadsorption, and resection of ganglioneuroblastoma, detected 3 years after OMS onset. The four ofatumumab infusions (1,195 mg/m(2) total dose) were well tolerated, and CSF B-cell expansion was eliminated. No further relapses have occurred in 3 years, but he remains on low-dose ACTH with neuropsychiatric residuals of OMS. PMID- 21618415 TI - A novel mutation Gly603Arg of TMPRSS6 in a Korean female with iron-refractory iron deficiency anemia. AB - Iron-refractory iron deficiency anemia (IRIDA) is a rare hereditary form of IDA with autosomal recessive inheritance. IRIDA is characterized by hypochromic microcytic anemia unresponsive to oral iron treatment, low transferrin saturation, and a high level of iron-regulated hormone hepcidin. The genetic background of IRIDA is mutations in the TMPRSS6 gene encoding matriptase-2 (TMPRSS6) that prevent inactivation of hemojuvelin, an activator of hepcidin transcription. We herein report a Korean female with IRIDA who was compound heterozygous for two mutations in TMPRSS6: a novel missense mutation c.1807G>C (p.Gly603Arg) in the serine protease domain and a known splicing mutation c.863+1G>T (IVS6+1G>T). PMID- 21618416 TI - There's more to surviving transplant than remission. PMID- 21618418 TI - In vitro fertilization and risk of childhood leukemia in Greece and Sweden. AB - BACKGROUND: Cancer risk in children born after in vitro fertilization (IVF) remains largely unknown. We aimed to investigate risk of leukemia and lymphoma following IVF using two nationwide datasets. METHODS: The hospital-based case control study in Greece derived from the National Registry for Childhood Hematological Malignancies (1996-2008, 814 leukemia and 277 lymphoma incident cases with their 1:1 matched controls). The Swedish case-control study was nested in the Swedish Medical Birth Register (MBR) (1995-2007, 520 leukemia and 71 lymphoma cases with their 5,200 and 710 matched controls) with ascertainment of incident cancer cases in the National Cancer Register. Study-specific and combined odds ratios (OR) were estimated using conditional logistic regression, with adjustment for possible risk factors. RESULTS: Nationwide studies pointed to similar size excess risk of leukemia following IVF, but to a null association between IVF and lymphoma. The proportion of leukemia cases conceived through IVF was 3% in Greece and 2.7% in Sweden; prevalence of IVF in matched controls was 1.8% and 1.6%, respectively. In combined multivariable analyses, the increased risk of leukemia was confined to age below 3.8 years (OR = 2.21; 95% confidence interval, CI: 1.27-3.85) and to acute lymphoblastic leukemia (ALL) (OR = 1.77; 95% CI: 1.06-2.95) with no sufficient evidence of excess risk for other leukemias (OR = 1.34; 95% CI: 0.38-4.69). Following IVF, OR for ALL was 2.58 (95% CI: 1.37 4.84) before age 3.8 and 4.29 (95% CI: 1.49-12.37) before age 2 years. CONCLUSIONS: IVF seems to be associated with increased risk of early onset ALL in the offspring. PMID- 21618417 TI - Germline genetic variation and treatment response on CCG-1891. AB - BACKGROUND: Recent studies suggest that polymorphisms in genes encoding enzymes involved in drug detoxification and metabolism may influence disease outcome in pediatric acute lymphoblastic leukemia (ALL). We sought to extend current knowledge by using standard and novel statistical methodology to examine polymorphic variants of genes and relapse risk, toxicity, and drug dose delivery in standard risk ALL. PROCEDURE: We genotyped and abstracted chemotherapy drug dose data from treatment roadmaps on 557 patients on the Children's Cancer Group ALL study, CCG-1891. Fourteen common polymorphisms in genes involved in folate metabolism and/or phase I and II drug detoxification were evaluated individually and clique-finding methodology was employed for detection of significant gene gene interactions. RESULTS: After controlling for known risk factors, polymorphisms in four genes: GSTP1*B (HR = 1.94, P = 0.047), MTHFR (HR = 1.61, P = 0.034), MTRR (HR = 1.95, P = 0.01), and TS (3R/4R, HR = 3.69, P = 0.007) were found to significantly increase relapse risk. One gene-gene pair, MTRR A/G and GSTM1 null genotype, significantly increased the risk of relapse after correction for multiple comparisons (P = 0.012). Multiple polymorphisms were associated with various toxicities and there was no significant difference in dose of chemotherapy delivered by genotypes. CONCLUSIONS: These data suggest that various polymorphisms play a role in relapse risk and toxicity during childhood ALL therapy and that genotype does not play a role in adjustment of drug dose administered. Additionally, gene-gene interactions may increase the risk of relapse in childhood ALL and the clique method may have utility in further exploring these interactions. childhood ALL therapy. PMID- 21618420 TI - Cost-effectiveness of treatment of childhood acute lymphoblastic leukemia with chemotherapy only: the influence of new medication and diagnostic technology. AB - BACKGROUND: Survival for childhood acute lymphoblastic leukemia (ALL) has reached 80-90%. Future improvement in treatment success will involve new technologies and medication, adding to the pressure on limited financial resources. Therefore a retrospective cost-effectiveness analysis of ALL treatment with chemotherapy only according to the two most recent Dutch Childhood Oncology Group treatment protocols was performed. The most recent protocol ALL10 included more expensive medication (pegasparaginase) and implemented a new diagnostic technique (minimal residual disease levels) compared to the previous ALL9 protocol. PROCEDURE: Fifty children from a single center cohort were included. All direct medical costs made during treatment, including those in satellite hospitals, were determined. Costs per life year saved (LYS) were calculated. The cost-effectiveness ratio of the most recent treatment protocol was determined. LYS were calculated based on national 5-year event-free survival. RESULTS: Mean total costs were between $115,858 (ALL9) and $163,350 (ALL10) per patient. Hospital admissions (57%) and medication (11-17%) were important drivers of overall costs, and were higher in the most recent protocol ALL10. Costs per LYS were $1,962 (ALL9) and $2,655 (ALL10) and the cost-effectiveness ratio was $8,215. CONCLUSION: Treatment of childhood ALL with chemotherapy only is well within accepted ranges of cost effectiveness. The use of new technology and more expensive medication in the most recent protocol ALL10 lead to higher costs but more LYS. In future (ALL) treatment protocols, costs in relation to effects should be taken into account in order to establish more cost-effective disease management without jeopardizing survival and quality of life. PMID- 21618421 TI - Favorable outcome with conservative treatment for children with low grade brainstem tumors. AB - BACKGROUND: Pediatric brainstem tumors (BST) comprise a heterogeneous group of entities. Data regarding treatment options and outcome of BST, specifically brainstem low grade tumors (BSLGT) are limited. In order to better define risk groups and evaluate treatment options for pediatric BST, we performed a comprehensive analysis of all BST patients treated in our hospital during the MRI era. PROCEDURES: We retrospectively analyzed clinical, imaging, and pathology data at presentation, treatment, and outcome of all BST patients followed at the Hospital for Sick Children in Toronto over the last 25 years. RESULTS: Of 1,801 children with brain tumors, 223 (12%) had a brainstem primary location. Tumors without pontine involvement were BSLGT in 98.3%, whereas 75% of tumors involving the pons were high grade (P = 0.0001). Patients with BSLGT had 5-year progression free survival (PFS) and overall survival (OS) of 57 +/- 3% and 89 +/- 5%, respectively. Upfront observation of tumor residual conferred no survival disadvantage with 5-year PFS and OS of 57 +/- 5% and 93 +/- 3%, respectively. In the group of patients requiring further treatment, 5-year PFS and OS were comparable between chemotherapy and radiotherapy with 53 +/- 12% and 93 +/- 4% and 66 +/- 11% and 83 +/- 6%, respectively (P = 0.26 and 0.3, respectively). CONCLUSION: BST without pontine involvement are almost invariably BSLGT. Children with BSLGT have an excellent outcome even with careful initial observation. No clear benefit was observed for radiotherapy over chemotherapy when adjuvant treatment was needed. A conservative approach may be warranted for children with non-pontine brainstem lesions. PMID- 21618422 TI - Superior outcome of pediatric acute myeloid leukemia patients with orbital and CNS myeloid sarcoma: a report from the Children's Oncology Group. AB - BACKGROUND: Extramedullary leukemia (EML) is common in pediatric acute myeloid leukemia (AML) and occurs as leukemia cells within the cerebrospinal fluid (CSF) or as a solid tumor (myeloid sarcoma-MS). The effect of MS on survival is unknown. METHODS: Patients on CCG protocols 2861, 2891, 2941, and 2961 being treated for AML with intensive-timing chemotherapy were classified for the presence of EML (CSF leukemia, CNS-MS, orbital-MS, or non-CNS MS). CSF leukemia was classified as CNS3 (>5 WBC in the CSF with blasts) and non-CSF leukemia as CNS1/2 (<5 WBC in the CSF with or without blasts). Characteristics and outcomes of these patients were compared. RESULTS: Of the 1,459 total patients, 1,206 (82%) had no EML, 154 (11%) had CSF leukemia, 19 (1%) had CNS-MS, 23 (2%) had orbital-MS, and 57 (4%) had non-CNS MS. The CR rate was significantly higher in patients with orbital-MS and CNS-MS than in those with non-MS and non-CNS MS (96% and 95% vs. 78% and 78%, P = 0.034). Patients with orbital-MS and CNS-MS had significantly higher overall survival than patients with non-CNS MS (92% and 73% vs. 38%, P < 0.001), CNS3 patients (92% and 73% vs. 51, P < 0.001), and CNS1/2 patients (92% and 73% vs. 50%, P < 0.001). Patients with orbital-MS had a significantly lower relapse rate. CONCLUSION: Patients with MS involving orbital and CNS sites had a significantly better survival than patients with non-CNS MS, with CSF leukemia, or with no EML. PMID- 21618423 TI - Misunderstandings of the genetics and neurobiology of ADHD: moving beyond anachronisms. PMID- 21618424 TI - A microflow cytometer exploited for the immunological differentiation of leukocytes. AB - In this article, we demonstrate the potential of a microfluidic chip for the differentiation of immunologically stained blood cells. To this end, white blood cells stained with antibodies typically applied for the determination of the immune status were measured in the micro-device. Relative concentrations of lymphocytes and subpopulations of lymphocytes are compared to those obtained with a conventional flow cytometer. The stability of the hydrodynamic focusing and the optical setup was determined by measuring the variation of the signal pulse height of fluorescence calibration beads, being about 2% for the micro-device. This value and the overall performance of the micro-device are similar to conventional flow cytometers. It follows from our results that such microfluidic structures are well suited as modules in a compact, portable read-out instrument. The production process of the microflow cytometers, which we exploited for immunological cell differentiation, is compatible with mass production technology like injection molding and, hence, low cost disposable chips could be available in the future. PMID- 21618425 TI - Relationship between lunasin's sequence and its inhibitory activity of histones H3 and H4 acetylation. AB - SCOPE: Dysfunction of histone acetyltransferases (HATs) or histone deacetylases (HDACs) involved in histones acetylation has been associated with cancer. Inhibitors of these enzymes are becoming potential targets for new therapies. METHODS AND RESULTS: This study reports by Western-Blot analysis, that peptide lunasin is mainly an in vitro inhibitor of histone H4 acetylation by P300/cAMP response element-binding protein (CBP)-associated factor (PCAF), with IC50 values dependent on the lysine position sensitive to be acetylated (0.83 MUM (H4-Lys 8), 1.27 MUM (H4-Lys 12) and 0.40 MUM (H4-Lys 5, 8, 12, 16)). Lunasin is also capable of inhibiting H3 acetylation (IC50 of 5.91 MUM (H3-Lys 9) and 7.81 MUM (H3-Lys 9, 14)). Studies on structure-activity relationship establish that lunasin's sequence are essential for inhibiting H4 acetylation whereas poly-D sequence is the main active sequence responsible for H3 acetylation inhibition. Lunasin also inhibits H3 and H4 acetylation and cell proliferation (IC50 of 181 MUM) in breast cancer MDA-MB-231 cells. Moreover, this peptide decreases expression of cyclins and cyclin dependent kinases-4 and -6, implicated in cell cycle pathways. CONCLUSION: Results from this study demonstrates lunasin's role as modulator of histone acetylation and protein expression that might contribute on its chemopreventive properties against breast cancer. PMID- 21618426 TI - Urinary N-methylpyridinium and trigonelline as candidate dietary biomarkers of coffee consumption. AB - SCOPE: In order to validate the in vivo function of putatively healthy molecules in foods, human intervention studies are required. As the subject's compliance concerning intake or abstinence of a given food is considered mandatory to be monitored by biomarkers, the objective was to identify analytical markers for coffee consumption. METHODS AND RESULTS: Urine samples collected from coffee drinkers were compared with those of non-coffee drinkers using hydrophilic liquid interaction chromatography (HILIC)/time-of-flight mass spectrometry-based metabolite profiling. Two urinary molecules, found to be contributing most to the dissimilarities between both groups, were identified as N-methylpyridinium (NMP) and trigonelline and their suitability as coffee-specific biomarkers was validated by means of a coffee intervention study. After the volunteers (five females and four males) consumed a single dose of coffee, morning urine was collected for 10 days while staying abstinent from any coffee. HILIC-MS/MS-stable isotope dilution analysis (SIDA) revealed elevated urinary concentrations of trigonelline and NMP for up to 48 (p=0.001) and 72 h (p=0.002), respectively, after coffee consumption when compared with non-coffee drinkers. CONCLUSION: Analysis of urinary NMP allows to check for coffee consumption within a period of 3 days and is proposed as a dietary biomarker which might be used as an analytical probe to control compliance in human intervention studies on coffee. PMID- 21618427 TI - Long-term adaptation of global transcription and metabolism in the liver of high fat diet-fed C57BL/6J mice. AB - SCOPE: This study investigated the global transcriptional and metabolic changes occurring at multiple time points over 24 wk in response to a high-fat diet (HFD). METHODS AND RESULTS: C57BL/6J mice were fed a HFD or normal diet (ND) over 24 wk. HFD-fed mice developed early clinical indicators of obesity-related co morbidities including fatty liver, insulin resistance, hyperglycemia and hypercholesterolemia. Time-course microarray analysis at eight time points over 24 wk identified 332 HFD responsive genes as potential targets to counteract diet induced obesity (DIO) and related co-morbidities. Glucose regulating enzyme activity and gene expression were altered early in the HFD-fed mice. Fatty acid (FA) and triglyceride (TG) accumulation in combination with inflammatory changes appear to be likely candidates contributing to hepatic insulin resistance. Cidea seemed to be one of representative genes related to these changes. CONCLUSION: Global transcriptional and metabolic profiling across multiple time points in liver revealed potential targets for nutritional interventions to reverse DIO. In future, new approaches targeting HFD responsive genes and hepatic metabolism could help ameliorate the deleterious effects of an HFD and DIO-related complication. PMID- 21618428 TI - Fabrication of advanced particles and particle-based materials assisted by droplet-based microfluidics. AB - Recent advances in the fabrication of complex particles and particle-based materials assisted by droplet-based microfluidics are reviewed. Monodisperse particles with expected internal structures, morphologies, and sizes in the range of nanometers to hundreds of micrometers have received a good deal of attention in recent years. Due to the capability of generating monodisperse emulsions and of executing precise control and operations on the suspended droplets inside the microchannels, droplet-based microfluidic devices have become powerful tools for fabricating complex particles with desired properties. Emulsions and multiple emulsions generated in the microfluidic devices can be composed of a variety of materials including aqueous solutions, gels, polymers and solutions containing functional nanoparticles. They are ideal microreactors or fine templates for synthesizing advanced particles, such as polymer particles, microcapsules, nanocrystals, and photonic crystal clusters or beads by further chemical or physical operations. These particles are promising materials that may be applicable for many fields, such as photonic materials, drug delivery systems, and bio-analysis. From simple to complex, from spherical to nonspherical, from polymerization and reaction crystallization to self-assembly, this review aims to help readers be aware of the many aspects of this field. PMID- 21618429 TI - Automated on-fiber derivatization with headspace SPME-GC-MS-MS for the determination of primary amines in sewage sludge using pressurized hot water extraction. AB - An automated, environmentally friendly, simple, selective, and sensitive method was developed for the determination of ten primary aliphatic amines in sewage sludge at MUg/kg dry weight (d.w.). The procedure involves a pressurized hot water extraction (PHWE) of the analytes from the solid matrix, followed by a fully automated on-fiber derivatization with 2,3,4,5-pentafluorobenzaldehyde (PFBAY) and headspace solid-phase microextraction (HS-SPME) and subsequent gas chromatography ion-trap tandem mass spectrometry (GC-IT-MS-MS) analysis. The limits of detection (LODs) of the method were between 0.5 and 45 MUg/kg (d.w.) for all compounds except for ethyl-, isopropyl-, and amylamine, whose LODs were 70, 109, and 116 MUg/kg (d.w.), respectively. The limits of quantification (LOQs) were between 10 and 350 MUg/kg (d.w.). Repeatability and intermediate precision, expressed as RSD(%) (n=3), were lower than 18 and 21%, respectively. The method developed enabled to determine primary aliphatic amines in sludge from various urban and industrial sewage treatment plants as well as from a potable treatment plant. Most of the primary aliphatic amines were found in the sewage sludge samples analyzed corresponding to the maximum concentrations to the samples from the urban plant: for instance, isobutylamine and methylamine were found at 7728 and 12 536 MUg/kg (d.w.), respectively. Amylamine was detected only in few samples but always at concentrations lower than its LOQ. PMID- 21618430 TI - Development of a new open-tubular capillary electrochromatography method for in vitro monitoring of toxic aromatic amines distribution in rat blood. AB - Exposure to aromatic amines from different industrial and agricultural activities entails a substantial risk of deleterious somatic effects, genetic damage and cancer development. Thus, a new and simple method for separation and analysis of aromatic amines has been developed by open-tubular capillary electrochromatography with a novel amphipathic block copolymer (poly(tert-butyl acrylate)(127)-block-poly(glycidyl methacrylate)(86)) coating based on its self assembled property. Key factors affecting the separation efficiency of the test analytes, such as pH, buffer concentration and selective solvent, were studied in detail. Meanwhile, method validation was well evaluated by linearity (>=0.998), detection limit and recovery. Application of this developed protocol on in vitro monitoring of the target aromatic amines distribution in rat blood demonstrated its potential usage for separation and determination of aromatic amines in biological samples. Additionally, for assimilating more polymeric materials into analysis of aromatic amines, the effect of morphology changes of the amphipathic block copolymer coating on open-tubular capillary electrochromatography separation was also studied, and the result revealed that the block copolymer coating could play the same role as surfactant. PMID- 21618431 TI - Noise normalisation in capillary electrophoresis using a diode array detector. AB - When using capillary electrophoresis with a diode array detector, the wavelength at maximum absorbance is often chosen to quantify a given analyte. However, the background noise for every wavelength should be taken into account as it is by maximising the signal to noise ratio that the lowest limit of detection will be obtained. Here, we proposed an algorithm allowing to correct an electropherogram from its background absorption and to estimate the background noise. Applying it to all the electropherograms obtained in each wavelength channel allows obtaining the background noise as a function of the wavelength, which can be used to calculate the signal to noise ratio. This not only allows selecting the best wavelength to maximise the limit of detection of a given analyte, but also to generate a noise normalised base peak electropherogram (nn-BPE). It is shown that the noise normalised base peak electropherograms substantially improve the peaks visualisation. The algorithm is part of a graphic user interface that runs under MatLab environment; it does not require any programming knowledge and is freely available. PMID- 21618432 TI - A 18F-labeled fluorobutyl-substituted spirocyclic piperidine derivative as a selective radioligand for PET Imaging of sigma1 receptors. AB - In this study, we synthesized and evaluated a new spirocyclic piperidine derivative 3, containing a 4-fluorobutyl side chain, as a PET radioligand for neuroimaging of sigma1 receptors. In vitro, compound 3 displayed high affinity for sigma1 receptors (K(i) =1.2 nM) as well as high selectivity. [18F]3 radiosynthesis was performed from the corresponding tosylate precursor, with high radiochemical yield (45-51 %), purity (>98 %), and specific activity (>201 GBq MUmol-1). Metabolic stability of [18F]3 in the brain of CD-1 mice was verified, and no penetration of peripheral radiometabolites into the cerebral tissue was observed. Results of ex vivo autoradiography revealed that the distribution of [18F]3 in the brain corresponded to regions with high sigma1 receptor density. The highest region-specific total-to-nonspecific ratio was determined in the facial nucleus (4.00). Biodistribution studies indicated rapid and high levels in brain uptake of [18F]3 (2.2 % ID per gram at 5 min p.i.). Pre-administration of haloperidol significantly inhibited [18F]3 uptake into the brain and sigma1 receptor-expressing organs, further confirming in vivo target specificity. PMID- 21618433 TI - Antagonism of the Stat3-Stat3 protein dimer with salicylic acid based small molecules. AB - More than 50 new inhibitors of the oncogenic Stat3 protein were identified through a structure-activity relationship (SAR) study based on the previously identified inhibitor S3I-201 (IC50 =86 MUM, K(i) >300 MUM). A key structural feature of these inhibitors is a salicylic acid moiety, which, by acting as a phosphotyrosine mimetic, is believed to facilitate binding to the Stat3 SH2 domain. Several of the analogues exhibit higher potency than the lead compound in inhibiting Stat3 DNA binding activity, with an in vitro IC50 range of 18.7-51.9 MUM, and disruption of Stat3-pTyr peptide interactions with K(i) values in the 15.5-41 MUM range. One agent in particular exhibited potent inhibition of Stat3 phosphorylation in both breast and multiple myeloma tumor cells, suppressed the expression of Stat3 target genes, and induced antitumor effects in tumor cells harboring activated Stat3 protein. PMID- 21618434 TI - Synthesis of soybean oil-based thiol oligomers. AB - Industrial grade soybean oil (SBO) and thiols were reacted to generate thiol functionalized oligomers via a thermal, free radical initiated thiol-ene reaction between the SBO double bond moieties and the thiol functional groups. The effect of the reaction conditions, including thiol concentration, catalyst loading level, reaction time, and atmosphere, on the molecular weight and the conversion to the resultant soy-thiols were examined in a combinatorial high-throughput fashion using parallel synthesis, combinatorial FTIR, and rapid gel permeation chromatography (GPC). High thiol functionality and concentration, high thermal free radical catalyst concentration, long reaction time, and the use of a nitrogen reaction atmosphere were found to favor fast consumption of the SBO, and produced high molecular weight products. The thiol conversion during the reaction was inversely affected by a high thiol concentration, but was favored by a long reaction time and an air reaction atmosphere. These experimental observations were explained by the initial low affinity of the SBO and thiol, and the improved affinity between the generated soy-thiol oligomers and unreacted SBO during the reaction. The synthesized soy-thiol oligomers can be used for renewable thiol-ene UV curable materials and high molecular solids and thiourethane thermal cure materials. PMID- 21618435 TI - Development of the gubernaculum during testicular descent in the rat. AB - Gubernacular elongation during inguinoscrotal testicular descent and cremaster muscle development remains poorly described in mammals. The role of the genitofemoral nerve (GFN) remains elusive. We performed detailed histological analysis of testicular descent in normal rats to provide a comprehensive anatomical description for molecular studies. Fetuses and neonatal male offspring (5-10 per group) from time-mated Sprague-Dawley dams (embryonic days 15, 16, and 19; postnatal days 0, 2, and 8) were prepared for histology. Immunohistochemistry was performed for nerves (Class III tubulin, Tuj1) and muscle (desmin). At embryonic days 15 and 16, the gubernaculum and breast bud are adjacent and both supplied by the GFN. By embryonic day 19, the breast bud has regressed and the gubernacular swelling reaction is completed. Postnatally, the gubernacular core regresses, except for a cranial proliferative zone. The cremaster is continuous with internal oblique and transversus abdominis. By postnatal day 2 (P2), the gubernaculum has everted, locating the proliferative zone caudally and the residual mesenchymal core externally. Eversion creates the processus vaginalis, with the everted gubernaculum loose in subcutaneous tissue but still remote from the scrotum. By P8, the gubernaculum has nearly reached the scrotum with fibrous connections attaching the gubernaculum to the scrotal skin. A direct link between GFN, gubernaculum, and breast bud suggests that the latter may be involved in gubernacular development. Second, the cremaster muscle is continuous with abdominal wall muscles, but most of its growth occurs in the distal gubernacular tip. Finally, gubernacular eversion at birth brings the cranial proliferative zone to the external distal tip, enabling gubernacular elongation similar to a limb bud. PMID- 21618436 TI - Ontogenetic change in temporal bone pneumatization in humans. AB - Pneumatization of the temporal bone is often included in descriptions of fossils and as a phylogenetic marker, but a number of questions about the evolution, growth, and development of the trait remain. Many studies have analyzed temporal bone pneumatization from a clinical perspective, but a systematic quantification of normal development of pneumatized spaces has not been conducted. In this study, ontogenetic change in the size and organization of temporal bone pneumatization is analyzed in a cross-sectional sample of humans. High resolution computed tomography scans of the temporal bone were acquired from a cross sectional sample of humans (N = 28). Bone volume fractions, anisotropy, trabecular number, trabecular thickness, surface area, and volume were analyzed to provide information about the organization and size of pneumatized spaces across ontogeny. The results indicate that there are general and region-specific patterns of ontogenetic changes in the organization of pneumatized spaces. These changes reflect the transition from nonpneumatized bone to pneumatized bone. It also demonstrates that those regions that are pneumatized early in ontogeny (such as the mastoid antrum) continue to remodel after the initial period of pneumatization. The dynamic nature of temporal bone pneumatization over ontogeny suggests that this character requires careful consideration when used as a character for phylogenetic analyses. These results demonstrate the importance of comparing individuals from similar developmental stages, especially when completing quantitative analyses of the extent of pneumatization or organization of the spaces. PMID- 21618437 TI - Characterization of the spatial arrangement of secondary osteons in the diaphysis of equine and canine long bones. AB - The blood supply of bone cells in compact bone is provided primarily by blood vessels located within Haversian canals forming the centre of osteons. Mid diaphysial cross-sections of radii and third metacarpal bones from two horses and radii from two mature dogs were studied using reflective light microscopy to quantify the spatial ordering of canals and compared to a computational model. The distributions of canals were analyzed using: 1) the autocorrelation function (ACF), which describes the probability of finding two canals separated by a given distance and 2) the shortest distance distribution (SDD), which describes the probability that a site within bone is located at a given distance from the nearest canal. The order in the investigated horse radii, as characterized by the oscillations of the ACF, was found to be independent of the anatomical location although, in the metacarpal bone the order was higher in the lateral than in the cranial location. Among the dogs, marked differences were only found in the ACF. An analysis of the SDD demonstrates that ordering of canals minimizes the distance of osteocytes from a blood vessel. This suggests that the efficiency of the blood supply can be adapted through differences in the order of the Haversian canals. In our model, the ordering of canals is achieved via an exclusion zone around each canal, imposed upon newly formed osteons. Simulations demonstrate that differences in the observed order can be explained by either a larger size or a larger variability of this exclusion zone. PMID- 21618438 TI - Convergent evolution driven by similar feeding mechanics in balaenopterid whales and pelicans. AB - The feeding apparatuses of rorqual whales and pelicans exhibit a number of similarities, including long, kinetic jaws that increase gape size, and extensible tissue comprising the floor of the mouth. These specializations enable the engulfment of large volumes of prey-laden water in both taxa. However, the mechanics of engulfment feeding in rorquals and pelicans have never been quantitatively compared. Here, we use "BendCT," a novel analytical program, to investigate the mechanical design of rorqual and pelican mandibles, to understand whether these bones show comparable designs for resisting similar hydrodynamical loads. We also compare the mechanical properties of the extensible tissue used during engulfment in rorquals and pelicans. We demonstrate that the evolutionary convergence in the feeding apparatus of rorquals and pelicans is more pronounced than has been recognized previously; both taxa exhibit mandibular flexural rigidity distributions suited for resisting dorsoventral bending stresses encountered while feeding, and possess similarly extensible tissue on the floor of their mouths. PMID- 21618439 TI - Chick amniogenesis is mediated by an actin cable. AB - This study examined the process of chick amniogenesis to determine whether the actin cable mechanism operating during amnion wound repair is a recapitulation of developmental events. Staining of the developing amnion with fluorescein isothiocyanate-labeled phalloidin indicated the presence of an actin cable in the amniotic head fold, which persisted through to the closure of the amnion. Transmission electron microscopy of the developing amnion revealed linearly arranged actin microfilaments in the elongated cells at the leading edge of the amnion, adjacent to either side of a nodule of numerous cells at the point of midline fusion. A mesh of cytoplasmic actin filaments was seen dispersed throughout the accumulated cells of the nodule. Lamellapodia were absent suggesting that cell crawling is not involved in amniogenesis. Addition of an enzyme inhibitor of Rho, cell-permeable C3 transferase, to the surface of the developing amnion prior to closure appeared to inhibit amniogenesis at the early embryonic stages. PMID- 21618440 TI - Effect of monochromatic light on melatonin secretion and arylalkylamine N acetyltransferase mRNA expression in the retina and pineal gland of broilers. AB - The goal of this study is to investigate the effects of various monochromatic lights on plasma melatonin (MT) levels and the expression of arylalkylamine N acetyltransferase (AANAT) mRNA in the pineal gland and retina. A total of 160 newly hatched (posthatching day 1, P1) broilers, including intact, sham-operated, and pinealectomized groups were exposed to blue light (BL), green light (GL), red light (RL), and white light (WL) by light emitting diode (LED) system for short term (24 hr) or long term (2 weeks), separately. For intact and sham-operated birds, the plasma MT level exhibited marked circadian rhythms at P7 and P14 regardless of short-term and long-term exposure to four monochromatic lights. However, WL and BL showed a faint suppression of MT secretion in contrast to GL and RL at either light or dark time points, with the following rank order: GL < RL < WL < BL. Larger circadian amplitude of MT levels was observed in GL group versus BL group (at P14: 87.70 pg/mL vs. 19.85 pg/mL, respectively). Pinealectomy disturbed the MT rhythm under different light colors, especially in RL. Additionally, consistent with the alteration of plasma MT levels, we observed increased AANAT mRNA expression and immunoreactive cell numbers of proliferating cell nuclear antigen (PCNA) and c-Fos in the pineal gland or retina in GL than that of BL, whereas 5-HT immunoreactive cell number was significantly decreased in GL. These data suggested that GL enhanced chick pinealocytes and retinal cells to express AANAT mRNA and to secrete MT, which may be depended on promoting c-Fos expression and cell proliferation. PMID- 21618441 TI - Comparing 3D virtual methods for hemimandibular body reconstruction. AB - Reconstruction of fractured, distorted, or missing parts in human skeleton presents an equal challenge in the fields of paleoanthropology, bioarcheology, forensics, and medicine. This is particularly important within the disciplines such as orthodontics and surgery, when dealing with mandibular defects due to tumors, developmental abnormalities, or trauma. In such cases, proper restorations of both form (for esthetic purposes) and function (restoration of articulation, occlusion, and mastication) are required. Several digital approaches based on three-dimensional (3D) digital modeling, computer-aided design (CAD)/computer-aided manufacturing techniques, and more recently geometric morphometric methods have been used to solve this problem. Nevertheless, comparisons among their outcomes are rarely provided. In this contribution, three methods for hemimandibular body reconstruction have been tested. Two bone defects were virtually simulated in a 3D digital model of a human hemimandible. Accordingly, 3D digital scaffolds were obtained using the mirror copy of the unaffected hemimandible (Method 1), the thin plate spline (TPS) interpolation (Method 2), and the combination between TPS and CAD techniques (Method 3). The mirror copy of the unaffected hemimandible does not provide a suitable solution for bone restoration. The combination between TPS interpolation and CAD techniques (Method 3) produces an almost perfect-fitting 3D digital model that can be used for biocompatible custom-made scaffolds generated by rapid prototyping technologies. PMID- 21618442 TI - Masticatory hypermuscularity is not related to reduced cranial volume in myostatin-knockout mice. AB - It has been suggested recently that masticatory muscle size reduction in humans resulted in greater encephalization through decreased compressive forces on the cranial vault. Following this logic, if masticatory muscle size were increased, then a reduction in brain growth should also occur. This study was designed to test this hypothesis using a myostatin (GDF-8) knockout mouse model. Myostatin is a negative regulator of skeletal muscle growth, and individuals lacking this gene show significant hypermuscularity. Sixty-two [32 wild-type (WT) and 30 GDF-8 -/- knockout], 1, 28, 56, and 180-day-old CD-1 mice were used. Body and masseter muscle weights were collected following dissection and standardized lateral and dorsoventral cephalographs were obtained. Cephalometric landmarks were identified on the radiographs and cranial volume was calculated. Mean differences were assessed using a two-way ANOVA. KO mice had significantly greater body and masseter weights beginning at 28 days compared with WT controls. No significant differences in cranial volumes were noted between KO and WT. Muscle weight was not significantly correlated with cranial volume in 1, 28, or 180-day-old mice. Muscle weights exhibited a positive correlation with cranial volume at 56 days. Results demonstrate that masticatory hypermuscularity is not associated with reduced cranial volume. In contrast, there is abundant data demonstrating the opposite, brain growth determines cranial vault growth and masticatory apparatus only affects ectocranial morphology. The results presented here do not support the hypothesis that a reduction in masticatory musculature relaxed compressive forces on the cranial vault allowing for greater encephalization. PMID- 21618443 TI - The mechanical significance of the temporal fasciae in Macaca fascicularis: an investigation using finite element analysis. AB - Computational finite element analyses (FEAs) of the skull predict structural deformations under user specified loads and constraints, with results normally presented as stress and strain distributions over the skull's surface. The applied loads are generally a representation of the major adductor musculature, with the skull constrained at bite positions and at the articulating joints. However, virtually all analyses ignore potentially important anatomical structures, such as the fasciae that cover the temporalis muscle and attach onto the zygomatic arch. In vivo experimental studies have shown that removal of the temporal fasciae attachment onto the zygomatic arch in Cebus monkeys results in significant bone adaptation and remodeling in this region, suggesting the fasciae play an important role in stabilising the arch during biting. Here we investigate this potential stabilising role by carrying out FEAs of a macaque skull with and without temporal fasciae included. We explore the extent to which the zygomatic arch might be stabilized during biting by a synchronized tensioning of the temporal fasciae, acting to oppose masseteric contraction forces. According to our models, during temporalis muscle bulging the forces generated within the tensioned temporal fasciae are large enough to oppose the pull of the masseter. Further, a near bending-free state of equilibrium within the arch can be reached, even under forceful biting. We show that it is possible to eliminate the high strain gradients in and around the zygomatic arch that are present in past computational studies, with strains being more uniform in magnitude than previously thought. PMID- 21618444 TI - Outcomes from building system courseware for teaching and testing in a discipline based human structure curriculum. AB - This study investigated the educational benefits of system-based lecture notes and interactive learning objects in a peripheral nervous system component of a traditional first-year medical school human anatomy course. The impetus for the investigation was anecdotal evidence suggesting enhanced learner satisfaction with the learning resources. Retrospective review of existing data from 2006 to 2009 was undertaken to quantify (1) the effects of Web-based system courseware on examination item performance, and (2) differences among learner opinions regarding the benefit level of the five different types of interactive learning objects as evaluated by instructional design questionnaires. Interactive patient based case studies (IPCS) and review games (Games), simulated interactive patients (SIP), flashcards, and unit quizzes (Quizzes) developed in-house have been peer-reviewed and published in MedEdPORTAL. Statistics included one-way analysis of variance, Tukey's post hoc test, and power meta-analysis (d). Examination item analysis scores remained significantly higher (P <= 0.05; d = 0.3938) for learners receiving the instructional treatment incorporating system based lecture notes and interactive learning objects than for those not receiving this treatment. Using questionnaires with a five-point Likert scale, students reported favorably on the benefit level of all learning objects. They rated the SIP and IPCS significantly higher (P <=0.05) and games significantly lower (P <= 0.05) than in previous years, indicating a change in learner preferences. This study reaffirms that online system-based instructional techniques improve student examination performance and overall student satisfaction. Learners indicated stronger preferences for SIP and IPCS over exercises encouraging passive memorization of anatomical content. PMID- 21618445 TI - Promoting metacognition in first year anatomy laboratories using plasticine modeling and drawing activities: a pilot study of the "blank page" technique. AB - Many first year students of anatomy and physiology courses demonstrate an inability to self-regulate their learning. To help students increase their awareness of their own learning in a first year undergraduate anatomy course, we piloted an exercise that incorporated the processes of (1) active learning: drawing and plasticine modeling and (2) metacognition: planning, monitoring, reaction, and reflection. The activity was termed "blank page" because all learning cues were removed and students had to create models and diagrams from reflection and recall. Two hundred and eighty-two students responded to a questionnaire reporting qualitative feedback on the exercise. Based on student responses, the "blank page" activity was a positive learning experience and confirmed a need to teach metacognitive skills. From this pilot study, we established that drawing or plasticine modeling is an excellent vehicle for demonstration of the metacognitive processes that enable self-regulation: a known predictor of academic success. PMID- 21618446 TI - Surgeons as medical school educators: an untapped resource. AB - Despite extensive experience teaching residents, surgeons are an untapped resource for educating medical students. We hypothesized that by involving surgeons as teachers earlier in the medical school curriculum, medical students' interest in surgery will increase and their opinions of surgeons will improve. Five programs designed to involve surgeons as educators in the medical school curriculum were implemented. The first program, started in 2008, introduced surgical faculty into the first-year medical student anatomy dissection laboratories. Other programs initiated in 2008 included: Surgical Clinical Correlates in Anatomy, which involved faculty teaching through cadaver surgery; Clinical Pathologic Conferences in Anatomy, a surgeon-led conference based on clinical cases; and a women's faculty-student mentorship program. Table Rounds, a surgeon-led anatomy review that used clinical scenarios to quiz students was begun in 2009. All five programs were successfully integrated into the medical school curriculum. While student opinion of surgeons as educators improved, there were no significant changes in student interest in surgery as a career nor change in performance on written examinations over the Anatomy content covered by the surgeons. Surgical faculty and trainees can be integrated into the medical school curriculum. Involving surgeons as educators earlier in the medical school curriculum may have longer term effects than could be observed in this study. At a minimum, the experience improved student opinion of surgeons as educators. PMID- 21618447 TI - Medical students' perceptions of the body donor as a "first patient" or "teacher": a pilot study. AB - University of Michigan Medical School (UMMS) students attending a seminar on the history and ethics of anatomical dissection were fascinated by a report on the dissection room experience in Thailand that relates the body donor's status as a teacher. The students felt that they had naturally adopted the "body as teacher" approach in their dissection course, rather than the "body as first patient" approach that is encouraged by faculty. It was decided to explore the question whether other medical students shared these perceptions. A questionnaire was sent out to all UMMS students who had finished the anatomical dissection course. One hundred twenty-eight responses from a population of 500 students were received. Results indicate that students believe the "body as teacher" approach is more effective in engendering respect and empathy towards the body and towards future patients, and in facilitating students' emotional development. Students also reported wanting a more personal relationship with their donors. Eighty four percent of students preferred the "body as teacher" approach to the currently taught "body as first patient" approach. The results support the hypothesis that students' desired closer personal relationship with donors might be better facilitated by the "body as teacher" approach, and that this closer relationship engenders empathy and respect towards the donor and future patients. A new model for anatomy programs could introduce the donor first as a teacher and later transition into viewing the donor as a patient. PMID- 21618448 TI - Nanoparticles through the skin: managing conflicting results of inorganic and organic particles in cosmetics and pharmaceutics. AB - Toxicity of nanoparticles is a current scientific issue because of the enhanced reactivity of nanomaterials and their possible easy penetration into the body arising from their small size. Because inorganic particles are present in sunscreen cosmetic products, attention has been focused on cutaneous penetration. But organic particles of various sizes are also used in pharmaceutical applications such as skin care and transdermal drug delivery. It appears that organic and inorganic particles penetrate the skin quite differently. The apparent discrepancy is addressed in this review focusing on skin penetration of inorganic sunscreen particles and organic particles for drug delivery. After a short description of the physicochemical properties of these particles, the skin penetration of both types is reviewed with emphasis on the mechanistic issues and the differences that could account for such conflicting results. It appears that investigations by cosmetic and pharmaceutical communities focused on the main issue, i.e., no toxicity in cosmetics and maximum activity of the drug in pharmaceutics. This leaves several fundamental issues as open questions and this does not allow a rigorous comparison between both types of material. While it is claimed that inorganic nanoparticles can only penetrate the outer layer of the skin, it appears that organic submicron particles and even microparticles reach the dermis in an in vitro cell. Besides particle size, the surface chemistry of the particles and the presence of other excipients in the formulations contribute to skin absorption. PMID- 21618449 TI - Unraveling DNA dynamics using atomic force microscopy. AB - The elucidation of structure-function relationships of biological samples has become important issue in post-genomic researches. In order to unveil the molecular mechanisms controlling gene regulations, it is essential to understand the interplay between fundamental DNA properties and the dynamics of the entire molecule. The wide range of applicability of atomic force microscopy (AFM) has allowed us to extract physicochemical properties of DNA and DNA-protein complexes, as well as to determine their topographical information. Here, we review how AFM techniques have been utilized to study DNA and DNA-protein complexes and what types of analyses have accelerated the understanding of the DNA dynamics. We begin by illustrating the application of AFM to investigate the fundamental feature of DNA molecules; topological transition of DNA, length dependent properties of DNA molecules, flexibility of double-stranded DNA, and capability of the formation of non-Watson-Crick base pairing. These properties of DNA are critical for the DNA folding and enzymatic reactions. The technical advancement in the time-resolution of AFM and sample preparation methods enabled visual analysis of DNA-protein interactions at sub-second time region. DNA tension-dependent enzymatic reaction and DNA looping dynamics by restriction enzymes were examined at a nanoscale in physiological environments. Contribution of physical properties of DNA to dynamics of nucleosomes and transition of the higher-order structure of reconstituted chromatin are also reviewed. PMID- 21618450 TI - Diagnostic accuracy, internal consistency, and convergent validity of the Greek version of the patient health questionnaire 9 in diagnosing depression in rheumatologic disorders. AB - OBJECTIVE: The Patient Health Questionnaire 9 (PHQ-9) was developed to screen for depressive disorders in community, primary care, and medical settings. We aimed to estimate its diagnostic accuracy, internal consistency, reliability, and convergent validity in diagnosing major depressive disorder (MDD) in Greek patients with rheumatologic disorders. METHODS: In a 2-phase sampling design study, we recruited 475 patients with established rheumatologic disorders. One of 2 of the high scorers (PHQ-9 score >=9, n = 85) and 1 of 3 of the low scorers (PHQ-9 score 0-8, n = 128) were interviewed using the Mini International Neuropsychiatric Interview to confirm MDD. A receiver operator characteristic curve analysis was performed to confirm the optimum threshold value. The scale's dimensional structure was tested with factor analysis, and internal consistency reliability was assessed with Cronbach's alpha. Psychological distress (Symptom Check List-90-Revised [SCL-90-R]), disability (Health Assessment Questionnaire disability index), and health-related quality of life (HRQOL; World Health Organization Quality of Life Instrument [WHOQOL-BREF]) were also assessed to test convergent validity with bivariate correlations. RESULTS: At an optimum threshold of 10, the PHQ-9 showed a sensitivity of 81.2% and a specificity of 86.8%. The area under the curve was 0.91. The PHQ-9 presented unidimensional structure with good scale reliability (alpha = 0.82). The PHQ-9 score presented the greatest correlations with SCL-90-R depression (r = 0.736) and WHOQOL-BREF mental HRQOL scales (r = -0.571), and all other correlations with disability and HRQOL were in the expected direction. CONCLUSION: At a cutoff of 10, the PHQ-9 is an accurate, reliable, and valid measure for screening for MDD among Greek rheumatologic patients. PMID- 21618451 TI - Association of the Charlson comorbidity index with mortality in systemic lupus erythematosus. AB - OBJECTIVE: To investigate whether comorbidity as assessed by the Charlson Comorbidity Index (CCI) is associated with mortality in a long-term followup of systemic lupus erythematosus (SLE) patients. METHODS: Data were collected from 499 SLE patients attending the Lupus Clinic at the McGill University Health Center, Montreal, Quebec, Canada, and 170 SLE patients from the Department of Rheumatology at Lund University Hospital, Lund, Sweden. This included data on comorbidity, demographics, disease activity, the Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index (SDI), and antiphospholipid antibody syndrome (APS). Variables were entered into a Cox proportional hazards survival model. RESULTS: Mortality risk in the Montreal cohort was associated with the CCI (hazard ratio [HR] 1.57 per unit increase in the CCI, 95% confidence interval [95% CI] 1.18-2.09) and age (HR 1.04 per year increase in age, 95% CI 1.00-1.09). The CCI and age at diagnosis were also associated with mortality in the Lund cohort (CCI: HR 1.35, 95% CI 1.13-1.60; age: HR 1.09, 95% CI 1.05-1.12). Furthermore, the SDI was associated with mortality in the Lund cohort (HR 1.40, 95% CI 1.19-1.64), while a wide CI for the estimate in the Montreal cohort prevented a definitive conclusion (HR 1.20, 95% CI 0.97-1.48). We did not find a strong association between mortality and sex, race/ethnicity, disease activity, or APS in either cohort. CONCLUSION: In this study, comorbidity as measured by the CCI was associated with decreased survival independent of age, lupus disease activity, and damage. This suggests that the CCI may be useful in capturing comorbidity for clinical research in SLE. PMID- 21618453 TI - Enthesitis in an inception cohort of enthesitis-related arthritis. AB - OBJECTIVE: To describe an enthesitis-related arthritis (ERA) inception cohort and determine which entheses and joints are most commonly affected. METHODS: We reviewed a retrospective inception cohort study of children with ERA who were diagnosed and treated at The Children's Hospital of Philadelphia between November 2007 and December 2009. RESULTS: During the study period, there were 32 newly diagnosed ERA patients. Fifty-nine percent were male, and the median age at the date of initial evaluation was 12.5 years (interquartile range [IQR] 10.2-14.3 years). The median number of tender entheses at presentation was 2 (IQR 0-5), and 21 subjects (66%) had at least 1 tender enthesis. The most prevalent tender entheses were the patellar ligament insertion at the inferior pole of the patella, the plantar fascial insertion at the calcaneus, the Achilles tendon insertion at the calcaneus, and the plantar fascial insertion at the metatarsal heads. Enthesitis was most often symmetric. The median number of active joints was 2 (IQR 0-4). The most commonly affected joints were the sacroiliacs, knees, and ankles. Sacroiliitis, which was defined clinically, was most often symmetric, while peripheral arthritis was most frequently asymmetric. The odds of having active enthesitis at 6 months increased significantly with each additional tender enthesis at the initial evaluation. CONCLUSION: Among pediatric patients with ERA, lower extremity enthesitis is prevalent at the time of diagnosis and is likely to persist 6 months later. Future studies should address standardization of the enthesitis examination, the pattern of enthesitis over time, enthesitis response to therapy, and the impact of enthesitis on quality of life. PMID- 21618452 TI - Preliminary criteria for global flares in childhood-onset systemic lupus erythematosus. AB - OBJECTIVE: To develop widely acceptable preliminary criteria of global flare for childhood-onset systemic lupus erythematosus (cSLE). METHODS: Pediatric rheumatologists (n = 138) rated a total of 358 unique patient profiles with information about the cSLE flare descriptors from 2 consecutive visits: patient global assessment of well-being, physician global assessment of disease activity (MD-global), health-related quality of life, anti-double-stranded DNA antibodies, disease activity index scores, protein:creatinine (P:C) ratio, complement levels, and erythrocyte sedimentation rate (ESR). Based on 2,996 rater responses about the course of cSLE (baseline versus followup), the accuracy (sensitivity, specificity, and area under the receiver operating characteristic curve) of candidate flare criteria was assessed. An international consensus conference was held to rank these candidate flare criteria as per the American College of Rheumatology recommendations for the development and validation of criteria sets. RESULTS: The highest-ranked candidate criteria considered absolute changes (Delta) of the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) or British Isles Lupus Assessment Group (BILAG), MD-global, P:C ratio, and ESR; flare scores can be calculated (0.5 * DeltaSLEDAI + 0.45 * DeltaP:C ratio + 0.5 * DeltaMD-global + 0.02 * DeltaESR), where values of >=1.04 are reflective of a flare. Similarly, BILAG-based flare scores (0.4 * DeltaBILAG + 0.65 * DeltaP:C ratio + 0.5 * DeltaMD-global + 0.02 * DeltaESR) of >=1.15 were diagnostic of a flare. Flare scores increased with flare severity. CONCLUSION: Consensus has been reached on preliminary criteria for global flares in cSLE. Further validation studies are needed to confirm the usefulness of the cSLE flare criteria in research and for clinical care. PMID- 21618454 TI - Development and initial validation of composite parent- and child-centered disease assessment indices for juvenile idiopathic arthritis. AB - OBJECTIVE: To develop and validate a parent-centered and a child-centered composite disease assessment index for juvenile idiopathic arthritis (JIA): the Juvenile Arthritis Parent Assessment Index (JAPAI) and the Juvenile Arthritis Child Assessment Index (JACAI), respectively. METHODS: The JAPAI and the JACAI include 4 measures: parent/child rating of overall well-being, pain, physical function, and health-related quality of life (HRQOL). Validation analyses were conducted on nearly 5,000 patients and included assessment of construct validity, discriminant validity, responsiveness to change, and reliability. Besides the 4 item version, a 3-item version of both indices, which did not include HRQOL, was tested. RESULTS: The JAPAI and the JACAI demonstrated good construct validity, yielding high correlations with the Juvenile Arthritis Disease Activity Score and moderate correlations with physician global rating and joint counts. Correlations obtained for the JAPAI and the JACAI and for the 4-item and the 3-item versions were comparable. Factorial analysis by principal component analysis showed that both indices are monodimensional. Both the JAPAI and JACAI discriminated well between different disease states and courses and between different levels of American College of Rheumatology Pediatric criteria in a clinical trial, and revealed fair responsiveness to clinical change. Internal consistency was satisfactory, with a Cronbach's alpha of >0.80 in all but 1 of the patient samples tested. CONCLUSION: The JAPAI and the JACAI were found to be valid instruments for assessment of disease status in JIA and suitable surrogates of physicians' evaluations. Both indices are potentially applicable in clinical practice, observational studies, and therapeutic trials. PMID- 21618455 TI - Luteinized unruptured follicle syndrome increased by inactive disease and selective cyclooxygenase 2 inhibitors in women with inflammatory arthropathies. AB - OBJECTIVE: Administration of nonsteroidal antiinflammatory drugs (NSAIDs) may impair fertility. The occurrence of the luteinized unruptured follicle (LUF) syndrome was assessed in women with inflammatory arthropathies exposed to NSAIDs and compared to that in nonexposed women. METHODS: Fourteen patients with inflammatory rheumatic disease, 29 women with noninflammatory musculoskeletal conditions, and 449 women not exposed to NSAIDs were studied by intravaginal ultrasound monitoring for follicular development and ovulation in 1 or more menstrual cycles. Disease activity was assessed in inflammatory rheumatic disease. RESULTS: In 59 monitored cycles of patients with continuous NSAID exposure, 35.6% of LUF syndromes occurred compared to 3.4% of LUF syndromes in untreated women (P < 0.001). Etoricoxib was responsible for 75% of LUF syndromes in patients exposed continuously, whereas diclofenac generated 15% of LUF syndromes. An ibuprofen dosage of 1,600 mg/day did not induce LUF syndrome either at continuous periovulatory or discontinuous exposure. Interestingly, the frequency of LUF syndrome was 46.2% in patients with inactive inflammatory disease compared to 15% in patients with active disease (P = 0.023). Etoricoxib generated LUF syndrome in 94.2% of the cases with inactive disease versus 28.6% in patients with active disease (P = 0.003). CONCLUSION: NSAIDs increased the risk of the LUF syndrome, particularly in patients with inactive disease. The selective cyclooxygenase 2 (COX-2) inhibitor etoricoxib was a more potent inductor of LUF syndrome than nonselective COX inhibitors. Continuous periovulatory exposure to NSAIDs should be avoided when planning a pregnancy in patients with rheumatic diseases. PMID- 21618456 TI - Integrated approach to produce a recombinant, His-tagged human alpha galactosidase A in mammalian cells. AB - Successful production of recombinant proteins (r-proteins) by transient gene expression (TGE) depends on several parameters (including producer cells, culture conditions, transfection procedure, or expression vector) that should be optimized when producing any recombinant product. In this work, TGE-based production of human alpha-galactosidase A (GLA) is described. Producer cells, expression vectors, and parameters influencing cell metabolism after transfection have been tested. The enzyme is secreted, has the right molecular weight, and is enzymatically active. Productivities of up to 30-40 mg/L have been achieved, with a simple, fast procedure. A 6 * His tag allows enzyme purification in a single step, rendering a highly pure product. We propose a TGE-based protocol able to produce up to several milligrams per liter of highly pure, active GLA in a time as short as a few days. By this, enough amounts of engineered versions of the enzyme can be easily produced to be tested in vitro or in preclinical trials. PMID- 21618457 TI - Assembly of horseradish peroxidase within supported cationic bilayers. AB - The interaction between horseradish peroxidase (HRP) and dioctadecyldimethylammonium bromide (DODAB) bilayers supported on polystyrene microspheres (PSS) or on flat silicon wafers was evaluated from the following techniques: (1) dynamic light-scattering for determining size distributions, zeta potentials and polydispersities for dispersions; (2) spectrophotometric determination of HRP concentration in supernatants of centrifuged mixtures; (3) in situ ellipsometry for mean thickness of deposited layers on wafers; (4) kinetics of product appearance for oxidation of 2,2'-azino-bis-3 ethylbenzothiazoline-6-sulfonic acid by H(2) O(2) in presence of free or immobilized enzyme. HRP incorporation (3.0 mg/m(2)) did not alter mean diameter and zeta-potential of PSS/DODAB particles but reduced enzyme activity by 50%, though activity persisted after several rinsing steps. In situ ellipsometry could not detect any HRP layer on top of the DODAB bilayer. HRP insertion in the bilayer core explained all results for both systems. Useful biotechnological applications are anticipated for such assemblies. PMID- 21618458 TI - Development of a mathematical model for evaluating the dynamics of normal and apoptotic Chinese hamster ovary cells. AB - A metabolic flux based methodology was developed for modeling the metabolism of a Chinese hamster ovary cell line. The elimination of insignificant fluxes resulted in a simplified metabolic network which was the basis for modeling the significant metabolites. Employing kinetic rate expressions for growing and non growing subpopulations, a logistic model was developed for cell growth and dynamic models were formulated to describe culture composition and monoclonal antibody (MAb) secretion. The model was validated for a range of nutrient concentrations. Good agreement was obtained between model predictions and experimental data. The ultimate goal of this study is to establish a comprehensive dynamic model which may be used for model-based optimization of the cell culture for MAb production in both batch and fed-batch systems. PMID- 21618459 TI - Novel assays of thrombogenic pathogenicity in the antiphospholipid syndrome based on the detection of molecular oxidative modification of the major autoantigen beta2-glycoprotein I. AB - OBJECTIVE: Beta-2-glycoprotein I (beta2 GPI) constitutes the major autoantigen in the antiphospholipid syndrome (APS), a common acquired cause of arterial and venous thrombosis. We recently described the novel observation that beta2 GPI may exist in healthy individuals in a free thiol (biochemically reduced) form. The present study was undertaken to quantify the levels of total, reduced, and posttranslationally modified oxidized beta2 GPI in APS patients compared to various control groups. METHODS: In a retrospective multicenter analysis, the proportion of beta2 GPI with free thiols in serum from healthy volunteers was quantified. Assays for measurement of reduced as well as total circulating beta2 GPI were developed and tested in the following groups: APS (with thrombosis) (n=139), autoimmune disease with or without persistent antiphospholipid antibodies (aPL) but without APS (n=188), vascular thrombosis without APS or aPL (n=38), and healthy volunteers (n=91). RESULTS: Total beta2 GPI was significantly elevated in patients with APS (median 216.2 MUg/ml [interquartile range 173.3 263.8]) as compared to healthy subjects (median 178.4 MUg/ml [interquartile range 149.4-227.5] [P<0.0002]) or control patients with autoimmune disease or vascular thrombosis (both P<0.0001). The proportion of total beta2 GPI in an oxidized form (i.e., lacking free thiols) was significantly greater in the APS group than in each of the 3 control groups (all P<0.0001). CONCLUSION: This large retrospective multicenter study shows that posttranslational modification of beta2 GPI via thiol-exchange reactions is a highly specific phenomenon in the setting of APS thrombosis. Quantification of posttranslational modifications of beta2 GPI in conjunction with standard laboratory tests for APS may offer the potential to more accurately predict the risk of occurrence of a thrombotic event in the setting of APS. PMID- 21618460 TI - Neuroimaging evidence of white matter inflammation in newly diagnosed systemic lupus erythematosus. AB - OBJECTIVE: Central nervous system (CNS) involvement occurs frequently in systemic lupus erythematosus (SLE) and frequently results in morbidity. The primary pathophysiology of CNS involvement in SLE is thought to be inflammation secondary to autoantibody-mediated vasculitis. Neuroimaging studies have shown hypometabolism (representing impending cell failure) and atrophy (representing late-stage pathology), but not inflammation. The purpose of this study was to detect the presence and regional distribution of inflammation (hypermetabolism) and tissue failure, apoptosis, or atrophy (hypometabolism). METHODS: Eighty-five patients with newly diagnosed SLE, who had no focal neurologic symptoms, were studied. Disease activity was quantified using the Safety of Estrogens in Lupus Erythematosus: National Assessment version of the SLE Disease Activity Index (SELENA-SLEDAI), a validated index of SLE-related disease activity. 18Fluorodeoxyglucose (FDG) positron emission tomography (PET) images of glucose uptake were analyzed by visual inspection and as group statistical parametric images, using the SELENA-SLEDAI score as the analysis regressor. RESULTS: SELENA SLEDAI-correlated increases in glucose uptake were found throughout the white matter, most markedly in heavily myelinated tracts. SELENA-SLEDAI-correlated decreases were found in the frontal and parietal cortex, in a pattern similar to that seen during visual inspection and presented in previous reports of hypometabolism. CONCLUSION: The SELENA-SLEDAI-correlated increases in glucose consumption are potential evidence of inflammation, consistent with prior reports of hypermetabolism in inflammatory disorders. To our knowledge, this is the first imaging-based evidence of SLE-induced CNS inflammation in an SLE inception cohort. The dissociation among 18FDG uptake characteristics, spatial distribution, and disease activity correlation is in accordance with the notion that glucose hypermetabolism and hypometabolism reflect fundamentally different aspects of the pathophysiology of SLE with CNS involvement. PMID- 21618461 TI - Folate-targeted nanoparticles show efficacy in the treatment of inflammatory arthritis. AB - OBJECTIVE: To investigate the uptake of a poly(amidoamine) dendrimer (generation 5 [G5]) nanoparticle covalently conjugated to polyvalent folic acid (FA) as the targeting ligand into macrophages, and to investigate the activity of an FA- and methotrexate (MTX)-conjugated dendrimer (G5-FA-MTX) as a therapeutic for the inflammatory disease of arthritis. METHODS: In vitro studies were performed in macrophage cell lines and in isolated mouse macrophages to check the cellular uptake of fluorescence-tagged G5-FA nanoparticles, using flow cytometry and confocal microscopy. In vivo studies were conducted in a rat model of collagen induced arthritis to evaluate the therapeutic potential of G5-FA-MTX. RESULTS: Folate-targeted dendrimer bound and internalized in a receptor-specific manner into both folate receptor beta-expressing macrophage cell lines and primary mouse macrophages. The conjugate G5-FA-MTX acted as a potent antiinflammatory agent and reduced arthritis-induced parameters of inflammation such as ankle swelling, paw volume, cartilage damage, bone resorption, and body weight decrease. CONCLUSION: The use of folate-targeted nanoparticles to specifically target MTX into macrophages may provide an effective clinical approach for antiinflammatory therapy in rheumatoid arthritis. PMID- 21618462 TI - Blockade of NKG2D ameliorates disease in mice with collagen-induced arthritis: a potential pathogenic role in chronic inflammatory arthritis. AB - OBJECTIVE: To assess the role of the activating receptor NKG2D in arthritis. METHODS: Levels of NKG2D and its ligands were determined by fluorescence activated cell sorting, real-time polymerase chain reaction, and immunohistochemistry in rheumatoid arthritis (RA) synovial membrane tissue and in paw tissue from arthritic mice. Arthritis was induced in DBA/1 mice by immunization with type II collagen, and mice were treated intraperitoneally with a blocking anti-NKG2D antibody (CX5) on days 1, 5, and 8 after clinical onset and were monitored for 10 days. RESULTS: We demonstrated expression of NKG2D and its ligands on human RA synovial cells and extended this finding to the paws of arthritic mice. Expression of messenger RNA for the NKG2D ligand Rae-1 was up regulated, and NKG2D was present predominantly on natural killer (NK) and CD4+ T cells, in arthritic paw cell isolates. NKG2D was down-modulated during the progression of collagen-induced arthritis (CIA). NKG2D expression in arthritic paws was demonstrated by immunohistochemistry. Blockade of NKG2D ameliorated established CIA, with significant reductions in clinical scores and paw swelling. Histologic analysis of arthritic joints from anti-NKG2D-treated mice demonstrated significant joint protection, compared with control mice. Moreover, anti-NKG2D treatment significantly reduced both interleukin-17 production from CD4+ T cells in arthritic paws and splenic NK cell cytotoxic effector functions in vivo and in vitro. CONCLUSION: Our findings indicate that blockade of NKG2D in a murine model and in human explants has beneficial therapeutic potential that merits further investigation in RA. PMID- 21618464 TI - New era of molecular structure and dynamics from solution scattering experiments. PMID- 21618465 TI - Ammonia-oxidizing bacteria dominates over ammonia-oxidizing archaea in a saline nitrification reactor under low DO and high nitrogen loading. AB - A continuous nitrification reactor treating saline wastewater was operated for almost 1 year under low dissolved oxygen (DO) levels (0.15-0.5 mg/L) and high nitrogen loadings (0.26-0.52 kg-N/(m(3) day)) in four phases. The diversity and abundance of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) were analyzed by cloning, terminal restriction fragment length polymorphism (T RFLP) and quantitative polymerase chain reaction (qPCR). The results showed that there were only one dominant AOA species and one dominant AOB species in the reactor in all of the four experimental phases. The amoA gene of the dominant AOA only had a similarity of 89.3% with the cultured AOA species Nitrosopumilus maritimus SCM1. All of the AOB species detected in the reactor belong to Nitrosomonas genus and it was found that the AOB populations changed with the ammonium loadings and DO levels. The abundance of AOB in the reactor was ~40 times larger than that of AOA, and the ratio of AOB to AOA increased significantly up to ~2,000 to ~4,000 with the increase of ammonium loading, indicating that AOB are much more competitive than AOA in high ammonium environments and probably AOA play a less important role than AOB in the nitrification reactors. PMID- 21618466 TI - Comparative metabolic capabilities for Micrococcus luteus NCTC 2665, the "Fleming" strain, and actinobacteria. AB - Putative gene predictions of the Gram positive actinobacteria Micrococcus luteus (NCTC 2665, "Fleming strain") was used to construct a genome scale reconstruction of the metabolic network for this organism. The metabolic network comprises 586 reactions and 551 metabolites, and accounts for 21% of the genes in the genome. The reconstruction was based on the annotated genome and available biochemical information. M. luteus has one of the smallest genomes of actinobacteria with a circular chromosome of 2,501,097 base pairs and a GC content of 73%. The metabolic pathways required for biomass production in silico were determined based on earlier models of actinobacteria. The in silico network is used for metabolic comparison of M. luteus with other actinomycetes, and hence provides useful information for possible future biotechnological exploitation of this organism, e.g., for production of biofuels. PMID- 21618467 TI - Selective detection of gold using genetically engineered bacterial reporters. AB - Salmonella typhimurium harbours a Au-resistance system whose expression is controlled by GolS, a transcriptional regulator of the MerR family that selectively detects Au with high sensitivity. We developed both Salmonella and genetically engineered Escherichia coli strains as Au-selective whole-cell biosensors by coupling the strictly regulated GolS-dependent golB promoter to the gfp reporter gene. The bio-reporters were evaluated under different laboratory conditions and calibrated for their use as selective Au detectors. Due to the intrinsic characteristics of the regulatory protein, the transgenic E. coli sensor exhibits low background, high signal-to-noise ratio, and improved sensitivity for detection of Au ions in a wide range of concentrations (up to 470 nM) with a calculated detection limit of ~33 nM (6 ug L(-1) or parts per billion) Au(I). The fluorescent Au-sensing bacteria exhibit also minimal interference by chemically related metals such as Cu or Ag that are commonly found in Au deposits. These highly specific and sensitive Au detectors might allow the development of rapid and robust screening tools to improve discovery and extraction procedures. PMID- 21618468 TI - Developing a compound-specific receptor for bisphenol A by directed evolution of human estrogen receptor alpha. AB - Directed evolution has become a successful approach to alter ligand binding properties of nuclear receptors. In this study, directed evolution was used to generate a mutant human estrogen receptor alpha library, which was then used to screen for receptors having enhanced responses to the known endocrine-disrupting chemical, bisphenol A (BPA). A single round of multi-site mutation was combined with an efficient positive/negative library screening method in which positive growth-based selection for the desired activity with BPA was combined with flow cytometric removal of cells having undesired activity with the natural ligand, 17beta-estradiol. The screening steps were performed in a Saccharomyces cerevisiae yeast strain containing a genome-integrated his3-yEGFP reporter gene fusion construct. A single round of mutation and screening yielded nine mutants with enhanced responses towards BPA but no detectable induction by 17beta estradiol (up to 90 nM). These BPA-specific mutant receptors may prove useful in the field of environmental analytics, where they could be used to monitor and evaluate the proportion of BPA in hormonally active samples. PMID- 21618469 TI - Metabolic adaptation of MDCK cells to different growth conditions: effects on catalytic activities of central metabolic enzymes. AB - Lactate and ammonia are the most important waste products of central carbon metabolism in mammalian cell cultures. In particular during batch and fed-batch cultivations these toxic by-products are excreted into the medium in large amounts, and not only affect cell viability and productivity but often also prevent growth to high cell densities. The most promising approach to overcome such a metabolic imbalance is the replacement of one or several components in the culture medium. It has been previously shown that pyruvate can be substituted for glutamine in cultures of adherent Madin-Darby canine kidney (MDCK) cells. As a consequence, the cells not only released no ammonia but glucose consumption and lactate production were also reduced significantly. In this work, the impact of media changes on glucose and glutamine metabolism was further elucidated by using a high-throughput platform for enzyme activity measurements of mammalian cells. Adherent MDCK cells were grown to stationary and exponential phase in six-well plates in serum-containing GMEM supplemented with glutamine or pyruvate. A total number of 28 key metabolic enzyme activities of cell extracts were analyzed. The overall activity of the pentose phosphate pathway was up-regulated during exponential cell growth in pyruvate-containing medium suggesting that more glucose-6-phosphate was channeled into the oxidative branch. Furthermore, the anaplerotic enzymes pyruvate carboxylase and pyruvate dehydrogenase showed higher cell specific activities with pyruvate. An increase in cell specific activity was also found for NAD(+)-dependent isocitrate dehydrogenase, glutamate dehydrogenase, and glutamine synthetase in MDCK cells grown with pyruvate. It can be assumed that the increase in enzyme activities was required to compensate for the energy demand and to replenish the glutamine pool. On the other hand, the activities of glutaminolytic enzymes (e.g., alanine and aspartate transaminase) were decreased in cells grown with pyruvate, which seems to be related to a decreased glutamine metabolism. PMID- 21618470 TI - Promoter methylation and transgene copy numbers predict unstable protein production in recombinant Chinese hamster ovary cell lines. AB - Mammalian cell lines for recombinant protein production need to maintain productivity over extended cultivation times. Long-term stability studies are time and resource intensive, but are widely performed to identify and eliminate unstable candidates during cell line development. Production instability of manufacturing cell lines can be associated with methylation and silencing of the heterologous promoter. We have identified CpG dinucleotides within the human cytomegalovirus major immediate early promoter/enhancer (hCMV-MIE) that are frequently methylated in unstable antibody-producing Chinese hamster ovary (CHO) cell lines. We have established methylation-specific real-time qPCR for the rapid and sensitive measurement of hCMV-MIE methylation in multiple cell lines and provide evidence that hCMV-MIE methylation and transgene copy numbers can be used as early markers to predict production instability of recombinant CHO cell lines. These markers should provide the opportunity to enrich stable producers early in cell line development and allow developers to put more emphasis on other criteria, such as product quality and bioprocess robustness. PMID- 21618471 TI - Transforming growth factor-beta1 is constitutively secreted by Chinese hamster ovary cells and is functional in human cells. AB - Chinese hamster ovary (CHO) cells are widely used for the production of recombinant proteins for clinical use as well as academic research. They are particularly important for the production of glycoproteins where bacteria cannot be used. TGFbeta1 is a potent cytokine highly conserved across species with multiple immunological and non-immunological effects. We have discovered that CHOK1, the CHO clone most commonly used by the pharmaceutical industry, constitutively secretes latent TGFbeta1 and that this hamster TGFbeta1 is active on human cells inducing profound immunological effects. As far as we are aware, the production of TGFbeta1 by CHOK1 cells has not been reported before in the literature. As TGFbeta1 exerts powerful and pleiotropic effects on diverse cell types, and as CHO cells are used to produce a large number of clinical and non clinical products, our findings are highly relevant to studies that rely on recombinant proteins. PMID- 21618472 TI - Controlling glycation of recombinant antibody in fed-batch cell cultures. AB - Protein glycation is a non-enzymatic glycosylation that can occur to proteins in the human body, and it is implicated in the pathogenesis of multiple chronic diseases. Glycation can also occur to recombinant antibodies during cell culture, which generates structural heterogeneity in the product. In a previous study, we discovered unusually high levels of glycation (>50%) in a recombinant monoclonal antibody (rhuMAb) produced by CHO cells. Prior to that discovery, we had not encountered such high levels of glycation in other in-house therapeutic antibodies. Our goal here is to develop cell culture strategies to decrease rhuMAb glycation in a reliable, reproducible, and scalable manner. Because glycation is a post-translational chemical reaction between a reducing sugar and a protein amine group, we hypothesized that lowering the concentration of glucose -the only source of reducing sugar in our fed-batch cultures--would lower the extent of rhuMAb glycation. When we decreased the supply of glucose to bioreactors from bolus nutrient and glucose feeds, rhuMAb glycation decreased to below 20% at both 2-L and 400-L scales. When we maintained glucose concentrations at lower levels in bioreactors with continuous feeds, we could further decrease rhuMAb glycation levels to below 10%. These results show that we can control glycation of secreted proteins by controlling the glucose concentration in the cell culture. In addition, our data suggest that rhuMAb glycation occurring during the cell culture process may be approximated as a second-order chemical reaction that is first order with respect to both glucose and non-glycated rhuMAb. The basic principles of this glycation model should apply to other recombinant proteins secreted during cell culture. PMID- 21618473 TI - Utilization of non-AUG initiation codons in a flow cytometric method for efficient selection of recombinant cell lines. AB - Here we describe a method that couples flow cytometric detection with the attenuated translation of a reporter protein to enable efficient selection of CHO clones producing high levels of recombinant proteins. In this system, a small cell surface reporter protein is expressed from an upstream open reading frame utilizing a non-AUG initiation (alternate start) codon. Due to the low translation initiation efficiency of this alternate start codon, the majority of translation initiation events occur at the first AUG of the downstream open reading frame encoding the recombinant protein of interest. While translation of the reporter is significantly reduced, the levels are sufficient for detection using flow cytometric methods and, in turn, predictive of protein expression from the gene of interest since both ORFs are translated from the same mRNA. Using this system, CHO cells have been sorted to obtain enriched pools producing significantly higher levels of recombinant proteins than the starting cell population and clones with significantly better productivity than those generated from limiting dilution cloning. This method also serves as an effective screening tool during clone expansion to enable resources to be focused solely on clones with both high and stable expression. PMID- 21618474 TI - Evaluation of protein engineering and process optimization approaches to enhance antibody drug manufacturability. AB - A potent single digit picomolar fully human monoclonal antibody (hMAb) inhibitor with a high degree of specificity to the antigen of interest was identified from a phage display library. The hMAb, however, exhibited a high degree of hydrophobicity and easily formed insoluble aggregates when purified using a Protein A based generic process. Strategies were designed using both protein engineering and process development approaches to optimize the molecule's amino acid sequence and its behavior in process conditions. The insoluble aggregation issue was brought under control by one single amino acid mutation in CDR region or by switching to non-ProA based purification process. Our study therefore presents the rational manufacturability design for future monoclonal antibody product and its purification process under the quality by design concept by either engineering the drug molecule to adapt existing platform process or optimizing the process to fit the specific properties of the drug product. PMID- 21618475 TI - The role of polymer nanolayer architecture on the separation performance of anion exchange membrane adsorbers: I. Protein separations. AB - This contribution describes the preparation of strong anion-exchange membranes with higher protein binding capacities than the best commercial resins. Quaternary amine (Q-type) anion-exchange membranes were prepared by grafting polyelectrolyte nanolayers from the surfaces of macroporous membrane supports. A focus of this study was to better understand the role of polymer nanolayer architecture on protein binding. Membranes were prepared with different polymer chain graft densities using a newly developed surface-initiated polymerization protocol designed to provide uniform and variable chain spacing. Bovine serum albumin and immunoglobulin G were used to measure binding capacities of proteins with different size. Dynamic binding capacities of IgG were measured to evaluate the impact of polymer chain density on the accessibility of large size protein to binding sites within the polyelectrolyte nanolayer under flow conditions. The dynamic binding capacity of IgG increased nearly linearly with increasing polymer chain density, which suggests that the spacing between polymer chains is sufficient for IgG to access binding sites all along the grafted polymer chains. Furthermore, the high dynamic binding capacity of IgG (>130 mg/mL) was independent of linear flow velocity, which suggests that the mass transfer of IgG molecules to the binding sites occurs primarily via convection. Overall, this research provides clear evidence that the dynamic binding capacities of large biologics can be higher for well-designed macroporous membrane adsorbers than commercial membrane or resin ion-exchange products. Specifically, using controlled polymerization leads to anion-exchange membrane adsorbers with high binding capacities that are independent of flow rate, enabling high throughput. Results of this work should help to accelerate the broader implementation of membrane adsorbers in bioprocess purification steps. PMID- 21618476 TI - The role of polymer nanolayer architecture on the separation performance of anion exchange membrane adsorbers: part II. DNA and virus separations. AB - The surface-initiated polymerization protocol developed in part I was used to prepare strong anion-exchange membranes with variable polymer chain graft densities and degrees of polymerization for DNA and virus particle separations. A focus of part II was to evaluate the role of polymer nanolayer architecture on DNA and virus binding. Salmon sperm-DNA (SS-DNA) was used as model nucleic acid to measure the dynamic-binding capacities at 10% breakthrough. The dynamic binding capacity increases linearly with increasing poly ([2 (methacryloyloxy)ethyl]trimethylammonium chloride) chain density up to the highest chain density used in this study. The new membranes yielded threefold higher SS-DNA-binding capacity (30 mg/mL) than a leading commercial membrane with the same functional group chemistry. Elution of bound DNA yielded a sharp peak, and resulted in a 13-fold increase relative to the feed concentration. This concentration effect further demonstrates the highly favorable transport properties of the newly designed Q-type membranes. However, unlike findings in part I on protein binding, SS-DNA binding was not fully reversible. Minute virus of mice (MVM) was used as model virus to evaluate the virus clearance performance of newly designed Q-type membranes. Log reduction of virus (LRV) of MVM increased with increasing polymer chain density. Membranes exhibited >4.5 LRV for the given MVM impurity load and may be capable of higher LRV values, as the MVM concentration in the flow-through fraction of these samples was below the limit of detection of the assay. PMID- 21618477 TI - Autophagy induced by rapamycin and carbon-starvation have distinct proteome profiles in Aspergillus nidulans. AB - It is hypothesized that autophagy, a global catabolic pathway which is highly conserved from yeast to man, plays an important role in many bioprocesses. Though autophagy is known to be induced by either nutrient starvation or treatment with the drug rapamycin, it is not clear whether the two modes of induction have the same long-term impact in the cell, particularly in the biotechnologically important filamentous fungi. Here, we compare the overall proteomes from the carbon-starved (G-) and rapamycin treated (R+) model fungus Aspergillus nidulans. From about 1,100 visualized protein spots, we conservatively selected a total of 26 proteins with significant different expression. To highlight, increased levels of glucosidases and decreased levels of N-acetylglucosamine pyrophosphorylase were observed, suggesting degradation of the fungal cell wall as an alternate carbon source for both modes of induction. Cdc37 was reduced in expression while 14-3-3 ArtA was increased, implying regulation of polar growth, while also potentially regulating autophagy negatively via PKA or Tor. Other proteins included aspartate transaminase, tryptophan synthase B (TrpB), glycylpeptide N tetradecanoyltransferase (Nmt1), and aldehyde dehydrogenase (aldA). More interestingly, the majority of the identified proteins (16 of 26) were uniquely expressed in elevated levels in G-. A novel predicted protein from AN8223 which has no sequence homology to other organisms is also implicated to be involved in carbon-starvation. Thus, proteomic data here show that in A. nidulans, rapamycin induced autophagy and carbon-starvation induced autophagy share some effectors for cell survival, but predominantly involve different long-term effectors. PMID- 21618480 TI - Randomized phase II clinical trial of avotermin versus placebo for scar improvement. AB - BACKGROUND: Scarring is a major problem following skin injury. In early clinical trials, transforming growth factor beta3 (avotermin) improved scar appearance. The aim of this study was to determine whether an injection of avotermin at the time of wound closure is effective in improving scar appearance. METHODS: Study RN1001-0042, a double-blind, randomized, within-patient, placebo-controlled trial, investigated the efficacy and safety of four doses of avotermin given once. Patients undergoing bilateral surgery to remove varicose leg veins by saphenofemoral ligation and long saphenous vein stripping were enrolled at 20 European centres. A total of 156 patients were randomized to receive one of four doses of avotermin (5, 50, 200 or 500 ng per 100 ul, at 100 ul per linear cm of wound margin), administered by intradermal injection to the groin and distal wound margins of one leg; placebo was administered to the other leg. Scar appearance was evaluated by an independent panel of lay people (lay panel), investigators and patients. The primary efficacy variable was lay panel Total Scar Score (ToScar), derived from visual analogue scale scores for groin scars between 6 weeks and 7 months. RESULTS: Avotermin 500 ng significantly improved groin scar appearance compared with placebo (mean lay panel ToScar difference 16.49 mm; P = 0.036). CONCLUSION: Avotermin 500 ng per 100 ul per linear cm of wound margin given once is well tolerated and significantly improves scar appearance. PMID- 21618478 TI - Protein switches identified from diverse insertion libraries created using S1 nuclease digestion of supercoiled-form plasmid DNA. AB - We demonstrate that S1 nuclease converts supercoiled plasmid DNA to unit-length, linear dsDNA through the creation of a single, double-stranded break in a plasmid molecule. These double-stranded breaks occur not only in the origin of replication near inverted repeats but also at a wide variety of locations throughout the plasmid. S1 nuclease exhibits this activity under conditions typically employed for the nuclease's single-stranded nuclease activity. Thus, S1 nuclease digestion of plasmid DNA, unlike analogous digestion with DNaseI, effectively halts after the first double-stranded break. This property makes easier the construction of large domain insertion libraries in which the goal is to insert linear DNA at a variety of locations throughout a plasmid. We used this property to create a library in which a circularly permuted TEM1 beta-lactamase gene was inserted throughout a plasmid containing the gene encoding Escherichia coli ribose binding protein. Gene fusions that encode allosteric switch proteins in which ribose modulates beta-lactamase catalytic activity were isolated from this library using a combination of a genetic selection and a screen. PMID- 21618484 TI - Adverse effect of noise in the operating theatre on surgical-site infection. AB - BACKGROUND: The aim of this pilot study was to evaluate the noise level in an operating theatre as a possible surrogate marker for intraoperative behaviour, and to detect any correlation between sound level and subsequent surgical-site infection (SSI). METHODS: The sound level was measured during 35 elective open abdominal procedures. The noise intensity was registered digitally in decibels (dB) every second. A standard questionnaire was used to evaluate the behaviour of the surgical team during the operation. The primary outcome parameter was the SSI rate within 30 days of surgery. RESULTS: The overall rate of SSI was six of 35 (17 per cent). Demographic parameters and duration of operation were not significantly different between patients with, or without SSI. The median sound level (43.5 (range 26.0-60.0) versus 25.0 (25.0-60.0) dB; P = 0.040) and median level above baseline (10.7 (0.6-33.3) versus 0.6 (0.5-10.8); P = 0.001) were significantly higher for patients who developed a SSI. The sound level was at least 4 dB above the median in 22.5 per cent of the peaks in patients with SSI compared with 10.7 per cent in those without (P = 0.029). Talking about non surgery-related topics was associated with a significantly higher sound level (P = 0.024). CONCLUSION: Intraoperative noise volume was associated with SSI. This may be due to a lack of concentration, or a stressful environment, and may therefore represent a surrogate parameter by which to assess the behaviour of a surgical team. PMID- 21618488 TI - Volume analysis of outcome following restorative proctocolectomy (Br J Surg 2011; 98: 408-417). PMID- 21618489 TI - Nutritional predictors of postoperative outcome in pancreatic cancer (Br J Surg 2011; 98: 268-274). PMID- 21618491 TI - Abdominal trauma in primary blast injury (Br J Surg 2011; 98: 168-179). PMID- 21618494 TI - Effect of injury on S1 dorsal root ganglia in an experimental model of neuropathic faecal incontinence. AB - BACKGROUND: An experimental model of neuropathic faecal incontinence has recently been established. This study aimed to quantify and compare the effect of crush and compression injury on first-order sensory neurones of the inferior rectal nerve (IRN) using a nuclear marker of axonal injury, activating transcription factor (ATF) 3. METHODS: Eighteen Wistar rats were allocated to three groups: an unoperated control group, an IRN crush group (positive control) and a retrouterine balloon compression group. Five days after surgery, all animals were anaesthetized and perfused with fixative, and S1 dorsal root ganglia (DRG) were harvested. The tissue was sampled and neuronal nuclear ATF-3 expression calculated. RESULTS: Estimated total S1 DRG ATF-3 nuclear labelling was higher in the nerve crush (median (interquartile range) 171 (60-824) cells) and balloon compression (59 (20-274) cells) groups, compared with that in the unoperated control group (9 (3-24) cells) (P = 0.001 and P = 0.008 respectively). In all groups, most neurones displaying the marker of injury were of the C-fibre class. CONCLUSION: This study confirmed the presence of axonal injury in a pelvic compression model of obstetric injury. C-fibre afferent pathways appeared to be most vulnerable. Neuromodulation may function through augmentation of residual C fibre pathways. PMID- 21618495 TI - Recurrent groin hernia surgery. AB - BACKGROUND: The reoperation rate after recurrent groin hernia surgery is more than twice that recorded for primary groin hernia procedures. The aim was to define the outcome from routine redo hernia surgery by analysing a large population-based cohort from a national hernia register. METHODS: All recurrent groin hernia operations registered in the Swedish Hernia Register from 1992 to 2008 were analysed using multivariable analysis with stratification for preceding repair. RESULTS: Altogether 174,527 hernia operations were recorded in the Swedish Hernia Register between 1992 and 2008, including 19 582 reoperations. The preceding repair was included in the register for 5565 of these recurrent repairs. With laparoscopic repair as reference standard, the hazard ratio for recurrence was 2.55 (95 per cent confidence interval 1.66 to 3.93) after sutured repair, 1.53 (1.20 to 1.95) after Lichtenstein repair, 2.31 (1.76 to 3.03) after plug repair, 1.36 (0.95 to 1.94) after open preperitoneal mesh and 3.08 (2.22 to 4.29) after other repairs. Laparoscopic and open preperitoneal repair were associated with a lower risk of reoperation following a preceding open repair (P < 0.001), but no technique differed significantly from the others following a preceding preperitoneal repair. CONCLUSION: The laparoscopic and the open preperitoneal mesh methods of repair for recurrent groin hernias were associated with the lowest risk of reoperation. Although the method of repair in previous surgery must be considered, these techniques are the preferred methods for recurrent groin hernia surgery. PMID- 21618496 TI - Systematic review of outcome of downstaging hepatocellular cancer before liver transplantation in patients outside the Milan criteria. AB - BACKGROUND: The aim of this systematic review was to assess the evidence on tumour downstaging before liver transplantation in patients with hepatocellular carcinoma (HCC) initially staged beyond the Milan criteria. METHODS: MEDLINE (from 1952), Embase (from 1980) and the Cochrane Library were searched. The review included cohort studies that reported the outcomes of patients with HCC outside the Milan criteria who underwent downstaging before transplantation. RESULTS: Eight studies met the inclusion criteria and included a total of 720 patients who underwent transplantation following downstaging after initial presentation with disease outside the Milan criteria. The rate of successful downstaging varied from 24 to 69 per cent of patients. Reported survival rates ranged from 82 to 100 per cent, 79 to 100 per cent and 54.6 to 94 per cent at 1, 3 and 5 years respectively. These were comparable with results for patients presenting within the Milan criteria. CONCLUSION: Successful downstaging of HCC to within the Milan criteria is feasible in a proportion of patients. Absolute and disease-free survival rates in patients transplanted following downstaging are comparable to those in patients within the Milan criteria. PMID- 21618497 TI - Meta-analysis of two randomized controlled trials to identify long-term symptoms after division of the short gastric vessels during Nissen fundoplication. AB - BACKGROUND: Randomized trials suggest that division of the short gastric vessels during Nissen fundoplication is unnecessary. Some trials report an increased risk of gas bloat symptoms following division of the short gastric vessels. In this study long-term follow-up data from the two largest randomized clinical trials of division versus no division of the short gastric vessels during laparoscopic Nissen fundoplication were combined to determine whether there were differences in late outcome. METHODS: Patients with gastro-oesophageal reflux disease who underwent primary laparoscopic antireflux surgery and were included in two previously reported randomized trials were studied. Of 99 patients enrolled in the Swedish study and 102 in the Australian study, the short gastric vessels were divided in 104 and left intact in 97. Data sets were combined and late clinical outcomes analysed. RESULTS: At 10-12 years' follow-up (mean 11.5 years) clinical data were obtained from 170 patients (86 with vessels divided, 84 undivided). Statistical analysis of the combined data set showed no significant differences in symptoms of heartburn or dysphagia, ability to belch or vomit, and use of antisecretory medications. Division of the short gastric vessels was associated with a higher rate of bloating symptoms (72 versus 48 per cent; P = 0.002). CONCLUSION: Division of the short gastric vessels is followed by a slightly poorer clinical outcome at late follow-up after Nissen fundoplication. Surgeons should avoid dividing these vessels when undertaking a laparoscopic Nissen fundoplication. PMID- 21618498 TI - Cohort study of patients with adrenal lesions discovered incidentally. AB - BACKGROUND: This prospective cohort study investigated the incidence, clinical features and natural history of incidentally discovered adrenal mass lesions (adrenal incidentaloma, AI) in an unselected population undergoing radiological examination. METHODS: During an 18-month period, all patients with AI were reported prospectively from all 19 radiology departments in western Sweden. Inclusion criteria were: incidentally discovered adrenal enlargement or mass lesion in patients without extra-adrenal malignancy on detection. Clinical and biochemical evaluation was performed on inclusion and after 24 months. Computed tomography (CT) of the adrenals was scheduled at 4, 12 and 24 months. Magnetic resonance imaging was performed for lesions larger than 20 mm. The indications for surgical excision were: hormone activity, lesion diameter more than 30 mm, lesion growth or other radiological features suspicious of malignancy. RESULTS: Of 534 patients assessed for eligibility, 226 (mean age 67 years, 62.4 per cent women; mean lesion diameter 23.9 mm, 22.6 per cent bilateral) fulfilled the inclusion criteria. Mean follow-up was 19.0 months. After baseline evaluation, 14 patients had surgery owing to primary hyperaldosteronism (3), catecholamine producing tumour (1), tumour size (6), size and indication of subclinical hypercortisolism (3) and metastasis (1). No hypersecreting lesions were confirmed during follow-up; one patient underwent adrenalectomy for a suspected phaeochromocytoma (adrenocortical adenoma at histopathology). No primary adrenal malignancy was found. CONCLUSION: In this prospective cohort study 6.6 per cent of patients with an AI had surgery and benign hormone-producing tumours were verified in 3.1 per cent. Repeat CT and hormone evaluation after 2 years did not increase the sensitivity for diagnosis of malignant or hormone-producing tumours. PMID- 21618499 TI - Late follow-up of a randomized trial of routine duplex imaging before varicose vein surgery. AB - BACKGROUND: Routine preoperative duplex examination led to an improvement in results 2 years after surgery for primary varicose veins. The aim of the present study was to evaluate the impact of preoperative duplex imaging after 7 years, in relation to other risk factors for varicose vein recurrence. METHODS: Patients with primary varicose veins were randomized to operation with (group 1), or without (group 2) preoperative duplex imaging. The same patients were invited to attend follow-up with interview, clinical examination and duplex imaging. Quality of life (QoL) was measured with the Short Form 36 questionnaire. RESULTS: Some 293 patients (343 legs) were included initially; after 7 years 227 were interviewed, or their records reviewed: 114 in group 1 and 113 in group 2. One hundred and ninety-four legs (95 in group 1 and 99 in group 2) were examined clinically and with duplex imaging. Incompetence was seen at the saphenofemoral junction and/or saphenopopliteal junction in 14 per cent of legs in group 1 and 46 per cent in group 2 (P < 0.001). QoL was similar in both groups. After a mean follow-up of 7 years (and including patients who underwent surgery after the review), 15 legs in group 1 needed reoperation and 38 in group 2 (P = 0.001). CONCLUSION: Routine preoperative duplex imaging improved the results of surgery for primary varicose veins for at least 7 years. PMID- 21618503 TI - Cancer immunotherapy: Progress and challenges in the clinical setting. PMID- 21618505 TI - Transduced B cells: B is for 'beneficial'! AB - B cells have been used as tolerogenic APCs for nearly two decades. However, the ability to transduce B cells for use in gene therapy has been hampered by the low efficiency of transduction of resting B cells. This has been partially overcome by mitogenic activation of these cells, a factor that is not without risks as activated B cells may become pathogenic. In this issue of the European Journal of Immunology, this challenge is met by achieving high-efficiency transduction of resting murine B cells with a lentiviral vector. Furthermore, the application of this protocol to generate MOG-expressing B cells and successfully prevent EAE, as described in this issue, is an important step forward in B-cell therapy. PMID- 21618506 TI - IL-33 links tissue cells, dendritic cells and Th2 cell development in a mouse model of asthma. AB - IL-33 is becoming a central molecule in allergic asthma that addresses various cascades of innate and adaptive immune responses that lead to inflammation in the lung. Its effects are exerted via its heterodimeric receptor that consists of ST2 and the ubiquitously expressed IL-1 receptor accessory protein (ILRAcP). IL-33 integrates both innate and adaptive immunity in a unique fashion via basophils, mast cells, eosinophils, innate lymphoid cells, NK and NKT cells, nuocytes, Th2 lymphocytes and a CD34(pos) precursor cell population. These actions of IL-33 seem to be particularly strong and dominant in models with mucosal inflammation. A study in this issue of the European Journal of Immunology demonstrates that IL 33 acts, in an ST2-dependent manner, as a maturation factor for BM-derived DCs via up-regulation of CD80, CD40 and OX40L. This process is accompanied by the release of pro-inflammatory cytokines, such as IL-6, IL-1beta, TNF-alpha and TARC/CCL17. IL-33-pre-treated DCs were significantly more potent for the generation of allergen-specific Th2-type cells with IL-5 and IL-13 production. Intratracheal administration of OVA-pulsed DCs with IL-33 significantly enhances eosinophil numbers and mucous secretion. In conclusion, IL-33 affects both the development of allergic sensitization and the development of lung inflammation in allergic asthma. PMID- 21618507 TI - Dual inhibition of phosphatidylinositol 3-kinase/Akt and mammalian target of rapamycin signaling in human nonsmall cell lung cancer cells by a dietary flavonoid fisetin. AB - Lung cancer is one of the most commonly occurring malignancies. It has been reported that mammalian target of rapamycin (mTOR) is phosphorylated in lung cancer and its activation was more frequent in tumors with overexpression of phosphatidylinositol 3-kinase (PI3K)/Akt. Therefore, dual inhibitors of PI3K/Akt and mTOR signaling could be valuable agents for treating lung cancer. In the present study, we show that fisetin, a dietary tetrahydroxyflavone inhibits cell growth with the concomitant suppression of PI3K/Akt and mTOR signaling in human nonsmall cell lung cancer (NSCLC) cells. Using autodock 4, we found that fisetin physically interacts with the mTOR complex at two sites. Fisetin treatment was also found to reduce the formation of A549 cell colonies in a dose-dependent manner. Treatment of cells with fisetin caused decrease in the protein expression of PI3K (p85 and p110), inhibition of phosphorylation of Akt, mTOR, p70S6K1, eIF 4E and 4E-BP1. Fisetin-treated cells also exhibited dose-dependent inhibition of the constituents of mTOR signaling complex such as Rictor, Raptor, GbetaL and PRAS40. There was an increase in the phosphorylation of AMPKalpha and a decrease in the phosphorylation of TSC2 on treatment of cells with fisetin. We also found that treatment of cells with mTOR inhibitor rapamycin and mTOR-siRNA caused decrease in phosphorylation of mTOR and its target proteins which were further downregulated on treatment with fisetin, suggesting that these effects are mediated in part, through mTOR signaling. Our results show that fisetin suppressed PI3K/Akt and mTOR signaling in NSCLC cells and thus, could be developed as a chemotherapeutic agent against human lung cancer. PMID- 21618508 TI - Inhibition of VEGF induces cellular senescence in colorectal cancer cells. AB - Vascular endothelial growth factor (VEGF) inhibitors, such as bevacizumab, have improved outcomes in metastatic colorectal cancer (CRC). Recent studies have suggested that VEGF can delay the onset of cellular senescence in human endothelial cells. As VEGF receptors are known to be upregulated in CRC, we hypothesized that VEGF inhibition may directly influence cellular senescence in this disease. In our study, we observed that treatment with bevacizumab caused a significant increase (p < 0.05) in cellular senescence in vitro in several CRC cells, such as MIP101, RKO, SW620 and SW480 cells, compared to untreated or human IgG-treated control cells. Similar results were also obtained from cells treated with a VEGFR2 kinase inhibitor Ki8751. In vivo, cellular senescence was detected in MIP101 tumor xenografts from 75% of mice treated with bevacizumab, while cellular senescence was undetectable in xenografts from mice treated with saline or human IgG (p < 0.05). Interestingly, we also observed that the proportion of senescent cells in colon cancer tissues obtained from patients treated with bevacizumab was 4.4-fold higher (p < 0.01) than those of untreated patients. To understand how VEGF inhibitors may regulate cellular senescence, we noted that among the two important regulators of senescent growth arrest of tumor cells, bevacizumab-associated increase in cellular senescence coincided with an upregulation of p16 but appeared to be independent of p53. siRNA silencing of p16 gene in MIP101 cells suppressed bevacizumab-induced cellular senescence, while silencing of p53 had no effect. These findings demonstrate a novel antitumor activity of VEGF inhibitors in CRC, involving p16. PMID- 21618509 TI - Pathological complete remission in patients with oesophagogastric cancer receiving preoperative 5-fluorouracil, oxaliplatin and docetaxel. AB - The aim of this study was to determine the pathological complete remission (pCR) rate, and its relationship to clinical outcome, in patients with adenocarcinoma of the stomach or oesophagogastric junction receiving preoperative 5 fluorouracil, leucovorin, oxaliplatin and docetaxel (FLOT) every 2 weeks. Data from these patients who received at least one cycle of preoperative FLOT followed by surgery were prospectively collected in three German centres. Outcome analyses were conducted and tumour samples were evaluated for pathological remission by a central pathologist. A total of 46 patients were included in this analysis. All patients had clinical T3- and/or N+-stages and 11 (23.9%) had distant metastases (M1). After a median of 4 (range 2-8) preoperative cycles, 8 of 46 patients (17.4%) achieved a pCR. The pCR rate was highest in tumours of intestinal type histology (30.8%) and in those located in the oesophagogastric junction (30.4%) and lowest in patients with diffuse/mixed type tumours (0%) or tumours located in the stomach (4.3%; p < 0.05 for both comparisons). Patients with pCR had 100% probability of overall and disease-free survival (DFS) during the observation period, which was significantly higher (p = 0.037 and p = 0.009, respectively) than the survival probability in patients without pCR. In conclusion, treatment intensification using FLOT was associated with significant pCR rates in patients with oesophagogastric cancer. The distribution of pCR appeared to be significantly different according to histological type and location of the tumours. PMID- 21618510 TI - Cetuximab +/- chemotherapy enhances dendritic cell-mediated phagocytosis of colon cancer cells and ignites a highly efficient colon cancer antigen-specific cytotoxic T-cell response in vitro. AB - Cetuximab is a human/mouse chimeric IgG1 monoclonal antibody (mAb) to epidermal growth factor receptor, approved for colorectal carcinoma treatment in combination with chemotherapy. The immune-mediated effects elicited by its human fraction of crystallization moiety might critically contribute to the overall anti-tumor effectiveness of the antibody. We therefore investigated cetuximab ability to promote colon cancer cell opsonization and phagocytosis by human dendritic cells (DCs) that are subsequently engaged in antigen-cross presentation to cytotoxic T-lymphocyte (CTL) precursors. Human colon cancer cell lines were evaluated for susceptibility to DC-mediated phagocytosis before and after treatment with chemotherapy +/- cetuximab in vitro. Human DCs loaded with control or drug-treated cetuximab-coated colon cancer cells were used to in vitro generate cytotoxic T cell clones from peripheral blood mononuclear cells of human leucocyte antigen-A(*)02.01(+) donors. T-cell cultures were characterized for immune-phenotype and tumor-antigen specific CTL activity. The results confirmed that treatment of tumor cells with irinotecan + L-folinate + 5-flurouracil (ILF) or with gemcitabine + ILF increased tumor antigen expression. Moreover, malignant cells exposed to chemotherapy and cetuximab were highly susceptible to phagocytosis by human DCs and were able to promote their activation. The consequent DC-mediated cross-priming of antigens derived from mAb-covered/drug treated cancer cells elicited a robust CTL anti-tumor response. On the basis of our data, we suggest a possible involvement of CTL-dependent immunity in cetuximab anti-cancer effects. PMID- 21618511 TI - Lymph node CEA and MUC2 mRNA as useful predictors of outcome in colorectal cancer. AB - The aim was to explore the utility for staging and prognostic impact of carcinoembryonic antigen (CEA), cytokeratin 20 (CK20), guanylyl cyclase C (GCC), CUB (complement protein subcomponents C1r/C1s, urchin embryonic growth factor, and bone morphogenic protein 1) containing domain protein 1 (CDCP1) and mucin 2 (MUC2) mRNA levels in mesenteric lymph nodes of colorectal cancer (CRC) patients. Lymph nodes were collected at surgery and bisected; one half was subjected to biomarker mRNA analysis using real-time quantitative RT-PCR and the other half to routine histopathology. Lymph nodes from 174 CRC patients and 24 controls were analyzed. The median follow-up time was 59 (range 17-131) months. Cut-off levels were defined by analyzing quintiles by Cox regression model. CEA mRNA showed the best discriminating power between patients with recurrence in CRC after surgery and patients who were apparently disease-free (p = 0.015). The risk of recurrence for the CEA(+) patients was 4.6 times greater than for the CEA(-) patients (p < 0.0001). The other biomarkers gave lower hazard ratios. Cumulative survival analysis demonstrated that the average survival time was 99 months for CEA(-) patients compared to 39 months for CEA(+) patients, a difference of 60 months (p < 0.0001). Six to nine percent of the Stage I and Stage II patients [H&E(-)] had CEA(+), CK20(+), GCC(+) and/or MUC2(+) lymph nodes. Two of these patients died from recurrent CRC. Low lymph node MUC2/CEA mRNA ratio identified patients with high risk for recurrence (p = 0.011). Thus, quantitative reverse transcriptase polymerase chain reaction of CEA mRNA is a sensitive method to identify tumor cells in lymph nodes of CRC patients and, in combination with MUC2 mRNA, allows improved prediction of clinical outcome. PMID- 21618512 TI - Cisplatin increases B-cell-lymphoma-2 expression via activation of protein kinase C and Akt2 in endometrial cancer cells. AB - Human carcinomas often show resistance to cisplatin and Bcl-2 is associated with resistance to cisplatin. However, Bcl-2 regulation on cisplatin treatment in human cancers is unknown. Here, we show a novel mechanism by which cisplatin treatment promotes resistance by increasing the expression of Bcl-2 mRNA. Bcl-2 mRNA and protein expression was increased in cisplatin-resistant endometrial cancer cell lines (KLE and HEC-1-A), but not in cisplatin-sensitive cell line (Ishikawa). Cisplatin-mediated increase in Bcl-2 expression was blocked by combination with either actinomycin D or cycloheximide. In addition, Bcl-2 inhibition by HA14-1 led to increased cisplatin-induced apoptosis in KLE and HEC 1-A, whereas Bcl-2 overexpression in Ishikawa led to decreased cisplatin-induced apoptosis. Inhibition of protein kinase C (PKC) activity prevented cisplatin dependant increase in Bcl-2 mRNA, and induced apoptosis in KLE cells. Furthermore, PKC inhibition was associated with decreased Akt and NF-kappaB activity. Cells stably expressing shRNA for Akt isoforms revealed that Akt2 was involved in cisplatin-dependant increase in Bcl-2 and apoptosis. Overexpression of Akt2 in Akt2-deficient cells led to increased Bcl-2 expression on cisplatin treatment. Our data suggest a novel regulation pathway of Bcl-2 by cisplatin, via the activation of PKC and Akt2, which has a profound impact on resistance to cisplatin-induced apoptosis in endometrial cancer cells. PMID- 21618513 TI - Dietary lignan and proanthocyanidin consumption and colorectal adenoma recurrence in the Polyp Prevention Trial. AB - Lignans and proanthocyanidins are plant polyphenols that have shown protective properties against colorectal neoplasms in some human studies. Using logistic regression, we estimated odds ratios (ORs) and 95% confidence intervals (CIs) to prospectively evaluate the association between lignan and proanthocyanidin intake, estimated from databases linked to a food frequency questionnaire, and adenoma recurrence in 1,859 participants of the Polyp Prevention Trial. Overall, individual or total lignans or proanthocyanidins were not associated with colorectal adenoma recurrence. However, in sex-specific analyses, total lignan intake was positively associated with any adenoma recurrence in women (highest vs. lowest lignan intake quartile OR = 2.07, 95% CI: 1.22-3.52, p trend = 0.004) but not in men (p interaction = 0.04). To conclude, dietary lignan and proanthocyanidin consumption were not generally related to colorectal adenoma recurrence; however, high lignan intake may increase the risk of adenoma recurrence in women. PMID- 21618514 TI - High-risk human papillomavirus in oral squamous cell carcinoma of young patients. AB - The aim of this study was to investigate a possible relation between oral squamous cell carcinoma (SCC), the presence of high-risk human papillomavirus (HR HPV) DNA and p16 expression in young patients. Paraffin-embedded tumor blocks from 47 oral SCC of young (<=40-year old) patients were evaluated. The presence of HPV DNA in tumor specimens was analyzed by polymerase chain reaction (PCR) using GP5+/GP6+ generic primers (L1 region) followed by dot blot hybridization for HPV typing. When necessary, the HPV16 positivity was confirmed by PCR HPV16 E7-specific primers. Cases involving young patients were compared with 67 oral SCC from patients >=50-year old (controls). Demographic and clinical data were collected to analyze patient outcomes. p16(ink4) expression was evaluated by immunostaining of tissue microarrays. HPV16 was detected in 22 (19.2%) cases; 15 (68.2%) young and 7 (31.8%) control patients, a statistically significant difference (p = 0.01). In 1 (1.7%) young group specimen, HPV DNA 16 and 18 was detected. p16 expression was observed in 11 (25.6%) cases from the young group and in 11 (19.6%) controls (p = 0.48). Association between HPV and p16 was verified, and it was statistically significant (p = 0.002). The higher prevalence of high-risk HPV types, especially HPV16, may be a contributing factor to oral carcinogenesis in younger individuals. PMID- 21618515 TI - Is mortality after childhood cancer dependent on social or economic resources of parents? A population-based study. AB - Diagnostic and treatment protocols for childhood cancer are generally standardized, and therefore, survival ought to be fairly equal across social strata in societies with free public health care readily available. Nevertheless, our study explores whether there are disparities in mortality after childhood cancer in Norway depending on socioeconomic status of parents. Limited knowledge on differentials exists from earlier analyses. Discrete-time hazard regression models for all-cause mortality for the first 10 years after diagnosis were estimated for all Norwegian children (younger than 20 years), who were diagnosed with cancer during 1974-2007 (N = 6,280), using data from five national registers. Mortality was reduced by about 15% for children with highly educated mothers and children without siblings. These effects were most pronounced for cancers predicted to encompass intense, long-lasting treatments resulting in chronic health problems. Neither earnings nor the marital status of parents affected children's survival. This large, registry-based study suggests that time constraints and various noneconomic rewards of parents from their education appears to have an impact on childhood cancer survival. It may be that children with resourceful parents are healthier at the outset and/or are more likely to avoid later health problems. It may also be that children of well-informed and strongly involved parents are offered better treatment or are able to make better use of what is offered, for instance, by adhering more closely to recommendations for follow-up treatment. The possibility of such differentials in offered and actual treatment should be addressed in future research. PMID- 21618516 TI - MUC4: a novel prognostic factor of oral squamous cell carcinoma. AB - MUC4 mucin is now known to be expressed in various normal and cancer tissues. We have previously reported that MUC4 expression is a novel prognostic factor in several malignant tumors; however, it has not been investigated in oral squamous cell carcinoma (OSCC). The aim of our study is to evaluate the prognostic significance of MUC4 expression in OSCC. We examined the expression profile of MUC4 in OSCC tissues from 150 patients using immunohistochemistry. Its prognostic significance in OSCC was statistically analyzed. MUC4 was expressed in 61 of the 150 patients with OSCC. MUC4 expression was significantly correlated with higher T classification (p = 0.0004), positive nodal metastasis (p = 0.049), advanced tumor stage (p = 0.002), diffuse invasion of cancer cells (p = 0.004) and patient's death (p = 0.004) in OSCC. Multivariate analysis showed that MUC4 expression (p = 0.011), tumor location (p = 0.032) and diffuse invasion (p = 0.009) were statistically significant risk factors. Backward stepwise multivariate analysis demonstrated MUC4 expression (p = 0.0015) and diffuse invasion (p = 0.018) to be statistically significant independent risk factors of poor survival in OSCC. The disease-free and overall survival of patients with MUC4 expression was significantly worse than those without MUC4 expression (p < 0.0001 and p = 0.0001). In addition, the MUC4 expression was a significant risk factor for local recurrence and subsequent nodal metastasis in OSCC (p = 0.017 and p = 0.0001). We first report MUC4 overexpression is an independent factor for poor prognosis of patients with OSCC; therefore, patients with OSCC showing positive MUC4 expression should be followed up carefully. PMID- 21618517 TI - Expression and pharmacological inhibition of thymidylate synthase and Src kinase in nonsmall cell lung cancer. AB - The combination of cytotoxic chemotherapy with signaling pathway inhibitors represents a potential strategy to improve the treatment of nonsmall cell lung cancer (NSCLC). Thymidylate synthase (TS) is an enzyme essential for DNA synthesis, and its overexpression has been associated with the reduced sensitivity to antifolate agents. Src is a tyrosine kinase that modulates the cytotoxicity of cancer cells after drug treatment, and in vitro data indicate that its inhibition could revert the resistance to TS-inhibiting drugs. Our study investigated the significance of TS and Src expression in NSCLC tissues, and the effects of their pharmacological inhibition in cell lines. In tumor and normal tissues from 94 resected NSCLC patients, TS and Src transcript levels were found positively correlated (R(S) = 0.66), associated with patients smoking history and overall survival. At multivariate analysis, TS gene expression was an independent prognostic factor (relative risk (RR) = 1.78, from 1.16 to 2.72; p < 0.01). Immunohistochemical detection in tumor specimens confirmed that Src kinase activation, evaluated by phospho-specific antibody, was associated to a higher TS expression. In cell lines, dasatinib, a Src-inhibiting agent, synergistically enhanced pemetrexed-cytotoxicity of A549 cells, as evaluated by MTT and apoptosis assays. The biological explanation for this interaction was based on the upregulation of TS messenger RNA and protein levels induced by pemetrexed, which was significantly prevented by dasatinib cotreatment. The data of our study suggest that TS and Src may belong to a common pathway that bears prognostic significance in NSCLC, and that Src represents a potential target to improve the efficacy of TS-inhibiting agents. PMID- 21618518 TI - Kinesin family member 14: an independent prognostic marker and potential therapeutic target for ovarian cancer. AB - The novel oncogene KIF14 (kinesin family member 14) shows genomic gain and overexpression in many cancers including OvCa (ovarian cancer). We discovered that expression of the mitotic kinesin KIF14 is predictive of poor outcome in breast and lung cancers. We now determine the prognostic significance of KIF14 expression in primary OvCa tumors, and evaluate KIF14 action on OvCa cell tumorigenicity in vitro. Gene-specific multiplex PCR and real-time QPCR were used to measure KIF14 genomic (109 samples) and mRNA levels (122 samples) in OvCa tumors. Association of KIF14 with clinical variables was studied using Kaplan Meier survival and Cox regression analyses. Cellular effects of KIF14 overexpression (stable transfection) and inhibition (stable shRNA knockdown) were studied by proliferation (cell counts), survival (Annexin V immunocytochemistry) and colony formation (soft-agar growth). KIF14 genomic gain (>2.6 copies) was present in 30% of serous OvCas, and KIF14 mRNA was elevated in 91% of tumors versus normal epithelium. High KIF14 in tumors independently predicted for worse outcome (p = 0.03) with loss of correlation with proliferation marker expression and increased rates of recurrence. Overexpression of KIF14 in OvCa cell lines increased proliferation and colony formation (p < 0.01), whereas KIF14 knockdown induced apoptosis and dramatically reduced colony formation (p < 0.05). Our findings indicate that KIF14 mRNA is an independent prognostic marker in serous OvCa. Dependence of OvCa cells on KIF14 for maintenance of in vitro colony formation suggests a role of KIF14 in promoting a tumorigenic phenotype, beyond its known role in proliferation. PMID- 21618519 TI - Let-7 modulates acquired resistance of ovarian cancer to Taxanes via IMP-1 mediated stabilization of multidrug resistance 1. AB - Ovarian cancer patients frequently develop resistance to chemotherapy regiments using Taxol and carboplatin. One of the resistance factors that protects cancer cells from Taxol-based therapy is multidrug resistance 1 (MDR1). micro(mi)RNAs are small noncoding RNAs that negatively regulate protein expression. Members of the let-7 family of miRNAs are downregulated in many human cancers, and low let-7 expression has been correlated with resistance to microtubule targeting drugs (Taxanes), although little is known that would explain this activity. We now provide evidence that, although let-7 is not a universal sensitizer of cancer cells to Taxanes, it affects acquired resistance of cells to this class of drugs by targeting IMP-1, resulting in destabilization of the mRNA of MDR1. Introducing let-7g into ADR-RES cells expressing both IMP-1 and MDR1 reduced expression of both proteins rendering the cells more sensitive to treatment with either Taxol or vinblastine without affecting the sensitivity of the cells to carboplatin, a non-MDR1 substrate. This effect could be reversed by reintroducing IMP-1 into let 7g high/MDR1 low cells causing MDR1 to again become stabilized. Consistently, many relapsed ovarian cancer patients tested before and after chemotherapy were found to downregulate let-7 and to co-upregulate IMP-1 and MDR1, and the increase in the expression levels of both proteins after chemotherapy negatively correlated with disease-free time before recurrence. Our data point at IMP-1 and MDR1 as indicators for response to therapy, and at IMP-1 as a novel therapeutic target for overcoming multidrug resistance of ovarian cancer. PMID- 21618520 TI - Male circumcision is associated with a lower prevalence of human papillomavirus associated penile lesions among Kenyan men. AB - Human papillomavirus (HPV)-associated penile lesions in men may increase the risk of HPV transmission to their female partners. Risk factor data on HPV-associated penile lesions are needed from regions with a high burden of cervical cancer. Visual inspection of the penis was conducted using a colposcope at the 24-month visit among participants in a randomized controlled trial of male circumcision in Kenya, from May 2006 to October 2007. All photos were read independently by two observers for quality control. Penile exfoliated cells sampled from the glans/coronal sulcus and the shaft were tested for HPV DNA using GP5+/6+ PCR and for HPV16, 18 and 31 viral loads using a real time PCR assay. Of 275 men, 151 were circumcised and 124 uncircumcised. The median age was 22 years. Circumcised men had a lower prevalence of flat penile lesions (0.7%) versus uncircumcised (26.0%); adjusted odds ratio (OR) = 0.02; 95% confidence interval (CI) = 0.003 0.1. Compared to men who were HPV negative, men who were HPV DNA positive (OR = 6.5; 95% CI = 2.4-17.5) or who had high HPV16/18/31 viral load (OR = 5.2; 95% CI = 1.1-24.4) had higher odds of flat penile lesions. Among men with flat penile lesions, HPV56 (29.0%) and 16 (25.8%) were the most common types within single or multiple infections. Flat penile lesions are much more frequent in uncircumcised men and associated with higher prevalence of HPV and higher viral loads. This study suggests that circumcision reduces the prevalence of HPV-associated flat lesions and may ultimately reduce male-to-female HPV transmission. PMID- 21618521 TI - Demethylation of the FOXP3 gene in human melanoma cells precludes the use of this epigenetic mark for quantification of Tregs in unseparated melanoma samples. AB - The human suppressive T cells that stably express transcription factor FOXP3, or regulatory T cells (Tregs), are thought to suppress antitumor immune responses. The most specific marker for human Tregs is the demethylation of CpG dinucleotides located in the first intron of FOXP3 (FOXP3i1). FOXP3i1 is completely methylated in other hematopoietic cells, including nonsuppressive T cells that transiently express FOXP3 after activation. Previously, we and others reported estimations of the frequency of Tregs in the blood of melanoma patients using a FOXP3i1 methylation-specific qPCR assay. Here, we attempted to quantify Tregs inside tumor samples using this assay. However, we found demethylated FOXP3i1 sequences in the melanoma cells themselves. This demethylation was not associated with substantial FOXP3 mRNA or protein expression, even though the demethylation extended to the promoter and terminal regions of the gene in some melanoma cells. Our results imply that analyzing Treg frequencies by quantification of demethylated FOXP3i1 will require that tumor-infiltrating T cells be separated from melanoma cells. PMID- 21618522 TI - Carcinogen metabolism genes, red meat and poultry intake, and colorectal cancer risk. AB - Diets high in red meat are established risk factors for colorectal cancer (CRC). Carcinogenic compounds generated during meat cooking have been implicated as causal agents. We conducted a family-based case-control study to investigate the association between polymorphisms in carcinogen metabolism genes (CYP1A2 -154A>C, CYP1B1 Leu432Val, CYP2E1 -1054C>T, GSTP1 Ile105Val, PTGS2 5UTR -765, EPHX1 Tyr113His, NAT2 Ile114Thr, NAT2 Arg197Gln and NAT2 Gly286Glu) and CRC risk. We tested for gene-environment interactions using case-only analyses (N = 577) and compared statistically significant results to those obtained using case unaffected sibling comparisons (N = 307 sibships). Our results suggested that CYP1A2 -154A>C might modify the association between intake of red meat cooked using high temperature methods and well done on the inside and CRC risk (case only interaction OR = 1.53; 95% CI = 1.19-1.97; p = 0.0008) and the association between intake of red meat heavily browned on the outside and rectal cancer risk (case-only interaction OR = 0.65; 95% CI = 0.48-0.86; p = 0.003). We also found that GSTP1 Ile105Val might modify the association between intake of poultry cooked with high temperature methods and CRC risk (p = 0.0035), a finding that was stronger among rectal cancer cases. Our results support a role for heterocyclic amines that form in red meat as a potential explanation for the observed association between diets high in red meat and CRC. Our findings also suggest a possible role for diets high in poultry cooked at high temperatures in CRC risk. PMID- 21618523 TI - MAGE-D4B is a novel marker of poor prognosis and potential therapeutic target involved in breast cancer tumorigenesis. AB - Melanoma-associated antigen (MAGE) family members are generally described as tumor-specific antigens. An association between MAGE-D4B and breast cancer has yet to be reported and the functional role of the encoded protein has never been established. We performed microarray analysis of 104 invasive breast tumors and matched non-cancerous breast biopsies. qPCR was used for validation in an independent biobank. To investigate the biological relevance of MAGE-D4B in breast tumorigenesis, its phenotypic effects were assessed in vitro. Overall, MAGE-D4B was detected in 43% of tumors while undetected in normal breast tissue. MAGE-D4B was found to correlate with tumor progression and to be an independent prognostic marker for poor outcome in term of relapse-free and overall survival, with potential predictive relevance in relation to response to chemotherapy. RNA interference-mediated knockdown of MAGE-D4B significantly hampered the invasive properties of Hs578T cells by affecting anchorage-independent growth, adhesion, migration and invasion affecting anchorage-independent growth, adhesion, migration and invasion and by modulating expression of invasion-suppressor gene E cadherin. PMID- 21618524 TI - IGF binding protein-6 expression in vascular endothelial cells is induced by hypoxia and plays a negative role in tumor angiogenesis. AB - Hypoxia stimulates tumor angiogenesis by inducing the expression of angiogenic molecules. The negative regulators of this process, however, are not well understood. Here, we report that hypoxia induced the expression of insulin-like growth factor binding protein-6 (IGFBP-6), a tumor repressor, in human and rodent vascular endothelial cells (VECs) via a hypoxia-inducible factor (HIF)-mediated mechanism. Addition of human IGFBP-6 to cultured human VECs inhibited angiogenesis in vitro. An IGFBP-6 mutant with at least 10,000-fold lower binding affinity for IGFs was an equally potent inhibitor of angiogenesis, suggesting that this action of IGFBP-6 is IGF-independent. The functional relationship between IGFBP-6 and vascular endothelial growth factor (VEGF), a major hypoxia inducible angiogenic molecule, was examined. While VEGF alone increased angiogenesis in vitro, co-incubation with IGFBP-6 abolished VEGF-stimulated angiogenesis. The in vivo role of IGFBP-6 in angiogenesis was tested in flk1:GFP zebrafish embryos, which exhibit green fluorescence protein in developing vascular endothelium, permitting visualization of developing blood vessels. Injection of human IGFBP-6 mRNA reduced the number of embryonic inter-segmental blood vessels by ~40%. This anti-angiogenic activity is conserved in zebrafish because expression of zebrafish IGFBP-6b had similar effects. To determine the anti-angiogenic effect of IGFBP-6 in a tumor model, human Rh30 rhabdomyosarcoma cells stably transfected with IGFBP-6 were inoculated into athymic BALB/c nude mice. Vessel density was 52% lower in IGFBP-6-transfected xenografts than in vector control xenografts. These results suggest that the expression of IGFBP-6 in VECs is up-regulated by hypoxia and IGFBP-6 inhibits angiogenesis in vitro and in vivo. PMID- 21618525 TI - Induction of torpor: mimicking natural metabolic suppression for biomedical applications. AB - Mammalian hibernation consists of periods of depressed metabolism and reduced body temperature called "torpor" that are interspersed by normothermic arousal periods. Numerous cellular processes are halted during torpor, including transcription, translation, and ion homeostasis. Hibernators are able to survive long periods of low blood flow and body temperature followed by rewarming and reperfusion without overt signs of organ injury, which makes these animals excellent models for application of natural protective mechanisms to human medicine. This review examines efforts to induce torpor-like states in non hibernating species using pharmacological compounds. Elucidating the underlying mechanisms of natural and pharmacologically induced torpor will speed the development of new clinical approaches to treat a variety of trauma and stress states in humans. PMID- 21618526 TI - Hepatocyte growth factor preferentially activates the anti-inflammatory arm of NF kappaB signaling to induce A20 and protect renal proximal tubular epithelial cells from inflammation. AB - Inflammation induces the NF-kappaB dependent protein A20 in human renal proximal tubular epithelial cells (RPTEC), which secondarily contains inflammation by shutting down NF-kappaB activation. We surmised that inducing A20 without engaging the pro-inflammatory arm of NF-kappaB could improve outcomes in kidney disease. We showed that hepatocyte growth factor (HGF) increases A20 mRNA and protein levels in RPTEC without causing inflammation. Upregulation of A20 by HGF was NF-kappaB/RelA dependent as it was abolished by overexpressing IkappaBalpha or silencing p65/RelA. Unlike TNFalpha, HGF caused minimal IkappaBalpha and p65/RelA phosphorylation, with moderate IkappaBalpha degradation. Upstream, HGF led to robust and sustained AKT activation, which was required for p65 phosphorylation and A20 upregulation. While HGF treatment of RPTEC significantly increased A20 mRNA, it failed to induce NF-kappaB dependent, pro-inflammatory MCP 1, VCAM-1, and ICAM-1 mRNA. This indicates that HGF preferentially upregulates protective (A20) over pro-inflammatory NF-kappaB dependent genes. Upregulation of A20 supported the anti-inflammatory effects of HGF in RPTEC. HGF pretreatment significantly attenuated TNFalpha-mediated increase of ICAM-1, a finding partially reversed by silencing A20. In conclusion, this is the first demonstration that HGF activates an AKT-p65/RelA pathway to preferentially induce A20 but not inflammatory molecules. This could be highly desirable in acute and chronic renal injury where A20-based anti-inflammatory therapies are beneficial. PMID- 21618527 TI - Attenuation of microRNA-22 derepressed PTEN to effectively protect rat cardiomyocytes from hypertrophy. AB - Cardiac hypertrophy, which is characterized by the enlargement of cell size, reactivation of fetal genes, remains one of the most important triggers to heart failure. Increasing evidence shows that microRNA (miRNA) is extensively involved in the pathogenesis of cardiac hypertrophy. But the effects of miRNAs on cardiomyocyte hypertrophy have not been completely solved yet. Here, we showed that a collection of miRNAs was aberrantly expressed in hypertrophic cardiomyocytes induced by phenylephrine (PE) or angiotensin II (Ang II). Among them, miR-22 was the most strikingly up-regulated miRNA. To investigate the role of miR-22 in hypertrophy, both over-expression and knock-down assays were performed on cardiomyocytes. The results showed that up-regulation of miR-22 significantly increased the cell size and markedly influenced the expression of hypertrophic markers, including induction of nppa and reduction of myh6. In contrast, reduction of miR-22 level attenuated either PE- or Ang II-induced hypertrophic reaction. Furthermore, several genes, including PTEN, were identified as potential targets of miR-22 by bioinformatic algorithms. Using luciferase analysis, miR-22 could significantly suppress the luciferase activity of reporter fused with 3' untranslated region of PTEN mRNA. Furthermore, up regulation of miR-22 could suppress the protein level of PTEN and reduction of miR-22 level markedly increased the protein level of PTEN in cardiomyocytes by Western blot analysis, suggesting that the contribution of miR-22 to cardiomyocyte hypertrophy may be partially through targeting PTEN. Taken together, miRNAs were dynamically regulated in cardiomyocyte hypertrophy and attenuation of miR-22 in rat cardiomyocytes efficiently protected from hypertrophic effects through derepressing PTEN. PMID- 21618528 TI - Cdc42 negatively regulates intrinsic migration of highly aggressive breast cancer cells. AB - The small GTPase Cdc42 has been implicated as an important regulator of cell migration. However, whether Cdc42 plays similar role in all cancer cells irrespective of metastatic potential remains poorly defined. Here, we show by using three different breast cancer cell lines with different metastatic potential, the role of Cdc42 in cell migration/invasion and its relationship with a number of downstream signaling pathways controlling cell migration. Small interfering RNA (siRNA)-mediated knockdown of Cdc42 in two highly metastatic breast cancer cell lines (MDA-MB-231 and C3L5) resulted in enhancement, whereas the same in moderately metastatic (Hs578T) cell line resulted in inhibition of intrinsic cellular migration/invasion. Furthermore, Cdc42 silencing in MDA-MB-231 and C3L5 but not Hs578T cells was shown to be accompanied by increased RhoA activity and phosphorylation of protein kinase C (PKC)-delta, extracellular signal regulated kinase1/2 (Erk1/2), and protein kinase A (PKA). Pharmacological inhibition of PKCdelta, MEK-Erk1/2, or PKA was shown to inhibit migration of both control and Cdc42-silenced MDA-MB-231 cells. Furthermore, introduction of constitutively active Cdc42 was shown to decrease migration/invasion of MDA-MB 231 and C3L5 but increase migration/invasion of Hs578T cells. This decreased migration/invasion of MDA-MB-231 and C3L5 cells was also shown to be accompanied by the decrease in the phosphorylations of PKCdelta, Erk1/2, and PKA. These results suggested that endogenous Cdc42 could exert a negative regulatory influence on intrinsic migration/invasion and some potentially relevant changes in phosphorylation of PKCdelta, Erk1/2, and PKA of some aggressive breast cancer cells. PMID- 21618529 TI - Voltage sensitive phosphoinositide phosphatases of Xenopus: their tissue distribution and voltage dependence. AB - Voltage-sensitive phosphatases (VSPs) are unique proteins in which membrane potential controls enzyme activity. They are comprised of the voltage sensor domain of an ion channel coupled to a lipid phosphatase specific for phosphoinositides, and for ascidian and zebrafish VSPs, the phosphatase activity has been found to be activated by membrane depolarization. The physiological functions of these proteins are unknown, but their expression in testis and embryos suggests a role in fertilization or development. Here we investigate the expression pattern and voltage dependence of VSPs in two frog species, Xenopus laevis and Xenopus tropicalis, that are well suited for experimental studies of these possible functions. X. laevis has two VSP genes (Xl-VSP1 and Xl-VSP2), whereas X. tropicalis has only one gene (Xt-VSP). The highest expression of these genes was observed in testis, ovary, liver, and kidney. Our results show that while Xl-VSP2 activates only at positive membrane potentials outside of the physiological range, Xl-VSP1 and Xt-VSP phosphatase activity is regulated in the voltage range that regulates sperm-egg fusion at fertilization. PMID- 21618531 TI - Molecular chaperones as therapeutic targets to counteract proteostasis defects. AB - The health of cells is preserved by the levels and correct folding states of the proteome, which is generated and maintained by the proteostasis network, an integrated biological system consisting of several cytoprotective and degradative pathways. Indeed, the health conditions of the proteostasis network is a fundamental prerequisite to life as the inability to cope with the mismanagement of protein folding arising from genetic, epigenetic, and micro-environment stress appears to trigger a whole spectrum of unrelated diseases. Here we describe the potential functional role of the proteostasis network in tumor biology and in conformational diseases debating on how the signaling branches of this biological system may be manipulated to develop more efficacious and selective therapeutic strategies. We discuss the dual strategy of these processes in modulating the folding activity of molecular chaperones in order to counteract the antithetic proteostasis deficiencies occurring in cancer and loss/gain of function diseases. Finally, we provide perspectives on how to improve the outcome of these disorders by taking advantage of proteostasis modeling. PMID- 21618530 TI - Mechanisms controlling neurite outgrowth in a pheochromocytoma cell line: the role of TRPC channels. AB - Transient Receptor Potential Canonical (TRPC) channels are implicated in modulating neurite outgrowth. The expression pattern of TRPCs changes significantly during brain development, suggesting that fine-tuning TRPC expression may be important for orchestrating neuritogenesis. To study how alterations in the TRPC expression pattern affect neurite outgrowth, we used nerve growth factor (NGF)-differentiated rat pheochromocytoma 12 (PC12) cells, a model system for neuritogenesis. In PC12 cells, NGF markedly up-regulated TRPC1 and TRPC6 expression, but down-regulated TRPC5 expression while promoting neurite outgrowth. Overexpression of TRPC1 augmented, whereas TRPC5 overexpression decelerated NGF-induced neurite outgrowth. Conversely, shRNA-mediated knockdown of TRPC1 decreased, whereas shRNA-mediated knockdown of TRPC5 increased NGF induced neurite extension. Endogenous TRPC1 attenuated the anti-neuritogenic effect of overexpressed TRPC5 in part by forming the heteromeric TRPC1-TRPC5 channels. Previous reports suggested that TRPC6 may facilitate neurite outgrowth. However, we found that TRPC6 overexpression slowed down neuritogenesis, whereas dominant negative TRPC6 (DN-TRPC6) facilitated neurite outgrowth in NGF differentiated PC12 cells. Consistent with these findings, hyperforin, a neurite outgrowth promoting factor, decreased TRPC6 expression in NGF-differentiated PC12 cells. Using pharmacological and molecular biological approaches, we determined that NGF up-regulated TRPC1 and TRPC6 expression via a p75(NTR)-IKK(2)-dependent pathway that did not involve TrkA receptor signaling in PC12 cells. Similarly, NGF up-regulated TRPC1 and TRPC6 via an IKK(2) dependent pathway in primary cultured hippocampal neurons. Thus, our data suggest that a balance of TRPC1, TRPC5, and TRPC6 expression determines neurite extension rate in neural cells, with TRPC6 emerging as an NGF-dependent "molecular damper" maintaining a submaximal velocity of neurite extension. PMID- 21618532 TI - The nuclear form of glutathione peroxidase 4 is associated with sperm nuclear matrix and is required for proper paternal chromatin decondensation at fertilization. AB - The nuclear isoform of the selenoprotein Phospholipid Hydroperoxide Glutathione Peroxidase (nGPx4) is expressed in haploid male germ cells, contains several cysteines and is able to oxidize protein thiols, besides glutathione. In this study we have investigated the subnuclear localization of this isoform in isolated mouse male germ cells at different steps of maturation. Immunoblotting and confocal microscopy analyses of subnuclear fractions showed that nGPx4 is localized to the nuclear matrix together with well known markers of this subnuclear compartment like lamin B and topoisomerase IIbeta at all stages of germ cell differentiation. The peculiar nGPx4 distribution was confirmed by both biochemical and morphological analyses of COS-1 cells overexpressing Flag-tagged nGPx4. To test the functional role of nGPx4 in the process of chromatin assembly, sperm isolated from the caput and the cauda epididymides of wild-type (WT) and genetically deficient in nGPx4 (nGPx4-KO) mice were analyzed in an in vitro chromatin decondensation assay. Results showed that sperm from nGPx4-KO mice were more prone to decondense than those from WT mice at all stages of epididymal maturation, providing conclusive evidence that nGPx4 is required for a correct sperm chromatin compaction. We next addressed the issue of whether the lack of nGPx4 impacts on early events occurring at fertilization. Indeed, in vitro fertilization experiments showed an acceleration of sperm chromatin dispersion in oocytes fertilized by nGpx4-KO sperm compared with control. Overall these data indicate that the absence of nGPx4 leads to sperm nuclear matrix/chromatin instability that may negatively affect the embryo development. PMID- 21618533 TI - Hyposmotic stress causes ATP release and stimulates Na,K-ATPase activity in porcine lens. AB - Purinergic receptors in lens epithelium suggest lens function can be altered by chemical signals from aqueous humor or the lens itself. Here we show release of ATP by intact porcine lenses exposed to hyposmotic solution (200 mOsm). 18alpha glycyrrhetinic acid (AGA) added together with probenecid eliminated the ATP increase. N-ethylmaleimide (200 uM), an exocytotic inhibitor, had no significant effect on ATP increase. Lenses exposed to hyposmotic solution displayed a ~400% increase of propidium iodide (PI) entry into the epithelium. The increased ability of PI (MW 668) to enter the epithelium suggests possible opening of connexin and/or pannexin hemichannels. This is consistent with detection of connexin 43, connexin 50, and pannexin 1 in the epithelium and the ability of AGA + probenecid to prevent ATP release. Na,K-ATPase activity doubled in the epithelium of lenses exposed to hyposmotic solution. The increase of Na,K-ATPase activity did not occur when apyrase was used to prevent extracellular ATP accumulation or when AGA + probenecid prevented ATP release. The increase of Na,K ATPase activity was inhibited by the purinergic P2 antagonist reactive blue-2 and pertussis toxin, a G-protein inhibitor, but not by the P2X antagonist PPADS. Hyposmotic solution activated Src family kinase (SFK) in the epithelium, judged by Western blot. The SFK inhibitor PP2 abolished both SFK activation and the Na,K ATPase activity increase. In summary, hyposmotic shock-induced ATP release is sufficient to activate a purinergic receptor- and SFK-dependent mechanism that stimulates Na,K-ATPase activity. The responses might signify an autoregulatory loop initiated by mechanical stress or osmotic swelling. PMID- 21618534 TI - Anthrax lethal toxin disrupts the endothelial permeability barrier through blocking p38 signaling. AB - Exposure to anthrax causes life-threatening disease through the action of the toxin produced by the Bacillus anthracis bacteria. Lethal factor (LF), an anthrax toxin component which causes severe vascular leak and edema, is a protease which specifically degrades MAP kinase kinases (MKK). We have recently shown that p38 MAP kinase activation leading to HSP27 phosphorylation augments the endothelial permeability barrier. We now show that treatment of rat pulmonary microvascular endothelial cells with anthrax lethal toxin (LeTx), which is composed of LF and the protective antigen, increases endothelial barrier permeability and gap formation between endothelial cells through disrupting p38 signaling. LeTx treatment increases MKK3b degradation and in turn decreases p38 activity at baseline as well as after activation of p38 signaling. Consequently, LeTx treatment decreases activation of the p38 substrate kinase, MK2, and the phosphorylation of the latter's substrate, HSP27. LeTx treatment disrupts other signaling pathways leading to suppression of Erk-mediated signaling, but these effects do not correlate with LeTx-induced barrier compromise. Overexpressing phosphomimicking (pm)HSP27, which protects the endothelial permeability barrier against LeTx, blocks LeTx inactivation of p38 and MK2, but it does not block MKK3b degradation or Erk inactivation. Our results suggest that LeTx might cause vascular leak through inactivating p38-MK2-HSP27 signaling and that activating HSP27 phosphorylation specifically restores p38 signaling and blocks anthrax LeTx toxicity. The fact that barrier integrity could be restored by pmHSP27 overexpression without affecting degradation of MKK3b, or inactivation of Erk, suggests a specific and central role for p38-MK2-HSP27 in endothelial barrier permeability regulation. PMID- 21618535 TI - Hepatitis B virus X protein induces IKKalpha nuclear translocation via Akt dependent phosphorylation to promote the motility of hepatocarcinoma cells. AB - Hepatitis B virus (HBV) X protein (HBx) has been implicated in HBV-associated carcinogenesis through activation of IkappaB kinase (IKK)/nuclear factor kappa B (NF-kappaB) signaling pathway. Besides activating NF-kappaB in the cytoplasm, IKKalpha was found in the nucleus to regulate gene expression epigenetically in response to various stimuli. However, it is unknown whether nuclear IKKalpha plays a role in HBx-associated tumor progression. Moreover, the molecular mechanism underlying IKKalpha nuclear transport also remains to be elucidated. Here, we disclosed HBx as a new inducer of IKKalpha nuclear transport in hepatoma cells. HBx induced IKKalpha nuclear transport in an Akt-dependent manner. HBx activated Akt promoted IKKalpha nuclear translocation via phosphorylating its threonine-23 (Thr23). In addition, IKKalpha ubiquitination enhanced by HBx and Akt also contributed to the IKKalpha accumulation in the nucleus, indicating the involvement of ubiquitination in Akt-increased IKKalpha nuclear transport in response to HBx. Furthermore, inhibition of IKKalpha nuclear translocation by mutation of its nuclear localization signal and Thr23 diminished IKKalpha dependent cell migration. Taken together, our findings shed light on the molecular mechanism of IKKalpha nuclear translocation and provide a potential role of nuclear IKKalpha in HBx-mediated hepatocellular carcinoma (HCC) progression. PMID- 21618536 TI - Cooperation between Shh and IGF-I in promoting myogenic proliferation and differentiation via the MAPK/ERK and PI3K/Akt pathways requires Smo activity. AB - Sonic Hedgehog (Shh) has been shown to promote adult myoblast proliferation and differentiation and affect Akt phosphorylation via its effector Smoothened (Smo). Here, the relationship between Shh and insulin-like growth factor I (IGF-I) was examined with regard to myogenic differentiation via signaling pathways which regulate this process. Each factor enhanced Akt and MAPK/ERK (p42/44) phosphorylation and myogenic factor expression levels in a dose-responsive manner, while combinations of Shh and IGF-I showed additive effects. Blockage of the IGF-I effects by neutralizing antibody partially reduced Shh's effects on signaling pathways, suggesting that IGF-I enhances, but is not essential for Shh effects. Addition of cyclopamine, a Smo inhibitor, reduced Shh- and IGF-I-induced Akt phosphorylation in a similar manner, implying that Shh affects gain of the IGF-I signaling pathway. This implication was also examined via a genetic approach. In cultures derived from Smo(mut) (MCre;Smo(flox/flox)) mice lacking Smo expression specifically in hindlimb muscles, IGF-I-induced Akt and p42/44 phosphorylation was significantly reduced compared to IGF-I's effect on Smo(cont) cells. Moreover, remarkable inhibition of the stimulatory effect of IGF-I on myogenic differentiation was observed in Smo(mut) cultures, implying that intact Smo is required for IGF-I effects in myoblasts. Immunoprecipitation assays revealed that tyrosine-phosphorylated proteins, including the regulatory unit of PI3K (p85), are recruited to Smo in response to Shh. Moreover, IGF-IR was found to associate with Smo in response to Shh and to IGF-I, suggesting that Shh and IGF-I are already integrated at the receptor level, a mechanism by which their signaling pathways interact in augmenting their effects on adult myoblasts. PMID- 21618537 TI - In ovo feeding of IGF-1 to ducks influences neonatal skeletal muscle hypertrophy and muscle mass growth upon satellite cell activation. AB - To investigate reasons for the muscle increase observed when eggs are treated by IGF-1 and whether or not satellite cell activation is specific to different types of myofibers, duck eggs were administrated with IGF-1. After injection, during the neonatal stages, the duck breast muscle and leg muscle were isolated for analysis. The muscle weight, muscle fiber diameter (MFD), cross-sectional area (CSA), the number of myofibers per unit area (MFN) and frequency of satellite cell activation and mitosis at the embryo stage of 27 days (27E) and the postnatal stage of 2 days after hatching (P2D) were determined. In addition, expression of two important myogenic transcription factors MyoD and Myf5 were detected and compared in the two types of muscle tissues. Results indicated that IGF-1 administration increased the duck body weight, MFD, CSA, MFN, and quantity of activated satellite cells and mitotic nuclei in the two types of muscle tissues. The MyoD and Myf5 expressed at a higher level in IGF-1-treated muscle. IGF-1 stimulated muscle weight growth more in the leg muscle than in the breast muscle. These results indicate that in ovo feeding of IGF-1 can stimulate duck growth and, especially, lead to increased muscle hypertrophy. These increases appear to be mainly dependent on the activation of satellite cells, some of which proliferate and fuse to the myofiber, enabling increased muscle mass. IGF-1 can indirectly affect satellite cells by regulating the expression of two important myogenic transcription factors, MyoD and Myf5, which help activate satellite cells. PMID- 21618538 TI - Neuronal specific betaPix-b stimulates actin-dependent processes via the interaction between its PRD and WH1 domain of N-WASP. AB - betaPix, a Pak-interacting nucleotide exchange factor (Cool-1/p85SPR), is a Cdc42/Rac1-specific guanine nucleotide exchange factor (GEF) involved in various actin-related processes. Many previous studies have focused on ubiquitously expressed betaPix-a, while the role of the neuronal-specific isoform betaPix-b is still unknown, especially whether its role is distinct from or similar to betaPix a. Here we show that unlike betaPix-a, overexpression of betaPix-b stimulates actin-dependent comet formation in BHK21 cells. This effect is attributed to the interaction between its proline-rich domain (PRD) and the WH1 domain of N-WASP. In addition, we show that overexpression of betaPix-b stimulates actin-dependent dendritic spine formation in rat hippocampal neurons in culture, a formation that is blocked by co-expression of the WH1 domain of N-WASP or the PRD of betaPix-b. Knocking-down endogenous expression of betaPix-b by shRNA reduced the number of dendritic spines, which were rescued only by PRD-containing betaPix-b mutants. GEF activity of betaPix-b is also required for these effects. The results show that neuronal-specific betaPix-b stimulates actin-dependent processes in cells via the interaction between its PRD and the WH1 domain of N-WASP. Our results identify N-WASP as the first protein shown to interact with the PRD of betaPix-b, raising the possibility that, as an N-WASP WH1-binding protein, betaPix-b may regulate N-WASP's activity in cells. PMID- 21618540 TI - The chemokine CXCL12 and its receptor CXCR4 promote glioma stem cell-mediated VEGF production and tumour angiogenesis via PI3K/AKT signalling. AB - Chemokines and their receptors are actively involved in inflammation, immune responses, and cancer development. Here we report the detection of CD133(+) glioma stem-like cells (GSCs) co-expressing a chemokine receptor CXCR4 in human primary glioma tissues. These GSCs were located in areas adjacent to tumour vascular capillaries, suggesting an association between GSCs and tumour angiogenesis. To test this hypothesis, we isolated CD133(+) GSCs from surgical specimens of human primary gliomas and glioma cell lines. As compared to CD133(-) cells, CD133(+) GSCs expressed significantly higher levels of CXCR4 mRNA and protein, and migrated more efficiently in response to the CXCR4 ligand CXCL12. In addition, CXCL12 induced vascular endothelial growth factor (VEGF) production by CD133(+) GSCs via activation of the PI3K/AKT signalling pathway. Furthermore, knocking down of CXCR4 using RNA interference or inhibition of CXCR4 function by an antagonist AMD3100 not only reduced VEGF production by CD133(+) GSCs in vitro, but also attenuated the growth and angiogenesis of tumour xenografts in vivo formed by CD133(+) GSCs in SCID mice. These results indicate that CXCL12 and its receptor CXCR4 promote GSC-initiated glioma growth and angiogenesis by stimulating VEGF production. PMID- 21618541 TI - The MYB-NFIB gene fusion-a novel genetic link between adenoid cystic carcinoma and dermal cylindroma. AB - We have recently shown that the recurrent t(6;9)(q22 ~ 23;p23 ~ 24) translocation in adenoid cystic carcinoma (ACC) of the breast and head and neck results in a fusion of the two transcription factor genes MYB and NFIB. Here we demonstrate, for the first time, that benign sporadic, dermal cylindromas also express the MYB NFIB gene fusion. RT-PCR and immunohistochemical analyses revealed that eight of 12 analysed tumours (67%) expressed MYB-NFIB fusion transcripts and/or stained positive for MYB protein. Nucleotide sequence analyses confirmed that the composition of the chimeric transcript variants identified was identical to that in ACC, suggesting a similar molecular mechanism of activation of MYB in cylindroma as in ACC. In contrast, no evidence for the presence of the MYB-NFIB fusion was found in other types of basaloid skin and salivary gland tumours, indicating that the fusion indeed has a restricted expression pattern. Our findings broaden the spectrum of neoplasms associated with MYB oncogene activation and reveal a novel genetic link between ACC and dermal cylindroma. These results, together with our previous observations, further strengthen the evidence for common molecular pathways of importance for the development of both benign and malignant breast, salivary and adnexal tumours. PMID- 21618539 TI - Clozapine impairs insulin action by up-regulating Akt phosphorylation and Ped/Pea 15 protein abundance. AB - Clinical and experimental evidence indicates that atypical antipsychotics impair glucose metabolism. We investigated whether clozapine may directly affect insulin action by analyzing insulin signaling in vitro and in vivo. Clozapine reduced insulin-stimulated glucose uptake in PC12 and in L6 cells, representative models of neuron and skeletal muscle, respectively. Consistently, clozapine reduced insulin effect on insulin receptor (IR) by 40% and on IR substrate-1 (IRS1) tyrosine phosphorylation by 60%. Insulin-stimulated Akt phosphorylation was also reduced by about 40%. Moreover, insulin-dependent phosphorylation of protein kinase C-zeta (PKC-zeta) was completely blunted in clozapine-treated cells. Interestingly, clozapine treatment was accompanied by an insulin-independent increase of Akt phosphorylation, with no change of IR, IRS1, and PKC-zeta basal phosphorylation. The cellular abundance of Ped/Pea-15, an Akt substrate and inducer of insulin resistance, was also increased following clozapine exposure, both in the absence and in the presence of cyclohexymide, a protein synthesis inhibitor. Similar as in cellular models, in the caudate-putamen and in the tibialis muscle of clozapine-treated C57/BL/KsJ mice, Akt phosphorylation and Ped/Pea-15 protein levels were increased and PKC-zeta phosphorylation was decreased. Thus, in these experimental models, clozapine deranged Akt function and up-regulated Ped/Pea-15, thereby inhibiting insulin stimulation of PKC-zeta and of glucose uptake. PMID- 21618542 TI - The use of 13C-erythromycin as an in vivo probe to evaluate CYP3A-mediated drug interactions in rats. AB - (14)C-erythromycin breath test has been utilized to evaluate the extent of CYP3A activity in vivo. However, its radioactivity sometimes impedes its clinical application. In this study, we employed erythromycin labeled with (13)C ((13)C EM), a nonradioactive stable isotope, as an in vivo probe of breath test to evaluate CYP3A-mediated drug interactions in rats. A physiologically based pharmacokinetic (PBPK) model to describe (13)CO(2) exhalation altered by drug interactions was newly constructed. Rats received an intravenous or oral administration of (13)C-EM with or without a CYP3A inhibitor or inducer, that is, ketoconazole (KCZ) or dexamethasone (DEX), respectively. Breath samples were taken at designated times, measured with an infrared spectrophotometer, and the Delta(13) CO(2) value (0/00) in each sample was obtained. The C(max) and AUC(0-t) of Delta(13) CO(2) were significantly decreased with KCZ and increased with DEX. The PBPK model in this study successfully described the (13)CO(2) exhalation after (13)C-EM administration in the absence and presence of drug interactions. In conclusion, this study proposed a simple and rapid in vivo methodology to utilize (13)C-EM for the quantitative analysis of CYP3A inhibition and induction. This method using small animals may be useful in early drug development processes. PMID- 21618543 TI - Evaluation of transport mechanism of prodrugs and parent drugs formed by intracellular metabolism in Caco-2 cells with modified carboxylesterase activity: temocapril as a model case. AB - The intestinal absorption mechanism of temocapril, an ester-type prodrug of temocaprilat, was evaluated using Caco-2 cell monolayers with or without active carboxylesterase (CES)-mediated hydrolysis. The inhibition of CES-mediated hydrolysis was achieved by pretreatment of the monolayers with bis-p-nitrophenyl phosphate (BNPP), which inhibited 94% of the total hydrolysis of temocapril in the Caco-2 cells. The remaining 6% hydrolysis was due to the presence of serine esterases, other than CES, on the cell membranes. Transport experiments under CES inhibited conditions showed temocapril not to be a substrate for peptide transporter 1 (PEPT1) or organic anion transporting polypeptides (OATPs), but to be an inhibitor of PEPT1; P-glycoprotein (P-gp) and breast-cancer-resistant protein (BCRP) were responsible for the efflux of temocapril, which was mainly absorbed by passive diffusion at low apical pH. In Caco-2 cell monolayers with CES-mediated hydrolysis intact, temocaprilat derived from temocapril, was 2.5 fold more rapidly transported into the apical compartment than into the basolateral compartment due to the presence of microvilli on the apical membrane. In contrast, temocaprilat at low intracellular concentrations, was preferentially transported across the basolateral membrane under CES-inhibited conditions. PMID- 21618544 TI - Effects of uremic toxins on transport and metabolism of different biopharmaceutics drug disposition classification system xenobiotics. AB - Chronic kidney disease (CKD) is recognized to cause pharmacokinetic changes in renally excreted drugs; however, pharmacokinetic changes are also reported for drugs that are nonrenally eliminated. Few studies have investigated how uremic toxins may affect drug transporters and metabolizing enzymes and how these may result in pharmacokinetic/metabolic changes in CKD. Here, we investigated the effects of uremic toxins and human uremic serum on the transport of the prototypical transporter substrate [(3) H]-estrone sulfate and three Biopharmaceutics Drug Disposition Classification System (BDDCS) drugs, propranolol, losartan, and eprosartan. We observed a significant decrease in [(3) H]-estrone sulfate, losartan, and eprosartan uptake with some uremic toxins in both transfected cells and rat hepatocytes. The uptake of losartan was decreased in rat and human hepatocytes (28% and 48%, respectively) in the presence of hemodialysis (HD) serum. Time-course studies of losartan showed a 27%, 65%, and 68% increase in area under the curve (AUC) in the presence of HD serum, rifampin, and sulfaphenazole, respectively. Intracellular losartan AUC decreased significantly in the treatment groups, and the metabolite AUC decreased by 41% and 26% in rifampin- and sulfaphenazole-treated group, respectively. The intracellular AUC of eprosartan increased 190% in the presence of HD serum. These studies indicate that the uremic toxins contained in HD serum play an important role in drug disposition through drug transporters, and that there would be differential effects depending on the BDDCS classification of the drug. PMID- 21618545 TI - Effects of lard on the formation of volatiles from the Maillard reaction of cysteine with xylose. AB - BACKGROUND: The presence of lipid oxidation products in the Maillard reaction pathway is of particular interest today. The objective of this study was to investigate the effect of lard and its oxidation products on the formation of volatiles from cysteine and xylose model systems. RESULTS: Headspace volatiles generated in reaction mixtures were examined by solid-phase microextraction in combination with gas chromatography-mass spectrometry. The addition of lipid had a suppressing effect on most of the sulfur-containing compounds derived from the reaction between cysteine and xylose, especially for 2-methyl-3-furanthiol, 2 furanmethanethiol, 2-methylthiophene, and 3-methylthiophene. One of the intermediates-furfural-was also formed in much lower concentration when lard was present. In addition, cysteine and xylose modified lipid oxidation pathways, so that lipid-derived alcohols, alkylfurans and aliphatic acids were formed rather than aldehydes. Compared with the lard heated alone, most aldehydes were formed at lower levels in the lard-containing reaction mixtures, and several aldehydes including hexanal, heptanal, (2E)-heptenal and (2E,4E)-heptadienal were absent. CONCLUSION: The addition of lipid was inhibitory to the formation of most sulfur containing compounds in the Maillard reaction. Furthermore, Maillard reaction products influenced the formation of products from lipid oxidation. PMID- 21618546 TI - Influence of dietary supplementation with Bacillus-fermented adlay on lipid metabolism, antioxidant status and intestinal microflora in hamsters. AB - BACKGROUND: The effects of polished and dehulled Bacillus-fermented adlay on lipid metabolism, antioxidant status and intestinal microflora were examined in hyperlipidaemic hamsters fed a high-cholesterol diet. RESULTS: Hamsters administered Bacillus-fermented adlay experienced significantly reduced (P < 0.05) serum and hepatic total cholesterol (by 37-43% and 42-49% respectively) and triglyceride (by 22-27% and 30-35% respectively) levels compared with the high cholesterol group. Lower low-density lipoprotein cholesterol/high-density lipoprotein cholesterol ratios in serum and increased cholesterol (by 47-52%) and triglyceride (by 40-47%) contents in faeces were also observed. Bacillus fermented adlay lowered the levels of thiobarbituric acid-reactive substances, thus increasing total antioxidant and superoxide dismutase activities. In particular, polished Bacillus-fermented adlay had satisfactory antioxidant activity, similar to that of commercially available natto. Moreover, hamsters fed Bacillus-fermented adlay harboured greater populations of lactic acid bacteria, few coliforms and little Clostridium perfringens. CONCLUSION: This study has shown that changes in lipid metabolism, antioxidant status and intestinal microflora can be greatly modulated by Bacillus-fermented adlay, suggesting potential novel approaches to the treatment of primary cardiovascular and intestinal diseases. PMID- 21618547 TI - One-step multiplex PCR method for the determination of pecan and Brazil nut allergens in food products. AB - BACKGROUND: A one-step polymerase chain reaction (PCR) method for the simultaneous detection of the major allergens of pecan and Brazil nuts was developed. Primer pairs for the amplification of partial sequences of genes encoding the allergens were designed and tested for their specificity on a range of food components. RESULTS: The targeted amplicon size was 173 bp of Ber e 1 gene of Brazil nuts and 72 bp of vicilin-like seed storage protein gene in pecan nuts. The primer pair detecting the noncoding region of the chloroplast DNA was used as the internal control of amplification. The intrinsic detection limit of the PCR method was 100 pg mL(-1) pecan or Brazil nuts DNA. The practical detection limit was 0.1% w/w (1 g kg(-1)). The method was applied for the investigation of 63 samples with the declaration of pecans, Brazil nuts, other different nut species or nuts generally. In 15 food samples pecans and Brazil nuts allergens were identified in the conformity with the food declaration. CONCLUSION: The presented multiplex PCR method is specific enough and can be used as a fast approach for the detection of major allergens of pecan or Brazil nuts in food. PMID- 21618548 TI - Prediction and identification-based prediction of Chinese hepatitis C viral specific cytotoxic T lymphocyte epitopes. AB - Cytotoxic T lymphocytes (CTLs) play a critical role in the host immune response to infection by the Hepatitis C Virus (HCV). In the current study, a number of HCV CTL epitopes that represent the HLA polymorphisms found in the majority of Chinese people were predicted based on genomic and bioinformatic approaches. The predicted epitopes were evaluated for validity by examining the peptide-binding affinity for MHC class I molecules, the stability of peptide-MHC complexes, and frequencies of IFN gamma-positive T cells. Among the predicted epitope peptides, HLA-A2 restricted epitopes [NS4B (1793-1801) SMMAFSAAL] and HLA-B7 restricted epitopes [P7 (774-782) AAWYIKGRL] were able to induce high frequencies of IFN gamma-producing T cells, and the specific CTLs for other epitopes were not detected in peripheral blood lymphocytes from patients with HCV. Moreover, NS4B (1793-1801) exhibited high binding affinity for HLA-A2 molecules, and its stability of peptide-MHC class I complexes was sufficient, indicating that the high binding affinity for MHC class I molecules is an important factor for immunogenicity. Primary analyses of the immunogenicity of predicted epitopes, such as in the current study, will contribute to the future design of an efficient vaccine that will be able to induce vigorous, sustainable, and broad HCV-specific CTL responses for the Chinese population. PMID- 21618549 TI - Interruption of the circulation of an indigenous measles genotype and the introduction of other genotypes after a mass vaccination campaign in the Philippines. AB - Molecular analysis of measles viruses in the Philippines was conducted from 2000 to 2008. No confirmed measles cases were detected in the surveillance in 2005 after the mass vaccination campaign in 2004. However, a re-emergence of measles cases occurred in 2007, which was caused by other genotypes and the previous circulating genotype had disappeared. PMID- 21618550 TI - A prospective longitudinal study of BK virus infection in 120 Czech renal transplant recipients. AB - Polyomavirus BK (BKV) is a common human polyomavirus that rarely causes clinical symptoms in immunocompetent individuals. However, BK virus reactivation occurs in 20-40% of kidney transplant patients and 1-10% of cases present with BK virus associated nephropathy (BKVN) and reduced kidney allograft survival. In this study, 120 consecutive renal allograft recipients were monitored for BK virus replication by real-time PCR (qPCR) in the blood and urine during the first year post-transplantation and risk factors for BK viremia, viruria, and polyoma BKV associated nephropathy were evaluated. Receiver operating characteristic curve analysis was used to determine the cutoff points for assessing the risk of developing BKVN. In total, 1,243 samples were tested. BK-DNAuria >10(7) copies/ml and BK-DNAemia >10(4) copies/ml were found in 25.8% and 5% of the samples screened, respectively, during the 12 month follow-up period. BKVN was confirmed histologically in 3/120 patients and viremic patients were treated with dialysis for longer time periods and had higher levels of panel [corrected] reactive antibodies. Patients with viruria were also treated longer with dialysis and had impaired graft function 12 months post-transplantation. Patients with sustained viruria exhibited more acute rejection episodes than patients with transient viruria. Using receiver operating characteristic curve analysis, the cutoff point for viremia and viruria was redefined to 10(3) copies/ml serum for BK viremia and a cutoff point of 6.7 * 10(7) copies/ml in urine. In conclusion, polyoma BK viremia and viruria are frequent findings in kidney transplant recipients that warrant intensive monitoring as a means of preventing graft failure [corrected]. PMID- 21618551 TI - Human papillomavirus type distribution in vulval intraepithelial neoplasia determined using PapilloCheck DNA Microarray. AB - Vulval intraepithelial neoplasia is a precursor of vulval carcinoma, and is frequently associated with human papillomavirus (HPV) infection. Estimates of HPV prevalence in vulval intraepithelial neoplasia vary widely in the UK. The objective of this study was to assess HPV infection in a sample of women with vulval intraepithelial neoplasia, confirmed histologically, and determine the proportion of disease associated with HPV types targeted by prophylactic HPV vaccines. HPV infection was assessed in biopsies from 59 patients using the Greiner Bio-One PapilloCheck(r) DNA chip assay. Valid results were obtained for 54 cases. HPV infection was present in 43 of the 54 cases (79.6%: 95% CI 67.1 88.2%). The most common HPV types were HPV 16 (33/54: 61.1%), HPV 33 (8/54: 14.8%), HPV 6 (5/54: 9.3%), and HPV 42 (3/54: 5.6%). The mean age of HPV positive women was significantly less than the mean age of HPV negative women. This is the largest UK series of vulval intraepithelial neoplasia in which HPV type has been investigated, and 34/54 (63.0%, 95% CI: 49.6-78.6%) cases were associated with HPV 16/18, which are targeted by current prophylactic HPV vaccines. PMID- 21618552 TI - Symptomatic and asymptomatic infections of rotavirus, norovirus, and adenovirus among hospitalized children in Xi'an, China. AB - Rotavirus (RV), norovirus (NoV), and adenovirus (AdV) have been reported as the common viral pathogens of acute gastroenteritis in children. To determine the prevalence of RV, NoV, and AdV infections among hospitalized children with and without symptoms of acute gastroenteritis, fecal specimens, and data on clinical symptoms were collected from 201 children with diarrhea and 53 children without diarrhea admitted to the Xi'an Children's Hospital in Xi'an, China between March 2009 and May 2010. RV, NoV, and AdV were identified in 68.7% (138/201), 20.4% (41/201), and 5.0% (10/201), respectively, of children with diarrhea. These three viruses were also detected in 13.2% (7/53), 35.9% (19/53), and 9.4% (6/53), respectively, of children without diarrhea. Diarrheal children infected with RV alone showed the average severity score of 6.5, statistically significant higher than the average score of 5.3 in children with unidentifiable viruses. GII.3 and GII.4 were the only two NoV genotypes identified, and the GII.4 sequences were genetically close to GII.4 2006b cluster. These findings highlight the importance of NoV as a causative agent of pediatric diarrhea after RV based on the clinical and epidemiological characteristics of NoV infection, and particularly convey information of asymptomatic infections of enteric viruses in young children. PMID- 21618553 TI - Detection of Merkel cell polyomavirus on environmental surfaces. AB - The Merkel cell polyomavirus (MCPyV) is a human virus identified recently which is associated with the Merkel cell carcinoma. This virus is also detected frequently in the skin of healthy individuals. The presence of MCPyV has been investigated on environmental surfaces in contact with human skin. Various surfaces in four laboratories, public places, and individual homes were swabbed. Human DNA and MCPyV DNA were detected in swabs by real-time PCR. MCPyV DNA levels were measured before and after DNase treatment in a set of 12 MCPyV DNA-positive samples. A total of 60 environmental surface samples were collected. Fifty-one (85.0%) were positive for human DNA detection and 45 (75.0%) were positive for MCPyV DNA detection. All samples positive for MCPyV DNA were positive for human DNA detection. After DNase treatment, a 1.3 log decrease in MCPyV DNA level was observed indicating that about 5% of viral DNA is protected from DNase degradation and might be associated with infectious virus. These results indicate that MCPyV DNA may be detected on environmental surfaces in contact with human skin. Detection of viral DNA might reflect the presence of infectious viral particles and transmission from environmental source to humans cannot be ruled out. PMID- 21618554 TI - Distribution of human papillomavirus genotypes in Paraguayan women according to the severity of the cervical lesion. AB - The incidence of cervical cancer in Paraguay is among the highest in the world. This study aimed to determine the distribution of human papillomavirus (HPV) genotypes in Paraguayan women, according to the severity of the cervical lesion. This cross-sectional study included 207 women without a squamous intraepithelial lesion, 164 with low-grade squamous intraepithelial lesions, 74 with high-grade squamous intraepithelial lesions, and 41 with cervical cancer. Type-specific HPV was determined by the polymerase chain reaction with MY9/11 L1 and GP5+/GP6+ L1 primers, followed by restriction fragment length polymorphism and reverse line blotting hybridization, respectively. In total, 12 high-risk and 24 low-risk HPVs types were detected. HPV 16 was the most prevalent, followed by HPV 18 in cervical cancer (14.6%), HPV 31 in high-grade squamous intraepithelial lesions (14.9%), HPVs 58/42 in low-grade squamous intraepithelial lesions (9.1% each), and HPVs 31/58 (2.4% each) in women without squamous intraepithelial lesions. Among 285 positive samples, 24.2% harbored multiple HPV types, being this more prevalent in women with squamous intraepithelial lesions (30.8% in low-grade squamous intraepithelial lesions, 22.5% in high-grade squamous intraepithelial lesions, and 22.0% in cervical cancer) than in women without lesions (9.3%). The higher prevalence of HPV 16 and other high-risk HPVs in women both with and without cervical lesions may explain the high incidence of cervical cancer in Paraguay. This information may be of importance for local decision makers to improve prevention strategies. In addition, these results may be useful as baseline pre-vaccination data for a future virological surveillance in Paraguay. PMID- 21618555 TI - Prevalence and genotypes of human parechovirus in stool samples from hospitalized children in Shanghai, China, 2008 and 2009. AB - Human parechoviruses (HPeVs) are widespread pathogens belonging to the Picornavirus family. The aim of the present study was to assess the prevalence and genetic diversity of HPeV in Shanghai, China, during a HPeV screening program in 2008 and 2009. Of 300 stool samples from children under the age of 5 years with acute diarrhea seen at Children's Hospital, Fudan University, Shanghai, China, 165 (55%) were HPeV-positive. The median age of infected children was 3 months. The prevalence of HPeV was high (57%) in infants up to 2 years old but dropped to 30.4% in children between 2 and 5 years old. The prevalence did not differ by sex. Infections were present throughout the year but peaked in July and August. The most predominant genotype was HPeV1. Of the 139 strains, 4 were found in 9 samples: HPeV4 (n = 4), HPeV5 (n = 1), HPeV6 (n = 1), and HPeV8 (n= 3). This study provided useful data on the epidemiology of HPeV infection by documenting the distribution of genotypes, age of infection, and seasonal patterns in Shanghai, China. PMID- 21618556 TI - Spontaneous loss of hepatitis C virus RNA from serum is associated with genotype 1 and younger age at exposure. AB - A variety of factors have been associated with spontaneous loss of hepatitis C virus (HCV)-RNA from serum, including infecting HCV type, although results are conflicting. This study aimed to investigate further whether infecting HCV type was linked to spontaneous loss of HCV-RNA. Serum samples from 321 untreated HCV antibody positive patients presenting at the Hepatology clinic at Addenbrooke's Hospital, Cambridge between 2004 and 2007 were tested. These individuals were classified either as HCV antibody and HCV-RNA positive (viremic, n = 219) or HCV antibody positive and repeatedly HCV-RNA negative (non-viremic, n = 102). Infecting HCV type was identified by genotyping (viremic) or serotyping (non viremic). Binomial regression analysis investigated the independent effect of HCV type on spontaneous loss of HCV-RNA from serum by comparing the two groups. Ninety-one percent of patients were found to be either genotype 1 or genotype 3. The prevalence of type 1 infection was greater among non-viremic (64.5%) than viremic individuals (45%). After controlling for the effects of potential confounding factors, multivariable analyses showed that individuals with type 1 infections were more likely to be non-viremic than genotype 3 infections (RR = 2.07; 95% CI: 1.25, 3.43; P = 0.005). Individuals infected at an older age were also less likely to become HCV-RNA negative spontaneously (RR = 0.42 comparing those infected at >=20 years of age against those infected at <20 years of age, 95% CI: 0.25, 0.72; P = 0.002). In conclusion, the results suggest that HCV genotype 1 infections are more likely than genotype 3 infections to become spontaneously non-viremic, as are infections acquired at younger age. PMID- 21618557 TI - Clarifying and measuring filial concepts across five cultural groups. AB - Literature on responsibility of adult children for aging parents reflects lack of conceptual clarity. We examined filial concepts across five cultural groups: African-, Asian-, Euro-, Latino-, and Native Americans. Data were randomly divided for scale development (n = 285) and cross-validation (n = 284). Exploratory factor analysis on 59 items identified three filial concepts: Responsibility, Respect, and Care. Confirmatory factor analysis on a 12-item final scale showed data fit the three-factor model better than a single factor solution despite substantial correlations between the factors (.82, .82 for Care with Responsibility and Respect, and .74 for Responsibility with Respect). The scale can be used in cross-cultural research to test hypotheses that predict associations among filial values, filial caregiving, and caregiver health outcomes. PMID- 21618558 TI - Quantitative symmetry and chirality--a fast computational algorithm for large structures: proteins, macromolecules, nanotubes, and unit cells. AB - Symmetry is one of the most fundamental properties of nature and is used to understand and investigate physical properties. Classically, symmetry is treated as a binary qualitative property, although other physical properties are quantitative. Using the continuous symmetry measure (CSM) methodology one can quantify symmetry and correlate it quantitatively to physical, chemical, and biological properties. The exact analytical procedures for calculating the CSM are computationally expensive and the calculation time grows rapidly as the structure contains more atoms. In this article, we present a new method for calculating the CSM and the related continuous chirality measure (CCM) for large systems. The new method is much faster than the full analytical procedures and it reduces the calculation time dependency from N! to N(2), where N is the number of atoms in the structure. We evaluate the cost of the applied approximations, estimate the error of the method, and show that deviations from the analytical solutions are within an error of 2%, and in many cases even less. The method is applicable at the moment for the cyclic symmetry point groups- C(i), C(s), C(n), and S(n), and therefore it can be used also for chirality measures, which are the minimal of the S(n) measures. We demonstrate the application of the method for large structures across chemistry: proteins, macromolecules, nanotubes, and large unit cells of crystals. PMID- 21618559 TI - SDOCK: a global protein-protein docking program using stepwise force-field potentials. AB - Fast Fourier transform (FFT) method limits the forms of scoring functions in global protein-protein docking. On the other hand, force field potentials can effectively describe the energy hyper surface of biological macromolecules. In this study, we developed a new protein-protein docking program, SDOCK, that incorporates van der Waals attractive potential, geometric collision, screened electrostatic potential, and Lazaridis-Karplus desolvation energy into the scoring function in the global searching process. Stepwise potentials were generated from the corresponding continuous forms to treat the structure flexibility. After optimization of the atom solvation parameters and the weights of different potential terms based on a new docking test set that contains 142 cases with small or moderate conformational changes upon binding, SDOCK slightly outperformed the well-known FFT based global docking program ZDOCK3.0. Among the 142 cases tested, 52.8% gave at least one near-native solutions in the top 100 solutions. SDOCK was also tested on six blind testing cases in Critical Assessment of Predicted Interactions rounds 13 to 18. In all six cases, the near native solutions could be found within the top 350 solutions. Because the SDOCK approach performs global docking based on force-field potentials, one of its advantages is that it provides global binding free energy surface profiles for further analysis. The efficiency of the program is also comparable with that of other FFT based protein-protein docking programs. SDOCK is available for noncommercial applications at http://mdl.ipc.pku.edu.cn/cgi-bin/down.cgi. PMID- 21618560 TI - Fetal and neonatal alloimmune thrombocytopenia: prenatal interventions. AB - Fetal and neonatal alloimmune thrombocytopenia (FNAIT) is a potentially devastating condition, which may lead to intracranial haemorrhage (ICH) in the fetus or neonate, often with death or major neurological damage as consequence. In the absence of screening, preventive measures are only possible in the next pregnancy of women with an affected child. Controversy exists on the best intervention to minimise the risk of ICH. Most centres have abandoned treatment with serial fetal blood sampling (FBS) and platelet transfusions, because of a high rate of complications and the availability of quite effective non-invasive alternatives. In pregnancies with FNAIT and a previous affected child without ICH, weekly intravenous administration of immunoglobulins to the mother appears close to 100% effective to prevent fetal or neonatal ICH. Some centres add prednisone; this combination leads to slightly higher platelet counts at birth. In pregnant women with a previous child with ICH, the recurrence risk seems particularly high, and more aggressive maternal medical treatment is recommended, starting earlier with immunoglobulins. Whether a higher intravenous immunoglobulin dose or the addition of prednisone is really necessary is unclear. What does seem to be clear is that the use of FBS should be minimised, possibly even abandoned completely. PMID- 21618561 TI - Do repeated arrays of regulatory small-RNA genes elicit genomic imprinting?: Concurrent emergence of large clusters of small non-coding RNAs and genomic imprinting at four evolutionarily distinct eutherian chromosomal loci. AB - The basic premise of the host-defense theory is that genomic imprinting, the parent-of-origin expression of a subset of mammalian genes, derives from mechanisms originally dedicated to silencing repeated and retroviral-like sequences that deeply colonized mammalian genomes. We propose that large clusters of tandemly-repeated C/D-box small nucleolar RNAs (snoRNAs) or microRNAs represent a novel category of sequences recognized as "genomic parasites", contributing to the emergence of genomic imprinting in a subset of chromosomal regions that contain them. Such a view is supported by evidence derived from studies of the imprinted snoRNA- and/or miRNA-encoding Dlk1-Dio3, Snurf-Snrpn, Sfbmt2, and C19MC domains. While adding a new piece to the challenging puzzle of mammalian genome history, this hypothesis also reinforces the notion that dissecting the features and molecular mechanisms that discriminate between "foreign" and "endogenous" sequences is of crucial importance in the field of mammalian epigenetics. PMID- 21618562 TI - Development and validation of a highly sensitive LC-MS/MS method for quantitation of dexlansoprazole in human plasma: application to a human pharmacokinetic study. AB - A highly sensitive, specific and simple LC-MS/MS method was developed for the simultaneous estimation of dexlansoprazole (DEX) with 50 MUL of human plasma using omeprazole as an internal standard (IS). The API-4000 LC-MS/MS was operated under multiple reaction-monitoring mode using electrospray ionization. A simple liquid-liquid extraction process was used to extract DEX and IS from human plasma. The total run time was 2.00 min and the elution of DEX and IS occurred at 1.20 min. This was achieved with a mobile phase consisting of 0.2% ammonia acetonitrile (20:80, v/v) at a flow rate of 0.50 mL/min on an X-terra RP 18 (50 * 4.6 mm, 5 um) column. The developed method was validated in human plasma with a lower limit of quantitation of 2 ng/mL for DEX. A linear response function was established for the range of concentrations 2.00-2500.0 ng/mL (r > 0.998) for DEX. The intra- and inter-day precision values for DEX met the acceptance criteria as per FDA guidelines. DEX was stable in the battery of stability studies, viz. bench-top, auto-sampler and freeze-thaw cycles. The developed assay method was applied to an oral bioequivalence study in humans. PMID- 21618563 TI - In vivo metabolism study of polygalic acid in rat using HPLC-ESI-MSn. AB - A very simple and direct method has been established for the determination of polygalic acid and its metabolites in rat urine based on HPLC coupled with electrospray ionization multi-stage tandem mass spectrometry (HPLC-ESI-MS(n)). The rats were administered a single dose (100 mg/kg) of polygalic acid by oral gavage. The urine samples were collected and purified through a C(18) solid-phase extraction cartridge, and then these pretreated samples were injected into a reversed-phase C(18) column with a gradient elution program, whereas acetonitrile 0.5% aqueous formic acid was used as mobile phase and detected by an on-line MS/MS system. As a result, the parent drug and its four metabolites were identified and characterized in rat urine for the first time by comparing their changes in molecular mass (DeltaM), retention times and full-scan MS(n) spectra with those of the parent drug. A possible metabolic pathway of polygalic acid was investigated and proposed. More importantly, the results demonstrated that the newly developed method (HPLC-ESI-MS(n)) was sensitive, simple and suitable for the determination of polygalic acid and its metabolites in biological samples. PMID- 21618564 TI - A rapid LC-MS/MS method for quantitation of eszopiclone in human plasma: application to a human pharmacokinetic study. AB - A highly reproducible, specific and cost-effective LC-MS/MS method was developed for simultaneous estimation of eszopiclone (ESZ) with 50 MUL of human plasma using paroxetine as an internal standard (IS). The API-4000 LC-MS/MS was operated under the multiple reaction-monitoring mode using the electrospray ionization technique. A simple liquid-liquid extraction process was used to extract ESZ and IS from human plasma. The total run time was 1.5 min and the elution of ESZ and IS occurred at 0.90 min; this was achieved with a mobile phase consisting of 0.1% formic acid-methanol (15:85, v/v) at a flow rate of 0.50 mL/min on a Discover C(18) (50 * 4.6 mm, 5 um) column. The developed method was validated in human plasma with a lower limit of quantitation of 0.1 ng/mL for ESZ. A linear response function was established for the range of concentrations 0.10-120 ng/mL (r > 0.998) for ESZ. The intra- and inter-day precision values for ESZ were acceptable as per FDA guidelines. Eszopiclone was stable in the battery of stability studies, viz. bench-top, autosampler and freeze-thaw cycles. The developed assay method was applied to an oral bioequivalence study in humans. PMID- 21618565 TI - Elimination of hepatocellular carcinoma and acute hepatitis B in children 25 years after a hepatitis B newborn and catch-up immunization program. AB - Alaska Native people experience the highest rates of acute and chronic hepatitis B virus (HBV) infection and hepatocellular carcinoma (HCC) in the United States. We examined the effect of a universal newborn immunization with hepatitis B vaccine and mass population screening immunization program initiated in 1984 on rates of HBV and HCC in children 25 years later. During this time, the population of Alaska Native people grew from an estimated 75,000 to 130,000 persons. A surveillance system to detect acute HBV infection in Alaska Native facilities was established in 1981. Cases of HCC in children under 20 years of age were identified using a National Cancer Institute (NCI)-funded Cancer Registry established in 1969 coupled with an active surveillance program of screening persons with chronic HBV semiannually for alpha-fetoprotein since 1982. The incidence of acute symptomatic HBV infection in persons <20 years of age fell from cases 19/100,000 in 1981-1982 to 0/100,000 in 1993-1994. No cases of acute HBV have occurred in children since 1992. The incidence of HCC in persons <20 years decreased from 3/100,000 in 1984-1988 to zero in 1995-1999 and no cases have occurred since 1999. The number of identified hepatitis B surface antigen positive children <20 years in the Alaska Native population declined from 657 in 1987 to two in 2008. CONCLUSION: Universal newborn vaccination coupled with mass screening and immunization of susceptible Alaska Natives has eliminated HCC and acute symptomatic HBV infection among Alaska Native children and this approach is the best way to prevent HBV-related disease in children. PMID- 21618566 TI - Effect of telaprevir on the pharmacokinetics of cyclosporine and tacrolimus. AB - The hepatitis C virus protease inhibitor telaprevir is an inhibitor of the enzyme cytochrome P450 3A, responsible for the metabolism of both cyclosporine and tacrolimus. This Phase I, open-label, nonrandomized, single-sequence study assessed the effect of telaprevir coadministration on the pharmacokinetics of a single dose of either cyclosporine or tacrolimus in two separate panels of 10 healthy volunteers each. In Part A, cyclosporine was administered alone as a single 100-mg oral dose, followed by a minimum 8-day washout period, and subsequent coadministration of a single 10-mg oral dose of cyclosporine with either a single dose of telaprevir (750 mg) or with steady-state telaprevir (750 mg every 8 hours [q8h]). In Part B, tacrolimus was administered alone as a single 2-mg oral dose, followed by a minimum 14-day washout period, and subsequent coadministration of a single 0.5-mg dose of tacrolimus with steady-state telaprevir (750 mg q8h). Coadministration with steady-state telaprevir increased cyclosporine dose-normalized (DN) exposure (DN_AUC(0-infinity)) by approximately 4.6-fold and increased tacrolimus DN_AUC(0-infinity) by approximately 70-fold. Coadministration with telaprevir increased the terminal elimination half-life (t(1/2)) of cyclosporine from a mean (standard deviation [SD]) of 12 (1.67) hours to 42.1 (11.3) hours and t(1/2) of tacrolimus from a mean (SD) of 40.7 (5.85) hours to 196 (159) hours. CONCLUSION: In this study, telaprevir increased the blood concentrations of both cyclosporine and tacrolimus significantly, which could lead to serious or life-threatening adverse events. Telaprevir has not been studied in organ transplant patients; its use in these patients is not recommended because the required studies have not been completed to understand appropriate dose adjustments needed for safe coadministration of telaprevir with cyclosporine or tacrolimus, and regulatory approval has not been obtained. PMID- 21618568 TI - Mortality in nonalcoholic fatty liver disease: clues from the Cremona study. PMID- 21618569 TI - Postoperative transcatheter arterial chemoembolization should be recommended in the hepatocellular carcinoma treatment guidelines of the American Association for the Study of Liver Diseases. PMID- 21618570 TI - Vitamin D levels may explain the racial differences in response rates to antiviral therapy for chronic hepatitis C. PMID- 21618572 TI - Liver and serum iron: discrete regulators of hepatic hepcidin expression. PMID- 21618573 TI - c-Met represents a potential therapeutic target for personalized treatment in hepatocellular carcinoma. AB - c-Met, a high-affinity receptor for hepatocyte growth factor (HGF), plays a critical role in cancer growth, invasion, and metastasis. Hepatocellular carcinoma (HCC) patients with an active HGF/c-Met signaling pathway have a significantly worse prognosis. Although targeting the HGF/c-Met pathway has been proposed for the treatment of multiple cancers, the effect of c-Met inhibition in HCC remains unclear. The human HCC cell lines Huh7, Hep3B, MHCC97-L, and MHCC97-H were used in this study to investigate the effect of c-Met inhibition using the small molecule selective c-Met tyrosine kinase inhibitor PHA665752. MHCC97-L and MHCC97-H cells demonstrate a mesenchymal phenotype with decreased expression of E cadherin and increased expression of c-Met, fibronectin, and Zeb2 compared with Huh7 and Hep3B cells, which have an epithelial phenotype. PHA665752 treatment blocked phosphorylation of c-Met and downstream phosphoinositide 3-kinase/Akt and mitogen-activated protein kinase/Erk pathways, inhibited cell proliferation, and induced apoptosis in c-Met-positive MHCC97-L and MHCC97-H cells. In xenograft models, administration of PHA665752 significantly inhibited c-Met-positive MHCC97 L and MHCC97-H tumor growth, and PHA665752-treated tumors demonstrated marked reduction of both c-Met phosphorylation and cell proliferation. c-Met-negative Huh7 and Hep3B cells were not affected by c-Met inhibitor treatment in vitro or in vivo. In addition, c-Met-positive MHCC97-L and MHCC97-H cells demonstrated cancer stem cell-like characteristics, such as resistance to chemotherapy, tumor sphere formation, and increased expression of CD44 and ABCG2, and PHA665752 treatment suppressed tumor sphere formation and inhibited CD44 expression. CONCLUSION: c-Met represents a potential target of personalized treatment for HCC with an active HGF/c-Met pathway. PMID- 21618574 TI - Survival after yttrium-90 resin microsphere radioembolization of hepatocellular carcinoma across Barcelona clinic liver cancer stages: a European evaluation. AB - A multicenter analysis was conducted to evaluate the main prognostic factors driving survival after radioembolization using yttrium-90-labeled resin microspheres in patients with hepatocellular carcinoma at eight European centers. In total, 325 patients received a median activity of 1.6 GBq between September 2003 and December 2009, predominantly as whole-liver (45.2%) or right-lobe (38.5%) infusions. Typically, patients were Child-Pugh class A (82.5%), had underlying cirrhosis (78.5%), and had good Eastern Cooperative Oncology Group (ECOG) performance status (ECOG 0-1; 87.7%), but many had multinodular disease (75.9%) invading both lobes (53.1%) and/or portal vein occlusion (13.5% branch; 9.8% main). Over half had advanced Barcelona Clinic Liver Cancer (BCLC) staging (BCLC C, 56.3%) and one-quarter had intermediate staging (BCLC B, 26.8%). The median overall survival was 12.8 months (95% confidence interval, 10.9-15.7), which varied significantly by disease stage (BCLC A, 24.4 months [95% CI, 18.6 38.1 months]; BCLC B, 16.9 months [95% CI, 12.8-22.8 months]; BCLC C, 10.0 months [95% CI, 7.7-10.9 months]). Consistent with this finding , survival varied significantly by ECOG status, hepatic function (Child-Pugh class, ascites, and baseline total bilirubin), tumor burden (number of nodules, alpha-fetoprotein), and presence of extrahepatic disease. When considered within the framework of BCLC staging, variables reflecting tumor burden and liver function provided additional prognostic information. The most significant independent prognostic factors for survival upon multivariate analysis were ECOG status, tumor burden (nodules >5), international normalized ratio >1.2, and extrahepatic disease. Common adverse events were: fatigue, nausea/vomiting, and abdominal pain. Grade 3 or higher increases in bilirubin were reported in 5.8% of patients. All-cause mortality was 0.6% and 6.8% at 30 and 90 days, respectively. CONCLUSION: This analysis provides robust evidence of the survival achieved with radioembolization, including those with advanced disease and few treatment options. PMID- 21618575 TI - Diagnostic accuracy and reliability of ultrasonography for the detection of fatty liver: a meta-analysis. AB - Ultrasonography is a widely accessible imaging technique for the detection of fatty liver, but the reported accuracy and reliability have been inconsistent across studies. We aimed to perform a systematic review and meta-analysis of the diagnostic accuracy and reliability of ultrasonography for the detection of fatty liver. We used MEDLINE and Embase from October 1967 to March 2010. Studies that provided cross-tabulations of ultrasonography versus histology or standard imaging techniques, or that provided reliability data for ultrasonography, were included. Study variables were independently abstracted by three reviewers and double checked by one reviewer. Forty-nine (4720 participants) studies were included for the meta-analysis of diagnostic accuracy. The overall sensitivity, specificity, positive likelihood ratio, and negative likelihood ratio of ultrasound for the detection of moderate-severe fatty liver, compared to histology (gold standard), were 84.8% (95% confidence interval: 79.5-88.9), 93.6% (87.2-97.0), 13.3 (6.4-27.6), and 0.16 (0.12-0.22), respectively. The area under the summary receiving operating characteristics curve was 0.93 (0.91-0.95). Reliability of ultrasound for the detection of fatty liver showed kappa statistics ranging from 0.54 to 0.92 for intrarater reliability and from 0.44 to 1.00 for interrater reliability. Sensitivity and specificity of ultrasound was similar to that of other imaging techniques (i.e., computed tomography or magnetic resonance imaging). Statistical heterogeneity was present even after stratification for multiple clinically relevant characteristics. CONCLUSION: Ultrasonography allows for reliable and accurate detection of moderate-severe fatty liver, compared to histology. Because of its low cost, safety, and accessibility, ultrasound is likely the imaging technique of choice for screening for fatty liver in clinical and population settings. PMID- 21618576 TI - Impact of viral amino acid substitutions and host interleukin-28b polymorphism on replication and susceptibility to interferon of hepatitis C virus. AB - Amino acid (aa) substitutions of core 70 and 91 and in the NS5A (nonstructural protein 5A) interferon sensitivity determining region (ISDR) as well as genetic polymorphisms in the host interleukin-28B (IL28B) locus affect the outcome of interferon (IFN)-based therapies for patients with chronic hepatitis C. The combination of these factors and the quasispecies nature of the virus complicate understanding of the underlying mechanism. Using infectious hepatitis C virus (HCV) genotype 1b clone HCV-KT9, we introduced substitutions at both core aa70 (Arg to Gln) and aa91 (Leu to Met). We also introduced four and nine ISDR aa substitutions into core mutant HCV-KT9. Using human hepatocyte chimeric mice with different IL28B genotypes, we examined the infectivity, replication ability, and susceptibility to IFN of these clones. Although aa substitutions in the ISDR significantly impaired infectivity and replication ability of the virus, core aa70 and 91 substitutions did not. The effect of IFN treatment was similar in core wild-type and mutant viruses. Interestingly, virus titer was significantly higher in mice with the favorable IL28B allele (rs8099917 TT and rs12979860 CC) in the transplanted hepatocytes than in mice with hepatocytes from rs8099917 TG and rs12979860 TT donors (P < 0.001). However, the effect of IFN was significantly greater, and intrahepatic expression levels of IFN-stimulated genes were significantly higher in mice with the favorable IL28B allele. CONCLUSION: Our data suggest that HCV replication levels and response to IFN are affected by human hepatocyte IL28B single-nucleotide polymorphism genotype and mutations in the ISDR. The mechanism underlying the clinically observed association of wild type core protein in eradication-favorable host cells should be investigated further. PMID- 21618577 TI - Human hepatic cancer stem cells are characterized by common stemness traits and diverse oncogenic pathways. AB - Epigenetic mechanisms play critical roles in stem cell biology by maintaining pluripotency of stem cells and promoting differentiation of more mature derivatives. If similar mechanisms are relevant for the cancer stem cell (CSC) model, then epigenetic modulation might enrich the CSC population, thereby facilitating CSC isolation and rigorous evaluation. To test this hypothesis, primary human cancer cells and liver cancer cell lines were treated with zebularine (ZEB), a potent DNA methyltransferase-1 inhibitor, and putative CSCs were isolated using the side population (SP) approach. The CSC properties of ZEB treated and untreated subpopulations were tested using standard in vitro and in vivo assays. Whole transcriptome profiling of isolated CSCs was performed to generate CSC signatures. Clinical relevance of the CSC signatures was evaluated in diverse primary human cancers. Epigenetic modulation increased frequency of cells with CSC properties in the SP fraction isolated from human cancer cells as judged by self-renewal, superior tumor-initiating capacity in serial transplantations, and direct cell tracking experiments. Integrative transcriptome analysis revealed common traits enriched for stemness-associated genes, although each individual CSC gene expression signature exhibited activation of different oncogenic pathways (e.g., EGFR, SRC, and MYC). The common CSC signature was associated with malignant progression, which is enriched in poorly differentiated tumors, and was highly predictive of prognosis in liver and other cancers. CONCLUSION: Epigenetic modulation may provide a tool for prospective isolation and in-depth analysis of CSC. The liver CSC gene signatures are defined by a pernicious interaction of unique oncogene-specific and common stemness traits. These data should facilitate the identifications of therapeutic tools targeting both unique and common features of CSCs. PMID- 21618578 TI - NOX1/nicotinamide adenine dinucleotide phosphate, reduced form (NADPH) oxidase promotes proliferation of stellate cells and aggravates liver fibrosis induced by bile duct ligation. AB - Among multiple isoforms of nicotinamide adenine dinucleotide phosphate, reduced form (NADPH) oxidase expressed in the liver, the phagocytic NOX2 isoform in hepatic stellate cells (HSCs) has been demonstrated to play a key role in liver fibrogenesis. The aim of this study was to clarify the role of NOX1, a nonphagocytic form of NADPH oxidase, in the development of fibrosis using Nox1 deficient mice (Nox1KO). Liver injury and fibrosis were induced by bile duct ligation (BDL) and carbon tetrachloride in Nox1KO and wildtype littermate mice (WT). Primary HSCs were isolated to characterize the NOX1-induced signaling cascade involved in liver fibrogenesis. Following BDL, a time-dependent increase in NOX1 messenger RNA (mRNA) was demonstrated in WT liver. Compared with those in WT, levels of collagen-1alpha mRNA and hydroxyproline were significantly suppressed in Nox1KO with a reduced number of activated HSCs and less severe fibrotic lesions. The expression levels of alpha-smooth muscle actin, a marker of HSCs activation, were similar in cultured HSCs isolated from both genotypes. However, cell proliferation was significantly attenuated in HSCs isolated from Nox1KO. In these cells, the expression of p27(kip1) , a cell cycle suppressor, was significantly up-regulated. Concomitantly, a significant reduction in phosphorylated forms of Akt and forkhead box O (FOXO) 4, a downstream effector of Akt that regulates the transcription of p27(kip1) gene, was demonstrated in Nox1KO. Finally, the level of the oxidized inactivated form of phosphatase and tensin homolog (PTEN), a negative regulator of PI3K/Akt pathway, was significantly attenuated in HSCs of Nox1KO. CONCLUSION: These findings indicate that reactive oxygen species derived from NOX1/NADPH oxidase oxidize and inactivate PTEN to positively regulate the Akt/FOXO4/p27(kip1) signaling pathway. NOX1 may thus promote proliferation of HSCs and accelerate the development of fibrosis following BDL-induced liver injury. PMID- 21618580 TI - Molecular imaging of hepatic stellate cell activity by visualization of hepatic integrin alphavbeta3 expression with SPECT in rat. AB - The key factors in the pathogenesis of liver fibrosis are the activation and proliferation of hepatic stellate cells (HSCs), which express integrin alphavbeta3 after activation. This study aimed to explore the potential of (99m)Tc-labeled cyclic arginine-glycine-aspartic acid pentapeptide (cRGD) as a single photon emission computed tomography (SPECT) radiotracer to image hepatic integrin alphavbeta3 expression to reflect HSC activity in fibrotic livers. Rat models of liver fibrosis caused by thioacetamide or carbon tetrachloride (CCl(4)) treatment were employed to examine the expression and distribution of integrin alphavbeta3 during fibrotic progression or regression. The binding activity of radiolabeled cRGD to integrin alphavbeta3 was assessed in liver sections. SPECT was performed to determine hepatic integrin alphavbeta3 expression in rats with different stages of liver fibrosis. Protein and messenger RNA (mRNA) levels of integrin alphav and beta3 subunits were increased with the progression of liver fibrosis and reduced with its regression. The cell type that expressed the majority of integrin alphavbeta3 in fibrotic livers was found to be activated HSCs. The cRGD binding to activated HSCs displayed a high receptor-coupling affinity and an abundant receptor capacity. Iodine-125 ((125)I)-labeled cRGD bound to fibrotic liver sections and the binding activity was the highest in advanced fibrosis. Intravenously administered carboxyfluorescein-labeled cRGD was accumulated in fibrotic liver, and the accumulation amount was increased with the progression and reduced with the regression of fibrosis. A SPECT imaging study with (99m)Tc-labeled cRGD as a tracer demonstrated that the radioactivity ratio of liver to heart increased progressively along with severity of hepatic fibrosis. CONCLUSION: Hepatic integrin alphavbeta3 expression in fibrotic liver reflects HSC activity and its imaging using (99m)Tc-labeled cRGD as a SPECT radiotracer may distinguish different stages of liver fibrosis in rats. PMID- 21618579 TI - Nuclear expression of S100A4 calcium-binding protein increases cholangiocarcinoma invasiveness and metastasization. AB - Cholangiocarcinoma (CCA) carries a severe prognosis because of its strong invasiveness and early metastasization. In several patients, otherwise eligible for surgical resection, micrometastasis are already present at the time of surgery. The mechanisms responsible for CCA invasiveness are unclear. S100A4, a member of the S100 family of small Ca(2+)-binding proteins, is expressed in mesenchymal cells, regulates cell motility in several cell types, and is expressed in some epithelial cancers. Thus, we aimed to study the role of S100A4 in CCA invasiveness and metastasization. The expression of S100A4 was studied by immunohistochemistry in 93 human liver samples of CCA patients undergoing surgical resection and correlated with metastases development (67 cases) and patient survival following surgery using log rank tests and multivariate analysis. S100A4 expression was studied in EGI-1 and TFK-1, human CCA cell lines with and without nuclear S100A4 expression, respectively. Metastatic properties of CCA cells were assessed by xenotransplantation in severe combined immunodeficiency (SCID) mice after transduction with lentiviral vectors encoding firefly luciferase gene. Proliferation, motility (wound healing), invasiveness (Boyden chamber), and metalloproteinases (MMPs) secretion were studied in CCA cells, with or without lentiviral silencing of S100A4. Nuclear expression of S100A4 by neoplastic ducts was a strong predictor of metastasization and reduced survival after resection (P < 0.01). EGI-1 CCA cells showed stronger metastatic properties than TFK-1 when xenotransplanted in SCID mice. S100A4-silenced EGI-1 cells showed significantly reduced motility, invasiveness, and MMP-9 secretion in vitro, without changes in cell proliferation. CONCLUSION: Nuclear S100A4 identifies a subset of CCA patients with a poor prognosis after surgical resection. Nuclear expression of S100A4 increases CCA cells invasiveness and metastasization, indicating that S100A4 may also represent a potential therapeutic target. PMID- 21618581 TI - Activity-dependent gamma-aminobutyric acid release controls brain cortical tissue slice metabolism. AB - Vigabatrin (gamma-vinyl-GABA) is an irreversible inhibitor of the enzyme gamma aminobutyric acid (GABA) transaminase. It has been shown to increase levels of GABA in brain and result in increased release of GABA from nonsynaptic sources following activation. Here, we use a guinea pig cortical tissue slice model to identify the metabolic sequelae of vigabatrin when incubated with tissue slices alone or when the tissue slices were activated by ligands with targeted activating mechanisms. We show that incubation of slices with AMPA, the group II metabotropic glutamate antagonist EGLU [(2S)-alpha-ethylglutamic acid], or the GABA(B) R antagonist CGP 52432 in the presence of vigabatrin produces very similar metabolic profiles, consistent with the large-scale turning off of metabolic activity. This effect is blocked by the GABA(Arho) antagonist TPMPA [(1,2,5,6-tetrahydropyridin-4-yl)methylphosphinic acid]. Taken together, these results suggest that GABA, released following activation, acts on extrasynaptic receptors consistent with GABA(Arho) and that these receptors act as a kind of "master switch" that is capable of turning off a range of differently induced activities. PMID- 21618582 TI - Schwann-like adult stem cells derived from bone marrow and adipose tissue express gamma-aminobutyric acid type B receptors. AB - gamma-Aminobutyric acid type B receptors (GABA-B) are expressed in glial cells of the central and peripheral nervous systems, and recent evidence has shown their importance in modulating physiological parameters of Schwann cell (SC). SC play essential roles in peripheral nerve regeneration, but several drawbacks prevent their use for nerve repair. Adult stem cells from adipose tissue (ASC) or bone marrow (BM-MSC) can be differentiated into an SC-like phenotype and used as SC replacements. The aim of this study was to investigate GABA-B receptor functional expression in differentiated stem cells by assessing the similarity to SC. By means of RT-PCR and Western blot methodologies, BM-MSC and ASC were found to express both GABA-B1 and GABA-B2 receptor subunits. The expression levels of GABA B1b and GABA-B2 receptors were influenced by SC-like differentiation, as shown by Western blot studies. GABA-B receptor stimulation with baclofen reduced the proliferation rate of SC and differentiated ASC (dASC) but not that of dBM-MSC. In conclusion, both of the subunits that assemble into a functional GABA-B receptor are present in differentiated stem cells. Furthermore, GABA-B receptors in dASC are functionally active, regulating a key process such as proliferation. The presence of functional GABA-B receptors on differentiated stem cells opens new opportunities for a possible pharmacological modulation of their physiology and phenotype. PMID- 21618583 TI - Alternations of 14-3-3 theta and beta protein levels in brain during experimental sepsis. AB - The 14-3-3 family members play a crucial role in the determination of cell fate, exerting their antiapoptotic activity through directly interfering with the critical function of the mitochondrial core proapoptotic machinery. Dimerization of 14-3-3 is vital for the interaction with many of its client proteins and is regulated by phosphorylation. In a previous study, we observed time-dependent neuronal apoptosis during sepsis. Therefore, in the present study, we sought to evaluate the expression of 14-3-3 theta and beta isoforms in septic brain and their association with apoptosis. Sepsis was induced by a CLP model in Wistar rats that were sacrificed at predefined time points. Flow cytometric analysis showed a sepsis-induced, time-dependent alteration of 14-3-3 theta and beta isoforms in both Neun(+) and GFAP(+) cells. 14-3-3 theta was linearly correlated with apoptosis, and stratified analysis for alive and apoptotic neuronal cells demonstrated a gradual down-regulation of theta isoform in alive neurons and astrocytes. The phospho-P38 (pP38) MAP kinase levels were altered in a time dependent manner during sepsis, presenting a peak at 6 hr post-CLP. A significant correlation between the two isoforms of 14-3-3 was observed in septic rats, with the theta isoform predominant at all time points. The hippocampus, Purkinje cells, and glia-like cells showed intense immunohistochemical reactivity for 14-3 3 theta isoform, whereas the choroid plexus showed constantly increased beta isoform expression. Our results showed that sepsis alters the expression of both 14-3-3 theta and beta isoforms in a time-, cell-, and topography-dependent manner. PMID- 21618584 TI - Activation of stress kinases in the brain of mucopolysaccharidosis IIIB mice. AB - The accumulation of heparan sulfate (HS) in lysosomes is the primary consequence of the enzyme defect (alpha-N-acetylglucosaminidase) in mucopolysaccharidosis type IIIB. This accumulation triggers a cascade of pathological events that progressively leads to CNS pathology. Here we examined the activation of the three major stress kinases in the neuronal tissue of a murine model of the disease. ERK1/2 was significantly higher in the cortex of 1-2-month-old affected animals compared with wild-type (Wt) littermates. Similarly, ERK1/2 was stimulated in neurons cultured from MPS IIIB mice. SAPK/JNK was also found to be activated in the cortex of 1-2-month-old affected animals compared with Wt subjects, and the same was found for cultured neurons. In contrast, the active form of p38MAPK was lower in the cortex of 1-month-old MPS IIIB mice compared with Wt animals, but no significant difference was found between the two p38MAPK analyzed in normal and affected neurons cultured in vitro. These data indicate the possible involvement of MAPK dysregulation in the early stage of MPS IIIB brain disease. PMID- 21618585 TI - A new interpretation of stomatogenesis in a peritrich ciliate: using Campanella umbellaria as a model system. AB - The process of stomatogenesis in peritrich ciliates is still incompletely understood. Previous studies on the stomatogenesis of four species of peritrichs, Telotrochidium sp., Carchesium polypinum, Opercularia coarctata, and Astylozoon pyriforme conflict with one another in some cases and omit details of events in others. We described the entire process of stomatogenesis in the peritrich ciliate Campanella umbellaria (C. umbellaria) using an improved method of staining with protargol. Our results disagree with some previous studies with regard to the formation of some rudimentary structures, reorganization of the parental haplokinety, formation of new germinal rows, and separation of daughter oral complexes. The pattern of stomatogenesis characteristic of peritrichs is compared to the stomatogenetic patterns of three other oligohymenophorean subclasses and a hypothesis about the evolution of stomatogenesis in the class Oligohymenophorea is offered. Details of stomatogenesis need to be described and verified in a greater variety of peritrichs to clarify possible differences between taxa and make it possible to relate stomatogenesis to evolution within the subclass Peritrichia. Ultrastructural studies are the next step in description of morphogenetic processes in peritrichs, and characteristics of C. umbellaria make it a useful model for this work. PMID- 21618586 TI - Incident cognitive impairment is elevated in the stroke belt: the REGARDS study. AB - OBJECTIVE: To determine whether incidence of impaired cognitive screening status is higher in the southern Stroke Belt region of the United States than in the remaining United States. METHODS: A national cohort of adults age >=45 years was recruited by the Reasons for Geographic and Racial Differences in Stroke (REGARDS) study from 2003 to 2007. Participants' global cognitive status was assessed annually by telephone with the Six-Item Screener (SIS) and every 2 years with fluency and recall tasks. Participants who reported no stroke history and who were cognitively intact at enrollment (SIS >4 of 6) were included (N = 23,913, including 56% women; 38% African Americans and 62% European Americans; 56% Stroke Belt residents and 44% from the remaining contiguous United States and the District of Columbia). Regional differences in incident cognitive impairment (SIS score <=4) were adjusted for age, sex, race, education, and time between first and last assessments. RESULTS: A total of 1,937 participants (8.1%) declined to an SIS score <=4 at their most recent assessment, over a mean of 4.1 (+/-1.6) years. Residents of the Stroke Belt had greater adjusted odds of incident cognitive impairment than non-Belt residents (odds ratio, 1.18; 95% confidence interval, 1.07-1.30). All demographic factors and time independently predicted impairment. INTERPRETATION: Regional disparities in cognitive decline mirror regional disparities in stroke mortality, suggesting shared risk factors for these adverse outcomes. Efforts to promote cerebrovascular and cognitive health should be directed to the Stroke Belt. PMID- 21618587 TI - Cisplatin treatment of primary and metastatic epithelial ovarian carcinomas generates residual cells with mesenchymal stem cell-like profile. AB - Epithelial mesenchymal transition (EMT) and cancer stem cells (CSC) have been associated with resistance to chemotherapy. Eighty percent of ovarian cancer patients initially respond to platinum-based combination therapy but most return with recurrence and ultimate demise. To better understand such chemoresistance we have assessed the potential role of EMT in tumor cells collected from advanced stage ovarian cancer patients and the ovarian cancer cell line OVCA 433 in response to cisplatin in vitro. We demonstrate that cisplatin-induced transition from epithelial to mesenchymal morphology in residual cancer cells correlated with reduced E-cadherin, and increased N-cadherin and vimentin expression. The mRNA expression of Snail, Slug, Twist, and MMP-2 were significantly enhanced in response to cisplatin and correlated with increased migration. This coincided with increased cell surface expression of CSC-like markers such as CD44, alpha2 integrin subunit, CD117, CD133, EpCAM, and the expression of stem cell factors Nanog and Oct-4. EMT and CSC-like changes in response to cisplatin correlated with enhanced activation of extracellular signal-regulated kinase (ERK)1/2. The selective MEK inhibitor U0126 inhibited ERK2 activation and partially suppressed cisplatin-induced EMT and CSC markers. In vivo xenotransplantation of cisplatin treated OVCA 433 cells in zebrafish embryos demonstrated significantly enhanced migration of cells compared to control untreated cells. U0126 inhibited cisplatin induced migration of cells in vivo, suggesting that ERK2 signaling is critical to cisplatin-induced EMT and CSC phenotypes, and that targeting ERK2 in the presence of cisplatin may reduce the burden of residual tumor, the ultimate cause of recurrence in ovarian cancer patients. PMID- 21618588 TI - The retinol signaling pathway in mouse pluripotent P19 cells. AB - atRA (all-trans-retinoic acid), the active metabolite of retinol (vitamin A), is essential for embryogenesis and maintenance of cellular phenotype in adults. Chemicals that interfere with the metabolism of retinol to atRA, therefore, are a human health concern. During development of a screen for disruptors of this signaling pathway, we investigated whether the mouse pluripotent P19 cell metabolizes retinol to atRA and thus can be used in a cell-based screen for disruptors of the pathway. We found that retinol induced the identical pattern of homeobox gene expression as atRA and its precursor, retinal. Retinol was 160-fold less potent than atRA as an inducer, however. In spite of its lower potency, increased Hoxa1 gene expression was detected 30 min after retinol exposure and increased 40-fold by 2 h. Rdh10 and Aldh1a2/Raldh2, which together convert retinol to atRA in the embryo, were the predominant alcohol and aldehyde dehydrogenases expressed in P19 cells. The cell expressed high mRNA levels of retinol binding proteins, Rbp1 and Rbp4, and the 13,14-dihydroretinol saturase, Retsat. It also expressed all Rar and Rxr isotypes, Crabp1&2, the three Cyp26 genes, and both beta-carotene-cleaving genes, Bcmo1 and Bco2. The basal expression levels and retinol responsiveness of 25 pathway-related genes were quantitated by RT-qPCR. A test of the Aldh1a2 inhibitor, citral, showed that the disruption of the pathway was easily detected and quantitated showing that the P19 cell provides an in vitro model system for identifying and exploring the mechanism of action of chemicals that interfere with this critical cellular pathway. PMID- 21618589 TI - Baicalein protects against doxorubicin-induced cardiotoxicity by attenuation of mitochondrial oxidant injury and JNK activation. AB - The cardiotoxicity of doxorubicin limits its clinical use in the treatment of a variety of malignancies. Previous studies suggest that doxorubicin-associated cardiotoxicity is mediated by reactive oxygen species (ROS)-induced apoptosis. We therefore investigated if baicalein, a natural antioxidant component of Scutellaria baicalensis, could attenuate ROS generation and cell death induced by doxorubicin. Using an established chick cardiomyocyte model, doxorubicin (10 uM) increased cell death in a concentration- and time-dependent manner. ROS generation was increased in a dose-response fashion and associated with loss of mitochondrial membrane potential. Doxorubicin also augmented DNA fragmentation and increased the phosphorylation of ROS-sensitive pro-apoptotic kinase c-Jun N terminal kinase (JNK). Adjunct treatment of baicalein (25 uM) and doxorubicin for 24 h significantly reduced both ROS generation (587 +/- 89 a.u. vs. 932 a.u. +/- 121 a.u., P < 0.01) and cell death (30.6 +/- 5.1% vs. 46.8 +/- 8.3%, P < 0.01). The dissipated mitochondrial potential and increased DNA fragmentation were also ameliorated. Along with the reduction of ROS and apoptosis, baicalein attenuated phosphorylation of JNK induced by doxorubicin (1.7 +/- 0.3 vs. 3.0 +/- 0.4-fold, P < 0.05). Co-treatment of cardiomyocytes with doxorubicin and JNK inhibitor SP600125 (10 uM; 24 h) reduced JNK phosphorylation and enhanced cell survival, suggesting that the baicalein protection against doxorubicin cardiotoxicity was mediated by JNK activation. Importantly, concurrent baicalein treatment did not interfere with the anti-proliferative effects of doxorubicin in human breast cancer MCF-7 cells. In conclusion, baicalein adjunct treatment confers anti apoptotic protection against doxorubicin-induced cardiotoxicity without compromising its anti-cancer efficacy. PMID- 21618590 TI - Hepassocin regulates cell proliferation of the human hepatic cells L02 and hepatocarcinoma cells through different mechanisms. AB - Hepassocin (HPS) is a specific mitogenic active factor for hepatocytes, and inhibits growth by overexpression in hepatocellular carcinoma (HCC) cells. However, the mechanism of HPS regulation on growth of liver-derived cells still remains largely unknown. In this study, we found that HPS was expressed and secreted into the extracellular medium in cultured L02 human hepatic cells; conditional medium of L02 cells promoted proliferation of L02 cells and this activity could be blocked by anti-HPS antibody. Moreover, we identified the presence of receptor for HPS on L02 cells and HepG2 human hepatoma cells. Overproduction of truncated HPS, which signal peptide was deleted, significantly inhibited the proliferation of HCC cells and induced cell cycle arrest. These findings suggest that HPS promotes hepatic cell line L02 cells proliferation via an autocrine mechanism and inhibits HCC cells proliferation by an intracrine pathway. PMID- 21618591 TI - Regulation and function of axon guidance and adhesion molecules during olfactory map formation. AB - The olfactory system presents a practical model for investigating basic mechanisms involved in patterning connections between peripheral sensory neurons and central targets. Our understanding of olfactory map formation was advanced greatly by the discovery of cAMP signaling as an important determinant of glomerular positioning in the olfactory bulb. Additionally, several cell adhesion molecules have been identified recently that are proposed to regulate homotypic interactions among projecting axons. From these studies a model has emerged to partially explain the wiring of axons from widely dispersed neuron populations in the nasal cavity to relatively stereotyped glomerular positions. These advances have revitalized interest in axon guidance molecules in establishing olfactory topography, but also open new questions regarding how these patterns of guidance cues are established and function, and what other pathways, such as glycosylation, might be involved. This review summarizes the current state of this field and the important molecules that impact on cAMP-dependent mechanism in olfactory axon guidance. PMID- 21618592 TI - Complement regulator factor H in multiple sclerosis. AB - A recent proteomic study published in this journal demonstrated lower cerebrospinal fluid (CSF) expression of factor H (fH), an important complement regulator, along with two other complement proteins, in active multiple sclerosis (MS) patients. We have previously demonstrated raised serum fH levels in MS and here, an extended analysis, quantifying fH in CSF, demonstrates no change in fH levels in active disease, but significantly raised levels in progressive disease. These findings support our previous work showing raised serum fH in patients with progressive MS, and our results predict that CSF fH levels will be raised rather than reduced in active disease. PMID- 21618593 TI - Human periodontal ligament fibroblasts stimulate osteoclastogenesis in response to compression force through TNF-alpha-mediated activation of CD4+ T cells. AB - Periodontal ligament fibroblasts (PLF) sense and respond to mechanical stimuli and participate in alveolar bone resorption during orthodontic treatments. This study examined how PLF influence osteoclastogenesis from bone marrow-derived macrophages (BMM) after application of tension or compression force. We also investigated whether lymphocytes could be a primary stimulator of osteoclastic activation during alveolar bone remodeling. We found that mechanical forces inhibited osteoclastic differentiation from BMM in co-cultures with PLF, with PLF producing predominantly osteoprotegerin (OPG) rather than receptor activator of nuclear factor-kappaB (NF-kappaB) ligand (RANKL). In particular, PLF increased the expression of tumor necrosis factor (TNF)-alpha in response to compression. Additional experiments showed the presence of CD4- and B220-positive cells with a subsequent increase in tartrate-resistant acid phosphatase (TRAP)-positive cells and RANKL expression only at the compression side of the force-subjected periodontal tissues. Exogenous TNF-alpha increased the number of TRAP-positive cells and pit formation in the co-cultures of BMM with Jurkat, but not with BJAB cells and this effect was almost completely inhibited by antibodies to TNF-alpha or TNF receptor. Collectively, the current findings suggest that PLF secrete relatively higher levels of TNF-alpha at the compression side than at the tension side and this imbalance leads to RANKL expression by activating CD4+ T cells, thereby facilitating bone resorption during orthodontic tooth movement. PMID- 21618594 TI - Metformin stimulates osteoprotegerin and reduces RANKL expression in osteoblasts and ovariectomized rats. AB - Anti-diabetic drug metformin has been shown to enhance osteoblasts differentiation and inhibit osteoclast differentiation in vitro and prevent bone loss in ovariectomized (OVX) rats. But the mechanisms through which metformin regulates osteoclastogensis are not known. Osteoprotegerin (OPG) and receptor activator of nuclear factor kappaB ligand (RANKL) are cytokines predominantly secreted by osteoblasts and play critical roles in the differentiation and function of osteoclasts. In this study, we demonstrated that metformin dose dependently stimulated OPG and reduced RANKL mRNA and protein expression in mouse calvarial osteoblasts and osteoblastic cell line MC3T3-E1. Inhibition of AMP activated protein kinase (AMPK) and CaM kinase kinase (CaMKK), two targets of metformin, suppressed endogenous and metformin-induced OPG secretion in osteoblasts. Moreover, supernatant of osteoblasts treated with metformin reduced formation of tartrate resistant acid phosphatase (TRAP)-positive multi-nucleated cells in Raw264.7 cells. Most importantly, metformin significantly increased total body bone mineral density, prevented bone loss and decreased TRAP-positive cells in OVX rats proximal tibiae, accompanied with an increase of OPG and decrease of RANKL expression. These in vivo and in vitro studies suggest that metformin reduces RANKL and stimulates OPG expression in osteoblasts, further inhibits osteoclast differentiation and prevents bone loss in OVX rats. PMID- 21618595 TI - HepaCAM induces G1 phase arrest and promotes c-Myc degradation in human renal cell carcinoma. AB - Hepatocyte cell adhesion molecule (hepaCAM) encodes a generally inactive phosphorylated glycoprotein which mediates cancer cell proliferation, migration, and differentiation. We have reported that hepaCAM is down-regulated in renal cell carcinoma (RCC) and takes responsibility of cell growth inhibition. However, the precise mechanisms of hepaCAM inhibits cell growth is still unknown. In this study, we demonstrated that re-expression of hepaCAM can cause an accumulation in G0/G1 phase in 786-0 cells. This reaction was accompanied by a substantial reduction of c-Myc expression through using an ectopic hepaCAM expression system. Furthermore, we found a comparable decrease in proliferation and G0/G1 accumulation of 786-0 and RC-2 cells after treatment with a small molecule c-Myc inhibitor, 10058-F4. This indicated that the down regulation of c-Myc was an essential process in controlling growth inhibitory actions of hepaCAM. Nevertheless, re-expression of hepaCAM results in apparent reduction of c-Myc protein with no corresponding reduction of c-Myc mRNA. This suggests that this reaction might take place at a post-transcriptional level rather than transcriptional one. Consistent with these findings, hepaCAM decreased c-Myc stability by increasing the proportion of c-Myc phosphorylation on T58 which can be abrogated by a proteasomal inhibitor (MG132). Thus, our research implies that the decrease in c-Myc protein expression, resulting from ectopic expression of hepaCAM, may contribute to the inhibition of proliferation in these cells. PMID- 21618596 TI - Recurring extracorporeal circuit clotting during continuous renal replacement therapy resolved after single-session therapeutic plasma exchange. AB - We report a case of a 17-year-old white male with multiple fractures and multiorgan failure who developed oliguric acute renal failure requiring continuous renal replacement therapy. Repeated clotting of the extracorporeal circuit (ECC) prevented delivery of a minimally acceptable dose of renal replacement therapy despite adequate anticoagulation and dialysis catheter exchanges. Evaluation for a primary hypercoagulable state was negative, but his fibrinogen was elevated (1,320 mg/dL, normal range: 150-400 mg/dL), which is likely induced by his severe inflammatory state. A single session of therapeutic plasma exchange (TPE) with albumin and normal saline replacement was performed with subsequent drop in fibrinogen to 615 mg/dL. No further episodes of premature ECC clotting occurred, suggesting plasma factor(s) removed may have contributed to the clinical hypercoagulable state. TPE may play an adjunctive role in select cases of recurrent ECC clotting refractory to current anticoagulation techniques. PMID- 21618597 TI - Comparison of hematopoietic progenitor cell collections using the COBE Spectra version 7 and Amicus version 3.1 for patients with AL amyloidosis. AB - To address concerns about infused fluid volume during HPC collections in patients with AL amyloidosis, our institution has used a 26:1 anticoagulant (AC) ratio on the COBE Spectra and on the Fenwal Amicus. In this study, in a cohort of AL amyloid patients, we compared the Amicus version 3.1 to the Spectra version 7 MNC collections with regard to infused fluid volume, CD34+ cell yield, lymphocyte yield, cross-cellular content, and adverse reactions. Both instruments used a 26:1 AC ratio but the Amicus delivered significantly less AC per procedure (Amicus 678 mL vs. Spectra 753 mL). With comparable baseline CD34+ cell counts (Amicus 33 cells/MUL vs. Spectra 27 cells/MUL); Amicus collected significantly more CD34+ cells (3.1 vs. 1.5 * 106/kg) and equivalent lymphocytes (18.7 vs. 14.5 * 109. Amicus collected significantly fewer WBC (51.8 vs. 72.7 * 109), granulocytes (15.1 vs. 27.5 * 109), and PLT (2.3 vs. 3.9 * 1011) per procedure with equivalent RBC content (26 vs. 30 mL). CD34+ cell (5.0 vs. 4.4 * 106/kg) and lymphocyte doses (32.7 vs. 33.9 * 109) were equivalent in infused products collected on the Amicus and Spectra but the frequency of high volume products was lower for Amicus. Frequency and severity of adverse reactions during collection and infusion were similar for both. In this group of AL amyloid patients, Amicus was superior to Spectra with regard to fluid infused, CD34+ cell yield, and cross cellular contamination with equivalent lymphocyte yield and reaction incidence. PMID- 21618598 TI - The effect of drilling and screw fixation of the growth plate--an experimental study in rabbits. AB - Injury of the growth plate is a specific problem in traumatology and can cause limb deformity and length discrepancy as a result of growth arrest. The purpose of this study was to evaluate alterations of the growth plate after artificially created injuries. A total of 14 New Zealand White rabbits were used for this experiment. The right and left ulna of each animal was used resulting in a total of 28 ulnae. In six animals drill holes were driven into the growth plate either from the distal/epiphyseal side or from the proximal/metaphyseal side of the physis. In six animals a fracture of the distal ulna corresponding to a Salter Harris fracture type IV was created. This fracture was fixed by screws from either the epiphyseal or the metaphyseal side. Two animals served as controls. Histologic and radiologic examinations were performed to evaluate the growth process at 1, 2, 4, and 12 weeks following surgery. Drilling or fixation of the growth plate from the metaphyseal side resulted in temporary growth disturbance which was compensated within a short time. In contrast fixation from the epiphyseal side caused severe growth disturbances. Based on our findings K-wires or screws should be inserted from the metaphyseal side and be placed in the center of the growth plate. PMID- 21618599 TI - Molecular cloning and characterization of two nicotinic acetylcholine receptor beta subunit genes from Apis cerana cerana. AB - Nicotinic acetylcholine receptors (nAChRs) mediate fast cholinergic synaptic transmission in the insect nervous system and are important targets for insecticides. In this study, we identified and characterized two novel beta subunit genes (Accbeta1 and Accbeta2) from Apis cerana cerana. Homology analysis indicated that Accbeta1 and Accbeta2 possess characteristics that are typical of nAChR subunits although Accbeta2 was distinct from Accbeta1 and the other nAChR subunits, due to its unusual transmembrane structure and uncommon exon-intron boundary within the genomic region encoding the TM1 transmembrane domain. Analysis of the 5' flanking regions indicated that Accbeta1 and Accbeta2 possess different regulatory elements, suggesting that the genes might exhibit various expression and regulatory patterns. RT-PCR analysis demonstrated that Accbeta2 was expressed at a much higher level than Accbeta1 in the tissues of adult bees. During development, Accbeta1 was highly expressed at the pupal stages, whereas Accbeta2 was abundantly expressed at the larval stages. Furthermore, Accbeta1 and Accbeta2 were both induced by exposure to various insecticides and environmental stresses although Accbeta2 was more responsive than Accbeta1. These results indicate that Accbeta1 and Accbeta2 may have distinct roles in insect growth and development and that they may belong to separate regulatory pathways involved in the response to insecticides and environmental stresses. This report is the first description of the differences between the nAChR beta subunit genes in the Chinese honey bee and establishes an initial foundation for further study. PMID- 21618600 TI - Evaluation of methods accounting for population structure with pedigree data and continuous outcomes. AB - Methods to account for population structure (PS) in genome-wide association studies have been well developed in samples of unrelated individuals, but when a sample is composed of families, the task of finding and accounting for PS is not as straight forward. Family-based tests that condition on parental genotypes or their sufficient statistics are immune to biases due to PS, but are known to have low power, particularly for unselected samples. Population-based approaches that use all available data are an attractive alternative, but the methods have not been evaluated for continuous outcomes when a sample has both family and PS. Therefore, we compare through simulation the performance of population-based regression models that account for family and PS with continuous outcomes using a range of family sizes and structures, including two and three generational families with admixed and discrete PS. We find that when computation time is a concern, the Dupuis et al. efficient score test performs very well. When computational time is not an issue, a linear mixed effects model adjusting for genetic principal components tends to have slightly better power than the score test and may be preferred. PMID- 21618601 TI - Uncovering the total heritability explained by all true susceptibility variants in a genome-wide association study. AB - Genome-wide association studies (GWAS) have become increasingly popular recently and contributed to the discovery of many susceptibility variants. However, a large proportion of the heritability still remained unexplained. This observation raises queries regarding the ability of GWAS to uncover the genetic basis of complex diseases. In this study, we propose a simple and fast statistical framework to estimate the total heritability explained by all true susceptibility variants in a GWAS. It is expected that many true risk variants will not be detected in a GWAS due to limited power. The proposed framework aims at recovering the "hidden" heritability. Importantly, only the summary z-statistics are required as input and no raw genotype data are needed. The strategy is to recover the true effect sizes from the observed z-statistics. The methodology does not rely on any distributional assumptions of the effect sizes of variants. Both binary and quantitative traits can be handled and covariates may be included. Population-based or family-based designs are allowed as long as the summary statistics are available. Simulations were conducted and showed satisfactory performance of the proposed approach. Application to real data (Crohn's disease, HDL, LDL, and triglycerides) reveals that at least around 10 20% of variance in liability or phenotype can be explained by GWAS panels. This translates to around 10-40% of the total heritability for the studied traits. PMID- 21618602 TI - Detecting genetic interactions for quantitative traits with U-statistics. AB - The genetic etiology of complex human diseases has been commonly viewed as a process that involves multiple genetic variants, environmental factors, as well as their interactions. Statistical approaches, such as the multifactor dimensionality reduction (MDR) and generalized MDR (GMDR), have recently been proposed to test the joint association of multiple genetic variants with either dichotomous or continuous traits. In this study, we propose a novel Forward U Test to evaluate the combined effect of multiple loci on quantitative traits with consideration of gene-gene/gene-environment interactions. In this new approach, a U-Statistic-based forward algorithm is first used to select potential disease susceptibility loci and then a weighted U-statistic is used to test the joint association of the selected loci with the disease. Through a simulation study, we found the Forward U-Test outperformed GMDR in terms of greater power. Aside from that, our approach is less computationally intensive, making it feasible for high dimensional gene-gene/gene-environment research. We illustrate our method with a real data application to nicotine dependence (ND), using three independent datasets from the Study of Addiction: Genetics and Environment. Our gene-gene interaction analysis of 155 SNPs in 67 candidate genes identified two SNPs, rs16969968 within gene CHRNA5 and rs1122530 within gene NTRK2, jointly associated with the level of ND (P-value = 5.31e-7). The association, which involves essential interaction, is replicated in two independent datasets with P-values of 1.08e-5 and 0.02, respectively. Our finding suggests that joint action may exist between the two gene products. PMID- 21618603 TI - Evidence for gene-environment interaction in a genome wide study of nonsyndromic cleft palate. AB - Nonsyndromic cleft palate (CP) is a common birth defect with a complex and heterogeneous etiology involving both genetic and environmental risk factors. We conducted a genome-wide association study (GWAS) using 550 case-parent trios, ascertained through a CP case collected in an international consortium. Family based association tests of single nucleotide polymorphisms (SNP) and three common maternal exposures (maternal smoking, alcohol consumption, and multivitamin supplementation) were used in a combined 2 df test for gene (G) and gene environment (G * E) interaction simultaneously, plus a separate 1 df test for G * E interaction alone. Conditional logistic regression models were used to estimate effects on risk to exposed and unexposed children. While no SNP achieved genome wide significance when considered alone, markers in several genes attained or approached genome-wide significance when G * E interaction was included. Among these, MLLT3 and SMC2 on chromosome 9 showed multiple SNPs resulting in an increased risk if the mother consumed alcohol during the peri-conceptual period (3 months prior to conception through the first trimester). TBK1 on chr. 12 and ZNF236 on chr. 18 showed multiple SNPs associated with higher risk of CP in the presence of maternal smoking. Additional evidence of reduced risk due to G * E interaction in the presence of multivitamin supplementation was observed for SNPs in BAALC on chr. 8. These results emphasize the need to consider G * E interaction when searching for genes influencing risk to complex and heterogeneous disorders, such as nonsyndromic CP. PMID- 21618604 TI - Optimum designs for next-generation sequencing to discover rare variants for common complex disease. AB - Recent advances in next-generation sequencing technologies make it affordable to search for rare and functional variants for common complex diseases systematically. We investigated strategies for enriching rare variants in the samples selected for sequencing so as to optimize the power for their discovery. In particular, we investigated the roles of alternative sources of enrichment in families through computer simulations. We showed that linkage information, extreme phenotype, and nonrandom ascertainment, such as multiply affected families, constitute different sources for enriching rare and functional variants in a sequencing study design. Linkage is well known to have limited power for detecting small genetic effects, and hence not considered to be a powerful tool for discovering variants for common complex diseases. However, those families with some degree of family-specific linkage evidence provide an effective sampling strategy to sub-select the most linkage-informative families for sequencing. Compared with selecting subjects with extreme phenotypes, linkage evidence performs better with larger families, while extreme-phenotype method is more efficient with smaller families. Families with multiple affected siblings were found to provide the largest enrichment of rare variants. Finally, we showed that combined strategies, such as selecting linkage-informative families from multiply affected families, provide much higher enrichment of rare functional variants than either strategy alone. PMID- 21618605 TI - Improved genetic association tests for an ordinal outcome representing the disease progression process. AB - We are interested in detecting genetic variants that influence transition between discrete stages of a disease progression process, such as the natural history of progression to cervical cancer with the following four stages: (1) normal-human papillomavirus (HPV) exposed, (2) persistent infection with oncogenic HPV, (3) cervical intraepithelial neoplasia grades 2 or 3 (CIN2/3), and (4) cervical cancer. Standard statistical tests derived from the proportional odds model or polytomous regression model can be used to study this type of ordinal outcome. But these methods are either too sensitive to the proportion odds assumption or fail to take advantage of the restriction on the parameter space for the genetic variants. Two alternative tests, the maximum score test (MAX) and the adaptive P value combination test (Adapt-P), are proposed with the aim of striking a balance between efficiency and robustness. A simulation study demonstrates that MAX and Adapt-P have the most robust performance among all considered tests under various realistic scenarios. As a demonstration, we applied the considered tests to a genetic association study of cervical cancer. PMID- 21618606 TI - Evaluation of polygenic risk scores for predicting breast and prostate cancer risk. AB - Recently, polygenic risk scores (PRS) have been shown to be associated with certain complex diseases. The approach has been based on the contribution of counting multiple alleles associated with disease across independent loci, without requiring compelling evidence that every locus had already achieved definitive genome-wide statistical significance. Whether PRS assist in the prediction of risk of common cancers is unknown. We built PRS from lists of genetic markers prioritized by their association with breast cancer (BCa) or prostate cancer (PCa) in a training data set and evaluated whether these scores could improve current genetic prediction of these specific cancers in independent test samples. We used genome-wide association data on 1,145 BCa cases and 1,142 controls from the Nurses' Health Study and 1,164 PCa cases and 1,113 controls from the Prostate Lung Colorectal and Ovarian Cancer Screening Trial. Ten-fold cross validation was used to build and evaluate PRS with 10 to 60,000 independent single nucleotide polymorphisms (SNPs). For both BCa and PCa, the models that included only published risk alleles maximized the cross-validation estimate of the area under the ROC curve (0.53 for breast and 0.57 for prostate). We found no significant evidence that PRS using common variants improved risk prediction for BCa and PCa over replicated SNP scores. PMID- 21618607 TI - Combined R2* and diffusion tensor imaging changes in the substantia nigra in Parkinson's disease. AB - Recent magnetic resonance imaging studies suggest an increased transverse relaxation rate and reduced diffusion tensor imaging fractional anisotropy values in the substantia nigra in Parkinson's disease. The transverse relaxation rate and fractional anisotropy changes may reflect different aspects of Parkinson's disease-related pathological processes (ie, tissue iron deposition and microstructure disorganization). This study investigated the combined changes of transverse relaxation rate and fractional anisotropy in the substantia nigra in Parkinson's disease. High-resolution magnetic resonance imaging (T2-weighted, T2*, and diffusion tensor imaging) were obtained from 16 Parkinson's disease patients and 16 controls. Bilateral substantia nigras were delineated manually on T2-weighted images and coregistered to transverse relaxation rate and fractional anisotropy maps. The mean transverse relaxation rate and fractional anisotropy values in each substantia nigra were then calculated and compared between Parkinson's disease subjects and controls. Logistic regression, followed by receiver operating characteristic curve analysis, was employed to investigate the sensitivity and specificity of the combined measures for differentiating Parkinson's disease subjects from controls. Compared with controls, Parkinson's disease subjects demonstrated increased transverse relaxation rate (P<.0001) and reduced fractional anisotropy (P=.0365) in the substantia nigra. There was no significant correlation between transverse relaxation rate and fractional anisotropy values. Logistic regression analyses indicated that the combined use of transverse relaxation rate and fractional anisotropy values provides excellent discrimination between Parkinson's disease subjects and controls (c statistic=0.996) compared with transverse relaxation rate (c-statistic=0.930) or fractional anisotropy (c-statistic=0.742) alone. This study shows that the combined use of transverse relaxation rate and fractional anisotropy measures in the substantia nigra of Parkinson's disease enhances sensitivity and specificity in differentiating Parkinson's disease from controls. Further studies are warranted to evaluate the pathophysiological correlations of these magnetic resonance imaging measurements and their effectiveness in assisting in diagnosing Parkinson's disease and following its progression. PMID- 21618608 TI - A reappraisal of long-latency abdominal muscle reflexes in patients with propriospinal myoclonus. AB - BACKGROUND: We report 3 patients with typical clinical and electrophysiological characteristics of propriospinal myoclonus propagating from a thoracic spine generator. METHODS: In these patients, the pattern of recruitment of long-latency electromyographic reflexes in abdominal muscles was studied in response to various stimuli. RESULTS: Abdominal reflex latency varied from 60 to 140 ms depending on stimulus location. Latency increased from magnetic stimulation of the thoracic spine to electrical stimulation of the supraorbital nerve, electrical stimulation of the median nerve, and magnetic stimulation of the motor cortex. CONCLUSIONS: Long-latency abdominal reflex jerks are probably controlled by the brain stem to propriospinal system projections in patients with propriospinal myoclonus. The stereotyped pattern of recruitment of these reflexes could be of clinical utility to differentiate organic propriospinal myoclonus from psychogenic or mimicked jerks. PMID- 21618609 TI - Electrogastrographyc activity in Parkinson's disease patients with and without motor fluctuations. AB - BACKGROUND: Gastroenteric dysfunctions are very common in Parkinson's disease, but their relationship with dopaminergic response and motor fluctuations is still unclear. Electrogastrography is a noninvasive method for measuring gastric myoelectrical activity. METHODS: We evaluated the effects of levodopa intake on the motility of empty stomachs in Parkinson's disease patients with and without motor fluctuations. RESULTS: The electrogastrography findings showed a normal pattern not influenced by levodopa intake, unrelated to plasmatic levodopa concentrations and to clinical parameters. CONCLUSIONS: Our results suggest that at rest gastric activity of Parkinson's disease patients is normal and plasmatic levodopa variability is not influenced by gastric motility. PMID- 21618610 TI - Late-onset asymmetric myoclonus: an emerging syndrome. AB - BACKGROUND: Asymmetric cortical myoclonus is typically thought to be associated with either contralateral cortical structural lesions or degenerative disorders such as corticobasal degeneration when onset is in middle-aged or aged adults. This view has been challenged after a recent case series brought to light a syndrome of senile-onset, asymmetric cortical myoclonus not associated with any such identifiable disorders, thus, named "primary progressive myoclonus of aging." This is rare and no other reports have been published; hence, further such cases need to be highlighted. CASE REPORTS: Here, we describe 3 patients with some similarities, namely, adult-onset, asymmetric myoclonus that is most likely to be cortical, with an unremarkable thorough diagnostic workup, but with younger age at onset and longer follow-up time. CONCLUSIONS: This report expands on previous phenotypical descriptions attempting to further develop and refine this possible diagnostic entity. PMID- 21618611 TI - Lysosomal storage disorders and Parkinson's disease: Gaucher disease and beyond. AB - Parkinson's disease is associated with mutations in the glucocerebrosidase gene, which result in the enzyme deficiency causing Gaucher disease, the most common lysosomal storage disorder. We have performed an exhaustive literature search and found that additional lysosomal storage disorders might be associated with Parkinson's disease, based on case reports, the appearance of pathological features such as alpha-synuclein deposits in the brain, and substantia nigra pathology. Our findings suggest that the search for biochemical and cellular pathways that link Parkinson's disease with lysosomal storage disorders should not be limited exclusively to changes that occur in Gaucher disease, such as changes in glucocerebrosidase activity or in glucosylceramide levels, but rather include changes that might be common to a wide variety of lysosomal storage disorders. Moreover, we propose that additional genetic, epidemiological, and clinical studies should be performed to check the precise incidence of mutations in genes encoding lysosomal proteins in patients displaying Parkinson's symptoms. PMID- 21618612 TI - A preliminary study of finasteride in Tourette syndrome. PMID- 21618613 TI - Paranodal permeability in "myelin mutants". AB - Fluorescent dextran tracers of varying sizes have been used to assess paranodal permeability in myelinated sciatic nerve fibers from control and three "myelin mutant" mice, Caspr-null, cst-null, and shaking. We demonstrate that in all of these the paranode is permeable to small tracers (3 kDa and 10 kDa), which penetrate most fibers, and to larger tracers (40 kDa and 70 kDa), which penetrate far fewer fibers and move shorter distances over longer periods of time. Despite gross diminution in transverse bands (TBs) in the Caspr-null and cst-null mice, the permeability of their paranodal junctions is equivalent to that in controls. Thus, deficiency of TBs in these mutants does not increase the permeability of their paranodal junctions to the dextrans we used, moving from the perinodal space through the paranode to the internodal periaxonal space. In addition, we show that the shaking mice, which have thinner myelin and shorter paranodes, show increased permeability to the same tracers despite the presence of TBs. We conclude that the extent of penetration of these tracers does not depend on the presence or absence of TBs but does depend on the length of the paranode and, in turn, on the length of "pathway 3," the helical extracellular pathway that passes through the paranode parallel to the lateral edge of the myelin sheath. PMID- 21618614 TI - A quantitative measure of chirality inside nucleic acid databank. AB - We show the capability of a chirality index (Pietropaolo et al., Proteins 2008;70:667-677) to investigate nucleic acid structures because of its high sensitivity to helical conformations. By analyzing selected structures of DNA and RNA, we have found that sequences rich in cytosine and guanine have a tendency to left-handed chirality, in contrast to regions rich in adenine or thymine which show strong negative, right-handed, chirality values. We also analyze RNA structures, where specific loops and hairpin motifs are characterized by a well defined chirality value. We find that in nucleosome the chirality is exalted, whereas in ribosome it is reduced. Our results illustrate the sensitivity of this descriptor for nucleic acid conformations. PMID- 21618615 TI - Effect of fenoterol stereochemistry on the beta2 adrenergic receptor system: ligand-directed chiral recognition. AB - The beta(2) adrenergic receptor (beta(2)-AR) is a model system for studying the ligand recognition process in G protein-coupled receptors. Fenoterol (FEN) is a beta(2)-AR selective agonist that has two centers of chirality and exists as four stereoisomers. Radioligand binding studies determined that stereochemistry greatly influences the binding affinity. Subsequent Van't Hoff analysis shows very different thermodynamics of binding depending on the stereoconfiguration of the molecule. The binding of (S,x')-isomers is almost entirely enthalpy controlled whereas binding of (R,x')-isomers is purely entropy driven. Stereochemistry of FEN molecule also affects the coupling of the receptor to different G proteins. In a rat cardiomyocyte contractility model, (R,R')-FEN was shown to selectively activate G(s) protein signaling while the (S,R')-isomer activated both G(i) and G(s) protein. The overall data demonstrate that the chirality at the two chiral centers of the FEN molecule influences the magnitude of binding affinity, thermodynamics of local interactions within the binding site, and the global mechanism of beta(2)-AR activation. Differences in thermodynamic parameters and nonuniform G-protein coupling suggest a mechanism of chiral recognition in which observed enantioselectivities arise from the interaction of the (R,x')-FEN stereoisomers with a different receptor conformation than the one with which the (S,x')-isomer interacts. PMID- 21618616 TI - Fibrous architectures and organogels of tris(phenylethynylphenyl)adamantane molecules with amino acid moieties: their solvato-controlled helicity induction. AB - Non-disk-shaped molecules self-assembled to form columnar-type helical aggregates with gel ability through pi-stacking interactions among the central tris(phenylethynylphenyl)adamantane moieties, hydrogen-bonding interactions among the alanine parts with amide groups, and van der Waals interactions among the alkyl groups in nonpolar n-alkanes. The structural analyses of fibrous architectures with helicity were examined by FTIR, UV-Vis, and circular dichroism (CD) spectroscopy and FE-SEM measurements. In contrast, the formation of fibrous structures was observed from adamantane-based tripodal molecules due to solvophobic interactions in polar EtOH, which showed no detectable Cotton effect in the CD measurement, indicating the induction of solvato-controlled helicity in the self-assembly process. PMID- 21618620 TI - High fidelity self-recognition of isomeric oligopyridines in binary 2D self assembly and its application for separation. AB - Self-assembly in two binary mixtures based on three isomeric oligopyridines at the liquid/HOPG (highly oriented pyrolytic graphite) interface is presented. Despite their structural similarity the molecules display exclusive phase separation, which is attributed to the highly specific intermolecular hydrogen bonding interactions. Variation of the mole fractions in solution reveal strongly preferred adsorption of the major compound, which underlines the importance of self-recognition for self-assembly. Those findings at the molecular level can be applied to separation issues on a macroscopic scale, leading to a completely new concept of separation, which could have a strong impact on various chromatographic processes. PMID- 21618617 TI - Directed blood vessel growth using an angiogenic microfiber/microparticle composite patch. PMID- 21618621 TI - Mechanistic insights into ring-closing enyne metathesis with the second generation Grubbs-Hoveyda catalyst: a DFT study. AB - The full catalytic process (precatalyst activation, propagating cycle and active species interconversion) of the ring-closing enyne metathesis (RCEYM) reaction of 1-allyloxy-2-propyne with the Grubbs-Hoveyda complex as catalyst was studied by B3LYP density functional theory. Both the ene-then-yne and yne-then-ene pathways are considered and, for the productive catalytic cycle, the feasibility of the endo-yne-then-ene route is also explored. Calculations predict that the ene-then yne and yne-then-ene pathways proceed through equivalent steps, the only major difference being the order in which they take place. In this way, all alkene metathesis processes studied here involve four steps: olefin coordination, cycloaddition, cycloreversion and olefin decoordination. Among them, the two more energetically demanding ones are the olefin coordination and decoordination steps. The reaction of the alkyne fragment consists of two steps: alkyne coordination and alkyne skeletal reorganization, the latter of which has the highest Gibbs energy barrier. Comparison between the ene-then-yne and yne-then ene pathways shows that there is no clear energetic preference for either of the two processes, and thus both should be operative when unsubstituted enynes are involved. In addition, although the endo orientation is computed to be slightly disfavored, it is not ruled out for 1-allyloxy-2-propyne, and thus calculations seem to indicate that the exo versus endo selectivity is strongly influenced by the presence of substituents in the reagent. PMID- 21618622 TI - Development of a flexible strategy towards FR900482 and the mitomycins. AB - FR900482 and the mitomycins are two intriguing classes of alkaloid natural products that have analogous biological mechanisms and obvious structural similarity. Both classes possess potent anticancer activity, a feature that has led to their investigation and implementation for the clinical treatment of human cancer. Given the structural similarity between these natural products, we envisioned a common synthetic strategy by which both classes could be targeted through assembling the mitomycin skeleton prior to further oxidative functionalization. Realization of this strategy with respect to FR900482 was accomplished through the synthesis of 7-epi-FR900482, which displayed equal potency relative to the natural product against two human cancer cell lines. With the challenging goal of a synthesis of either mitomycin or FR900482 in mind, several methodologies were explored. While not all of these methods ultimately proved useful for our synthetic goal, a number of them led to intriguing findings that provide a more complete understanding of several methodologies. In particular, amination via pi-allyl palladium complexes for the synthesis of tetrahydroquinolines, eight-membered heterocycle formation via carbonylative lactamization, and amination through late-stage C-H insertion via rhodium catalysis all featured prominently in our synthetic studies. PMID- 21618624 TI - Roimatacene: an antibiotic against Gram-negative bacteria isolated from Cystobacter ferrugineus Cb G35 (Myxobacteria). AB - Roimatacene (1) was isolated from the myxobacterium Cystobacter ferrugineus strain Cb G35 in a bioactivity-guided process, by following the antimicrobial activity against Escherichia coli. Since 1 was extremely sensitive to oxygen, a protective isolation and handling protocol was developed, by utilizing the free radical scavenger 4-ethoxyphenol. The structure of 1 was determined by HRMS, 1D and 2D NMR spectroscopy and chemical derivatization to acetonides and Mosher esters to finally establish the absolute configuration. Methionine and acetate were identified as building blocks in the biosynthesis of 1 by feeding experiments with differently (13)C-labeled precursors. The antimicrobial activity of 1 was determined in a broad screening revealing 1 to inhibit several Gram negative bacteria. PMID- 21618623 TI - The crystal structure of the CeNA:RNA hybrid ce(GCGTAGCG):r(CGCUACGC). AB - Cyclohexenyl nucleic acids (CeNA) are characterised by the carbon-carbon double bond replacing the O4'-oxygen atom of the natural D-2'-deoxyribose sugar ring in DNA. CeNAs exhibit a high conformational flexibility, are stable against nuclease activity and their hybridisation is RNA selective. Additionally, CeNA has been shown to induce an enhanced biological activity when incorporated in siRNA. This makes CeNA a good candidate for siRNA and synthetic aptamer applications. The crystal structure of the synthetic CeNA:RNA hybrid ce(GCGTAGCG):r(CGCUACGC) has been solved with a resolution of 2.50 A. The CeNA:RNA duplex adopts an anti parallel, right-handed double helix with standard Watson-Crick base pairing. Analyses of the helical parameters revealed the octamer to form an A-like double helix. The cyclohexenyl rings mainly adopt the (3)H(2) conformation, which resembles the C3'-endo conformation of RNA ribose ring. This C3'-endo ring puckering was found in most of the RNA residues and is typical for A-family helices. The crystal structure is stabilised by the presence of hexahydrated magnesium ions. The fact that the CeNA:RNA hybrid adopts an A-type double helical conformation confirms the high potential of CeNAs for the construction of efficient siRNAs which can be used for therapeutical applications. PMID- 21618625 TI - Light-induced water oxidation by a Ru complex containing a bio-inspired ligand. AB - The new Ru complex 8 containing the bio-inspired ligand 7 was successfully synthesized and characterized. Complex 8 efficiently catalyzes water oxidation using Ce(IV) and Ru(III) as chemical oxidants. More importantly, this complex has a sufficiently low overpotential to utilize ruthenium polypyridyl-type complexes as photosensitizers. PMID- 21618626 TI - Hydrothermal synthesis of Na(0.5)La(0.5)TiO3-LaCrO3 solid-solution single-crystal nanocubes for visible-light-driven photocatalytic H2 evolution. AB - Cubic perovskite structure photocatalysts of Na(0.5)La(0.5)TiO(3) and (Na(0.5)La(0.5)TiO(3))(1.00)(LaCrO(3))(0.08) solid solution that consisted of well-defined single-crystal nanocubes were successfully prepared by means of facile and surfactant-free hydrothermal reactions for the first time. The results from different instrumental characterizations and theoretical calculations consistently confirmed the formation of nanocubic single-crystal solid solution of (Na(0.5)La(0.5)TiO(3))(1.00)(LaCrO(3))(0.08), and clearly revealed the modification of its physicochemical properties compared with those of Na(0.5)La(0.5)TiO(3). In particular, the effective narrowing of the bandgap (from 3.19 to 2.25 eV) by Cr(3+) in the solid solution made it possible to utilize visible light. The solid-solution configuration maintained the charge balance to preserve the valence of Cr(3+) rather than Cr(6+), and accommodated Cr(3+) with high content to form new energy bands instead of localized impurity levels. The hydrothermal preparation strategy ensured the formation of single crystals with high purity, few defects, and regulated morphology; it also guaranteed the valences of Ti(4+) and Cr(3+) in the solid solution. Consequently, the recombination of photogenerated carriers could be effectively suppressed to benefit photocatalytic H(2) evolution. (Na(0.5)La(0.5)TiO(3))(1.00)(LaCrO(3))(0.08) nanocubic single-crystal solid solution showed stable photocatalytic activity, and thus was proved to be a promising candidate for visible-light-driven photocatalytic H(2) evolution. PMID- 21618627 TI - Distinctly different chemical functionalities on the internal and the external surfaces of silica nanotubes, and their applications as multi-chemosensors. PMID- 21618628 TI - Push-pull amino succinimidyl ester thiophene-based fluorescent dyes: synthesis and optical characterization. AB - The design and synthesis of new fluorescent dyes with emission range at 490-650 nm are described. Their structural and electronic properties have been characterized by both experimental techniques and quantum-chemical calculations. The chromophores are donor-pi-bridge-acceptor push-pull compounds with a pi bridge of phenyl and thiophene rings and their combination. Compared with previous thiophene fluorophores, these dyes show significant redshift in the absorption and emission spectra and offer compact, red-emitting fluorophores. The dyes have amino succinimidyl active ester and can be readily conjugated to proteins, polymers and other amino-group-containing materials. PMID- 21618629 TI - Self-assembly into spheres of a hybrid diphenylalanine-porphyrin: increased fluorescence lifetime and conserved electronic properties. AB - A series of protected phenylalanine and diphenylalanine derivatives have been coupled through a peptide bond to a monoaminoporphyrin to form new materials. A comparative study in solution and in the solid state has been performed and confirmed new and interesting properties for the self-assembled hybrid materials while conserving the electronic properties of the chromophore. Thus, they are powerful candidates for use in dye-sensitized solar cells. PMID- 21618630 TI - Synthesis of 5-acetyloxazoles and 1,2-diketones from beta-alkoxy-beta ketoenamides and their subsequent transformations. AB - Lithiated alkoxyallenes, nitriles, and carboxylic acids have been employed as precursors in a three-component reaction leading to highly substituted beta alkoxy-beta-ketoenamides. Upon treatment with trifluoroacetic acid, these enamides could be easily cyclized to 5-acetyloxazole derivatives. The synthesis is very flexible with respect to the substitution pattern at C-2 and C-4 of the oxazole core. A mechanistic suggestion for the oxazole formation is presented on the basis of (18)O-labeled compounds and their mass spectrometric analysis. In several cases, 1,2-diketones are formed as side products or even as major components. The acetyl moiety at C-5 of the oxazole derivatives can efficiently be converted into alkenyl or alkynyl moieties, which allows a multitude of subsequent reactions. Condensation reactions of the acetyl group provided the expected oxime or hydrazone. By applying a Fischer reaction, the phenylhydrazone could be transferred into an indole, which emphasizes the potential of 5 acetyloxazoles for the preparation of highly substituted (poly)heterocyclic systems. The alkynyl group at C-2 is prone to addition reactions, providing an enamine with interesting photophysical properties. Sonogashira couplings were performed with 5-alkynyl-substituted oxazoles, furnishing the expected aryl substituted products. This alkynyl unit was employed for the preparation of a new, star-shaped trisoxazole derivative. The ability of this multivalent compound to form self-assembled monolayers between the basal plane of highly oriented pyrolytic graphite and 1-phenyloctane was demonstrated by scanning tunneling microscopy (STM). The star-shaped compound seems to prefer the C(3)-symmetric arrangement in this two-dimensional crystal. Two 1,2-diketones were smoothly converted into functionalized quinoxaline derivatives. PMID- 21618631 TI - Frequency-domain Fourier-transform terahertz spectroscopy of the single-molecule magnet (NEt4)[Mn2(5-Brsalen)2(MeOH)2Cr(CN)6]. AB - We have investigated the novel single-molecule magnet (NEt(4))[Mn(2)(5 Brsalen)(2)(MeOH)(2)Cr(CN)(6)] (1; 5-Brsalen = N,N'-ethylenebis(5 bromosalicylidene)iminato anion) using spectroscopic as well as magnetization and susceptibility measurements. Frequency-domain Fourier-transform terahertz electron paramagnetic resonance (FDFT THz-EPR) based on the generation of THz radiation from a synchrotron in combination with inelastic neutron scattering (INS) allows for the discrimination between intermultiplet and intramultiplet transitions. Together with ac/dc magnetic susceptibility measurements the obtained set of data provides a complete characterization of the lowest energetic magnetic excitations. We find that the new compound 1 exhibits much weaker intermolecular interactions than found in the closely related compound: K[Mn(2)(5 Brsalen)(2)(H(2)O)(2)Cr(CN)(6)] (2). Furthermore, two phonon lines in the vicinity of the magnetic excitations are detected. PMID- 21618632 TI - Lighting-up single-walled carbon nanotubes with silver nanoclusters. PMID- 21618633 TI - High-performance single-crystal-based organic field-effect transistors from pi extended porphyrin derivatives. PMID- 21618634 TI - Asymmetric formal [3+2] cycloaddition reaction of isocyanoesters to 2 oxobutenoate esters by a multifunctional chiral silver catalyst. PMID- 21618635 TI - S(N)1-type reactions in the presence of water: indium(III)-promoted highly enantioselective organocatalytic propargylation of aldehydes. PMID- 21618636 TI - Ruthenium catalysts containing rigid chiral diamines and achiral diphosphanes for highly enantioselective hydrogenation of aromatic ketones. PMID- 21618637 TI - Cooperative supramolecular polymerization and amplification of chirality in C3 symmetrical OPE-based trisamides. PMID- 21618638 TI - Concise synthesis of the antidepressive drug candidate GSK1360707 by a highly enantioselective gold-catalyzed enyne cycloisomerization reaction. PMID- 21618639 TI - Clinical assessment of a supplement of Pycnogenol(r) and L-arginine in Japanese patients with mild to moderate erectile dysfunction. AB - A double-blind parallel group comparison design clinical study was conducted in Japanese patients with mild to moderate erectile dysfunction to investigate the efficacy of a supplement containing Pycnogenol(r) and L-arginine. Subjects were instructed to take a supplement (Pycnogenol(r) 60 mg/day, L-arginine 690 mg/day and aspartic acid 552 mg/day) or an identical placebo for 8 weeks, and the results were assessed using the five-item erectile domain (IIEF-5) of the International Index of Erectile Function. Additionally, blood biochemistry, urinalysis and salivary testosterone were measured. Eight weeks of supplement intake improved the total score of the IIEF-5. In particular, a marked improvement was observed in 'hardness of erection' and 'satisfaction with sexual intercourse'. A decrease in blood pressure, aspartate transaminase and gamma glutamyl transpeptidase (gamma-GTP), and a slight increase in salivary testosterone were observed in the supplement group. No adverse reactions were observed during the study period. In conclusion, Pycnogenol(r) in combination with L-arginine as a dietary supplement is effective and safe in Japanese patients with mild to moderate erectile dysfunction. PMID- 21618640 TI - Temporal factors control hippocampal contributions to fear renewal after extinction. AB - Fear can be extinguished by repeated exposure to a cue that signals threat. However, extinction does not erase fear, as an extinguished cue presented in a context distinct from that of extinction results in renewed fear of that cue. The hippocampus, which is involved in the formation of contextual representations, is a natural candidate structure for investigations into the neural circuitry underlying fear renewal. Thus far, studies examining the necessity of the hippocampus for fear renewal have produced mixed results. We isolated the conditions under which the hippocampus may be required for renewal. Rats received lesions of the dorsal hippocampus either prior to tone fear conditioning or following extinction. Fear renewal was measured using discrete tone presentations or a long, continuous tone. The topography of fear responding at test was assessed by comparing "early" and "sustained" renewal, where early fear was determined by freezing to the first discrete tone or the equivalent initial segment of a continuous tone and sustained fear was determined by freezing averaged across all discrete tones or the entire continuous tone. We found that following pretraining damage of the hippocampus, early renewal remained intact regardless of lesion condition. However, sustained renewal only persisted in discrete, but not continuous, tone-tested animals. A more extensive analysis of the topography of fear responding revealed that the disruption of renewal was generated when the tone duration at test began to violate that used during extinction, suggesting that the hippocampus is sensitive to mismatches in CS duration. Postextinction lesions resulted in an overall reduction of fear renewal. This pattern of results is consistent with those observed for contextual fear conditioning, wherein animals display a resistance to anterograde amnesia despite the presence of a strong retrograde amnesia for the same contextual information. Furthermore, the data support a role for the hippocampus in sustaining renewal when the CS duration at test does not match that used during extinction. PMID- 21618641 TI - Proteins linked to spatial memory formation of CD1 mice in the multiple T-maze. AB - In own previous work CD1 mice were tested in the Multiple T-maze (MTM), a robust land maze allowing determination of latency to reach the goal box with food reward and to evaluate correct decisions made on the way to the goal box. Herein, hippocampi of these animals were used for the current study with the aim to investigate differences in protein levels between trained and yoked mice and, moreover, to determine differences in protein levels between trained and yoked mice with and without memory formation in the MTM. Three training sessions were carried out for four training days each, followed by probe trials on Days 5 and 12. Good and no-performers in the MTM were separated based on means and median of latency to reach the goal box on probe trial Day 12. Six hours following the probe trial on Day 12, animals were sacrificed and hippocampi were taken. Proteins were extracted and run on two-dimensional gel electrophoresis, spots were quantified and differentially expressed proteins were identified by mass spectrometry using an ion trap. Levels of 17 proteins were significantly different in trained vs. yoked mice. Seven proteins were differentially expressed comparing trained vs. yoked mice from good and no-performers. A series of proteins were significantly correlated with latency and may link these proteins to spatial memory formation. Differential protein expression in trained vs. yoked mice and in good and no-performers may allow insight into spatial memory formation as well as represent tentative pharmacological targets. PMID- 21618642 TI - Histomorphological study of palatal shelf elevation during murine secondary palate formation. AB - During mammalian secondary palate development, the palatal shelves undergo dramatic morphological changes to elevate from a vertical to a horizontal plane in the oral-nasal cavity. We found that E14.5 mouse embryos displayed marked variations in shelf morphology that represent various intermediate states of the elevation process. With these variations, we reconstructed the sequence of shelf morphological changes that take place during the elevation process and discovered distinct patterns in different regions along the anterior-posterior (AP) axis. Moreover, our study revealed that during the elevation process, shelf morphological changes are accompanied by tongue morphological changes, which also show distinct characteristics along the AP axis. We further discuss how to divide the palate along the AP axis based on morphological criteria. Our study provides a framework that recognizes variation in timing of palatal morphogenesis along the AP axis that will aid in the investigation of the mechanisms regulating palatal shelf elevation. PMID- 21618643 TI - The functions of maternal Dishevelled 2 and 3 in the early Xenopus embryo. AB - Of the three Dishevelled (Dvl) genes, only Dvl2 and Dvl3 are maternally encoded in the frog, Xenopus laevis. We show here by loss of function analysis that single depletion of either Dvl2 or Dvl3 from the oocyte causes the same embryonic phenotype. We find that the effects of loss of function of Dvl2 and 3 together are additive, and that the proteins physically interact, suggesting that both are required in the same complex. We show that maternal Dvl2 and 3 are required for convergence extension movements downstream of the dorsally localized signaling pathway activated by Xnr3, but not downstream of the pathway activated by activin. Also, depletion of maternal Dvl2 and 3 mRNAs causes the up-regulation of a subset of zygotic ectodermal genes, including Foxi1e, with surprisingly no significant effect on the canonical Wnt direct target genes Siamois and Xnr3. We suggest that the likely reason for continued expression of the Wnt target genes in Dvl2/3-depleted embryos is that maternal Dvl mRNA depletion is insufficient to deplete stored punctae of Dvl protein in the oocyte cortex, which may transduce dorsal signaling after fertilization. PMID- 21618644 TI - Functional analysis reveals splicing mutations of the CASQ2 gene in patients with CPVT: implication for genetic counselling and clinical management. AB - Catecholaminergic polymorphic ventricular tachycardia (CPVT) is a rare and severe arrhythmogenic disorder. Although usually transmitted in a recessive form, few cases of dominant mutations have been reported. Thirteen mutations in the CASQ2 gene have been reported so far in association with CPVT. We performed molecular analysis of the CASQ2 gene in 43 probands with CPVT and identified eight mutations in five patients. Six mutations were novel: one was a single nucleotide deletion, three affected consensus splice sites, and two had unknown consequences: the c.939 + 5G>C and the synonymous c.381C>T variations. We demonstrated that these two variations affected CASQ2 splicing using a splicing minigene assay. These data increased significantly the number of CASQ2 mutations described in association with CPVT, revealed the high prevalence of splicing and truncating mutations in this gene and brought new insight regarding the dominant inheritance of the disease. Moreover, our report of the first splicing abnormalities in CASQ2 caused by intronic mutation or synonymous change underlines the absolute necessity to perform extensive molecular analysis for genetic diagnosis and counseling of CPVT. PMID- 21618645 TI - A recurrent CHEK2 p.H371Y mutation is associated with breast cancer risk in Chinese women. AB - The association between the CHEK2 and breast cancer risk in Chinese women is unknown. Here, we screened the full CHEK2 coding sequence in 118 Chinese familial breast cancer cases who are negative for mutations in BRCA1 and BRCA2, one recurrent mutation, CHEK2 c.1111C>T (p.H371Y), was identified in five index cases in this cohort. Functional analysis suggested that CHEK2 p.H371Y was a pathogenic mutation that resulted in decreased kinase activity. We further screened the CHEK2 p.H371Y mutation in 909 unselected breast cancer cases and 1,228 healthy individuals. The frequencies of the CHEK2 p.H371Y in familial and unselected breast cancer cases and controls were 4.24% (5/118), 1.76% (16/909), and 0.73% (9/1228), respectively. The p.H371Y mutation was significantly associated with increased breast cancer risk in unselected breast cancer (odds ratio [OR] 2.43, 95% confidence interval [CI] 1.07-5.52, P = 0.034). Our results indicate that the recurrent mutation, p.H371Y, confers a moderate risk of breast cancer in Chinese women. PMID- 21618646 TI - Avoidance of pseudogene interference in the detection of 3' deletions in PMS2. AB - Lynch syndrome is characterized by mutations in the mismatch repair genes MLH1, MSH2, MSH6, and PMS2. In PMS2, detection of mutations is confounded by numerous pseudogenes. Detection of 3' deletions is particularly complicated by the pseudogene PMS2CL, which has strong similarity to PMS2 exons 9 and 11-15, due to extensive gene conversion. A newly designed multiplex ligation-dependent probe amplification (MLPA) kit incorporates probes for variants found in both PMS2 and PMS2CL. This provides detection of deletions, but does not allow localization of deletions to the gene or pseudogene. To address this, we have developed a methodology incorporating reference samples with known copy numbers of variants, and paired MLPA results with sequencing of PMS2 and PMS2CL. We tested a subset of clinically indicated samples for which mutations were either unidentified or not fully characterized using existing methods. We identified eight unrelated patients with deletions encompassing exons 9-15, 11-15, 13-15, 14-15, and 15. By incorporating specific, characterized reference samples and sequencing the gene and pseudogene it is possible to identify deletions in this region of PMS2 and provide clinically relevant results. This methodology represents a significant advance in the diagnosis of patients with Lynch syndrome caused by PMS2 mutations. PMID- 21618647 TI - Analysis of pathway mutation profiles highlights collaboration between cancer associated superpathways. AB - The biological interpretation of the complexity of cancer somatic mutation profiles is a major challenge in current cancer research. It has been suggested that mutations in multiple genes that participate in different pathways are collaborative in conferring growth advantage to tumor cells. Here, we propose a powerful pathway-based approach to study the functional collaboration of gene mutations in carcinogenesis. We successfully identify many pairs of significantly comutated pathways for a large-scale somatic mutation profile of lung adenocarcinoma. We find that the coordinated pathway pairs detected by comutations are also likely to be coaltered by other molecular changes, such as alterations in multifunctional genes in cancer. Then, we cluster comutated pathways into comutated superpathways and show that the derived superpathways also tend to be significantly coaltered by DNA copy number alterations. Our results support the hypothesis that comprehensive cooperation among a few basic functions is required for inducing cancer. The results also suggest biologically plausible models for understanding the heterogeneous mechanisms of cancers. Finally, we suggest an approach to identify candidate cancer genes from the derived comutated pathways. Together, our results provide guidelines to distill the pathway collaboration in carcinogenesis from the complexity of cancer somatic mutation profiles. PMID- 21618648 TI - REEP1 mutations in SPG31: frequency, mutational spectrum, and potential association with mitochondrial morpho-functional dysfunction. AB - Hereditary spastic paraplegias (HSP) constitute a heterogeneous group of neurodegenerative disorders characterized at least by slowly progressive spasticity of the lower limbs. Mutations in REEP1 were recently associated with a pure dominant HSP, SPG31. We sequenced all exons of REEP1 and searched for rearrangements by multiplex ligation-dependent probe amplification (MLPA) in a large panel of 175 unrelated HSP index patients from kindreds with dominant inheritance (AD-HSP), with either pure (n = 102) or complicated (n = 73) forms of the disease, after exclusion of other known HSP genes. We identified 12 different heterozygous mutations, including two exon deletions, associated with either a pure or a complex phenotype. The overall mutation rate in our clinically heterogeneous sample was 4.5% in French families with AD-HSP. The phenotype was restricted to pyramidal signs in the lower limbs in most patients but nine had a complex phenotype associating axonal peripheral neuropathy (= 5/11 patients) including a Silver-like syndrome in one patient, and less frequently cerebellar ataxia, tremor, dementia. Interestingly, we evidenced abnormal mitochondrial network organization in fibroblasts of one patient in addition to defective mitochondrial energy production in both fibroblasts and muscle, but whether these anomalies are directly or indirectly related to the mutations remains uncertain. PMID- 21618649 TI - Dual-color fluorescence lifetime correlation spectroscopy to quantify protein protein interactions in live cell. AB - Dual-color fluorescence correlation spectroscopy is an interesting method to quantify protein interaction in living cells. But, when performing these experiments, one must compensate for a known spectral bleed through artifact that corrupts cross-correlation data. In this article, problems with crosstalk were overcome with an approach based on fluorescence lifetime correlation spectroscopy (FLCS). We show that FLCS applied to dual-color EGFP and mCherry cross correlation allows the determination of protein-protein interactions in living cells without the need of spectral bleed through calibration. The methodology was validated by using EGFP-mCherry tandem in comparison with coexpressed EGFP and mCherry in live cell. The dual-color FLCS experimental procedure where the different laser intensities do not have to be controlled during experiment is really very helpful to study quantitatively protein interactions in live sample. PMID- 21618650 TI - Antennal sensilla of Apanteles cypris Nixon (Hymenoptera: Braconidae), a larval endoparasitoid of Cnaphalocrocis medinalis Guenee (Lepidoptera: Pyralidae). AB - Apanteles cypris is an endoparasitoid of rice pest Cnaphalocrocis medinalis. To better understand its host location mechanism, we examined the external morphology of antennal sensilla of A. cypris via scanning electron microscopy. The antennae of male and female A. cypris were filiform in shape, which consist of scape, pedicel, and flagellum with 18 antennomeres. Eleven types of sensilla were recorded in both sexes, including two types of chaetica sensilla, trichodea sensilla without pore, basiconica sensilla I with a tip pore, basiconica sensilla II with wall pores, basiconica sensilla III with a mastoid at the tip, coeloconica sensilla with apparent fingerlike projections, campaniform sensilla with a sprout tip pore, clavatea sensilla, smell pore, and placodea sensilla (PLA) with the multiporous cuticular structure. We also compared the morphology, number, and distribution of the sensory receptors between the two sexes. The length and width of the antennae on the males were significantly greater than that of females. In addition, males of A. cypris showed a larger number and a higher density of PLA compared to females. The morphology and structure of these sensilla were compared to that found in other braconid wasps, and their possible functions in host location were discussed. These observations could facilitate future studies on the biology of olfaction in A. cypris. PMID- 21618651 TI - Adaptive-weighted cubic B-spline using lookup tables for fast and efficient axial resampling of 3D confocal microscopy images. AB - Confocal laser scanning microscopy has become a most powerful tool to visualize and analyze the dynamic behavior of cellular molecules. Photobleaching of fluorochromes is a major problem with confocal image acquisition that will lead to intensity attenuation. Photobleaching effect can be reduced by optimizing the collection efficiency of the confocal image by fast z-scanning. However, such images suffer from distortions, particularly in the z dimension, which causes disparities in the x, y, and z directions of the voxels with the original image stacks. As a result, reliable segmentation and feature extraction of these images may be difficult or even impossible. Image interpolation is especially needed for the correction of undersampling artifact in the axial plane of three-dimensional images generated by a confocal microscope to obtain cubic voxels. In this work, we present an adaptive cubic B-spline-based interpolation with the aid of lookup tables by deriving adaptive weights based on local gradients for the sampling nodes in the interpolation formulae. Thus, the proposed method enhances the axial resolution of confocal images by improving the accuracy of the interpolated value simultaneously with great reduction in computational cost. Numerical experimental results confirm the effectiveness of the proposed interpolation approach and demonstrate its superiority both in terms of accuracy and speed compared to other interpolation algorithms. PMID- 21618652 TI - Cytotoxicity of fipronil on mice liver cells. AB - In the present study, mice livers were examined following exposure to different doses of fipronil (15, 25, and 50 mg/kg). Histological and histochemical techniques were used to determine the cytotoxic potential of this compound and to assess the damage it caused to livers. Mice were divided into four groups: control group and groups I, II, and III were exposed to 15, 25, and 50 mg/kg fipronil, respectively. Our findings revealed cytological, morphohistological, and histochemical alterations in liver cells of animals from groups I, II, and III compared to group control animals. These changes included Kupffer-cell proliferation, hepatocyte hypertrophy, accumulation and distribution of proteins, polysaccharides, lipids, and vacuoles in the cytoplasm of hepatocytes, and congestion of blood vessels. These phenotypes mainly characterize the following: (a) autophagic processes, (b) steatosis, and (c) cell death by necrosis, which demonstrate the damage caused by fipronil on nontarget organisms in artificial conditions. PMID- 21618653 TI - Effect of alveolex on the bone defects repair stimulated by rhBMP-2: Histomorphometric study. AB - OBJECTIVE: The aim of this study was to evaluate histomorphometrically the effect of alveolex (Propolis 10%) on the repair of bone cavities in the calvaria of rats. MATERIALS AND METHODS: A 5 mm diameter bone defect was made in the calvaria of male Wistar rats using the drill-type trephine. The defects were filled with rhBMP-2+Alveolex, rhBMP-2, Alveolex, or coagulum. Twenty-eight animals with seven subjects on each were sacrificed 30 days after surgery and samples were fixed and embedded in paraffin. Histological sections stained by HE (hematoxylin and eosin) were obtained from the calvaria bone defect and analyzed by a differential point counting method. RESULTS: Group I and II, rhBMP-2+Alveolex and rhBMP-2, respectively, presented higher levels of newly formed bone than other groups (P < 0.001). There were not significant differences between groups I and II (P > 0.05). In addition, there was not significant difference between groups III and IV, Control-Coagulum and Alveolex, respectively (P > 0.05). CONCLUSION: Alveolex has increased the bone repair in calvaria defects of rats when associated to rhBMP-2, however without significant differences for rhBMP-2 isolated group; Alveolex isolated group showed the lowest levels of newly formed bone with no significant differences to coagulum group (control). PMID- 21618654 TI - In vitro evaluation of the microbial contamination on new toothbrushes: a preliminary study. AB - OBJECTIVE: The presence and survival of microorganisms on toothbrush bristles might play a role on the etiology of oral infections. The aim of this in vitro study was to evaluate the presence of bacterial contamination on new toothbrushes before oral contact. MATERIALS AND METHODS: Forty toothbrushes from five different manufacturers were used in this experimental study. Each manufacturer was divided according to conventional local of obtaining: industry, drugstore, market, and perfumery. The toothbrush heads were completely immersed into tubes containing 5.0 mL of sterile peptonated water (dilution 1:10). A group of eight tubes containing the sterile solution was used as control. After 21 days of anaerobic incubation, occurrence of contamination was visually evaluated and confirmed by light microscopy. RESULTS: Bacterial growth in the medium, indicative of bristles contamination, was found in a total of 19 out of 40 samples (47.5%) evaluated: 6 out of 14 samples (42.85%) from industry group, 4 out of 8 samples (50.0%) from drugstore, 5 out of 10 samples (50.0%) from market, and 4 out of 8 samples (50.0%) from perfumery. Only the toothbrushes with bristles coated with chlorhexidine did not show contamination. The Gram-negative sporulating bacilli were the most prevalent form recovered. CONCLUSIONS: Except for chlorhexidine group, bacterial growth was observed in all groups evaluated irrespective local of obtaining. PMID- 21618655 TI - Morphological alterations in the prostate stroma of rats submitted to chronic nicotine treatment. AB - The stroma plays a fundamental role in the function of different glandular systems. In the prostate, the stroma is responsible for the development and maintenance of the differentiated state of the epithelium. Nicotine induces tobacco dependence and promotes morphological alterations in the epithelial compartment. However, its effects on the prostate stroma are unclear. The objective of this study was to evaluate the morphology of the stromal microenvironment in the ventral prostate lobe of rats submitted to chronic nicotine administration. Twenty rats (Rattus norvegicus) were divided into two groups: 10 animals received subcutaneous nicotine and 10 animals received physiological saline by the same route. After treatment, samples were collected from the ventral prostate lobe, processed and submitted to histology, histochemistry, and ultrastructural analysis by transmission and scanning electron microscopy. The level of circulating testosterone was also analyzed. The results showed a significant increase in the density of type I collagen (56.3% to 85.9%, P < 0.01) and a decrease in the density of type III collagen (43.7% to 14.1%, P < 0.01). In addition, there was a qualitative increase in elastic fibers and in the number of smooth muscle cells with a secretory phenotype. Circulating testosterone levels were significantly reduced (898.3 to 363.1 ng/mL, P < 0.01). The results showed that nicotine modifies different components of the prostate stroma, suggesting that this drug is a risk factor for morphofunctional alterations in the prostate gland. PMID- 21618656 TI - Application of microscopy in authentication of valuable Chinese medicine I- Cordyceps sinensis, its counterfeits, and related products. AB - Light and polarized microscope was applied to authenticate 32 Cordyceps and 6 artificial counterfeits and 8 fermented Cordyceps as well as 7 Cordyceps capsules available in Hong Kong markets. Results showed that transverse sections of stroma and powder of larvae can be used to differentiate C. sinensis from its counterfeits. The fermented Cordyceps are in powder form. Among the eight fermented Cordyceps collected, half of them were pure; three were a mixture of fermented Cordyceps and soya beans; one was a mixture of unknown plant tissues and soya beans. For the seven Cordyceps capsules, the powders of five samples were a mixture of fermented Cordyceps and soya beans; the powders of other two were a mixture of C. sinensis stroma powder and fermented Cordyceps. The study indicated that the microscopy is an unambiguous method that requests fewer sample for the authentication of valuable Chinese medicine-C. sinensis and its related products. PMID- 21618657 TI - An imaging technique using rotational polarization microscopy for the microstructure analysis of carbon/carbon composites. AB - A novel image analysis technique was proposed for microstructure investigation of carbon/carbon (C/C) composites. The rotational polarization microscopy was developed to meet the special imaging requirements. The samples of C/C composites were observed in reflection polarized light microscope, where the analyzer was rotated instead of the stage, and the polarizer was taken out. The bireflectance of like-graphite negative uniaxial crystal was analyzed. It was the theoretic foundation of image collection and data processing. The analyzer was rotated through 36 * 10 degrees intervals without any movement of the specimen. The polished cross-section of C/C composites took micrographs at each analyzer orientation. All image data collected from the same field of view were processed by image registration and image fusion. The synthesized images were obtained by calculating the maximum and minimum gray values and their differences at each point of the million pixels at 18 orientations of the analyzer. They are unique and quite reliable to be applied to analyze the microstructure of C/C composites. Subsequently, image segmentation was performed, and the feature parameters of each component were calculated. Good agreement was found between the results from image analysis and experimental data. PMID- 21618658 TI - White matter integrity and behavioral activation in healthy subjects. AB - Individual differences in behavioral inhibition and behavioral activation may place certain people at greater risk for neuropsychiatric disorders and engagement in risky behaviors. Therefore, studying the neural correlates of behavioral inhibition and activation may help us understand neural mechanisms underlying risk behaviors in both clinical and non-clinical populations. To investigate, we assessed the relationships between white matter integrity and measures of behavioral inhibition and behavioral activation in 51 healthy participants using diffusion tensor imaging (DTI) and the Behavioral Inhibition System/Behavioral Activation System (BIS/BAS) scale. Scores on the Fun-Seeking subscale of the BAS positively correlated with DTI fractional anisotropy in the left corona radiata and adjacent superior longitudinal fasciculus, and with mean diffusivity in the left inferior longitudinal fasciculus and inferior fronto occipital fasciculus after controlling for age, gender, and education. These findings suggest that the integrity of white matter connecting extensive brain regions implicated in self-control and the processing of rewards and emotions are associated with individual differences in the motivation for seeking and participating in fun and novel experiences. PMID- 21618659 TI - sLORETA allows reliable distributed source reconstruction based on subdural strip and grid recordings. AB - Source localization based on invasive recordings by subdural strip and grid electrodes is a topic of increasing interest. This simulation study addresses the question, which factors are relevant for reliable source reconstruction based on sLORETA. MRI and electrode positions of a patient undergoing invasive presurgical epilepsy diagnostics were the basis of sLORETA simulations. A boundary element head model derived from the MRI was used for the simulation of electrical potentials and source reconstruction. Focal dipolar sources distributed on a regular three-dimensional lattice and spatiotemporal distributed patches served as input for simulation. In addition to the distance between original and reconstructed source maxima, the activation volume of the reconstruction and the correlation of time courses between the original and reconstructed sources were investigated. Simulations were supplemented by the localization of the patient's spike activity. For noise-free simulated data, sLORETA achieved results with zero localization error. Added noise diminished the percentage of reliable source localizations with a localization error <=15 mm to 67.8%. Only for source positions close to the electrode contacts the activation volume correctly represented focal generators. Time-courses of original and reconstructed sources were significantly correlated. The case study results showed accurate localization. sLORETA is a distributed source model, which can be applied for reliable grid and strip based source localization. For distant source positions, overestimation of the extent of the generator has to be taken into account. sLORETA-based source reconstruction has the potential to improve the localization of distributed generators in presurgical epilepsy diagnostics and cognitive neuroscience. PMID- 21618660 TI - Neural basis of the association between depressive symptoms and memory deficits in nondemented subjects: resting-state fMRI study. AB - Depressive symptoms often coexist with memory deficits in older adults and also are associated with incident cognitive decline in the elderly. However, little is known about the neural correlates of the association between depressive symptoms and memory deficits in nondemented elderly. Fifteen amnestic mild cognitive impairment (aMCI) and 20 cognitively normal (CN) subjects completed resting-state functional magnetic resonance imaging (R-fMRI) scans. Multiple linear regression analysis was performed to test the main effects of the Geriatric Depression Scale (GDS) and Rey Auditory Verbal Learning Test delayed recall (RAVLT-DR) scores, and their interaction on the intrinsic amygdala functional connectivity (AFC) network activity. Severer depressive symptoms and memory deficits were found in the aMCI group than in the CN group. Partial correlation analysis identified that the RAVLT-DR scores were significantly correlated with the AFC network in the bilateral dorsolateral prefrontal cortex (DLPFC), dorsomedial and anterior prefrontal cortex, posterior cingulate cortex (PCC), middle occipital gyrus, right inferior parietal cortex, and left middle temporal gyrus (MTG). The GDS scores were positively correlated with the AFC network in the bilateral PCC and MTG, and left DLPFC. The interactive effects of the GDS and RAVLT-DR scores on the AFC network were seen in the bilateral PCC, MTG, and left DLPFC. These findings not only supported that there were interactive neural links between depressive symptoms and memory functions in nondemented elderly at the system level, but also demonstrated that R-fMRI has advantages in investigating the interactive nature of different neural networks involved in complex functions, such as emotion and cognition. PMID- 21618661 TI - Overnight deprivation from smoking disrupts amygdala responses to fear. AB - Cigarette smoking, a major, yet avoidable, cause of disability and premature death, is the most prevalent form of nicotine addiction. An emerging theme in the neurobiology of nicotine addiction is the integrity of the amygdala. Using functional MRI, amygdala responses during a face perception task were compared between 28 chronic smokers [14 females, 14 males; age, 26.3 (2.8) years; age at onset of smoking, 15.8 (2.6) years; years smoked, 9.1 (2.1); cigarettes per day, 17.1 (3.7); Fagerstrom test for nicotine dependence score, 4.1 (1.9); exhaled carbon-monoxide level, 17.8 (9.5) ppm] and 28 age- and education-matched nonsmokers [14 females, 14 males; age, 26.9 (2.4) years]. Subjects underwent imaging on two separate occasions 1 week apart: smoking satiety versus overnight smoking deprivation, in a randomized counterbalanced order. Our results show no difference in amygdala responses to faces between nonsmokers and satiated smokers. However, overnight deprivation from smoking was associated with a significantly lowered amygdala response to fear, an effect that was probabilistically mapped to the basolateral amygdala. We suggest that aberrant amygdala reactivity in overnight-deprived smokers may reflect a pre-existing vulnerability to smoking and/or increase the risk of smoking relapse after a cessation attempt. PMID- 21618662 TI - Quantifying change in individual subjects affected by frontotemporal lobar degeneration using automated longitudinal MRI volumetry. AB - A novel method of automated MRI volumetry was used to study regional atrophy and disease progression in repeated MRI measurements of patients with frontotemporal lobar degeneration (FTLD). Fifty-nine structural MRI data sets of 17 clinically diagnosed FTLD patients were acquired over up to 30 months in intervals of 6 months and compared with data of 30 age-matched healthy controls. Patients were further subgrouped into behavioral variant FTLD (bvFTLD), progressive nonfluent aphasia (PNFA), and semantic dementia (SemD). Gray matter (GM) volumes of frontal lobes (FL) and temporal lobes (TL) were determined by voxel-based volumetry based on SPM5 algorithms and a probabilistic brain atlas. MRI volumetry revealed frontal and temporal GM atrophy across FTLD patients, with further progression over time. Significant side asymmetry of TL volumes was found in SemD. The ratio of TL to FL volumes was significantly reduced in SemD and increased in bvFTLD. Using this ratio, 6/7 SemD patients and 5/6 bvFTLD patients could be correctly differentiated. TL/FL ratios in bvFTLD and SemD further diverged significantly over a time span of only 6 months. Rates of temporal GM loss per 6 months were 3 4% in SemD, and 2.5% for frontal GM loss in bvFTLD, and thereby clearly exceeded published cerebral volume loss in healthy elderly subjects. The study presents a fully automated, observer-independent volumetric assessment of regional atrophy which allows differentiation of FTLD subgroups. Its sensitivity for atrophy progression--even in such short intervals like 6 months--might benefit future clinical trials as treatment outcome measure. PMID- 21618663 TI - Coupling electrophysiological and hemodynamic responses to errors. AB - Performance errors are associated with distinct electrophysiological and hemodynamic signatures: a fronto-central error-related negativity (ERN) is seen in the event-related potentials and a network of activations including medio frontal, parietal, and insular cortex is revealed by functional magnetic resonance imaging. We used simultaneous electroencephalography and functional magnetic resonance imaging (fMRI) to characterize the relationship between the electrophysiological and hemodynamic responses to errors. Participants performed a modified Flanker task. When analyzed independently, we found the ERN and hemodynamic activations in dorsal anterior cingulate cortex, superior frontal gyrus, precentral gyrus, inferior frontal gyrus, and inferior parietal lobule. fMRI-informed dipole modeling and joint independent component analysis (ICA) were used to couple electrophysiological and hemodynamic data. Both techniques revealed a temporal evolution of the areas found in the fMRI analysis, with the right hemisphere activations peaking before the left hemisphere. However, joint ICA added information, revealing a number of cortical and subcortical areas that had not been shown with parametric mapping. This technique also uncovered how these areas evolve over time. All together, these analyses provide a more detailed picture of the spatiotemporal dynamics of the processing of performance errors. PMID- 21618664 TI - fMRI investigation of speed-accuracy strategy switching. AB - Switching between rapid and accurate responses is an important aspect of decision making. However, the brain mechanisms important to smoothly change the speed accuracy strategy remain mostly unclear. This issue was addressed here by using functional magnetic resonance imaging (fMRI). On each trial, right-handed healthy participants had to stress speed or accuracy in performing a color discrimination task on a target stimulus according to the instructions given by an initial cue. Participants were capable of trading speed for accuracy and vice versa. Analyses of cue-related fMRI activations revealed a significant recruitment of left middle frontal gyrus and right cerebellum when switching from speed to accuracy. The left superior parietal lobule was activated in the same switching condition but only after the target onset. The anterior cingulate cortex was more recruited, also after target presentation, when speed had to be maintained from one trial to the next. These results are interpreted within a theoretical framework that attributes a role in criterion-setting to the left lateral prefrontal cortex, perceptual evidence accumulation to the superior parietal lobule, and action energization to the anterior cingulate cortex, extending previous findings to the domain of speed-accuracy tradeoff regulations. PMID- 21618665 TI - Oscillatory correlates of controlled speed-accuracy tradeoff in a response conflict task. AB - In making decisions, people have to balance between the competing demands of speed and accuracy, a balance generally referred to as the speed-accuracy tradeoff (SAT). In this study, we investigated the role of controlled SAT in a two-choice task in which manual responses were either validly or invalidly cued. Examining electrophysiological measurements of oscillatory brain activity, theta power in the anterior cingulate cortex (ACC), alpha power in the occipital cortex, and beta power in the motor cortex were found to be related to SAT. Because oscillatory effects of SAT were found to emanate from the SAT baseline interval preceding the two-choice task, the results indicate that SAT is modulated by a change of visuo-motor baseline activities rather than a change of response threshold. Moreover, in the two-choice task, conflict-induced theta power in the ACC was found to be more pronounced in speed than in accuracy trials, whereas priming-related beta power dynamics in the motor cortex were unaffected by SAT. These results indicate that conflict processing, but not response priming, depends on SAT. PMID- 21618666 TI - Brain activation for language dual-tasking: listening to two people speak at the same time and a change in network timing. AB - The study used fMRI to investigate brain activation in participants who were able to listen to and successfully comprehend two people speaking at the same time (dual-tasking). The study identified brain mechanisms associated with high-level, concurrent dual-tasking, as compared with comprehending a single message. Results showed an increase in the functional connectivity among areas of the language network in the dual task. The increase in synchronization of brain activation for dual-tasking was brought about primarily by a change in the timing of left inferior frontal gyrus (LIFG) activation relative to posterior temporal activation, bringing the LIFG activation into closer correspondence with temporal activation. The results show that the change in LIFG timing was greater in participants with lower working memory capacity, and that recruitment of additional activation in the dual-task occurred only in the areas adjacent to the language network that was activated in the single task. The shift in LIFG activation may be a brain marker of how the brain adapts to high-level dual tasking. PMID- 21618667 TI - Racial identification modulates default network activity for same and other races. AB - Racial identification shapes self-concept and how people share in and respond to the emotional states of others around them. Prior neuroimaging studies have demonstrated the role of the neural default network in self-referential and empathic processing. However, how racial identification affects neural processing of social information remains unknown. Here, we examined the effect of racial identification on neural response related to social perception among African American and Caucasian American individuals using functional magnetic resonance imaging. Our results demonstrate that degree of racial identification predicts activity within cortical midline structures of the default network in response to viewing racial ingroup, relative to outgroup members, and activity within the medial temporal lobe subsystem of the default network in response to viewing racial outgroup, relative to ingroup members. Broadly, our findings suggest that the strength of racial identification is associated with differential recruitment of neural and cognitive processes to understand and respond to other people within and outside of one's racial group. PMID- 21618668 TI - Comprehension of self-report evidence-based measures of anxiety. AB - BACKGROUND: Given their applicability in diverse settings and for a wide range of purposes, the generalizability of self-report symptom measures is particularly important. An understudied factor in the development and validation of self report measures is the degree to which they are difficult to comprehend. This study evaluated the difficulty of self-report measures of anxiety with respect to several domains, including formatting, length, and linguistic problems. METHODS: Ninety-two evidence based measures of anxiety were evaluated for comprehension level. RESULTS: The majority of anxiety measures included challenging elements of formatting, linguistic ability, and readability. Measures of obsessive-compulsive disorder were associated with the highest level of comprehension (i.e., greatest difficulty). CONCLUSIONS: The validity of self-report measures relies on the ability of respondents to understand the instructions and measure items. Factors related to the comprehension of self-report measures should be included among the basic psychometric properties in measure development and validation. Future research on the development of self-report measures that can be more broadly applicable across levels of education and literacy are of particular importance to research, clinical, and public health agendas. PMID- 21618669 TI - What little we know about tailoring depression treatment for individual patients. PMID- 21618670 TI - Generalized anxiety disorder and anorexia nervosa: evidence of shared genetic variation. AB - BACKGROUND: Previous studies have indicated a high prevalence of generalized anxiety disorder (GAD) in women with anorexia nervosa (AN). However, the shared genetic and environmental components of these disorders have not been explored. This study seeks to elucidate the shared genetic and environmental components between GAD and AN. METHOD: Using 2,083 women from the Virginia Adult Twin Study of Psychiatric and Substance Use Disorders, structural equation modeling was used to obtain maximum likelihood estimates of the environmental genetic, shared and unique environmental components in 496 women with GAD, 47 women with AN, 43 women with GAD + AN, and 1,497 women without GAD or AN. RESULTS: Results show that the heritability of GAD was 0.32 and AN was 0.31, and the genetic correlation between the two disorders was 0.20, indicating a modest genetic contribution to their comorbidity. Unique environment estimate was 0.68 for GAD and 0.69 for AN, with a unique environmental correlation of 0.18. All common environmental parameters were estimated at zero. CONCLUSIONS: The modest shared genetic and unique environmental liability to both disorders may help explain the high prevalence of GAD in women with AN. This knowledge could help in the treatment and prevention of comorbid disorders. PMID- 21618671 TI - Olfactory identification and discrimination in obsessive-compulsive disorder. AB - BACKGROUND: Neuroimaging and neuropsychological data from patients with an obsessive-compulsive disorder (OCD) indicate the dysfunction of the orbitofrontal cortex (OFC). Olfactory processing has been associated with OFC function, although results from OCD studies regarding this sensory modality have been inconclusive. No previous study has analyzed both odor discrimination and identification capacity in OCD patients using "Sniffin' Sticks" tests. The aim of our study was to assess odor threshold, identification, discrimination, and nonverbal memory in OCD patients, in order to determine whether these functions were affected. METHODS: Olfactory function was measured in 29 OCD patients and 29 healthy volunteers (HV) using the "Sniffin' Sticks" test and their nonverbal memory was scored with the Rey-Osterrieth Complex Figure Test. RESULTS: OCD patients showed significant impairment in their odor performance and in their execution of the nonverbal memory task compared to HV. No statistical associations were found between the deficits in the two areas. The severity of depressive and obsessive-compulsive symptoms did correlate with olfactory identification. CONCLUSION: Our findings support the hypothesis that olfactory and memory dysfunctions in OCD reflect different neurobiological alterations of the disorder, and point to the modulation effect of depressive and obsessive compulsive symptoms on odor performance. PMID- 21618672 TI - Differences in the determinants of posttraumatic stress disorder and depression after a mass traumatic event. AB - BACKGROUND: Hurricane Ike struck the Galveston Bay area of Texas on September 13, 2008, leaving substantial destruction and a number of deaths in its wake. We assessed differences in the determinants of posttraumatic stress disorder (PTSD) and depression after this event, including the particular hurricane experiences, including postevent nontraumatic stressors, that were associated with these pathologies. METHODS: 658 adults who had been living in Galveston and Chambers counties, TX in the month before Hurricane Ike were interviewed 2-5 months after the hurricane. We collected information on experiences during and after Hurricane Ike, PTSD and depressive symptoms in the month before the interview, and socio demographic characteristics. RESULTS: The prevalence of past month hurricane related PTSD and depression was 6.1 and 4.9%, respectively. Hurricane experiences, but not socio-demographic characteristics, were associated with Ike related PTSD. By contrast, lower education and household income, and more lifetime stressors were associated with depression, as were hurricane exposures and hurricane-related stressors. When looking at specific hurricane-related stressors, loss or damage of sentimental possessions was associated with both PTSD and depression; however, health problems related to Ike were associated only with PTSD, whereas financial loss as a result of the hurricane was associated only with depression. CONCLUSIONS: PTSD is indeed a disorder of event exposure, whereas risk of depression is more clearly driven by personal vulnerability and exposure to stressors. The role of nontraumatic stressors in shaping risk of both pathologies suggests that alleviating stressors after disasters has clear potential to mitigate the psychological sequelae of these events. PMID- 21618673 TI - Synthesis of 2-keto-anti-1,3-diols by chemoselective tandem oxidation of 2-B(pin) substituted allylic alcohols. PMID- 21618674 TI - Main-chain heterobimetallic block copolymers: synthesis and self-assembly of polyferrocenylsilane-b-poly(cobaltoceniumethylene). PMID- 21618675 TI - Identification of novel quadruplex ligands from small molecule libraries by FRET based high-throughput screening. PMID- 21618676 TI - Bio-distribution and in vivo antioxidant effects of cerium oxide nanoparticles in mice. AB - Cerium oxide nanoparticles have oxygen defects in their lattice structure that enables them to act as a regenerative free radical scavenger in a physiological environment. We performed a comprehensive in vivo analysis of the biological distribution and antioxidant capabilities of nanoceria administered to mice perorally (PO), intravenously (IV), or intraperitoneally (IP) by dosing animals weekly for 2 or 5 weeks with 0.5 mg kg(-1) nanoceria. Next, we examined if nanoceria administration would decrease ROS production in mice treated with carbon tetrachloride (CCl(4)). Our results showed that the most extensive and cumulative nano-deposition was via IV and IP administered while PO administration showed mice excreted greater than 95% of their nanoceria within 24 h. Organ deposition for IV and IP mice was greatest in the spleen followed by the liver, lungs, and kidneys. Elimination for all administration routes was through feces. Nanoceria administration showed no overt toxicity, however, WBC counts were elevated with IV and IP administration. Our in vivo studies show that nanoceria administration to mice with induced liver toxicity (by CCl(4)) showed similar findings to mice treated with N-acetyl cystine (NAC), a common therapeutic to reduce oxidative stress. Taken together, our studies show that nanoceria remains deposited in tissues and may decrease ROS, thereby suggesting that cerium oxide nanoparticles may be a useful antioxidant treatment for oxidative stress. PMID- 21618677 TI - The toxic effects of nickel chloride on liver, erythropoiesis, and development in Wistar albino preimplanted rats can be reversed with selenium pretreatment. AB - The exposure to nickel chloride (NiCl2) can cause hematotoxicity and hepatotoxicity and canaffect development. The present study pertains to the protective effect of selenium (Se) against NiCl2-induced toxicity in preimplanted Wistar albino rats. The subcutaneous (s.c.) administration of 25 or 50 mg/kg of NiCl2 to Wistar albino rats on day 3 of gestation induced an immediate and significant decrease in maternal body weight and anemia 2 days after treatment. In addition, an increase in plasma aspartate aminotransferase (AST) was observed. These effects were maintained on day 20 of gestation. Moreover, a significant increase in plasma alanine aminotransferase (ALT) levels was observed with the administration of 25 mg/kg of NiCl2. Conversely, administration of 50 mg/kg of NiCl2 by s.c. injection increased erythropoiesis at day 20 of gestation and decreased platelets counts. In addition, administration of 100 mg/kg of NiCl2 markedly reduced the maternal body weight and number of live fetuses and increased fetal loss, predominantly at the end of the experimental period. All dose levels of NiCl2 caused an alteration in the hepatic histoarchitecture. When 0.3-mg/kg Se was injected s.c. with 100-mg/kg NiCl2, the levels of plasma AST and ALT and the structure of the liver were restored. Administration of 20 mg/L/day of NiCl2 in the drinking water significantly reduced the maternal body weight at day five of gestation as well as erythropoiesis during the exposure period. The present study suggests that Se can counteract the nocuous effect of nickel on the liver; however this antioxidant did not prevent alterations in development and erythropoiesis. PMID- 21618678 TI - Impact of anticoagulation on ionic and nonionic contrast media effect on thrombogenesis and fibrinolysis: The PEPCIT study. AB - OBJECTIVES: The effect of ionic low osmolar contrast media (ICM) and nonionic iso osmolar CM (NICM) on acute thrombotic complications of percutaneous coronary intervention (PCI) is subject to controversies possibly related to a potential interaction with anticoagulation regimens. We sought to compare physical and morphological properties of fibrin clots made in the presence of ioxaglate (ICM), iodixanol (NICM) versus control and to evaluate the effect of four anticoagulants used in PCI. METHODS AND RESULTS: Maximum platelet aggregation (MPA%), maximum elastic modulus (EM, dyne/cm(2) ) fiber density (n/10(-5) /MUm(2) ), and lysis front velocity (nm/sec) of fibrin rich clot (FRC) were measured simultaneously using peripheral blood from 12 patients undergoing elective PCI. We compared the effects of adding iodixanol or ioxaglate or saline (control) to blood with enoxaparin, unfractionated heparin, fondaparinux, and bivalirudin. Iodixanol and ioxaglate led to nonsignificant reduction in MPA compared to control (33.6% +/- 16.9%, 28.2% +/- 18.9%, and 40.7% +/- 13.9%, respectively, P = ns). Fibrin formed with iodixanol was stiffer (42.7 +/- 41.9, 18.7 +/- 3.7, and 15.9 +/- 9 dyne/cm(2) , P < 0.01) and displayed more fibrin fibers (1089 +/- 175, 260 +/- 108, and 456 +/- 131 n/10(-5) /MUm(2) , respectively, P < 0.01) than with ioxaglate or control. This resulted in a profound reduction in the lysis front velocity (191 +/- 95, 261 +/- 112, and 360 +/- 153 nm/sec). None of the four anticoagulants displayed any significant interaction on the effect of contrast media. CONCLUSIONS: The prothrombogenic effect of iodixanol is related primarily to an increase in fibrin stiffness with subsequent delayed fibrinolysis, something not seen with ioxaglate. Anticoagulation does not appear to have any impact on this fibrin clot abnormalities. PMID- 21618679 TI - Optimizing antiplatelet therapy in acute coronary syndrome and percutaneous coronary intervention. AB - Dual antiplatelet therapy with aspirin and clopidogrel is the standard of care for patients with acute coronary syndrome (ACS) and those undergoing percutaneous coronary intervention (PCI). It is well established that inhibition of platelet aggregation reduces the risk of recurrent thrombotic events and stent thrombosis. However, some patients show a reduced antiplatelet response to standard clopidogrel loading (300 mg) and maintenance (75 mg day(-1)) doses, which has been associated with poorer patient outcomes. Pharmacodynamic and pharmacokinetic studies show that higher-than-standard clopidogrel dosing strategies facilitate more rapid platelet inhibition of a greater intensity as a result of greater plasma concentrations of the clopidogrel active metabolite. Recently completed studies suggest that in patients with ACS undergoing PCI, higher-than-standard clopidogrel dosing regimens provide greater inhibition of platelet function and improved clinical outcomes with a small but significant increase in major bleeding. Newer, more potent antiplatelet agents such as prasugrel and ticagrelor are other alternative strategies that result in more rapid, greater inhibition of platelet function and better outcomes than standard-dose clopidogrel. Whether platelet reactivity-guided therapy or genotyping for cytochrome P450 polymorphisms is useful in managing patients needs to be further defined. Most importantly, early and effective antiplatelet therapy results in the best short- and long-term outcomes for patients with ACS or those undergoing PCI. PMID- 21618680 TI - Quantum efficiencies, absolute intensities and signal-to-blackbody ratios of high temperature laser-induced thermographic phosphors. AB - There are a number of issues related to high-temperature phosphor thermometry, which include measurement of faster decays, decreasing emission intensity and rising levels of blackbody radiation, that will impose limits on the maximum delectable temperature. This paper provides absolute intensity measurements, quantum efficiencies and signal-to-blackbody radiation ratios at peak emission wavelengths, at various temperatures (20-1400 degrees C), for Y(2)O(3):Eu, YAG:Tb and YAG:Tm thermographic phosphors under 266 and 355 nm excitation from a Q switched Nd:YAG laser. These terms are beneficial in a number of ways for engineers wanting to use a phosphor thermometry solution at high temperatures. They may also provide additional insight to the physical luminescence processes of phosphors at high temperatures. The phosphor signal:blackbody radiation ratio is useful because it combines the effects of blackbody radiation and phosphor emission intensities at various temperatures, providing a valuable quantitative evaluation that can be used as a design aid for phosphor selection. A figure of merit is the temperature when the blackbody radiation equals the phosphor emission (ratio = 1); this is the cross-over temperature at which the blackbody radiation rapidly starts to overtake and mask out phosphor emissions. To the best of our knowledge no such work exists previously. The results presented show a variation in phosphor intensity with increasing temperature, and although the intensity and quantum efficiencies for Y(2)O(3):Eu and YAG:Tb were much greater than YAG:Tm at low temperatures, YAG:Tm was found to be the most efficient phosphor investigated at higher temperatures (>900 degrees C). With a peak emission wavelength of 458 nm, YAG:Tm experienced the lowest proportion of blackbody radiation therefore its advantage at higher temperatures was further amplified and was found to offer an advantage of approximately +350 degrees C and +250 degrees C increased upper temperature capability compared to Y(2)O(3):Eu and YAG:Tb phosphors, respectively. PMID- 21618681 TI - Dy3+-activated NaM4(VO4)3 (M = Ca, Ba, Sr) phosphor for near-UV solid-state lighting. AB - We report the photoluminescence characterization of Dy(3+)-activated NaM(4)(VO(4))(3) (M = Ca, Ba, Sr) phosphors prepared by a solid-state method. The synthesis was confirmed by X-ray diffraction (XRD) characterization and photoluminescence (PL) emission results showed sharp blue and yellow bands for NaM(4)(VO(4))(3) (M = Ca, Ba, Sr):Dy(3+) phosphors at the excitation wavelength of 323 nm, which is near-UV excitation. Thus, these phosphors could be applicable for near-UV excited solid-state lighting devices. PMID- 21618682 TI - Luminescence properties of a nanoporous freshwater diatom. AB - Freshwater diatom frustules show special optical properties. In this paper we observed luminescence properties of the freshwater diatom Cyclotella meneghiniana. To confirm the morphological properties we present scanning electron microscopy (SEM) images. X-ray diffraction (XRD) studies were carried out to visualize the structural properties of the frustules, confirming that silica present in diatom frustules crystallizes in an alpha-quartz structure. Study of the optical properties of the silica frustules of diatoms using ultra violet-visible (UV-vis) spectroscopy and photoluminescence spectroscopy confirmed that the diatom C. meneghiniana shows luminescence in the blue region of the electromagnetic spectrum when irradiated with UV light. This property of diatoms can be exploited to obtain many applications in day-to-day life. Also, using time resolved photoluminescence spectroscopy (TRPL) it was confirmed that this species of diatom shows bi-exponential decay. PMID- 21618683 TI - The price of "doing the right thing". PMID- 21618684 TI - Use of liver grafts from donation after cardiac death donors for recipients with hepatitis C virus. AB - Hepatitis C virus (HCV) infection is the most common indication for orthotopic liver transplantation in the United States. Although studies have addressed the use of expanded criteria donor organs in HCV(+) patients, to date the use of liver grafts from donation after cardiac death (DCD) donors in HCV(+) patients has been addressed by only a limited number of studies. This retrospective analysis was undertaken to study the outcomes of DCD liver grafts used in HCV(+) recipients. Seventy-seven HCV(+) patients who received DCD liver grafts were compared to 77 matched HCV(+) patients who received donation after brain death (DBD) liver grafts and 77 unmatched non-HCV patients who received DCD liver grafts. There were no differences in 1-, 3-, and 5-year patient or graft survival among the groups. Multivariate analysis showed that the Model for End-Stage Liver Disease score [hazard ratio (HR) = 1.037, 95% confidence interval (CI) = 1.006 1.069, P = 0.018] and posttransplant cytomegalovirus infection (HR = 3.367, 95% CI = 1.493-7.593, P = 0.003) were significant factors for graft loss. A comparison of the HCV(+) groups for fibrosis progression based on protocol biopsy samples up to 5 years post-transplant did not show any difference; in multivariate analysis, HCV genotype 1 was the only factor that affected progression to stage 2 fibrosis (genotype 1 versus non-1 genotypes: HR = 2.739, 95% CI = 1.047-7.143, P = 0.040). In conclusion, this match-controlled, retrospective analysis demonstrates that DCD liver graft utilization does not cause untoward effects on disease progression or patient and graft survival in comparison with DBD liver grafts in HCV(+) patients. PMID- 21618685 TI - Assessment of a chloride-poor versus a chloride-containing version of a modified histidine-tryptophan-ketoglutarate solution in a rat liver transplantation model. AB - Recent in vitro studies of cold-induced cell injury have revealed the detrimental effects of extracellular chloride on cold-stored isolated rat hepatocytes; however, its influence on endothelial cells is beneficial. To determine which of these effects is predominant in vivo, we tested both a chloride-poor variant of a new histidine-tryptophan-ketoglutarate (HTK)-based preservation solution and a chloride-containing variant in a rat liver transplantation model. The study, which was carried out in a blinded fashion with 7 or 8 rats per group, was divided into 2 parts: (1) a comparison of survival in 3 series under different conditions [different microsurgeons, rat strains, cold ischemia times (3, 12, and 24 hours), and warm ischemia times] and (2) an assessment of the microcirculation (30-90 minutes after reperfusion), laboratory data, bile production, and histology. In each of the survival experiments, a (strong) tendency toward prolonged survival was observed with the new chloride-containing solution (50% versus 12.5%, 75% versus 37.5%, and 100% versus 71.4% [chloride-containing vs. chloride-poor], overall P < 0.05). Additionally, the sinusoidal perfusion rates (83.9% +/- 4.0% versus 69.2% +/- 10.8%, P < 0.01) and the red blood cell velocities in sinusoids (147.7 +/- 26.7 versus 115.5 +/- 26.0 MUm/second, P < 0.05) and in postsinusoidal venules (332.4 +/- 87.3 versus 205.5 +/- 53.5 MUm/second, P < 0.01) were clearly higher with chloride. Moreover, the serum activities of liver enzymes were slightly reduced (not significantly), and bile production was significantly increased. These results suggest an overall beneficial effect of chloride in HTK-based liver preservation solutions. PMID- 21618686 TI - Liver transplantation for propionic acidemia in children. AB - Propionic acidemia (PA) is a rare inherited disorder of branched chain amino acid metabolism; despite improvements in conventional medical management, the long term outcome remains disappointing. Liver transplantation (LT) has been proposed to minimize the risk of further metabolic decompensations and to improve the quality of life. We performed a retrospective review of all children with PA who underwent LT between 1987 and 2008. Five children were identified with a median age of 1.2 years (range = 0.7-4.1 years) at referral. Four of the children presented clinically at 3 weeks of age or less, and 1 child was diagnosed prenatally. All had metabolic acidosis and hyperammonemia. Two had seizures and required intensive care; this care included inotropic support and continuous venovenous hemofiltration in 1 child. The children were considered for elective LT for the following reasons: frequent metabolic decompensations (2), previous sibling death (2), and elective management (1). One child underwent auxiliary LT, and 4 children received orthotopic grafts (1 living related graft). The median age at LT was 1.5 years (range = 0.8-7.0 years). There was 1 retransplant 3 months after LT due to hepatic artery thrombosis. One year after LT, 1 patient suffered a metabolic stroke with minimal residual neurology. After a median follow-up of 7.3 years (range = 2.2-15.0 years), all the children had normal graft function and a good quality of life with a protein-unrestricted diet and no further metabolic decompensations. In conclusion, LT has a role in the management of PA: it reduces the risk of metabolic decompensation and improves the quality of life. The potential for the development of metabolic sequelae is not completely eliminated. PMID- 21618687 TI - Splenic artery embolization for the treatment of refractory ascites after liver transplantation. AB - Refractory ascites (RA) is a challenging complication after orthotopic liver transplantation. Its treatment consists of the removal of the precipitating factors. When the etiology is unknown, supportive treatment can be attempted. In severe cases, transjugular intrahepatic portosystemic shunts, portocaval shunts, and liver retransplantation have been used with marginal results. Recently, splenic artery embolization (SAE) has been described as an effective procedure for reducing portal hyperperfusion in patients undergoing partial or whole liver transplantation. Here we describe our experience with SAE for the treatment of RA. Between June 2004 and June 2010, 6 patients underwent proximal SAE for RA. Intraoperative flow measurements, graft characteristics, embolization portal vein (PV) velocities before and after SAE, and spleen/liver volume ratios were collected and analyzed. The response to treatment was assessed with imaging (ultrasound/computed tomography) and on the basis of clinical outcomes (weight changes, diuretic requirements, and the time to ascites resolution). The PV velocity decreased significantly for each patient after the embolization (median = 66.5 cm/second before SAE and median = 27.5 cm/second after SAE, P < 0.01). All patients experienced a significant postprocedural weight loss (mean = 88.1 +/- 28.4 kg before SAE and mean = 75.8 +/- 28.4 kg after SAE, P < 0.01) and a dramatic decrease in their diuretic requirements. All but 1 of the patients experienced a complete resolution of ascites after a median time of 49.5 days (range = 12-295 days). No patient presented with postembolization complications. In conclusion, SAE was effective in reducing the PV velocity immediately after the procedure. Clinically, this translated into a dramatic weight loss, a reduction of diuretic use, and a resolution of ascites. SAE appears to be a safe and effective treatment for RA. PMID- 21618688 TI - The model for end-stage liver disease allocation system for liver transplantation saves lives, but increases morbidity and cost: a prospective outcome analysis. AB - We analyzed the first 100 patients who underwent liver transplantation by Model for End-Stage Liver Disease (MELD) allocation, and compared the outcome of patients on the waiting list and after orthotopic liver transplantation with the last 100 patients who underwent transplantation prior to the introduction of the MELD system in July 2007. MELD allocation resulted in decreased waiting list mortality (386 versus 242 deaths per 1000 patient-years, P < 0.0001) and the transplantation of sicker recipients (uncorrected median MELD score 13.5 versus 20, P = 0.003). Recipient posttransplant morbidity was significantly higher, mainly caused by increased percentage of renal failure requiring renal replacement therapy (13 versus 46%, P < 0.0001). However, kidney function recovered in most cases within 6 months after OLT. Hospital mortality remained similar in both groups (6% versus 9%). Patient 1-year survival was 91% versus 83% (pre-MELD versus MELD era, P = 0.2154), graft 1-year survival was 88% versus 78% (P = 0.1013), respectively. Costs accumulated were significantly higher after introduction of the MELD policy (US $81,967 versus US $127,453, a 55% increase, P = 0.02) with a strong correlation with the individual MELD score (P < 0.0001). The MELD system addresses the goal of fairness well. However, the postoperative course appears more difficult in the MELD era with increased financial burden, but reasonable patient and graft survival. This is the inevitable price to balance justice and utility in liver graft allocation. PMID- 21618689 TI - Explanted liver inflammatory grade predicts fibrosis progression in hepatitis C recurrence. AB - Factors present prior to liver transplantation (LT) that predict fibrosis progression in recurrent hepatitis C infection (HCV) after LT would be important to identify. This study sought to determine if histologic grade of HCV in the explant predicts fibrosis progression in recurrent HCV. The clinical and histologic data of all 159 patients undergoing their first LT for HCV at our center from 1998 to 2001 were retrospectively reviewed with follow-up through June 2008. Twenty-five cases were excluded for: non-HCV-related graft loss <90 days (19), recidivism (4), or unavailable explant or follow-up biopsies (2). A single pathologist scored (Ishak) explants in a blinded fashion. Patients were grouped by explant inflammatory grade <= 4 (group1) and >4 (group 2). Prospectively scored liver biopsies (protocol months 1 and 4, annually, and as indicated clinically) were reviewed for development of advanced fibrosis (bridging or cirrhosis). Cox proportional hazard regression was used to analyze the association of explant grade, donor, viral and LT factors with progression to advanced fibrosis. The groups were well-matched for patient, viral, donor, and transplant factors. Five-year advanced fibrosis-free survival in group 1 versus group 2 was 63% versus 28%, P < 0.001. Explant grade >4 was associated with increased HCV-related graft loss at 1 (6% versus 3%) and 5 (36% versus 14%) years post-LT (P = 0.003). On univariate and multivariate Cox regression analysis, predictors of advanced fibrosis were explant grade >4 (hazard ratio [HR] = 3.3, 95% confidence interval [CI] = 1.9-5.6, P < 0.001) donor age >50 (HR = 3.3, 95% CI = 1.9-5.7, P < 0.001) and viral load at LT of >158,730 IU/mL (HR = 1.8, 95% CI = 1.05-3.1, P = 0.03). CONCLUSION: Explant histologic grade can identify patients requiring more aggressive monitoring and intervention for HCV recurrence post-LT. PMID- 21618690 TI - Preoperative fasting protects mice against hepatic ischemia/reperfusion injury: mechanisms and effects on liver regeneration. AB - We show that brief periods of fasting induce functional changes similar to those induced by long-term dietary restriction in mice, and these changes include protection from ischemia/reperfusion (I/R) injury. In this study, we investigated the mechanisms of protection induced by fasting, and we determined the effect on liver regeneration after partial hepatectomy. Partial hepatic ischemia (75 minutes) was induced in ad libitum fed mice and in 1- to 3-day-fasted mice, and one-third or two-thirds hepatectomy was performed in ad libitum fed mice and 3 day-fasted mice. Preoperative fasting for 2 or 3 days significantly decreased hepatocellular I/R injury. Hepatic gene expression of heme oxygenase 1 (HO-1), superoxide dismutase 2 (SOD2), glutathione peroxidase 1 (Gpx1), and glutathione reductase (GSR) was significantly up-regulated in 3-day-fasted mice at the baseline and 6 hours after reperfusion. After reperfusion, p-selectin and interleukin-6 (IL-6) levels were significantly lower, and superoxide radical generation, lipid peroxidation, and neutrophil influx were significantly attenuated in 3-day-fasted mice. Preoperative fasting did not affect liver regeneration after one-third hepatectomy. Hepatic gene expression of IL-6 and transforming growth factor beta1 was significantly higher in 3-day-fasted mice before and after one-third hepatectomy. Tumor necrosis factor alpha expression significantly increased after one-third hepatectomy in 3-day-fasted mice. After a 3-day fast and two-thirds hepatectomy, liver regeneration and subsequent postoperative recovery were compromised. In conclusion, up-regulation of the stress response gene HO-1 and the antioxidant enzymes SOD2, Gpx1, and GSR at the baseline and a better response after reperfusion likely underlie the protection induced by fasting against hepatic I/R injury. Preoperative fasting may be a promising new strategy for protecting the liver against I/R injury during liver transplantation and minor liver resections, although its effect on extended hepatectomy warrants further exploration. PMID- 21618691 TI - Industrial stabilizers caprylate and N-acetyltryptophanate reduce the efficacy of albumin in liver patients. AB - Liver failure is associated with an accumulation of toxic molecules that exert an affinity to albumin. Some of them have vasoactive activity. So far, albumin has been used as a plasma expander to improve the available circulating blood volume. However, recent studies have suggested that albumin is more effective than starch for this indication. It has not been reported yet whether the industrial stabilizers octanoate and N-acetyltryptophanate, added to albumin, occupy binding sites for vasoactive substances. The aim of this study was to determine whether the presence of the industrial stabilizers octanoate and caprylate has an impact on the effect of the albumin-binding function or circulating blood volume in patients with cirrhosis, portal hypertension, and an indication for albumin. In 25 patients who received albumin via an inline infusion filter that depleted stabilizers, there was an improvement of albumin binding, and there was less deterioration of circulating blood volume and renal function in comparison with a control group. Further studies are needed to confirm the results and identify the potential socioeconomic side effects of industrial stabilizers in commercial albumin solutions. PMID- 21618692 TI - A single-center experience with biliary reconstruction in retransplantation: duct to-duct or Roux-en-Y choledochojejunostomy. AB - Retransplantation is the only therapy for patients who have a failing liver graft, and it can be technically challenging. Although duct-to-duct (DD) biliary reconstruction is considered standard in deceased donor orthotopic whole organ liver transplantation, Roux-en-Y (RY) choledochojejunostomy is preferred by most for biliary reconstruction in retransplantation. We performed a retrospective review of 128 patients who underwent retransplantation after a first transplant with DD biliary construction. Of these 128 patients, 83 had DD biliary reconstructions, and 45 had RY biliary reconstructions. Log-rank tests were used to compare the complication rates between the DD and RY groups, whereas multivariate Cox proportional hazards models were used to compare patient and graft survival between the groups. The median Model for End-Stage Liver Disease score at retransplantation was significantly higher in the DD group (27 versus 21, P = 0.005). The median length of follow-up was 3.3 years. The biliary complication rates were 7% and 11% in the DD group and 10% and 10% in the RY group 30 days and 1 year after retransplantation, respectively (P = 0.73). The rates of primary graft nonfunction complications, hepatic artery thrombosis complications, and reoperation did not differ significantly between groups (all P >= 0.37). In comparison with RY reconstruction, there was no evidence of a difference in patient survival (relative risk = 0.79, P = 0.47) or graft survival (relative risk = 0.94, P = 0.85) for patients with DD reconstruction in multivariate analysis. In conclusion, our results provide evidence that DD biliary reconstruction is feasible in liver retransplantation without increased rates of biliary complications or compromised patient and graft survival. Further studies with larger sample sizes are needed. PMID- 21618693 TI - Graft rotation and late portal vein complications in pediatric living donor liver transplantation using left-sided grafts: long-term computed tomography observations. AB - Right-side rotation of the graft is an uncommon event after pediatric living donor liver transplantation (LDLT) with a left-sided graft. However, graft rotation might lead to gradual portal vein (PV) stretching and late portal vein complications (PVCs). The goal of this study was to quantify the degree of graft rotation (R) by computed tomography (CT) and to determine the effect of graft rotation on the development of late PVCs. One hundred ten patients underwent LDLT with left-sided grafts between 1996 and 2009; CT images were available and were reviewed for 66 of these patients. To quantify R, the following variables were measured with CT: the longest distance between the midline and the extrahepatic PV at the level of the hepatic hilum (A), the distance between the midline and the center of the superior mesenteric vein at the level of the confluence of the splenic vein and superior mesenteric vein (B), and the inner transverse diameter of the body cavity at the level at which A was measured (C). R was calculated as (A - B)/C. In patients with a patent PV (n = 59) and in patients with late PVCs (n = 7), the median R values were 0.16 (range = 0.03-0.38) and 0.25 (range = 0.13 0.39), respectively; there was a significant difference between the 2 groups (P = 0.003). Multivariate analysis showed that R >= 0.2 was the only independent risk factor for the development of late PVCs (P = 0.021). In conclusion, the gradual stretching of the PV after right-side rotation of left-sided grafts might play an important role in the development of late PVCs. PV patency should be closely monitored when graft rotation is noted during clinical follow-up. PMID- 21618695 TI - Abstracts of the 2011 Joint International Congress of ILTS (International Liver Transplantation Society), ELITA (European Liver and Intestine Transplant Association), and LICAGE (Liver Intensive Care Group of Europe). June 22-25, 2011. Valencia, Spain. PMID- 21618694 TI - Noninvasive diagnosis of acute cellular rejection in liver transplant recipients: a proteomic signature validated by enzyme-linked immunosorbent assay. AB - The diagnosis of acute cellular rejection (ACR) requires liver biopsy with its attendant expense and risk. Our first aim was to prospectively determine in an exploratory analysis whether there is a serum proteome signature associated with histologically confirmed ACR. Our second aim was to use simpler and faster enzyme linked immunosorbent assay (ELISA)-based assays for proteins identified as differentially abundant in the proteomic analysis to identify patients with ACR in a separate validation cohort. We used sequential high-abundance protein depletion and isobaric tag for relative and absolute quantitation liquid chromatography-tandem mass spectrometry to characterize the serum proteome in serum samples of patients with or without ACR. Seven of the 41 proteins identified as differentially abundant [serum amyloid A, complement component 4 (C4), fibrinogen, complement component 1q (C1q), complement component 3, heat shock protein 60 (HSP60), and HSP70] could be measured with ELISA-based assays in a validation cohort consisting of patients with ACR (n = 25) and patients without ACR (n = 21). The mean alanine aminotransferase (ALT) levels in patients with ACR and in patients without ACR were 198 +/- 27 and 153 +/- 34 U/L, respectively. Among the 7 proteins for which ELISA assays were available, C4 and C1q were both independent predictors of ACR. C4 had the greatest predictivity for differentiating patients with or without ACR. A C4 level <= 0.31 g/L had a sensitivity of 97%, a specificity of 62%, a positive predictive value of 74%, and a negative predictive value of 94%. A C4 level <= 0.31 g/L and an ALT level >= 70 IU/mL together had a sensitivity of 96%, a specificity of 81%, a positive predictive value of 86%, and a negative predictive value of 94%. In summary, in this exploratory analysis, serum C4 and ALT levels were highly predictive of ACR in liver transplant recipients. Confirmation in a prospective, larger, and diverse population is needed. PMID- 21618696 TI - Transcriptome at the time of hepatitis C virus recurrence may predict the severity of fibrosis progression after liver transplantation. AB - Allograft gene expression analysis may provide insights into the mechanisms involved in liver damage during hepatitis C virus recurrence (HCVrec) after orthotopic liver transplantation (OLT) and allow the identification of patients who have a higher risk of developing severe disease. Forty-three OLT recipients with hepatitis C virus (HCV) were evaluated. Genomewide gene expression analysis was performed with formalin-fixed, paraffin-embedded (FFPE) liver biopsy samples obtained from 21 OLT recipients with HCV at the time of clinical HCVrec, which was defined as increased alanine aminotransferase levels and detectable HCV RNA levels in serum. Patients were classified into 3 groups according to the severity of the fibrosis in the liver biopsies at 36 months post-OLT : group 1 (G1) for mild fibrosis (F0-F1), group 2 for moderate fibrosis (F2), and group 3 (G3) for severe fibrosis (F3-F4). No significant differences were observed between the groups with respect to donor age, histology during HCVrec, treated episodes of acute cellular rejection, or immunosuppression therapy. The results were validated in the remaining 22 OLT recipients with HCV using quantitative real time polymerase chain reaction. Fifty-seven beadtypes showed significantly different expression (P < 0.001) between the groups during HCVrec. In G3, the gene expression of interleukin-28RA (IL-28RA), IL-28, and angiotensin-converting enzyme was up-regulated. Samples from G1 and G3 were used to determine whether a multigenetic classifier could be derived to predict the group class. The final model included the intercept and 9 bead types. Pairwise scatter plots of these 9 bead types revealed that G1 and G3 were well separated with respect to each gene. Our analysis has demonstrated the utility of a set of molecular markers indicating HCVrec severity early after OLT. PMID- 21618697 TI - Liver transplantation for acute intermittent porphyria is complicated by a high rate of hepatic artery thrombosis. AB - Acute intermittent porphyria (AIP) is an autosomal-dominant condition resulting from a partial deficiency of the ubiquitously expressed enzyme porphobilinogen deaminase. Although its clinical expression is highly variable, a minority of patients suffer recurrent life-threatening neurovisceral attacks despite optimal medical therapy. Because the liver is the major source of excess precursor production, liver transplantation (LT) represents a potentially effective treatment for severely affected patients. Using data from the U.K. Transplant Registry, we analyzed all transplants performed for AIP in the United Kingdom and Ireland. Between 2002 and 2010, 10 patients underwent LT for AIP. In all cases, the indication for transplantation was recurrent, biochemically proven, medically nonresponsive acute attacks of porphyria resulting in significantly impaired quality of life. Five patients had developed significant neurological morbidities such as paraplegia before transplantation. The median follow-up time was 23.4 months, and there were 2 deaths from multiorgan failure at 98 days and 26 months. Eight recipients were alive for 3.2 to 109 months after transplantation. Complete biochemical and symptomatic resolution was observed in all patients after transplantation. However, there was a high rate of hepatic artery thrombosis (HAT; 4/10), with 1 patient requiring regrafting. The effects of previous neuronal damage such as joint contractures were not improved by transplantation. Thus, impaired quality of life in the surviving patients was usually a result of preoperative complications. Refractory AIP is an excellent indication for LT, and long-term outcomes for carefully selected patients are good. There is, however, an increased incidence of HAT in these patients, and we recommend routine antiplatelet therapy after transplantation. PMID- 21618698 TI - Safety of liver transplantation with Chagas disease-seropositive donors for seronegative recipients. AB - The shortage of organs for transplantation has prompted the investigation of extended criteria donors, such as donors with transmissible infectious diseases. Here we report our recent experience with liver transplantation using organs from donors who were serologically positive for Chagas disease. We also provide a review of the literature and emphasize donor screening and preventive measures. PMID- 21618704 TI - Renal and urinary proteomics. PMID- 21618705 TI - Distribution of olfactory and nonolfactory surface area in the nasal fossa of Microcebus murinus: implications for microcomputed tomography and airflow studies. AB - The nasal fossa of most mammals exemplifies extreme skeletal complexity. Thin scrolls of bone (turbinals) that both elaborate surface area (SA) and subdivide nasal space are used as morphological proxies for olfactory and respiratory physiology. The present study offers additional details on the nasal fossa of the adult mouse lemur (Microcebus murinus), previously described by Smith and Rossie (Smith and Rossie [2008]; Anatomical Record 291:895-915). Additional, intervening histological sections of the specimen were used to map and quantify the distribution of olfactory and nonolfactory mucosa on the smaller turbinal of the frontal recess (FR; frontoturbinal) and those that occur between ethmoturbinals (ETs; interturbinals). A second adult Microcebus specimen, available as a dried skull, was scanned using microcomputed tomography (microCT) and reconstructed to infer the position of these turbinals within the nasal airway. Overall, turbinal bones comprise more than half of internal nasal SA. All ETs combined comprise about 30% of total nasal fossa SA, and contribute nearly half of all olfactory SA. Of these, the nasoturbinal (NT) is most completely covered with olfactory mucosa, whereas ET I is least covered with olfactory mucosa. The FR contributes significantly to total olfactory SA (ca. 20%). This recess and the single frontoturbinal within it lie in a more lateral pathway of airflow compared with interturbinals, which lie in more central zone just anterior to the olfactory recess of Microcebus. Variations in the turbinals and recesses that complicate central and paranasal in primates should be investigated further in light of zone specific distributions of olfactory receptors (ORs) that differ between these regions in rodents. PMID- 21618706 TI - Multiprotein deposits in neurodegenerative disorders: our experience in the tissue brain bank of Navarra. AB - The prevalence of neurodegenerative disorders increases dramatically with advancing age. Although in recent decades the study of many neurodegenerative disorders has evolved greatly, the concept of neurodegeneration still remains elusive. Although neurodegenerative disorders are classified according to the major components of protein deposits, coexpression of several abnormal proteins in the brain tissue is more common than that was previously thought. The aim of this report is to describe the type of protein deposits found in brains with neuropathological diagnosis of neurodegenerative disease. The report shows the experience obtained in the Brain Bank of Navarra (Spain). The target population for this retrospective descriptive study comprised 178 brains autopsied in the "Hospital of Navarra" in Pamplona between 1994 and 2004 and 201 brains donated to the Brain Bank of Pamplona between 2004 and 2009. The diagnosis of the 201 brains from the Brain Bank was 62 (30.8%) Alzheimer's disease (AD), 43 (21.3%) multiprotein deposit, 31 (15.4%) alpha-synucleinopathies, 31 (15.4%) frontotemporal lobar degeneration (FTLD), 17 (8.4%) tauopathies, 9 (4.4%) prion disease, 6 (2.9%) vascular dementia (VD), and 2 (0.9%) Huntington's disease. Among the 43 cases with multiprotein deposits, we found 35 brains with deposits of 3 proteins (tau, beta-amyloid, and alpha-synuclein). In these two series of brains, the high incidence of deposition of multiple proteins in neurodegenerative disorders is shown. Our results are in agreement with previous findings showing that tau, beta-amyloid, and alpha-synuclein are the proteins most frequently deposited together. PMID- 21618707 TI - The effects of castration and hormone replacement on the cross-sectional area of pubococcygeus muscle fibers in the female rat. AB - In this study, we analyzed the effect of ovariectomy and gonadal hormone replacement on the cross-sectional area of pubococcygeus (Pcm) fibers. It was found that in comparison to intact animals, ovariectomized animals [for 2 or 6 weeks] had an increased cross-sectional area average in Pcm fibers. Ovariectomy also reduced the percentage of fibers with smaller cross-sectional area. In ovariectomized animals after 4 weeks of hormone replacement with an empty Silastic capsule or filled with testosterone propionate or dihydrotestosterone, significantly increased the cross-sectional area average and the percentage of fibers with larger size. However, 17beta-estradiol but not estradiol benzoate treatment reduced the cross-sectional area average and increased the percentage of Pcm fibers with smaller size. Progesterone did not have an effect on the cross sectional area of this muscle. We conclude that Pcm fibers of female rats are sensitive to gonadal hormones, and contrary to male castration, ovariectomy promotes an increase in their cross-sectional area. Also, we discuss according to other studies that an external mechanism which lies within the neuromuscular periphery could also participate in the modulatory hormonal effect on mass or muscle fiber size. Furthermore, in this process, estradiol is likely to regulate the fiber cross-sectional area growing produced by androgens. PMID- 21618708 TI - Upregulation of LAPTM4B-35 promotes malignant transformation and tumorigenesis in L02 human liver cell line. AB - Hepatocellular carcinoma (HCC) is one of the most frequent malignant neoplasms worldwide and is the second leading cause of cancer death in China. We have previously demonstrated that LAPTM4B-35, encoded by lysosomal protein transmembrane 4 beta gene, is overexpressed in over 80% of HCCs and is a novel independent prognostic factor for metastasis, recurrence, and postoperative survival in HCC. In this study, we investigated the role of LAPTM4B-35 in malignant transformation and tumorigenesis using L02 cells, a cell line originated from human normal liver cells. Our data show that replication deficient adenovirus vector-mediated upregulation of LAPTM4B-35 promotes anchorage-independent proliferation and resistance to adriamycin-induced apoptosis. Study of the underlying mechanisms demonstrated alterations of molecular events involved in these processes, which included the activation of phosphoinositide 3-kinases (PI3K)/serine/threonine protein kinase B (PKB/AKT)/bcl xL/bcl-2-associated death promoter homolog (Bad) signaling pathway, inhibition of caspase-3 activation, upregulation of Bcl-2, and downregulation of Bax. In addition, upregulation of LAPTM4B-35 in L02 cells resulted in tumorigenesis in 100% (6/6) of inoculated nude mice and accelerated the death of mice with xenografts in vivo. In conclusion, LAPTM4B-35 promotes malignant transformation and tumorigenesis in human liver L02 cell line through promotion of deregulated proliferation and inhibition of apoptosis. These findings suggest that overexpression of LAPTM4B-35 may play a critical role in hepatocarcinogenesis and therefore, may be a therapeutic target for HCC. PMID- 21618709 TI - Melamed-Wolinska bodies. PMID- 21618710 TI - Adult granulosa cell tumor presenting as metastases to the pleural and peritoneal cavity. AB - Ovarian tumors of sex-cord stromal derivation rarely exfoliate malignant cells in serous effusions. Adult granulosa cell tumor (AGCT) often presents as a stage I disease and association with ascites is about 10%. AGCT is rarely known to exfoliate tumor cells in the ascitic fluid. Exfoliation of malignant cells in the pleural fluid is rarer and has not been described previously in literature to the best of our knowledge. We report a case of AGCT presenting as stage IV disease with exfoliation of malignant cells in the pleural and ascitic fluid with exudative effusions. We wish to emphasize on cytomorphologic features of AGCT to differentiate the same from malignant epithelial tumors. A precise diagnosis in such a case may be possible on serous body fluid cytology with a combined clinical and cytopathologic approach. PMID- 21618711 TI - Periductal mastitis in a male breast masquerading as lobular carcinoma on fine needle aspiration cytology. AB - Fine needle aspiration cytology (FNAC) is quite successful in identifying specific benign and malignant breast lesions, but its role in the categorization of proliferative breast lesions has not been well defined. Mastitis/ breast abscess can lead to intense inflammatory response; however, its association with epithelial hyperplasia is rarely reported. At times, it may be difficult to differentiate dense lymphoplasmacytic infiltrate from non-Hodgkin's lymphoma. We present a case of periductal mastitis associated with epithelial hyperplasia, presenting as a subaerolar swelling in a male breast, which was misinterpreted as lobular carcinoma on FNAC. PMID- 21618712 TI - Inhaled nitric oxide only leads to a transient improvement of oxygenation in patients with acute hypoxemic respiratory failure. PMID- 21618713 TI - Successful prolonged use of recombinant human insulin-like growth factor-1 in a child with cystic fibrosis. AB - Growth failure is a common and complicated process in children with cystic fibrosis (CF). Growth hormone, which is becoming a more commonly used agent in such patients, has demonstrated beneficial effects aside from increased growth velocity. Recently, insulin-like growth factor-1 has gained significant attention in the understanding of growth failure in children with CF. We report the successful prolonged use of recombinant human insulin-like growth factor-1 in an adolescent boy with CF, who demonstrated significant clinical benefits from the therapy. PMID- 21618714 TI - Frequency, treatment, and functional outcome in children with hypersensitivity pneumonitis. AB - BACKGROUND: Hypersensitivity pneumonitis is a rare interstitial lung disease and very few data regarding frequency, treatment and outcome exist for children. Children identified with hypersensitivity pneumonia from a Danish national cohort with diffuse interstitial lung disease form the basis of this study focused on disease frequency, treatment, and functional outcome. METHODS: Seventy-three children with clinical and radiological signs of interstitial lung disease verified by lung biopsy were identified over a 12-year period. Histologic material from all cases was reviewed by pathologists from the ChILD Clinical and Research Network, USA. Diagnosis of hypersensitivity pneumonitis was confirmed in 19 cases. MEASUREMENTS AND MAIN RESULTS: Incidence of hypersensitivity pneumonitis was approximately 2/year and with a point prevalence of 4/1,000,000 children. The median (range) number of monthly courses with intravenous methylprednisolone was 15 courses (8-34) in resolved cases, but in the vast majority (92%), mono-therapy with high dose pulse methylprednisolone treatment was not sufficient for acceptable improvement. Lung function, DLco and DLco/VA increased significantly after 3 and 6 months of treatment compared to baseline (P < 0.05). However, without reaching normal values [mean SDS (range) FEV(1) -0.66 ( 1.88 to 0.41) and FVC -0.67(-1.94 to 0)]. No mortality was seen. CONCLUSIONS: Incidence and point prevalence of hypersensitivity pneumonitis in Denmark was 2/year and 4/1.000.000 children. High dose intravenous methylprednisolone constituted the basic treatment, but in most cases supplemental anti-inflammatory therapy was necessary. Outcome was acceptable without any mortality. Nevertheless, both lung function and diffusion capacity were in subnormal level though without any clinically functional impact. PMID- 21618715 TI - Non-invasive ventilation on a pediatric intensive care unit: feasibility, efficacy, and predictors of success. AB - BACKGROUND: There is only sparse data on the use of non-invasive ventilation (NIV) in acute respiratory failure (ARF) in infants and children. For this setting we investigated feasibility and efficacy of NIV and aimed to identify early predictors for treatment failure. PATIENTS AND METHODS: Retrospective chart review was performed for all patients treated with NIV for ARF from 2003 to 2010 on an 8-bed pediatric intensive care unit of a tertiary university hospital. RESULTS: Seventy-four patients were treated with NIV. One patient did not tolerate mask ventilation and needed immediate invasive ventilation. Intubation rate of the remaining patients was 23% and mortality 15%. Institution of NIV led to significant improvement of both respiratory and heart rate in all patients within the first hour and to further stabilization within the next 8-10 hr. In patients with NIV success blood gases improved significantly 1-2 hr after starting NIV. Multivariate analysis identified low pH after 1-2 hr to be an individual risk factor for NIV failure. Other factors tested were age, underlying disease, acute respiratory insufficiency versus post-extubation failure (PEF), and 1-2 hr after starting NIV oxygen saturation, respiratory rate, PCO(2) , and FiO(2) . Patients with PEF tended to show better outcomes compared to those with acute respiratory insufficiency. CONCLUSION: NIV can be effective in infants and children with ARF. Low pH 1-2 hr after start of NIV is associated with NIV failure. It may therefore be useful in the decision to continue or stop mask ventilation. PMID- 21618716 TI - Use of continuous positive airway pressure (CPAP) in acute viral bronchiolitis: a systematic review. AB - INTRODUCTION: Continuous positive airway pressure (CPAP), used either alone or associated with heliox (CPAP-He), has become a popular therapeutic option for bronchiolitis. This systematic review assesses the impact of CPAP on endotracheal intubation, carbon dioxide pressure (PCO(2) ) and respiratory distress in patients with bronchiolitis. METHODS: Systematic search including studies that used CPAP or CPAP-He in infants with bronchiolitis admitted to a PICU. Data analysis included descriptive statistics and the GRADE system. RESULTS: Five CPAP (one crossover randomized controlled trial [RCT] and four before-after studies) and three CPAP-He (one quasi-RCT and two before-after) studies were included. CPAP was reported to reduce PCO(2) (-6.9 to -11.7 mmHg, respectively, P < 0.015), respiratory rate (-12 to -16 breaths/min after 2 hr, P < 0.01) and the modified Wood clinical asthma score (mWCAS, -2.2 points after 1 hr, P < 0.01). CPAP-He studies observed decreases in PCO(2) (-9.7 mmHg, P < 0.05), mWCAS (-2.12 points, P < 0.001), and respiratory rate (-8 to -13.7 breaths/min, P < 0.05) after 1 hr of treatment. Endotracheal intubation rates ranged from 0-12.5% (CPAP-He) to 17 27% (CPAP). After applying the GRADE system, the quality of evidence for a beneficial effect of CPAP and CPAP-He was classified as low. CONCLUSIONS: The evidence supporting the use of CPAP to reduce PCO(2) and respiratory distress in bronchiolitis is of low methodological quality, and there is no conclusive evidence that CPAP reduces the need for intubation. No definitive conclusions could be drawn about the CPAP-He effect. Further research using higher quality methodology is needed to clarify the beneficial role of these interventions. PMID- 21618717 TI - Surfactant administration prior to one lung ventilation: physiological and inflammatory correlates in a piglet model. AB - OBJECTIVES: To test the hypothesis that surfactant, when given prophylactically during one lung ventilation (OLV), improves physiological stability and reduces inflammation. METHODS: Prospective controlled animal study. After 30 min of mechanical ventilation, surfactant was administered to the left lung of the treatment group. Right lung mechanical ventilation continued for 3 hr, after which the left lung was unblocked. Bilateral mechanical ventilation was continued for 30 min thereafter. Physiological parameters and biomarkers of inflammation in plasma, lung tissue homogenates, and bronchoalveolar lavage (BAL) were measured. MEASUREMENTS AND MAIN RESULTS: Oxygenation improved in the surfactant group, reaching statistical significance at 3 hr of OLV and again after 30 min of bilateral mechanical ventilation following the OLV. Plasma levels of interleukin (IL)-1 beta, IL-6, and tumor necrosis factor (TNF)-alpha showed a trend for reduction. The lung homogenates from the ventilated lungs had significantly lower levels of IL-1 beta (P < 0.01) and IL-6 (P < 0.01). The BAL specimen showed an overall reduction in the cytokine levels; IL-1 beta was significantly lower in the ventilated lungs (P < 0.01). CONCLUSIONS: Surfactant administration improves oxygenation and decreases inflammation, as evidenced by a decrease in several inflammatory cytokines both in the plasma and lungs of a piglet model of OLV. PMID- 21618718 TI - Pulmonary maturational arrest and death in a patient with pulmonary interstitial glycogenosis. AB - We present the clinical presentation and pathological findings from a term infant with atypical neonatal lung disease. A full term Caucasian male presented at birth with signs of respiratory distress. The respiratory condition continued to deteriorate despite maximum intervention and the patient was placed on ECMO for further cardiorespiratory assistance. An open lung biopsy demonstrated findings consistent with severe lung growth abnormality with non-uniform pulmonary interstitial glycogenosis. The patient consequently developed a pulmonary hemorrhage that required discontinuation of ECMO. The patient died shortly after decannulation. Most literature suggests that PIG is one of the few pediatric interstitial lung diseases that has a favorable prognosis with rare mortality in the absence of co-morbidities. However, the current case suggests prognosis may depend more on the underlying diagnosis than on the histological finding of PIG. In addition, this case may provide insight into the pathogenesis and potential modifiers of this idiopathic disorder. PMID- 21618719 TI - Chest radiographic and CT evaluation of lung abnormalities in pediatric patients with laryngeal cleft. AB - BACKGROUND: The purpose of this study was to (1) evaluate chest radiography (CR) and computed tomography (CT) findings in pediatric patients with laryngoscopically confirmed laryngeal cleft and (2) determine whether CT provided additional information over CR in evaluating lung abnormalities in pediatric patients with laryngeal cleft. METHODS: Two pediatric radiologists performed a retrospective review of CRs and CTs in pediatric patients with laryngoscopically confirmed laryngeal cleft from January, 2002 to January, 2010. Lungs were evaluated for pattern (airspace or interstitial), distribution (upper, middle, or lower lung zones), and extent (<25% or >25%) of abnormalities on CRs and CTs. In patients who had both CR and CT, CT findings were compared with concurrent CRs to determine whether CT provided additional information and resulted in alterations in patient management. RESULTS: The final study cohort consisted of 67 pediatric patients (37 males and 30 females; mean age 3.0 years +/- 3.5 years; range 25 days to 17.5 years) who had CR. Sixteen (24%) of these 67 patients had both CR and CT. Chest radiographs were abnormal in 47 patients (70%) which included consolidation (n = 35; 74%) and reticular opacities in 12 patients (26%). Lung parenchymal abnormalities were observed in the upper (n = 10; 21%), middle (n = 2; 4%), or lower (n = 35; 75%) lung zones on CR. The extent of lung parenchymal abnormalities was >25% in 30 patients (64%) and <25% in 17 patients (36%). Chest CT examinations were abnormal in all 16 patients (100%) who had available chest CT examinations, which included consolidation (n = 9; 56%) and reticular opacities (n = 7; 44%). Lung parenchymal abnormalities were observed in the upper (n = 2; 12.5%) or lower (n = 14; 87.5%) lung zones on CT. The extent of lung abnormalities was >25% in 10 patients (60%) and <25% in six patients (40%). CT provided major additional findings in 12 patients (75%) and minor findings in four patients (25%). Additionally, CT improved the evaluation of the distribution and extent of lung abnormalities in 11 patients (69%). The additional information provided by CT resulted in alteration of patient management in seven patients (43.8%). CONCLUSIONS: Among pediatric patients with laryngeal cleft, CR and CT often show lung abnormalities such as consolidation and reticular opacities. CT frequently provides additional information, which may change patient management. PMID- 21618720 TI - Is aspiration during swallowing more common in Canadian children with indigenous heritage? AB - While many factors that contribute to the occurrence of lower respiratory tract infections (LRTI) in Canadian indigenous children have been described, the role of aspiration during swallowing has not been explored in these children. Because of an increase in referrals of indigenous children from our catchment area (Manitoba, North Western Ontario, and Nunavut) for assessment of aspiration during swallowing, we retrospectively reviewed the clinical records of 325 consecutive children undergoing videofluoroscopic swallowing studies (VFSS) to evaluate which factors, including indigenous heritage and LRTI, were associated with aspiration during swallowing. Our sample had an overrepresentation of indigenous children (35% compared to 23% in the catchment area). These children were more likely to aspirate during swallowing (P = 0.001) and to have experienced an LRTI prior to the VFSS (P = 0.000). When separating the children who aspirated into two groups based on indigenous heritage, the indigenous children were more likely to have had an LRTI (P = 0.028) than the other children in the sample. With logistic regression analysis, significant correlations between indigenous heritage, LRTI (P = 0.000), and aspiration (P = 0.009) were found. When aspiration during swallowing was the factor of interest, it correlated with both LRTI (P = 0.001) and the presence of congested upper airway noises after eating (P = 0.000). These upper airway noises were strongly associated with aspiration in indigenous children (P = 0.004). While prospective data are required to determine if the correlations seen in this retrospective review will remain robust, indigenous children were more likely to aspirate during swallowing and have LRTI in this sample. Whether aspiration during swallowing is related to environmental, ethnic, or biological factors in indigenous children remains to be determined, but the association between aspiration and congested upper airway noises after eating may assist in the early diagnosis of aspiration during swallowing, and thereby enable timely intervention to decrease aspiration risk in children already at risk of repeated LRTI due to a number of other confounding factors. PMID- 21618722 TI - In vitro cell culture pO2 is significantly different from incubator pO2. AB - Continuous noninvasive monitoring of peri-cellular liquid phase pO2 in adherent cultures is described. For neurons and astrocytes, this approach demonstrates that there is a significant difference between predicted and observed liquid phase pO2. Particularly at low gas phase pO2s, cell metabolism shifts liquid phase pO2 significantly lower than would be predicted from the O2 gas/air equilibrium coefficient, indicating that the cellular oxygen uptake rate exceeds the oxygen diffusion rate. The results demonstrate the need for direct pO2 measurements at the peri-cellular level, and question the widely adopted current practice of relying on setting the incubator gas phase level as means of controlling pericellular oxygen tension, particularly in static culture systems that are oxygen mass transfer limited. PMID- 21618721 TI - Hypoxia and nitric oxide exposure promote apoptotic signaling in contractile pulmonary arterial smooth muscle but not in pulmonary epithelium. AB - RATIONALE: Neonatal pulmonary hypertension is characterized by hypoxia, abnormal vascular remodeling, and impaired alveolarization. Nitric oxide (NO) regulates cell replication and activation of apoptosis. Our objective was to examine cell phenotype-specific effects of hypoxia and NO exposure on cumulative apoptotic signal in neonatal pulmonary epithelial cells and arterial smooth muscle. DESIGN/METHODS: Primary cultured newborn porcine pulmonary arterial myocytes and epithelial cells were grown in normoxic (21% O2) or hypoxic conditions (10% O2). Myocyte phenotype was predetermined by serum-supplementation or -deprivation. Cells were exposed to sodium nitroprusside (10(-7) -10(-4) M) or diluent for 3 days. Cell survival was estimated by MTT assay; BAX, Bcl-2, and cleaved caspase-3 by Western blot; cell cycle entry by laser scanning cytometry. RESULTS: Hypoxic epithelial cells exhibited a small increase in anti-apoptotic Bcl2, and decrease in BAX. Cell survival and active caspase-3 were unchanged. Exposure to NO had no impact on epithelial apoptosis, but initiated necrosis. In contractile myocytes, pro-apoptotic BAX abundance and caspase-3 activation were increased by hypoxia, augmented by NO exposure promoting apoptosis. Hypoxia decreased BAX/Bcl-2 ratio and promoted survival of synthetic myocytes; NO increased apoptosis of normoxic synthetic myocytes, but decreased apoptosis of hypoxic synthetic myocytes. CONCLUSION: The effect of NO on pulmonary apoptosis is phenotype-dependent. A cumulative apoptotic effect of hypoxia and NO in vitro exerted on contractile myocytes may lead to contraction of this subpopulation, while synthetic myocyte survival and proliferation is enhanced by hypoxia and NO. Epithelial survival is unaffected. We speculate that alveolar rarefaction reported after neonatal hypoxia may arise from growth arrest in the vascular rather than the epithelial compartment. PMID- 21618723 TI - CFD-aided cell settler design optimization and scale-up: effect of geometric design and operational variables on separation performance. AB - The inclined multiplate (lamella) gravity settler has proven to be an effective cell retention device in industrial perfusion cell culture applications. Investigations on the effects of geometric design and operational variables of the cell settler are crucial to understanding how to best improve the settler performance. Maximizing the harvest/perfusion flow rate while minimizing viable cell loss out of the harvest is the primary challenge for optimization of the settler design. This study demonstrated that computational fluid dynamics (CFD) can be utilized to accurately model and evaluate the settler separation performance for near-monodisperse suspensions and therefore aid in the design optimization of the settler under these baseline conditions. With the preferred geometric features that were identified from CFD modeling results, we proposed design guidelines for the scale-up of these multiplate settler systems. With these guidelines and performance verification using the CFD model, a new large scale settler was designed and fabricated for a perfusion cell culture process using a minimally aggregating production cell line. Perfusion cell culture runs with this particular cell line were performed with this settler, and the CFD model was able to predict the initial ramp-up performance, proving it to be a valuable scale-up design tool for this production process. PMID- 21618724 TI - Predicting Mab product yields from cultivation media components, using near infrared and 2D-fluorescence spectroscopies. AB - The yield of monoclonal antibody (Mab) production processes depends on media formulation, inocula quality, and process conditions. As in industrial processes tight cultivation conditions are used, and inocula quality and viable cell densities are controlled to reasonable levels, media formulation and raw materials lot-to-lot variability in quality will have, in those circumstances, the highest impact on process performance. In the particular Mab process studied, two different raw materials were used: a complex carbon and nitrogen source made of specific peptones and defined chemical media containing multiple components. Using different spectroscopy techniques for each of the raw material types, it was concluded that for the complex peptone-based ingredient, near-infrared (NIR) spectroscopy was more capable of capturing lot-to-lot variability. For the chemically defined media containing fluorophores, two-dimensional (2D) fluorescence spectroscopy was more capable of capturing lot-to-lot variability. Because in Mab cultivation processes both types of raw materials are used, combining the NIR and 2D-fluorescence spectra for each of the media components enabled predictive models for yield to be developed that out-performed any other model involving either one raw material alone, or only one type of spectroscopic tool for both raw materials. For each particular raw material, the capability of each spectroscopy to detect lot-to-lot differences was demonstrated after spectra preprocessing and specific wavelength regions selection. The work described and the findings reported here open up several possibilities that could be used to feed-forward control the process. These include, for example, enabling specific actions to be taken regarding media formulation with particular lots, and all types of predictive control actions aimed at increasing batch-to-batch yield and product quality consistency at harvest. PMID- 21618725 TI - Process and economic evaluation for monoclonal antibody purification using a membrane-only process. AB - In recent years, the market for therapeutic monoclonal antibodies (mAb) has grown exponentially, and with this there has been a desire to reduce the costs associated with production and purification of these high-value biological products. A typical mAb purification process involves three adsorption/chromatography steps [protein A, ion exchange (IEX), and hydrophobic interaction (HIC)], along with ultrafiltration, nanofiltration, and microfiltration. With the development of membrane adsorption/chromatography as a viable alternative to traditional pack bed systems, the opportunity exists to complete the entire downstream purification process using only membrane operations. In this study, the process simulation tool SuperPro Designer was used to evaluate the application of recently developed ultra-high capacity electrospun nanofibrous adsorption membranes as a replacement for conventional chromatographic media in the downstream mAb production process. The simulation showed that nanofibrous adsorption membranes in place of the three packed bed chromatography steps reduced the required volume of protein A, IEX, and HIC adsorptive medium by 25, 80, and 80%, respectively. In addition, the membrane only process reduced the downstream processing time by 50%, decreased the number of labor hours associated with the purification steps by 40%, generated 40% less aqueous waste, and reduced the overall downstream process operating expenses per unit product by 23%. There were also significant savings in facility construction costs and the price of fixed equipment required for separations. With these savings not only is the membrane-only process economically competitive with the traditional packed bed operations, but it offers the possibility of moving toward more disposable process. PMID- 21618726 TI - Commentary on Liang & Chikritzhs (2011): Quantifying the impacts of health problems on drinking and subsequent morbidity and mortality - life-course measures are essential. PMID- 21618727 TI - Ophthalmological numismatics. Antonio Scarpa (1752-1832). PMID- 21618729 TI - Abstracts of the European Academy of Childhood Disability 23rd Annual Meeting. June 8-11, 2011. Rome, Italy. PMID- 21618728 TI - [Information included in the summary of product characteristics of the radiopharmaceuticals for human use approved for marketing in Spain]. PMID- 21618730 TI - Commentary on Lund et al. (2011): Consolidating the evidence on effectiveness of snus for smoking cessation - implications for public health. PMID- 21618731 TI - Regenerative medicine spoilt for choice. PMID- 21618732 TI - Tony Hyman. PMID- 21618733 TI - Adult somatic stem cells: new perspectives. Proceedings of the 2nd International Congress on Responsible Stem Cell Research. Monaco. November 26-28, 2009. PMID- 21618734 TI - Proceedings of the First World Congress of Pediatric Urology. May 27-31, 2011. San Francisco, California, USA. PMID- 21618735 TI - Proceedings of the 11th Annual Bioinformatics Open Source Conference (BOSC) 2010. July 9-10, 2010. Boston, Massachusetts, USA. PMID- 21618736 TI - Proceedings of the 62nd International Symposium on Crop Protection. May 18, 2010. Gent, Belgium. PMID- 21618737 TI - Proceedings of 2010 Perfusion Downunder Winter Meeting. August 5-8, 2010. Queenstown, New Zealand. PMID- 21618738 TI - Critical condition: integrated health care to increase quality of life and life expectancy. Proceedings of an integrated health care conference. Chicago, Illinois, USA. 2009. PMID- 21618739 TI - Reflections: Neurology and the humanities. Reflections for April: The silent witness and Charcot's hat. PMID- 21618740 TI - Immune reconstitution inflammatory syndrome after withdrawal of natalizumab? PMID- 21618741 TI - Microporous organic polymers: design, synthesis, and function. AB - Microporous organic polymers (MOPs) can be defined as materials with pore sizes smaller on average than 2 nm which are comprised of light, non-metallic elements such as C, H, O, N, and B. We describe here the main classes of MOPs which are conveniently sub-divided into amorphous and crystalline groups. We present an overview of the synthesis of these materials, along with some general design criteria for producing MOPs with high surface areas and micropore volumes. The advantages and disadvantages of MOPs with respect to inorganic materials such as zeolites and hybrid materials such as metal organic frameworks are discussed throughout, particularly in terms of practical applications such as catalysis, separations, and gas storage. We also discuss future opportunities in this area as well as the potential to unearth "undiscovered" MOPs among the large number of rigid backbone polymers and networks reported in the literature. PMID- 21618742 TI - Hydrogen, methane and carbon dioxide adsorption in metal-organic framework materials. AB - The role of porous coordination polymers (metal-organic frameworks) as storage materials for hydrogen, carbon dioxide and methane and the current state-of-the art in this rapidly developing field are reviewed. The significant potential of porous materials to store fuel gases effectively and reversibly illustrates the great promise of engineered solid-state materials as the next generations of applied multifunctional materials. PMID- 21618743 TI - Doping of metal-organic frameworks with functional guest molecules and nanoparticles. AB - Nanoparticle synthesis within metal-organic frameworks (MOFs) is performed by the adsorption of suitable precursor molecules for the metal component and subsequent decomposition to the composite materials nanoparticles@ MOF. This chapter will review different approaches of loading MOFs with more complex organic molecules and metal-organic precursor molecules. The related reactions inside MOFs are discussed with a focus on stabilizing reactive intermediates in the corresponding cavities. The syntheses of metal and metal oxide nanoparticles inside MOFs are reviewed, and different synthetic routes compared. Emphasis is placed on the micro structural characterization of the materials nanoparticles@MOF with a particular focus on the location of embedded nanoparticles using TEM methods. Some first examples of applications of the doped MOFs in heterogeneous catalysis and hydrogen storage are described. PMID- 21618744 TI - Chiral metal-organic porous materials: synthetic strategies and applications in chiral separation and catalysis. AB - In the light of growing demand for chiral purity in biological and chemical processes, the synthesis of chiral metal-organic porous materials (CMOPMs) have become immensely important because of their potential applications in chiral separation and asymmetric catalysis. In this chapter, the synthetic strategies for CMOPMs are discussed briefly keeping the focus on their applications. Two distinct approaches have been taken to synthesize a wide variety of chiral structures with different topologies and accessible cavities. Several CMOPMs have shown catalytic activity and enantioselectivity toward a number of chemical transformations. On many occasions, the chiral pores of the MOPMs have been utilized in order to achieve separation of enantiomers from racemates. Recent applications of homochiral MOPMs in heterogeneous asymmetric catalysis and chiral separations are also presented here. PMID- 21618745 TI - Controlled polymerization by incarceration of monomers in nanochannels. AB - Porous Coordination Polymers (PCPs) composed of transition metal ions and bridging organic ligands have been extensively studied. The characteristic features of PCPs are highly regular channel structures, controllable channel sizes approximating molecular dimensions, designable surface potentials and functionality, and flexible frameworks responsive to guest molecules. Owing to these advantages, successful applications of PCPs range from molecular storage and separation to heterogeneous catalysts. In particular, use of their regulated and tunable nanochannels in the field of polymerization has allowed multi-level control of polymerization via control of stereoregularlity, molecular weight, etc. In this chapter, we focus on recent progress in polymerization utilizing the nanochannels of PCPs, and demonstrate why this polymerization system is attractive and promising from the viewpoint of precision control of polymeric structures. PMID- 21618746 TI - Designing metal-organic frameworks for catalytic applications. AB - Metal-organic frameworks (MOFs) are constructed by linking organic bridging ligands with metal-connecting points to form infinite network structures. Fine tuning the porosities of and functionalities within MOFs through judicious choices of bridging ligands and metal centers has allowed their use as efficient heterogeneous catalysts. This chapter reviews recent developments in designing porous MOFs for a variety of catalytic reactions. Following a brief introduction to MOFs and a comparison between porous MOFs and zeolites, we categorize catalytically active achiral MOFs based on the types of catalytic sites and organic transformations. The unsaturated metal-connecting points in MOFs can act as catalytic sites, so can the functional groups that are built into the framework of a porous MOF. Noble metal nanoparticles can also be entrapped inside porous MOFs for catalytic reactions. Furthermore, the channels of porous MOFs have been used as reaction hosts for photochemical and polymerization reactions. We also summarize the latest results of heterogeneous asymmetric catalysis using homochiral MOFs. Three distinct strategies have been utilized to develop homochiral MOFs for catalyzing enantioselective reactions, namely the synthesis of homochiral MOFs from achiral building blocks by seeding or by statistically manipulating the crystal growth, directing achiral ligands to form homochiral MOFs in chiral environments, and incorporating chiral linker ligands with functionalized groups. The applications of homochiral MOFs in several heterogeneous asymmetric catalytic reactions are also discussed. The ability to synthesize value-added chiral molecules using homochiral MOF catalysts differentiates them from traditional zeolite catalysis, and we believe that in the future many more homochiral MOFs will be designed for catalyzing numerous asymmetric organic transformations. PMID- 21618747 TI - Magnetic and porous molecule-based materials. AB - In this chapter, we give an overview of the recent state-of-the-art research of porous and magnetic molecule-based materials. The subject is introduced by a section devoted to the fundamentals of magnetism in molecular magnets, with special attention to the design strategies to prepare molecular magnetic materials. We will then focus on the two main families of materials combining porosity and magnetism: the purely organic and the metal-organic porous magnetic materials. For both families, a selection of the most representative examples has been made. A complete section is devoted to magnetic and porous materials with flexible frameworks, an area of emerging importance in this field, because of their wide range of applications. Finally, we conclude with a brief overview on the most recent approaches for the future development of these materials. PMID- 21618748 TI - Acute leukemia and aggressive lymphoma treatment in adults: it is time for Malawi to move forward. PMID- 21618749 TI - Au contraire: response to "Neither/nor". PMID- 21618750 TI - Commentary on Kuhns et al. (2011): Drinks and death - alcohol and the homicide victim. PMID- 21618751 TI - Lab personnel. Technically gifted. PMID- 21618752 TI - Commentary on Van Rooij et al. (2011):'Gaming addiction' - a psychiatric disorder or not? PMID- 21618753 TI - Commentary on Caiaffa et al. (2011):The renewed challenge of hepatitis C virus epidemiology among non-injecting drug users. PMID- 21618754 TI - Abstracts of the 46th Congress of the European Society for Surgical Research. Aachen, Germany. May 25-28, 2011. PMID- 21618755 TI - [Mediator: a French problem]. PMID- 21618756 TI - [No, palliative care is not a loss of opportunity!]. PMID- 21618757 TI - [Your images. Pneumomediastinum]. PMID- 21618758 TI - [Age related macular degeneration]. AB - Age-related macular degeneration (ARMD) is a multifactorial disease caused by a combination of genetic and environmental factors. It is the first cause of blindness in patients over 50 in the western world. The disease has been traditionally classified into early and late stages with dry (atrophic) and wet (neovascular) forms: neovascular form is characterized by new blood vessels development under the macula (choroidal neovascularisation) which lead to a rapid decline of vision associated with metamorphopsia and requiring an urgent ophtalmological examination. Optical coherence tomography is now one of the most important part of the examination for diagnosis and treatment. Patient with age related maculopathy should consider taking a dietary supplement such that used in AREDS. The treatment of the wet ARMD has largely beneficied since year 2006 of anti-VEGF (vascular endothelial growth factor) molecules such as ranibizumab or bevacizumab given as repeated intravitreal injections. A systematic follow up each 4 to 8 week in required for several years. There is no effective treatment at the moment for dry AMD. For patients with binocular visual acuity under 60/200 rehabilitation includes low vision specialist, vision aids and psychological support. PMID- 21618759 TI - [Gaucher disease]. PMID- 21618760 TI - [Suicide prevention: caring for others]. PMID- 21618761 TI - [Suicide: epidemiological data]. AB - Within developed countries suicide rates in France are one of the highest, and are a matter of public health concern. In 2008 (latest available year) suicide figure is 10,313 (7,589 men and 2,724 women), which represents rates of 25.2 for men and 8.5 for women (16.6 p.100,000 persons). About 20% of suicide death are not registered. Suicide has increased since the mid 70'until the mid 80' and has declined since that period, but remains still higher than before for women (about 5% more). For baby-boomers the increase of suicide with age is more important until 50 years old, but seems less after. This new age-trend is becoming the same as in northern Europe and western countries. Suicide is prevalent by the age of 30 years old: suicide death is the first cause of death by that age, and suicide attempts are at the top incidence rate. Suicide is an important cause of premature mortality, and attempted suicide a frequent cause for hospitalization especially for young people (more than 180,000 each year, all ages). According to way of live, those who are alone are more at risk, especially the widowers. Mental diseases are prevalent in past life for suicide patients. Risk for those persons is higher, and each year about 1% of suicide attempters make suicide. PMID- 21618762 TI - [Werther effect and suicide contagion]. PMID- 21618763 TI - [Risk factors of suicide]. PMID- 21618764 TI - [Suicidal crisis]. AB - The different steps of the suicidal crisis are described according to the recommendations of the French consensus conference. The point of view of the patient is developed in a psychodynamic perspective. Three clinical forms are distinguished according to the causes network. Are described the main protective factors for the elderly because suicide is a major public health concern for older adults in France. PMID- 21618765 TI - [Detecting the alarm signs of the suicidal crisis]. AB - Suicidal crisis is an escaping moment during which the subject shows a state of vulnerability and a lack of his/her defense abilities, putting him/her in a pain which is not always visible and that does not express itself through a plain nosographic context. Because of that, the practitioner's intervention is made more complicated. In this article, we offer a way to conduct the interview to detect the alarm signs of the suicidal crisis, insisting on the clinical, biographical and environmental elements that are to be looked for. Therefore, the analysis of the suicidal discourse (suicidal ideas expressed or hidden suicidal complaints), the search for risk and protective factors clearly identified, and the interest shown to the personal experiment of the crisis state, will make the evaluation and the determination of suicidal potentialities easier. This is based on the tripod Risk-Emergency-Dangerousness (RED), which will give a direction to the most efficient 24-48 hours care at the conclusion of the interview, whether it were ambulatory (low suicidal potentiality) or requiring an hospitalization (medium or high suicidal potentiality). PMID- 21618766 TI - [Suicidal crisis: when to hospitalize and how to treat?]. PMID- 21618767 TI - [Suicidal behavior in children]. AB - Because of the rarity of suicide and attempted suicide in young children the subject is not commonly studied and is quite poorly understood. Indeed, we have longed denied the possibility of a young child's enactment of a suicidal desire. The myth of total childhood innocence paired with societal views on death and mourning make suicide at this age unacceptable and furthermore, unthinkable. Everything is positioned towards denial; however suicidal behaviour does exist in children and we can identify specifics surrounding the methods used, and their meaning in both the social and environmental contexts in which they take place. Within the following we will suggest and provide tools for prevention and treatment. PMID- 21618768 TI - [Suicide and work]. AB - The occurrence of suicide series in the professional setting seems to be a relatively recent phenomenon, even though past and current epidemiological data are lacking to assess their progression. Causes of suicidal gesture are subject to recurrent controversies. Contradictory theories (structuralist theories vs sociogenetic theories) should be overcome to explore relationships between subjects and their work environment. Most of the time, subjects share two features: loneliness and loss of acting power. Diseases of excited activity or impeded activity shed light on the motivations that may lead to rupture. PMID- 21618769 TI - [What can we do to prevent the suicide re-attempts?]. AB - A subject surviving a suicide attempt (SA) belongs in fact to a group at risk for suicide (40% of lifetime repetition including 20 to 25% over the 12 months following the initial gesture). To prevent the risk of suicide in general is thus effective on the prevention of the repetition. It initially seems important to treat a somatic or psychiatric pathology having taken part in the initial suicidal context: treating a mood depression disorder, prescribing a mood stabilizer to a bipolar patient, managing the global treatment of a borderline personality disorder, etc. Some strategies have been proposed with a specific aim to reduce this rate of suicidal repetition. Certain devices very interventionists appear expensive and difficult to generalize (at home interventions, intensive short psychotherapies carried out starting from the Emergency Rooms...). In a parallel way, "connectedness" devices, which are careful not to invade the suicide attempter life, which does not aim to replace a treatment, but try to propose effective recourse in case of crisis, tend to currently develop on the whole territory. PMID- 21618770 TI - [Drug and suicide risk]. PMID- 21618771 TI - [Acute pancreatitis: positive and etiological diagnosis, progressive forms]. AB - The diagnosis of acute pancreatitis is based on the association of an acute abdominal pain with raised level of seric lipase exceeding three fold the normal value during the first 48 h following pain occurrence. The main causes are biliary lithiasis and chronic alcoholism. Oedematous pancreatitis resume in a few days whereas severe necrotizing pancreatitis may require long staining and complex medico-surgical management. PMID- 21618772 TI - [Pathophysiology and cause of acute pancreatitis]. PMID- 21618773 TI - [Imaging of acute pancreatitis]. AB - The radiological management of acute pancreatitis is well codified. The positive diagnosis of acute pancreatitis is based on clinical and biological signs. However, imaging helps for in diagnosing severity and complications. The etiological diagnosis should strive to detect gallstones by an abdominal ultrasound to be performed within 24 hours of onset of symptoms. The severity is assessed at 48-72 hours per scanner after contrast media injection, according to the classification CTSI. It is important to avoid the two following pitfalls: realising the scanner too early, which could be falsely reassuring because necrosis of the pancreatic parenchyma appears at least 48 to 72 hours after onset of clinical signs, and dispense with ultrasound at 24 hours, because the gallstones are visible only in 50% of the scanner. Imaging helps to manage some complications of acute pancreatitis: in case of infectious complications, remember to perform a needle aspiration or percutaneous drainage of fluid collections; in case of haemorrhage from rupture of a false aneurysm, the treatment of choice is embolization for hemostasis under radiological guidance. PMID- 21618774 TI - [Severity assessment and medical treatment of acute pancreatitis]. AB - Mortality rate of acute pancreatitis is 5% but can reach 47% in necrotizing pancreatitis with multiorgan failure. Prediction of an unfavourable outcome is based on bioclinical scoring systems either specific of pancreatitis or not, single biological prognostic criteria as CRP and procalciton levels measured during the first 48 h and abdominal CT-scan. Medical treatment of acute pancreatitis is symptomatic and requires intensive care in severe forms of the disease. PMID- 21618775 TI - [Treatment of gallstone disease responsible for acute pancreatitis]. PMID- 21618776 TI - [Surgical treatment of severe acute pancreatitis]. AB - Necrosectomy and drainage, mostly surgically performed, are indicated in front of superinfection of pancreatic necrosis proven by percutaneous bacteriological puncture or when the clinical status of the patient deteriorates despite maximal intensive care, without proven infection of necrosis. Early necrosectomy (within the first two or three weeks) carries a risk of postoperative mortality around 50%. To delay further the necrosectomy is advisable when it is compatible with patient's status. PMID- 21618777 TI - [Minimally invasive procedure in pancreatic necrosis]. PMID- 21618778 TI - [Rank examination and residencies: some common ideas]. PMID- 21618779 TI - [Tetanus: the necessity of maintaining an optimal coverage]. PMID- 21618780 TI - [Migraine and cluster headache]. PMID- 21618781 TI - [Breastfeeding: what information do we need to give to expectant mothers?]. PMID- 21618782 TI - [Accidents caused by anticoagulants]. PMID- 21618783 TI - [Extra-uterine pregnancy]. PMID- 21618784 TI - [Normal aging: biological, functional and relational aspects. Epidemiological and sociological data. Prevention of pathological aging]. PMID- 21618785 TI - [Mumps]. PMID- 21618786 TI - [Hypoglycemia: different etiologies with and without a history of diabetes]. PMID- 21618787 TI - [Jean Fernel, humanist and physician]. PMID- 21618788 TI - [Overcoming intractable diseases. Preface]. PMID- 21618789 TI - [Parkinson disease/Parkinsonism]. PMID- 21618790 TI - [Current status and prospective of pituitary disease]. PMID- 21618791 TI - [Idiopathic cardiomyopathy]. PMID- 21618792 TI - [Pulmonary hypertension]. PMID- 21618793 TI - [Progress and achievement of rare/intractable diseases research in Japan]. PMID- 21618794 TI - [Current status of diagnosis and treatment for antineutrophil cytoplasmic antibody associated vasculitis]. PMID- 21618795 TI - [Idiopathic bone marrow disorders--focusing on the treatment of MDS]. PMID- 21618796 TI - [Refractory nephrotic syndrome, perspectives of treatment]. PMID- 21618797 TI - [Current state of fulminant hepatitis and countermeasure for de novo B hepatitis]. PMID- 21618798 TI - [The 38th Scientific Meeting: Perspectives of internal medicine. Summary]. PMID- 21618799 TI - [Current state and the future of gastro-intestinal disease]. PMID- 21618800 TI - [Lifestyle-related liver diseases]. PMID- 21618801 TI - [Current status and future prospects of diabetes therapy]. PMID- 21618803 TI - [Progress in acute and chronic stages of ischemic stroke]. PMID- 21618802 TI - [Clinical guideline for thyroid disease]. PMID- 21618804 TI - [Practical diagnosis and treatment for dementia]. PMID- 21618805 TI - [Early detection and treatment of rheumatoid arthritis]. PMID- 21618806 TI - [Systemic lupus erythematosus--recent progress on diagnosis and therapy]. PMID- 21618807 TI - [An approach to vertigo for general internist in emergency room]. PMID- 21618808 TI - [Assessment and treatment of coronary artery disease: present status and future perspectives]. PMID- 21618809 TI - [Use of anticoagulant and antiplatelet in general practice]. PMID- 21618810 TI - [Recent progress for chemotherapy in advanced non-small cell lung cancer]. PMID- 21618811 TI - [Diagnosis and treatment of bronchiolitis obliterans]. PMID- 21618812 TI - [Diagnosis and treatment for patients with chronic kidney disease]. PMID- 21618813 TI - [Hypertension and chronic kidney disease]. PMID- 21618814 TI - [Recent advances in the diagnosis and treatment of multiple myeloma]. PMID- 21618815 TI - [Molecular pathogenesis and treatment of leukemia]. PMID- 21618816 TI - [Recent epidemiology and therapy of HIV infection]. PMID- 21618817 TI - Health care costs in America-technology as a major driver. PMID- 21618818 TI - [Diseases and ion dynamics of intracellular organelles as the target of drug designing. Preface]. PMID- 21618819 TI - [Chronic noncancer pain and opioids]. PMID- 21618820 TI - [Revitalization, reconstructive or more efficiency in research activities at medical schools?]. PMID- 21618821 TI - Adhesion-GPCRs: structure to function. Preface. PMID- 21618822 TI - The adhesion GPCRs; gene repertoire, phylogeny and evolution. AB - The Adhesion family is unique among the GPCR (G protein-coupled receptor) families because of several features including long N-termini with multiple domains. The gene repertoire has recently been mined in great detail in several species including mouse, rat, dog, chicken and the early vertebrate Branchiostoma (Branchiostoma floridae) and one of the most primitive animals, the cniderian Nematostella (Nematostella vectensis). There is a one-to-one relationship of the rodent (mouse and rat) and human orthologues with the exception the EMR2 and EMR3 that do not seem to have orthologues in either rat or mouse. All 33 human Adhesion GPCR genes are present in the dog genome but the dog genome also contains 5 additional full-length Adhesion genes. The dog and human Adhesion orthologues have higher average protein sequence identity than the rodent (rat and mouse) and the human sequences. The Adhesion family is well-represented in chicken with 21 one-to-one orthologous with humans, while 12 human Adhesion GPCRs lack a chicken ortholog. Branchiostoma has rich repertoire of Adhesion GPCRs with at least 37 genes. Moreover, the Adhesion GPCRs in Branchiostoma have several novel domains their N-termini, like Somatomedin B, Kringle, Lectin C-type, SRCR, LDLa, Immunoglobulin I-set, CUB and TNFR. Nematostella has also Adhesion GPCRs that are show domain structure and sequence similarities in the transmembrane regions with different classes of mammalian GPCRs. The Nematostella genome has a unique set of Adhesion-like sequences lacking GPS domains. There is considerable evidence showing that the Adhesion family is ancestral to the peptide hormone binding Secretin family of GPCRs. PMID- 21618823 TI - 7TM-Cadherins: developmental roles and future challenges. AB - The 7TM-Cadherins, Celsr/Flamingo/Starry night, represent a unique subgroup of adhesion-GPCRs containing atypical cadherin repeats, capable of homophilic interaction, linked to the archetypal adhesion-GPCR seven-transmembrane domain. Studies in Drosophila provided a first glimpse of their functional properties, most notably in the regulation of planar cell polarity (PCP) and in the formation of neural architecture. Many of the developmental functions identified in flies are conserved in vertebrates with PCP predicted to influence the development of multiple organ systems. Details of the molecular and cellular functions of 7TM Cadherins are slowly emerging but many questions remain unanswered. Here the developmental roles of 7TM-Cadherins are discussed and future challenges in understanding their molecular and cellular roles are explored. PMID- 21618824 TI - Latrophilin signalling in tissue polarity and morphogenesis. AB - Understanding the mechanisms that coordinate the polarity of cells and tissues during embryogenesis and morphogenesis is a fundamental problem in developmental biology. We have recently demonstrated that the putative neurotoxin receptor lat 1 defines a mechanism required for the alignment of cell division planes in the early embryo of the nematode C. elegans. Our analysis suggests that lat-1 is required for the propagation rather than the initial establishment of polarity signals. Similar to the role of the flamingo/CELSR protein family in the control of planar cell polarity, these results implicate an evolutionary conserved subfamily of adhesion-GPCRs in the control of tissue polarity and morphogenesis. PMID- 21618825 TI - GPS proteolytic cleavage of adhesion-GPCRs. AB - The stability and functional diversity of proteins can be greatly modulated by posttranslational modification. Proteolytic cleavage at the GPCR proteolysis site (GPS) has been identified as an intrinsic protein modification process of many adhesion-GPCRs. In recentyears, the conserved cleavage site, molecularmechanism and the potential functional implication of the GPS proteolysis have been gradually unveiled. However, many aspects of this unique cleavage reaction including its regulation, the relationship between the cleaved fragments and the functional pathways mediated by the cleaved receptor subunits, remain unanswered. Further investigation of the GPS proteolytic modification shall shed light on the biology of the adhesion-GPCRs. PMID- 21618826 TI - The latrophilins, "split-personality" receptors. AB - Latrophilin, a neuronal "adhesion-G protein-coupled receptor", is the major brain receptor for alpha-latrotoxin, a black widow spidertoxin which stimulates strong neuronal exocytosis in vertebrates. Latrophilin has an unusual structure consisting of two fragments that are produced by the proteolytic cleavage of the parental molecule and that behave independently in the plasma membrane. On binding an agonist, the fragments reassociate and send an intracellular signal. This signal, transduced by a heterotrimeric G protein, causes release of calcium from intracellular stores and massive release of neurotransmitters. Latrophilin represents a phylogenetically conserved family of receptors, with orthologues found in all animals and up to three homologues present in most chordate species. From mammalian homologues, latrophilins 1 and 3 are expressed in neurons, while latrophilin 2 is ubiquitous. Latrophilin 1 may control synapse maturation and exocytosis, whereas latrophilin 2 may be involved in breast cancer. Latrophilins may play different roles during development and in adult animals: thus, LAT-1 determines cell fate in early embryogenesis in Caenorhabditis elegans and controls neurotransmitter release in adult nematodes. This diversity suggests that the functions of latrophilins may be determined by their interactions with respective ligands. The finding of the ligand of latrophilin 1, the large postsynaptic protein lasso, is the first step in the quest for the physiological functions of latrophilins. PMID- 21618827 TI - Studies on the very large G protein-coupled receptor: from initial discovery to determining its role in sensorineural deafness in higher animals. AB - The very large G protein-coupled receptor 1 (VLGRI), also known as MASS1 or GPR98, is most notable among the family of adhesion-GPCR for its size. Encoded by an 18.9 kb open reading frame, the approximately 700 kDa primary translation product is by far the largest GPCR and additionally, the largest cell surface protein known to date. The large ectodomain of the protein contains several repeated motifs, including some 35 calcium binding, Calx-beta repeats and seven copies of an epitempin repeat thought to be associated with the development of epilepsy. The extreme carboxy-terminus contains a consensus PDZ ligand sequence, suggesting interactions with other cytosolic or cytoskeletal proteins. At least two spontaneous and two targeted mutant mouse lines are currently known. The mutant mice present with sensitivity to audiogenic seizures but also have cochlear defects and significant, progressive hearing impairment. Although its ligand is currently unknown, VLGR1 is one of the few adhesion-GPCR family members in which mutations have been shown to be responsible for a human malady. Mutations in VLGRI in humans result in one form (2C) of Usher syndrome, the most common genetic cause of combined blindness and deafness. PMID- 21618828 TI - Adhesion-GPCRs in the CNS. AB - There are a total of 33 members of adhesion G protein-coupled receptors (GPCRs) in humans and 30 members in mice and rats. More than half of these receptors are expressed in the central nervous system (CNS), indicating their possible roles in the development and function of the CNS. Indeed, it has been shown that adhesion GPCRs are involved in the regulation of neurulation, cortical development and neurite growth. Among the few adhesion-GPCRs being studied, GPR56 is so far the only member associated with a human brain malformation called bilateral frontoparietal polymicrogyria (BFPP). The histopathology of BFPP is a cobblestone like brain malformation characterized by neuronal overmigration through a breached pial basement membrane (BM). Further studies in the Gpr56 knockout mouse model revealed that GPR56 is expressed in radial glial cells and regulates the integrity of the pial BM by binding a putative ligand in the extracellular matrix of the developing brain. PMID- 21618829 TI - GPR56 interacts with extracellular matrix and regulates cancer progression. AB - GPR56 is a relatively recent addition to the adhesion-GPCR family. Genetic and biochemical studies uncovered its roles in cancer and development and established its function as an adhesion receptor to mediate the interactions between cells and extracellular matrix. Despite of much progress on understanding its biological implications, the mechanism of its function remains elusive. It has not been firmly established whether GPR56 signals directly through G proteins and what its upstream stimuli and downstream effectors are to execute its various biological effects. This chapter will give an overview of the primary structures of the Gpr56 gene and its encoded protein and attempt to point out open questions in this research area, with an emphasis on its roles in cancer and signal transduction. PMID- 21618830 TI - Adhesion-GPCRS in tumorigenesis. AB - Tumor growth is a highly complex, multistep process that involves tumor cell detachment, migration, invasion and metastasis accompanied by angiogenesis and extracellular matrix turn-over. Each of the steps is influenced by tumor cell interaction and interaction of the tumor cell with its microenvironment that consists of different cell types as tumor-associated fibroblasts, endothelial cells and leukocytes as well as the extracellular matrix produced by the tumor cells themselves or by the fibroblasts. Cellular communication takes place by the regulated expression of adhesion receptors. Adhesion-GPCRs are characterized by very long extracellular N-termini that have multiple domains. When considering this complex structure it is only logical that adhesion-GPCRs are involved in tumor cell interactions. Moreover, these receptors function in cell guidance and/or trafficking, which, in addition to their structure, makes them interesting for tumorigenesis. The aberrant expression of several adhesion-GPCRs on tumor cells and their involvement in tumor growth have been shown for some of the family members. This overview summarizes expression database data as well as data from original research articles of adhesion-GPCRs in tumors. PMID- 21618831 TI - Immunity and adhesion-GPCRs. AB - Adhesion-GPCRs are unusual, owing to their unique structure, comprising a large and complex extracellular domain composed of various common protein modules. Adhesion-GPCR family members are expressed ubiquitously; however the expression of each receptor is highly regulated and often restricted to specific cell types. The EGF-TM7 adhesion-GPCR subfamily members are predominantly expressed by leukocytes and involved in coordinating both the innate and acquired immune responses. Here we highlight some immunological insights in relation to EGF-TM7 proteins and other members of the adhesion-GPCR family. PMID- 21618832 TI - CD97 in leukocyte trafficking. AB - CD97 is a member of the EGF-TM7 family of adhesion G protein-coupled receptors (GPCRs) broadly expressed on leukocytes. CD97 interacts with several cellular ligands via its N-terminal epidermal growth factor (EGF)-like domains. To understand the biological function of CD97, monoclonal antibodies (mAbs) specific for individual EGF domains have been applied in a variety of in vivo models in mice, which represent different aspects of innate and adaptive immunity. Targeting CD97 by mAbs inhibited the accumulation of neutrophilic granulocytes at sites of inflammation thereby affecting antibacterial host defense, inflammatory disorders and stem cell mobilization from bone marrow. Interestingly, targeting CD97 did not impact antigen-specific (adaptive response) models such as delayed type hypersensitivity (DTH) or experimental autoimmune encephalomyelitis (EAE). However, collagen-induced arthritis (CIA), a model for rheumatoid arthritis, was significantly ameliorated suggesting therapeutic value of CD97 targeting. CD97 deficient mice are essentially normal at steady state except for a mild granulocytosis, which increases under inflammatory conditions. Comparison of the consequences of antibody treatment and gene targeting implies that CD97 mAbs actively inhibit the innate response presumably at the level of granulocyte or macrophage recruitment to sites of inflammation. Based on the collected data, we propose that the CD97 mAbs either activate CD97-mediated signal transduction via a yet unknown mechanism or act by inducing CD97 internalization, making CD97 unavailable for binding to its ligands and thereby blocking recruitment of neutrophils and possibly macrophages. PMID- 21618833 TI - The role of CD97 in regulating adaptive T-cell responses. AB - CD97 was identified as an early activation marker on T cells, having low expression on naive T cells. This is a common feature of molecules that have a role in T-cell function. It was subsequently identified as a ligand for CD55, which has been previously identified as an innate regulator of complement. The interaction of this receptor-ligand pair has been shown to provide a potent costimulatory signal to human T cells, despite their modest affinity. Though both CD97 and CD55 are expressed on T cells as well as antigen presenting cells (APCs), their interaction is significant when CD97 on APCs interacts with CD55 on T cells. The converse interaction is poorly defined and may be less significant. A unique aspect of the interaction of CD97 with CD55 is the stimulation of naive T cells, leading to the induction of IL-10 producing cells that behave like Trl regulatory cells. This raises a number of questions regarding the dual functions of CD55; regulating complement and stimulating T cells via CD97 interaction and any potential overlap in the consequences of these dual roles. PMID- 21618834 TI - F4/80: the macrophage-specific adhesion-GPCR and its role in immunoregulation. AB - As a macrophage-restricted reagent, the generation and application of the F4/80 mAb has greatly benefited the phenotypic characterization of mouse tissue macrophages for three decades. Following the molecular identification of the F4/80 antigen as an EGF-TM7 member of the adhesion-GPCR family, great interest was ignited to understand its cell type-specific expression pattern as well as its functional role in macrophage biology. Recent studies have shown that the F4/80 gene is regulated by a novel set of transcription factors that recognized a unique promoter sequence. Gene targeting experiments have produced two F4/80 knock out animal models and showed that F4/80 is not required for normal macrophage development. Nevertheless, the F4/80 receptor was found to be necessary for the induction of efferent CD8+ regulatory T cells responsible for peripheral immune tolerance. The identification of cellular ligands for F4/80 and delineation of its signaling pathway remain elusive but are critical to understand the in vivo role of this macrophage-specific adhesion-GPCR. PMID- 21618835 TI - Signal transduction mediated through adhesion-GPCRs. AB - The signaling cascade of most adhesion-GPCRs remains uncharacterized, as the majority are still orphan receptors and further complicated by their unique structure containing a cleaved long extracellular domain (ECD) and a seven transmembrane domain (7TM). In this chapter, we review previous reports which suggest G protein-dependent and -independent signaling pathways of adhesion-GPCRs and present our approach to investigate the signal transduction of the adhesion GPCR, GPR56. PMID- 21618836 TI - Emerging roles of brain-specific angiogenesis inhibitor 1. AB - Brain-specific angiogenesis inhibitor 1 (BAI1) encodes a seven-transmembrane protein that belongs to the adhesion-GPCR family. Although BAI1 was named for the ability of its extracellular region to inhibit angiogenesis in tumor models, its function in physiological contexts was elusive and remained an orphan receptor until recently. BAI1 is now considered a phagocytic receptor that can recognize phosphatidylserine exposed on apoptotic cells. Moreover, BAI1 has been shown to function upstream of the signaling module comprised of ELMO/Dock180/Rac proteins, thereby facilitating the cytoskeletal reorganization necessary to mediate the phagocytic clearance of apoptotic cells. Here, we review the phylogeny, structure, associating proteins, as well as the known and proposed functions of BAI1. PMID- 21618837 TI - Adhesion-GPCRs in the male reproductive tract. AB - The male reproductive tract expresses a diverse array of adhesion-GPCRs, many in a highly specific and regulated manner. Despite this specificity of expression, little is known about the function of this receptor family in male reproductive physiology. Insights into function are beginning to emerge with the increasing availability of genetically modified mice harbouring mutations in these genes. Gpr64 is the best characterised of the adhesion-GPCRs in the male reproductive system and the phenotype of Gpr64 knock-out mice implicates this receptor in the regulation of fluid absorption in the efferent ducts and proximal epididymis. This chapter summarizes recent data concerning this receptor and other family members in the male reproductive system. PMID- 21618838 TI - Promoting reproductive health knowledge. PMID- 21618839 TI - Centre for Reproductive Health, 2nd Reproductive Health Conference, Lilongwe, Malawi. PMID- 21618840 TI - Patients' satisfaction with reproductive health services at Gogo Chatinkha Maternity Unit, Queen Elizabeth Central Hospital, Blantyre, Malawi. AB - Patient satisfaction is an individual's state of being content with the care provided in the health system. It is important for reproductive health care providers to get feedback from women regarding satisfaction with reproductive health services. There is a dearth of knowledge about patient satisfaction in Malawi. AIM: The specific objective of the study was to determine the extent to which women are satisfied with the care they receive when they come to deliver at the Queen Elizabeth Central Hospital maternity unit. METHODS: A cross sectional study of postpartum women using interviewer administered semi-structured questionnaires was conducted between November 2008 and May 2009. The questionnaires captured mainly quantitative data. RESULTS: 1562 women were interviewed. Most women were housewives (79%) who were referred from Health Centres within the city. Ninety five percent delivered a live baby. The majority of women (97.3%) were satisfied with the care they received from admission through labour and delivery and the immediate postpartum period. Most women cited doctors' and nurses' reviews (65%) as what they liked most about the care they received during their stay in the unit. Most women expected to receive efficient and definitive care. The women's knowledge on patient's rights was extremely low (16%) and equally very few women were offered an opportunity to give an opinion regarding their care by the doctors and nurses in the maternity unit. CONCLUSION: Most women who deliver at the hospital are satisfied with the care offered. This satisfaction is mainly due to the frequent reviews of patients by nurses and doctors in the unit. There is a great need to educate both the population of women served and the health workers that serve them on patient's rights. PMID- 21618841 TI - Empowering young sex workers for safer sex in Dowa and Lilongwe Districts of Malawi. PMID- 21618842 TI - Sentinel surveillance of lymphatic filariasis, schistosomiasis soil transmitted helminths and malaria in rural southern Malawi. AB - BACKGROUND: Baseline prevalence and knowledge, attitude and perception (KAP) survey is a prerequisite for mass drug administration for the control of Lymphatic filariasis (LF) and other neglected tropical diseases. METHODS: In preparation for the first mass drug administration for LF elimination, a baseline survey was conducted in six sentinel sites in the southern Malawi, amongst participants aged five years or more. A standard questionnaire was used to obtain data on socio-demographic factors, ownership and use of bed nets, previous ingestion of ivermectin and KAP toward hydrocele and lymphoedema. Finger prick blood samples were collected from 22:00 to 01:00 hours for LF microscopy, malaria and haemoglobin examination. Stool and urine samples were collected for internal helminths and schistosomiasis respectively. RESULTS: A total of 1, 903 participants were enrolled. Knowledge on the cause of hydrocele and lymphoedema was low in all the sentinel sites (16%-42%, 10%-24% (respectively). Sexual intercourse with a menstruating woman, bad weather and HIV/AIDS were perceived causes of hydrocele. Microfilaraemia prevalence was 1.5% and varied little between sentinel sites (1.0%-2.1%). Childhood urinary schistosomiasis was common in Phalombe (94.9%) and Blantyre (26.9%). CONCLUSION: Integrated approach and understanding of the community KAP is vital or successful implementation of LF elimination programme. PMID- 21618843 TI - Post exposure prophylaxis of HIV transmission after occupational injuries in Queen Elizabeth Central Hospital, Blantyre, Malawi, 2003 - 2008. AB - Health care worker (HCW) in Malawi may acquire HIV infection through occupational injuries, in particular since HIV prevalence among inpatients and incidence of occupational injuries are high. A post exposure prophylaxis (PEP) programme for occupational injuries at Queen Elizabeth Central Hospital (QECH) commenced in 2003. We performed an audit of this programme from 2003 through 2008. 203 Occupational injuries were reported. The majority were needle stick injuries (76.3%). Half of the clients were in a training position. A dual ART regimen was most frequently prescribed. Triple therapy use increased over time and was more frequent in expatriate students. Many nurses and clinical officers were not fully vaccinated for HBV. Based on previous incidence data, occupational injuries were likely to be underreported. Data on side effects were incomplete, however PEP discontinuation due to side effects occurred only twice. Follow up visits were poorly attended, therefore the efficacy of PEP could not be evaluated. Prevention efforts for occupational injuries should be increased and specifically target HCWs in training positions. Measures to improve quality of the PEP programme include effective publicity campaigns, compulsory Hepatitis B vaccination and active tracing of HCWs who default follow up after PEP. PMID- 21618844 TI - Factors involved in selection of a career in surgery and orthopedics for medical students in Malawi. AB - AIMS: There is a critical shortage of Orthopedic Surgeons in Malawi as well as all countries in sub-Saharan Africa. To date, there is no published literature that has investigated surgical or Orthopedic career selection amongst African medical trainees. With the goal of facilitating recruitment into Surgery and Orthopedics in Malawi, we explored the key aspects of Malawian Medical Students' choice of careers in surgical disciplines. METHODS: An on-line survey of all students in clinical years at the College of Medicine in Blantyre, Malawi was performed. The survey was anonymous and constructed de novo by a stringent process including Item Generation, Item reduction, Survey composition, Pre testing, Assessment of Validity by a recognized survey expert, Pilot testing in on-line format by several Malawian Medical Students, and then formal survey testing. RESULTS: Surgery was the most popular specialty choice among the medical trainees (46%). General Surgery was the popular surgical specialty (27%), followed by Neurosurgery (22%) and Orthopedics (19%). The majority of students (67%) feared occupational exposure to HIV but this did not appear to be a factor in specialty choice (p = 0.9). Students with Orthopedic mentors were significantly more likely to choose Orthopedics as their first choice surgical specialty (p = 0.01). Despite limited resources and surgeons in sub-Saharan Africa, surgical specialties are desirable career choices. CONCLUSIONS: This is the first evaluation of factors involved in surgical or Orthopedic career selection in any African context. Future initiatives to improve exposure and mentorship in Orthopedics are fundamental to recruitment into the specialty. PMID- 21618845 TI - Stroke in Malawi--what do we know about it and how should we manage it? Manage it? PMID- 21618846 TI - Ward round--cough, painful throat and progressive hoarseness of voice for 1 year. PMID- 21618847 TI - Connective tissue disease in a Malawian man. PMID- 21618848 TI - The current state of type B aortic dissection. AB - Despite an unprecedented number of publications on all aspects of type B aortic dissections, it remains a formidable disease with barely changed long-term survival over the last decade. In this review, the bewildering amount of information is summarized to distill some practical recommendations for the practitioner faced with one of these patients in the era of endovascular approaches to the aorta. PMID- 21618849 TI - Controversies around carotid stenting. AB - In the last 10 years, CAS became an important alternative to surgery. However, many of the trials regarding stroke prevention had controversial requirements regarding the endovascular approach, leading to questionable conclusions. This review critically addresses results from previous randomized studies on efficacy and safety of the first established therapy for carotid artery stenosis, carotid endarterectomy (CEA) compared to the challenger, carotid artery stenting (CAS), as a newer and less invasive endovascular technique. PMID- 21618850 TI - Initial results after implementation of a multimodal treatment for peritoneal malignancies. AB - INTRODUCTION: Peritoneal carcinomatosis represents a clinical condition with a limited perspective concerning long term survival. The combination of surgical cytoreduction and hyperthermic intraperitoneal chemotherapy (HIPEC) represents a complex multimodal therapeutic management concept with promising results for prolongation of survival. For the identification of pitfalls during implementation of the HIPEC procedure into clinical practice an observational study was conducted. METHODS: Between 2005 and 2009 data from all patients treated with cytoreductive surgery and HIPEC for peritoneal carcinomatosis was prospectively collected and analysed. RESULTS: During the observational interval a total of 42 patients underwent surgical treatment for peritoneal carcinomatosis. In 34 patients the complete procedure with surgical cytoreduction and HIPEC was performed. Perioperative mortality (6%) and morbidity (35%) was similar to other reported series. Twenty-five patients (76%) survived the 18 months follow-up period after complete procedure. CONCLUSION: The multimodal therapeutic treatment concept of surgical cytoreduction and following HIPEC leads to promising results for patients suffering from peritoneal carcinomatosis. However this treatment concept is afflicted with a relevant risk of postoperative complications. PMID- 21618851 TI - The utility of contrast enhanced MR angiography as a first stage diagnostic modality for treatment planning in lower extremity arterial occlusive disease. AB - INTRODUCTION: The aim of this study was to evaluate the applicability of contrast enhanced magnetic resonance angiography (ce-MRA) as a first stage imaging tool for individual treatment planning in patients with lower extremity arterial occlusive disease. PATIENTS AND METHODS: Between August, 2003 and June, 2004, in 128 consecutive patients (182 extremities) with clinical manifestations of lower limb ischemia eligible for invasive therapy, treatment was planned based on clinical assessment, ankle/brachial pressure index measurements combined with ce MRA. Additional duplex ultrasonography (DUS) or digital subtraction angiography (DSA) was done when necessary. Ce-MRA findings were compared with findings during open surgical, endovascular or combined procedures. RESULTS: In 28 extremities (15%) ce-MRA was found inconclusive and additional imaging was performed. In the remaining patients (85% of the extremities (n = 154), treatment was initiated as planned. However, in 19 (11%) of these patients, the treatment plan was altered. In 7 of them, procedural findings did not correspond with those at the time of ce MRA, including 6 patients (3%) with a falsely diagnosed stenosis or occlusion. In total, 62 patients received non-operative treatment (34%), 65 an endovascular procedure (36%), 49 open surgical reconstruction (27%) and 6 a combined treatment. CONCLUSIONS: We conclude that in the majority of patients treatment can be planned based on ce-MRA images, although sometimes additional DUS or DSA may be required. PMID- 21618852 TI - Outcome of patients with ruptured abdominal aortic aneurysm after cardiopulmonary resuscitation. AB - PURPOSE: Ruptured aneurysm of the abdominal aorta (RAAA) is a condition associated with high mortality rate. If Cardiopulmonary Resuscitation (CPR) is required, outcome is considered even worse. The aim of this study was to assess the effect of CPR on 30-day mortality of RAAA patients. Furthermore the Hardman index was evaluated. METHODS: 109 patients with RAAA during a 5 year period (2001 2005) were analysed retrospectively. 30-day mortality, the presence of CPR and Hardman risk factors were recorded. The presence of CPR and the Hardman index were related to clinical outcome. RESULTS: 104 patients were included in our analysis. Eighteen patients received CPR. Overall 30-day mortality was 40%. Patients receiving CPR had a higher mortality rate than patients who did not (89% vs. 30%, p <0.0001). Patients receiving CPR prior to surgery had a mortality rate of 100% (n = 12). In patients with a Hardman Index of < or = 1, 2 and > or = 3 the 30-day mortality was respectively 15%, 47% and 81%. CONCLUSION: Requirement of CPR has a detrimental effect on RAAA-patient outcome. Patients receiving CPR prior to surgery have no survival chance. We advocate that surgery in these patients should not be undertaken. Hardman Index has a predictive value concerning 30-day mortality. PMID- 21618853 TI - Feasibility of a consultant-led, trainee-delivered urgent laparoscopic cholecystectomy service. AB - BACKGROUND: Urgent laparoscopic cholecystectomy has become the gold standard for the treatment of acute gallstone disease. Since 2005 we have implemented a consultant-delivered urgent surgical service for this condition. In an attempt to increase the capacity of this service, we have recently introduced a new policy of also allowing selected trainee surgeons to perform urgent laparoscopic cholecystectomy with consultant assistance available on request. The purpose of this study was to audit our initial experience of this new service. METHODS: Patients with acute gallstone disease had their surgery performed by a consultant or a trainee operating independently with consultant assistance available only on request. Allocation was based purely on surgeon availability. The clinical outcomes of 50 consecutive trainee and 50 consecutive consultant cases were compared and an attempt made to identify pre-operative predictors of technically demanding trainee cases requiring consultant intervention. RESULTS: The mean operating time of trainees was significantly longer than consultants (80 +/- 5 mins vs 55 +/- 4 mins, p <0.001) although the conversion rates for trainees (4%) and consultants (2%) were similar. There were no significant differences between the groups with respect to postoperative morbidity. Of the 50 trainee cases, consultant intervention was required in 12 (24%) cases. There were no statistically significant pre-operative predictors of requirement for consultant assistance. CONCLUSIONS: Urgent laparoscopic cholecystectomy may be performed independently by appropriately skilled trainees within a consultant-led service. Although consultant intervention is often not required, the requirement for consultant assistance cannot be easily predicted based on pre-operative data. PMID- 21618854 TI - Outcome of laparoscopy-assisted feeding tube insertion in elderly stroke victims. AB - BACKGROUND: When percutaneous endoscopic gastrostomy (PEG) is not possible, or fails, the patient is referred for laparoscopic gastrostomy or jejunostomy (LAG/J). METHOD: During 2005-2008, we performed laparoscopy-assisted feeding tube insertion on 15 patients. We assessed the outcome in terms of leaks, infection, longevity etc. The patients were followed-up for up to 12 months. RESULTS: The procedure was successful in 14/15 patients. However, thirteen died within little more than a year, of whom seven suffered from pneumonia or aspiration. DISCUSSION: Our study confirms that LAG/J is technically possible, carries a high morbidity, is a last ditch attempt and that it does not change the general prognosis of these debilitated patients. PMID- 21618855 TI - Giant adrenal myelolipoma: a case report. AB - Adrenal myelolipomas are rare, benign, hormonally inactive tumours composed of mature adipose tissue and haematopoietic elements. Currently, most diagnosed tumours are discovered incidentally because of modern imaging. Myelolipomas are usually asymptomatic, but symptoms such as abdominal pain, haematuria and abdominal mass are described as the result of tumour bulk, haemorrhage or tumour necrosis. Myelolipomas are usually small, although there are descriptions of giant myelolipomas in the literature. We report the case of a giant adrenal myelolipoma in a 79-year-old female who presented with epigastric pain and discomfort. The resected tumour weighed 1777 g and measured 20.5 x 18 x 9.0 cm. PMID- 21618856 TI - A progesterone-receptor-positive huge retroperitoneal tumour mimics metastasis in a breast cancer patient: sarcomatoid renal cell carcinoma. AB - We report a rare case of breast cancer concomitant with progesterone-receptor positive renal cell carcinoma. A 48-year-old woman was diagnosed as having infiltrating ductal carcinoma of the breast and underwent modified radical mastectomy. A synchronous retroperitoneal tumour was detected by sonography of the abdomen in a routine cancer staging. Initially, the tumour was diagnosed as a synchronous retroperitoneal metastasis by needle biopsy; further tests revealed that it was progesterone receptor-positive. The retroperitoneal tumour showed poor response to full courses of adjuvant chemotherapy for breast cancer. Subsequently, the patient underwent a radical operation that included nephrectomy. The final pathology confirmed a sarcomatoid renal cell carcinoma. The post-operative course was uneventful. The patient had no recurrence at the 1 year follow-up. In this report, accurate diagnosis and adequate treatment were discussed. An intra-abdominal tumour with progesterone receptor- (PR) positive features is usually considered to be metastatic in breast cancer patients. For breast cancer patients with a PR-positive retroperitoneal tumour, renal cell carcinoma should be differentiated from a metastatic lesion of breast cancer, even if PR-expression is rare in renal cell carcinoma. To the best of our knowledge, this is the first case of PR-positive expression in breast cancer concomitant with renal carcinoma. In clinical settings, it is challenging for the surgeon to make an accurate diagnosis and to provide prompt treatment in such cases. PMID- 21618857 TI - Mycotic aneurysm of the extracranial carotid artery: a case report and review of the literature. AB - Aneurysm of the extracranial carotid artery is a rare condition; mycotic aneurysms are even less common. They always need surgical treatment to prevent possible rupture, embolisation or death. The literature before 1980 mentions ligation as the only treatment for mycotic aneurysms of the carotid artery, obviously with a high morbidity and mortality rate. We describe an 85-year-old male with a left carotid artery mycotic aneurysm. He presented with a history of Transient Ischaemic Attack (TIA) four weeks after a gastrointestinal infection. Resection of the aneurysm and interposition with autologous vein was performed. Escherichia coli was isolated from the excised tissue. Primary resection of the aneurysm with autologous vein interposition, in association with prolonged antibiotic therapy, is the preferred strategy. Prompt diagnosis and aggressive treatment is essential to prevent serious complications. PMID- 21618858 TI - Necrotizing fasciitis of the abdominal wall following an emergency colostomy: a case report. AB - Necrotizing soft-tissue infections (NSTIs) are acute surgical conditions that demand prompt and multi-faceted treatment. Early recognition, appropriate resuscitation measures, aggressive surgical debridement, and targeted antimicrobial therapy significantly affect the overall outcome and survival of NSTI patients. Necrotizing fasciitis refers to a particular type of NSTI, where necrosis of the skin, subcutaneous tissue and fascia is predominant and there is very little muscle involvement. A 51-year-old woman presented with necrotizing fasciitis of the abdominal wall following colostomy for obstructive colon carcinoma. In this particular case, stoma relocation was necessary because of the need for large parietal surgical debridement. PMID- 21618859 TI - Necrotizing fasciitis of the lower limb after venous surgery: cases studies and a review of the literature. AB - We report 2 cases of necrotizing fasciitis following stripping of the long saphenous vein and phlebectomy of varicose collateral vessels. The first one concerns a 42-year-old man who presented with a left thigh postoperative infection, evolving despite oral antibiotic therapy. Urgent surgical exploration proved an extensive necrosis consistent with necrotizing fasciitis. Wide excision of the necrotic tissue was performed. Under intravenous antibiotic therapy, local wound care and hyperbaric oxygen therapy, the patient's condition improved. The second case concerns a 60-year-old man with cardio-vascular disease and diabetes. He was transferred in our institution 7 days after surgery for an infection in the right thigh and septic shock. Immediate surgical exploration showed extensive necrotizing fasciitis of the thigh, popliteal fossa and latero-posterior compartments of the leg. Muscle necrosis of the right leg was also observed. A right supra-condylar amputation was performed. The patient improved under antibiotherapy and hyperbaric oxygen therapy. PMID- 21618860 TI - Thoughts on a neglected French medieval surgeon: Henri de Mondeville (+/-1260 1320). AB - Henri de Mondeville's manuscript, presented in this article, is based on an incomplete Latin manuscript, written from 1306 to 1320, translated into French by E. Nicaise and kept in the Manuscript section of the Bibliothbque Nationale de Paris (MS French 2030). PMID- 21618861 TI - Uniform talc delivery during video-assisted thoracoscopic pleurodesis. AB - Video-assisted thoracoscopic talc pleurodesis is a highly effective technique to manage recurrent pneumothorax or pleural effusion. However, inhomogenous talc delivery might be a cause of failure. We report a simple and inexpensive technique for uniform talc dispersion with complete covering of the pleural space during video-assisted thoracoscopic pleurodesis. PMID- 21618862 TI - Reliability of computerized cephalometric outcome predictions of mandibular set back surgery. AB - INTRODUCTION: A successful treatment outcome in dentofacial deformity patients commonly requires combined orthodontic-surgical therapy. This enables us to overcome functional, aesthetic and psychological problems. Since most patients state aesthetics as the primary motive for seeking therapy, cephalometric predictions of treatment outcome have become the essential part of treatment planning, especially in combined orthodontic-surgical cases. OBJECTIVE: The aim of this study was to evaluate the validity and reliability of computerized orthognathic surgery outcome predictions generated using the Nemotec Dental Studio NX 2005 software. METHODS: The sample of the study consisted of 31 patients diagnosed with mandibular prognathism who were surgically treated at the Hospital for Maxillofacial Surgery in Belgrade. Investigation was done on lateral cephalograms made before and after surgical treatment. Cephalograms were digitized and analyzed using computer software. According to measurements made on superimposed pre- and postsurgical cephalograms, the patients were retreated within the software and the predictions were assessed by measuring seven angular and three linear parameters. Prediction measurements were then compared with the actual outcome. RESULTS: Results showed statistically significant changes between posttreatment and predicted values for parameters referring to lower lip and mentolabial sulcus position. CONCLUSION: Computerized cephalometric predictions for hard-tissue structures in the sagittal and vertical planes, as well as the VTO parameters, generated using the Nemotec Dental Studio NX 2005 software are reliable, while lower lip and mentolabial sulcus position predictions are not reliable enough. PMID- 21618863 TI - Asthma and chronic bronchitis symptoms among adult population of Belgrade. AB - INTRODUCTION: Over the last three decades the prevalence of respiratory diseases has been increasing worldwide thus increasing economic burden on the healthcare system. Recent studies have shown that the prevalence of asthma in West European countries ranges from 6-9%, while of chronic obstructive pulmonary diseases (COPD) is 8.0% worldwide. OBJECTIVE: The aim of the study was to estimate the prevalence of respiratory symptoms and smoking habits, and to assess the prevalence of asthma and chronic bronchitis among adults in Belgrade, Serbia. METHODS: To collect data we used a questionnaire based on the European Community Respiratory Health Survey (ECRHS) protocol, which was mailed to 10,208 randomly selected subjects. RESULTS: There were 58.3% of responders to our questionnaire. We noted a higher prevalence of respiratory symptoms in subjects who responded promptly. The majority of the respondents were current or former smokers (37.5% and 17.5% respectively) and 79.9% of them reported respiratory symptoms. The most frequent symptoms were longstanding cough (32.2%), sputum production (30.4%) and wheezing (30.3%). Asthma attacks were reported in 4.4% of cases and 5.6% of subjects were using asthma medications. The prevalence of respiratory symptoms increased with age. Women reported coughing, attacks of breathlessness and coughing, chest tightness by night, allergic rhinitis and chronic coughing, more frequently than men. Productive cough was more frequent in men. The prevalence of almost all symptoms was higher in smokers compared to nonsmokers. CONCLUSION: In Serbia there is a high prevalence of respiratory symptoms, asthma and chronic bronchitis smoking addiction. PMID- 21618864 TI - Clinical characteristics of idiopathic ulcerative colitis in children. AB - INTRODUCTION: Idiopathic ulcerative colitis (IUC) represents a rare disease of childhood. It usually occurs at age over 10 years, and below that exceptionally rarely. OBJECTIVE: The aim of the paper was to analyze the clinical signs, symptoms and therapeutic procedures in children with IUC. METHODS: The aims of the paper were based on a sample of 17 children (11 male and 6 female, mean age 11.90 +/- 3.50 years; range 3.8-17.5 years) with IUC. The disease diagnosis was based on characteristic endoscopic and pathohistological findings. RESULTS: The basic signs of the disease involved chronic mucosal haemorrhagic diarrhoea which was confirmed in 16 of 17 patients, with body weight deficiency (10), recurrent abdominal pain (6), fever (5), slowed-down maturation (5), marked anorexia (4), and tenesmus (3).Two patients had recurrent aphthous stomatitis, 2 anal fissures, 2 arthralgia, one autoimmune hepatitis and one pyoderma gangrenosum. None of the children had longitudinal growth retardation. Elevated sedimentation rate and C reactive protein in blood were registered in 11, sideropenia in 10, anaemia in 6 and hypoalbuminemia in 3 patients. The remission of proctosigmoiditis and left sided colitis was achieved with aminosalicylates, and of pancolitis with the combination of aminosalicylates and glucocorticoids, except in cases of steroid dependent colitis, which additionally required azathioprine. CONCLUSION: The main signs of IUC in children are chronic mucous haemorrhagic diarrhoea, body weight loss and sideropenic anaemia, while the basic therapy consists of aminosalicylates, and in more severe cases of the disease the initial use of glucocorticoids and later azathioprine. PMID- 21618865 TI - Henoch-Schonlein purpura outcome in children: a ten-year clinical study. AB - INTRODUCTION: Henoch-Schonlein purpura (HSP) is the most common vasculitis of childhood. It is characterized by symptoms including nonthrombocytopenic purpura, abdominal pain, haematuria/proteinuria, and arthralgia/arthritis. The pleiomorphism of clinical signs in HSP could be confused with other conditions or other vasculitis forms. OBJECTIVE: Evaluation of HSP clinical presentation, the onset and severity of renal manifestation in affected children and their outcome. METHODS: A retrospective study of 49 patients diagnosed with HSP was conducted from September 1999 to September 2009. Children with severe renal manifestations (nephrotic range proteinuria, with or without nephrotic or nephritic syndrome) have undergone kidney biopsy. RESULTS: Twenty-five patients developed renal manifestations after onset of the disease. In our study child's older age was a risk factor for association with HSP nephritis. Six of the patients required kidney biopsy. They were successfully treated with various immunosuppressive protocols, as well as three of nine patients with nephrotic range proteinuria. Two patients developed most severe form of HSP nephritis, nephrotic-nephritic syndrome with histology grade IIIb/IVb. During the study period (average followup 6 years), all patients had a normal global renal function with mild proteinuria in only two cases. The prognosis of renal involvement was better than reports from other patient series. CONCLUSION: Long-term morbidity of HSP is predominantly attributed to renal involvement. During the study period, no patient had renal insufficiency or end stage renal disease after various combinations of immunosuppressive treatment. It is recommended that patients with HSP nephritis are followed for longer periods of time with a regular measurement of renal function and proteinuria. PMID- 21618866 TI - Factors associated with health-related quality of life among university students. AB - INTRODUCTION: University students are subjected to different kinds of stressors, i.e.academic pressures, social issues and financial problems. This can affect their academic achievements and quality of life. OBJECTIVE: The aim of this study was to determine the health-related quality of life of university students, and how it is affected by the presence of chronic diseases, mental disorders comorbidity, and patterns of medical services' use. METHODS: This web-based study included a sample of 1,410 Slovenian university students. We used a self administered questionnaire, containing a sheet with demographic data, Zung's self assessment inventories about anxiety and depression, and EQ-5D questionnaire. The main outcome measures were scores on EQ-5D part and VAS part of the EQ-5D questionnaire. RESULTS: Independent factors associated with the health-related quality of life of university students, were the presence of chronic pain, the presence of depression and anxiety, need for urgent medical help and at least one visit to a clinical specialist in the past year. The independent factors associated with the health status of university students were the presence of chronic diseases, chronic pain, depression and anxiety, a visit to a clinical specialist, a need for urgent medical help and a visit to an emergency unit in the past year. CONCLUSION: Health-related quality of life of university students can be seriously affected by the presence of mental disorders and chronic pain. Appropriate health-related measures should be adopted to achieve early recognition of worse health-related quality of life, the presence of mental disorders and other chronic conditions, and to enable their effective treatment. PMID- 21618867 TI - Acute abdomen caused by adnexal torsion in the first trimester of pregnancy: a case report. AB - INTRODUCTION: Adnexal torsion is a rare cause of acute abdominal pain during pregnancy. The clinical and laboratory findings are non-specific. In this paper we present a case of adnexal torsion in the first trimester of pregnancy. CASE OUTLINE: On admission, the patient presented signs of acute abdomen.The pain started few hours prior to admission, and was predominantly localized, occasionally irradiating to the central parts of the lower abdomen, accompanied by nausea and vomiting. Ultrasound revealed viable intrauterine pregnancy and right adnexal mass with small amount of free fluid in the Douglas pouch. After short preoperative evaluation, laparotomy and adnexectomy were performed. Surgery and postoperative followup were uneventful, and histopathology reported torquated corpus luteum cysts. CONCLUSION: The diagnosis of adnexal torsion during pregnancy is difficult, and occasionally remains a diagnostic dilemma. Surgery is inevitable, must be prompt, and comprises adnexectomy. PMID- 21618868 TI - Chronic fatigue syndrome. AB - Chronic fatigue syndrome (CFS) is defined by a profound, debilitating fatigue, lasting for at least 6 months and resulting in a substantial reduction of occupational, personal, social and educational status. CFS is a relatively poorly recognized clinical entity, although everyday experience shows that there are many patients with CFS symptoms. The incidence and prevalence of CFS remain unknown in most countries; however, the working population is most affected with predominantly female patients in generative period. Although, CFS was first mentioned four centuries ago, mysterious aethiopathogensis of CFS still intrigues scientists as hundreds of studies are still published every year on the subject. About 80 different aetiological CFS factors are mentioned, which can be classified into five basic groups: genetics, immunology, infectious diseases, endocrinology and neuropsychiatry-psychology. Even today the condition is passed established based on the diagnosis by exclusion of organic and psychiatric disorders, which demands a multidisciplinary approach. As the syndrome is often misdiagnosed and mistreated, self-medication is not uncommon in CFS patients'. In addition, such patients usually suffer for years tolerating severe fatigue. Thus, at the moment there are three priorities regarding CFS; understanding pathogenesis, development of diagnostic tests and creating efficient treatment program. PMID- 21618869 TI - RNA exosome. Preface. PMID- 21618870 TI - Finding the exosome. AB - We describe the events surrounding the identification of the exosome complex and the subsequent early development of the field. Like many scientific discoveries, the initial identification and characterization ofthe exosome was based on a combination of skill, good fortune--and the availability of cutting edge technology. PMID- 21618872 TI - The archaeal exosome. AB - The archaeal exosome is aprotein complex with structural similarities to the eukaryotic exosome and bacterial PNPase. Its catalytic core is formed by alternating Rrp41 and Rrp42 polypeptides, arranged in a hexameric ring. A flexible RNA binding cap composed of the evolutionarily conserved proteins Rrp4 and/or Cs14 is bound at the top of the ring and seems to be involved in recruitment of specific substrates and their unwinding. Additionally, the protein complex contains an archaea-specific subunit annotated as DnaG, the function of which is still unknown. The archaeal exosome degrades RNA phosphorolytically in 3' to 5' direction. In a reverse reaction, it synthesizes heteropolymeric RNA tails using nucleoside diphosphates. The functional similarity between the archaeal exosome and PNPase shows that important processes of RNA degradation and posttranscriptional modification in Archaea are similar to the processes in Bacteria and organelles. PMID- 21618871 TI - Structural components and architectures of RNA exosomes. AB - A large body of structural work conducted over the past ten years has elucidated mechanistic details related to 3' to 5' processing and decay of RNA substrates by the RNA exosome. This chapter will focus on the structural organization of eukaryotic exosomes and their evolutionary cousins in bacteria and archaea with an emphasis on mechanistic details related to substrate recognition and to 3' to 5' phosphorolytic exoribonucleolytic activities of bacterial and archaeal exosomes as well as the hydrolytic exoribonucleolytic and endoribonucleolytic activities of eukaryotic exosomes. These points will be addressed in large part through presentation of crystal structures ofphosphorolytic enzymes such as bacterial RNase PH, PNPase and archaeal exosomes and crystal structures ofthe eukaryotic exosome and exosome sub-complexes in addition to standalone structures of proteins that catalyze activities associated with the eukaryotic RNA exosome, namely Rrp44, Rrp6 and their bacterial counterparts. PMID- 21618873 TI - The exosomes of trypanosomes and other protists. AB - The archaeal exosome contains three heterodimeric RNase PH subunits, forming a hexamer with RNase activity; on top sits a trimer of two different SI domain proteins. In animals and yeast, six different, but related subunits form the RNase PH-like core, but these lack enzyme activity; there are three different SI domain proteins and enzyme activity is providedby the endo/exonuclease Rrp44 or- mainly in the nuclear exosome--the Rnase D enzyme Rrp6. Trypanosomes diverged from yeast and mammals very early in eukaryotic evolution. The trypanosome exosome is similar in subunit composition to the human exosome, but instead of being an optional component, trypanosome RRP6 is present in the nucleus and cytoplasm and is required for exosome stability. As in human cells and yeast, the trypanosome exosome has been shown to be required for processing and quality control of rRNA and to be involved in mRNA degradation. Electron microscopy results for a Leishmania exosome suggest that RRP6 is located on the side of the RnasePH ring, interacting with several exosome core proteins. Results of a search for exosome subunits in the genomes of widely diverged protists revealed varied exosome complexity; the Giardia exosome may be as simple as that of Archaea. PMID- 21618874 TI - The exosome and 3'-5' RNA degradation in plants. AB - One of the most versatile RNA degradation machines in eukaryotes is the 3'-5' RNA exosome. It consists of nine conserved subunits forming the core complex, which associates with active ribonucleases, RNA binding proteins, helicases and additional co-factors. While yeast and human exosome core complexes are catalytically inactive, the plant core complex has probably retained a phosphorolytic activity. Intriguingly, the down-regulation of individual subunits of the plant core complex in Arabidopsis mutants led to distinct developmental defects, suggesting an unequal contribution of the core subunits to the in vivo activities of the plant exosome complex. In addition, some of the plant core subunits as well as some associated factors are encoded by duplicated genes, which may have both overlapping and specific functions. Together, these results suggest an unique and complex organisation of exosome-mediated RNA degradation processes in plants. This chapter reviews our current knowledge of plant exosomes and discusses the impact of 3'-5' RNA degradation on the posttranscriptional control of plant genome expression. PMID- 21618875 TI - Catalytic properties of the eukaryotic exosome. AB - The eukaryotic exosome complex is built around the backbone of a 9-subunit ring similar to phosporolytic ribonucleases such as RNase PH and polynucleotide phosphorylase (PNPase). Unlike those enzymes, the ring is devoid of any detectable catalytic activities, with the possible exception of the plant version of the complex. Instead, the essential RNA decay capability is supplied by associated hydrolytic ribonucleases belonging to the Dis3 and Rrp6 families. Dis3 proteins are endowed with two different activities: the long known processive 3' 5' exonucleolytic one and the recently discovered endonucleolytic one. Rrp6 proteins are distributive exonucleases. This chapter will review the current knowledge about the catalytic properties of theses nucleases and their interplay within the exosome holocomplex. PMID- 21618876 TI - Functions of the cytoplasmic exosome. AB - The exosome consists of a core often essential proteins that includes the ribonuclease Rrp44p and is present in both the cytoplasm and nucleus of eukaryotic cells. The cytoplasmic exosome has been extensively characterized in the budding yeast Saccharomyces cerevisiae and some characterization of its metazoan counterpart indicates that most functional aspects are conserved. These studies have implicated the cytoplasmic exosome in the turnover ofnormal cellularmRNAs, as well as several mRNA surveillance pathways. For this, the exosome needs a set of four proteins that do not partake in nuclear exosome functions. These cofactors presumably direct the exosome to specific cytoplasmic RNA substrates. Here, we review cofactors and functions of the cytoplasmic exosome and provide unanswered questions on the mechanisms of cytoplasmic exosome function. PMID- 21618877 TI - Rrp6, Rrp47 and cofactors of the nuclear exosome. AB - This chapter reviews the present state of knowledge on the activity of enzymes that function with the RNA exosome in the nucleus. In this compartment, the exosome interacts physically and functionally with the exoribonuclease Rrp6 and several cofactors, most prominently Rrp47 and the TRAMP complex. These interactions decide the fate of RNA precursors from transcription through the formation of mature ribonucleoprotein particles (RNPs) and the export of the RNPs to the cytoplasm. The nuclear exosome catalyzes the formation of the mature 3' ends of many of these RNAs, but in other cases degrades the RNAs to mononucleotides. Cofactors such as Mpp6, TRAMP and the Nrd1/Nab3 complex play important roles in determining the outcome of the interaction of RNPs with the nuclear exosome. The details that govern the specificity of these decisions remain a rich source for future investigation. PMID- 21618878 TI - The exosome and heterochromatin: multilevel regulation of gene silencing. AB - Heterochromatic silencing is important for repressing gene expression, protecting cells against viral invasion, maintaining DNA integrity and for proper chromosome segregation. Recently, it has become apparent that expression of eukaryotic genomes is far more complex than had previously been anticipated. Strikingly, it has emerged that most of the genome is transcribed including intergenic regions and heterochromatin, calling for us to re-address the question of how gene silencing is regulated and re-evaluate the concept of heterochromatic regions of the genome being transcriptionally inactive. Although heterochromatic silencing can be regulated at the transcriptional level, RNA degrading activities supplied either by the exosome complex or RNAi also significantly contribute to this process. The exosome also regulates noncoding RNAs (ncRNAs) involved in the establishment of heterochromatin, further underscoring its role as the major cellular machinery involved in RNA processing and turn-over. This multilevel control of the transcriptome may be utilized to ensure greater accuracy of gene expression and allow distinction between functional transcripts and background noise. In this chapter, we will discuss the regulation of gene silencing across species, with special emphasis on the exosome's contribution to the process. We will also discuss the links between transcriptional and posttranscriptional mechanisms for gene silencing and their impact on the regulation of eukaryotic transcriptomes. PMID- 21618879 TI - Control of cryptic transcription in eukaryotes. AB - Over the last few years, the development of large-scale technologies has radically modified our conception of genome-wide transcriptional control by unveiling an unexpected high complexity of the eukaryotic transcriptome. In organisms ranging from yeast to human, a considerable number of novel small RNA species have been discovered in regions that were previously thought to be incompatible with high levels of transcription. Intriguingly, these transcripts, which are rapidly targeted for degradation by the exosome, appear to be devoid of any coding potential and may be the consequence of unwanted transcription events. However, the notion that an important fraction of these RNAs representby-products of regulatory transcription is progressively emerging. In this chapter, we discuss the recent advances made in our understanding of the shape of the eukaryotic transcriptome. We also focus on the molecular mechanisms that cells exploit to prevent cryptic transcripts from interfering with the expression of protein-coding genes. Finally, we summarize data obtained in different systems suggesting that such RNAs may play a critical role in the regulation of gene expression as well as the evolution of genomes. PMID- 21618880 TI - The human exosome and disease. AB - Long before the RNA degrading exosome was first described in the yeast Saccharomyces cerevisiae, the use of autoantibodies found in the sera of certain autoimmune patients allowed the identification of a complex of polypeptides which later appeared to be the human exosome. Today, the most extensively documented association of the exosome with disease is still its targeting by the immune system of such patients. The highest frequency of autoantibodies to components of the exosome complex is found in polymyositis-scleroderma overlap patients and therefore the exosome is termed PM/Scl autoantigen in the autoimmune field. More recently, one of the core components of the exosome was identified as a protein associated with chronic myelogenous leukemia. In this chapter we will describe the identification of the PM/Scl autoantigen from a historical perspective, discuss our current knowledge on the occurrence of autoantibodies to exosome components in autoimmune diseases and end with the data that connect the exosome with cancer. PMID- 21618881 TI - Histamine inflammation. Preface. PMID- 21618882 TI - One hundred years of histamine research. AB - In this introductory chapter, we revisit some of the landmarks in the history of histamine research. Since histamine was first synthesized (1907) and isolated as a bacterial contaminant of an extract of ergot (1910), the elucidation of its role in health and disease and its molecular mechanism of action have been continuous, reflecting the application of advances in scientific knowledge, technology and therapeutics over the last 100 years. It appears that the research will continue indefinitely as the nature of the problem is inherently fractal. First, there was a single chemical entity, described in terms of state-of-the art, two-dimensional projections of structures introduced by Fischer in 1891, and an idea that such potent chemicals produced their effects on biological systems as a consequence of an exquisite interaction with a receptive substance, the revolutionary concept of Langley (1905). Today, we recognize four receptor subtypes with multiple activation states and multiple coupling to intracellular effector systems, so that we are no longer able to reliably and in all instances classify compounds interacting with the histamine receptors simply as agonists or antagonists. The complexity is potentially overwhelming, but the promise of value to patients beyond that already provided by the first approved generations of histamine receptor blockers is a compelling driver. PMID- 21618883 TI - Molecular pharmacology of the four histamine receptors. AB - Histamine and its receptors have been (and are still today) very fruitful topics for pharmacological and medicinal chemistry studies. In this chapter we review the various selective ligands that are available for the four different histamine receptors and we describe the main molecular pharmacological aspects of each of the receptor subtypes. PMID- 21618884 TI - Histamine synthesis and lessons learned from histidine decarboxylase deficient mice. AB - This chapter summarizes the information about the transcriptional regulation of histidine decarboxylase (HDC), which is the catabolic enzyme of histamine synthesis, and the activity of histamine in vivo as clarified using HDC gene deficient mice (HDC-KO). The research of the regulatory mechanism of histamine synthesis has been focused on transcriptional and posttranslational aspects. The generation ofHDC-KO mice clarified several new pathophysiological functions of histamine. It is now recognized that the activity of histamine is not limited to allergic, peptic and neurological functions as in the old paradigm, but extends to other fields such as cardiology, immunology and infectious diseases. Therefore, this chapter will focus on these newly revealed functions of histamine. For example, histamine was known to be involved in the effector phase of allergic responses, but a role has now been shown in the sensitization phases and in innate immunity. In the allergic bronchial asthma model using HDC-KO mice it was found that histamine positively controls eosinophilia, but not bronchial hypersensitivity. The effect on eosinophils was afterwards shown to be mediated through the activity of the histamine H4 receptor. The recent advances in the understanding of histamine synthesis and the activity of HDC have dramatically expanded our understanding of the scope of histamine function. PMID- 21618885 TI - Histamine in allergic rhinitis. AB - Histamine plays a major role in allergic rhinitis. In susceptible individuals, allergen induces nasal mast cell degranulation and the release of histamine into the nasal mucosa. Histamine has been detected after controlled challenges with allergen and, when administered into the nasal cavity, elicits signs and symptoms similar to those elicited by allergen. All four histamine receptors have been demonstrated in the nasal mucosa. The role of the four histamine receptors in the pathophysiology of allergic rhinitis are discussed. PMID- 21618886 TI - The role of histamine in ocular allergy. AB - Ocular allergy is a disorder affecting increasing numbers of individuals worldwide. Among the inflammatory mediators that contribute to ocular allergy, histamine is perhaps the best characterized. This monoamine is released by sensitized mast cells upon exposure to allergen and causes symptoms such as redness and tearing. Histamine may also recruit immune cells that can cause long term damage to ocular surfaces. In this chapter we will discuss the known functions of histamine and histamine receptors in ocular allergy and will describe promising therapies targeting the histamine-signaling pathway. PMID- 21618887 TI - The role of histamine in asthma. AB - Histamine is a ubiquitous inflammatory mediator intimately associated with the pathology of allergy. Traditional antihistamines, targeting the histamine H1 receptor, have failed to demonstrate a significant role for histamine in asthma. Novel immunomodulatory roles for histamine and the discovery of a novel histamine receptor, the histamine H4 receptor, have resulted in a reassessment of its importance in asthma. PMID- 21618888 TI - Antihistamines in the treatment of urticaria. AB - Most urticaria subtypes have a profound impact on patients' quality of life and performance. Effective treatment is thus required in all cases where the avoidance of eliciting factors and the elimination of underlying causes is not feasible. In nearly all subtypes histamine released by mast cells plays apredominant role. For symptomatic relief second generation nonsedatinghistamine H1 receptor (H1R)-antihistamines are, therefore, the first choice. However, double-blind controlled studies have shown that dosages required to achieve complete protection from urticaria symptoms may exceed those recommended for other diseases, e.g., allergic rhinitis. Therefore, the current guidelines suggest increasing the dosage up to fourfold, whereas alternative treatments should be reserved as add-on therapy for unresponsive patients. PMID- 21618889 TI - Histamine and antihistamines in atopic dermatitis. AB - Itching (pruritus) is perhaps the most common symptom associated with inflammatory skin diseases and can be a lead symptom ofextracutaneous disease (e.g., malignancy, infection, metabolic disorders). In atopic dermatitis itching sensations constitute one of the most prominent and distressing features. The most characteristic response to itching is the scratch reflex: a more or less voluntary, often sub-conscious motor activity, to counteract the itch by slightly painful stimuli. The benefit of a short-termed relieve from itching through this scratch reflex though is counteracted by a simultaneous damage of the epidermal layer of the skin which leads to increased transepidermal water loss and drying, which in turn results in a cycle of more itching and more scratching. A wide range of peripheral itch-inducing stimuli generated within or administered to the skin are able to trigger pruritus, one of them being histamine. Based on early experiments, histamine has been suggested to may play a key role in the pathogenesis ofAD. This is reflected by a history for antihistamines in the therapeutic medication of AD patients. Antihistamines are believed to share a common antipruritic effect and therefore are prescribed to the vast majority of AD patient suffering from itch to act alleviating. The level of evidence in support of the benefits of antihistamine treatment, however, is low. To assess the benefit of antihistamines in the treatment of AD in a better way, their mechanisms and specific effects need to be understood more precisely. In particular their precise indication is crucial for successful use. This book chapter will therefore summarize and assess the role of histamine in AD and the efficacy of antihistamines in its treatment based on results of basic research and clinical studies. PMID- 21618890 TI - Histamine, immune cells and autoimmunity. AB - Histamine is one ofthe most versatile biogenic amines with multiple roles during the immune response and in allergic disorders. With four distinct G protein coupled receptors (H1R, HER, H3R and H4R), intracellular histamine binding sites (most likely members of the cytochrome P450 family) as well as a membrane transporter (Organic Cation Transporter; OCT3) expressed in various immunocompetent cells, it can entertain a complex network of interactions. These signaling pathways are expressed differentially, depending on the stage of differentiation or activation of target cells, thus adding a further degree of complexity to the system. For this reason, published data are sometimes conflicting and varying according to the particular cell type or responses analyzed and the experimental approaches used. On the other hand, histamine is generated by several cells during the immune response, not only through release of intracellular stores in mast cells or basophils in response to IgE-dependent or -independent stimuli, but also through neosynthesis catalyzed by histidine decarboxylase (HDC) in a number ofhematopoietic cells that secrete the amine immediately without prior storage. These features enable histamine to tune the fine balance between immunity and tolerance by affecting dendritic cells, immunoregulatory cells, T-cell polarization and cytokine production, making the way for new pharmacological strategies to control immune reactivity during immune disorders, such as autoimmunity. PMID- 21618891 TI - Histamine in neurotransmission and brain diseases. AB - Apart from its central role in the mediation of allergic reactions, gastric acid secretion and inflammation in the periphery, histamine serves an important function as a neurotransitter in the central nervous system. The histaminergic neurons originate from the tuberomamillary nucleus of the posterior hypothalamus and send projections to most parts of the brain. The central histamine system is involved in many brain functions such as arousal, control of pituitary hormone secretion, suppression ofeating and cognitive functions. The effects of neuronal histamine are mediated via G-protein-coupled H1-H4 receptors. The prominent role of histamine as a wake-promoting substance has drawn interest to treat sleep-wake disorders, especially narcolepsy, via modulation of H3 receptor function. Post mortem studies have revealed alterations in histaminergic system in neurological and psychiatric diseases. Brain histamine levels are decreased in Alzheimer's disease patients whereas abnormally high histamine concentrations are found in the brains of Parkinson's disease and schizophrenic patients. Low histamine levels are associated with convulsions and seizures. The release of histamine is altered in response to different types of brain injury: e.g. increased release of histamine in an ischemic brain trauma might have a role in the recovery from neuronal damage. Neuronal histamine is also involved in the pain perception. Drugs that increase brain and spinal histamine concentrations have antinociceptive properties. Histaminergic drugs, most importantly histamine H3 receptors ligands, have shown efficacy in many animal models of the above mentioned disorders. Ongoing clinical trials will reveal the efficacy and safety of these drugs in the treatment of human patients. PMID- 21618892 TI - Histamine in normal and malignant cell proliferation. AB - Histamine is a biogenic amine widely distributed throughout the body. Given the observations that histamine can be induced and made available in an unstored diffusible form in tissues undergoing rapid growth (such as tumors and regenerating liver), it could have a role beyond inflammatory and allergic responses. PMID- 21618893 TI - The future antihistamines: histamine H3 and H4 receptor ligands. AB - The field of histamine research has progressed far from a century ago when the first biological functions of histamine were identified. It is now known that histamine function is mediated by four histamine receptors, which belong to the G protein-coupled receptor family. While antihistamines that target the first two receptors have enjoyed clinical and commercial success, efforts to find new antihistamines against the histamine H3 and H4 receptors are still in the early stages. Here we will review the therapeutic potential of targeting these new histamine receptors. PMID- 21618894 TI - An interview with James L. Reinertsen. Interview by Penny Carver. AB - Dr. Reinertsen reflects on improvements in the past decade in the quality and safety of health care and identifies the greatest challenges now facing health care leaders. PMID- 21618895 TI - An interview with John H. Eichhorn. Interview by Joseph Conigliaro. AB - Anesthesiology, with its development of practice standards, helped create the patient safety movement, states Dr. Eichhorn, and "can continue to be the role model and to lead the way in patient safety for all of health care. PMID- 21618896 TI - The safe tables collaborative: a statewide experience. AB - To achieve significant and sustainable statewide improvements in the safety of health care, the Washington State Hospital Association has launched a series of Safe Table Collaboratives. All 97 community hospitals in the state set ambitious goals, implement the latest medical evidence to improve care, and measure progress. PMID- 21618897 TI - Toward the elimination of catheter-related bloodstream infections in a newborn intensive care unit (NICU). AB - At the Children's Hospital at Providence (Anchorage, Alaska) the newborn intensive care unit used a "web of causation" to analyze catheter-related bloodstream infection events and implement evidence-based best practices to reduce or even eliminate them. PMID- 21618898 TI - Using real-time demand capacity management to improve hospitalwide patient flow. AB - BACKGROUND: The Joint Commission's accreditation standard on managing patient flow, effective January 2005, served as a call to action for hospitals, yet many hospitals still lack the processes and structures to admit or transfer patients to an inpatient bed on a timely basis. In 2007 the University of Pittsburgh Medical Center (UPMC) at Shadyside, a 526-bed tertiary care hospital, began testing and implementing real-time demand capacity management (RTDC) at an initial pilot site. The hospital had identified improved patient flow as a strategic goal in 2002, but a series of patient flow projects failed to result in improvement. IMPLEMENTING RTDC: Standard processes for the four RTDC steps Predicting Capacity, Predicting Demand, Developing a Plan, and Evaluating a Plan- and standard structures for unit bed huddles and the hospital bed meetings were developed. The neurosurgery (NS) service line's ICU and stepdown unit were designated as the first pilot sites, but work was quickly spread to other units. RESULTS: Improvements were achieved and have been sustained through early 2011 for all measures, including (1) the unit-based reliability of discharge predictions; (2) overnight holds in the postanesthesia care unit, a problem eliminated two months after RTDC work began; (3) the percentage of patients who left without being seen (LWBS), routinely < 0.5% by May 2008; (5) the emergency department median length of stay for admitted patients, routinely < 4 hours after March 2008; and (6) aggregate length of stay (ALOS), generally maintained at < 5.75 days. CONCLUSIONS: RTDC represents a promising approach to improving hospitalwide patient flow. Its four steps, integrated into current bed management processes, are not an add-on to the work needing to be accomplished everyday. PMID- 21618899 TI - ProvenCare perinatal: a model for delivering evidence/ guideline-based care for perinatal populations. AB - BACKGROUND: Geisinger Health System (GHS) has applied its ProvenCare model to demonstrate that a large integrated health care delivery system, enabled by an electronic health record (EHR), could reengineer a complicated clinical process, reduce unwarranted variation, and provide evidence-based care for patients with a specified clinical condition. In 2007 GHS began to apply the model to a more complicated, longer-term condition of "wellness"--perinatal care. ADAPTING PROVENCARE TO PERINATAL CARE: The ProvenCare Perinatal initiative was more complex than the five previous ProvenCare endeavors in terms of breadth, scope, and duration. Each of the 22 sites created a process flow map to depict the current, real-time process at each location. The local practice site providers physicians and mid-level practitioners-reached consensus on 103 unique best practice measures (BPMs), which would be tracked for every patient. These maps were then used to create a single standardized pathway that included the BPMs but also preserved some unique care offerings that reflected the needs of the local context. RESULTS: A nine-phase methodology, expanded from the previous six-phase model, was implemented on schedule. Pre- to postimplementation improvement occurred for all seven BPMs or BPM bundles that were considered the most clinically relevant, with five statistically significant. In addition, the rate of primary cesarean sections decreased by 32%, and birth trauma remained unchanged as the number of vaginal births increased. CONCLUSIONS: Preliminary experience suggests that integrating evidence/guideline-based best practices into work flows in inpatient and outpatient settings can achieve improvements in daily patient care processes and outcomes. PMID- 21618900 TI - Chronic myelogenous leukemia in the chronic phase with lymph node swelling which represented extramedullary involvement composed of cells at different stages of maturation. AB - In chronic myelogenous leukemia (CML), lymph node swelling is generally considered an early sign of blast crisis. We encountered a rare case of CML showing marked lymph node swelling, which represented extramedullary involvement composed of cells at different stages of maturation but not showing blast crisis. In 1996, a 60-year-old Japanese male was diagnosed with CML in the chronic phase, and imatinib mesylate was prescribed following interferon-alpha treatment in 2004. However, the patient chose to discontinue out patient treatment 1 year later. He was referred to our hospital because of severe fatigue in August 2006, and laboratory tests showed a WBC count of 347.9 x 10(9)/l with 6.5% blasts. On bone marrow tests, Ph chromosome presence was found in all cells examined, but there was no additional chromosomal abnormality. Cytogenetic analysis revealed that 82% of cells in on employing FISH showed BCR/ABL. The patient had swelling of the left cervical and both inguinal lymph nodes. A lymph node biopsy showed proliferation in three lineages of hematological cells at different stages of maturation. He was diagnosed with CML in the chronic phase. He was given imatinib mesylate and hydroxyurea, and achieved hematological remission. Sixteen months later, he developed blast crisis and died of cerebral hemorrhage. Such a case of extramedullary involvement composed of cells at different stages of maturation is rare, and suggests the need to biopsy extramedullary tumors in order to evaluate the clinical phase. PMID- 21618901 TI - Reduced locomotor sensitization induced by methamphetamine and altered gene expression in ICER overexpressing mice. PMID- 21618902 TI - Comparison of the depression-like behavior and serotonergic system between Wistar and Wistar-Kyoto rat strains. PMID- 21618903 TI - Influence of GIRK channel inhibition on alcohol abstinence and relapse risk in Japanese alcohol-dependent outpatients. PMID- 21618904 TI - Effect of chronic lithium treatment on gene expression profile in mouse microglia and brain dendritic cells. PMID- 21618905 TI - Metacommunity assembly and sorting in newly formed lake communities. AB - new habitats are created, community assembly may follow independent trajectories, since the relative importance of dispersal limitation, priority effects, species interactions, and environmental gradients can vary as assembly proceeds. Unfortunately, tracking community colonization and composition across decades is challenging. We compiled a multiyear community composition data set and reconstructed past communities with remains from sediment cores to investigate cladoceran assembly dynamics in six older (1920s) and two more recently formed (1950s) lakes. We found that current communities cluster along a gradient of thermal stratification that is known to influence predation intensity. Assembling communities showed evidence for a greater influence of species sorting and a reduced influence of spatial structure since the first colonizations. However, lake community trajectories varied considerably, reflecting different colonization sequences among lakes. In the older lakes, small-bodied cladocerans often arrived much earlier than large-bodied cladocerans, while the two younger lakes were colonized much more rapidly, and one was quickly dominated by a large bodied species. Thus, by combining contemporary community data with paleoecological records, we show that assembly history influences natural community structure for decades while patterns of ecological sorting develop. PMID- 21618906 TI - Latitudinal variation in top-down and bottom-up control of a salt marsh food web. AB - The shrub Iva frutescens, which occupies the terrestrial border of U.S. Atlantic Coast salt marshes, supports a food web that varies strongly across latitude. We tested whether latitudinal variation in plant quality (higher at high latitudes), consumption by omnivores (a crab, present only at low latitudes), consumption by mesopredators (ladybugs, present at all latitudes), or the life history stage of an herbivorous beetle could explain continental-scale field patterns of herbivore density. In a mesocosm experiment, crabs exerted strong top-down control on herbivorous beetles, ladybugs exerted strong top-down control on aphids, and both predators benefited plants through trophic cascades. Latitude of plant origin had no effect on consumers. Herbivorous beetle density was greater if mesocosms were stocked with beetle adults rather than larvae, and aphid densities were reduced in the "adult beetle" treatment. Treatment combinations representing high and low latitudes produced patterns of herbivore density similar to those in the field. We conclude that latitudinal variation in plant quality is less important than latitudinal variation in top consumers and competition in mediating food web structure. Climate may also play a strong role in structuring high-latitude salt marshes by limiting the number of herbivore generations per growing season and causing high overwintering mortality. PMID- 21618907 TI - Bottom-up control of consumers leads to top-down indirect facilitation of invasive annual herbs in semiarid Chile. AB - The abundance of exotic plants is thought to be limited by competition with resident species (including plants and generalist herbivores). In contrast, observations in semiarid Chile suggest that a native generalist rodent, the degu (Octodon degus), may be facilitating the expansion of exotic annual plants. We tested this hypothesis with a 20-year data set from a World Biosphere Reserve in mediterranean Chile. In this semiarid environment, rainfall varies annually and dramatically influences cover by both native and exotic annual plants; degu population density affects the composition and cover of exotic and native annual plants. In low-rainfall years, cover of both native and exotic herbs is extremely low. Higher levels of precipitation result in proportional increases in cover of all annual plants (exotic and native species), leading in turn to increases in degu population densities, at which point they impact native herbs in proportion to their greater cover, indirectly favoring the expansion of exotic plants. We propose that bottom-up control of consumers at our site results in top-down indirect facilitation of invasive annual herbs, and that this pattern may be general to other semiarid ecosystems. PMID- 21618908 TI - Making more out of sparse data: hierarchical modeling of species communities. AB - Community ecologists and conservation biologists often work with data that are too sparse for achieving reliable inference with species-specific approaches. Here we explore the idea of combining species-specific models into a single hierarchical model. The community component of the model seeks for shared patterns in how the species respond to environmental covariates. We illustrate the modeling framework in the context of logistic regression and presence-absence data, but a similar hierarchical structure could also be used in many other types of applications. We first use simulated data to illustrate that the community component can improve parameterization of species-specific models especially for rare species, for which the data would be too sparse to be informative alone. We then apply the community model to real data on 500 diatom species to show that it has much greater predictive power than a collection of independent species specific models. We use the modeling approach to show that roughly one-third of distance decay in community similarity can be explained by two variables characterizing water quality, rare species typically preferring nutrient-poor waters with high pH, and common species showing a more general pattern of resource use. PMID- 21618909 TI - Soil microbes drive the classic plant diversity-productivity pattern. AB - Ecosystem productivity commonly increases asymptotically with plant species diversity, and determining the mechanisms responsible for this well-known pattern is essential to predict potential changes in ecosystem productivity with ongoing species loss. Previous studies attributed the asymptotic diversity-productivity pattern to plant competition and differential resource use (e.g., niche complementarity). Using an analytical model and a series of experiments, we demonstrate theoretically and empirically that host-specific soil microbes can be major determinants of the diversity-productivity relationship in grasslands. In the presence of soil microbes, plant disease decreased with increasing diversity, and productivity increased nearly 500%, primarily because of the strong effect of density-dependent disease on productivity at low diversity. Correspondingly, disease was higher in plants grown in conspecific-trained soils than heterospecific-trained soils (demonstrating host-specificity), and productivity increased and host-specific disease decreased with increasing community diversity, suggesting that disease was the primary cause of reduced productivity in species-poor treatments. In sterilized, microbe-free soils, the increase in productivity with increasing plant species number was markedly lower than the increase measured in the presence of soil microbes, suggesting that niche complementarity was a weaker determinant of the diversity-productivity relationship. Our results demonstrate that soil microbes play an integral role as determinants of the diversity-productivity relationship. PMID- 21618910 TI - Dangerous liaisons disperse the Mediterranean dwarf palm: fleshy-pulp defensive role against seed predators. AB - We chose the interaction between the Mediterranean dwarf palm (Chamaerops humilis) and its major seed disperser, the Eurasian badger (Meles meles), to evaluate the hypothesis that endozoochory is characterized by a mixture of conflicting and overlapping interests, with the capacity of being positive or negative for plant fitness. For instance, because of the potential protective role against invertebrate seed predators of C. humilis pulp, we expected that badger-ingested diaspores (i.e., seeds without pulp) would show lower survival than control seeds with the pulp attached. Conversely, due to the possible germination inhibitory function of C. humilis fruit pulp, it was also likely that badger-ingested seeds germinate in higher proportion than control seeds. We evaluated our predictions by carrying out a field sowing and monitoring it over two years. We estimated several stage-specific transition probabilities as well as the cumulative probability of seedling recruitment under different treatments. Our experimental results revealed the multifunctionality of C. humilis fruit pulp and that fruit ingestion by badgers had conflicting outcomes for the palm. As predicted, seed survival was much lower and seed predation by invertebrates much higher for badger-ingested than for control seeds, suggesting a defensive role of C. humilis ripe fruit pulp. All early-emerged seedlings came from badger-ingested seeds, suggesting an inhibitory function of fruit pulp. Though we did not find an effect of removal from the maternal environment on most components of fitness, seedling survival for badger-ingested seeds was higher away from than beneath conspecifics. Badgers imposed a sizeable short-term fitness cost to C. humilis and therefore could be categorized as a "dangerous liaison." Nonetheless, because of the high mobility of the badger, its dispersal service appears paramount given the severe fragmentation and isolation of most C. humilis populations across the highly humanized Mediterranean basin. Our study thus illustrates the necessity of assessing concurrently direct and indirect effects of plant-disperser interactions at different stages of the plant life cycle and recruitment process. PMID- 21618911 TI - Volatile chemicals from leaf litter are associated with invasiveness of a neotropical weed in Asia. AB - Some invasive plant species appear to strongly suppress neighbors in their nonnative ranges but much less so in their native range. We found that in the field in its native range in Mexico, the presence of Ageratina adenophora, an aggressive Neotropical invader, was correlated with higher plant species richness than found in surrounding plant communities where this species was absent, suggesting facilitation. However, in two nonnative ranges, China and India, A. adenophora canopies were correlated with much lower species richness than the surrounding communities, suggesting inhibition. Volatile organic compound (VOC) signals may contribute to this striking biogeographical difference and the invasive success of A. adenophora. In controlled experiments volatiles from A. adenophora litter caused higher mortality of species native to India and China, but not of species native to Mexico. The effects of A. adenophora VOCs on seedling germination and growth did not differ between species from the native range and species from the nonnative ranges of the invader. Litter from A. adenophora plants from nonnative populations also produced VOCs that differed quantitatively in the concentrations of some chemicals than litter from native populations, but there were no chemicals unique to one region. Biogeographic differences in the concentrations of some volatile compounds between ranges suggest that A. adenophora may be experiencing selection on biochemical composition in its nonnative ranges. PMID- 21618912 TI - Macronutrient content of plant-based food affects growth of a carnivorous arthropod. AB - Many arthropods engage in mutualisms in which they consume plant-based foods including nectar, extrafloral nectar, and honeydew. However, relatively little is known about the manner in which the specific macronutrients in these plant-based resources affect growth, especially for carnivorous arthropods. Using a combination of laboratory and field experiments, we tested (1) how plant-based foods, together with ad libitum insect prey, affect the growth of a carnivorous ant, Solenopsis invicta, and (2) which macronutrients in these resources (i.e., carbohydrates, amino acids, or both) contribute to higher colony growth. Access to honeydew increased the production of workers and brood in experimental colonies. This growth effect appeared to be due to carbohydrates alone as colonies provided with the carbohydrate component of artificial extrafloral nectar had greater worker and brood production compared to colonies deprived of carbohydrates. Surprisingly, amino acids only had a slight interactive effect on the proportion of a colony composed of brood and negatively affected worker survival. Diet choice in the laboratory and field matched performance in the laboratory with high recruitment to carbohydrate baits and only slight recruitment to amino acids. The strong, positive effects of carbohydrates on colony growth and the low cost of producing this macronutrient for plants and hemipterans may have aided the evolution of food-for-protection mutualisms and help explain why these interactions are so common in ants. In addition, greater access to plant-based resources in the introduced range of S. invicta may help to explain the high densities achieved by this species throughout the southeastern United States. PMID- 21618913 TI - Effects of combining an intraguild predator with a cannibalistic intermediate predator on a species-level trophic cascade. AB - A greater diversity of natural enemies can in some cases disrupt prey suppression, particularly when natural enemies engage in intraguild predation, where natural enemies compete with and prey upon each other. However, empirical studies have often demonstrated enhanced prey suppression despite intraguild predation. A recent theoretical study proposed the hypothesis that, when the intermediate predator is cannibalistic, intraguild predation can reduce cannibalism within the intermediate predator population, leading to little change in intermediate predator mortality and thus enhanced prey suppression. The goal of this study was to examine this hypothesis empirically. Two summer-long field enclosure experiments were conducted in cotton fields. We investigated the effects of adding an intraguild predator, Zelus renardii, on (1) the abundance of a cannibalistic intermediate predator, Geocoris pallens, (2) the abundance of a herbivore, Lygus hesperus, and (3) cotton plant performance. G. pallens adult abundance did not increase, even when food availability was high and natural enemies were absent, suggesting that density-dependent cannibalism imposes an upper limit on its densities. Furthermore, although Z. renardii is an intraguild predator of G. pallens, G. pallens long-term densities were unaffected by Z. renardii. In the presence of the intermediate predator, the addition of the intraguild predator Z. renardii enhanced suppression of L. hesperus, and there were suggestions that Z. renardii and G. pallens partitioned the L. hesperus population. Effects of herbivore suppression cascaded to the plant level, improving plant performance. In conclusion, we provide empirical support for the hypothesis that the addition of an intraguild predator may enhance prey suppression if the intermediate predator expresses density-dependent cannibalism. Intraguild predation and cannibalism co-occur in many communities; thus their joint effects may be broadly important in shaping predator effects on herbivores and plant performance. PMID- 21618914 TI - Monarch butterfly migration and parasite transmission in eastern North America. AB - Seasonal migration occurs in many animal systems and is likely to influence interactions between animals and their parasites. Here, we focus on monarch butterflies (Danaus plexippus) and a protozoan parasite (Ophryocystis elektroscirrha) to investigate how host migration affects infectious disease processes. Previous work showed that parasite prevalence was lower among migratory than nonmigratory monarch populations; two explanations for this pattern are that (1) migration allows animals to periodically escape contaminated habitats (i.e., migratory escape), and (2) long-distance migration weeds out infected animals (i.e., migratory culling). We combined field-sampling and analysis of citizen science data to examine spatiotemporal trends of parasite prevalence and evaluate evidence for these two mechanisms. Analysis of within breeding-season variation in eastern North America showed that parasite prevalence increased from early to late in the breeding season, consistent with the hypothesis of migratory escape. Prevalence was also positively related to monarch breeding activity, as indexed by larval density. Among adult monarchs captured at different points along the east coast fall migratory flyway, parasite prevalence declined as monarchs progressed southward, consistent with the hypothesis of migratory culling. Parasite prevalence was also lower among monarchs sampled at two overwintering sites in Mexico than among monarchs sampled during the summer breeding period. Collectively, these results indicate that seasonal migration can affect parasite transmission in wild animal populations, with implications for predicting disease risks for species with threatened migrations. PMID- 21618915 TI - Intraguild predation in the presence of a shared natural enemy. AB - Intraguild predation (IGP) is a widespread phenomenon in nature, and yet the simplest theoretical models of IGP predict that coexistence of intraguild predator and prey is only possible under restrictive assumptions. Here I examine how a specialist or generalist natural enemy of these species affects their long term persistence and abundance, as functions of the natural enemy's relative attack severity and fecundity on each species. Notably, I show that failure to include the effects of a higher trophic level in models of IGP can lead to incorrect predictions about the coexistence or exclusion of guild members. I then consider how an interaction between native species and a natural enemy is perturbed by the arrival of an invasive intraguild predator. I outline the conditions under which the native species and/or its natural enemy are threatened by the arrival of the intraguild predator, and also when the natural enemy is beneficial in preventing the initial invasion or eventual dominance of the invader. This work provides new insights on the influence of omnivory on food web stability, and also generates testable hypotheses for predicting the impact of a novel intraguild predator on the recipient community at multiple trophic levels. PMID- 21618916 TI - Stochastic growth reduces population fluctuations in Daphnia-algal systems. AB - Deterministic, size-structured models are widely used to describe consumer resource interactions. Such models typically ignore potentially large random variability in juvenile development rates. We present simple representations of this variability and show five approaches to calculating the model parameters for Daphnia pulex interacting with its algal food. Using our parameterized models of growth variability, we investigate the robustness of a recently proposed stabilizing mechanism for Daphnia populations. Growth rate variability increases the range of enrichments over which small-amplitude cycles or quasi-cycles occur, thus increasing the plausibility that the underlying mechanism contributes to the prevalence of small-amplitude cycles in the field and in experiments. More generally, our approach allows us to relate commonly available information on variance of development times to population stability. PMID- 21618917 TI - Marine-derived nutrients, bioturbation, and ecosystem metabolism: reconsidering the role of salmon in streams. AB - In coastal areas of the North Pacific Ocean, annual returns of spawning salmon provide a substantial influx of nutrients and organic matter to streams and are generally believed to enhance the productivity of recipient ecosystems. Loss of this subsidy from areas with diminished salmon runs has been hypothesized to limit ecosystem productivity in juvenile salmon rearing habitats (lakes and streams), thereby reinforcing population declines. Using five to seven years of data from an Alaskan stream supporting moderate salmon densities, we show that salmon predictably increased stream water nutrient concentrations, which were on average 190% (nitrogen) and 390% (phosphorus) pre-salmon values, and that primary producers incorporated some of these nutrients into tissues. However, benthic algal biomass declined by an order of magnitude despite increased nutrients. We also measured changes in stream ecosystem metabolic properties, including gross primary productivity (GPP) and ecosystem respiration (ER), from three salmon streams by analyzing diel measurements of oxygen concentrations and stable isotopic ratios (delta O-O2) within a Bayesian statistical model of oxygen dynamics. Our results do not support a shift toward higher primary productivity with the return of salmon, as is expected from a nutrient fertilization mechanism. Rather, net ecosystem metabolism switched from approximately net autotrophic (GPP > or = ER) to a strongly net heterotrophic state (GPP << ER) in response to bioturbation of benthic habitats by salmon. Following the seasonal arrival of salmon, GPP declined to <12% of pre-salmon rates, while ER increased by over threefold. Metabolism by live salmon could not account for the observed increase in ER early in the salmon run, suggesting salmon nutrients and disturbance enhanced in situ heterotrophic respiration. Salmon also changed the physical properties of the stream, increasing air-water gas exchange by nearly 10 fold during peak spawning. We suggest that management efforts to restore salmon ecosystems should consider effects on ecosystem metabolic properties and how salmon disturbance affects the incorporation of marine-derived nutrients into food webs. PMID- 21618918 TI - Role of the fish Astyanax aeneus (Characidae) as a keystone nutrient recycler in low-nutrient neotropical streams. AB - Nutrient recycling by animals is a potentially important biogeochemical process in both terrestrial and aquatic ecosystems. Stoichiometric traits of individual species may result in some taxa playing disproportionately important roles in the recycling of nutrients relative to their biomass, acting as keystone nutrient recyclers. We examined factors controlling the relative contribution of 12 Neotropical fish species to nutrient recycling in four streams spanning a range of phosphorus (P) levels. In high-P conditions (135 microg/L soluble reactive phosphorus, SRP), most species fed on P-enriched diets and P excretion rates were high across species. In low-P conditions (3 microg/L SRP), aquatic food resources were depleted in P, and species with higher body P content showed low rates of P recycling. However, fishes that were subsidized by terrestrial inputs were decoupled from aquatic P availability and therefore excreted P at disproportionately high rates. One of these species, Astyanax aeneus (Characidae), represented 12% of the total population and 18% of the total biomass of the fish assemblage in our focal low-P study stream but had P excretion rates > 10-fold higher than other abundant fishes. As a result, we estimated that P excretion by A. aeneus accounted for 90% of the P recycled by this fish assemblage and also supplied approximately 90% of the stream P demand in this P-limited ecosystem. Nitrogen excretion rates showed little variation among species, and the contribution of a given species to ecosystem N recycling was largely dependent upon the total biomass of that species. Because of the high variability in P excretion rates among fish species, ecosystem-level P recycling could be particularly sensitive to changes in fish community structure in P limited systems. PMID- 21618919 TI - Minimizing predation risk in a landscape of multiple predators: effects on the spatial distribution of African ungulates. AB - Studies that focus on single predator-prey interactions can be inadequate for understanding antipredator responses in multi-predator systems. Yet there is still a general lack of information about the strategies of prey to minimize predation risk from multiple predators at the landscape level. Here we examined the distribution of seven African ungulate species in the fenced Karongwe Game Reserve (KGR), South Africa, as a function of predation risk from all large carnivore species (lion, leopard, cheetah, African wild dog, and spotted hyena). Using observed kill data, we generated ungulate-specific predictions of relative predation risk and of riskiness of habitats. To determine how ungulates minimize predation risk at the landscape level, we explicitly tested five hypotheses consisting of strategies that reduce the probability of encountering predators, and the probability of being killed. All ungulate species avoided risky habitats, and most selected safer habitats, thus reducing their probability of being killed. To reduce the probability of encountering predators, most of the smaller prey species (impala, warthog, waterbuck, kudu) avoided the space use of all predators, while the larger species (wildebeest, zebra, giraffe) only avoided areas where lion and leopard space use were high. The strength of avoidance for the space use of predators generally did not correspond to the relative predation threat from those predators. Instead, ungulates used a simpler behavioral rule of avoiding the activity areas of sit-and-pursue predators (lion and leopard), but not those of cursorial predators (cheetah and African wild dog). In general, selection and avoidance of habitats was stronger than avoidance of the predator activity areas. We expect similar decision rules to drive the distribution pattern of ungulates in other African savannas and in other multi-predator systems, especially where predators differ in their hunting modes. PMID- 21618920 TI - Using multilevel spatial models to understand salamander site occupancy patterns after wildfire. AB - Studies of the distribution of elusive forest wildlife have suffered from the confounding of true presence with the uncertainty of detection. Occupancy modeling, which incorporates probabilities of species detection conditional on presence, is an emerging approach for reducing observation bias. However, the current likelihood modeling framework is restrictive for handling unexplained sources of variation in the response that may occur when there are dependence structures such as smaller sampling units that are nested within larger sampling units. We used multilevel Bayesian occupancy modeling to handle dependence structures and to partition sources of variation in occupancy of sites by terrestrial salamanders (family Plethodontidae) within and surrounding an earlier wildfire in western Oregon, USA. Comparison of model fit favored a spatial N mixture model that accounted for variation in salamander abundance over models that were based on binary detection/non-detection data. Though catch per unit effort was higher in burned areas than unburned, there was strong support that this pattern was due to a higher probability of capture for individuals in burned plots. Within the burn, the odds of capturing an individual given it was present were 2.06 times the odds outside the burn, reflecting reduced complexity of ground cover in the burn. Ther was weak support that true occupancy was lower within the burned area. While the odds of occupancy in the burn were 0.49 times the odds outside the burn among the five species, the magnitude of variation attributed to the burn was small in comparison to variation attributed to other landscape variables and to unexplained, spatially autocorrelated random variation. While ordinary occupancy models may separate the biological pattern of interest from variation in detection probability when all sources of variation are known, the addition of random effects structures for unexplained sources of variation in occupancy and detection probability may often more appropriately represent levels of uncertainty. PMID- 21618921 TI - The ecological impact of invasive cane toads on tropical snakes: field data do not support laboratory-based predictions. AB - Predicting which species will be affected by an invasive taxon is critical to developing conservation priorities, but this is a difficult task. A previous study on the impact of invasive cane toads (Bufo marinus) on Australian snakes attempted to predict vulnerability a priori based on the assumptions that any snake species that eats frogs, and is vulnerable to toad toxins, may be at risk from the toad invasion. We used time-series analyses to evaluate the accuracy of that prediction, based on >3600 standardized nocturnal surveys over a 138-month period on 12 species of snakes and lizards on a floodplain in the Australian wet dry tropics, bracketing the arrival of cane toads at this site. Contrary to prediction, encounter rates with most species were unaffected by toad arrival, and some taxa predicted to be vulnerable to toads increased rather than declined (e.g., death adder Acanthophis praelongus; Children's python Antaresia childreni). Indirect positive effects of toad invasion (perhaps mediated by toad induced mortality of predatory varanid lizards) and stochastic weather events outweighed effects of toad invasion for most snake species. Our study casts doubt on the ability of a priori desktop studies, or short-term field surveys, to predict or document the ecological impact of invasive species. PMID- 21618922 TI - The effects of opportunistic and intentional predators on the herding behavior of prey. AB - In this article, we study how predator behavior influences the aggregation of prey into herds. Game-theoretic models of herd formation are developed based on different survival probabilities of solitary prey and prey that join the herd and on the predator's preference of what type of prey to search for. For an intentional predator that will only pursue its preferred type of prey, a single herd with no solitaries cannot emerge unless the herd acts as a prey refuge. If neither prey choice provides a refuge, it is shown that an equilibrium always exists where there are both types of prey and the predator does not always search for the same type of prey (i.e., a mixed equilibrium exists). On the other hand, if the predator is opportunistic in that it sometimes shifts to pursue the type of prey that is observed first, there may be a single herd equilibrium that does not act as a prey refuge when there is a high level of opportunistic behavior. For low opportunistic levels, a mixed equilibrium is again the only outcome. The evolutionary stability of each equilibrium is tested to see if it predicts the eventual herding behavior of prey in its corresponding model. Our analysis confirms that both predator and prey preferences (for herd or solitary) have strong effects on why prey aggregate. In particular, in our models, only the opportunistic predator can maintain all prey in a single herd that is under predation risk. PMID- 21618923 TI - Sex ratio bias and reproductive strategies: what sex to produce when? AB - Several theories predict the evolution of bias in progeny sex ratio based on variations in maternal or offspring reproductive value. For mammals, however, tests of sex-bias theories have produced inconsistent results, and no clear patterns have emerged. Each theory is based on assumptions that are difficult to satisfy, and empirical tests require large data sets. Using a long-term study on bighorn sheep (Ovis canadensis), we identified several parameters that influence progeny sex ratio according to maternal state. For older females, progeny sex ratio was affected by an interaction between reproductive strategy and environmental conditions. When conditions were good, old females reproduced every year but minimized fitness costs by producing daughters. When conditions were poor, old females produced more sons but did not reproduce every year. Sons of older females were of similar mass to those born to younger females under poor conditions but were smaller and likely disadvantaged under good environmental conditions. For young and prime-aged females, progeny sex ratio was independent of environmental conditions. Environmental conditions and age should be considered when studying sex ratio bias, which appears to be a function of maternal state rather than of maternal condition. We suggest that a conservative reproductive strategy drives progeny sex ratio in older females according to the "cost of reproduction hypothesis." By manipulating offspring sex ratio, older females reduced the cost of reproduction and increased their expected fitness returns. PMID- 21618924 TI - A test of the mismatch hypothesis: How is timing of reproduction related to food abundance in an aerial insectivore? AB - In seasonal environments, vertebrates are generally thought to time their reproduction so offspring are raised during the peak of food abundance. The mismatch hypothesis predicts that reproductive success is maximized when animals synchronize their reproduction with the food supply. Understanding the mechanisms influencing the timing of reproduction has taken on new urgency as climate change is altering environmental conditions during reproduction, and there is concern that species will not be able to synchronize their reproduction with changing food supplies. Using data from five sites over 24 years (37 site-years), we tested the assumptions of the mismatch hypothesis in the Tree Swallow (Tachycineta bicolor), a widespread aerial insectivore, whose timing of egg laying has shifted earlier by nine days since the 1950s. Contrary to the mismatch hypothesis, the start of egg-laying was strongly related to food abundance (flying insect biomass) during the laying period and not to timing of the seasonal peak in food supply. In fact, food abundance generally continued to increase throughout the breeding season, and there was no evidence of selection based on the mistiming of laying with the seasonal peak of food abundance. In contrast, there was selection for laying earlier, because birds that lay earlier generally have larger clutches and fledge more young. Overall, initial reproductive decisions in this insectivore appear to be based on the food supply during egg formation and not the nestling period. Thus, the mismatch hypothesis may not apply in environments with relatively constant or abundant food throughout the breeding season. Although climate change is often associated with earlier reproduction, our results caution that it is not necessarily driven by selection for synchronized reproduction. PMID- 21618925 TI - A robust-design formulation of the incidence function model of metapopulation dynamics applied to two species of rails. AB - The incidence function model (IFM) uses area and connectivity to predict metapopulation dynamics. However, false absences and missing data can lead to underestimates of the number of sites contributing to connectivity, resulting in overestimates of dispersal ability and turnovers (extinctions plus colonizations). We extend estimation methods for the IFM by using a hierarchical Bayesian model to account both for false absences due to imperfect detection and for missing data due to sites not surveyed in some years. We compare parameter estimates, measures of metapopulation dynamics, and forecasts using stochastic patch occupancy models (SPOMs) among three IFM models: (1) a Bayesian formulation assuming no false absences and omitting site-year combinations with missing data; (2) a hierarchical Bayesian formulation assuming no false absences but incorporating missing data; and (3) a hierarchical Bayesian formulation allowing for imperfect detection and incorporating missing data. We fit the models to multiyear data sets of occupancy for two bird species that differ in body size and presumed dispersal ability but inhabit the same network of sites: the small Black Rail (Laterallus jamaicensis) and the medium-sized Virginia Rail (Rallus limicola). Incorporating missing data affected colonization parameters and led to lower estimates of dispersal ability for the Black Rail. Detection rates were high for the Black Rail in most years but moderate for the Virginia Rail. Incorporating imperfect detection resulted in higher occupancy and lower turnover rates for both species, with largest effects for the Virginia Rail. Forecasts using SPOMs were sensitive to both missing data and false absences; persistence in models assuming no false absences was more optimistic than from robust models. Our results suggest that incorporating false absences and missing data into the IFM can improve (1) estimates of dispersal ability and the effect of connectivity on colonization, (2) the scaling of extinction risk with patch area, and (3) forecasts of occupancy and turnover rates. PMID- 21618926 TI - Metabolic rate throughout the annual cycle reveals the demands of an Arctic existence in great cormorants. AB - Aquatic endotherms living in polar regions are faced with a multitude of challenges, including low air and water temperatures and low illumination, especially in winter. Like other endotherms from cold environments, Great Cormorants (Phalacrocorax carbo) living in Arctic waters were hypothesized to respond to these challenges through a combination of high daily rate of energy expenditure (DEE) and high food requirements, which are met by a high rate of catch per unit effort (CPUE). CPUE has previously been shown in Great Cormorants to be the highest of any diving bird. In the present study, we tested this hypothesis by making the first measurements of DEE and foraging activity of Arctic-dwelling Great Cormorants throughout the annual cycle. We demonstrate that, in fact, Great Cormorants have surprisingly low rates of DEE. This low DEE is attributed primarily to very low levels of foraging activity, particularly during winter, when the cormorants spent only 2% of their day submerged. Such a low level of foraging activity can only be sustained through consistently high foraging performance. We demonstrate that Great Cormorants have one of the highest recorded CPUEs for a diving predator; 18.6 g per minute submerged (95% prediction interval 13.0-24.2 g/min) during winter. Temporal variation in CPUE was investigated, and highest CPUE was associated with long days and shallow diving depths. The effect of day length is attributed to seasonal variation in prey abundance. Shallow diving leads to high CPUE because less time is spent swimming between the surface and the benthic zone where foraging occurs. Our study demonstrates the importance of obtaining accurate measurements of physiology and behavior from free-living animals when attempting to understand their ecology. PMID- 21618927 TI - Top-down control inhibits spatial self-organization of a patterned landscape. AB - Regular, self-organized spatial patterns in primary producers have been described in a wide range of ecosystems and are predicted to affect community production and resilience. Although consumers are abundant in most systems, the effect of trophic interactions on pattern formation in primary producers remains unstudied. We studied the effects of top-down control by herbivores on a self-organized landscape of regularly spaced, diatom-covered hummocks alternating with water filled hollows on an intertidal mudflat in The Netherlands. Spatial patterns developed during spring but were followed by a rapid collapse in summer, leading to a flat landscape with low diatom densities and little variation in sediment bed level. This dramatic decline co-occurred with a gradual increase of benthic herbivores. A manipulative field experiment, where benthic herbivores were removed from the sediment, revealed that both diatom growth and hummock formation were inhibited by the activity of benthic herbivores. Our study provides clear evidence of top-down control of spatial self-organized patterns by benthic herbivores within a biological-geomorphic landscape. PMID- 21618928 TI - Spatial variability of biotic and abiotic tree establishment constraints across a treeline ecotone in the Alaska range. AB - Throughout interior Alaska (U.S.A.), a gradual warming trend in mean monthly temperatures occurred over the last few decades (approximatlely 2-4 degrees C). The accompanying increases in woody vegetation at many alpine treeline (hereafter treeline) locations provided an opportunity to examine how biotic and abiotic local site conditions interact to control tree establishment patterns during warming. We devised a landscape ecological approach to investigate these relationships at an undisturbed treeline in the Alaska Range. We identified treeline changes between 1953 (aerial photography) and 2005 (satellite imagery) in a geographic information system (GIS) and linked them with corresponding local site conditions derived from digital terrain data, ancillary climate data, and distance to 1953 trees. Logistic regressions enabled us to rank the importance of local site conditions in controlling tree establishment. We discovered a spatial transition in the importance of tree establishment controls. The biotic variable (proximity to 1953 trees) was the most important tree establishment predictor below the upper tree limit, providing evidence of response lags with the abiotic setting and suggesting that tree establishment is rarely in equilibrium with the physical environment or responding directly to warming. Elevation and winter sun exposure were important predictors of tree establishment at the upper tree limit, but proximity to trees persisted as an important tertiary predictor, indicating that tree establishment may achieve equilibrium with the physical environment. However, even here, influences from the biotic variable may obscure unequivocal correlations with the abiotic setting (including temperature). Future treeline expansion will likely be patchy and challenging to predict without considering the spatial variability of influences from biotic and abiotic local site conditions. PMID- 21618929 TI - A strong test of a maximum entropy model of trait-based community assembly. AB - We evaluate the predictive power and generality of Shipley's maximum entropy (maxent) model of community assembly in the context of 96 quadrats over a 120-km2 area having a large (79) species pool and strong gradients. Quadrats were sampled in the herbaceous understory of ponderosa pine forests in the Coconino National Forest, Arizona, U.S.A. The maxent model accurately predicted species relative abundances when observed community-weighted mean trait values were used as model constraints. Although only 53% of the variation in observed relative abundances was associated with a combination of 12 environmental variables, the maxent model based only on the environmental variables provided highly significant predictive ability, accounting for 72% of the variation that was possible given these environmental variables. This predictive ability largely surpassed that of nonmetric multidimensional scaling (NMDS) or detrended correspondence analysis (DCA) ordinations. Using cross-validation with 1000 independent runs, the median correlation between observed and predicted relative abundances was 0.560 (the 2.5% and 97.5% quantiles were 0.045 and 0.825). The qualitative predictions of the model were also noteworthy: dominant species were correctly identified in 53% of the quadrats, 83% of rare species were correctly predicted to have a relative abundance of < 0.05, and the median predicted relative abundance of species actually absent from a quadrat was 5 x 10(-5). PMID- 21618930 TI - "The upper limits of vegetation on Mauna Loa, Hawaii": a 50th-anniversary reassessment. AB - In January 1958, a survey of alpine flora was conducted along a recently constructed access road across the upper volcanic slopes of Mauna Loa, Hawaii (2525-3397 m). Only five native Hawaiian species were encountered on sparsely vegetated historic and prehistoric lava flows adjacent to the roadway. A resurvey of roadside flora in 2008 yielded a more than fourfold increase to 22 species, including nine native species not previously recorded. Eight new alien species have now invaded this alpine environment, although exclusively limited to a few individuals in ruderal habitat along the roadway. Alternative explanations for species invasion and altitudinal change over the past 50 years are evaluated: (1) changes related to continuing primary succession on ameliorating (weathering) young lava substrates; (2) local climate change; and (3) road improvements and increased vehicular access which promote enhanced car-borne dispersal of alien species derived from the expanding pool of potential colonizers naturalized on the island in recent decades. Unlike alpine environments in temperate latitudes, the energy component (warming) in climate change on Mauna Loa does not appear to be the unequivocal driver of plant invasion and range extension. Warming may be offset by other climate change factors including rainfall and evapotranspiration. PMID- 21618931 TI - A hierarchical model for spatial capture-recapture data: comment. PMID- 21618932 TI - Magnitude of cement-device interfacial stresses with and without tibial stemming: impact of BMI. AB - Patients expect their total knee arthroplasty to relieve pain and to be long lasting. With patients becoming more active, weighing more, and living longer, this expectation becomes increasingly more difficult to fulfill. Patients who are obese and active put greater loads on their implants and may have a greater risk of failure. Although much attention has been paid to decreasing polyethylene wear, a major cause of implant failure, very little research focus has been directed to elucidate other measures to reduce failure, such as the efficacy of prophylactic stemming of the tibial tray. This study explored whether additional mechanical support for tibial base plates would help reduce bone cement stresses in heavy patients, who, like patients with a high activity level, put added stress on their implants. A tibial base plate with a 12-mm-diameter x 50-mm-long stem was compared with the same tibial base plate with a 15-mm-diameter x 20-mm long end cap using finite element analysis. The results indicate that the tibial base plate with a prophylactic stem significantly reduced compressive and shear stresses on the cement-device interface and therefore may help to reduce the possibility of tibial loosening in these at-risk patients. Further, such studies will aid the surgeon in educating patients and in selecting the appropriate implant strategy. PMID- 21618933 TI - The influence of material and design on total knee replacement wear. AB - It is difficult for surgeons to make the decision on which design or material to use given the different options available. Marketing claims and direct-to consumer advertising certainly complicate this further. One company may claim a higher percentage of wear reduction with their bearing surfaces compared with those of another manufacturer. If the percentage of wear reduction is lower, it is unclear as to whether this creates a more effective technology for reducing wear in the clinical situation. The relative contribution and relationship of design and materials to wear performance must be considered before making that important judgment. To examine the overall influence of implant design on wear reduction, a knee simulator study was undertaken. This simulator study compared the Oxinium Genesis II system with the Triathlon Conventional and Triathlon X3 knee systems under physiologic stair-climb loading and motion profiles. This allows a similar comparison of material effect within one design but also a global comparison across designs. Test results show the Triathlon Conventional and Triathlon X3 knee systems have superior wear resistance compared with that of the Genesis II Oxinium system under stair-climbing simulation. This finding indicates that implant design plays a more significant role in knee wear reduction than material. Although material technology may improve a given knee system's ability to wear, design geometry has a first-order effect and should be addressed before materials. This study represents an effort to differentiate design effect from two different approaches to material enhancement. The results of this study support the predominance of design in knee replacement wear performance. Ultimately, only clinical evidence such as published studies or outcomes reported in the available joint registries will establish whether any material or design can achieve a 30-year outcome. PMID- 21618934 TI - Maximizing patient satisfaction and functional results after total knee arthroplasty. AB - The purpose of this article is to review the issues regarding preoperative patient expectations for total knee arthroplasty and whether or not these are fulfilled after surgery. The demographics of the typical total knee arthroplasty patient are changing, and the expectations regarding the outcome of the surgery have been changing as well. Patients are younger, heavier, more active, and often come to the surgeon's office with information provided via the Internet that may be inaccurate and/or misleading. Many expect and assume that the operation will return the knee to "normal." Traditional outcomes measures used by orthopedic surgeons to determine the quality of the results achieved are inadequate and do not take into account the higher expectations of current patients. Various strategies have been developed to improve satisfaction after the surgery. These include preoperative patient education, less invasive surgical approaches, advances in prosthetic design, multimodal pain management, and aggressive postoperative rehabilitation. Using these strategies will make preoperative expectations more realistic and improve postoperative satisfaction. PMID- 21618935 TI - Who needs autologous blood donation in joint replacement? AB - Conflicting results are available about the efficacy of routine preoperative autologous donation (PAD) in reducing allogenic blood transfusion during total joint arthroplasty (TJA). This study aimed to determine if PAD is effective in reducing the need for allogenic transfusion after TJA. For this retrospective study, data on 409 patients who received total knee arthroplasty (TKA) and 513 who underwent total hip arthroplasty (THA) from January to June 2005 were evaluated. It is our institutional policy to offer preoperative donation to all patients. Based on patient decision, preoperative hemoglobin, and other factors, PAD may or may not take place. Five hundred forty-six (61%) patients donated on average 1.3 units (range, 1 to 2) of blood. Autologous and allogenic transfusions were respectively performed in 91% (514 patients) and 24% of our cohort. The rate of allogenic transfusion after TKA was lower in the PAD group at 21% versus 27% among the nondonors, although it was not statistically significant (p = 0.10). The allogenic transfusion rate after THA was significantly lower among autologous donors (16% versus 34%, p = 0.003). Advanced age, lower body mass index, simultaneous bilateral arthroplasty, and lower preoperative hemoglobin were independently associated with increased allogenic blood transfusion. PAD seems to be effective in reducing allogenic transfusions after THA but not TKA. PMID- 21618936 TI - Angioleiomyoma of the knee: case series and an unusual cause of knee pain. AB - We present a consecutive series of five patients with angioleiomyoma, a rare benign soft tissue tumor around the knee. Between 2005 and 2010, five patients with histopathologically confirmed diagnosis of angioleiomyoma of the knee underwent surgical excision. All five patients were Afro-caribbean in origin. The presenting feature was burning medial knee pain localized to a tender hyperesthetic swelling around the knee. The mean age of presentation was 45 years (range, 42 to 51 years), and the mean duration of symptom was 34 months (range, 12 to 96 months). Although magnetic resonance imaging (MRI) scans were helpful in locating and delineating the tumors, true diagnosis was established only by histopathology. All patients underwent surgical excision with complete resolution of symptoms. No recurrences were seen at an average follow-up of 16 months (range, 3 to 24 months). The treatment of these tumors, especially around the knee, is frequently delayed due to their late presentation and lack of awareness of this clinical condition. We recommend a high index of suspicion in patients, especially of Afro-caribbean origin, presenting with painful hyperesthetic subcutaneous swelling around the knee. Early investigation by MRI expedites the diagnosis, and surgical excision results in complete resolution of symptoms. PMID- 21618937 TI - No correlation of height or gender with anterior cruciate ligament footprint size. AB - Recently, there has been much interest in anatomic double-bundle reconstruction of the anterior cruciate ligament (ACL). Double-bundle reconstruction of the ACL requires adequate footprint size to place two femoral tunnels. The purpose of this study was to determine if there is a correlation between lateral intercondylar ridge length and gender and/ or height. We measured the femoral attachment of the ACL to determine if patient sex and/or height could be used to predict ACL femoral footprint size. We measured the length of the lateral intercondylar ridge in 65 skeletally mature human femora. Gender and height was recorded for each individual. We used bivariate regression analysis to determine correlations between both height and gender and the length of the lateral intercondylar ridge. The principal findings of our study demonstrate that there is no correlation between ACL femoral footprint size and gender or footprint size and height. Our study demonstrates that patient height and gender cannot be used for preoperative planning when deciding whether a given patient has adequate footprint size to support double-bundle reconstruction of the ACL. PMID- 21618938 TI - Biomechanical evaluation of a high tibial osteotomy with a meniscal transplant. AB - This study determines the biomechanical advantage and the optimal configuration of a high tibial osteotomy (HTO) and meniscus transplantation performed concurrently. Six cadaver knees were placed in a spatial frame, and an HTO was completed. Loading points between a mechanical 6 degrees of varus and 8 degrees ofvalgus were loaded to 800 N for medial meniscal intact, meniscectomized, and transplanted states. Posterior slope was also increased by 3 degrees in these specimens. Contact data was recorded. Peak pressures significantly increased in the meniscectomized state in every degree of varus/valgus (p < 0.05). For both peak and total medial compartment pressures, there was a significant drop (p < 0.001) between neutral and 3 degrees of valgus. Lateral compartment pressures linearly increased from varus to valgus orientation. There was no significant change in the pressure profile of the knee with a 3-degree increase in posterior slope. This biomechanical study confirms the hypothesis that an HTO improves the peak pressures in the medial compartment at all degrees of varus/valgus alignment in the setting of meniscal transplantation. Furthermore, the largest decrease in medial pressures was between neutral and 3 degrees of valgus, suggesting that perhaps neutral aligned knees could benefit from an HTO. PMID- 21618939 TI - Water-tight knee arthrotomy closure: comparison of a novel single bidirectional barbed self-retaining running suture versus conventional interrupted sutures. AB - Standard medial parapatellar arthrotomies of 10 cadaveric knees were closed with either conventional interrupted absorbable sutures (control group, mean of 19.4 sutures) or a single running knotless bidirectional barbed absorbable suture (experimental group). Water-tightness of the arthrotomy closure was compared by simulating a tense hemarthrosis and measuring arthrotomy leakage over 3 minutes. Mean total leakage was 356 mL and 89 mL in the control and experimental groups, respectively (p = 0.027). Using 8 of the 10 knees (4 closed with control sutures, 4 closed with an experimental suture), a tense hemarthrosis was again created, and iatrogenic suture rupture was performed: a proximal suture was cut at 1 minute; a distal suture was cut at 2 minutes. The impact of suture rupture was compared by measuring total arthrotomy leakage over 3 minutes. Mean total leakage was 601 mL and 174 mL in the control and experimental groups, respectively (p = 0.3). In summary, using a cadaveric model, arthrotomies closed with a single bidirectional barbed running suture were statistically significantly more water tight than those closed using a standard interrupted technique. The sample size was insufficient to determine whether the two closure techniques differed in leakage volume after suture rupture. PMID- 21618940 TI - Clinical benefits of intra-articular anakinra for persistent knee effusion. AB - Postoperative or postinjury inflammation after knee surgery is a significant clinical challenge. Cytokines, particularly interleukin-1 (IL-1), released by inflammatory cells have been shown to have a role in cartilage homeostasis and tissue repair and fibrosis. Anakinra, an IL-1 receptor antagonist (IL-1ra), has shown promising results from its use in treating rheumatoid arthritis, juvenile inflammatory arthritis, and autoimmune inflammatory syndromes. We hypothesized that intra-articular anakinra injection may result in sustained improvements of persistent effusion of the knee joint refractory to other modalities. We retrospectively reviewed 6 patients (3 female, 3 male), ranging in age from 17 to 50 years, who underwent injection of intra-articular anakinra, 200 mg, between November 15, 2007, and April 29, 2010, for persistent effusions of the postoperative knee. All of the patients treated with intra-articular anakinra for persistent effusions failed conservative treatment with physical therapy, oral anti-inflammatory medication, and 4 of 6 patients failed prior corticosteroid injections. After intra-articular anakinra, 66% had improvement in knee arc of motion (15 to 30 degrees) and pain, and 5 of 6 (83%) had improvement in swelling. All of these patients were able to return to sports. We found intra-articular anakinra to be safe and effective in this small group of patients with persistent effusions. There were no adverse clinical reactions or infections. These findings provide support for further study of IL-1 inhibition in the management of postsurgical inflammation. PMID- 21618941 TI - Bilateral complete discoid medial meniscus: a case report. AB - Discoid medial meniscus is very rare, and bilateral discoid medial meniscus is extremely rare. We report a case of bilateral discoid medial meniscus. The patient had symptoms in the right knee but was asymptomatic in the left knee. We performed magnetic resonance imaging for both knees. The patient was treated by saucerization and by all inside repair of the anterior horn tear in the right knee. We performed second-look arthroscopy after 18 months. PMID- 21618942 TI - Exposures to high levels of carbon monoxide from wood-fired temazcal (steam bath) use in highland Guatemala. AB - The temazcal is a wood-fired steam bath used in the rural highlands of Guatemala for bathing and healing. We measured carbon monoxide (CO) among 288 participants in 72 temazcales. Participants were drawn from communities who participated in the RESPIRE (Randomized Exposure Study of Pollution Indoors and Respiratory Effects) chimney stove intervention trial. Temazcal CO exposures were extremely high, averaging 431 parts per million (time-weighted average). Compared to kitchen wood-smoke exposures, the temazcal contributes significantly to weekly exposures, despite the fact that the population spends less time in the temazcal than in the kitchen. This report 1) describes temazcal use patterns; 2) reports participants' signs and symptoms during temazcal use; 3) models the distribution of temazcal CO concentrations; 4) assesses reliability of exhaled breath CO as a biomarker of CO exposure; and 5) provides a proportional analysis of CO concentrations from temazcal use, as compared to kitchen concentrations. PMID- 21618943 TI - A baseline evaluation of traditional cook stove smoke exposures and indicators of cardiovascular and respiratory health among Nicaraguan women. AB - Biomass-derived indoor air pollution has been associated with increased risks of respiratory diseases; however, relatively few studies have examined the cardiovascular effects of biomass burning. We measured 48-hour indoor fine particulate matter and indoor and personal carbon monoxide (CO) concentrations in 124 households using open-fire cook stoves in Nicaragua. We also examined the cross-sectional relationship of air pollution and health. High air pollutant concentrations with considerable variability were measured. Nonsignificant elevations in systolic blood pressure were associated with increases in CO concentrations. These associations were stronger among obese participants; an 8.51 mmHg (95% confidence interval [CI]: 3.06, 13.96) increase in systolic blood pressure per 24 ppm increase in 48-hour average indoor CO levels was observed. Although the cross-sectional design of this study limits the interpretation, we observed evidence of a relationship between indoor air pollution and blood pressure and heart rate, two indicators of cardiovascular health. PMID- 21618944 TI - A proposal for a safe exposure level for diacetyl. AB - Diacetyl is a naturally occurring compound that has been used in concentrated form as a food additive, particularly in butter flavorings. Inhalation of diacetyl and butter flavoring fumes has caused a variety of respiratory diseases in workers and consumers including bronchiolitis obliterans (BO), a relatively rare, severe, and irreversible lung disease. A safe level of exposure to diacetyl has not been established. We review the literature on diacetyl and flavoring toxicity and critique a recent proposal for an occupational exposure limit (OEL) of 0.2 ppm for diacetyl. We present unpublished data and novel analyses in support of our proposal for a safe level of exposure. Our findings indicate that a safe level of exposure exists around or below a time-weighted average of 1 ppb for an eight-hour workday. The levels of exposure we found to be unsafe include ranges that popcorn consumers may potentially be exposed to, indicating a risk of severe lung disease (including BO) for some consumers. PMID- 21618945 TI - Neurological evaluation of newborn infants of mothers working in citrus groves in Northern Thailand. AB - The purpose of this pilot study was to assess neurological function in newborn infants born to mothers working in citrus orchards in Northern Thailand for a period in excess of one year where pesticide applications average 35 times a year. Forty-one infants from uncomplicated term births at the community hospital in Fang, Thailand, were given neurological evaluations during the first four days of life. This was a cross-sectional study in which nine mothers worked in citrus orchards and 32 mothers did not. Examiners were not given the exposure history of the mothers. Twelve infants--five of them born to mothers from citrus orchards- had examinations demonstrating abnormal muscle tone. Using logistic regression, the final model showed that maternal citrus grove exposure and anesthesia use were significant predictors (p < 0.05) of abnormal muscle tone with adjusted odds ratios of 9.82 (CI = 1.42, 68.07) and 5.99 (CI = 1.003, 35.85) for exposure and anesthesia respectively. PMID- 21618946 TI - Asbestos-related disease in Bangladeshi ship breakers: a pilot study. AB - A pilot study tested the feasibility of conducting occupational health research in Bangladesh while examining prevalence of asbestos-related diseases including asbestosis, work-related respiratory symptoms, and attitudes to occupational health and safety among a group of internal migrant ship breakers. Data was collected on clinical and work history, respiratory symptoms, and occupational health and safety practices in Bengali. A B-reader read all postero-anterior chest x-rays. In the 104 male ship breakers studied, prevalence of asbestos related disease was 12%, of which asbestosis accounted for 6%. Knowledge of asbestos and occupational health and safety measures were almost nonexistent. The prevalence of asbestos-related diseases is low compared to studies in shipbuilders and repairers, but a risk underestimate could have resulted from challenges identified during study design and implementation including: industry noncooperation and a culture of corruption; technological and language barriers; and a regional lack of physician knowledge and research on occupational diseases. PMID- 21618947 TI - Shift work and the risk of metabolic syndrome: a nested case-control study. AB - The aim of the present study was to examine the association between shift work and the metabolic syndrome (MetS) using a large-scale longitudinal study design. Data were collected from a historical cohort of health checkups in the Japanese population. The baseline survey, which involved 16,952 inhabitants of the Minami Saku area of the Nagano Prefecture, was started in 1978. A nested case-control study was conducted between 1987 and 1990. This analysis was restricted to 6,712 men and women (age range 25-59 years). A conditional logistic regression model was used to estimate the risk of MetS associated with shift work. Compared with the day workers, shift workers had a significantly higher risk of MetS (odds ratio 1.87; 95% CI, 1.13-3.08). Our results demonstrate that shift work was strongly associated with MetS. The study suggests appropriate dietary habits as a basis for managing the MetS risk of shift workers. PMID- 21618948 TI - The adverse health effects of oil spills: a review of the literature and a framework for medically evaluating exposed individuals. AB - In April 2010, an explosion on an oil rig in the Gulf of Mexico killed 11 workers, injured 17 workers, and spilled an estimated 185 million gallons of crude oil into the Gulf. Adverse effects on the health of cleanup workers, fishermen, and others as well as on the ecosystem are being studied. This paper reviews published studies of the adverse health effects due to previous oil spills. Acute effects have included: respiratory, eye, and skin symptoms; headache; nausea; dizziness; and tiredness or fatigue. Chronic effects have included: psychological disorders, respiratory disorders, genotoxic effects, and endocrine abnormalities. We also present a systematic approach to evaluating individuals exposed to oil spills. PMID- 21618949 TI - Ghostwriting in medical literature. PMID- 21618951 TI - Corporate versus public interests: community responsibility to defend scientific integrity. PMID- 21618952 TI - Open letter to Normand Paulin, Occupational Health & Safety Commission of Quebec, on the export of asbestos from Canada. PMID- 21618953 TI - The real CER: lost in translation. PMID- 21618954 TI - Management strategies in acute lymphoblastic leukemia. AB - Survival in acute lymphoblastic leukemia (ALL) has improved in recent decades due to recognition of the biologic heterogeneity of ALL, utilization of risk-adapted therapy, and development of protocols that include optimized chemotherapy combinations, effective central nervous system (CNS) prophylaxis, post-induction intensification of therapy, and a prolonged maintenance phase of treatment. Recent molecular studies have yielded novel insights into both leukemia biology and host pharmacogenetic factors; also, large cooperative group clinical research studies have successively refined effective treatment strategies. While children have higher remission and cure rates than adults, both populations have benefited from these discoveries and innovations. Future challenges in this field include improving outcomes for high-risk patients and those with relapsed disease, and developing and integrating novel targeted therapeutic agents into current regimens to reduce toxicities while further improving outcomes. PMID- 21618955 TI - What pediatrics can teach us. PMID- 21618956 TI - Recent advances in acute lymphoblastic leukemia. PMID- 21618957 TI - Diagnosis of invasive lobular carcinoma in a young woman presenting with pleomorphic lobular carcinoma in situ on core biopsy. PMID- 21618958 TI - Pleomorphic lobular carcinoma in situ: A divergent entity with emerging significance. PMID- 21618959 TI - The impact of lobular histology on breast cancer treatment. AB - Lobular neoplasias are a distinct clinical entity with subtle differences in locoregional treatment considerations when compared with ductal cancers. Although overall surgical recommendations do not differ significantly between breast cancers of lobular versus ductal histology, there are important distinctions that should be considered as part of patient care, particularly with respect to recommendations regarding management of the contralateral breast and genetic testing. Because the lobular subtype of breast cancer is underrepresented in studies of molecular prognostic markers, the results of such testing must be interpreted with caution until they are validated specifically in patients with lobular histology. Until then, the mainstay of sound treatment decision-making remains a thorough clinical understanding of the disease and of the factors that can have an impact on outcome. PMID- 21618960 TI - How we treat tumor lysis syndrome. AB - Tumor lysis syndrome (TLS) is an oncology emergency that occurs as a result of rapid tumor cell breakdown and the consequent release of massive amounts of intracellular contents, including potassium, phosphate, and uric acid, into the systemic circulation. These metabolic disturbances lead to life-threatening conditions and may cause sudden death if not treated. TLS commonly occurs following initiation of cytotoxic treatment in patients with high-grade lymphomas or acute lymphoblastic leukemia. Spontaneous cases involving both solid and hematologic tumors have also been reported. Rarely, TLS occurs following treatment with irradiation, corticosteroids, hormonal therapy, or biologic therapy. It is necessary to identify patients at risk for TLS early in order to initiate preventive measures. In the event that preventive measures fail, the clinical parameters and signs of TLS must be understood and recognized so that treatment can begin as soon as possible, as this condition is a significant cause of morbidity and mortality. PMID- 21618961 TI - The evolving world of tumor lysis syndrome. PMID- 21618962 TI - The more things change, the more they stay the same. PMID- 21618963 TI - Kava (Piper methysticum). PMID- 21618964 TI - Palladium-catalyzed carbon-monoxide-free aminocarbonylation of aryl halides using N-substituted formamides as an amide source. AB - A carbon-monoxide-free aminocarbonylation of various N-substituted formamides with aryl iodides and aryl bromides using palladium acetate and Xantphos is described. The developed methodology is applicable for a wide range of formamides and aryl halides containing different functional groups furnishing good to excellent yield of the corresponding products. N-substituted formamides are used as an amide source wherein a Vilsmeier-type intermediate plays a major role, thus eliminating the need of toxic carbon monoxide gas. PMID- 21618965 TI - Antennal proteome comparison of sexually mature drone and forager honeybees. AB - Honeybees have evolved an intricate system of chemical communication to regulate their complex social interactions. Specific proteins involved in odorant detection most likely supported this chemical communication. Odorant reception takes place mainly in the antennae within hairlike structures called olfactory sensilla. Antennal proteomes of sexually mature drone and forager worker bees (an age group of bees assigned to perform field tasks) were compared using two dimensional electrophoresis, mass spectrometry, quantitative real-time polymerase chain reaction, and bioinformatics. Sixty-one differentially expressed proteins were identified in which 67% were highly upregulated in the drones' antennae whereas only 33% upregulated in the worker bees' antennae. The antennae of the worker bees strongly expressed carbohydrate and energy metabolism and molecular transporters signifying a strong demand for metabolic energy and odorant binding proteins for their foraging activities and other olfactory responses, while proteins related to fatty acid metabolism, antioxidation, and protein folding were strongly upregulated in the drones' antennae as an indication of the importance for the detection and degradation of sex pheromones during queen identification for mating. On the basis of both groups of altered antenna proteins, carbohydrate metabolism and energy production and molecular transporters comprised more than 80% of the functional enrichment analysis and 45% of the constructed biological interaction networks (BIN), respectively. This suggests these two protein families play crucial roles in the antennal olfactory function of sexually mature drone and forager worker bees. Several key node proteins in the BIN were validated at the transcript level. This first global proteomic comparative analysis of antennae reveals sex-biased protein expression in both bees, indicating that odorant response mechanisms are sex-specific because of natural selection for different olfactory functions. To the best of our knowledge, this result further provides extensive insight into the expression of the proteins in the antennae of drone and worker honeybees and adds vital information to the previous findings. It also provides a new angle for future detailed functional analysis of the antennae of the honeybee castes. PMID- 21618966 TI - Isolation, chemical, and biotransformation routes of labdane-type diterpenes. PMID- 21618967 TI - Uranium sorption on various forms of titanium dioxide--influence of surface area, surface charge, and impurities. AB - Titanium dioxide (TiO(2)) has often served as a model substrate for experimental sorption studies of environmental contaminants. However, various forms of Ti oxide have been used, and the different sorption properties of these materials have not been thoroughly studied. We investigated uranium sorption on some thoroughly characterized TiO(2) surfaces with particular attention to the influence of surface area, surface charge, and impurities. The sorption of U(VI) differed significantly between samples. Aggressive pretreatment of one material to remove impurities significantly altered the isoelectric point, determined by an electroacoustic method, but did not significantly impact U sorption. Differences in sorption properties between the various TiO(2) materials were related to the crystallographic form, morphology, surface area, and grain size, rather than to surface impurities or surface charge. In-situ attenuated total reflection Fourier-transform infrared (ATR FT-IR) spectroscopic studies showed that the spectra of the surface species of the TiO(2) samples are not significantly different, suggesting the formation of similar surface complexes. The data provide insights into the effect of different source materials and surface properties on radionuclide sorption. PMID- 21618968 TI - Molecular weight change of polyhydroxyalkanoate (PHA) caused by the PhaC subunit of PHA synthase from Bacillus cereus YB-4 in recombinant Escherichia coli. AB - Polyhydroxyalkanoate (PHA)-producing Bacillus strains possess class IV PHA synthases composed of two subunit types, namely, PhaR and PhaC. In the present study, PHA synthases from Bacillus megaterium NBRC15308(T) (PhaRC(Bm)), B. cereus YB-4 (PhaRC(YB4)), and hybrids (PhaR(Bm)C(YB4) and PhaR(YB4)C(Bm)) were expressed in Escherichia coli JM109 to characterize the molecular weight of the synthesized poly(3-hydroxybutyrate) [P(3HB)]. PhaRC(Bm) synthesized P(3HB) with a relatively high molecular weight (M(n) = 890 * 10(3)) during 72 h of cultivation, whereas PhaRC(YB4) synthesized low-molecular-weight P(3HB) (M(n) = 20 * 10(3)). The molecular weight of P(3HB) synthesized by PhaRC(YB4) decreased with increasing culture time and temperature. This time-dependent behavior was observed for hybrid synthase PhaR(Bm)C(YB4), but not for PhaR(YB4)C(Bm). These results suggest that the molecular weight change is caused by the PhaC(YB4) subunit. The homology between PhaCs from B. megaterium and B. cereus YB-4 is 71% (amino acid identity); however, PhaC(YB4) was found to have a previously unknown effect on the molecular weight of the P(3HB) synthesized in E. coli. PMID- 21618969 TI - Total synthesis of bryostatin 9. AB - The total synthesis of bryostatin 9 was accomplished using a uniquely step economical and convergent Prins-driven macrocyclization strategy. At 25 linear and 42 total steps, this is currently the most concise and convergent synthesis of a potent bryostatin. PMID- 21618970 TI - Fluorescent proton sensors based on energy transfer. AB - Photophysical data and orbital energy levels (from electrochemistry) were compared for molecules with the same BODIPY acceptor part (red) and perpendicularly oriented xanthene or BODIPY donor fragments (green). Transfer of energy, hence the photophysical properties of the cassettes, including the pH dependent fluorescence in the xanthene-containing molecules, correlates with the relative energies of the frontier orbitals in these systems. Intracellular sensing of protons is often achieved via sensors that switch off completely at certain pH values, but probes of this type are not easy to locate inside cells in their "off-state". A communication from these laboratories (J. Am. Chem. Soc., 2009, 131, 1642-3) described how the energy transfer cassette 1 could be used for intracellular imaging of pH. This probe is fluorescent whatever the pH, but its exact photophysical properties are governed by the protonation states of the xanthene donors. This work was undertaken to further investigate correlations between structure, photophysical properties, and pH for energy transfer cassettes. To achieve this, three other cassettes 2-4 were prepared: another one containing pH-sensitive xanthene donors (2) and two "control cassettes" that each have two BODIPY-based donors (3 and 4). Both the cassettes 1 and 2 with xanthene based donors fluoresce red under slightly acidic conditions (pH < ~6) and green when the medium is more basic (>~7), whereas the corresponding cassettes with BODIPY donors give almost complete energy transfer regardless of pH. The cassettes that have BODIPY donors, by contrast, show no significant fluorescence from the donor parts, but the overall quantum yields of the cassettes when excited at the donor (observation of acceptor fluorescence) are high (ca. 0.6 and 0.9). Electrochemical measurements were performed to elucidate orbital energy level differences between the pH-fluorescence profiles of cassettes with xanthene donors, relative to the two with BODIPY donors. These studies confirm energy transfer in the cassettes is dramatically altered by analytes that perturb relative orbital levels. Energy transfer cassettes with distinct fluorescent donor and acceptor units provide a new, and potentially useful, approach to sensors for biomedical applications. PMID- 21618971 TI - Discovery of novel Pim-1 kinase inhibitors by a hierarchical multistage virtual screening approach based on SVM model, pharmacophore, and molecular docking. AB - In this investigation, we describe the discovery of novel potent Pim-1 inhibitors by employing a proposed hierarchical multistage virtual screening (VS) approach, which is based on support vector machine-based (SVM-based VS or SB-VS), pharmacophore-based VS (PB-VS), and docking-based VS (DB-VS) methods. In this approach, the three VS methods are applied in an increasing order of complexity so that the first filter (SB-VS) is fast and simple, while successive ones (PB-VS and DB-VS) are more time-consuming but are applied only to a small subset of the entire database. Evaluation of this approach indicates that it can be used to screen a large chemical library rapidly with a high hit rate and a high enrichment factor. This approach was then applied to screen several large chemical libraries, including PubChem, Specs, and Enamine as well as an in-house database. From the final hits, 47 compounds were selected for further in vitro Pim-1 inhibitory assay, and 15 compounds show nanomolar level or low micromolar inhibition potency against Pim-1. In particular, four of them were found to have new scaffolds which have potential for the chemical development of Pim-1 inhibitors. PMID- 21618972 TI - Reversible on/off conductance switching of single diarylethene immobilized on a silicon surface. AB - The chemical functionalization of hydrogen-terminated silicon(111) surfaces with photochromic diarylethene using an ethylene anchoring group was achieved. Conductive atomic force microscopy measurements showed the current changes on modified silicon electrodes caused by light-induced isomerization of the diarylethene between an open form and a closed form. PMID- 21618973 TI - Analysis of the singlet-triplet splitting computed by the density functional theory-broken-symmetry method: is it an exchange coupling constant? AB - We derive an analytical expression of the density functional theory (DFT)-broken symmetry (BS) estimation J(BS) of the singlet-triplet gap at the "3 sites-4 electrons" level, that is, two S = (1)/(2) metallic sites + one diamagnetic bridge orbital. As originally designed by Noodleman and Davidson (Chem. Phys.1986, 109, 131), J(BS) contains the residual ferromagnetic contribution, single ligand-to-metal and metal-to-metal charge-transfer terms, but no double ligand-to-metal charge-transfer terms or intra/interligand spin-polarization terms. As revealed by the present analysis, the triplet and BS states computed by DFT differ, not only perturbatively (as expected) because of the various physical mechanisms involved (i.e., differential charge-transfer terms) but mainly because of a spurious and unphysical symmetry breaking of the bridge orbitals in the BS state. We examine the consequences of such a difference by deriving two analytical expressions of the exchange coupling constant, one from the BS orbitals designed to match J(BS) and another one from triplet orbitals only. Following and extending on the first paper in the series (J. Phys. Chem. A 2010, 114, 6149), we propose a simple procedure to extract appropriate parameters filling in our analytical expressions. Moreover, we derive the equivalent "3 sites-4 electrons" exchange coupling constant in the configuration-interaction approach, J(CI), for the purpose of comparison. These analytical expressions have been applied to various copper dimers and compared to experimental values. PMID- 21618974 TI - High-temperature behavior and polymorphism in novel members of the perovskite family Pb2LnSbO6 (Ln=Ho, Er, Yb, Lu). AB - The synthesis, crystal structure, and dielectric properties of four novel members of the family of double perovskites Pb(2)LnSbO(6) are described. The room temperature crystal structures were refined from neutron powder diffraction (NPD) data in the monoclinic C2/c (No. 15) space group. They contain a completely ordered array of alternating LnO(6) and SbO(6) octahedra sharing corners, tilted in antiphase along the three pseudocubic axes, with a a(-)b(-)b(-) tilting scheme, which is very unusual in the crystallochemistry of perovskites. The lead atoms occupy highly asymmetric voids with 8-fold coordination due to the stereoactivity of the Pb(2+) electron lone-pair. Several trends are observed for the entire family of compounds upon heating. The Ln = Lu, Yb, and Er oxides display three successive phase transitions in a narrow temperature range, as shown by differential scanning calorimetry (DSC) data, while the Ln = Ho shows only two transitions. Different crystal structure evolutions have been found from temperature-dependent NPD and DSC, following the space-group sequence C2/c -> P2(1)/n -> R 3 -> Fm 3m for Ln = Lu and Yb, the sequence C2/c -> unknown -> P2(1)/n -> Fm 3m for Ln = Er, and C2/c -> P2(1)/n -> Fm 3m for Ln = Ho. The Ln/Sb long-range ordering is preserved across the consecutive phase transitions. Dielectric permittivity measurements indicate the presence of a paraelectric/antiferroelectric transition (associated with the last structural transition), as suggested by the negative Curie temperature from the Curie-Weiss fit of the reciprocal permittivity. PMID- 21618975 TI - Optical and photocatalytic properties of heavily F(-)-doped SnO2 nanocrystals by a novel single-source precursor approach. AB - Heavily F-doped SnO(2) nanocrystals were successfully prepared by a novel synthetic approach involving low-temperature oxidation of a Sn(2+)-containing fluoride complex KSnF(3) as the single-source precursor with H(2)O(2). The F doped SnO(2) powder was characterized by powder X-ray diffraction, TG-MS, BET surface area, diffuse reflectance spectroscopy, XPS, PL, FTIR spectroscopy, Raman spectroscopy, EPR spectroscopy, SEM, and TEM. Broadening of the diffracted peaks, signifying the low crystallite size of the products, was quite evident in the powder X-ray diffraction pattern of SnO(2) obtained from KSnF(3). It was indexed in a tetragonal unit cell with lattice constants a = 4.7106 (1) A and c = 3.1970 (1) A. Agglomeration of particles, with an average diameter of 5-7 nm, was observed in the TEM images whose spotwise EDX analysis indicated the presence of fluoride ions. In the core level high-resolution F 1s spectrum, the peak observed at 685.08 eV was fitted by the Gaussian profile yielding the fluoride ion concentration to be 21.23% in the SnO(2) lattice. Such a high fluoride ion concentration is reported for the first time in powders. SnO(2):F nanocrystals showed greater thermal stability up to 300 degrees C when heated in a thermobalance under flowing helium, after which generation of small quantities of HF was observed in the TG coupled mass spectrometry analysis. The band gap value, estimated from the Kubelka-Munk function, showed a large shift from 3.52 to 3.87 eV on fluoride ion doping, as observed in the diffuse reflectance spectrum. Such a large shift was corroborated to the overdoped situation due to the Moss Burstein effect with an increase in the carrier concentration. In the photoluminescence (PL) spectrum, SnO(2):F nanocrystals exhibited a broad green emission arising from the singly ionized oxygen vacancies created due to higher dopant concentration. The evidence for singly ionized vacancies was arrived from the presence of a signal with a g value of 1.98 in the ESR spectrum of SnO(2):F at room temperature. The disordered nature of the rutile lattice and the enormous oxygen vacancies created due to fluoride ion doping were evident from the broad bands observed at 455, 588, and 874 cm(-1) in the room-temperature Raman spectrum of SnO(2):F. As the consequence of the oxygen vacancies, F-doped SnO(2) was examined for the function as a photocatalyst in the degradation of aqueous RhB dye solution under UV irradiation. A very high photocatalytic efficiency was observed for the F-doped SnO(2) nanocrystals as compared to pure SnO(2). The BET surface area of pure SnO(2) was quite high (207.81 m(2)/g) as compared to the F doped SnO(2) nanocrystals (45.16 m(2)/g). Pore size analysis showed a mean pore diameter of 1.97 and 13.97 nm for the pure and doped samples. The increased photocatalytic efficiency was related to the very high concentration of oxygen vacancies in SnO(2) induced by F doping. PMID- 21618976 TI - Simultaneous determination of the adsorption constant and the photoinduced electron transfer rate for a CdS quantum dot-viologen complex. AB - Transient absorption (TA) spectroscopy of solution-phase mixtures of colloidal CdS quantum dots (QDs) with acid-derivatized viologen molecules, N-[1-heptyl],N' [3-carboxypropyl]-4,4'-bipyridinium dihexafluorophosphate (V(2+)), indicates electron transfer occurs from the conduction band of the QD to the LUMO of V(2+) after photoexcitation of a band-edge exciton in the QD. Analysis of the magnitude of the ground state bleach of the QD as a function of the molar ratio QD:V(2+) yields the QD-ligand adsorption constant, K(a) (4.4 * 10(4) M(-1)) for V(2+) ligands adsorbed in geometries conducive to electron transfer. The value of K(a), together with the measured rates of (i) formation of the V(+*) electron transfer product and (ii) recovery of the ground state bleach of the QD, enables determination of the intrinsic rate constant for charge separation, k(CS,int) ~ 1.7 * 10(10) s(-1), the rate for a single QD-V(2+) donor-acceptor pair. This analysis confirms previous reports that the number of ligands adsorbed to each QD is well-described by a Poisson distribution. This is the first report where the QD-ligand charge transfer and binding equilibria are quantitatively investigated simultaneously with a single technique. PMID- 21618977 TI - Coexistence of intramolecular ligand-mediated and through hydrogen-bond magnetic interactions in a chain of dicopper(II) units. AB - The reaction of 2,8-dimethyl-5,11-bis(pyridin-2-ylmethyl)-1,4,5,6,7,10,11,12 octahydroimidazo[4,5-h]imidazo[4,5-c][1,6]-diazecine (dimp) with copper(II) nitrate in water produces the compound [Cu(2)(dimp)(H(2)O)(2)(NO(3))(2)](NO(3))(2). The single-crystal X-ray structure shows the formation of hydrogen-bonded chains in the lattice that are formed by dicopper(II) units doubly connected by nitrate/water bridges. Within the one dimensional chains, the Cu ions are separated by either intramolecular or intermolecular distances of 7.309(2) A or 6.255(2) A, respectively. The magnetic susceptibility data revealing weak antiferromagnetic exchange interactions between the copper(II) ions were interpreted by considering two possible models, namely, an isolated dinuclear and a 1-D chain picture. The latter leads to an alternation J(1) = -11.6 and J(2) = -3.0 cm(-1) along the chain. In order to clarify the relative strengths of the exchange couplings through hydrogen bonds and via the bridging dimp ligand, solution EPR studies and quantum chemical calculations were carried out. EPR studies unambiguously conclude on the existence of an exchange interaction J(a) mediated by the dinucleating dimp ligand, while the through-H coupling J(b) is physically absent in solution. On the basis of dinuclear units extracted from the X-ray data, J(a) was estimated around -5.0 cm(-1) from DFT-based calculations (M06 functional), whereas J(b) is negligible. In contrast, wave function configuration interaction calculations (DDCI) support a description where both inter- and intramolecular pathways coexist with a preeminent role of H bonds with J(a) = -2.8 and J(b) = -10.4 cm( 1). Not only are these values very consistent with the extracted set of parameters (J(1), J(2) = -11.6, -3.0 cm(-1)) but the possibility to generate leading exchange coupling through weak bonds is evidenced by means of wave function-based calculations. PMID- 21618978 TI - Contrasting reactivity and cancer cell cytotoxicity of isoelectronic organometallic iridium(III) complexes. AB - Replacing the N,N-chelating ligand 2,2'-bipyridine (bpy) in the Ir(III) pentamethylcyclopentadienyl (Cp*) complex [(eta(5)-C(5)Me(5))Ir(bpy)Cl](+) (1) with the C,N-chelating ligand 2-phenylpyridine (phpy) to give [(eta(5) C(5)Me(5))Ir(phpy)Cl] (2) switches on cytotoxicity toward A2780 human ovarian cancer cells (IC(50) values of >100 MUM for 1 and 10.8 MUM for 2). Ir-Cl hydrolysis is rapid for both complexes (hydrolysis equilibrium reached in <5 min at 278 K). Complex 2 forms adducts with both 9-ethylguanine (9-EtG) and 9 methyladenine (9-MeA), but preferentially with 9-EtG when in competition (ca. 85% of total Ir after 24 h). The X-ray crystal structure of [(eta(5) C(5)Me(5))Ir(phpy)(9-EtG-N7)]NO(3).1.5CH(2)Cl(2) confirms N7 binding to guanine. Two-dimensional NMR spectra show that complex 2 binds to adenine mainly through N1, consistent with density functional theory (DFT) calculations. DFT calculations indicate an interaction between the nitrogen of the NH(2) group (9 MeA) and carbons from phpy in the adenine adduct of complex 2. Calculations show that the most stable geometry of the adduct [(eta(5)-C(5)Me(5))Ir(phpy)(9-EtG N7)](+) (3b) has the C6O of 9-EtG orientated toward the pyridine ring of phpy, and for [(eta(5)-C(5)Me(5))Ir(phpy)(9-MeA-N1)](+) (4(N1)a), the NH(2) group of 9 EtA is adjacent to the phenyl ring side of phpy. Complex 2 is more hydrophobic than complex 1, with log P values of 1.57 and -0.95, respectively. The strong nucleobase binding and high hydrophobicity of complex 2 probably contribute to its promising anticancer activity. PMID- 21618979 TI - Synthesis, characterization, and reactivity of the stable iron carbonyl complex [Fe(CO)(N4Py)](ClO4)2: photoactivated carbon monoxide release, growth inhibitory activity, and peptide ligation. AB - Photoactivated carbon monoxide (CO) release by the iron carbonyl complex [Fe(II)(CO)(N4Py)](ClO(4))(2) (1) is described. Compound 1 is a low-spin ferrous complex that is highly stable and soluble in aerobic aqueous solutions. CO release was studied by the substitution of MeCN for CO, which displays saturation kinetics, and by the transfer of CO to deoxymyoglobin, which is slow in the dark but fast upon irradiation with UV light (365 nm). Compound 1 is active against PC 3 prostate cancer cells and shows potent photoinduced cytotoxicity. In addition, the iron carbonyl complex was attached to a short peptide toward the goal of tissue or cell-specific delivery. PMID- 21618980 TI - The Etard reaction: a DFT study. AB - The mechanism of the Etard reaction of chromylchloride in toluene, discovered more than a century ago, has been investigated by DFT calculations (B3LYP/6 31G(d)). The formation of the experimentally observed product can be rationalized by multiple CH-abstraction reactions. PMID- 21618981 TI - Building custom polysaccharides in vitro with an efficient, broad-specificity xyloglucan glycosynthase and a fucosyltransferase. AB - The current drive for applications of biomass-derived compounds, for energy and advanced materials, has led to a resurgence of interest in the manipulation of plant polymers. The xyloglucans, a family of structurally complex plant polysaccharides, have attracted significant interest due to their intrinsic high affinity for cellulose, both in muro and in technical applications. Moreover, current cell wall models are limited by the lack of detailed structure-property relationships of xyloglucans, due to a lack of molecules with well-defined branching patterns. Here, we have developed a new, broad-specificity "xyloglucan glycosynthase", selected from active-site mutants of a bacterial endoxyloglucanase, which catalyzed the synthesis of high molar mass polysaccharides, with complex side-chain structures, from suitable glycosyl fluoride donor substrates. The product range was further extended by combination with an Arabidopsis thaliana alpha(1->2)-fucosyltransferase to achieve the in vitro synthesis of fucosylated xyloglucans typical of dicot primary cell walls. These enzymes thus comprise a toolkit for the controlled enzymatic synthesis of xyloglucans that are otherwise impossible to obtain from native sources. Moreover, this study demonstrates the validity of a chemo-enzymatic approach to polysaccharide synthesis, in which the simplicity and economy of glycosynthase technology is harnessed together with the exquisite specificity of glycosyltransferases to control molecular complexity. PMID- 21618982 TI - Plasmon-based free-radical photopolymerization: effect of diffusion on nanolithography processes. AB - This Article interrogates the mechanisms responsible for nanoscale photopolymerization induced by confined and enhanced electromagnetic fields. Surface plasmon dipolar resonance of individual Ag nanoparticles was used as an optical near-field source to locally trigger the reaction of a photopolymerizable formulation. Laser excitation of the nanoparticles embedded in the formulation reproducibly generates polymer features with typical dimensions ranging from 2 nm to a few tens of nanometer. We have determined the physicochemical parameters and mechanisms controlling the spatial extent of the photopolymerization process. We found that the diffusion of the dye is the main process limiting the polymerization reaction, as opposed to what is observed at the microscale with an equivalent chemical system. This approach demonstrates that plasmon-based polymerization can achieve true nanometer scale resolution and also provides a unique opportunity to investigate photochemistry at this length scale. PMID- 21618983 TI - Redox properties of manganese-containing zirconia solid solution catalysts analyzed by in situ UV-vis spectroscopy and crystal field theory. AB - The optical absorption spectra of manganese-promoted sulfated zirconia, a highly active alkane isomerization catalyst, were found to be characterized by oxygen-to manganese charge-transfer transitions at 300-320 nm and d-d transitions of manganese ions at 580 and 680 nm. The latter were attributed to Mn(4+) and Mn(3+) ions, which are known to be incorporated in the zirconia lattice. The oxygen surroundings of these ions were modeled assuming a substitutional solid solution. The crystal field splittings, vibronic coupling constants, and oscillator strengths of the manganese ions were calculated on the basis of a cluster model that considers the manganese center as a complex with the adjacent ions of the lattice as ligands. The ratio of Mn(3+) to Mn(4+) ions was determined using the spectra and the model, and the relative concentrations of Mn(2+), Mn(3+), and Mn(4+) ions were determined with the help of the average valence known from X-ray absorption data in the literature. The redox behavior of manganese-promoted sulfated zirconia in oxidizing and inert atmosphere was elucidated at temperatures ranging from 323 to 773 K. PMID- 21618984 TI - Structural analysis of a prokaryotic ribosome using a novel amidinating cross linker and mass spectrometry. AB - The structure of the Escherichia coli ribosome, a 2.5 MDa ribonucleoprotein complex containing more than 50 proteins, was probed using the novel amidinating cross-linker diethyl suberthioimidate (DEST) and mass spectrometry. Peptide cross links derived from this complex structure were identified at high confidence (FDR 0.8%) from precursor mass measurements and collision-induced dissociation (CID) fragmentation spectra. The acquired cross-linking data were found to be in excellent agreement with the crystal structure of the E. coli ribosome. DEST cross-links are particularly amenable to strong cation exchange (SCX) chromatography, facilitating a large-scale analysis. SCX enrichment and fractionation were shown to increase the number of cross-link spectra matches in our analysis 10-fold. Evidence is presented that these techniques can be used to study complex interactomes. PMID- 21618985 TI - Paradynamics: an effective and reliable model for ab initio QM/MM free-energy calculations and related tasks. AB - Recent years have seen tremendous effort in the development of approaches with which to obtain quantum mechanics/molecular mechanics (QM/MM) free energies for reactions in the condensed phase. Nevertheless, there remain significant challenges to address, particularly, the high computational cost involved in performing proper configurational sampling and, in particular, in obtaining ab initio QM/MM (QM(ai)/MM) free-energy surfaces. One increasingly popular approach that seems to offer an ideal way to progress in this direction is the elegant metadynamics (MTD) approach. However, in the current work, we point out the subtle efficiency problems associated with this approach and illustrate that we have at hand what is arguably a more powerful approach. More specifically, we demonstrate the effectiveness of an updated version of our original idea of using a classical reference potential for QM(ai)/MM calculations [J. Phys. Chem. 1995, 99, 17516)], which we refer to as paradynamics (PD). This approach is based on the use of an empirical valence bond (EVB) reference potential, which is already similar to the real ab initio potential. The reference potential is fitted to the ab initio potential by an iterative and, to a great degree, automated refinement procedure. The corresponding free-energy profile is then constructed using the refined EVB potential, and the linear response approximation (LRA) is used to evaluate the QM(ai)/MM activation free-energy barrier. The automated refinement of the EVB surface (and thus the reduction of the difference between the reference and ab initio potentials) is a key factor in accelerating the convergence of the LRA approach. We apply our PD approach to a test reaction, namely, the S(N)2 reaction between a chloride ion and methyl chloride, and demonstrate that, at present, this approach is far more powerful and cost effective than the metadynamics approach (at least in its current implementation). We also discuss the general features of the PD approach in terms of its ability to explore complex systems and clarify that it is not a specialized approach limited to only accelerating QM(ai)/MM calculations with proper sampling, but rather can be used in a wide variety of applications. In fact, we point out that the use of a reference (CG) potential coupled with its PD refinement, as well as our renormalization approach, provides very general and powerful strategies that can be used very effectively to explore any property that has been studied by the MTD approach. PMID- 21618986 TI - Discovery of N-(1-ethylpropyl)-[3-methoxy-5-(2-methoxy-4-trifluoromethoxyphenyl) 6-methyl-pyrazin-2-yl]amine 59 (NGD 98-2): an orally active corticotropin releasing factor-1 (CRF-1) receptor antagonist. AB - The design, synthesis, and structure-activity relationships of a novel series of pyrazines, acting as corticotropin releasing factor-1 (CRF-1) receptor antagonists, are described. Synthetic methodologies were developed to prepare a number of substituted pyrazine cores utilizing regioselective halogenation and chemoselective derivatization. Noteworthy, an efficient 5-step synthesis was developed for the lead compound 59 (NGD 98-2), which required no chromatography. Compound 59 was characterized as an orally bioavailable, brain penetrant, and highly selective CRF-1 receptor antagonist. Occupancy of rat brain CRF-1 receptors was quantified using ex vivo receptor occupancy assays, using both brain tissue homogenates as well as brain slices receptor autoradiography. Behaviorally, oral administration of 59 significantly antagonized CRF-induced locomotor activity at doses as low as 10 mg/kg and dose-dependently reduced the restraint stress-induced ACTH increases. PMID- 21618988 TI - A theoretical study of the H-abstraction reactions from HOI by moist air radiolytic products (H, OH, and O (3P)) and iodine atoms (2P(3/2)). AB - The rate constants of the reactions of HOI molecules with H, OH, O ((3)P), and I ((2)P(3/2)) atoms have been estimated over the temperature range 300-2500 K using four different levels of theory. Geometry optimizations and vibrational frequency calculations are performed using MP2 methods combined with two basis sets (cc pVTZ and 6-311G(d,p)). Single-point energy calculations are performed with the highly correlated ab initio coupled cluster method in the space of single, double, and triple (pertubatively) electron excitations CCSD(T) using the cc pVTZ, cc-pVQZ, 6-311+G(3df,2p), and 6-311++G(3df,3pd) basis sets. Reaction enthalpies at 0 K were calculated at the CCSD(T)/cc-pVnZ//MP2/cc-pVTZ (n = T and Q), CCSD(T)/6-311+G(3df,2p)//MP2/6-311G(d,p), and CCSD(T)/6 311++G(3df,3pd)//MP2/6-311G(d,p) levels of theory and compared to the experimental values taken from the literature. Canonical transition-state theory with an Eckart tunneling correction is used to predict the rate constants as a function of temperature. The computational procedure has been used to predict rate constants for H-abstraction elementary reactions because there are actually no literature data to which the calculated rate constants can be directly compared. The final objective is to implement kinetics of gaseous reactions in the ASTEC (accident source term evaluation code) program to improve speciation of fission products, which can be transported along the reactor coolant system (RCS) of a pressurized water reactor (PWR) in the case of a severe accident. PMID- 21618987 TI - Coarse-grained model for PEGylated lipids: effect of PEGylation on the size and shape of self-assembled structures. AB - Self-assembly of polyethylene glycol (PEG)-grafted lipids at different sizes and concentrations was simulated using the MARTINI coarse-grained (CG) force field. The interactions between CG PEG and CG dipalmitoylglycerophosphocholine (DPPC) lipids were parametrized by matching densities of 19-mers of PEG and polyethylene oxide (PEO) grafted to the bilayer from all-atom simulations. Mixtures of lipids and PEG-grafted (M(w) = 550, 1250, and 2000) lipids in water self-assembled to liposomes, bicelles, and micelles at different ratios of lipids and PEGylated lipids. Average aggregate sizes decrease with increasing PEGylated-lipid concentration, in qualitative agreement with experiment. PEGylated lipids concentrate at the rims of bicelles, rather than at the planar surfaces; this also agrees with experiment, though the degree of segregation is less than that assumed in previous modeling of the experimental data. Charged lipids without PEG evenly distribute at the rim and planar surfaces of the bicelle. The average end to-end distances of the PEG on the PEGylated lipids are comparable in liposomes, bicelles (edge or planar surface), and micelles and only slightly larger than for an isolated PEG in solution. The ability of PEGylated lipids to induce the membrane curvature by the bulky headgroup with larger PEG, and thereby modulate the phase behavior and size of lipid assemblies, arises from their relative concentration. PMID- 21618989 TI - Synthesis and reactivity of unique heterocyclic structures en route to substituted diamines. AB - Rhodium-catalyzed oxidative cyclization of allylic hydroxylamine-derived sulfamate esters furnishes a novel family of bicyclic aziridines that serve as functional precursors to substituted diamines. Investigations with the N4-Troc form of these heterocycles have led to manifold improvements in reaction performance and scope and have revealed unique differences in the stability and reactivity of such compounds dictated by the choice of N4-protecting group. PMID- 21618990 TI - Simple N-sulfinyl-based chiral sulfur-olefin ligands for rhodium-catalyzed asymmetric 1,4-additions. AB - A variety of N-sulfinyl-based chiral sulfur-olefin ligands has been successfully developed for the first time for rhodium-catalyzed highly efficient and enantioselective 1,4-additions. The ease of synthesis and needless consideration of the carbon chirality makes this novel type of ligands attractive for asymmetric catalysis. PMID- 21618991 TI - Structural characterization of two alternate conformations in a calbindin D9k based molecular switch. AB - We have demonstrated that calbindin D(9k) can be converted into a calcium-sensing switch (calbindin-AFF) by duplicating the C-terminal half of the protein (residues 44-75) and appending it to the N-terminus (creating residues 44'-75'). This re-engineering results in a ligand-driven interconversion between two native folds: the wild-type structure (N) and a circularly permuted form (N'). The switch between N and N' is predicted to involve exchange of the 44-75 and 44'-75' segments, possibly linked to their respective folding and unfolding. Here we present direct structural evidence supporting the existence of N and N'. To isolate the N' and N conformations, we introduced the knockdown Ca(2+) binding mutation Glu -> Gln at position 65 (E65Q mutant) or at the analogous position 65' (E65'Q mutant). E65Q and E65'Q are therefore expected to adopt conformations N' and N, respectively, in the presence of calcium. Though the amino acid sequences of E65Q and E65'Q differ at only these two positions, nuclear magnetic resonance resonance assignments, chemical shifts, and paramagnetic relaxation enhancement data reveal that they take on separate structures when bound to calcium. Both proteins are comprised of a well-folded domain and a disordered region. However, the segment that is disordered in E65Q (residues 44-75) is folded in E65'Q, and the region that is disordered in E65'Q (residues 44'-75') is structured in E65Q. The results demonstrate that the N' N' conformational change is mediated by a mutually exclusive folding reaction in which folding of one segment of the protein is coupled to unfolding of another segment, and vice versa. PMID- 21618992 TI - Field emission from atomically thin edges of reduced graphene oxide. AB - Point sources exhibit low threshold electron emission due to local field enhancement at the tip. The development and implementation of tip emitters have been hampered by the need to position them sufficiently apart to achieve field enhancement, limiting the number of emission sites and therefore the overall current. Here we report low threshold field (< 0.1 V/MUm) emission of multiple electron beams from atomically thin edges of reduced graphene oxide (rGO). Field emission microscopy measurements show evidence for interference from emission sites that are separated by a few nanometers, suggesting that the emitted electron beams are coherent. On the basis of our high-resolution transmission electron microscopy, infrared spectroscopy, and simulation results, field emission from the rGO edge is attributed to a stable and unique aggregation of oxygen groups in the form of cyclic edge ethers. Such closely spaced electron beams from rGO offer prospects for novel applications and understanding the physics of linear electron sources. PMID- 21618993 TI - Recombinant protein hydrazides: application to site-specific protein PEGylation. AB - Here, we describe a novel method for the site-specific C-terminal PEGylation of recombinant proteins. This general approach exploits chemical cleavage of precursor intein-fusion proteins with hydrazine to directly produce recombinant protein hydrazides. This unique functionality within the protein sequence then facilitates site-specific C-terminal modification by hydrazone-forming ligation reactions. This approach was used to generate folded, site-specifically C terminal PEGylated IFNalpha2b and IFNbeta1b, which retained excellent antiviral activity, demonstrating the utility of this technology in the PEGylation of therapeutic proteins. As this methodology is straightforward to perform, is compatible with disulfide bonds, and is exclusively selective for the protein C terminus, it shows great potential as general technology for the site-specific engineering and labeling of recombinant proteins. PMID- 21618994 TI - [NiFe] and [FeS] cofactors in the membrane-bound hydrogenase of Ralstonia eutropha investigated by X-ray absorption spectroscopy: insights into O(2) tolerant H(2) cleavage. AB - Molecular features that allow certain [NiFe] hydrogenases to catalyze the conversion of molecular hydrogen (H(2)) in the presence of dioxygen (O(2)) were investigated. Using X-ray absorption spectroscopy (XAS), we compared the [NiFe] active site and FeS clusters in the O(2)-tolerant membrane-bound hydrogenase (MBH) of Ralstonia eutropha and the O(2)-sensitive periplasmic hydrogenase (PH) of Desulfovibrio gigas. Fe-XAS indicated an unusual complement of iron-sulfur centers in the MBH, likely based on a specific structure of the FeS cluster proximal to the active site. This cluster is a [4Fe4S] cubane in PH. For MBH, it comprises less than ~2.7 A Fe-Fe distances and additional longer vectors of >=3.4 A, consistent with an Fe trimer with a more isolated Fe ion. Ni-XAS indicated a similar architecture of the [NiFe] site in MBH and PH, featuring Ni coordination by four thiolates of conserved cysteines, i.e., in the fully reduced state (Ni SR). For oxidized states, short Ni-MUO bonds due to Ni-Fe bridging oxygen species were detected in the Ni-B state of the MBH and in the Ni-A state of the PH. Furthermore, a bridging sulfenate (CysSO) is suggested for an inactive state (Ni(ia)-S) of the MBH. We propose that the O(2) tolerance of the MBH is mainly based on a dedicated electron donation from a modified proximal FeS cluster to the active site, which may favor formation of the rapidly reactivated Ni-B state instead of the slowly reactivated Ni-A state. Thereby, the catalytic activity of the MBH is facilitated in the presence of both H(2) and O(2). PMID- 21618995 TI - Simulation of chaperonin effect on protein folding: a shift from nucleation condensation to framework mechanism. AB - The iterative annealing mechanism (IAM) of chaperonin-assisted protein folding is explored in a framework of a well-established coarse-grained protein modeling tool, which enables the study of protein dynamics in a time-scale well beyond classical all-atom molecular mechanics. The chaperonin mechanism of action is simulated for two paradigm systems of protein folding, B domain of protein A (BdpA) and B1 domain of protein G (GB1), and compared to chaperonin-free simulations presented here for BdpA and recently published for GB1. The prediction of the BdpA transition state ensemble (TSE) is in perfect agreement with experimental findings. It is shown that periodic distortion of the polypeptide chains by hydrophobic chaperonin interactions can promote rapid folding and leads to a decrease in folding temperature. It is also demonstrated how chaperonin action prevents kinetically trapped conformations and modulates the observed folding mechanisms from nucleation-condensation to a more framework like. PMID- 21618996 TI - Synthesis of new vitamin K analogues as steroid and xenobiotic receptor (SXR) agonists: insights into the biological role of the side chain part of vitamin K. AB - Vitamin K(2) has been demonstrated to induce gene expression related to bone formation through a nuclear steroid and xenobiotic receptor (SXR). We synthesized new vitamin K analogues with the same isoprene side chains symmetrically introduced at the 2 and 3 positions of 1,4-naphthoquinone and evaluated the transcriptional activity of the target gene. The transcriptional activity was related to the length of the side chain which allowed optimal interaction with ligand-binding domain of SXR. PMID- 21618997 TI - Discrimination of mononuclear and dinuclear dinitrosyl iron complexes (DNICs) by S K-edge X-ray absorption spectroscopy: insight into the electronic structure and reactivity of DNICs. AB - In addition to probing the formation of dinitrosyl iron complexes (DNICs) by the characteristic Fe K-edge pre-edge absorption energy ranging from 7113.4 to 7113.8 eV, the distinct S K-edge pre-edge absorption energy and pattern can serve as an efficient tool to unambiguously characterize and discriminate mononuclear DNICs and dinuclear DNICs containing bridged-thiolate and bridged-sulfide ligands. The higher Fe-S bond covalency modulated by the stronger electron-donating thiolates promotes the Fe -> NO pi-electron back-donation to strengthen the Fe-NO bond and weaken the NO-release ability of the mononuclear DNICs, which is supported by the Raman nu(Fe-NO) stretching frequency. The Fe-S bond covalency of DNICs further rationalizes the binding preference of the {Fe(NO)(2)} motif toward thiolates following the trend of [SEt](-) > [SPh](-) > [SC(7)H(4)SN](-). The relative d manifold energy derived from S K-edge XAS as well as the Fe K-edge pre-edge energy reveals that the electronic structure of the {Fe(NO)(2)}(9) core of the mononuclear DNICs [(NO)(2)Fe(SR)(2)](-) is best described as {Fe(III)(NO( ))(2)}(9) compared to [{Fe(III)(NO(-))(2)}(9)-{Fe(III)(NO(-))(2)}(9)] for the dinuclear DNICs [Fe(2)(MU-SEt)(MU-S)(NO)(4)](-) and [Fe(2)(MU-S)(2)(NO)(4)](2-). PMID- 21618998 TI - Synthesis and characterization of iron(II) and ruthenium(II) hydrido hydrazine complexes. AB - Treatment of trans-[MHCl(dmpe)(2)] (M = Fe, Ru) with hydrazine afforded the hydrido hydrazine complexes cis- and trans-[MH(N(2)H(4))(dmpe)(2)](+) which have been characterized by NMR spectroscopy ((1)H, (31)P, and (15)N). Both cis and trans isomers of the Fe complex and the trans isomer of the Ru complex were characterized by X-ray crystallography. Reactions with acid and base afforded a range of N(2)H(x) complexes, including several unstable hydrido hydrazido complexes. PMID- 21619000 TI - Monte Carlo simulation of the gas-phase volumetric adsorption system: effects of dosing volume size, incremental dosing amount, pore shape and size, and temperature. AB - We model the volumetric method commonly used in the measurement of gas-phase adsorption isotherms by using Monte Carlo (MC) simulation to study slit pore adsorption in a finite volume. Although the method has been used for a very long time, modeling of the operation by a Monte Carlo scheme to account properly for the exchange of mass between the solid and the finite dosing volume has not been widely studied in the literature. This paper presents the MC simulation of the system composed of the solid subsystem and the gas phase surrounding it. We show that not only the size of the dosing volume and the incremental dosing amount but also the pore shape, pore size, and temperature have significant effects on the unstable region of the phase diagram, especially when the system is going through a first-order transition. This study extends and augments the recent work of Puibasset et al. (1) by showing that the shape of the adsorbent walls and the incremental dosing amount can affect the chemical potential in the adsorption system. PMID- 21619001 TI - Diffusion of organic dyes in ionic liquid and giant micron sized ionic liquid mixed micelle: fluorescence correlation spectroscopy. AB - Diffusion of organic dyes in neat room temperature ionic liquid (RTIL) and RTIL mixed micelle has been studied by fluorescence correlation spectroscopy (FCS). We have selected two RTILs, 3-pentyl-1-methyl imidazolium bromide ([C5C1Im][Br]) and the corresponding tetra-fluoroborate ([C5C1Im][BF(4)]). Diffusion coefficients (D(t)) of three organic dyes--DCM (neutral), C480 (neutral), and C343 (anionic)- in these RTILs are ~100 times slower compared to water. This indicates very high viscosity of the RTILs. In contrast to water, the D(t) in RTIL exhibits a wide distribution which suggests the presence of heterogeneity (nanoscale organization). The presence of ions in the RTILs markedly affects diffusion in the RTILs. D(t)'s of C480 (neutral) and C343 (anionic) are very similar in water but in RTILs the ionic dye C343 diffuses 1.7 times slower than neutral C480. This is attributed to the electrostatic force exerted by the ions in the RTILs. In the giant (~2-4 MUm) [C5C1Im][Br]-triblock copolymer (P123) mixed micelle D(t) of DCM, C480, and C343 are found to be 7, 15, and 7 MUm(2) s(-1), respectively. The results are compared with those in P123 micelle and gel. PMID- 21619002 TI - 15N solid-state NMR as a probe of flavin H-bonding. AB - Flavins mediate a wide variety of chemical reactions in biology. To learn how one cofactor can be made to execute different reactions in different enzymes, we are developing solid-state NMR (SSNMR) to probe the flavin electronic structure, via the (15)N chemical shift tensor principal values (delta(ii)). We find that SSNMR has superior responsiveness to H-bonds, compared to solution NMR. H-bonding to a model of the flavodoxin active site produced an increase of 10 ppm in the delta(11) of N5, although none of the H-bonds directly engage N5, and solution NMR detected only a 4 ppm increase in the isotropic chemical shift (delta(iso)). Moreover SSNMR responded differently to different H-bonding environments, as H bonding with water caused delta(11) to decrease by 6 ppm, whereas delta(iso) increased by less than 1 ppm. Our density functional theoretical (DFT) calculations reproduce the observations, validating the use of computed electronic structures to understand how H-bonds modulate the flavin's reactivity. PMID- 21619003 TI - Electrostatic damping functions and the penetration energy. AB - The use of damping functions to correct the multipole expansion at short-range is explored. Damping functions for the terms in the multipole expansion can be determined ab initio as a linear combination of analytic functions of the separation between sites, but there are additional short-range terms that have different angular dependence. The approach provides a detailed ab initio description of the penetration energy correction to the multipole expansion in an easily comprehensible form. PMID- 21619004 TI - Catalytic asymmetric C-H insertions of rhodium(II) azavinyl carbenes. AB - A highly efficient enantioselective C-H insertion of azavinyl carbenes into unactivated alkanes has been developed. These transition metal carbenes are directly generated from readily available and stable 1-sulfonyl-1,2,3-triazoles in the presence of chiral Rh(II) carboxylates and are used for C-H functionalization of alkanes to access a variety of beta-chiral sulfonamides. PMID- 21619005 TI - High-resolution chemical imaging through tissue with an X-ray scintillator sensor. AB - We describe a novel method for high-resolution chemical imaging on a surface embedded in tissue. The sensor surface consists of an X-ray scintillator film coated in a thin film loaded with chemical indicator dye. A narrow scanning X-ray beam is used to excite luminescence from X-ray scintillators located within the beam. This luminescence passes through the indicator film, and the spectrum is analyzed to measure chemical concentrations at that location. A pH sensor is demonstrated with a dynamic range between pH 6-9 and noise level of 0.05 pH units using methyl-red dyed pH paper. The location of the interface between two types of scintillator films is obtained with 0.30 mm spatial resolution even though the images are highly blurred by 10 mm of chicken breast. This work has important applications for detecting pH changes on surfaces of implanted medical devices. PMID- 21619006 TI - Electroacoustics of particles dispersed in polymer gel. AB - This study examines the electroacoustics of particles dispersed in polymer hydrogels, with the particle size either less than or greater than the gel mesh size. When the particles are smaller than the gel mesh size, their acoustic vibration is resisted by only the background water medium, and the measured dynamic electrophoretic mobility, MU(d) (obtained in terms of colloid vibration current, CVI), is the same as that in water. For the case of particles larger than the gel mesh size, MU(d) is decreased due to trapping, and the net decrease depends on the viscoelastic properties of the gel. The gel mesh size was varied by varying its cross-link density, with the latter being characterized as the storage modulus, G'. The dependence of mobility on G', for systems of a given particle size, and on particle size, for gels of a given G', are investigated. The measured mobility remains constant as G' is increased (i.e., mesh size is decreased) up to a value of approximately 300 Pa, beyond which it decreases. In the second set of measurements, the trapped particle size was increased in a gel medium of constant mesh size, with G' being approximately 100 Pa. In this case, the measured MU(d) is found to be effectively constant over the particle size range studied (14-120 nm); that is, it is independent of the degree of trapping as expressed by the ratio of the particle size to the mesh size. PMID- 21619007 TI - Toward a universal model to calculate the solvation thermodynamics of druglike molecules: the importance of new experimental databases. AB - We demonstrate that a new free energy functional in the integral equation theory of molecular liquids gives accurate calculations of hydration thermodynamics for druglike molecules. The functional provides an improved description of excluded volume effects by incorporating two free coefficients. When the values of these coefficients are obtained from experimental data for simple organic molecules, the hydration free energies of an external test set of druglike molecules can be calculated with an accuracy of about 1 kcal/mol. The 3D RISM/UC method proposed here is easily implemented using existing computational software and allows in silico screening of the solvation thermodynamics of potential pharmaceutical molecules at significantly lower computational expense than explicit solvent simulations. PMID- 21619008 TI - Thermodynamics of optoplasmonic heating in fluid-filled gold-nanoparticle-plated capillaries. AB - Dynamic and equilibrium thermal behavior of plasmon-heated gold/silica capillary nanocomposite during evaporative cooling by water or butanol is accurately described at centimeter length scales by continuum optoplasmonic thermodynamics for continuous-wave laser irradiation of 15-50 mW. Gold nanoparticles randomly distributed on the capillary via electroless plating exhibited a composite extinction cross section of 66.74 +/- 0.72% of the area of the laser spot, more than 2-fold larger than the physical cross-section of the AuNPs. The extinction cross-section of the AuNPs capillary was invariant for incident laser powers of 15-150 mW and was reduced slightly in the presence of butanol and water due to absorption peak-shifting to lower energies. Introducing composite thermal parameters into the optoplasmonic thermodynamic relation extended its ability to predict heat transfer to laser powers of 100 and 150 mW for water and butanol, respectively. Nonlinear behaviors such as exponential thermal profiles caused by limited thermal conductivity and film boiling are identified at higher laser powers and prevent further extension of the relation. Mathematical reduction of temperature and time variables of the mathematical description shows it accounts for all measured thermodynamic effects when the aforementioned nonlinear behaviors are not present. This confirms that extraordinary thermal transport observed in some nanocomposites are absent for AuNP/silica systems in the given ranges, which allows a macroscale, continuum approach to describe thermal transport. PMID- 21619009 TI - Enantioselective total synthesis of a natural iridoid. AB - The first total synthesis of 6-hydroxy-7-(hydroxymethyl)-4 methylenehexahydrocyclopenta[c]pyran-1(3H)-one has been accomplished. A key feature of the synthesis includes facile construction of the bicyclic lactone intermediate via intramolecular Pd(0)-catalyzed allylic alkylation and the efficient transformation of this intermediate into the iridoid skeleton employing silicon tethered radical cyclization. PMID- 21619010 TI - Identifying specific small-molecule interactions using electrospray ionization mass spectrometry. AB - A simple method for establishing whether complexes composed of small molecules detected by electrospray ionization mass spectrometry (ES-MS) originate from specific interactions in solution or nonspecific binding during the ES process is described. The technique, referred to as the nonspecific probe method, exploits the tendency of small molecules to bind nonspecifically to macromolecules during the ES process to establish the presence of specific noncovalent interactions. To implement the method, a macromolecule probe (P(NS)), which does not bind specifically to any of the components present in solution, is added prior to ES MS analysis. The existence of specific small-molecule complexes is determined from an analysis of the measured distributions of the small molecules bound nonspecifically to P(NS). The principal assumption on which this methodology is based is that nonspecific binding of small molecules and their complexes to P(NS) during ES is a statistical (random) process. A mathematical framework for establishing the presence of specific heterocomplexes is presented. The reliability of the method for distinguishing specific from nonspecific small molecule interactions is illustrated for peptide-antibiotic and metal ion-ligand interactions in water. PMID- 21619011 TI - Syntheses, structures, and magnetic properties of three one-dimensional end-to end azide/cyanate-bridged copper(II) compounds exhibiting ferromagnetic interaction: new type of solid state isomerism. AB - The work in this paper presents the syntheses, structures, and magnetic properties of three end-to-end (EE) azide/cyanate-bridged copper(II) compounds [Cu(II)L(1)(MU(1,3)-NCO)](n).2nH(2)O (1), [Cu(II)L(1)(MU(1,3)-N(3))](n).2nH(2)O (2), and [Cu(II)L(2)(MU(1,3)-N(3))](n) (3), where the ligands used to achieve these species, HL(1) and HL(2), are the tridentate Schiff base ligands obtained from [1 + 1] condensations of salicylaldehyde with 4-(2-aminoethyl)-morpholine and 3-methoxy salicylaldehyde with 1-(2-aminoethyl)-piperidine, respectively. Compounds 1 and 2 crystallize in the monoclinic P2(1)/c space group, while compound 3 crystallizes in the orthorhombic Pbca space group. The metal center in 1-3 is in all cases pentacoordinated. Three coordination positions of the metal center in 1, 2, or 3 are satisfied by the phenoxo oxygen atom, imine nitrogen atom, and morpholine (for 1 and 2) or piperidine (for 3) nitrogen atom of one deprotonated ligand, [L(1)](-) or [L(2)](-). The remaining two coordination positions are satisfied by two nitrogen atoms of two end-to-end bridging azide ligands for 2 and 3 and one nitrogen atom and one oxygen atom of two end-to-end bridging cyanate ligands for 1. The coordination geometry of the metal ion is distorted square pyramidal in which one EE azide/cyanate occupies the apical position. Variable-temperature (2-300 K) magnetic susceptibilities of 1-3 have been measured under magnetic fields of 0.05 (from 2 to 30 K) and 1.0 T (from 30 to 300 K). The simulation reveals a ferromagnetic interaction in all three compounds with J values of +0.19 +/- 0.01, +0.79 +/- 0.01, and +1.25 +/- 0.007 cm(-1) for 1, 2, and 3, respectively. Compound 1 is the sole example of a ferromagnetically coupled EE cyanate-bridged 1-D copper(II) system. In addition, a rare example of supramolecular isomerism and a nice example of magnetic isomerism have been observed and most interestingly a new type of solid state isomerism has emerged as a result of the comparison of the structure and magnetic properties of 2 with a previously published compound (2A) having the same composition and even the same crystal system and space group (New J. Chem.2001, 25, 1203-1207). PMID- 21619012 TI - Catalytic Ni(II) in reactions of SmI2: Sm(II)- or Ni(0)-based chemistry? AB - The addition of catalytic amounts of Ni(II) salts provide enhanced reactivity and selectivity in numerous reactions of SmI(2), but the mechanistic basis for their effect is unknown. We report spectroscopic and kinetic studies on the mechanistic role of catalytic Ni(II) in the samarium Barbier reaction. The mechanistic studies presented herein show that the samarium Barbier reaction containing catalytic amounts of Ni(II) salts is driven solely by the reduction of Ni(II) to Ni(0) in a rate-limiting step. Once formed, Ni(0) inserts into the alkyl halide bond through oxidative addition to produce an organonickel species. During the reaction, the formation of colloidal Ni(0) occurs concomitantly with Ni(0) oxidative addition as an unproductive process. Overall, this study shows that a reaction thought to be driven by the unique features of SmI(2) is in fact a result of known Ni(0) chemistry. PMID- 21619013 TI - Conformational stability from variable-temperature infrared spectra of xenon solutions, r0 structural parameters, and vibrational assignment of pyrrolidine. AB - The infrared spectra of gaseous and variable-temperature liquid xenon solutions of pyrrolidine have been recorded. The enthalpy difference has been determined to be 109 +/- 11 cm(-1) (1.30 +/- 0.13 kJ mol(-1)) with the envelope-equatorial conformer more stable than the twist form with 37 +/- 3% present at ambient temperature. Ab initio calculations utilizing various basis sets up to MP2(full)/aug-cc-pVTZ have been used to predict the conformational stabilities, energy at the equatorial-axial saddle point, and barriers to planarity. From previously reported microwave rotational constants along with MP2(full)/6 311+G(d,p) predicted structural values, adjusted r(0) parameters have been obtained for both conformers. Heavy atom distances (A) of equatorial[twist] conformer are as follows: N(1)-C(2) = 1.469(3)[1.476(3)], N(1)-C(3) = 1.469(3)[1.479(3)], C(2)-C(4) = 1.541(3)[1.556(3)], C(3)-C(5) = 1.541(3)[1.544(3)], C(4)-C(5) = 1.556(3)[1.543(3)]; and angles (deg)?N(1)C(2)C(4) = 102.5(5)[107.6(5)], ?N(1)C(3)C(5) = 102.5(5)[105.4(5)], ?C(2)C(4)C(5) = 104.3(5)[104.6(5)], ?C(3)C(5)C(4) = 104.3(5)[103.7(5)], ?C(2)N(1)C(3) = 104.1(5)[103.9(5)], tauC(2)C(4)C(5)C(3) = 0.0(5)[13.5(5)]. A complete vibrational assignment is proposed for both conformers. PMID- 21619014 TI - Preparation of solvent-free, pore-spanning lipid bilayers: modeling the low tension of plasma membranes. AB - Plasma membrane tension, produced by the underlying cytoskeleton, governs many dynamic processes such as fusion, blebbing, exo- and endocytosis, cell migration, and adhesion. Here, a new protocol is introduced to model this intricate and often overlooked aspect of the plasma membrane. Lipid bilayers spanning pores of 600 nm radius were prepared by adsorption and spreading of giant unilamellar vesicles (GUVs) on moderately hydrophilic porous substrates prepared by gold coating and subsequent self-assembly of a mercaptoethanol monolayer. Rupture of GUVs formed tens of micrometer sized pore-spanning membrane patches displaying low tension of sigma <= 3.5 mN m(-1) and lateral diffusion constants of about 8 MUm(2) s(-1). Site-specific force indentation experiments were performed to determine membrane tension as a function of lipid composition: for pure DOPC bilayers, a tension of 1.018 +/- 0.014 mN m(-1) was measured, which was increased by the addition of cholesterol to 3.50 +/- 0.15 mN m(-1). Compared to DOPC, POPC bilayers displayed a larger tension of 2.00 +/- 0.09 mN m(-1). Addition and subsequent partitioning of 2-propanol was shown to significantly reduce the membrane tension as a function of its concentration. PMID- 21619015 TI - Reactive layer-by-layer deposition of poly(ethylene imine) and a precursor of TiO2: influence of the sodium chloride concentration on the film growth, interaction with hexacyanoferrate anions, and particle distribution in the film. AB - Films prepared according to a layer-by-layer (LBL) manner find increasing importance in many applications such as coatings with dedicated optical or electronic properties, particularly when including nanomaterials. An alternative way to prepare such hybrid layer-by-layer coatings is to perform sol-gel chemistry in a layer-by-layer manner. In this article, we highlight the importance of the NaCl concentration as a parameter to control the growth as well as the properties of LBL films made from poly(ethylene imine) as the organic counterpart and titanium IV (bisammoniumlactato)dihydroxyde ([Ti(lac)(2)(OH)(2)](2-)) as the precursor of TiO(2). An increase in the sodium chloride concentration leads to the faster growth of the film and to a decrease in the number of hexacyanoferrate anions remaining in the film after a buffer rinse. This may be due to a progressive increase in the fraction of negatively charged TiO(2) as suggested by transmission electron microscopy. In the presence of 0.5 M NaCl, the fraction of TiO(2) is close to 60% in mass. As a surprising finding, the films produced from 0.15 M NaCl are not homogeneously filled with TiO(2) even if the film is produced in an LBL fashion. The increased concentration of TiO(2) at the film-solution interface could constitute a barrier for the incorporation of the negatively charged redox probe. PMID- 21619016 TI - Study of thioflavin-T immobilized in porous silicon and the effect of different organic vapors on the fluorescence lifetime. AB - Steady-state and time-resolved emission techniques have been employed to study the fluorescence properties of thioflavin-T (ThT) adsorbed on oxidized porous silicon (PSi) surfaces, with an average pore size of ~10 nm. We found that the average fluorescence decay time of ThT, when it is adsorbed on the PSi surface, is rather long, tau(av) = 1.3 ns. We attribute this relatively long emission lifetime to the effect of the immobilization of ThT on the PSi surface, which inhibit the rotation of the aniline with respect to the benzothiazole moieties of ThT. We also measured the fluorescence properties of ThT in PSi samples in equilibrium with vapors of several liquids, such as methanol, acetonitrile, and water. We found that the fluorescence intensity drops by a factor of 10, and the average decay time, measured by a time-correlated single-photon counting technique, decreases by a factor of 3. We explain these results in terms of liquid condensation of the vapors in the PSi pores, which leads to partial dissolution of the ThT molecules in the liquid pools. PMID- 21619017 TI - Wetting of biopolymer coatings: contact angle kinetics and image analysis investigation. AB - The surface wetting of five biopolymers, used as coating materials for a plastic film, was monitored over a span of 8 min by means of the optical contact angle technique. Because most of the total variation was observed to occur during the first 60 s, we decided to focus on this curtailed temporal window. Initial contact angle values (theta(0)) ranged from ~91 degrees for chitosan to ~30 degrees for pullulan. However, the water drop profile began to change immediately following drop deposition for all biocoatings, confirming that the concept of water contact angle equilibrium is not applicable to most biopolymers. First, a three-parameter decay equation [theta(t) = theta(0) exp(kt(n))] was fit to the experimental contact angle data to describe the kinetics of the contact angle change for each biocoating. Interestingly, the k constant correlated well with the contact angle evolution rate and the n exponent seemed to be somehow linked to the physicochemical phenomena underlying the overall kinetics process. Second, to achieve a reliable description of droplet evolution, the contact angle (CA) analysis was coupled with image analysis (IA) through a combined geometric/trigonometric approach. Absorption and spreading were the key factors governing the overall mechanism of surface wetting during the 60 s analysis, although the individual quantification of both phenomena demonstrated that spreading provided the largest contribution for all biopolymers, with the only exception of gelatin, which showed two quasi-equivalent and counterbalancing effects. The possible correlation between these two phenomena and the topography of the biopolymer surfaces are then discussed on the basis of atomic force microscopy analyses. PMID- 21619018 TI - Near-field dynamics of optical Yagi-Uda nanoantennas. AB - We present near-field measurements of optical Yagi-Uda nanoantennas that are used in receiving mode. The eigenmode imaging of amplitude and phase by apertureless scanning near-field optical microscopy allows us to investigate the dynamics of the local out-of-plane electric field components and to visualize the temporal evolution of this time-harmonic reception process. The antenna directionality manifests itself by the dependence of the local field enhancement at the feed element on the illumination direction. Simulations taking into account the substrate confirm our observation of the directionality. Our work demonstrates the possibility to characterize multielement nanoantennas by electromagnetic antenna near-field scanners. PMID- 21619019 TI - A carpet cloak for visible light. AB - We report an invisibility carpet cloak device, which is capable of making an object undetectable by visible light. The cloak is designed using quasi conformal mapping and is fabricated in a silicon nitride waveguide on a specially developed nanoporous silicon oxide substrate with a very low refractive index (n<1.25). The spatial index variation is realized by etching holes of various sizes in the nitride layer at deep subwavelength scale creating a local effective medium index. The fabricated device demonstrates wideband invisibility throughout the visible spectrum with low loss. This silicon nitride on low index substrate can also be a general scheme for implementation of transformation optical devices at visible frequencies. PMID- 21619020 TI - Solution-processed sintered nanocrystal solar cells via layer-by-layer assembly. AB - Solar cells made by high temperature and vacuum processes from inorganic semiconductors are at a perceived cost disadvantage when compared with solution processed systems such as organic and dye-sensitized solar cells. We demonstrate that totally solution processable solar cells can be fabricated from inorganic nanocrystal inks in air at temperature as low as 300 degrees C. Focusing on a CdTe/ZnO thin-film system, we report solar cells that achieve power conversion efficiencies of 6.9% with greater than 90% internal quantum efficiency. In our approach, nanocrystals are deposited from solution in a layer-by-layer process. Chemical and thermal treatments between layers induce large scale grain formation, turning the 4 nm CdTe particles into pinhole-free films with an optimized average crystallite size of ~70 nm. Through capacitance-voltage measurements we demonstrate that the CdTe layer is fully depleted which enables the charge carrier collection to be maximized. PMID- 21619021 TI - Fluorescence properties of hydrophilic semiconductor nanoparticles with tridentate polyethylene oxide ligands. AB - In this contribution a facile, one-step synthesis of tridentate thiol functionalized PEO ligands and their ability to stabilize CdSe/CdS/ZnS core-shell shell nanoparticles in aqueous media are described. The PEO-coated quantum dots show colloidal stability as well as preserved fluorescence even at very low concentrations of a few nM. For improved ligand attachment and enhanced fluorescence properties a method for ligand exchange was developed, which includes formation of a ligand zinc complex before the actual exchange reaction. The stability and fluorescence properties in various aqueous buffers and cell media and at pH values down to pH 3 were investigated. The firm binding of the tridentate ligands to the particle surface makes this ligand-particle system a promising tool for biological applications. In addition, activation of the ligands' terminal hydroxyl group for covalent biofunctionalization by esterification with succinic acid is reported. PMID- 21619022 TI - Copper-catalyzed direct oxidative C-H amination of benzoxazoles with formamides or secondary amines under mild conditions. AB - A facile, efficient, and practical method for copper-catalyzed direct C-H amination of benzoxazoles with formamides or secondary amines has been developed. The system can be performed in the absence of external base and only requires O(2) or even air as oxidant. A variety of substituted benzoxazol-2-amines were synthesized with moderate to excellent yield. PMID- 21619023 TI - Mold design rules for residual layer-free patterning in thermal imprint lithography. AB - We present the mold design rules for assuring residual layer-free patterning in thermal imprint processes. Using simple relations for mass balance, structural stability, and work of adhesion, we derive the conditions with respect to the given single or multigeometrical feature of the mold, which are compared with simple thermal imprint experiments using soft imprint molds. Our analysis could serve as a guideline for designing the optimum mold geometry and selecting mold material in residual layer-free thermal imprint processes. PMID- 21619024 TI - Controlled growth of nanoparticles from solution with in situ liquid transmission electron microscopy. AB - Direct visualization of lead sulfide nanoparticle growth is demonstrated by selectively decomposing a chemical precursor from a multicomponent solution using in situ liquid transmission electron microscopy. We demonstrate reproducible control over growth mechanisms that dictate the final morphology of nanostructures while observing growth in real-time with subnanometer spatial resolution. Furthermore, while an intense electron beam can initiate nanoparticle growth, it is also shown that a laser can trigger the reaction independently of the imaging electrons. PMID- 21619025 TI - Synthesis and characterization of conformationally preorganized, (R)-diethylene glycol-containing gamma-peptide nucleic acids with superior hybridization properties and water solubility. AB - Developed in the early 1990s, peptide nucleic acid (PNA) has emerged as a promising class of nucleic acid mimic because of its strong binding affinity and sequence selectivity toward DNA and RNA and resistance to enzymatic degradation by proteases and nucleases; however, the main drawbacks, as compared to other classes of oligonucleotides, are water solubility and biocompatibility. Herein we show that installation of a relatively small, hydrophilic (R)-diethylene glycol ("miniPEG", R-MP) unit at the gamma-backbone transforms a randomly folded PNA into a right-handed helix. Synthesis of optically pure (R-MP)gammaPNA monomers is described, which can be accomplished in a few simple steps from a commercially available and relatively cheap Boc-l-serine. Once synthesized, (R-MP)gammaPNA oligomers are preorganized into a right-handed helix, hybridize to DNA and RNA with greater affinity and sequence selectivity, and are more water soluble and less aggregating than the parental PNA oligomers. The results presented herein have important implications for the future design and application of PNA in biology, biotechnology, and medicine, as well as in other disciplines, including drug discovery and molecular engineering. PMID- 21619026 TI - Synthesis of 5-iodopyrrolo[1,2-a]quinolines and indolo[1,2-a]quinolines via iodine-mediated electrophilic and regioselective 6-endo-dig ring closure. AB - The endo-cyclic ring closure of 1-(2-(substituted ethynyl)phenyl)-1H-pyrroles 3a t and 1-(2-(substituted ethynyl)phenyl)-H-indole 4a-o mediated by Lewis acid (I(2)) under mild conditions afforded substituted 5-iodopyrrolo[1,2-a]quinolines 5a-t and 5-iodoindolo[1,2-a]quinolines 6a-o in good to excellent yields. The reaction shows selective C-C bond formation on the more electrophilic alkynyl carbon, resulting in the regioselective 6-endo-dig-cyclized product. Iodo derivatives of pyrrolo- and indoloquinolines allow functional group diversification on the quinoline nucleus, which proves to be highly advantageous for structural and biological activity assessments. PMID- 21619027 TI - Synthesis and preliminary evaluation of pro-RNA 2'-O-masked with biolabile pivaloyloxymethyl groups in an RNA interference assay. AB - The cellular delivery of bioactive nucleic acid-based drugs such as small interfering RNA (siRNA) represents a major technical hurdle for their pharmaceutical application. Prodrug-like approaches provide an attractive concept to address the delivery problem. With the aim to prepare RNA-based prodrugs bearing biolabile protections which facilitate cellular uptake and are prone to be removed enzymatically inside cells in order to release functional RNA, we synthesized pro-RNA totally or partially masked in 2'-OH position with pivaloyloxymethyl (PivOM) groups. A suitable strategy has been developed to synthesize and to purify base-sensitive mixed 2'-OH/2'-O-PivOM oligoribonucleotides, and to include them in siRNA. In this strategy, the fluoride labile [(triisopropylsilyl)oxy]-benzyloxycarbonyl group (tboc) as nucleobase protection (for A and C), the TBS group as 2'-OH protection and the Q linker to solid-support were compatible with the PivOM groups masking some 2'-OH. We have taken advantage of the specific stability of the PivOM group to apply selected acidic, basic, and fluoride ions treatment for the deprotection and release of pro-RNA. This kind of pro-siRNA was studied in a human cell culture based RNAi assay and preliminary promising data are discussed. PMID- 21619028 TI - Enantioselective recognition of mandelic acid by a 3,6-dithiophen-2-yl-9H carbazole-based chiral fluorescent bisboronic acid sensor. AB - We have prepared chiral fluorescent bisboronic acid sensors with 3,6-dithiophen-2 yl-9H-carbazole as the fluorophore. The thiophene moiety was used to extend the pi-conjugation framework of the fluorophore in order to red-shift the fluorescence emission and, at the same time, to enhance the novel process where the fluorophore serves as the electron donor of the photoinduced electron transfer process (d-PET) of the boronic acid sensors; i.e., the background fluorescence of the sensor 1 at acidic pH is weaker compared to that at neutral or basic pH, in stark contrast to the typical a-PET boronic acid sensors (where the fluorophore serves as the electron acceptor of the photoinduced electron transfer process). The benefit of the d-PET boronic acid sensors is that the recognition of the hydroxylic acids can be achieved at acidic pH. We found that the thiophene moiety is an efficient pi-conjugation linker and electron donor; as a result, the d-PET contrast ratio of the sensors upon variation of the pH is improved 10-fold when compared to the previously reported d-PET sensors without the thiophene moiety. Enantioselective recognition of tartaric acid was achieved at acid pH, and the enantioselectivity (total response K(D)I(F)(D)/K(L)I(F)(L)) is 3.3. The fluorescence enhancement (I(F)(Sample)/I(F)(Blank)) of sensor 1 upon binding with tartaric acid is 3.5-fold at pH 3.0. With the fluorescent bisboronic acid sensor 1, enantioselective recognition of mandelic acid was achieved for the first time. To the best of our knowledge, this is the first time that the mandelic acid has been enantioselectively recognized using a chiral fluorescent boronic acid sensor. Chiral monoboronic acid sensor 2 and bisboronic acid sensor 3 without the thiophene moiety failed to enantioselectively recognize mandelic acid. Our findings with the thiophene-incorporated boronic acid sensors will be important for the design of d-PET fluorescent sensors for the enantioselective recognition of alpha-hydroxylic acids such as mandelic acid, given that it is currently a challenge to recognize these analytes with boronic acid fluorescent molecular sensors. PMID- 21619029 TI - Synthesis of 3-guaninyl- and 3-adeninyl-5-hydroxymethyl-2-pyrrolidinone nucleosides. AB - L- and D-glutamic acids, as well as trans-4-hydroxy-L-proline, are converted to the corresponding 3-guaninyl-5-hydroxymethyl-2-pyrrolidinone (4) or 3-adeninyl-5 hydroxymethyl-2-pyrrolidinone (5) nucleoside analog. The protecting group used to block the lactam nitrogen in key intermediates has a significant effect on the diastereoselectivity of the coupling reaction with adenine or guanine. PMID- 21619030 TI - Influence of histidine tag attachment on picosecond protein dynamics. AB - Polyhistidine affinity tags are routinely employed as a convenient means of purifying recombinantly expressed proteins. A tacit assumption is commonly made that His tags have little influence on protein structure and function. Attachment of a His tag to the N-terminus of the robust globular protein myoglobin leads to only minor changes to the electrostatic environment of the heme pocket, as evinced by the nearly unchanged Fourier transform infrared spectrum of CO bound to the heme of His-tagged myoglobin. Experiments employing two-dimensional infrared vibrational echo spectroscopy of the heme-bound CO, however, find that significant changes occur to the short time scale (picoseconds) dynamics of myoglobin as a result of His tag incorporation. The His tag mainly reduces the dynamics on the 1.4 ps time scale and also alters protein motions of myoglobin on the slower, >100 ps time scale, as demonstrated by the His tag's influence on the fluctuations of the CO vibrational frequency, which reports on protein structural dynamics. The results suggest that affinity tags may have effects on protein function and indicate that investigators of affinity-tagged proteins should take this into consideration when investigating the dynamics and other properties of such proteins. PMID- 21619031 TI - Unexpected nonadditivity effects in anion-pi complexes. AB - Several complexes of fluorine-substituted ethyne, ethene, butadiene, benzene, and [n]radialenes (n = 3-5) with two anions have been optimized at the RI-MP2/aug-cc pVTZ level of theory. The additivity of the anion-pi interaction was studied depending on the number of double bonds and fluorine atoms. Interesting nonadditivity effects were observed in the aromatic and antiaromatic complexes, which were analyzed by partitioning the total interaction energy into individual components, using Bader's theory of "atoms in molecules" and changes in the aromatic character of the ring upon complexation. PMID- 21619032 TI - Direct synthesis of spatially-controlled Pt-on-Pd bimetallic nanodendrites with superior electrocatalytic activity. AB - Here we report a facile synthesis of Pt-on-Pd bimetallic nanodendrites with a Pd interior and dendritic Pt exterior. The developed route rationally utilizes the spontaneous separation of the depositions of Pd and Pt, which endows direct formation of Pt-on-Pd nanodendrites. This is a truly simple and unique process that is quite different from the traditional seed-mediated growth strategy. Fine tuning of the Pt and Pd ratios afforded Pt-on-Pd nanodendrites with superior electrocatalytic activity in comparison with commercial Pt electrocatalysts. PMID- 21619034 TI - Comment on "HILIC-NMR: toward the identification of individual molecular components in dissolved organic matter". PMID- 21619035 TI - Effects of adsorbed arsenate on the rate of transformation of 2-line ferrihydrite at pH 10. AB - 2-Line ferrihydrite, a form of iron in uranium mine tailings, is a dominant adsorbent for elements of concern (EOC), such as arsenic. As ferrihydrite is unstable under oxic conditions and can undergo dissolution and subsequent transformation to hematite and goethite over time, the impact of transformation on the long-term stability of EOC within tailings is of importance from an environmental standpoint. Here, studies were undertaken to assess the rate of 2 line ferrihydrite transformation at varying As/Fe ratios (0.500-0.010) to simulate tailings conditions at the Deilmann Tailings Management Facility of Cameco Corporation, Canada. Kinetics were evaluated under relevant physical (~1 degrees C) and chemical conditions (pH ~10). As the As/Fe ratio increased from 0.010 to 0.018, the rate of ferrihydrite transformation decreased by 2 orders of magnitude. No transformation of ferrihydrite was observed at higher As/Fe ratios (0.050, 0.100, and 0.500). Arsenic was found to retard ferrihydrite dissolution and transformation as well as goethite formation. PMID- 21619036 TI - Ammonia activation by MU3-alkylidyne fragments supported on a titanium molecular oxide model. AB - Ammonolysis of the MU(3)-alkylidyne derivatives [{Ti(eta(5)-C(5)Me(5))(MU O)}(3)(MU(3)-CR)] [R = H (1), Me (2)] produces a trinuclear oxonitride species, [{Ti(eta(5)-C(5)Me(5))(MU-O)}(3)(MU(3)-N)] (3), via methane or ethane elimination, respectively. During the course of the reaction, the intermediates amido MU-alkylidene [{Ti(eta(5)-C(5)Me(5))(MU-O)}(3)(MU-CHR)(NH(2))] [(R = H (4), Me (5)] and MU-imido ethyl species [{Ti(eta(5)-C(5)Me(5))(MU-O)}(3)(MU-NH)Et] (6) were characterized and/or isolated. This achievement constitutes an example of characterization of the three steps of successive activation of N-H bonds in ammonia within the same transition-metal molecular system. The N-H sigma-bond activation of ammonia by the MU(3)-alkylidyne titanium species has been theoretically investigated by DFT method on [{Ti(eta(5)-C(5)H(5))(MU-O)}(3)(MU(3) CH)] model complex. The calculations complement the characterization of the intermediates, showing the multiple bond character of the terminal amido and the bridging nature of imido ligand. They also indicate that the sequential ammonia N H bonds activation process goes successively downhill in energy and occurs via direct hydron transfer to the alkylidyne group on organometallic oxides 1 and 2. The mechanism can be divided into three stages: (i) coordination of ammonia to a titanium center, in a trans disposition with respect to the alkylidyne group, and then the isomerization to adopt the cis arrangement, allowing the direct hydron migration to the MU(3)-alkylidyne group to yield the amido MU-alkylidene complexes 4 and 5, (ii) hydron migration from the amido moiety to the alkylidene group, and finally (iii) hydron migration from the MU-imido complex to the alkyl group to afford the oxo MU(3)-nitrido titanium complex 3 with alkane elimination. PMID- 21619038 TI - Magneto immunoassays for Plasmodium falciparum histidine-rich protein 2 related to malaria based on magnetic nanoparticles. AB - Magneto immunoassay-based strategies for the detection of Plasmodium falciparum histidine-rich protein 2 (HRP2) related to malaria are described for the first time by using magnetic micro- and nanoparticles. The covalent immobilization of a commercial monoclonal antibody toward the HRP2 protein in magnetic beads and nanoparticles was evaluated and compared. The immunological reaction for the protein HRP2 was successfully performed in a sandwich assay on magnetic micro- and nanoparticles by using a second monoclonal antibody labeled with the enzyme, horseradish peroxidase (HRP). Then, the modified magnetic particles were easily captured by a magneto sensor made of graphite-epoxy composite (m-GEC) which was also used as the transducer for the electrochemical detection. The performance of the immunoassay-based strategy with the electrochemical magneto immunosensors was successfully evaluated and compared with a novel magneto-ELISA based on optical detection using spiked serum samples. Improved sensitivity was obtained when using 300 nm magnetic nanoparticles in both cases. The electrochemical magneto immunosensor coupled with magnetic nanoparticles have shown better analytical performance in terms of limit of detection (0.36 ng mL(-1)), which is much lower than the LOD reported by other methods. Moreover, at a low level of HRP2 concentration of 31.0 ng mL(-1), a signal of 15.30 MUA was reached with a cutoff value of 0.34 MUA, giving a clear positive result with a non-specific adsorption ratio of 51. Due to the high sensitivity, this novel strategy offers great promise for rapid, simple, cost-effective, and on-site detection of falciparum malaria disease in patients, but also to screen out at-risk blood samples for prevention of transfusion-transmitted malaria. PMID- 21619039 TI - Energy and hole transfer between dyes attached to titania in cosensitized dye sensitized solar cells. AB - Cosensitization of broadly absorbing ruthenium metal complex dyes with highly absorptive near-infrared (NIR) organic dyes is a clear pathway to increase near infrared light harvesting in liquid-based dye-sensitized solar cells (DSCs). In cosensitized DSCs, dyes are intimately mixed, and intermolecular charge and energy transfer processes play an important role in device performance. Here, we demonstrate that an organic NIR dye incapable of hole regeneration is able to produce photocurrent via intermolecular energy transfer with an average excitation transfer efficiency of over 25% when cosensitized with a metal complex sensitizing dye (SD). We also show that intermolecular hole transfer from the SD to NIR dye is a competitive process with dye regeneration, reducing the internal quantum efficiency and the electron lifetime of the DSC. This work demonstrates the general feasibility of using energy transfer to boost light harvesting from 700 to 800 nm and also highlights a key challenge for developing highly efficient cosensitized dye-sensitized solar cells. PMID- 21619040 TI - Ice nucleation in emulsified aqueous solutions of antifreeze protein type III and poly(vinyl alcohol). AB - Antifreeze protein (AFP) III and poly(vinyl alcohol) (PVA) are known as anti-ice nucleating agents (anti-INAs), which inhibit heterogeneous ice nucleation. However, the effectiveness of these anti-INAs in inhibiting ice nucleation in water-in-oil (W/O) emulsions, in which homogeneous ice nucleation can be experimentally simulated, is unclear. In this study, the ice nucleation temperature in emulsified solutions of AFP III, PVA, and other nonanti-INA polymers was measured, and then the nucleation rate was analyzed based on classical nucleation theory. Results showed that ice nucleation was surface initiated and, except for PVA solutions, probably caused heterogeneously by the emulsifier, SPAN 65, at the droplet surfaces. In this nucleation mode, AFP III had no significant effect on the ice nucleation rate. In contrast, PVA exhibited ice-nucleating activity only at the droplet surfaces, suggesting that the nucleation is due to the interaction between PVA and SPAN 65. PMID- 21619041 TI - Spore-terminated cantilevers for chemical patterning on complex architectures. AB - Atomic force microscope tips terminated with spore cells are used to directly pattern onto glass and tissue surfaces. The spore cells act as sponges and eliminate the need to use microfabricated ink reservoirs during lithography. PMID- 21619042 TI - Molecular basis of the thermostability and thermophilicity of laminarinases: X ray structure of the hyperthermostable laminarinase from Rhodothermus marinus and molecular dynamics simulations. AB - Glycosyl hydrolases are enzymes capable of breaking the glycosidic linkage of polysaccharides and have considerable industrial and biotechnological applications. Driven by the later applications, it is frequently desirable that glycosyl hydrolases display stability and activity under extreme environment conditions, such as high temperatures and extreme pHs. Here, we present X-ray structure of the hyperthermophilic laminarinase from Rhodothermus marinus (RmLamR) determined at 1.95 A resolution and molecular dynamics simulation studies aimed to comprehend the molecular basis for the thermal stability of this class of enzymes. As most thermostable proteins, RmLamR contains a relatively large number of salt bridges, which are not randomly distributed on the structure. On the contrary, they form clusters interconnecting beta-sheets of the catalytic domain. Not all salt bridges, however, are beneficial for the protein thermostability: the existence of charge-charge interactions permeating the hydrophobic core of the enzymes actually contributes to destabilize the structure by facilitating water penetration into hydrophobic cavities, as can be seen in the case of mesophilic enzymes. Furthermore, we demonstrate that the mobility of the side-chains is perturbed differently in each class of enzymes. The side chains of loop residues surrounding the catalytic cleft in the mesophilic laminarinase gain mobility and obstruct the active site at high temperature. By contrast, thermophilic laminarinases preserve their active site flexibility, and the active-site cleft remains accessible for recognition of polysaccharide substrates even at high temperatures. The present results provide structural insights into the role played by salt-bridges and active site flexibility on protein thermal stability and may be relevant for other classes of proteins, particularly glycosyl hydrolases. PMID- 21619043 TI - Determining the conformational change that accompanies donor-acceptor distance fluctuations: an umbrella sampling analysis. AB - The response of a protein to variation of a specific coordinate can provide insights into the role of the overall architecture in the structural change. Given that the calculated potential of mean force governing the fluctuation of an electron transfer donor-acceptor distance in the NAD(P)H:Flavin oxidoreductase (Fre)/FAD complex was shown to agree with experiment, an analysis of the structural response of the rest of the protein to that distance change was made. Significant displacements are found throughout much of the protein, and the coupling pathway resulting in the structural changes was determined. A covariance analysis based on the quasiharmonic modes of the unperturbed protein was used to provide information concerning how the residue motions are correlated. It is found that, of the three regions identified as moving together in an NMR study, two undergo significant structural changes when the electron donor-acceptor distance is varied, and the third does not. PMID- 21619044 TI - Ni(II)/Zn catalyzed reductive coupling of aryl halides with diphenylphosphine oxide in water. AB - P-Arylation in water has been developed via cross-coupling of aryl halides with diphenylphosphine oxide (Ph(2)P(O)H) and (R(P))-(-)-menthyl benzylphosphinate catalyzed by NiCl(2).6H(2)O/Zn under relatively mild conditions. PMID- 21619045 TI - Isoflavone dimers and other bioactive constituents from the figs of Ficus mucuso. AB - Phytochemical investigation of the figs of Ficus mucuso led to the isolation of three new isoflavone dimer derivatives, mucusisoflavones A-C (1-3), together with 16 known compounds. Some of the isolates were tested in vitro for their inhibitory properties toward beta-glucuronidase and Plasmodium falciparum enoyl ACP reductase (PfENR) enzymes. Compound 1 (IC50) 0.68 MUM) showed inhibitory activity on beta-glucuronidase enzyme, while 3 (IC50) 7.69 MUM) exhibited a weak inhibitory activity against P. falciparum enoyl-ACP reductase (PfENR). PMID- 21619047 TI - A stereoselective oxidative polycyclization process mediated by a hypervalent iodine reagent. AB - Activation of phenol derivatives with a hypervalent iodine reagent promotes the formation of bicyclic and tricyclic products via a cationic cyclization process. The method allows efficient one-step syntheses of scaffolds present in several natural products and occurs with total stereocontrol, governed by 1,3 allylic strain interactions and by the configuration of the side chain double bonds. PMID- 21619048 TI - Performance characteristics of a new hybrid quadrupole time-of-flight tandem mass spectrometer (TripleTOF 5600). AB - The TripleTOF 5600 System, a hybrid quadrupole time-of-flight mass spectrometer, was evaluated to explore the key figures of merit in generating peptide and protein identifications that included spectral acquisition rates, data quality, proteome coverage, and biological depth. Employing a Saccharomyces cerevisiae tryptic digest, careful consideration of several performance features demonstrated that the speed of the TripleTOF contributed most to the resultant data. The TripleTOF system was operated with 8, 20, and 50 MS/MS events in an effort to compare with other MS technologies and to demonstrate the abilities of the instrument platform. PMID- 21619049 TI - Determination of toxicity equivalent factors for paralytic shellfish toxins by electrophysiological measurements in cultured neurons. AB - The establishment of toxicity equivalent factors to develop alternative methods to animal bioassays for marine-toxin detection is an urgent need in the field of phycotoxin research. Paralytic shellfish poisoning (PSP) is one of the most severe forms of food poisoning. The toxins responsible for this type of poisoning are highly toxic natural compounds produced by dinoflagellates, which bind to voltage-gated Na(+) channels causing the blockade of action potential propagation. In spite of the fact that several standards of PSP toxins are currently commercially available, there is scarcity of data on the biological activity of these toxins, a fact that limits the calculation of their toxicity equivalent factors. We have evaluated the potency of the commercial PSP toxin standards for their ability to inhibit voltage-dependent sodium currents in cultured neuronal cells by electrophysiological measurements. The in vitro potencies of the PSP toxin standards as indicated by their IC(50) values were in the order Neosaxitoxin (NeoSTX) > decarbamoylsaxitoxin (dcSTX) > saxitoxin (STX) > gonyautoxin 1,4 (GTX1,4) > decarbamoylneosaxitoxin (dcNeoSTX) > gonyautoxin 2,3 (GTX2,3) > decarbamoylgonyautoxin 2,3 (dcGTX2,3) > gonyautoxin 5 (GTX5) > N sulfocarbamoyl-gonyautoxin-2 and -3 (C1,2). The data obtained in this in vitro analysis correlated well with their previously reported toxicity values. PMID- 21619051 TI - Survivin silencing as a promising strategy to enhance the sensitivity of cancer cells to chemotherapeutic agents. AB - Since clinical application of conventional cancer therapies is usually limited by drug resistance and toxic side effects, combination of chemotherapeutic agents with gene therapy appears as an attractive therapeutic strategy to overcome these issues. Being selectively expressed in tumor tissues, survivin is a promising target for the development of anticancer strategies aimed at eliminating tumor cells while sparing normal tissues. In this work, we achieved substantial protein knockdown in a number of human cell lines, namely, A549, HeLa and MCF-7 cells which overexpress survivin, after treatment with anti-survivin siRNAs, which was associated with a significant reduction of cell viability, when compared to treatment with control siRNAs. Interestingly, when the survivin-silencing approach was combined with a chemotherapeutic agent, an enhancement of the therapeutic effect was achieved. Treatment with anti-survivin siRNAs resulted in high levels of caspase 3/7 activation, and an enhancement of this effect was observed when survivin silencing was combined with vinblastine. In addition, we showed that for A549 and HeLa cells survivin silencing contributes to the reversion of cell resistance to doxorubicin. Overall, we demonstrate that the combination of a survivin-directed silencing strategy with chemotherapeutic agents constitutes a valuable approach for cancer treatment. PMID- 21619050 TI - Regulatory mechanisms of tumor suppressor P16(INK4A) and their relevance to cancer. AB - P16(INK4A) (also known as P16 and MTS1), a protein consisting exclusively of four ankyrin repeats, is recognized as a tumor suppressor mainly because of the prevalence of genetic inactivation of the p16(INK4A) (or CDKN2A) gene in virtually all types of human cancers. However, it has also been shown that an elevated level of expression (upregulation) of P16 is involved in cellular senescence, aging, and cancer progression, indicating that the regulation of P16 is critical for its function. Here, we discuss the regulatory mechanisms of P16 function at the DNA level, the transcription level, and the posttranscriptional level, as well as their implications for the structure-function relationship of P16 and for human cancers. PMID- 21619052 TI - Use of urine volatile organic compounds to discriminate tuberculosis patients from healthy subjects. AB - Development of noninvasive methods for tuberculosis (TB) diagnosis, with the potential to be administered in field situations, remains as an unmet challenge. A wide array of molecules are present in urine and reflect the pathophysiological condition of a subject. With infection, an alteration in the molecular constituents is anticipated, characterization of which may form a basis for TB diagnosis. In the present study volatile organic compounds (VOCs) in human urine derived from TB patients and healthy controls were identified and quantified using headspace gas chromatography/mass spectrometry (GC/MS). We found significant (p < 0.05) increase in the abundance of o-xylene (6.37) and isopropyl acetate (2.07) and decreased level of 3-pentanol (0.59), dimethylstyrene (0.37), and cymol (0.42) in TB patients compared to controls. These markers could discriminate TB from healthy controls and related diseases like lung cancer and chronic obstructive pulmonary disorder. This study suggests a possibility of using urinary VOCs for the diagnosis of human TB. PMID- 21619053 TI - On the structure selectivity of clathrate hydrates. AB - We propose a strategic method to search for a new clathrate hydrate by investigating the selectivity of its crystalline structure, which has been expected to depend mostly on the guest species and less on the thermodynamic conditions. Relative stability among various morphologies is examined in terms of the chemical potential of water of clathrate hydrates in equilibrium with guest gases. This evaluation is performed by calculating the free energy of cage occupancy according to the standard statistical mechanical theory with the aid of the Yarmolyuk and Kripyakevich's rule on the Frank-Kasper type alloys to estimate the numbers of cages of various types. Thus, a comprehensive interpretation of the selectivity of crystalline structures is successfully made. We explain why two major structures are stable in most of the thermodynamic conditions and establish a relation between polymorphism of clathrate hydrate and the guest size and other parameters, thereby suggesting a way to find a new clathrate hydrate by appropriate choice of the guest species and/or the thermodynamic properties. It is found that there is a small room in the above parameter space for the other structure than the major two to be the most stable, including a new structure. In addition, simple but thorough elucidation is given for preferential formation of TS-I structure in bromine hydrate. PMID- 21619054 TI - Electronic packing frustration in complex intermetallic structures: the role of chemical pressure in Ca2Ag7. AB - The assignment of distinct roles to electronics and sterics has a long history in our rationalization of chemical phenomena. Exploratory synthesis in the field of intermetallic compounds challenges this dichotomy with a growing list of phases whose structural chemistry points to an interplay between atomic size effects and orbital interactions. In this paper, we begin with a simple model for how this interdependence may arise in the dense atomic packing of intermetallics: correlations between interatomic distances lead to the inability of a phase to optimize bonds without simultaneously shortening electronically under-supported contacts, a conflict we term electronic packing frustration (EPF). An anticipated consequence of this frustration is the emergence of chemical pressures (CPs) acting on the affected atoms. We develop a theoretical method based on DFT calibrated MU(2)-Huckel calculations for probing these CP effects. Applying this method to the Ca(2)Ag(7) structure, a variant of the CaCu(5) type with defect planes, reveals its formation is EPF-driven. The defect planes resolve severe CPs surrounding the Ca atoms in a hypothetical CaCu(5)-type CaAg(5) phase. CP analysis also points to a rationale for these results in terms of a CP analogue of the pressure-distance paradox and predicts that the impetus for defect plane insertion is tunable via variations in the electron count. PMID- 21619055 TI - Electron paramagnetic resonance and computational studies of radicals derived from boron-substituted N-heterocyclic carbene boranes. AB - Fifteen second-generation NHC-ligated boranes with aryl and alkyl substituents on boron were prepared, and their radical chemistry was explored by electron paramagnetic resonance (EPR) spectroscopy and calculations. Hydrogen atom abstraction from NHC-BH(2)Ar groups produced boryl radicals akin to diphenylmethyl with spin extensively delocalized across the NHC, BH, and aryl units. All of the NHC-B.HAr radicals studied abstracted Br-atoms from alkyl bromides. Radicals with bulky N,N'-dipp substituents underwent dimerization about 2 orders of magnitude more slowly than first-generation NHC-ligated trihydroborates. The evidence favored head-to-head coupling yielding ligated diboranes. The first ligated diboranyl radical, with a structure intermediate between that of ligated diboranes and diborenes, was spectroscopically characterized during photolysis of di-t-butyl peroxide with N,N'-di-t-butyl imidazol-2-ylidene phenylborane. The reactive site of B-alkyl-substituted NHC boranes switched from the boron center to the alkyl substituent for both linear and branched alkyl groups. The beta-borylalkyl radicals obtained from N,N'-dipp substituted boranes underwent exothermic beta-scissions with production of dipp Imd-BH(2). radicals and alkenes. The reverse additions of NHC-boryl radicals to alkenes are probably endothermic for alkyl-substituted alkenes, but exothermic for conjugated alkenes (addition of an NHC-boryl radical to 1,1-diphenylethene was observed). A cyclopropylboryl radical was observed, but, unlike other alpha cyclopropyl-substituted radicals, this showed no propensity for ring-opening. PMID- 21619056 TI - Development of a new high-performance liquid chromatography-electrospray ionization time-of-flight mass spectrometry method for the determination of low molecular mass organic acids in plant tissue extracts. AB - A liquid chromatography-electrospray ionization time-of-flight mass spectrometry method has been developed for the direct and simultaneous determination of 10 low molecular mass organic acids in different plant tissue extracts. The method does not include a derivatization step. Quantification was accomplished using (13)C labeled malic and succinic acids as internal standards. Good limits of detection (0.05-1.27 pmol) were obtained for malic, 2-oxoglutaric, succinic, quinic, shikimic, cis-aconitic, and citric acids, whereas for oxalic, ascorbic, and fumaric acids limits of detection were 255, 25, and 11 pmol, respectively. Repeatability values for the retention time and peak area were <5%, with the exception of ascorbic acid. Analyte recovery was between 92% and 110% in most cases, with the exception of oxalic (39-108%), 2-oxoglutaric (44-69%), and ascorbic (22-86%) acids, which may require specific extraction procedures and use of the corresponding (13)C-labeled organic acid as internal standards to improve accuracy. The method was applied to three types of plant materials: sugar beet leaf extracts, tomato xylem sap, and commercial orange juice. PMID- 21619057 TI - Preview: a program for surveying shotgun proteomics tandem mass spectrometry data. AB - Database search programs for peptide identification by tandem mass spectrometry ask their users to set various parameters, including precursor and fragment mass tolerances, digestion specificity, and allowed types of modifications. Even proteomics experts with detailed knowledge of their samples may find it difficult to make these choices without significant investigation, and poor choices can lead to missed identifications and misleading results. Here we describe a program called Preview that analyzes a set of mass spectra for mass errors, digestion specificity, and known and unknown modifications, thereby facilitating parameter selection. Moreover, Preview optionally recalibrates mass over charge measurements, leading to further improvement in identification results. In a study of Bruton's tyrosine kinase, we find that the use of Preview improved the number of confidently identified mass spectra and phosphorylation sites by about 50%. PMID- 21619058 TI - Infrared spectroscopy of Mn+ (H2O) and Mn2+ (H2O) via argon complex predissociation. AB - Singly and doubly charged manganese-water cations, and their mixed complexes with attached argon atoms, are produced by laser vaporization in a pulsed nozzle source. Complexes of the form Mn(+)(H(2)O)Ar(n) (n = 1-4) and Mn(2+)(H(2)O)Ar(4) are studied via mass-selected infrared photodissociation spectroscopy, detected in the mass channels corresponding to the elimination of argon. Sharp resonances are detected for all complexes in the region of the symmetric and asymmetric stretch vibrations of water. With the guidance of density functional theory computations, specific vibrational band resonances are assigned to complexes having different argon attachment configurations. In the small singly charged complexes, argon adds first to the metal ion site and later in larger clusters to the hydrogens of water. The doubly charged complex has argon only on the metal ion. Vibrations in all of these complexes are shifted to lower frequencies than those of the free water molecule. These shifts are greater when argon is attached to hydrogen and also greater for the dication compared to the singly charged species. Cation binding also causes the IR intensities for water vibrations to be much greater than those of the free water molecule, and the relative intensities are greater for the symmetric stretch than the asymmetric stretch. This latter effect is also enhanced for the dication complex. PMID- 21619059 TI - Nanospheres delivering the EGFR TKI AG1478 promote optic nerve regeneration: the role of size for intraocular drug delivery. AB - Promoting nerve regeneration involves not only modulating the postinjury microenvironment but also ensuring survival of injured neurons. Sustained delivery of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) has been shown to promote the survival and regeneration of neurons, but systemic administration is associated with significant side effects. We fabricated poly(lactic-co-glycolic acid) (PLGA) microspheres and nanospheres containing the EGFR TKI 4-(3-chloroanilino)-6,7-dimethoxyquinazoline (AG1478) for intravitreal administration in a rat optic nerve crush injury model. Upon administration, less backflow from the injection site was observed when injecting nanospheres compared to microspheres. Two weeks after intravitreal delivery, we were able to detect microspheres and nanospheres in the vitreous using coumarin-6 fluorescence, but fewer microspheres were observed compared to the nanospheres. At four weeks only nanospheres could be detected. AG1478 microspheres and nanospheres promoted optic nerve regeneration at two weeks, and at four weeks evidence of regeneration was found only in the nanosphere-injected animals. This observation could be attributed to the ease of administration of the nanospheres versus the microspheres, which in turn led to an increased amount of spheres delivered to the vitreous in the nanosphere group compared to the microsphere group. These data provide evidence for use of PLGA nanospheres to deliver AG1478 intravitreally in a single administration to promote nerve regeneration. PMID- 21619060 TI - Stable isotope labeling with amino acids in cell culture based mass spectrometry approach to detect transient protein interactions using substrate trapping. AB - The analysis of protein interactors in protein complexes can yield important insight into protein function and signal transduction. Thus, a reliable approach to distinguish true interactors from nonspecific interacting proteins is of utmost importance for accurate data interpretation. Although stringent purification methods are critical, challenges still remain in the selection of criteria that will permit the objective differentiation of true members of the protein complex from nonspecific background proteins. To address these challenges, we have developed a quantitative proteomic strategy combining stable isotope labeling with amino acids in cell culture (SILAC), affinity substrate trapping, and gel electrophoresis followed by liquid chromatography-tandem mass spectrometry (geLC-MS/MS) protein quantitation. ATP hydrolysis-deficient vacuolar protein sorting-associated protein 4B (Vps4B) was used as the "bait" protein which served as a substrate trap since its lack of ATP hydrolysis enzymatic activity allows the stabilization of its transiently associated interacting proteins. A significant advantage of our approach is the use of our new in-house developed software program for SILAC-based mass spectrometry quantitation, which further facilitates the differentiation between the bait protein, endogenous bait interacting proteins, and nonspecific binding proteins based on their protein ratios. The strategy presented herein is applicable to the analysis of other protein complexes whose compositions are dependent upon the ATP hydrolysis activity of the bait protein used in affinity purification studies. PMID- 21619061 TI - Conductance preservation of carbene-functionalized metallic single-walled carbon nanotubes. PMID- 21619062 TI - Folate-functionalized unimolecular micelles based on a degradable amphiphilic dendrimer-like star polymer for cancer cell-targeted drug delivery. AB - A folate-functionalized degradable amphiphilic dendrimer-like star polymer (FA DLSP) with a well-defined poly(L-lactide) (PLLA) star polymer core and six hydrophilic polyester dendrons based on 2,2-bis(hydroxymethyl) propionic acid was successfully synthesized to be used as a nanoscale carrier for cancer cell targeted drug delivery. This FA-DLSP hybrid formed unimolecular micelles in the aqueous solution with a mean particle size of ca. 15 nm as determined by dynamic light scattering and transmission electron microscopy. To study the feasibility of FA-DLSP micelles as a potential nanocarrier for targeted drug delivery, we encapsulated a hydrophobic anticancer drug, doxorubicin (DOX), in the hydrophobic core, and the loading content was determined by UV-vis analysis to be 4 wt %. The DOX-loaded FA-DLSP micelles demonstrated a sustained release of DOX due to the hydrophobic interaction between the polymer core and the drug molecules. The hydrolytic degradation in vitro was monitored by weight loss and proton nuclear magnetic resonance spectroscopy to gain insight into the degradation mechanism of the FA-DLSP micelles. It was found that the degradation was pH-dependent and started from the hydrophilic shell gradually to the hydrophobic core. Flow cytometry and confocal microscope studies revealed that the cellular binding of the FA-DLSP hybrid against human KB cells with overexpressed folate-receptors was about twice that of the neat DLSP (without FA). The in vitro cellular cytotoxicity indicated that the FA-DLSP micelles (without DOX) had good biocompatibility with KB cells, whereas DOX-loaded micelles exhibited a similar degree of cytotoxicity against KB cells as that of free DOX. These results clearly showed that the FA-DLSP unimolecular micelles could be a promising nanosize anticancer drug carrier with excellent targeting property. PMID- 21619064 TI - Artificial DNA lattice fabrication by noncomplementarity and geometrical incompatibility. AB - Fabrication of DNA nanostructures primarily follows two fundamental rules. First, DNA oligonucleotides mutually combine by Watson-Crick base-pairing rules between complementary base sequences. Second, the geometrical compatibility of the DNA oligonucleotide must match for lattices to form. Here we present a fabrication scheme of DNA nanostructures with noncomplementary and/or geometrically incompatible DNA oligonucleotides, which contradicts conventional DNA structure creation rules. Quantitative analyses of DNA lattice sizes were carried out to verify the unfavorable binding occurrences, which correspond to errors in algorithmic self-assembly. Further studies of these types of bindings may shed more light on the exact mechanisms at work in the self-assembly of DNA nanostructures. PMID- 21619063 TI - Facile synthetic route for surface-functionalized magnetic nanoparticles: cell labeling and magnetic resonance imaging studies. AB - Currently available methods to stably disperse iron oxide nanoparticles (IONPs) in aqueous solution need to be improved due to potential aggregation, reduction of superparamagnetism, and the use of toxic reagents. Herein, we present a facile strategy for aqueous transfer and dispersion of organic-synthesized IONPs using only polyethylene glycol (PEG), a biocompatible polymer. A library of PEG derivatives was screened, and it was determined that amine-functionalized six armed PEG, 6(PEG-NH(2)), was the most effective dispersion agent. The 6(PEG NH(2))-modified IONPs (IONP-6PEG) were stable after extensive washing, exhibited high superparamagnetism, and could be used as a platform material for secondary surface functionalization with bioactive polymers. IONP-6PEG biofunctionalized with hyaluronic acid (IONP-6PEG-HA) was shown to specifically label mesenchymal stem cells and demonstrate MR contrast potential with high r(2) relaxivity (442.7 s(-1)mM(-1)) compared to the commercially available Feridex (182.1 s(-1)mM(-1)). PMID- 21619065 TI - Model structures of Helicobacter pylori UreD(H) domains: a putative molecular recognition platform. AB - The analysis of the sequence of Helicobacter pylori UreD(H), an accessory protein involved in the activation of urease through the assembly of the Ni(2+) containing active site, revealed the presence of two domains. The structure of these domains was calculated using threading and modeling algorithms. A search for putative binding sites on the protein surface was carried out using dedicated algorithms sensitive to either sequence conservation or structural similarity based on geometry and physicochemical properties. The results suggest that UreD(H) acts as a multifunctional molecular recognition platform facilitating the interaction between apo-urease and the ancillary proteins UreG, UreF, and UreE, responsible for nickel trafficking and delivering. PMID- 21619066 TI - Selective kainate receptor (GluK1) ligands structurally based upon 1H cyclopentapyrimidin-2,4(1H,3H)-dione: synthesis, molecular modeling, and pharmacological and biostructural characterization. AB - The physiological function of kainate receptors (GluK1-GluK5) in the central nervous system is not fully understood yet. With the aim of developing potent and selective GluK1 ligands, we have synthesized a series of new thiophene-based GluK1 agonists (6a-c) and antagonists (7a-d). Pharmacological evaluation revealed that they are selective for the GluK1 subunit, with 7b being the most subtype selective ligand reported to date (GluK1 vs GluK3). The antagonist 7a was cocrystallized with the GluK1 ligand binding domain, and an X-ray crystallographic analysis revealed the largest flexibility in GluK1 ligand binding domain opening upon binding of a ligand seen to date. The results provide new insights into the molecular mechanism of GluK1 receptor ligand binding and pave the way to the development of new tool compounds for studying kainate receptor function. PMID- 21619067 TI - Reactivity of deoxy- and oxyferrous dehaloperoxidase B from Amphitrite ornata: identification of compound II and its ferrous-hydroperoxide precursor. AB - Dehaloperoxidase (DHP) from the terebellid polychaete Amphitrite ornata is a bifunctional enzyme that possesses both hemoglobin and peroxidase activities. The bifunctional nature of DHP as a globin peroxidase appears to be at odds with the traditional starting oxidation state for each individual activity. Namely, reversible oxygen binding is only mediated via a ferrous heme in globins, and peroxidase activity is initiated from ferric centers and to the exclusion of the oxyferrous oxidation state from the peroxidase cycle. Thus, to address what appears to be a paradox, herein we report the details of our investigations into the DHP catalytic cycle when initiated from the deoxy- and oxyferrous states using biochemical assays, stopped-flow UV-visible, and rapid-freeze-quench electron paramagnetic resonance spectroscopies, and anaerobic methods. We demonstrate the formation of Compound II directly from deoxyferrous DHP B upon its reaction with hydrogen peroxide and show that this occurs both in the presence and in the absence of trihalophenol. Prior to the formation of Compound II, we have identified a new species that we have preliminarily attributed to a ferrous-hydroperoxide precursor that undergoes heterolysis to generate the aforementioned ferryl intermediate. Taken together, the results demonstrate that the oxyferrous state in DHP is a peroxidase competent starting species, and an updated catalytic cycle for DHP is proposed in which the ferric oxidation state is not an obligatory starting point for the peroxidase catalytic cycle of dehaloperoxidase. The data presented herein provide a link between the peroxidase and oxygen transport activities, which furthers our understanding of how this bifunctional enzyme is able to unite its two inherent functions in one system. PMID- 21619068 TI - Luminescent trimethoprim-polyaminocarboxylate lanthanide complex conjugates for selective protein labeling and time-resolved bioassays. AB - Labeling proteins with long-lifetime emitting lanthanide (III) chelate reporters enables sensitive, time-resolved luminescence bioaffinity assays. Heterodimers of trimethoprim (TMP) covalently linked to various cs124-sensitized, polyaminocarboxylate chelates stably retain lanthanide ions and exhibit quantum yields of europium emission up to 20% in water. A time-resolved, luminescence resonance energy transfer (LRET) assay showed that TMP-polyaminocarboxylates bind to Escherichia coli dihydrofolate reductase (eDHFR) fusion proteins with nanomolar affinity in purified solutions and in bacterial lysates. The ability to selectively impart terbium or europium luminescence to fusion proteins in complex physiological mixtures bypasses the need for specific antibodies and simplifies sample preparation. PMID- 21619069 TI - Canolol: a promising chemical agent against oxidative stress. AB - The OOH radical scavenging activity of canolol (CNL) has been studied in aqueous and lipid solutions, using the density functional theory. CNL is predicted to react about 3.6 times faster in aqueous solution than in lipid media. The overall rate coefficients are predicted to be 2.5 * 10(6) and 6.8 * 10(5) M(-1) s(-1), respectively. The OOH radical scavenger activity of canolol is predicted to be similar to that of carotenes, higher than that of allicin, and much higher than that of melatonin. Branching ratios for the different channels of reaction are reported for the first time. It was found that the reactivity of canolol toward OOH radicals takes place almost exclusively by H atom transfer from the phenolic moiety in canolol, regardless of the polarity of the environment. Taking into account that the reactivity of peroxyl radicals is significantly lower than that of other reactive oxygen species, canolol is proposed to be a very good antioxidant. PMID- 21619070 TI - Simple guanidinium salts revisited: room-temperature ferroelectricity in hydrogen bonded supramolecular structures. AB - Dielectric, calorimetric, and X-ray diffraction methods have been employed to characterize the crystals of guanidinium tetrafluoroborate and guanidinium perchlorate, both built of two-dimensional honeycomb hydrogen-bonded sheets. The room-temperature ferroelectricity of these isosymmetric complexes (space group R3m) has been evidenced by the polarization switching in an external electric field and pyroelectric effect. The analysis of structural data as a function of temperature showed that the high values of spontaneous polarization of about 8.5 MUC cm(-2) originate mainly from the ionic displacements, while the exceptional thermally induced increase of polarization is related with the apparent weakening of the N-H...F/N-H...O hydrogen bonds at elevated temperatures. An excellent correlation between the donor-acceptor distance and the relative displacement of the ions in the crystal lattice along the polar direction has been found. The huge entropy change at the two-closely spaced high-temperature phase transitions in guanidinium perchlorate, together with the large crystal polarization, suggest a large electrocaloric effect, the property strongly desired for solid-state cooling applications. PMID- 21619071 TI - Computational model for predicting experimental RNA and DNA nearest-neighbor free energy rankings. AB - Hydrogen-bonding, intrastrand base-stacking, and interstrand base-stacking energies were calculated for RNA and DNA dimers at the MP2(full)/6-311G** level of theory. Standard A-form RNA and B-form DNA geometries from average fiber diffraction data were employed for all base monomer and dimer geometries, and all dimer binding energies were obtained via single-point calculations. The effects of water solvation were considered using the PCM model. The resulting dimer binding energies were used to calculate the 10 unique RNA and 10 unique DNA computational nearest-neighbor energies, and the ranking of these computational nearest neighbor energies are in excellent agreement with the ranking of the experimental nearest-neighbor free energies. These results dispel the notion that average fiber diffraction geometries are insufficient for calculating RNA and DNA stacking energies. PMID- 21619072 TI - Strategies in functional poly(ester amide) syntheses to study human coronary artery smooth muscle cell interactions. AB - The design of new generation cardiovascular biomaterials focuses on biomimetic properties that are capable of eliciting specific cellular responses and directing new tissue formation. Synthetic poly(ester amide)s (PEAs) containing alpha-amino acid residues have the potential to elicit favorable cellular responses. Furthermore, they are biodegradable owing to the incorporation of naturally occurring amino acids. In this study, a family of PEAs was synthesized from selected alpha-amino acids using both solution and interfacial polymerization approaches to optimize their properties for vascular tissue engineering applications. By careful selection of the monomers and the polymerization approach, high-molecular-weight PEAs with low glass-transition temperatures were obtained. Human coronary artery smooth muscle cells (HCASMCs) cultured directly on bare PEA films attached and spread well up to 7 days of culture. Moreover, cell viability was significantly enhanced on all nonfunctional PEAs compared with tissue culture polystyrene controls. The trifluoroacetic acid salt of the lysine-containing functional PEAs was found to retard cell growth but still supported cell viability up to 5 days of culture. Immunostaining of HCASMCs revealed strong vinculin expression, suggesting that the HCASMCs initiated cellular processes for focal adhesion contacts with all PEA surfaces. Conversely, smooth muscle alpha-actin expression was not abundant on the PEA surfaces, suggesting a proliferative smooth muscle cell phenotype. Altogether, our results indicate that these PEAs are promising materials for vascular tissue engineering scaffolds. PMID- 21619073 TI - Metal paste nanocomposite electrodes as a new generation of metallic electrodes. AB - Fabrication of metal paste nanocomposite electrodes is introduced using metal nanostructures and ionic liquids. The combined application of unique properties of nanomaterials and ionic liquids in the design of these metal paste nanocomposites results in electrodes with interesting advantages compared to the conventional metal disk electrodes. In contrast to conventional metallic electrodes, which are usually prone to fouling effects and suffer from weak repeatability and reproducibility, these metal paste nanocomposite electrodes have very exciting advantages. Ease of electrode fabrication; cleaning and activating the electrode surface, together with high electrocatalytic activity; increased degree of active area and surface roughness; antifouling effect; good signal-to-noise ratio; low cost; and low weight are among the advantageous features of these electrodes. Compared to dropping mercury electrodes that have high toxicity, these metal paste nanocomposite electrodes have much less toxicity. Such abilities promote new opportunities for a wide range of electrochemical, sensing, and biosensing applications. PMID- 21619074 TI - Multicompartmental LC-Q-TOF-based metabonomics as an exploratory tool to identify novel pathways affected by polyphenol-rich diets in mice. AB - Metabonomics has recently been used to study the physiological response to a given nutritional intervention, but such studies have usually been restricted to changes in either plasma or urine. In the present study, we demonstrate that the use of LC-Q-TOF-based metabolome analyses (foodstuff, plasma, urine, and caecal content metabolomes) in mice offer higher order information, including intra- and intercompartment relationships. To illustrate this, we performed an intervention study with three different phenolic-rich extracts in mice over 3 weeks. Both unsupervised (PCA) and supervised (PLS-DA) multivariate analyses used for pattern recognition revealed marked effects of diet in each compartment (plasma, urine, and caecal contents). Specifically, dietary intake of phenolic-rich extract affects pathways such as bile acid and taurine metabolism. Q-TOF-based metabonomics demonstrated that the number of correlations is higher in caecal contents and urine than in plasma. Moreover, intercompartment correlations showed that caecal contents-plasma correlations are the most frequent in mice, followed by plasma-urine ones. The number of inter- and intracompartment correlations is significantly affected by diet. These analyses reveal the complexity of interorgan metabolic relationships and their sensitivity to dietary changes. PMID- 21619075 TI - Single nanoskived nanowires for electrochemical applications. AB - In this work, we fabricate gold nanowires with well controlled critical dimensions using a recently demonstrated facile approach termed nanoskiving. Nanowires are fabricated with lengths of several hundreds of micrometers and are easily electrically contacted using overlay electrodes. Following fabrication, nanowire device performance is assessed using both electrical and electrochemical characterization techniques. We observe low electrical resistances with typical linear Ohmic responses from fully packaged nanowire devices. Steady-state cyclic voltammograms in ferrocenemonocarboxylic acid demonstrate scan rate independence up to 1000 mV s(-1). Electrochemical responses are excellently described by classical Butler-Volmer kinetics, displaying a fast, heterogeneous electron transfer kinetics, k(0) = 2.27 +/- 0.02 cm s(-1), alpha = 0.4 +/- 0.01. Direct reduction of hydrogen peroxide is observed at nanowires across the 110 pM to 1 mM concentration range, without the need for chemical modification, demonstrating the potential of these devices for electrochemical applications. PMID- 21619076 TI - Structure-based design of short peptide ligands binding onto the E. coli processivity ring. AB - The multimeric DNA sliding clamps confer high processivity to replicative DNA polymerases and are also binding platforms for various enzymes involved in DNA metabolism. These enzymes interact with the clamp through a small peptide that binds into a hydrophobic pocket which is a potential target for the development of new antibacterial compounds. Starting from a generic heptapeptide, we used a structure-based strategy to improve the design of new peptide ligands. Chemical modifications at specific residues result in a dramatic increase of the interaction as measured by SPR and ITC. The affinity of our best hits was improved by 2 orders of magnitude as compared to the natural ligand, reaching 10( 8) M range. The molecular basis of the interactions was analyzed by solving the co-crystal structures of the most relevant peptides bound to the clamp and reveals how chemical modifications establish new contacts and contributes to an increased affinity of the ligand. PMID- 21619077 TI - Identification and quantification of DNA repair proteins by liquid chromatography/isotope-dilution tandem mass spectrometry using their fully 15N labeled analogues as internal standards. AB - Oxidatively induced DNA damage is implicated in disease, unless it is repaired by DNA repair. Defects in DNA repair capacity may be a risk factor for various disease processes. Thus, DNA repair proteins may be used as early detection and therapeutic biomarkers in cancer and other diseases. For this purpose, the measurement of the expression level of these proteins in vivo will be necessary. We applied liquid chromatography/isotope-dilution tandem mass spectrometry (LC MS/MS) for the identification and quantification of DNA repair proteins human 8 hydroxyguanine-DNA glycosylase (hOGG1) and Escherichia coli formamidopyrimidine DNA glycosylase (Fpg), which are involved in base-excision repair of oxidatively induced DNA damage. We overproduced and purified (15)N-labeled analogues of these proteins to be used as suitable internal standards to ensure the accuracy of quantification. Unlabeled and (15)N-labeled proteins were digested with trypsin and analyzed by LC-MS/MS. Numerous tryptic peptides of both proteins were identified on the basis of their full-scan mass spectra. These peptides matched the theoretical peptide fragments expected from trypsin digestion and provided statistically significant protein scores that would unequivocally identify these proteins. We also recorded the product ion spectra of the tryptic peptides and defined the characteristic product ions. Mixtures of the analyte proteins and their (15)N-labeled analogues were analyzed by selected-reaction monitoring on the basis of product ions. The results obtained suggest that the methodology developed would be highly suitable for the positive identification and accurate quantification of DNA repair proteins in vivo as potential biomarkers for cancer and other diseases. PMID- 21619078 TI - Bioinformatics analysis of a Saccharomyces cerevisiae N-terminal proteome provides evidence of alternative translation initiation and post-translational N terminal acetylation. AB - Initiation of protein translation is a well-studied fundamental process, albeit high-throughput and more comprehensive determination of the exact translation initiation sites (TIS) was only recently made possible following the introduction of positional proteomics techniques that target protein N-termini. Precise translation initiation is of crucial importance, as truncated or extended proteins might fold, function, and locate erroneously. Still, as already shown for some proteins, alternative translation initiation can also serve as a regulatory mechanism. By applying N-terminal COFRADIC (combined fractional diagonal chromatography), we here isolated N-terminal peptides of a Saccharomyces cerevisiae proteome and analyzed both annotated and alternative TIS. We analyzed this N-terminome of S. cerevisiae which resulted in the identification of 650 unique N-terminal peptides corresponding to database annotated TIS. Furthermore, 56 unique N(alpha)-acetylated peptides were identified that suggest alternative TIS (MS/MS-based), while MS-based evidence of N(alpha)-acetylation led to an additional 33 such peptides. To improve the overall sensitivity of the analysis, we also included the 5' UTR (untranslated region) in-frame translations together with the yeast protein sequences in UniProtKB/Swiss-Prot. To ensure the quality of the individual peptide identifications, peptide-to-spectrum matches were only accepted at a 99% probability threshold and were subsequently analyzed in detail by the Peptizer tool to automatically ascertain their compliance with several expert criteria. Furthermore, we have also identified 60 MS/MS-based and 117 MS based N(alpha)-acetylated peptides that point to N(alpha)-acetylation as a post translational modification since these peptides did not start nor were preceded (in their corresponding protein sequence) by a methionine residue. Next, we evaluated consensus sequence features of nucleic acids and amino acids across each of these groups of peptides and evaluated the results in the context of publicly available data. Taken together, we present a list of 706 annotated and alternative TIS for yeast proteins and found that under normal growth conditions alternative TIS might (co)occur in S. cerevisiae in roughly one tenth of all proteins. Furthermore, we found that the nucleic acid and amino acid features proximate to these alternative TIS favor either guanine or adenine nucleotides following the start codon or acidic amino acids following the initiator methionine. Finally, we also observed an unexpected high number of N(alpha) acetylated peptides that could not be related to TIS and therefore suggest events of post-translational N(alpha)-acetylation. PMID- 21619079 TI - In situ measurements of ion-exchange processes in single polymer particles: laser trapping microspectroscopy and confocal fluorescence microspectroscopy. AB - Ion-exchange processes of a cationic dye (Rhodamine B; RhB) were studied for individual polymer particles (diameter of 16-20 MUm) by laser trapping microspectroscopy and confocal fluorescence laser microspectroscopy. The absorbance of RhB at 565 nm adsorbed on a cation-exchange particle increased linearly with the concentration of RhB in the aqueous phase, while it was independent of the particle diameter. Fluorescence intensity profile measurements of RhB along the particle diameter by confocal fluorescence microspectroscopy directly proved that ion exchange took place in the surface layer (~2-MUm thickness) of the particle in the initial stage (1 h). Diffusion of RhB in the particle was very slow, and ion exchange proceeded gradually to the inner volume in the order of days. The ion-exchange processes were analyzed on the basis of simulation of the time course of the concentration profile of RhB in the particle, and the diffusion coefficient of RhB was determined to be (2-4) * 10( 11) cm(2).s(-1). PMID- 21619080 TI - Compositional and structural studies of vinylidene chloride/vinyl chloride copolymers by pyrolysis gas chromatography. AB - A pyrolysis gas chromatography approach has been developed to study the composition and structure of vinylidene chloride/vinyl chloride copolymers. The composition and number average sequence length, which reflects the monomer arrangement in the polymer, were calculated using formulas that incorporate the pure trimer peak intensities and hybrid trimer peak intensities. Because of the reactivity difference between vinyl chloride and vinylidene chloride monomers, the structure of the polymer has been further investigated on the basis of the percentage of grouped monomers (i.e., the number average sequence length for vinyl chloride and vinylidene chloride repeat units). For the vinylidene chloride/vinyl chloride copolymer examined in this study, the composition and number average sequence length elucidated from the pyrolysis gas chromatography study are compared to the product composition specification and/or the composition measured by (1)H-NMR. PMID- 21619081 TI - Measurement of partition coefficients in waterless biphasic liquid systems by countercurrent chromatography. AB - Countercurrent chromatography (CCC) is a chromatographic separation technique that uses a liquid as a stationary phase. Centrifugal forces are used to immobilize the liquid stationary phase when the liquid mobile phase is pushed through it. In CCC, the solutes are separated according to their liquid-liquid partition coefficients. The solutes studied were the alkylbenzene homologues from benzene to hexylbenzene and some polyaromatic hydrocarbons (PAHs) from naphthalene to coronene. Their liquid-liquid partition coefficients were measured in the five waterless biphasic systems formed by heptane, as the apolar liquid phase of the five biphasic systems, and four dipolar aprotic solvents, dimethyl sulfoxide, dimethylformamide, furfural, and N-methylpyrrolidone, and the polar proton-donor solvent methanol. The coefficients were compared to the corresponding capacity factors obtained by classical liquid chromatography on octadecyl-bonded silica. For the five biphasic solvent systems studied, linear relationships were found between the partition coefficients and the sp(3) and sp(2) hybridized carbon atom number for the alkylbenzene and PAH series, respectively. The sp(2) and sp(3) transfer energies were estimated, and their ratio was used to quantify the solvent selectivity toward aromatic extraction. PMID- 21619082 TI - Retention behavior of ionizable isomers in reversed-phase liquid chromatography: a comparative study of porous graphitic carbon and octadecyl bonded silica. AB - The retention behaviors of 36 positional isomers of ionizable substituted benzene compounds have been compared on two different packing materials: porous graphitic carbon (PGC) and octadecyl bonded silica (ODS) using 35% aqueous acetonitrile as the mobile phase. The effect of the mobile phase pH on the solute retention was studied over a range of pH values from pH 2.0 to 7.0. The retention as a function of pH was modeled using equations based on solute ionization. With PGC, the theoretical equations fitted the observed retention data for each class of solute, indicating that the retention mechanism was uniform over the whole pH range. However, with ODS, only the acidic solutes showed agreement with the theoretical model; for the amine-containing compounds, serious deviations from the theory were observed, suggesting that strongly acidic silanols gave added retention at low mobile phase pH. Overall, PGC demonstrated a higher selectivity toward positional isomers than ODS. This was attributed to the greater steric discriminating ability arising from the flat surface of the PGC compared with the more fluid nature of the ODS bonded phase. PMID- 21619083 TI - Theoretical study of high-performance frontal analysis: a chromatographic method for determination of drug-protein interaction. AB - High-performance frontal analysis (HPFA), a chromatographic method to determine unbound drug concentration in drug-protein binding equilibrium, has been considered on the basis of a theoretical plate model, where a rapid equilibrium of drug-protein binding in the mobile phase in the interstices of packing materials and a chromatographic partition equilibrium of the drug were taken into account simultaneously. When a certain excess volume of drug-protein mixed solution is injected directly into a HPFA column packed with a restricted-access type phase that excludes protein but retains drug in the micropores, the drug is eluted as a zonal peak with a plateau region. The elution profile can be well simulated by the mass balance equation derived according to a relatively simple plate theory concept, which confirms that the drug concentration in the plateau range agrees with the unbound drug concentration in the sample solution. The model was applied to the theoretical and systematic investigation of the dependence of the HPFA profile on several chromatographic conditions and the properties of the sample solution, such as injection volume of sample solution, drug and protein concentrations in sample solution, capacity factor of the drug, theoretical plate number, and binding parameters. The smaller capacity factor and the higher column efficiency lead to the larger plateau volume. The lower drug concentration, the higher protein concentration, and the stronger binding constant, which give the lower unbound drug fraction, lead to the larger plateau volume and allow frontal analysis with a smaller sample size. PMID- 21619084 TI - Quantitative Evaluation of SERS-Active Ag Film Nanostructure by Atomic Force Microscopy. AB - The reliability of an image analysis algorithm for atomic force microscopy (AFM) of thin metal films was evaluated by comparison with manual analysis of images and transmission electron micrographs of Ag films deposited on Formvar-coated Cu grids. In order to extract quantitative nanostructural information using the algorithm discussed herein, the optimal fitting parameters were found to be low pass filtering to reject high-frequency noise, a 5 * 5 point grid for identification of particle maxima, and a linear least-squares fit to a hemispheroidal model of particle shape. Metal particle dimensions were defined from the height and radius of the hemispheroid fit. Due to the close spacing of particles in these Ag films, tip geometry causes the greatest error in the height measurements, rather than width measurements. In addition, the effect of scanning parameters such as scan rate and size, applied load, and humidity on particle count and dimensions was examined. Increasing the scan rate reduced the number of resolvable Ag particles, decreased the apparent particle height, and increased the apparent particle radius. Under conditions of low capillary force, a net repulsive force of ~19 nN resulted in subtle tip-induced changes in the Ag surface morphology. The Ag film surface was damaged at a net repulsive force of ~23 nN. At slow scan rates, the moisture layer did not significantly affect the quality of the AFM images obtained over a broad relative humidity range. Finally, the Ag surface structure was found to be very homogeneous over a relatively large area. PMID- 21619085 TI - Determination of paraquat by square-wave voltammetry at a perfluorosulfonated ionomer/clay-modified electrode. AB - A novel Nafion/clay-modified electrode (NCME) was developed for the determination of paraquat by square-wave cathodic stripping voltammetry. The clay that showed the best performance for the fabrication of the NCME is nontronite (SWa-1, ferruginous smectite). The electrochemical behavior of paraquat showed that the cathodic peak at -0.70 V vs Ag/AgCl permits adequate quantification of the analyte. Linear calibration curves are obtained over the 0-80 ppb range, with a detection limit of 0.5 ppb in pH 8 phosphate buffer solution for 4 min preconcentration time. Various factors influencing the determination of paraquat were thoroughly investigated in this study. The practical analytical utility is illustrated by selective measurements of paraquat in real water samples. PMID- 21619086 TI - One-component room temperature vulcanizing-type silicone rubber-based calcium selective electrodes. AB - New calcium-selective membranes for all-solid-state ion sensors are developed with a highly adhesive one-component room temperature vulcanizing-type silicone rubber (RTV-1-type SR) matrix. The membranes are formulated with 21.6 wt % bis(2 ethylhexyl) adipate, 0.3 wt % tetradodecylammonium tetrakis(p-chlorophenyl)borate (ETH 500), 0.1 wt % potassium tetrakis(p-chlorophenyl)borate, and 0.8 wt % calcium-selective neutral carrier ETH 129 or ETH 1001. Plasticizer added to the RTV-1-type SR matrix not only decreases the bulk membrane resistance but also increases the solubility of electroactive components incorporated in the membrane without significantly deteriorating its adhesive strength. It is found that the lipophilic salt ETH 500 remarkably enhances the calcium selectivity of ETH 129 or ETH 1001 ligands in the SR matrix; the selectivity coefficients ([Formula: see text] by separate solution method, where j = Li(+), Na(+), K(+), or Mg(2+)) for the optimized membranes were below 10(-5). Potentiometric characteristics of planar-type Ag electrodes coated with optimized RTV-1-type SR membranes, e.g., response slope 29.0 +/- 0.5 mV/decade, detection limit below 5.0 * 10(-7) M a(Ca)((2+)), and 2-3 mV of potential drift per day, were virtually the same as those of the corresponding poly(vinyl chloride) membrane-based conventional electrodes, but with greatly enhanced sensor-to-sensor reproducibility and lifetime (3-9 weeks). PMID- 21619087 TI - On-line monitoring of powder blend homogeneity by near-infrared spectroscopy. AB - Near-infrared spectroscopy is evaluated as an on-line technique for monitoring the homogeneity of a pharmaceutical blend during the blending process. Blends containing 10% sodium benzoate (model active), which provided an aromatic functionality typical of many pharmaceutical compounds, 39% microcrystalline cellulose (Avicel PH102), 50% lactose, and 1% magnesium stearate were developed to mimic the properties of an actual pharmaceutical blend. A twin-shell V-blender was modified to allow installation of a diffuse reflectance fiber-optic probe at the axis of rotation, and spectra were collected during three experiments using a commercially available near-infrared spectrophotometer. In each experiment, blender control and spectral data collection were controlled by a compilation of software packages. The experiments detected spectral changes which eventually converged to a point of constant variance. Further analysis of the spectral data showed the blend is homogeneous long before a typical blending period is complete. Near-infrared spectroscopy has proven to be a feasible and effective method for the "real time" noninvasive determination of homogeneity in a pharmaceutical blend. PMID- 21619088 TI - Detection and confirmation of N-nitrosodialkylamines using liquid chromatography electrospray ionization coupled on-line with a photolysis reactor. AB - Simultaneous detection and confirmation of several N-nitrosodialkylamines are accomplished by on-line coupling of a photolysis reactor with an HPLC electrospray ionization mass spectrometer. Several parameters such as irradiation wavelength, irradiation time, mobile-phase composition, and pH, as well as different organic acid modifiers are investigated, and their impact on the detection of the N-nitrosodialkylamine-acid complex and its dissociative photolysis products is presented here. Additionally, the type of structural information obtained from the photolytic processes of N-nitrosodialkylamines is compared to that obtained by using in-source collision-induced dissociation. To demonstrate the potential of this technique, six N-nitrosodialkylamines are studied to determine the linearity of the response, the limits of detection and confirmation, and the reproducibility. The technique's versatility is also exhibited by utilizing negative-ion mode as a complementary means for analysis of the compounds. Finally, an illustrative application for N-nitrosodimethylamine analysis in beer is described. PMID- 21619089 TI - Determination of elemental compositions from mass peak profiles of the molecular ion (m) and the m + 1 and m + 2 ions. AB - The relative abundances of M + 1 and M + 2 ions help to identify the elemental composition of the molecular ion (M). But scan speed, sensitivity, and resolution limitations of mass spectrometers have impeded determination of these abundances. Mass peak profiling from selected ion recording data (MPPSIRD) provided faster sampling and enhanced sensitivity, which permitted use of higher resolution. M + 2 profiles having only a few percent of the ion abundance of M were monitored at 20 000 resolution. The relative abundances, exact masses, and shapes of M, M + 1, and M + 2 mass peak profiles were determined. By applying five criteria based on these quantities, elemental compositions were determined even for ions too large (up to 766 Da) to be uniquely assigned from their exact mass and accuracy limits alone. A profile generation model (PGM) was written to predict these resolution dependent quantities by considering all M + 1 and M + 2 ions for each candidate composition. The model also provided assurance that no other compositions were possible. Characterization of the M + 1 and M + 2 profiles by MPPSIRD and the PGM greatly expanded the practical ability of high-resolution mass spectrometry to determine elemental compositions. PMID- 21619090 TI - Determination of dissolved and adsorbed EDTA species in water and sediments by HPLC. AB - This paper describes a method for determining EDTA species in various environmental samples at low molar concentrations by high-performance liquid chromatography (HPLC). Distinction between Fe(III)EDTA and all the other species can be made. NiEDTA can be detected semiquantitatively. The fraction of EDTA adsorbed to suspended particles or to sediments can be determined after desorption with phosphate. After complexation with Fe(III), the EDTA is detected by reversed-phase ion-pair liquid chromatography as the Fe(III)EDTA complex at a wavelength of 258 nm. The behavior of a variety of metal-EDTA complexes during analysis was checked. Determination of different EDTA species (Fe(III)EDTA, NiEDTA, and adsorbed EDTA) is possible in river water, groundwater, and effluents from wastewater treatment plants. Fe(III)EDTA was found to be the main species, at 30-70%; NiEDTA was <10% in most of the samples. Adsorbed EDTA was detected in suspended particles from rivers and wastewater treatment plants and in sediment cores from a lake. The method is suitable for a variety of different samples with different concentration ranges. PMID- 21619091 TI - Precise Measurement of Isotope Ratios with a Double-Focusing Magnetic Sector ICP Mass Spectrometer. AB - The potential of a commercially available double-focusing magnetic sector ICP mass spectrometer (Element, Finnigan MAT, Bremen, Germany) for precise isotope ratio measurement at the low-resolution setting (R = 300) was evaluated. Optimization of scanning conditions led to a relative standard deviation for a set of 10 consecutive 2 min measurements of ~0.1% ((206)Pb(+)/(207)Pb(+)) at signal intensities of ~200 000 counts/s (peak height). This compares favorably with the best values ever reported for quadrupole ICPMS and barely exceeds the theoretical value (counting statistics). Increasing the signal intensity to values >=500 000 counts/s (peak height) resulted in a further reduction of the RSDs obtained (for both (25)Mg(+)/(26)Mg(+) and (206)Pb(+)/(207)Pb(+)) to typically 0.04%. These figures are remarkably better than those reported for commercially available quadrupole ICPMS systems. This improvement significantly reduces the difference between isotope ratio precision of ICPMS on one hand and those of thermal ionization mass spectrometry and plasma source multiple collector mass spectrometry on the other. PMID- 21619092 TI - Pulsed-gas glow discharge for ultrahigh mass resolution measurements with fourier transform ion cyclotron resonance mass spectrometry. AB - A new pulsed-gas glow discharge (GD) source has been developed for use with an external ion source Fourier transform ion cyclotron resonance (FTICR) mass spectrometer. With pulsed argon gas introduction into the GD source, the gas load and pressure in the mass analyzer region were greatly reduced; this resulted in improved mass resolution. Mass resolution of greater than 1 450 000 (fwhm) has been achieved for Cu(+) ions from a brass sample, the highest reported for any type of GD mass spectrometer. The pulsed-gas GD source promises analytical usefulness for ultrahigh resolution measurements in GD mass spectrometry. PMID- 21619093 TI - Analytical chemistry in a drop. Solvent extraction in a microdrop. AB - An organic microdrop (~1.3 MUL) is suspended inside a flowing aqueous drop from which the analyte is extracted. The drop-in-drop system is achieved by a multitube assembly. The aqueous phase is continuously delivered to the outer drop and is aspirated away from the bottom meniscus of the drop. After the sampling/extraction period, a wash solution replaces the sample/reagent in the aqueous layer, resulting in a clear outer aqueous drop housing a colored organic drop containing the extracted material. This also results in an automatic backwash. The color intensity of the organic drop, related to the analyte concentration, is monitored by a light-emitting diode based absorbance detector. After the analytical cycle, the organic drop is removed and replaced by a new one. The performance of the system is illustrated with the determination of sodium dodecyl sulfate (a methylene blue active substance) extracted as an ion pair into chloroform. This unique microextraction system is simple and flexible, permits automated backwashing, consumes only microquantities of organic solvents, and is capable of being coupled with other analytical systems. This concept should prove valuable for preconcentration and matrix isolation in a microscale. PMID- 21619094 TI - Sol-gel-based, planar waveguide sensor for gaseous iodine. AB - A novel sensor for gaseous iodine has been developed using a combination of sol gel processing and planar integrated optical waveguiding technologies. The sensing principle is based on the detection of a charge transfer complex formed between iodine and phenyl groups that have been incorporated into a porous, methylated glass film. The glass film was prepared from siloxane precursors by the sol-gel method. Sensors were fabricated by coating the film over a single mode planar waveguide. Light was coupled into and out of the laminate structure using integral grating couplers, and formation of the charge transfer complex was monitored as attenuated total reflection of the guided wave. The sensor exhibits a linear response to I(2) in the range of 100 ppb to 15 ppm with response and recovery times less than 15 s. The response is selective to 4 ppm iodine in the presence of 10 ppm chlorine and is stable for at least 3 months. PMID- 21619095 TI - Optimization method for simultaneous kinetic analysis. AB - Kinetic analysis is often carried out for simultaneous determinations; thus, a theory to establish its optimal conditions is necessary. A very simple and fast model to find conditions for optimum analytical performance and to predict the quality of simultaneous kinetic analysis has been developed. It is general and applicable to any reaction order or rate constant. The model has been based on the angle between the kinetic vectors and on their norm ratio, which are readily calculated for any kinetic scheme. Evaluation of the proposed model has been carried out by detailed simulations of numerous experimental conditions and analysis by full PCR calculations (when applicable) or nonlinear least-squares fitting. An important conclusion is that analytical performance is determined to a large extent by the stability of the space spanned by the relevant component vectors (in addition to experimental noise and other factors). The quality of the analysis is governed by the angle between the kinetic vectors, while the norm ratio determines the error distribution between components. Optimum conditions for simultaneous kinetic analysis have been studied in several representative examples, regarding the timing of the kinetic monitoring and the effects of concentrations and of rate constant ratios, as well as several other factors of experimental relevance. PMID- 21619096 TI - Assessing the accuracy of analytical methods using linear regression with errors in both axes. AB - In this paper, a new technique for assessing the accuracy of analytical methods using linear regression is reported. The results of newly developed analytical methods are regressed against the results obtained using reference methods. The new test is based on the joint confidence interval for the slope and the intercept of the regression line, which is calculated taking the uncertainties in both axes into account. The slope, intercept, and variances which are associated with the regression coefficients are calculated with bivariate least-squares regression (BLS). The new technique was validated using three simulated and five real data sets. The Monte Carlo method was applied to obtain 100 000 data sets for each of the initial simulated data sets to show the correctness of the new technique. The application of the new technique to five real data sets enables differences to be detected between the results of the joint confidence interval based on the BLS method and the results of the commonly used tests based on ordinary least-squares or weighted least-squares regression. PMID- 21619097 TI - Fabrication and characterization of sputtered-carbon microelectrode arrays. AB - This paper describes a robust and reliable process for fabricating a novel sputter-deposited, thin-film carbon microelectrode array using standard integrated circuit technologies and silicon micromachining. Sputter-deposited carbon films were investigated as potential candidates for microelectrode materials. The surface properties and cross section of the microelectrode arrays were studied by atomic force microscopy and scanning electron microscopy, respectively. Electrical site impedance, crosstalk, and lifetime (dielectric integrity) of microelectrodes in the array were characterized. Electrochemical response of the microelectrodes to hexaammineruthenium(III) chloride and dopamine were investigated by fast-scan cyclic voltammetry and high-speed, computer-based chronoamperometry; results show that thin-film carbon microelectrodes are well behaved electrochemically. The thin carbon films offer extremely good electrical, mechanical, and chemical properties and thus qualify as viable candidates for various electroanalytical applications, particularly acute neurophysiological studies. PMID- 21619098 TI - Ohmic drop compensation in voltammetry: iterative correction of the applied potential. AB - A new method of ohmic potential drop correction for use with potential step and sweep voltammetric methods is described. The method, iterative correction of the applied potential (ICAP), essentially replaces an electronic positive feedback correction with a digital positive feedback correction. This paper will present one form of the ICAP procedure, in which voltammetric current data acquired by a high-speed digital oscilloscope are used to iteratively generate a compensating potential waveform that is then synthesized by an arbitrary waveform generator. The ICAP method is advantageous for high-speed measurements, since many of the difficulties caused by electronic positive feedback compensation are eliminated. PMID- 21619099 TI - Determination of copper in environmental matrices following vapor generation. AB - Copper was determined in environmental matrices following generation, separation, and atomization of a volatile species formed by the merging of an acidified sample solution with an aqueous sodium tetrahydroborate solution at room temperature. The copper species, as yet unidentified, was phase separated in a conventional gas-liquid separator and directed via a stream of Ar carrier gas to an inductively coupled plasma atomic emission detection system. Optimum conditions for generation were investigated. The efficiency of generation/transfer was estimated to be 50%, and no interference from the presence of 1000 mg/L concentration of As, Cd, Co, Ni, Fe, Cr, Mn, Pb, Se, and Zn concomitants was evident. Simple aqueous standards were used for calibration purposes, and good agreement was obtained with certified values in the analysis of National Research Council of Canada marine sediment BCSS-1 and lobster hepatopancreas tissue TORT-1. PMID- 21619100 TI - Macromolecule analysis based on electrophoretic mobility in air: globular proteins. AB - Globular proteins ranging in molecular mass from 5.7 to 669 kDa were separated and analyzed using an aerosol technique based on the electrophoretic mobility of singly-charged molecular ions in air. The ions were produced by electrospraying and drying 100-nm-diameter droplets of a liquid suspension of the proteins, using ionized air to remove the droplet charge due to the spray process. The electrophoretic mobility was measured using a modified commercial continuous-flow differential mobility analyzer operated near atmospheric pressure. An unmodified commercial condensation particle counter was used for detection. The concentrations analyzed ranged from 0.02 to 200 MUg of protein/mL of buffer, with a liquid sample flow rate of approximately 50 nL/min. Sampling time of 3 min was used for each complete distribution measured. The electrophoretic mobilities measured were determined entirely from air flow rates, apparatus geometry, and applied potentials. Results were expressed as electrophoretic mobility equivalent diameters using a Millikan formula. PMID- 21619101 TI - Liquid chromatography combined with tandem mass spectrometry for the confirmation of sarafloxacin in catfish tissue. AB - A specific chromatographic LC/MS/MS assay is described for the confirmatory identification of residues of sarafloxacin, an arylfluoroquinolone antibacterial agent, in catfish tissue. This confirmatory method takes advantage of the specificity provided by sample preparation, liquid chromatography, and tandem mass spectrometry. This kind of multidimensional analysis is commonly used in environmental, pharmacokinetic, residue, and other studies. However, we demonstrate the addition of a previously unreported criterion, the use of ion ratio ranges from the tandem mass spectrometry (MS/MS) experiment as an aid in confirmation. Using the described method, we were able to achieve MS/MS product ion ratios with <7% variation during 1 day of analysis for over 25 injections. We believe the addition of this criterion will increase the scientific certainty of the confirmatory method. PMID- 21619102 TI - Mass spectrometry of proteins directly from polyacrylamide gels. AB - The direct combination of thin-layer gel electrophoresis and matrix-assisted laser desorption/ionization mass spectrometry has been demonstrated with good sensitivity and mass accuracy, offering potential advantages in speed and reduced complexity. Mass spectra have been obtained from isoelectric focusing, sodium dodecyl sulfate, and native gels with as little as 660 fmol of alpha- and beta chain bovine hemoglobin and 1 pmol of horse heart myoglobin loaded. CNBr digests were performed in situ, and the products were probed in-gel. Noncovalent complexes such as multimeric protein systems, enzyme inhibitor complexes, and protein-ligand complexes can also be characterized when gel electrophoresis is run under nondenaturing conditions. This approach shows promise for simplifying the interface between gel electrophoresis and mass spectrometry. PMID- 21619103 TI - Design and Optimization of a Corona Discharge Ion Source for Supercritical Fluid Chromatography Time-of-Flight Mass Spectrometry. AB - The interfacing of capillary column supercritical fluid chromatography (SFC) to time-of-flight mass spectrometry (TOFMS) through atmospheric pressure chemical ionization (APCI) was investigated. An ion source chamber and a new, flexible, and efficient transfer line from the SFC to the TOFMS system were designed to accommodate the requirements of this study. Ionization of analytes was performed using a corona discharge needle. The interface was equipped with two multiple axis translation stages for positioning of the transfer line tip and the discharge needle inside the ion chamber. The investigations were oriented toward the optimization of parameters which have a strong effect on the intensity and stability of the analyte signal, including background stability, corona discharge needle positioning in the ion source, transfer line tip and discharge needle relative positioning, curtain gas and makeup gas flow interactions, ion chamber temperature, and elution pressure of analytes from the SFC system. PMID- 21619104 TI - Enhancement of Separation Efficiency in Capillary Electrophoresis by Electrostacking without Liquid Contact. AB - Quantitative reproducibility and separation efficiency in high-performance capillary electrophoresis is often limited by the reproducibility and the nature of the sample introduction procedure. This is particularly true where electrostacking is involved, whether such is deliberately carried out or happens automatically with the choice of ionic strength of the running electrolyte. Extraneous dispersion frequently originates in the act of sample introduction, for example, from the shock of the capillary entering a sample vial or a sample filled capillary entering the carrier electrolyte source vial. A method based on the stacking of the sample constituents, before the capillary actually contacts the liquid in a vial, can reduce this problem. This technique can be utilized for the preconcentration of sample constituents and to improve separation efficiency by reducing dispersion. In favorable cases, plate counts can increase by as much as 160%, approaching theoretical diffusion-limited efficiencies. PMID- 21619105 TI - Improvement of Gas Chromatographic Performance of Cyclic Siloxane Bonded Phases by Incorporation of n-Alkyl Groups. AB - Cyclic siloxane-based silica columns of high retention capacity for light hydrocarbons were prepared by incorporation of octyl and octadecyl groups on the siloxane skeleton. Spectroscopic studies revealed the presence of the alkyl groups on the silica surface, which results in high surface coverage of the packing materials. The retention characteristics of the columns are significantly higher than those of cyclic siloxane phases due to their high surface coverage and better solute-stationary phase interactions. In agreement with previous data for micropacked columns, the H vs u curves, on passing through the optimum region, remained substantially constant at high linear gas velocities, indicating their usefulness for fast analysis of light hydrocarbons at high flow rates without any appreciable loss in column performance. Presumably due to greater availability of the bonded alkyl groups for interactions, a 10-component mixture of C(1)-C(4) saturated and unsaturated hydrocarbons was sufficiently separated on the hexamethylcyclotrisiloxane-octadecylsilyl phase compared to the conventional octadecyldimethylsilyl phase, irrespective of the high carbon content of the latter. PMID- 21619106 TI - Complexation of Be(II) Ion with 1-(2,4-Dihydroxy-1-phenylazo)-8-hydroxy-3,6 naphthalenedisulfonate as the Chemical Basis of the Selective Detection of Ultratrace Be by Reversed-Phase High-Performance Liquid Chromatography. AB - The complexation reactions of beryllium(II) ion with 1-(2,4-dihydroxy-1 phenylazo)-8-hydroxy-3,6-naphthalenedisulfonate (H-resorcinol) are studied. The acid dissociation constants of H-resorcinol, H(3)L(2-), at 293 K and I = 0.10 [K(OH, NO(3))] are pK(a)((1)) = 5.67, pK(a)((2)) = 8.57, and pK(a)((3)) > 13. The formation constant at 293 K and I = 0.10 [(K,H)NO(3)] is estimated to be log[{[Be(HL)(2-)][H(+)](2)}/{[Be][H(3)L(2-)]}] = -4.58, and pK(a)' = 6.39 for [Be(HL)](2-), which give the basis for the optimization of the precolumn chelation reactions and the masking system with EDTA. The kinetic data for ligand substitution reactions with sulfosalicylate ion are also reported to demonstrate the remarkable inertness of the Be chelate, which is suitable for HPLC separation. Reported is an accurate method for determining traces of Be(II) ion at nanomolar levels with photometric detection coupled with ion-pair reversed phase HPLC. The chelate, [Be(II)L](3-), is efficiently separated on an Asahipak ODP-50 column using tetrabutylammonium bromide as an ion-pairing agent in a methanol (35 wt %)-water eluent. Only Al and Fe give peaks under the conditions used. The large molar absorptivity of the H-resorcinol chelate, 3.99 * 10(4) M( 1) cm(-1) at 500 nm, and the short retention time with excellent peak resolution ensure the ultralow detection limit (3sigma blank) down to 7.2 ppt (0.8 nM) with no preconcentration procedures. The excellent toughness toward the matrix influence was demonstrated using the model solution for an air-dust sample. The HPLC separation, coupled with the EDTA masking procedure, enables one to detect Be(II) ion at 20 nM in the presence of metals at the natural abundance levels in air samples, such as Al, Fe, Ca, Mg, Zn, and Pb at 240, 140, 300, 66, 16, and 6.2 MUM, respectively, in the final solution. PMID- 21619107 TI - Magnetic permeability measurements in bioanalysis and biosensors. AB - A new transducer concept in bioanalysis and biosensors, utilizing measurements of magnetic permeability, is reported. A model system based on dextran ferrofluid, concanavalin A immobilized to a carrier (Sepharose), and glucose was used to demonstrate the feasibility of this approach. Direct ferromagnetic detection of the dextran ferrofluid was achieved by using a measuring coil (transducer) in a Maxwell bridge. A sensitivity of 21 MUV/(MUg Fe/mL) and a rsd value of 3.8% were obtained (n = 5). It was also demonstrated that a small, non-ferromagnetic metabolite (glucose) could be detected using a competitive approach. With an increasing concentration of glucose (20-40 mM), we observed a decrease in the response (0.59-0.11 mV). Reference measurements performed on Sepharose without the biorecognition element, concanavalin A, showed no significant response (0.01 mV). Some potential advantages and drawbacks of this novel type of magnetic transducer are discussed. The advantages include very low interference from the sample matrix, as the transducer is only sensitive to ferromagnetic substances, which rarely are present in samples. In addition, it is suggested that these transducers should be free from fouling. The new transducers are proposed to provide the basis for a new group of affinity biosensors suitable for in vivo and in vitro use. PMID- 21619108 TI - Response to Comment on "Influence of the Chain Length and Surface Density on the Conformation and Mobility of n-Alkyl Ligands Chemically Immobilized onto a Silica Surface". PMID- 21619109 TI - Determination of 2-(4-Morpholinyl)benzothiazole in Environmental Samples by a Gas Chromatograph Equipped with a Flame Photometric Detector. AB - 2-(4-Morpholinyl)benzothiazole (24MoBT) exists in automobile tire rubber as an impurity of a vulcanization accelerator and has been proposed as a potential molecular marker of street runoff (Spies, R. B.; Andresen, B. D.; Rice, D. W., Jr. Nature 1987, 327, 697-699). The present paper describes an analytical method for 24MoBT in environmental samples (e.g., street dusts and river sediments) by gas chromatography. The method relies upon extraction with a 6:4 (v/v) mixture of benzene and methanol, purification by acid extraction, and adsorption column chromatography, followed by determination using capillary GC equipped with a sulfur-selective detector (i.e., FPD). The recovery of 24MoBT for the entire procedure was 85%, and the relative standard deviation for four replicated analyses was 1.5%. The detection limit was 0.08 ng injected 24MoBT, corresponding to 0.20 ng/g of dry sample. The selectivity and sensitivity of the present method permit the determination of 24MoBT at the trace levels (e.g., ~ng/g) encountered in environmental samples. 24MoBT concentrations in various environmental samples are also reported. PMID- 21619110 TI - In situ measurements of ion-exchange processes in single polymer particles: laser trapping microspectroscopy and confocal fluorescence microspectroscopy. PMID- 21619111 TI - Simultaneous Temperature and Holdup Time Ratio Optimization for Tandem Column Tunable Selectivity with High-Speed GC. AB - Analysis time for high-speed, capillary column GC is reduced by the use of a pressure-tunable tandem combination of a nonpolar poly(methylsiloxane) (DB-5) column and a polar trifluoropropyl poly(methylsiloxane) (Rtx-200) column operated isothermally at an optimized oven temperature. By adjusting the pressure at the midpoint between the two columns, the residence times of all sample components are adjusted to give the maximum resolution of the critical component pair. The result is a two-dimensional optimization where the column temperature and the midpoint pressure were adjusted to give the shortest possible analysis time. A previously defined relative resolution function, which requires only empirical capacity factor data for the target compounds, is used as the dependent variable in the optimization algorithm. The resulting three-dimensional resolution map is projected parallel to the relative resolution axis in order to obtain a useful two-dimensional display from which the optimal operating conditions can be determined. PMID- 21619112 TI - Separation orthogonality in temperature-programmed comprehensive two-dimensional gas chromatography. AB - In a comprehensive two-dimensional gas chromatograph, a thermal modulator serially couples two columns containing dissimilar stationary phases. The secondary column generates a series of high-speed secondary chromatograms from the sample stream formed by the chromatogram eluting from the primary column. This series of secondary chromatograms forms a two-dimensional gas chromatogram with peaks dispersed over a retention plane rather than along a line. The method is comprehensive because the entire primary column chromatogram is transmitted through the secondary column with fidelity. One might expect that a two dimensional separation in which both dimensions are basically the same technique, gas chromatography, would be inefficient because the two dimensions would behave similarly, generating peaks whose retentions correlate across dimensions. Applying a temperature program to the two columns, however, can tune the separation to eliminate this inefficiency. The temperature program reduces the retentive power of the secondary column as a function of progress of the primary chromatogram such that the retention mechanism of the primary column is eliminated from the second dimension. Retention of a substance in the second dimension is then determined by the difference in its interaction with the two stationary phases. Retention times in the second dimension then fall within a fixed range, and the whole retention plane is accessible. In a properly tuned comprehensive two-dimensional chromatogram, retention times in the two dimensions are independent of each other, and the two-dimensional chromatogram is orthogonal. Orthogonality is important for two reasons. First, an orthogonal separation efficiently uses the separation space and so has either greater speed or peak capacity than nonorthogonal separations. Second, retention in the two dimensions of an orthogonal chromatogram is determined by two different and independent mechanisms and so provides two independent measures of molecular properties. PMID- 21619113 TI - Sequential method for the field determination of nanomolar concentrations of dimethyl sulfoxide in natural waters. AB - A procedure for the determination of dimethyl sulfoxide (DMSO), along with dimethyl sulfide (DMS) and dimethyl sulfoniopropionate (DMSP), at nanomolar levels in natural waters has been developed. After removal of DMS by purge and cryotrapping, DMSP is removed by the same method after alkaline hydrolysis, and DMSO is reduced to DMS using a combination of sodium borohydride and hydrochloric acid. The DMS produced is stripped, cryotrapped, and analyzed by gas chromatography. Detection of 3 pmol of DMSO was achieved, resulting in a detection limit of 0.05 nM for a 50 mL sample. Mean yield for standards in the range 0.7-130 nM (n = 31) was 95%, and mean precision (as coefficient of variation) was 14%. Precision for replicates of natural seawater samples was always <=10%. Mean yields of the sequential analysis of DMS + DMSP + DMSO (1.5-25 nM) standard mixtures in seawater were >90% for the three species. Filtered seawater samples stored frozen (-20 degrees C) showed no significant changes in DMSO concentration. Since DMSP is the only compound, other than DMSO, that gives rise to DMS upon reduction with NaBH(4), tests were performed to ensure that DMSP is quantitatively removed before the DMSO analysis. Boranes were identified as the substances that produced two major peaks in the chromatogram of the reduction products. Adequate chromatographic conditions were established to avoid their coelution with DMS. This entire protocol allows the sequential determination of DMS, DMSP, and DMSO in natural waters and is suitable for field work, so it should be very useful in studies of dimethyl sulfur biogeochemistry. Some examples of DMSO (along with DMS and DMSP) measurements in the Mediterranean Sea are presented. PMID- 21619114 TI - Decomposition of Terpenes by Ozone during Sampling on Tenax. AB - The ozonolytic decomposition of terpenes and terpenoids during sampling on Tenax is investigated in an artificial air system at ambient concentration levels. The detrimental effect of 8-150 ppbv ozone depends on the chemical structure of the compounds: saturated terpenoids such as 1,8-cineole, camphor, and bornyl acetate are unaffected by ozone. The terpenes and terpenoids which containing one C-C double bond are slightly decomposed in the order camphene < beta-pinene ~ myrtenal < alpha-pinene < sabinene ~ citronellal. The compounds containing two or more double bonds are significantly decomposed in the order d-limonene ~ citral < linalool < beta-ocimene < terpinolene << alpha-terpinene ~ beta-caryophyllene. For alpha-pinene, sabinene and d-limonene, their ozonolysis products are found on the tubes: pinonaldehyde, 5-(1-methylethyl)bicyclo[3.1.0]hexan-2-one, and 3-(1 methylethenyl)-6-oxoheptanal. The analytical recoveries are significantly enhanced for many compounds when the sampling duration is reduced from 10 min to 30 s, explained by the time available for ozonolysis. A miniature ozone scrubber with multiple layers of MnO(2)-coated copper nets was developed and thoroughly tested. The optimal number of plies is found to be 8, which ensures quantitative recoveries for all test compounds except alpha-terpinene, beta-caryophyllene, citral, and citronellal. The results that are reported here call into question previous data on terpenes and terpenoids and/or their oxidation products where measurements have been carried out without the prior removal of ozone. PMID- 21619115 TI - Instrumental requirements for nanoscale liquid chromatography. AB - Nanoscale liquid chromatography (nano-LC), with packed columns of typically 75 MUm i.d. * 15 cm length, packed with C18, 5 MUm of stationary phase, and optimal flow rates of 180 nL/min, can be considered as a miniaturized version of conventional HPLC. Using the down-scaling factor, which corresponds to the ratio of the column diameter in square, (d(conv)/d(micro))(2), excellent agreement between the theoretically calculated values and the values obtained using the down-scaling factor (~3800) has been observed. This factor was applied to all system components, including flow rate, injection and detection volumes, and connecting capillaries. Down-scaling of a conventional HPLC system to a nano-LC system is easy to realize in practice and involves using a microflow processor for nanoflow delivery (50-500 nL/min), a longitudinal nanoflow cell (<=3 nL), a microinjection valve (<= 20 nL), low-dispersion connecting tubing, and zero dead volume connections. Excellent retention time reproducibility was measured with RSD values of +/-0.1% for isocratic and +/-0.2% for gradient elution. Plates counts of more than 100 000/m indicate the excellent performance of the entire nano-LC system. With minimal detectable amounts of proteins in the low femtomole and subfemtomole ranges (e.g., 520 amol for bovine serum albumin), high mass sensitivity was found, making nano-LC attractive for the microcharacterization of valuable and/or minute proteinaceous samples. Coupling nano-LC with concomitant mass spectrometry using nanoscale ion spray or electrospray interfaces looks very promising and is obviously the next step for future work. PMID- 21619116 TI - Ion chromatographic separation of rare-Earth elements using a nitrilotriacetate type chelating resin as the stationary phase. AB - The chromatographic retention behavior of rare-earth elements (REEs) on a chelating resin having a nitrilotriacetate group (NTA gel) was evaluated. The capacity factors for a series of REEs on the NTA gel were in fairly good agreement (R(2) = 0.978) with the stability constants of the corresponding NTA complexes. The NTA gel was applied to the column stationary phase for the ion chromatographic separation of REE. A favorable separation of a series of REEs was achieved within 15 min using a gradient elution of nitric acid. The separated REE ions were detected using the postcolumn derivatization method with chlorophosphonazo III as a colorization reagent. The present chromatographic system, interfaced with inductively coupled plasma mass spectrometry, was applied for the simultaneous determination of 14 REEs. PMID- 21619117 TI - Solid-phase microextraction coupled with high-performance liquid chromatography for the determination of alkylphenol ethoxylate surfactants in water. AB - Nonionic surfactants of the alkylphenol ethoxylate class are relatively nonvolatile analytes that are typically analyzed by LLE and HPLC or GC after derivatization. Solid-phase microextraction (SPME) has seen many GC applications, but recently the ability to hyphenate SPME with HPLC was investigated. In this paper, a new SPME/HPLC method for analysis of Triton X-100 and other alkylphenol ethoxylates is described. Normal-phase gradient elution with detection by UV absorbance at 220 nm was used for the analysis. Several new coated phases, currently in the developmental stage, were evaluated. New Carbowax/template resin and Carbowax/divinylbenzene coatings allowed successful analysis of alkylphenol ethoxylates with a linear range of 100-0.1 mg/L. These coatings provided the best agreement between the ethoxamer distribution of surfactants after extraction and that in the original surfactant. Limits of detection of the individual ethoxamers were determined to be in the low microgram per liter to submicrogram per liter range. Some applications of the method have been demonstrated for sewage sludge samples. PMID- 21619118 TI - Determination of Dansyl Amino Acids and Oxalate by HPLC with Electrogenerated Chemiluminescence Detection Using Tris(2,2'-bipyridyl)ruthenium(II) in the Mobile Phase. AB - A new electrogenerated chemiluminescence detection method is investigated for use in detection in reversed-phase and reversed-phase ion-pair HPLC with Ru(bpy)(3)(2+) in the mobile phase. In this method, different concentrations of Ru(bpy)(3)(2+) are dissolved in the mobile phase and the HPLC column flushed with the mobile phase for 1 h until the column is saturated with Ru(bpy)(3)(2+). The separated analytes along with Ru(bpy)(3)(2+) pass through an optical electrochemical flow cell which has a dual platinum electrode held at a potential of 1250 mV vs a Ag/AgCl reference electrode. On the surface of the electrode, Ru(bpy)(3)(2+) is oxidized to Ru(bpy)(3)(3+) which reacts with the analytes to emit light. The retention times, retention orders, detection limits, and linearity in working curves are compared to those obtained with the conventional postcolumn Ru(bpy)(3)(2+) addition method. The retention times for dansyl amino acids with Ru(bpy)(3)(2+) in the mobile phase are longer than those obtained with the postcolumn addition approach. This may be caused by pi-to-pi interactions between the aromatic groups of the dansyl derivatives and the bipyridyl groups of Ru(bpy)(3)(2+) in the Ru(bpy)(3)(2+)-saturated reversed-phase column. Similarly, oxalate is separated from urine and blood plasma samples by reversed-phase ion pair HPLC. Plasma samples are obtained using ultrafiltration to remove proteins from whole blood. Retention times for oxalate with the two detection techniques are identical, and detection limits for these techniques are compared. PMID- 21619119 TI - Influence of charge regulation in electrostatic interaction chromatography of proteins. AB - Recently, the "slab model" was proposed to describe the interaction between a protein and the charged stationary phase surface in electrostatic interaction chromatography. The model is based on the solution of the linearized Poisson Boltzmann equation for a system consisting of two charged planar surfaces in contact with an electrolyte solution. In the model it is assumed that the charge densities of both the protein and the stationary phase are constant during the adsorption process. However, as the protein comes close to the oppositely charged stationary phase surface, the protein net charge will change due to the electrical field from the stationary phase. In this paper, the theory for charge regulation is applied to the original slab model, and simple algebraic equations are developed in order to include the effect of charge regulation on the capacity factor. A large body of retention data are reanalyzed with the new model, and it is found that there is good agreement between the chromatographically and titrimetrically obtained protein net charge. An interesting consequence of charge regulation is that it gives a contribution to the retention of proteins with zero net charge and even to proteins with the same sign of charge as the stationary phase. PMID- 21619120 TI - Determination of mean diameter and particle size distribution of acrylate latex using flow field-flow fractionation, photon correlation spectroscopy, and electron microscopy. AB - Flow field-flow fractionation (flow FFF) was employed to determine the mean diameter and the size distribution of acrylate latex materials having diameters ranging from 0.05 to 1 MUm. Mean diameters of the samples determined by flow FFF are in good agreement with those obtained from photon correlation spectroscopy (PCS). Scanning electron microscopy (SEM) yielded a mean diameter that is about 20% lower than those obtained from flow FFF or PCS, probably due to the shrinkage of particles during sample drying and high-vacuum measurements. It was found that flow FFF is particularly useful for the determination of particle size distributions of latex materials having broad size distributions. Flow FFF separates particles according to their sizes and yields an elution curve that directly represents the particle size distribution of the sample. In PCS, measurements had to be repeated at more than one scattering angle to obtain an accurate mean diameter for the latex having a broad size distribution. Flow FFF was fast (less than 12 min of run time) and showed an excellent repeatability in measuring the mean diameter with +/-5% relative error. PMID- 21619121 TI - Determination of sulfur dioxide in wine using a quartz crystal microbalance. AB - A new method for the analysis of both total and bound SO(2) in wine is proposed, based on a quartz crystal microbalance (QCM), and it is compared with the widely used Ripper method. The proposed method is faster than the Ripper's, and the instrumentation is either home-made or widely available. When both methods are applied to the same sample, the results obtained using the QCM method are bracketed in an interval less than one-tenth the size of that obtained using the Ripper method. Although the SO(2) concentrations found using the QCM method correlate well with the ones obtained with the Ripper method, the results are systematically higher, which can be explained as due to the absence of interferences known to affect the Ripper method. PMID- 21619122 TI - Selection of the best calibration sample subset for multivariate regression. AB - This paper discusses a methodology for selecting the minimum number of calibration samples in principal component regression (PCR) analysis. The method uses only the instrumental responses of a large set of samples to select the optimal subset, which is then submitted to chemical analysis and calibration. The subset is selected to provide a low variance of the regression coefficients. The methodology has been applied to UV-visible spectroscopy data to determine Ca(2+) in water and near-IR spectroscopy data to determine moisture in corn. In both cases, the regression models developed with a reduced number of samples provided accurate results. As far as precision is concerned, a similar root-mean-squared error of cross-validation (RMSECV) is found when comparing the new methodology with the results of the regression models that use the complete set of calibration samples and PCR. The number of analyzed samples in the calibration set can be reduced by up to 50%, which represents a considerable reduction in costs. PMID- 21619123 TI - Selectivity and Related Measures for nth-Order Data. AB - Analytical figures of merit are often used as criteria to decide whether or not a given instrumental method is suitable for attacking an analytical problem. To date, figures of merit primarily exist for analytical instruments producing data indexed by one variable, i.e., first-order instruments and first-order data. Almost none exist for instruments that generate data indexed by two variables, i.e., second-order instruments and data, and none exist for instruments supplying data indexed by three or more variables, i.e., nth-order instruments and data. This paper develops practical mathematical tools that can be used to create several figures of merit for nth-order instrumentation, namely, selectivity, net analyte signal, and sensitivity. In particular, the paper fully develops a local selectivity measure for second-order instrumentation and tests its performance using simulated second-order data and real second-order data obtained by gas chromatography with Fourier transform infrared detection and liquid chromatography with photodiode array detection. Also included in the paper is a brief discussion on practical uses of nth-order figures of merit. PMID- 21619124 TI - Implementation of Electrochemically Synthesized Silver Nanocrystallites for the Preferential SERS Enhancement of Defect Modes on Thermally Etched Graphite Surfaces. AB - Highly oriented pyrolytic graphite (HOPG) surfaces, on which atomically well defined roughness has been introduced via high-temperature gasification reactions, are investigated by noncontact mode atomic force microscopy (NC-AFM) and Raman spectroscopy both before and after the electrochemical deposition of silver nanocrystallites on these surfaces. Exposure of freshly cleaved HOPG surfaces to an O(2)-rich ambient at 650 degrees C for a few minutes caused the formation of 1-monolayer-deep, circular etch pits on the HOPG basal plane surface. Silver nanocrystallites were electrochemically deposited onto these etched surfaces at two coverages: 0.5 mC cm(-)(2) (or 5 nmol of Ag(0) cm(-)(2)) and 2.4 mC cm(-)(2) (25 nmol of Ag(0) cm(-)(2)). At the lower coverage, NC-AFM images revealed that silver decorated only the circumference of the circular etch pits, forming a uniform annular ring with an apparent diameter of 200-250 A and a height of ~15 A. At the higher silver coverage, an increase in the height but not the diameter of this annulus was observed, and additional silver nanostructures [Formula: see text] having dimensions of 300-350 A diameter and 15 A height [Formula: see text] were observed on atomically smooth regions of the graphite basal plane. The Raman spectroscopy of these surfaces was investigated and compared with spectra for nanocrystallite-modified but unetched HOPG basal plane surfaces and thermally etched surfaces on which no silver was deposited. For for thermally etched HOPG surfaces at either silver coverage, SERS-augmented Raman spectra were obtained in which defect modes of the graphite surface [Formula: see text] derived from "finite" graphite domains at the surface [Formula: see text] were strongly and preferentially enhanced. In addition, an enhanced band near 2900 cm(-)(1) was assigned to nu(OH) from carboxylate moieties present at step edges based on the basis of the observed pH dependence of the enhancement. PMID- 21619125 TI - Complexation between Copper(II) and Glycine in Aqueous Acid Solutions by Window Factor Analysis of Visible Spectra. AB - Window factor analysis (WFA), a self-modeling chemometric technique, is used to extract the concentration profiles of the complexes formed between Cu(II) and glycine (GlyH) during pH titration in the acid region. The visible absorption spectra of aqueous acid solutions, each containing 0.0020 M Cu(II) and 0.500 M glycine, are recorded at various pHs, ranging from 1 to 7. The concentration profiles and the spectral absorptivities of four copper species, Cu(2+), CuGlyH(2+), CuGly(+), and CuGly(2), are obtained. The spectral absorptivities of the complexes are in good agreement with those reported in the literature. The concentration profiles are used to determine the stepwise equilibrium constants of the three complexes. The reason why rank annihilation evolving factor analysis (RAEFA), a precursor technique, was not successful in resolving the spectral data is explained. PMID- 21619126 TI - Development of a room-temperature phosphorescence fiber-optic sensor. AB - The design of a new fiber-optic sensor based on solid-surface room-temperature phosphorimetry is presented for the analyses of polycyclic aromatic hydrocarbons in water samples. Analytical figures of merit are given for several compounds of environmental importance. Limits of detection at the nanograms per milliliter level were estimated for pyrene, benzo[e]pyrene, benzo[ghi]perylene, 1,2:3,4 dibenzanthracene, coronene, and 2,3-benzofluorene. The linearity of response of the phosphorescence sensor was evaluated, showing a fairly linear behavior for quantitative analysis. Finally, the feasibility of monitoring polycyclic aromatic hydrocarbons in aqueous media was illustrated by identifying pyrene in a contaminated groundwater sample. PMID- 21619127 TI - Rate-limiting steps of tyrosinase-modified electrodes for the detection of catechol. AB - The response currents obtained for tyrosinase-modified Teflon/graphite, carbon paste, and solid graphite electrodes in the presence of catechol are analyzed primarily using rotating disk electrode experiments. The rate-limiting steps, such as the electrochemical reduction of o-quinones and the enzymatic reduction of oxygen as well as the enzymatic oxidation of catechol, are theoretically considered and experimentally demonstrated for the different electrode configurations. PMID- 21619128 TI - H(2)O(2)-generating peroxidase electrodes as reagentless cyanide sensors. AB - Inexpensive, reagentless, and simple (single-electrode) cyanide biosensors are developed using a pyrolytic graphite (PG) electrode on which horseradish peroxidase (HRP) is adsorbed. The electrode is poised at -300 mV vs Ag/AgCl for 40 s to reduce dissolved O(2) to H(2)O(2) at the PG surface. The generated H(2)O(2) accumulates in the diffusion layer. The potential is then stepped to 0 mV, at which the accumulated H(2)O(2) is reduced, though the O(2) reduction does not proceed. Since the H(2)O(2) reduction is catalyzed by HRP, the transient cathodic current is inhibited by cyanide. Therefore, the transient current is a function of the cyanide concentration. A HRP/PG electrode with saturated HRP coverage is reliable, and it can determine 10(-)(5)-10(-)(3) M cyanide. On the other hand, the electrode with lower HRP coverage is less reliable, though it is so sensitive as to detect 2 * 10(-)(7) M cyanide because the system is under kinetic control. PMID- 21619129 TI - Apparent Formation of an Oxidant by Electrochemical Reduction in the Mercury(0,I,II) Chloride System. AB - The paradoxical appearance of a cathodic reaction sometimes observed in anodic stripping voltammetry and stripping potentiometry when using mercury film electrodes in chloride media containing mercury(II) has been investigated by systematically varying relevant chemical and electrochemical parameters and comparing the results with thermodynamic equilibrium calculations. Microscopic observations of morphological changes on the electrode surface caused by potential variations were made possible by using a novel electrode design. Three conditions have to be fulfilled for the cathodic reaction to occur: (a) formation of calomel by reaction between elemental mercury on the electrode surface and mercury(II) in solution, (b) subsequent reduction of mercury(II) to elemental mercury on the calomelized electrode surface, and (c) a chloride concentration in the range 0.001-3.5 M. Different ways of avoiding the interference from the cathodic reaction in stripping voltammetry and stripping potentiometry are experimentally demonstrated, and a mechanism for the appearance of the cathodic reaction is proposed. PMID- 21619130 TI - A Glucose Biosensor Based on Immobilization of Glucose Oxidase in Electropolymerized o-Aminophenol Film on Platinized Glassy Carbon Electrode. AB - A high-performance amperometric glucose biosensor has been developed, based on immobilization of glucose oxidase in an electrochemically synthesized, nonconducting poly(o-aminophenol) film on a platinized glassy carbon electrode. The large microscopic surface area and porous morphology of the platinized glassy carbon electrode result in high enzyme loading, and the enzyme entrapped in the electrodeposited platinum microparticle matrix is stabler than that on a platinum disk electrode surface. The response current of the sensor is 20-fold higher than that of the sensor prepared with a platinum disk electrode of the same geometric area. The experiments showed that the high sensitivity of the sensor is due not only to the large microscopic area but also to the high efficiency of transformation of H(2)O(2) generated by enzymatic reaction to current signal on the platinized glassy carbon electrode. The response time of the sensor is <4 s, and its lifetime is >10 months. PMID- 21619131 TI - Dynamics and performance of fast linear scan anodic stripping voltammetry of cd, pb, and cu using in situ-generated ultrathin mercury films. AB - The dynamics of fast linear scan (LS) ASV for the simultaneous detection of Cd, Pb, and Cu was investigated at various scan rates (0.5-10 V/s) and at different metal ion concentrations (50-800 nM) utilizing ultrathin mercury films (9 nm) at a conventional size (d(0) = 1 mm) electrode. Results of the investigation show that when the thin films were utilized, diffusion of metals through the mercury film was not the rate-limiting step of the stripping process at moderate to fast scan rates (0.5-10 V/s). A fairly linear relationship between the peak height and scan rate was observed at scan rates (0.5-10 V/s) beyond the upper limit of the theoretical model for the behavior of LS-ASV. In addition, peak width at half height (b(1/2)) as low as 33 mV was achieved at 0.5 V/s. The behavior of LS-ASV in terms of peak width at these scan rates is thus different from what the theoretical model of LS-ASV would have predicted. For the ultrathin mercury films, at least two additional factors, kinetics and concentration, have significant effects on practical LS-ASV. Experimental results show that the stripping process of Cu was primarily kinetic-controlled for fast scans, while those for Cd and Pb were dependent on both scan rates and concentrations. The ultrathin mercury film resulted in a significant enhancement of the ratio of signal-to-baseline slope (i(p)/Deltai(b), a ratio used to measure the effectiveness of discrimination of the peak signal against the steep sloping baseline in LS-ASV) for Cd and Pb stripping peaks, but only a slight enhancement for Cu stripping peaks. The optimal performance of LS-ASV in terms of sensitivity, peak width, and enhancement of the i(p)/Deltai(b) ratio for the three metals was achieved at 2 V/s. Because of the high reproducibility of the background currents of the stable in situ MTFs, background subtraction was carried out at 2 V/s with little hysteresis. This feature, combined with the enhancement of the i(p)/Deltai(b) ratio at the fast scan rate of 2 V/s, allowed for the detection of sub-ppb levels of Cd, Pb, and Cu at a deposition time of 2 min. PMID- 21619132 TI - Temperature-jump investigation of alkyl chain length effects on sorption/desorption kinetics at reversed-phase chromatographic interfaces. AB - The influence of the alkyl chain length on the kinetics of solute retention at reversed-phase chromatographic surfaces is examined. A Joule-discharge temperature-jump relaxation experiment was used to monitor reversible sorption/desorption kinetics at C4- and C8-modified silica/solution interfaces. Biexponential sorption/desorption relaxation kinetics were observed for a charged fluorescent probe, 1-anilino-8-naphthalenesulfonate (ANS), on both C4- and C8 silica surfaces. Both relaxation rates on C4 surfaces were sufficiently slow to be measured and increased linearly with solute concentration. One of the relaxations on a C8 surface is too fast to be resolved from the heating rate, similar to behavior of the solute on a longer chain C18-silica. These observations suggest that sorption kinetics on the intermediate chain length surfaace, C4-silica, are different from kinetics on longer chain length surfaces, C8- and C18-silica. From a fit of the data to a two-step kinetic model, the rates of both adsorption and partition of the ionic probe on the C4 chain are estimated; both rates exhibit significant influence over the equilibrium constant. The relaxation rate of a neutral probe, N-phenyl-1-naphthylamine, is also measured; the results indicate a fast (diffusion-controlled) adsorption step, followed by a detectable barrier to partition that is similar to the partition barrier for ANS on the C4-silica surface. These results show that the alkyl chain length of modified silica strongly influences retention kinetics. PMID- 21619133 TI - Rudimentary capillary-electrode alignment for capillary electrophoresis with electrochemical detection. AB - A capillary-electrode holder was constructed for electrochemical detection in capillary electrophoresis (CE). The device allows for positioning of the working electrode at the end of the capillary column without the aid of micropositioners or microscopes. The design facilitates the exchange of electrodes and capillaries without the need of refabricating the entire capillary-electrode setup. The system can be assembled in a very short period of time. Alignment with the self guided system proved to be reproducible for the electrodes used (carbon, nickel, copper). The advantages of reduced downtime and low cost, make the device very attractive for the routine analysis of electroactive species by CE with electrochemical detection. PMID- 21619134 TI - Flow immunoassay using solid-phase entrapment. AB - A flow injection immunoassay was performed using a column packed with reversed phase sorbents to effect separation of the immunoreacted species by entrapping free analyte and allowing antibody-conjugated analyte to pass unretained. Fluorescein-labeled analyte was measured in a competitive assay for the anticonvulsant drug phenytoin. The simplicity of the assay was the greatest advantage of the technique, which allowed for measurement of phenytoin in a 2-min assay time. The reliable detection limit for the assay was 5 nmol L(-)(1) of phenytoin in serum. The columns were regenerated with periodic injections of ethanol solutions to remove the entrapped analyte and prepare the column for subsequent analyses. PMID- 21619135 TI - Biomagnetic neurosensors. 4. Design and optimization for analytical use. AB - Biomagnetic neurosensors based on magnetic stimulation and magnetic detection of neural events depend critically upon the effective matching of the magnetic transducers and the neural tissue employed. Although the properties of wire wrapped ferrite core transducers can be predicted from electromagnetic fundamentals, meaningful analytical measurements using real nerves as molecular recognition elements require additional calibration and optimization steps in order to achieve good system response and lifetimes. This note provides some design guidelines and experimental test procedures to enable potential users to employ biomagnetic neurosensors in other laboratories. PMID- 21619136 TI - Chemiluminescent low-light imaging of biospecific reactions on macro- and microsamples using a videocamera-based luminograph. AB - The analytical performance of a low-light imaging luminograph for quantitative luminescence analysis was evaluated in terms of sensitivity, spatial resolution, accuracy, precision, and sample geometry, at the macrolevel and in combination with optical microscopy. The system allows for the detection of 400 amol of enzymes such as alkaline phosphatase and horseradish peroxidase using 1,2 dioxetanes and luminol/p-iodophenol or acridancarboxylate esters, respectively, as chemiluminescent substrates. Enzymatic activity and spatial distribution of nylon net immobilized-alkaline phosphatase was studied; the system permits the quantification of the immobilized enzyme with a spatial resolution as low as 1 MUm. Other applications, such as the alkaline phosphatase localization in 8 MUm intestinal mucosa cryosections, quantitative immunocytochemistry, and dot blot DNA hybridization reactions, were studied and optimized. The system was also employed for in situ hybridization assay of cytomegalovirus DNA in infected human fibroblasts. The presence of a viral genome was revealed with digoxigenin-labeled probes and alkaline phosphatase-labeled anti-digoxigenin antibody, using chemiluminescent substrate for this enzyme. The luminescent signal was intense and stable, and the probe was imaged and quantified within single cells with higher intensity in the nuclei, with a spatial resolution as low as 1 MUm and very low background. The results show that this technique is an ultrasensitive and potent analytical tool to localize and quantify biomolecules at microscopic level, and it is suitable for many bioanalytical applications. PMID- 21619137 TI - Grating light reflection spectroscopy for determination of bulk refractive index and absorbance. AB - An optical sensing technique is described and evaluated for sensitivity to changes in refractive index and absorbance of model sample matrices. A binary dielectric/metal transmission diffraction grating is placed in contact with a sample and utilized in reflection mode; thus, the light captured and analyzed does not pass through the sample. This particular condition creates thresholds at which a particular transmitted diffraction order is transformed from a traveling wave to an evanescent one. The positions of these thresholds depend upon the complex dielectric function of the sample, the period of the grating, and the wavelength and incident angle of light striking the grating. Experimental evidence directly supports the theoretical predictions regarding responses to both the real and imaginary portions of the refractive index: the reflection coefficient derivative wavelength peak position shifts linearly with changes in the real part of the refractive index, and the derivative peak amplitudes exhibit a square-root dependence on absorbance. Refractive index sensitivity to a series of ethanol/water solutions is demonstrated with detectable changes in index as small as 2 * 10(-)(6). Absorbance sensitivity is shown via the differentiation of methylene blue samples having equivalent 1 cm path length absorbances between 0.459 and 244 AU. In a single reflection measurement, GLRS offers a large dynamic range for absorbance detection, allows simultaneous determination of bulk refractive index in optically dense media, and provides a platform for performing continuous process analysis. PMID- 21619138 TI - Holographic sensor for water in solvents. AB - The diffraction color of a gelatin holographic diffraction grating changed as a function of the water activity when immersed in a "wet" hydrophobic liquid. Quantification of the absorption maximum of the diffracted light showed that it was related, after calibration, to either the water content or the water activity of the solvent. The holographic diffraction grating measured water contents of hydrocarbon solvents at sensitivities comparable to that of the Karl Fischer coulometric titrator and over a wide range of water contents. A grating immersed in xylene revealed a visible color change when the water content was increased from 47 to 120 ppm. Conversely, the holographic grating responded to ethanol in water in the range 0-1% (w/w). The inexpensiveness and simplicity of silver halide holographic reflection gratings, combined with their relatively high sensitivity, suggests that these devices might find widespread application as immersible water activity sensors for hydrophobic liquids. PMID- 21619139 TI - Laser-induced breakdown spectrometry of titanium dioxide antireflection coatings in photovoltaic cells. AB - Silicon photovoltaic cells have design and material requirements different from those of most other silicon electronic devices. Not only are nearly ideal silicon surfaces required, but also the bulk properties must be of uniform high quality for high-energy conversion efficiency. In this paper, emission spectra of laser generated plasmas from titanium dioxide antireflection coatings in solar cells are reported. A pulsed nitrogen laser at 337.1 nm was used with a pulse width of 10 ns and a laser fluence of 8.6 J/cm(2) onto the sample. The plasmas were detected using a charge-coupled device. Depth profilings from several samples with different thicknesses of titanium dioxide have been studied. A method for measuring thin TiO(2) films based on the observation of such profiles was developed. The effect of the laser fluence on the sensitivity of the present method has been examined. Depth resolutions are fluence-dependent but are on the order of 40 nm. The dependence between titanium dioxide plasma intensity and material reflectivity at the laser wavelength is discussed. Another valuable aspect of the technique is the ability to perform measurements in a contactless manner at room temperature and atmospheric pressure on wafer-sized samples. PMID- 21619140 TI - Time-resolved inductively coupled plasma mass spectrometry measurements with individual, monodisperse drop sample introduction. AB - Individual ion clouds, each produced in the ICP from a single drop of sample, were monitored using time-resolved mass spectrometry and optical emission spectrometry simultaneously. The widths of the ion clouds in the plasma as a function of distance from the point of initial desolvated particle vaporization in the ICP were estimated. The Li(+) cloud width (full width at halfmaximum) varied from 85 to 272 MUs at 3 and 10 mm from the apparent vaporization point, respectively. The Sr(+) cloud width varied from 97 to 142 MUs at 5 and 10 mm from the apparent vaporization point, respectively. The delays between optical and mass spectrometry signals were used to measure gas velocities in the ICP. The velocity data could then be used to convert ion cloud peak widths in time to cloud sizes in the ICP. Li(+) clouds varied from 2.1 to 6.6 mm (full width at half-maximum) and Sr(+) clouds varied from 2.4 to 3.5 mm at the locations specified above. Diffusion coefficients were estimated from experimental data to be 88, 44, and 24 cm(2)/s for Li(+), Mg(+), and Sr(+), respectively. The flight time of ions from the sampling orifice of the mass spectrometer to the detector were mass dependent and varied from 13 to 21 MUs for Mg(+) to 93 to 115 MUs for Pb(+). PMID- 21619141 TI - Capillary HPLC-NMR Coupling: High-Resolution (1)H NMR Spectroscopy in the Nanoliter Scale. AB - Coupling HPLC and NMR is one of the most powerful techniques for simultaneous separation and structural elucidation of unknown compounds in mixtures. To date, however, minimizing the detection volume, as is required when coupling NMR with miniaturized separation techniques, has been accompanied by a dramatic loss in resolution of the NMR spectra. Here, we report on the coupling of gradient capillary HPLC with on-column, high-resolution NMR detection. On-line stopped flow and static (1)H NMR spectra were acquired with capillary columns of 75-315 MUm i.d. With detection over a length of 1.2 cm, cell volumes cover a range of 50 900 nL. An on-line-detected NMR separation of dansylated amino acids was carried out in a 315 MUm i.d. fused silica capillary packed to a length of 12 cm with C(18) stationary phase. The low solvent consumption makes the use of fully deuterated solvents economically feasible. NMR spectra with resolution on the order of 3 Hz were obtained using a 50 nL detection cell to measure 1.1 nmol of dansylated gamma-aminobutyric acid under static conditions in a 75 MUm i.d. capillary. PMID- 21619142 TI - Quantitative in situ monitoring of an elevated temperature reaction using a water cooled mid-infrared fiber-optic probe. AB - A novel water-cooled mid-infrared fiber-optic probe is described which is heatable to 230 degrees C. The probe has chalcogenide fibers and a ZnSe internal reflection element and is compact and fully flexible, allowing access to a wide range of standard laboratory reaction vessels and fume cupboard arrangements. Performance is demonstrated via the in situ analysis of an acid-catalyzed esterification reaction in toluene at 110 degrees C, and the results are compared with those from a conventional extractive sampling loop flow cell arrangement. Particular emphasis is given to the quantitative interpretation of the spectroscopic data, using gas chromatographic reference data. Calibration data are presented for univariate and partial least squares models, with an emphasis on procedures for improving the quality of interpreparation calibration and prediction through the use of focused reference analysis regimes. Subset univariate procedures are presented that yield relative errors of <5%, and bias corrected partial least squares procedures are described that result in relative errors of interpreparation calibration and prediction consistently <3%. This paper demonstrates the considerable power of fiber-optic mid-IR spectroscopy combined with bias correction partial least squares procedures for the efficient in situ quantitative analysis of laboratory scale reactions. PMID- 21619143 TI - Glow discharge mass spectrometric analysis of atmospheric particulate matter. AB - A direct current (dc) glow discharge mass spectrometer has been used to analyze atmospheric particulate matter. The sample preparation used is simple and time saving. The air is sucked by a pump through a single-orifice impactor stage, in which the aerosols are impacted on a metal support, forming a central spot. This metal plate is directly used as a cathode in a dc glow discharge mass spectrometer. Evaluation of the sample loading and of the discharge parameters allowed us to optimize the signal intensity and to minimize its decrease, the latter being a consequence of its consumption by continuous sputtering in the discharge. The available aerosol analysis time could be prolonged to more than 3 h, a time span necessary to perform a multielement analysis using a magnetic sector instrument and long integration times. A NIST reference aerosol was measured to evaluate the quantitative analysis potential. The internal reproducibility was better than 10% RSD, and the limits of detection were estimated to be in the low ppm or sub ppm region. Even without the use of any standards or correction factors, glow discharge mass spectrometry could offer good semiquantitative results, based only on the use of an internal standard. PMID- 21619144 TI - Aerosol MALDI with a Reflectron Time-of-Flight Mass Spectrometer. AB - Aerosol matrix-assisted laser desorption/ionization (MALDI) has been combined with a reflectron time-of-flight mass spectrometer for improved mass resolution. A methanol solution of matrix and analyte was sprayed directly into a reflectron time-of-flight mass spectrometer, and the aerosol particles were irradiated and ionized with a frequency-tripled Nd:YAG laser at 355 nm. Mass resolution of over 300 was observed for the peptides bradykinin, angiotensin II, and gramicidin D and for vitamin B(12). This represents a resolution enhancement of approximately 10-fold over that previously reported for aerosol MALDI with a linear time-of flight instrument. PMID- 21619145 TI - Microsecond pulsed glow discharge as an analytical spectroscopic source. AB - A pulsed glow discharge, operating in the microsecond regime, has been found to be advantageous for the examination of solid samples. We have studied the spectroscopic response by atomic emission, absorption, fluorescence, and mass spectrometries. Results to date show enhanced efficiency for analytical response of the sputtered sample atoms. This type of discharge also permits acquisition of useful diagnostic information concerning glow discharge processes. PMID- 21619146 TI - Analysis of Photoablation Products Resulting from Polymer Materials by Supersonic Beam/Multiphoton Ionization/Time-of-Flight Mass Spectrometry. AB - Photoablation products arising from polymer materials were examined by supersonic beam spectrometry, and the results are compared with those obtained by thermal decomposition. The high selectivity provided by supersonic beam spectrometry allows detection of minor species, e.g., styrene occurred from poly(alpha methylstyrene) by cleavage of a methyl group and by proton rearrangement. Because ablation techniques involve high temperatures, thermally stable materials such as poly(p-methylstyrene) can be examined. The latter material is difficult to examine by thermal decomposition, even at 350 degrees C. It is also possible to differentiate between isomer ablation products, e.g., alpha-methylstyrene and p methylstyrene. The instrumental setup described herein was used to examine several authentic samples, such as ABS resin (acrylonitrile-butadiene-styrene) and O-ring (SBR, styrene-butadiene rubber), as well as polystyrene foam. As a result, a styrene segment could be confirmed to be present in these materials. However, no evidence was found for its presence in a glue sample that was suspected to contain styrene on the basis of conventional mass spectrometry. The latter finding confirms the high degree of selectivity of the technique. PMID- 21619147 TI - Design of chromatographic and electrophoretic separation based on ion-pair formation of aromatic disulfonates with polyammonium ions. AB - Ion-pair formation of polyammonium ions with aromatic disulfonates (ADSs) has been investigated with ion-exchange chromatography. Data obtained imply that polyethyleneammonium ions [NH(3)(+)(CH(2)CH(2)NH(2)(+))(n)H] recognize the lengths between sulfonate groups in ADSs in the absence of specific ion-pair formation. The molecular length of the diethylenetriammonium ion is almost equal to that between sulfonate groups in ADSs tested when this polyammonium ion adopts an all-trans conformation; the match of lengths dominates ion-pair formation. However, anthraquinone-1,8-disulfonate (1,8-AnDS) shows a different behavior. This compound specifically interacts with polyammonium ions having an ethylenediammonium structure in a molecular terminal and forms much more stable ion-pairs than any other ADS. It is speculated that the accommodation of a terminal ammonium ion in the pocket consisting of two sulfonate groups in the 1,8 AnDS molecule allows stable complex formation. These are applied to the enhancement of the selectivity in chromatographic and electrophoretic separation of ADS. PMID- 21619148 TI - Quantitative injection from a microloop. Reproducible volumetric sample introduction in capillary zone electrophoresis. AB - A wire loop deployed at the tip of a capillary electrophoresis system has been investigated as a means of quantitative injection. A thin film of a liquid is formed on the loop, the loop is transferred to a sealed chamber, and then pneumatic pressure is applied to introduce the contents of the loop into the capillary. As long as the applied pressure is below a certain threshold, no air is introduced into the capillary, even after the loop contents have been fully introduced. Sample surface tension and viscosity do not have a significant effect on the injected volume. The small loop injection technique appears to be a robust and reproducible alternative to presently practiced approaches to sample injection in CE. PMID- 21619149 TI - On-line coupling of flow field-flow fractionation and multiangle laser light scattering for the characterization of macromolecules in aqueous solution as illustrated by sulfonated polystyrene samples. AB - Seven sulfonated polystyrene standards (18 000-3 000 000 g/mol), taken as model substances for macromolecular polyelectrolytes, were dissolved in aqueous 0.1 M sodium nitrate solution and characterized by multiangle laser light scattering coupled on-line to flow field-flow fractionation. The distributions of molar mass and root mean square radius and the diffusion coefficients were obtained for each sample using a constant field of force for separation. Relationships between molar mass and root mean square radius [(z)(0.5) = (2.71 * 10( )(2))M(w)(0.56)] or diffusion coefficient [D = (7.10 * 10(-)(8))M(w)(-)(0.68)] were calculated. To investigate the static analytical range of this novel hyphenated technique a mixture of all seven samples was fractionated applying a programmed field. The relationship obtained between root mean square radius and molar mass was used to calculate a Mark-Houwink equation [[eta]calcd = (2.99 * 10(-)(2))M(w)(0.68)]. To verify this result, the intrinsic viscosities for all samples were measured at low shear rate and found to be in good agreement [[eta]calcd = (2.77 * 10(-)(2))M(w)(0.67)]. PMID- 21619150 TI - Fast, accurate mobility determination method for capillary electrophoresis. AB - A new method for accurately determining effective mobilities and electroosmotic flow rates for capillary electrophoresis is described. The proposed method can be performed using most commercial capillary electrophoresis instruments. Problems inherent to the conventional mobility determination method such as a variable electroosmotic flow during the run and migration through unthermostated regions of the capillary are eliminated with the use of the proposed method. In addition, very low effective mobilities and electroosmotic flow rates can be measured quickly and reproducibly. Also, cation mobilities and anion mobilities can be measured in a single run regardless of the magnitude or direction of the electroosmotic flow. PMID- 21619151 TI - Extraction of Volatile PAHs from Air by Use of Solid Cyclodextrin. AB - An approach for extraction of polycyclic aromatic hydrocarbons (PAHs) from air using solid cyclodextrin is presented. A comparison study in which beta cyclodextrin is replaced by alpha-cyclodextrin provides evidence that beta cyclodextrin extracts vapor phase PAHs by formation of inclusion complexes rather than by association or adsorption interactions. Thus, solid cyclodextrin complexes with vapor phase PAHs and thereby reduces their volatilities. The gas solid interaction of the PAHs with beta-cyclodextrin and the effect of beta cyclodextrin on the volatilities of these compounds are discussed. Fluorescence and absorbance spectroscopies are used to examine the variables that affect the formation of the PAH complexes with the solid cyclodextrin. The use of this system for improved ambient air sampling is proposed. PMID- 21619152 TI - Direct resolution of optically active isomers on chiral packings containing ergoline skeletons. 5. Enantioseparation of amino Acid derivatives. AB - A new procedure for ergot alkaloid-based chiral stationary phase preparation is described. Synthesis is based on bonding the allyl derivative of terguride to mercaptopropylsilanized silica gel. The packing exhibits higher content of chiral selector, stability, reproducibility, and enantioselectivity toward amino acids compared to that previously studied. The chromatographic behavior of amino acids with different side chains and substituent groups is investigated in order to obtain a deeper insight into the enantiodiscriminative mechanism as well as to determine the limitations and strengths of terguride as a chiral selector for this class of compounds. A variety of factors, including mobile phase parameters such as pH, ionic strength, content and nature of organic modifier, and temperature, are examined. PMID- 21619153 TI - Liquid chromatographic determination of nitro-substituted polynuclear aromatic hydrocarbons by sequential electrochemical and fluorescence detection. AB - Nitro-substituted polynuclear aromatic hydrocarbons occur in ambient suspended particulate matter and are of special concern because they act as direct mutagens. Determination of these compounds in a complex matrix such as particulate matter present in diesel engine exhaust is complicated [Formula: see text] even mass spectrometry requires initial cleanup and separation steps. We propose a sensitive liquid chromatographic method with an unique selectivity: an electrochemical detector operating in the reductive mode is followed by a fluorescence detector. When the NO(2) group is reduced to the NH(2) group, there is a major increase in fluorescence; nitro-PAH compounds are essentially nonfluorescent. A difference fluorescence signal or a difference chromatofluorogram is generated by subtracting the chromatogram obtained with the electrochemical detector off from that obtained with the electrochemical detector on. Applications to diesel engine exhaust samples are demonstrated. PMID- 21619154 TI - Characterization of the pulsed discharge electron capture detector. AB - A new version of the pulsed discharge electron capture detector (PDECD) has been developed and characterized. Changes to the old version include a slightly altered detector geometry, replacement of the polymer insulation with sapphire and quartz, and the use of methane dopant gas instead of nitrogen or hydrogen. Various operating parameters have been investigated and optimized, including discharge current, dopant gas, bias voltage, and sample introduction position. The resulting detector is more inert and more sensitive (a limit of 36 fg for lindane) and capable of operation at temperatures as high as 400 degrees C. By running 23 halocarbon compounds on the improved PDECD and on a (63)Ni-ECD using the same GC system, we find that the PDECD is superior to (63)Ni-ECD in terms of sensitivity, linearity, and response time. We attribute the enhanced sensitivity to a lower positive ion concentration, which in turn lowers the electron-positive ion rate of recombination. Pesticides (including some real-world samples) have also been analyzed on the PDECD. The results demonstrate that the PDECD can replace the radioactive ECD typically used in these analyses. PMID- 21619155 TI - Embossable grating couplers for planar waveguide optical sensors. AB - Planar optical waveguides are an attractive tool for use in analytical chemistry and spectroscopy. Although similar to fiber optics, planar waveguides have been slow to be commercially accepted due to the difficulty of coupling light into the guide. Generally, prism coupling is the method of choice in the laboratory, as efficiencies approaching 80% can be reached. However, prisms are impractical for routine use for several reasons: expensive positioning equipment is required, coupled power is sensitive to environmental fluctuations, and prism coupling prohibits the fabrication of a truly planar device. The use of thin gratings on the surface of the waveguide allows for a two-dimensional structure to be maintained, while providing enough efficiency to be useful as a sensor. Our research efforts focus on developing a technique to make inexpensive, reproducible gratings that are easy to fabricate. By chemically modifying the surface of a commercial grating with a suitable release agent, it is possible to emboss replica gratings onto a variety of waveguide types. The fabrication of embossed gratings will be described, and their performance on glass, ion diffused, polymer, and semiconductor waveguides will be presented. PMID- 21619156 TI - Two-phase stopped-flow measurement of the protonation of tetraphenylporphyrin at the liquid-liquid interface. AB - The formation rate of the protonated form of tetraphenylporphyrin (TPP) in a dispersed two-phase system composed of dodecane and aqueous trichloroacetic acid (TCA) was studied by means of a stopped-flow method. The protonation reaction took place at the liquid-liquid interface, and the diprotonated TPP (H(2)TPP(2+)) formed was adsorbed there. In order to determine the rate-determining process, changes in absorbance at the absorption maximum wavelengths of TPP and H(2)TPP(2+) were analyzed. The obtained rate constant for the decrease of TPP in the organic phase, 21 +/- 2 s(-1), was in agreement with that for the increase of diprotonated TPP at the interface, 20 +/- 3 s(-1). The observed rate constants did not show any dependence on concentrations of both TPP and the acid. The experimental results suggested the rate-determining step to be the molecular diffusion process of TPP in the stagnant layer in the organic phase side at the liquid-liquid interface, and the thickness of the stagnant layer was estimated as 1.4 * 10(-4) cm. PMID- 21619157 TI - Chemiluminescence of luminol catalyzed by electrochemically oxidized ferrocenes. AB - Ferrocenes oxidized at an indium tin oxide-coated glass electrode catalyze the chemiluminescent reaction of luminol with hydrogen peroxide. The catalytic reaction has been studied with ferrocene derivatives in solution and covalently attached to ovalbumin adsorbed on the electrode. It is shown that chemiluminescence is initiated by electrochemical oxidation of the ferrocene derivative. PMID- 21619158 TI - Preconcentration and Voltammetric Determination of Mercury(II) at a Chemically Modified Glassy Carbon Electrode. AB - In this work, the organic compound 2-mercaptobenzimidazole was covalently bound on the surface of a glassy carbon rod, via silanization, yielding a material capable of selectively complexing Hg(2+) ions. This material was applied as an electrode for voltammetric determination of mercury(II) following its nonelectrolytic preconcentration. After exchanging the medium, the voltammetric measurements were carried out by anodic stripping in the differential pulse mode (pulse amplitude, 50 mV; scan rate, 1.25 mV s(-)(1)) using 10(-)(2) mol L(-)(1) NaSCN solution as supporting electrolyte. An anodic stripping peak was obtained at 0.06 V (vs SCE) by scanning the potential from -0.3 to +0.3 V. After a 5 min preconcentration period in a pH 4.0 Hg(2+) solution, this electrode shows increasing voltammetric response in the range 0.1-2.2 MUg mL(-)(1), with a relative standard deviation of 5% and a practical detection limit of 0.1 MUg mL( )(1) (5.0 * 10(-)(7) mol dm(-)(3)). Compared with the conventional stripping approach, this chemically modified glassy carbon electrode procedure presented good discrimination against interference from Cu(II) in up to 10-fold molar excess. PMID- 21619159 TI - Determination of monomethylcadmium in the environment by differential pulse anodic stripping voltammetry. AB - A differential pulse anodic stripping voltammetric (DPASV) method was used to differentiate between the cadmium species Cd(2+) and MeCd(+) (Me = methyl) in aquatic systems. These two species show peaks in the DPASV voltammogram which differ by 112 mV. In model experiments, it was demonstrated that monomethylcadmium is not stable at pH 2, but under higher pH conditions, normally found in fresh and ocean water samples, the identity of MeCd(+) was verified by different investigations, including cyclic voltammetry, selective extraction of a complex of diethyldithiocarbamate with MeCd(+) into n-hexane, and photochemical dissociation of MeCd(+) by UV irradiation. It was also shown that humic acids do not influence the voltammetric determination of monomethylcadmium. For the first time, it was possible to analyze MeCd(+) in environmental samples. During different expeditions with the German research vessel Polarstern, monomethylcadmium could be determined above the detection limit of 470 pg L(-1) in nearly all surface water samples of the South Atlantic with spot concentrations of up to about 700 pg L(-1), whereas in the North Atlantic only 15 30% of the total samples showed MeCd(+) concentrations above this limit. The existence of MeCd(+) in the remote area of the South Atlantic, as well as positive correlations with the local bioactivity in the ocean, indicates biomethylation as the most probable formation process for this methylated cadmium species. This assumption is supported by the simultaneous occurrence of other methylated heavy metal compounds, such as Me(3)Pb(+). Up to 48% of the total cadmium was found to be monomethylcadmium in some Arctic meltwater ponds. PMID- 21619160 TI - Voltammetric determination of the synthetic pyrethroid insecticide tetramethrin in acetonitrile. AB - The synthetic pyrethroid insecticide tetramethrin may be reduced reversibly (E degrees ' = -1.650 V vs Ag/Ag(+)) in acetonitrile at hanging mercury drop electrodes (HMDE) and glassy carbon electrodes. On the voltammetric time scale, the initial electron-transfer process involves the reversible formation of a radical anion. Data obtained from electron paramagnetic resonance spectroscopy indicate that the unpaired electron of the radical is located within the phthalimide system of the molecule. The radical anion may be further reduced at very negative applied potentials with the number of processes being dependent on the nature of the voltammetric technique. The detection limit (3sigma) for the determination of tetramethrin in acetonitrile at a glassy carbon electrode, using differential pulse voltammetry, was found to be 2.1 * 10(-6) M. At a HMDE the detection limit is lower, having a value of 9.6 * 10(-7) M. The limit of determination (10sigma) at a glassy carbon electrode is 3.5 * 10(-6) M and at a HMDE is 3.0 * 10(-6) M. Tetramethrin was selectively determined in an insecticide formulation, at a glassy carbon electrode using differential pulse voltammetry, at a concentration (w/v) of 0.34 +/- 0.02%. The determined concentration is in good agreement with the stated value of 0.350 +/- 0.018% (w/v). PMID- 21619161 TI - PVC-Based 2,2,2-Cryptand Sensor for Zinc Ions. AB - A PVC-based membrane of 2,2,2-cryptand exhibits a very good response for Zn(2+) in a wide concentration range (from 2.06 ppm to 6.54 * 10(3) ppm) with a slope of 22.0 mV/decade of Zn(2+) concentration. The response time of the sensor is <10 s, and the membrane can be used for more than 3 months without any observed divergence in potentials. The proposed sensor exhibits very good selectivity for Zn(2+) over other cations and can be used in a wide pH range (2.8-7.0). It has also been possible to use this assembly as an indicator electrode in potentiometric titrations involving zinc ions. PMID- 21619162 TI - Detection of microspotted carcinoembryonic antigen on a glass substrate by scanning electrochemical microscopy. AB - Microspots of carbinoembryonic antigen (CEA) on glass substrates were characterized by scanning electrochemical microscopy (SECM). CEA was immobilized via a sandiwch method using horseradish peroxidase (HRP)-labeled anti-CEA. The reduction current of the oxidized form of ferrocenylmethanol generated by the HRP reaction was monitored to view SECM images. This method detects as low as ~10(4) CEA molecules in a single 20-MUm-radius spot. PMID- 21619163 TI - Development of a Solid Phase Carbon Trap for Simultaneous Determination of PCDDs, PCDFs, PCBs, and Pesticides in Environmental Samples Using SFE-LC. AB - The purpose of this study was to find materials that can be used as a solid phase trap in a commercial supercritical fluid extraction instrument. PX-21 active carbon mixed with ODS proved to yield the best results. After SFE of the target compounds, the planar fraction containing the environmental pollutants PCDDs, PCDFs, and planar PCBs was successfully separated on a PX-21 solid phase trap from the nonplanar fraction containing the other PCBs and pesticides. Direct injection of the concentrated fractions on GC/MS was possible without further cleanup. SFE followed by on-line carbon column chromatography was tested using standard solutions (applied on filter paper) as well as biological samples (human adipose tissue). PMID- 21619164 TI - Microscopic laser desorption/postionization fourier transform mass spectrometry. AB - A microscopic laser desorption/postionization Fourier transform mass spectrometer (LD/FTMS) is described. The lateral resolution can be <1 MUm with the inherent FTMS high mass resolution intact. Laser postionization allows a certain selectivity and an increase in sensitivity. This capability should allow materials characterization in a wide variety of cases. We demonstrate microscopic desorption and postionization for atoms and molecules. PMID- 21619165 TI - Integrated microdevice for DNA restriction fragment analysis. AB - An integrated monolithic device (8 mm * 10 mm) that performs an automated biochemical procedure is demonstrated. The device mixes a DNA sample with a restriction enzyme in a 0.7-nL reaction chamber and after a digestion period injects the fragments onto a 67-mm-long capillary electrophoresis channel for sizing. Materials are precisely manipulated under computer control within the channel structure using electrokinetic transport. Digestion of the plasmid pBR322 by the enzyme HinfI and fragment analysis are completed in 5 min using 30 amol of DNA and 2.8 * 10(-3) unit of enzyme per run. PMID- 21619166 TI - Detection of Total Trans Fatty Acids Content in Margarine: An Intercomparison Study of GLC, GLC + TLC, FT-IR, and Optothermal Window (Open Photoacoustic Cell). AB - Four techniques, i.e., gas-liquid chromatography, gas-liquid chromatography + thin-layer chromatography, and two spectroscopic methods, Fourier transform infrared spectroscopy and optothermal window, a variant of the open photoacoustic cell, were intercompared to determine their potential to detect the total trans fatty acid content in margarine. At the same time, this study represents a first application of the optothermal window technique at long wavelengths (10 MUm). The total trans fatty acid data obtained by different methods show good mutual agreement. Besides offering several attractive advantages above conventional methods, the optothermal window also proved suitable for measuring total trans fatty acid content as low as 2%. PMID- 21619167 TI - Ion Chromatography-Photodiode Array UV-Visible Detection of Cr(III) Hydrolytic Polymerization Products in Pure and Natural Waters. AB - Hydrolytic polymerization of Cr(III) aqua ions was investigated in pure Milli-Q water and in surface waters, using Sephadex column fractionation followed by nonsuppressed ion chromatography (IC) with UV detection at lambda 462 nm. Pure and natural waters spiked with 0.04 M Cr(III) were fractionated on Sephadex columns into four fractions using eluents with increasing ionic strength. Fractions were analyzed for total Cr by atomic absorption, and recoveries ranged from 94 to 101%. Fractions representing monomeric and low oligomeric Cr (III) species were subjected to IC using a low-capacity mixed resin column and a mobile phase consisting of 2 M NaClO(4)/0.02 M HClO(4) at pH 4.50. Monomeric Cr(III) species were detected in the IC chromatograms of the freshly prepared Cr(III) solution with capacity factors (k') ranging from 0.05 to 0.40. In the 3-days-aged samples, dimeric and trimeric peaks with k' = 1.09 and 1.70, respectively, were detected. Monomeric, dimeric and trimeric Cr(III) peaks collected from the IC preparative experiments were scanned between lambda 200 and 600 nm, using a photodiode array detector. The UV-visible spectral characteristics of the monomer, dimer, and trimer confirmed their identities. The trimeric Cr(III) peak was more predominant in the IC chromatograms of the surface water and accounted for 6.5-35.9% of the total Cr(III) in the samples. PMID- 21619168 TI - Coupling of reversed-phase liquid column chromatography and fourier transform infrared spectrometry using postcolumn on-line extraction and solvent elimination. AB - An on-line postcolumn extraction module was used in conjunction with a solvent elimination interface for the semi-on-line coupling of reversed-phase liquid chromatography (RP-LC) and Fourier transform infrared spectrometry (FT-IR). The extraction module consisted of a phase segmentor, an extraction coil, and a phase separator. Dichloromethane was used as extraction solvent. The organic phase delivered by the separator was evaporated by a spray-jet assembly that simultaneously deposited the extracted analytes onto a zinc selenide window, which was subsequently analyzed by FT-IR microscopy. The method is evaluated by studying parameters such as postcolumn band broadening, phase separation efficiency, evaporation efficiency, extraction yield, eluent composition, and use of nonvolatile buffer salts. Good-quality spectra were obtained for test compounds (phenylureas and quinones), which were separated by RP-LC using a buffered eluent with high water content. Large-volume injections allowed FT-IR detection at the submicrogram per milliliter level. PMID- 21619169 TI - Lifetime-based sensing: influence of the microenvironment. AB - The influence of the microenvironment on the fluorescence behavior of indicator molecules is investigated. A model is developed to describe the fluorescence decay of indicator molecules in a nonuniform medium. Its consequences for fluorescence lifetime-based chemical sensors are discussed and verified in two examples, namely, a pH sensor using a pyrene compound in a hydrogel and a ruthenium complex for oxygen sensing embedded in a polystyrene membrane. PMID- 21619170 TI - Ion-Pair Extraction of Metalloporphyrins into Acetonitrile for Determination of Copper(II). AB - Equilibrium study of ion-pair extraction of a cationic water-soluble porphyrin [5,10,15,20-tetrakis(1-methylpyridinium-4-yl)porphyrin, H(2)tmpyp(4+)] and its metalloporphyrins (MP) into the acetonitrile layer, separated by addition of sodium chloride (4.00 mol dm(-)(3)) to a 1:1 (v/v) acetonitrile-water mixed solvent, was carried out to develop a new and useful method for the determination of a subnanogram amount of copper(II). M denotes Zn(2+), Cu(2+), Co(3+), Fe(3+), and Mn(3+), and P(2)(-) is porphyrinate ion. The extraction and dissociation constants of the ion-pair complexes, defined by K(ex) = [MP(ClO(4))(4)](org)[MP(4+)](aq)(-)(1)[ClO(4)(-)](aq)(-)(4), K(dis,1) = [MP(ClO(4))(3)(+)](org)[ClO(4)(-)](org)[MP(ClO(4))(4)](org)(-)(1), and K(dis,2) = [MP(ClO(4))(2)(2+)](org)[ClO(4)(-)](org)[MP(ClO(4))(3)(+)](org)(-)(1), were determined by taking into account the partition constant of sodium perchlorate (K(D) = 1.82 +/- 0.01). The equilibrium constants were found to be K(ex)K(dis,1) = (7.2 +/- 1.3) * 10(4), (6.4 +/- 0.9) * 10(4), (1.35 +/- 0.13) * 10(5), (4.8 +/- 0.6) * 10(3), (1.23 +/- 0.05) * 10(4), and (1.42 +/- 0.07) * 10(3) at 25 degrees C for the free base porphyrin (H(2)tmpyp(4+)) and the metalloporphyrins of zinc(II), copper(II), cobalt(III), iron(III), and manganese(III), respectively. The K(dis,2) values were (2.9 +/- 1.4) * 10(-)(2), (3.1 +/- 1.1) * 10(-)(2), (8.0 +/- 4.9) * 10(-)(3), and (5.1 +/- 2.2) * 10(-)(2) for the free base porphyrins and the metalloporphyrins of zinc(II), copper(II), and cobalt(III), respectively. The results were developed for determination of a trace amount of copper(II) (3 * 10(-)(8)-4 * 10(-)(6) mol dm(-)(3)) in natural water samples using H(2)tmpyp(4+) with a molar absorptivity of 3.1 * 10(5) mol(-)(1) dm(3) cm(-)(1) at a precision of 1.3% (RSD). The determination of copper(II) was not interfered by the presence of 10(-)(4) mol dm(-)(3) of Mn(2+), Co(2+), Ni(2+), Hg(2+), Cd(2+), Ag(+), Cr(3+), V(5+), Al(3+), Mg(2+), Ca(2+), Br(-), I(-), SCN(-), and S(2)O(3)(2)(-) and 10(-)(5) mol dm(-)(3) of Fe(3+), Zn(2+), and Pd(2+). PMID- 21619171 TI - A new fuzzy regression algorithm. AB - A new fuzzy regression algorithm is described and compared with conventional ordinary and weighted least-squares and robust regression methods. The application of these different methods to relevant data sets proves that the performance of the procedure described in this paper exceeds that of the ordinary least-squares method and equals and often exceeds that of weighted or robust methods, including the two fuzzy methods proposed previously (Otto, M.; Bandemer, H., Chemom. Intell. Lab. Syst. 1986, 1, 71. Hu, Y.; Smeyers-Verbeke, J.; Massart, D. L. Chemom. Intell. Lab. Syst. 1990, 8, 143). Moreover, we emphasize the effectiveness and the generality of the two new criteria proposed in this paper for diagnosing the linearity of calibration lines in analytical chemistry. PMID- 21619172 TI - A biosensor array based on polyaniline. AB - This paper describes the fabrication of polyaniline-based microsensors and microsensor arrays for the estimation of glucose, urea, and triglycerides. Microelectronics technology has been used to produce gold interdigitated microelectrodes on oxidized silicon wafers. Polymer deposition and enzyme immobilization has been done electrochemically. Electrochemical potential control has been used to direct enzyme immobilization to the chosen microelectrodes and prevent it at other microelectrodes in contact with the enzyme solution. This has enabled the immobilization of three different enzymes on three closely spaced microelectrodes, resulting in a sensor array which can analyze a sample containing a mixture of glucose, urea, and triolein in a single measurement using a few microliters of the sample. This strategy is quite general and can be extended to other enzyme-substrate systems to eventually produce an "electronic tongue". PMID- 21619173 TI - Determination of kinetics of the karl Fischer reaction based on coulometry and true potentiometry. AB - A new measurement technique based on a combination of coulometry and zero-current potentiometry is described for determination of the kinetics of rapidly reacting Karl Fischer (KF) reagents. This makes it possible to determine the order as well as the rate constant for large variations in the concentrations of iodine and water present during a titration. It was shown that for imidazole-based methanolic reagents exposed to a large variation in the concentration of water, the KF reaction is first order with respect to iodine, sulfur dioxide, and water only for reagents in which the concentration of nonprotonated imidazole is very low. The rate constant determined for such a reagent (1 M imidazole, 0.8 M sulfur dioxide, 0.1 M iodine) was equal to that reported earlier in the literature. Regions showing first-order kinetics were also found for low concentrations of water when imidazole concentrations up to 2 mol/L were used, provided that these reagents had a quotient [Im](free)/[ImH(+)] around 4. In the interval 2-8 mol/L of imidazole, the order of the reaction with respect to iodine was, in most cases, one-half, while it was changed to between one-half and one with respect to water. The rate of the KF reaction was found to increase by nearly 5 orders of magnitude for a reagent in which the concentration of nonprotonated imidazole was increased from 0 (rate constant equal to 2.6 * 10(3) L(2) mol(-)(2) s(-)(1)) to about 7 mol/L. For most of these reagents, a recovery rate close to 100% was attained. A high concentration of nonprotonated imidazole in combination with a high concentration of sulfur dioxide could, however, lead to a change in stoichiometry of the KF reaction when larger amounts of water were determined (250 MUg of water added to 3.4 mL of reagent solution). A reaction scheme is proposed which might explain this change in stoichiometry observed for some reagent compositions. By use of the described most rapidly reacting reagents, it was shown to be possible to carry out titrations even at such a low end-point concentration as 10(-)(10) M of iodine within 1-2 min. PMID- 21619174 TI - Characterization of chiral host-guest complexation in fast atom bombardment mass spectrometry. AB - A new method has been developed for the characterization of complexion between host and guest molecules. Adduct formation between chiral crown ethers 1 and 2 and enantiomeric ammonium ions 4 and 5 was examined. The reference compound 3 (achiral host) was chosen to be similar in structure to the chiral crown ethers for quantitative measurements. Our approach is based on a formalism assuming an equilibrium: [chiral host + H](+) + [achiral host + chiral guest](+) ? [chiral host + chiral guest](+) + [achiral host + H](+). The equlibrium constant for this process was calculated using the relative peak intensities of the corresponding species in the FAB mass spectra. It was found that these provide significantly better reproducibility and more reliable results than the relative peak intensity method described before (Sawada, M.; et al. J. Am. Chem. Soc. 1992, 114, 4405; 1993, 115, 7381; Org. Mass Spectrom. 1993, 28, 1525).(1)(-)(3) In the examples studied, the equilibrium constants corresponding to the formation of heterochiral adducts (S,S-R or R,R-S) were higher than those for the formation of homochiral aggregates (S,S-S or R,R-R). PMID- 21619175 TI - Electroanalytical detection of glucose using a cyanometalate-modified electrode: requirements for the oxidation of buried redox sites in glucose oxidase. AB - Modification of a nickel electrode with a mixture of iron and ruthenium cyanometalates allows one to efficiently turn over glucose oxidase in the presence of its substrate. A limit of detection for glucose of 25 MUM can be obtained with an observed saturation concentration of 10 mM. Glucose detection is found to be very sensitive to the cationic environment of the electrolyte. The largest currents for glucose oxidation are observed in the presence of nonelectroactive multiply charged cations. The solid state nature of the surface confined cyanometalate redox mediator argues against the widely held mechanism for enzyme oxidation in which the redox mediator is required to enter the enzyme active site. PMID- 21619176 TI - A nitrite sensor based on a highly sensitive nitrite reductase mediator-coupled amperometric detection. AB - Highly sensitive nitrite sensors have been developed for the first time based on mediator-modified electrodes. Tetraheme cytochrome c nitrite reductase from Sulfurospirillum deleyianum and cytochrome cd(1) nitrite reductase from Paracoccus denitrificans are able to accept electrons from artificial electron donors, which simultaneously act as electron mediators between the enzyme and an amperometric electrode. In addition to methyl viologen, redox-active compounds such as phenazines (phenosafranin, safranin T, N-methylphenazinium, 1-methoxy-N methylphenazinium) and triarylmethane redox dyes (bromphenol blue and red) were selected from a range of redox compounds exhibiting the most efficient performance for nitrite detection. After precipitation, the electron mediators were incorporated in a graphite electrode material. Enzyme immobilization is performed by entrapment in a poly(carbamoyl sulfonate) (PCS) hydrogel. Diffusion coefficients and apparent heterogeneous rate constants of the mediators as well as homogeneous rate constants of nitrite sensors were determined by chronoamperometry and cyclic voltammetry. The phenosafranin-modified electrode layered with the PCS hydrogel immobilization of tetraheme cytochrome c nitrite reductase yielded linear current responses up to 250 MUM nitrite with a sensitivity of 446.5 mA M(-)(1) cm(-)(2). The detection limit of the enzymatic nitrite sensor was found to be 1 MUM nitrite. PMID- 21619177 TI - Conductive polymer films as ultrasensitive chemical sensors for hydrazine and monomethylhydrazine vapor. AB - Thin films of the electrically conductive polymer poly(3-hexylthiophene) were investigated as ultrasensitive chemical sensors for hydrazine and monomethylhydrazine vapor. The threshold limit value for these highly toxic species, which are used extensively as rocket fuels, has recently been lowered to 10 ppb for 8-h exposure, necessitating the development of instrumentation with improved sensitivity. The present study describes the fabrication, calibration, and testing of simple, rugged, polymer-based sensors for detection of hydrazines in both ambient and vacuum environments. For reasonable choices of film thickness, initial resistance, and integration time, it is demonstrated that concentrations in the 0.1-100 ppb range can be monitored with an accuracy of +/ 20%. The sensor can be utilized for both dosimetric and real-time detection. Reproducible fabrication was achieved using standard spin-coating techniques. The polymer sensors exhibit good specificity to hydrazines in the presence of NH(3), amines, and ambient H(2)O and have a shelf-life of several years when stored in cold, dry conditions. PMID- 21619178 TI - Glucose-sensing electrode coated with polymer complex gel containing phenylboronic Acid. AB - We have prepared a copolymer containing both phenylboronic acid and tertiary amine moieties. The copolymer forms a stable complex with poly(vinyl alcohol) (PVA) since boronate moieties interact with PVA hydroxyl groups. The polymer polymer complex changes its swelling degree with glucose concentration in Dulbecco's phosphate-buffered saline solution (PBS) at pH 7.4, due to the higher complex formation of boronic acid moieties with glucose hydroxyl groups over those in PVA. Glucose-responsive swelling changes of a membrane complex were then utilized to control glucose-responsive current changes with a membrane-coated platinum electrode. Glucose addition to PBS induces swelling of the cast gel membrane, leading to increased diffusion of ion species and thus increased measurable current changes. Since the addition of methyl alpha-d-glucoside has little influence on the current changes, the current change by the addition of glucose is indicative of the high selectivity of this system for glucose and its cis-hydroxyl groups in glucose. It is observed that current changes are proportional to glucose concentration in the range 0-300 mg/dL. This range corresponds well to physiological blood glucose levels. Current change rates determined from the slope of the time course immediately after glucose addition are also proportional to glucose concentration within this range, yielding even higher sensitivity to the change in glucose concentration. Reproducible signal output is also demonstrated by repetitive, stepwise glucose concentration changes. These results support the applicability of the platinum electrode coated with the gel membrane complex comprising a phenylboronic acid-containing polymer and PVA for a novel glucose-sensing device. PMID- 21619179 TI - Voltammetric reduction of nickel and cobalt dimethylglyoximate. AB - The determination of cobalt and nickel in aqueous solutions by stripping voltammetry after adsorptive preconcentration is an established procedure. The method is highly sensitive, but there is some controversy concerning the reasons for the excellent sensitivity. Using a variety of voltammetric techniques, we have determined that the reduction of nickel dimethylglyoximate in an ammonia buffer is consistent with an overall process involving 16 or possibly 18 electrons. This hypothesis is confirmed by independently measuring the total quantity of metal adsorptively deposited on the mercury electrode and comparing the amount with the quantity of electricity required for its reduction. PMID- 21619180 TI - Accurate measurement of ruthenium isotopes by negative thermal ionization mass spectrometry. AB - The method for measurement of ruthenium isotopic composition as RuO(3)(-) by negative thermal ionization mass spectrometry (NTI-MS) is shown to be sensitive and accurate. Precise measurement of the (18)O/(16)O ratio, which is important for oxygen correction in NTI-MS, has also been made. Both Re and Pt filaments were tested, and the latter was proved to be more efficient for negative ion production. The mechanism of ion production with the addition of HI as a reducing reagent and Ba(NO(3))(2) as an ionizing enhancer was also studied. Sensitivity was found to be about 100 times higher than that of the positive mode. Factors related to negative ion formation are discussed, and parameters are optimized. The ionization efficiency has been improved to 0.7%. Ten nanograms of Ru yielded a total ion current of 3 * 10(-12) A for 1 h. The precisions of all Ru isotope ratios with a 100 ng sample size were better than 0.009%. PMID- 21619181 TI - Strategy for pulsed ionization methods on a sector mass spectrometer. AB - A method to help facilitate efficient implementation of pulsed ionization methods on a double-focusing sector mass spectrometer is described here. This method involves the addition of an inductive detector between the electric and magnetic sectors. The inductive detector will allow a crude, but complete time-of-flight mass spectrum to be acquired with as little as a single laser shot, thereby avoiding the necessity of sequentially compiling limited mass ranges as has been done previously. Mass analysis by the complete sector instrument can be performed simultaneously with the time-of-flight analysis. PMID- 21619182 TI - Tandem mass spectrometry of large biomolecule ions by blackbody infrared radiative dissociation. AB - A new method for the dissociation of large ions formed by electrospray ionization is demonstrated. Ions trapped in a Fourier transform mass spectrometer at pressures below 10(-)(8) Torr are dissociated by elevating the vacuum chamber to temperatures up to 215 degrees C. Rate constants for dissociation are measured and found to be independent of pressure below 10(-)(7) Torr. This indicates that the ions are activated by absorption of blackbody radiation emitted from the chamber walls. Dissociation efficiencies as high as 100% are obtained. There is no apparent mass limit to this method; ions as large as ubiquitin (8.6 kDa) are readily dissociated. Thermally stable ions, such as melittin 3+ (2.8 kDa), did not dissociate at temperatures up to 200 degrees C. This method is highly selective for low-energy fragmentation, from which limited sequence information can be obtained. From the temperature dependence of the dissociation rate constants, Arrhenius activation energies in the low-pressure limit are obtained. The lowest energy dissociation processes for the singly and doubly protonated ions of bradykinin are loss of NH(3) and formation of the b(2)/y(7) complementary pair, with activation energies of 1.3 and 0.8 eV, respectively. No loss of NH(3) is observed for the doubly protonated ion; some loss of H(2)O occurs. These results show that charge-charge interactions not only lower the activation energy for dissociation but also can dramatically change the fragmentation, most likely through changes in the gas-phase conformation of the ion. Dissociation of ubiquitin ions produces fragmentation similar to that obtained by IRMPD and SORI CAD. Higher charge state ions dissociate to produce y and b ions; the primary fragmentation process for low charge state ions is loss of H(2)O. PMID- 21619183 TI - Matrix-enhanced secondary ion mass spectrometry: a method for molecular analysis of solid surfaces. AB - A new methodology, matrix-enhanced secondary ion mass spectrometry (ME-SIMS), is reported for the molecular analysis of biomaterials. The technique applies static secondary ion mass spectrometry (SSIMS) techniques to samples prepared in a solid organic matrix similar to sample preparations used in matrix-assisted laser desorption/ionization (MALDI). Molecular ions are observed in this ion beam sputtering of organic mixtures for peptides and oligonucleotides up to masses on the order of 10 000 Da. This matrix-enhanced SIMS exhibits substantial increases in the ionization efficiency of selected analyte molecules compared to conventional SSIMS processes. Thus, higher mass peptides, proteins, and nucleic acids become accessible to near-surface analysis by ion beam techniques, and subpicomole sensitivity has been demonstrated. A number of matrices were examined for their efficiency in ME-SIMS applications, and these initial matrix studies focused on common MALDI matrices and their isomers. The results of this survey indicate that 2,5-dihydroxybenzoic acid provides the best general enhancement of molecular secondary ions emitted from analyte/matrix mixtures. PMID- 21619184 TI - Isotope ratio measurements with elemental-mode electrospray mass spectrometry. AB - The isotope ratio capabilities of an electrospray ionization source interfaced to a quadrupole mass spectrometer are described. With the instrument operated in the metal ion mode, isotope measurements of Ag, Tl, and Pb are conducted using elemental ions produced from 1 * 10(-)(4) M solutions of metal nitrates or acetates in methanol. For Ag and Tl, spray conditions are identified that produce spectra free of MH(+) ions. Unbiased Ag and Tl ratio measurements with precisions of ~0.2% RSD are readily attained. Further improvement in relative precision appears to be limited by temporal drift in the degree of mass discrimination imparted to the measurements by the mass spectrometer. Isotopic analysis of Pb is greatly complicated by significant yields of PbH(+) polyatomic ions. PMID- 21619185 TI - Theoretical description of the influence of external radial fields on the electroosmotic flow in capillary electrophoresis. AB - The effect of applying a radial voltage on the electroosmotic flow in capillary electrophoresis has been studied from a theoretical point of view. Based on Stern's model for the electric double layer on the surface of a fused silica capillary and on the Gouy-Chapman theory for the diffuse layer, equations describing the relation between the electroosmotic mobility and the radial electric field were derived. The thickness of the stagnant solution layer on the surface of the capillary, an important parameter in the calculations, was estimated from the electroosmotic mobility found in high-pH solutions. The theory developed predicts the experimental findings that the effect of the radial field levels off at high applied voltages and that it is smaller when the electroosmotic mobility without radial field is already high. The theoretical results were compared with experimental data taken from the literature. A good quantitative agreement was found. PMID- 21619186 TI - Effect of helium in helium headspace carbon dioxide cylinders on packed-column supercritical fluid chromatography. AB - Supercritical fluid chromatography of PAHs was performed with pure carbon dioxide and helium headspace carbon dioxide at various cylinder fill levels. The retention times of the PAHs increased when helium headspace carbon dioxide was used as a carrier fluid relative to pure carbon dioxide. The increased retention times were affected by the level of the liquid phase present in the helium headspace carbon dioxide cylinder. As more liquid phase was removed from the cylinder, the effect of helium on the solvating power of CO(2) was reduced because the relative amount of helium dissolved in the liquid phase decreased. Furthermore, the effect of helium headspace carbon dioxide was investigated with methanol-modified carbon dioxide mobile phases for the analysis of steroids. We observed that the relative solubility of helium in carbon dioxide resulted in longer retention times when compared to pure carbon dioxide as the liquid level of carbon dioxide decreased. PMID- 21619187 TI - Determination of atmospheric methyl bromide by cryotrapping-gas chromatography and application to soil kinetic studies using a dynamic dilution system. AB - Methyl bromide (CH(3)Br) is considered to be a major source of stratospheric Br, which contributes to the destruction of ozone. It is therefore necessary to understand the natural sinks of this compound and to accurately measure ambient mixing ratios. Methodology is described for the measurement of atmospheric CH(3)Br by cryotrapping-gas chromatography and its application to soil kinetics. A 2-propanol/dry ice cryotrap was used to preconcentrate CH(3)Br in standard and air samples, with subsequent detection using a gas chromatograph equipped with an O(2)-doped electron capture detector (GC-ECD). The GC-ECD cryotrapping method had a detection limit of 0.23 pmol of CH(3)Br. This is equivalent to the amount of CH(3)Br in a 500 mL sample of ambient air at the estimated northern hemisphere atmospheric mixing ratio of 11 parts per trillion by volume (pptv). A dynamic dilution system was developed to produce mixing ratios of CH(3)Br ranging between 4 and 1000 pptv. Calibrated mixing ratios of CH(3)Br produced with the dilution system were used to determine soil uptake kinetics employing a dynamic soil incubation method. PMID- 21619188 TI - Gas chromatographic isolation of individual compounds from complex matrices for radiocarbon dating. AB - This paper describes the application of a novel, practical approach for isolation of individual compounds from complex organic matrices for natural abundance radiocarbon measurement. This is achieved through the use of automated preparative capillary gas chromatography (PCGC) to separate and recover sufficient quantities of individual target compounds for (14)C analysis by accelerator mass spectrometry (AMS). We developed and tested this approach using a suite of samples (plant lipids, petroleums) whose ages spanned the (14)C time scale and which contained a variety of compound types (fatty acids, sterols, hydrocarbons). Comparison of individual compound and bulk radiocarbon signatures for the isotopically homogeneous samples studied revealed that Delta(14)C values generally agreed well (+/-10%). Background contamination was assessed at each stage of the isolation procedure, and incomplete solvent removal prior to combustion was the only significant source of additional carbon. Isotope fractionation was addressed through compound-specific stable carbon isotopic analyses. Fractionation of isotopes during isolation of individual compounds was minimal (<50/00 for delta(13)C), provided the entire peak was collected during PCGC. Trapping of partially coeluting peaks did cause errors, and these results highlight the importance of conducting stable carbon isotopic measurements of each trapped compound in concert with AMS for reliable radiocarbon measurements. The addition of carbon accompanying derivatization of functionalized compounds (e.g., fatty acids and sterols) prior to chromatographic separation represents a further source of potential error. This contribution can be removed using a simple isotopic mass balance approach. Based on these preliminary results, the PCGC-based approach holds promise for accurately determining (14)C ages on compounds specific to a given source within complex, heterogeneous samples. PMID- 21619189 TI - Selective vapor sorption by polymers and cavitands on acoustic wave sensors: is this molecular recognition? AB - Selectivity patterns for the sorption of organic vapors from the gas phase into cavitand monolayers on acoustic wave sensors are very similar to those seen for sorption of the same vapors by amorphous polymers, demonstrating that the vapor/cavitand selectivity patterns are determined primarily by solubility interactions. The amorphous polymers serve as controls demonstrating that the three-dimensional structure of a cavitand layer is not primarily responsible for the selectivity observed. Binding and selectivity in the examples cited are governed primarily by general dispersion interactions and not by specific oriented interactions that could lead to molecular recognition. PMID- 21619190 TI - Dielectric properties of a binary cryoelectrochemical solvent according to a solvatochromic probe. AB - The application of a solvatochromic dye to determination of the dielectric properties of organic solvent mixtures is described. The dye 2,6-diphenyl-4 (2,4,6-triphenylpyridino)phenolate (Reichardt's dye) features an intramolecular charge transfer band in its visible spectrum with a large associated change in dipole moment between the ground and excited states. The position of the charge transfer band is sensitive to local dielectric environment and so is an indicator of solvation of the dye. Room-temperature spectra of Reichardt's dye in the cryoelectrochemical solvent system 2:1 (v/v) ethyl chloride/butyronitrile and in the neat components are reported and related to the dielectric properties of the mixture. The effective dielectric properties of the mixture scale with the value of the Pekar factor, 1/epsilon(OP) - 1/epsilon(S), that is predicted from the mole fraction-weighted (or volume fraction-weighted) sum of the dielectric constants for the pure components, suggesting that strongly selective solvation of the dye by one component of the solvent mixture does not occur. The results are pertinent to a recent study of electrode kinetics in the solvent mixture. PMID- 21619191 TI - Determination of the quaternary ammonium surfactant ditallowdimethylammonium in digested sludges and marine sediments by supercritical fluid extraction and liquid chromatography with postcolumn ion-pair formation. AB - To study the phasing-out of the quaternary ammonium surfactant ditallowdimethylammonium cation (DTDMAC), concentrations of the cation in anaerobically stabilized sewage sludges were determined before and after its replacement by better degradable compounds. DTDMAC was quantitatively extracted from digested sludges using 380 atm of supercritical CO(2) modified with 30% methanol at 100 degrees C. Determination of DTDMAC was performed by normal-phase HPLC with postcolumn ion-pair formation and extraction with no sample cleanup. Mean concentrations of DTDMAC in sludges from five different municipal sewage treatment plants in Switzerland decreased from 3.67 g/kg (in 1991) to 0.96, 0.47, and 0.21 g/kg of dry sludge in 1992, 1993, and 1994, respectively. The precision of the method in digested sludge for 0.1-6.0 g/kg of dry matter, as indicated by the relative standard deviation, was typically 7%. The influence of the sample matrix was studied by performing supercritical fluid extraction (SFE) in coastal marine sediments. While SFE and a conventional liquid extraction method gave equal DTDMAC concentrations in sludges, the extraction of marine sediment samples yielded 30-40% higher DTDMAC values for SFE compared to those obtained by liquid extraction. The 94% drop in DTDMAC concentrations in digested sludges is due to the replacement of this substance and is a clear result of the producers' voluntary ban on its use in Europe. PMID- 21619192 TI - Characterization of Nonmicrobiological Paper Mill Deposits by Simultaneous TG-IR MS. AB - The problem of deposits and their identification is one of the most persistent ones facing paper mills. Materials other than fibers accumulate in closed water circulation systems and may appear as impurities in pulp and paper or can contaminate wires and felts or build up deposits at various points in the mills. Traditionally, chromatographic and spectroscopic methods have been employed in the analysis of deposits and other pulp and paper process impurities. Simultaneous thermogravimetry-infrared-mass spectrometric (TG-IR-MS) analysis is a relatively new technique which, when applied to the analysis of mill deposits, provides a rapid means of identifying the source of these contaminants. PMID- 21619193 TI - Qualitative identification of fumaric Acid and itaconic Acid in emulsion polymers. AB - A pyrolysis gas chromatography (Py-GC) technique has been used for the qualitative analysis of fumaric acid and itaconic acid as low-level monomers polymerized with other major monomers in emulsion polymers. In order for fumaric acid and itaconic acid to be detected through pyrolysis, the acids are derivatized with primary amines such as methylamine and ethylamine to form a cyclic imide. The detection of derivatized fumaric acid and itaconic acid is accomplished by atomic emission detection (AED). The structures of the derivatization-pyrolysis products have been elucidated by mass spectrometry. PMID- 21619194 TI - High-resolution capillary isoelectric focusing-electrospray ionization mass spectrometry for hemoglobin variants analysis. AB - On-line capillary isoelectric focusing (CIEF)-electrospray ionization mass spectrometry (ESIMS) as a two-dimensional separation system is employed for high resolution analysis of hemoglobin variants A, C, S, and F. The effects of moving ionic boundary inside the CIEF capillary and MS scan rate on the separation resolution and mass detection of hemoglobin variants are investigated. The formation of a moving ionic boundary due to the replacement of background electrolyte counterions with sheath liquid counterions can be minimized by combining cathodic mobilization with a gravity-induced hydrodynamic flow. Hemoglobin variants F and A, with a pI difference of 0.05 pH unit, are almost baseline resolved and identified in CIEF-ESIMS. The concentration detection limit for each hemoglobin variant is in the range of 10(-)(8) M, comparable to that obtained in two-dimensional gel electrophoresis using silver staining. Initial preconcentration during the focusing step and the use of single-ion monitoring scan mode are responsible for improving detection limits. PMID- 21619195 TI - Integrated on-capillary electrochemical detector for capillary electrophoresis. AB - An on-capillary electrochemical detector for capillary electrophoresis is described. It consists of a gold wire mounted permanently at the end of the capillary perpendicular to the direction of flow. This mode of detection eliminates the need for the micromanipulators or specially machined cell holders for alignment that are used for in-capillary detection modes. It also makes it possible to perform relatively fast CEEC separations using very short capillaries. The use of this detector for both off-column detection of catecholamines and end-column detection of carbohydrates by CE-PAD is described. PMID- 21619196 TI - Confocal Fluorescence Microscopic Imaging for Investigating the Analyte Distribution in MALDI Matrices. AB - The analytical performance of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry is strongly influenced by the method of analyte and matrix preparation. We report a nonintrusive method based on laser confocal microscopic imaging technology to examine the MALDI samples prepared by various protocols. In this method, the analyte is tagged with a fluorescent group. The matrix and analyte are prepared under the same conditions as those used in conventional MALDI experiments. It is demonstrated that confocal microscopy can provide clear, three-dimensional images of sample crystals as well as the analyte distribution within the crystals. It is shown that the analyte is incorporated into the matrix crystals for all the sample preparation protocols examined. Moreover, the confocal microscopic images reveal that, with the use of a dried-droplet method for sample/matrix preparation, the analyte is not uniformly distributed within the matrix crystals. In some crystals, no analyte is incorporated. In addition, it is found that large crystals formed using a slow growth process display a more uniform analyte distribution. Relatively more uniform analyte distribution is observed for samples prepared with the formation of microcrystals. The possible correlation between the ion signal variations observed in MALDI and the uniformity of the analyte distribution obtained by the confocal microscopic imaging method is discussed. Finally, a double-imaging method involving the use of two analytes with different labeling groups is demonstrated. It is found that different analytes are not coherently distributed in the matrix crystals. PMID- 21619197 TI - Continuous separation of high molecular weight compounds using a microliter volume free-flow electrophoresis microstructure. AB - A microliter volume free-flow electrophoresis microstructure (MU-FFE) was used to perform a continuous separation of high molecular weight compounds. The MU-FFE microstructure had a separation bed volume of 25 MUL and was fabricated from silicon using standard micromachining technology. Laser-induced fluorescence was used to detect the sample components, which were labeled with fluorescein isothiocyanate (FITC) prior to analysis. The continuous separation of human serum albumin (HSA), bradykinin, and ribonuclease A demonstrated that only 25 V/cm was required to isolate HSA from bradykinin and ribonuclease A, while 100 V/cm was needed for the separation of bradykinin from ribonuclease A. Comparison of the observed band broadening with the theoretical variance indicated that dispersion due to the initial bandwidth, diffusion, and hydrodynamic broadening were the main contributors to the band broadening of HSA and bradykinin. However, the band broadening for ribonuclease A could not be sufficiently accounted for using the above contributors. Adsorption was found to be a possible contributor to the overall variance for ribonuclease A. Calculation of the theoretical variance due to Joule heating indicated that broadening due to Joule heating effects was insignificant. This was likely due to the narrow cross-sectional area of the device, which facilitated efficient cooling. Electrohydrodynamic distortion was observed for HSA as it migrated toward the side bed. Studies of the resolution of bradykinin and ribonuclease A as a function of field strength at various sample and carrier flow rates indicated that, for maximum throughput, high field strengths and high flow rates were required. However, no optimal conditions were found. The MU-FFE device has a peak capacity of ~8 bands/cm, while for a typical separation of proteins using a commercial system, a peak capacity of 10 bands/cm is obtained. Thus, the resolving power of the MU-FFE device is similar to those of conventional systems. The continuous separation of tryptic digests of mellitin and cytochrome c demonstrated the ability to continuously separate more complex mixtures. Finally, modifications were made to the microstructure to facilitate fraction collection, and the fractionation of whole rat plasma was performed. Off line analysis of the resulting fractions indicated that the complete isolation of serum albumin and globulins was possible using a field strength of 25 V/cm. PMID- 21619198 TI - Head-Column Field-Amplified Sample Stacking in Binary System Capillary Electrophoresis: A Robust Approach Providing over 1000-Fold Sensitivity Enhancement. AB - Effective electrokinetic field-amplified sample injection occurring at the capillary inlet from a sample volume equivalent exceeding that of the capillary up to 10-fold is described and demonstrated to provide over 1000-fold sensitivity enhancement. Successful application of this head-column field-amplified sample stacking approach to the analysis of positively chargeable, hydrophobic compounds in binary system capillary electrophoresis is shown to require an initially introduced low-conductivity zone (water plug) of >1 mm length, a sample injection voltage <20 kV, and an injection time interval <60 s. Following these conditions for more than 1500 runs with capillaries of 50 MUm i.d. and about 20 cm effective length, damaging heat production during electroinjection within the low conductivity zone at the column inlet (boiling of solvent and possible deposition of solutes or fusing of capillary walls) could be prevented. The solute amount injected by head-column field-amplified sample stacking is further shown to be dependent on the organic fraction and the buffer in the sample solution. High content of organic solvent, low conductivity, and the presence of a small amount of H(+) (50-100 MUM) provides the highest sensitivity for analysis of positively chargeable model substances, including amiodarone and desethylamiodarone. Solutes present at the nanomolar level can thereby be accumulated from a sample volume equivalent of about 4 MUL (with injection of about 20 nL of sample solvent into the capillary) and measured by UV absorption detection. To prevent disturbances caused by electrolysis, sample vials should be employed only once. The data obtained further show that quantitation can be reliably performed using internal calibration based on peak height (RSDs for inter- and intraday determinations are on the 2% level). However, due to variation of the roughness of the capillary walls and cuts, the time interval between operational steps, and trace adsorption onto the capillary walls, the length of the water zone drawn by capillary action on the inlet side is not constant, and external calibration therefore cannot be employed for quantitation. PMID- 21619199 TI - Oligomeric separation of ionic and nonionic ethoxylated polymers by capillary gel electrophoresis. AB - Capillary gel electrophoresis (CGE) has proven itself as a superior, high resolution technique for separating proteins, peptides, oligonucleotides, and other naturally occurring molecules. In the years since its inception, few applications of CGE to nonbiological synthetic polymers have been reported. CGE has been applied to the separation of ionic and nonionic ethoxylated surfactants and poly(ethylene glycol) (PEG) oligomers. Oligomer distributions of several sulfated and phosphated alkylphenol ethoxylate surfactants have been baseline resolved with CGE on commercial cross-linked polyacrylamide gel columns. Nonionic surfactants and PEG oligomers were derivatized with phthalic anhydride in order to provide charge and detectability. PEG oligomers ranging from ethylene glycol to species containing more than 120 ethylene oxide units have been resolved. A linear relationship between migration time and molecular weight was found, which indicates that the separation mechanism is not simply based on molecular size but is also influenced by the electrophoretic mobility of the oligomers. The main drawbacks of CGE include relatively long analysis times and somewhat fragile and expensive columns. PMID- 21619200 TI - Capillary electrophoresis-fluorescence line-narrowing system for on-line structural characterization of molecular analytes. AB - We have demonstrated, for the first time, that capillary electrophoresis (CE) can be interfaced with low-temperature fluorescence line-narrowing (FLN) spectroscopy for on-line structural characterization. Detection by laser-induced fluorescence spectroscopy, under fluorescence non-line-narrowing and line-narrowing conditions, provides three-dimensional electropherograms and FLN spectra, which lead to significantly improved overall resolution and allow for structural characterization ("fingerprinting") of molecular analytes. This novel CE-FLN system consists of a modular CE system, instrumentation for FLN spectroscopy, and a specially designed capillary cryostat (CC). An absorbance detector serves to determine the migration rates of analytes. After the 77 K fluorescence-based electropherogram is generated, the temperature of the capillary is lowered to 4.2 K for high-resolution FLN characterization. Automated translation of the CC and capillary in the direction of the capillary axis allows the separated analytes to be sequentially characterized by fluorescence spectroscopy as the capillary is translated through the laser excitation region. Detection of fluorescence from stationary CE-separated analytes significantly improves the accuracy of quantitation and structural characterization. We believe that this interfacing represents an exciting addition to the rapidly evolving field of CE, providing a new and powerful tool for chemical analysis. The first application of the CE-FLN system to a mixture of polycyclic aromatic hydrocarbons is presented; prospects and future applications of CE-FLN are briefly addressed. PMID- 21619201 TI - Electrospray Ion Chromatography-Tandem Mass Spectrometry of Bromate at Sub-ppb Levels in Water. AB - An electrospray ion chromatography-tandem mass spectrometry (IC-MS/MS) method has been developed for the analysis of bromate ions in water. This IC-MS/MS method improves the limit of detection of bromate ions by a factor of 10. The method consists of solid phase extraction with an ion exchange column and elution of the analyte with water/methanol ammonium sulfate eluent on-line with a negative ion electrospray mass spectrometry detection. SPE requires sample pretreatment to remove any major ions that displace bromate, consisting of eliminating SO(4)(2)( ), Cl(-), and HCO(3)(-) ions respectively with barium-form, silver-form, and acid (H(+)-form) exchange resins. The methanolic sulfate eluent permits IC-MS coupling via an electrospray interface. BrO(3)(-) was selected in the first quadrupole (Q1) at two m/z values, 127 and 129, according to the isotope contributions of (79)Br and (81)Br. After fragmentation in the collision cell (second quadrupole, Q2), the third quadrupole (Q3) analyzes the product ions as (M - O)(-), (M - 2O)( ), and (M - 3O)(-). Among the six recordable transitions, four were selected, the other two yielding high background. A lowered resolution raised sensitivity by a factor of up to 3. The limit of quantitation of this method was 0.1 MUg/L. PMID- 21619202 TI - Sample preparation using a miniaturized supported liquid membrane device connected on-line to packed capillary liquid chromatography. AB - A miniaturized supported liquid membrane device has been developed for sample preparation and connected on-line to a packed capillary liquid chromatograph. The device consists of hydrophobic polypropylene hollow fiber, inserted and fastened in a cylindrical channel in a Kel-F piece. The pores of the fiber are filled with an organic solvent, in this study 6-undecanone, thus forming a liquid membrane. The sample is pumped on the outside of the hollow fiber (donor), and the analytes are selectively enriched and trapped in the fiber lumen (acceptor). With this approach, the volume of the acceptor solution can be kept as low as 1-2 MUL. This stagnant acceptor solution is then transferred through capillaries attached to the fiber ends to the LC system. The system was tested with a secondary amine (bambuterol), as a model substance in aqueous standard solutions as well as in plasma. The best extraction efficiency in aqueous solution, with an acceptor volume of 1.9 MUL, was 32.5% at a donor flow rate of 2.5 MUL/min. At flow rates above 20 MUL/min, the concentration enrichment per time unit was approximately constant, at 0.9 times/min, i.e., 9 times enrichment in about 10 min. The overall repeatability (RSD) for spiked plasma samples was ~4% (n = 12). Linear calibration curves of peak area versus bambuterol concentration were obtained for both aqueous standard solutions and spiked plasma samples. The detection limit for bambuterol in plasma, after 10 min of extraction at a flow rate of 24 MUL/min, was 80 nM. PMID- 21619203 TI - Measurement of Internal Acyl Migration Reaction Kinetics Using Directly Coupled HPLC-NMR: Application for the Positional Isomers of Synthetic (2-Fluorobenzoyl) d-glucopyranuronic Acid. AB - Ester glucuronides (1-O-acyl-beta-d-glucopyranuronates) of many drugs may undergo internal acyl migration reactions, resulting in the formation of new positional isomers with both alpha- and beta-anomers. We illustrate here a novel approach for the direct investigation of the acyl migration kinetics of ester glucuronides and show the application with respect to the isomers of synthetic (2 fluorobenzoyl)-d-glucopyranuronic acid. Individual isomers were separated from an equilibrium mixture containing the beta-1-O-acyl, alpha- and beta-2-O-acyl, alpha and beta-3-O-acyl, and alpha- and beta-4-O-acyl isomers at pH 7.4 in 20 mM phosphate buffer. The interconverting isomers were separated using reversed-phase HPLC and pumped directly into a dedicated on-line NMR flow probe in a 600 MHz NMR spectrometer. The flow was stopped with each isomer in the NMR flow probe, and sequential NMR spectra were collected at 25 degrees C, allowing direct measurement of the production of positional isomers from each selectively isolated glucuronide isomer. All of the positional isomers and anomers were characterized, and relative quantities determined, and a kinetic model describing the rearrangement reactions was constructed. The acyl migration reaction kinetics were simulated using a theoretical approach using nine first-order rate constants determined for the acyl migration reactions and six first-order rate constants describing the mutarotation each of the 2-, 3-, and 4-positional isomers. The rate constants (in h(-)(1)) for the rearrangement reactions of the 2 fluorobenzoyl glucuronide isomers were as follows: beta-1-O-acyl, 0.29 +/- 0.01; alpha-2-O-acyl, 0.11 +/- 0.01; beta-2-O-acyl, 0.07 +/- 0.01; alpha-3-O-acyl, 0.10 +/- 0.01; beta-3-O-acyl, 0.09 +/- 0.01; alpha-4-O-acyl, 0.09 +/- 0.01; and beta-4 O-acyl, 0.06 +/- 0.01. The alpha- and beta-anomerization rates were estimated on the basis of the kinetics model; the anomerization rates of the 4-O-acyl isomers were additionally determined experimentally using directly coupled HPLC-NMR. The fitted anomerization rates for the 4-O-acyl isomer were 0.80 (alpha -> beta) and 0.50 h(-)(1) (beta -> alpha), whereas the experimentally estimated anomerization rates were 0.89 +/- 0.1 and 0.52 +/- 0.1 h(-)(1), respectively. The dynamic stop flow HPLC-NMR approach allows unique kinetic information to be obtained relating to glucuronide reactivity, and this approach will be useful in future structure reactivity studies on drug ester glucuronides and their properties. PMID- 21619204 TI - Classical retention mechanism in ion exchange chromatography. Theory and experiment. AB - The classical model for ion exchange chromatography is characterized by firmly adsorbed driving ions at the surface of the stationary phase in an amount required by electroneutrality and stoichiometric ion exchange between the bulk of the eluent electrolyte and this immobilized Stern layer. Retention equations have been derived for system peaks, labeled eluent ions, and analytes in a system containing only strong electrolytes by strictly respecting this model. It is shown that the classical model described dependence of retention data on concentration and composition of the binary eluent with excellent precision, but the resulting system parameters were not self-consistent. Inconsistency of the results might be due to contributions from another retention mechanism. PMID- 21619205 TI - Mass sensitivity of the thin-rod acoustic wave sensor operated in flexural and extensional modes. AB - The mass sensitivities of the thin-rod acoustic wave sensor in both flexural and extensional acoustic modes are presented. These are based on experiments involving the electrodeposition of a test loading material onto a thin metal fiber (the thin rod) in a delay line configuration. Only small changes in acoustic loss occur when the device is immersed in an electrolyte, particularly in the flexural mode. Copper and lead have been used as test materials to confirm that the effects of elasticity can give rise to positive and negative mass sensitivities, respectively. The experimental and theoretical values all agree in sign and are of the same order of magnitude. This result confirms a refined theoretical model that includes incorporation of the effects of elasticity and inertia. An increase in experimental mass sensitivity with decrease in fiber radius is one of the advantages for the construction of a sensitive chemical sensor based on the thin-rod device. The sensor can be operated with facility in both gas and liquid phases and offers a new technique for the study of interfacial electrochemistry on metal surfaces. The phenomenon of mechanical resonance was observed during a number of experiments. PMID- 21619206 TI - Ionization constants of ginkgolide B in aqueous solution. AB - The thermodynamic ionization constants (pK(a)(1), pK(a)(2), and pK(a)(3)) of ginkgolide B (9H-1,7a-(epoxymethano)-1H,6aH-cyclopenta[c]furo[2,3-b]furo [3',2':3,4]cyclopenta[1,2-d]furan-5,9,12-(4H)-trione, 3-tert-butylhexahydro 4,7b,11-trihydroxy-8-methyl-) in aqueous solution have been settled by pH-metric and NMR studies. The three macroscopic pK(a) values as well as the water solubility and the water/n-octanol partition coefficient have been extracted from pH-metric data by means of a nonlinear regression methodology. NMR spectroscopy provided confirmation of the values of the macroscopic constants, information about the effective ionization pathways, and an estimation of the proportions of the various forms under physiologically relevant conditions. PMID- 21619207 TI - Oxygen sensors based on luminescence quenching of metal complexes: osmium complexes suitable for laser diode excitation. AB - Oxygen quenching of a series of Os(II) complexes with alpha-diimine ligands has been studied in a predominantly poly(dimethylsiloxane) (PDMS) polymer and in Gp 163 (an acrylate modified PDMS). Unlike previous Ru(II) complexes used as oxygen sensors, the Os complexes can be excited by readily available, high-intensity, low-cost, red diode lasers at 635, 650, and 670 nm. Variations in the polymer properties have been made in order to delineate the structural features important for satisfactory use of supports for oxygen sensors. A key factor is matching the hydrophobicity of the sensor and support for optimal compatibility and minimizing the size of low oxygen diffusion domains. PMID- 21619208 TI - Atmospheric pressure microwave sample preparation procedure for the combined analysis of total phosphorus and kjeldahl nitrogen. AB - An atmospheric pressure microwave digestion method has been developed for the combined analysis of total phosphorus and Kjeldahl nitrogen in complex matrices. In comparison to the digestion steps in EPA Methods 365.4 (total phosphorus) and 351.x (Kjeldahl nitrogen), this method requires less time, eliminates the need for a catalyst, and reduces the toxicity of the waste significantly. It employs a microwave-assisted digestion step, using refluxing borosilicate glass vessels at atmospheric pressure. Traditionally, this method has a time-consuming sample preparation step and generates toxic waste through the use of heavy metal catalysts. These advantages are gained by the combination of a high boiling point acid (sulfuric acid) and the application of focused microwave irradiation, which enhances the digestion process by direct energy coupling. NIST standard reference materials 1572 (citrus leaves), 1577a (bovine liver), and 1566 (oyster tissue) and tryptophan were analyzed to validate the method. Phosphorus concentrations were determined by the colorimetric ascorbic acid method outlined in EPA Method 365.3. Kjeldahl nitrogen concentrations were determined using EPA Method 351.1. The results of the analyses showed good precision and are in excellent agreement with the NIST published values for both elements. PMID- 21619209 TI - Effects of polymer matrices on calibration functions of luminescent oxygen sensors based on porphyrin ketone complexes. AB - The design of luminescent oxygen sensors is guided by optimizing sensitivity and/or the form of the calibration function. Both qualities are governed by the molecular processes of luminescence quenching. To evaluate the influence of matrix effects, we prepared membranes based on oxygen-sensitive phosphorescent complexes of porphyrin ketones dissolved in plasticizer-free poly(vinyl chloride) (PVC) and polystyrene (PS). In a PVC matrix, both platinum(II) and palladium(II) octaethylporphyrin ketones exhibited perfectly linear Stern-Volmer intensity plots and almost single-exponential excited state decays. In a PS matrix, the sensitivity of palladium(II) octaethylporphyrin ketone was among the highest reported to date. Yet, slightly nonlinear Stern-Volmer plots and nonexponential decays illustrate the significance of matrix effects of PS. Addition of plasticizers to PVC-based sensors allowed tuning of the oxygen sensitivity in a wide range, while the Stern-Volmer plots became pronouncedly nonlinear. For the plasticizer bis(2-ethylhexyl) adipate, the decay profile was single-exponential in the absence but nonexponential in the presence of oxygen, which is attributed to a distribution of quenching rate constants. PMID- 21619210 TI - A glucose sensor made of an enzymatic clay-modified electrode and methyl viologen mediator. AB - A novel glucose sensor has been contrived by immobilizing glucose oxidase between two nontronite clay coatings on glassy carbon electrode with methyl viologen as mediator. The sandwich configuration proved to be very effective in the determination of glucose. The response of the glucose sensor was determined by measuring cyclic voltammetric peak current values under aerobic solution conditions. The effects of the amount of enzyme immobilized, the operating pH, and the common interferences on the response of the glucose sensor were studied. The detection limit was 5 MUM, with a linear range extending to about 6 mM, giving a dynamic range of over 3 orders of magnitude for 0.1 mM methyl viologen. When stored in pH 7 phosphate buffer at 4 degrees C, the sensor shows almost no change in performance after operating for at least 2 months. A mechanism for the operation of the glucose sensor is also proposed. PMID- 21619211 TI - Electrogenerated chemiluminescence. 58. Ligand-sensitized electrogenerated chemiluminescence in europium labels. AB - The electrochemistry and electrogenerated chemiluminescence (ECL) of a series of europium chelates, cryptates, and mixed-ligand chelate/cryptand complexes were studied. The complexes were of the following general forms: EuL(4)(-), where L = beta-diketonate, a bis-chelating ligand (such as dibenzoylmethide), added as salts (A)EuL(4), where A = tetrabutylammonium ion or piperidinium ion (pipH(+)); Eu(crypt)(3+), where crypt = a cryptand ligand, e.g., 4,7,13,16,21-pentaoxa-1,10 diazabicyclo[8,8,5]tricosane; and Eu(crypt)(L)(2+) for the mixed-ligand systems. ECL was obtained for the chelates and mixed-ligand systems by reducing the complexes at a Pt electrode in the presence of peroxydisulfate in acetonitrile solutions and was attributed to the electron-transfer reaction between the reduced bound ligands and SO(4)(*)(-), followed by intramolecular excitation transfer from the excited ligand orbitals to the metal-centered 4f states. No ECL was observed under the same conditions for the europium complexes incorporating only the cryptand ligands in aqueous solution. The ECL spectra matched the photoluminescence spectra with a narrow emission band observed at 612 nm, corresponding to a metal-centered 4f-4f transition. The ECL efficiencies for the ECL-active species were low, about 10(-)(1)-10(-)(4)% of that of the Ru(bpy)(3)(2+)/S(2)O(8)(2)(-) system under similar conditions. PMID- 21619212 TI - Scanning electrochemical microscopy. 32. Gallium ultramicroelectrodes and their application in ion-selective probes. AB - Gallium ultramicroelectrodes for amperometric measurements in scanning electrochemical microscopy were fabricated by introduction of liquid Ga into drawn glass micropipets. Cyclic voltammetry of Ru(NH(3))(6)(3+) and the use of this species for SECM imaging is described. Double-barrel micropipet tips with a Ga amperometric electrode and an ion-selective (K(+)) potentiometric probe can also be constructed. This probe was used to image the K(+) activity near a 20-MUm diameter lumen of a glass capillary. PMID- 21619213 TI - Two-dimensional model of a direct current glow discharge: description of the argon metastable atoms, sputtered atoms, and ions. AB - A two-dimensional model is presented that describes the behavior of argon metastable atoms, copper atoms, and copper ions in an argon direct current glow discharge, in the standard cell of the VG9000 glow discharge mass spectrometer for analyzing flat samples. The model is combined with a previously developed model for the electrons, argon ions, and atoms in the same cell to obtain an overall picture of the glow discharge. The results of the present model comprise the number densities of the described plasma species, the relative contributions of different production and loss processes for the argon metastable atoms, the thermalization profile of the sputtered copper atoms, the relative importance of the different ionization mechanisms for the copper atoms, the ionization degree of copper, the copper ion-to-argon ion density ratio, and the relative roles of copper ions, argon ions, and atoms in the sputtering process. All these quantities are calculated for a range of voltages and pressures. Moreover, since the sticking coefficient of copper atoms on solid surfaces is not well-known in the literature, the influence of this parameter on the results is briefly discussed. PMID- 21619214 TI - Improved identification of noisy spectra using higher-ordered correlation spectral analysis. AB - We used a higher-order correlation-based method of comparison for spectral identification. Higher-order correlations are an extension of the more familiar second-order cross-correlation function and have the significant advantage of being theoretically shown to eliminate noise of unknown spectral density under certain conditions. Specifically, we applied a third-order correlation technique to the identification of similar IR spectra in the presence of noise. We were able to reduce the effects of noise from a second-order correlation measurement by further processing the measurement with a third-order autocorrelation. Our results showed that the third-order correlation-based method increased the probability of detection of a spectrum in the presence of noise, when compared to using a second-order technique alone. The probability of detection increased enough at low signal-to-noise ratios that this technique may be useful when a second-order correlation technique is not acceptable. The third-order technique is applicable to a single experiment, but improved results were found by averaging the results of multiple experiments. PMID- 21619215 TI - A capillary array gel electrophoresis system using multiple laser focusing for DNA sequencing. AB - A very simple and highly sensitive capillary array gel electrophoresis system is constructed to analyze DNA fragments. On-column detection of DNA migration in a large number of gel-filled capillaries is carried out using side-entry laser irradiation and with a CCD camera, although it has been considered impossible because the irradiation laser is scattered strongly at the surfaces of the first few capillaries. By optimizing optical conditions, the laser beam can be focused repeatedly to irradiate all the capillaries held on a plate by working each capillary as a cylindrical convex lens. DNA sequencing samples migrating in 24 capillaries can simultaneously be analyzed with the system. PMID- 21619216 TI - Screen-printable sol-gel enzyme-containing carbon inks. AB - Enzymes usually cannot withstand the high-temperature curing associated with the thick-film fabrication process and require a separate immobilization step in connection with the production of single-use biosensors. We report on the development of sol-gel-derived enzyme-containing carbon inks that display compatibility with the screen-printing process. Such coupling of sol-gel and thick-film technologies offers a one-step fabrication of disposable enzyme electrodes, as it obviates the need for thermal curing. The enzyme-containing sol gel carbon ink, prepared by dispersing the biocatalyst, along with the graphite powder and a binder, within the sol-gel precursors, is cured very rapidly (10 min) at low temperature (4 degrees C). The influence of the ink preparation conditions is explored, and the sensor performance is evaluated in connection with the incorporation of glucose oxidase or horseradish peroxidase. The resulting strips are stable for at least 3 months. Such sol-gel-derived carbon inks should serve as hosts for other heat-sensitive biomaterials in connection with the microfabrication of various thick-film biosensors. PMID- 21619217 TI - Silica xerogel as a continuous column support for high-performance liquid chromatography. AB - A preliminary study of the chromatographic performance and permeability of a continuous silica xerogel column under reversed-phase HPLC conditions was performed. A porous chromatographic support was synthesized inside a 0.32 mm i.d. * 13 cm fused silica tube from potassium silicate solution and derivatized with dimethyloctadecylchlorosilane. The plate height at 0.01 cm/s (0.5 MUL/min), near the apparent optimum linear velocity, was about 65 MUm. The column efficiencies in terms of numbers of plates per meter were 5000 and 13 000 for ethyl benzoate (k = 0.8) and naphthalene (k = 2.0), respectively, at 0.5 MUL/min. The major parameter affecting column efficiency was the heterogeneous morphology of the xerogel, modifications to which are expected to improve chromatographic performance. The column provided efficiencies comparable to those reported for continuous polymeric columns but less than that previously reported for a continuous silica column. Gradient elution mode was demonstrated with a mixture of polycyclic aromatic hydrocarbons. The column was highly permeable, exhibiting a linear dependence of pressure to flow rate and a back pressure of only 632 psi at 10 MUL/min when a 95% aqueous mobile phase was used. PMID- 21619218 TI - Solenoid pumps for flow injection analysis. AB - Methods employing flow injection analysis (FIA), particularly for in situ seawater techniques, would benefit from reduction in pump size and power requirement, longer maintenance intervals, and the ability to incorporate microprocessor control of each reagent and sample flow stream. In this work, the peristaltic pump of a conventional FIA system was replaced by three solenoid driven diaphragm pumps with integral Viton check valves, and the system was tested by performing the simple nitrite analysis, which has well-defined FIA performance characteristics. Sixty injections per hour were possible with flow rates of 0.5 mL/min for reagents and sample. The coefficient of variation was 1% for 10 MUM NO(2)(-) concentrations, and the detection limit was less than 0.1 MUM NO(2)(-). These values match the reported performance for this method using peristaltic pumps. PMID- 21619219 TI - Maximum entropy method for frequency domain fluorescence lifetime analysis. 1. Effects of frequency range and random noise. AB - The maximum entropy method (MEM) provides a self-modeling fit to data in which minimization of the chi(2) goodness-of-fit parameter is coupled with maximization of a statistical entropy function. We have found that MEM provides an excellent visual description of the uncertainties, errors, and limitations associated with the distributions which it recovers. To more accurately interpret fluorescence lifetime distributions recovered by the MEM from frequency domain lifetime data, a detailed examination of the effects of frequency range, noise, data set size, and sample heterogeneity was carried out for both simulated and real data. Results clearly demonstrate that the frequency range in which data are collected can affect the number and nature of the fluorescence lifetime components that are recovered by MEM, and the quality of the data at the frequencies that are optimal for a given lifetime is also crucial. Expansion of sufficient data sets to include more frequencies, or more replicates at the same frequencies, provides little improvement over the original data set when the lifetimes are well windowed by the frequency range. Synergism among multiple components in a sample can affect the recovered distribution, by shifting and splitting poorly windowed components and broadening the recovered peaks for all components. These effects are related to the number of components for which evidence must be found. PMID- 21619220 TI - Analysis of Gasolines by FT-IR Spectroscopy. AB - An experimental and computational protocol was established for the simultaneous determination of several key gasoline properties from a single Fourier transform infrared (FT-IR) spectrum. The study has shown that midband FT-IR spectroscopy combined with multivariate calibration analysis is a versatile, efficient, and accurate technique for the simultaneous estimation of key gasoline properties within about 1 min with less than 2 mL of sample. The FT-IR-derived values of gasoline properties include research and motor octane numbers, aromatic, olefinic, and saturated hydrocarbon content, benzene content, and concentrations of ethanol, methyl tert-butyl ether, and total oxygen. Concentrations of other oxygenated compounds are expected to be equally predictable. However, since these oxygen-containing species have not been adequately represented among the currently commercially available gasoline samples, their calibration may only be achieved using laboratory fuel blends. Midrange boiling point data may also be estimated. Fuel properties determined by minor concentrations of fuel components, e.g., flash point, sulfur content, etc., may not be modeled because the corresponding FT-IR signals are below detection limits of presented experimental protocol. The precision of this procedure was shown to be comparable to reproducibility of the standard laboratory analyses used for direct measurement of specific fuel properties, with squared correlation coefficient (R(2)) ranging from 0.94 to 0.99 between the two sets of measurements. This new methodology could increase the corresponding output of the petroleum laboratories by a factor of over 200 to 1 while maintaining data integrity and minimizing sample requirements, environmental hazards, and cost. PMID- 21619221 TI - Resolution and chemical formula identification of aromatic hydrocarbons and aromatic compounds containing sulfur, nitrogen, or oxygen in petroleum distillates and refinery streams. AB - An all-glass heated inlet system has been interfaced to a dual-trap Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometer. The inlet vaporizes a mixture of species of widely different boiling points, and the interface maintains a large (factor of 10(10)) pressure gradient between the inlet and the mass spectrometer, making possible the analysis of petroleum distillates and refinery streams at very high mass resolution. Ions generated by low-energy electron ionization in the source trap of the spectrometer are transferred to the analyzer trap, where the pressure is at least 2 orders of magnitude lower. Singly charged ions from a mass window of ~20 u are isolated by stored-waveform radial ejection, to reduce space charge and increase digital resolution: routine mass resolving power >200 000 (based on magnitude-mode peak full width at half-height) is thereby achieved throughout the full mass window. The mass window may be incremented stepwise to cover the full mass range of several hundred units. The FT-ICR mass spectrum of a gas oil aromatic neutral fraction contained peaks resulting from the resolution of ions having 358 distinct formulas over a mass range of ~42 u. C(3)/SH(4), (13)C/CH, (13)CH/N, CH(2)/N, and other mass doublets were baseline-resolved, yielding typical mass measurement inaccuracies of ~1 ppm. For example, (13)C(12)C(17)H(20)S(+) and C(21)H(17)(+), which differ by only 0.0011 u at ~269 u, were clearly resolved. A 40 000 resolving power low-voltage spectrum of the aromatic neutrals, acquired by use of a Kratos MS-50 double focusing instrument, was processed with a computer-based deisotoping/formula assignment procedure. The algorithm of the program is outlined and illustrated. Remarkably good agreement exists between the FT-ICR and MS-50 results. However, instrumental rather than indirect resolution of ions clearly enhances analytical accuracy and significantly reduces data-processing time. Thus, we have demonstrated that FT-ICR is the mass analysis of choice for differentiating hydrocarbons from heteroatom-containing compounds in petroleum distillates and refinery streams. PMID- 21619222 TI - Orthogonal projection approach applied to peak purity assessment. AB - The orthogonal projection approach (OPA), a stepwise approach based on an orthogonalization algorithm, is proposed. The performance of OPA for the assessment of peak purity in HPLC-DAD is described and compared with that of SIMPLISMA. The occurrence of artifacts in both approaches under nonideal situations is discussed. PMID- 21619223 TI - Formation and verification of the structure of the 1-fluorenylmethyl chloroformate derivative of sulfamethazine. AB - Sulfamethazine (SMZ) is derivatized with 1-fluorenylmethyl chloroformate (FMOC) to form the fluorescent adduct SMZ-FMOC. Conditions for formation are optimized with respect to pH, reagent concentration, and reagent ratio. Reagent and product profiles (including the hydrolysis byproduct FMOC-OH) versus time are followed by reversed phase HPLC with UV absorbance detection. FMOC-SMZ has been crystallized, its composition confirmed by microanalysis, and its structure corroborated by IR and NMR spectroscopy. From 10 down to 1 ppm, there is clear gentle curvature in the fluorescence intensity of SMZ-FMOC. The linear response range extends from above 100 ppb down to about 100 ppt, and an increase in sensitivity for the fluorescent detection of FMOC-SMZ (over the usual UV absorbance detection of SMZ) is calculated to be better than 3 orders of magnitude. PMID- 21619224 TI - Design and evaluation of a new thermospray liquid/liquid extractor for the extraction of semivolatile and nonvolatile organic compounds from water. AB - The recovery of several semivolatile organic compounds (SVOCs) using a new thermospray liquid/liquid extractor (TSLLE) was investigated. The base system includes a 300 mL multiport extraction vessel, jacketed in a 500 mL cooling flask, a dual-stage condenser for progressive cooling, several thermospray probes, and solvent/sample delivery systems. Aqueous mixtures of SVOCs were used to evaluate the TSLLE. For most compounds, recovery values of 80-100% were obtained during a single cycle in <1 h. The design, evaluation, and extraction capability of the TSLLE are discussed. PMID- 21619225 TI - Temperature-Responsive Chromatography Using Poly(N-isopropylacrylamide)-Modified Silica. AB - A new concept in chromatography is proposed that utilizes a temperature responsive surface with a constant aqueous mobile phase. The surface of the silica stationary phase in high-performance liquid chromatography (HPLC) has been modified with temperature-responsive polymers to exhibit temperature-controlled hydrophilic/hydrophobic changes. Poly(N-isopropylacrylamide) (PIPAAm) was grafted onto (aminopropyl)silica using an activated ester-amine coupling method. These grafted silica surfaces show hydrophilic properties at lower temperatures which, as temperature increases, transform to hydrophobic surface properties. The elution profile of five mixed steroids on an HPLC column packed with this material depends largely on the temperature of the aqueous mobile phase. Retention times increase with increasing temperature without any change in the eluent. Changes in the retention times of hydrophobic steroids were larger than those for hydrophilic steroids. The temperature-responsive interaction between PIPAAm-modified silica and these steroids is proposed to result from changes in the surface properties of the HPLC stationary phase by the transition of hydrophilic/hydrophobic surface-grafted IPAAm polymers. We demonstrate a novel and useful new chromatography system in which surface properties and the resulting function of the HPLC stationary phase are controlled by external temperature changes. This method should be effective in biological and biomedical separations of peptides and proteins using only aqueous mobile phases. PMID- 21619226 TI - Application of molecular imprinting to the development of aqueous buffer and organic solvent based radioligand binding assays for (s)-propranolol. AB - Antibody mimics have been prepared by molecular imprinting of (S)-propranolol and applied in the development of radioligand binding assays for the imprint species. In the assays, polymer particles, radioligand, and analyte were incubated either in an organic solvent or an aqueous buffer, with similarly high sensitivity under both set of conditions. Optimization of the assay conditions led to 100-1000-fold improvements in the limit of determination and 100-1000-fold reductions of the amount of imprinted polymer used, compared with previous studies of this novel type of assay. The present assay uses only 10-50 MUg of polymer, and the limit of determination is about 6 nM. The toluene-based assay showed excellent enantioselectivity, the cross-reactivity of the R enantiomer being only 1%, which is better than that demonstrated by biological antibodies. The aqueous buffer based assay showed high substrate selectivity for propranolol in the presence of structurally similar beta-blockers. The different selectivity profiles obtained are due to a different balance between hydrophobic and polar interactions in toluene and water, since polar interactions, such as hydrogen bonds, are strong in apolar solvents and hydrophobic interactions are strong in water. PMID- 21619227 TI - Wetting of octadecylsilylated silica in methanol-water eluents. AB - The wetting of an octadecylsilylated silica in methanol-water mixtures was studied by optical transmittance, visual observations, and measurements of the retention of model compounds. The octadecylsilylated silica particles remain wetted as the methanol content is decreased from 100% (v/v) to 20% (v/v). With the methanol content in the range of 20% (v/v) to 10% (v/v), the octadecylsilylated silica particles are still wetted, but the degree of solvation of the C(18) chains decreases with decreasing methanol concentration. The octadecylsilylated silica particles are not wetted when the methanol content in the mobile phase is lower than 10% (v/v). After equilibration with water, the octadecylsilylated silica particles remain nonwetted until the methanol content reaches 65% (v/v). The nonwetted phase showed significantly reduced chromatographic retention. A much longer equilibration time is required when the octadecylsilylated silica particles are not well solvated or nonwetted. The retentive behavior of the column will depend on the history of mobile-phase exposure. PMID- 21619228 TI - Solid-phase microextraction as a method for estimating the octanol-water partition coefficient. AB - The determination of octanol-water partition coefficients (log K(ow)) is important for the prediction of the fate of organic pollutants in the environment. Traditionally, log K(ow) values are determined by shake-flask, estimated by, e.g., HPLC retention data, or calculated, e.g., from ClogP. In this paper, an alternative approach is reported that allows log K(ow) to be estimated from solid-phase microextraction (SPME) data. Previously reported attempts to correlate SPME data with log K(ow) are discussed. The results obtained in this work for six phenols, using an 85 MUm polyacrylate-coated fiber, indicate that SPME is a viable method for estimating log K(ow) values <3.5. PMID- 21619229 TI - Affinity detection of low molecular weight analytes. AB - In this paper we report attempts to detect directly the binding of a low molecular weight substance to a protein binding site. An optical transducer based on reflectometric interference spectroscopy (RIFS) was used to detect the binding of biotin (244 g/mol) to a thin silica film surface coated with streptavidin. RIFS allows measurement of changes in the optical thickness of thin transparent films with high resolution. During immobilization of streptavidin, an increase in layer thickness of about 5 nm was detected. Subsequent incubation with biotin (4 MUM) resulted in a thickness increase of about 70 pm. Repeated incubation with biotin gave no further increase in layer thickness. The lowest biotin concentration showing significant effects was 40 nM. Incubation with benzoic acid (40 MUM) gave no thickness change. The setup allowed significant detection of thickness increases of 2 pm and above. Therefore, the thickness effects observed in the study could be unambiguously and clearly identified. PMID- 21619230 TI - Quantitative analysis of fuel-related hydrocarbons in surface water and wastewater samples by solid-phase microextraction. AB - Solid-phase microextraction (SPME) parameters were examined on water contaminated with hydrocarbons including benzene and alkylbenzenes, n-alkanes, and polycyclic aromatic hydrocarbons (PAHs). Absorption equilibration times ranged from several minutes for low molecular weight compounds such as benzene to 5 h for high molecular weight compounds such as benzo[a]pyrene. Under equilibrium conditions, SPME analysis with GC/FID was linear over 3-6 orders of magnitude, with linear correlation coefficients (r(2)) greater than 0.96. Experimentally determined FID detection limits ranged from ~30 ppt (w/w hydrocarbon/sample water) for high molecular weight PAHs (e.g., MW > 202) to ~1 ppb for low molecular weight aromatic hydrocarbons. Experimental distribution constants (K) were different with 100- and 7-MUm poly(dimethylsiloxane) fibers, and poor correlations with previously published values suggest that K depends on the fiber coating thickness and the sorbent preparation method. The sensitivity of SPME analysis is not significantly enhanced by larger sample volumes, since increasing the water volume (e.g., from 1 to 100 mL) has little effect on the number of analyte molecules absorbed by the fiber, especially for compounds with K < 500. Water sample storage should utilize silanized glassware, since hydrocarbon losses up to 70% could be attributed to unsilanized glassware walls when samples were stored for 48 h. Hydrocarbon losses at part-per-billion concentrations also occurred with surface waters due to partitioning onto part-per-thousand concentrations of suspended solids. Quantitative determinations of aromatic and aliphatic hydrocarbons (e.g., in gasoline-contaminated water) can be performed using GC/MS with deuterated internal standard or standard addition calibration as long as the target components or standards had unique ions for quantitation or sufficient chromatographic resolution from interferences. SPME analysis gave good quantitative performance with surface waters having high suspended sediment contents, as well as with coal gasification wastewater which contained matrix organics at 10(6)-fold higher concentrations than the target aromatic hydrocarbons. Good agreement was obtained between a 45-min SPME and methylene chloride extraction for the determination of PAH concentrations in creosote contaminated water, demonstrating that SPME is a useful technique for the rapid determination of hydrocarbons in complex water matrices. PMID- 21619231 TI - Determination of total organic emissions from hazardous waste combustors. AB - U.S. Environmental Protection Agency risk assessment guidance currently under development for evaluation of permitting information from hazardous waste combustors requires a quantity referred to as "total organic carbon". The risk guidance does not define this term precisely, nor does it explain how it should be determined. This paper discusses basic principles of sampling and analysis of stack emissions for "total organics", best currently available technology, and the status of two ongoing projects designed to provide guidance and to improve analysis procedures. Determination of total organics from stack emissions is much more complicated than might be expected, and more published guidance is badly needed. The best scheme available for analysis of stack emissions for total organics to be used in material balance style "bookkeeping" includes determination of organics content in three boiling point ranges: <100 degrees C, 100 degrees C-300 degrees C, and >300 degrees C. Total organic carbon is not a useful quantity, since it includes soot, polymeric material, and other nonextractable organic materials. Total organics has been found to be an imperfect but less misleading term. Various calculations can be made and conclusions can be drawn on the basis of the contents of the individual boiling point ranges, as determined by the recommended methodology. The analysis strategy is complicated and difficult, and it contains limitations and compromises. It does not, however, require exotic analysis instrumentation, nor is it very expensive. Each of these facets of the methodology is discussed in this paper, and a status report is provided on development of a guidance document and a research project intended to produce improved methods. PMID- 21619232 TI - Quantitative enzyme-linked immunosorbent assay for determination of polychlorinated biphenyls in environmental soil and sediment samples. AB - An enzyme-linked immunosorbent assay (ELISA) for the quantitative determination of Aroclors 1242, 1248, 1254, and 1260 in soil and sediments was developed and its performance compared with that of gas chromatography (GC). The detection limits for Aroclors 1242 and 1248 in soil are 10.5 and 9 ng/g, respectively. The assay linear dynamic range is 50-1333 ng/g. Cross-reactivity of the assay with 37 structurally related potential cocontaminants in environmental soil samples was examined; none of the chlorinated anisoles, benzenes, or phenols exhibited >3% cross-reactivity, with <0.1% cross-reactivity being the norm. Soil spike recoveries of 107% and 104% were obtained for Aroclors 1242 and 1248, respectively, for a spike level of 5 mg/kg, with corresponding relative standard deviations of 14% and 17%. One hundred forty-eight environmental soil, sediment, and paper pulp samples, obtained from two EPA listed Superfund sites, were analyzed by ELISA and standard GC methods. Samples were extracted for ELISA analysis by shaking with methanol. Additional extractions of the same samples were performed either with supercritical carbon dioxide or by Soxhlet extraction with methanol. ELISA results for both the supercritical fluid and the Soxhlet extracts were in close agreement with the GC results, while the ELISA results for the methanol shake extracts were not. The data for the environmental samples demonstrated the capability of the ELISA to provide accurate results and reinforced the dependence of any detection method, including ELISA, on appropriate extraction procedures. PMID- 21619233 TI - Information theory approach to underdetermined simultaneous multicomponent analysis. AB - An algorithm for analysis of simultaneous multicomponent systems is proposed. This method is suitable for underdetermined systems, where the number of chemical components is larger than the number of measured data points. This method provides a rational solution to such underdetermined cases, with no need of any model assumptions. Such situations are faced, for example, in application of low cost sensors to environmental analysis, where complex mixtures must be handled. The proposed solution is based on information theory and provides a unique set of concentrations that is the most unbiased one. The algorithm maximizes the entropy of the solution set, which means that the final unjustified information is minimal. Detailed description of the mathematical procedure is provided. The method is exemplified for spectroscopic analysis and evaluated by extensive computer simulations. Several effects, such as the number of components and the number of measured points, the noise level, spectral characteristics, and the sampling design, are studied. The stability of the algorithm and the analytical performance in some cases are evaluated in respect to these factors. Satisfactory results are obtained in systems with realistic noise levels. PMID- 21619234 TI - Effect of capillary properties on the sensitivity enhancement in capillary/fiber optical sensors. AB - The sensitivity enhancement observed in fluorescence signals when a conventional fiber optical sensor is coupled with a quartz or glass capillary results from the partial reflection of the radiation at the sample/internal wall interface and from the internal reflection of the refracted portion within the capillary wall. Thus, the length and absorbing properties of the capillary as well as the nature of the surrounding medium affect the enhancement significantly. To interpret the dramatic changes in enhancement observed experimentally when the absorbing properties of the capillary were changed, a partial reflective waveguide model is reported. PMID- 21619235 TI - Direct electrochemistry for the imidazole complex of microperoxidase-11 in dimethyl sulfoxide solution at naked electrode substrates including glassy carbon, gold, and platinum. AB - The effect of the total water content on the persistence and rate of direct heterogeneous electron transfer between the imidazole complex of microperoxidase 11 (im-MP-11) and naked gold, platinum, and glassy carbon (GC) in dimethyl sulfoxide (DMSO) solutions containing 0.1 M tetra-n-butylammonium perchlorate was investigated using cyclic voltammetry. Electron transfer between im-MP-11 and Au, Pt, and GC has been found to be persistent for more than 1 h and at least quasi reversible [k(s)' = (8.7 +/- 0.1) * 10(-4) cm/s (Au), k(s)' = (7.2 +/- 1.3) * 10( 4) cm/s (Pt), and k(s)' = (5.7 +/- 1.0) * 10(-4) cm/s (GC)] in dimethyl sulfoxide containing an absolute water content between 0.1 and 1.8%. The heterogeneous electron transfer rate constant is independent of the total water content of the DMSO solution when between 0.1 and 1.8% water is present. PMID- 21619236 TI - Variable path length transmittance cell for ultraviolet, visible, and infrared spectroscopy and spectroelectrochemistry. AB - The design and characteristics of a transmittance cell for ultraviolet, visible, and infrared spectroscopy and spectroelectrochemistry are described. Through modification of a previously reported design, this cell employs threaded glass connectors as insertion ports for either quartz- or silicon-windowed tubes, thus permitting essentially continuous variation of the optical path length from ~0.050 to 200 mm. Though the initial fabrication requires skillful glassblowing, once constructed, the cell's simple design allows for rapid and reproducible disassembly/reassembly between experiments. The utility of the cell for a diversity of fluid samples is demonstrated through applications to water, aqueous ferricyanide, ferrocene in methylene chloride, and acetone vapor. PMID- 21619237 TI - Design and synthesis of sodium ion-selective ionophores based on 16-crown-5 derivatives for an ion-selective electrode. AB - To develop an ionophore that is highly selective for sodium for use in an ion selective electrode, we propose a model based on 16-crown-5 which has a cavity just the size of Na(+ )and has a "block" subunit to prevent complex formation with ions larger than Na(+). Based on this molecular model, eight kinds of 16 crown-5 derivatives have been synthesized, and their structural ion selectivity has been evaluated in detail. The 16-crown-5 derivatives having two bulky "block" subunits showed high Na(+) selectivity relative to K(+). In particular, the derivative with two decalino subunits (DD16C5) exhibited the highest Na(+) selectivity of all the ionophores examined. When a phosphate ester-type membrane plasticizer, tris(ethylhexyl) phosphate, was used as the membrane solvent for the ion-sensing membrane based on poly(vinyl chloride), the electrode using DD16C5 exhibited a Na(+) selectivity of over 1000 times relative to alkali metal and alkaline earth metal ions, including K(+), which is the most serious interferant. The evaluation of the relationship between the ionophore chemical structures and the ion-selective features contributes to the host-guest chemistry to give a highly selective ionophore for an alkali metal ion. PMID- 21619238 TI - Trace Analysis of Zn(II), Be(II), and Bi(III) by Enzyme-Catalyzed Chemiluminescence. AB - A novel technique for the trace analysis of metal ions Zn(II), Be(II), and Bi(III) in bulk solutions is discussed. This technique involves the generation of a chemiluminescence signal from alkaline phosphatase catalyzed hydrolysis of a phosphate derivative of 1,2-dioxetane. Zn(II) can be determined by two methods, reactivation of the alkaline phosphatase apoenzyme and inhibition of the native enzyme. Be(II) and Bi(III) can be determined quantitatively by inhibition of the native enzyme. Subppb to ppm level detection of Zn(II), Be(II), and Bi(III) has been achieved. Initial studies with mixed metals are also reported. The technique described is rapid and sensitive and can be readily applied to the microassay of heavy metal ions. PMID- 21619239 TI - Enhanced Serum Carbon Dioxide Measurements with a Silicone Rubber-Based Carbonate Ion-Selective Electrode and a High-pH Dilution Buffer. AB - A new silicone rubber matrix carbonate-selective membrane and a high-pH buffer diluent are used to enhance the performance of the electrode measurements for serum carbon dioxide. The proposed membrane employs one-component silicone rubber as the matrix and trifluoroacetyl-p-decylbenzene as the neutral ionophore. The optimized membrane formulation incorporates as high as 21.9 wt % plasticizer (e.g., bis(2-ethylhexyl) adipate). The highly plasticized silicone rubber membranes not only function equivalently, in terms of the carbonate response, to the conventional PVC matrix membranes, but they also exhibit substantially reduced interfering response toward salicylate. Furthermore, the silicone rubber membrane exhibits better adhesion to the solid surface than do PVC or PU membranes. The use of higher pH buffers (e.g., 2-amino-2-methyl-1-propanol (AMP) H(2)SO(4), pH 9.5-10.5) further enhances the selectivity of the carbonate electrode measurement system for total CO(2) species over other anions. It is shown that the combined use of the silicone rubber matrix membrane and the high pH AMP buffer provides a carbonate sensor system that is substantially less subject to interference from salicylate and chloride than is the conventional measurement system employing the PVC-based electrode with the lower pH (8.4-8.8) buffer diluent. PMID- 21619240 TI - A photochemical procedure for determining reaction quantum efficiencies in systems with multicomponent inner filter absorbances. AB - A kinetic procedure for measuring the photochemical quantum efficiencies of reactions in which several components have overlapping absorbances is fully described. This method has been found to be particularly suitable for photochemical determinations where inner filter effects are substantial, such as those typically observed in the ligand substitution and intermolecular C-H/Si-H bond activation mechanisms of metal complexes. The quantitative measurement of the Si-H bond activation photochemistry of (eta(5)-C(5)H(5))Rh(CO)(2) in triethylsilane solution is demonstrated in detail, although the procedure has widespread application in photochemistry. PMID- 21619241 TI - Aerodynamic Particle Sizing versus Light Scattering Intensity Measurement as Methods for Real-Time Particle Sizing Coupled with Time-of-Flight Mass Spectrometry. AB - Measurement of scattered light intensity and aerodynamic particle sizing are two methods that have recently been coupled with time-of-flight mass spectrometry for real-time determination of aerosol particle size and composition. An aerosol analysis technique recently developed in our laboratory, aerosol time-of-flight mass spectrometry, offers a unique experimental platform to evaluate both of these sizing techniques. This paper presents a comparison of results obtained with these two methods. PMID- 21619242 TI - Detection of nitric oxide and nitrogen dioxide with photoluminescent porous silicon. AB - The visible photoluminescence of porous Si is quenched by nitric oxide and nitrogen dioxide to detection limits of 1.4 * 10(-)(3) and 5.3 * 10(-)(5) Torr, respectively (corresponding to 2 ppm and 70 ppb). At analyte partial pressures in the low milliTorr range, the photoluminescence quenching is partially reversible; recovery from nitrogen oxide exposure occurs on a time scale of minutes. For both NO and NO(2), the reversible photoluminescence quenching response fits a Stern Volmer kinetic model. At higher partial pressures, quenching deviates from Stern Volmer kinetics and some permanent loss of photoluminescence intensity occurs due to oxidation of the porous Si surface. Photoluminescence from porous Si is not quenched by nitrous oxide or carbon dioxide and only slightly quenched by carbon monoxide and oxygen. PMID- 21619243 TI - Ion-exchange chromatography/electrospray mass spectrometry for the identification of organic and inorganic species in topiramate tablets. AB - An ion-exchange chromatograph/electrospray ionization mass spectrometer (IC/ESI MS) was used successfully to identify organic and inorganic species present in topiramate tablets. An ion suppressor is placed between the column and detectors to replace sodium ions in the mobile phase with hydrogen ions supplied by the suppressor. The ensuing combination of the hydrogen ions with the mobile phase hydroxide ions produces water and thus allows simultaneous ion detection by an ion conductivity detector and a mass spectrometer. Analytes, including lactate, glycolate, chloride, formate, sulfate, and oxalate, were unambiguously identified by matching the mass spectra and retention times with those of the authentic compounds. Due to its capability of detecting positive and negative as well as neutral species, ESI-MS provides valuable information which is not available with ion conductivity detection alone. Though the coupling of ion-exchange chromatography to mass spectrometry has been reported previously, this is the first demonstration of IC/ESI-MS for the identification of unknown species in real samples. Finally, with the use of deuterium/carbon-13 labeling and MS/MS techniques, we have confirmed that oxalic acid (HOOC-COOH) is formed from formic acid (HCOOH) at the electrospray interface in the presence of the electric field. This observation not only confirms the identity of an unknown peak, but it also provides new insight into chemistry that can take place during electrospray ionization. PMID- 21619244 TI - Efficient Ion Remeasurement Using Broadband Quadrupolar Excitation FTICR Mass Spectrometry. AB - Quadrupolar excitation is achieved over a wide mass range by using repetitive chirp excitation, filtered noise excitation, and high-amplitude, single-frequency excitation for matrix-assisted laser desorption FTICR mass spectrometry. These methods efficiently axialize ions over a wide range of masses in a 4.7 T FTICR spectrometer. Remeasurement efficiencies are >99.5% for broadband repetitive chirp excitation, 99% with filtered white noise, and 99.4% with high-amplitude, single-frequency broad-band quadrupolar excitation. Results indicate that z-axis ejection of ions during detection is the primary mechanism for ion loss during remeasurement experiments. Capacitive coupling of the excitation signal to the trapping plates of the open-ended cylindrical analyzer cell is required for high remeasurement efficiency. PMID- 21619245 TI - Purification, recovery, and laser-driven fluorination of silicon from dissolved and particulate silica for the measurement of natural stable isotope abundances. AB - A procedure for the purification, recovery, and determination of isotopic abundances of silicon from biogenic and lithogenic particulate matter and dissolved silicic acid is reported. Purification involves the reaction of acid molybdate with dissolved silicon in natural waters or that produced by the dissolution of particulate silica by hydrofluoric acid. The resulting silicomolybdic acid is then quantitatively precipitated by reaction with triethylamine hydrochloride. The silicon is recovered as silicon dioxide through stepwise combustion of the dried precipitate. Fluorination of the product for isotopic analysis is accomplished by laser heating under pure fluorine generated by the decomposition of a fluorine-based salt. The resulting silicon tetrafluoride is separated from hydrogen fluoride and other fluorination byproducts cryogenically using a variable-temperature cold trap. Yields for silicon recovery are 99.9% for precipitation and greater than 95% for the purification/fluorination procedure. Reproducibility of the isotopic composition for pure quartz granules processed through the procedure is +/-0.10/00 for delta(30)Si. PMID- 21619246 TI - Grafting Poly(ethylene glycol) Epoxide to Amino-Derivatized Quartz: Effect of Temperature and pH on Grafting Density. AB - Microparticle capillary electrophoresis was used to characterize the surface of quartz capillaries grafted with the glycidyl ether of poly(ethylene glycol) (E PEG). Site dissociation modeling of capillary electrokinetic behavior provided estimates of surface group pK and density, plus the distance (d) from the surface to the hydrodynamic plane of shear. Native quartz appeared to possess silanol groups of pK 3.6 and 6.9 whose surface densities varied with quartz treatment. Aminopropylsilane derivatization of quartz silanol groups in toluene yielded a coating which was stable (>6 h) at pH 10.3 and 60 degrees C. Aqueous grafting of E-PEG to this surface was relatively independent of pH (7.3-10.3) and reaction time (6-24 h) but was significantly influenced by reaction temperature (25-95 degrees C) and salt composition. PEG-grafted capillaries exhibited greatly reduced electroosmosis from pH 2 to 11. Significant grafting could be obtained under mild conditions (6 h, 35 degrees C, 0.4 M K(2)SO(4), pH 6.9). These results suggest that PEG chains increasingly extend normal to a surface as their grafting density increases, and that PEG conformation influences grafting density. The methods described should aid the use of PEG-coated surfaces in a variety of applications. PMID- 21619247 TI - Selective in vivo and in vitro sampling of proteins using miniature ultrafiltration sampling probes. AB - The most widely used in vivo sampling technique, microdialysis sampling, provides important data on the extracellular concentration of low molecular mass (<1000 5000 Da) species. However, biological macromolecules of much greater mass (>20-90 kDa) have key in vivo roles as chemical messengers or are currently under consideration as biopharmaceuticals. Microdialysis, which utilizes a sampling process based upon analyte diffusion, is largely ineffective at monitoring the local, transient extracellular concentrations of important macromolecular species. Ultrafiltration sampling is an in vivo sampling technique utilizing a convective, rather than diffusive, sampling mechanism. This paper demonstrates the effective recovery (>90%) of model proteins with molecular weights up to 68 000 from in vivo and in vitro sites through the use of miniature ultrafiltration sampling probes. Selectivity in the sampling process can be achieved through alteration of the membrane structure. In vivo ultrafiltration sampling in conjunction with slab gel electrophoresis and silver staining detects three recovered proteins (MW 9100-26 800) present in the extracellular space of a series of rats. PMID- 21619248 TI - Application of rhodamine 800 for reversed phase liquid chromatographic detection using visible diode laser-induced fluorescence. AB - This study reports the application of rhodamine 800, a far-red dye, suitable for excitation using visible diode laser-induced fluorescence (VDLIF) detection. A reagent synthesized from rhodamine 800 was evaluated as a precolumn reagent for derivatization with amino-containing analytes. The derivative of this reagent with primary amine analytes showed a loss of fluorescence. Rhodamine 800 was then applied as a mobile phase additive in the indirect mode for quantitation of valproic acid in plasma using reversed phase HPLC in combination with VDLIF detection. A visible diode laser (output power 8.50 mW) temperature-tuned to oscillate at 674.70 nm was used as a light source for a laboratory constructed HPLC fluorescence detector. A liquid/liquid extraction procedure was applied to human blank plasma. The selectivity of this method was validated by demonstration of a lack of interfering peaks in extracts of plasma (n = 3 sources). A calibration curve for valproic acid between 40 and 200 MUg/mL was shown to be linear (r = 0.9932). The recoveries of valproic acid at concentrations of 50 and 100 MUg/mL were evaluated and determined to be 73 and 72%, respectively. The precision and accuracy (n = 5) of the assay was within 7.0% RSD and 8.0% difference from the spiked concentration, respectively. The limits of detection (S/N = 3) for extracted and unextracted valproic acid were 15.0 and 11.54 MUg/mL, respectively. The theoretical (C(lim)) and practical (C(det)) limits of detection in the detector flow cell for unextracted valproic acid at a S/N = 1 were found to be within 15%. PMID- 21619249 TI - Molecularly imprinted polymer beads: suspension polymerization using a liquid perfluorocarbon as the dispersing phase. AB - A suspension polymerization technique suitable for molecular imprinting is described, based on the use of a liquid perfluorocarbon as the dispersing phase. This dispersant does not interfere with the interactions between functional monomers and print molecules required for the recognition process during molecular imprinting. The method produces polymer beads, with almost quantitative yield, which can be used after only a simple washing step. An acrylate polymer with perfluorocarbon and poly(oxyethylene) ester groups was used to stabilize an emulsion of functional monomer, cross-linker, print molecule, initiator, and porogenic solvent in perfluoro(methylcyclohexane). Initiation of polymerization by UV irradiation resulted in polymer beads. The average bead size could be controlled between about 50 and 5 MUm by varying the amount of stabilizing polymer. SEM of the beads indicated spherical particles with morphology typical of beads made by suspension polymerization. The technique was applicable to a range of conditions typically used for molecular imprinting. A detailed chromatographic study of the polymer beads confirmed that alpha values and resolution factors were similar to those achieved with traditional ground and sieved imprinted polymers. Small (5 MUm) beaded packings gave low back pressure and rapid diffusion, giving good separation even at high flow rates. PMID- 21619250 TI - Determination of the enantiomeric composition of chiral amines based on the quenching of the fluorescence of a chiral calixarene. AB - The synthesis of three chiral calixarene derivatives is described. One of these, an (S)-di-2-naphthylprolinol tetramer, is shown to exhibit significant ability to discriminate between enantiomers of 1-phenylethylamine (PEA) and norephedrine on the basis of the quenching of the (S)-di-2-naphthylprolinol fluorescence emission in chloroform. The chiral discrimination appears to arise from preorganization of the four (S)-di-2-naphthylprolinol substituents on the calixarene, which define a three-dimensional chiral space. The ability to measure the enantiomeric composition of PEA and norephedrine to within an error of 4.1% and 2.6%, respectively, on the basis of a single fluorescence measurement is demonstrated. PMID- 21619251 TI - Determination of (234)th in marine samples by liquid scintillation spectrometry. AB - A liquid scintillation spectrometry method for the determination of (234)Th in seawater with (230)Th as the yield tracer has been developed and validated. (234)Th is separated from the dissolved phase by an Fe(OH)(3) precipitation and is then purified using ion exchange chromatography. The counting source is prepared by taking the sample to dryness in a vial, redissolving in acid, and mixing with a scintillation cocktail. The instrument employed has a relatively low background (11 cpm) and the ability to separate alpha from beta activity on the basis of pulse shapes. The (234)Th + (234m)Pa counting efficiency is 50% over the counting window employed. The limit of detection, using the above parameters, a 20 L sample, and a 400 min count is found to be 0.04 dpm L(-)(1). It was also demonstrated that less advanced instruments, without alpha/beta separation, can also be used effectively. PMID- 21619252 TI - Cathodic Stripping Voltammetry of Selenium(IV) at a Silver Disk Electrode. AB - A new, simple, and reproducible method is described for the determination of selenium(IV) based on differential pulse cathodic stripping voltammetry. The optimized experimental conditions are as follows: selenium(IV) ions in an acidic medium (0.06 M HCl-0.07 M HNO(3)) are electrodeposited on a rotating silver disk electrode as silver selenide at -0.4 V vs SCE for 30 min; the deposit is then cathodically stripped in another solution (2 M NaOH) at a scan rate of 50 mV s( 1) to -1.2 V vs SCE. The cathodic stripping results in only a single well-defined peak at about -0.95 V vs SCE. The calibration (peak height vs selenium concentration) graph is linear up to at least 40 ng mL(-1) of selenium(IV) and passes through the origin, with a relative standard deviation of 2.7% for 20 ng mL(-1) (n = 5). The detection limit (3sigma) is 0.20 ng mL(-1). The possible interferences have been evaluated. Dissolved oxygen does not affect the peak height of selenium. The electrode can be used repeatedly at least 20 times with excellent reproducibility without further polishing. The proposed method is an improvement over the existing cathodic stripping techniques. PMID- 21619253 TI - Density and solubility of nafion: recast, annealed, and commercial films. AB - The density of Nafion membranes wetted in water and acetonitrile and the solubility of Nafion in a 50:50 ethanol/water mixture are reported. Commercially available membranes, solution recast membranes, and recast membranes thermally processed for varying lengths of time were examined. Unheated recast membranes were found to be ~20% less dense in water than both commercial membranes and recast membranes heated at 140 degrees C for 40, 50, and 60 min. For heating times of >=40 min, the density of recast membranes in water was invariant and equal to the density of commercial films. Similarly, in acetonitrile, commercial membranes and films heated for 1 h had the same density; unheated recast films were ~25% less dense. Nafion density in acetonitrile was ~15% less than that in water. Acid pretreatment reduced Nafion film density by ~15%. Unheated recast films were 22-100% soluble in a sonicated ethanol/water mixture. Commercial membranes and recast films heated for as little as 10 min were insoluble under the same conditions. PMID- 21619254 TI - Selective electrochemical detection using a split disk array electrode in a thin layer radial flow system. AB - An eight-sector array (split disk) electrode was designed for a low flow rate (<100 MUL/min) amperometric detector. This electrode was fabricated photolithographically for dimensional accuracy and reproducibility. This array of a pie-shaped electrode was combined with a thin-layer radial flow cell, and a conversion efficiency of 94% was achieved at the lowest flow rate tested (0.01 mL/min). Each electrode worked free from the effects of electrochemical reactions of the other electrodes. A coulometric hydrodynamic voltammogram of reversible redox species obtained using this system exhibited a Nernstian curve. These properties enabled this electrochemical detector to be used for determining the ratio of two redox species (redox potential difference ~ 100 mV) with small injection volume (5 MUL). PMID- 21619255 TI - Continuous-Flow System for On-Line Water Monitoring Using Back-Side Contact ISFET Based Sensors. AB - A multiparametric continuous-flow system for on-line monitoring of water based on ISFET sensors is described. The ISFETs used have silicon nitride as gate material, and the electrical contacts are placed on the back side of the chip. This is a technological improvement that allows for a more compact ISFET packaging and greatly increases the lifetime of the sensor compared with planar type ISFETs, since the electrical parts are separated from the chemical environment. A special probe has been designed in order to encapsulate and apply these ISFETs into the flow system. Further, a reference electrode based on standard Ag/AgCl technology has been constructed according to the ISFET probe design in order to integrate both sensors in the same flow-through cell. These probes can be easily replaced in the flow system and are made of cheap and easily mechanized materials. Using these flow-through sensors, a continuous-flow system for the determination of pH, NH(4)(+), Ca(2+), and NO(3)(-) in waters has been designed. The system configuration is based on a modular design (one setup for each parameter and a common sampling channel), which allows simple manipulation and maintenance as well as a good flexibility for different analytical requirements. A study of the system characteristics was performed by following the specifications for water monitoring. Under the conditions established for the flow system, a sampling rate of 20 h(-)(1) was obtained for each parameter, and long-term stabilities of at least 3 weeks of daily work for ISFET sensors and 5 months for the reference electrode have been achieved. The response performances obtained show the feasibility of the BSC ISFET probe use in continuous-flow monitoring. PMID- 21619256 TI - Flow injection based renewable electrochemical sensor system. AB - A novel class of electrochemical sensors is proposed utilizing electrically conducting beads to form a disposable electrode as well as nonconducting beads to form renewable layers of immobilized enzymes. The concept, aimed to prevent fouling, is tested on an amperometric sensor coupled to nonconducting beads with different immobilized oxidases: galactose, lactate, alcohol, or glucose oxidase, the latter two being used to determine alcohol and gluocse, respectively, in samples of beer and wine. Glucose oxidase was also immobilized on conducting glassy carbon particles to explore the performance of a biosensor where both enzyme and electrode can be automatically renewed in less than 1 min. The results confirm that the concept of a flow injection renewable electrochemical sensor (FI RES) is practical. It provides a novel approach to biosensing, to comparing enzyme activity, to studying enzyme immobilization on different supports, and to voltammetry in general. PMID- 21619257 TI - Fast-scan voltammetry of cyclic nitroxide free radicals. AB - Fast-scan cyclic voltammetry at carbon fiber microelectrodes is used to detect the cyclic nitroxide 2,2,6,6-tetramethylpiperidinyl-1-oxy free radical (TEMPO) and three analogs. The electrochemical behavior of the TEMPO analogs at unmodified carbon fiber electrodes is found to differ greatly from their behavior at glassy carbon electrodes. After the electrode is coated with the polymer Nafion, the electrodes exhibit increased sensitivity to TEMPO and 4-amino-TEMPO. Voltammograms of the nitroxides at Nafion-coated electrodes indicate that the oxidized form (oxoammonium ion) and the free radical form have greatly different mobilities through the polymeric coating. Response times to changes in nitroxide concentration vary from subsecond at bare electrodes (all four analogs) and 4 hydroxy-TEMPO at modified electrodes to 1-3 s for TEMPO and 4-amino-TEMPO at modified electrodes. The detection limit for 4-amino-TEMPO is 50 MUM at an unmodified electrode and 5 MUM at a Nafion-coated carbon fiber electrode. The sensitivity of the Nafion-modified electrode to TEMPO, 4-hydroxy-TEMPO, and 4 amino-TEMPO can be improved by choosing a resting potential at which the oxoammonium ion form of the nitroxide is preconcentrated into the Nafion film. Using fast-scan cyclic voltammetry and the modified carbon fiber electrodes, the reaction of two nitroxide free radicals with ascorbate can be monitored. This work shows that fast-scan voltammetry at microelectrodes is a sensitive method that can be used to follow reactions of cyclic nitroxide free radicals in solution. PMID- 21619258 TI - Electrostatic spraying: a novel technique for preparation of polymer coatings on electrodes. AB - A liquid flow emerging from a tip or a thin tube under the influence of a strong electric field will, due to charging of the dielectric liquid, break up into small droplets. Thus, if a polymer material is dissolved in the liquid, this electrodeposition technique can be utilized for producing polymer coatings on electrodes. The method was applied for in situ formation of ultrathin (~3000 A) cellulose acetate (CA) phase inversion membranes on glassy carbon electrodes. The purpose of the membrane was to protect the electrode surface from fouling by macromolecular species. The spraying liquid consisted of CA, acetone, and aqueous magnesium perchlorate as pore former, and the spraying voltage was 14 kV. Profilometric measurements showed that the thickness of the spray-cast membranes was much more uniform than that of similar membranes formed by solvent casting. By using cadmium and lead as test analytes and differential pulse anodic stripping voltammetry as detection method, it was found that the membranes prepared by spray casting offered better protection against interference from poly(ethylene glycol) (PEG) 6000 than those prepared by solvent casting. Also, the interference from PEG 2000 was significantly reduced. Experimental details of the electrostatic spraying technique are given. PMID- 21619259 TI - Simultaneous mixture analysis using a dynamic microbial sensor combined with chemometrics. AB - A biosensor consisting of immobilized microbial cells and an oxygen electrode was used in a flow-through system as a microbial sensor flow injection analyzer (FIA). For different organic analytes, the metabolism of vital cells provides individual time-resolved responses with distinct time-dependent amplitudes. Chemometrical data analysis revealed that the individual responses are additive and depend linearly on single analyte concentrations. Based on these observations, simultaneous multicomponent analysis of organic mixtures was carried out in the FIA's time domain with analytical errors of less than 10%. For mixture analysis and monitoring in processes like enzymatic conversions, the described microbial sensor FIA ("dynamic microbial sensor") offers an alternative to expensive analytical equipment. PMID- 21619260 TI - Elimination of uninformative variables for multivariate calibration. AB - A new method for the elimination of uninformative variables in multivariate data sets is proposed. To achieve this, artificial (noise) variables are added and a closed form of the PLS or PCR model is obtained for the data set containing the experimental and the artificial variables. The experimental variables that do not have more importance than the artificial variables, as judged from a criterion based on the b coefficients, are eliminated. The performance of the method is evaluated on simulated data. Practical aspects are discussed on experimentally obtained near-IR data sets. It is concluded that the elimination of uninformative variables can improve predictive ability. PMID- 21619261 TI - Development, validation, and application of a method for quantification of methylmercury in biological marine materials using gas chromatography atomic emission detection. AB - A gas chromatographic method was developed for the quantification of alkylmercury species using microwave-induced plasma atomic emission detection (GC-AED). The column conditioning and analyte derivatization required for previous methods were found to be unnecessary for stable, accurate, and sensitive element-specific detection using GC-AED. Chromatographic and detection parameters such as stationary phase type, stationary phase film thickness, GC column dimensions, helium mobile phase column head pressure, detector makeup gas flow rate, and detector reagent gas type and flow rate were found to significantly affect analyte response. The detection limit for the optimized GC-AED conditions was 0.8 pg (0.1 pg/s) of methylmercury chloride (as mercury). A solid-liquid extraction procedure with preparative gel permeation chromatography cleanup and GC-AED analysis was used to quantify methylmercury in a variety of complex matrix marine materials. The methylmercury quantification method was validated with four marine certified reference materials (CRMs). The method was then applied to 13 standard reference materials, CRMs, and control materials for which no certified reference values for methylmercury have been determined. Four National Institute of Standards and Technology Standard Reference Materials and one control material, which were analyzed using the GC-AED method, were also analyzed by two other laboratories using independent methods to further validate the method. PMID- 21619262 TI - Determination of acaricides in honey by liquid chromatography with ordinary, narrow-bore, and microbore columns. AB - Narrow-bore and microbore columns packed with octadecylsilane were used to compare their sensitivity and efficiency in the separation of coumaphos, fluvalinate, bromopropylate, and 4,4'-dibromobenzophenone from honey with those of ordinary columns. The best sensitivity for acaricides was accomplished by using a 150 mm * 0.32 mm i.d., 5 MUm Spherisorb ODS-2 capillary column, methanol water (90:10 v/v) as the mobile phase, and 5 MUL/min as the flow rate. Detection limits for individual acaricides using a UV detection range from 0.40 to 0.74 MUg/kg of honey were comparable to those obtained by gas chromatography using an electron capture detector. All acaricides were separated in <12 min. The coefficients of variations on real samples were <6.2%. PMID- 21619264 TI - Chromatography of mechanically interlocked molecular compounds. AB - Catenanes and rotaxanes are molecules composed of mechanically interlocked components which are not linked to each other by covalent bonds. These molecular assemblies behave as discrete molecules with defined properties significantly different from those of the parent "free" components. High-performance liquid chromatography has been employed successfully to characterize some tetracationic catenanes and rotaxanes incorporating either cyclobis(paraquat-p-phenylene) or cyclobis(paraquat-4,4'-biphenylene) as the charged components and either hydroquinone-containing macrocycles or dumbbell-shaped entities as the neutral components. In each case, significant differences in the retention times of the mechanically interlocked molecular compounds, in comparison with those of their components as their "free" forms, were observed. PMID- 21619263 TI - Theory and Practice of Rapid Flow-Through Analysis Based on Optode Detection and Its Application to pH Measurement as a Model Case. AB - Rapid flow-through analysis (RFA) based on optode (optical chemical sensor) detection is proposed, and its performance is discussed using the RFA system equipped with a pH-sensitive optode as a model case. To demonstrate and understand the usefulness of the RFA system, long-lifetime pH-sensitive optodes were prepared using a hydrophilic poly(hydroxyethyl methacrylate) (poly-HEMA) membrane covalently immobilized with a dye containing an amino group such as Congo Red or Nile Blue. The response equation for the optode with the RFA system was proposed, and satisfactory estimation for rapid pH analysis was obtained. The proposed RFA can stand as an advanced method for conventional flow injection analysis. PMID- 21619265 TI - Xylene Isomer Mass Spectral Identification through Metal Ion Chemistry in an FTICR. AB - The ortho, meta, and para isomers of xylene are identified in a Fourier transform mass spectrometer by reactions with V(+) and VO(+). Each isomer reacts in a unique way with a mixture of the two ions, which enables isomer discrimination. The method is fast and does not require the use of internal standards. PMID- 21619266 TI - Broad-band fourier transform quadrupole ion trap mass spectrometry. AB - Broad-band nondestructive ion detection is achieved in a quadrupole ion trap mass spectrometer by impulsive excitation of a collection of trapped ions of different masses and recording of ion image currents induced on a small detector electrode embedded in but isolated from the adjacent end cap electrode. The image currents are directly measured using a simple differential preamplifier, filter, and amplifier combination and then Fourier analyzed to obtain broad-band frequency domain spectra characteristic of the sample ions. The use of the detector electrode provides a significant reduction in capacitive coupling with the ring electrode. This minimizes coupling of the rf drive signal, which can saturate the front-end stage of the detection circuit and prevent measurement of the relatively weaker ion image currents. Although impulsive excitation is preferred due to its broad-band characteristics and simplicity of use, results are also given for narrow-band ac and broad-band SWIFT (stored wave-form inverse Fourier transform) excitation. Data using argon, acetophenone, and n-butylbenzene show that a resolution of better than 1000 is obtained with a detection bandwidth of 400 kHz. An advantage of nondestructive ion detection is the ability to measure a single-ion population multiple times. This is demonstrated using argon as the sample gas with an average remeasurement efficiency of >90%. Tandem mass spectrometry experiments using a population of acetophenone ions are also shown. PMID- 21619267 TI - Graphite/Liquid mixed matrices for laser desorption/ionization mass spectrometry. AB - A combination of liquid matrix and graphite particulates (2 MUm) has been proposed as a method suitable for the laser desorption/ionization mass spectrometry of peptides and proteins (Sunner, J.; et al. Anal. Chem. 1995, 67, 4335). Here we demonstrate the potential of this approach as a straightforward, and very general, method of achieving the ultraviolet laser desorption/ionization of a broad range of intermediate weight analytes. The desorption/ionization mechanism, the influence of preparative procedures, and the breadth of application of this methodology have been investigated. A simple and robust preparative procedure is presented for the analysis of proteins, oligosaccharides, and synthetic polymers. Detection sensitivities are in the femtomole region for lower molecular weight peptides and oligosaccharides. The graphite acts as an energy transfer medium by absorbing the UV radiation, leading to thermal desorption of the liquid matrix and analyte. The liquid matrix was observed to fulfill several important roles. In the case of peptides and proteins, which preferentially form protonated molecular ions, it acts as a protonating agent. It also enhances the signal intensities of cationized species (e.g., polysaccharides and polar polymers) by assisting their desorption. An excess of liquid matrix serves to cool the analyte during the desorption step and minimize decomposition. The presence of liquid matrices increases the sample lifetime at a particular desorption spot, minimizing the time-consuming search for "hot spots". The addition of cationizing salts has been shown to improve the quality of mass spectra obtained for polar polymers and extend the range of materials that can be investigated to include apolar synthetic polymers. PMID- 21619268 TI - Electrochemical and XPS Study of the Nickel-Titanium Electrode Surface. AB - A Ni-Ti alloy with a 50:50 atomic composition has shown exceptional properties as a fixed potential LCEC detector for carbohydrates and related substances. It exhibited excellent sensitivity and superior long-term stability compared to pure Ni. A study was therefore undertaken by means of cyclic voltammetry (CV), scanning electron microscopy (SEM), and X-ray photoelectron spectroscopy (XPS) to understand the role of Ti and the respective surface oxides of Ni and Ti in the catalytic stability of the detector. CV results showed that Ti is initially oxidized, most likely to TiO(2) in 0.1 M NaOH solution. The oxidation of Ni to nickel(II) oxide also occurs at potentials close to that of Ti. At higher potentials in the range of +0.4 to +0.5 V vs Ag/AgCl reference, nickel(II) oxide undergoes further oxidation to the Ni(III) oxidation state. This state is responsible for the catalysis of carbohydrates, amino acids and other biosubstances. When Ni-Ti and Ni are repetitively CV cycled in the potential range of 0.0 to +0.6 V, a second wave appears at more negative potentials during the reverse cathodic scan for Ni but not for Ni-Ti. SEM images of these two electrodes in the oxidized form show the Ni-Ti surface remains smoother in appearance. This smoothness is consistent with the fact that the thickness of the surface "oxide" layer increases less rapidly, as Ni-Ti is repetitively CV cycled, compared to pure Ni. XPS results for the nature of the surface oxides are consistent with oxidized Ti as TiO(2), Ni(II) predominantly as Ni(OH)(2), and Ni(III) possibly as NiOOH. Possible reasons for Ti stabilizing the Ni-Ti alloy as a LCEC detector are discussed. PMID- 21619269 TI - Fabrication, characterization, and potential application of carbon fiber cone nanometer-size electrodes. AB - A novel method has been developed for the fabrication of carbon fiber cone nanometer-size ultramicroelectrodes (nanoelectrodes) with overall tip dimensions as small as 50 nm in diameter. In this method, carbon fibers were initially etched by an argon ion beam thinner. Afterward, a single etched carbon fiber was inserted into a glass capillary, which was then sealed by heating the glass/fiber interface in a vacuum; thus, no epoxy resin is involved. The success rate of our fabrication route for the electrodes with overall tip diameters of up to 500 nm was about 80%; for those with tip diameters of up to 100 nm, it was about 50%. The fabricated carbon fiber cone nanoelectrodes (CFCNEs) were inspected by optical and scanning electron microscopy. Their electrochemical behavior was examined by cyclic and linear sweep voltammetric measurements of ferricyanide and ferrocene ions in aqueous and nonaqueous media. The potential analytical applicability of the CFCNEs was tested by differential pulse voltammetric measurements of two well-known neurotransmitters, dopamine (DA) and 5 hydroxytryptamine (5-HT), and the results achieved were highly satisfactory. The calibration plots obtained were linear over the ranges from 5.0 * 10(-7) to 1.0 * 10(-4) and from 2.0 * 10(-6) to 1.0 * 10(-4) mol/L, with limits of detection of 1.0 * 10(-7) and 5.0 * 10(-7) mol/L for DA and 5-HT, respectively. Some advantages and improvements of the proposed CFCNE fabrication method, especially with respect to smoothness of the fiber (electrode) surface, strength, and control over the fiber tip dimensions, are also discussed. PMID- 21619270 TI - Detection of hydrazine, methylhydrazine, and isoniazid by capillary electrophoresis with a palladium-modified microdisk array electrode. AB - A palladium particle-modified carbon fiber microdisk array electrode was designed and employed in capillary electrophoresis for the simultaneous detection of hydrazine, methylhydrazine, and isoniazid. The Pd-modified microdisk electrode had high catalytic ability for hydrazines and exhibited good reproducibility and stability. The response for hydrazine was linear over 3 orders of magnitude with a correlation coefficient of 0.993. The detection limits for hydrazine, methylhydrazine, and isoniazid were 1.2, 2.1, and 6.2 pg, respectively. PMID- 21619271 TI - Identification of chemical interferences in aldehyde and ketone determination using dual-wavelength detection. AB - A method for the rapid and convenient identification of chemical interferences in the determination of aldehydes and ketones in air samples using the 2,4 dinitrophenylhydrazine (DNPH) method is described. The ratio of absorptions at 360 and 300 nm is characteristic for groups of related aldehydes and ketones as well as for the main interferents. It has been determined by UV/visible spectroscopy of pure standard compounds and confirmed by HPLC analysis with UV/visible detection of complex hydrazone mixtures. The application of this method has led to the identification of 2,4-dinitrochlorobenzene as another interfering compound after sampling of air with high nitrogen dioxide content when using hydrochloric acid as catalyst. The possibilities and limitations of the dual-wavelength detection with the DNPH method are discussed for mixtures of standards and real samples from car exhaust. PMID- 21619272 TI - Laser two-photon ionization of pyrene on contaminated soils. AB - We report a first attempt to use the laser multiphoton ionization method for analysis of trace aromatic compounds on the surface of environmental (soils) and artificial (silica) samples. The measurement setup is composed of a N(2) pulsed laser and a fast conductivity detection system. The technique has been tested for detection of pyrene deposited on moist fine-powdered samples. The observed photoionization signals have indicated a gradual increase in the photoionization current and charge as a function of increasing concentration of pyrene/hexane solutions used for sample contamination. Contaminants have been analyzed in several (organic and inorganic) environmental samples, and a method to compensate for matrix effects is suggested. A contamination model is assumed and applied in order to renormalize all signals and provide an useful calibration plot. This calibration plot provides an upper estimate of pyrene LOD as 35 ng/g. PMID- 21619273 TI - Microformat imaging ELISA for pesticide determination. AB - A flat-well microformat competitive enzyme-linked immunosorbent chemiluminescent assay for the detection of the pesticide 2,4-dichlorophenoxyacetic acid (2,4-D) is described. Thick-film technology was used to pattern a hydrophobic layer 100 MUm thick onto glass microscope slides to form an array of 2 * 2 mm(2) squares. These flat wells were able to hold 2 MUL of reagents, corresponding to a height of ~500 MUm, with minimal contamination risk. The hydrophobic ink used to pattern the surfaces allowed significantly larger volumes of samples to be applied when compared with surfaces patterned with nonhydrophobic inks. This reduced evaporation effects and permitted greater pipetting accuracy, thereby improving assay reproducibility. A competitive immunoassay was developed based on the ability of free 2,4-D hapten to inhibit binding of anti-2,4-D monoclonal antibodies to 2,4-D-bovine serum albumin conjugate adsorbed onto the glass support. The support was subsequently incubated with alkaline phosphatase (AP) labeled anti-mouse IgG. The amount of AP conjugate bound was determined by quantitating the chemiluminescent emission produced from the enzymatic breakdown of CSPD substrate by AP using a cooled CCD camera. The detection limit of the single-sample microformat assay was 2.7 * 10(-)(11) M, or 6 pg of 2,4-D. The linear ranges of the single-sample and multisample assays were 4.5 * 10(-)(8)-4.5 * 10(-)(11) and 4.5 * 10(-)(7)-1.66 * 10(-)(10) M, respectively. In comparison, the detection limits of a tube-based chemiluminescent assay using standard luminometer and of a colorimetric ELISA were 45 * 10(-)(11) and 9.9 * 10(-)(8) M, respectively. The ability to scale the thick-film-based microformat assay makes it an ideal candidate for the development of affinity arrays and high-throughput assay formats. Prospects for further improvements of this imaging ELISA strategy will be discussed. PMID- 21619274 TI - Biosensor for the nonspecific determination of ionic surfactants. AB - We report the analytical figures of merit for the first biosensor for ionic surfactant quantification. The biosensor consists of a silanized silica optical fiber, onto which acrylodan-labeled bovine serum albumin (BSA-Ac) is immobilized. Recent work from our laboratory on native BSA-Ac (Lundgren, J. S.; Bright, F. V. J. Phys. Chem. 1996, 100, 8580-8586) has shown that ionic surfactants dehydrate the local environment surrounding the lone acrylodan residue, open up the pocket hosting the acrylodan reporter group, and dramatically increase the segmental mobility of domain I in BSA. In the current work, we use BSA-Ac as an actual biorecognition element for surfactant detection and quantification. We also compare several BSA-Ac immobilization strategies and determine the analytical figures of merit for the BSA-Ac-based biosensor to a prototypical analyte, cetyltrimethylammonium bromide (CTAB). The biosensor linear dynamic range extends from 5 to 60 MUM, and the t(90) response time (90% of the total response) is less than 30 s. Biosensor response precision (relative standard deviation) during 34 sensing cycles is 2.5%. On the down side, biosensor performance decreases 38% after 25 days of storage; however, this drift can be compensated. This work also demonstrates the utility of BSA-Ac as a model biorecognition element-reporter group system for grading the suitability of different surface immobilization strategies. PMID- 21619275 TI - Acceptance Criteria for Ultratrace HPLC-Tandem Mass Spectrometry: Quantitative and Qualitative Determination of Sulfonylurea Herbicides in Soil. AB - Eight commonly used sulfonylureas (SUs: nicosulfuron, thifensulfuron methyl, metsulfuron methyl, sulfometuron methyl, chlorsulfuron, bensulfuron methyl, tribenuron methyl, and chlorimuron methyl) and deuterium-labeled nicosulfuron (nicosulfuron-d(6)), used as an internal standard, were isolated from soil by solvent extraction and identified under quantitative and qualitative ion spray LC/MS/MS conditions using the selected reaction monitoring (SRM) mode of acquisition. The lower level of quantitation for these SUs in soil was determined at the 0.05 ppb level using a TurboIonSpray adapted LC/MS interface without a precolumn split and optimizing MS/MS tuning conditions for individual SUs. The eight SUs were qualitatively identified and quantitatively determined in soil. The standard curve for each SU was linear from 0.05 to 10 ppb. This SRM LC/MS method demonstrates high sensitivity and high specificity for these SUs in soil and shows at least a 400-fold improvement in sensitivity over previous reports. Acceptance criteria for forensically valid data are suggested for qualitative SRM LC/MS experiments. These include HPLC retention time reproducibility (+/-2%), at least two and preferably three precursor-product ions selected, and relative abundance criteria for selected ions (+/-20% absolute). PMID- 21619276 TI - Effectiveness of Purge-and-Trap for Measurement of Volatile Organic Compounds in Aged Soils. AB - The U.S. EPA-recommended method for measurement of trace levels of volatile organic compounds (VOCs) in soil, purge-and-trap, measures the readily desorbable organic contaminants from soil pore spaces and external soil surfaces. It does not, however, measure contamination that has diffused into internal micropores of soil matrix. Thus, the purge-and-trap method measures only a small fraction of total soil contaminants, especially in long-contaminated soils, where ~90-99% of contamination may be in the interior of the soil matrix. We compared three methods for determination of VOCs in aged field samples: purge-and-trap, methanol immersion, and hot solvent extraction. Hot solvent extraction proved to be much more effective than the U.S. EPA-approved purge-and-trap technique. For three long-contaminated soils containing such VOCs as trichloroethene, benzene, toluene, chloroform, methylene chloride, and cis-1,1-dichloroethylene, recovery from purge-and-trap ranged between 1.5 and 41.3% that of hot solvent extraction. Our data show that purge-and-trap may not be the best methodology for measuring soil VOCs concentrations, particularly in aged soils. It is clear from this and previous studies that the best overall choice for soil VOCs measurements is hot solvent extraction. These results also indicate the inefficiency of purge-and trap as a method for evaluating vapor extraction remediation technology. Our results suggest that the EPA should review the use of the purge-and-trap method for measuring VOCs concentrations in soils. PMID- 21619277 TI - Liposome behavior in capillary electrophoresis. AB - The behavior of liposomes in capillary electrophoresis is studied for the purpose of developing a potential method for characterizing liposomes prepared for use in industrial and analytical applications. This study characterizes the electrophoretic behavior of liposomes under various conditions to provide information about electrophoretic mobility and liposome-capillary surface interactions. The results of this method are compared with the results obtained using traditional laser light-scattering methods to obtain size information about liposome preparations. Additionally, reactions of liposomes and the surfactant n octyl-beta-d-glucopyranoside are performed off-line in bulk solution experiments and on-line in the capillary. Automated delivery of lysis agents by multiple electrokinetic injections is demonstrated as a general method for inducing on capillary reactions between liposomes and other reagents. Furthermore, some preliminary evidence on the use of liposomes as a hydrophobic partitioning medium for analytical separations is presented. PMID- 21619278 TI - An analytical method for trifluoroacetic Acid in water and air samples using headspace gas chromatographic determination of the methyl ester. AB - An analytical method has been developed for the determination of trace levels of trifluoroacetic acid (TFA), an atmospheric breakdown product of several of the hydrofluorocarbon (HFC) and hydrochlorofluorocarbon (HCFC) replacements for the chlorofluorocarbon (CFC) refrigerants, in water and air. TFA is derivatized to the volatile methyl trifluoroacetate (MTFA) and determined by automated headspace gas chromatography (HSGC) with electron-capture detection or manual HSGC using GC/MS in the selected ion monitoring (SIM) mode. The method is based on the reaction of an aqueous sample containing TFA with dimethyl sulfate (DMS) in concentrated sulfuric acid in a sealed headspace vial under conditions favoring distribution of MTFA to the vapor phase. Water samples are prepared by evaporative concentration, during which TFA is retained as the anion, followed by extraction with diethyl ether of the acidified sample and then back-extraction of TFA (as the anion) in aqueous bicarbonate solution. The extraction step is required for samples with a relatively high background of other salts and organic materials. Air samples are collected in sodium bicarbonate-glycerin-coated glass denuder tubes and prepared by rinsing the denuder contents with water to form an aqueous sample for derivatization and analysis. Recoveries of TFA from spiked water, with and without evaporative concentration, and from spiked air were quantitative, with estimated detection limits of 10 ng/mL (unconcentrated) and 25 pg/mL (concentrated 250 mL:1 mL) for water and 1 ng/m(3) (72 h at 5 L/min) for air. Several environmental air, fogwater, rainwater, and surface water samples were successfully analyzed; many showed the presence of TFA. PMID- 21619279 TI - Preparation of continuous beds derivatized with one-step alkyl and sulfonate groups for capillary electrochromatography. AB - A simple one-step procedure for the preparation of columns for capillary electrochromatography is described. The nonpolar compounds (stearyl or butyl methacrylate) used for the introduction of hydrophobic ligands (C18, C4) are rendered water-soluble in the aqueous monomer mixture used for the synthesis of the matrix by the addition of a surfactant. This solution is sucked into a piece of fused silica tubing, the inner walls of which are activated by methacryl groups. Following polymerization, the continuous bed column is ready for use. Since the bed is attached covalently to the tubing wall, no frit to support the bed is required, which simplifies the preparation of the column. In addition, a frit can easily become clogged and is often a site for the generation of air bubbles. Some new approaches to increase the resolution and shorten the analysis times in electrochromatography are suggested and demonstrated experimentally, such as sharpening the starting zone, employing gradient elution, or adding SDS (below cmc) to the mobile phase. Separation of five polycyclic aromatic hydrocarbons can be achieved in 5 min in a 25-MUm-i.d. column. PMID- 21619280 TI - Application of compact porous disks for fast separations of biopolymers and in process control in biotechnology. AB - Production and downstream processing in biotechnology requires fast and accurate control of each step in the process. Improved techniques which can be validated are required in order to meet these demands. For these purposes, chromatographic units containing compact porous disks for fast separation of biopolymers were developed and investigated with regard to their performance and speed. The problems that have, in the past, arisen from the use of wide and flat separation units, such as membranes and disks, have chiefly been those of sample distribution and large void volumes before and behind the unit. Improvements in the construction of the cartridge have led to better performance of the compact porous disks and faster separation. Using these disks, three calibration standard proteins could be separated within less than 1 min by an anion-exchange, cation exchange, and hydrophobic interaction mode. Such units can be used for in-process control in production and downstream processing of biopolymers, as was shown in experiments involving the purification of alpha(1)-antitrypsin and clotting factor IX and the immobilization of enzyme glucose oxidase on an epoxy-activated compact porous disk. PMID- 21619281 TI - Oxidative stability and degradation mechanism of a cyclotriphosphazene lubricant. AB - Chromatographic and spectroscopic techniques were used to investigate the degradation products from bulk oxidative and thermal stability testing of a substituted tricyclophosphazene high-temperature lubricant. Analytical investigation into the degraded lubricant revealed oligomerization to be the dominant mode of degradation, resulting in most of the observed viscosity increases. GC/MS, FT-IR, and (31)P NMR data confirmed the existence of a cyclotetramer structure in the degraded fluid, while GPC analysis indicated the possibility of a much smaller amount of higher oligomers. The poor reproducibility observed for the oxidation results is likely due to the influence of trace contaminants or the relative degree of retention of ionizable volatile products that could act as oligomerization catalysts. PMID- 21619282 TI - Mechanism of signal suppression by anionic surfactants in capillary electrophoresis-electrospray ionization mass spectrometry. AB - Micellar-mediated capillary electrophoresis (CE) is used for a wide variety of applications, including the separation of pharmaceuticals, environmental contaminants, illicit drugs, DNA fragments, and many other biological samples. The electrospray ionization interface is one of the most common CE-MS interfaces. Coupling micellar-mediated CE separations with MS detection combines two very powerful, widely applicable analytical techniques. Some types of surfactants strongly interfere with electrospray ionization mass spectrometric (ESI-MS) detection of analytes, and in many cases the ESI-MS analyte signals are completely quenched. Only a few reports have appeared that describe the ESI-MS detection of analytes in the presence of surfactants; however, the exact mechanism of ionization suppression has not yet been addressed. In this work, a modified aerosol ionic redistribution (AIR) model is presented that qualitatively explains the results of previous studies, including those using "polymeric surfactants". Analyte ionization suppression by surfactants appears to be caused by Coulombic interaction between oppositely charged solute and surfactant ions in the ESI-produced offspring droplets. It appears that the ability of surfactants to quench electrospray ionization is directly related to the surface activity and the charge of the surfactant. Also, highly surface active components tend to be enriched in ESI-produced offspring droplets. Analyte ion signals can be detected under conditions that lower the surface concentration of oppositely charged surfactant ions in aerosol droplets. The mechanistic information outlined here may be used to design micellar-mediated CE separations that allow detection of analyte ions by ESI-MS. PMID- 21619283 TI - Octadecylsilylated porous silica rods as separation media for reversed-phase liquid chromatography. AB - Continuous porous silica rods consisting of a mesoporous (pore size, 14 or 25 nm) silica skeleton of ~1 MUm size and through-pores of ~1.7 MUm were prepared and derivatized to C(18) phase by on-column reaction with octadecyldimethyl-(N,N diethylamino)silane. The C(18) silica rods gave plate heights of 10-20 MUm for aromatic hydrocarbons in 80% methanol and 20-40 MUm for insulin in acetonitrile water mixtures in the presence of trifluoroacetic acid. The performance of the silica rods was much better at a high flow rate than that of conventional columns packed with 5 MUm C(18) silica particles having 12 and 30 nm pores, especially for high molecular weight species. PMID- 21619284 TI - A Cryotrap Membrane Introduction Mass Spectrometry System for Analysis of Volatile Organic Compounds in Water at the Low Parts-per-Trillion Level. AB - Detection of volatile organic compounds (VOCs) in aqueous solution at low parts per-trillion (ppt) levels is accomplished using a very simple and efficient on line preconcentration cryotrap membrane introduction mass spectrometry (CT-MIMS) system. The conventional MIMS probe is modified so that the membrane interface is placed about 15 cm away from the ion source. A U-shaped trap tube is then inserted between the membrane interface and the ion source. Cryotrapping is performed with liquid nitrogen for 15 min, followed by fast heating at ~15 degrees C s(-)(1), which thermally releases the condensed VOCs almost at once into the ion source region of a quadrupole mass spectrometer. By applying electron ionization and a selective ion monitoring scan mode, a very sharp and intense peak is obtained. The performance of the CT-MIMS system was compared with that of conventional MIMS, and after reaching the best conditions for the trapping and heating cycles, an improvement factor in signal intensity of about 100 was observed for a series of VOCs. The extraordinary sensitivity of CT-MIMS system allows VOCs to be detected at very low concentrations, detection limits being typically on the order of 10-20 ppt. The results also show excellent linearity and reproducibility for the system. PMID- 21619285 TI - Conversion of O(2) into CO(2) for High-Precision Oxygen Isotope Measurements. AB - An improved procedure of (18)O/(16)O ratio measurements by means of oxygen conversion to CO(2) is developed, which allows one to obtain the true delta(18)O values with a precision of +/-0.050/00 in oxygen samples down to 7 MUmol. The isotopic exchange between quartz glass and oxygen gas was measured in the temperature range of 600-900 degrees C, and it was found to be less than 0.2%. PMID- 21619286 TI - Fluorescence detection of flavonols in HPLC by postcolumn chelation with aluminum. AB - Flavonols are dietary antioxidants which may prevent coronary heart disease. To be able to study absorption of flavonols in humans, we developed a postcolumn derivatization with aluminum for HPLC with fluorescence detection. Variables governing postcolumn chelation, such as water content, buffer, organic modifier of the eluent, concentration of Al(3+), presence of acetic acid in the postcolumn reagent, and temperature, were studied and optimized. Of the flavonoids, only flavonols that contain a free 3-hydroxyl and 4-keto oxygen binding site form fluorescent complexes with Al(3+). The method has a detection limit of 0.15 ng/mL for quercetin, 0.05 ng/mL for kaempferol, 0.45 ng/mL for myricetin, and 0.05 mg/mL for isorhamnetin, thus improving detectability of quercetin 300-fold as compared to that possible with UV detection. The reproducibility relative standard deviation of the method is 1.4%. This extremely sensitive method enables, for the first time, determination of flavonols in body fluids after consumption of a normal diet. PMID- 21619287 TI - Solubility parameters and solubilities of metal dithiocarbamates in supercritical carbon dioxide. AB - The solubilities of Cu, Hg, and Zn complexes with seven different dithiocarbamate ligands in supercritical fluid CO(2) at 60 degrees C and two pressures (100 and 230 atm) are reported. In each metal chelate system, the solubility of the metal dithiocarbamate complex shows a strong correlation with the solubility parameters of the ligands, calculated using a group contribution method. Dithiocarbamate ligands with smaller solubility parameter values form metal complexes with higher solubilities in supercritical CO(2). The solubility parameter value may provide a general guideline for selecting effective ligands for metal extraction in supercritical CO(2). PMID- 21619288 TI - Fluorescence imaging of the heterogeneous reduction of oxygen. AB - The reduction of oxygen at a variety of solid electrodes was spatially imaged using fluorescence microscopy. Hydroxide produced during electrolysis of oxygen converted the acid-base indicator, fluorescein, into its fluorescent form. Fluorescence intensity was collected as a function of potential at platinum, silver, and glassy carbon disk electrodes and tracked the faradaic current due to oxygen reduction at platinum electrodes. The ability to observe spatial variations in electron-transfer kinetics was demonstrated at a bimetallic electrode prepared from silver and platinum. Fluorescence imaging of oxygen reduction on silver electrodeposited on glassy carbon revealed the location and size of the silver deposits. Imaging of oxygen reduction at a ruthenium-graphite composite electrode demonstrated the ability to identify electrochemically active sites on a spatially complex surface. PMID- 21619289 TI - Microderivatization of anodized glassy carbon. AB - Microelectrodes have been used to modify locally the electrochemical activity on glassy carbon electrodes. Glassy carbon was electrochemically oxidized to form an oxide layer which is inhibitory toward certain electron-transfer reactions. Activity was restored through the application of hydroxide, which was generated electrochemically at the tip of a microelectrode. With the tip positioned in close proximity to the anodized glassy carbon surface, microdomains of electrochemical activity were created in an otherwise inactive matrix. The distribution of electrochemical activity was characterized using electrochemical feedback at the microelectrode, electrogenerated chemiluminescence imaging, and electrodeposition of silver. Spatially directed activation of the glassy carbon surface was accomplished in the micrometer domain. PMID- 21619290 TI - Inert metal-modified, composite ceramic-carbon, amperometric biosensors: renewable, controlled reactive layer. AB - A new type of sol-gel-derived, inert metal-modified, composite, amperometric biosensor is developed. The electrodes are comprised of a dispersion of biochemically and chemically modified graphite powder in a porous, organically modified silicate (Ormosil) network. The percolating carbon dispersion provides electrical conductivity, oxidoreductase enzymes (e.g., glucose oxidase, lactate oxidase, or l-amino acid oxidase) are used for biocatalysis, metallic palladium is used for electrocatalysis of the biochemical reaction product, and the porous organically modified silica provides a rigid skeleton. The hydrophobicity of this composite material guarantees that only a limited section of the electrode is wetted by the aqueous analyte, thus providing a controlled-thickness reactive layer. The thickness of the reaction layer can be tuned by the addition of hydrophilic components. The electrode can be reproducibly renewed by removing its upper layer and exposing a new, thin, porous bioreactive section. The same technology is applicable for the production of thick-film, low-leaching, disposable sensors. In this configuration, the analysis is conducted using a single drop of the analyte applied on the hydrophobic film. The sensors are found to be stable over long periods. PMID- 21619291 TI - Ultrahigh-speed calculation of isotope distributions. AB - This paper introduces an ultrahigh-speed algorithm for calculating isotope distributions from molecular formulas, elemental isotopic masses, and elemental isotopic abundances. For a given set of input data (molecular formula, elemental isotopic masses, and elemental isotopic abundances), and assuming round-off error to be negilgible, the new algorithm rigorously produces isotope distributions whose mean and standard deviation are "correct" in the sense that an error-free algorithm would produce a distribution having the same mean and standard deviation. The peak heights are also "correct" in the sense that the height of each nominal isotope peak from the ultrahigh-speed calculation equals the integrated peak area of the corresponding nominal isotope peak from an exact calculation. As a consequence of these properties, the algorithm generally places isotope peaks within millidaltons of their true centroids. The method uses Fourier transform methods and relates closely to two other recently introduced algorithms. The suite of capabilities provided by these three algorithms is sufficient to solve an extremely wide range of problems requiring isotope distribution simulation. PMID- 21619292 TI - Enhanced Signal-to-Background Ratios in Voltammetric Measurements Made at Diamond Thin-Film Electrochemical Interfaces. AB - Large signal-to-background (S/B) ratios for the Fe(CN)(6)(3)(-)(/4)(-) and IrCl(6)(2)(-)(/3)(-) redox couples in KCl have been observed in cyclic voltammetric measurements made at a conductive diamond thin-film electrode without any conventional surface pretreatment. The S/B ratios were a factor of ~16 and 8 larger at diamond than at freshly polished glassy carbon (GC) for Fe(CN)(6)(3)(-)(/4)(-) and IrCl(6)(2)(-)(/3)(-), respectively. The polycrystalline diamond film, grown on a p-Si(100) substrate, possessed significant cubic {100} faceting, as evidenced by AFM images, and was of high quality, as indicated by Raman spectroscopy. The high degree of electrochemical activity without surface pretreatment, the enhanced S/B ratios, and the excellent response stability demonstrate that diamond might be an attractive new electrode material for electroanalysis. PMID- 21619293 TI - Conductivity and voltammetry in liquid and supercritical halogenated solvents. AB - A previous study of the voltammetry of ferrocene in liquid and supercritical chlorodifluoromethane revealed electrochemically reversible behavior. However, shifts in the half-wave potential as a function of fluid conditions were observed which were tentatively attributed to ohmic distortion due to changes in fluid resistance. To more completely understand the voltammetry in this fluid, conductivity measurements have been made for a range of fluid conditions. Additionally, a second reference couple, cobaltocenium hexafluorophosphate, has been introduced, and the difference in half-wave potentials between the two redox couples has been examined as a function of fluid conditions. In the liquid, the difference in the half-wave potentials of the two couples corrected for ohmic distortion decreases as the fluid temperature increases (25-85 degrees C) at constant pressure (5.2 MPa). In the supercritical fluid at constant temperature (115 degrees C), the difference in the half-wave potentials corrected for ohmic distortion is constant at 1.276 +/- 0.005 V over a considerable range of fluid pressure (10-30 MPa). Ion aggregation in the supercritical fluid is indicated both by the conductivity measurements and by the rather large hydrodynamic radius of cobaltocenium computed from the voltammograms. Preliminary voltammetry in supercritical trifluoromethane is also presented. PMID- 21619294 TI - A pulse amperometric sensor for the measurement of atmospheric hydrogen peroxide. AB - Gaseous H(2)O(2) is sampled through a Nafion membrane diffusion scrubber while 1 mM HCl is maintained stationary in the scrubber. After a preselected preconcentration time (typically, 5-10 min), a valve is opened to allow the scrubber liquid to flow by gravity over an electrochemical H(2)O(2) sensor for a brief period. The miniature flow-over sensor consists of a Pt/Rh wire working electrode and a Pt wire counter electrode wound respectively on separate segments of a Nafion solid polymer electrolyte tubing supported on a Ag/AgCl wire reference electrode. A simple electronic interface and a personal computer are used to control and record the electrochemical measurement. The liquid phase detection limit for this sensor is ~30 nM H(2)O(2) in the anodic oxidation mode. For a 9 min gas sample preconcentration period, the LOD (S/N = 3 criterion) is 0.11 ppbv H(2)O(2)(g). Ambient H(2)O(2) data obtained with this instrument were in excellent agreement with those obtained by an established fluorometric technique in a blind intercomparison. PMID- 21619295 TI - Gas Sensing Based on a Nonlinear Response: Discrimination between Hydrocarbons and Quantification of Individual Components in a Gas Mixture. AB - A novel sensing system is proposed based on the multidimensional information contained in a dynamic nonlinear response. A sinusoidal temperature change was applied to a SnO(2) semiconductor gas sensor, and the resulting output conductance of the sensor was analyzed by fast Fourier transformation (FFT). The higher harmonics of the FFT characterized the nonlinear properties of the response. The amplitudes of the higher harmonics of the FFT exhibit characteristic changes which depend on the chemical structure, concentration, and the kinetics of adsorption and the reaction of hydrocarbon gases and aromatic vapors on the sensor surface. In addition, it is possible to distinguish between gases in a gaseous mixture with a single detector using this dynamic nonlinear response. Nonlinear responses are discussed in relation to the kinetics of the reaction at the sensor surface and the temperature-dependent barrier potential of the semiconductor. PMID- 21619296 TI - Analysis of polar organic compounds using charge exchange ionization and membrane introduction mass spectrometry. AB - Charge exchange ionization in conjunction with membrane introduction mass spectrometry provides a sensitive method for the detection of polar volatile organic compounds and semivolatile compounds in air. Sample introduction into an ion trap mass spectrometer was accomplished with a hollow fiber silicone membrane assembly. Atmospheric oxygen, which diffuses through the membrane, was used as the charge exchange reagent. Chemical ionization parameters were optimized using methyl ethyl ketone (2-butanone) standards in air. Several other oxygen containing compounds, including acetone (2-propanone), methyl isobutyl ketone (4 methyl-2-pentanone), propanal, isopropyl alcohol (2-propanol), cyclohexanol, dimethyl sulfoxide (sulfinylbismethane), 2-(diethylamino)ethanol, and dimethyl methylphosphonate were analyzed with this technique. This method was used to obtain mass spectra for a variety of classes of compounds and produced a 4-20 fold improvement in response for all of the polar compounds we examined when compared to signal obtained from electron ionization. PMID- 21619297 TI - Radio frequency glow discharge mass spectrometry for the characterization of bulk polymers. AB - A radio frequency (rf)-powered glow discharge (GD) atomization/ionization source is utilized to determine the applicability of the technique for direct polymer analysis. A series of PTFE-based polymers are studied to assess their fingerprint mass spectra and to distinguish each sample by its differing base peaks and relative peak intensities. A parametric study with respect to discharge gas pressure and rf power is conducted to evaluate their respective roles in the sputtering process as well as possible ionization mechanisms. The results of the GD sputtering processes are examined by scanning electron micrographs of a sputtered PTFE surface. Excellent discharge stabilization characteristics (<3 min) were observed in temporal response curves. Internal stability with respect to signal intensity is observed to be <5% RSD for samples of different thicknesses. Finally, the ability to obtain depth profiles of layered samples was demonstrated for the case of a Cu layer deposited on a PTFE substrate. PMID- 21619298 TI - Rare Earth oxide equilibria in pulsed direct current glow discharge mass spectrometry. AB - Glow discharge mass spectrometry is used to examine the equilibria existing between La(+) and LaO(+). A pulsed discharge permitted temporal comparison of spectra taken at varying intervals after discharge initiation. Postdischarge peaks are observed for both atom and oxide ions. By varying the pulse period while sustaining a fixed "on" time, the degree of deposition of gaseous constituents on the cathode surface can be controlled. Injection of normal water and isotopically labeled water for compacted and noncompacted samples allows insight into the source of water signals. PMID- 21619299 TI - Numerical solution of the complete mass balance equation in chromatography. AB - A new approach to simulate the movement of bands through a chromatographic column is presented. Similar to the Craig distribution model, the mass balance equation is divided into two equations describing two successive processes. The first equation includes two effects: solute diffusion in the mobile phase and migration of the solute band with the mobile phase. The second equation deals with the distribution of the solute between phases, i.e., the adsorption isotherm. The partial differential equations are integrated numerically over time and space using two methods. The first approach is a finite difference method. In the second approach, the propagation operator is expanded in a Chebyshev series, where large time steps can be used. The rate of adsorption and desorption is determined by the size of the time increment. By varying the size of the time step, it is possible to study kinetic effects. The influences of sample size, injection width, rate of mass transfer, and mobile phase velocity on the elution profile were studied. Simulations using the modified Craig approach with either of the two numerical procedures showed that the solutes behaved in the chromatographically expected manner. Moreover, with linear adsorption isotherms, direct relationships between HETP, as well as retention times, and experimental parameters could be established. PMID- 21619300 TI - Analysis of inorganic species by capillary electrophoresis-mass spectrometry and ion exchange chromatography-mass spectrometry using an ion spray source. AB - Mixtures of inorganic ions separated by capillary electrophoresis (CE) and ion exchange chromatography (IC) are detected by mass spectrometry (MS) using an ion spray atmospheric pressure ionization source. The selectable degree of ion-adduct declustering and molecular fragmentation in the MS interface region allows the system to be operated as an elemental analyzer or as a molecular detector suitable for oxidation state determinations. Both inorganic anions and cations (including alkalis, alkaline earths, transition metals, and lanthanides) are analyzed by CE-MS. A variety of CE separation buffers are evaluated for the cation analyses (e.g., creatinine, ammonium acetate, and tris[hydroxymethyl]aminomethane). Only one of the buffers (i.e., creatinine) can be used for CE-indirect UV detection. A CE capillary permanently coated with strong anion exchange sites and a pyromellitic acid buffer (suitable for indirect UV detection) is used for the inorganic anion separations. The coated column eliminates the need for buffer modifiers to reverse the flow in the capillary, which then reduces background noise and mass spectral complexity. The separation and detection of 13 inorganic anions are also accomplished by IC using an anion exchange column with a carbonate-bicarbonate mobile phase, on-line suppressed conductivity detection, and mass spectrometric detection. PMID- 21619301 TI - Capillary biosensor for glutamate. AB - We have developed and characterized a novel glutamate biosensor which allows biological transduction of glutamate signal during transport of analyte from the sampling site to the detector. This biosensor exploits the high surface-to-volume ratio found in small-diameter fused silica capillaries. Glutamate dehydrogenase (GDH) was attached to the inner surface of a 75 MUm i.d. capillary using biotin avidin chemistry. In the presence of excess nicotinamide adenine dinucleotide (NAD(+)), GDH converts glutamate to alpha-ketoglutarate while simultaneously reducing NAD(+) to NADH. Detection of NADH was accomplished using laser-induced fluorescence. Perfusion with 30 MUM glutamate in the presence of 3 mM NAD(+) resulted in a strong increase in fluorescence, with a response time of 450 ms. This effect was abolished upon exclusion of NAD(+) from the buffer. The limit of detection is 3 MUM (S/N = 3), with a linear working range from 3 to 300 MUM. Efficiency of the GDH-modified capillary ranged between 20% and 92% and was positively correlated with concentration of glutamate. The effect of linear velocity was also examined and was shown to be indirectly related to efficiency, with maximum response observed at 4.5 cm/min. In summary, we have demonstrated the successful attachment of glutamate dehydrogenase to the inner wall of a small diameter fused silica capillary while retaining enzymatic activity. The resulting biosensor exhibits characteristics amenable for in vivo applications. Future efforts will be directed toward the incorporation of this biosensor into current technologies, such as capillary electrophoresis and microdialysis. PMID- 21619302 TI - Characterization of a range of alkyl-bonded silica HPLC stationary phases: correlation of quantitative surface analysis data with the retention behavior of neutral, acidic, and basic solutes. AB - An investigation was made of the correlation between quantitative surface analytical data obtained by XPS and static SIMS and the chromatographic performance of a range of n-alkyl-bonded silica (C1-C18) packing materials. A series of acidic, basic, and neutral solutes was used to study the retention behavior. For comparison, analysis of bulk total percentage carbon (%C) and alkyl surface density of the bonded silica particulates were also included. Significant correlations were observed, in the majority of cases, between the retention factor (k) and the XPS C:Si atomic ratio, which was similar to that obtained between k and the bulk %C or k and the bonded alkyl chain length. Similar significant correlations were also obtained between k and the static SIMS alkyl:Si ion peak area ratios. XPS alkyl:Si atomic ratios were calculated as an estimate of alkyl surface coverage of the silica support, and these correlated well with the surface density calculated from the bulk %C and the surface area of the packing material. The XPS alkyl:Si ratio also demonstrated a significant correlation with the peak asymmetry factor derived for basic solutes. These studies confirm that both XPS and static SIMS can generate surface chemical data from chromatography particulates, which has direct relevance to the prediction of chromatographic behavior. We believe that these techniques will prove to be effective tools to assist in the characterization of chromatographic supports and stationary phases. PMID- 21619303 TI - "Entropically driven" chiral separations in supercritical fluid chromatography. Confirmation of isoelution temperature and reversal of elution order. AB - Chiral separations depend upon column efficiency and chiral selectivity. Supercritical fluid chromatography (SFC) has been shown to have pronounced advantages in chiral separations due to its enhanced column efficiency at normal flow rates. Examination of factors affecting selectivity in SFC is crucial to systematic chiral method development. Selectivity is a compromise between differences in enantiomeric binding enthalpy and disruptive entropic effects. Increased temperature will decrease the effect of differences in enantiomer binding enthalpy, eventually decreasing selectivity to a point where the enantiomers coelute. Extension of this thermodynamic theory predicts that further increases in temperature lead to selectivity values of less than 1.0, and elution order of the enantiomers reverses. In this region separations are said to be "entropically driven", and selectivity increases (decreases from 1.0) with temperature. Performing separations in this region is attractive because column efficiency is also expected to increase with temperature. Such entropically driven separations have been observed only in gas chromatography. Most data used to support the application of this theory in SFC have been generated at subcritical temperatures, while theory purports to predict behavior above the critical temperature (T(c)). This approach ignores the effects of traversing the critical temperature in SFC, which is known to have a variety of unpredictable consequences. Work presented here shows the effect of temperatures above T(c) on chromatographic behavior. Contrary to theory, capacity factors increase near T(c) and column efficiency declines. Use of pressures well above the critical pressure lessens these effects. In accordance with theory, selectivity does decrease with temperature through T(c) and isoelution temperatures and two instances of elution order reversal are observed here for the first time in SFC. PMID- 21619304 TI - Determination of total organic carbon in water by thermal combustion-ion chromatography. AB - A sensitive method for determining total organic carbon at microgram per liter levels in industrial, environmental, and drinking waters by thermal combustion ion chromatography was developed using tube furnace and readily accessable HPLC equipment. To achieve complete oxidation, persulfate (0.25%) was added to oxidize nonvolatile organic compounds in solution and cupric oxide heated at 900 degrees C to convert volatile organic compounds to CO(2), which was scrubbed in a 20 mL solution of 50 mM KOH with 10 drops of butanol added. The carbonate anion obtained was determined by nonsuppressed ion chromatography using 0.6 mM potassium hydrogen phthalate (KHP) as the eluent. Both surfactants and volatile and nonvolatile organic compounds commonly found in environmental waters give highly repeatable recoveries close to 100%. The detection limit (S/N = 2) and linear range for a 1 L water sample are 2 MUg of C L(-)(1) and 10-2500 MUg of C L(-)(1), respectively, and they can be adjusted using samples ranging from 100 mL to 2 L. Good repeatablity (RSD less than 10%) and close to 100% recoveries were obtained for KHP added to real samples such as deionized, mineral, tap, and river water and seawater. Compared with the ASTM D2579 method, the method developed is 3 orders of magnitude more sensitive, more accurate, and reliable in determining samples with low total organic carbon values and more flexible in adjusting the linear range and sensitivity using variable sample sizes. PMID- 21619305 TI - Rapid analyte recognition in a device based on optical sensors and the olfactory system. AB - We report here the development of a new vapor sensing device that is designed as an array of optically based chemosensors providing input to a pattern recognition system incorporating artificial neural networks. Distributed sensors providing inputs to an integrative circuit is a principle derived from studies of the vertebrate olfactory system. In the present device, primary chemosensing input is provided by an array of fiber-optic sensors. The individual fiber sensors, which are broadly yet differentially responsive, were constructed by immobilizing molecules of the fluorescent indicator dye Nile Red in polymer matrices of varying polarity, hydrophobicity, pore size, elasticity, and swelling tendency, creating unique sensing regions that interact differently with vapor molecules. The fluorescent signals obtained from each fiber sensor in response to 2-s applications of different analyte vapors have unique temporal characteristics. Using signals from the fiber array as inputs, artificial neural networks were trained to identify both single analytes and binary mixtures, as well as relative concentrations. Networks trained with integrated response data from the array or with temporal data from a single fiber made numerous errors in analyte identification across concentrations. However, when trained with temporal information from the fiber array, networks using "name" or "characteristic" output codes performed well in identifying test analytes. PMID- 21619306 TI - Nondestructive Determination of Major Elements in a Large Sample by Prompt gamma Ray Neutron Activation Analysis. AB - An internal monostandard method is proposed for the nondestructive determination of major elements in large samples by prompt gamma ray neutron activation analysis. It successfully overcomes the problems characteristic of large samples, such as the absorption and scattering of incident neutrons and the absorption of emitted gamma rays. In order to make this proposed method understood theoretically, an equation is presented and its validity for the analysis of large samples discussed. In principle, the proposed method gives relative contents in large solid samples, whereas it allows absolute determination for samples in solution form. As a demonstration for the analysis of a large solid sample, we analyzed an earthen vessel, and the major elements in the sample (Na, Mg, Al, Si, K, Ca, Ti, Mn, and Fe) were determined within the uncertainty of 10%, except for Mn. This method was also tested for samples in solution form, and it was found that the absolute content of a target element could be obtained by constructing a calibration curve using several known standard solutions of different concentrations. Residual radioactivity after irradiation was also examined and found to be so little that the sample could be taken outside the radiation-controlled area within a few days after the irradiation. PMID- 21619307 TI - Performances of mass-sensitive devices for gas sensing: thickness shear mode and surface acoustic wave transducers. AB - In this work we investigated different thickness shear mode resonators (TSMRs) with fundamental frequencies of 10 and 30 MHz and surface acoustic wave devices with fundamental frequencies of 80 and 433 MHz. Four aspects were of primary interest in this comparison: noise levels and signal-to-noise ratios (S/N), influence of the polymer film thickness, influence of temperature on the transducer signal before and after coating, and minimum threshold values for monitoring different volatile organic compounds in the environment. We limited our investigations to a temperature range between 298 and 308 K, with 303 K the routine measuring temperature. Analyte concentrations (n-octane, tetrachloroethene) were chosen from the minimum detection limit up to 5000 MUg/L. The temperature was found to strongly affect the performance of all the devices. The sorption of the analyte vapors into the polymeric films was demonstrated to be transducer-independent (identical partition coefficients for all the devices). The 30 MHz TSMRs showed very satisfying results in terms of S/N and limits of detection. PMID- 21619308 TI - QCM Operation in Liquids: An Explanation of Measured Variations in Frequency and Q Factor with Liquid Conductivity. AB - Recently, several reports have shown that when one side of a quartz crystal microbalance (QCM) is exposed to a liquid, the parallel (but not the series) resonant frequency is influenced by the conductivity and dielectric constant of the liquid. The effect is still controversial and constitutes a serious complication in many applications of the QCM in liquid environments. One suggestion has been that acoustically induced surface charges couple to charged species in the conducting liquid. To explore this effect, we have measured the parallel and the series mode resonance frequencies, and the corresponding Q factors, for a QCM with one side facing a liquid. These four quantities have all been measured versus liquid conductivity, using a recently developed experimental setup. It allows the simultaneous measurement of the resonant frequency and the Q factor of an oscillating quartz crystal, intermittently disconnected from the driving circuit. Based on these results, a simple model together with an equivalent circuit for a quartz crystal exposed to a liquid is presented. The analysis shows that it is not necessary to infer the existence of surface charges (or other microscopic phenomena such as electrical double layers) to account for the influence of the liquid's electrical properties on the resonant frequency. Our results show that the contacting conductive liquid, in effect, enlarges the electrode area on the liquid side and thereby changes the parallel resonant frequency. By proper design of the QCM measurement, perturbing effects due to the liquid's electrical properties can be circumvented. PMID- 21619309 TI - Electrical Double-Layer Effects and the Origin of Indirect UV Detection in the Liquid Chromatography of a Cationic Sample Using a Cationic Probe. AB - Two cations, tetra-n-butylammonium (TBA(+)) and (4-nitrobenzyl)trimethylammonium (NBTA(+)), are simultaneously sorbed on the bonded phase sorbent Partisil-10 ODS 3 from water at various ionic strengths. The non-UV-absorber TBA(+) is thought of as the sample ion and the UV-absorber NBTA(+) as the probe ion in an indirect detection system. In the sorbed state, the two ions reside at different charge planes: NBTA(+) is very close to the ODS-water interface, and TBA(+) is about 11 A deeper into the ODS phase. Both ions contribute to the electrical potential psi(I) at the NBTA(+) charge plane, which can be calculated from an electrical triple-layer model. The sorption of TBA(+) influences the amount of NBTA(+) sorbed both by raising psi(I) and by altering the amount of space available for NBTA(+). The former effect is greater than the latter at ionic strengths up to at least 0.5 mol/L. For the latter effect, the direction and magnitude of the influence of TBA(+) on available space both change with ionic strength (c). At low c, sorbed TBA(+) increases the amount of space available for NBTA(+) by disrupting the conformation of the alkyl chains of the ODS phase. At high c, sorbed TBA(+) decreases the amount of space available by competing with NBTA(+) for space. This dramatically different effect of TBA(+) on available space at low and high c occurs because higher c produces both a weaker interaction of the ODS alkyl chains with water and a reorientation of NBTA(+) from flat to perpendicular in the ODS phase. PMID- 21619310 TI - Solvent microextraction into a single drop. AB - An analytical technique is described which combines solvent extraction with gas chromatographic (GC) analysis in a simple and inexpensive apparatus involving very little solvent consumption. A small drop (8 MUL) of a water-immiscible organic solvent, containing an internal standard, is located at the end of a Teflon rod which is immersed in a stirred aqueous sample solution. After the solution has been stirred for a prescribed period of time, the probe is withdrawn from the aqueous solution, and the organic phase is sampled with a microsyringe and injected into the GC for quantification. The observed rate of solvent extraction is in good agreement with a convective-diffusive kinetic model. Analytically, the relative standard deviation of the method is 1.7% for a 5.00 min extraction of the analyte 4-methylacetophenone into n-octane. PMID- 21619311 TI - Application of Eigenstructure Tracking Analysis and SIMPLISMA to the Study of the Protonation Equilibria of cCMP and Several Polynucleotides. AB - The application of eigenstructure tracking analysis (ETA) and SIMPLISMA for the investigation of the protonation equilibria of a monomer and several polynucleotides is proposed. Both approaches have been applied in the pH and in the wavelength direction to the spectroscopic data matrices obtained in the study of each equilibrum. ETA provides information about the number of components in the system, their evolution along the titration, and the local rank. SIMPLISMA is also used to obtain the number of compounds in the system, the concentration profiles, and the unit spectrum of each compound. The results obtained with SIMPLISMA and those obtained previously with the alternating least-squares approach are compared. PMID- 21619312 TI - DNA biosensor for the detection of hydrazines. AB - A double-stranded (ds) DNA-coated carbon paste electrode is employed as a remarkably sensitive biosensor for the detection of hydrazine compounds. The sensor relies on monitoring changes in the intrinsic anodic response of the surface-confined DNA resulting from its interaction with hydrazine compounds and requires no label or indicator. Short reaction times (1-10 min) are sufficient for monitoring part-per-billion levels of different hydrazines. Applicability to untreated natural water samples is illustrated. The response mechanism is discussed, along with prospects of using DNA biosensors for quantitaing other important molecules and elucidating DNA interactions and damage. PMID- 21619313 TI - Characterization of a simple Raman capillary/fiber optical sensor. AB - The characterization of a simple, dual-fiber quartz capillary/fiber optical sensor (C/FOS) for remote excitation and collection of Raman signals is presented. The Raman signals acquired with the C/FOS exhibit a 70-fold sensitivity enhancement and a 50-fold improvement in detectability relative to those obtained with the corresponding conventional dual-fiber sensor without the capillary. A background spectral feature at 790 cm(-)(1) is related to the optical fiber background and is not due to the capillary tube. With no focusing lenses or filters needed at the sample site, the remote Raman C/FOS is easy to assemble and use, and it is relatively inexpensive compared to other designs. PMID- 21619314 TI - Reduction of luminescent background in ultrasensitive fluorescence detection by photobleaching. AB - In luminescence-based ultrasensitive analysis, such as single-molecule detection by flow cytometry, the luminescence background from impurities present in the solvent or reagents can ultimately determine the detection limits. A simple, versatile method for reducing luminescence background is described. The method is based on photobleaching the reagent stream immediately before it enters the detection flow cell. Dramatic reduction (an order of magnitude or more) of both low-level continuous background and single-molecule fluorescence bursts is demonstrated. Application and enhancements of the technique are discussed. PMID- 21619315 TI - Tributyl phosphate as a sensitivity-enhancing solvent for organotin in carbon furnace atomic absorption spectrometry. AB - Tributyl phosphate (TBP) has been found to be a sensitivity-enhancing solvent for organotin compounds in graphite furnace atomic absorption spectrometry; (C(4)H(9))(2)Sn(O(2)CCH(3))(2), (C(4)H(9))(2)Sn(O(2)CC(11)H(23))(2), (C(4)H(9))(3)SnCl, and (C(4)H(9))(4)Sn all give 1 order of magnitude higher sensitivities in TBP than in toluene or ethyl acetate. The sensitivities are enhanced further 1-2 orders of magnitude in TBP, when PdCl(2)(CH(3)CN)(2) is added as a matrix modifier in the organic solvent. Among the four organotin compounds, (C(4)H(9))(2)Sn(O(2)CCH(3))(2) and (C(4)H(9))(2)Sn(O(2)CC(11)H(23))(2) give better sensitivities than (C(4)H(9))(3)SnCl and (C(4)H(9))(4)Sn in the absence of palladium in any organic solvent, which suggests that the oxygen atom in the tin compound might form tin oxides that are resistant to volatilization loss during ashing. Scanning electron microscopic, electrothermal vaporization ICPMS, and powder X-ray diffraction studies show that the final products before atomization include phosphorus-containing compounds Sn(2)P(2)O(7), SnP(2)O(7), and Pd(9)P(2), besides tin-palladium alloys, PdSn, Pd(3)Sn, Pd(2)Sn, Pd(3)Sn(2), and PdSn(3). These phosphorus-containing compounds would more efficiently stabilize tin and suppress tin vaporization loss during ashing, to give higher sensitivity. PMID- 21619316 TI - Using capillary electrophoresis to follow the acetylation of the amino groups of insulin and to estimate their basicities. PMID- 21619317 TI - Applications of Frontal Affinity Chromatography to the Study of Interactions between Metal Ions and a Complex Biomaterial. AB - The use of frontal affinity chromatography for the study of metal-biomaterial interactions is described. Both normal frontal affinity chromatography and a modification of this methodology were used to generate metal binding isotherms to a biomaterial. This modification enabled the acquisition of binding isotherms with extended concentration ranges at the expense of time-dependent binding information. Comparison between normal and modified mode was made by using a well defined commercial resin. Similar performance of these two modes was obtained. The biomaterial investigated was composed of cell fragments from the plant Datura innoxia which were immobilized within a polysilicate matrix. The application of a regularized least-squares method indicated the existence of two classes of sites on this biosorbent involved in the binding of Ag(+). A total metal-ion binding affinity order at solution pH 3-5 was determined to be Cu(2+) > Cd(2+) ~ Ag(+) > Ca(2+). PMID- 21619318 TI - Retention of Ionizable Compounds on HPLC. 2. Effect of pH, Ionic Strength, and Mobile Phase Composition on the Retention of Weak Acids. AB - A new model that relates the retention of a weak acid in HPLC columns with the pH and ionic strength of the mobile phase is derived and tested for different benzoic acids in methanol-water mobile phases. The proposed model uses the pH value in the mobile phase instead of the pH value in water, takes into account the effect of the activity coefficients, and considers different holdup times for the neutral and ionic species. The dependence of the holdup time of the ionic species on the mobile phase properties (pH, solvent composition, and ionic strength) is evaluated. It is demonstrated that the holdup time of the neutral species does not depend on the mobile phase properties, but the holdup time of the ionic species depends on the particular buffer used. The proposed equations can be combined with previously derived equations that relate the retention with the solvent composition of the mobile phase to establish a general model that relates the retention of the solute with the significant mobile phase properties: composition, pH, and ionic strength. PMID- 21619319 TI - Chain order and mobility of high-density c(18) phases by solid-state NMR spectroscopy and liquid chromatography. AB - C(18) phases prepared by different synthetic pathways are examined by solid-state NMR spectroscopy. Silane functionality is clearly indicated by (29)Si CP/MAS NMR spectroscopy. Order and mobility of the alkyl chains are investigated with high speed (1)H MAS and (13)C CP/MAS NMR spectroscopy. Differences in coverage are monitored by (1)H line widths,( 13)C chemical shifts, (13)C cross-polarization constants, and (1)H relaxation times in the rotating frame. It is shown that C(18) phases prepared by the surface polymerization technique exhibit a more regular surface coverage than sorbents prepared by conventional polymeric synthesis. The findings from solid-state NMR investigations are discussed in the context of liquid chromatography (LC) separations of linear and bulky polycyclic aromatic hydrocarbon (PAH) solutes. PMID- 21619320 TI - Spectroscopic Probing of Mixed-Mode Adsorption of Ru(bpy)(3)(2+) to Silica. AB - Evanescent wave excitation of fluorescence was used to study the adsorption of Ru(bpy)(3)(2+) from aqueous solution to three types of surfaces: bare silica, a dimethylethylsiloxane (C(2)) monolayer on silica, and a dimethyloctadecylsiloxane (C(18)) monolayer on silica. The solution pH was varied to investigate the nonpolar and electrostatic contributions to the free energy of adsorption for each surface. The pH dependence of the adsorption showed that the pK(a) is the same for each of the three surfaces, consistent with earlier conclusions that the acidity of derivatized silica surfaces is due to areas of exposed silica. The free energies of adsorption for the bare silica surface, -26.2(+/-0.2) kJ/mol at pH 8, was attributed to electrostatic interactions alone. The free energy of adsorption for the C(2) and C(18) surfaces, -28.5(+/-0.1) and -31.5(+/-0.1) kJ/mol, respectively, were found to have both electrostatic and nonpolar contributions, with the latter being larger by 50% for the C(2) surface and 100% for the C(18) surface. Using Gouy-Chapman theory, the surface charge densities on each of the three surfaces, calculated from the electrostatic interaction energy of Ru(bpy)(3)(2+), were found to be within the range of literature values: 8.8(+/-0.1) * 10(-)(7) mol/m(2) for bare silica and 1.7(+/-0.1) * 10(-)(7) mol/m(2) for both the C(18) and C(2) surfaces. The results demonstrate that a cationic dye can be used to probe the silanol activity of chemically modified silica surfaces. The results support the picture that these chemically modified silica surfaces are acidic due to molecular-scale areas of contact between the bare silica substrate and the aqueous phase. PMID- 21619321 TI - Steady-state limiting currents determined by coupled diffusion, migration, and chemical equilibrium. AB - A simple theoretical model is presented for the reduction of a singly charged cation under conditions where migration is important and the cation is coupled to a neutral species through a chemical equilibrium, AB = A(+) + B(-). Only the steady-state transport-limited current, I(l), is considered. Simple algebraic equations describe the ratio of I(l) to the diffusion-limited current, I(d), as it depends on the degree of dissociation, determined by the ratio of equilibrium constant to formal concentration, K(AB)/C*(AB). The ratio I(l)/I(d) is found to depend on the ratio of electrolyte to equilibrium concentration of A(+) in bulk solution just as for the well-known result for the case without the equilibrium (i.e., K(AB) -> infinity). The results are in accord with published experimental data for weak acids. Agreement and disagreement with other theoretical treatments of this problem are discussed. The main results are for 1:1 supporting electrolytes; extensions are made to 2:1, 1:2, and 2:2 supporting electrolytes. PMID- 21619322 TI - Determination of 18 siderophile elements including all platinum group elements in chondritic metals and iron meteorites by instrumental neutron activation. AB - An instrumental neutron activation method is developed to analyze chondritic metals and iron meteorites. By changing irradiation and decay times, and selecting suitable gamma-ray and X-ray photopeaks, not only all platinum group elements (Ru, Rh, Pd, Os, Ir, Pt) but also other siderophilic elements (Fe, Co, Ni, Cu, Ga, Ge, As, Mo, Sb, W, Re, Au) can be nondestructively determined in the meteoritic metal samples. To obtain analytical data as accurate as possible, interfering reactions and neutron flux gradients during irradiation are considered. Siderophile elemental abundances measured for the Odessa iron meteorite are highly consistent with the literature values. Rh abundances for bulk H, L, LL, EH, and EL chondrites, which had been scarcely reported in the literature, are derived from Rh/Ni abundance ratios in the metal separates of the corresponding chondritic groups. PMID- 21619323 TI - Speciation analysis for organotin compounds in biomaterials after integrated dissolution, extraction, and derivatization in a focused microwave field. AB - Hydrolysis with acetic acid carried out in a low-power focused microwave field in the presence of sodium tetraethylborate and nonane is shown to shorten the sample preparation time for gas chromatographic determination of organotin compounds in biological materials. After a 3-min reaction time, ethyl derivatives of mono-, di , and tributyltin, and triphenyltin are quantitatively (>95%) found in the supernatant organic phase that is injected onto a capillary GC column. Two rapid one-step analytical procedures, using flame photometric and atomic emission detection, respectively, were developed on this basis and validated by analyzing the NIES11 certified reference fish tissue. PMID- 21619324 TI - Ferrocenylethyl phosphate: an improved substrate for the detection of alkaline phosphatase by cathodic stripping ion-exchange voltammetry. Application to the electrochemical enzyme affinity assay of avidin. AB - The ferrocenylethyl phosphate disodium salt was synthesized and used as a new substrate for alkaline phosphatase (AP). The enzyme-generated ferroceneethanol was selectively and sensitively detected at a Nafion film-coated electrode by anodic preconcentration of the ferricinium salt, followed by cyclic voltammetry. The accumulated ferricinium units could be expelled from the polymer film in their neutral form by cathodic stripping, and so the Nafion-modified electrode could be reused for more than 10 measurements with a standard deviation less than 3%. Values of 0.75 mM for the Michaelis constant and 1.42 MUmol s(-)(1) (mg of protein)(-)(1) for the maximal velocity were found. The regenerable Nafion-coated electrode was employed for the indirect detection of AP down to 2 * 10(-)(12) M and for the noncompetitive heterogeneous enzyme assay of avidin, whose detection limit was 5 * 10(-)(12) M. PMID- 21619325 TI - Anion-selective electrodes based on long-chain methyltrialkylammonium salts. AB - A new series of methyltrialkylammonium salts with an alkyl chain length (n) longer than the conventional methyltridodecylammonium (MTDDA, n = 12) has been developed, and these materials were examined for use as the ion-sensing component (ligand) in anion-selective electrodes (ISEs). Syntheses of the higher ammoniums with n = 16, 18, and 20 were carried out. In combination with an alcoholic plasticizer, the ammoniums with n = 12, 16, and 18 led to ISEs with fundamental characteristics, such as slope sensitivity, impedance, and time response, that were sufficient for practical applications. Compared with the conventional MTDDA, the ISEs based on the ligands of n = 16 and 18 showed marked improvement in chloride selectivity over both lipophilic and hydrophilic anions, deviating from the Hofmeister regime in some cases. Taking perchlorate as an example, the magnitude of the improvement was a factor of 20 for n = 16 and 15 for n = 18. When the new ISEs were applied to chloride analysis in blood serum, they improved the accuracy by a factor of 2-6. Therefore, the methyltrialkylammonium salts with alkyl chain lengths of 16 and 18 offer definite advantages over the conventional alternative and are strong candidates to become the standard compounds for use in future chloride ISEs. PMID- 21619326 TI - Ion-exchange voltammetry at polymer film-coated nanoelectrode ensembles. AB - Ensembles of nanoscopic disk-shaped electrodes have been shown to offer enhancements in electroanalytical detection limits relative to electrodes of macroscopic dimensions (e.g., disk electrodes with diameters of ~1 mm). Enhancements in electroanalytical detection limits have also been observed at macroscopic electrodes that have been coated with films of ion-exchange polymers. In this paper we combine these two concepts. We demonstrate that a nanoelectrode ensemble (NEE) that has been coated with a thin film of the Kodak ion-exchange polymer AQ 55 shows enhanced electroanalytical detection limits relative to the uncoated NEE and to the coated macroscopic electrode. To our knowledge, this is the first investigation of the electrochemistry, and the electroanalytical advantages, of polymer film-coated NEEs. PMID- 21619327 TI - Structural ion selectivity of thia crown ether compounds with a bulky block subunit and their application as an ion-sensing component for an ion-selective electrode. AB - Several 14-membered thia crown ether ionophores having a bulky block subunit were synthesized, and their chemical structures and ion selectivities were examined in detail when these compounds were used as an ion-sensing component of an ion selective electrode. The ionophores of both cyclic and noncyclic thia ethers exhibited a high selectivity for silver ion (Ag(+)), in which the sulfur atom in the ionophore molecule plays a role as the effective coordination donor site for the silver ion. The best Ag(+)-selective electrode was prepared with the 14 membered thia crown ether having one sulfur atom, three oxygen atoms, and a bulky pinan subunit. The ion selectivity of this electrode for Ag(+) was over 10(4) times that for other metal cations. In the case where the sulfide in the thia ether ionophore was changed to sulfoxide by oxidation, ion selectivity for mercury ion became higher; therefore, the sulfoxide was found to be an effective coordination site for the mercury ion. The ion selectivity features of noncyclic sulfide, sulfoxide, and sulfone were also examined and compared with the results of the cyclic and noncyclic thia ethers. PMID- 21619328 TI - Effect of diffusion coefficient diversity on steady-state voltammetry when homogeneous equilibria and migration are encountered. AB - An exact treatment is developed to predict the steady-state limiting voltammetric current, I(L), for a system in which the reaction O(+) + A(-) ? N occurs reversibly in solution, in the presence of supporting electrolyte C(+)A(-). Either or both O(+) and N undergo a one-electron reduction at the hemispherical microelectrode. The dependence of I(L) on the formation constant of the equilibration reaction, the electrolyte concentration, and the support ratio is derived for any trio of values of these parameters and for any combination of diffusivities of the O(+), N, and A(-) species. PMID- 21619329 TI - Permselectivity, Sensitivity, and Amperometric pH Sensing at Thioctic Acid Monolayer Microelectrodes. AB - Gold fiber thioctic acid (TA) monolayer (thickness of ~8 A) ultramicroelectrodes (UMEs) have been fabricated and characterized. Gold fibers were acid etched in order to produce monolayers that can act as high-quality permselective membranes. The selectivity and sensitivity of the TA monolayer UMEs compares favorably to that of TA monolayers prepared on Au vacuum deposited on large (~1 cm(2) area) single-crystal silicon wafers. The results obtained here illustrate selectivity of the monolayer electrodes to electroinactive electrolyte anions, which participate indirectly in the electrode reaction, in addition to the previously established selectivity to electroactive cations and anions. The permselectivity of the monolayer to electrolyte anions can result in lower than expected steady state currents at the monolayer UMEs, at the reaction potentials of fast-reacting analytes, and at high overpotentials for kinetically slow probes. Application of TA UMEs to amperometric pH sensing is also described in the determination of the pK(a) of TA at the monolayer surface. The method is compared with other methods of monolayer pK(a) measurements. PMID- 21619330 TI - Electrical Communication between Electrodes and Enzymes Mediated by Redox Hydrogels. AB - Different redox polymers based on poly(allylamine) with covalently attached ferrocene and pyridine groups that coordinate iron and ruthenium complexes were prepared, and hydrogels were obtained by cross-linking them with epichlorohydrin. Charge propagation from the underlying electrode, through the redox polymer and electrical communication with the enzyme FADH(2) of glucose oxidase, was studied by cyclic voltammetry and electrochemical impedance spectroscopy. The effects of electrolyte composition, concentration of enzyme and substrate, and electrode potential are reported. The role of different redox mediators covalently attached to the polymer backbone is discussed in terms of driving force and electrostatic barriers. PMID- 21619331 TI - Dual-inlet chemical amplifier for atmospheric peroxy radical measurements. AB - The state-of-the-art of peroxy radical measurements using the technique of chemical amplification has matured in recent years. The NCAR chemical amplifier has been improved over previous versions by employing a dual-inlet system that allows more rapid switching between measurement and background modes. Further improvement is realized by the use of two NO(2) detectors. The dual-inlet chemical amplifier (DICHAMP) employs two identical glass inlet reactor-NO(2) detector combinations, one of which is operated in the background, while the other operates in the radical measurement mode. This instrument is less sensitive to fluctuations in ambient ozone than the single-channel version because of the continuous monitoring of the background and background plus radical signals. The single-inlet, dual-inlet with single-detector, and DICHAMP instruments are compared through theoretical calculations of the effect of noise at a given frequency and amplitude on retrieved radical levels. Laboratory experiments were conducted to support the theoretical results. Ambient radical concentrations were determined using these configurations to evaluate the performance under actual measurement conditions. PMID- 21619332 TI - Influence of discharge parameters on the resultant sputtered crater shapes for a radio frequency glow discharge atomic emission source. AB - A parametric study has been conducted on a radio frequency glow discharge atomic emission spectrometry (rf-GD-AES) source to evaluate the sputtering characteristics and resultant crater shapes for both metallic and nonconducting samples. These studies include a determination of how the operating parameters, namely, power and pressure, influence the sputtered crater's depth, the degree of convexity and/or concavity, and the crater bottom roughness. The results imply that many similarities exist between the rf mode of powering and the well characterized dc powered devices. Some differences are noted, namely, differing crater shapes at low operating pressures and a lesser dependence of crater flatness on the applied voltage, i.e., dc-bias voltage. A new method of quantifying the degree of concavity/convexity is introduced which involves comparison of the calculated (ideal) cross-sectional area to the resultant crater area as measured by a diamond stylus profilometer. The studies indicate that the rf-GD is well suited for depth-resolved analysis under correctly chosen operating conditions, which fortuitously correspond to those used for optimum bulk elemental analyses. PMID- 21619333 TI - Environmental Analysis by ab Initio Quantum Mechanical Computation and Gas Chromatography/Fourier Transform Infrared Spectrometry. AB - Computational chemistry, in conjunction with gas chromatography/mass spectrometry/Fourier transform infrared spectrometry (GC/MS/FT-IR), was used to tentatively identify seven tetrachlorobutadiene (TCBD) isomers detected in an environmental sample. Computation of the TCBD infrared spectra was performed with the Gaussian quantum chemistry software. The Hartree-Fock/6-31G* level of theory was employed, with IR frequencies scaled by a standard factor of 0.89. This approach shows great promise as a means of characterizing or confirming environmental analyte identifications when standard spectra, or pure standards required to measure standard spectra, are unavailable. PMID- 21619334 TI - Prediction of reduced ion mobility constants of organic compounds from molecular structure. AB - Quantitative structure-property relationships (QSPRs) are used to develop mathematical models that accurately predict the reduced ion mobility constants (K(0)) for a set of 168 organic compounds directly from molecular structure. The K(0) values are taken from an unpublished database collected by G. A. Eiceman, Chemistry Department, New Mexico State University. The data were collected using a Graseby Ionics environmental vapour monitor (EVM) gas chromatography/ion mobility spectrometer. Standardized conditions with controlled temperature, pressure, and humidity were used, and 2,4-lutidine was used as an internal standard. K(0) values were measured for all monomer peaks. The best model was found with a feature selection routine which couples the genetic algorithm with multiple linear regression analysis. The set of six descriptors was also analyzed with a fully connected, feed-forward neural network. The model contains six molecular structure descriptors and has a root-mean-square error of about 0.04 K(0) unit. The descriptors in the model lend insight into some of the important molecular features that influence ion mobility. The model can be utilized for prediction of K(0) values of compounds for which there are no empirical K(0) data. PMID- 21619335 TI - On-line probe for fast electrochemistry/electrospray mass spectrometry. Investigation of polycyclic aromatic hydrocarbons. AB - A newly invented probe accessory for fast electrochemistry/electrospray mass spectrometry (EC/ESMS) is presented and evaluated. The device features a low volume, three-electrode electrochemical cell which has been designed with a minimum distance between the working electrode and the "Taylor cone" inherent to the electrospray process. This configuration limits the time between electrochemical generation of ions and mass spectrometric analysis to an absolute minimum. A fused-silica layer insulates the microcylinder working electrode from the sample solution until immediately prior to the electrospray region, postponing electrode processes until the last moment. The same fused-silica layer insulates the working electrode from the surrounding auxiliary electrode, a stainless steel capillary that also serves as the electrospray capillary. The performance and capabilities of the novel electrochemistry/electrospray mass spectrometry system have been evaluated using polycyclic aromatic hydrocarbons (PAHs) as test analytes. In the positive ion EC/ESMS mode, oxidized forms (one electron removal) of PAHs are produced in high yield. The ability to analyze reaction products appearing subsequent to the initial oxidation is also demonstrated. PMID- 21619336 TI - Ionization energy determination by the kinetic method. AB - Ionization energies of organic compounds can be determined by the kinetic method by dissociation of radical cations of van der Waals complexes. The ionized dimeric complexes of benzene and substituted benzenes, generated in the ion source of a multiquadrupole instrument by gentle charge exchange chemical ionization using carbon disulfide as reagent gas, when mass-selected and allowed to undergo collision-induced dissociation with argon, yield only the two individual radical cations as products. The logarithm of the ratio of their ion abundances correlates linearly with their ionization energies. Using this linear relationship, the ionization energy of 3-iodobenzonitrile was determined to be 9.39 +/- 0.05 eV, and this case serves to illustrate the application of the kinetic method in the determination of an unknown ionization energy. From the slope of the kinetic method plot, it is evident that the clusters are weakly bound (effective temperature, 1670 K). Strengths of this method are the simplicity of the procedure and its potential application to a wide variety of compounds. PMID- 21619337 TI - Resonance ejection from the paul trap: a theoretical treatment incorporating a weak octapole field. AB - In response to the growing experimental evidence of the importance of nonlinear phenomena in ion trap operation, a new theoretical model of ion ejection is developed. The pseudopotential well approximation for forced ion oscillations in an ion trap under the conditions of ion-molecule collisions is modified to include octapole perturbations on the quadrupole field. Ion ejection is investigated using the first-order Mitropol'skii asymptotic method for both infinitesimal and finite scan rates. It is shown that the combined action of collisional damping and nonlinearity distorts the resonance curve in such a way that "quenching" of oscillations takes place. As a result, with appropriate excitation and direction of scanning, the amplitude increases as if no damping exists! The main characteristics of the jump are derived as functions of scan rate and used for analytical estimation of mass resolution, mass peak width, and excitation voltage. Satisfactory agreement between calculated and experimental peak widths is demonstrated for the range of scanning rates in excess of 6 orders of magnitude. PMID- 21619338 TI - Reexamination of Dependence of Plate Number on SDS Concentration in Micellar Electrokinetic Chromatography. AB - Chromatograms of hydrophilic, hydrophobic, and intermediate-polarity analytes were developed in 50-MUm capillaries by micellar electrokinetic chromatography at field strengths less than 31 kV/m. The analytes were solubilized by phosphate/borate buffers containing 15, 50, and 100 mM sodium dodecyl sulfate (SDS). The plate numbers N of the analytes, as well as those of the electroosmotic flow and micellar markers, were compared to predictions of N estimated by a simple model based on longitudinal diffusion and plug size. Good to fair agreement between theory and experiment was obtained for the hydrophilic and intermediate-polarity analytes in all buffers over the entire field strength range. Good agreement between theory and experiment was obtained for the hydrophobic analyte and micellar marker in all buffers at low field strengths; however, these compounds were subject to dispersion at higher field strengths by what appears to be Joule heating. The magnitudes of other, closely related Joule heating losses are quantified here using temperature profile measurements by Morris and co-workers and Taylor dispersion calculations. In contrast to the commonly reported increase of N with media concentration, the Ns of the hydrophilic and intermediate-polarity analytes were found to be essentially independent of SDS concentration over the investigated SDS range, and the Ns of the hydrophobic species were found to be independent of SDS concentration until (what appears to be) Joule heating became significant. These results were compared to those of Sepaniak and Cole. A critique of some previous studies of N vs SDS concentration is presented, in which quantitative explanations for some dispersions are offered as alternatives to surfactant concentration effects. PMID- 21619339 TI - Hydrophobicity of cationic solutes measured by electrokinetic chromatography with cationic microemulsions. AB - The effect of the structure of employed surfactants on the selectivity of microemulsion electrokinetic chromatography (MEEKC) was investigated. Both their ionic groups and their hydrocarbon chain lengths did not affect the selectivity for neutral solutes. However, for cationic solutes, ionic interactions between catonic solutes and anionic surfactants both in the aqueous phase and in the microemulsion phase were observed. Based on these results, determination of the hydrophobicity of cationic solutes was done using MEEKC with cationic microemulsions. The obtained migration index (MI), which had been proposed as a novel hydrophobicity scale measured by MEEKC (Ishihama, Y.; et al. Anal. Chem. 1995, 67, 1588), correlated with conventional hydrophobicity scales for a water/1 octanol system, a liposome system, and an HPLC system using a phospholipid immobilized artificial membrane (IAM) column. Quantitative structure-activity relationship studies on a protein binding affinity using MI as well as the hydrophobicity scales using the liposome and the IAM column provided better results than those using the water/1-octanol system. PMID- 21619340 TI - Integrated capillary electrophoresis devices with an efficient postcolumn reactor in planar quartz and glass chips. AB - Methods to fabricate planar capillary electrophoresis devices integrated with a postcolumn reactor in fused silica (quartz) and Pyrex glass are presented. Quartz is etched at ~1 MUm/min with a 2.1:1 width-to-depth ratio using a Cr/Au/Cr metal mask and concentrated HF/HNO(3). On-chip postcolumn reaction of o phthaldialdehyde (OPA) and amino acids gave theoretical plate numbers up to 83 000 and ~90 ms peak widths, corresponding to 14 plates/V and a 0.5 MUm theoretical plate height. The reactor geometry caused only a 10% degradation in efficiency. PMID- 21619341 TI - Electromigration injection from a small loop in capillary electrophoresis. AB - Capillary electrophoresis (CE) is now clearly the separation technique of the decade. One aspect that is still in need of improvement is concentration sensitivity, especially when CE is used in conjunction with on-column UV-visible absorptiometry, by far the most common practice. Electromigration injection (EI) is among the most prominent techniques in CE that serves to improve the attainable limits of detection; it is also the most convenient and simplest of all injection modes. Unfortunately, EI is affected both from sample to sample (the amount of an analyte introduced depends on sample conductance) and from analyte to analyte within a sample (the introduction is strongly biased on analyte electrical mobility). Previously we have shown the utility of small loops affixed at the tip of a capillary (Anal. Chem. 1995, 67, 3853-3860; 1996, 68, 1164-1168). The present paper shows that there are remarkable advantages to be gained from forming a film of the sample solution on a wire loop and using the loop itself as the high-voltage electrode to perform electromigration from a very small sample volume. The sample constituents can be essentially exhaustively electromigrated from this volume in less than 1 min, and the mobility induced bias is dramatically lowered. The observed experimental behavior agrees with theoretical models. PMID- 21619342 TI - Optical control of fusion of microparticles in solution and simultaneous spectrophotometric measurements. AB - A multibeam laser manipulation technique has been applied to noncontact and nondestructive optical fusion of two dye-containing microdroplets or that of a dye/water droplet and a porous silica gel particle in solution. Characteristics of the dye/particle(s) before and after coalescence have been followed simultaneously by microspectroscopy. Application of the multibeam laser manipulation-spectroscopy method in microanalytical chemistry is discussed. PMID- 21619343 TI - Detection of High Molecular Weight Narrow Polydisperse Polymers up to 1.5 Million Daltons by MALDI Mass Spectrometry. AB - The detection of very high molecular weight narrow polydisperse poly(styrene) samples by MALDI time-of-flight mass spectrometry is reported. It is shown that accurate molecular weight determinations of samples up to 1 million can be achieved very rapidly from the singly charged polymeric species. For a poly(styrene) with a molecular weight of approximately 1.5 million, signals corresponding to the multiply charged ions of the principal distribution are observed. The molecular weights obtained by MALDI are in good agreement with classical molecular weight determination techniques. all-trans-Retinoic acid was used as the organic matrix for the laser desorption procedure, and the samples were analyzed as their silver cation adducts. This work demonstrates that, with proper matrix selection and sample preparation, MALDI can be a very useful tool for high molecular weight polymer sample analysis. PMID- 21619344 TI - Gradient elution in capillary electrochromatography. AB - In analogy to pressure-driven gradient techniques in high-performance liquid chromatography, a system has been developed for delivering electroosmotically driven solvent gradients for capillary electrochromatography (CEC). Dynamic gradients with submicroliter per minute flow rates are generated by merging two electroosmotic flows that are regulated by computer-controlled voltages. These flows are delivered by two fused-silica capillary arms attached to a T-connector, where they mix and then flow into a capillary column that has been electrokinetically packed with 3-MUm reversed-phase particles. The inlet of one capillary arm is placed in a solution reservoir containing one mobile phase, and the inlet of the other is placed in a second reservoir containing a second mobile phase. Two independent computer-controlled, programmable, high-voltage power supplies (0-50 kV) [Formula: see text] one providing an increasing ramp and the other providing a decreasing ramp [Formula: see text] are used to apply variable high-voltage potentials to the mobile phase reservoirs to regulate the electroosmotic flow in each arm. The ratio of the electroosmotic flow rates between the two arms is changed with time according to the computer-controlled voltages to deliver the required gradient profile to the separation column. Experiments were performed to confirm the composition of the mobile phase during a gradient run and to determine the change of the composition in response to the programmed voltage profile. To demonstrate the performance of electroosmotically driven gradient elution in CEC, a mixture of 16 polycyclic aromatic hydrocarbons was separated in less than 90 min. This gradient technique is expected to be well suited for generating not only solvent gradients in CEC but also other types of gradients, such as pH and ionic strength gradients, in capillary electrokinetic separations and analyses. PMID- 21619345 TI - Synthesis and characterization of capillary columns coated with glycoside-bearing polymer. AB - Fused silica capillary columns were coated with a polymer obtained by radical polymerization of a novel acrylamido derivative, [(N-acryloylamino)ethoxy]ethyl beta-d-glucopyranose (AEG). This monomer, bearing a monosaccharide residue, was synthesized by direct coupling of [(N-acryloylamino)ethoxy]ethanol with glucose. The aim of this work was to demonstrate that weak, reversible interactions that take place between wall polymer coatings and protein analytes lead to poor reproducibility of transit times and capillary performance degradation. The pronounced hydrophilicity of the new poly(AEG) phase improves the reproducibility of transit times and the peak efficiency, as demonstrated by a complete series of tests carried out at various pH values. PMID- 21619346 TI - Condensation nucleation light scattering detection for capillary electrophoresis. AB - We describe two means for interfacing condensation nucleation light scattering detection to capillary electrophoresis (CE). With the first method, a fused silica capillary was used for the separation and the CE was grounded through a Nafion membrane that also connected the system to a microconcentric pneumatic nebulizer. Limits of detection (LODs) for underivatized amino acids were at the low microgram per milliliter level, and separation efficiencies were ~9 times lower than the optimum predicted for these species based on the injection plug width and axial dispersion by diffusion. LODs were limited by background nonvolatiles resulting from dissolution of fused silica at the high pHs used for the separations. An alternate system employed PEEK capillaries which acted as the separation capillary and also as the inner nebulizer capillary. In this case, the exit end of the capillary was coated with conductive paint which extended to the tip of the nebulizer, was in contact with the CE buffer, and was grounded to complete the CE circuit. Response was nonlinear and the separation efficiency of this system was somewhat lower than that for the Nafion membrane system. Response as peak heights for all of the amino acids and peptides studied was nearly identical on a mass basis. With this system, much lower background signals were obtained, and as a result, LODs for underivatized amino acids and peptides were below the 1 MUg/mL level, corresponding to less than 10 pg or less than 100 fmol injected. Both systems were fairly simple, effective means to generate aerosols with the low flows of CE and should be applicable to interfacing of other aerosol based detectors with CE. PMID- 21619347 TI - Fritless packed columns for capillary electrochromatography: separation of uncharged compounds on hydrophobic hydrogels. AB - A novel column is described that does not require frits to keep packing material within a capillary. A continuous bed is prepared in situ in aqueous solution by radical copolymerization of N-isopropylacrylamide and 2-acrylamido-2 methylpropanesulfonic acid (the resultant gel is denoted poly(AMPS-co-IPAAm). N,N'-Methylenebisacrylamide is used for cross-linking. On the application of an electrical field, electroosmotic flow (EOF) is developed in the bed along the capillary, where fluid propulsion would be otherwise difficult to achieve. The resultant EOF transports neutral compounds through the column without forcing the gel out of the capillary. Examination of the fluid motion in the continuous bed using a video microscope system and an image processor shows a relatively flat flow profile of EOF. The bed functions as the stationary phase for reversed-phase capillary electrochromatography (CEC). This new approach is an alternative to packed capillary columns which have been used previously in CEC. A high efficiency is obtained for a steroid which is separated on a 4.0% total monomer concentration (T), 10.0% degree of cross-linking (C), and 10.0% mole fraction of AMPS in the total monomer (S), poly(AMPS-co-IPAAm) column. A mixture of polyaromatic hydrocarbons is separated on a 6.9% T, 5.8% C, and 5.5% S poly(AMPS co-IPAAm) column. The capacity factor of benzo[a]pyrene increases from 0.63 to 1.91 as the acetonitrile content in a Tris-boric acid buffer is decreased from 45 to 30% (v/v). The run-to-run RSD of analyte migration time is less than 0.73%, and the day-to-day RSD is acceptable. Potential benefits of this approach are also mentioned. PMID- 21619348 TI - Acoustic flame detector for gas chromatography. AB - A novel gas chromatography detector is described that uses acoustic signals from a partly premixed hydrogen-air flame burning on top of a capillary. The device, referred to as the acoustic flame detector (AFD), is based on the measurement of the frequency of acoustic transients generated at the burner under a range of operating conditions. The presence of trace amounts of analyte in the flame was found to increase the frequency of these sonic bursts from the baseline level of ~100 Hz. The response of the AFD for n-dodecane, measured as the shift in frequency, was determined to be linear over ~3 orders of magnitude, with a minimum detectable level of about 1-5 ng C/s using the current system. The sensitivity correlates roughly with carbon content, except for certain organometallics (Sn, Mn), which gave substantially enhanced signals. Some tailing was observed but became serious only for particular types of organometallics. The noise of the system was predominantly of the 1/f type. The effects of flow conditions, burner geometry, and flame gas constituents were investigated. The oscillations could be followed by acoustic, visual, electrical, and optical means. The AFD mechanism is shown to involve oscillatory chemical kinetics, in which the flame front (the inner cone) temporarily enters a few millimeters into the capillary during each cycle, thereby creating the acoustic signal. PMID- 21619349 TI - Real-time fluorescence imaging of isotachophoretic preconcentration for capillary electrophoresis. AB - Real-time studies of the dynamic processes that take place during isotachophoresis (ITP) were performed. The experimental arrangement utilized was a real-time fluorescence imaging system based on a dye laser at 488 nm, pumped by a XeCl excimer laser. Fluorescence emitted from the migrating sample molecules was recorded by an image-intensified, thermoelectrically cooled CCD camera. The camera signals were processed by a computer and displayed on a screen in real time, allowing the ITP concentration to be monitored continuously. Real-time analysis provided additional information concerning ITP hardly obtainable using conventional detection systems or by theoretical calculations. Such experimental data can be evaluated and be compared with theoretical calculations. Information obtained by this detection technique is useful if ITP is to be used, for example as an on-line preconcentration technique in combination with capillary zone electrophoresis. PMID- 21619350 TI - Method for the accelerated measurement of electroosmosis in chemically modified tubes for capillary electrophoresis. AB - A traditional neutral marker approach is combined with fast pressure-driven mobilization to achieve a method that provides accelerated electroosmosis measurements in capillary electrophoresis systems. A marker injection is made prior to and following timed high-voltage application. The marker-sandwiched electrolyte plug is then pressure-driven out of the capillary tube under closely controlled conditions, while the signal is monitored in the usual way. Substantial reduction in measurement time is achieved by allowing the marker to travel only a fraction of the effective length (the injection to detection distance) over a shorter period of time, compared to the traditional method where voltage is applied until the neutral marker shows up at the detector window. One simple equation was derived to express the electroosmotic mobility of the electrolyte (MU(eo)) in terms of common operational variables. The method is particularly useful for low-electroosmosis measurements in wall-modified capillary tubes. The scope and limitations of the method are discussed. PMID- 21619351 TI - A surface ionization detector for gas chromatography: use of a supersonic free jet. AB - A new design of a gas chromatographic surface ionization detector (SID) based on hyperthermal positive surface ionization is described. There are two requirements: use of a supersonic free jet nozzle and the high-work-function surface of rhenium oxide. This detector, which is sensitive in response to all the organics, can be operated as an universal detector with much higher sensitivity toward some species having low ionization energy but with selectivity to a lesser degree than that of a conventional SID. The minimum detectable amount (at S/N = 3) of pyrene is around 4.4 * 10(-)(13) g/s, with linearity greater than 10(6), while that of toluene is around 10(-)(12) g/s. Other unique properties are (1) the ability to control the degree of selectivity through molecular kinetic energy, the surface, and its temperature and (2) a very short response time. PMID- 21619352 TI - Kinetic model of membrane extraction with a sorbent interface. AB - Membrane extraction with a sorbent interface (MESI) is an unique sample preparation alternative for trace organic analysis. The main features of MESI include its solvent-free nature, the rugged and simple design with no moving parts for long-term reliable performance, the fact that it is a single-step process which ensures good precision, its easy automation, and its feasibility for on-site operation. Among the available membrane extraction modules designed for the MESI system, the headspace configuration has continued to show its superior durability and versatility in membrane applications. The headspace membrane extraction configuration effectively eliminates the need for a sampling pump and flow metering and hence prevents the extraction system from plugging and greatly simplifies the extraction process. The module can be used for extraction of VOCs from gaseous, aqueous, or solid samples. A mathematical model has been developed for headspace membrane extraction of an aqueous sample, based on the assumption that the aqueous phase is perfectly stirred. The model is in good agreement with the experimental benzene extraction results obtained with an efficient agitation method such as high-speed magnetic stirring or sonication. The model has also been used to study the effects of various extraction parameters with respect to the sensitivity and response time of the MESI system. Sample agitation facilities analyte mass transport and hence improves both the system sensitivity and the response time. The sensitivity of the extraction method also increases with an increase of the extraction temperature. PMID- 21619353 TI - Immobilization of crown ether carboxylic acids on silica gel and their use in column concentration of alkali metal cations from dilute aqueous solutions. AB - Eight crown ethers with pendent carboxylic acid groups are immobilized on silica gel and utilized for column concentration of alkali metal cations from dilute aqueous solutions. The column concentration selectivity and efficiency are found to be strongly influenced by (1) the cavity size of the crown ether unit, (2) conformational positioning of the proton-ionizable side arm with respect to the crown ether cavity, and (3) capping of residual silanol surface groups with trimethylsilyl functions. By use of a chromatographic stripping technique, selective column concentration of Na(+), K(+), (Rb(+) and Cs(+)), and Cs(+) by different functionalized silica gels has been achieved. PMID- 21619354 TI - Reversed phase liquid chromatography of organic hydrocarbons with water as the mobile phase. AB - Reversed phase high-performance liquid chromatography (RP-HPLC) is demonstrated for hydrophobic analytes such as aromatic hydrocarbons using only water as the mobile phase. Achievement of reasonable capacity factors for these types of compounds without the need for toxic and costly organic modifiers in the mobile phase is accomplished by substantially decreasing the phase volume ratio of stationary phase relative to the mobile phase volume and by increasing the polarity of the stationary phase relative to stationary phase materials commonly used for RP-HPLC. Applying a stationary phase of trifluoropropylsiloxane, which is a common gas chromatographic stationary phase material, to nonporous glass microspheres yields a stationary phase with a phase volume ratio reduced by about 2 orders of magnitude as compared to common liquid chromatographic packing materials. As a result, a separation was obtained for hydrophobic organic analytes such as benzene, toluene, ethylbenzene, and isopropylbenzene using a water mobile phase at ambient temperature. A separation of sodium benzoate, benzaldehyde, benzene, and butyrophenone is shown in less than 3 min using a water mobile phase and UV/visible absorbance detection. Additionally, the separation of the ionic surfactant species octyl sulfate and dodecyl sulfate in water in less than 3 min, using unsuppressed conductivity detection, is achieved with a separation mechanism based on interactions with the hydrophobic portion of the surfactant. A water mobile phase offers many potential advantages over traditional mixed aqueous/organic solvent systems. In addition to saving on the cost and expense of buying and disposing of toxic solvents and waste, there is less exposure of the operator to potentially harmful solvents. Increased consistency in reproducing retention times can be expected, since there will not be any variability in solvent strength due to slight variations in mobile phase composition. A water mobile phase produces an environment that should provide an inherent advantage of increased signal-to-noise ratio for detection. Additionally, excellent predictions of the octanol/water partitioning coefficient and aqueous solubility for hydrophobic analytes are obtained from a single measurement of the capacity factor in the water mobile phase. PMID- 21619355 TI - Retention mechanism of weak polar solutes in reversed phase liquid chromatography. AB - The retention mechanism of a weak polar solute, a series of 10 benzodiazepines in reversed phase liquid chromatography, was investigated over a wide range of mobile phase compositions. The values of enthalpy (DeltaH degrees ) and entropy (DeltaS degrees ) of transfer from the mobile to the stationary phases were determined. The method studied each factor (water fraction Phi in the acetonitrile (ACN)/water mixture and column temperature) controlling the retention mechanism. The changes in DeltaH degrees and DeltaS degrees as a function of the water fraction Phi in the ACN/water mixture were examined. These variations are explained using the organization of organic modifier (ACN) in clusters in the ACN/water mixture. A change in the retention mechanism thus indicated when the ACN/water mixture was used instead of the hydrogen-bonded mobile phase such as CH(3)OH/water. Enthalpy-entropy compensation revealed that the retention mechanism was independent of the water fraction Phi but showed that differences between the molecular structures of the benzodiazepines contributed more significantly to changes in the retention process in the CH(3)OH/water mixture than in the ACN/water mixture. PMID- 21619356 TI - A Comparison of Cryofocusing Injectors for Gas Sampling and Analysis in Fast GC. AB - The performance of two cryofocusing injectors for fast gas chromatography has been studied. The first system traps analytes onto bare metal tubes and rapidly vaporizes them upon ballistically heating the tube using a capacitance discharge. The second is a microloop injector in which analytes are cryotrapped onto short lengths of narrow-bore fused silica tubing with various coatings. The ballistically heated injector is capable of sampling and injecting compounds from air faster than the microloop system, because the metal tube can be heated and cooled more rapidly. Both systems are capable of cryotrapping compounds as volatile as butane at -90 degrees C, and the microloop system can trap ethane when a section of a porous layer open-tubular (PLOT) column is used as the sample loop. In addition, the microloop injector can be used without cryointegration to analyze compounds regardless of their volatility, as long as they are present in the samples at detectable concentrations. Because the ballistically heated injector is flushed prior to injection, it can introduce only compounds that are adsorbed onto its metal trap. Comparison of chromatograms obtained using the two injectors show similar chromatographic resolution. Both traps are susceptible to freezing during the cryotrapping step, but the use of an inline Nafion dryer allows air saturated with water vapor to be sampled using both systems for 3 min without plugging the trap. Thermal decomposition during the injection step can occur for labile species in the ballistically heated trap, but even the highly unstable compound ethyl diazoacetate may be injected without breakdown in the microloop system. PMID- 21619357 TI - Lateral diffusion of molecules partitioned into silica-bound alkyl chains: influence of chain length and bonding density. AB - Lateral diffusion of a hydrophobic fluorescent molecule partitioned into monomeric alkyl chains bound to a planar silica substrate was measured as a function of chain density and chain length. Measurement of fluorescence recovery after patterned photobleaching was used to observe the diffusional relaxation of a concentration profile of probe molecules over distances of micrometers. The diffusion rate of the probe molecule partitioned into C-18 chains decreased with decreasing chain coverage. As the chain length was reduced from C-18 to C-8 and C 4, the rate of diffusion also decreased. These results, when combined with results from a previous study of the effect of overlaying solvent on diffusion rate (Hansen, R. L.; Harris, J. M. Anal. Chem. 1995, 67, 492-498), are consistent with a domain model for long-range transport of partitioned molecules through the bound ligands. Fluorescence recovery experiments in which diffusion is monitored over a distance of micrometers offer a unique means to probe long-range structure of surface-immobilized alkyl chains. PMID- 21619358 TI - Characterization of Nonbonded Poly(ethylene oxide) Coating for Capillary Electrophoresis via Continuous Monitoring of Electroosmotic Flow. AB - We examined changes in a poly(ethylene oxide) (PEO) coating by continuously monitoring the electroosmotic flow (EOF) in a fused-silica capillary during electrophoresis. An imaging CCD camera was used to follow the motion of a fluorescent neutral marker zone along the length of the capillary. The PEO coating was shown to reduce the velocity of EOF by more than 1 order of magnitude compared to a bare capillary at pH 7.0. However, it did not reduce EOF efficiently at pH 8.2. The coating protocol was important, especially at an intermediate pH of 7.7. Capillary reconditioning with an acidified solution of PEO was necessary in order to create a stable and efficient coating. In all cases we observed a gradual increase of EOF during extended runs, suggesting that the coating is slowly being degraded. The increase of pH in the cathodic (detection end) buffer reservoir beyond pH ~8.0, e.g., as a result of electrolysis, had a large impact on the stability of the coating. This phenomenon may be used for the efficient and fast regeneration of the column surface and provides a simpler and more reliable alternative to pressure flushing of the capillary. PMID- 21619359 TI - A renewable-reagent fiber-optic sensor for measurement of high acidities. AB - A renewable-reagent fiber-optic HNO(3) sensor was developed for HNO(3) measurement in the 0.1-10.0 M range. The HNO(3) sensor employs a tubular Nafion cation-exchange membrane to extract acid species from an external HNO(3) sample into an internal flowing reagent solution. In high-concentration HNO(3) samples, incomplete HNO(3) dissociation results in a significant concentration of neutral HNO(3) species in addition to protons. As both neutrals and protons are potentially membrane-permeable species, various reagent compositions were tested to examine the contributions of both acid transfer mechanisms. Continuous reagent flow limited internal acid accumulation and transferred reagent to the sensor optical detection cell. All reagent compositions included cresol red as a colorimetric indicator, which was measured within the sensor detection cell. Careful fiber-optic alignment provided sufficient light throughput in a backscatter illumination mode to allow use of a photodiode array detector for visible spectral acquisition. The use of Ca(2+) as a reagent countercation produced notable reductions in HNO(3) sensor response to interferent cations and temperature changes. Sensor measurement of HNO(3) samples in the tested concentration range produced average relative standard deviations of less than 0.4%. Control over reagent flow rate should allow for extension of the HNO(3) sensor measurement range to 16.0 M HNO(3). PMID- 21619360 TI - Appearance of discontinuities in spectra transformed by the piecewise direct instrument standardization procedure. AB - Several years ago, we noted that spectra transformed by the piecewise direct standardization (PDS) method may contain discontinuities. Having noticed that the problem was a recurring one, we studied it and recently diagnosed its source. Our investigations suggest that this problem also occurs in applications of window factor analysis, evolving factor analysis, and any other procedure that uses piecewise principal component models. In this work, we report the source of the problem and illustrate it with one example. A procedure is presented for eliminating the problem that is effective in PDS pattern recognition applications. Further work is needed to develop modified algorithms suitable for calibration applications. PMID- 21619361 TI - Linamarin sensors: interference-based sensing of linamarin using linamarase and peroxidase. AB - An interference-based linamarin sensor is developed. Horseradish peroxidase (HRP) is adsorbed on a pyrolytic graphite (PG) electrode, and then linamarase from cassava is cross-linked with glutaraldehyde on the electrode surface. The prepared bienzyme electrode is poised at -300 mV vs Ag/AgCl for 40 s to reduce dissolved O(2) to H(2)O(2) at the PG surface. The potential is then stepped to 0 mV, at which point the accumulated H(2)O(2) is reduced, though the O(2) reduction does not proceed. Since the H(2)O(2) reduction is catalyzed by HRP, the transient cathodic current is inhibited by cyanide, which is liberated from linamarin by linamarase. Therefore, the transient current is a function of the linamarin concentration. This sensor responds to 1 * 10(-5)-5 * 10(-3) M linamarin and can estimate a linamarin concentration of a cassava extract. PMID- 21619362 TI - Interdigitated array electrode as an alternative to the rotated ring-disk electrode for determination of the reaction products of dioxygen reduction. AB - Electrochemical reduction of dioxygen in aqueous media can proceed to water, hydrogen peroxide, or a mixture of the two. The production of hydrogen peroxide, classically established with the rotated ring-disk electrode, can also be quantitatively assessed at interdigitated array (IDA) electrodes, where dioxygen is reduced at the set of microband generator electrodes and any H(2)O(2) produced is detected by its oxidation (back to O(2)) at the interdigitated set of microband collector electrodes. The sensitivity of the IDA for H(2)O(2) detection is higher owing to its more complete collection, and to the ensuing regeneration of O(2), which leads to an amplification of the generator currents. The production of H(2)O(2) is thus reflected both in the ratio of collector and generator electrode currents [the collection efficiency, coll(tau)] and in the ratio of the generator current with the collector potential on to that with it off [amplification factor, ampl(tau)]. The necessary theory for interpretation of the fraction epsilon of H(2)O(2) produced per dioxygen reduced is presented, based on conformal mapping techniques. Explicit equations are derived for epsilon at long times that are independent of the IDA dimensions and that can be used with any two-product electrochemical reaction analogous to the dioxygen reduction. Experimental data are presented for dioxygen reduction in acidic and basic media to illustrate application of the theory. PMID- 21619363 TI - Detection of Partial Denaturation in AT-Rich DNA Fragments by Ion-Pair Reversed Phase Chromatography. AB - Temperature-dependent denaturation of DNA restriction fragments from the pBR322 plasmid ranging in length from 46 to 910 base pairs was detected by ion-pair reversed-phase high-performance liquid chromatography using columns packed with alkylated nonporous poly(styrene/divinylbenzene) particles. The presence of acetonitrile in the mobile phase was found to decrease the melting temperatures of DNA fragments by 1.5-2 degrees C/% of acetonitrile in the eluent. Small fragments (<120 bp) were completely denatured between 53.6 and 63.5 degrees C, depending on their total GC content. Whereas retention times of completely helical DNA fragments increased gradually with increasing temperature, partial denaturation of larger DNA fragments (>150 bp) was found to reduce retention at temperatures above 53.6 degrees C. Therefore, micropreparative fractionation and rechromatography, together with DNA restriction analysis, were applied to identify the correct elution order of completely helical and partially denatured fragments. Inspection of the DNA sequences of partially denatured fragments revealed domains with repeating AT base pairs. Positions of partial denaturation within the pBR322 plasmid detected by chromatographic analysis were in good agreement with partial denaturation maps obtained by electron microscopy desrcibed in the literature. PMID- 21619364 TI - Spectroscopic imaging of laser-induced plasma. AB - Spectroscopic imaging provides 2D images with full spectral resolution at each pixel. Thus, chemical imaging of an object, as well as other useful information, can be obtained. An imaging spectroscopy method in the visible range is presented and applied to laser plasma. This is a powerful research tool with numerous possible applications. This study is focused on spectroscopic imaging of laser produced plasmas, and such spectral images (full spectrum at each pixel) are presented for the first time. Detailed information on optical and geometrical effects are obtained, and an insight to the optimization of the laser plasma spectroscopy method is achieved. The size and the spatial shape of the plasma, which can be used for matrix effect compensation, are measured. Similarity maps and classification maps of laser-induced plasma are obtained for the first time. These maps are used for allocation of chemical components in the plasma. The signal to noise ratio maps of the spectra obtained from laser-induced plasmas are provided. These surfaces possess a clear maximum, indicating that there is a preferred site in the plasma, where the emitted light provides the best signal to noise ratio. The performance of the current method is limited by the lack of temporal resolution, although it can be extended by a proper temporal gating. PMID- 21619365 TI - Ultratrace determination of selected lanthanides by luminescence enhancement. AB - The luminescence intensity of trivalent lanthanides, especially terbium(III) and europium(III), is shown to be enhanced by coordination with the ligand 2,6 pyridinedicarboxylic acid (DPA). Further enhancement can be obtained by forming a columinescent complex aggregate with ions such as gadolinium(III), terbium(III), lanthanum(III), or yttrium(III), where the Y(III) complex shows the greatest enhancement. A surfactant, sodium dodecyl sulfate (SDS), can be added to the solution to enhance the luminescence intensity of many lanthanide-ligand complexes by segregating the complex from quenchers. In this study, SDS has little effect on the limit of detection, but it does act to extend the linear dynamic range to include higher concentrations. The combination of columinescent complex with surfactant resulted in decreased luminescence intensity coupled with an increase in the background due to the light scattered by the surfactant micelles. A mechanism for the enhancement of the lanthanide luminescent intensity by energy transfer has been described by Xu et al. This study differs in that a lanthanide ion excitation band represents the most efficient excitation path and not a ligand excitation band. The complexes examined in this study have advantages over those used in prior studies since they do not require surfactants to achieve low limits of detection (100 parts per quadrillion, ppq, for Eu(DPA)(3)(3)(-) and 60 ppq for Tb(DPA)(3)(3)(-)), and they exhibit longer linear dynamic ranges (from 4 to 6 decades) than other columinescent systems. PMID- 21619366 TI - Determination of an iron suspension in water by laser-induced breakdown spectroscopy with two sequential laser pulses. AB - We have applied laser-induced breakdown spectroscopy to quantitative analysis of colloidal and particulate iron in water. A coaxial sample flow apparatus developed in our previous work, which allowed us to control the atmosphere of laser-induced plasma, was used. Using sequential laser pulses from two Q-switched Nd:YAG lasers as excitation sources, the FeO(OH) concentration in the tens of ppb range was determined with an optimum interval between two laser pulses and an optimum delay time of a detector gate from the second pulse. The detection limit of Fe decreased substantially using two sequential laser pulse excitations: the 0.6 ppm limit of single pulse excitation to 16 ppb with sequential pulse excitation. The effects of the second laser pulse on the plasma emission were studied. The concentration of iron in fine particles in boiler water sampled from a commercially operated thermal power plant has been determined successfully by this method. The results show the capability of laser-induced breakdown spectroscopy in determining suspended colloidal and particulate impurities in a simple and quick way. PMID- 21619367 TI - Transfer of Near-Infrared Multivariate Calibrations without Standards. AB - A novel approach to the transfer of multivariate calibration is proposed. This method is based on the finite impulse response (FIR) filtering of a set of spectra to be transferred, using a spectrum on the target instrument to direct the filtering process. Often, the target spectrum is the mean of a calibration set. The method is compared against direct transfer and piecewise direct transfer on near-infrared reflectance spectra in two representative data sets. Results from these studies suggest that FIR transfer compares favorably with piecewise direct transfer in terms of accuracy and precision of the match of transferred spectra to the predictive calibration models developed on the target instrument. Unlike piecewise direct transfer, FIR transfer requires no measurement of standard samples on both the source and target spectrometers. Details and current limitations of the FIR transfer method are presented. PMID- 21619368 TI - Determination of tetraethyllead and inorganic lead in water by solid phase microextraction/gas chromatography. AB - A new method for the determination of tetraethyllead (TEL) and ionic lead in water by SPME has been developed. TEL is extracted from the headspace over the sample. Inorganic lead is first derivatized with sodium tetraethylborate to form TEL, which is extracted in the same way as pure TEL samples. The analytical procedure was optimized with respect to pH, amount of derivatizing reagent added, stirring conditions, and extraction time. The detection limit obtained for TEL was found to be 100 ppt when using FID and 5 ppt when using ion trap MS (ITMS). The detection limit for Pb(2+), limited by the nonzero blank, was found to be 200 ppt. Linear calibration curves were obtained for both analytes when FID was used for detection. For lead they spanned over 4 orders of magnitude. ITMS offered excellent sensitivity and selectivity, but the calibration curves were nonlinear when the m/z = 295 ion was used for quantitation. The method has been verified on spiked tap water samples. An excellent agreement was found between the results obtained for standard solutions prepared using NANOpure water and spiked tap water samples. PMID- 21619369 TI - Stereospecific Derivatization of Amphetamines, Phenol Alkylamines, and Hydroxyamines and Quantification of the Enantiomers by Capillary GC/MS. AB - The enantiomers of amphetamines, phenol alkylamines, and hydroxyamines are separated by using alpha-methoxy-alpha-(trifluoromethyl)phenylacetyl chloride as the chiral derivatizing agent for amino groups. Prior to N-acylation, amine salts are converted into the free bases and hydroxyl groups into O-silyl ethers by reaction with N-methyl-N-silylamides. N-Methyl-N (trimethylsilyl)trifluoroacetamide, N-methyl-N-(triethylsilyl)trifluoroacetamide, or N-methyl-N-(tert-butyldimethylsilyl)trifluoroacetamide was used to protect the hydroxyl groups by TMS, TES, or the tBDMS groups. All these N-methyl-N silylamides were able to convert amino salts to the free bases. The reaction is selective and rapid, and the diastereomeric derivatives are well separated by capillary gas-liquid chromatography. This procedure is suitable for simultaneous determination by gas chromatography/mass spectrometry with selected-ion monitoring and is also applicable to quantification of the compounds in a biological matrix. PMID- 21619370 TI - Quantitative analysis and structure determination of styrene/methyl methacrylate copolymers by pyrolysis gas chromatography. AB - A pyrolysis gas chromatography (Py-GC) method has been developed to study the composition and microstructure of styrene/methyl methacrylate (STY/MMA) copolymers. The composition was quantified by Py-GC using monomer peak intensity. Because of the poor stability of methyl methacrylate oligomers, neither MMA dimer nor MMA trimers were detected under normal pyrolysis conditions. The number average sequence length for STY was determined by pure and hybrid trimer peak intensities. The number-average sequence length for MMA was determined by using formulas that incorporate composition and the number-average sequence length of STY. This method is a new approach for the investigation of the microstructure of those copolymers that do not produce dimer and trimer peaks upon pyrolysis. PMID- 21619371 TI - Comparison of extraction techniques, including supercritical fluid, high-pressure solvent, and soxhlet, for organophosphorus hydraulic fluids from soil. AB - The efficiencies of three extraction techniques for removal of nonpesticidal organophosphates from soil were determined. Traditional Soxhlet extraction was compared to supercritical fluid extraction (SFE) and a low solvent volume flow through technique referred to here as high-pressure solvent extraction (HPSE). SFE, optimized by varying parameters of temperature, pressure, and methanol polarity modifier, showed at least 90% efficiency in the extraction of OPs from both spiked and native soils. HPSE experiments showed efficient and consistent recoveries over a range of temperatures up to 200 degrees C and pressures up to 170 atm. Recovery of TCP from spiked soils with HPSE depends on the system variables of temperature and pressure, which dictate density and flow rate. HPSE provided extraction efficiencies comparable to those obtained with Soxhlet extraction and SFE but with substantial savings of time and cost. PMID- 21619372 TI - High-pressure stopped-flow spectrometer for kinetic studies of fast reactions by absorbance and fluorescence detection. AB - The development of a stopped-flow instrument that operates over a temperature range of -40 to +100 degrees C and up to 200 MPa is described. The system has been designed so that measurements can be performed in absorbance and fluorescence modes simultaneously, without dismantling the unit. It can easily be combined with an optical system of a conventional ambient pressure setup by using light guides. Optimum optical performance and a wide operating wavelength range (220-850 nm) are achieved as the light is not passing through the pressurizing fluid. A special design for the pistons has been developed; thus, the apparatus has proven to be leak-free, even under extreme conditions (high pressure, low temperature, various solvents). The dead time of the system is found to be less than 2 ms at 298 K and is pressure independent up to 200 MPa. We examined the kinetics for the formation of the Mg(2+)-8-hydroxyquinoline chelate in aqueous solutions at pH 8.0 in order to develop a convenient alternative test method for high-pressure stopped-flow spectrometers with absorption and fluorescence detection. PMID- 21619373 TI - Russian doll type cryogenic traps: improved design and isotope separation effects. AB - An improved cryogenic trap for removing micromole quantities of condensables from gas mixtures is described. It is based on the so-called Russian doll design, with which extraordinary trapping efficiencies at flow rates of up to 10 L/min are obtained. The active element consists of one or more nested glass fiber thimbles. Despite the large fiber area, quantitative retrieval of condensed CO(2) is obtained. The new design is demountable and incorporates a heat exchange section; trapping efficiency depends little on the level of coolant, as has been tested for CO(2). When fractionation is induced by incomplete trapping, a small isotopic enrichment in (13)C occurs. It also has been discovered that when He is used as carrier gas, a gas chromatographic effect for CO occurs in Russian doll traps, accompanied by a large isotope separation, in which the elution sequence corresponds to the vapor pressure ratios of the isotopomers. PMID- 21619374 TI - A simple method for insulating carbon-fiber microelectrodes using anodic electrophoretic deposition of paint. AB - We describe a simple method for the insulation of carbon-fiber microelectrodes (CFMEs). Using the technique of anodic electrophoretic deposition of paint (EDP), we deposited thin and uniform films of electrodeposition paint onto 10-MUm diameter carbon fibers. The polymer films were then heat cured, leading to an electrically insulating coat. The insulated carbon fibers were transected perpendicular to their axes to expose a 10-MUm carbon disk and characterized using conventional electrochemical methods and scanning electron microscopy. As expected, cyclic voltammograms measured with electropainted CFMEs in solutions containing ferricyanide displayed a sigmoidal response without hysteresis. The insulating films had a low dielectric constant, resulting in low capacitance. In addition, the film thickness could be controlled simply by varying such deposition parameters as the applied voltage and the duration of treatment. Electrical noise of the transected fibers was determined principally by the cut surface of the fiber, not by the exposed insulated surfaces. Compared to electropolymerization of phenol compounds, another electrochemical method for insulating CFMEs, insulation by anodic EDP has the very significant advantages of greater simplicity, much shorter processing time, and much lower toxicity of the reagents involved, important considerations for those who work with the procedure on a daily basis. Furthermore, electrodeposition paints are commercially available and have long shelf lives. PMID- 21619375 TI - Miniaturized ultraviolet ozonesonde for atmospheric measurements. AB - The design and test results of a simple single-beam ultraviolet photometer for ozone measurements are presented. The instrument has several features that distinguish it from former UV absorption instruments, all of which contribute to a smaller size and much lower power consumption while retaining the advantages of the UV measurement technique. A novel airflow system and advanced electronics are among the most important changes from earlier reported systems. The instrument fits into a package 10 cm in diameter by 60 cm long and weighs under 1.5 kg. Hence, it is well-suited for lightweight airborne applications using kites and balloons, as well as other portable measurement needs. Independent measurements of ozone are made every 4 s, with a sensitivity of 0.3 ppbv ozone (limit of detection for S/N = 3) and a precision of +/-2%. PMID- 21619376 TI - Improved scanning range for coherent anti-stokes Raman spectroscopy using a tunable optical parametric oscillator. AB - Coherent anti-Stokes Raman spectroscopy (CARS) is a well-known form of nonlinear spectroscopy that has been used for a wide range of specialized quantitative applications. From an analytical chemist's point of view, however, conventional CARS is impractical as a tool for qualitative and quantitative analyses because the scan range is too short to produce complete vibrational spectra. This paper introduces a new technique, synchronously scanned optical parametric oscillator (OPO) CARS, that improves the potential for using nonlinear spectroscopy as an analytical technique in both gas- and condensed-phase samples. First, it uses a broadly tunable OPO to increase the scan range. Second, phase matching problems that limit scans in condensed-phase CARS are reduced by using both the signal and the idler beams in a synchronous scanning manner. Finally, this synchronous scanning method generates an output signal that remains fixed at a single wavelength (single-wavelength detection). Advantages of single-wavelength detection include reduction of stray light, simplicity, and elimination of the need for wavelength calibration of the detection optics. Results are presented on neat and mixed samples in gas and condensed phases. PMID- 21619377 TI - Valved sampling cell for membrane introduction mass spectrometry. AB - Membrane extractors comprising a membrane house inside of a valve have been developed to separate compounds of interest from a sample matrix and introduce these compounds into a mass spectrometer. Experimental control over parameters that affect permeability or that may damage the membrane, such as the membrane temperature, is provided with the valve. The valve was tested for response and response times with the valve separated from the mass spectrometer by various interface tube lengths. Data for steady state response measurements showed no significant change with the valve at different distances from the ion source. Polar compounds show a strong response time dependency on the interface tube length. This adsorption phenomenon is minimized by simply heating the interface tube. Other factors affecting the performance of the device are discussed. PMID- 21619378 TI - The role of proprotein convertase subtilisin/kexin type 9 in hyperlipidemia: focus on therapeutic implications. AB - Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a key regulator of circulating levels of low-density lipoprotein (LDL) particles. PCSK9 acts mainly by enhancing degradation of the LDL receptor in the liver. Several gain-of function and loss-of-function mutations in the PCSK9 gene have been identified and linked to hypercholesterolemia and hypocholesterolemia, respectively. Since the loss-of-function mutations in humans are associated with protection against coronary heart disease, and with no apparent deleterious effects, PCSK9 inhibition is becoming attractive as a new strategy for lowering LDL cholesterol (LDL-C) levels, particularly in combination with statins. Candidate patient populations for PCSK9 inhibition include those with familial hypercholesterolemia, patients at high risk of cardiovascular disease not controlled by statin therapy, and patients with poor tolerance or total intolerance to statin therapy. PCSK9 inhibition represents a very promising target for reducing LDL-C levels and decreasing the risk of atherosclerotic cardiovascular diseases, but human clinical trials will be crucial to assess the potency and safety of PCSK9 inhibitors. PMID- 21619379 TI - Chronic heart failure: current evidence, challenges to therapy, and future directions. AB - Heart failure (HF) is a complex syndrome characterized by the inability of the heart to maintain a normal cardiac output without elevated intracardiac filling pressures, resulting in signs of pulmonary and peripheral edema and symptoms of dyspnea and fatigue. Central to the management of HF is a multifaceted pharmacological intervention to abate the harmful counter-regulatory effects of neurohormonal activation and avid salt and water retention. Whereas up to 40 years ago HF was managed with diuretics and leaf of digitalis, the cornerstones of therapy for HF patients with systolic dysfunction now include ACE inhibitors or angiotensin II type 1 receptor antagonists (angiotensin receptor blockers), beta-adrenoceptor antagonists (beta-blockers), and aldosterone antagonists, which have significantly improved survival. However, with the increasing number of beneficial therapies, there are challenges to implementing all of them. Specific cardiomyopathies also merit specific considerations with respect to treatment, and - unfortunately - there is no therapy for HF with preserved left ventricular ejection fraction that has been shown to improve survival. Although mortality has improved in HF, the biggest challenge to treatment lies in addressing the morbidity of this disease, which is now the most common reason for hospital admission in our aged population. As such, there are many therapies that may serve to improve the quality of life of HF patients. Future HF treatment regimens may include direct cellular therapy via hormone and cytokine signaling or cardiac regeneration through growth factors or cell therapy. PMID- 21619380 TI - Prognostic utility of serum potassium in chronic digoxin toxicity: a case-control study. AB - OBJECTIVE: In contrast to patients with acute digoxin overdose, the prognostic utility of the serum potassium concentration for patients with chronic digoxin toxicity is unclear. In such patients, we aimed to evaluate the relationship between pre-treatment serum potassium and survival. METHODS: This was a case control study at an urban Poison Control Center affiliated with a large urban medical center. We compared the serum potassium concentration between patients with chronic digoxin toxicity resulting in fatality (cases) over a 7-year period (2000-2006) versus survivors (controls) over a 1-year period (2007-2008). RESULTS: During the study period, there were 13 fatalities (cases) and 13 survivors (controls), of whom seven cases and five controls received appropriately dosed digoxin-specific antibody Fab fragments (Fab). There were no statistically significant differences between cases and controls with respect to serum digoxin concentration, creatinine, age, or sex. Serum potassium elevation pre-Fab was significantly associated with fatality both in mean difference (p < 0.03) and using a dichotomous cutoff of 5.0 mEq/L (p < 0.001), which performed with 92% sensitivity (95% CI 67, 99). In 86% of deaths despite appropriate Fab administration, the clinical presentation included the combination of bradycardia plus hyperkalemia. CONCLUSION: In these patients with chronic digoxin toxicity, elevated serum potassium was associated with fatality. The combination of bradycardia and hyperkalemia strongly predicted fatality even in cases with appropriate Fab administration. PMID- 21619381 TI - Niacin extended-release therapy in phase III clinical trials is associated with relatively low rates of drug discontinuation due to flushing and treatment related adverse events: a pooled analysis. AB - BACKGROUND AND OBJECTIVE: Niacin is a highly effective agent for increasing low high-density lipoprotein cholesterol (HDL-C) levels. It also has beneficial effects on key pro-atherogenic lipoprotein parameters. However, the side effect of flushing can challenge patient adherence to treatment. In this study, we pooled safety data from available trials of at least 16 weeks' duration to evaluate the impact of flushing on patient adherence to niacin extended-release (NER) therapy. METHODS: Data were pooled from eight NER studies (administered as NER with a maximum dosage of 1000, 1500, and 2000 mg/day, either as monotherapy or in combination with simvastatin 20 or 40 mg/day [NER/S], or lovastatin 20 or 40 mg/day [NER/L]) to evaluate rates of study discontinuation due to flushing or any treatment-related adverse events. RESULTS: While 66.6% of patients experienced flushing, only 5.2% of patients discontinued treatment due to flushing. Of the total number of patients treated with NER (n = 307), NER/S (n = 912), or NER/L (n = 928), 34 (11%), 105 (11%), and 127 (14%) patients discontinued due to any treatment-related adverse event, respectively, while 14 (5%), 43 (5%), and 55 (6%) discontinued due to flushing. Discontinuation for flushing did not differ with regard to maximum dose, or to the presence or type of statin combined with NER. CONCLUSION: Although flushing was common with NER treatment, discontinuation due to flushing occurred in only 5-6% of patients in this pooled analysis. This could be due to several factors, including the fact that all patients in the NER trials were educated about flushing and its management. Translation of methodology employed in these trials into clinical practice may improve long-term adherence to NER therapy, which would enhance the therapeutic benefit of NER for reducing cardiovascular risk. PMID- 21619382 TI - ACE inhibition modulates endothelial apoptosis and renewal via endothelial progenitor cells in patients with acute coronary syndromes. AB - BACKGROUND: The equilibrium between endothelial apoptosis and endothelial renewal is altered in acute coronary syndromes and may be related to differences in the beneficial effects of angiotensin-converting enzyme inhibitors and angiotensin II receptor antagonists (angiotensin receptor blockers). METHODS: We evaluated the effect of treatment on endothelial function in post-myocardial infarction (MI) patients treated with perindopril (group 2, n = 16) or valsartan (group 3, n = 17) at baseline and after 7, 15, and 30 days and in normal controls (group 1, n = 20). Endothelial apoptosis was determined by cultivating serum samples in vitro with human umbilical vein endothelial cells (HUVECs), while endothelial renewal was assessed by mobilization of CD34+ bone marrow cells. RESULTS: At baseline, post-MI patients had significantly elevated rates of apoptosis (16.6 +/- 5.0% and 16.5 +/- 8.4% in groups 2 and 3, respectively [both p = 0.01] vs 1.6 +/- 0.7% in group 1), which declined in group 2 (10.5 +/- 4.4% at 30 days, p = 0.04), but not in group 3. Similar results and trends were found for the Bax/Bcl-2 ratio. CD34+ mobilization was significantly increased in group 2 (3.0 +/- 1.0 at baseline to 6.2 +/- 1.6 at 15 days, p = 0.03), whereas in group 3 CD34+ mobilization did not change significantly. The findings in group 2 were accompanied by an increase in vascular endothelial growth factor at 15 days, and a reduction in tumor necrosis factor-alpha and its soluble receptors, versus no change in group 3. Similar findings were observed for angiotensin II and bradykinin. CONCLUSION: Our results indicate that perindopril, but not valsartan, reduces the proapoptotic effect of serum on the endothelium and increases endothelial renewal in patients with acute coronary syndromes. PMID- 21619383 TI - QT response after a test dose and during maintenance therapy with AZD1305 in patients with atrial fibrillation: a double-blind, randomized, placebo-controlled trial. AB - BACKGROUND AND OBJECTIVE: AZD1305 is an investigational antiarrhythmic agent that prolongs refractoriness through combined potassium and sodium channel inhibition. This study aimed to explore the utility of a test dose in predicting QT interval corrected according to Fridericia's formula (QTcF) during subsequent maintenance treatment with AZD1305. METHODS: This was a randomized, double-blind, parallel group, placebo-controlled trial carried out at multiple hospital cardiac facilities in Denmark, Norway, Poland, Slovakia, and Sweden. Patients with documented atrial fibrillation (AF) but currently in stable sinus rhythm for >=2 hours and <=90 days were eligible for inclusion. Patients were randomized in a 1 : 1 : 1 ratio to receive AZD1305 extended-release or matching placebo tablets as follows: group A - test dose 250 mg, evening dose 125 mg on day 1, maintenance dose 125 mg twice daily; group B - test dose 500 mg, placebo evening dose, maintenance dose 125 mg twice daily; placebo group - placebo test and maintenance dose. Maintenance dosing was for 9 days. QTcF >550 ms at any time during the in patient phase or >500 ms after discharge (day 4) were predefined study drug discontinuation criteria. The main outcome measure was the relationship between QTcF following the test dose and during maintenance treatment. RESULTS: Sixty five patients were randomized (n = 21, 22, and 22 in group A, group B, and the placebo group, respectively). AZD1305 dose-dependently increased QTcF. There was a positive, linear correlation between the change in QTcF during the first 6 hours after the test dose and during the maintenance phase. Three patients, all from group B, discontinued treatment on day 1 due to QTcF >550 ms. All other patients completed the study without events related to QT prolongation. There was a trend for reduced AF recurrence with AZD1305 compared with placebo. CONCLUSION: In this exploratory study a test dose predicted the QT response during maintenance treatment with AZD1305 and may thus be employed in further studies. [ClinicalTrials.gov Identifier: NCT00643448]. PMID- 21619384 TI - Neointimal hyperplasia inhibition effect of angiotensin II type 1 receptor blockers in patients after coronary stent implantation: a meta-analysis. AB - BACKGROUND AND OBJECTIVE: It remains unclear whether angiotensin II type 1 receptor antagonists (angiotensin receptor blockers [ARBs]) can inhibit neointimal hyperplasia after stent implantation in patients with coronary artery disease. The aim of this meta-analysis was therefore to evaluate the benefits of ARBs in patients after coronary stent implantation based on the currently available randomized controlled trials. METHODS: We conducted a pooled analysis of randomized controlled trials to compare outcomes after stent implantation in patients administered ARBs with those not administered ARBs. We searched Ovid/MEDLINE, EMBASE, and the ISI web of knowledge using the terms 'angiotensin receptor blocker,' 'renin angiotensin system inhibitor,' 'angiotensin receptor antagonist,' 'stent,' 'angiograph,' 'percutaneous coronary intervention (PCI),' and 'coronary artery disease.' Published meta-analyses, review articles, and editorials were reviewed for potential studies of interest. The inclusion criteria were randomized controlled trials published in English, with a follow-up period of 6 months, comparing the outcomes after coronary stent implantation with and without the administration of any kind of ARB, reporting at least one outcome of interest (restenosis rate and late lumen loss). Data abstraction included study design, patient characteristics, follow-up period, type of ARB, type of stent, restenosis rate, and late lumen loss. Fixed-effects models were used to calculate the pooled relative risk for the restenosis rate and the standardized mean difference for late lumen loss. RESULTS: Five studies were included, with a total number of 624 patients. Seventy-five of 314 patients in the ARB group were diagnosed with in-stent restenosis at the 6-month follow-up, compared with 87 of 310 patients in the control group (relative risk 0.85; 95% CI 0.65, 1.11; p = 0.23). Consistent with this, there was no significant difference in late lumen loss between the two groups (0.04 mm; 95% CI -0.15, 0.23; p = 0.66). CONCLUSION: There is no evident benefit with the use of an ARB in terms of inhibition of neointimal hyperplasia in patients after coronary stent implantation. PMID- 21619385 TI - Usefulness of ultrasonography for the diagnosis of difuse liver diseases. PMID- 21619386 TI - Prospective validation of two models for ultrasonographic diagnosis of cirrhosis. AB - OBJECTIVE: To perform a prospective validation and comparative analysis of two ultrasonographic diagnostic scores of cirrhosis in patients with silent liver disease. DESIGN: cross-sectional study, prospective and blind. ROC curves evaluated the diagnostic utility of: a) Bologna score (BS): assessment of liver surface nodularity and portal flow velocity, and b) Cadiz score (CS): assessment of liver echo structure, portal vein caliber and spleen area. Liver biopsy was considered the gold standard for the diagnosis of cirrhosis. PATIENTS: One hundred and thirteen patients, 76 men and 37 women, mean age 44 years old (range 18-73 years) referred for evaluation of chronic liver disease without clinical or biochemical evidence of advanced disease (absence of jaundice, ascites, encephalopathy, malnutrition or coagulopathy). RESULTS: Cirrhosis was diagnosed in 25 patients (22.1%). BS: sensitivity 84%, specificity 79.5%, area under the ROC curve 86.7%. CS: sensitivity 84%, specificity 89.8%, area under the ROC curve 92.4%. Portal vein was not displayed in 7 patients(6%) and portal flow velocity was not recorded in 13 (11.5%). These results agree with those obtained in the original articles developing both scores. There were no statistically significant differences between the two scores. Specificity reached 97% with joint use of both models, but sensitivity decreased to 72%. CONCLUSIONS: Presence or absence of cirrhosis in patients with silent liver disease can be established by Doppler ultrasound with high diagnostic accuracy. The joint use of both scores has high diagnostic specificity. Both diagnostic models are highly re-producible. PMID- 21619387 TI - Differences between pediatric and adult celiac disease. AB - INTRODUCTION: Celiac disease (CD) is a common autoimmune condition (involves 1-2% of the general population) that develops at any age in life but manifests differently in children and adults. OBJECTIVES: To analyze clinical differences in disease expression between both groups, as well as findings at the time of diagnosis. METHODS: A retrospective study of a series of patients diagnosed with CD during childhood (< 14 years) versus a series of adult patients (> 14 years). RESULTS: a total of 187 patients were included, of which 43 were children and 144 were adults. Among clinical manifestations in children classic presentation forms predominated -34 patients(79%) versus 20 adult patients (14%) (p < 0.001) (OR = 23.4;95% CI: 9.8-56.1). In contrast, atypical forms were predominant in the latter, and anemia was the most common finding in 61 patients (42%) versus 8 pediatric patients (19%) (p < 0.01). Adults had a greater diagnostic delay with a mean 10 +/- 9 years versus 1 +/- 2 years in children (p < 0.001). In adults, we found a higher frequency of associated autoimmune diseases (24.3 versus 9.3% in children) (p < 0.05). Regarding serum markers, TGt-2 was more commonly positive among children (88%) as compared to adults (31%) (p < 0.001); (OR = 21.4: 95% CI: 7.2-63.6). We found similar results with regard to the presence of villous atrophy, which was more common in children (95%) than in adults (33%) (p < 0.001) (OR = 41.0;95% CI: 9.5-76.7). As regards genetic markers, DQ2 was somewhat more common in children (97.7%) than in adults (90.3%) whereas DQ8 wasless common in children (2.3%) than in adults (9.7%), with no significant differences between groups. Patients negative for both markers were not included. CONCLUSIONS: Pediatric CD has clear differences when compared to adult CD, with classic forms predominating in the former, who also display a higher occurrence of positive serology and villous atrophy, and less diagnostic delay. In contrast, atypical forms predominate in the adult, with a lower occurrence of positive serology and milder histological forms. In these patients associated autoimmune conditions are more common and diagnostic delay is longer. PMID- 21619388 TI - Efficacy of intravenous iron in treating iron deficiency anaemia in patients with inflammatory bowel disease. Are there predictors of response? AB - INTRODUCTION: In inflammatory bowel disease (IBD) iron deficiency anaemia (IDA) is a very common disorder. Until recently,oral iron has been the mainstay therapy, nevertheless it has been associated with intolerance and noncompliance. Therefore, the goal of our study was to evaluate the efficacy of intravenous iron in IDA in IBD patients and the secondary aim was to investigate whether other potencial factors could influence in the response to the treatment. DESIGN: An open-label, prospective, consecutive, single centre study. MATERIAL AND METHODS: We performed our study in patients with ulcerative colitis (UC) or Crohn's disease (CD) with severe anaemia or intolerance with oral iron. All of them received intravenous sacarose iron and did biochemistry profile with hemoglobin (Hb). Moreover, the correlation with other variables was studied: age,sex, smoking habit, IBD type, previous surgery and type of surgery and other treatments. Response was defined as Hb increase of >= 2 g/dL or normalization of the levels. RESULTS: Fifty-four patients were included into the study, 34(63%) with UC y 20 (37%) with CD, 18 (33.3%) men and 36 wo-men (66.6%) and the average was 48 +/- 14 years. The total proportion of responders was 52% (SD +/- 05); 43% of the patients reached Hb >= 2 g/dl and y 9% of them normalized Hb. Only the utilization of 5-ASA was associated with low response to iron treatment (p < 0.05). CONCLUSIONS: Our study suggests that response to intravenous iron is achievable in the majority of patients with IBD and severe IDA or intolerance treatment with oral iron. Moreover, the patients with consumption of 5-ASA could had less response to the treatment. PMID- 21619389 TI - Surgical pathology associated with Meckel's diverticulum in a tertiary hospital: 12 year review. AB - OBJECTIVE: We want to present our experience about surgical pathology of Meckel's diverticulum by means of a retrospective study. MATERIAL AND METHODS: We report a group of patients of our Department of General and Abdominal Surgery and Paediatric Surgery of our Hospital with Meckel's diverticulum since January 1997 to January 2010. We report the clinical presentation, complementary test, interventions, and the postoperative follow up. RESULTS: 45 patients were operated in total, 33 of them in emergency surgery under the clinical form of acute abdominal pain; and the others 12 in programmed surgery, these cases came up more frequently like a clinical manifestations of latent abdominal pain, rectal bleeding and anemia. The complementary tests were so varied; abdominal ultrasounds were used in 63% of emergency cases and the 40% of programmed cases, in these patients,gammagraphy with Tc99 was the second test in frequency. Laparoscopy was used in 10 cases (22%). The main surgery technique used was diverticulectomy (82%). CONCLUSIONS: The presence of Meckel's diverticulum has to be clinically suspected in all patients with abdominal pain of unknow naetiology. Access to the abdominal cavity using routine laparoscopy provides essential information on the diagnosis and for the treatment. PMID- 21619390 TI - Spontaneous bacterial peritonitis. PMID- 21619391 TI - Tuboovarian abscess as unusual presentation of tubarian fistula secondary to sigmoid diverticulitis. PMID- 21619392 TI - Large asymptomatic type III paraesophageal hernia. PMID- 21619393 TI - Amyloidosis and Crohn's disease. AB - Secondary amyloidosis is a rare but serious complication of inflammatory bowel disease that may influence the prognosis even more than the underlying disease. Due to a better knowledge of the association of secondary amyloidosis to inflammatory bowel disease, early diagnosis of this complication is becoming more frequent, but its treatment continues to pose a challenge. We report 4 cases of patients with Crohn's disease and amyloidosis diagnosed in the inflammatory bowel disease Units of Toledo and Ciudad Real, which represent 0.68% of the patients with Crohn's disease of our health areas. There have been not cases of amyloidosis in patients with ulcerative colitis. In our 4 patients the secondary amyloidosis was clearly related to Crohn's disease,which was often of fistulising type. The predominant clinical picture of amyloidosis was nephrotic syndrome. The patients responded to medical and surgical treatment of Crohn's disease and colchicine, which improved renal function in all cases except in one who required kidney transplantation. PMID- 21619394 TI - [Acetaminophen and the liver]. PMID- 21619395 TI - Rare rectal mucocele mimic tumor following hemorrhoidectomy in an adult patient. PMID- 21619396 TI - Gallstone ileus with spontaneous resolution. PMID- 21619397 TI - Advanced esophageal squamous cell carcinoma in patient with lonstanding achalasia. PMID- 21619398 TI - [Systemic thromboembolic complications in a severe brout of ulcerative colitis]. PMID- 21619399 TI - Hepatitis C virus and hepatocellular carcinoma. PMID- 21619400 TI - Therapeutic management of emphysematous pancreatitis. PMID- 21619401 TI - Nerve injury during the transpsoas approach for lumbar fusion. AB - A lateral transpsoas approach to achieve interbody fusion in the lumbar spine using either the extreme lateral interbody fusion or direct lateral interbody fusion technique is an increasingly popular method to treat spinal disease. Dissection and dilation through the iliopsoas muscle places the lumbosacral plexus at risk for injury, but there is very limited information in the published literature about adverse clinical events resulting in postoperative motor deficits or reports of failure of electrophysiological monitoring to detect nerve injury. The authors present 2 cases of postoperative motor deficits following the transpsoas approach not detected by intraoperative monitoring, review the medical literature, and discuss strategies for complication avoidance. PMID- 21619402 TI - Cervical pneumatocyst. AB - The authors present the case of a 56-year-old right hand-dominant woman who was referred for chronic neck pain and a second opinion regarding a cervical lesion. The patient's pain was localized to the subaxial spine in the midline. She reported a subjective sense of intermittent left arm weakness manifesting as difficulty manipulating small objects with her hands and fingers. She also reported paresthesias and numbness in the left hand. Physical and neurological examinations demonstrated no abnormal findings except for a positive Tinel sign over the left median nerve at the wrist. Electromyography demonstrated bilateral carpal tunnel syndrome with no cervical radiculopathy. Cervical spine imaging demonstrated multilevel degenerative disc disease and a pneumatocyst of the C-5 vertebral body. The alignment of the cervical spine was normal. A review of the patient's cervical imaging studies obtained in 1995, 2007, 2008, and 2010 demonstrated that the pneumatocyst was not present in 1995 but was present in 2007. The lesion had not changed in appearance since 2007. At an outside institution, multilevel fusion of the cervical spine was recommended to treat the pneumatocyst prior to evaluation at the authors' institution. The authors, however, did not think that the pneumatocyst was the cause of the patient's neck pain, and cervical pneumatocysts typically have a benign course. As such, the authors recommended conservative management and repeated MR imaging in 6 months. Splinting was used to treat the patient's carpal tunnel syndrome. PMID- 21619403 TI - Interbody versus posterolateral fusion. PMID- 21619404 TI - Meta-analysis of instrumented posterior interbody fusion versus instrumented posterolateral fusion in the lumbar spine. AB - OBJECT: The authors compared the effectiveness of instrumented posterior lumbar interbody fusion (iPLIF) and instrumented posterolateral fusion (iPLF) for the treatment of low-back pain (LBP) due to degenerative lumbar disease. METHODS: Relevant randomized controlled trials (RCTs) and comparative observational studies through December 2009 were identified using a retrieval strategy of sensitive and specific searches. The study design, participant characteristics, interventions, follow-up rate and period, and outcomes were abstracted after the assessment of methodological quality of the trials. Analyses were performed following the method guidelines of the Cochrane Back Review Group. RESULTS: Nine studies were identified-3 RCTs and 6 comparative observational studies. No significant difference was found between the 2 fusion procedures in the global assessment of clinical outcome (OR 1.51, 95% CI 0.71-3.22, p = 0.29) and complication rate (OR 0.55, 95% CI 0.16-1.86, p = 0.34). Both techniques were effective in reducing pain and improving functional disability, as well as restoring intervertebral disc height. Instrumented PLIF was more effective in achieving solid fusion (OR 2.60, 95% CI 1.35-5.00, p = 0.004), a lower reoperation rate (OR 0.20, 95% CI 0.03-1.29, p = 0.09), and better restoration of segmental angle and lumbar lordotic angle than iPLF. There were no significant differences between the fusion methods regarding blood loss (weighted mean difference -179.63, 95% CI -516.42 to 157.15, p = 0.30), and operating time (weighted mean difference 8.03, 95% CI -45.46 to 61.53, p = 0.77). CONCLUSIONS: The authors' analysis provided moderate-quality evidence that iPLIF has the advantages of higher fusion rate and better restoration of spinal alignment over iPLF. No significant differences were identified between iPLIF and iPLF concerning clinical outcome, complication rate, operating time, and blood loss. PMID- 21619405 TI - Crescent sign on magnetic resonance angiography revealing incomplete stent apposition: correlation with diffusion-weighted changes in stent-mediated coil embolization of aneurysms. AB - OBJECT: Few data are available on how closely stents appose the luminal vessel wall in stent-mediated coil embolization of intracranial aneurysms and on the effect of incomplete stent apposition on procedural thromboembolic complications. METHODS: Postprocedural 3-T MR diffusion-weighted imaging and time-of-flight angiography were obtained in 58 patients undergoing stent-mediated coil embolization of aneurysms using the Enterprise closed-cell and Neuroform open cell self-expanding intracranial microstents. RESULTS: A distinctive semilunar signal pattern, identified using 3-T MR angiography, represented flow outside the confines of the stent struts in patients in whom Enterprise but not Neuroform devices were used. This pattern, designated as the crescent sign, was confirmed to correspond to incomplete stent apposition by use of high-resolution angiographic flat-panel CT scanning revealing flow ingress into and egress out of the isolated luminal wedge. The presence of the crescent sign was seen in 18 of 33 Enterprise-treated but in 0 of 25 Neuroform-treated cases, and was more likely in stents delivered in the tortuous internal carotid artery (p = 0.034). The crescent sign was strongly predictive of ipsilateral postprocedural lesions seen on diffusion-weighted imaging in the entire population (OR 18, 95% CI 4.33-74.8; p < 0.0001). In the Enterprise stent subset, ipsilateral lesions were detected on diffusion-weighted imaging in 15 (45%) of 33 cases; the crescent sign was seen in 12 (80%) of 15 patients with ipsilateral lesions on diffusion-weighted imaging, but in only 6 of 18 patients without lesions (OR 8, 95% CI 1.61-39.6; p = 0.006). CONCLUSIONS: Incomplete stent apposition is detectable on 3-T MR angiography as a crescent sign, and was found to be highly prevalent in Enterprise closed-cell design stents used to assist coil embolization of aneurysms. Incomplete stent apposition was also associated with periprocedural ipsilateral hyperintense lesions on diffusion-weighted imaging. These results identify an association between incomplete stent apposition and thromboembolic complications in stent mediated coil embolization of intracranial aneurysms. PMID- 21619406 TI - Cervical clearance. PMID- 21619407 TI - Angiographic features of "brain sag". AB - OBJECT: Cerebrospinal fluid hypotension, or "brain sag," is a recently described phenomenon most commonly seen following craniotomy for the clipping of ruptured aneurysms along with preoperative lumbar drain placement. The clinical features and CT findings have been previously described. Clinical presentation can be similar to and often mistaken for cerebral vasospasm. In this study, the authors report on the angiographic findings in patients with brain sag. METHODS: Five cases of brain sag were diagnosed (range 1-4 days) after the surgical treatment of ruptured aneurysms at the University of Illinois at Chicago. All patients met the clinical and CT criteria for brain sag. Admission cerebral angiograms and subsequent angiograms during symptoms of brain sag were obtained in all patients. In 3 patients, angiography was performed after the resolution of symptoms. RESULTS: In all 5 patients, the level of the basilar artery apex was displaced inferiorly with respect to the posterior clinoid processes during brain sag. This displacement was significant enough to create a noticeable kink in the basilar artery ("cobra sign") in 3 patients. Other angiographic findings included foreshortening or kinking of the intracranial vertebral artery. In all patients, the posterior cerebral arteries were displaced medially and inferiorly. Three patients were treated for simultaneous severe radiological vasospasm. In 4 patients, the brain sag was recognized, and the patients' conditions improved when they were placed flat or in the Trendelenburg position, at times combined with an epidural blood patch. Patients with follow-up angiography studies after the symptoms had resolved displayed a reversal of the angiographic features. CONCLUSIONS: Brain sag appears to be associated with characteristic angiographic features. Recognizing these features may help to diagnose brain sag as the cause of neurological deterioration in this patient population. PMID- 21619408 TI - Comparative effectiveness of using computed tomography alone to exclude cervical spine injuries in obtunded or intubated patients: meta-analysis of 14,327 patients with blunt trauma. AB - OBJECT: The current standard of practice for clearance of the cervical spine in obtunded patients suffering blunt trauma is to use CT and an adjuvant imaging modality (such as MR imaging). The objective of this study was to determine the comparative effectiveness of multislice helical CT alone to diagnose acute unstable cervical spine injury following blunt trauma. METHODS: The authors performed a meta-analysis of studies comparing modern CT with adjunctive imaging modalities and required that studies present acute traumatic findings as well as treatment for unstable injuries. Study quality, population characteristics, diagnostic protocols, and outcome data were extracted. Positive disease status included all injuries necessitating surgical or orthotic stabilization identified on imaging and/or clinical follow-up. RESULTS: Seventeen studies encompassing 14,327 patients met the inclusion criteria. Overall, the sensitivity and specificity for modern CT were both > 99.9% (95% CI 0.99-1.00 and 0.99-1.00, respectively). The negative likelihood ratio of an unstable cervical injury after a CT scan negative for acute injury was < 0.001 (95% CI 0.00-0.01), while the negative predictive value of a normal CT scan was 100% (95% CI 0.96-1.00). Global severity of injury, CT slice thickness, and study quality did not significantly affect accuracy estimates. CONCLUSIONS: Modern CT alone is sufficient to detect unstable cervical spine injuries in trauma patients. Adjuvant imaging is unnecessary when the CT scan is negative for acute injury. Results of this meta analysis strongly show that the cervical collar may be removed from obtunded or intubated trauma patients if a modern CT scan is negative for acute injury. PMID- 21619409 TI - Effect of concentration on the accuracy of convective imaging distribution of a gadolinium-based surrogate tracer. AB - OBJECT: Accurate real-time imaging of coinfused surrogate tracers can be used to determine the convective distribution of therapeutic agents. To assess the effect that a concentration of a Gd-based surrogate tracer has on the accuracy of determining the convective distribution, the authors infused different concentrations of Gd-diethylenetriamine pentaacetic acid (DTPA) in primates during MR imaging. METHODS: Five nonhuman primates underwent convective infusion (1 or 5 mM, 21-65 MUl) of Gd-DTPA alone, Gd-DTPA and (14)C-sucrose, or Gd-DTPA and (14)C-dextran into the bilateral striata. Animals underwent real-time MR imaging during infusion (5 animals) and autoradiographic analysis (2 animals). RESULTS: Gadolinium-DTPA could be seen filling the striata at either concentration (1 or 5 mM) on real-time MR imaging. While the volume of distribution (Vd) increased linearly with the volume of infusion (Vi) for both concentrations of tracer (1 mM: R(2) = 0.83; 5 mM: R(2) = 0.96), the Vd/Vi ratio was significantly (p < 0.0001) less for the 1-mM (2.3 +/- 1.0) as compared with the 5-mM (7.4 +/- 1.9) concentration. Autoradiographic and MR volumetric analysis revealed that the 5-mM concentration most accurately estimated the Vd for both small (sucrose [359 D], 12% difference between imaging and autoradiographic distribution) and large (dextran [70 kD], 0.2% difference) molecules compared with the 1-mM concentration (sucrose, 65% difference; dextran, 68% difference). CONCLUSIONS: The concentration of infused Gd-DTPA plays a critical role in accurately assessing the distribution of molecules delivered by CED. A 5-mM concentration of Gd-DTPA most accurately estimated the Vd over a wide range of molecular sizes. PMID- 21619410 TI - Preliminary evaluation of a novel intraparenchymal capacitive intracranial pressure monitor. AB - OBJECT: Intracranial pressure (ICP) monitors are currently based on fluid-filled, strain gauge, or fiberoptic technology. Capacitive sensors have minimal zero drift and energy requirements, allowing long-term implantation and telemetric interrogation; their application to neurosurgery has only occasionally been reported. The aim of this study was to undertake a preliminary in vitro and in vivo evaluation of a capacitive telemetric implantable ICP monitor. METHODS: Four devices were tested in air- and saline-filled pressure chambers; long-term capacitance-pressure curves were obtained. Devices implanted in a gel phantom and in a piglet were placed in a 3-T MR unit to evaluate MR compatibility. Four devices were implanted in a piglet neonatal hydrocephalus model; output was compared with ICP obtained through fluid-filled transduction and a strain-gauge ICP monitor. RESULTS: The capacitance-pressure relationship was constant over 4 weeks, suggesting minimal zero drift during this period. There were no temperature changes around the monitor. Signal loss at the sensor was minimal in both the phantom and the piglet. Over 114,000 measurements were obtained; the difference between mean capacitive ICP and fluid-transduced ICP was 1.8 +/- 1.42 mm Hg. The correlation between ICP from the capacitive sensor and fluid-filled transducer (r = 0.97, p < 0.0001) or strain-gauge monitor (r = 0.99, p < 0.0001) was excellent. In vivo monitoring was restricted to 48 hours due to problems with robustness in the clinical environment. CONCLUSIONS: This preliminary study demonstrates minimal long-term zero drift in vitro, good MR compatibility, and good correlation with other methods of ICP monitoring in vivo in the short term. Further long-term in vivo study is required. PMID- 21619411 TI - Neurosurgical intervention in the diagnosis and treatment of Balamuthia mandrillaris encephalitis. AB - The authors describe the unique presentation of Balamuthia mandrillaris encephalitis in a kidney donor and two recipients. All three patients suffered acute clinical deterioration, with radiological or clinical evidence of intracranial hypertension. Brain autopsy in the donor and an urgent brain biopsy in a recipient established the diagnosis. First-tier therapy, with mannitol and CSF drainage, successfully treated the intracranial hypertension in both recipients, while administration of a combination of antiamebic drugs was associated with survival in the second recipient. For both recipients, neurosurgical management played a critical role in the rapid diagnosis and treatment of Balamuthia mandrillaris encephalitis. PMID- 21619412 TI - Brain sag. PMID- 21619413 TI - Convection-enhanced delivery. PMID- 21619414 TI - Involvement of BMPR2 in the protective effect of fluoxetine against monocrotaline induced endothelial apoptosis in rats. AB - Mutations in bone morphogenetic protein (BMP) receptor II (BMPR2) are associated with the apoptosis of the pulmonary artery endothelial cells and the loss of the pulmonary small vessels. The present study was designed to investigate the involvement of BMPR2 in the protective effect of fluoxetine against monocrotaline (MCT)-induced endothelial apoptosis in rats. Models of pulmonary arterial hypertension in rats were established by a single intraperitoneal injection of MCT (60 mg/kg). Fluoxetine (2 and 10 mg/kg) was intragastrically administered once a day. After 21 days, MCT caused pulmonary hypertension, right ventricular hypertrophy, and pulmonary vascular remodeling and significantly reduced the BMPR2 expression in lungs and pulmonary arteries. Fluoxetine dose-dependently inhibited MCT-induced pulmonary arterial hypertension and effectively protected the lungs against MCT-induced endothelial apoptosis, reduction in the number of alveolar sacs, and loss of the pulmonary small vessels. Fluoxetine reversed the expression of cyclic guanosine 3',5'-monophosphate-dependent kinase I, BMPR2, phospho-Smad1, beta-catenin, and reduced the expression of caspase 3 in rat lungs. These findings suggest that BMPR2 is probably involved in the protective effect of fluoxetine against MCT-induced endothelial apoptosis in rats. PMID- 21619415 TI - Arachidonic acid epoxygenase and 12(S)-lipoxygenase: evidence of their concerted involvement in ductus arteriosus constriction to oxygen. AB - Oxygen promotes closure of the ductus arteriosus at birth. We have previously presented a scheme for oxygen action with a cytochrome P450 (CYP450) hemoprotein and endothelin-1 (ET-1) being, respectively, sensor and effector, and a hypothetical monooxygenase product serving as a coupling link. We have also found in the vessel arachidonic acid (AA) 12(S)-lipoxygenase (12-lipoxygenase) undergoing upregulation at birth. Here, we examined the feasibility of a sensor to-effector messenger originating from AA monooxygenase and 12-lipoxygenase pathways. The epoxygenase inhibitor, N-methylsulfonyl-6-(2-)hexanamide, suppressed the tonic contraction of ductus to oxygen. A similar effect was obtained with 12-lipoxygenase inhibitors baicalein and PD 146176. By contrast, none of the inhibitors modified the endothelin-1 contraction. Furthermore, an AA omega-hydroxylation product, 20-hydroxyeicosatetraenoic acid (20-HETE), reportedly responsible for oxygen contraction in the systemic microvasculature, had no such effect on the ductus. We conclude that AA epoxygenase and 12 lipoxygenase jointly produce a hitherto uncharacterized compound acting as oxygen messenger in the ductus. PMID- 21619416 TI - Protective effect of L-citrulline against acute gastric mucosal lesions induced by ischemia-reperfusion in rats. AB - The present study investigated the protective effect of L-citrulline on gastric mucosal injury induced by ischemia-reperfusion (IR) in rats. Under anesthesia, the celiac artery was clamped for 30 min, and then the clamp was removed for 60 min reperfusion. Sixty minutes before ischemia, L-citrulline was administered intragastrically at doses of 300, 600, and 900 mg/kg. After the experiment, the stomachs were removed for biochemical and histological examinations. Pretreatment with L-citrulline (300, 600, and 900 mg/kg) significantly ameliorated the gastric damage caused by IR. Moreover, L-citrulline prevented the production of lipid peroxidation and inhibited the increase of myeloperoxidase activity. The elevation in total nitric oxide synthase (NOS) activity, inducible NOS activity, and inducible NOS protein expression as well as the decrease in constitutive NOS activity and gastric mucus level in the gastric mucosa induced by IR were significantly prevented. However, the protective effect mediated by L-citrulline was significantly antagonized by coadministration of L-nitroarginine methyl ester (10 mg/kg, s.c.). These results suggest that part of the mechanism of gastric protection by L-citrulline might be through inhibiting neutrophil infiltration and preserving gastric mucus synthesis and secretion in rats, functions that are closely related to the maintenance of constitutive NOS activity. PMID- 21619417 TI - Chromophobe renal cell carcinoma in Iceland: an epidemiological and clinicopathological study. AB - OBJECTIVE: Numerous studies have suggested that the rare chromophobe renal cell carcinoma (CRCC) has a more favourable prognosis than the other more common subtypes of RCC, clear cell RCC (CCRCC) and papillary RCC (PRCC). These studies have, however, usually involved selected patient cohorts and not whole populations. This study compared CRCC patients with patients with the other two major histological subtypes and established a population-based age-standardized incidence rate (ASR). MATERIAL AND METHODS: Of 828 histopathologically confirmed RCCs diagnosed between 1971 and 2005 in Iceland, 15 CRCC cases were identified. Histological material was reviewed, the TNM system was used for staging and cancer-specific survival was estimated. Univariate and multivariate analysis was used to compare CRCC to both CCRCC (n = 740) and PRCC (n = 66). Mean follow-up was 6.7 years. RESULTS: CRCC accounted for 1.8% of RCCs, the ASR being 0.17/100,000 per year. Compared to other subtypes, CRCC was detected incidentally less often (7% vs 29%, p = 0.02), but was more often diagnosed at lower stages (73% vs 45% at stage I + II, p < 0.001). One patient had synchronous metastasis and another developed recurrent CRCC; both died of CRCC. Five-year survival for CRCC, CCRCC and PRCC was 86%, 59% and 50%, respectively (p = 0.004). After correcting for TNM stage (odds ratio 1.98), multivariate analysis did not indicate that CRCC subtype was an independent predictive factor for survival. CONCLUSION: CRCC is a rare neoplasm with an ASR of 0.17/100,000 per year. These tumours often present with symptoms despite being at lower stages than the other RCC subtypes. The more favourable survival of the CRCC subtype appears to be explained by these tumours being diagnosed at low stages. These findings may suggest that CRCC has a different biological behaviour. PMID- 21619418 TI - Perceptions of pediatric physical therapists and physical educators on classifying learning styles of children and adolescents with cerebral palsy. AB - The purpose of this study was to examine professionals' perceptions on classifying learning styles in the context of teaching motor activities to children and adolescents with cerebral palsy (CP). The participants were 21 pediatric physical therapists (PPTs) and seven physical educators (PEs) in three schools for special education in The Netherlands. All participants were introduced to the key descriptions of two existing learning style instruments (Kolb's Learning Style Inventory and Myers-Briggs Type Indicator), applied them to children and adolescents with CP, and reported their perceptions in written surveys. This study had a mixed-methods design. Quantitative and qualitative data analyses showed that PPTs and PEs are mostly positive about the idea of classifying learning styles in the context of teaching motor activities to children and adolescents with CP, giving three main reasons: individual approach, professional communication, and treatment awareness. Additionally, qualitative data analysis showed that the key descriptions of the two learning style instruments were not feasible as classifications for children and adolescents with CP. It is therefore recommended that other learning style classification instruments should be explored and that possibly a new learning style classification instrument should be developed in the context of teaching motor activities to children and adolescents with CP. PMID- 21619419 TI - The liquid overlay technique is the key to formation of co-culture spheroids consisting of primary osteoblasts, fibroblasts and endothelial cells. AB - BACKGROUND AIMS: The 3-dimensional (3-D) culture of various cell types reflects the in vivo situation more precisely than 2-dimensional (2-D) cell culture techniques. Spheroids as 3-D cell constructs have been used in tumor research for a long time. They have also been used to study angiogenic mechanisms, which are essential for the success of many tissue-engineering approaches. Several methods of forming spheroids are known, but there is a lack of systematic studies evaluating the performance of these techniques. METHODS: We evaluated the performance of the hanging drop technique, carboxymethyl cellulose technique and liquid overlay technique to form both mono- and co-culture spheroids consisting of primary osteoblasts, fibroblasts and endothelial cells. The performance of the three techniques was evaluated in terms of rate of yield and reproducibility. The size of the generated spheroids was determined systematically. RESULTS: The liquid overlay technique was the most suitable for generating spheroids reproducibly. The rate of yield for this technique was between 60% and 100% for monoculture spheroids and 100% for co-culture spheroids. The size of the spheroids could be adjusted easily and precisely by varying the number of seeded cells organized in one spheroid. The formation of co-culture spheroids consisting of three different cell types was possible. CONCLUSIONS: Our results show that the most suitable technique for forming spheroids can vary from the chosen cell type, especially if primary cells are used. Co-culture spheroids consisting of three different cell types will be used to study angiogenic phenomena in further studies. PMID- 21619420 TI - Wireless biopotential acquisition system for portable healthcare monitoring. AB - A complete biopotential acquisition system with an analogue front-end (AFE) chip is proposed for portable healthcare monitoring. A graphical user interface (GUI) is also implemented to display the extracted biopotential signals in real-time on a computer for patients or in a hospital via the internet for doctors. The AFE circuit defines the quality of the acquired biosignals. Thus, an AFE chip with low power consumption and a high common-mode rejection ratio (CMRR) was implemented in the TSMC 0.18-MUm CMOS process. The measurement results show that the proposed AFE, with a core area of 0.1 mm(2), has a CMRR of 90 dB, and power consumption of 21.6 MUW. Biopotential signals of electroencephalogram (EEG), electrocardiogram (ECG) and electromyogram (EMG) were measured to verify the proposed system. The board size of the proposed system is 6 cm * 2.5 cm and the weight is 30 g. The total power consumption of the proposed system is 66 mW. PMID- 21619421 TI - Assessment criteria for evaluating the stability and position of the centre of gravity on a balance training platform: a simulation with Matlab(r). AB - The anterior-posterior and medial-lateral tipping values of a training platform within the scope of a balance training activity should be evaluated in regard to position and the changing position of the centre of gravity (COG). Data streams are simulated and modulated with the help of the signal processing programme Matlab(r). These data streams are evaluated using existing balance formulas and a specially developed formula. Active Balance Index referring to the zero point (ABI(0)) allows an assertion about the magnitude of the mean tipping angle with the magnitude origin in the centre of the training platform. Active Balance Index referring to the arithmetical mean COG (ABI(mean)) enables an assertion about the mean tipping angle with the magnitude origin in the COG mean position. The deviation of the mean COG from absolute balance which is projected onto the platform is calculated by the Active Balance Ratio (ABR). The Active Balance Index (ABI) in combination with the ABR seems to be an adequate parameter for the evaluation of COG stability on a training platform. PMID- 21619423 TI - CD30: an important new target in hematologic malignancies. AB - CD30 is abundantly and selectively expressed on the surface of Hodgkin Reed Sternberg cells, anaplastic large cell lymphomas (ALCLs), and other lymphoid malignancies as well as on several non-lymphoid malignancies including selected germ cell tumors. Expression of CD30 on normal cells is highly restricted, thereby allowing differential targeting of malignant cells. CD30, a member of the tumor necrosis factor (TNF)-receptor family has pleiotropic biologic functions, and antibodies targeting CD30 and other TNF family receptors can exhibit both agonistic and antagonistic signaling functions. Recently, antibody-drug conjugates targeting CD30, such as brentuximab vedotin, have shown striking activity in phase I and II trials, with manageable toxicity. This has defined an important emerging role for targeting of CD30 in the setting of Hodgkin lymphoma, ALCL, and possibly other CD30+ malignancies. PMID- 21619424 TI - Quantiferon test for tuberculosis screening in sarcoidosis patients. AB - BACKGROUND: Tumour necrosis factor-alpha (TNF-alpha) inhibitors have been introduced in the treatment of refractory sarcoidosis. These biologics may reactivate latent tuberculosis infection (LTBI). Despite its known limitations, the tuberculin skin test (TST) is currently used for the diagnosis of LTBI in Danish sarcoidosis patients. We report the results of a screening using the interferon-gamma release assay (IGRA) QuantiFERON TB Gold (QFN) for the diagnosis of LTBI. We aimed to assess whether the QFN is reliable for diagnosing LTBI among sarcoidosis patients and if results are influenced by disease activity or immunosuppressive treatment. METHODS: A prospective study was performed from 2005 to 2007 among sarcoidosis patients who were candidates for TNF-alpha inhibitor treatment. Information on immunosuppressive treatment was obtained from the medical records. Disease activity was assessed by biochemistry, chest roentgenograms and pulmonary function tests. The predictive value of QFN results was evaluated by follow-up in the Danish National Tuberculosis Registry. RESULTS: A total of 44 sarcoidosis patients (22 men) with a median age of 39 y (range 25 59 y) were enrolled; 93% had a negative QFN test result and 7% had an indeterminate result. Forty-three percent had disease activity and 57% (n = 25) received immunosuppressive treatment. There was no significant difference in QFN interferon-gamma response between subjects with or without disease activity (p > 0.4) and between treated vs non-treated patients (p > 0.5). At follow-up using the Danish tuberculosis registry, there was no occurrence of tuberculosis among study participants. CONCLUSIONS: The predictive value of the QFN seems good among Danish sarcoidosis patients and the results appear to be unaffected by sarcoidosis disease activity and immunosuppressive treatment. PMID- 21619425 TI - Pharmacogenomics and personalized medicine in clinical practice. AB - The Santorini Conference on prospective biology, genomics and pharmacogenomics occurs every 2 years. On 30 September to 2nd October 2010, the fifth meeting in this series took place in Santorini, Greece. This conference has established a tradition of organizing a workshop each time to address the most recent developments and key issues in pharmacogenomics. This year, the workshop was chaired by Bryan Dechairo and Alain Huriez, and was titled 'Pharmacogenomics and personalized medicine in clinical practice'. PMID- 21619426 TI - Pharmacogenetics of drug transporters in the enterohepatic circulation. AB - This article summarizes the impact of the pharmacogenetics of drug transporters expressed in the enterohepatic circulation on the pharmacokinetics and pharmacodynamics of drugs. The role of pharmacogenetics in the function of drug transporter proteins in vitro is now well established and evidence is rapidly accumulating from in vivo pharmacokinetic studies, which suggests that genetic variants of drug transporter proteins can translate into clinically relevant phenotypes. However, a large amount of conflicting information on the clinical relevance of drug transporter proteins has so far precluded the emergence of a clear picture regarding the role of drug transporter pharmacogenetics in medical practice. This is very well exemplified by the case of P-glycoprotein (MDR1, ABCB1). The challenge is now to develop pharmacogenetic models with sufficient predictive power to allow for translation into drug therapy. This will require a combination of pharmacogenetics of drug transporters, drug metabolism and pharmacodynamics of the respective drugs. PMID- 21619427 TI - SNPs in genes encoding G proteins in pharmacogenetics. AB - Heterotrimeric guanine-binding proteins (G proteins) transmit signals from the cell surface to intracellular signal cascades and are involved in various physiological and pathophysiological processes. Polymorphisms in the genes GNB3 (encoding the Gbeta3 subunit), GNAS (encoding the Galphas subunit) and GNAQ (encoding the Galphaq subunit) have been the primary focus of investigation. Polymorphisms in these genes could be associated with different complex phenotypes underlining that alterations in G-protein signaling can cause multiple disorders. G proteins present a point of convergence or 'bottleneck' between various receptors and effectors, thus making them a sensible tool for pharmacogenetic studies. The pharmacogenetic studies performed to date mostly demonstrate an association between G-protein polymorphisms and response to therapy or occurrence of adverse drug effects. Therefore, polymorphisms in genes encoding G-protein subunits may help to individualize drug treatment in various diseases with regard to both efficacy and safety. PMID- 21619428 TI - Transcriptional regulation and pharmacogenomics. AB - Interindividual variable drug response is correlated with sequence alterations in genes encoding drug-metabolizing enzymes and transporters, affecting drug absorption, distribution, metabolism and excretion. This variable drug response may have an impact on disease therapeutic outcomes, tolerance to adverse drug reactions and even survival. Sequence alterations may occur not only within the coding region of a gene, but in its regulatory elements too, affecting gene transcription and gene-expression levels. Here, we provide a compilation of the current knowledge of pharmacogenomics related to transcription, with a focus on the effect of SNPs and short tandem repeats residing in cis-regulatory elements of 11 genes encoding for drug-metabolizing enzymes and drug transporters. In addition, we comment on two genes encoding enzymes that are drug targets themselves. Finally, we briefly discuss the currently available methodologies for clinically assessing pharmacogenomic profiles, which could potentially in the future facilitate drug treatment-individualization via the identification of molecular signatures in specific patient groups. PMID- 21619429 TI - Pharmacogenomic biomarkers: validation needed for both the molecular genetic mechanism and clinical effect. AB - Drug therapy can be ineffective or cause adverse reactions in a subset of patients. Pharmacogenomic biomarkers afford the opportunity to optimize an individual's therapy. Yet, few tests are currently part of standard care. To validate biomarkers, clinical replication studies are essential. Equally important, but less appreciated, the genetic mechanisms must also be understood to facilitate translation into clinical use. Representing main contributors to genetic variability, regulatory polymorphisms in particular are still poorly studied (e.g., 5-HTTLPR). This article focuses on molecular and functional diversity of genetic biomarkers, as a guide to optimal use in personalized medicine. PMID- 21619430 TI - Availability of pharmacogenetic and pharmacogenomic information in anticancer drug monographs in France: personalized cancer therapy. AB - AIM: To determine the availability of pharmacogenetic and pharmacogenomic information for healthcare professionals in France during 2009 for anticancer drugs. MATERIALS & METHODS: We searched in the informatic version of the VIDAL dictionary which is currently used by healthcare professionals in France. We then compared this with data available in the PubMed database. RESULTS: Among the 109 anticancer molecules available in France during 2009, 13 have pharmacogenomic or pharmacogenetic information in their monographs. In the scientific literature, we found numerous pharmacogenomic and pharmacogenetic biomarkers concerning 43 of the 109 anticancer agents. Some are pharmacogenomic biomarkers related to drug effectiveness, others are pharmacogenetic biomarkers related to drug toxicity. CONCLUSION: We believe that the lack of pharmacogenomic and pharmacogenetic information in drug monographs reflects the relative newness of the discipline. However, pharmacogenetics and pharmacogenomics can offer valuable information for improving the safety of drugs, reducing toxicity and predicting nonresponders. The drugs might then be incorporated into clinical practice through several strategies, including increased drug labeling and better education of healthcare professionals. PMID- 21619431 TI - Networks medicine: from reductionism to evidence of complex dynamic biomolecular interactions. PMID- 21619432 TI - Drugs, diabetes and pharmacogenomics: the road to personalized therapy. PMID- 21619437 TI - Telmisartan provides protection against development of impaired vasodilation independently of metabolic effects in SHRSP.Z-Lepr(fa)/IzmDmcr rats with metabolic syndrome. AB - Metabolic syndrome is known to facilitate the development of cardiovascular disease. We have demonstrated that mesenteric arteries of SHRSP.Z Lepr(fa)/IzmDmcr (SHRSP-fatty) rats with metabolic syndrome display an impaired vasorelaxation response mediated by nitric oxide. We examined whether the condition could be alleviated by treatment with telmisartan, an angiotensin II type 1 (AT1) receptor antagonist with PPAR-gamma-activating properties and compared the results with those from pioglitazone, a PPAR-gamma agonist. Telmisartan (5 mg.kg(-1).day(-1)) or pioglitazone (2.5 mg.kg(-1).day(-1)) was orally administered to male SHRSP-fatty rats for 8 weeks. Serum triglyceride and cholesterol levels were determined, and the oral glucose tolerance test was performed to evaluate insulin resistance. Vasodilations in response to acetylcholine and nitroprusside were determined by wire myographs under isometric tension conditions, protein expressions of soluble guanylyl cyclase in mesenteric arteries by Western blotting, and the contents of 3-nitrotyrosine in aortas by high-performance liquid chromatography with electrochemical detection. Telmisartan exerted antihypertensive effects, while pioglitazone ameliorated metabolic abnormalities in SHRSP-fatty rats. Telmisartan increased acetylcholine- and nitroprusside-induced relaxation and soluble guanylyl cyclase protein expression in mesenteric arteries and reduced 3-nitrotyrosine content in aortas. Pioglitazone displayed no such alleviating effects on vascular functions. These findings indicate that telmisartan protects against vasodilation disturbance through anti-oxidative and -nitrative stress independently of metabolic effects in SHRSP-fatty rats with metabolic syndrome. PMID- 21619438 TI - A clinical study of the subjective visual vertical during unilateral centrifugation and static tilt. AB - CONCLUSION: The present study demonstrates that various response patterns of subjective visual vertical (SVV) can be identified during unilateral centrifugation (UC). It is proposed that these response types correspond to different degrees of compensation after disease. This is advantageous for monitoring the effect of rehabilitative measures and is useful in medico-legal issues. It also emerges that diagnosis of unilateral utricle function requires the determination not only of asymmetry ratio but also offset of SVV estimates. OBJECTIVES: A retrospective clinical study of SVV test results was performed to establish a classification and model of response types in patients with suspected otolith disorder. METHODS: SVV measurements were made in 473 patients recruited from the dizziness clinic. A control group of healthy subjects (n = 43) was tested with the same protocol. Testing with bilateral stimulation (stationary upright, 15 degrees , 30 degrees tilt) and UC was performed. A mathematical model for the UC results was developed. RESULTS: During UC testing 61% of the patients showed an asymmetric response indicating a unilateral utricular hypofunction/dysfunction. These results could be classified into three subgroups, indicating different degrees of compensation. The model parameters can be adapted to reflect this classification. PMID- 21619439 TI - The insertion trajectory in cochlear implantation - comparison between two approaches. AB - CONCLUSION: The suprameatal approach (SMA) for cochlear implantation is a safe procedure and is at least comparable to the classic mastoidectomy-posterior tympanotomy approach (MPTA) regarding the possibility of reducing electrode insertion trauma. OBJECTIVES: To compare the trajectory in the SMA with insertion through cochleostomy, to the MPTA with round window insertion. METHODS: Nine temporal bones were implanted by both techniques, and the point of first contact of a precurved electrode was compared. RESULTS: With the SMA, in all bones, the point of first contact was the inferior wall of the scala tympani and insertion was into the scala tympani. In the MPTA, in five of the bones, the point of first contact was the modiolus, the osseous spiral lamina or the basilar membrane. PMID- 21619440 TI - Performance of the new Platelia Candida Plus assays for the diagnosis of invasive Candida infection in patients undergoing myeloablative therapy. AB - The performance of the new Platelia Candida Antigen Plus (Ag Plus) and Antibody Plus (Ab Plus) assays (Biorad Laboratories, France) was evaluated using a collection of serum samples obtained from 21 patients with microbiologically proven invasive candidiasis and 30 control patients who were being treated with myeloablative chemotherapy, and the data compared with that obtained with the earlier version of the Platelia Candida assays (Ag and Ab), and 1,3-beta-D-glucan (BG) detection systems. The sensitivity of the Ag Plus and Ab Plus assays in the per patient analysis ranged from 55-70% and from 30-64% for patients with less than 15 days of neutropenia and more than 15 days of neutropenia, respectively. Sensitivity and time to detection of these new assays was not significantly better than of the conventional Platelia Candida tests. However, the specificity of the Ag-Plus assay was reduced by approximately 50% as compared to the Ag assay. Logistic regression analysis showed that this was probably due to the fact that circulating mannan was also being detected by the Ag Plus assay in patients with superficial candidiasis. Further studies are needed to confirm our results and to determine the place of the Platelia Ag Plus and Ab Plus assays in the management of haematology patients at risk for Candida infections. PMID- 21619441 TI - Effects of policies to restrict malt liquor sales on neighborhood crime. AB - We examined the effectiveness of malt liquor sales restrictions adopted in 2005 in three liquor stores in a large Midwestern U.S. city. We hypothesized that the restrictions would be associated with crime reductions in adjacent neighborhoods. Using Poisson regression modeling, we compared crime rates two years prior to, and two years following policy adoption. Findings were mixed; malt liquor restrictions were associated with reductions in disorderly conduct citations, but increases in larceny/theft, beyond citywide trends. Limitations and implications of our study are discussed, and further research suggested. The study was funded by the U.S. Centers for Disease Control and Prevention. PMID- 21619442 TI - Determination of sleep quality and fatigue level of patients receiving continuous ambulatory peritoneal dialysis in Turkey. AB - OBJECTIVE: This descriptive study was performed to determine sleep quality, fatigue levels and related factors in patients receiving continuous ambulatory peritoneal dialysis (CAPD) treatment, as well as the effect of their sleep quality on fatigue. MATERIAL AND METHODS: Study data were accrued at the CAPD unit of a University Hospital nephrology service between 1 Marchand 30 June 30. Questionnaire forms, Pittsburgh Sleep Quality Indexand Piper Fatigue Scalewere used for data collection. Mann-Whitney U test, Kruskal-Wallis variance analysis, ttest, one-way analysis of variance and Pearson correlation coefficient for independent groups were used for the evaluation of data. RESULTS: Mean total fatigue and sleep quality scores (+/-1 SD) were 6.38 +/- 2.18 and 8.17 +/- 3.02, respectively. Patients in the study had severe fatigue and low sleep quality. CONCLUSIONS: Fatigue, sleep quality and related factors were determined in patients receiving peritoneal dialysis. The results of this study will be helpful in guiding future nursing methods for CAPD patients who experience fatigue and sleep problems. PMID- 21619443 TI - Changing drinking styles in Denmark and Finland. Fragmentation of male and female drinking among young adults. AB - A traditional heavy intoxication-oriented drinking style, "heroic drinking," is a central drinking practice in Denmark and Finland, especially among men. However, it seems that another drinking style leading to intoxication, "playful drinking," has become more prevalent in Denmark as well as in Finland. Playful drinking is characterized by self-presentations in diverse forms of game situations in which you need to play with different aspects of social and bodily styles. We approach the positions of heroic drinking and playful drinking among young adults (between 17 and 23 years) in Denmark and Finland by analyzing how they discuss these two drinking styles in focus groups (N = 16). PMID- 21619444 TI - Heavy drinking, perceived discrimination, and immigration status among Filipino Americans. AB - Filipino American drinkers (N = 1,443) in Honolulu and San Francisco were selected from the 1998-1999 Filipino American Community Epidemiological Survey to examine the association between perceived discrimination and heavy drinking behavior by immigration status. Results indicate that living in San Francisco, lower religious participation, and higher perceived discrimination were associated with increased odds for heavy drinking among US-born individuals, whereas being male was a risk factor among foreign-born individuals. Thus, perceived discrimination and immigration status should be considered when designing prevention and intervention strategies to address heavy drinking behavior in this population. PMID- 21619445 TI - Effects of a community prevention intervention on public awareness, knowledge, and risk perception of club drug use by youth in Taiwan. AB - This study evaluated the effects of mobilizing community coalitions and implementing prevention intervention concerning public awareness, knowledge, and perception of the risks associated with club drug use by Taiwanese urban youth. A quasi-experimental design was used. Three communities in Taipei city were included in the present study. A total of 328 residents successfully participated in the baseline survey (April 2008), and 276 residents were successfully interviewed for the follow-up survey (September 2008). The generalized estimating equation (GEE) method was used. The percentage of the intervention respondents who reported having seen or heard antidrug messages increased significantly between the time of the baseline survey (63.7%) and the follow-up survey (77.4%), while the percentage of attendance at antidrug events increased from 23.1% to 38.7% during the same time interval. In addition, community knowledge and perception of the problem of club drug use by youth rose significantly between the baseline and the follow-up in the intervention communities. The study's limitations are noted. PMID- 21619446 TI - Underestimations of blood alcohol concentration predict event-specific negative consequences. AB - The current study sought to establish in vivo misperception of blood alcohol concentration (BAC) as a predictor of event-specific alcohol-consumption-related negative consequences. During spring 2010, 225 (56.4% male) college students, who had consumed at least one alcoholic drink within the 2 hr prior to assessment, completed a questionnaire, gave a breath sample to assess breath alcohol content, and later completed a follow-up questionnaire. Underestimation of BAC was predictive of event-specific, alcohol-consumption-related negative consequences, over and above other factors including total drinks consumed. This study highlights the need for more focused BAC education strategies at American universities. PMID- 21619447 TI - Novel electrostatic mechanism for mode of action by N-acetylated proteins: cell signaling and phosphorylation. AB - Although extensive literature exists for N-acetylated proteins, scant knowledge is available concerning resultant mode of action. This review presents a novel mechanism based on electrostatics and cell signaling. There is substantial increase in the amide dipole and electrostatic field (EF) in contrast with the primary amino of the lysine precursor. The EF might serve as a bridge in electron transfer and cell signaling or energetics may play a role. The relationship between N-acetylation and phosphorylation is addressed. EFs may be important in the case of phosphates. Involvement of cell signaling is addressed including mechanistic aspects. As is the case for many aspects of bioaction, an integrated approach involving electrochemistry and cell signaling seems reasonable. PMID- 21619448 TI - GABRB3 gene expression increases upon ethanol exposure in human embryonic stem cells. AB - Ionotropic receptors are the target for most mood-defining compounds. Chronic exposure to ethanol (EtOH) alters receptor-mediated responses and the numbers of these channels and specific subunits; as well as induces anxiolytic, sedative, and anesthetic activity in the human brain. However, very little is known regarding the effects of EtOH on ionotropic receptor transcription during early human development (preimplantation). Using two separate human embryonic stem cell lines the study shows that low amounts of EtOH (20 mM) alters transcription of the ionotropic subunit GABRB3. Changes in ionotrophic receptor expression influence the central nervous system development and have been shown to produce brain abnormalities in animal models. These results suggest that low concentrations of EtOH can alter ionotropic receptor transcription during early human development (preimplantation), which may be a contributing factor to the neurological phenotypes seen in fetal alcohol spectrum disorder (FASD). PMID- 21619449 TI - Investigation of carvedilol-evoked Ca2+ movement and death in human oral cancer cells. AB - The effect of carvedilol on cytosolic free Ca2+ concentrations ([Ca2+](i)) in OC2 human oral cancer cells is unknown. This study examined if carvedilol altered basal [Ca2+](i) levels in suspended OC2 cells by using fura-2 as a Ca2+-sensitive fluorescent probe. Carvedilol at concentrations between 10 and 40 uM increased [Ca2+](i) in a concentration-dependent fashion. The Ca2+ signal was decreased by 50% by removing extracellular Ca2+. Carvedilol-induced Ca2+ entry was not affected by the store-operated Ca2+ channel blockers nifedipine, econazole, and SK&F96365, but was enhanced by activation or inhibition of protein kinase C. In Ca2+-free medium, incubation with the endoplasmic reticulum Ca2+ pump inhibitor thapsigargin did not change carvedilol-induced [Ca2+](i) rise; conversely, incubation with carvedilol did not reduce thapsigargin-induced Ca2+ release. Pretreatment with the mitochondrial uncoupler carbonylcyanide m chlorophenylhydrazone (CCCP) inhibited carvedilol-induced [Ca2+](i) release. Inhibition of phospholipase C with U73122 did not alter carvedilol-induced [Ca2+](i) rise. Carvedilol at 5-50 uM induced cell death in a concentration dependent manner. The death was not reversed when cytosolic Ca2+ was chelated with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid acetoxymethyl ester (BAPTA/AM). Annexin V/propidium iodide staining assay suggests that apoptosis played a role in the death. Collectively, in OC2 cells, carvedilol induced [Ca2+](i) rise by causing phospholipase C-independent Ca2+ release from mitochondria and non-endoplasmic reticulum stores, and Ca2+ influx via protein kinase C-regulated channels. Carvedilol (up to 50 MUM) induced cell death in a Ca2+-independent manner that involved apoptosis. PMID- 21619450 TI - Structure-function relationships of the human bitter taste receptor hTAS2R1: insights from molecular modeling studies. AB - Bitter taste receptors (T2Rs) belong to G-protein-coupled receptors (GPCRs). Despite extensive studies, the precise mechanisms of GPCR activation are still poorly understood. In this study, the models of the human bitter taste receptor hTAS2R1 alone and in complex with various ligands were constructed on the basis of template-based modeling and molecular docking. Then these models were subjected to all-atom molecular dynamics (MD) simulations in explicit lipid bilayers. The binding pocket of hTAS2R1 is mainly formed by transmembrane helix (TM) III, TM V, TM VI, and TM VII. Most of the residues contributing to ligand binding are positionally conserved comparing with other hTAS2Rs. By comparing the final conformations obtained by extensive MD simulations, we identified the changes in the transmembrane helices and the intra- and extracellular loops, which were expected to initiate the activation of the receptor. The intracellular loop II (ICL2) and TM III were found to play prominent roles in the process of activation. We proposed that a set of interactions between the aromatic Phe115 in the middle of ICL2 and three residues (Tyr103, Lys106, and Val107) at the cytoplasmic end of TM III may serve as a conformational switch of hTAS2R1 activation. All of the residues involved in the switch are highly conserved among T2Rs. This indicates that the control switch we proposed may be universal in T2Rs. Besides, our results also suggest that the formation of a short helical segment in ICL2 may be necessary for the activation of hTAS2R1. PMID- 21619451 TI - Flow cytometric analysis of STAT5 phosphorylation and CD95 expression in CD4+ T lymphocytes treated with recombinant human erythropoietin. AB - Erythropoietin receptor (EPO-R) appears on the cell surface in the early stages of erythropoiesis. It also has been found on human T and B lymphocytes and monocytes suggesting that EPO could directly influence these cells. Moreover, earlier reports have shown that treatment with recombinant human (rh) EPO in chronic renal failure (CRF) patients improves interleukin-2 production and restores CD4+ T lymphocyte functions. We decided to investigate possibility of direct action of rhEPO on these cells in vitro by phosphorylated signal transducer and activator of transcription 5 (pSTAT5) detection and changes in CD95 antigen expression observation. Flow cytometry was used for detection of pSTAT5 and CD95 expression in CD4+ T lymphocytes treated with rhEPO. Our results show that presence of rhEPO in cell culture of lymphocytes stimulated with anti CD3 antibody increases percentage of CD4+ T lymphocytes expressing pSTAT5. Stimulating effect of rhEPO was dose dependent. RhEPO presence also increases CD95 expression on these cells but still activated T lymphocytes are resistant to CD95-mediated apoptosis. These observations show that EPO is able to directly influence CD4+ T lymphocytes' signaling pathways. PMID- 21619452 TI - Cyclic AMP-induced p53 destabilization is independent of EPAC in pre-B acute lymphoblastic leukemia cells in vitro. AB - CONTEXT: Activation of the tumor suppressor protein p53 facilitates the cellular response to genotoxic stress. Thus, releasing the wild-type p53 from indirect suppression would be crucial to successful killing of cancer cells by DNA damaging therapeutic agents. OBJECTIVE: The aim of this study was to investigate the inhibitory role of cyclic adenosine monophosphate (cAMP) levels on p53 protein in acute lymphoblastic leukemia (ALL) cells. More importantly, we were interested to show through which receptor cAMP acts to promote p53 degradation. MATERIALS AND METHODS: In cell cultures, we investigated the effects of forskolin/3-isobutyl-1-methylxanthine (IBMX) on stimulated p53 of ALL cell lines. Western blotting analysis was performed to detect the expression of p53, phospho p53, acetylated-p53, phospho-cAMP response element-binding protein (CREB), and Mdm2 proteins. Flow cytometry was applied to analyze apoptosis. The gene expression of p53 and its target genes was examined by real-time polymerase chain reaction. RESULTS: We show that elevation of cAMP levels in ALL cells exposed to DNA damage attenuates p53 accumulation. Inhibition of proteosome function with MG 132 reversed the inhibitory effect of cAMP on p53. However, targeting the p53 Mdm2 interaction did not rescue accumulated p53 from the destabilizing signal of cAMP. The specific agonist of the cAMP receptor exchange protein activated by cAMP had no effect on p53 expression in doxorubicin-treated NALM-6 cells, whereas PKA activators decreased p53 accumulation. DISCUSSION AND CONCLUSION: Our studies demonstrate that cAMP-PKA pathway regulates the sensitivity toward DNA-damaging agents via inhibition of a p53-dependent pathway in B-cell precursor ALL (BCP ALL) cells. PMID- 21619453 TI - Rare actinomycetes: a potential storehouse for novel antibiotics. AB - New antimicrobial agents are desperately needed to combat the increasing number of antibiotic resistant strains of pathogenic microorganisms. Natural products remain the most propitious source of novel antibiotics. It is widely accepted that actinobacteria are prolific producers of natural bioactive compounds. We argue that the likelihood of discovering a new compound having a novel chemical structure can be increased with intensive efforts in isolating and screening rare genera of microorganisms. Screening rare actinomycetes and their previously under represented genera from unexplored environments in natural product screening collections is one way of achieving this. Rare actinomycetes are usually regarded as the actinomycete strains whose isolation frequency is much lower than that of the streptomycete strains isolated by conventional methods. Many natural environments are still either unexplored or under-explored and thus, can be considered as a prolific resource for the isolation of less exploited microorganisms. More and different ecological niches need to be studied as sources of a greater diversity of novel microorganisms. In this review, we wish to update our understanding of the potential of the rare actinomycetes by focusing on the ways and means of enhancing their bio-discovery potential. PMID- 21619454 TI - Effects of Teucrium polium spp. capitatum flavonoids on the lipid and carbohydrate metabolism in rats. AB - CONTEXT: The main objective of the study was to investigate the biochemical mechanism of the antidiabetic activities of the dry extracts of Teucrium polium L. ssp. capitatum (L.) Arcangeli (Lamiaceae), from Republic of Macedonia, traditionally used to treat diabetes. MATERIALS AND METHODS: Aerial parts of the plant were extracted in alcohol and freeze- or spray-dried, analyzed by high performance liquid chromatography (HPLC) and examined for insulinotropic effect in INS-1E cells in vitro. Their effect on blood glucose, lipids and carbohydrate related enzymes was tested in normo- and streptozotocin hyperglycemic rats. RESULTS AND DISCUSSION: HPLC analyses revealed several flavonoids: luteolin, apigenin, cirsiliol, diosmetin, cirsimaritin and cirsilineol as both free aglycons and glycosides. The extract and mixture of commercial flavonoids showed a distinct insulinotropic effect on INS-1E cells at 500 ug/ml. Intragastric (i.g.) administration of identical doses of the extract (125 mg/kg) in both normo and hyperglycemic rats was more efficient in lowering the blood glucose than intraperitoneal injection (35% vs. 24% reduction) with highest effect (50% reduction) 8 h after administration. After 10 days of treatment, the magnitude of the effect was comparable to i.g. administration of 2.5 mg/kg of glibenclamide (38% reduction). No effect was seen on blood lipid profiles. In OGTT (oral glucose tolerance test), the extract lowered blood glucose levels by ~35%. The treatment reduced hepatic glycogen and tended to normalize the activity of gluconeogenic enzymes. CONCLUSION: The results demonstrate that examined plant extracts contain flavonoids with insulinotropic and antihyperglycemic effects. PMID- 21619455 TI - Economic burden of resected squamous cell carcinoma of the head and neck in a US managed-care population. AB - BACKGROUND: Squamous cell carcinoma of the head and neck (SCCHN) places a high burden on society and poses complex challenges to healthcare providers. METHODS: Retrospective claims-based analysis of commercially insured patients identified between 01-31-04 and 12-31-07 with diagnostic evidence of cancer of the lip, tongue, oral cavity, pharynx, or larynx who underwent surgical resection during identification period. Outcomes included treatment patterns, healthcare utilization, and costs. All study variables were analyzed descriptively. RESULTS: Among the 1104 patients in the final study sample, 71.9% were male, with mean age 56.6 years. On average, patients were followed for 830 days (range of mean days: 805 for lip or tongue cancer to 847 for pharyngeal cancer). About half received radiation therapy during follow-up, whereas only 16.2% received chemotherapy. Patients with pharyngeal cancer were most likely to undergo chemotherapy. After their index surgery, 57.9% of patients had >=1 inpatient stay, 44.9% had >=1 ER visit, and all had >=1 ambulatory visit. The percentage with >=1 inpatient stay post-index was highest among patients with pharyngeal cancer (73.0%) and lowest in the laryngeal cancer cohort (49.5%). Mean number of hospitalized days, ER visits, and ambulatory visits was 0.45, 0.69, and 27.4, respectively, per-patient per-year. Overall, patients incurred ~$94 million in cost following index surgery ($85,000 per-person, on average). Mean total healthcare cost was $34,450 per patient per-year, the bulk of which comprised medical expenses ($32,401). The highest mean healthcare cost was incurred by the pharyngeal cancer cohort ($40,214). CONCLUSIONS: Patients with resected SCCHN incur substantial healthcare costs and have high utilization rates. Results of this analysis are primarily applicable to resected SCCHN in a managed-care setting, and therefore may not be generalizable to the entire US population. Furthermore, disease stage is an important factor impacting outcomes, but these analyses did not stratify patients according to disease stage. PMID- 21619456 TI - Fundamental and functional aspects of mesoscopic architectures with examples in physics, cell biology, and chemistry. AB - How small can a macroscopic object be made without losing its intended function? Obviously, the smallest possible size is determined by the size of an atom, but it is not so obvious how many atoms are required to assemble an object so small, and yet that performs the same function as its macroscopic counterpart. In this review, we are concerned with objects of intermediate nature, lying between the microscopic and the macroscopic world. In physics and chemistry literature, this regime in-between is often called mesoscopic, and is known to bear interesting and counterintuitive features. After a brief introduction to the concept of mesoscopic systems from the perspective of physics, we discuss the functional aspects of mesoscopic architectures in cell biology, and supramolecular chemistry through many examples from the literature. We argue that the biochemistry of the cell is largely regulated by mesoscopic functional architectures; however, the significance of mesoscopic phenomena seems to be quite underappreciated in biological sciences. With this motivation, one of our main purposes here is to emphasize the critical role that mesoscopic structures play in cell biology and biochemistry. PMID- 21619457 TI - Adverse effects of oral nonstimulant psychotropic medications in young children reported to a regional poison center. AB - BACKGROUND: Use of nonstimulant psychotropic medications other than antidepressants in young children is reported to be increasing. The patient safety ramifications of this remain unclear. OBJECTIVES: To evaluate the frequency of calls to a regional poison center reporting adverse drug effects and the level of medical attention required in young children who are receiving oral nonstimulant psychotropic medications. MATERIALS AND METHODS: A retrospective review of 544 267 consecutive human exposure poison center records between 2000 and 2008 was conducted for cases of young children given nonstimulant psychotropic medications with therapeutic intent. RESULTS: A total of 597 cases met criteria for analysis. Drugs involved were 286 risperidone, 133 clonidine, 114 quetiapine, 37 aripiprazole, 43 olanzapine, 29 ziprasidone, and 5 buspirone; two or more were involved in 250 cases. Reasons for exposure included excess dose given unintentionally (61%), wrong medication unintentionally (12%), adverse effects with correct dose (11%), excess dose intentionally (0.6%), therapeutic error by health-care provider (0.5%), and unclear circumstances (15%). Moderate effects (such as dystonic reaction) occurred in 34 patients at their usual dose (53% of 64) and in 15 at unintentionally excessive doses (4% of 361). Emergency department evaluation of 22% of the children resulted in 5% of the total being admitted to a non-intensive care unit (ICU) bed and 2% of all admitted to an ICU bed. CONCLUSION: Dosing errors and adverse effects involving nonstimulant psychotropic medications are cause for concern in young children. Additional information about safety and optimal dosage of these medications is needed to guide appropriate use. PMID- 21619458 TI - Nutritional and sensory profile of two Indian rice varieties with different degrees of polishing. AB - Traditional hand-pounded rice has been replaced today with highly polished white rice in the Asian Indian diets. The study aimed to evaluate the nutritional as well as the sensory differences between the brown (0% polish) and the rice milled to different degrees of polish (2.3, 4.4 and 8.0%). Bapatla and Uma (red pigmented) varieties in both raw and parboiled forms were used. The protein, fat, dietary fibre, gamma-oryzanol, polyphenols, vitamin E, total antioxidant activity and free radical scavenging abilities of the brown rice decreased while the available carbohydrates increased with polishing. Sensory attributes of the cooked rice samples (whiteness, grain intactness, fluffiness, firmness, stickiness, chewiness and the cooked rice aroma) were evaluated by trained panelists. Scores for branny taste and chewiness decreased with polishing. On the whole, brown rice of both the varieties was readily accepted by the well-informed sensory trained panelists. PMID- 21619459 TI - Influence of combinations of promoter and inhibitor on the bioaccessibility of iron and zinc from food grains. AB - Inherent phytic acid and tannins interfere with bioavailability of iron and zinc from plant-based foods. Food acidulants, beta-carotene-rich vegetables and Allium spices are understood to promote mineral bioaccessibility (an estimate of bioavailability using in vitro method) from food grains. In this study, we have verified whether these promoters would counter negative effects of phytate and tannin on bioaccessibility of iron and zinc from grains. Combinations of promoters - amchur, carrot and onion with phytic acid and tannin exogenously added individually were examined for their influence on iron and zinc bioaccessibility from the food grain. Effect of these promoters was generally dominant in the presence of phytic acid or tannic acid. The negative effect of the inhibitor was not only annulled, but also the positive influence of the promoter was fully retained. This information helps to evolve diet-based strategy to maximize mineral bioavailability and prevent deficiency situations prevalent in population dependent on plant foods. PMID- 21619460 TI - Mild head injury and sympathetic arousal: investigating relationships with decision-making and neuropsychological performance in university students. AB - SUMMARY: The purpose of this study was to examine the relationships between neuropsychological performance, physiological arousal and decision-making in university students who have or have not reported a history of mild head injury (MHI). METHODS: Forty-four students, 18 (41%) reporting a history of MHI, performed a design fluency task and the Iowa Gambling Task (IGT) while electrodermal activity (EDA) was recorded. RESULTS: General cognitive ability and overall choice outcomes did not differ between groups. However, self-reported MHI severity predicted decision-making performance such that the greater the neural indices of trauma, the more disadvantageous the choices made by participants. As expected, both groups exhibited similar base levels of autonomic arousal and physiological responses to reward and punishment outcomes; however, those reporting MHI produced significantly lower levels of EDA during the anticipatory stages of decision-making. CONCLUSIONS: Overall, these findings encourage the acceptance of head injury as being on a continuum of brain injury severity, as MHI can emulate neurophysiological and neuropsychological features of more traumatic cases and may be impacting mechanisms which sustain adaptive social decision-making. PMID- 21619461 TI - Public knowledge of 'concussion' and the different terminology used to communicate about mild traumatic brain injury (MTBI). AB - PRIMARY OBJECTIVE: The primary objective was to investigate the public's general knowledge about concussion (mild traumatic brain injury) and to examine whether terminology influenced attributions made about individuals who experience concussion. DESIGN: A random selection of the community was polled to identify public understanding of concussion. To encourage candid responses, a self-report survey method was used. METHODS AND PROCEDURES: One hundred and three members of the general public were asked to indicate if they associated specific attributes with brain injury or head injury, depending on the randomly assigned questionnaire they completed. Participants also completed a questionnaire about their knowledge of concussion and were asked to indicate whether they or someone they knew had experienced an injury to the head. MAIN OUTCOMES AND RESULTS: Participants incorrectly evaluated 20% (2/10) of statements regarding concussion knowledge and were relatively uncertain about another 20% (2/10) of statements. Negative attributes were associated more with brain than head injury, although those with prior experience made more positive attributions than those without. Fifty-nine per cent of participants who had experienced a concussion stated they had no brain/head injury. CONCLUSIONS: Overall, public knowledge about concussion and different terminology associated with this injury type is substantially inaccurate. More accurate information is required to increase understanding. PMID- 21619462 TI - The role of adiponectin (ADIPOQ) gene polymorphisms in the susceptibility and prognosis of non-small cell lung cancer. AB - To study the role of the adiponectin (ADIPOQ) gene single-nucleotide polymorphism (SNP) in the susceptibility and prognosis for non-small cell lung cancer (NSCLC), we recruited 344 patients with NSCLC, of which 141 had undergone surgical resection and post-surgery follow up. For controls, there were 264 healthy volunteers for the control group, matched in age and sex with the NSCLC patients. Genotyping of SNPs in the ADIPOQ gene, namely, rs266729 (11365C>G); rs822395 (4034A>C); rs822396 (3964A>G); rs2241766 (+45T>G) were performed. Of all SNPs in the ADIPOQ gene, only the TT genotype and T allele frequency of the rs2241766 were more prevalent in NSCLC subjects than in controls. The TT genotype of rs2241766 was significantly associated with susceptibility to NSCLC before and after adjustment for age, sex, body mass index, and smoking status. In the survival analyses of subjects receiving surgical resection, only the SNPs of rs2241766 were significantly related to overall survival of NSCLC. Our results suggest that the SNP rs2241766 of the ADIPOQ gene may determine both susceptibility to NSCLC, and the prognosis for those who underwent surgical treatment. PMID- 21619463 TI - Commutability of the ERM-DA470k reference material for two assays measuring serum albumin using immunochemical principles. PMID- 21619464 TI - Genetics of infectious diseases: hidden etiologies and common pathways. AB - Since the completion of the human genome sequence, the study of common genetic polymorphisms in complex human diseases has become a main activity of human genetics. Employing genome-wide association studies, hundreds of modest genetic risk factors have been identified. In infectious diseases the identification of common risk factors has been varied and as in other common diseases it seems likely that important genetic risk factors remain to be discovered. Nevertheless, the identification of disease-specific genetic risk factors revealed an unexpected overlap in susceptibility genes of diverse inflammatory and infectious diseases. Analysis of the multi-disease susceptibility genes has allowed the definition of shared key pathways of inflammatory dysregulation and suggested unexpected infectious etiologies for other "non-infectious" common diseases. PMID- 21619465 TI - Towards more complete specifications for acceptable analytical performance - a plea for error grid analysis. AB - Abstract We examine limitations of common analytical performance specifications for quantitative assays. Specifications can be either clinical or regulatory. Problems with current specifications include specifying limits for only 95% of the results, having only one set of limits that demarcate no harm from minor harm, using incomplete models for total error, not accounting for the potential of user error, and not supplying sufficient protocol requirements. Error grids are recommended to address these problems as error grids account for 100% of the data and stratify errors into different severity categories. Total error estimation from a method comparison can be used to estimate the inner region of an error grid, but the outer region needs to be addressed using risk management techniques. The risk management steps, foreign to many in laboratory medicine, are outlined. PMID- 21619466 TI - Reference intervals and age and gender dependency for arterial blood gases and electrolytes in adults. AB - BACKGROUND: Although results from blood gas analyzers are frequently used in clinical work surprisingly few and small studies have examined reference intervals for arterial blood gases and acid-base status. We have established reference values based on a large group of healthy people with a wide age distribution. METHODS: A group of medical students (n=182) aged 20-32 years old and a group of health professionals aged 21-76 years were used in this study. Arterial samples were analyzed on the blood gas analyzer ABL from Radiometer(TM). Age and gender dependency was examined for all analytes and reference intervals were calculated non-parametrically. RESULTS: Females had significantly higher pH and lower PaCO(2) (partial pressure of carbon dioxide in an arterial sample), base excess (BE, standard, extra cellular fluid), plasma standard and actual HCO(3), when compared to males (p<0.01). However, the differences were minor and common reference intervals were therefore also determined, generally at the same level as previously published. The lactate values were similar among the genders but with a high upper limit of 2.5 mmol/L. The non-smoker group of females and males had similar PaO(2) values (partial pressure of oxygen in an arterial sample). However, an age dependent effect was found and PaO(2) decreased by 0.29 kPa per decade (confidence interval of slope -0.11 to -0.47 kPa). Electrolytes and anion gap results depicted smaller differences from previous published reference intervals for sodium (136-141 mmol/L) and anion gap (10-16 mmol/L, with potassium included or 6-12 mmol/L without potassium). CONCLUSIONS: Reference intervals for analytes on modern blood gas analyzers were established on a large group of healthy people. Gender and age dependency is generally without clinical importance, except for a lower PaCO(2) in women and a decreasing PaO(2) with higher age. PMID- 21619467 TI - Non-invasive assessment of cardiac output during mechanical ventilation - a novel approach using an inert gas rebreathing method. AB - Measurement of cardiac output (CO) is of importance in the diagnostic of critically ill patients. The invasive approach of thermodilution (TD) via pulmonary artery catheter is clinically widely used. A new non-invasive technique of inert gas rebreathing (IGR) shows a good correlation with TD measurements in spontaneously breathing individuals. For the first time, we investigated whether IGR can also be applied to sedated and mechanically ventilated subjects with a clinical point of care device. CO data from IGR were compared with TD in six healthy mongrel dogs. Data sampling was repeated under baseline conditions (rest) and under stress challenge by applying 10 MUg/kg/min of dobutamine intravenously. Switching from mechanical ventilation to IGR, as well as the rebreathing procedures, were carried out manually. Cardiac output data from IGR and TD correlated with a coefficient of r=0.90 (95% confidence interval [0.81; 0.95]). The Bland-Altman analysis showed a bias of 0.46 l/min for the IGR CO measurements. Ninety-five percent of all differences fall in the interval [-1.03; 1.95], being the limit of the +/- 1.96 standard deviation lines. IGR is a new approach for non-invasive cardiac output measurement in mechanically ventilated individuals, but requires further investigation for clinical use. PMID- 21619468 TI - Intranasal delivery of therapeutic proteins for neurological diseases. AB - INTRODUCTION: Among the range of therapeutic protein candidates for new generation treatments of neurological diseases, neurotrophic factors and recombinant antibodies hold the greatest potential. However, major difficulties in their safe and effective delivery to the brain severely limit these applications. The BBB restricts the exchange of proteins between the plasma and the CNS. Moreover, therapeutic proteins often need to be selectively targeted to the brain, while minimizing their biodistribution to systemic compartments, to avoid peripheral side effects. The intranasal delivery of proteins has recently emerged as a non-invasive, safe and effective method to target proteins to the CNS, bypassing the BBB and minimizing systemic exposure. AREAS COVERED: We critically summarize the main experimental and mechanistic facts about the simple and non-invasive nasal delivery approach, which provides a promising strategy and a potential solution for the severe unmet medical need of safely and effectively delivering protein therapeutics to the brain. EXPERT OPINION: The intranasal route for the effective delivery of recombinant therapeutic proteins represents an emerging and promising non-invasive strategy. Future studies will achieve a detailed understanding of pharmacokinetic and mechanisms of delivery to optimize formulations and fully exploit the nose-to-brain interface in order to deliver proteins for the treatment of neurological diseases. This expanding research area will most likely produce exciting results in the near future towards new therapeutical approaches for the CNS. PMID- 21619469 TI - Physical hydrogels with self-assembled nanostructures as drug delivery systems. AB - INTRODUCTION: As an essential complement to chemically crosslinked hydrogels, drug delivery systems based on physical hydrogels with self-assembled nanostructures are gaining increasing attention, owing to potential advantages of reduced toxicity, convenience of in situ gel formation, stimuli-responsiveness, reversible sol-gel transition, and improved drug loading and delivery profiles. AREAS COVERED: In this review, drug delivery systems based on physical hydrogels are discussed according to their self-assembled nanostructures, such as micelles, layer-by-layer constructs, supramolecular inclusion complexes, polyelectrolyte complexes and crystalline structures. The driving forces of the self-assembly include hydrophobic interaction, hydrogen bonding, electrostatic interaction, pi pi stacking and weak van der Waals forces. Stimuli-responsive properties of physical hydrogels, including thermo- and pH-sensitivity, are considered with particular focus on self-assembled nanostructures. EXPERT OPINION: Fabricating self-assembled nanostructures in drug delivery hydrogels, via physical interactions between polymer-polymer and polymer-drug, requires accurately controlled macro- or small molecular architecture and a comprehensive knowledge of the physicochemical properties of the therapeutics. A variety of nanostructures within hydrogels, with which payloads may interact, provide useful means to stabilize the drug form and control its release kinetics. PMID- 21619470 TI - P2X7 receptor antagonism in the treatment of cancers. AB - ATP-gated P2X7 receptors (P2X7) make a unique family of extracellular ATP activated plasma membrane ion channels expressed in haematopoietic and epithelial cells. They have been extensively studied in immune cells where their activation leads to the rapid release of pro-inflammatory cytokines and the initiation of the inflammatory cascade. As such, P2X7 represent a pharmaceutical target for the treatment of inflammatory diseases. Recently, P2X7 expression has been found in diverse tumours and has been suggested as a potential cancer cell biomarker. On ATP stimulation, tumour cells can use P2X7 signalling in different scenarios: i) as a reaction to this death-related signal, they can downregulate P2X7 to avoid apoptosis or ii) as a cancer-promoting signal to survive and enhance invasion of new niches. The high levels of extracellular ATP found in tumours could represent a stressful stimulus for cancer cells by initiating P2X7-driven cell death. Therefore, the increased P2X7-dependent invasiveness of cancer cells could be an escape strategy to flee the noxious high level of ATP. The use of specific P2X7 antagonists could be a new alternative way to reduce the development of cancer metastases and improve the efficacy of conventional treatments. PMID- 21619471 TI - Antifolate agents: a patent review (2006 - 2010). AB - INTRODUCTION: For > 50 years, drugs targeting the folate pathway have significantly impacted disease treatment as anticancer, antimicrobial and immunomodulatory agents. The discovery of novel antifolate agents with improved properties and superior activities remains an attractive strategy, both in academia and the pharmaceutical industry. AREAS COVERED: This review surveys the patent literature from 2006 to 2010 for small molecule inhibitors of enzymatic targets in the folate biosynthetic pathway. EXPERT OPINION: The pursuit of antifolates as anticancer and antimicrobial agents continues to be an active area of research. New patent disclosures reveal novel antifolate scaffolds, antifolates with improved drug-like properties and new strategies to effectively target cancer cells. The continued use of high resolution structural information has guided the discovery of several compounds. Owing to the need for high levels of potency and selectivity, especially in targeting pathogenic species, the use of high resolution crystal structures remains an important tool to guide the design of novel antifolates. Interestingly, the patents disclosing novel compounds were ones where X-ray crystallography was an integral component of the design process. Finally, a variety of new structures have been reported that may play an important role in the future development of therapeutic antifolates. PMID- 21619472 TI - The effect and clinical consequences of hypoxia on cytochrome P450, membrane carrier proteins activity and expression. AB - INTRODUCTION: Chronic pulmonary disease and heart failure reduce drug clearance and consequently enhance adverse drug reactions. The mechanisms of action underlying the regulation of cytochrome P450 (CYP) isoforms and membrane carrier proteins by hypoxia, and the clinical consequences of the regulation of CYP by hypoxia, alone or combined with other conditions have been elucidated in the last decades. Overall, a reduced drug clearance appears to be associated with hypoxemia. AREAS COVERED: In this review, the mechanisms of action underlying hypoxia-induced regulation of CYP enzymes are discussed. The authors also revise the effects of hypoxia on serum mediators, signal transduction pathways, orphan nuclear receptors, transcription factors and post-transcriptional mechanisms regulating CYP and membrane carrier proteins expression. Additionally, the paper also discusses the clinical repercussions of hypoxia-induced changes in CYP and membrane carrier proteins activity. EXPERT OPINION: Acute systemic hypoxia down regulates selected CYP isoforms and up-regulates CYP3A4 and P-glycoprotein, changing the metabolic clearance of drugs and endogenous compounds biotransformed by these isoforms as well as the kinetics. In patients with acute hypoxia, the dosage of drugs, biotransformed by CYP isoforms, may need to be adjusted. Tissue hypoxia enhances the expression of efflux membrane carrier proteins, increasing the probability of drug resistance. PMID- 21619473 TI - Evaluation of two automated chemiluminescence immunoassays, the LIAISON Treponema Screen and the ARCHITECT Syphilis TP, and the Treponema pallidum particle agglutination test for laboratory diagnosis of syphilis. AB - BACKGROUND: Automated Treponema pallidum-specific chemi-luminescence immunoassays (CLIA) run on random-access analyzers allow for rapid diagnosis of syphilis infection. METHODS: We evaluated the LIAISON Treponema Screen (LIA) and the ARCHITECT Syphilis TP (ARCH) in comparison to the Treponema pallidum particle agglutination (TPPA) test, as a screening test for syphilis. We performed a prospective study using 577 sera submitted for diagnosis of syphilis, including 318 samples from pregnant women. In addition, 42 stored sera from 32 patients with clinically and serologically characterized syphilis infection were investigated. RESULTS: In the prospective study, the sensitivity and specificity of LIA, ARCH, and TPPA were 100% (18/18), 100% (17/17), and 100% (18/18), and 100% (558/558), 99.8% (552/553), and 99.6% (556/558), respectively. In pregnant women, the specificity of LIA and ARCH was 100% (317/317) and of TPPA 99.7% (316/317). One sample from a child with assumed exposure to maternal antitreponemal antibodies was omitted from analysis. LIA, ARCH, and TPPA were also positive in all investigated sera from patients with known syphilis. CONCLUSIONS: Both automated CLIA demonstrated excellent diagnostic sensitivity and specificity when evaluated as a screening test for syphilis under routine conditions of a diagnostic laboratory. Thus, these may be used independently as an alternative to the manual TPPA screen. PMID- 21619474 TI - Current limitations of the Athlete's Biological Passport use in sports. AB - The Athletes Biological Passport (ABP) has received both criticisms and support during this year. In a recent issue of The Lancet, Michael Wozny considered that the use of the ABP makes it more difficult to take banned substances and that it was successfully used against the Italian elite cyclist Franco Pellizotti. After that, Italy's anti-doping tribunal considered that there was not enough evidence to prove manipulation of his own blood profile in Pellizotti's case. However, the UCI appealed to the Court of Arbitration for Sport (CAS) that sanctioned Pellizotti with a suspension of 2 years. Since its implementation, some problems have emerged. From 2010 to date, a large number of reports regarding the stability of the blood variables used to determine the ABP have been published, showing mixed results. This study considers that there is a risk of misinterpreting the physiological variations of the hematological parameters determined by the anti-doping authorities in the ABP. The analytical variability due to exercise training and competitions and/or to different metabolic energy demands, hypoxia treatments, etc. could lead to an increase in false-positives when using the ABP with the dramatic consequences that they might cause in major sports events like the forthcoming London Olympic Games. Moreover, the ABP characteristics, procedures, thresholds, or individual determination of reference ranges, abnormal out-comes, strikes, "how the profile differs from what is expected in clean athletes" should be clearly stated and explained in a new public technical document to avoid misunderstandings and to promote transparency. PMID- 21619475 TI - Limits and pitfalls of Athlete's Biological Passport. AB - The Athlete's Biological Passport (ABP) is an evaluation of hematological parameters, hemoglobin (Hb), reticulocytes (Ret), and their combination in the OFF-score. Recently, the Court of Arbitration for Sport accepted it as a suitable indirect method for detecting blood doping. There are various topics which are not defined and scientifically completely explained in ABP, limiting its effectiveness as evidence and as suspect of blood manipulation. The data source the ABP used for designing a profile is unclear. The variance used for cyclists is not correct. The covariables which should be calculated together with the measures of Hb and Ret are not always considered in the statistical program. The pre-analytical warnings for correct and valid collection, transport, and storage of the specimens are not assured. Quality control of the instruments is not completely assured. Analytical variability is not appropriately considered in the program. The seasonal changes of the hematological parameters, due to training and competitions, are not calculated. Statistical analysis, based on a Bayesian like program, not available to the scientific community, does not follow the classical decision-making approach of medicine and science. The ABP needs of additional evidences and of scientific debate. PMID- 21619476 TI - Performance evaluation of the Vitros((r))3600 immunodiagnostic system for the determination of free thyroid hormones. PMID- 21619477 TI - Improving clinical laboratory efficiency: a time-motion evaluation of the Abbott m2000 RealTime and Roche COBAS AmpliPrep/COBAS TaqMan PCR systems for the simultaneous quantitation of HIV-1 RNA and HCV RNA. AB - BACKGROUND: Diagnostic laboratories need automation that facilitates efficient processing and workflow management to meet today's challenges for expanding services and reducing cost, yet maintaining the highest levels of quality. METHODS: Processing efficiency of two commercially available automated systems for quantifying HIV-1 and HCV RNA, Abbott m2000 system and Roche COBAS Ampliprep/COBAS TaqMan 96 (docked) systems (CAP/CTM), was evaluated in a mid/high throughput workflow laboratory using a representative daily workload of 24 HCV and 72 HIV samples. Three test scenarios were evaluated: A) one run with four batches on the CAP/CTM system, B) two runs on the Abbott m2000 and C) one run using the Abbott m2000 maxCycle feature (maxCycle) for co-processing these assays. Cycle times for processing, throughput and hands-on time were evaluated. RESULTS: Overall processing cycle time was 10.3, 9.1 and 7.6 h for Scenarios A), B) and C), respectively. Total hands-on time for each scenario was, in order, 100.0 (A), 90.3 (B) and 61.4 min (C). CONCLUSIONS: The interface of an automated analyzer to the laboratory workflow, notably system set up for samples and reagents and clean up functions, are as important as the automation capability of the analyzer for the overall impact to processing efficiency and operator hands on time. PMID- 21619478 TI - Reply to Vavrova et al. Clin Chem Lab Med 2011;49:89-92. PMID- 21619479 TI - Agreement between paired blood gas values in samples transported either by a pneumatic system or by human courier. AB - BACKGROUND: Rapid accurate assessment of metabolic derangements is crucial in the critically ill. We evaluated if arterial blood gas (ABG) samples transported through a pneumatic tube system (PTS) agreed with values transported by a human courier. METHODS: In this prospective study of 50-paired ABG samples, the couriered reference ABG was compared with those transported by PTS. Agreement was summarised by the mean difference with 95% limits of agreement (LOA) and Lin's concordance correlation (pc). RESULTS: The mean (+/-SD) time from sampling to analysis was 35.7+/-23.2 (courier) and 38.6+/-22.1 (PTS) minutes. Agreement was good between courier and PTS for pH, PaCO(2), bicarbonate, oxygen saturation and PaO(2) values (pc>0.97). Although the mean difference in PaO(2) values between PTS and courier was small (-0.9 mm Hg) and the agreement was good, individual differences were clinically significant (95% LOA -40.8 to 39.0). For PaO(2) <160 mm Hg, analysis of PTS samples yielded erroneously high PaO(2) values and vice versa for PaO(2)>160 mm Hg compared to manual courier. This suggested exaggerated oxygen movement between the blood sample and air in the PTS. CONCLUSIONS: In this study, analysis of samples transported through the PTS resulted in clinically unacceptable PaO(2) values. Delay in transport and analysis of ABG samples should be avoided and samples transported manually if they cannot be assessed on-site. PMID- 21619480 TI - Gender-related differences in the oxidant state of cells in Fanconi anemia heterozygotes. AB - Abstract Fanconi anemia (FA) is a rare cancer-prone genetic disorder characterized by progressive bone marrow failure, chromosomal instability and redox abnormalities. There is much biochemical and genetic data, which strongly suggest that FA cells experience increased oxidative stress. The present study was designed to elucidate if differences in oxidant state exist between control, idiopathic bone marrow failure (idBMF) and FA cells, and to analyze oxidant state of cells in FA heterozygous carriers as well. The results of the present study confirm an in vivo prooxidant state of FA cells and clearly indicate that FA patients can be distinguished from idBMF patients based on the oxidant state of cells. Female carriers of FA mutation also exhibited hallmarks of an in vivo prooxidant state behaving in a similar manner as FA patients. On the other hand, the oxidant state of cells in FA male carriers and idBMF families failed to show any significant difference vs. controls. We demonstrate that the altered oxidant state influences susceptibility of cells to apoptosis in both FA patients and female carriers. The results highlight the need for further research of the possible role of mitochondrial inheritance in the pathogenesis of FA. PMID- 21619481 TI - Cooperative and independent activities of Sgt2 and Get5 in the targeting of tail anchored proteins. AB - Abstract TPR proteins modulate the activity of molecular chaperones. Here, we describe the S. cerevisiae TPR protein Sgt2 as interaction partner of Ssa1 and Hsp104 and as a component of the GET pathway by interacting with Get5. The GET pathway mediates the sorting of tail-anchored (TA) proteins, harboring a C terminal trans-membrane segment, to the ER membrane. S. cerevisiae sgt2Delta cells show partial defects in TA protein sorting. Sgt2 activity in vivo relies on its N- and C-terminal domains, whereas the central TPR domain and thus chaperone interactions are dispensable. We show that TA protein sorting defects are more severe in sgt2Delta get5Delta mutants compared to single knockouts. Furthermore, overproduction of Sgt2 becomes toxic to get3Delta but not to get5Delta cells. Together, these findings indicate an additional, Get5-independent role of Sgt2 in TA protein sorting, pointing to parallel pathways of substrate delivery to Get3. PMID- 21619482 TI - Improved detectability of small-bowel lesions via capsule endoscopy with computed virtual chromoendoscopy: a pilot study. AB - OBJECTIVE: Real-time video capsule endoscopy (CE) with flexible spectral imaging color enhancement (FICE) improves visibility of small-bowel lesions. This article aims to clarify whether CE-FICE also improves detectability of small-bowel lesions. PATIENTS AND METHODS: A total of 55 patients who underwent CE at Hiroshima University Hospital during the period November 2009 through March 2010 were enrolled in the study. Five patients were excluded from the study because residues and transit delays prevented sufficient evaluation. Thus, 50 patients participated. Two experienced endoscopists (each having interpreted more than 50 capsule videos) analyzed the images. One interpreted conventional capsule videos; the other, blinded to interpretation of the conventional images, interpreted CE FICE images obtained at settings 1-3 (setting 1: red 595 nm, green 540 nm, blue 535 nm; setting 2: red 420 nm, green 520 nm, blue 530 nm; setting 3: red 595 nm, green 570 nm, blue 415 nm). Lesions were classified as angioectasia, erosion, ulceration, or tumor. Detectability was compared between the two modalities. Time taken to interpret the capsule videos was also determined. RESULTS: Seventeen angioectasias were identified by conventional CE; 48 were detected by CE-FICE at setting 1, 45 at setting 2, and 24 at setting 3, with significant differences at settings 1 and 2 (p = 0.0003, p < 0.0001, respectively). Detection of erosion, ulceration, and tumor did not differ statistically between conventional CE and CE FICE, nor did interpretation time (conventional CE 36 +/- 6.9 min; CE-FICE setting 1, 36 +/- 6.4 min; setting 2, 38 +/- 5.8 min; setting 3, 35 +/- 6.7 min). CONCLUSIONS: CE-FICE is superior in the lesion detection in comparison with conventional CE and improves detection of angioectasia. PMID- 21619483 TI - Evaluation of disease activity in IBD at the time of diagnosis by the use of clinical, biochemical, and fecal markers. AB - OBJECTIVE: The present population based adult cohort was part of a new prospective study of patients with inflammatory bowel disease (IBD) in South Eastern Norway, the Inflammatory Bowel South-Eastern Norway II study, investigating disease characteristics in an attempt to improve our knowledge regarding factors related to early clinical phenotype and disease activity. MATERIAL AND METHODS: Patients suspected to have IBD on the basis of predefined symptoms, including abdominal pain, diarrhea, and/or blood in stool for more than 10 days were examined at the local hospital. Colonoscopy with biopsies was performed and blood and stool samples were taken. RESULTS: In ulcerative colitis (UC) patients, the median Simple Clinical Colitis Activity Index (SCCAI) was 4 (range 0-10) in mild and 6 (range 0-14) in patients with moderate or severe endoscopic activity of inflammation (p = 0.002). The calprotectin concentration in feces was significantly related to the SCCAI (p = 0.034) and the Mayo endoscopic subscore (p = 0.031). There was a significant association between the C-reactive protein (CRP) value, leucocytes and thrombocytes and the SCCAI, but only leucocytes were significantly associated with the Mayo endoscopic subscore. In Crohn's disease (CD) patients, there was no statistical significant association between the Harvey-Bradshaw Index (HBI) and the endoscopic grade of mucosal inflammation (p = 0.8). The calprotectin concentration in feces was significantly related to the endoscopic activity score (p = 0.004), but not to the HBI (p = 0.5). HBI was significantly related to the CRP value (p = 0.047) and thrombocytes (p = 0.03). CONCLUSIONS: In UC, both biochemical and fecal markers are related to disease activity and extent of disease, whereas in CD, the fecal calprotectin concentration is a reliable marker of mucosal affection, but not for systemic disease activity. PMID- 21619484 TI - Identification of a redox-modulatory interaction between uncoupling protein 3 and thioredoxin 2 in the mitochondrial intermembrane space. AB - Uncoupling protein 3 (UCP3) is a member of the mitochondrial solute carrier superfamily that is enriched in skeletal muscle and controls mitochondrial reactive oxygen species (ROS) production, but the mechanisms underlying this function are unclear. AIMS: The goal of this work focused on the identification of mechanisms underlying UCP3 functions. RESULTS: Here we report that the N terminal, intermembrane space (IMS)-localized hydrophilic domain of mouse UCP3 interacts with the N-terminal mitochondrial targeting signal of thioredoxin 2 (Trx2), a mitochondrial thiol reductase. Cellular immunoprecipitation and in vitro pull-down assays show that the UCP3-Trx2 complex forms directly, and that the Trx2 N-terminus is both necessary and sufficient to confer UCP3 binding. Mutation studies show that neither a catalytically inactivated Trx2 mutant, nor a mutant Trx2 bearing the N-terminal targeting sequence of cytochrome c oxidase (COXMTS-Trx2) bind UCP3. Biochemical analyses using permeabilized mitochondria, and live cell experiments using bimolecular fluorescence complementation show that the UCP3-Trx2 complex forms specifically in the IMS. Finally, studies in C2C12 myocytes stably overexpressing UCP3 (2.5-fold) and subjected to Trx2 knockdown show that Trx2 is required for the UCP3-dependent mitigation of complex III-driven mitochondrial ROS generation. UCP3 expression was increased in mice fed a high fat diet, leading to increased localization of Trx2 to the IMS. UCP3 overexpression also increased expression of the glucose transporter GLUT4 in a Trx2-dependent fashion. INNOVATION: This is the first report of a mitochondrial protein-protein interaction with UCP3 and the first demonstration that UCP3 binds directly, and in cells and tissues with mitochondrial thioredoxin 2. CONCLUSION: These studies identify a novel UCP3-Trx2 complex, a novel submitochondrial localization of Trx2, and a mechanism underlying UCP3-regulated mitochondrial ROS production. PMID- 21619485 TI - Aminopeptidase N (EC 3.4.11.2) inhibitors (2006 - 2010): a patent review. AB - INTRODUCTION: Aminopeptidase N (APN/CD13) acts as an exopeptidase and has been studied for decades. In recent years, it has been seen not only as a tumor related target but also as a potential functional protein in various other physiological or pathological processes, such as analgesia, virus infection and inflammation. AREAS COVERED: In this review, APN inhibitors in the patents publicized during 2006 - 2010 are introduced. Readers will gain information on the patent inhibitors, including chemical structures, original sources or synthetic methods, biological assays and application potential. EXPERT OPINION: It is difficult to identify compounds that interact with the function not relevant with peptide-hydrolysis of APN in the enzyme activity assay, and such compounds have not been reported in the patents during the past 5 years. The progress of protein-small molecule interaction detecting means, such as surface plasmon resonance, will possibly help develop such compounds for the treatment of relevant diseases or new molecular probes in mechanism investigation. PMID- 21619486 TI - Psychiatric symptomatologies and disorders related to epilepsy and antiepileptic medications. AB - INTRODUCTION: Psychiatric comorbidities (such as depression, anxiety, psychosis, inattention, obsession, personality traits, aggression and suicide) are frequent in patients with epilepsy and have a significant impact on medical management and quality of life. AREAS COVERED: A literature search was performed in MEDLINE for epidemiological, longitudinal, prospective, double-blind clinical trial studies published between 1990 and 2011 using the following words: epilepsy, antiepileptic drugs (AEDs), behavioral/emotional/psychiatric comorbidities, suicide and aggression. In this review, the author discusses: i) the characterization and prevalence of behavioral disturbances associated with epilepsy, ii) variables correlated with behavioral comorbidities which include: psychosocial-, clinical- and treatment-related variables, iii) the complex mechanisms of behavioral comorbidities associated with epilepsy, which include both psychosocial (functional) and organic; the process of epileptogenesis, neuronal plasticity, abnormalities in hypothalamic-pituitary axis and neurotransmitters and pathways are fundamental determinants, iv) the negative psychotropic effects of AEDs and their mechanisms and v) the suggested biopsychosocial model of management (pharmacological and non-pharmacological). EXPERT OPINION: The relationship between psychiatric disorders and epilepsy has relevant therapeutic implications which should be directed towards a comprehensive biopsychosocial approach that focuses on the whole person rather than simply on the disease process. PMID- 21619487 TI - The Therapeutic Target Database: an internet resource for the primary targets of approved, clinical trial and experimental drugs. AB - Increasing numbers of proteins, nucleic acids and other molecular entities have been explored as therapeutic targets. A challenge in drug discovery is to decide which targets to pursue from an increasing pool of potential targets, given the fact that few innovative targets have made it to the approval list each year. Knowledge of existing drug targets (both approved and within clinical trials) is highly useful for facilitating target discovery, selection, exploration and tool development. The Therapeutic Target Database (TTD) has been developed and updated to provide information on 358 successful targets, 251 clinical trial targets and 1254 research targets in addition to 1511 approved drugs, 1118 clinical trials drugs and 2331 experimental drugs linked to their primary targets (3257 drugs with available structure data). This review briefly describes the TTD database and illustrates how its data can be explored for facilitating target and drug searches, the study of the mechanism of multi-target drugs and the development of in silico target discovery tools. PMID- 21619488 TI - Mitochondria and programmed cell death in Parkinson's disease: apoptosis and beyond. AB - Abstract Significance: Activation of mitochondrion-dependent programmed cell death (PCD) pathways is instrumental to the demise of substantia nigra pars compacta dopaminergic neurons in experimental mouse models of Parkinson's disease (PD). Supporting the relevance of these findings for PD, key molecular elements of this pathogenic cascade have also been demonstrated in postmortem brain samples of PD patients. Recent Advances and Critical Issues: Mounting evidence indicates that different morphological types of cell death co-exist in the brain of PD patients, all of which may result from the activation of common upstream PCD pathways. Indeed, contrary to initial views, it is now established that the deleterious effects of PCD pathways are not limited to mitochondrion-mediated caspase-dependent apoptosis but also involve caspase-independent nonapoptotic cell death, including necrosis. This notion may help reconcile the observation of both apoptotic and nonapoptotic dopaminergic cell death in postmortem PD samples. FUTURE DIRECTIONS: Potential neuroprotective strategies for PD should be aimed at targeting both apoptotic and nonapoptotic pathways, all of which may simultaneously occur in PD patients through activation of common upstream PCD pathways involving the mitochondria. Antioxid. Redox Signal. 16, 883-895. PMID- 21619489 TI - Long-term mortality rate in rheumatoid arthritis patients with disease onset in the 1980s. AB - OBJECTIVE: To investigate the mortality rate and possible early predictive factors of mortality after 19-23 years in a cohort of patients with rheumatoid arthritis (RA) followed prospectively from disease onset. PATIENTS AND METHODS: A community-based cohort of 183 patients (63% female) with RA and disease duration < 2 years was recruited 1985-1989. The patients were followed yearly from diagnosis until death or 31 December 2008. Mean age and mean duration of symptoms (range) at diagnosis were 52 (18-78) years and 11 (0-24) months, respectively. Death certificates were obtained from the Swedish Cause of Death Register and causes of death were coded by the International Classification of Diseases (ICD 10). Death rates of RA patients were compared to those of age- and sex-matched controls. Possible predictors of mortality were analysed using a Cox regression model. RESULTS: By 31 December 2008, 69 patients (37 women and 32 men) had died. The standardized mortality ratio (SMR) was 1.23 [95% confidence interval (CI) 0.97-1.55] and p < 0.09. Older age, male sex, smoking, and the presence of cardiovascular disease (CVD) at RA diagnosis were identified as early predictors of mortality. CVD was the most common cause of death (46%), followed by malignancies (29%) and infections (13%). RA was not stated as the direct cause of death in any patient and was mentioned among underlying causes in only 16/69 (23%) patients. CONCLUSION: Mortality rate after 19-23 years of disease duration in this cohort of patients with disease onset in the 1980s was not significantly increased compared to age- and sex-matched controls. No RA disease-related factor predicted mortality. PMID- 21619490 TI - Cystatin C concentration is correlated with disease activity in rheumatoid arthritis patients. AB - OBJECTIVES: Rheumatoid arthritis (RA) is a systemic, inflammatory disease. Renal involvement worsens the course of RA and increases mortality. It is suggested that chronic inflammatory processes may contribute to renal impairment. The aim of this study was to investigate the impact of chronic inflammation and RA activity on glomerular filtration rate (GFR). METHODS: The study population consisted of 140 RA patients. High disease activity was observed in 42 patients (30%), and long-term RA (duration >= 10 years) in 64 (45.7%). Measures of renal function included: serum cystatin C, serum creatinine (SCr), and creatinine-based estimated GFR (eGFR) calculated by Cockcroft and Gault (CG) and Modification of Diet in Renal Disease (MDRD) formulas. RESULTS: The mean (SD) cystatin C concentration was 0.77 (0.2) mg/L, SCr 0.71 (0.23) mg/dL, eGFR(CG) 110.5 (37.8) mL/min/1.73 m2, and eGFR(MDRD) 109.5 (34.5) mL/min/1.73 m2. Cystatin C levels correlated positively with creatinine, and negatively with eGFR(CG) and eGFR(MDRD). Cystatin C concentration was significantly higher in patients with high disease activity, long-term RA, and hypertension, and in males. Patients currently being treated with biologics had non-significantly lower cystatin C levels than those treated with conventional modifying drugs. Cystatin C levels were significantly associated with markers of clinical, functional disease activity, and markers of inflammation. By contrast, there were no such correlations with other parameters of renal function. CONCLUSIONS: In patients with RA, cystatin C may be not only an indicator of GFR but also a marker of intensity of chronic inflammatory processes. PMID- 21619491 TI - Intra-articular corticoid injection induces circulating glucocorticoid bioactivity and systemic immune activation in juvenile idiopathic arthritis. AB - OBJECTIVE: To study the systemic effects of intra-articular (IA) glucocorticoid (GC) injections in juvenile idiopathic arthritis (JIA). METHODS: The study group comprised 21 JIA patients being treated with IA methylprednisolone [MP (n = 15) or MP plus triamcinolone hexacetonide (THA) (n = 6)] prescribed on clinical indications. The systemic effect of MP was assessed by measuring circulating glucocorticoid bioactivity (GBA) with a recombinant cell transactivation assay 7 and 24 h after the IA injections, and after 2 months. The systemic immunological responses were studied with a novel assay for testing patient serum-induced changes in the secretion of interferon (IFN)-gamma and interleukin (IL)-5 from target cells. RESULTS: Administration of IA GC induced serum GBA (p = 0.001) and suppressed circulating cortisol levels (p = 0.002) 7 h after the injection. Serum withdrawn 24 h after the IA injection induced less IL-5 secretion from mitogen activated target cells when compared with pre-treatment sera (p = 0.036). This decrease in target cell T helper (Th)2 response (IL-5) was MP dose related (r = 0.550, p = 0.018). High IL-5 secretion from target cells prior to the IA injections was associated with good clinical outcome at 2 months, seen as a low number of active (p = 0.044) and restricted joints (p = 0.049). CONCLUSION: IA GC injections have systemic effects that are reflected in the serum as an immediate elevation of GBA, a decrease of endogenous cortisol as well as a suppressive effect of patient serum on target cell IL-5 secretion. These systemic effects may play a role in the attenuation of disease activity. PMID- 21619493 TI - Monitoring human mesenchymal stromal cell differentiation by electrochemical impedance sensing. AB - BACKGROUND AIMS: For their wide mesodermal differentiation potential, mesenchymal stromal/stem cells (MSC) are attractive candidates for tissue engineering. However, standardized quality control assays monitoring differentiation that are non-invasive and continuous over time are lacking. METHODS: We employed a non invasive assay, using two different systems, to discriminate osteogenic and adipogenic differentiation of MSC by monitoring impedance. Fibroblasts and keratinocytes served as non-specific controls. Impedance profiles were recorded comparing MSC from bone marrow and adipose tissue, either non-induced or induced for osteogenesis or adipogenesis, for 5-14 days, and correlated with differentiation markers assessed by reverse transcription-quantitative polymerase chain reaction and Western blot. Additionally, differentiation modulating effects of extracellular matrix components were analyzed. RESULTS: Adhesion and growth related impedance profiles of non-induced MSC roughly resembled those of fibroblasts, whereas keratinocytes differed significantly. Distinct from that, osteogenic induction of MSC revealed initially rapid and continuously rising impedance, corresponding to mineralized calcium matrix formation. Conversely, adipogenic induction caused shallower initial slopes and eventually declining profiles, corresponding to more compact, adipocyte-like cells with numerous lipid vacuoles. Pre-coating with either collagen type I or IV apparently favored osteogenesis and fibronectin adipogenesis. Impedance recordings correlated well with the extent of differentiation evaluated by histochemical staining and protein and gene expression. CONCLUSIONS: Overall, our data demonstrate that impedance profiling offers a basis for standardized real-time, non-invasive high throughput screening of MSC properties. It enables further testing of the influence of diffusible factors or extracellular matrix composites on MSC differentiation or maintenance of stemness, thus substantiating therapeutic application. PMID- 21619495 TI - A novel SDHD mutation associated with neck paraganglioma. AB - The aim of this study was to describe a previously unreported mutation in the SDHD gene, which has been linked to familial paraganglioma. Clinical data were collected from all members of the family, which had four siblings affected with paraganglioma. For the index patient, genomic DNA extraction from whole blood was performed using the High Pure PCR Template Preparation kit. The nucleotide sequence in the index patient revealed a deletion in the SDHD gene, c.165_169 + 14del. The loss of nucleotides in the DNA led to production of an anomalous protein. RNA analysis showed the absence of exon 2 in the sequence that corresponded to the mRNA from the index case. Genetic testing of this deletion was extended to the symptomatic and asymptomatic brothers and sisters of the index patient and other family members at risk. The deletion was detected in both symptomatic brothers, in accordance with their phenotype, but not in the asymptomatic sister. In the other asymptomatic brother (II.7) the deletion was detected and magnetic resonance angiography revealed the vascular characteristics of two tumors in both carotid bifurcations. Thus, we report a novel punctual mutation in the SDHD gene, which is related to familial paraganglioma: the deletion was c.165_169 + 14del. PMID- 21619494 TI - Comparative activity of tigecycline and tetracycline on Gram-negative and Gram positive bacteria revealed by a multicentre study in four North European countries. AB - BACKGROUND: This study involves a multicentre surveillance of tigecycline and tetracycline activity against Gram-negative and Gram-positive bacteria from primary care centres (PCCs), general hospital wards (GHWs) and intensive care units (ICUs) in Denmark (n = 9), Finland (n = 10), Norway (n = 7) and Sweden (n = 19). METHODS: The hospitals were each asked to test 30 consecutive Gram-positive and 30 Gram-negative clinical isolates. Supportive information accompanying each isolate included the study centre, ward level (PCC, GHW, or ICU), patient identification and source of the isolate. Minimum inhibitory concentrations (MICs) for tetracycline and tigecycline were determined with the Etest. RESULTS: The isolates collected comprised 1610 Gram-negative and 1767 Gram-positive clinical isolates. The study showed low rates of non-susceptibility (intermediate (I) and resistant (R)) to tigecycline: <1% in Escherichia coli, though other Enterobacteriaceae showed higher rates (Enterobacter cloacae (7%), Klebsiella pneumoniae (9%) and Serratia spp. (23%)). The overall non-susceptibility rate for tigecycline in Enterobacteriaceae with species-related breakpoints for tigecycline was 6% (4% excluding Serratia spp.). The activity of tigecycline against Haemophilus influenzae and Acinetobacter spp. was high with a MIC(50) of 0.25 mg/l and MIC(90) of 1 mg/l. The prevalence of non-susceptibility to tigecycline among Gram-positive bacteria was <1%. The corresponding figure for tetracycline was 14%. The activity of tigecycline against Streptococcus pneumoniae was high with MIC(50) and MIC(90) of 0.125 mg/l. CONCLUSION: Tigecycline showed good overall in vitro activity against Gram-positive and Gram negative isolates, including both tetracycline-susceptible and resistant isolates. Most non-susceptibility to tigecycline among Enterobacteriaceae other than E. coli was I (6%), rather than R (<1%). This indicates a problem setting interpretive species-related tigecycline breakpoints for Enterobacteriaceae other than E. coli. PMID- 21619496 TI - Invasive Myceliophthora thermophila infection mimicking invasive aspergillosis in a neutropenic patient: a new cause of cross-reactivity with the Aspergillus galactomannan serum antigen assay. AB - Myceliophthora thermophila is a thermophilic mould widely found in the environment but rarely responsible for human infections. We describe a case of invasive Myceliophthora thermophila infection mimicking invasive aspergillosis in a neutropenic patient with haematological malignancy. Cross-reactivity with Aspergillus galactomannan assay (GM) was demonstrated by repeated positive results and confirmed by cross-reaction between the fungal isolate and the GM assay. The patient was successfully treated with voriconazole. Potential GM cross reactivity must be considered in future studies including patients categorized as having probable invasive aspergillosis using the GM as the only mycological criterion. PMID- 21619497 TI - Susceptibility of Candida species and various moulds to antimycotic drugs: use of epidemiological cutoff values according to EUCAST and CLSI in an 8-year survey. AB - A collection of 2,834 isolates of Candida spp. and 1,079 isolates of Aspergillus spp. and other moulds that were recovered between 2000 and 2007 in Tyrol, Austria, were examined for their susceptibility to antifungal drugs. The susceptibility of Candida spp. to amphotericin B (AMB), caspofungin (CPF), fluconazole (FLC), and voriconazole (VRC) were studied, while filamentous fungi were tested against AMB, CPF, VRC, itraconazole (ITC), and posaconazole (POS). As EUCAST and CLSI are currently revising their breakpoints for several antifungal agents, epidemiological cutoff values (ECVs) of these two guidelines were used to examine trends in susceptibility. For Candida spp. we noted increases in the percentage of non-wild type isolates which were resistant to CPF, FLC, and VRC. Most noticeably, we observed a change in: C. tropicalis (from 0.9-3.8%) and C. parapsilosis (from 4.0-6.0%) relative to CPF; C. parapsilosis (from 0.8-3.4%) and C. glabrata (from 11.0-20%) against FLC; and C. glabrata (from 3.0-12.0%) for VRC. Among the moulds, most Aspergillus spp. isolates were found to be susceptible to VRC, ITC, and POS, while AMB and POS were confirmed to be the most effective agents against zygomycetes. EUCAST and CLSI should continue their efforts to harmonize their methods of antimicrobial susceptibility testing (AST) and to define additional and shareable epidemiological cutoff values and clinical breakpoints. PMID- 21619498 TI - Genetic and biological features of catheter-associated Malassezia furfur from hospitalized adults. AB - Malassezia furfur, an etiological agent of catheter-associated fungemia, requires long-chain fatty acids for in vitro growth. We examined the applicability of rDNA sequence analysis, autoaggregation testing in liquid culture, utilization of parenteral lipid emulsions, and phospholipase activity for discrimination of catheter-associated M. furfur strains. The rDNA sequence types of catheter associated M. furfur strains were distinct from those of other isolates. All M. furfur isolates recovered from blood culture bottles and the tips of catheters from patients receiving fat emulsion therapy were type I-3. Only M. furfur isolate GIFU 01 from a blood culture bottle showed no autoaggregation in liquid culture. All strains of M. furfur examined grew well on Sabouraud's dextrose agar supplemented with Intralipid lipid emulsion as compared to individual Tweens (20, 40, 60, 80) and Cremophor EL. A high percentage of type I-3 M. furfur strains (80.0%) showed very high phospholipase activity compared to type I-1 and I-4 strains obtained from healthy skin of the same subjects or healthy control subjects (20.0% and 0.0%, respectively). The blood culture bottle isolate GIFU 01 showed very high lipolytic enzymes activity for Intralipid but no phospholipase activity. These results suggest that particular factors, such as non autoaggregation and very high lipolytic enzyme activity for parenteral lipid emulsions, play important roles in the growth and pathogenicity of Malassezia related sepsis. PMID- 21619499 TI - Statistical analysis of arthroplasty data. I. Introduction and background. PMID- 21619500 TI - Statistical analysis of arthroplasty data. II. Guidelines. PMID- 21619501 TI - Clinical outcome after undisplaced femoral neck fractures. AB - BACKGROUND AND PURPOSE: Little attention has been paid to undisplaced femoral neck fractures. By using data from the Norwegian Hip Fracture Register, we investigated the risk of reoperation and the clinical outcome after treatment of these fractures in patients over 60 years of age. METHODS: Data on 4,468 patients with undisplaced femoral neck fractures who were operated with screw osteosynthesis were compared to those from 10,289 patients with displaced femoral neck fractures treated with screw osteosynthesis (n = 3,389) or bipolar hemiarthroplasty (n = 6,900). The evaluation was based on number of reoperations and patient assessment at 4 and 12 months of follow-up. RESULTS: The 1-year implant survival was 89% after screw fixation for undisplaced fractures, 79% after screw fixation for displaced fractures, and 97% after hemiarthroplasty for displaced fractures. Patients with displaced fractures who were operated with internal fixation had a higher risk of reoperation (RR = 1.9, CI: 1.7-2.2), reported more pain, were less satisfied, and had lower quality of life than patients with undisplaced fractures treated with internal fixation (p < 0.05). Patients with displaced fractures who were operated with hemiarthroplasty had a lower risk of reoperation than patients with undisplaced fractures who were operated with internal fixation (RR = 0.32, CI: 0.27-0.38). Furthermore, they had the lowest degree of pain, were most satisfied, and reported the highest quality of life. INTERPRETATION: The differences in clinical outcome found were less than what is considered to be of clinical importance. The results support the use of screw osteosynthesis for undisplaced femoral neck fractures in elderly patients, although even better results were obtained in the hemiarthroplasty group in patients with displaced fractures. PMID- 21619502 TI - Failure of total knee arthroplasty with or without patella resurfacing. AB - BACKGROUND AND PURPOSE: Patella resurfacing during primary total knee arthroplasty (TKA) is disputed and new prosthesis designs have been introduced without documentation of their survival. We assessed the impact on prosthesis survival of patella resurfacing and of prosthesis brand, based on data from the Norwegian Arthroplasty Register. PATIENTS AND METHODS: 5 prosthesis brands in common use with and without patella resurfacing from 1994 through 2009 were included n = 11,887. The median follow-up times were 9 years for patella resurfaced implants and 7 years for implants without patella resurfacing. For comparison of prosthesis brands, also brands in common use with only one of the two treatment options were included in the study population (n = 25,590). Cox regression analyses were performed with different reasons for revision as endpoints with adjustment for potential confounders. RESULTS: We observed a reduced overall risk of revision for patella resurfaced (PR) TKAs, but the statistical significance was borderline (RR = 0.84, p = 0.05). At 15 years, 92% of PR and 91% of patella non resurfaced (NR) prostheses were still unrevised. However, PR implants had a lower risk of revision due to pain alone (RR = 0.1, p < 0.001), but a higher risk of revision due to loosening of the tibial component (RR = 1.4, p = 0.03) and due to a defective polyethylene insert (RR = 3.2, p < 0.001). At 10 years, the survival for the reference NR brand AGC Universal was 93%. The NR brands Genesis I, Duracon, and Tricon (RR = 1.4-1.7) performed statistically significantly worse than NR AGC Universal, while the NR prostheses e.motion, Profix, and AGC Anatomic (RR = 0.1-0.7), and the PR prostheses NexGen and AGC Universal (RR = 0.4-0.5) performed statistically significantly better. LCS, NexGen, LCS Complete (all NR), and Tricon, Genesis I, LCS, and Kinemax (all PR) showed no differences in this respect from the reference brand. A lower risk of revision (crude) was found for TKAs performed after 2000 as compared to those performed earlier (RR = 0.8, p = 0.001). INTERPRETATION: Although revision risk was similar for PR and NR TKAs, we found important differences in reasons for revision. Our results also indicate that survivorship of TKAs has improved. PMID- 21619503 TI - No evidence of a clinically important effect of adding local infusion analgesia administrated through a catheter in pain treatment after total hip arthroplasty. AB - BACKGROUND AND PURPOSE: Postoperative analgesia after primary total hip arthroplasty (THA) using opioids is associated with troublesome side effects such as nausea and dizziness, and epidural analgesic means delayed mobilization. Thus, local infiltration analgesia (LIA) during surgery prolonged with local infusion analgesia (LINFA) into the soft tissue in the hip region through a catheter in the first postoperative days has gained major interest in THA fast-track settings within a short period of time. LIA at the time of surgery is a validated treatment. We investigated the additional effect of giving postoperative LINFA after THA in patients already having LIA during surgery. PATIENTS AND METHODS: 60 consecutive patients undergoing non-cemented THA were randomized into two groups in a double-blind and controlled study. During surgery, all patients received standardized pain treatment with LIA. Postoperatively, they were treated either with a solution of Ropivacain, Ketorolac, and Adrenaline (LINFA group) or placebo (placebo group) administered through a catheter to the hip 10 and 22 h after surgery. Pain score, opioid consumption, and length of stay (LOS) were evaluated. RESULTS: After adjustment for multiple testing, there was no statistically significant postoperative difference between the LINFA group and the placebo group regarding pain and tiredness. We found some evidence of a short-term effect on nausea and vomiting. Opioid consumption and length of stay were similar in the two groups. INTERPRETATION: We found some evidence of a short-term effect of LINFA on nausea and vomiting, but no evidence of an effect on postoperative pain and tiredness. Thus, LINFA cannot be recommended as a standard pain treatment in patients with THA. PMID- 21619504 TI - Changing from analog to digital images: does it affect the accuracy of alignment measurements of the lower extremity? AB - BACKGROUND AND PURPOSE: Medical imaging has changed from analog films to digital media. We examined and compared the accuracy of orthopedic measurements using different media. METHODS: Before knee arthroplasty, full-length standing radiographs of 52 legs were obtained. The mechanical axis (MA), tibio-femoral angle (TFA), and femur angle (FA) were measured and analyzed twice, by 2 radiologists, using (1) true-size films, (2) short films, (3) a digital high resolution workstation, and (4) a web-based personal computer. The agreement between the 4 media was evaluated using the Bland-Altman method (limits of agreement) using the true-size films as a reference standard. RESULTS: The mean differences in measurements between the traditional true-size films and the 3 other methods were small: for MA -0.20 to 0.07 degrees, and for TFA -0.02 to 0.18 degrees. Also, the limits of agreement between the traditional true-size films and the three other methods were small. INTERPRETATION: The agreement of the alignment measurements across the 4 different media was good. Orthopedic angles can be measured as accurately from analog films as from digital screens, regardless of film or monitor size. PMID- 21619505 TI - Totally laparoscopic gastrectomy for gastric cancer: meta-analysis of short-term outcomes. AB - INTRODUCTION: We present a review of the literature, together with a meta analysis of short-term outcomes of totally laparoscopic gastrectomy (TLG) compared with open gastrectomy (OG). MATERIAL & METHODS: We carried out a search in the Pubmed and Cochrane databases from September 2003 to May 2009. Controlled studies on early outcomes were included, both prospective and retrospective, randomized and non-randomized. RESULTS: We found nine eligible studies, one of which was a randomized controlled trial (RCT), while eight were series of patients (three consecutive). The study group consisted of 1,492 patients, 828 of whom had been treated with TLG and 664 treated with OG. TLG for gastric cancer shows a 32.5% (p < 0.001) longer operative time than OG, whereas TLG demonstrated a 44% (p < 0.001) reduction in blood loss, a 34% (p < 0.001) reduction time to first flatus and a 33.7% reduced (p < 0.001) hospital stay. No notable differences were registered regarding morbidity and mortality rates, and no significant difference was observed between the two groups regarding the extent of the lymphadenectomy. CONCLUSIONS: Despite a longer operative time for TLG, with a gastrointestinal recovery rate faster than the OG one for gastric cancer results, no notable differences were recorded between the two techniques for the morbidity and mortality rates and in the spread of the lymphadenectomy. PMID- 21619506 TI - Degradable poly(amido amine)s as gene delivery carriers. AB - INTRODUCTION: In recent years, there has been a great deal of interest in the development of vectors which are being developed based on the capacity of polymers to mediate appropriate interactions with the cellular environment, or to interface with specific cellular processes. Several such vectors have been synthesized, resulting in biomacromolecules with low cytotoxicity and higher gene delivery ability. AREAS COVERED: This review briefly describes the recent success of poly(amido amine)s (PAAs) as non-viral vectors, and highlights their promising future in the development of nucleic acid-based therapy. It also provides an overview on the synthesis, characterization and application of PAAs as gene carriers, which will be useful for various biological motifs. This review helps the readers to better understand the emergence of non-viral vectors for gene therapy, especially PAAs, their properties, their advantages and disadvantages and the gene therapy based on them. EXPERT OPINION: The future of gene-based therapy needs to identify approaches to develop new carriers, depending on the properties of the biological membranes they face, and their physicochemical properties, in order to successfully deliver the genes to the target sites. With the emergence of a variety of non-viral vectors, such as biodegradable polymers, it may not take long before non-viral vectors are observed that are not just safe and tissue-specific, but even more efficient than viral vectors. PMID- 21619507 TI - Therapeutic management and clinical outcome of autoimmune pancreatitis. AB - BACKGROUND: Autoimmune pancreatitis, in comparison to other benign chronic pancreatic diseases, is characterized by the possibility of curing the illness with immunosuppressant drugs. The open question is whether to differentiate autoimmune pancreatitis as a primary or secondary disease based on the presence or absence of other autoimmune diseases or whether to consider autoimmune pancreatitis a clinical and pathological systemic entity, called IgG4-related sclerosing disease, since this aspect is also very important from a therapeutic point of view. METHODS: In this paper, we reviewed the conventional therapeutic approach used to treat autoimmune pancreatitis patients and the clinical outcome related to each treatment modality. We also reviewed some aspects which are important for the correct management of autoimmune pancreatitis, such as the surgical approach, the outcome of surgically treated autoimmune pancreatitis patients, whether medical treatment is always necessary, and, finally, when medical treatment should be initiated. CONCLUSIONS: Steroids are useful in alleviating the symptoms of the acute presentation of autoimmune pancreatitis, but some questions remain open such as the dosage of steroids in the acute phase and the duration of steroid therapy; finally, it should be assessed if other immunosuppressive non-steroidal drugs may become the first-line therapy in patients with AIP without jaundice and without atrophic pancreas. PMID- 21619508 TI - Macromolecular bases of antischistosomal therapy. AB - Schistosomiasis is a widespread tropical parasitic disease, currently treated with Praziquantel, whose precise molecular target is actually unknown. Several other drugs are known to kill the schistosomes in vivo and in vitro, but these are seldom employed because of toxicity, high cost, complex administration or other reasons. The improvement of known drugs or the development of entirely new ones is a desirable goal, in view of the fact that strains of Schistosoma mansoni with reduced sensitivity to Praziquantel have appeared. In this review, we tried to collect the information available on known or putative macromolecular targets of schistosomicidal drugs; thus we focused on the biochemistry of the parasite, rather than the clinical properties of the drugs. The rationale of this approach is that drug design may become realistic if the mechanism of action of each known drug were known at atomic detail, ideally as the 3D structure of each drug in complex with its target. Important macromolecular targets of known drugs reviewed below are: Thioredoxin Glutathione Reductase; Cyclophilin; Acetyl Cholinesterase; Proteases and Purine Nucleoside Phosphorylase. Moreover, a few enzymes of the parasite are known, or thought, to be "druggable", and therefore interesting, even though no specific drugs are available as yet: examples of such enzymes are Glutathione Peroxidase and Peroxiredoxins. PMID- 21619509 TI - Isoprenoid precursor biosynthesis offers potential targets for drug discovery against diseases caused by apicomplexan parasites. AB - Two, simple, C5 compounds, dimethylally diphosphate and isopentenyl diphosphate, are the universal precursors of isoprenoids, a large family of natural products involved in numerous important biological processes. Two distinct biosynthetic pathways have evolved to supply these precursors. Humans use the mevalonate route whilst many species of bacteria including important pathogens, plant chloroplasts and apicomplexan parasites exploit the non-mevalonate pathway. The absence from humans, combined with genetic and chemical validation suggests that the non mevalonate pathway holds the potential to support new drug discovery programmes targeting Gram-negative bacteria and the apicomplexan parasites responsible for causing serious human diseases, and also infections of veterinary importance. The non-mevalonate pathway relies on eight enzyme-catalyzed stages exploiting a range of cofactors and metal ions. A wealth of structural and mechanistic data, mainly derived from studies of bacterial enzymes, now exists for most components of the pathway and these will be described. Particular attention will be paid to how these data inform on the apicomplexan orthologues concentrating on the enzymes from Plasmodium spp. these cause malaria, one the most important parasitic diseases in the world today. PMID- 21619510 TI - Trypanocidal activity of nitroaromatic prodrugs: current treatments and future perspectives. AB - Chagas disease and African sleeping sickness are trypanosomal infections that represent important public health problems in Latin America and Africa, respectively. The restriction of these diseases to the poorer parts of the world has meant that they have been largely neglected and limited progress has been made in their treatment. The nitroheterocyclic prodrugs nifurtimox and benznidazole, in use against Chagas disease for >40 years, remain the only agents available for this infection. In the case of African sleeping sickness, nifurtimox has recently been added to the arsenal of medicines, with the nitroheterocycle fexinidazole currently under evaluation. For a long time, the cytotoxic mechanism of these drugs was poorly understood: nifurtimox was thought to act via production of superoxide anions and nitro radicals, while the mode of benznidazole action was more obscure. The trypanocidal activity of nitroheterocyclic drugs is now known to depend on a parasite type I nitroreductase (NTR). This enzyme is absent from mammalian cells, a difference that forms the basis for the drug selectivity. The role of this enzyme in drug activation has been genetically and biochemically validated. It catalyses the 2 electron reduction of nitroheterocyclic compounds within the parasite, producing toxic metabolites without significant generation of superoxide. Recognition that this enzyme is responsible for activation of nitroheterocyclic prodrugs has allowed screening for compounds that preferentially target the parasite. This approach has led to the identification of two new classes of anti-trypanosomal agents, nitrobenzylphosphoramide mustards and aziridinyl nitrobenzamides, and promises to yield new, safer, more effective drugs. PMID- 21619511 TI - Purine and pyrimidine pathways as targets in Plasmodium falciparum. AB - Malaria is a leading cause of morbidity and mortality in the tropics. Chemotherapeutic and vector control strategies have been applied for more than a century but have not been efficient in disease eradication. Increased resistance of malaria parasites to drug treatment and of mosquito vectors to insecticides requires the development of novel chemotherapeutic agents. Malaria parasites exhibit rapid nucleic acid synthesis during their intraerythrocytic growth phase. Plasmodium purine and pyrimidine metabolic pathways are distinct from those of their human hosts. Thus, targeting purine and pyrimidine metabolic pathways provides a promising route for novel drug development. Recent developments in enzymatic transition state analysis have provided an improved route to inhibitor design targeted to specific enzymes, including those of purine and pyrimidine metabolism. Modern transition state analogue drug discovery has resulted in transition state analogues capable of binding to target enzymes with unprecedented affinity and specificity. These agents can provide specific blocks in essential pathways. The combination of tight binding with the high specificity of these logically designed inhibitors, results in low toxicity and minor side effects. These features reduce two of the major problems with the current antimalarials. Transition state analogue design is being applied to generate new lead compounds to treat malaria by targeting purine and pyrimidine pathways. PMID- 21619512 TI - Apical membrane antigen 1 as an anti-malarial drug target. AB - The invasion of host cells by malaria parasites represents an attractive target for therapeutic intervention. The role played by apical membrane antigen 1 (AMA1) in this process has been elucidated recently with the demonstration that AMA1 forms a complex with parasite rhoptry neck (RON) proteins as part of the moving junction that develops between the host cell and the invading parasite. Structural studies of AMA1 alone and in complexes with antibodies that inhibit host cell invasion have identified a conserved hydrophobic cleft that is essential to the assembly of the AMA1/RON complex. AMA1 is already established as an important candidate for inclusion in a malaria vaccine. Here we review both the structural details and functional significance of interactions at the hydrophobic cleft of AMA1, and argue that this feature of the protein represents an excellent target for the development of drugs that would block host cell invasion by malarial parasites. PMID- 21619513 TI - Sterol 14alpha-demethylase (CYP51) as a therapeutic target for human trypanosomiasis and leishmaniasis. AB - Pathogenic protozoa threaten lives of several hundred million people throughout the world and are responsible for large numbers of deaths globally. The parasites are transmitted to humans by insect vectors, more than a hundred of infected mammalian species forming reservoir. With human migrations, HIV-coinfections, and blood bank contamination the diseases are now spreading beyond the endemic tropical countries, being found in all parts of the world including the USA, Canada and Europe. In spite of the widely appreciated magnitude of this health problem, current treatment for sleeping sickness (Trypanosoma brucei), Chagas disease (Trypanosoma cruzi) and leishmaniasis (Leishmania spp.) remains unsatisfactory. The drugs are decades old, their efficacy and safety profiles are unacceptable. This review describes sterol 14alpha-demethylase, an essential enzyme in sterol biosynthesis in eukaryotes and clinical target for antifungal azoles, as a promising target for antiprotozoan chemotherapy. While several antifungal azoles have been proven active against Trypanosomatidae and are under consideration as antiprotozoan agents, crystal structures of sterol 14alpha demethylases from three protozoan pathogens, Trypanosoma brucei, Trypanosoma cruzi and Leishmania infantum provide the basis for the development of new, highly potent and pathogen-specific drugs with rationally optimized pharmacological properties. PMID- 21619514 TI - Drug target identification in intracellular and extracellular protozoan parasites. AB - The increasing demand for novel anti-parasitic drugs due to resistance formation to well-established chemotherapeutically important compounds has increased the demands for a better understanding of the mechanism(s) of action of existing drugs and of drugs in development. While different approaches have been developed to identify the targets and thus mode of action of anti-parasitic compounds, it has become clear that many drugs act not only on one, but possibly several parasite molecules or even pathways. Ideally, these targets are not present in any cells of the host. In the case of apicomplexan parasites, the unique apicoplast, provides a suitable target for compounds binding to DNA or ribosomal RNA of prokaryotic origin. In the case of intracellular pathogens, a given drug might not only affect the pathogen by directly acting on parasite-associated targets, but also indirectly, by altering the host cell physiology. This in turn could affect the parasite development and lead to parasite death. In this review, we provide an overview of strategies for target identification, and present examples of selected drug targets, ranging from proteins to nucleic acids to intermediary metabolism. PMID- 21619515 TI - 6-oxopurine phosphoribosyltransferase: a target for the development of antimalarial drugs. AB - Malaria remains the most serious parasitic diseases affecting humans in the world today, resulting in 1-2 million fatalities each year. Plasmodium falciparum (Pf) and Plasmodium vivax (Pv) are the predominant causative agents. Both are responsible for widespread mortality and morbidity and are a serious socio economic burden, especially for countries in the developing world. One of the most important defences against malaria has been the use of chemotherapeutic drugs (e.g. chloroquine, artemisinins, pyrimethamine) but these have mainly been found by serendipity. Their mechanisms was not understood at the time of their discovery and, even today, are still not unequivocal. For many of these compounds, the parasite is now resistant and, hence, there is an urgent need to develop new therapeutic drugs directed to validated targets. One metabolic pathway crucial for the survival and replication and survival of the parasite is the synthesis of the purine nucleoside monophosphates essential for the production of DNA/RNA molecules. A key enzyme in this pathway is the 6-oxopurine phosphoribosyltransferase (PRTase). The focus of this review is on the identification and characterization of inhibitors of the enzymes from both Pf and Pv as antimalarial drug leads. The acyclic nucleoside phosphonates (ANPs) appear to be excellent candidates because they are good inhibitors of the two Plasmodium enzymes, can be selective compared to the human enzyme, can arrest parasitemia in cell based assays, have low cytotoxicity to the human host cell and, because of their stable carbon-phosphorous bond, are stable within the cell. PMID- 21619516 TI - Drug targets for the treatment of protozoan parasitic diseases. PMID- 21619517 TI - Association and causal relationship of midlife obesity and related metabolic disorders with old age cognition. AB - OBJECTIVE: Epidemiological studies suggest a relationship between midlife metabolism and old age cognition. We examined the effect of midlife BMI and related metabolic conditions on old age cognitive performance and whether there was evidence from direct causal pathways behind these associations in a large sample of Finnish twins. DESIGN: Midlife variables of 2606 twin individuals were based on postal questionnaires and registry records. Old age cognitive status was measured by using a validated telephone interview. RESULTS: Midlife BMI, cardiovascular disease, hypertension and diabetes were each associated with old age cognition when adjusted for sex, education, birth year and age at the interview. Similarly, overweight increased the risk for categories of mild impairment of cognitive function and likely dementia. Cardiovascular disease diminished the mean cognitive score also among discordant twin pairs (beta estimate=1.10, p-value= 0.012). Weight gain more than 1.7 kg/m(2) and loss more than 2 kg/m(2) within an average of 5.6 years were associated with lower cognitive performance independently of BMI. An additive genetic correlation explained the association between BMI and old age cognition (r(A)=-0.12, 95% CI 0.21; -0.03), but adjustment for education led to loss of significance (r(A)= 0.06, 95% CI -0.16; 0.03). CONCLUSIONS: Midlife metabolic diseases, especially diabetes, are independently associated with impaired cognition in old age. Even a more subtle weight change than suggested previously was associated with lower old age cognition. There was evidence from direct causal pathway between cardiovascular disease and old age cognition, while the correlation between midlife BMI and old age cognition was explained mostly by genetic factors. PMID- 21619518 TI - Herb-drug interactions involving drug metabolizing enzymes and transporters. AB - Herbal medicines and their active ingredients are widely used worldwide, and they have become an important part of clinical medicine. The combined use of herbs and drugs has increased the possibility of pharmacokinetic and pharmacodynamic interactions. Clinical studies have demonstrated that the combined use of herbs and drugs can enhance or attenuate the drug efficacy and toxicity. The herb-drug combinations may reduce a drug efficacy and lead to treatment failure when long term administration. Case reports detailing serious clinical adverse reactions have promoted studies on the interactions between herbs and drugs. This review highlights recent knowledge to discuss herb-drug interactions involving metabolizing enzymes and drug transporters. Drug transporters are widely present in body and play an important role in the absorption, distribution, excretion and metabolism, efficacy, and toxicity of drugs. Investigation of transporters has developed rapidly since 1990s, the effects of many transporters on the pharmacokinetics of drugs and herb-drug interactions have been reported. Some concepts on drug transporters issued experimentally and clinically drug-drug and herb-drug interactions have applied in many studies. Methodology studies are very important for understanding the mechanism, considerations and evaluation of experiments and clinical studies on drug metabolizing enzymes and transporters in drug-drug interactions. PMID- 21619519 TI - Metabolism of ginseng and its interactions with drugs. AB - Ginseng is an herbal medicine used worldwide. It is reported to have a wide range of pharmacological activities because of a diversified group of steroidal saponins called ginsenosides. Compared to extensive pharmacological studies of ginseng, the pharmacokinetics, especially the metabolism of this herb, has received less attention. In this article we review the known pharmacokinetic data on ginseng. Understanding ginseng's pharmacokinetics may reduce the potential for interactions in patients who use both ginseng and prescription medications. In addition, bioavailability after taking ginseng orally is low, and the metabolites of ginsenosides produced by gut microbiota may be biologically active. One ginseng metabolite, Compound K, and its potential for cancer chemoprevention is also discussed. An active ginseng metabolite may differ in distribution and clearance from its parent compound, and the parent compound and its metabolite may be bioactive by similar or different mechanisms. Thus, further investigation of ginseng metabolites is needed for predicting the therapeutic outcome with ginseng. PMID- 21619520 TI - Hepatotoxicity and tumorigenicity induced by metabolic activation of pyrrolizidine alkaloids in herbs. AB - In the recent decades, the use of herbal products has been rapidly growing in the Western countries. While their use in many cases causes adverse effects, to date, safety issues of herbal products have not been adequately addressed. It is rarely determined whether the non-purported bioactive constituents in the herbs and the metabolites of the bioactive components can lead to adverse effects. In this review, we discuss, using pyrrolizidine alkaloids (PAs) as an example, the hepatotoxicity and tumorigenicity induced by metabolic activation of herbal components and by herb-herb and herb-drug interactions with other herbal ingredients and synthetic drugs. PAs are constitutively produced by plants as the secondary metabolites. There are more than 600 PAs and PA N-oxides identified in over 6000 plants, and more than half of them exhibit hepatotoxicity. Toxic PA containing plants grow in many geographical regions worldwide, rendering it highly possible that PA-containing plants are the most common poisonous plants affecting livestock and humans. PAs require metabolic activation mediated by cytochrome P450 enzymes to generate reactive pyrrolic metabolites that react with cellular proteins and DNA leading to hepatotoxicity and genotoxicity. PAs can also modulate both phase I and phase II metabolizing enzymes, which may alter the metabolic fate of endogenous and exogenous chemicals. Alteration and/or competition of the metabolizing enzymes by PAs upon the co-administered herbal medicines or drugs can potentially result in serious clinical and toxicological consequences through decreased pharmacological activities or increased toxic effects. PMID- 21619521 TI - Strategies for integral metabolism profile of multiple compounds in herbal medicines: pharmacokinetics, metabolites characterization and metabolic interactions. AB - Herbal medicines (HMs) are gaining more and more attention all over the world, because of their significant curative effect in treating multi-factorial diseases. Recently, the in vivo and in vitro metabolism study of HMs has become an important issue because these data can help us to better understand the efficacies and toxicities of HMs. However, the integral metabolism profile of HMs is confronted with many challenges: 1) HM is a multi-component system; 2) most components are unknown (nontarget); 3) trace of components in HM. Given the challenges described above, the demand for more powerful bioanalytical tools and strategies that are adequate for integral metabolism profile of HMs' multi components has increased. In the past few years, newer methods, or adaptations to methods, have been published, and this review will attempt to discuss new improvements in strategies and methodologies for HMs' multi-component ADME evaluation. In particular, improvements have been reported for experimental approaches to pharmacokinetics study of HMs, as well as strategies applied to metabolites characterization of HMs' multi-components, and the metabolic interactions between ingredients in HMs, including advance and proposed strategy: "chemical fishing" based strategy for metabolic interactions of HMs. PMID- 21619522 TI - Prediction of drug-drug interactions arising from mechanism-based inactivation: key input parameters and impact on risk assessment. AB - As the prevalence of polypharmacy increases with our aging population, the propensity for adverse drug-drug interactions arising from the altered metabolism of co-administered medicines remains an important consideration for drug development. Mechanism-based inactivation (MBI) of the cytochrome P450 enzyme system is responsible for many clinically relevant drug-drug interactions (DDIs) due to the irreversible and long-lasting effects of the enzyme inactivation. Unlike competitive inhibition, MBI persists after the inactivator has been cleared from the system, since de novo enzyme synthesis is required to restore metabolic activity. Recognizing the potential severity of DDIs arising from MBI, there is increasing need for predictive methodologies that can enable prospective risk assessment for the likelihood of a clinical DDI. Steady-state models, which simplify the system to a single inactivator concentration and assume static, equilibrium conditions, are important tools for assessing the potential for DDIs. More sophisticated, physiologically-based models offer advantages over the static models by taking into account changing inactivator concentration over time, in addition to incorporating population variability into the prediction. Despite the differences between the static and dynamic approaches, a key consideration for both is the sensitivity of the models to the input parameters. These inputs include inactivator-specific kinetic parameters describing MBI in terms of potency (K(I)) and inactivation rate (k(inact)), the unbound inactivator concentration (I(u)), and the enzyme degradation rate, (k(deg)). This commentary investigates the impact of the selection of input parameters, and the uncertainty in their assessment, on the prediction for DDIs arising from MBI and the relevance to risk-assessment. PMID- 21619523 TI - Translational research insights into pharmacokinetic herb-drug interactions. AB - Translational research is an emerging discipline that aims to fill the gap through the whole pipeline from bench to bedside. This review highlights the importance and urgency of incorporating translational research into the study of pharmacokinetic herb drug interactions (PHDI), based on an intensive discussion on the controversial and inconsistent reports from preclinical to clinical, in vitro to in vivo, and across different studies concerning PHDI. Current controversial and dispersed reports confer poor translational capacity of experimental research data to guiding the clinical practices. We propose that the herbal complexities in their chemical compositions, biphasic and tissue-specific effects of enzymes, and the present incomplete understanding of the disposition properties of herbal medicines themselves; and the enzymatic complexities in the species differences, individual genotype and phenotype, differential regulation during healthy and pathological conditions, substrate dependent modulations, and their interplay with transporters, collectively constitute the major translational blocks in PHDI from experimental research to daily clinical practice. In clinical considerations, this review indicates that PHDI are far from clear based on the isolated preclinical findings, and that the intestine is a much more susceptible site than liver when subjected to herbal regulations, and that enzymatic induction could be more predominant than inhibition upon chronic ingestion of herbal medicines. Hopefully, this review would be helpful for better understanding the nature of hurdles in PHDI research, and for igniting the future translational research initiatives. PMID- 21619524 TI - Drug design by generalized-ensemble simulations. AB - In pharmaceutical design based on molecular simulations, one faces a great difficulty. Conventional simulations in the canonical ensemble are of little use, because they tend to get trapped in some of the local-minimum-energy states that exist in a huge number. A simulation in generalized ensemble performs a random walk in potential energy space and can overcome this difficulty. In this article we review a few of powerful generalized-ensemble algorithms, namely, replica exchange method, multidimensional replica-exchange method, and replica-exchange umbrella sampling. The effectiveness of the methods is tested with ligand and short peptide systems. PMID- 21619525 TI - Membrane targeted chemotherapy with hybrid liposomes for tumor cells leading to apoptosis. AB - We have produced hybrid liposomes (HL) which can be prepared by sonication of a mixture of vesicular and micellar molecules in a buffer solution. The physical properties of HL such as size, shape, and membrane fluidity can be controlled by changing the constituents and compositional ratio. We have employed HL for chemotherapy and interesting results are as follows; (A) The uniform and stable structure of HL composed of L-alpha-dimyristoylphosphatidylcholine (DMPC) and polyoxyethylenedodecyl ether (C12(EO)(n)) with a diameter of 80 nm was revealed. (B) The remarkable inhibitory effects of HL on the growth of various tumor cells were attained in vitro. (C) Induction of apoptosis by HL was obtained and the pathway of apoptosis induced by HL was clarified. (D) A good correlation between the membrane fluidity of HL and inhibitory effects of HL for tumor cells was obtained. (E) Significantly chemotherapeutic effects were obtained using mice model of carcinoma after the treatment with HL without any side effects in vivo. (F) In clinical applications, prolonged survival and remarkable reduction of neoplasm were attained in patients with lymphoma after the treatment with HL without any side effects after the approval of the bioethics committee. PMID- 21619527 TI - A novel ligand-mapping method based on molecular liquid theory. AB - The recent development of a novel ligand-mapping method is reviewed. The method is based on a statistical-mechanical molecular theory of solvation, known as the three-dimensional reference interaction site model (3D-RISM). In the 3D-RISM based ligand mapping (3D-RISM-LM) method, using the all-atom model for a target protein immersed in a ligand-water mixture solvent, the 3D-spatial distributions of the ligand atomic sites around the protein are first obtained, and then the most probable binding modes of the ligand molecule are constructed from the distributions. Unlike conventional docking simulations, 3D-RISM-LM can incorporate the effect of water from the atomic to thermodynamic level into the binding affinity through statistical mechanics. It has been demonstrated that 3D RISM-LM can sensitively detect even weak binding modes of small molecules over the entire surface of protein. Therefore, this approach is expected to be particularly useful in fragment-based drug design. PMID- 21619528 TI - Structural and molecular basis of carbohydrate-protein interaction systems as potential therapeutic targets. AB - The sugar chains covalently modifying proteins and lipids are recognized by a variety of proteins, thereby mediating a broad range of physiological and pathological events on cell surfaces as well as in cells. Hence, these carbohydrate-protein interaction systems could be potential therapeutic targets for various diseases, including viral infections, autoimmune diseases and neurodegenerative disorders. Cumulative crystallographic data of lectins complexed with their cognate carbohydrate ligands have elucidated the sugar recognition modes of these proteins, offering a structural basis for the design of drugs targeting carbohydrate-lectin interaction systems. In particular, structural and functional studies of animal L-type lectins, which possess a carbohydrate recognition domain with a structural resemblance to that of leguminous lectins such as concanavalin A, have demonstrated the molecular mechanisms underlying their distinct roles in sorting and trafficking of glycoproteins in cells, exemplifying the structure-based engineering that manipulates the sugar-binding properties of lectins. Furthermore, structural basis has been provided for the functional interplay between the L-type lectin ERGIC-53 and the EF-hand Ca2+-binding protein MCFD2 in the intracellular transport of the coagulation factors V and VIII. This article also deals with pathological carbohydrate-protein interactions involving ganglioside clusters on cell surfaces, particularly focusing on the interaction between amyloid beta (Abeta) and GM1 ganglioside. This interaction triggers conformational transition and consequent aggregation of Abeta, and therefore, is considered to be a key step in Alzheimer's disease. The recently reported structural information of the Abeta-GM1 interaction is presented, underscoring the significance of assemblages of glycoconjugates as therapeutic targets. PMID- 21619526 TI - Molecular recognition in the case of flexible targets. AB - A protein's flexibility is well recognized to underlie its capacity to engage in critical functions, such as signal transduction, biomolecular transport and biochemical reactivity. Molecular recognition is also tightly linked to the dynamics of the binding partners, yet protein flexibility has largely been ignored by the growing field of structure-based drug design (SBDD). In combination with experimentally determined structures, a number of computational methods have been proposed to model protein movements, which may be important for small molecule binding. Such techniques have the ability to expose new binding site conformations, which may in turn recognize and lead to the discovery of more potent and selective drugs through molecular docking. In this article, we discuss various methods and focus on the Relaxed Complex Scheme (RCS), which uses Molecular Dynamics (MD) simulations to model full protein flexibility and enhance virtual screening programmes. We review practical applications of the RCS and use a recent study of the HIV-1 reverse transcriptase to illustrate the various phases of the scheme. We also discuss some encouraging developments, aimed at addressing current weaknesses of the RCS. PMID- 21619529 TI - Computational studies of influenza A virus at three important targets: hemagglutinin, neuraminidase and M2 protein. AB - While the seasonal influenza viruses spreading around the world cause the annual epidemics, the recent outbreaks of influenza A virus subtype H5N1 and pandemic H1N1 have raised a global human health concerns. In this review, the applicability of computational techniques focused on three important targets in the viral life cycle: hemagglutinin, neuraminidase and M2 proton channel are summarized. Protein mechanism of action, substrate binding specificity and drug resistance, ligand-target interactions of substrate/inhibitor binding to these three proteins either wild-type or mutant strains are discussed and compared. Advances on the novel anti-influenza agents designed specifically to combat the avian H5N1 and pandemic H1N1 viruses are introduced. A better understanding of molecular inhibition and source of drug resistance as well as a set of newly designed compounds is greatly useful as a rotational guide for synthetic and medicinal chemists to develop a new generation of anti-influenza drugs. PMID- 21619530 TI - Molecular recognition explored by a statistical-mechanics theory of liquids. AB - "Molecular recognition" is one of the most important molecular processes for living systems in order to maintain their life, since most of the biological functions are initiated with the process. Understanding of the process is also important for designing a new drug. Firstly, it is important to find a target of a drug, which is in many cases a function of protein or DNA to be inhibited. Secondly, binding a drug molecule to the active site of a biomolecule itself is a "molecular recognition process". In the present article, we review our recent studies on the molecular recognition process, carried out by means of the 3D-RISM theory, a statistical mechanics theory of liquids. Studies on the conduction mechanisms in two types of molecular channels, aquaporin and the M2 channels, are reviewed. PMID- 21619531 TI - Structural basis of agonist selectivity for different nAChR subtypes: insights from crystal structures, mutation experiments and molecular simulations. AB - Nicotinic acetylcholine receptors (nAChRs) are members of ligand gated ion channels (LGICs) which transduce chemical signal into electrical signal in neuron and neuromuscular junction. They are pentamerics which contain an extra-cellular domain (also known as ligand binding domain or LBD), a trans-membrane domain and a cytoplasmic domain (intra-cellular domain). Agonist binding to the extra cellular domain invokes positive ion flux as well as action potential in neurons, muscle cells and endocrine cells whereas antagonist binding inhibits ion flux. There are various endogenous or exogenous compounds which behave as agonists or antagonists targeting nAChRs. During the last decades, the whole structure of muscle type nAChR as well as the crystal structures of acetylcholine-binding proteins (AChBPs) which are homologues of the nAChRs extra-cellular domain has been obtained. These structures, together with other studies including mutation experiments and molecular simulations, provide insights into both of the nAChR architecture and its agonist binding cavity. Our review gives detailed accounts of the recent progresses in order to gain insights into agonist selectivity for different nAChR subtypes. PMID- 21619532 TI - In silico screening of bioactive and biomimetic solutes using Integral Equation Theory. AB - The Integral Equation Theory (IET) of Molecular Liquids is a theoretical framework for modelling solution phase behaviour that has recently found new applications in computational drug design. IET allows calculation of solvation thermodynamic parameters at significantly lower computational expense than explicit solvent simulations, but also provides information about the microscopic solvent structure that is not accessible by implicit continuum models. In this review we focus on recent advances in two fields of research using these methods: (i) calculation of the hydration free energies of bioactive molecules; (ii) modelling the aggregation of biomimetic molecules. In addition, we discuss sources of experimental solvation data for druglike molecules. PMID- 21619533 TI - Transforming growth factor beta in pancreatic cancer. AB - Pancreatic cancer has high incidence and mortality rates, and effective treatment remains a clinical challenge. As deregulation of the cytokine transforming growth factor beta (TGF-beta) contributes to the progression of pancreatic carcinoma, the TGF-beta pathway has been targeted using various strategies, including small molecule inhibitors of TGF-betaRI, TGF-beta-specific neutralizing antibodies and antisense compounds. As increased TGF-beta2 levels in serum or tumor tissue of patients with pancreatic cancer correlated with poor prognosis, inhibition of TGF beta2 synthesis via the antisense oligonucleotide trabedersen (AP 12009) is a promising approach. PMID- 21619534 TI - TGF-beta receptor signaling pathways in angiogenesis; emerging targets for anti angiogenesis therapy. AB - Angiogenesis, the formation of new blood vessels is essential for diverse physiological processes such as development, but also for pathological conditions like tumor growth. Most studied in this context are tyrosine kinase signaling pathways such as those involving vascular endothelial growth factor (VEGF). There is however accumulating evidence that more pathways are as essential for angiogenesis. Knockout studies of factors in transforming growth factor beta (TGF beta) signaling have for example showed that also this pathway is indispensable for angiogenesis. This review highlights our understanding of TGF-beta signaling in vascular development and angiogenesis. In particular, we focus on recent insights into the role of the TGF-beta type I receptor ALK1 and co-receptor endoglin in tumor angiogenesis, which provide opportunities for the development of new anti-angiogenesis therapies for treatment of cancer patients. PMID- 21619535 TI - Antibody targeting of TGF-beta in cancer patients. AB - The role of TGF-beta in tumor development and progression is complex. Genetic mutations that disrupt the antiproliferative signaling effects of TGF-beta play a key role in the process of malignant transformation for many types of tumors. Paradoxically, this loss of sensitivity to TGF-beta's inhibitory actions often leads to TGF-beta overexpression by the tumor cells or by normal cells that are recruited to the tumor microenvironment. Elevated concentrations of TGF-beta in the tumor microenvironment have been shown to facilitate tumor growth and metastasis. Numerous published studies have provided evidence that inhibition of TGF-beta using antibodies, soluble receptors and small molecule inhibitors of TGF beta signal transduction can have beneficial effects in murine models of cancer. Given the pleiotropic nature of TGF-beta and its homeostatic role in numerous biological processes, serious concerns have been expressed regarding the safety of administering TGF-beta antagonists to human patients. Interestingly, the results of numerous animal toxicology studies of TGF-beta antibodies in normal rodents and primates have shown that administration of neutralizing anti-TGF-beta antibodies is well tolerated and any adverse effects were reversible or self limiting. Likewise, administration of a human anti-TGF-beta antibody (fresolimumab) in three separate human phase 1 clinical trials has also been shown to be well tolerated. PMID- 21619536 TI - The antisense oligonucleotide trabedersen (AP 12009) for the targeted inhibition of TGF-beta2. AB - Despite remarkable advances in cancer research, patients with malignant tumors such as high-grade glioma or advanced pancreatic carcinoma still face a poor prognosis. Because of the severe morbidity and mortality of such malignant tumor types, the identification of suitable molecular drug targets for causal treatment approaches is an important area of current research. Transforming growth factor beta 2 (TGF-beta2) is an attractive target because it regulates key mechanisms of carcinogenesis, in particular immunosuppression and metastasis, and is frequently overexpressed in malignant tumors. Here we describe the development of the antisense phosphorothioate oligodeoxynucleotide trabedersen (AP 12009) which was designed for the specific inhibition of TGF-beta2 biosynthesis. In vitro and in vivo experiments confirmed the mode of action, efficacy and tolerability of trabedersen and paved the way for clinical studies. In patients with high-grade glioma, intratumoral treatment with trabedersen is currently evaluated in a pivotal, randomized and active-controlled phase III study. Intravenous application of trabedersen for the treatment of patients with advanced pancreatic carcinoma, metastasizing melanoma, or metastatic colorectal carcinoma is assessed in a currently ongoing phase I/II dose escalation study. PMID- 21619537 TI - Biology of transforming growth factor-beta signaling. AB - Transforming growth factor (TGF)-beta signaling has been implicated as an important regulator of almost all major cell behaviors, including proliferation, differentiation, cell death, and motility. Which cell responses are induced or suppressed in response to TGF-beta depends on the cell type and the context in which TGF-beta signaling is received. TGF-beta ligands, their receptors, and intracellular Smad effectors lie in the center of TGF-beta signaling. TGF-beta ligands signal via receptor serine/threonine kinases that phosphorylate and activate Smad proteins as well as other signaling molecules. Smad complexes associate with chromatin and regulate transcription, defining the biological response of a cell to TGF-beta stimulation. In addition, numerous factors constitute complex networks to regulate TGF-beta signaling and to provide this cytokine with the ability to induce cellular context-specific cell responses. Perturbation of the network is strongly involved in various pathological situations, including cancer and fibrosis. In this review, we consider the basic machinery of TGF-beta signaling and describe several factors which make up TGF beta signaling networks. We also address major TGF-beta-induced cell responses involved in several physiological and pathological conditions, including cell proliferation, fibrosis, and epithelial-mesenchymal transition. PMID- 21619538 TI - TGF-beta2 signaling in high-grade gliomas. AB - High-grade gliomas are the most common primary tumors in the central nervous system (CNS) in adults. Despite efforts to improve treatment by combination therapies (neurosurgery, radio- and chemotherapy), high-grade glioma patients still have a grim prognosis, indicating an urgent need for new therapeutic approaches. The molecular processes of gliomagenesis are being unraveled, and novel targeted therapeutic strategies to defy high-grade gliomas are emerging. Transforming growth factor-beta (TGF-beta), in particular the TGF-beta2 isoform, has been identified as a key factor in the progression of malignant gliomas. TGF beta2, originally described as "glioblastoma-derived T-cell suppressor factor", is associated with the immuno-suppressed status of patients with glioblastoma, and is therefore responsible for loss of tumor immune surveillance. Elevated TGF beta2 levels in tumors and in the plasma of patients have been associated with advanced disease stage and poor prognosis. Consequently, a targeted strategy to modulate TGF-beta2 signaling is highly promising. The antisense oligonucleotide trabedersen (AP 12009) that specifically blocks TGF-beta2 mRNA will be the main focus of this review. In three phase I/II studies and a randomized, active controlled dose-finding phase IIb study, trabedersen treatment of high-grade glioma patients with recurrent or refractory tumor disease led to long-lasting tumor responses and so far promising survival data. On the basis of these data the currently ongoing phase III study SAPHIRRE was initiated. PMID- 21619539 TI - Therapeutic strategies to target TGF-beta in the treatment of bone metastases. AB - Bone is one of the most common organs to be affected in patients with metastatic cancer. These bone metastases are often accompanied by bone destruction, bone fractures, pain, and hypercalcemia. Transforming growth factor-beta (TGF-beta) is a major bone-derived factor that is released in active form upon osteoclastic bone resorption. TGF-beta, in turn, stimulates bone metastatic cells to secrete factors that further drive osteolytic destruction of the bone adjacent to the tumor, categorizing TGF-beta as a crucial factor responsible for driving the feed forward vicious cycle of cancer growth in bone. Moreover, TGF-beta activates epithelial-to-mesenchymal transition, increases tumor cell invasiveness and angiogenesis and induces immunosuppression. Blocking the TGF-beta signaling pathway to interrupt this vicious cycle between tumor cells and bone offers a promising target for therapeutic intervention to decrease skeletal metastasis. In this review, preclinical and clinical data are evaluated for the potential use of TGF-beta inhibitors in clinical practice to treat bone metastases. PMID- 21619540 TI - Structures of TGF-beta receptor complexes: implications for function and therapeutic intervention using ligand traps. AB - TGF-beta isoforms (TGF-beta1, -beta2, and -beta3) are secreted signaling ligands that stimulate the expression of protein components of the extracellular matrix, regulate the growth and differentiation of epithelial cells, modulate immune cell function, and play roles in the development of several essential organs, including the heart and lungs. The importance of the TGF-betas is underscored by their conservation among vertebrates and by their demonstrated roles in a variety of human diseases, including tissue fibrosis and cancer. The objective of this review is to highlight recent progress in characterizing the structures of the three TGF-beta isoforms in complex with their receptors, and to compare these with one another as well as with other members of the superfamily. Although the structural information and accompanying biophysical studies emphasize the shared ancestry of TGF-betas, they also provide insight as to how the TGF-betas diverged from other members of the superfamily and one another to fulfill distinct roles in vivo. The similarities and differences by which the isoforms bind their receptors present unique opportunities for designing pan-isoform and isoform specific ligand traps and progress toward developing these is described. PMID- 21619541 TI - Tgf-beta type I receptor (Alk5) kinase inhibitors in oncology. AB - The TGFbeta type I receptor kinase (ALK5) is an attractive target for intervention in TGFbeta signaling due to its druggability as well as its centrality and specificity in the pathway. A number of potent, selective ALK5 inhibitors have been discovered which interact with the ATP-binding site of ALK5. Crystallographic studies of these molecules bound to ALK5 have provided an understanding of potency and selectivity achieved by these inhibitors. ALK5 kinase inhibitors are potently active in models of cancer due to mechanisms of action similar to those for other TGFbeta inhibitory agents. Recent insights into the function of TGFbeta in human tumors as well as in preclinical models of cancer are helping to identify potential target patient populations and drug combinations for the development of ALK5 kinase inhibitors and other TGFbeta- targeted therapeutics. Differences in the toxicological effects, pharmacokinetics and clinical side effects of ALK5 kinase inhibitors and other TGFbeta-targeted agents provide a useful and differentiated set of TGFbeta signaling inhibitory agents to investigate in clinical studies. PMID- 21619542 TI - Role of TGF-beta in melanoma. AB - Human malignant melanoma is highly resistant to chemotherapy and current immunotherapeutic approaches induce long term remission only in the minority of patients. The transforming growth factor-beta (TGF-beta) has attracted much attention as a therapeutic target because it plays an important and pleiotropic role in melanoma progression. TGF-beta is a multifunctional cytokine involved in the regulation of many cellular processes including cell proliferation, differentiation and survival. Resistance to the growth inhibitory effects of TGF beta without alterations of TGF-beta signaling molecules is characteristic of cutaneous melanoma. Melanoma produces increasing amounts of TGF-beta with disease progression, inhibiting immune responses and providing an optimal microenvironment for undisturbed tumor growth. In addition, TGF-beta exerts its tumor promoting functions via direct effects on tumor cell motility and invasiveness and indirectly by modulating tumor stroma and extracellular matrix, supporting angiogenesis and inhibiting immune surveillance. TGF-beta acts through multiple intracellular signaling pathways and the outcome of TGF-beta signaling is context-dependent. Defining the impact of the different TGF-beta signaling pathways on melanoma progression will help to identify suitable therapeutic targets. Here we review the current knowledge of TGF-beta in melanoma and discuss recent therapeutic approaches targeting the TGF-beta pathway. PMID- 21619544 TI - TGF-beta as target in oncology. PMID- 21619543 TI - The complexities of TGF-beta action during mammary and squamous cell carcinogenesis. AB - Many advanced tumors produce excess amounts of Transforming Growth Factor-beta (TGF-beta), which is a potent growth inhibitor of normal epithelial cells. However, in tumors its homeostatic action on cells can be diverted along several alternative pathways. Thus, TGF-beta signaling has been reported to elicit a preventative or tumor suppressive effect during the earlier stages of tumorigenesis, but later in tumor development, when carcinoma cells become refractory to TGF-beta-mediated growth inhibition, response to TGF-beta signaling elicits predominantly tumor progressing effects. This is not a simple switch from suppression to progression, but more like a rheostat, involving multiple complementary and antagonizing activities that slowly tip the balance from one to the other. This review will focus on the multiple activities of TGF-beta in regulation of two epithelial tumor types, namely squamous cell carcinoma and breast cancer. Basic findings in current mouse models of cancer are presented, as well as a discussion of the complicating issue of outcome of altered TGFbeta signaling depending on genetic variability between mouse strains. This review also discusses the role TGF-beta within the tumor microenvironment particularly its ability to polarize the microenvironment towards a pro-tumorigenic milieu. PMID- 21619545 TI - The polyamine pathway as a potential target for vascular diseases: focus on restenosis. AB - Polyamines are organic polycations expressed by all living organisms, which are known to play an essential role in cell proliferation and differentiation. Recent studies revealed their involvement also in cell contractility and migration and in programmed cell death. These processes are known to contribute to restenosis, a pathophysiological process occurring in 10-20% of patients submitted to revascularization procedures. The advent of bare metal stents and of drug-eluting stents has significantly reduced but not eliminated the incidence of restenosis, which thus remains a clinically relevant problem. Despite the potential role of the polyamine pathway as a therapeutic target due to its involvement in proliferation, apoptosis and migration of vascular cells, experimental inhibition of polyamine synthesis and/or uptake has been poorly investigated in animal models of vascular disease. Here we review the current knowledge about molecular mechanisms related to polyamine functions, with particular reference to the role played by polyamines in vascular cell pathophysiology, together with experimental evidence obtained so far in animal models of (re)stenosis. We also evaluate the advantages of different routes of administration of polyamine synthesis/transport inhibitors and polyamine analogue molecules. Increasing knowledge about the molecular mechanisms and functions of polyamines is expected to shed new light on their potential role as a therapeutic target for restenosis reduction. PMID- 21619546 TI - MicroRNAs as potential therapeutic agents in the treatment of myocardial infarction. AB - In spite of enormous efforts, myocardial infarction is one of the most common causes of morbidity and mortality worldwide. The molecular mechanisms underlying the pathological myocardial alterations in affected patients are not fully elucidated. Recent studies have uncovered an important regulatory role for microRNAs (miRNAs), a family of small non-coding RNA molecules which - by translational repression or messenger RNA (mRNA) degradation - primarily act as negative regulators of gene expression. MiRNAs have been identified as regulatory key players during cellular differentiation, proliferation, and apoptosis. Recent work has unveiled an important impact of miRNAs on the pathophysiology of myocardial infarction and consecutive myocardial alterations, including arrhythmia, remodelling processes, cardiac fibrosis, and hypertrophia. Additionally, specific miRNAs have been identified to be either elevated or decreased in the blood plasma after myocardial infarction. Determination of miRNA expression levels therefore offers a potential prognostic and/or predictive value. Future therapeutic concepts aiming at attenuation of ischemia-induced harm and reduction of maladaptive changes may include strategies to influence altered miRNA expression patterns. In this review, we summarize current knowledge of the modulating role of miRNAs in pathological alterations occurring in myocardial infarction as well as currently available data concerning miRNAs as diagnostic markers and therapeutic targets. PMID- 21619547 TI - Recent advances and considerations for the future in forensic analysis of degraded DNA by autosomic miniSTR multiplex genotyping. AB - STR genotyping from degraded DNA samples requires genetic profiles to be obtained from DNA fragments no bigger than 200-300 bp. It requires the use of miniSTRs, which are smaller than the STRs used in standard typing. This paper reviews recent advances in miniSTR genotyping, beginning with a brief introduction to the processes involved in DNA fragmentation and how it hinders standard STR genotyping before proceeding further to the loci included in the main DNA databases and finishing with the International Workgroups' recommended design strategies for developing miniSTR reactions. The results of the efforts of many laboratories achieving different STR multiplexes and patents are also described and compared. Finally, a consideration of the perspectives for the future in this area is presented. PMID- 21619548 TI - Recent progresses in application of fullerenes in cosmetics. AB - Cosmetic industry is a fast growing industry with the continuous development of new active ingredients for skin care products. Fullerene C(60) and its derivates have been subject of intensive research in the last few years. Fullerenes display a wide range of different biological activities. Strong antioxidant capacities and effective quenching radical oxygen species (ROS) made fullerenes suitable active compounds in the formulation of skin care products. Published evidence on biological activities of fullerenes relevant for their application in cosmetics use and examples of published patents are presented. Recent trends in the use of fullerenes in topical formulations and patents are reviewed. Future investigations covering application of fullerenes in skin care are discussed. PMID- 21619549 TI - Treatment of effluents polluted by nitrogen with new biological technologies based on autotrophic nitrification-denitrification processes. AB - In recent years, various technologies have been developed for the removal of nitrogen from wastewater that is rich in nitrogen but poor in organic carbon, such as the effluents from anaerobic digesters and from certain industries. These technologies have resulted in several patents. The core of these technologies is some of the processes and patents described in this paper: Aerobic denitrification, Sharon, Anammox, OLAND, CANON, NOx process, DEMON. More specifically, one of the first innovative options described for removing nitrogen include partial nitrification under aerobic conditions (partial Sharon process) followed by autotrophic anaerobic oxidation (Anammox process). The partial Sharon Anammox process can be performed under alternating oxic and anoxic conditions in the same bioreactor or in two steps in two separate bioreactors. This overview focuses on the technical and biological aspects of these new types of treatment system, and compares them to other technologies. Given the fact that nitrification is a sensitive process, special attention is paid to conditions such as temperature, dissolved oxygen, hydraulic retention time, free ammonia, nitrous acid concentration, and pH. A discussion of the pros and cons of such treatment systems is also included since autotrophic nitrogen removal has advantages as well as drawbacks. The paper concludes with a discussion of future research that could improve these systems by enhancing performance and reducing costs. PMID- 21619550 TI - Farnesoid X receptor protects human and murine gastric epithelial cells against inflammation-induced damage. AB - Bile acids from duodenogastric reflux promote inflammation and increase the risk for gastro-oesophageal cancers. FXR (farnesoid X receptor/NR1H4) is a transcription factor regulated by bile acids such as CDCA (chenodeoxycholic acid). FXR protects the liver and the intestinal tract against bile acid overload; however, a functional role for FXR in the stomach has not been described. We detected FXR expression in the normal human stomach and in GC (gastric cancer). FXR mRNA and protein were also present in the human GC cell lines MKN45 and SNU5, but not in the AGS cell line. Transfection of FXR into AGS cells protected against TNFalpha (tumour necrosis factor alpha)-induced cell damage. We identified K13 (keratin 13), an anti-apoptotic protein of desmosomes, as a novel CDCA-regulated FXR-target gene. FXR bound to a conserved regulatory element in the proximal human K13 promoter. Gastric expression of K13 mRNA was increased in an FXR-dependent manner by a chow diet enriched with 1% (w/w) CDCA and by indomethacin (35 mg/kg of body weight intraperitoneal) in C57BL/6 mice. FXR-deficient mice were more susceptible to indomethacin-induced gastric ulceration than their WT (wild-type) littermates. These results suggest that FXR increases the resistance of human and murine gastric epithelial cells to inflammation-mediated damage and may thus participate in the development of GC. PMID- 21619551 TI - The combined effect of electrical stimulation and resistance isometric contraction on muscle atrophy in rat tibialis anterior muscle. AB - Electrical stimulation has been used to prevent muscle atrophy, but this method is different in many previous studies, appropriate stimulation protocol is still not decided. Although resistance exercise has also been shown to be an effective countermeasure on muscle atrophy, almost previous studies carried out an electrical stimulation without resistance. It was hypothesized that electrical stimulation without resistance is insufficient to contract skeletal muscle forcefully, and the combination of electrical stimulation and forceful resistance contraction is more effective than electrical stimulation without resistance to attenuate muscle atrophy. This study investigated the combined effects of electrical stimulation and resistance isometric contraction on muscle atrophy in the rat tibialis anterior muscle. The animals were divided into control, hindlimb unloading (HU), hindlimb unloading plus electrical stimulation (ES), and hindlimb unloading plus the combination of electrical stimulation and resistance isometric contraction (ES+IC). Electrical stimulation was applied to the tibialis anterior muscle percutaneously for total 240 sec per day. In the ES+IC group, the ankle joint was fixed to produce resistance isometric contraction during electrical stimulation. After 7 days, the cross-sectional areas of each muscle fiber type in the HU group decreased. Those were prevented in the ES+IC group rather than the ES group. The expression of heat shock protein 72 was enhanced in the ES and ES+IC groups. These results indicated that although electrical stimulation is effective to prevent muscle atrophy, the combination of electrical stimulation and isometric contraction have further effect. PMID- 21619552 TI - The significance of MGMT protein detection in evaluation of osteosarcoma necrosis rate after cisplatin chemotherapy. AB - The aim of this article was to investigate the correlation between expression of O(6)-methylguanine-DNA methyltransferase (MGMT) in osteosarcoma and the curative effect of alkylating agent (Cis-diaminodichloroplatinum, CDDP). 42 male patients and 34 female patients with a median age of 17 years (9 to 43 years) were eligible for this study. According to histopathological types, there were 3 cases of telangiectatic osteogenic sarcoma, 22 cases of osteoblastic, 11 cases of chondroblastic and 16 cases of fibroblastic sarcoma. Immunohistochemical method was used to detect the expression of MGMT protein. The correlations between MGMT expression and the curative effect of CDDP on osteosarcoma have been investigated. It was shown by immunohistochemical staining that among 76 osteosarcoma biopsy specimens, 52 (68%) cases were positive, 27 (35%) cases were weak positive, 18 (24%), cases were moderate positive, and 7 (9%) cases were strong positive. There were no significant differences in MGMT expression among different pathological types of tumors (p>0.5). After CDDP chemotherapy, among pathologic specimens in which MGMT expression was positive, necrosis rates were as follows: grade I, 5 cases (38%); grade II, 7 cases (25%); grade III, 15 cases (21%); grade IV, 2 cases (23%). Osteosarcoma necrosis rate was low when the expression of MGMT protein was positive, whereas necrosis rate was high when there was a low level of MGMT expression (p<0.01). There was a significant negative correlation between the level of MGMT expression in osteosarcoma tissue and osteosarcoma necrosis rate after cisplatin chemotherapy. PMID- 21619553 TI - Contribution of Ras farnesyl transferase, MAP kinase and cytochrome P-450 metabolites to endothelin-1 induced hypertension. AB - Endothelin 1 (ET-1) is vasoactive peptide that acts via ET-A receptors coupling inducing vascular smooth muscle cell proliferation and contraction. ET-1 is involved in the development and maintenance of hypertension. Aim of this study was to determine the contribution of Ras farnesyl transferase, mitogen activated protein kinase (MAP kinase) and cytochrome P!450 (CYP450) metabolites to ET-1 induced hypertension. ET-1 (5 pmol/kg per minute) was chronically infused into to the jugular vein by use of mini-osmotic pump for 9 days in male Sprague-Dawley rats. Mean arterial blood pressure (MABP) in ET-1-treated rats was 154+/-2 mm Hg (hypertensive rats) compared with 98+/-3 mm Hg in control (normotensive) rats. Infusion of Ras farnesyl transferase inhibitor FPTIII (138 ng/min), MAP kinase inhibitor PD-98059 (694 ng/min) and CYP450 inhibitor 17-ODYA (189 ng/min) significantly attenuated MABP to 115+/-2.5 mm Hg, 109+/-3 mm Hg and 118+/-1.5 mm Hg, respectively. These results suggest that CYP-450 metabolites and Ras/MAP kinase pathway contribute to the development of ET-1 induced hypertension. Further investigation has to be done to confirm whether activation of RAS/MAP kinase pathway by arachidonic acid metabolites plays an important role in the development of ET-1 induced hypertension. PMID- 21619554 TI - Comparative study of three rat models of stress urinary incontinence. AB - The aim of this study was to establish three rat models of stress urinary incontinence (SUI) and investigate the differences in urodynamics and success rate of establishing each model. Sixty healthy female SD rats were randomly divided into 4 groups of 15 rats each: A: control group; B: urethrolysis group; C: ovariectomy with colpectasis group; and D: colpectasis only group. Abdominal leak point pressure (LPP) and maximum bladder capacity (MBC) were calculated. The sneeze test was used to examine whether or not liquid leakage occurred at the urethral meatus in each model. LPP and MBC values were significantly lower in all experimental groups (groups B, C, and D) when compared to the control group A (p < 0.001). Within the experimental groups, the MBC and LPP of group B was the lowest, while these values in groups C and D were relatively higher (p < 0.001). However, there was no significant difference in LPP between groups C and D (p > 0.05). The positive rates of the sneeze test in groups A, B, C, and D were 0%, 86%, 64%, and 40%, respectively. We observed significant differences between the experimental groups and the control group (p < 0.05). Three methods (urethrolysis, colpectasis with an ovariectomy, and colpectasis only) can be successfully used to establish models of SUI in rats. Significant differences in urodynamics and the successful establishment of the model occurred in this study. PMID- 21619555 TI - Antimicrobial resistance status and prevalence rates of extended spectrum beta lactamase (ESBL) producers isolated from a mixed human population. AB - Owing to the increasing epidemiological and therapeutic challenges associated with infections due to ESBL producers, ESBL prevalence rate among some bacteria isolates from healthy and non-healthy human population in a metropolitan Nigerian setting was evaluated. A total of one hundred and forty-five (145) bacteria strains were isolated from a total of four hundred and sixty (460) samples collected from urine, wound, throat and anal swabs of 220 healthy volunteers in the community and from 240 patients in 2 secondary and 2 tertiary hospitals (altogether, 4) in Enugu metropolis. The presumptive confirmatory test used for ESBL detection was the Double Disc Synergy Test (DDST) method. Conjugation and plasmid curing studies were also done for resistance factor determination. Of the 145 isolates, 20 were ESBL producers with 35% of these ESBL producers being of community origin and 65% from hospitals. This translates to 4.8% and 9% incidences (comparably higher than established prevalence of 4.4% and 7.5 respectively) for community and hospital infections respectively. The ESBL isolates showed high resistance to tetracycline, gentamicin, pefloxacin, ceftriaxone, cefuroxime, ciprofloxacin and Augmentin((r)) (Amoxicilin and clavulanic acid combination). Conjugation studies for Resistance plasmid transfer showed non-transference of resistance determinants between the ESBL transconjugants and recipient strains. Correspondingly, the plasmid curing studies revealed that the acridine orange could not effect a cure on the isolates as they still retained high resistance to the antibiotics after the treatment. This study confirms the growing incidences/pool of ESBL strains in Nigeria and call for widespread and continuous monitoring towards an effective management of the potential therapeutic hurdle posed by this trend. PMID- 21619556 TI - Outpatient antihypertensive drug utilization in Canton Sarajevo during five years period (2004-2008) and adherence to treatment guidelines assessment. AB - Hypertension is chronic disease with high prevalence, which can successfully be treated with antihypertensive drugs. Previous researches have shown that existing hypertension treatment guidelines are not fully implemented in practice. We have analysed antihypertensive drug utilization in Canton Sarajevo during five-year period (2004-2008). Research findings are discussed in relation to expected drug utilization according to Canton Sarajevo treatment guidelines. Objective of this research is to examine prescription patterns of antihypertensive drugs in primary health care in Canton Sarajevo during five-year period. Based on study findings we did an estimation of adherence to local treatment guidelines, which are similar to those published globally. Drug utilization data were collected from the largest pharmacy (retail) chain, representing more than 80% of pharmacies in Canton Sarajevo. Following drug groups have been analyzed: diuretics, beta blockers, calcium-channel-blockers, ACE-Inhibitors (plain and combinations), Angiotensin-II-antagonists and alpha-blockers. Drug utilization is expressed in number of defined daily dose (DDDs), defined daily dose per thousand inhabitants per day (DDD/TID), drug utilization 90% (DU90%) and value in euros. ACE Inhibitors are most prescribed drug class; combination of ACE-Inhibitors and diuretics account 46% of total antihypertensive budget spending. ACEIs are followed by calcium-channel-blockers. Diuretics utilization is decreasing from 2006 and being replaced with beta-blockers. Diuretics, recommended as first line therapy, are ranked as third in total antihypertensive drug utilization. It is necessary to introduce follow-up and enforce adherence to developed treatment guideline. Drug utilization studies can be used as tool for assessment of treatment guidelines adherence in primary health care. PMID- 21619557 TI - Intra-amniotic administration of exogenous pulmonary surfactant for improving in lung maturity of fetal rabbits with intrauterine infection caused by premature rupture of membranes. AB - This study was to investigate the effect of intra-amniotic administration of pulmonary surfactant (PS) on lung maturation in conditions of experimentally induced premature rupture of membranes (PROM) and intrauterine infection of rabbits. To establish animal (rabbit) models of intrauterine infection caused by PROM, E. coli was intrauterinely injected in 24-and 26-day pregnant animals. Twenty healthy pregnancy adult Japanese white rabbits were divided into three groups: the infection group (8 rabbits), infection group with intra-amniotic PS administration (8 rabbits) and healthy controls (4 rabbits). Ultrastructure changes in the lung were observed under an electron microscope 19.5 hours after intervention. The results showed that the lung infection levels of fetal rabbits in the infection group and the infection group with PS had no significant difference. Bacillus bodies and infiltrated inflammatory cells can be occasionally seen in the alveoli, bronchial lumen, and cytoplasm.The type II alveolar epithelial cells (AT-II) were decreased in the infection group than that in control group. Lamellar bodies showed vacuolation changes and different levels of apoptosis. In the infection group with PS, the count of AT-II did not show an obvious decrease. Lamellar bodies increased, and different secretion forms appeared. Also, there was little apoptosis and no obvious collagen fiber hyperplasia in antenatal administration of PS group. We believe that once PROM has happened, intrauterine infection and inflammation stimulated a cascade reaction on the fetal lung, leading to abnormal changes in the alveolar ultrastructure. Intra-amniotic administration of PS can improve the fetal lung ultrastructure and its maturity. PMID- 21619558 TI - The effects of hypoglycemic and alcoholic coma on the blood-brain barrier permeability. AB - In this investigation, the effects of hypoglycemic coma and alcoholic coma on the blood-brain barrier (BBB) permeability have been compared. Female adult Wistar albino rats weighing 180-230 g were divided into three groups: Control group (n=8), Alcoholic Coma Group (n=18), and Hypoglycemic Coma group (n=12). The animals went into coma approximately 3-4 hours after insulin administration and 3 5 minutes after alcohol administration. Evans blue (4mL/kg) was injected intravenously as BBB tracer. It was observed that the alcoholic coma did not significantly increase the BBB permeability in any of the brain regions when compared to control group. Changes in BBB permeability were significantly increased by the hypoglycemic coma in comparison to the control group values (p<0.01). Our findings suggest that hypoglycemic and alcoholic coma have different effects on the BBB permeability depending on the energy metabolism. PMID- 21619559 TI - Preemptive analgesic effects of midazolam and diclofenac in rat model. AB - The aim of the present study was to investigate the preemptive analgesic effects of intraperitoneally administrated midazolam and diclofenac, before acute and inflammatory induced pain in rat model. One hundred twenty-eight (n=8 in each group) male Sprague Dawley rats were included in the study. Paw movements in response to thermal stimulation or paw flinching in response to formalin injection were compared after midazolam (0.1, 1, 5 and 10 mg/kg) and diclofenac (10 mg/kg), intraperitoneal administration. Saline was used as a control. Preemptive analgesic effect was significant in both tests when diclofenac and midazolam was administrated before the pain stimuli (p<0.01 and p<0.001). Intraperitoneal injection of midazolam in doses 5 and 10 mg/kg, increase the response time in hot plate test and decrease the number of flinches in formalin test (p<0.01 vs. p<0.001). ED50 of midazolam (with diclofenac) in hot plate test was 2.02 mg/kg (CI95% =-3.47-5.03 mg); and, 0.9 mg/kg (CI95% =-0.87-4.09 mg) in phase I and 0.7 mg/kg (CI95% = 0.48-6.63 mg) in phase II, in formalin test.Intraperitoneally administered midazolam and diclofenac had preemptive analgesic effects on acute thermal, and inflammatory induced pain in rats. PMID- 21619560 TI - Skin and kidney histological changes in graft-versus-host disease (GVHD) after kidney transplantation. AB - Kidney transplantation (Ktx) is generally performed during end stage renal disease due to a loss of the kidneys' ability to filter wastes from the circulatory system. Acute graft-versus-host disease (GVHD) after Ktx is a life threatening complication that progresses to organ failure, systemic complications, and death. The current study evaluated the significance of histologic findings of GVHD as obtained from skin biopsies following Ktx in swine. A swine model of Ktx with tacrolimus-based immunosuppression was used to assess possible correlations between acute-graft-cellular rejection and skin histological findings for prediction of GVHD. Animals were divided into a Ktx treatment group or a control group with no Ktx and skin and kidney biopsies were histologically assessed at postoperative days 0, 15, 30, 45 and 60. Skin samples were analyzed and classified from grade 1 to 4 of skin GVHD and the major histopathological changes of kidney acute cellular rejection were described using Banff's score system. We observed a significant linear correlation between the histological grading values of skin biopsy changes and the histological grading values of kidney biopsies (Kendall's tau_b=0.993) in the Ktx experimental group. No histological changes were observed in controls. Our findings demonstrate the diagnostic value of staging skin GVHD after Ktx and suggest it's future utility for monitoring long term Ktx-induced changes. PMID- 21619561 TI - The tissue velocity imaging and strain rate imaging in the assessment of interatrial electromechanical conduction in patients with sick sinus syndrome before and after pacemaker implantation. AB - Tissue velocity imaging (TVI) and strain rate imaging (SRI) were recently introduced to quantify myocardial mechanical activity in patients receiving cardiac resynchronization therapy. To clear whether atrial-demand-based (AAI) (R) atrial pacing can fully simulate the electromechanical conduction of physiological state and to clarify which one is more appropriate for the assessment of electromechanical activity of the heart between TVI and SRI, 30 normal subjects and 31 patients with sick sinus syndrome (SSS) before and after AAI(R) pacemaker implantation (PI) were investigated in this study. The results showed that the time intervals (ms), P-SRa assessed by SRI (not P-Va assessed by TVI) prolonged step by step from the lateral wall of the right atrium (RA), the interatrial septum (IAS) and the left atrium (LA) in normal subjects(5.01+/-0.62, 17.05+/-3.54 and 45.09+/-12.26, p<0.01). P-Va and P-SRa did not differ at the RA, IAS and LA in patients with SSS before PI (p>0.05), and they were significant longer than those of normal subjects (p<0.01). However, they shortened to normal levels in patients with SSS after PI and P-SRa showed again the trend of gradually prolonging from the RA, IAS to LA. At the same time, the peak velocities and the peak strain rates during atrial contraction also returned to normal values from lower levels. These data suggested that AAI(R) atrial pacing can successfully reverse the abnormal interatrial electromechanical conduction in patients with SSS, and SRI is more appropriate for the assessment of the electromechanical activity of atrial wall than TVI. PMID- 21619562 TI - Bilateral retinal detachment in a case of preeclampsia. AB - Serous retinal detachment is an unusual cause of visual loss in preeclampsia. We report the case of 24-years-old primipara with severe preeclampsia who developed bilateral serous retinal detachment a few hours after delivery. A few weeks after delivery there was spontaneous resorption of the subretinal fluid and complete resolution bilateral serous retinal detachment, with residual pigmentary changes of the retinal pigment epithelium. Visual acuity was normal in each eye. The management of retinal detachment as a complication in preeclampsia is conservative and the prognosis is usually good. PMID- 21619563 TI - Angiosarcoma of the ovary in an 11 year old girl: case report and review of the literature. AB - Sarcomas of the female genital tract in general are rare and ovarian sarcomas comprise less than 1% of ovarian malignancies. In the literature there are 15 reported angiosarcomas of patients 21 year old and younger with no one originated in the ovary. We report a case of ovarian angiosarcoma in an 11 year old girl, presented with left side hip pain. MRI of abdomen and pelvis confirmed expansive solid and cystic mass occupied both ovaries. Imunohistochemistry staining was performed, CD34, Factor VIII, CD31, in order to confirm the diagnosis. Final diagnosis was angiosarcoma. The patient received 6 cycles of chemotherapy, according to the CWS-2002P protocol. 8 months after the diagnosis was established, there were no signs of any tumors according to the ultrasound, CT scan, and MRI. Although, extremely rare, angiosarcoma can also affect children and this diagnosis should be considered carefully in tumor with rich vascular network, necrosis and brisk mitotic activity. PMID- 21619564 TI - Collet Sicard syndrome as atypical presentation of neck fibrosarcoma: a case report. AB - We report a 57 years old female patient with neck fibrosarcoma. Her main complaints consisted of hoarseness, difficulty swallowing, pain in the left side of her neck and left shoulder region, which all indicated the Collet Sicard syndrome, so the working diagnosis was glomus tumor. Diagnostic MSCT was used, and the characteristics of the radiologic finding did not indicate any of the paraganglioma types, although the tumor was localized in the area of the carotid bifurcation, demonstrating the signs of extension into the jugular foramen. The patient has been treated surgically in general anesthesia and pathologic diagnosis was fibrosarcoma. PMID- 21619565 TI - Nurse prescribing of medicines in Western European and Anglo-Saxon countries: a systematic review of the literature. AB - BACKGROUND: A growing number of countries are introducing some form of nurse prescribing. However, international reviews concerning nurse prescribing are scarce and lack a systematic and theoretical approach. The aim of this review was twofold: firstly, to gain insight into the scientific and professional literature describing the extent to and the ways in which nurse prescribing has been realised or is being introduced in Western European and Anglo-Saxon countries; secondly, to identify possible mechanisms underlying the introduction and organisation of nurse prescribing on the basis of Abbott's theory on the division of professional labor. METHODS: A comprehensive search of six literature databases and seven websites was performed without any limitation as to date of publication, language or country. Additionally, experts in the field of nurse prescribing were consulted. A three stage inclusion process, consisting of initial sifting, more detailed selection and checking full-text publications, was performed independently by pairs of reviewers. Data were synthesized using narrative and tabular methods. RESULTS: One hundred and twenty-four publications met the inclusion criteria. So far, seven Western European and Anglo-Saxon countries have implemented nurse prescribing of medicines, viz., Australia, Canada, Ireland, New Zealand, Sweden, the UK and the USA. The Netherlands and Spain are in the process of introducing nurse prescribing. A diversity of external and internal forces has led to the introduction of nurse prescribing internationally. The legal, educational and organizational conditions under which nurses prescribe medicines vary considerably between countries; from situations where nurses prescribe independently to situations in which prescribing by nurses is only allowed under strict conditions and supervision of physicians. CONCLUSIONS: Differences between countries are reflected in the jurisdictional settlements between the nursing and medical professions concerning prescribing. In some countries, nurses share (full) jurisdiction with the medical profession, whereas in other countries nurses prescribe in a subordinate position. In most countries the jurisdiction over prescribing remains predominantly with the medical profession. There seems to be a mechanism linking the jurisdictional settlements between professions with the forces that led to the introduction of nurse prescribing. Forces focussing on efficiency appear to lead to more extensive prescribing rights. PMID- 21619566 TI - Gender differences and determinants of health related quality of life in coronary patients: a follow-up study. AB - BACKGROUND: The role of gender differences in Health Related Quality Life (HRQL) in coronary patients is controversial, so understanding the specific determinants of HRQL in men and women might be of clinical importance. The aim of this study was to know the gender differences in the evolution of HRQL at 3 and 6 months after a coronary event, and to identify the key clinical, demographic and psychological characteristics of each gender associated with these changes. METHODS: A follow-up study was carried out, and 175 patients (112 men and 63 women) with acute myocardial infarction (AMI) or unstable angina were studied. The SF-36v1 health questionnaire was used to assess HRQL, and the GHQ-28 (General Health Questionnaire) to measure mental health during follow-up. To study the variables related to changes in HRQL, generalized estimating equation (GEE) models were performed. RESULTS: Follow-up data were available for 55 men and 25 women at 3 months, and for 35 men and 12 women at 6 months. Observations included: a) Revascularization was performed later in women. b) The frequency of rehospitalization between months 3 and 6 of follow-up was higher in women c) Women had lower baseline scores in the SF-36. d) Men had progressed favourably in most of the physical dimensions of the SF-36 at 6 months, while at the same time women's scores had only improved for Physical Component Summary, Role Physical and Social Functioning; e) the variables determining the decrease in HRQL in men were: worse mental health and angina frequency; and in women: worse mental health, history of the disease, revascularization, and angina frequency. CONCLUSIONS: There are differences in the evolution of HRQL, between men and women after a coronary attack. Mental health is the determinant most frequently associated with HRQL in both genders. However, other clinical determinants of HRQL differed with gender, emphasizing the importance of individualizing the intervention and the content of rehabilitation programs. Likewise, the recognition and treatment of mental disorders in these patients could be crucial. PMID- 21619567 TI - Basing care reforms on evidence: the Kenya health sector costing model. AB - BACKGROUND: The Government of the Republic of Kenya is in the process of implementing health care reforms. However, poor knowledge about costs of health care services is perceived as a major obstacle towards evidence-based, effective and efficient health care reforms. Against this background, the Ministry of Health of Kenya in cooperation with its development partners conducted a comprehensive costing exercise and subsequently developed the Kenya Health Sector Costing Model in order to fill this data gap. METHODS: Based on standard methodology of costing of health care services in developing countries, standard questionnaires and analyses were employed in 207 health care facilities representing different trustees (e.g. Government, Faith Based/Nongovernmental, private-for-profit organisations), levels of care and regions (urban, rural). In addition, a total of 1369 patients were randomly selected and asked about their demand-sided costs. A standard step-down costing methodology was applied to calculate the costs per service unit and per diagnosis of the financial year 2006/2007. RESULTS: The total costs of essential health care services in Kenya were calculated as 690 million Euros or 18.65 Euro per capita. 54% were incurred by public sector facilities, 17% by Faith Based and other Nongovernmental facilities and 23% in the private sector. Some 6% of the total cost is due to the overall administration provided directly by the Ministry and its decentralised organs. Around 37% of this cost is absorbed by salaries and 22% by drugs and medical supplies. Generally, costs of lower levels of care are lower than of higher levels, but health centres are an exemption. They have higher costs per service unit than district hospitals. CONCLUSIONS: The results of this study signify that the costs of health care services are quite high compared with the Kenyan domestic product, but a major share are fixed costs so that an increasing coverage does not necessarily increase the health care costs proportionally. Instead, productivity will rise in particular in under-utilized private health care institutions. The results of this study also show that private-for-profit health care facilities are not only the luxurious providers catering exclusively for the rich but also play an important role in the service provision for the poorer population. The study findings also demonstrated a high degree of cost variability across private providers, suggesting differences in quality and efficiencies. PMID- 21619568 TI - A prospective cohort study of neighborhood stress and ischemic heart disease in Japan: a multilevel analysis using the JACC study data. AB - BACKGROUND: A body of research has shown that neighborhood environment may have an effect on a variety of health outcomes, including cardiovascular disease. One explanation for the mechanism of the effect of neighborhood on cardiovascular disease is psychosocial pathways. Direct evidence for an effect of neighborhood on cardiovascular disease with adjustment for perceived stress at the individual level has not been obtained, however. The Japan Collaborative Cohort Study for the Evaluation of Cancer Risk provides a unique dataset which has aggregated area based cohorts from 45 areas throughout Japan. The purpose of the present study was to examine the contextual effect of area-level stress on ischemic heart disease using data from a large prospective cohort in Japan. METHODS: A baseline survey of 110,792 residents of 45 areas aged 40-79 years was conducted between 1988 and 1990. Analysis was restricted to subjects from the 33 of 45 areas providing information about self-rated stress (32183 men and 45896 women). Multilevel Poisson regression models were employed in a two-level structure of individuals nested within the 33 areas. Area-level stress was calculated by sex as the number of persons who rated their stress level as high divided by the total number of subjects in that area. Mortality rate ratios (MRRs) per 1 percentage point increase in area-level stress were estimated with adjustment for compositional individual factors. RESULTS: During 15 years of follow-up (1,116,895 person-years), 936 deaths due to ischemic heart disease were recorded. Area-level stress varied from 6% to 22%. In the multivariable models, MRRs of area-level stress were 1.06 (95% confidence interval: 1.00-1.12, p = 0.043) in men and 1.07 (95% confidence interval: 1.00-1.14, p = 0.057) in women. CONCLUSIONS: Area-level stress affects the likelihood of death due to ischemic heart disease of individuals in men. The present findings may suggest that stress should be considered not only within the individual but also within the neighborhood context. PMID- 21619569 TI - Context-dependent codon partition models provide significant increases in model fit in atpB and rbcL protein-coding genes. AB - BACKGROUND: Accurate modelling of substitution processes in protein-coding sequences is often hampered by the computational burdens associated with full codon models. Lately, codon partition models have been proposed as a viable alternative, mimicking the substitution behaviour of codon models at a low computational cost. Such codon partition models however impose independent evolution of the different codon positions, which is overly restrictive from a biological point of view. Given that empirical research has provided indications of context-dependent substitution patterns at four-fold degenerate sites, we take those indications into account in this paper. RESULTS: We present so-called context-dependent codon partition models to assess previous empirical claims that the evolution of four-fold degenerate sites is strongly dependent on the composition of its two flanking bases. To this end, we have estimated and compared various existing independent models, codon models, codon partition models and context-dependent codon partition models for the atpB and rbcL genes of the chloroplast genome, which are frequently used in plant systematics. Such context-dependent codon partition models employ a full dependency scheme for four fold degenerate sites, whilst maintaining the independence assumption for the first and second codon positions. CONCLUSIONS: We show that, both in the atpB and rbcL alignments of a collection of land plants, these context-dependent codon partition models significantly improve model fit over existing codon partition models. Using Bayes factors based on thermodynamic integration, we show that in both datasets the same context-dependent codon partition model yields the largest increase in model fit compared to an independent evolutionary model. Context dependent codon partition models hence perform closer to codon models, which remain the best performing models at a drastically increased computational cost, compared to codon partition models, but remain computationally interesting alternatives to codon models. Finally, we observe that the substitution patterns in both datasets are drastically different, leading to the conclusion that combined analysis of these two genes using a single model may not be advisable from a context-dependent point of view. PMID- 21619570 TI - Automatic identification of gait events using an instrumented sock. AB - BACKGROUND: Textile-based transducers are an emerging technology in which piezo resistive properties of materials are used to measure an applied strain. By incorporating these sensors into a sock, this technology offers the potential to detect critical events during the stance phase of the gait cycle. This could prove useful in several applications, such as functional electrical stimulation (FES) systems to assist gait. METHODS: We investigated the output of a knitted resistive strain sensor during walking and sought to determine the degree of similarity between the sensor output and the ankle angle in the sagittal plane. In addition, we investigated whether it would be possible to predict three key gait events, heel strike, heel lift and toe off, with a relatively straight forward algorithm. This worked by predicting gait events to occur at fixed time offsets from specific peaks in the sensor signal. RESULTS: Our results showed that, for all subjects, the sensor output exhibited the same general characteristics as the ankle joint angle. However, there were large between subjects differences in the degree of similarity between the two curves. Despite this variability, it was possible to accurately predict gait events using a simple algorithm. This algorithm displayed high levels of trial-to-trial repeatability. CONCLUSIONS: This study demonstrates the potential of using textile-based transducers in future devices that provide active gait assistance. PMID- 21619571 TI - Factors affecting the disclosure of diabetes by ethnic minority patients: a qualitative study among Surinamese in the Netherlands. AB - BACKGROUND: Diabetes and related complications are common among ethnic minority groups. Community-based social support interventions are considered promising for improving diabetes self-management. To access such interventions, patients need to disclose their diabetes to others. Research on the disclosure of diabetes in ethnic minority groups is limited. The aim of our study was to explore why diabetes patients from ethnic minority populations either share or do not share their condition with people in their wider social networks. METHODS: We conducted a qualitative study using semi-structured interviews with 32 Surinamese patients who were being treated for type 2 diabetes by general practitioners in Amsterdam, the Netherlands. RESULTS: Most patients disclosed their diabetes only to very close family members. The main factor inhibiting disclosure to people outside this group was the Surinamese cultural custom that talking about disease is taboo, as it may lead to shame, gossip, and social disgrace for the patient and their family. Nevertheless, some patients disclosed their diabetes to people outside their close family circles. Factors motivating this decision were mostly related to a need for facilities or support for diabetes self-management. CONCLUSIONS: Cultural customs inhibited Surinamese patients in disclosing their diabetes to people outside their very close family circles. This may influence their readiness to participate in community-based diabetes self-management programmes that involve other groups. What these findings highlight is that public health researchers and initiatives must identify and work with factors that influence the disclosure of diabetes if they are to develop community-based diabetes self-management interventions for ethnic minority populations. PMID- 21619572 TI - Patients as healthcare consumers in the public and private sectors: a qualitative study of acupuncture in the UK. AB - BACKGROUND: The aim of this study was to compare patients' experiences of public and private sector healthcare, using acupuncture as an example. In the UK, acupuncture is popular with patients, is recommended in official guidelines for low back pain, and is available in both the private sector and the public sector (NHS). Consumerism was used as a theoretical framework to explore patients' experiences. METHODS: Semi-structured face-to-face interviews were conducted in 2007-8 with a purposive sample of 27 patients who had recently used acupuncture for painful conditions in the private sector and/or in the NHS. Inductive thematic analysis was used to develop themes that summarised the bulk of the data and provided insights into consumerism in NHS- and private practice-based acupuncture. RESULTS: Five main themes were identified: value for money and willingness to pay; free and fair access; individualised holistic care: feeling cared for; consequences of choice: empowerment and vulnerability; and "just added extras": physical environment. Patients who had received acupuncture in the private sector constructed detailed accounts of the benefits of private care. Patients who had not received acupuncture in the private sector expected minimal differences from NHS care, and those differences were seen as not integral to treatment. The private sector facilitated consumerist behaviour to a greater extent than did the NHS, but private consumers appeared to base their decisions on unreliable and incomplete information. CONCLUSIONS: Patients used and experienced acupuncture differently in the NHS compared to the private sector. Eight different faces of consumerist behaviour were identified, but six were dominant: consumer as chooser, consumer as pragmatist, consumer as patient, consumer as earnest explorer, consumer as victim, and consumer as citizen. The decision to use acupuncture in either the private sector or the NHS was rarely well-informed: NHS and private patients both had misconceptions about acupuncture in the other sector. Future research should evaluate whether the differences we identified in patients' experiences across private and public healthcare are common, whether they translate into significant differences in clinical outcomes, and whether similar faces of consumerism characterise patients' experiences of other interventions in the private and public sectors. PMID- 21619573 TI - Limited access to HIV prevention in French prisons (ANRS PRI2DE): implications for public health and drug policy. AB - BACKGROUND: Overpopulation, poor hygiene and disease prevention conditions in prisons are major structural determinants of increased infectious risk within prison settings but evidence-based national and WHO guidelines provide clear indications on how to reduce this risk. We sought to estimate the level of infectious risk by measuring how French prisons adhere to national and WHO guidelines. METHODS: A nationwide survey targeting the heads of medical (all French prisons) and psychiatric (26 French prisons) units was conducted using a postal questionnaire and a phone interview mainly focusing on access to prevention interventions, i.e. bleach, opioid substitution treatment (OST), HBV vaccination and post-exposure prophylaxis (PEP) for French prisoners. Two scores were built reflecting adherence to national and WHO international guidelines, ranging from 0 (no adherence) to 10 (maximum adherence) and 0 to 9 respectively. RESULTS: A majority (N=113 (66%)) of the 171 prisons answered the questionnaires, representing 74% coverage (46,786 prisoners) of the French prison population: 108 were medical units and 12 were psychiatric units. Inmate access to prevention was poor. The median[IQR] score measuring adherence to national guidelines was quite low (4.5[2.5; 5.5]) but adherence to WHO guidelines was even lower 2.5[1.5; 3.5]; PEP was absent despite reported risky practices. Unsuitable OST delivery practices were frequently observed. CONCLUSIONS: A wide gap exists between HIV prevention policies and their application in prisons. Similar assessments in other countries may be needed to guide a global policy reform in prison settings. Adequate funding together with innovative interventions able to remove structural and ideological barriers to HIV prevention are now needed to motivate those in charge of prison health, to improve their working environment and to relieve French prisoners from their currently debilitating conditions. PMID- 21619574 TI - Effectiveness of automated locomotor training in patients with acute incomplete spinal cord injury: a randomized controlled multicenter trial. AB - BACKGROUND: A large proportion of patients with spinal cord injury (SCI) regain ambulatory function. However, during the first 3 months most of the patients are not able to walk unsupported. To enable ambulatory training at such an early stage the body weight is partially relieved and the leg movements are assisted by two therapists. A more recent approach is the application of robotic based assistance which allows for longer training duration. From motor learning science and studies including patients with stroke, it is known that training effects depend on the duration of the training. Longer trainings result in a better walking function. The aim of the present study is to evaluate if prolonged robot assisted walking training leads to a better walking outcome in patients with incomplete SCI and whether such training is feasible or has undesirable effects. METHODS/DESIGN: Patients from multiple sites with a subacute incomplete SCI and who are not able to walk independently will be randomized to either standard training (3-5 sessions per week, session duration maximum 25 minutes) or an intensive training (3-5 sessions per week, session duration minimum 50 minutes). After 8 weeks of training and 4 months later the walking ability, the occurrence of adverse events and the perceived rate of exertion as well as the patients' impression of change will be compared between groups. TRIAL REGISTRATION: This study is registered at clinicaltrials.gov, identifier: NCT01147185. PMID- 21619575 TI - Patient involvement in patient safety: Protocol for developing an intervention using patient reports of organisational safety and patient incident reporting. AB - BACKGROUND: Patients have the potential to provide a rich source of information on both organisational aspects of safety and patient safety incidents. This project aims to develop two patient safety interventions to promote organisational learning about safety - a patient measure of organisational safety (PMOS), and a patient incident reporting tool (PIRT) - to help the NHS prevent patient safety incidents by learning more about when and why they occur. METHODS: To develop the PMOS 1) literature will be reviewed to identify similar measures and key contributory factors to error; 2) four patient focus groups will ascertain practicality and feasibility; 3) 25 patient interviews will elicit approximately 60 items across 10 domains; 4) 10 patient and clinician interviews will test acceptability and understanding. Qualitative data will be analysed using thematic content analysis.To develop the PIRT 1) individual and then combined patient and clinician focus groups will provide guidance for the development of three potential reporting tools; 2) nine wards across three hospital directorates will pilot each of the tools for three months. The best performing tool will be identified from the frequency, volume and quality of reports. The validity of both measures will be tested. 300 patients will be asked to complete the PMOS and PIRT during their stay in hospital. A sub-sample (N = 50) will complete the PMOS again one week later. Health professionals in participating wards will also be asked to complete the AHRQ safety culture questionnaire. Case notes for all patients will be reviewed. The psychometric properties of the PMOS will be assessed and a final valid and reliable version developed. Concurrent validity for the PIRT will be assessed by comparing reported incidents with those identified from case note review and the existing staff reporting scheme. In a subsequent study these tools will be used to provide information to wards/units about their priorities for patient safety. A patient panel will provide steering to the research. DISCUSSION: The PMOS and PIRT aim to provide a reliable means of eliciting patient views about patient safety. Both interventions are likely to have relevance and practical utility for all NHS hospital trusts. PMID- 21619576 TI - A perspective on SIDS pathogenesis. the hypotheses: plausibility and evidence. AB - Several theories of the underlying mechanisms of Sudden Infant Death Syndrome (SIDS) have been proposed. These theories have born relatively narrow beach-head research programs attracting generous research funding sustained for many years at expense to the public purse. This perspective endeavors to critically examine the evidence and bases of these theories and determine their plausibility; and questions whether or not a safe and reasoned hypothesis lies at their foundation. The Opinion sets specific criteria by asking the following questions: 1. Does the hypothesis take into account the key pathological findings in SIDS? 2. Is the hypothesis congruent with the key epidemiological risk factors? 3. Does it link 1 and 2? Falling short of any one of these answers, by inference, would imply insufficient grounds for a sustainable hypothesis. Some of the hypotheses overlap, for instance, notional respiratory failure may encompass apnea, prone sleep position, and asphyxia which may be seen to be linked to co-sleeping. For the purposes of this paper, each element will be assessed on the above criteria. PMID- 21619577 TI - Association of beta3-adrenergic receptor (ADRB3) Trp64Arg gene polymorphism with obesity and metabolic syndrome in the Balinese: a pilot study. AB - BACKGROUND: Prevalence of obesity is increasing all over the world. ADRB3 Trp64Arg gene polymorphism was proposed to be associated with obesity, although inconsistent findings and differences of the Arg64 allele frequency among various ethnics were reported. Westernization was reported to increase the prevalence of obesity in developing world. In this study we determined the prevalence of obesity and metabolic syndrome among urban and rural Balinese, and studied the association of ADRB3 Trp64Arg polymorphism with obesity and MetS. FINDINGS: A total of 528 Balinese (urban 282, rural 246) were recruited. Body mass index (BMI) and waist circumference (WC) were determined; high-density lipoprotein cholesterol (HDL-C), triglyceride (TG), systolic and diastolic blood pressure (SBP and DBP), and fasting plasma glucose (FPG) were measured using standard procedures. BMI and WC classifications were based on WHO classifications for Asian. Metabolic syndrome (MetS) was defined as described in the Joint Interim Statement. Chi-square test was employed to test the association between the ADRB3 Trp64Arg genotype and disease traits.Urban have higher BMI (p = 2.8 * 10-13), WC ( p < 2.2 * 10-16), TG (p = 0.0028), DBP (p = 1.8 * 10-5), and lower HDL-C (p = 0.0376) when compared to rural. Abdominal obesity and MetS prevalence were significantly higher in urban as compared to rural (both p < 0.001). The Arg64 allele frequency was similar between urban (0.06) and rural (0.05). The Arg64 rural female carriers have higher BMI and WC as compared to their Trp64 counterparts (p = 0.041 for BMI and p = 0.012 for WC), and consequently higher abdominal obesity prevalence (p = 0.007). Comparison between male and female, as well as urban and rural, showed different prevalence of MetS co-morbidities. Abdominal obesity and hypertriglyceridaemia were consistently appeared in all groups, suggesting to play a role as determinant of MetS in both urban and rural. CONCLUSIONS: Prevalence of obesity and MetS in urban were two times higher when compared to rural. Abdominal obesity and hypertriglyceridaemia appears to be the key determinant of MetS in both urban and rural Balinese. Our results indicated an association of the ADRB3 Trp64Arg gene polymorphism with obesity in the rural female. PMID- 21619578 TI - Interleukin-11-induced capillary leak syndrome in primary hepatic carcinoma patients with thrombocytopenia. AB - BACKGROUND: Capillary leak syndrome (CLS) is a rare condition characterized by recurrent episodes of generalized edema and severe hypotension associated with hypoproteinemia. Interleukin-11 (IL-11) is a promising therapeutic agent for thrombocytopenia. A direct correlation between IL-11 and CLS has never been reported previously, particularly in patients with hepatic carcinoma. CASE PRESENTATION: We describe two cases of CLS after IL-11 administration in two males with thrombocytopenia. Case 1 was a 46-year-old man with recurrence of hepatic carcinoma who was treated with IL-11 (3 mg per day). After four days of therapy, hypotension and hypoproteinemia were detected. The chest X-ray and B ultrasound of the abdomen showed pleural effusion and ascites. IL-11 was then discontinued, fluid resuscitation was performed, and fresh frozen plasma and packed red blood cells were transfused into this patient. The patient had recovered after 19 days of treatment. Case 2 was a 66-year-old man who had undergone radiofrequency ablation (RFA) for hepatic carcinoma. He was treated with IL-11 (3 mg per day) for thrombocytopenia. After two days of therapy, this patient complained of dyspnea with bilateral edema of the hands. Laboratory values showed hypoproteinemia. IL-11 was stopped and human albumin was transfused at a rate of 10 g per day. On the 4th day, fluid resuscitation was performed. The patient had recovered after treatment for two weeks. CONCLUSIONS: The detection of IL-11-induced CLS supports the hypothesis that CLS could be a severe side effect of IL-11 treatment in some patients. These two case reports also demonstrate that patients with hepatic carcinoma who experience this rare form of CLS after treatment with IL-11 seem to respond to a therapeutic regimen that involves hydroxyethyl starch, albumin, and diuretic therapy. Liver cancer patients might be more susceptible to CLS because of poor liver function and hypersplenia. In addition, bleeding after RFA might be a further inducer of CLS. PMID- 21619579 TI - Prospective study of immunological factors in non-inflammatory bowel disease enterocutaneous fistulas. AB - BACKGROUND: Enterocutaneous fistulas (ECF) are debilitating and usually result following complex abdominal surgery. While there is an association with inflammatory bowel disease (IBD), a large number of fistulas occur after surgery not related to IBD. The consequences of ECF include short bowel syndrome and the need for long term parenteral nutrition.ECF can heal spontaneously and in the case of IBD can be cured by medical therapy in some instances. Those that do not resolve spontaneously have to be cured by surgery which is complex and associated with a high morbidity. It is not considered traditional treatment to use the same medical therapy as in IBD to cure ECF caused by other conditions.A small case series has reported three patients with persistent ECF not related to IBD to have healed following use of Infliximab which is the treatment commonly used for ECF caused by IBD. Infliximab acts by inhibiting the activity of the inflammatory cytokine TNF- alpha. It is not known if this cytokine is present in ECF tissue in the absence of IBD.The aim of this study is to demonstrate the presence of inflammatory markers in tissue surrounding non-IBD ECF and in particular to quantify the presence of the cytokine TNF- alpha. We hypothesise that TNF - alpha levels are raised in non-IBD ECF. METHODS/DESIGN: Tissue and serum from ECF of IBD and non-IBD patients will be prospectively collected at St. Mark's Hospital Intestinal Failure Unit. The control group will consist of patients undergoing colonoscopy for bowel cancer screening, with normal findings. Biopsies of the terminal ileum will be obtained from this group during colonoscopy. The fistula tract and serum cytokine profiles of interleukins (IL)-1a, IL-1b, IL-2, IL-4, IL 6, IL-8, IL-10, TNF- alpha, IFN-y, MCP-1, EGF and VEGF will be assessed. DISCUSSION: This study aims to assess the presence or absence of TNF- alpha expression in the ECF tissue in non-IBD origin. If our hypothesis is correct we would then be able to study the use of the TNF- alpha inhibitor Infliximab as a therapeutic option in the treatment of non-IBD ECF. Secondary aims include assessing the spectrum of inflammatory cytokines and markers present in tissue and serum of non-IBD ECF when compared with IBD ECF and normal controls. TRIAL REGISTRATION: ISRCTN44000447. PMID- 21619580 TI - Expert opinion on detecting and treating depression in palliative care: A Delphi study. AB - BACKGROUND: There is a dearth of data regarding the optimal method of detecting and treating depression in palliative care. This study applied the Delphi method to evaluate expert opinion on choice of screening tool, choice of antidepressant and choice of psychological therapy. The aim was to inform the development of best practice recommendations for the European Palliative Care Research Collaborative clinical practice guideline on managing depression in palliative care. METHODS: 18 members of an international, multi-professional expert group completed a structured questionnaire in two rounds, rating their agreement with proposed items on a scale from 0-10 and annotating with additional comments. The median and range were calculated to give a statistical average of the experts' ratings. RESULTS: There was contention regarding the benefits of screening, with 'routine informal asking' (median 8.5 (0-10)) rated more highly than formal screening tools such as the Hospital Anxiety and Depression Scale (median 7.0 (1 10). Mirtazapine (median 9 (7-10) and citalopram (median 9 (5-10) were the considered the best choice of antidepressant and cognitive behavioural therapy (median 9.0 (3-10) the best choice of psychological therapy. CONCLUSIONS: The range of expert ratings was broad, indicating discordance in the views of experts. Direct comparative data from randomised controlled trials are needed to strengthen the evidence-base and achieve clarity on how best to detect and treat depression in this setting. PMID- 21619581 TI - Differential gene expression of serine protease inhibitors in bovine ovarian follicle: possible involvement in follicular growth and atresia. AB - BACKGROUND: SERPINs (serine protease inhibitors) regulate proteases involving fibrinolysis, coagulation, inflammation, cell mobility, cellular differentiation and apoptosis. This study aimed to investigate differentially expressed genes of members of the SERPIN superfamily between healthy and atretic follicles using a combination of microarray and quantitative real-time PCR (QPCR) analysis. In addition, we further determined mRNA and protein localization of identified SERPINs in estradiol (E2)-active and E2-inactive follicles by in situ hybridization and immunohistochemistry. METHODS: We performed microarray analysis of healthy (10.7 +/- 0.7 mm) and atretic (7.8 +/- 0.2 mm) follicles using a custom-made bovine oligonucleotide microarray to screen differentially expressed genes encoding SERPIN superfamily members between groups. The expression profiles of six identified SERPIN genes were further confirmed by QPCR analysis. In addition, mRNA and protein localization of four SERPINs was investigated in E2 active and E2-inactive follicles using in situ hybridization and immunohistochemistry. RESULTS: We have identified 11 SERPIN genes expressed in healthy and atretic follicles by microarray analysis. QPCR analysis confirmed that mRNA expression of four SERPINs (SERPINA5, SERPINB6, SERPINE2 and SERPINF2) was greater in healthy than in atretic follicles, while two SERPINs (SERPINE1 and SERPING1) had greater expression in atretic than in healthy follicles. In situ hybridization showed that SERPINA5, SERPINB6 and SERPINF2 mRNA were localized in GCs of E2-active follicles and weakly expressed in GCs of E2-inactive follicles. SERPING1 mRNA was localized in both GCs and the theca layer (TL) of E2-inactive follicles and a weak hybridization signal was also detected in both GCs and TL of E2-active follicles. Immunohistochemistry showed that SERPINA5, SERPINB6 and SERPINF2 were detected in GCs of E2-active and E2-inactive follicles. SERPING1 protein was localized in both GCs and the TL of E2-active and E2-inactive follicles. CONCLUSIONS: Our results demonstrate a characteristic expression of SERPIN superfamily member genes in bovine healthy and atretic follicles. The cell type-and stage-specific expression of SERPINs may be associated with bovine follicular growth and atresia. PMID- 21619582 TI - Recent advances in nanoparticle formulation of oleanolic acid. AB - Oleanolic acid (OA) is a natural triterpenoid possessing anti-inflammatory, antitumor, antiviral, hepatoprotective and antihyperlipidemic effects. Research on the pharmacological activities and clinical applications of OA has made significant progress in the past decade, particularly in the areas such as isolation and purification, chemical modifications, pharmacological research, toxicity studies and clinical use of OA. However, due to its poor aqueous solubility, instability and low bioavailability, OA's clinical applications are still rather limited. Recently, nanoparticulate drug delivery as the biological dimension of nanotechnology has been developed, which may help generate useful formulations of OA for clinical applications. Nanoparticulate drug delivery system enhances the dissolution rate and bioavailability of OA, providing a feasible formulation method for clinical applications. PMID- 21619583 TI - Growth and development of Gnathostoma spinigerum (Nematoda: Gnathostomatidae) larvae in Mesocyclops aspericornis (Cyclopoida: Cyclopidae). AB - BACKGROUND: Gnathostoma spinigerum larva is pathogenic, causing gnathostomiasis in humans and certain animals, and is prevalent mainly in Asia. Growth and development of Gnathostoma spinigerum larvae in the cyclopoid copepod Mesocyclops aspericornis, the first intermediate host, were examined. RESULTS: When newly hatched, ensheathed second-stage larvae (L2) were ingested by M. aspericornis, they immediately appeared exsheathed in the stomach of M. aspericornis. They then penetrated the stomach wall and entered the body cavity, where they immediately metamorphosed to a stunted form with the body length/width ratio equal to the early third-stage larvae (EL3) up to 2 h after being ingested. During metamorphosis, the anterior spine-like structure of L2 transformed into unequal transparent lips. The larvae moulted into EL3 in the body cavity of the copepod at around day 5-7 post-infection. Minute cuticular striations were seen on the whole body, with prominent single-pointed spines on the anterior part of the body. The head bulb had four rows of hooklets and two lateral trilobed lips. The size of EL3 in copepods continuously increased towards day 12 and showed a negative correlation to their density per copepod (R = -0.881, P < 0.05 for body length, and R = -0.906, P < 0.05 for body width). CONCLUSIONS: The results revealed for the first time that M. aspericornis, one of the most abundant freshwater copepods in Thailand, is a suitable first intermediate host for G. spinigerum. High susceptibility of M. aspericornis suggests its importance for the maintenance of the life cycle of G. spinigerum in Thailand. PMID- 21619584 TI - Intra-ocular melanoma metastatic to an axillary lymph node: a case report. AB - BACKGROUND: Unusual metastatic presentation of intra-ocular melanoma. STUDY DESIGN: Case report. DISCUSSION: Extra-regional lymphatic spread of intra-ocular melanoma has not been reported previously in the literature. The usual pattern of metastasis for intra-ocular melanoma is hematogenous. There are few reports of regional spread to the maxillofacial bones. We report an interesting case of a 51 year old female with prior history of right eye melanoma, now presenting with metastasis to the left axilla, which is an extra-regional nodal basin. CONCLUSION: In female patients presenting with an isolated axillary mass, with a negative breast work up and known prior history of melanoma, the differential diagnosis should include possible metastatic melanoma. Core biopsy will confirm the diagnosis and tailor subsequent management. PMID- 21619585 TI - The current shortage and future surplus of doctors: a projection of the future growth of the Japanese medical workforce. AB - BACKGROUND: Starting in the late 1980s, the Japanese government decreased the number of students accepted into medical school each year in order to reduce healthcare spending. The result of this policy is a serious shortage of doctors in Japan today, which has become a social problem in recent years. In an attempt to solve this problem, the Japanese government decided in 2007 to increase the medical student quota from 7625 to 8848. Furthermore, the Democratic Party of Japan (DPJ), Japan's ruling party after the 2009 election, promised in their manifesto to increase the medical student quota to 1.5 times what it was in 2007, in order to raise the number of medical doctors to more than 3.0 per 1000 persons. It should be noted, however, that this rapid increase in the medical student quota may bring about a serious doctor surplus in the future, especially because the population of Japan is decreasing.The purpose of this research is to project the future growth of the Japanese medical doctor workforce from 2008 to 2050 and to forecast whether the proposed additional increase in the student quota will cause a doctor surplus. METHODS: Simulation modeling of the Japanese medical workforce. RESULTS: Even if the additional increase in the medical student quota promised by the DPJ fails, the number of practitioners is projected to increase from 286 699 (2.25 per 1000 persons) in 2008 to 365 533 (over the national numerical goal of 3.0 per 1000) in 2024. The number of practitioners per 1000 persons is projected to further increase to 3.10 in 2025, to 3.71 in 2035, and to 4.69 in 2050. If the additional increase in the medical student quota promised by the DPJ is realized, the total workforce is projected to rise to 392 331 (3.29 per 1000 persons) in 2025, 464 296 (4.20 per 1,000 persons) in 2035, and 545 230 (5.73 per 1000 persons) in 2050. CONCLUSIONS: The plan to increase the medical student quota will bring about a serious doctor surplus in the long run. PMID- 21619586 TI - Systemic chemotherapy for treatment of advanced small bowel adenocarcinoma with prognostic factor analysis: retrospective study. AB - BACKGROUND: We sought to evaluate prognostic factors affecting overall survival (OS), and to investigate the role of palliative chemotherapy using propensity score-based weighting, in patients with advanced small bowel adenocarcinoma (SBA). METHODS: Data from a total of 91 patients diagnosed with advanced SBA at the Asan Medical Center between January 1989 and December 2009 were retrospectively analyzed. Patients were split into two groups, those who did and did not receive palliative chemotherapy. RESULTS: Overall, 81 patients (89.0%) died, at a median survival time of 6.6 months (95% confidence interval [CI], 5.5 7.5 months). The 40 patients receiving chemotherapy showed overall response and disease control rates of 11.1% and 37.0%, respectively, with OS and progression free survival (PFS) of 11.8 months (95% CI, 4.6 - 19.0 months) and 5.7 months (95% CI, 3.5 - 8.0 months), respectively. The 41 patients who did not receive chemotherapy had an OS of 4.1 months (95% CI, 3.1 - 5.1 months) and a PFS of 1.3 months (95% CI, 0.8 - 1.7 months). Multivariate analysis showed that lack of tumor resection, non-prescription of chemotherapy, liver metastasis, and intra abdominal lymph node metastasis, were all independently associated with poor survival outcomes. After inverse probability of treatment weighting (IPTW) adjustment, the group that did not receive chemotherapy was at a significantly higher risk of mortality (HR 3.44, 95% CI 2.03 - 5.83, p < 0.001) than were patients receiving chemotherapy. CONCLUSION: Palliative chemotherapy may improve survival outcomes in patients with advanced SBA. PMID- 21619587 TI - PKC isoforms interact with and phosphorylate DNMT1. AB - BACKGROUND: DNA methyltransferase 1 (DNMT1) has been shown to be phosphorylated on multiple serine and threonine residues, based on cell type and physiological conditions. Although recent studies have suggested that protein kinase C (PKC) may be involved, the individual contribution of PKC isoforms in their ability to phosphorylate DNMT1 remains unknown. The PKC family consists of at least 12 isoforms that possess distinct differences in structure, substrate requirement, expression and localization. RESULTS: Here we show that PKCalpha, betaI, betaII, delta, gamma, eta, zeta and MU preferentially phosphorylate the N-terminal domain of human DNMT1. No such phosphorylation of DNMT1 was observed with PKCepsilon. Using PKCzeta as a prototype model, we also found that PKC physically interacts with and phosphorylates DNMT1. In vitro phosphorylation assays conducted with recombinant fragments of DNMT1 showed that PKCzeta preferentially phosphorylated the N-terminal region of DNMT1. The interaction of PKCzeta with DNMT1 was confirmed by GST pull-down and co-immunoprecipitation experiments. Co localization experiments by fluorescent microscopy further showed that endogenous PKCzeta and DNMT1 were present in the same molecular complex. Endogenous PKCzeta activity was also detected when DNMT1 was immunoprecipitated from HEK-293 cells. Overexpression of both PKCzeta and DNMT1 in HEK-293 cells, but not of either alone, reduced the methylation status of genes distributed across the genome. Moreover, in vitro phosphorylation of DNMT1 by PKCzeta reduced its methytransferase activity. CONCLUSIONS: Our results indicate that phosphorylation of human DNMT1 by PKC is isoform-specific and provides the first evidence of cooperation between PKCzeta and DNMT1 in the control of the DNA methylation patterns of the genome. PMID- 21619588 TI - Predictive performance of the visceral adiposity index for a visceral adiposity related risk: type 2 diabetes. AB - BACKGROUND: Visceral adiposity index (VAI) has recently been developed based on waist circumference, body mass index (BMI), triglycerides (TGs), and high-density lipoprotein cholesterol (HDL-C). We examined predictive performances for incident diabetes of the VAI per se and as compared to the metabolic syndrome (MetS) and waist-to-height-ratio (WHtR). METHODS: Participants free of diabetes at baseline with at least one follow-up examination (5,964) were included for the current study. Weibull regression models were developed for interval-censored survival data. Absolute and relative integrated discriminatory improvement index (IDI) and cut-point-based and cut-point-free net reclassification improvement index (NRI) were used as measures of predictive ability for incident diabetes added by VAI, as compared to the MetS and WHtR. RESULTS: The annual incidence rate of diabetes was 0.85 per 1000 person. Mean VAI was 3.06 (95%CIs 2.99-3.13). Diabetes risk factors levels increased in stepwise fashion across VAI quintiles. Risk gradient between the highest and lowest quintile of VAI was 4.5 (95%CIs 3.0-6.9). VAI significantly improved predictive ability of the MetS. The relative IDI and cut point free NRI for predictive ability added to MetS by VAI were 30.3% (95%CIs 18.8-41.8%) and 30.7% (95%CIs 20.8-40.7%), respectively. WHtR, outperformed VAI with cut-point-free NRI of 24.6% (95%CIs 14.1-35.2%). CONCLUSIONS: In conclusion, although VAI could be a prognostic tool for incident diabetes events, gathering information on its components (WC, BMI, TGs, and HDL-C) is unlikely to improve the prediction ability beyond what could be achieved by the simply assessable and commonly available information on WHtR. PMID- 21619589 TI - Single-cell time-lapse analysis of depletion of the universally conserved essential protein YgjD. AB - BACKGROUND: The essential Escherichia coli gene ygjD belongs to a universally conserved group of genes whose function has been the focus of a number of recent studies. Here, we put ygjD under control of an inducible promoter, and used time lapse microscopy and single cell analysis to investigate the phenotypic consequences of the depletion of YgjD protein from growing cells. RESULTS: We show that loss of YgjD leads to a marked decrease in cell size and termination of cell division. The transition towards smaller size occurs in a controlled manner: cell elongation and cell division remain coupled, but cell size at division decreases. We also find evidence that depletion of YgjD leads to the synthesis of the intracellular signaling molecule (p)ppGpp, inducing a cellular reaction resembling the stringent response. Concomitant deletion of the relA and spoT genes - leading to a strain that is uncapable of synthesizing (p)ppGpp - abrogates the decrease in cell size, but does not prevent termination of cell division upon YgjD depletion. CONCLUSIONS: Depletion of YgjD protein from growing cells leads to a decrease in cell size that is contingent on (p)ppGpp, and to a termination of cell division. The combination of single-cell timelapse microscopy and statistical analysis can give detailed insights into the phenotypic consequences of the loss of essential genes, and can thus serve as a new tool to study the function of essential genes. PMID- 21619590 TI - Real-time visualization of heterotrimeric G protein Gq activation in living cells. AB - BACKGROUND: Gq is a heterotrimeric G protein that plays an important role in numerous physiological processes. To delineate the molecular mechanisms and kinetics of signalling through this protein, its activation should be measurable in single living cells. Recently, fluorescence resonance energy transfer (FRET) sensors have been developed for this purpose. RESULTS: In this paper, we describe the development of an improved FRET-based Gq activity sensor that consists of a yellow fluorescent protein (YFP)-tagged Ggamma2 subunit and a Galphaq subunit with an inserted monomeric Turquoise (mTurquoise), the best cyan fluorescent protein variant currently available. This sensor enabled us to determine, for the first time, the kon (2/s) of Gq activation. In addition, we found that the guanine nucleotide exchange factor p63RhoGEF has a profound effect on the number of Gq proteins that become active upon stimulation of endogenous histamine H1 receptors. The sensor was also used to measure ligand-independent activation of the histamine H1 receptor (H1R) upon addition of a hypotonic stimulus. CONCLUSIONS: Our observations reveal that the application of a truncated mTurquoise as donor and a YFP-tagged Ggamma2 as acceptor in FRET-based Gq activity sensors substantially improves their dynamic range. This optimization enables the real-time single cell quantification of Gq signalling dynamics, the influence of accessory proteins and allows future drug screening applications by virtue of its sensitivity. PMID- 21619591 TI - Double chambered right ventricle with severe calcification of the tricuspid valve in an elderly woman: a case report. AB - INTRODUCTION: Double chambered right ventricle is a rare congenital cardiac anomaly in which the right ventricle is divided into two chambers by an anomalous muscle bundle. The diagnosis of this disorder is difficult in adults. Calcification of the tricuspid valve is extremely rare, and very few cases have been reported. Most cases of tricuspid valve calcification had a congenital disorder with high pressure in the right ventricle. CASE PRESENTATION: We report a rare case of a 71-year-old Japanese woman who presented with chest discomfort, and was found to have a double chambered right ventricle with severe calcification of the tricuspid valve. This abnormality was found by echocardiography, and the diagnosis was confirmed by multislice cardiac computerized tomography, cardiac magnetic resonance imaging, and cardiac catheterization. Our patient rejected surgical repair, and medical therapy with carvedilol was effective to reduce her symptoms. CONCLUSION: Calcification of the tricuspid valve is extremely rare, and considered to be due to high pressure in the right ventricle. To the best of our knowledge, there are no other reported cases of this combination of double chambered right ventricle and calcification of the tricuspid valve. PMID- 21619592 TI - Early diagnosis of peripheral nervous system involvement in Fabry disease and treatment of neuropathic pain: the report of an expert panel. AB - BACKGROUND: Fabry disease is an inherited metabolic disorder characterized by progressive lysosomal accumulation of lipids in a variety of cell types, including neural cells. Small, unmyelinated nerve fibers are particularly affected and small fiber peripheral neuropathy often clinically manifests at young age. Peripheral pain can be chronic and/or occur as provoked attacks of excruciating pain. Manifestations of dysfunction of small autonomic fibers may include, among others, impaired sweating, gastrointestinal dysmotility, and abnormal pain perception. Patients with Fabry disease often remain undiagnosed until severe complications involving the kidney, heart, peripheral nerves and/or brain have arisen. METHODS: An international expert panel convened with the goal to provide guidance to clinicians who may encounter unrecognized patients with Fabry disease on how to diagnose these patients early using simple diagnostic tests. A further aim was to offer recommendations to control neuropathic pain. RESULTS: We describe the neuropathy in Fabry disease, focusing on peripheral small fiber dysfunction - the hallmark of early neurologic involvement in this disorder. The clinical course of peripheral pain is summarized, and the importance of medical history-taking, including family history, is highlighted. A thorough physical examination (e.g., angiokeratoma, corneal opacities) and simple non-invasive sensory perception tests could provide clues to the diagnosis of Fabry disease. Reported early clinical benefits of enzyme replacement therapy include reduction of neuropathic pain, and adequate management of residual pain to a tolerable and functional level can substantially improve the quality of life for patients. CONCLUSIONS: Our recommendations can assist in diagnosing Fabry small fiber neuropathy early, and offer clinicians guidance in controlling peripheral pain. This is particularly important since management of pain in young patients with Fabry disease appears to be inadequate. PMID- 21619593 TI - Left ventricular geometric patterns and adaptations to hemodynamics are similar in elderly men and women. AB - BACKGROUND: Common conditions such as obesity and hypertension result in hemodynamic alterations that will induce remodeling of the left ventricle (LV). However, differences between the genders in the relationship of hemodynamics to LV geometry are not well known.The present study aims to investigate differences between the genders in this respect, in a sample of elderly persons. METHODS: Echocardiography and Doppler was performed in a population-based sample aged 70 - The Prospective Investigation of the Vasculature in Uppsala Seniors (PIVUS) study (n = 922).Hemodynamic patterns obtained by echocardiography and Doppler were evaluated in relation to four LV geometric groups (normal, concentric remodeling, eccentric hypertrophy and concentric hypertrophy). RESULTS: No significant difference between the genders was observed regarding the prevalence of the LV geometric groups.Mean values of most evaluated echocardiography and Doppler variables differed between men and women, such as LA, IVS, LVEDD and IVRT, but the relationship of hemodynamic variables to LV geometric groups did not differ between the genders. CONCLUSIONS: Although mean values of many echocardiographic variables differed between men and women, the LV geometric adaptations to a given hemodynamic load appear similar in both genders. PMID- 21619594 TI - Variations in Mre11/Rad50/Nbs1 status and DNA damage-induced S-phase arrest in the cell lines of the NCI60 panel. AB - BACKGROUND: The Mre11/Rad50/Nbs1 (MRN) complex is a regulator of cell cycle checkpoints and DNA repair. Defects in MRN can lead to defective S-phase arrest when cells are damaged. Such defects may elicit sensitivity to selected drugs providing a chemical synthetic lethal interaction that could be used to target therapy to tumors with these defects. The goal of this study was to identify these defects in the NCI60 panel of cell lines and identify compounds that might elicit selective cytotoxicity. METHODS: We screened the NCI60 panel in search of cell lines that express low levels of MRN proteins, or that fail to arrest in S phase in response to the topisomerase I inhibitor SN38. The NCI COMPARE program was used to discover compounds that preferentially target cells with these phenotypes. RESULTS: HCT116 cells were initially identified as defective in MRN and S phase arrest. Transfection with Mre11 also elevated Rad50 and Nbs1, and rescued the defective S-phase arrest. Cells of the NCI60 panel exhibited a large range of protein expression but a strong correlation existed between Mre11, Rad50 and Nbs1 consistent with complex formation determining protein stability. Mre11 mRNA correlated best with protein level suggesting it was the primary determinant of the overall level of the complex. Three other cell lines failed to arrest in response to SN38, two of which also had low MRN. However, other cell lines with low MRN still arrested suggesting low MRN does not predict an inability to arrest. Many compounds, including a family of benzothiazoles, correlated with the failure to arrest in S phase. The activity of benzothiazoles has been attributed to metabolic activation and DNA alkylation, but we note several cell lines in which sensitivity does not correlate with metabolism. We propose that the checkpoint defect imposes an additional mechanism of sensitivity on cells. CONCLUSIONS: We have identified cells with possible defects in the MRN complex and S phase arrest, and a series of compounds that may preferentially target S phase-defective cells. We discuss limitations of the COMPARE program when attempting to identify compounds that selectively inhibit only a few cell lines. PMID- 21619595 TI - Unravelling the effects of age, period and cohort on metabolic syndrome components in a Taiwanese population using partial least squares regression. AB - BACKGROUND: We investigate whether the changing environment caused by rapid economic growth yielded differential effects for successive Taiwanese generations on 8 components of metabolic syndrome (MetS): body mass index (BMI), systolic blood pressure (SBP), diastolic blood pressure (DBP), fasting plasma glucose (FPG), triglycerides (TG), high-density lipoprotein (HDL), Low-density lipoproteins (LDL) and uric acid (UA). METHODS: To assess the impact of age, birth year and year of examination on MetS components, we used partial least squares regression to analyze data collected by Mei-Jaw clinics in Taiwan in years 1996 and 2006. Confounders, such as the number of years in formal education, alcohol intake, smoking history status, and betel-nut chewing were adjusted for. RESULTS: As the age of individuals increased, the values of components generally increased except for UA. Men born after 1970 had lower FPG, lower BMI, lower DBP, lower TG, Lower LDL and greater HDL; women born after 1970 had lower BMI, lower DBP, lower TG, Lower LDL and greater HDL and UA. There is a similar pattern between the trend in levels of metabolic syndrome components against birth year of birth and economic growth in Taiwan. CONCLUSIONS: We found cohort effects in some MetS components, suggesting associations between the changing environment and health outcomes in later life. This ecological association is worthy of further investigation. PMID- 21619596 TI - Integrating telecare for chronic disease management in the community: what needs to be done? AB - BACKGROUND: Telecare could greatly facilitate chronic disease management in the community, but despite government promotion and positive demonstrations its implementation has been limited. This study aimed to identify factors inhibiting the implementation and integration of telecare systems for chronic disease management in the community. METHODS: Large scale comparative study employing qualitative data collection techniques: semi-structured interviews with key informants, task-groups, and workshops; framework analysis of qualitative data informed by Normalization Process Theory. Drawn from telecare services in community and domestic settings in England and Scotland, 221 participants were included, consisting of health professionals and managers; patients and carers; social care professionals and managers; and service suppliers and manufacturers. RESULTS: Key barriers to telecare integration were uncertainties about coherent and sustainable service and business models; lack of coordination across social and primary care boundaries, lack of financial or other incentives to include telecare within primary care services; a lack of a sense of continuity with previous service provision and self-care work undertaken by patients; and general uncertainty about the adequacy of telecare systems. These problems led to poor integration of policy and practice. CONCLUSION: Telecare services may offer a cost effective and safe form of care for some people living with chronic illness. Slow and uneven implementation and integration do not stem from problems of adoption. They result from incomplete understanding of the role of telecare systems and subsequent adaption and embeddedness to context, and uncertainties about the best way to develop, coordinate, and sustain services that assist with chronic disease management. Interventions are therefore needed that (i) reduce uncertainty about the ownership of implementation processes and that lock together health and social care agencies; and (ii) ensure user centred rather than biomedical/service-centred models of care. PMID- 21619597 TI - Indole and 3-indolylacetonitrile inhibit spore maturation in Paenibacillus alvei. AB - BACKGROUND: Bacteria use diverse signaling molecules to ensure the survival of the species in environmental niches. A variety of both gram-positive and gram negative bacteria produce large quantities of indole that functions as an intercellular signal controlling diverse aspects of bacterial physiology. RESULTS: In this study, we sought a novel role of indole in a gram-positive bacteria Paenibacillus alvei that can produce extracellular indole at a concentration of up to 300 MUM in the stationary phase in Luria-Bertani medium. Unlike previous studies, our data show that the production of indole in P. alvei is strictly controlled by catabolite repression since the addition of glucose and glycerol completely turns off the indole production. The addition of exogenous indole markedly inhibits the heat resistance of P. alvei without affecting cell growth. Observation of cell morphology with electron microscopy shows that indole inhibits the development of spore coats and cortex in P. alvei. As a result of the immature spore formation of P. alvei, indole also decreases P. alvei survival when exposed to antibiotics, low pH, and ethanol. Additionally, indole derivatives also influence the heat resistance; for example, a plant auxin, 3 indolylacetonitrile dramatically (2900-fold) decreased the heat resistance of P. alvei, while another auxin 3-indoleacetic acid had a less significant influence on the heat resistance of P. alvei. CONCLUSIONS: Together, our results demonstrate that indole and plant auxin 3-indolylacetonitrile inhibit spore maturation of P. alvei and that 3-indolylacetonitrile presents an opportunity for the control of heat and antimicrobial resistant spores of gram-positive bacteria. PMID- 21619598 TI - Yellow fever virus envelope protein expressed in insect cells is capable of syncytium formation in lepidopteran cells and could be used for immunodetection of YFV in human sera. AB - BACKGROUND: Yellow fever is an haemorrhagic disease caused by a virus that belongs to the genus Flavivirus (Flaviviridae family) and is transmitted by mosquitoes. Among the viral proteins, the envelope protein (E) is the most studied one, due to its high antigenic potencial. Baculovirus are one of the most popular and efficient eukaryotic expression system. In this study a recombinant baculovirus (vSynYFE) containing the envelope gene (env) of the 17D vaccine strain of yellow fever virus was constructed and the recombinant protein antigenicity was tested. RESULTS: Insect cells infected with vSynYFE showed syncytium formation, which is a cytopathic effect characteristic of flavivirus infection and expressed a polypeptide of around 54 kDa, which corresponds to the expected size of the recombinant E protein. Furthermore, the recombinant E protein expression was also confirmed by fluorescence microscopy of vSynYFE infected insect cells. Total vSynYFE-infected insect extracts used as antigens detected the presence of antibodies for yellow fever virus in human sera derived from yellow fever-infected patients in an immunoassay and did not cross react with sera from dengue virus-infected patients. CONCLUSIONS: The E protein expressed by the recombinant baculovirus in insect cells is antigenically similar to the wild protein and it may be useful for different medical applications, from improved diagnosis of the disease to source of antigens for the development of a subunit vaccine. PMID- 21619599 TI - Claudins in lung diseases. AB - Tight junctions are the most apically localized part of the epithelial junctional complex. They regulate the permeability and polarity of cell layers and create compartments in cell membranes. Claudins are structural molecules of tight junctions. There are 27 claudins known, and expression of different claudins is responsible for changes in the electrolyte and solute permeability in cells layers. Studies have shown that claudins and tight junctions also protect multicellular organisms from infections and that some infectious agents may use claudins as targets to invade and weaken the host's defense. In neoplastic diseases, claudin expression may be up- or downregulated. Since their expression is associated with specific tumor types or with specific locations of tumors to a certain degree, they can, in a restricted sense, also be used as tumor markers. However, the regulation of claudin expression is complex involving growth factors and integrins, protein kinases, proto-oncogens and transcription factors. In this review, the significance of claudins is discussed in lung disease and development. PMID- 21619600 TI - A physical map for the Amborella trichopoda genome sheds light on the evolution of angiosperm genome structure. AB - BACKGROUND: Recent phylogenetic analyses have identified Amborella trichopoda, an understory tree species endemic to the forests of New Caledonia, as sister to a clade including all other known flowering plant species. The Amborella genome is a unique reference for understanding the evolution of angiosperm genomes because it can serve as an outgroup to root comparative analyses. A physical map, BAC end sequences and sample shotgun sequences provide a first view of the 870 Mbp Amborella genome. RESULTS: Analysis of Amborella BAC ends sequenced from each contig suggests that the density of long terminal repeat retrotransposons is negatively correlated with that of protein coding genes. Syntenic, presumably ancestral, gene blocks were identified in comparisons of the Amborella BAC contigs and the sequenced Arabidopsis thaliana, Populus trichocarpa, Vitis vinifera and Oryza sativa genomes. Parsimony mapping of the loss of synteny corroborates previous analyses suggesting that the rate of structural change has been more rapid on lineages leading to Arabidopsis and Oryza compared with lineages leading to Populus and Vitis. The gamma paleohexiploidy event identified in the Arabidopsis, Populus and Vitis genomes is shown to have occurred after the divergence of all other known angiosperms from the lineage leading to Amborella. CONCLUSIONS: When placed in the context of a physical map, BAC end sequences representing just 5.4% of the Amborella genome have facilitated reconstruction of gene blocks that existed in the last common ancestor of all flowering plants. The Amborella genome is an invaluable reference for inferences concerning the ancestral angiosperm and subsequent genome evolution. PMID- 21619601 TI - Path finding methods accounting for stoichiometry in metabolic networks. AB - Graph-based methods have been widely used for the analysis of biological networks. Their application to metabolic networks has been much discussed, in particular noting that an important weakness in such methods is that reaction stoichiometry is neglected. In this study, we show that reaction stoichiometry can be incorporated into path-finding approaches via mixed-integer linear programming. This major advance at the modeling level results in improved prediction of topological and functional properties in metabolic networks. PMID- 21619602 TI - New Topoisomerase I mutations are associated with resistance to camptothecin. AB - BACKGROUND: Topoisomerase I (TOP1) is a nuclear enzyme that catalyzes the relaxation of supercoiled DNA during DNA replication and transcription. TOP1 is the molecular target of camptothecin and related drugs such as irinotecan and SN38 (irinotecan's active metabolite). Irinotecan is widely used as an anti cancer agent in the treatment of metastatic colon cancer. However, its efficacy is often limited by the development of resistance. METHODS: We previously established several SN38 resistant HCT116-derived clones to study the mechanisms underlying resistance to SN38. Here, we investigated whether resistance to SN38 in these cell lines could be linked to the presence of TOP1 mutations and changes in its expression and activity. Functional analyses were performed on these cell lines challenged with SN38 and we specifically monitored the double strands breaks with gammaH2AX staining and replication activity with molecular combing. RESULTS: In SN38 resistant HCT116 clones we identified three new TOP1 mutations, which are located in the core subdomain III (p.R621H and p.L617I) and in the linker domain (p.E710G) and are packed together at the interface between these two domains. The presence of these TOP1 mutations in SN38 resistant HCT116 cells did not modify TOP1 expression or intrinsic activity. Conversely, following challenge with SN38, we observed a decrease of TOP1-DNA cleavage complexes and a reduction in double-stranded break formation). In addition, we showed that SN38 resistant HCT116 cells present a strong decrease in the SN38-dependent asymmetry of replication forks that is characteristic of SN38 sensitive HCT116 cells. CONCLUSIONS: These results indicate that the TOP1 mutations are involved in the development of SN38 resistance. We hypothesize that p.L617, p.R621 and p.E710 TOP1 residues are important for the functionality of the linker and that mutation of one of these residues is sufficient to alter or modulate its flexibility. Consequently, linker fluctuations could have an impact on SN38 binding by reducing the enzyme affinity for the drug. PMID- 21619603 TI - Exploring subdomain variation in biomedical language. AB - BACKGROUND: Applications of Natural Language Processing (NLP) technology to biomedical texts have generated significant interest in recent years. In this paper we identify and investigate the phenomenon of linguistic subdomain variation within the biomedical domain, i.e., the extent to which different subject areas of biomedicine are characterised by different linguistic behaviour. While variation at a coarser domain level such as between newswire and biomedical text is well-studied and known to affect the portability of NLP systems, we are the first to conduct an extensive investigation into more fine-grained levels of variation. RESULTS: Using the large OpenPMC text corpus, which spans the many subdomains of biomedicine, we investigate variation across a number of lexical, syntactic, semantic and discourse-related dimensions. These dimensions are chosen for their relevance to the performance of NLP systems. We use clustering techniques to analyse commonalities and distinctions among the subdomains. CONCLUSIONS: We find that while patterns of inter-subdomain variation differ somewhat from one feature set to another, robust clusters can be identified that correspond to intuitive distinctions such as that between clinical and laboratory subjects. In particular, subdomains relating to genetics and molecular biology, which are the most common sources of material for training and evaluating biomedical NLP tools, are not representative of all biomedical subdomains. We conclude that an awareness of subdomain variation is important when considering the practical use of language processing applications by biomedical researchers. PMID- 21619604 TI - Potential pitfalls of modelling ribosomal RNA data in phylogenetic tree reconstruction: evidence from case studies in the Metazoa. AB - BACKGROUND: Failure to account for covariation patterns in helical regions of ribosomal RNA (rRNA) genes has the potential to misdirect the estimation of the phylogenetic signal of the data. Furthermore, the extremes of length variation among taxa, combined with regional substitution rate variation can mislead the alignment of rRNA sequences and thus distort subsequent tree reconstructions. However, recent developments in phylogenetic methodology now allow a comprehensive integration of secondary structures in alignment and tree reconstruction analyses based on rRNA sequences, which has been shown to correct some of these problems. Here, we explore the potentials of RNA substitution models and the interactions of specific model setups with the inherent pattern of covariation in rRNA stems and substitution rate variation among loop regions. RESULTS: We found an explicit impact of RNA substitution models on tree reconstruction analyses. The application of specific RNA models in tree reconstructions is hampered by interaction between the appropriate modelling of covarying sites in stem regions, and excessive homoplasy in some loop regions. RNA models often failed to recover reasonable trees when single-stranded regions are excessively homoplastic, because these regions contribute a greater proportion of the data when covarying sites are essentially downweighted. In this context, the RNA6A model outperformed all other models, including the more parametrized RNA7 and RNA16 models. CONCLUSIONS: Our results depict a trade-off between increased accuracy in estimation of interdependencies in helical regions with the risk of magnifying positions lacking phylogenetic signal. We can therefore conclude that caution is warranted when applying rRNA covariation models, and suggest that loop regions be independently screened for phylogenetic signal, and eliminated when they are indistinguishable from random noise. In addition to covariation and homoplasy, other factors, like non-stationarity of substitution rates and base compositional heterogeneity, can disrupt the signal of ribosomal RNA data. All these factors dictate sophisticated estimation of evolutionary pattern in rRNA data, just as other molecular data require similarly complicated (but different) corrections. PMID- 21619605 TI - Antiprogestin mifepristone inhibits the growth of cancer cells of reproductive and non-reproductive origin regardless of progesterone receptor expression. AB - BACKGROUND: Mifepristone (MF) has been largely used in reproductive medicine due to its capacity to modulate the progesterone receptor (PR). The study of MF has been expanded to the field of oncology; yet it remains unclear whether the expression of PR is required for MF to act as an anti-cancer agent. Our laboratory has shown that MF is a potent inhibitor of ovarian cancer cell growth. In this study we questioned whether the growth inhibitory properties of MF observed in ovarian cancer cells would translate to other cancers of reproductive and non-reproductive origin and, importantly, whether its efficacy is related to the expression of cognate PR. METHODS: Dose-response experiments were conducted with cancer cell lines of the nervous system, breast, prostate, ovary, and bone. Cultures were exposed to vehicle or increasing concentrations of MF for 72 h and analysed for cell number and cell cycle traverse, and hypodiploid DNA content characteristic of apoptotic cell death. For all cell lines, expression of steroid hormone receptors upon treatment with vehicle or cytostatic doses of MF for 24 h was studied by Western blot, whereas the activity of the G1/S regulatory protein Cdk2 in both treatment groups was monitored in vitro by the capacity of Cdk2 to phosphorylate histone H1. RESULTS: MF growth inhibited all cancer cell lines regardless of tissue of origin and hormone responsiveness, and reduced the activity of Cdk2. Cancer cells in which MF induced G1 growth arrest were less susceptible to lethality in the presence of high concentrations of MF, when compared to cancer cells that did not accumulate in G1. While all cancer cell lines were growth inhibited by MF, only the breast cancer MCF-7 cells expressed cognate PR. CONCLUSIONS: Antiprogestin MF inhibits the growth of different cancer cell lines with a cytostatic effect at lower concentrations in association with a decline in the activity of the cell cycle regulatory protein Cdk2, and apoptotic lethality at higher doses in association with increased hypodiploid DNA content. Contrary to common opinion, growth inhibition of cancer cells by antiprogestin MF is not dependent upon expression of classical, nuclear PR. PMID- 21619606 TI - Health-related quality of life in French adolescents and adults: norms for the DUKE Health Profile. AB - BACKGROUND: The continual monitoring of population health-related quality of life (HRQoL) with validated instruments helps public health agencies assess, protect, and promote population health. This study aimed to determine norms for the French adolescent and adult general population for the Duke Health Profile (DUKE) questionnaire in a large representative community sample. METHODS: We randomly selected 17,733 French people aged 12 to 75 years old in 2 steps, by households and individuals, from the National Health Barometer 2005, a periodic population study by the French National Institute for Prevention and Health Education. Quality of life and other data were collected by computer-assisted telephone interview. RESULTS: Normative data for the French population were analyzed by age, gender and self-reported chronic disease. Globally, function scores (best HRQoL=100) for physical, mental, social, and general health, as well as perceived health and self-esteem, were 72.3 (SEM 0.2), 74.6 (0.2), 66.8 (0.1), 71.3 (0.1), 71.3 (0.3), 76.5 (0.1), respectively. Dysfunction scores (worst HRQoL=100) for anxiety, depression, pain and disability domains were 30.9 (0.1), 27.6 (0.2), 34.3 (0.3), 3.1 (0.1), respectively. CONCLUSION: The French norms for adolescents and adults for the DUKE could be used as a reference for other studies assessing HRQoL, for specific illnesses, in France and for international comparisons. PMID- 21619607 TI - The functional status and well being of people with myalgic encephalomyelitis/chronic fatigue syndrome and their carers. AB - BACKGROUND: Diagnosis of myalgic encephalomyelitis/chronic fatigue syndrome or ME/CFS is largely based on clinical history, and exclusion of identifiable causes of chronic fatigue. Characterization of cases and the impact of interventions have been limited due to clinical heterogeneity and a lack of reliable biomarkers for diagnosis and outcome measures. People with ME/CFS (PWME) often report high levels of disability, which are difficult to measure objectively. The well being of family members and those who care for PWME are also likely to be affected. This study aimed to investigate the functional status and well being of PWME and their lay carers, and to compare them with people with other chronic conditions. METHODS: We used a cross sectional design to study 170 people aged between 18 and 64 years with well characterized ME/CFS, and 44 carers, using SF-36 v2TM. Mean physical and mental domains scores (scales and component summaries) were calculated and compared internally and externally with reference standards for the general population and for population groups with 10 chronic diseases. RESULTS: SF-36 scores in PWME were significantly reduced, especially within the physical domain (mean norm-based Physical Component Summary (PCS) score = 26.8), but also within the mental domain (mean norm-based score for Mental Component Summary (MCS) = 34.1). The lowest and highest scale scores were for "Role Physical" (mean = 25.4) and "Mental Health" (mean = 36.7) respectively. All scores were in general lower than those for the general population and diseased specific norms for other diseases. Carers of those with ME/CFS tended to have low scores in relation to population norms, particularly within the mental domain (mean = 45.4). CONCLUSIONS: ME/CFS is disabling and has a greater impact on functional status and well being than other chronic diseases such as cancer. The emotional burden of ME/CFS is felt by lay carers as well as by people with ME/CFS. We suggest the use of generic instruments such as SF-36, in combination of other objective outcome measurements, to describe patients and assess treatments. PMID- 21619608 TI - Psychiatry out-of-hours: a focus group study of GPs' experiences in Norwegian casualty clinics. AB - BACKGROUND: For Norwegian general practitioners (GPs), acute treatment of mental illness and substance abuse are among the most commonly experienced emergency situations in out-of-hours primary healthcare. The largest share of acute referrals to emergency psychiatric wards occurs out-of-hours, and out-of-hours services are responsible for a disproportionately high share of compulsory referrals. Concerns exist regarding the quality of mental healthcare provided in the out-of-hours setting. The aim of this study was to explore which challenges GPs experience when providing emergency care out-of-hours to patients presenting problems related to mental illness or substance abuse. METHODS: We conducted a qualitative study based on two individual interviews and six focus groups with purposively sampled GPs (totally 45 participants). The interviews were analysed successively in an editing style, using a thematic approach based on methodological descriptions by Charmaz and Malterud. RESULTS: Safety and uncertainty were the dominating themes in the discussions. The threat to personal safety due to unpredictable patient behaviour was a central concern, and present security precautions in the out-of-hours services were questioned. The GPs expressed high levels of uncertainty in their work with patients presenting problems related to mental illness or substance abuse. The complexity of the problems presented, shortage of time, limited access to reliable information and limited range of interventions available during out-of-hours contributed to this uncertainty. Perceived access to second opinion seemed to have a major impact on subjectively experienced work stress. CONCLUSIONS: The GPs experienced out-of hours psychiatry as a field with high levels of uncertainty and limited support to help them meet the experienced challenges. This might influence the quality of care provided. If the current organisation of emergency mental healthcare is to be kept, we need to provide GPs with a better support framework out-of-hours. PMID- 21619609 TI - Predictors of low cervical cancer screening among immigrant women in Ontario, Canada. AB - BACKGROUND: Disparities in cervical cancer screening are known to exist in Ontario, Canada for foreign-born women. The relative importance of various barriers to screening may vary across ethnic groups. This study aimed to determine how predictors of low cervical cancer screening, reflective of sociodemographics, the health care system, and migration, varied by region of origin for Ontario's immigrant women. METHODS: Using a validated billing code algorithm, we determined the proportion of women who were not screened during the three-year period of 2006-2008 among 455,864 identified immigrant women living in Ontario's urban centres. We created eight identical multivariate Poisson models, stratified by eight regions of origin for immigrant women. In these models, we adjusted for various sociodemographic, health care-related and migration-related variables. We then used the resulting adjusted relative risks to calculate population-attributable fractions for each variable by region of origin. RESULTS: Region of origin was not a significant source of effect modification for lack of recent cervical cancer screening. Certain variables were significantly associated with lack of screening across all or nearly all world regions. These consisted of not being in the 35-49 year age group, residence in the lowest-income neighbourhoods, not being in a primary care patient enrolment model, a provider from the same region, and not having a female provider. For all women, the highest population-attributable risk was seen for not having a female provider, with values ranging from 16.8% [95% CI 14.6-19.1%] among women from the Middle East and North Africa to 27.4% [95% CI 26.2-28.6%] for women from East Asia and the Pacific. CONCLUSIONS: To increase screening rates across immigrant groups, efforts should be made to ensure that women have access to a regular source of primary care, and ideally access to a female health professional. Efforts should also be made to increase the enrolment of immigrant women in new primary care patient enrolment models. PMID- 21619610 TI - Patient-Reported Outcome questionnaires for hip arthroscopy: a systematic review of the psychometric evidence. AB - BACKGROUND: Hip arthroscopies are often used in the treatment of intra-articular hip injuries. Patient-reported outcomes (PRO) are an important parameter in evaluating treatment. It is unclear which PRO questionnaires are specifically available for hip arthroscopy patients. The aim of this systematic review was to investigate which PRO questionnaires are valid and reliable in the evaluation of patients undergoing hip arthroscopy. METHODS: A search was conducted in Pubmed, Medline, CINAHL, the Cochrane Library, Pedro, EMBASE and Web of Science from 1931 to October 2010. Studies assessing the quality of PRO questionnaires in the evaluation of patients undergoing hip arthroscopy were included. The quality of the questionnaires was evaluated by the psychometric properties of the outcome measures. The quality of the articles investigating the questionnaires was assessed by the COSMIN list. RESULTS: Five articles identified three questionnaires; the Modified Harris Hip Score (MHHS), the Nonarthritic Hip Score (NAHS) and the Hip Outcome Score (HOS). The NAHS scored best on the content validity, whereas the HOS scored best on agreement, internal consistency, reliability and responsiveness. The quality of the articles describing the HOS scored highest. The NAHS is the best quality questionnaire. The articles describing the HOS are the best quality articles. CONCLUSIONS: This systematic review shows that there is no conclusive evidence for the use of a single patient reported outcome questionnaire in the evaluation of patients undergoing hip arthroscopy. Based on available psychometric evidence we recommend using a combination of the NAHS and the HOS for patients undergoing hip arthroscopy. PMID- 21619611 TI - Identifier mapping performance for integrating transcriptomics and proteomics experimental results. AB - BACKGROUND: Studies integrating transcriptomic data with proteomic data can illuminate the proteome more clearly than either separately. Integromic studies can deepen understanding of the dynamic complex regulatory relationship between the transcriptome and the proteome. Integrating these data dictates a reliable mapping between the identifier nomenclature resultant from the two high throughput platforms. However, this kind of analysis is well known to be hampered by lack of standardization of identifier nomenclature among proteins, genes, and microarray probe sets. Therefore data integration may also play a role in critiquing the fallible gene identifications that both platforms emit. RESULTS: We compared three freely available internet-based identifier mapping resources for mapping UniProt accessions (ACCs) to Affymetrix probesets identifications (IDs): DAVID, EnVision, and NetAffx. Liquid chromatography-tandem mass spectrometry analyses of 91 endometrial cancer and 7 noncancer samples generated 11,879 distinct ACCs. For each ACC, we compared the retrieval sets of probeset IDs from each mapping resource. We confirmed a high level of discrepancy among the mapping resources. On the same samples, mRNA expression was available. Therefore, to evaluate the quality of each ACC-to-probeset match, we calculated proteome-transcriptome correlations, and compared the resources presuming that better mapping of identifiers should generate a higher proportion of mapped pairs with strong inter-platform correlations. A mixture model for the correlations fitted well and supported regression analysis, providing a window into the performance of the mapping resources. The resources have added and dropped matches over two years, but their overall performance has not changed. CONCLUSIONS: The methods presented here serve to achieve concrete context specific insight, to support well-informed decisions in choosing an ID mapping strategy for "omic" data merging. PMID- 21619612 TI - Prothrombotic profile in patients with vasospastic or non vasospastic angina and non significant coronary stenosis. AB - BACKGROUND: Patients with vasospastic (VA) or non vasospastic angina (NVA) without significant coronary stenosis have a reduced risk of infarction but is unclear whether or not this may be attributable to a lack of prothrombotic profile - similar to that present in patients with stable coronary artery disease (CAD). METHODS: Plasma levels of von Willebrand factor, total and free tissue factor pathway inhibitor, plasminogen activator inhibitor-1, and fibrinogen were analyzed in 15 patients with stable VA and 23 with NVA, all with vasoconstrictive response to acetylcholine although with different severity. Results were compared with those of 20 age-matched controls and 10 patients with CAD. RESULTS: Plasma levels of von Willebrand factor in patients with VA or NVA were higher than in controls (207 +/- 62 and 203 +/- 69% vs 121 +/- 38%, p < 0.001) and tended to be lower than in CAD patients (264 +/- 65, p = 0.145). They also presented higher total tissue factor pathway inhibitor (123 +/- 18 and 111 +/- 25 vs 88 +/- 14, ng/ml p < 0.001) and plasminogen activator inhibitor-1 levels than controls (51 +/- 30 and 52 +/- 31% vs 19 +/- 9 ng/ml, p < 0.001) and similar to CAD patients (134 +/- 23 and 62 +/- 31, respectively, ns). Moreover, free tissue factor pathway inhibitor plasma levels were lower than controls (18 +/- 5 and 17 +/- 5 vs 23 +/- 8 ng/ml, p = 0.002) and similar to CAD patients (14 +/- 5, ns). Despite this prothrombotic condition none of VA or NVA patients presented a myocardial infarction during a 9 year follow-up, an observation also reported in larger series. CONCLUSIONS: During a stable phase of their disease, patients with VA or NVA present a prothrombotic profile that might eventually contribute to occurrence of myocardial infarction. The rarity of these events, however, may suggests that ill defined factors would protect these patients from coronary plaque rupture/fissure. PMID- 21619613 TI - Association between biomass fuel use and maternal report of child size at birth- an analysis of 2005-06 India Demographic Health Survey data. AB - BACKGROUND: Observational epidemiological studies and a systematic review have consistently shown an association between maternal exposure to biomass smoke and reduced birth weight. Our aim was to further test this hypothesis. METHODS: We analysed the data from 47,139 most recent singleton births during preceding five years of 2005-06 India Demographic Health Survey (DHS). Information about birth weight from child health card and/or mothers' recall) was analysed. Since birth weight was not recorded for nearly 60% of the reported births, maternal self report of child's size at birth was used as a proxy. Fuel type was classified as high pollution fuels (wood, straw, animal dung, and crop residues kerosene, coal and charcoal), and low pollution fuels (electricity, liquid petroleum gas (LPG), natural gas and biogas). Univariate and multivariable logistic regression models were developed using SURVEYLOGISTIC procedure in SAS system. We used three logistic regression models in which child factors, maternal factors and demographic factors were added step-by-step to the main exposure variable. Adjusted Odds Ratios (AORs) and their 95% CI were calculated. A p-value less than 0.05 was considered as significant. RESULTS: Child's birth weight was available for only 19,270 (41%) births; 3113 from health card and 16,157 from mothers' recall. For available data, mean birth weight was 2846.5 grams (SD = 684.6). Children born in households using high pollution fuels were 73 grams lighter than those born in households using low pollution fuels (mean birth weight 2883.8 grams versus 2810.7 grams, p < 0.001). Use of biomass fuels was associated with size at birth. Unadjusted OR was 1.41 (95% CI, 1.27 1.55). Adjusted OR after controlling for child factors was 1.41 (95% CI 1.29, 1.57). AOR after controlling for both child and maternal factors was 1.21 (95% CI 1.06, 1.32). In final model AOR was 1.07 (95% 0.94, 1.22) after controlling for child, maternal and demographic factors. Gender, birth order, mother's BMI, haemoglobin level and education were significant in all three models. CONCLUSIONS: Use of biomass fuels is associated with child size at birth. Future studies should investigate this association using more direct methods for measurement of exposure to smoke emitted from biomass fuels and birth weight. PMID- 21619614 TI - Inhibition of TRPA1 channel activity in sensory neurons by the glial cell line derived neurotrophic factor family member, artemin. AB - BACKGROUND: The transient receptor potential (TRP) channel subtype A1 (TRPA1) is known to be expressed on sensory neurons and respond to changes in temperature, pH and local application of certain noxious chemicals such as allyl isothiocyanate (AITC). Artemin is a neuronal survival and differentiation factor and belongs to the glial cell line-derived neurotrophic factor (GDNF) family. Both TRPA1 and artemin have been reported to be involved in pathological pain initiation and maintenance. In the present study, using whole-cell patch clamp recording technique, in situ hybridization and behavioral analyses, we examined the functional interaction between TRPA1 and artemin. RESULTS: We found that 85.8 +/- 1.9% of TRPA1-expressing neurons also expressed GDNF family receptor alpha 3 (GFR alpha3), and 87.5 +/- 4.1% of GFRalpha3-expressing neurons were TRPA1 positive. In whole-cell patch clamp analysis, a short-term treatment of 100 ng/ml artemin significantly suppressed the AITC-induced TRPA1 currents. A concentration response curve of AITC resulting from the effect of artemin showed that this inhibition did not change EC50 but did lower the AITC-induced maximum response. In addition, pre-treatment of artemin significantly suppressed the number of paw lifts induced by intraplantar injection of AITC, as well as the formalin-induced pain behaviors. CONCLUSIONS: These findings that a short-term application of artemin inhibits the TRPA1 channel's activity and the sequential pain behaviors suggest a role of artemin in regulation of sensory neurons. PMID- 21619615 TI - GPCR oligomers in pharmacology and signaling. AB - G protein-coupled receptors (GPCRs) represent one of the largest families of cell surface receptors, and are the target of more than half of the current therapeutic drugs on the market. When activated by an agonist, the GPCR undergoes conformational changes that facilitate its interaction with heterotrimeric G proteins, which then relay signals to downstream intracellular effectors. Although GPCRs were thought to function as monomers, many studies support the hypothesis that G protein coupling involves the formation of GPCR homo- and/or hetero-complexes. These complex systems have been suggested to exhibit specific signaling cascades, pharmacological, internalization, and recycling properties. In this review, we summarize recent advances in our understanding of the structure, function and dynamics of GPCR complexes, as well as the findings obtained in animal models. PMID- 21619617 TI - A deterministic map of Waddington's epigenetic landscape for cell fate specification. AB - BACKGROUND: The image of the "epigenetic landscape", with a series of branching valleys and ridges depicting stable cellular states and the barriers between those states, has been a popular visual metaphor for cell lineage specification - especially in light of the recent discovery that terminally differentiated adult cells can be reprogrammed into pluripotent stem cells or into alternative cell lineages. However the question of whether the epigenetic landscape can be mapped out quantitatively to provide a predictive model of cellular differentiation remains largely unanswered. RESULTS: Here we derive a simple deterministic path integral quasi-potential, based on the kinetic parameters of a gene network regulating cell fate, and show that this quantity is minimized along a temporal trajectory in the state space of the gene network, thus providing a marker of directionality for cell differentiation processes. We then use the derived quasi potential as a measure of "elevation" to quantitatively map the epigenetic landscape, on which trajectories flow "downhill" from any location. Stochastic simulations confirm that the elevation of this computed landscape correlates to the likelihood of occurrence of particular cell fates, with well-populated low lying "valleys" representing stable cellular states and higher "ridges" acting as barriers to transitions between the stable states. CONCLUSIONS: This quantitative map of the epigenetic landscape underlying cell fate choice provides mechanistic insights into the "forces" that direct cellular differentiation in the context of physiological development, as well as during artificially induced cell lineage reprogramming. Our generalized approach to mapping the landscape is applicable to non-gradient gene regulatory systems for which an analytical potential function cannot be derived, and also to high-dimensional gene networks. Rigorous quantification of the gene regulatory circuits that govern cell lineage choice and subsequent mapping of the epigenetic landscape can potentially help identify optimal routes of cell fate reprogramming. PMID- 21619616 TI - Transcriptional dysregulation of 5-HT1A autoreceptors in mental illness. AB - The serotonin-1A (5-HT1A) receptor is among the most abundant and widely distributed 5-HT receptors in the brain, but is also expressed on serotonin neurons as an autoreceptor where it plays a critical role in regulating the activity of the entire serotonin system. Over-expression of the 5-HT1A autoreceptor has been implicated in reducing serotonergic neurotransmission, and is associated with major depression and suicide. Extensive characterization of the transcriptional regulation of the 5-HT1A gene (HTR1A) using cell culture systems has revealed a GC-rich "housekeeping" promoter that non-selectively drives its expression; this is flanked by a series of upstream repressor elements for REST, Freud-1/CC2D1A and Freud-2/CC2D1B factors that not only restrict its expression to neurons, but may also regulate the level of expression of 5-HT1A receptors in various subsets of neurons, including serotonergic neurons. A separate set of allele-specific factors, including Deaf1, Hes1 and Hes5 repress at the HTR1A C(-1019)G (rs6295) polymorphism in serotonergic neurons in culture, as well as in vivo. Pet1, an obligatory enhancer for serotonergic differentiation, has been identified as a potent activator of 5-HT1A autoreceptor expression. Taken together, these results highlight an integrated regulation of 5 HT1A autoreceptors that differs in several aspects from regulation of post synaptic 5-HT1A receptors, and could be selectively targeted to enhance serotonergic neurotransmission. PMID- 21619618 TI - Adaptation to altered balance conditions in unilateral amputees due to atherosclerosis: a randomized controlled study. AB - BACKGROUND: Amputation impairs the ability to balance. We examined adaptation strategies in balance following dysvascularity-induced unilateral tibial amputation in skilled prosthetic users (SPU) and first fitted amputees (FFA) (N = 28). METHODS: Excursions of center of pressure (COP) were determined during 20 s quiet standing using a stabilometry system with eyes-open on both legs or on the non-affected leg(s). MAIN MEASURES: COP trajectories and time functions; distribution of reaction forces between the two legs; inclination angles obtained through second order regression analysis using stabilogram data. RESULTS: FFA vs SPU demonstrated 27.8% greater postural sway in bilateral stance (p = 0.0004). Postural sway area was smaller in FFA standing on the non-affected leg compared with SPU (p = 0.028). The slope of the regression line indicating postural stability was nearly identical in FFA and SPU and the direction of regression line was opposite for the left and right leg amputees. CONCLUSION: Of the two adaptation strategies in balance, the first appears before amputation due to pain and fatigue in the affected leg. This strategy appears in the form of reduced postural sway while standing on the non-affected leg. The second adaptation occurs during rehabilitation and regular use of the prosthesis resulting in normal weightbearing associated with reduced postural sway on two legs and return to the normal postural stability on one leg. PMID- 21619619 TI - Comparisons of 7- to 78-joint ultrasonography scores: all different joint combinations show equal response to adalimumab treatment in patients with rheumatoid arthritis. AB - INTRODUCTION: The primary objectives were to explore the associations between a comprehensive ultrasonographic (US) assessment of joints, tendons and bursae and previously described reduced joint counts (7-, 12-, 28- and 44-joint score) as well as to assess the sensitivity to change of these different US joint combinations during biological treatment. METHODS: Twenty patients with rheumatoid arthritis (RA) were examined by US (B-mode (BM) and power Doppler (PD)) with use of a semi-quantitative (0 to 3) score of 78 joints, 36 tendons/tendon groups and two bursae (hereafter described as the 78-joint score) at baseline and 1, 3, 6 and 12 months after initiating treatment with adalimumab. BM and PD scores for the different joint combinations were generated. RESULTS: The reduced joint scores had high correlation coefficients with the 78-joint score at all examinations (range 0.79 to 0.99 for BM and 0.77 to 0.99 for PD, each P < 0.001) and sum BM and PD scores of all the different joint combinations improved significantly during follow-up (P <= 0.05 to 0.001). CONCLUSIONS: The reduced joint combinations were highly associated to the 78-joint score. Furthermore, all the joint combinations presently explored responded well to biological treatment. This indicates that an approach focusing on few joints and tendons gives equivalent information about the inflammatory activity in RA patients as a comprehensive US examination. The optimal combination of joints and tendons for a valid, reliable and feasible US measurement should be further explored to define a US score for follow-up of RA patients on biological treatment. PMID- 21619620 TI - Videoradiographic analysis of the range of motion in unilateral experimental knee joint arthritis in rats. AB - INTRODUCTION: The translational and predictive value of animal models highly depends on the validity of respective readout parameters. In arthritis research, there has been a shift from sole threshold testing for pain-related behavior, as well as from swelling and histology assessment for inflammation, toward an analysis of joint function as indicated, for instance, by an increasing number of studies on gait abnormalities. Clinically, the range of motion (ROM) of the affected joint plays a major role in diagnosis and the assessment of treatment benefits. This parameter, however, is only insufficiently detected by currently used analytic systems in animals. METHODS: Here we used high-resolution videoradiographic analysis to assess ROM in experimental knee joint arthritis in rats. This parameter is described during the 21-day course of antigen-induced arthritis in rats. Furthermore, the therapeutic effects of antinociceptive (morphine) and anti-inflammatory (dexamethasone) treatment on ROM are documented. To obtain additional information on the implications of ROM in animal models, correlations were performed to measure pain-related behavior and inflammation. RESULTS: The study animals showed a significant reduction in ROM of the inflamed knee joint in the acute phase of arthritis. This was accompanied by an increase in knee joint movement on the contralateral side, indicating a compensational mechanism. Both morphine and dexamethasone treatment increased and thus normalized ROM. Changes in ROM were further stage-dependently correlated with weight bearing and joint swelling, that is, with both pain-related behavior and signs of inflammation. CONCLUSIONS: The dynamic ROM observed in freely moving rats in our model of knee joint arthritis might serve as a parameter for global disease activity and might thus represent a promising readout parameter for preclinical assessment regarding the overall efficacy not only of antiarthritic but also of antinociceptive compounds. PMID- 21619621 TI - Effects of an evidence service on health-system policy makers' use of research evidence: a protocol for a randomised controlled trial. AB - BACKGROUND: Health-system policy makers need timely access to synthesised research evidence to inform the policy-making process. No efforts to address this need have been evaluated using an experimental quantitative design. We developed an evidence service that draws inputs from Health Systems Evidence, which is a database of policy-relevant systematic reviews. The reviews have been (a) categorised by topic and type of review; (b) coded by the last year searches for studies were conducted and by the countries in which included studies were conducted; (c) rated for quality; and (d) linked to available user-friendly summaries, scientific abstracts, and full-text reports. Our goal is to evaluate whether a "full-serve" evidence service increases the use of synthesized research evidence by policy analysts and advisors in the Ontario Ministry of Health and Long-Term Care (MOHLTC) as compared to a "self-serve" evidence service. METHODS/DESIGN: We will conduct a two-arm randomized controlled trial (RCT), along with a follow-up qualitative process study in order to explore the findings in greater depth. For the RCT, all policy analysts and policy advisors (n = 168) in a single division of the MOHLTC will be invited to participate. Using a stratified randomized design, participants will be randomized to receive either the "full-serve" evidence service (database access, monthly e-mail alerts, and full-text article availability) or the "self-serve" evidence service (database access only). The trial duration will be ten months (two-month baseline period, six-month intervention period, and two month cross-over period). The primary outcome will be the mean number of site visits/month/user between baseline and the end of the intervention period. The secondary outcome will be participants' intention to use research evidence. For the qualitative study, 15 participants from each trial arm (n = 30) will be purposively sampled. One-on-one semi structured interviews will be conducted by telephone on their views about and their experiences with the evidence service they received, how helpful it was in their work, why it was helpful (or not helpful), what aspects were most and least helpful and why, and recommendations for next steps. DISCUSSION: To our knowledge, this will be the first RCT to evaluate the effects of an evidence service specifically designed to support health-system policy makers in finding and using research evidence. TRIAL REGISTRATION: ClinicalTrials.gov: NCT01307228. PMID- 21619622 TI - Effects of an evidence service on community-based AIDS service organizations' use of research evidence: a protocol for a randomized controlled trial. AB - BACKGROUND: To support the use of research evidence by community-based organizations (CBOs) we have developed 'Synthesized HIV/AIDS Research Evidence' (SHARE), which is an evidence service for those working in the HIV sector. SHARE consists of several components: an online searchable database of HIV-relevant systematic reviews (retrievable based on a taxonomy of topics related to HIV/AIDS and open text search); periodic email updates; access to user-friendly summaries; and peer relevance assessments. Our objective is to evaluate whether this 'full serve' evidence service increases the use of research evidence by CBOs as compared to a 'self-serve' evidence service. METHODS/DESIGN: We will conduct a two-arm randomized controlled trial (RCT), along with a follow-up qualitative process study to explore the findings in greater depth. All CBOs affiliated with Canadian AIDS Society (n = 120) will be invited to participate and will be randomized to receive either the 'full-serve' version of SHARE or the 'self serve' version (a listing of relevant systematic reviews with links to records on PubMed and worksheets that help CBOs find and use research evidence) using a simple randomized design. All management and staff from each organization will be provided access to the version of SHARE that their organization is allocated to. The trial duration will be 10 months (two-month baseline period, six-month intervention period, and two month crossover period), the primary outcome measure will be the mean number of logins/month/organization (averaged across the number of users from each organization) between baseline and the end of the intervention period. The secondary outcome will be intention to use research evidence as measured by a survey administered to one key decision maker from each organization. For the qualitative study, one key organizational decision maker from 15 organizations in each trial arm (n = 30) will be purposively sampled. One on-one semi-structured interviews will be conducted by telephone on their views about and their experiences with the evidence service they received, how helpful it was in their work, why it was helpful (or not helpful), what aspects were most and least helpful and why, and recommendations for next steps. DISCUSSION: To our knowledge, this will be the first RCT to evaluate the effects of an evidence service specifically designed to support CBOs in finding and using research evidence. TRIAL REGISTRATION: ClinicalTrials.gov: NCT01257724. PMID- 21619623 TI - A miRNA expression signature that separates between normal and malignant prostate tissues. AB - BACKGROUND: MicroRNAs (miRNAs) constitute a class of small non-coding RNAs that post-transcriptionally regulate genes involved in several key biological processes and thus are involved in various diseases, including cancer. In this study we aimed to identify a miRNA expression signature that could be used to separate between normal and malignant prostate tissues. RESULTS: Nine miRNAs were found to be differentially expressed (p <0.00001). With the exception of two samples, this expression signature could be used to separate between the normal and malignant tissues. A cross-validation procedure confirmed the generality of this expression signature. We also identified 16 miRNAs that possibly could be used as a complement to current methods for grading of prostate tumor tissues. CONCLUSIONS: We found an expression signature based on nine differentially expressed miRNAs that with high accuracy (85%) could classify the normal and malignant prostate tissues in patients from the Swedish Watchful Waiting cohort. The results show that there are significant differences in miRNA expression between normal and malignant prostate tissue, indicating that these small RNA molecules might be important in the biogenesis of prostate cancer and potentially useful for clinical diagnosis of the disease. PMID- 21619624 TI - High frequency of Plasmodium falciparum chloroquine resistance marker (pfcrt T76 mutation) in Yemen: an urgent need to re-examine malaria drug policy. AB - BACKGROUND: Malaria remains a significant health problem in Yemen with Plasmodium falciparum being the predominant species which is responsible for 90% of the malaria cases. Despite serious concerns regarding increasing drug resistance, chloroquine is still used for the prevention and treatment of malaria in Yemen. This study was carried out to determine the prevalence of choloroquine resistance (CQR) of P. falciparum isolated from Yemen based on the pfcrt T76 mutation. METHODS: A cross-sectional study was carried out among 511 participants from four governorates in Yemen. Blood samples were screened using microscopic and species specific nested PCR based on the 18S rRNA gene to detect and identify Plasmodium species. Blood samples positive for P. falciparum were used for detecting the pfcrt T76 mutation using nested-PCR. RESULTS: The prevalence of pfcrt T76 mutation was 81.5% (66 of 81 isolates). Coastal areas/foothills had higher prevalence of pfcrt T76 mutation compared to highland areas (90.5% vs 71.8%) (p = 0.031). The pfcrt T76 mutation had a significant association with parasitaemia (p = 0.045). Univariate analysis shows a significant association of pfcrt T76 mutation with people aged > 10 years (OR = 9, 95% CI = 2.3 - 36.2, p = 0.001), low household income (OR = 5, 95% CI = 1.3 - 19.5, p = 0.027), no insecticide spray (OR = 3.7, 95% CI = 1.16 - 11.86, p = 0.025) and not sleeping under insecticide treated nets (ITNs) (OR = 4.8, 95% CI = 1.38 - 16.78, p = 0.01). Logistic regression model confirmed age > 10 years and low household income as predictors of pfcrt T76 mutation in Yemen P. falciparum isolates. CONCLUSIONS: The high prevalence of pfcrt T76 mutation in Yemen could be a predictive marker for the prevalence of P. falciparum CQR. This finding shows the necessity for an in-vivo therapeutic efficacy test for CQ. P. falciparum CQR should be addressed in the national strategy to control malaria. PMID- 21619625 TI - Endovascular treatment of iatrogenic axillary artery pseudoaneurysm under echographic control: a case report. AB - AIM: Brief case report of the treatment of a large axillary artery pseudoaneurysm after a pacemaker using a left brachial cutdown and a retrograde delivery of a covered stent using ultrasound and fluoroscopic guidance. The patient's renal function precluded the use of contrast materials. CASE REPORT: A 77 years old man presenting with acute renal failure and haemoglobin decrease arrived with an expanding pseudoaneurysm of the left axillary artery from a pacemaker placement. Considering the site of the lesion and patient's comorbidities, under echographic control, a Hemobahn(r) stent-graft was placed; fluoroscopy assisted manipulation of guidewires and sheaths into the aortic arch. The procedure was successfully ended without any complications. At 8 months the stent graft was still patent. CONCLUSION: Ultrasound guidance may represent an alternative for pseudo-aneurysm exclusion without any use of contrast medium, especially in those patient where lesions are easily detectable using ultrasonography and when comorbidities contraindicate aggressive surgical or angiographic approach. PMID- 21619626 TI - The CRE1 carbon catabolite repressor of the fungus Trichoderma reesei: a master regulator of carbon assimilation. AB - BACKGROUND: The identification and characterization of the transcriptional regulatory networks governing the physiology and adaptation of microbial cells is a key step in understanding their behaviour. One such wide-domain regulatory circuit, essential to all cells, is carbon catabolite repression (CCR): it allows the cell to prefer some carbon sources, whose assimilation is of high nutritional value, over less profitable ones. In lower multicellular fungi, the C2H2 zinc finger CreA/CRE1 protein has been shown to act as the transcriptional repressor in this process. However, the complete list of its gene targets is not known. RESULTS: Here, we deciphered the CRE1 regulatory range in the model cellulose and hemicellulose-degrading fungus Trichoderma reesei (anamorph of Hypocrea jecorina) by profiling transcription in a wild-type and a delta-cre1 mutant strain on glucose at constant growth rates known to repress and de-repress CCR-affected genes. Analysis of genome-wide microarrays reveals 2.8% of transcripts whose expression was regulated in at least one of the four experimental conditions: 47.3% of which were repressed by CRE1, whereas 29.0% were actually induced by CRE1, and 17.2% only affected by the growth rate but CRE1 independent. Among CRE1 repressed transcripts, genes encoding unknown proteins and transport proteins were overrepresented. In addition, we found CRE1-repression of nitrogenous substances uptake, components of chromatin remodeling and the transcriptional mediator complex, as well as developmental processes. CONCLUSIONS: Our study provides the first global insight into the molecular physiological response of a multicellular fungus to carbon catabolite regulation and identifies several not yet known targets in a growth-controlled environment. PMID- 21619627 TI - Transcriptome instability in colorectal cancer identified by exon microarray analyses: Associations with splicing factor expression levels and patient survival. AB - BACKGROUND: Colorectal cancer (CRC) is a heterogeneous disease that, on the molecular level, can be characterized by inherent genomic instabilities; chromosome instability and microsatellite instability. In the present study we analyze genome-wide disruption of pre-mRNA splicing, and propose transcriptome instability as a characteristic that is analogous to genomic instability on the transcriptome level. METHODS: Exon microarray profiles from two independent series including a total of 160 CRCs were investigated for their relative amounts of exon usage differences. Each exon in each sample was assigned an alternative splicing score calculated by the FIRMA algorithm. Amounts of deviating exon usage per sample were derived from exons with extreme splicing scores. RESULTS: There was great heterogeneity within both series in terms of sample-wise amounts of deviating exon usage. This was strongly associated with the expression levels of approximately half of 280 splicing factors (54% and 48% of splicing factors were significantly correlated to deviating exon usage amounts in the two series). Samples with high or low amounts of deviating exon usage, associated with overall transcriptome instability, were almost completely separated into their respective groups by hierarchical clustering analysis of splicing factor expression levels in both sample series. Samples showing a preferential tendency towards deviating exon skipping or inclusion were associated with skewed transcriptome instability. There were significant associations between transcriptome instability and reduced patient survival in both sample series. In the test series, patients with skewed transcriptome instability showed the strongest prognostic association (P = 0.001), while a combination of the two characteristics showed the strongest association with poor survival in the validation series (P = 0.03). CONCLUSIONS: We have described transcriptome instability as a characteristic of CRC. This transcriptome instability has associations with splicing factor expression levels and poor patient survival. PMID- 21619628 TI - Binding of ATP to vascular endothelial growth factor isoform VEGF-A165 is essential for inducing proliferation of human umbilical vein endothelial cells. AB - BACKGROUND: ATP binding is essential for the bioactivity of several growth factors including nerve growth factor, fibroblast growth factor-2 and brain derived neurotrophic factor. Vascular endothelial growth factor isoform 165 (VEGF A(165)) induces the proliferation of human umbilical vein endothelial cells, however a dependence on ATP-binding is currently unknown. The aim of the present study was to determine if ATP binding is essential for the bioactivity of VEGF A(165). RESULTS: We found evidence that ATP binding to VEGF-A(165) induced a conformational change in the secondary structure of the growth factor. This binding appears to be significant at the biological level, as we found evidence that nanomolar levels of ATP (4-8 nm) are required for the VEGF-A(165)-induced proliferation of human umbilical vein endothelial cells. At these levels, purinergic signaling by ATP via P2 receptors can be excluded. Addition of alkaline phosphate to cell culture lowered the ATP concentration in the cell culture medium to 1.8 nM and inhibited cell proliferation. CONCLUSIONS: We propose that proliferation of endothelial cells is induced by a VEGF-A(165)-ATP complex, rather than VEGF-A(165) alone. PMID- 21619629 TI - Evidence for a heritable predisposition to Chronic Fatigue Syndrome. AB - BACKGROUND: Chronic Fatigue Syndrome (CFS) came to attention in the 1980s, but initial investigations did not find organic causes. Now decades later, the etiology of CFS has yet to be understood, and the role of genetic predisposition in CFS remains controversial. Recent reports of CFS association with the retrovirus xenotropic murine leukemic virus-related virus (XMRV) or other murine leukemia related retroviruses (MLV) might also suggest underlying genetic implications within the host immune system. METHODS: We present analyses of familial clustering of CFS in a computerized genealogical resource linking multiple generations of genealogy data with medical diagnosis data of a large Utah health care system. We compare pair-wise relatedness among cases to expected relatedness in the Utah population, and we estimate risk for CFS for first, second, and third degree relatives of CFS cases. RESULTS: We observed significant excess relatedness of CFS cases compared to that expected in this population. Significant excess relatedness was observed for both close (p <0.001) and distant relationships (p = 0.010). We also observed significant excess CFS relative risk among first (2.70, 95% CI: 1.56-4.66), second (2.34, 95% CI: 1.31-4.19), and third degree relatives (1.93, 95% CI: 1.21-3.07). CONCLUSIONS: These analyses provide strong support for a heritable contribution to predisposition to Chronic Fatigue Syndrome. A population of high-risk CFS pedigrees has been identified, the study of which may provide additional understanding. PMID- 21619630 TI - Phylogeny and androgenesis in the invasive Corbicula clams (Bivalvia, Corbiculidae) in Western Europe. AB - BACKGROUND: The genus Corbicula is one of the most invasive groups of molluscs. It includes both sexual and androgenetic lineages. The present study re-assessed the different morphotypes and haplotypes of West European Corbicula in order to clarify their taxonomic identification and phylogenetic relationships with American and Asian Corbicula clams. We studied several populations from West European river basins (Meuse, Seine, Rhine and Rhone) through an "integrative taxonomy" approach. We combined morphology, partial mitochondrial COI and cyt b sequences and eleven microsatellite loci. Furthermore, we looked for discrepancies between mtDNA and nrDNA/morphology, indicative of androgenesis between lineages. RESULTS: There are three Corbicula morphotypes in Western Europe associated to three mitochondrial lineages and three genotypes. Form R shares the same COI haplotype as the American form A and the Japanese C. leana. Form S and the American form C have the same haplotype, although their morphologies seem divergent. The European form Rlc belongs to the same mitochondrial lineage as both the American form B and the Asian C. fluminea.Interestingly, within each haplotype/genotype or lineage, no genetic diversity was found although their invasive success is high. Moreover, we detected rare mismatches between mtDNA and nrDNA/morphology, indicative of androgenesis and mitochondrial capture between form R and form S and therefore challenging the phylogenetic relatedness and the species status within this genus. The global phylogenetic analysis revealed that the sexual Corbicula lineages seem restricted to the native areas while their androgenetic relatives are widespread and highly invasive. CONCLUSIONS: We clarified the discrepancies and incongruent results found in the literature about the European morphotypes of Corbicula and associated mitochondrial lineages. The three West European morphotypes belong to three distinct nuclear and mitochondrial lineages. However mitochondrial capture occurs in sympatric populations of forms R and S. The species status of the morphotypes therefore remains doubtful. Moreover the androgenetic lineages seem widely distributed compared to their sexual relatives, suggesting that androgenesis and invasive success may be linked in the genus Corbicula. PMID- 21619631 TI - Complete transposition of the aorta and pulmonary artery in a Belgian Blue crossbreed calf: a case report. AB - BACKGROUND: Complete transposition of the great arteries is a congenital cardiac malformation occasionally encountered in cattle and other species. The objective of the present report was to provide a detailed clinical, echocardiographic and post mortem description of a calf presenting with this condition. CASE PRESENTATION: A 6-week old male Belgian Blue cross-breed calf was examined for respiratory distress and exercise intolerance. The patient was bright, alert and responsive without any neurologic abnormalities but was exercise intolerant, had marked cyanosis, tachycardia, tachypnea, a pansystolic heart murmur as well as a bilaterally palpable thrill over the heart. Arterial blood gas analysis revealed marked hypoxemia (PaO(2)=23 mmHg, O(2)sat=41.1%), mild hypercapnia and compensated respiratory acidosis. Echocardiographic examination revealed a complete transposition of the great arteries in combination with a ventricular septal defect through which blood shunted bidirectionally. Cardiac catheterization confirmed that arterialization of blood of the systemic circulation solely occurred in the right ventricle through blood shunting from the left into the right ventricle. Results of post mortem examination are presented. CONCLUSION: Complete transposition of the great arteries is a cyanotic congenital anomaly repeatedly reported in calves that should be considered as differential diagnosis in patients presenting with hypoxemia more severe than commonly encountered with other congenital cyanotic heart conditions. We give a comprehensive summary of the clinical presentation, diagnostic work-up and post mortem examination of a Belgian Blue cross-breed calf with complete transposition of the great arteries. PMID- 21619632 TI - Consumer purchasing patterns in response to calorie labeling legislation in New York City. AB - BACKGROUND: Obesity is a major public health threat and policies aimed at curbing this epidemic are emerging. National calorie labeling legislation is forthcoming and requires rigorous evaluation to examine its impact on consumers. The purpose of this study was to examine whether point-of-purchase calorie labels in New York City (NYC) chain restaurants affected food purchasing patterns in a sample of lower income adults in NYC and Newark, NJ. METHODS: This study utilized a difference-in-difference design to survey 1,170 adult patrons of four popular chain restaurants in NYC and Newark, NJ (which did not introduce labeling) before and after calorie labeling was implemented in NYC. Receipt data were collected and analyzed to examine food and beverage purchases and frequency of fast food consumption. Descriptive statistics were generated, and linear and logistic regression, difference-in-difference analysis, and predicted probabilities were used to analyze the data. RESULTS: A difference-in-difference analysis revealed no significant favorable differences and some unfavorable differences in food purchasing patterns and frequency of fast food consumption between adult patrons of fast food restaurants in NYC and Newark, NJ. Adults in NYC who reported noticing and using the calorie labels consumed fast food less frequently compared to adults who did not notice the labels (4.9 vs. 6.6 meals per week, p <0.05). CONCLUSION: While no favorable differences in purchasing as a result of labeling were noted, self-reported use of calorie labels was associated with some favorable behavioral patterns in a subset of adults in NYC. However, overall impact of the legislation may be limited. More research is needed to understand the most effective way to deliver calorie information to consumers. PMID- 21619633 TI - Genomic features and computational identification of human microRNAs under long range developmental regulation. AB - BACKGROUND: Recent functional studies have demonstrated that many microRNAs (miRNAs) are expressed by RNA polymerase II in a specific spatiotemporal manner during the development of organisms and play a key role in cell-lineage decisions and morphogenesis. They are therefore functionally related to a number of key protein coding developmental genes, that form genomic regulatory blocks (GRBs) with arrays of highly conserved non-coding elements (HCNEs) functioning as long range enhancers that collaboratively regulate the expression of their target genes. Given this functional similarity as well as recent zebrafish transgenesis assays showing that the miR-9 family is indeed regulated by HCNEs with enhancer activity, we hypothesized that this type of miRNA regulation is prevalent. In this paper, we therefore systematically investigate the regulatory landscape around conserved self-transcribed miRNAs (ST miRNAs), with their own known or computationally inferred promoters, by analyzing the hallmarks of GRB target genes. These include not only the density of HCNEs in their vicinity but also the presence of large CpG islands (CGIs) and distinct patterns of histone modification marks associated with developmental genes. RESULTS: Our results show that a subset of the conserved ST miRNAs we studied shares properties similar to those of protein-coding GRB target genes: they are located in regions of significantly higher HCNE/enhancer binding density and are more likely to be associated with CGIs. Furthermore, their putative promoters have both activating as well as silencing histone modification marks during development and differentiation. Based on these results we used both an elevated HCNE density in the genomic vicinity as well as the presence of a bivalent promoter to identify 29 putative GRB target miRNAs/miRNA clusters, over two-thirds of which are known to play a role during development and differentiation. Furthermore these predictions include miRNAs of the miR-9 family, which are the only experimentally verified GRB target miRNAs. CONCLUSIONS: A subset of the conserved miRNA loci we investigated exhibits typical characteristics of GRB target genes, which may partially explain their complex expression profiles during development. PMID- 21619634 TI - Nonlinear observer output-feedback MPC treatment scheduling for HIV. AB - BACKGROUND: Mathematical models of the immune response to the Human Immunodeficiency Virus demonstrate the potential for dynamic schedules of Highly Active Anti-Retroviral Therapy to enhance Cytotoxic Lymphocyte-mediated control of HIV infection. METHODS: In previous work we have developed a model predictive control (MPC) based method for determining optimal treatment interruption schedules for this purpose. In this paper, we introduce a nonlinear observer for the HIV-immune response system and an integrated output-feedback MPC approach for implementing the treatment interruption scheduling algorithm using the easily available viral load measurements. We use Monte-Carlo approaches to test robustness of the algorithm. RESULTS: The nonlinear observer shows robust state tracking while preserving state positivity both for continuous and discrete measurements. The integrated output-feedback MPC algorithm stabilizes the desired steady-state. Monte-Carlo testing shows significant robustness to modeling error, with 90% success rates in stabilizing the desired steady-state with 15% variance from nominal on all model parameters. CONCLUSIONS: The possibility of enhancing immune responsiveness to HIV through dynamic scheduling of treatment is exciting. Output-feedback Model Predictive Control is uniquely well-suited to solutions of these types of problems. The unique constraints of state positivity and very slow sampling are addressable by using a special-purpose nonlinear state estimator, as described in this paper. This shows the possibility of using output-feedback MPC based algorithms for this purpose. PMID- 21619635 TI - The Yin and Yang actions of North American ginseng root in modulating the immune function of macrophages. AB - BACKGROUND: Immuno-modulatory effects of ginseng, including both immuno stimulatory and immuno-suppressive effects, have been widely reported. This study aims to determine whether the paradoxical immuno-modulatory effect is related to unique phytochemical profiles of different North American (NA) ginseng, namely aqueous (AQ) and alcoholic (ALC) extracts. METHODS: AQ and ALC extracts were prepared and their immuno-bioactivity were studied in vitro in murine macrophages (Raw 264.7) through measuring the direct stimulatory production of pro inflammatory mediator and cytokines as well as the suppression of lipopolysaccharide (LPS)-stimulatory response by the two extracts. Gel permeation chromatography was used to fractionate and isolate phytochemicals for characterization of ginseng extracts. RESULTS: AQ extract up-regulated the production of nitric oxide (NO), tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) while ALC extract did not. ALC extract but not AQ extract suppressed LPS-induced macrophage NO and TNF-alpha production. These immuno stimulatory and suppressive effects were exhibited at similar extract concentrations. Moreover, the macrophage-stimulating activity of the AQ extract was inhibited in the presence of ALC extract. Fractionation of AQ extract revealed the presence of two major peaks at 230 nm with average molecular weights of 73,000 and 37,000 Da. The first fraction had similar elution volume as the crude polysaccharide (PS) fraction isolated from the AQ extract, and it was the only bioactive species. Parallel fractionation study of ALC extract yielded similar elution profiles; however, both sub-fractions were devoid of PS. Fraction I of the ALC extract suppressed LPS-induced NO production dose-dependently. CONCLUSION: ALC extract of NA ginseng, which was devoid of PS, was immuno inhibitory whereas the AQ extract, which contained PS, was immuno-stimulatory. These extract-related anti-inflammatory and pro-inflammatory effects may be considered as the Yin and Yang actions of ginseng. PMID- 21619636 TI - Understanding tumor heterogeneity as functional compartments--superorganisms revisited. AB - Compelling evidence broadens our understanding of tumors as highly heterogeneous populations derived from one common progenitor. In this review we portray various stages of tumorigenesis, tumor progression, self-seeding and metastasis in analogy to the superorganisms of insect societies to exemplify the highly complex architecture of a neoplasm as a system of functional "castes."Accordingly, we propose a model in which clonal expansion and cumulative acquisition of genetic alterations produce tumor compartments each equipped with distinct traits and thus distinct functions that cooperate to establish clinically apparent tumors. This functional compartment model also suggests mechanisms for the self construction of tumor stem cell niches. Thus, thinking of a tumor as a superorganism will provide systemic insight into its functional compartmentalization and may even have clinical implications. PMID- 21619637 TI - Transcriptome profiling of chemosensory appendages in the malaria vector Anopheles gambiae reveals tissue- and sex-specific signatures of odor coding. AB - BACKGROUND: Chemosensory signal transduction guides the behavior of many insects, including Anopheles gambiae, the major vector for human malaria in sub-Saharan Africa. To better understand the molecular basis of mosquito chemosensation we have used whole transcriptome RNA sequencing (RNA-seq) to compare transcript expression profiles between the two major chemosensory tissues, the antennae and maxillary palps, of adult female and male An. gambiae. RESULTS: We compared chemosensory tissue transcriptomes to whole body transcriptomes of each sex to identify chemosensory enhanced genes. In the six data sets analyzed, we detected expression of nearly all known chemosensory genes and found them to be highly enriched in both olfactory tissues of males and females. While the maxillary palps of both sexes demonstrated strict chemosensory gene expression overlap, we observed acute differences in sensory specialization between male and female antennae. The relatively high expression levels of chemosensory genes in the female antennae reveal its role as an organ predominately assigned to chemosensation. Remarkably, the expression of these genes was highly conserved in the male antennae, but at much lower relative levels. Alternatively, consistent with a role in mating, the male antennae displayed significant enhancement of genes involved in audition, while the female enhancement of these genes was observed, but to a lesser degree. CONCLUSIONS: These findings suggest that the chemoreceptive spectrum, as defined by gene expression profiles, is largely similar in female and male An. gambiae. However, assuming sensory receptor expression levels are correlated with sensitivity in each case, we posit that male and female antennae are perceptive to the same stimuli, but possess inverse receptive prioritizations and sensitivities. Here we have demonstrated the use of RNA-seq to characterize the sensory specializations of an important disease vector and grounded future studies investigating chemosensory processes. PMID- 21619638 TI - Determination of pore size distribution at the cell-hydrogel interface. AB - BACKGROUND: Analyses of the pore size distribution in 3D matrices such as the cell-hydrogel interface are very useful when studying changes and modifications produced as a result of cellular growth and proliferation within the matrix, as pore size distribution plays an important role in the signaling and microenvironment stimuli imparted to the cells. However, the majority of the methods for the assessment of the porosity in biomaterials are not suitable to give quantitative information about the textural properties of these nano interfaces. FINDINGS: Here, we report a methodology for determining pore size distribution at the cell-hydrogel interface, and the depth of the matrix modified by cell growth by entrapped HepG(2) cells in microcapsules made of 0.8% and 1.4% w/v alginate. The method is based on the estimation of the shortest distance between two points of the fibril-like network hydrogel structures using image analysis of TEM pictures. Values of pore size distribution determined using the presented method and those obtained by nitrogen physisorption measurements were compared, showing good agreement. A combination of these methodologies and a study of the cell-hydrogel interface at various cell culture times showed that after three days of culture, HepG(2) cells growing in hydrogels composed of 0.8% w/v alginate had more coarse of pores at depths up to 40 nm inwards (a phenomenon most notable in the first 20 nm from the interface). This coarsening phenomenon was weakly observed in the case of cells cultured in hydrogels composed of 1.4% w/v alginate. CONCLUSIONS: The method purposed in this paper allows us to obtain information about the radial deformation of the hydrogel matrix due to cell growth, and the consequent modification of the pore size distribution pattern surrounding the cells, which are extremely important for a wide spectrum of biotechnological, pharmaceutical and biomedical applications. PMID- 21619639 TI - Partially observed bipartite network analysis to identify predictive connections in transcriptional regulatory networks. AB - BACKGROUND: Messenger RNA expression is regulated by a complex interplay of different regulatory proteins. Unfortunately, directly measuring the individual activity of these regulatory proteins is difficult, leaving us with only the resulting gene expression pattern as a marker for the underlying regulatory network or regulator-gene associations. Furthermore, traditional methods to predict these regulator-gene associations do not define the relative importance of each association, leading to a large number of connections in the global regulatory network that, although true, are not useful. RESULTS: Here we present a Bayesian method that identifies which known transcriptional relationships in a regulatory network are consistent with a given body of static gene expression data by eliminating the non-relevant ones. The Partially Observed Bipartite Network (POBN) approach developed here is tested using E. coli expression data and a transcriptional regulatory network derived from RegulonDB. When the regulatory network for E. coli was integrated with 266 E. coli gene chip observations, POBN identified 93 out of 570 connections that were either inconsistent or not adequately supported by the expression data. CONCLUSION: POBN provides a systematic way to integrate known transcriptional networks with observed gene expression data to better identify which transcriptional pathways are likely responsible for the observed gene expression pattern. PMID- 21619640 TI - Structator: fast index-based search for RNA sequence-structure patterns. AB - BACKGROUND: The secondary structure of RNA molecules is intimately related to their function and often more conserved than the sequence. Hence, the important task of searching databases for RNAs requires to match sequence-structure patterns. Unfortunately, current tools for this task have, in the best case, a running time that is only linear in the size of sequence databases. Furthermore, established index data structures for fast sequence matching, like suffix trees or arrays, cannot benefit from the complementarity constraints introduced by the secondary structure of RNAs. RESULTS: We present a novel method and readily applicable software for time efficient matching of RNA sequence-structure patterns in sequence databases. Our approach is based on affix arrays, a recently introduced index data structure, preprocessed from the target database. Affix arrays support bidirectional pattern search, which is required for efficiently handling the structural constraints of the pattern. Structural patterns like stem loops can be matched inside out, such that the loop region is matched first and then the pairing bases on the boundaries are matched consecutively. This allows to exploit base pairing information for search space reduction and leads to an expected running time that is sublinear in the size of the sequence database. The incorporation of a new chaining approach in the search of RNA sequence-structure patterns enables the description of molecules folding into complex secondary structures with multiple ordered patterns. The chaining approach removes spurious matches from the set of intermediate results, in particular of patterns with little specificity. In benchmark experiments on the Rfam database, our method runs up to two orders of magnitude faster than previous methods. CONCLUSIONS: The presented method's sublinear expected running time makes it well suited for RNA sequence-structure pattern matching in large sequence databases. RNA molecules containing several stem-loop substructures can be described by multiple sequence structure patterns and their matches are efficiently handled by a novel chaining method. Beyond our algorithmic contributions, we provide with Structator a complete and robust open-source software solution for index-based search of RNA sequence-structure patterns. The Structator software is available at http://www.zbh.uni-hamburg.de/Structator. PMID- 21619641 TI - Lack of a-disintegrin-and-metalloproteinase ADAM10 leads to intracellular accumulation and loss of shedding of the cellular prion protein in vivo. AB - BACKGROUND: The cellular prion protein (PrPC) fulfils several yet not completely understood physiological functions. Apart from these functions, it has the ability to misfold into a pathogenic scrapie form (PrPSc) leading to fatal transmissible spongiform encephalopathies. Proteolytic processing of PrPC generates N- and C-terminal fragments which play crucial roles both in the pathophysiology of prion diseases and in transducing physiological functions of PrPC. A-disintegrin-and-metalloproteinase 10 (ADAM10) has been proposed by cell culture experiments to be responsible for both shedding of PrPC and its alpha cleavage. Here, we analyzed the role of ADAM10 in the proteolytic processing of PrPC in vivo. RESULTS: Using neuron-specific Adam10 knockout mice, we show that ADAM10 is the sheddase of PrPC and that its absence in vivo leads to increased amounts and accumulation of PrPC in the early secretory pathway by affecting its posttranslational processing. Elevated PrPC levels do not induce apoptotic signalling via p53. Furthermore, we show that ADAM10 is not responsible for the alpha-cleavage of PrPC. CONCLUSION: Our study elucidates the proteolytic processing of PrPC and proves a role of ADAM10 in shedding of PrPC in vivo. We suggest that ADAM10 is a mediator of PrPC homeostasis at the plasma membrane and, thus, might be a regulator of the multiple functions discussed for PrPC. Furthermore, identification of ADAM10 as the sheddase of PrPC opens the avenue to devising novel approaches for therapeutic interventions against prion diseases. PMID- 21619642 TI - Vaccination coverage and timeliness in three South African areas: a prospective study. AB - BACKGROUND: Timely vaccination is important to induce adequate protective immunity. We measured vaccination timeliness and vaccination coverage in three geographical areas in South Africa. METHODS: This study used vaccination information from a community-based cluster-randomized trial promoting exclusive breastfeeding in three South African sites (Paarl in the Western Cape Province, and Umlazi and Rietvlei in KwaZulu-Natal) between 2006 and 2008. Five interview visits were carried out between birth and up to 2 years of age (median follow-up time 18 months), and 1137 children were included in the analysis. We used Kaplan Meier time-to-event analysis to describe vaccination coverage and timeliness in line with the Expanded Program on Immunization for the first eight vaccines. This included Bacillus Calmette-Guerin (BCG), four oral polio vaccines and 3 doses of the pentavalent vaccine which protects against diphtheria, pertussis, tetanus, hepatitis B and Haemophilus influenzae type B. RESULTS: The proportion receiving all these eight recommended vaccines were 94% in Paarl (95% confidence interval [CI] 91-96), 62% in Rietvlei (95%CI 54-68) and 88% in Umlazi (95%CI 84-91). Slightly fewer children received all vaccines within the recommended time periods. The situation was worst for the last pentavalent- and oral polio vaccines. The hazard ratio for incomplete vaccination was 7.2 (95%CI 4.7-11) for Rietvlei compared to Paarl. CONCLUSIONS: There were large differences between the different South African sites in terms of vaccination coverage and timeliness, with the poorer areas of Rietvlei performing worse than the better-off areas in Paarl. The vaccination coverage was lower for the vaccines given at an older age. There is a need for continued efforts to improve vaccination coverage and timeliness, in particular in rural areas. TRIAL REGISTRATION NUMBER: ClinicalTrials.gov: NCT00397150. PMID- 21619643 TI - An efficient algorithm for systematic analysis of nucleotide strings suitable for siRNA design. AB - BACKGROUND: The "off-target" silencing effect hinders the development of siRNA based therapeutic and research applications. Existing solutions for finding possible locations of siRNA seats within a large database of genes are either too slow, miss a portion of the targets, or are simply not designed to handle a very large number of queries. We propose a new approach that reduces the computational time as compared to existing techniques. FINDINGS: The proposed method employs tree-based storage in a form of a modified truncated suffix tree to sort all possible short string substrings within given set of strings (i.e. transcriptome). Using the new algorithm, we pre-computed a list of the best siRNA locations within each human gene ("siRNA seats"). siRNAs designed to reside within siRNA seats are less likely to hybridize off-target. These siRNA seats could be used as an input for the traditional "set-of-rules" type of siRNA designing software. The list of siRNA seats is available through a publicly available database located at http://web.cos.gmu.edu/~gmanyam/siRNA_db/search.php CONCLUSIONS: In attempt to perform top-down prediction of the human siRNA with minimized off-target hybridization, we developed an efficient algorithm that employs suffix tree based storage of the substrings. Applications of this approach are not limited to optimal siRNA design, but can also be useful for other tasks involving selection of the characteristic strings specific to individual genes. These strings could then be used as siRNA seats, as specific probes for gene expression studies by oligonucleotide-based microarrays, for the design of molecular beacon probes for Real-Time PCR and, generally, any type of PCR primers. PMID- 21619644 TI - Feasibility of inter-hospital transportation using extra-corporeal membrane oxygenation (ECMO) support of patients affected by severe swine-flu(H1N1)-related ARDS. AB - BACKGROUND: To describe the organization of an ECMO-centre from triage by telephone to the phase of inter-hospital transportation with ECMO of patients affected by H1N1-induced ARDS, describing techniques and equipment used. METHODS: From September 2009 to January 2010, 18 patients with H1N1-induced ARDS were referred to our ECMO-centre from other hospitals. Six patients had contraindications to treatment with ECMO and remained in the local hospital. Twelve patients were transported to our centre and were included in this study. Four patients were transported on ECMO (Group A) and eight on conventional ventilation (Group B). The groups were compared on the basis of adverse events during transport, clinical characteristics and outcome. RESULTS: The PaO2/FiO2 ratio was lower in the patients of Group A (46.8 vs 89.7 [median]) despite the PEEP values being higher (15.0 vs 8.5 [median]). The Murray score was higher in Group A (3.50 vs 2.75 [median]). During the transfer there were no significant complications noted in Group A, whereas two patients in Group B were reported with hypoxia (SpO2 < 90%). One patient in Group A died. All the other patients of the two groups have been discharged from hospital. CONCLUSIONS: The creation of an ECMO team, with various experts in the treatment of ARDS, assured a safe transfer of patients with severe hypoxia, over long distances, when in other cases they wouldn't have been be transportable. PMID- 21619645 TI - Still too little qualitative research to shed light on results from reviews of effectiveness trials: a case study of a Cochrane review on the use of lay health workers. AB - BACKGROUND: Qualitative research is used increasingly alongside trials of complex interventions to explore processes, contextual factors, or intervention characteristics that may have influenced trial outcomes. Qualitative research conducted alongside trials can also be used to shed light on the results of systematic reviews of effectiveness by looking for factors that can help explain heterogeneous results across trials. In a Cochrane review on the effects of using lay health workers on maternal and child health and infectious disease control, we identified 82 trials. These trials showed promising benefits but results were heterogeneous. OBJECTIVE: To use qualitative studies conducted alongside these trials to explore factors and processes that might have influenced intervention outcomes. METHODS: We attempted to identify qualitative research carried out alongside the trials by contacting trial authors, checking papers for references to qualitative research, searching Pubmed for related studies, and carrying out citation searches. For those qualitative studies that we included, we extracted information regarding study objective, data collection and analysis methods, and key themes and categories. RESULTS: For 52 (63%) of the trials, we found no qualitative research that had been conducted alongside the trials. For 16 (20%) trials, some form of qualitative data collection had been done but was unavailable or had been done before the trial. For 14 (17%) trials, qualitative research had been done during or shortly after the trial, although descriptions of qualitative methods and results were often sparse. Most of these 14 studies aimed to elicit trial participants' perspectives and experiences of the intervention. A common theme was participants' appreciation of the lay health workers' shared circumstances, for instance with regard to social background or experience of the health condition. In six studies, researchers explored the experiences of the lay health workers themselves. Issues included the importance of regular supervision and health professionals' support or lack of support. CONCLUSIONS: Qualitative studies carried out alongside trials of complex interventions could offer opportunities to authors of systematic reviews of effectiveness wishing to understand the heterogeneity of trial results. For interventions of lay health worker programmes at least, too few such studies exist at present for these opportunities to be realised. PMID- 21619646 TI - Lytic HSV-1 infection induces the multifunctional transcription factor Early Growth Response-1 (EGR-1) in rabbit corneal cells. AB - BACKGROUND: Herpes simplex virus type-1 (HSV-1) infections can cause a number of diseases ranging from simple cold sores to dangerous keratitis and lethal encephalitis. The interaction between virus and host cells, critical for viral replication, is being extensively investigated by many laboratories. In this study, we tested the hypothesis that HSV-1 lytic infection triggers the expression of important multi-functional transcription factor Egr1. The mechanisms of induction are mediated, at least in part, by signaling pathways such as NFkappaB and CREB. METHODS: SIRC, VERO, and 293HEK cell lines were infected with HSV-1, and the Egr-1 transcript and protein were detected by RT-PCR and Western blot, respectively. The localization and expression profile of Egr-1 were investigated further by immunofluorescence microscopy analyses. The recruitment of transcription factors to the Egr-1 promoter during infection was studied by chromatin immunoprecipitation (ChIP). Various inhibitors and dominant negative mutant were used to assess the mechanisms of Egr-1 induction and their effects were addressed by immunofluorescence microscopy. RESULTS: Western blot analyses showed that Egr-1 was absent in uninfected cells; however, the protein was detected 24-72 hours post treatment, and the response was directly proportional to the titer of the virus used for infection. Using recombinant HSV 1 expressing EGFP, Egr-1 was detected only in the infected cells. ChIP assays demonstrated that NFkB and cAMP response element binding protein (CREB) were recruited to the Egr-1 promoter upon infection. Additional studies showed that inhibitors of NFkB and dominant-negative CREB repressed the Egr-1 induction by HSV-1 infection. CONCLUSION: Collectively, these results demonstrate that Egr-1 is expressed rapidly upon HSV-1 infection and that this novel induction could be due to the NFkB/CREB-mediated transactivation. Egr-1 induction might play a key role in the viral gene expression, replication, inflammation, and the disease progression. PMID- 21619647 TI - Reliability of a patient survey assessing cost-related changes in health care use among high deductible health plan enrollees. AB - BACKGROUND: Recent increases in patient cost-sharing for health care have lent increasing importance to monitoring cost-related changes in health care use. Despite the widespread use of survey questions to measure changes in health care use and related behaviors, scant data exists on the reliability of such questions. METHODS: We administered a cross-sectional survey to a stratified random sample of families in a New England health plan's high deductible health plan (HDHP) with >= $500 in annualized out-of-pocket expenditures. Enrollees were asked about their knowledge of their plan, information seeking, behavior change associated with having a deductible, experience of delay in care due in part to cost, and hypothetical delay in care due in part to cost. Initial respondents were mailed a follow-up survey within two weeks of each family returning the original survey. We computed several agreement statistics to measure the test retest reliability for select questions. We also conducted continuity adjusted chi-square, and McNemar tests in both the original and follow-up samples to measure the degree to which our results could be reproduced. Analyses were stratified by self-reported income. RESULTS: The test-retest reliability was moderate for the majority of questions (0.41 - 0.60) and the level of test-retest reliability did not differ substantially across each of the broader domains of questions. The observed proportions of respondents with delayed or foregone pediatric, adult, or any family care were similar when comparing the original and follow-up surveys. In the original survey, respondents in the lower-income group were more likely to delay or forego pediatric care, adult care, or any family care. All of the tests comparing income groups in the follow-up survey produced the same result as in the original survey. CONCLUSIONS: In this population of HDHP beneficiaries, we found that survey questions concerning plan knowledge, information seeking, and delayed or foregone care were moderately reliable. Our results offer reassurance for researchers using survey information to study the effects cost sharing on health care utilization. PMID- 21619648 TI - Study of the IgG endoglycosidase EndoS in group A streptococcal phagocyte resistance and virulence. AB - BACKGROUND: The secreted enzyme EndoS, an endoglycosidase from Streptococcus pyogenes, hydrolyzes the N-linked glycan of the constant region of immunoglobulin G (IgG) heavy chain and renders the antibody unable to interact with Fc receptors and elicit effector functions. In this study we couple targeted allelic replacement mutagenesis and heterologous expression to elucidate the contribution of EndoS to group A Streptococcus (GAS) phagocyte resistance and pathogenicity in vitro and in vivo. RESULTS: Knocking out the EndoS gene in GAS M1T1 background revealed no significant differences in bacterial survival in immune cell killing assays or in a systemic mouse model of infection. However, exogenous addition and heterologous expression of EndoS was found to increase GAS resistance to killing by neutrophils and monocytes in vitro. Additionally, heterologous expression of EndoS in M49 GAS increased mouse virulence in vivo. CONCLUSIONS: We conclude that in a highly virulent M1T1 background, EndoS has no significant impact on GAS phagocyte resistance and pathogenicity. However, local accumulation or high levels of expression of EndoS in certain GAS strains may contribute to virulence. PMID- 21619649 TI - Multi-center feasibility study evaluating recruitment, variability in risk factors and biomarkers for a diet and cancer cohort in India. AB - BACKGROUND: India's population exhibits diverse dietary habits and chronic disease patterns. Nutritional epidemiologic studies in India are primarily of cross-sectional or case-control design and subject to biases, including differential recall of past diet. The aim of this feasibility study was to evaluate whether a diet-focused cohort study of cancer could be established in India, providing insight into potentially unique diet and lifestyle exposures. METHODS: Field staff contacted 7,064 households within three regions of India (New Delhi, Mumbai, and Trivandrum) and found 4,671 eligible adults aged 35-69 years. Participants completed interviewer-administered questionnaires (demographic, diet history, physical activity, medical/reproductive history, tobacco/alcohol use, and occupational history), and staff collected biological samples (blood, urine, and toenail clippings), anthropometric measurements (weight, standing and sitting height; waist, hip, and thigh circumference; triceps, sub-scapula and supra-patella skin fold), and blood pressure measurements. RESULTS: Eighty-eight percent of eligible subjects completed all questionnaires and 67% provided biological samples. Unique protein sources by region were fish in Trivandrum, dairy in New Delhi, and pulses (legumes) in Mumbai. Consumption of meat, alcohol, fast food, and soft drinks was scarce in all three regions. A large percentage of the participants were centrally obese and had elevated blood glucose levels. New Delhi participants were also the least physically active and had elevated lipids levels, suggesting a high prevalence of metabolic syndrome. CONCLUSIONS: A high percentage of participants complied with study procedures including biological sample collection. Epidemiologic expertise and sufficient infrastructure exists at these three sites in India to successfully carry out a modest sized population-based study; however, we identified some potential problems in conducting a cohort study, such as limited number of facilities to handle biological samples. PMID- 21619650 TI - Can molecular cell biology explain chromosome motions? AB - BACKGROUND: Mitotic chromosome motions have recently been correlated with electrostatic forces, but a lingering "molecular cell biology" paradigm persists, proposing binding and release proteins or molecular geometries for force generation. RESULTS: Pole-facing kinetochore plates manifest positive charges and interact with negatively charged microtubule ends providing the motive force for poleward chromosome motions by classical electrostatics. This conceptual scheme explains dynamic tracking/coupling of kinetochores to microtubules and the simultaneous depolymerization of kinetochore microtubules as poleward force is generated. CONCLUSION: We question here why cells would prefer complex molecular mechanisms to move chromosomes when direct electrostatic interactions between known bound charge distributions can accomplish the same task much more simply. PMID- 21619651 TI - Compliant gluten-free children with celiac disease: an evaluation of psychological distress. AB - BACKGROUND: Children with chronic illnesses are known to have increased risks for emotional and behavioral problems. In the present study, children and adolescent suffering from celiac disease (CD) were compared with healthy controls to assess differences in the psychological profile. METHODS: A total of 100 well-treated and compliant CD patients (65 females/35 males; age mean +/- SD: 10.38 +/- 2.71) were compared to 100 normal controls (58 females/42 males; age mean +/- SD: 11.47 +/- 2.61). Emotional and behavioral problems were assessed by the Child Behavior Checklist (CBCL), the Children's Depression Inventory (CDI) and the Multidimensional Anxiety Scale for Children (MASC). RESULTS: Subjects with CD self-reported an increased rate of anxiety and depression symptoms and showed higher scores in "harm avoidance" and "somatic complaints", in the CBCL parent report questionnaire, as compared to healthy control subjects. Furthermore, gender differences could be observed in the group of CD patients, with males displaying significantly higher CBCL externalizing scores, in social, thought and attention problems, as compared to female, who in turns showed more prominent internalizing symptoms such as depression. CONCLUSIONS: The increased rate of emotional and behavioral problems in children and adolescent with CD emphasizes the importance of an early detection of mental health problems in these children. PMID- 21619652 TI - Reduction in BMI z-score and improvement in cardiometabolic risk factors in obese children and adolescents. The Oslo Adiposity Intervention Study - a hospital/public health nurse combined treatment. AB - BACKGROUND: Weight loss and increased physical fitness are established approaches to reduce cardiovascular risk factors. We studied the reduction in BMI z-score associated with improvement in cardiometabolic risk factors in overweight and obese children and adolescents treated with a combined hospital/public health nurse model. We also examined how aerobic fitness influenced the results. METHODS: From 2004-2007, 307 overweight and obese children and adolescents aged 7 17 years were referred to an outpatient hospital pediatrics clinic and evaluated by a multidisciplinary team. Together with family members, they were counseled regarding diet and physical activity at biannual clinic visits. Visits with the public health nurse at local schools or at maternal and child health centres were scheduled between the hospital consultations. Fasting blood samples were taken at baseline and after one year, and aerobic fitness (VO2peak) was measured. In the analyses, 230 subjects completing one year of follow-up by December 2008 were divided into four groups according to changes in BMI z-score: Group 1: decrease in BMI z-score>=0.23, Group 2: decrease in BMI z-score>=0.1-< 0.23, Group 3: decrease in/stable BMI z-score>=0.0-< 0.1, Group 4: increase in BMI z-score (>0.00-0.55). RESULTS: 230 participants were included in the analyses (75%). Mean (SD) BMI z-score was reduced from 2.18 (0.30) to 2.05 (0.39) (p < 0.001) in the group as a whole. After adjustment for BMI z-score, waist circumference and gender, the three groups with reduced BMI z-score had a significantly greater reduction in HOMA-IR, insulin, total cholesterol, LDL cholesterol and total/HDL cholesterol ratio than the group with increased BMI z-score. Adding change in aerobic fitness to the model had little influence on the results. Even a very small reduction in BMI z-score (group 3) was associated with significantly lower insulin, total cholesterol, LDL and total/HDL cholesterol ratio. The group with the largest reduction in BMI z-score had improvements in HOMA-IR and aerobic fitness as well. An increase in BMI z-score was associated with worsening of C peptide and total/HDL cholesterol ratio. CONCLUSIONS: Even a modest reduction in BMI z-score after one year of combined hospital/and public health nurse intervention was associated with improvement in several cardiovascular risk factors. PMID- 21619653 TI - A large, consistent plasma proteomics data set from prospectively collected breast cancer patient and healthy volunteer samples. AB - BACKGROUND: Variability of plasma sample collection and of proteomics technology platforms has been detrimental to generation of large proteomic profile datasets from human biospecimens. METHODS: We carried out a clinical trial-like protocol to standardize collection of plasma from 204 healthy and 216 breast cancer patient volunteers. The breast cancer patients provided follow up samples at 3 month intervals. We generated proteomics profiles from these samples with a stable and reproducible platform for differential proteomics that employs a highly consistent nanofabricated ChipCubeTM chromatography system for peptide detection and quantification with fast, single dimension mass spectrometry (LC MS). Protein identification is achieved with subsequent LC-MS/MS analysis employing the same ChipCubeTM chromatography system. RESULTS: With this consistent platform, over 800 LC-MS plasma proteomic profiles from prospectively collected samples of 420 individuals were obtained. Using a web-based data analysis pipeline for LC-MS profiling data, analyses of all peptide peaks from these plasma LC-MS profiles reveals an average coefficient of variability of less than 15%. Protein identification of peptide peaks of interest has been achieved with subsequent LC-MS/MS analyses and by referring to a spectral library created from about 150 discrete LC-MS/MS runs. Verification of peptide quantity and identity is demonstrated with several Multiple Reaction Monitoring analyses. These plasma proteomic profiles are publicly available through ProteomeCommons. CONCLUSION: From a large prospective cohort of healthy and breast cancer patient volunteers and using a nano-fabricated chromatography system, a consistent LC-MS proteomics dataset has been generated that includes more than 800 discrete human plasma profiles. This large proteomics dataset provides an important resource in support of breast cancer biomarker discovery and validation efforts. PMID- 21619654 TI - In vivo imaging of cell behaviors and F-actin reveals LIM-HD transcription factor regulation of peripheral versus central sensory axon development. AB - BACKGROUND: Development of specific neuronal morphology requires precise control over cell motility processes, including axon formation, outgrowth and branching. Dynamic remodeling of the filamentous actin (F-actin) cytoskeleton is critical for these processes; however, little is known about the mechanisms controlling motile axon behaviors and F-actin dynamics in vivo. Neuronal structure is specified in part by intrinsic transcription factor activity, yet the molecular and cellular steps between transcription and axon behavior are not well understood. Zebrafish Rohon-Beard (RB) sensory neurons have a unique morphology, with central axons that extend in the spinal cord and a peripheral axon that innervates the skin. LIM homeodomain (LIM-HD) transcription factor activity is required for formation of peripheral RB axons. To understand how neuronal morphogenesis is controlled in vivo and how LIM-HD transcription factor activity differentially regulates peripheral versus central axons, we used live imaging of axon behavior and F-actin distribution in vivo. RESULTS: We used an F-actin biosensor containing the actin-binding domain of utrophin to characterize actin rearrangements during specific developmental processes in vivo, including axon initiation, consolidation and branching. We found that peripheral axons initiate from a specific cellular compartment and that F-actin accumulation and protrusive activity precede peripheral axon initiation. Moreover, disruption of LIM-HD transcriptional activity has different effects on the motility of peripheral versus central axons; it inhibits peripheral axon initiation, growth and branching, while increasing the growth rate of central axons. Our imaging revealed that LIM-HD transcription factor activity is not required for F-actin based protrusive activity or F-actin accumulation during peripheral axon initiation, but can affect positioning of F-actin accumulation and axon formation. CONCLUSION: Our ability to image the dynamics of F-actin distribution during neuronal morphogenesis in vivo is unprecedented, and our experiments provide insight into the regulation of cell motility as neurons develop in the intact embryo. We identify specific motile cell behaviors affected by LIM-HD transcription factor activity and reveal how transcription factors differentially control the formation and growth of two axons from the same neuron. PMID- 21619655 TI - LabKey Server NAb: a tool for analyzing, visualizing and sharing results from neutralizing antibody assays. AB - BACKGROUND: Multiple types of assays allow sensitive detection of virus-specific neutralizing antibodies. For example, the extent of antibody neutralization of HIV-1, SIV and SHIV can be measured in the TZM-bl cell line through the degree of luciferase reporter gene expression after infection. In the past, neutralization curves and titers for this standard assay have been calculated using an Excel macro. Updating all instances of such a macro with new techniques can be unwieldy and introduce non-uniformity across multi-lab teams. Using Excel also poses challenges in centrally storing, sharing and associating raw data files and results. RESULTS: We present LabKey Server's NAb tool for organizing, analyzing and securely sharing data, files and results for neutralizing antibody (NAb) assays, including the luciferase-based TZM-bl NAb assay. The customizable tool supports high-throughput experiments and includes a graphical plate template designer, allowing researchers to quickly adapt calculations to new plate layouts. The tool calculates the percent neutralization for each serum dilution based on luminescence measurements, fits a range of neutralization curves to titration results and uses these curves to estimate the neutralizing antibody titers for benchmark dilutions. Results, curve visualizations and raw data files are stored in a database and shared through a secure, web-based interface. NAb results can be integrated with other data sources based on sample identifiers. It is simple to make results public after publication by updating folder security settings. CONCLUSIONS: Standardized tools for analyzing, archiving and sharing assay results can improve the reproducibility, comparability and reliability of results obtained across many labs. LabKey Server and its NAb tool are freely available as open source software at http://www.labkey.com under the Apache 2.0 license. Many members of the HIV research community can also access the LabKey Server NAb tool without installing the software by using the Atlas Science Portal (https://atlas.scharp.org). Atlas is an installation of LabKey Server. PMID- 21619656 TI - A beta-mixture model for dimensionality reduction, sample classification and analysis. AB - BACKGROUND: Patterns of genome-wide methylation vary between tissue types. For example, cancer tissue shows markedly different patterns from those of normal tissue. In this paper we propose a beta-mixture model to describe genome-wide methylation patterns based on probe data from methylation microarrays. The model takes dependencies between neighbour probe pairs into account and assumes three broad categories of methylation, low, medium and high. The model is described by 37 parameters, which reduces the dimensionality of a typical methylation microarray significantly. We used methylation microarray data from 42 colon cancer samples to assess the model. RESULTS: Based on data from colon cancer samples we show that our model captures genome-wide characteristics of methylation patterns. We estimate the parameters of the model and show that they vary between different tissue types. Further, for each methylation probe the posterior probability of a methylation state (low, medium or high) is calculated and the probability that the state is correctly predicted is assessed. We demonstrate that the model can be applied to classify cancer tissue types accurately and that the model provides accessible and easily interpretable data summaries. CONCLUSIONS: We have developed a beta-mixture model for methylation microarray data. The model substantially reduces the dimensionality of the data. It can be used for further analysis, such as sample classification or to detect changes in methylation status between different samples and tissues. PMID- 21619657 TI - The clinical implications of adult-onset henoch-schonelin purpura. AB - Henoch-Schonlein Purpura (HSP) is a small vessel vasculitis mediated by IgA immune complex deposition. It is characterized by the clinical tetrad of non thrombocytopenic palpable purpura, abdominal pain, arthritis and renal involvement. Pathologically, it can be considered a form of immune complex mediated leukocytoclastic vasculitis (LCV) involving the skin and other organs. Though it primarily affects children (over 90% of cases), the occurrence in adults has been rarely reported. Management often involves the use of immunomodulatory or immune-suppressive regimens. PMID- 21619658 TI - Helicobacter pylori with stronger intensity of CagA phosphorylation lead to an increased risk of gastric intestinal metaplasia and cancer. AB - BACKGROUND: Nearly all Taiwanese H. pylori stains are cagA-genopositive and encode CagA protein. In this study, we evaluated whether different intensity of tyrosine phosphorylated-CagA (p-CagA) had an impact on the clinical diseases and histological outcomes in this area. RESULTS: We enrolled 469 dyspeptic patients and prospectively obtained the gastric biopsy specimens and the H. pylori isolates. These patients were categorized according to the clinical diseases, such as duodenal ulcer, gastric ulcer, gastric cancer, and gastritis with or without intestinal metaplasia. Their gastric specimens were reviewed by the updated Sydney's system. Furthermore, a total of 146 patients were randomly selected from each clinical category for evaluation of their isolates' p-CagA intensity by in vitro AGS cells co-culture. The p-CagA was sparse in 30 (20.5%), weak in 59 (40.5%), and strong in 57 (39%) isolates. The isolates from the patients of gastric cancer or gastritis with intestinal metaplasia had stronger p CagA intensity than those of gastritis without intestinal metaplasia (p <= 0.002). Moreover, the patients infected with isolates with strong or weak p-CagA intensity had a higher risk of gastric intestinal metaplasia (p < 0.05, odds ratio 3.09~15.26) than those infected with sparse p-CagA isolates. CONCLUSIONS: Infection with H. pylori stains with stronger p-CagA intensity may lead to an increased risk of gastric intestinal metaplasia and cancer. PMID- 21619659 TI - Synthesis of 86 species of 1,5-diaryl-3-oxo-1,4-pentadienes analogs of curcumin can yield a good lead in vivo. AB - BACKGROUND: Curcumin is known to possess many anti-tumor properties such as inhibition of tumor growth and induction of apotosis. However, limited bioavailability of curcumin prevents its clinical application. A synthesized curcumin analog, 1,5-diaryl-3-oxo-1,4-pentadiene such as GO-Y030, has the improved anti-tumor potential in vitro as well as in mouse model of colorectal carcinogenesis. RESULTS: These compounds were divided into two groups; one is the higher anti-proliferative group, in which 79.7% of 1,5-diaryl-3-oxo-1,4 pentadienes were clustered. One of the 1,5-diaryl-3-oxo-1,4-pentadiene analogs, GO-Y078 has the most enhanced growth inhibition, and its solubility was improved, compared with curcumin. GO-Y078 inhibits NF-kappaB transactivation, as well as expression of TP53 and DR5 more effectively than curcumin. In a mouse model, GO Y078 presented 1.4 fold more survival elongation that was not achieved by curcumin and GO-Y030. CONCLUSIONS: The 1,5-diaryl-3-oxo-1,4-pentadiene analogs can yield good lead compounds for cancer chemotherapy, to overcome low bioavailability of curcumin. PMID- 21619661 TI - Detection of the circulating antigen 14-3-3 protein of Schistosoma japonicum by time-resolved fluoroimmunoassay in rabbits. AB - BACKGROUND: Schistosomiasis remains a major public health concern that afflicts millions of people worldwide. Low levels of Schistosoma infection require more sensitive diagnostic methods. In this study, a time-resolved fluoroimmunoassay (TRFIA) was developed for detecting the signal transduction protein 14-3-3, a circulating antigen of Schistosoma japonicum. RESULTS: The detection limit of 14 3-3-TRFIA was 0.78 ng/ml, with a linear measurement range from 0.78 to 800 ng/ml. The average intra-assay and inter-assay variability of this TRFIA was 8.9% and 12.2% respectively, and the mean recovery rate ranged from 92.1% to 115.5%. Within the first 21 days post-infection in rabbits, the positive rates of the 14 3-3-TRFIA were distinctly higher compared to ELISA. All these findings illustrate that 14-3-3-TRFIA has a higher detection efficacy and is a good early diagnostic method for active Schistosoma infection. CONCLUSIONS: A sandwich TRFIA for detecting the circulating antigen 14-3-3 of S. japonicum has been developed, and has demonstrated to be a good potential diagnostic method for schistosomiasis. PMID- 21619660 TI - Fish-on-a-chip: a sensitive detection microfluidic system for Alzheimer's disease. AB - Microfluidics has become an important tool in diagnosing many diseases, including neurological and genetic disorders. Alzheimer's disease (AD) is a neurodegenerative disease that irreversibly and progressively destroys memory, language ability, and thinking skills. Commonly, detection of AD is expensive and complex. Fluorescence in situ hybridization (FISH)-based microfluidic chip platform is capable of diagnosing AD at an early stage and they are effective tools for the diagnosis with low cost, high speed, and high sensitivity. In this review, we tried to provide basic information on the diagnosis of AD via FISH based microfluidics. Different sample preparations using a microfluidic chip for diagnosis of AD are highlighted. Moreover, rapid innovations in nanotechnology for diagnosis are explained. This review will provide information on dynamic quantification methods for the diagnosis and treatment of AD. The knowledge provided in this review will help develop new integration diagnostic techniques based on FISH and microfluidics. PMID- 21619662 TI - miRFam: an effective automatic miRNA classification method based on n-grams and a multiclass SVM. AB - BACKGROUND: MicroRNAs (miRNAs) are ~22 nt long integral elements responsible for post-transcriptional control of gene expressions. After the identification of thousands of miRNAs, the challenge is now to explore their specific biological functions. To this end, it will be greatly helpful to construct a reasonable organization of these miRNAs according to their homologous relationships. Given an established miRNA family system (e.g. the miRBase family organization), this paper addresses the problem of automatically and accurately classifying newly found miRNAs to their corresponding families by supervised learning techniques. Concretely, we propose an effective method, miRFam, which uses only primary information of pre-miRNAs or mature miRNAs and a multiclass SVM, to automatically classify miRNA genes. RESULTS: An existing miRNA family system prepared by miRBase was downloaded online. We first employed n-grams to extract features from known precursor sequences, and then trained a multiclass SVM classifier to classify new miRNAs (i.e. their families are unknown). Comparing with miRBase's sequence alignment and manual modification, our study shows that the application of machine learning techniques to miRNA family classification is a general and more effective approach. When the testing dataset contains more than 300 families (each of which holds no less than 5 members), the classification accuracy is around 98%. Even with the entire miRBase15 (1056 families and more than 650 of them hold less than 5 samples), the accuracy surprisingly reaches 90%. CONCLUSIONS: Based on experimental results, we argue that miRFam is suitable for application as an automated method of family classification, and it is an important supplementary tool to the existing alignment-based small non-coding RNA (sncRNA) classification methods, since it only requires primary sequence information. AVAILABILITY: The source code of miRFam, written in C++, is freely and publicly available at: http://admis.fudan.edu.cn/projects/miRFam.htm. PMID- 21619663 TI - A prospective study of shoulder pain in primary care: prevalence of imaged pathology and response to guided diagnostic blocks. AB - BACKGROUND: The prevalence of imaged pathology in primary care has received little attention and the relevance of identified pathology to symptoms remains unclear. This paper reports the prevalence of imaged pathology and the association between pathology and response to diagnostic blocks into the subacromial bursa (SAB), acromioclavicular joint (ACJ) and glenohumeral joint (GHJ). METHODS: Consecutive patients with shoulder pain recruited from primary care underwent standardised x-ray, diagnostic ultrasound scan and diagnostic injections of local anaesthetic into the SAB and ACJ. Subjects who reported less than 80% reduction in pain following either of these injections were referred for a magnetic resonance arthrogram (MRA) and GHJ diagnostic block. Differences in proportions of positive and negative imaging findings in the anaesthetic response groups were assessed using Fishers test and odds ratios were calculated a for positive anaesthetic response (PAR) to diagnostic blocks. RESULTS: In the 208 subjects recruited, the rotator cuff and SAB displayed the highest prevalence of pathology on both ultrasound (50% and 31% respectively) and MRA (65% and 76% respectively). The prevalence of PAR following SAB injection was 34% and ACJ injection 14%. Of the 59% reporting a negative anaesthetic response (NAR) for both of these injections, 16% demonstrated a PAR to GHJ injection. A full thickness tear of supraspinatus on ultrasound was associated with PAR to SAB injection (OR 5.02; p < 0.05). Ultrasound evidence of a biceps tendon sheath effusion (OR 8.0; p < 0.01) and an intact rotator cuff (OR 1.3; p < 0.05) were associated with PAR to GHJ injection. No imaging findings were strongly associated with PAR to ACJ injection (p <= 0.05). CONCLUSIONS: Rotator cuff and SAB pathology were the most common findings on ultrasound and MRA. Evidence of a full thickness supraspinatus tear was associated with symptoms arising from the subacromial region, and a biceps tendon sheath effusion and an intact rotator cuff were associated with an intra-articular GHJ pain source. When combined with clinical information, these results may help guide diagnostic decision making in primary care. PMID- 21619664 TI - A gossypiboma (foreign body granuloma) mimicking a residual odontogenic cyst in the mandible: a case report. AB - INTRODUCTION: Gossypiboma (foreign body granuloma) in the tooth socket as a complication of tooth removal is rare. Several cases of gossypiboma have been reported after orthopedic, abdominal, otorhinolaryngology, or plastic surgery, but there has been only one reported case after oral surgery. CASE PRESENTATION: A 42-year-old Caucasian German-speaking Swiss woman applied to our clinic for removal of her right mandibular first molar. Her right mandibular third molar had been removed seven years ago. Post-operatively, she complained of pain and foreign body sensation for six months in the area of the removed tooth. A panoramic radiograph of our patient showed a defined and oval radiolucent area in the socket of the right mandibular third molar evoking a residual cyst. An operation was planned to remove the cyst-like lesion. During surgery, a foreign body composed of gauze was found in the right mandibular third molar region. The histological findings were compatible with a foreign body reaction around gauze. CONCLUSION: Retained gauze must be considered if patients complain of pain and foreign body sensation after tooth removal. The use of gauze with radio-opaque markers and extensive irrigation of the socket with saline to remove gauze fragments can avoid this mishap. PMID- 21619665 TI - Traditional electrosurgery and a low thermal injury dissection device yield different outcomes following bilateral skin-sparing mastectomy: a case report. AB - INTRODUCTION: Although a skin- and nipple-sparing mastectomy technique offers distinct cosmetic and reconstructive advantages over traditional methods, partial skin flap and nipple necrosis remain a significant source of post-operative morbidity. Prior work has suggested that collateral thermal damage resulting from electrocautery use during skin flap development is a potential source of this complication. This report describes the case of a smoker with recurrent ductal carcinoma in situ (DCIS) who experienced significant unilateral skin necrosis following bilateral skin-sparing mastectomy while participating in a clinical trial examining mastectomy outcomes with two different surgical devices. This unexpected complication has implications for the choice of dissection devices in procedures requiring skin flap preservation. CASE PRESENTATION: The patient was a 61-year-old Caucasian woman who was a smoker with recurrent DCIS of her right breast. As part of the clinical trial, each breast was randomized to either the standard of care treatment group (a scalpel and a traditional electrosurgical device) or treatment with a novel, low thermal injury dissection device, allowing for a direct, internally controlled comparison of surgical outcomes. Post operative follow-up at six days was unremarkable for both operative sites. At 16 days post-surgery, the patient presented with a significant wound necrosis in the mastectomy site randomized to the control study group. Following debridement and closure, this site progressively healed over 10 weeks. The contralateral mastectomy, randomized to the alternative device, healed normally. CONCLUSION: We hypothesize that thermal damage to the subcutaneous microvasculature during flap dissection may have contributed to this complication and that the use of a low thermal injury dissection device may be advantageous in select patients undergoing skin- and nipple-sparing mastectomy. PMID- 21619666 TI - Primary central nervous system lymphoma presenting as a pure third ventricular lesion: a case report. AB - INTRODUCTION: Primary central nervous system lymphomas are infrequently occurring lymphomas that account for only 0.3-1.5% of all intra-cranial neoplasms in patients without acquired immune deficiency syndrome. However, a pure third ventricle lymphoma is extremely rare. Here, we discuss the similar radiological appearances of lesions localized in the third ventricle and the importance of accurately diagnosing primary central nervous system lymphomas for favorable treatment outcomes. CASE PRESENTATION: A 38-year-old Caucasian man from Turkey presented with a severe headache lasting for three months that failed to respond to any medication. Both severity and duration of the symptoms increased gradually, resulting in vomiting, nausea and gait disturbance that accompanied the headache for three weeks. Neuro-imaging studies showed a lesion located solely in the third ventricle, resulting in partial obstruction of the foramen of Monro. The pre-operative diagnosis was a colloid cyst. Following the surgical procedure, the results of pathological and immunochemical assays revealed that the pre-operative diagnosis was incorrect and that the lesion was a primary central system lymphoma. CONCLUSION: Pure third ventricle lymphomas are extremely rare and are exceptionally localized. It is important to be aware of, and to differentiate between, other possible third ventricular lesions that may mimic the same radiological appearance. Accurate diagnosis is necessary for selecting appropriate treatment modalities. PMID- 21619667 TI - Retroperitoneal smooth muscle tumor of uncertain malignant potential after hysterectomy: a case report. AB - INTRODUCTION: Smooth muscle tumors of uncertain malignant potential represent a histologically heterogeneous group of uterine smooth muscle tumors that cannot be diagnosed as either benign or malignant. Smooth muscle tumors of uncertain malignant potential are usually clinically benign, but should be considered tumors of low malignant potential because they can occasionally recur or metastasize to distant sites. CASE PRESENTATION: We report the case of a 62-year old Mongol woman diagnosed with a retroperitoneal smooth muscle tumor of uncertain malignant potential and lung metastasis, with a history of prior hysterectomy. The case was initially misdiagnosed as retroperitoneal sarcoma, and our patient received chemotherapy. However, no interval change in the size of the retroperitoneal mass and metastatic lung nodules was seen over a period of at least five years. She underwent partial resection of the retroperitoneal mass for the purposes of debulking and establishing a histopathological diagnosis. The diagnosis of the retroperitoneal mass was then confirmed as a smooth muscle tumor of uncertain malignant potential. CONCLUSION: Smooth muscle tumors of uncertain malignant potential have an unpredictable clinical course, and relapses generally appear to occur after a long disease-free interval of up to several years. Therefore, patients diagnosed with smooth muscle tumors of uncertain malignant potential should receive long-term follow-up. PMID- 21619668 TI - The predictive value of ICD-10 diagnostic coding used to assess Charlson comorbidity index conditions in the population-based Danish National Registry of Patients. AB - BACKGROUND: The Charlson comorbidity index is often used to control for confounding in research based on medical databases. There are few studies of the accuracy of the codes obtained from these databases. We examined the positive predictive value (PPV) of the ICD-10 diagnostic coding in the Danish National Registry of Patients (NRP) for the 19 Charlson conditions. METHODS: Among all hospitalizations in Northern Denmark between 1 January 1998 and 31 December 2007 with a first-listed diagnosis of a Charlson condition in the NRP, we selected 50 hospital contacts for each condition. We reviewed discharge summaries and medical records to verify the NRP diagnoses, and computed the PPV as the proportion of confirmed diagnoses. RESULTS: A total of 950 records were reviewed. The overall PPV for the 19 Charlson conditions was 98.0% (95% CI; 96.9, 98.8). The PPVs ranged from 82.0% (95% CI; 68.6%, 91.4%) for diabetes with diabetic complications to 100% (one-sided 97.5% CI; 92.9%, 100%) for congestive heart failure, peripheral vascular disease, chronic pulmonary disease, mild and severe liver disease, hemiplegia, renal disease, leukaemia, lymphoma, metastatic tumour, and AIDS. CONCLUSION: The PPV of NRP coding of the Charlson conditions was consistently high. PMID- 21619669 TI - Immunological abnormalities as potential biomarkers in Chronic Fatigue Syndrome/Myalgic Encephalomyelitis. AB - BACKGROUND: Chronic Fatigue Syndrome/Myalgic Encephalomyelitis (CFS/ME) is characterised by severe prolonged fatigue, and decreases in cognition and other physiological functions, resulting in severe loss of quality of life, difficult clinical management and high costs to the health care system. To date there is no proven pathomechanism to satisfactorily explain this disorder. Studies have identified abnormalities in immune function but these data are inconsistent. We investigated the profile of markers of immune function (including novel markers) in CFS/ME patients. METHODS: We included 95 CFS/ME patients and 50 healthy controls. All participants were assessed on natural killer (NK) and CD8(+) T cell cytotoxic activities, Th1 and Th2 cytokine profile of CD4(+) T cells, expression of vasoactive intestinal peptide receptor 2 (VPACR2), levels of NK phenotypes (CD56(bright) and CD56(dim)) and regulatory T cells expressing FoxP3 transcription factor. RESULTS: Compared to healthy individuals, CFS/ME patients displayed significant increases in IL-10, IFN-gamma, TNF-alpha, CD4(+)CD25(+) T cells, FoxP3 and VPACR2 expression. Cytotoxic activity of NK and CD8(+) T cells and NK phenotypes, in particular the CD56(bright) NK cells were significantly decreased in CFS/ME patients. Additionally granzyme A and granzyme K expression were reduced while expression levels of perforin were significantly increased in the CFS/ME population relative to the control population. These data suggest significant dysregulation of the immune system in CFS/ME patients. CONCLUSIONS: Our study found immunological abnormalities which may serve as biomarkers in CFS/ME patients with potential for an application as a diagnostic tool. PMID- 21619670 TI - Glycemic indices of five varieties of dates in healthy and diabetic subjects. AB - BACKGROUND: This study was designed to determine the glycemic indices of five commonly used varieties of dates in healthy subjects and their effects on postprandial glucose excursions in individuals with type 2 diabetes mellitus. METHODS: Composition analysis was carried out for five types of dates (Tamer stage). The weights of the flesh of the dates equivalent to 50 g of available carbohydrates were calculated. The study subjects were thirteen healthy volunteers with a mean (+/- SD) age of 40.2 +/- 6.7 years and ten participants with type 2 diabetes mellitus (controlled on lifestyle measures and/or metformin) with a mean HbA1c (+/- SD) of 6.6 +/- (0.7%) and a mean age (+/- SD) of 40.8 +/- 5.7 years. Each subject was tested on eight separate days with 50 g of glucose (on 3 occasions) and 50 g equivalent of available carbohydrates from the 5 varieties of date (each on one occasion). Capillary glucose was measured in the healthy subjects at 0, 15, 30, 45, 60, 90 and 120 min and for the diabetics at 0, 30, 60, 90, 120, 150 and 180 min. The glycemic indices were determined as ratios of the incremental areas under the response curves for the dates compared to glucose. Statistical analyses were performed using the Mann-Whitney U test and repeated measures analysis of variance. RESULTS: Mean glycemic indices +/- SEM of the dates for the healthy individuals were 54.0 +/- 6.1, 53.5 +/- 8.6, 46.3 +/- 7.1, 49.1 +/- 3.6 and 55.1 +/- 7.7 for Fara'd, Lulu, Bo ma'an, Dabbas and Khalas, respectively. Corresponding values for those with type 2 diabetes were very similar (46.1 +/- 6.2, 43.8 +/- 7.7, 51.8 +/- 6.9, 50.2 +/- 3.9 and 53.0 +/- 6.0). There were no statistically significant differences in the GIs between the control and the diabetic groups for the five types of dates, nor were there statistically significant differences among the dates' GIs (df = 4, F = 0.365, p = 0.83). CONCLUSION: The results show low glycemic indices for the five types of dates included in the study and that their consumption by diabetic individuals does not result in significant postprandial glucose excursions. These findings point to the potential benefits of dates for diabetic subjects when used in a healthy balanced diet. TRIAL REGISTRATION NUMBER: ClinicalTrials.gov NCT01307904. PMID- 21619671 TI - Minichromosome Maintenance Protein 7 is a potential therapeutic target in human cancer and a novel prognostic marker of non-small cell lung cancer. AB - BACKGROUND: The research emphasis in anti-cancer drug discovery has always been to search for a drug with the greatest antitumor potential but fewest side effects. This can only be achieved if the drug used is against a specific target located in the tumor cells. In this study, we evaluated Minichromosome Maintenance Protein 7 (MCM7) as a novel therapeutic target in cancer. RESULTS: Immunohistochemical analysis showed that MCM7 was positively stained in 196 of 331 non-small cell lung cancer (NSCLC), 21 of 29 bladder tumor and 25 of 70 liver tumor cases whereas no significant staining was observed in various normal tissues. We also found an elevated expression of MCM7 to be associated with poor prognosis for patients with NSCLC (P = 0.0055). qRT-PCR revealed a higher expression of MCM7 in clinical bladder cancer tissues than in corresponding non neoplastic tissues (P < 0.0001), and we confirmed that a wide range of cancers also overexpressed MCM7 by cDNA microarray analysis. Suppression of MCM7 using specific siRNAs inhibited incorporation of BrdU in lung and bladder cancer cells overexpressing MCM7, and suppressed the growth of those cells more efficiently than that of normal cell strains expressing lower levels of MCM7. CONCLUSIONS: Since MCM7 expression was generally low in a number of normal tissues we examined, MCM7 has the characteristics of an ideal candidate for molecular targeted cancer therapy in various tumors and also as a good prognostic biomarker for NSCLC patients. PMID- 21619672 TI - Self-perceived competence correlates poorly with objectively measured competence in evidence based medicine among medical students. AB - BACKGROUND: Previous studies report various degrees of agreement between self perceived competence and objectively measured competence in medical students. There is still a paucity of evidence on how the two correlate in the field of Evidence Based Medicine (EBM). We undertook a cross-sectional study to evaluate the self-perceived competence in EBM of senior medical students in Malaysia, and assessed its correlation to their objectively measured competence in EBM. METHODS: We recruited a group of medical students in their final six months of training between March and August 2006. The students were receiving a clinically integrated EBM training program within their curriculum. We evaluated the students' self-perceived competence in two EBM domains ("searching for evidence" and "appraising the evidence") by piloting a questionnaire containing 16 relevant items, and objectively assessed their competence in EBM using an adapted version of the Fresno test, a validated tool. We correlated the matching components between our questionnaire and the Fresno test using Pearson's product-moment correlation. RESULTS: Forty-five out of 72 students in the cohort (62.5%) participated by completing the questionnaire and the adapted Fresno test concurrently. In general, our students perceived themselves as moderately competent in most items of the questionnaire. They rated themselves on average 6.34 out of 10 (63.4%) in "searching" and 44.41 out of 57 (77.9%) in "appraising". They scored on average 26.15 out of 60 (43.6%) in the "searching" domain and 57.02 out of 116 (49.2%) in the "appraising" domain in the Fresno test. The correlations between the students' self-rating and their performance in the Fresno test were poor in both the "searching" domain (r = 0.13, p = 0.4) and the "appraising" domain (r = 0.24, p = 0.1). CONCLUSIONS: This study provides supporting evidence that at the undergraduate level, self-perceived competence in EBM, as measured using our questionnaire, does not correlate well with objectively assessed EBM competence measured using the adapted Fresno test. STUDY REGISTRATION: International Medical University, Malaysia, research ID: IMU 110/06. PMID- 21619673 TI - Population pharmacokinetics and pharmacodynamics of hydroxyurea in sickle cell anemia patients, a basis for optimizing the dosing regimen. AB - BACKGROUND: Hydroxyurea (HU) is the first approved pharmacological treatment of sickle cell anemia (SCA). The objectives of this study were to develop population pharmacokinetic(PK)-pharmacodynamic(PD) models for HU in order to characterize the exposure-efficacy relationships and their variability, compare two dosing regimens by simulations and develop some recommendations for monitoring the treatment. METHODS: The models were built using population modelling software NONMEM VII based on data from two clinical studies of SCA adult patients receiving 500-2000 mg of HU once daily. Fetal hemoglobin percentage (HbF%) and mean corpuscular volume (MCV) were used as biomarkers for response. A sequential modelling approach was applied. Models were evaluated using simulation-based techniques. Comparisons of two dosing regimens were performed by simulating 10000 patients in each arm during 12 months. RESULTS: The PK profiles were described by a bicompartmental model. The median (and interindividual coefficient of variation (CV)) of clearance was 11.6 L/h (30%), the central volume was 45.3 L (35%). PK steady-state was reached in about 35 days. For a given dosing regimen, HU exposure varied approximately fivefold among patients. The dynamics of HbF% and MCV were described by turnover models with inhibition of elimination of response. In the studied range of drug exposures, the effect of HU on HbF% was at its maximum (median Imax was 0.57, CV was 27%); the effect on MCV was close to its maximum, with median value of 0.14 and CV of 49%. Simulations showed that 95% of the steady-state levels of HbF% and MCV need 26 months and 3 months to be reached, respectively. The CV of the steady-state value of HbF% was about 7 times larger than that of MCV. Simulations with two different dosing regimens showed that continuous dosing led to a stronger HbF% increase in some patients. CONCLUSIONS: The high variability of response to HU was related in part to pharmacokinetics and to pharmacodynamics. The steady-state value of MCV at month 3 is not predictive of the HbF% value at month 26. Hence, HbF% level may be a better biomarker for monitoring HU treatment. Continuous dosing might be more advantageous in terms of HbF% for patients who have a strong response to HU. TRIAL REGISTRATION: The clinical studies whose data are analysed and reported in this work were not required to be registered in France at their time. Both studies were approved by local ethics committees (of Mondor Hospital and of Kremlin-Bicetre Hospital) and written informed consent was obtained from each patient. PMID- 21619674 TI - Impact of the introduction of new vaccines and vaccine wastage rate on the cost effectiveness of routine EPI: lessons from a descriptive study in a Cameroonian health district. AB - The Expanded Program of Immunization (EPI) offers services to the population free of charge but these activities are costly with the greatest part being the cost of vaccines. In spite of the growing international solidarity towards funding for immunization, the growing objectives continue to outweigh the available resources. It is therefore crucial for any immunization system to seek greater efficiency so as to optimize the use of available means in a bid to ensure sustainability. It is in this light that we carried out this study which aims to assess the productive efficiency of routine EPI for children aged 0 - 11 months with respect to the fixed and outreach vaccine delivery strategies in Ngong health district. The study is descriptive and cross-sectional. Data were collected retrospectively for all 16 health centers of the district that offered EPI services during the period February - May 2009.The results show that:* Only 62% of planned outreach immunization sessions were effectively carried out mainly due to limited funds for transportation and staff availability. Consequently vaccine coverage was low (BCG: 70.1%, DPT-HB-Hib 3: 55.5%) and less resources (43%) were used for this strategy which served 52% of the target population - a major blow to equity.* The average cost per Fully Immunized Child (FIC) was 9,571 FCFA (19.22 USD) for the fixed strategy; 12,751 FCFA (25.61 USD) for the outreach and 10,718 FCFA (21.53 USD) with both strategies combined. These figures are high than those observed in many other African health districts. However, DPT-HB-Hib and yellow fever vaccines contributed to the increase as vaccines occupied 57% of the total cost. With DPT in lieu of DPT-HB-Hib the cost/FIC would be 6,046 FCFA (12.14 USD). Dropout rates too were high (28.1% for the fixed, 29.7% for outreach).* The cost of vaccines wasted in excess of the national norm at the level of health centers was 595,532 FCFA (1,196.15 USD), an amount that could cover the vaccine cost for 122 FIC (7.6% of the FIC during the period). This was accounted for as follows: BCG 1.1%, OPV 1.4%, DPT-HB-Hib 72.7%, measles 5.3%, yellow fever 19.5%* Therefore we suggest improved communication for EPI, the introduction of DPT-HB-Hib with liquid Hib and the effective implementation of planned outreach sessions. PMID- 21619675 TI - Long-term outcome in relationship to neonatal transfusion volume in extremely premature infants: a comparative cohort study. AB - BACKGROUND: In premature born infants red blood cell (RBC) transfusions have been associated with both beneficial and detrimental sequels. Upon RBC transfusion, improvement in cerebral blood flow and oxygenation have been observed, while a more liberal transfusion policy may be associated with a better developmental outcome. The effect of the transfusion volume on long-term outcome is not known. METHODS: Observational follow-up study of a cohort of extremely premature born infants, treated in 2 neonatal intensive care units using a different transfusion volume (15 ml/kg in Unit A and 20 ml/kg in Unit B). The primary outcome was a composite of post discharge mortality, neuromotor developmental delay, blindness or deafness, evaluated at a mean corrected age (CA) of 24 months related to the transfusion volume/kg bodyweight administered during the postnatal hospital stay. RESULTS: Despite the difference in transfusion volume in clinically comparable groups of infants, they received a similar number of transfusions (5.5 +/- 3.2 versus 5.5 +/- 2.3 respectively in Unit A and B). The total transfused volume in unit A was 79 +/- 47 ml/kg and 108 +/- 47 ml/kg in unit B (p = 0.02). Total transfused RBC volume per kg bodyweight was not an independent predictor of the composite outcome (p = 0.96, OR 1.0 (CI 0.9-1.1). CONCLUSION: There was no relationship between the composite outcome at 24 months CA and transfusion volume received during the post natal hospital stay. As there was no clinical advantage of the higher transfusion volume, a more restrictive volume will reduce total transfusion volume and donor exposure. Future research on the optimal transfusion volume per event to extreme preterm infants should include larger, prospective studies with a longer follow-up period through to childhood or even adolescence. PMID- 21619676 TI - Non-invasive evaluation of myocardial reperfusion by transthoracic Doppler echocardiography and single-photon emission computed tomography in patients with anterior acute myocardial infarction. AB - BACKGROUND: The study was designed to evaluate whether the preserved coronary flow reserve (CFR) 72 hours after reperfused acute myocardial infarction (AMI) is associated with less microvascular dysfunction and is predictive of left ventricular (LV) functional recovery and the final infarct size at follow-up. METHODS: In our study, CFR was assessed by transthoracic Doppler echocardiography (TDE) in 44 patients after the successful percutaneous coronary intervention during the acute AMI phase. CFR was correlated with contractile reserve assessed by low-dose dobutamine echocardiography and with the total perfusion defect measured by single-photon emission computed tomography 72 hours after reperfusion and at 5 months follow-up. The ROC analysis was performed to determine test sensitivity and specificity based on CFR. Categorical data were compared by an chi2 analysis, continuous variables were analysed with the independent Student's t test. In order to analyse correlation between CFR and the parameters of LV function and perfusion, the Pearson correlation analysis was conducted. The linear regression analysis was used to assess the relationship between CFR and myocardial contractility as well as the final infarct size. RESULTS: We estimated the CFR cut-off value of 1.75 as providing the maximal accuracy to distinguish between patients with preserved and impaired CFR during the acute AMI phase (sensitivity 91.7%, specificity 75%). Wall motion score index was better in the subgroup with preserved CFR as compared to the subgroup with reduced CFR: 1.74 (0.29) vs. 1.89 (0.17) (p < 0.001) during the acute phase and 1.47 (0.30) vs. 1.81 (0.20) (p < 0.001) at follow-up, respectively. LV ejection fraction was 47.78% (8.99) in preserved CFR group vs. 40.79% (7.25) in impaired CFR group (p = 0.007) 72 hours after reperfusion and 49.78% (8.70) vs. 40.36% (7.90) (p = 0.001) after 5 months at follow-up, respectively. The final infarct size was smaller in patients with preserved as compared to patients with reduced CFR: 5.26% (6.14) vs. 23.28% (12.19) (p < 0.001) at follow-up. CONCLUSION: The early measurement of CFR by TDE can be of high value for the assessment of successful reperfusion in AMI and can be used to predict LV functional recovery, myocardial viability and the final infarct size. PMID- 21619677 TI - Seatbelts and road traffic collision injuries. AB - Modification of seatbelts and their legislation played an important role in reducing morbidity and mortality of occupants in road traffic collisions. We aimed to review seatbelt development, its mechanism of action and its effects. Seatbelts reduce injury by preventing the occupant from hitting the interior parts of the vehicle or being ejected from the car. We have made a linear regression correlation between the overall seatbelt compliance and road traffic death rates in 46 high income countries to study the relationship between seatbelt use and mortality. There was a very highly significant negative correlation between the seatbelt compliance and road traffic death rates (R = - 0.77, F = 65.5, p < 0.00001). Seatbelt-related injuries include spinal, abdominal or pelvic injuries. The presence of a seatbelt sign must raise the suspicion of an intra-abdominal injury. These injuries can be reduced if seatbelts were applied correctly. Although seatbelts were recognized as an important safety measure, it still remains underused in many countries. Enforcement of seatbelt usage by law is mandatory so as to reduce the toll of death of road traffic collisions. PMID- 21619678 TI - A predicted protein, KIAA0247, is a cell cycle modulator in colorectal cancer cells under 5-FU treatment. AB - BACKGROUND: Colorectal cancer (CRC) is the predominant gastrointestinal malignancy and the leading cause of cancer death. The identification of genes related to CRC is important for the development of successful therapies and earlier diagnosis. METHODS: Molecular analysis of feces was evaluated as a potential method for CRC detection. Expression of a predicted protein with unknown function, KIAA0247, was found in feces evaluated using specific quantitative real-time polymerase chain reaction. Its cellular function was then analyzed using immunofluorescent staining and the changes in the cell cycle in response to 5-fluorouracil (5-FU) were assessed. RESULTS: Gastrointestinal tissues and peripheral blood lymphocytes ubiquitously expressed KIAA0247. 56 CRC patients fell into two group categories according to fecal KIAA0247 mRNA expression levels. The group with higher fecal KIAA0247 (n=22; >=0.4897) had a significantly greater five-year overall survival rate than the group with lower fecal KIAA0247 (n = 30; <0.4897) (66.0 +/- 11.6%; p=0.035, log-rank test). Fecal expression of KIAA0247 inversely related to CRC tumor size (Kendall's tau-b= 0.202; p=0.047). Immunofluorescent staining revealed that the cytoplasm of CRC cells evenly expresses KIAA0247 without 5-FU treatment, and KIAA0247 accumulates in the nucleus after 40 MUM 5-FU treatment. In HCT116 p53(-/-) cells, which lack p53 cell cycle control, the proportion of cells in the G2/M phase was larger (13%) in KIAA0247-silent cells than in the respective shLuc control (10%) and KIAA0247-overexpressing cells (7%) after the addition of low dose (40 MUM) 5-FU. Expression of three cyclin genes (cyclin A2, cyclin B1, and cyclin B2) also downregulated in the cells overexpressing KIAA0247. CONCLUSIONS: This is the first description of a linkage between KIAA0247 and CRC. The study's data demonstrate overexpression of KIAA0247 associates with 5-FU therapeutic benefits, and also identify the clinical significance of fecal KIAA0247 in CRC. PMID- 21619679 TI - The evolutionary history of the SAL1 gene family in eutherian mammals. AB - BACKGROUND: SAL1 (salivary lipocalin) is a member of the OBP (Odorant Binding Protein) family and is involved in chemical sexual communication in pig. SAL1 and its relatives may be involved in pheromone and olfactory receptor binding and in pre-mating behaviour. The evolutionary history and the selective pressures acting on SAL1 and its orthologous genes have not yet been exhaustively described. The aim of the present work was to study the evolution of these genes, to elucidate the role of selective pressures in their evolution and the consequences for their functions. RESULTS: Here, we present the evolutionary history of SAL1 gene and its orthologous genes in mammals. We found that (1) SAL1 and its related genes arose in eutherian mammals with lineage-specific duplications in rodents, horse and cow and are lost in human, mouse lemur, bushbaby and orangutan, (2) the evolution of duplicated genes of horse, rat, mouse and guinea pig is driven by concerted evolution with extensive gene conversion events in mouse and guinea pig and by positive selection mainly acting on paralogous genes in horse and guinea pig, (3) positive selection was detected for amino acids involved in pheromone binding and amino acids putatively involved in olfactory receptor binding, (4) positive selection was also found for lineage, indicating a species-specific strategy for amino acid selection. CONCLUSIONS: This work provides new insights into the evolutionary history of SAL1 and its orthologs. On one hand, some genes are subject to concerted evolution and to an increase in dosage, suggesting the need for homogeneity of sequence and function in certain species. On the other hand, positive selection plays a role in the diversification of the functions of the family and in lineage, suggesting adaptive evolution, with possible consequences for speciation and for the reinforcement of prezygotic barriers. PMID- 21619680 TI - Molecular evolution of the vertebrate TLR1 gene family--a complex history of gene duplication, gene conversion, positive selection and co-evolution. AB - BACKGROUND: The Toll-like receptors represent a large superfamily of type I transmembrane glycoproteins, some common to a wide range of species and others are more restricted in their distribution. Most members of the Toll-like receptor superfamily have few paralogues; the exception is the TLR1 gene family with four closely related genes in mammals TLR1, TLR2, TLR6 and TLR10, and four in birds TLR1A, TLR1B, TLR2A and TLR2B. These genes were previously thought to have arisen by a series of independent gene duplications. To understand the evolutionary pattern of the TLR1 gene family in vertebrates further, we cloned the sequences of TLR1A, TLR1B, TLR2A and TLR2B in duck and turkey, constructed phylogenetic trees, predicted codons under positive selection and identified co-evolutionary amino acid pairs within the TLR1 gene family using sequences from 4 birds, 28 mammals, an amphibian and a fish. RESULTS: This detailed phylogenetic analysis not only clarifies the gene gains and losses within the TLR1 gene family of birds and mammals, but also defines orthologues between these vertebrates. In mammals, we predict amino acid sites under positive selection in TLR1, TLR2 and TLR6 but not TLR10. We detect co-evolution between amino acid residues in TLR2 and the other members of this gene family predicted to maintain their ability to form functional heterodimers. In birds, we predict positive selection in the TLR2A and TLR2B genes at functionally significant amino acid residues. We demonstrate that the TLR1 gene family has mostly been subject to purifying selection but has also responded to directional selection at a few sites, possibly in response to pathogen challenge. CONCLUSIONS: Our phylogenetic and structural analyses of the vertebrate TLR1 family have clarified their evolutionary origins and predict amino acid residues likely to be important in the host's defense against invading pathogens. PMID- 21619681 TI - Pulsatile lavage irrigator tip, a rare radiolucent retained foreign body in the pelvis: a case report. AB - Retained foreign bodies after surgery have the potential to cause serious medical complications for patients and bring fourth serious medico-legal consequences for surgeons and hospitals. Standard operating room protocols have been adopted to reduce the occurrence of the most common retained foreign bodies. Despite these precautions, radiolucent objects and uncounted components/pieces of instruments are at risk to be retained in the surgical wound. We report the unusual case of a retained plastic pulsatile lavage irrigator tip in the surgical wound during acetabulum fracture fixation, which was subsequently identified on routine postoperative computed tomography. Revision surgery was required in order to remove the retained object, and the patient had no further complications. PMID- 21619682 TI - Community capacity to acquire, assess, adapt, and apply research evidence: a survey of Ontario's HIV/AIDS sector. AB - BACKGROUND: Community-based organizations (CBOs) are important stakeholders in health systems and are increasingly called upon to use research evidence to inform their advocacy, program planning, and service delivery. To better support CBOs to find and use research evidence, we sought to assess the capacity of CBOs in the HIV/AIDS sector to acquire, assess, adapt, and apply research evidence in their work. METHODS: We invited executive directors of HIV/AIDS CBOs in Ontario, Canada (n = 51) to complete the Canadian Health Services Research Foundation's "Is Research Working for You?" survey. FINDINGS: Based on responses from 25 organizations that collectively provide services to approximately 32,000 clients per year with 290 full-time equivalent staff, we found organizational capacity to acquire, assess, adapt, and apply research evidence to be low. CBO strengths include supporting a culture that rewards flexibility and quality improvement, exchanging information within their organization, and ensuring that their decision-making processes have a place for research. However, CBO Executive Directors indicated that they lacked the skills, time, resources, incentives, and links with experts to acquire research, assess its quality and reliability, and summarize it in a user-friendly way. CONCLUSION: Given the limited capacity to find and use research evidence, we recommend a capacity-building strategy for HIV/AIDS CBOs that focuses on providing the tools, resources, and skills needed to more consistently acquire, assess, adapt, and apply research evidence. Such a strategy may be appropriate in other sectors and jurisdictions as well given that CBO Executive Directors in the HIV/AIDS sector in Ontario report low capacity despite being in the enviable position of having stable government infrastructure in place to support them, benefiting from long-standing investment in capacity building, and being part of an active provincial network. CBOs in other sectors and jurisdictions that have fewer supports may have comparable or lower capacity. Future research should examine a larger sample of CBO Executive Directors from a range of sectors and jurisdictions. PMID- 21619683 TI - Ribosomal protein S6 kinase (RSK)-2 as a central effector molecule in RON receptor tyrosine kinase mediated epithelial to mesenchymal transition induced by macrophage-stimulating protein. AB - BACKGROUND: Epithelial to mesenchymal transition (EMT) occurs during cancer cell invasion and malignant metastasis. Features of EMT include spindle-like cell morphology, loss of epithelial cellular markers and gain of mesenchymal phenotype. Activation of the RON receptor tyrosine kinase by macrophage stimulating protein (MSP) has been implicated in cellular EMT program; however, the major signaling determinant(s) responsible for MSP-induced EMT is unknown. RESULTS: The study presented here demonstrates that RSK2, a downstream signaling protein of the Ras-Erk1/2 pathway, is the principal molecule that links MSP activated RON signaling to complete EMT. Using MDCK cells expressing RON as a model, a spindle-shape based screen was conducted, which identifies RSK2 among various intracellular proteins as a potential signaling molecule responsible for MSP-induced EMT. MSP stimulation dissociated RSK2 with Erk1/2 and promoted RSK2 nuclear translocation. MSP strongly induced RSK2 phosphorylation in a dose dependent manner. These effects relied on RON and Erk1/2 phosphorylation, which is significantly potentiated by transforming growth factor (TGF)-beta1, an EMT inducing cytokine. Specific RSK inhibitor SL0101 completely prevented MSP-induced RSK phosphorylation, which results in inhibition of MSP-induced spindle-like morphology and suppression of cell migration associated with EMT. In HT-29 cancer cells that barely express RSK2, forced RSK2 expression results in EMT-like phenotype upon MSP stimulation. Moreover, specific siRNA-mediated silencing of RSK2 but not RSK1 in L3.6pl pancreatic cancer cells significantly inhibited MSP induced EMT-like phenotype and cell migration. CONCLUSIONS: MSP-induced RSK2 activation is a critical determinant linking RON signaling to cellular EMT program. Inhibition of RSK2 activity may provide a therapeutic opportunity for blocking RON-mediated cancer cell migration and subsequent invasion. PMID- 21619684 TI - Development of an ELISA for sensitive and specific detection of IgA autoantibodies against BP180 in pemphigoid diseases. AB - BACKGROUND: Pemphigoids are rare diseases associated with IgG, IgE and IgA autoantibodies against collagen XVII/BP180. An entity of the pemphigoid group is the lamina lucida-type of linear IgA disease (IgA pemphigoid) characterized by IgA autoantibodies against BP180. While for the detection of IgG and IgE autoantibodies specific to collagen XVII several ELISA systems have been established, no quantitative immunoassay has been yet developed for IgA autoantibodies. Therefore, the aim of the present study was to develop an ELISA to detect IgA autoantibodies against collagen XVII in the sera of patients with pemphigoids. METHODS: We expressed a soluble recombinant form of the collagen XVII ectodomain in mammalian cells. Reactivity of IgA autoantibodies from patients with IgA pemphigoid was assessed by immunofluorescence microscopy and immunoblot analysis. ELISA test conditions were determined by chessboard titration experiments. The sensitivity, specificity and the cut-off were determined by receiver-operating characteristics analysis. RESULTS: The optimized assay was carried out using sera from patients with IgA pemphigoid (n = 30) and healthy donors (n=105). By receiver operating characteristics (ROC) analysis, an area under the curve of 0.993 was calculated, indicating an excellent discriminatory capacity. Thus, a sensitivity and specificity of 83.3% and 100%, respectively, was determined for a cut-off point of 0.48. As additional control groups, sera from patients with bullous pemphigoid (n=31) and dermatitis herpetiformis (n = 50), a disease associated with IgA autoantibodies against epidermal transglutaminase, were tested. In 26% of bullous pemphigoid patients, IgA autoantibodies recognized the ectodomain of collagen XVII. One of 50 (2%) of dermatitis herpetiformis patients sera slightly topped the cut-off value. CONCLUSIONS: We developed the first ELISA for the specific and sensitive detection of serum IgA autoantibodies specific to collagen XVII in patients with pemphigoids. This immunoassay should prove a useful tool for clinical and translational research and should essentially improve the diagnosis and disease monitoring of patients with IgA pemphigoid. Moreover, our findings strongly suggest that IgA pemphigoid and IgG bullous pemphigoid represent two ends of the clinical spectrum of an immunological loss of tolerance against components of hemidesmosomes, which is mediated by both IgG and IgA autoantibodies. PMID- 21619685 TI - Lipid lowering by hydroalcoholic extracts of Amaranthus caudatus L. induces regression of rabbits atherosclerotic lesions. AB - BACKGROUND: The antihypercholesterolemic and antiatherogenic effect of hydroalcoholic extracts of Amaranthus caudatus L (A. caudatus). on regression of atherosclerosis in experimental rabbits maintained on a high cholesterol diet. METHODS: Twenty five rabbits were randomly divided into five groups of five each and treated 75 days as follows: Group I: normal diet(ND), Group II: Hypercholesterolemic diet (HCD) for 45 days; Group III: Hypercholesterolemic diet (HCD) for 75 days, Group IV and V: HCD for 45 days and then normal diet and normal diet + A. caudatus(150 mg.kg day) respectively for an additional 30 days(regression period). Blood samples were collected before (0 time) and after 45 days and 75 days of experimental diets for measurement of biochemical factors. The aortas were removed at the end of the study for assessment of atherosclerotic plaques. RESULTS: In regression period dietary use of A. caudatus in group V significantly decreased total cholesterol, LDL-cholesterol, malondialdehyde, C reactive protein while apolipoproteinA and HDL-cholesterol was significantly increased compared to group IV. The atherosclerotic area was significantly decreased in group V. Whereas, the animals that in regression period received only normal diet showed no regression but rather progression of atherosclerosis. CONCLUSION: These results thus suggest that hydroalcoholic extracts of A. caudatus can reduce risk factors and cause regression of fatty lesions in aorta. PMID- 21619686 TI - Small interference RNA targeting tissue factor inhibits human lung adenocarcinoma growth in vitro and in vivo. AB - BACKGROUND: The human coagulation trigger tissue factor (TF) is overexpressed in several types of cancer and involved in tumor growth, vascularization, and metastasis. To explore the role of TF in biological processes of lung adenocarcinoma, we used RNA interference (RNAi) technology to silence TF in a lung adenocarcinoma cell line A549 with high-level expression of TF and evaluate its antitumor effects in vitro and in vivo. METHODS: The specific small interfering RNA (siRNA) designed for targeting human TF was transfected into A549 cells. The expression of TF was detected by reverse transcription-PCR and Western blot. Cell proliferation was measured by MTT and clonogenic assays. Cell apoptosis was assessed by flow cytometry. The metastatic potential of A549 cells was determined by wound healing, the mobility and Matrigel invasion assays. Expressions of PI3K/Akt, Erk1/2, VEGF and MMP-2/-9 in transfected cells were detected by Western blot. In vivo, the effect of TF-siRNA on the growth of A549 lung adenocarcinoma xenografts in nude mice was investigated. RESULTS: TF -siRNA significantly reduced the expression of TF in the mRNA and protein levels. The down-regulation of TF in A549 cells resulted in the suppression of cell proliferation, invasion and metastasis and induced cell apoptosis in dose dependent manner. Erk MAPK, PI3K/Akt pathways as well as VEGF and MMP-2/-9 expressions were inhibited in TF-siRNA transfected cells. Moreover, intratumoral injection of siRNA targeting TF suppressed the tumor growth of A549 cells in vivo model of lung adenocarcinoma. CONCLUSIONS: Down-regulation of TF using siRNA could provide a potential approach for gene therapy against lung adenocarcinoma, and the antitumor effects may be associated with inhibition of Erk MAPK, PI3K/Akt pathways. PMID- 21619687 TI - A combination of transcriptome and methylation analyses reveals embryologically relevant candidate genes in MRKH patients. AB - BACKGROUND: The Mayer-Rokitansky-Kuster-Hauser (MRKH) syndrome is present in at least 1 out of 4,500 female live births and is the second most common cause for primary amenorrhea. It is characterized by vaginal and uterine aplasia in an XX individual with normal secondary characteristics. It has long been considered a sporadic anomaly, but familial clustering occurs. Several candidate genes have been studied although no single factor has yet been identified. Cases of discordant monozygotic twins suggest that the involvement of epigenetic factors is more likely. METHODS: Differences in gene expression and methylation patterns of uterine tissue between eight MRKH patients and eight controls were identified using whole-genome microarray analyses. Results obtained by expression and methylation arrays were confirmed by qRT-PCR and pyrosequencing. RESULTS: We delineated 293 differentially expressed and 194 differentially methylated genes of which nine overlap in both groups. These nine genes are mainly embryologically relevant for the development of the female genital tract. CONCLUSION: Our study used, for the first time, a combined whole-genome expression and methylation approach to reveal the etiology of the MRKH syndrome. The findings suggest that either deficient estrogen receptors or the ectopic expression of certain HOXA genes might lead to abnormal development of the female reproductive tract. In utero exposure to endocrine disruptors or abnormally high maternal hormone levels might cause ectopic expression or anterior transformation of HOXA genes. It is, however, also possible that different factors influence the anti-Mullerian hormone promoter activity during embryological development causing regression of the Mullerian ducts. Thus, our data stimulate new research directions to decipher the pathogenic basis of MRKH syndrome. PMID- 21619688 TI - Breast metastasis of primary colon cancer with micrometastasis in the axillary sentinel node: a metastasis that metastasized? AB - A case of single breast metastasis from colon adenocarcinoma, with omolateral axillary micrometastasis, is reported with a brief review of the pertinent literature. The originality of the oncological concept of metastasis from metastasis, through lymphatics penetration, is discussed in the setting of a rare condition of breast metastasis from a colorectal carcinoma. The demonstration of axillary lymph node micrometastasis has been possible because fine needle aspiration cytology of the breast nodule was suspicious, but not conclusive for metastasis from colon cancer, so lumpectomy with sentinel node biopsy was planned. Although no disseminated nodal metastases were evident on computerized tomography scan and ultrasonography before breast surgery, the patient developed brain metastases and deteriorated rapidly; she died 16 months after presenting with the breast mass. In conclusion, solid cancers are able to further metastasize, via well-known pathways also recognized in primary cancers such as neoplastic cell invasion of peritumoral lymphatics. PMID- 21619689 TI - Applying psychological theories to evidence-based clinical practice: identifying factors predictive of lumbar spine x-ray for low back pain in UK primary care practice. AB - BACKGROUND: Psychological models predict behaviour in a wide range of settings. The aim of this study was to explore the usefulness of a range of psychological models to predict the health professional behaviour 'referral for lumbar spine x ray in patients presenting with low back pain' by UK primary care physicians. METHODS: Psychological measures were collected by postal questionnaire survey from a random sample of primary care physicians in Scotland and north England. The outcome measures were clinical behaviour (referral rates for lumbar spine x rays), behavioural simulation (lumbar spine x-ray referral decisions based upon scenarios), and behavioural intention (general intention to refer for lumbar spine x-rays in patients with low back pain). Explanatory variables were the constructs within the Theory of Planned Behaviour (TPB), Social Cognitive Theory (SCT), Common Sense Self-Regulation Model (CS-SRM), Operant Learning Theory (OLT), Implementation Intention (II), Weinstein's Stage Model termed the Precaution Adoption Process (PAP), and knowledge. For each of the outcome measures, a generalised linear model was used to examine the predictive value of each theory individually. Linear regression was used for the intention and simulation outcomes, and negative binomial regression was used for the behaviour outcome. Following this 'theory level' analysis, a 'cross-theoretical construct' analysis was conducted to investigate the combined predictive value of all individual constructs across theories. RESULTS: Constructs from TPB, SCT, CS-SRM, and OLT predicted behaviour; however, the theoretical models did not fit the data well. When predicting behavioural simulation, the proportion of variance explained by individual theories was TPB 11.6%, SCT 12.1%, OLT 8.1%, and II 1.5% of the variance, and in the cross-theory analysis constructs from TPB, CS-SRM and II explained 16.5% of the variance in simulated behaviours. When predicting intention, the proportion of variance explained by individual theories was TPB 25.0%, SCT 21.5%, CS-SRM 11.3%, OLT 26.3%, PAP 2.6%, and knowledge 2.3%, and in the cross-theory analysis constructs from TPB, SCT, CS-SRM, and OLT explained 33.5% variance in intention. Together these results suggest that physicians' beliefs about consequences and beliefs about capabilities are likely determinants of lumbar spine x-ray referrals. CONCLUSIONS: The study provides evidence that taking a theory-based approach enables the creation of a replicable methodology for identifying factors that predict clinical behaviour. However, a number of conceptual and methodological challenges remain. PMID- 21619690 TI - Evaluation of lymph node numbers for adequate staging of Stage II and III colon cancer. AB - BACKGROUND: Although evaluation of at least 12 lymph nodes (LNs) is recommended as the minimum number of nodes required for accurate staging of colon cancer patients, there is disagreement on what constitutes an adequate identification of such LNs. METHODS: To evaluate the minimum number of LNs for adequate staging of Stage II and III colon cancer, 490 patients were categorized into groups based on 1-6, 7-11, 12-19, and >= 20 LNs collected. RESULTS: For patients with Stage II or III disease, examination of 12 LNs was not significantly associated with recurrence or mortality. For Stage II (HR = 0.33; 95% CI, 0.12-0.91), but not for Stage III patients (HR = 1.59; 95% CI, 0.54-4.64), examination of >=20 LNs was associated with a reduced risk of recurrence within 2 years. However, examination of >=20 LNs had a 55% (Stage II, HR = 0.45; 95% CI, 0.23-0.87) and a 31% (Stage III, HR = 0.69; 95% CI, 0.38-1.26) decreased risk of mortality, respectively. For each six additional LNs examined from Stage III patients, there was a 19% increased probability of finding a positive LN (parameter estimate = 0.18510, p < 0.0001). For Stage II and III colon cancers, there was improved survival and a decreased risk of recurrence with an increased number of LNs examined, regardless of the cutoff-points. Examination of >=7 or >=12 LNs had similar outcomes, but there were significant outcome benefits at the >=20 cutoff-point only for Stage II patients. For Stage III patients, examination of 6 additional LNs detected one additional positive LN. CONCLUSIONS: Thus, the 12 LN cut-off point cannot be supported as requisite in determining adequate staging of colon cancer based on current data. However, a minimum of 6 LNs should be examined for adequate staging of Stage II and III colon cancer patients. PMID- 21619692 TI - Brazil nuts intake improves lipid profile, oxidative stress and microvascular function in obese adolescents: a randomized controlled trial. AB - BACKGROUND: Obesity is a chronic disease associated to an inflammatory process resulting in oxidative stress that leads to morpho-functional microvascular damage that could be improved by some dietary interventions. In this study, the intake of Brazil nuts (Bertholletia excelsa), composed of bioactive substances like selenium, alpha- e gamma- tocopherol, folate and polyunsaturated fatty acids, have been investigated on antioxidant capacity, lipid and metabolic profiles and nutritive skin microcirculation in obese adolescents. METHODS: Obese female adolescents (n = 17), 15.4 +/- 2.0 years and BMI of 35.6 +/- 3.3 kg/m2, were randomized 1:1 in two groups with the diet supplemented either with Brazil nuts [BNG, n = 08, 15-25 g/day (equivalent to 3 to 5 units/day)] or placebo [PG (lactose), n = 09, one capsule/day] and followed for 16 weeks. Anthropometry, metabolic-lipid profiles, oxidative stress and morphological (capillary diameters) and functional [functional capillary density, red blood cell velocity (RBCV) at baseline and peak (RBCVmax) and time (TRBCVmax) to reach it during post occlusive reactive hyperemia, after 1 min arterial occlusion] microvascular variables were assessed by nailfold videocapillaroscopy at baseline (T0) and after intervention (T1). RESULTS: T0 characteristics were similar between groups. At T1, BNG (intra-group variation) had increased selenium levels (p = 0.02), RBCV (p = 0.03) and RBCVmax (p = 0.03) and reduced total (TC) (p = 0.02) and LDL cholesterol (p = 0.02). Compared to PG, Brazil nuts intake reduced TC (p = 0.003), triglycerides (p = 0.05) and LDL-ox (p = 0.02) and increased RBCV (p = 0.03). CONCLUSION: Brazil nuts intake improved the lipid profile and microvascular function in obese adolescents, possibly due to its high level of unsaturated fatty acids and bioactive substances. TRIAL REGISTRATION: Clinical Trials.gov NCT00937599. PMID- 21619691 TI - Autosomal dominant cerebellar ataxia type I: a review of the phenotypic and genotypic characteristics. AB - Type I autosomal dominant cerebellar ataxia (ADCA) is a type of spinocerebellar ataxia (SCA) characterized by ataxia with other neurological signs, including oculomotor disturbances, cognitive deficits, pyramidal and extrapyramidal dysfunction, bulbar, spinal and peripheral nervous system involvement. The global prevalence of this disease is not known. The most common type I ADCA is SCA3 followed by SCA2, SCA1, and SCA8, in descending order. Founder effects no doubt contribute to the variable prevalence between populations. Onset is usually in adulthood but cases of presentation in childhood have been reported. Clinical features vary depending on the SCA subtype but by definition include ataxia associated with other neurological manifestations. The clinical spectrum ranges from pure cerebellar signs to constellations including spinal cord and peripheral nerve disease, cognitive impairment, cerebellar or supranuclear ophthalmologic signs, psychiatric problems, and seizures. Cerebellar ataxia can affect virtually any body part causing movement abnormalities. Gait, truncal, and limb ataxia are often the most obvious cerebellar findings though nystagmus, saccadic abnormalities, and dysarthria are usually associated. To date, 21 subtypes have been identified: SCA1-SCA4, SCA8, SCA10, SCA12-SCA14, SCA15/16, SCA17-SCA23, SCA25, SCA27, SCA28 and dentatorubral pallidoluysian atrophy (DRPLA). Type I ADCA can be further divided based on the proposed pathogenetic mechanism into 3 subclasses: subclass 1 includes type I ADCA caused by CAG repeat expansions such as SCA1-SCA3, SCA17, and DRPLA, subclass 2 includes trinucleotide repeat expansions that fall outside of the protein-coding regions of the disease gene including SCA8, SCA10 and SCA12. Subclass 3 contains disorders caused by specific gene deletions, missense mutation, and nonsense mutation and includes SCA13, SCA14, SCA15/16, SCA27 and SCA28. Diagnosis is based on clinical history, physical examination, genetic molecular testing, and exclusion of other diseases. Differential diagnosis is broad and includes secondary ataxias caused by drug or toxic effects, nutritional deficiencies, endocrinopathies, infections and post infection states, structural abnormalities, paraneoplastic conditions and certain neurodegenerative disorders. Given the autosomal dominant pattern of inheritance, genetic counseling is essential and best performed in specialized genetic clinics. There are currently no known effective treatments to modify disease progression. Care is therefore supportive. Occupational and physical therapy for gait dysfunction and speech therapy for dysarthria is essential. Prognosis is variable depending on the type of ADCA and even among kindreds. PMID- 21619693 TI - Induction of Th1Immune responses following laser ablation in a murine model of colorectal liver metastases. AB - BACKGROUND: Preliminary experimental studies have suggested that the in situ destruction of tumor tissue by local laser ablation (LA) may also stimulate host immunity against cancer. We investigated local and systemic induction of immune responses after laser ablation in the setting of residual tumor. METHODS: A murine colorectal cancer (CRC) liver metastasis model was used. Selected tumors of liver CRC bearing mice and livers of mice without tumor induction were treated with LA. Liver and tumor tissues from the ablation sites and from distant sites were collected at various time points following LA and changes in CD3+ T cells and Kupffer cells (F4/80 marker) infiltration and the expression of interferon gamma (IFNgamma) were investigated by immunohistochemistry and ELISpot. Base line levels of CD3+ T cells and Kupffer cells were established in untreated mice. RESULTS: The presence of tumor induced significant accumulation of CD3+ T cells and Kupffer cells at the tumor-host interface, within the tumor vascular lakes and increased their baseline concentration within the liver parenchyma. LA of the liver induced accumulation of CD3+ T-cells and Kupffer cells at the site of injury and systemic induction of immune responses as discerned by the presence of IFNgamma secreting splenocytes. LA of liver tumors induced significant increase of CD3+ T-cells at site of injury, within normal liver parenchyma, and the tumor host interface of both ablated and distant tumors. In contrast Kupffer cells only accumulated in ablated tumors and the liver parenchyma but not in distant tumors. IFNgamma expression increased significantly in ablated tumors and showed an increasing trend in distant tumors. CONCLUSION: Laser ablation in addition to local tumor destruction induces local and systemic Th1 type immune responses which may play a significant role in inhibiting tumor recurrence from residual micrometastases or circulating tumor cells. PMID- 21619694 TI - MDM2 SNP309, gene-gene interaction, and tumor susceptibility: an updated meta analysis. AB - BACKGROUND: The tumor suppressor gene p53 is involved in multiple cellular pathways including apoptosis, transcriptional control, and cell cycle regulation. In the last decade it has been demonstrated that the single nucleotide polymorphism (SNP) at codon 72 of the p53 gene is associated with the risk for development of various neoplasms. MDM2 SNP309 is a single nucleotide T to G polymorphism located in the MDM2 gene promoter. From the time that this well characterized functional polymorphism was identified, a variety of case-control studies have been published that investigate the possible association between MDM2 SNP309 and cancer risk. However, the results of the published studies, as well as the subsequent meta-analyses, remain contradictory. METHODS: To investigate whether currently published epidemiological studies can clarify the potential interaction between MDM2 SNP309 and the functional genetic variant in p53 codon72 (Arg72Pro) and p53 mutation status, we performed a meta-analysis of the risk estimate on 27,813 cases with various tumor types and 30,295 controls. RESULTS: The data we reviewed indicated that variant homozygote 309GG and heterozygote 309TG were associated with a significant increased risk of all tumor types (homozygote comparison: odds ratio (OR) = 1.25, 95% confidence interval (CI) = 1.13-1.37; heterozygote comparison: OR = 1.10, 95% CI = 1.03-1.17). We also found that the combination of GG and Pro/Pro, TG and Pro/Pro, GG and Arg/Arg significantly increased the risk of cancer (OR = 3.38, 95% CI = 1.77-6.47; OR = 1.88, 95% CI = 1.26-2.81; OR = 1.96, 95% CI = 1.01-3.78, respectively). In a stratified analysis by tumor location, we also found a significant increased risk in brain, liver, stomach and uterus cancer (OR = 1.47, 95% CI = 1.06-2.03; OR = 2.24, 95%CI = 1.57-3.18; OR = 1.54, 95%CI = 1.04-2.29; OR = 1.34, 95%CI = 1.07 1.29, respectively). However, no association was seen between MDM2 SNP309 and tumor susceptibility in the stratified analysis by p53 mutation status (GG vs TT: OR = 1.17, 95% CI = 0.75-1.82 and TG vs TT: OR = 1.09, 95% CI = 0.89-1.34 for positive p53 mutation status; GG vs TT: OR = 0.95, 95% CI = 0.72-1.25 and TG vs TT: OR = 1.06, 95% CI = 0.85-1.30 for negative p53 mutation status). CONCLUSIONS: The analyses indicate that MDM2 SNP309 serves as a tumor susceptibility marker, and that there is an association between MDM2 SNP309 and p53 Arg72Pro regarding tumor susceptibility. Further studies that take into consideration environmental stresses and functional genetic variants in the p53-MDM2-related genes are warranted. PMID- 21619695 TI - Social networks, work and network-based resources for the management of long-term conditions: a framework and study protocol for developing self-care support. AB - BACKGROUND: Increasing the effective targeting and promotion of self-care support for long-term conditions requires more of a focus on patient contexts and networks. The aim of this paper is to describe how within a programme of research and implementation, social networks are viewed as being centrally involved in the mobilisation and deployment of resources in the management of a chronic condition. This forms the basis of a novel approach to understanding, designing, and implementing new forms of self-management support. METHODS: Drawing on evidence syntheses about social networks and capital and the role of information in self-management, we build on four conceptual approaches to inform the design of our research on the implementation of self-care support for people with long term conditions. Our approach takes into consideration the form and content of social networks, notions of chronic illness work, normalisation process theory (NPT), and the whole systems informing self-management engagement (WISE) approach to self-care support. DISCUSSION: The translation and implementation of a self care agenda in contemporary health and social context needs to acknowledge and incorporate the resources and networks operating in patients' domestic and social environments and everyday lives. The latter compliments the focus on healthcare settings for developing and delivering self-care support by viewing communities and networks, as well as people suffering from long-term conditions, as a key means of support for managing long-term conditions. By focusing on patient work and social-network provision, our aim is to open up a second frontier in implementation research, to translate knowledge into better chronic illness management, and to shift the emphasis towards support that takes place outside formal health services. PMID- 21619696 TI - Conotoxin protein classification using free scores of words and support vector machines. AB - BACKGROUND: Conotoxin has been proven to be effective in drug design and could be used to treat various disorders such as schizophrenia, neuromuscular disorders and chronic pain. With the rapidly growing interest in conotoxin, accurate conotoxin superfamily classification tools are desirable to systematize the increasing number of newly discovered sequences and structures. However, despite the significance and extensive experimental investigations on conotoxin, those tools have not been intensively explored. RESULTS: In this paper, we propose to consider suboptimal alignments of words with restricted length. We developed a scoring system based on local alignment partition functions, called free score. The scoring system plays the key role in the feature extraction step of support vector machine classification. In the classification of conotoxin proteins, our method, SVM-Freescore, features an improved sensitivity and specificity by approximately 5.864% and 3.76%, respectively, over previously reported methods. For the generalization purpose, SVM-Freescore was also applied to classify superfamilies from curated and high quality database such as ConoServer. The average computed sensitivity and specificity for the superfamily classification were found to be 0.9742 and 0.9917, respectively. CONCLUSIONS: The SVM-Freescore method is shown to be a useful sequence-based analysis tool for functional and structural characterization of conotoxin proteins. The datasets and the software are available at http://faculty.uaeu.ac.ae/nzaki/SVM-Freescore.htm. PMID- 21619697 TI - Political activity for physical activity: health advocacy for active transport. AB - Effective health advocacy is a priority for efforts to increase population participation in physical activity. Local councils are an important audience for this advocacy. The aim of the current study was to describe features of advocacy for active transport via submissions to city council annual plans in New Zealand, and the impact of an information sheet to encourage the health sector to be involved in this process. Written submissions to city council's annual consultation process were requested for 16 city councils over the period of three years (2007/08, 2008/09, and 2009/10). Submissions were reviewed and categories of responses were created. An advocacy information sheet encouraging health sector participation and summarising some of the evidence-base related to physical activity, active transport and health was released just prior to the 2009/10 submission time. Over the period of the study, city councils received 47,392 submissions, 17% of which were related to active transport. Most submissions came from city residents, with a small proportion (2%) from the health sector. The largest category of submissions was in support of pedestrian and cycling infrastructure, design and maintenance of facilities and additional features to support use of these transport modes. Health arguments featured prominently in justifications for active transport initiatives, including concerns about injury risk, obesity, physical inactivity, personal safety and facilities for people with disabilities. There was evidence that the information sheet was utilised by some health sector submitters (12.5%), providing tentative support for initiatives of this nature. In conclusion, the study provides novel information about the current nature of health advocacy for active transport and informs future advocacy efforts about areas for emphasis, such as health benefits of active transport, and potential alliances with other sectors such as environmental sustainability, transport and urban planning and local communities. PMID- 21619698 TI - A novel family VII esterase with industrial potential from compost metagenomic library. AB - BACKGROUND: Among the vast microbial genomic resources now available, most microbes are unculturable in the laboratory. A culture-independent metagenomic approach is a novel technique that circumvents this culture limitation. For the screening of novel lipolytic enzymes, a metagenomic library was constructed from compost, and the clone of estCS2 was selected for lipolytic properties on a tributyrin-containing medium. RESULTS: The estCS2 sequence encodes a protein of 570 amino acid residues, with a predicted molecular mass of 63 kDa, and based on amino acid identity it most closely matches (45%) the carboxylesterase from Haliangium ochraceum DSM 14365. EstCS2 belong to family VII, according to the lipolytic enzyme classification proposed by Arpigny and Jaeger, and it retains the catalytic triad Ser245-Glu363-His466 that is typical of an alpha/beta hydrolase. The Ser245 residue in the catalytic triad of EstCS2 is located in the consensus active site motif GXSXG. The EstCS2 exhibits strong activity toward p nitrophenyl caproate (C6), and it is stable up to 60 degrees C with an optimal enzymatic activity at 55 degrees C. The maximal activity is observed at pH 9, and it remains active between pH 6-10. EstCS2 shows remarkable stability in up to 50% (v/v) dimethyl sulfoxide (DMSO) or dimethylformamide (DMF). The enzyme has the ability to cleave sterically hindered esters of tertiary alcohol, as well as to degrade polyurethanes, which are widely used in various industries. CONCLUSIONS: The high stability of EstCS2 in organic solvents and its activity towards esters of ketoprofen and tertiary alcohols, and in polyurethane suggests that it has potential uses for many applications in biotransformation and bioremediation. PMID- 21619699 TI - Dermatan sulfate in tunicate phylogeny: order-specific sulfation pattern and the effect of [->4IdoA(2-sulfate)beta-1->3GalNAc(4-sulfate)beta-1->] motifs in dermatan sulfate on heparin cofactor II activity. AB - BACKGROUND: Previously, we have reported the presence of highly sulfated dermatans in solitary ascidians from the orders Phlebobranchia (Phallusia nigra) and Stolidobranchia (Halocynthia pyriformis and Styela plicata). Despite the identical disaccharide backbone, consisting of [->4IdoA(2S)beta-1->3GalNAcbeta-1 >], those polymers differ in the position of sulfation on the N-Acetyl galactosamine, which can occur at carbon 4 or 6. We have shown that position rather than degree of sulfation is important for heparin cofactor II activity. As a consequence, 2,4- and 2,6-sulfated dermatans have high and low heparin cofactor II activities, respectively. In the present study we extended the disaccharide analysis of ascidian dermatan sulfates to additional species of the orders Stolidobranchia (Herdmania pallida, Halocynthia roretzi) and Phlebobranchia (Ciona intestinalis), aiming to investigate how sulfation evolved within Tunicata. In addition, we analysed how heparin cofactor II activity responds to dermatan sulfates containing different proportions of 2,6- or 2,4-disulfated units. RESULTS: Disaccharide analyses indicated a high content of disulfated disaccharide units in the dermatan sulfates from both orders. However, the degree of sulfation decreased from Stolidobranchia to Phlebobranchia. While 76% of the disaccharide units in dermatan sulfates from stolidobranch ascidians are disulfated, 53% of disulfated disaccharides are found in dermatan sulfates from phlebobranch ascidians. Besides this notable difference in the sulfation degree, dermatan sulfates from phlebobranch ascidians contain mainly 2,6-sulfated disaccharides whereas dermatan sulfate from the stolidobranch ascidians contain mostly 2,4-sulfated disaccharides, suggesting that the biosynthesis of dermatan sulfates might be differently regulated during tunicates evolution. Changes in the position of sulfation on N-acetylgalactosamine in the disaccharide [->4IdoA(2 Sulfate)beta-1->3GalNAcbeta-1->] modulate heparin cofactor II activity of dermatan sulfate polymers. Thus, high and low heparin cofactor II stimulating activity is observed in 2,4-sulfated dermatan sulfates and 2,6-sulfated dermatan sulfates, respectively, confirming the clear correlation between the anticoagulant activities of dermatan sulfates and the presence of 2,4-sulfated units. CONCLUSIONS: Our results indicate that in ascidian dermatan sulfates the position of sulfation on the GalNAc in the disaccharide [->4IdoA(2S)beta-1 >3GalNAcbeta-1->] is directly related to the taxon and that the 6-O sulfation is a novelty apparently restricted to the Phlebobranchia. We also show that the increased content of [->4IdoA(2S)beta-1->3GalNAc(4S)beta-1->] disaccharide units in dermatan sulfates from Stolidobranchia accounts for the increased heparin cofactor II stimulating activity. PMID- 21619700 TI - In vitro interactions of Candida parapsilosis wild type and lipase deficient mutants with human monocyte derived dendritic cells. AB - BACKGROUND: Candida parapsilosis typically is a commensal of human skin. However, when host immune defense is compromised or the normal microflora balance is disrupted, C. parapsilosis transforms itself into an opportunistic pathogen. Candida-derived lipase has been identified as potential virulence factor. Even though cellular components of the innate immune response, such as dendritic cells, represent the first line of defense against invading pathogens, little is known about the interaction of these cells with invading C. parapsilosis. Thus, the aim of our study was to assess the function of dendritic cells in fighting C. parapsilosis and to determine the role that C. parapsilosis-derived lipase plays in the interaction with dendritic cells. RESULTS: Monocyte-derived immature and mature dendritic cells (iDCs and mDCs, respectively) co-cultured with live wild type or lipase deficient C. parapsilosis strains were studied to determine the phagocytic capacity and killing efficiency of host cells. We determined that both iDCs and mDCs efficiently phagocytosed and killed C. parapsilosis, furthermore our results show that the phagocytic and fungicidal activities of both iDCs and mDCs are more potent for lipase deficient compared to wild type yeast cells. In addition, the lipase deficient C. parapsilosis cells induce higher gene expression and protein secretion of proinflammatory cytokines and chemokines in both DC types relative to the effect of co-culture with wild type yeast cells. CONCLUSIONS: Our results show that DCs are activated by exposure to C. parapsilosis, as shown by increased phagocytosis, killing and proinflammatory protein secretion. Moreover, these data strongly suggest that C. parapsilosis derived lipase has a protective role during yeast:DC interactions, since lipase production in wt yeast cells decreased the phagocytic capacity and killing efficiency of host cells and downregulated the expression of host effector molecules. PMID- 21619701 TI - Health-related quality of life of young adults with Turner syndrome following a long-term randomized controlled trial of recombinant human growth hormone. AB - BACKGROUND: There are limited long-term randomized controlled trials of growth hormone (GH) supplementation to adult height and few published reports of the health-related quality of life (HRQOL) following treatment. The present follow-up study of young adults from a long-term controlled trial of GH treatment in patients with Turner syndrome (TS) yielded data to examine whether GH supplementation resulted in a higher HRQOL (either due to taller stature or from the knowledge that active treatment and not placebo had been received) or alternatively a lower HRQOL (due to medicalization from years of injections). METHODS: The original trial randomized 154 Canadian girls with TS aged 7-13 years from 13 centres to receive either long-term GH injections at the pharmacologic dose of 0.3 mg/kg/week or to receive no injections; estrogen prescription for induction of puberty was standardized. Patients were eligible for the follow-up study if they were at least 16 years old at the time of follow-up. The instrument used to study HRQOL was the SF-36, summarized into physical and mental component scales (PCS and MCS); higher scores indicate better HRQOL. RESULTS: Thirty-four of the 48 eligible participants (71%) consented to participate; data were missing for one patient. Both groups (GH and no treatment) had normal HRQOL at this post treatment assessment. The GH group had a (mean +/- SD) PCS score of 56 +/- 5; the untreated group 58 +/- 4; mean score for 16-24 year old females in the general population 53.5 +/- 6.9. The GH group had a mean MCS score of 52 +/- 6; the untreated group 49 +/- 13; mean score for 16-24 year old females in the general population 49.6 +/- 9.8. Secondary analyses showed no relationship between HRQOL and height. CONCLUSIONS: We found no benefit or adverse effect on HRQOL either from receiving or not receiving growth hormone injections in a long-term randomized controlled trial, confirming larger observational studies. We suggest that it remains ethically acceptable as well as necessary to maintain a long-term untreated control group to estimate the effects of pharmacological agents to manipulate adult height. Young adult women with TS have normal HRQOL suggesting that they adjust well to their challenges in life. TRIAL REGISTRATION: ClinicalTrials.gov Identifier NCT00191113. PMID- 21619702 TI - Growth of a cohort of very low birth weight infants in Johannesburg, South Africa. AB - BACKGROUND: Little is known about the growth of VLBW infants in South Africa. The aim of this study was to assess the growth of a cohort of VLBW infants in Johannesburg. METHODS: A secondary analysis of a prospective cohort was conducted on 139 VLBW infants (birth weight <= 1500 g) admitted to Charlotte Maxeke Johannesburg Academic Hospital. Growth measurements were obtained from patient files and compared with the World Health Organization Child Growth Standards (WHO CGS) and with a previous cohort of South African VLBW infants. The sample size per analysis ranged from 11 to 81 infants. RESULTS: Comparison with the WHO-CGS showed initial poor growth followed by gradual catch up growth with mean Z scores of 0.0 at 20 months postmenstrual age for weight, -0.8 at 20 months postmenstrual age for length and 0.0 at 3 months postmenstrual age for head circumference. Growth was comparable with that of a previous cohort of South African VLBW infants in all parameters. CONCLUSIONS: Initial poor growth in the study sample was followed by gradual catch up growth but with persistent deficits in length for age at 20 months postmenstrual age relative to healthy term infants. PMID- 21619703 TI - OntoCAT--simple ontology search and integration in Java, R and REST/JavaScript. AB - BACKGROUND: Ontologies have become an essential asset in the bioinformatics toolbox and a number of ontology access resources are now available, for example, the EBI Ontology Lookup Service (OLS) and the NCBO BioPortal. However, these resources differ substantially in mode, ease of access, and ontology content. This makes it relatively difficult to access each ontology source separately, map their contents to research data, and much of this effort is being replicated across different research groups. RESULTS: OntoCAT provides a seamless programming interface to query heterogeneous ontology resources including OLS and BioPortal, as well as user-specified local OWL and OBO files. Each resource is wrapped behind easy to learn Java, Bioconductor/R and REST web service commands enabling reuse and integration of ontology software efforts despite variation in technologies. It is also available as a stand-alone MOLGENIS database and a Google App Engine application. CONCLUSIONS: OntoCAT provides a robust, configurable solution for accessing ontology terms specified locally and from remote services, is available as a stand-alone tool and has been tested thoroughly in the ArrayExpress, MOLGENIS, EFO and Gen2Phen phenotype use cases. AVAILABILITY: http://www.ontocat.org. PMID- 21619704 TI - A comprehensive candidate gene approach identifies genetic variation associated with osteosarcoma. AB - BACKGROUND: Osteosarcoma (OS) is a bone malignancy which occurs primarily in adolescents. Since it occurs during a period of rapid growth, genes important in bone formation and growth are plausible modifiers of risk. Genes involved in DNA repair and ribosomal function may contribute to OS pathogenesis, because they maintain the integrity of critical cellular processes. We evaluated these hypotheses in an OS association study of genes from growth/hormone, bone formation, DNA repair, and ribosomal pathways. METHODS: We evaluated 4836 tag SNPs across 255 candidate genes in 96 OS cases and 1426 controls. Logistic regression models were used to estimate the odds ratios (OR) and 95% confidence intervals (CI). RESULTS: Twelve SNPs in growth or DNA repair genes were significantly associated with OS after Bonferroni correction. Four SNPs in the DNA repair gene FANCM (ORs 1.9-2.0, P = 0.003-0.004) and 2 SNPs downstream of the growth hormone gene GH1 (OR 1.6, P = 0.002; OR 0.5, P = 0.0009) were significantly associated with OS. One SNP in the region of each of the following genes was significant: MDM2, MPG, FGF2, FGFR3, GNRH2, and IGF1. CONCLUSIONS: Our results suggest that several SNPs in biologically plausible pathways are associated with OS. Larger studies are required to confirm our findings. PMID- 21619705 TI - Large-scale analysis of expression signatures reveals hidden links among diverse cellular processes. AB - BACKGROUND: Cells must respond to various perturbations using their limited available gene repertoires. In order to study how cells coordinate various responses, we conducted a comprehensive comparison of 1,186 gene expression signatures (gene lists) associated with various genetic and chemical perturbations. RESULTS: We identified 7,419 statistically significant overlaps between various published gene lists. Most (80%) of the overlaps can be represented by a highly connected network, a "molecular signature map," that highlights the correlation of various expression signatures. By dissecting this network, we identified sub-networks that define clusters of gene sets related to common biological processes (cell cycle, immune response, etc). Examination of these sub-networks has confirmed relationships among various pathways and also generated new hypotheses. For example, our result suggests that glutamine deficiency might suppress cellular growth by inhibiting the MYC pathway. Interestingly, we also observed 1,369 significant overlaps between a set of genes upregulated by factor X and a set of genes downregulated by factor Y, suggesting a repressive interaction between X and Y factors. CONCLUSIONS: Our results suggest that molecular-level responses to diverse chemical and genetic perturbations are heavily interconnected in a modular fashion. Also, shared molecular pathways can be identified by comparing newly defined gene expression signatures with databases of previously published gene expression signatures. PMID- 21619706 TI - Drug-related admissions and hospital-acquired adverse drug events in Germany: a longitudinal analysis from 2003 to 2007 of ICD-10-coded routine data. AB - BACKGROUND: Adverse reactions and medication errors are complications of drug use. Spontaneous reporting systems and pharmacoepidemiological studies incompletely detect the occurrence of these events in daily hospital care. In this study, the frequency and type of drug-related admissions and hospital acquired adverse drug events (ADE) in Germany were assessed using routinely collected hospital data. METHODS: The study was based on aggregated hospital routine data covering the period 2003 to 2007 and annually recorded as part of the further development of the German Diagnosis-Related Groups. The 505 ICD-10 codes indicating an ADE were categorized in seven groups according to their certainty. Primary diagnoses were considered as a proxy for drug-related admissions, and secondary diagnoses as a proxy for hospital-acquired ADE. RESULTS: Among all hospital admissions, 5% were found to be at least possibly drug-induced and 0.7% very likely drug-induced. There was a significant increase in the overall rate of drug-related admissions over time (p < 0.038). Enterocolitis due to Clostridium difficile infection was the most frequent cause of a drug-related admission. About 4.5% of in-patients had experienced a hospital acquired ADE. In addition, over the course of the study period, the overall frequency of hospital-acquired ADEs significantly increased (p < 0.001). CONCLUSIONS: In Germany, more than 5% of hospital episodes are either caused or complicated by an ADE. Between 2003 and 2007, there was a statistically significant increase in the overall rate and in some of the subcategories defined by the list of ICD-10-codes suspected to be indicative of an ADE. Before the use of routine data in pharmacovigilance and patient safety can be fully exploited, a further tailoring of both the ICD and the available variable set is needed. PMID- 21619707 TI - Evaluation of internet access and utilization by medical students in Lahore, Pakistan. AB - BACKGROUND: The internet is increasingly being used worldwide in imparting medical education and improving its delivery. It has become an important tool for healthcare professionals training but the data on its use by medical students in developing countries is lacking with no study on the subject from Pakistan. This study was, therefore, carried out with an aim to evaluate the pattern of internet access and utilization by medical students in Pakistan. METHODS: A structured pre tested questionnaire was administered to a group of 750 medical students in clinical years studying at various public and private medical colleges in Lahore. The questions were related to patterns of internet access, purpose of use and self reported confidence in performing various internet related tasks, use of health related websites to supplement learning and the problems faced by students in using internet at the institution. RESULTS: A total of 532 medical students (70.9%) returned the questionnaire. The mean age of study participants was 21.04 years (SD 1.96 years). Majority of the respondents (84.0%) reported experience with internet use. About half of the students (42.1%) were using internet occasionally with 23.1%, 20.9% and 13.9% doing so frequently, regularly and rarely respectively. About two third of the students (61.0%) stated that they use internet for both academic and professional activities. Most of the participants preferred to use internet at home (70.5%). Self reported ability to search for required article from PubMed and PakMedinet was reported by only 34.0% of the entire sample. Students were moderately confident in performing various internet related tasks including downloading medical books from internet, searching internet for classification of diseases and downloading full text article. Health related websites were being accessed by 55.1% students to supplement their learning process. Lack of time, inadequate number of available computers and lack of support from staff were cited as the most common problems faced by students while accessing internet in the institution premises. There were significant differences among male and female students with respect to the place of internet use (p = 0.001) and the ability to search online databases for required articles (p = 0.014). CONCLUSIONS: Majority of the medical students in this study had access to internet and were using it for both academic and personal reasons. Nevertheless, it was seen that there is under utilization of the potential of internet resources to augment learning. Increase in awareness, availability of requisite facilities and training in computing skills are required to enable better utilization of digital resources of digital resources by medical students. PMID- 21619708 TI - Nutritional management of Eosinophilic Gastroenteropathies: Case series from the community. AB - Eosinophilic gastroenteropathies, such as eosinophilic esophagitis and eosinophilic colitis, have classically been treated with swallowed inhaled corticosteroids or oral corticosteroids. More recent studies have found elimination and elemental diets to be effective treatment alternatives to steroids. In this case series we describe the treatment of three children using nutritional management in a community setting. Elimination diets and elemental diets based on patch testing and skin prick tests reduced the eosinophil counts to normal levels in all three children. Food items which tested positive were then reintroduced while symptoms and eosinophil counts were monitored. Nutritional management of eosinophilic esophagitis and eosinophilic colitis was found to be effective in reducing symptoms. However, obstacles facing patients who choose this type of therapy include limitations due to the cost of repeated endoscopies, palatability of elimination/elemental diets and the availability of subspecialists trained in management (e.g. Allergy, Gastroenterology, and Pathology). It may be a worthwhile endeavour to overcome these obstacles as nutritional management minimizes the potential long-term effects of chronic steroid therapy. PMID- 21619709 TI - Prognostic significance of IL-6 and IL-8 ascites levels in ovarian cancer patients. AB - BACKGROUND: The acellular fraction of epithelial ovarian cancer (EOC) ascites promotes de novo resistance of tumor cells and thus supports the idea that tumor cells may survive in the surrounding protective microenvironment contributing to disease recurrence. Levels of the pro-inflammatory cytokines IL-6 and IL-8 are elevated in EOC ascites suggesting that they could play a role in tumor progression. METHODS: We measured IL-6 and IL-8 levels in the ascites of 39 patients with newly diagnosed EOC. Commercially available enzyme-linked immunosorbent assay (ELISA) was used to determine IL-6 and IL-8 ascites levels. Ascites cytokine levels were correlated with clinicopathological parameters and progression-free survival. RESULTS: Mean ascites levels for IL-6 and IL-8 were 6419 pg/ml (SEM: 1409 pg/ml) and 1408 pg/ml (SEM: 437 pg/ml) respectively. The levels of IL-6 and IL-8 in ascites were significantly lower in patients that have received prior chemotherapy before the surgery (Mann-Whitney U test, P = 0.037 for IL-6 and P = 0.008 for IL-8). Univariate analysis revealed that high IL-6 ascites levels (P = 0.021), serum CA125 levels (P = 0.04) and stage IV (P = 0.009) were significantly correlated with shorter progression-free survival. Including these variables in a multivariate analysis revealed that elevated IL-6 levels (P = 0.033) was an independent predictor of shorter progression-free survival. CONCLUSION: Elevated IL-6, but not IL-8, ascites level is an independent predictor of shorter progression-free survival. PMID- 21619710 TI - Lower limb biomechanics during running in individuals with achilles tendinopathy: a systematic review. AB - BACKGROUND: Abnormal lower limb biomechanics is speculated to be a risk factor for Achilles tendinopathy. This study systematically reviewed the existing literature to identify, critique and summarise lower limb biomechanical factors associated with Achilles tendinopathy. METHODS: We searched electronic bibliographic databases (Medline, EMBASE, Current contents, CINAHL and SPORTDiscus) in November 2010. All prospective cohort and case-control studies that evaluated biomechanical factors (temporospatial parameters, lower limb kinematics, dynamic plantar pressures, kinetics [ground reaction forces and joint moments] and muscle activity) associated with mid-portion Achilles tendinopathy were included. Quality of included studies was evaluated using the Quality Index. The magnitude of differences (effect sizes) between cases and controls was calculated using Cohen's d (with 95% CIs). RESULTS: Nine studies were identified; two were prospective and the remaining seven case-control study designs. The quality of 9 identified studies was varied, with Quality Index scores ranging from 4 to 15 out of 17. All studies analysed running biomechanics. Cases displayed increased eversion range of motion of the rearfoot (d = 0.92 and 0.67 in two studies), reduced maximum lower leg abduction (d = -1.16), reduced ankle joint dorsiflexion velocity (d = -0.62) and reduced knee flexion during gait (d = -0.90). Cases also demonstrated a number of differences in dynamic plantar pressures (primarily the distribution of the centre of force), ground reaction forces (large effects for timing variables) and also showed reduced peak tibial external rotation moment (d = -1.29). Cases also displayed differences in the timing and amplitude of a number of lower limb muscles but many differences were equivocal. CONCLUSIONS: There are differences in lower limb biomechanics between those with and without Achilles tendinopathy that may have implications for the prevention and management of the condition. However, the findings need to be interpreted with caution due to the limited quality of a number of the included studies. Future well-designed prospective studies are required to confirm these findings. PMID- 21619711 TI - Proteomic analysis identifies interleukin 11 regulated plasma membrane proteins in human endometrial epithelial cells in vitro. AB - BACKGROUND: During the peri-implantation period, the embryo adheres to an adequately prepared or receptive endometrial surface epithelium. Abnormal embryo adhesion to the endometrium results in embryo implantation failure and infertility. Endometrial epithelial cell plasma membrane proteins critical in regulating adhesion may potentially be infertility biomarkers or targets for treating infertility. Interleukin (IL) 11 regulates human endometrial epithelial cells (hEEC) adhesion. Its production is abnormal in women with infertility. The objective of the study was to identify IL11 regulated plasma membrane proteins in hEEC in vitro using a proteomic approach. METHODS: Using a 2D-differential in-gel electrophoresis (DIGE) electrophoresis combined with LCMS/MS mass spectrometry approach, we identified 20 unique plasma membrane proteins differentially regulated by IL11 in ECC-1 cells, a hEEC derived cell line. Two IL11 regulated proteins with known roles in cell adhesion, annexin A2 (ANXA2) and flotillin-1 (FLOT1), were validated by Western blot and immunocytochemistry in hEEC lines (ECC-1 and an additional cell line, Ishikawa) and primary hEEC. Flotilin-1 was further validated by immunohistochemistry in human endometrium throughout the menstrual cycle (n = 6-8/cycle). RESULTS: 2D-DIGE analysis identified 4 spots that were significantly different between control and IL11 treated group. Of these 4 spots, there were 20 proteins that were identified with LCMS/MS. Two proteins; ANXA2 and FLOT1 were chosen for further analyses and have found to be significantly up-regulated following IL11 treatment. Western blot analysis showed a 2-fold and a 2.5-fold increase of ANXA2 in hEEC membrane fraction of ECC-1 and Ishikawa cells respectively. Similarly, a 1.8-fold and a 2.3/2.4-fold increase was also observed for FLOT1 in hEEC membrane fraction of ECC-1 and Ishikawa cells respectively. In vitro, IL11 induced stronger ANXA2 expression on cell surface of primary hEEC and ECC-1 whilst, the lipid-raft protein FLOT1 demonstrated punctate staining in the apical surface of ECC-1 plasma membranes and was upregulated in the epithelium in the receptive phase of the menstrual cycle (p lower or equal 0.05). CONCLUSIONS: This is the first study to use a proteomics approach to identify hEEC plasma membrane proteins that may be useful as infertility markers or pharmacological targets for fertility regulation. PMID- 21619712 TI - Identification of CD8+ cytotoxic T lymphocyte epitopes from porcine reproductive and respiratory syndrome virus matrix protein in BALB/c mice. AB - Twenty-seven nanopeptides derived from the matrix (M) protein of porcine reproductive and respiratory syndrome virus (PRRSV) were screened for their ability to elicit a recall interferon-gamma (IFN-gamma) response from the splenocytes of BALB/c mice following DNA vaccination and a booster vaccination with recombinant vaccinia virus rWR-PRRSV-M. We identified two peptides (amino acid residues K93FITSRCRL and F57GYMTFVHF) as CD8+ cytotoxic T lymphocyte (CTL) epitopes. These peptides elicited significant numbers of IFN-gamma secreting cells, compared with other M nonapeptides and one irrelevant nonapeptide. Bioinformatics analysis showed that the former is an H-2Kd-restricted CTL epitope, and the latter is an H-2Dd-restricted CTL epitope. Multiple amino acid sequence alignment among different PRRSV M sequences submitted to GenBank indicated that these two CTL epitopes are strongly conserved, and they should therefore be considered for further research on the mechanisms of cellular immune responses to PRRSV. PMID- 21619713 TI - Intragenomic diversity of Rhizobium leguminosarum bv. trifolii clover nodule isolates. AB - BACKGROUND: Soil bacteria from the genus Rhizobium are characterized by a complex genomic architecture comprising chromosome and large plasmids. Genes responsible for symbiotic interactions with legumes are usually located on one of the plasmids, named the symbiotic plasmid (pSym). The plasmids have a great impact not only on the metabolic potential of rhizobia but also underlie genome rearrangements and plasticity. RESULTS: Here, we analyzed the distribution and sequence variability of markers located on chromosomes and extrachromosomal replicons of Rhizobium leguminosarum bv. trifolii strains originating from nodules of clover grown in the same site in cultivated soil. First, on the basis of sequence similarity of repA and repC replication genes to the respective counterparts of chromids reported in R. leguminosarum bv. viciae 3841 and R. etli CFN42, chromid-like replicons were distinguished from the pool of plasmids of the nodule isolates studied. Next, variability of the gene content was analyzed in the different genome compartments, i.e., the chromosome, chromid-like and 'other plasmids'. The stable and unstable chromosomal and plasmid genes were detected on the basis of hybridization data. Displacement of a few unstable genes between the chromosome, chromid-like and 'other plasmids', as well as loss of some markers was observed in the sampled strains. Analyses of chosen gene sequences allowed estimation of the degree of their adaptation to the three genome compartments as well as to the host. CONCLUSIONS: Our results showed that differences in distribution and sequence divergence of plasmid and chromosomal genes can be detected even within a small group of clover nodule isolates recovered from clovers grown at the same site. Substantial divergence of genome organization could be detected especially taking into account the content of extrachromosomal DNA. Despite the high variability concerning the number and size of plasmids among the studied strains, conservation of the location as well as dynamic distribution of the individual genes (especially replication genes) of a particular genome compartment were demonstrated. The sequence divergence of particular genes may be affected by their location in the given genome compartment. The 'other plasmid' genes are less adapted to the host genome than the chromosome and chromid-like genes. PMID- 21619714 TI - Characterisation of a novel paralog of scavenger receptor class B member I (SCARB1) in Atlantic salmon (Salmo salar). AB - BACKGROUND: Red flesh colour is a unique trait found in some salmonid genera. Carotenoid pigments are not synthesized de novo in the fish, but are provided by dietary uptake. A better understanding of the molecular mechanisms underlying the cellular uptake and deposition of carotenoids could potentially be used to improve the low muscle deposition rate that is typically found in farmed Atlantic salmon. In addition, from an evolutionary point of view, the establishment and maintenance of this trait is still poorly understood. It has been demonstrated in several species that scavenger receptor class B, member 1 (SCARB1) is involved in intestinal absorption of carotenoids, which makes this gene a possible source of genetic variation in salmonid flesh pigmentation. RESULTS: In this study, a novel paralog of SCARB1 (SCARB1-2) was detected through screening for genetic variation in Atlantic salmon SCARB1. Full length SCARB1-2 encodes a protein with 89% identity to Atlantic salmon SCARB1, except for the C-terminal cytoplasmic tail that shows only 12% identity. The most prominent site of SCARB1 mRNA expression was in the mid gut, while a five-fold lower level was detected in Atlantic salmon skeletal muscle and liver. The SCARB1-2 mRNA was equally expressed in liver, muscle and mid gut, and at a lower level than SCARB1 mRNA. A total of seven different SCARB1-2 alleles comprising repetitive enhancer of zeste motifs (EZH2) were identified in the founding parents of a resource Atlantic salmon population. We mapped the SCARB1-2 paralog to a region on Atlantic salmon chromosome 1, containing a putative QTL for flesh colour. Addition of the SCARB1-2 marker increased the significance of this QTL, however the large confidence interval surrounding the QTL precludes confirmation of SCARB1-2 as a causative gene underlying variation in this trait. CONCLUSION: We have characterised a novel paralog of SCARB1 (SCARB1-2), have mapped it to Atlantic salmon chromosome 1 and have described its expression in various tissues. Mapping with SCARB1-2 alleles added further evidence for a QTL affecting flesh colour on this chromosome, however further studies are needed to confirm a functional role for this gene in flesh colour pigmentation. PMID- 21619715 TI - Biomarkers of oxidative stress and its association with the urinary reducing capacity in bus maintenance workers. AB - BACKGROUND: Exposure to particles (PM) induces adverse health effects (cancer, cardiovascular and pulmonary diseases). A key-role in these adverse effects seems to be played by oxidative stress, which is an excess of reactive oxygen species relative to the amount of reducing species (including antioxidants), the first line of defense against reactive oxygen species. The aim of this study was to document the oxidative stress caused by exposure to respirable particles in vivo, and to test whether exposed workers presented changes in their urinary levels for reducing species. METHODS: Bus depot workers (n = 32) exposed to particles and pollutants (respirable PM4, organic and elemental carbon, particulate metal content, polycyclic aromatic hydrocarbons, NOx, O3) were surveyed over two consecutive days. We collected urine samples before and after each shift, and quantified an oxidative stress biomarker (8-hydroxy-2'-deoxyguanosine), the reducing capacity and a biomarker of PAH exposure (1-hydroxypyrene). We used a linear mixed model to test for associations between the oxidative stress status of the workers and their particle exposure as well as with their urinary level of reducing species. RESULTS: Workers were exposed to low levels of respirable PM4 (range 25-71 MUg/m3). However, urinary levels of 8-hydroxy-2'-deoxyguanosine increased significantly within each shift and between both days for non-smokers. The between-day increase was significantly correlated (p < 0.001) with the concentrations of organic carbon, NOx, and the particulate copper content. The within-shift increase in 8OHdG was highly correlated to an increase of the urinary reducing capacity (Spearman rho = 0.59, p < 0.0001). CONCLUSIONS: These findings confirm that exposure to components associated to respirable particulate matter causes a systemic oxidative stress, as measured with the urinary 8OHdG. The strong association observed between urinary 8OHdG with the reducing capacity is suggestive of protective or other mechanisms, including circadian effects. Additional investigations should be performed to understand these observations. PMID- 21619716 TI - Disability pension by occupational class--the impact of work-related factors: the Hordaland Health Study Cohort. AB - BACKGROUND: The social gradient in disability pension is well recognized, however mechanisms accounting for the gradient are largely unknown. The aim of this study was to examine the association between occupational class and subsequent disability pension among middle-aged men and women, and to what extent work related factors accounted for the association. METHODS: A subsample (N = 7031) of the population-based Hordaland Health Study (HUSK) conducted in 1997-99, provided self-reported information on health and work-related factors, and were grouped in four strata by Erikson, Goldthorpe and Portocareros occupational class scheme. The authors obtained follow-up data on disability pension by linking the health survey to national registries of benefit (FD-trygd). They employed Cox regression analysis and adjusted for gender, health (medical conditions, mental health, self perceived health, somatic symptoms) and work-related factors (working hours, years in current occupation, physical demands, job demands, job control). RESULTS: A strong gradient in disability pension by occupational class was found. In the fully adjusted model the risk (hazard ratio) ranged from 1.41 (95% CI 0.84 to 2.33) in the routine non-manual class, 1.87 (95% CI 1.07 to 3.27) in the skilled manual class and 2.12 (95% CI 1.14 to 3.95) in the unskilled manual class, employing the administrator and professional class as reference. In the gender and health-adjusted model work-related factors mediated the impact of occupational class on subsequent disability pension with 5% in the routine non manual class, 26% in the skilled manual class and 24% in the unskilled manual class. The impact of job control and physical demands was modest, and mainly seen among skilled and unskilled manual workers. CONCLUSIONS: Workers in the skilled and unskilled manual classes had a substantial unexplained risk of disability pension. Work-related factors only had a moderate impact on the disability risk. Literature indicates an accumulation of hazards in the manual classes. This should be taken into account when interpreting the gradient in disability pension. PMID- 21619717 TI - The dilemma of healthy eating and environmental sustainability: the case of fish. AB - OBJECTIVE: Despite widespread concern over exploitation of the European Union's fish stocks, dietary guidelines in the UK continue to recommend two portions of fish per week. The present study sought to investigate whether health and/or sustainability are motivating factors when purchasing and consuming fish and whether there are sociodemographic trends. DESIGN: A structured, self-completion postal questionnaire exploring consumers' attitudes towards purchasing fish, their dietary intake, stated purchasing behaviour and sociodemographic information. SETTING: Nottinghamshire, UK. SUBJECTS: Adults from 842 households randomly selected from the electoral register. RESULTS: Over half of the participants (57.0 %) were aware of the health benefits of fish consumption and reported health as a primary motivator for purchasing fish; however, only 26.8 % actively sought to purchase fish from a sustainable source (e.g. Marine Stewardship Council (MSC) certified fish). Only 30.6 % of participants met current dietary recommendations for fish intake. Older respondents (>60 years of age) were more likely to report purchasing fish for health reasons and to buy MSC fish. Participants were significantly less likely to report MSC purchases if they agreed with the statement 'I am confused about which type of fish I should be eating to protect fish stocks' (P < 0.001). CONCLUSIONS: The number of consumers purchasing fish for health reasons was more than those seeking sustainably sourced fish; yet, they still failed to meet the recommended intake set by the Food Standards Agency. Dietary advice to the public to increase consumption of fish conflicts with the prevailing pressure on fish stocks. Clear advice should be communicated enabling consumers to meet nutritional needs while protecting fish stocks. PMID- 21619718 TI - Electrospun Poly(gamma-Benzyl-L-Glutamate) and Its Alkali-Treated Meshes: Their Water Wettability and Cell-Adhesion Potential. AB - The aim of this study was (1) to fabricate non-woven meshes from a biodegradable polymer, poly(gamma-benzylL-glutamate), by electrospinning and subsequent hydrolysis of the ester bond on the polymer side-chain in an aqueous solution of NaOH, creating surface carboxyl groups on the fibers, and (2) to determine the effect of hydrolysis time on water wettability and cellular behaviors, in order to perform a preliminary evaluation for use of this polymer as a wound dressing matrix. A non-woven mesh composed of fibers, with minimal formation of 'bead' structures, was produced by electrospinning from tetrahydrofuran solution under optimally controlled conditions. The surface wettability largely depended on the hydrolysis time: an increase in hydrolysis time significantly reduced the advancing water contact angle. Instantaneous spreading of water droplets occurred at long hydrolysis times. An increase in hydrolysis time decreased adhesion of endothelial cells, but increased cell spreading. Neither proliferation nor invasion into the mesh interior was observed. We conclude by discussing the use of partially hydrolyzed non-woven mesh as a promising burn dressing. PMID- 21619719 TI - Study on the sampling of methadone from exhaled breath. AB - This study aimed at develop and validate the procedure for collecting exhaled breath for drug testing. Patients receiving methadone maintenance treatment were recruited for the study. Methadone levels were measured using liquid chromatography- electrospray-tandem mass spectrometry. The sampling device was based on a 47-mm C(18) filter and used under pressure to aid flow through the filter. The mouth was rinsed before sampling, and the device was constructed to protect against any saliva contamination. Methadone was present in breath samples before and after the daily intake of methadone. The mean (+/- SD) pre-dose level was found to be 135 +/- 109 pg/min (n = 48, median 121). The exhaled methadone increased after dose intake. Saliva levels of methadone were high in comparison with exhaled breath levels. Saliva contamination was suspected in about 10% of the collected samples. Similar results were obtained using 1, 3, and 10 min sampling times. The inter- and intraindividual variability were found to be similar and in the order of 50%. Alternative sampling using XAD-2 beads and solid phase microextraction fiber was found to be possible and enables sampling with low back pressure and with no need for pump assistance. The presented results confirm that breath testing is a new possibility for the detection of drugs of abuse. PMID- 21619720 TI - Ethyl glucuronide, ethyl sulfate, and ethanol in urine after intensive exposure to high ethanol content mouthwash. AB - To determine the degree of ethanol absorption and the resultant formation and urinary excretion of its conjugated metabolites following intensive use of high ethanol content mouthwash, 10 subjects gargled with Listerine((r)) antiseptic 4 times daily for 31/4 days. First morning void urine specimens were collected on each of the four study days and post-gargle specimens were collected at 2, 4, and 6 h after the final gargle of the study. Urine ethanol, ethyl glucuronide (EtG), ethyl sulfate (EtS), and creatinine were measured. Ethanol was below the positive threshold of 20 mg/dL in all of the urine specimens. EtG was undetectable in all pre-study urine specimens, but two pre-study specimens had detectable EtS (6 and 82 ng/mL; 16 and 83 MUg/g creatinine). Only one specimen contained detectable EtG (173 ng/mL; 117 MUg/g creatinine). EtS was detected in the urine of seven study subjects, but was not detected in the single specimen that had detectable EtG. The maximum EtS concentrations were 104 ng/mL and 112 MUg/g creatinine (in different subjects). Three subjects produced a total of eight (non-baseline) urinary EtS concentrations above 50 ng/mL or 50 MUg/g creatinine and three EtS concentrations exceeding 100 ng/mL or 100 MUg/g creatinine. In patients being monitored for ethanol use by urinary EtG and EtS concentrations, currently accepted EtG and EtS cutoffs of 500 ng/mL are adequate to distinguish between ethanol consumption and four times daily use of high ethanol content mouthwash. PMID- 21619721 TI - Analysis of 3,4-methylenedioxymetamphetamine: whole blood versus dried blood spots. AB - Analysis of dried blood spots is an increasingly accepted method in therapeutic drug monitoring, whereas its application by analogy to forensic samples has not been studied in detail. Therefore, we investigated whether determination of 3,4 methylenedioxymethamphetamine (MDMA) and its main metabolite 3,4 methylendioxyamphetamine (MDA) from dried blood spots (DBS) is as reliable as that from whole blood specimens. Analysis was performed by liquid chromatography tandem mass spectrometry following liquid-liquid extraction of blood and corresponding DBS samples from 20 volunteers participating in a controlled driving experiment under the influence of MDMA. The assay was checked for carryover, ion suppression/enhancement, linearity of response, lower limits of detection (LLOD) and quantitation, extraction efficiency and the within-run and between-run assay imprecision for both whole blood and DBS. The LLODs were 2.0 and 1.6 ng/mL for MDMA in whole blood and DBS, respectively, using a volume of 100 MUL. LLODs of MDA were determined to be 0.25 ng/mL in whole blood specimens and 0.12 ng/mL in DBS. Extraction efficiency and imprecision did not differ significantly between the two methods for both MDMA and MDA. The mean concentration ratio of corresponding whole blood and DBS samples, t-test, and the Bland-Altman difference plot were used to test hypothesis of equality. Statistical analyses revealed that methods did not significantly differ for MDMA or MDA. Thus, DBS analysis has potential as a precise and inexpensive alternative to whole blood analysis of MDMA. PMID- 21619722 TI - Detection of paraquat in oral fluid, plasma, and urine by capillary electrophoresis for diagnosis of acute poisoning. AB - The aim of this work was to develop a simple and fast method for the detection of paraquat in oral fluid, plasma, and urine by capillary electrophoresis with diode array detection (CE-DAD), to diagnose of acute poisoning related to this herbicide. The use of oral fluid in analytical toxicology has been established for drugs of abuse, but not for diagnosis of pesticides poisoning. Oral fluid was collected without stimulation using absorbent cotton swabs. Sample preparation included diluting, vortex mixing, and centrifuging of biological fluid, using ethyl paraquat as internal standard. CE-DAD analyses were performed in a fused silica capillary, and separation was performed under constant voltage condition of 21 kV, with detection at 195 nm. The electrolyte was a 40 mmol/L phosphate buffer at pH 2.50. The proposed method provided a fast and simple assay for the determination of paraquat in human oral fluid, plasma, and urine. To our knowledge, this is the first mention of use of oral fluid as a biological fluid diagnosis of quaternary ammonium herbicide poisoning. After validation, the method was applied to two cases of acute poisoning by this herbicide. PMID- 21619723 TI - Drug screening of whole blood by ultra-performance liquid chromatography-tandem mass spectrometry. AB - An ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS-MS) method for screening of drugs in whole blood has been developed and validated. Samples were prepared by supported liquid-liquid extraction on ChemElute((r)) columns with ethyl acetate/heptane (4:1). LC separation was achieved with an Acquity HSS T3-column (2.1 100 mm, 1.8-MUm particle). Mass detection was performed by positive ion mode electrospray MS-MS and included the following drugs/metabolites: morphine, codeine, ethyl morphine, oxycodone, buprenorphine, methadone, cocaine, methylphenidate, amphetamine, methamphetamine, 3,4 methylenedioxymethamphetamine (MDMA), Delta(9)-tetrahydrocannabinol (THC), fentanyl, alprazolam, bromazepam, clonazepam, diazepam, nordiazepam, 3-OH diazepam, fenazepam, flunitrazepam, lorazepam, nitrazepam, oxazepam, zopiclone, zolpidem, carisoprodol, and meprobamate. The cycle time was 9 min, and within- and between-day relative coefficients of variation varied from 1% to 33% and 2% to 58%, respectively. Extraction recoveries from whole blood were > 50% except for morphine and THC. The limit of quantitation was 0.1 to 521 ng/mL, depending on the drug. PMID- 21619724 TI - Zolpidem urine excretion profiles and cross-reactivity with ELISA((r)) kits in subjects using Zolpidem or Ambien((r)) CR as a prescription sleep aid. AB - Zolpidem, a Schedule IV controlled substance under the Federal Controlled Substance Act, has a rapid onset of action and short elimination half-life, rendering it ideal as a sleep aid. The crossreactivity of two zolpidem ELISA kits was investigated using patients taking a known administration of zolpidem. Subjects provided urine samples before, 30 min after their prescribed dose, and upon waking. Specimens were screened for zolpidem by ELISA (Immunalysis and Neogen) and then confirmed and quantitated for zolpidem using gas chromatography mass spectrometry (GC-MS) confirmation in select ion monitoring mode. All samples were measured for creatinine and corrected accordingly. The ELISA screening results demonstrated that all samples, except one, screened positive by ELISA using both kits, even when the GC-MS data found no zolpidem in the patient's urine sample. The maximum concentrations of zolpidem ranged from 15 to 120 ng/mg creatinine. Two of the patients showed zolpidem concentrations of 10 ng/mg creatinine or above after 20 h post dose. The high variability and concentration range seen in these patients, all on similar doses, suggest wide variability in the metabolism of zolpidem. PMID- 21619725 TI - Pharmacokinetic properties of succinylmonocholine in surgical patients. AB - Intoxications with succinylcholine (SUX) lead to a potentially lethal respiratory paralysis, and forensic cases involving accidental or deliberate SUX-application have been reported. Detection of SUX as well as its metabolite succinylmonocholine (SMC) is difficult: both substances are analytically challenging, and the extremely short plasma half-life of SUX additionally hampers detection of the parent compound. Pharmacokinetic data are scarce on SUX and non existent on SMC. To enhance forensic knowledge concerning SUX intoxications, plasma pharmacokinetics of SMC were investigated in anesthetized patients. Fifteen subjects scheduled for a surgical procedure were included in this study. Muscle-relaxation was initialized with a bolus injection of 80-100 mg SUX. Blood sampling was performed within 6 h after SUX application using paraoxonized tubes. Collected plasma was processed according to a validated isotope dilution high performance liquid chromatography-tandem mass spectrometry method using ion-pair solid-phase extraction. Pharmacokinetic parameters were derived from a user defined as well as a three-compartment model. For SMC, peak plasma concentrations were reached after 0.03-2.0 min. In contrast to SUX, SMC was more slowly and more extensively distributed, featuring triphasic plasma concentration time profiles. Pharmacokinetic key parameters were subject to interindividual variation of potential forensic importance, with terminal half-lives of 1-3 h indicating a detection interval of 8-24 h for SMC in plasma. SMC was proven to be the only realistic SUX marker in a forensic context. The present work defines meaningful detection windows for plasmatic SMC after SUX application and offers guideline values for forensic toxicological casework. PMID- 21619726 TI - Determination of illegally abused sedative-hypnotics in hair samples from drug offenders. AB - As several sedative-hypnotics are distributed illegally and are available domestically through media like the internet, their abuse is becoming a serious social problem. In the present study, four legal cases involving abuse of diazepam, midazolam, and/or zolpidem were proved by hair analysis using a simultaneous quantification method for the determination of diazepam (and its metabolites), lorazepam, midazolam, and zolpidem, which are often illegally abused in Korea, in hair that was developed and validated. Drugs and metabolites in hair were extracted using methanol followed by solid-phase extraction. The extracts were derivatized with N-methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA) and analyzed using gas chromatography-mass spectrometry in selected ion monitoring mode. The validation parameters of the method, including selectivity, linearity, limits of detection and quantification (LOQ), recovery, intra- and interassay precision and accuracy, and processed sample stability, were satisfactory. Moreover, the developed method was successfully applied to actual cases. In case 1, which involved a pop singer who was detained for suspected drug abuse, the concentrations of diazepam and nordiazepam were 5.7 and 2.0 ng/mg in nonpigmented hair and 6.6 and 1.8 ng/mg in pigmented hair, respectively. In case 2, 0.4 ng/mg zolpidem was detected in hair from a drug abuser who purchased illegally through the internet, and 0.2 ng/mg midazolam was detected in hair from an illegal drug seller in case 3. In case 4, diazepam (lower than the LOQ), nordiazepam (0.7 ng/mg), and zolpidem (0.7 ng/mg) were detected in hair from a medical doctor who abused drugs using forged prescriptions. PMID- 21619727 TI - Fatality from drinking denatured alcohol and hypothermia. AB - A 19-year-old non-diabetic female suffering from irritable bowel syndrome was found unconscious outdoors in the month of October. She was severely hypothermic and rushed to hospital for life-saving treatment. Evidence emerged that the victim had attempted suicide by drinking denatured alcohol (T-Red). According to the manufacturer of this product, it contains > 85% (v/v) ethanol, ~5% (v/v) acetone, 1-2% (v/v) ethyl acetate, and ~3% (v/v) methyl ethyl ketone (MEK), but no isopropanol. A venous blood sample taken on admission to hospital contained ethanol (660 mg/100 mL), acetone (25 mg/100 mL), isopropanol (78 mg/100 mL), and MEK, although the latter was not quantified. Despite intensive care, the patient died 21 h after admission and postmortem femoral blood contained ethanol (390 mg/100 mL), acetone (14 mg/100 mL), isopropanol (53 mg/100 mL), and MEK. During oxidative metabolism of ethanol, there is a shift in the redox state of the liver to a more reduced potential as reflected in a raised NADH/NAD(+) ratio, which impacts on other NAD-dependent biochemical reactions, including reduction of acetone to isopropanol. The lower concentrations of ethanol, acetone, and isopropanol in postmortem blood compared with antemortem blood indicate the metabolism of these substances during the 21-h survival period when the patient received emergency hospital treatment. PMID- 21619728 TI - Hydrogels Containing Core Cross-Linked Block Co-Polymer Micelles. AB - Poly(2-hydroxyethyl methacrylate) (pHEMA) hydrogels loaded with core cross-linked PEG-b-PCL micelles with different morphologies (spherical and rod-like) were prepared and evaluated for use as drugeluting soft contact lenses. The relationship between the composition of micelle-loaded pHEMA hydrogels and properties such as transparency and swelling were determined. The incorporation of core crosslinked micelles into pHEMA hydrogels led to the formation of different internal nanostructures which were dependent on the amount and morphology of the micelles added. 7-Hydroxy-9H-(1,3-dichloro-9,9'-dimethylacridin 2-one) (DDAO), a hydrophobic fluorescent dye, was loaded into the micelles prior to their incorporation within the hydrogel matrix. The in vitro release of DDAO demonstrated the potential of the micelles/pHEMA hydrogels to provide controlled drug delivery for at least 14 days. This study demonstrates the feasibility of both chemical and physical incorporation of block co-polymer micelles within pHEMA hydrogels as a means to achieve sustained release of drugs for potential application in ophthalmic therapies. PMID- 21619729 TI - Synthesis, Characteristics and Potential Application of Poly(beta-Amino Ester Urethane)-Based Multiblock Co-Polymers as an Injectable, Biodegradable and pH/Temperature-Sensitive Hydrogel System. AB - Physical polymeric hydrogels have significant potential for use as injectable depot drug/protein-delivery systems. In this study, a series of novel injectable, biodegradable and pH/temperature-sensitive multiblock co-polymer physical hydrogels composed of poly(ethylene glycol) (PEG) and poly(beta-amino ester urethane) (PEU) was synthesized by the polyaddition between the isocyanate groups of 1,6-diisocyanato hexamethylene and the hydroxyl groups of PEG and a synthesized monomer BTB (or ETE) in chloroform in the presence of dibutyltin dilaurate as a catalyst. The synthesized co-polymers were characterized by nuclear magnetic resonance spectroscopy, Fourier transform infrared spectroscopy and gel-permeation chromatography. Aqueous solutions of the co-polymers showed a sol-to-gel phase transition with increasing pH and a gel-to-sol phase transition with increasing temperature. The gel regions covered the physiological conditions (37 degrees C, pH 7.4) and could be controlled by changing the molecular weight of PEG, PEG/PEU ratio and co-polymer solution concentration. A gel formed rapidly in situ after injecting the co-polymer solution subcutaneously into SD rats and remained for more than 2 weeks in the body. The cytotoxicity tests confirmed the non-cytotoxicity of this co-polymer hydrogel. The controlled in vitro release of the model anticancer drug, doxorubicin, from this hydrogel occurred over a 7-day period. This hydrogel is a potential candidate for biomedical applications and drug/protein-delivery systems. PMID- 21619730 TI - Asymmetric composite membranes from chitosan and tricalcium phosphate useful for guided bone regeneration. AB - To fulfill the properties of barrier membranes useful for guided bone tissue regeneration in the treatment of periodontitis, in this study a simple process combining lyophilization with preheating treatment to produce asymmetric barrier membranes from biodegradable chitosan (CS) and functional beta-tricalcium phosphate (TCP) was proposed. By preheating TCP/CS (3:10, w/w) in an acetic acid solution at 40 degrees C, a skin layer that could greatly increase the mechanical properties of the membrane was formed. The asymmetric membrane with a skin layer had a modulus value almost 4-times that of the symmetric porous membrane produced only by lyophilization. This is beneficial for maintaining a secluded space for the bone regeneration, as well as to prevent the invasion of other tissues. The subsequent lyophilization at -20 degrees C then gave the rest of material an interconnected pore structure with high porosity (83.9-90.6%) and suitable pore size (50-150 MUm) which could promote the permeability and adhesiveness to bone cells, as demonstrated by the in vitro cell-culture of hFOB1.19 osteoblasts. Furthermore, the TCP particles added to CS could further increase the rigidity and the cell attachment and proliferation of hFOB1.19. The TCP/CS asymmetric composite membrane thus has the potential to be used as the barrier membrane for guided bone regeneration. PMID- 21619731 TI - Characterization of the physical properties and biocompatibility of polybenzoxazine-based aerogels for use as a novel hard-tissue scaffold. AB - The process to successfully synthesize polybenzoxazine (PBO)-based aerogels has recently been optimized; however, the biocompatibility of these materials has never been investigated. PBO is synthesized from bisphenol A and aniline, which are both precursors to many commonly used biomaterials, including polyurethane. Surface-wise these new aerogels resemble the innate extracellular matrix of bone and if these new aerogels exhibit acceptable biocompatibility, they may be used as a scaffold for bone tissue engineering. Here, we aimed to characterize some of the physical properties of PBO aerogels, PBO aerogels co-polymerized with resorcinol and formaldehyde (RF) and their conversion to carbon aerogel, while determining the compatibility of all of these materials towards human osteoblasts. Biocompatibility was determined with a live/dead cell cytotoxicity assay, a metabolic activity assay, alkaline phosphatase activity and osteocalcin production, after incubation with PBO-based aerogels for up to 5 days. PBO aerogels co-polymerized with RF tended to have a low density, porosity and elastic modulus and provided the weakest substrate for bone cell growth. PBO derived carbon aerogels tended to have a high density, a large porosity and improved mechanical properties and provided the best substrate for bone cell growth. These results suggest that PBO based carbon aerogels have a suitable biocompatibility towards osteoblasts and that they may be able to be used for bone tissue engineering scaffolds. PMID- 21619732 TI - Distribution of inflammatory cells, adhesion molecules, intermediate filaments, and chemokine receptors in subgroups of nasal polyp patients. AB - BACKGROUND: The pathogenesis of nasal polyps (NPs) is incompletely understood. The aim of this study was to investigate the distribution of inflammatory cells, adhesion molecules, intermediate filaments, and chemokine receptors in subgroups of NP patients. METHODS: In total, 35 patients were enrolled (group 1, 10 patients with Samter syndrome; group 2, 10 patients with diffuse polyposis without signs of Samter syndrome; group 3: 5 patients with solitary nasal polyps; group 4, 10 controls). Immunohistochemical staining was performed for CD105, CD106, CD62E, CD4, CD8, CXCR4, CD147, CD90, CD104, BF45, vimentin, pancytokeratin, and muscle-specific actin (MSA) in all patients' specimens. RESULTS: Expression of CD4, CD8, and CD106 were similar between the groups. Number of patients expressing CD4 in groups 1, 2, and 3 were higher than the controls. Number of patients expressing CD8 antigen were significantly higher in all three groups than in the control group. Expression of CD147 in groups 3 and 4 was significantly higher than in groups 1 and 2. CD98 expression was higher in groups 1, 2, and 3 than in group 4. The number of patients expressing vimentin in groups 1, 2, and 3 was significantly higher than in group 4. Immunostaining for pancytokeratin was positive in all patients. CONCLUSION: In conclusion, inflammatory cell, adhesion molecule, intermediate filament, and chemokine receptor profiles in nasal polyps differ among different patient groups and control subjects. Additional specific immunohistochemical studies are necessary for development of more specific immunotherapies. PMID- 21619734 TI - Report of the Global Oral Health Inequalities Task Group on periodontal disease: implications for dental hygienists. PMID- 21619735 TI - Prevention of root caries. PMID- 21619736 TI - Development and status of the advanced dental hygiene practitioner. AB - PURPOSE: Advanced dental hygiene practitioners (ADHPs), members of the oral health care team, bring care to persons disenfranchised from adequate dental services. ADHPs are licensed and provide the traditional educational, preventive and therapeutic dental hygiene services, plus diagnostic, prescriptive and minimally invasive restorative services. ADHPs work in collaboration with all members of the dental team, referring patients in need of services outside of their scope directly to dentists or other health care providers. PMID- 21619737 TI - Methadone and oral health--a brief review. AB - PURPOSE: Methadone is a prescription drug used to help individuals overcome withdraws from highly addictive illicit substances, such as heroin, but it has detrimental oral health effects. This manuscript reviews the oral health manifestations of methadone maintenance therapy, and considers its implications to oral care. It hopes to raise awareness among health care professionals, regulating bodies and the population at large about the oral side effects of methadone, the implications for dental treatment and considerations to better enhance the oral health of methadone users. The role of professional teams, particularly dentists and dental hygienists, is illustrated. PMID- 21619738 TI - Incorporating oral-systemic evidence into patient care: practice behaviors and barriers of North Carolina dental hygienists. AB - PURPOSE: Current research has reported associations between periodontal and systemic health, however, there is little data regarding how dental hygienists are incorporating this evidence into the dental hygiene practice. The purpose of this survey research was to determine what practice behaviors are prevalent among North Carolina dental hygienists regarding the incorporation of oral-systemic evidence into practice as well as perceived barriers to implementation. METHODS: A questionnaire was developed, pilot tested, revised and mailed to 1,665 licensed dental hygienists in North Carolina. After 3 mailings the response rate was 62%, with 52% (n=859) of respondents meeting inclusion criteria. Survey data was analyzed using descriptive statistics and Chi-square analysis. RESULTS: Respondents were predominately female (99%) with a 2 year degree (84%). While a minority of dental hygienists (20%) reported measuring blood pressure routinely on all patients, a majority (62%) measure blood pressure in select patients. Oral cancer screenings were performed by 89% of respondents. Eight percent record blood sugar levels, but only 3% record HbA1c values. Fifty percent of dental hygienists are extremely likely to refer patients to a medical provider for follow up assessments. Conditions dental hygienists are likely to discuss with patients include tobacco use (89%), pregnancy (84%) and genetics (79%). Significant barriers to implementing oral-systemic evidence include lack of time (52%), concern over legal risks (44%) and lack of education (27%). CONCLUSION: North Carolina dental hygienists are implementing some aspects of oral-systemic evidence into practice, but could take a more active role if they had more allotted time, education and training. PMID- 21619739 TI - Assessment of the skills and education necessary for a baccalaureate- prepared dental hygienist to pursue an entry-level role in clinical research. AB - PURPOSE: To assess the skills and education perceived as necessary for a baccalaureate-prepared dental hygienist to pursue an entry-level role in clinical research. METHODS: An electronic survey was developed and distributed to 124 dental hygienists. Participants held at least a baccalaureate level of education and were currently involved in clinical research or had previous clinical research experience. RESULTS: The survey response rate was 45% (n=56). Of the 56 respondents, 71% (n=40) met all inclusion criteria. The majority of respondents agreed that the University of Michigan Degree Completion and the Society of Clinical Research Associates pro gram competencies align with the skills and education needed to pursue an entry-level role in clinical research. Grant writing skills and the ability to prepare a manuscript for submission to a peer reviewed journal were not perceived as necessary for an entry-level position. CONCLUSION: Clinical research is a viable career option for dental hygienists. Obtaining a baccalaureate level of education will assist with acquiring entry level clinical research skills. Additional education is necessary to expand clinical research opportunities. Both education and mentoring are integral components for pursuing a career in clinical research. Expanding upon the research-related competencies of dental hygiene program curricula is one avenue for achieving these recommendations. PMID- 21619740 TI - Nurse practitioner's and certified nurse midwives' knowledge, opinions and practice behaviors regarding periodontal disease and adverse pregnancy outcomes. AB - PURPOSE: The purpose of this study was to assess the knowledge, opinions and practice behaviors of nurse practitioners (NP) and certified nurse midwives (CNM) regarding periodontal disease and adverse pregnancy outcomes. METHODS: A 45 item survey was developed, approved, pretested, revised and mailed to 404 North Carolina NPs and CNMs who provide prenatal care. Data was entered into an excel database and transferred to SPSS for Windows for complete analysis. Linear regression modeling was used to determine statistical significance. RESULTS: A total of 219 NPs and CNMs responded to the mailed survey, achieving a response rate of 54%. NPs and CNMs reported having limited knowledge regarding oral health. The majority felt they should collaborate with oral health care professionals to screen patients for periodontal disease. Most agreed they needed more information about periodontal disease and adverse pregnancy outcomes. CONCLUSION: NPs and CNMs who frequently examine women could serve an important role in screening for oral health problems and making appropriate dental health referrals. Increased basic and continuing education could prepare these professionals for collaborative care with oral health care professionals. This study suggests that collaboration between NPs and CNMs with dental professionals could lead to improved oral health care for pregnant patients. PMID- 21619741 TI - Interface with a community feeding team to address oral health of special needs children: a pilot project. AB - PURPOSE: Children with special health care needs (CSHCN) are most in need of anticipatory guidance and prevention. Achieving and maintaining optimal oral health is challenging, due to the many challenges this group faces both in medical and dental care. The purpose of this pilot project was to identify the educational needs of health professionals on a feeding team to prepare them to provide anticipatory guidance to special needs children, along with preliminary investigation into the role of the dental hygienist in improving the oral health of the CSHCN served by the feeding team. METHODS: Small focus groups were used to determine educational needs of the feeding team and provide initial identification of the role of the dental hygienist on the feeding team. RESULTS: The needs assessment indicated interest in an in-service to address connections between oral health and feeding issues, i.e.problems related to tube feeding and oral hypo- and hypersensitivities of the CSHCN. CONCLUSION: This project suggests there is a role for the dental hygienist on the feeding team to provide preventive dental care and referral as well as education for caregivers and therapists. Future research is needed to further delineate the role of the dental hygienist on the feeding team as well as implementation of a model for integrating them into this multidisciplinary team. PMID- 21619742 TI - Inter-examiner reliability of salivary diagnostic tests in a practice-based research network. AB - PURPOSE: Patient salivary characteristics are being measured as part of the Northwest PRECEDENT (Practice-based REsearch Collaborative in Evidence-based DENTistry) study on caries risk assessment. Prior to the implementation of these salivary diagnostic tests in a practice-based cohort study, inter-examiner reliability was assessed for resting salivary pH, stimulated salivary flow rate, pH and buffering capacity. METHODS: An initial evaluation of inter-examiner reliability of the 4 salivary tests was conducted among 6 dental auxiliary examiners using a convenience sample of 40 dental students. An in complete block design was used to assign samples to examiners (3 examiners per sample and 20 samples per examiner). Inter-examiner reliability testing was conducted on a patient population representative of the practice-based network in 4 member practices. Two dental assistants per practice independently conducted the salivary tests on samples provided by a random selection of 20 to 25 patients. A separate analysis was performed for each study. For each test, an inter-examiner reliability index was computed. RESULTS: Results from two studies are reported. In the first, stimulated salivary flow rate demonstrated excellent inter-examiner reliability, and resting salivary pH showed high inter-examiner reliability, while buffering capacity and stimulated salivary pH had moderate and very low inter-examiner reliability, respectively. In the second, inter-examiner reliability was excellent for the stimulated salivary flow rate and the resting salivary pH. The inter-examiner reliability for the stimulated salivary pH was also high and the stimulated salivary buffering capacity test had moderate reliability. CONCLUSION: The small variance in stimulated salivary pH and buffering capacity in dental students may have artificially made the reliability appear low in the first attempt at inter-examiner reliability testing. In the second study, all 4 tests had an accept able performance. PMID- 21619743 TI - Raising oral health awareness among nephrology nurses. AB - PURPOSE: The complexity of care for patients with chronic kidney disease (CKD) reinforces the need for collaboration between health care providers. The purpose of this study was to raise oral health awareness by providing educational seminars to health care providers, specifically nephrology nurses. METHODS: Educational seminars entitled "Oral Health and Chronic Kidney Disease" were delivered by calibrated dental hygiene educators to nurses in 3 different regions in the United States. The nurse participants (n=106) were given randomly assigned pre- and post-tests, assessing their knowledge of oral health and its relation to CKD. RESULTS: Pre-tests revealed that nurses had little knowledge of oral health and its relation to CKD. Regardless of questionnaire order, a significant increase of knowledge was observed for both groups (p-value <= 0.015), increasing from 61 to 76% on average. CONCLUSION: Incorporating interdisciplinary education increases nurses' knowledge and may lead to greater quality of life outcomes and improved overall health in patients with CKD. PMID- 21619744 TI - Identifying suspected breast cancer: development and validation of a clinical prediction rule. AB - BACKGROUND: An evidence-based approach is needed to identify women with breast symptoms who are most likely to have breast cancer so that timely and appropriate referral can take place. AIM: To report the development and validation of a clinical prediction rule for the diagnosis of breast cancer. DESIGN AND SETTING: Cohort study with two prospective groups of women: those presenting to a symptomatic breast clinic (derivation cohort) and a separate cohort presenting to 11 general practices (validation cohort) in Tayside, Scotland. METHOD: Regression analysis was used to derive a clinical prediction rule from presenting symptoms, personal and family history, and clinical findings. Validation consisted of estimating the number of breast cancers predicted to occur compared with the actual number of observed breast cancers across deciles of risk. RESULTS: In the derivation cohort of 802 patients, 59 (7%) were diagnosed with breast cancer. Independent clinical predictors for breast cancer were: increasing age by year (adjusted odds ratio [AOR] 1.10, 95% confidence interval [CI] = 1.07 to 1.13); presence of a discrete lump (AOR 15.20, 95% CI = 4.88 to 47.34); breast thickening (AOR 7.64, 95% CI = 2.23 to 26.11); lymphadenopathy (AOR 3.63, 95% CI = 1.33 to 9.92); and lump >= 2 cm (AOR 5.41, 95% CI = 2.36 to 12.38). All eight patients with skin tethering had breast cancer. The regression model had good predictive power, identifying all five breast cancers in the validation cohort of 97 patients in the top two deciles of risk. CONCLUSION: The clinical prediction rule discriminates between patients at high risk of breast cancer from those at low risk, and can be implemented as an evidence-based recommendation to enhance appropriate referral from general practice to a symptomatic breast clinic. Ongoing validation in further populations is required. PMID- 21619745 TI - Patients' confidence in their GP before and after being diagnosed with cancer. AB - BACKGROUND: General practice plays an important role in the cancer care pathway. The initial diagnostic phase may be crucial for the relationship between the patient and the GP. AIM: The aim was to describe whether patients' confidence in their GP changed after a cancer diagnosis, and analyse whether the change in confidence was associated with doctor delay. DESIGN AND SETTING: Population-based cohort study with 1892 questionnaires sent to patients and their GPs in general practices in the former Aarhus County, Denmark. METHOD: Information on patients' confidence in their GP was obtained from the patient questionnaire. Information on doctor delay was obtained from the GPs and defined as a period of 14 days or more from the date of first symptom presentation to the GP until cancer-specific investigation was initiated. RESULTS: Before the cancer diagnosis, 88.4% of the patients had confidence in their GP, which decreased to 80.0% after the diagnosis (P<0.001); 15.8% of the patients who experienced no doctor delay reported a decrease in confidence after the cancer diagnosis, compared with 29.1% of the patients with a doctor delay (P<0.001). Patients presenting with alarm symptoms and experiencing doctor delay were 3.8 times more likely to lose confidence compared with those presenting with alarm symptoms who experienced no doctor delay (P = 0.048). CONCLUSION: The majority of the patients had high levels of confidence in their GP before, as well as after, the cancer diagnosis. Nevertheless, a substantial amount had low confidence in the GP, especially when experiencing doctor delay in the initial phase of the pathway. PMID- 21619746 TI - How frequently do young people with potential cancer symptoms present in primary care? AB - BACKGROUND: Although uncommon in teenagers and young adults, cancer is the leading cause of non-accidental death in those aged 15-24 years. A prolonged period to cancer diagnosis in this cohort is reported and thought to be a consequence of the rarity of cancer in this age group, together with the complexity of presenting symptoms. Although diagnostic delay is perceived to be a problem for teenagers and young adults with cancer, little research has focused on their use of primary care services. AIM: To determine how often teenagers and young adults consult, their reasons for doing so, and how often potential oncological symptoms ('alert' symptoms) appear. DESIGN AND SETTING: Retrospective audit of consultations over 1 year. Three general medical practices in Scotland. METHOD: Medical records were examined for 2326 teenagers and young adults. Date of birth, sex, and free-text relating to the consultation were recorded and coded according to an agreed coding system; symptoms of potential oncological significance were coded as alert symptoms. RESULTS: A total of 1659 teenagers and young adults (71.3% of registered patients) attended their GP at least once. Females attended more frequently than males (P < 0.001), and older females more frequently than younger females (P < 0.001). Males exhibited no association between consultation frequency and age. The main reasons for consultation were pregnancy/contraception (15.8%) and infection (15.7%). Alert symptoms were uncommon, (reported in 4.0% of all consultations; 276 alert symptoms in 179 patients), and were not associated with age or sex. The most common alert symptoms were unexplained pain (34.8%), unexplained fatigue (14.5%), and lumps (13.4%). Two benign tumours were detected. CONCLUSION: A high proportion of teenagers and young adults consult their GP. Alert symptoms are uncommon and generally occur in isolation. More research is required to confirm these findings in a larger cohort and to examine how GPs respond to such alert symptoms. PMID- 21619747 TI - The diagnostic value of symptoms for colorectal cancer in primary care: a systematic review. AB - BACKGROUND: Over 37,000 new colorectal cancers are diagnosed in the UK each year. Most present symptomatically to primary care. AIM: To conduct a systematic review of the diagnostic value of symptoms associated with colorectal cancer. DESIGN: Systematic review. METHOD: MEDLINE, Embase, Cochrane Library, and CINAHL were searched to February 2010, for diagnostic studies of symptomatic adult patients in primary care. Studies of asymptomatic patients, screening, referred populations, or patients with colorectal cancer recurrences, or with fewer than 100 participants were excluded. The target condition was colorectal cancer. Data were extracted to estimate the diagnostic performance of each symptom or pair of symptoms. Data were pooled in a meta-analysis. The quality of studies was assessed with the QUADAS tool. RESULTS: Twenty-three studies were included. Positive predictive values (PPVs) for rectal bleeding from 13 papers ranged from 2.2% to 16%, with a pooled estimate of 8.1% (95% confidence interval [CI] = 6.0% to 11%) in those aged >= 50 years. Pooled PPV estimates for other symptoms were: abdominal pain (three studies) 3.3% (95% CI = 0.7% to 16%); and anaemia (four studies) 9.7% (95% CI = 3.5% to 27%). For rectal bleeding accompanied by weight loss or change in bowel habit, pooled positive likelihood ratios (PLRs) were 1.9 (95% CI = 1.3 to 2.8) and 1.8 (95% CI = 1.3 to 2.5) respectively, suggesting higher risk when both symptoms were present. Conversely, the PLR was one or less for abdominal pain, diarrhoea, or constipation accompanying rectal bleeding. CONCLUSION: The findings suggest that investigation of rectal bleeding or anaemia in primary care patients is warranted, irrespective of whether other symptoms are present. The risks from other single symptoms are lower, though multiple symptoms also warrant investigation. PMID- 21619748 TI - Impact on antibiotic prescription of rapid antigen detection testing in acute pharyngitis in adults: a randomised clinical trial. AB - BACKGROUND: Acute pharyngitis is one of the most frequent reasons for a GP consultation, and in most cases an antibiotic is prescribed. AIM: To determine the impact of rapid antigen detection testing (RADT) to identify group A beta haemolytic streptococcus in acute pharyngitis on the utilisation of antibiotics and appropriateness of their use. DESIGN AND SETTING: Cluster randomised controlled trial in primary care centres in Catalonia, Spain. METHOD: Patients with acute pharyngitis aged 14 years or older with at least one Centor criterion (fever, tonsillar exudate, tender enlarged anterior cervical lymph nodes, or absence of cough) were recruited. Participant physicians were randomly assigned to one of two study arms: an intervention group (assigned to RADT) and a control group (following usual care, without RADT). RESULTS: Of the 557 adults enrolled, 543 could be evaluated for analysis (281 [51.7%] in the intervention group and 262 [48.3%] in the control group). GPs without access to RADT were more likely to prescribe antibiotics compared with those who performed rapid tests (64.1% versus 43.8%, P<0.001). The more Centor criteria the patients presented, the greater the number of antibiotics prescribed, regardless of whether RADT was available (P<0.001). Antibiotics were prescribed in 30.7% of the cases with negative RADT results. Inappropriate antibiotic prescription was observed in 226 cases (43%), and was significantly greater in the control than in the intervention group (60% versus 26.9%; P<0.001). CONCLUSION: Even though more than 30% of negative RADT results resulted in antibiotic prescribing, the study findings support the use of RADT in the consultation. This strategy has an important impact on reducing antibiotic prescription among adults with acute pharyngitis. PMID- 21619749 TI - GPs' views in five European countries of interventions to promote prudent antibiotic use. AB - BACKGROUND: A variety of interventions have been developed to promote a more prudent use of antibiotics by implementing clinical guidelines. It is not yet clear which are most acceptable and feasible for implementation across a wide range of contexts. Previous research has been confined mainly to examining views of individual interventions in a national context. AIM: To explore GPs' views and experiences of strategies to promote a more prudent use of antibiotics, across five countries. DESIGN AND SETTING: Qualitative study using thematic and framework analysis in general practices in Belgium, France, Poland, Spain, and the UK. METHOD: Fifty-two semi-structured interviews explored GPs' views and experiences of strategies aimed at promoting a more prudent use of antibiotics. Interviews were carried out in person or over the telephone, transcribed verbatim, and translated into English where necessary for analysis. RESULTS: Themes were remarkably consistent across the countries. GPs had a preference for interventions that allowed discussion and comparison with local colleagues, which helped them to identify how their practice could improve. Other popular components of interventions included the use of near-patient tests to reduce diagnostic uncertainty, and the involvement of other health professionals to increase their responsibility for prescribing. CONCLUSION: The study findings could be used to inform future interventions to improve their acceptability to GPs. Consistency in views across countries indicates the potential for development of an intervention that could be implemented on a European scale. PMID- 21619750 TI - Cardiovascular multimorbidity: the effect of ethnicity on prevalence and risk factor management. AB - BACKGROUND: Multimorbidity is common in primary care populations. Within cardiovascular disease, important differences in disease prevalence and risk factor management by ethnicity are recognised. AIM: To examine the population burden of cardiovascular multimorbidity and the management of modifiable risk factors by ethnicity. DESIGN AND SETTING: Cross-sectional study of general practices (148/151) in the east London primary care trusts of Tower Hamlets, City and Hackney, and Newham, with a total population size of 843 720. METHOD: Using MIQUEST, patient data were extracted from five cardiovascular registers. Logistic regression analysis was used to examine the risk of being multimorbid by ethnic group, and the control of risk factors by ethnicity and burden of cardiovascular multimorbidity. RESULTS: The crude prevalence of cardiovascular multimorbidity among patients with at least one cardiovascular condition was 34%. People of non white ethnicity are more likely to be multimorbid than groups of white ethnicity, with adjusted odds ratios of 2.04 (95% confidence interval [CI] = 1.94 to 2.15) for South Asians and 1.23 (95% CI = 1.18 to 1.29) for groups of black ethnicity. Achievement of targets for blood pressure, cholesterol, and glycated haemoglobin (HbA(1c)) was higher for patients who were multimorbid than unimorbid. For cholesterol and blood pressure, South Asian patients achieved better control than those of white and black ethnicity. For HbA(1c) levels, patients of white ethnicity had an advantage over other groups as the morbidity burden increased. CONCLUSION: The burden of multiple disease varies by ethnicity. Risk factor management improves with increasing levels of cardiovascular multimorbidity, but clinically important differences by ethnicity remain and contribute to health inequalities. PMID- 21619751 TI - Epilepsy mortality and risk factors for death in epilepsy: a population-based study. AB - BACKGROUND: Epilepsy is an important cause of amenable mortality but risk factors for death in epilepsy are not well understood. AIM: To evaluate trends in epilepsy mortality in a large population and identify risk factors for death in epilepsy. DESIGN AND SETTING: Nested case-control study in the UK, using data from the General Practice Research Database (GPRD) from 1993 to 2007. METHOD: Participants were included if they had ever been diagnosed with epilepsy and prescribed anticonvulsant drugs. Trends in all-cause mortality in persons with epilepsy in the GPRD were compared with death registrations with epilepsy as the underlying cause. A nested case-control study was implemented to compare participants with epilepsy who died with those who did not die. RESULTS: The prevalence of epilepsy increased from 9 per 1000 in 1993 to 12 per 1000 in 2007, and epilepsy deaths also increased in this period. In a nested case-control study, mortality was associated with: recorded alcohol problems (odds ratio [OR] 2.96, 95% confidence interval [CI] = 2.25 to 3.89, P<0.001); having collected the last anticonvulsant prescription 90-182 days previously (OR 1.83, CI = 1.66 to 2.03, P<0.001); having an injury in the previous year (OR 1.41, 95% CI = 1.30 to 1.53, P<0.001), and having been treated for depression (OR 1.39, 95% CI = 1.28 to 1.50, P<0.001). In data available from 2004 onwards, being recorded seizure free in the previous 12 months was associated with lower mortality (OR 0.78, 95% CI = 0.71 to 0.86, P<0.001). CONCLUSION: Mortality with epilepsy appears to be increasing. Patients who have alcohol problems, do not collect repeat prescriptions for anticonvulsant drugs, have recent injuries, or have been treated for depression may be at increased risk of death; patients who remain seizure free over 12 months are at a lower risk. PMID- 21619752 TI - Impact of the QOF and the NICE guideline in the diagnosis and management of depression: a qualitative study. AB - BACKGROUND: The National Institute for Health and clinical Excellence (NICE) depression guideline (2004) and the updated Quality and Outcomes Framework (QOF) (2006) in general practice have introduced the concepts of screening severity assessment, for example using the Patient Health Questionnaire 9 (PHQ-9), and 'stepped care' for depression. AIM: To explore primary care practitioner perspectives on the clinical utility of the NICE guideline and the impact of the QOF on diagnosis and management of depression in routine practice. DESIGN AND SETTING: Qualitative study using focus groups from four multidisciplinary practice teams with diverse populations in south Yorkshire. METHOD: Four focus groups were conducted, using a topic guide and audiotaping. There were 38 participants: GPs, nurses, doctors in training, mental health workers, and a manager. Data analysis was iterative and thematic. RESULTS: The NICE guideline, with its embedded principles of holism and evidence-based practice, was viewed positively but its impact was compromised by resource and practitioner barriers to implementation. The perceived imposition of the screening questions and severity assessments (PHQ-9) with no responsive training had required practitioners to work hard to minimise negative impacts on their work, for example: constantly adapting consultations to tick boxes; avoiding triggering open displays of distress without the time to offer appropriate care; positively managing how their patients were labelled. Further confusion was experienced around the evolving content of psychological interventions for depression. CONCLUSION: Organisational barriers to the implementation of the NICE guideline and the limited scope of the QOF highlight the need for policy makers to work more effectively with the complex realities of general practice in order to systematically improve the quality and delivery of 'managed' care for depression. PMID- 21619753 TI - Recording ethnicity in primary care: assessing the methods and impact. PMID- 21619755 TI - Improving cancer outcomes: better access to diagnostics in primary care could be critical. PMID- 21619756 TI - The future of national health systems. PMID- 21619757 TI - Research for commissioners: filling a black hole in the NHS White Paper. PMID- 21619758 TI - Questionnaire severity measures for depression. PMID- 21619759 TI - Trainees working in psychiatry. PMID- 21619760 TI - Revalidation. PMID- 21619762 TI - Doctors and depression. PMID- 21619761 TI - Novel treatments for type 2 diabetes. PMID- 21619763 TI - How to protect general practice from child protection. PMID- 21619764 TI - Oral and maxillofacial surgery. PMID- 21619765 TI - Ubi Scientia in the midst of the cosy cardigans of Caritas? PMID- 21619766 TI - 'Heartsink' patients in general practice: a defining paper, its impact, and psychodynamic potential. PMID- 21619767 TI - Second thoughts about the NHS reforms. PMID- 21619768 TI - Social prescribing in very deprived areas. PMID- 21619770 TI - Seeing with new eyes: what can general practice learn from the science of improvement? PMID- 21619771 TI - Pulse oximetry in primary care: primary care diagnostic technology update. PMID- 21619772 TI - Nocturnal enuresis in children and young people: NICE clinical guideline. PMID- 21619773 TI - Tips for GP trainees working in ENT. PMID- 21619774 TI - Emerging molecular prognostic markers in human thyroid carcinoma. PMID- 21619775 TI - Duplex ultrasound and magnetic resonance imaging of the supra-aortic arches in patients with non recurrent inferior laryngeal nerve: a comparative study. AB - BACKGROUND: Non-recurrent inferior laryngeal nerve (NRILN) is usually discovered during thyroid surgery. It is often associated with vascular abnormalities that can be detected with magnetic resonance imaging (MRI) or duplex ultrasound scan. The aim of this study was to compare the diagnostic sensitivity of ultrasonography with MRI to identify the vascular abnormalities associated to NRILN. PATIENTS AND METHODS: We revised 2713 total thyroidectomies to select patients with NRILN. The NRILN was identified in 17 patients (0,6%). A postoperative ultrasonic duplex scanning and a MRI was performed in 15 cases as 2 patients refused to submit to the exams. RESULTS: At MRI an unique origin of common carotid trunk and a concomitant aberrant retroesophageal subclavian right artery was showed in 11 patients. In 2 cases vascular abnormality consisted in separated origin of supra-aortic arteries. At duplex ultrasound scan only in 2 patients was impossible to identify vascular abnormalities detected at MRI. Tthe diagnostic sensitivity of duplex ultrasound was 84,6%. CONCLUSIONS: Preoperative duplex ultrasound is a non invasive method with high diagnostic sensitivity that can easily complete the preoperative thyroid ultrasonography. PMID- 21619776 TI - Non-recurrent laryngeal nerve coexisting with ipsilateral recurrent nerve: personal experience and literature review. AB - INTRODUCTION: Non-recurrence and variations in ascending course of the recurrent laryngeal nerve (RLN) represent a risk factor for nerve injuries during thyroid surgery. Non-recurrent laryngeal nerve (NRLN) coexisting to recurrent nerve branch is a rare anatomic anomaly. It could be a cause of nerve injuries during thyroidectomy. A systematic intraoperative nerve identification may allow an effectiveness prevention of iatrogenic injuries. CASE REPORT: We report one case of a young woman underwent to total thyroidectomy (TT) for papillary thyroid carcinoma (PTC) where we found a rare variation of the right inferior laryngeal nerve anatomy. We identified both right laryngeal nerve structures before completing thyroidectomy avoiding possible nerve damage. The postoperative course was without complications. DISCUSSION: Iatrogenic injury of RLN is one of the most serious complication in thyroid surgery. Several risk factors favouring this complication were found as the presence of anatomic variations of the inferior laryngeal nerve. Identification of a normal caliber recurrent nerve can allow the surgeon to complete the thyroid excision; diversely, in case of a smaller caliber nerve in the usual recurrent course, a careful dissection should be continued to demonstrate a possible merger with ipsilateral non-recurrent nerve. CONCLUSIONS: The aim of this paper is to report a rare case of NRLN associated to a smaller caliber branch of RNL. We emphasize that careful dissection and intimate knowledge of normal and anomaly anatomy allow for avoidance of nerve injury during surgery in the neck. PMID- 21619777 TI - Ectopic liver nodules: a rare finding during cholecystectomy. AB - The ectopic liver (or choristoma) is a rare condition found during autopsy or abdominal exploration for various indications. The authors report two cases of ectopic liver found during laparoscopic cholecystectomy for acute cholestytis. The ectopic liver tissue has been reported to develop in several sites as thoracic cavity, gastrohepatic ligament, adrenal glands, pancreas, esophagus and, above all, gallbladder. The Authors review the literature and report their experience as a contribution to the knowledge of this rare pathological entity. PMID- 21619778 TI - Heterotopic pancreatic tissue in the gallbladder. Two case reports and brief review of the literature. AB - Heterotopic pancreas tissue in the gallbladder is a rare benign condition with no clinical relevance and found incidentally in most cases. Only 28 cases of heterotopic pancreas are reported until 2007 in the worldwide literature. The preoperative diagnosis is very hard, so that it could be considered in differential diagnosis with other benign and malignant disease of gallbladder and biliary tree. The surgical approach for the excision of a undefined lesion and the histological identification is a correct procedure in suspect case. We report 2 cases of heterotopic pancreatic tissue of the gallbladder. PMID- 21619779 TI - Isolated inguinal endometriosis. Case report with ultrasonographic preoperative diagnosis. AB - Inguinal endometriosis is rare and accounts for 0.3-0.6% of patients affected by endometriosis. A correct preoperative diagnosis is rare. Diagnosis is frequently made by histologic examination. A 36-year-old nulliparous woman presented with a painful mass in her right groin of 2 years duration. The pain fluctuated according to the menstrual period. Physical examination revealed an elastic hard mobile mass measuring 2 x 2 cm in the right inguinal region. Ultrasound examination confirmed a hypoechoic tumor in the right inguinal region with poorly defined boundaries and perilesional and intralesional vascular flow suspect for endometriosis. Wide excision of the lump with a part of the round ligament was carried out. Histology showed endometrial glands and stroma within the fibrous tissue. The patient had an uneventful recovery and was discharged the next day. After surgery, the pain disappeared completely. No signs of recurrence occurred at approximately 16 months after the surgery. Although rare, extrapelvic endometriosis should be considered in the differential diagnosis in women of reproductive age presenting with an inguinal mass, especially if the groin mass is associated in size and tenderness with menstrual variability. US appearance is very useful in diagnosis so ultrasonography can be considered the examination of choice. PMID- 21619780 TI - Cortisone-induced gigantomastia during chemotherapy. AB - Gigantomastia is a rare, psychologically and physically disabling condition characterized by excessive breast growth. There is no universal consensus on the definition of gigantomastia, but it is most commonly described as breast enlargement that requires removal ranging from 800 to 2000 g, or even a D cup bra size. It typically occurs in the setting of marked hormonal changes such as puberty and pregnancy; however, there have also been a number of reports of gigantomastia in the setting of autoimmune diseases. Rare association of gigantomastia included medicinal aetiologies such as penicillamine, neothetazone, and cyclosporine. The mechanism of action of these pharmacological agents remains unclear. We report the first case of gigantomastia associated with cortisone in the setting of ovary cancer treated with chemotherapy cycles after hysterectomy and bilateral adnexectomy. Moreover, we propose a clinic evidence and a metabolic theory to explain this association. PMID- 21619781 TI - A wrong diagnosis of recurrent perineal hernia. AB - We describe a case of a patient who had several operations for recurrent perineal hernia. She eventually had an abdominal surgical debulking for aggressive angiomyxoma. PMID- 21619782 TI - Hemoperitoneum as severe and unusual complication in the stapler recto-anopexy for hemorrhoidal prolapse. Case report. AB - We report unusual but severe complication after Longo recto-anopexy for hemorrhoidal prolapse, i.e. large intramural hematoma of the rectum and subsequent hemoperitoneum. We make some assessment about the technique. PMID- 21619783 TI - Open and endovascular combined procedures in lower limb arterial reconstructions. AB - INTRODUCTION: The association of open and endovascular procedures in patients with obstructive disease of the lower limbs gives the opportunity to treat in a single step multi-segmental lesions which were currently treated in the past by more demanding open procedures. PATIENTS AND METHODS: From January 2003 to May 2009, 52 patients with peripheral occlusive chronic arterial disease of the lower limbs were submitted to combined open and endovascular procedures. In 37 patients (71.2%) a femoro-popliteal/pedidial bypass was carried out after Percutaneous Transublimal Arterio (PTA)/stenting of the iliac/femoral arteries, while in the remaining 15 patients (28.8%) the endovascular procedures were performed following open approach, that consisted in a femoro-popliteal bypass. Iliac stenting interested the common iliac artery in 19 patients (51.3%), the common and external iliac artery in 11 patients (29.7%) and the external artery in 7 (19%). In this same group a fem-pop bypass below knee was carried out in 28 (75.6), a fem-pop above knee in 6 (16.2%), a fem-pop and jump on the posterior tibial in 2 (5.4%) and a superficial femoral-pedal in the last one (2.8%). In the other group the fem-pop bypass below knee was followed by a PTA of the tibial vessel in 12 cases (80%) and by a stenting in only 3 cases (20%). RESULTS: Three patient was lost to follow-up. Three (8.1%) iliac stents became occluded and the sudden onset of an acute ischemia of the limb required in all cases an aorto femoral bypass. The occlusion of tibial stent in 2 patient (13.3%) was no clinical relevant, and the failure of the PTA procedure in other 4 patients with the reocclusion of the tibial arteries was followed by thight amputation. Occlusion of the femoro-popliteal bypass occurred in 9 of 37 patients (24.3%) submitted to iliac stenting and in 4 of 15 (26.6%) patients with PTA of the tibial vessels. The overall patency rate was 69.2%. Limb salvage was 77.5% CONCLUSIONS: Combination of the open and endovascular procedures to treat lesions of the lower limbs is effective and durable in terms of patency and complication rate. For validation this new approach needs the recruitment of much more patients and prospective protocol studies. PMID- 21619784 TI - Laparoscopic splenectomy in patients under the age of eighteen. Experience in 18 cases. AB - BACKGROUND: Pediatric surgery is now in the forefront of minimal access procedures. Although pediatric surgeons have been skeptic about laparoscopic splenectomy, recently minimally invasive approach for spleen removal has been revaluated also in young patients. Purpose of this study was to report Authors' personal experience in patients under eighteen who underwent laparoscopic splenectomy. Results of the procedure were evaluated. PATIENTS AND METHODS: A retrospective review was undertaken in 18 splenectomised patients under the age of eighteen. Indications were hereditary spherocytosis in 10 patients, beta thalassemia in 4, idiopathic thrombocytopenic purpura in 3 and a splenic cyst in 1 child. RESULTS: No intra-operative complications occurred. No conversion to open surgery was reported. During the follow-up one case of portal vein thrombosis, treated by medical therapy, was encountered and no other postoperative complications were observed. CONCLUSIONS: Laparoscopic approach has to be preferable for all those children undergoing spleen surgery. In experienced hands, it is of beneficial effects with a very reasonable rate of complications. PMID- 21619785 TI - [A long way to go for prevention and treatment of organic solvents poisoning]. PMID- 21619786 TI - [Induction of cell damage and change of miR-133b expression by paraquat in PC12 cells]. AB - OBJECTIVE: To investigate the effect of paraquat on induction of cell damage and miR-133b expression in PC12 cells. METHODS: Cytotoxicity of PC12 cells was measured by MTT assay, following the PC12 cells treatment with 50, 100, or 300 umol/L paraquat. Cell apoptosis was examined by the method of Annexin V-FITC/PI in flow cytometry (FCM) and the relative level of miR-133b expression was measured by real time RT-PCR, following the PC12 cells treatment with 100 or 300 umol/L paraquat. RESULTS: Survival rate of PC12 cells treated with 100 or 300 umol/L paraquat was lower than that of the vehicle control group (P < 0.01, P < 0.05), in the dose dependent pattern. Apoptotic rate of PC12 cells treated with 100, 300 umol/L paraquat was higher than that of the vehicle control group (P < 0.05). The relative level of miR-133b expression of PC12 cells treated with 300 umol/L paraquat was higher than that of the vehicle control group (P < 0.05). CONCLUSIONS: Paraquat may cause cell damage and induce apoptosis in PC12 cells, and induce miR-133b expression. PMID- 21619787 TI - [The influence of SiO2 on epithelial-mesenchymal transition (EMT) in human bronchial epithelial cells]. AB - OBJECTIVE: To investigate SiO2-induced EMT in human bronchial epithelial cells HBE in vitro. METHODS: HBE cells were cultured and then stimulated with indicated doses of SiO2 (0, 50, 100, 200, 300 ug/ml). The morphological changes were observed by microscope. In addition, Western blot was per-formed to detect the expression of E-cad, alpha-SMA and Vim. The changes of migration ability were examined by wound-healing assay in vitro. RESULTS: (1) After exposure to SiO2, HBE cells lost contact with their neighbor and displayed a spindle-shape, fibroblast-like morphology. (2) Compared with the control, the E-cad (300 ug/ml group) expression downregulated 2.98 fold (P < 0.05), and the Vim (300 ug/ml group) and alpha-SMA (200 ug/ml group) expression upregulated 4.46 fold and 3.55 fold (P < 0.05). There were significant differences between 100, 200, 300 ug/ml groups and the control group (P < 0.05). (3) In the test group, the percentage of wound-healing areas/wound areas were larger than those in control group (P < 0.05). CONCLUSIONS: SiO2 could induce EMT in human bronchial epithelial cells. PMID- 21619788 TI - [Modification of GSTM1, GSTT1 and GSTP1 gene polymorphisms on urinary 1 hydroxypyrene excretions]. AB - OBJECTIVE: To investigate the modification of GSTM1, GSTT1 and GSTP1 gene polymorphisms on urinary 1-hydroxypyrene (1-OHP) excretions in workers under different exposure levels. METHODS: Four hundred and forty-seven occupationally exposed workers from two coking plants and 220 control workers from a wire rod plant were genotyped to analyze the modification of GSTM1, GSTT1 and GSTP1 gene polymorphisms on urinary 1-OHP excretions. RESULTS: The urinary 1-OHP concentration in exposed group was much higher than that in control group (4.61 vs 0.34 umol/mol Cr, P < 0.05). Occupational exposure levels and cigarette smoking were of the dominating factors affecting 1-OHP excretions in urine. After controlling potential confounders, decreased excretion of urinary 1-OHP was associated with GSTP1 I105V AG + GG genotype in coke oven workers (single-gene model, P = 0.012; multi-gene model, P = 0.011) and with GSTT1 null type in the analysis including all subjects (P = 0.055 in both single-gene and multi-gene models). GSTT1 and GSTP1 were interacted on the urinary concentrations of 1-OHP. CONCLUSION: Urinary 1-OHP concentrations can be modified by GSTM1, GSTT1 and GSTP1 gene polymorphisms, indicating that these genes are involved in the metabolism of polycyclic aromatic hydrocarbons. PMID- 21619789 TI - [Detecting DNA damage of human lymphocytes exposed to 1,2-DCE with gammaH2AX identified antibody using flow cytometer assay]. AB - OBJECTIVE: To study DNA damage of human peripheral blood lymphocytes exposed to 1,2-dichloroethane (1,2-DCE) with flow cytometry (FCM) assay. METHODS: The lymphocytes were obtained from 21 workers who are occupationally exposed to 1,2 DCE (exposed group) and 27 workers who were not exposed to 1,2-DCE in the same factory (inner control) and 28 island residents who had never been occupationally exposed to adverse factors (external control). FCM assay was adopted to detect DNA damage of the lymphocytes of each group. Lymphocytes of the health people were incubated with 1,2-DCE at different doses, and FCM assay was used to detect DNA damage. RESULTS: DNA damage rate (%) of the exposed group of exposed workers (4.05% +/- 2.55%) was significantly higher than the inner control group of workers (1.97% +/- 1.40%) and external control groups of island residents (0.23% +/- 0.13%), and the DNA damage of inner control was higher than the external control, all the differences were statistically significant (P < 0.01 or P < 0.05). The geometric mean fluorescence intensity of the workers in the exposed group (3.33 +/- 3.01) was significantly higher than the (2.07 +/- 0.58) only (P < 0.05). There was no significant difference in the DNA damage rate as well as the geometric mean fluorescence intensity among the exposed group of workers with different years of working period (P > 0.05). In vitro, the fluorescence intensity at the dose of 20, 30 umol/L for 0.5 h exposure showed statistical significance compared with the negative control group (P < 0.01). The DNA damage rate at the dose of 20, 30 umol/L for 1.0 h exposure was statistically significant compared with the negative control group (P < 0.05, P < 0.01); The fluorescence intensity at the dose of 10, 20, 30 umol/L for 1.0 h exposure was statistically significant compared with the negative control group (P < 0.05, P < 0.01). CONCLUSION: 1,2-DEC can cause DNA damage. And gammaH2AX FCM assay can be a sensitive, objective and effective method of detecting DNA damage of peripheral blood lymphocytes. PMID- 21619790 TI - [Influence of MDR1 gene C3435T on peripheral white blood cell counts in workers exposed to benzene]. AB - OBJECTIVE: To explore the effects of MDR1 C3435T on the peripheral white blood cell counts in workers exposed to benzene. METHODS: One hundred and twenty-one benzene-exposed workers and 110 healthy controls without benzene exposure were enrolled in this study. White blood cell counts influenced by the polymorphism of MDR1 gene were analyzed. RESULTS: The frequency of MDR1 3435 C/C, C/T, T/T in healthy controls was 37.27%, 46.36%, 16.37%, respectively, and it was 38.84%, 41.33%, 19.83% in the benzene-exposed workers, respectively. The frequency of the MDR1 gene was also not significantly different between benzene exposed workers and controls. Subjects exposed to benzene with MDR1 3435 mutation genotype (T/T) had the significantly lower WBC [(5.46 +/- 1.51) * 10(9)/L] than those carrying wild type (C/C) and heterozygous (C/T), whose WBC were (6.08 +/- 1.28) * 10(9)/L (P = 0.044). CONCLUSION: P-glycoprotein encoded by MDR1 gene may be implicated into the hematotoxicity of benzene. Subjects carrying MDR1 3435 T/T genotype may have a higher risk of benzene poisoning. PMID- 21619791 TI - [Establishment of acute pulmonary edema model induced by high concentrations of nitrogen dioxide in rats]. AB - OBJECTIVE: To establish the rats model of acute pulmonary edema induced by inhalation of high concentrations of nitrogen dioxide (NO2). METHODS: 38 SD rats were divided into the experimental group (n = 30) and the control group (n = 8). 30 rats in the experimental group were exposed to (6747.47 +/- 25.24) mg/m(3) NO2 in the exposure system. At the time point of 6, 12, 18, 24 h, chest X-ray examination was taken for the experimental group. And at each time point, 6 rats were sacrificed after taking blood samples. After sacrificing, the lung of rats was taken for pathological examination and calculated lung wet/dry weight ratio. Erythrocyte superoxide dismutase (SOD) activity and plasma atrial natriuretic peptide (ANP) concentration of blood samples were detected. RESULTS: Acute pulmonary edema was successfully induced by exposure to NO2 in 30 rats within 24 hours. There were some cloudy shadows without clear edge on the chest X-ray. To the time point of 12 hours, shadows combined with each other, and to the time point of 18 hours, the whole lung became "white" on the X-ray. The situation stabilized but not improved at the time point of 24 hours. HE staining of the lung tissue showed that to the time point of 6 hours, the alveolar gap increased and small amount of eosinophilic liquid leaked into alveolar. To the time point of 12 hours, alveolar combined with each other and eosinophilic liquid increased in amount. To the time point of 18 hours, the whole alveolar was filled with eosinophilic liquid and the situation stabilized till the time point of 24 hours. Wet/dry weight ratio of the experimental group at each time point were 5.6 +/- 0.20, 6.89 +/- 0.25, 8.03 +/- 0.47, 7.81 +/- 0.45. There was significant difference compared with the control group which was 4.72 +/- 0.06 (P < 0.01). There was statistical difference between 12, 18, 24 h and 6 h time points (P < 0.01). Moreover, statistical difference was observed between 18, 24 h and 12 h time points for wet/dry weight ratio (P < 0.01). The erythrocyte SOD activity reduced significantly. Compared with the control group, there was a statistical difference (P < 0.01) at each time point. After exposure of 18 and 24 hours, plasma ANP concentration (136.66 +/- 35.37) and (134.10 +/- 60.41) ng/ml respectively, which were higher than (31.31 +/- 13.06) ng/ml of control group and (34.71 +/- 13.42) ng/ml of 6 hours time point and (47.98 +/- 7.86) ng/ml. The differences were significant (P < 0.01). CONCLUSION: High concentrations of NO2 can induce acute pulmonary edema model successfully in SD rats. PMID- 21619792 TI - [mRNA expression and methylation status of p15 promoter in mouse bone marrow cells exposed to 1,4-benzoquinone]. AB - OBJECTIVE: To detect the expression and the CpG island methylation status of tumor suppressor gene p15 after exposure to 1,4-benzoquinone (1,4-BQ) in primary cultivated C57BL/6J mouse bone marrow cells in vitro. METHODS: The mouse bone marrow cells were isolated in vitro. The effect of 0, 0.1, 1, 5, 10, 20, and 40 umol/L 1,4-BQ on cell viability (CKK-8) was detected. 0, 0.1, 1, 10 umol/L 1,4-BQ were used to intoxicate the mouse bone marrow cells for 24 h; Real-time PCR was employed to analyze the mRNA expression level of p15; The bisulfite sequencing PCR (BSP) was used to look into the methylation status of CpG islands in p15 promoter region. RESULTS: 1,4-BQ exhibited dose-dependent toxicity to mouse bone marrow cells, and the LC(50) was 8.3 umol/L (95%CI: 4.6 - 10.6 umol/L). The mRNA expression of p15 in 10 umol/L group was only equivalent to 43% of control group. Compared with control group, the decrease of p15 mRNA expression in1 and 10 umol/L concentration were obvious, and the differences had statistical significance (P < 0.05 or P < 0.01). BSP sequencing results were same between the exposure groups and control group, the 56 CpG sites on CpG islands remained in the state of unmethylated. CONCLUSION: mRNA expression of p15 gene decreases after exposure to 1,4-BQ, but the CpG islands methylation status in promoter is not affected, suggesting that methylation does not participate in 1,4-BQ-mediated p15 gene expression decrease, other effect mechanisms still need to be investigated. PMID- 21619793 TI - [An accident of acute hydrogen sulfide poisoning]. PMID- 21619794 TI - [Effects of co-exposure to paraquat and maneb on system of substantial nigra and striatum in rats]. AB - OBJECTIVE: To investigate the effects of exposure of paraquat and maneb on the behavior, the morphology and electrical activity of the Substantia nigra and striatum, and to discuss the relationship between this two pesticides and Parkinson's disease. METHODS: 37 rats were divided randomly into 3 groups: control group (n = 11), paraquat (10 mg/kg) group (n = 13) and combinative group of paraquat (10 mg/kg) and maneb (30 mg/kg) (n = 13), and were exposed twice a week for 6 weeks by intraperitoneal injection. The behavior of animals in the declined-plane, the vertical-grid and the open-field test were observed. The morphology of substantia nigral neurons were investigated by HE pathology. The spontaneous discharge of striatum neurons were recorded after exposure. RESULTS: Compared to the control group and the pre-exposure group, both the numbers of animals sliding down from the declined-plane and the latency of rats' moving on the vertical-grid significantly increased, and the animals' autonomic movement decreased significantly (P < 0.05, P < 0.001). After the combinative exposure, the neurons of the Substantial nigra pars compacta (SNPc) were progressively impaired, the cell density of the paraquat group [(82.17 +/- 12.91) n/mm(2)] and the combined group [(41.15 +/- 6.44) n/mm(2)] were lower than that in control group (143.10 +/- 20.85 n/mm(2)] (P < 0.01). In the paraquat group (5.97 +/- 7.30 Hz) and the combined group [(6.95 +/- 9.87) Hz], the average discharge rates of the striatum neurons were increased significantly compared to the control group [(1.78 +/- 5.05) Hz] (P < 0.01). The bursting discharge was increased significantly in the combined group (22.3%) compared to the control group (9.8%) and the paraquat group (5.6%) (P < 0.05, P < 0.01). CONCLUSION: The co-exposure of paraquat and maneb could induce similar symptoms to Parkinsonism syndrome of rats such as rigidity, moving reduction and etc, and the combined exposure had a certain enhanced effect compared to alone paraquat exposure. The combinative exposure of paraquat and maneb could cause neural loss in SNPc and it is involved with the enhanced electrophysiological activity in striatum. The synergy toxicity of paraquat and maneb in nigrostriatal system is related to Parkinson's disease. PMID- 21619795 TI - [An accident of chloroethyl chloroformate poisoning]. PMID- 21619796 TI - [Hepatocyte growth factor combined with autologous bone marrow mesenchymal stem cell transplantation for treatment of silicosis]. AB - OBJECTIVE: To evaluate the potential role of hepatocyte growth factor (HGF) combined with bone marrow mesenchymal stem cells (BMSC) autograft for the treatment of silicosis. METHODS: Bone marrow (100 ml) was aspirated from a severe silicosis patient. BMSCs isolated, purified and cultured in vitro. When BMSC came to 70% confluence at passage 3, the culture medium was added liposomes (lipo2000) and plasmid-HGF (p-HGF) and cultured for 2 d. HGF-MSCSs (5 * 10(7) cells) were resuspended in 50 ml 0.9% sodium chloride (NS) and infused Intravenous drip at 3 consecutive times (once a week). Clinical follow-up were performed before and after treatment: (1) pulmonary high-kV X-ray, chest CT examination; (2) pulmonary function test; (3) determination of serum ceruloplasmin. RESULTS: The symptoms such as coughing, chest tightness disappeared at 12 months after treatment. Pulmonary function tests showed significant changes after treatment: forced vital capacity (FVC) increased from 64.6% to 81.0%, forced expiratory volume in one second (FEV(1.0)) increased from 68.7% to 90.1%, 1 second rate (FEV(1.0)/FVC%) reduced from 111.6% to 107.1%, the maximum mid-expiratory flow (FEF(25%~75%) decreased from 100.2% to 94.6%, forced expiratory vital capacity 75% of the moment bit of gas flow (MEF(75%)) increased from 99.2% to 113.5%, forced expiratory vital capacity 50% of the moment bit of gas flow (MEF(50%)) increased from 125.3% to 130.2%, forced expiratory vital capacity 25% of the moment bit of gas flow (MEF(25%)) reduced from 86.9% to 71.7%; serum ceruloplasmin levels decreased from 690 mg/L to 180.6 mg/L; lung high-kV X-ray at 1st review showed that diffuse lung nodules had been absorbed and getting smaller than before treatment; chest CT showed that the distribution and number of small nodules at double lung fields decreased than before treatment. CONCLUSION: HGF combined with BMSC transplantation may have some potential role for the treatment of silicosis patients. PMID- 21619797 TI - [Expression of nuclear factor kappaB and tumor necrosis factor alpha in lung tissue of paraquat poisoned rats]. AB - OBJECTIVE: To investigate the expression of nuclear factor kappa B (NF-kgr;B) and tumor necrosis factor alpha (TNF-alpha) in lung tissue of acute paraquat poisoned rats. METHODS: 68 male Wistar rats were randomly divided into 2 groups: the control group (n = 8), the intoxication group (n = 60). On the 1st, the 3rd, the 7th, the 14th and the 28th day after intoxication, the expression of NF-kappaB p65 and TNF-alpha in lung tissue were detected by LSAB immunohistochemistry (IH) staining. Meanwhile, the level of malondialdehyde (MDA) in plasma, and lung homogenate, the content of malondialdehyde (HPY) in lung homogenate were detected. RESULTS: The levels of MDA in plasma on the 1st, the 3rd, the 7th day and in lung homogenate on the 1st, the 3rd day of the intoxication group [in plasma: (10.15 +/- 3.15), (6.97 +/- 1.65) and (5.44 +/- 0.66) nmol/ml; in lung homogenate: (10.20 +/- 2.43), (10.71 +/- 171) nmol/ml] were significantly higher than that of the control group [in plasma: (3.84 +/- 1.04) nmol/ml, in lung homogenate: (7.66 +/- 0.66) nmol/ml]. The content of HPY in lung homogenate on the 14th and the 28th day after intoxication [(19.98 +/- 2.86), (26.06 +/- 4.06) ug/0.1 g lung homogenate] were higher than that of the control group [(8.80 +/- 1.26) ug/0.1 g lung homogenate] significantly. The expression of NF-kappaB p65 and TNF-alpha in lung tissue were both significantly increased on the first day and the 3rd day of the intoxication group compared with the control group and weakened obviously after the 7th day. CONCLUSION: Acute paraquat poisoning can induce increased expression of both NF-kappaB p65 and TNF-alpha in lung tissue; the enhanced activity of NF-kappaB may take part in the process of pulmonary injury in PQ poisoning. PMID- 21619798 TI - [Effects of garlic oil, age and sex on n-hexane metabolism in rats]. AB - OBJECTIVE: To investigate effects of garlic oil (GO), age and sex on n-hexane metabolism in rats. METHODS: The Wistar rats were used as experimental animals. (1) Intragastric administration: n-hexane group (3000 mg/kg n-hexane), GO treated group (80 mg/kg GO ig. an hour earlier than 3000 mg/kg n-hexane), then blood was taken from tails of rats at 8, 12, 16, 20, 24, 28, 32 h points after n-hexane administration. (2) Intraperitoneal injection: n-hexane group (1000 mg/kg n hexane), GO treated group (80 mg/kg GO ig. an hour earlier than 1000 mg/kg n hexane), then took blood was taken from tails of rats at 8, 12, 16, 20, 24, 28 h points after n-hexane injection. (3) 7 rats each group of 6, 8, 10 weeks age were administrated by 3000 mg/kg n-hexane intragastrically, then were taken blood from tails at 16, 20, 24 h points after administration. (4) 7 male and 7 female rats of 8 weeks age were administrated by 3000 mg/kg n-hexane intragastrically, then were taken blood from tails at 16, 20, 24, 28 h points after administration. The gas chromatography was used to determine the metabolite 2, 5-hexanedione concentration of n-hexane in serum and 2, 5-hexanedione concentration was compared between GO and no GO treated rats, different ages and different sexes. RESULTS: (1) Intragastric administration: 2, 5-hexanedione concentrations in serum of n-hexane group and GO treated group had the peak 19.2 and 12.3 ug/ml at 20h and 24 h points. Compared with n-hexane group, the serum 2, 5-hexanedione concentration of GO treated group was lower at time points prior to peak and 2, 5 hexanedione eliminating process was slower after peak. (2) Intraperitoneal injection: effects of GO on the serum 2, 5-hexanedione concentrations was very similar to intragastric administration, 2, 5-hexanedione concentrations in serum of n-hexane group and GO treated group had the peak 15.0 and 6.7 ug/ml at 12 h and 16 h points. (3) Comparison of the serum 2, 5-hexanedione concentrations of different weeks age rats: The serum 2, 5-hexanedione concentrations of 6, 8, 10 weeks age rats were 25.5, 15.0, 12.8 ug/ml each (8, 10 weeks age significantly lower than 6 weeks age) at 16 h point; at 20 h point, they were 24.7, 18.3, 15.0 ug/ml each (10 weeks age significantly lower than 6 weeks age); at 24 h point, they were 11.0, 14.7, 8.1 ug/ml each (10 weeks age significantly lower than 8 weeks age). (4) Comparisons of the serum 2, 5-hexanedione concentrations of different sex rats: the serum 2, 5-hexanedione concentrations of male and female rats were 22.5, 17.2 ug/ml each at 16 h point (different significantly); at 20, 24, 28 h points, they were 27.6, 22.9 ug/ml, 24.6, 19.1 ug/ml, 19.1, 13.8 ug/ml each (different non-significantly). CONCLUSION: GO reduces production of 2, 5 hexanedione in serum generated by n-hexane in rats; the metabolic capacity of low age rats on n-hexane is stronger than high age ones. PMID- 21619799 TI - [Situation of pesticide poisoning in Huzhou from 2006 to 2009]. AB - OBJECTIVE: To understand the situations of pesticide poisoning in Huzhou and take preventive strategy and measures against the pesticide poisoning. METHODS: Case reports between 2006 and 2009 in the data base of reporting system for occupational diseases were computed by Excel for windows and statistical significance by SPSS12.0. RESULTS: A total of 2298 patients were reported from 2006 to 2009. Among them, the incidence of occupational poisoning accounted for 25.59% (588 cases), including 4 fatalities (fatality rate, 0.68%). Male patients (458 cases, 77.89%) were more than female ones (130 cases, 22.11%) in occupational pesticides poisoning. Summer and autumn were the most seasons in occupational pesticides poisoning occurring. The incidence of non-occupational pesticides poisoning accounted for 74.41% (1710, cases), including 112 fatalities (fatality rate, 6.55%). Female patients (952 cases, 55.67%) were more than male ones (758 cases, 44.33%) in non-occupational pesticides poisoning. 15 - 55 years were the highest incidences among non-occupational pesticides poisoning patients. Insecticides especially organophosphorus insecticides such as methamidophos, parathion, and omethoate comprised a higher proportion, accounting for 79.98% of the pesticides poisoning. CONCLUSION: The incidence and the fatality rate of occupational pesticide poisoning were reduced in the city. However, more attention should be paid to non-occupational pesticides poisoning. To decrease the numbers of pesticide poisoning and the risks of death, the relevant departments should take preventive strategy and measures against the pesticide poisoning. PMID- 21619800 TI - [Analysis on the incidence of coal workers' pneumoconiosis from 2003 to 2008 in a coal mining group]. AB - OBJECTIVE: Analyzed associations among the incidence of coal workers' pneumoconiosis from 2003 to 2008, jobs, exposure years and cumulative total dust exposure levels (CTE) and found the current characteristics of the mine incidence of pneumoconiosis disease. METHODS: collected the health care information of the new diagnosed pneumoconiosis of underground mine workers from 2003 to 2008 and the dust monitoring data of underground mine from 1949 and estimated the personnel cumulative total dust exposure levels (CTE); analyzed the incidence features of the new diagnosed pneumoconiosis. RESULTS: The rates of health surveillance of workers were gradually improved from 2003 to 2008 and 296 new coal workers pneumoconiosis were diagnosed. The total incidence was 0.57%, and the average annual rate was 0.32%. Among the new diagnosed cases, phase I accounted for 90.5% and the 87.2% from coal mine drillers. The shortest exposure period was 3 years and the longest was 38 years, and the cumulative total dose of dust was varied between 86.1 and 4926 mg/m(3) per year. The total dust accumulated limited dose was calculated by the percentile method to prevent 99% of miners from pneumoconiosis, which was 120.6 mg/m(3) per year, so we suggested that the exposure years should be shorter than 13 years under the current working conditions. CONCLUSIONS: Preventive coal workers' pneumoconiosis should be focused on mine drillers and their limited exposure years should be within 13 years. PMID- 21619801 TI - [Investigation about prevention behavior for dust workers in machinery, ceramic, and metallurgy industry]. AB - OBJECTIVE: The purposes of this thesis were to study the behavior about workers exposed to dust and provide scientific basis for health promotion. METHODS: We designed a questionnaire and carry it on the 746 dust workers in the 3 representative corporations of Machinery, Ceramic, and Metallurgy Industry. All data were input into computer. And a database was established with Excel. SPSS11.5 statistical analysis software was used to analyze the influence on protecting behavioral between the application of qualifications, different jobs, training or protection, and other aspects etc. RESULTS: The rates were 94.4% and 75.3% about the regular physical examination and requirements for protective equipment. The rate of choosing an effective way of protection was generally low (15.4%). There was significant difference for among different educational background workers (P < 0.01). The rates of choosing an effective way of protection (20.3%), the regular physical examination (98.3%) and requirements for protective equipment (86.4%) in the dust workers who participated in the training of dust protection were superior than those who did not participated in the training. There was the significant difference (P < 0.05, P < 0.01). There was the significant difference for the rate of effective way of protection, regular physical examination, and requirements for protective equipment among the different corporations (P < 0.05). CONCLUSIONS: Dust workers' using rate about the choosing an effective way of protection was generally low in Machinery, Ceramic, and Metallurgy Industry. Those who were not educated had a lower using rate about the protection behavior, regular physical examination, and requirements for protective equipment than those educated. PMID- 21619802 TI - [MRI image analysis of 1,2-dichloroethane chronic toxic encephalopath]. AB - OBJECTIVE: To study the MRI features of 1,2-dichloroethane Chronic Toxic Encephalopathy of 10 cases. METHOD: 10 cases were examined by MRI, slice thickness 8 mm, layer from 2 mm, axial and coronal line scan, T1WI, T2WI, FLAIR imaging. RESULTS: 10 cases show varying degrees of abnormal signal of white matters, low signal intensity on T1WI, high signal intensity on T2WI and FLAIR. MRI could also show extensive abnormal signal in cerebral white matter although the toxic manifestation is mild to moderate. Therefore the symptoms and the shows of MRI could be inconsistent. CONCLUSION: Combined with a history of exposure, the show of varying degrees of abnormal signal of white matter in 1,2 dichloroethane Chronic Toxic Encephalopathy cases are characteristic. PMID- 21619803 TI - [Analysis on bone marrow feature of 56 clinic cases of benzene poisoning]. AB - OBJECTIVE: To explore the bone marrow feature of hemopoietic system injured by benzene through analyzing 56 benzolism cases. METHODS: The 56 benzolism cases were divided into mild poisoning group, midrange poisoning group, aplastic anemia group, pancytopenia group and leukemia group. All cases progressed bone marrow aspiration and smear, and counted hundred karyocytes by Wright-Giemsa tinct bone marrow smear to classification and observe the cells' feature. RESULTS: The megakaryocytes and the extent of bone marrow hyperplasia were decreased by turns of mild poisoning group, midrange poisoning group and aplastic anemia group. The archaeocytes and juvenile cells proliferation in mild poisoning group and midrange poisoning group were inhibited and occurred cell paramorphia which related to intoxication. Comparing with the other groups and normal reference value, the pancytopenia group's percentage of bone marrow cells in karyocytes was significantly decreased (P < 0.01, P < 0.05) and the leukemia group's percentage of bone marrow cells in karyocytes was significantly increased (P < 0.01). The proportion of cell paramorphia and nucleus malformation of granulocytes and red blood cells in pancytopenia group and leukemia group were increased, especially in leukemia group. CONCLUSION: We saw the inhibition of archaeocytes and juvenile cells proliferation and some cell paramorphia appearances in mild poisoning and midrange poisoning cases of chronic benzolism. The abnormality changes which can be seen in bone marrow of severe benzolism cases were corresponding with the clinical classification. PMID- 21619804 TI - [Management of phenol burn cases combined with poisoning]. AB - OBJECTIVE: To explore the therapeutic processing and nursing for phenol burn patients combined with poisoning. METHODS: The clinical data of 42 phenol burn cases admitted in our hospital from 1998 to 2010 were reviewed retrospectively. The general clinical manifestations, treatment and nursing methods, and the treatment results of these cases were analysed. RESULTS: Out of 42 phenol burns patients, the total burn surface area (TBSA) of 22 cases were <= 5%, 10 cases' TBSA were 6% ~ 10%, and 5 cases' > 20%. 36 patients showed systemic poisoning symptoms 1 ~ 2 hours after injury, the severity and enduring period of which correlated with TBSA. Patients whose TBSA > 10% appeared obviously dysfunction of the liver and kidney. The therapeutic methods included wound management, compensation of fluid, diuresis, alkalized urine, large dose of vitamin C and glucocorticoid, early surgery, blood purification, etc. 39 of 42 cases were successful cured, while the other 3 cases died. CONCLUSIONS: Phenol burn often complicates systemic poisoning. In the treatment of patients with this special chemical burn, it is key scheme to block phenol absorption and promote elimination of already absorbed poison as early as possible. The comprehensive management and nursing should be necessary for supporting functions of important organs. PMID- 21619805 TI - [High-resolution melting: a analysis for genotyping of MDR1 C3435T in benzene exposed workers]. AB - OBJECTIVE: Using high resolution melting (HRM) to analysis MDR1 C3435T in people exposed to benzene. METHODS: Restriction fragment length polymorphism (RFLP) was utilized to detect the polymorphism of MDR1 3435 in 121 benzene-exposed workers, and the results were compared with the HRM in 10% samples and were confirmed with direct sequencing for six people in them. RESULTS: By direct sequencing, consistent results of benzene-exposed workers with RFLP or HRM were got. The new high resolution melting curve analysis is more efficient, more convenient, and cheaper than RFLP. CONCLUSION: High-resolution melting analysis provides a valid approach to efficiently detect DNA genetic diagnosis, which is suitable for detect susceptible genes in occupational surveillance. PMID- 21619806 TI - [Recent research on occupational paraquat poisoning]. PMID- 21619808 TI - [Targeting ergonomics interventions towards promoting occupational health and safety]. PMID- 21619807 TI - [Acute injury of nano titanium dioxide particles on lung]. PMID- 21619809 TI - [The analysis of factors influencing the work related musculoskeletal disorders of oil drillers]. AB - OBJECTIVE: To investigate the morbidity and influence factors of work-related musculoskeletal disorders (WMSD) in oil drillers. METHODS: The modified Nordic Standardized Questionnaire for WMSD was used to perform the epidemiological investigation in 860 oil drillers. RESULTS: The predominant sites of WMSD in the oil drillers were waist (50.6%), neck (29.8%) and shoulder (23.4%), respectively. There were significant differences of the WMSD morbidities in all body sites except for wrist and hip among groups with different working years (P < 0.05 of P < 0.01). The WMSD morbidities in the neck, shoulder, back and waist of oil drillers working for more than 40h a week were significantly higher than those of oil drillers working for less than 40 ha week (P < 0.05). The results of logistic regression analysis revealed that some occupational factors, i.e. keeping stoop and stand for long time, repeating trunk bend, keeping awkward lift posture and lifting the heave objects, were the risk factors for WMSD at waist (P < 0.05), also some management factors, i.e. overtime work and inadequate rest, were the risk factors for WMSD at waist (P < 0.05). CONCLUSION: The WMSD appears to be a serious ergonomic problem in oil drillers, it is necessary to correct working posture, reduce working load, improve organizational management, and encourage a workplace health program with regular work and rest. PMID- 21619810 TI - [Experimental simulated study on the break for manual lifting task by surface electromyography and electrocardiogram]. AB - OBJECTIVE: To investigate the recovery duration of manual lifting tasks. METHODS: 30 college student volunteers with gender equally distributed were involved in the simulated stoop-, squart- and semi squart-lifting tasks in the laboratory. Signals of electromyography (EMG) at the back muscles (left and right thorax m. erector spinae, LT(10) and RT(10); left and right lumbar m. erector spinae, LL(3) and RL(3)) and electrocardiography (ECG) were recorded during the rest intervals. The percentage of maximum voluntary electrical activation (MVE%), EMG median frequency (MF), and heart rate (HR) were analyzed to predict the recovery time required. RESULTS: During the break, MVE% at 4 back muscles of all the male subjects were recovered to resting level within 5 minutes and 91.8% were within 2 minutes. Except for RL(3) after squat- and stoop-lifting task, the MVE% of female subjects went back to resting level within 3 minutes and 67.8% were within 2 minutes. The MF of male subjects were recovered in 10 minutes and 74.9% within 2 minutes, with the exception of RL(3) after stoop- and squat-lifting tasks. All the female MF were recovered in 8 minutes and 83.4% were within 2 minutes. However, during the last 20 minute break after the lifting tasks, the MVE% and MF of all male and female subjects were recovered, while HR came back to resting level within 14 and 5 minutes, respectively. CONCLUSIONS: For the three manual lifting tasks with 25% MVC workload lasting for 4.5 min, a break of 15 minutes is sufficient for most erectors spinae to recover from local muscle load. However, in terms of the HR, a break of 15 minutes is enough for the recovery of systematic load. PMID- 21619811 TI - [A field study on the work load and muscle fatigue at neck-shoulder in female sewing machine operators by using surface electromyography]. AB - OBJECTIVE: To study neck and shoulder work-related muscle fatigue of female sewing machine operators. METHODS: 18 health female sewing machine operators without musculoskeletal disorders work in Beijing garment industry factory as volunteers in participate of this study. The maximal voluntary contraction (MVC) and 20% MVC of bilateral upper trapezium and cervical erectors spinae was tested before sewing operations, then the whole 20 time windows (1 time window = 10 min) sewing machine operations was monitored and the surface electromyography (sEMG) signals simultaneously was recorded after monitoring the 20%MVC was tested. Use amplitude analysis method to reduction recorded EMG signals. RESULTS: During work, the median load for the left cervical erector spinae (LCES), right cervical erector spinae (RCES), left upper trapezium (LUT) and right upper trapezium (RUT) respectively was 6.78 +/- 1.05, 6.94 +/- 1.12, 5.68 +/- 2.56 and 6.47 +/- 3.22, work load of right is higher than the left; static load analysis indicated the value of RMS(20%MVC) before work was higher than that value after work, the increase of right CES and UT RMS(20%MVC) was more; the largest 20%MVE of bilateral CES occurred at 20th time window, and that of bilateral UT happened at 16th. CONCLUSIONS: The work load of female sewing machine operators is sustained "static" load, and work load of right neck-shoulder is higher than left, right neck-shoulder muscle is more fatigable and much serious once fatigued. PMID- 21619812 TI - [An accident of trichlorosilane explosion]. PMID- 21619813 TI - [The interactive effect of job task and psychosocial factors on work-related musculoskeletal disorders]. AB - OBJECTIVE: To investigate the interactive effect of job task and psychosocial factors on the outcomes of musculoskeletal disorders. METHODS: 653 workers from different type of manufacturing industries and administration office recruited in a cross-sectional epidemiological survey. The Quick Exposure Check (QEC) was applied to assess the ergonomic load of job task, Job Content Questionnaire (JCQ) for identifying psychological characteristics, and Nordic Standardized Questionnaire for investigating outcomes of WMSDs. RESULTS: The prevalence of WMSD in shoulder, upper back, lower back and hand/wrist were significantly different under a variety of combined job task and psychosocial characteristics (P < 0.05 or P < 0.01). The more physical and psychological loads, the higher prevalence of WMSDs were revealed. By using multivariate analyses, a potential interactive effect was found in terms of the WMSDs symptoms in hand/wrist, shoulder, upper back and lower back after adjusted by work year, age, and gender. CONCLUSIONS: Higher physical load and greater psychosocial risk are more frequent self-reported symptoms of WMSDs than those of lower exposures. Ergonomic intervention strategies aimed at reducing the incidence of WMSDs should not only be focused on control of physical work factors but also psychosocial risks of relevance. PMID- 21619814 TI - [Survey on prevalence of occupational musculoskeletal disorders among workers in a shipyard]. AB - OBJECTIVE: To study the prevalence of occupational musculoskeletal disorders (OMSDs) in a shipyard and explore the relationship between type of work, age, length of service and OMSDs by cross sectional epidemiological survey. METHODS: 1570 workers from a shipyard were selected as survey objects, and a revised Northern Europe Standardized Questionnaire was used to investigate OMSDs. 253 technicians and clerks in the shipyard were selected as controls. RESULTS: OMSDs in these workers primarily located in waist, neck and shoulder, prevalence were 58.0%, 54.6% and 44.3% respectively; There were significant differences among workers of different types of work (P < 0.05) in prevalence and the cumulative parts number of OMSDs. The control group in this study also showed a higher prevalence of neck, shoulder and waist OMSDs. Female workers had higher prevalence of OMSDs in neck and shoulder compared with male workers (P < 0.05). As well there was significant correlation between age and OMSDs prevalence except the wrist and ankle disorders (P < 0.05), OMSDs prevalence increased with years. In addition, there was correlation between length of service and OMSDs prevalence, the group of <= 5 years had the lowest prevalence and the prevalence of low-back, neck and shoulder increased with service years. CONCLUSION: The OMSDs in workers engaged in shipyard are serious, the length of service, age, gender and type of work seem to be the risk factors, and it seems reasonable that the ergonomics intervention in the above aspects should be necessary for the prevention of OMSDs. PMID- 21619815 TI - [Analysis of musculoskeletal disorders, work load and working postures among manufacturing workers]. AB - OBJECTIVE: To analyze the distribution of the musculoskeletal disorders, work load and working postures in different factories, gender, education levels, age and working years among manufacturing workers. METHODS: In a cross-sectional study of 5134 manufacturing workers in 12 factories, the morbidities for musculoskeletal disorders in one year period were measured with questionnaires. RESULTS: The morbidities for musculoskeletal disorders in body sites: waist, neck, shoulder, wrist, ankle/feet, knee, hip/buttocks and elbows were 59.7%, 47.9%, 38.1%, 33.7%, 26.9%, 25.4%, 15.2%, and 14.9%, respectively in one year period. There were significant differences of morbidities for musculoskeletal symptoms in body sites of workers among different factories (P < 0.05 or P < 0.01). The morbidities of musculoskeletal symptoms in elbows, waist, wrists and ankle/feet of the workers in refractory material and chemical fiber factories were higher than those in other factories, the morbidities for musculoskeletal symptoms of workers in garments and diamond factories were lower than those in other factories. The morbidities for musculoskeletal symptoms in neck, shoulders and wrists of female workers were significantly higher than those of male workers (P < 0.01). There were significant differences of the morbidities for musculoskeletal symptoms in body sites among workers with different educational levels (P < 0.05 or P < 0.01). There were significant differences of the morbidities for musculoskeletal symptoms in neck, shoulders, wrists, hip/buttocks and knee among groups with different age or different working years (P < 0.01), and the morbidities for musculoskeletal symptoms increased with age and working years. The proportions of unhealthy working postures and high working load among workers in refractory material and chemical fiber factories were higher; but those in garments and diamond factories were lower. CONCLUSION: The morbidities for musculoskeletal symptoms in waist, neck, shoulder and wrists of workers in manufacturing workers were higher; the gender, education level, age and working years could influenced the morbidities for musculoskeletal disorders. PMID- 21619816 TI - [Musculoskeletal disorders and risk factors of workers in a coal mine]. AB - OBJECTIVE: To explore the prevalence characteristics and influence factors related to occupation and individuals for musculoskeletal disorders of workers in Chinese mines. METHODS: In a cross-sectional study of 1900 coal miners from a coal mine, the Standardized Nordic Questionnaire was used to assess the musculoskeletal disorders, and logistic regression analysis was performed to analyze the correlation between the occupational factor and he musculoskeletal disorders. RESULTS: During the past year, 1205 miners of 1537 miners (78.4%) complained of the musculoskeletal disorders. The morbidity of lumbago was 59.5%. The morbidity of the musculoskeletal disorders in different body sites of the miners increased significantly with age (P < 0.05). The morbidity of the musculoskeletal disorders in the underground workers was significantly higher than that in the ground workers. According to logistic regression analysis, the repetitive operation and awkward posture were the risk factors for the musculoskeletal disorders in neck, shoulder and upper limbs; the repetitive operation, moving heavy substance and stooping posture were related significantly to lumbago; the musculoskeletal disorders in lower limbs were associated with the long standing and awkward posture. CONCLUSION: There is significant correlation between the occupational factors and the musculoskeletal disorders for coal miners. PMID- 21619817 TI - [The effect of DNA methyltransferase 1 low expression on the global genome DNA methylation status of 16HBE cell]. AB - OBJECTIVE: To construct DNA methyltransferase 1 (DNMT1) low expression 16HBE cell line and observe the variation of cell cycle and global genomic DNA methylation. METHODS: The method of Lenti-virus induced RNA interference was applied to introduce four different shRNA fragment into 16HBE cells. Flow cytometry and 5-mC immunofluorescence methods were used to observe the cell cycle and global DNA methylation status of DNMT1 low expression 16HBE cells. RESULTS: The DNMT1 protein relative expression level of 16HBE-shDNMT1-4 cell line was down regulated about 44% (P < 0.05) compared with the control. No obvious differences of cell cycle and global genome DNA methylation status were observed between the 16HBE and 16HBE-shDNMT1. CONCLUSION: The DNMT1 gene low expression cell is successfully constructed, and there are no obvious changes happened on the cell cycle and global genomic DNA methylation. PMID- 21619818 TI - [The expression changes of N-methyl-D-aspartic acid receptor in hippocampus of offspring from female rats exposed to aluminum in the pregnancy and lactation]. AB - OBJECTIVE: To investigate the effects of aluminum on learning and memory and the expression of N-methyl-D-aspartic acid receptor (NMDAR) of hippocampus in offspring from female rats exposed to Al in the pregnancy or lactation, and to explore the mechanism of toxic effects of Al on central nervous system (CNS) during development. METHODS: The pregnant Wistar rats were randomly divided into 3 groups based on their body weight, i.e. control group was exposed to distilled water, low exposure group (0.2%AlCl3) and high exposure group (0.4%AlCl3) were exposed orally to AlCl3 in pregnancy and lactation for 6 weeks, 10 rats each group. Aluminum content in blood and brains was determined by atomic absorption spectrophotometry (AAS). Platform experiment was used to detect the abilities of learning and memory. The expression levels of NMDARs were detected by western blot assay. RESULTS: The Al content in blood and brains of rats in exposure groups increased significantly with Al dose, as compared with the control group (P < 0.05). In platform experiment, the incubation periods of rats in low and high exposure groups were (202.71 +/- 81.99) and (19.67 +/- 8.44) s respectively, which were significantly lower than that [(300.00 +/- 0.00) s] in control group (P < 0.01), but the mistake times of rats in low and high exposure groups were 1.43 +/- 0.85 and 2.47 +/- 0.99 respectively, which were significantly higher than that (0.00 +/- 0.00) in control group (P < 0.01). The Al exposure could change the proportion of NMDAR subtypes, the expression levels of NR1 and NR2B in hippocampus of newborn rats in low and high exposure groups were 25.22 +/- 0.68, 81.23 +/- 15.37 and 24.75 +/- 0.71, 56.63 +/- 7.82, respectively, which were significantly lower than those (31.69 +/- 3.44, 107.61 +/- 9.05) in control group (P < 0.05). CONCLUSION: Aluminum exposure in pregnancy and lactation could reduce the abilities of learning and memory in newborn rats, and change the proportion of NMDAR subtypes. The reduced NR1 and NR2B expression levels may be one of important mechanisms to influence the abilities of learning and memory in offspring. PMID- 21619819 TI - [Evaluation of different oxygen therapies on therapeutic effects in rats with acute carbon dioxide poisoning]. AB - OBJECTIVE: To study therapeutic effects by using different oxygen therapies in rats with acute carbon dioxide poisoning, to select the best oxygen therapy technology for patients with acute carbon dioxide poisoning on the spot. METHODS: Sixty healthy male Sprague-Dawley rats were randomized into normal control group, carbon dioxide exposure group, hyperbaric oxygen treatment group (pressure 2 ATA, FiO(2)100%), high concentration of atmospheric oxygen treatment group (FiO(2)50%), low concentration of atmospheric oxygen treatment group (FiO(2)33%). After treated with different oxygen in rats with acute carbon dioxide poisoning, arterial pH, PO2 and PCO2 of rats were detected, in addition observe pathological changes of lung tissue and brain tissue. RESULTS: The arterial pH (7.31 +/- 0.06) and PO2 [(68.50 +/- 15.02) mm Hg] of carbon dioxide exposure group were lower than those of control group [pH (7.42 +/- 0.02) and PO2 (92.83 +/- 8.27) mm Hg], PCO2 [(71.66 +/- 12.10) mm Hg] was higher than that of control group [(48.25 +/- 2.59) mm Hg] (P < 0.05); the arterial pH (hyperbaric oxygen treatment group 7.37 +/- 0.02, high concentration of atmospheric oxygen treatment group 7.39 +/- 0.03, low concentration of atmospheric oxygen treatment group 7.38 +/- 0.02) and PO2 of oxygen treatment groups [hyperbaric oxygen treatment group, high concentration of atmospheric oxygen treatment group, low concentration of atmospheric oxygen treatment group were (82.25 +/- 12.98), (84.75 +/- 11.24), (83.75 +/- 16.77) mm Hg, respectively] were higher than that of carbon dioxide exposure group, PCO2 [hyperbaric oxygen treatment group, high concentration of atmospheric oxygen treatment group, low concentration of atmospheric oxygen treatment group were (52.25 +/- 4.95), (51.75 +/- 4.82), (52.66 +/- 5.61) mm Hg, respectively] was lower than that of carbon dioxide exposure group (P < 0.05); there was no significant difference of the arterial pH, PO2 and PCO2 between oxygen treatment groups and control group (P > 0.05); there was no significant difference of the arterial pH, PO2 and PCO2 among oxygen treatment groups (P > 0.05). There was large area of bleeding of lungs in rats with carbon dioxide poisoning, the bleeding of lungs in rats with high concentration of atmospheric oxygen treatment and low concentration of atmospheric oxygen treatment was better than the rats with carbon dioxide poisoning, there was no abnormal appearance of lungs in rats with hyperbaric oxygen treatment. The light microscope observation showed that there were diffuse bleeding and exudation of lungs in rats with carbon dioxide poisoning, the bleeding and exudation of lungs in rats with high concentration of atmospheric oxygen treatment and low concentration of atmospheric oxygen treatment were better than the rats with carbon dioxide poisoning, there were only minor bleeding and exudation of lungs in rats with hyperbaric oxygen treatment. There was no difference of brain in anatomy and microscopy among all groups, there were no significant bleeding, edema, cell degeneration and necrosis. CONCLUSIONS: Lung pathology in acute carbon dioxide poisoning rats with hyperbaric oxygen treatment is better than the rats with high concentration of atmospheric oxygen treatment and low concentration of atmospheric oxygen treatment, there is no significant difference of effect between high concentration of atmospheric oxygen treatment group and low concentration of atmospheric oxygen treatment group, however, the results of blood gas analysis and lung pathology than the exposure group improved, so qualified medical unit for hyperbaric oxygen therapy as soon as possible, hyperbaric oxygen treatment facilities in the absence of circumstances, the emergency treatment of early oxygen is also a good measure. PMID- 21619820 TI - [Relationship between MnSOD polymorphisms and susceptibility of chronic poisoning exposed to manganism occupationally]. AB - OBJECTIVE: To study the relationship between polymorphisms of MnSOD and the susceptibility of chronic poisoning exposed to manganism occupationally. METHODS: In a study of case-control, genotypes were determined by PCR-RFLP in 164 patients with chronic occupational mangamism poisoning and 328 controls with age- and sex matched for MnSOD 9Ala-Val. RESULTS: There was a significant difference in the frequency of MnSOD 9Ala-Val at V locus mutant allele between cases and controls (chi(2) = 15.225, P < 0.01, 95%CI = 1.43 ~ 3.00). Individuals with the genotype VV had a 1.30 of risk increase of occupational chronic manganism poisoning compared with the the genotype AV or AA (OR = 2.30, 95%CI = 1.52 ~ 3.49, P < 0.05). CONCLUSION: The MnSOD polymorphisms may be related with the susceptibility to chronic occupational manganism poisoning, the risk of chronic occupational manganism poisoning increases in carriers with genotype VV at MnSOD 9Ala-Val locus. PMID- 21619821 TI - [The effects of infrasound on the blood coagulation function of guinea pigs]. AB - OBJECTIVE: To study the change of the blood coagulation function of guinea pigs exposed to 16 Hz/120 dB, 16 Hz/125 dB infrasound and to explore the mechanism of circulation system damage. METHODS: Seventy-two guinea pigs were divided into 3 groups: the control group, the group exposed to 16 Hz/120 dB infrasound for 1.5 h a day and the group exposed to 16 Hz/125 dB infrasound for 1.5 h a day. Each exposure group was divided into 4 sub-groups (8 guinea pigs a sub-group) which were exposed to infrasound for 1, 7, 14 and 21 d, respectively. The coagulation function and serum nitric oxide (NO) were measured for control group and all sub groups after exposure to infrasound. RESULTS: The prothrombin time (PT), international normalized ratio (INR) and serum NO of group exposed to 16 Hz/125 dB infrasound were (31.16 +/- 3.05) s, 2.53 +/- 1.21 and (88.304 +/- 52.601) umol/L, respectively, which were significantly higher than those [(21.36 +/- 0.10) s, 1.65 +/- 0.07 and (30.943 +/- 26.864) umol/L] of control group (P < 0.05). PT and INR of sub-groups exposed to 16 Hz/125 dB infrasound for 14 and 21 d were significantly higher than those of control group. NO of sub-groups exposed to 16 Hz/125 dB infrasound for 1 week and 2 weeks were significantly higher than that of control group (P < 0.05), but NO of sub-group exposed to 16 Hz/125 dB infrasound for 3 weeks decreased slightly. CONCLUSION: The blood coagulation function of guinea pigs exposed to 16 Hz/125 dB infrasound decreased, PT and INR may be used as the indexes to assess of blood coagulation function change induced by the infrasound exposure. PMID- 21619822 TI - [Study on the prognosis of patients with acute paraquat intoxication]. AB - OBJECTIVE: To study the correlation factors of acute paraquat intoxication prognosis. METHODS: The early paraquat concentration in plasma and urine, leukocyte count, hepatic and renal function, amylase, electrolyte and the parameters of arterial blood gas were analyzed retrospectively in 111 patients with acute paraquat intoxication. RESULTS: 43 cases (38.7%) of all the 111 patients survived and the other 68 cases (61.3%) died. The patient, whose paraquat concentration was not more than 8.0 ug/ml in plasma and 276.0 ug/ml in urine, could survive. But some patients could die, only if there was no paraquat found in plasma. The paraquat levels in plasma and urine were significantly lower in survivors [(0.82 +/- 1.70), (28.12 +/- 51.17) ug/ml] than in nonsurvivors [(9.32 +/- 12.04), (384.53 +/- 597.93) ug/ml, respectively] (P < 0.01). The levels of leukocyte count, serum creatinine, aspartate aminotransferase (AST), and amylase were significantly higher in nonsurvivors than in survivors (P < 0.05, P < 0.01). In addition, metabolic acidosis was easier to appear in nonsurvivors. The multiple logistic regression analysis indicated that the paraquat concentration in plasma and urine, leukocyte count, creatinine and base excess were all related to survival. CONCLUSION: The higher paraquat concentration in plasma and urine, leucocytosis, renal dysfunction and metabolic acidosis are all important factors for the prognosis of paraquat intoxication. PMID- 21619823 TI - [A cross-sectional study on the industrial noise over-limit status in Guangzhou factories]. AB - OBJECTIVE: To investigate the industrial noise over-limit status of the worksites in Guangzhou factories, so as to promote the prevention and control of occupational noise hazards. METHODS: 211 factories in Guangzhou were monitored and investigated. The analysis and assessment were developed for the properties and size of the factories, the districts of the factories being located, the industries of the factories being classified and the date of monitoring. RESULTS: In this understudied factories, most of them were national-owned and joint ventures, medium size, located in urban, and mainly involved in the industries of manufacturing of motor vehicle, shipping, electron and electric equipment, and the industries of petroleum and chemicals. The prevalence of noise over-limit was higher in joint-ventures (36.0%) and private-run enterprises (31.2%). The over limit status mainly presented in industries of textile, food and beverage processing, and leather producing, with getting prevalence of over-limit 46.7%, 43.1% and 41.3% respectively. Subsequence were industries of manufacturing of electron and electric equipment, motor vehicle and shipping, and industries of printing and goods producing for culture and sports, with the prevalence for all > 35%. Factories monitored during spring and summer also had higher prevalence of noise over-limit. The similar results were got after adjustment for each other using multivariable regression. The most common over-limit sites mainly focused on the operation of cutting and sawing, milling and planing, pressing, riveting, drilling, jointing, assembling and quality inspecting in industries of mechanism processing and manufacturing, on quality inspecting and packing in industries of pharmacy and food and beverage manufacture, on spinning and scutching in textile industry, and on cleaning and maintaining as assistant jobs, and patrolling and inspecting air-press machine, ventilation machine, dynamotor and pump. CONCLUSION: Noise in Guangzhou factories widely exists with different industries and districts. To strengthen noise occupational hazards prevention and control for the high risk districts, industries and worksites should be the key job in the future. PMID- 21619824 TI - [Effects of intervene on occupational health knowledge, attitude and practice of people with occupational disease risk in private enterprises]. AB - OBJECTIVE: To evaluate the effects of health promotion on occupational health based on the changes in knowledge, attitude and practice (KAP) before and after intervention of people with occupational disease risk in private enterprises. METHODS: 202 people with occupational disease risk in private leather enterprises of Wenzhou were surveyed, who were rechecked with the same questionnaire after three months intervention. RESULTS: The knowledge, attitude and practice scores (9.34 +/- 2.57, 7.79 +/- 2.58 and 7.24 +/- 2.50, respectively) of post intervention group were significantly increased more than those of pre intervention (8.06 +/- 2.71, 7.63 +/- 2.67, 7.11 +/- 2.60, respectively) (P < 0.01, P < 0.05). The net increases of knowledge, attitude and practice scores were significantly different with different length of service, educational level, registered residence and training experience (P < 0.05, P < 0.01). CONCLUSIONS: Health promotion could increase knowledge, attitude and practice levels; The effect of intervention on people with short length of service, low educational level, coming from country and had not attended training is significant. PMID- 21619825 TI - [Abnormal increase in CD8(low) T lymphocyte in patients with occupational chronic lead poisoning]. AB - OBJECTIVE: To analyze the changes in CD8(low) T lymphocyte subsets in patients with occupational chronic lead poisoning. METHODS: Flow cytometric analysis was used to count the numbers of CD8+ cells. 23 patients with occupational chronic lead poisoning and 20 controls were examined. RESULTS: Compared with control group (8.21% +/- 3.02%), the CD8(low) T lymphocyte (12.98% +/- 5.62%) were significantly increased in patients with occupational chronic lead poisoning. CONCLUSION: Although the ratio of CD+ T lymphocyte is normal, the CD8 level is significantly decreased. The increase of CD8(low) T lymphocyte may be an important phenomenon of immuno-injury induced by lead. CD8(low) T lymphocyte could be an new direction for research of lead immuno-toxicity. PMID- 21619826 TI - [Effect on expression of vascular smooth muscle cell calcium channel in rats exposed in low temperature]. AB - OBJECTIVE: To observe the expression of vascular smooth muscle cells calcium channel alpha1C subunit (LTCCalpha1C) in rats exposed in low temperature. METHODS: Cold-induced hypertension was established and blood pressure was measured every two weeks. The mRNA expression of L type calcium channel alpha1C was determined by RT-PCR. RESULTS: The blood pressure of the rats exposed to cold environment increased. The blood pressure of experimental groups [(102.8 +/- 2.25) mm Hg] began to increase from the first two weeks, compared with the control group [(89.2 +/- 3.73) mm Hg], there were significant difference (P < 0.05). The blood pressure of experimental groups were (114.5 +/- 4.21), (121.9 +/ 3.03) mm Hg respectively at 4, 6 weeks. Compared with the control group, the expression of LTCCalpha1C mRNA of the cold exposure group increased significantly (P < 0.01). There was a significant correlation between the expression of LTCCalpha1C mRNA and the blood pressure of the rats (r = 0.86, P < 0.01). CONCLUSION: Repeated cold exposure can establish cold-induced hypertension, and the level of vascular smooth muscle cells LTCCalpha1C expression increase. PMID- 21619827 TI - [Clinical study of dynamic change of inflammatory factors in serum of acute paraquat rats]. AB - OBJECTIVE: To investigate the change of inflammatory factor in lung tissue of acute paraquat (PQ) poisoned rats. METHODS: hundred SD rats were randomly divided into two groups: the normal control group (NC group, n = 10) and the PQ group (n = 80). The 1 ml saline was administered once in normal control group. The PQ group was administered with 25 mg/kg 1% PQ by intraperitoneal injection to establish the model of PQ induced renal injury. At six hours, at the first, the third and the seventh day the PQ group were sacrificed, while at the first day the normal control group was sacrificed. The level of normal tumor necrosis factor-alpha (TNF-alpha), interleukin-2 (IL-2), IL-6 in serum of rats were detected. Meanwhile, pathological changes of the renal were examined under optical microscope. RESULTS: Histopathological findings of an earlier, a large number of patients edema clearly inflammatory cell infiltration. Compared with the control group, PQ exposure of serum TNF-alpha, IL-2, IL-6, the level at each time point were elevated. PQ treated group 6 h and 1, 3, 7 d when the IL-2 levels were (2.16 +/- 0.65), (2.95 +/- 1.02), (3.05 +/- 1.12), (2.21 +/- 0.62) ug/L, IL 6 were (62.5 +/- 8.6), (85.6 +/- 13.5), (90.3 +/- 15.6), (65.3 +/- 9.1) ng/ml, TNF-alpha were (1.95 +/- 0.53), (2.86 +/- 0.92), (3.15 +/- 1.02), (2.06 +/- 0.71) ug/L, compared with the control group, are significantly higher, the differences were statistically significant (P < 0.01). CONCLUSION: acute PQ poisoning serum TNF-alpha, IL-2, IL-6 levels were significantly increased both early and late inflammatory factors involved in PQ poisoning the pathogenesis of renal injury. PMID- 21619828 TI - [Clinical study on simvastatin treatment of pulmonary hypertension in patients with coal worker's pneumoconiosis]. AB - OBJECTIVE: To observe simvastatin treatment of pulmonary hypertension in patients with coal workers pneumoconiosis (CWP). METHODS: 96 CWP patients with pulmonary hypertension were randomly divided into treatment and control groups. The control group was treated with 2.5 mg warfarin, once a day for four months; the treatment group was treated with 20 mg simvastatin, taken in evening, for 4 months. 6 min walking distance (6MWD) test and inspection pulmonary artery pressure were measured by echocardiography before and after treatment. RESULTS: In the treatment group, the 6MWD were (258 +/- 26) m after treatment and (225 +/- 19) m before treatment, respectively. Compared with control group, pulmonary artery pressure was (41 +/- 9) mm Hg in the treatment group before treatment, (36 +/- 3) mm Hg in the treatment group after treatment, and (39 +/- 5) mm Hg in control group, respectively, the difference was statistically significant (P < 0.05). CONCLUSIONS: Simvastatin can improve pulmonary hypertension in coal workers pneumoconiosis, and shows a definite curative effect. PMID- 21619829 TI - [A review on the reproductive health of males exposed to radiation by shipping radar microwave]. PMID- 21619830 TI - [A meta analysis on neuro-behavioral function of the workers exposed to manganism occupationally]. PMID- 21619831 TI - [Six cases injured by high pressure print spraying]. PMID- 21619832 TI - [Investigation on the occupational hazard of noise in a hydroelectric plant]. PMID- 21619833 TI - [The effects of ammonium perchlorate on thyroid function and mRNA expression of thyroglobulin and thyroperoxidase]. AB - OBJECTIVE: To investigate the effects of ammonium perchlorate (AP) on thyroid functions and mRNA expression levels of thyroglobulin (Tg) and thyroperoxidase (TPO) genes of rats. METHODS: Thirty SD male rats were randomly divided into six groups: control group, iodine-deficient group, low dose AP group (130 mg/kg), moderate dose AP group (260 mg/kg), high dose AP group (520 mg/kg) and high iodine-combined group. After the rats were exposed orally for 90 days, serum free thyroxine (FT(4)), free-triiodothyronine (FT(3)) and thyroid stimulating hormone (TSH) were measured using radioimmunoassays. mRNA expression levels of thyroglobulin (Tg) and thyroperoxidase (TPO) genes were detected by real-time quantitative PCR. RESULTS: Serum FT(4) levels in moderate dose AP group and high dose AP group were [(9.540 +/- 1.327) fmol/ml] and [(6.509 +/- 1.949) fmol/ml] respectively, which were significantly lower than that [(13.505 +/- 1.276) fmol /ml] in control group (P < 0.05 or P < 0.01). Serum TSH level in high dose AP group was [(1.227 +/- 0.295) mIU/L], which was significantly higher than that [(0.545 +/- 0.282) mIU/L] in control group (P < 0.05). The mRNA expression levels of thyroglobulin (Tg) gene in all groups exposed to AP were significantly lower than that in control group (P < 0.01). The mRNA expression level of thyroperoxidase (TPO) gene in high dose AP group was significantly higher than that in control group (P < 0.05). CONCLUSION: AP can reduce the serum FT(3) and FT(4) levels of rats, increase the serum TSH level of rats and decrease obviously the mRNA expression levels of Tg and TPO genes. In addition, high iodine can reduce the toxic effects of AP on thyroid gland of rats to some extent. PMID- 21619834 TI - [The study of effects of pirfenidone on the pulmonary fibrosis induced by paraquat in mice]. AB - OBJECTIVE: To study the curative effects of pirfenidone (PF) on pulmonary fibrosis induced by paraquat (PQ) in mice and to provide the theoretical basis for clinical treatment. METHODS: Ninety adult healthy male ICR mice were randomly divided into six groups: control group, PQ group, 2 mg/kg Dexamethasone group, 25 mg/kg PF group, 50 mg/kg PF group and 100 mg/kg PF group, there were 15 mice in each group. The corresponding volume of normal saline was given to the each mouse in control group according to the weight, after 2 h 0.1% CMC was given to the each mouse of control group one time by intragastric administration, then the CMC was administrated at regular time until sacrifice. All mice for other 5 groups were exposed to 100 mg/kg PQ by intragastric administration. At 2 h after exposure to PQ, 0.02 ml/10 g dexamethasone and 25, 50, 100 mg/kg PF were given to mice for dexamethasone group and for 3 PF groups by intragastric administration each day for 49 days, respectively. The lung coefficient was calculated and pathological changes of lung tissue were observed by HE staining for each mouse. The hydroxyproline (HYP) level in lung tissue was measured for each mouse. The mRNA level of and the protein level of TGF-beta(1) in lung tissue for each mouse were determined, and the protein level of TGF-beta(1) in the bronchus-alveolus lavage fluid (BALF) of each mouse was detected. RESULTS: The survival rates on the 3rd day in PQ group, 3 PF groups and dexamethasone group were 53.33%, 46.67%, 73.33%, 86.67% and 80%, respectively. The survival rates on the 3rd day in dexamethasone group, 50 mg/kg and 100 mg/kg PF groups were significantly higher than those of PQ group and 25 mg/kg PF group (P < 0.05). The lung coefficients of 3 PF groups were significantly lower than that of the PQ group (P < 0.05). The lung tissue HYP levels of dexamethasone group and 3 PF groups were 50.95 +/- 11.65, 44.52 +/- 9.48, 43.27 +/- 6.01 and 40.82 +/- 5.90 mg/g respectively, which were significantly lower than that (74.27 +/- 3.68) of PQ group (P < 0.01). The TGF-beta(1) protein levels of BALF in dexamethasone group, 50 and 100 mg/kg PF groups were 22.03 +/- 7.27, 27.75 +/- 5.84 and 21.31 +/- 6.82 ng/ml respectively, which were significantly lower than that (52.52 +/- 15.51) ng/ml of PQ group (P < 0.01) The expression level of TGF-beta(1) mRNA in 100 mg/kg PF group decreased significantly, as compared with PQ group (P < 0.01). CONCLUSION: PF could reduce the collagen deposition and pulmonary fibrosis induced by PQ in mice lungs. PMID- 21619835 TI - [Effects of low level manganese exposure on the serum neuroendocrine hormones in the welders]. AB - OBJECTIVE: To study the effects of low level manganese (Mn) exposure on the serum neuroendocrine hormones levels of the welders. METHODS: The exposure group consisted of 41 male welders, 40 male workers without exposing to harmful agents served as controls. The serum contents of prolactin (PRL), luteinizing hormone (LH), follicle stimulating hormone (FSH), testosterone (TST) and thyroid stimulating hormone (TSH) of 81 subjects were detected by chemiluminescence immunoassay. RESULTS: The geometric mean value of airborne Mn concentrations was 0.03 mg/m(3) (0.003 - 0.519 mg/m(3)) in the welding circumstances. The levels of Mn in red blood cells (RBCs) and urinary Mn of the exposure group were significantly higher than those of control group (P < 0.01). The contents of serum LH and TSH of the exposure group were 2.89 +/- 0.69 mIU/ml and 1.45 +/- 0.56 uIU/ml, which were significantly lower than those (3.82 +/- 1.61 mIU/ml and 2.19 +/- 1.28 uIU/ml) of control group (P < 0.01). The serum contents of LH, FSH and TSH of the group exposed to Mn for < 5 years were significantly lower than those of the control group, The serum TST level of the group exposed to Mn for < 5 years was significantly higher than those of the control group and group exposed to Mn for 5 ~ years, the serum FSH level of the group exposed to Mn for < 5 years was significantly lower than that of the group exposed to Mn for 10 years (P < 0.05 or P < 0.01). The serum contents of LH and TSH of the group exposed to Mn for 5 ~ years were significantly lower than those of the control group (P < 0.05 or P < 0.01). The serum contents of PRL, LH and TSH of the group exposed to Mn for 10 years were significantly lower than those of the control group (P < 0.05). There was negative correlation between blood (RBC) Mn and urinary Mn (r = 0.310, P < 0.05), also there was negative correlation between serum PRL and serum TST (r = -0.409, P < 0.01), the positive correlation between serum LH and serum FSH was observed (r = 0.361, P < 0.05). CONCLUSION: The results of present study showed that the long exposure to low level of Mn may decrease the levels of serum PRL, LH and TSH in workers occupationally exposed to Mn, which can influence the metabolism of neuroendocrine hormones to certain extent. PMID- 21619836 TI - [The curative effects of different drugs on liver cell damage of rats induced by acute nickel carbonyl poisoning]. AB - OBJECTIVE: To assess the curative effects of different drugs on liver cell damage of rats induced by acute nickel carbonyl poisoning. METHODS: In present study 220 SD rats were divided into control group (10 rats), carbonyl nickel group (10 rats), 20 mg/kg methylprednisolone group (40 rats), 100 mg/kg DDC group (40 rats), 10 umol/kg sodium selenite group (40 rats), 0.25 ml shenfuhuiyangtang group (40 rats) and 20 mg/kg methylprednisolone with 100 mg/kg DDC group (40 rats). All rats except for control group inhaled passively 250 mg/m(3) carbonyl nickel for 30 minutes. At 4h and 30h after exposure, the drugs were given intraperitoneally to the rats. On the 3rd and 7th days after exposure, the liver samples were taken from 10 rats each group. The DNA damage of liver cells was detected using comet assay, the ultrastructure changes in liver cells were examined under an electronmicroscope. RESULTS: Compared to carbonyl nickel group, the tail lengths of liver cells in 5 groups administrated at 4 h or 30 h and tested on the 3rd or 7th day after exposure decreased significantly (P < 0.05). Compared to the control group, the tail lengths of liver cells in sodium selenite and shenfuhuiyangtang groups administrated at 4h after exposure or sodium selenite, shenfuhuiyangtang and methylprednisolone with DDC groups administrated at 30h after exposure increased significantly (P < 0.05 or P < 0.01), when tested on the 3rd day after exposure. Except from methylprednisolone sub-group administrated at 4h and tested on the 7th day after exposure, the tail lengths of liver cells in other groups administrated at 4 h or 30 h and tested on the 7th day after exposure increased significantly (P < 0.05). Compared to carbonyl nickel group, the Olive moment of liver cells in 5 groups administrated at 4 h or 30 h tested on the 3rd or 7th day after exposure decreased significantly (P < 0.05 or P < 0.01). Compared to the control group, the Olive moment of liver cells in following groups (selenite and shenfuhuiyangtang groups administrated at 4 h or 30 h and tested on the 3rd or 7th day after exposure, DDC group administrated at 4 h or 30 h and tested on the 7th day after exposure, DDC group administrated at 30h and tested on the 3rd day after exposure, and methylprednisolone with DDC group administrated at 30 h and tested on the 7th day after exposure) increased significantly (P < 0.05 or P < 0.01). As compared with carbonyl nickel group, the ultrastructure observation indicated that the nucleus and other organelles of liver cells in methylprednisolone, DDC and methylprednisolone with DDC groups administrated at 4h and tested on the 3rd day were access to normal levels. CONCLUSION: The results of present study showed that methylprednisolone, DDC and methylprednisolone with DDC could improve obviously the repair of rat liver cell damage induced by acute carbonyl nickel poisoning, and the curative effects of early treatment were better than those of later treatment. PMID- 21619837 TI - [The characteristics of type I, III collagen and LN in pulmonary fibrosis induced by uranium ore dust in rats]. AB - OBJECTIVE: To explore the characteristics of LN and type I, III collagen in pulmonary fibrosis induced by uranium ore dust in rats. METHODS: 60 adult Wistar rats were divided randomly into two groups, control group (30 rats) and uranium ore dust group (30 rats). Non-exposed intratracheal instillation method was used. Uranium ore dust group was exposed 20 mg/ml uranium ore dust suspension 1ml per rat, meanwhile control group was exposed normal saline 1ml per rat. Post-exposed the 7, 14, 21, 30 and 60 d, 6 rats in each group were killed randomly, lung tissue were collected. The pathological changes in lung tissue were observed by microscope using HE staining, the collagen I and III in lungs were observed by polarizing microscope using Biebrich scarlet staining. The expression of LN protein in lung tissue was observed by immunohistochemistry-SP. RESULTS: During lung fibrosis, a large amount of the proliferated I and III collagen in lungs were observed. Post-exposure to uranium ore dust, the characteristics in proliferated collagen in lungs were type I collagen deposited in lung interstitium mainly in the early stage. The area percentage of collagen I and III was increased significantly at 7, 14, 21, 30 and 60d in the experimental group as compared with that in the control group (P < 0.05 or P < 0.01). The over expression of LN in the lung tissue were observed. The expression of LN was distributed in the lung tissue as thickening of the linear or cluster. The integral optical density of LN was increased significantly at 21, 30 and 60 d in the experimental group as compared with that in the control group (P < 0.05 or P < 0.01). CONCLUSIONS: After exposure to uranium ore dust, the characteristics in proliferated collagen in lungs are the type of I collagen deposited in lung interstitium mainly in the early stage, while the type of III collagen increase significantly at the later period. The overexpression of LN exists in the process of pulmonary fibrosis. It suggests that LN has a role effect in the process of pulmonary fibrosis. PMID- 21619838 TI - [Risk factors analysis of low back pain among workers in a foundry factory of the automobile company]. AB - OBJECTIVE: To study the main risk factors of low back pain of workers ina foundry factory of the automobile company using cross sectional epidemiological investigation, and to provide scientific base for preventing the disorder. METHODS: The low back pain and work loads of 1340 workers in a foundry factory of the automobile company were investigated using questionnaire, and logistic regression analysis was used to analyze the risk factors. RESULTS: The one-year morbidity of low back pain in workers was 58.9% the morbidities of low back pain in workers engaged in foundry, transportation and modeling were 64.6%, 64.6% and 62.5%, respectively. The lifting with squat postures, bending trunk heavily, bending trunk with twisting and moving the heavy objects were found to be the most dominant risk factors for low-back pain, the OR values were 2.085, 1.961, 1.967 and 1.956, respectively. The distributions of risk factors were different among the different jobs. The logistic regression analysis showed that moving the heavy objects, lifting with squat postures, bending trunk heavily, bending trunk with twisting existed simultaneously, also the work years and gender were the risk factors. CONCLUSION: The manual moving heavy objects, awkward working posture or both were the most important risk factors for low-back pain. The intervene ergonomic study should be performed in future to reduce the morbidity of low-back pain. PMID- 21619839 TI - [A field assessment on the risk factors of musculoskeletal disorders]. AB - OBJECTIVE: To investigate the usability of quick exposure check (Quick Exposure Check, QEC) for the field assessment of occupational musculoskeletal disorder risk factors. METHOD: In the shipyard and automobile manufacturing plants, QEC was used to observe the operations among workers with different jobs and to assess the work loads of workers. On the basis of results, the reliability of QEC was evaluated, and the correlation between QEC scores and morbidities of musculoskeletal disorders in workers was analyzed. RESULTS: The inter-observer reliability (ICC) was in the range from 0.737 to 1.000, and intra-observer reliability (Spearman coefficient) was from 0.605 to 1.000. The order of exposure levels to risk factors of workers engaged in different jobs (QEC scores) in the shipyard factory was plumbers > assemblers > welders; The order of exposure levels to risk factors of workers engaged in different jobs (QEC scores) in the automobile factory was welders > punching workers > machinists > casters > assemblers. In different body parts, the exposure level at back and neck parts was the highest and the exposure level at the shoulder and wrist parts was the second. The regression analysis between QEC scores of body parts and the morbidities of musculoskeletal disorders showed that there was a good correlation between exposure levels and morbidities, the coefficients (r(2)) at the shoulder, wrist, and back (static work) were 0.670, 0.740 and 0.958, respectively (P < 0.05). CONCLUSION: The QEC method is suitable and reliable as demonstrated by the field assessment on the exposure to risk factors in shipyard and automobile workers, and its results is correlated closely to the disease prevalence. PMID- 21619840 TI - [Effects of SOD2 and its C47T mutation on oxidative injury in cochlea hair cell]. AB - OBJECTIVE: To study the effect of SOD2 and its C47T mutation on oxidative injury in cochlea hair cells. METHODS: HEI-OC1 cells were transfected with the SOD2 of Ala16 and Vla16. Cells' proliferation ability was determined by MTT assay. The intracellular SOD2 activities were detected by xanthine oxidase method. Intracellular ROS were determined by DCFH-DA after exposure to 100 umol/L t-BHP and the early apoptotic and necrotic rate or late apoptotic rate were quantified by flow cytometry (FCM) using Annexin V/PI double staining. RESULTS: MTT method showed the transfection of SOD2 gene and empty plasmid did not affect the proliferation capacity. SOD2 vitality in Ala(16) and Val(16) SOD2 transfected cells increased 2.51 and 2.71 times respectively (P < 0.01), but the difference between the two transfection groups was not statistically significant (P > 0.05). After exposed to t-BHP, the majority of the untransfected and empty plasmid transfected cells sent '++' class bright fluorescence, while in Ala(16) and Val(16) SOD2 transfected groups, only about half cells sent '+/-' ~ '+' level fuzzy fluorescence. determination of FCM suggested the early apoptotic and necrotic rate or late apoptotic rate decreased after SOD2 transfection (P < 0.01), but the difference between the two genotypes of SOD2 was not statistically significant (P > 0.05). CONCLUSION: High expression of SOD2 below 3.71 times can reduce intracellular ROS level in HEI-OC1 cells, while SOD2 C47T mutation had no effect on them. SOD2 can be considered as NIHL susceptibility gene and its rs4880 SNP may be not directly related to NIHL genetic susceptibility. PMID- 21619841 TI - [Clinical significance of 5-HT and DA levels in serum and cerebrospinal fluid of the patients with delayed encephalopathy after acute carbon monoxide poisoning]. AB - OBJECTIVE: To explore the changes and the clinical significance of 5 hydroxytryptamine (5-HT), dopamine (DA) levels in serum and cerebrospinal fluid (CSF) of patients with delayed encephalopathy (DEACMP) after acute carbon monoxide poisoning. METHODS: The dynamic detection of 5-HT and DA levels in serum and CSF from 42 patients with DEACMP was performed with high performance liquid chromatography (HPLC). The condition changes of patients with DEACMP were analyzed with three types of scales: the activity of daily living scale (ADL), information memory concentration test (IMCT) and Hasegawa's dementia scale (HDS); these changes were compared with those from 38 other encephalopathy patients and 38 non-encephalopathy patients, respectively. RESULTS: Before treatment, the serum 5-HT and DA levels [(662.61 +/- 178.50) and (155.74 +/- 60.32) nmol/L, respectively] of DEACMP group were both significantly lower than those [(914.08 +/- 198.04) and (225.70 +/- 48.53) nmol/L] of non-encephalopathy group (P < 0.05); the serum DA level of DEACMP group was also significantly lower than that [(243.57 +/- 66.94) nmol/L] of other encephalopathy group (P < 0.05); the serum 5 HT level of DEACMP group was not significantly different from that [(729.54 +/- 299.87) nmol/L] of other encephalopathy group (P > 0.05). After treatment, the serum 5-HT and DA levels [(714.08 +/- 170.47) and (192.18 +/- 33.07 nmol/L, respectively)] of DEACMP group elevated to various extent, but only serum DA level was significantly higher than that before treatment (P < 0.05). Before treatment, the CSF 5-HT and DA levels of DEACMP group were significantly lower than those of non-encephalopathy group and those of other encephalopathy group (P < 0.05). After treatment, the CSF 5-HT level (232.44 +/- 54.28 nmol/L) was similar to normal level and significantly higher than that before treatment (P < 0.05); the CSF DA level [(56.83 +/- 12.85) nmol/L] of DEACMP group increased only slightly (P > 0.05). In DEACMP group, ADL score (50.64 +/- 7.23), HDS score (8.55 +/- 8.08) and IMCT score (4.95 +/- 7.30) before treatment were significantly different from those (8.5 +/- 8.08, 4.95 +/- 7.30 and 15.64 +/- 10.90) after treatment (P < 0.01). In DEACMP group, there wasa negative correlation between DA level changes and HDS score changes, when the DA levels and HDS scores before treatment were compared with those after treatment (P < 0.05). CONCLUSION: The dynamic changes of 5-HT and DA levels in serum and CSF of patients with DEACMP consisted basically with the patient's condition change. The dynamically detected 5-HT and DA levels can be used as the biological indicators to reflect the condition change and treatment effects of DEACMP patients. PMID- 21619842 TI - [The changes of blood neurotransmitter levels in workers occupationally exposed to ethylbenzene]. AB - OBJECTIVE: To explore the effects of occupational ethylbenzene exposure on blood neurotransmitter levels in population. METHODS: The exposure group consisted of 246 workers occupationally exposed to ethylbenzene and the control group was composed of 122 staffs from the offices. The basic information on ethylbenzene exposure was collected by the questionnaire. The mandelic acid (MA) and phenylglyoxylic acid (PGA) in the post-working urine were measured using the high performance liquid chromatography. The levels of gamma-aminobutyric acid (GABA), dopamine (DA) and acetylcholinesterase (AchE) activity were detected by reversed phase high performance liquid chromatography, spectrofluorometry and DTNB method, respectively. The blood biochemical indexes: alanine transaminase (ALT), aspartate aminotransferase (AST), total protein (TP), albumin (ALB), alkaline phosphatase (ALP), total bilirubin (TBIL) were examined. Also the hematologic indexes: red blood cell (RBC), white blood cell (WBC), hemoglobin (HGB) and platelet (PLT) were determined. RESULTS: The levels of MA, PGA and MA+PGA of urine in the exposed group were significantly higher than those in the control group (P < 0.05). There were no significant differences of the biochemical indexes (AST, ALT, TP, ALB, BUN, Cr, ALP and TBIL), hematologic indexes (WBC, RBC, Hb and PLT) and serum GABA between the exposure group and the control group (P > 0.05). But the serum DA [(0.21 +/- 0.011) mg/L] and AChE levels [(0.321 +/- 0.066) U/L] in the exposure group were significantly lower than those in the control group [(0.25 +/- 0.015) mg/L, (0.583 +/- 0.125) U/L], respectively (P < 0.05). CONCLUSION: MA and PGA in urine can serve as the biomarkers of internal exposure dose. Before the obvious changes of biochemical indexes and hematologic indexes appear, the exposure to ethylbenzene can influence the blood neurotransmitter levels in workers exposed to ethylbenzene. PMID- 21619843 TI - [The neurobehavioral effects of population occupationally exposed to ethylbenzene]. AB - OBJECTIVE: To evaluate the influence of ethylbenzene on the neurobehavior of occupationally exposed workers. METHODS: The exposure group consisted of 246 workers occupationally exposed to ethylbenzene and 172 staffs from the offices served as controls. The basic information on ethylbenzene exposure was collected by the questionnaire. The nervous behavior and function of workers were evaluated by Neurobehavioral Core Test Battery (NCTB). RESULTS: There were no differences of the scores for four emotional states (tension, depression, angry and bewilderment) between exposure group and control group (P > 0.05). The score of emotion (vigor) in exposure group was significantly lower than that in control group (P < 0.05), but the fatigue score in exposure group was significantly higher than that in control group (P < 0.05). The score of mean reaction time in exposure group was significantly higher than that in control group (P < 0.05), the scores of digital span, manual dexterity, visual retention and target tracking in exposure group were significantly lower than those in control group (P < 0.05). The exposure group was divided into 5 sub-groups, according to working duration. There were no differences for the scores of visual retention and target tracking among 5 sub-groups (P > 0.05). The scores of five emotional states (tension, depression, angry, fatigue and bewilderment) in 3 sub-groups exposed to ethylbenzene for 3 ~, 4 ~ and 5 ~ years were significantly higher than those in 2 sub-groups exposed to ethylbenzene for 0 ~ and 2 ~ years (P < 0.05). The scores of digital span in 2 sub-groups exposed to ethylbenzene for 3 ~ or 4 ~ years and the scores of manual dexterity and digital symbol in 3 sub-groups exposed to ethylbenzene for 3 ~, 4 ~ and 5 ~ years were significantly lower than those in 2 sub-groups exposed to ethylbenzene for 0 ~ and 2 ~ years (P < 0.05). CONCLUSION: Ethylbenzene can depress the neurobehavioral functions of exposed workers. The neurobehavioral functions of workers exposed to ethylbenzene for 3 years changed significantly. The workers exposed to ethylbenzene for 3 years may be the susceptible population of neurobehavioral function impairment. PMID- 21619844 TI - [Effects on serum myelin proteins of n-hexane exposure]. AB - OBJECTIVE: Exploring the effects of n-hexane on expression of serum myelin proteins in occupational exposure workers, and finding the early biomarker of n hexane exposure. METHODS: In the study, 373 subjects were recruited, 269 exposure workers (work experience of more than1 year) and 104 non-exposure workers were selected. Firstly examined the level of urinary 2,5-hexanedione in the two groups, based on urinary 2,5-hexanedione biological limit value (4 mg/L), the exposed group was divided into high-exposed group and low-exposed group. And then collected blood samples and extracted serum. Human peripheral myelin protein zero (P0) antibody (IgG, IgM) and human peripheral myelin protein two (P2) antibody (IgG, IgM) analysis was performed according to ELISA kit. RESULTS: The concentration of urinary 2,5-hexanedione in the exposed group was (3.10 +/- 1.35) mg/L. The level of P0 antibody (IgG, IgM) and P2 antibody (IgG, IgM) in the high exposed group and low-exposed group were both higher than that in the controls (P < 0.01). CONCLUSION: P0 antibody and P2 antibody could be used as the early biomarkers of n-hexane exposure, which not only evaluate the occupational hazards in the early, but also provide the policy maker with scientific evidence. PMID- 21619845 TI - [The curative effects of transmetil on Amanita verna poisoning]. AB - OBJECTIVE: To observe the curative effects of transmetil on Amanita verna poisoning. METHODS: Twelve cases with Amanita verna poisoning were reviewed. The patients were divided into 2 groups according to usage of transmetil: Group A was treated with traditional protocol (gastric lavage, catharsis, rehydration, diuresis, anti-infection and hemodialysis), Group B was treated with traditional protocol combined with transmetil. The liver function changes on the 1st, 3rd, 5th and 7th day after poisoning and the mortality were compared between 2 groups. RESULTS: Two cases in group A (6 patients) died. The mortality of group A was 33.3%. The AST levels continued to increase on the 3rd and 5th day, but decreased on the 7th day. TBIL continued to increased on the 1st, 3rd, 5th and 7th day. None in group B died. The TBIL level dropped at 7 d 5 patients showed an increase in ALT at 7 d and 3 patients showed a decrease in AST at 7 d. CONCLUSION: Transmetil may play an important role in reducing the mortality of Amanita verna poisoning. PMID- 21619846 TI - [Development of the certified reference material of mercury in lyophilized human urine]. AB - OBJECTIVE: To develop the certified reference material of mercury in lyophilized human urine. METHODS: Human urine samples from normal level mercury districts were filtered, homogenized, dispensed, lyophilized and radio-sterilized. Homogeneity test, stability inspection and certification were conducted using a atom fluorescence spectrophotometric method. The physical and chemical stability of the certified reference material were assessed for 18 months. The certified values are based on analysis made by three independent laboratories. RESULTS: The certified values are as follows: low level was (35.6 +/- 2.1) ug/L, high level was (50.5 +/- 3.0) ug/L. CONCLUSION: The certified reference material of mercury in lyophilized human urine in this research reached the national certified reference material requirements and could be used for the quality control. PMID- 21619847 TI - [Determination of trimethyltin chloride in urine by headspace-gas chromatography]. AB - OBJECTIVE: To establish a detection method for trimethyltin chloride in urine by the Head space-GC. METHOD: After derivatizing trimethyltin chloride, the urines was separated by the head space-gc, and then the trimethyltin chloride detected qualitatively and quantificationally. RESULTS: In the concentration range of 0.02 ~ 0.40 mg/L urinary trimethyltin chloride, showed a quadratic, r = 0.9992, detection limit was 0.005 mg/L, the relative standard deviation was 1.9% ~ 2.5%, recovery was 92.0% to 100%, the urine samples can be saved at least 90 days in 18 degrees C refrigerator. CONCLUSION: The instrument, reagents involved in the detection require low, the operations to processing samples are simple, high sensitivity, less interference, good reproducibility, and suitable for quantitative and qualitative analysis, convenient to promotion. PMID- 21619848 TI - [Determination of phosphoric acid in the air of workplace by ion chromatography]. AB - OBJECTIVE: To develop a method to determine phosphoric acid in the air of workplace by ion chromatography. METHODS: Phosphoric acid was collected by millipore filter and washed by deionized water then detected by ion chromatography. RESULTS: Linearity range of test was 0 ~ 20 ug/ml, relative standard deviation (RSD) was 1.95% ~ 3.31%, the elution efficiency was 103.0% ~ 109.6%, determination limit was 0.1 ug/ml (when sample size was 20.01) concentration limit was 0.01 mg/m(3) (when the collected air was 75 L). CONCLUSION: This method is convenient for air collection, simple, with high sensitivity and good precision, is a good method for determination of phosphoric acid in the air of workplace. PMID- 21619849 TI - [Simultaneous determination of seven chemicals of halogenated alkanes and aromatic hydrocarbons in the air of workplace by gas chromatography]. AB - OBJECTIVE: To establish a gas chromatographic method for determination of halogenated alkanes and aromatic hydrocarbons including trichloromethane, 1,2 dichloroethane, carbon tetrachloride, benzene, toluene, ethylbenzene and xylene in the air of workplace. METHODS: After the air samples collected with activated carbon tubes and desorbed with CS(2), the target toxicants were separated with FFAP capillary columns and detected with flame ionization detector. RESULTS: The coefficient of correlation was above 0.999 and the lowest detectable concentrations were 0.2 ~ 3.6 mg/m(3) with the RSD of 1.2% ~ 4.6%. The desorption efficiencies was 94.9% ~ 100.7%. CONCLUSION: The method shows lower detection limit, high accuracy and precision. It is feasible for determination of the seven halogenated alkanes and aromatic hydrocarbons in the air of workplace. PMID- 21619850 TI - [Determination of phosphorus oxychloride in the air of workplace by ion chromatography]. AB - OBJECTIVE: To establish a ion chromatography method for determination of phosphorus oxychloride in the air of workplace. METHOD: The phosphorus oxychloride in the air of workplace was collected by absorb liquid and turned into hydrochloric acid, then separated in column and detected with conductivity detector, qualified by elution time and quantified by peak height or peak area. RESULTS: The linear range of phosphorus oxychloride in air of workplace was 0.72 ~ 5.76 ug/ml with its correlation coefficient 0.9999. The detecting limit of the method was 0.12 ug/ml. The smallest detecting concentration of the method was 0.08 mg/m(3) for 15 L sampling air. Relative standard deviation was 3.3% ~ 6.2% and the recovery was 97.8% ~ 103.8%. The sample could be resaved at room temperature at least for seven days. CONCLUSION: The indicators of the method correspond GBZ/T 210.4-2008"Guide for establishing occupational health standards Part 4: Determination methods of air chemicals in workplace". It is a good method to determine phosphorus oxychloride in the air of workplace. PMID- 21619851 TI - [Determination of tetrahydrofuran in urine by headspace solid-phase microextraction and gas chromatography]. AB - OBJECTIVE: Headspace solid-phase microextraction (HS-SPME) was used pre concentration procedure for the determination of tetrahydrofuran in urine by gas chromatography with hydrogen flame detector. METHODS: Several parameters controlling SPME was studied and optimised: SPME fiber, extraction time and extraction temperature, desorption time and desorption temperature. RESULTS: Under optimal conditions, the correlation coefficient was 0.9998 and good recoveries (range from 93.0% ~ 100.8%) were achieved, the detection limit was 0.5 ug/L. CONCLUSION: The method can be applied to the determination of trace amount of tetrahydrofuran in urine. PMID- 21619852 TI - [Determination of o-chlorostyrene in the air of workplace by gas chromatography]. AB - OBJECTIVE: To establish a rapid gas chromatographic method for determination of o chlorostyrene in the air of workplace. METHOD: The air samples were collected by syringes, injected directly to the GC system, and then separated by a FFAP capillary column (30 m * 0.53 mm * 0.25 um), finally determined by a Flame Ionization Detector. RESULTS: A good linear correlation was showed within a range of 0 ~ 1200 ug/L, with regression formula Y = 14 030 + 7 207X (r = 0.9999). The air sample could be stably stored in the syringe for 5 hrs. The relative standard deviation (RSD) of repeated injection of o-chlorostyrene standard solutions at three different concentration by six times was 1.28% ~ 1.97%. The minimum detectable concentration was calculated to be 5.2 mg/m(3). Other coexistent violative organic compounds such as styrene, p-chlorostyrene, and m-chlorostyrene didn't interfere with the determination under the experimental conditions of this method. CONCLUSION: This method meets the requirement of "Guide for establishing occupational health standards-Part 4: Determination methods of air chemicals in workplace". It is applicable for determination of o-chlorostyrene in the air of workplace. PMID- 21619853 TI - Calibration of the Ph. Eur. Biological Reference Preparation (BRP) for tetanus vaccine (adsorbed) batch 3. AB - A joint collaborative study was organised by the European Directorate for the Quality of Medicines & HealthCare (EDQM) and the World Health Organization (WHO)/National Institute for Biological Standards and Control (NIBSC) to establish replacement batches for the European Pharmacopoeia (Ph. Eur.) Tetanus Vaccine (adsorbed) Biological Reference Preparation (BRP) batch 2 and for the WHO 3rd International Standard (IS) for Tetanus toxoid (adsorbed). Two freeze-dried stabilised tetanus vaccine (adsorbed) candidate preparations (Preparation A, 08/218 and Preparation B, 08/102) were calibrated against the current 3rd IS/BRP batch 2 (Preparation C) using challenge methods in guinea pigs and mice as described in the Ph. Eur. general chapter 2.7.8. Assay of tetanus vaccine (adsorbed). They were also assayed by serology methods. The WHO 2nd IS for Tetanus toxoid adsorbed (TEXA-2) was additionally included in the sample panel as Preparation D. Thirty-four laboratories (regulatory organisations and manufacturers) from 22 countries participated in the collaborative study. The majority of participants performed 2 independent challenge tests. Nine laboratories performed challenge assays in guinea pigs and 30 laboratories performed challenge assays in mice. Eight laboratories performed serology in guinea pigs and 1 laboratory performed serology in mice. For Preparation A, the geometric mean (GM) potency estimate (with 95 % confidence interval (CI)) in guinea pigs for all laboratories that provided valid results (n = 6) was 488.5 (354.2-673.6) IU/ampoule. For valid mouse assays (n = 25) the GM potency (with 95 % CI) was 259.8 (223.5-302.0) IU/ampoule. The inter-laboratory geometric coefficient of variation (GCV) was 36 % for guinea pig assays and 45 % for mouse assays. This compared favourably with the calibration of the 3rd IS/BRP batch 2 where the inter-laboratory GCV was 36 % and 42 % in guinea pigs and mice, respectively. For Preparation B, the GM potency estimate (with 95 % CI) in guinea pigs for all laboratories that provided valid results (n = 6) was 107.9 (64.1 181.7) IU/ampoule. For valid mouse assays (n = 24) the GM potency (with 95 % CI) was 147.9 (126.3-173.1) IU/ampoule. The inter-laboratory GCV was 64.3 % for guinea pig assays and 45.2 % for mouse assays. From the collaborative study, Preparation A appeared more suitable to be the replacement Ph. Eur. BRP as it is similar to the Tetanus vaccine (adsorbed) BRP batch 2, except for nature of the stabiliser. Preparation A was confirmed to have higher potency, readily detectable tetanus toxoid, and confirmed satisfactory stability and performance in challenge assays. Preparation A was adopted in January 2011 by the Ph. Eur. Commission as the Tetanus vaccine (adsorbed) BRP batch 3, with assigned potencies of 490 IU/ampoule in the guinea pig challenge assay and of 260 IU/ampoule in the mouse challenge assay. The same Preparation A was adopted in October 2010 as the WHO 4th IS for Tetanus toxoid (adsorbed), with the assigned activity of 490 IU/ampoule from guinea pig challenge assays. A follow-up study (reporting study) was organised by the EDQM to assess the impact of the potency assigned to the BRP batch 3 for mouse challenge assays on the outcome of batch release testing in Europe. Eight laboratories including official medicines control laboratories (OMCLs) and manufacturers reported the results of their routine testing, using the BRP batch 3 in addition to their regular reference preparation. For each tested product, participants calculated the potency relative to their routine reference and relative to the BRP batch 3. No common sample panel was distributed to participants. In total, data on 40 batches of different marketed tetanus vaccines were reported. Overall, a good concordance was observed between the potencies calculated relative to the BRP batch 2 and relative to the BRP batch 3. On average, the potency estimates were 10 % lower when expressed relative to the BRP batch 3. Cases of discrepant decisions for batch release were very limited and affected mainly batches with specifications close to the pharmacopoeial requirements. The reasons for differences in estimated potencies are discussed. The study showed that the use of the BRP batch 3 with an assigned potency of 260 IU/ampoule does not result in substantial change in the potency of different marketed products. This confirmed that the mouse challenge potency value assigned to the BRP batch 3 is suitable. PMID- 21619854 TI - Collaborative study on influenza vaccine clinical trial serology - part 1: CHMP compliance study. AB - The Quality of Medicines & HealthCare (EDQM, Council of Europe) and the European Union (EU) Commission to evaluate the reproducibility of clinical serology results for seasonal influenza vaccines and to assess the impact of technical differences between laboratories on the compliance with the Committee for Human Medicinal Products (CHMP) criteria set by the European Medicines Agency (EMA). The study was run in 2 phases. The present article reports the 1st phase of the study, which aimed at evaluating the variability of the results obtained by 11 laboratories (5 national control laboratories and 6 influenza vaccine manufacturers) using their routine haemagglutination inhibition (HI) assay to test a common panel of clinical trial sera. The results confirmed the limited inter-laboratory reproducibility of the HI testing of influenza vaccine clinical trial samples. In some cases a good agreement was found between laboratories, while a systematic bias or a random scatter of results was observed in other cases. Analysis of estimated systematic bias confirmed that differences between laboratories can be significant (up to 16-fold) in some cases. Correction for this bias resulted in limited improvement. Differences between laboratories were found to result in discrepant decisions on marketing acceptance of vaccines or to decisions based on compliance to different criteria. The study showed that the seroconversion (SC) and mean fold increase (MFI) criteria are more robust against systematic over- or under-estimation of titres whereas the protection rate (PR) is very sensitive to this effect. The fundamental issues with the PR criteria are discussed. PMID- 21619855 TI - Collaborative study on influenza vaccine clinical trial serology - part 2: reproducibility study. AB - A collaborative study was run by the Biological Standardisation Programme (BSP) under the aegis of the European Directorate for the Quality of Medicines & HealthCare (EDQM, Council of Europe) and the European Union (EU) Commission, to address the issue of the poor standardisation of serological assays used for the evaluation of seasonal influenza vaccines in Europe. The Phase 1 of the study focused on the compliance to Committee for Human Medicinal Products (CHMP) criteria by 6 manufacturers and 5 public laboratories. It confirmed the poor inter-laboratory correlation of haemagglutination inhibition (HI) test results. Phase 2 consisted in a reproducibility study examining the impact of extended method standardisation and the use of reference sera on inter-laboratory variation. Six manufacturers and 5 public laboratories contributed HI results, while the 5 public laboratories also performed single radial haemolysis (SRH) tests on the same sample panels. Results showed that method standardisation failed to significantly improve the inter-laboratory variation. Correction for pre-vaccination titres (Beyer correction) was found to have limited effect to improve the bias constituted by the Protection Rate (PR) criterion. The reasons underlying the difficulty in standardisation of HI and SRH tests are discussed and improved approaches for the compliance testing to CHMP criteria are suggested. PMID- 21619856 TI - Viral safety in homoeopathic medicinal products. AB - To guarantee the safety of medicinal products as regards infectious agents, numerous national guidelines and recommendations have in recent years been included in the pharmacopoeia general monographs and have influenced the content of the substance monographs. Although the stipulations of the European Pharmacopoeia set out objectives, there is still a certain scope in how the requirements are implemented. This is reflected in the very different responses in Europe to the problems of safety from infection. Different traditions in the use of homoeopathic and anthroposophic therapy and varying levels of expertise among the regulatory authorities within the European Union have resulted in varying standard of assessment. The aim of this publication is to present a standard form of assessment for medicinal products in these therapeutic systems. Demonstrated hereunder is an approach that can be adopted to ensure that the high safety standard required is met for homoeopathic and anthroposophic medicinal products. PMID- 21619857 TI - An alternative to animal testing in the quality control of erythropoietin. AB - A physico-chemical method has been developed as an alternative to the current bioassay in normocythaemic mice for estimating the biological activity of erythropoietin batches. Capillary zone electrophoresis was used for quantification of the isoforms and their substructures were further elucidated by N-glycan mapping techniques. The analytical study was carried out on a total of 40 batches of epoetin beta which were selected to cover an adequate range of precisely established potency values. The relationship between the biological and chemical parameters was evaluated statistically in order to identify suitable covariates for the prediction of the biological activity. Out of several alternatives, a prediction model which is based on the percentages of isoforms per batch and the degree of sialidation was selected and tested. This model is comparable in terms of accuracy to the established in vivo bioassay, but is far superior in terms of precision. Further advantages of the method are improved animal welfare and savings in time and effort. The question whether the prediction model already meets the requirements for replacing the bioassay according to the ICH guideline Q6B is discussed. PMID- 21619858 TI - How to evaluate the separation efficiency of CZE methods? AB - In trying to estimate the separation efficiency of Capillary Electrophoresis (CE) methods, the resolution (RS), the number of theoretical plates (N) and the peak to-valley ratio (p/v) are often used assessment criteria. This study demonstrates that these criteria are not as suitable to describe the separation efficiency in case of Capillary Zone Electrophoresis (CZE) methods as they are for Liquid Chromatography (LC) methods. The investigations were performed by means of a validated CZE method for the evaluation of tetracyclines and their related substances. Four impurities of tetracycline hydrochloride are described in the European Pharmacopoeia. Three were found in the sample used for our investigations, i.e. epi-tetracycline formed by keto-enol-tautomerism, anhydrotetracyclin and epi-anhydrotetracyline. It could be shown that higher values of these assessment criteria like RS do not necessarily represent better separation. Thus, a discussion on the usefulness of separation selectivity and efficiency as assessment criteria for capillary electrophoresis as well as on the introduction of additional parameters is needed. PMID- 21619859 TI - Schema therapy, motivational interviewing, and collaborative-mapping as treatment for depression among low income, second generation Latinas. AB - US-born Latinos report significantly more depression than foreign-born Latinos in the US, and Latinas have twice the rate of depression than Latino men. The purpose of this pilot study was to test the feasibility of an innovative, short term program of Schema Therapy (ST) combined with Motivational Interviewing (MI) techniques to reduce depression and increase resilience among second generation Latinas of low income in the US. In addition to blending ST and MI strategies with a focus on resilience, a novel technique called collaborative-mapping was a crucial strategy within treatment. Scheduling for sessions was flexible and patients had unlimited cell phone access to the therapist outside of sessions, although few used it. A mixed linear regression model for BDI-II scores of 8 women who completed all eight 2-h sessions demonstrated that the treatment significantly decreased BDI-II scores during the course of treatment (p = .0003); the average decreasing rate in BDI-II scores was 2.8 points per visit. Depression scores remained sub-threshold for 12 months after treatment completion. Resilience scores significantly increased after treatment completion and remained high at all follow-up visits through 1 year (p < .01). Thus, this short term, customized intervention was both feasible and effective in significantly decreasing depression and enhancing resilience for this sample with effects enduring one year after treatment. This study is the first to combine ST and MI in therapy, which resulted in an appealing, desirable, and accessible depression treatment for this severely understudied, underserved sample of low income, second generation Latinas in the US. PMID- 21619860 TI - Neuroprotection in acute cerebral ischemia: can we improve clinical outcomes? PMID- 21619861 TI - Epidemiology of ischaemic stroke and traumatic brain injury. AB - Acquired brain injury, including both Ischaemic stroke (IS) and Traumatic Brain injury (TBI), is one of the most common causes of disability and death in adults. Yet there are vast differences in our knowledge of their epidemiology. While the incidence, case-fatality and risk factors for stroke are well established, work needs to continue particularly in low-income countries, where these data remain sparse; and in relation to specific stroke subtypes such as IS. Similar data regarding the epidemiology of TBI are generally lacking. The majority of TBI incidence studies have focussed on hospital-based samples and there are no established criteria from which to design high quality epidemiological studies. The need to establish such criteria separate from those already available for stroke is suggested given the differing demographic profile of TBI as well as differences in seeking of medical attention for TBI. The immense burden of stroke can be reduced by prevention of modifiable risk factors particularly in developing countries where both changing lifestyle and lack of healthcare resources are contributing to rising stroke incidence and mortality. Similarly, studies to date indicate that TBI incidence can be reduced by addressing modifiable risk factors such as alcohol abuse, risk-taking behaviour and socioeconomic disparities. PMID- 21619862 TI - Acute pathophysiological processes after ischaemic and traumatic brain injury. AB - Ischaemic stroke and brain trauma are among the leading causes of mortality and long-term disability in the western world. Enormous endeavours have been made to elucidate the complex pathophysiology of ischaemic and traumatic brain injury with the intention of developing new therapeutic strategies for patients suffering from these devastating diseases. This article reviews the current knowledge on cascades that are activated after ischaemic and traumatic brain injury and that lead to progression of tissue damage. Main attention will be on pathophysiological events initiated after ischaemic stroke including excitotoxicity, oxidative/nitrosative stress, peri-infarct depolarizations, apoptosis and inflammation. Additionally, specific pathophysiological aspects after traumatic brain injury will be discussed along with their similarities and differences to ischaemic brain injury. This article provides prerequisites for understanding the therapeutic strategies for stroke and trauma patients which are addressed in other articles of this issue. PMID- 21619863 TI - Free radical scavengers and spin traps--therapeutic implications for ischemic stroke. AB - Ischemic stroke comprises a complex cascade of pathophysiological mediators among which reactive oxygen species (ROS) play a pivotal role. Although oxidative stress as one major component contributing to ischemia-reperfusion injury has been thoroughly studied before, efficient treatment options for patients with ischemic stroke have so far not been transferred into clinical practice. In this review, the authors first describe some of the fundamental pathophysiological mechanisms that are involved in ROS generation after cerebral ischemia. Thereafter, antioxidant defense mechanisms and pharmacological manipulation of oxidative stress in various models of experimental cerebral ischemia are reviewed. The authors finally comment on recent clinical studies analyzing the effect of an antioxidative therapy after ischemic stroke and present a short outlook for further studies on ROS-mediated injury after stroke. PMID- 21619864 TI - Preconditioning and postconditioning for neuroprotection: the most recent evidence. AB - Stroke is a leading cause of morbidity and mortality, with perioperative stroke being an important complication in the practice of anaesthesia. Unfortunately, pharmacological treatment options are very limited and often not applicable in the perioperative period. The notion of applying a subtoxic stimulus prior to an otherwise lethal event is termed preconditioning. The main focus of the article is on describing the different concepts of preconditioning, including remote ischaemic preconditioning and anaesthetic preconditioning, as well as postconditioning and summarizing the most recent discoveries in this exciting field. PMID- 21619865 TI - Perioperative neuroprotection. AB - The endpoint of all cerebral injuries like stroke, global cerebral ischemia during cardiac arrest, cardiac, vascular, or brain surgery or head trauma is the inadequate supply of the brain with oxygen and glucose, which triggers a characteristic pathophysiologic cascade leading to neuronal death. Many methods and agents have been investigated to produce neuroprotection from cerebral ischemia along this cascade (e.g., hypothermia, anaesthetics, free radical scavengers, excitatory amino acid antagonists, calcium channel blockers, ionic pump modulators, growth factors, heparinization, antineutrophil/platelet factors, steroids, and gene products). However, essentially none of the pharmacological approaches was identified as useful in humans though most agents have been successfully tested in animal models. Expert opinion suggests that neuroprotective approaches have failed in human trials because there are multiple mechanisms of injury from local and cerebral ischemia. Furthermore, adequate timing might essential because of the temporal sequence of cerebral injury. However, because there are multiple mechanisms of injury, there are most likely also multiple mechanisms of neuroprotection. The most important strategy is profound knowledge on cerebral physiology and homeostasis in health and disease. This review discusses essential physiological mechanisms to warrant adequate supply of glucose and oxygen to the brain. In addition, the influence of potential neuroprotective strategies and agents are reviewed in the perioperative setting. PMID- 21619866 TI - Neuronal injury in chronic CNS inflammation. AB - INTRODUCTION: Multiple sclerosis (MS) is the most common chronic inflammatory disease of the central nervous system which is characterized by inflammatory demyelination and neurodegeneration. Neurological symptoms include sensory disturbances, optic neuritis, limb weakness, ataxia, bladder dysfunction, cognitive deficits and fatigue. PATHOPHYSIOLOGY: The inflammation process with MS is promoted by several inflammatory cytokines produced by the immune cells themselves and local resident cells like activated microglia. Consecutive damaging pathways involve the transmigration of activated B lymphocytes and plasma cells, which synthesize antibodies against the myelin sheath, boost the immune attack, and result in ultimate loss of myelin. Likewise, activated macrophages and microglia are present outside the lesions in the normal-appearing CNS tissue contributing to tissue damage. In parallel to inflammatory demyelination, axonal pathology occurs in the early phase which correlates with the number of infiltrating immune cells, and critically contributes to disease severity. The spectrum of neuronal white matter and cortical damage ranges from direct cell death to subtle neurodegenerative changes such as loss of dendritic ramification and the extent of neuronal damage is regarded as a critical factor for persisting neurological deficits. Under normal conditions, CNS microglia safeguards organ integrity by constantly scanning the tissue and responding rapidly to danger signals. The main task of microglial cells is to encapsulate dangerous foci and remove apoptotic cells and debris to protect the surrounding CNS tissue; this assists with tissue regeneration in toxin-induced demyelination. In the absence of lymphocytic inflammation and in the context of non-autoimmune, pathogen-associated triggered inflammation, microglial cells protect the neuronal compartment. These mechanisms seem to be inverted in MS and other chronic neurodegenerative disorders because activated microglia and peripherally derived macrophages are shifted towards a strongly pro-inflammatory phenotype and produce the proinflammatory cytokines TNF-alpha and interleukin (IL)1-beta, as well as potentially neurotoxic substances including nitric oxide, oxygen radicals and proteolytic enzymes. Microglial silencing reduces clinical severity, demonstrating their active involvement in damage processes and in the immune attack against the CNS. In light of this, it is questionable whether microglia and monocyte-derived macrophages, the very last downstream effector cells in the immune reaction, actually have the capacity to influence their fate. It is more likely that the adaptive immune system orchestrates the attack against CNS cells and drives microglia and macrophages to attack oligodendrocytes and neurons. NEUROPROTECTIVE STRATEGIES: Currently, Glatiramer acetate (GA) and the interferon beta (IFN-beta) variants are established as first-line disease modifying treatments that reduce the relapse rate, ameliorate relapse severity and delay the progression of disability in patients with relapsing-remitting MS. Similarily, sphingosine-1-phosphate (S1P) receptor agonists which influence lymphocyte migration through T cells-trapping in secondary lymphatic organs ameliorates astrogliosis and promotes remyelination by acting on S1P-receptors on astrocytes and oligodendrocytes. Ion channel blockers (e.g. sodium channel blockers), currently used for other indications, are now tested in neurodegenerative diseases to restore intracellular ion homeostasis in neurons. Axonal degeneration was significantly reduced and functional outcome was improved during treatment with Phenytoin, Flecainide and Lamotrigine. Although evidence for a direct protective effect on axons is still missing, additional immune modulatory actions of sodium channel blockers on microglia and macrophages are likely available. In vitro-studies in axons subjected to anoxia in vitro or exposure to elevated levels of nitric oxide (NO) in vivo demonstrated the involvement of a direct effect on axons. As increased intracellular calcium levels contribute to axonal damage through activation of different enzymes such as proteases, blockade of voltage gated calcium channels is another promising target. For example, nitrendipin and bepridil ameliorate axonal loss and clinical symptoms in different models of chronic neurodegeneration. In addition to these exogenous neuroprotective patheways, endogenous neuroprotective mechanisms including neurotrophins, (re)myelination and, neurogenesis support restauration of neuronal integrity. PMID- 21619867 TI - The only evidence based neuroprotective therapy for acute ischemic stroke: thrombolysis. AB - Occlusion of a brain vessel leads to a critical reduction in cerebral perfusion and, within minutes, to ischemic infarction with a central infarct core of irreversibly damaged brain tissue and a more or less large area of hypoperfused but still vital brain tissue (the ischemic penumbra), which can be salvaged by rapid restoration of blood flow. Therefore, the underlying rationale for the introduction and application of thrombolytic agents is the lysis of an obliterating thrombus and thus reestablishment of cerebral blood flow by cerebrovascular recanalization with subsequent reperfusion. After introduction of thrombolytic therapy for the treatment of acute myocardial infarction in the early 1990 s, major trials for the evaluation of this new therapeutic approach to ischemic stroke were initiated. There are in general two strategies in thrombolytic therapy, a local (intraarterial) approach and a systemic (intravenous) application of the thrombolytic agent. Only the latter has been proven to be effective in larger randomized trials and is a Class 1/Level A recommendation in national or international guidelines.(1) This review summarizes the evidence for thrombolytic therapy of acute ischemic stroke with emphasis on progress regarding the approved intravenous treatment. Finally intraarterial approaches as well as combined systemic and interventional therapies are discussed. PMID- 21619868 TI - Erythropoietin as neuroprotective and neuroregenerative treatment strategy: comprehensive overview of 12 years of preclinical and clinical research. AB - Erythropoietin (EPO), originally discovered as hematopoietic growth factor, has direct effects on cells of the nervous system that make it a highly attractive candidate drug for neuroprotection/neuroregeneration. Hardly any other compound has led to so much preclinical work in the field of translational neuroscience than EPO. Almost all of the >180 preclinical studies performed by many independent research groups from all over the world in the last 12 years have yielded positive results on EPO as a neuroprotective drug. The fact that EPO was approved for the treatment of anemia >20 years ago and found to be well tolerated and safe, facilitated the first steps of translation from preclinical findings to the clinic. On the other hand, the same fact, naturally associated with loss of patent protection, hindered to develop EPO as a highly promising therapeutic strategy for application in human brain disease. Therefore, only few clinical neuroprotection studies have been concluded, all with essentially positive and stimulating results, but no further development towards the clinic has occurred thus far. This article reviews the preclinical and clinical work on EPO for the indications neuroprotection/neuroregeneration and cognition, and hopefully will stimulate new endeavours promoting development of EPO for the treatment of human brain diseases. PMID- 21619869 TI - AMPK induces MUC5B expression via p38 MAPK in NCI-H292 airway epithelial cells. AB - Adenosine monophosphate-activated protein kinase (AMPK) is a well-known serine/threonine kinase that has been implicated in modulation of glucose and fatty acid metabolism. Recent reports have also implicated AMPK in modulation of mucin secretion. In this study, the effects and signaling pathways of AMPK on MUC5B expression were investigated in human NCI-H292 airway epithelial cells. Metformin, as an activator of AMPK, induced MUC5B expression in a dose-dependent manner. Compound C, as an inhibitor of AMPK, inhibited metformin-induced MUC5B expression in a dose-dependent manner. Metformin significantly activated phosphorylation of AMPK; compound C inhibited metformin-activated phosphorylation of AMPK. Without treatment with metformin, there was no difference in MUC5B mRNA expression between Ad-dnAMPK transfected and wild-type adenovirus transfected NCI H292 cells. However, after treatment with metformin, MUC5B mRNA expression was increased in wild-type adenovirus transfected NCI-H292 cells; MUC5B mRNA expression was significantly decreased in Ad-dnAMPK transfected NCI-H292 cells. Metformin activated phosphorylation of p38 mitogen-activated protein kinase (MAPK); compound C inhibited metformin-activated phosphorylation of p38 MAPK. SB203580, as an inhibitor of p38 MAPK, significantly inhibited metformin-induced MUC5B mRNA expression, while U0126, as an inhibitor of ERK1/2 MAPK, had no effect. In addition, knockdown of p38 MAPK by p38 MAPK siRNA significantly blocked metformin-induced MUC5B mRNA expression. In conclusion, results of this study show that AMPK induces MUC5B expression through the p38 MAPK signaling pathway in airway epithelial cells. PMID- 21619870 TI - Adipose tissue-deprived stem cells acquire cementoblast features treated with dental follicle cell conditioned medium containing dentin non-collagenous proteins in vitro. AB - Adipose tissue-derived stem cells (ADSCs), which are easily harvested and show excellent pluripotency potential, have generated considerable interest in regenerative medicine. In this study, the differentiation of ADSCs was assessed after treatment with dental follicle cell conditioned medium (DFCCM) containing dentin non-collagenous proteins (dNCPs). ADSCs exhibited a fibroblast-like morphology and high proliferative capacity. However, after treatment with dNCPs/DFCCM, ADSCs changed from a fibroblast-like to cementoblast-like morphology and significantly lost their proliferative capacity. Alkaline phosphatase activity and in vitro mineralization behaviour of ADSCs were significantly enhanced. Mineralization-related markers including cementum attachment protein, bone sialoprotein, osteocalcin, osteopontin and osteonectin were detected at mRNA or protein levels, whereas dentin sialophosphoprotein and dentin sialoprotein were not detected, implying a cementoblast-like phenotype. These results demonstrate that ADSCs acquired cementoblast features in vitro with dNCPs/DFCCM treatment and could be a potential source of cementogenic cells for periodontal regeneration. PMID- 21619871 TI - OsJAR1 and OsJAR2 are jasmonyl-L-isoleucine synthases involved in wound- and pathogen-induced jasmonic acid signalling. AB - The synthesis of JA-Ile was catalysed by JA-Ile synthase, which is a member of the group I GH3 family of proteins. Here, we showed evidence that OsGH3.5 (OsJAR1) and OsGH3.3 (OsJAR2) are the functional JA-Ile synthases in rice, using recombinant proteins. The expression levels of OsJAR1 and OsJAR2 were induced in response to wounding with the concomitant accumulation of JA-Ile. In contrast, only the expression of OsJAR1 was associated with the accumulation of JA-Ile after blast infection. Our data suggest that these two JA-Ile synthases are differentially involved in the activation of JA signalling in response to wounding and pathogen challenge in rice. PMID- 21619872 TI - Changes in carnitine octanoyltransferase activity induce alteration in fatty acid metabolism. AB - The peroxisomal beta oxidation of very long chain fatty acids (VLCFA) leads to the formation of medium chain acyl-CoAs such as octanoyl-CoA. Today, it seems clear that the exit of shortened fatty acids produced by the peroxisomal beta oxidation requires their conversion into acyl-carnitine and the presence of the carnitine octanoyltransferase (CROT). Here, we describe the consequences of an overexpression and a knock down of the CROT gene in terms of mitochondrial and peroxisomal fatty acids metabolism in a model of hepatic cells. Our experiments showed that an increase in CROT activity induced a decrease in MCFA and VLCFA levels in the cell. These changes are accompanied by an increase in the level of mRNA encoding enzymes of the peroxisomal beta oxidation. In the same time, we did not observe any change in mitochondrial function. Conversely, a decrease in CROT activity had the opposite effect. These results suggest that CROT activity, by controlling the peroxisomal amount of medium chain acyls, may control the peroxisomal oxidative pathway. PMID- 21619873 TI - Akt phosphorylates and regulates the function of Dlx5. AB - Akt, a phosphoinositide-dependent serine/threonine protein kinase, acts as a key regulator in bone formation. Akt can be activated by several osteogenic signaling molecules, but its precise function and downstream targets in bone development are unknown. Dlx5 transcription factor plays important roles during bone development and osteoblast differentiation. Its expression is regulated by several osteogenic signals. In addition, Dlx5 function is also regulated through post-translational modification by several kinases. In this report, we have investigated a potential regulation of Dlx5 function by Akt. Our results indicate that Akt interacts with and phosphorylates Dlx5. In addition, we provide evidences that Akt kinase activity is important for Akt to enhance the protein stability and transcriptional activity of Dlx5. These results suggest that Dlx5 is a novel target of Akt and that the activity of Dlx5 could be modulated by a novel mechanism involving Akt during osteoblast differentiation. PMID- 21619874 TI - A novel C2 transferrin variant interfering with the analysis of carbohydrate deficient transferrin. AB - BACKGROUND: Carbohydrate-deficient transferrin (CDT) is used as a marker for chronic alcohol abuse. The presence of genetic transferrin variants might affect an individual's iron status and can interfere with CDT analysis. We report on the identification of a patient carrying a novel transferrin variant. We describe the performance of the various CDT methods in its detection and the associated iron status. METHODS: DNA of the coding region of transferrin was sequenced and CDT levels were analysed using four different methods: high pressure liquid chromatography (HPLC), capillary zone electrophoresis (CZE), immunochemistry and iso-electric focussing (IEF). RESULTS: A novel transferrin variant, T139M, was found as a heterozygous genotype in the index patient and all of his four living direct family members (c.416C>p.Thr139Met). CDT analysis of the variant by HPLC and CZE was compromised as a result of the coelution of the different isoforms. CDT levels could be quantified by immunochemistry. Similar results were obtained using IEF analysis. The presence of the C2 transferrin variant did not affect iron status in any of the investigated patients. CONCLUSIONS: Transferrin T139M, present as a heterozygous genotype, interferes with CDT analysis by HPLC and CZE but not by immunochemistry. Physiologically, it appears to be functionally normal. PMID- 21619875 TI - Care during freeze-drying of bovine pericardium tissue to be used as a biomaterial: a comparative study. AB - Bovine pericardium (BP) tissue is widely used in the manufacture of bioprosthetics. The effects of freeze-drying on the BP tissue have been studied by some researchers in order to decrease their cytotoxicity due to preservation in formaldehyde solution, and to increase the lifetime of the product in storage. This study was undertaken in order to study the effect of freeze-drying in the structure of BP. To perform this study BP samples were freeze-dried in two different types of freeze-dryers available in our laboratory: a laboratory freeze dryer, in which it was not possible to control parameters and a pilot freeze dryer, wherein all parameters during freezing and drying were controlled. After freeze-drying processes, samples were analyzed by SEM, Raman spectroscopy, tensile strength, water uptake tests and TEM. In summary, it has been demonstrated that damages occur in collagen fibers by the loss of bulk water of collagen structure implicating in a drastic decreasing of BP mechanical properties due to its structural alterations. Moreover, it was proven that the collagen fibrils suffered breakage at some points, which can be attributed to the uncontrolled parameters during drying. PMID- 21619876 TI - The R(h)oads to Stat3: Stat3 activation by the Rho GTPases. AB - The signal transducer and activator of transcription-3 (Stat3) is a member of the STAT family of cytoplasmic transcription factors. Overactivation of Stat3 is detected with high frequency in human cancer and is considered a molecular abnormality that supports the tumor phenotype. Despite concerted investigative efforts, the molecular mechanisms leading to the aberrant Stat3 activation and Stat3-mediated transformation and tumorigenesis are still not clearly defined. Recent evidence reveals a crosstalk close relationship between Stat3 signaling and members of the Rho family of small GTPases, including Rac1, Cdc42 and RhoA. Specifically, Rac1, acting in a complex with the MgcRacGAP (male germ cell RacGAP), promotes tyrosine phosphorylation of Stat3 by the IL6-receptor family/Jak kinase complex, as well as its translocation to the nucleus. Studies have further revealed that the mutational activation of Rac1 and Cdc42 results in Stat3 activation, which occurs in part through the upregulation of IL6 family cytokines that in turn stimulates Stat3 through the Jak kinases. Interestingly, evidence also shows that the engagement of cadherins, cell to cell adhesion molecules, specifically induces a striking increase in Rac1 and Cdc42 protein levels and activity, which in turn results in Stat3 activation. In this review we integrate recent findings clarifying the role of the Rho family GTPases in Stat3 activation in the context of malignant progression. PMID- 21619877 TI - The STAT3 beacon: IL-6 recurrently activates STAT 3 from endosomal structures. AB - Endocytic trafficking plays an important role in signal transduction. Signal transducer and activator of transcription 3 (STAT3) and mitogen-activate protein kinase (MAPK) have both been localized to endosomal structures and are dependent upon endocytosis for downstream function. While the dependence of MAPK signaling upon endosomes has been well characterized, the involvement of endosomes in regulating STAT3 signaling has not been defined. Consequently, this study evaluated the role of endosomes in the initiation, modulation, amplification and persistence of interleukin-6(IL-6)-induced STAT3 signal transduction and transcription, and utilized IL-6-induced MAPK signaling as a comparator. Using pharmacologic treatment and temperature control of endocytic trafficking, pulse chase treatments and in vitro kinase assays, STAT3 was found to interact with endosomes in a markedly different fashion than MAPK. STAT3 was activated by direct interaction with internal structures upstream of the late endosome following IL-6 exposure and persistent STAT3 signaling depended upon recurrent activation from endocytic structures. Further, STAT3 subcellular localization was not dependent upon endocytic trafficking. Instead, STAT3 transiently interacted with endosomes and relocated to the nucleus by an endosome-independent mechanism. Finally, endocytic trafficking played a central role in regulating STAT3 serine 727 phosphorylation through crosstalk with the MAPK signaling system. Together, these data reveal endosomes as central to the genesis, course and outcome of STAT3 signal transduction and transcription. PMID- 21619878 TI - Kinematic analyses reveal impaired locomotion following injury of the motor cortex in mice. AB - Brain injury in the motor cortex can result in deleterious functional deficits of skilled and fine motor functions. However, in contrast to humans, the destruction of cortex and its descending fibers has been thought not to cause remarkable deficits in simple locomotion in quadropedal animals. In the present study, we aimed to investigate in detail how lesion of the sensorimotor cortex affected locomotion ability in mice using the KinemaTracer system, a novel video-based kinematic analyzer. We found that traumatic injury to the left sensorimotor cortex induced several apparent deficits in the movement of contralesional right limbs during treadmill locomotion. The step length of right limbs decreased, and the speed in the forward direction was abrogated in the swing phase. The coordinates and angle of each joint were also changed after the injury. Some of the abnormal values in these parameters gradually recovered near the control level. The number of cFos-expressing neurons following locomotion significantly decreased in the right side of the spinal cord in injured mice, suggesting a role for cortex and descending fibers in locomotion. In contrast, interlimb coordination did not change remarkably even after the injury, supporting the notion that the basic locomotor pattern was determined by intraspinal neural circuits. These results indicate that the motor cortex and its descending fibers regulate several aspects of fine limb movement during locomotion. Our findings provide practical parameters to assess motor deficits and recovery following cortical injury in mice. PMID- 21619879 TI - Adaptation to moderate hypoxia protects cortical neurons against ischemia reperfusion injury and excitotoxicity independently of HIF-1alpha. AB - Continuous exposure of cultured cortical neurons to moderate hypoxia (1% O(2)) elevates cellular accumulation of hypoxia-inducible factor-1alpha (HIF-1alpha) and improves basal survival of cultured cortical neurons. We examined the effects of adaptation to moderate hypoxia on the vulnerability of cultured neurons to the acute injury of simulated ischemia-reperfusion. Cortical neurons cultured continuously in 1% O(2) were markedly protected against simulated ischemia reperfusion, with protection persisting through 72h after ischemia. Neurons from 1% O(2) conditions were also highly resistant to glutamate-induced NMDA receptor dependent excitotoxic injury, despite expression of NMDA receptors at levels not significantly changed from controls. Inhibition of prolyl hydroxylase, mimicking cellular signaling effects of hypoxia including HIF-1alpha stabilization, also protected neurons against simulated ischemia-reperfusion injury. Nevertheless, genetic deletion of HIF-1alpha expression did not diminish the protection of neurons adapted to 1% O(2) from excitotoxicity or ischemia-reperfusion injury, nor did it prevent the protective effect of prolyl hydroxylase inhibition. We conclude that chronic exposure to moderate hypoxia, through HIF-1alpha independent mechanisms, produces strong protective effects against excitotoxic and ischemia-reperfusion related injury. PMID- 21619880 TI - Differentiation of Entamoeba histolytica: a possible role for enolase. AB - The study of the encystation process of Entamoeba histolytica has been hampered by the lack of experimental means of inducing mature cysts in vitro. Previously we have found that cytoplasmic vesicles similar to the encystation vesicles of Entamoeba invadens are present in E. histolytica trophozoites only in amebas recovered from experimental amebic liver abscesses. Here we report that a monoclonal antibody (B4F2) that recognizes the cyst wall of E. invadens also identifies a 48 kDa protein in vesicles of E. histolytica trophozoites recovered from hepatic lesions. This protein is less expressed in trophozoites continuously cultured in axenical conditions. As previously reported for E. invadens, the B4F2 specific antigen was identified as enolase in liver-recovered E. histolytica, by two-dimensional electrophoresis, Western blot and mass spectrometry. In addition, the E. histolytica enolase mRNA was detected by RT PCR. The antigen was localized by immunoelectron microscopy in cytoplasmic vesicles of liver-recovered amebas. The B4F2 antibody also recognized the wall of mature E. histolytica cysts obtained from human samples. These results suggest that the enolase-containing vesicles are produced by E. histolytica amebas, when placed in the unfavorable liver environment that could be interpreted as an attempt to initiate the encystation process. PMID- 21619881 TI - Taenia solium: a two-dimensional Western blotting method combined with the use of an EST-library for the identification of immunogenic proteins recognized by sera from neurocysticercosis patients. AB - Commercial antigens used to diagnose human neurocysticercosis (NCC) are obtained from either a soluble parasite extract or a parasite-derived glycoprotein fraction. The aim of the present study was to identify antigenic proteins as potential diagnostic candidates in this context. Soluble immunogenic proteins from Taenia solium cysticerci were identified by two-dimensional electrophoresis Western blotting using human sera from Nicaragua confirmed to be positive for NCC by computer tomography. Six antigenic proteins were identified and sequenced by liquid chromatography-mass spectrometry. Among these immunogenic proteins, a novel sequence was found and named Tsol-p27. To determine the antigenicity of Tsol-p27, the previously reported antigen TsolHSP36 and the new Tsol-p27 were expressed as recombinant proteins and evaluated serologically. Immunoblotting demonstrated that Tsol-p27 was recognized by sera from 13 NCC-positive humans, whereas TsolHSP36 was identified by only two of those 13 positive sera. None of the antigens were recognized by negative control sera. Despite the limited number of serum samples evaluated in this study, the results indicate that Tsol-p27 might be a suitable candidate for diagnosis of human NCC. PMID- 21619883 TI - Real time-PCR assay estimating the naive T-cell pool in whole blood and dried blood spot samples: pilot study in young adults. AB - Because of their central role orchestrating the immune response, the decrease in repertoire number and diversity of naive T-cells is a significant feature of immnosenescence. Reflecting the effective naive T-cell pool, quantifying the sj TREC ratio (number of signal joint T-cell receptor excision circles/10(5) T cells) in blood samples suffers however from constraints. The most limiting one is the absolute requirement of the flow cytometry analysis of peripheral blood samples for the T-cell numeration. In order to make this ratio more accessible for clinical and epidemiological studies addressing how changes in responsiveness of the immune system lead to an increased susceptibility to various diseases and poorer response to vaccination, we have developed a rapid and simple method for the quantification of the sj-TREC ratio in whole blood and in dried blood spot (DBS) samples. This novel method is a QPCR analysis using fluorescently labelled sequence-specific probes both for quantifying sj-TREC and T-cell count and therefore eliminating the absolute necessity of the flow cytometer analysis. In this pilot study, we have compared the sj-TREC ratio we obtained with this novel method in whole blood and in DBS samples of 10 healthy volunteers with those obtained with the technique of reference and found that they are comparable. PMID- 21619882 TI - Negative emotionality, depressive symptoms and cortisol diurnal rhythms: analysis of a community sample of middle-aged males. AB - Prior research suggests that individuals with particular personality traits, like negative emotionality, are at greater risk for adverse health outcomes. Despite bivariate associations between negative emotionality, depressive symptoms and the hypothalamic pituitary adrenal axis (HPA axis), few studies have sought to understand the biological pathways through which negative emotionality, depressive symptomatology and cortisol-one of the primary hormonal products of the HPA axis--are associated. The present study explored whether negative emotionality influenced cortisol dysregulation through current depressive symptomatology and whether negative emotionality served as a moderator of the relationship between depressive symptoms and cortisol. In the community-based Vietnam Era Twin Study of Aging, 783 male twins completed two days of cortisol saliva sampling in their natural environments. Three measures of cortisol were analyzed: waking levels, the cortisol awakening response, and the peak to bed slope. Depressive symptoms significantly mediated the associations between negative emotionality and the peak to bed slope. A 2-way interaction between depressive symptoms and negative emotionality was significant for the peak to bed slope and for waking levels of cortisol. Exploration of the interactions illustrated that depressive symptoms only affected cortisol slopes at average or high levels of negative emotionality and only affected waking levels at low levels of negative emotionality. Negative emotionality and depressive symptoms were not related to the cortisol awakening response. This is the first study to find indirect associations between negative emotionality and peak to bed cortisol slopes through depressive symptoms. These findings illustrate the complex interplay between personality characteristics, depressive symptoms and different indices of the cortisol diurnal rhythm. PMID- 21619884 TI - A proteasome assembly defect in rpn3 mutants is associated with Rpn11 instability and increased sensitivity to stress. AB - Rpn11 is a proteasome-associated deubiquitinating enzyme that is essential for viability. Recent genetic studies showed that Rpn11 is functionally linked to Rpn10, a major multiubiquitin chain binding receptor in the proteasome. Mutations in Rpn11 and Rpn10 can reduce the level and/or stability of proteasomes, indicating that both proteins influence its structural integrity. To characterize the properties of Rpn11, we examined its interactions with other subunits in the 19S regulatory particle and detected strong binding to Rpn3. Two previously described rpn3 mutants are sensitive to protein translation inhibitors and an amino acid analog. These mutants also display a mitochondrial defect. The abundance of intact proteasomes was significantly reduced in rpn3 mutants, as revealed by strongly reduced binding between 20S catalytic with 19S regulatory particles. Proteasome interaction with the shuttle factor Rad23 was similarly reduced. Consequently, higher levels of multiUb proteins were associated with Rad23, and proteolytic substrates were stabilized. The availability of Rpn11 is important for maintaining adequate levels of intact proteasomes, as its depletion caused growth and proteolytic defects in rpn3. These studies suggest that Rpn11 is stabilized following its incorporation into proteasomes. The instability of Rpn11 and the defects of rpn3 mutants are apparently caused by a failure to recruit Rpn11 into mature proteasomes. PMID- 21619885 TI - Copper(II)-induced secondary structure changes and reduced folding stability of the prion protein. AB - The cellular isoform of the prion protein PrP(C) is a Cu(2)(+)-binding cell surface glycoprotein that, when misfolded, is responsible for a range of transmissible spongiform encephalopathies. As changes in PrP(C) conformation are intimately linked with disease pathogenesis, the effect of Cu(2+) ions on the structure and stability of the protein has been investigated. Urea unfolding studies indicate that Cu(2+) ions destabilise the native fold of PrP(C). The midpoint of the unfolding transition is reduced by 0.73 +/- 0.07 M urea in the presence of 1 mol equiv of Cu(2+). This equates to an appreciable difference in free energy of unfolding (2.02 +/- 0.05 kJ mol(-1) at the midpoint of unfolding). We relate Cu(2)(+)-induced changes in secondary structure for full-length PrP(23 231) to smaller Cu(2)(+) binding fragments. In particular, Cu(2+)-induced structural changes can directly be attributed to Cu(2+) binding to the octarepeat region of PrP(C). Furthermore, a beta-sheet-like transition that is observed when Cu ions are bound to the amyloidogenic fragment of PrP (residues 90-126) is due only to local Cu(2+) coordination to the individual binding sites centred at His95 and His110. Cu(2+) binding does not directly generate a beta-sheet conformation within PrP(C); however, Cu(2+) ions do destabilise the native fold of PrP(C) and may make the transition to a misfolded state more favourable. PMID- 21619886 TI - An SIS patch model with variable transmission coefficients. AB - In this paper, an SIS patch model with non-constant transmission coefficients is formulated to investigate the effect of media coverage and human movement on the spread of infectious diseases among patches. The basic reproduction number R(0) is determined. It is shown that the disease-free equilibrium is globally asymptotically stable if R(0)<=1, and the disease is uniformly persistent and there exists at least one endemic equilibrium if R(0)>1. In particular, when the disease is non-fatal and the travel rates of susceptible and infectious individuals in each patch are the same, the endemic equilibrium is unique and is globally asymptotically stable as R(0)>1. Numerical calculations are performed to illustrate some results for the case with two patches. PMID- 21619887 TI - The selective phosphodiesterase 9 (PDE9) inhibitor PF-04447943 (6-[(3S,4S)-4 methyl-1-(pyrimidin-2-ylmethyl)pyrrolidin-3-yl]-1-(tetrahydro-2H-pyran-4-yl)-1,5 dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one) enhances synaptic plasticity and cognitive function in rodents. AB - Inhibition of phosphodiesterase 9 (PDE9) has been reported to enhance rodent cognitive function and may represent a potential novel approach to improving cognitive dysfunction in Alzheimer's disease. PF-04447943, (6-[(3S,4S)-4-methyl-1 (pyrimidin-2-ylmethyl)pyrrolidin-3-yl]-1-(tetrahydro-2H-pyran-4-yl)-1,5-dihydro 4H-pyrazolo[3,4-d]pyrimidin-4-one), a recently described PDE9 inhibitor, was found to have high affinity (Ki of 2.8, 4.5 and 18 nM) for human, rhesus and rat recombinant PDE9 respectively and high selectivity for PDE9 versus PDEs1-8 and 10 11. PF-04447943 significantly increased neurite outgrowth and synapse formation (as indicated by increased synapsin 1 expression) in cultured hippocampal neurons at low (30-100 nM) but not high (300-1000 nM) concentrations. PF-04447943 significantly facilitated hippocampal slice LTP evoked by a weak tetanic stimulus at a concentration of 100 nM but failed to affect response to the weak tetanus at either 30 or 300 nM, or the LTP produced by a theta burst stimulus. Systemic administration of PF-04447943 (1-30 mg/kg p.o.) dose-dependently increased cGMP in the cerebrospinal fluid 30 min after administration indicating target engagement in the CNS of rats. PF-04447943 (1-3 mg/kg p.o.) significantly improved cognitive performance in three rodent cognition assays (mouse Y maze spatial recognition memory model of natural forgetting, mouse social recognition memory model of natural forgetting and rat novel object recognition with a scopolamine deficit). When administered at a dose of 3 mg/kg p.o., which improved performance in novel object recognition, PF-04447943 significantly increased phosphorylated but not total GluR1 expression in rat hippocampal membranes. Collectively these data indicate that PF-04447943 is a potent, selective brain penetrant PDE9 inhibitor that increased indicators of hippocampal synaptic plasticity and improved cognitive function in a variety of cognition models in both rats and mice. Results with PF-04447943 are consistent with previously published findings using a structurally diverse PDE9 inhibitor, BAY73-6199, and further support the suggestion that PDE9 inhibition may represent a novel approach to the palliative remediation of cognitive dysfunction. PMID- 21619888 TI - Raclopride lessens the ability of clozapine to suppress alcohol drinking in Syrian golden hamsters. AB - Emerging evidence suggests that the atypical antipsychotic clozapine decreases alcohol consumption in patients with schizophrenia, while typical antipsychotics, all of which are potent dopamine (DA) D2 receptor antagonists, do not. We have proposed that clozapine, through its weak DA D2 receptor blocking action, coupled with its ability to potentiate noradrenergic and serotonergic activity, may ameliorate a dysfunction in the mesocorticolimbic DA reward circuitry that underlies alcohol use disorder in patients with schizophrenia. In prior studies, we have demonstrated that clozapine also decreases alcohol drinking in the Syrian golden hamster, but haloperidol does not. The purposes of the current study were: (1) to further assess the effect of clozapine (2 or 4 mg/kg/day, s.c.) on alcohol consumption in hamsters, using a continuous access, 2-bottle choice paradigm; and (2) to examine whether clozapine's effect on alcohol drinking is affected by increasing its DA D2 blockade through adjunctive use of the potent DA D2 receptor antagonist raclopride (2, 4, or 6 mg/kg/day, s.c.). The major findings were: (1) clozapine suppressed both initiation and maintenance of alcohol drinking in hamsters; and (2) these effects of clozapine were lessened when raclopride was given adjunctively with clozapine. These data suggest that clozapine may limit alcohol drinking in the golden hamster (and possibly in patients with schizophrenia) in part because of its weak blockade of the DA D2 receptor. PMID- 21619889 TI - mGluR2/3 agonist LY379268, by enhancing the production of GDNF, induces a time related phosphorylation of RET receptor and intracellular signaling Erk1/2 in mouse striatum. AB - In the present study we aimed to verify if the enhancement of glial cell line derived neurotrophic factor (GDNF) production in mouse striatum following treatment with LY379268 may also induce in the nigrostriatal system a time related activation of RET receptor and its specific intracellular signaling. For this purpose, we have investigated the effects of LY379268 treatment on RET phosphorylation at the Tyr1062 and on downstream signaling Erk1/2, Akt and PLCgamma1 pathway activation. The results showed that treatment with LY379268 (3 mg/kg) induces a significant increase of GDNF levels and time-related RET and Erk1/2 phosphorylation in the striatum. These increases were detected at 24 h and 48 h following LY379268 treatment. No changes were observed in the Akt and PLCgamma1 phosphorylation levels. Similar results for p-Erk1/2 were observed in the substantia nigra. A complete block of LY379268 effect on striatal RET and p Erk1/2 phosphorylation was observed in mice intrastriatal injected with anti-GDNF antibodies, suggesting a correlation between GDNF upregulation and RET activation. Overall, with present data we have shown that activation of mGluR2/3 receptors by LY379268 may be particularly promising for nigrostriatal dopaminergic system protection by enhancing striatal levels of GDNF/RET trophic system activity. PMID- 21619890 TI - Activation of 5-HT6 receptors inhibits corticostriatal glutamatergic transmission. AB - We investigated the effect of 5-HT6 receptor subtype activation on glutamatergic transmission by means of whole-cell patch-clamp electrophysiological recordings from medium spiny neurons of the striatum and layer V pyramidal neurons of the prefrontal cortex. To this aim, we took advantage of a novel ligand, ST1936, showing nM affinity and agonist activity at the 5-HT6 receptor subtype. Our data show that 5-HT6 receptor activation by ST1936 reduces the frequency of spontaneous excitatory postsynaptic currents, with an IC50 of 1.3 MUM. Moreover, 5-HT6 receptor activation also reduced the amplitude of spontaneous excitatory postsynaptic currents recorded from medium spiny neurons, suggesting a mechanism of action involving postsynaptic 5-HT6 receptors, as further confirmed by the paired-pulse analysis on evoked excitatory postsynaptic currents and by recordings of miniature glutamatergic events. The inhibitory effect of ST1936 on glutamatergic transmission was prevented by the selective 5-HT6 receptor antagonist SB258585 and mimicked by a different agonist, WAY-181187. Conversely, in the cortex ST1936 reduced the frequency, but not the amplitude, of spontaneous excitatory postsynaptic currents suggesting a presynaptic or indirect effect of the 5-HT6 receptor. PMID- 21619891 TI - Modulation of gap-junctional intercellular communication by a series of cyanobacterial samples from nature and laboratory cultures. AB - Cyanobacterial extracts have been recently shown to alter two in vitro biomarkers of tumor promotion, namely to cause inhibition of gap-junctional intercellular communication (GJIC) and activation of mitogen-activated protein kinases (Blaha et al., 2010a). In the present study, we investigated GJIC-inhibitory potencies of 10 laboratory strains representing common water bloom-forming cyanobacteria (Anabaena, Aphanizomenon, Cylindrospermopsis, Microcystis and Planktothrix) and six natural water bloom samples (dominated by Aphanizomenon sp. or Microcystis). The most pronounced inhibitions of GJIC in a model rat liver epithelial cell line WB-F344 were caused by methanolic extracts of Anabaena flos-aquae UTEX 1444, Aphanizomenon flos-aquae SAG 31.87, Aphanizomenon gracile RCX 06, Microcystis aeruginosa PCC 7806, Cylindrospermopsis raciborskii SAG 1.97, Planktothrix agardhii CCALA 159 and SAG 32.79, whereas weaker effects were induced by Aphanizomenon klebahnii CCALA 009 and no inhibition was induced by extracts of Aph. flos-aquae PCC 7905 and Aph. gracile SAG 31.79. Exudates of the laboratory cultured strains concentrated by solid phase extraction also induced species specific inhibitory effects, but they did not necessarily correlate with the inhibitory potencies of extracts from the corresponding species. Interestingly, the GJIC-inhibitory effects may not be restricted to cyanobacteria, since exudates of two green alga species also affected GJIC, although their extracts caused no effects. The extracts from different natural water blooms inhibited GJIC with different potencies without apparent relation to bloom-species composition. Since the observed effects on GJIC did not correlate with the content of cyanotoxins microcystins and cylindrospermopsin in the tested samples, they were most likely induced by unknown compound(s). Our results indicate that putative tumor promoting compound(s) could be associated with different species of bloom-forming cyanobacteria, but their production is probably species- and strain-specific. PMID- 21619892 TI - Astaxanthin limits fish oil-related oxidative insult in the anterior forebrain of Wistar rats: putative anxiolytic effects? AB - The habitual consumption of marine fish is largely associated to human mental health. Fish oil is particularly rich in n-3 polyunsaturated fatty acids that are known to play a role in several neuronal and cognitive functions. In parallel, the orange-pinkish carotenoid astaxanthin (ASTA) is found in salmon and displays important antioxidant and anti-inflammatory properties. Many neuronal dysfunctions and anomalous psychotic behavior (such as anxiety, depression, etc.) have been strongly related to the higher sensitivity of cathecolaminergic brain regions to oxidative stress. Thus, the aim of this work was to study the combined effect of ASTA and fish oil on the redox status in plasma and in the monoaminergic-rich anterior forebrain region of Wistar rats with possible correlations with the anxiolytic behavior. Upon fish oil supplementation, the downregulation of superoxide dismutase and catalase activities combined to increased "free" iron content resulted in higher levels of lipid and protein oxidation in the anterior forebrain of animals. Such harmful oxidative modifications were hindered by concomitant supplementation with ASTA despite ASTA related antioxidant protection was mainly observed in plasma. Although it is clear that ASTA properly crosses the brain-blood barrier, our data also address a possible indirect role of ASTA in restoring basal oxidative conditions in anterior forebrain of animals: by improving GSH-based antioxidant capacity of plasma. Preliminary anxiolytic tests performed in the elevated plus maze are in alignment with our biochemical observations. PMID- 21619893 TI - Nanoparticle-based bio-barcode assay for the detection of bluetongue virus. AB - The ultrasensitive bio-barcode amplification assay (BCA) technique was developed for the specific detection of the outer-core protein VP7 of bluetongue virus (BTV). The target antigen VP7 was first captured by gold nanoparticles (GNPs) coated with polyclonal antibodies. Magnetic microparticles (MMP) coated with VP7 monoclonal antibody were then added to form a sandwich immuno-complex. After the immuno-complex was formed, signal DNA annealed to DNA strands covalently bound to the GNPs were released by heating and characterized by PCR and real-time fluorescence PCR. A detection limit of 0.1fg/ml was measured for purified VP7, seven orders of magnitude more sensitive than that of conventional antigen capture ELISA. The BCA demonstrated the same enhanced sensitivity for detecting BTV in serum samples from sheep. In the following work it is demonstrated that this assay is a highly sensitive method for the detection of BTV proteins that could be adapted to measure other proteins. PMID- 21619894 TI - Rapid screening and confirmatory methods for biochemical diagnosis of human prion disease. AB - Transmissible spongiform encephalopathies (TSEs) are characterised by accumulation of an abnormal isoform of prion protein (PrP(sc)), mainly in the brain but also in various peripheral tissues. Home-made assays consisting of non standardised protocols are used currently for laboratory diagnosis of human TSE. The purpose of the present study was to test the ability of two commercial assays, TeSeETM CJD ELISA and TeSeETM Western blot, to detect PrPsc in cerebral and lymphoid tissues of TSE patients. Both tests detected a PrPsc-significant signal in the brains of 54 affected patients and not in 51 controls, yielding 100% specificity and 100% sensitivity. Furthermore, three post-mortem spleens and two pre-mortem tonsils from three patients with variant Creutzfeldt-Jakob disease (vCJD) were detected correctly. The expected PrPsc molecular patterns were found in TSE patient brain tissue and in the tonsils and spleens of the three vCJD patients. In conclusion, these rapid and robust in vitro tools were suitable for routine human TSE diagnosis and characterisation. CJD could also be diagnosed during the patient's lifetime by detection of PrPsc in the tonsil. A diagnostic strategy associating TeSeETM CJD ELISA screening to biochemical confirmation by TeSeETM Western blot is proposed. PMID- 21619895 TI - Detection of shrimp Taura syndrome virus by loop-mediated isothermal amplification using a designed portable multi-channel turbidimeter. AB - In this study, a portable turbidimetric end-point detection method was devised and tested for the detection of Taura syndrome virus (TSV) using spectroscopic measurement of a loop-mediated isothermal amplification (LAMP) by-product: magnesium pyrophosphate (Mg(2)P(2)O(7)). The device incorporated a heating block that maintained an optimal temperature of 63 degrees C for the duration of the RT LAMP reaction. Turbidity of the RT-LAMP by-product was measured when light from a light-emitting diode (LED) passed through the tube to reach a light dependent resistance (LDR) detector. Results revealed that turbidity measurement of the RT LAMP reactions using this device provided the same detection sensitivity as the agarose gel electrophoresis detection of RT-LAMP and nested RT-PCR (IQ2000TM) products. Cross reactions with other shrimp viruses were not found, indicating that the RT-LAMP-turbidity measurement was highly specific to TSV. The combination of 10 min for rapid RNA preparation with 30 min for RT-LAMP amplification followed by turbidity measurement resulted in a total assay time of less than 1h compared to 4-8h for the nested RT-PCR method. RT-LAMP plus turbidity measurement constitutes a platform for the development of more rapid and user-friendly detection of TSV in the field. PMID- 21619896 TI - Cell-based analysis of Chikungunya virus membrane fusion using baculovirus expression vectors. AB - Chikungunya virus infection has emerged in many countries over the past decade. There are no effective drugs for controlling the disease. To develop cell-based system for screening anti-virus drugs, a bi-cistronic baculovirus expression system was utilized to co-express viral structural proteins C (capsid), E2 and E1 and the enhanced green fluorescence protein (EGFP) in Spodoptera frugiperda insect cells (Sf21). The EGFP-positive Sf21 cells fused with each other and with uninfected cells to form a syncytium, allowing characterization of cholesterol and low pH requirements for syncytium formation. Western blot analysis showed three structural proteins were expressed in baculovirus infected cells. The structural proteins of Chikungunya virus that is required for cell fusion was determined with various recombinant baculoviruses bearing different lengths of the viral structural protein genes. Protein E1 was required for cell fusion and indicating that Chikungunya viral membrane fusion was a class II membrane fusion. It was also demonstrated that the heterologous expression of alphavirus monomeric E1 can induce insect cell fusions. Furthermore, this cell-based system provides a model for studying class II viral membrane fusion. PMID- 21619897 TI - Acridine derivatives as anti-BVDV agents. AB - Twenty-six 9-aminoacridine derivatives were evaluated in cell-based assays for cytotoxicity and antiviral activity against a panel of 10 RNA and DNA viruses. While seven compounds (9, 10, 14, 19, 21, 22, 24) did not affect any virus and two (6, 11) were moderately active against CVB-5 or Reo-1, 17 compounds exhibited a marked specific activity against BVDV, prototype of pestiviruses which are responsible for severe diseases of livestock. Most anti-BVDV agents showed EC(50) values in the range 0.1-8 MUM, thus comparing favorably with the reference drugs ribavirine and NM 108. Some compounds, particularly those bearing a quinolizidinylalkyl side chain, displayed pronounced cytotoxicity. Further studies are warranted in order to achieve still better anti-BVDV agents, and to explore the potential antiproliferative activity of this kind of compounds. PMID- 21619898 TI - Effects of nevirapine and efavirenz on human adipocyte differentiation, gene expression, and release of adipokines and cytokines. AB - The non-nucleoside reverse transcriptase inhibitors (NNRTIs) nevirapine and efavirenz are drugs of choice for initial antiretroviral treatment for HIV-1 infection. Although NNRTIs have not traditionally been associated with the appearance of adipose alterations, recent data suggest that efavirenz may contribute to adipose tissue alterations in antiretroviral-treated patients, consistent with its ability to impair differentiation of adipocytes in cell cultures. No such effects have been reported for nevirapine, the other most commonly used NNRTI. In this study, we determined the effects of nevirapine on differentiation, gene expression and release of regulatory proteins (adipokines and cytokines) in differentiating human adipocytes, and compared them with those of efavirenz. Efavirenz caused a dose-dependent repression of adipocyte differentiation that was associated with down-regulation of the master adipogenesis regulator genes SREBP-1, PPARgamma and C/EBPalpha, and their target genes encoding lipoprotein lipase, leptin and adiponectin, which are key proteins in adipocyte function. In contrast, nevirapine does not affect adipogenesis and causes a modest but significant coordinate increase in the expression of SREBP-1, PPARgamma and C/EBPalpha and their target genes only at a concentration of 20 MUM. Whereas efavirenz caused a significant increase in the release of pro inflammatory cytokines (interleukin [IL]-8, IL-6, monocyte chemoattractant protein-1), plasminogen activator inhibitor type-1 and hepatocyte growth factor (HGF), nevirapine either had no effect on these factors or decreased their release (IL-6 and HGF). Nevirapine significantly increased adiponectin release, whereas efavirenz strongly repressed it. Moreover, nevirapine inhibited preadipocyte endogenous reverse transcriptase activity, whereas efavirenz did not alter it. It is concluded that, in contrast with the profound anti-adipogenic and pro-inflammatory response elicited by efavirenz, nevirapine does not impair adipogenesis. PMID- 21619899 TI - Electrical responses to chemosensory stimulation recorded from the vomeronasal duct and the respiratory epithelium in humans. AB - The physiological significance of the human vomeronasal duct (VND) is still unclear. The aim of the present study was to investigate the question whether mucosal responses obtained from the VND are different from those obtained from the respiratory epithelium. There were 15 healthy subjects (8 male, 7 female; age range 19-45 years; 14 normosmic subjects, 1 anosmic subject). All subjects participated in two sessions whereby the first session was used to acquaint them with the experimental conditions. For chemical stimulation, an olfactometer was used which delivered chemical stimulants without altering mechanical or thermal conditions at the stimulated nasal mucosa. For stimulation we used substances previously reported to produce vomeronasal activation ("estra"=estra-1,3,5(10),16 tetraen-3ol and "andro"=androsta-4,16-dien-3-on); in addition, gaseous CO(2) was used as a non-odorous, relatively specific stimulant of the trigeminal nerve. Placement of electrodes either in the VND or on the respiratory epithelium was performed under endoscopical guidance. Subjects rated the overall intensity of the stimuli, the strength of trigeminally mediated sensations, and the hedonic tone of the stimulants. Responses could not be recorded from all subjects. For the remaining 7 subjects, intensity was strongest for CO(2) stimuli (p<0.001), whereas no significant difference was observed between "andro" and "estra" (p=0.33). All three stimulants produced responses at the respiratory epithelium with largest responses obtained after stimulation with CO(2). Similar findings were made for recordings inside the VND. Due to the small sample size sexual dimorphisms could not be addressed. In summary, these results seem to indicate that the presently used stimulants produce similar responses at the respiratory epithelium and in the VND which argues against a specific responsiveness of the VND epithelium to chemosensory stimuli although it has to be kept in mind that the effective sample size was small. PMID- 21619900 TI - Does intolerance of uncertainty predict anticipatory startle responses to uncertain threat? AB - Intolerance of uncertainty (IU) has been proposed to be an important maintaining factor in several anxiety disorders, including generalized anxiety disorder, obsessive-compulsive disorder, and social phobia. While IU has been shown to predict subjective ratings and decision-making during uncertain/ambiguous situations, few studies have examined whether IU also predicts emotional responding to uncertain threat. The present study examined whether IU predicted aversive responding (startle and subjective ratings) during the anticipation of temporally uncertain shocks. Sixty-nine participants completed three experimental conditions during which they received: no shocks, temporally certain/predictable shocks, and temporally uncertain shocks. Results indicated that IU was negatively associated with startle during the uncertain threat condition in that those with higher IU had a smaller startle response. IU was also only related to startle during the uncertain (and not the certain/predictable) threat condition, suggesting that it was not predictive of general aversive responding, but specific to responses to uncertain aversiveness. Perceived control over anxiety related events mediated the relation between IU and startle to uncertain threat, such that high IU led to lowered perceived control, which in turn led to a smaller startle response. We discuss several potential explanations for these findings, including the inhibitory qualities of IU. Overall, our results suggest that IU is associated with attenuated aversive responding to uncertain threat. PMID- 21619901 TI - Interactions in a host plant-virus-vector-parasitoid system: modelling the consequences for virus transmission and disease dynamics. AB - A full understanding of plant virus epidemiology requires studies at different scales of integration: from within-plant cell processes to vector population dynamics, behaviour and broader ecological interactions. Vectors respond to cues derived from plants (both healthy and virus-infected), from natural enemies and from other environmental influences, and these directly affect the temporal and spatial patterns of disease development. The key element in linking these scales is the transmission process and the determining factors involved. We use a mathematical model to show how the presence of natural enemies, by increasing virus transmission, can increase the rate of virus disease development while at the same time reducing vector population size, supporting recent empirical evidence obtained in microcosm studies. The implication of this work is that biological control of arthropod pests, which are also virus vectors, using parasitoid wasps, may have unanticipated and negative effects in terms of increased incidence of virus disease. PMID- 21619902 TI - Isolation and partial characterization of Brazilian samples of feline immunodeficiency virus. AB - Feline immunodeficiency virus (FIV) causes a slow progressive degeneration of the immune system which eventually leads to a disease comparable to acquired immune deficiency syndrome (AIDS) in humans. FIV has extensive sequence variation, a typical feature of lentiviruses. Sequence analysis showed that diversity was not evenly distributed throughout the genome, but was greatest in the envelope gene, env. The virus enters host cells via a sequential interaction, initiated by the envelope glycoprotein (env) binding the primary receptor molecule CD134 and followed by a subsequent interaction with chemokine co-receptor CXCR4. The purpose of this study was to isolate and characterize isolates of FIV from an open shelter in Sao Paulo, Brazil. The separated PBMC from 11 positive cats were co-cultured with MYA-1 cells. Full-length viral env glycoprotein genes were amplified and determined. Chimeric feline * human CD134 receptors were used to investigate the receptor utilization of 17 clones from Brazilian isolates of FIV. Analyses of the sequence present of molecular clones showed that all clones grouped within subtype B. In contrast to the virulent primary isolate FIV-GL8, expression of the first cysteine-rich domain (CRD1) of feline CD134 in the context of human CD134 was sufficient for optimal receptor function for all Brazilian FIV isolates tested. PMID- 21619903 TI - Functional improvement of an IRQ-PEG-MEND for delivering genes to the lung. AB - The targeted delivery of genes to endothelial cells is a potential strategy for curing certain types of disorders including cancer, inflammation and obesity. We previously reported that a liposome (IRQ-LP) modified with the IRQ peptide (IRQRRRR) was taken up by cells via a unique pathway, namely caveolar endocytosis, a cellular uptake pathway that is involved in the blood-to-tissue uptake of macromolecules in vascular endothelial cells. In the present study, we initally investigated the effect of IRQ peptide-modification on the biodistribution of poly(ethyleneglycol) (PEG)-coated liposomes (PEG-LP) after i.v. administration. The IRQ peptide-modified PEG-LP (IRQ-PEG-LP), as well as the PEG-LP were found to be mainly accumulated in the liver. Nevertheless, the fold increase in the lung accumulation of IRQ-PEG-LP, compared to the PEG-LP (approximately 20-folds) was substantially higher than other tissues (<5-fold). Thus, IRQ could function as a target ligand for lungs. We then used the IRQ peptide as a model for a ligand for targeting normal tissue endothelial cells, and then applied it to a gene delivery system. We previously developed a multifunctional envelope-type nano device (MEND), in which plasmid DNA is condensed using a polycation to form a core particle that is encapsulated in a lipid envelope. We modified the IRQ-modified PEG to the MEND (IRQ-PEG-MEND) and marker gene expression was evaluated after i.v. administration. However the transgene expression of the IRQ-PEG-MEND in lungs was low. This is most likely due to the inhibitory effect of the PEG spacer on intracellular trafficking (especially endosomal escape) of the IRQ-PEG-MEND. To overcome the dilemma associated with PEGylation, we improved the MEND system from the point of view of PEG length, lipid chain of the PEG derivative, the polycation and cationic lipid. As a result, transgene expression in lungs was enhanced in stepwise manner, and was finally improved by 5 orders of magnitude compared with the original IRQ-PEG MEND. Overcoming the dilemma of PEGylation is critical issue for in vivo applications of gene delivery targeting endothelial cells. PMID- 21619904 TI - Central mechanisms involved in the orexigenic actions of ghrelin. AB - Ghrelin is a stomach hormone, secreted into the bloodstream, that initiates food intake by activating NPY/AgRP neurons in the hypothalamic acruate nucleus. This review focuses on recent evidence that details the mechanisms through which ghrelin activate receptors on NPY neurons and downstream signaling within NPY neurons. The downstream signaling involves a novel CaMKK-AMPK-CPT1-UCP2 pathway that enhances mitochondrial efficiency and buffers reactive oxygen species in order to maintain an appropriate firing response in NPY. Recent evidence that shows metabolic status affects ghrelin signaling in NPY is also described. In particular, ghrelin does not activate NPY neurons in diet-induced obese mice and ghrelin does not increase food intake. The potential mechanisms and implications of ghrelin resistance are discussed. PMID- 21619905 TI - International seroepidemiology of adenovirus serotypes 5, 26, 35, and 48 in pediatric and adult populations. AB - Recombinant adenovirus serotype 5 (rAd5) vaccine vectors for HIV-1 and other pathogens have been shown to be limited by high titers of Ad5 neutralizing antibodies (NAbs) in the developing world. Alternative serotype rAd vectors have therefore been constructed. Here we report Ad5, Ad26, Ad35, and Ad48 NAb titers in 4381 individuals from North America, South America, sub-Saharan Africa, and Southeast Asia. As expected, Ad5 NAb titers were both frequent and high magnitude in sub-Saharan Africa and Southeast Asia. In contrast, Ad35 NAb titers proved infrequent and low in all regions studied, and Ad48 NAbs were rare in all regions except East Africa. Ad26 NAbs were moderately common in adults in sub-Saharan Africa and Southeast Asia, but Ad26 NAb titers proved markedly lower than Ad5 NAb titers in all regions, and these relatively low Ad26 NAb titers did not detectably suppress the immunogenicity of 4*10(10)vp of a rAd26-Gag/Pol/Env/Nef vaccine in rhesus monkeys. These data inform the clinical development of alternative serotype rAd vaccine vectors in the developing world. PMID- 21619906 TI - HPV vaccine information-seeking behaviors among US physicians: government, media, or colleagues? AB - The multiple information sources available may pose a challenge to physicians in providing accurate human papillomavirus (HPV) vaccine information to patients. The purpose of this study was to describe physicians' HPV vaccine information seeking behaviors and assess if these behaviors differ by physician specialty and sociodemographic characteristics. In 2009, 1008 Family Physicians (FPs), Pediatricians (Peds), and Obstetricians/Gynecologists (OBGYNs) completed a survey to assess their HPV vaccine information-seeking behaviors and vaccination practices. The largest proportion obtained HPV vaccine information from professional organizations (50.0%), followed by the Advisory Council on Immunization Practices (ACIP) (36.0%), and medical conferences (33.1%). Peds and FPs were more likely to obtain vaccine information from the ACIP (p-values<0.05). OBGYNs, non-White/Caucasian physicians, and those aged 40-49 were more likely to obtain vaccine information from internet websites (p-values<0.05). There is a need for targeted HPV vaccine communication approaches based on sociodemographic and physician specialty characteristics. PMID- 21619907 TI - Bis-(3',5')-cyclic dimeric adenosine monophosphate: strong Th1/Th2/Th17 promoting mucosal adjuvant. AB - New effective adjuvants are required to improve the performance of subunit vaccines. Here, we showed that bis-(3',5')-cyclic dimeric adenosine monophosphate (c-di-AMP), a second messenger molecule in bacteria and archaea, exerts strong adjuvant activities when delivered by mucosal route. In vitro studies showed that c-di-AMP was able to both stimulate pre-activated murine macrophages and promote the activation and maturation of dendritic cells of murine and human origin. Co administration of c-di-AMP with beta-galactosidase (beta-Gal) by intranasal route to BALB/c mice resulted in the elicitation of significantly higher serum antigen specific IgG titres than in controls. The induction of local immune responses was shown by the production of antigen-specific secretory IgA in different mucosal territories. In addition, strong cellular immune responses were observed against both the beta-Gal protein and a peptide encompassing its MHC class I-restricted epitope. The ratio of beta-Gal-specific antibodies and the secreted cytokine profiles by in vitro re-stimulated splenocytes suggested that a balanced Th1/Th2/Th17 response pattern is promoted by c-di-AMP. When C57BL/6 mice were immunized with OVA and c-di-AMP, vigorous in vivo CTL responses were also observed. These results indicated that c-di-AMP exhibits a high potential as adjuvant for the development of mucosal vaccines, in particular when cellular immunity is needed. PMID- 21619908 TI - Vaccination with epigenetically treated mesothelioma cells induces immunisation and blocks tumour growth. AB - Malignant mesothelioma (MM) is an aggressive tumour associated with poor outcome in patients. Current treatments for MM are of limited efficacy. Our recent findings suggest that epigenetic drugs may induce both cytotoxicity and an immune response against MM cells. Thus, we used a mouse model of MM (AK7) to analyse how epigenetic drugs could modulate MM development in vivo. The treatment of tumour bearing mice with an epigenetic drug already tested in clinical MM treatments (SAHA/Vorinostat) reduced the tumour mass and induced a moderate lymphocytic infiltration. However, the treatment did not stop tumour development. In order to show the potential effect of this epigenetic drug on tumour immunogenicity, in addition to cell cytotoxicity, we immunised mice either with AK7 cells pre treated with SAHA, or with one of two cytotoxic drugs (curcumin or selenite), prior to transplantation of live AK7 cells. A specific immune response was observed only in mice immunised with AK7 cells pre-treated with the epigenetic drug (SAHA) and the tumour growth was arrested. An increase in the proportion of CD3+ CD8+ lymphocytes occurred in the peritoneal cavity. We also observed large conglomerates of immune cells in the omentum with clusters of CD8+ T cells, together with lymphocytes directed against residual AK7 cells in the interlobular connective tissue of the pancreas. Our data demonstrate that epigenetic drugs, such as SAHA, can stimulate tumour immunogenicity and improve the recognition of aggressive MM cells by the immune system in vivo. PMID- 21619909 TI - A randomized, double-blind trial to evaluate immunogenicity and safety of 13 valent pneumococcal conjugate vaccine given concomitantly with trivalent influenza vaccine in adults aged >=65 years. AB - This randomized, double-blind study evaluated concomitant administration of 13 valent pneumococcal conjugate vaccine (PCV13) and trivalent inactivated influenza vaccine (TIV) in adults aged >=65 years who were naive to 23-valent pneumococcal polysaccharide vaccine. Patients (N=1160) were randomized 1:1 to receive PCV13+TIV followed by placebo, or Placebo+TIV followed by PCV13 at 0 and 1 months, with blood draws at 0, 1, and 2 months. Slightly lower pneumococcal serotype-specific anticapsular polysaccharide immunoglobulin G geometric mean concentrations were observed with PCV13+TIV relative to PCV13. Concomitant PCV13+TIV demonstrates acceptable immunogenicity and safety compared with either agent given alone. PMID- 21619910 TI - The uptake and intracellular fate of a series of different surface coated quantum dots in vitro. AB - Quantum dots (QDs) are potentially beneficial semi-conductor nanocrystals for use in diagnostics and therapeutics. The chemical composition of QDs however, has raised concerns as to their potential toxicity. Although a thorough examination using specific biochemical endpoints is necessary to assess QD toxicity, an understanding of the interaction of QDs, specifically their uptake and intracellular fate, with biological systems is also essential in determining their potential hazardous effects. The aim of this study was to investigate the uptake and intracellular fate of a series of different surface coated QDs (organic, carboxylated (COOH) and amino (NH2) polyethylene glycol (PEG)) in J774.A1 'murine macrophage-like' cells. Model 20 nm and 200 nm COOH polystyrene beads (PBs) were also studied. Results showed that COOH and NH2 (PEG) QDs, as well as 20 nm and 200 nm PBs were located within lysosomes and the mitochondria of macrophages after 2 h. Additionally, elemental transmission electron microscopy confirmed both COOH and NH2 (PEG) QDs to be located within membrane bound compartments at this time point. The data from this study combined with current knowledge, indicates that the intracellular localisation of QDs could be directly related to their toxicity. PMID- 21619911 TI - Special issue: integration of OMICs datasets into metabolic pathway analysis. PMID- 21619912 TI - Fast modulation of alpha activity during visual processing and motor control. AB - Whereas some studies suggest that alpha rhythm promotes information processing in the human cortex (processing hypothesis), other studies suggest its involvement in an active cortical idling which prevents the interfering action of irrelevant information (idling hypothesis). In this study, this apparent contradiction was analyzed using a computing procedure which distinguishes phase-locked and non phase-locked alpha response during the execution of a complex event-related visual-motor task. The electroencephalographies (EEGs) of 12 male volunteers were digitized (128 electrodes), band-pass filtered for isolating alpha wave, and event-related averaged during the execution of the visual-motor tasks. This procedure showed a phase-locked alpha response to stimuli and suppressed the non phase-locked response. When EEG envelope of the alpha-wave was computed (Hilbert transformation) before the event-related average, the response of the alpha amplitude to stimuli was observed while the phase-locked alpha response vanished. Visual stimuli induced a short-lasting increase of phase-locked alpha activity and a long-lasting decrease of non-phase-locked alpha activity whose latency and amplitude changed with the cortical region (visual vs. parietal vs. frontal cortex), with the sensory-semantic information of visual stimuli, and with the tasks associated to them (comparing the alpha response to stimuli which were used for a visual-motor tasks with those passively observed). Alpha sub-bands around the individual alpha frequency peak showed a different phase-locked response. Finally, two early evoked potentials (C1-P1) showed a time latency similar to that computed for the phase-locked alpha response, suggesting that early evoked potentials are modified by the superposition of this alpha activity. Present data suggest that alpha activity promotes the cortical processing of information by increasing the phase-locked alpha activity and by decreasing the non-phase-locked alpha activity, and disturbs cortical processing under basal conditions when it is not phase-locked to any particular task. PMID- 21619913 TI - Latrepirdine increases cerebral glucose utilization in aged mice as measured by [18F]-fluorodeoxyglucose positron emission tomography. AB - Latrepirdine is hypothesized to exert a unique mechanism of action involving stabilization of mitochondria that may have utility in treating Alzheimer's disease. However, the ability of latrepirdine to improve cognition in Alzheimer's disease (AD) is controversial due to a discrepancy between the positive signal reported in the multi-site phase II clinical trial where latrepirdine met all primary and secondary endpoints [Doody et al. (2008) Lancet 372:207-215], and the subsequent null effect observed in a multicenter, phase III trial. While dysfunction of mitochondria and abnormal energy metabolism has been linked to AD pathology, no studies have been reported that investigate latrepirdine's effect on cerebral glucose utilization (CGU). Glucose metabolism, following acute latrepirdine administration, can be used to help dose selection in Phase I dose ranging studies. The aim of the current study was to assess changes in CGU in young and aged mice in vivo using [18F]-fluorodeoxyglucose positron emission tomography (FDG-PET) after acute treatment with latrepirdine. Two ages of B6SJLF2 mice (5 and 20 months old) were tested. Three test-retest FDG-PET baseline scans were assessed across all subjects. As CGU was heterogeneous in aged mice, compared to young mice, aged subjects were rank ordered and then counterbalanced into two CGU homogenous groups. In Studies 1 and 2, latrepirdine (1.0 mg/kg) significantly enhanced CGU in aged mice. In contrast, Study 3 revealed that latrepirdine did not modulate CGU in young mice. Monitoring changes in CGU in response to acute drug administration may represent an imaging biomarker for dose selection in AD. Further studies that would establish the translation from mice to non-human primates to humans need to be investigated to confirm the utility of FDG-PET in dose-selection for mitochondrial modulators. PMID- 21619914 TI - Intracerebroventricularly administered lipopolysaccharide enhances spike-wave discharges in freely moving WAG/Rij rats. AB - Peripheral lipopolysaccharide (LPS) injection enhances spike-wave discharges (SWDs) in the genetic rat model of absence epilepsy (Wistar Albino Glaxo/Rijswijk rats: WAG/Rij rats) parallel with the peripheral proinflammatory cytokine responses. The effect of centrally administered LPS on the absence-like epileptic activity is not known, however despite the important differences in inflammatory mechanisms. To examine the effect of centrally administered LPS on the pathological synchronization we intracerebroventricularly (i.c.v.) injected LPS into WAG/Rij rats and measured the number and duration of SWDs. I.c.v. injected LPS increased the number and duration of SWDs for 3h, thereafter, a decrease in epileptic activity was observed. To further investigate the nature of this effect, a non-steroid anti-inflammatory drug (indomethacin; IND) or a competitive N-methyl-d-aspartate (NMDA) receptor antagonist (2-amino-5-phosphonopentanoic acid; AP5) was injected intraperitoneally (i.p.), preceding the i.c.v. LPS treatment. IND abolished the i.c.v. LPS induced changes in SWDs, while AP5 extended it for 5h. As control treatments, both IND and AP5 application by themselves decreased the number of SWDs for 2 and 3h, respectively. Our results show that centrally injected LPS, likewise the peripheral injection, can increase the number and duration of SWDs in the WAG/Rij rat, and the effect invoke inflammatory cytokines as well as excitatory neurotransmitters. PMID- 21619915 TI - Physicochemical and pharmacokinetic characterization of a spray-dried malotilate emulsion. AB - Malotilate (MT) is a hepatoprotective drug administered orally. However, MT was found to be a poorly water-soluble drug with low oral bioavailability. In the present investigation, a novel spray-dried emulsion (SDE) loaded with MT was prepared, and its physicochemical properties were characterized by rheological evaluation, particle size measurement, in vitro release, and surface morphology. The pharmacokinetic study of SDE, in comparison to MT suspension with the pure MT powder homogeneously dispersed in 0.5% CMC-Na solution, was also performed in rats after a single oral dose. It was found that SDE exhibited a 2.9-fold higher peak plasma concentration (C(max)) and 2.3-fold higher area under the curve (AUC) than MT suspension. PMID- 21619916 TI - The characterization of protein release from sericin film in the presence of an enzyme: towards fibroblast growth factor-2 delivery. AB - Aqueous preparations of silk protein (sericin) films were prepared to evaluate their biodegradation properties. In the absence of trypsin, sericin film swelled rapidly, kept its shape, and remained unaltered for 28 days or longer due to form beta-sheet structures. In the presence of trypsin, sericin film gradually degraded; since the rate depended on the concentration of trypsin, the films likely underwent enzymatic hydrolysis. Sericin film incorporating the model protein drug fluorescein isothiocyanate-albumin (FA) also gradually degraded in the presence of trypsin and resulted in the sustained release of FA for 2 weeks or longer; in contrast, FA release was quite slow in the absence of trypsin. It is expected that sericin film has potential as a biodegradable and drug-releasing carrier. To evaluate the practical applicability of sericin film for the repair of defective tissues, fibroblast growth factor-2 (FGF-2) was incorporated into sericin films and the films were implanted on skull defects in rats. Whereas FGF 2 release was suppressed in the absence of trypsin in vitro, it appears that FGF 2, immobilized by ionic interactions between sericin and FGF-2, can be sustained released in vivo from films incorporating 2500 or 250 ng of FGF-2 to support the growth of tissue around wounds. PMID- 21619917 TI - Double-blinded randomized controlled trial for immunomodulatory effects of Tulsi (Ocimum sanctum Linn.) leaf extract on healthy volunteers. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Tulsi (Ocimum sanctum Linn.) is considered as a sacred herb and traditionally it is believed that consumption of Tulsi leaf on empty stomach increases immunity. Experimental studies have shown that alcoholic extract of Tulsi modulates immunity. MATERIALS AND METHODS: The present study was designed to evaluate the immunomodulatory effects of ethanolic extract of Tulsi leaves through a double-blinded randomized controlled cross-over trial on healthy volunteers. Three hundred milligrams capsules of ethanolic extracts of leaves of Tulsi or placebo were administered to 24 healthy volunteers on empty stomach and the results of 22 subjects who completed the study were analyzed. The primary objective was to study the levels of Th1 and Th2 cytokines (interferon-gamma and interleukin-4) during both pre and post intervention period in blood culture supernatants following stimulation with lipopolysaccharide and phytohaemagglutinin. Other immunological parameters such as T-helper and T cytotoxic cells, B-cells and NK-cells also were analyzed using Flowcytometry. RESULTS: Statistically significant increase in the levels of IFN-gamma (p=0.039), IL-4 (p=0.001) and percentages of T-helper cells (p=0.001) and NK-cells (p=0.017) were observed after 4 weeks in the Tulsi extract intervention group in contrast to the placebo group. CONCLUSIONS: These observations clearly ascertain the immunomodulatory role of Tulsi leaves extract on healthy volunteers. PMID- 21619918 TI - An aqueous birch leaf extract of Betula pendula inhibits the growth and cell division of inflammatory lymphocytes. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Leaf extracts of Betula pendula have been traditionally used for the treatment of patients with rheumatoid arthritis (RA) or osteoarthritis. AIM OF THE STUDY: We investigated the anti-proliferative capacity of an aqueous leaf extract of Betula pendula (BPE) on human primary lymphocytes in vitro, because activated lymphocytes play a major role in the initiation and maintenance of RA. MATERIALS AND METHODS: Lymphocyte proliferation and cell division was measured by the activity of mitochondrial dehydrogenases and by using the membrane-permeable dye carboxyfluorescein diacetate succinimidyl ester (CFSE), respectively. Apoptosis was analyzed by surface staining of phosphatidylserine and intracellular activation of effector caspases 3 and 7 in comparison to the drug methotrexate using flow cytometric and photometrical analysis. In addition, the impact of the extract on cell cycle distribution was investigated by propidium iodide staining of DNA. For the bioassays BPE concentrations of 10-160 MUg/mL were investigated. A phytochemical analysis, using LC-MS and HPLC, was conducted to identify the polyphenolic constituents of the birch leaf extract. RESULTS: Leaf extracts of Betula pendula inhibited the growth and cell division (CD8(+): 40 MUg/mL: 45%; 80 MUg/mL: 60%; 160 MUg/mL: 87%) (CD4(+): 40 MUg/mL: 33%; 80 MUg/mL: 54%; 160 MUg/mL: 79%) of activated, but not of resting T lymphocytes in a significant dose-dependent manner. The inhibition of lymphocyte proliferation due to apoptosis induction (compared to untreated control: 40 MUg/mL: 163%; 80 MUg/mL: 240%; 160 MUg/mL: 348%) and cell cycle arrest was comparable to that of methotrexate. LC-MS analyses showed that the extract contains different quercetin-glycosides. CONCLUSION: Our results give a rational basis for the use of Betula pendula leaf extract for the treatment of immune disorders, like rheumatoid arthritis, by diminishing proliferating inflammatory lymphocytes. PMID- 21619919 TI - Effects of Panax notoginseng saponins on proliferation and apoptosis of vascular smooth muscle cells. AB - AIM OF THE STUDY: Atherosclerosis is a common cardiovascular disease, and linked with the development of many cardiovascular complications, such as myocardial ischemia and stroke. Although pathogenesis of atherosclerosis is not completely elucidated, increasing evidence has demonstrated that abnormal proliferation of vascular smooth muscle cells (VSMCs) plays an important role in formation of atherosclerosis. Previous studies showed that saponins from Panax notoginseng (PNS) possess anti-atherosclerotic properties. However, the mechanism of PNS against atherosclerosis is not well understood. Therefore, the present study observed the effects of PNS on proliferation and apoptosis of VSMCs. MATERIALS AND METHODS: Rat VSMCs were cultured, and platelet-derived growth factor (PDGF) was used to stimulate cell proliferation. The viability of VSMCs was assessed with the MTT method. VSMCs apoptosis was detected by flow cytometry. Expressions of apoptosis related protein p53, Bax, caspase-3 and Bcl-2 were determined using Western blot. RESULTS: Pretreatment of the cells with PNS (200, 400, 800 MUg/mL) significantly inhibited proliferation of PDGF-stimulated VSMCs, and induced apoptosis of the proliferated VSMCs in a concentration-dependent way. Western blot analysis showed that PNS upregulated expressions of pro-apoptotic protein p53, Bax and caspase-3, downregulated expression of anti-apoptotic protein Bcl-2, and enlarged Bax/Bcl-2 ratio in the proliferated VSMCs induced by PDGF. CONCLUSIONS: This study demonstrates that PNS both inhibits VSMCs proliferation and induces VSMCs apoptosis through upregulating p53, Bax, caspase-3 expressions and downregulating Bcl-2 expression, which constitute the pharmacological basis of its anti-atherosclerotic action. PMID- 21619920 TI - Panax notoginseng saponins inhibit ischemia-induced apoptosis by activating PI3K/Akt pathway in cardiomyocytes. AB - AIM OF THIS STUDY: The panax notoginseng saponins (PNS) have been clinically used for the treatment of cardiovascular diseases and stroke in China. Evidences demonstrated that PNS could protect cardiomyocytes from injury induced by ischemia, but the underlying molecular mechanisms of this protective effect are still unclear. This study was aimed to investigate the protective effect and potential molecular mechanisms of PNS on apoptosis in H9c2 cells in vitro and rat myocardial ischemia injury model in vivo. MATERIALS AND METHODS: H9c2 cells subjected to serum, glucose and oxygen deprivation (SGOD) were used as in vitro models and SD rats subjected to left anterior descending (LAD) coronary artery ligation were used as in vivo models. The anti-apoptotic effect of PNS was evaluated by Annexin V/PI analysis or TUNEL assay. Mitochondrial membrane potential (Deltapsim) was detected by JC-1 analysis. The expression of Akt and phosphorylated Akt (p-Akt) were detected by western blot assay. RESULTS: PNS exhibited anti-apoptotic effect both in H9c2 cells and in ischemic myocardial tissues. However, the effect was blocked in vitro by LY294002, a specific PI3K inhibitor. The anti-apoptotic effect of PNS was mediated by stabilizing Deltapsim in H9c2 cells. Furthermore the indices of the left ventricular ejection fractions (EF), left ventricular fractional shortening (FS), left ventricular dimensions at end diastole (LVDd) and left ventricular dimensions at end systole (LVDs) suggested that PNS improved rats cardiac function. PNS significantly increased p Akt both in H9c2 cells and in ischemic myocardial tissues and this effect was also blocked by LY294002 in H9c2 cells. CONCLUSION: Results of this study suggested that PNS could protect myocardial cells from apoptosis induced by ischemia in both the in vitro and in vivo models through activating PI3K/Akt signaling pathway. PMID- 21619921 TI - Korean red ginseng (Panax ginseng C.A. Meyer) root fractions: differential effects on postprandial glycemia in healthy individuals. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Variations in ginsenoside profile may predict the postprandial glucose (PPG)-lowering efficacy of ginseng. Previously we reported differential PPG-lowering effects with two Korean red ginseng (KRG) root. FRACTIONS: body and rootlets, of variable ginsenoside profiles. Whether this effect is reproducible with a different KRG source is unclear. We therefore tested two root fractions from a KRG source with elevated ginsenoside levels to assess its effect on PPG. MATERIALS AND METHODS: After a 12-h overnight fast, 13 healthy individuals (6M:7F; age=28 +/- 10 y; BMI=24.1 +/- 3 kg/m2; FBG=4.77 +/- 0.04 mmol/L) randomly received either 3g of KRG-body, rootlets or placebo, on three separate visits. Treatments were consumed 60 min prior to a standard test meal with capillary blood samples at -60, 0, 15, 30, 45, 60, 90 and 120 min. RESULTS: The KRGrootlets had>6 fold total ginsensosides than the KRG-body but did not significantly affect PPG. Despite a reduced ginsenoside profile, KRG-body lowered PPG levels at 45, 60, 90 and 120 min during the test (p<0.05), rendering an overall reduction of 27% in incremental area under the glucose curve compared to the control (p<0.05). CONCLUSIONS: Comparing the results with a previously studied batch of KRG suggests a potential therapeutic dose range for ginsenosides. This observation should be clinically verified with acute screening and ginsenoside composition analysis. PMID- 21619922 TI - The anti-inflammatory activities of an extract and compounds isolated from Platycladus orientalis (Linnaeus) Franco in vitro and ex vivo. AB - ETHNOPHARMACOLOGICAL RELEVANCE: As a Chinese traditional herbal medicine, leaves of Platycladus orientalis (Linnaeus) Franco (LPO) are used to treat coughs, excessive mucus secretion, chronic bronchitis, bronchiectasis, and asthma, etc. The experiments were carried out to investigate their anti-inflammatory properties and mechanisms, which could support the Chinese traditional uses of treating inflammatory airway diseases. MATERIALS AND METHODS: The anti inflammatory activities of the chloroform fraction (CHL) and pure compounds of LPO were evaluated for their abilities to inhibit pro-inflammatory enzymes in vitro, and production of tumor necrosis factor-alpha (TNF-alpha) and nitric oxide in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Furthermore, the arachidonic acid metabolites, stimulated by calcium ionophore A23187, were also determined by HPLC. RESULTS: For the first time, the assays of eicosanoids in intact cells showed that the CHL, hinokiol, and acacetin had significant inhibitory effects on 5-hydroxy-eicosa-tetra-enoic acid (5-HETE) and leukotriene B(4) (LTB4) formations. And cell-free enzyme assays (5-lipoxygenase, leukotriene A(4)-hydrolase, cyclooxgenase-2) demonstrated the potent inhibitory effects of the CHL, hinokiol and acacetin on 5-lipoxygenase (5-LOX). Then, the inhibitions of the CHL, hinokiol on NO biosynthesis and the inhibitions of the CHL, 8(14),15 pimaradien-3beta,18-diol, and hinokiol on TNF-alpha release were also confirmed in the RAW264.7 murine macrophages. CONCLUSION: The data indicate that the inhibitory effects of the CHL and its components (hinokiol and acacetin) on 5-LOX contribute to the anti-inflammatory activity of LPO. Moreover, the CHL and its components also show beneficial effects on NO and TNF-alpha production. Consequently, these results provide a rationale for LPO's traditional applications in the treatment of inflammatory airway diseases. PMID- 21619923 TI - The memory-enhancing effects of Kami-ondam-tang in mice. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Kami-ondam-tang (KOT), a traditional Chinese medicine, has been used to treat mental and neuropsychiatric disorders, including dementia. This study aimed to investigate the effects of KOT on cognition and the mechanisms underlying these effects in mice. MATERIALS AND METHODS: Using the passive avoidance task, we investigated the effect of sub-chronic administration of KOT on the cognition of mice. We also examined the expressions of protein kinase B (Akt), cAMP response element-binding protein (CREB), brain-derived neurotrophic factor (BDNF), and doublecortin (DCX) in the hippocampal CA1 and dentate gyrus regions using immunohistochemistry and western blotting. RESULTS: The administration of KOT (50mg/kg/day, p.o.) for 14 days significantly increased step-through latency in the passive avoidance task compared with vehicle-treated controls. Furthermore, KOT administration (50mg/kg/day, p.o.) significantly increased the expressions of phosphorylated Akt, phosphorylated CREB and BDNF in the hippocampal CA1 and dentate gyrus. In addition, KOT administration resulted in a significant increase in the number of DCX-immunopositive cells in the dentate gyrus. CONCLUSIONS: These results suggest that KOT enhances cognitive performance through the upregulation of Akt-CREB-BDNF signaling and neurogenesis. PMID- 21619924 TI - A comparison of the immunostimulatory effects of the medicinal herbs Echinacea, Ashwagandha and Brahmi. AB - AIM OF THE STUDY: Herbs, as food or medicine, can strengthen the body and increase its resistance to illnesses by acting on various components of the immune system. For example, Echinacea is noted for its ability to enhance immune function, primarily through activation of the innate immune responses. Here, we investigated the potential for two herbs commonly found in India, Ashwagandha (Withania somnifera) and Brahmi (Bacopa monnieri), to enhance immune function and compared their effects to that of Echinacea. MATERIALS AND METHODS: Sprague Dawley rats were fed a diet supplemented with 1% (w/w) Echinacea, Ashwagandha, or Brahmi for 4 weeks to examine their effects on immune function. RESULTS: The Brahmi diet stimulated more secretion of IgA and IgG in the serum compared to Echinacea or Ashwagandha. Whether or not lectin was present in the diet, the production of IgA, IgG and IgM in spleen lymphocytes increased with herbal supplements. The concentrations of IFN-gamma and IL-2 treated with LPS and ConA were significantly higher in the dietary herb than in the control. On the contrary, TNF-alpha production in rats receiving dietary herbal supplements was significantly lower compared to the control animals. CONCLUSION: Herbal remedies based on Echinacea, Brahmi, or Ashwagandha can enhance immune function by increasing immunoglobulin production. Furthermore, these herbal medicines might regulate antibody production by augmenting both Th1 and Th2 cytokine production. PMID- 21619925 TI - Oxidative stress in the kidney of reproductive male rats during aging. AB - Reproduction alters the male physiology. We performed a comprehensive examination of oxidative stress in the kidneys of male rats with (experienced) or without (naive) reproductive activity during aging. Oxidative stress was assessed by measuring the activity of catalase, glutathione peroxidase, glutathione S transferase, and superoxide dismutase, and by measuring protein carbonylation, lipid peroxidation, nitrite and nitrate levels, vitamin C levels, and glutathione (total, reduced, and oxidized forms) levels, and metabolism was accessed by aconitase activity in kidney tissue, as well as testosterone and estradiol levels in serum. Reproductively active animals exhibited increased testosterone levels and altered metabolism. Aging affects tissues and organs and contributes to their functional decline. Elderly naive rats showed high nitrite and nitrate levels. The experienced rats had less damage in elderly ages, probably because they had higher antioxidant amount and antioxidant enzyme activities at earlier ages, which would have avoided oxidative damage seen in naive group, and because of the metabolism decline. Glutathione increase in naive elder rats probably was induced for direct protection against oxidative damage and indirect protection by higher glutathione peroxidase and glutathione S-transferase activities. Linear regression shows that lipid peroxidation levels explained vitamin C levels (B standardized value of 0.42), indicating that vitamin C was properly produced or recruited into kidneys to combat lipid peroxidation. Catalase activity reflected the protein carbonylation and lipid peroxidation levels (B standardized values of 0.28 and 0.48). These results add comprehensive data regarding changes in oxidative stress during aging, and suggest an explanation for the costs of reproduction. PMID- 21619926 TI - Advances and challenges in analytical characterization of biotechnology products: mass spectrometry-based approaches to study properties and behavior of protein therapeutics. AB - Biopharmaceuticals are a unique class of medicines due to their extreme structural complexity. The structure of these therapeutic proteins is critically important for their efficacy and safety, and the ability to characterize it at various levels (from sequence to conformation) is critical not only at the quality control stage, but also throughout the discovery and design stages. Biological mass spectrometry (MS) offers a variety of approaches to study structure and behavior of complex protein drugs and has already become a default tool for characterizing the covalent structure of protein therapeutics, including sequence and post-translational modifications. Recently, MS-based methods have also begun enjoying a dramatic growth in popularity as a means to provide information on higher order structure and dynamics of biotechnology products. In particular, hydrogen/deuterium exchange MS and charge state distribution analysis of protein ions in electrospray ionization (ESI) MS offer a convenient way to assess the integrity of protein conformation. Native ESI MS also allows the interactions of protein drugs with their therapeutic targets and other physiological partners to be monitored using simple model systems. MS-based methods are also applied to study pharmacokinetics of biopharmaceutical products, where they begin to rival traditional immunoassays. MS already provides valuable support to all stages of development of biopharmaceuticals, from discovery to post-approval monitoring, and its impact on the field of biopharmaceutical analysis will undoubtedly continue to grow. PMID- 21619927 TI - Oral delivery of curcumin bound to chitosan nanoparticles cured Plasmodium yoelii infected mice. AB - Curcumin has been shown to have anti malarial activity, but poor bioavailability and chemical instability has hindered its development as a drug. We have bound curcumin to chitosan nanoparticles to improve its bioavailability and chemical stability. We found that curcumin bound to chitosan nanoparticles did not degrade that rapidly in comparison to free curcumin when such particles were incubated in mouse plasma in vitro at room temperature. The uptake of bound curcumin from chitosan nanoparticles by mouse RBC was much better than from free curcumin. Oral delivery of curcumin bound chitosan nanoparticles to normal mice showed that they can cross the mucosal barrier intact and confocal microscopy detected the nanoparticles in the blood. Curcumin loaded chitosan nanoparticles when delivered orally improved the bioavailability of curcumin in the plasma and RBC. While mice infected with a lethal strain of Plasmodium yoelii (N-67) died between 8 and 9 days post infection, feeding of chitosan nanoparticles alone made them to survive for five more days. Feeding 1mg of native curcumin to infected mice per day for seven days resulted in survival of one third of mice but under the same condition when 1mg of curcumin bound to chitosan nanoparticles was fed all the mice survived. Like chloroquine, curcumin inhibited parasite lysate induced heme polymerization in vitro in a dose dependent manner and curcumin had a lower IC(50) value than chloroquine. We believe that binding of curcumin to chitosan nanoparticles increases its chemical stability and enhances its bioavailability when fed to mice. In vitro data suggest that it can inhibit hemozoin synthesis which is lethal for the parasite. PMID- 21619928 TI - Extending life span by increasing oxidative stress. AB - Various nutritional, behavioral, and pharmacological interventions have been previously shown to extend life span in diverse model organisms, including Saccharomyces cerevisiae, Caenorhabditis elegans, Drosophila melanogaster, mice, and rats, as well as possibly monkeys and humans. This review aims to summarize published evidence that several longevity-promoting interventions may converge by causing an activation of mitochondrial oxygen consumption to promote increased formation of reactive oxygen species (ROS). These serve as molecular signals to exert downstream effects to ultimately induce endogenous defense mechanisms culminating in increased stress resistance and longevity, an adaptive response more specifically named mitochondrial hormesis or mitohormesis. Consistently, we here summarize findings that antioxidant supplements that prevent these ROS signals interfere with the health-promoting and life-span-extending capabilities of calorie restriction and physical exercise. Taken together and consistent with ample published evidence, the findings summarized here question Harman's Free Radical Theory of Aging and rather suggest that ROS act as essential signaling molecules to promote metabolic health and longevity. PMID- 21619931 TI - Mutations in an amino acid transporter gene are responsible for sex-linked translucent larval skin of the silkworm, Bombyx mori. AB - The sex-linked translucent (os) mutation in the silkworm, Bombyx mori, confers slightly translucent larval skin resulting from a decrease in the incorporation of uric acid into epidermal cells. By positional cloning, we narrowed a region linked to the os phenotype to approximately 157 kb located on scaffold Bm_scaf72 on the Z chromosome (chromosome 1). The region contained four gene models. Sequencing analysis revealed that one of the candidate genes had a 7-bp deletion in the coding region. We also found a 111-bp deletion or single-nucleotide substitution in the same gene using independent os mutant strains. Because all the mutations caused the generation of abnormal transcripts followed by translation of a truncated protein, we conclude that the mutation of this candidate gene is responsible for the translucent larval skin of the os mutant. Sequence analysis indicated that the gene responsible for the os mutation had homology to amino acid transporters of the solute carrier family of proteins. Our results suggest that solute carrier proteins are involved in uric acid transport in insects and other invertebrates. PMID- 21619930 TI - Implication of the oligomeric state of the N-terminal PTX3 domain in cumulus matrix assembly. AB - Pentraxin 3 (PTX3) plays a key role in the formation of the hyaluronan-rich matrix of the cumulus oophorus surrounding ovulated eggs that is required for successful fertilization and female fertility. PTX3 is a multimeric protein consisting of eight identical protomers held together by a combination of non covalent interactions and disulfide bonds. Recent findings suggest that the oligomeric status of PTX3 is important for stabilizing the cumulus matrix. Because the role of PTX3 in the cumulus resides in the unique N-terminal sequence of the protomer, we investigated further this issue by testing the ability of distinct Cys/Ser mutants of recombinant N-terminal region of PTX3 (N(_)PTX3) with different oligomeric arrangement to promote in vitro normal expansion in cumuli from Ptx3-null mice. Here we report that the dimer of the N(_)PTX3 is unable to rescue cumulus matrix organization, and that the tetrameric assembly of the protein is the minimal oligomeric state required for accomplishing this function. We have previously demonstrated that PTX3 binds to HCs of IalphaI and TSG-6, which are essential for cumulus matrix formation and able to interact with hyaluronan. Interestingly, here we show by solid-phase binding experiments that the dimer of the N(_)PTX3 retains the ability to bind to both IalphaI and TSG-6, suggesting that the octameric structure of PTX3 provides multiple binding sites for each of these ligands. These findings support the hypothesis that PTX3 contributes to cumulus matrix organization by cross-linking HA polymers through interactions with multiple HCs of IalphaI and/or TSG-6. The N-terminal PTX3 tetrameric oligomerization was recently reported to be also required for recognition and inhibition of FGF2. Given that this growth factor has been detected in the mammalian preovulatory follicle, we wondered whether FGF2 negatively influences cumulus expansion and PTX3 may also serve in vivo to antagonize its activity. We found that a molar excess of FGF2, above PTX3 binding capacity, does not affect in vitro cumulus matrix formation thus ruling out this possibility. In conclusion, the data strength the view that PTX3 acts as a nodal molecule in cross-linking HA in the matrix. PMID- 21619932 TI - Practical performance evaluation of a 10k * 10k CCD for electron cryo-microscopy. AB - Electron cryo-microscopy (cryo-EM) images are commonly collected using either charge-coupled devices (CCD) or photographic film. Both film and the current generation of 16 megapixel (4k * 4k) CCD cameras have yielded high-resolution structures. Yet, despite the many advantages of CCD cameras, more than two times as many structures of biological macromolecules have been published in recent years using photographic film. The continued preference to film, especially for subnanometer-resolution structures, may be partially influenced by the finer sampling and larger effective specimen imaging area offered by film. Large format digital cameras may finally allow them to overtake film as the preferred detector for cryo-EM. We have evaluated a 111-megapixel (10k * 10k) CCD camera with a 9 MUm pixel size. The spectral signal-to-noise ratios of low dose images of carbon film indicate that this detector is capable of providing signal up to at least 2/5 Nyquist frequency potentially retrievable for 3D reconstructions of biological specimens, resulting in more than double the effective specimen imaging area of existing 4k * 4k CCD cameras. We verified our estimates using frozen-hydrated epsilon15 bacteriophage as a biological test specimen with previously determined structure, yielding a ~7 A resolution single particle reconstruction from only 80 CCD frames. Finally, we explored the limits of current CCD technology by comparing the performance of this detector to various CCD cameras used for recording data yielding subnanometer resolution cryo-EM structures submitted to the electron microscopy data bank (http://www.emdatabank.org/). PMID- 21619929 TI - Inorganic nitrite therapy: historical perspective and future directions. AB - Over the past several years, investigators studying nitric oxide (NO) biology and metabolism have come to learn that the one-electron oxidation product of NO, nitrite anion, serves as a unique player in modulating tissue NO bioavailability. Numerous studies have examined how this oxidized metabolite of NO can act as a salvage pathway for maintaining NO equivalents through multiple reduction mechanisms in permissive tissue environments. Moreover, it is now clear that nitrite anion production and distribution throughout the body can act in an endocrine manner to augment NO bioavailability, which is important for physiological and pathological processes. These discoveries have led to renewed hope and efforts for an effective NO-based therapeutic agent through the unique action of sodium nitrite as an NO prodrug. More recent studies also indicate that sodium nitrate may also increase plasma nitrite levels via the enterosalivary circulatory system resulting in nitrate reduction to nitrite by microorganisms found within the oral cavity. In this review, we discuss the importance of nitrite anion in several disease models along with an appraisal of sodium nitrite therapy in the clinic, potential caveats of such clinical uses, and future possibilities for nitrite-based therapies. PMID- 21619933 TI - From SPINE to SPINE-2 complexes and beyond. PMID- 21619934 TI - Development of a diagnostic rule for identifying radiographic osteoarthritis in people with first metatarsophalangeal joint pain. AB - OBJECTIVE: To develop a diagnostic rule for the identification of radiographic osteoarthritis (OA) of the first metatarsophalangeal joint (MTPJ) in people with first MTPJ pain. DESIGN: Symptoms and clinical observations were documented in 181 people with first MTPJ pain, and the presence of OA was confirmed using plain film radiography. Diagnostic test statistics were calculated to assess the ability of symptoms and clinical observations to identify radiographic OA. Multivariate logistic regression was used to develop two diagnostic models: a statistically optimal model and a simplified clinical model. RESULTS: Multivariate logistic regression identified pain duration greater than 25 months, the presence of a dorsal exostosis, hard-end feel, crepitus and less than 64 degrees of first MTPJ dorsiflexion to be significantly associated with radiographic OA. The statistically optimal model and clinical model performed similarly, with the areas under the receiver operating characteristics curves being 0.87 (95% confidence interval [CI] 0.81-0.93) and 0.87 (95% CI 0.80-0.93), respectively, and the percentage of cases correctly classified being 86.2 and 85.6, respectively. A cut-off score of >=3 using the clinical model resulted in a sensitivity of 88%, specificity of 71%, accuracy of 84%, positive likelihood ratio of 3.07 and negative likelihood ratio of 0.17. CONCLUSIONS: In people with first MTPJ pain, a model consisting of five clinical observations can accurately identify the presence or absence of radiographic OA. The application of this diagnostic rule may assist clinical decision making and potentially reduce the need for referral for radiographs. PMID- 21619935 TI - Increased chondrocyte sclerostin may protect against cartilage degradation in osteoarthritis. AB - OBJECTIVES: To investigate the regulation of sclerostin (SOST) in osteoarthritis (OA) and its potential effects on articular cartilage degradation. METHODS: SOST and other Wnt-beta-catenin components were immuno-localised in osteochondral sections of surgically-induced OA in knees of sheep and mice, and human OA samples obtained at arthroplasty. Regulation of SOST mRNA and protein expression by ovine chondrocytes in response to interleukin-1alpha (IL-1alpha) or tumour necrosis factor-alpha (TNFalpha) was examined in explant cultures. The effect of 25 or 250 ng/ml recombinant SOST alone or in combination with IL-1alpha, on ovine articular cartilage explant aggrecan degradation, and chondrocyte gene expression of Wnt-beta-catenin pathway proteins, metalloproteinases and their inhibitors, and cartilage matrix proteins was quantified. RESULTS: Contrary to being an osteocyte-specific protein, SOST was expressed by articular chondrocytes, and mRNA levels were upregulated in vitro by IL-1alpha but not TNFalpha. Chondrocyte SOST staining was significantly increased only in the focal area of cartilage damage in surgically-induced OA in sheep and mice, as well as end-stage human OA. In contrast, osteocyte SOST was focally decreased in the subchondral bone in sheep OA in association with bone sclerosis. SOST was biologically active in chondrocytes, inhibiting Wnt-beta-catenin signalling and catabolic metalloproteinase [matrix metalloproteinases (MMP) and distintegrin and metalloproteinase with thrombospndin repeats (ADAMTS)] expression, but also decreasing mRNA levels of aggrecan, collagen II and tissue inhibitors of metalloproteinaes (TIMPs). Despite this mixed effect, SOST dose-dependently inhibited IL-1alpha-stimulated cartilage aggrecanolysis in vitro. CONCLUSIONS: These results implicate SOST in regulating the OA disease processes, but suggest opposing effects by promoting disease-associated subchondral bone sclerosis while inhibiting degradation of cartilage. PMID- 21619936 TI - Acute joint pathology and synovial inflammation is associated with increased intra-articular fracture severity in the mouse knee. AB - OBJECTIVE: Post-traumatic arthritis is a frequent cause of disability and occurs most commonly and predictably after articular fracture. The objective of this investigation was to examine the effect of fracture severity on acute joint pathology in a novel murine model of intra-articular fracture. DESIGN: Low and high energy articular fractures (n=25 per group) of the tibial plateau were created in adult male C57BL/6 mice. The acute effect of articular fracture severity on synovial inflammation, bone morphology, liberated fracture area, cartilage pathology, chondrocyte viability, and systemic cytokines and biomarkers levels was assessed at 0, 1, 3, 5, and 7 days post-fracture. RESULTS: Increasing intra-articular fracture severity was associated with greater acute pathology in the synovium and bone compared to control limbs, including increased global synovitis and reduced periarticular bone density and thickness. Applied fracture energy was significantly correlated with degree of liberated cortical bone surface area, indicating greater comminution. Serum concentrations of hyaluronic acid (HA) were significantly increased 1 day post-fracture. While articular fracture significantly reduced chondrocyte viability, there was no relationship between fracture severity and chondrocyte viability, cartilage degeneration, or systemic levels of cytokines and biomarkers. CONCLUSIONS: This study demonstrates that articular fracture is associated with a loss of chondrocyte viability and increased levels of systemic biomarkers, and that increased intra-articular fracture severity is associated with increased acute joint pathology in a variety of joint tissues, including synovial inflammation, cortical comminution, and bone morphology. Further characterization of the early events following articular fracture could aid in the treatment of post-traumatic arthritis. PMID- 21619937 TI - NSAIDs vs acetaminophen in knee and hip osteoarthritis: a systematic review regarding heterogeneity influencing the outcomes. AB - OBJECTIVE: To identify sources of heterogeneity (statistical, methodological, and clinical) in studies evaluating non-steroidal anti-inflammatory drugs (NSAIDs) vs acetaminophen in patients with knee and hip osteoarthritis (OA) to elucidate variations in outcomes. METHOD: A database search (1966 to January 2010) was made for (randomized) controlled trials ((R)CTs) comparing NSAIDs vs acetaminophen in knee and hip OA. Extracted data included baseline demographic/clinical characteristics, outcomes at follow-up, and characteristics of study design. Heterogeneity was examined with subgroup analyses by exploring changes in effect size and with I(2) of Higgins. Pain measures were expressed as standardized mean differences. RESULTS: 15 RCTs, including 21 comparisons of NSAIDs and acetaminophen were included. Statistical heterogeneity was absent (Cochran's Q test=14.11; I(2)=0; P=0.78). Moderate clinical heterogeneity was found for comparisons which included both hip and knee OA vs knee OA only (I(2)=51; P=0.09). NSAIDs seemed slightly more effective than acetaminophen if more patients with hip OA were included. However, the pooled effect sizes of comparisons with knee OA vs both knee and hip OA are equal. Low clinical heterogeneity was found for comparisons with low dosage of acetaminophen, normal dosage of NSAIDs, and moderate pain intensity at baseline. Low methodological heterogeneity was found for comparisons with a short duration. CONCLUSION: Future trials should present the results of hip and knee OA separately, as moderate clinical heterogeneity was found. There might be differences in effectiveness of NSAIDs vs acetaminophen in patients with hip vs knee OA. No significant methodological and statistical heterogeneity was found in studies evaluating NSAIDs vs acetaminophen. PMID- 21619938 TI - The role of nucleus accumbens and dorsolateral striatal D2 receptors in active avoidance conditioning. AB - The role of dopamine (DA) in rewarding motivated actions is well established but its role in learning how to avoid aversive events is still controversial. Here we tested the role of D2-like DA receptors in the nucleus accumbens (NAc) and the dorsolateral striatum (DLS) of rats in the learning and performance of conditioned avoidance responses (CAR). Adult male Wistar rats received systemic, intra-NAc or intra-DLS (pre- or post-training) administration of a D2-like receptor agonist (quinpirole) or antagonist ((-)sulpiride) and were given two sessions in the two-way active avoidance task. The main effects observed were: (i) sulpiride and lower (likely pre-synaptic) doses of quinpirole decreased the number of CARs and increased the number of escape failures; (ii) higher doses of quinpirole (likely post-synaptic) increased inter-trial crossings and failures; (iii) pre-training administration of sulpiride decreased the number of CARs in both training and test sessions when infused into the NAc, but this effect was observed only in the test session when it was infused into the DLS; (iv) post training administration of sulpiride decreased CARs in the test session when infused into the NAc but not DLS. These findings suggest that activation of D2 receptors in the NAc is critical for fast adaptation to responding to unconditioned and conditioned aversive stimuli while activation of these receptors in the DLS is needed for a slower learning of how to respond to the same stimuli based on previous experiences. PMID- 21619939 TI - Screening of species-specific lactic acid bacteria for veal calves multi-strain probiotic adjuncts. AB - The selection of promising specific species of lactic acid bacteria with potential probiotic characteristics is of particular interest in producing multi species-specific probiotic adjuncts in veal calves rearing. The aim of the present work was to select and evaluate in vitro the functional activity of lactic acid bacteria, Bifidobacterium longum and Bacillus coagulans strains isolated from veal calves in order to assess their potential use as multi species specific probiotics for veal calves. For this purpose, bacterial strains isolated from faeces collected from 40 healthy 50-day-calves, were identified by RiboPrinter and 16s rRNA gene sequence. The most frequent strains belonged to the species B. longum, Streptococcus bovis, Lactobacillus animalis and Streptococcus macedonicus. Among these, 7 strains were chosen for testing their probiotic characteristics in vitro. Three strains, namely L. animalis SB310, Lactobacillus paracasei subsp. paracasei SB137 and B. coagulans SB117 showed varying individual but promising capabilities to survive in the gastrointestinal tract, to adhere, to produce antimicrobial compounds. These three selected species-specific bacteria demonstrated in vitro, both singularly and mixed, the functional properties needed for their use as potential probiotics in veal calves. PMID- 21619940 TI - Mitochondrial plasmid-like elements in some hypovirulent strains of Cryphonectria parasitica. AB - In the chestnut blight fungus Cryphonectria parasitica, cytoplasmically transmissible hypovirulence phenotypes are elicited by debilitating mitochondrial DNA (mtDNA) mutations. In virus-free hypovirulent strains of C. parasitica from nature, the presence of a mitochondrial DNA element, named InC9, has been reported to cause similar disease syndromes. We have detected an additional mitochondrial element, termed plME-C9 (plasmid-like mitochondrial element C9) in some of the strains rendered hypovirulent by InC9. This element is a 1.4-kb DNA that exists in the mitochondria as monomeric and multimeric circular forms. Only a short 127-bp sequence of the plME-C9 DNA is derived from a region of the C. parasitica mtDNA that contains a reverse transcriptase-like open reading frame. The accumulation of the plME-C9 DNA in the mitochondria appears to adversely affect the growth of the fungus on synthetic medium. However, the presence plME C9 in different strains did not correlate with the manifestation of the hypovirulence phenotype, indicating that it is not the primary reason for the prevalence of attenuated C. parasitica strains in the Kellogg Forest in Michigan. PMID- 21619941 TI - Characterization of telomeres and telomerase expression in Xiphophorus. AB - Research investigating telomere lengths and telomerase expression in vertebrates has progressively become important due to the association of these two biological endpoints with cellular aging and cancer in humans. Studies that rely upon the traditional use of laboratory mice have been faced with limitations largely due to inbred mice possessing large telomeres and ubiquitous expression of telomerase. Recently, a number of small fish species have been shown to provide potentially informative models for examining the role of telomeres and telomerase within intact vertebrate animals. Xiphophorus fishes represent a new world live bearing genus that has not previously been assessed for telomere length or telomerase expression. To add to the knowledge base of telomere and telomerase biology in vertebrates we assessed telomere length and telomerase expression among several species of Xiphophorus. The telomere lengths in several organs (gill, brain, eyes, testis, ovary and liver) in three species (Xiphophorus hellerii, Xiphophorus maculatus, Xiphophorus couchianus) and also in F(1) interspecies hybrids were approximately 2-6 kb. This size was consistent within the same organs of the same species, as well as between species and F(1) hybrids. Despite possessing relatively short telomere lengths compared to humans, the consistency of size among Xiphophorus species and organs may allow experimental detection of telomere shortening. The relative expression of telomerase reverse transcriptase (TERT) was determined by quantitative real-time PCR. Expression levels of TERT was measured in seven organs (ovary, testis, liver, gill, brain, heart, skin) from X. maculatus, X. hellerii and in control and ultraviolet light (UVB) exposed skin samples from X. maculatus, X. hellerii, and F(1) interspecies hybrids. TERT gene expression was significantly higher in ovary and testis, while all other organs showed low relative TERT expression. Detectable increases in TERT expression were found in skin samples upon UVB exposure. Our findings suggest that Xiphophorus may serve as a suitable model for future studies investigating the association of telomere length and telomerase expression in regard to aging and disease. PMID- 21619942 TI - Chronic skin ulcer revealing metastasis from gastric cancer. PMID- 21619943 TI - Immune-mediated CNS diseases: a review on nosological classification and clinical features. AB - The immune-mediated diseases of the central nervous system (CNS) cover a wide range of clinical manifestations. Over the last years, considerable efforts have been made to establish a nosologic concept based upon distinctive pathophysiological characteristics of the single diseases. We describe the historically defined entities of immune-mediated diseases that primarily, but not exclusively, are affecting myelin structures. These include very rare entities as Schilder's, Balo's and Marburg's disease or the chronic and relapsing types of optic neuritis, for which evidence based paradigms still are virtually missing. In other, slightly more frequent diseases as neuromyelitis optica (NMO), advances in the concepts of specific biological features have been achieved and are beginning to transform into changes in clinical concepts. Acute disseminated encephalomyelitis (ADEM) and multiple sclerosis (MS) are by far the most frequent entities in this group and thus the only ones for which extensive empirical data on disease biology and evidence based clinical management strategies exist by now. For the most important entities, clinical features and therapeutic approaches are reviewed on the basis of current evidence. The results of basic science studies are assessed for their implications in nosological classification. PMID- 21619944 TI - Sudden sensorineural hearing loss: an autoimmune disease? AB - OBJECTIVES: To review our current knowledge of the pathogenesis of sudden sensorineural hearing loss, including viral infection, vascular occlusion and immune system-mediated mechanisms, and to discuss the pathogenesis as it relates to pharmacotherapy. SYSTEMATIC REVIEW METHODOLOGY: Relevant publications on the pathogenesis of sudden sensorineural hearing loss from 1944 to 2010 were analysed. RESULTS AND CONCLUSIONS: Sudden sensorineural hearing loss is defined as hearing loss of 30 dB in three sequential frequencies over 3 days or less. It can be an isolated symptom or the presenting symptom of a systemic disease. The aetiology and pathogenesis remain unknown. Detailed investigation typically reveals a specific cause in about 10% of patients. Proposed theories of causation include viral infections, vascular occlusion and immune system-mediated mechanisms. A variety of therapies have been proposed based on the various proposed aetiologies. PMID- 21619945 TI - Pathogenesis and therapies of immune-mediated myopathies. AB - The most common autoimmune muscle disorders include dermatomyositis (DM), polymyositis (PM), necrotizing autoimmune myositis (NAM) and sporadic inclusion body myositis (sIBM). DM is a complement-mediated microangiopathy leading to destruction of capillaries, hypoperfusion and inflammatory cell stress on the perifascicular regions. NAM is an increasingly recognized subacute myopathy triggered by statins, viral infections, cancer or autoimmunity with macrophages as the final effector cells causing fiber injury. PM and IBM are T cell-mediated disorders where cytotoxic CD8(+) T cells clonally expand in situ and invade major histocompatibility complex class I expressing muscle fibers. In sIBM, in addition to autoreactive T cells, there are degenerative features characterized by vacuolization and accumulation of stressor or amyloid-related misfolded proteins; an interrelationship between inflammatory and degeneration-associated molecules is prominent and enhances the cascade of pathogenic factors. These disorders are treatable, hence the need to make the correct diagnosis from the outset. The applied therapeutic strategies are outlined and the promising new agents are reviewed. PMID- 21619946 TI - Autoimmune diseases and rehabilitation. AB - This review gives an overview of the rehabilitation of autoimmune diseases. After general remarks on rehabilitation, the effects of acute and chronic exercises on inflammatory markers are summarized. Most of the available literature deals with rheumatoid arthritis (RA) and multiple sclerosis (MS), and therefore, rehabilitation of these diseases is described in more detail. Exercise is the main component in the rehabilitation of patients with RA and aims at increasing physical capacity, muscle strength, aerobic endurance, cardiovascular fitness and functional abilities, and helps to prevent secondary deconditioning due to reduced activity levels. Since MS causes a wide range of symptoms, the rehabilitation of these patients requires a multidisciplinary approach and encompasses physiotherapy, exercise therapy, hippotherapy, cognitive rehabilitation, psychological therapy, strategies to improve fatigue and coping programs. The ultimate goal of rehabilitation is to enable patients with chronic conditions to reach and maintain their optimal physical, sensory, intellectual, psychological and social functional levels, and to attain independence and self determination as far as possible. PMID- 21619947 TI - Autoimmune disorders affecting both the central and peripheral nervous system. AB - Various case series of patients with autoimmune demyelinating disease affecting both the central and peripheral nervous system (CNS and PNS), either sequentially or simultaneously, have been reported for decades, but their frequency is considerably lower than that of the "classical" neurological autoimmune diseases affecting only either CNS or PNS, such as multiple sclerosis (MS), chronic inflammatory demyelinating polyneuropathy (CIDP) or Guillain-Barre-Syndrome (GBS), and attempts to define or even recognize the former as a clinical entity have remained elusive. Frequently, demyelination started with CNS involvement with subsequent PNS pathology, in some cases with a relapsing-remitting course. Three potential mechanisms for the autoimmune etiology of these conditions can be discussed: (I) They could be caused by a common autoimmunological reactivity against myelin antigens or epitopes present in both the central and peripheral nervous system; (II) They could be due to a higher general susceptibility to autoimmune disease, which in some cases may have been caused or exacerbated by immunomodulatory treatment, e.g. b-interferon; (III) Their co-occurrence might be coincidental. Another example of an autoimmune disease variably involving the central or peripheral nervous system or both is the overlapping and continuous clinical spectrum of Fisher syndrome (FS), as a variant of GBS, and Bickerstaff brainstem encephalitis (BBE). Recent data from larger patient cohorts with demonstration of common autoantibodies, antecedent infections, and results of detailed clinical, neuroimaging and neurophysiological investigations suggest that these three conditions are not separate disorders, but rather form a continuous spectrum with variable central and peripheral nervous system involvement. We herein review clinical and paraclinical data and therapeutic options of these disorders and discuss potential underlying common vs. divergent immunopathogenic mechanisms. PMID- 21619948 TI - Genetic background of multiple sclerosis. AB - Multiple sclerosis (MS) is a one of the group of diseases labeled as "common complex". Virtually all common complex traits, genetic and environmental components have important roles, both independently and interactively, in disease susceptibility and stochastic and epigenetic effects cannot be overlooked. Data presented are largely part of the Canada-wide prospective, population-based longitudinal Canadian Collaborative Project on Genetic Susceptibility to MS (CCPGSMS) which includes over 30,000 unique families having at least 1 member with MS. Findings do not support a general propensity to autoimmune disease in MS families, but clearly highlight the importance of controlling for gender (patient, informant) when conducting such studies. The MHC class II association has been fine-mapped to the HLA-DRB5*0101-HLA-DRB1*1501-HLA-DQA1*0102-HLA DQB1*0602 extended haplotype. This HLA haplotype confers a relative risk of approximately 3 and homozygosity for this haplotype increases the risk by over 6 fold. However, the HLA haplotype loci interactions are complex and include, epistasis, trans and cis effects, and parent-of-origin effects. As well, there may be interactions of EBV and vitamin D with the HLA, In conclusion, using MS as an example, susceptibility for common complex disease most likely results from interactions of genes, environmental interactions and gene/environment interactions. PMID- 21619949 TI - Therapeutic blockade of TNF in patients with SLE-promising or crazy? AB - TNF is an important mediator of inflammation, but is also involved in the control of autoimmunity. The latter has been demonstrated in a murine model of SLE (NZB/W) and by the occurrence of autoantibodies to nuclear antigens as well as occasional, transient lupus-like syndromes in patients under TNF blockade. In contrast, data on increased TNF levels in serum, kidney and skin samples of SLE patients as well as results in other mouse models of the disease point to an inflammatory role of TNF in SLE organ disease. Despite all due caution, given these two sides of the cytokine, TNF blockade has by now been employed for several years in single cases and open label studies; data on more than fifty patients have meanwhile been published, for the vast majority of which infliximab was employed. These clinical data have to be very cautiously interpreted, as always with data on single cases or open label trials. However, some consistent pieces of information emerge and may inform controlled clinical trials: (i) While antibodies to double-stranded DNA commonly showed transient increases, lupus flares have not been seen so far and thus apparently are at least not the rule; (ii) in contrast, increases in anti-phospholipid antibodies may be associated with vascular adverse events; (iii) bacterial infections, pneumonia and urinary tract infections in particular, have been observed; (iv) short term induction therapy appears relatively safe, while long-term TNF blockade may confer significant risks in SLE; (v) TNF blocker induction therapy may lead to long-term remission in patients with lupus nephritis, hemophagocytic syndrome, and interstitial lung disease; (vi) patients with lupus arthritis often respond to TNF-blockade but symptoms recur after cessation of therapy, necessitating longer term therapy, which is more risky than short term treatment. PMID- 21619950 TI - Response to biotic and oxidative stress in Arabidopsis thaliana: analysis of variably phosphorylated proteins. AB - Protein phosphorylation plays a pivotal role in the regulation of many cellular events; increasing evidences indicate that this post-translational modification is involved in plant response to various abiotic and biotic stresses. Since phosphorylated proteins may be present at low abundance, enrichment methods are generally required for their analysis. We here describe the quantitative changes of phosphoproteins present in Arabidopsis thaliana leaves after challenging with elicitors or treatments mimicking biotic stresses, which stimulate basal resistance responses, or oxidative stress. Phosphoproteins from elicited and control plants were enriched by means of metal oxide affinity chromatography and resolved by 2D electrophoresis. A comparison of the resulting proteomic maps highlighted phosphoproteins showing quantitative variations induced by elicitor treatment; these components were identified by MALDI-TOF peptide mass fingerprinting and/or nanoLC-ESI-LIT-MS/MS experiments. In total, 97 differential spots, representing 75 unique candidate phosphoproteins, were characterized. They are representative of different protein functional groups, such as energy and carbon metabolism, response to oxidative and abiotic stresses, defense, protein synthesis, RNA processing and cell signaling. Ascertained protein phosphorylation found a positive confirmation in available Arabidopsis phosphoproteome database. The role of each identified phosphoprotein is here discussed in relation to plant defense mechanisms. Our results suggest a partial overlapping of the responses to different treatments, as well as a communication with key cellular functions by imposed stresses. PMID- 21619951 TI - Bisphosphonates do not alter the rate of secondary mineralization. AB - Bisphosphonates function to reduce bone turnover, which consequently increases the mean degree of tissue mineralization at an organ level. However, it is not clear if bisphosphonates alter the length of time required for an individual bone modeling unit (BMU) to fully mineralize. We have recently demonstrated that it takes ~350 days (d) for normal, untreated cortical bone to fully mineralize. The aim of this study was to determine the rate at which newly formed trabecular BMUs become fully mineralized in rabbits treated for up to 414 d with clinical doses of either risedronate (RIS) or alendronate (ALN). Thirty-six, 4-month old virgin female New Zealand white rabbits were allocated to RIS (n=12; 2.4 MUg/kg body weight), ALN (n=12; 2.4 MUg/kg body weight), or volume-matched saline controls (CON; n=12). Fluorochrome labels were administered at specific time intervals to quantify the rate and level of mineralization of trabecular bone from the femoral neck (FN) by Fourier transform infrared microspectroscopy (FTIRM). The organic (collagen) and inorganic (phosphate and carbonate) IR spectral characteristics of trabecular bone from undecalcified 4 micron thick tissue sections were quantified from fluorescently labels regions that had mineralized for 1, 8, 18, 35, 70, 105, 140, 210, 280, and 385 d (4 rabbits per time point and treatment group). All groups exhibited a rapid increase in mineralization over the first 18 days, the period of primary mineralization, with no significant differences between treatments. Mineralization continued to increase, at a slower rate up, to 385 days (secondary mineralization), and was not different among treatments. There were no significant differences between treatments for the rate of mineralization within an individual BMU; however, ALN and RIS both increased global tissue mineralization as demonstrated by areal bone mineral density from DXA. We conclude that increases in tissue mineralization that occur following a period of bisphosphonate treatment is a function of the suppressed rate of remodeling that allows for a greater number of BMUs to obtain a greater degree of mineralization. PMID- 21619952 TI - Obesity is a risk factor for fracture in children but is protective against fracture in adults: a paradox. AB - With the rise in obesity worldwide, an important debate has developed as to whether excess fat has a detrimental or protective effect on skeletal health in children and adults. Obese children appear to be over represented in fracture groups and recent evidence suggests that fat may be detrimental to bone accrual in children, although this effect may be confined to adolescence during rapid skeletal growth. Fat induced alterations in hormonal factors and cytokines during growth may play a pivotal role in disturbing bone accrual. In contrast, the widely accepted opinion is that fat appears to be protective of bone in adults and minimises bone loss in postmenopausal women. Recent evidence suggests that in adults, site specific fat depots may exert differing effects on bone (with visceral fat acting as a pathogenic fat depot and subcutaneous fat exerting protective effects), and that the effects of fat mass on bone and fracture risk may vary by skeletal site; obesity protects against hip and vertebral fractures but is a risk factor for fractures of the humerus and ankle. The incidence of fracture during adolescence is rising and osteoporosis remains a considerable health burden in older adults. Understanding the effects of fat mass on bone during growth and early adulthood is vital in informing future health strategies and pharmacotherapies to optimise peak bone mass and prevent fracture. PMID- 21619953 TI - Pulmonary langerhans cell histiocytosis with recurrent pneumothorax. AB - A 20-year-old woman with recurrent right pneumothorax was admitted to our hospital. A thoracoscopic bullectomy and lung biopsy was performed under general anesthesia. According to the histopathologic findings of permanent sections, a tissue specimen of diffuse lung disease revealed eosinophil infiltration and the presence of fibroblasts. Immunohistochemical stain showed S-100 protein-positive, as well as cluster of differentiation-68-positive large Langerhans cells. As a result, pulmonary Langerhans cell histiocytosis was diagnosed. The patient had an uncomplicated postoperative course and was discharged from the hospital on postoperative day 5. No recurrence was observed during the 6-month observation period. PMID- 21619954 TI - Successful treatment of atrioesophageal fistula by cervical esophageal ligation and decompression. AB - Atrioesophageal fistula is a rare yet devastating complication of transcatheter ablation for atrial fibrillation. This condition requires urgent intervention, but the optimal treatment strategy is yet to be defined. Reported therapies range from endoscopic stenting to direct atrial repair or reconstruction while on cardiopulmonary bypass. Here, we describe the successful management of an atrioesophageal fistula by cervical esophageal ligation and decompression, along with gastric drainage. PMID- 21619955 TI - Aortic root and right ventricular outflow tract reconstruction using composite biological valved conduits after failed Ross procedure. AB - The Ross procedure or pulmonary autograft has been frequently used for surgical treatment of aortic valve disease. One considerable disadvantage of the Ross procedure is the involvement of two valves (aortic and pulmonary) in treatment of single (aortic) valve disease. Both the aortic and pulmonary valves are at risk for future degeneration. Concurrent failure of both valves after a Ross procedure is rare but presents a significant technical challenge when reoperation is necessary. We describe a novel approach to this complication using composite biological valved conduits comprised of stentless bioprosthetic valves and polyester grafts. PMID- 21619956 TI - Aortic biological valve thrombosis in an HIV positive patient. AB - Biological aortic valve thrombosis is an exceptional complication. A 64-year-old patient positive for human immunodeficiency virus presented for syncope on exertion, 2 years after an aortic bioprosthetic valve replacement and double coronary artery bypass. Transvalvular aortic mean gradient was approximately 50 mm Hg on echocardiogram and catheterization. Cardiac computed tomography scan showed a limited opening of the bioprosthesis cusps. Surgical exploration revealed thrombosis of the three cusps on the aortic side, limiting the opening of the valve. No relation could be established between the patient's human immunodeficiency virus status and valve thrombosis. PMID- 21619957 TI - Culture of a prosthetic valve excised for streptococcal endocarditis positive for Aspergillus fumigatus 20 years after previous A fumigatus endocarditis. AB - Culture of a mitral valve prosthesis excised for streptococcal endocarditis yielded Aspergillus fumigatus. The patient had undergone valve replacement 20 years earlier for A fumigatus endocarditis. Data suggest that quiescent A fumigatus may have survived in a biofilm on the surface of the prosthesis. An antifungal therapy was initiated for 6 months. PMID- 21619958 TI - Resolution of an acute cardiac ischemia after the removal of a surgical drain in mitral and tricuspid valve repair. PMID- 21619959 TI - Primary cardiac extranodal B-cell lymphoma mimicking right atrial thrombus. PMID- 21619960 TI - Concomitant off-pump modified maze and coronary surgery. AB - We describe a modification of the Cox-Maze III using bipolar radiofrequency combined with off-pump coronary artery surgery for the treatment of patients with coronary artery disease and long-standing permanent atrial fibrillation. This study reports the midterm outcome of 12 patients with coronary artery disease and long-standing permanent atrial fibrillation who underwent off-pump coronary artery surgery and concomitant modified Maze with bipolar radiofrequency. At a mean follow-up of 23 months, all patients were alive, and 75% (9 of 12) had sinus rhythm. Our modified Maze can be safely and effectively combined with coronary artery surgery in an off-pump setting. PMID- 21619961 TI - Death and the cancer cell. PMID- 21619962 TI - The imperative for change: the STSA presidential address. PMID- 21619963 TI - Tumor necrosis as a prognostic factor for stage IA non-small cell lung cancer. AB - BACKGROUND: In stage IA non-small cell lung cancer (NSCLC), lobectomy and mediastinal lymph node dissection is considered the standard treatment. However, 20% to 30% of patients have cancer recurrences. The purpose of this study was to determine the patterns and risk factors for recurrence in patients with stage IA NSCLC. METHODS: We retrospectively reviewed the medical records of 201 patients who had confirmed stage IA NSCLC by lobectomy and complete lymph node dissection. RESULTS: There were 131 male patients with a mean age of 60.68+/-9.26 years. The median follow-up period was 41.4 months. Recurrences were reported in 16 patients. One hundred fourteen and 87 patients were T1a (<=2 cm) and T1b (>2 cm to <=3 cm), respectively. The pathologic results were as follows: adenocarcinomas and bronchioloalveolar carcinomas (n=134); squamous cell carcinomas (n=57); and other diagnoses (n=10). Tumor necrosis and lymphatic invasion were significant adverse risk factors for recurrence based on univariate analysis. Multivariate analysis showed that tumor necrosis was the only significant risk factor to predict cancer recurrence (hazard ratio, 4.336; p=0.032). The 5-year overall survival was 94.8% for necrosis-negative patients and 86.2% for necrosis-positive patients (p=0.04). The 5-year disease-free survival was 92.1% for necrosis negative patients and 78.9% for necrosis-positive patients (p=0.016). CONCLUSIONS: Tumor necrosis was shown to be an adverse risk factor for survival and recurrence in patients with stage IA NSCLC. Thus, close observation and individualized adjuvant therapy might be helpful for patients with stage IA NSCLC with tumor necrosis. PMID- 21619964 TI - Diffusion-weighted imaging is superior to positron emission tomography in the detection and nodal assessment of lung cancers. AB - BACKGROUND: Diffusion-weighted magnetic resonance imaging (DWI) makes it possible to detect malignant tumors based on the difference in the diffusion of water molecules among tissues. The aims of this study are to examine the usefulness of DWI compared with positron emission tomography-computed tomography (PET-CT) in the assessment of lung cancer, and the relationships between the apparent diffusion coefficient (ADC) value and several pathologic factors. METHODS: Sixty three patients with primary non-small cell lung cancer were enrolled in this study. The DWI and PET-CT were performed before surgery. There were 42 adenocarcinomas, 19 squamous cell carcinomas, and 2 other cell types. RESULTS: Sixty-one lung cancers (97%) were detected visually with DWI. This was significantly higher than 54 lung cancers (86%) with PET-CT. The accuracy for N staging by DWI was 0.81 (51 of 63), which was not significantly higher than 0.71 (45 of 63) by PET-CT. The sensitivity (0.75) for individual metastatic lymph node stations by DWI was significantly higher than that (0.48) by PET-CT. The specificity for individual nonmetastatic lymph node stations was 0.99 by DWI and 0.97 by PET-CT, respectively. The accuracy (0.95) for the diagnosis of lymph node stations by DWI was significantly higher than that (0.90) by PET-CT. There was a weak reverse relationship (correlation coefficient: 0.286) between the ADC value and the maximum standardized uptake value, but no relationship between ADC value and tumor size. The ADC values increased while the cell differentiation increased. CONCLUSIONS: Diffusion-weighted magnetic resonance imaging is superior to PET-CT in the detection of primary lesions and nodal assessment of non-small cell lung cancers. PMID- 21619965 TI - A prospective study to determine the incidence of non-imaged malignant pulmonary nodules in patients who undergo metastasectomy by thoracotomy with lung palpation. AB - BACKGROUND: To prospectively assess the incidence of non-imaged malignant nodules in patients who undergo thoracotomy for metastasectomy with bimanual lung palpation. METHODS: This is a prospective cohort study of patients who underwent open metastasectomy by thoracotomy. All patients had metastatic lung lesions, underwent 64-slice helical computed tomographic (CT) scan with intravenous contrast using 5-mm collimated cuts, and most had integrated PET (positron emission tomography)-CT. Unsuspected malignant pulmonary nodules that were palpated and removed, and that were not imaged preoperatively, were recorded. RESULTS: From January 2006 to March 2010, 152 patients underwent metastasectomy by rib-sparing, nerve-sparing thoracotomy by 1 surgeon. Fifty-one (34%) patients had 57 pulmonary nodules that were not imaged preoperatively and 32 of the 57 (56%) nodules were malignant. Thirty patients had non-imaged malignant nodules that were palpated and removed. There were 15 malignant nodules that were in different lobes than the imaged nodules. The 3 most commonly missed malignant nodules occurred in patients with colorectal cancer, renal cell, and sarcoma. CONCLUSIONS: Metastasectomy by thoracotomy, which affords bimanual palpation of the entire lung, discovers ipsilateral non-imaged malignant pulmonary metastases in 1 of 5 patients who had at least 1 imaged metastatic pulmonary lesion. This is true despite preoperative, fine cut chest CT scan with contrast, and integrated 18F-fluorodeoxyglucose-PET-CT scanning. The clinical significance of these non imaged, resected malignant nodules is unknown, nor is the added morbidity of resecting benign nodules. PMID- 21619966 TI - Forequarter amputation combined with chest wall resection: a single-center experience. AB - BACKGROUND: Forequarter amputation combined with chest wall resection is a rarely performed procedure. Six patients were treated for advanced malignancies with this operation in our institution since 1993. Uncontrollable pain, lymphedema, loss of function of the affected limb and, in some patients, localized ulceration of the tumor at the time of presentation, provided the indication for the operation. All patients underwent radical amputation of the upper limb and the structures of the shoulder girdle, in combination with resection of the thoracic chest wall in an extent of 2 to 7 ribs. METHODS: Chest wall reconstruction was achieved by implantation of a polytetrafluoroethylene patch (n=5) or a combination of a metal implant (Stratos System R, MedXpert GmbH, Heitersheim, Germany) and a polytetrafluoroethylene patch (n=1). Myocutaneous coverage of the defects was achieved by use of pedicled flaps from adjacent tissue (n=3) or by free myocutaneous flaps harvested from the amputated forearm (n=3). RESULTS: No perioperative mortality occurred; however, significant morbidity was seen after the use of the free forearm flaps based on occurring vascular problems. All 3 patients had to undergo surgical revision of the flap. Survival ranged from 5 to 50 months (median=23.5 months) with 3 patients still alive at the time of this investigation. CONCLUSIONS: Forequarter amputation in combination with chest wall resection is a feasible and potentially curative treatment for malignant tumors of the shoulder girdle with invasion of the chest wall. The operation results in immediate palliation and long-term survival can be obtained in selected cases. PMID- 21619967 TI - Invited commentary. PMID- 21619968 TI - Reoperations after failed transaxillary first rib resection to treat Paget Schroetter syndrome patients. AB - BACKGROUND: A series of 15 patients previously treated for Paget-Schroetter syndrome with a transaxillary first rib resection (TARR) were seen with recurrent thrombosis. METHODS: Ten were reoperated using an anterior subclavicular approach. The time of reoperation ranged from 5 months to 7 years (mean, 23.4 months). All patients had been subjected to multiple balloon plasties and 4 of them in addition had up to 3 stents implanted, which also failed. Reevaluation was done with venography. Ten patients were considered to be still salvageable and were reoperated, but 5 were inoperable due to progressive obliteration of the venous channel as early as 2 weeks after TARR. RESULTS: All 10 patients had successful reestablishment of the subclavian vein patency and caliber and have remained patent without anticoagulants. CONCLUSIONS: The patients who re thrombose or remain obstructed after TARR should be reoperated instead of resourcing to implanting stents or multiple balloon plasties that invariably fail, and patients should not be kept on anticoagulation indefinitely hoping to maintain the vein open. PMID- 21619969 TI - Invited commentary. PMID- 21619970 TI - Photodynamic therapy and the evolution of a lung-sparing surgical treatment for mesothelioma. AB - BACKGROUND: Photodynamic therapy (PDT) is a light-based cancer treatment that acts to a depth of several millimeters into tissue. This study reviewed the results of patients who underwent a macroscopic complete resection, by two different surgical techniques, and intraoperative PDT as a treatment for malignant pleural mesothelioma. METHODS: From 2004 to 2008, 28 patients with malignant pleural mesothelioma underwent macroscopic complete resection, 14 by modified extrapleural pneumonectomy (MEPP) and 14 by radical pleurectomy (RP) and intraoperative PDT. The surgical technique evolved over this period such that 13 of the last 16 patients underwent lung-sparing procedures, even in the setting of large-bulk tumors. RESULTS: Demographics in the MEPP and RP cohorts were similar in age, sex, stage, nodal status, histology, and adjuvant treatments. Stage III/IV disease was present in 12 of 14 patients (86%), with 50% or more with +N2 disease. The median overall survival for the MEPP group was 8.4 months, but has not yet been reached for the RP group at a median follow-up of 2.1 years. CONCLUSIONS: In addition to the inherent advantages of sparing the lung, RP plus PDT yielded a superior overall survival than MEPP plus PDT in this series. The overall survival for the RP plus PDT group was, for unclear reasons, superior to results reported in many surgical series, especially for a cohort with such advanced disease. Given these results, we believe RP plus PDT is a reasonable option for appropriate patients pursuing a surgical treatment for malignant pleural mesothelioma and that this procedure can serve as the backbone of surgically based multimodal treatments. PMID- 21619971 TI - Invited commentary. PMID- 21619972 TI - Meta-analysis of therapeutic procedures for acquired subglottic stenosis in adults. AB - BACKGROUND: Treatment for adult subglottic stenosis is technically demanding and no therapeutic algorithm exists. We performed the present meta-analysis of treatment for this condition in an attempt to compare efficacy on the basis of type of procedure. METHODS: We identified 24 eligible retrospective studies reporting the therapeutic results for inclusion criteria. Meta-analysis was performed by combining the results of the reported success rates; success is defined as a condition requiring no further treatment. The relative risk was used as a summary statistic. RESULTS: Pooled success rates of laryngotracheal resection and anastomosis (12 articles) and laryngoplasty with or without grafting (7 articles) were 95% and 76%, respectively, using a random-effects model. Success rates of endoscopic dilatation and laser resection (6 articles) varied between 40% and 82%. Meta-regression analysis showed a significant difference in the success rates between laryngotracheal reconstruction and laryngoplasty and between laryngotracheal reconstruction and an endoscopic procedure. When the indication for endoscopic management was a lesion size less than 1 cm, the results were significantly better. CONCLUSIONS: The success rate of laryngotracheal reconstruction is significantly higher than that of laryngoplasty or endoscopic intervention; however, endoscopic intervention is worth trying for lesions smaller than 1 cm without framework destruction. PMID- 21619973 TI - Use of bicaval dual-lumen catheter for adult venovenous extracorporeal membrane oxygenation. AB - BACKGROUND: Extracorporeal membrane oxygenation (ECMO) provides supplementary oxygenation and carbon dioxide removal for selected patients on mechanical ventilatory support. Venovenous ECMO is traditionally established by dual cannulation of the internal jugular and femoral veins. We report our institutional experience using single-site, dual-lumen cannula for venovenous ECMO as an alternative to the 2-catheter approach. This approach minimizes recirculation and avoids use of the femoral site, which confers potential advantages. METHODS: This is a retrospective review of a single institution's experience with a new bicaval dual lumen ECMO cannula. During a 19-month period, 27 consecutive patients were placed on ECMO using this catheter inserted through the right internal jugular vein. RESULTS: Single-site venovenous ECMO support was uneventfully initiated in 26 of the 27 patients (median age, 42 years; interquartile range, 31 to 58 years) and achieved full flows and adequate gas exchange. Median ventilator days before ECMO was 1 day (interquartile range, 0.25 to 3.5 days). The median duration of ECMO support was 9 days (interquartile range, 5.5 to 11.5 days). Decannulation was achieved in 70% of the patients and extubation in 59%. Two were bridged to lung transplantation and are still alive. The overall survival and hospital discharge rate was 56%. There was no device failure or in-cannula thrombosis. One superior vena cava injury occurred, and one cannula required repositioning. CONCLUSIONS: Single-site venovenous ECMO has advantages compared with traditional venovenous ECMO. Using image guidance, the cannula can reliably be used in prolonged venovenous ECMO cases. PMID- 21619974 TI - Morbidity of bleeding after cardiac surgery: is it blood transfusion, reoperation for bleeding, or both? AB - BACKGROUND: Etiology for increased morbidity in patients (2% to 8%) undergoing reoperation for bleeding after cardiac surgery is unclear. Recent work suggests that it may be related to red-cell transfusion, but what role does reoperation itself play? We sought to determine prevalence of and risk factors for reoperation for bleeding, separate the effect of reoperation from that of transfusion on hospital mortality and major morbidity, and identify the source of bleeding. METHODS: From January 1, 2000 to January 1, 2010, 18,891 primary and repeat coronary artery bypass grafting, valve, or combined operations were performed. Risk factors for reoperation were identified by multivariable logistic regression. Hospital mortality and major morbidity were compared in propensity matched patients requiring reoperation and not. Medical records from 2005 to 2010 were reviewed to determine bleeding source. RESULTS: A total of 566 patients (3.0%) underwent reoperation for bleeding, with considerable variability over time. Risk factors included older age, higher acuity, greater comorbidity, aortic valve surgery, longer myocardial ischemic and cardiopulmonary bypass durations, and surgeon. Mortality was higher for propensity-matched patients requiring reoperation; 8.5% (68% confidence interval [CI] 7.3% to 9.9%) versus 1.8% (CI 1.2% to 2.5%). Both greater transfusion and reoperation were independently associated with increased risk of mortality and major morbidity. At reoperation, technical factors (74%), coagulopathy (13%), both (10%), or other (3.3%) causes were responsible for bleeding. CONCLUSIONS: Transfusion and reoperation for bleeding both contribute to postoperative mortality and morbidity. Technical reasons are at the root of most bleeding, emphasizing a major focus for process improvement to minimize need for reoperation and blood use. PMID- 21619975 TI - Safety of same-day coronary angiography in patients undergoing elective aortic valve replacement. AB - BACKGROUND: Coronary artery imaging is required for most adult patients undergoing aortic valve replacement (AVR). METHODS: Between January 1, 2000, and December 31, 2007, 1413 patients underwent elective AVR at Mayo Clinic. Two propensity-matched groups at low risk for acute kidney injury were created: 321 patients with coronary angiography on the same day as AVR (same day) and 321 patients with coronary angiography more than 1 day before AVR (non-same day). We defined acute postoperative kidney injury with Acute Kidney Injury Network criteria (>=0.3 mg/dL or a 50% increase in serum creatinine from baseline). RESULTS: The same-day vs non-same-day groups were comparable in age (mean [SD] years: 70.6 [11.0] vs 70.8 [11.5]), sex (111 women [34.6%] vs 114 women [35.5%]), preoperative serum creatinine (1.14 [0.22] vs 1.15 [0.23] mg/dL), and ejection fraction (0.62 [0.12] vs 0.61 [0.12]). Coronary artery revascularization was performed in 118 patients (36.8%) in the same-day group and in 123 (38.3%) in the non-same-day group (p=0.68). Maximum (30-day) postoperative serum creatinine levels were not significantly different between the same-day and non-same day groups (1.30 [0.43] mg/dL vs 1.29 [0.42] mg/dL; p=0.87). Perioperative (30-day) acute kidney injury occurred in 75 patients (23.4%) in the same-day group and in 71 (22.3%) in the non-same-day group (p=0.99). Perioperative (30-day) death occurred in 5 patients (1.6%) in the same-day group and in 7 (2.2%) in the non same-day group (p=0.56). Other nonfatal complications rates were comparable between groups. CONCLUSIONS: In properly selected patients, coronary angiography can be performed the same day as elective AVR with no increase in perioperative morbidity or death. PMID- 21619976 TI - Invited commentary. PMID- 21619977 TI - Valve replacement for severe aortic stenosis with low transvalvular gradient and left ventricular ejection fraction exceeding 0.50. AB - BACKGROUND: Severe aortic stenosis with a low transvalvular gradient and preserved left ventricular ejection fraction (LVEF) is often misdiagnosed, leading to undertreatment of such patients with no clear indication for surgical intervention. This study investigated the outcome of aortic valve replacement (AVR) in patients with severe aortic stenosis and a low transvalvular gradient despite normal LVEF. METHODS: Between 1985 and 2008, we evaluated 73 patients who underwent AVR compared with 29 patients who did not. Overall, aortic valve area was 1.0 cm2 or smaller, LVEF was 0.50 or higher, and transvalvular gradient was 30 mm Hg or less. Multivariate and Cox analyses were used to compare these two groups according to AVR. RESULTS: Compared with controls, AVR patients were younger and with higher body mass index. Coronary artery bypass grafting was performed simultaneously in 38 AVR patients (52%). At follow-up (median, 42 months; interquartile range, 23 to 75 months), survival was longer in AVR patients. By Cox analysis, AVR remained a major predictor of lower mortality (hazard ratio, 0.237; 95% confidence interval, 0.119 to 0.470; p<0.0001). CONCLUSIONS: In patients with severe aortic stenosis and low transvalvular gradient despite a normal LVEF, AVR was associated with significant improvement in long-term survival and functional status and with a low operative mortality. PMID- 21619978 TI - Invited commentary. PMID- 21619979 TI - Invited commentary. PMID- 21619980 TI - Off-pump and on-pump coronary artery bypass grafting are associated with similar graft patency, myocardial ischemia, and freedom from reintervention: long-term follow-up of a randomized trial. AB - BACKGROUND: The Surgical Management of Arterial Revascularization Therapies trial was conceived to rigorously compare completeness of revascularization, clinical outcomes and resource utilization in unselected patients referred for elective, primary coronary artery bypass grafting randomly assigned to undergo off-pump (OPCAB) or conventional on-pump coronary artery bypass grafting using cardiopulmonary bypass (CPB). The goal of this follow-up study was to compare long-term survival, graft patency, myocardial ischemia, and clinical outcomes among survivors who volunteered to return for clinical evaluation and imaging studies. METHODS: Two hundred unselected patients with multivessel coronary artery disease were randomly assigned to OPCAB or CPB coronary artery bypass grafting between March 2000 and August 2001. All-cause mortality was determined by individual patient contact and referencing the Social Security Death Master File. Of 140 survivors, 87 volunteered to return after a minimum of 6.8 years (maximum, 8.4 years; mean, 7.5 years) for assessment of graft patency (computed tomographic angiography) and myocardial ischemia (cardiac positron emission tomography and 12-lead electrocardiogram). Age at follow-up ranged from 38 to 90 years (mean, 68 years). RESULTS: There were 26 deaths from all causes among OPCAB patients and 31 among CPB patients as of March 30, 2009. Graft patency was similar between groups among 622 grafts assessed by angiography before hospital discharge (99% OPCAB versus 97.7% CPB; p=0.22, Fisher's exact test), among 511 grafts assessed by angiography at 1 year (93.6% OPCAB versus 95.8% CPB; p=0.33), and among 190 grafts assessed by computed tomographic angiography at late follow up (76% OPCAB versus 83.5% CPB; p=0.44). Twelve of 34 OPCAB (35.3%) and 16 of 39 CPB patients (41.0%) had any ischemia on positron emission tomography scanning (p=0.62). Four OPCAB patients (11.8%) and 9 CPB patients (23.1%) had an ischemic region in excess of 10% of myocardium (p=0.21). At late follow-up, recurrent angina had occurred in 11 of 43 (25.6%) OPCAB patients and 5 of 44 (11.4%) CPB patients (p=0.09). Percutaneous reintervention had been performed at the discretion of blinded local cardiologists in 1 of 43 (2.3%) OPCAB patients and 1 of 44 (2.3%) CPB patients (p=1.0). No patient in either group has undergone repeat CABG. CONCLUSIONS: In this randomized trial, off-pump and on-pump coronary artery bypass grafting were associated with similar early and late graft patency, incidence of recurrent or residual myocardial ischemia, need for reintervention, and long-term survival. PMID- 21619981 TI - Invited commentary. PMID- 21619982 TI - Association of body mass index with new-onset atrial fibrillation after coronary artery bypass grafting operations. AB - BACKGROUND: Postoperative atrial fibrillation (AF) frequently complicates coronary artery bypass grafting (CABG) operations. As the frequency of obesity has increased in the United States, the number of obese patients undergoing CABG has kept pace. This study sought to define the association between body mass index (BMI) and postoperative AF. METHODS: We studied 12,367 consecutive patients with no history of AF who underwent isolated CABG operations. BMI was stratified according to Centers for Disease Control and Prevention criteria, and differences in baseline clinical and operative characteristics were adjusted through multivariate logistic regression models. RESULTS: The unadjusted incidence of new onset postoperative AF demonstrated a U-shape with regard to BMI. The highest incidence (34%) was found in the "lean" stratum (BMI<18.5 kg/m2), followed by 32% in the "severely obese" (BMI>=40 kg/m2) stratum. Lower incidences were found in the "normal" stratum (30%), in the "obese" stratum (28%), and the lowest incidence (26%) was in the overweight stratum. Observed incidence was 50% greater than the expected incidence in the "severely obese" stratum (32% vs 21%). In multivariate regression analysis adjusted for age and other covariates, BMI remains a strong risk factor for new-onset postoperative AF. Compared with normal BMI, obesity (odds ratio, 1.24; 95% confidence interval, 1.08 to 1.42) and severe obesity (odds ratio, 2.00; 95% confidence interval, 1.54 to 2.57) both emerged as strong risk factors for postoperative AF. No association was found between a lean BMI and postoperative AF (odds ratio, 1.14; 95% confidence interval, 0.66 to 1.98). CONCLUSIONS: After adjusting for potential confounders, obesity, as reflected by the body mass index, remains an independent predictor of postoperative AF. PMID- 21619983 TI - Invited commentary. PMID- 21619984 TI - Which troponometric best predicts midterm outcome after coronary artery bypass graft surgery? AB - BACKGROUND: Various troponin I measurements (troponometrics) have been used as surrogate markers of patient outcome after coronary artery bypass grafting (CABG). Our aim was to define the postoperative troponometric best able to predict in-hospital and late mortality. METHODS: In 440 patients (seen from January 2000 to September 2004) undergoing isolated on-pump CABG with standardized anesthesia, perfusion, cardioplegia, and postoperative care, we followed all-cause mortality (census June 2009, 100% complete). Subjects underwent troponin I (cardiac troponin I [cTnI]) estimation at baseline and 6, 12, 24, 48, and 72 hours postoperatively, and individual time-point cTnI (T6, T12, T24, T48, T72), peak cTnI (Cmax), increase in cTnI between 6 and 12 hours (T?6-12) and 6 and 24 hours (T?6-24), cumulative area under the curve cTnI (CAUC24, CAUC48, and CAUC72), and cTnI>=13 ng.mL(-1) at any time point were each analyzed using univariate and multivariable Cox models to identify the probability of in-hospital and late death. Logistic EuroSCOREs and calculated creatinine clearance (CrCl) were also included. The Akaike information criterion (AIC) was used to determine goodness of fit. RESULTS: There were 62 of 440 deaths after a median (interquartile range) follow-up period of 7.0 (5.7 to 8.1) years. Univariate Cox analysis demonstrated T12, T24, T48, T72, T?6-12, T?6-24, standardized CAUC24, CAUC48, and CAUC72 each to be predictors of midterm mortality. On Cox multivariable analysis in models incorporating both logistic EuroSCOREs and CrCl, both T72 (hazard ratio [HR], 95% confidence interval [CI], 1.10 [1.06 to 1.14]; p<0.001) and CAUC72 (1.45 [1.26 to 1.62], p<0.001) were identified as independent predictors of mortality. Of these, CAUC72 was superior based on the lowest AIC. CONCLUSIONS: In myocardial protection studies, serial troponin I data should be collected until 72 hours postoperatively to calculate CAUC72, as this troponometric best predicts midterm mortality. PMID- 21619985 TI - Antegrade cerebral perfusion with mild hypothermia for aortic arch replacement: single-center experience in 245 consecutive patients. AB - BACKGROUND: Aortic arch replacement remains a surgical challenge because of prolonged operative times, bleeding complications, and a considerable risk of neurologic morbidity and mortality. This study investigates our clinical results after modification of perfusion technique for cardiopulmonary bypass as well as temperature management for these high- risk patients. METHODS: Between January 2000 and January 2009, 245 consecutive patients underwent aortic arch repair during selective antegrade cerebral perfusion (ACP) with mild systemic hypothermia (30.5 degrees C+/-1.4 degrees C). Mean age was 63+/-12 years, 175 patients (71%) were men and 141 patients (58%) had acute type A dissection. Hemiarch replacement was performed in 152 patients (62%) while the remaining 93 patients (38%) underwent total arch replacement. RESULTS: Cardiopulmonary bypass time accounted for 168+/-62 minutes, and myocardial ischemic time was 103+/-45 minutes. Isolated ACP was performed for 38+/-27 (range 12 to 135) minutes. Chest tube drainage during the first 24 hours was 563+/-248 mL. Mean ventilation time was 44+/-22 hours. Serum lactate levels at 1, 12, and 24 hours postoperatively rose to 19+/-11, 33+/-14, and 20+/-8 mg/dL, respectively. We observed new postoperative permanent neurologic deficits in 14 patients (6%) and transient neurologic deficits in 12 patients (5%). The operative mortality rate was 8% (n=20). Among patients with ACP times 60 minutes or greater (n=28; 92+/-29 minutes), permanent neurologic deficits occurred in 2 individuals (n=2 of 28; 7%) and operative mortality was 7% (n=2 of 28). At late follow-up (3.8+/-3.2 years, 98% complete), 196 patients (80%) were still alive. CONCLUSIONS: Selective ACP in combination with mild hypothermia offered sufficient cerebral as well as distal organ protection in our patient cohort. Thus, current data suggest that this standardized perfusion and temperature management protocol can safely be applied to complex aortic arch surgery requiring up to 90 minutes of isolated ACP times. PMID- 21619986 TI - Invited commentary. PMID- 21619987 TI - Ablation of atrial fibrillation: comparison of catheter-based techniques and the Cox-Maze III operation. AB - BACKGROUND: Catheter-based ablation is often recommended for treatment of atrial fibrillation (AF), but there are no data that directly compare late results to those of the Cox-Maze procedure. Although catheter ablation avoids operation, lack of reliable transmurality may reduce effectiveness. We compared clinical outcomes of the cut-and-sew Cox-Maze procedure with catheter ablation. METHODS: Between January 1993 and October 2007, 97 patients aged 25 to 80 years underwent an isolated cut-and-sew Cox-Maze procedure. Patients were matched 1:2 according to age, sex, and AF type, with 194 patients undergoing catheter-based ablation for lone AF. RESULTS: At last follow-up, 82% of patients who underwent the Cox Maze procedure were free of AF and had stopped taking antiarrhythmic medications compared with 55% of patients who underwent ablation (p<0.001). When analyzed as a time-related event, freedom from recurrent AF was 87% 5 years after the Cox Maze procedure compared with 28% after catheter ablation (p<0.001). Late warfarin anticoagulation was required in 12% of patients who underwent the Cox-Maze procedure compared with 55% of patients who underwent ablation (p<0.001), and use of antiarrhythmic medications during follow-up was significantly higher in patients who underwent ablation (68% versus 15%, p<0.001). Forty-one patients (24%) required repeated ablation procedure and 9 required a second repeated ablation. CONCLUSIONS: Compared with catheter-based ablation, the Cox-Maze procedure results in greater freedom from AF and less medical treatment with antiarrhythmic drugs and warfarin anticoagulation during follow-up. PMID- 21619988 TI - Initial experience of sequential surgical epicardial-catheter endocardial ablation for persistent and long-standing persistent atrial fibrillation with long-term follow-up. AB - BACKGROUND: Patients with long-standing persistent (LSP) atrial fibrillation (AF) who have previously undergone catheter ablation represent a challenging patient population. Repeat catheter ablation in these patients is arduous and associated with a high failure rate, whereas surgical ablation can be complicated by multiple flutters. We sought to determine if minimally-invasive surgical ablation, followed by catheter ablation of all inducible flutters, would improve success rates over repeat catheter ablation alone. METHODS: Fifteen patients (Sequential) with persistent or LSP AF who failed at least one catheter ablation and one anti-arrhythmic drug (AAD) underwent surgical ablation, followed by planned endocardial evaluation and catheter mapping with ablation during the same hospitalization. Sequential patients were matched to 30 patients who had previously failed at least one catheter ablation and underwent a repeat catheter ablation (catheter-alone). The primary end point was event-free survival of any documented AF recurrence or AAD use. RESULTS: All patients underwent uncomplicated surgical ablation and electrophysiology procedure. Five Sequential patients had seven inducible flutters that were mapped and ablated. After a mean follow-up of 20.7+/-4.5 months, 13/15 (86.7%) Sequential patients, but only 16/30 (53.3%) catheter-alone patients, were free of any atrial arrhythmia and off of AAD (p=0.04). On AAD, 14/15 (93.3%) Sequential patients were free of any atrial arrhythmia recurrence, compared to 17/30 (56.7%) catheter-alone patients (p=0.01). CONCLUSIONS: For patients with atrial fibrillation who have failed catheter ablation, Sequential minimally invasive epicardial surgical ablation, followed by endocardial catheter-based ablation, has a higher early success rate than repeat catheter ablation alone. PMID- 21619989 TI - alphaB-crystallin improves murine cardiac function and attenuates apoptosis in human endothelial cells exposed to ischemia-reperfusion. AB - BACKGROUND: This study investigates the protective effect of exogenous alphaB crystallin (CryAB) on myocardial function after ischemia-reperfusion injury. METHODS: Mice underwent temporary left anterior descending artery occlusion for 30 minutes. Either CryAB (50 MUg) or phosphate-buffered saline (100 MUL [n=6, each group]) were injected in the intramyocardial medial and lateral perinfarct zone 15 minutes before reperfusion. Intraperitoneal injections were administered every other day. Left ventricular ejection fraction was evaluated on postoperative day 40 with magnetic resonance imaging. To investigate the effect of CryAB on apoptosis after hypoxia/reoxygenation in vitro, murine atrial cardiomyocytes (HL-1 cells) or human microvascular endothelial cells (HMEC-1) were incubated with either 50 MUg CryAB (500 MUg /10 mL) or phosphate-buffered saline in a hypoxia chamber for 6, 12, and 24 hours, followed by 30 minutes of reoxygenation at room air. Apoptosis was then assessed by western blot (Bcl-2, free bax, cleaved caspases-3, 9, PARP) and enzyme-linked immunosorbent assay analyses (cytoplasmic histone-associated DNA fragments and caspase-3 activity). RESULTS: On postoperative day 40, CryAB-treated mice had a 1.8-fold increase in left ventricular ejection fraction versus control mice (27%+/-6% versus 15%+/-4% SD, p<0.005). In vitro, (1) the HL-1 cells showed no significant difference in apoptotic protein expression, cytoplasmic histone-associated DNA fragments, or caspase-3 activity; (2) the HMEC-1 cells had increased but not significant apoptotic protein expression with, however, a significant decrease in cytoplasmic histone-associated DNA fragments (1.5-fold, p<0.01) and caspase-3 activity (2.7 fold, p<0.005). CONCLUSIONS: Exogenous CryAB administration significantly improves cardiac function after ischemia-reperfusion injury, in vivo. The protective anti-apoptotic affects of CryAB may target the endothelial cell. PMID- 21619990 TI - The myocardial protection of polarizing cardioplegia combined with delta-opioid receptor agonist in swine. AB - BACKGROUND: The purpose of this study was to determine whether polarized arrest using adenosine/lidocaine cold crystalloid cardioplegia in combination with the hibernation inductor delta-opioid receptor agonist pentazocine would give satisfactory myocardial protection rather than using depolarized supranormal potassium cardioplegia, supranormal potassium cardioplegia with pentazocine, or adenosine/lidocaine cardioplegia. METHODS: Twenty pigs were randomly divided into four groups (n=5 each) to receive the four types of cold crystalloid cardioplegia with an aortic cross-clamp time of 1 hour. Hemodynamic data were continuously measured, as was the left ventricular end-diastolic pressure (LVEDP), left ventricular end-systolic pressure (LVESP), plus or minus derivative of change in diastolic pressure over time (+/-dp/dt), cardiac output, pulmonary artery pressure, pulmonary capillary wedge pressure, cardiac troponin I, and left ventricular ultrastructure. RESULTS: Both the adenosine/lidocaine/pentazocine group and the adenosine/lidocaine group got significantly better results than the hyperkalemic and hyperkalemic pentazocine groups in improving hemodynamic values, pulmonary capillary wedge pressure, LVEDP, LVESP, +/-dp/dt, cardiac output, cardiac troponin I values, and left ventricular ultrastructure. There were no statistical differences between the adenosine/lidocaine/pentazocine group and the adenosine/lidocaine group at 1 hour after cross-clamp removal; but at 2 hours after cross-clamp removal, the adenosine/lidocaine/pentazocine group stands out (LVEDP 3.3+/-0.5, LVESP 122.5+/-18.9, +dp/dt 2.9+/-0.1, -dp/dt 2.0+/-0.6, cardiac output 2.6+/-0.4, and troponin I 4.9+/-0.5), with significant differences from the adenosine/lidocaine group (LVEDP 5.8+/-1.0, LVESP 98.5+/-10.1, +dp/dt 2.5+/ 0.2, -dp/dt 1.0+/-0.2, cardiac output 2.2+/-0.2, troponin I 8.2+/-0.8; p<0.05). The defibrillation rate was largely decreased after the cross-clamp was released in the group containing pentazocine in cardioplegia. CONCLUSIONS: Adenosine/lidocaine/pentazocine cold crystalloid cardioplegia gave satisfactory cardiac arrest and better myocardial protection than the other three groups, especially with regard to improving prolonged postoperative cardiac function. PMID- 21619991 TI - Hybrid video-assisted thoracic surgery-robotic minimally invasive right upper lobe sleeve lobectomy. AB - PURPOSE: Video-assisted thoracoscopic (VATS) lobectomy has been demonstrated to be safe and feasible. However, only a few reports exist on minimally invasive sleeve lobectomy. In most of these, bronchial anastomoses were accomplished in an open surgical technique through a minithoracotomy. We report on a combined robotic and VATS approach for a true minimally invasive right upper sleeve lobectomy. To our knowledge it is the first report of this kind. DESCRIPTION: A 30-year-old female patient presented with a low grade neuroendocrine tumor occluding the orifice of the right upper lobe bronchus. A complete minimally invasive right upper sleeve lobectomy was performed. Dissection and individual control of the right upper lobe pulmonary vessels was performed through three 1.5 to 2 cm incisions by means of conventional VATS. The specimen was retrieved through the upper thoracocentesis, which was enlarged to 4.5 cm. For airway reconstruction, a bronchial anastomosis between the right intermediate and the right main bronchus was performed with the da Vinci robot through the same incisions. EVALUATION: Intraoperative blood loss was minimal and total operative time was 364 minutes. The postoperative course was uneventful. Chest X-rays showed no sign of atelectasis. The chest tube was removed on postoperative day 9 due to prolonged pleural effusion. Bronchoscopy on postoperative day 14 showed a patent right airway and a well healing anastomosis. The patient was discharged on postoperative day 15. CONCLUSIONS: The da Vinci robot provides a benefit in more complex thoracic procedures like pulmonary sleeve resections. It is an ideal tool to perform delicate surgical maneuvers in vulnerable and difficult to reach anatomic areas. The study of the literature shows that VATS sleeve lobectomy is being performed in dedicated centers only with a low morbidity. PMID- 21619992 TI - Cystic degeneration of a carcinoid lung tumor: an uncommon cause of an intrathoracic cyst. AB - Carcinoid tumor is a rare, slow-growing neuroendocrine tumor that accounts for less than 1% of all lung tumors. It occurs most commonly in the midgut, then the lung, and usually presents as a solid lesion. We report a patient with a typical carcinoid tumor that had undergone cystic degeneration. PMID- 21619993 TI - Mediastinal coelomic cyst. AB - Coelomic cysts are rare cysts of mesothelial origin. This is a case report of a thoracic coelomic cyst presenting as atypical chest pain. PMID- 21619994 TI - Empyema thoracis complicated by pneumothorax necessitans manifesting as lobulated, localized subcutaneous emphysematous swellings. AB - We describe a patient with empyema thoracis that had eroded through the apical chest wall to the supraclavicular fossa and lung parenchyma, ultimately leading to a bronchopleural fistula manifesting as lobulated, localized subcutaneous emphysematous swellings over the chest wall and supraclavicular fossa. This is a rarely reported phenomenon and the reason why the lobulated subcutaneous emphysematous swellings remained localized is unknown. PMID- 21619995 TI - Mature cystic teratoma with malignant transformation to adenocarcinoma. AB - An anterior mediastinal tumor had been found in a 65-year-old woman 10 years previously, and she had been followed-up under the diagnosis of a mature cystic teratoma. Changes in the structure of the inside of the mediastinal cystic tumor were observed on a chest computed tomography image, and she was referred to our hospital for surgical intervention. The tumor was removed and was determined to be a mature teratoma. An adenocarcinoma was also observed in part of this cystic lesion. From these findings, the lesion was diagnosed as a malignant change in a mediastinal mature teratoma. Because few if any changes tend to be observed in the cystic wall of such cases, an aggressive resection of such mature cystic teratomas is therefore recommended. PMID- 21619996 TI - Multiple thymic carcinoids. AB - We present a very rare case of simultaneous thymic carcinoids with similar size, one of which showed spontaneous regression. A 68-year-old man was admitted to the hospital because of two similar abnormal masses at his anterior mediastinum on chest computed tomography, one of which had decreased from 25 to 16 mm in diameter. A total thymectomy was performed and the pathologic examinations revealed that both tumors were atypical carcinoids. There have been seldom reports of multiple thymic carcinoids, and this case might suggest that total thymectomy is the best way to treat thymic carcinoid because of the possibility of multicentric origins. PMID- 21619997 TI - Thoracic outlet syndrome after the Nuss procedure for the correction of extreme pectus excavatum. AB - Since the Nuss procedure was introduced in 1998, many complications have been reported, but not thoracic outlet syndrome. Here we report a 13-year-old boy with pectus excavatum who had thoracic outlet syndrome develop after a modified Nuss procedure. The major modification from the original technique was the use of an additional bar to resolve his long, asymmetric deformity. The patient showed clinical features of brachial plexus compression. The abrupt structural and spatial changes induced by the marked elevation of the upper depressed chest might have given rise to this condition. Thoracic outlet syndrome is a possible complication of the Nuss procedure. PMID- 21619998 TI - Platelet transfusion associated with acute lung injury after coronary artery bypass grafting. AB - Transfusion-related acute lung injury is a potentially fatal complication of blood and plasma transfusion; however, the incidence relating to platelet use in cardiac surgery is uncommon. In the presence of normal left ventricular function, an acute increase in pulmonary capillary permeability leads to a high protein content pulmonary edema, which leads to a dramatic reduction in pulmonary function due to acute lung injury and also intravascular fluid depletion. The clinical picture is acute and the condition is associated with considerable mortality. Although the exact mechanism of transfusion-related acute lung injury is unknown, it may be due to an antibody-mediated reaction caused by preformed leukocyte antibodies or activation of inflammatory mediators. The signs, diagnosis, and therapeutic interventions are discussed with reference to a case report. PMID- 21619999 TI - Successful combined minimally invasive direct coronary artery bypass and transapical aortic valve implantation. AB - Transapical aortic valve implantation is indicated in high-risk patients with aortic stenosis and peripheral vascular disease requiring aortic valve replacement. Minimally invasive direct coronary artery bypass grafting is also a valid, minimally invasive option for myocardial revascularization in patients with critical stenosis on the anterior descending coronary artery. Both procedures are performed through a left minithoracotomy, without cardiopulmonary bypass, aortic cross-clamping, and cardioplegic arrest. We describe a successful combined transapical aortic valve implantation and minimally invasive direct coronary bypass in a high-risk patient with left anterior descending coronary artery occlusion and severe aortic valve stenosis. PMID- 21620000 TI - Balloon-assisted deployment prevents endograft misalignment during hybrid aortic arch repair. AB - Misaligned deployment refers to eccentric flaring of the bare stent portion of the endograft that causes proximal device retroflexion resulting in suboptimal graft placement. A 71-year-old woman with 6.8-cm distal aortic arch aneurysm underwent hybrid repair with combination of open aortic arch debranching and subsequent thoracic endovascular aortic repair. During endograft deployment, misaligned opening occurred; this was corrected by using a balloon-assisted graft deployment. Follow up imaging continues to demonstrate complete aneurysm exclusion. Misaligned deployment is a clinical challenge; we hereby present a previously undescribed technique to accommodate this limitation of the current endograft technology. PMID- 21620001 TI - Mitral regurgitation caused by an isolated mitral leaflet cleft. AB - An isolated cleft of the mitral valve leaflet is rare cause of mitral regurgitation in adults. We report a successful minimally invasive mitral valve repair for severe mitral regurgitation caused by an isolated cleft of the anterior mitral leaflet. During the operation, we found a large cleft measuring 5*8 mm in the center of the anterior mitral leaflet. We closed the cleft directly and performed annuloplasty with a 30-mm Carpenter-Edwards Physio Ring (Edwards Lifesciences, Irvine, CA). The mitral valve is very well visualized with the video-assisted minimally invasive approach through the right chest. PMID- 21620002 TI - Occlusion of left coronary artery ostium by left coronary cusp fusion. AB - We report two cases of adults who had occlusion of the left coronary artery ostium by the left coronary cusp. Both patients were treated with the Bentall procedure, and both revealed a good, long-term course. We offer proper strategy of perioperative management consisting of precise preoperative diagnosis with a multimodality imaging study and adequate operative technique. PMID- 21620003 TI - Closed chest resection of left ventricular myxoma through thoracoscopic assistance. AB - We performed a totally endoscopic resection of a left ventricular myxoma using the Heart Port Endoclamp System and conventional endoscopic instruments in a young male patient. It is a feasible and safe procedure with good clinical results and an excellent cosmetic outcome. PMID- 21620004 TI - Endocarditis of bovine jugular vein conduit due to Q fever. AB - Contegra (Medtronic, Minneapolis, MN) conduits are routinely used in cases of right ventricular outflow tract reconstruction during congenital heart surgery. We report two cases of Q fever endocarditis involving Contegra conduits. Surgical treatment and distinct aspects of both unusual cases are described. PMID- 21620005 TI - Capillary hemangioma mimicking a dumbbell-shaped mediastinal neurinoma. PMID- 21620006 TI - Dramatic postchemotherapy reduction of left atrial involvement from lung cancer. PMID- 21620007 TI - Left innominate venous aneurysm presenting as an anterior mediastinal mass. PMID- 21620008 TI - Extracorporeal membranous oxygenation and left atrial decompression: a fast and minimally invasive approach. AB - Left atrial decompression for patients under extracorporeal membranous oxygenation is sometimes mandatory, but it may be technically difficult. We describe a safe and minimally invasive technique to perform this while placing a small cannula antegradely in the pulmonary artery trunk. PMID- 21620009 TI - Double-barreled cannon stent grafts: possible solution for extremely dilated landing zone of aorta. AB - Anatomical limitations have been identified as barriers to the more widespread application of thoracic endovascular aortic repair. Here, we report a case in which a novel technique was used as a solution for an extremely dilated aortic landing zone. An elderly gentleman in profound shock was diagnosed with a ruptured distal aortic arch aneurysm. Open repair was considered, but owing to the presence of multiple comorbidities and an extremely dilated aorta in the landing zone (45 mm) which made thoracic endovascular aortic repair infeasible, it appeared to entail a high risk for perioperative morbidity and mortality. Therefore, to save the patient's life, we designed a novel double-barreled cannon thoracic endovascular aortic repair method, and further developed a formula for choosing appropriate stent graft sizes. The patient gradually recovered to baseline physical status after the operation. PMID- 21620010 TI - Technique for insertion of HeartMate II left ventricular assist device inflow cannula. AB - During implantation of the HeartMate II left ventricular assist device, the inflow cannula is typically inserted in such a way that the sewing ring is placed within the ventricular cavity in contact with the endocardium. We describe a novel implantation technique in which the sewing ring is secured to the left ventricular epicardium, thus avoiding the complication of bleeding from the exposed cut surface of apical myocardium. In addition, the shortened intraventricular segment of the inflow cannula protects against inflow occlusion by the interventricular septum. PMID- 21620011 TI - Modified T-shaped left atrium incision in the bi-atrial approach for infracardiac type TAPVR repair. AB - The bi-atrial approach "in-situ" technique of vertical confluence to the left atrial repair avoids the risks of anastomosis site kinking and twisting for infracardiac-type total anomalous pulmonary venous connection reconstruction, but how to obtain the adequate left atrial orifice size is an important issue. Herein, we describe our vertical confluence vein flap with the modified T-shaped left atrium incision technique in managing those patients with infracardiac type total anomalous primary venous reconstruction. PMID- 21620012 TI - Surgical management of endocarditis: the society of thoracic surgeons clinical practice guideline. PMID- 21620013 TI - Single incision video-assisted thoracic surgery using a laparoscopic port. PMID- 21620015 TI - Factors associated with the development of acute heart failure in critically ill patients with severe pandemic 2009 influenza A (H1N1) infection. PMID- 21620017 TI - Spare the blood, but save the kidneys. PMID- 21620019 TI - Impact of postoperative atrial fibrillation after CABG. PMID- 21620021 TI - Right ventricular outflow tract reconstruction: the impact of allograft characteristics. PMID- 21620023 TI - Patients with interrupted aortic arches and small aortic annulus: a rare entity? PMID- 21620024 TI - Preface: 34th Congress of the Italian Transplantation Society (SITO). PMID- 21620025 TI - Results with expanded donor acceptance criteria in heart transplantation. AB - OBJECTIVE: Over the past years both donor and recipient profiles have changed in heart transplantation. Satisfactory clinical outcomes of marginal donors in candidates >60 years of age have led us to allocate suboptimal donors to younger recipients as well. Therefore, we retrospectively reviewed our experience. METHODS: Among 199 patients undergoing heart transplantation from January 2000 to February 2010, there were 83 (41%) aged 61-72 years. The other 116 (59%) ranged in age between 18 and 60 years. According to their clinical conditions as heart transplantation candidates, They were classified into 4 groups: younger recipients (n=116) of either optimal donors (n=72; group 1 [G1]) or marginal donors (n=44; group 2 [G2]) and older recipients (n=83) of either marginal grafts (n=70, group 3 [G3]) or optimal grafts (n=13; group 4 [G4]). The gender distribution, cause of end-stage heart failure, preoperative pulmonary hypertension incidence, pretransplantation clinical status, and mean follow-up were not significantly different among the 4 groups. RESULTS: Overall 30-day survival was 90 +/- 1% and 10-year rate was 78 +/- 9%. Among the groups, 30-day and 10-year actuarial survival rates were, respectively: 94 +/- 4% and 87 +/- 1% for G1; 86 +/- 5% and 84 +/- 7% for G2; 88 +/- 4% and 71 +/- 7% for G3 and were 100% and 82 +/- 7% for G4 (P=.7). In comparison among the 4 groups, there was no significant difference regarding freedom from graft failure (P=.3), right ventricular failure (P=.3), acute rejection episodes (P = .2), chronic rejection (P=.2), neoplasia (P=.5), or chronic renal failure (P=.1). Older recipients of marginal donors [G3] had a 4% (n=3) prevalence of permanent pacemaker implant, versus G2: 3% (n=2) among (P=.1). CONCLUSION: Our results suggest that extended donor and recipient criteria do not compromise clinical outcomes after transplantation. PMID- 21620026 TI - Magnetic resonance with diffusion-weighted imaging in the evaluation of transplanted kidneys: preliminary findings. AB - BACKGROUND: The aim of this study was to compare values of apparent diffusion coefficient (ADC) and diffusion (D) with renal function indexes, in a population of kidney transplant recipients who underwent magnetic resonance with diffusion weighted imaging (DWI) of the transplanted graft. MATERIALS AND METHODS: We studied 21 patients using a 1.5-Tesla magnetic resonance; DWI sequences were acquired with several b-values. Patients were divided into 3 groups by their creatinine clearance values: group A, clearance >60 mL/min; group B, clearance >30 and <=60 mL/min; and group C, clearance <=30 mL/min. ADCs values between groups were compared using the Mann-Whitney U test. Receiver operating characteristic (ROC) curves were created for prediction of normal renal function (group A) and renal failure (group C). RESULTS: Comparing mean values of ADC between groups A and C, we observed a difference (P=0.0012), with higher values in group A. Regarding mean values of D, we observed a difference between groups A and C (P=0.022). In the comparison between contiguous groups, we observed no difference for ADC and D values. In the prediction of normal clearance values (group A), ROC curve showed an area under curve (AUC) of 0.861, with a sensitivity of 88.89% and specificity of 75% using a threshold ADC value >=2.1 * 10(-3) mm(2)/sec. For prediction of normal clearance values (group A), ROC curve showed an AUC of 0.787, with a sensitivity of 77.8% and specificity of 83.3% using a threshold D value >=2.3 * 10(-3) mm(2)/sec. CONCLUSION: Although studies with a larger number of patients are needed, DWI represents a promising tool for noninvasive assessment of renal function. An ADC >= 2.1 * 10(-3) mm(2)/sec and a D >= 2.3 * 10(-3) mm(2)/sec may be used as a threshold for predicting normal clearance. PMID- 21620027 TI - Kidney transplantation from hepatitis B virus core antibody-positive donors: prophylaxis with hepatitis B immunoglobulin. AB - OBJECTIVE: Hepatitis B virus core antibody (HBcAb)-positive organ donors have the potential to transmit infection to transplant recipients. PATIENTS AND METHODS: We investigated the use of a single dose of 2000 IU of hepatitis B immunoglobulin in 18 patients among a population of 54 kidney transplant recipients from HBcAb positive deceased donors. RESULTS: Twelve recipients were HBcAb-positive before transplantation. Among the other 42 patients, 5 (11.9%) seroconverted from HBcAb negative to HBcAb-positive, whereas one HBcAb-positive recipient became hepatitis B virus surface antigen-positive with clinical signs of active hepatitis 6 years after transplantation. In the 18 patients who underwent prophylaxis, we did not find any seroconversion or hepatitis B virus (HBV) transmission. Graft and patient survival of HBcAb-positive kidney transplants did not differ significantly with a matched population of HBcAb-negative transplantation. CONCLUSION: These results suggest that kidney transplantation from HBcAb-positive donors is safe with a low rate of HBV transmission. A prophylaxis with a single shot of hepatitis B immunoglobulin may be effective in reducing the risk of HBV seroconversion or reactivation and may be suggested in all naive or HBcAb positive transplant recipients. PMID- 21620028 TI - Extended criteria liver donation and transplant recipient consent: the European experience. AB - The organization known as ELPAT (Ethical, Legal and Psychological Aspects of Organ Transplantation) coordinated the distribution of an electronic questionnaire concerning the definition of extended criteria liver donation (ECD) and the implication for informed consent of transplant recipients to European liver transplant centers. Completed questionnaires were received from 30 centers in 13 countries. Twenty-eight centers accepted ECD liver donors. The criteria for defining a liver donor as ECD were: steatosis in 24 centers (85%); age up to 80 years in 23 centers (82%); serum sodium levels higher than 165 mmol/L in 17 centers (60%); intensive care unit stay with ventilation longer than 7 days in 16 centers (57%); serum glutamic oxalo-acitic transaminase levels higher than 90 U/L in 12 centers (42%); body mass indeces more than 30 in 10 centers (35%); serum glutamic pyruvic transaminase levels higher than 105 U/L in 10 centers (35%); serum bilirubin levels higher than 3 mg/dL in 10 centers (35%); and other criteria in 13 centers (46%). Twenty-three centers informed the transplant candidate of the ECD status of the donor: 10 centers (43%) when the patient registered for transplantation, 3 centers (14%) when an ECD liver became available, and 10 centers (43%) on both occasions. Ten centers required the liver transplant candidate to sign a special consent form. Ten centers informed the potential recipient of the donor's serology. Only three centers informed the potential recipient of any high risk behavior of the donor. PMID- 21620029 TI - Donor-Model for End-Stage Liver Disease and donor-recipient matching in liver transplantation. AB - BACKGROUND: The product between donor (D) age and recipient (R) Model for End Stage Liver Disease (MELD) score at the moment of liver transplantation (LT) has been proposed as a potential D-R matching tool to reduce the risk of "futile" LT from using the MELD score as the main allocation tool. The aim of this study was to evaluate the prognostic ability of D-MELD among a cohort of Italian patients already selected for LT on the basis of a D-R matching philosophy. METHODS: We studied 303 consecutive adult patients undergoing first LT for chronic liver diseases with available D-MELD at the moment of LT from 2003 to 2009. Optimal donors were assigned to more severe cirrhotic patients (MELD >=20); suboptimal organs were allocated to patients with hepatocellular carcinoma (HCC) not responsive to bridging therapies (specific priority score) or other exceptions with MELD <20. A suboptimal donor had age >70 years, severe steatosis by ultrasound, and/or body mass index >30 kg/m(2), partial liver, or hepatitis C (HCV) or B virus positivity. RESULTS: Characteristics of the study group were a median age of 55 years (range, 27-68 years), HCV positivity in 164 patients (54%), HCC in 134 patients (44%), partial liver use in 25 (8%), MELD 15 (range, 6 40), D-age of 56 years (range, 18-87 years), and median D-MELD score 826 (range, 126-2,988). Overall graft survival was 84%, 79%, and 77% at 1, 3, and 5 years after LT, respectively. Logistic regression did not show a significant correlation between graft failure and D-MELD score in the absence of a significant D-MELD cutoff. Cox regression with D-MELD as the continuous variable showed a hazard ratio (HR) of 0.99 (95% confidence interval [CI], 0.99-1.00; P=NS); and with D-MELD as a dichotomic variable (>=0 to <1,600) an HR of 0.98 (95% CI, 0.63-1.77; P=NS). CONCLUSION: The prognostic ability of D-MELD fails in OLT centers that use a more complex D-R matching policy. PMID- 21620030 TI - Italian continuing medical education program: an evidence-based course for surgeons and health personnel of a regional transplantation center. AB - In the organ transplantation area the application of the evidence-based medicine (EBM) methods may be limited by several, heterogeneous conditions, eg, mandatory laws and protocols, logistic concerns, as well as donor/recipient matching. In this report we have described the results of a first EBM-oriented course for surgeons and health personnel in a regional transplantation center under the Italian Continuing Medical Education (CME) program. The course was formally approved for 25 credits. It included a maximum of 70 medical and nursing staff registrants; 50 of the spots were reserved for our transplantation center. The course was scheduled in 10 sessions from June to November 2010. Each session was composed of 2 phases: (1) first, computer-assisted education including slide presentations (2 hours); and (2) subsequent discussion led by experts (1.5 hours). The registered participants were expected to be able to correctly answer a multiple-choice, 10-question questionnaire at the end of each session. The majority of the participants considered the course relevant to their need to be updated and effective to improve their clinical skills. The requirements to obtain credits by the Italian CME program for live events were overall presence >=80% and correct responses in the postsession questionnaires >=70%. However, among the initially registered participants 31.5% failed at least one of these requirements. The main reason for failure was exceeding the maximum number of absences. Paradoxically, the absences were largely caused by the simultaneous execution of surgical/medical transplantation procedures. For professional figures engaged in complex medical activities, the Italian CME program should consider different threshold limits for the maximum number of absences allowed at live events. PMID- 21620031 TI - Analysis of the motivation for hematopoietic stem cell donation. AB - The Italian Bone Marrow Donor Register is the institutional organization for management of unrelated hematopoietic stem cell donors. The law requires only a donor's clinical history, but not a psychosocial profile for registration. We have studied the donor's motivation for enlistment on the donor registry and the medical staff's need for this information to interact correctly with the donor. For this purpose we distributed a questionnaire to new donors at the 20 centers in the Lombardy Region over a period of 1 year. The analysis of the responses revealed a prevalence of extrinsic motivations that would not ensure continued registration for donation. Therefore, it is necessary that the donor be well informed and better educated about all aspects of donation, in order to produce a shift to an intrinsic motivation. This objective can be facilitated via professional training of health workers in communication. PMID- 21620032 TI - Continuous infusion of local anesthesia after living donor nephrectomy: a comparative analysis. AB - INTRODUCTION: Today local anesthetic wound infiltration is widely recognized as a useful adjunct in a multimodality approach to postoperative pain management. The effectiveness of continuous wound infusion of ropivacaine for postoperative pain relief after laparoscopic living donor nephrectomy was analyzed in this retrospective, comparative analysis. METHODS: Twenty patients undergoing living donor nephrectomy were divided into two groups: standard analgesic therapy (n=10) and ropivacaine continuous infusion group (n = 10). RESULTS: We observed a significant difference in term of visual analogue scale scores, use of morphine, hospital stay, and bowel recovery in favor of the ropivacaine group. The cost analysis demonstrated an overall savings of 985 Euros/patient. DISCUSSION: Surgical wound infusion with ropivacaine was safe and seemed to improve pain relief and accelerate recovery and discharge, reducing the overall costs of care. Postoperative pain control in the donor is of primary importance for better patient compliance and greater perceived quality of health care service. PMID- 21620033 TI - Ethical issues with nondirected ("good samaritan") kidney donation for transplantation. AB - "Good samaritan" donation has been of great interest in Italy. At the request of the Presidency of the Council of Ministers, the National Committee on Bioethics expressed its opinion on the matter. While highlighting its controversial aspects, the assessment was favorable. The National Council for Health established working criteria. Yet eminent bioethicists sharing the same values have reached discordant conclusions. Legal developments leading to the authorization of living donor kidney transplants from blood relatives or emotionally close individuals may offer a path for ethical assessment of the practice. PMID- 21620034 TI - The consumption of glucose during ex vivo lung perfusion correlates with lung edema. AB - INTRODUCTION: Ex vivo lung perfusion (EVLP) has been recently proposed to recondition organs before transplantation from donors with marginal or unacceptable features. The aim of our investigation was to explore glucose consumption during EVLP. MATERIALS AND METHODS: We investigated 8 domestic pigs (mean weight, 21 +/- 0.8 kg). After perfusion with Perfadex, retrieval, and back table surgery, we initiated EVLP. The lungs were perfused with Steen solution with added methylprednisolone, cefazoline, and heparin. The blood flow was gradually increased with a target of 40% of the estimated cardiac output (or less if the pulmonary artery pressure was >15 mm Hg), while keeping the left atrial pressure between 3 and 5 mm Hg. The temperature of the perfusate was increased from 25 degrees C to 37 degrees C. Once the temperature of the lung outflow was >32 degrees C, we began gas flow (4 L/min, 5%-8% CO(2) in air) and mechanical ventilation. EVLP parameters and blood gases were measured throughout the experiment; glucose consumption was calculated as (glucose initial-glucose final)/time. The wet to dry ratio was also calculated as an index of lung edema. RESULTS: When stratified by median glucose consumption (0.237 mg/min), high glucose consumers (0.588 +/- 0.17) were characterized by worse lung function, as assessed by oxygenation (partial pressure of oxygen/inspiratory fraction of oxygen [PaO(2)/FiO(2)] 326 +/- 63 mm Hg vs 218 +/- 84; P=.083 low vs high, respectively), and lung edema (wet/dry ratio 6.5 +/- 0.7 vs 8.6 +/- 0.9; P=.012). Glucose consumption correlated with wet to dry ratio (R(2)=0.663; P=.014). CONCLUSIONS: We found that the worse the lung function, the greater the consumption of glucose during EVLP. This observation suggests the need to explore lung metabolism during EVLP to possibly obtain metrics for evaluation. PMID- 21620035 TI - A new liver autotransplantation technique using subnormothermic machine perfusion for organ preservation in a porcine model. AB - BACKGROUND: Hepatic resection is the gold standard of therapy for primary and secondary liver tumors, but few patients are eligible for this procedure because of the extent of their neoplasms. Improvements in surgical experience of liver transplantation (OLT), hepatic resection and preservation with sub-normothermic machine perfusion (MP) have prompted the development of a new model of large animal autotransplantation. METHODS: Landrace pigs were used in this experiment. After intubation, hepatectomy was performed according to the classic technique. The intrahepatic caval vein was replaced with a homologous tract of porcine thoracic aorta. The liver was perfused with hypothermic Celsior solution followed by MP at 20 degrees C with oxygenated Krebs solution. An hepatectomy was performed during the period of preservation, which lasted 120 minutes, then the liver was reimplanted into the same animal in a 90 degrees counterclockwise rotated position. The anastomoses were performed in the classic sequence. Samples of intravascular fluid, blood and liver biopsies were obtained at the end of the period of preservation in MP and again at 1 and 3 hours after liver reperfusion to evaluate graft function and microscopic damage. RESULTS: All animals survived the procedure. The peak of aspartate aminotransferase was recorded 60 minutes after reperfusion and the peak of alanine aminotransferase and lactate dehydrogenase after 180 minutes. Histopathologic examination under the light microscope identified no necrosis or congestion. Intraoperative echo-color Doppler documented good patency of the anastomosis and normal venous drainage. CONCLUSION: This system made it possible to perform hepatic resections and vascular reconstructions ex situ while preserving the organ with mechanical perfusion (ex vivo, ex situ surgery). Improving surgical techniques regarding autotransplantation and our understanding of ischemia-reperfusion damage may enable the development of interesting scenarios for aggressive surgical treatment or radiochemotherapy options to treat primary and secondary liver tumors unsuitable for conventional in situ surgery. PMID- 21620036 TI - Incidence of cancer in kidney transplantation waiting list patients: a single center experience. AB - INTRODUCTION: It is widely accepted that the risk of malignancies is significantly increased among patients with end-stage kidney disease (ESKD) and after kidney transplantation compared with the general population. Only a few data are available on kidney transplantation waiting list patients. The aim of this study was to investigate solid organ cancer incidence among subjects on the waiting list at a single center. MATERIALS AND METHODS: We retrospectively reviewed the records of all patients enrolled on our kidney transplantation waiting list between August 1, 2008 and July 31, 2010, seeking to evaluate the causes of withdrawal from the list, incidence of cancer, type of neoplasm, and its correlation with clinical features. We estimated the ratio of observed to expected numbers of cancers, the standardized incidence ratio (SIR). RESULTS: Among 1184 patients, we excluded 569 patients from the waiting list including 26 (4.56%) who displayed malignancies. The overall incidence of cancer was 0.11 events/person-months and the overall prevalence of cancer was 2.2%. In 97% of patients, the malignant disease was confined to the primitive organ of origin without secondary dissemination. We observed a prevalence of cancers related to ESKD (17; 65.38%). The SIR for all cancer types in our population compared with the general population was 2.22. The SIR for native kidney and thyroid cancers among our population compared with the general population was >10. CONCLUSION: The incidence of cancer was significantly increased among kidney transplantation waiting list patients compared with the general population. Our study highlighted the importance of a careful, targeted neoplastic screening. It could be particularly important for ESKD-related malignancies like native kidney tumors or thyroid cancers. PMID- 21620037 TI - Preoperative psychological factors predicting graft rejection in patients undergoing kidney transplant: a pilot study. AB - BACKGROUND: The aim of this study was to investigate whether pretransplant psychological variables included in the CBA 2.0 Primary Scale-fear, personality, obsessive-compulsive symptoms, state and trait anxiety, psychological reactions, and depression-could predict graft rejection among patients undergoing kidney transplantation. METHODS: After ethical committee approval we enrolled 33 consecutive adult patients undergoing kidney transplantation. The inclusion criteria were a stable clinical situation in an out-of-hospital setting; Italian language literacy; a minimum of secondary school-level education, and written informed consent. We excluded patients with a psychotic disturbance, neurocognitive deficit, dementia, serious mental delay (IQ <50), current alcohol or drug abuse, recent ideation or attempted suicide or nonadherence to the therapeutic protocol. Acute and/or chronic graft rejection was diagnosed according to clinical and histopathologic criteria. CBA-2,0 "Primary Scale" series of questionnaires were handed out to patients at the time of the examinations to discrem eligibility for transplantation. Analyses of variance were performed to compare psychological scores among patients with versus without graft rejection. Logistic regression analyses of psychological variables were performed to detect possible predictors for graft rejection. The results of the analysis showed that higher psychoticism scores were able to predict graft rejection (P<.05). RESULTS: The findings of this study suggest that it is mandatory to preoperatively plan an holistic treatment including psychological intervention mainly focused on psychoticism. PMID- 21620038 TI - Once-a-day administration of everolimus is safe in de novo renal transplant recipients: 1-year results of a pilot study. AB - INTRODUCTION: The half-life of everolimus is approximately 28 hours, but everolimus is generally administered twice a day. The aim of this prospective, single-center, exploratory study was to compare the efficacy and safety of a once a day everolimus (OD) with the standard twice a day administration regimen (BID) as immunosuppressive therapy in renal transplantation. METHODS: Forty-one de novo renal transplant recipients prospectively assigned to OD (n=21) or BID (n=20) treatment were followed for 1 year. In the OD group, everolimus was orally administered targeting a trough blood level of 2 to 5 ng/mL. In the BID group, everolimus was given twice a day targeting a trough blood level of 3 to 12 ng/mL. All patients also received induction with basiliximab and low-dose calcineurin inhibitor immunosuppression. RESULTS: At 1 year follow-up patient and graft survivals were 100%. The intention-to-treat analysis showed similar renal function between the two regimens: serum creatinine values for OD 1.54 +/- 0.6 versus BID 1.48 +/- 0.53 mg/dL (P=NS). Also the occurrence of acute rejection episodes was not significantly different: 4.8% in the OD versus 15% in the BID group, (P=NS). The median trough blood levels were significantly lower among the OD group: OD 4.5 versus BID 7.2 ng/mL (P<.001). DISCUSSION: This study demonstrated that once a day administration of everolimus achieved excellent patient and graft survival and good renal function without an increased incidence of acute rejection episodes. PMID- 21620039 TI - "Immuknow" to measurement of cell-mediated immunity in renal transplant recipients undergoing short-term evaluation. AB - The aim of this preliminary, observational study was to evaluate the value of ImmuKnow (IK), a new tool to measure the net state of immunefunction among renal transplant recipients, in correlation with clinical and laboratory data among unselected renal transplant recipients. Forty-nine recipients of mean age of 51 years were enrolled and followed for 1 year after transplantation. All subjects received the same immunosuppressive strategy with basiliximab induction and tacrolimus, mycophenolate mofetil and steroid maintenance therapy. Samples for IK were collected before transplantation as well as at 7, 14, 21 and 42 days and after 3, 6, and 12 months. There were 54 samples with IK <225 ng/mL, 201 samples with normal IK values, and 135 samples with >525 ng/mL. We divided recipients into 3 groups with respect to their basal IK values: Group 1 (Gr1; IK <225 ng/mL); Group 2 (Gr2; normal values of IK between 226 and 524 ng/mL); and Group 3 (Gr3; IK >525 ng/mL). At 1 year, we observed a significant difference among IK values at the start and the end of the study: Gr1 vs Gr2, P<.0001; Gr2 vs Gr3, P<.06 and Gr 1 vs Gr 3, P<.01). We observed reduced IK values to predict an increased risk of infection, particularly with cytomegalovirus (CMV) replication while higher IK value did not correlate with an increased risk of acute rejection episodes. Reduction of serum creatine levels occurred within 1 year in all groups (P<.005), but there was a significant difference between Gr 2 versus Grs 1 and 3 (P<.0001 and P<.0005, respectively). There findings suggested that more stable IK values were associated with clinical quiescence and laboratory stability. In conclusion, our preliminary analysis showed a beneficial capacity of this assay to represent the global depression of the immune system. We noted that reduced IK values, as a sign of excessive immunosuppressive therapy, were associated with an increased risk of infection. We did not confirm the predictive value of higher IK values for an increased risk of an acute rejection episode. PMID- 21620040 TI - Interleukin-2 profiles shortly after tacrolimus conversion from a twice-daily to once-daily regimen. AB - A number of studies have indicated that kidney recipients can be safely converted from the twice-daily formulation (Tac-T) to the same dose of a once-daily tacrolimus (TAC) regimen (Tac-O) based upon monitoring of renal function. Conversion from Tac-T to Tac-O is commonly followed by a reduction in Tac trough levels, estimated by some authors to be about 20%. These alterations seem to not be associated with a modification of graft function, but study of inflammatory cytokines would be useful. The aims of our study were to monitor Tac, C-reactive protein (CRP), and interleukin (IL)-2 levels as well as to evaluate renal function among stable renal transplant patients converted from a Tac-T to a Tac-O regimen. We enrolled 10 consecutive stable kidney transplanted patients. Tac trough levels, serum creatinine concentrations, glomerular filtration rates using the Modification of Diet in Renal Disease formula (MDRD), CRP, and clinical assessment were performed monthly for 6 months before and 3 months after the conversion. After conversion we observed a slight but not significant reduction in Tac trough level. Renal function evaluated by serum creatinine and MDRD as well as CRP were not significantly different after conversion. IL-2 levels remained stable after conversion. We identified a group of patients showing reduced Tac trough levels below the therapeutic range and a group with stable Tac levels. No significant differences were observed among the two groups before versus after the conversion. Our results did not show a modification of IL-2, CRP and renal function levels, at 3 months after conversion despite the lower Tac trough concentrations. The clinical meaning of Tac trough alterations is not clear. They might reflect inter- and intraindividual differences in the clearance of Tac as recently described. They did not seem to be associated with activation of an inflammatory pathway. PMID- 21620041 TI - Effects of switching from twice-daily to once-daily tacrolimus formulation on quality of life, anxiety, and transplant benefit perception after kidney transplantation. AB - OBJECTIVE: This study investigated whether switching from the twice-daily (Prograf; TAC) to the once-daily formulation of tacrolimus with extended release (Advagraf; XL) affected quality of life, anxiety, and transplant benefit perception after allogeneic kidney transplantation. METHODS: After local Institutional Review Board approval, 78 adult patients prescribed twice-daily tacrolimus for >=1 year after kidney transplantation were asked to participate in this study. All patients were evaluated at T0 (before the switch), and the 49 who accepted the change were reassessed after 6 months (T1). The following tests were used: (State and Trait Anxiety Inventories Y1 and Y2, (Psychologic General Well Being Index), and modified Transplant Effect Questionnaire for posttransplantation symptoms. Blood samples for laboratory profiles and determinations of drug concentrations were obtained throughout the study period. RESULTS: There were no significant differences between the psychologic variables at T0 among patients who switched from TAC to XL (n=49) versus those who did not participate (n=29). Eight of the 49 patients who accepted the drug conversion were reswitched to TAC because of adverse events. At T1, the remaining switched patients (n=41) showed an increase in the disclosure of having undergone transplantation (P<.05) versus nonswitched patients; whereas reswitched patients (n=8) showed less positivity and well-being (P<.05) compared with those who remained in the switched regimen. CONCLUSION: The findings suggested increased disclosure of having undergone transplantation among patients who decided to switch from TAC to XL. PMID- 21620042 TI - Tacrolimus trough levels and level-to-dose ratio in stable renal transplant patients converted to a once-daily regimen. AB - Numerous evidence has been reported to support a safe 1:1 conversion from the twice-daily tacrolimus (Tac-T) to the once-daily tacrolimus regimen (Tac-O), but frequently there is a reduction in drug trough levels, which has been estimated by some authors to be about 20%. The relationship between Tac-O dosage and trough levels after conversion is not clear. The tacrolimus trough levels-to-dose ratio has been applied to better define the wide variability in doses and blood levels of tacrolimus. The aim of this study was to evaluate tacrolimus trough levels, tacrolimus daily dosage, and tacrolimus level-to-dose ratio during 1 year pre postconversion follow-up in 31 stable kidney transplant patients who had received Tac-T therapy for over 6 months with stable renal function. They were converted to the same dosage of Tac-O. Patients before and after conversion were their own controls. The trough levels of tacrolimus showed a slight albeit significant reduction after conversion, remaining in the therapeutic range. Nineteen percent underwent an adjustment in total daily dosage after conversion versus 39% before conversion with no significant difference. No significant differences were detected in the total daily dose administered either by tacrolimus level-to-dose ratio before or after conversion. Kidney transplant recipients under Tac-O therapy were safely maintained using the same therapeutic monitoring as when receiving Tac-T. PMID- 21620043 TI - Switch from twice-daily tacrolimus (Prograf) to once-daily prolonged-release tacrolimus (Advagraf) in kidney transplantation. AB - Advagraf is a new modified-release once-daily formulation of tacrolimus. The aim of this study was to define the efficacy and safety of switching from Prograf to Advagraf immunosuppression in kidney transplant recipients. The switched dose ratio of Prograf to Advagraf was 1:1. Forty-one patients (34 men and 7 women) were switched at 36.6 +/- 16.1 months after kidney transplantation. All patients maintained stable renal function and the conversion. In 16 subjects it was possible to withdraw steroid administration after obtaining adequate Advagraf blood levels, among whom 14 remained steroid free. Adverse events, including dizziness and tinnitus, were reported in 1 patient, who was reverted to Prograf. One patient who was receiving triple therapy with low tacrolimus blood levels experienced are acute rejection episode. The switch to Advagraf was safe and efficacious in kidney transplant recipients with or without steroid administration. Moreover, interruption of steroid was possible and well tolerated after achieving adequate stable blood levels with Advagraf. PMID- 21620044 TI - Survival after kidney transplantation does not differ with 50-59- or over 60-year old expanded-criteria donors. AB - BACKGROUND: Use of expanded-criteria donors (ECDs) for kidney transplantation has progressively increased in the past years with the intent to improve the number of available grafts. However, it is still uncertain if ECD kidneys have worse survivals than standard-criteria ones. The aim of this study was to retrospectively analyze a cohort of ECD patients comparing the 2 subgroups of 50 59- and >60-year-old donors in terms of donor, recipient, and transplant features and survival rates. METHODS: Ninety-one cases were analyzed. The cohort was stratified into 2 subgroups according to donor age: group 1, age 50-59 years (n=26); and group 2, age >=60 years (n=67). RESULTS: Group 2 represented older donors and a higher percentage of donors with a previous history of hypertension. In Group 1, preharvest creatinine values showed higher results. No difference was detected regarding patient and graft survivals, with 5-year survival rates of 92.3% versus 86.7%, and 70.8% versus 69.8%, respectively. The best way to select the donors is still under debate. In our experience, a biopsy-driven selection was performed exclusively for group 2 ECDs. Considering the similar survivals obtained, we speculated that an accurate biopsy-based selection of older grafts allows one to avoid "bad" donors from the allocation system, thereby obtaining improved survival results. CONCLUSIONS: Biopsy-driven pretransplantation selection appears to be the main system to optimize results, to achieve outcomes similar to nonbiopsied younger donors. Routine biopsies also in the younger subgroup of ECD may achieve a further improvement in survival. PMID- 21620045 TI - Kidney transplantation from older donors. AB - BACKGROUND: The use of kidneys from older donors has become generally accepted and increasingly common, despite the knowledge that donor age is a well-known risk factor for graft failure. AIM: To review our experience with the utilization of kidneys from donors older than 60 years. PATIENTS AND METHODS: Among two hundred eight patients, 32 (group A) received an organ obtained from a donor older than 60 years. The organs were age-matched with a maximum gap of 20 years between donors and recipients. Organs from older donors were assigned to recipients presenting a body mass index lower than that of the donor. The primary end point was patient and graft survival. Secondary endpoints were incidences of delayed graft function and of acute rejection episodes as well as renal function at 3 months and yearly. RESULTS: The two groups were comparable in terms of demographic features, indications for transplantation, comorbidities, as well as cold and warm ischemia times. The Mean lengths of follow up were 31.4 +/- 20.3 months and 30.3 +/- 20.1 months, respectively. Graft and patient survivals were comparable. Mean creatinine values at the study intervals were significantly lower among group B who received grafts from younger donors. The incidence of delayed graft function and acute rejection episodes were similar: 15.6% (5/32) versus 20.5% (36/176; P=0.35) and 15.6% (5/32) and 12.1% (21/167; P=0.136) in groups A and B, respectively. CONCLUSIONS: Donor age older than 60 years showed a negative impact on kidney function. Though, given the escalating disparity between organ supply and demand, this precious source of organs cannot be neglected. We need better ways to use the available organs. PMID- 21620046 TI - Increasing levels of hemoglobin improve renal transplantation outcomes. AB - INTRODUCTION: Several studies have reported various data on prevalence of posttransplant anemia (PTA). We have little information about its impact on long term graft outcomes and few studies of the optimal hemoglobin (Hb) target in kidney transplantation. METHODS: We examined retrospectively 144 kidney transplant recipients of mean age 44.4 +/- 12.3 years and follow-up of 40.5 +/- 4.6 months. Exclusion criteria were age below 18 years, multiorgan transplantation, and graft failure in the first year. Using simple and multiple linear regression models, we evaluated the potential prediction of a serum concentration of Hb at 1 year after renal transplantation on allograft outcome as measured by Delta% estimated glomerular filtration rate (eGFR), the difference between eGFR, measured with the Modification of Diet in Renal Disease (MDRD) formula, at the end of follow-up, and at 1 year. Multiple models were adjusted for recipient sex, recipient age, donor age, ESA therapy, acute rejection episodes (ARE), days of delayed graft function, human leukocyte antigen mismatches and cold ischemia time. RESULTS: At 1 year after transplantation, the mean Hb level was 13.77 +/- 1.87 g/dL in males and 12.52 +/- 1.53 g/dL in females. The average eGFR at 1 year was 63.07 +/- 25.88 mL/min. At the end of follow-up, the mean Delta% eGFR was -5.73% +/- 27.30%. Blood concentration of Hb correlated with donor, recipient sex, ARE, and eGFR at 1 year. There was a close correlation between the Delta% Hb and eGFR upon univariate analysis and the multiple linear regression model. Hb was the only predictor of transplant outcome. CONCLUSIONS: Many factors are involved in kidney allograft function. Among these, Hb is important. In this work we demonstrated that increasing levels of Hb at 1 year after transplantation seemed to predict better preservation of graft function, representing a marker of a good quality graft. PMID- 21620047 TI - Multimodal therapy with combined plasmapheresis, photoapheresis, and intravenous immunoglobulin for acute antibody-mediated renal transplant rejection: a 2-year follow-up. AB - BACKGROUND: Recently, the role of antibodies has been documented in the development of acute rejection episodes. Antibody-mediated acute rejection (AMAR) may develop at any time after transplantation, with an incidence of almost 7%. Several therapeutic approaches have been proposed in the past decades. However, no data exist regarding combined plasma treatment (PT) and extracorporeal photopheresis (ECP). The aim of this study was to report an initial single-center experience of combined PT and ECP with high-dose intravenous immunoglobulin (IVIg) for the treatment of AMAR. METHODS: Three patients were treated with this approach. RESULTS: In 2 cases, we observed immediate restoration of graft function, and in 1 case, in which we interrupted the protocol owing to lack of patient consent, the graft was lost. No organ infections were reported during the therapy period. The rationale for use of ECP is related to the presence of mixed antibody and cell-mediated mechanisms in acute rejection episodes. ECP inhibits specific pathogenic T cells. CONCLUSION: Our approach seemed to give good results in terms of graft survival and safety. PMID- 21620048 TI - Thyroid status and kidney transplantation outcomes. AB - INTRODUCTION: It is known that end-stage renal disease patients can display abnormal thyroid gland function, which may cause autoimmune hypothyroidism or subclinical alterations. The impact of thyroid function on graft outcomes is not completely clear among renal transplant patients. The aim of this study was to evaluate thyroid function among a cohort of 136 consecutive renal recipients in correlation with clinical parameters of graft function. MATERIALS AND METHODS: We performed a cross-sectional study on 136 subjects including 84 males and 52 females of overall mean age of 49.71 +/- 10.98 years who underwent renal transplantations between 2005 and 2009 and had a mean follow-up of 28.3 +/- 15.7 months. All patients were treated with a calcineurin inhibitor, steroids, and mycophenolate mofetil. The exclusion criteria were age below 18 years, multiorgan transplantation, graft failure in the first 6 months, or presence of a thyroid neoplasm. We evaluated levels of serum FT3, FT4, and thyroid-stimulating hormone (TSH) in relation to the following parameters: body mass index (BMI), serum creatinine, estimated glomerular filtration rate estimated glomerular filtration rate (eGFR) by Modification of Diet in Renal Disease (MDRD) formula, proteinuria/24 hours, serum sodium, potassium, calcium, phosphorus, cholesterol, high-density lipoprotein, low-density lipoprotein, triglycerides, and hemoglobin (Hb). RESULTS: Only 6.4% of our transplant recipients were treated with levothyroxine sodium. The patients showed an average FT3 of 3.24 +/- 0.5 mg/dL; average FT4 of 0.84 +/- 0.1 mg/dL, and mean TSH of 1.29 +/- 0.8 mg/dL. The study showed no relationship between thyroid hormones and age of the transplant, while there was a significant difference in FT3 levels between men and women. We also observed a significant correlation between FT3 and serum creatinine, eGFR, serum sodium, BMI, and Hb; whereas there was no correlation with other variables. The correlations between FT4 and TSH and all examined variables were not significant. CONCLUSIONS: The interactions between the thyroid and the kidney have been incompletely studied among patients with renal transplants. Our data showed that the presence of low serum FT3 levels correlated with worse graft function, anemia, BMI, and serum sodium. Thus low FT3 levels could be predictive of graft function, especially in the 5 years posttransplantation. PMID- 21620049 TI - Quality of life in kidney transplantation from marginal donors. AB - OBJECTIVE: Enhancement of the subjective components, recognition of overall needs, and careful consideration of subjectively perceived quality of life among dialysis and/or transplanted patients appear to be key objectives to promote optimized adherence to treatment and active cooperation of the patient. This study explores the relationship between self-rated health among recipients of kidney transplantations from deceased marginal donors (age older than 55 years) in relation to gender, age, time on dialysis, years after transplantation, and donor age. PATIENTS AND METHODS: Posttransplant quality of life was assessed with the Complete Form Health Survey (SF-36) in 70 recipients of kidney transplantations from marginal deceased donors. RESULTS: Donor age did not negatively influence health status perceived by the subjects. The vitality and mental health seemed to increase with greater donor ages, but the status of perceived health, vitality, social activities, and mental health were negatively influenced by the age of the transplant. CONCLUSIONS: Our study shows that good general health and social well-functioning can be achieved also among recipients of organs from older donors. Thus, age alone should not be a barrier to organ donation, providing that the organ function is normal and that specific disease is absent in the organ. PMID- 21620050 TI - Polyomavirus BK replication in adult polycystic kidney disease post-renal transplant patients and possible role of cellular permissivity. AB - Cell division and differentiation but not proliferation seem to be necessary for BK virus (BKV) replication and reactivation of persistent infection. Only terminal differentiating cells are permissive to BKV replication. Renal tubular epithelial cells in human adult polycystic kidney disease (ADPKD) are characterized by increased proliferative activity leading to cyst growth with less cellular differentiation. The aim of this study was to evaluate the possibility of different BKV replication patterns in patients with polycystic kidney disease versus non-ADPKD patients. From May 2006 to April 2008, we performed renal transplantations in 47. Eleven patients were affected by ADPKD (Pc group) and 36 patients, non ADPKD (n-Pc group). BKV replication was evaluated by quantitative real-time polymerase chain reactions (PCR) on plasma and urine samples at 12 hours (T0/early) as well as 3 (T3) and 6 (T6) months after transplantation. BKV viremia occurred in 9%, 12.5%, and 20% among the Pc group versus 33.3%, 42.4%, and 50% among the n-Pc group at 12 hours as well as 3 and 6 months posttransplantation, respectively. A higher discordance (BKV-PCR blood /urine +) was observed in plasma and urine BKV replication among Pc versus n-Pc groups. We hypothesized that the lower number of patients with active BKV plasma replication in the Pc group may be related to a lower cellular permissivity of the renal tubular epithelial cells in ADPKD. PMID- 21620051 TI - Combined measurement of serum DNA and urine VP1 messenger RNA in monitoring BK virus replication in kidney graft recipients. AB - Evaluation of BK virus replication is a fundamental tool in the monitoring of renal transplant recipients. Herein, we investigated the role of urine VP1 messenger RNA (mRNA) quantification and combined measurement of serum DNA and urine VP1 mRNA in 428 kidney allograft recipients. BK viremia and viruria were detected in 24 (5.6%) and 54 (12.6%) patients, respectively. A diagnosis of BKV associated nephropathy (BKVAN) was established in 2 patients, both within the first year posttransplantation. Based on urine VP1 mRNA measurement, BKV replication was observed in 10 (2.1%) patients, 2 of whom displayed BKVAN. Urine VP1 mRNA was detected in all cases in association with viremia except 5 and in all cases with viruria. No difference among VP1 mRNA levels was noted between the 2 BKVAN patients and the highest values in patients without BKVAN. The urine VP1 mRNA result by analysis using the operating characteristics was not superior to viremia, despite the improvement obtained with the combined measurement of viremia (cut-off, 16,000 copies/mL) and urine VP1 mRNA (>10,000 copies/10(3) cells). In conclusion, VP1 mRNA measurements may complement viremia and viruria to monitor BKV replication, although its use is limited by its technical complexity in comparison with DNA detection. PMID- 21620052 TI - Evaluation of metabolic acidosis in patients with a kidney graft: comparison of the bicarbonate-based and strong ion-based methods. AB - BACKGROUND: According to the traditional bicarbonate-based approach, metabolic acidosis is highly prevalent in kidney transplant recipients. However, the bicarbonate-based approach has been questioned by intensivists using strong ion difference-based methods. METHODS: We compared the results obtained by the strong ion-based with the traditional approach based on bicarbonate among a cohort of 83 kidney transplant recipients. RESULTS: Fifty-five percent of the patients were acidotic based on venous bicarbonate (<23 mmol/L) and 49% by the use of the effective strong ion difference (SID(effective)) (<37 mmol/L). Bicarbonate and SID(effective) were linearly correlated (r=0.94; P<.0001), with a slope close to 1. A greater percentage of patients presented with an increase in unexplained anions by the strong ion gap (SIG) than by the anion gap corrected (AG(corrected)) method (42 vs 32%, respectively). AG(corrected) and SIG were directly related (r=0.919; P<.0001), but the best fit of the relationship was polynomial with a progressively greater effect on SIG with increased AG(corrected), suggesting that as anions progressively accumulate, their detection by SIG increases. By multiple regression analysis, plasma chloride, potassium, uric acid, and phosphate predicted blood bicarbonate. Analogously, chloride, potassium and uric acid predicted SID(effective). Age was a predictor of changes in AG(corrected), whereas age and plasma urea predicted SIG. CONCLUSIONS: The use of the SID yielded results that were similar to the traditional bicarbonate-based approach. Conversely, SIG appeared to be more sensitive than AG for detection of anion accumulation among patients with a kidney graft. PMID- 21620053 TI - Proteinuria is a predictor of posttransplant anemia. AB - INTRODUCTION: Posttransplant anemia (PTA) involves many factors. Although the link between the hemoglobin (Hb) levels and renal function is known, the relationship between proteinuria and PTA hemoglobin has not been widely explored. The aim of this study was to evaluate whether proteinuria was a predictor of anemia and whether erythropoietin-stimulating agent therapy was a protective factor for kidney damage among transplantation patients. METHODS: We retrospectively examined 144 kidney transplant recipients of mean age 44.4 +/- 12.3 years and a mean follow-up period of 40.5 +/- 4.6 months. Exclusion criteria were age under 18 years, multiorgan transplantation, proteinuria at 6 months over 1.5 g/d, and transplant failure within the first year. Using regression models, we evaluated the potential predictive power of proteinuria at 6 months after renal transplantation for anemia as expressed by Hb levels at 1 year. RESULTS: The frequency of patients with PTA was 38.89% at 1 year, 35.21% at 2 years, and 31.43% at 3 years. Variables with significant correlations with anemia upon univariate analysis were: proteinuria, donor age, acute rejection, estimated glomerular filtration rate, s-creatinine, and salbumin. Upon multivariate regression analysis 24-hour proteinuria and s-albumin remained independent predictors of 1-year PTA. Univariate analysis among the entire cohort showed a significant correlation between 1-year Hb and proteinuria/24 hours at 6 months (P=.007), an observation that was confirmed in the adjusted model along with recipient sex. Patients were then divided into two groups regarding treatment with erythropoiesis stimulating agents (ESA). Multivariate analysis showed that proteinuria (P=.005) was a predictor of Hb only among the group of patients who did no receive erythropoietin, whereas this relationship disappeared among the group treated with ESA. CONCLUSIONS: These results showed that proteinuria at 6 months was a predictor of Hb levels at 1 year. Treatment of transplant patients with ESA may be a protective factor for renal endothelial damage expressed as proteinuria. PMID- 21620054 TI - Metabolic syndrome and cardiovascular disease in kidney transplantation. AB - INTRODUCTION: Metabolic syndrome (MS) is an important cardiovascular risk factor. The aim of this study was to evaluate the incidence of MS in an Italian kidney transplant recipient population and its relationship to the incidence of major adverse cardiovascular events (MACE) after renal transplantation. METHODS: The prevalence of MS was evaluated according to the National Cholesterol Education Program Adult Treatment Panel III criteria among adult recipients who underwent a renal transplant between January 1997 and December 2007. In this period, we prospectively recorded the incidence of MACE to be related to the presence of MS. RESULTS: We included 425 kidney transplant recipients in the study including 62% males and an overall median age 46 years (interquartile range=36-54). The prevalence of MS was 41.2% at 6 months after transplantation and 46.6% at 5 years. During the follow-up (median=5.1 years), 32 patients (7.5%) experienced at least one MACE. The detection of MS at 6 months after transplantation was significantly associated with an increased risk of MACE occurrence (MS IRR=2.2 P=.05). CONCLUSIONS: Our findings indicated that MS was largely present in the transplant population confirming that as in the general population, it was a significant risk factor for the occurrence of severe cardiovascular disease. Early identification and treatment of patients with MS may improve long-term patient survival. PMID- 21620055 TI - Postexplant residual cavity hemostasis with a TachoSil patch. AB - Intracapsular nephrectomy as the standard explant technique for a unfunctional graft is associated with intra- and postoperative complications due mainly to hemorrhage, fluid collections, effusions from damaged tissues and the residual cavity. We have reported herein a positive experience with the use of a collagen medical sponge patch into the surgical site for hemostasis. Use of this device produced a reduction in postoperative bleeding and collections with shortened time of drainage, reduced infection risk, and earlier discharge. PMID- 21620056 TI - The use of a polymeric sealant for prevention of posttransplantation lymphocele. AB - The aim of this study was to investigate the effectiveness of a synthetic polyethy lenglycol (PEG) sealant to prevent lymphocele formation after kidney transplantation. The examined group consisted of 719 recipients including 294 female and 425 male who were immunosuppressed with a calcineurin inhibitor, plus basiliximab since 1999, and with mycophenolate mofetil in addition since 2000. We retrospectively analyzed the incidence of lymphoceles among 545 recipients operated between November 1999 and November 2007 (group I), 93 recipients at standard risk for lymphocele transplanted between December 2007 and December 2009 (group II) in whom we performed only routine ligation of the lymphatic vessels during preparation of the graft, and 31 patients also transplanted between December 2007 and December 2009 who were at higher risk for lymphocele (group III) and underwent an off-label application of the PEG sealant. There was no significant difference in patient demographic features among the groups. In total, 21 group I, patients (3.5%) developed symptoms of a lymphocele that required >=1 corrective procedures, whereas only 1 group II patient (1.07%) developed a lymphocele and no group III patient evidenced a symptomatic lymphocele. No adverse events were observed among group III patients after PEG sealant application. although the preliminary results are interesting, a prospective randomized study is required to assess the cost-effectiveness of PEG sealant to prevent lymphocele formation. PMID- 21620057 TI - Urologic complications in kidney transplantation: a single-center experience. AB - INTRODUCTION: Urologic complications are an important cause of morbidity in kidney transplantation. To prevent this occurrence, several studies have reported the benefit of stenting. The aim of this study was to compare the efficacy of two different types of stents to protect the urinary anastomosis in kidney transplantation. METHODS: We performed a retrospective analysis of 139 kidney transplant recipients who underwent ureteralneocystostomy by the Lich-Gregoire technique between January 1995 and July 2010. On the basis of the type of stent we divided transplant patients into two groups: group A (n=90), the internal external Bracci catheter and group B (n=49), the double-J stent. The urologic complications evaluated in both groups were: urinary tract leakage, obstruction, and infections. We also recorded the duration of the postoperative hospitalization. RESULTS: The incidences of urinary fistulae and ureteral strictures between the two groups were similar (around 3%). A higher incidence of urinary infections, however, was registered among group A compared with group B (46% vs 10%; P<.05). The postoperative hospitalization period was consequently longer in group A then group B (35 +/- 3 vs 24 +/- 2 days; P<.05). CONCLUSION: In our series of kidney transplantations, the occurrence of urinary fistulae and ureteral strictures was not influenced by the type of stent. The use of a double J stent, however, appeared to be associated with a significantly decreased incidence of urinary tract infections leading to shorter postoperative hospitalizations. PMID- 21620058 TI - Intensive care unit admission of decompensated cirrhotic patients: prognostic scoring systems. AB - Cirrhotic patients who need critical care support show high morbidity and mortality rates compared with other critically ill patients. Their prognosis is, in fact, influenced by both the severity of the underlying hepatic disease and the worsening of extrahepatic organ function. Clinicians and investigators have been persistently looking for objective scoring systems capable of providing accurate information on disease severity and short-term prognosis. Risk stratification helps differentiate patients who would not benefit from admission to the intensive care unit (ICU) from those who could achieve better outcomes once aggressively treated. The most common scores, ie, multiple organ dysfunction score, sequential organ failure assessment, and acute physiology and chronic health evaluation, developed in general ICUs to evaluate illness severity, have also been validated to predict the prognosis of cirrhotic patients admitted to the ICU. However, their absolute predictive value has been questioned. A weakness of common prediction models consists in not recognizing the continuum of physiological changes in critically ill decompensated cirrhotic patients. In addition, the predictive power to stratify individual risk is relatively low due to the great variability of liver dysfunction stages, the severity of related manifestations, and the number of nonfunctioning organs on admission. Probability models are not capable of predicting whether a patient will live or die with 100% accuracy, nor can they deny or confirm the indications for mechanical ventilation, vasopressor support or renal replacement therapy, or help to decide when to withhold or withdraw support. Because there are no absolute criteria to predict which cirrhotic decompensated patients will improve with normalization of organ function or deteriorate progressively, a scoring system should be regarded as an adjunct rather than a substitute for clinical judgment in the decision process concerning whether a patient should be admitted to the ICU. PMID- 21620059 TI - Management of sepsis during MARS treatment in acute on chronic liver failure. AB - INTRODUCTION: The aim of our study was a 30-day follow-up of the use of early detection of endotoxin by the endotoxin activity assay (EAA) for patients with acute liver failure superimposed on chronic liver disease (AoCLF) and treated with polymyxin-B hemoperfusion-based (PMX-DHP) treatment and albumin dialysis in the molecular adsorbent recirculating system (MARS). MATERIALS AND METHODS: From February 2008 to July 2010, we evaluated 10 AoCLF patients experiencing systemic inflammatory response syndrome (SIRS) in association with suspected infection and an EAA-positive test (>0.60). These patients awaiting liver transplantation (OLT) showed similar Model End-Stage Liver Disease (MELD) scores (range, 19-25) and encephalopathy grade <= 2. Five patients received therapy to remove endotoxins with PMX-DHP with MARS treatment for liver failure (group A); the other 5 patients received MARS treatment only (group B). RESULTS: Two PMX-DHP treatments were performed in 4 group A patients (average EA=0.66 [range, 0.61-0.70]) and 3 treatments for 1 patient (EA=0.92). All 5 subjects underwent an average of 4 MARS treatments (range, 3-5). At the end of therapy, the median EA level was 0.42 (range, 0.37-0.48). As reported in the literature, we achieved a significant improvement in liver and kidney functions using MARS. Measurements of lactate, interleukin (IL)-6, and tumor necrosis factor (TNF)-alpha were significantly improved among patients treated with the extracorporeal therapies. At 30 days of observation, all 5 patients treated with MARS plus PMX-DHP are alive. In group B, a mean of 7.5 MRAS treatments were performed. We observed an improvement in hemodynamic and liver functions with reduced levels of proinflammatory cytokines and lactates in 4 patients. One patient showed no improvement in clinical status with the development of sepsis and subsequent multiorgan failure after 24 days. CONCLUSION: The possibility of an early diagnosis using the EAA in AoCLF patients could prevent the progression of the sepsis cascade. The use of PMX-DHP and MARS in these patients, could lead to resolution of clinical status in a short time. PMID- 21620060 TI - Microwave thermal ablation for hepatocarcinoma: six liver transplantation cases. AB - Surgical resection for malignant hepatic tumors, especially hepatocarcinoma (HCC), has been demonstrated to increase overall survival; however, the majority of patients are not suitable for resection. Radiofrequency ablation (RFA) is the most widely used modality for radical treatment of small HCC (<3 cm). It improves 5-year survival compared with standard chemotherapy and chemical ablation, allowing down-staging of unresectable hepatic masses. Microwave ablation (MWA) has been extensively applied in Asia and was recently introduced in the United States of America and Europe with excellent results, especially with regard to large unresectable HCC. Our single-center experience between May 2009 and October 2010 included application of MWA to 154 patients of median age +/- standard deviation of 63.5 +/- 8.5 years, 6 males, and 1 female, of mean Model for End Stage Liver Disease (MELD) score (10.1 +/- 3.8). The HCC included, hepatitis C virus (HCV)-related (n=70; 45.5%); alcool (ETOH)-related (n=42; 27%), hepatitis B virus (HBV)-related (n=16; 10.5%); and cryptogenic cases (n=26; 17%). The cases were performed for radical treatment down-staging for multifocal pathology or bridging liver transplantation to orthotopic (OLT) in selected patients with single nodules. A computed tomography (CT) scan was performed at 1 month after the surgical procedure to evalue responses to treatment. Among 6 selected patients who underwent OLT; 5 (83.3%) showed disease-free survival at one-year follow-up. The radical treatment achieved no intraoperative evidence of tumor spread or of pathological signs of active HCC among the explanted liver specimens. In conclusion, a MWA seemed to be a safe novel approach to treat HCC and could serve as a "bridge" to OLT and down-staging for patients with HCC. PMID- 21620061 TI - Necrosis percentage of radiologically treated hepatocellular carcinoma at hepatectomy for liver transplantation. AB - Among a cohort of 414 liver transplantations (OLT) performed form 1996 to 2009, we analyzed 86 patients (20.7%) who were affected by hepatocellular carcinoma (HCC) superimposed on cirrhosis, including 82 with a preoperative diagnosis of tumor; 4 cases had the diagnosis established upon histologic examination after hepatectomy. The gender of 75 patients was male (91.5%), and female in 7 cases (8.5%). The median Model for End-Stage Liver Disease score was 10 (range, 6-23). The underlying liver disease was hepatitis C virus (HCV)-related cirrhosis (41.46%), hepatitis B virus (HBV)-related cirrhosis (15.6%), or alcohol-related cirrhosis (29.3%); cryptogenic; HCV+HIV; HBV+HIV; or HCV+HBV+HIV cirrhosis were present in an other few patients. The diagnosis of HCC and the preoperative staging were defined through radiologic evaluations, without biopsy confirmation in any case. All patients underwent pretransplant radiologic treatments to reduce the drop-out risk while a waiting OLT; OLT was performed for HCC patients within the Milan criteria. Upon histologic examination, the median HCC necrosis was 57 +/- 36%; in 22 cases (26.8%), there were no necrotizing effects. Forty patients (48.8%) display a satisfying degree of disease control with 26 patients (31.7%) downstaged effect; 15 patients (18.3%) showed neoplastic progression with advanced neoplastic disease exceeding the Milan criteria at hepatectomy. One patient had nonevaluable necrosis (1.2%). Our experience showed preoperative radiologic treatments to be not curative but serving as a bridge to OLT. PMID- 21620062 TI - Perioperative intra- and extravascular volume in liver transplant recipients. AB - Assessing adequate volemia to avoid fluid overload and pulmonary edema perioperatively in liver transplantation (LT) is a challenge both for the anesthetist and the intensivist. Volumetric preload indices, such as intrathoracic blood volume index (ITBVI), measured by transpulmonary thermodilution, and continuous end-diastolic volume index (EDVI), measured by pulmonary artery thermodilution, were shown to better reflect preload than central venous pressure (CVP) or pulmonary artery occlusion pressure (PAOP). An ITBVI increase soon after the graft reperfusion influenced pulmonary perfusion without an alteration of extravascular lung water index (EVLWI) and without impaired oxygenation. This study was designed to evaluate relationships between CVP, PAOP, ITBVI, EDVI, and stroke volume index (SVI) within 48 hours after LT. We also investigated the relationship between EVLWI and arterial partial pressure of oxygen and inspired oxygen fraction ratio (PaO(2)/FiO(2)). METHODS: We enrolled 125 patients (103 men and 22 women) undergoing LT. All patients were monitored with the PiCCO system (Pulsion Medical System) and with advanced pulmonary artery catheter connected to the Vigilance System. Hemodynamic volumetric data were collected upon intensive care unit admission and every 8 hours up to 48 hours. Univariate and multivariate regression models were fitted to assess associations between SVI and EDVI, ITBVI, and filling pressures after adjusting for the right ventricular ejection fraction (RVEF, categorized as <=30, 31-40, or >40) and the phase of the observation period. We also assessed associations between PaO(2)/FiO(2) and EVLWI. RESULTS: SVI was associated with EDVI, ITBVI, and RVEF. The models showing the best fit to the data were those including EDVI and ITBVI. Neither CVP nor PAOP showed correlation with SVI. EVLWI inversely correlated with PaO(2)/FiO(2). CONCLUSIONS: In the first 48 hours after LT, ITBVI and EDVI were associated with SVI assessment, whereas CVP and PAOP were not related. EVLWI significantly inversely correlated with PaO(2)/FiO(2). PMID- 21620063 TI - Does caval reconstruction technique affect early graft function after liver transplantation? A preliminary analysis. AB - BACKGROUND: In the past decades, the inferior vena cava (IVC) reconstruction technique has undergone several evolutions, such as biopump, piggyback technique (PB), and laterolateral approach (LLPB). Several advantages are reported comparing the PB technique to biopump use. However, comparison between PB and LLPB has not been as well investigated. The aim of this study was to compare the results in terms of immediate graft function and intermediate graft survival among 3 subgroups characterized by distinct caval reconstruction techniques. METHODS: We retrospectively analyzed a cohort of 200 consecutive adult patients who underwent liver transplantation from January 2001 to December 2009. The patients were stratified according to 3 caval reconstructive techniques: biopump (n=135), PB (n=32) and LLPB (n=33). RESULTS: The LLPB group showed the shortest cold and warm ischemia times and the best immediate postoperative graft function. Survival analysis revealed LLPB patients to present the best 1-year graft survival rates: namely, 90.9% versus 75.0% and 74.1% among the PB and biopump groups, respectively (log-rank tests: LLPB vs biopump: P=.03; LLPB vs PB: P=.05). In our experience, LLPB showed the best graft survivals with an evident reduction in both cold and warm ischemia times. However, it is hard to obtain an irrefutable conclusion owing to the retrospective nature of this study, the small sample, and the different periods in which the groups were transplanted. CONCLUSIONS: LLPB technique was a safe procedure that minimized the sequelal of ischemia-reperfusion damage. This technique yielded results superior to venovenous bypass. No definitive conclusions can to be obtained in this study comparing classic PB or LLPB. PMID- 21620064 TI - Sequential versus contemporaneous portal and arterial reperfusion during liver transplantation. AB - Although sequential portal and arterial revascularization (SPAr) is the most common method of graft reperfusion at liver transplantation (OLT), contemporaneous portal and hepatic artery revascularization (CPAr) has been used to reduce arterial ischemia to the bile ducts. The aim of this study was to prospectively compare SPAr (group 1; n=19) versus CPAr (group 2; n=21) among 40 consecutive OLT from heart-beating donors. There were no differences in the demographics characteristics, Model for End-stage Liver Disease scores, indication for OLT and donor parameters between the groups. OLT was performed using the piggyback technique. The biliary anastomosis was performed in all cases by a duct-to-duct technique with a T-tube in 32% versus 29% of cases without a T tube (P=.83). In the CPAr group, the liver was reperfused simultaneously via the portal vein and hepatic artery. CPAr showed a longer warm ischemia (66 +/- 8 vs 37 +/- 7 minutes; P<.001), while SPAr had a longer arterial ischemia 103 +/- 42 vs 66 +/- 8 minutes (P=.0004). Recovery of graft function was similar. There was no primary nonfunction and delayed graft function occurred among 10% versus 9%. Liver function tests were similar between the two groups up to 90 days case of follow-up- One-year graft and patient survivals were, respectively, 89% and 95% versus 94% and 100% (P=.29). At a median follow-up of 13 +/- 6 versus 14 +/- 7 months, biliary complications included anastomotic stenoses in 15% versus 19% (P=.78) and intrahepatic non-anastomotic biliary strictures in 26% versus none (P=.01) for SPAr and CPAr, respectively. CPAr was safe and feasible, reducing the incidence of intrahepatic biliary strictures by decreasing the duration of arterial ischemia to the intrahepatic bile ducts. PMID- 21620065 TI - Risk factors in liver retransplantation: a single-center experience. AB - Liver retransplantation (Re-OLT) is one of the most debated issues in medicine over the past decade. Re-OLT, currently is accepted for patients with irreversible failure of a hepatic graft caused by primary nonfunction (PNF), hyperacute/chronic rejection, or hepatic artery thrombosis (HAT); whereas it is still controversial for patients with recurrent viral disease, in particular hepatitis C virus (HCV) cirrhosis. Patient and graft survival rates are lower than those observed after primary liver transplantation (OLT). The aim of the present study was to analyze the risk factors that adversely affect survival after Re-OLT in a single center. Medical data were collected for 23 patients who underwent Re-OLT from November 2002 to December 2008 including six men and seven women of mean age of 51.3 years. The most frequent indications for Re-OLT were: PNF (69.5%; 16/23), HCV recurrence (8.6%; 2/23), or HAT (8.6%; 2/23). Mean Model for End-Stage Liver Disease (MELD) at Re-OLT was 27.7 (range = 9-40). After a mean follow-up of 37.4 +/- 30 (standard deviation) months, 43% (10/23) of patients had died, including 70% within the first 2 months after Re-OLT. Sepsis represented the commonest cause of death (40%). Re-OLT was performed for PNF among 90% of succumbing patients. As regards dead patients, 4/10 were HCV(+) whose causes of death were sepsis (n=2), alcoholic cirrhosis (n=2), and undetermined (n=1). Comparing patients who died after liver Re-OLT versus alive patients, we did not find any significant difference in terms of mean MELD (28.6 vs 27; P=NS), MELD > 25 (60% vs 61.5%, P=NS), donor age > 60 years (30% vs 15.3%, P=NS), HCV(+) (40% vs 62%, P = NS), or time interval from OLT to Re-OLT (12.2 vs 777.7 days, P=NS). Patient survivals after Re-OLT were 67% at 3 years and 50% at 5 years, which were lower than those of first transplantations, as reported by other European and International Centers. Forty percent of deaths after Re-OLT occurred among HCV(+) recipients, but for reasons unrelated to HCV infection. PMID- 21620066 TI - University of Modena experience in HIV-positive patients undergoing liver transplantation. AB - INTRODUCTION: Highly effective antiretroviral therapy in the last decade has increased the survival rates of HIV-positive patients, yielding a greater number of HIV patients suffering from liver-related disease. Liver transplantation (LT) is the only curative treatment for end-stage liver disease (ESLD) associated or not with hepatocellular carcinoma (HCC). PATIENTS AND METHODS: From June 2003 to September 2010, 23 patients underwent cadaveric donor LT for ESLD at our institution. Inclusion criteria followed the Italian Protocol for LT in HIV positive patients. Immunosuppressive regimens were based on cyclosporine or tacrolimus, eventually switched to Rapamycin. RESULTS: The median CD4 T-cell count was 275/mmc (range=119-924). All patients were affected by ESLD, which was associated with HCC in 14 cases. Ten patients were within the Milan criteria and four patients exceeded them but were within the San Francisco criteria. Conversion from calcineurin inhibitors (CNI) to rapamycin occurred in ten cases. Hepatitis C virus (HCV) recurrence occurred in 13/21 HCV-positive patients. Acute cellular rejection occurred in eight patients with one developing chronic cellular rejection. Overall patient and graft survivals at 80 months were 50% and 45% respectively. DISCUSSION: LT in HIV-positive patients is a feasible procedure, even if in our experience was burdened by a greater incidence of complications including HCV recurrence and infection compared with HIV-negative patients. PMID- 21620067 TI - Long-term outcomes of orthotopic liver transplantation in human immunodeficiency virus-infected patients and comparison with human immunodeficiency virus-negative cases. AB - Human immunodeficiency virus (HIV) positivity is no longer a contraindication for orthotopic liver transplantation (OLT) due to the efficacy of antiretroviral therapy. The aim of this study was to compare OLT among HIV-positive and HIV negative cohorts; the results were also stratified for hepatitis C virus (HCV) coinfection. Between 2004 and 2009, all HIV-infected patients undergoing OLT from heart-beating deceased donors (n=27) were compared with an HIV-negative cohort (n = 27). The pure HCV infection rate was similar between HIV-positive and HIV negative subjects (63% each). HIV-positive recipients were younger (P=.013). The CD4 count for HIV-positive subjects was 376 +/- 156 at transplantation. The mean model for end-stage liver disease (MELD) score at transplantation was 15 +/- 7 in both groups (P=.92). No differences were observed for donor age (P=.72) or time on the waiting list (P=.56). The median follow-up was 26 (range, 1-64) and 27 months (range, 1-48) for HIV and non-HIV recipients, respectively (P=.85). The estimated 1-, 3-, and 5-year patient and graft survival rates were 88%, 83%, and 83% versus 100%, 73%, and 73% (P=.95), and 92%, 87%, and 87% versus 95%, 88%, and 88% (P=.59) for HIV and non-HIV cases, respectively. HIV/HCV-coinfected patients were younger, namely 47 (range, 40-53) versus 52 years (range, 37-68; P=.003), and displayed lower MELD scores at transplantation compared with HCV-mono infected patients 10 (range, 7-19) versus 17 (range, 8-30) (P=.008). For HIV/HCV coinfected and HCV-mono-infected cases the estimated 1-, 3-, and 5-year patients and graft survival rates were respectively 93%, 76%, and 76% versus 100%, 70%, and 60% (P=.99) and 93%, 84%, and 84% versus 100%, 70%, and 60% (P=.64), respectively. No difference was observed in the histological severity of HCV recurrence. In conclusion, under specific, well-determined conditions, OLT can be a safe, efficacious procedure in HIV patients. PMID- 21620068 TI - Psychological risk factors for graft rejection among liver transplant recipients. AB - The purpose of this prospective study was to find psychological risk factors predicting acute, chronic, and psychological rejection in patients undergoing liver transplantation using Cognitive Behavioural Assessment (CBA-2.0). The primary scale included an assessment of fears, personality, obsessive-compulsive symptoms, state and trait anxiety, psychological reactions, and depression. We prospectively recruited 44 patients undergoing orthotopic liver transplantation (OLT). Exclusion criteria were: education level below secondary school, unstable clinical situation in an out-patient setting, fulminant hepatitis, psychotic disorders, neurocognitive deficits, dementia, serious mental retardation, current alcohol or drug abuse, recent ideation of or attempted suicide, and non-adherence to therapy. CBA-2.0 primary scale series of questionnaires were handed out to patients immediately after the medical examination, which had been performed to ascertain eligibility for OLT. Rejection (acute and/or chronic) was diagnosed according to clinical and histopathological criteria. Psychological rejection was diagnosed when patients declared, after transplantation, a refusal of the new organ which caused psychiatric symptoms requiring medical treatment and/or psychotherapy. Analysis of variance and logistic regression of psychological variables was performed to detect possible risk factors for each type of rejection. A greater fear of repulsive animals was able a predictor for an acute rejection episode (odds ratio=1.1; P<.05). No other psychological pretransplant predictor was noted for chronic or psychological rejection. In patients undergoing OLT, preoperative emotions of fear could predict an acute graft rejection episode. These findings imply that pre-OLT screening should include psychological factors in addition to traditional medical criteria with intervention in selected cases. PMID- 21620069 TI - Causes of sirolimus discontinuation in 97 liver transplant recipients. AB - INTRODUCTION: Sirolimus is a potent immunosuppressant with a mechanism of action different from calcineurin inhibitors (CNIs). It has increasing importance for liver transplant (OLT) patients, in particular if when there is decreased renal function. We evaluated the efficacy and the causes for discontinuation of sirolimus-based immunosuppression among OLT recipients. OBJECTIVE: We retrospectively analyzed 97 liver transplanted patients who were prescribed sirolimus as the principal immunosuppressant. Of these, 61 patients discontinued treatment. Herein we have reported the causes, the timing, and the effects of sirolimus discontinuation. RESULTS: The overall patient survival at 3 years follow-up was 89%. Hepatotoxicity and blood disorders were the most frequent, severe reported side effects. Acute cellular rejection episodes appeared in seven patients and was relieved in 1 to 2 weeks after the sirolimus administration. In 10 patients, the cholestasis associated with chronic rejection was sharply reduced after the introduction of sirolimus. No increase in vascular thrombosis and/or poor wound healing were reported. CONCLUSION: Sirolimus given alone or in combination with CNIs appears to be an effective primary immunosuppressant regimen for OLT patients. However, in the late postoperative period (>3 months) the drug is associated with a relatively high rate of side effects. PMID- 21620070 TI - Role of magnetic resonance imaging in the detection of anastomotic biliary strictures after liver transplantation. AB - INTRODUCTION: Biliary complications after orthotopic liver transplantation (OLT) are the principal cause of morbidity and graft dysfunction, ranging in incidence from 5.8% to 30% of cases. Biliary strictures are the most frequent type of late complication. The aim of this study was to evaluate the role of magnetic resonance cholangiography (MRC) to detect biliary anastomotic strictures among patients undergone OLT with abnormal liver function tests. MATERIALS AND METHODS: One hundred twenty-one of 300 patients who underwent OLT were evaluated by MRC for clinically suspected anastomotic biliary strictures. In all patients, we performed various precholangiographic sequences including T1- and T2-weighted and MRC (radial SE 2D and SS-TSE 3D). Magnetic resonance imaging findings were subdivided as absence or presence of an anastomotic stricture. Diagnostic confirmation was obtained by endoscopic retrograde cholangiography (n=32), percutaneous transhepatic cholangiography (n=21) or surgical treatment (n=18). RESULTS: MRC detected 56 anastomotic biliary strictures, 53 of which were confirmed by other imaging modalities. MRC showed two false-negative cases and three false-positive cases. The sensitivity, specificity, positive and negative predictive values, and accuracy of MRC to detect biliary strictures were 96%, 96%, 95%, 97%, and 96%, respectively. CONCLUSION: MRC proved to be a reliable noninvasive technique to visualize the biliary anastomosis and depict biliary strictures after OLT. MRC should be used when a biliary anastomotic stricture is suspected in an OLT patient. PMID- 21620071 TI - Acute renal failure in liver transplant recipients: role of pretransplantation renal function and 1-year follow-up. AB - Chronic renal failure and acute renal failure (CRF and ARF) are common complications after orthotopic liver transplantation (OLT) that adversely affect patient survival. Many factors influence the development of ARF in the OLT setting. In a previous study we reported an association between ARF and the development of CRF at 1 month after OLT. The aims of our study were to evaluate the influence of ARF on short-, middle-, and long-term renal function after OLT and its influence on 1-year survival of patients and grafts. Fourty-four patients who underwent deceased donor OLT between August 2008 and August 2010 were evaluated pretransplantation, in the perioperative period, and at 1, 6, and 12 months posttransplantation. ARF was associated with CRF at 1 month post-OLT, whereas no association was observed at 6 and 12 months post-OLT. The development of CRF at 6 months post-OLT was associated with pre-OLT renal dysfunction and 1 month post-OLT CRF. Four patients died in the ARF group, whereas 3 patients died in the group without ARF. We confirmed ARF to be a predictive event for short term renal dysfunction. The majority of patients recovered renal function after the first month. Although many pre-, peri-, and post-OLT factors may contribute to the development of posttransplantation CRF, pre-OLT CRF seemed to be the most important risk factor. PMID- 21620072 TI - Model for end-stage liver disease score versus simplified acute physiology score criteria in acute renal failure after liver transplantation. AB - Hepatic function and renal failure are closely related among patients with end stage liver disease (ESLD) due to splanchnic hemodynamic mechanisms that characterize advanced decompensated cirrhosis. Acute renal failure (ARF) is a frequent complication that occurs immediately post-orthotopic liver transplantation (OLT). The Model for End-stage Liver Disease (MELD) score describes the survival of patients with ESLD awaiting OLT related to the severity of liver disease. The Simplified Acute Physiology Score (SAPS II) is a mortality prediction model that scores the severity of illness among intensive care unit patients. In a previous study we observed an association between ARF post-OLT and a higher MELD score, but it was not clear whether this association depends on the grade of ESLD or on the critical condition of liver transplant patients. The aim of this study was to evaluate the association of ARF with MELD score and/or SAPS II criteria among liver transplant patients. We analyzed 46 patients with ESLD who underwent deceased donor OLT. All patients were evaluated at baseline and in the first 7 days post-OLT. According to the RIFLE classification, the incidence of the worst grade of ARF post-OLT was 19.2%. These patients showed significantly higher MELD scores, while there was no association with systemic parameters related to the critical patient's condition or with the mortality score as evaluated by SAPS II criteria. We confirmed the association between renal failure and hepatic function among liver transplant patients. A more severe degree of hepatic dysfunction before OLT was associated with a greater incidence of ARF that can adversely affect patient survival. PMID- 21620073 TI - Polyomavirus BK replication in liver transplant candidates with normal renal function. AB - Polyomavirus-associated nephropathy (PVAN) has a predilection for kidney rather than for other solid organ transplants such as the liver. Immunosuppression is widely recognized to be a major risk factor for PVAN development. Since end-stage liver disease (ESLD) patients are immunocompromised and immunosuppression is a major cause of BK virus reactivation, we sought to evaluate BK virus replication in patients listed for liver transplantation. From April to October 2010, we enrolled 20 patients listed for liver transplantation. BK virus load was measured by quantitative real-time polymerase chain reaction on plasma and urine samples. Viremia occurred in only 1 among 20 patients. We hypothesized that in ESLD patients, the low prevalence of BK virus infection may be related to the prevalent impairment of antibacterial immunity rather than to the viral-specific one. In BK virus reactivation, not only the immunodepressive state itself, but also the specific immunologic mechanisms involved may have a role. PMID- 21620074 TI - Acute renal failure and renal replacement therapy in the postoperative period of orthotopic liver transplant patients versus nonelective abdominal surgery patients. AB - Acute renal failure (ARF) often complicates the postoperative period of patients undergoing orthotopic liver transplantation (OLT); it is habitually associated with high mortality rates. Similarly, patients undergoing major nonelective abdominal surgery are prone to ARF because of their frequent preexistent morbidities, abdominal sepsis, and needed for extended surgical procedures. The aim of this study was to evaluate the incidence of ARF and use of renal replacement therapy (RRT) among OLT versus nonelective abdominal surgery patients and associations with clinical outcomes. We studied all the patients admitted to a surgical intensive care unit (ICU) from January 2008 to December 2009 after OLT or nonelective abdominal surgery. The inclusion criteria were an ICU stay of at least 48 hours and without prior end-stage renal failure. OLT patients (n=84) were younger and less severly ill than surgery patients (n=60). ARF occurrence was lower among the OLT (29%) than the surgery group (47%) requiring RRT in 71% and 53% of patients due to ARF, respectively. The ICU mortality of ARF patients in both groups (29% OLT and 51% surgery) were greater than among subjects without ARF (2% and 6%). The occurrence of ARF is common among these two patient groups, and associated with increased risk of death among in surgery (+45%) versus in OLT (+27%) patients. PMID- 21620075 TI - Early tracheostomy following lung transplantation. AB - BACKGROUND: Despite the common use of tracheostomy in lung transplant (LT) patients, little data exist regarding the indications, timing, periprocedural complications, and impact on outcomes of the procedure. METHODS: We retrospectively analyzed some characteristics and timing of all tracheostomies performed in our lung transplant recipients during a 5-year period. RESULTS: Between January 2004 and November 2009, 31 of 126 lung transplant patients (24.6%) underwent a tracheostomy. They included 14 men with a mean age of 42 years (range, 10 to 61 years) and 17 women with a mean age of 45 years (range, 10 to 64 years). Twenty eight patients undergoing a tracheostomy had a prior bilateral sequential LT and 4 had accepted a single lung. Tracheostomy was surgically performed (ST) in 6 of 31 patients (19.3%); percutaneous tracheostomy (PT) techniques were applied for the other 25 (80.6%) cases. The decision to perform a tracheostomy was made within 4 days from LT in 21 of 31 patients (67.7%), within 8 days in 6 (19.3%) and after 10 days for the other 4 (12.9%) cases. There were no major complications during the PT procedures; no conversion to ST, no loss of airway, no paratracheal insertion, and no accidental tracheal extubation. No pneumothorax, pneumomediastinum, hypotension, hypoxemia, or arrythmyas were recorded in the early post-procedural period. The mean post-LT duration of cannulation was 17 days (range, 5 to 72 days). DISCUSSION: An early tracheostomy may be of considerable benefit for the debilitated patient who will likely require prolonged mechanical ventilation because of a complicated intraoperative course and poor recovery of graft function. PT was performed more quickly and was associated with fewer postoperative complications than ST. We recommend an aggressive strategy in the immediate posttransplant period when extubation fails or is delayed for various reasons. PMID- 21620076 TI - Incidence of neoplastic disease following lung transplantation: a 17-year single center experience. AB - OBJECTIVE: Chronic immunosuppressive therapy following solid organ transplantation has been correlated with an increased risk of posttransplantation neoplastic disease (PTND). In this study we evaluated PTND incidence and outcome at our institution over a 17-year period among patients receiving lung transplantation. MATERIALS AND METHODS: Between February 1992 and December 2008, we performed 290 lung transplantations in 280 patients, including 139 single (48% with 5 retransplantations), and 151 double lung transplantations (52% with 5 retransplantations). Among the 280 patients, 2 had undergone previous double lung transplantation in other hospitals. Follow-up of transplant recipients was performed up to December 2009. RESULTS: Forty-two patients died in the hospital, producing a cumulative early (30-day) mortality rate of 15%. Among the 238 patients discharged from the hospital who entered our follow-up program, 36 (15%) experienced PTND. The mean time between transplantation and diagnosis was 47 +/- 42 months, and patients' mean age at time of diagnosis was 55 +/- 14 years. Overall freedom from PTND was 97%, 84%, and 73% at 1, 5, and 10 years, respectively. PTND was considered to be the direct cause of death in 11 patients (30%). Overall survival of patients with PTND at five years (45%) did not differ from the remainder of the transplanted population (46%). However, PTND became a relevant cause of death in the long-term (>5 years) follow-up. CONCLUSION: Our experience confirms that PTND was frequently diagnosed following lung transplantation. Even if PTND did not seem to significantly affect the survival of patients undergoing lung transplantation, it may become a significant cause of death among those surviving beyond 5 years. PMID- 21620077 TI - Evaluation and significance of cytomegalovirus-specific cellular immune response in lung transplant recipients. AB - In lung transplant recipients, cytomegalovirus (CMV) has been associated with direct ie, organ and systemic infection/disease, and indirect effects, including predisposition to develop acute rejection episodes and chronic allograft dysfunction. Cellular immune responses have been demonstrated to play a role in the control of CMV replication. We evaluated CMV-specific cellular responses among lung transplant recipients associated with the onset of organ infection/disease. Cellular responses were evaluated by an Elispot assay of 48 specimens from 24 patients. All samples were evaluated beyond 1 year after transplantation; CMV DNA was concomitantly detected in bronchoalveolar lavage (BAL) and whole blood specimens. Each patient received a combined prolonged antiviral prophylaxis with CMV Ig for 12 months and gancyclovir or valgancyclovir for 3 weeks after postoperative day 21. Nine patients (37.5%) showed transient or persistent CMV nonresponses including donor-recipient negative serologic matching in 2 cases. Positive CMV DNA results were observed in 18/48 BAL specimens (37.5%) from 12 patients (50%). A viral load of >10(4) copies/mL was observed in only 3 cases, 2 of whom were positive also on whole blood. Among these 3 patients, 2 were responders and BAL (as well as whole blood) specimens collected subsequently were negative for CMV DNA; 1 nonresponder patient exhibited a viral load of 426,492 copies/mL BAL (DNAemia, <2,000 copies/mL), developed CMV pneumonia (confirmed by histopathology and immunohistochemistry) and died within 28 days. The prevalence of CMV DNA positivity on BAL did not differ in relation to the immune response; the mean viral load on BAL showed significantly higher results among nonresponders than responders, namely, 1.4 * 10(5) +/- 2.4 * 10(5) copies/ml versus 7.9 * 10(3) +/- 1.4 * 10(4) (P=.02). Evaluation of CMV-specific cellular immune responses by in vitro immunologic monitoring complements virologic monitoring, helping to identify lung transplant recipients at risk of developing organ infection/disease. PMID- 21620078 TI - Lung transplantation for cystic fibrosis after thoracic surgical procedures. AB - During their life, cystic fibrosis (CF) patients may require thoracic surgical procedures for a number of reasons before undergoing lung transplantation. In the past, this has been considered to be a contraindication to lung transplantation. However, a meticulous surgical technique and careful intraoperative management allows one to perform the transplantation safely. Herein we have reported our experience with CF patients undergoing lung transplantation after previous surgical treatment for pneumothorax or bronchiectasis. PMID- 21620079 TI - The department of psychology within a pediatric cardiac transplant unit. AB - In 1979, the department of Psychology started its activity to provide psychological support to patients and their families during the course of treatment and during the follow-up period. Since 1986, a transplantation program was initiated in the cardiology and pediatric cardiac surgery departments, performing 179 cardiac, 3 heart-kidney, 18 heart-lung 14 both lung, 1 heart-lung liver, and 1 single lung transplantation. From 1993, to September 2010 the kidney transplant program has performed. 218 cases. Since 2008, we performed (32 liver and 1 intestinal transplantations). We examined the quality of life and coping with transplantation attitudes because there is recent evidence of high levels of family anxiety and depression related to a child's poor response to the disease and compliance. Our work associated post-traumatic stress disorder (PTSD) with anxiety and with these clinical variables: gender pathology, rejection, number of drugs frequency of hospital visits, number of hospitalizations, as well as age at and time from transplantation. We examined 56 parents (25 fathers and 31 mothers) of cardiac transplant recipients. Our most relevant data demonstrated the presence of PTSD among 52% of mothers and 40% of fathers. Significant correlations were observed within the trait of anxiety. No correlations were noted between PTSD and other variables. Testing anxiety levels of parents is considered to be basic to provide psychological support to parents, encourage personal skills, and avoid PTSD symptoms. PMID- 21620080 TI - C4d analysis in endomyocardial biopsies of heart transplant patients: is there a correlation with hemodynamic data? AB - BACKGROUND: Endomyocardial biopsy (EMB) is the gold standard for immunologic follow-up to detect acute cellular rejection after cardiac transplantation. Conversely, protocols for the diagnosis and treatment of antibody-mediated rejection (AMR) are not well defined. Histologically, AMR is diagnosed by the presence of capillary damage associated with complement activation. The aim of this study was to correlate C4d expression of activated complement in EMB with hemodynamic compromise upon right heart catheterization. METHODS: Heart transplant patients underwent hemodynamic and histologic follow-up with EMB and right heart catheterization between January 2008 and December 2009 for a total of 491 procedures. The cardiac biopsy was evaluated for acute cellular and AMR by means of the presence of the C4d complement fraction. The histologic results were compared with hemodynamic data registered during right heart catheterization. RESULTS: Comparison of the hemodynamic data of subjects with versus without C4d positivity showed no significant difference. Furthermore, there was no significant difference comparing patients with versus without C4d positivity in the absence of significant acute cellular rejection episodes. (C4d-/ACR- vs C4d+/ACR-). The variation of each single hemodynamic parameter from its basal value (defined as the mean value in case of C4d-/ACR-) seemed to not be influenced by the presence of C4d+. CONCLUSIONS: In our experience, C4d has been routinely evaluated in the majority of EMBs. We could not demonstrate a significant correlation of C4d positivity with hemodynamic compromise. These findings suggest that significant allograft dysfunction is not related to C4d positivity. Therefore, the diagnosis of AMR is difficult to establish, because allograft dysfunction is 1 of the 3 fundamental criteria. PMID- 21620081 TI - A good breath of oxygen for beta-like cells obtained from porcine exocrine pancreatic tissue. AB - Ischemia is the most important factor that affects organ survival during harvesting. The two-layer method (TLM) is one of several cold storage solutions that seeks to preserve organs and cells avoiding in vivo and in vitro ischemia. We compared the retrieval of beta-like elements from exocrine pancreatic cells using TLM versus University of Wisconsin (UW) solutions. For this purpose pancreata laparoscopically harvested from 20 female pigs were preserved in UW solution or TLM before digestion. The resulting exocrine cells were divided into 2 groups: the first was cultured in a designed medium to allow differentiation into beta-like cells and the second was cryopreserved before the differentiation process at -196 degrees C for 8 weeks before culture in the same medium. The results revealed that TLM was better than UW as a preservation solution in terms of beta-cell viability and insulin secretion. We suggest that the use of TLM solution allows one to obtain less damaged cells for research purposes. PMID- 21620082 TI - Glycogen storage disease type Ia and VI associated with hepatocellular carcinoma: two case reports. AB - Glycogen storage diseases (GSD) are inherited metabolic disorders of glycogen metabolism due to intracellular enzyme deficiency resulting in abnormal storage of glycogen in tissues. GSD represents an indication for liver transplantation (OLT) when medical treatment fails to control the metabolic dysfunction and/or there is an high risk of malignant transformation of hepatocellular adenomas (HCA). Herein we have reported two cases of GSD, type Ia and type VI, which were both associated with rapidly growing HCA, and underwent OLT because of suspect changes in their radiological features. Final histological findings in the explanted liver showed the presence of hepatocellular carcinoma (HCC) in both cases. In GSD type Ia and VI, OLT is considered to be the treatment of choice when a liver neoplasm is suspected. While the association of HCC with GSD type Ia is well known, this is the first case of HCC in GSD type VI so far reported to the best of our knowledge. PMID- 21620083 TI - Fulminant multiorgan failure due to varicella zoster virus and HHV6 in an immunocompetent adult patient, and anhepatia. AB - Varicella is a well-known contagious disease of childhood that can also affect both immunodepressed and immunocompetent adults. The present observations concern a previously healthy adult patient who presented with a fulminant hepatitis evolving in multiorgan failure (MOF), associated with an atypical papulo ethemateous cutaneous rash without fever. An hepatic biopsy showed massive necrosis. Because of the persistent MOF and severe hemodynamic instability, total hepatectomy was performed as a bridge to urgent liver transplantation (OLT). Despite temporary improvement, the patients condition progressively deteriorated and he died 11 hours after the hepatectomy, i.e. 7 days after admission to the intensive care unit. High viral loads of varicella zoster virus (VZV) and human herpes virus 6 (HHV6) were demonstrated in the blood and in DNA at post mortem examination of the liver, kidneys, lung, and heart. We hypothesize that VZV infection may occasionally occur in immunocompetent patients due to extremely virulent strains that can be rapidly fatal. The clinical influence of simultaneous infection with HHV6 is not clear. Moreover, the role of a previous steroid treatment as a trigger for a temporary immunodepressed state must be considered. The diagnosis of liver disease from VZV should always be clinically suspected in the presence of concurrent atypical skin lesions and a temporarily immunocompromised state. Therapy with acyclovir was ineffective in our patient. Based on the wide spectrum of VZV infections, fulminant MOF in immunocompetent adults must raise the possibility of VZV with simultaneous HHV6 infection with early listing of the patient for a urgent OLT, possibly with a total hepatectomy as a bridge, due to the therapeutic uncertainty of medical treatments. PMID- 21620084 TI - Neoadjuvant therapy protocol and liver transplantation in combination with pancreatoduodenectomy for the treatment of hilar cholangiocarcinoma occurring in a case of primary sclerosing cholangitis: case report with a more than 8-year disease-free survival. AB - Cholangiocarcinoma has historically represented a major contraindication to liver transplantation at many centers because of its high recurrence rate and low disease-free survival rate, even after radical surgery. Novel neoadjuvant therapy protocols combined with demolitive surgery and liver transplantation seem to achieve successful results in terms of overall and disease-free survivals. Surgery frequently seems to be unsatisfactory only for patients also suffering from chronic cirrhosis or end-stage liver disease. We have reported a case of hilar cholangiocarcinoma occurring in a case of primary sclerosing cholangitis treated with neoadjuvant radiochemotherapy and endoscopic brachytherapy, followed by liver transplantation combined with pancreatoduodenectomy, who has survived free of disease for >8 years. PMID- 21620085 TI - Multiorgan retrieval and horseshoe kidney: case report. AB - Horseshoe kidney, a congenital anatomic condition with fusion of the kidney poles, causes the organ to be placed around the aorta usually below the origin of the mesenteric artery. This congenital disorder affects about 1 in 400 people. Retrieval for multiorgan transplantation dissects and canulates major abdominal vessels, aorta and vena cava, below the renal vessels to infuse refrigerated preservation solution. The presence of a horseshoe kidney could be a hazard for the retrieval team when cannulating when following standard techniques. We have described herein a surgical maneuver exposing the anterior surface of the aorta for canulation. We transected the horseshoe kidney in the midline with the use of a linear cutter stapler GIA 60 mm (Ethicon), after previously ligating both kidney pedicles. This technique was safely performed without the need for cannulation through the iliac vessels. PMID- 21620086 TI - Extracorporeal portal vein arterialization in man after extended hepatectomy to prevent acute liver failure: a case report. AB - Experimental studies have shown that increasing the oxygen supply to the liver through portal vein arterialization (PVA) enhances liver regeneration after partial hepatectomy. Moreover, our previous study demonstrated a beneficial effect of an extracorporeal device to increase the oxygenated blood to the liver and to improve the survival rate of animals subjected to subtotal hepatectomy. Herein we have reported a case of PVA through an extracorporeal device to treat a man after extended hepatectomy leading to acute liver failure (ALF). An obese 69 year-old man (body mass index > 35) affected by multiple metastases from colorectal cancer underwent 80% liver resection; at laparotomy, a steatotic liver was evident due to adjuvant chemotherapy. Moreover, the liver experienced 20 minutes of hepatic ischemia during the resection. At the end of resection he underwent extracorporeal PVA treatment. Blood was withdrawn from the femoral artery and returned into the portal venous system through the umbilical vein. An extracorporeal device was interposed between the outflow and inflow to monitor hemodynamic parameters. Starting from operating room each of six treatments lasted 6 hours per day. Serum and liver samples were collected daily. The extracorporeal device was dismounted at the seventh postoperative day. The postoperative course was assessed at 1 month. The PVA-extracorporeal treatment yielded beneficial effects for subtotal hepatectomy by decreasing serum ammonia, transaminases, and total bilirubin concentration. The international normalized ratio recovered rapidly, remaining significantly lower during the entire postoperative period. The ten-day postoperative period was uneventful. The patient was discharged in good health. He is alive and well at the moment. The arterial blood supply in the portal system through the umbilical vein using an extracorporeal device was easily applicable, efficacious, safe, and cost effective. It may represent a novel approach to treat patients with potential ALF after subtotal liver resection. PMID- 21620087 TI - Preemptive liver-kidney transplantation in von Gierke disease: a case report. AB - Type 1a glycogen storage disease (GSD 1a), or von Gierke disease, is a rare, autosomal-recessive disease caused by a deficiency of glucose-6-phosphatase, which leads to glycogen accumulation in the liver, kidney, and intestinal mucosa. Clinical manifestations include hypoglycemia, growth retardation, hepatomegaly, lactic acidemia, hyperlipidemia, and hyperuricemia. Long-term complications include renal disease, gout, osteoporosis, pulmonary hypertension, short stature, and hepatocellular adenomas, which may undergo malignant transformation. Herein we have described the management and the clinical course of a GSD1a patient who underwent simultaneous preemptive liver- kidney transplantation (SPLKT), which solved the liver and renal disease. We confirmed the rapid normalization of glucose metabolism, and correction of hyperlipemia after liver transplantation. In our opinion uremic patients with GSD 1a with or without adenomas must be considered for SPLKT. To our knowledge this is the fifth case of SPLKT and the first preemptive one to be described in the literature. PMID- 21620088 TI - Late lung retransplantation using extracorporeal membrane oxygenation as a bridge: case report. AB - Lung retransplantation is the only therapeutic option for acute and chronic graft failure, but only a few cases have been described to have been performed with extracorporeal membrane oxygenation (ECMO) support. We describe the successful case of a 46-year-old man treated with right lung transplantation and left lung retransplantation supported by venovenous ECMO. Lung retransplantation is the only therapeutic option to treat severe primary graft dysfunction, major technical problems, and refractory chronic rejection following pulmonary transplantation. Despite a number of comprehensive studies on lung retransplantation, only a few works have addressed the use of extracorporeal membrane oxygenation (ECMO) as a bridge to the surgical reoperation. Herein we have presented a patient treated with pulmonary bilateral retransplantation subsequent to ECMO therapy for progressive deterioration of pulmonary function in single lung transplantation. PMID- 21620089 TI - Double endocrine neoplasia in a renal transplant recipient: case report and review of the literature. AB - INTRODUCTION: The incidence of cancer compared for age groups is 3-4 times higher in transplant recipients than the general population. The increased risk is related to immunosuppressive therapy as well as the use of increasingly older donors and recipients. Although cardiovascular disease with a functioning transplant is the leading cause of death (47%), cancer mortality is significant especially among older patients. However, the most frequent posttransplantation cancers relate to hemolymphopoietic organs and skin, whereas the occurrence of solid tumors elsewhere is rare. Herein we have described a rare case of synchronous double malignancy of endocrine organs (thyroid-adrenal) in a young woman who underwent renal transplantation. CASE REPORT: A 37-year-old woman with end-stage renal disease for 18 years underwent transplantation when she was 30 years old with a 17-year-old standard cadaveric donor receiving immunosuppressive therapy with mycophenolate mofetil, cyclosporine, and steroids. Follow-up demonstrated good indices of renal function with negative tumor pathology at 79 months when, at an annual ultrasound monitoring, we found a lesion in the right lobe of the thyroid and left adrenal neoplasm of dubious interpretation. The cytology for the thyroid was highly suspicious of papillary carcinoma, whereas the histological examination after surgery diagnosed a thyroid multifocal papillary microcarcinoma (mpT1NxMx) and an oxyphil cell adrenocortical carcinoma (pT2, N0). RESULTS: Six months after total thyroidectomy with central lymphadenectomy and left kidney and adrenal gland removal the patient showed no evidence of recurrent lesions and stable graft function. CONCLUSIONS: The rare occurrence of solid tumors after transplantation has no known etiopathogenetic relation. Despite the young age of the patient and the double neoplasm that could have produced an unfavorable outcome for the patient and the graft, careful follow-up for tumor pathologies and multidisciplinary management achieved an early diagnosis of both tumors with a surgical eradication without adjuvant therapy, preserving the life of the patient and the function of the graft. PMID- 21620090 TI - Pulmonary tuberculosis in an HIV- and hepatitis C virus-coinfected kidney pancreas transplant recipient: a case report. AB - Tuberculosis (TB) is a serious infection in immunocompromised patients, such as solid organ transplant recipients and HIV-infected patients. The diagnosis and treatment in this population present several challenges because of the aspecific clinical manifestations, the difficulty in diagnosis, and the choice of the most appropriate therapeutic regimen. Therapeutic challenges arise from drug-related toxicities, interactions between immunosuppressive, antiretroviral, and antituberculous drugs. We present a case of primary TB infection that occurred 3 years after transplantation in a HIV-and hepatitis C virus-coinfected kidney pancreas recipient. The infection was successfully treated with no hepatotoxicity or rejection with a non-rifampin-containing regimen. PMID- 21620091 TI - A rare case of herpes simplex type 1 bronchopneumonia associated with cardiomegaly in renal transplantation. AB - INTRODUCTION: We report a rare case of herpes simplex virus (HSV) type 1B in patient with kidney transplant as a possible cause of patient death. CASE REPORT: A 32-year-old renal transplanted Caucasian man was referred for asthenia, fever, anemia, chest pain, cough, dyspnea, myalgias, peripheral edema, acute renal failure, diffuse cutaneus and mucous vesicles, and acute weight gain. The home therapy consisted of tacrolimus, sodic mycophenolate, and steroids. Laboratory data, bronchoscopy, and bronchial mucosal biopsy revealed HSV1B. We administered antiviral and antibiotic agents and reduced tacrolimus with clinical resolution. But after 10 days from discharge, the patient was admitted for acute cardiomegaly. So using ex adiuvantibus criteria we administered antiviral therapy with complete clinical improvement. CONCLUSION: According to the literature, posttransplant HSV1B infection is a rare but severe complication of kidney transplantation associated with poor graft survival and a high mortality. Only an early, accurate diagnosis with efficient treatment permitted resolution of the problem. Our report stresses the difficulty of HSV2B clinical diagnosis and treatment. PMID- 21620092 TI - Saccular aneurysm of graft renal artery: case report. AB - Repair of anatomic anomalies in a donor kidney's vascular structures requires prompt recognition of the aberrant condition and an appropriate repair technique that does not compromise transplant success. Our report describes a case of a saccular aneurysm of the donor kidney, renal artery, which was identified during harvesting and repaired on the back-table using a prosthetic patch derived from the excised lesion to cover the breach. The long-term results were good. PMID- 21620093 TI - New chances for Hungarian transplantation--preface to the 12th Congress of the Hungarian Transplantation Society. AB - The year 2010 was a milestone in the history of transplantation in Hungary. Hungarian politicians became interested in solving the serious problems facing organ transplantation in our country. The State Secretary announced a program to (1) increase waiting lists, (2) raise donor numbers, (3) establish a lung transplant program, (4) promote education and increase the knowledge base regarding transplantation for the public and the medical profession, and finally, (5) to begin the negotiations for Hungary to join Eurotransplant. Joining Eurotransplant has been a priority of the transplant community. Finally, this year saw the Budapest Transplant Center perform 20% of their transplants from living kidney donors, up from 5% historically. PMID- 21620094 TI - Vladimir P. Demikhov, a pioneer of organ transplantation. AB - Vladimir P. Demikhov was born in a Russian peasant family in 1916. As a biology student at The Moscow University in 1937, he constructed a metal artificial heart and maintained the circulation of a dog for 5.5 hours. From 1946, after his military service, he worked in the Surgical Institute of The Moscow Academy of Sciences performing heterotopic heart transplantations in dogs. In 1947, he performed the first orthotopic lung transplant. Later he performed complex cardiothoracic transplantations as well as renal and hepatic transplantations. He restarted his investigations with the artificial heart and performed coronary bypass operations in dogs. In 1954 he performed a head transplantation, for which he gained worldwide infamy. Stalinist propaganda advertised this fact as the superiority of Soviet science. In fact, it was the upper body of a smaller dog to the neck of a bigger one. The two heads could eat and drink separately. But he could not overcome the problems of rejection, so the longest survival was 1 month among 20 such operations. His influence on the pioneers of transplantation is unquestionable. He was an innovative creative man, and many pioneers of transplantation highly appreciated his work. Demikhov contributed to clinical heart and lung transplantation by demonstrating the possibility of their experimental realization; furthermore, he motivated the pioneers of coronary bypass operations with his work. He died in 1998, but before that was honored with a high state award and the "Pioneer Award" of the International Society for Heart and Lung Transplantation. PMID- 21620095 TI - Childbirth after organ transplantation in Hungary. AB - BACKGROUND: The first successfully delivered newborn after organ transplantation was reported in 1963; since then, >14,000 women have delivered after transplantation. Patients with an end-stage organ disease develop fertility disturbances. One year after a successful solid organ transplantation with stable graft function, fertile women can give birth to a child from a medical point of view. Pregnant transplant patients do experience a high risk of graft function worsening, a rejection episode, and opportunistic infections. Furthermore, the medical therapy may influence teratogenicity. METHODS: Between 1974 and September 1, 2010, 5 Hungarian centers performed 6802 solid organ transplantation and lungs were grafted in Vienna, Austria. The organ distribution was: 5971 kidney, 454 liver, 187 heart, 90 combined pancreas-kidney, 5 combined islet-kidney, and 95 lung transplantation. There were no pregnancies among heart, lung, and pancreas recipients. RESULTS: In all, 3.9% of the renal and 14.3% of the fertile liver transplanted women gave birth to children. To wit, 23 kidney recipients delivered 27 healthy children (17 boys and 10 girls). In 4 cases, 2 children were born, twice as twins. Among liver recipients, 8 women delivered 8 healthy babies. There was no hepatitis C or B virus-positive patient among the mothers. There was no graft insufficiency, rejection or birth defect. Transplanted mothers often display toxemia or preeclampsia during pregnancy requiring cesarean section. The relatively higher ratio of liver recipients was perhaps due to the rarer occurrence of extrahepatic organ damage, like diabetic nephropathy or cardiomyopathy, and the reversible nature of hepatorenal or hepatopulmonary syndrome. CONCLUSION: Delivery of a child by a transplanted mother carries an high risk, requiring interdisciplinary cooperation. The quality of life of solid organ recipients can be significantly raised by childbirth under appropriate circumstances. PMID- 21620096 TI - Help, I need to develop communication skills on donation: the "VIDEO" model. AB - BACKGROUND: Information about brain stem death and donation can be influence the consent rate for donation and its psychosocial effects. The aim of this study was to create a "VIDEO" model that could be used to help physicians to develop communication skills. METHODS: A video recorded 32 simulations of family interviews: 16 under-age and 16 adult donors. They were analyzed during 8 courses conducted in 2008 and 2009. During the VIDEO process, the visual presentation was followed by participants (n=192) discussing interactively the donation situation. After the transcription of the video records, family interviews were explored retrospectively regarding informing relatives about brain stem death and donation, typical communication gaps and common questions from families. The data were examined qualitatively and semiquantitatively. We think that teaching can be optimized by our results. RESULTS: A comprehensible explanation about brain stem death was provided to relatives in 65.63% of cases. The consent of the family was more important for the physicians than the application of the law in 93.75%; 78.13% of physicians emphasized altruism to support donation. Remarkable mistakes of communication included using the teams coma and brain stem death interchangeably (9.38%); applying expressions connected with life in the present tense (21.88%) and mechanically kept alive (21.88%); organ-focused behavior such as "organs to be usable" (34.38%). The frequent questions and statements of "relatives" were "heart beats" (100%), "did he really die?" (65.63%), "fear of loss of integrity of the corpse" (59.38%), and "wake up from the coma" (46.88%). DISCUSSION: Interaction with the family requires great preparation. The communication skills of physicians can be developed through the VIDEO model. The results can be integrated into educational programs that consider the particular features of the given country. PMID- 21620097 TI - The significance of the circumaortic left renal vein and other venous variations in laparoscopic living donor nephrectomies. AB - Among the several vascular variation those concerning the venous system of the kidneys show the most significant variability. They often play an important role when it comes to choosing the kidney to be removed for transplantation. Based on our prior studies, we have surveyed these variations. When performing a laparoscopic living donor nephrectomy owing to the limited field of vision and the restricted possibilities for preparation, preoperative radiologic planning is of utmost importance. We evaluated 55 donors who underwent laparoscopic nephrectomies using the 16-section multidetector-row computed tomography angiography. Among the donors who underwent surgeries we observed circumaortic veins (CAV) in three cases, retroaortic veins in 6 cases, multiple renal veins in 10 cases, and a lumbar vein draining into the left renal vein (RV) in 30 cases. In the 2 cases wherein CAVs were discovered, the team decided to use the other kidney. In 1 case, due to a short right RV, we chose the left kidney. The complex development of the CAV that is sometimes difficult to reconstruct in 3D poses a challenge for both the radiologist and the surgeon. PMID- 21620098 TI - Cytomegalovirus seroprevalence among solid organ donors in Hungary: correlations with age, gender, and blood group. AB - BACKGROUND: Cytomegalovirus (CMV) is endemic throughout the world, affecting most of the population, but the seroprevalence of CMV is known to vary among countries. CMV causes a mild infection in persons with intact immunity; however, CMV infection in organ transplantation is associated with significant morbidity and mortality. The present retrospective study was designed to evaluate the age-, gender-, and blood group-adjusted CMV seroprevalence among solid organ donors, representing fairly the overall Hungarian population (according to Hungarian Central Statistic Institute). This information is important for calculating risk factors for CMV-seronegative recipients. No nationwide estimates of CMV seroprevalence in Hungary (as a representative of Eastern Middle Europe) have been published yet. METHODS: We investigated 2070 organ donors for CMV seroprevalence by measuring the CMV-specific immunoglobulin G. The donors were divided into 3 age groups (2-20, 21-50, and 51-70 years old). A study was also conducted on a fourth group consisting of 200 residents from an old age home. CMV seroprevalence differences were searched according to age-, gender- and blood group distribution. RESULTS: The CMV seroprevalence of organ donors is 85% and of all investigated persons is 86%. The age-specific prevalence increases, starting from 72% in the first group to 99% in the fourth group. Seroprevalence of females was found to be significantly higher than of males (P=.0001). CONCLUSION: We have shown that the overall CMV seroprevalence in the Hungarian population is moderately high at 86%. The opportunity for CMV-seronegative recipients to get a graft from a seronegative donor is statistically only 2%. The seroprevalence of the youngest age group is 72% and so it can be concluded that the Hungarian population acquires the infection mainly in childhood or in the early adulthood. Female gender is a risk factor for CMV infection. This fact must be taken into consideration during the planning of patients' follow-up, prophylaxis, and therapy. PMID- 21620099 TI - Treatment of subclinical injuries detected by protocol biopsy improves the long term kidney allograft function: a single center prospective randomized clinical trial. AB - BACKGROUND: The long-term benefit of early treatment of subclinical disorders detected in kidney allografts by protocol biopsy is controversial. We collected 145 protocol biopsies from 113 recipients for comparison with 51 control patients in a single-center, prospective, randomized trial. METHODS: Ultrasound-guided biopsies were performed in recipients with stable renal function. Samples were taken at 3 (n=66) and/or 12 months (n=79) after transplantation. The biopsies were evaluated according to the Banff scheme, and patients were treated based on the diagnosis. Changes in glomerular filtration rate (GFR) were compared with 51 patients who were randomized as a control group. RESULTS: The findings on 38 samples (29%) were considered to be normal. Based on the pathology findings, such as subclinical acute rejection (n=23), calcineurin inhibitor toxicity (n=28), chronic rejection (n=6), and other specific pathologies (n=23), including polyoma virus nephropathy (n=2), induced treatment among 82 recipients (57%). Significantly better graft function was observed at 3-year follow-up among the biopsy group, compared with controls: GFR = 46.0 +/- 13.8 vs 35 +/- 15 mL/min (P=.002). The 5-year graft survival was significantly higher in the biopsy (81%) than in the control (55.6%) group (P=.0012). CONCLUSION: Early detection and treatment of subclinical pathologies improved graft function and long-term survival. Protocol biopsies were a valuable tool for posttransplantation management. PMID- 21620100 TI - H1N1 vaccination in pediatric renal transplant patients. AB - BACKGROUND: Solid organ transplant recipients undergoing immunosuppressive therapy are considered to be at high risk of serious infectious complications. In 2009, a new influenza pandemic caused serious infections and deaths, especially among children and immunocompromised patients. Herein we have reported the safety and efficacy of a single-shot monovalent whole-virus vaccine against H1N1 infection in the pediatric renal transplant population. METHODS: In November and December 2009, we vaccinated 37 renal transplant children and adolescents and measured their antibody responses. Seroprotection, seroconversion, and seroconversion factors were analyzed at 21 days after vaccination. RESULTS: None of the vaccinated patients experienced vaccine-related side effects. None of the patients had an H1N1 influenza infection after vaccination. All of the patients showed elevations in antibody titer at 21 days after vaccination. In contrast, only 29.72% of the patients achieved a safe seroprotection level and only 18.75% a safe seroconversion rate. More intense immunosuppressive treatment displayed negative effect on seroprotection and seroconversion, and antibody production significantly increased with age. No other factor was observed to influence seroprotection. CONCLUSIONS: We recommend vaccination of children and adolescent renal transplant recipients against H1N1 virus. However, a single shot of vaccine may not be sufficient; to achieve seroprotection, a booster vaccination and measurement of the antibody response are needed to assure protection of our patients. PMID- 21620101 TI - Diagnosis and classification of hemolytic uremic syndrome: the Hungarian experience. AB - BACKGROUND: Hemolytic uremic syndrome (HUS) is a rare disease with various etiologies, making the identification of the specific forms and appropriate treatment difficult. Therefore, clinical and laboratory data from these patients need to be analyzed in national and international registries. Herein we have described 47 Hungarian HUS patients with detailed laboratory and clinical data obtained between 2008 and 2010. METHODS: Blood samples and clinical data of 47 patients with HUS diagnosed according to characteristic clinical signs were submitted for diagnostic evaluation, including complement protein and genetic analysis, measurement of ADAMTS13 activity and antibody analysis against O157LPS and factor H. RESULTS: There were 8 patients with typical diarrhea-positive HUS; 13 with atypical HUS (aHUS) and 26 with secondary HUS/thrombotic thrombocytopenic purpura group characterized by signs of complement consumption and decreased ADAMTS13 activity. Thus, decreased total alternative pathway activity is a promising diagnostic parameter with good sensitivity for aHUS. CONCLUSIONS: These observations highlight the requirement for multiple diagnostic tests together with clinical data to identify the specific cause of HUS. Because the long-term prognosis of aHUS, eg, graft survival after renal transplantation, may vary according to the molecular etiology, it is important for all affected patients to undergo a detailed molecular diagnosis of the disease. There is a clear clinical need for the development and application of novel assay in this field to allow more rapid efficient diagnosis of patients who undergo a first episode of HUS. PMID- 21620102 TI - A single-center experience with kidney transplantation in the verteberal, anal, cardiac, tracheoesophageal, renal, and limb birth detects (VACTERL) association. AB - VACTERL association is a nonrandom association of birth defects in vertebral, anal, cardiac, tracheoesophageal, renal, and limb structures. Renal anomalies are observed in ~60%-90% of VACTERL patients. We present 3 cases to demonstrate the clinical and surgical challenges that these patients present for renal transplantation. One pediatric and 2 adult patients with the VACTERL association were transplanted at a single center; their follow-up times were 6 years, 4 years, and 3 months. Only 1 of them had a suitable native bladder to receive the kidney graft; the other 2 required bladder augmentation, 1 of which was performed after the loss of the first graft. None of these patients had an uneventful posttransplantation course. Two patients had acute rejection episodes, and 2 had reoperations for urologic complications. One patient needed a surgical intervention owing to a sigmoid prolapse. All 3 grafts worked at last examination. The 2 patients with bladder reconstructions and longer follow-ups suffered recurrent pulmonary and urinary infections and had been hospitalized several times during each posttransplantation year. In conclusion, multiorgan involvement in VACTERL patients greatly complicates medical care after transplantation; urinary tract reconstruction seems to be essential before transplantation. PMID- 21620103 TI - Prospective study of changes in arterial stiffness among kidney-transplanted patients. AB - BACKGROUND: Chronic kidney disease is one of the main risk factors for cardiovascular disease. Changes in stiffness parameters can predict the higher risk of the development of cardiovascular disease. METHODS: Cadaveric donor kidney transplant patients (n=184) were followed in a cross-sectional single center study. Arterial stiffness parameters were measured by arteriography. We analyzed the correlation between stiffness parameters and immunosuppressive therapy, the main operation parameters, patient age, elapsed time since transplantation, carotid artery stenosis, and septual wall thickness. We enrolled 24 patients in a 3-year longitudinal study to analyze changes in stiffness parameters. RESULTS: Our cross-sectional study showed pulse wave velocity (PWV) to be significantly related to the age of the patient (P=.0001; r=0.41). There was no significant correlation between the stiffness parameters and type or dosage of immunosuppressive drugs and number of transplantations. We noted significant correlations between pulse pressure (PP) and pulse wave velocity (PWV), and augmentation index (AI) (P=.01). Patients with abnormal PWV (>12 m/s) showed significantly higher systolic blood pressures, body mass indexes, PP, and AI (P<.01). Our 3-year longitudinal study revealed a significant elevation in PWV. CONCLUSIONS: Improving endothelial function and prevention of atherosclerosis may help to reduce cardiovascular complications. Among chronic kidney disease patients, early transplantation is a possible way to prevent cardiovascular events. It is better to perform the transplantation at as early an age as possible. PMID- 21620104 TI - Functional and histopathologic changes in renal transplant patients with new onset diabetes and dyslipidemia. AB - BACKGROUND: The principal risk factors for cardiovascular mortality posttransplantation are hyperglycemia, hypertriglyceridemia, obesity, and smoking. METHODS: Among 115 patients, we assessed the risk factors for new-onset diabetes (NODM) and dyslipidemia (NODL), and their effects on the function and histopathologic changes in the allografts at 1 year posttransplantation. RESULTS: When evaluating the risk factors and the initial recipient data, we observed a significant difference in age when comparing normal vs NODM patients (P=.004), normal versus NODL patients (P=.002), and normal versus NODL + NODM patients (P=.0001). The difference in body mass index (BMI) was significant when comparing normal with NODM + NODL patients (P=.003). In regard to immunosuppressive therapy, NODM was significantly more frequent among/prescribed tacrolimus (tac; P=.005), whereas subjects who received cyclosporine (CsA) showed a significantly higher incidence of NODL (P=.001). The triglyceride levels were 3.02 +/- 1.51 mmol/L among those on CsA versus 2.15 +/- 1.57 mmol/L for (P=.004). The difference also proved to be significant for total cholesterol level: 5.43 +/- 1.23 mmol/L versus 4.42 +/- 1.31 mmol/L respectively (P=.001). In regard to allograft function a significant difference was noted at 1 year after transplantation between the NODM + NODL and the normal group in serum creatinine level (P=.02) as well as the estimated glomerular filtration rate (P=.004). Among diabetic patients, the serum creatinine level measured at posttransplant year 5 was significantly higher than that in 1 year (212.43 vs 147.00 MUmol/L; P=.0003). When assessing morphologic changes in the kidney, we observed significantly more frequent interstitial fibrosis/tubular atrophy in all 3 groups compared with normal function patients. CONCLUSION: Our clinical study suggested that at 1 year after transplantation allograft function is already impaired in the presence of both medical conditions (NODM and NODL). However, in regard to morphology, a single condition (NODM or NODL) was sufficient to produce histologic changes in the kidney. PMID- 21620105 TI - Insertion/deletion polymorphism of the angiotensin-converting enzyme predicts left ventricular hypertrophy after renal transplantation. AB - BACKGROUND: Kidney transplant recipients show a higher risk for cardiovascular complications, such as left ventricular hypertrophy and heart failure, leading to the premature death in many cases. METHODS: We investigated the contribution of angiotensin-converting enzyme (ACE) insertion/deletion (I/D) polymorphism to the development of left ventricular hypertrophy (LVH), an indicator of heart disease progression among kidney transplant recipients. RESULTS: We observed a significant correlation between graft function and left ventricular mass index. The occurrence of LVH or severe LVH was significantly greater among patients with at least one D-allele (ID or DD). CONCLUSION: The use of ACE inhibitors or angiotensin receptor blockers seemed to be advantageous for patients with the ID and especially, the DD genotype. PMID- 21620106 TI - Renal cell carcinoma of the native kidney: a frequent tumor after kidney transplantation with favorable prognosis in case of early diagnosis. AB - INTRODUCTION: The frequency of malignant tumors as a cause of death is increasing among kidney transplant patients. The aim of our study was to characterize kidney tumors occurring in the native kidneys of renal transplanted patients, and to determine their impact on recipient survival. METHODS: We retrospectively analyzed the 43/3003 (1.43%) renal cell carcinomas (RCC) in the native kidneys of patients transplanted between 1973 and 2010. RESULTS: During this period we diagnosed 293 posttransplant tumors, 14.6% of which were RCC. The male/female ratio was 2.1:1. The mean age of recipients at the time of tumor detection was 52.4 +/- 12.1 years. The mean time from transplantation to diagnosis was 72.4 +/- 61.6 months. RCC occurred on both sides in similar numbers. Tumors were multifocal in 8 cases. According to TNM staging, RCC was stage I in 38 cases. The histologic type was clear cell (n=27), papillary (n=13), chromophobe (n=2) or sarcomatoid (n=1). Radical nephrectomy was performed in 41 cases. Immunosuppressive management was converted to proliferation signal inhibitors in 27 patients (sirolimus n=19 or everolimus n=8). Fifteeen patients died at a mean survival time of 38.9 +/- 62.4 months with 28 patients still alive at a mean follow-up 43.8 +/- 35.6 months. Cumulative survival according to the Kaplan-Meier method was 79.2% at 1 year, 66.1% at 5 years, and 59.0% at 10 years. The patient survival rate was better among papillary than clear cell RCC (P=.038). CONCLUSION: RCC was the second most frequent tumor among kidney transplanted patients at our center. The diagnosis established at an early stage in the majority of cases, leading to favorable patient survivals. A regular yearly abdominal ultrasound screening is suggested for early tumor diagnosis. PMID- 21620107 TI - Examination of claudin-1 expression in patients undergoing liver transplantation owing to hepatitis C virus cirrhosis. AB - The cell adhesion molecule claudin-1 (CLDN-1) is a well known co-factor for the cell entry of hepatitis C virus (HCV). We examined 24 hepatic biopsies from liver transplant patients. Reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry were performed according to standard procedures. RT-PCR results were shown as relative expression (DeltaCT) with beta-actin as the reference gene. Immunohistochemistry results are shown by morphometry. The CLDN-1 mRNS expression rate was significantly lower when the patient displayed favorably with an unsatisfactory to antiviral therapy 0.756 +/- 0.249 versus 1.304 +/- 0.28 (P=.012). There was also a strong positive correlation between CLDN-1 protein expression and liver fibrosis (Pearson correlation coefficients: r=0.476; P=.034). PMID- 21620108 TI - The impact of Milan criteria on liver transplantation for hepatocellular carcinoma: first 15 years' experience of the Hungarian Liver Transplant Program. AB - In addition to hepatitis C, hepatocellular carcinoma. is a leading indication for orthotopic liver transplantation (OLT). The indications for OLT in HCC remains a topic of debate. The successful Milan criteria are still accepted as the gold standard to select candidates with a good chance for long-term survival. The Hungarian Liver Transplant Program launched in 1995 reached 45 OLT/year in 2010. Among 412 first OLTs, there were 49 cases of a malignant tumor, including 41 among which the indication was the tumor. Of the 412 patients, 154 (37.4%) were hepatitic C virus (HCV) positive, including 29 with HCC and 23 cases in which HCC was the indication itself. Half of the HCC patients were within the Milan criteria; 50% exceeded the criteria. We observed a solitary HCC in 36% of cases: 2 foci in 18%; 3 in 7%, 4 in 14%, and >=5 in 25%. Only 12 patients underwent a "down-staging" treatment before OLT: 8 radiofrequency ablation (RFA) and 4 transarterial chemoembolization (TACE). Cumulative 1-, 3-, and 5-year patient survivals were 62%, 54%, and 43%, respectively in HCC/HCV-positive patients and they were 74%, 67%, and 61% among non-HCC HCV-positive subjects. The cumulative HCC patient survival rates of 64%, 64%, and 53% among Milan criteria were superior to those of 57%, 40%, and 27% among subjects exceeding the Milan criteria (P=.01). Pre-OLT "down-staging" treatment increased the 1-year patient survival from 64% to 70%; however, it did not affect the long-term results. Among items of the Milan criteria tumor size had less impact on outcomes then number of foci. The majority of cases who exceeded the Milan criteria had been transplanted before 2003. Our results suggested that the Milan criteria should be applied for the selection of candidates in order to promise good survival after OLT for HCC. PMID- 21620109 TI - Volumetric hemodynamic changes and postoperative complications in hypothermic liver transplanted patients. AB - INTRODUCTION: Hepatic diseases decrease the liver's involvement in thermoregulation. Removal of the liver during transplantation increases the incidence of hypothermia during the surgery. The aims of the present study were to analyze the hemodynamic changes among hypothermic liver transplantations and to determine its relationship to postoperative complications. METHODS: Conventional and volumetric hemodynamic monitoring and intramucosal pH measurements were performed during 54 liver transplantations. According to the core temperature until graft reperfusion, patients were classified into group A, hypothermic patients (temperature < 35 degrees C; n=25) versus group B, normothermic patients (temperature > 36 degrees C; n=29). We examined the relationships between central venous pressure (CVP), intrathoracic blood volume index, cardiac index (CI), and oxygen delivery index, oxygen consumption index, as well as the fluctuation of the mean arterial pressure (MAP) and gastric intramucosal pH and activated clotting time. We recorded prolonged ventilation time, vasopressor and hemodialysis requirements, occurrence of infections, and intensive care days. RESULTS: There were no significant differences in the MELD scores. More Child-Pugh class C patients (P<.01) showed significantly higher APACHE II scores (P<.02) among group A. During hepatectomy and at the same intrathoracic blood volumes, the hypothermic group showed significantly higher CVP levels (P<.02). During the anhepatic and postreperfusion phases, the decreased CI levels (P<.05) were associated with increased MAP values (P<.05). Without differences in oxygen delivery, the oxygen consumption was lower in group A (P<.05). The intramucosal pH levels were the same in the both groups during the whole examination period. More instances of infection, intensive care, and hemodialysis treatment days, were observed as well as significantly longer vasopressor requirements and coagulopathy among the hypothermic group (P<.007). PMID- 21620110 TI - Liver transplantation for acute liver failure: the Hungarian experience. AB - Acute liver failure (ALF) counts for 9%-11% of activity in leading liver transplant programs. We have summarized the Hungarian Liver Transplant Program experience for ALF among 412 consecutive orthotopic liver transplantations (OLTs). All OLTs were performed without an extended international donor background. The proportion of ALF among the indications for OLT was lower (5.8% vs 9%) and early mortality higher than the European Liver Transplant Registry (1 year cumulative patients survival is 70% in ELTR vs 60% in the HU LT Program). The waiting time for a donor was longer than expected in the Eurotransplant community. Regarding postoperative complications, there was a higher incidence of initial poor function, bacterial infection, sepsis, and multiorgan failure. We conclude that ALF can be managed with reasonable results but requires an extended donor pool with an integrated international network to improve postoperative morbidity and mortality. PMID- 21620111 TI - Relationship between hepatitis C virus recurrence and de novo diabetes after liver transplantation: the Hungarian experience. AB - De novo diabetes mellitus is a common complication after liver transplantation. It is strongly associated with hepatitis C virus (HCV) infection. We analyzed the relationship between HCV recurrence and de novo diabetes among the Hungarian liver transplant population. This retrospective study included cases from 1995 to 2009 on 310 whole liver transplantations. De novo diabetes was defined if the patient had a fasting plasma glucose >=126 mg/dL permanently after the third month post liver transplantation, and/or required sustained antidiabetic therapy. De novo diabetes occured in 63 patients (20%). The cumulative patient survival rates at 1, 3, 5, and 8 years were 95%, 91%, 88%, and 88% in the control group, and 87%, 79%, 79%, and 64% in the de novo group, respectively (P=.011). The majority of the patients in the de novo group were HCV positive (66% vs 23%). Early virus recurrence within 5 months was associated with the development of diabetes (80% vs 20% non-diabetic controls; P=.017). The fibrosis (2.05 +/- 1.5 vs 1 +/- 1; P=.039) and Knodell scores (3.25 +/- 2 vs 1.69 +/- 1.2; P=.019) were higher among the de novo group after antiviral therapy. Rapid recurrence, more severe viremia, and fibrosis showed significant roles in the developement of de novo diabetes after liver transplantation. PMID- 21620112 TI - Patchy myocardial pattern of virus sequence persistence in heart transplant recipients--possible role of sampling error in the etiology. AB - BACKGROUND: The pathway from viral myocarditis to end-stage heart failure is commonly accepted, but diagnosis of virus-mediated myocardial injury remains challenging. Virus persistency in the myocardium may accelerate ventricular failure; thus, a precise diagnosis of virus persistency may prevent the development of end-stage heart failure. METHODS: We performed a systematic investigation on the sampling error of viral diagnostics in heart transplant recipients: Transmural samples from 5 regions of the explanted hearts from recipients during heart transplantation were amplified using entero-, adeno-, and herpesvirus sequences and histologic examinations performed. RESULTS: We examined 175 myocardial samples from dilated cardiomyopathy and 100 samples from 20 forensic medicine patients. Seven patients were positive for the examined viruses: 10 positive regions for adenovirus, and 1 positive region for herpes virus DNA, but none for enterovirus. A focal myocardial pattern was detected for adenovirus. CONCLUSION: Our results with the patchy myocardial viral persistence may explain possible false-negative results related to virus-mediated etiology among end-stage dilated cardiomyopathy patients. Therefore, repeated endomyocardal biopsies, and multiple cardiac samples are recommended to be obtained to evaluate the etiology of heart failure, thus reducing the occurrence of end-stage heart failure and decreasing the number of patients requiring heart transplantation. PMID- 21620113 TI - Airway pathogens during the first year after lung transplantation: a single center experience. AB - After lung transplantation, a high level of immunosuppression is needed to prevent rejection. This demand renders recipients more sensitive to infections. As pulmonary infections are a major clinical problem during the first postoperative year after lung transplantation, preventive treatment and regular surveillance examinations are needed for immediate, adequate therapy. We describe the airway pathogens registered during the first posttransplantation year among our 12 lung transplant recipients since December 2008. Samples were obtained for microbiologic analysis from the upper and lower respiratory tracts and from serum as part of routine care. During the first year after transplantation the most frequent pathogens were fungi (Candida albicans 82%; Aspergillus 50%), Pneumocystis (8%), gram-negative bacteria (Pseudomonas spp 60%; Klebsiella 25%, Acinetobacter 17%; Escherichia Coli 17%; and Enterococcus faecalis 25%), and Staphylococcus aureus (50%, including methicillin-resistant strains 25%). This pathogen spectrum in the first postoperative year after lung transplantation was similar to other centers. Colonization with Pseudomonas or fungi presented early and was prevalent among our patients. PMID- 21620114 TI - Severe mixed sleep apnea after bilateral lung transplantation in a cystic fibrosis patient: a case report. AB - BACKGROUND: Lung transplantation is the only treatment for end-stage lung disease in selected patients. After lung transplantation, patient recovery is often slow owing to severe underlying diseases in the patient producing hypoxemia before, during, and after surgery, as well as infections and rejection episodes. Postoperative breathing and ventillatory disorders are also associated with diaphragmatic dysfunction and/or phrenic nerve damage. METHODS: Herein we have reported a case of a 35-year-old man undergoing bilateral lung transplantation owing to worsening of chronic respiratory failure from cystic fibrosis. After uncomplicated surgery, weaning was delayed due to nighttime dyspnea and hypoxemia attributed to diaphragm dysfunction. After improvement of diaphragm function, the symptoms persisted, requiring noninvasive nocturnal ventilatory support. Polysomnography confirmed severe mixed sleep apnea. RESULTS: Effective treatment with noninvasive bi-level positive airway pressure spontaneous/timed mode (BiPAP S/T) ventilation during the nights rendered the patient symptom free. Polysomnography confirmed successful treatment. CONCLUSION: Disordered breathing while sleeping is common after solid organ transplantation. BiPAP S/T ventilator therapy was effective to the treat dominantly central sleep apnea in our patient. PMID- 21620115 TI - Presence of pituitary adenylate cyclase activating polypeptide and its type I receptor in the rat kidney. AB - Pituitary adenylate cyclase activating polypeptide (PACAP), a multifunctional neuropeptide, has 2 active forms, PACAP38 and PACAP27. It is now well-established that PACAP has several actions also in peripheral organs, including renoprotective effects. The peptide itself has not been previously identified in the rat kidney. The first aim of our study was to identify PACAP in the rat kidney using mass spectrometry and radioimmunoassay (RIA). Receptor mRNA and binding studies revealed the existence of all 3 PACAP receptors (PAC1, VPAC1, and VPAC2) in the kidney, but their exact localization in histologic sections was not evident. Because most of the cytoprotective effects of PACAP relate to its specific PAC1 receptor, our second aim was to identify the cell types wherein the PAC1 receptor is expressed in the rat kidney. Mass spectrometry revealed the presence of PACAP38 in the kidney. RIA measurements showed both PACAP38- and PACAP27-like immunoreactivities in kidney homogenates, with PACAP38 being dominant. Immunohistochemistry revealed PAC1 receptor-like immunoreactivity in kidney sections, mainly expressed in cortical tubular epithelial cells. These results showed PACAP to be endogenously present in the kidney. The tubular localization of the PAC1 receptor provides the basis for the renal effects of the peptide under physiologic and pathologic conditions. PMID- 21620116 TI - Incisional hernia after simultaneous pancreas kidney tranplantation: a single center experience from Budapest. AB - INTRODUCTION: The occurrence of postoperative incisional hernia is more frequent after simultaneous pancreas-kidney transplantation compared with other transplanted parenchymal organs. These complications are especially dangerous in this patient population, because they can compromise the survival of the transplanted organ. METHODS: We performed a retrospective review of a series of adult patients with incisional herniae after 23 consecutive simultaneous pancreas kidney transplantations between January 2004 and June 2010 seeking to identify risk factors. All 23 patients had a body mass index (BMI) of <25. All surgeons used a similar technique, including a median incision with an intraperitoneal approach, and systemic venous and enteric drainage methods and a layered fascial closure. All combined pancreas-kidney transplant recipients received induction with thymoglobulin and maintenance therapy with sirolimus, reduced-dose cyclosporine and corticosteroids. RESULTS: An incisional hernia repair was performed in 8/23 patients (34.8%). Four reoperations were required in this group (50%), due to hemoperitoneum (n=2), intra-abdominal abscess (n=1), and venous thrombosis (n=1). The mean elapsed time between transplantation and hernioplasty was 24.5 months (range, 8-51). There was no significant difference in age, gender, BMI, dialysis modality, or operative time among affected compared with the other members of the group. CONCLUSION: Despite lack of obesity we observed a relatively higher rate of postoperative herniase, possibly owing to the side effects of a thymoglobulin-sirolimus combination. PMID- 21620117 TI - Organ donation and transplantation from medical students' perspective: introducing the experience from an academic league in Brazil. AB - INTRODUCTION: Medical education is an important factor to decrease the waiting list for transplantation. Reports in the medical literature reveal limited notification of potential organ donors by general physicians. Appropriate information is also needed to increase the availability of potential donors and minimize the waiting list. This article describes the acquired experience with an extracurricular program of education on organ and tissue transplantation in our institution, searching to meet medical information needs using a format of an academic league. METHODS: This qualitative study describes a proposed approach on the theme of "transplant and organ and tissue donation" with medical students from a Brazilian university, through creation of a program named "Transplantation League" in direct association with a transplantation center. RESULTS: The league's activities are based on three main activities teaching, research, and practical. Besides the organization of the I Course of Organ and Tissue Transplantation, the project received financial support from the Federal University of Goias to develop the assignments. A member's stake in the league included scientific projects involving liver transplantation candidates, as well as notification, donation, transportation, and transplantation of these patients. CONCLUSION: The academic league has the purpose of academic information on organ and tissues transplantation. Its application in medical schools may be valuable to increase transplant numbers. PMID- 21620118 TI - Perfusion fluid contamination in relation to recipient survival and acute cellular rejection in orthotopic liver transplantation: retrospective analysis. AB - INTRODUCTION: A perfusion fluid used in the preservation of a grafted liver represents a medium suitable for microorganism growth. This study investigated the prevalence of perfusion fluid contamination, acute cellular rejection (ACR) episodes, and patient survival rate. METHOD: This is a retrospective study, based on an electronic database allocating cases of orthotopic liver transplantation. The exclusion criteria were as follows: having been submitted to multiple organ transplantation, liver retransplantation only, and those whose samples had not been collected or sent on the back table procedure or were unobtainable (usually the samples were sent when there was donor infection suspicion/positivity). Our posttransplantation infection prophylactic protocol consisted of ampicillin/sulbactam for 72 hours. The variables in the study were as follows: fluid contamination, presence of acute cellular rejection (ACR, Banff classification), and recipient survival at the first year. Statistical analysis was performed using descriptive statistics and chi-square with Fisher exact test considering significant P<.05. RESULTS: We observed perfusion fluid contamination in 15/121 (12.39%). The agents were as follows: Klebsiella pneumoniae in 6 (4.96%), Staphylococcus epidermidis in 5 (4.13%), and Acinetobacter baumanii in 3 (2.48%) and negative cultures in 106 (87.60%). Only 1 patient had matching for donor infection and positivity hemoculture after the transplantation (K pneumoniae) and he was the only patient associated with fluid infection and death. The recipients who had their fluid preservation with positive cultures had more ACR and the survival rate was similar among those with or without infection. CONCLUSION: Optimization of microbiological procedures can be performed including fungal and bacterial cultures. PMID- 21620119 TI - Oral health profile of cirrhotic patients awaiting liver transplantation in the Brazilian Northeast. AB - BACKGROUND: Infections are a frequent cause of morbidity and mortality among postoperative liver transplant (OLT) patients and a leading cause of decompensated chronic liver disease (CLD) among patients awaiting the procedure. Oral lesions that are frequently observed in subjects with CLD may represent foci for systemic infections before and after OLT. AIMS: To evaluate the oral health profile of patients with CLD awaiting OLT. METHODS: One hundred thirty one patients including 100 males of overall mean age 49.5 +/- 10.8 years with CLD were listed for OLT and examined for oral health status according to a established protocol. RESULTS: One hundred thirty (99%) patients were partially edentulous; 66 (51%) had chewing difficulties; and 63 (48%) experienced reduced salivary flow. With respect to periodontal disease and oral infections, 68 (25%) had periodontitis, 63 (48%) had periapical lesion, 64 (49%) had abscesses, and 59 (45%) had root fragments. Loss of follow-up was observed in 21 subjects. Among the 110 other patients, 63 (57%) underwent dental treatments with complications in only two cases. Interestingly, mortality was significantly lower among treated (31%) versus nontreated patient (79%; P<.001). CONCLUSIONS: Poor oral health status observed in most CLD patients may represent a source of systemic infections before and after OLT. Treatment of such lesions was feasible in the majority of the patients and seemed to be associated with a reduction in mortality. PMID- 21620120 TI - Validation of 20-meter corridor for the 6-minute walk test in men on liver transplantation waiting list. AB - BACKGROUND: Guidelines established by the American Thoracic Society recommend the use of corridors 30 m in length for the 6-minute walk test (6MWT). However, not all institutions have such long corridors, which hinders or prevents 6MWT performance and use of its benefits. OBJECTIVE: To compare the distances walked by male patients with cirrhosis on the liver transplantation waiting list, we performed 6MWT on corridors 20 and 30 m long. METHODS: This prospective study included 10 patients on the waiting list for liver transplantation. They underwent 2 walk tests: the first test in a 20 m corridor and the second in a 30 m corridor. We assessed physiologic variables (heart rate, oxygen saturation, arterial blood pressure) and the subjective sensation of dyspnea at rest at 6 and 9 minutes after each walk. Statistical analysis was performed using the Wilcoxon test. RESULTS: Data are expressed as mean +/- sd. Patient age was 59 +/- 10 years. The distances walked by the patients were shorter than those predicted for their age and gender (586 +/- 45 m) in both tests, no significant difference was observed between the first and the second 6MWT (437 +/- 101 m vs 465 +/- 80 m; P=.131) or among the physiologic variables. CONCLUSION: We concluded that a 20 m corridor can be used safely and effectively as an alternative to 30 m for the 6MWT for male patients with cirrhosis on the liver transplantation waiting list. PMID- 21620121 TI - Comparison of surface electromyography in respiratory muscles of healthy and liver disease patients: preliminary studies. AB - INTRODUCTION: Surface electromyography (SEMG) is described as a technique to detect voluntary muscle activity. In the respiratory muscles, diaphragm and the rectus abdominis-are especially important for respiratory compliance. OBJECTIVE: We sought to study the activity of muscles using SEMG of the right diaphragm and the right rectus abdominis in healthy subjects versus liver disease patients (LDS). METHOD: Each group of 30 male patients underwent SEMG with electrodes attached to the dermis surface at the xiphoid and below the right costal margin (channel 1). For the rectus abdominis, we placed the electrodes on the right 5 cm below the umbilicus. The variables studied were: root mean square (RMS), maximum inspiratory pressure (MIP), and maximum expiratory pressure (MEP). We also evaluated age, weight, body mass index, smoking history lifestyle sedentary preexistent chronic lung disease. Nonparametric tests were used for statistical analysis. RESULTS: There were significant differences (P>0001) between the groups regarding MEP and MIP values showing low pressures in the LDS group. RMS of the rectus abdominis showed a trend (P=.059) toward compliance in the LDS group. Compared with the healthy subjects, there were 18% versus 5% sedentary individuals; only 10% versus 27.3% were smokers or former smokers and chronic lung disease was present in 2% versus 4%. CONCLUSION: The respiratory muscle evaluation using SEMG detected decreased respiratory muscle strength and a trend to rectus abdominis compliance among LDS. PMID- 21620122 TI - Orthotopic liver transplantation without venovenous bypass using the conventional and piggyback techniques. AB - INTRODUCTION: Orthotopic liver transplantation is a widely used procedure for the treatment of irreversible liver diseases for which there is no possibility of medical treatment. When this procedure is performed by the conventional technique, the retrohepatic vena cava is removed along with the native liver. The inferior vena cava (IVC) remains clamped until the revascularization of the graft, and in this period there is a reduction in the venous return, which may induce a fall by up to 50% in the cardiac output with hemodynamic instability and a fall in renal perfusion pressure. The use of a portal-femoral-axillary venovenous bypass system, in which the blood from the femoral and portal veins returns to the heart via the axillary vein propelled by a centrifugal pump, is intended to minimize the effects of the IVC clamping. In the piggyback (PB) technique, the native liver is removed and the IVC of the recipient is preserved and only partially clamped. We have employed both techniques without the use of venovenous bypass for 10 years. The objective of this study was to compare the results obtained from the use of the two techniques. PATIENTS AND METHODS: A retrospective analysis was performed of 195 patients transplanted between 1999 and 2008: 125 by the conventional technique and 70, the PB technique. The intraoperative parameters were analyzed (surgical time, ischemia time, use of blood products, and diuresis), as well as intensive care support (duration of stay in intensive care unit and use of vasoactive drugs), period of intubation, length of hospital stay, renal function, graft function, postoperative complications, retransplantation, and patient survival. RESULTS: The PB group showed a reduction in surgical time, warm ischemia time, the use of packed red blood cells concentrates, and fresh frozen plasma, as well as mortality at 30 days (P<.05). There were no differences in relation to cold ischemia time, intraoperative diuresis; length of stay and use of vasoactive drugs in the intensive care unit; the period of intubation; the duration of hospital stay; the renal function; the graft function; the need for reoperation; the incidence of sepsis, biliary complications, vascular complications; need for retransplantation; and 1-year mortality. The cumulative survival rate at 1 year was significantly better among the PB patients. CONCLUSION: Orthotopic liver transplantation can be performed without venovenous bypass with good results, using either the conventional technique or the PB technique. Provided that there is no technical contraindication and a long ischemia period is not foreseen, the PB technique should be the technique of choice. PMID- 21620123 TI - Pregnancy outcome in female liver transplant recipients. AB - Transplantation has become an available and successful treatment option for numerous congenital and acquired hepatic disorders. Studies have shown that when the prepregnancy recipient graft function is stable and adequate, pregnancy is normally well tolerated with favorable neonatal outcomes. However, there are reports of increased incidences of hypertension and preeclampsia as well as lower birth weights and prematurity. Patients administered tacrolimus-based therapies seem to have lower incidences of these complications. CASE REPORTS: The 5 reported patients, aged 23-37 years at the time of conception, were 2-11 years posttransplantation. A preterm delivery for fetal distress was the most clinically important complication among these patients. One episode of acute genital herpes infection, 1 liver hematoma in a patient who was anticoagulated owing to a history of deep vein thrombosis, and 1 case of wound infection postpartum were also observed. Despite these complications, all 5 pregnancies were successful. The mean gestational age at delivery was 35.2 weeks. No structural malformations or early complications were observed in the neonates. All cases showed stable liver parameters. PMID- 21620124 TI - Application of the intelligent techniques in transplantation databases: a review of articles published in 2009 and 2010. AB - The replacement of defective organs with healthy ones is an old problem, but only a few years ago was this issue put into practice. Improvements in the whole transplantation process have been increasingly important in clinical practice. In this context are clinical decision support systems (CDSSs), which have reflected a significant amount of work to use mathematical and intelligent techniques. The aim of this article was to present consideration of intelligent techniques used in recent years (2009 and 2010) to analyze organ transplant databases. To this end, we performed a search of the PubMed and Institute for Scientific Information (ISI) Web of Knowledge databases to find articles published in 2009 and 2010 about intelligent techniques applied to transplantation databases. Among 69 retrieved articles, we chose according to inclusion and exclusion criteria. The main techniques were: Artificial Neural Networks (ANN), Logistic Regression (LR), Decision Trees (DT), Markov Models (MM), and Bayesian Networks (BN). Most articles used ANN. Some publications described comparisons between techniques or the use of various techniques together. The use of intelligent techniques to extract knowledge from databases of healthcare is increasingly common. Although authors preferred to use ANN, statistical techniques were equally effective for this enterprise. PMID- 21620125 TI - Artificial intelligence techniques: predicting necessity for biopsy in renal transplant recipients suspected of acute cellular rejection or nephrotoxicity. AB - The gold standard for nephrotoxicity and acute cellular rejection (ACR) is a biopsy, an invasive and expensive procedure. More efficient strategies to screen patients for biopsy are important from the clinical and financial points of view. The aim of this study was to evaluate various artificial intelligence techniques to screen for the need for a biopsy among patients suspected of nephrotoxicity or ACR during the first year after renal transplantation. We used classifiers like artificial neural networks (ANN), support vector machines (SVM), and Bayesian inference (BI) to indicate if the clinical course of the event suggestive of the need for a biopsy. Each classifier was evaluated by values of sensitivity and area under the ROC curve (AUC) for each of the classifiers. The technique that showed the best sensitivity value as an indicator for biopsy was SVM with an AUC of 0.79 and an accuracy rate of 79.86%. The results were better than those described in previous works. The accuracy for an indication of biopsy screening was efficient enough to become useful in clinical practice. PMID- 21620126 TI - Non-human leukocyte antigen antibodies reactive with endothelial cells could be involved in early loss of renal allografts. AB - Preformed donor-specific human leukocyte antigen (HLA) antibodies have been associated with allograft dysfunction and failure. However, recipients of HLA identical kidneys can develop acute humoral rejection, implicating putative pathogenic antibodies that are directed against non-HLA antigens. We investigated the presence of endothelial cell-reactive antibodies in 11 patients who experienced early loss of their transplanted kidneys owing to humoral rejection and 1 loss from renal venal thrombosis. We examined the potential efficacy of intravenous immunoglobulin to block the binding of these antibodies, as previously suggested for anti-HLA antibodies. PMID- 21620127 TI - Decrease in bone mass in women after liver transplantation: associated factors. AB - BACKGROUND: In the future, an increasing number of female liver transplant recipients will reach the climacteric with osteoporosis as a common complication. We evaluated the factors associated with decreased bone mass among women after liver transplantation. METHODS: A prospective, cross-sectional study of 23 outpatient transplant recipients followed from February 2009 to March 2010 included women of age >=35 years after liver transplantations >=1 year prior. We recorded patient histories, liver enzyme levels, as well as bone mineral densities measured at the lumbar spine and femur. Statistical analysis used Fisher's exact test, simple odds ratio (OR), and Spearman's rank correlation coefficient. RESULTS: The mean patient age was 52.5 +/- 11 years with 30.4% premenopausal, and 69.6% perimenopausal or postmenopausal. Approximately 21% showed osteoporosis and 35%, a low bone mass. Postmenopausal women: OR 69.0 (95% CI 2.89-1647.18; P<.0001), aged >=49 years: OR 13.33 (95% CI 1.78-100.15; P=.0123) and receiving a transplant after 44 years of age: OR 49.50 (95% CI 3.84 638.43; P<.0001) were associated with a lower bone mass. Having undergone transplantation for more than 5.8 years lowered the risk of bone mass change: OR 0.11 (95% CI 0.02-0.78; P=.0361). Clinical and laboratory variables, including corticosteroid use, were not associated with decreased bone mass. CONCLUSION: Understanding the prevalence and factors associated with osteoporosis among female liver transplant recipients is important to enhance the strategies to diagnose and treat these women, seeking to improve their quality of life. PMID- 21620128 TI - Cytomegalovirus, human herpesvirus-6, and human herpesvirus-7 in adult liver transplant recipients: diagnosis based on antigenemia. AB - Human herpesvirus (HHV)-6, HHV-7, and cytomegalovirus (CMV) that remain latent after primary infection can be reactivated during immunosuppression following organ transplantation in liver transplant recipients. The aim of this study was to monitor active infections for HHV-6, HHV-7, and CMV among adult liver transplantation recipients using antigenemia detected by an immunoperoxidase staining. Twenty-eight adult liver transplant patients were monitored using antigenemia in blood samples obtained at the time of transplantation, as well as weekly in the first month and once a month for 6 months. Of these patients, 28.5% showed positive CMV antigenemia; 39.2%, HHV-6 antigenemia; and 14.2%, HHV-7 antigenemia. The detection of the three viruses was considered to be independent of one another (P>.05). The results described above showed that few patients remain free of beta herpesviruses after liver transplantation. Most patients were infected sequentially and not concurrently. Antigenemia has been considered useful to detect active HHV-6 and HHV-7 infections. Antigenemia can be more efficiently interpreted when compared with polymerase chain reaction results, although other studies are necessary to establish the reference of HHV-6 and HHV 7 antigenemia. PMID- 21620129 TI - Monitoring and detection of cytomegalovirus in liver transplant recipients. AB - Cytomegalovirus (CMV) is a beta-herpesvirus. CMV infections are a common complication contributing to morbidity and mortality after liver transplantation. Among organ transplant recipients, CMV can reactivate from latency during the first 6 months. This prospective study performed from February 2008 to December 2009 examined liver transplant recipients during the first 6 months. Two methods were performed to detect CMV infections: antigenemia (AGM) and nested (PCR). Ninety-four patients, including 72 men (76.6%) and 22 women (23.4%) underwent liver transplantation during this period. We analyzed 575 samples including 465 for AGM and PCR. Forty-three (9.25%) showed positive AGM as detected 2 to 179 days posttransplantation with a mean of 50 days and a median of 35 days, and 93/465 (20%) showed positive PCR at 0 to 186 days posttransplantation with a mean of 31 days and a median of 38 days. Among the 43 antigenemia patients, 38 samples were positive for up to 5 cells 18 of which were PCR-positive. Five samples were positive with more than 5 cells, including 3 that were PCR-positive. Only 4.51% had AGM and were PCR-positive in the same sample. Despite only 9.25% (43/465) showing AGM, the current study suggested the utility of routine monitoring to detect early CMV infection among liver transplantation patients seeking to reduce morbidity and mortality. PMID- 21620130 TI - Prognostic factors for hepatocellular carcinoma recurrence: experience with 83 liver transplantation patients. AB - INTRODUCTION: Orthotopic liver transplantation (OLT) is a rational therapeutic option for early-stage hepatocellular carcinoma (HCC) providing a potential cure and improving survival. METHODS: This retrospective study of a longitudinal cohort used an electronic database collected prospectively from September 1997 to May 2010. The variables were gender, age (years), and alpha-fetoprotein (AFP) level (ng/mL). In explanted livers we observed: microvascular or macrovascular invasion, number of nodules and their largest size, Edmondson-Steiner histological differentiation, incidental tumor transarterial chemoembolization (TACE), Milan criteria, and previous down-staging. RESULTS: Five of 83 (6.0%) subjects including 68 (82%) males with a mean time to diagnosis of 9 months experienced tumor relapses. Mean patient age at HCC recurrence was 55.3 years for male and 44.6 years for female subjects. Vascular invasion was detected in 17/83 (20.5%) subjects, namely 2% of macrovascular invasion, and 52.5% with expanded Milan criteria due to an increased number and size of nodules in the explanted livers. An incidental tumor was observed in 29.5% of cases. Preoperative TACE treatment was performed in 13 (15.6%) patients. None of the patients who had a HCC recurrence had undergone TACE. AFP level at the time of recurrence was around 1,900 ng/mL. The predictive factor for mortality was nodule size (P=.04; hazard ratio=0.0269; confidence interval [CI], 95% 0.0094-0.299). CONCLUSION: Patients with relapses showed the worst survival and tumor size was a predictive factor for recurrence. PMID- 21620131 TI - Taking Charge: Feeding Malaria via Anion Channels. AB - The human malaria parasite Plasmodium falciparum increases red blood cell membrane permeability during infection to allow for import of nutrients and other solutes. Nguitragool et al. (2011) have now identified parasite-encoded CLAG3 proteins as key components of the import channel located on the erythrocyte membrane. PMID- 21620132 TI - PK-M2 Makes Cells Sweeter on HIF1. AB - The transcription factor hypoxia-inducible factor 1 (HIF1) facilitates the induction of enzymes necessary for anaerobic glycolysis. Luo et al. (2011) now identify pyruvate kinase (PK)-M2 as an intriguing new interacting partner for HIF1, revealing a potential mechanism for the Warburg effect, an elevation in aerobic glycolytic metabolism frequently observed in cancer. PMID- 21620133 TI - Vertebrate segmentation: from cyclic gene networks to scoliosis. AB - One of the most striking features of the human vertebral column is its periodic organization along the anterior-posterior axis. This pattern is established when segments of vertebrates, called somites, bud off at a defined pace from the anterior tip of the embryo's presomitic mesoderm (PSM). To trigger this rhythmic production of somites, three major signaling pathways--Notch, Wnt/beta-catenin, and fibroblast growth factor (FGF)--integrate into a molecular network that generates a traveling wave of gene expression along the embryonic axis, called the "segmentation clock." Recent systems approaches have begun identifying specific signaling circuits within the network that set the pace of the oscillations, synchronize gene expression cycles in neighboring cells, and contribute to the robustness and bilateral symmetry of somite formation. These findings establish a new model for vertebrate segmentation and provide a conceptual framework to explain human diseases of the spine, such as congenital scoliosis. PMID- 21620134 TI - Malaria parasite clag3 genes determine channel-mediated nutrient uptake by infected red blood cells. AB - Development of malaria parasites within vertebrate erythrocytes requires nutrient uptake at the host cell membrane. The plasmodial surface anion channel (PSAC) mediates this transport and is an antimalarial target, but its molecular basis is unknown. We report a parasite gene family responsible for PSAC activity. We used high-throughput screening for nutrient uptake inhibitors to identify a compound highly specific for channels from the Dd2 line of the human pathogen P. falciparum. Inheritance of this compound's affinity in a Dd2 * HB3 genetic cross maps to a single parasite locus on chromosome 3. DNA transfection and in vitro selections indicate that PSAC-inhibitor interactions are encoded by two clag3 genes previously assumed to function in cytoadherence. These genes are conserved in plasmodia, exhibit expression switching, and encode an integral protein on the host membrane, as predicted by functional studies. This protein increases host cell permeability to diverse solutes. PMID- 21620135 TI - DNA replication through G-quadruplex motifs is promoted by the Saccharomyces cerevisiae Pif1 DNA helicase. AB - G-quadruplex (G4) DNA structures are extremely stable four-stranded secondary structures held together by noncanonical G-G base pairs. Genome-wide chromatin immunoprecipitation was used to determine the in vivo binding sites of the multifunctional Saccharomyces cerevisiae Pif1 DNA helicase, a potent unwinder of G4 structures in vitro. G4 motifs were a significant subset of the high confidence Pif1-binding sites. Replication slowed in the vicinity of these motifs, and they were prone to breakage in Pif1-deficient cells, whereas non-G4 Pif1-binding sites did not show this behavior. Introducing many copies of G4 motifs caused slow growth in replication-stressed Pif1-deficient cells, which was relieved by spontaneous mutations that eliminated their ability to form G4 structures, bind Pif1, slow DNA replication, and stimulate DNA breakage. These data suggest that G4 structures form in vivo and that they are resolved by Pif1 to prevent replication fork stalling and DNA breakage. PMID- 21620136 TI - Maternal epigenetic pathways control parental contributions to Arabidopsis early embryogenesis. AB - Defining the contributions and interactions of paternal and maternal genomes during embryo development is critical to understand the fundamental processes involved in hybrid vigor, hybrid sterility, and reproductive isolation. To determine the parental contributions and their regulation during Arabidopsis embryogenesis, we combined deep-sequencing-based RNA profiling and genetic analyses. At the 2-4 cell stage there is a strong, genome-wide dominance of maternal transcripts, although transcripts are contributed by both parental genomes. At the globular stage the relative paternal contribution is higher, largely due to a gradual activation of the paternal genome. We identified two antagonistic maternal pathways that control these parental contributions. Paternal alleles are initially downregulated by the chromatin siRNA pathway, linked to DNA and histone methylation, whereas transcriptional activation requires maternal activity of the histone chaperone complex CAF1. Our results define maternal epigenetic pathways controlling the parental contributions in plant embryos, which are distinct from those regulating genomic imprinting. PMID- 21620137 TI - An intercellular heme-trafficking protein delivers maternal heme to the embryo during development in C. elegans. AB - Extracellular free heme can intercalate into membranes and promote damage to cellular macromolecules. Thus it is likely that specific intercellular pathways exist for the directed transport, trafficking, and delivery of heme to cellular destinations, although none have been found to date. Here we show that Caenorhabditis elegans HRG-3 is required for the delivery of maternal heme to developing embryos. HRG-3 binds heme and is exclusively secreted by maternal intestinal cells into the interstitial fluid for transport of heme to extraintestinal cells, including oocytes. HRG-3 deficiency results either in death during embryogenesis or in developmental arrest immediately post-hatching phenotypes that are fully suppressed by maternal but not zygotic hrg-3 expression. Our results establish a role for HRG-3 as an intercellular heme trafficking protein. PMID- 21620138 TI - Pyruvate kinase M2 is a PHD3-stimulated coactivator for hypoxia-inducible factor 1. AB - The pyruvate kinase isoforms PKM1 and PKM2 are alternatively spliced products of the PKM2 gene. PKM2, but not PKM1, alters glucose metabolism in cancer cells and contributes to tumorigenesis by mechanisms that are not explained by its known biochemical activity. We show that PKM2 gene transcription is activated by hypoxia-inducible factor 1 (HIF-1). PKM2 interacts directly with the HIF-1alpha subunit and promotes transactivation of HIF-1 target genes by enhancing HIF-1 binding and p300 recruitment to hypoxia response elements, whereas PKM1 fails to regulate HIF-1 activity. Interaction of PKM2 with prolyl hydroxylase 3 (PHD3) enhances PKM2 binding to HIF-1alpha and PKM2 coactivator function. Mass spectrometry and anti-hydroxyproline antibody assays demonstrate PKM2 hydroxylation on proline-403/408. PHD3 knockdown inhibits PKM2 coactivator function, reduces glucose uptake and lactate production, and increases O(2) consumption in cancer cells. Thus, PKM2 participates in a positive feedback loop that promotes HIF-1 transactivation and reprograms glucose metabolism in cancer cells. PMID- 21620139 TI - Primate CpG islands are maintained by heterogeneous evolutionary regimes involving minimal selection. AB - Mammalian CpG islands are key epigenomic elements that were first characterized experimentally as genomic fractions with low levels of DNA methylation. Currently, CpG islands are defined based on their genomic sequences alone. Here, we develop evolutionary models to show that several distinct evolutionary processes generate and maintain CpG islands. One central evolutionary regime resulting in enriched CpG content is driven by low levels of DNA methylation and consequentially low rates of CpG deamination. Another major force forming CpG islands is biased gene conversion that stabilizes constitutively methylated CpG islands by balancing rapid deamination with CpG fixation. Importantly, evolutionary analysis and population genetics data suggest that selection for high CpG content is not a significant factor contributing to conservation of CpGs in differentially methylated regions. The heterogeneous, but not selective, origins of CpG islands have direct implications for the understanding of DNA methylation patterns in healthy and diseased cells. PMID- 21620141 TI - SnapShot: the segmentation clock. PMID- 21620140 TI - Analysis of the human endogenous coregulator complexome. AB - Elucidation of endogenous cellular protein-protein interactions and their networks is most desirable for biological studies. Here we report our study of endogenous human coregulator protein complex networks obtained from integrative mass spectrometry-based analysis of 3290 affinity purifications. By preserving weak protein interactions during complex isolation and utilizing high levels of reciprocity in the large dataset, we identified many unreported protein associations, such as a transcriptional network formed by ZMYND8, ZNF687, and ZNF592. Furthermore, our work revealed a tiered interplay within networks that share common proteins, providing a conceptual organization of a cellular proteome composed of minimal endogenous modules (MEMOs), complex isoforms (uniCOREs), and regulatory complex-complex interaction networks (CCIs). This resource will effectively fill a void in linking correlative genomic studies with an understanding of transcriptional regulatory protein functions within the proteome for formulation and testing of future hypotheses. PMID- 21620142 TI - Minimizing obstacles and maximizing outcomes in microvascular breast reconstruction. PMID- 21620143 TI - Overview of perforator imaging and flap perfusion technologies. AB - Breast reconstruction has become an important consideration for women after mastectomy. Over the past decade, there have been a variety of technological advancements that have facilitated the ability to deliver reproducible and predictable outcomes with autologous breast reconstruction. This article chronicles many of the technological advancements and reviews the current toolbox that surgeons now have at their disposal when performing autologous reconstruction. It focuses on preoperative, intraoperative, and postoperative tools that have enabled the achievement of more reliable and predictable outcomes, especially in the setting of microvascular breast reconstruction. PMID- 21620144 TI - Assessing perforator architecture. AB - A myriad of options exist for autologous tissue perforator-based breast reconstruction. Each perforator selected has its distinct vascular territory, and proper knowledge of perforator perfusion characteristics helps maximize outcome and limit complications. PMID- 21620145 TI - Acoustic Doppler sonography, color duplex ultrasound, and laser Doppler flowmetry as tools for successful autologous breast reconstruction. AB - The sine qua non to best ensure viability of any autogenous tissues used for breast reconstruction is to maximize the appropriate circulatory pattern to that tissue. This overview of tools used in this regard, all based on the physical principles of the Doppler effect, compares the role today of acoustic Doppler sonography, color duplex ultrasound, and laser Doppler flowmetry for perforator identification and flap monitoring. The audible Doppler has recognized limitations, but remains the simplest and most universally available device to assist in this purpose. Laser Doppler flowmetry provides a reasonable system for both intraoperative and post-procedure objective monitoring of the chosen tissue transfer. PMID- 21620146 TI - Maximizing the use of the handheld Doppler in autologous breast reconstruction. AB - The handheld acoustic Doppler is commonly used in the setting of autologous reconstruction. Like the color duplex Doppler, it can be used preoperatively but has the advantage of being used intraoperatively and postoperatively. It can be used as a sole modality or it can be used in conjunction with or to confirm the findings of CTA or MRA. This article focuses on the preoperative, intraoperative, and postoperative use of the handheld Doppler for free tissue transfer with an emphasis on perforator flap breast reconstruction. PMID- 21620147 TI - Computerized tomographic and magnetic resonance angiography for perforator-based free flaps: technical considerations. AB - Perforator-based free flaps rely on the appropriate dominant vessel supplying the vascular territory of the flap. Preoperative knowledge of the vascular anatomy can improve outcome and diminish surgical time. Several preoperative imaging techniques exist for surgical planning. Computed tomographic and magnetic resonance angiography are two emerging modalities that provide exceptional anatomic detail. Despite the growing utilization of cross-sectional imaging for preoperative planning, each modality has specific technical considerations that are necessary to consider in order to produce a quality study. PMID- 21620148 TI - Computed tomographic angiography: clinical applications. AB - There has been a move towards increasingly refined techniques for autologous breast reconstruction, and given the substantial inter-individual variability of perforator anatomy, the need for reliable, accurate methods of vascular imaging has been sought. Computed tomographic angiography (CTA) can offer a range of applications in autologous breast reconstruction to aid surgical planning and improved outcomes. This article explores the utility of CTA in imaging perforators, pedicles and recipient vessels across a wide range of flap types and donor sites. CTA has a range of clinical applications in autologous breast reconstruction, and can aid operative planning and improve outcomes. PMID- 21620149 TI - Computed tomographic angiography: assessing outcomes. AB - Perforator flaps are preferable for breast reconstruction after mastectomy in many patients. Preoperative imaging of the perforators and source vessels is desirable to reduce surgeon stress, limit donor and recipient site complications, and minimize operative time and associated costs. Computed tomographic angiography (CTA) has been shown to provide highly accurate representations of vascular anatomy with excellent spatial resolution. A critical review of the currently available literature was performed to identify the benefits of preoperative imaging (specifically CTA) in perforator flap reconstruction. PMID- 21620150 TI - Noncontrast magnetic resonance imaging for preoperative perforator mapping. AB - Identifying the position, course, and caliber of the dominant perforator is extremely valuable in the preoperative study for perforator surgery. Besides reliability, the ideal technique should offer low cost and high availability and reproducibility. It should be fast, easy to interpret, and free of morbidity. Multidetector-row computed tomography (MDTC) and magnetic resonance imaging (MRI) provide images that are easy to interpret, and assess the perforator's caliber and localization and its intramuscular course and anatomic relationships. Noncontrast MRI avoids radiation to the patient and eliminates the need for intravenous contrast medium. This article discusses this method and presents our experience. PMID- 21620151 TI - Contrast-enhanced magnetic resonance angiography. AB - With technological advances in magnetic resonance angiography (MRA), spatial resolution of 1-mm perforating vessels can reliably be visualized and accurately located in reference to patients' anatomic landmarks without exposing patients to ionizing radiation or iodinated contrast, resulting in optimal perforator selection, improved flap design, and increased surgical efficiency. As their experience with MRA in breast reconstruction has increased, the authors have made changes to their MRA protocol that allow imaging of the vasculature in multiple donor sites (buttock, abdomen, and upper thigh) in one study. This article provides details of this experience with multiple donor site contrast-enhanced MRA. PMID- 21620152 TI - Dynamic infrared thermography. AB - This article describes how dynamic infrared thermography (DIRT) can be used in autologous breast reconstruction with a deep inferior epigastric perforator flap. This noninvasive and noncontact technique for indirect monitoring of skin blood perfusion can be used in the preoperative planning and intraoperative evaluation of flap perfusion, as well as the postoperative monitoring of perfusion dynamics of DIEP flaps. DIRT provides valuable information on the perfusion physiology of perforators. PMID- 21620153 TI - Fluorescent angiography. AB - Fluorescent angiography is a simple and effective real-time tool for measurement of tissue perfusion both in and out of the operating room. It has multiple uses including: (1) identifying perforating vessels during flap planning; (2) locating primary and secondary angiosomes within a prepared flap; (3) as an aid in decision making for tissue debridement and flap creation; (4) intraoperative evaluation of microanastomoses; (5) postoperative flap monitoring, and (6) documentation of perfusion. The technology is easy to use in the hands of the operating surgeon and is safe for the patient, as it requires no radiation exposure. PMID- 21620154 TI - Near-infrared spectroscopy in autologous breast reconstruction. AB - Autologous breast reconstruction is commonly performed after mastectomy and provides a natural replacement that mimics the native breast. Although current free flap success rates exceed 95%, total flap loss can be devastating for patients. As a result, new technologies have emerged to provide an objective means for flap monitoring, with the hope that vaso-occlusive events can be detected before the clinical manifestation of a microvascular complication and the no-reflow phenomenon. This article focuses on the available data for one new technology, near-infrared spectroscopy, and its current use in clinical practice. PMID- 21620155 TI - Use of the implantable Doppler in free tissue breast reconstruction. AB - In this article, the authors discuss the advances in monitoring free tissue transfers, with a focus on the implantable Doppler system. Authors address indications and techniques for implanting the Doppler system, in addition to presenting a framework to assess the reliability and potential benefits of the implantable Doppler device. PMID- 21620156 TI - Noninvasive tissue oximetry. AB - Noninvasive tissue oximetry is playing an important role in postoperative monitoring of autologous tissue breast reconstruction flaps. It is increasingly being used intraoperatively to assist in perforator selection, tissue mapping, and assessment of mastectomy skin flap viability. This article reviews the use of tissue oximetry for intraoperative decision making, flap physiology, and postoperative monitoring, and also comments on common flap complications. PMID- 21620157 TI - Maternal mental illness and the safety and stability of maltreated children. AB - OBJECTIVE: Children of mothers with mental illness are at risk for multiple untoward outcomes, including child maltreatment and foster care placement. The purpose of this analysis was to determine the association between maternal mental illness and children's long term safety and stability. METHODS: A multi-sector administrative dataset from the Department of Social Services (DSS) and Department of Mental Health (DMH) was analyzed. The sample was 4,895 low income families (mother and child dyads) first reported to child welfare in 1993 or 1994. Families were followed until March of 2009. Dates of new report and foster care placement were obtained from DSS data. ICD-09 or ICD-10 diagnostic codes were obtained from Department of Mental Health data. Schizophrenic disorders, episodic mood disorders, anxiety disorders and personality disorders were examined. RESULTS: New reports were more likely for children of mothers with mental illness, regardless of diagnosis. While overall 67% of children had a new report over the course of their childhood, rates ranged from 80 to 90% for children of mothers with mental illness and occurred within a shorter time frame than for other children. In the multivariate models, mood (HR=1.41, p<.001) and anxiety disorders (HR=1.32, p<.05) placed children at greater risk for new reports. The proportion of children with foster placements was more than double for children of mothers with mental illness than for other children. In the multivariate model, anxiety disorders were strongly associated with the risk of placement (HR=1.75, p<.001). CONCLUSIONS AND PRACTICE IMPLICATIONS: Important differences in safety and stability were found between children of mothers with and without mental illnesses, as well as some variability across diagnoses. Since these mothers had already received services our findings suggest that access is not enough. The services they are receiving or have received may be an ineffective approach to helping them parent safely. PMID- 21620158 TI - Gender differences in filicide offense characteristics--a comprehensive register based study of child murder in two European countries. AB - OBJECTIVE: This study searched for gender differences in filicidal offense characteristics and associated variables. METHODS: In this bi-national register based study all filicide perpetrators (75 mothers and 45 fathers) and their crimes in Austria and Finland 1995-2005 were examined for putative gender differences. The assessed variables were associated with the offense characteristics, the offenders' socioeconomic and criminal history, and related stressful events. RESULTS: Mothers had previously committed violent offenses less often than fathers (5% vs. 28%, p<0.001) and they were less often employed (27% vs. 49%, p<0.05). Mothers' victims were on average younger than those of fathers; median ages of the victims were 3.4 and 6.1 years, respectively (p<0.001). Fathers were more often intoxicated during the offense (11% vs. 42%, p<0.001) and also used shooting as the method of operation more often than mothers (5% vs. 27%, p<0.001). Mothers used drowning, criminal negligence, and poisoning more often than fathers. Fathers' motives were more impulsive in nature (13% vs. 41%, p<0.001). After the killing, mothers tried to get rid of the body more often than fathers (25% vs. 7%, p<0.05). CONCLUSIONS: Fathers who commit filicide may represent at least two subgroups, the one not unlike the common homicide offender; the other, the overloaded, working and suicidal father. Mothers may include several types of offenders, one of which is the neonaticide offender. More detailed descriptions and, therefore, more research are needed. PRACTICE IMPLICATIONS: Distressed parents and families need support and health care personnel, social work and other officials need to be alert to notice fatigued parents' signs of despair, especially when several stressful experiences amass. Straightforward enquiry to the situation and even practical and psychological help may be needed for enhanced protection of children. The role of employers should also be discussed in relation to the welfare of working parents. PMID- 21620159 TI - History of abuse and neglect in patients with schizophrenia who have a history of violence. AB - OBJECTIVE: To determine the prevalence of five forms of abuse/neglect during childhood and adolescence in a group of schizophrenic patients with a history of violence. METHODS: Twenty-eight patients hospitalized in a highly secured psychiatric unit were included. Abuse and neglect during patients' growth were evaluated with the childhood trauma questionnaire (CTQ). History of substance abuse (consumption of cannabis, and/or alcohol, and/or heroin, and/or cocaine during the year that preceded the hospitalization), incarceration, and death of a close parent were also collected. RESULTS: We found that 46.4% of patients experienced at least 1 form of abuse and/or neglect during childhood and 21.4% of them had experienced more than 2 forms of abuse and/or neglect. The 2 most frequent forms of neglect and abuse were physical abuse (39.3%) and emotional neglect (17.9%). History of substance abuse was found for cannabis (57.1%), alcohol (57.1%), and cocaine and/or heroin (35.7%). We found that 42.8% of patients had 1 close relative who had died during their growth and that 41.6% of these deaths were violent. CONCLUSION: It appears important to systematically search for and assess a history of abuse and neglect during growth in schizophrenic patients with a history of violence, in order to offer specific treatments for this group of patients. PMID- 21620160 TI - Investigating geographic variation in use of psychotropic medications among youth in child welfare. AB - OBJECTIVES: Public Law (P.L.) 110-351, the "Fostering Connections to Success Act," calls for state child welfare agencies to partner with Medicaid and pediatric experts to provide planning and oversight regarding the provision of health and mental health services, including medication, to children in state custody. Recent reports, media cases, and class action lawsuits suggest over-use of psychotropic medications to address the behavioral needs of children in the child welfare system. We examined geographic variability in psychotropic medication use across US child welfare agency catchment areas to determine how rates of psychotropic medication use vary in relation to child, community, child welfare, and health system-level factors. METHODS: Cross-sectional analysis of Wave 1 data for the 92 child welfare catchment areas participating in the National Survey of Child and Adolescent Well-being, a random probability sample of 2,504 children ages 2-15 years undergoing investigation for abuse and neglect. We employed multilevel regression modeling to examine the impact of catchment level variables on medication use, controlling for child-level variables. RESULTS: Fifteen percent of children reported taking psychotropic medications. Rates of medication use across catchment areas ranged widely from 0 to 40%, a 40 fold difference. On multi-level logistic regression modeling, older age (p<.001), male gender (p<.001), emotional and behavioral problems (p<.001), and insurance (p=.05) were associated with psychotropic medication use at the child-level. At the catchment-level, stressful environment within the child welfare system was negatively associated with medication use. No other catchment-level variables examined were found to explain use. CONCLUSIONS: Striking disparities in medication use exist across catchment areas in this national sample. Of the catchment variables examined, only stressful environment was related to medication use. PRACTICE IMPLICATIONS: These findings highlight significant geographic variation in medication use that most likely reflect both under-use and over-reliance on psychotropic medication. The link between child welfare environment and medication use suggests the influence of systemic, as opposed to clinical, causes of variation in medication use. This requires greater implementation of organizational processes governing quality of care for this highly vulnerable population. PMID- 21620161 TI - Children's disclosures of sexual abuse: learning from direct inquiry. AB - OBJECTIVES: Published protocols for forensic interviewing for child sexual abuse do not include specific questions about what prompted children to tell about sexual abuse or what made them wait to tell. We, therefore, aimed to: (1) add direct inquiry about the process of a child's disclosure to a forensic interview protocol; (2) determine if children will, in fact, discuss the process that led them to tell about sexual abuse; and (3) describe the factors that children identify as either having led them to tell about sexual abuse or caused them to delay a disclosure. METHODS: Forensic interviewers were asked to incorporate questions about telling into an existing forensic interview protocol. Over a 1 year period, 191 consecutive forensic interviews of child sexual abuse victims aged 3-18 years old in which children spoke about the reasons they told about abuse or waited to tell about abuse were reviewed. Interview content related to the children's reasons for telling or for waiting to tell about abuse was extracted and analyzed using a qualitative methodology in order to capture themes directly from the children's words. RESULTS: Forensic interviewers asked children about how they came to tell about sexual abuse and if children waited to tell about abuse, and the children gave specific answers to these questions. The reasons children identified for why they chose to tell were classified into three domains: (1) disclosure as a result of internal stimuli (e.g., the child had nightmares), (2) disclosure facilitated by outside influences (e.g., the child was questioned), and (3) disclosure due to direct evidence of abuse (e.g., the child's abuse was witnessed). The barriers to disclosure identified by the children were categorized into five groups: (1) threats made by the perpetrator (e.g., the child was told (s)he would get in trouble if (s)he told), (2) fears (e.g., the child was afraid something bad would happen if (s)he told), (3) lack of opportunity (e.g., the child felt the opportunity to disclose never presented), (4) lack of understanding (e.g., the child failed to recognize abusive behavior as unacceptable), and (5) relationship with the perpetrator (e.g., the child thought the perpetrator was a friend). CONCLUSIONS: Specific reasons that individual children identify for why they told and why they waited to tell about sexual abuse can be obtained by direct inquiry during forensic interviews for suspected child sexual abuse. PRACTICE IMPLICATIONS: When asked, children identified the first person they told and offered varied and specific reasons for why they told and why they waited to tell about sexual abuse. Understanding why children disclose their abuse and why they wait to disclose will assist both professionals and families. Investigators and those who care for sexually abused children will gain insight into the specific barrier that the sexually abused child overcame to disclose. Prosecutors will be able to use this information to explain to juries why the child may have delayed his or her disclosure. Parents who struggle to understand why their child disclosed to someone else or waited to disclose will have a better understanding of their child's decisions. PMID- 21620162 TI - Self-defining as sexually abused and adult sexual risk behavior: results from a cross-sectional survey of women attending an STD clinic. AB - OBJECTIVE: Childhood sexual abuse (CSA) is associated with increased sexual risk behavior in adulthood, and this association may be mediated by traumagenic dynamics constructs (i.e., traumatic sexualization, trust, guilt, and powerlessness). However, few studies have investigated whether such relationships hold for women who do not identify as having experienced CSA despite meeting objective criteria that CSA occurred. This study sought to determine whether individuals who met research criteria for CSA and who self-defined as sexually abused differed on traumagenic dynamics constructs and current sexual risk behavior from individuals who met research criteria for CSA and who did not self define as sexually abused. METHODS: Participants were 481 women recruited from a publicly funded STD clinic. Participants completed a computerized survey assessing childhood sexual experiences and adult sexual risk behavior. RESULTS: Of the total sample, 206 (43%) met research criteria for CSA. Of the women meeting research criteria for CSA, 142 (69%) self-defined as sexually abused. Women who met research criteria for CSA reported more traumatic sexualization, more trust of a partner, more powerlessness, less sexual guilt, more episodes of unprotected sex, more sex partners, and greater likelihood of sex trading, compared to women who did not meet research criteria for CSA. Among women meeting research criteria, those who self-defined as sexually abused did not differ from those who did not self-define on any of the traumagenic dynamics constructs or on current sexual risk behavior, controlling for CSA characteristics. CONCLUSIONS: Individuals who were sexually abused as children by behavioral research criteria are at risk for engaging in sexual risk behavior as adults, regardless of whether or not they perceive the experience to be CSA. Future research is needed to understand how non-definers perceive these childhood sexual experiences. PMID- 21620163 TI - Intergenerational transmission of sexual victimization vulnerability as mediated via parenting. AB - OBJECTIVES: Previous research suggests that women's early sexual victimization experiences may influence their parenting behaviors and increase the vulnerability of their children to being sexually victimized. The current study considered whether mother's sexual victimization experiences, in childhood and after age 14, were associated with the sexual victimization experiences reported by their adolescent daughters, and if so, whether these effects were mediated via parenting behaviors. METHODS: The proposed model was examined using a community sample of 913 mothers and their college-bound daughters, recruited by telephone at the time of the daughter's high school graduation. Daughters reported on their experiences of adolescent sexual victimization and perceptions of mothers' parenting in four domains: connectedness, communication effectiveness, monitoring, and approval of sex. Mothers provided self-reports of their lifetime experiences of sexual victimization. RESULTS: Consistent with hypotheses, mothers' victimization was positively associated with their daughters' victimization. The effect of mothers' childhood sexual abuse was direct, whereas the effect of mothers' victimization after age 14 was mediated via daughters' perceptions of mothers' monitoring and greater approval of adolescent sexual activity. Comparison of the prevalence of specific victimization experiences indicated that mothers were more likely to report forcible rape over their lifetimes; however, daughters were more likely to report unwanted contact and incapacitated rape. CONCLUSIONS: Findings suggest that even in a highly functional community sample, mothers' sexual victimization experiences are significantly associated with aspects of their parenting behavior and with their daughters' own experiences of adolescent sexual victimization. PMID- 21620164 TI - Intrafamilial conflict and emotional well-being: a population based study among Icelandic adolescents. AB - OBJECTIVES: During intrafamilial conflicts children are often innocent bystanders, caught in the crossfire. In such situations, they are at increased risk to become directly involved in abusive verbal behavior of the perpetrator, and exposed to being shouted or yelled at, threatened, rejected and even physically abused. The present study has two main objectives: (1) ascertain a national base rate of intrafamilial conflicts and physical violence at home among Icelandic adolescents; and (2) to investigate the association of witnessing and/or having been a part of intrafamilial conflict or physical violence at home with variables that relate to mental health and well-being. METHODS: The participants were 3,515 students, 14- and 15-year-old, in the national compulsory school system in Iceland. As a part of the 2003 ESPAD survey, each pupil was asked about experiences of severe verbal arguments and physical violence at home as well as their background, behaviors, and mental health assessed with the use of tested measurement scales such as the Symptom Distress Checklist 90 (SCL-90) and the Rosenberg Self-Esteem Scale. RESULTS: About 22% of the participants stated that they had witnessed a severe verbal argument between parents and 34% stated that they had been involved in a severe verbal argument with parents. This rate was slightly higher for girls compared to boys. All together 7% of adolescents had witnessed physical violence at home where an adult was involved and 6% of the participants stated that they had experiences of being involved in physical violence at home where an adult was involved. Witnessing or being involved in severe verbal arguments at home and/or witnessing or being involved in physical violence with an adult was significantly associated with greater levels of depression, anger, and anxiety, and negatively related with self-esteem (p<0.01). CONCLUSIONS: Many adolescents in Iceland witness severe parental verbal arguments or physical violence between adults in their homes and some are directly involved in such acts. It affects their long-term emotional and behavioral development and well-being. PRACTICE IMPLICATIONS: Preventive measures have to be implemented at an early age and should include, but not be limited to, information on disciplining and upbringing of children and the negative impact of intrafamilial conflicts on the long-term health of their children. Due attention should be given to the health and well-being of children where such violence is known to occur. PMID- 21620165 TI - Student victimization by teachers in Taiwan: prevalence and associations. AB - OBJECTIVES: This paper reports on the prevalence of student victimization by teachers in junior high schools in a Chinese cultural context (Taiwan) and examines how student demographic variables (gender, grade level, and family socioeconomic status) and school social experiences (student-teacher relationships and involvement with at-risk peers) are associated with such victimization. METHODS: Data were obtained from a large-scale random sample of 1,376 junior-high students (grades 7-9) in the city of Taichung, Taiwan. Students were given an anonymous structured questionnaire, including items regarding basic demographics and school social experiences. RESULTS: Overall, 26.9% of students reported having been maltreated by teachers at least 1 time in the previous semester. Hitting, beating, or slapping was the most common maltreatment, and the most vulnerable students were boys and senior students. Students who perceived that student-teacher relationships were poor, and those who were involved with at risk peers, were more likely to report victimization. CONCLUSION: Although there are clear guidelines and regulations prohibiting teacher aggression against students, Taiwanese students are still exposed to high levels of maltreatment. The findings provide empirical evidence to support school social workers and policymakers in taking immediate action to educate politicians, the general public, and the media about the severity of student victimization by teachers as well as to build up mechanisms to supervise the government's enforcement of regulation. These findings clearly imply that promoting positive social experiences for students is crucial for successful intervention. PMID- 21620284 TI - Blocking the renin-angiotensin system--history and controversies. PMID- 21620285 TI - The evolution of angiotensin blockade in the management of cardiovascular disease. AB - Dysregulation of the renin-angiotensin system is implicated in many cardiovascular and renal pathologies. Discovery of the renin-angiotensin system represents a major advance in the understanding of hypertension and cardiovascular disease, leading to the development of the anti-angiotensin medications: angiotensin-converting enzyme inhibitors, angiotensin receptor blockers and direct renin inhibitors. Clinical trials have shown that drugs in each of these classes have a protective effect on vascular organs that surpass the protection associated with the lowering of blood pressure alone. PMID- 21620286 TI - Reviewing the benefits of angiotensin-converting enzyme inhibitors and angiotensin receptor blockers in diabetic nephropathy--are they drug specific or class specific? AB - Hypertension is prevalent in patients with type 2 diabetes mellitus and contributes to substantial morbidity and mortality. Preventing microalbuminuria (MAU) is important in these patients because MAU is associated with an increased risk of end-stage renal disease and cardiovascular events. Current hypertension treatment recommendations are based on drug 'class' effects. For the treatment of hypertension in patients with diabetes, important agent-specific effects should be brought into consideration in the choice of drug. For these patients, and especially in the presence of kidney disease, the choice of agent must effectively target not only blood pressure reduction, but also prevention and treatment of MAU for optimal organ outcomes. A review of the current evidence suggests a diversity of MAU-associated risk reduction according to agent within the angiotensin receptor blocker and angiotensin-converting enzyme inhibitor classes, ranging from nonefficacy to significant efficacy. Recommendations should clearly state absolute risk reduction according to agent, based on the best available evidence. PMID- 21620287 TI - Reviewing the future of renin-angiotensin system blockade: the role of angiotensin-converting enzyme inhibitors and angiotensin receptor blockers in the prevention of atrial fibrillation. AB - Atrial fibrillation (AF) is the most common sustained arrhythmia encountered in clinical practice, affecting approximately 250,000 Canadians and accounting for more than 120,000 hospitalizations each year. Atrial remodelling, which is at least partially induced by activation of the renin-angiotensin system (RAS), can be reversed by RAS blockade using angiotensin-converting enzyme inhibitors or angiotensin receptor blockers. While RAS antagonists are not considered to be conventional anti-arrhythmic agents, several studies have shown that they may reduce the incidence of AF in patients with heart failure, after myocardial infarction with reduced ejection fraction, and in hypertensive patients. Angiotensin-converting enzyme inhibitors and angiotensin receptor blockers have also been used in association with amiodarone to facilitate sinus rhythm maintenance after electrical cardioversion. Whether RAS blockade prevents recurrent AF in patients at high cardiovascular risk remains unknown. PMID- 21620288 TI - Validating the patient safety indicators in the Veterans Health Administration: are they ready for prime time? PMID- 21620289 TI - Variation in quality of care after emergency general surgery procedures in the elderly. AB - BACKGROUND: The elderly (age >=65 years) comprise an increasing proportion of patients undergoing emergency general surgery (EGS) procedures and have distinct needs compared with the young. We postulated that the needs of the elderly require different processes of care than those required for the young to assure optimal outcomes. To explore this hypothesis, we evaluated 30-day outcomes following EGS procedures in the young and the elderly and determined whether hospital performance was consistent across these 2 age strata. STUDY DESIGN: With data from the American College of Surgeons National Surgical Quality Improvement Program (2005 to 2008), regression models were constructed for serious morbidity and mortality for all patients undergoing EGS procedures and separately for young and elderly patients. These models allowed for estimation of the risk of adverse outcomes associated with advanced age and the generation of hospital-level observed to expected (O/E) ratios. We evaluated the correlation between hospital O/E ratios for the young and the elderly and the concordance of outlier status (hospitals with CIs of O/E ratios excluding 1) with weighted kappa across these 2 age groups. RESULTS: Among 68,003 procedures at 186 hospitals, elderly patients had a higher crude and adjusted risk for serious morbidity (27.9% versus 9.7%, p < 0.0001; odds ratio 1.17, 95% CI 1.10 to 1.24) and mortality (15.2% versus 2.5%, p < 0.0001; odds ratio 2.29, 95% CI 2.09 to 2.51). When outcomes for elderly versus younger patients were compared, there was fair to moderate agreement on hospital performance for serious morbidity (r = 0.43; kappa = 0.30) but not for mortality (r = 0.10; kappa = 0.17). CONCLUSIONS: Elderly patients are at substantially greater risk for adverse events following EGS procedures. Hospitals had only slight agreement in mortality outcomes in the elderly compared with those in young patients. Processes of care that may account for this disparity should be further investigated. PMID- 21620290 TI - Effect of preoperative smoking cessation interventions on postoperative complications. PMID- 21620292 TI - Feasibility and safety of a new robotic thyroidectomy through a gasless, transaxillary single-incision approach. PMID- 21620293 TI - Limiting the surgical indications for liver hemangiomas may help surgeons and patients. PMID- 21620294 TI - Online education in health care: quality and convenience. PMID- 21620296 TI - [The role of chemotherapy in the management of bladder cancer]. AB - Bladder cancer represents for man the second genitourinary cancer after prostate cancer. Urothelial carcinoma is the most predominant histological type. In up to 70% of the cases, the diagnosis of bladder cancer is performed at early stages (Ta-T1). In this situation, the treatment of the disease is the transurethral resection with or without intravesical treatment (BCG, Amiticyne). In advanced disease, treatment is essentially palliative with chemotherapy based on cisplatin type MVAC (methotrexate, vinblastine, doxorubicin, and cisplatin) or gemcitabine cisplatin. In invasive stages (T2-T3-T4), the radical cystoprostatectomy combined with urinary diversion for man, and the pelvectomy for woman are the gold standard. However, over 50% of these patients experienced metastatic recurrence during their evolution, which prompted investigators last 10 years to assess the value of neoadjuvant chemotherapy in their management. Indeed, neoadjuvant chemotherapy is now recognized as a standard by numerous American and European institutions. However, adjuvant chemotherapy remains controversial even for patients with lymph node involvement. The purpose of this literature review is to highlight the role of chemotherapy in the management of urothelial carcinoma of bladder with locally advanced and metastatic disease. The role of targeted therapies alone, in combination with chemotherapy, and in maintenance, is being evaluated. PMID- 21620297 TI - [Usefulness of real time transrectal ultrasonography to monitor the Greenlight XPS((r)) (180W) laser prostatectomy]. AB - We report a technical improvement of the Greenlight((r)) XPS prostatectomy by using real time transrectal ultrasonography (TRUS) during the procedure. The TRUS probe lays on a flexible support fastened to the operative table. A wide screen with double display allows the surgeon to control the progress of the procedure with simultaneous ultrasonographic and endoscopic views. The depth of vaporization clearly appears as an hyperechogenic line together with an enlargement of the vaporization area. Real time TRUS monitoring during photovaporization of BPH decreases the learning curve, the risk of prostatic capsule perforation and bleeding, and allows to check the completeness of the procedure. PMID- 21620298 TI - [When do we have to stop a prostatic photovaporization with the greenlight laser?]. PMID- 21620299 TI - [Ureteral stenosis after renal transplantation: Risk factors and impact on survival]. AB - OBJECTIVES: To identify the risk factors for ureteral stenosis after renal transplantation and to evaluate their impact on both graft and patient survival. PATIENTS AND METHODS: This retrospective study included 789 kidney transplants among 782 patients performed at our institution between 1995 and 2007. The parameters studied included the characteristics of the donor, recipient and transplant, the surgical variables, the elements of the monitoring process and a graft and patient survival. RESULTS: The ureteral stenosis rate after renal transplantation was found to be 6.5%, and the ureterovesical junction was the most common location (68%). A univariate analysis showed that this complication was significantly associated with a higher donor age (P=0.01), abnormal graft revascularisation (P=0.032) and DGF (Delay Graft Function) (P=0.05). In multivariate analysis, only donor age (P=0.001) and abnormal graft revascularisation (P=0.035) were independent risk factors for ureteral stenosis after renal transplantation. When ureteral stenosis was treated, an analysis of the survival curves according to the Kaplan-Meier method did not reveal significant differences either in graft survival (P=0.518) or overall survival of the patients (P=0.614) as compared to the control group. CONCLUSIONS: In the present study, donor age and abnormal graft revascularisation were independent risk factors for ureteral stenosis after renal transplantation. This result is a strong argument for an ischemic component in the genesis of ureteral stenosis after renal transplantation, which should help to identify patients at risk. PMID- 21620300 TI - [The thermoformable spiral metallic stents in the treatment of localized ureteral stenosis: an alternative to JJ stent? Prospective multicenter study]. AB - INTRODUCTION: Evaluation of the effectiveness and tolerance of thermoformable metallic spiral stents Memokath((r)) 051 (Bard, Pnn Medical) in the treatment of localized ureteral stenosis in non-operable patients who have JJ ureteral stents. MATERIAL AND METHOD: Prospective, descriptive and multicenter study of patients with ureteral strictures treated with metallic ureteral stents Memokath((r)) 051. Assessment criteria (recurrent stenotic, permeability, tolerance) were measured by clinical, biological and radiological examination at 1 month, and then every 3 months. RESULTS: Fifteen stents (average length: 9.15 cm, range 6-15 cm) were implanted in 14 patients (mean age: 55 years, range: 38-72 years) with secondary suspended ureteral stenosis during 2 years in two centers. The median follow-up was 11 months (range 6 to 24 months). Technical difficulty was observed with two patients. Stents are still up in four patients. The stenosis recurred in four patients with spontaneous progression of stenosis but without endoprosthetics tissue invasion. Two and three migration were observed with spontaneous expulsions. Two lower urinary infections and one high occurred, resolved on antibiotic therapy, no inlay or hematuria, no pain (mean VAS score=3/10) or urinary disorders of the lower unit have been identified. CONCLUSION: Stents Memokath((r)) 051 are well tolered and seem to position themselves as an interesting alternative to JJ ureteral stent in some frails patients. The refinement of contraindication should help to improve the stent's efficacity and to reduce the risk of migration and expulsion. PMID- 21620301 TI - [Influence of occupational exposures in nonmuscle infiltration bladder cancer: preliminary results of a prospective study from October 2005 to February 2009]. AB - Bladder cancer is a common condition in industrialized countries. If tobacco is still the main risk factor in lung cancer, occupational exposures carcinogens should not be underestimated. GOAL: The significant excess of bladder cancer in the north part of France, with high manufacture concentration likely to have employees exposed to bladder carcinogens, has led us to study the influence of these exposures in the natural history of bladder cancer. PATIENTS AND METHODS: We prospectively conducted a descriptive case-control study. A questionnaire was developed by the department of occupational disease and clinical, radiological, histological, therapeutic data were registered at the University Hospital of Lille. From October 2005 to February 2009, 69 patients were included in the study, 37 exposed to occupational carcinogens and 32 in the control group. RESULTS: Mean age was 61.6 years vs. 61.8 years and the sex ratio of 7.33 men to one woman vs. one woman for three men respectively in the two groups. The average age of patients exposed to polycyclic aromatic hydrocarbons was 59.7 years. Smokers were 86.5% and 87.5% respectively. Follow-up was 38.4 and 32.9 months respectively. Nonmuscle invasive bladder cancer were more frequent (P=0.019) in the exposed group (84.4%) than in the unexposed group (67.8%) even after adjustment for smoking (P=0.0142). The histological type, grade, presence of CIS, the early recurrence at 3 months, the number of lesions at diagnosis does not differ in the two groups even after adjustment for smoking or after subgroup analysis of the most frequent exposure. The exposure to polycyclic aromatic hydrocarbons (62%) and aromatic amines (37.8%) were the most represented. Of 37 patients, 13 (35%) were making a statement as an occupational disease (eight according to Table 15 ter, two according to Table 16 bis and two presented to IRB). To date one single patient is recognized as an occupational disease. CONCLUSION: We did not observe any worsening of the prognosis of bladder cancer following occupational carcinogen exposure except for the mean age at diagnosis. The small size of the population studied and the importance of smoking partner have hampered the analysis of occupational exposures. PMID- 21620302 TI - [Variation of urinary PCA3 following transrectal ultrasound-guided prostate biopsy]. AB - INTRODUCTION: Serum PSA is known to rise slightly following an attentive digital rectal examination (DRE) and dramatically following prostatic biopsy. The aim of this study was to evaluate the PCA3 response in these situations. PATIENTS AND METHODS: In 15 consecutive men undergoing transrectal ultrasound-guided needle biopsy of the prostate and who gave their informed consent, urinary PCA3 was determined twice: at a first consultation, urine being sampled immediately after an attentive DRE and second within 2 hours after the biopsy. The mean interval between the two samplings was 14 days (median 15). PCA3 measurements were centralized and performed by the same biologist. At least twelve cores were taken using a biopsy gun with an 18-gauge needle. Changes in PCA3 levels were studied. RESULTS: Mean age of the 15 men was 67.3 years (range 50.9-79.1). Mean (median) pre-biopsy total and %free PSA were respectively 6.6 ng/ml (5.7) and 15.8% (15.5). Mean prostate volume was 43.6 cm(3). Seven patients complained of mild LUTS. DRE was suspicious in eight patients. Of the 15 men, 6 (40%) had adenocarcinoma on biopsy (all clinically confined to the prostate). Median (range) Gleason score was 6 (6-7). Median PCA3 score (range) before and after prostatic biopsy were respectively 36 (9-287) and 27 (5-287) with no significant difference between the two groups (sign test for matched series p > 0.05). The median variation between pre- and post-biopsy PCA3 was -18%. When considering a PCA3 cut-off of 35, two patients changed group: one patient had 51 before and 31 after (PSA 4.6; no cancer on prostate biopsy) and the second had 36 before and 27 after (PSA 5.6; low-risk PCa). The figure represents the PCA3 values for each case (squares for the pre-biopsy and diamonds for the post-biopsy). When considering only the six patients with PCA, median (mean) PCA3 score before and after prostatic biopsy were respectively 51.5 (60.8) and 44.5 (54.8) with no significant difference between the two groups (sign test for matched series p > 0.5) and a median variation between pre- and post-biopsy PCA3 of 1.5%. CONCLUSIONS: Prostate biopsy did not alter significantly urinary PCA3 value. This confirms what was theoretically expected. PMID- 21620303 TI - [Treatment of benign prostatic hyperplasia by Prostiva((r)): about a series of 76 cases]. AB - PURPOSE: To evaluate the efficacy and morbidity of BPH treatment by radiofrequency in the general population and attempt to identify factors of prognostic value for this technique. METHODS: This retrospective study examined 76 consecutive patients of mean age 68.9 years with symptomatic BPH in failure of medical treatment and treated at University Hospital of Caen between April 2004 and October 2008. Urine status was assessed before and after treatment with radiofrequency by the international prostate symptom score (IPSS), the quality of life (QOL) score related to urinary symptoms, the measurement of maximum urinary flow (Qmax) and residual postvoid volume (RPV). Erectile function was assessed by the International Index of Erectile Function (IIEF-5). The primary endpoint was treatment failure by radiofrequency regarded as the reintroduction of medical treatment or the need for surgical treatment. The secondary endpoints were the improvement of IPSS and QOL score, Qmax and RPV. Morbidity per and post-operative was also evaluated. RESULTS: A significant improvement in voiding status was observed with an average IPSS score from 24.7 to 15.1 (P<0.0001), and a QOL score from 5.2 to 2.9 (P<0,0001) in all patients. The Qmax and the RPV were significantly improved from 8.6 to 13.1ml/s (P<0.0001) and 177 to 100ml (P=0.0002) respectively. The IIEF-5 score was improved from 9.04 to 9.97 (P=0.0164). Twenty-five percent of acute urinary retention, 7.9% of haematuria and 1.3% of prostatitis were observed postoperatively. The overall retreatment rate was 46% after 14.6 months mean follow-up. When stratified into groups, patients with the inclusion criteria to this treatment showed a treatment rates of 34.8% after 16.2 months mean follow-up against 51% after 13.8 months of monitoring patients outside the inclusion criteria. Without showing significant difference between retentionist and no retentionist concerning subjective and objective parameters, the rate of retreat was more important for retentionist (51.2% after 11.8 months of mean follow against 39.4% after 18.4 months) with a significant difference on the actuarial cumulative survival curve without reprocessing. There was no significant difference found in patients with a median lobe. The results showed an improvement in Qmax significantly greater in the group with prostates less than 60 g without being able to demonstrate significant differences in other parameters. The retreatment rates in this group was 44.4% after 14.7 months of follow-up against 53.8% after 13.8 months of follow-up in the group of patients with prostate over 60 g. Moreover, the irritative component made a better initial response to treatment with a need for retreatment delayed compared to the group of patients with obstructive symptom, without noting significant differences in terms of improvement of objective and subjective parameters. The rate of retreatment was 63.6% after 16.4 months of mean follow and 51.6% after 11.2 months in the "obstructive" group. CONCLUSION: High rates of reprocessing are observed in the treatment of BPH by radiofrequency if inclusion criteria are not respected. PMID- 21620304 TI - [Laparoscopic sacrocolpopexy results contingent on mesh position]. AB - OBJECTIVES: Do the number and the position of meshes in laparoscopic sacrocolpopexy influence anatomical or functional postoperative results in genital prolapse treatment? PATIENTS AND METHODS: Ninety patients were treated for genital prolapse by laparoscopic sacrocolpopexy between January 1998 and 2007. Eleven had an anterior single mesh, 36 a single posterior mesh and 43 a double mesh. RESULTS: Four patients with late postoperative complications needed a new surgical procedure. Three of them had a double mesh. Thirteen anatomical recurrences (14 %) were found. Eleven recurrences had a single posterior mesh and eight needed a new surgical procedure. Two other recurrences had a double mesh. Only one needed a new surgical procedure. No recurrence was noticed in the anterior single mesh group. The observed pelvic floor dysfunction rates were respectively for the single anterior mesh group, posterior single mesh group and double mesh group: constipation 20 % 64 % 35 %, anal incontinence 0 % 14 %/2 %, urgency 0 %/8 %/12 %, stress urinary incontinence 27 % 14 %/31 %. CONCLUSION: Double mesh reduced anatomical recurrence, but increased surgical complications and postoperative dysfunctions. PMID- 21620305 TI - [Renal-pleural fistula after radiofrequency ablation of renal tumor in VHL patient]. AB - Radiofrequency is a minimally invasive therapy allowing tumor destruction by applying physical means to the core of the lesion. There is a particular indication for the hereditary already surgically treated renal carcinomas like Von Hippel-Lindau's disease. We present a case of renal-pleural fistula developed after a percutaneous radiofrequency ablation under computed tomography (CT) guidance of a renal tumor in a VHL female patient with a renal cell carcinoma of the upper pole of the left kidney. The kidney manifestations begin at 20-year-old with the appearance of cystic lesion at the lower pole of the left kidney. At 30 year-old, a computed tomography study revealed a solid lesion arising from a cyst. The patient underwent a partial nephrectomy by flank incision. Follow-up studies discovered three solid lesions of the upper pole of the left kidney. The patient undertook a radiofrequency ablation of these lesions. Follow-up control showed a contrast enhancement of one of the three lesions treated. Under this condition another course of RF was performed, complicated by a renal-pleural fistula. A conservative management of this iatrogenic fistula was attempted combining a water restriction and the insertion of a ureteral catheter. Three weeks were necessary until the fistula completely regress. PMID- 21620306 TI - [Hyperattenuating renal mass]. AB - Leiomyoma is a frequently uterine tumour. Its location on the urinary tract is rare, making its iconographic diagnosis difficult. The ablation is often realized, allowing the histological confirmation. Renal leiomyoma have good prognosis. In our patient, the CT scan discovered a spontaneously hyperattenuating renal mass, raising after injection of contrast, at a 48-year old patient. MRI revealed in particular a hyposignal T2. Because of its capsular location, the possibility of a renal leiomyoma had been envisaged without being able to eliminate a malignant lesion. The histological exam confirms this benign hypothesis. So leiomyoma is a rarely renal tumour, of excellent prognosis. The progress of the imaging allows the characterization of these hurts today and could, can be, in a near future, be an alternative at surgery. PMID- 21620307 TI - ASPMN continues to advance the science of pain management. PMID- 21620308 TI - Concept analysis of nurses' identification of pain in demented patients in a nursing home: development of a hybrid model. AB - Pain is a subjective feeling, with no known biologic markers. Proof of its presence and measurement of intensity rely entirely on self-reporting by the patient. The hampered or abrogated ability of demented patients to report their pain is a major difficulty in pain assessment and management. The purpose of this study was to clarify and conceptualize pain identification in demented patients by nurses. The hybrid model of concept development was used in the development of a conceptual structure of pain in demented patients. Data were collected by literature review (theoretical phase) and among nurses caring for demented patients in three nursing homes in South Korea (fieldwork phase). The 13 nurses involved each reported >3 years' nursing home experience. In a hybrid model, pain identification in demented patients by nurses constituted an active daily process of integrating patient expressional cues during periods of pain and pain relief and involving three dimensions: identification schemes based on the stage and type of dementia, connecting assessments after each intervention, and cognitive efforts to establish the origin of pain. Identification of pain in demented patients by nurses is a complex process. More research is needed to formulate an assessment tool and pain management strategies for patients with dementia. PMID- 21620309 TI - Nondrug therapies for pain management among rural older adults. AB - This quasiexperimental two-group pilot study tested an intervention aimed at educating older adults in rural communities about the appropriate use of nondrug treatments for pain. Earlier data reveal that older adults use significantly less nonpharmacologic modalities than their younger counterparts, and that pain self treatment is prevalent in rural areas. Individuals aged >=60 years who experienced pain in the preceding 2 weeks were recruited from rural Midwestern communities through the use of flyers and information sessions at hospitals, churches, and community organizations. Upon enrollment, participants selected a date for an educational session, which was randomized to the experimental or control condition. All participants (n = 53) completed a series of questionnaires (Brief Pain Inventory, Symptom Distress Scale, Perceived Control Scale) at the initial educational session (T1) and at a two-week follow-up session (T2). Participants in the control and experimental groups attended a 30-minute educational session on safe use of over-the-counter medications; the experimental group also received an additional 30-minute session on safe and effective use of heat, cold, and relaxation breathing. Hot and cold packs and relaxation breathing instruction were provided for use over the 2-week period. There was a significant increase in the use of all nondrug treatments and a decrease in pain-related distress and current pain scores in the experimental group compared with the control group. This study informs nurses and other health care providers on the value of education for use of nondrug therapies in conjunction with pharmacologic pain management among rural older adults. PMID- 21620311 TI - Procedural pain management: a position statement with clinical practice recommendations. AB - The American Society for Pain Management Nursing (ASPMN) has developed a position statement and clinical practice recommendations related to procedural preparation and comfort management. Procedures potentially produce pain and anxiety, both of which should be assessed and addressed before the procedure begins. This position statement refers to "comfort management" as incorporating the management of pain, anxiety, and any other discomforts that may occur with procedures. It is the position of ASPMN that nurses and other health care professionals advocate and intervene based on the needs of the patient, setting, and situation, to provide optimal comfort management before, during, and after procedures. Furthermore, ASPMN does not condone procedures being performed without the implementation of planned comfort assessment and management. In addition to outlining this position with supporting evidence, this paper reviews the ethical considerations regarding procedural comfort management and provides recommendations for nonpharmacologic and pharmacologic management during all phases of the procedure. An appendix provides a summary of this position statement and clinical practice recommendations. PMID- 21620310 TI - Treatment of pain in children after limb-sparing surgery: an institution's 26 year experience. AB - A significant proportion of patients report long-term pain that is >=5 on a 0-10 intensity scale after limb-sparing surgery for malignancies of the long bones. Patients experience several distinct types of pain after limb-sparing surgery which constitute a complex clinical entity. This retrospective study examined 26 years of experience in a pediatric institution (1981-2007) in pain management as long as 6 months after limb-sparing surgery and reviewed the historical evolution of pain interventions. One hundred fifty patients underwent 151 limb-salvage surgeries for bone cancer of the extremities in this series. Pain treatment increased progressively in complexity. Therapies included opioids, nonsteroidal antiinflammatory drugs, acetaminophen-opioid combinations, postoperative continuous epidural infusion, anticonvulsants and tricyclic antidepressants for neuropathic pain, local anesthetic wound catheters, and continuous peripheral nerve block catheters. Management of pain after limb-sparing surgery has evolved over the 26 years of this review. It currently relies on multiple "layers" of pharmacologic and nonpharmacologic strategies to address the complex mixed nociceptive and neuropathic mechanisms of pain in this patient population. PMID- 21620312 TI - Severe pain related to Klippel-Trenaunay-Weber Syndrome: a case study. AB - Klippel-Trenaunay-Weber Syndrome (KTWS) is characterized by a combination of vascular malformations of the skin, abnormalities of the venous and lymphatic systems, and limb enlargement due to bone and soft tissue hypertrophy. The precise etiology is unknown. There is typically no evidence of arteriovenous shunting. Venous malformations are present at birth, are known to progress as the patient grows, and do not regress. Blood flow through these malformations is sluggish, causing ischemia and thrombotic painful events. Nerve blocks are often used to treat pain that is unrelieved by other pharmacologic and nonpharmacologic measures. In this particular case, a stellate ganglion nerve block was the treatment of choice. The stellate ganglion is part of the sympathetic nervous system made up of the inferior cervical and first thoracic ganglia. The nerve block was performed under fluoroscopy by an anesthesiologist board certified in pediatric pain management. The goal of the block was to relieve the patient's pain by calming painful nerve impulses produced by the stellate ganglion and improving blood flow to the area by vasodilating the vessels distal to the site of the block. The goal of this report was to impress upon health care professionals the effectiveness of nerve blocks in improving and sometimes eliminating a patient's pain, as well as the importance of exhausting all options to alleviate/improve pain in our patients. To protect the privacy of this patient and family, the details of the actual case were revised. PMID- 21620313 TI - Policy and practice developments in the implementation of shared decision making: an international perspective. PMID- 21620314 TI - Shared decision making in Australia in 2011. AB - The paper describes the current position of shared decision making (SDM) within the Australian healthcare system. SDM POLICY IN AUSTRALIA: Support for SDM exists through guidelines and policy documents, and is strongly endorsed by consumer organisations; however, there is no clear overarching policy framework for SDM in Australia. TOOLS FOR SDM: There are limited tools available for SDM in clinical practice. Access to tools exists through some Australian health research and consumer organisation websites but the use of tools remains idiosyncratic. IMPLEMENTATION OF SDM: Comparatively little has been achieved in the implementation of SDM in Australia. Although there is wide recognition that consumer involvement in health decisions is important, provision of resources and infrastructure to achieve it is limited, and there is no clear strategy to support implementation within the healthcare system. SDM IN THE FUTURE: Current reforms to the healthcare system may enable a more centralised approach to implementation of SDM in the future. A new federally funded consumer health information organisation may assist by providing a central point through which SDM interventions may be made available to the Australian public and the Australian Charter of Rights has the potential to provide a national framework for consumer involvement. However, priority needs to be given to SDM by both federal and state governments with greater investment in SDM research and in activities to support implementation in clinical practice. PMID- 21620315 TI - Shared decision making in Brazil: history and current discussion. AB - There is no SDM in clinical practice in Brazil. The first steps have been taken towards research and tool development recently. Likewise, our society is starting to get involved with decision making in health care. This paper aims to offer an overview of the Brazilian health system history, its values, and its influence on SDM. The participative social control concept is introduced as a result of the movement against the dictatorship era. In addition, the influence of social changes on the Medical Ethical Code is delineated. SDM state of the art in Brazil is also discussed and the challenges to implement it on clinical practice are described. Regardless the challenges, it is possible to make a positive assessment of SDM in Brazil. PMID- 21620316 TI - Moving SDM forward in Canada: milestones, public involvement, and barriers that remain. AB - Canada's approach to shared decision making (SDM) is as disparate as its healthcare system; a conglomerate of 14 public plans at various administrative levels. SDM initiatives are taking place in different pockets of the country and are in different stages of development. The most advanced provincial initiative is occurring in Saskatchewan, where in 2010 the provincial government prepare to introduce patient decision aids into certain surgical specialties. With regard to decision support tools for patients, perhaps the most active entity is the Patient Decision Aids Research Group in Ottawa, Ontario. This group maintains a public inventory of decision aids ranked according to International Patient Decision Aid Standards and has developed the generic Ottawa Personal Decision Guide, as well as a toolkit for integrating decision aids in clinical practice. All of these tools are publicly available free of charge. Professional interest in SDM in Canada is not yet widespread, but Canada's principal health research funding agency is sponsoring several important SDM projects. Researchers from institutions across the country are promoting SDM through continuing professional development programs and other interventions in fields as varied as primary care, dietary medicine and workplace rehabilitation. Still, the future of SDM in Canada remains uncertain. Canada's provincially based structure obliges promoters to work with each province separately, and the recent growth of private healthcare risks dissipating efforts to implement SDM. PMID- 21620317 TI - Shared decision making in Chile: supportive policies and research initiatives. AB - WHAT ABOUT POLICY REGARDING SDM? Since 1999, there has been a small but growing interest by academics, the government, and society as a whole in strengthening patients' and professionals' involvement in shared decision making (SDM). Two governmental policy documents that indicate support for SDM are (1) Health Reform in 2003 and (2) Sanitary Objectives 2011-2020, which includes a brief section on client participation and SDM. WHAT ABOUT TOOLS - DECISION SUPPORT FOR PATIENTS? Research by Chilean academics has highlighted the patients' desire to participate in health decisions and effective approaches for enhancing health professionals' skills in interprofessional SDM; however, little has been done to support this need and the work is centralised in only one academic institution. Decision support tools and coaching interventions are limited to patients considering decisions about managing type 2 diabetes. WHAT ABOUT PROFESSIONAL INTEREST AND IMPLEMENTATION? Although there is increasing attention to studying patients' participation and involvement on their healthcare, little has been studied in relation to professionals' interest in SDM. As well, there are significant challenges for implementation of a country-wide SDM policy. WHAT DOES THE FUTURE LOOK LIKE? The future looks promising given the new health policies, local Chilean research projects, and international initiatives. Collaboration between health professionals, academics, and government policy makers, with public involvement needs to be strengthened in order to promote concrete strategies to implement SDM in Chile. PMID- 21620318 TI - Shared decision making in the physician-patient encounter in France: a general overview in 2011. AB - WHAT ABOUT POLICY REGARDING SDM? There is a social demand in France for more healthcare user information and greater patient participation in the decision making process, as reflected by the law of March 4(th) 2002 pertaining to patients' rights and the quality of the healthcare system known as the Law on Democracy in healthcare. WHAT ABOUT TOOLS - DECISION SUPPORT FOR PATIENTS? At the micro level, some research projects are being developed, some of them using decision aids. Preliminary results show that patients want to be informed but that the concept of shared decision making needs to be analysed and refined from both the patients' and the physicians' points of views. WHAT ABOUT PROFESSIONAL INTEREST AND IMPLEMENTATION? However, the relationship between physicians/healthcare professionals and patients/healthcare users is very complex and progress in this field takes time. Only ten years after enactment of the Law on Democracy in healthcare, it might be premature to try and determine the state of the art of shared medical decision making at the macro and meso levels in France. WHAT DOES THE FUTURE LOOK LIKE? There is room in France for further studies on shared decision making in the medical encounter. Researchers, decision makers, healthcare users and healthcare professionals need a place to meet and exchange. An observatory dedicated to shared decision making will be launched in the coming months, both at the national level and in collaboration with several other French-speaking areas like Switzerland and the province of Quebec. PMID- 21620319 TI - Patient participation and shared decision making in Germany - history, agents and current transfer to practice. AB - The main focus of the present paper is to describe 1) the healthcare system specific influences on patient participation in medical decision making and 2) the current state of research and implementation of shared decision making (SDM) after ten years of substantial advances in health policy and research in this field. WHAT ABOUT POLICY REGARDING SDM? The "Medical Patients Rights Act" is to standardise all the rights and responsibilities within the scope of medical treatment. This also comprises the right to informed decisions, comprehensive and comprehensible information for patients, and decisions based on the partnership of clinicians and patients. WHAT ABOUT TOOLS - DECISION SUPPORT FOR PATIENTS? SDM training programmes for healthcare professionals have been developed and partly implemented. Several decision support interventions - primarily with support from health insurance funds - have been developed and evaluated. WHAT ABOUT PROFESSIONAL INTEREST AND IMPLEMENTATION? Against the background of the German health policy's endorsement of patient participation, the German government and other public institutions are currently funding different research programmes in which shared decision making is playing a substantial role. The development and implementation of decision support tools for patients and professionals as well as the implementation of trainings for healthcare professionals require stronger efforts. WHAT DOES THE FUTURE LOOK LIKE? With the support of health policy and with the utilisation of scientific evidence, the transfer of shared decision making into practice is considered to be meaningful in the German healthcare system. The translation into routine care will remain an important task for the future. PMID- 21620320 TI - The status of shared decision making and citizen participation in Israeli medicine. AB - WHAT ABOUT POLICY REGARDING SDM? Though informed consent and patients' right to information are regulated by Israeli law, there is a low level of formal activities focused on shared decision making (SDM) in Israel. Further, there are few organized programs to promote SDM among medical professionals or the public, and governmental support of SDM-related research is minimal. WHAT ABOUT TOOLS - DECISION SUPPORT FOR PATIENTS? The Israeli government does not have a program on development of patient decision aids. WHAT ABOUT PROFESSIONAL INTEREST AND IMPLEMENTATION? Nonetheless, patients have begun to influence litigation in both formal and informal capacities, medical schools have begun to incorporate courses for improving physician-patient communication into their curricula, and the largest national health plan has initiated a plan to increase pubic awareness. Funding for researching and promoting SDM is not centrally allocated, and studies show that despite the positive effects of SDM, such an approach is infrequently applied in actual clinical practice, and initiatives to promote SDM (e.g., decision aids) are in their infancy. WHAT DOES THE FUTURE LOOK LIKE? In conclusion, though not actively promoting SDM at present, Israel, with its governmentally regulated universal coverage with good access to high-level services possesses all the requisite elements for rapid, widespread advances in SDM in future years. PMID- 21620321 TI - Participation of patients and citizens in healthcare decisions in Italy. AB - WHAT ABOUT POLICY REGARDING SDM? The Italian National Health Plan and many regional and local health authorities explicitly recognise the importance of patient/citizen participation. These official documents rarely mention the specific concept of SDM, but they use terms such as patient involvement, participation, and empowerment. Patient associations are actively involved in promoting patient/citizen participation, but still play a marginal role in the health debate compared to health professionals, clinicians and healthcare managers. WHAT ABOUT TOOLS - DECISION SUPPORT FOR PATIENTS? There are only a few examples of decision aids designed for patients according to SDM concepts. A critical point is the lack of specific tools for the evaluation of SDM processes. Exceptions include the Italian versions of the OPTION scale and the SDM-Q, used at the micro-level for the evaluation of SDM. What about professional interest and real implementation? Health professionals recognise that doctor-patient communication is an important area. Italian research in SDM has grown and improved. It is very promising, but still limited. Undergraduate and postgraduate courses of some medical schools include specific programs on doctor-patient communication skills, focusing also on promotion of patient participation. WHAT DOES THE FUTURE LOOK LIKE? In conclusion, there is room for improving the Italian efforts to implement SDM into practice at both the individual and public level. PMID- 21620322 TI - Shared decision making in the Netherlands, is the time ripe for nationwide, structural implementation? AB - WHAT ABOUT POLICY REGARDING SDM? The Dutch health care system has been reformed in 2006 to make it more patient-oriented and demand-driven. We shortly describe four strategies of this health care reform. Although research projects are now fully spread over the country, a coordinated research agenda on SDM is lacking. WHAT ABOUT TOOLS - DECISION SUPPORT FOR PATIENTS? The Dutch governmental healthcare internet portal for patients hosts 16 patient decision aids. WHAT ABOUT PROFESSIONAL INTEREST AND IMPLEMENTATION? There is quite a strong patient participation movement in the Netherlands, on macro and meso level. Limited effort, related to the local research projects has been put into training professionals in SDM skills. WHAT DOES THE FUTURE LOOK LIKE? We need concerted action on the level of educating health care professionals, empowering patients, making patient decision aids easily accessible, supporting the professionals in this new task, and measuring the process of SDM in performance indicators used in quality assurance. The Dutch Platform for SDM that will be launched in Maastricht in June 2011 is therefore a timely and relevant initiative. PMID- 21620323 TI - Shared decision making in Spain: current state and future perspectives. AB - In the last two decades there has been a growing recognition in the Spanish National Health System (NHS) of the importance of considering patients' values and preferences in clinical decisions. Patient participation in shared decision making (SDM) is gaining importance as a suitable approach to patient-health professional communication and decision making in Spain. In addition, the NHS is funding the development of patients' decision aids (PtDAs) for shared decision making (SDM) by Health Technology Assessment Agencies. However, the NHS has still not incorporated reforms in law that includes SDM and PtDAs as a key component of health care services and professional curricula, nor is there a standardised implementation of interventions to support decisions in routine care. Most patients are not very familiar with their rights to be kept informed and participate in their own health care decisions. Most professionals are not familiar with or educated about patients' rights to be kept informed and participate in health care decisions either. The future of SDM in Spain is promising. The next course of action should be to maintain the production and adaptation of high-quality PtDAs while at the same time reinforcing effective dissemination strategies among patients and training programmes for professionals focused on SDM. PMID- 21620324 TI - Shared decision making development in Switzerland: room for improvement! AB - In Switzerland there is a strong movement at a national policy level towards strengthening patient rights and patient involvement in health care decisions. Yet, there is no national programme promoting shared decision making. First decision support tools (prenatal diagnosis and screening) for the counselling process have been developed and implemented. Although Swiss doctors acknowledge that shared decision making is important, hierarchical structures and asymmetric physician-patient relationships are still prevailing. The last years have seen some promising activities regarding the training of medical students and the development of patient support programmes. Swiss direct democracy and the habit of consensual decision making and citizen involvement in general may provide a fertile ground for SDM development in the primary care setting. PMID- 21620325 TI - Implementing shared decision making in the UK. AB - WHAT ABOUT POLICY REGARDING SDM? SDM is on the national policy agenda and has been prioritised as part of the health reform bill currently going through the Houses of Parliament. The NHS Constitution emphasises patients' right to be involved in decisions and this is reinforced in standards set by professional regulators. WHAT ABOUT TOOLS - DECISION SUPPORT FOR PATIENTS? The UK governments have invested in patient information and a few decision aids are freely available on public websites. WHAT ABOUT PROFESSIONAL INTEREST AND IMPLEMENTATION? There is interest in SDM and in collaborative care planning, but this is not yet the norm in clinical practice and few clinicians have received training in the topic. Several programmes are under way to encourage implementation of SDM. WHAT DOES THE FUTURE LOOK LIKE? Future developments will depend on the extent of clinical commitment and on whether there are sufficient funds available to invest in the promotion of SDM at a time when health care resources are tightly constrained. PMID- 21620326 TI - Shared decision making in the United States: policy and implementation activity on multiple fronts. AB - Shared decision making in the United States has become an important element in health policy debates. The recently passed federal health care reform legislation includes several key provisions related to shared decision making (SDM) and patient decision support. Several states have passed or are considering legislation that incorporates SDM as a key component of improved health care provision. Research on SDM is funded by a range of public and private organizations. Non-profit, for-profit, academic and government organizations are developing decision support interventions for numerous conditions. Some interventions are publicly available; others are distributed to patients through health insurance and healthcare providers. A significant number of clinical implementation projects are underway to test and evaluate different ways of incorporating SDM and patient decision support into routine clinical care. Numerous professional organizations are advocating for SDM and social networking efforts are increasing their advocacy as well. Policy makers are intrigued by the potential of SDM to improve health care provision and potentially lower costs. The role of shared decision making in policy and practice will be part of the larger health care reform debate. PMID- 21620327 TI - Measurement of shared decision making - a review of instruments. AB - The last years have seen a clear move towards shared decision making (SDM) and increased patient involvement in many countries. However, as the field of SDM research is still relatively young, new instruments for the measurement of (shared) decision making (process, outcome and surrounding elements) are constantly being developed. Thus, the aims of this structured review were to give an update on current developments regarding the measurement in the field of SDM, as well as to give a short overview of published and unpublished instruments. We conducted an electronic literature search in PubMed and the Web of Science database, performed hand searches of relevant journals and contacted key authors in the field. We found eight scales that have been subjected to further psychometric testing, eleven new and psychometrically tested instruments and nine developments that are still in the publishing process. The results show that there is a trend towards measuring SDM processes from a dyadic approach (assessing both the patient's and the clinician's perspective). More and more scales have been developed and tested in languages other than English, which indicates the growing research efforts in various countries. While reliability of most scales is good, they differ in their extent of validation. Further psychometric testing is needed, as well as the development of a theoretical measurement framework in order to improve consistency of measured constructs across research groups. PMID- 21620328 TI - Sequence divergence at mitochondrial genes in animals: applicability of DNA data in genetics of speciation and molecular phylogenetics. AB - Nucleotide diversity estimates for the genes Cyt-b (cytochrome b) and Co-1 (cytochrome oxidase 1) are analyzed. Genetic divergence of populations (1) and taxa of different rank, such as subspecies, semispecies or/and sibling species (2), species within a genus (3), species from different genera within a family (4), and species from separate families within an order (5) have been compared using a database of p-distances and similar measures. Empirical data for 20,731 vertebrate and invertebrate animal species reveal various and increasing levels of genetic divergence of the sequences of the two genes, Cyt-b and Co-1, in the five groups compared. Mean unweighted scores of p-distances (%) for five groups are: Cyt-b (1) 1.38+/-0.30, (2) 5.10+/-0.91, (3) 10.31+/-0.93, (4) 17.86+/-1.36, (5) 26.36+/-3.88 and Co-1 (1) 0.89+/-0.16, (2) 3.78+/-1.18, (3) 11.06+/-0.53, (4) 16.60+/-0.69, (5) 20.57+/-0.40. These estimates testify to the applicability of p distance for most intraspecies and interspecies comparisons of genetic divergence up to the order level for the two genes compared. The results of the analysis of the nucleotide divergence within species and higher taxa of animals suggest that a phyletic evolution in animals is likely to prevail at the molecular level, and speciation mainly corresponds to the geographic or divergence mode (type D1). The prevalence of the D1 speciation mode does not mean that other modes are absent. At least seven possible modes of speciation are considered. The approach suggested that allows recognize the speciation modes formally with the operational genetic criteria. Such approach may help to solve a key problem of the biological species concept, i.e. the lack of ability to monitor in most cases the reproductive isolation barriers between species. PMID- 21620329 TI - In silico mining and characterization of simple sequence repeats from gilthead sea bream (Sparus aurata) expressed sequence tags (EST-SSRs); PCR amplification, polymorphism evaluation and multiplexing and cross-species assays. AB - We screened for simple sequence repeats (SSRs) found in ESTs derived from an EST database development project ('Marine Genomics Europe' Network of Excellence). Different motifs of di-, tri-, tetra-, penta- and hexanucleotide SSRs were evaluated for variation in length and position in the expressed sequences, relative abundance and distribution in gilthead sea bream (Sparus aurata). We found 899 ESTs that harbor 997 SSRs (4.94%). On average, one SSR was found per 2.95 kb of EST sequence and the dinucleotide SSRs are the most abundant accounting for 47.6% of the total number. EST-SSRs were used as template for primer design. 664 primer pairs could be successfully identified and a subset of 206 pairs of primers was synthesized, PCR-tested and visualized on ethidium bromide stained agarose gels. The main objective was to further assess the potential of EST-SSRs as informative markers and investigate their cross-species amplification in sixteen teleost fish species: seven sparid species and nine other species from different families. Approximately 78% of the primer pairs gave PCR products of expected size in gilthead sea bream, and as expected, the rate of successful amplification of sea bream EST-SSRs was higher in sparids, lower in other perciforms and even lower in species of the Clupeiform and Gadiform orders. We finally determined the polymorphism and the heterozygosity of 63 markers in a wild gilthead sea bream population; fifty-eight loci were found to be polymorphic with the expected heterozygosity and the number of alleles ranging from 0.089 to 0.946 and from 2 to 27, respectively. These tools and markers are expected to enhance the available genetic linkage map in gilthead sea bream, to assist comparative mapping and genome analyses for this species and further with other model fish species and finally to help advance genetic analysis for cultivated and wild populations and accelerate breeding programs. PMID- 21620330 TI - A transcriptomic scan for positively selected genes in two closely related marine fishes: Sebastes caurinus and S. rastrelliger. AB - Comparative genomic analyses can provide valuable insight into functional evolutionary divergence among closely related species. Here we employ a comparative evolutionary analysis of expressed sequence tags (ESTs) from two closely related species of marine fishes (genus Sebastes--rockfish). Sebastes is a highly diverse group of marine fishes that inhabit a wide array of marine habitats and the study of this group can provide insights into speciation in the marine environment. ESTs were developed for S. caurinus (23,668 from brain, kidney, and spleen tissues) and S. rastrelliger (11,207 from brain and pituitary tissues). Following assembly we were able to identify, with high confidence, 257 orthologous sequence pairs between the two species through a reciprocal best hit blast search. An analysis of functional divergence between orthologs revealed that 19.46% had Ka/Ks values greater than 0.5 and 8.17% had Ka/Ks values greater than one, identifying a large pool of candidate genes to further study adaptive divergence in the group. Genes with elevated Ka/Ks values belonged to the following functional categories: immune function, metabolism, longevity, and reproductive behavior, indicating that adaptive divergence in these functional groups may be important in the diversification of this group of fishes. This study provides the ground work to better understand the molecular evolution of genes involved in a radiation of marine fishes. PMID- 21620331 TI - Molecular cloning of CYP4 and GSTpi homologues in the scallop Chlamys farreri and its expression in response to benzo[a]pyrene exposure. AB - Cytochrome P450 enzymes (CYP) and glutathione s-transferases (GST) are essential components of cellular detoxification systems. In this study we cloned full length cDNAs encoding CYP4 and GSTpi homologues from scallop Chlamys farreri. Both sequences were deposited in the GenBank with accession no. ACL80141 for CYP4 and ACL80138 for GSTpi. The sequence called Cf (C. farreri) CYP4 is constituted by an ORF of 1317 bp encoding for a protein of 50.8 kDa. The CfGSTpi is constituted by an ORF of 618 bp encoding for a protein of 23.9 kDa. The comparison of the deduced amino acid sequences with CYP4 and GSTpi from vertebrates showed high conservation of the residues and domains essential to the function of these two enzymes. CfCYP4 and CfGSTpi mRNA expression was detected in digestive gland, gill, mantle, mature female gonad and adductor. We then utilized the real-time PCR to study expression levels of the CfCYP4 and CfGSTpi gene in response to exposure of Benzo[a]pyrene (BaP) (0.01 and 0.2 MUg/L) for 10 days. The results showed that during the exposure to BaP, CfCYP4 was significantly decreased in the gill and digestive gland of scallops, and CfGSTpi was increased on day 3 until the end of exposure. The changes in CfGSTpi mRNA levels observed in scallops exposed to BaP indicated that GSTpi could play an important role in the detoxification of BaP. PMID- 21620332 TI - Comparative analysis of intronless genes in teleost fish genomes: insights into their evolution and molecular function. AB - This study assessed the relationship between the occurrence and function of intronless or single exon genes (SEG) in the genome of five teleost species and their phylogenetic distance. The results revealed that Takifugu rubripes, Tetraodon nigroviridis, Oryzias latipes, Gasterosteus aculeatus and Danio rerio genomes are respectively comprised of 2.83%, 3.42%, 4.49%, 4.35% and 4.02% SEGs. These SEGs encode for a variety of family proteins including claudins, olfactory receptors and histones that are essential for various biological functions. Subsequently, we predicted and annotated SEGs in three European sea bass, Dicentrarchus labrax chromosomes that we have sequenced, and compared results with those of stickleback (G. aculeatus) homologous chromosomes. While the annotation features of three D. labrax chromosomes revealed 78 (5.30%) intronless genes, comparisons with G. aculeatus showed that SEG composition and their order varied significantly among corresponding chromosomes, even for those with nearly complete synteny. More than half of SEGs identified in most of the species have at least one ortholog multiple exon gene in the same genome, which provides insight to their possible origin by retrotransposition. In spite of the fact that they belong to the same lineage, the fraction of predicted SEGs varied significantly between the genomes analyzed, and only a low fraction of proteins (4.1%) is conserved between all five species. Furthermore, the inter-specific distribution of SEGs as well as the functional categories shared by species did not reflect their phylogenetic relationships. These results indicate that new SEGs are continuously and independently generated after species divergence over evolutionary time as evidenced by the phylogenetic results of single exon claudins genes. Although the origin of SEGs cannot be inferred directly from the phylogeny, our results provide strong support for the idea that retrotransposition followed by tandem duplications is the most probable event that can explain the expansion of SEGs in eukaryotic organisms. PMID- 21620333 TI - Molecular cloning, characterization and expression analysis of thrombospondin gene from Penaeus monodon. AB - In present study, a thrombospondin gene was obtained from the ovary and neurosecretory organ in eyestalk cDNA library of black tiger prawn (Penaeus monodon). The full-length P. monodon thrombospondin (PmTSP) cDNA contained a 5' untranslated region (UTR) of 9 bp, an open reading frame (ORF) of 2778 bp encoding a polypeptide of 925 amino acids with molecular mass 100.57 kDa, and a 3'UTR of 99 bp. ScanProsite analysis indicated that PmTSP contained four chitin binding type-II domains, an EGF-like domain, eight thrombospondin type-III repeats and one thrombospondin C-terminal domain. Homology analysis of the deduced amino acid sequence of the PmTSP with other known TSP sequences by MatGAT software revealed that the PmTSP shows very high homology with the sequences of Fennerpenaeus chinensis (89.9% similarity, 83.8% identity). Analysis of the tissue expression pattern of the PmTSP gene showed that the PmTSP mRNA was expressed in all tested tissues, including hepatopancreas, ovary, muscle, intestine, neurosecretory organ in eyestalk, neurosecretory organ in brain, stomach, and heart, with highest level in the ovary. Furthermore, the PmTSP expression was found to be of high level in six development stages of the ovary. The results indicated that PmTSP might play an important role in ovarian development. PMID- 21620334 TI - Transcriptome sequencing of a highly salt tolerant mangrove species Sonneratia alba using Illumina platform. AB - Mangroves are critical and threatened marine resources, yet few transcriptomic and genomic data are available in public databases. The transcriptome of a highly salt tolerant mangrove species, Sonneratia alba, was sequenced using the Illumina Genome Analyzer in this study. Over 15 million 75-bp paired-end reads were assembled into 30,628 unique sequences with an average length of 581 bp. Of them, 2358 SSRs were detected, with di-nucleotide repeats (59.2%) and tri-nucleotide repeats (37.7%) being the most common. Analysis of codon usage bias based on 20,945 coding sequences indicated that genes of S. alba were less biased than those of some microorganisms and Drosophila and that codon usage variation in S. alba was due primarily to compositional mutation bias, while translational selection has a relatively weak effect. Genome-wide gene ontology (GO) assignments showed that S. alba shared a similar GO slim classification with Arabidopsis thaliana. High percentages of sequences assigned to GO slim category 'mitochondrion' and four KEGG pathways, such as carbohydrates and secondary metabolites metabolism, may contribute to salt adaptation of S. alba. In addition, 1266 unique sequences matched to 273 known salt responsive genes (gene families) in other species were screened as candidates for salt tolerance of S. alba, and some of these genes showed fairly high coverage depth. At last, we identified four genes with signals of strong diversifying selection (K(a)/K(s)>1) by comparing the transcriptome sequences of S. alba with 249 known ESTs from its congener S. caseolaris. This study demonstrated a successful application of the Illumina platform to de novo assembly of the transcriptome of a non-model organism. Abundant SSR markers, salt responsive genes and four genes with signature of natural selection obtained from S. alba provide abundant sequence sources for future genetic diversity, salt adaptation and speciation studies. PMID- 21620335 TI - Regional analgesia and acute pain management. Foreword. PMID- 21620336 TI - Regional analgesia and acute pain management: major leaps in small steps? Preface. PMID- 21620337 TI - Ultrasound-guided regional anesthesia for peripheral nerve blocks: an evidence based outcome review. AB - Ultrasound-guided regional anesthesia (UGRA) has increased in popularity over the past 5 years. This interest is reflected by the plethora of publications devoted to technique development, as well as randomized and controlled trials. Despite the excitement around ultrasonography, skeptics argue that there is a lack of evidence-based medicine to support the unequivocal adoption of UGRA as a "standard of care." This article summarizes and critically assesses current data comparing traditional approaches to localizing nerves with those that use ultrasound guidance. In addition, the potential benefits of UGRA that go beyond current information available from comparative studies are explored. PMID- 21620338 TI - Continuous peripheral nerve blocks in the hospital and at home. AB - A single-injection peripheral nerve block using long-acting local anesthetic provides analgesia for 12 to 24 hours; however, many surgical procedures result in pain that lasts far longer. One relatively new option is a continuous peripheral nerve block (CPNB): local anesthetic is perfused via a perineural catheter directly adjacent to the peripheral nerve(s) supplying the surgical site, providing potent, site-specific analgesia. CPNB results in decreased pain, opioid requirements, opioid-related side effects, and sleep disturbances; in some cases, accelerating resumption of tolerated passive joint range-of-motion and increasing patient satisfaction. Ambulatory perineural infusion may be provided using a portable infusion pump, in some cases resulting in decreased hospitalization duration and related costs. Serious complications are rare, but may result in significant morbidity. PMID- 21620339 TI - Economics and practice management issues associated with acute pain management. AB - The use of regional anesthesia (RA) improves cost benefit (hospital-centered) and cost utility (patient-centered) over general anesthesia with volatile agents (GAVA), based upon research in outpatient populations. To make the cost savings a reality, the authors recommend: (1) avoidance of GAVA or at least volatile agents, (2) adopting published postanesthesia care unit (PACU)-bypass criteria conducive to RA, (3) maximizing PACU-bypass rates, and (4) utilizing a block induction area. Inpatient-based acute pain services are not uniform, which makes cost analyses and comparison between practices unreliable. Additional review and commentary address surgical site infections, cancer recurrence, blood transfusions, and chronic postsurgical pain. PMID- 21620340 TI - Local anesthetic systemic toxicity: prevention and treatment. AB - Anesthesia is a sine qua non for most surgeries. Like any medical advance, progress in regional anesthesia has not come without its share of complications, including a spectrum extending from localized nerve injury to systemic cardiovascular toxicity and death. This article discusses the mechanisms and clinical presentation, prevention, treatment, and future trends of local anesthetic systemic toxicity. The adverse effects of lipid emulsion therapy are also included. PMID- 21620341 TI - Assessment and treatment of postblock neurologic injury. AB - The incidence of neurologic damage after regional anesthesia is rare. However, this complication may have dramatic consequences for the patient because recovery may take several months. As nerve conduction studies and electromyography are the cornerstones of investigations in cases of postblock deficit, it is mandatory for the anesthesiologist performing regional anesthesia to have a basic understanding of these tests to discuss the cause with the surgeon and inform the patient about the prognosis. PMID- 21620342 TI - Complications of regional anesthesia and acute pain management. AB - Perioperative nerve injuries are recognized as a complication of regional anesthesia. Although rare, studies suggest the frequency of complications is increasing. Risk factors include neural, traumatic injury during needle or catheter placement, infection, and choice of local anesthetic solution. Neurologic injury due to pressure from improper patient positioning, tightly applied casts or surgical dressings, and surgical trauma are often attributed to regional anesthetic. Body habitus and preexisting neurologic dysfunction may also contribute. The safe conduct of regional anesthesia involves knowledge of patient, anesthetic, and surgical risk factors. Early diagnosis and treatment of reversible etiologies are critical to optimizing neurologic outcome. PMID- 21620343 TI - Unintentional subdural injection: a complication of neuraxial anesthesia/analgesia. AB - Unintentional subdural injection during neuraxial anesthesia/analgesia continues to be a challenge for anesthesiologists. This unusual complication is often poorly recognized, with the diagnosis made in retrospect, or not at all. The clinical presentation of these regional blocks can be heterogeneous, ranging from restricted, patchy, or unilateral sensory blockade all the way to extensive and even life-threatening motor and autonomic nervous system depression. Prompt diagnosis using clinical algorithms and radiographic imaging is crucial for the early discontinuation of the offending catheter. Supportive care is mandatory in cases involving severe depression of consciousness, motor function, and/or sympathetic tone. PMID- 21620344 TI - Challenges in acute pain management. AB - The management of acute pain remains challenging, with many patients suffering inadequate pain control following surgery. Certain populations are at unique risk for unrelieved pain. Evidence-based approaches taking into account patients' specific needs and problems will likely substantially improve their perioperative experience. These patients must be identified in the preoperative process, and an anesthetic/analgesic plan discussed and formulated. A targeted multimodal approach to pain management should be considered the best clinical practice. The most challenging patients may benefit most from the surveillance of an acute pain service that is able to monitor and coordinate care into the postoperative period. PMID- 21620345 TI - New concepts in acute pain management: strategies to prevent chronic postsurgical pain, opioid-induced hyperalgesia, and outcome measures. AB - Chronic postsurgical pain (CPSP) is a pain syndrome that has attracted attention for more than 10 years. CPSP is a pain syndrome that develops postoperatively and lasts for at least 2 months in the absence of other causes for pain (eg, recurrence of malignancy, chronic infection, and so forth). Pain continuing from a preexisting disease is not considered as CPSP. In this article, the authors discuss the etiopathogenesis of CPSP and interventions that can help prevent and treat this condition. PMID- 21620346 TI - Local infiltration analgesia. AB - Pain after major abdominal, orthopedic, and thoracic surgeries can be significant causing unacceptable morbidity. Poorly controlled pain results in patient dissatisfaction and may also be associated with major morbidities, including perioperative myocardial ischemia, pulmonary complications, altered immune function, and postoperative cognitive dysfunction. Various techniques are currently used to manage this pain, and opioids are amongst the most frequently used. Recent literature supports the use of regional anesthesia in the form of various peripheral nerve blocks as a better alternative. This article discusses the role and evidence for wound infiltration analgesia in general surgery, orthopedic surgery, neurosurgery, and thoracic surgery. PMID- 21620347 TI - [Crateriform nodule on the helix of the ear]. PMID- 21620348 TI - [Validation of the Spanish Version of the Itch Severity Scale: the PSEDA study]. AB - BACKGROUND AND OBJECTIVES: The Itch Severity Scale (ISS) facilitates objective assessment of the intensity of pruritus. The aim of this study was to validate a Spanish version of the ISS in patients with atopic dermatitis. MATERIAL AND METHODS: A prospective epidemiological study was undertaken in patients diagnosed with atopic dermatitis at least 1 year previously and a control group without the disease. Patients with atopic dermatitis were stratified according to the status of the lesions (active or inactive) and questionnaires were completed at baseline and 3-month and 6-month follow-up. Data were collected on sociodemographic variables relating to atopic dermatitis (including the modified Eczema Area and Severity Index [mEASI]), concomitant disease, and patient measures such as ISS, Dermatology Life Quality Index (DLQI), and Children's DLQI (cDLQI). RESULTS: A total of 207 children (2-17 years) were included: 56 control subjects, 103 patients with active lesions, and 48 with inactive lesions. The mean (SD) age of the participants in this age group was 8.1 (4.0) years. A total of 261 adults (>= 18 years) were included: 89 control subjects, 124 patients with active lesions, and 48 with inactive lesions. The mean age of the adult participants was 32.3 (13.4) years. A response rate of > 80% was obtained on the pediatric ISS (feasibility) and the responses correlated with the mEASI and cDLQI at baseline (P<.001) as an indicator of validity. An effect size of 0.988 was observed (sensitivity to change) along with a Chronbach alpha of 0.840 (internal consistency). A response rate of >95% was obtained on the adult ISS (feasibility) and the responses correlated with the mEASI and DLQI at baseline (P<.001) as an indicator of validity. An effect size of 1.0 was observed (sensitivity to change) along with a Chronbach alpha of 0.825 (internal consistency). CONCLUSIONS: The Spanish version of the ISS is feasible, valid, sensitive to change, and displays good reliability based on internal consistency in both children and adults. PMID- 21620350 TI - [Vitamin D: evidence and controversies]. AB - Vitamin D enhances musculoskeletal health and reduces mortality related to bone disease in some populations, particularly the elderly and other high-risk groups. Evidence suggests that vitamin D has an impact in cancer, cardiovascular disease, autoimmune processes, and infections. Epidemiologic studies have also detected vitamin D deficits or insufficiency in nearly all the world's populations. Such evidence has led to debate related, to a certain degree, to photoprotective measures that aim at protecting against skin cancer. The latest recommendations of the American Institute of Medicine consider that serum levels of 20 ng/mL (50 nmol/L) appear to be adequate in the general population and achievable even with minimal sun exposure. If these figures are reliable, the apparent pandemic of vitamin D deficiency reported in recent years may be exaggerated. This article reviews the evidence and issues under discussion, looking especially at the role ultraviolet radiation plays in synthesizing vitamin D in the skin. The conclusion is that sun exposure should not be used as the only source of vitamin D given that it is also clearly carcinogenic for the skin. A healthful approach combines moderate sun exposure, adequate food sources of the vitamin, and supplements whenever required. PMID- 21620351 TI - Concentration polarization effect at the deposition of charged Langmuir monolayers. AB - The review summarizes the results of the recent studies of the electrokinetic relaxation process within the meniscus region during the deposition of charged Langmuir monolayers. Such electrokinetic relaxation is the consequence of the initial misbalance of partial ion fluxes within a small region near the contact line, where the diffuse parts of electric double layers, formed at the monolayer and the substrate surface, overlap. The concentration polarization within the solution near the three-phase contact line should lead to long-term relaxations of the meniscus after beginning and stopping the deposition process, to changes of the ionic composition within the deposited films, to change of the interaction of the monolayer with the substrate, and to dependence of the maximum deposition rate on the subphase composition. PMID- 21620352 TI - DASH: a method for identical-by-descent haplotype mapping uncovers association with recent variation. AB - Rare variants affecting phenotype pose a unique challenge for human genetics. Although genome-wide association studies have successfully detected many common causal variants, they are underpowered in identifying disease variants that are too rare or population-specific to be imputed from a general reference panel and thus are poorly represented on commercial SNP arrays. We set out to overcome these challenges and detect association between disease and rare alleles using SNP arrays by relying on long stretches of genomic sharing that are identical by descent. We have developed an algorithm, DASH, which builds upon pairwise identical-by-descent shared segments to infer clusters of individuals likely to be sharing a single haplotype. DASH constructs a graph with nodes representing individuals and links on the basis of such segments spanning a locus and uses an iterative minimum cut algorithm to identify densely connected components. We have applied DASH to simulated data and diverse GWAS data sets by constructing haplotype clusters and testing them for association. In simulations we show this approach to be significantly more powerful than single-marker testing in an isolated population that is from Kosrae, Federated States of Micronesia and has abundant IBD, and we provide orthogonal information for rare, recent variants in the outbred Wellcome Trust Case-Control Consortium (WTCCC) data. In both cohorts, we identified a number of haplotype associations, five such loci in the WTCCC data and ten in the isolated, that were conditionally significant beyond any individual nearby markers. We have replicated one of these loci in an independent European cohort and identified putative structural changes in low-pass whole genome sequence of the cluster carriers. PMID- 21620353 TI - Adaptor protein complex 4 deficiency causes severe autosomal-recessive intellectual disability, progressive spastic paraplegia, shy character, and short stature. AB - Intellectual disability inherited in an autosomal-recessive fashion represents an important fraction of severe cognitive-dysfunction disorders. Yet, the extreme heterogeneity of these conditions markedly hampers gene identification. Here, we report on eight affected individuals who were from three consanguineous families and presented with severe intellectual disability, absent speech, shy character, stereotypic laughter, muscular hypotonia that progressed to spastic paraplegia, microcephaly, foot deformity, decreased muscle mass of the lower limbs, inability to walk, and growth retardation. Using a combination of autozygosity mapping and either Sanger sequencing of candidate genes or next-generation exome sequencing, we identified one mutation in each of three genes encoding adaptor protein complex 4 (AP4) subunits: a nonsense mutation in AP4S1 (NM_007077.3: c.124C>T, p.Arg42(*)), a frameshift mutation in AP4B1 (NM_006594.2: c.487_488insTAT, p.Glu163_Ser739delinsVal), and a splice mutation in AP4E1 (NM_007347.3: c.542+1_542+4delGTAA, r.421_542del, p.Glu181Glyfs(*)20). Adaptor protein complexes (AP1-4) are ubiquitously expressed, evolutionarily conserved heterotetrameric complexes that mediate different types of vesicle formation and the selection of cargo molecules for inclusion into these vesicles. Interestingly, two mutations affecting AP4M1 and AP4E1 have recently been found to cause cerebral palsy associated with severe intellectual disability. Combined with previous observations, these results support the hypothesis that AP4-complex mediated trafficking plays a crucial role in brain development and functioning and demonstrate the existence of a clinically recognizable syndrome due to deficiency of the AP4 complex. PMID- 21620354 TI - Mutations in the N-terminal actin-binding domain of filamin C cause a distal myopathy. AB - Linkage analysis of the dominant distal myopathy we previously identified in a large Australian family demonstrated one significant linkage region located on chromosome 7 and encompassing 18.6 Mbp and 151 genes. The strongest candidate gene was FLNC because filamin C, the encoded protein, is muscle-specific and associated with myofibrillar myopathy. Sequencing of FLNC cDNA identified a c.752T>C (p.Met251Thr) mutation in the N-terminal actin-binding domain (ABD); this mutation segregated with the disease and was absent in 200 controls. We identified an Italian family with the same phenotype and found a c.577G>A (p.Ala193Thr) filamin C ABD mutation that segregated with the disease. Filamin C ABD mutations have not been described, although filamin A and filamin B ABD mutations cause multiple musculoskeletal disorders. The distal myopathy phenotype and muscle pathology in the two families differ from myofibrillar myopathies caused by filamin C rod and dimerization domain mutations because of the distinct involvement of hand muscles and lack of pathological protein aggregation. Thus, like the position of FLNA and B mutations, the position of the FLNC mutation determines disease phenotype. The two filamin C ABD mutations increase actin binding affinity in a manner similar to filamin A and filamin B ABD mutations. Cell-culture expression of the c.752T>C (p.Met251)Thr mutant filamin C ABD demonstrated reduced nuclear localization as did mutant filamin A and filamin B ABDs. Expression of both filamin C ABD mutants as full-length proteins induced increased aggregation of filamin. We conclude filamin C ABD mutations cause a recognizable distal myopathy, most likely through increased actin affinity, similar to the pathological mechanism of filamin A and filamin B ABD mutations. PMID- 21620355 TI - Optimal timing of periodontal disease treatment for prevention of adverse pregnancy outcomes: before or during pregnancy? AB - Several large randomized controlled clinical trials failed to find that standard periodontal therapy during pregnancy reduces the incidence of adverse pregnancy outcomes (eg, preterm birth and low birthweight). However, treating periodontal disease during pregnancy may be too late to reduce the inflammation that is related to the adverse pregnancy outcomes. Moreover, periodontal treatment during pregnancy can cause bacteremia, which itself may initiate the pathway leading to the adverse pregnancy outcomes. Finally, the periodontal treatments provided during pregnancy are not always effective in preventing the progression of periodontal disease during pregnancy. Pregnancy may not be an appropriate period for periodontal intervention(s). We hypothesize that periodontal treatment before pregnancy may reduce the rates of adverse pregnancy outcomes. Future randomized controlled trials are needed to test if treating periodontal disease in the prepregnancy period reduces the rate of adverse pregnancy outcomes. PMID- 21620356 TI - Barriers to pelvic floor physical therapy utilization for treatment of female urinary incontinence. AB - OBJECTIVE: The purpose of this study was to estimate the effect of insurance status on pelvic floor physical therapy (PFPT) nonparticipation for the treatment of urinary incontinence. STUDY DESIGN: A cross-sectional study of women referred to PFPT for urinary incontinence between January 2009 and June 2010 was conducted. A telephone questionnaire was administered. Multiple logistic regression was used to identify risk factors for nonparticipation. RESULTS: Thirty-three percent of women with private insurance and 17% with other insurance were PFPT nonparticipants. On multiple logistic regression, women with Medicare were more likely to participate in PFPT (odds ratio [OR], 0.12; 95% confidence interval [CI], 0.01-0.72). Risk factors for nonparticipation included insurance noncoverage (OR, 103.85; 95% CI, 6.21-infinity) and a negative perception regarding the benefit of PFPT (OR, 5.07; 95% CI, 2.16-12.49). CONCLUSION: Among women who were referred to PFPT for urinary incontinence, insurance noncoverage and negative patient perception of efficacy were risk factors for nonparticipation, although having Medicare was protective. Improving patient education and insurance coverage for PFPT may increase usage. PMID- 21620358 TI - Relationship between the time interval from antenatal corticosteroid administration until preterm birth and the occurrence of respiratory morbidity. AB - OBJECTIVE: The purpose of this study was to assess the relationship between neonatal respiratory morbidity and the interval between antenatal corticosteroids (ACS) administration and birth. STUDY DESIGN: We performed a retrospective cohort study among women who had received ACS and delivered at <34 weeks of gestation. We categorized these women in 4 groups: ACS-to-delivery interval of 0-7, 8-14, 15 21, and 22-28 days. Multivariable logistic regression analysis assessed the association between the ACS-to-delivery interval and neonatal respiratory morbidity. RESULTS: We included 254 neonates. Eighty-two neonates (32%) were intubated. In comparison with neonates with an ACS-to-delivery interval of 0-7 days, the risk for intubation was increased in all other groups (odds ratio [OR], 2.3; 95% confidence interval [CI], 1.1-5.4; OR, 5.6; 95% CI, 1.8-18; and OR, 4.8; 95% CI, 0.71-32, not statistically significant, respectively). CONCLUSION: The effect of ACS decreases when the ACS-to-delivery interval exceeds 7 days. The first administration of ACS should be considered carefully. PMID- 21620357 TI - Pharmacokinetics of 17-hydroxyprogesterone caproate in multifetal gestation. AB - OBJECTIVE: The purpose of this study was to define the pharmacokinetic parameters of 17-hydroxyprogesterone caproate (17-OHPC) in multifetal gestation. STUDY DESIGN: Blood was obtained at 24-28 weeks' gestation and at 32-35 weeks gestation in 97 women with twin and 26 women with triplet gestation who were receiving 17 OHPC. Six of the women with twins had daily blood sampling for 7 days between 24 and 28 weeks' gestation, and pharmacokinetic parameters were estimated with the use of noncompartmental analysis. Modeling was applied to estimate the population parameters and to simulate various treatment scenarios. RESULTS: The apparent half-life of 17-OHPC was 10 days. Body mass index significantly impacted 17-OHPC concentrations, but fetal number and parity did not. Apparent clearance was significantly greater in African American than in white women (P = .025). CONCLUSION: This is the first pharmacokinetic analysis of 17-OHPC in pregnant women. Determination of half-life, covariates that affect plasma 17-OHPC concentrations, and the modeling of drug behavior provide insights into this drug's pharmacologic properties during multifetal pregnancy. PMID- 21620359 TI - Levator avulsion using a tomographic ultrasound and magnetic resonance-based model. AB - OBJECTIVE: Delivery-related levator avulsion can cause pelvic floor dysfunction. We compared agreement between tomographic ultrasound and magnetic resonance-based models for the detection of levator defects. STUDY DESIGN: Sixty-nine Chinese women with pelvic organ prolapse were assessed prospectively by 3-dimensional ultrasound scans and magnetic resonance imaging. Levator-urethra gap (LUG), levator-symphysis gap (LSG), and puborectalis attachment width were measured offline with state-of-the-art software. Interobserver variability and agreement between the 2 methods were determined. RESULTS: Interobserver repeatability was moderate-to-excellent for all parameters that were measured with both methods and agreement between methods in diagnosing levator avulsion. LUG and LSG measurements were significantly higher in women with a levator avulsion. With a diagnosis of complete levator avulsion, receiver operating characteristics analysis suggested a cutoff of 23.65 mm for LUG and 28.7 mm for LSG. CONCLUSION: Levator avulsion can be diagnosed reliably by tomographic ultrasound scanning and magnetic resonance imaging evaluation, and linear measures, such as LSG and LUG, can be proxy measurements for avulsion. PMID- 21620360 TI - Vaginal cuff closure after minimally invasive hysterectomy: our experience and systematic review of the literature. AB - OBJECTIVE: To determine the incidence of vaginal cuff dehiscence after minimally invasive hysterectomy, we reported our series of total laparoscopic hysterectomies with transvaginal colporraphy. STUDY DESIGN: We then conducted a systematic search of PubMed to retrieve published series of laparoscopic and robotic hysterectomies, in which different techniques for vaginal cuff closure were used. RESULTS: In our study group, vaginal cuff dehiscence occurred in 2 of 665 (0.3%) patients. Our literature search identified 57 articles, for a total of 13,030 endoscopic hysterectomies. Ninety-one postoperative vaginal separations were reported (0.66%). The pooled incidence of vaginal dehiscence was lower for transvaginal cuff closure (0.18%) than for both laparoscopic (0.64%; odds ratio [OR], 0.28; 95% confidence interval [CI], 0.12-0.65) and robotic (1.64%; OR, 0.11; 95% CI, 0.04-0.26) colporraphy. Laparoscopic cuff closure was associated with a lower risk of dehiscence than robotic closure (OR, 0.38; 95% CI, 0.28 0.6). CONCLUSION: Current evidence indicates that transvaginal colporraphy after total laparoscopic hysterectomy is associated with a 3- and 9-fold reduction in risk of vaginal cuff dehiscence compared with laparoscopic and robotic suture, respectively. PMID- 21620361 TI - Pregravid cardiometabolic risk profile and risk for gestational diabetes mellitus. AB - OBJECTIVE: The purpose of this study was to examine pregravid cardiometabolic profile and subsequent risk of gestational diabetes mellitus (GDM). STUDY DESIGN: GDM cases (n = 199) and control subjects (n = 381) were selected from a cohort of women who took part in a multiphasic health checkup examination at Kaiser Permanente from 1984-1996 and who had a subsequent pregnancy and were matched by year and age at multiphasic health checkup examination and age at delivery. RESULTS: Pregravid measurements of serum glucose levels of 100-140 mg/dL, body mass index of >= 25.0 kg/m(2), and prehypertension/hypertension level were associated independently with GDM risk (odds ratios [OR], 4.8; 95% confidence interval [CI], 1.7-13.9; OR, 2.7; 95% CI, 1.6-4.3; and OR, 1.5; 95% CI, 1.0-2.3, respectively). The risk of GDM increased with the number of cardiometabolic risk factors (P < .001); adverse levels of both body mass index and glucose were associated with a 4.6-fold increased risk of GDM, compared with women with normal levels (P = .0001). CONCLUSION: Pregravid cardiometabolic risk profile may help clinicians to identify high-risk women to target for primary prevention or early management of GDM. PMID- 21620362 TI - Tissue Doppler echocardiographic markers of cardiac dysfunction in small-for gestational age fetuses. AB - OBJECTIVE: The objective of the study was to evaluate echocardiographic markers of cardiac dysfunction in small-for-gestational age (SGA) fetuses with normal umbilical artery Doppler. STUDY DESIGN: Cardiac function was evaluated in 58 SGA (mean gestational age, 38 weeks) and 58 gestational-age matched normally grown fetuses by conventional echocardiography (peak early [E] and late [A] ratios and myocardial performance index [MPI]), and tissue Doppler imaging (TDI) (annular peak velocities and MPI'). RESULTS: With conventional echocardiography, SGA fetuses had a nonsignificant trend to increased E/A ratios and left MPI compared with controls. TDI demonstrated that SGA fetuses had significantly lower right E' and A' peak velocities and higher MPI' values. CONCLUSION: These findings further support that a proportion of SGA fetuses have true late-onset intrauterine growth restriction, which is associated with subclinical cardiac dysfunction, as previously described for early-onset intrauterine growth restriction. PMID- 21620363 TI - Could chronic pelvic pain be a functional somatic syndrome? AB - The cause of noncyclical chronic pelvic pain (CPP) in many women is unknown: 30% have no identifiable pelvic pathology, and in those who do the relationship of CPP and the pathology is often unclear. Moreover, epidemiologic studies demonstrate that the common findings of endometriosis and adhesions do not greatly increase the odds of having CPP. CPP and the functional somatic syndromes (fibromyalgia, irritable bowel syndrome, and others) share many characteristics including pain as a prominent symptom and comorbidity. For the functional somatic syndromes, the initial focus of etiologic investigations has been on local mechanisms and then on systemic pathogeneses. We believe that the research trajectories of the functional somatic syndromes and CPP are converging. Their juncture might reveal an important pathologic mechanism for CPP in some women that is primarily outside the pelvis. This observation would open up new areas of exploration and treatment of CPP. PMID- 21620364 TI - Is an episiotomy necessary with a shoulder dystocia? AB - OBJECTIVE: The objective of the study was to determine whether a decrease in the use of episiotomy was associated with a change in the frequency of brachial plexus injury. STUDY DESIGN: All births at Brigham and Women's Hospital from Sept. 1, 1998, through Aug. 31, 2009, were reviewed. The total number of births, mode of delivery, shoulder dystocias, episiotomies with and without shoulder dystocias, and brachial plexus injuries were recorded. A nonparametric test of trend was performed. RESULTS: There were a total of 94,842 births, 953 shoulder dystocias, and 102 brachial plexus injuries. The rate of episiotomy with shoulder dystocia dropped from 40% in 1999 to 4% in 2009 (P = .005) with no change in the rate of brachial plexus injuries per 1000 vaginal births. CONCLUSION: Despite historical recommendations for an episiotomy to prevent brachial plexus injury when a shoulder dystocia is encountered, the trend we observed does not suggest benefit from this practice. PMID- 21620366 TI - Why do some pregnant women prefer cesarean? The influence of parity, delivery experiences, and fear. AB - OBJECTIVE: We sought to identify predictors of preferences for cesarean among pregnant women, and estimate how different predictors influence preferences. STUDY DESIGN: This was a cross-sectional study based on the Norwegian Mother and Child Cohort Study (n = 58,881). RESULTS: Of the study population, 6% preferred cesarean over vaginal delivery. While 2.4% of nulliparous had a strong preference for cesarean, the proportion among multiparous was 5.1%. The probability that a woman, absent potential predictors, would have a cesarean preference was similar (<2%) for both nulliparous or multiparous. In the presence of concurrent predictors such as previous cesarean, negative delivery experience, and fear of birth, the predicted probability of a cesarean request ranged from 20-75%. CONCLUSION: The proportion of women with a strong preference for cesarean was higher among multiparous than nulliparous women, but the difference was attributable to factors such as previous cesarean or fear of delivery and not to parity per se. PMID- 21620365 TI - Adolescent pregnancy and gestational weight gain: do the Institute of Medicine recommendations apply? AB - OBJECTIVE: The purpose of this study was to examine the Institute of Medicine (IOM) guidelines for gestational weight gain in adolescents. STUDY DESIGN: We studied a retrospective cohort using the Missouri Birth Certificate Registry and included subjects who were primiparous, who had singleton gestations, who were <20 years old, and who delivered at 24-44 weeks gestation. The exposure was defined as weight gain less than, within, or greater than IOM recommendations. Outcomes that were examined were small-for-gestational-age (SGA) infants, large for-gestational age (LGA) infants, preterm delivery, infant death, preeclampsia, cesarean delivery, and operative vaginal delivery. The analysis was stratified by body mass index category. RESULTS: In any body mass index category, inadequate weight gain was associated with increased odds of SGA infants, preterm delivery, and infant death. When subjects gained more than the IOM recommendations, the number of SGA infants decreased, with slight increases in the number of LGA infants, preeclampsia, and cesarean delivery. CONCLUSION: Adolescents should be counseled regarding adequate weight gain in pregnancy. Further research is necessary to determine whether the IOM recommendations recommend enough weight gain in adolescents to optimize pregnancy outcomes. PMID- 21620367 TI - Anti--vascular endothelial growth factor therapy in glaucoma filtration surgery. AB - PURPOSE: To examine evidence supporting the use of vascular endothelial growth factor (VEGF) inhibition in controlling wound healing after glaucoma filtration surgery in primary open-angle glaucoma, to identify the optimum method of administration, and to clarify the potential position of anti-VEGF monoclonal antibodies in comparison with 5-fluorouracil and mitomycin C. DESIGN: Perspective based on an overview of evidence from current peer-reviewed literature. METHODS: Analysis of evidence from animal studies, in vitro studies, human studies, and from the use of anti-VEGF monoclonal antibodies in systemic disease. RESULTS: There is evidence that glaucoma patients have elevated levels of VEGF in the aqueous before filtration surgery, that this increases in animals after filtration surgery, and that both can be suppressed in animals by intraocular injection of bevacizumab. VEGF not only has a role in angiogenesis, but also has a direct action on fibroblast activity that may be modified directly at the time of filtration surgery. CONCLUSIONS: There is evidence for a role for VEGF in wound healing after glaucoma filtration surgery. The optimum route of administration and dosing regimen of anti-VEGF antibodies and their positioning in comparison with 5-fluorouracil and mitomycin C are uncertain. There is some evidence that subconjunctival injection may produce sustained intraocular tissue levels. There is also evidence that bevacizumab may act in synergy with 5 fluorouracil. Although there are no direct comparative studies, it seems unlikely that bevacizumab alone will be as effective as mitomycin C, although bleb morphologic features may be better. PMID- 21620368 TI - [Protein C deficiency in black African with venous thromboembolism in Cotonou, Benin]. AB - The aim of this study is to evaluate the frequency of protein C deficiency in venous thromboembolism in black African patients of Benin. It is a descriptive study. Inclusion criteria were: acceptance- having a venous thromboembolism. No exlusion criteria was retained. Protein C deficiency was diagnosed by quantitative technic with a Minividas materiel in the blood. Protein C dosage has been done before antivitamin k therapy and a second dosage has been done if the first one demonstrated a low level of protein C. Acuired aetiology have been research. For the 54 patients of this study mean age was 52.7+/-14.1 and sex ratio 1.08. The frequency of protein C deficiency was 9.3% in all patients and 12.5% in those with clinical thrombophily (p=1). No acquired deficit has been found. PMID- 21620369 TI - One and a half ventricle repair in adults: postoperative hemodynamic assessment using phase-contrast magnetic resonance imaging. AB - BACKGROUND: One and a half ventricle repair (11/2 repair) strategy has been used for patients with a hypoplastic or dysfunctional right ventricle (RV), or both. We sought to assess the postoperative hemodynamics of 11/2 repair using phase contrast magnetic resonance imaging (PC-MRI). METHODS: Ten adults, 9 with Ebstein's anomaly and 1 with tricuspid stenosis, underwent 11/2 repair (median age at operation, 42.4 years). The azygos vein was left open in all patients on 11/2 repair to prevent severe postoperative central venous hypertension. Postoperative PC-MRI studies were performed to measure blood flow to the ascending aorta (QAsc-Ao), the main pulmonary artery (QMPA), the superior vena cava (SVC) (QSVC), and the branch pulmonary arteries and veins. From these values, blood flow to the upper compartment of the body (QUC), right ventricular volume unloading effect (QMPA/QAsc-Ao), proportion of blood flow to the upper compartment of the body (QUC/QAsc-Ao), and venous return to arterial forward flow ratio of the upper compartment of the body (QSVC/QUC) were calculated. Two patients also underwent preoperative PC-MRI. RESULTS: On PC-MRI, QMPA/QAsc-Ao, QUC/QAsc-Ao, and QSVC/QUC were 0.58 to 0.84 (median, 0.67), 0.19 to 0.36 (median, 0.27), and 0.47 to 1.57 (median, 0.93, lower than 1.0 in 7 patients), respectively. In 2 patients who had preoperative and postoperative PC-MRI, QUC/QAsc-Ao decreased from 0.26, 0.32 to 0.21, 0.28, respectively. CONCLUSIONS: After 11/2 repair, right ventricular volume unloading was effective in all patients, but intercaval collateral veins (ie. QSVC/QUC<1) appeared to develop in most of the patients. Furthermore blood flow to the upper compartment of the body appeared to diminish, presumably due to postoperative elevation of central venous pressure. PMID- 21620370 TI - Ross root dilation in adult patients: is preoperative aortic insufficiency associated with increased late autograft reoperation? AB - BACKGROUND: The Ross aortic valve replacement (AVR) offers excellent hemodynamic and clinical outcomes in most patients undergoing aortic root replacement. Because of ongoing debate regarding the durability of this procedure, long-term follow-up in a large adult Ross experience might be of interest. METHODS: Between 1994 and 2010, 230 adult patients underwent modified Ross root procedures. Mean age was 42.4+/-12.1 years (range, 20 to 68 years), 62% were male. Fifty-six patients (24%) had undergone one or more prior aortic valve interventions. Concomitant procedures were performed in 126 patients (55%), including 90 ascending aortic procedures. Presenting pathology was aortic insufficiency in 114 (50%) and aortic stenosis in 116 (50%). RESULTS: Hospital mortality or within 30 days was 2 of 230 patients (0.9%). During follow-up (mean, 7.8 years), 12 more patients died. At 15 years, survival was 94%, and freedom from autograft and allograft reoperation was 91% and 98%, respectively. Reoperation was required for 23 patients. Eighteen patients required reoperation on the autograft root; 11 of 18 (61%) had preoperative aortic insufficiency as their predominate aortic valve lesion. Freedom from autograft reoperation was not significantly different for patients with preoperative aortic insufficiency (87%) compared with patients who had aortic stenosis (94%; p=0.15). On multivariable analysis, no risk factors were significant for reoperation except for surgery before 2000 (p<0.0001) and previous AVR (p=0.05). CONCLUSIONS: Preoperative aortic regurgitation was not a significant risk factor for late autograft reoperation in adults. The Ross AVR provides excellent hemodynamics and survival for adults willing to accept a small risk of reoperation as opposed to a life-long risk of thromboemboli and anticoagulation therapy as exist with mechanical aortic prostheses. Reoperation risk for Ross AVR has decreased with Ross annulus sinotubular junction reinforcement replacement of a dilated ascending aorta and postoperative hypertension management. PMID- 21620371 TI - Current evidence for perioperative statins in cardiac surgery. AB - Cardiac surgery improves life expectancy and quality of life for the constantly ageing population in developed countries. Mediated by their lipid-dependent and lipid-independent mechanisms, statins are sought to provide benefit with regard to better outcomes after cardiac surgery. Current guidelines recommend statin use in patients undergoing coronary artery bypass grafting, while less evidence is available for patients referred to heart valve surgery. Optimal selection of statin drug and dosage including perioperative timing of statin therapy remains largely unknown, but results of ongoing meta-analyses and future randomized trials will add important evidence to guide perioperative statin treatment of cardiac surgery patients. PMID- 21620372 TI - Neoadjuvant chemoradiation for clinically advanced non-small cell lung cancer: an analysis of 233 patients. AB - BACKGROUND: Surgical intervention after chemoradiation for locoregionally advanced non-small cell lung cancer (NSCLC) is controversial. This study evaluated patient survival after neoadjuvant chemoradiation and anatomic pulmonary resections for locoregionally advanced NSCLC. METHODS: Clinicopathologic data were retrospectively collected for 233 patients (110 women, 123 men) with NSCLC who underwent chemoradiation therapy, followed by pneumonectomy, sleeve lobectomy, bilobectomy, and standard lobectomy, from 1989 to 2008. Univariate log-rank analysis of Kaplan-Meier survival curves and multivariate Cox regression analysis was performed. RESULTS: Final pathologic stages were complete responders, 52 (22%); I, 56 (24%); II, 39 (17%); and III, 86 (37%). Final pathologic lymph node status was N0, 130 (56%); N1, 28 (12%); and N2, 75 (32%). Overall 5-year survival for the cohort was 43%. The 90-day mortality was 8% (18 of 233). The 5-year survival was 33% for pneumectomy vs 51% for lobectomy (p=0.002). Survival rates at 5 years by stage were complete responders, 58%; I, 50%; II, 41%; and III, 32%; by primary tumor status, T0, 50%; T2, 38%; T3, 29%; and T4, 28%; and by final pathologic nodal status, N0, 51%; N1, 40%; N2, 32% (N0 vs N1, p=0.236; N1 vs N2, p=0.704; N0 vs N2, p=0.019; N0 vs N1+N2, p=0.020). Multivariate analysis demonstrated pneumonectomy was associated with decreased 5-year survival (hazard risk, 1.5162; 95% confidence interval, 10.05028 to 2.189, p=0.0263). CONCLUSIONS: Respectable survival can be achieved after neoadjuvant chemoradiation, followed by anatomic resection, in selected patients with clinically advanced NSCLC. A T0 primary tumor or N0 lymph node status individually, or together as a complete response (T0 N0) status, is associated with the best long-term survival. Survival is most favorable for lobectomies vs pneumonectomies after neoadjuvant chemoradiation therapy. PMID- 21620373 TI - Postnatal expression of sialin in the mouse submandibular gland. AB - OBJECTIVE: Sialin has been identified as a sialic acid and aspartate/glutamate transporter. Both cytoplasmic localization and the plasma membrane labelling pattern suggested that sialin may possess multiple transport functions in different cell types. In mouse embryos, sialin expression was primarily detected in the central nervous system. However, sialin shows widespread and high-level expression in adult tissues. Despite its ubiquitous expression and important functions, the postnatal expression profile and subcellular localization of sialin in the salivary gland remains elusive. The aim of the present study was to investigate the expression and subcellular distribution of sialin during postnatal development in the mouse submandibular gland (SMG). DESIGN: Six SMGs from both female and male C57BL/6 mice were collected at P10, P30 and P90, and the material from each littermate of either sex was pooled to extract total RNA and tissue protein. The remaining tissues were immediately fixed in 10% neutral buffered formalin for histological analysis. The mRNA and protein expression levels of sialin were examined by quantitative real-time RT-PCR and Western blot analysis. The subcellular distribution of sialin was analysed by immunohistochemistry and immunofluorescence. RESULTS: The postnatal expression level of sialin in the mouse SMG was comparable with that in brain at each time point tested. The temporal expression of sialin in the SMG gradually increased during postnatal maturation. Immunohistochemical and immunofluorescence analysis demonstrated that sialin was predominantly expressed on the basal cytoplasmic membrane of acini and ducts, as well as in some myoepithelial cells in the SMG. CONCLUSIONS: The high-level expression and subcellular distribution pattern of sialin in the SMG suggest that sialin may play an important role in the transport and secretion of saliva. PMID- 21620374 TI - Corticospinal excitability is dependent on the parameters of peripheral electric stimulation: a preliminary study. AB - OBJECTIVE: To evaluate the effect of 6 electric stimulation paradigms on corticospinal excitability. DESIGN: Using a same subject pre-post test design, transcranial magnetic stimulation (TMS) was used to measure the responsiveness of corticomotor pathway to biceps and triceps brachii muscles before and after 30 minutes of electric stimulation over the biceps brachii. Six different electric stimulation paradigms were applied in random order, at least 3 days apart. SETTING: Motor control research laboratory. PARTICIPANTS: Healthy subjects (N=10; 5 women, 5 men; mean age +/- SD, 26 +/- 3.6y). INTERVENTIONS: Six different electric stimulation paradigms with varied stimulus amplitude, frequency, and ramp settings. MAIN OUTCOME MEASURE: Amplitudes of TMS-induced motor evoked potentials at biceps and triceps brachii normalized to maximal M-wave amplitudes. RESULTS: Electric stimulation delivered at stimulus amplitude sufficient to evoke a sensory response at both 10 Hz and 100 Hz, and stimulus amplitude to create a noxious response at 10 Hz decreased corticomotor responsiveness (all P<0.01). Stimulation sufficient to induce a motor contraction (30 Hz) applied in a ramped pattern to mimic a voluntary activation increased corticomotor responsiveness (P=0.002), whereas constant low- and high-intensity motor stimulation at 10 Hz did not. Corticomotor excitability changes were similar for both the stimulated muscle and its antagonist. CONCLUSIONS: Stimulus amplitude (intensity) and the nature (muscle flicker vs contraction) of motor stimulation have a significant impact on changes in corticospinal excitability induced by electric stimulation. Here, we demonstrate that peripheral electric stimulation at stimulus amplitude to create a sensory response reduces corticomotor responsiveness. Conversely, stimulus amplitude to create a motor response increases corticomotor responsiveness, but only the parameters that create a motor response that mimics a voluntary muscle contraction. PMID- 21620375 TI - Hyperbaric oxygen therapy in the management of paroxysmal sympathetic hyperactivity after severe traumatic brain injury: a report of 6 cases. AB - Paroxysmal sympathetic hyperactivity (PSH) after severe brain injury is detrimental to the recovery of patients. Pharmacologic management of PSH is difficult and efficacy is unpredictable or incomplete. This report presents 6 cases of PSH after extremely severe traumatic brain injury in which hyperbaric oxygen therapy (HBOT) controlled paroxysmal autonomic changes and posturing in the early subacute phase after limited success with conventional medication regimens. Thus, HBOT may present an option for the management of PSH in addition to pharmacologic therapy. Potential mechanisms for these effects are discussed. PMID- 21620376 TI - Attenuating memory distrust in a repeated checking task. AB - Repeated checking has been demonstrated to lead to reductions in memory confidence in several previous studies using student and clinical samples. This process of reduced confidence in memory and detail for memory, are thought to arise from the inhibition of perceptual processing that develops during repeated checking. Our research investigated whether reduced memory confidence from repeated checking could be attenuated through the use of novel stimuli during the repeated checking task. Three groups were generated through random assignment of 65 undergraduate students. As seen in previous research, individuals who repeatedly checked a stimulus (a virtual stovetop) showed reduced memory confidence, vividness, and detail, when compared with individuals who repeatedly checked a different stimulus. A third group in which the colour of the repeatedly checked stovetop changed every five trials showed no significant decline in memory confidence between the pre-test and post-test. Results suggest that increased memory distrust can be ameliorated through the use of stimuli with characteristics that are novel and distinctive. Findings are discussed in the context of the existing model of repeated checking and memory confidence, and implications for treatment methods are presented. PMID- 21620377 TI - Effect of cognitive load on eye-target synchronization during smooth pursuit eye movement. AB - In mild traumatic brain injury (mTBI), the fiber tracts that connect the frontal cortex with the cerebellum may suffer shear damage, leading to attention deficits and performance variability. This damage also disrupts the enhancement of eye target synchronization that can be affected by cognitive load when subjects are tested using a concurrent eye-tracking test and word-recall test. We investigated the effect of cognitive load on eye-target synchronization in normal and mTBI patients using the nonlinear dynamical technique of stochastic phase synchronization. Results demonstrate that eye-target synchronization was negatively affected by cognitive load in mTBI subjects. In contrast, eye-target synchronization improved under intermediate cognitive load in young (<=40years old) normal subjects. PMID- 21620378 TI - Vasoactive intestinal peptide protects against ischemic brain damage induced by focal cerebral ischemia in rats. AB - Vasoactive intestinal peptide (VIP) exerts neuroprotective effects under various neurotoxic conditions in vitro. In the present study, we investigated the effects of VIP on transient ischemic brain damage. Focal cerebral ischemia was induced using middle cerebral artery occlusion (MCAO) for 120 min in the adult rat brain. Either a single intracerebroventricular injection of VIP or saline was given at the beginning of reperfusion. Forty-eight hours after MCAO, the rats were sacrificed for evaluation of the infarct volume and histological analysis. ELISA was performed to assay levels of serum S100B before being sacrificed. We also evaluated the blood-brain barrier (BBB) permeability using Evans blue dye injection method. In contrast to the cases treated with vehicle, the infarct volume was significantly (P<0.05) reduced, and terminal deoxynucleotidyl transferase-mediated dUTP-nick end labeling (TUNEL) staining and immunoreactivity for S100B were also significantly (P<0.05) decreased in the ischemic hemisphere with VIP treatment. In addition, the elevations of serum S100B were significantly (P<0.01) attenuated in VIP-treated rats compared with those of control rats. Treatment with VIP did not result in a significant reduction of Evans blue leakage, although it tended to be lower than that in the control rats. Our data suggest that treatment with VIP reduces brain damage in ischemic rats, and this effect may be associated with the attenuation of apoptosis and S100B expression. PMID- 21620379 TI - Immunohistochemical localization of transient receptor potential vanilloid type 1 and insulin receptor substrate 2 and their co-localization with liver-related neurons in the hypothalamus and brainstem. AB - The central nervous system plays an important role in the regulation of energy balance and glucose homeostasis mainly via controlling the autonomic output to the visceral organs. The autonomic output is regulated by hormones and nutrients to maintain adequate energy and glucose homeostasis. Insulin action is mediated via insulin receptors (IR) resulting in phosphorylation of insulin receptor substrates (IRS) inducing activation of downstream pathways. Furthermore, insulin enhances transient receptor potential vanilloid type 1 (TRPV1) mediated currents. Activation of the TRPV1 receptor increases excitatory neurotransmitter release in autonomic centers of the brain, thereby impacting energy and glucose homeostasis. The aim of this study is to determine co-expression of IRS2 and TRPV1 receptors in the paraventricular nucleus of the hypothalamus (PVN) and dorsal motor nucleus of the vagus (DMV) in the mouse brain as well as expression of IRS2 and TRPV1 receptors at liver-related preautonomic neurons pre-labeled with a trans-neural, viral tracer (PRV-152). The data indicate that IRS2 and TRPV1 receptors are present and co-express in the PVN and the DMV. A large portion (over 50%) of the liver-related preautonomic DMV and PVN neurons expresses IRS2. Moreover, the majority of liver-related DMV and PVN neurons also express TRPV1 receptors, suggesting that insulin and TRPV1 actions may affect liver-related preautonomic neurons. PMID- 21620380 TI - Gender differences in hyperthermia and regional 5-HT and 5-HIAA depletion in the brain following MDMA administration in rats. AB - In the present research the role of gender in MDMA-induced hyperthermia and serotonin depletion is studied by injecting male and female male rats with MDMA or saline 3 times (i.p.) with 3h interval at dosages of 0.3, 1, 3 or 9 mg/kg at an ambient temperature of 25 degrees C. The acute hyperthermia following the higher dosages was much stronger in males than in females. After the highest dose, body temperature was even raised for several days. This effect was particularly present in males where nocturnal hyperthermia persisted the whole 4 week period of sampling. Despite the differences in the acute hyperthermic response, no significant gender differences were found in 5-HT depletion 4 weeks after MDMA (9 mg/kg) administration. A striking difference was present, however, in the concentration of the 5-HT metabolite 5-HIAA after MDMA administration. In males 5-HIAA levels decreased, whereas in females this metabolite was hardly affected, suggesting a lasting increase in 5-HT turnover in females following drug administration. When genders were matched for their acute physiological hyperthermic response by repeated injection of 9 mg/kg in female rats and 6 mg/kg in male rats, 5-HT depletion was only present in females. In this experiment with matched acute physiological responses 5-HIAA levels also decreased much stronger in males, suggesting an increased 5-HT turnover in females 4 weeks after MDMA administration. In conclusion, although male rats are clearly more susceptible for the acute as well as the lasting hyperthermic effects of MDMA than females, this is not reflected in levels of 5-HT depletion following administration of similar dosages of the drug. This may indicate that, in case of a similar thermogenic response, females have a higher 5-HT neurotoxicity following MDMA than males. PMID- 21620381 TI - MG132 enhances neurite outgrowth in neurons overexpressing mutant TAR DNA-binding protein-43 via increase of HO-1. AB - In patients with amyotrophic lateral sclerosis (ALS), various mutations were identified in TAR DNA-binding protein-43 (TDP-43). In the present study, we found that mutant TDP-43 inhibited the neurite outgrowth. Subsequently, we tested the effect of MG132 on the mutant TDP-43 cell lines. Non-toxic doses of MG132 promoted neurite extension and decreased the level of thiobarbituric acid reactive substances (TBARS). Heme oxygenase-1 (HO-1) known as antioxidase was restored by MG132. Conversely, Zinc protoporphyrin IX (ZnPP IX), which is an inhibitor of heme oxygenase, inhibited neurite outgrowth induced by MG132. It was well known that HO-1 was regulated by nuclear factor E2-related factor 2 (Nrf2). However, MG132 increased the expression of HO-1 independent of the Nrf2 pathway. PMID- 21620382 TI - Reactivity switching and selective activation of C-1 or C-3 in 2,3-unsaturated thioglycosides. AB - Reactivity switching and selective activation of C-1 or C-3 in 2,3-unsaturated thioglycosides, namely, 2,3-dideoxy-1-thio-D-hex-2-enopyranosides are reported. The reactivity switching allowed activation of either C-1 or C-3, with the use of either N-iodosuccinimide (NIS)/triflic acid (TfOH) or TfOH alone. C-1 glycosylation with alcohol acceptors occurred in the presence of NIS/TfOH, without the acceptors reacting at C-3. On the other hand, reaction of 2,3 unsaturated thioglycosides with alcohols mediated by triflic acid led to transposition of C-1 ethylthio-moiety to C-3 intramolecularly, to form 3 ethylthio-glycals. Resulting glycals underwent glycosylation with alcohols to afford 3-ethylthio-2-deoxy glycosides. However, when thiol was used as an acceptor, only a stereoselective addition at C-3 resulted, so as to form C-1, C-3 dithio-substituted 2-deoxypyranosides. PMID- 21620383 TI - Furfural formation from d-xylose: the use of different halides in dilute aqueous acidic solutions allows for exceptionally high yields. AB - Starting from the results achieved in a previous work on the effects of Cl(-) ions on furfural formation in aqueous acid solution [Marcotullio, G. et al., Green Chem.2010, 12, 1739], the general effect of different halides is addressed. Experimental results show the halides to influence at least two distinct steps in the reaction leading from d-xylose to furfural under acidic conditions, via different mechanisms. The nucleophilicity of the halides appears to be critical for the dehydration, but not for the initial enolization reaction. By combining different halides synergic effects become evident resulting in very high selectivities and furfural yields. PMID- 21620384 TI - Carbasugar analogues of galactofuranosides: beta-O-linked derivatives and towards beta-S-linked derivatives. AB - A selectively protected carbasugar analogue of beta-galactofuranose was synthesised from glucose using ring-closing metathesis as the key step. The carbasugar was converted into an alpha-galacto configured 1,2-epoxide, which was an effective electrophile in Lewis acid catalysed coupling reactions with alcohols. The epoxide was opened with regioselective attack at C-1 to give beta galacto configured C-1 ethers. Using carbohydrates as nucleophiles, we synthesised a number of pseudodisaccharides. The epoxide was also regioselectively opened at C-1 with a sulfur nucleophile under basic conditions to give a beta-galacto configured C-1 thioether. PMID- 21620385 TI - CCL5 induces a pro-inflammatory profile in microglia in vitro. AB - The chemokine receptors CCR1, CCR2, CCR3, CCR5, and CXCR2 have been found to be expressed on microglia in many neurodegenerative diseases, such as multiple sclerosis and Alzheimer's disease. There is emerging evidence that chemokines, besides chemoattraction, might directly modulate reactive profiles of microglia. To address this hypothesis we have investigated the effects of CCL2, CCL3, CCL5, and CXCL1 on cytokine and growth factor production, NO synthesis, and phagocytosis in non-stimulated and lipopolysaccharide-stimulated primary rat microglia. The respective receptors CCR1, CCR5, and CXCR2 were shown to be functionally expressed on microglia. All tested chemokines stimulated chemotaxis whereas only CCL5 increased NO secretion and attenuated IL-10 as well as IGF-1 production in activated microglia. Based on these findings we propose that besides its chemoattractant function CCL5 has a modulatory effect on activated microglia. PMID- 21620386 TI - A pain in the neck--imaging in neck sepsis. AB - Deep neck infection has a high morbidity and mortality and the extent of infection is often difficult to estimate clinically. The complex anatomy and the communication between neck spaces means that infection can spread along fascial planes leading to life-threatening complications such as airway compromise, vascular erosion/thrombosis, neural dysfunction, and ultimately descending necrotizing mediastinitis. Imaging has an important role to play in identifying the extent of infection and the presence of complications. PMID- 21620387 TI - Emotional eating in anorexia nervosa and bulimia nervosa. AB - OBJECTIVES: The relationship between emotional states and eating behaviors is complex, and emotional eating has been identified as a possible factor triggering binge eating in bulimia nervosa (BN) and binge eating disorder. Few studies considered emotional eating in patients with anorexia nervosa. METHODS: The present study evaluated the clinical correlates of emotional eating in 251 eating disordered (EDs) subjects (70 AN restricting type, 71 AN binge eating/purging type, 110 BN purging type) and in a group of 89 healthy control subjects. Subjects were assessed by means of a clinical interview (Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition) and several self-reported questionnaires, including the Emotional Eating Scale (EES). RESULTS: No significant differences were found between the 3 EDs groups in terms of EES total score, and all patients with ED showed higher EES scores compared with control subjects. Emotional eating was associated with subjective binge eating in AN binge eating/purging type and with objective binge eating in patients with BN. Among patients with AN restricting type, emotional eating was associated with restraint, but this association was lost when controlling for fear of loss of control over eating, which was the principal determinant of restraint. CONCLUSION: Emotional eating and fear of loss of control over eating are significantly associated with specific eating attitudes and behaviors, according to the different diagnoses. Emotional eating is a relevant psychopathologic dimension that deserves a careful investigation in both anorectic and bulimic patients. PMID- 21620388 TI - Privacy and governance implications of wider societal uses of brain imaging data. PMID- 21620389 TI - Reading others' minds by measuring their brains: fascinating and challenging for science, but ready for use in court? PMID- 21620390 TI - Brain imaging: consent issues, and licensing of storage and analysis. PMID- 21620391 TI - [Anemia in chronic heart failure]. AB - Anemia is one of the most common comorbidities in patients with decompensated chronic heart failure admitted to the Internal Medicine Ward. However, although there is evidence supporting its treatment to improve the functional capacity of the patients and to reduce the new admissions rate, the clinical practice guidelines do not provide any directives regarding its approach. This is an ideal clinical problem for the internist due to its multifactorial origin and the comprehensive point of view needed to approach the group of syndromes that occur in these patients (anemia, heart failure, geriatric syndromes, diabetes, etc.) The choice of treatment strategy, if such treatment is decided, should always begin after correcting the congestive signs in the outpatient with optimal treatment of heart failure. PMID- 21620392 TI - Bowel resection for iatrogenic parasitic fibroids with preoperative investigations suggestive of malignancy. AB - OBJECTIVE: To describe a case in which a midline laparotomy for two presumed malignant masses instead revealed parasitic fibroids. DESIGN: Case report. SETTING: Tertiary-level private hospital. PATIENT(S): Woman with 7- and 13-cm abdominopelvic masses 3 years after a total laparoscopic hysterectomy for fibroids. INTERVENTION(S): Midline laparotomy, bilateral salpingooophorectomy, infracolic omentectomy, appendicectomy, para-aortic lymph node dissection, and high rectosigmoid resection with primary anastomosis. MAIN OUTCOME MEASURE(S): Histology of masses showed adenomyoma. RESULT(S): There was no evidence of malignancy. CONCLUSION(S): Morcellation of the fibroids during a total laparoscopic hysterectomy likely left fragments that formed iatrogenic parasitic fibroids, which led to the subsequent laparotomy and bowel resection for potential malignancy. PMID- 21620393 TI - Nationwide use of postmortem retrieved sperm in Israel: a follow-up report. AB - A follow-up study of the subsequent use of all postmortem frozen sperm samples during 2003-2010 is reported. Only the sister of one of the 10 unmarried deceased men was in contact with the bank. Four widows elected to discard the frozen sperm and all of the remaining spouses were uninterested in its fate. Because none of the samples were requested for use, the need for sperm procurement should be reconsidered. PMID- 21620394 TI - Saliva cortisol levels and subjective stress are not associated with number of oocytes after controlled ovarian hyperstimulation in patients undergoing in vitro fertilization. AB - OBJECTIVE: To establish an association between the number of oocytes retrieved after controlled ovarian hyperstimulation (COH) and saliva cortisol (F) levels, as well as subjective stress, in women undergoing their first IVF cycle. DESIGN: Prospective clinical study. SETTING: Academic research institution. PATIENT(S): Women with primary or secondary infertility undergoing IVF. INTERVENTION(S): Fertility problem inventory (FPI) questionnaire and measurement of morning and evening saliva F by RIA. MAIN OUTCOME MEASURE(S): Number of oocytes and stress, defined as low morning F and/or a positive FPI result. RESULT(S): Eighty-three women provided saliva specimens, 66 of whom also filled in the FPI. The median number of oocytes was 8.4 (range 0-26). A state of stress was observed in 38/83 (46%) women. The mean number of oocytes was not significantly different between women with and without stress (7.3 +/- 4.3 vs. 8.9 +/- 6.9, respectively). In a multivariate analysis, stress (odds ratio 2.6; 95% confidence interval 0.03 225.7) and morning F (odds ratio 0.9; 95% confidence interval 0.6-1.3) were not significantly associated with the number of oocytes. There were no statistically significant correlations between F concentrations, FPI results, and age, number of poor responders, live birth rate, and clinical pregnancy rate (PR). CONCLUSION(S): Stress, as measured by saliva F and the FPI questionnaire, does not negatively impact the effectiveness of COH and is not associated with a reduced number of oocytes. PMID- 21620396 TI - A "conditionally essential" nutrient, L-carnitine, as a primary suspect in endometriosis. AB - L-carnitine, when administered to young female mice, has been shown to induce a pathologic condition resembling human endometriosis accompanied by a marked degree of infertility. Thus, the use of this nutrient by young women may be a potential risk factor responsible for the onset of endometriosis at a later stage of their lives. PMID- 21620395 TI - Effects of oral contraceptives on natriuretic peptide levels in women with hypothalamic amenorrhea: a pilot study. AB - Natriuretic peptides, which are important regulators of salt handling and blood pressure, are 60%-75% higher in healthy young women than in men, consistent with a gender dimorphism. In this randomized, placebo-controlled study in women with functional hypothalamic amenorrhea, we show that administration of oral contraceptives (OC) increases natriuretic peptide levels and that end-of-study free T levels are inversely associated with amino-terminal pro-B-type natriuretic peptide levels, consistent with the hypothesis that natriuretic peptide levels may be mediated by differences in gonadal steroid concentrations-estrogens (E) or androgens. PMID- 21620397 TI - Semen and hormonal parameters in men with chronic hepatitis C infection. AB - Male patients with chronic hepatitis C virus (HCV) infection (n = 57) demonstrated a statistically significant decrease in semen volume, sperm count, and progressive sperm motility and a statistically significant increase in abnormal sperm morphology compared with healthy controls (n = 40). The duration of the HCV infection was negatively correlated with semen volume and sperm motility where the HCV RNA viral load was negatively correlated with sperm count and sperm motility. Chronic HCV patients had statistically significantly lower total serum testosterone and higher serum E(2) and prolactin levels compared with healthy controls. PMID- 21620398 TI - Case series of duodenal follicular lymphoma, observed by magnified endoscopy with narrow-band imaging. PMID- 21620399 TI - EUS-guided fistulization of postoperative colorectal stenosis in an infant with Hirschsprung's disease: a new technique. PMID- 21620400 TI - Touch preparation of jumbo forceps biopsies allows rapid adequacy assessment of subepithelial GI masses. PMID- 21620401 TI - Indications, detectability, positive findings, total enteroscopy, and complications of diagnostic double-balloon endoscopy: a systematic review of data over the first decade of use. AB - BACKGROUND: Double-balloon endoscopy (DBE) has been used in clinical practice for nearly 10 years. OBJECTIVE: To systematically collect and produce pooled data on indications, detection rate, total enteroscopy, complications, and the composition of positive findings in diagnostic DBE. DESIGN: A systematic review. MAIN OUTCOME MEASUREMENTS: We searched PubMed between January 1, 2001 and March 31, 2010 for original articles about DBE evaluation of small-bowel diseases. Data on total number of procedures, distribution of indications, pooled detection rate, pooled total enteroscopy rate, and composition of positive findings were extracted and/or calculated. In addition, the data involving DBE-associated complications were analyzed. RESULTS: A total of 66 English-language original articles involving 12,823 procedures were included. Suspected mid-GI bleeding (MGIB) was the most common indication (62.5%), followed by symptoms/signs only (7.9%), small-bowel obstruction (5.8%), and Crohn's disease (5.8%). The pooled detection rates were 68.1%, 68.0%, 53.6%, 63.4%, and 85.8% for overall, suspected MGIB, symptoms/signs only, Crohn's disease, and small-bowel obstruction, respectively. Inflammatory lesions (37.6%) and vascular lesions (65.9%) were the most common findings, respectively, in suspected MGIB patients of Eastern and Western countries. The pooled total enteroscopy rate was 44.0% by combined or antegrade-only approach. The pooled minor and major complication rates were 9.1% and 0.72%, respectively. LIMITATIONS: Inclusion and exclusion criteria were loosely defined. CONCLUSION: The detectability and complication risk of diagnostic DBE are acceptable. Suspected MGIB is the most common indication, with a relatively high detection rate, but there was a difference in its causes between Western and Eastern countries. PMID- 21620402 TI - History of sex trafficking, recent experiences of violence, and HIV vulnerability among female sex workers in coastal Andhra Pradesh, India. AB - OBJECTIVES: To estimate the prevalence of sex trafficking as a mode of entry into sex work, and to examine associations between sex trafficking and recent violence experiences and HIV vulnerability among female sex workers (FSWs). METHODS: In a cross-sectional study in 2006 in coastal Andhra Pradesh, India, 812 FSWs were recruited via respondent-driven sampling to take part in an oral survey of their experiences in sex work. RESULTS: One in 5 (19.3%) FSWs met the UN definition of sex trafficking. Women trafficked into sex work were more likely than other FSWs to report recent violence experiences (adjusted odds ratio [AOR], 1.93; 95% confidence interval [CI], 1.32-2.81), more clients per week (AOR, 1.63; 95% CI, 1.11-2.41), and more days of sex work per week (AOR, 1.76; 95% CI, 1.18-2.63), and were less likely to report use of FSW-focused services (AOR, 0.60; 95% CI, 0.42-0.86). No significant differences emerged regarding HIV knowledge or consistent condom use. CONCLUSION: There was a high prevalence of sex trafficking. A history of sex trafficking was associated with a greater vulnerability to recent violence and HIV risk behaviors, underscoring the need for increased attention to the public health needs of trafficked populations. PMID- 21620403 TI - Building capacity for cervical cancer screening in outpatient HIV clinics in the Nyanza province of western Kenya. AB - OBJECTIVE: To evaluate outcomes of cervical cancer screening within HIV care and treatment clinics in Kenya. METHODS: Beginning in October 2007, visual inspection with acetic acid (VIA), colposcopy, and loop electrosurgical excision procedure (LEEP) were added to the clinical services offered at Family AIDS Care and Education Services (FACES) clinics in Kisumu, Kenya, after a systematic campaign to build capacity and community awareness. RESULTS: From October 2007 to October 2010, 3642 women underwent VIA as part of routine HIV care. Cervical intraepithelial neoplasia 2/3 was identified in 259 (7.1%) women, who were offered excisional treatment by LEEP in the clinic. Among those women offered screening, uptake was 87%. Clinical staff reported a high level of satisfaction with training for and implementation of cervical cancer screening strategies. CONCLUSION: Cervical cancer screening and prevention are feasible, acceptable, and effective within HIV care and treatment clinics. Screening test performance characteristics need to be defined for an HIV-positive population to determine the cost/benefit ratio of lower cost strategies that will ultimately be necessary to provide universal access to cervical cancer screening in low-resource settings. PMID- 21620404 TI - Mode of delivery and neonatal respiratory morbidity among HIV-exposed newborns in Latin America and the Caribbean: NISDI Perinatal-LILAC Studies. AB - OBJECTIVE: To evaluate respiratory morbidity (RM) in HIV-exposed newborns according to mode of delivery. METHODS: The NISDI Perinatal/LILAC prospective cohort studies enrolled HIV-infected pregnant women and their newborns in Latin America and the Caribbean. Associations between RM and delivery mode or other characteristics were evaluated. RESULTS: Between September 2002 and December 2009, 1630 women were enrolled, and 1443 mother-infant pairs met the inclusion criteria. There were 561 vaginal (VD), 269 cesarean before labor and membrane rupture (SCS) for preventing mother-to-child transmission (SCS-PMTCT), 248 other SCS, and 365 cesarean after labor and/or ruptured membranes (NSCS) deliveries. In total, 108 (7.5%) newborns had RM: 49 had respiratory distress syndrome (RDS), 39 had transient tachypnea (TTN), and 28 had other events (7 newborns had >1 RM event). Delivery mode was associated with RDS (P<0.005) and TTN (P<0.001). The proportion with RDS and TTN was lowest for VD (1.6% and 0.5%, respectively), highest for NSCS (4.9% and 4.7%), and intermediate for SCS-PMTCT (3.0% and 2.6%). Newborns with RDS or TTN were hospitalized longer (median +1day) than those without. A minority required ventilatory support (RDS, 24.5%-28.6%; TTN, 2.6% 15.4%). CONCLUSIONS: SCS-PMTCT is relatively safe for newborns of HIV-infected women. PMID- 21620405 TI - Potential role for organochlorine pesticides in the prevalence of peripheral arterial diseases in obese persons: results from the National Health and Nutrition Examination Survey 1999-2004. AB - OBJECTIVES: The purpose of this study was to investigate the association between organochlorine (OC) pesticides and the prevalence of peripheral arterial disease (PAD). METHODS AND RESULTS: This study included 2032 participants from the National Health and Nutrition Examination Survey (1999-2004), in which the ankle brachial index (ABI) and serum levels of OC pesticides (p,p'-DDE, trans nonachlor, oxychlordane, dieldrin, and beta-HCH) were measured simultaneously. A diagnosis of PAD was determined on the basis of an ABI<0.9. The obesity status was categorized as obese group (BMI >= 25 kg/m(2)) or non-obese group (BMI<25 kg/m(2)). Compared with subjects without PAD, those with PAD had significantly higher lipid-standardized and wet-weight concentrations of OC pesticides. Obese subjects with PAD had significantly increased mean lipid-standardization value of p,p'-DDE (OR=1.47; 95% CI, 1.08-1.99), trans-nonachlor (OR=1.68; 95% CI, 1.10 2.56), oxychlordane (OR=1.82; 95% CI, 1.09-3.03), dieldrin (OR=2.36; 95% CI, 1.69 3.31), and sums of all five pesticides (OR=1.19; 95% CI, 1.07-1.33). For non obese group, increases in the levels of OC pesticides and their sums did not account for increased risk ratio of PAD. CONCLUSION: OC pesticides are a potent risk factor for PAD. Additionally, obesity may modulate the association between OC pesticides and the development of PAD. PMID- 21620406 TI - Lipoprotein-associated phospholipase A2 testing usefulness among patients with symptomatic intracranial atherosclerotic disease. AB - BACKGROUND AND PURPOSE: Circulating lipoprotein-associated phospholipase A(2) (Lp PLA(2)) has emerged as a novel biomarker for cardiovascular diseases. Our aim was to determine Lp-PLA(2) mass and activity in a selected cohort of first-ever transient ischemic attack (TIA) or ischemic stroke patients with intracranial atherosclerotic disease (ICAD) and to investigate its relationship with the presence of classical vascular risk factors, response to secondary prevention treatments and risk of recurrent vascular events. METHODS: Lp-PLA(2) mass and activity were measured 3 months after TIA or stroke by means of the PLAC test and CAM-assay (diaDexus, Inc.) respectively in 75 patients. Classic vascular risk factors, preventive treatments and clinical characteristics at the time of the index event were recorded. Follow-up transcranial Doppler ultrasonography (TCD) was performed and the presence of a new vascular event was assessed every 6 months. RESULTS: Several preventive treatments (statins and clopidogrel) were significantly associated with lower Lp-PLA(2) mass and activity. During follow-up (median time 23 months), eighteen patients (24%) suffered a new vascular event. Baseline factors associated with new vascular events were: history of coronary artery disease, number of intracranial stenoses detected by TCD and also Lp PLA(2) activity, which was the only independent predictor for new vascular events (hazard ratio 2.89; 95% CI 1.029 to 8.096; p=0.044) after multivariate analysis (Cox regression). CONCLUSIONS: Lp-PLA(2) activity might be a useful tool to identify intracranial large-artery occlusive disease patients at higher risk of suffering new vascular events. PMID- 21620407 TI - Statistical theory of linear adsorption capillary chromatography with porous layer stationary phase. AB - A set of accurate expressions of elution-curve moments are derived from the moments of residence time and displacement in a step based on probability theory. Then the problems about residence time and displacement in a step of a solute molecule in the porous layer of capillary columns and in the moving mobile phase are described by a set of mass-balance equations respectively. The set of equations are solved in Fourier-Laplace domain, and the characteristic functions of residence time of a step, as well as the moments, are obtained by means of computing software Mathematica. At last, using numerical inverse Laplace transform, the elution curves for various conditions are calculated. In the case of large desorption constant the results entirely coincide with those of mass balance-equation theory and in the case of small desorption constant they are equivalent to those of stochastic theory. PMID- 21620408 TI - Improving quantitative gas chromatography-electron ionization mass spectrometry results using a modified ion source: demonstration for a pharmaceutical application. AB - Gas chromatography-mass spectrometry is a well established analytical technique. However, mass spectrometers with electron ionization sources may suffer from signal drifts, hereby negatively influencing quantitative performance. To demonstrate this phenomenon for a real application, a static headspace-gas chromatography method in combination with electron ionization-quadrupole mass spectrometry was optimized for the determination of residual dichloromethane in coronary stent coatings. Validating the method, the quantitative performance of an original stainless steel ion source was compared to that of a modified ion source. Ion source modification included the application of a gold coating on the repeller and exit plate. Several validation aspects such as limit of detection, limit of quantification, linearity and precision were evaluated using both ion sources. It was found that, as expected, the stainless steel ion source suffered from signal drift. As a consequence, non-linearity and high RSD values for repeated analyses were obtained. An additional experiment was performed to check whether an internal standard compound would lead to better results. It was found that the signal drift patterns of the analyte and internal standard were different, consequently leading to high RSD values for the response factor. With the modified ion source however, a more stable signal was observed resulting in acceptable linearity and precision. Moreover, it was also found that sensitivity improved compared to the stainless steel ion source. Finally, the optimized method with the modified ion source was applied to determine residual dichloromethane in the coating of coronary stents. The solvent was detected but found to be below the limit of quantification. PMID- 21620409 TI - Reduced surface area chromatography for flow-through purification of viruses and virus like particles. AB - A method for flow-through purification of viruses and virus like nano-particles using a combination of binding and size-exclusion chromatography was developed. This technique relies on minimizing the external surface area per unit volume available for virus binding by increasing the mean diameter of the beads used in the column. At the same time the impurity binding capacity of the column is maximized by utilizing beads with multiple functionalities of the optimum size. Purification of different types of viruses and virus-like-particles could be achieved using this technique. Flow-through purification of influenza virus using this technique yielded virus recoveries greater than 70-80% coupled with impurity removal greater than 80%. Finally an approach to optimize and facilitate process development using this technology is presented. Since the impurity binding occurs via a non-specific mechanism and virus recovery is achieved through reduced surface area, the technique is not limited to specific types of viruses and offers the potential as a universal purification tool. PMID- 21620410 TI - Simultaneous capillary electrophoresis competitive immunoassay for insulin, glucagon, and islet amyloid polypeptide secretion from mouse islets of Langerhans. AB - A capillary electrophoresis competitive immunoassay was developed for the simultaneous quantitation of insulin, glucagon, and islet amyloid polypeptide (IAPP) secretion from islets of Langerhans. Separation buffers and conditions were optimized for the resolution of fluorescein isothiocyanate (FITC)-labeled glucagon and IAPP immunoassay reagents, which were excited with the 488 nm line of an Ar(+) laser and detected at 520 nm with a photomultiplier tube (PMT). Cy5 labeled insulin immunoassay reagents were excited by a 635 nm laser diode module and detected at 700 nm with a separate PMT. Optimum resolution was achieved with a 20mM carbonate separation buffer at pH 9.0 using a 20 cm effective separation length with an electric field of 500 V/cm. Limits of detection for insulin, glucagon, and IAPP were 2, 3, and 3 nM, respectively. This method was used to monitor the simultaneous secretion of these peptides from as few as 14 islets after incubation in 4, 11, and 20 mM glucose for 6h. For insulin and IAPP, a statistically significant increase in secretion levels was observed, while glucagon levels were significantly reduced in the 4 and 11 mM glucose conditions. To further demonstrate the utility of the assay, the Ca(2+)-dependent secretion of these peptides was demonstrated which agreed with published reports. The ability to examine the secretion of multiple peptides may allow for the determination of regulation of secretory processes within islets of Langerhans. PMID- 21620411 TI - Study of retention behaviour and mass spectrometry compatibility in zwitterionic hydrophilic interaction chromatography for the separation of modified nucleosides and nucleobases. AB - A study has been made of the chromatographic behaviour of modified nucleosides and nucleobases using different stationary phases with functional groups of polar nature, all of them compatible with aquoorganic mobile phases. The stationary phases assayed were a pentafluorophenylpropyl (PFP) column for reverse phase separation, and another two for hydrophilic interaction chromatography (HILIC) separation. Six modified nucleosides and nucleobases (hydroxylated and methylated derivatives) were chosen as the target analytes. In the study, chromatographic resolution as well as the sensitivity in detection by mass spectrometry were taken into account. The results obtained showed that the zwitterionic (ZIC-HILIC) column was the most suitable one for the separation of these analytes. From the study of the different parameters affecting separation it may be concluded that in the ZIC-HILIC column separation is based on a mechanism of partition and interaction through weak electrostatic forces. PMID- 21620412 TI - Kinetics and thermodynamic study of aniline adsorption by multi-walled carbon nanotubes from aqueous solution. AB - Multi-walled carbon nanotubes (MWCNTs) were used in the adsorptive removal of aniline, an organic pollutant, from an aqueous solution. It was found that carbon nanotubes with a higher specific surface area adsorbed and removed more aniline from an aqueous solution. The adsorption was dependent on factors, such as MWCNTs dosage, contact time, aniline concentration, solution pH and temperature. The adsorption study was analyzed kinetically, and the results revealed that the adsorption followed pseudo-second order kinetics with good correlation coefficients. In addition, it was found that the adsorption of aniline occurred in two consecutive steps, including the slow intra-particle diffusion of aniline molecules through the nanotubes. Various thermodynamic parameters, including the Gibbs free energy change (DeltaG degrees ), enthalpy change (DeltaH degrees ) and entropy change (DeltaS degrees ), were calculated. The results indicated that the spontaneity of the adsorption, exothermic nature of the adsorption and the decrease in the randomness reported as DeltaG degrees , DeltaH degrees and DeltaS degrees , respectively, were all negative. PMID- 21620414 TI - Measuring and inducing brain plasticity in chronic aphasia. AB - Brain plasticity associated with anomia recovery in aphasia is poorly understood. Here, I review four recent studies from my lab that focused on brain modulation associated with long-term anomia outcome, its behavioral treatment, and the use of transcranial brain stimulation to enhance anomia treatment success in individuals with chronic aphasia caused by left hemisphere stroke. In a study that included 15 participants with aphasia who were compared to a group of 10 normal control subjects, we found that improved naming ability was associated with increased left hemisphere activity. A separate study (N = 26) revealed similar results in that improved anomia treatment outcome was associated with increased left hemisphere recruitment. Taken together, these two studies suggest that improved naming in chronic aphasia relies on the damaged left hemisphere. Based on these findings, we conducted two studies to appreciate the effect of using low current transcranial electrical stimulation as an adjuvant to behavioral anomia treatment. Both studies yielded positive findings in that anomia treatment outcome was improved when it was coupled with real brain stimulation as compared with a placebo (sham) condition. Overall, these four studies support the notion that the intact cortex in the lesioned left hemisphere supports anomia recovery in aphasia. LEARNING OUTCOMES: Readers will (a) be able to appreciate the possible influence of animal research upon the understanding of brain plasticity induced by aphasia treatment, (b) understand where functional changes associated with anomia treatment occur in the brain, (c) understand the basic principles of transcranial direct current stimulation, and (d) understand how brain stimulation coupled with aphasia treatment may potentially improve treatment outcome. PMID- 21620413 TI - Experience-dependent neural plasticity in the adult damaged brain. AB - Behavioral experience is at work modifying the structure and function of the brain throughout the lifespan, but it has a particularly dramatic influence after brain injury. This review summarizes recent findings on the role of experience in reorganizing the adult damaged brain, with a focus on findings from rodent stroke models of chronic upper extremity (hand and arm) impairments. A prolonged and widespread process of repair and reorganization of surviving neural circuits is instigated by injury to the adult brain. When experience impacts these same neural circuits, it interacts with degenerative and regenerative cascades to shape neural reorganization and functional outcome. This is evident in the cortical plasticity resulting from compensatory reliance on the "good" forelimb in rats with unilateral sensorimotor cortical infarcts. Behavioral interventions (e.g., rehabilitative training) can drive functionally beneficial neural reorganization in the injured hemisphere. However, experience can have both behaviorally beneficial and detrimental effects. The interactions between experience-dependent and injury-induced neural plasticity are complex, time dependent, and varied with age and other factors. A better understanding of these interactions is needed to understand how to optimize brain remodeling and functional outcome. LEARNING OUTCOMES: Readers will be able to describe (a) experience effects that are maladaptive for behavioral outcome after brain damage, (b) manipulations of experience that drive functionally beneficial neural plasticity, and (c) reasons why rehabilitative training effects can be expected to vary with age, training duration and timing. PMID- 21620415 TI - Children's human figure drawings do not measure intellectual ability. AB - Children typically follow a well-defined series of stages as they learn to draw, but the rate at which they progress through these stages varies from child to child. Some experts have argued that these individual differences in drawing development reflect individual differences in intelligence. Here we assessed the validity of a drawing test that is commonly used to assess children's intellectual abilities. In a single study, 125 5- and 6-year-olds completed the Draw-A-Person: A Quantitative Scoring System (DAP:QSS) and the Wechsler Preschool and Primary Scale of Intelligence-Revised (WPPSI-R) or the Wechsler Abbreviated Scale of Intelligence (WASI). Although there was a statistically significant correlation between scores on the DAP:QSS and scores on the Wechsler tests, when the scores of individual children were examined, the DAP:QSS yielded a high number of false positives and false negatives for low intellectual functioning. We conclude that the DAP:QSS is not a valid measure of intellectual ability and should not be used as a screening tool. PMID- 21620416 TI - Efficacy of vitamin D3 as add-on therapy in patients with relapsing-remitting multiple sclerosis receiving subcutaneous interferon beta-1a: a Phase II, multicenter, double-blind, randomized, placebo-controlled trial. AB - Recent studies have demonstrated the immunomodulatory properties of vitamin D, and vitamin D deficiency may be a risk factor for the development of MS. The risk of developing MS has, in fact, been associated with rising latitudes, past exposure to sun and serum vitamin D status. Serum 25-hydroxyvitamin D [25(OH)D] levels have also been associated with relapses and disability progression. The identification of risk factors, such as vitamin D deficiency, in MS may provide an opportunity to improve current treatment strategies, through combination therapy with established MS treatments. Accordingly, vitamin D may play a role in MS therapy. Small clinical studies of vitamin D supplementation in patients with MS have reported positive immunomodulatory effects, reduced relapse rates and a reduction in the number of gadolinium-enhancing lesions. However, large randomized clinical trials of vitamin D supplementation in patients with MS are lacking. SOLAR (Supplementation of VigantOL((r)) oil versus placebo as Add-on in patients with relapsing-remitting multiple sclerosis receiving Rebif((r)) treatment) is a 96-week, three-arm, multicenter, double-blind, randomized, placebo-controlled, Phase II trial (NCT01285401). SOLAR will evaluate the efficacy of vitamin D(3) as add-on therapy to subcutaneous interferon beta-1a in patients with RRMS. Recruitment began in February 2011 and is aimed to take place over 1 calendar year due to the potential influence of seasonal differences in 25(OH)D levels. PMID- 21620417 TI - Intrathoracic phrenic pacing: a 10-year experience in France. AB - BACKGROUND: Phrenic pacing is an alternative to positive-pressure ventilation in selected patients, mostly in cases of upper spinal cord injury. We evaluated results of phrenic pacing performed by video-assisted thoracic surgery (VATS). METHOD: Between 1997 and 2007, after complete neuromuscular investigations, 20 patients requiring full-time ventilation were selected for phrenic pacing (19 with posttraumatic tetraplegia and 1 with congenital central hypoventilation syndrome). Quadripolar cuff electrodes were fixed around each intrathoracic phrenic nerve via bilateral VATS. They were connected to a subcutaneous radiofrequency receiver coupled to an external radiofrequency transmitter. All patients participated in a reconditioning program beginning 2 weeks after implantation and continued until ventilatory weaning. RESULTS: Phrenic pacing was successful in all cases. No intraoperative complications or perioperative mortality were observed. Intraoperative testing detected stimulation thresholds in 19 patients (range, 0.05-2.9 mA). Ventilatory weaning was obtained in 18 patients. Median diaphragm reconditioning time was 6 weeks (2 weeks-11 months). Reconditioning was still in process in a young woman and was not achieved in an elderly woman with a 4-year history of tetraplegia. All the patients weaned from mechanical ventilation reported improved quality of life. Failure or delay in recovery of effective diaphragm contraction was due to nonreversible amyotrophy. CONCLUSIONS: VATS implantation of 4-pole electrodes around the intrathoracic phrenic nerve is a safe procedure. Ventilatory weaning correlates with the degree of diaphragmatic amyotrophy. Phrenic pacing, performed as soon as neurologic and orthopedic stabilization is achieved, is the most important prognostic factor for successful weaning. PMID- 21620418 TI - Detection of integrin-linked kinase in the serum of patients with malignant pleural mesothelioma. AB - OBJECTIVE: Integrin-linked kinase, which is relevant to neoplastic transformation, is highly expressed in malignant pleural mesothelioma. Recently, detection of integrin-linked kinase in serum of patients with ovarian cancer has been reported. This study asks whether integrin-linked kinase can also be detected in serum of patients with malignant pleural mesothelioma and whether serum level has diagnostic or prognostic relevance for that disease. METHODS: A sandwich enzyme-linked immunosorbent assay was designed to detect integrin-linked kinase and applied to serum samples from 46 patients with malignant pleural mesothelioma, 98 patients with other malignant chest disease, and 23 patients with benign chest disease. Integrin-linked kinase serum concentration and clinical data were correlated statistically. RESULTS: Median serum integrin linked kinase concentration was significantly higher in malignant pleural mesothelioma (8.89 ng/mL) than in other malignant chest disease (0.66 ng/mL) or benign chest disease (0.78 ng/mL, P < .001). There was no relevant correlation of serum integrin-linked kinase with cell lysis parameters (R(2) < 0.1). Serum integrin-linked kinase concentration greater than 2.48 ng/mL had diagnostic sensitivity of 80%, specificity of 95%, positive predictive value of 85.7%, negative predictive value of 92.7%, and overall accuracy of 91% for distinction between malignant pleural mesothelioma and other diseases. Serum integrin-linked kinase concentration in malignant pleural mesothelioma was independent of histologic subtype or asbestos exposure. There was no statistically significant impact of serum integrin-linked kinase concentration on prognosis. CONCLUSIONS: Integrin-linked kinase can be detected in serum of patients with malignant pleural mesothelioma and may be a diagnostic marker for the disease. PMID- 21620419 TI - Quantity, distribution, and impacts of coastal driftwood triggered by a typhoon. AB - Typhoon Morakot pounded Taiwan in 2009 with record-breaking rainfall, washing an unprecedented amount of driftwood into the sea that was partially deposited at the coastal areas. According to the satellite imagery analysis, more than three million trees fell and were washed away to occupy 83.2% of the Taiwanese coastline, including 52 fishing harbors. The amount cleaned-up was only 1/7 of the total coastal driftwood. It was found that the amount of coastal driftwood is not only related to the amount of precipitation but is also related to the distance from the location of the landslide to the river mouth and to the landslide area. The amount of accumulated coastal driftwood demonstrated log profile declines with increasing distance to the river mouth. Nearshore current and wave motion are the critical factors for driftwood deposition. Much of the driftwood washed into the sea harmed the tourism and fishing industries, endangered navigation and oceanic activities, and impacted the marine environment and ecosystem. PMID- 21620420 TI - Microbial community dynamics and biodegradation of polycyclic aromatic hydrocarbons in polluted marine sediments in Hong Kong. AB - Dynamics of microbial community and biodegradation of polycyclic aromatic hydrocarbons (PAHs) in polluted marine sediments, artificially spiked with a mixture of PAHs (fluorene, phenanthrene, fluoranthene and pyrene), were examined for a period of 60 days. Microbial communities were characterised by bacterial counts, ester-linked fatty acid methyl ester (EL-FAME) analysis and denaturing gradient gel electrophoresis (DGGE). A noted reduction in species diversity occurred only in the high PAH level treatment at onset. Both EL-FAME and DGGE demonstrated a marked shift in microbial community, in all the PAH level treatments, afterwards, with increases in the number of fatty acid degraders, the relative abundance of fatty acid biomarkers for gram-negative bacteria and a decrease in species diversity. The shift was also accompanied by the significant decrease in PAH concentrations. By the end of the experiment, diversity indices, based on both approaches, recovered when PAH concentrations declined to their background levels, except in the high PAH level treatment. PMID- 21620421 TI - Effects of nutrient enrichment on macroalgal coralligenous assemblages. AB - Effects of eutrophication on marine ecosystems have been widely studied, even if both the effects on deep subtidal rocky assemblages and response of different successional stages to nutrients impact are still not clear. In this context, the study aimed to evaluate the effects of nutrient enrichment on Mediterranean macroalgal assemblages associated with coralligenous habitat. A manipulative field experiment was carried out by supplying both mature and early successional stages of assemblages with nutrients. A total of 62 macroalgal species were identified. Multivariate and univariate analyses showed that the structure of both mature and early successional macroalgal assemblages of coralligenous significantly varied between areas treated with nutrients and not treated areas. Moreover, differences were stronger when macroalgal assemblages were in the early successional stage than in the mature one. Results highlighted the role played by nutrients in determining the structure of macroalgal coralligenous assemblages, furthermore suggesting possible synergetic effects with other kinds of disturbances. PMID- 21620422 TI - Mussel farming as a nutrient reduction measure in the Baltic Sea: consideration of nutrient biogeochemical cycles. AB - Nutrient loads from the land to the sea must be reduced to combat coastal eutrophication. It has been suggested that further mitigation efforts are needed in the brackish Baltic Sea to decrease nutrients, especially in eutrophic coastal areas. Mussel farming is a potential measure to remove nutrients directly from the sea. Mussels consume phytoplankton containing nitrogen (N) and phosphorus (P); when the mussels are harvested these nutrients are removed from the aquatic system. However, sedimentation of organic material in faeces and pseudo-faeces below a mussel farm consumes oxygen and can lead to hypoxic or even anoxic sediments causing an increased sediment release of ammonium and phosphate. Moreover, N losses from denitrification can be reduced due to low oxygen and reduced numbers of bioturbating organisms. To reveal if mussel farming is a cost effective mitigation measure in the Baltic Sea the potential for enhanced sediment nutrient release must be assessed. PMID- 21620423 TI - The use of Lagrangian trajectories for the identification of the environmentally safe fairways. AB - We propose and test a method for the optimisation of marine fairways to minimise the risk to high-value areas, based on statistical analysis of Lagrangian trajectories of current-driven pollution transport. The offshore areas are quantified according to the probability of pollution released in these areas to reach vulnerable regions. The method contains an eddy-resolving circulation model, a scheme for tracking of Lagrangian trajectories, a technique for the calculation of quantities characterising the potential of different sea areas to supply adverse impacts, and routines to construct the optimum fairway. The gain is expressed in terms of the probability of pollution transport to the nearshore and the associated time (particle age). The use of the optimum fairway would decrease the probability of coastal pollution by 40% or increase the average time of reaching the pollution to the coast from 5.3 to about 9 days in the Gulf of Finland, the Baltic Sea. PMID- 21620424 TI - Biosynthesis and localization of parthenolide in glandular trichomes of feverfew (Tanacetum parthenium L. Schulz Bip.). AB - Feverfew (Tanacetum parthenium) is a perennial medicinal herb and is a rich source of sesquiterpene lactones. Parthenolide is the main sesquiterpene lactone in feverfew and has attracted attention because of its medicinal potential for treatment of migraine and cancer. In the present work the parthenolide content in different tissues and developmental stages of feverfew was analyzed to study the timing and localization of parthenolide biosynthesis. The strongest accumulating tissue was subsequently used to isolate sesquiterpene synthases with the goal to isolate the gene encoding the first dedicated step in parthenolide biosynthesis. This led to the isolation and charachterization of a germacrene A synthase (TpGAS) and an (E)-beta-caryophyllene synthase (TpCarS). Transcript level patterns of both sesquiterpene synthases were analyzed in different tissues and glandular trichomes. Although TpGAS was expressed in all aerial tissues, the highest expression was observed in tissues that contain high concentrations of parthenolide and in flowers the highest expression was observed in the biosynthetically most active stages of flower development. The high expression of TpGAS in glandular trichomes which also contain the highest concentration of parthenolide, suggests that glandular trichomes are the secretory tissues where parthenolide biosynthesis and accumulation occur. PMID- 21620425 TI - Coping with toxic plant compounds--the insect's perspective on iridoid glycosides and cardenolides. AB - Specializing on host plants with toxic secondary compounds enforces specific adaptation in insect herbivores. In this review, we focus on two compound classes, iridoid glycosides and cardenolides, which can be found in the food plants of a large number of insect species that display various degrees of adaptation to them. These secondary compounds have very different modes of action: Iridoid glycosides are usually activated in the gut of the herbivores by beta-glucosidases that may either stem from the food plant or be present in the gut as standard digestive enzymes. Upon cleaving, the unstable aglycone is released that unspecifically acts by crosslinking proteins and inhibiting enzymes. Cardenolides, on the other hand, are highly specific inhibitors of an essential ion carrier, the sodium pump. In insects exposed to both kinds of toxins, carriers either enabling the safe storage of the compounds away from the activating enzymes or excluding the toxins from sensitive tissues, play an important role that deserves further analysis. To avoid toxicity of iridoid glycosides, repression of activating enzymes emerges as a possible alternative strategy. Cardenolides, on the other hand, may lose their toxicity if their target site is modified and this strategy has evolved multiple times independently in cardenolide-adapted insects. PMID- 21620426 TI - Homology modeling of the three membrane proteins of the dhurrin metabolon: catalytic sites, membrane surface association and protein-protein interactions. AB - Formation of metabolons (macromolecular enzyme complexes) facilitates the channelling of substrates in biosynthetic pathways. Metabolon formation is a dynamic process in which transient structures mediated by weak protein-protein interactions are formed. In Sorghum, the cyanogenic glucoside dhurrin is derived from l-tyrosine in a pathway involving the two cytochromes P450 (CYPs) CYP79A1 and CYP71E1, a glucosyltransferase (UGT85B1), and the redox partner NADPH dependent cytochrome P450 reductase (CPR). Experimental evidence suggests that the enzymes of this pathway form a metabolon. Homology modeling of the three membrane bound proteins was carried out using the Sybyl software and available relevant crystal structures. Residues involved in tight positioning of the substrates and intermediates in the active sites of CYP79A1 and CYP71E1 were identified. In both CYPs, hydrophobic surface domains close to the N-terminal trans-membrane anchor and between the F' and G helices were identified as involved in membrane anchoring. The proximal surface of both CYPs showed positively charged patches complementary to a negatively charged bulge on CPR carrying the FMN domain. A patch of surface exposed, positively charged amino acid residues positioned on the opposite face of the membrane anchor was identified in CYP71E1 and might be involved in binding UGT85B1 via a hypervariable negatively charged loop in this protein. PMID- 21620427 TI - Potential public health significance of faecal contamination and multidrug resistant Escherichia coli and Salmonella serotypes in a lake in India. AB - OBJECTIVE: To assess the prevalence of faecal coliform bacteria and multiple drug resistance among Escherichia coli and Salmonella serotypes from Vembanadu Lake. STUDY DESIGN: Systematic microbiological testing. METHODS: Monthly collection of water samples were made from ten stations on the southern and northern parts of a salt water regulator constructed in Vembanadu Lake in order to prevent incursion of seawater during certain periods of the year. Density of faecal colifrom bacteria was estimated. E. coli and Salmonella were isolated and their different serotypes were identified. Antibiotic resistance analysis of E. coli and Salmonella serotypes was done and the MAR index of individual isolates was calculated. RESULTS: Density of faecal coliform bacteria ranged from mean MPN value 2900 -7100/100ml. Results showed multiple drug resistance pattern among the bacterial isolates. E. coli showed more than 50% resistance to amickacin, oxytetracycline, streptomycin, tetracycline and kanamycin while Salmonella showed high resistance to oxytetracycline, streptomycin, tetracycline and ampicillin. The MAR indexing of the isolates showed that they have originated from high risk source such as humans, poultry and dairy cows. CONCLUSIONS: The high density of faecal coliform bacteria and prevalence of multi drug resistant E. coli and Salmonella serotypes in the lake may pose severe public health risk through related water borne and food borne outbreaks. PMID- 21620428 TI - Impact of probiotic administration on the health and fecal microbiota of young calves: a meta-analysis of randomized controlled trials of lactic acid bacteria. AB - Before weaning, dairy calves are susceptible to many pathogens which can affect their subsequent performance. The use of lactic acid bacteria (LAB) has been identified as a tool to maintain the intestinal microbial balance and to prevent the establishment of opportunistic pathogenic bacterial populations. However, a consensus has not been reached as to whether probiotics may be effective in reducing the prevalence of gastrointestinal diseases in young calves. The aim of this meta-analysis was to assess the effect of probiotics on diarrhea incidence and the intestinal microbial balance. LAB supplementation has been shown to exert a protective effect and to reduce the incidence of diarrhea (relative risk, RR=0.437, 95% confidence interval (CI) 0.251-0.761). In the subanalysis, this protective effect of the probiotics against diarrhea was observed only in trials that used whole milk (RR=0.154, 95% CI 0.079-0.301) and trials that used multistrain inocula (RR=0.415, 95% CI 0.227-0.759). Probiotics did not improve the fecal characteristics (standardized mean difference, SMD=-0.4904, 95% CI 1.011-0.035) and were unable to change the LAB:coliforms ratio (SMD=0.016, 95% CI -0.701-0.733). Probiotics showed a beneficial impact on the LAB:coliforms ratio in the subanalysis that included trials that used whole milk (SMD=0.780, 95% CI 0.141-1.418) and monostrain inocula (SMD=0.990, 95% CI 0.340-1.641). The probability of significant effects (probiotic positive effect) in a new study was >0.70 for diarrhea and fecal consistency. Whole milk feeding improved the action of the probiotic effect on the incidence of diarrhea and LAB:coliforms ratio. The probability to find significant effects in the diarrhea frequency and LAB:coliforms ratio was higher (P>0.85) if the new studies were conducted using whole milk to feed calves. This paper defines the guidelines to standardize the experimental designs of future trials. LAB can be used as growth promoters in calves instead of antibiotics to counteract the negative effects of their widespread use. PMID- 21620429 TI - Primary and secondary lacrimal canaliculitis: a review of literature. AB - Canaliculitis is an uncommon inflammation of the proximal lacrimal drainage system that is frequently misdiagnosed. It classically presents with symptoms of unilateral conjunctivitis, mucopurulent discharge, medial canthal inflammation, epiphora, and a red, pouting punctum. We summarize the literature on canaliculitis published from antiquity to the modern era and explore therapeutic options. PMID- 21620430 TI - Pregnancy and glaucoma. AB - Glaucoma, primarily a disease of the older population, may affect women of childbearing age. Pregnancy affects the intraocular pressure (IOP) of women with pre-existing glaucoma. Both elevations and reductions of IOP have been reported during pregnancy. Additionally, visual field test results may fluctuate during pregnancy. In managing the pregnant glaucoma patient with medical therapy, one must consider not only the systemic side effects on the mother, but also any potentially harmful effects on the developing fetus. All anti-glaucoma medications are categorized as class C by the Food and Drug Administration, except brimonidine and nonspecific adrenergic agonists, which belong to class B. In general, the lowest effective dosage of medication should be used. Systemic absorption can be reduced by punctal occlusion, eyelid closure, and blotting the excess drops away during administration. In those patients who need surgery, most local anesthetics may be used safely because they have not been shown to be teratogenic in humans. Antifibrotic agents commonly used adjunctively in trabeculectomy, however, should be avoided. Glaucoma laser procedures, such as laser peripheral iridotomy and laser trabeculoplasty, have been employed without identifiable teratogenic effects or increased risk of side effects for pregnant women. PMID- 21620431 TI - HIV-protease inhibitors block the replication of both vesicular stomatitis and influenza viruses at an early post-entry replication step. AB - The inhibitors of HIV-1 protease (PIs) have been designed to block the activity of the viral aspartyl-protease. However, it is now accepted that this family of inhibitors can also affect the activity of cell proteases. Since the replication of many virus species requires the activity of host cell proteases, investigating the effects of PIs on the life cycle of viruses other than HIV would be of interest. Here, the potent inhibition induced by saquinavir and nelfinavir on the replication of both vesicular stomatitis and influenza viruses is described. These are unrelated enveloped RNA viruses infecting target cells upon endocytosis and intracellular fusion. The PI-induced inhibition was apparently a consequence of a block at the level of the fusion between viral envelope and endosomal membranes. These findings would open the way towards the therapeutic use of PIs against enveloped RNA viruses other than HIV. PMID- 21620432 TI - Efficient electricity production and simultaneously wastewater treatment via a high-performance photocatalytic fuel cell. AB - A great quantity of wastewater were discharged into water body, causing serious environmental pollution. Meanwhile, the organic compounds in wastewater are important sources of energy. In this work, a high-performance short TiO(2) nanotube array (STNA) electrode was applied as photoanode material in a novel photocatalytic fuel cell (PFC) system for electricity production and simultaneously wastewater treatment. The results of current work demonstrate that various model compounds as well as real wastewater samples can be used as substrates for the PFC system. As a representative of model compounds, the acetic acid solution produces the highest cell performance with short-circuit current density 1.42 mA cm(-2), open-circuit voltage 1.48 V and maximum power density output 0.67 mW cm(-2). The STNA photoanode reveals obviously enhanced cell performance compared with TiO(2) nanoparticulate film electrode or other long nanotubes electrode. Moreover, the photoanode material, electrolyte concentration, pH of the initial solution, and cathode material were found to be important factors influencing the system performance of PFC. Therefore, the proposed fuel cell system provides a novel way of energy conversion and effective disposal mode of organics and serves well as a promising technology for wastewater treatment. PMID- 21620433 TI - Antibiotics pollution in Jiulong River estuary: source, distribution and bacterial resistance. AB - To gain insight into the antibiotic pollution in the Jiulong River estuary and the pollutant sources, we analyzed the concentration of 22 widely-used antibiotics in water samples collected from the river and estuary, 17 and 18 sampling sites, respectively. Contamination with sulfonamides, quinolones and chloramphenicols was frequently detected and the distribution pattern of antibiotics suggested that most of the pollutants are from the Jiulong River, especially from the downstream watersheds. To reveal the ecological effects, we isolated 35 bacterial strains from the estuary and analyzed their antibiotic resistance to the eight most frequently detected antibiotics. The bacteria were subsequently classified into seven different genera by 16SrDNA sequencing. Up to 97.1% of the bacteria showed resistance and 70.6% of strains showed multi resistance to these antibiotics, especially to sulfonamides. This study demonstrated a pattern of antibiotic contamination in the Jiulong River and its estuary and illustrated high bacterial antibiotic resistance which was significantly correlated with the average antibiotics concentrations and detected frequencies in the estuary. PMID- 21620434 TI - Influences of influent carbon source on extracellular polymeric substances (EPS) and physicochemical properties of activated sludge. AB - It is necessary to understand the bioflocculation, settling and dewatering characteristics in the activated sludge process in order to establish more efficient operational strategies. The influences of carbon source on the extracellular polymeric substances (EPS) and flocculation, settling and dewatering properties of the activated sludge were investigated. Laboratory-scale completely mixed activated sludge processes were used to grow the activated sludge with different carbon sources of starch, glucose and sodium acetate. The sludge fed with acetate had highest loosely bound EPS (LB-EPS) and that fed with starch lowest. The amount of tightly bound EPS (TB-EPS), protein content in LB EPS, polysaccharide content and protein contents in TB-EPS, were independent of the influent carbon source. The polysaccharide content in LB-EPS of the activated sludge fed with sodium acetate was lower slightly than those of starch and glucose. The sludge also had a nearly consistent flocs size and the sludge volume index (SVI) value. ESS content of the sludge fed with sodium acetate was higher initially, although it was similar to those fed with glucose and starch finally. However, the specific resistance to filtration and normalized capillary suction time fluctuated first, but finally were stable at around 5.0*10(8)mkg(-1) and 3.5 s Lg(-1) SS, respectively. Only the protein content in LB-EPS weakly correlated with the flocs size and SVI of the activated sludge. But there was no correlation between any other EPS contents or components and the physicochemical properties of the activated sludge. PMID- 21620435 TI - Mercury (Hg) emissions from domestic biomass combustion for space heating. AB - Three mercury (Hg) species (gaseous elemental mercury (GEM), gaseous oxidized mercury (GOM), and fine particulate-bound mercury (PBM(2.5))) were measured in the stack of a small scale wood combustion chamber at 400 degrees C, in the stack of an advanced wood boiler, and in two areas influenced by wood combustion. The low temperature process (lab-scale) emitted mostly GEM (~99% when burning wood pellets and ~95% when burning unprocessed wood). The high temperature wood boiler emitted a greater proportion of oxidized Hg (approximately 65%) than the low temperature system. In field measurements, mean PBM(2.5) concentrations at the rural and urban sites in winter were statistically significantly higher than in warmer seasons and were well correlated with Delta-C concentrations, a wood combustion indictor measured by an aethalometer (UV-absorbable carbon minus black carbon). Overall the results suggest that wood combustion may be an important source of oxidized mercury (mostly in the particulate phase) in northern climates in winter. PMID- 21620436 TI - Stoichiometric and molecular evidence for the enrichment of anaerobic ammonium oxidizing bacteria from wastewater treatment plant sludge samples. AB - Anammox enrichments were readily developed from seven municipal wastewater treatment plants (WWTPs) sludge, but not with methanogenic granular sludge from two agro-industrial WWTPs. Only 50d was required for the first evidence of anammox activity from a return activated sludge obtained from a WWTP operated for nutrient removal. The molar ratios of nitrite and ammonium consumption of approximately 1.32 as well as nitrate and dinitrogen gas product ratios of approximately 0.095 provided evidence of the anammox reaction. The presence of anammox was confirmed by polymerase chain reaction (PCR) using primer sets (PLA46F and AMX820R) specific for anammox bacteria. The 16S rRNA gene fragment of anammox bacteria was detected in seven enrichment cultures (ECs) with demonstrated anammox activity but not in the original inocula from which the ECs were derived and also not in the two methanogenic sludge samples, which indicates the PCR predicted the anammox activity. Two genera, Brocadia and Kuenenia, were successfully identified as the Planctomycetes occurring in the clone libraries of successful anammox enrichments. Brocadia dominated in cultures that were respiked extensively; whereas Kuenenia predominated in cultures that were less aggressively respiked. These findings indicate that respiking management may play an important role on selecting the genus of anammox bacteria. The batch enrichment results clearly illustrate that anammox can be readily enriched from municipal sludge from a wide variety of process operations at WWTPs. PMID- 21620437 TI - Genomic data reject the hypothesis of a prosimian primate clade. AB - The phylogenetic position of tarsiers within the primates has been a controversial subject for over a century. Despite numerous morphological and molecular studies, there has been weak support for grouping tarsiers with either strepsirrhine primates in a prosimian clade or with anthropoids in a haplorrhine clade. Here, we take advantage of the recently released whole genome assembly of the Philippine tarsier, Tarsius syrichta, in order to infer the phylogenetic relationship of Tarsius within the order Primates. We also present estimates of divergence times within the primates. Using a 1.26 million base pair multiple sequence alignment derived from 1078 orthologous genes, we provide overwhelming statistical support for the presence of a haplorrhine clade. We also present divergence date estimates using local relaxed molecular clock methods. The estimated time of the most recent common ancestor of extant Primates ranged from 64.9 Ma to 72.6 Ma, and haplorrhines were estimated to have a most recent common ancestor between 58.9 Ma and 68.6 Ma. Examination of rates of nucleotide substitution in the three major extant primate clades show that anthropoids have a slower substitution rate than either strepsirrhines or tarsiers. Our results provide the framework on which primate morphological, reproductive, and genomic features can be reconstructed in the broader context of mammalian phylogeny. PMID- 21620438 TI - The role of IL-6, IL-8 and MCP-1 and their promoter polymorphisms IL-6 -174GC, IL 8 -251AT and MCP-1 -2518AG in the risk of venous thromboembolism: a case-control study. AB - INTRODUCTION: Cytokines increased the risk of venous thromboembolism (VTE) in some case-control studies, but not in a prospective study. Data concerning the role of cytokines in the risk of VTE are limited. We examined in a case-control study the association of VTE and levels of interleukin (IL)-6, IL-8 and monocyte chemotactic protein-1 (MCP-1) and assessed whether promoter polymorphisms (IL-6 174GC, IL-8 -251AT, MCP-1 -2518AG) would affect the thrombotic risk and cytokine levels. MATERIALS AND METHODS: The study included 119 patients (94 women) with a first event of VTE aged between 18-60 years, and 126 healthy controls (100 women) matched for age (+/-5 years). Blood was collected >7 months after the thrombotic event. Odds ratios (ORs) were calculated per increase of cytokines levels by 1 pg/mL. RESULTS: ORs adjusted for age and sex were 1.520 [95% Confidence Interval (CI) 1.177 - 1.962] for IL-6, 1.095 (95% CI 1.002 - 1.196) for IL-8 and 1.000 (0.988 - 1.012) for MCP-1. With additional adjustment for ethnic composition, body mass index (BMI) and high sensitive C-reactive protein (hs-CRP), risk estimates remained significant for IL-6 and became of borderline statistical significance for IL-8. Polymorphisms did not influence the thrombotic risk and the cytokine levels in study participants. CONCLUSION: VTE was associated with IL 6 and IL-8 levels, and for IL-6 this association was independent of BMI and hs CRP. Thus far, a causal relationship between inflammation and VTE remains to be clarified and more prospective data are warranted. PMID- 21620439 TI - Combined use of the high heparin step and optical density to optimize diagnostic sensitivity and specificity of an anti-PF4/heparin enzyme-immunoassay. AB - BACKGROUND: IgG-specific anti-PF4/heparin enzyme-immunoassays (EIAs) are sensitive but not specific for platelet-activating antibodies, the cause of heparin-induced thrombocytopenia (HIT). Two features of EIA reactivity predict for presence of HIT antibodies - the magnitude of a positive result (in optical density [OD] units) and the inhibition of reactivity at high heparin concentrations - but their combined utility remains uncertain. OBJECTIVE: To determine for an IgG-specific EIA how the OD values of a positive reaction and its inhibition by high heparin can be optimally combined. METHODS: We screened 1,000 consecutive patients with suspected HIT using an IgG-specific PF4/heparin in-house EIA with and without high heparin (100 IU/mL); and by the heparin induced platelet activation test. RESULTS: Platelet-activating antibodies were rarely detected (<0.2%) when the IgG-specific EIA was negative at the conventional cut-off (OD, 0.5). However, an OD cut-off of 1.0 resulted in an unacceptable loss of sensitivity (14/83=17%) for detecting platelet-activating antibodies. The high heparin step increased specificity for platelet-activating antibodies from 72% to 89% without loss of sensitivity when applied to weak positive sera (OD<=1.0). However, decreased sensitivity was observed with strong positive sera (OD>1.0): 11/69 such sera (16%) that did not show >40% inhibition by high heparin nevertheless contained platelet-activating antibodies. CONCLUSION: Specificity of an IgG-specific EIA for detecting platelet-activating antibodies can be optimized by applying the high heparin inhibition step to weak positive reactions (0.5-<=1.0 OD). However, applying the high heparin inhibition step to strong-positive reactions (>1.0 OD) in our in-house assay risks falsely classifying a serum as negative for platelet-activating antibodies. PMID- 21620440 TI - Normal range of mean platelet volume in healthy subjects: Insight from a large epidemiologic study. AB - AIM: Mean platelet volume (MPV) in the healthy population has not been studied before. Therefore, the aim of the study was to measure MPV in normal subjects in a large cohort of Turkish adults. METHODS: A total of 2298 subjects with a mean age of 50 (age range 18 to 92) were interviewed. Subjects who had smoking habit, diabetes, hypertension, coronary artery disease, dyslipidemia, chronic obstructive pulmonary disease, cancer, chronic use of any drugs including antiplatelets, heavy drinkers, metabolic syndrome, ejection fraction <55%, creatinine >1.4 in men and >1.1 in women, abnormal liver function tests and an abnormal TSH were excluded in a in a stepwise manner. Complete blood counts were done on the same day within 6 hours by a CELL-DYN 3700 SL analyzer (Abbott Diagnostics). RESULTS: Three hundred twenty-six participants (204 females (63%) and 122 males (37%) with a mean age of 41 +/- 16) constituted the final healthy cohort. Mean MPV of the cohort was 8.9 +/- 1.4 fL. There was no significant difference among age groups regarding MPV. CONCLUSION: Ninety-five percent of the individuals had a MPV between 7.2 and 11.7 fL. A patient having a MPV beyond this range should be evaluated carefully especially for occlusive arterial diseases. PMID- 21620441 TI - Glomerular filtration rate in patients with atrial fibrillation on warfarin treatment: a subgroup analysis from the AURICULA registry in Sweden. AB - INTRODUCTION: Numerous associations between chronic kidney disease (CKD) and atrial fibrillation (AF) have been reported and patients with CKD on anticoagulation therapy have an increased risk of bleeding. Currently, new anticoagulant agents are emerging in clinical practice, some of which are excreted by the kidneys. The proportion of AF patients on anticoagulant treatment with reduced renal function is, however, unknown. MATERIALS AND METHODS: Using AURICULA, a Swedish registry for anticoagulation, estimated glomerular filtration rate (eGFR) was investigated in AF patients on warfarin treatment (n = 2,603). The study group was compared with a healthy sample from the population (n = 2,261). Two different creatinine prediction equations were used for calculating eGFR: the Lund-Malmo (LM) and MDRD Study equation. RESULTS: The fraction of AF patients with eGFR <30 and <45 ml/min/1.73 m(2) were 8.1% and 22.9% with the LM and 4.3% and 16.3% with the MDRD equation, respectively, and significantly higher than corresponding values in the reference population. GFR decreased with increasing age, where 11.4% and 5.6% of AF patients aged >= 75 years had eGFR <30 ml/min/1.73 m(2) according to the LM and MDRD equations, respectively. CONCLUSIONS: Severe renal impairment is common among AF patients on anticoagulant treatment with warfarin, especially at higher ages. Monitoring of renal function should be implemented in clinical practice for AF patients treated with new anticoagulants eliminated by the kidneys. PMID- 21620442 TI - Safety and efficacy of warfarin in paediatric patients with prosthetic cardiac valves: a retrospective audit. AB - INTRODUCTION: Guidelines for warfarin management in children are essentially extrapolated from adult evidence. This study aimed to address that lack of paediatric-specific data regarding warfarin safety and efficacy for this population. MATERIALS AND METHODS: A retrospective clinical audit was conducted within a cardiac referral centre incorporating a paediatric anticoagulation service. Children (0-16 years) with a prosthetic cardiac valve were included. Warfarin related outcomes were collected between January 1st 2004 and December 31st 2009. Analysis included the percentage of INR tests within, above, or below the target therapeutic range (TTR). Adverse event data was collected regarding major bleeding and thrombotic events. RESULTS: 75 patient years of warfarin therapy were recorded. 44.0% of INR tests were within the TTR. INR tests not within the TTR were twice as likely to be sub-therapeutic. Children with aortic prosthetic valves achieved their TTR less frequently than children with prosthetic mitral or tricuspid valves. There were no thrombotic events and 3 major bleeding events. CONCLUSIONS: Although less than 50% of INR results were within the TTR, oral anticoagulant management resulted in acceptable safety and efficacy outcomes for this cohort. Further studies are needed to confirm optimal paediatric-specific warfarin management strategies for children with prosthetic heart valves. PMID- 21620443 TI - Antibodies to Domain I of beta(2)Glycoprotein I are in close relation to patients risk categories in Antiphospholipid Syndrome (APS). AB - INTRODUCTION: Antiphospholipid Syndrome (APS) is characterized by the presence of circulating antiphospholipid antibodies in patients with thrombosis or pregnancy morbidity. Antibodies involved in these disorders are mainly those directed against beta(2)-Glycoprotein I (beta(2)GPI) with the major epitope apparently located on discontinuous antigen with several parts of Domain I (DmI) involved. The relation between anti-DmI antibodies and patients' risk categories is unknown. MATERIALS AND METHODS: The synthetic full-length and correctly-folded DmI (1-64) to set up a competitive inhibition enzyme-linked immunoadsorbent assays (ELISA) was used. Plasma of 22 patients with APS and triple positivity [Lupus Anticoagulant positive (LAC+), IgG anti-cardiolipin positive (aCL+), IgG anti-beta(2)GPI positive (a beta(2)GPI +)], 15 with double positivity (IgG aCL+, IgG abeta(2)GPI+), 9 with single positivity (IgG abeta(2)GPI+) and 20 controls were evaluated. RESULTS: Median of percentage inhibition was 25.5% [interquartile range (IQR)17.2-33.0] in triple positive patients. Significantly lower inhibition was observed in patients with double positivity, median inhibition 5.0% (IQR 0.0 27.0) and in patients with single positivity median inhibition was 2.0% (IQR 0.5 8.0) (p<0.0001). No inhibition was detected in control subjects or using beta(2)GPI peptides (40-52 and 57-70), or when antithrombin, an insignificant control protein was used. CONCLUSIONS: High risk patients with APS and triple laboratory positivity as compared with double and single positivity patients have significantly higher titre of anti-DmI antibodies as evaluated by an inhibition test. PMID- 21620444 TI - The fate of donor osteocytes in fine particulate bone powders during repair of bone defects in experimental rats. AB - The aim of the study was to investigate the fate of donor osteocytes in fine particulate bone powders during repair of bone defects in experimental rats. The iliac bone of male inbred DA rats was harvested and used as the larger bone grafts and also prepared as fine particulate (granulated) bone powders (300 500MUm size particles) for transplantation into radial defects in female rats. The presence and relative amounts of genes specific to the sex-determining region of the Y-chromosome (Sry) originating from the bone grafts were evaluated by polymerase chain reaction and by in situ hybridization, respectively. Additional samples were evaluated histologically. In the larger bone grafts, the expression of Sry decreased relatively early, disappeared by 1 week, reappeared at 4 weeks and continued to increase with time. In the fine particulate bone powders, Sry was detected all the time and its expression was statistically greater than in the larger bone grafts at each time point. Both bone grafts provided donor cells to repair the defects. The donor cells seemed to function differently between the two groups. The fine particulate bone powders contained more living osteocytes in comparison with the larger bone grafts and may accelerate the healing of bone defects compared with conventional autografts. PMID- 21620445 TI - Steroid levels and the spatiotemporal expression of steroidogenic enzymes and androgen receptor in developing ovaries of immature rats. AB - Immunoexpression of 3beta-hydroxysteroid dehydrogenase (3beta-HSD), cytochrome P450c17 (P450c17), androgen receptor (AR), and steroid contents were studied in the ovaries of immature female Wistar rats killed between postnatal days 1 and 30. During days 1-7, ovarian somatic structures lacked AR, 3beta-HSD and P450c17, except for the surface epithelium, which featured the presence of these three proteins, suggestive of its androgen responsiveness and steroidogenic function. On day 10, AR appeared in many somatic structures, including the granulosa layers, which coincided with the P450c17 immunoexpression in some theca/interstitial cells, and an increase in ovarian androgen concentration. On the following days a further rise in ovarian androgen and progesterone contents paralleled an increase in 3beta-HSD and P450c17 immunoexpression in the theca layer cells and primary interstitial cells. However, the development of the follicles constituting the first follicular wave was aberrant, since they lacked AR expression until the preantral stage and were characterized by a delayed onset and much lower expression of the thecal P450c17. They could not ovulate, since ovarian content of estradiol was too low to evoke the LH surge. The clusters of the secondary interstitial cells found on day 30 exhibited predominant expression of 3beta-HSD over P450c17, suggesting more intensive progesterone than androgen synthesis in these structures. PMID- 21620446 TI - Impact of payer status on treatment of cervical cancer at a tertiary referral center. AB - OBJECTIVES: The study aims to determine the impact of payer status on the likelihood of receiving definitive treatment for invasive cervical cancer at a tertiary medical center. METHODS: All consecutive patients presenting to Johns Hopkins Hospital with a diagnosis of invasive cervical cancer between 1/1/95 12/31/08 were retrospectively identified from the tumor registry. Demographic and clinical information were abstracted from the medical record. Payer status was categorized as private, public, no insurance, or unknown. Treatment was defined as surgery, chemo-radiation, chemotherapy, radiation, or no definitive therapy. The likelihood of receiving no definitive therapy was analyzed using Pearson chi square analysis, univariate and multivariate models. RESULTS: A total of 306 patients were identified. Median age was 47 and 60% of patients had early stage disease at diagnosis (stages IA-IIA). Fifty-six percent of the cohort had private insurance, 34% had public insurance, and 6% had no insurance. Having no insurance was the single most significant risk factor associated with receiving no standard therapy. While 7% of privately insured and 4% of publicly insured patients did not receive definitive therapy, 16% of uninsured patients did not receive definitive treatment. In multivariate analysis controlling for age, race, stage, histology, and comorbidities, uninsured payer status was a significant and independent predictor of receiving no definitive treatment (OR 8.01, CI 1.265 50.694, p=0.027) than patients with public insurance. CONCLUSIONS: In this study, uninsured payer status was significantly associated with a higher likelihood of not receiving standard therapy for cervical cancer. Additional studies are warranted to characterize specific barriers to care for this at-risk population. PMID- 21620447 TI - Docetaxel plus trabectedin appears active in recurrent or persistent ovarian and primary peritoneal cancer after up to three prior regimes: a phase II study of the Gynecologic Oncology Group. PMID- 21620448 TI - Modulation of redox signaling promotes apoptosis in epithelial ovarian cancer cells. AB - OBJECTIVE: Epithelial ovarian cancer (EOC) cells are known to be resistant to apoptosis through a mechanism that may involve alteration in their redox balance. NADPH oxidase is a major source of intracellular superoxide, which is converted to the less toxic product by superoxide dismutase (SOD). Superoxide contributes to hypoxia inducible factor (HIF)-1alpha stabilization. We sought to determine the effects of inhibiting the generation of intracellular reactive oxygen species (ROS) on apoptosis of EOC cells. METHODS: Diphenyleneiodonium (DPI), an irreversible ROS inhibitor, was used to inhibit the generation of ROS in EOC cell lines, SKOV-3 and MDAH-2774, followed by assessment of apoptosis, NADPH oxidase, SOD3 and HIF-1alpha expression. A combination of immunohistochemistry, immunoprecipitation/western blot, and real-time RT-PCR were utilized to evaluate the expression of these enzymes in EOC cells as well as normal ovarian tissue and ovarian cancer tissue specimens. RESULTS: DPI treatment significantly induced apoptosis in both EOC cell lines as evident by increased caspase-3 activity and TUNEL assay. Additionally, both EOC cell lines were found to express NADPH oxidase, HIF-1alpha, and SOD3, which were highly sensitive to DPI treatment. DPI treatment resulted in reduced NADPH oxidase, SOD3 and HIF-1alpha levels. Furthermore, ovarian cancer tissues were found to manifest higher NADPH oxidase levels as compared to normal ovarian tissues. CONCLUSIONS: These data suggest that lowering oxidative stress, possibly through the inhibition of NADPH oxidase, induces apoptosis in ovarian cancer cells and may serve as a potential target for cancer therapy. PMID- 21620449 TI - Re: "Rate of para-aortic lymph node micrometastasis in patients with locally advanced cervical cancer". PMID- 21620450 TI - High-risk human papillomavirus seems not involved in DES-related and of limited importance in nonDES related clear-cell carcinoma of the cervix. AB - OBJECTIVE: Over 90% of all cervical adenocarcinoma are caused by a transforming infection with a high-risk type human papillomavirus (hrHPV). Previous studies demonstrated that the association between hrHPV positivity and cervical clear cell adenocarcinoma (CCAC) varies between 0% and 100%. As approximately 60% of all CCAC are associated with intra-uterine diethylstilbestrol (DES) exposure, we determined in a cohort of both DES-exposed and DES-unexposed women the prevalence of hrHPV infections, and the potential etiological role of hrHPV by additional analysis of p16INK4a and p53 expression. METHODS: Representative slides of 28 women diagnosed with CCAC were tested for hrHPV by two PCR methods (the clinically validated GP5+/6+ PCR and the very sensitive SPF(10)PCR/LiPA(25)). Fifteen women were DES-exposed, 10 unexposed and of 3 women DES-exposure was unknown. Twenty-one cases with sufficient material were immuno-histochemically stained for p16INK4a and p53. RESULTS: Seven tumors, of which four DES-exposed and two unexposed tested positive for hrHPV with GP5+/6+ PCR. Thirteen tumors, of which five DES-exposed and seven unexposed, tested positive with SPF(10)PCR/LiPA(25). In one women with unknown exposure, a CCAC tested positive in both assays. Only three cases, none in DES-exposed women, and all positive with both hrHPV assays, revealed diffuse p16INK4a immuno-staining and weak p53 staining as well, supporting indisputable hrHPV involvement. CONCLUSIONS: Although the prevalence of hrHPV was high, only two DES-unrelated CCAC (25%) and one tumor in a woman with unknown exposure could be attributed to hrHPV. PMID- 21620451 TI - Autoreactive CD4(+) T cells in patients with atopic dermatitis. PMID- 21620452 TI - Subcutaneous immunotherapy and pharmacotherapy in seasonal allergic rhinitis: a comparison based on meta-analyses. AB - BACKGROUND: Allergen-specific subcutaneous immunotherapy (SCIT) of seasonal allergic rhinitis (SAR) is usually considered a "second-line," slow-acting, disease-modifying treatment. OBJECTIVE: We sought to test whether SCIT is as effective as antisymptomatic treatment in the control of symptoms in patients with SAR in the first year of treatment. METHODS: We reviewed meta-analyses with 5 or more randomized, double-blind, placebo-controlled trials of SCIT or antisymptomatic treatment in patients with SAR. We then selected trials measuring the total nasal symptom score (TNSS), the total symptom score (TSS), or both during the first pollen season after treatment initiation. Efficacy was determined as the percentage reduction in TSSs and TNSSs obtained with active treatment compared with placebo (relative clinical impact [RCI]) and the standardized mean difference (SMD) of treatment verses placebo (effect size [ES]). RESULTS: The weighted mean RCI of SCIT on TNSSs (-34.7% +/- 6.8%) was higher than those of mometasone (-31.7% +/- 16.7%, P < .00001) and montelukast ( 6.3% +/- 3.0%, P < .00001). The weighted mean RCI of SCIT on TSSs (-32.9% +/- 12.7%) was higher than that of desloratadine (-12.0% +/- 5.1%, P < .00001). The overall ES of SCIT in terms of TNSSs (SMD, -0.94; 95% CI, -1.45 to -0.43) was similar to that of mometasone (SMD, -0.47; 95% CI, -0.63 to -0.32; P > .05) and higher than that of montelukast (SMD, -0.24; 95% CI, -0.33 to -0.16; P < .05). The overall ES of SCIT in terms of TSSs (SMD, -0.86; 95% CI, -1.17 to -0.55) was comparable with that of desloratadine (SMD, -1.00; 95% CI, -1.68 to -0.32; P > .05). CONCLUSIONS: Our data provide indirect but consistent evidence that SCIT is at least as potent as pharmacotherapy in controlling the symptoms of SAR as early as the first season of treatment. PMID- 21620453 TI - Centriolar kinesin Kif24 interacts with CP110 to remodel microtubules and regulate ciliogenesis. AB - We have identified a protein, Kif24, that shares homology with the kinesin-13 subfamily of motor proteins and specifically interacts with CP110 and Cep97, centrosomal proteins that play a role in regulating centriolar length and ciliogenesis. Kif24 preferentially localizes to mother centrioles. Loss of Kif24 from cycling cells resulted in aberrant cilia assembly but did not promote growth of abnormally long centrioles, unlike CP110 and Cep97 depletion. We found that loss of Kif24 leads to the disappearance of CP110 from mother centrioles, specifically in cycling cells able to form cilia. Kif24 is able to bind and depolymerize microtubules in vitro. Remarkably, ectopically expressed Kif24 specifically remodels centriolar microtubules without significantly altering cytoplasmic microtubules. Thus, our studies have identified a centriolar kinesin that specifically remodels a subset of microtubules, thereby regulating cilia assembly. These studies also suggest mechanistic differences between the regulation of microtubule elongation associated with centrioles and cilia. PMID- 21620454 TI - Aquaporin inhibition changes protein phosphorylation pattern following sperm motility activation in fish. AB - Our previous studies demonstrated that osmolality is the key signal in sperm motility activation in Sparus aurata spermatozoa. In particular, we have proposed that the hyper-osmotic shock triggers water efflux from spermatozoa via aquaporins. This water efflux determines the cell volume reduction and, in turn, the rise in the intracellular concentration of ions. This increase could lead to the activation of adenylyl cyclase and of the cAMP-signaling pathway, causing the phosphorylation of sperm proteins and then the initiation of sperm motility. This study confirms the important role of sea bream AQPs (Aqp1a and Aqp10b) in the beginning of sperm motility. In fact, when these proteins are inhibited by HgCl(2), the phosphorylation of some proteins (174 kDa protein of head; 147, 97 and 33 kDa proteins of flagella), following the hyper-osmotic shock, was inhibited (totally or partially). However, our results also suggest that more than one transduction pathways could be activated when sea bream spermatozoa were ejaculated in seawater, since numerous proteins showed an HgCl(2)(AQPs) independent phosphorylation state after motility activation. The role played by each different signal transduction pathways need to be clarified. PMID- 21620455 TI - The neural basis of obligatory decomposition of suffixed words. AB - Recent neurolinguistic studies present somewhat conflicting evidence concerning the role of the inferior temporal cortex (IT) in visual word recognition within the first 200 ms after presentation. On the one hand, fMRI studies of the Visual Word Form Area (VWFA) suggest that the IT might recover representations of the orthographic form of words. On the other hand, influential MEG studies of responses from the occipito-temporal regions around 150 ms post-stimulus onset indicate recognition of letters as opposed to symbols but not a sensitivity to statistical properties of letter strings associated with word form representations. Recent MEG experiments support the position that the IT does represent the visual word forms of morphemes and performs morphological decomposition modulated by the statistical relations between morphemes by 170 ms post presentation (at the M170 response). Responses to heteronyms show that the M170 does not make contact with the mental lexicon where word forms are connected to meanings. We report here an MEG study of pseudo-affixed words like brother, which masked priming studies have shown are decomposed in recognition. If the M170 response from IT does index obligatory morphological decomposition based on visual word forms but not lexical entries, we should find that the statistical relation between pseudo-stem and pseudo-suffix modulates the M170 for pseudo affixed words, as for truly affixed words. The results of this experiment confirm this prediction. In addition, surface form frequency for these words also modulates the M170, providing some support for dual route recognition for words for which decomposition is a garden path. PMID- 21620456 TI - Visual field differences in visual word recognition can emerge purely from perceptual learning: evidence from modeling Chinese character pronunciation. AB - In Chinese orthography, a dominant character structure exists in which a semantic radical appears on the left and a phonetic radical on the right (SP characters); a minority opposite arrangement also exists (PS characters). As the number of phonetic radical types is much greater than semantic radical types, in SP characters the information is skewed to the right, whereas in PS characters it is skewed to the left. Through training a computational model for SP and PS character recognition that takes into account of the locations in which the characters appear in the visual field during learning, but does not assume any fundamental hemispheric processing difference, we show that visual field differences can emerge as a consequence of the fundamental structural differences in information between SP and PS characters, as opposed to the fundamental processing differences between the two hemispheres. This modeling result is also consistent with behavioral naming performance. This work provides strong evidence that perceptual learning, i.e., the information structure of word stimuli to which the readers have long been exposed, is one of the factors that accounts for hemispheric asymmetry effects in visual word recognition. PMID- 21620457 TI - Ethical issues of hospital crowding solutions. PMID- 21620458 TI - Child occupants and side-impact crashes: commentary. PMID- 21620461 TI - Intravenous HYDROmorphone: can you manage the risk? PMID- 21620463 TI - Evolution of a triage and flow process during a joint American-Chinese medical mission in Dali, Yunnan, China. PMID- 21620464 TI - Perceived parental psychological control and adolescent depressive experiences: a cross-cultural study with Belgian and South-Korean adolescents. AB - In recent research on psychologically controlling parenting, debate has arisen about the cross-cultural relevance of this construct, with some scholars arguing that the developmental outcomes of psychological control are culture-bound and others arguing that the detrimental effects of psychological control generalize across cultures. This study aimed to add to this debate by examining the relevance of a distinction between two domain-specific expressions of psychological control (i.e., dependency-oriented and achievement-oriented) in Belgian (N = 290) and South-Korean (N = 321) adolescents. Multi-group path analyses showed that associations between the domains of psychological control, depressive personality, and depressive symptoms were similar between the two samples. Overall, the findings are in line with the notion that the effects of psychological control generalize across culture. PMID- 21620465 TI - Brief report: gum chewing affects standardized math scores in adolescents. AB - Gum chewing has been shown to improve cognitive performance in adults; however, gum chewing has not been evaluated in children. This study examined the effects of gum chewing on standardized test scores and class grades of eighth grade math students. Math classes were randomized to a gum chewing (GC) condition that provided students with gum during class and testing, or a control condition with no gum (NGC). Participants included 108 students. The math sections of the Texas Assessment of Knowledge and Skills (TAKS) and the Woodcock Johnson III Tests of Achievement (WJ-III), and math class grades were used to assess academic performance. Students in the gum chewing condition improved standardized test scores and maintained higher grades in math class compared to those in the no-gum chewing condition. These results are encouraging as gum chewing may be a cost effective and easily implemented method to increase student performance. TRIAL REGISTRATION: NCT00792116. PMID- 21620466 TI - Pancreatic cancer. AB - Substantial progress has been made in our understanding of the biology of pancreatic cancer, and advances in patients' management have also taken place. Evidence is beginning to show that screening first-degree relatives of individuals with several family members affected by pancreatic cancer can identify non-invasive precursors of this malignant disease. The incidence of and number of deaths caused by pancreatic tumours have been gradually rising, even as incidence and mortality of other common cancers have been declining. Despite developments in detection and management of pancreatic cancer, only about 4% of patients will live 5 years after diagnosis. Survival is better for those with malignant disease localised to the pancreas, because surgical resection at present offers the only chance of cure. Unfortunately, 80-85% of patients present with advanced unresectable disease. Furthermore, pancreatic cancer responds poorly to most chemotherapeutic agents. Hence, we need to understand the biological mechanisms that contribute to development and progression of pancreatic tumours. In this Seminar we will discuss the most common and deadly form of pancreatic cancer, pancreatic ductal adenocarcinoma. PMID- 21620467 TI - 5 versus 10 days of treatment with ceftriaxone for bacterial meningitis in children: a double-blind randomised equivalence study. AB - BACKGROUND: Bacterial meningitis is an important cause of morbidity and mortality in developing countries, but the duration of treatment is not well established. We aimed to compare the efficacy of 5 and 10 days of parenteral ceftriaxone for the treatment of bacterial meningitis in children. METHODS: We did a multicountry, double-blind, placebo-controlled, randomised equivalence study of 5 versus 10 days of treatment with ceftriaxone in children aged 2 months to 12 years with purulent meningitis caused by Streptococcus pneumoniae, Haemophilus influenzae type B, or Neisseria meningitidis. Our study was done in ten paediatric referral hospitals in Bangladesh, Egypt, Malawi, Pakistan, and Vietnam. We randomly assigned children who were stable after 5 days of treatment, through site-balanced computer-generated allocation lists, to receive a further 5 days of ceftriaxone or placebo. Patients, their guardians, and staff were masked to study-group allocation. Our primary outcomes were bacteriological failure or relapse. Our analysis was per protocol. This study is registered with the International Standard Randomised Controlled Trial Number Register, number ISRCTN38717320. FINDINGS: We included 1004 of 1027 children randomly assigned to study groups in our analyses; 496 received treatment with ceftriaxone for 5 days, and 508 for 10 days. In the 5-day treatment group, two children (one infected with HIV) had a relapse; there were no relapses in the 10-day treatment group and there were no bacteriological failures in either study group. Side-effects of antibiotic treatment were minor and similar in both groups. INTERPRETATION: In children beyond the neonatal age-group with purulent meningitis caused by S pneumoniae, H influenzae type b, or N meningitidis who are stable by day 5 of ceftriaxone treatment, the antibiotic can be safely discontinued. FUNDING: United States Agency for International Development. PMID- 21620468 TI - Fine with five? Shorter antibiotic courses for childhood meningitis. PMID- 21620469 TI - Establishment of self-organization system in rapidly formed multicellular heterospheroids. AB - Multicellular heterospheroids including two or more cell types have some tissue/organ properties and can be used in cell-to-cell interaction studies. However, the spheroid formation is difficult to control because the adhesion efficacy is different in each cell type. To solve this, we applied a rapid cell to-cell adhesion method, avidin-biotin (AB) binding, to spheroid formation. Introduction of avidin or biotin molecules to the cell surfaces of Mile Sven 1 (MS1) cells promoted formation of spheroid in minutes. This method allowed the construction of heterospheroids having homogenous distributions of different cell types. Interestingly, cells showed self-organization and MS1 cells formed networks with Hep G2 cells. NIH3T3 cells also remodeled when mixed with Hep G2 cells. In contrast, a combination of MS1 and NIH3T3 cells failed to show pattern formation, indicating that self-organization was based on the composition of cell types. Actin polymerization not cell proliferation was the dominant factor in remodeling of heterospheroids in the first 24 h. We also demonstrated the self organization of spheroids comprising three different cell types. The new technology to assemble cells is important not only to study cell-to-cell interaction but also to make three-dimensional complicated tissues. PMID- 21620470 TI - Simultaneous in vivo tracking of dendritic cells and priming of an antigen specific immune response. AB - We report the fabrication of a one-pot antigen system that delivers antigen to dendritic cells (DCs) and tracks their in vivo migration after injection. Multifunctional polymer nanoparticles containing ovalbumin protein, magnetic resonance imaging contrast agents (iron oxide nanoparticles), and near-infrared fluorophores (indocyanine green, ICG), MPN-OVA, were prepared using a double emulsion method. The MPN-OVA was efficiently taken up by the dendritic cells and subsequently localized in the lysosome. Flow cytometry analysis revealed an increase in the uptake of OVA antigen by MPN-OVA at 37 degrees C, when compared with soluble OVA protein. We found that MPN-OVA had no effect on DC surface expression of MHC class I, costimulatory (CD80, CD86) or adhesion (CD54) molecules or the ability of DCs to mature in response to LPS. Following the uptake of MPN-OVA, exogenous OVA antigen was delivered to the cytoplasm, and OVA peptides were presented on MHC class I molecules, which enhanced OVA antigen specific cross-presentation to OT-1 T cells and CD8OVA1.3 T cell hybridoma in vitro. The immunization of mice with MPN-OVA-treated DCs induced OVA-specific CTL activity in draining lymph nodes. The presence of MPN allowed us to monitor the migration of DCs via lymphatic drainage using NIR fluorescence imaging, and the homing of DCs into the lymph nodes was imaged using MRI. This system has potential for use as a delivery system to induce T cell priming and to image DC based immunotherapies. PMID- 21620471 TI - Large-scale production of murine embryonic stem cell-derived osteoblasts and chondrocytes on microcarriers in serum-free media. AB - The generation of tissue-engineered constructs from stem cells for the treatment of musculoskeletal diseases may have immense impact in regenerative medicine, but there are difficulties associated with stem cell culture and differentiation, including the use of serum. Here we present serum-free protocols for the successful production of murine embryonic stem cell (mESC) derived osteoblasts and chondrocytes on CultiSpher S macroporous microcarriers in stirred suspension bioreactors. Various inoculum forms and agitation rates were investigated. Produced osteogenic cells were implanted ectopically into SCID mice and orthotopically into a murine burr-hole fracture model. Osterix, osteocalcin and collagen type I were upregulated in osteogenic cultures, while aggrecan and collagen type II were upregulated in chondrogenic cultures. Histological analysis using alizarin red S, von Kossa and alcian blue staining confirmed the presence of osteoblasts and chondrocytes, respectively in cultured microcarriers and excised tissue. Finally, implantation of derived cells into a mouse fracture model revealed cellular integration without any tumor formation. Overall, microcarriers may provide a supportive scaffold for ESC expansion and differentiation in a serum-free bioprocess for in vivo implantation. These findings lay the groundwork for the development of clinical therapies for musculoskeletal injuries and diseases using hESCs and iPS cells. PMID- 21620472 TI - The t(11;19)(q23;p13) fusing MLL with MYO1F is recurrent in infant acute myeloid leukemias. PMID- 21620473 TI - Levels of household particulate matter and environmental tobacco smoke exposure in the first year of life for a cohort at risk for asthma in urban Syracuse, NY. AB - The Syracuse, NY, AUDIT (Assessment of Urban Dwellings for Indoor Toxics) study was designed to quantify asthma agent levels in the inner-city homes of a birth cohort whose mothers had a diagnosis of asthma. Risk of exposure to particulate matter (PM), particle number and tobacco smoke was assessed in 103 infants' homes. Repeat measurements were made in 44% of the homes. Infants also were examined on a quarterly basis during the first year of life to monitor their respiratory health and urine cotinine levels. Overall geometric mean (GM) values for PM(2.5) of 21.2 MUg/m(3) and for PM(10) of 31.8 MUg/m(3) were recorded in homes at visit 1. GM values for PM(2.5) and PM(10) in smoking homes were higher at 26.3 and 37.7 MUg/m(3), while values in non-smoking homes were 12.7 and 21.2 MUg/m(3) respectively. Fifty-four percent of mothers (55/103) smoked at some point in pregnancy (39% smoked throughout pregnancy). Environmental tobacco smoke (ETS) exposure occurred in 68% of homes during the infants' first year. Significant to this study was the size- and time-resolved monitoring of PM at 140 home visits and the classification of PM count data. PM number counts ranged from continuously low levels (little indoor activity) to continuously high counts (constant indoor activity), and recorded apparent instances of prolonged repeated cigarette smoking. Wheezing in the first year of life was recorded for 38% of the infants (39/103). Adjusted logistic regression modeling demonstrated that elevated levels of indoor PM(2.5) (>= 15 MUg/m(3)) were a significant risk factor for infant wheezing after controlling for infant gender, mothers' age and education level, season of home visit and presence of carpeting (OR 4.21; 95% CI 1.36-13.03; p=0.013). An elevated level of the nicotine metabolite cotinine in infant urine also was associated with infant wheezing after adjusting for infant gender, mothers' age and education level (OR 5.10; 95% CI 0.96-27.24; p=0.057). ETS exposure was pervasive in the AUDIT cohort and a risk for developing infants in this urban population. PMID- 21620474 TI - Comparison on gestation and lactation exposure of perfluorinated compounds for newborns. AB - Perfluorinated compounds (PFCs) are worldwide present in the environment and the general population. Animal studies have shown developmental toxicity of these compounds. To investigate the PFCs exposure of neonates from mother during gestation and lactation, we analyzed twelve PFCs in matched maternal serum, cord serum and breast milk samples collected from 50 pairs of women and their newborns between June and July 2009 in Jinhu, China. Eight PFCs were detected in serum samples, and five of them were also detectable in breast milk. A significant intercorrelation between PFCs concentrations in matched maternal serum, cord serum and breast milk was observed (p<0.01, r=0.435-0.911). The median partition ratio was from 0.39:1 (PFDA) to 1.74:1 (PFTrDA) for seven PFCs through the placenta, and was from 0.02:1 (PFOS) to 0.09:1 (PFOA) for five PFCs through the lactation. A high transport efficiency of PFOA both through placental barrier and lactation was observed. The postnatal exposure of PFCs through lactation was higher compared to prenatal exposure, especially for PFOA. PMID- 21620475 TI - Smoking, dietary betaine, methionine, and vitamin D in monozygotic twins with discordant macular degeneration: epigenetic implications. AB - OBJECTIVE: We evaluated monozygotic twin pairs with discordant age-related macular degeneration (AMD) phenotypes to assess differences in behavioral and nutritional factors. DESIGN: Case series. PARTICIPANTS: Caucasian male twin pairs from the United States Twin Study of Macular Degeneration. METHODS: Twin pairs were genotyped to confirm monozygosity. Ocular characteristics were evaluated based on fundus photographs using the Wisconsin Grading System and a 5-grade Clinical Age-Related Maculopathy Staging System. We selected twin pairs discordant in each of the following phenotypic categories: Stage of AMD (n = 28), drusen area (n = 60), drusen size (n = 40), and increased pigment area (n = 56). The Wilcoxon signed-rank test and linear regression were used to assess associations between behavioral and nutritional characteristics and each phenotype within discordant twin pairs. MAIN OUTCOME MEASURES: Differences in smoking and dietary factors within twin pairs discordant for stage of AMD, drusen area, drusen size, and pigment area. RESULTS: Representative fundus photographs depict the discordant phenotypes. Pack-years of smoking were higher for the twin with the more advanced stage of AMD (P = 0.05). Higher dietary intake of vitamin D was present in the twins with less severe AMD (P = 0.01) and smaller drusen size (P = 0.05) compared with co-twins, adjusted for smoking and age. Dietary intakes of betaine and methionine were significantly higher in the twin with lower stage of AMD (P = 0.009) and smaller drusen area (P = 0.03), respectively. CONCLUSIONS: The twin with the more advanced stage of AMD, larger drusen area, drusen size, and pigment area tended to be the heavier smoker. The twin with the earlier stage of AMD, smaller drusen size and area, and less pigment tended to have higher dietary vitamin D, betaine, or methionine intake. Results suggest that behavioral and nutritional factors associated with epigenetic mechanisms are involved in the etiology of AMD, in addition to genetic susceptibility. PMID- 21620476 TI - Attitudes about depression and its treatment among mental health professionals, lay persons and immigrants and refugees in Norway. AB - BACKGROUND: Internationally, depression is a common psychological disorder whose treatment depends upon its identification by treating professionals as well as patient utilization of mental health care systems; the latter often being hampered by cultural differences between patients and health professionals. METHOD: The current study used vignettes of depressed patients which varied the culture and/or social circumstances of the patient to assess whether these variables influenced the conceptualization of depression and its treatment. Participants (N=722) included mental health professionals, lay people, immigrants, and refugees in Norway. RESULTS: We found that immigrants and refugees, particularly those of non-western origin, endorsed different types of depression treatments from native Norwegians and mental health professionals, and judged who deserved treatment and who was overreacting based on the patient's culture and social circumstances, while native Norwegians did not. LIMITATIONS: While widely used cross-culturally, vignette methodology is limited in its generalizability to real clinical situations. Acculturation was not evaluated, which may have influenced the results. CONCLUSIONS: Findings support the integration of cultural competency ideals not only into treatment, but also into public health promotions of mental health services for lay people. PMID- 21620478 TI - Complementary medicine, self-help, and lifestyle interventions for obsessive compulsive disorder (OCD) and the OCD spectrum: a systematic review. AB - BACKGROUND: In Obsessive Compulsive Disorder (OCD) current standard pharmacotherapies may be of limited efficacy. Non-conventional interventions such as Complementary and Alternative Medicine (CAM), self-help techniques, and lifestyle interventions are commonly used by sufferers of OCD, however to date no systematic review of this specific area exists. METHODS: We conducted a systematic review of studies using CAM, self-help, and lifestyle interventions for treatment of OCD and trichotillomania (TTM). PubMed, PsycINFO, China Academic Journals Full-text Database, The Cochrane Library and CINAHL were searched (up to Jan 11th 2011), for controlled clinical trials using non-conventional interventions for OCD. A quality analysis using a purpose-designed scale and an estimation of effect sizes (Cohen's d) where data was available, were also calculated. RESULTS: The literature search revealed 14 studies that met inclusion criteria. Methodological quality of nutraceutical studies (nutrients and herbal medicines) were rated as high (mean 8.6/10), whereas mind-body or self-help studies were poorer (mean 6.1/10). In OCD, tentative evidentiary support from methodologically weak studies was found for mindfulness meditation (d=0.63), electroacupuncture (d=1.16), and kundalini yoga (d=1.61). Better designed studies using the nutrient glycine (d=1.10), and traditional herbal medicines milk thistle (insufficient data for calculating d) and borage (d=1.67) also revealed positive results. A rigorous study showed that N-acetylcysteine (d=1.31) was effective in TTM, while self-help technique "movement decoupling" also demonstrated efficacy (d=0.94). Mixed evidence was found for myo-inositol (mean d=0.98). Controlled studies suggest that St John's wort, EPA, and meridian tapping are ineffective in treating OCD. CONCLUSIONS: While several studies were positive, these were un-replicated and commonly used small samples. This precludes firm confidence in the strength of clinical effect. Preliminary evidence however is encouraging, and more rigorous research of some of the more hypothesis-based interventions in the treatment of OCD and TTM may be indicated. PMID- 21620477 TI - Neurocognitive impairment in adolescent major depressive disorder: state vs. trait illness markers. AB - BACKGROUND: Current treatment outcomes of Major Depressive Disorder (MDD) in adolescents remain suboptimal. Discriminating between state and trait markers of MDD in adolescents would help identify markers that may guide choice of appropriate interventions and help improve longer-term outcome for individuals with the illness. METHODS: We compared neurocognitive performance in executive function, sustained attention and short-term memory in 20 adolescents with MDD in acute episode (MDDa), 20 previously depressed adolescents in remission (MDDr) and 17 healthy control participants (HC). RESULTS: There was a group difference that emerged for executive function with increasing task difficulty (p=0.033). MDDa showed impaired executive function, as measured by using more moves to solve 4 move problems on a forward planning task, relative to MDDr and HC (p=0.01, d=0.94 and p=0.015, d=0.77 respectively). MDDa showed more impulsivity as measured by lower response bias (B") on a sustained attention task than both MDDr and HC (p=0.01, d=0.85 and p=0.008, d=0.49 respectively). Higher impulsivity was associated with more severe depression (r=-0.365, p=0.022) and earlier age of onset of depression (r=0.402, p=0.012) and there was a trend for a correlation between more executive dysfunction and more severe depression (r=0.301 p=0.059) in MDDa and MDDr combined. The three groups did not differ significantly on short term memory or target detection on the sustained attention task. LIMITATION: These results need to be replicated in the future with a larger sample size. CONCLUSION: Executive dysfunction and impulsivity appear to be state-specific markers of MDD in adolescents that are related to depression severity and not present in remission. PMID- 21620479 TI - Tryptophan hydroxylase 2 gene is associated with major depressive disorder in a female Chinese population. AB - BACKGROUND: Previous candidate gene studies of major depressive disorder (MDD) have provided inconclusive evidence of association for genes with strong biological rationale for MDD. In this study, we aimed to investigate the association of tryptophan hydroxylase 2 gene with MDD and its treatment response in the Chinese Han population. METHODS: Three hundred and sixty eight depressed patients who met DSM-IV criteria for major depressive disorder were recruited for the study. 371 normal controls were recruited from local community. Patients and normal controls were genotyped for TPH2 (rs4290270 and rs7305115) variants by polymerase chain reaction. Male and female subjects were analyzed separately. RESULTS: No differences were found in the frequencies of the single alleles and genotypes of the tested polymorphisms between MDD patients and normal group. The frequency of the A-A haplotype was significantly higher in female MDD compared to healthy female controls (P<0.05). No significant association with treatment response was discovered in haplotype and single-marker analysis. LIMITATIONS: This study lacks a placebo control and we cannot definitively exclude the possibility that some patients in the responder group responded to the placebo effect alone. CONCLUSION: The result suggests that TPH2 gene may have a gender dependent effect on susceptibility to MDD but not with its treatment response in Chinese Han population. Further studies are needed to replicate the association that we observed. PMID- 21620480 TI - Gender differences in a cohort study of 604 bipolar patients: the role of predominant polarity. AB - BACKGROUND: Some clinical differences between gender regarding the course and outcome of bipolar disorders have already been described and some others remain still controversial. AIMS: To explore gender differences regarding clinical and socio-demographic characteristics amongst bipolar patients with particular attention to predominant polarity and depressive symptoms. METHOD: Data were collected from DSM-IV type I and II bipolar patients (n=604), resulting from the systematic follow-up of the Bipolar Disorders Program, Hospital Clinic of Barcelona, over an average follow-up of 10 years. Socio-demographic and clinical variables were collected in order to detect gender-related differences. RESULTS: Bipolar women are more likely than men to show a predominance of depressive polarity as well as a depressive onset whilst men would be more likely to suffer from comorbid substance use disorders. Women significantly have a higher lifetime prevalence of psychotic depression and a higher prevalence of axis II comorbid disorders. Bipolar women are also more likely to have a family history of suicide and a lifetime history of attempted suicide. Suicide attempts are more often violent amongst bipolar men. In a backward logistic regression model, two variables were responsible for most gender-related clinical differences: type of predominant polarity - more likely to be depressive amongst women - (B=-0.794, p=0.027, Exp(B)=0.452; CI= 0.223-0.915), alcohol abuse (B=-1.095, p=0.000, Exp(B)=2990; CI= 1.817-4.919) and cocaine abuse (B=0.784, p=0.033, Exp(B)=2.189; CI= 1.066-4.496) - more prevalent amongst men. CONCLUSION: The main characteristic featuring bipolar women is depression, both at illness onset and as a predominant polarity all along the illness course. This may have important diagnostic and therapeutic implications. PMID- 21620481 TI - The Diagnostic Interview of Children and Adolescents for Parents of Preschool and Young Children: psychometric properties in the general population. AB - There is a need for reliable and well-validated diagnostic measures for studying psychopathology in preschool and young children. The goal is to study the psychometric properties of the Diagnostic Interview for Children and Adolescents for Parents of Preschool and Young Children (DICA-PPYC) in the general population. A sample of 852 Spanish school children, aged 3 to 7 years, were randomly selected and screened for a double phase design. A total of 251 families were interviewed with the DICA-PPYC and 244 participated in a test-retest design. Different measures of psychopathology and functional impairment were also administered. Test-retest agreement with a mean interval of 8.8 days ranged from excellent to slight (kappa from 1 to 0.39) for DSM-IV-TR and from good to fair (kappa from 0.77 to 0.49) for Research Diagnostic Criteria-Preschool Age diagnoses. Attenuation between test and retest was not significant for the prevalence of diagnoses, although it was significant for the number of externalising and total symptoms in the interview. The diagnoses converged moderately with the CBCL and Dominic scores. The presence of diagnoses in the DICA-PPYC significantly differentiated preschoolers and young children who had used mental health services, were more impaired, and presented more severe psychopathology measured by dimensional scales. The DICA-PPYC is a reliable and valid semi-structured interview schedule for preschool and young children, and can serve to advance the knowledge and mental health care of this population. PMID- 21620482 TI - The C(-1562)T polymorphism of matrix metalloproteinase-9 gene is associated with schizophrenia in China. PMID- 21620483 TI - Early predictors of posttraumatic stress in accident victims. AB - This study investigated the predictive value of pretraumatic and peritraumatic variables and early PTSD-like symptoms (within 48 h post-event) in determining the severity of posttraumatic stress in accident victims. Symptom development appears to be related to initial PTSD-like reactions, while demographic variables play a role in predicting symptom severity 6 months post-event. PMID- 21620484 TI - Theory of mind and facial emotion recognition in euthymic bipolar I and bipolar II disorders. AB - The main aim of this study was to compare patients with euthymic bipolar I (BDI) and bipolar II (BDII) disorders and healthy controls in measures of social cognition. Additional aims were to explore the association between social cognition performance with neurocognitive impairments and psychosocial functioning. Eighty one euthymic patients with BDI or BDII and 34 healthy controls were included. All subjects completed tests to assess verbal memory, attention, and executive functions. Additionally theory of mind (ToM) and facial emotion recognition measures were included. Psychosocial functioning was assessed with the GAF. Both groups of patients had lower performance than healthy controls in ToM, and a lower recognition of fear facial expression. When neurocognitive impairments and exposure to medications were controlled, performance in ToM and recognition of fear facial expression did not allow predicting if a subject was patient or healthy control. Social cognition measures not enhance variance beyond explained by neurocognitive impairments and they were not independent predictors of psychosocial functioning. Impairments in facial emotion recognition and ToM are mediated, at least partly, by attention-executive functions deficits and exposure to psychotropic medications. Likewise, social cognition measures did not contribute to variance beyond neurocognitive impairments. PMID- 21620485 TI - Amyloid beta 1-42 in cerebrospinal fluid is associated with cognitive plasticity. AB - Cerebrospinal fluid (CSF) total tau-protein (t-tau) and amyloid-beta 1-42 (Abeta(1-42)) have been increasingly included in the diagnostic process of Alzheimer's disease (AD). We aimed to analyze whether these CSF biomarkers correlate with cognitive plasticity as measured by a dynamic recognition test strategy. We assessed 29 elderly individuals (15 with incipient and 14 without AD) from an outpatient memory clinic at a university hospital by a Testing-the Limits (TtL) based recognition paradigm consisting of a pre-test (baseline) and two post-test conditions with an interposed encoding instruction. We identified a negative association between Abeta(1-42) and the two post-test failure rates, but not with that of the pre-test. Also, none of the standard tests correlates with Abeta 42-1 level. T-tau does not correlate with recognition performance. Our results suggest that Abeta(1-42) could be useful as a state marker for cognitive plasticity. PMID- 21620486 TI - Differential conditioned fear response predicts duloxetine treatment outcome in male veterans with PTSD: a pilot study. AB - This pilot study tested whether posttraumatic stress disorder (PTSD) patients with impaired conditioned fear acquisition were refractory to open-label duloxetine treatment. Patients with a differential conditioned fear response at pre-treatment subsequently demonstrated significant reductions in PTSD symptoms. These data provide initial evidence of a putative biomarker of selective treatment response in PTSD. PMID- 21620487 TI - Progress of the comparative MHC Committee and a summary of the Comparative MHC Workshops held at the 32nd ISAG, Edinburgh and the 9th IVIS, Tokyo, 2010. AB - The comparative MHC Nomenclature Committee was formed following a recommendation by the International Society for Animal Genetics (ISAG) at its meeting in Gottingen, Germany in 2002. The committee subsequently became affiliated to VIC, the Veterinary Immunology Committee of IUIS; the International Union of Immunological Societies, which is the umbrella organization for immunology societies worldwide. The committee currently fulfils three roles: (1) to establish guidelines for MHC nomenclature in species outside of human and mouse, (2) to facilitate the development of databases which provide a source of validated MHC sequences with official nomenclatures for associated species (immuno-polymorphism database IPD-MHC) and (3) to facilitate scientific discussion on comparative aspects of MHC biology. The committee currently comprises individuals representing a range of species nomenclature committees. This report summarises progress the committee has made in addressing its core objectives as well as the principal outputs from the Comparative MHC Workshops held at the 32nd International Society for Animal Genetics (ISAG) conference in Edinburgh, UK in July 2010 as well as in August 2010 at the 9th International Veterinary Immunology Symposium (IVIS) in Tokyo, Japan. Additional data from posters and presentations at related workshops focussing on aspects of MHC biology are also included where appropriate. PMID- 21620488 TI - Rhipicephalus (Boophilus) microplus embryo proteins as target for tick vaccine. AB - Rhipicephalus (Boophilus) microplus is one of the most widely distributed tick in the world. The control of the parasite is based mainly on the use of chemical acaricides, which are produced from a limited set of molecules. These drugs induce selection of acaricide-resistant ticks, and are an important source of environmental pollution. An approach based on anti-tick vaccines may circumvent these obstacles. Characterization of the physiological function of tick molecules may be useful to develop new vaccines. Previously, we reported the ability of some tick proteins as inducers of protective immune response. Vaccination studies using tick proteins like native (nBYC), recombinant (rBYC) egg-yolk aspartic endopeptidase and cysteine endopeptidase (VTDCE) from R. microplus and glutathione S-transferase (Hl-GST) from Haemaphysalis longicornis demonstrated the immunogenicity and antigenicity of these proteins in bovines. Eventually, immunization with these proteins triggered a partial immune response against R. microplus infestation in cattle, manifested mainly as a reduction in egg fertility (7.7% and 13.9% for nBYC, 5.9% for rBYC; 4.7% for VTDCE, 7.9% for Hl GST), and in the number of fully engorged ticks (18.2% for rBYC, 14.6% for VTDCE, 53% for Hl-GST). The data so far obtained suggest that these proteins have potential to be used as antigens in an anti-tick vaccine. Other proteins involved in tick embryogenesis also have this potential, like THAP and BmCl1, which are enzymes with key roles in vitellin and hemoglobin hydrolysis. Moreover, the identification of analogous proteins present in other tick species may bring information about the way to develop a vaccine against multiple tick species which can help to solve the problem faced by numerous countries where animals are parasitized by more than one tick species. The aim of the present review is to comprehensibly summarize the data obtained in the last few years by our collaborative research, discussing the efforts we have made to find antigens efficient enough for a cattle tick-controlling vaccine. This review discusses tick physiology studies aimed at the selection of possible targets, characterization of the selected proteins with emphasis on their biochemical and immunological aspects and results of vaccine trials on bovines. PMID- 21620489 TI - Combined chemotherapy with cytotoxic and targeted compounds for the management of human malignant pleural mesothelioma. AB - Human malignant pleural mesothelioma (hMPM) is an aggressive asbestos-associated cancer, the incidence of which is increasing and which, despite progress in diagnosis and therapy, continues to have a poor prognosis. Asbestos fibers induce aberrant cell signaling, leading to proto-oncogene activation and chemoresistance. In this review, we discuss the evolution of pharmacological management of hMPM up to the most recent advances. Monotherapy with single cytotoxic drugs achieves modest objective response rates, seldom reaching 30%. However, combination regimens using novel drugs and standard molecules are showing gradually improving responses and clinical benefits. Phase II/III studies have identified pemetrexed, a multitarget folate pathway inhibitor in combination with platinum derivatives, and the cisplatin/gemcitabine association as front line chemotherapy for hMPM. Detailed knowledge of molecular mechanisms of signal transduction and neoangiogenesis in hMPM should aid in the design and screening of other promising compounds such as more efficacious receptor tyrosine kinase inhibitors. PMID- 21620490 TI - Enhanced protection against pulmonary hypertension with sildenafil and endothelial progenitor cell in rats. AB - BACKGROUND: Sildenafil and bone marrow-derived endothelial progenitor cells (BMDEPCs) have been shown to ameliorate monocrotaline (MCT)-induced pulmonary arterial hypertension (PAH) in the rat. We test whether combined sildenafil and BMDEPC treatment exerts additional protection against MCT-induced PAH in rats. METHODS: Male Sprague-Dawley rats were randomized to receive saline injection only (group 1), MCT (70 mg/kg) only (group 2), MCT plus autologous BMDEPC (2.0*10(6) cells) transplantation (group 3), MCT with sildenafil (30 mg/kg/day) (group 4), and MCT with combined BMDEPCs-sildenafil (30 mg/kg/day) (group 5). Intravenous BMDEPC and oral sildenafil were given on day 3 after MCT administration. Hemodynamics were analyzed using Labchart software, whereas cellular and molecular parameters were measured using flow cytometry, real-time PCR, TUNEL assay, Western blot, and immunohistochemical staining. RESULTS: By day 35 following MCT treatment, lower expression of connexin43, protein kinase C epsilon, Bcl-2, and endothelial nitric oxide synthase and higher expression of tumor necrosis factor-alpha and caspase 3 were found in right ventricle (RV) and lung in group 2 compared with other groups (all p<0.05). The number of alveolar sacs and lung arterioles were also lower in group 2 than in other groups (all p<0.05). Furthermore, RV systolic pressure (RVSP), RV weight, and RV-to-final body weight ratio were substantially increased in group 2 than in other groups, and notably higher in groups 3 and 4 than in groups 1 and 5 (all p<0.0001). CONCLUSIONS: Combined therapy with autologous BMDEPC and sildenafil is superior to either BMDPEC or sildenafil alone for preventing MCT-induced PAH. PMID- 21620491 TI - Aggressive revascularization of acute internal carotid artery occlusion in patients with NIHSS>20 and poor collateral circulation: preliminary report. AB - BACKGROUND: Acute occlusion of internal carotid artery (ICA) is a clinical catastrophic entity with mortality as high as 50%. With innovative devices and technology, we want to clarify the benefit and risk of interventional treatment for those patients. METHODS AND RESULTS: From 2005 to 2009, 62 patients were enrolled and 7 patients were diagnosed as total ICA occlusion with severe neurological deficit and poor collateral circulation received endovascular interventions. Intra-arterial thrombolysis was performed in all the 7 patients. Besides, angioplasty was done in 2 patients, stenting in 3, and thrombosuction in 1. The average NIHSS was 23.3 (standard deviation=3.6) before revascularization, was 14.2(standard deviation=6.8) on day 7. Three patients had symptomatically hemorrhagic transformation and one developed severe brain edema after procedure. Decompressive craniotomy has been conducted in 3, who survived thereafter. One patient died for refusal of decompressive craniotomy. The 30-day modified Rankin scale was 1 in 1, 2 in 1, 3 in 1, and 4 in 3. All of our patients had distal residual lesions at anterior or middle cerebral artery area, and delayed recanalization was noted in 4. CONCLUSIONS: Endovascular therapy was promising as a hyperacute management for patients of ICA total occlusion leading to survival rate more than 80% and significant neurological recovery in 50% of our patients. Distal residual lesions were common in patients of total carotid occlusion after aggressive revascularization. Although the mechanism was not clear, delayed re canalization was common in such patients. PMID- 21620492 TI - Extended surveillance for CBPP in a free country: Challenges and solutions regarding the potential caprine reservoir. AB - Contagious bovine pleuropneumonia (CBPP) is a severe respiratory disease of cattle and buffalo caused by Mycoplasma mycoides subsp. mycoides "Small Colony" (MmmSC). The agent of CBPP has been isolated from goats in different countries including CBPP-free areas. Goats can therefore be regarded as a putative MmmSC reservoir. No diagnostic test for CBPP surveillance in goats has been proposed as yet. Furthermore, serological tests could be seriously hampered by a widespread caprine infection due to the subspecies M. mycoides subsp. capri (Mmc), which is antigenically very close to MmmSC and displays high levels of genetic variability. A competition ELISA (cELISA) is currently used to screen for CBPP in cattle at the herd level in infected areas. The aim of this study was to see if the same cELISA would be specific enough to be used to screen goats despite the potential concomitant infection with Mmc. The cELISA titers of goats from Mmc infected and non-infected herds were comparable and negative using the accepted cutoff for bovine sera. In contrast, seroconversion was observed in goats experimentally inoculated with an Mmc strain that cross-reacted with a monoclonal antibody targeting the same epitope as that used in cELISA. The probability of such false positivity occurring under field conditions is very low since Mmc strains with such an atypical antigenic profile emerge only rarely as a result of random nucleotide variation of the epitope-coding region. In conclusion, the commercially available cELISA can be considered specific enough to be used as a primary test to monitor passage of the CBPP agent in goats, but its sensitivity in goats requires further investigation. PMID- 21620493 TI - Biotechnological implications of the salivary proteome. AB - Although very attractive for noninvasive specimen collection, saliva has not yet been considered a relevant bodily fluid for the diagnosis and prognosis of diseases. The functional roles of specific salivary peptides and proteins have also not yet been studied in detail. Recent proteomic analysis of human whole saliva has shown that salivary biomarkers could contribute to the detection of local and systemic diseases, provided the standardization of proper sampling procedures exists. Recently, interesting and novel functions for different families of specific secretory peptides and proteins have been demonstrated, which could be a basis for the design of peptidomimetics with relevant biotechnological applications. In this review, we focus on the most recent advances in analysing salivary proteins and their potential application in biotechnology. PMID- 21620494 TI - Influence of (11)C-choline PET/CT on the treatment planning for salvage radiation therapy in patients with biochemical recurrence of prostate cancer. AB - BACKGROUND AND PURPOSE: The present study evaluates the incidence of (11)C choline PET/CT positive findings in patients with recurrent prostate cancer referred for salvage radiotherapy (SRT) and the influence on the definition of the planning target volume (PTV). MATERIAL AND METHODS: Thirty-seven patients treated with radical prostatectomy and referred to SRT to the prostatic fossa because of biochemical relapse, were analysed retrospectively. All patients underwent (11)C-choline PET/CT before radiotherapy. The influence of PET/CT on the extent of the PTV was analysed. The median total follow up after SRT was 51.2 months. RESULTS: 11/37 (30%) patients had a positive finding in the (11)C-choline PET/CT, 5 (13%) outside of the prostatic fossa (iliac lymph nodes), implicating an extension of the PTV. Patients with positive (11)C-choline PET/CT had a significant higher PSA value than patients with no pathologic uptake (p=0.03). Overall, at the end of follow up 56% of the patients had a PSA <= 0.2ng/ml and 44% had a biochemical relapse of prostate cancer. CONCLUSIONS: (11)C-choline PET/CT detects abnormalities outside of the prostatic fossa in 13% of patients referred for SRT because of biochemical relapse after radical prostatectomy, affecting the extent of the PTV. Prospective studies need to be implemented to evaluate the benefit of SRT with a PTV based on (11)C-choline PET/CT. PMID- 21620495 TI - Inter-individual and inter-cell type variation in residual DNA damage after in vivo irradiation of human skin. AB - PURPOSE: The aim of this study was to compare inter-individual and inter-cell type variation in DNA double-strand break (DSB) repair following in vivo irradiation of human skin. MATERIALS AND METHODS: Duplicate 4mm core biopsies of irradiated and unirradiated skin were collected from 35 patients 24h after 4Gy exposure using 6MeV electrons. Residual DSB were quantified by scoring 53BP1 foci in dermal fibroblasts, endothelial cells, superficial keratinocytes and basal epidermal cells. RESULTS: Coefficients of inter-individual variation for levels of residual foci 24h after in vivo irradiation of skin were 39.9% in dermal fibroblasts, 44.3% in endothelial cells, 32.9% in superficial keratinocytes and 46.4% in basal epidermal cells (p<0.001, ANOVA). In contrast, the coefficient of inter-cell type variation for residual foci levels was only 11.3% in human skin between the different epidermal and dermal cells (p=0.034, ANOVA). Foci levels between the different skin cell types were correlated (Pearson's R=0.855-0.955, p<0.001). CONCLUSIONS: Patient-specific factors appear to be more important than cell type-specific factors in determining residual foci levels following in vivo irradiation of human skin. PMID- 21620496 TI - Dose escalation in prostate cancer using intensity modulated neutron radiotherapy. AB - BACKGROUND AND PURPOSE: Initial promising results of 3D conformal neutron radiotherapy (3D-CNRT) were subsequently limited by high normal tissue toxicities. It is now possible to deliver intensity modulated neutron radiotherapy (IMNRT). The present work compares photon IMRT, 3D-CNRT and IMNRT for three prostate patients to quantify the benefits of IMNRT. MATERIALS AND METHODS: We compare updated 3D-CNRT plans, IMNRT plans, and conventional IMRT plans by translating neutron DVHs into effective photon DVHs using the dose dependent radiobiological effectiveness (RBE) for each structure. RBE curves are parameterized for a range of normal tissue and prostate tumor values. Generalized equivalent uniform dose (gEUD) and gEUD in 2Gy fractions (gEUD(2)) is calculated for each structure, plan, and parameterization. Rectal sparing and dose to prostate-GTV are compared for 3D-CNRT, IMNRT, and IMRT as a function of normal tissue and prostate RBE. RESULTS: The closer the RBE values of prostate tumor and normal tissue, the greater the advantage of IMNRT over 3D-CNRT. The rectal sparing achieved using IMNRT ranged from ~5% to 13% depending upon the choice of RBE for rectum and the alpha/beta value of prostate tumor. IMNRT may provide a theoretical dose advantage over photon IMRT if the alpha/beta value of prostate is 1.5 and the RBEs of prostate and rectum differ by more than 5%. For higher values of prostate alpha/beta any advantages of IMNRT over IMRT could require that the RBEs of prostate and rectum differ by as much as 20%. CONCLUSIONS: IMNRT provides a clear normal tissue sparing advantage over 3D-CNRT. The advantage increases when the RBEs of the target structure and the normal tissue are similar. This RBE translation method could help identify clinical sites where the dose sparing advantages of IMNRT would allow for the exploitation of the radiobiological advantages of high-LET neutron radiotherapy. PMID- 21620497 TI - Patient-specific PTV margins in radiotherapy for bladder cancer - a feasibility study using cone beam CT. AB - PURPOSE: To assess the feasibility of using cone beam computed tomography (CBCT) to generate patient-specific PTV margins for bladder cancer patients treated with radiation therapy (RT). METHODS: Eleven patients underwent CT simulation and daily RT (full bladder and empty rectum). CBCT was done prior to each fraction, and the whole bladder was contoured off-line. For the first 15 CBCTs of each patient, the bladder was aligned with CT-simulation bladder (pBladder) to create an occupancy volume (OV). A 5mm isotropic margin was added to OV (OV+5). A measurement-based PTV (mPTV) was generated by measuring maximal displacement between pBladder and OV in six directions. OV, OV+5, mPTV, and a standard PTV (2cm isotropic margin) were compared for absolute and relative volume differences. Using the final 10 CBCT of each patient, the ability of each study volume to encompass the entire CBCT bladder was determined. RESULTS: 161/165 CBCT images were of adequate quality for contouring. No daily trend in bladder volume variation was noted. The median absolute volumes (cm(3)) were: 221, 271, 426, 440, and 914 for pBladder, OV, OV+5, mPTV, and standard PTV, respectively. The median ratios of the study volumes/pBladder were: 1.4 OV, 2.1 OV+5, 2.4 mPTV, 4.1 standard PTV. OV+5 was smaller than mPTV in 9 patients. There was considerable inter-patient variability in study volumes and no apparent association of the magnitude of margin expansion and pBladder. The bladder was encompassed in 69%, 99%, 99%, and 100% of the final 10 fractions by OV, OV+5, mPTV, and standard PTV, respectively. CONCLUSIONS: The use of daily CBCT to generate patient-specific PTV margins is feasible and results in a marked reduction in the irradiated volume compared to population-based margins. As daily bladder volume varied unpredictably with considerable differences between patients, these findings support the use of patient-specific PTV margin expansions for bladder radiotherapy. PMID- 21620498 TI - Lung tumor reproducibility with active breath control (ABC) in image-guided radiotherapy based on cone-beam computed tomography with two registration methods. AB - PURPOSE: To study the inter- and intrafraction tumor reproducibility with active breath control (ABC) utilizing cone-beam computed tomography (CBCT), and compare validity of registration with two different regions of interest (ROI). METHODS AND MATERIALS: Thirty-one lung tumors in 19 patients received conventional or stereotactic body radiotherapy with ABC. During each treatment, patients had three CBCT scanned before and after online position correction and after treatment. These CBCT images were aligned to the planning CT using the gray scale registration of tumor and bony registration of the thorax, and tumor position uncertainties were then determined. RESULTS: The interfraction systematic and random translation errors in the left-right (LR), superior-inferior (SI) and anterior-posterior (AP) directions were 3.6, 4.8, and 2.9mm; 2.5, 4.5, and 3.5mm, respectively, with gray scale alignment; 1.9, 4.3, 2.0mm and 2.5, 4.4, 2.9mm, respectively, with bony alignment. The interfraction systematic and random rotation errors with gray scale and bony alignment groups ranged from 1.4 degrees to 3.0 degrees and 0.8 degrees to 2.3 degrees , respectively. The intrafraction systematic and random errors with gray scale registration in LR, SI, AP directions were 0.9, 2.0, 1.8mm and 1.5, 1.7, 2.9mm, respectively, for translation; 1.5 degrees , 0.9 degrees , 1.0 degrees and 1.2 degrees , 2.2 degrees , 1.8 degrees , respectively, for rotation. The translational errors in SI direction with bony alignment were significantly larger than that of gray scale (p<0.05). CONCLUSIONS: With CBCT guided online correction the interfraction positioning errors can be markedly reduced. The intrafraction errors were not diminished by the use of ABC. Rotation errors were not very remarkable both inter and intrafraction. Gray scale alignment of tumor may provide a better registration in SI direction. PMID- 21620499 TI - A population-based study on the utilisation rate of primary radiotherapy for prostate cancer in 4 regions in the Netherlands, 1997-2008. AB - AIM: The purpose was to study variations in utilisation rates of external beam radiotherapy (EBRT) and brachytherapy (BT) for prostate cancer patients. MATERIALS AND METHODS: We calculated the proportion and number of EBRT and BT given or planned within 6 months of diagnosis in 4 Dutch regions, according to stage and age in a population-based setting including 47,259 prostate cancer patients diagnosed from 1997 until 2008. RESULTS: During this study period, the overall utilisation rate of EBRT remained stable at around 25%, while the rate of BT for non-metastasized patients increased from 1% (95% CI:0-1%) to 12% (11-13%) in 2006 and slightly decreased towards 10% (9-11%) in 2008. From 2001 on, the overall utilisation rate of EBRT decreased significantly in one region (p<0.05). In this region, a sharp rise in the utilisation rate of BT for non-metastatic patients was noted to 17% (14-20%) in 2008 after a peak of 24% (21-27%) in 2006. For localised disease, BT was used more often at the expense of EBRT while for locally advanced disease the utilisation rate of EBRT increased. In the multivariate analysis, regional differences in the utilisation rate of EBRT persisted with odds ratios ranging from 0.7 to 0.9 compared to the reference region. Moreover, low rates of EBRT were associated with high BT rates. The regional differences could not be explained by differences in risk profiles. CONCLUSIONS: The utilisation rate of EBRT remained stable with limited variation between regions while BT was used increasingly with clear regional differences. To cope with this and in view of the increasing incidence of prostate cancer, adequate resources have to be planned for the optimal care of these patients. PMID- 21620500 TI - Association between single nucleotide polymorphisms in the DNA repair gene LIG3 and acute adverse skin reactions following radiotherapy. AB - Many genes have been associated with radiotherapy toxicity, but most have only been found in a single study. Using our cohort of 480 breast cancer patients, we provide replicated evidence that a polymorphism near the LIG3 gene is associated with acute skin toxicity following radiotherapy. PMID- 21620501 TI - Displacement of hepatic tumor at time to exposure in end-expiratory-triggered pulse proton therapy. AB - PURPOSE: This study is to evaluate reproducibility of hepatic tumors in end expiration and end-inspiration on free-breathing, also measure shift of hepatic tumor location in pulsed proton beams exposure in end-expiration in order to estimate feasible planning target volume (PTV) margin. MATERIALS AND METHODS: Pairs (1232) of anterior and lateral radiographs from 30 patients (628 end expiration and 604 end-inspiration phases) were analyzed using fiducial markers adjacent to the tumors. By using the co-ordinates of the marker centroid of mass related to the isocenter, intrafractional variation was compared in end expiration and end-inspiration, and a feasible PTV margin was generated using the measured motion. RESULTS: The average internal motion in end-expiration was 1.1mm, which was significantly smaller than that in end-inspiration. The mean deviation from the plan was -0.1, 0.3, and 0.1mm in the left-right (LR), cranio cepharal (CC), and anterior-posterior (AP) directions, respectively. The estimated PTV margins were 3.2, 3.5, and 4.6mm, in the LR, CC, and AP directions, respectively. CONCLUSIONS: It was indicated that localization of the targets was more reproducibility in end-expiration than that in end-inspiration. Also, feasible and practical margin values were obtained. These should contribute accuracy of respiration synchronized proton radiotherapy for liver tumors. PMID- 21620502 TI - Whole brain radiotherapy: prognostic factors and results of a radiation boost delivered through a conventional linear accelerator. AB - PURPOSE: Assess prognostic factors for overall survival and the potential benefit of a boost in patients treated with whole brain radiation therapy (WBRT). METHODS AND MATERIALS: From 2002 to 2006, a retrospective analysis was made from 250 unselected consecutive patients with secondary brain metastases from lung cancer, breast cancer and melanoma. Eighteen patients received surgery and were excluded from analysis. Four potential prognostic factors have been studied: primary tumor type, gender, number of metastases and improvement of neurological symptoms after radiation therapy. A subgroup analysis was performed to determine whether an additional boost could potentially improve outcome in patients who presented with less than three metastases, performance status <2, and no surgical resection of their metastasis. RESULTS: Average follow-up was 10.3 months. Median overall survival was 5.6 months and survival rates at 1 and 2 years were 22.7% and 10%, respectively. Age less than 65 (p<0.01), neurological improvement after WBRT (p<0.01), and presence of less than three metastases were significant factors for overall survival in multivariate analysis. When focusing on the selected subgroup (120 assessable patients), median overall survival was 4.0 months in patients with no radiation boost, versus 8.9 months in patients with radiation boost (p=0.0024). CONCLUSIONS: Survival and prognostic factors were similar to those found in the literature. Boost delivered after WBRT by a conventional particle accelerator could provide a benefit in selected patients, especially for centers that do not have radiotherapy techniques in stereotactic conditions. This warrants further prospective assessment. PMID- 21620503 TI - Withholding stereotactic radiotherapy in elderly patients with stage I non-small cell lung cancer and co-existing COPD is not justified: outcomes of a Markov model analysis. AB - BACKGROUND AND PURPOSE: To model outcomes of SBRT versus best supportive care (BSC) in elderly COPD patients with stage I NSCLC. MATERIAL AND METHODS: A Markov model was constructed to simulate the quality-adjusted and overall survival (OS) in patients ?75years undergoing either SBRT or BSC for a five-year timeframe. SBRT rates of local, regional and distant recurrences were obtained from 247 patients treated at the VUMC, Amsterdam. Recurrence rates were converted into transition probabilities and stratified into four groups according to T stage (1, 2) and COPD GOLD score (I-II, III-IV). Data for untreated patients were obtained from the California Cancer Registry. Tumor stage and GOLD score utilities were adapted from the literature. RESULTS: Our model correlated closely with the source OS data for SBRT treated and untreated patients. After SBRT, our model predicted for 6.8-47.2% five-year OS and 14.9-27.4 quality adjusted life months (QALMs). The model predicted for 9.0% and 2.8% five-year OS, and 10.1 and 6.1 QALMs for untreated T1 and T2 patients, respectively. The benefit of SBRT was the least for T2, GOLD III-IV patients. CONCLUSION: Our model indicates that SBRT should be considered in elderly stage I NSCLC patients with COPD. PMID- 21620504 TI - Cervical nodal metastases of unresectable thoracic esophageal squamous cell carcinoma: characteristics of long-term survivors after concurrent chemoradiotherapy. AB - PURPOSE: To analyze the clinical implications of metastatic cervical lymph nodes in unresectable thoracic esophageal squamous cell carcinoma (SCC) after concurrent chemoradiotherapy (CRT). METHODS AND MATERIALS: 208 thoracic esophageal SCC patients treated with concurrent CRT were analyzed retrospectively. Patients were divided into 3 groups according to different status of metastatic cervical lymph nodes: 1. CLN(-), no evidence of metastatic cervical lymph nodes; 2. CPLN(+), evidence of enlarged cervical paraesophageal lymph node without any other metastatic cervical lymph nodes; 3. OCLN(+), any other situations of enlarged cervical lymph nodes. The prognostic factors were examined univariately, then selected for inclusion in a multivariate Cox regression model. RESULTS: Three-year OS of CLN(-),CPLN(+), and OCLN(+) groups were 39%, 33%, and 15% (logrank p=0.001). On univariate analysis, variables significantly associated with OS included sex, primary esophageal tumor location, length and cervical nodal disease. On multivariate analysis, primary tumor location (HR1.5, 95%CI 1.1-1.9, p=0.005), tumor length (HR1.5, 95%CI 1.1-2.0, p=0.010) and cervical nodal disease (HR1.2, 95%CI 1.0-1.5, p=0.014) were prognostic factors on OS. CONCLUSIONS: (1) Primary tumor location, tumor length, and cervical nodal disease were significant prognostic factors on OS in esophageal SCC patients. (2) Patients with CPLN(+) disease tended to have a potential better long-term survival than those with OCLN(+) disease, and metastatic CPLN could be considered as local disease. The survival benefit in CPLN(+) subgroup might be contributed by the patients who presented with upper third thoracic tumors and shorter tumor length. Further investigation is needed. PMID- 21620505 TI - New insights into physiology and metabolism of Propionibacterium freudenreichii. AB - Dairy propionibacteria are Actinobacteria, mainly isolated from dairy environments. Propionibacterium freudenreichii has been used for a long time as a ripening culture in Swiss-type cheese manufacture, and is more and more considered for its potent probiotic effects. This review summarises the knowledge on the main P. freudenreichii pathways and the main features explaining its hardiness, and focuses on recent advances concerning its applications as a cheese ripening agent and as a probiotic for human health. Propionibacteria have a peculiar metabolism, characterised by the formation of propionic acid as main fermentation end-product. They have few nutritional requirements and are able to use a variety of carbon substrates. From the sequence of P. freudenreichii CIRM BIA1(T) genome, many pathways were reconstituted, including the Wood-Werkman cycle, enzymes of the respiratory chain, synthesis pathways for all amino acids and many vitamins including vitamin B(12). P. freudenreichii displays features allowing its long-term survival. It accumulates inorganic polyphosphate (polyP) as energy reserve, carbon storage compounds (glycogen), and compatible solutes such as trehalose. In cheese, P. freudenreichii plays an essential role in the production of a variety of flavour compounds, including not only propionic acid, but also free fatty acids released via lipolysis of milk glycerides and methyl butanoic acids resulting from amino acid degradation. P. freudenreichii can exert health-promoting activities, such as a bifidogenic effect in the human gut and promising immunomodulatory effects. Many P. freudenreichii properties involved in adaptation, cheese ripening, bio-preservation and probiotic effects are highly strain-dependent. The elucidation of the molecular mechanisms involved is now facilitated by the availability of genome sequence and molecular tools. It will help in the selection of the most appropriate strain for each application. PMID- 21620506 TI - Influence of different floor management strategies of the vineyard on the natural yeast population associated with grape berries. AB - Some oenological practices, such as the massive utilisation of commercial yeast and the consequent colonisation of wineries, can contribute to reducing the native yeast biodiversity. In this context, the vineyard could be a reservoir of autochthonous yeasts of oenological interest. Thus, the evaluation of the influence of different agricultural parameters on the biodiversity of yeast population in the vineyard is necessary. This work shows the results of the influence of some floor management strategies of the vineyard in the natural yeast population associated with the grape-berries. With this objective, a three year sampling plan was designed in the Shiraz vineyards of the Madrid region using three floor management strategies: bare soil by tillage, bare soil maintained with herbicides and soil maintained with cover crop. The results of this study have shown that bare soil by tillage could be a sustainable alternative for managing the soil, due to the reduced use of agrochemicals and the resulting high yeasts biodiversity. Nevertheless, the presence of herbicides in the vineyard has a minor impact on the diversity of grape associated yeast communities, and this could have increased the yeast populations. Hence, from the fermentative yeasts' (like Saccharomyces) point of view, in hot and arid environments where soils may be affected by the tillage management, the best option could be the maintenance of the bare soil with the use of herbicides. PMID- 21620507 TI - Prevalence and mean intensity of Anisakis simplex (sensu stricto) in European sea bass (Dicentrarchus labrax) from Northeast Atlantic Ocean. AB - Viscera and muscle of a total of 40 wild 1-2kg European sea bass (Dicentrarchus labrax) from Northeast Atlantic (FAO area 27) were examined for Anisakidae larvae detection by digestion method. Extracted parasites were counted and mean intensity was calculated. Parasites were identified by genetic/molecular markers (allozymes and sequences analysis of the mtDNA cox2 gene) as belonging to the species Anisakis simplex (sensu stricto). In viscera, the main localisations of the larvae were under the gastric serosa, where several parasites alive and dead were found, and intestinal serosa. The visceral prevalence was 0.950 and the mean intensity was 96.39. The main localisation of A. simplex (s.s.) in edible parts was in belly muscles, with a prevalence of 0.425 and a mean intensity of 1.9. This is the first record on the prevalence and mean intensity of A. simplex (s.s.) in European sea bass muscle. This finding has an important consequence on epidemiology of anisakiasis and public health risk assessment. PMID- 21620508 TI - Epidemiology of spotted wilt disease of peanut caused by Tomato spotted wilt virus in the southeastern U.S. AB - Spotted wilt disease of peanut (Arachis hypogaea) (SWP), caused by Tomato spotted wilt virus (TSWV) (genus Tospovirus, family Bunyaviridae), was first observed in Alabama, Florida, and Georgia in the late 1980s and rapidly became a major limiting factor for peanut production in the region. Tobacco thrips (Frankliniella fusca) and western flower thrips (Frankliniella occidentalis) both occur on peanut throughout the southeastern U.S., but F. fusca is the predominant species that reproduces on peanut, and is considered to be the more important vector. Several non-crop sources of potential primary vectors and TSWV inoculum have been identified, but their relative importance has not been determined. The peanut growing season in Alabama, Florida, and Georgia is from April through November, and 'volunteer' peanut plants can be present for much of the remainder of the year. Therefore peanut itself has huge potential for perpetuating both vector and virus. Symptoms are often evident within a few days of seedling emergence, and disease progress is often rapid within the first 50-60 days after planting. Based on destructive sampling and assays for TSWV, there is often a high incidence of asymptomatic infections even in peanut genotypes that produce few and mild symptoms of infection in the field. Severity of SWP epidemics fluctuates significantly from year to year. The variability has not been fully explained, but lower incidences have been associated with years categorized as "La Nina" in the El Nino-Southern Oscillation. Planting date can have a large effect on disease incidence within a location. This may be linked to the thrips reproductive cycle and environmental effects on the plant and plant-thrips-virus interactions. Row pattern, plant population, and in-furrow applications of phorate insecticide can also affect epidemics of SWP. Considerable progress has been made in developing cultivars with natural field resistance to TSWV. Use of cultivars with moderate field resistance combined with other suppressive measures has been very successful for managing spotted wilt disease. Several new cultivars with higher levels of field resistance can improve control and allow more flexibility in the integrated management programme. Although effects of these factors on epidemics of SWP have been documented, mechanisms responsible for disease suppression by most factors have not been fully elucidated. PMID- 21620509 TI - Application of Australian clinical management guidelines: the current state of play in a sample of young people living with Type 1 diabetes in the state of New South Wales and the Australian Capital Territory. AB - AIMS: To describe care provided to a sample of young Australians with Type 1 diabetes, and benchmark this against national guidelines. METHODS: 158 children and adolescents with Type 1 diabetes, aged 8-19 years, were recruited independent of their source of care as part of a three-year longitudinal study. Data were gathered annually regarding type of health-care services attended, demographic, health-care and self-care information. Participants were also telephoned quarterly to ascertain planned and actual attendance to diabetes services, and current diabetes management. A capillary sample was collected annually for HbA1c determination. RESULTS: The mean HbA1c of participants was significantly higher than recommended levels. The annual number of visits to diabetes clinics also fell short of the stipulated 3-4 visits a year and less than 25% of participants received care from all recommended multidisciplinary team members. While the majority of care was provided through the publicly funded system, there was an increasing reliance on privately funded psychologists. CONCLUSION: Standards of care received by this group of young Australians and levels of glycaemic control fall short of treatment guidelines, highlighting the need to identify ways to ensure equitable access to specialist multidisciplinary care for all young people affected by diabetes. PMID- 21620510 TI - Cigarette smoking and lung cancer in women: results of the French ICARE case control study. AB - BACKGROUND: The incidence of female lung cancer in developed countries has been increasing since 1950. In order to have recent and reliable data on the association between cigarette smoking and the risk of lung cancer in women, we analysed cases from a French population-based case-control study. METHODS: The ICARE study is a multicenter case-control study on respiratory cancers (lung and UADT cancers), set up in 10 departements that include a general cancer registry. We included 648 women lung cancer cases up to 76 years of age, with a histologically confirmed primary lung cancer. The 775 controls were randomly selected from the general population and frequency-matched with cases by age and departement. RESULTS: Overall, smoking cigarettes at some time was associated with a 8-fold increase in lung cancer risk (OR=8.2, 95% CI 6.0-11.4). A dose response relationship was observed as a function of duration, intensity and pack years. Using restricted splines cubic models, we have shown that intensity dose response departed significantly from linearity while the risk increased linearly with duration and decreased linearly with time since cessation. The following characteristics were associated with a higher relative risk: smoke inhalation, smoking non-filter cigarettes, smoking dark tobacco cigarettes and starting at a young age. In addition, duration, intensity and time since cessation was significantly related with histological type. This was not the case for characteristics such as the use of a filter or not, the inhalation pattern, or the type of tobacco smoked. The proportion of lung cancer cases attributable to cigarette smoking was 55% (95% CI: [47-63%]). CONCLUSIONS: Our results confirm that cigarette smoking is by far the most important cause of the current epidemic of lung cancer among French women and that the most important smoking-related variables for varying the risk of lung cancer are the duration, the intensity and the time since cessation. PMID- 21620511 TI - A phase II study of obatoclax mesylate, a Bcl-2 antagonist, plus topotecan in relapsed small cell lung cancer. AB - INTRODUCTION: We previously reported data on the safety, tolerability, and recommended phase II dose of obatoclax mesylate in conjunction with topotecan in patients with advanced solid tumor malignancies. Preliminary efficacy data suggested activity in patients with recurrent small cell lung cancer (SCLC). Based on these data, we performed a phase II study of obatoclax mesylate plus topotecan in patients with relapsed SCLC to assess efficacy. METHODS: This was an open-label, single-arm, phase II extension of obatoclax mesylate plus topotecan in patients with relapsed SCLC. Obatoclax mesylate was given intravenously (IV) at a dose of 14mg/m(2) on days 1 and 3 with IV topotecan at 1.25mg/m(2) on days 1 5 of an every 3-week cycle. The primary end-point of this study was overall response rate. RESULTS: Nine patients with recurrent SCLC were enrolled into the first stage of the study. Patients received a median of 2 cycles of treatment. All patients were evaluable for the primary end-point of overall response. There were no partial or complete responses. Five patients (56%) had stable disease. The remaining four patients (44%) developed progressive disease. The most common grade 3 or 4 adverse events included thrombocytopenia (22%), anemia (11%), neutropenia (11%), and ataxia (11%). CONCLUSION: Obatoclax mesylate added to topotecan does not exceed the historic response rate seen with topotecan alone in patients with relapsed SCLC following the first-line platinum-based therapy. PMID- 21620513 TI - Group selection and kin selection: formally equivalent approaches. AB - Inclusive fitness theory, summarised in Hamilton's rule, is a dominant explanation for the evolution of social behaviour. A parallel thread of evolutionary theory holds that selection between groups is also a candidate explanation for social evolution. The mathematical equivalence of these two approaches has long been known. Several recent papers, however, have objected that inclusive fitness theory is unable to deal with strong selection or with non additive fitness effects, and concluded that the group selection framework is more general, or even that the two are not equivalent after all. Yet, these same problems have already been identified and resolved in the literature. Here, I survey these contemporary objections, and examine them in the light of current understanding of inclusive fitness theory. PMID- 21620512 TI - Chemotherapy and targeted therapies for unresectable malignant mesothelioma. AB - The global burden of mesothelioma is expected to increase in the coming decades. As a result the development of more effective therapies with an emphasis on personalized treatments based on validated prognostic and predictive biomarkers is an essential requirement. Progress has been made in the last decade with the development of newer generation anti-folates leading to the current standard of care of pemetrexed and cisplatin in patients with unresectable disease. However, the median overall survival of patients with this combination treatment is only 12 months. There is no consensus regarding second line therapy for patients who have progressed or not responded to pemetrexed based therapies although gemcitabine in combination with a platinum compound or single agent vinorelbine is a reasonable option. The development of effective targeted agents that are active in mesothelioma has to date been disappointing. Strategies involving the addition of bevacizumab to pemetrexed and cisplatin in the frontline setting, the histone deacetylase inhibitor vorinostat as second line therapy and studies evaluating the utility of maintenance therapy in mesothelioma are all ongoing and appear promising. In addition clinical trials investigating immunotherapy and gene therapy in combination with chemotherapy could potentially improve the prognosis of patients with mesothelioma. PMID- 21620514 TI - Convergent or parallel molecular evolution of momilactone A and B: potent allelochemicals, momilactones have been found only in rice and the moss Hypnum plumaeforme. AB - Plant second metabolites momilactone A and B, which act as potent phytoalexins and allelochemicals, have been found thus far only in rice and the moss Hypnum plumaeforme, although both plants are taxonomically quite distinct. The concentrations of momilactone A and B, respectively, in rice plants were 4.5-140 and 2.9-85MUg/g, and those in H. plumaeforme were 8.4-58.7 and 4.2-23.4MUg/g. Momilactone A and B concentrations in rice and H. plumaeforme plants were increased by UV irradiation, elicitor and jasmonic acid treatments. Rice and H. plumaeforme plants secrete momilactone A and B into the rhizosphere, and the secretion level was also increased by UV irradiation, elicitor and jasmonic acid treatments. In addition, although endogenous concentrations of momilactone A in rice and H. plumaeforme were greater than those of momilactone B, the secretion levels of momilactone B were greater than those of momilactone A in rice and H. plumaeforme, which suggests that momilactone B may be selectively secreted by both rice and H. plumaeforme. As momilactone A and B exert potent antifungal and growth inhibitory activities, momilactone A and B may play an important role in the defense responses in H. plumaeforme and rice against pathogen infections and in allelopathy. The secretion of momilactone A and B into the rhizosphere may also prevent bacterial and fungal infections and provide a competitive advantage for nutrients through the inhibition of invading root systems of neighboring plants as allelochemicals. Therefore, both plants, despite their evolutionary distance, may use same defense strategy with respect to the momilactone A and B production and secretion, which resulting from convergent or parallel evolutionary processes. In the case of parallel evolution, there may be plant species providing the missing link in molecular evolution of momilactones between H. plumaeforme and rice. PMID- 21620515 TI - A randomized, investigator-blinded, controlled pilot study to evaluate the safety and efficacy of a poly-N-acetyl glucosamine-derived membrane material in patients with venous leg ulcers. AB - BACKGROUND: Standard care for venous leg ulcers (VLUs) has remained unchanged over several decades despite high rates of initial treatment failure and ulcer recurrence. OBJECTIVE: We sought to evaluate the efficacy, safety, and tolerability of an advanced, poly-N-acetyl glucosamine (pGlcNAc), nanofiber derived, wound-healing technology among patients with VLUs (Talymed, Marine Polymer Technologies Inc, Danvers, MA). METHODS: In this randomized, investigator blinded, parallel-group, controlled study, eligible patients were randomized to treatment with standard care plus pGlcNAc (applied only once, every other week, or every 3 weeks) or to standard care alone. The primary end point was the proportion of patients with complete wound healing at week 20 in the intent-to treat population (all randomized subjects), with last observation carried forward. RESULTS: Among 82 randomized patients, 71 completed the study with 7 lost to follow-up and 4 discontinued because of systemic infection. There were no significant group differences with regard to baseline demographic, illness, and VLU characteristics. At 20 weeks, the proportion of patients with completely healed VLUs was 45.0% (n = 9 of 20), 86.4% (n = 19 of 22), and 65.0% (n = 13 of 20) for groups receiving standard care plus pGlcNAc only once, every other week, and every 3 weeks, respectively, versus 45.0% (n = 9 of 20) for those receiving standard care alone (P < .01 for pGlcNAc every other week vs standard care). The novel pGlcNAc advanced wound-healing technology was well tolerated and safe. LIMITATIONS: Limitations were small sample size and patients unblinded to treatment allocation. CONCLUSION: These pilot study results suggest that the pGlcNAc advanced wound-healing technology is well tolerated and effective. PMID- 21620516 TI - Narrowband ultraviolet B phototherapy is a suitable treatment option for solar urticaria. AB - BACKGROUND: Narrowband (NB) ultraviolet (UV) B lamps are widely used for treatment and prophylaxis of several skin diseases. OBJECTIVE: We sought to assess the efficacy of two protocols of NB-UVB therapy for the prophylaxis of UVB sensitive and UVB-insensitive solar urticaria (SU). METHODS: Subjects affected by SU underwent phototesting for assessment of the minimal erythemal dose and minimal urticarial dose. Patients without urticarial response to UVB underwent a single daily exposure every other day for 4 weeks (group A). Patients with a urticarial test response to broadband UVB or NB-UVB (group B) received 3 daily exposures (on working days) for the first week. Afterward, they were treated as the patients of group A for 3 weeks. Follow-up visits took place after 1 and 3 months. RESULTS: A total of 39 patients completed the study. In groups A (29 patients) and B (10 patients), the median total number of exposures was 12 (interquartile range [IQR]: 12; 15) and 25.5 (IQR 24; 27), respectively. The median total NB-UVB dose was 10.3 J/cm(2) (IQR 9.9; 11) for group A and 9.1 J/cm(2) (IQR 8.5; 10.6) for group B. At follow-up visits, patients reported good tolerance to the sun. LIMITATIONS: A direct comparison of NB-UVB with UVA or psoralen plus UVA for the photoprophylaxis of SU is still lacking. CONCLUSION: NB UVB phototherapy was well-tolerated and effectively prevented SU relapses. PMID- 21620517 TI - In vivo reflectance confocal microscopy of extramammary Paget disease: diagnostic evaluation and surgical management. AB - BACKGROUND: Extramammary Paget disease (EMPD) is a diagnostic challenge. In vivo reflectance confocal microscopy (RCM) has been reported to be useful for in vivo skin tumor evaluation. It may also assist in the surgical management of EMPD lesions. OBJECTIVE: We sought to describe confocal features of EMPD and correlate them with histopathologic findings. The potential of RCM to map the lesions for subsequent surgical management was also investigated. METHODS: A total of 23 lesions from 14 recruited patients were evaluated by RCM and histopathologic examination. RCM was used to delineate preoperative surgical margins in two patients. RESULTS: Erythematous, hyperpigmented, and hypopigmented lesions were evaluated by RCM and results were confirmed by histopathologic examination. Paget cells were observed throughout the epidermis. Typical Paget cells on RCM were characterized by a mild bright nucleus and dark cytoplasm, frequently twice the size of keratinocytes or larger. At the dermoepidermal junction, tumor nests were seen as dark glandular structures. A high density of dendritic cells was observed in pigmented lesions and a low density in erythematous lesions. Dilated vessels and inflammatory cells were seen in pigmented and erythematous lesions. Paget cells within the epidermis and nest structures at the dermoepidermal junction were seen in most lesions. These two features were useful for delineating the margins. Histologic examination corroborated the surgical margins found by RCM. LIMITATIONS: The sensitivity and specificity of these diagnostic features have not been fully studied, and differential diagnostic features require exploration. CONCLUSION: Features correlating well to histopathology are observed on the RCM of EMPD lesions. RCM may be used as an auxiliary diagnostic tool for the diagnosis and management of EMPD. PMID- 21620518 TI - Neuropeptide Y expression in cutaneous melanoma. AB - BACKGROUND: Neuropeptide Y (NPY) is widely found in the nervous system and has a role in numerous physiologic processes. In addition, NPY receptors are expressed in neuroendocrine tumors, breast cancer, prostate cancer, kidney cancer, and some types of sarcomas. Different neuropeptides, particularly alpha-melanocyte stimulating hormone (MSH), seem to play a role in the pathogenesis of melanoma. OBJECTIVE: We sought to analyze the expression of NPY in cutaneous melanoma, its association with clinical and histologic features, and its correlation with alpha MSH. METHODS: This was an observational study of the immunohistochemical expression of NPY and alpha-MSH in tissue samples of cutaneous melanomas, different types of melanocytic nevi, and melanoma metastases diagnosed from 2004 to 2008 in San Jorge Hospital, Huesca, Spain. RESULTS: A total of 184 lesions were studied: 49 primary cutaneous melanomas, 12 melanoma metastases (9 cutaneous and 3 lymphatic), and 123 melanocytic nevi. Immunostaining revealed that levels of NPY and alpha-MSH were significantly higher in melanomas than in melanocytic nevi (P < .001). Melanoma metastases were negative for both neuropeptides. Nodular melanomas showed the highest median percentage of NPY positive cells (75% [20-95]) followed by superficial spreading melanoma (25% [2-92]), whereas lentigo maligna were negative (0% [0-0]). Significant, direct associations between NPY expression and vertical growth (P = .0141) and presence of metastasis (P = .0196) were observed. NPY and alpha-MSH were positively correlated in cutaneous melanoma (0.49, P < .001). LIMITATIONS: The sample size of melanomas was not very large. CONCLUSION: Our study demonstrates that NPY is significantly expressed in melanomas, especially the nodular type, being associated with invasiveness independently of proliferative markers such as thickness, ulceration, and mitotic index. PMID- 21620519 TI - Preventing methicillin-resistant Staphylococcus aureus transmission in hospitals: an Executive Summary of the Association for Professionals in Infection Control and Epidemiology, Inc, Elimination Guide. AB - This article is an executive summary of the Association for Professionals in Infection Control and Epidemiology, Inc, Elimination Guide for methicillin resistant Staphylococcus aureus, including the 2009 California Supplement. Infection preventionists are encouraged to obtain the original, full-length Association for Professionals in Infection Control and Epidemiology, Inc, Elimination Guides for more thorough coverage of Staphylococcus aureus prevention. PMID- 21620520 TI - Outbreak or coincidental cases of tuberculosis? Genotyping provides the clue. AB - We illustrate how genotyping of mycobacterial strains contributed to the discovery of an undetected outbreak of tuberculosis in a hospital ward, ruling out misleading assumptions of transmission chains. Genotyping should be taken into account in routine tests for the control of tuberculosis. PMID- 21620521 TI - [Laparoscopic partial adrenalectomy for aldosterone-producing adenoma]. PMID- 21620522 TI - [Comment to "Association of nocturnal penile rigidity with testosterone, metabolic syndrome, and other variables: a prospective cross-sectional pilot study"]. PMID- 21620523 TI - [Assessment of cardiovascular preload and response to volume expansion]. AB - Volume expansion is used in patients with hemodynamic insufficiency in an attempt to improve cardiac output. Finding criteria to predict fluid responsiveness would be helpful to guide resuscitation and to avoid excessive volume effects. Static and dynamic indicators have been described to predict fluid responsiveness under certain conditions. In this review we define preload and preload-responsiveness concepts. A description is made of the characteristics of each indicator in patients subjected to mechanical ventilation or with spontaneous breathing. PMID- 21620524 TI - [Prolonged stay in pediatric intensive care units: mortality and healthcare resource consumption]. AB - OBJECTIVE: To analyze mortality and resource consumption in patients with long stays in pediatric intensive care units (PICUs). DESIGN: A retrospective, descriptive case series study. SCOPE: Medical-surgical PICU in a third level hospital. PATIENTS: Data were collected from patients with a stay of 28 days or more in PICU between 2006 and 2010. Of the 2118 patients assisted in this period, 83 (3.9%) required prolonged stay. STUDY VARIABLES: Morbidity-mortality and resource consumption among patients with prolonged stay in the PICU. RESULTS: Mortality was higher in patients with a long stay (22.9%) than in the rest of patients (2%) (p<0.001). In 52.6% of these patients, death occurred after withdrawal of treatment or after not starting resuscitation measures. Patients with prolonged stay showed a high incidence of nosocomial infection (96.3%) and an important consumption of healthcare resources (97.6% required conventional mechanical ventilation, 90.2% required transfusion of blood products, 86.7% required intravenous vasoactive drugs and 22.9% required extracorporeal membrane oxygenation [ECMO]). CONCLUSIONS: Critical children with prolonged stay in the PICU show important morbidity and mortality, and an important consumption of healthcare resources. The adoption of specific measures permitting early identification of patients at risk of prolonged stay is needed in order to adapt therapeutic measures and available resources, and to improve treatment efficiency. PMID- 21620525 TI - [Iliopsoas muscle abscess]. PMID- 21620526 TI - [Determination of the prevalence of restless legs syndrome in the adult population of Burgos]. PMID- 21620528 TI - [Evaluation of chromID ESBL medium for detecting carriers of extended-spectrum beta-lactamase-producing enterobacteriaceae]. PMID- 21620527 TI - [Resistance to first line chemotherapy treatment in a patient diagnosed of nodular lymphocyte-predominant Hodgkin's lymphoma identified by 18F-FDG PET/CT: diagnostic and therapeutic implications]. AB - Nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL) is a rare entity that accounts for less than 5% of the cases of Hodgkin lymphoma (HL) with morphological, immunophenotypical, genetic and clinical behavior traits different from the classic HL. In a minority of patients, the NLPHL course is complicated by a transformation to a non-Hodgkin diffuse large B-cell lymphoma (NHDLBCL) with prognostic and therapeutic implications. Early metabolic changes observed by (18)F-FDG PET in patients with HL and NHL, after 1-3 cycles of chemotherapy, predict the final response to treatment and progression-free survival. In the case we are presenting herein, whether NLPHL is transformed to NHDLBCL or the two types of lymphoma co-exist in the same patient, the (18)F-FDG PET/CT scan was crucial for the identification of tumor resistance to first line chemotherapy and to guide a second biopsy decision and therefore modify the chemotherapy regimen. PMID- 21620529 TI - A class of Trp-Trp-AA-OBzl: Synthesis, in vitro anti-proliferation/in vivo anti tumor evaluation, intercalation-mechanism investigation and 3D QSAR analysis. AB - From the anti-tumor active N-tryptophanyl-beta-carboline-3-carboxylic acid benzyl ester and beta-carboline-3-carbonyltryptophan benzyl ester, a pharmacophore, Trp Trp-OBzl, was drawn. Based on the DOCK scores amino acid residue was inserted into the C-terminus of Trp-Trp-OBzl and twenty Trp-Trp-AA-OBzls (AA = amino acid residues) were provided as DNA intercalators. On the in vitro and in vivo models seventeen Trp-Trp-AA-OBzls were anti-tumor active, and twelve Trp-Trp-AA-OBzls were more active than cytarabine. In acute toxicity assay Trp-Trp-AA-OBzls did not damage the immunologic function and had an LD(50) of more than 500 mg/kg. The relationships of structure and activity were analyzed with 3D QSAR. The action mechanism studies revealed that the in vivo anti-tumor action of Trp-Trp-AA-OBzls was the result of DNA intercalation. PMID- 21620530 TI - Semi-synthesis and antitumor activity of 6-isomers of 5, 8-O-dimethyl acylshikonin derivatives. AB - We recently discovered that 5, 8-O-dimethyl acylshikonin derivatives displayed the selectivity towards MCF-7 and no toxicity to normal cells. Herein, a series of the corresponding 6-isomers of 5, 8-O-dimethyl acylshikonin derivatives were synthesized starting from shikonin. In vitro evidence of the cytotoxicities indicated that most of thecompounds were more active than or comparative to shikonin and retained the selectivity against MCF-7, MDA-MB-231 besides no toxicity in the normal cells. Also, in vivo anticancer activity of the positional isomers 5p, 6c further showed that 6-isomers of 5, 8-O-dimethyl acylshikonin derivatives were more active than their corresponding 2-isomers. Thus, we may conclude that the position of the side chain of shikonin attached to 5,8 dimethoxy -1,4-naphthoquinone is associated with the antitumor activity. PMID- 21620531 TI - Synthesis and biological evaluation of conformationally flexible as well as restricted dimers of monastrol and related dihydropyrimidones. AB - A series of conformationally flexible and restricted dimers of monastrol as well as related dihydropyrimidones have been synthesized by employing one-pot Biginelli multicomponent reaction. These dimers have been evaluated for cytotoxic potency against selected human cancer cell lines and some of the compounds have exhibited more cytotoxic potency than the parent monastrol. Further, the DNA binding ability by thermal denaturation studies and antimicrobial activities of these compounds are also discussed. PMID- 21620532 TI - Conjugates of 3alpha-methoxyserrat-14-en-21beta-ol (PJ-1) and 3beta-methoxyserrat 14-en-21beta-ol (PJ-2) as cancer chemopreventive agents. AB - 3alpha-Methoxyserrat-14-en-21beta-ol (PJ-1) and 3beta-methoxyserrat-14-en-21beta ol (PJ-2) were conjugated with well-known phenolic compounds, narigenin, hesperetin, genistein, and daidzein (1-8). Other conjugates of PJ-2-3,5-dihydroxy 4-methoxybenzoic acid (9), PJ-2-pyrogallol (10), and derivatives of PJ-1, PJ-2 3,3-dimethyl-succinates (11, 12), PJ-1, PJ-2-succinates (13, 14), PJ-2-glycine (15), PJ-2-piperidine acetic acid (16), and PJ-1 epoxy-3,3-dimethyl-succinate (17) were tested for their inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by 12-O-tetradecanoylphorbol-13-acetate (TPA). The inhibitory effects of 11 (IC(50) = 251), 12 (IC(50) = 248), and 17 (IC(50) = 230 mol ratio/32 pmol/TPA), were 2-fold stronger than those of the other compounds such as oleanolic acid (IC(50) = 449). Compounds 10, 11, and 17 inhibited mouse skin tumor promotion in an in vivo two-stage carcinogenesis model. The in vivo two-stage mouse-skin carcinogenesis test employed 7,12-dimethylbenz[a]anthracene (DMBA) as an initiator and TPA as a promoter. PMID- 21620533 TI - Synthesis and biological evaluation of potential 5-HT(7) receptor PET radiotracers. AB - Brain serotonin 7 receptor (5-HT(7)) is involved in several mood disorders and drug candidates targeting this subtype are currently in development. Positron emission tomography (PET) is a molecular imaging modality offering great promise for accelerating the process from preclinical discovery to clinical phases. As no PET radiopharmaceutical has yet been used successfully to study the 5-HT(7) receptor in vivo, our objective is to develop the first 5-HT(7) fluorine-18 labeled radiotracer. Four structural analogs of SB269970, a specific 5-HT(7) receptor antagonist, divided in FP3 series and FPMP series were synthesized. Their antagonist effects were investigated by cellular functional assay. Nitro precursors of these analogs were radiolabeled via a [(18)F(-)]nucleophilic substitution and in vitro autoradiographies were performed in rat brain. Chemical and radiochemical purities of fluorine radiotracers were >99% with specific activities in 40-129 GBq/MUmole range. The four derivates presented antagonism potencies toward 5-HT(7) receptors (pK(B)) between 7.8 and 8.8. The four PET radiotracers had suitable characteristic for 5-HT(7) receptor probing in vitro even if the FP3 series seemed to be more specific for this receptor. These results encourage us to pursue in vivo studies. PMID- 21620534 TI - Design and synthesis of spiro derivatives of parthenin as novel anti-cancer agents. AB - Several novel spiro derivatives of parthenin (1) have been synthesized by the dipolar cycloaddition using various dipoles viz; benzonitrile oxides, nitrones and azides with exocyclic double bond of C ring (alpha-methylene-gamma butyrolactone). Majority of the compounds exhibited improved anti-cancer activity compared to the parthenin, when screened for their in vitro cytotoxicity against three human cancer cell lines viz., SW-620, DU-145 and PC-3. In vivo screening of select analog revealed improved anti-cancer activity with low mammalian toxicity as compared to parthenin. The results of the cytotoxicity pattern of these derivatives reveals the SAR of these sesquiterpinoid lactones and possible role of alpha,beta-unsaturated ketone of parthenin in inhibiting NF-kB. A mechanistic correlation of anti-cancer activity along with in vivo and western blotting experiments has been described. PMID- 21620535 TI - Synthesis, characterization and antimicrobial studies of some new pyrazole incorporated imidazole derivatives. AB - In the present study two series of novel imidazole derivatives containing substituted pyrazole moiety (3a-d and 5a-j) were synthesized. The first series were synthesized by the reaction of 3-aryl-1H-pyrazole-4-carbaldehyde thiosemicarbazones (2a-d) with DMAD and the second series by the reaction of 3 aryl-1H-pyrazole-4-carbaldehydes (1a-e) with 1,2-diketones (4a,b) in the presence of ammonium acetate. Structures of newly synthesized compounds were characterized by spectral studies. New compounds were screened for antifungal and antibacterial activities. Among the synthesized compounds, compound 3c was found to be potent antimicrobial agent. The acute oral toxicity study for the compound 3c was carried out and the experimental studies revealed that compound 3c is safe up to 3000 mg/kg and no death of animals were recorded. PMID- 21620536 TI - Recent progress in biological activities of synthesized phenothiazines. AB - This review summarizes recent medicinal chemistry investigations in vitro and in vivo in search for new phenothiazines of promising biological activities. New phenothiazine derivatives (over 50 main structures) contain dialkylaminoalkyl, cycloaminoalkyl and aminoalkyl substituents and their acyl and sulfonyl derivatives, and other substituents with varied the monocyclic (pyrazole, thiazole, oxadiazole, thiadiazole, tetrazole) and bicyclic (quinolizine, pyrazolopyrimidine, thiazolopyridine, azabicyclononane and spiro[chromanpyrimidine] heterocycles linked directly or via the alkyl chain with the thiazine nitrogen atom or with the benzene ring. The modifications of the tricyclic ring system with the bicyclic homoaromatic ring (naphthalene) and monocyclic and bicyclic azine rings (pyridine, pyrimidine, pyrazine and quinoline) led to compounds of significant biological activities. Recently obtained phenothiazines exhibit promising antibacterial, antifungal, anticancer, antiviral, anti-inflammatory, antimalarial, antifilarial, trypanocidal, anticonvulsant, analgesic, immunosuppressive and multidrug resistance reversal properties. These activities were the results of the actions of phenothiazines on biological systems via the interaction of the pharmacophoric substituent (in some cases of strict length), via the interaction of the multicyclic ring system (pi pi interaction, intercalation in DNA) and via the lipophilic character allowing the penetration through the biological membranes. The activities were examined by using various biological systems such as cell lines, bacteria, viruses, parasites, laboratory mice, rats and rabbits, and monolayer and bilayer membranes. Some mechanisms of the actions are discussed. This review shows current tendency in the phenothiazine synthesis (without synthetic routes) and reveals the phenothiazine core to be very potent pharmacophoric moiety which can be a rich source of new compounds having desirable biological activities. PMID- 21620537 TI - [Mediastinal fibrosis and mirtazapin]. AB - We report a 61-year-old woman who developed a superior vena cava syndrome due to mediastinal fibrosis. This mediastinal fibrosis resolved after the discontinuation of a chronic treatment by mirtazapin that the patient was receiving for the previous six years, suggesting a role of this treatment in the genesis of the mediastinal fibrosis. PMID- 21620538 TI - The Royal Perth Hospital method for the design and manufacture of titanium cranioplasty plates. PMID- 21620539 TI - RETRACTED: Student attrition in the Ontario Midwifery Education Programme. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 21620540 TI - Quantifying air pollution attenuation within urban parks: an experimental approach in Shanghai, China. AB - Parks with various types of vegetations played an important role in ameliorating air quality in urban areas. However, the attenuation effect of urban vegetation on levels of air pollution was rarely been experimentally estimated. This study, using seasonal monitoring data of total suspended particles (TSP), sulfur dioxide (SO(2)) and nitrogen dioxide (NO(2)) from six parks in Pudong District, Shanghai, China, demonstrated vegetations in parks can remove large amount of airborne pollutants. In addition, crown volume coverage (CVC) was introduced to characterize vegetation conditions in parks and a mixed-effects model indicated that CVC and the pollution diffusion distance were key predictors influencing pollutants removal rate. Therefore, it could be estimated by regression analysis that in summer, urban vegetations in Pudong District could contribute to 9.1% of TSP removal, 5.3% of SO(2) and 2.6% of NO(2). The results could be considered for a better park planning and improving air quality. PMID- 21620541 TI - Interspecies and spatial trends in polycyclic aromatic hydrocarbons (PAHs) in Atlantic and Mediterranean pelagic seabirds. AB - PAHs were analyzed in the liver of 5 species of pelagic seabirds (Procellariiformes) from the northeast Atlantic and the Mediterranean. The main objective was to assess the trophic and geographic trends of PAHs in seabirds to evaluate their suitability as bioindicators of chronic marine pollution by these compounds. Although higher levels of PAHs have been described in the Mediterranean compared to other oceanic regions, we did not find significant spatial patterns and observed only minor effects of the geographic origin on seabird PAHs. However, we found significant higher PAH levels in petrel compared to shearwater species, which could be related to differences in their exploitation of mesopelagic and epipelagic resources, respectively, and the vertical dynamic of PAHs in the water column. Overall, although this study enhances the need of multi-species approaches to show a more comprehensive evaluation of marine pollution, seabirds emerged as poor indicators of pelagic chronic PAH levels. PMID- 21620542 TI - Indoor metallic pollution related to mining activity in the Bolivian Altiplano. AB - The environmental pollution associated with mining and metallurgical activities reaches its greatest extent in several Andean cities and villages. Many locations in this area have accumulated through centuries a large amount of mining wastes, often disregarding the magnitude of this situation. However, in these naturally mineralized regions, there is little information available stating the exact role of mining and metallurgical industries in urban pollution. In this study, we demonstrated that the various metallic elements present in indoor dust (As, Cd, Cu, Pb, Sb, Sn, Zn) had a common origin and this contamination was increased by the proximity to the mines. Lead dust concentration was found at concerning levels for public health. In addition, wrong behaviors such as carrying mining workwear home contributed to this indoor dust pollution. Consequently, the constant exposure of the population could represent a potential health hazard for vulnerable groups, especially children. PMID- 21620543 TI - 1H NMR-based metabolomics of time-dependent responses of Eisenia fetida to sub lethal phenanthrene exposure. AB - (1)H NMR-based metabolomics was used to examine the response of the earthworm Eisenia fetida after exposure to sub-lethal concentrations of phenanthrene over time. Earthworms were exposed to 0.025 mg/cm(2) of phenanthrene (1/64th of the LC(50)) via contact tests over four days. Earthworm tissues were extracted using a mixture of chloroform, methanol and water, resulting in polar and non-polar fractions that were analyzed by (1)H NMR after one, two, three and four days. NMR based metabolomic analyses revealed heightened E. fetida responses with longer phenanthrene exposure times. Amino acids alanine and glutamate, the sugar maltose, the lipids cholesterol and phosphatidylcholine emerged as potential indicators of phenanthrene exposure. The conversion of succinate to fumarate in the Krebs cycle was also interrupted by phenanthrene. Therefore, this study shows that NMR-based metabolomics is a powerful tool for elucidating time-dependent relationships in addition to the mode of toxicity of phenanthrene in earthworm exposure studies. PMID- 21620544 TI - Nitrogen concentrations in mosses indicate the spatial distribution of atmospheric nitrogen deposition in Europe. AB - In 2005/6, nearly 3000 moss samples from (semi-)natural location across 16 European countries were collected for nitrogen analysis. The lowest total nitrogen concentrations in mosses (<0.8%) were observed in northern Finland and northern UK. The highest concentrations (>= 1.6%) were found in parts of Belgium, France, Germany, Slovakia, Slovenia and Bulgaria. The asymptotic relationship between the nitrogen concentrations in mosses and EMEP modelled nitrogen deposition (averaged per 50 km * 50 km grid) across Europe showed less scatter when there were at least five moss sampling sites per grid. Factors potentially contributing to the scatter are discussed. In Switzerland, a strong (r(2) = 0.91) linear relationship was found between the total nitrogen concentration in mosses and measured site-specific bulk nitrogen deposition rates. The total nitrogen concentrations in mosses complement deposition measurements, helping to identify areas in Europe at risk from high nitrogen deposition at a high spatial resolution. PMID- 21620545 TI - Using advanced surface complexation models for modelling soil chemistry under forests: Solling forest, Germany. AB - Various dynamic soil chemistry models have been developed to gain insight into impacts of atmospheric deposition of sulphur, nitrogen and other elements on soil and soil solution chemistry. Sorption parameters for anions and cations are generally calibrated for each site, which hampers extrapolation in space and time. On the other hand, recently developed surface complexation models (SCMs) have been successful in predicting ion sorption for static systems using generic parameter sets. This study reports the inclusion of an assemblage of these SCMs in the dynamic soil chemistry model SMARTml and applies this model to a spruce forest site in Solling Germany. Parameters for SCMs were taken from generic datasets and not calibrated. Nevertheless, modelling results for major elements matched observations well. Further, trace metals were included in the model, also using the existing framework of SCMs. The model predicted sorption for most trace elements well. PMID- 21620546 TI - Association between metabolic syndrome and the presence of kidney stones in a screened population. AB - BACKGROUND: Components of metabolic syndrome have been associated with kidney stone disease, but little evidence is available to support a relationship between metabolic syndrome and kidney stone development in healthy large screened populations. STUDY DESIGN: Cross-sectional analysis. SETTING & PARTICIPANTS: Data were obtained from 34,895 individuals who underwent general health screening tests between January 2006 and December 2006 at the Asan Medical Center. PREDICTOR: Metabolic syndrome was defined according to criteria established by the National Cholesterol Education Program Adult Treatment Panel III, American Heart Association, and National Heart, Lung, and Blood Institute. OUTCOMES & MEASUREMENTS: The presence of kidney stones was evaluated using computed tomography or ultrasonography. RESULTS: Of all those screened, 839 (2.4%) had radiologic evidence of kidney stones and metabolic syndrome was diagnosed in 4,779 (13.7%). The multivariable-adjusted OR for kidney stones increased with an increasing quintile of waist circumference and systolic/diastolic blood pressure (P < 0.001). Age, sex, hypertension, and metabolic syndrome status were independent risk factors for kidney stones. The presence of metabolic syndrome had an OR of 1.25 (95% CI, 1.03-1.50) for kidney stone prevalence. In participants with hypertension, the OR for the presence of kidney stones was 1.47 (95% CI, 1.25-1.71) compared with that for participants without hypertension after adjustment for other variables. LIMITATIONS: Cross-sectional design, absence of stone composition. CONCLUSION: Metabolic syndrome is associated with a significantly increased risk of kidney stone development. Our findings suggest the need for interventional studies to test the effects of preventing and treating metabolic syndrome on the risk of kidney stone development. PMID- 21620547 TI - Development and validation of a self-assessment tool for albuminuria: results from the Reasons for Geographic and Racial Differences in Stroke (REGARDS) Study. AB - BACKGROUND: The prevalence of albuminuria in the general population is high, but awareness of it is low. Therefore, we sought to develop and validate a self assessment tool that allows individuals to estimate their probability of having albuminuria. STUDY DESIGN: Cross-sectional study. SETTING & PARTICIPANTS: The population-based Reasons for Geographic and Racial Differences in Stroke (REGARDS) Study for model development and the National Health and Nutrition Examination Survey (NHANES) 1999-2004 for model validation. US adults 45 years or older in the REGARDS Study (n = 19,697) and NHANES 1999-2004 (n = 7,168). PREDICTOR: Candidate items for the self-assessment tool were collected using a combination of interviewer- and self-administered questionnaires. OUTCOME: Albuminuria was defined as a urinary albumin to urinary creatinine ratio >=30 mg/g in spot samples. RESULTS: 8 items were included in the self-assessment tool (age, race, sex, current smoking, self-rated health, and self-reported history of diabetes, hypertension, and stroke). These items provided a C statistic of 0.709 (95% CI, 0.699-0.720) and good model fit (Hosmer-Lemeshow chi(2)P = 0.49). In the external validation data set, the C statistic for discriminating individuals with and without albuminuria using the self-assessment tool was 0.714. Using a threshold of >=10% probability of albuminuria from the self-assessment tool, 36% of US adults 45 years or older in NHANES 1999-2004 would test positive and be recommended for screening. Sensitivity, specificity, and positive and negative predictive values for albuminuria associated with a probability >=10% were 66%, 68%, 23%, and 93%, respectively. LIMITATIONS: Repeated urine samples were not available to assess the persistency of albuminuria. CONCLUSIONS: 8 self-report items provide good discrimination for the probability of having albuminuria. This tool may encourage individuals with a high probability to request albuminuria screening. PMID- 21620548 TI - The role of the human extrastriate visual cortex in mirror symmetry discrimination: a TMS-adaptation study. AB - The human visual system is able to efficiently extract symmetry information from the visual environment. Prior neuroimaging evidence has revealed symmetry preferring neuronal representations in the dorsolateral extrastriate visual cortex; the objective of the present study was to investigate the necessity of these representations in symmetry discrimination. This was accomplished by the use of state-dependent transcranial magnetic stimulation, which combines the fine resolution of adaptation paradigms with the assessment of causality. Subjects were presented with adapters and targets consisting of dot configurations that could be symmetric along either the vertical or horizontal axis (or they could be non-symmetric), and they were asked to perform a symmetry discrimination task on the targets while fixating the center of the screen. TMS was applied during the delay between the adapter and the test stimulus over one of four different sites: Left or Right V1/V2, or left or right dorsolateral extrastriate cortex (DLO). TMS over both Left and Right DLO reduced the adaptation effect in detecting vertical and horizontal symmetry, although the Left DLO effect on horizontal symmetry and the Right DLO effect on both vertical and horizontal symmetry were present only when considering subjects who showed a behavioral adaptation effect in the baseline No-TMS condition. Application of TMS over the Left or Right V1/V2 did not modulate the adaptation effect. Overall, these data suggest that both the Left and Right DLO contain neuronal representations tuned to mirror symmetry which play a causal role in symmetry discrimination. PMID- 21620549 TI - [Dynamic nature of coronary plaques and clinical outcomes in diabetic patients: is change in itself bad?]. PMID- 21620550 TI - A comparison of the area of chest compression by the superimposed-thumb and the alongside-thumb techniques for infant cardiopulmonary resuscitation. AB - OBJECTIVES: We investigated whether the superimposed-thumb technique could reduce the chest compression area in infant cardiopulmonary resuscitation (CPR). METHODS: Charts and multidirectional computed tomography images of infants presented to four hospitals from January 2007 to September 2010 were reviewed retrospectively. We measured at the point of maximal anterior-posterior heart diameter the width of the sternum meter (S(ap)), vertical heart length from S(ap), length and width of the superimposed-thumb technique and length and width of the alongside-thumb technique. We studied the structures located underneath thumbs superimposed and thumbs alongside at S(ap) and S(nipple) (the sternum of the inter-nipple line). RESULTS: In the 84 infants enrolled, the width of the sternum at S(ap), and the vertical heart length from S(ap) were 0.85 +/- 0.31 and 1.71 +/- 0.47 cm, respectively. The length and width of the superimposed-thumb technique were 1.65 +/- 0.13 and 2.73 +/- 0.22 cm, respectively. The length and width of the alongside-thumb technique were 3.00 +/- 0.48 and 3.77 +/- 0.24 cm, respectively. The liver was situated underneath thumbs superimposed at S(ap) in 59.5% infants. The livers and lungs of 73.8% and 64.3% infants, respectively, were underneath thumbs alongside at S(nipple). CONCLUSION: In this study, we confirmed that the superimposed-thumb technique may reduce chest compression area in infant CPR. The lungs or livers were located more often underneath thumbs alongside at S(nipple) than underneath thumbs superimposed at S(ap). However, further studies are needed to validate the efficiency and safety of this technique. PMID- 21620551 TI - Beneficial effects of erythropoietin on airway histology in a murine model of chronic asthma. AB - BACKGROUND: Erythropoietin (EPO) is originally defined as a haematopoietic growth factor, but also has anti-inflammatory effects through cytokine modulation. This anti-inflammatory and cytokine modulating effect has not been investigated for the treatment of asthma. We aimed to determine the beneficial effects of erythropoietin on lung histology of murine model of chronic asthma. METHODS: Thirty-five BALB/c mice were divided into five groups: I; II; III; IV; and control group. All groups except control group were sensitised and challenged with ovalbumin. Mice with experimentally induced asthma in Group I received saline; Group II EPO 500IU/kg; Group III EPO 1000IU/kg; and Group IV dexamethasone 1mg/kg intraperitoneally once a day in the last five days of the challenge period. Animals were sacrificed 24h after the last administration of study drugs. Histological findings of airways were evaluated by light and electron microscopic examination. RESULTS: All histological parameters of asthma in the group treated with a high dose of EPO (Group III) were significantly ameliorated when compared with the group treated with saline (Group I). In comparison to the group treated with low dose of EPO (Group II) and the group treated with saline (Group I), basement membrane thicknesses and number of mast cells were significantly lower in the group treated with low dose of EPO (Group II). All histological parameters were similar between the group treated with high dose of EPO (Group III) and the group treated with dexamethasone (Group IV) except higher number of mast cells in the group treated with high dose of EPO (Group III). Additionally, the results of all histological parameters in the group treated with high dose of EPO (Group III) were significantly better when compared with the group treated with low dose of EPO (Group II). CONCLUSIONS: We found that EPO ameliorated histological changes of chronic murine model of asthma. Further studies are needed to evaluate the efficacy of EPO in the treatment of asthma. PMID- 21620552 TI - Exfoliative erythrodermia induced by pantoprazole. PMID- 21620553 TI - Comparison of re-operation rates and complication rates after total laparoscopic hysterectomy (TLH) and laparoscopy-assisted supracervical hysterectomy (LASH). AB - OBJECTIVE: To compare re-operation rates and complication rates after total laparoscopic hysterectomy (TLH) and laparoscopy-assisted supracervical hysterectomy (LASH). STUDY DESIGN: Retrospective analysis of 867 women who underwent laparoscopic hysterectomy between January 2002 and December 2009 for benign gynaecological diseases. Total laparoscopic hysterectomy was performed in 567 women (TLH group) and laparoscopy-assisted supracervical hysterectomy was performed in 300 women (LASH group). RESULTS: The women in the LASH group were significantly younger (45.6 years) than those in the TLH group (47.9 years) and the uteri removed with LASH were significantly heavier (326.4 g) than those removed with TLH (242.7 g). The rate of salpingo-oophorectomy was significantly lower in the LASH group. The overall re-operation rates were equivalent in the two groups. Two method-specific reasons for re-operations were identified. A method-specific procedure after LASH was extirpation of the cervical stump, which was performed in 2.7% of the women. Vaginal cuff dehiscence was a method-specific problem leading to secondary operation after TLH and was observed in 0.7% of the patients. No differences between the intraoperative and postoperative complication rates were observed, although there was a trend toward lower complication rates after LASH. CONCLUSIONS: There seem to be equivalent overall re-operation rates and complication rates after both hysterectomy procedures, making the two laparoscopic approaches for hysterectomy equivalent. PMID- 21620554 TI - An acquired sensation of wide/smooth vagina: a new classification. AB - OBJECTIVE: To establish a new clinical classification for a wide/smooth vagina. STUDY DESIGN: A prospective multiple time series clinical study without intervention. Vaginal rugation, and vaginal/perineal site-specific defects associated with wide/smooth vagina were used as basic clinical parameters to establish a new classification. One hundred symptomatic women associated with acquired sensations of wide/smooth vagina were enrolled. Main outcome measures were to establish the absence or presence of vaginal columnar rugae and/or site specific vaginal/perineal/vaginal orifice defects. RESULTS: The absence of columnar rugation and the absence of vaginal/perineal site-specific defects were identified in 16 subjects; the absence of columnar rugae and the presence of vaginal/perineal site-specific defects were present in 76 women. The presence of columnar rugae and the presence of vaginal/perineal site-specific defects were demonstrated in 5 subjects; and the presence of columnar rugae and absence of vaginal/perineal site-specific defects were recognized in 3 subjects. CONCLUSIONS: A wide vagina classification was developed as a "Category A, Category B, Category C or Category D". PMID- 21620555 TI - Hydatid of Morgagni: a possible underestimated cause of unexplained infertility. AB - OBJECTIVE: To evaluate the possible role of hydatid of Morgagni in patients with unexplained infertility. STUDY DESIGN: This was a non-randomized controlled trial conducted at the Department of Obstetrics and Gynecology of Sohag University, Egypt. Two hundred and thirteen patients with unexplained infertility and hydatid of Morgagni diagnosed at laparoscopy were included. The laterality (bilateral vs unilateral), location (fimbrial vs juxta-fimbrial), number (single vs multiple) and diameter of the hydatids of Morgagni were recorded. Patients were allocated to a study group (n=127) who underwent laparoscopic excision of hydatid of Morgagni and a control group (n=86) who underwent no intervention. Patients were followed for six months without any infertility or hormonal treatment to detect spontaneous pregnancy. Patients missed during the follow-up or who received infertility treatment were excluded. Statistical analysis was done using Chi square test and Student's t-test. To find the most important character of hydatid of Morgagni which impedes pregnancy, logistic regression analysis of the dependent variable (no pregnancy) and independent variables (different characters of hydatid of Morgagni) was carried out in the control group. RESULTS: Hydatid of Morgagni was detected in 52.1% of patients with unexplained infertility compared to 25.6% of those with explained infertility (p<0.001). The pregnancy rate was higher in the study group than the control group (58.7% vs 20.6%, p<0.001). The pregnancy rate was significantly higher in the study group than the control group if the hydatid cyst was bilateral (85.7% vs 5.3%, p<0.001), fimbrial (85.6% and 9.1%, p<0.001), single (57.6% and 30.3%, p<0.001) or 1-2 cm in diameter (58.1% and 25.5%, p<0.001). Logistic analysis showed that the bilaterality and fimbrial location of the hydatid of Morgagni were the most significant characteristics impeding pregnancy (odds ratio=7.27 and 3.67 respectively). CONCLUSIONS: Hydatid of Morgagni is a possible underestimated cause of unexplained infertility. Laparoscopic removal of hydatid of Morgagni in patients with unexplained infertility was followed with a high spontaneous pregnancy rate. This is particularly obvious with bilateral and fimbrial hydatid of Morgagni. PMID- 21620556 TI - Pregnancy following radical vulvectomy for carcinoma of the vulva: a case report and literature review. PMID- 21620557 TI - Methadone and the causes of maternal and perinatal complications: a response to Pinto et al. "Substance abuse during pregnancy: effect on pregnancy outcomes" [Eur. J. Obstet. Gynecol. Reprod. Biol. 150 (2010) 137-141]. PMID- 21620558 TI - Atosiban versus betamimetics in the treatment of preterm labour in Italy: clinical and economic importance of side-effects. AB - The aim of this study was to determine the cost effectiveness of atosiban compared to betamimetics in the treatment of preterm labour within the Italian setting. A systematic literature review identified randomised controlled trials (RCTs) comparing atosiban with betamimetics. Meta-analysis of nine RCTs determined that atosiban and betamimetics had similar efficacy in delaying preterm birth by at least 48 h (p=0.910). Use of atosiban was associated with significantly fewer adverse events (p<0.008). Results demonstrate that atosiban is cost-saving versus ritodrine or isoxuprine. Atosiban cost savings are ?657 per patient from the National Health Service payer's perspective; ?299 at 18 h of tocolysis to ?189 at 48 h from the hospital's perspective. The respective values versus isoxuprine were ?303 and ?199. From the combined perspective, using atosiban versus ritodrine saved from ?425 to ?316; and versus isoxuprine from ?429 to ?326. Owing to its superior safety profile, atosiban is cost-saving versus betamimetics in the treatment of preterm labour in Italy from the payer's, hospital's and combined perspectives. With the approximate 40,000 annual preterm births in Italy the annual savings could be in excess of ?13 million for the payer or ?3.8-6.2 million for the hospitals. PMID- 21620559 TI - Laparoendoscopic single-site surgery (LESS) for adnexal tumors: one surgeon's initial experience over a one-year period. AB - OBJECTIVE: In this article, we present our initial experience with laparoendoscopic single-site surgery (LESS) for adnexal tumors and report on the safety and feasibility of this technique. STUDY DESIGN: We retrospectively reviewed the medical records of the first 94 consecutive patients who underwent LESS for adnexal tumors at Cheil General Hospital and Women's Healthcare Center, Seoul, Korea between March 2009 and July 2010. Our retrospective chart review was approved by our local Institutional Review Board (IRB). We used a homemade wound retractor and surgical glove as the single-port device. All LESS procedures were performed by a single surgeon. RESULTS: Of 94 patients, 90 (95.7%) successfully underwent LESS for the treatment of an adnexal tumor. The mean age of the patients was 38.51 +/- 11.65 years, and the mean body mass index was 21.22 +/- 2.79 kg/m(2). The mean operative time was 50.33 +/- 13.26 min, and the mean diameter of the adnexal tumors was 6.26 +/- 3.19 cm. The procedures included enucleation of an ovarian cyst (n=56), salpingo-oophorectomy (n=32), and salpingectomy (n=4). Two cases with suspicion for malignancy underwent conversion to laparotomy (2.1%), and frozen sections revealed serous ovarian adenocarcinoma (n=1) and chronic inflammation (n=1). Except for those two cases, the pathologic diagnoses were confirmed as mature cystic teratoma (n=30), endometriosis (n=24), mucinous cystadenoma (n=14), serous cystadenoma (n=14), borderline ovarian tumor (n=4), hydrosalpinx (n=4), serous adenocarcinoma of the salpinx (n=1), and ectopic pregnancy (n=1). Two cases required an additional trocar for adhesiolysis of severe pelvic adhesion. No major intraoperative or postoperative complications occurred. CONCLUSIONS: Our results suggest that LESS is a safe and feasible alternative to conventional laparoscopic surgery for the treatment of adnexal tumors. PMID- 21620561 TI - Stepwise approach for nerve sparing without countertraction during robot-assisted radical prostatectomy: technique and outcomes. AB - BACKGROUND: Although subtle technical variation affects potency preservation during robot-assisted laparoscopic radical prostatectomy (RARP), most prostatectomy studies focus on achieving the optimal anatomic nerve-sparing dissection plane. However, the impact of active assistant/surgeon neurovascular bundle (NVB) countertraction on sexual function outcomes has not been studied or quantified. OBJECTIVE: To illustrate technique and compare sexual function outcomes for nerve sparing without (NS-0C) versus with (NS-C) assistant and/or surgeon NVB countertraction. DESIGN, SETTING, AND PARTICIPANTS: This is a retrospective study of 342 NS-0C versus 268 NS-C RARP procedures performed between August 2008 and February 2011. SURGICAL PROCEDURE: RARP. MEASUREMENTS: We used the Expanded Prostate Cancer Index Composite (EPIC) sexual function and potency scores, estimated blood loss (EBL), operative time, and positive surgical margin (PSM). RESULTS AND LIMITATIONS: In unadjusted analysis, men undergoing NS 0C versus NS-C were older, had worse baseline sexual function, higher biopsy and pathologic Gleason grade, and higher preoperative prostate-specific antigen (PSA) levels (all p <= 0.023). However, NS-0C versus NS-C was associated with higher 5 mo sexual function scores (20 vs 10; p < 0.001), and this difference was accentuated for bilateral intrafascial nerve sparing in preoperatively potent men (35.8 vs 16.6; p < 0.001). Similarly, 5-mo potency for preoperatively potent men was better with bilateral intrafascial NS-0C versus NS-C (45.0% vs 28.4%; p = 0.039). However, no difference in sexual function or potency was observed at 12 mo. In adjusted analyses, NS-0C versus NS-C was associated with improved 5-mo sexual function (parameter estimate: 10.90; standard error: 2.16; p < 0.001) and potency (odds ratio: 1.69; 95% confidence interval, 1.01-2.83; p = 0.046). NS-0C versus NS-WC was associated with shorter operative times (p = 0.001) and higher EBL (p = 0.001); however, there were no significant differences in PSM. Limitations include the retrospective, single-surgeon study design and smaller numbers for 12-mo comparison. CONCLUSIONS: Reliance on countertraction to facilitate dissecting NVB away from the prostate leads to neuropraxia and delayed recovery of sexual function and potency. Subtle technical modification to dissect the prostate away from the NVB without countertraction enables earlier return of sexual function and potency. PMID- 21620560 TI - Androgens and estrogens in benign prostatic hyperplasia: past, present and future. AB - Benign prostatic hyperplasia (BPH) and associated lower urinary tract symptoms (LUTS) are common clinical problems in urology. While the precise molecular etiology remains unclear, sex steroids have been implicated in the development and maintenance of BPH. Sufficient data exists linking androgens and androgen receptor pathways to BPH and use of androgen reducing compounds, such as 5alpha reductase inhibitors which block the conversion of testosterone into dihydrotestosterone, are a component of the standard of care for men with LUTS attributed to an enlarged prostate. However, BPH is a multifactorial disease and not all men respond well to currently available treatments, suggesting factors other than androgens are involved. Testosterone, the primary circulating androgen in men, can also be metabolized via CYP19/aromatase into the potent estrogen, estradiol-17beta. The prostate is an estrogen target tissue and estrogens directly and indirectly affect growth and differentiation of prostate. The precise role of endogenous and exogenous estrogens in directly affecting prostate growth and differentiation in the context of BPH is an understudied area. Estrogens and selective estrogen receptor modulators (SERMs) have been shown to promote or inhibit prostate proliferation signifying potential roles in BPH. Recent research has demonstrated that estrogen receptor signaling pathways may be important in the development and maintenance of BPH and LUTS; however, new models are needed to genetically dissect estrogen regulated molecular mechanisms involved in BPH. More work is needed to identify estrogens and associated signaling pathways in BPH in order to target BPH with dietary and therapeutic SERMs. PMID- 21620562 TI - The role of laparoscopic and robotic cystectomy in the management of muscle invasive bladder cancer with special emphasis on cancer control and complications. AB - CONTEXT: Minimally invasive radical cystectomy (MIRC) techniques for the treatment of muscle-invasive bladder cancer (BCa) are being increasingly applied. MIRC offers the potential benefits of a minimally invasive approach in terms of reduced blood loss and analgesic requirements whilst striving to provide similar oncologic efficacy to open radical cystectomy (ORC). Whether quicker recovery, shorter hospital stay, and a reduction in complications are routinely achieved with MIRC remains to be proved in prospective comparisons. OBJECTIVE: To explore both laparoscopic radical cystectomy (LRC) and robot-assisted radical cystectomy (RRC), focusing specifically on the oncologic parameters and comorbidity of the procedures. Reported complications from major centres are identified and categorised via the Clavien system. Positive margins rates, local recurrence, and both cancer-specific survival (CSS) and overall survival rates are assessed. EVIDENCE ACQUISITION: A comprehensive electronic literature search was conducted in November 2010 using the Medline database to identify publications relating to laparoscopic, robotic, or minimally invasive radical cystectomy. EVIDENCE SYNTHESIS: There are encouraging short- to medium-term results for both LRC and RRC in terms of postoperative morbidity and oncologic outcomes. It seems possible in experienced hands to perform a satisfactory minimally invasive lymphadenectomy regarding lymph node counts and levels of dissection. Positive soft-tissue margins are similar to large open series for T2/T3 disease but inferior for bulky T4 disease. Local recurrence rates and CSS rates seem equivalent to ORC at up to 3 yr of follow-up; however, mature outcome data still need to be presented before definitive comparisons can be made. CONCLUSIONS: Robotic and laparoscopic cystectomy has a growing role in the management of muscle-invasive BCa. Long-term oncologic results are awaited, and there are concerns over the ability of MIRC to treat bulky and locally advanced disease, making careful patient selection vital. Forthcoming randomised trials in this area will more fully address these issues. PMID- 21620563 TI - Slow-progressive ataxia with a methionine-to-arginine point mutation in codon 232 in the prion protein gene (PRNP). PMID- 21620564 TI - Simultaneous absorption of NOx and SO2 from flue gas with pyrolusite slurry combined with gas-phase oxidation of NO using ozone. AB - NO was oxidized into NO(2) first by injecting ozone into flue gas stream, and then NO(2) was absorbed from flue gas simultaneously with SO(2) by pyrolusite slurry. Reaction mechanism and products during the absorption process were discussed in the followings. Effects of concentrations of injected ozone, inlet NO, pyrolusite and reaction temperature on NO(x)/SO(2) removal efficiency and Mn extraction rate were also investigated. The results showed that ozone could oxidize NO to NO(2) with selectivity and high efficiency, furthermore, MnO(2) in pyrolusite slurry could oxidize SO(2) and NO(2) into MnSO(4) and Mn(NO(3))(2) in liquid phase, respectively. Temperature and concentrations of injected ozone and inlet NO had little impact on both SO(2) removal efficiency and Mn extraction rate. Specifically, Mn extraction rate remained steady at around 85% when SO(2) removal efficiency dropped to 90%. NO(x) removal efficiency increased with the increasing of ozone concentration, inlet NO concentration and pyrolusite concentration, however, it remained stable when reaction temperature increased from 20 degrees C to 40 degrees C and decreased when the flue gas temperature exceeded 40 degrees C. NO(x) removal efficiency reached 82% when inlet NO at 750 ppm, injected ozone at 900 ppm, concentration of pyrolusite at 500 g/L and temperature at 25 degrees C. PMID- 21620565 TI - Reduction of excess sludge production in sequencing batch reactor through incorporation of chlorine dioxide oxidation. AB - In this study, chlorine dioxide (ClO(2)) instead of chlorine (Cl(2)) was proposed to minimize the formation of chlorine-based by-products and was incorporated into a sequencing batch reactor (SBR) for excess sludge reduction. The results showed that the sludge disintegrability of ClO(2) was excellent. The waste activated sludge at an initial concentration of 15 g MLSS/L was rapidly reduced by 36% using ClO(2) doses of 10mg ClO(2)/g dry sludge which was much lower than that obtained using Cl(2) based on similar sludge reduction efficiency. Maximum sludge disintegration was achieved at 10mg ClO(2)/g dry sludge for 40 min. ClO(2) oxidation can be successfully incorporated into a SBR for excess sludge reduction without significantly harming the bioreactor performance. The incorporation of ClO(2) oxidation resulted in a 58% reduction in excess sludge production, and the quality of the effluent was not significantly affected. PMID- 21620566 TI - Activation of cannabinoid receptors by the pentacyclic triterpene alpha,beta amyrin inhibits inflammatory and neuropathic persistent pain in mice. AB - In this study, we report that alpha,beta-amyrin, a plant-derived pentacyclic triterpene, reduced persistent inflammatory and neuropathic hyperalgesia in mice by a direct activation of the CB(1) and CB(2) cannabinoid receptors (CB(1)R and CB(2)R). The oral treatment with alpha,beta-amyrin (30 mg/kg) significantly reduced mechanical and thermal hyperalgesia and inflammation induced by complete Freund's adjuvant (CFA) and by partial sciatic nerve ligation (PSNL). The pretreatment with either CB(1)R or CB(2)R antagonists and the knockdown gene of the receptors significantly reverted the antinociceptive effect of alpha,beta amyrin. Of note, binding studies showed that alpha,beta-amyrin directly bound with very high affinity to CB(1)R (K(i)=0.133 nM) and with a lower affinity to CB(2)R (K(i)=1989 nM). Interestingly, alpha,beta-amyrin, ACEA (CB(1)R agonist), or JWH-133 (CB(2)R agonist), at doses that caused antinociception, failed to provoke any behavioral disturbance, as measured in the tetrad assay. In addition, alpha,beta-amyrin largely decreased interleukin-1beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha), keratinocyte-derived chemokine (KC) and interleukin 6 (IL-6) levels, and myeloperoxidase activity. Likewise, alpha,beta-amyrin prevented the activation of the transcriptional factors: nuclear factor kappaB (NF-kappaB) and cyclic adenosine monophosphate response element binding (CREB) and the expression of cyclooxygenase 2 in mice footpads and spinal cords. The present results demonstrated that alpha,beta-amyrin exhibits long-lasting antinociceptive and anti-inflammatory properties in 2 models of persistent nociception via activation of cannabinoid receptors and by inhibiting the production of cytokines and expression of NF-kappaB, CREB and cyclooxygenase 2. PMID- 21620567 TI - Contribution of afferent pathways to nerve injury-induced spontaneous pain and evoked hypersensitivity. AB - A predominant complaint in patients with neuropathic pain is spontaneous pain, often described as burning. Recent studies have demonstrated that negative reinforcement can be used to unmask spontaneous neuropathic pain, allowing for mechanistic investigations. Here, ascending pathways that might contribute to evoked and spontaneous components of an experimental neuropathic pain model were explored. Desensitization of TRPV1-positive fibers with systemic resiniferatoxin (RTX) abolished spinal nerve ligation (SNL) injury-induced thermal hypersensitivity and spontaneous pain, but had no effect on tactile hypersensitivity. Ablation of spinal NK-1 receptor-expressing neurons blocked SNL induced thermal and tactile hypersensitivity as well as spontaneous pain. After nerve injury, upregulation of neuropeptide Y (NPY) is observed almost exclusively in large-diameter fibers, and inactivation of the brainstem target of these fibers in the nucleus gracilis prevents tactile but not thermal hypersensitivity. Blockade of NPY signaling within the nucleus gracilis failed to block SNL-induced spontaneous pain or thermal hyperalgesia while fully reversing tactile hypersensitivity. Moreover, microinjection of NPY into nucleus gracilis produced robust tactile hypersensitivity, but failed to induce conditioned place aversion. These data suggest that spontaneous neuropathic pain and thermal hyperalgesia are mediated by TRPV1-positive fibers and spinal NK-1-positive ascending projections. In contrast, the large-diameter dorsal column projection can mediate nerve injury induced tactile hypersensitivity, but does not contribute to spontaneous pain. Because inhibition of tactile hypersensitivity can be achieved either by spinal manipulations or by inactivation of signaling within the nucleus gracilis, the enhanced paw withdrawal response evoked by tactile stimulation does not necessarily reflect allodynia. PMID- 21620568 TI - Perspectives on canine and feline hepatozoonosis. AB - Two species of Hepatozoon are currently known to infect dogs and cause distinct diseases. Hepatozoon canis prevalent in Africa, Asia, southern Europe, South America and recently shown to be present also in the USA causes infection mainly of hemolymphoid organs, whereas Hepatozoon americanum prevalent in the southeastern USA causes myositis and severe lameness. H. americanum is transmitted by ingestion of the Gulf Coast tick Amblyomma maculatum and also by predation on infected prey. H. canis is transmitted by Rhipicephalus sanguineus, in South America also by Amblyomma ovale, and has also been shown to be transmitted transplacentally. Hepatozoonosis of domestic cats has been described mostly from the same areas where canine infection is present and the exact identity of the species which infect cats, their pathogenicity and vectors have not been elucidated. The diagnosis of hepatozoonosis is made by observation of gamonts in blood smears, histopathology, PCR or serology. The main treatment for H. canis is with imidocarb dipropionate whereas H. americanum infection is treated with an initial combination of trimethoprim-sulfadiazine, pyrimethamine and clindamycin followed by maintenance with decoquinate. Treatment for both diseases has not been reported to facilitate complete parasite elimination and new effective drugs are needed for the management of these infections. Prevention of hepatozoonosis should be based on avoidance of oral ingestion of infected tick vectors and infected prey. PMID- 21620569 TI - Detection of human filarial parasite Brugia malayi in dogs by histochemical staining and molecular techniques. AB - Human filariasis caused by Brugia malayi is still a public health problem in many countries of Asia including India, Indonesia, Malaysia and Thailand. The World Health Organization (WHO) has targeted to eliminate filariasis by the year 2020 by Mass annual single dose Diethylcarbamazine Administration (MDA). Results of the MDA programme after the first phase was less satisfactory than expected. Malayan filariasis caused by B. malayi is endemic in the south of Thailand where domestic cat serves as the major reservoir host. There is no report about the occurrence of B. malayi in dogs. The present work was carried out to find out the incidence of microfilariasis in dogs and also to detect the presence of human filarial infection in dogs, if any. One hundred dogs above 6 months of age presented to the veterinary college Hospital, Mannuthy, Kerala, with clinical signs suggestive of microfilariasis - fever, anorexia, conjunctivitis, limb and scrotal oedema - were screened for microfilariae by wet film examination. Positive cases were subjected to Giemsa staining, histochemical staining and molecular techniques. Results of the study showed that 80% of dogs had microfilariasis; out of which 20% had sheathed microfilaria. Giemsa and histochemical staining character, PCR and sequencing confirmed it as B. malayi. High prevalence of B. malayi in dogs in this study emphasized the possible role of dogs in transmission of human filariasis. PMID- 21620570 TI - "Man-ing" up and getting drunk: the role of masculine norms, alcohol intoxication and alcohol-related problems among college men. AB - Compared to college women, college men face elevated risks for problematic drinking and negative alcohol-related consequences. These risks highlight the critical need to investigate gender issues and risk factors contributing to intoxication and related problems among men. Theoretical models suggest that conforming to masculine norms or the beliefs and expectations of what it means to be a man, may help explain patterns of problematic drinking among men. The current study advances the literature by investigating the association between masculine norms, drinking to intoxication, and alcohol-related consequences among 776 undergraduate males after taking into account the importance of fraternity status and perceived peer norms. Results indicate that fraternity status and higher perceived peer norms regarding drinking increased the risks of getting drunk and experiencing alcohol-related consequences. Specifically, the masculine norms of being a "playboy", risk-taking, and winning were risk factors of drinking to intoxication; while, being a "playboy", risk-taking, and self reliance increased the risks of alcohol-related problems. Primacy of work and heterosexual presentation were two masculine norms that were protective of drinking to intoxication. Our findings contribute to important future considerations for prevention, clinical interventions, and public-health implications in college settings. PMID- 21620571 TI - The influence of co-occurring axis I disorders on treatment utilization and outcome in homeless patients with substance use disorders. AB - The current study examined the relationship between co-occurring substance use and Axis I psychiatric disorders and treatment utilization and outcome among homeless individuals (N=365) who participated in an episode of intensive outpatient substance use treatment. Compared to patients without a co-occurring diagnosis, homeless patients with a diagnosis of major depressive disorder or PTSD participated in more substance use treatment. Diagnostic status did not predict 12-month alcohol or drug treatment outcome. Substance use treatment programs appear to successfully engage homeless individuals with major depressive disorder and PTSD in treatment. Such individuals appear to achieve similar benefits from standard substance use treatment as do homeless individuals without such disorders. PMID- 21620572 TI - Reticulin fibres anchor leukaemic blasts in the marrow of patients with acute lymphoblastic leukaemia. AB - Reticulin fibrosis has been recognized in childhood ALL at diagnosis as part of the altered stromal structure in the bone marrow (BM). Increased fibre density is correlated with a higher concentration of leukaemia cells in the BM and lower numbers of blasts in peripheral blood. We hypothesize that these fibres anchor the leukaemia cells within the BM in close proximity to BM stromal cells (BMSC). The BMSC are a rich source of growth factors and cytokines which enhance leukaemia cell growth and provide protection against chemotherapy. Mobilizing the cells by breaking the 'anchoring ropes' could lead to greater exposure to apoptotic signals. PMID- 21620573 TI - Can bisphenol A diglycidyl ether (BADGE) administration prevent steroid-induced femoral head osteonecrosis in the early stage? AB - Steroid-induced osteonecrosis of the femoral head (ONFH) is associated with increase of intraosseous pressure caused by elevating of adipogenesis and fat cell hypertrophy in the bone marrow, which subsequently decreases the blood flow in the femoral head and finally resulting in bone ischemia. The early femoral head-preserving method has mainly focused on the conventional core decompression procedure. However, it only achieves a slight decrease in intra-medullary pressure with limited clinical outcome. The crucial point in prevention is to achieve a thorough decompression of intra-medullary pressure and improvement of microcirculation of the femoral head. Bisphenol-A-diglycidyl ether (BADGE), an antagonism of PPAR-gamma(Peroxisome proliferator-activated receptor gamma), has been shown to successfully reverse bone marrow adipogenesis and fat cell hypertrophy, enhances proliferation of osteoblasts, inhibit osteoclastogenesis. Therefore, we hypothesized that BADGE administration may be an appropriate novel method for the prevention of early stage steroid-induced ONFH. PMID- 21620574 TI - Diversity and safety hazards of bacteria involved in meat fermentations. AB - Food safety is a major concern for consumers and a major issue for industry which has become aware of the importance of the starter safety assessment. In the European Union, the Food Safety Authority has introduced the Qualified Presumption of Safety (QPS) approach for safety assessment of microorganisms throughout the food chain. This assessment relies on: taxonomy, familiarity, pathogenicity and end use. Productions of toxins as well as biogenic amines by food isolates are both of major concern as they can lead to food poisoning. The other important criterion is the presence of transmissible antibiotic resistance markers. This review underlined that the main hazard of bacteria involved in food fermentations concerns antibiotic resistance and particularly the presence of transferable genetic determinants that may present a risk for public health. Selection of starter strains should consider this hazard. Following the QPS approach, a list of bacteria has been acknowledged acceptable for consumption. PMID- 21620575 TI - Effect of high pressure processing on the level of some purines and pyrimidines nucleosides and bases in dry cured and cooked ham. AB - The effect of high pressure (at 600 and 900MPa) on the levels of purines and pyrimidines was evaluated in dry-cured and cooked ham. Pressurization of dry cured ham did not modify purines and pyrimidines contents. On the contrary, treatment at 600 MPa and 900MPa caused a decrease in guanosine and an increase in adenosine respectively. PMID- 21620576 TI - Reduction of verotoxigenic Escherichia coli in production of fermented sausages. AB - After a number of foodborne outbreaks of verotoxigenic Escherichia coli involving fermented sausages, some countries have imposed regulations on sausage production. For example, the US Food Safety and Inspection Service requires a 5 log(10) reduction of E. coli in fermented products. Such regulations have led to a number of studies on the inactivation of E. coli in fermented sausages by changing processing and post-processing conditions. Several factors influence the survival of E. coli such as pre-treatment of the meat, amount of NaCl, nitrite and lactic acid, water activity, pH, choice of starter cultures and addition of antimicrobial compounds. Also process variables like fermentation temperature and storage time play important roles. Though a large variety of different production processes of sausages exist, generally the reduction of E. coli caused by production is in the range 1-2 log(10). In many cases this may not be enough to ensure microbial food safety. By optimising ingredients and process parameters it is possible to increase E. coli reduction to some extent, but in some cases still other post process treatments may be required. Such treatments may be storage at ambient temperatures, specific heat treatments, high pressure processing or irradiation. HACCP analyses have identified the quality of the raw materials, low temperature in the batter when preparing the sausages and a rapid pH drop during fermentation as critical control points in sausage production. This review summarises the literature on the reduction verotoxigenic E. coli in production of fermented sausages. PMID- 21620577 TI - Effect of ground poppy seed as a fat replacer on meat burgers. AB - Poppy seed paste was used in koefte production as an animal fat replacer and efficacy of using poppy seed was investigated. The use of ground poppy seed in formulation of meat burgers had no effect on the moisture content of uncooked meat burgers; however, it did have a significant effect (p<0.05) on the fat content of meat burgers. Ground poppy seed addition significantly affected (p<0.05) cooking yield, moisture retention, and fat retention of meat burgers. Ground poppy seed addition significantly decreased (p<0.05) saturated fatty acid contents as the amount of ground poppy seed increased in meat burgers. A strong significant decrease (p<0.05) on the cholesterol content of meat burgers with ground poppy seed addition was observed. Samples having 20% ground poppy seed had significantly better texture and juiciness score (p<0.05) than any other sample which could be explained by increased moisture retention. Replacing animal fat with ground poppy seed paste is effective and may pave the way for an innovative meat product. PMID- 21620578 TI - Histological analysis of esophageal muscular layers from 27 autopsy cases with mixed connective tissue disease (MCTD). AB - Esophageal symptoms in mixed connective tissue disease (MCTD) have been investigated radiologically. We investigated the esophageal lesions in MCTD histopathologically, and analyzed relationships between these lesions and autoantibodies extracted from the serum of MCTD patients. Esophageal tissues from 27 MCTD patients submitted to autopsy were examined. We compared histopathological features of the esophagus in different wall layers from the mucosa, submucosa, and muscular layer to the adventitia, and in the upper, middle, and lower portions of esophagus. The most striking change observed was severe atrophy and occasional loss of smooth muscle cells in the muscular layer, followed by fibrosis. These muscular changes were particularly prominent in the inner layer of the lower esophagus. Immunohistochemically, degenerated muscular tissues of the esophagus were positive for anti-IgG and anti-C3 antibodies, but not for anti-IgM antibodies. IgG fractions extracted from three MCTD patients were immunohistochemically used to examine whether some antibodies in MCTD patients showed reactivity for esophageal components. The IgG fractions isolated from MCTD patients reacted with smooth muscle from non-connective tissue disease cases, suggesting that some serum antibodies may trigger esophageal changes. These findings suggest that esophageal lesions associated with clinical dysphagia in MCTD may be related to autoantibodies. PMID- 21620579 TI - Rectal bleeding after high-dose-rate brachytherapy combined with hypofractionated external-beam radiotherapy for localized prostate cancer: the relationship between dose-volume histogram parameters and the occurrence rate. AB - PURPOSE: To determine the predictive risk factors for Grade 2 or worse rectal bleeding after high-dose-rate brachytherapy (HDR-BT) combined with hypofractionated external-beam radiotherapy (EBRT) for prostate cancer using dose volume histogram analysis. METHODS AND MATERIALS: The records of 216 patients treated with HDR-BT combined with EBRT were analyzed. The treatment protocols for HDR-BT were 5 Gy * five times in 3 days or 7 Gy * three, 10.5 Gy * two, or 9 Gy * two in 2 days. The EBRT doses ranged from 45 to 51 Gy with a fractional dose of 3 Gy. RESULTS: In 20 patients Grade 2 or worse rectal bleeding developed, and the cumulative incidence rate was 9% at 5 years. By converting the HDR-BT and EBRT radiation doses into biologic effective doses (BED), the BED(3) at rectal volumes of 5% and 10% in the patients who experienced bleeding were significantly higher than those in the remaining 196 patients. Univariate analysis showed that a higher rectal BED(3-5%) and the use of fewer needles in brachytherapy were correlated with the incidence of bleeding, but BED(3-5%) was found to be the only significant factor on multivariate analysis. CONCLUSIONS: The radiation dose delivered to small rectal lesions as 5% is important for predicting Grade 2 or worse rectal bleeding after HDR-BT combined with EBRT for prostate cancer. PMID- 21620580 TI - Feasibility and efficacy of induction docetaxel, cisplatin, and 5-fluorouracil chemotherapy combined with cisplatin concurrent chemoradiotherapy for nonmetastatic Stage IV head-and-neck squamous cell carcinomas. AB - PURPOSE: To report the experience of treating selected fit patients with locally advanced head-and-neck squamous cell carcinoma with three cycles of induction TPF (docetaxel 75 mg/m(2), cisplatin 75 mg/m(2), 5-fluorouracil 750 mg/m(2), Days 2 5) followed by concurrent three-weekly bolus cisplatin 100 mg/m(2) chemoradiotherapy. METHODS AND MATERIALS: Between March 2006 and February 2010, 66 patients with nonmetastatic Stage IV head-and-neck squamous cell carcinoma were treated in a single institution with three cycles of induction TPF, followed by radical radiotherapy with concurrent cisplatin 100 mg/m(2). RESULTS: Median age was 54 years (range, 33-69 years). Median follow-up was 21 months (range, 4 55 months). During TPF, Grade 3 toxicity occurred in 18 patients (27%), dose modifications in 10 (15%), delays in 3 (5%), and unplanned admissions in 6 (9%); a clinical tumor response was documented in 60 patients (91%). Median time from the final cycle of TPF to commencing radiotherapy was 22 days. Sixty-two patients (94%) received radical radiotherapy, and all completed treatment with no delays >=3 days. One, two, and three cycles of concurrent cisplatin were delivered to 18 patients (29%), 38 patients (61%), and 3 patients (5%), respectively. Ninety-two percent of patients received enteral feeding; median weight loss during treatment was 7%. Forty-two patients (68%) had unplanned admissions with no on-treatment deaths. Three unrelated deaths occurred after treatment. At 1 year after treatment, 21% of patients without disease progression remained gastrostomy dependent. Of 58 assessable patients, 50 (86%) achieved a complete response after treatment. One- and 2-year progression-free survival, cause-specific survival, and overall survival were 88%, 92%, and 86% and 80%, 85%, and 80%, respectively. CONCLUSION: The combination of induction TPF with concurrent cisplatin chemoradiotherapy in patients with locally advanced head and neck squamous cell carcinoma is tolerable, with encouraging efficacy. PMID- 21620581 TI - Cone-beam CT-based delineation of stereotactic lung targets: the influence of image modality and target size on interobserver variability. AB - PURPOSE: It is generally agreed that the safe implementation of stereotactic body radiotherapy requires image guidance. The aim of this work was to assess interobserver variability in the delineation of lung lesions on cone-beam CT (CBCT) images compared with CT-based contouring for adaptive stereotactic body radiotherapy. The influence of target size was also evaluated. METHODS AND MATERIALS: Eight radiation oncologists delineated gross tumor volumes in 12 patient cases (non-small cell lung cancer I-II or solitary metastasis) on planning CTs and on CBCTs. Cases were divided into two groups with tumor diameters of less than (Group A) or more than 2 cm (Group B). Comparison of mean volumes delineated by all observers and range and coefficient of variation were reported for each case and image modality. Interobserver variability was assessed by means of standard error of measurement, conformity index (CI), and its generalized observer-independent approach. The variance between single observers on CT and CBCT images was measured via interobserver reliability coefficient. RESULTS: Interobserver variability on CT images was 17% with 0.79 reliability, compared with 21% variability on CBCT and 0.76 reliability. On both image modalities, values of the intraobserver reliability coefficient (0.99 for CT and 0.97 for CBCT) indicated high reproducibility of results. In general, lower interobserver agreement was observed for small lesions (CI(genA) = 0.62 +/- 0.06 vs. CI(genB) = 0.70 +/- 0.03, p < 0.05). The analysis of single patient cases revealed that presence of spicules, diffuse infiltrations, proximity of the tumors to the vessels and thoracic wall, and respiration motion artifacts presented the main sources of the variability. CONCLUSION: Interobserver variability for Stage I-II non-small cell lung cancer and lung metastasis was slightly higher on CBCT compared with CT. Absence of significant differences in interobserver variability suggests that CBCT imaging provides an effective tool for tumor localization, and image data could be also used for target volume delineation purposes. PMID- 21620582 TI - Determinants of complications and outcome in high-risk squamous cell head-and neck cancer treated with perioperative high-dose rate brachytherapy (PHDRB). AB - PURPOSE: To determine the impact of a set of patient, tumor, and treatment factors on toxicity and outcome in patients with head-and-neck squamous cell cancer treated with surgical resection and perioperative high-dose rate brachytherapy (PHDRB) alone (single-modality [SM] group) (n = 46) or PHDRB combined with postoperative radiation or chemoradiation (combined-modality [CM] group) (n = 57). METHODS AND MATERIALS: From 2000 to 2008, 103 patients received PHDRB after complete macroscopic resection. SM patients received 32 or 40 Gy of PHDRB in 8 or 10 twice-daily treatments for R0 and R1 resections. CM patients received 16 or 24 Gy of PHDRB in 4 or 6 twice-daily treatments for R0 and R1 resections, followed by external radiation of 45 Gy in 25 fractions with or without concomitant chemotherapy. RESULTS: Grade >=4 complications according to the Radiation Therapy Oncology Group were more frequent in the SM group than in the CM group (p = 0.024). Grade >=3 and >=4 complications increased with the antecedent of prior irradiation (p = 0.032 and p = 0.006, respectively) and with TV(150) values of 13 mL or greater (p = 0.032 and p = 0.032, respectively). After a median follow-up of 34.8 and 60.8 months for SM and CM patients, respectively, patients with high-risk margins had a 9-year local control rate of 68.0% whereas patients with wider margins had a 9-year local control of 93.7% (p = 0.045). Patients with primary and recurrent tumors had 9-year actuarial locoregional control rates of 81.8% and 54.2%, respectively (p = 0.003). Patients with lymph vascular space invasion (LVSI)-positive and LVSI-negative tumors had 9-year distant control rates of 62.8% and 81.6%, respectively (p = 0.034). Disease-free survival rates decreased in recurrent cases (p = 0.006) as well as in LVSI positive patients (p = 0.035). CONCLUSIONS: The complications observed are largely attributable to the antecedent of prior irradiation but can possibly be minimized by meticulous mapping and exhaustive planning to reduce TV(150) values. Patients with high-risk margins, LVSI-positive status, and recurrent disease have a higher risk of treatment failure, and therefore risk-directed treatment strategies are required. PMID- 21620583 TI - Outcomes after whole brain reirradiation in patients with brain metastases. AB - PURPOSE: Patients with brain metastases are often treated with whole brain radiation therapy (WBRT) for purposes of palliation. The treatment of those who experience subsequent intracranial disease progression can include a second course of WBRT, although there is controversy surrounding its safety and efficacy. This study examines the outcomes in patients at Massachusetts General Hospital who underwent reirradiation. PATIENTS AND METHODS: We examined the medical records of 17 patients at Massachusetts General Hospital with brain metastases who were initially treated with WBRT between 2002 and 2008 and were subsequently retreated with a second course of WBRT. The median dose for the first course of WBRT was 35 Gy (range, 28-40 Gy), with a fraction size of 2 to 3 Gy (median, 2.5 Gy). The median dose at reirradiation was 21.6 Gy (range, 14-30 Gy), with a fraction size of 1.5 to 2 Gy (median, 1.8 Gy). RESULTS: The second course of WBRT was administered upon radiographic disease progression in all patients. Of 10 patients with complete follow-up data, 8 patients experienced complete or partial symptom resolution, and 2 did not show clinical improvement. The time to radiographic progression was 5.2 months. The median overall survival for all patients after diagnosis of metastases was 24.7 months. The median survival time after initiation of reirradiation was 5.2 months (95% CI, 1.3-8.7). In 6 patients with stable extracranial disease, the median survival time after retreatment was 19.8 months (95% CI, 2.7-infinity), compared with 2.5 months (95% CI, 0.8-5.5) for those with extracranial disease progression (p = 0.05). Acute adverse reactions occurred in 70.5% of patients but were mild to moderate in severity. CONCLUSION: In select patients and especially those with stable extracranial disease, reirradiation may be an appropriate and effective intervention to provide symptomatic relief and slow intracranial disease progression. Side effects were minimal and did not cause substantial changes in quality of life. PMID- 21620584 TI - Megavoltage cone beam computed tomography dose and the necessity of reoptimization for imaging dose-integrated intensity-modulated radiotherapy for prostate cancer. AB - PURPOSE: Megavoltage cone beam computed tomography (MV-CBCT) dose can be integrated with the patient's prescription. Here, we investigated the effects of imaging dose and the necessity for additional optimization when using intensity modulated radiotherapy (IMRT) to treat prostate cancer. METHODS AND MATERIALS: An arc beam mimicking MV-CBCT was generated using XiO (version 4.50; Elekta, Stockholm, Sweden). The monitor units (MU) for dose calculation were determined by conforming the calculated dose to the dose measured using an ionization chamber. IMRT treatment plans of 22 patients with prostate cancer were retrospectively analyzed. Arc beams of 3, 5, 8, and 15 MU were added to the IMRT plans, and the dose covering 95% of the planning target volume (PTV) was normalized to the prescribed dose with (reoptimization) or without optimization (compensation). RESULTS: PTV homogeneity and conformality changed negligibly with MV-CBCT integration. For critical organs, an imaging dose-dependent increase was observed for the mean rectal/bladder dose (D(mean)), and reoptimization effectively suppressed the D(mean) elevations. The bladder generalized equivalent uniform dose (gEUD) increased with imaging dose, and reoptimization suppressed the gEUD elevation when 5- to 15-MU CBCT were added, although rectal gEUD changed negligibly with any imaging dose. Whereas the dose elevation from the simple addition of the imaging dose uniformly increased rectal and bladder dose, the rectal D(mean) increase of compensation plans was due mainly to low-dose volumes. In contrast, bladder high-dose volumes were increased by integrating the CBCT dose, and reoptimization reduced them when 5- to 15-MU CBCT were added. CONCLUSION: Reoptimization is clearly beneficial for reducing dose to critical organs, elevated by addition of high-MU CBCT, especially for the bladder. For low MU CBCT aimed at bony structure visualization, compensation is sufficient. PMID- 21620585 TI - Effect of facility on the operative costs of distal radius fractures. AB - PURPOSE: The purpose of this study was to investigate whether ambulatory surgery centers can deliver lower-cost care and to identify sources of those cost savings. METHODS: We performed a cost identification analysis of outpatient volar plating for closed distal radius fractures at a single academic medical center. Multiple costs and time measures were taken from an internal database of 130 consecutive patients and were compared by venue of treatment, either an inpatient facility or an ambulatory, stand-alone surgery facility. The relationships between total cost and operative time and multiple variables, including fracture severity, patient age, gender, comorbidities, use of bone graft, concurrent carpal tunnel release, and surgeon experience, were examined, using multivariate analysis and regression modeling to identify other cost drivers or explanatory variables. RESULTS: The mean operative cost was considerably greater at the inpatient facility ($7,640) than at the outpatient facility ($5,220). Cost drivers of this difference were anesthesia services, post-anesthesia care unit, and operating room costs. Total surgical time, nursing time, set-up, and operative times were 33%, 109%, 105%, and 35% longer, respectively, at the inpatient facility. There was no significant difference between facilities for the additional variables, and none of those variables independently affected cost or operative time. CONCLUSIONS: The only predictor of cost and time was facility type. This study supports the use of ambulatory stand-alone surgical facilities to achieve efficient resource utilization in the operative treatment of distal radius fractures. We also identified several specific costs and time measurements that differed between facilities, which can serve as potential targets for tertiary facilities to improve utilization. TYPE OF STUDY/LEVEL OF EVIDENCE: Economic and Decisional Analysis III. PMID- 21620586 TI - Metacarpophalangeal hyperextension deformity associated with trapezial-metacarpal arthritis. PMID- 21620587 TI - [Fetoscopic laser coagulation in 100 consecutive monochorionic pregnancies with severe twin-to-twin transfusion syndrome]. AB - OBJECTIVES: To report pre- and post-surgical datas of large series of severe twin to-twin transfusion syndrome (TTTS) managed with laser ablation surgery in our centre, to evaluate the incidence of complications, perinatal outcome and to compare with other cohorts. PATIENTS AND METHODS: Observational study of 100 cases of TTTS consecutively treated with fetoscopic laser coagulation between January 2004 and April 2010 in CMCO-SIHCUS of Schiltigheim. RESULTS: There are nine stage I, 49 stage II, 38 stage III and four stage 4. Median gestation at time of laser is 20.6 weeks (14-29) whereas median gestation at delivery is 32.6 weeks (16.3-39). Overall perinatal survival rate is 68.5% (137 children over 200). Eighty-five percent have one or more surviving twins. The survival rate is the same for donors and for recipients. Preterm premature rupture of the membranes are observed in 17% of cases and the median gestational age for this complication is 30 weeks (20-34). Cerebral abnormalities are present in 7% of newborns. CONCLUSION: Our results for the management of severe TTTS are comparable to the other reported series. There are still many questions remaining concerning the optimal management of TTTS. PMID- 21620588 TI - [HPV vaccines: the current disorder and the absolute necessity of returning to basics]. PMID- 21620589 TI - [Histopathology of coeliac disease]. AB - Coeliac disease is a common disease, affecting 1% of the population. Clinical manifestations are multiple. The diagnosis requires serologic testing and a duodenal biopsy that shows the characteristic findings of intraepithelial lymphocytosis, crypt hyperplasia and villous atrophy, and a positive response to a gluten-free diet. In most patients, the histological diagnosis is easily established. Pitfalls in the pathological diagnosis include a poorly orientated biopsy specimen, either an inadequate biopsy sampling in patients with patchy villous atrophy and the other causes of villous atrophy. A non-response to the gluten-free diet needs to reassess first, the initial diagnosis, second to be sure of the gluten-free diet adherence, and third, to exclude malignant complications such as refractory celiac disease or enteropathy-associated T-cell lymphoma. PMID- 21620590 TI - [The connective tissues, from the origin of the concept to its "Maturation" to extracellular matrix. Application to ocular tissues. Contribution to the history of medical sciences]. AB - The "Tissue" concept emerged apparently in the medical literature at about the French revolution, during the second half of the 18(th) century. It was found in the texts written by the physicians of Bearn and Montpellier, the Bordeu-s and also by the famous physician, Felix Vicq d'Azyr, the last attending physician of the queen Marie-Antoinette, "Bordeu et al. (1775) et Pouliquen (2009)". It was elaborated into a coherent doctrine somewhat later by Xavier Bichat, considered as the founder of modern pathological anatomy, Bichat. With the advent of histochemistry, from the beginning of the 20(th) century, several of the principal macromolecular components of connective tissues, collagens, elastin, "acid mucopolysaccharides" (later glycosaminoglycans and proteoglycans) and finally structural glycoproteins were characterized. These constituents of connective tissues were then designated as components of the extracellular matrix (ECM), closely associated to the cellular components of these tissues by adhesive (structural) glycoproteins as fibronectin, several others and cell receptors, "recognising" ECM-components as integrins, the elastin-receptor and others. This molecular arrangement fastens cells to the ECM-components they synthesize and mediates the exchange of informations between the cells to the ECM (inside-out) and also from the ECM-components to the cells (outside-in). This macromolecular arrangement is specific for each tissue as a result of the differentiation of their cellular components. It is also the basis and condition of the fulfillment of the specific functions of differentiated tissues. This is a short description of the passage of the "tissue" concept from its vague origin towards its precise identification at the cellular and molecular level up to the recognition of its functional importance and its establishment as an autonomous science. This can be considered as a new example of the importance of metaphors for the progress of science, Keller (1995). PMID- 21620591 TI - Primary outcomes in two randomized controlled trials of treatments for cannabis use disorders. AB - BACKGROUND: While several randomized controlled trials evaluating a range of treatments for cannabis use disorders have appeared in recent years, these have been marked by inconsistency in selection of primary outcomes, making it difficult to compare outcomes across studies. METHOD: With the aim of identifying meaningful and reliable outcome domains in treatment studies of cannabis use disorders, we evaluated multiple indicators of marijuana use, marijuana problems, and psychosocial functioning from two independent randomized controlled trials of behavioral treatments for cannabis use disorders (Ns=450 and 136). RESULTS: Confirmatory factor analysis indicated that the best-fitting model of outcomes in both trials encompassed three distinct factors: frequency of marijuana use, severity of marijuana use, and psychosocial functioning. In both trials, frequency of marijuana use and longest period of abstinence during treatment were most strongly associated with outcome during follow-up. Using two categorical definitions of "clinically significant improvement," individuals who demonstrated improvement differed on most end-of-treatment and long-term outcomes from those who did not improve. CONCLUSIONS: Results may guide future randomized controlled trials of treatments for cannabis use disorders in the collection of relevant end of-treatment outcomes and encourage consistency in the reporting of outcomes across trials. PMID- 21620592 TI - Genetic analysis of the matrix and non-structural genes of equine influenza virus (H3N8) from epizootic of 2008-2009 in India. AB - India faced an epizootic of equine influenza in 2008-2009. The isolated viruses were typed as H3N8 and grouped with the clade 2 viruses of Florida sublineage on the basis of haemagglutinin (HA) gene sequence analysis. This report describes the genetic analysis and selection pressure of matrix (M) and non-structural 1 (NS1) genes of the Indian isolates. All isolates shared 98.41% and 99.54% homology with other clade 2 viruses of Asian origin for M1 and M2 amino acid (aa) sequences, respectively. There were 3 and 4 unique aa residue changes respectively in M1 and M2 proteins in all Asian isolates. Phylogenetic analysis revealed clustering of Indian and Chinese isolates in a separate group designated here as Asian clade for M gene. Indian and Chinese isolates shared homology ranging from 98.17% to 99.08% at aa level. The M and NS1 genes were under negative selection pressure with estimated magnitude of pressure (omega) 0.054, 0.581 and 0.30 for M1, M2 and NS1, respectively. PMID- 21620593 TI - Predictors of growth velocity in early infancy in a resource-poor setting. AB - OBJECTIVE: To determine the pattern and predictors of growth velocity in early infancy in a resource-poor setting. METHODS: Weight velocity between birth and first postnatal visit was determined in a cohort of preterm and full-term infants in Lagos, Nigeria using three mathematical methods reported in the literature. Maternal and infant factors predictive of weight velocity were identified by multiple linear regression analysis. RESULTS: Overall, 658 infants were enrolled with mean gestational age of 37.7+/-2.0 weeks, birthweight of 3.2+/-0.6 kg and median age of 45 (interquartile range: 42-48) days at follow-up. Offspring of older and HIV-positive mothers had significantly lower mean weight velocities while male infants and those with low birthweight and fetal growth restriction had significantly higher mean weight velocity than their peers. These patterns were consistent across the three growth models. Maternal age (p=0.004), antenatal care (p=0.007), HIV-status (p=0.008) and gender (p<0.001) were predictive of weight velocity. Higher weight velocity was strongly associated with lower birthweight (p<0.001) indicative of "catch-up" growth as well as with higher gestational age (p<0.001). CONCLUSIONS: While maternal status is predictive of early growth faltering, preterm infants warrant timely intervention to forestall/minimise the potential health and developmental consequences associated with their sub-optimal growth trajectory. PMID- 21620594 TI - Spinal nerve root beta-APP staining in infants is not a reliable indicator of trauma. AB - This preliminary communication describes seven babies with beta-amyloid precursor protein (betaAPP) positive axonal swellings in nerve roots at multiple levels of the spinal cord. All seven babies died of natural causes. Two died in utero providing evidence for nerve root injury in the absence of trauma, two died within one day of birth and the possibility of birth related injury has to be considered. Three babies were over one month of age and had no history or pathological evidence of trauma. These findings show that if axonal injury is carefully sought in every infant death, not just in babies where trauma is suspected, it will be found in a proportion of babies dying from natural diseases. While spinal nerve root axonal injury in infants may suggest trauma, it is not, in itself, diagnostic of trauma. PMID- 21620595 TI - Relative effects of posture and activity on human height estimation from surveillance footage. AB - Height estimations based on security camera footage are often requested by law enforcement authorities. While valid and reliable techniques have been established to determine vertical distances from video frames, there is a discrepancy between a person's true static height and their height as measured when assuming different postures or when in motion (e.g., walking). The aim of the research presented in this report was to accurately record the height of subjects as they performed a variety of activities typically observed in security camera footage and compare results to height recorded using a standard height measuring device. Forty-six able bodied adults participated in this study and were recorded using a 3D motion analysis system while performing eight different tasks. Height measurements captured using the 3D motion analysis system were compared to static height measurements in order to determine relative differences. It is anticipated that results presented in this report can be used by forensic image analysis experts as a basis for correcting height estimations of people captured on surveillance footage. PMID- 21620596 TI - Primer composition and memory effect of weapons--some trends from a systematic approach in casework. AB - Since 2008, our laboratory has adopted a systematic approach to the examination of gunshot residues (GSR) in casework by analysing, whenever possible, the inorganic composition present in ammunition (cartridge cases and unused ammunition). By compiling the results of these analyses in a database, it is possible to observe some trends during the period of interest: on the one hand, the prevalence of primers containing lead, barium and antimony is about 50%, and even as high as 70% when including lead-barium-antimony based primers also containing tin; on the other hand, the prevalence of non-toxic primers is for the time being very low. Still using the same approach, test firings were performed with recovered weapons and litigious ammunition whenever possible in order to estimate the influence of the well known "memory effect" of the weapons on the GSR analysis results. The first results show a quite strong memory effect for the .22 and the .32 caliber, unlike the .38 caliber. This is probably due to a high prevalence of lead-barium-antimony based primers for the latter caliber. PMID- 21620597 TI - Suture granuloma showing false-positive finding on PET/CT after head and neck cancer surgery. AB - We report herein two cases of suture granuloma showing focal intense 2-deoxy-2 [(18)F]fluoro-d-glucose (FDG)-uptake in positron emission tomography (PET). Follow-up FDG-PET after surgical intervention for head and neck cancer revealed lesions with high FDG-uptake, which were highly suspected of being a recurrent tumor. The lesions were subjected to excisional biopsy for definitive diagnosis. Histopathological examination proved them to be suture granulomas caused by non absorbable silk sutures. It should be emphasized that suture granulomas can show false-positive findings on FDG-PET, thus requiring differential diagnosis from recurrent tumors. PMID- 21620598 TI - MRI spectrum of bone changes in the diabetic foot. AB - PURPOSES: (1) To assess the prevalence of bone marrow changes in the diabetic foot and (2) to discuss the clinical significance of these changes. METHODS: 85 patients with radiographic and magnetic resonance imaging (MRI) foot examinations were selected. Inclusion criteria were clinical diagnosis of diabetes and bone changes on radiographs and MRI. The material was selected from the image storage (PACS) system. We searched for vascular (infarct and necrosis), traumatic (bruise and occult fractures), destruction and debris, dislocation, osteochondritis, osteomyelitis. Five patients had bilateral examinations. A total of 90 feet were evaluated. RESULTS: From 90 feet, 17 (18.9%) presented with vascular changes, from them, 11 feet had infarct and 6 feet had necrosis. Twenty (22.2%) feet had traumatic changes; of them, 10 (50%) had edema on MRI. Five (25%) cases had occult fracture on MRI; and 5 (25%) had visible fracture on both X-ray and MRI. Bone destruction was detected in 8 (8.9%) feet. Bony debris was visualized in three of them. Bone dislocation was visualized in 11 (12.2%) feet. There was evidence of osteochondritis in twenty-four (26.7%) feet. Osteomyelitis was diagnosed in ten (11.1%) feet. CONCLUSION: Diabetic foot is a challenge for both clinicians and radiologists due to its complexity. The bone derangements inherent to the diabetic foot can be evaluated with high accuracy with MRI. PMID- 21620599 TI - Contrast-enhanced MDCT gastrography for detection of early gastric cancer: Initial assessment of "wall-carving image", a novel volume rendering technique. AB - OBJECTIVE: We developed a new volume rendering technique, the CT gastrography wall carving image (WC) technique, which provides a clear visualization of localized enhanced tumors in the gastric wall. We evaluated the diagnostic performance of the WC as an adjunct to conventional images in detecting early gastric cancer (EGC). MATERIALS AND METHODS: Thirty-nine patients with 43 EGCs underwent contrast-enhanced MDCT gastrography for preoperative examination. Two observers independently reviewed the images under three different conditions: term 1, Axial CT; term 2, Axial CT, MPR and VE; and term 3, Axial CT, MPR, VE and WC for the detection of EGC. The accuracy of each condition as reviewed by each of the two observers was evaluated by receiver operating characteristic analysis. Interobserver agreement was calculated using weighted-kappa statistics. RESULTS: The best diagnostic performance and interobserver agreement were obtained in term 3. The AUCs of the two observers for terms 1, 2, and 3 were 0.63, 0.73, and 0.84, and 0.57, 0.73, and 0.76, respectively. The interobserver agreement improved from fair at term 1 to substantial at term 3. CONCLUSIONS: The addition of WC to conventional MDCT display improved the diagnostic accuracy and interobserver reproducibility for the detection of ECG. WC represents a suitable alternative for the visualization of localized enhanced tumors in the gastric wall. PMID- 21620600 TI - Predictive value of small ulcers in the evolution of acute type B intramural hematoma. AB - OBJECTIVE: To assess the long-term evolution and predictive factors of type B intramural hematoma (IMH). MATERIAL AND METHODS: 34 patients (33 men), mean age: 67 years (47-87) diagnosed with type B IMH by computed tomography (CT) and followed up clinically and by CT yearly. Mean follow-up was 5.9 years (2-13 years). Two evolution patterns were considered: (a) regression and (b) progression. Clinical and imaging variables were analyzed for assessing their predictor values. RESULTS: Evolution at one year was to regression in 56% and to progression in 44% of cases. There were no association among age, sex, other aortic abnormalities, presence of atherosclerotic disease or blood pressure, initial maximum aortic diameter, indexed maximum aortic diameter, IMH thickness or length, presence or absence of mediastinal hematoma and the evolution of type B IMH. Ten patients had small aortic ulcers in the acute phase. The presence of ulcers was related with progression of IMH. No differences were observed in evolution between the control at first year and the last control. CONCLUSIONS: The presence of small ulcers is a strong predictor of evolution in acute type B IMH. In addition, the regression group remains completely stable after the first year of evolution. PMID- 21620601 TI - Adrenal venous sampling using Dyna-CT--a practical guide. AB - Primary hyperaldosteronism due to aldosterone secreting adrenal adenomas is an important and potentially curable cause for hypertension. The differentiation between unilateral or bilateral adrenal adenomas is crucial, as unilateral adenomas can easily be cured by surgery whereas bilateral adenomas have to be treated conservatively. Exact diagnosis can be made when unilateral or bilateral hormone production is proven with adrenal vein sampling. We present an effective step-by-step technique how to perform an adrenal vein sampling with a special emphasis on how to reliably catheterize the right adrenal vein using Dyna CT. PMID- 21620602 TI - Development of a two-step tier-2 dissolution method for blinded overencapsulated erlotinib tablets using UV fiber optic detection. AB - Measuring dissolution of a comparator drug overencapsulated in a hard gelatin shell is necessary when determining performance of the native and blinded formulations. However, the gelatin in the shell may form cross-links upon storage at stressed conditions, resulting in slow dissolution of the encapsulated drug. The aim of this study was to develop a dissolution approach for a hard-gelatin overencapsulated formulation of a comparator drug, erlotinib, which can overcome cross linking of the capsule shell. In this case, following the USP two-tier dissolution test by simply adding an enzyme did not dissolve the cross-linked capsules because the medium used in the method for erlotinib described in the FDA Dissolution Database contains sodium dodecyl sulfate that inhibits the activity of the enzyme. Changing the method by using different surfactants was not considered acceptable because it is preferable to closely follow the compendial method for the comparator. A two-step tier-2 method was developed as a solution, without significant change to the compendial method conditions. It uses 0.1N HCl + pepsin as the initial medium to help capsule break-up. SDS is added at 15 min after the testing starts to ensure dissolution of the drug. This may be a useful general approach for dealing with cross-linking in over-encapsulated comparators. A UV fiber optic spectrophotometer was used for in situ, real-time detection of the dissolution profile during method development studies. The fast sampling rate available with this type of detection was important in elucidating the events occurring during dissolution and determining the optimal time of the SDS addition. PMID- 21620603 TI - Identification of thymol phase I metabolites in human urine by headspace sorptive extraction combined with thermal desorption and gas chromatography mass spectrometry. AB - Development of a novel highly sensitive headspace sorptive extraction (HSSE) method in combination with thermal desorption gas chromatography coupled to a mass spectrometer (TD-GC/MS) allowed the identification of thymol and several phase I metabolites in human urine. Combined with an enzymatic hydrolysis of glucuronated or sulphated phase II metabolites of thymol and of the respective phase I metabolites prior to analysis, even trace quantities of hitherto not detected thymol phase I metabolites could be identified in urine samples of test persons after oral administration of 50mg thymol. It was proven, that human metabolism leads to a hydroxylation of the aromatic ring as well as of the iso propyl side chain. Hydroxylation of the iso-propyl group results in the formation of the rather unstable p-cymene-3,8-diol and the corresponding dehydration product p-cymene-3-ol-8-ene which could be clearly detected in human urine samples. Furthermore, the aromatic hydroxylation products p-cymene-2,5-diol, its oxidation product p-cymene-2,5-dione and p-cymene-2,3-diol were also unambiguously identified by comparison with synthesized reference compounds. PMID- 21620604 TI - Effect of the traditional Chinese medicine tongxinluo on endothelial dysfunction rats studied by using urinary metabonomics based on liquid chromatography-mass spectrometry. AB - A urinary metabonomic method based on ultra-fast liquid chromatography coupled with ion trap-time of flight mass spectrometry (UFLC/MS-IT-TOF) was employed to study the preventive efficacy and the metabolic changes caused by simavastatin and the traditional Chinese medicine tongxinluo in endothelial dysfunction rats. Principal component analysis (PCA) was applied to study metabolic patterns of endothelial dysfunction rats and healthy control rats. 1-Methyladenosine, indoxyl sulfate, hippuric acid, riboflavin, coproporphyrin, and p-cresol glucuronide were identified as potential biomarkers, indicating that pathways of adenine, tryptophan, phenylalanine, riboflavin and porphyrin metabolism were disturbed in endothelial dysfunction rats. Applications of simvastatin and tongxinluo to endothelial dysfunction rats improved endothelial function according to the results of histopathology and measurements of endothelin-1 and nitric oxide. Metabonomic studies suggested that tongxinluo prevents endothelial dysfunction by regulating multiple metabolic pathways to their normal state, whereas simvastatin only altered selected metabolic pathways. This research demonstrated that metabonomics is a powerful and promising tool for disease investigation and the efficacy evaluation of complex traditional Chinese medicines. PMID- 21620605 TI - Targeting mitochondrial oxidative stress in heart failure throttling the afterburner. PMID- 21620606 TI - Mitochondrial targeted antioxidant Peptide ameliorates hypertensive cardiomyopathy. AB - OBJECTIVES: We investigated the effect of reducing mitochondrial oxidative stress by the mitochondrial-targeted antioxidant peptide SS-31 in hypertensive cardiomyopathy. BACKGROUND: Oxidative stress has been implicated in hypertensive cardiovascular diseases. Mitochondria and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase have been proposed as primary sites of reactive oxygen species (ROS) generation. METHODS: The mitochondrial targeted antioxidant peptide SS-31 was used to determine the role of mitochondrial oxidative stress in angiotensin II (Ang)-induced cardiomyopathy as well as in Galphaq overexpressing mice with heart failure. RESULTS: Ang induces mitochondrial ROS in neonatal cardiomyocytes, which is prevented by SS-31, but not the nontargeted antioxidant N-acetyl cysteine (NAC). Continuous administration of Ang for 4 weeks in mice significantly increased both systolic and diastolic blood pressure, and this was not affected by SS-31 treatment. Ang was associated with up-regulation of NADPH oxidase 4 (NOX4) expression and increased cardiac mitochondrial protein oxidative damage, and induced the signaling for mitochondrial biogenesis. Reducing mitochondrial ROS by SS-31 substantially attenuated Ang-induced NOX4 up regulation, mitochondrial oxidative damage, up-regulation of mitochondrial biogenesis, and phosphorylation of p38 mitogen-activated protein kinase and prevented apoptosis, concomitant with amelioration of Ang-induced cardiac hypertrophy, diastolic dysfunction, and fibrosis, despite the absence of blood pressure-lowering effect. The NAC did not show any beneficial effect. The SS-31 administration for 4 weeks also partially rescued the heart failure phenotype of Galphaq overexpressing mice. CONCLUSIONS: Mitochondrial targeted peptide SS-31 ameliorates cardiomyopathy resulting from prolonged Ang stimulation as well as Galphaq overexpression, suggesting its potential clinical application for target organ protection in hypertensive cardiovascular diseases. PMID- 21620607 TI - Humerus axial traction with acromial fixation reduction maneuver for anterior shoulder dislocation. AB - BACKGROUND: Many techniques have been described for the reduction of anterior glenohumeral dislocation, but each of the techniques has its disadvantages. A new shoulder reduction technique is needed to overcome these disadvantages. DESCRIPTION: An alternate technique of humerus axial traction with acromial fixation is presented. The technical description of this procedure focuses on the pre-reduction and post-reduction process. The use of acromial countertraction, the choice of the most loosely packed position of the shoulder joint, and the operator's ability to reduce muscle spasm are the main principles discussed. CONCLUSION: This modified technique increases the possibilities for the reduction of shoulder dislocation in different clinical situations. PMID- 21620608 TI - Consultation in the emergency department: a qualitative analysis and review. AB - BACKGROUND: No studies have evaluated the consultation process or attempted to define a standardized approach that could improve communication and patient outcomes. OBJECTIVE: To perform a qualitative analysis of emergency medicine (EM) consultation to reveal its complexity and elucidate strategies and frameworks for physician-to-physician communication. METHODS: Data were collected in three phases: informal interviews conducted in an emergency department (ED), 10 question surveys given to a subset of EM and specialty physicians, and semi structured 1-h group interviews using open-ended questions to further explore issues and trends elicited from the survey responses. In addition, we conducted an extensive literature search focused on health care and business consultation and communication. RESULTS: Seventy-six percent (29 of 38) of emergency and specialty physicians completed the 10-question survey in its entirety. Three themes were identified from the survey responses: organizational skills, interpersonal and communication skills, and medical knowledge. Of 95 total comments, 41 (43%) focused on organizational skills, 26 (27%) on interpersonal and communication skills, and 28 (30%) on medical knowledge. There were 29 comments regarding poor consultations: 15 issues with organization, 6 with interpersonal and communication skills, and 8 with medical knowledge. The literature search revealed several models and types of consultation, but no standard algorithm currently exists. CONCLUSIONS: We recommend focusing on organizational skills, interpersonal and communication skills, and medical knowledge when teaching ED consultation and present a conceptual framework of the Five Cs Consultation Model: contact, communication, core question, collaboration, and closing the loop. PMID- 21620609 TI - Stroke complicating traumatic ventricular septal defect. AB - BACKGROUND: Traumatic ventricular septal defect (VSD) occurs in approximately 5% of blunt or penetrating cardiac injuries and can result in rare complications. OBJECTIVES: To report the serious complication of stroke after a traumatic VSD. CASE REPORT: A 27-year-old man with no previous medical history presented to the Emergency Department with aphasia and right hemiparesis after a stab wound to the chest. He underwent emergent evacuation of a pericardial effusion and repair of a right ventricular wall perforation. Head computed tomography revealed left middle cerebral artery infarct. Post-operatively, he was noted to have a cardiac murmur, and echocardiogram revealed a VSD. The VSD was surgically repaired without complication. CONCLUSION: Stroke can complicate traumatic VSDs. PMID- 21620610 TI - Comparison of the diagnostic characteristics of two B-type natriuretic peptide point-of-care devices. AB - BACKGROUND: B-type natriuretic peptide (BNP) is used to diagnose heart failure (HF). OBJECTIVE: To compare the accuracy of two commercially available point-of care (POC) devices for measuring B-type natriuretic peptide (BNP) in emergency department (ED) patients with suspected heart failure using the central laboratory testing results as the criterion standard. METHODS: Venous blood samples were collected from adults with suspected heart failure and split into three samples for BNP analysis: central laboratory (Siemens ADIVA Centaur; Siemens, Deerfield, IL), Triage BNP POC device (Biosite, San Diego, CA), and i STAT BNP POC device (Abbott, East Windsor, NJ). The criterion standard for BNP levels was the central laboratory. RESULTS: Two hundred fifty patients were enrolled. Mean (SD) age was 70.7 (13.8) years; 200 (80%) were over age 55 years; 146 (58.4%) were male. A final hospital discharge diagnosis of heart failure was made in 108 (42%) patients. The i-STAT system yielded a result within a median of 9 min (interquartile range [IQR] 9-10 min). The Triage device yielded a result within a median of 19 min (IQR 15-22 min); p < 0.001. The device failure rate for the central laboratory (8 failures, 3.2%) was significantly higher than that of the i-STAT device (1 failure, 0.4%, p = 0.04), but not statistically different than the Triage device (3 failures, 1.2%). Neither the Triage nor the i-STAT were statistically different than the central laboratory result in terms of sensitivity; the i-STAT was less specific than the Triage result (p = 0.003). The area under the curve for the Triage device was 0.95 (95% confidence interval [CI] 0.91-0.98), whereas the area under the curve for the i-STAT device was 0.98 (95% CI 0.96-0.99; p < 0.01). CONCLUSIONS: Both POC devices tested were accurate and rarely failed; however, the i-STAT was faster with single use. PMID- 21620611 TI - Long-term results of open and endovascular revascularization of superficial femoral artery occlusive disease. AB - BACKGROUND: First-line treatment for patients with superficial femoral arterial (SFA) occlusive disease has yet to be determined. This study compared long-term outcomes between primary SFA stent placement and primary femoral-popliteal bypass. Periprocedural patient factors were examined to determine their effect on these results. METHODS: All femoral-popliteal bypasses and SFA interventions performed in consecutive patients with symptoms Rutherford 3 to 6 between 2001 and 2008 were reviewed. Time-dependent outcomes were analyzed using the Kaplan Meier method and log-rank test. Cox proportional hazards were performed to determine predictors of graft patency. Multivariate analysis was completed to identify patient covariates most often associated with the primary therapy. RESULTS: A total of 152 limbs in 141 patients (66% male; mean age, 66 +/- 22 years) underwent femoral-popliteal bypass, and 233 limbs in 204 patients (49% male; mean age, 70 +/- 11 years) underwent SFA interventions. Four-year primary, primary-assisted, and secondary patency rates were 69%, 78%, and 83%, respectively, for bypass patients and 66%, 91%, and 95%, respectively, for SFA interventions. Six-year limb salvage was 80% for bypass vs 92% for stenting (P = .04). Critical limb ischemia (CLI) and renal insufficiency were predictors of bypass failure. Claudication was a predictor of success for SFA stenting. Three year limb salvage rates for CLI patients undergoing surgery and SFA stenting were 83%. Amputation-free survival at 3 years for CLI patients was 55% for bypass and 59% for SFA interventions. Multivariate predictors (odds ratios and 95% confidence intervals) of covariates most frequently associated with first-line SFA stenting were TransAtlantic Inter-Society Consensus II A and B lesions (5.9 [3.4-9.1], P < .001), age >70 years (2.1 [1.4-3.1], P < .001), and claudication (1.7 [1.1-2.7], P = .01). Regarding bypass as first-line therapy, claudicant patients were more likely to have nondiabetic status (5.6 [3.3-9.4], P < .001), creatinine <1.8 mg/dL (4.6 [1.5-14.9], P = .01), age <70 years (2.7 [CI, 1.6 8.3], P < .001), and presence of an above-knee popliteal artery target vessel (1.9 [CI, 1.1-3.4] P = .02). CONCLUSION: Indication, patient-specific covariates, and anatomic lesion classification have significant association when determining surgeon selection of SFA stenting or femoral-popliteal bypass as first-line therapy. Patients with SFA disease can have comparable long-term results when treatment options are well matched to patient-specific and anatomic characteristics. PMID- 21620613 TI - Discussion. PMID- 21620612 TI - Efficacy of fragmin/protamine microparticles containing fibroblast growth factor 2 (F/P MPs/FGF-2) to induce collateral vessels in a rabbit model of hindlimb ischemia. AB - OBJECTIVES: The localized delivery of exogenous, angiogenic growth factors such as fibroblast growth factor (FGF)-2 has become a promising alternative treatment of peripheral artery disease (PAD) and critical limb ischemia (CLI). The present study describes the efficacy of fragmin/protamine microparticles containing FGF-2 (F/P-MPs/FGF-2) to promote vessel growth in a rabbit model of hindlimb ischemia. METHODS: A total of 24 rabbits were used to construct a model of hindlimb ischemia by resection of the left femoral artery. The rabbits were randomly divided into four groups 10 days after surgery (day 0); group A: control (non treated; 1 mL of phosphate-buffered saline [PBS]); group B: FGF-2 (100 MUg FGF-2 in 1 mL PBS)-treated; group C: F/P-MPs (12 mg dried F/P MPs in 1 mL PBS)-treated; and group D; F/P MPs/FGF-2 (100 MUg FGF-2 and 12 mg dried F/P MPs in 1 mL PBS) treated (n = 6 each). The drugs were administered intramuscularly to each group. Blood flow and blood pressure were measured in each group on days 0, 14, and 28. Angiography was performed to assess arteriogenesis on day 28. The number of capillaries on day 28 was determined by direct counting CD31(-) and alpha-smooth muscle antibody (alpha-SMA)-positive vessels. RESULTS: Neither death nor wound infection was observed throughout the experiment. The F/P MPs/FGF-2-treated group showed marked improvement in the blood flow ratio, blood pressure ratio, and capillary number in comparison to the control group, FGF-2-treated group, and F/P MPs-treated group. The F/P MPs-treated group showed intermediate improvement in blood flow ratio and capillary number in comparison to the control group and FGF 2-treated group. CONCLUSIONS: The F/P MPs/FGF-2-treated group strongly induced functional collateral vessels in the rabbit model of hindlimb ischemia, indicating a possible therapy for PAD. PMID- 21620614 TI - Duplex ultrasound screening detects high rates of deep vein thromboses in critically ill trauma patients. AB - OBJECTIVE: American College of Chest Physician (ACCP) guidelines stratify deep venous thrombosis (DVT) risk in trauma patients based on injury pattern and pharmacologic prophylaxis. Screening is only recommended for patients with high risk injuries who are unable to receive pharmacologic prophylaxis. However, the prevalence of lower extremity DVT (LEDVT) in trauma patients may be higher than reported in previous studies as many studies on DVT screening have not investigated calf vein DVTs (CVDVT) and have not exclusively targeted critically ill patients. Given that current ACCP guidelines recommend treatment of CVDVTs, we investigated the efficacy of duplex ultrasound (DUS) screening in critically ill trauma patients for all LEDVTs, including CVDVT, regardless of injury pattern, risk factors, or pharmacologic prophylaxis. METHODS: Medical records of 264 intensive care unit trauma patients who received DUS screening for LEDVT were retrospectively examined for the presence of injuries conferring high risk for LEDVT, patient specific DVT risk factors, and low molecular weight heparin (LMWH) prophylaxis. RESULTS: Forty (15.2%) patients had LEDVTs found on DUS screening, 24 (60%) were CVDVT, and 30% of all DVTs were diagnosed within 1 week of admission. Patients without high-risk injuries receiving LMWH had a 13.5% DVT rate, which did not differ significantly from the 19.7% DVT rate in high-risk injury patients not receiving LMWH (P = .667). CONCLUSIONS: Lower extremity DVT is common in critically ill trauma patients, particularly in the first week following injury, regardless of injury pattern, DVT risk factors, or pharmacologic prophylaxis. Previous studies have underestimated DVT rates by not investigating CVDVTs and not exclusively targeting critically ill patients. We recommend early and continued DUS DVT screening of all critically ill trauma patients. PMID- 21620615 TI - Resident and fellow experiences after the introduction of endovascular aneurysm repair for abdominal aortic aneurysm. AB - OBJECTIVES: This study assessed trends in open and endovascular repair (EVAR) of intact and ruptured abdominal aortic aneurysm (AAA) in the Medicare population and evaluated recent trends in AAA repair at vascular fellowship training programs. METHODS: We identified all Medicare beneficiaries with a diagnosis of AAA who underwent repair or had a primary diagnosis of rupture (1995-2008). Cohorts were compared by type of repair (open vs EVAR) and presentation (intact vs ruptured AAA). Demographics of age, sex, and race were evaluated. We used unique hospital identifier codes to compare trends and 30-day mortality between hospitals that participate in vascular surgery fellowship training and those that do not. American Council on Graduate Medical Education data, only available for the years 1999 to 2008, were further used to better understand the changes in number of EVAR and open repairs of AAA performed each year for vascular fellows and general surgery residents, over time. RESULTS: We identified 449,122 patients (76% men), with 376,355 intact AAAs (84%) and 72,767 ruptured AAAs (16%). Mean age was 75.1 years. Use of EVAR for intact AAA rose to from 35% in 2001 to 63% in 2005 and comprised 78% of repairs by 2008. During the same period, the number of ruptured AAAs decreased by 40% overall, with nonoperative ruptured AAAs decreasing by 29% and EVAR increasing to 31% of rupture repairs. Hospitals training vascular fellows were quicker to adopt EVAR (2-year lag time) for intact AAA and had higher rates of EVAR for ruptured AAA (41.1% vs 29.2%; P = .001) than did hospitals without fellows. Mortality rates for open repairs of intact (4.0% vs 5.0%; P = .01) and ruptured AAA (34.1% vs 41.0%; P = .031) were lower at fellowship hospitals. The average number of open AAA repairs performed by vascular fellows dropped 50% (44.1 to 21.6/year) from 1999 to 2008. CONCLUSIONS: Contrary to the expectation of a plateau, use of EVAR for intact AAA continues to rise at fellowship and nonfellowship hospitals. Use of EVAR for rupture is being used more often at fellowship programs. The decline in open repairs performed by vascular fellows, and at fellowship and non-fellowship hospitals, may have important implications for future attending experience. PMID- 21620617 TI - Inflow thrombosis does not adversely affect thrombolysis outcomes of symptomatic iliofemoral deep vein thrombosis. AB - OBJECTIVE: The presence of popliteal or tibial vein clot is thought to adversely affect thrombolysis for iliofemoral deep vein thrombosis (DVT). We examined the effect of inflow thrombosis on functional and anatomic outcomes. METHODS: Data for 44 patients treated for symptomatic iliofemoral DVT between 2006 and 2009 were retrospectively reviewed. All patients were treated by pharmacomechanical thrombectomy with local lytic therapy. Catheter-directed lysis and vena cava filters were used sparingly. Univariate and multivariate logistic regression analyses were used. The independent variable used in the logistic regression model was symptom relief. RESULTS: Forty-four patients (mean age, 52.1 +/- 15.8 years) presented with symptoms averaging 13.4 +/- 9.9 days in duration. Twenty (45.4%) had symptoms for >14 days. Seventeen patients were treated in one session, but 27 patients required lytic infusion for residual thrombus. Iliac stenting was required in 49% of limbs. Successful lysis (>50%) was achieved in 91% of patients, and symptom resolution or improvement in 91%. All patients became ambulatory, with no or minimal limitation. No major systemic bleeding complications occurred. Freedom from DVT recurrence and reintervention was 84% at 24 months by life-table analysis. Preoperative ultrasound imaging showed 89% had popliteal and tibial clots. A thrombosed popliteal vein was accessed for treatment and was corroborated by venographic findings. One patient required simultaneous tibial lysis. At a mean follow up of 8.7 +/- 6.3 months, 41 patients (93%) had no symptom recurrence, 82% had preserved valve function and no reflux on duplex imaging, with a mean CEAP class of 1.4 and Villalta score of 3.3. Inflow thrombus had no adverse effect on symptom relief, treatment duration, patency, CEAP class, or valve reflux. Interestingly, 90% of patients with initial popliteal thrombus had a patent popliteal vein on postlysis ultrasound imaging, and the presence of tibial thrombus on presentation was predictive of symptom relief with thrombolysis (odds ratio, 13.03; 95% confidence interval, 1.02 165.58; P = .048). CONCLUSIONS: Inflow thrombosis is common and does not preclude successful thrombolysis of iliofemoral DVT. Valve function is preserved on midterm follow-up, with maintained CEAP class and symptom relief. PMID- 21620618 TI - Reproductive history in women with abdominal aortic aneurysms. AB - BACKGROUND: The prevalence of abdominal aortic aneurysms (AAAs) differs considerably between the sexes, illustrated by the male/female ratio 4-6:1. Women are also reported to have a higher risk of rupture, and a poorer outcome compared with men. The primary aim of this study was to investigate if women with AAA have a different reproductive history compared with other women. The secondary aim was to study if women with a larger AAA differ in their reproductive history from women with a smaller AAA. METHOD: This case-control study was performed in October 2009 and included 140 consecutively monitored women with AAA and 140 with peripheral arterial disease (PAD) at the Department of Vascular Surgery at Karolinska University Hospital, Stockholm. AAA was defined as AAA diameter >3 cm, and women with AAA were subdivided into groups with AAA diameter >=5 cm and diameter <5 cm. A validated questionnaire was used to obtain information about participants' reproductive history and general health. The response rate was 70% (n = 196). RESULTS: Women with AAA were smokers to a greater extent than women with PAD (previous, 52% vs 46%; current, 46% vs 34%, P = .001). Diabetes mellitus was more prevalent in women with PAD (28%) than in women with AAA (15%, P = .034). Angina pectoris occurred more often in women with AAA (26%) than in women with PAD (11%, P = .026). No significant difference was found between PAD and AAA women regarding statin use, treatment for hypertension, prior myocardial infarction, and body mass index (BMI). The 54 women with AAA >=5 cm and the 44 women with AAA <5 cm were similar in age (76 vs 76 years, P = .908) and BMI (25.7 vs 24.0 kg/m(2), P = .66). No difference was noted in the occurrence of other risk factors between women with AAA >=5 cm and women with AAA <5 cm. Mean age at menopause was lower in women with AAA >=5 cm than in women with AAA <5 cm and in women with PAD (47.7 vs 49.9 vs 49.7 years, P = .011). Apart from menopausal age, the groups had a similar reproductive history, including hormone replacement therapy, parity, use of contraceptives, prior gynecological surgery, and breast cancer. CONCLUSION: These findings suggest that women with larger AAA reach menopausal age earlier, and this could influence an earlier onset of aneurysmatic disease or an increase in aneurysm growth. The true role of endogenous estrogen in aneurysm development and expansion is yet to be investigated. PMID- 21620619 TI - Comparison of modern open infrarenal and pararenal abdominal aortic aneurysm repair on early outcomes and renal dysfunction at one year. AB - OBJECTIVE: This study was conducted to review contemporary results of elective open infrarenal abdominal aortic aneurysm (IAAA) and pararenal abdominal aortic aneurysm (PAAA) repairs and determine predictors of death and acute and 1-year renal dysfunction (RD). METHODS: A retrospective review identified 432 consecutive patients undergoing open IAAA (233 patients) or PAAA (184 patients) repair between January 2000 and December 2007. Demographic, preoperative, intraoperative, and postoperative variables were collected. RD was defined as an increase in creatinine of >= 0.5 mg/dL from baseline. Multiple logistic regression models were used to identify predictors of mortality and RD. RESULTS: Mortality rates were similar between the groups (3.9% IAAA and 6.0% PAAA). Preoperative coronary artery disease (CAD), postoperative myocardial infarction, or pulmonary complications were all strong predictors of operative mortality in patients undergoing repair of PAAAs and IAAAs. However, neither PAAA nor baseline renal insufficiency was an independent predictor of death. Postoperative RD occurred in 32% of patients after PAAA repairs compared with 13% of patients after IAAA repairs (P < .001). The presence of PAAA, baseline hypertension, and hyperlipidemia all correlated positively with postoperative RD, while a trend was noted with baseline renal insufficiency (P = .09). At the 1-year follow-up, 5.1% of patients in the PAAA group had RD compared with none in the IAAA group. Similarly, the serum creatinine level was significantly higher in the PAAA group (1.4 mg/dL vs 1.2 mg/dL, PAAA and IAAA, respectively; P = .02) at 1 year. However, there were no instances of new-onset hemodialysis dependence at 1 year. Mean follow-up was 2.2 years overall. CONCLUSION: Open PAAA repair can be performed without a significant increase in mortality compared to open IAAA repair. Although the incidence of renal function deterioration after open PAAA repairs remains higher than with open IAAA repairs, the overall incidence remains low at 1-year follow-up. PMID- 21620620 TI - Endovascular treatment of thoracic aorta injury after spinal column surgery. PMID- 21620621 TI - WITHDRAWN: A novel approach using pulmonary artery catheter-directed rapid right ventricular pacing to facilitate precise deployment of endografts in the thoracic aorta. AB - The Publisher regrets that this article is an accidental duplication of an article that has already been published, doi: 10.1016/j.jvs.2011.10.003. The duplicate article has therefore been withdrawn. PMID- 21620623 TI - Evaluation of robotic endovascular catheters for arch vessel cannulation. AB - OBJECTIVE: Conventional catheter instability and embolization risk limits the adoption of endovascular therapy in patients with challenging arch anatomy. This study investigated whether arch vessel cannulation can be enhanced by a remotely steerable robotic catheter system. METHODS: Seventeen clinicians with varying endovascular experience cannulated all arch vessels within two computed tomography-reconstructed pulsatile flow phantoms (bovine type I and type III aortic arches), under fluoroscopic guidance, using conventional and robotic techniques. Quantitative (catheterization times, catheter tip movements, vessel wall hits, catheter deflection) and qualitative metrics (Imperial College Complex Endovascular Cannulation Scoring Tool [IC3ST]) performance scores were compared. RESULTS: Robotic catheterization techniques resulted in a significant reduction in median carotid artery cannulation times and the median number of catheter tip movements for all vessels. Vessel wall contact with the aortic arch wall was reduced to a median of zero with robotic catheters. During stiff guidewire exchanges, robotic catheters maintained stability with zero deflection, independent of the distance the catheter was introduced into the carotid vessels. Overall IC3ST performance scores (interquartile range) were significantly improved using the robotic system: Type I arch score was 26/35 (20-30.8) vs 33/35 (31-34; P = .001), and type III arch score was 20.5/35 (16.5-28.5) vs 26.5/35 (23.5-28.8; P = .001). Low- and medium-volume interventionalists demonstrated an improvement in performance with robotic cannulation techniques. The high-volume intervention group did not show statistically significant improvement, but cannulation times, movements, and vessel wall hits were significantly reduced. CONCLUSION: Robotic technology has the potential to reduce the time, risk of embolization and catheter dislodgement, radiation exposure, and the manual skill required for carotid and arch vessel cannulation, while improving overall performance scores. PMID- 21620624 TI - Endovascular aneurysm repair reverses the increased titer and the inflammatory activity of interleukin-1alpha in the serum of patients with abdominal aortic aneurysm. AB - OBJECTIVE: To examine serum cytokine/chemokine profiles before and 6 months after endovascular repair (EVAR) of abdominal aortic aneurysm (AAA) and to determine whether they correlate with serum inflammatory activity using an in vitro model of leukocyte recruitment. METHODS: Serum IL-1-alpha, IL-1beta, IL-4, IL-6, IL-8, IL-10, IFN-gamma, IP-10, MCP-1, TNF-alpha, and TNF-beta were measured using a cytometry-based immunoassay. To test patient serum for direct inflammatory activity, human endothelial cells (EC) were stimulated with 30% patient serum for 24 hours. To test patient serum for the ability to prime EC for inflammatory responses, EC were incubated with 30% patient serum for 24 hours, followed by stimulation with low-dose (5 U/mL) TNF for 4 hours. Under both regimens of stimulation, the degree of EC activation was assessed by assaying neutrophil recruitment in a flow-based model. RESULTS: Only IL-1alpha (67.9 +/- 10.4 pg/mL vs 41.9 +/- 7.4 pg/mL) and IL-8 (51.5 +/- 5.1 vs 32.6 +/- 4.7 pg/mL) changed significantly after surgery. Patient serum alone was unable to activate EC. However, serum from both time points could prime EC responses to low-dose TNF. Thus, after priming with preoperative serum, EC stimulated with TNF could recruit 76.7 +/- 12.0 neutrophils/mm(2) into the subendothelial cell space. Post-EVAR serum was significantly less effective (44.4 +/- 10.2 neutrophils/mm(2)). This reduction in neutrophil recruitment correlated with reduced IL-1alpha in post EVAR serum. The addition of a neutralizing antibody against IL-1alpha to pre-EVAR serum inhibited EC priming and neutrophil recruitment, strongly implying that this cytokine was the priming agent. CONCLUSION: EVAR reduces serum IL-1alpha and its inflammatory activity in patient serum. IL-1alpha is, therefore, implicated in the molecular pathology of AAAs and may have potential as a clinically useful biomarker. PMID- 21620625 TI - Serum metalloproteinases MMP-2, MMP-9, and metalloproteinase tissue inhibitors in patients are associated with arteriovenous fistula maturation. AB - OBJECTIVE: Many vascular surgeons construct arteriovenous fistulas (AVFs) for hemodialysis access as the primary choice access. A significant number of AVFs fail to mature, however, leading to patient frustration and repeated operations. Metalloproteinase (MMP) activity, particularly MMP-2 and MMP-9, may be important for AVF maturation. We therefore sought to identify whether serum MMP levels could serve as a biomarker for predicting future successful AVF maturation. METHODS: Blood was collected from patients with chronic renal insufficiency at the time of surgery for long-term hemodialysis access. Serum was separated from whole blood and ultracentrifuged at 1000g for 10 minutes. Serum aliquots were frozen at -80 degrees C until used for analysis. Enzyme-linked immunosorbent assay was used to assay levels of MMP-2, MMP-9, and tissue inhibitor of metalloproteinase type 2 (TIMP-2), and TIMP type 4 (TIMP-4). Clinical end points were used to divide patients into failed and matured AVF groups. Successful maturation was considered in patients who had specific duplex findings or 1 month of successful two-needle cannulation hemodialysis. MMP/TIMP ratios were calculated as an index of the MMP axis activity because MMP activity parallels alterations in TIMP levels. RESULTS: Of 20 enrolled patients, AVF maturation was successful in 13 and failed in 7. Serum levels of MMP-2/TIMP-2 were significantly higher in patients with matured AVFs vs levels in those that failed (P = .003). Similarly, a trend toward increased serum levels of MMP-9/TIMP-4 was found in patients with successful AVF (P = .06). CONCLUSIONS: MMP-2 and TIMP-2 levels were different among patients whose AVF matured vs those who did not. Further follow up studies to determine the predictability of AVF maturation using relative patient serum levels of MMP-2 and TIMP-2 should be performed. PMID- 21620626 TI - Long-term outcomes of internal carotid artery dissection. AB - OBJECTIVE: The natural history of acute carotid artery dissection is poorly characterized. The purpose of this study is to report on single institutional long-term outcomes. METHODS: A retrospective review of patients treated for acute spontaneous or posttraumatic carotid artery dissection over a 20-year period from August 1989 to July 2009 was performed. RESULTS: Twenty-nine patients with a mean age of 47 +/- 19.6 years were identified with acute carotid dissection. Six (25%) were related to trauma, while 23 (79%) were spontaneous. Neurologic symptoms included contralateral limb weakness (55%), facial pain (35%), and Horner's syndrome (21%). Eight patients (28%) presented with an acute hemispheric stroke. Diagnostic imaging modalities used included computed tomography angiography (52%), magnetic resonance angiography (41%), and conventional angiography (48%). Twenty percent of patients had complete carotid occlusion and 25% had near occlusion. Most dissections (65%) had intracranial extension, and 35% were limited to the extracranial cervical internal carotid. The majority (96%) of patients were treated conservatively with anticoagulation or antiplatelet therapy or both. One patient underwent stenting for persistent symptoms resulting in complete recovery. There were two deaths, one from unrelated traumatic injuries and the other from unknown causes. Long-term follow-up was available for 20 patients: 14 had complete symptom resolution (70%) and five (25%) had partial clinical symptom resolution. Two patients had initial resolution of symptoms, with subsequent recurrence that was successfully managed conservatively. Follow up imaging revealed luminal patency in 79% of patients with minimal residual stenosis. Two patients developed a small asymptomatic internal carotid aneurysm that did not require treatment. Mean follow-up was 1133.2 days. CONCLUSIONS: Most cervical carotid dissections can safely be conservatively managed, with the majority achieving anatomic and symptomatic resolution, with low rates of recurrence over long-term follow-up. PMID- 21620627 TI - In vivo three-dimensional blood velocity profile shapes in the human common, internal, and external carotid arteries. AB - OBJECTIVE: True understanding of carotid bifurcation pathophysiology requires a detailed knowledge of the hemodynamic conditions within the arteries. Data on carotid artery hemodynamics are usually based on simplified, computer-based, or in vitro experimental models, most of which assume that the velocity profiles are axially symmetric away from the carotid bulb. Modeling accuracy and, more importantly, our understanding of the pathophysiology of carotid bifurcation disease could be considerably improved by more precise knowledge of the in vivo flow properties within the human carotid artery. The purpose of this work was to determine the three-dimensional pulsatile velocity profiles of human carotid arteries. METHODS: Flow velocities were measured over the cardiac cycle using duplex ultrasonography, before and after endarterectomy, in the surgically exposed common (CCA), internal (ICA), and external (ECA) carotid arteries (n = 16) proximal and distal to the stenosis/endarterectomy zone. These measurements were linked to a standardized grid across the flow lumina of the CCA, ICA, and ECA. The individual velocities were then used to build mean three-dimensional pulsatile velocity profiles for each of the carotid artery branches. RESULTS: Pulsatile velocity profiles in all arteries were asymmetric about the arterial centerline. Posterior velocities were higher than anterior velocities in all arteries. In the CCA and ECA, velocities were higher laterally, while in the ICA, velocities were higher medially. Pre- and postendarterectomy velocity profiles were significantly different. After endarterectomy, velocity values increased in the common and internal and decreased in the external carotid artery. CONCLUSIONS: The in vivo hemodynamics of the human carotid artery are different from those used in most current computer-based and in vitro models. The new information on three-dimensional blood velocity profiles can be used to design models that more closely replicate the actual hemodynamic conditions within the carotid bifurcation. Such models can be used to further improve our understanding of the pathophysiologic processes leading to stroke and for the rational design of medical and interventional therapies. PMID- 21620628 TI - Frequency, risk factors, and management of perigraft seroma after open abdominal aortic aneurysm repair. AB - OBJECTIVE: Perigraft seroma (PGS) causing enlargement of the native aneurysm sac after open abdominal aortoiliac aneurysm (AAA) repair is a rarely recognized complication with unknown clinical consequences. This study was undertaken to determine the frequency of PGS, identify associated risk factors, and review resulting complications and their management strategies. METHODS: Charts of all patients who underwent open AAA repair at our institution from 1995 to 2009 and had at least one postoperative abdominal cross-sectional imaging study (the study subjects) were retrospectively reviewed. PGS was defined as a perigraft fluid collection present > 3 months postoperatively, >= 3-cm in diameter and having a radiodensity <= 25 Hounsfield units on computed tomography (CT). Patient records were reviewed for demographics, comorbidities, operative and postoperative variables, and long-term outcome. RESULTS: Of the 111 study subjects identified, 13 had aortic reconstruction with Dacron grafts and 98 with polytetrafluoroethylene (PTFE) grafts. Twenty patients (18%) had PGS, all of whom had PTFE grafts (20 of 98; 20.4%). Mean age was 68.5 years and mean aneurysm diameter preoperatively was 6.4 cm (range, 4.0-10.9 cm). The average time from AAA repair to PGS detection was 51 months (range, 4-156 months). PGS averaged 6.0 cm in diameter (range, 3.0-11.0 cm). Multivariate analysis revealed that the following factors were associated with PGS development: diabetes (odds ratio [OR], 3.5; 95% confidence interval [CI], 1.1-21.2; P = .013), smoking (OR, 5.6; 95% CI, 0.73-33.74; P = .01), anticoagulation (OR, 7.2; 95% CI, 2.6-63.3; P = .003), bifurcated graft reconstruction (OR, 8.0; 95% CI, 2.6-94.1; P = .017), and left flank retroperitoneal approach for repair (OR, 7.1; 95% CI, 1.9-26.5; P = .003). Four patients (4 of 20; 20%) required intervention for PGS-related complications: 3 patients for symptomatic PGS expansion (1 patient with rupture) and 1 patient for acute limb ischemia secondary to graft limb compression and thrombosis. Two patients had open exploration, sac evacuation/reduction, and graft replacement with a Dacron graft: 1 patient for a ruptured aneurysm sac and 1 patient for persistent pain associated with sac enlargement. A third patient underwent a failed CT-guided drainage for abdominal pain and was subsequently treated with partial graft excision. The patient with acute limb ischemia was treated with catheter-directed thrombolysis and graft limb stenting. CONCLUSION: PGS after open AAA repair occurs more frequently than previously reported. Complications requiring intervention can occur in up to 20% of patients with PGS. A variety of treatment modalities can be used to deal with the complications. Earlier CT surveillance is advised after open AAA repair with a PTFE graft if symptoms are suggestive of PGS development. PMID- 21620629 TI - Intravenous leiomyomatosis presenting as acute Budd-Chiari syndrome. AB - Intravenous leiomyomatosis is a rare condition that has been described as being associated with venous occlusion by direct intravascular tumor extension, typically from a pelvic organ. We report an exceedingly rare case of intravenous leiomyomatosis extending into the inferior vena cava, leading to pulmonary embolism, hepatic venous outflow obstruction, and an acute Budd-Chiari syndrome. This is the second reported patient with intravenous leiomyomatosis with Budd Chiari syndrome and, to our knowledge, the first reported patient who survived with surgery. Correlative images, illustrating computed tomography and magnetic resonance imaging findings characteristic of intravenous leiomyomatosis with secondary Budd-Chiari syndrome, are presented and discussed. PMID- 21620630 TI - Cost-effectiveness of intensive smoking cessation therapy among patients with small abdominal aortic aneurysms. AB - INTRODUCTION: Smoking cessation is one of the few available strategies to decrease the risk for expansion and rupture of small abdominal aortic aneurysms (AAAs). The cost-effectiveness of an intensive smoking cessation therapy in patients with small AAAs identified at screening was evaluated. METHODS: A Markov cohort simulation model was used to compare an 8-week smoking cessation intervention with adjuvant pharmacotherapy and annual revisits vs nonintervention among 65-year-old male smokers with a small AAA identified at screening. The smoking cessation rate was tested in one-way sensitivity analyses in the intervention group (range, 22%-57%) and in the nonintervention group (range, 3% 30%). Literature data on the effect of smoking on AAA expansion and rupture was factored into the model. RESULTS: The intervention was cost-effective in all tested scenarios and sensitivity analyses. The smoking cessation intervention was cost-effective due to a decreased need for AAA repair and decreased rupture rate even when disregarding the positive effects of smoking cessation on long-term survival. The incremental cost/effectiveness ratio reached the willingness-to-pay threshold value of ?25,000 per life-year gained when assuming an intervention cost of > ?3250 or an effect of <= 1% difference in long-term smoking cessation between the intervention and nonintervention groups. Smoking cessation resulted in a relative risk reduction for elective AAA repair by 9% and for rupture by 38% over 10 years of follow-up. CONCLUSIONS: An adequate smoking cessation intervention in patients with small AAAs identified at screening can cost effectively increase long-term survival and decrease the need for AAA repair. PMID- 21620631 TI - One-year multicenter results of 100 abdominal aortic aneurysm patients treated with the Endurant stent graft. AB - OBJECTIVE: The Endurant (Medtronic, Minneapolis, Minn) is a new stent graft specifically designed to make more patients anatomically eligible for endovascular aneurysm (EVAR). This study presents the 1-year results of 100 consecutive patients with abdominal aortic aneurysms (AAAs) treated with the Endurant stent graft in real-life practice. METHODS: All clinical preoperative, operative, postoperative, and 1-year follow-up data of patients with the Endurant stent graft from three tertiary centers were prospectively collected. Patients underwent computed tomographic angiography (CTA) preoperatively, at 1 month, and at 1-year post-EVAR. The first 100 patients with an implantation date at least 1 year before our date of analysis and complete information were included. Clinical data, AAA characteristics, presence of endoleaks, graft migration, and other EVAR related complications were noted. All values are stated as mean +/- SD (range). RESULTS: This study included 100 patients with AAAs (88 men) with a mean age of 73 +/- 8 years (47 to 87 years), an AAA size of 61 +/- 10 mm (31 to 93 mm), an AAA volume of 210 +/- 122 mL (69 to 934 mL), a proximal neck length of 33 +/- 14 mm (9 to 82 mm), and an infrarenal angulation of 44 +/- 25 degrees (0 degrees 108 degrees ). Nineteen of the 100 included patients had at least one anatomic characteristic that was considered a violation of the instructions for use (IFU) of the Endurant stent graft. A primary technical success was achieved in 98% of the patients (one additional stent placement in renal artery was required; one unplanned aorto-uni-iliac device placed), with no primary type I or III endoleaks or conversions. A secondary technical success was achieved in all cases. The 30 day mortality was 2% and the first postoperative CTA documented 16 endoleaks (16%; 16 type II). One-year follow-up showed three iliac limb occlusions (3%), one infected stent graft (causing a type Ia endoleak), and five endovascular reinterventions (5%; three to treat iliac limb occlusions, one proximal extension cuff; and one stent in the renal artery). The 1-year all-cause mortality rate was 12% (12 patients) and the AAA-related mortality was 3%. The mean AAA size was significantly smaller after 1 year (diameter, 54 +/- 11.8 [32-80] mm; P < .01; volume, 173 +/- 119 [42-1028] mL; P < .01), and one graft migration >5 mm and 13 endoleaks were noted (12 type II, 1 type I [neck dilatation]). CONCLUSION: The treatment of patients with AAAs with the Endurant stent graft seems to be successful and durable during the first year after EVAR. Despite the wider inclusion criteria for the Endurant, and with 19% of our patients treated outside the IFU, the AAA-related mortality, number of type I or III endoleaks, and reintervention rates are comparable to the results of other stent grafts. PMID- 21620632 TI - Type IV collagen as a tumour marker for colorectal liver metastases. AB - BACKGROUND: About 50% of patients with primary colorectal cancer (CRC) will develop liver metastases (CLM). Currently, carcinoembryonic antigen (CEA) is the most common tumour marker for CRC and CLM. However, the sensitivity and specificity of this marker is not optimal, as almost 50% of patients have tumours that do not produce CEA. Therefore there is a need for better markers for CRC and CLM. METHODS: The circulating levels of type IV collagen were measured in patients with CLM, primary CRC and in healthy controls. The expression pattern of type IV collagen was studied by immunofluorescence in CLM and normal liver tissue. The metastatic volume of CLM in the liver was estimated from CT. RESULTS: In CLM tissue type IV collagen is highly expressed in the areas of desmoplasia. Patients with primary CRC (Dukes' A-C) did not show any increase in circulating type IV collagen compared to healthy controls. However, patients with CLM have significantly elevated levels of circulating type IV collagen when compared to patients with primary CRC and healthy controls. The levels of type IV collagen decreased during chemotherapy and increased at the time of disease progression. The circulating levels of type IV collagen seem to reflect the tumour burden in the liver. CONCLUSIONS: Type IV collagen has the potential to be used as tumour associated biomarker for CLM. These results indicate the importance of interaction between cancer cells and the stroma in the tumour microenvironment. PMID- 21620633 TI - The effect of intra-articular autogenous bone marrow injection on healing of an acute posterior cruciate ligament injury in rabbits. AB - PURPOSE: The purpose was to identify the effect of intra-articular autogenous bone marrow injection on the healing of an acute posterior cruciate ligament (PCL) rupture in a rabbit model. The effect of autogenous bone marrow on the healing process was assessed serially by gross inspection, histologic examination, and immunohistochemical study of growth factors. METHODS: In both knee joints, the PCL was completely transected surgically near the femoral attachment site in 24 rabbits. Autogenous bone marrow was obtained from both tibias and delivered only to the right knee joint by direct intra-articular injection. Gross inspection, histologic examination, and immunohistochemical study of growth factors were performed at 8, 12, and 16 weeks after severing of the posterior cruciate ligament (PCL) among 8 randomly chosen specimens. The degree of healing in both marrow-stimulated and untreated control ligaments was evaluated by gross inspection using an ordinal scale consisting of 5 grades. The degree of fibroblast and vessel proliferation and alignment of collagen fibers were evaluated by histologic examination. The degree of expression of transforming growth factor beta1, epidermal growth factor receptor, and vascular endothelial growth factor was evaluated by immunohistochemical study. Statistical analysis was performed with the Mann-Whitney U test. RESULTS: In the group with marrow-stimulated ligaments, the degree of healing was higher at 8 and 12 weeks by gross examination, whereas there was no significant difference at 16 weeks between the 2 groups. According to histologic examination, the healing process was faster in the bone marrow injection group than in the control group at 8 and 12 weeks because the degree of fibroblast and vessel proliferation significantly declined and collagen fibers were arranged more regularly compared with the control group. Similar to the results of histologic examination, the results of immunohistochemical studies showed that the healing process was faster in the bone marrow injection group. However, the recovery of the PCL was completed at 16 weeks after PCL resection in both the bone marrow injection group and the control group. CONCLUSIONS: Intra-articular autogenous bone marrow injection appeared to promote the initiation of healing response in acutely injured PCLs in rabbits. CLINICAL RELEVANCE: Intra-articular autogenous bone marrow injection can be a viable option for treating acutely injured PCLs. PMID- 21620634 TI - The posteromedial knee arthroscopy portal: a cadaveric study defining a safety zone for portal placement. AB - PURPOSE: To define a neurovascular safety zone for proper placement of at least 2 posteromedial (PM) portals and to describe a safe intra-articular entrance point based on the location of the PM capsular folds. METHODS: Arthroscopy was performed on 10 fresh-frozen cadaveric knees. With the knee flexed 90 degrees , PM portals were created under direct visualization with a 70 degrees arthroscope. The anatomic soft spot between the PM edges of the femoral condyle and the proximal tibia was palpated, and 18-gauge spinal needles were inserted into the joint followed by 5.0-mm cannulas. The PM capsular folds were used to guide portal locations. Four portals (A, B, C, and D) were created in each of the first 3 knees. Portal position was described in relation to the soft spot: through the soft spot (A), 1 cm superior (B), 1 cm posterior (C), and 1 cm inferior (D). Dissection was performed along the path of the cannulas. RESULTS: In the first 3 knees, the most inferior portal (D) always overlapped the course of the saphenous nerve, and the posterior portal (C) always pierced the gastrocnemius musculotendinous junction. Therefore these portals were eliminated from further study for safety reasons, and in the final 7 knees, we studied only the remaining 2 portal placements (A and B). Portals A and B were found to be the safest, with at least 1.5 cm of clearance between the portals and the saphenous nerve in all specimens. CONCLUSIONS: At least 2 PM portals can be safely placed in the knee. The soft spot is an appropriate landmark to ensure safe portal entry. The PM capsular folds can help guide intra-articular placement so that damage to surrounding structures can be avoided. CLINICAL RELEVANCE: We believe that 2 portals would be beneficial when performing complex arthroscopic procedures involving the PM compartment of the knee. PMID- 21620635 TI - Long-term follow-up of arthroscopic rotator cuff repair. AB - PURPOSE: The purpose of this study was to perform a case series assessing the clinical outcomes of patients with at least 9 years of follow-up after an all arthroscopic rotator cuff repair. METHOD: We performed a review of all of the arthroscopic rotator cuff repairs done by the senior author from 1991 to 2001. Study patients identified were contacted and evaluated by the first author and the senior author. A thorough in-office shoulder examination was completed and a current University of California, Los Angeles shoulder score was obtained during the evaluation. RESULTS: Seven hundred seventy-two patients were in the initial database. Forty-eight patients were identified from the database after inclusion and exclusion criteria were applied. Follow-up ranged from 110 to 223 months, averaging 151.7 months. All repairs were single row and received an arthroscopic subacromial decompression. We identified 33 all-arthroscopic rotator cuff repairs for follow-up in 24 patients included in the study. The mean University of California, Los Angeles score at follow-up was 31.8, with 87.7% of patients having excellent and good outcomes. Of the patients, 18 showed excellent results, 11 good, 2 fair, and 2 poor. All the patients presented with no loss of motion. CONCLUSIONS: Our data suggest that patients maintain good outcomes 10 years after the index surgery. These findings are comparable to the outcomes reported in short-term and midterm follow-up studies. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 21620636 TI - Femoral peel-off lesions in acute posterolateral corner injuries: incidence, classification, and clinical characteristics. AB - PURPOSE: The purpose of this study was to determine the incidence of acute avulsions of the popliteus tendon (PT) and lateral collateral ligament (LCL) of the femur (peel-off lesions) in knees with multiple injured ligaments. This study also describes the tear patterns and associations based on preoperative magnetic resonance imaging (MRI), physical examination, and arthroscopic inspection with injury patterns. METHODS: This was a retrospective review of a consecutive series of 48 acute grade III posterolateral knee repairs and reconstructions. All office evaluations were reviewed to determine the preoperative MRI examination results, the findings on physical examination with the patient under anesthesia, and the intraoperative arthroscopic findings. RESULTS: In total, 19 patients (40%) were surgically verified as having peel-off lesions. Tears were typically categorized into 3 patterns: isolated PT tear (pattern 1) (4 of 19 [21%]), combined PT and LCL tears (pattern 2) (8 of 19 [42%]), and complex tears (associated intrasubstance- and/or fibula-based injuries) (pattern 3) (7 of 19 [37%]). Of the patients, 82% (14 of 17) showed conclusive MRI signs of femoral insertion separation and discontinuity. Examination with the patient under anesthesia showed tibial external rotation of greater than 10 degrees in 84% of patients (16 of 19) and positive varus instability in 73% (11 of 15). Arthroscopic findings of acute avulsion and a positive lateral gutter drive-through sign were found in 94% (16 of 17). CONCLUSIONS: In a series of 48 grade III, consecutive, acute posterolateral corner surgery cases, we found that 19 patients (40%) had peel-off lesions. For adequate clinical identification, we recommend a comprehensive diagnostic protocol, including MRI, physical examination, and arthroscopic inspection, to avoid underdiagnosis of the lesion in multiple ligament-injured knees. The lateral gutter arthroscopic evaluation is particularly sensitive for detecting the lesion. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 21620637 TI - Effects of beach-chair position and induced hypotension on cerebral oxygen saturation in patients undergoing arthroscopic shoulder surgery. AB - PURPOSE: We investigated the effects of the beach-chair position and induced hypotension on regional cerebral oxygen saturation (rSO(2)) in patients undergoing arthroscopic shoulder surgery by using near-infrared spectroscopy. METHODS: Twenty-eight patients scheduled for arthroscopic shoulder surgery were enrolled prospectively. After induction of anesthesia, mechanical ventilation was controlled to maintain Paco(2) at 35 to 40 mm Hg. Anesthesia was maintained with sevoflurane and remifentanil. After radial artery cannulation, mean arterial pressure (MAP) was measured at the external auditory meatus level and maintained between 60 and 65 mm Hg. The rSO(2) was measured by use of near-infrared spectroscopy. MAP and rSO(2) were recorded at the following times: before induction (T(0)), immediately after induction (T(1) [baseline]), after beach chair position (T(2)), immediately after induced hypotension (T(3)), 1 hour after induced hypotension (T(4)), and after supine position at the end of surgery (T(5)). Cerebral desaturation was defined as a reduction in rSO(2) to less than 80% of baseline value for 15 seconds or greater. RESULTS: A total of 27 patients were evaluated until the end of this study. The MAP at T(2) was significantly lower than that at T(1). The MAP values at T(3) and T(4) were significantly lower than those at T(1) and T(2). The rSO(2) at T(2) was significantly lower than that at T(1). Unlike the pattern of change in the MAP, there was no additional decrease in rSO(2) at T(3) and T(4). There were 2 patients who had an episode of cerebral desaturation. CONCLUSIONS: The beach-chair position combined with induced hypotension significantly decreases rSO(2) in patients undergoing shoulder arthroscopic surgery under general anesthesia. LEVEL OF EVIDENCE: Level IV, study of nonconsecutive patients without consistently applied reference gold standard. PMID- 21620638 TI - [Observatory of the elderly over 80 years supported by the mobile emergency and resuscitation service]. AB - Emergency medical services (EMS) received an increasing number of calls for patients aged 80 and older. The goal of the present study was to evaluate outcome and functional dependence of patients aged 80 and older who EMS managed in the prehospital theater. This prospective study was conducted over 1 year (September 2007-August 2008), all consecutive patients aged 80 and older managed by a medical team during the study period were included. Characteristics of patients, including previous health status and functional dependence, were recorded on-the scene by the attending physician. Three-month mortality was recorded, as well as ADL score. Data are expressed as mean values+/-standard deviations, medians and interquartile ranges (IQRs), and percentages and compared using univariate and multivariate analysis. P<0.05 was considered the threshold for significance. Five hundred twenty-three patients were included. Mean age was 86 +/- 5. Median ADL index was 2 (IQR 0-9), and 63% of patients were living at home. At 3 months, the survival rate was 66% (n=273) and the proportion of patients living at home was 64% (P=0.9), the median ADL index of survivors was 2 (IQR 0-8) vs 1 (IQR 0-6) initially for this subpopulation, P=0.01. Our study confirms utility and efficacy of full access of elderly persons to advanced life support especially for self patients and not restricted based on aging per se. The development and daily use of tools for rapid assessment of autonomy should enable practitioners to innovate and thus, adapt their management. PMID- 21620639 TI - [Implementation strategy of the HAS French surgical check-list in a university hospital]. AB - The check-list (CL) "Safety in Operating Room" has been introduced in our teaching hospital since 2009, associated to a "Quality and Prevention of Risks" program. This introduction was carried out over two distinct phases. The first one was a pilot start including five OR, allowing us to draw firm recommendations on the best way to perform the introduction, followed by a generalization to the other operating room (OR). The recommendations were the followings: a pilot committee including all the professionals should be constituted before the onset of introduction, a dedicated communication should focus on the actual concerns and benefits, and finally, the person questioning other care givers and filling the form should be clearly identified and supported in the OR. Meanwhile a guide on the utilization of the CL in each surgical speciality was written, and a dedicated manager was in charge of the whole procedure. This experience raised several remarks. This implementation of the CL proved to be a cause of self interrogation on our medical practices, and the opportunity to improve communication among the professionals of the OR. Indeed, the 10 items of the OR should be thought as the last check before the no-return point, which should be shared by anyone in the OR. If these conditions were fulfilled, the CL could be viewed as an actual improvement of safety in the OR. Otherwise, CL is just a supplementary form. PMID- 21620640 TI - Susceptibility of Trypanosoma evansi to cordycepin. AB - Drugs, which are effective during the early stage of trypanosomosis, but poorly penetrate the blood-brain barrier, are ineffective when parasites reach the brain and cause encephalitis. In order to seek alternative treatments, the aim of this study was to test the susceptibility of T. evansi to cordycepin in vitro and in rats experimentally infected. In vitro, a significant decrease (P<0.01) in live trypanosomes in the concentrations of 5.0 and 10 MUg/mL was observed 1 hour after the beginning of the study, as well as at 3, 6, 9 and 12 hours in all concentrations compared to control. Although no curative effects were observed in the in vivo assay in the majority of groups, the drug was able to maintain parasitemia at low levels, therefore increasing the longevity of rats when compared to positive control group. Rats that received cordycepin alone or in combination with adenosine deaminase inhibitor (ADA: EHNA hydrochloride), did not show trypomastigote forms of the parasite in the bloodstream 24 hours after the administration. These animals remained negative in blood smears on average for 8 days, but thereafter had a recurrence of parasitemia. Among all the infected animals, only three rats in the group treated with the combination of cordycepin (2 mg/kg) and EHNA hydrochloride (2 mg/kg) remained negative during the experimental period. The curative efficacy of 42.5% was confirmed by PCR using T. evansi-specific primers. Thus, we conclude that cordycepin has biological effect against T. evansi, as previously reported in infections by T. brucei, T. cruzi and Leishmania sp. The treatment with cordycepin, when protected by an inhibitor of ADA, can prolong the survival of T. evansi-infected rats and provide curative efficacy. PMID- 21620641 TI - [Efficacy and patterns of ambulatory oxygen usage - experience of a university hospital]. AB - AIMS: To determine patterns of ambulatory oxygen (AO) use among patients with chronic obstructive pulmonary disease (COPD) and interstitial lung diseases, and analyze the effects of this therapy on daily activities and quality of life (QoL). PATIENTS AND METHODS: We included 37 consecutive adult patients on AO by liquid O(2) for more than three months prescribed by hospital pulmonologists. The acute response to O(2) was evaluated through the standardized 6-minutes walk test (6MWT) and the Borg dyspnea scale during the O(2) pre-intervention trial. Time spent away from home, compliance, side effects and QoL (SF-36 v1 questionnaire) were evaluated by a telephone interview during the follow-up period. Time spent away from home and QoL comparisons after and before the intervention were assessed retrospectively. RESULTS: COPD was the most frequent diagnosis (54%), and 29 (78%) patients were already on long-term oxygen therapy. In relation to the acute response to O(2) evaluated through the 6MWT, there were significant improvements in the distance walked (p<0.001), in resting SatO(2) (p<0.001), in minimal SatO(2) (p<0.001), and in percentage of desaturation (p=0.002), independently of the diagnosis. No differences were observed in Borg dyspnea scale. AO was used for a mean of 4.1h/day. Patients spent fewer hours per day away from home after AO treatment (3.5h vs. 5.0h, p<0.025). Six patients (16%) were not compliant to the prescription, and 54% mentioned side effects. We verified low scores in almost all of the sub-domains of SF-36 QoL questionnaire, with a significant improvement noted only in role emotional (p=0.032). Improvement in health global state was described by 49% of patients. CONCLUSIONS: Acute improvement in 6MWT parameters was not predictive of enhancement of outdoor activities and QoL with AO. More detailed studies are needed to achieve evidence based AO benefits. PMID- 21620642 TI - Measuring the quality of dying and quality of care when dying in long-term care settings: a qualitative content analysis of available instruments. AB - CONTEXT: Long-term care (LTC) settings have become a significant site for end-of life care; consequently, instruments that assess the quality of dying and care may be useful in these settings. OBJECTIVES: To evaluate the content of available measurement instruments to assess the quality of dying and care when dying. METHODS: Qualitative content analysis to categorize items as structure of care, process of care, satisfaction with health care (the first three representing quality of care and its evaluation), quality of dying, or patient factors. RESULTS: Instruments that measure mostly quality of care and its evaluation are the Family Perception of Physician-Family Caregiver Communication, End-of-Life in Dementia (EOLD) Satisfaction With Care, Family Perception of Care Scale, Toolkit of Instruments to Measure End-of-Life Care after-death bereaved family member interview (nursing home version), and the Family Assessment of Treatment at the End-of-Life Short version. Instruments measuring quality of dying are the EOLD Comfort Assessment in Dying, EOLD-Symptom Management, Mini-Suffering State Examination, and Palliative Care Outcome Scale. The Quality of Dying in Long-Term Care measures care and dying. The Minimum Data Set-Palliative Care measures mostly dying and patient factors. The instruments differ in dementia specificity, time of administration, and respondent. CONCLUSION: Instruments that assess quality when dying differ in several ways and most do not measure a single construct, which is relevant to guiding and evaluating care. Comparing psychometric properties and usefulness of instruments that measure similar constructs is the next step in determining which are best suited for use in LTC. PMID- 21620643 TI - The use of palliative care services associated with better dying circumstances. Results from an epidemiological population-based study in the brussels metropolitan region. AB - CONTEXT: There is some consensus that a "good death" is one which occurs at home, in the presence of loved ones, and is free of distress. Involvement of palliative care services is assumed to improve these circumstances. OBJECTIVES: This population-based study describes characteristics of the end of life of patients dying in Brussels and examines their associations with the involvement of palliative care services. METHODS: In 2007, an anonymous large-scale mortality follow-back survey was conducted in Brussels by mailing questionnaires regarding end-of-life care and characteristics to the attending physicians of a representative sample of 1961 deaths. RESULTS: Response rate was 41% (n=701). Of all deaths, 59% were nonsudden. Of these, 12% took place at home. For 27%, the attending physician was informed about the patient's preferred place of death. When the preference to die at home was known, 66% died at home. At the moment of death, in 47% of nonsudden deaths, loved ones were present. In a quarter of deaths, palliative care services were involved. Involvement with palliative care services was more likely for cancer patients than noncancer patients and was associated with the attending physicians more often being informed about the preferred place of death, with patients more often dying at home, with loved ones more often being present, and with a better feeling of well-being on the last day. CONCLUSION: Overall, circumstances at the very end of life are suboptimal in Brussels. This study provides a case for stronger involvement of palliative care as a focal point for policies to improve end-of-life circumstances in a metropolitan area such as Brussels. PMID- 21620644 TI - Nonnutritive sucking and oral sucrose relieve neonatal pain during intramuscular injection of hepatitis vaccine. AB - CONTEXT: Newborns are subject to pain during routine invasive procedures. Pain caused by immunization injections is preventable, but remains untreated in neonates. OBJECTIVES: The purpose of the study was to compare the effectiveness of three nonpharmacological pain relief strategies on newborns' pain, physiological parameters, and cry duration before, during, and after hepatitis B intramuscular (IM) injection. METHODS: In this prospective, randomized clinical trial, we enrolled 165 newborns (gestational age, >=36 weeks). The infants received IM injections and were randomized to three treatment groups: nonnutritive sucking (NNS), 20% oral sucrose, or routine care. Pain was measured by the Neonatal Facial Coding System, physiological signals by electrocardiogram monitors, and cry duration using a stopwatch. RESULTS: Pain was significantly lower among infants in the NNS (B=-11.27, P<0.001) and sucrose (B=-11.75, P<0.001) groups than that in controls after adjusting for time effects, infant sleep/wake state, number of prior painful experiences, and baseline pain scores. Infants in the NNS and sucrose groups also had significantly lower mean heart and respiratory rates than the controls. Cry duration of infants receiving sucrose was significantly shorter than those in the NNS (Z=-3.36, P<0.001) and control groups (Z=-7.80, P<0.001). CONCLUSION: NNS and oral sucrose can provide analgesic effects and need to be given before painful procedures as brief as a one-minute IM injection. Sucrose orally administered two minutes before injection more effectively reduced newborns' pain during injection than NNS. Both nonpharmacological methods more effectively relieved newborns' pain, stabilized physiological parameters, and shortened cry duration during IM hepatitis injection than routine care. PMID- 21620645 TI - Implementation of the victoria bowel performance scale. AB - CONTEXT: There is a lack of evidence to guide constipation management in patients receiving palliative care. Data collection requires the systematic use of validated assessment tools. OBJECTIVES: The objective of this study was to assess the usefulness of the Victoria Bowel Performance Scale (BPS) as an audit tool. METHODS: Charts were reviewed before and after the implementation of a program to monitor constipation through repeated use of the Victoria Bowel Scale. The program was initiated at three oncology pain and symptom management clinics, four palliative care units, and four residential hospices. An additional "control" palliative care unit introduced new nursing assessment tools without the new scale. RESULTS: The Victoria BPS was recorded at 86% of 192 postimplementation outpatient clinic visits and was easy to use in this setting. Documentation of bowel performance at comparable visits improved from 44% to 66% (P<0.001), and the frequency of changes to laxatives increased from 14% to 39% of visits (P<0.001). The scale was completed on 21%-55% of inpatient days, and variations in the proportion of recordings being rated as satisfactory between -1 and +1 (possible range from -4 to +4) revealed important deficiencies in bowel care, which led to change in management. CONCLUSION: The Victoria BPS was found to be an acceptable and a useful bowel function assessment tool, uniquely incorporating the patient's usual bowel function. Modifications to the scale have been made to improve clarity and allow for the expected drop in bowel activity seen in end-of life care. Considerable educational effort and appropriate organization of the charts are required for optimal implementation. The proportion of revised BPS scores ranging from -1 to +1 is proposed as an indicator of satisfactory bowel management for clinical, audit, and research purposes. PMID- 21620646 TI - Navigating tensions: integrating palliative care consultation services into an academic medical center setting. AB - CONTEXT: Despite rapid proliferation of hospital-based palliative care consultation services (PCCSs) across the country, there is little description of the dynamic processes that the PCCS and the non-PCCS hospital cultures experience during the institutionalization of a successful PCCS. OBJECTIVES: To describe the institutionalization of a new PCCS in a quaternary care academic medical center (AMC) and highlight two themes, cost and quality, that pervaded the dynamics involved from the inception to the successful integration of the service. METHODS: Ethnography using longitudinal field observations, in-depth interviews, and the collection of artifacts. The study was performed in a 750-bed quaternary care AMC in the northeastern region of the U.S. Participants were a purposefully selected sample (n=79) of 1) senior-level institutional administrators, including clinical leaders in nursing, medicine, and social work, 2) clinicians who used the PCCS, either commonly or rarely, and 3) members of the PCCS core and extended teams. RESULTS: Key infrastructure components that contributed to the successful integration of the PCCS included top level interprofessional administrative buy in to the quality and cost arguments for PCCS, PCCS leadership selection, robust data collection strategies emphasizing quality outcome data, the adoption of the "physician referral only" rule, and incremental and sustainable growth. The PCCS service grew an average of 23% per year from 2003 to 2009. CONCLUSION: An in depth understanding of the dynamic interaction of the infrastructures and processes of a successful institutionalization, in their unique complexity, may help other PCCSs identify and negotiate attributes of their own circumstances that will increase their chances for successful and sustainable implementation. PMID- 21620647 TI - What palliative care-related problems do patients experience at HIV diagnosis? A systematic review of the evidence. AB - CONTEXT: Palliative care is an essential element of HIV care throughout the disease trajectory, but there is a lack of information to guide clinical care at HIV diagnosis. OBJECTIVES: This systematic review aimed to identify and appraise the evidence of palliative care-related problems at HIV diagnosis. METHODS: The search strategy combined the term "HIV" with seven key words derived from the World Health Organization definition of multidimensional palliative care, in a systematic search of four databases. Abstracts and papers were screened to identify those recording problems within six months of HIV diagnosis in adults. Sample descriptions, aims, methods, and prevalence findings were extracted from these papers into common tables. RESULTS: Of 5443 titles retrieved, 65 met the inclusion criteria and 34 were retained. Papers included 27 original studies and seven secondary analyses of patient's records, with great heterogeneity in design, sample definition, and outcome measures. Physical and psychological symptoms were highly prevalent (pain 11%-76%, weight loss 8%-89%, fever 32%-89%, diarrhea 6%-54%, anxiety 36%-95%, and depression 18%-47%). At HIV diagnosis, well being was impaired, suicidal thoughts were frequent, and peace and calmness were reduced. Participants lacked emotional support and feared the reaction of their families. Practical problems included hunger, homelessness, reduced ability to work, and need for childcare. Studies had methodological failings such as the use of unvalidated tools and lack of clarity reporting results. CONCLUSION: People who have recently been diagnosed with HIV have multidimensional palliative care related problems. HIV care and support services need to assess and manage problems using integrated palliative care, with referral for complex problems. Patient centeredness must be a principle of HIV clinical research. PMID- 21620648 TI - Prevalence and predictors of sleep difficulty in a national cohort of women with primary breast cancer three to four months postsurgery. AB - CONTEXT: Mounting evidence suggests that many cancer patients suffer from sleep difficulty, but there is conflicting evidence regarding the prevalence and predictors of this adverse symptom. OBJECTIVES: The present study investigated the prevalence and predictors of clinically significant sleep difficulty in women with primary breast cancer. METHODS: Danish women (n=3343) with primary breast cancer completed the Pittsburgh Sleep Quality Index (PSQI) along with measures of depression, anxiety, physical activity/functioning, and health behaviors three to four months postsurgery. Data on disease status, treatment, and comorbidity were obtained from the Danish Cancer Cooperative Group and surgical departments, and information on sociodemographic factors and psychiatric history was obtained from Danish national longitudinal registries. RESULTS: More than half (57.9%) of the women reported clinically significant sleep difficulty (PSQI >5). Multiple logistic regression identified seven significant predictors of sleep difficulty in the full sample. In order of strength, these were the following: more depressive symptoms, poorer physical functioning, older age, higher levels of trait anxiety, consuming more cigarettes, having undergone lumpectomy, and lower levels of physical activity. Subgroup analysis found that more depressive symptoms and poorer physical functioning were the only two predictors that were significant in both pre- and postmenopausal women. CONCLUSION: These findings indicate that a high proportion of women with breast cancer experience sleep difficulty. Depression and poorer physical functioning appear to be robust predictors of sleep difficulty, whereas other predictors may depend on sample characteristics, including menopausal status. PMID- 21620649 TI - Challenging neck anatomy is associated with need for intraoperative endovascular adjuncts during endovascular aortic aneurysm repair (EVAR). AB - BACKGROUND: The purpose of this study was to determine which proximal seal zone characteristics were predictive of early and late type Ia endoleak development after endovascular aortic aneurysm repair (EVAR) for infrarenal abdominal aortic aneurysmal disease. METHODS: We evaluated 146 patients who underwent EVAR between January 2006 and March 2007. In the cohort, high-resolution computed tomography images of 100 (68.5%) patients were available, which showed detailed measurement of proximal neck parameters, including diameter, length, calcification, thrombus, suprarenal and infrarenal angles, and reverse taper morphology. Postprocessing of digital data sets was performed to obtain centerline-of-flow measurements. Relevant medical records and follow-up computed tomography scans were reviewed. RESULTS: Mean age of the patients was 72.7 years, with 78% being male. Of these patients, 66% did not satisfy the instructions for use for the Zenith EVAR device, and 50% did not satisfy the instructions for use for the AneuRx device. Nine patients had intraoperative type Ia endoleaks. A 100% assisted primary technical success rate was achieved with the adjunctive use of angioplasty (n = 4), uncovered stent (n = 3), and extension cuff (n = 2) placement. There was a significant association between type Ia endoleak development and magnitude of the infrarenal angle (p < 0.01); however, other parameters were not significant. At follow-up (mean, 587 days), no patient had a type Ia endoleak, and there were no aneurysm-related deaths. CONCLUSIONS: Our data indicate that infrarenal angle is related to intraoperative type Ia endoleak occurrence, but other factors often thought to be indicative of adverse neck anatomy are not significant predictors. Moreover, all type Ia endoleaks in this cohort were successfully eliminated intraoperatively, and durability was confirmed on postoperative surveillance. These data demonstrate that challenging neck anatomy is associated with the need for intraoperative endovascular adjuncts, and that effective and durable aneurysm exclusion should still be expected. PMID- 21620650 TI - Protein S-100B as biochemical marker of brain ischemic damage after treatment of carotid stenosis. AB - BACKGROUND: S-100 protein is a family of low molecular weight proteins found in vertebrates characterized by two calcium binding sites of the helix-loop-helix ("EF-hand type") conformation. There are at least 21 different types of S-100 proteins. The name is derived from the fact that the protein is 100% soluble in ammonium sulfate at neutral pH. Protein S-100B was investigated as a marker of brain ischemic damage after treatment of carotid stenoses. METHODS: Between December 1, 2009 and December 1, 2010, S-100B protein was monitored in 76 patients after carotid artery stenting (CAS) and in 24 patients after carotid endarterectomy (CEA). In each patient, multiple samples were taken: before the procedure (basal sample), immediately after CAS or CEA, 60 minutes after CAS or CEA, and daily during the hospital stay. Evaluation of S-100B was carried out by blind assessment. Patients underwent pre- and postoperative diffusion-weighted magnetic resonance imaging or computed tomographic scan. RESULTS: An S-100B coefficient of variation higher than the established cut-off was detected in 16 patients: three affected by postoperative stroke, two patients with minor stroke, and one patient with fatal stroke; 12 patients presented with uneventful neurological outcome and positive brain imaging; and there was one false positive case. No false negative cases occurred. The postoperative protein S-100B level lowered to basal level in 15 patients: within 24 hours in the 12 patients with the uneventful outcome (and positive brain imaging) and in the false positive case; and after 120 and 144 hours, respectively, in the two patients with minor stroke. In the patient with fatal stroke, protein S-100B never returned to the preoperative level. CONCLUSIONS: In patients with an increased S-100B coefficient of variation, the diffusion-weighted magnetic resonance imaging was positive for ischemic brain lesions, except for one patient who was reported as a false positive case. The postoperative S-100B protein level decreased within 24 hours in the uneventful neurological cases and in the false positive case, whereas long lasting postoperative increased values of the S-100B protein were observed in patients with poor neurological outcomes. PMID- 21620651 TI - Late complication of thoracic endovascular stent-grafting. AB - Thoracic endovascular aortic repair (TEVAR) has become increasingly popular for the treatment of descending thoracic aortic aneurysms. Despite their proven efficacy and safety, several complications can occur with TEVAR. We report a case of a 76-year-old woman with a late-term complication after TEVAR related to recurrent thyroid cancer and carotid-subclavian revascularization. PMID- 21620652 TI - Cardiovascular prognosis at 1-year of patients with acute coronary syndrome is related to abdominal aortic aneurysm despite small size of the aneurysm. AB - BACKGROUND: Global cardiovascular (CV) risk associated with abdominal aortic aneurysms (AAAs) has been poorly documented. The aim of this study was to evaluate whether the presence of AAA in patients hospitalized for acute coronary syndrome with coronary stenosis (>=50%) was associated with an increased CV risk at 1-year follow-up. METHODS: Between February 1, 2008 and March 30, 2009, 304 patients admitted for acute coronary syndrome with significant (>=50% stenosis) coronary lesions underwent echocardiography to check for presence of AAA. Twenty AAAs were diagnosed, of average (+/-standard deviation) diameter 33 +/- 3.7 mm. Follow-up at 1 year was available for 288 patients (95%). Variables recorded at 1 year were death, cause of death, and occurrence of nonfatal CV events of cardiac or peripheral vascular origin. RESULTS: During follow-up, 65 patients (22.6%) experienced an event (all-cause death or nonfatal CV event), including 21 deaths (7.3%) and 44 nonfatal CV events (15.3%). The presence of AAA significantly increased the risk of any CV event (fatal or nonfatal) at 1 year (hazard ratio: 2.96, 95% CI: 1.49-5.89, p = 0.002) but did not influence overall mortality or CV mortality. CONCLUSION: Our results show that in patients with coronary artery disease already at high CV risk, the presence of AAA was associated with worse CV prognosis at 1 year, and incurred an increased risk of occurrence of any CV event (fatal and nonfatal). PMID- 21620653 TI - Endovascular repair of a ruptured abdominal aortic aneurysm in a patient with bilateral iliac dissections. AB - BACKGROUND: The presentation of a ruptured fusiform abdominal aortic aneurysm (AAA) co-occurring with bilateral iliac dissections is extremely rare. Endovascular repair of ruptured AAA is an accepted treatment modality for suitable patients; however, this approach may be complicated by the presence of iliac arterial pathology. METHODS AND RESULTS: We report the case of a 66-year old man who presented with a ruptured AAA. Preoperative imaging demonstrated bilateral iliac artery dissections in addition to the ruptured AAA. We describe the technical aspects of the combined endovascular and open surgical repair required to manage this complex presentation. CONCLUSION: The combined presentation of ruptured AAA and bilateral iliac dissections has not been previously reported. Successful repair can be achieved using a combined endovascular and open surgical approach. PMID- 21620654 TI - Prevention of pulmonary complications after aortic surgery: evaluation of prophylactic noninvasive perioperative ventilation. AB - BACKGROUND: In 40% of the cases, pulmonary complications are encountered after aortic surgery in patients suffering from chronic obstructive broncho-pneumopathy (COBP). The factors aggravating the occurrence of these complications are cumulated tobacco addiction of >=40 packets per year and surgery for aneurysm. The aim of our prospective study was to evaluate the effect of pre- and postoperative noninvasive ventilation (NIV) preparation on the respiratory function of patients presenting with high pulmonary risks. METHODS: Between September 2007 and May 2010, 30 patients were included in the present study. Inclusions criteria were male gender, patients suffering from COBP with tobacco addiction of >=40 packets per year, and aortic surgery for aneurysm with or without occlusive lesions. In all, 14 of the 15 patients had effectively performed the preparation before and after surgery according to a fixed protocol (NIV group). We compared the respiratory rate complications and the mean hospital length of stay in intensive care with the non-NIV control group (15 patients). RESULTS: Mean age, severity of COBP according to the classification of the French Society for Pneumology, surgery duration, and blood losses were comparable between the two groups. Conversely, pulmonary complications were significantly lower in the NIV group (0/14 [0%] vs. 5/15 [33%], p = 0.004). Hospital length of stay in intensive care was shorter in the NIV group (2.5 vs. 6.5 days, p < 0.001). CONCLUSION: Preliminary results are encouraging, in favor of pulmonary preparation before aortic surgery with NIV at home, and carried on postoperatively in hospital for patients with COBP. These results prompt us to propose a multicenter study to validate these first observations. PMID- 21620655 TI - Intraoperative evaluation of blood perfusion by laser-assisted indocyanine green angiography after ex vivo vascular reconstruction of intrahilar renal artery aneurysm. AB - The surgical reconstruction of intrahilar renal artery aneurysms (RAAs) is a difficult surgery because of complex anatomy. We present a case of right intrahilar RAA diagnosed in a 67-year-old man. We performed ex vivo reconstruction using an organ preservation solution to prevent postoperative renal failure. We assessed graft patency and blood perfusion was assessed by laser-assisted indocyanine green angiography using the SPY system after autotransplantation. Postoperative renal insufficiency was not observed. The results demonstrate that ex vivo reconstruction of intrahilar RAAs using an organ preservation solution, and graft patency and blood perfusion evaluation using the SPY system are effective methods for preserving renal function. PMID- 21620656 TI - Inflammatory markers and restenosis in peripheral percutaneous angioplasty with intravascular stenting: current concepts. AB - In this article, we review the current status of inflammation linked to percutaneous transluminal angioplasty with stent implantation, especially as it relates to restenosis and its clinical implications. Common to multiple vascular diseases, including atherosclerosis, interventional restenosis is a localized inflammatory reaction. Activated smooth muscle cells respond to local inflammation and migrate from the media into the lumen of the vessel, where they proliferate and synthesize cytokines which they respond to in an autocrine manner, sustaining the progression of the lesion. The deleterious effects of proinflammatory cytokines, particularly immunomodulatory interleukins, on vascular pathophysiology and development of these maladaptive processes have been the subject of intense study. Matrix metalloproteinases and tissue inhibitors of metalloproteinases are important in many physiologic and pathologic processes and their expression is related with the classic cardiovascular risk factors as well as with inflammation. They seem to play a central role in atherosclerosis and restenosis. The primary use of drug-eluting stents has become routine clinical practice for coronary artery disease, but the use in peripheral arteries remains to be further studied, like that demonstrated in sirolimus-coated Cordis trials. New studies to understand this complex process in peripheral arteries are warranted. PMID- 21620657 TI - Treatment of a recurrent false aneurysm of the femoral artery by stent--graft placement from the brachial artery. AB - An anastomotic false aneurysm is a well known complication after femoral artery surgery. Open surgical repair is the treatment of choice for anastomotic femoral aneurysms, but this can be challenging, unsuccessful, or even impossible. Endovascular repair is an alternative in these cases, but the delivery of a stent -graft in the femoral artery can be difficult. We report the case of a patient with a recurrent left femoral artery anastomotic false aneurysm, treated twice by open exclusion, and finally excluded successfully by a stent--graft that was inserted through the left brachial artery. PMID- 21620658 TI - Antegrade, covered, self-expanding stent as an iliac extension in a bifurcated endograft: a feasible technical maneuver for challenging aortoiliac aneurysmal anatomy. AB - BACKGROUND: Aneurysmal and occlusive aortoiliac disease can make the process of introducing large delivery catheters for endovascular repair challenging. We describe the case of a patient who could be treated by a bifurcated stent-graft despite having a unilateral external iliac occlusion. METHODS AND RESULTS: From a brachial access, a covered self-expanding stent was deployed antegradely through the distal gate of the stent-graft into the common iliac artery. This technical choice helped to overcome the problem of an external iliac occlusion, so as to maintain an antegrade flow into the internal iliac and avoid the need for an interfemoral bypass. CONCLUSION: Auto-expandable covered stent-graft with a thinner shaft can be used through a brachial access as an iliac extension of a bifurcated aortic endograft. However, a longer follow-up duration and more cases are necessary to warrant the safety and the durability of such an "off-label" endovascular material assemblage. PMID- 21620659 TI - Aneurysm of the aberrant right subclavian artery: surgical and hybrid repair of two cases in a single center. AB - The aberrant right subclavian artery (ARSA) aneurysm is rare; however, the risk of rupture and thromboembolism is high, with a postrupture mortality rate of 50%. In this report, we have described two cases of this anomaly. In the first case, a 62-year-old male patient presented with a symptomatic aneurysm of ARSA (maximum diameter of 4 cm) causing chest pain with dyspnea during moderate physical effort. Surgical treatment was performed with aneurysmal exclusion and direct anastomosis of the two heads of the subclavian artery. In the second case, a 72 year-old male patient presented with a symptomatic aneurysm (maximum diameter of 5.1 cm) of ARSA causing dysphagia and dysphonia. In this case, a one-stage hybrid treatment was performed: a bilateral carotid-subclavian bypass was associated with intentional occlusion of both subclavian arteries (by plug positioning) during thoracic endovascular stent-grafting procedures. In both cases, the peri/postoperative course was uneventful and the technical results in our series were excellent at long-term follow-up. Surgical treatment can be safely performed in patients with low operative risk or whenever endovascular technique is not suitable. The ARSA aneurysm, with appropriate anatomy, can be successfully treated by hybrid treatment (combined surgical and endovascular approach). We reckon that this minimally invasive technique helps avoid thoracotomy and could be the treatment of choice in high-risk patients. PMID- 21620660 TI - Manual high-dose regional intraoperative thrombolysis of lower extremity emboli after open repair of a ruptured abdominal aortic aneurysm. AB - We describe a case of high-dose regional intraoperative thrombolysis subsequent to mechanical thrombectomy for the treatment of postoperative distal extremity embolization of the right lower limb owing to open repair of a ruptured abdominal aortic aneurysm. Mechanical thrombectomy was performed from the popliteal artery but residual embolic occlusion of all three tibial arteries remained. The limb was elevated, exsanguinated, and a blood cuff was placed below the knee and inflated to suprasystolic pressure to isolate the limb from systemic circulation. An 18-gauge infusion catheter was introduced to the exposed dorsalis pedis artery. Subsequently, the exposed great saphenous vein was cannulated and drained. A total of 100 mg of recombinant tissue-type plasminogen activator diluted in 500 mL of saline was infused into the anterior tibial artery with a slow hand infusion for 30 minutes. The infusion was continuously collected through the great saphenous cannulation and a closed loop was confirmed by angiogram. The limb was flushed with heparin and saline solution. Infusion catheter was extracted and the great saphenous vein was ligated. Blood cuff was removed, arterial flow was re-established, and a postprocedural arteriogram confirmed successful revascularization. This method may be an alternative to microtibial embolectomy at the foot ankle level after severe lower limb embolization after acute open repair of a ruptured abdominal aortic aneurysm. PMID- 21620661 TI - Type B aortic dissection resulting in acute esophageal necrosis. AB - In the present article, we report a case in which acute esophageal necrosis (AEN) of the intrathoracic esophagus was caused by extensive thrombosis in the false lumen of an aortic dissection, thereby occluding the blood flow to the intercostal arteries and thus the esophagus. According to the previously published data, AEN after aortic dissection is very rare and usually fatal. Besides esophageal ischemia secondary to arterial occlusion, direct extrinsic compression of the arteriovenous network surrounding the esophagus, caused by the traumatic pathology of the aorta, by extensive extravasation may also cause AEN. AEN is most commonly confirmed by esophagoscopy, typically showing a black, diffusely necrotic, and ulcerated esophageal mucosa. PMID- 21620662 TI - Endovascular stent--graft repair of a symptomatic superior mesenteric artery aneurysm. AB - BACKGROUND: Aneurysms of the visceral branches of the abdominal aorta are uncommon and potential life-threatening entities. Although hepatic and splenic artery aneurysms have the highest prevalence among splanchnic aneurysms, superior mesenteric artery aneurysms are even more uncommon. For ruptured visceral arteries aneurysms, a mortality rate of between 20% and 100% has been reported. Besides rupture, the aneurysms can also erode into adjacent visceral organs, which results in severe hemorrhage. Emergency surgery of visceral artery aneurysms is related with significant mortality. In recent years, endovascular therapies have been successfully established in the elective setting. Therefore, we adopted the endovascular stent-graft insertion technique for dealing with an acute symptomatic aneurysm of the superior mesenteric artery. METHOD: We report the case of a 79-year-old male patient with a symptomatic aneurysm of the proximal superior mesenteric artery. Using an endovascular approach, the aneurysm was excluded with an endoluminal stent--graft. RESULT: The endoluminal stent- graft repair resulted in total exclusion of the aneurysm. Patency of the superior mesenteric artery continued and no ischemic complications occurred. Pain and other symptoms disappeared and the patient recovered entirely. CONCLUSIONS: Endovascular management of symptomatic superior mesenteric artery aneurysm is feasible and may display excellent results in selected cases. Therefore, endoluminal stent--graft repair should be considered as an alternative to traditional surgical treatment. However, in each patient, the individual anatomy of the mesenteric circulation and the location of the aneurysm play a major role in determining whether endovascular management is possible or surgical repair should be preferred. PMID- 21620663 TI - Thrombectomy and surgical reconstruction for extensive iliocaval thrombosis in a patient with agenesis of the retrohepatic vena cava and atresia of the left renal vein. AB - In 80% of the patients presenting with deep-venous thrombosis (DVT), a risk factor can be identified. An absent or hypoplastic infrarenal vena cava is a rare risk factor for DVT in young adults. In these cases, the prevalence of congenital anomalies of the inferior vena cava (IVC) is estimated at 0.5% of the general population, up to 5% in young people. The association with coagulopathy increases the risk of DVT. We report a case of a young man who presented with a massive caval and iliofemoral-popliteal thrombosis in presence of the agenesis of retrohepatic inferior vena cava and atresia of the left renal vein. Open thrombectomy and caval reconstruction with a polytetrafluoroethylene graft were performed. Surgical option with vein reconstruction was preferred to prevent new episodes of thrombosis and the risk of acute renal failure. PMID- 21620664 TI - Chronic rupture of abdominal aortic aneurysm. AB - Although the mortality rate after abdominal aortic aneurysm rupture approximates 90% despite the urgent management, a few cases of chronic rupture and delayed repair have been reported in the world literature; anatomic and hemodynamic reasons occasionally allow for the fortunate course of these patients. We report in this article the case of 76-year-old man with a ruptured abdominal aortic aneurysm who was transferred to our facility 4 weeks after his initial hospitalization in a district institution and who finally had a successful open repair. PMID- 21620665 TI - Infusion of recombinant human tissue plasminogen activator through the superior mesenteric artery in the treatment of acute mesenteric venous thrombosis. AB - Acute mesenteric venous thrombosis is an uncommon condition that is usually treated with systemic anticoagulation. Catheter-directed thrombolysis through the superior mesenteric artery may be a viable adjunct to treat this morbid condition. In the present article, we have described a case of superior mesenteric venous thrombosis treated with catheter-directed infusion of tissue plasminogen activator through the superior mesenteric artery. PMID- 21620666 TI - Surgery for acute lower limb ischemia in the elderly population: results of a comparative study. AB - BACKGROUND: To evaluate the outcome of surgical treatment of acute lower limb ischemia in elderly patients. The primary endpoints were early and midterm rates of survival and limb salvage. METHODS: A retrospective analysis involving 76 consecutive patients observed for acute lower limb ischemia between March 2005 and December 2008 who underwent revascularization was conducted. A total of 44 patients (group A) of age >80 years (average age: 86.9 +/- 4.5 years; 13 men and 31 women) had a site of obstruction at the level of abdominal aorta in one case (2.3%), common-external iliac artery/common femoral artery in 15 (34.1%), superficial femoral artery/popliteal artery in 26 (59.1%), and infrapopliteal arteries in two (4.5%), and embolectomy, thrombectomy, bypass, and angioplasty was performed in 24 (54.5%), 11 (25.0%), eight (18.2%), and one (2.3%) case(s), respectively. A total of 32 patients (group B) of age <80 years (average age: 67.2 +/- 12.2 years; 19 men and 13 women) with an analogous site of obstruction at the level of abdominal aorta in one case (3.1%), common/external iliac artery/common femoral artery in 12 (37.5%), superficial femoral artery/popliteal artery in three (9.4%), and infrapopliteal arteries in 16 (50.0%) underwent the same aforementioned procedures in 10 (31.3%), 12 (37.5%), nine (28.1%), and one (3.1%) case(s), respectively. RESULTS: At 30 days after surgery, survival and limb salvage were both 93.2% in group A versus 96.9% and 87.5%, respectively, in group B, whereas they were 93.2% and 65.8%, respectively, in group A versus 85.7% and 96.9%, respectively, in group B (p = 0.22 and p = 0.19, respectively) at the midterm control. The univariate and multivariate analyses showed that category 2b of acute ischemia (immediately-threatening) has a negative statistically significant influence on the survival rate (p = 0.01). CONCLUSIONS: Limb salvage rates after surgical intervention for acute lower limb ischemia in the elderly population are comparable with those of the younger population, whereas survival rates are lower, but without statistical significance. The only factor that negatively influences the survival rate is the 2b clinical category of acute ischemia, regardless of the age of the patient. PMID- 21620667 TI - Popliteal pseudoaneurysm caused by stent fracture. AB - Stent fracture with pseudoaneurysm formation in the femoropopliteal artery has uncommonly been reported. We present the case of a 72-year-old man with a fracture of self-expanding nitinol stent and a pseudoaneurysm formation in the suprageniculate popliteal artery. The popliteal artery was successfully reconstructed with a small saphenous vein graft interposition. PMID- 21620668 TI - Successful embolization of a suprascapular artery aneurysm. AB - A 45-year-old woman was referred to our service because 9 months earlier she had developed a pulsating mass on the right supraclavicular fossa and torticollis. Ultrasounds and computed tomographic arteriography showed the presence of a subclavian collateral artery aneurysm with a diameter of 21 mm. On selective arteriography, an aneurysm of a suprascapular artery arising directly from the right subclavian artery was reported. The presence of thoracic outlet syndrome was excluded. The aneurysm was successfully treated with ethylene-vinyl alcohol polymer, a liquid embolic agent. The patient was discharged on postoperative day 1 in good general condition. After 12 months, control ultrasounds confirmed the complete thrombosis of the aneurysm sac. PMID- 21620669 TI - Heparin-induced thrombocytopenia type II because of heparin-coated polytetrafluoroethylene graft used to bypass. AB - Heparin-induced thrombocytopenia type II is an immune-mediated syndrome that may arise in a time-dependent manner after heparin therapy. Thrombocytopenia and thrombosis in patients exposed to heparin are suggestive of this syndrome. PMID- 21620670 TI - Single-center experience with open surgical treatment of 36 infected aneurysms of the thoracic, thoracoabdominal, and abdominal aorta. AB - BACKGROUND: To describe a single-center experience with open surgical treatment of infected aortic aneurysms. We analyzed risk factors for 90-day mortality. METHODS: Between 1983 and 2008, 4,410 patients underwent open surgery for thoracic, thoracoabdominal, or abdominal aneurysm at our institution. Primary infection of the aneurysm was suspected because of clinical signs of infection in combination with typical radiological and morphological aspects in 66 patients (1.5%). In all, 36 patients displayed 10 different kinds of organisms in cultures of blood and/or intraoperative specimens and were further analyzed. RESULTS: The group consisted of 23 men and 13 women, with a mean age of 66.8 +/- 8 (50-84) years. Location of the aneurysm was thoracic in five patients (14%), thoracoabdominal in 13 patients (36%), and abdominal in 18 patients (50%). Eleven patients (28%) were treated before and 25 (72%) after 1995. We found free rupture in three cases; contained rupture into surrounding tissue in 23 cases (64%); penetration into lung, bronchus, esophagus, or inferior vena cava in five cases; and an intact aneurysm in another five cases. Kinds of surgery were as follows: extra-anatomic revascularization in four patients (11%), Dacron patch plasty in four patients (11%), in situ revascularization in 24 patients (66%), and four patients died during surgery before reconstruction (11%). In all, 13 patients died during hospital stay (36%). In 25 patients treated after 1995, 90-day mortality was 24% and was significantly better (p < 0.05) than the rate of 64% in 11 patients treated before 1995. Outcome depended on status of rupture: all patients with free rupture, three of five patients (60%) with rupture into an organ, seven of 23 patients (30%) with contained rupture into the surrounding tissue, but no patient with intact aneurysm died. Age, gender, bacterium, location of the aneurysm, and method of surgical treatment did not influence 90 day mortality. During follow-up, 18 patients died after a mean of 56 +/- 49 months. One patient died because of an infected aortic prosthesis. CONCLUSIONS: Outcome of patients with infected aortic aneurysms has improved during the last 15 years and depends on the status of rupture at time of surgery. Therefore, only early diagnosis and early treatment can further improve the prognosis. PMID- 21620671 TI - Is open repair still the gold standard in visceral artery aneurysm management? AB - BACKGROUND: Visceral artery aneurysms and pseudoaneurysms represent a rare disease with high mortality. The aim of this study was to report a single center experience of open repair (OR) and endovascular treatment (ET) of 94 patients, and to analyze short- and midterm results. METHODS: Between 1988 and 2010, 94 patients, 43 men and 51 women, mean age of 57.6 years (range, 23-87 years), were referred to our Institute with a diagnosis of visceral artery aneurysm or pseudoaneurysm. Arteries involved were splenic artery in 44 cases, hepatic artery in 17, renal artery in 18, superior mesenteric artery in six, celiac trunk in three, gastroduodenal in two, and pancreaticoduodenal in four. An abdominal aortic aneurysm coexisted in three (3%) cases, whereas in six (6%) cases, there were multiple visceral aneurysms. ET was indicated based on the anatomical location of the aneurysm or for patients at high risk for surgery. RESULTS: A total of 74 patients underwent OR, whereas ET was performed in 20 patients. Technical success was achieved in all cases treated by open surgery. Splenectomy was performed in 11 cases, and in six, splenic autotransplantation was performed. At 6 months of follow-up, a Tc99m-labeled red cell scintigraphy showed that autotransplants were viable in four patients (67%). No cases of pancreatitis or splenic infarction were observed. Among renal artery aneurysms, nephrectomy was necessary in one case of renal infarction for massive thrombosis of the ex-vivo reconstructed renal artery. Four surgical conversions were recorded (one thrombosis of the hepatic artery, one massive hemorrhage after embolization of superior mesenteric artery aneurysm, and two cases of sac enlargement after 24 and 48 months). An endoleak was present in a patient treated for a splenic artery aneurysm, but it resolved spontaneously after 6 months. No complications were observed in patients undergoing surgical conversion after ET. Perioperative mortality in the surgical group was 1.3% (1/74). There was no perioperative mortality in the endovascular group. No statistically significant difference was found between groups in terms of perioperative mortality (p = 1.00). Perioperative morbidity was 9.4% (7/74) in the surgical group, and 10% (2/20) in the endovascular group (p = 1.00). Follow-up was available for 16 patients in the endovascular group (80%) and 63 in the surgical group (85%), with a mean duration of 42 months (1-192 months). In the whole cohort, actuarial survival at 10 years was 68%. The Kaplan-Meier estimates of survival at 1 and 5 years were 100% and 85%, respectively, for OR, and 100% and 40%, respectively, for ET, with no significant difference between the two groups CONCLUSION: ET is safe and feasible in selected patients, but incomplete exclusion may be observed, requiring late surgical conversion in a significant number of patients. Long-term results (high survival, low complication rate) confirm the durability of the surgical approach that in our experience remains the gold standard with satisfactory results, especially for aneurysms involving the visceral hilum. PMID- 21620672 TI - Dose-dependent effect of rosuvastatin in the regulation of metalloproteinase expression. AB - BACKGROUND: The importance of rosuvastatin at therapeutic dosage in regulating the release, activity, protein level, and expression of matrix metalloproteinases (MMP)-2 and MMP-9 was investigated. METHODS: Human umbilical artery smooth muscle cells were stimulated, in vitro, in a serum-free medium with rosuvastatin at various concentrations (2, 4, 7, and 10 ng/mL, which correspond to the maximal plasma concentration observed in healthy men after a daily oral intake of 5, 10, 20, and 40 mg, respectively). The release of MMP-2 and MMP-9 in the conditioned medium was assessed by enzyme-linked immunosorbent assay and confirmed by Western blot, the activity and expression were determined by zymography and polymerase chain reaction, respectively. RESULTS: Human umbilical artery smooth muscle cells stimulated with rosuvastatin at 7 and 10 ng/mL had a significant lower release, activity, protein level, and expression of MMP-2 and MMP-9, when compared with those stimulated at 2 and 4 ng/mL (MMP-2 =p < 0.0001 and p < 0.0001, respectively; MMP-9 =p < 0.0001 and p < 0.0001, respectively). CONCLUSION: The effects of rosuvastatin in reducing MMP-2 and MMP-9, which might stabilize the atherosclerotic plaques, are dose-dependent. PMID- 21620673 TI - Abdominal aortic aneurysm repair complicated by infection with Clostridium septicum. AB - Aortic stent-graft infection after endovascular abdominal aortic aneurysm (AAA) repair is an uncommon, but very serious complication with potentially devastating consequences.(1) Traditional open techniques of repair of AAA demonstrate an infection rate of 0.5-3%. The exact rate of infection with endovascular repair is unknown, but literature review demonstrates an overall incidence of 0.43-1.17% retrospectively.(2,3) Etiology of endovascular graft infections typically results from flora derived from the skin or gastrointestinal tract.(4)Clostridium septicum is a naturally occurring anaerobic bacterium native to the gastrointestinal tract. It is typically associated with spontaneous nontraumatic gas gangrene owing to bacteremia from the gastrointestinal tract with an incidence rate of 0.07%.(5) To our knowledge, this is the first reported case of endovascular AAA graft infection owing to Clostridium septicum species. PMID- 21620674 TI - Thoracic endovascular aneurysm repair for thoracic aneurysms: what we know, what to expect. AB - Descending thoracic aneurysms are less common and less likely to rupture than abdominal aortic aneurysms. However, when left untreated they are lethal, and repairs are recommended only if the size of the aneurysm is 6 cm, or smaller if symptomatic. Open thoracic aneurysm repair was the standard therapy the last time this topic was reviewed in Annals of Vascular Surgery. Currently, thoracic aneurysms are usually treated with endovascular means when anatomically appropriate. This review demonstrates the benefits of endografting (namely decreased perioperative mortality and morbidity) and also the shortcomings (no late mortality benefit) as currently published in the literature, as well as summarizing available endografts and specific considerations for high-risk patient populations. PMID- 21620675 TI - Traumatic pseudoaneurysm of the visceral aortic segment managed using both open surgery and endovascular therapy. AB - Pseudoaneurysm (PSA) formation is not a rare entity in the thoracic aorta, but is much rarer in the visceral aortic segment. This vascular lesion is most commonly because of penetrating trauma. We present the case of a patient with PSA of the visceral aortic segment that was treated by both surgical and endovascular methods and review the previously published data on the presentation, diagnosis, and management of visceral aortic PSAs. PMID- 21620676 TI - Active cardiac model and its application on structure detection from early fetal ultrasound sequences. AB - The structure of an early fetal heart provides vital information for the diagnosis of fetus defects. However, early fetal hearts are difficult to detect due to their relatively small size and the low signal-to-noise ratio of ultrasound images. In this paper, a novel method is proposed for automatic detection of early fetal cardiac structure from ultrasound images. The proposed method consists of two major parts which are the preprocessing phase and the active cardiac model: (1) The preprocessing phase consists of two sub-steps. (a) The region of interest is first automatically selected based on an accumulated motion image, which is able to represent the motion information of the fetal heart more accurately. (b) Then by combining Rayleigh-trimmed filter and anisotropic diffusion in 3-dimensional space, a despeckling method is developed to suppress the speckle noise and emphasize the motion information for subsequent cardiac structure detection. (2) The active cardiac model is proposed for the detection of fetal heart structure, which is a key contribution of this paper. It takes into account both the structure and motion information of fetal hearts simultaneously. Both learning and inference of the active cardiac model are described in the paper. Experiments on seven ultrasound sequences demonstrate the effectiveness of the proposed method. PMID- 21620677 TI - Sliding reconstruction of the condyle using posterior border of mandibular ramus in patients with temporomandibular joint ankylosis. AB - The traditional approach for ankylosis is gap arthroplasty or interpositional arthroplasty followed by reconstruction of the condyle using, for example, costochondral grafts. As these are non-pedicled grafts, there is eventual resorption with subsequent decrease in height of the ramus, facial asymmetry and deviated mouth opening. The authors have applied the method of total and partial sliding vertical osteotomy on the posterior border of the mandibular ramus for reconstruction of the mandible condyle as a pedicled graft for the correction of temporomandibular joint (TMJ) ankylosis. From 2004 to 2008, 18 patients who were diagnosed with TMJ ankylosis underwent operations for resection of the ankylosed condyle. Two methods were performed depending on the level of osteotomy on the posterior part of the mandibular ramus. All patients were followed-up for an average of 36 months (range 24-48 months). All patients showed apparent improved joint function with no cases of re-ankylosis. The results showed that sliding vertical osteotomy on the posterior border of the mandibular ramus seems to be an alternative and promising method for condylar reconstruction in patients with TMJ bony ankylosis. PMID- 21620678 TI - Change in cardio-ankle vascular index by long-term continuous positive airway pressure therapy for obstructive sleep apnea. AB - BACKGROUND: It has been reported that patients with obstructive sleep apnea (OSA) have an elevated arterial stiffness, and alleviation of OSA by continuous positive airway pressure (CPAP) might attenuate this. Recently, the cardio-ankle vascular index (CAVI) has been reported to be a highly reproducible arterial stiffness parameter in OSA patients. However, the change in CAVI that occurs following long-term CPAP treatment for OSA remains unclear. METHODS: Patients with moderate-to-severe OSA were enrolled. Changes in CAVI at 1 and 12 months after CPAP initiation (DeltaCAVI(1) and DeltaCAVI(12), respectively) were assessed. Factors associated with DeltaCAVI(1) and DeltaCAVI(12) were determined by multivariable regression analyses. RESULTS: Thirty subjects were assessed. CAVI was significantly reduced at 1 month compared with the baseline from 7.80 +/ 1.19 to 7.56 +/- 1.08 (p = 0.013). A non-significant reduction was observed at 12 months (7.72 +/- 1.18, p = 0.365 versus baseline) and CAVI had actually increased compared with that measured at 1 month. In multivariable analyses, DeltaCAVI(1) was inversely correlated with CPAP usage (coefficient: -0.500, p = 0.006) and was directly correlated with the change in the ratio of low frequency to high frequency in heart rate variability (coefficient: 0.607, p < 0.001), whereas DeltaCAVI(12) was related to the use of angiotensin-converting enzyme inhibitors (ACE-I) or angiotensin-II-receptor blockers (ARB; coefficient: 0.464, p = 0.013), was directly correlated with the change in hemoglobin A1c levels (coefficient: 0.644, p < 0.001), and was inversely correlated with the change in CPAP usage (coefficient: -0.380, p = 0.046). CONCLUSIONS: CAVI was significantly reduced by short-term CPAP and then slightly increased from 1 to 12 months, which was probably due to natural progression associated with the aging process. However, long-term CPAP treatment had the beneficial effect of maintaining CAVI below baseline levels when associated with the use of ACE-I/ARB, the control of blood glucose and the CPAP compliance. PMID- 21620679 TI - Long-term clinical outcomes after deferral of percutaneous coronary intervention of intermediate coronary stenoses based on coronary pressure-derived fractional flow reserve. AB - Coronary pressure-derived fractional flow reserve (FFR) has been used to evaluate functional severity of coronary artery stenoses. The cut-off point of 0.75 was considered to be the indication for percutaneous coronary intervention (PCI). In this study, we examined the prognosis of patients in whom PCI was deferred because the lesion was not significant by FFR (>=0.75). We measured FFR of 44 patients (50 lesions with angiographically intermediate stenoses by pressure wire between 2002 and 2009. Out of 44 patients (50 lesions), functionally non significant stenoses with FFR>=0.75 were 29 patients (33 lesions) and PCI was deferred. In the remaining 15 patients (17 lesions), FFR was <0.75 and PCI was performed. Patients were followed up for an average period of 53 months with endpoints of major adverse cardiac events (MACE; cardiac death, acute coronary syndrome, PCI, and coronary artery bypass grafting). The rate of MACE was 2/29 (6.9%) in patients with FFR>=0.75 and 2/15 (13.3%) in those with FFR<0.75, and it was not statistically different between the two groups. Since long-term clinical outcomes after deferral of PCI of intermediate coronary stenoses based on FFR were excellent (annual event rate 1.6%/year), FFR is a useful index to judge the indication of PCI and risk-stratify patients for MACE. PMID- 21620680 TI - Modeling epileptogenesis and temporal lobe epilepsy in a non-human primate. AB - Here we describe a new non-human primate model of temporal lobe epilepsy (TLE) to better investigate the cause/effect relationships of human TLE. Status epilepticus (SE) was induced in adult marmosets by pilocarpine injection (250mg/kg; i.p.). The animals were divided in 2 groups: acute (8h post-SE) and chronic (3 and 5 months post-SE). To manage the severity of SE, animals received diazepam 5min after the SE onset (acute group: 2.5 or 1.25mg/kg; i.p.; chronic group/; 1.25mg/kg; i.p). All animals were monitored by video and electrocorticography to assess SE and subsequent spontaneous recurrent seizures (SRS). To evaluate brain injury produced by SE or SRS we used argyrophil III, Nissl and neo-Timm staining techniques. Magnetic resonance image was also performed in the chronic group. We observed that pilocarpine was able to induce SE followed by SRS after a variable period of time. Prolonged SE episodes were associated with brain damage, mostly confined to the hippocampus and limbic structures. Similar to human TLE, anatomical disruption of dentate gyrus was observed after SRS. Our data suggest that pilocarpine marmoset model of epilepsy has great resemblance to human TLE, and could provide new tools to further evaluate the subtle changes associated with human epilepsy. PMID- 21620681 TI - Social cognition and neurocognition as predictors of conversion to psychosis in individuals at ultra-high risk. AB - BACKGROUND: While deficits in cognitive functions are frequently reported in psychotic disorders, further longitudinal research is needed to confirm the specific risk factors for the development of psychosis. We examined longitudinally the social-cognitive and neurocognitive function of individuals at ultra-high risk for schizophrenia who developed psychosis later as predictive markers. METHOD: The investigators studied 49 subjects at ultra-high risk (UHR) for psychosis and 45 healthy controls. The UHR subjects were followed for up 5.2 years (mean: 2.8 years) and 13 of these subjects developed psychosis. Theory of mind (ToM) tasks and neuropsychological tests were administered at baseline. Analyses compared the UHR patients who later developed psychosis, those who did not develop, and healthy controls. To examine the cognitive variables to predict transition to psychosis, Cox regression analyses were conducted. RESULTS: At baseline, we found significant differences among the three groups in social cognition according to the False Belief and cartoon tasks and in neurocognition according to tasks measuring executive function, working memory, verbal memory, and visual memory. Our study showed that a model combining working memory, visual memory, executive function, and ToM tasks was significantly predictive of time to conversion to psychosis. CONCLUSION: This study indicated that UHR patients who later converted to psychosis performed more poorly on tasks involving social cognition and neurocognition than did those who did not convert. We suggest that these deficits can serve as specific markers to predict the development of psychosis. PMID- 21620682 TI - Assessing everyday functioning in schizophrenia: not all informants seem equally informative. AB - Self reports of everyday functioning on the part of people with schizophrenia have been found to be poorly correlated with the reports of other informants and with their own performance of tests of cognition and functional abilities. However, it is not clear which informants are best for providing accurate reports of everyday functioning. This study examined the convergence between self-reports on the part of people with schizophrenia (n=193), whose real-world functioning was rated by a friend or relative (n=154), or a high contact clinician (n=39) across 6 functional status rating scales. In addition, correlations between these reports and patient's performance on neuropsychological tests and a performance based measure of functional capacity were also calculated. For convergence between raters, friend or relative informants and patient reports were significantly correlated for 4/6 rating scales. For the smaller sample of clinician informants, the correlations were significant on 2/6 scales. In the analyses of convergence between patient performance scores and functioning ratings, only 1/12 correlations between patient report and performance were significant, while friend or relative reports also were only correlated with performance on one rating scale. In contrast, clinician reports of functioning were correlated with patients' functional capacity performance on 4/6 rating scales and with neuropsychological test performance on 2/6. High contact clinicians appear to generate ratings of everyday functioning that are more closely linked to patients' ability scores than friend or relative informants. Later analyses will determine if there are differences between friend or relative informants. PMID- 21620683 TI - The protein kinase CK2 inhibitor TBB mediates up-regulation of MEK3/6 and p38delta activities, down-regulation of ERK1/2 activity and induction of G1/S arrest in normal human epidermal autocrine proliferating keratinocytes. PMID- 21620684 TI - Differential modulation of stress-inflammation responses by plant polyphenols in cultured normal human keratinocytes and immortalized HaCaT cells. AB - BACKGROUND: Environmental and endogenous stresses to skin are considered causative reasons for skin cancers, premature ageing, and chronic inflammation. Screening of substances with preventive and/or curative properties is currently based on mechanistic studies of their effects towards stress-induced responses in skin cell cultures. OBJECTIVE: We compared effects of plant polyphenols (PPs) on the constitutive, UVA-, LPS-, or TNF-alpha-induced inflammatory responses in cultured normal human epidermal keratinocytes (NHEK) and immortalized HaCaT cells. METHODS: Representatives of three classes of PPs, flavonoids, stilbenoids, and phenylpropanoids were studied. Their effects on mRNA were determined by qRT PCR; protein expression was assayed by Western blot and bioplexed ELISA; phosphorylation of Akt1, ERK1/2, EGFR, and NFkappaB was quantified by intracellular ELISA or Western blot. RESULTS: PPs or their combination with UVA or LPS induced strong up-regulation of stress responses in HaCaT but not in NHEK. In addition, compared to NHEK, HaCaT responded to TNF-alpha with higher synthesis of MCP-1, IP-10 and IL-8, concomitant with stronger NFkappaB activation. PPs down regulated the chemokine release from both cell types, although with distinct effects on NFkappaB, Akt1, ERK, and EGFR activation. CONCLUSION: Results of pharmacological screenings obtained by using HaCaT should be cautiously considered while extending them to primary keratinocytes from human epidermis. PMID- 21620685 TI - SHARPIN regulates mitochondria-dependent apoptosis in keratinocytes. AB - BACKGROUND: The chronic proliferative dermatitis mutation (CPDM) in mice, due to Sharpin deficiency (Sharpin(cpdm)), is a multisystem disorder characterized by peripheral blood eosinophilia and eosinophil infiltration of affected tissues including the skin, bone marrow, spleen, lung, heart, and other organs. The epidermis has numerous apoptotic keratinocytes which increase with age, coalesce, form vesicles, and rupture causing ulceration. OBJECTIVE: To clarify the molecular pathways involved in the keratinocyte apoptosis caused by loss of function of SHARPIN in mice. METHOD: 10-week-old Sharpin(cpdm) and wildtype mice were used for experiments. Ultrastructural changes of skin were evaluated by transmission electron microscopy. Cross points of mitochondrial pathway were analyzed by in vitro and in vivo cellular and molecular assays. RESULTS: 77.5% skin cells in Sharpin(cpdm) mice were functionally apoptotic and dead cells, compared to only 18.1% unhealthy skin cells in wildtype mice, indicated by annexin-V/propidium iodide FACS analysis. Mitochondria in keratinocytes were disrupted containing prominent electron dense inclusions and membrane potential depolarization, accompanied by a shift in protein expression between the anti apoptotic BCL2 and pro-apoptotic BAX proteins. Enzymatic activities of caspases 9 and 3, but not 8, were markedly increased in Sharpin(cpdm) keratinocytes. Caspase 3 was cleaved in most cells in skin of 10-week-old mutant mice. CONCLUSION: The present results indicated that keratinocyte apoptosis in Sharpin(cpdm) mice was regulated by an intrinsic caspase-dependent mitochondria pathway. PMID- 21620686 TI - Molecular simulations predict novel collagen conformations during cross-link loading. AB - Collagen cross-linking mechanically strengthens tissues during development and aging, but there is limited data describing how force transmitted across cross links affects molecular conformation. We used Steered Molecular Dynamics (SMD) to model perpendicular force through a side chain. Results predicted that collagen peptides have negligible bending resistance and that mechanical force causes helix disruption below covalent bond failure strength, suggesting alternative molecular conformations precede cross-link rupture and macroscopic damage during mechanical loading. PMID- 21620687 TI - Alteration of municipal solid waste incineration bottom ash focusing on the evolution of iron-rich constituents. AB - Municipal solid waste incineration (MSWI) bottom ash contains a considerable amount of Fe-rich constituents. The behaviors of these constituents, such as dissolution and precipitation, are quite important as they regulate the distribution of a series of ions between the liquid (percolated fluid) and solid (ash deposit) phases. This paper studied both fresh and weathered MSWI bottom ash from the mineralogical and geochemical viewpoint by utilizing optical microscopy, scanning electron microscopy/energy dispersive X-ray spectroscopy (SEM/EDX), and powder X-ray diffraction. The analysis results revealed that for the fresh bottom ash, iron preferentially existed in the chemical forms of spinel group (mainly Fe(3)O(4), and a series of Al- or Ti- substituted varieties), metallic inclusions (including Fe-P, Fe-S, Fe-Cu-Pb), hematite (Fe(2)O(3)) and unburned iron pieces. In the 1-20 years weathered bottom ash collected from a landfill site, interconversions among these Fe-rich constituents were identified. Consequently, numerous secondary products were developed, including goethite (alpha-FeOOH), lepidocrocite (gamma-FeOOH), hematite, magnetite, wustite (FeO), Fe-Si-rich gel phase. Of all these transformation products, hydrous iron oxides were the most common secondary minerals. Quantitative chemical analysis of these secondary products by SEM/EDX disclosed a strong association between the newly formed hydrous iron oxides and heavy metals (e.g. Pb, Zn, Ni, and Cu). The results of this study suggest that the processes of natural weathering and secondary mineralization contribute to reduction of the potential risks of heavy metals to the surrounding environments. PMID- 21620688 TI - Metabolic engineering of microbial pathways for advanced biofuels production. AB - Production of biofuels from renewable resources such as cellulosic biomass provides a source of liquid transportation fuel to replace petroleum-based fuels. This endeavor requires the conversion of cellulosic biomass into simple sugars, and the conversion of simple sugars into biofuels. Recently, microorganisms have been engineered to convert simple sugars into several types of biofuels, such as alcohols, fatty acid alkyl esters, alkanes, and terpenes, with high titers and yields. Here, we review recently engineered biosynthetic pathways from the well characterized microorganisms Escherichia coli and Saccharomyces cerevisiae for the production of several advanced biofuels. PMID- 21620689 TI - Hormonal and reproductive factors in relation to melanoma in women: current review and meta-analysis. AB - A number of studies have focused on possible relationships between characteristics of female endocrine status and melanoma (CM) risk; however, the link between melanoma, oral contraceptive (OC) and hormonal replacement therapy (HRT) use, and reproductive factors remains controversial. A comprehensive, systematic bibliographic search of the medical literature was conducted to identify relevant studies. Random effects models were used to summarise results. Subgroup, meta-regression and sensitivity analyses have been carried out to explore sources of between-study variation and bias. We included thirty-six observational studies published in the last 30 years. Summarising a total of 5626 melanoma cases, we did not find any significant melanoma risk associated with OC and HRT use. Several reproductive factors were also investigated, summarising data on 16787 melanoma cases. We found a significantly increased melanoma risk for late age at first birth, and women with more than one child may be at a lower risk for melanoma; however, socio-economic confounders were found to play a significant role in explaining this association. This study confirmed no increased risk of CM with the use of oral contraceptives and hormone replacement therapy: exogenous female hormones do not contribute to an increased risk of CM. In contrast, significant associations of CM with parity and age at first pregnancy were observed in this meta-analysis finds and warrant further research. PMID- 21620690 TI - Acid-catalyzed conversion of xylose, xylan and straw into furfural by microwave assisted reaction. AB - Furfural is a biomass derived-chemical that can be used to replace petrochemicals. In this study, the acid-catalyzed conversion of xylose and xylan to furfural by microwave-assisted reaction was investigated at selected ranges of temperature (140-190 degrees C), time (1-30 min), substrate concentration (1:5 1:200 solid:liquid ratio), and pH (2-0.13). We found that a temperature of 180 degrees C, a solid:liquid ratio of 1:200, a residence time of 20 min, and a pH of 1.12 gave the best furfural yields. The effect of different Bronsted acids on the conversion efficiency of xylose and xylan was also evaluated, with hydrochloric acid being found to be the most effective catalyst. The microwave-assisted process provides highly efficient conversion: furfural yields obtained from wheat straw, triticale straw, and flax shives were 48.4%, 45.7%, and 72.1%, respectively. PMID- 21620691 TI - Transition of microbial communities during the adaption to anaerobic digestion of carrot waste. AB - In this study a microbial community suitable for anaerobic digestion of carrot pomace was developed from inocula obtained from natural environmental sources. The changes along the process were monitored using pyrosequencing of the 16S rRNA gene. As the community adapted from a diverse natural community to a community with a definite function, diversity decreased drastically. Major bacterial groups remaining after enrichment were Bacilli (31-45.3%), Porphyromonadaceae (12.1 24.8%) and Spirochaetes (12.5-18.5%). The archaeal population was even less diverse and mainly represented by a single OTU that was 99.7% similar to Methanosarcina mazei. One enrichment which failed to produce large amounts of methane had shifts in the bacterial populations and loss of methanogenic archaea. PMID- 21620692 TI - Citric acid modified kenaf core fibres for removal of methylene blue from aqueous solution. AB - Chemically modified kenaf core fibres were prepared via esterification in the presence of citric acid (CA). The adsorption kinetics and isotherm studies were carried out under different conditions to examine the adsorption efficiency of CA treated kenaf core fibres towards methylene blue (MB). The adsorption capacity of the kenaf core fibres increased significantly after the citric acid treatment. The values of the correlation coefficients indicated that the Langmuir isotherm fitted the experimental data better than the Freundlich isotherm. The maximum adsorption capacity of the CA-treated kenaf core fibres was found to be 131.6mg/g at 60 degrees C. Kinetic models, pseudo-first-order, pseudo-second-order and intraparticle diffusion, were employed to describe the adsorption mechanism. The kinetic data were found to fit pseudo-second-order model equation as compared to pseudo-first-order model. The adsorption of MB onto the CA-treated kenaf core fibres was spontaneous and endothermic. PMID- 21620693 TI - Performance of a mobile mechanical screen to improve the commercial quality of wood chips for energy. AB - The study analyzed the performance of a mobile screening device for upgrading coarse wood chips to residential user standards, by removing oversize particles and fines. The machine was designed for transportation to forest landings, logistic terminals and plant chip yards. Average productivity was 1.9 oven-dry tons (odt) h(-1), corresponding to a screening cost of 28.5? odt(-1). This figure was lower than the price increase obtained by upgrading industrial chips to residential user standards. Hence, screening offered a profit of 4.7? odt(-1), or 16% of the original screening cost. The screening process was capable of upgrading chips from industrial to residential specifications, by reducing the incidence of oversize particles below the 1% critical threshold. Screening also allowed a substantial reduction in the content of fines. A similar effect was not verified for crushed wood, which failed to meet the specifications for residential fuel. PMID- 21620694 TI - Aerobic denitrification by novel isolated strain using NO-2-N as nitrogen source. AB - Biological denitrification reaction can be achieved under aerobic environment. Few aerobic denitrifiers using nitrite as sole nitrogen source were identified. Using nitrite as the sole nitrogen source, this work assessed the denitrification activity of yy7, an aerobic heterotrophic denitrifier identified as Pseudomonas sp. (94% similarity) by 16S rRNA sequencing analysis. The logistic equation describes the cell growth curve, yielding a generation time of 2.9h at an initial 18 mg l(-1)NO(-)2-N. Reduction of NO(-)2-N was primarily achieved during its logarithmic growth phase, and was accompanied by an increase in suspension pH and near complete consumption of dissolved oxygen. Three genes relating to nirK, norB, and nosZ were noted to involve in isolate strain. Isolate yy7 can survive and remove up to 40 mg l(-1)NO(-)2-N and, hence, can be applied as an effective aerobic denitrifier during simultaneous nitrification and denitrification via nitrite processes. PMID- 21620695 TI - Characterization of organic matter degradation during composting of manure-straw mixtures spiked with tetracyclines. AB - The objective of this study was to investigate humification and mineralization of manure-straw mixtures contaminated by tetracyclines during composting. Hen manure, pig manure and rice straw were used as the raw materials. The manure straw mixtures were spiked with tetracycline, chlortetracycline, and oxytetracycline at the concentration of 60 mg/kg dry matter. The results show that tetracyclines had no obvious influence on the composting process and more than 93% of the tetracyclines was decreased during a 45-day composting. The Fourier transform infrared (FTIR) spectra indicated that easily biodegradable components such as aliphatic substrates, carbohydrates and polysaccharides were decomposed and the contents of aromatic components relatively rose during the composting. The X-ray diffraction (XRD) spectra confirmed the natural formation of struvite, the degradation of easily biodegradable components, and the mineralization of organic matter during the composting. Therefore, FTIR and XRD analysis can be useful tools for monitoring the composting process. PMID- 21620696 TI - Modified GM3 gangliosides produced by metabolic oligosaccharide engineering. AB - Metabolic oligosaccharide engineering is powerful approach to altering the structure of cellular sialosides. This method relies on culturing cells with N acetylmannosamine (ManNAc) analogs that are metabolized to their sialic acid counterparts and added to glycoproteins and glycolipids. Here we employed two cell lines that are deficient in ManNAc biosynthesis and examined their relative abilities to metabolize a panel of ManNAc analogs to sialosides. In addition to measuring global sialoside production, we also examined biosynthesis of the sialic acid-containing glycolipid, GM3. We discovered that the two cell lines differ in their ability to discriminate among the variant forms of ManNAc. Further, our data suggest that modified forms of sialic acid may be preferentially incorporated into certain sialosides and excluded from others. Taken together, our results demonstrate that global analysis of sialoside production can obscure sialoside-specific differences. These findings have implications for downstream applications of metabolic oligosaccharide engineering, including imaging and proteomics. PMID- 21620697 TI - Synthesis and structure-activity relationships of nitrobenzyl phosphoramide mustards as nitroreductase-activated prodrugs. AB - A series of nitrobenzyl phosphoramide mustards and their analogs was designed and synthesized to explore their structure-activity relationships as substrates of nitroreductases from Escherichia coli and trypanosomes and as potential antiproliferative and antiparasitic agents. The position of the nitro group on the phenyl ring was important with the 4-nitrobenzyl phosphoramide mustard (1) offering the best combination of enzyme activity and antiproliferative effect against both mammalian and trypanosomatid cells. A preference was observed for halogen substitutions ortho to benzyl phosphoramide mustard but distinct differences were found in their SAR of substituted 4-nitrobenzyl phosphoramide mustards in E. coli nitroreductase-expressing cells and in trypanosomatids expressing endogenous nitroreductases. PMID- 21620698 TI - Discovery of novel xanthone derivatives as xanthine oxidase inhibitors. AB - Xanthine oxidase is the key enzyme that catalyzes the oxidation of hypoxanthine to xanthine and then to uric acid. In this study, a series of xanthone derivatives were synthesized as effective and a new class of xanthine oxidase inhibitor. Compounds 8a, 8c, 8i, 8g and 8r showed good inhibition against xanthine oxidase. The presence of a cyano group at the para position of benzyl moiety turned out to be the preferred substitution pattern. Molecular modeling studies were performed to gain an insight into its binding mode with xanthine oxidase, and to provide the basis for further structure-guided design of new non purine xanthine oxidase inhibitors associated with the xanthone framework. PMID- 21620699 TI - Substituted N-aryl-6-pyrimidinones: a new class of potent, selective, and orally active p38 MAP kinase inhibitors. AB - A novel series of highly potent and selective p38 MAP kinase inhibitors was developed originating from a substituted N-aryl-6-pyrimidinone scaffold. SAR studies coupled with in vivo evaluations in rat arthritis model culminated in the identification of 10 with excellent oral efficacy. Compound 10 exhibited a significantly enhanced dissolution rate compared to 1, translating to a high oral bioavailability (>90%) in rat. In animal studies 10 inhibited LPS-stimulated production of tumor necrosis factor-alpha in a dose-dependent manner and demonstrated robust efficacy comparable to dexamethasone in a rat streptococcal cell wall-induced arthritis model. PMID- 21620700 TI - Chemical genetics reveals negative regulation of abscisic acid signaling by a plant immune response pathway. AB - Coordinated regulation of protection mechanisms against environmental abiotic stress and pathogen attack is essential for plant adaptation and survival. Initial abiotic stress can interfere with disease-resistance signaling [1-6]. Conversely, initial plant immune signaling may interrupt subsequent abscisic acid (ABA) signal transduction [7, 8]. However, the processes involved in this crosstalk between these signaling networks have not been determined. By screening a 9600-compound chemical library, we identified a small molecule [5-(3,4 dichlorophenyl)furan-2-yl]-piperidine-1-ylmethanethione (DFPM) that rapidly downregulates ABA-dependent gene expression and also inhibits ABA-induced stomatal closure. Transcriptome analyses show that DFPM also stimulates expression of plant defense-related genes. Major early regulators of pathogen resistance responses, including EDS1, PAD4, RAR1, and SGT1b, are required for DFPM-and notably also for Pseudomonas-interference with ABA signal transduction, whereas salicylic acid, EDS16, and NPR1 are not necessary. Although DFPM does not interfere with early ABA perception by PYR/RCAR receptors or ABA activation of SnRK2 kinases, it disrupts cytosolic Ca(2+) signaling and downstream anion channel activation in a PAD4-dependent manner. Our findings provide evidence that activation of EDS1/PAD4-dependent plant immune responses rapidly disrupts ABA signal transduction and that this occurs at the level of Ca(2+) signaling, illuminating how the initial biotic stress pathway interferes with ABA signaling. PMID- 21620701 TI - A molecular switch for initiating cell differentiation in Arabidopsis. AB - BACKGROUND: The onset of differentiation entails modifying the gene expression state of cells, to allow activation of developmental programs that are maintained repressed in the undifferentiated precursor cells [1, 2]. This requires a mechanism to change gene expression on a genome-scale. Recent evidence suggests that in mammalian stem cells, derepression of developmental regulators during differentiation involves a shift from stalled to productive elongation of their transcripts [3-5], but factors mediating this shift have not been identified and the evidence remains correlative. RESULTS: We report the identification of the MINIYO (IYO) gene, a positive regulator of transcriptional elongation that is essential for cells to initiate differentiation in Arabidopsis. IYO interacts with RNA polymerase II and the Elongator complex and is required to sustain global levels of transcriptional elongation activity, specifically in differentiating tissues. Accordingly, IYO is expressed in embryos, meristems, and organ primordia and not in mature tissues. Moreover, differential subcellular protein distribution further refines the domain of IYO function by directing nuclear accumulation, and thus its transcriptional activity, to cells initiating differentiation. Importantly, IYO overexpression induces premature cell differentiation and leads to meristem termination phenotypes. CONCLUSIONS: These findings identify IYO as a necessary and sufficient factor for initiating differentiation in Arabidopsis and suggest that the targeted nuclear accumulation of IYO functions as a transcriptional switch for this fate transition. PMID- 21620702 TI - A mutually inhibitory interaction between auxin and cytokinin specifies vascular pattern in roots. AB - BACKGROUND: Whereas the majority of animals develop toward a predetermined body plan, plants show iterative growth and continually produce new organs and structures from actively dividing meristems. This raises an intriguing question: How are these newly developed organs patterned? In Arabidopsis embryos, radial symmetry is broken by the bisymmetric specification of the cotyledons in the apical domain. Subsequently, this bisymmetry is propagated to the root promeristem. RESULTS: Here we present a mutually inhibitory feedback loop between auxin and cytokinin that sets distinct boundaries of hormonal output. Cytokinins promote the bisymmetric distribution of the PIN-FORMED (PIN) auxin efflux proteins, which channel auxin toward a central domain. High auxin promotes transcription of the cytokinin signaling inhibitor AHP6, which closes the interaction loop. This bisymmetric auxin response domain specifies the differentiation of protoxylem in a bisymmetric pattern. In embryonic roots, cytokinin is required to translate a bisymmetric auxin response in the cotyledons to a bisymmetric vascular pattern in the root promeristem. CONCLUSIONS: Our results present an interactive feedback loop between hormonal signaling and transport by which small biases in hormonal input are propagated into distinct signaling domains to specify the vascular pattern in the root meristem. It is an intriguing possibility that such a mechanism could transform radial patterns and allow continuous vascular connections between other newly emerging organs. PMID- 21620703 TI - Spire-type actin nucleators cooperate with Formin-2 to drive asymmetric oocyte division. AB - Oocytes mature into eggs by extruding half of their chromosomes in a small cell termed the polar body. Asymmetric oocyte division is essential for fertility [1], but despite its importance, little is known about its mechanism. In mammals, the meiotic spindle initially forms close to the center of the oocyte. Thus, two steps are required for asymmetric meiotic division: first, asymmetric spindle positioning and second, polar body extrusion. Here, we identify Spire1 and Spire2 as new key factors in asymmetric division of mouse oocytes. Spire proteins are novel types of actin nucleators that drive nucleation of actin filaments with their four WH2 actin-binding domains [2-6]. We show that Spire1 and Spire2 first mediate asymmetric spindle positioning by assembling an actin network that serves as a substrate for spindle movement. Second, they drive polar body extrusion by promoting assembly of the cleavage furrow. Our data suggest that Spire1 and Spire2 cooperate with Formin-2 (Fmn2) to nucleate actin filaments in mouse oocytes and that both types of nucleators act as a functional unit. This study not only reveals how Spire1 and Spire2 drive two critical steps of asymmetric oocyte division, but it also uncovers the first physiological function of Spire type actin nucleators in vertebrates. PMID- 21620704 TI - Characterization of dip1p reveals a switch in Arp2/3-dependent actin assembly for fission yeast endocytosis. AB - BACKGROUND: During endocytosis in yeast, a choreographed series of discrete local events at the plasma membrane lead to a rapid burst of actin polymerization and the subsequent internalization of an endocytic vesicle. What initiates Arp2/3 dependent actin polymerization in this process is not well understood. RESULTS: The Schizosaccharomyces pombe WISH/DIP/SPIN90 ortholog dip1p is an actin-patch protein that regulates the temporal sequence of endocytic events. dip1Delta mutants exhibit a novel phenotype in which early events such as WASp localization occur normally but arrival of Arp2/3, actin polymerization, and subsequent steps are delayed and occur with apparently random timing. In studying this mutant, we demonstrate that positive feedback loops of WASp, rapid actin assembly, and Arp2/3 contribute to switch-like behavior that initiates actin polymerization. In the absence of dip1p, a subset of patches is activated concurrently with the "touch" of a neighboring endocytic vesicle. CONCLUSIONS: These studies reveal a switch-like mechanism responsible for the initiation of actin assembly during endocytosis. This switch may be activated in at least two ways, through a dip1p dependent mechanism and through contact with another endocytic vesicle. PMID- 21620705 TI - Phloem-transported cytokinin regulates polar auxin transport and maintains vascular pattern in the root meristem. AB - Cytokinin phytohormones regulate a variety of developmental processes in the root such as meristem size, vascular pattern, and root architecture [1-3]. Long distance transport of cytokinin is supported by the discovery of cytokinins in xylem and phloem sap [4] and by grafting experiments between wild-type and cytokinin biosynthesis mutants [5]. Acropetal transport of cytokinin (toward the shoot apex) has also been implicated in the control of shoot branching [6]. However, neither the mode of transport nor a developmental role has been shown for basipetal transport of cytokinin (toward the root apex). In this paper, we combine the use of a new technology that blocks symplastic connections in the phloem with a novel approach to visualize radiolabeled hormones in planta to examine the basipetal transport of cytokinin. We show that this occurs through symplastic connections in the phloem. The reduction of cytokinin levels in the phloem leads to a destabilization of the root vascular pattern in a manner similar to mutants affected in auxin transport or cytokinin signaling [7]. Together, our results demonstrate a role for long-distance basipetal transport of cytokinin in controlling polar auxin transport and maintaining the vascular pattern in the root meristem. PMID- 21620706 TI - Causal role of dorsolateral prefrontal cortex in human perceptual decision making. AB - The way that we interpret and interact with the world entails making decisions on the basis of available sensory evidence. Recent primate neurophysiology [1-6], human neuroimaging [7-13], and modeling experiments [14-19] have demonstrated that perceptual decisions are based on an integrative process in which sensory evidence accumulates over time until an internal decision bound is reached. Here we used repetitive transcranial magnetic stimulation (rTMS) to provide causal support for the role of the dorsolateral prefrontal cortex (DLPFC) in this integrative process. Specifically, we used a speeded perceptual categorization task designed to induce a time-dependent accumulation of sensory evidence through rapidly updating dynamic stimuli and found that disruption of the left DLPFC with low-frequency rTMS reduced accuracy and increased response times relative to a sham condition. Importantly, using the drift-diffusion model, we show that these behavioral effects correspond to a decrease in drift rate, a parameter describing the rate and thereby the efficiency of the sensory evidence integration in the decision process. These results provide causal evidence linking the DLPFC to the mechanism of evidence accumulation during perceptual decision making. PMID- 21620707 TI - Different roles for homologous interneurons in species exhibiting similar rhythmic behaviors. AB - It is often assumed that similar behaviors in related species are produced by similar neural mechanisms. To test this, we examined the neuronal basis of a simple swimming behavior in two nudibranchs (Mollusca, Opisthobranchia), Melibe leonina and Dendronotus iris. The side-to-side swimming movements of Dendronotus [1] strongly resemble those of Melibe [2, 3]. In Melibe, it was previously shown that the central pattern generator (CPG) for swimming is composed of two bilaterally symmetric pairs of identified interneurons, swim interneuron 1 (Si1) and swim interneuron 2 (Si2), which are electrically coupled ipsilaterally and mutually inhibit both contralateral counterparts [2, 4]. We identified homologs of Si1 and Si2 in Dendronotus. (Henceforth, homologous neurons in each species will be distinguished by the subscripts (Den) and (Mel).) We found that Si2(Den) and Si2(Mel) play similar roles in generating the swim motor pattern. However, unlike Si1(Mel), Si1(Den) was not part of the swim CPG, was not strongly coupled to the ipsilateral Si2(Den), and did not inhibit the contralateral neurons. Thus, species differences exist in the neuronal organization of the swim CPGs despite the similarity of the behaviors. Therefore, similarity in species-typical behavior is not necessarily predictive of common neural mechanisms, even for homologous neurons in closely related species. PMID- 21620708 TI - Modality-independent coding of spatial layout in the human brain. AB - In many nonhuman species, neural computations of navigational information such as position and orientation are not tied to a specific sensory modality [1, 2]. Rather, spatial signals are integrated from multiple input sources, likely leading to abstract representations of space. In contrast, the potential for abstract spatial representations in humans is not known, because most neuroscientific experiments on human navigation have focused exclusively on visual cues. Here, we tested the modality independence hypothesis with two functional magnetic resonance imaging (fMRI) experiments that characterized computations in regions implicated in processing spatial layout [3]. According to the hypothesis, such regions should be recruited for spatial computation of 3D geometric configuration, independent of a specific sensory modality. In support of this view, sighted participants showed strong activation of the parahippocampal place area (PPA) and the retrosplenial cortex (RSC) for visual and haptic exploration of information-matched scenes but not objects. Functional connectivity analyses suggested that these effects were not related to visual recoding, which was further supported by a similar preference for haptic scenes found with blind participants. Taken together, these findings establish the PPA/RSC network as critical in modality-independent spatial computations and provide important evidence for a theory of high-level abstract spatial information processing in the human brain. PMID- 21620709 TI - Interventional treatment of carotid cavernous fistula. AB - We reviewed retrospectively the outcomes of interventional endovascular treatment of direct or dural (indirect) carotid cavernous fistulas in 24 consecutive patients requiring endovascular treatment at Royal Prince Alfred Hospital between 1994 and 2009. Data was collected from each patient's neurological, ophthalmological and radiological reports. Of the 12 patients with direct fistulas all had signs of orbital and ocular venous congestion and ophthalmoplegia; nine also had reduced vision ranging from 6/9 to nil perception of light, two had normal vision and one was unconscious. Nine of the 12 direct fistulas were embolized transarterially, two transvenously, one by a combination of both approaches and all were successfully closed. After treatment, seven of the nine patients with reduced vision had complete or nearly complete restoration of vision,while two who presented with nil perception of light (one in both eyes) had no recovery of vision. In contrast, seven of the 12 patients with dural fistulas had ophthalmoplegia, three had reduced vision, down to 6/24 and one did not have any sign of venous congestion. Vision recovered in all three patients after embolization of the dural fistula. Dural fistulas were embolized transvenously in 11 and transarterially in one patient. Apart from ophthalmoplegia, all other ocular signs and symptoms rapidly resolved after closure of the fistula in each of the 24 patients. The diagnosis was delayed by being missed either during the first admission or at the first specialist consultation in three of the 12 patients with direct fistulas, and in seven of the 12 patients with dural fistulas. One patient with a direct and another with a dural fistula had limited cerebral infarctions during embolization. In this series, endovascular interventional treatment of carotid cavenous fistulas restored visual loss in 10 of 12 patients with visual loss. The two who did not recover had presented with nil perception of light, one after a delay in diagnosis of 6 weeks. Some degree of ophthalmoplegia tended to remain. This emphasizes the need for early diagnosis and treatment before visual loss or ophthalmoplegia becomes severe. PMID- 21620710 TI - Dynamic protein-DNA recognition: beyond what can be seen. AB - Traditionally, specific DNA recognition is thought to rely on static contacts with the bases or phosphates. Recent results, however, indicate that residues far outside the binding context can crucially influence selectivity or binding affinity via transient, dynamic interactions with the DNA binding interface. These regions usually do not adopt a well-defined structure, even when bound to DNA, and thus form a fuzzy complex. Here, we propose the existence of a dynamic DNA readout mechanism, wherein distant segments modulate conformational preferences, flexibility or spacing of the DNA binding motifs or serve as competitive partners. Despite their low sequence similarity, these intrinsically disordered regions are often conserved at the structural level, and exploited for regulation of the transcription machinery via protein-protein interactions, post translational modifications or alternative splicing. PMID- 21620711 TI - Evaluation of early-stage osteochondral defect repair using a biphasic scaffold based on a collagen-glycosaminoglycan biopolymer in a caprine model. AB - The aim of this study was to evaluate a new collagen-GAG-calcium phosphate biphasic scaffold for the repair of surgically created osteochondral defects in goats. Comparison of morphological, histological and mechanical performance of the repair tissue was made with defects repaired using a synthetic polymer scaffold. Defects were created in the medial femoral condyle (MFC) and lateral trochlear sulcus (LTS) of Boer Cross goats and evaluated at 12 and 26 weeks. It was found that the total histology score of the collagen-GAG based biomaterial (23.8; SD 1.7) provided a significant improvement (p<0.05) over the biphasic PLGA material (19;3) and the empty control defect (17.3;1.2) in the LTS. The overall trajectory of histological and morphological improvement between 12 and 26 weeks was found to be higher for the collagen-GAG scaffold compared to the PLGA material. The occurrence of sub-chondral bone cysts was lower for the collagen GAG scaffold with an incidence of 17% of defects, compared to 67% for the PLGA material at 26 weeks. The cartilage repair tissue for both materials evaluated was superior after 26 weeks implantation than the empty control with 75% of the collagen-GAG-treated defects showing markedly more hyaline-like cartilage and 50% of the PLGA sites exhibiting hyaline-like appearances, compared to 17% for the empty control. These early stage data indicate biphasic scaffolds based on collagen-GAG and PLGA both provide indications of satisfactory development of a structural repair to surgically prepared osteochondral defects. Furthermore, the biomaterial composition of the collagen-GAG may provide a more favourable environment for osteochondral repair. PMID- 21620712 TI - DOTA derivatives for site-specific biomolecule-modification via click chemistry: synthesis and comparison of reaction characteristics. AB - Due to the high stability of its complexes with many M(2+) and M(3+)-ions, DOTA (1,4,7,10-tetraazacyclododecane-N,N',N",N'''-tetraacetic acid) is the most commonly used chelator for the derivatization and radiolabeling of bioactive molecules. Most of the currently used DOTA derivatives comprise amine-reactive functionalities, limiting their application to the derivatization of fully protected molecules or otherwise resulting in randomly distributed conjugation sites of undefined number. Click chemistry reactions are a valuable alternative to this unspecific conjugation as they proceed efficiently and chemoselectively under mild conditions allowing a site-specific derivatization of unprotected biomolecules. In this work, we describe straightforward syntheses of DOTA derivatives containing thiol, maleimide, aminooxy, aldehyde, alkyne, and azide functionalities, amenable to the currently most often used click chemistry reactions. Furthermore, the efficiency of the respective click reactions introducing DOTA into bioactive molecules was investigated. For each of the synthesized DOTA synthons, the site-specific and efficient conjugation to Tyr(3) octreotate could be shown. Among these, the addition and oxime formation reactions proceeded fast and without side reactions, giving the products in high yields of 64-83% after purification. The copper-catalyzed triazole formation reactions produced some side-products, giving the desired products in lower, but still reasonable overall yields of 19-25%. All synthesized peptide-DOTA conjugates were labeled with (68)Ga in high radiochemical yields of 96-99% and high specific activities providing compounds of high purity, demonstrating the applicability of all synthons for biomolecule modification and subsequent radiolabeling. PMID- 21620713 TI - Conformational variability of different sulfonamide inhibitors with thienyl acetamido moieties attributes to differential binding in the active site of cytosolic human carbonic anhydrase isoforms. AB - The X-ray crystal structures of the adducts of human carbonic anhydrase (hCA, EC 4.2.1.1) II complexed with two aromatic sulfonamides incorporating 2 thienylacetamido moieties are reported here. Although, the two inhibitors only differ by the presence of an additional 3-fluoro substituent on the 4-amino benzenesulfonamide scaffold, their inhibition profiles against the cytosolic isoforms hCA I, II, III, VII and XIII are quite different. These differences were rationalized based on the obtained X-ray crystal structures, and their comparison with other sulfonamide CA inhibitors with clinical applications, such as acetazolamide, methazolamide and dichlorophenamide. The conformations of the 2 thienylacetamido tails in the hCA II adducts of the two sulfonamides were highly different, although the benzenesulfonamide parts were superimposable. Specific interactions between structurally different inhibitors and amino acid residues present only in some considered isoforms have thus been evidenced. These findings can explain the high affinity of the 2-thienylacetamido benzenesulfonamides for some pharmacologically relevant CAs (i.e., isoforms II and VII) being also useful to design high affinity, more selective sulfonamide inhibitors of various CAs. PMID- 21620714 TI - Preliminary evaluation of a 3H imidazoquinoline library as dual TLR7/TLR8 antagonists. AB - Toll-like receptors (TLR) -7 and -8 are thought to play an important role in immune activation processes underlying the pathophysiology of HIV and several clinically important autoimmune diseases. Based on our earlier findings of TLR7 antagonistic activity in a 3H imidazoquinoline, we sought to examine a pilot library of 3H imidazoquinolines for dual TLR7/8 antagonists, since they remain a poorly explored chemotype. 2D-NOE experiments were employed to unequivocally characterize the compounds. A quinolinium compound 12, bearing p-methoxybenzyl substituents on N3 and N5 positions was identified as a lead. Compound 12 was found to inhibit both TLR7 and TLR8 at low micromolar concentrations. Our preliminary results suggest that alkylation with electron-rich substituents on the quinoline N5, or conversely, elimination of the fixed charge of the resultant quaternary amine on the quinolinium may yield more active compounds. PMID- 21620715 TI - Modified cellulose morphologies and its composites; SEM and TEM analysis. AB - The complex, multi-level super molecular architecture of cellulose has been the subject of interest for several decades. The mechanical, physical, and environmental properties of cellulose depend on the molecular, supramolecular and morphological structure of the cellulose. This paper gives a brief overview to micro structural analysis of cellulose, as studied using transmission electron microscopy and scanning electron microscopy. The application of these techniques to study the diverse morphology of cellulose and its composites is illustrated using several examples. PMID- 21620716 TI - The structure of neurexin 1alpha reveals features promoting a role as synaptic organizer. AB - alpha-neurexins are essential synaptic adhesion molecules implicated in autism spectrum disorder and schizophrenia. The alpha-neurexin extracellular domain consists of six LNS domains interspersed by three EGF-like repeats and interacts with many different proteins in the synaptic cleft. To understand how alpha neurexins might function as synaptic organizers, we solved the structure of the neurexin 1alpha extracellular domain (n1alpha) to 2.65 A. The L-shaped molecule can be divided into a flexible repeat I (LNS1-EGF-A-LNS2), a rigid horseshoe shaped repeat II (LNS3-EGF-B-LNS4) with structural similarity to so-called reelin repeats, and an extended repeat III (LNS5-EGF-B-LNS6) with controlled flexibility. A 2.95 A structure of n1alpha carrying splice insert SS#3 in LNS4 reveals that SS#3 protrudes as a loop and does not alter the rigid arrangement of repeat II. The global architecture imposed by conserved structural features enables alpha-neurexins to recruit and organize proteins in distinct and variable ways, influenced by splicing, thereby promoting synaptic function. PMID- 21620718 TI - Antitumor potential induction and free radicals production in melanoma cells by Boron Neutron Capture Therapy. AB - Antiproliferative and oxidative damage effects occurring in Boron Neutron Capture Therapy (BNCT) in normal fibroblasts and melanoma cell lines were analyzed. Melanoma cells and normal fibroblasts were treated with different concentrations of Boronophenylalanine and irradiated with thermal neutron flux. The cellular viability and the oxidative stress were determined. BNCT induced free radicals production and proliferative potential inhibition in melanoma cells. Therefore, this therapeutic technique could be considered efficient to inhibit growth of melanoma with minimal effects on normal tissues. PMID- 21620717 TI - The crystal structure of the alpha-neurexin-1 extracellular region reveals a hinge point for mediating synaptic adhesion and function. AB - alpha- and beta-neurexins (NRXNs) are transmembrane cell adhesion proteins that localize to presynaptic membranes in neurons and interact with the postsynaptic neuroligins (NLGNs). Their gene mutations are associated with the autism spectrum disorders. The extracellular region of alpha-NRXNs, containing nine independently folded domains, has structural complexity and unique functional characteristics, distinguishing it from the smaller beta-NRXNs. We have solved the X-ray crystal structure of seven contiguous domains of the alpha-NRXN-1 extracellular region at 3.0 A resolution. The structure reveals an arrangement where the N-terminal five domains adopt a more rigid linear conformation and the two C-terminal domains form a separate arm connected by a flexible hinge. In an extended conformation the molecule is suitably configured to accommodate a bound NLGN molecule, as supported by structural comparison and surface plasmon resonance. These studies provide the structural basis for a multifunctional synaptic adhesion complex mediated by alpha-NRXN-1. PMID- 21620719 TI - Overexpression of 2-cysteine peroxiredoxin enhances tolerance to methyl viologen mediated oxidative stress and high temperature in potato plants. AB - Oxidative stress is one of the major causative factors for injury to plants exposed to environmental stresses. Plants have developed diverse defense mechanisms for scavenging oxidative stress-inducing molecules. The antioxidative enzyme 2-cysteine peroxiredoxin (2-Cys Prx) removes peroxides and protects the photosynthetic membrane from oxidative damage. In this study, transgenic potato (Solanum tuberosum L. cv. Atlantic) expressing At2-Cys Prx under control of the oxidative stress-inducible SWPA2 promoter or enhanced CaMV 35S promoter (referred to as SP and EP plants, respectively) was generated using Agrobacterium-mediated transformation. The transgenic plants were tested for tolerance to stress. Following treatment with 3 MUM methyl viologen (MV), leaf discs from SP and EP plants showed approximately 33 and 15% less damage than non-transformed (NT) plants. When 300 MUM MV was sprayed onto whole plants, the photosynthetic activity of SP plants decreased by 25%, whereas that of NT plants decreased by 60%. In addition, SP plants showed enhanced tolerance to high temperature at 42 degrees C. After treatment at high temperature, the photosynthetic activity of SP plants decreased by about 7% compared to plants grown at 25 degrees C, whereas it declined by 31% in NT plants. These results indicate that transgenic potato can efficiently regulate oxidative stress from various environmental stresses via overexpression of At2-Cys Prx under control of the stress-inducible SWPA2 promoter. PMID- 21620720 TI - Proposal of new nodal classifications for non-small-cell lung cancer based on the number and ratio of metastatic lymph nodes. AB - OBJECTIVE: The study aimed to evaluate the effectiveness of two new nodal classifications based on the number of metastatic lymph nodes (LNs) or ratio of metastatic to examined LNs (LNR) in making a prognosis, compared with the current nodal classification based on the location of metastatic LNs. METHODS: We analyzed 651 non-small-cell lung cancer patients who had undergone complete resection with the removal of more than five LNs between 1986 and 2003, excluding preoperative treatment cases, and a Tis, T4, N3, and M1 status, along with limited resection and operative death cases. The cutoff numbers for each category in the two new nodal classifications (number of metastatic LNs (nN0-2): 0, 1-2, and >3, and LNR (rN0-2): 0, 1-12, and >12%) were defined so that the numbers corresponded with paired categories within the current nodal classification. RESULTS: The 5-year survival rate was 75.4% for patients with the N0 categories in all three classifications. The 5-year survival rates for patients with N1 and N2 categories were 52.2% and 42.6% according to the current nodal classification, 54.3% and 39.8% according to the number of metastatic LNs, and 58.8% and 35.0% according to the LNR, respectively. Although all three nodal classifications were independent prognostic factors along with the age and pathological T status, when the three nodal classifications were entered into multivariate analysis individually, the hazard ratio of rN2 was the highest, at 3.15, followed by that of nN2 at 2.96. CONCLUSIONS: The LNR followed by the number of metastatic LNs may be more effective prognostic indicators than the current nodal classification based on the location of metastatic LNs. For the future revision, the number of metastatic LNs and LNR should be evaluated as indicators for the nodal classification of lung cancer. PMID- 21620721 TI - Bicuspid aortic valve leaflet morphology in relation to aortic root morphology: a study of 300 patients undergoing open-heart surgery. AB - OBJECTIVE: There is an ongoing discussion regarding the mechanism of aortic dilatation in bicuspid aortic valve (BAV) disease, that is, is this a hemodynamic effect or related to an inborn weakness of the aortic wall? This study evaluated the possibility of BAV morphology being related to ascending aorta morphology as such a correlation would strengthen the idea that hemodynamic alterations cause the dilatation of the aorta. METHODS: The morphology of the ascending aorta of 300 patients admitted for aortic valve and/or ascending aorta disease was evaluated by echocardiography and related to the surgeon's inspection of the aortic valve. RESULTS: A tricuspid aortic valve (TAV), BAV, or unicuspid aortic valve (UAV) was present in 130, 160, and 10 patients, respectively. Ascending aortic aneurysm was more common in patients with BAV compared with TAV (36% and 12%, respectively; p < 0.001), while ectasia of the aorta was similarly common (8% in both groups). Aortic stenosis or regurgitation was equally distributed in TAV and BAV patients with normal aortas (p=0.82). When the aorta was dilated, aortic stenosis was predominantly associated with BAV (BAV 56%, TAV 4%; p < 0.001), while aortic regurgitation was more common in TAV (TAV 81%, BAV 29%; p<0.001). In BAV patients, fusion of the right- and left coronary cusp was predominant (74%) followed by right- and non-coronary cusp fusion (14%) and true BAV (fusion of the right- and left coronary cusp without remnant raphe; 11%) (p < 0.001). The relative distribution of ascending aortic aneurysm or ectasia was similar in all morphologically different BAV (p = 0.95). CONCLUSIONS: In our study population, >50% of the patients admitted for surgery had a bicuspid valve. Aortic aneurysm was more common in BAV than in TAV patients. Aortic stenosis and aortic regurgitation were equally common in TAV and BAV with normal aortic dimensions, while aortic regurgitation was predominant in TAV with dilated aortas and aortic stenosis in BAV with dilated aortas. Dilatation of the aorta was similarly distributed regardless of BAV leaflet morphology. These findings support the idea of an intrinsic mechanisms underlying dilatation of the aorta in BAV patients. PMID- 21620723 TI - Quantitative proton MR spectroscopy as a biomarker of tumor necrosis in the rabbit VX2 liver tumor. AB - PURPOSE: To compare metabolic magnetic resonance (MR) imaging findings (ie, quantification of tumor choline concentration) with percentage of necrosis on pathologic examination in rabbits bearing VX2 liver tumors. MATERIALS AND METHODS: VX2 tumors were implanted in the livers of 16 rabbits. MR imaging was performed with a 1.5-T MR scanner and extremity coil, and a hydrogen-1 ((1)H) proton MR spectroscopy ((1)H MRS) imaging protocol was used. Rabbits were euthanized immediately after imaging, and the tumor was harvested and sliced at 4 mm intervals in the axial plane. Choline concentration was calculated and was compared with the percentage of tumor necrosis on pathologic examination. RESULTS: Mean tumor size at pathologic examination was 16 mm (range, 12-22 mm). Mean percentage of necrosis at pathologic examination was 22% (range, 4%-44%). Choline concentration showed a relatively high inverse correlation with percentage of necrosis on pathologic examination, with an r value of 0.78 (P < .002). CONCLUSIONS: Choline concentration showed a relatively high inverse correlation with tumor necrosis on pathologic examination. Therefore, (1)H MRS may be useful to assess tumor necrosis. PMID- 21620722 TI - Moving to tolerance: clinical application of T regulatory cells. AB - Decreasing the incidence of chronic rejection and reducing the need for life-long immunosuppression remain important goals in clinical transplantation. In this article, we will review how regulatory T cells (Treg) came to be recognized as an attractive way to prevent or treat allograft rejection, the ways in which Treg can be manipulated or expanded in vivo, and the potential of in vitro expanded/generated Treg for cellular therapy. We will describe the first regulatory T cell therapies that have been or are in the process of being conducted in the clinic as well as the safety concerns of such therapies and how outcomes may be measured. PMID- 21620724 TI - Daily magnesium intake and hypermagnesemia in hemodialysis patients with chronic kidney disease. AB - OBJECTIVE: The aim of this study was to evaluate daily magnesium intake and the relation to its serum concentration in hemodialysis (HD) patients with chronic kidney disease (CKD). DESIGN: This is a prospective, open-label, cross-sectional clinical study analyzing daily magnesium intake based on nutritional questionnaire. PARTICIPANTS: A total of 101 HD patients with CKD were screened for hypermagnesemia. All patients with serum magnesium >1.5 mmol/L were asked to fill in the standard 3-day nutritional questionnaire. The control group consisted of twice as many randomly selected HD patients with serum magnesium concentration <1.5 mmol/L and 20 subjects with normal kidney function on usual diet. RESULTS: Mean (+/-standard deviation) serum magnesium concentration in HD patients was 1.32 +/- 0.18 mmol/L. Hypermagnesemia >1.5 mmol/L was found in 17 (16.8%) patients. There was no one case of severe hypermagnesemia (>2.0 mmol/L). The daily intake of magnesium was higher by 31.7% in the group with serum magnesium >1.5 mmol/L. Hypermagnesemia was observed in patients ingesting >281 mg of magnesium daily. In univariate analysis, there was a strong positive correlation between magnesium intake and serum concentration in the whole group (r = 0.870, P < .001). No correlation between Kt/V or residual diuresis and serum magnesium concentration was found. CONCLUSIONS: Magnesium consumption is the most important determinant of serum magnesium concentration in HD patients with CKD. Magnesium containing phosphate binders can be considered in the therapy of hyperphosphatemia in HD patients without hypermagnesemia. PMID- 21620726 TI - Interdialytic weight gain does not influence the nutrition of new hemodialysis patients. AB - OBJECTIVE: The aim of the present study was to assess the relationship between interdialytic weight gain (IDWG) and nutrition markers in hemodialysis (HD) patients, by means of repeated measures analysis. METHODS: The records of 255 patients, who had recently received conventional HD for a minimum of 1 year, were retrospectively reviewed. Nutrition markers, including serum albumin, serum phosphate, blood urea nitrogen, and creatinine, were recorded at monthly intervals and subjected to repeated measures analysis. RESULTS: Patients with higher IDWG/dry weight (IDWG%) (>5%) had significantly lower body mass index throughout the study. Repeated measures analysis of variance indicated no significant difference in these nutrition markers for patients with different IDWG%. At the end of the study, neither IDWG nor IDWG% were found to be associated with albumin or phosphate, on linear regression analysis. CONCLUSIONS: There was no evidence of better nutrition in new HD patients with higher IDWG%. Although increased intake is promoted as critical for improving nutritional status in HD patients, it may be inappropriate to focus solely on the benefits of higher IDWG%, which can also lead to the development of hypertension, left ventricular hypertrophy, and intradialytic hypotension. PMID- 21620725 TI - The effect of fluid intake on chronic kidney transplant failure: a pilot study. AB - OBJECTIVE: Transplant recipients are generally instructed to increase their daily fluid intake so as to preserve kidney function. However, studies supporting this hypothesis are lacking. SETTING: Prospective, randomized study at a tertiary care university hospital. PATIENTS: Patients with chronic kidney transplant failure. INTERVENTION: Assignment to normal fluid intake (NFI: 2 L/day) or high fluid intake (HFI: 4 L/day) for 12 months. MAIN OUTCOME MEASURE: The effect of fluid intake on the decrease in estimated glomerular filtration rate (eGFR) was estimated by a mixed-effects general linear model. The analysis was adjusted for the observation period, age, intake of angiotensin-converting enzyme inhibitors or angiotensin II type 1 receptor blockers, diuretics, and transplant duration. RESULTS: A total of 33 patients were randomized to NFI and 29 to HFI. After 12 months, the mean eGFR had decreased to a similar extent in both groups (NFI: 44 +/- 9 mL/min vs. 41 +/- 9 mL/min; HFI: 46 +/- 15 mL/min vs. 44 +/- 15 mL/min). In the multivariate analysis, only the observation period had a significant effect on the decrease in eGFR. Randomization to NFI or HFI nor any other variable was associated with kidney function. The association between urine volume and urine osmolality was lost after 12 months. CONCLUSIONS: Recommendation of higher fluid intake does not seem to improve chronic kidney transplant failure. However, the lack of association between urine osmolality and reported urine volume at a later stage implies a loss of adherence to fluid intake over time. PMID- 21620727 TI - Association between vascular calcification scores on plain radiographs and fatty acid contents of erythrocyte membrane in hemodialysis patients. AB - OBJECTIVE: Vascular calcification (VC) scores determined by using simple plain radiographic films are known to be associated with coronary artery disease and mortality in patients undergoing hemodialysis (HD). Omega-3 fatty acid (FA) has been shown to reduce ectopic calcifications in an animal model, and it has also been shown that erythrocyte membrane omega-3 FA content is an independent discriminator of coronary artery disease. The present study was designed to demonstrate relations between VC scores and erythrocyte membrane FA contents in patients undergoing HD. DESIGN: A cross-sectional study was carried out. SETTING: The study was carried out at an outpatient hemodialysis unit at Dong-A University Hospital, Busan, Republic of Korea. PATIENTS: A total of 31 patients undergoing HD were recruited. Patients with significant malnutrition, a short duration of dialysis (<12 months), a history of recent infection, malignancy, or liver disease were excluded. MAIN OUTCOME MEASURES: Plain radiographic films of the feet, hands, pelvis, and lateral lumbar spine were examined and VC scores were determined using previously reported methods. Erythrocyte membrane FA contents were analyzed by gas chromatography. RESULTS: The erythrocyte membrane contents of eicosapentaenoic acid and docosahexaenoic acid were not found to be related with VC on simple plain radiographic films. However, erythrocyte membrane contents of oleic acid and total monounsaturated FA (MUFA) were significantly higher in patients with significant VC scores. Furthermore, erythrocyte membrane contents of MUFA and oleic acid were found to be negatively associated with high density lipoprotein cholesterol level and positively associated with triglyceride level. CONCLUSION: Erythrocyte membrane contents of MUFA and oleic acid were found to be associated with VC scores determined using plain radiographs and with dyslipidemia in patients undergoing HD. PMID- 21620728 TI - Robot-assisted mitral valve repair. PMID- 21620729 TI - Ultrasound examination for the diagnosis of pneumonia associated with Acinetobacter baumannii in a cardiac surgery patient. PMID- 21620730 TI - Recent advances in chronic thromboembolic pulmonary hypertension. AB - Surgical excellence in pulmonary thromboendarterectomy (PTE) for chronic thromboembolic pulmonary hypertension (CTEPH) has begun to spread around the world. The perioperative mortality for this procedure is typically under 10%. The maximal benefit from PTE is derived in those patients who have a high proximal clot burden that is surgically accessible, as outlined by the Jamieson classification. Residual pulmonary hypertension after successful PTE is common and increasingly is managed with maintenance oral pulmonary vasodilator therapy such as endothelin antagonists, phosphodiesterase inhibitors, and/or prostaglandins. The role of pulmonary vasodilator therapy in CTEPH before PTE is limited and should not delay definitive surgical therapy. Although plain deep hypothermic circulatory arrest (DHCA) is the classic technique for CTEPH, alternatives such as DHCA with antegrade cerebral perfusion are feasible as well. Prolonged mechanical ventilation after PTE remains common in part because of reperfusion pulmonary edema. Careful perioperative management can reduce the incidence of this syndrome. Because ventilator-associated pneumonia is also a common complication after PTE, it represents a major opportunity for outcome improvement, particularly because there are multiple modalities for its prevention and prompt diagnosis. PMID- 21620731 TI - Anterior mitral leaflet injury after debridement? PMID- 21620732 TI - Pulmonary hypertension and right ventricular dysfunction: physiology and perioperative management. PMID- 21620733 TI - Vehicle safety features, especially airbags, may account for the recent decline in heart donors. PMID- 21620734 TI - Heat shock protein 90 mediates anti-apoptotic effect of diazoxide by preventing the cleavage of Bid in hypothermic preservation rat hearts. AB - BACKGROUND: Successful organ preservation is the premise for clinical organ transplantation. The present study investigated whether heat shock protein 90 (Hsp90) is important in the anti-apoptotic effect of diazoxide in hypothermic preservation rat hearts. METHODS: Isolated rat hearts were preserved in Celsior solution, with or without diazoxide, for 3 to 9 hours, followed by 60 minutes of reperfusion. Cell apoptosis was assessed by terminal deoxynucleotide transferase mediated deoxy uridine triphosphate nick-end labeling. The left ventricular developed pressure (LVDP) was recorded. Expression of Hsp90 protein and cleavage of Bid were detected by Western blot and polymerase chain reaction. RESULTS: After hypothermic preservation for 3 to 9 hours, the LVDP recovery rate significantly decreased and cardiomyocyte apoptosis index increased in a time dependent manner. When compared with the 9-hour preservation group, Celsior solution supplemented with diazoxide significantly enhanced the LVDP recovery rate and decreased the apoptosis index. The cleavage of Bid increased after 9 hours of hypothermic preservation, which was inhibited by Celsior solution supplemented with diazoxide. Hypothermic preservation of rat hearts for 9 hours decreased the expression of Hsp90, whereas diazoxide supplementation significantly increased the expression of Hsp90. The Hsp90 inhibitor 17 allylamino-17-demethoxy-geldanamycin inhibited the diazoxide-induced decrease in cleavage of Bid, improvement of cardiac function, and decrease of apoptosis. Hsp90 inhibitor had no effect on the diazoxide-induced increase of total Cx43 protein expression in hearts preserved 9 hours, but inhibited the diazoxide induced increase of mitochondrial Cx43 protein level. CONCLUSION: Hsp90 might mediate diazoxide-induced cardioprotection against apoptosis in hypothermic preservation heart by preventing the cleavage of Bid. PMID- 21620735 TI - Heart transplantation and long-term mechanical support in Japan: progress at last. PMID- 21620736 TI - Lung transplantation in children with idiopathic pulmonary arterial hypertension: an 18-year experience. AB - BACKGROUND: The natural history of idiopathic pulmonary arterial hypertension (IPAH) in patients of all ages is one of relentless progression. For those who fail medical therapy, lung transplantation remains the ultimate palliation. In the USA, IPAH is the second leading indication for lung transplantation in children and first for children 1 to 5 years of age. In this study, we report our 18-year experience with lung transplantation in children with IPAH. METHODS: We performed a retrospective chart review of children with IPAH listed for lung transplant at our center between 1991 and 2009. Our data reflect a total of 26 children ranging in age from 1.6 to 18.9 years. Nineteen were transplanted and 7 died while waiting (27%). The impact of a number of pre-transplant variables on survival was evaluated. RESULTS: Median survival for those transplanted was 5.8 years, with 1- and 5-year survival rates of 95% and 61%, respectively. Survival was independent of pre-transplant considerations such as age, weight, need for intravenous (IV) inotropes, use of IV pulmonary vasodilators, year of transplant and severity of right-sided cardiac pressures. There was 1 hospital death. Compared with the transplanted group, children who died waiting had a significantly higher incidence of supra-systemic right heart pressures (p = 0.02) and hemoptysis (p = 0.01). CONCLUSIONS: Our study is the largest to date to look at outcomes for lung transplantation in children with IPAH. Their median survival compares favorably with that of all pediatric lung transplant recipients, 5.8 years vs 4.5 years, respectively. We did not identify any pre-transplant variables that presaged a poorer outcome. Thus, survival seemed more related to factors that influence long-term outcomes in all transplant recipients such as rejection and infection. Lung transplantation remains a viable option for children with IPAH, especially for those with supra-systemic right heart pressures despite maximal medical therapy. PMID- 21620737 TI - Severe acute kidney injury according to the RIFLE (risk, injury, failure, loss, end stage) criteria affects mortality in lung transplantation. AB - BACKGROUND: The RIFLE criteria (risk, injury, failure, loss, end stage) are new consensus definitions for acute kidney injury (AKI) associated with increased mortality; however, they have not been applied in lung transplantation (LTx). Using the RIFLE criteria, we examined the effect of AKI on outcomes and cost in LTx. METHODS: We retrospectively reviewed all LTx patients at our institution since the lung allocation score (LAS) system was initiated (May 2005-August 2010). Using the Modification of Diet in Renal Disease formula, we assigned appropriate RIFLE class (R, I, F) comparing baseline creatinine to peak levels in the first 7 days after LTx. Generalized linear models assessed the effect of AKI on in-hospital and 1-year mortality. Hospital charges were used to examine the financial effect of AKI. RESULTS: During the study, 106 LTx were performed. Excluding patients bridged to LTx with extracorporeal membrane oxygenation, 84 (86%) lived 1 year. Median LAS was 37.1 (interquartile range, 34.1-45.2). RIFLE status was I or F in 39 (36.7%), and 14 (13.2%) required renal replacement therapy (RRT). After adjusting for LAS, RIFLE-F had an increased relative rate (RR) of in-hospital mortality (RR, 4.76, 95% confidence interval [CI], 1.65-13.7, p = 0.004) and 1-year mortality (RR, 3.17, 95% CI 1.55-6.49, p = 0.002). RIFLE-R and I were not associated with higher in-hospital or 1-year mortality. Post operative RRT was associated with increased in-hospital (RR, 28.2; 95% CI, 6.18 128.1; p < 0.001) and 1-year mortality (RR, 4.97; 95% CI, 1.54-16.0; p < 0.001). AKI patients had higher median hospital charges of $168,146 vs $143,551 for no AKI (p = 0.02). CONCLUSIONS: This study shows high rates of AKI using the new RIFLE criteria in LTx. RIFLE-F is associated with higher in-hospital and 1-year mortality. Less severe degrees of AKI are not associated with increased mortality. The financial burden associated with AKI is significant. PMID- 21620738 TI - Vascular endothelial growth factor-D is overexpressed in human cardiac allograft vasculopathy and diabetic atherosclerosis and induces endothelial permeability to low-density lipoproteins in vitro. AB - BACKGROUND: Vascular endothelial growth factor (VEGF)-D is a member of the VEGF family, which can induce angiogenesis and lymphangiogenesis. We have previously demonstrated a role for VEGF-A in cardiac allograft vasculopathy (CAV). Our experiments profile the expression and localization of VEGF-D in human native atherosclerosis (NA), diabetes mellitus with atherosclerosis (DM) and CAV, and we investigate its ability to induce low-density lipoprotein (LDL) permeability in human cardiac microvascular endothelial cells (HCMEC). METHODS: VEGF-D mRNA and protein expression was characterized in coronary arteries and intramyocardial arterioles in NA, DM and CAV using in situ hybridization and immunohistochemical staining. Transendothelial electrical resistance (TER) measurements and immunocytochemical staining for platelet and endothelial cell adhesion molecule-1 and zonula occludens-1 were used to assess endothelial barrier and tight junctional integrity. LDL permeability in response to treatment with VEGF-D was measured using fluorometry in confluent HCMEC. RESULTS: Image quantitation demonstrated significant increases in VEGF-D immunoreactivity in the media of coronary arteries and intramyocardial arterioles of CAV cases, and in the intima and media of coronary arteries of DM cases. Treatment with VEGF-D, in vitro, significantly increased LDL passage through HCMEC monolayers. In conjunction, treatment with VEGF-D significantly decreased TER measurements 2 hours post treatment and induced the formation of intercellular gaps through an extracellular signal-regulated kinase 1/2 (ERK1/2)-dependent pathway. CONCLUSIONS: VEGF-D is overexpressed in the arteries of CAV and DM cases. Treatment with VEGF-D can disrupt HCMEC tight junctions, resulting in the formation of intercellular gaps, and can also significantly increase LDL permeability through confluent monolayers. PMID- 21620739 TI - Hippocampal sclerosis and encephalomalacia as prognostic factors of tuberculous meningitis-related and herpes simplex encephalitis-related epilepsy. AB - BACKGROUND: Tuberculous meningitis (TBM) and herpes simplex encephalitis (HSE) are common neurological diseases involving the brain parenchyma, and both can result in chronic epilepsy. Here, we identified possible variables affecting the prognosis of central nervous system (CNS) infection-related epilepsy. METHODS: The clinical seizure characteristics and demographic data of 20 TBM- and 55 HSE related epilepsy patients were compared. Statistically significant prognostic variables were identified using multiple regression analysis. RESULTS: Sex, age at infection, age at epilepsy onset, presence of seizures at the time of infection, latency period, and seizure characteristics between two groups were similar except for the pattern of brain lesions observed on the MRI and their overall prognosis. Patients with hippocampal sclerosis (HS) only comprised 30% and 52.7% of the TBM and HSE groups, respectively. Encephalomalacia had a positive effect in the HSE group while HS had a negative effect in this group, but no significant effects were found in the TBM group. Through a multiple regression analysis with a correction for group effects, HS was associated with a poor prognosis. However, encephalomalacia was concomitantly associated with a good prognosis. In addition, a short latency period, with a one-year interval, and being male were both associated with a good prognosis, while the age at the onset of epilepsy was associated with a poor prognosis. CONCLUSIONS: This study suggests that HS and encephalomalacia could have mutual but contradictory effects on the prognosis of CNS infection-related epilepsy. PMID- 21620740 TI - Structure of delay in carotid surgery--an observational study. AB - OBJECTIVES AND DESIGN: Undelayed investigation and surgical treatment of symptomatic carotid artery stenosis are recommended as per guidelines on stroke prevention. We evaluated patient referral pathways and delays from symptom to surgery in Helsinki University Central Hospital (HUCH) region. MATERIALS AND METHODS: One hundred consecutive symptomatic patients scheduled for carotid endarterectomy (CEA) between August 2007 and September 2008 were identified and the delay between ischaemic index symptom and CEA was analysed. RESULTS: The median time from the index symptom to surgery was 47 days (range: 3-688 days). The longest delay was surgery related with a median of 25 days (range: 2-202 days) from the consultation of the vascular surgeon to the operation. Only 11% of the patients were operated within the recommended 2 weeks' time. It was more likely that CEA was performed within 2 weeks if an emergent consultation to Meilahti Hospital neurologist on call did take place (odds ratio (OR) 12.6, 95% confidence interval (CI) 1.5-104, p = 0.019). CONCLUSION: Delays from symptom to surgery were generally too long and the in-hospital door-to-knife time (DKT) was long mostly due to waiting for the operation theatre. The investigation of all stroke, amaurosis fugax and transient ischaemic attack patients should be performed on an emergency basis and most optimally centralised to hospitals were carotid surgery is performed. PMID- 21620741 TI - Anovulatory bleeding in a 12-year-old as presenting sign for rare undifferentiated sarcoma. AB - BACKGROUND: Abnormal uterine bleeding is a common phenomenon in perimenarchal females. Though most cases of abnormal bleeding are due to anovulatory bleeding or bleeding disorders, rare cases are attributable to underlying malignancy. CASE: Here we report a 12-year-old female patient who presented with abnormal uterine bleeding three months after menarche. She was ultimately diagnosed with a pelvic sarcoma of unknown primary origin. She was subsequently treated with chemotherapy, whole pelvic radiation, and hysterectomy with bilateral salpingo oophorectomy. SUMMARY: Abnormal uterine bleeding unresponsive to conventional temporizing measures should be thoroughly evaluated. Management of pelvic malignancies in the pediatric population requires cooperation amongst specialists from an array of disciplines. PMID- 21620742 TI - Is young maternal age really a risk factor for adverse pregnancy outcome in a canadian tertiary referral hospital? AB - OBJECTIVE: To assess the risk of adverse pregnancy outcome among teenage mothers within a large tertiary referral center in Canada. METHODS: All nulliparous singleton births in the McGill University Health Centre during 2001-2007 were retrieved using the "MOND" database. Patients were divided according to maternal age: <20 years (teenage), and between 20 and 39 years. Obstetric and neonatal complications were compared. RESULTS: 9744 nulliparous women were included; 250 (2.6%) were teenage and 9494 (97.4%) were 20-39 years old. Teenage mothers tended to deliver earlier (38.0 vs 39.2 weeks gestation, P < 0.001) and had higher rates of extreme prematurity (OR 4.5, 95% CI 2.5-8.1). Babies of teenage mothers had lower birth weights (3014 g vs 3326 g, P < 0.001), higher rates of NICU admission (OR 2.1, 95% CI 1.5-3.0), congenital anomalies (OR 1.8, 95% CI 1.2-2.6) and combined perinatal and neonatal mortality (OR 3.8, 95% CI 1.9-7.5). Logistic regression analysis showed an association between young maternal age and the risk to have at least one adverse outcome (P < 0.001). CONCLUSIONS: Even within a large tertiary referral hospital, teenage mothers carry a greater risk of adverse pregnancy outcome, mainly due to preterm births. PMID- 21620743 TI - Circadian rhythms, sleep, and substance abuse. AB - Substance abuse is linked to numerous mental and physical health problems, including disturbed sleep. The association between substance use and sleep appears to be bidirectional, in that substance use may directly cause sleep disturbances, and difficulty sleeping may be a risk factor for relapse to substance use. Growing evidence similarly links substance use to disturbances in circadian rhythms, although many gaps in knowledge persist, particularly regarding whether circadian disturbance leads to substance abuse or dependence. Given the integral role circadian rhythms play in regulating sleep, circadian mechanisms may account in part for sleep-substance abuse interactions. Furthermore, a burgeoning research base supports a role for the circadian system in regulating reward processing, indicating that circadian mechanisms may be directly linked to substance abuse independently of sleep pathways. More work in this area is needed, particularly in elucidating how sleep and circadian disturbance may contribute to initiation of, and/or relapse to, substance use. Sleep and circadian-based interventions could play a critical role in the prevention and treatment of substance use disorders. PMID- 21620744 TI - Understanding the problem of poor CPAP adherence. PMID- 21620745 TI - Sleep disturbances in children with multiple disabilities. AB - INTRODUCTION: Although sleep disturbances in disabled children are of central clinical importance, there is little research on that topic. There are no data available on frequency, severity or aetiology of sleep disturbances and related symptoms in this specific patient group. OBJECTIVE: To review the current state of research and outline future research objectives. METHODS: We searched international scientific databases for relevant publications from 1980-2009. From all papers qualifying for further analysis we retrieved systematic information on sample characteristics, sleep assessment tools and their test quality criteria, and core findings. RESULTS: 61 publications including 4392 patients were categorized as "mixed" (reporting on heterogeneous diagnoses), or "specified" papers (specific diagnoses) based on international classification of diseases (ICD) 10 classification. To assess sleep disturbances, most authors relied on subjective instruments with poor psychometric quality. Mean prevalence of sleep disturbances was 67% (76%,"mixed" group; 65%, "specified" group). In children suffering severe global cerebral injury, the prevalence of sleep disturbances was even higher (>90%). The most frequent symptoms were insomnia and sleep-related respiratory disorders. Some of these symptoms were closely associated with specific medical syndromes. CONCLUSION: There is an urgent need for sleep disturbance assessment tools evaluated for the patient group of interest. By use of validated assessment tools, patient factors, which may be crucial in causing sleep disturbances, may be investigated and appropriate treatment strategies may be developed. PMID- 21620746 TI - Is evidence-based medicine so evident in veterinary research and practice? History, obstacles and perspectives. AB - Evidence-based medicine (EBM) refers to the conscientious, explicit and judicious use of current best evidence from research for the care of an individual patient. The concept of EBM was first described in human medicine in the early 1990s and was introduced to veterinary medicine 10 years later. However, it is not clear that the EBM approach promulgated in human medicine can be applied to the same extent to veterinary medicine. EBM has the potential to help veterinarians to make more informed decisions, but obstacles to the implementation of EBM include a lack of high quality patient-centred research, the need for basic understanding of clinical epidemiology by veterinarians, the absence of adequate searching techniques and accessibility to scientific data bases and the inadequacy of EBM tools that can be applied to the busy daily practise of veterinarians. This review describes the development of EBM in the veterinary profession, identifies its advantages and disadvantages and discusses whether and how veterinary surgeons should further adopt the EBM approach of human medicine. PMID- 21620747 TI - The relationship between self-reported severe pain and measures of socio-economic disadvantage. AB - AIMS: To determine the association of severe pain with socioeconomic characteristics. METHODS: Data was extracted from the Health Survey for England (HSE), 2005. The HSE is a series of annual cross-sectional surveys designed to describe the health of people living in private homes in England, from a random sample of 720 postcode sectors. Interviewees were >= 16 years. Pain severity was characterised by the EQ-5D. Socioeconomic status was classified by ability to work, social security benefits, the National Statistics Socio-economic Classification (NS-SEC) and the Index of Multiple Deprivation (IMD). RESULTS: Pain status was recorded for 9419 subjects of whom 431 (4.6%) reported severe pain. 156 of those reporting severe pain were of working age. Of these 68 (43.6%) stated they were unable to work due to sickness or disability and 64 (41.0%) claimed a state benefit. After adjusting for disease and demographic variables, severe pain was associated with the IMD with an odds ratio of 1.65 (95% CI 1.16 2.34, p=0.005), NS-SEC (OR=2.94; 95% CI 1.76-4.91) and equivalised household income (lowest versus highest quintile; OR=2.58 (95% CI 1.46-4.57, p=0.001). CONCLUSIONS: This study demonstrated significant associations between pain and socio-economic disadvantage. Apart from the direct impact upon the individual, this clearly has wider societal implications in terms of additional health and social care costs for affected people. PMID- 21620748 TI - A 3-year randomized therapeutic trial of nitisinone in alkaptonuria. AB - Alkaptonuria is a rare, autosomal recessive disorder of tyrosine degradation due to deficiency of the third enzyme in the catabolic pathway. As a result, homogentisic acid (HGA) accumulates and is excreted in gram quantities in the urine, which turns dark upon alkalization. The first symptoms, occurring in early adulthood, involve a painful, progressively debilitating arthritis of the spine and large joints. Cardiac valvular disease and renal and prostate stones occur later. Previously suggested therapies have failed to show benefit, and management remains symptomatic. Nitisinone, a potent inhibitor of the second enzyme in the tyrosine catabolic pathway, is considered a potential therapy; proof-of-principle studies showed 95% reduction in urinary HGA. Based on those findings, a prospective, randomized clinical trial was initiated in 2005 to evaluate 40 patients over a 36-month period. The primary outcome parameter was hip total range of motion with measures of musculoskeletal function serving as secondary parameters. Biochemically, this study consistently demonstrated 95% reduction of HGA in urine and plasma over the course of 3 years. Clinically, primary and secondary parameters did not prove benefit from the medication. Side effects were infrequent. This trial illustrates the remarkable tolerability of nitisinone, its biochemical efficacy, and the need to investigate its use in younger individuals prior to development of debilitating arthritis. PMID- 21620749 TI - CNS-targeted AAV5 gene transfer results in global dispersal of vector and prevention of morphological and function deterioration in CNS of globoid cell leukodystrophy mouse model. AB - Globoid cell leukodystrophy (GLD) is a devastating lysosomal storage disease caused by deficiency of the enzyme galactocerebrosidase (GALC). Currently, there is no definite cure for GLD. Several attempts with CNS-directed gene therapy in twitcher mice (a murine model of GLD) demonstrated restricted expression of GALC activity in CNS and failure of therapeutic efficacy in cerebellum and spinal cord, resulting in various degrees of correction of biochemical, pathological and clinical phenotype. More recently, twitcher mice receiving a combination of hematopoietic and viral vector gene transfer therapies were not protected from neurodegeneration and axonopathy in both cerebellum and spinal cord. This evidence indicates the requirement of sufficient and widespread GALC expression in CNS and rescue of cerebellum and spinal cord in the therapeutic intervention of murine model of GLD. In this study, we have optimized intracranial delivery of AAV2/5-GALC to the neocortex, hippocampus and cerebellum, instead of the thalamus as was previously conducted, of twitcher mice. The CNS-targeted AAV2/5 gene transfer effectively dispersed GALC transgene along the neuraxis of CNS as far as the lumbar spinal cord, and reduced the accumulation of psychosine in the CNS of twitcher mice. Most importantly, the treated twitcher mice were protected from loss of oligodendrocytes and Purkinje cells, axonopathy and marked gliosis, and had significantly improved neuromotor function and prolonged lifespan. These preclinical findings with our approach are encouraging, although a more robust response in the spinal cord would be desirable. Collectively, the information in this study validates the efficacy of this gene delivery approach to correct enzymatic deficiency, psychosine accumulation and neuropathy in CNS of GLD. Combining cell therapy such as bone marrow transplantation with treatment with the aim of reducing inflammation, replacing dead or dying oligodendrocytes and targeting PNS may provide a synergistic and more complete correction of this disease. PMID- 21620750 TI - Sam68 is required for both NF-kappaB activation and apoptosis signaling by the TNF receptor. AB - The RNA-binding protein Sam68 is implicated in various cellular processes including RNA metabolism, apoptosis, and signal transduction. Here we identify a role of Sam68 in TNF-induced NF-kappaB activation and apoptosis. We found that Sam68 is recruited to the TNF receptor, and its deficiency dramatically reduces RIP recruitment and ubiquitylation. It also impairs cIAP1 recruitment and maintenance of recruited TRAF2 at the TNF receptor. In its absence, activation of the TAK1-IKK kinase complex is defective, greatly reducing signal transduction. Sam68 is also found as a part of the TNF-induced cytoplasmic caspase-8-FADD complex. RIP is not recruited to this complex in Sam68 knockout cells, and caspase activation is virtually absent. These findings delineate previously unknown functions for Sam68 in the TNF signaling pathway, where it acts as a signaling adaptor both in the membrane-associated complex I and in the cytoplasmic complex II, regulating both NF-kappaB activation and apoptosis. PMID- 21620751 TI - [Assessment of maternal satisfaction with epidural analgesia during labour]. AB - OBJECTIVE: To measure the overall satisfaction of obstetrics patients with epidural analgesia during labour, and in particular, with the technique and other subjective factors. MATERIAL AND METHODS: An anonymous questionnaire was administered over a three-month period to patients who received analgesia to control pain during labour and who had vaginal delivery, in order to obtain information of the satisfaction with technique and their care. RESULTS: Of the 100 patients who responded to the questionnaire, 92% were satisfied with the technique. The mild satisfaction measured by SERVQHOS score was 3.98 (SD +/- 0.64). The score for the subjective aspects was 4.10 (+/- 0.68), which was better than objective ones. The recommendation rate was 98% for satisfied patients and 85.7% for those who were not satisfied. There were no differences between Spanish and foreign patients in their evaluation of the satisfaction. CONCLUSIONS: The proportion of patients satisfied with the technique was very high, and was the top rated subjective aspect (treatment and public confidence). PMID- 21620752 TI - [Analytic study of hopes and perceptions of second-year medical school students during gross anatomy laboratory cessions]. AB - INTRODUCTION: Using an anonymous questionnaire, this study aimed to assess hopes and perceptions of second-year medical school students faced to gross anatomy dissections and to appreciate the status of dissection within the others anatomical teaching tools. MATERIALS AND METHODS: The questionnaire was distributed among 210 second-year medical students during gross anatomy dissections. The first part of the questionnaire aimed to assess perceptions of students before dissections while the second part aimed to position dissection in their anatomical teaching cursus. RESULTS: Within the 210 students participating to the study, 70 (45%) had not seen a dead human body before the laboratory cession. The main stressful factors were technical difficulties encountered during dissection more than embarrassment in front of the cadaver. The level of stress was significantly higher in female students. Hand, neck and gluteal regions were the most stressful regions reported by students. Students considered that gross anatomy dissection was a key ritual experience crucial for their formation that should not be only proposed to senior practitioners. CONCLUSION: This study illustrates the invariable interest of medical students for gross anatomy laboratory cessions that permit a first technical experience, faced to the death. PMID- 21620753 TI - Bone metastasis: histological changes and pathophysiological mechanisms in osteolytic or osteosclerotic localizations. A review. AB - The development of a bone metastasis involves interactions between the tumor cells, the bone marrow microenvironment and the bone cells themselves. A better understanding of the pathophysiological changes occurring in bone metastasis can be obtained from histopathological examination of invaded specimens. This review focuses on the main molecular mechanisms implied in the localization and growth of malignant cells in the bone marrow. The corresponding histologic developmental stages are illustrated both in osteolytic (or mixed metastasis) or in the osteosclerotic forms by histological analysis, immunohistochemistry and microcomputed tomographic analysis of bone samples. In both cases, the malignant cells find a "fertile soil" in the bone marrow microenvironment. They use the growth factors released by bone cells for the coupling between osteoclasts/osteoblasts to promote their own development. In turn, they elaborate a variety of cytokines that can promote osteoclastogenesis (PTHrP, IL-1, IL-6...) or on the contrary, other growth factors that can boost the osteoblastic activity (ET1, IGFs). A "vicious circle" occurs between the malignant cells and the bone cells leading to the radiological expression of the metastasis. PMID- 21620754 TI - Does milling one-piece titanium dental implants induce osteocyte and osteoclast changes? AB - One-piece dental implants avoid adverse effects sometimes associated with the traditional implant-abutment interface and may provide a suitable alternative to two-piece implants; however, one-piece implants often need in situ milling, which may exacerbate cell apoptosis from excessive heat at the bone-implant interface and induce secondary crestal bone loss. Twelve implants were placed in the metaphyses of two sheep under general anesthesia. Six implants were milled with a diamond bur while the other six implants remained intact. Animals were euthanized after four days, and bone blocks were harvested. Bone samples were studied without decalcification. Osteocytes were stained with Hoechst 33342 and osteoclasts by the TRAcP reaction. Both cell types, in the cortical and trabecular bone around the implant's cervical region, were counted utilizing morphometric methods. Values were compared to areas at a distance from the cervical region. No difference was observed between milled and unmilled implants, which suggested that the amount of generated heat did not provoke osteocyte loss or induce osteoclastogenesis. Intraoral abutment preparations did not increase cellular apoptosis at the bone-implant interface after four days in the ovine model. PMID- 21620755 TI - [Arthroscopic assessment of articular fractures of distal radius osteosyntheses by percutaneous pins]. AB - OBJECTIVES: We studied the radiological results of percutaneous pinning in articular fractures of the distal radius. We did not, thereby, have a method of direct ligamentous and articular assessment. We realized an anatomical arthroscopic assessment at six weeks of osteosynthesis of wrists. The purpose was to study the correlation between the presence of anatomical lesions and the functional result. METHODS: Thirty-five patients presenting an articular fracture with posterior displacement were included; 30 women and five men. The mean age was 62 years. All the patients were right -handed and this was the fractured side in 18 patients. The patients had clinical and radiological follow-up at three, six, 12 weeks and one year. Surgical techniques were Py or Kapandji pinning. An arthroscopy of the wrist was carried out at six weeks from the fixation at pin removal. An assessment of ligamentous lesions and quality of articular reduction was performed. RESULTS: Twenty-eight per cent of the patients had ligamentous lesions. Nine patients presented an interfragmentary distance greater than 1mm. Two patients had an articular step as high as 2mm. CONCLUSIONS: Clinical examination did not show any correlation between the anatomical imperfections noted at the time of arthroscopy and the function of the wrist at one year postoperative, regardless of the technique used. PMID- 21620756 TI - Effects of monopolar and bipolar electrode configurations on surface EMG spike analysis. AB - This study compared the effects of monopolar and bipolar electrode configurations on interference pattern analysis of the surface electromyographic (sEMG). Twenty four college-aged male participants performed isometric actions of the elbow flexors at 40, 60, 80, and 100 percent of maximal voluntary contraction (MVC). Separate (Ag/AgCl) electrodes were used for both configurations. There were five measures associated with "spike shape" analysis: mean spike amplitude (MSA), mean spike frequency (MSF), mean spike slope (MSS), mean spike duration (MSD) and mean number of peaks per spike (MNPPS). A load-cell and wrist-cuff assembly was used to record isometric elbow flexion forces. Both electrode configurations resulted in the same trends force changes in spike shape measures across force levels: there was a linear increase in MSA, MSS, and a quadratic decrease in MSF and the MNPPS (p's<0.05). The MSD underwent a quadratic increase (p<0.05). The spike shape measures had greater mean magnitudes and exhibited greater rates of changes across force levels for the monopolar electrode configuration (p's<0.05). The monopolar electrode configuration was therefore more sensitive to changes in muscle activity with increases in isometric force. This is an important consideration because the rate at which muscle electrical activity develops into a full interference pattern is an important qualitative and quantitative diagnostic measure. PMID- 21620757 TI - Painful legs and moving toes syndrome associated with a sacral Tarlov cyst. PMID- 21620758 TI - To what extent can deprivation inequalities in mortality and heart disease incidence amongst the working aged in Scotland be explained by smoking? Relative and absolute approaches. AB - Smoking is important for both population health and inequalities in health. There is a growing recognition that its impact on inequalities can be assessed both by standardising smoking rates across socio-economic groups and by assessing the effect of reducing the prevalence of smoking in all groups, so-called relative and absolute approaches. While national level studies (such as census-linkage studies) give vital information on the socio-economic gradient in health they often lack smoking data. Here, small area smoking estimates are linked to a national level longitudinal study to overcome this problem. Results confirm that in relative and especially absolute terms smoking plays an important role in inequalities. PMID- 21620759 TI - Effect of the resonance of the C-H and O-H bond stretching vibrations on the IR spectra of the hydrogen bond in formic and acetic acid. AB - It is shown that the resonance of the O-H and C-H bond stretching vibrations is responsible for a noticeable intensity redistribution effect in the IR spectra of associated formic acid molecules in the gaseous phase. This effect is manifested by a considerably high growth in intensity of the nuC-H band, which overlaps the nuO-H band contour in the spectra. A vibronic coupling of the Herzberg-Teller type expressed by the second order term in the perturbation theory is the most probable source of these spectral effects. The presented mechanism explains the variation of the effect magnitude accompanying the phase transitions. The proposed model also facilitates the understanding of the H/D isotopic effects in the spectra as well as the essential difference in the corresponding spectral properties between the formic and the acetic acid. PMID- 21620760 TI - Theoretical study of electronic structure and absorption spectra of diacid and zinc species of series of meso-phenylporphyrins. AB - DFT and TDDFT calculations with density functionals (PBE1PBE, B3LYP, and PBEPBE) have been employed in a study of HCl-acidified diacids of a series of meso phenylporphyrins. This study aims to clarify the influence of conformational distortion, meso-phenyl substituents, and counterion Cl- on absorption spectra of porphyrin derivatives. Calculations indicate that all three factors increase the MO's level and decrease the Gouterman HOMO-LUMO gap; this, further, brings about the redshift of absorption band. In comparison with experimental methods, the PBE1PBE method produces a more credible description of UV-vis spectra than other two methods. TDDFT calculation with the PBE1PBE method indicates that the electronic effect of the meso-phenyl group is dominant for the spectral redshift of porphyrin diacid series as observed in zinc porphyrins. The redshift of the B band of porphyrin diacids is primarily caused by an electron-donating effect of the meso-phenyl group, whereas the Q band is more sensitive to the pi electron delocalization effect. The counterion is indispensable in a theoretical study of electronic and spectral structure of porphyrin diacids. PMID- 21620762 TI - Conformational preferences for some 5-substituted 2-acetylthiophenes through infrared spectroscopy and theoretical calculations. AB - The s-cis-trans isomerisms of some derivatives of thiophene (2-acetyl, AT; 2 acetyl-5-bromo, ABT and 2-acetyl-5-chloro, ACT) were analyzed, using data from deconvolution of their carbonyl absorption bands in two solvents (CCl4 and CHCl3). These infrared data showed that the O,S-cis conformer largely predominates in the studied solvents and that the same occurs in the gas phase, as observed from theoretical calculations. The latter results were obtained using B3LYP/6-311++G(3df,3p) and MP2/6-311++G(3df,3p) levels of theory, with zero-point energy correction. Moreover, the use of the IEFPCM (Integral Equation Formalism Polarizable Continuum Model) to take into account the solvent effects, using the same levels of theory, confirmed the results observed from infrared data. Low temperature 13C NMR spectra in CS2/CD2Cl2 (-90 degrees C) and in acetone-d6 (-80 degrees C) did not show pairs of signals for each carbon, due to the known low energy barrier (~8 kcal mol(-1)) for the cis-trans interconversion. Data from NBO calculations show that the nO(2)->sigmaS-C5* and nO(2)->sigmaC2-C3* interactions occur only in the O,S-cis isomer and can explain its conformational preference. PMID- 21620761 TI - Oxidation-extraction spectrometry of reactive oxygen species (ROS) generated by chlorophyllin magnesium (Chl-Mg) under ultrasonic irradiation. AB - In order to examine the mechanism and process of sonodynamic reaction, the chlorophyllin magnesium (Chl-Mg) acting as a sonosensitizer was irradiated by ultrasound, and the generation of reactive oxygen species (ROS) were detected by the method of oxidation-extraction spectrometry (OES). That is, under ultrasonic irradiation in the presence of Chl-Mg, the 1,5-diphenyl carbazide (DPCI) is oxidized by generated ROS into 1,5-diphenyl carbazone (DPCO), which can be extracted by mixed organic solvent and display a obvious visible absorption at 563 nm wavelength. Besides, the generation conditions of ROS were also reviewed. The results demonstrated that the quantities of generated ROS increased with the increase of ultrasonic irradiation time, Chl-Mg concentration and DPCI concentration. Finally, several radical scavengers (l-Histidine (His), 2,6-Di tert-butyl-methylphenol (BHT) and Vitamin C (VC)) were used to determine the kind of the generated ROS. It was found that at least the hydroxyl radical (OH) and singlet oxygen (1O2) were generated in the presence of Chl-Mg under ultrasonic irradiation. It is wish that this paper might offer some valuable references for the study on the mechanism of SDT and the application of Chl-Mg in tumor treatment. PMID- 21620763 TI - Synthesis, vibrational spectral and nonlinear optical studies of N-(4-hydroxy phenyl)-2-hydroxybenzaldehyde-imine: a combined experimental and theoretical investigation. AB - The study of imine-bridged organics has been the one hot spot of photo-responsive material sciences in recent years. Herein we make a study of the synthesis, characteristics and potential application of N-(4-hydroxy-phenyl)-2-hydroxy benzaldehyde-imine (HPHBI), C13H11NO2. The studied compound was synthesized in one step by the condensation reaction of salicylaldehyde and 4-aminophenol in methanol solution, and characterized by single crystal X-ray diffraction, FT-IR and FT-Raman techniques with theoretical calculations at B3LYP/6-31G(d) level. The molecule adopts trans configuration about central CN bond with intramolecular hydrogen bonding, and the adjacent molecules form wave-shaped structure linked by strong intermolecular hydrogen bonding mechanism along b axis. The vibrational spectra have been precisely assigned with the aid of theoretical frequencies. Furthermore, the thermodynamic properties have been obtained by the theoretical vibrational analysis for HPHBI. The total linear polarizability and first-order hyperpolarizabilities calculated on the studied compound respectively present 25.378 A3 and 1.655*10(-29) cm5/esu, which indicates the compound has relatively good nonlinear optical property. PMID- 21620764 TI - Cytomegalovirus quantification: where to next in optimising patient management? AB - BACKGROUND: Over the years quantification of cytomegalovirus (HCMV) load in blood has become a mainstay of clinical management helping direct deployment of antiviral therapy, assess response to therapy and highlight cases of drug resistance. AIMS: The review focuses on a brief historical perspective of HCMV quantification and the ways in which viral load is being used to improve patient management. METHODS: A review of the published literature and also personal experience at the Royal Free Hospital. RESULTS: Quantification of HCMV is essential for efficient patient management. The ability to use real time quantitative PCR to drive pre-emptive therapy has improved patient management after transplantation although the threshold viral loads for deployment differ between laboratories. The field would benefit from access to a universal standard for quantification. CONCLUSIONS: We see that HCMV quantification will continue to be central to delivering individualised patient management and facilitating multicentre trials of new antiviral agents and vaccines in a variety of clinical settings. PMID- 21620765 TI - Reduced mismatch negativity and increased variability of brain activity in schizophrenia. AB - OBJECTIVES: Schizophrenia patients commonly exhibit smaller amplitudes of mismatch negativity (MMN) than in controls. It remains unclear whether this results from deficient processes indexed by MMN or 'normally' though more variable processing. The present magnetoencephalographic study addressed this question by analyzing intra-individual trial-by-trial variability and MMN amplitude. METHODS: Twenty inpatients meeting ICD criteria for schizophrenia and 18 healthy controls participated in an auditory oddball experiment. The neuromagnetic mismatch field (MMNm) was defined as the difference waveform deviant minus standard tone response. Variability index (VI) in different frequency bands was quantified as trial-by-trial variation of stimulus-evoked responses and epoch-by-epoch variation of signal amplitude during a resting condition. RESULTS: Patients displayed a smaller MMNm amplitude and higher VI during the oddball experiment and during the resting condition than in controls. VI and MMNm amplitude were correlated in controls, but not in patients. CONCLUSION: Reduced MMN in schizophrenia cannot be explained by augmented variability of brain activity; deficient auditory sensory memory and stimulus related phase-locking may characterize the disorder. SIGNIFICANCE: Understanding the contribution of diminished temporal stability of neuronal network dynamics to schizophrenia is crucial in modeling the impact of such instability on performance and thus for understanding deviant attention and memory functions. PMID- 21620767 TI - Physical fitness in rural and urban children and adolescents from Spain. AB - OBJECTIVE: We aimed to examine the differences in fitness components between rural and urban Spanish children and adolescents. DESIGN/METHOD: A sample of 2569 school children (1068) and adolescents (1501) from urban and rural areas from the region of Aragon (Spain). Physical fitness was assessed by 7 tests: 20 m shuttle run, speed shuttle run, sit and reach, standing long jump, handgrip strength, bent arm hang and sit-ups in 30s (s). Differences (in fitness and anthropometry) by place of residence were examined by one-way analysis of covariance. RESULTS: Rural young people had higher cardiorespiratory fitness, but lower speed-agility and flexibility than urban young people (all p<0.001; mean differences=1.5 mL/kg per min, 0.4s and 0.9 cm respectively). Regarding muscular fitness, rural young people had a better performance in handgrip strength (mean difference=0.8 kg), bent arm hang (mean difference=2.3s), and a lower performance in sit-ups in 30s (mean difference=0.9s) (all p<0.001), compared to their urban peers. Rural young people had a lower body mass, body mass index (BMI) and sum of skinfolds (all p<0.05; mean differences=1.1 kg, 0.3 kg/m(2) and 4.9 mm respectively) than their urban peers. Effect size was small-medium (Cohen's d=0.1-0.3). CONCLUSION: Rural Spanish children and adolescents had overall a healthier profile than their urban peers in terms of cardiorespiratory fitness, upper- and lower-limb muscular fitness and adiposity, while they performed worse in speed-agility and flexibility. The place of residence should be taken into account when implementing effective interventions to promote physical activity and health. PMID- 21620766 TI - Aggressive behavior, bullying, snoring, and sleepiness in schoolchildren. AB - BACKGROUND: To assess whether urban schoolchildren with aggressive behavior are more likely than peers to have symptoms suggestive of sleep-disordered breathing. METHODS: Cross-sectional survey of sleep and behavior in schoolchildren. Validated screening assessments for conduct problems (Connor's rating scale), bullying behavior, and sleep-disordered breathing (pediatric sleep questionnaire) were completed by parents. Teachers completed Connor's teacher rating scale. RESULTS: Among 341 subjects (51% female), 110 (32%) were rated by a parent or teacher as having a conduct problem (T-score ?65) and 78 (23%) had symptoms suggestive of sleep-disordered breathing. Children with conduct problems, bullying, or discipline referrals, in comparison to non-aggressive peers, more often had symptoms suggestive of sleep-disordered breathing (each p<0.05). Children with vs. without conduct problems were more likely to snore habitually (p<0.5). However, a sleepiness subscale alone, and not a snoring subscale, predicted conduct problems after accounting for age, gender, a measure of socioeconomic status, and stimulant use. CONCLUSIONS: Urban schoolchildren with aggressive behaviors may have symptoms of sleep-disordered breathing with disproportionate frequency. Sleepiness may impair emotional regulation necessary to control aggression. PMID- 21620768 TI - Regulatory T cells: stability revisited. AB - Breakdown in self-tolerance is caused, in part, by loss of regulatory T (Treg) cells. Recently, a controversy has surfaced about whether Treg cells are overwhelmingly stable, or if they can be reprogrammed in inflammatory and autoimmune environments. Those in the instability camp have shown that a fraction of Treg cells lose forkhead box P3 protein and acquire effector arm activities. Instability is coupled with interleukin-2 insufficiency and the inflammatory milieu that promotes reprogramming. Here, we highlight the basic tenets of each viewpoint and discuss technical, biological and environmental differences in the models that might help yield a unifying hypothesis. Also considered is how Treg cell instability could link to development of autoimmune disease and the implications for trials of Treg cell-based therapy. PMID- 21620769 TI - ER stress in retinal degeneration: a target for rational therapy? AB - Mutations that cause rhodopsin misfolding and retention within the endoplasmic reticulum (ER) are a prominent cause of retinitis pigmentosa. Here, we discuss the hypothesis that the failure of photoreceptor neurons to adapt to the stress caused by rhodopsin accumulation in the ER leads to a global collapse of homeostasis and to retinal degeneration. We review the molecular mechanisms underlying the activity of local ER conformational sensors and stress-relaying modules and consider how ER-derived stress signals are amplified and implemented to impact on downstream processes, including rhodopsin clearance and cell fate control. The emerging view is that alterations to the systems responsible for the detection, transduction and implementation of ER stress might be used therapeutically to treat retinitis pigmentosa. PMID- 21620770 TI - Everyday verbal memory and pediatric epilepsy. AB - This study addressed the reliability and validity of reports of everyday verbal memory with a sample of 132 pediatric patients with epilepsy. Each patient and one parent completed a questionnaire on everyday verbal memory comprising two scales assessing learning/retrieval and prospective memory. Each patient was also administered tests of memory, attention, and academic skills. Information about attention, mood, and academic performance was obtained from parent and teacher report, as well as self-report. Memory test scores were correlated with children's reports of learning and retrieval in everyday activities, but were not significantly associated with reports of prospective memory. Reports of everyday memory were found to be reliable and predictive of academic performance. Performance on tests of memory, conversely, was unrelated to reports of academic performance. Reports of everyday memory may, therefore, provide more useful information than tests when evaluating the effects of epilepsy and its treatments. PMID- 21620771 TI - Low long-term efficacy and tolerability of add-on rufinamide in patients with Dravet syndrome. AB - In this retrospective European multicenter study we evaluated the efficacy and tolerability of rufinamide in patients with Dravet syndrome and refractory seizures. Twenty patients were included; in 16 patients a SCN1A mutation was detected. The responder rate after 6 months was 20%, and after 34 months, 5%. The retention rate was 45% after 6 months and 5% after 34 months. Rufinamide treatment was stopped because of aggravation of seizures (30%), no effect (45%), and side effects (10%). The efficacy and long-term retention rate were low in our patients with Dravet syndrome and refractory seizures, far lower than in patients with Lennox-Gastaut syndrome; one-third of our patients experienced seizure aggravation. Therefore, rufinamide does not seem to be a suitable option for long term treatment in patients with Dravet syndrome. PMID- 21620772 TI - Suicidality and epilepsy: a neuropsychobiological perspective. AB - People with epilepsy (PWE) are at increased risk of experiencing suicidal ideation, displaying suicidal behavior, and committing suicide than the general population. The relationship between suicidality and epilepsy is complex and multifactorial in which operant pathogenic mechanisms include epilepsy-related variables, personal and familial psychiatric history, and iatrogenic effects. Furthermore, a bidirectional relationship between suicidality and epilepsy has suggested the existence of common neurobiological pathogenic mechanisms operant in both conditions and including disturbances of several neurotransmitters, in particular, serotonin (5HT), norepinephrine (NE), glutamate (GTE), and gamma aminobutyric acid (GABA), and disturbances of the hypothalamic-pituitary-adrenal axis (HPAA), which, in turn, can result in abnormal secretion of some of these neurotransmitters. The purpose of this article is to review these common neurobiological pathogenic mechanisms. PMID- 21620773 TI - Autism in Dravet syndrome: prevalence, features, and relationship to the clinical characteristics of epilepsy and mental retardation. AB - Autism is a pervasive developmental disorder that frequently co-occurs with epilepsy. Dravet syndrome is a severe epileptic encephalopathy associated with psychomotor developmental delay. Autism in Dravet syndrome, however, has rarely been studied. In this study, the prevalence and features of autism in patients with Dravet syndrome, their potential association with mental retardation, and the clinical characteristics of epilepsy were investigated. Clinical data of 37 patients with Dravet syndrome were collected, and evaluations of autism and mental retardation were performed. Nine patients (24.3%) met the criteria for autism. All patients with autism showed speech delay, no emotional reciprocity, and narrow interests, whereas 89.3, 46.4, and 39.9% of patients without autism had speech delay, short temper, and narrow interests, respectively. Mental retardation was observed in 94.6% of patients with Dravet syndrome, with more frequent severe or profound mental retardation in those with autism. The clinical features of epilepsy did not statistically differ between the patients with autism and the patients without autism. PMID- 21620774 TI - Extralimbic autoimmune encephalitis associated with glutamic acid decarboxylase antibodies: an underdiagnosed entity? AB - Nonparaneoplastic glutamic acid decarboxylase antibody (GADAb)-related autoimmune encephalitis is a syndrome characterized by refractory seizures, progressive cognitive deficits, and psychiatric manifestations. The limbic subtype is well described, has characteristic affective and memory disturbances, and typical mesial temporal MRI abnormalities. We found only one single case report of the extralimbic subtype. We report clinical, radiological, and pathological findings of two additional cases with contrast-enhancing lesions. One of our cases presented as vasculitis, and the other imitated a tumor. Pathological evidence of both vasculitis and encephalitis has never been previously reported in any inflammatory condition affecting the brain. Our cases confirm prior reports that immune therapy can better control seizures associated with GADAb autoimmune encephalitis, and support the rationale for assaying for GADAb titers in patients with etiologically unclear extralimbic lesions and refractory epilepsy, independent of seizure types. PMID- 21620775 TI - Socio-occupational and employment profile of patients with epilepsy. AB - OBJECTIVE: Epilepsy has a significant impact on quality of life. Many studies have observed higher unemployment rates among patients with epilepsy. However, unemployment rates vary according to the clinical conditions, country, and group studied. METHODS: Between October 2007 and February 2008, we performed a cross sectional multicenter epidemiological study to evaluate the socio-occupational and employment profiles of 872 adult patients with epilepsy followed in outpatient epilepsy clinics in Spain. RESULTS: Fifty-eight percent of the patients were employed at the time of the survey, 10.9% of the patients were unemployed, and 12.5% were occupationally incapacitated. CONCLUSION: Patients with epilepsy had employment rates similar to those of the general population, and slightly higher levels of unemployment. The main factors associated with unemployment and incapacity were the presence of refractory epilepsy, the occurrence of a seizure in the last 12 months, level of education, and polytherapy. PMID- 21620776 TI - Fusion rates of a morselized local bone graft in polyetheretherketone cages in posterior lumbar interbody fusion by quantitative analysis using consecutive three-dimensional computed tomography scans. AB - BACKGROUND CONTEXT: Posterior lumbar interbody fusion (PLIF) using harvested local bone inserted into a polyetheretherketone (PEEK) cage is a commonly used procedure, but the accurate fusion rate of a cage, cage to bone contact area ratio, and the changes in fusion rate with time after surgery are unknown. PURPOSE: The aim was to conduct a quantitative analysis of the fusion rates and the cage to bone contact area ratios at each period of time using a PEEK cage in PLIF using a consecutive three-dimensional (3D) computed tomography (CT) scan. STUDY DESIGN: This was a prospective study using a consecutive 3D thin-section CT scan. PATIENT SAMPLE: Thirty patients aged between 37 and 73 years set to undergo elective PLIF with PEEK cages and pedicle screw fixation were included in the study. OUTCOME MEASURES: The assessments included the Korean Oswestry Disability Index (K-ODI), Short Form (SF)-36 questionnaire, Visual Analog Scale (VAS) pain score, and dynamic plane radiographs, preoperatively and at 6 and 12 months after surgery. METHODS: Three-dimensional CT scans were performed at 6 and 12 months after surgery. Three-dimensional CT assessments of the following were performed: fusion rate of the cage, cage to bone contact area ratio, and fusion rate of the interbody bone graft besides the cage. RESULTS: The 6-month fusion rate of the segment was 86.7%, which increased to 90.0% at 12 months. The fusion area ratio between the cage area and end plate showed a significant increase from 52.0% at 6 months to 58.5% at 12 months. Regarding the fusion area ratio between the cage and end plate, the ratio between the lower surface of the cage was significantly higher than that of the upper surface. In addition, the K-ODI, SF-36, and VAS values were similar at 6 and 12 months after surgery. CONCLUSIONS: The fusion rate of the PEEK cage used in PLIF measured at 12 months was higher than that measured at 6 months. Therefore, an assessment on the complete fusion of local bone at 12 months after surgery is more accurate. PMID- 21620778 TI - Quality of life after sleeve gastrectomy and adjustable gastric banding. AB - BACKGROUND: With the addition of laparoscopic vertical sleeve gastrectomy (SG) to the bariatric surgery procedural toolkit, patients desiring a restrictive bariatric procedure often choose between adjustable gastric banding (LAGB) and SG. One study compared quality of life after these 2 procedures and found no difference. The purpose of our study was to re-evaluate the postoperative quality of life in LAGB and SG patients at a military teaching hospital in the United States. METHODS: A retrospective review of 108 consecutive laparoscopic restrictive bariatric procedures performed within 15 months at a Department of Defense hospital was conducted. Of these 108 patients, 69 had undergone laparoscopic vertical SG and 39 LAGB. A validated quality of life questionnaire (Bariatric Quality of Life) was conducted a mean of 9.3 +/- 3.2 months (range 5 16) postoperatively. The weight loss and standard laboratory parameters were measured at 0, 1, 3, 6, and 12 months. RESULTS: The quality of life assessment revealed significantly better scores after SG than after LAGB (66.5 versus 57.9, P = .0002). The excess weight loss and excess body mass index loss at 3, 6, and 12 months postoperatively were significantly greater in the laparoscopic SG group. The patients demonstrated a clear preference over time for SG once it was offered. CONCLUSION: Early postoperative quality of life was superior after SG than after LAGB. SG also resulted in superior early excess weight loss. In a practice not constrained by reimbursement, these findings were associated with increased patient choice of SG after it began to be offered. PMID- 21620777 TI - Cell autonomous role of PTEN in regulating castration-resistant prostate cancer growth. AB - Alteration of the PTEN/PI3K pathway is associated with late-stage and castrate resistant prostate cancer (CRPC). However, how PTEN loss is involved in CRPC development is not clear. Here, we show that castration-resistant growth is an intrinsic property of Pten null prostate cancer (CaP) cells, independent of cancer development stage. PTEN loss suppresses androgen-responsive gene expressions by modulating androgen receptor (AR) transcription factor activity. Conditional deletion of Ar in the epithelium promotes the proliferation of Pten null cancer cells, at least in part, by downregulating the androgen-responsive gene Fkbp5 and preventing PHLPP-mediated AKT inhibition. Our findings identify PI3K and AR pathway crosstalk as a mechanism of CRPC development, with potentially important implications for CaP etiology and therapy. PMID- 21620779 TI - Role of eutopic endometrium in pelvic endometriosis. AB - Endometriosis is defined as the presence of endometrial tissue outside the uterine cavity and is considered a benign gynecologic condition; however, in some cases, it may be aggressive. The pathogenesis of endometriosis is complex and multifactorial. Despite being one of the most widely studied gynecologic diseases, its pathogenesis remains uncertain. The intrinsic endometrial abnormalities thought to be associated with endometriosis include abnormal expression of genes, modification of endometrial response to hormones such as progesterone; increased nerve density, and oxidative stress. Evaluation of the endometrium in patients with endometriosis is an important line of investigation in the pathophysiology of the disease. It has been suggested that investigation of eutopic endometrium may help to achieve this goal. Presented herein is a literature review and a comprehensive evaluation of the role of eutopic endometrium in pelvic endometriosis. Clinical correlations of the disease are highlighted, with the objective of understanding the role of eutopic endometrium in endometriosis. PMID- 21620780 TI - Acupuncture attenuates exercise-induced increases in skin sympathetic nerve activity. AB - To identify the effect of acupuncture on skin sympathetic nerve activity (SSNA), 17 healthy subjects (7 male and 10 female) underwent LI4 acupuncture and sham needle insertion during resting or elevated SSNA conditions. In Protocol 1 (resting SSNA), subjects received a 10 min sham followed by 10 min of LI4 acupuncture during resting conditions. In Protocol 2 (elevated SSNA), subjects performed 10 min of submaximal intermittent handgrip (2:4s work to rest interval at 37+/-3% of maximal voluntary contraction) during sham and LI4 acupuncture conditions. SSNA (peroneal nerve microneurography), heart rate (ECG), and mean arterial blood pressure (finger photoplethysmography) were measured and normalized to baseline. SSNA, heart rate, and mean arterial blood pressure were not significantly altered during resting conditions (Protocol 1). During handgrip (Protocol 2), SSNA significantly increased with the sham treatment (+15.3+/-8.8, +11.1+/-5.9, and +24.3+/-13.0% at minutes 1, 5, and 10, respectively), while LI4 acupuncture attenuated this increase (-1.6+/-7.6, 0.0+/-4.3, and +2.2+/-11.2% at minutes 1, 5, and 10, respectively). Heart rate and mean arterial blood pressure increased during handgrip (Protocol 2), but no differences were observed between sham and LI4 acupuncture treatments. These results suggest that acupuncture does not affect resting SSNA in healthy subjects, however if SSNA is acutely elevated above baseline levels, acupuncture has the capacity to attenuate the increased SSNA. PMID- 21620781 TI - Assessing multidrug resistance protein 1-mediated function in cancer cell multidrug resistance by scanning electrochemical microscopy and flow cytometry. AB - Cancer cell multidrug resistance is a molecular signature that highly influences the outcome of chemotherapy treatment and for which there is currently no robust method to monitor in vitro its activity. Herein, we demonstrate that ferrocenemethanol (FcCH(2)OH) and its oxidized form ([FcCH(2)OH](+)) affect the redox state of cancer cells. Specifically, the interaction of FcCH(2)OH with the glutathione couple (GSH/GSSG) is shown in human adenocarcinoma cervical cancer cells HeLa and a multidrug resistant variant overexpressing the multidrug resistant associated protein 1 (MRP1) using bioanalytical techniques, such as flow cytometry and fluorescence microscopy. It is further demonstrated that the differential response to FcCH(2)OH in multidrug-resistant cells is in part due to MRP1's unspecific efflux. Scanning electrochemical microscopy confirmed the interaction between FcCH(2)OH and the cells, and the differential response was observed to depend on MRP1 expression. This newly established relation between FcCH(2)OH/[FcCH(2)OH](+), GSH/GSSG and multidrug resistance in human cancer cells enables than the acquisition of scanning electrochemical microscopy images. PMID- 21620782 TI - A randomised controlled trial of breathing modes for adaptive aerosol delivery in children with cystic fibrosis. AB - BACKGROUND: Aerosol delivery is a cornerstone of CF airways disease management. New nebulisers have reduced treatment times by utilising mesh technology for aerosol production. We have evaluated a further modification (target inhalation mode (TIM)) that may reduce treatment delivery times further. METHODS: Following a baseline period on tidal breathing mode (TBM), children with CF on long-term aerosol therapy were randomly allocated to either TIM, which optimises patient inhalations through a direct feedback mechanism, or to continue TBM. The primary outcome was nebuliser treatment times with secondary outcomes being adherence and patient preference. RESULTS: The ten children allocated TIM reduced their mean (SD) treatment times from 6.9(2.9) to 3.7(2.3) minutes (p<0.001). In contrast, treatment times were unchanged in the ten children allocated TBM. Mean adherence was maintained in the TIM group but declined in patients allocated TBM by >5%. All children preferred TIM to TBM. CONCLUSION: TIM reduces nebuliser treatment times and may positively impact on adherence, although longer duration studies are required to examine this. (ISRCTN65617839). PMID- 21620783 TI - Identification of hyperoside metabolites in rat using ultra performance liquid chromatography/quadrupole-time-of-flight mass spectrometry. AB - In this paper, ultra performance liquid chromatography (UPLC)/quadrupole-time-of flight mass spectrometry (QTOF) with automated data analysis software (MetabolynxTM) were applied for fast analysis of hyperoside metabolites in rat after intravenous administration. MS(E) was used for simultaneous acquisition of precursor ion information and fragment ion data at high and low collision energy in one analytical run, which facilitated the fast structural characterization of 12 metabolites in rat plasma, urine and bile. The results indicated that methylation, sulfation and glucuronidation were the major metabolic pathways of hyperoside in vivo, and among them, 3'-O-methyl-hyperoside was confirmed by matching its fragmentation patterns with standard compound. The present study provided important information about the metabolism of hyperoside which will be helpful for fully understanding the mechanism of this compound's action. Furthermore, this work demonstrated the potential of the UPLC/QTOFMS approach using Metabolynx for fast and automated identification of metabolites of natural product. PMID- 21620784 TI - Bilateral pneumothorax has no specific impact upon endoscopic rectal perforation management. PMID- 21620785 TI - Mycobacterium avium subsp paratuberculosis--incidences in milk and milk products, their isolation, enumeration, characterization, and role in human health. AB - Mycobacterium avium subsp paratuberculosis (MAP), excreted in the feces and milk, is reported to be not easily inactivated by pasteurization and thermal treatments as other bacteria infecting humans and animals do. The D values of all MAP strains tested were considerably higher than those published for other pathogens. Culturing techniques for this organism are labor intensive. Although an increasing amount of scientific evidence suggests that this organism can be responsible for at least some cases of Crohn's disease (CD), there is controversy about MAP being a cause of CD in humans. In general, although some studies have described an association between the presence of MAP and CD, the role of Mycobacterium species and MAP in the etiology of this human disease remains unestablished. Although published reports indicate that it may not be completely inactivated by pasteurization of milk, the effectiveness of increasing the time or temperature in the pasteurization process has not been established and hence any potential benefit to human health cannot be determined. This article summarizes the incidences of MAP in milk and milk products with respect to human health and brief discussion of various serological as well as molecular techniques used for their isolation, enumeration, and characterization. PMID- 21620786 TI - Metabolic disorders of fetal life: glycogenoses and mitochondrial defects of the mitochondrial respiratory chain. AB - Two major groups of inborn errors of energy metabolism are reviewed -glycogenoses and defects of the mitochondrial respiratory chain - to see how often these disorders present in fetal life or neonatally. After some general considerations on energy metabolism in the pre- and postnatal development of the human infant, different glycogen storage diseases and mitochondrial encephalomyopathies are surveyed. General conclusions are that: (i) disorders of glycogen metabolism are more likely to cause 'fetal disease' than defects of the respiratory chain; (ii) mitochondrial encephalomyopathies, especially those due to defects of the nuclear genome, are frequent causes of neonatal or infantile diseases, typically Leigh syndrome, but usually do not cause fetal distress; (iii) notable exceptions include mutations in the complex III assembly gene BCS1L resulting in the GRACILE syndrome (growth retardation, aminoaciduria, cholestasis, iron overload, lactic acidosis, and early death), and defects of mitochondrial protein synthesis, which are the 'new frontier' in mitochondrial translational research. PMID- 21620787 TI - Infantile mitochondrial encephalopathy. AB - Individually rare, when taken as a whole, genetic inborn errors of metabolism (IEM) account for a significant proportion of early onset encephalopathy. Prompt diagnosis is crucial to assess appropriate investigation and can sometimes warrant successful therapy. Recent improvements in technology and expansion of knowledge on the biochemical and molecular basis of these disorders allow astute child neurologists and paediatricians to improve the early diagnosis of these genetically determined defects. However, because of rarity and heterogeneity of these disorders, IEM encephalopathies are still a formidable challenge for most physicians. The most frequent cause of childhood IEM encephalopathy is mitochondrial disease, whose biochemical 'signature' is faulty energy supply due to defects of the last component of the oxidative pathways residing within mitochondria, i.e. the mitochondrial respiratory chain. PMID- 21620788 TI - Mastoid osteoma: report of two cases. AB - INTRODUCTION: Mastoid osteoma is a rare benign tumour. In this article, the authors report two new cases of mastoid osteoma and discuss the modalities of diagnosis and management. CASE REPORTS: Both patients presented with a retroauricular mass that had been slowly increasing in size over several years. The patients consulted for the cosmetic deformity induced by the lesion or moderate tenderness. The diagnosis was based on the clinical presentation and non contrast CT. The osteoma was surgically resected in one patient. DISCUSSION/CONCLUSION: Mastoid osteoma is a rare, slowly growing, and usually asymptomatic benign tumour. Diagnosis is based on clinical findings and CT. Surgery is indicated for symptomatic or cosmetically unacceptable osteomas. PMID- 21620789 TI - Reprogramming of mouse and human cells to pluripotency using mature microRNAs. AB - Induced pluripotent stem cells (iPSCs) can be generated from differentiated human and mouse somatic cells using transcription factors such as Oct4, Sox2, Klf4, and c-Myc. It is possible to augment the reprogramming process with chemical compounds, but issues related to low reprogramming efficiencies and, with a number of protocols, residual vector sequences, remain to be resolved. We show here that it is possible to reprogram mouse and human cells to pluripotency by direct transfection of mature double-stranded microRNAs (miRNAs). Our approaches use a combination of mir-200c plus mir-302 s and mir-369 s family miRNAs. Because this reprogramming method does not require vector-based gene transfer, it holds significant potential for biomedical research and regenerative medicine. PMID- 21620790 TI - TLRs innate immunereceptors and Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) CIDR1alpha-driven human polyclonal B-cell activation. AB - Chronic malaria severely affects the immune system and causes polyclonal B-cell activation, as evidenced by the presence of hypergammaglobulinemia, elevated levels of autoantibodies, loss of B-cell memory and the frequent occurrence of Burkitt's lymphomas (BL) in children living in malaria endemic areas. Previous studies have shown that the cysteine-rich interdomain region 1alpha (CIDR1alpha) of the Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) of the FCR3S1.2 strain, subsequently named CIDR1alpha, interacts with B cells partially through the binding to the B-cell receptor (BCR). This interaction leads to an activated phenotype, increased survival, and a low degree of proliferation. CIDR1alpha preferentially activates the memory B-cell compartment, therefore PfEMP1 is considered to act as a polyclonal B-cell activator and its role in memory maintenance has been suggested. In this report, we extend the analysis of the PfEMP1-CIDR1alpha B-cell interaction and demonstrate that PfEMP1-CIDR1alpha increases the expression of TLR7 and TLR10 mRNA transcripts and sensitizes B cells to TLR9 signalling via the MyD88 adaptor molecule. Furthermore, despite its ability to bind to surface Igs, PfEMP1-CIDR1alpha-induced B-cell activation does not seem to proceed through the BCR, since it does not induce Lyn and/or phospho tyrosine mediated signalling pathways. Rather PfEMP1-CIDR1alpha induces the phosphorylation of downstream kinases, such as ERK1/2, p38 and IKBalpha, in human B cells. These findings indicate that PfEMP1-CIDR1alpha induces a persistent activation of B cells, which in turn can contribute to the exhaustion and impairment of B-cell functions during chronic malaria infection. PMID- 21620791 TI - Immuno-rolling circle amplification using a multibinding fusion protein. AB - Ultrasensitive detection of specific, low level proteins in body fluids is particularly challenging. Owing to the extreme sensitivity of the polymerase chain reaction step, the requirements for immuno-rolling circle amplification (immuno-RCA) are much more stringent than for conventional ELISA. Here, we report the development of a rolling circle amplification procedure using multibinding fusion protein to enhance signals of immuno-RCA to detect a cancer biomarker, alpha-fetoprotein (AFP). We successfully avoid the covalent linkage between antibody and DNA or antibody and biotin/streptavidin by introducing a new genetically engineered fusion protein which contains the C2 domain of protein G and biotin acceptor peptide (BAP) which is intended to maintain the biological activity of the antibody. The purified fusion protein retained its binding affinity with IgG and streptavidin after efficient expression in Escherichia coli. Immuno-RCA in combination with BAP-C2 specifically and sensitively detected AFP in a microplate format. Therefore, the sensitivity and convenient nature of this method should contribute to effective signal enhancement in immunoassays for cancer biomarker detection. PMID- 21620792 TI - Periodic acid-Schiff's reagent assay for carbohydrates in a microtiter plate format. AB - Microtiter plate colorimetric assays are widely used for analysis of carbohydrates and glycoconjugates. However, mucins are often not easily detected, as they have low neutral sugar content. We have adapted and optimised the periodic acid-Schiff's reagent (PAS) staining for microtiter plate assay by examining five factors: concentration and volume of periodic acid, oxidation time, volume of Schiff's reagent, and color development time. This assay requires just 25 MUl of sample, utilises standardised Schiff's reagent, and has decreased assay time (140 min to completion). Seventeen monosaccharides (acidic, neutral, basic, phosphorylated, and deoxy) and four disaccharides were assessed. PAS positive carbohydrates (amino, N-acetylamino, deoxy, and certain neutral monosaccharides, and sialic acids) responded linearly within a 10-100 nmol range approximately, which varied for each carbohydrate. The assay response for fetuin and porcine gastric mucin (PGM) was linear up to 150 MUg (highest concentration tested), with no response from nonglycosylated protein. A lower response for asialofetuin was observed, but desialylated PGM preparations were similar or higher in response than their sialylated counterparts. The simplicity and low sample consumption of this method make it an excellent choice for screening or quantitation of chromatographic fractions containing carbohydrates and glycoconjugates, especially in the case of mucins. PMID- 21620793 TI - Development of a novel assay for human tyrosyl DNA phosphodiesterase 2. AB - Tyrosyl DNA phosphodiesterase 2 (TDP2), a newly discovered enzyme that cleaves 5' phosphotyrosyl bonds, is a potential target for chemotherapy. TDP2 possesses both 3'- and 5'-tyrosyl-DNA phosphodiesterase activity, which is generally measured in a gel-based assay using 3'- and 5'-phosphotyrosyl linkage at the 3' and 5' ends of an oligonucleotide. To understand the enzymatic mechanism of this novel enzyme, the gel-based assay is useful, but this technique is cumbersome for TDP2 inhibitor screening. For this reason, we have designed a novel assay using p nitrophenyl-thymidine-5'-phosphate (T5PNP) as a substrate. This assay can be used in continuous colorimetric assays in a 96-well format. We compared the salt and pH effect on product formation with the colorimetric and gel-based assays and showed that they behave similarly. Steady-state kinetic studies showed that the 5' activity of TDP2 is 1000-fold more efficient than T5PNP. Tyrosyl DNA phosphodiesterase 1 (TDP1) and human AP-endonuclease 1 (APE1) could not hydrolyze T5PNP. Sodium orthovanadate, a known inhibitor of TDP2, inhibits product formation from T5PNP by TDP2 (IC(50)=40 mM). Our results suggest that this novel assay system with this new TDP2 substrate can be used for inhibitor screening in a high-throughput manner. PMID- 21620794 TI - Carotenoids inhibit proliferation and regulate expression of peroxisome proliferators-activated receptor gamma (PPARgamma) in K562 cancer cells. AB - As one of the main micronutrients in vegetables and fruit carotenoids are almost daily intaken in significant quantity. Although the pharmacological roles of carotenoids in the prevention and reduction of cancer incidence have received more and more attention, the exact molecular mechanisms underlying anticancer effects of carotenoids remain unclear yet. Activated peroxisome proliferator activated receptor gamma (PPARgamma) plays an inhibitory role in cancer cell proliferation and growth. Involvement of PPARgamma in the growth inhibition of leukemia K562 cells by carotenoids was investigated in the present study. The results demonstrated that beta-carotene, astaxanthin, capsanthin, and bixin inhibited the proliferation and decreased the viability of leukemia K562 cells in dose- and time-dependent manners, induced cell apoptosis, and interfered with cell cycle progression. Pretreatment with GW9662, a potent antagonist of PPARgamma, partly attenuated the inhibition of K562 cell proliferation by the four carotenoids at 8MUM. These carotenoids up-regulated the expression of PPARgamma and p21 and down-regulated the expression of cyclin D1 in a dose dependent manner. In addition, beta-carotene, astaxanthin, capsanthin and bixin also up-regulated the expression of Nrf2, an important transcription factor in Keap1-Nrf2/EpRE/ARE signaling pathway. It appears to us that PPARgamma signaling pathways and Keap1-Nrf2/EpRE/ARE signaling pathway were involved in the inhibition of K562 cell proliferation by carotenoids and the up-regulation of PPARgamma expression at least partly contributed to the antiproliferative effects of beta-carotene, astaxanthin, capsanthin, and bixin on K562 cells. PMID- 21620795 TI - The D-channel of cytochrome oxidase: an alternative view. AB - The D-pathway in A-type cytochrome c oxidases conducts protons from a conserved aspartate on the negatively charged N-side of the membrane to a conserved glutamic acid at about the middle of the membrane dielectric. Extensive work in the past has indicated that all four protons pumped across the membrane on reduction of O(2) to water are transferred via the D-pathway, and that it is also responsible for transfer of two out of the four "chemical protons" from the N side to the binuclear oxygen reduction site to form product water. The function of the D-pathway has been discussed in terms of an apparent pK(a) of the glutamic acid. After reacting fully reduced enzyme with O(2), the rate of formation of the F state of the binuclear heme-copper active site was found to be independent of pH up to pH~9, but to drop off at higher pH with an apparent pK(a) of 9.4, which was attributed to the glutamic acid. Here, we present an alternative view, according to which the pH-dependence is controlled by proton transfer into the aspartate residue at the N-side orifice of the D-pathway. We summarise experimental evidence that favours a proton pump mechanism in which the proton to be pumped is transferred from the glutamic acid to a proton-loading site prior to proton transfer for completion of oxygen reduction chemistry. The mechanism is discussed by which the proton-pumping activity is decoupled from electron transfer by structural alterations of the D-pathway. This article is part of a Special Issue entitled: Allosteric cooperativity in respiratory proteins. PMID- 21620796 TI - Reprint of: physiology of PSI cyclic electron transport in higher plants. AB - Having long been debated, it is only in the last few years that a concensus has emerged that the cyclic flow of electrons around Photosystem I plays an important and general role in the photosynthesis of higher plants. Two major pathways of cyclic flow have been identified, involving either a complex termed NDH or mediated via a pathway involving a protein PGR5 and two functions have been described-to generate ATP and to provide a pH gradient inducing non-photochemical quenching. The best evidence for the occurrence of the two pathways comes from measurements under stress conditions-high light, drought and extreme temperatures. In this review, the possible relative functions and importance of the two pathways is discussed as well as evidence as to how the flow through these pathways is regulated. Our growing knowledge of the proteins involved in cyclic electron flow will, in the future, enable us to understand better the occurrence and diversity of cyclic electron transport pathways. This article is part of a Special Issue entitled: Regulation of Electron Transport in Chloroplasts. PMID- 21620797 TI - Modulation of N-type Ca2+ currents by moxonidine via imidazoline I1 receptor activation in rat superior cervical ganglion neurons. AB - Moxonidine, an imidazoline deriviatives, suppress the vasopressor sympathetic outflow to produce hypotension. This effect has been known to be mediated in part by suppressing sympathetic outflow via acting imidazoline I(1) receptors (IR(1)) at postganglionic sympathetic neurons. But, the cellular mechanism of IR(1) induced inhibition of noradrenaline (NA) release is still unknown. We therefore, investigated the effect of IR(1) activation on voltage-dependent Ca(2+) channels which is known to play an pivotal role in regulating NA in rat superior cervical ganglion (SCG) neurons, using the conventional whole-cell patch-clamp method. In the presence of rauwolscine (3 MUMU), which blocks alpha(2)-adrenoceptor (R(alpha2)), moxonidine inhibited voltage-dependent Ca(2+) current (I(Ca)) by about 30%. This moxonidine-induced inhibition was almost completely prevented by efaroxan (10 MUMU) which blocks IR(1) as well as R(alpha2). In addition, omega conotoxin (CgTx) GVIA (1 MUMU) occluded moxonidine-induced inhibition of I(Ca), but, moxonidine-induced I(Ca) inhibition was not affected by pertussis toxin (PTX) nor shows any characteristics of voltage-dependent inhibition. These data suggest that moxonidine inhibit voltage-dependent N-type Ca(2+) current (I(Ca-N)) via activating IR(1). Finally, moxonidine significantly decreased the frequency of AP firing in a partially reversible manner. This inhibition of AP firing was almost completely occluded in the presence of omega-CgTx. Taken together, our results suggest that activation of IR(1) in SCG neurons reduced I(Ca-N) in a PTX and voltage-insensitive pathway, and this inhibition attenuated repetitive AP firing in SCG neurons. PMID- 21620798 TI - The microcephaly gene aspm is involved in brain development in zebrafish. AB - MCPH is a neurodevelopmental disorder characterized by a global reduction in cerebral cortical volume. Homozygous mutation of the MCPH5 gene, also known as ASPM, is the most common cause of the MCPH phenotype. To elucidate the roles of ASPM during embryonic development, the zebrafish aspm was identified, which is specifically expressed in proliferating cells in the CNS. Morpholino-mediated knock-down of aspm resulted in a significant reduction in head size. Furthermore, aspm-deficient embryos exhibited a mitotic arrest during early development. These findings suggest that the reduction in brain size in MCPH might be caused by lack of aspm function in the mitotic cell cycle and demonstrate that the zebrafish can provide a model system for congenital diseases of the human nervous system. PMID- 21620799 TI - Nuclear translocation of the cytoskeleton-associated protein, smALP, upon induction of skeletal muscle differentiation. AB - The skALP isoform has been shown to play a critical role in actin organization and anchorage within the Z-discs of skeletal muscles, but no data is available on the function of the smALP isoform in skeletal muscle cells. Here, we show that upon induction of differentiation a nuclear translocation of smALP from the cytoplasm to the nucleus of C2C12 myoblasts, concomitant to an up-regulation of the protein expression, occurs in parallel with the nuclear accumulation of myogenin. Moreover, we demonstrate that the LIM domain of smALP is essential for the nuclear translocation of the protein. PMID- 21620800 TI - CPEB1 modulates lipopolysaccharide-mediated iNOS induction in rat primary astrocytes. AB - Upon CNS damage, astrocytes undergo a series of biological changes including increased proliferation, production of inflammatory mediators and morphological changes, in a response collectively called reactive gliosis. This process is an essential part of the brains response to injury, yet much is unknown about the molecular mechanism(s) that induce these changes. In this study, we investigated the role of cytoplasmic polyadenylation element binding protein 1 (CPEB1) in the regulation of inflammatory responses in a model of reactive gliosis, lipopolysaccharide-stimulated astrocytes. CPEB1 is an mRNA-binding protein recently shown to be expressed in astrocytes that may play a role in astrocytes migration. After LPS stimulation, the expression and phosphorylation of CPEB1 was increased in rat primary astrocytes in a JNK-dependent process. siRNA-induced knockdown of CPEB1 expression inhibited the LPS-induced up-regulation of iNOS as well as NO and ROS production, a hallmark of immunological activation of astrocytes. The results from the study suggest that CPEB1 is actively involved in the regulation of inflammatory responses in astrocytes, which might provide new insights into the regulatory mechanism after brain injury. PMID- 21620801 TI - Plural assay systems derived from different cell lines and hepatitis C virus strains are required for the objective evaluation of anti-hepatitis C virus reagents. AB - Persistent hepatitis C virus (HCV) infection causes chronic liver diseases and is a global health problem. HuH-7 hepatoma-derived cells are widely used as the only cell-based HCV replication system for HCV research, including drug assays. Recently, using different hepatoma Li23-derived cells, we developed an HCV drug assay system (ORL8), in which the genome-length HCV RNA (O strain of genotype 1b) encoding renilla luciferase replicates efficiently. In this study, using the HuH 7-derived OR6 assay system that we developed previously and the ORL8 assay system, we evaluated 26 anti-HCV reagents, which other groups had reported as anti-HCV candidates using HuH-7-derived assay systems other than OR6. The results revealed that more than half of the reagents showed different anti-HCV activities from those in the previous studies, and that anti-HCV activities evaluated by the OR6 and ORL8 assays were also frequently different. In further evaluation using the HuH-7-derived AH1R assay system, which was developed using the AH1 strain of genotype 1b, several reagents showed different anti-HCV activities in comparison with those evaluated by the OR6 and ORL8 assays. These results suggest that the different activities of anti-HCV reagents are caused by the differences in cell lines or HCV strains used for the development of assay systems. Therefore, we conclude that plural HCV assay systems developed using different cell lines or HCV strains are required for the objective evaluation of anti-HCV reagents. PMID- 21620802 TI - Prolyl oligopeptidase participates in cell cycle progression in a human neuroblastoma cell line. AB - Prolyl oligopeptidase (POP) is a post-proline cleaving enzyme, which is widely distributed in various organs, with high levels in the brain. In this study, we investigated the effects of a selective POP inhibitor, 3-({4-[2-(E) styrylphenoxy]butanoyl}-l-4-hydroxyprolyl)-thiazolidine (SUAM-14746), on the growth of NB-1 human neuroblastoma cells. SUAM-14746 treatment for 24-72 h suppresses the growth of NB-1 cells without cell death in a dose-dependent manner (10-60 MUM). Similar suppressive effects were observed with another POP inhibitor benzyloxycarbonyl-thioprolyl-thioprolinal. The SUAM-14746-induced growth inhibition in NB-1 cells was associated with pronounced G(0)/G(1) arrest and reduced levels of phosphorylated retinoblastoma protein (pRb), cyclin E, and cyclin dependent kinase (CDK) 2, and increased levels of the CDK inhibitor p27(kip1) and the tumor suppressor p53. SUAM-14746 also induced transient inhibition of S and G(2)/M phase progression, which was correlated with retardation of the decrease in the levels of cyclins A and B. Moreover, RNAi mediated knockdown of POP also led to inhibition of NB-1 cell growth and the effect was accompanied by G(0)/G(1) arrest. These results indicate that POP is a part of the machinery that controls the cell cycle. PMID- 21620803 TI - Analysis of deoxyribonucleotide pools in human cancer cell lines using a liquid chromatography coupled with tandem mass spectrometry technique. AB - Endogenous ribonucleotides and deoxyribonucleotides play a critical role in cell function, and determination of their levels is of fundamental importance in understanding key cellular processes involved in energy metabolism and molecular and biochemical signaling pathways. In this study, we determined the respective ribonucleotide and deoxyribonucleotide pool sizes in different human cell lines using a simple sample preparation method and LC/MS/MS. This assay was used to determine alterations in deoxyribonucleotide pools in human pancreatic PANC1 cells in response to hypoxia and to treatment with either hydroxyurea or aphidicolin. The levels of all deoxyribonucleotide metabolites decreased with hypoxia treatment, except for dUMP, which increased by two-fold. This LC/MS/MS assay is simple, fast, and sensitive, and it represents a significant advance over previously published methodologies. PMID- 21620804 TI - The four cysteine residues in the second extracellular loop of the human adenosine A2B receptor: role in ligand binding and receptor function. AB - The adenosine A(2B) receptor is of considerable interest as a new drug target for the treatment of asthma, inflammatory diseases, pain, and cancer. In the present study we investigated the role of the cysteine residues in the extracellular loop 2 (ECL2) of the receptor, which is particularly cysteine-rich, by a combination of mutagenesis, molecular modeling, chemical and pharmacological experiments. Pretreatment of CHO cells recombinantly expressing the human A(2B) receptor with dithiothreitol led to a 74-fold increase in the EC(50) value of the agonist NECA in cyclic AMP accumulation. In the C78(3.25)S and the C171(45.50)S mutant high affinity binding of the A(2B) antagonist radioligand [(3)H]PSB-603 was abolished and agonists were virtually inactive in cAMP assays. This indicates that the C3.25-C45.50 disulfide bond, which is highly conserved in GPCRs, is also important for binding and function of A(2B) receptors. In contrast, the C166(45.45)S and the C167(45.46)S mutant as well as the C166(45.45)S-C167(45.46)S double mutant behaved like the wild-type receptor, while in the C154(45.33)S mutant significant, although more subtle effects on cAMP accumulation were observed - decrease (BAY60-6583) or increase (NECA) - depending on the structure of the investigated agonist. In contrast to the X-ray structure of the closely related A(2A) receptor, which showed four disulfide bonds, the present data indicate that in the A(2B) receptor only the C3.25-C45.50 disulfide bond is essential for ligand binding and receptor activation. Thus, the cysteine residues in the ECL2 of the A(2B) receptor not involved in stabilization of the receptor structure may have other functions. PMID- 21620806 TI - Potentiation of analgesic efficacy but not side effects: co-administration of an alpha4beta2 neuronal nicotinic acetylcholine receptor agonist and its positive allosteric modulator in experimental models of pain in rats. AB - Positive modulation of the neuronal nicotinic acetylcholine receptor (nAChR) alpha4beta2 subtype by selective positive allosteric modulator NS-9283 has shown to potentiate the nAChR agonist ABT-594-induced anti-allodynic activity in preclinical neuropathic pain. To determine whether this benefit can be extended beyond neuropathic pain, the present study examined the analgesic activity and adverse effect profile of co-administered NS-9283 and ABT-594 in a variety of preclinical models in rats. The effect of the combined therapy on drug-induced brain activities was also determined using pharmacological magnetic resonance imaging. In carrageenan-induced thermal hyperalgesia, co-administration of NS 9283 (3.5 MUmol/kg, i.p.) induced a 6-fold leftward shift of the dose-response of ABT-594 (ED(50)=26 vs. 160 nmol/kg, i.p.). In the paw skin incision model of post operative pain, co-administration of NS-9283 similarly induced a 6-fold leftward shift of ABT-594 (ED(50)=26 vs. 153 nmol/kg). In monoiodo-acetate induced knee joint pain, co-administration of NS-9283 enhanced the potency of ABT-594 by 5 fold (ED(50)=1.0 vs. 4.6 nmol/kg). In pharmacological MRI, co-administration of NS-9283 was shown to lead to a leftward shift of ABT-594 dose-response for cortical activation. ABT-594 induced CNS-related adverse effects were not exacerbated in presence of an efficacious dose of NS-9283 (3.5 MUmol/kg). Acute challenge of NS-9283 produced no cross sensitization in nicotine-conditioned animals. These results demonstrate that selective positive allosteric modulation at the alpha4beta2 nAChR potentiates nAChR agonist-induced analgesic activity across neuropathic and nociceptive preclinical pain models without potentiating ABT-594-mediated adverse effects, suggesting that selective positive modulation of alpha4beta2 nAChR by PAM may represent a novel analgesic approach. PMID- 21620805 TI - Structure-activity relationships for the interactions of 2'- and 3'-(O)-(N methyl)anthraniloyl-substituted purine and pyrimidine nucleotides with mammalian adenylyl cyclases. AB - Membranous adenylyl cyclases (ACs) play a key role in signal transduction and are promising drug targets. In previous studies we showed that 2',3'-(O)-(N methylanthraniloyl) (MANT)-substituted nucleotides are potent AC inhibitors. The aim of this study was to provide systematic structure-activity relationships for 21 (M)ANT-substituted nucleotides at the purified catalytic AC subunit heterodimer VC1:IIC2, the VC1:VC1 homodimer and recombinant ACs 1, 2 and 5. (M)ANT-nucleotides inhibited fully activated VC1:IIC2 in the order of affinity for bases hypoxanthine>uracil>cytosine>adenine~guanine?xanthine. Omission of a hydroxyl group at the 2' or 3'-position reduced inhibitor potency as did introduction of a gamma-thiophosphate group or omission of the gamma-phosphate group. Substitution of the MANT-group by an ANT-group had little effect on affinity. Although all nucleotides bound to VC1:IIC2 similarly according to the tripartite pharmacophore model with a site for the base, the ribose, and the phosphate chain, nucleotides exhibited subtle differences in their binding modes as revealed by fluorescence spectroscopy and molecular modelling. MANT nucleotides also differentially interacted with the VC1:VC1 homodimer as assessed by fluorescence spectroscopy and modelling. Similar structure-activity relationships as for VC1:IIC2 were obtained for recombinant ACs 1, 2 and 5, with AC2 being the least sensitive AC isoform in terms of inhibition. Overall, ACs possess a broad base-specificity with no preference for the "cognate" base adenine as verified by enzyme inhibition, fluorescence spectroscopy and molecular modelling. These properties of ACs are indicative for ligand-specific conformational landscapes that extend to the VC1:VC1 homodimer and should facilitate development of non-nucleotide inhibitors. PMID- 21620807 TI - The role of the glutathione system in seizures induced by diphenyl diselenide in rat pups. AB - The present study investigated the role of the glutathione system in seizures induced by diphenyl diselenide (PhSe)(2) (50 mg/kg) in rat pups (post natal day, 12-14). Reduced glutathione (GSH) (300 nmol/site; i.c.v.), administered 20 min before (PhSe)(2), abolished the appearance of seizures, protected against the inhibition of catalase and delta-aminolevulinic dehydratase (delta-ALA-D) activities and increased glutathione peroxidase (GPx) activity induced by (PhSe)(2). Administration of l-buthionine sulfoximine (BSO, a GSH-depleting compound) (3.2 MUmol/site; i.c.v.) 24h before (PhSe)(2) increased the percentage (42-100%) of rat pups which had seizure episodes, reduced the onset for the first convulsive episode. In addition, BSO increased thiobarbituric acid reactive species (TBARS) levels and decreased GSH content, catalase, delta-ALA-D and Na(+), K(+)-ATPase activities. Treatment with sub effective doses of GSH (10 nmol/site) and d-2-amino-7-phosphonoheptanoic acid (AP-7, an antagonist of the glutamate site at the NMDA receptor; 5mg/kg, i.p.) abolished the appearance of seizures induced by (PhSe)(2) in rat pups. Sub effective doses of GSH and kynurenic acid (an antagonist of strychnine-insensitive glycine site at the NMDA receptor; 40 mg/kg, i.p.) were also able in abolishing the appearance of seizures induced by (PhSe)(2). In conclusion, administration of GSH protected against seizure episodes induced by (PhSe)(2) in rat pups by reducing oxidative stress and, at least in part, by acting as an antagonist of glutamate and glycine modulatory sites in the NMDA receptor. PMID- 21620808 TI - Validation of an isotope dilution gas chromatography-mass spectrometry method for analysis of 7-oxygenated campesterol and sitosterol in human serum. AB - High dose daily intake of plant sterols decreases the uptake of cholesterol in the intestine by competitive mechanisms and thus leads to reduced serum levels of total and LDL-cholesterol. By this, the commercialization of plant sterol enriched 'functional food' products is rapidly increasing. Subjects using these kinds of diet present a duplication of their serum plant sterol levels after long term intake. In analogy to cholesterol, plant sterols such as campesterol and sitosterol can be oxidized to oxyphytosterols and these may counteract the primary anti-atherosclerotic action of cholesterol lowering. In order to investigate the whole spectrum of the consequences following high plant sterol intake a highly sensitive and specific isotope dilution gas chromatography-mass spectrometry method for the analysis of 7-oxygenated campesterol/sitosterol in trace amounts in human serum is presented in this paper. The validation was based on limits for detection and quantification, recovery, precision and minimization of autoxidation during work-up. Our results show an overall coefficient of variation <=10% for the precision. The lowest limits for detection and quantification for 7alpha-hydroxy-campesterol were 7 pg/mL and 23 pg/mL, respectively. Data for overall sum recovery ranged from 92% to 115%. We practically used this method for analysis of oxyphytosterols simultaneously with plant sterol concentrations in serum from healthy volunteers. Sixteen subjects were treated with plant sterol enriched margarine (3 g/day) for 28 days. The results showed a significant increase of the oxyphytosterol 7beta-hydroxy sitosterol from 1.19+/-0.54 (before intake) to 2.24+/-1.24 ng/mL (mean+/-SD; +86.7%; P=0.007) after intake of the margarine. There was a highly significant correlation between the serum levels of campesterol and the sum of 7-oxygenated campesterol (R(2)=0.915; P<0.001) and sitosterol and the sum of 7-oxygenated sitosterol (R(2)=0.915; P<0.001). We can conclude from this study that the analytic method is well suited for detection of OPS, even at trace amounts. PMID- 21620809 TI - The clinical significance of a urinary B-type natriuretic peptide assay for the diagnosis of heart failure. AB - BACKGROUND: The purpose of this study was to investigate the applicability of urinary B-type natriuretic peptide (BNP) levels to the diagnosis and prognosis of heart failure (HF) by comparing urinary and plasma BNP levels. METHODS: Urinary and plasma BNP levels of 160 patients with HF classifiable into NYHA stages I, II, III, or IV, and 30 healthy control subjects were measured using the microparticle enzyme immunoassay (MEIA). The heart function of each patient was examined by ultrasonic cardiogram and classified according to the NYHA standard. RESULTS: The levels of urinary and plasma BNP among patients in the HF group were significantly higher than those in the control group. Urinary BNP levels were correlated with plasma BNP levels and NYHA grades of HF and were negatively correlated with LVEF. According to the Cox model multivariate regression analyses on age, gender, heart function class, LVEF, and urinary BNP among patients with post-treatment cardiac events, age and the urinary BNP level were predictors for the independent post-treatment cardiac events. CONCLUSIONS: Measurement of urinary BNP is applicable in clinical diagnosis and prognosis of HF; it provides similar accuracy to plasma BNP in the detection of HF, and it has the advantage of being a noninvasive test. PMID- 21620810 TI - Plasma adiponectin as an independent predictor of early death after acute intracerebral hemorrhage. AB - BACKGROUND: Hyperadiponectinemia or hypoadiponectinemia is associated with different diseases. There is a paucity of data on circulating plasma adiponectin concentrations in human intracerebral hemorrhage (ICH). We investigated the plasma adiponectin concentrations in patients with intracerebral hemorrhage, and analyzed the correlation of adiponectin with the severity of brain injury and early mortality after ICH. METHODS: Thirty controls and 86 patients with acute ICH were included. Plasma samples were obtained on admission and at days 1, 2, 3, 5, and 7 after ICH. Their concentrations were measured by enzyme-linked immunosorbent assay. RESULTS: After ICH, plasma adiponectin level of the patients increased immediately within 6h, peaked within 24h, plateaued at day 2, and decreased gradually thereafter. It was substantially higher than that in the controls in a period of 7 days. A multivariate analysis showed plasma adiponectin level was an independent predictor for 1-week mortality (odds ratio, 1.199; 95% CI: 1.035-1.389; P=0.015) and that it was associated with Glasgow coma scale (GCS) score (t=-3.596, P=0.001) and plasma C-reactive protein level (t=4.194, P<0.001). A receiver operating characteristic curve identified that a plasma adiponectin level >16.4 MUg/ml predicted the 1-week mortality of patients with a sensitivity of 65.6% and a specificity of 90.7% (AUC, 0.789; 95% CI: 0.688 0.870). The predictive value of adiponectin concentration was significantly lower than that of GCS score (P=0.007) and hematoma volume (P=0.022). Adiponectin could not improve the predictive values of GCS score (P=0.317) and hematoma volume (P=0.226). CONCLUSIONS: Adiponectin is an independent indicator of early death and may play an anti-inflammatory role after intracerebral hemorrhage. PMID- 21620811 TI - Decreased serum brain-derived neurotrophic factor concentration in young nonobese subjects with low insulin sensitivity. AB - OBJECTIVE: Insulin resistance and type 2 diabetes are associated with an increased risk of neurodegenerative diseases. Decreased brain-derived neurotrophic factor (BDNF) levels might play a role in the pathogenesis of neuropsychiatric disorders. The aim of our study was to estimate serum BDNF concentration in nonobese women divided into subgroups according to their insulin sensitivity. DESIGN AND METHODS: We studied 46 young, healthy, nonobese women. Insulin sensitivity was estimated with the euglycemic-hyperinsulinemic clamp technique. Then, participants were divided into subgroups of high (mean, 12.79+/ 2.01mg/kg fat-free mass/min) and low insulin sensitivity (mean, 7.33+/-1.66mg/kg fat-free mass/min). RESULTS: We observed decreased serum BDNF concentration in women with low insulin sensitivity in comparison to high insulin sensitivity group (3306.11+/-603.10 vs 4141.91+/-755.37pg/mL, p=0.001). Serum BDNF was positively related to insulin sensitivity (r=0.43, p=0.003). This correlation remained significant after adjustment for other estimated parameters. CONCLUSIONS: Serum BDNF is decreased in young nonobese women with low insulin sensitivity. Early detection and prevention of insulin resistance might be useful in the prevention of neurodegenerative disorders. PMID- 21620812 TI - Reference intervals of serum lipid profile in healthy Indian school children and adolescents. AB - OBJECTIVE: To develop age and sex specific reference intervals of serum lipid parameters in healthy Indian children in the age group of 6-17 years. PARTICIPANTS: Study subjects were selected from the schools of Delhi. Children with BMI either in overweight or obese category were excluded to generate reference intervals. The final analysis included 3076 children with BMI in normal range for age. RESULTS: The mean+/-SD, 3rd, 10th, 25th, 50th, 75th, 90th and 97th centile values of lipid parameters are presented. All these parameters were significantly higher in girls than in boys. About 89.2% subjects had total cholesterol in acceptable range while 9.1% had borderline elevated and 1.5% had elevated TC. Optimal serum triglyceride levels were seen in 45.1% subjects while optimal levels of HDL cholesterol were seen in 0.3% subjects only. CONCLUSIONS: Reference intervals of serum lipid parameters for healthy Indian children (6-17 years) are presented. PMID- 21620813 TI - Polymorphisms of pon1 and pon2 genes in hemodialyzed patients. AB - OBJECTIVES: Q192R, L55M and -108C>T polymorphisms of pon1 gene affect PON1 paraoxonase activity while S311C polymorphism of pon2 gene might be associated with coronary heart disease. The aims of this study were to determine the frequencies of Q192R, L55M, -108C>T and S311C polymorphisms in hemodialyzed patients and to examine the relationship between pon1 gene polymorphisms and PON1 paraoxonase activity in those patients. DESIGN AND METHODS: The study included 238 control subjects and 263 hemodialyzed patients. RESULTS: PON1 paraoxonase activity was lower in patients. Genotype frequencies were different between two compared groups only for L55M polymorphism, with control group having higher frequency of MM genotype. Polymorphisms of pon1 gene were associated with significant variation in PON1 paraoxonase activity in both study groups. CONCLUSION: Our results suggest that Q192R, L55M and -108C>T polymorphisms are not by itself the causal factors leading to the lower PON1 paraoxonase activity in hemodialyzed patients. PMID- 21620814 TI - Establishing and evaluating pediatric thyroid reference intervals on the Roche Modular Analytics E 170 using computational statistics and data-mining techniques. AB - OBJECTIVES: This study determined age specific, free thyroxine and thyroid stimulating hormone reference intervals for a pediatric population. METHODS: The data set was partitioned into age groups and the Tukey method was used to identify outliers for removal. The Harris and Boyd algorithm was used to determine if groups could be merged. Reference intervals were calculated using both non-parametric and robust methods. Boot-strapping was used to determine the 90% confidence intervals surrounding the upper and lower limits of each reference interval. These population based reference intervals were then compared to the manufactures suggested reference intervals. RESULTS: This study provides non parametric and robust reference intervals with associated 90% confidence intervals for free thyroxine and thyroid stimulating hormone as measured on the Roche Modular Analytics E 170. CONCLUSIONS: The results emphasize the importance of establishing population-based reference intervals for the clinical laboratory. PMID- 21620815 TI - Robust and efficient genotyping of factor V and prothrombin alleles using amplicon high-resolution melting and synthetic DNA controls: validation of the test with three different commercial chemistries. AB - OBJECTIVES: Developing robust HRM (amplicon High Resolution Melting) analysis valid for different commercial reaction mixes, using synthetic control DNA samples and the RotorGeneQ (Qiagen) instrument. DESIGN AND METHODS: 126 samples were analyzed for the presence of the factor Leiden and the 20210G>A prothrombin alleles. The four alleles were cloned and used to prepare synthetic controls. RESULTS: All mutant alleles present in the sample were successfully detected. Genotyping confidence mean was higher than 95%. CONCLUSIONS: Cost effective HRM genotyping is very reliable using synthetic control DNAs and the RotorGenQ instrument. PMID- 21620816 TI - Correlation of matrix metalloproteinases and their inhibitors with hypoxia and angiogenesis in premenopausal patients with adenocarcinoma of the breast. AB - OBJECTIVES: The present study was designed to correlate the expression of proteins regulating invasion and angiogenesis in patients with adenocarcinoma of the breast. DESIGN AND METHOD: Seventy-five premenopausal breast cancer patients histologically categorized as grades I, II and III were chosen for the study. We analyzed the expression of MMP-2, and -9 and their inhibitors TIMP-2 and RECK together with HIF-1alpha and VEGF in tumor, adjacent tissues and serum samples by immunohistochemical and Western blot analysis. RESULTS: The breast tumors analyzed in the present study were characterized by increased expression of MMP 2, -9, HIF-1alpha and VEGF with differential expression patterns of TIMP-2 and downregulation of RECK. CONCLUSIONS: The simultaneous analysis of the expression of these molecular markers is important to understand the intricate network between key molecules involved in invasion and angiogenesis that eventually determines the clinical course of the disease. PMID- 21620817 TI - Impact of contrast enhanced MRI on lymphocyte DNA damage and serum visfatin level. AB - OBJECTIVES: Nephrogenic systemic fibrosis is a novel clinical entity encountered in subjects undergoing contrast enhanced magnetic resonance imaging (MRI). The aim of the present study is to evaluate the impact of contrast enhanced MRI exposure on lymphocyte DNA damage and serum levels of visfatin. DESIGN AND METHODS: Twenty-eight subjects undergoing contrast enhanced hypophysial MRI with omniscan were included in the study. Blood samples were drawn before MRI, after non-contrast MRI and after contrast enhanced MRI from each subject. Lymphocyte DNA damage was analyzed by the alkaline comet assay, whereas serum visfatin level was assessed with enzyme immuno assay. RESULTS: Both lymphocyte DNA damage and serum visfatin levels were statistically significantly increased in samples withdrawn after contrast enhanced MRI compared to samples withdrawn after non contrast enhanced MRI and baseline samples (ANOVA p<0.001, for both). CONCLUSIONS: Findings of the present study revealed that the contrast enhanced MRI is associated with increased lymphocyte DNA damage and increased serum visfatin level. PMID- 21620818 TI - Systematic parameter optimization of a Me(2)SO- and serum-free cryopreservation protocol for human mesenchymal stem cells. AB - Human mesenchymal stem cells (hMSCs) have great potential for clinical therapy and regenerative medicine. One major challenge concerning their application is the development of an efficient cryopreservation protocol since current methods result in a poor viability and high differentiation rates. A high survival rate of cryopreserved cells requires an optimal cooling rate and the presence of cryoprotective agents (CPA) in sufficient concentrations. The most widely used CPA, dimethylsulfoxide (Me(2)SO), is toxic at high concentrations at temperatures >4 degrees C and has harmful effects on the biological functionality of stem cell as well as on treated patients. Thus, this study investigates different combinations of non-cytotoxic biocompatible substances, such as ectoin and proline, as potential CPAs in a systematic parametric optimization study in comparison to Me(2)SO as control and a commercial freezing medium (Biofreeze(r), Biochrom). Using a freezing medium containing a low proline (1%, w/v) and higher ectoin (10%, w/v) amount revealed promising results although the highest survival rate was achieved with the Biofreeze(r) medium. Cryomicroscopic experiments of hMSCs revealed nucleation temperatures ranging from -16 to -25 degrees C. The CPAs, beside Me(2)SO, did not affect the nucleation temperature. In most cases, cryomicroscopy revealed intracellular ice formation (IIF) during the cryopreservation cycle for all cryoprotocols. The occurence of IIF during thawing increased with the cooling rate. In case of hMSC there was no correlation between the rate of IIF and the post-thaw cell survival. After thawing adipogenic differentiation of the stem cells demonstrated cell functionality. PMID- 21620819 TI - Nitric oxide involvement in consolidation, but not retrieval phase of cognitive performance enhanced by atorvastatin in mice. AB - Atorvastatin, a widely-used medication in treatment of hypercholesterolemia, has shown some benefits in treating cognition impairment in Alzheimer's disease. In this study, effects of atorvastatin on spatial recognition memory and the involvement of nitric oxide (NO) has been determined on consolidation and retrieval of memory in a two-trial recognition Y-maze test. Memory was impaired using scopolamine (1mg/kg, i.p.); atorvastatin (1, 5mg/kg, p.o.) was administered, either in presence or in absence of a non-specific NO synthase inhibitor, L-NAME (3, 10mg/kg, i.p.); a specific inducible NO synthase inhibitor, aminoguanidine (100mg/kg, i.p.); and a NO precursor, L-arginine (750 mg/kg, i.p.). RESULTS: 1) atorvastatin (5mg/kg) significantly improved memory performance in a dose-dependent manner on consolidation and retrieval stage of memory in scopolamine-treated mice; 2) the beneficial effects of atorvastatin on memory consolidation was significantly reversed by L-NAME (10mg/kg) and aminoguanidine; 3) L-arginine slightly potentiated the effects of sub-effective dose of atorvastatin (1mg/kg) on memory consolidation; 4) either L-NAME (up to 10mg/kg), or aminoguanidine did not affect the memory improvement by atorvastatin on retrieval stage; 5) the effects of sub-effective dose of atorvastatin (1mg/kg) on retrieval of memory were not potentiated by L-arginine. The present study demonstrates that atorvastatin improves both consolidation and retrieval phases of memory. This effect is affected by NO synthase inhibitors and NO precursor, L arginine, only in memory consolidation phase, but not in retrieval phase. It is concluded that NO might be involved in consolidation of spatial memory improvement by atorvastatin. PMID- 21620820 TI - Ischemic insult induced apoptotic changes in PC12 cells: protection by trans resveratrol. AB - In this study, we determined the protective potential of trans resveratrol against oxygen-glucose deprivation (OGD) induced reactive oxygen species mediated apoptotic damages in PC12 cells. In vitro model of ischemic cerebral stroke was created by keeping cells in an OGD condition for 6h followed by 24h reoxygenation. Cells received biologically safe doses (5, 10, and 25 MUM) of trans resveratrol in the following schedules for 24h prior to OGD; during 6h of OGD; for 24h post OGD and whole treatment group which starts from 24h before OGD and lasted to 24h post OGD. Anti-ischemic potential of trans resveratrol was assessed by measuring the regulation of lipid peroxidation, reactive oxygen species production, glutathione content, and expression (mRNA and protein) of apoptotic markers such as Bax, Bcl(2) and Caspase-3. Hypoxia inducible factor 1alpha (HIF-1alpha) was also assessed to correlate the changes with ischemic injuries. Significant (P<0.05) restoration in lipid peroxidation, reactive oxygen species, and glutathione content were observed following the treatment of trans resveratrol in cells receiving OGD and re-oxygenation. Changes induced by trans resveratrol could be correlated well with alterations in the expression of Bax, Bcl(2), Caspase-3 and HIF-1alpha. These results indicate that trans resveratrol administration attenuates free radical formation and mitochondria mediated apoptosis perhaps by regulating the expressions of Bax, Bcl(2,) and Caspase-3 in PC12 cells receiving OGD and re-oxygenation insult. PMID- 21620821 TI - Hyperthermia conditioned astrocyte-cultured medium protects neurons from ischemic injury by the up-regulation of HIF-1 alpha and the increased anti-apoptotic ability. AB - It has been demonstrated that conditioned medium from astrocytes challenged by in vitro ischemia (oxygen-glucose deprivation, OGD) improved neuronal survival. In addition, preconditioning stimuli can be cross-tolerant, safeguarding against other types of injury. We therefore hypothesized that hyperthermia-conditioned astrocyte-cultured medium (ACM) might also have protective effect on neurons against ischemic injury. The cultured-media, named 38ACM and 40ACM respectively, were collected after astrocytes had been incubated at 38 degrees C or 40 degrees C for 6h, followed by incubation at 37 degrees C for 24h. It was found that ischemia for 6h induced a significant reduction in the number of neuronal cells and cell-viability, and an increase in lactate dehydrogenase (LDH) release and the percentage of apoptotic nuclei in neurons. Pre-treatment with 38ACM or 40ACM for 24h significantly diminished ischemia injury, enhanced cell viability, reduced LDH release and reversed apoptosis. Western blot analysis showed that treatment with 38ACM or 40ACM for 24h led to a significant increase in hypoxia inducible factor-1 (HIF-1) alpha expression. The EMSA demonstrated that the ACM increased the binding activity of HIF-1 in ischemic neurons. The data implied that hyperthermia-conditioned ACM protects neurons from ischemic injury by up regulating HIF-1 alpha, and the increased binding activity of HIF-1 and anti apoptotic ability. PMID- 21620822 TI - Anti-angiogenic effects of the receptor tyrosine kinase inhibitor, pazopanib, on choroidal neovascularization in rats. AB - Neovascularization in the eye is a major cause of irreversible vision loss. The present study was undertaken to determine mechanisms through which pazopanib, a drug that targets multiple receptor tyrosine kinases such as VEGF receptors, inhibits angiogenesis and experimental choroidal neovascularization (CNV). Pazopanib inhibited VEGF expression by retinal pigment epithelium (RPE) cells and choroidal endothelial cells (CEC), decreased VEGF-induced cellular migration in a dose-dependent manner and suppressed extracellular signal-regulated kinase (ERK) 1/-2 phosphorylation. To assess the impact of pazopanib in vivo, CNV was induced in rats by rupturing the Bruch's membrane by laser coagulation. These experiments demonstrated that twice-daily topical eye drop treatment significantly (P<0.001) decreased leakage from photocoagulated lesions by 89.5%. Furthermore, the thickness of the developed CNV lesions was significantly inhibited by 71.7% (P<0.001) in pazopanib-treated eyes, and immunoreactivity of VEGF was lower than in control eyes. Our data suggest that pazopanib is a promising inhibitor of angiogenesis leading to an effective inhibition of CNV development in vivo. This activity can be largely ascribed to the down-regulation of VEGF release in the retina as well as to impaired VEGF-induced signaling and chemotaxis. Using a convenient topical dosing regimen, pazopanib may prove useful for treating a variety of ocular neovascular diseases such as neovascular age-related macular degeneration. PMID- 21620823 TI - Osthol ameliorates fat milk-induced fatty liver in mice by regulation of hepatic sterol regulatory element-binding protein-1c/2-mediated target gene expression. AB - The objective of this study was to examine the therapeutic effect of osthol, an active constituent of Cnidium monnieri (L.) Cusson (Apiaceae), in hyperlipidemic fatty liver mice and investigate the potential mechanism of the osthol treatment. A mouse model with hyperlipidemic fatty liver was induced by orally feeding the fat milk for 4 weeks. The experimental mice were then treated with osthol 10-40 mg/kg for 6 weeks. After oral administration, the mice in the model and medicine treated groups were continuously given the fat milk for 2 weeks again. Whereafter, the lipid levels in serum and liver, hepatic weight coefficient and histopathological evaluation were measured. The sterol regulatory element-binding protein (SREBP)-1c, SREBP-2, fatty acid synthase (FAS), low density lipoprotein (LDL) receptor and cholesterol 7alpha-hydroxylase (CYP7A) mRNA expressions in liver were examined. The results showed that in the osthol-treated groups, the total cholesterol, triglyceride and free fatty acid levels in serum and liver, and the hepatic weight coefficient were gradually decreased with dose. Importantly, the histopathological evaluation of liver specimens demonstrated that osthol might decrease lipid accumulation. Osthol could increase the mRNA expression of CYP7A and decrease the mRNA expressions of SREBP-1c, SREBP-2, FAS and LDL receptor in liver in fat milk-induced fatty liver mice. These results suggested that osthol might exert the therapeutic effect on fat milk-induced fatty liver in mice, by inhibiting hepatic SREBP-1c/2 mRNA expressions and subsequent modulation of SREBP-1c/2-mediated target genes such as FAS, CYP7A and LDL receptor. PMID- 21620824 TI - Resveratrol prevents bradykinin-induced contraction of rat urinary bladders by decreasing prostaglandin production and calcium influx. AB - Resveratrol, a polyphenol found in grapes and peanuts, exerts beneficial effects on a number of diseases of cardiovascular and central nervous system. However, effects of resveratrol on the urinary system have not been fully investigated. In the present study, we examined effects of resveratrol on bradykinin-induced contraction and release of prostaglandin E2 in isolated rat urinary bladders. The effects of resveratrol on contractions induced by several agonists (prostaglandin E2, prostaglandin F2alpha and carbachol) and high K+ were also examined. We found that resveratrol concentration-dependently reduced the bradykinin-induced contraction in the rat urinary bladder preparations. The higher concentration of resveratrol (100 MUM) abolished the bradykinin-induced prostaglandin E2 release. Similar results were obtained when the cyclooxygenase inhibitor indomethacin (10 MUM) was used instead of resveratrol. Resveratrol also attenuated the prostaglandin E2-, prostaglandin F2alpha-, and to a lesser extent carbachol induced contractions. Contractile responses to bradykinin, prostaglandin E2 and carbachol were largely prevented by blockade of Ca2+ channels with diltiazem. Both resveratrol and diltiazem prevented contractions induced by an addition of Ca2+ (2.5- 10 mM) into Ca2+-free/50 mMK+ solution or by 50 mMK+ solution containing normal Ca2+ (2.5 mM). These results suggest that resveratrol prevents bradykinin-induced contractions by attenuating not only the production of prostaglandins but also actions of them. The effect of resveratrol on contractile actions seems to be in part due to inhibition of Ca2+ influx. Because bradykinin plays an important role in pathological conditions of urinary bladder function, resveratrol may exert beneficial effects on the urinary bladder diseases. PMID- 21620825 TI - BJ-B11, a novel Hsp90 inhibitor, induces apoptosis in human chronic myeloid leukemia K562 cells through the mitochondria-dependent pathway. AB - In the past few years heat shock protein 90 (Hsp90) inhibitors have been reported to possess significant antitumor activity. We investigated, for the first time, the antitumor activity of a novel Hsp90 inhibitor 2-(4-acetyloxycyclohexylamino) 4-(3, 6, 6-trimethyl-4-oxo-4, 5, 6, 7-tetrahydro-1H-indazol-1-yl)-benzamide (BJ B11) and the molecular mechanism underlying the apoptosis it induces in human chronic myeloid leukemia K562 cells. The results revealed that BJ-B11 triggered growth inhibition in K562 cells and other malignant cell lines in vitro with only minor toxicity in a normal human cell line. BJ-B11 inhibited the proliferation of K562 cells in a concentration- and time-dependent manner, with IC(50) values of 1.1 +/- 0.2 MUM and 0.4 +/- 0.1 MUM after 48 and 72 h incubations respectively. This most likely results from cell cycle arrest at the G(0)/G(1) phase and the induction of apoptosis. In addition, BJ-B11 degraded the Hsp90 client proteins Bcr-Abl and Akt, induced activation of caspase-9 and caspase-3, and subsequent cleavage of PARP. The caspase signals may originate from mitochondrial dysfunction, which is supported by the finding of cytochrome c release. In addition, inactivation of the Akt signaling pathway may be involved in the process of BJ-B11-induced apoptosis. Taken together, our data provide a putative molecular mechanism for the anticancer effect of BJ-B11 on K562 cells, and suggest a potential application for BJ-B11 in chronic myeloid leukemia therapy. PMID- 21620826 TI - Protective effects of cortex fraxini coumarines against oxonate-induced hyperuricemia and renal dysfunction in mice. AB - The aim of the present study was to investigate the effects of cortex fraxini coumarines esculetin, esculin, fraxetin and fraxin on renal dysfunction and expression abnormality of renal organic ion transporters in hyperuricemic animals. Mice were orally given 250 mg/kg oxonate for seven consecutive days to induce hyperuricemia and renal dysfunction. After 1h of oxonate induction daily, animals were orally treated with esculetin, esculin, fraxetin and fraxin at 20 and 40 mg/kg, respectively. Esculetin, esculin, fraxetin and fraxin significantly decreased serum urate, creatinine and blood urea nitrogen levels and increased urine urate and creatinine excretion in hyperuricemic mice. Esculetin and esculin up-regulated expressions of renal organic anion transporter 1 (mOAT1), organic cation and carnitine transporters (mOCT1-2 and mOCTN1-2), but failed to affect renal glucose transporter 9 (mGLUT9) and urate transporter 1 (mURAT1) in this model. Fraxetin specifically inhibited renal mURAT1, while fraxin extensively interacted with renal mGLUT9, mURAT1, mOAT1 and mOCT1 in hyperuricemic mice. Furthermore, esculetin, fraxetin and fraxin increased mABCG2 mRNA expression and decreased its protein levels in renal apical membrane in hyperuricemic mice. These results indicate that esculetin and esculin have beneficial effects on hyperuricemia and renal dysfunction, resulting in restoration of mOAT1, mOCT1-2 and mOCTN1-2, and fraxetin and fraxin enhance urate excretion partly by inhibiting mURAT1 or mGLUT9 in kidney of hyperuricemic mice. Regulation of mABCG2 by cortex fraxini coumarines may be partly contributed to their beneficial actions. This study provides an evidence to support clinical therapeutic effects of cortex fraxini coumarines on hyperuricemia with renal dysfunction. PMID- 21620827 TI - The labdane diterpene sclareol (labd-14-ene-8, 13-diol) induces apoptosis in human tumor cell lines and suppression of tumor growth in vivo via a p53 independent mechanism of action. AB - The labdane diterpene sclareol has demonstrated significant cytotoxicity against human tumor cell lines and human colon cancer xenografts. Therefore, there is need to elucidate the mode of action of this compound as very little information is known for the anticancer activity of sclareol and other labdane diterpenes, in general. COMPARE analysis of GI(50) values for a number of human cancer cell lines was initially implicated in an effort to assign a putative mechanism of action to the compound. Sclareol-induced cell cycle arrest and apoptosis were assessed by flow cytometry and Western blot analyses. Finally, the anticancer ability of sclareol in vivo was assessed by using human colon cancer xenograft/mouse models. Sclareol arrested in vitro the growth of p53-deficient (HCT116(p53-/-)) human colon cancer cells and subsequently induced apoptosis by activating both caspases-8 and -9. Intraperitoneal administration of liposome encapsulated sclareol at the maximum tolerated dose induced a marked growth suppression of HCT116(p53-/-) tumors established as xenografts in immunodeficient NOD/SCID mice. In conclusion, we demonstrate herein that sclareol kills human tumor cells by inducing arrest at the G(1)-phase of the cell cycle followed by apoptosis that involves activation of caspases-8, -9 and -3 via a p53-independent mechanism. These findings suggest that liposome-encapsulated sclareol possesses chemotherapeutic potential for the treatment of colorectal and other types of human cancer regardless of the p53-status. PMID- 21620828 TI - Fenofibrate increases neuronal vasoconstrictor response in mesenteric arteries from diabetic rats: role of noradrenaline, neuronal nitric oxide and calcitonin gene-related peptide. AB - We evaluated the possible effects of long-term fenofibrate treatment on adrenergic, nitrergic and CGRP-ergic innervation function in mesenteric arteries from streptotozin-induced diabetic rats. We analysed the vasoconstrictor response to electrical field stimulation (EFS) and the effects of the alpha antagonist phentolamine, the calcitonin gene related peptide (CGRP) receptor antagonist CGRP (8-37) and the nitric oxide synthase (NOS) inhibitor L-NAME in segments from untreated and fenofibrate-treated (100 mg/kg/day) diabetic rats. The vasomotor responses to noradrenaline (NA), CGRP and the NO donor sodium nitroprusside (SNP) were analysed, and NA, CGRP, and NO releases were measured. Neuronal NOS (nNOS), phosphorylated nNOS (P-nNOS), and RAMP1 protein expression were also analysed. Fenofibrate enhanced EFS-induced contractions. Phentolamine reduced EFS-induced contractions more in segments from fenofibrate-treated than in untreated rats. Fenofibrate increased vasoconstrictor response to NA and did not modify NA release. L-NAME increased EFS-induced contractions to a higher extent in segments from fenofibrate-treated than untreated rats. Fenofibrate did not change the vasodilator response to SNP but increased EFS-induced nitric oxide release. CGRP (8-37) increased EFS-induced contractions less in segments from fenofibrate treated rats. Fenofibrate increased the vasodilator response to CGRP and reduced CGRP release. P-nNOS and RAMP1 expression were increased in segments from fenofibrate-treated rats, while nNOS expression remained unmodified. Fenofibrate enhances the vasoconstrictor response to EFS in diabetic rats. This effect is the functional result of the modifications of at least: (i) adrenergic function, enhanced by increased sensitivity to noradrenaline; (ii) nitrergic function, enhanced by increased neuronal NO release; and (iii) CGRP function, decreased by a reduction in CGRP release. PMID- 21620829 TI - Molecular mechanism of vasorelaxant and antiatherogenic effects of the statins in the human saphenous vein graft. AB - In this study we aimed to investigate the vasorelaxant and antiatherogenic effects of the statins (fluvastatin and pravastatin) in the human saphenous vein grafts at the molecular level by using histopathologic, pharmacological and immunochemical techniques. The saphenous vein grafts evaluated histopathologically displayed a loss in their endothelium up to a ratio of 30% and set forth indications of functional deterioration. The pharmacological evaluations proved that the relaxation responses induced by fluvastatin and pravastatin were significantly inhibited by nitric oxide synthase inhibitor, N(G) nitro-l-arginine, and cyclooxygenase inhibitor, indomethacin, while these responses were significantly increased by angiotensin converting enzyme inhibitors, captopril and enalapril, and rho kinase inhibitor, Y27632. The results of immunochemical studies are in accordance with the results of the pharmacological studies that the related statins increased the levels of nitric oxide, phospholipase A(2) and they decreased the levels of angiotensin II and active rho kinase. On the other hand mevalonolactone, a substrate of lipid metabolism, failed to change the effects of fluvastatin and pravastatin in the related tissue. The experimental results indicate that activation of nitric oxide synthase and phospholipase A(2)-cyclooxygenase pathway and inhibition of angiotensin converting enzyme and rho kinase may have a role on the effects of fluvastatin and pravastatin in the human saphenous vein grafts. It seems that the vasorelaxant and antiatherogenic effects of the related statins are independent of their lipid lowering mechanism. PMID- 21620830 TI - TIEG1-null tenocytes display age-dependent differences in their gene expression, adhesion, spreading and proliferation properties. AB - The remodeling of extracellular matrix is a crucial mechanism in tendon development and the proliferation of fibroblasts is a key factor in this process. The purpose of this study was to further elucidate the role of TIEG1 in mediating important tenocyte properties throughout the aging process. Wildtype and TIEG1 knockout tenocytes adhesion, spreading and proliferation were characterized on different substrates (fibronectin, collagen type I, gelatin and laminin) and the expression levels of various genes known to be involved with tendon development were analyzed by RT-PCR. The experiments revealed age-dependent and substrate dependent properties for both wildtype and TIEG1 knockout tenocytes. Taken together, our results indicate an important role for TIEG1 in regulating tenocytes adhesion, spreading, and proliferation throughout the aging process. Understanding the basic mechanisms of TIEG1 in tenocytes may provide valuable information for treating multiple tendon disorders. PMID- 21620831 TI - Dose-response relationship for alpha-tocopherol prevention of ultraviolet radiation induced cataract in rat. AB - The purpose of this study is to establish the dose response relationship for alpha-tocopherol protection of ultraviolet radiation (UVR) induced cataract in the rat. Four groups of 20 six-week-old albino Sprague Dawley rats received 5, 25, 50, and 100 IU/day alpha-tocopherol, whilst another group of 20 rats without any alpha-tocopherol feeding was the control group. After 4 weeks of feeding, each rat was unilaterally exposed to 8 kJ/m(2) UVR-300 nm for 15 min. At 1 week after exposure, the rats were sacrificed and lens light scattering was measured quantitatively. Lens total reduced (GSH) and oxidized (GSSG) glutathione; glutathione reductase (GR) and peroxidase (GPx) were determined spectrophotometrically. The UVR-exposed lenses in the alpha-tocopherol fed groups developed superficial cataract, whereas lenses in the control group developed cortical and equatorial opacities. Light scattering in lenses from the alpha tocopherol-supplemented rats was lower than in lenses from the control group. The difference of light scattering between the exposed and contralateral non-exposed lens decreased with increasing doses of alpha-tocopherol to an asymptote level. UVR-exposure caused a significant depletion of lens GSH in rats without or at low alpha-tocopherol supplementation. The depletion of GSH became less with higher alpha-tocopherol supplementation. There was no detectable difference in lens GSSG, GR or GPx at any level of alpha-tocopherol supplementation. Orally administered alpha-tocopherol dose dependently protects against UVR-induced cataract. The protection is associated with an alpha-tocopherol dose-dependent GSH depletion secondary to UVR exposure. UVR-induced light scattering only occurs if the GSH depletion exceeds a threshold. PMID- 21620832 TI - Hind limb muscle atrophy precedes cerebral neuronal degeneration in G93A-SOD1 mouse model of amyotrophic lateral sclerosis: a longitudinal MRI study. AB - Amyotrophic lateral sclerosis (ALS) is a progressive, fatal, neurodegenerative disorder caused by the degeneration of motor neurons in the CNS, which results in complete paralysis of skeletal muscles. Recent experimental studies have suggested that the disease could initiate in skeletal muscle, rather than in the motor neurons. To establish the timeframe of motor neuron degeneration in relation to muscle atrophy in motor neuron disease, we have used MRI to monitor changes throughout disease in brain and skeletal muscle of G93A-SOD1 mice, a purported model of ALS. Longitudinal MRI examination of the same animals indicated that muscle volume in the G93A-SOD1 mice was significantly reduced from as early as week 8 of life, 4 weeks prior to clinical onset. Progressive muscle atrophy from week 8 onwards was confirmed by histological analysis. In contrast, brain MRI indicated that neurodegeneration occurs later in G93A-SOD1 mice, with hyperintensity MRI signals detected only at weeks 10-18. Neurodegenerative changes were observed only in the motor nuclei areas of the brainstem; MRI changes indicative of neurodegeneration were not detected in the motor cortex where first motor neurons originate, even at the late disease stage. This longitudinal MRI study establishes unequivocally that, in the experimental murine model of ALS, muscle degeneration occurs before any evidence of neurodegeneration and clinical signs, supporting the postulate that motor neuron disease can initiate from muscle damage and result from retrograde dying-back of the motor neurons. PMID- 21620833 TI - Protection by neuroglobin and cell-penetrating peptide-mediated delivery in vivo: a decade of research. Comment on Cai et al: TAT-mediated delivery of neuroglobin protects against focal cerebral ischemia in mice. Exp Neurol. 2011; 227(1): 224 31. AB - Over the last decade, numerous studies have suggested that neuroglobin is able to protect against the effects of ischemia. However, such results have mostly been based on models using transgenic overexpression or viral delivery. As a therapy, new technology would need to be applied to enable delivery of high concentrations of neuroglobin shortly after the patient suffers the stroke. An approach to deliver proteins in ischemia in vivo in a timely manner is the use of cell penetrating peptides (CPP). CPP have been used in animal models for brain diseases for about a decade as well. In a recent issue of Experimental Neurology, Cai and colleagues test the effect of CPP-coupled neuroglobin in an in vivo stroke model. They find that the fusion protein protects the brain against the effect of ischemia when applied before stroke onset. Here, a concise review of neuroglobin research and the application of CPP peptides in hypoxia and ischemia is provided. PMID- 21620834 TI - Leishmania major: activity of tamoxifen against experimental cutaneous leishmaniasis. AB - Leishmaniasis is a family of diseases caused by protozoan parasites of the genus Leishmania. Various Leishmania species can cause human infection, producing a spectrum of clinical manifestations. The current treatments are unsatisfactory, and in absence of a vaccine, there is an urgent need for effective drugs to replace/supplement those currently in use. Recent studies have shown that the antineoplastic drug, tamoxifen, had direct leishmanicidal effect on several Leishmania species in vitro. Moreover, in vivo testing was carried out on some of the species and showed promising results. The authors have carried out the present work to complement previous published studies by investigating in vivo activity of tamoxifen in an experimental model of cutaneous leishmaniasis (CL) caused by Leishmania major. Groups of infected mice were given tamoxifen, orally, at a dose of 20 mg/kg/day for 15 days. Efficacy was assessed clinically, parasitologically, histopathologically by light and transmission electron microscope (TEM). Results showed that untreated infected mice suffered from autoamputation of the inoculated foot pad. However, those which received tamoxifen showed marked improvement of the cutaneous lesions and reduction of parasite burden. TEM of the cutaneous lesions from infected mice revealed the fine structure of normal Leishmania amastigotes, whereas those from infected mice treated with tamoxifen showed considerable changes. All male mice that received tamoxifen showed scrotal swelling with evident histopathological changes in the testes that could seriously compromise fertility of male mice. In conclusion, although tamoxifen causes significant side effects to the male reproductive system in the mouse model, it could provide an alternative to current agents. Results of this study demonstrated in vivo activity of tamoxifen against Leishmania major, thus, suggesting that tamoxifen is a suitable lead for the synthesis of more effective and less toxic antileishmanial derivatives. PMID- 21620835 TI - Trypanosoma cruzi: desferrioxamine decreases mortality and parasitemia in infected mice through a trypanostatic effect. AB - Desferrioxamine (DFO) is a potent iron chelator that is also known to modulate inflammation and act as an efficient antioxidant under normal conditions and under oxidative stress. Many in vitro and in vivo studies have shown the efficacy of DFO in the treatment of viral, bacterial and protozoan infections. DFO is known to reduce the intensity of Trypanosoma cruzi infections in mice even during a course of therapy that is not effective in maintaining anaemia or low iron levels. To further clarify these findings, we investigated the action of DFO on mouse T. cruzi infection outcomes and the direct impact of DFO on parasites. Infected animals treated with DFO (5 mg/animal/day) for 35 days, beginning 14 days prior to infection, presented lower parasitemia and lower cumulative mortality rate. No significant effect was observed on iron metabolism markers, erythrograms, leukograms or lymphocyte subsets. In the rapid method for testing in vivo T. cruzi susceptibility, DFO also induced lower parasitemia. In regard to its direct impact on parasites, DFO slightly inhibited the growth of amastigotes and trypomastigotes in fibroblast culture. Trypan blue staining showed no effects of DFO on parasite viability, and only minor apoptosis in trypomastigotes was observed. Nevertheless, a clear decrease in parasite mobility was detected. In conclusion, the beneficial actions of DFO on mice T. cruzi infection seem to be independent of host iron metabolism and free of significant haematological side effects. Through direct action on the parasite, DFO has more effective trypanostatic than trypanocidal properties. PMID- 21620836 TI - PI3K-dependent phosphorylation of Fbw7 modulates substrate degradation and activity. AB - The Fbw7 tumor suppressor gene encodes the substrate recognition subunit of the SCF ubiquitin ligase, which targets for degradation a range of oncogenic proteins in a phosphorylation-dependent manner. Substrate phosphorylation is thought to be the main mechanism that ensures timely destruction of Fbw7 substrates. We show here that PI3K dependent phosphorylation of Fbw7 stimulates its ability to ubiquitinate and degrade its substrates. Mutation of the phosphorylation site destabilizes Fbw7 and attenuates degradation of cyclin E and Myc leading to the enhanced expression of a subset of Myc target genes. We suggest that PI3K dependent phosphorylation of Fbw7 controls the balance between turnover of Fbw7 and its substrates to fine-tune their activity. PMID- 21620837 TI - A role for myelin-associated peroxisomes in maintaining paranodal loops and axonal integrity. AB - Demyelinating diseases of the nervous system cause axon loss but the underlying mechanisms are not well understood. Here we show by confocal and electron microscopy that in myelin-forming glia peroxisomes are associated with myelin membranes. When peroxisome biogenesis is experimentally perturbed in Pex5 conditional mouse mutants, myelination by Schwann cells appears initially normal. However, in nerves of older mice paranodal loops become physically unstable and develop swellings filled with vesicles and electron-dense material. This novel model of a demyelinating neuropathy demonstrates that peroxisomes serve an important function in the peripheral myelin compartment, required for long-term axonal integrity. PMID- 21620838 TI - Proton pump inhibitors and safety during pregnancy. PMID- 21620840 TI - Granulocyte-macrophage colony stimulating factor and inflammatory bowel disease: establishing a connection. PMID- 21620842 TI - The cell biology of the unfolded protein response. AB - The unfolded protein response (UPR) is an ensemble of signal transduction pathways that respond to perturbations in the oxidative, pro-folding environment of the endoplasmic reticulum. During the past decade, ongoing research implicated these pathways in maintaining homeostasis of cells and organisms exposed to various stresses. Herein, we highlight recent findings regarding the functional role of the UPR in both normal and pathophysiologic processes. PMID- 21620843 TI - Fruit, vegetables, and folate: cultivating the evidence for cancer prevention. PMID- 21620844 TI - Antibiotics for irritable bowel syndrome: hitting the target, but what is it? PMID- 21620845 TI - Colonoscopy prevents colorectal cancer in both the right and left colon. PMID- 21620846 TI - 'CArG'ing for microRNAs. PMID- 21620847 TI - Will there be an HCV meeting in 2020? Summary of the 17th international meeting on hepatitis C virus and related viruses. PMID- 21620848 TI - Myc, Max, and Mnt: molecular mechanisms of enhancement of cholangiocarcinogenesis by cholestasis. PMID- 21620849 TI - A 13-day, interferon-free regimen for chronic hepatitis C genotype 1 patients: between fear and hope. PMID- 21620850 TI - Recurrent abdominal pain in a 55-year-old woman. Diagnosis: Idiopathic mesenteric phlebosclerosis. PMID- 21620851 TI - Inflammatory response to pyrogens determined by a novel ELISA method using human whole blood. AB - Presence of pyrogens on implants, medical devices, drugs and biological materials compromise on the biosafety and poses a major health hazard in therapeutics. Detection of pyrogenic contamination has so far been done with either in vivo rabbit pyrogen assay or Limulus Amoebocyte Lysate (LAL) methods, each of which having their distinct advantages and disadvantages. An indigenously developed ELISA method quantifying the pro-inflammatory response triggered by pyrogens on human whole blood is demonstrated for its versatility to detect the pyrogenic response to gram-negative, gram-positive bacteria, chemical and biological pyrogens. The method was used to test and quantitate the pyrogen levels in polymeric biomaterials. Unlike the existing pyrogen test procedures, this assay is adapted to detect all pyrogens, besides yielding faster, sensitive and quantifiable data, thereby reduce/replace animal experimentation. The method also provided insight into the possible correlation between variable blood profile among individuals and their role in determining inflammatory response to different pyrogenic stimuli. PMID- 21620852 TI - T cell hybridomas to study MHC-II restricted B-cell receptor-mediated antigen presentation by human B cells. AB - MHC-II antigen presentation by B cells is essential in order for B cells to receive optimal costimulation from helper CD4+ T cells. This process is facilitated and focused through the extremely efficient uptake, processing, and presentation of antigen recognized by an individual B cell's unique B-cell receptor (BCR). The investigation of human B-cell antigen presentation has been limited by the varied specificity of BCR found in the mixed populations of B cells in vivo. As a result, there is no readily available method to measure BCR mediated antigen presentation in this heterogeneous population of B cells. We have overcome this limitation by developing HLA-DR-restricted T-cell lines capable of recognizing a specific antigen taken up via the BCR and presented by the mixed B-cell population through this physiologically relevant mechanism. BCR mediated presentation was enhanced >4 logs compared to presentation by B cells taking up the antigen through nonspecific mechanisms. The studies presented here characterize T-cell hybridoma lines developed for HLA-DRB1*0101+ and HLA DRB1*1501+ B cells, but clones could be generated for other HLA-DR types using the methods described. These hybridomas have potential applications including study of the mechanisms of BCR-mediated enhancement of presentation, determination of adjuvant effects on presentation, and optimization of vaccine antigen preparations. Therefore, these T-cell lines could significantly facilitate the study of BCR-mediated antigen presentation required by T helper cell-dependent vaccines in humans. PMID- 21620853 TI - Characterization of MHC/peptide complexes refolded by a one-step ion-exchange chromatography. AB - The current refolding process of MHC/peptide complexes is low-yield and time consuming, thereby limiting the wide uses of MHC/peptide multimers. Here the heavy chain protein of MHC/peptide complex (H-2K(b)/TRP2(180-188)) was immobilized onto an ion-exchange chromatography column, and the beta2m or TRP2(180-188)-fused beta2m protein, which renatured previously in refolding buffer, was able to pass through the column for the gradient refolding. This strategy refolds, concentrates and purifies MHC/peptide complexes in a single integrated step, achieving a high level of process simplification and automation. Using this on-column refolding method, MHC/peptide complexes could be prepared within 24h with a refolding yield of over 20%. Anti-H-2K(b) mAb staining and flow cytometric analyses revealed that the on-column refolded H-2K(b)/TRP2(180-188) complexes had conformational characteristics similar to the dilution refolded H 2K(b)/TRP2(180-188) complexes and the commercial ones. Furthermore, H 2K(b)/TRP2(180-188) tetramer staining and the enumeration of TRP2(180-188) specific T cells and H-2K(b)-alloreactive T cells confirmed that the H 2K(b)/TRP2(180-188) complexes prepared by on-column refolding or dilution refolding had comparable TCR-binding ability. These data demonstrate a novel, simple and efficient refolding strategy for the generation of MHC class I/peptide complexes. PMID- 21620854 TI - Detection of 3-chlorinated tyrosine residues in human cells by flow cytometry. AB - Hypochlorite is a strong oxidant, generated under pathological conditions, with the potency to introduce chlorine atom into a number of molecules. 3-Chloro- and 3,5-dichlorotyrosine are documented to be generated by this oxidant and their elevated levels were found in many diseases. Thus, we decided to check the possibility of use of FITC-conjugated antibodies for flow cytometric detection of 3-chlorotyrosine residues in human cells (A549, MCF-7, HUVEC-ST) exposed to the action of hypochlorite. Additionally, we compared the effects of chlorohydrins and N-chloroamino acids as chlorine donors. Cell fixation and permeabilization was followed by incubation with rabbit polyclonal anti-3-chlorotyrosine primary antibody and subsequent staining with goat anti-rabbit FITC-labeled secondary antibody. For antibody isotypic control, normal rabbit IgG was employed. Hypochlorite appeared to be the most efficient from the chlorocompounds analyzed in chlorotyrozine generation in all cell lines. Statistically significant increase of fluorescence corresponding to the level of 3-chlorotyrosine residues was found in cells treated with hypochlorite even at non-toxic concentrations (<5MUM). This effect was not observed in cells exposed to the action of chlorinated amino acids or chlorohydrins. The use of anti-3-chlorotyrosine antibodies in conjunction with fluorophore-conjugated secondary antibodies analysis allows for detection of 3-chlorotyrosine residues by flow cytometry in cells treated with low doses of hypochlorite. PMID- 21620855 TI - The use of IgG antibodies in conventional and non-conventional immunodiagnostic tests for early prognosis after treatment of Chagas disease. AB - Treatment success of chronically infected Chagas disease patients is laborious and a positive prognosis often is made only after repetitive serological and/or parasitological examinations with continuous negative results. Recently, we have developed a non-conventional flow-cytometric method in order to detect immunoglobulin G antibodies against live trypomastigote forms of Trypanosoma cruzi and showed its usefulness in the prognosis of treatment success. In the present study, we investigated the performance of flow-cytometric anti-live trypomastigote IgG antibodies (FC-ALTA) and flow-cytometric anti-fixed epimastigote IgG antibodies (FC-AFEA), as well as conventional serological methods, for early monitoring of benznidazole treated Chagas disease patients, e.g. 5years after treatment. The analysis of individual FC-ALTA reactivity along the titration curve before and after treatment, we were able to show, that between 4% and 13% of treated patients under evaluation presented with reduced serological reactivity and segregated from the other patient groups. Similar results were obtained with semi-quantitative, conventional indirect hemagglutination or indirect immunofluorescence. Our data therefore suggest that the combined use of conventional and non-conventional serological methods could provide more suitable cure criteria in early post-therapeutic prognosis of Chagas disease. PMID- 21620856 TI - Effects of ion channels on proliferation in cultured human cardiac fibroblasts. AB - Our previous study demonstrated that multiple ion channels were heterogeneously expressed in human cardiac fibroblasts, including a large-conductance Ca(2+) activated K(+) current (BKCa), a volume-sensitive chloride current (I(Cl.vol)), and voltage-gated sodium currents (I(Na)). The present study was designed to examine the possible involvement of these ion channels in proliferation of cultured human cardiac fibroblasts using approaches of cell proliferation assay, whole-cell patch voltage-clamp, siRNA and Western blot analysis. It was found that the blockade of BKCa with paxilline (1-3MUM) or I(Cl.vol) with 4,4' diisothiocyanatostilbene-2,2'-disulfonic acid disodium (DIDS, 100-200MUM), but not I(Na) with tetrodotoxin (0.1-10MUM), remarkably suppressed proliferation in human cardiac fibroblasts. Knockdown of KCa1.1 or Clcn3 with specific siRNAs significantly reduced BKCa or I(Cl.vol) current, mRNA and channel protein levels, and inhibited growth of human cardiac fibroblasts. Flow cytometry analysis showed accumulation of cardiac fibroblasts at G0/G1 phase and reduced cell number in S phase after inhibition of BKCa or I(Cl.vol) with channel blockers or knock down of the corresponding channels with specific siRNAs; these effects were accompanied by a decreased expression of cyclin D1 and cyclin E. The present results demonstrate the novel information that BKCa and I(Cl.vol) channels, but not I(Na) channels, are involved in the regulation of proliferation in cultured human cardiac fibroblasts by promoting cell cycle progression via modulating cyclin D1 and cyclin E expression. PMID- 21620857 TI - The retinal specific CD147 Ig0 domain: from molecular structure to biological activity. AB - CD147 is a type I transmembrane protein that is involved in inflammatory diseases, cancer progression, and multiple human pathogens utilize CD147 for efficient infection. CD147 expression is so high in several cancers that it is now used as a prognostic marker. The two primary isoforms of CD147 that are related to cancer progression have been identified, differing in their number of immunoglobulin (Ig)-like domains. These include CD147 Ig1-Ig2, which is ubiquitously expressed in most tissues, and CD147 Ig0-Ig1-Ig2, which is retinal specific and implicated in retinoblastoma. However, little is known in regard to the retinal specific CD147 Ig0 domain despite its potential role in retinoblastoma. We present the first crystal structure of the human CD147 Ig0 domain and show that the CD147 Ig0 domain is a crystallographic dimer with an I type domain structure, which maintained in solution. Furthermore, we have utilized our structural data together with mutagenesis to probe the biological activity of CD147-containing proteins, both with and without the CD147 Ig0 domain, within several model cell lines. Our findings reveal that the CD147 Ig0 domain is a potent stimulator of interleukin-6 and suggest that the CD147 Ig0 domain has its own receptor distinct from that of the other CD147 Ig-like domains, CD147 Ig1-Ig2. Finally, we show that the CD147 Ig0 dimer is the functional unit required for activity and can be disrupted by a single point mutation. PMID- 21620858 TI - Structural and thermodynamic signatures of DNA recognition by Mycobacterium tuberculosis DnaA. AB - An essential protein, DnaA, binds to 9-bp DNA sites within the origin of replication oriC. These binding events are prerequisite to forming an enigmatic nucleoprotein scaffold that initiates replication. The number, sequences, positions, and orientations of these short DNA sites, or DnaA boxes, within the oriCs of different bacteria vary considerably. To investigate features of DnaA boxes that are important for binding Mycobacterium tuberculosis DnaA (MtDnaA), we have determined the crystal structures of the DNA binding domain (DBD) of MtDnaA bound to a cognate MtDnaA-box (at 2.0 A resolution) and to a consensus Escherichia coli DnaA-box (at 2.3 A). These structures, complemented by calorimetric equilibrium binding studies of MtDnaA DBD in a series of DnaA-box variants, reveal the main determinants of DNA recognition and establish the [T/C][T/A][G/A]TCCACA sequence as a high-affinity MtDnaA-box. Bioinformatic and calorimetric analyses indicate that DnaA-box sequences in mycobacterial oriCs generally differ from the optimal binding sequence. This sequence variation occurs commonly at the first 2 bp, making an in vivo mycobacterial DnaA-box effectively a 7-mer and not a 9-mer. We demonstrate that the decrease in the affinity of these MtDnaA-box variants for MtDnaA DBD relative to that of the highest-affinity box TTGTCCACA is less than 10-fold. The understanding of DnaA box recognition by MtDnaA and E. coli DnaA enables one to map DnaA-box sequences in the genomes of M. tuberculosis and other eubacteria. PMID- 21620859 TI - Dissecting a bimolecular process of MgATP2- binding to the chaperonin GroEL. AB - Although allosteric transitions of GroEL by MgATP(2)(-) have been widely studied, the initial bimolecular step of MgATP(2-) binding to GroEL remains unclear. Here, we studied the equilibrium and kinetics of MgATP(2)(-) binding to a variant of GroEL, in which Tyr485 was replaced by tryptophan, via isothermal titration calorimetry (ITC) and stopped-flow fluorescence spectroscopy. In the absence of K(+) at 4-5 degrees C, the allosteric transitions and the subsequent ATP hydrolysis by GroEL are halted, and hence, the stopped-flow fluorescence kinetics induced by rapid mixing of MgATP(2)(-) and the GroEL variant solely reflected MgATP(2)(-) binding, which was well represented by bimolecular noncooperative binding with a binding rate constant, k(on), of 9.14*10(4) M(-1) s(-1) and a dissociation rate constant, k(off), of 14.2 s(-1), yielding a binding constant, K(b) (=k(on)/k(off)), of 6.4*10(3) M(-1). We also successfully performed ITC to measure binding isotherms of MgATP(2)(-) to GroEL and obtained a K(b) of 9.5*10(3) M(-1) and a binding stoichiometric number of 6.6. K(b) was thus in good agreement with that obtained by stopped-flow fluorescence. In the presence of 10 50 mM KCl, the fluorescence kinetics consisted of three to four phases (the first fluorescence-increasing phase, followed by one or two exponential fluorescence decreasing phases, and the final slow fluorescence-increasing phase), and comparison of the kinetics in the absence and presence of K(+) clearly demonstrated that the first fluorescence-increasing phase corresponds to bimolecular MgATP(2)(-) binding to GroEL. The temperature dependence of the kinetics indicated that MgATP(2)(-) binding to GroEL was activation-controlled with an activation enthalpy as large as 14-16 kcal mol(-1). PMID- 21620860 TI - Characterization of the interaction of GABARAPL-1 with the LIR motif of NBR1. AB - Selective autophagy requires the specific segregation of targeted proteins into autophagosomes. The selectivity is mediated by autophagy receptors, such as p62 and NBR1, which can bind to autophagic effector proteins (Atg8 in yeast, MAP1LC3 protein family in mammals) anchored in the membrane of autophagosomes. Recognition of autophagy receptors by autophagy effectors takes place through an LC3 interaction region (LIR). The canonical LIR motif consists of a WXXL sequence, N-terminally preceded by negatively charged residues. The LIR motif of NBR1 presents differences to this classical LIR motif with a tyrosine residue and an isoleucine residue substituting the tryptophan residue and the leucine residue, respectively. We have determined the structure of the GABARAPL-1/NBR1 LIR complex and studied the influence of the different residues belonging to the LIR motif for the interaction with several mammalian autophagy modifiers (LC3B and GABARAPL-1). Our results indicate that the presence of a tryptophan residue in the LIR motif increases the binding affinity. Substitution by other aromatic amino acids or increasing the number of negatively charged residues at the N terminus of the LIR motif, however, has little effect on the binding affinity due to enthalpy-entropy compensation. This indicates that different LIRs can interact with autophagy modifiers with unique binding properties. PMID- 21620861 TI - Collective motion from local attraction. AB - Many animal groups, for example schools of fish or flocks of birds, exhibit complex dynamic patterns while moving cohesively in the same direction. These flocking patterns have been studied using self-propelled particle models, most of which assume that collective motion arises from individuals aligning with their neighbours. Here, we propose a self-propelled particle model in which the only social force between individuals is attraction. We show that this model generates three different phases: swarms, undirected mills and moving aligned groups. By studying our model in the zero noise limit, we show how these phases depend on the relative strength of attraction and individual inertia. Moreover, by restricting the field of vision of the individuals and increasing the degree of noise in the system, we find that the groups generate both directed mills and three dynamically moving, 'rotating chain' structures. A rich diversity of patterns is generated by social attraction alone, which may provide insight into the dynamics of natural flocks. PMID- 21620862 TI - Positive effects of multiple gene control on the spread of altruism by group selection. AB - The origin of altruistic behavior has long been a challenge for students of evolutionary biology. The populations with altruistic individuals do better than those without altruists; however, the altruists within a population do worse than the non-altruists and their prevalence in the population decreases due to individual selection. Under certain conditions, the strength of group selection, i.e., the selection driven by competition between populations, can surpass the strength of individual selection; however, such conditions seem to be relatively strict and probably do not hold in many natural systems where the altruistic behavior was observed. It was suggested recently that chances for altruistic behavior to spread highly increase when it is controlled not by a single gene but by multiple independent genes substitutable in their effects on the phenotype of the individual. Here we confirm the original verbal model by numerical modeling of the spread of altruistic/selfish alleles in a metapopulation consisting of partly isolated groups of organisms (demes) interconnected by migration. We have shown that altruistic behavior coded by multiple substitutable genes can stably coexist with selfish behavior, even under relatively high mutation and migration rates, i.e., under such conditions where altruistic behavior coded by a single gene is quickly outcompeted in a metapopulation. PMID- 21620863 TI - Helix-helix interfaces and ligand binding. AB - Helix-helix parallel interfaces can be characterized by certain combinations of amino acids, which repeatedly occur at core positions a and d (leucine zipper nomenclature) in homologous and nonhomologous proteins and influence interhelical angles. Applied for the prediction of interhelical angles in glutathione S transferase, intracellular chloride channel and annexin molecules from various sources, correct results were achieved in 58 out of 62 proteins. Interhelical angles are found to correlate with the conformation of the glutathione S transferase ligands glutathione, s-hexylglutathione, glutathione sulfonic acid, and glutathione-s-dinitrobenzene. PMID- 21620864 TI - Hierarchical organization of noise generates spontaneous signal in Paramecium cell. AB - In many cellular processes, spontaneous activities are often the basis for their functioning. Paramecium cells change their swimming direction under a homogeneous environment, which is induced by a spontaneous signal generation in the membrane electric potential. For such a spontaneous activity, a theoretical model has been proposed by Oosawa (2007) [Biosystems 88, 191-201.], in which intracellular noise is hierarchically organized from thermal fluctuations to spike-like large fluctuations, which induces a signal to change spontaneously the swimming direction. Our analysis of the model shows that the system is a kind of excitable media, in which a spike is induced by a stochastic fluctuation. We show conditions of channels properties to have a spike train. PMID- 21620865 TI - Does cutting herbicide rates threaten the sustainability of weed management in cropping systems? AB - Evolution of herbicide resistance in weeds is a growing problem across the world, and it has been suggested that low herbicide rates may be contributing to this problem. An individual-based simulation model that represents weed population dynamics and the evolution of polygenic herbicide resistance was constructed and used to investigate whether using lower herbicide rates or standard rates at reduced efficacy could reduce the sustainability of cropping systems by causing faster increases in weed population density as herbicide resistance develops. A number of different possible genetic bases for resistance were considered, including monogenic resistance and polygenic resistance conferred by several genes. The results show that cutting herbicide rates does not affect the rate at which weed densities reach critical levels when resistance is conferred exclusively by a single dominant gene. In some polygenic situations, cutting herbicide rates substantially reduces sustainability, due to a combination of faster increase in resistance gene frequency and reduced kill rates in all genotypes, while in other polygenic situations the effect is small. Differences in sustainability depend on combined strength of the resistance genes, variability in phenotypic susceptibility and rate delivered, level of control due to alternative measures, and degree of genetic dominance and epistasis. In the situation where resistance can be conferred by both a single dominant major gene or a number of co-dominant minor genes in combination, the difference made by low rates depends on the relative initial frequency of the major and minor genes. These results show that careful consideration of herbicide rate and understanding the genetic basis of resistance are important aspects of weed management. PMID- 21620866 TI - Exercise training has beneficial anti-atrophy effects by inhibiting oxidative stress-induced MuRF1 upregulation in rats with diabetes. AB - AIMS: MuRF1 E3 ubiquitin ligase has been identified as a mediator of skeletal muscle wasting in various skeletal muscle atrophy models, and its expression is upregulated by oxidative stress. Exercise training could decrease oxidative stress and restore the atrophied skeletal muscle. Here, our aim was to investigate whether exercise training has any effect on MuRF1 expression in rats with diabetes. MAIN METHODS: Rats with streptozotocin-induced diabetes were subjected to exercise training, after which oxidative stress was determined, and MuRF1 expression was analyzed by immunohistochemistry, real-time RT-PCR and Western blot analysis. In addition, we analyzed C2C12 myotubes in an in vitro model to examine the effects of oxidative stress on the protein levels of MuRF1 and myosin heavy chain (MHC). KEY FINDINGS: While oxidative stress and MuRF1 expression were increased in rats with diabetes, exercise training diminished the skeletal muscle wasting in diabetic rats by decreasing oxidative stress and inhibiting MuRF1 expression at both the mRNA and protein levels. In addition, oxidative stress-induced MuRF1 upregulation promoted proteasome dependent degradation of the myosin heavy chain (MHC) in C2C12 myotubes. SIGNIFICANCE: Our study provides the first evidence that the beneficial anti-atrophy effects of exercise training on diabetes might be mediated by inhibiting oxidative stress induced MuRF1 upregulation and preventing MuRF1-mediated degradation of MHC. PMID- 21620867 TI - Protective effect of telmisartan against cadmium-induced nephrotoxicity in mice. AB - AIMS: To investigate the nephroprotective effect of telmisartan, the angiotensin II receptor antagonist, against renal injury induced by cadmium in mice. MAIN METHODS: Mice received cadmium chloride at a dose of 1.2mg Cd/kg/day, s.c., for nine weeks. Telmisartan treatment (1mg/kg/day, orally) was started one week before cadmium administration and continued for ten weeks. KEY FINDINGS: Telmisartan significantly reduced blood urea nitrogen (BUN) and serum creatinine levels which were increased by cadmium. Also, telmisartan significantly suppressed lipid peroxidation, compensated deficits in the antioxidant defenses [reduced glutathione (GSH) level and catalase activity], decreased the elevations of tumor necrosis factor-alpha (TNF-alpha), nitric oxide (NO) and cadmium ion concentration, and attenuated the reductions of selenium and zinc ions in renal tissue resulted from cadmium administration. Histopathological examination revealed that cadmium-induced renal tissue damage was ameliorated by telmisartan treatment. Immunohistochemical analysis revealed that telmisartan significantly decreased the cadmium-induced overexpression of inducible nitric oxide synthase (iNOS), nuclear factor-kappaB (NF-kappaB), Fas ligand (FasL) and caspase-3 in renal tissue. SIGNIFICANCE: Telmisartan, through its antioxidant and anti inflammatory actions, effectively prevented cadmium nephrotoxicity in mice. Hence, telmisartan represents a potential candidate to protect the kidney from the detrimental effect of cadmium toxicity. PMID- 21620868 TI - Cerebral ischemia aggravates cognitive impairment in a rat model of Alzheimer's disease. AB - AIM: Autopsy evidence suggests that the presence of both Alzheimer(')s disease (AD) and cerebral infarction pathology is associated with more severe cognitive impairment than that produced by AD pathology alone. This study aims to investigate the effect of cerebral ischemia on cognitive function in rats with AD constructed by hippocampal injection and to determine its underlying mechanism, which is proposed to be of significance to the treatment of AD. MAIN METHODS: AD was modeled by injection of aggregated Abeta(1-40), either alone or followed by hippocampal endothelin-1 injection to mimic cerebral ischemia in hippocampus, into the right dentate gyrus (DG) of rats. The Morris water maze was used to evaluate cognitive function. Abeta deposition, neuronal loss and phosphorylated tau expression in hippocampus were examined by Congo red staining, Nissl's staining and immunohistochemistry, respectively. Reactive astrocytes, IL-1beta and TNF-alpha expressions were measured by immunohistochemistry, in situ hybridization and reverse transcription-polymerase chain reaction. KEY FINDINGS: Compared with rats treated with either Abeta or endothelin alone, rats treated with both Abeta and endothelin showed more aggravated cognitive impairment and more Abeta deposits, neuron loss, phosphorylated tau expression, reactive astrocytes, IL-1beta and TNF-alpha expressions in hippocampus. SIGNIFICANCE: Hippocampal ischemia aggravates cognitive impairment of AD rats by increasing Abeta deposits, neuron loss and tau phosphorylation in hippocampus. The enhanced inflammatory response may be responsible for cerebral ischemia-induced aggravation of cognitive impairment in AD rats. Based on these findings, prevention and treatment of cerebral ischemia may improve clinical symptoms of AD and suppress the progression of AD. PMID- 21620869 TI - Thiosemicarbazone derivate protects from AAPH and Cu2+ -induced LDL oxidation. AB - AIMS: Several lines of evidence support the hypotheses that the oxidation of low density lipoprotein (LDL) may play a crucial role in the initiation and progression of atherosclerosis. Oxidative stress is one of the causes of the overproduction of reactive species that increase the formation of oxidized LDL. Thiosemicarbazones are compounds used in anticancer, antiviral and antifungal therapy; however, its redox activity has been controversial. Thus, we tested, in vitro, a possible antioxidant activity of a thiosemicarbazone derivate, the isatin-3-N(4)-benzilthiosemicarbazone (IBTC). MAIN METHODS: We measured the conjugated diene formation in serum and LDL as well as the loss of tryptophan fluorescence in LDL induced by two oxidant agents, 2,2-azobis(2-amidinopropane dihydrochloride) (AAPH) and Cu(2+). Thiobarbituric acid reactive substances (TBARS) formation in LDL and in different rat tissues was also assessed. The toxicity of IBTC was measured using aortic slices viability assay. KEY FINDINGS: Our results show that IBTC significantly reduced the AAPH and Cu(2+)-induced formation of conjugated dienes, increased in a dose-dependent manner the lag phase and the t(1/2) of tryptophan fluorescence, and reduced the TBARS formation in LDL, plasma and rat tissues, showing no toxicity to aortic slices. SIGNIFICANCE: These results indicate that IBTC is a good antioxidant and a promising antiatherogenic agent for further studies in vivo. PMID- 21620870 TI - Essential oils components as a new path to understand ion channel molecular pharmacology. AB - The discovery and development of new drugs targeting voltage-gated ion channels are important for treating a variety of medical conditions and diseases. Ion channels are molecular nanostructures expressed ubiquitously throughout the whole body, and are involved in many basic physiological processes. Over the years, natural products have proven useful in the pharmacological assessment of ion channel structure and function, while also contributing to the identification of lead molecules for drug development. Essential oils are complex chemical mixtures isolated from plants which may possess a large spectrum of biological activities most of them of clinical interest. Among their bioactive constituents, terpenes are small to medium-sized components and belong to different chemical groups. Various reports have drawn our attention to the fact that terpenes are novel compounds targeting voltage-gated ion channels. The purpose of this review is to provide a focused discussion on the molecular interaction between monoterpenes and phenylpropenes with voltage-gated ion channels in different biological scenarios. PMID- 21620871 TI - Combination therapy with bone marrow stromal cells and FK506 enhanced amelioration of ischemic brain damage in rats. AB - AIMS: Transplantation of bone marrow stromal cells (MSCs) has been shown to ameliorate ischemic brain injury in animals. In the present study, we investigated whether the transplantation of MSCs combined with FK506, a clinically used immunosuppressant, enhanced neuroprotective effects in rat experimental stroke. MAIN METHODS: Male Sprague-Dawley rats underwent transient 90 min middle cerebral artery occlusion (MCAO). Two or 6h after ischemia onset, the rats were randomly assigned to receive intravenous administration of MSCs plus FK506, MSCs alone, FK506 alone, or vehicle. Infarct volume, and neurological and immunohistological assessments were performed to examine the effects of these therapies. KEY FINDINGS: In 2-hour post-ischemia treatment groups, significant improvement of infarct volume and neurological scores were observed 1 day after combination therapy compared with monotherapy, and this neuroprotection continued for 7 days. Combination therapy significantly reduced the number of TUNEL positive apoptotic cells, increased Bcl-2 expression, decreased Bax expression, and suppressed neutrophil infiltration and microglia/macrophage activation compared to monotherapy. In 6-hour post-ischemia treatment groups, a significant reduction of infarct volume, edema index, and neurological score was observed only in the combination therapy group. Moreover, the number of engrafted MSCs on day 7 with combination therapy was significantly higher than with MSCs alone. SIGNIFICANCE: Combination therapy using FK506 enhanced the anti-apoptotic and anti-inflammatory effects of MSCs and increased the survival of transplanted cells, leading to expansion of the therapeutic time window for MSCs. PMID- 21620872 TI - The renin-angiotensin system is modulated by swimming training depending on the age of spontaneously hypertensive rats. AB - AIM: To investigate the effects of swimming training on the renin-angiotensin system (RAS) during the development of hypertensive disease. MAIN METHODS: Male spontaneously hypertensive rats (SHR) were randomized into: sedentary young (SY), trained young (TY), sedentary adult (SA), and trained adult (TA) groups. Swimming was performed 5 times/wk/8wks. KEY FINDINGS: Trained young and adult rats showed both decreased systolic and mean blood pressure, and bradycardia after the training protocol. The left ventricular hypertrophy (LVH) was observed only in the TA group (12.7%), but there was no increase on the collagen volume fraction. Regarding the components of the RAS, TY showed lower activity and gene expression of angiotensinogen (AGT) compared to SY. The TA group showed lower activity of circulatory RAS components, such as decreased serum ACE activity and plasma renin activity compared to SA. However, depending on the age, although there were marked differences in the modulation of the RAS by training, both trained groups showed a reduction in circulating angiotensin II levels which may explain the lower blood pressure in both groups after swimming training. SIGNIFICANCE: Swimming training regulates the RAS differently in adult and young SHR rats. Decreased local cardiac RAS may have prevented the LVH exercise-induced in the TY group. Both groups decreased serum angiotensin II content, which may, at least in part, contribute to the lowering blood pressure effect of exercise training. PMID- 21620873 TI - Single early prenatal lipopolysaccharide exposure prevents subsequent airway inflammation response in an experimental model of asthma. AB - AIMS: There has been emerging interest in the prenatal determinants of respiratory disease. In utero factors have been reported to play a role in airway development, inflammation, and remodeling. Specifically, prenatal exposure to endotoxins might regulate tolerance to allergens later in life. The present study investigated whether prenatal lipopolysaccharide (LPS) administration alters subsequent offspring allergen-induced inflammatory response in adult rats. MAIN METHODS: Pregnant Wistar rats were treated with LPS (100 MUg/kg, i.p.) on gestation day 9.5 and their ovariectomized female offspring were sensitized and challenged with OVA later in adulthood. The bronchoalveolar lavage (BAL) fluid, peripheral blood, bone marrow leukocytes and passive cutaneous anaphylaxis were evaluated in these 75-day-old pups. KEY FINDINGS: OVA sensitized pups of NaCl treated rats showed an increase of leucocytes in BAL after OVA challenge. This increase was attenuated, when mothers were exposed to a single LPS injection early in pregnancy. Thus, LPS prenatal treatment resulted in (1) lower increased total and differential (macrophages, neutrophils, eosinophils and lymphocytes) BAL cellularity count; (2) increased number of total, mononuclear and polymorphonuclear cells in the peripheral blood; and (3) no differences in bone marrow cellularity or passive cutaneous anaphylaxis. SIGNIFICANCE: In conclusion, female pups treated prenatally with LPS presented an attenuated response to experimentally-induced asthma. We observed reduced immune cell migration from peripheral blood to the lungs, with no effect on the production of bone marrow cells or antibodies. It was suggested that inflammatory events such as exposure to LPS in early fetal life can attenuate allergic inflammation in the lung, which is a common symptom in asthma. PMID- 21620874 TI - Role of high-fat diet in regulation of gene expression of drug metabolizing enzymes and transporters. AB - AIM: Our aim is to investigate the molecular mechanism of regulation of gene expression of drug metabolizing enzymes (DMEs) and transporters in diet-induced obesity. MAIN METHODS: Adult male CD1 mice were fed diets containing 60% kcal fat (HFD) or 10% kcal fat (LFD) for 14 weeks. RNA levels of hepatic DMEs, transporters and their regulatory nuclear receptors (NRs) were analyzed by real time PCR. Activation of cell-signaling components (JNK and NF-kappaBeta) and pro inflammatory cytokines (IL-1beta, IL-6 and TNFalpha) were measured in the liver. Finally, the pharmacodynamics of drugs metabolized by DMEs was measured to determine the clinical relevance of our findings. KEY FINDINGS: RNA levels of the hepatic phase I (Cyp3a11, Cyp2b10, Cyp2a4) and phase II (Ugt1a1, Sult1a1, Sultn) enzymes were reduced ~30-60% in HFD compared to LFD mice. RNA levels of Cyp2e1, Cyp1a2 and the drug transporters, multidrug resistance proteins, (Mrp)2, Mrp3 and multidrug resistant gene (Mdr)1b were unaltered in HFD mice. Gene expression of the NRs, PXR and CAR and nuclear protein levels of RXRalpha was reduced in HFD mice. Cytokines, JNK and NF-kappaBeta were induced in HFD mice. Thus reduction in hepatic gene expression in obesity may be modulated by cross-talk between NRs and inflammation-induced cell-signaling. Sleep time of Midazolam (Cyp3a substrate) was prolonged in HFD mice, while Zoxazolamine (Cyp1a2 and Cyp2e1 substrate) induced sleep time was unaltered. SIGNIFICANCE: This study demonstrates that gene specific reductions in DMEs can affect specific drugs metabolized by these enzymes, thus providing a rationale to monitor the effectiveness of drug therapy in obese individuals. PMID- 21620875 TI - Cigarette smoke extract activates human bronchial epithelial cells affecting non neuronal cholinergic system signalling in vitro. AB - AIMS: Acetylcholine (ACh) is synthesized by Choline Acetyl-Transferase (ChAT) that exerts its physiological effects in airway epithelial cells via muscarinic receptor (MR) activation. We evaluate the effect of ACh stimulation on human bronchial epithelial cells (16-HBE) and test whether cigarette smoke extract (CSE) can modify the basal cellular response to ACh affecting the non-neuronal cholinergic system signalling. MAIN METHODS: ACh stimulated 16-HBE were tested for ACh-binding, Leukotriene B(4) (LTB(4)) release and ERK1/2 and NFkB pathway activation. Additionally, we investigated all the aforementioned parameters as well as ChAT and MR proteins and mRNA expression and endogenous ACh production in CSE-treated 16-HBE. KEY FINDINGS: We showed that ACh induced in 16-HBE, in a concentration-dependent manner, LTB(4) release via the activation of ERK1/2 and NFkB pathways. The addition of Tiotropium (Spiriva(r)), Gallamine, Telenzepine and 4-DAMP (muscarinic receptor antagonists), as well as of PD 098059 (MAPKK inhibitor) and BAY117082 (inhibitor of IkBalpha phosphorilation), down-regulated the ACh-induced effects. Additionally, CSE treatment of 16-HBE increased the binding of ACh, and shifted the LTB4 release from the concentration ACh 1MUM to 10nM. Finally, we observed that the treatment of 16-HBE with CSE increased the expression of ChAT, M(2) and M(3) and of endogenous ACh production in 16-HBE. Tiotropium regulated the LTB4 release and ACh production in CSE treated 16-HBE. SIGNIFICANCE: CSE increases the pro-inflammatory activity of human bronchial epithelial cells, and promotes the cellular response to lower concentrations of ACh, by affecting the expression of ChAT and MRs. Tiotropium might prevent pro inflammatory events generated by ACh together with CSE. PMID- 21620877 TI - Alzheimer's disease and memory-monitoring impairment: Alzheimer's patients show a monitoring deficit that is greater than their accuracy deficit. AB - We assessed the ability of two groups of patients with mild Alzheimer's disease (AD) and two groups of older adults to monitor the likely accuracy of recognition judgments and source identification judgments about who spoke something earlier. Alzheimer's patients showed worse performance on both memory judgments and were less able to monitor with confidence ratings the likely accuracy of both kinds of memory judgments, as compared to a group of older adults who experienced the identical study and test conditions. Critically, however, when memory performance was made comparable between the AD patients and the older adults (e.g., by giving AD patients extra exposures to the study materials), AD patients were still greatly impaired at monitoring the likely accuracy of their recognition and source judgments. This result indicates that the monitoring impairment in AD patients is actually worse than their memory impairment, as otherwise there would have been no differences between the two groups in monitoring performance when there were no differences in accuracy. We discuss the brain correlates of this memory-monitoring deficit and also propose a Remembrance-Evaluation model of memory-monitoring. PMID- 21620876 TI - Metabotropic glutamate receptors as therapeutic targets for schizophrenia. AB - Treatment options for schizophrenia that address all symptom categories (positive, negative, and cognitive) are lacking in current therapies for this disorder. Compounds targeting the metabotropic glutamate (mGlu) receptors hold promise as a more comprehensive therapeutic alternative to typical and atypical antipsychotics and may avoid the occurrence of extrapyramidal side effects that accompany these treatments. Activation of the group II mGlu receptors (mGlu(2) and mGlu(3)) and the group I mGlu(5) are hypothesized to normalize the disruption of thalamocortical glutamatergic circuitry that results in abnormal glutamaterigic signaling in the prefrontal cortex (PFC). Agonists of mGlu(2) and mGlu(3) have demonstrated efficacy for the positive symptom group in both animal models and clinical trials with mGlu(2) being the subtype most likely responsible for the therapeutic effect. Limitations in the chemical space tolerated by the orthosteric site of the mGlu receptors has led to the pursuit of compounds that potentiate the receptor's response to glutamate by acting at less highly conserved allosteric sites. Several series of selective positive allosteric modulators (PAMs) for mGlu(2) and mGlu(5) have demonstrated efficacy in animal models used for the evaluation of antipsychotic agents. In addition, evidence from animal studies indicates that mGlu(5) PAMs hold promise for the treatment of cognitive deficits that occur in schizophrenia. Hopefully, further optimization of allosteric modulators of mGlu receptors will yield clinical candidates that will allow full evaluation of the potential efficacy of these compounds in the treatment of multiple symptom domains in schizophrenia patients in the near future. PMID- 21620878 TI - Weight bearing evaluation in inflammatory, neuropathic and cancer chronic pain in freely moving rats. AB - Preclinical pain assessment remains a key step for the development of new and potent painkillers. Significant progress in pain evaluation has been achieved with the development of non-reflexive tools. Seeking efficient and clinically relevant devices for pain-related quality of life assessment, we evaluated a new Dynamic Weight Bearing (DWB) device based on pressure captors in three different preclinical chronic pain models. Inflammatory (CFA), neuropathic (CCI) and bone cancer pain (femoral tumor) models were evaluated in Sprague Dawley rats for mechanical allodynia using dynamic von Frey for pain-related behaviors and DWB for discomfort. We observed similar impairment patterns in all of the models for both von Frey (allodynia) and DWB (weight balance) during the complete observation period, starting at day 3 in CCI- and CFA-affected limbs and at day 14 in bone cancer-afflicted rats, indicating that the DWB could be a useful tool for supporting pain assessment. Interestingly, we demonstrated that the main compensation, when animals experienced pain, was seen in the forepaws, ranging from 46% to 69% of increased load compared to normal. Other pain-related coping behaviors were also measured, such as the time spent on each paw and the contact surface. Our results revealed that CFA, CCI and cancerous rats decreased the use of their ipsilateral hind paws by 30% and showed a 50% reduction in paw surface pressed against the floor. In conclusion, this new device improves methods for preclinical evaluation of discomfort and quality of life proxies and could be helpful in screening putative analgesics. PMID- 21620879 TI - Physical and related sensory properties of a swallowable bolus. AB - Rheology and water content properties of cereal boluses collected just before swallowing were investigated. No specific physical markers for swallowing were found between subjects. Each subject had his own mastication strategy leading to food boluses with different rheological and water content properties. However, for most of the subjects, similar physical properties were found for food boluses obtained from consumption of different cereals. Results showed that the food boluses from different cereals exhibited gel-like properties being in a range from 14.1 kPa to 21.2 kPa (G'(1 Hz, 0.4%)), when swallowed. The food boluses had a static yield stress varying from 1.3 kPa to 4.3 kPa. Another interesting finding was that the water content of food boluses might be an important marker for swallowing since it was similar for different cereal food boluses (around 50%). This physical property might drive the fluid sensory perception, which could also be a sensory swallowing threshold. PMID- 21620880 TI - Promoting children's healthy eating in obesogenic environments: Lessons learned from the rat. AB - Current statistics on children's eating patterns and obesity rates are consistent with the idea that genetic taste predispositions, traditional feeding practices, and the obesogenic environment combine to increase the likelihood of unhealthy outcomes in many individuals. In this paper, we focus on one particular level of analysis through which this unhealthy combination of factors may begin to be disassembled: children's learning about food and flavors. Much of the research on children's learning about food and flavors has been inspired by the animal literature, which has a long history of carefully controlled studies elucidating the mechanisms through which rats and other animals learn to prefer and avoid foods and flavors. This literature provides many clues as to the processes by which learning paradigms may be used to encourage the intake of healthy foods, altering the implicit learning of obesogenic eating patterns that is likely to occur without intervention in the current environment. Overall, the implications of the literature are that children should be repeatedly exposed to a variety of flavors early in life, and that new flavors should be paired with already-liked flavors and positive contexts. This message is consistent with recent research results from our laboratory, showing that familiarization and associative learning paradigms may be used to increase young children's acceptance of, preference for, and intake of previously-unfamiliar, healthy foods. PMID- 21620881 TI - Flu-like symptoms following botulinum toxin therapy. AB - This paper reviews the relevant literature on flu-like symptoms (FLS) in patients treated with botulinum toxin (BoNT) type A and B. We conducted a systematic search in PubMed and Medline of publications on BoNT and FLS. FLS were defined as the presence of symptoms of an upper respiratory tract infection associated with fever, general malaise or fatigue, within one month after BoNT injections and not secondary to any other identified condition. FLS have been reported in between 1.7 and 20% of patients treated with various preparations of BoNT/A. Most patients have a mild to moderate symptoms lasting less than 2 weeks, but 66 serious AEs related to FLS were reported to the Food and Drug Administration between 1989 and 2003. No significant differences in the frequency of FLS have emerged when different BoNT/A commercial products were compared, although no well designed head-to-head comparison trials focusing on FLS have been published. For BoNT/B preparations, FLS have been reported in 5-55% of cases. We concluded that the frequency of FLS in patients treated with BoNT/A and BoNT/B varies widely between studies and no clear risk factors have been identified for this complication. We suggest potential treatments like analgesics/antipyretics or switching to less antigenic preparations in case of persistent symptoms. PMID- 21620882 TI - Chicoric acid regulates behavioral and biochemical alterations induced by chronic stress in experimental Swiss albino mice. AB - The present study was taken up to see the effect of chicoric acid (CA) on behavioral and biochemical alterations induced by chronic restraint stress in experimental Swiss albino mice. CA at 1mg/kg dose level exhibited considerable antidepressant activity as shown by significant decrease in immobility period in the Porsolt's swim stress-induced behavioral despair test and escape failures in Learned "helplessness test". The antidepressant activity shown by CA can be attributed to its modulating effect on nor-adrenaline (NA), dopamine (DA) and 5- hydroxy tryptamine (5-HT) as shown by their quantification in CA treated chronically stressed mice. Further, a significant antioxidant effect was exhibited by CA as shown by estimation of lipid peroxidation, glutathione (GSH) and glycogen in liver of chronically stressed mice. It also normalized altered values of serum glucose, triglycerides, aspartate aminotransferase (AST) alanine aminotransferase (ALT) and alkaline phosphatase (ALP) in a dose dependent manner. The stress busting potential of CA was further confirmed by its regulating effect on raised plasma corticosterone levels and significant attenuation of the depleted ascorbic acid, cholesterol and corticosterone levels in adrenal glands. Thus, our results suggest that CA possesses considerable stress busting potential, and that anti-oxidation may be one of the mechanisms underlying its antistress action. PMID- 21620883 TI - Aged monkeys as a partial model for Parkinson's disease. AB - Parkinson's Disease (PD) and the natural aging process share a number of biochemical mechanisms, including reduced function of dopaminergic systems. The present study aims to determine the extent that motor and behavioral changes in aged monkeys resemble parkinsonism induced by the neurotoxin 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine. The behavioral and physiological changes in PD are believed to result largely from selective depletion of dopamine in the nigrostriatal system. In the present study, ten aged female monkeys were compared with three groups: 9 untreated young adult female monkeys, 10 young adult male monkeys and 13 older male monkeys that had been exposed to MPTP. Trained observers, blind as to age and drug condition and without knowledge of the hypotheses, scored the monkeys using the Parkinson's factor score (Parkscore), which has been validated by a high correlation with post mortem striatal dopamine (DA) concentrations. The aged animals had higher scores on the Parkscore compared with the young adults, with most of its component behavioral items showing significance (tremor, Eating Problems, Delayed initiation of movement, and Poverty of Movement). L-Dopa and DA-agonists did not clearly reverse the principal measure of parkinsonism. DA concentrations post mortem were 63% lower in 3 aged monkeys in the ventral putamen compared with 4 young adults, with greater reductions in putamen than in caudate (45%). We conclude that aged monkeys, unexposed to MPTP, show a similar profile of parkinsonism to that seen after the neurotoxin exposure to MPTP in young adult monkeys. The pattern of greater DA depletion in putamen than in caudate in aged monkeys is the same as in human Parkinson's disease and contrasts with the greater depletion in caudate seen after MPTP. Aged monkeys of this species reflect many facets of Parkinson's disease, but like older humans do not improve with standard dopamine replacement pharmacotherapies. PMID- 21620884 TI - Secondary structure of chorion proteins of the Lepidoptera Pericallia ricini and Ariadne merione by ATR FT-IR and micro-Raman spectroscopy. AB - The gross morphological features of the eggs and eggshells (chorions) of two Lepidoptera species, Pericallia ricini and Ariadne merione were revealed for the first time by scanning and transmission electron microscopy. These two insect pests are extremely serious threats for many crops, mainly in India, but also in several other regions of the world. Micro-Raman and ATR FT-IR spectroscopy were also applied to study in detail the secondary structure of the eggshell (chorion) proteins of these Lepidoptera species. Both techniques indicate that the two species have nearly identical conformations of their chorion proteins with abundant antiparallel beta-pleated sheet. These results are in support of our previous findings that the helicoidal architecture of the proteinaceous chorion of Lepidoptera and fishes is dictated by a common molecular denominator, the antiparallel beta-pleated sheet secondary structure. PMID- 21620885 TI - repABC-based replication systems of Rhizobium leguminosarum bv. trifolii TA1 plasmids: incompatibility and evolutionary analyses. AB - Soil bacteria of the genus Rhizobium possess complex genomes consisting of a chromosome and in addition, often, multiple extrachromosomal replicons, which are usually equipped with repABC genes that control their replication and partition. The replication regions of four plasmids of Rhizobium leguminosarum bv. trifolii TA1 (RtTA1) were identified and characterized. They all contained a complete set of repABC genes. The structural diversity of the rep regions of RtTA1 plasmids was demonstrated for parS and incalpha elements, and this was especially apparent in the case of symbiotic plasmid (pSym). Incompatibility assays with recombinant constructs containing parS or incalpha demonstrated that RtTA1 plasmids belong to different incompatibility groups. Horizontal acquisition was plausibly the main contributor to the origin of RtTA1 plasmids and pSym is probably the newest plasmid of this strain. Phylogenetic and incompatibility analyses of repABC regions of three closely related strains: RtTA1, R. leguminosarum bv. viciae 3841 and Rhizobium etli CFN42, provided data on coexistence of their replicons in a common genomic framework. PMID- 21620886 TI - Concerns about cultural neurosciences: a critical analysis. AB - Ten years ago, neuroscientists began to study cultural phenomena by using functional MRI. Since then the number of publications in this field, termed cultural neuroscience (CN), has tremendously increased. In these studies, particular concepts of culture are implied, but rarely explicitly discussed. We argue that it is necessary to make these concepts a topic of debate in order to unravel the foundations of CN. From 40 fMRI studies we extracted two strands of reasoning: models investigating universal mechanisms for the formation of cultural groups and habits and, models assessing differences in characteristics among cultural groups. Both strands simplify culture as an inflexible set of traits and specificities. We question this rigid understanding of culture and highlight its hidden evaluative nature. PMID- 21620887 TI - beta-Hexachlorocyclohexane levels in serum and risk of Parkinson's disease. AB - Pesticide exposure has been implicated as an environmental risk factor for the development of Parkinson's disease (PD). However, few studies have identified specific pesticides. Previously, we identified elevated serum levels of the organochlorine pesticide beta-hexachlorocyclohexane (beta-HCH) in PD patients from a small clinical sample. Here, we conducted a case-control study to confirm the association between beta-HCH and PD in a larger sample size (n=283) with serum samples of PD patients and controls obtained from UT Southwestern Medical Center and Emory University. Samples were obtained from two discrete periods at both sites, 2001-2003 and 2006-2008, and were analyzed for beta-HCH levels. Adjusted odds ratios (ORs) for PD were estimated using logistic regression and generalized estimating equations. The mean serum beta-HCH level across all cohorts in this study was 22.3 ng/mg cholesterol (range: 0-376.7), and the levels were significantly higher and samples collected in 2001-2003 vs. 2006-2008. After controlling for age and gender, the OR for increased risk of PD for every 1 ng/mg increase in serum beta-HCH ranged from 1.02 to 1.12 across the four different cohorts, and 1.03 (95% CI: 1.00-1.07, p value=0.031) in the pooled analysis. Furthermore, the OR for increased risk of PD of subjects having serum beta-HCH levels above the inter-quartile range of 39.08 ng/mg cholesterol was 2.85 (95% CI: 1.8, 4.48; p value<0.001). These data are consistent with environmental decreases in beta-HCH levels between 2001 and 2008, but they indicate that elevated levels of serum beta-HCH are still associated with heightened risk for PD. PMID- 21620889 TI - The contribution of the Drosophila model to lipid droplet research. AB - Intracellular lipid droplets have long been misconceived as evolutionarily conserved but functionally frugal components of cellular metabolism. An ever growing repertoire of functions has elevated lipid droplets to fully-fledged cellular organelles. Insights into the multifariousness of these organelles have been obtained from a range of model systems now employed for lipid droplet research including the fruit fly, Drosophila melanogaster. This review summarizes the progress in fly lipid droplet research along four main avenues: the role of lipid droplets in fat storage homeostasis, the control of lipid droplet structure, the lipid droplet surface as a dynamic protein-association platform, and lipid droplets as mobile organelles. Moreover, the research potential of the fruit fly model is discussed with respect to the prevailing general questions in lipid droplet biology. PMID- 21620888 TI - The interaction between HCV and nuclear receptor-mediated pathways. AB - Hepatitis C virus (HCV) is presently the leading indication for liver transplantation in Western countries. Treatment for HCV infection includes a combination of pegylated interferon and ribavirin, which produces highly variable response rates. This reflects the lack of information regarding the roles of host and viral components during viral pathogenesis. Vital processes regulated by the liver, including metabolism, lipid homeostasis, cellular proliferation, and the immune response, are known to be systematically dysregulated as a result of persistent HCV infection. Nuclear receptors and their ligands are recognized as indispensable regulators of liver homeostasis. Pathways mediated by the nuclear receptor superfamily have been shown to be profoundly disrupted during HCV infection, leading to an increased importance in elucidating the exact nature of this complex relationship. Expanded understanding of the role of nuclear receptors in HCV infection may therefore be an essential step in the search for a more universally effective treatment. PMID- 21620890 TI - A novel algorithm to generate kymographs from dynamic axons for the quantitative analysis of axonal transport. AB - The biological and clinical relevance of axonal transport has driven the development of a variety of new approaches to its study, including the generation of fluorescence or brightfield movies of moving cargoes within axons. Kymograph analysis is a simple and effective tool used to analyze axonal transport in neurons. Typically, kymographs are built by having a user trace the path of the axon in one frame of a time-lapse movie and extracting intensity profiles from subsequent frames along that path. This method cannot accommodate movies in which translation of the axon, or changes in axonal orientation or geometry, occur. Both are frequently observed in long-term movies of neurons, both in vitro and in vivo. To solve this problem and automate the creation of kymographs from these movies, we developed a two step algorithm. The first step implemented a simple image registration algorithm that aligned axons based on identification of a reference point on the axon in each image. The second step used a Hough transformation (HT) to automatically detect the axonal contour in each frame. Intensity profiles along this contour were then used to construct a kymograph. This algorithm was able to build an accurate kymograph of mitochondrial and actin transport in dynamic cultured sensory neurons, which were not amenable to previously used analytical methods. Although developed as a tool for analyzing transport, this algorithm is easily modified to analyze movies for the directionality and speed of axonal outgrowth, another metric of interest to neuroscientists. PMID- 21620891 TI - An automated calibration method for non-see-through head mounted displays. AB - Accurate calibration of a head mounted display (HMD) is essential both for research on the visual system and for realistic interaction with virtual objects. Yet, existing calibration methods are time consuming and depend on human judgements, making them error prone, and are often limited to optical see-through HMDs. Building on our existing approach to HMD calibration Gilson et al. (2008), we show here how it is possible to calibrate a non-see-through HMD. A camera is placed inside a HMD displaying an image of a regular grid, which is captured by the camera. The HMD is then removed and the camera, which remains fixed in position, is used to capture images of a tracked calibration object in multiple positions. The centroids of the markers on the calibration object are recovered and their locations re-expressed in relation to the HMD grid. This allows established camera calibration techniques to be used to recover estimates of the HMD display's intrinsic parameters (width, height, focal length) and extrinsic parameters (optic centre and orientation of the principal ray). We calibrated a HMD in this manner and report the magnitude of the errors between real image features and reprojected features. Our calibration method produces low reprojection errors without the need for error-prone human judgements. PMID- 21620892 TI - Primary culture of cellular subtypes from postnatal mouse for in vitro studies of oxygen glucose deprivation. AB - One of the most widely utilized in vitro models of ischemia or oxygen glucose deprivation (OGD) is the hippocampal organotypical culture (HOTC). The HOTC is used not only for the study of the mechanisms of cell death, but also has been the cornerstone of synaptic physiology. Although the intact nature of the HOTC is one of its primary advantages, some studies require a dissociated preparation in order to distinguish cell type specific responses. Typically, primary dissociated neuronal cultures are prepared from embryonic tissue. Since the HOTC is prepared from postnatal pups, we wanted to establish a primary culture of hippocampus from postnatal pups to parallel our studies in the HOTC preparation. Mixed cultures were prepared by enzymatic dissociation of hippocampus from 7-day-old mouse pups. These cultures responded to OGD with a time course of delayed cell death that was similar to that reported in HOTC. Dual label immunocytochemical staining revealed that neurons, but not astrocytes, were dying from apoptosis following OGD. To examine this vulnerability further, we also prepared neuronal enriched cultures by treating mixed cultures with cytosine-beta-d-arabinofuranoside (CBA). These neuronal cultures appear to be even more sensitive to OGD. In addition, we have established primary astrocyte-enriched cultures from the same age pups to examine the vulnerability of astrocytes to OGD. These three culture preparations are useful for comparison of the responses of the two major cell types in the same culture, and the enriched cultures will allow biochemical, electrophysiological and molecular studies of homogenous cell populations. PMID- 21620893 TI - IL-15 promotes osteoclastogenesis via the PLD pathway in rheumatoid arthritis. AB - Osteoclastogenesis plays an important role in joint destruction in rheumatoid arthritis (RA). IL-15 is a pleiotropic proinflammatory cytokine that appears to help mediate the pathological bone loss. This study was undertaken to explore the signaling molecules essential for osteoclastogenesis mediated by IL-15 in rheumatoid synovial fibroblasts. Expression of phospholipase D1 (PLD1) and osteoclast-related gene expression in synovial tissues and their modulation by treatment with IL-15 and different inhibitors in synovial fibroblasts of RA patients were evaluated using immunohistochemistry and quantitative polymerase chain reaction. The levels of IL-15 in serum and synovial fluid were measured by ELISA. The effects of IL-15 and phosphatidic acid (PA) on osteoclast formation were evaluated in cocultures of rheumatoid synovial fibroblasts and peripheral blood monocytes or monocytes alone in the presence of M-CSF and RANKL. The levels of RANKL and PLD1 but not PLD2 were upregulated significantly by IL-15, and the RANKL level was significantly upregulated by PA in rheumatoid synovial fibroblasts. Blocking PA production with 1-butanol and siRNA against PLD1 significantly inhibited the IL-15-stimulated expression of RANKL and PLD1. IL-15 levels were significantly higher in serum and synovial fluid from patients with RA than in osteoarthritis patients and healthy controls. IL-15 and PA induced osteoclast formation through the mitogen-activated protein kinases (MAPKs) and NF kappaB signaling pathways. Activation of PLD1 contributes to IL-15-mediated osteoclastogenesis via the MAPKs and NF-kappaB signaling pathways in rheumatoid synovial fibroblasts. Our data suggest that PLD1 might be an efficient therapeutic strategy for preventing bone destruction in rheumatoid arthritis. PMID- 21620894 TI - Low CTLA-4 expression in CD4+ helper T-cells in patients with fulminant type 1 diabetes. AB - Fulminant type 1 diabetes is a novel subtype of type 1 diabetes characterized by a remarkably abrupt onset of insulin-deficient hyperglycemia. An accelerated immune reaction has been suggested as the cause of markedly rapid beta cell loss in this disease, but the precise mechanism has not been clarified. We analyzed the expression of cytotoxic T lymphocyte antigen 4 (CTLA-4) in CD4(+) helper T cells in 16 patients with fulminant type 1 diabetes, 14 patients with type 1A diabetes, 10 patients with type 2 diabetes and 20 normal control subjects. There was a significant reduction in CTLA-4 expression in CD4(+) helper T-cells from patients with fulminant type 1 diabetes (P<0.05) compared with the other three groups. Low CTLA-4 expression was also observed in both CD4(+)CD25(high) T-cells and CD4(+)CD25(-) T-cells. There was a significant negative correlation between the proliferation of CD4(+)CD25(-) T-cells and the levels of CTLA-4. Intracellular expression of CTLA-4 in CD4(+) helper T-cells was not correlated with two CTLA-4 polymorphisms. In conclusion, the expression of CTLA-4 in CD4(+) helper T-cells was low in patients with fulminant type 1 diabetes. PMID- 21620895 TI - Isolation and characterization of a human antibody fragment specific for Ts1 toxin from Tityus serrulatus scorpion. AB - Scorpion stings are a common event that occurs in tropical and subtropical areas of the world, being a public health problem in certain countries. In most places, medical treatment relays on antivenoms obtained from the sera of hyper-immunized horses, however some efforts are being made to prepare specific antibodies of human origin, using phage display methodology. This communication describes the strategy followed for obtaining a protective human single chain antibody (scFv) capable of partially neutralizing the effect of Ts1, the major toxin isolated from the venom of the Brazilian scorpion Tityus serrulatus. Phage display technique allowed the isolation of scFv 15e from a human library of antibodies, after four rounds of selection against Ts1. This clone codes for 124 amino acids belonging to the family VH6 and 114 amino acids of family VK4. This scFv also recognizes toxins from the scorpions Tityus packyurus and Tityus cambridgei from the Amazonian region. Mice challenged with a LD(50) of Ts1 in the presence of this scFv were substantially resistant to intoxication. ScFv 15e is a leading compound for the development of better anti-scorpion antidotes. PMID- 21620896 TI - An ELISA for detection of trout antibodies to viral haemorrhagic septicemia virus using recombinant fragments of their viral G protein. AB - An enzyme linked immunosorbent assay (ELISA) method to study serum antibodies to viral haemorrhagic septicemia virus (VHSV) was designed by using recombinant fragments of their G protein. By using this fragment-ELISA, we describe the binding of antibodies against recombinant G fragments of 45-445 amino acids present in VHSV-hyperimmunized trout sera. Fragments were designed by taking into account their tridimensional pH-dependent structure and functional domains. Sera were obtained from hyperimmunized trout following 4-5 intraperitoneal injections of VHSV antigens by using Freund's or saponin adjuvants. Sera from different hyperimmunized trout differed quantitatively rather than qualitatively in their recognition of solid-phase frg11 (56-110), frg12 (65-109), frg13 (97-167), frg14 (141-214), frg15 (65-250), frg16 (252-450) and G (G21-465) by Western blot and ELISA. However, titres were higher when using frg11, frg15 or frg16, rather than G21-465, suggesting higher accessibility to G epitopes. Further knowledge of the antigenicity of the G protein of rhabdoviruses by using fragments might be used to improve current vaccines. On the other hand, they might be used to dissect the trout antibody response to VHSV infections, to complement in vitro neutralizing assays, and/or to quantitate anti-VHSV antibodies in VHSV-infected/vaccinated trout, other fish and/or other body fluids such as mucus. PMID- 21620897 TI - Detection of nine respiratory RNA viruses using three multiplex RT-PCR assays incorporating a novel RNA internal control transcript. AB - Real-time PCR is a significant improvement over viral isolation and immunofluorescence for routinely detecting respiratory viruses. We developed three real-time internally controlled multiplex RT-PCR assays for detecting nine respiratory viruses. An internal control transcript consisting of a chimeric plasmid was synthesised and incorporated into each multiplex to monitor amplification efficiency, including inhibition. Each multiplex assay was developed on the Rotor-Gene 3000 and evaluated using RNA extracts from 126 nasopharyngeal aspirates from 112 pre-term infants. All 44/126 (35%) samples positive by immunofluorescence were confirmed by multiplex RT-PCR. Additionally, respiratory syncytial virus RNA was detected in 5 samples, influenza A virus RNA in 2 samples and thirteen (10%) dual infections by multiplex RT-PCR were noted. Inclusion of the RNA internal control did not affect the amplification efficiency of the target sequences and only 2 of 1256 (0.2%) samples tested over a 12 month period were inhibitory. Together with the improved sensitivity of the internally controlled multiplex RT-PCR assays over the older technology and the ability to detect co-infections, the internal control monitored the efficiency of both the RT and PCR steps and indicated inhibition, saving time and costs on running duplicate samples with a "spiked" inhibition control. PMID- 21620898 TI - Multiplex RT-PCR detection and microarray typing of vesicular disease viruses. AB - A vesicular disease multiplex reverse transcription (RT)-PCR with an accompanying microarray assay was developed for simultaneous detection and typing of foot-and mouth disease virus (FMDV) and vesicular stomatitis virus (VSV), and for the detection of swine vesicular disease virus (SVDV) and vesicular exanthema of swine virus (VESV). The multiplex RT-PCR successfully detected viral RNA from a collection of 49 strains of vesicular viruses, including multiple strains from all seven serotypes of FMDV and both serotypes of VSV. The multiplex RT-PCR was also able to produce amplified products from the RNA genome of all four viruses simultaneously in mixed samples. An indirect (post-PCR labelling) amplicon labelling method and a direct (concurrent labelling with PCR) amplicon labelling method were compared for the purpose of microarray detection and typing. Accurate detection and typing was achieved with all strains tested in the microarray assay which utilized 163 virus- and serotype-specific probes. It was observed that microarray increased detection for some samples compared to using multiplex RT PCR alone. This was most likely due to signal amplification resulting from fluorescent labelling. The limit of detection of the microarray assay was as low as 4.6TCID(50)/mL for FMDV. No amplification products or microarray reactivity was observed with non-target livestock pathogens tested or with samples collected from healthy cattle, sheep and pigs. All FMDV and VSV serotypes were detected as early as 2 days post-inoculation from oral swabs obtained from cattle infected experimentally. PMID- 21620899 TI - Development of hepatitis C virus chimeric replicons for identifying broad spectrum NS3 protease inhibitors. AB - Several potent inhibitors of hepatitis C virus (HCV) NS3/4A protease have been identified that show great clinical potential against genotype 1. Due to the tremendous genetic diversity that exists among HCV isolates, development of broad spectrum inhibitors is challenging. With a limited number of lab strains available for preclinical testing, new tools are required for assessing protease inhibitor activity. We developed a chimeric replicon system for evaluating NS3 protease inhibitor activity against naturally occurring isolates. NS3/4A genes were cloned from the plasma of HCV-infected individuals and inserted into lab strain replicons, replacing the native sequences. The chimeric reporter replicons were transfected into Huh 7.5 cells, their replication monitored by luciferase assays, and their susceptibilities to inhibitors determined. Viable chimeras expressing heterologous genotypes 1, 2, 3, and 4 protease domains were identified that exhibited varying susceptibilities to inhibitors. Protease inhibitor spectrums observed against the chimeric replicon panel strongly correlated with published enzymatic and clinical results. This cell-based chimeric replicon system can be used to characterize the activities of protease inhibitors against diverse natural isolates and may improve the ability to predict dose and clinical efficacy. PMID- 21620900 TI - Sex-specific neuroendocrine and behavioral phenotypes in hypomorphic Type II Neuregulin 1 rats. AB - Neuregulin 1 (NRG1) is an important growth factor involved in the development and plasticity of the central nervous system. Since its identification as a susceptibility gene for schizophrenia, several transgenic mouse models have been employed to elucidate the role NRG1 may play in the pathogenesis of psychiatric disease. Unfortunately very few studies have included females, despite the fact that some work suggests that the consequences of disrupted NRG1 expression may be sex-specific. Here, we used Nrg1 hypomorphic (Nrg1(Tn)) Fischer rats to demonstrate sex-specific changes in neuroendocrine and behavioral phenotypes as a consequence of reduced Type II NRG1 expression. We have previously shown that male Nrg1(Tn) rats have increased basal corticosterone levels, and fail to habituate to an open field despite normal overall levels of locomotor activity. The current studies show that, in contrast, female Nrg1(Tn) rats exhibit enhanced suppression of corticosterone levels following an acute stress, reduced locomotor activity, and enhanced habituation to novel environments. Furthermore, we also show that female, but not male, Nrg1(Tn) rats have impaired prepulse inhibition. Finally, we provide evidence that sex-specific changes are not likely attributable to major disruptions in the hypothalamic-pituitary-gonadal axis, as measures of pubertal onset, estrous cyclicity, and reproductive capacity were unaltered in female Nrg1(Tn) rats. Our results provide further support for both the involvement of NRG1 in the control of hypothalamic-pituitary-adrenal axis function and the sex-specific nature of this relationship. PMID- 21620901 TI - ICV STZ induced impairment in memory and neuronal mitochondrial function: A protective role of nicotinic receptor. AB - The present study was planned to evaluate the cholinergic influence on mitochondrial activity and neurodegeneration associated with impaired memory in intracerebroventricular (ICV) streptozotocin (STZ) treated rats. STZ (3mg/kg), administered ICV twice with an interval of 48h between the two doses, showed significant impairment in spatial memory tested by water maze test 14 days after first dose without altering blood glucose level and locomotor activity. Animals were sacrificed on 21st day of ICV administration. STZ significantly increased malondialdehyde (MDA), reactive oxygen species (ROS), Ca(2+) ion influx, caspase 3 activity and decreased glutathione (GSH) level. Acetylcholinesterase inhibitors tacrine and donepezil (5mg/kg, PO) pretreatment significantly prevented STZ induced memory deficit, oxidative stress, Ca(2+) influx and caspase-3 activity. Carbachol, a muscarinic cholinergic agonist (0.01mg/kg, SC) did not show any significant effect on ROS generation, Ca(2+) ion influx and caspase-3 activity. While nicotinic cholinergic agonist, nicotine, significantly attenuated ICV STZ induced mitochondrial dysfunction and caspase-3 activity. The results indicate that instead of muscarinic receptors nicotinic receptors may be involved in neuroprotection by maintaining mitochondrial functions. PMID- 21620902 TI - Transcript-level responses of Plasmodium falciparum to thiostrepton. AB - The antimalarial activity of the antibiotic thiostrepton has long been attributed to inhibition of apicoplast protein synthesis through binding of apicoplast ribosomal RNA. However, the kinetics of parasite death upon thiostrepton treatment differ from those seen for other inhibitors of apicoplast housekeeping functions. We have analysed global changes in gene expression of the malaria parasite, Plasmodium falciparum, in an attempt to shed light on the responses of the parasite to this drug. Our results indicate a delay in gene expression profiles of thiostrepton-treated parasites. A small number of genes appear to be regulated outside of this trend; our data suggest a response from genes encoding components of the mitochondrial translational machinery, while little response is seen from genes encoding apicoplast-targeted proteins. Our findings are consistent with an effect of thiostrepton on mitochondrial protein synthesis, and thus warrant a re-evaluation of the target of thiostrepton in Plasmodium. They also provide some suggestion of mitochondrion-nucleus signalling in the parasite. PMID- 21620903 TI - Abnormally decreased NO and augmented CO production in islets of the leptin deficient ob/ob mouse might contribute to explain hyperinsulinemia and islet survival in leptin-resistant type 2 obese diabetes. AB - The role of the gaseous messengers NO and CO for beta-cell function and survival is controversial. We examined this issue in the hyperglycemic-hyperinsulinemic ob/ob mouse, an animal model of type 2 obese diabetes, by studying islets from obese vs lean mice regarding glucose-stimulated insulin release in relation to islet NO and CO production and the influence of modulating peptide hormones. Glucose-stimulated increase in ncNOS-activity in incubated lean islets was converted to a decrease in ob/ob islets associated with markedly increased insulin release. Both types of islets displayed iNOS activity appearing after ~60 min in high-glucose. In ob/ob islets the insulinotropic peptides glucagon, GLP-1 and GIP suppressed NOS activities and amplified glucose-stimulated insulin release. The insulinostatic peptide leptin induced the opposite effects. Suppression of islet CO production inhibited, while stimulation amplified glucose stimulated insulin release. Nonincubated isolated islets from young and adult obese mice displayed very low ncNOS and negligible iNOS activity. In contrast, production of CO, a NOS inhibitor, was impressively raised. Glucose injections induced strong activities of islet NOS isoforms in lean but not in obese mice and confocal microscopy revealed iNOS expression only in lean islets. Islets from ob/ob mice existing in a hyperglycemic in vivo milieu maintain elevated insulin secretion and protection from glucotoxicity through a general suppression of islet NOS activities achieved by leptin deficiency, high CO production and insulinotropic cyclic-AMP-generating hormones. Such a beneficial effect on islet function and survival might have its clinical counterpart in human leptin resistant type 2 obese diabetes with hyperinsulinemia. PMID- 21620904 TI - Rapid hybrid de novo assembly of a microbial genome using only short reads: Corynebacterium pseudotuberculosis I19 as a case study. AB - Due to the advent of the so-called Next-Generation Sequencing (NGS) technologies the amount of monetary and temporal resources for whole-genome sequencing has been reduced by several orders of magnitude. Sequence reads can be assembled either by anchoring them directly onto an available reference genome (classical reference assembly), or can be concatenated by overlap (de novo assembly). The latter strategy is preferable because it tends to maintain the architecture of the genome sequence the however, depending on the NGS platform used, the shortness of read lengths cause tremendous problems the in the subsequent genome assembly phase, impeding closing of the entire genome sequence. To address the problem, we developed a multi-pronged hybrid de novo strategy combining De Bruijn graph and Overlap-Layout-Consensus methods, which was used to assemble from short reads the entire genome of Corynebacterium pseudotuberculosis strain I19, a bacterium with immense importance in veterinary medicine that causes Caseous Lymphadenitis in ruminants, principally ovines and caprines. Briefly, contigs were assembled de novo from the short reads and were only oriented using a reference genome by anchoring. Remaining gaps were closed using iterative anchoring of short reads by craning to gap flanks. Finally, we compare the genome sequence assembled using our hybrid strategy to a classical reference assembly using the same data as input and show that with the availability of a reference genome, it pays off to use the hybrid de novo strategy, rather than a classical reference assembly, because more genome sequences are preserved using the former. PMID- 21620905 TI - Reference genes for quantitative, reverse-transcription PCR in Bacillus cereus group strains throughout the bacterial life cycle. AB - Quantitative reverse-transcription PCR (RT-qPCR) has become a major tool to better understand the biology and pathogenesis of bacteria. One prerequisite of valid RT-qPCR data is their proper normalization to stably expressed reference genes. To identify and evaluate reference genes suitable for normalization of gene expression data in Bacillus cereus group strains, mRNA levels of eleven candidate reference genes (rpsU, nifU, udp (UDP-N-acetylglucosamine 2-epimerase), BT9727_5154/BC_5475, BT9727_4034/BC_4293, BT9727_4549/BC_4813, pspA, gatB_Yqey (gatB_Yqey domain containing protein), helicase (SWF/SNF family protein), adk and pta) and a target gene (BT9727_3305/BC3547+BC3546) were quantified by RT-qPCR at different time points throughout the entire life cycle of the wild-type B. cereus ATCC 14579 and Bacillus thuringiensis subsp. konkukian 97-27, a phylogenetically closely related strain to Bacillus anthracis. The programs geNorm and Normfinder were used to calculate expression stabilities and identified the genes gatB_Yqey, rpsU and udp as the most stably expressed reference genes. Compared to this combination or the sets of reference genes as recommended by geNorm or Normfinder, normalization using a traditional housekeeping gene (adk) alone resulted in significantly different gene expression results and in a significant overestimation of the target gene transcription. Normalization of the data to the reference gene gatB_Yqey alone showed no or only small differences to the reference gene combinations indicating that gatB_Yqey may be used as a single reference gene when investigating rather large changes in mRNA transcription. Otherwise, a combination of the stably expressed reference genes is recommended. In conclusion, the present study underlines the importance of normalization to stably expressed reference genes and presents valid endogenous controls suitable for normalization of transcriptional data throughout the life cycle of B. cereus group strains. PMID- 21620906 TI - Interplay of neuronal processes during response inhibition: results from a combined event-related potentials (ERPs)/transcranial magnetic stimulation (TMS) study on methylphenidate. AB - The neuronal processes underlying response inhibition are often studied using either event-related potentials (ERPs) or by applying transcranial magnetic stimulation (TMS) to investigate excitatory and inhibitory processes in the motor system. We performed a more refined analysis of response inhibition by combining both approaches with the aim of identifying an interplay between ERPs and TMS parameters. During a go/nogo task, motor system excitability was measured using TMS single and double pulses and brain electrical activity was recorded in healthy adults (n=14). Each participant completed two testing sessions, once on placebo and once on methylphenidate (double-blind, crossover design). Studying the effects of methylphenidate served as an example application for this combined approach. Developing regression models, inhibition-related TMS measures (e.g., short intracortical inhibition) and the contingent negative variation explained about 85% of the variance of the nogo-P3 under both MPH and placebo medication. The smaller the inhibitory effect in the motor system, the more terminal response control was required and the more resources were allocated for the evaluation of the inhibitory process, respectively, as indicated by a larger P3. Thus, an interplay between processes in the motor system (cortex) and control processes with sources in the prefrontal cortex and the anterior cingulate cortex (ACC) may take place, acting complementarily to facilitate a correct nogo-response. While ERPs rather represent initiation and monitoring of inhibitory processes and response control, motor inhibition may be best analyzed using TMS. A combined ERP/TMS analysis may allow for the development of distinct models concerning the interplay of processes involved in response inhibition. PMID- 21620907 TI - How negative affect influences neural control processes underlying the resolution of cognitive interference: an event-related fMRI study. AB - In this event-related fMRI study, we sought to investigate the influence of negative affect on the processing of two kinds of cognitive interference: Stroop interference and oddball interference. For our purpose, we adopted an oddball variant of the Stroop task in which Stroop-interference and oddball interference conditions were created by presenting incongruent and rarely occurring word meanings, respectively. Immediately preceding the target stimuli, we presented pictures of the International Affective Picture System which were either emotionally negative and arousing or emotionally neutral, providing two affective conditions under which the cognitive task was administered. Both the behavioral and the neuroimaging data exhibited an interaction effect between emotional and cognitive condition. First, the emotion induction selectively impaired behavioral performance on interference trials while behavioral measures on non-interference trials were roughly identical in both emotional conditions. Second, in the negative emotional condition there was incremental interference-related activation in control-related regions (fronto-parietal cortices). Taken together, findings suggest that negative affect specifically disturbs the neural control processes that in a neutral affective state allow to select task-relevant information and to shield its processing from task-irrelevant distraction. Accordingly, agents in a negative affective state have to exert enhanced control efforts to resolve cognitive interference. Additional connectivity analyses revealed that a negative coupling between lateral PFC on the one hand and amygdala and OFC on the other is related to enhanced interference resolution which can be tentatively interpreted as evidence that emotional regulation is an integrated part of an agent's efforts to preserve cognitive performance in affective situations. PMID- 21620908 TI - Toward the development of a hepatitis E vaccine. AB - Hepatitis E virus (HEV) causes large epidemics of enterically transmitted acute hepatitis and accounts for a majority of sporadic acute hepatitis in endemic countries. Due to a very high mortality rate among infected pregnant women and substantial morbidity, disability and costs associated with hepatitis E, concerted efforts are being made to develop an efficacious vaccine. Experimental vaccines, based on recombinant proteins derived from the capsid gene of HEV, have been shown efficacious in pre-clinical trials in macaques conferring cross protection against various genotypes. Two vaccine candidates, the rHEV vaccine expressed in baculovirus and the HEV 239 vaccine, expressed in Escherichia coli, were successfully evaluated in Phase II/III trials. However, at this time no approved vaccine against hepatitis E is commercially available. PMID- 21620909 TI - The expanding field of plant virus ecology: historical foundations, knowledge gaps, and research directions. AB - Plant viruses are widespread in nature, where they operate in intimate association with their hosts and often with vectors. Most research on plant viruses to the present has focused on agricultural systems (agronomic and horticultural) and viruses that are pathogenic. Consequently, there is a dearth of fundamental information about plant virus dynamics in natural ecosystems and how they might differ from or be influenced by virus interactions in managed systems. Key questions include under what conditions the influence of virus on host fitness is negative, neutral, or positive and the extent to which this relationship is influenced by ecosystem properties. To address these critical knowledge gaps, the expanding field of plant virus ecology seeks to examine (i) the ecological roles of plant-associated viruses and their vectors in managed and unmanaged ecosystems and (ii) the reciprocal influence of ecosystem properties on the distribution and evolution of plant viruses and their vectors. In this work, plant virus ecology draws on the achievements of epidemiology and extends the research focus to new ecological arenas. Here we provide an historical perspective and highlight key issues and emerging research directions. We suggest that there is broad need to (i) integrate consideration of plant viruses into ecological research and theory, in which viruses have generally been overlooked, and (ii) to expand ecological perspectives in virology to include new methods and disciplines in ecology, such as ecosystem ecology. Studies of plant-virus-vector interactions in nature offer both opportunities and challenges that will ultimately produce multi-faceted understanding of the role of viruses in shaping ecological and evolutionary dynamics. PMID- 21620911 TI - Human serum albumin-thioredoxin fusion protein with long blood retention property is effective in suppressing lung injury. AB - Thioredoxin (Trx) is a redox-active protein with anti-inflammatory effects but with a short half life of 1 h. Genetic fusion of Trx to human serum albumin (HSA) extended its half life without causing significant loss of its biological activities. HSA-Trx caused a decrease in the number of cells in brochoalveolar lavage fluid, the wet/dry ratio and the inflammation at the respiratory tract of the ovalbumin (OVA) induced lung injury model mouse. Three intraperitoneal doses of Trx alone produced the same extent of suppression of those three detrimental effects of OVA as one intravenous dose of HSA-Trx. Inhibition experiments confirmed that reactive oxygen species (ROS) and reactive nitrogen species (RNS) involved in the progression of the injury. HSA-Trx inhibited the production of ROS as confirmed in the EPR experiment, but lung tissue staining suggested that induced nitrogen oxide synthase (iNOS) was not suppressed by the fusion protein. Instead, the production of nitrotyrosine, 8-nitro-cGMP, and 8-hydroxy-2' deoxyguanosine downstream to the iNOS has been inhibited. This suggested that HSA Trx produced lung protection effect via different mechanisms from Trx alone. HSA Trx retains the biological properties of Trx thus has great potential in treating oxidative stress related diseases. PMID- 21620910 TI - Polymer delivery systems for site-specific genome editing. AB - Triplex-forming peptide nucleic acids (PNAs) can be used to coordinate the recombination of short 50-60bp "donor DNA" fragments into genomic DNA, resulting in site-specific correction of genetic mutations or the introduction of advantageous genetic modifications. Site-specific gene editing in hematopoietic stem and progenitor cells (HSPCs) could result in the treatment or cure of inherited disorders of the blood such as beta-thalassemia or sickle cell anemia. Gene editing in HSPCs and differentiated T cells could also help combat HIV infection by modifying the HIV co-receptor CCR5, which is necessary for R5-tropic HIV entry. However, translation of genome modification technologies to clinical practice is limited by challenges in intracellular delivery, especially in difficult-to-transfect hematolymphoid cells. Here, we review the use of engineered biodegradable polymer nanoparticles for site-specific genome editing in human hematopoietic cells, which represent a promising approach for ex vivo and in vivo gene therapy. PMID- 21620912 TI - Drug-loaded polyelectrolyte microcapsules for sustained targeting of cancer cells. AB - In this review we will overview novel nanotechnological nanocarrier systems for cancer therapy focusing on recent development in polyelectrolyte capsules for targeted delivery of antineoplastic drugs against cancer cells. Biodegradable polyelectrolyte microcapsules (PMCs) are supramolecular assemblies of particular interest for therapeutic purposes, as they can be enzymatically degraded into viable cells, under physiological conditions. Incorporation of small bioactive molecules into nano-to-microscale delivery systems may increase drug's bioavailability and therapeutic efficacy at single cell level giving desirable targeted therapy. Layer-by-layer (LbL) self-assembled PMCs are efficient microcarriers that maximize drug's exposure enhancing antitumor activity of neoplastic drug in cancer cells. They can be envisaged as novel multifunctional carriers for resistant or relapsed patients or for reducing dose escalation in clinical settings. PMID- 21620913 TI - Compensatory beliefs and intentions contribute to the prediction of caloric intake in dieters. AB - One cognitive process that impacts dieters' decision to indulge is the activation of compensatory beliefs. Compensatory beliefs (CBs) are convictions that the consequences of engaging in an indulgent behaviour (eating cake) can be neutralized by the effects of another behaviour (skipping dinner). Using experience sampling methodology, this study hypothesized that, in addition to the cognitive processes associated with restraint and disinhibition, compensatory thinking contributes to the prediction of caloric intake. Results indicated that higher scores on CB, CI and TFEQ-D predicted a greater number of portions eaten signifying that, along with disinhibition, compensatory thinking predicts caloric intake in dieters. PMID- 21620914 TI - Virucidal activity of a scorpion venom peptide variant mucroporin-M1 against measles, SARS-CoV and influenza H5N1 viruses. AB - Outbreaks of SARS-CoV, influenza A (H5N1, H1N1) and measles viruses in recent years have raised serious concerns about the measures available to control emerging and re-emerging infectious viral diseases. Effective antiviral agents are lacking that specifically target RNA viruses such as measles, SARS-CoV and influenza H5N1 viruses, and available vaccinations have demonstrated variable efficacy. Therefore, the development of novel antiviral agents is needed to close the vaccination gap and silence outbreaks. We previously identified mucroporin, a cationic host defense peptide from scorpion venom, which can effectively inhibit standard bacteria. The optimized mucroporin-M1 can inhibit gram-positive bacteria at low concentrations and antibiotic-resistant pathogens. In this investigation, we further tested mucroporin and the optimized mucroporin-M1 for their antiviral activity. Surprisingly, we found that the antiviral activities of mucroporin-M1 against measles, SARS-CoV and influenza H5N1 viruses were notably increased with an EC50 of 7.15 MUg/ml (3.52 MUM) and a CC50 of 70.46 MUg/ml (34.70 MUM) against measles virus, an EC50 of 14.46 MUg/ml (7.12 MUM) against SARS-CoV and an EC50 of 2.10 MUg/ml (1.03 MUM) against H5N1, while the original peptide mucroporin showed no antiviral activity against any of these three viruses. The inhibition model could be via a direct interaction with the virus envelope, thereby decreasing the infectivity of virus. This report provides evidence that host defense peptides from scorpion venom can be modified for antiviral activity by rational design and represents a practical approach for developing broad-spectrum antiviral agents, especially against RNA viruses. PMID- 21620915 TI - Influenza vaccine coverage, influenza-associated morbidity and all-cause mortality in Catalonia (Spain). AB - The objective of this work was to study the behaviour of influenza with respect to morbidity and all-cause mortality in Catalonia, and their association with influenza vaccination coverage. The study was carried out over 13 influenza seasons, from epidemiological week 40 of 1994 to week 20 of 2007, and included confirmed cases of influenza and all-cause mortality. Two generalized linear models were fitted: influenza-associated morbidity was modelled by Poisson regression and all-cause mortality by negative binomial regression. The seasonal component was modelled with the periodic function formed by the sum of the sinus and cosines. Expected influenza mortality during periods of influenza virus circulation was estimated by Poisson regression and its confidence intervals using the Bootstrap approach. Vaccination coverage was associated with a reduction in influenza-associated morbidity (p<0.001), but not with a reduction in all-cause mortality (p=0.149). In the case of influenza-associated morbidity, an increase of 5% in vaccination coverage represented a reduction of 3% in the incidence rate of influenza. There was a positive association between influenza associated morbidity and all-cause mortality. Excess mortality attributable to influenza epidemics was estimated as 34.4 (95% CI: 28.4-40.8) weekly deaths. In conclusion, all-cause mortality is a good indicator of influenza surveillance and vaccination coverage is associated with a reduction in influenza-associated morbidity but not with all-cause mortality. PMID- 21620916 TI - High burden of tick-borne encephalitis in Slovenia--challenge for vaccination policy. AB - Slovenia is one of the countries with the highest reported incidence rates of tick-borne encephalitis (TBE). Vaccination uptake is low, estimated to be 12.4%. TBE surveillance data for the last 20 years were analysed. Though nearly all of Slovenia is endemic for TBE with national yearly incidence rates up to 26.7/100,000, we showed that two regions (Gorenjska and Koroska) were much more affected than other seven regions, with annual incidence rates up to 57.2/100,000 and 76.9/100,000 population, respectively. In the last decade, there was a shift in the age distribution of reported TBE cases to the older age groups, which resulted in the highest age-specific incidence rates nationally in 55-64 age group (up to 33.4/100,000 in 2006). To reduce this high burden of TBE, ideally the whole population of Slovenia should be offered free of charge vaccination against TBE. Alternatively, in view of limited resources available, sensible approach would be increasing vaccination coverage of the general population using social marketing and increasing TBE awareness, and in addition, offering free of charge vaccination to the most affected groups. The following priority target groups should be considered to be prospectively covered with free of charge vaccination: (1) 45-69 years old individuals in the two most affected regions (Gorenjska, Koroska), (2) the remaining age groups in the two most affected regions, (3) 45-69 years old individuals in the region with the next highest TBE incidence rates (Ljubljana), and (4) individuals 45-69 years old in all remaining Slovenian regions. PMID- 21620917 TI - Characterization of the RepliVax platform for replication-defective flavivirus vaccines. AB - RepliVax, a novel replication-defective vaccine platform has recently been described as a suitable means of generating potent vaccines targeting flaviviruses. In this study, we directly compared attenuation, immunogenicity and efficacy of several prototype RepliVax constructs to available, well characterized live attenuated (LAV) and inactivated (INV) flavivirus vaccine controls in mice and hamsters. Other important aspects of general mechanisms and properties of RepliVax vaccines were also studied. The prototypes were found to be nonpathogenic in sensitive suckling mouse neurovirulence tests, and highly immunogenic and efficacious in mice and hamsters, with evidence that immunogenicity can be comparable to LAV controls in terms of both magnitude and durability of response. Our data also suggest that choice of inoculation route can be beneficial for maximizing RepliVax immunogenicity. Additionally, different vaccine constructs can be administered as cocktail formulations without compromising immunogenicity of individual components. RepliVax constructs were determined to induce a Th1 biased immune response, similar to LAVs, and different from INV inducing a Th2 type response. The results presented validate the utility of the RepliVax platform for development of novel flavivirus vaccines. PMID- 21620918 TI - Assessment of combinations of antiandrogenic compounds vinclozolin and flutamide in a yeast based reporter assay. AB - Humans are exposed to a combination of various substances such as cosmetic ingredients, drugs, biocides, pesticides and natural-occurring substances in food. The combined toxicological effects of two or more substances can simply be additive on the basis of response-addition, or it can be greater (synergistic) or smaller (antagonistic) than this. The need to assess combined effects of compounds with endocrine activity is currently discussed for regulatory risk assessment. We have used a well described yeast based androgen receptor transactivation assay YAS to assess the combinatorial effects of vinclozolin and flutamide; both mediating antiandrogenicity via the androgen receptor. Both vinclozolin and flutamide were antiandrogens of similar potency in the YAS assay. In the concentration range tested the two antiandrogens vinclozolin and flutamide did not act synergistically. Concentration additivity was observed in the linear, non-receptor-saturated concentration range. At high concentrations of one of the two substances tested the contribution of the second at lower concentration levels was less than additive. The combined response of both compounds at high concentration levels was also less than additive (saturation effect). At concentration levels which did not elicit a response of the individual compounds, the combination of these compounds also did not elicit a response. PMID- 21620919 TI - A No Observable Adverse Effects Level (NOAEL) for pigs fed melamine and cyanuric acid. AB - Ingesting melamine adulterated milk products led to kidney stones in many infants in 2008. This differs from the renal failure caused by intratubular crystal formation after co-ingestion of melamine (MEL) and cyanuric acid (CYA) in adulterated pet foods in 2007. To better understand the potential risk of developing crystal nephropathy following co-ingestion of MEL and CYA, we fed 16 weanling pigs 0, 1, 3.3, 10, 33, or 100 mg/kg bw/day of each MEL and CYA, or 200 mg/kg bw/day of either compound individually for 7 days. Crystals were found in the renal medulla and cortex and urine sediments of all pigs fed both MEL and CYA each at 10 mg/kg bw/day (or greater). Crystals were also found in one of the two pigs fed 200 mg/kg bw/day MEL-only. In a 28 day study, 36 weanling pigs were fed 0, 1, or 3.3 mg/kg bw/day of MEL and CYA or 200 mg/kg bw/day MEL-only. Only one of the 3.3 mg/kg MEL and CYA pig kidneys contained crystals. The no-observed adverse-effect level (NOAEL) for pigs fed MEL and CYA for 28 days was concluded to be 1.0 mg/kg bw/day corresponding to 25 mg/kg (ppm) MEL and 25 mg/kg (ppm) CYA in dry feed. PMID- 21620920 TI - Acute and chronic effects of electroconvulsive treatment on oxidative parameters in schizophrenia patients. AB - Electroconvulsive therapy (ECT) is an effective treatment alternative for schizophrenia. Previous studies have already indicated the possible effects of oxidative stress in this disorder. However, there have been no previous studies evaluating the effects of ECT on the oxidative stress in these patients. We therefore aimed to investigate the acute and chronic effects of ECT on serum levels of oxidant and antioxidant molecules in schizophrenia patients (n=28). The serum MDA and CAT levels of the patients with schizophrenia were higher than that of the controls before ECT (n=20) but there was no significant difference in the serum NO and GSH levels of the patient groups compared to the controls. We found that the NO levels of the patients were higher than the controls in the group experiencing their first episode but not in the chronic group. There was a significant clinical improvement in the patients in terms of BPRS, SANS and SAPS reduction after the 9th ECT, but not the 1st ECT. Serum MDA levels were significantly reduced compared to the baseline after the 9th ECT session although there was no significant difference after the 1st session. Separate evaluation of the patient groups revealed that the significant MDA decrease following ECT was in the patients experiencing their first episode and not in the chronic group. No significant difference was noted in the serum levels of other oxidant and antioxidant molecules after either the 1st or 9th ECT session. These results suggest that ECT does not produce any negative effect on oxidative stress in patients with schizophrenia. PMID- 21620921 TI - Aggression and psychopharmacological treatments in major psychosis and personality disorders during hospitalisation. AB - BACKGROUND: A number of large-scale studies have shown that there is a relationship between many psychiatric disorders and aggression or violence. As no medication is currently approved for the treatment of aggression, pharmacotherapy (often involving drug combinations) is used on a trial-and-error basis with various degrees of response. METHOD: The study involved 244 in-patients aged 19 83 years (mean 41.9 +/- 11.3 SD). The Modified Overt Aggression Scale (MOAS) was used to assess any aggressive or violent behaviors occurring in the week before admission and upon discharge. Psychopathology was assessed using the Brief Psychiatric Rating Scales (BPRS). RESULTS: All of the patients showed a significant improvement (p<0.001) in mean weighted total MOAS scores at the end of the study, with no significant differences between the various drugs or combination therapies. The patients who received combination treatments including antidepressants showed a worsening in the weighted total MOAS score (18.46% +/- 114.31% SD); the patients who did not receive antidepressants had an improvement (13.61% +/- 257.36% SD) (p=0.0069). CONCLUSIONS: Multivariate testing of the variables age, gender, substance/alcohol abuse, the duration of hospitalisation, the administration of mood stabilisers, and the use of typical or atipical antipsychotics showed that the severity of the psychopathological picture correlated significantly with the presence of violence, whereas the effect of combined antidepressant treatment on violent behavior was only relative. PMID- 21620922 TI - The chemopreventive effect of dimethylthiourea against carmustine-induced myelotoxicity in rats. AB - The possible chemopreventive role of dimethylthiourea (DMTU) against carmustine (1,3-bis(2-chloroethyl)-1-nitrosourea, BCNU)-induced myelotoxicity was assessed through evaluation of apoptosis, lipid peroxidation, glutathione (GSH) content and some antioxidant enzymes activities in bone marrow cells of rats. Thirty-six rats were randomly classified into four groups. The first group was injected i.p. with ethanol and served as a control. The second group was treated with BCNU. The third group was given DMTU, while the fourth group was co-administered with DMTU prior to BCNU administration. BCNU treatment in a single dose of 30 mg/kg significantly decreased the normal counts of RBCs, WBCs and platelets as well as hemoglobin level. In addition, BCNU exhibited marked apoptotic effect associated with significant alterations in the oxidative cascade parameters. Treatment of animals with DMTU in a single dose of 500 mg/kg 1h before BCNU injection, followed by 125 mg/kg twice daily for 5 consecutive days significantly mitigated the induced changes in the hematological parameters. The induced alterations in the oxidant and antioxidant parameters as well as apoptosis were also improved. Conclusively, DMTU treatment exhibited marked chemopreventive effect against BCNU induced myelotoxicity; an effect which may be partially attributed to its inherently antioxidant potential. PMID- 21620923 TI - Modulation of genotoxicity and DNA repair by plant monoterpenes camphor, eucalyptol and thujone in Escherichia coli and mammalian cells. AB - The aim of this work was to examine the antigenotoxic potential of plant monoterpenes: camphor, eucalyptol and thujone in prokaryotic and eukaryotic cells and to elucidate their effect on DNA repair. We compared the effect of monoterpenes on spontaneous, UV- and 4NQO-induced mutagenesis in Escherichia coli K12 repair proficient, and MMR and NER deficient strains. Positive controls tannic acid and vanillin were included in bacterial tests. We also examined protective effect of monoterpenes against 4NQO-induced genotoxicity in Vero cell line by alkaline comet assay. The results obtained in repair proficient strain indicated antimutagenic potential of monoterpenes against UV- and 4NQO-induced mutagenesis, which was diminished with NER deficiency. Camphor and eucalyptol maintained UV-induced SOS response longer than in controls, while thujone decreased SOS response and reduced general protein synthesis and the growth rate. The three monoterpenes increased spontaneous and UV-induced recombination in recA730 and camphor additionally in recA(+) cells. Incubation of 4NQO-pretreated Vero cells with monoterpenes resulted in significant reduction of tail moment. However, higher concentrations of monoterpenes induced DNA strand breaks. Obtained results indicate that by making a small amount of DNA lesions camphor, eucalyptol and thujone can stimulate error-free DNA repair processes and act as bioantimutagens. PMID- 21620924 TI - Deoxynivalenol transport across the human placental barrier. AB - Deoxynivalenol (DON) is the most commonly detected mycotoxin contaminant of cereal crops and cereal based food products in temperate regions of the world. DON causes adverse health effects in animals, passes through to the foetus and causes foetal abnormalities in animals. Biomonitoring for DON has revealed frequent human exposure. This study reports on DON transfer across the human placenta. Firstly, in vitro studies with the BeWo b30 clone were used as a rapid screening model showing transfer of DON through a stable confluent cell monolayer. Five term placentas were then used to study DON transfer with the ex vivo dual perfusion model. The concentration of DON on the foetal side after 4h was about 21% of that on the maternal side at t=0. These results support the data from the BeWo monolayer model in respect to the transport rate of DON, and are consistent with our hypothesis of foetal exposure to DON during pregnancy. PMID- 21620925 TI - The effect of cooking on diphacinone residues related to human consumption of feral pig tissues. AB - We examined feral pig tissues to determine whether the potential hazard of consuming meat from pigs previously exposed to diphacinone rodenticide baits was reduced by cooking. Residue levels were measured in cooked and uncooked tissues of feral pigs exposed to sub-lethal quantities of the anticoagulant rodenticide. Pigs were provided large amounts of baits or untreated food to consume, then euthanized prior to the onset of symptoms indicative of rodenticide poisoning or sickness. For analysis, we grouped pigs into three levels of mean diphacinone consumption: 0, 3.5, and 7.4 mg/kg. None of the pigs displayed obvious signs of toxicity during the study period. The highest concentrations of diphacinone were found in liver tissue. Cooking had little effect on residual diphacinone concentrations. The hazards to humans and pets from meat from feral pigs that consumed the rodenticide diphacinone are not reduced by cooking; consumption of pig meat obtained from areas with active rodent control programs should be avoided. PMID- 21620926 TI - Fracture strength and fracture patterns of root filled teeth restored with direct resin restorations. AB - OBJECTIVE: To compare fracture characteristics of root-filled teeth with variable cavity design and resin composite restoration. METHODS: 80 extracted intact maxillary premolars were divided randomly into eight groups; (1) intact teeth; (2) unrestored MOD cavity; (3) unrestored MOD cavity plus endodontic access through the occlusal floor; (4) unrestored MOD plus endodontic access with axial walls removed; (5) MOD restored with resin composite; (6) MOD plus endodontic access, resin composite; (7) MOD plus extensive endodontic access, resin composite; (8) MOD plus extensive endodontic access, GIC core and resin composite. A ramped oblique load was applied to the buccal cusp in a servohydraulic testing machine. Fracture load and fracture patterns were recorded. Fracture loads were compared statistically using 1-way ANOVA, with Dunnett test for multiple comparisons. RESULTS: Unrestored teeth became progressively weaker with more extensive preparations. Endodontic access confined within the occlusal floor did not significantly affect strength compared to an MOD cavity. Loss of axial walls weakened teeth considerably [292+80N vs 747+130N for intact teeth]. Restoration increased the strength of prepared teeth particularly in teeth without axial walls. Teeth with a GIC core were not significantly weaker than intact teeth [560+167N]. Failures were mostly adhesive at the buccal interface, with the fracture propagating from the buccal line angle of the occlusal floor (MOD and MOD plus access groups) or of the proximal box (axial wall removed). CONCLUSIONS: Direct restorations increased fracture resistance of root filled teeth with extensive endodontic access. Both restored and unrestored teeth showed similar fracture patterns. PMID- 21620927 TI - Use of the checkerboard DNA-DNA hybridisation technique for in vivo detection of cariogenic microorganisms on metallic brackets, with or without use of an antimicrobial agent. AB - OBJECTIVE: Using checkerboard DNA-DNA hybridisation (CDDH) assay, this randomised clinical study evaluated the contamination of metallic brackets by four cariogenic bacterial strains (Streptococcus mutans, Streptococcus sobrinus, Lactobacillus casei and Lactobacillus acidophilus) and the efficacy of 0.12% chlorhexidine gluconate (CHX) mouthwashes in reducing bacterial contamination. METHODS: Thirty-nine 11-33-year-old patients under treatment with fixed orthodontic appliances were enrolled in the study and had 2 new metallic brackets bonded to premolars. Nineteen patients used a 0.12% CHX mouthwash (Periogard) and 20 patients used a placebo mouthwash (control) twice a week. After 30 days, the brackets were removed and samples were obtained for analysis by CDDH. Data were analysed statistically by the Kruskal-Wallis test (alpha=0.05) using the SAS software. RESULTS: S. mutans, S. sobrinus, L. casei and L. acidophilus were detected in 100% of the samples from both groups. However, brackets of the control group were more heavily contaminated by S. mutans and S. sobrinus (P<0.01). In the experimental group, although all counts decreased after rinsing with the chlorhexidine solution, there was significant difference only for S. mutans (P=0.03). CONCLUSIONS: The use of 0.12% chlorhexidine gluconate mouthwashes can be useful in clinical practice to reduce the levels of cariogenic microorganisms in patients under treatment with fixed orthodontic appliances. PMID- 21620928 TI - Revisiting intracellular calcium signaling semantics. AB - Cells use intracellular free calcium concentration changes for signaling. Signal encoding occurs through both spatial and temporal modulation of the free calcium concentration. The encoded message is detected by an ensemble of intracellular sensors forming the family of calcium-binding proteins (CaBPs) which must faithfully translate the message using a new syntax that is recognized by the cell. The cell is home to a significant although limited number of genes coding for proteins involved in the signal encoding and decoding processes. In a cell, only a subset of this ensemble of genes is expressed, leading to a genetic regulation of the calcium signal pathways. Calmodulin (CaM), the most ubiquitous expressed intracellular calcium-binding protein, plays a major role in calcium signal translation. Similar to a hub, it is central to a large and finely tuned network, receiving information, integrating it and dispatching the cognate response. In this review, we examine the different steps starting with an external stimulus up to a cellular response, with special emphasis on CaM and the mechanism by which it decodes calcium signals and translates it into exquisitely coordinated cellular events. By this means, we will revisit the calcium signaling semantics, hoping that we will ease communication between scientists dealing with calcium signals in different biological systems and different domains. PMID- 21620930 TI - Traffic optimization in railroad networks using an algorithm mimicking an amoeba like organism, Physarum plasmodium. AB - Traffic optimization of railroad networks was considered using an algorithm that was biologically inspired by an amoeba-like organism, plasmodium of the true slime mold, Physarum polycephalum. The organism developed a transportation network consisting of a tubular structure to transport protoplasm. It was reported that plasmodium can find the shortest path interconnecting multiple food sites during an adaptation process (Nakagaki et al., 2001. Biophys. Chem. 92, 47 52). By mimicking the adaptation process a path finding algorithm was developed by Tero et al. (2007). In this paper, the algorithm is newly modified for applications of traffic distribution optimization in transportation networks of infrastructure such as railroads under the constraint that the network topology is given. Application of the algorithm to a railroad in metropolitan Tokyo, Japan is demonstrated. The results are evaluated using three performance functions related to cost, traveling efficiency, and network weakness. The traffic distribution suggests that the modified Physarum algorithm balances the performances under a certain parameter range, indicating a biological process. PMID- 21620931 TI - Combining metabolic pathway analysis with Evolutionary Game Theory: explaining the occurrence of low-yield pathways by an analytic optimization approach. AB - Elementary-mode analysis is a powerful method for detecting all potential pathways in a metabolic network and computing the associated molar yields. Metabolic pathways can be interpreted as different strategies of organisms. Thus, methods from Evolutionary Game Theory can be employed. In Flux Balance Analysis (FBA), it is usually assumed that molar yields of relevant products (such as biomass or ATP) have been maximized during evolution. This has been questioned on game-theoretical grounds. In particular, in situations that can be characterized as a Prisoner's Dilemma, maximization of flux is not in line with maximization of yield. Under other conditions (that is, for other parameter values of maximal velocities), a Harmony game can result, where the above two maximization criteria give the same result. Here, we analyse the optimal situations under varying conditions. In particular, we consider the case where the cell can allocate a certain amount of protein on several enzymes in a varying distribution and model this by a linear programming problem in which not only the rates but also the maximal velocities are variable. It turns out that in the case of low or moderate synthesis costs for the enzymes of the high-yield pathway, maximizing pathway flux is in line with maximizing molar yield while in the case of high costs, it is not. This may explain the observation that many cells such as striated muscle cells, tumour cells, activated lymphocytes and several yeasts do not reallocate protein away from glycolytic enzymes towards TCA cycle and respiratory chain enzymes, in spite of the higher efficiency of respiration. This provides a straightforward explanation of the Warburg effect in tumour cells. PMID- 21620932 TI - The diffusion parameters of the extracellular space are altered in focal cortical dysplasias. AB - Most hypotheses concerning the mechanisms underlying seizure activity in focal cortical dysplasia (FCD) are based on alterations in synaptic transmission and glial dysfunction. However, neurons may also communicate by extrasynaptic transmission, which was recently found to affect epileptiform activity under experimental conditions and which is mediated by the diffusion of neuroactive substances in the extracellular space (ECS). The ECS diffusion parameters were therefore determined using the real-time iontophoretic method in human neocortical tissue samples obtained from surgically treated epileptic patients. The obtained values of the extracellular space volume fraction and tortuosity were then correlated with the histologicaly assessed type of cortical malformation (FCD type I or II). While the extracellular volume remained unchanged (FCD I) or larger (FCD II) than in normal/control tissue, tortuosity was significantly increased in both types of dysplasia, indicating the presence of additional diffusion barriers and compromised diffusion, which might be another factor contributing to the epileptogenicity of FCD. PMID- 21620934 TI - Time course of odorant- and trigeminal-induced activation in the human brain: an event-related functional magnetic resonance imaging study. AB - It is well known that most odorants stimulate the trigeminal system but the time course of the brain regions activated by these chemical stimulations remains poorly documented, especially regarding the trigeminal system. This functional magnetic resonance imaging (fMRI) study compares brain activations resulting from the contrast between two odorant conditions (one bimodal odor and one relatively pure olfactory stimulant) according to the duration of the stimulation (i.e. one inhalation, or three or six successive inhalations). The results show striking differences in the main brain regions activated according to these durations. The caudate nucleus and the orbitofrontal cortex are only involved in short-duration stimulations, and the posterior insular cortex and post-central gyrus (SI) are only activated by long duration stimulations. Different regions of the frontal, temporal and occipital lobe are activated depending on the duration but mainly during medium-duration stimulations. These results expand on the findings of previous studies and contribute to the description of temporal networks in trigeminal perception. PMID- 21620933 TI - Exposure of neurons to excitotoxic levels of glutamate induces cleavage of the RNA editing enzyme, adenosine deaminase acting on RNA 2, and loss of GLUR2 editing. AB - AMPA receptors are glutamate receptors that are tetramers of various combinations of GluR1-4 subunits. AMPA receptors containing GluR1, 3 and 4 are Ca2+ permeable, however, AMPA receptors containing even a single subunit of GluR2 are Ca2+ impermeable. Most AMPA receptors are Ca2+ impermeable due to the presence of GluR2. GluR2 confers special properties on AMPA receptors through the presence of arginine at the pore apex; other subunits (GluR1, 3, 4) contain glutamine at the pore apex and allow Ca2+ influx. Normally, an RNA editing step changes DNA encoded glutamine to arginine, introduces arginine in the GluR2 pore apex. GluR2 RNA editing is carried out by an RNA-dependent adenosine deaminase (ADAR2). Loss of GluR2 editing leads to the formation of highly excitotoxic AMPA channels [Mahajan and Ziff (2007) Mol Cell Neurosci 35:470-481] and is shown to contribute to loss of motor neurons in amyotrophic lateral sclerosis (ALS). Relatively higher levels of Ca2+-permeable AMPA receptors are found in motor neurons and this has been correlated with lower GluR2 mRNA levels. However, the reason for loss of GluR2 editing is not known. Here we show that exposure of neurons to excitotoxic levels of glutamate leads to specific cleavage of ADAR2 that leads to generation of unedited GluR2. We demonstrate that cleaved ADAR2 leads to a decrease or loss of GluR2 editing, which will further result in high Ca2+ influx and excitotoxic neuronal death. PMID- 21620929 TI - Mechanisms of specificity in neuronal activity-regulated gene transcription. AB - The brain is a highly adaptable organ that is capable of converting sensory information into changes in neuronal function. This plasticity allows behavior to be accommodated to the environment, providing an important evolutionary advantage. Neurons convert environmental stimuli into long-lasting changes in their physiology in part through the synaptic activity-regulated transcription of new gene products. Since the neurotransmitter-dependent regulation of Fos transcription was first discovered nearly 25 years ago, a wealth of studies have enriched our understanding of the molecular pathways that mediate activity regulated changes in gene transcription. These findings show that a broad range of signaling pathways and transcriptional regulators can be engaged by neuronal activity to sculpt complex programs of stimulus-regulated gene transcription. However, the shear scope of the transcriptional pathways engaged by neuronal activity raises the question of how specificity in the nature of the transcriptional response is achieved in order to encode physiologically relevant responses to divergent stimuli. Here we summarize the general paradigms by which neuronal activity regulates transcription while focusing on the molecular mechanisms that confer differential stimulus-, cell-type-, and developmental specificity upon activity-regulated programs of neuronal gene transcription. In addition, we preview some of the new technologies that will advance our future understanding of the mechanisms and consequences of activity-regulated gene transcription in the brain. PMID- 21620935 TI - Light and electron microscopic characterization of the evolution of cellular pathology in YAC128 Huntington's disease transgenic mice. AB - Huntington's disease (HD) is a progressive neurodegenerative disease caused by the insertion of an expanded polyglutamine sequence within the huntingtin protein. This mutation induces the formation of abnormal protein fragment aggregations and intra-nuclear neuronal inclusions in the brain. The present study aimed to produce a detailed longitudinal characterization of the neuronal pathology in the YAC128 transgenic mouse brain, to determine the similarity of this mouse model to other mouse models and the human condition in the spatial and temporal deposition pattern of the mutant protein fragments. Brain samples were taken from mice aged between 4 and 27 months of age, and assessed using S830 and GFAP immunohistochemistry, stereology and electron microscopy. Four month old mice did not exhibit intra-nuclear or extra-nuclear inclusions using the S830 antibody. Diffuse nuclear staining was present in the cortex, hippocampus and cerebellum from 6 months of age onwards. By 15 months of age, intra-nuclear inclusions were visible in most brain regions including nucleus accumbens, ventral striatum, lateral striatum, motor cortex, sensory cortex and cerebellum. The ventral striatum had a greater density of inclusions than the dorsal striatum. At 15 and 24 months of age, the mice showed increased reactive astrogliosis in the cortex, but no differences were found in the striatum. Necrotic cell death with vacuolation, uneven cell membrane and degenerated Golgi apparatus were detected ultrastructurally at 14 months of age, with some cells showing signs of apoptosis. By 26 months of age, most cells were degenerated in the transgenic animals, with lipofuscin granules being more abundant and larger in these mice than in their wildtype littermates. Our results demonstrate a progressive and widespread neuropathology in the YAC128 mice line that shares some similarity to the human condition. This article is part of a Special Issue entitled 'HD Transgenic Mouse'. PMID- 21620936 TI - Analysis of copy number variants in the cattle genome. AB - Copy number variation (CNV) is likely to be an important component of heritable variation in livestock. To characterise CNVs in cattle, we performed a genome wide survey to determine the number, location and gene content of these genomic features. A tiling oligonucleotide array with ~385,000 probes was used for comparative genomic hybridisation of both taurine and zebu cattle. Using a conservative set of calling criteria, a total of 51 CNV were detected that collectively spanned approximately half of one percent of the bovine genome. The size of the average CNV within each animal ranged from 213 kb up to 335 kb. Half of the CNV were detected in a single animal only, whilst the remainder was independently identified in multiple individuals. Analysis was performed to determine the gene content for each CNV region. This revealed that the majority of CNV (82%) spanned at least one gene, with a number of CNV containing genes which are known to control aspects of phenotypic variation in cattle. Whilst additional studies are required to determine the impact of individual CNV, this study confirmed them as an important class of genomic variation in cattle. PMID- 21620937 TI - Post-exposure therapy with recombinant human BuChE following percutaneous VX challenge in guinea-pigs. AB - Poisoning by nerve agents via the percutaneous (p.c.) route is an issue because the slow absorption of agent could result in poisoning which outlasts the protection provided by conventional pharmacological therapy. The bioscavenger approach is based on the concept of binding nerve agent in the bloodstream, thus preventing nerve agent from reaching the target tissues and inhibiting acetylcholinesterase activity. One bioscavenger that has been extensively studied is human butyrylcholinesterase (huBuChE). Protexia(r) is a pegylated form of recombinant huBuChE. We used a guinea-pig model of p.c. nerve agent poisoning, using an implanted telemetry system to collect physiological data. Guinea-pigs were poisoned with the nerve agent VX (0.74 mg/kg) (~2.5 * LD50). Two hours following VX exposure, Protexia (72 mg/kg) or saline control was administered intramuscularly. All guinea-pigs treated with Protexia (n=8) survived, compared to no survivors in a saline-treated control group (n=8). Survival following VX and Protexia treatment was associated with minimal incapacitation and observable signs of poisoning, and the mitigation or prevention of the detrimental physiological changes (e.g. seizure, bradycardia and hypothermia) observed in control animals. The opportunity for post-exposure treatment may have utility in both civilian and military scenarios, and this is a promising indication for the use of a bioscavenger. PMID- 21620938 TI - Hildebrand solubility parameter to predict drug release from hydroxypropyl methylcellulose gels. AB - An equation including the Hildebrand solubility parameter delta of the drugs is used for the first time to model drug release from hydroxypropyl methylcellulose (HPMC) gels: l nM = -21.578 + 2.102 delta-0.037 delta(2)+0.48 ln t + 1.028 ln C(i) (r(2) = 0.94 for a total of 286 cases). The experimentally determined release data of six drugs having different polarity (caffeine, theophylline, paracetamol, salicylic acid, naproxen and diclofenac) at several initial concentrations C(i) were included in the equation. In general, the amount of drug delivered is linear at the first 5-6h of the release profiles and the zero order constants K(o) increase as the solubility parameter of the drugs become larger. The Peppas exponential law M/M(infinity) = Kt(n) is applicable to larger fractional release, until 67-87% (48-51 h) for the less polar drugs (diclofenac and naproxen, lower delta values) and more than 80% (26-28 h) for the more polar drugs (higher delta values, theophylline, salicylic acid, caffeine and paracetamol). The Peppas release rate (lnK) shows a parabolic relationship with the drug solubility parameter. The diffusional exponent n varies between 0.40 and 0.58 indicating that drug release is mainly controlled by diffusion. An extended form of the Peppas equation is also tested for each drug including all the initial concentrations: lnM = a + b ln t + c ln C(i) (r(2) = 0.88-0.94). The logarithm of the octanol-water partition coefficients can also be used in combination with the drug concentrations. PMID- 21620939 TI - Regulation of sodium transport in the inner ear. AB - Na(+) concentrations in endolymph must be controlled to maintain hair cell function since the transduction channels of hair cells are cation-permeable, but not K(+)-selective. Flooding or fluctuations of the hair cell cytosol with Na(+) would be expected to lead to cellular dysfunction, hearing loss and vertigo. This review briefly describes cellular mechanisms known to be responsible for Na(+) homeostasis in each compartment of the inner ear, including the cochlea, saccule, semicircular canals and endolymphatic sac. The influx of Na(+) into endolymph of each of the organs is likely via passive diffusion, but these pathways have not yet been identified or characterized. Na(+) absorption is controlled by gate keeper channels in the apical (endolymphatic) membrane of the transporting cells. Highly Na(+)-selective epithelial sodium channels (ENaCs) control absorption by Reissner's membrane, saccular extramacular epithelium, semicircular canal duct epithelium and endolymphatic sac. ENaC activity is controlled by a number of signal pathways, but most notably by genomic regulation of channel numbers in the membrane via glucocorticoid signaling. Non-selective cation channels in the apical membrane of outer sulcus epithelial cells and vestibular transitional cells mediate Na(+) and parasensory K(+) absorption. The K(+)-mediated transduction current in hair cells is also accompanied by a Na(+) flux since the transduction channels are non-selective cation channels. Cation absorption by all of these cells is regulated by extracellular ATP via apical non-selective cation channels (P2X receptors). The heterogeneous population of epithelial cells in the endolymphatic sac is thought to have multiple absorptive pathways for Na(+) with regulatory pathways that include glucocorticoids and purinergic agonists. PMID- 21620941 TI - Physiological responses to chronic heat exposure in an aging non-human primate species, the gray mouse lemur (Microcebus murinus). AB - Epidemiological evidence related to increased death from hyperthermia suggests higher frailty in the elderly when exposed to high ambient temperatures. Despite the recent awareness of such public health problems, integrative studies investigating the effects of age on the physiological responses to heat wave thermal conditions remain scarce. Daily rhythmicity of core temperature (T(c)) and locomotor activity (LA), as well as parameters representative of energy balance and IGF-1 levels which are involved in the aging process and stress resistance, were monitored in a non-human primate species, the gray mouse lemur (Microcebus murinus). Adult and aged animals, acclimated to long days (LD) or short days (SD), were monitored during 8-day periods of exposure to 25 degrees C and 34 degrees C. Adult animals displayed efficient coping with heat exposure, regardless of the photoperiod. Hence, efficient responses resulted in decrease of energy intake, energy expenditure, IGF-1 levels and LA levels, promoting hyperthermia avoidance. Positive energy balance was maintained and water turnover did not change significantly after heat exposure. In contrast, while aged animals acclimated to LD responded similarly to adults, aged mouse lemurs acclimated to SD showed great difficulties coping with heat exposure. Indeed, in this group, normothermia impairment was associated with increased T(c) levels, alterations in daily rhythmicity, negative energy balance and increased IGF-1 levels. Impaired responses to heat exposure were seen in aged mouse lemurs acclimated to SD only. The main effects were observed during diurnal resting periods, suggesting decreased capacities with age to dissipate excess body heat. Taken together, these data highlight daily rhythmicity and IGF-1 pathway as main targets in the impaired response to heat exposure in the elderly. PMID- 21620940 TI - Plasticity of somatosensory inputs to the cochlear nucleus--implications for tinnitus. AB - This chapter reviews evidence for functional connections of the somatosensory and auditory systems at the very lowest levels of the nervous system. Neural inputs from the dosal root and trigeminal ganglia, as well as their brain stem nuclei, cuneate, gracillis and trigeminal, terminate in the cochlear nuclei. Terminations are primarily in the shell regions surrounding the cochlear nuclei but some terminals are found in the magnocellular regions of cochlear nucleus. The effects of stimulating these inputs on multisensory integration are shown as short and long-term, both suppressive and enhancing. Evidence that these projections are glutamatergic and are altered after cochlear damage is provided in the light of probable influences on the modulation and generation of tinnitus. PMID- 21620942 TI - The trypanothione system and the opportunities it offers to create drugs for the neglected kinetoplast diseases. AB - Parasitic trypanosomatids (Kinetoplastida) are the causative agents of devastating and hard-to-treat diseases such as African sleeping sickness, Chagas disease and various forms of Leishmaniasis. Altogether they affect > 30 Million patients, account for half a million fatalities p.a. and cause substantial economical problems in the Third World due to human morbidity and life stock losses. The design of efficacious and safe drugs is expected from inhibition of metabolic pathways that are unique and essential to the parasite and absent in the host. In this respect, the trypanothione system first detected in the insect pathogenic trypanosomatid Crithidia fasciculata qualified as an attractive drug target area. The existence of the system in pathogenic relatives was established by homology cloning and PCR. The vital importance of the system was verified in Trypanosoma brucei by dsRNA technology or knock-out in other trypanosomatids, respectively, and is explained by its pivotal role in the parasite's antioxidant defense and DNA synthesis. The key system component is the bis-glutathionyl derivative of spermidine, trypanothione. It is the proximal reductant of tryparedoxin which substitutes for thioredoxin-, glutaredoxin- and glutathione dependent reactions. Heterologous expression, functional characterization and crystallization of recombinant system components finally enable structure-based rational inhibitor design. PMID- 21620943 TI - Novel RasGRF1-derived Tat-fused peptides inhibiting Ras-dependent proliferation and migration in mouse and human cancer cells. AB - Mutations of RAS genes are critical events in the pathogenesis of different human tumors and Ras proteins represent a major clinical target for the development of specific inhibitors to use as anticancer agents. Here we present RasGRF1-derived peptides displaying both in vitro and in vivo Ras inhibitory properties. These peptides were designed on the basis of the down-sizing of dominant negative full length RasGRF1 mutants. The over-expression of these peptides can revert the phenotype of K-RAS transformed mouse fibroblasts to wild type, as monitored by several independent biological readouts, including Ras-GTP intracellular levels, ERK activity, morphology, proliferative potential and anchorage independent growth. Fusion of the RasGRF1-derived peptides with the Tat protein transduction domain allows their uptake into mammalian cells. Chemically synthesized Tat-fused peptides, reduced to as small as 30 residues on the basis of structural constraints, retain Ras inhibitory activity. These small peptides interfere in vitro with the GEF catalyzed nucleotide dissociation and exchange on Ras, reduce cell proliferation of K-RAS transformed mouse fibroblasts, and strongly reduce Ras-dependent IGF-I-induced migration and invasion of human bladder cancer cells. These results support the use of RasGRF1-derived peptides as model compounds for the development of Ras inhibitory anticancer agents. PMID- 21620944 TI - Computational model of EGFR and IGF1R pathways in lung cancer: a Systems Biology approach for Translational Oncology. AB - In this paper we propose a Systems Biology approach to understand the molecular biology of the Epidermal Growth Factor Receptor (EGFR, also known as ErbB1/HER1) and type 1 Insulin-like Growth Factor (IGF1R) pathways in non-small cell lung cancer (NSCLC). This approach, combined with Translational Oncology methodologies, is used to address the experimental evidence of a close relationship among EGFR and IGF1R protein expression, by immunohistochemistry (IHC) and gene amplification, by in situ hybridization (FISH) and the corresponding ability to develop a more aggressive behavior. We develop a detailed in silico model, based on ordinary differential equations, of the pathways and study the dynamic implications of receptor alterations on the time behavior of the MAPK cascade down to ERK, which in turn governs proliferation and cell migration. In addition, an extensive sensitivity analysis of the proposed model is carried out and a simplified model is proposed which allows us to infer a similar relationship among EGFR and IGF1R activities and disease outcome. PMID- 21620945 TI - BB14, a Nerve Growth Factor (NGF)-like peptide shown to be effective in reducing reactive astrogliosis and restoring synaptic homeostasis in a rat model of peripheral nerve injury. AB - Peptidomimetics hold a great promise as therapeutic agents for neurodegenerative disorders. We previously described a Nerve Growth Factor (NGF)-like peptide, now named BB14, which was found to act as a strong TrkA agonist and to be effective in the sciatic nerve injury model of neuropathic pain. In this report we present the effects of BB14 in reducing reactive astrocytosis and reverting neuroplastic changes of the glutamate/GABAergic circuitry in the lumbar spinal cord following spared nerve injury (SNI) of the sciatic nerve. Immunohistochemical analysis of spinal cord sections revealed that SNI was associated with increased microglial (Iba1) and astrocytic (GFAP) responses, indicative of reactive gliosis. These changes were paralleled by (i) decreased glial aminoacid transporters (GLT1 and GlyT1) and increased levels of (ii) neuronal glutamate transporter EAAC1, (iii) neuronal vesicular GABA transporter (vGAT) and (iv) the GABAergic neuron marker GAD65/67. A remarkable increase of the Glutamate/GABA ratio and the reduction of glutathione (GSH) levels were also indicative of modifications of glial function in neuroprotection. All these molecular changes were found to be linked to an alteration of endogenous NGF metabolism, as demonstrated by decreased levels of mature NGF, increase of proNGF and increased activity of NGF-degrading methallo proteinases (MMPs). Biochemical alterations and SNI-related neuropathic behavior, characterized by allodynia and hyperalgesia, were reversed by 7-days i.t. administration of the NGF-like peptide BB14, as well as by increasing endogenous NGF levels by i.t. infusion of GM6001, a MMPs inhibitor. All together, while confirming the correlation between reactive astrogliosis and perturbation of synaptic circuitry in the SNI model of peripheral nerve injury, these data strongly support the beneficial effect of BB14 in reducing reactive astrogliosis and restoring synaptic homeostasis under pathological conditions linked to alteration of NGF availability and signaling, thereby suggesting a potential role of BB14 as a therapeutic agent. PMID- 21620946 TI - Ion channels in key marine invertebrates; their diversity and potential for applications in biotechnology. AB - Of the intra-membrane proteins, the class that comprises voltage and ligand-gated ion channels represents the major substrate whereby signals pass between and within cells in all organisms. It has been presumed that vertebrate and particularly mammalian ion channels represent the apex of evolutionary complexity and diversity and much effort has been focused on understanding their function. However, the recent availability of cheap high throughput genome sequencing has massively broadened and deepened the quality of information across phylogeny and is radically changing this view. Here we review current knowledge on such channels in key marine invertebrates where physiological evidence is backed up by molecular sequences and expression/functional studies. As marine invertebrates represent a much greater range of phyla than terrestrial vertebrates and invertebrates together, we argue that these animals represent a highly divergent, though relatively underused source of channel novelty. As ion channels are exquisitely selective sensors for voltage and ligands, their potential and actual applications in biotechnology are manifold. PMID- 21620947 TI - Effects of pooling RNA from samples treated with different compounds for determining class specific biomarkers and processes in toxicogenomics. AB - Pooling of RNA samples is generally applied to obtain samples that represent the average signal of biological replicates of a single treatment. For toxicogenomics, pooling RNA of samples treated by different compounds could in the same way summarize these compounds to a single sample with average signals per class. In this study, we investigated the efficiency of such an approach to establish class specific differences in gene expression. As an example we took skin sensitizing compounds as one class and irritating compounds as another. A direct comparison was made to separately hybridized RNA samples. We observed that pooling RNA from compounds of a class substantially increased power to detect significantly regulated genes between classes because variability between pooled samples was much lower. Within pools the vast majority of genes maintained patterns of expression compared to the separately hybridized samples, especially in regulated genes. Both designs yielded appropriate biomarkers. Biomarkers selected from the pooled and separate design performed equally in classification of compounds to their class and relevant processes were found enriched in both designs. Consequently, pooling of RNA of different compound treated samples can be applied to determine class specific biomarkers and processes at much reduced cost and with limited loss of accuracy. PMID- 21620948 TI - Cellular glutathione in fatty liver in vitro models. AB - The range of non-alcoholic fatty liver disease (NAFLD) includes simple hepatic steatosis, the inflammatory non-alcoholic steatohepatitis (NASH), fibrosis and cirrhosis. The accumulation of specific lipids in hepatocytes has been associated with oxidative stress and progression of the disease. Elevated serum free fatty acids and hepatocyte lipotoxicity can be studied in an in vitro cellular model. For this purpose, we cultured the human liver cell line, HepG2/C3A, in medium supplemented with increasing amounts of oleic acid (C18:1) and evaluated oxidative stress by measuring the content of the cellular antioxidant, glutathione (GSH). We observed a dose-dependent steatosis, as determined by Nile Red staining, with concurrent increases of GSH; similar findings were also observed in cultured human hepatocytes. Cells cultured with palmitic acid (C16:0) or the combination oleic/palmitic acids (2:1 ratio) also exhibited a dose dependent increase of GSH; however palmitic-supplemented cultures did not sustain the GSH increase after 24h. We also detected an increase in the formation of lipid peroxides (LPO) indicating that the increase of GSH was a cellular mechanism that may be related to the high exposure of fatty acids. The results of this in vitro study suggest an antioxidant response against fat overloading and indicate potential differences in response to specific fatty acid-induced hepatic steatosis and associated lipotoxicity. PMID- 21620949 TI - Asymptotic distribution for epistatic tests in case-control studies. AB - We propose a statistical model for dissecting a multilocus genotypic value into its main (additive and dominant) effects and epistatic effects between different loci in a case-control association study. The model can discern four different kinds of epistasis, additive * additive, additive * dominant, dominant * additive, and dominant * dominant interactions. To test each kind of epistasis, a chi(2) test statistic was computed for a two by two contingency table derived from combined genotypes in both case and control groups. We derived an analytical approach for estimating the asymptotic distribution of the chi(2) test statistic for epistatic tests under the null hypothesis, with the result being consistent with that from Monte Carlo simulations. The new model was used to analyze a case control data set for candidate gene studies of stroke, leading to the identification of several significant interactions between causal SNPs on this disease. PMID- 21620950 TI - Methylation dynamics of IG-DMR and Gtl2-DMR during murine embryonic and placental development. AB - The Dlk1-Dio3 imprinted domain on mouse chromosome 12 contains IG-DMR and Gtl2 DMR, whose methylation patterns are established in the germline and after fertilization, respectively. In this study, we determine that acquisition of DNA methylation at the paternal allele of the Gtl2-DMR is initiated after the blastocyst stage and completed by embryonic day 6.5, and that Gtl2 (approved symbol: Meg3) is monoallelically expressed from the maternal allele as early as the blastocyst. Therefore, DNA methylation at the Gtl2-DMR is not a prerequisite for the imprinted expression of Gtl2, which may be involved in the control of proliferation and differentiation of cells during early gestation. We also reveal that a subregion of the IG-DMR exhibits tissue-specific differences in allelic methylation patterns. These results add to the growing body of knowledge elucidating the mechanism whereby parent-of-origin-dependent DNA methylation at the IG-DMR leads to the imprinted expression of the Dlk1-Dio3 cluster. PMID- 21620951 TI - BLBP-expression in astrocytes during experimental demyelination and in human multiple sclerosis lesions. AB - Several lines of evidence indicate that remyelination represents one of the most effective mechanisms to achieve axonal protection. For reasons that are not yet understood, this process is often incomplete or fails in multiple sclerosis (MS). Activated astrocytes appear to be able to boost or inhibit endogenous repair processes. A better understanding of remyelination in MS and possible reasons for its failure is needed. Using the well-established toxic demyelination cuprizone model, we created lesions with either robust or impaired endogenous remyelination capacity. Lesions were analyzed for mRNA expression levels by Affymetrix GeneChip(r) arrays. One finding was the predominance of immune and stress response factors in the group of genes which were classified as remyelination supporting factors. We further demonstrate that lesions with impaired remyelination capacity show weak expression of the radial-glia cell marker brain lipid binding protein (BLBP, also called B-FABP or FABP7). The expression of BLBP in activated astrocytes correlates with the presence of oligodendrocyte progenitor cells. BLBP-expressing astrocytes are also detected in experimental autoimmune encephalomyelitis during the remission phase. Furthermore, highest numbers of BLBP-expressing astrocytes were evident in lesions of early MS, whereas significantly less are present at the rim of (chronic)-active lesions from patients with long disease duration. Transfection experiments show that BLBP regulates growth factor expression in U87 astrocytoma cells. In conclusion, we provide evidence that expression of BLBP in activated astrocytes negatively correlates with disease duration and in parallel with remyelination failure. PMID- 21620952 TI - Reversal of imbalance between kynurenic acid and 3-hydroxykynurenine by antipsychotics in medication-naive and medication-free schizophrenic patients. AB - The association between the pro-inflammatory state of schizophrenia and increased tryptophan degradation into kynurenine has been reported. However, the relationship between metabolites from subdivisions of the kynurenine pathway, kynurenic acid and 3-hydroxykynurenine, remains unknown. The present study tested the relationship between these kynurenine metabolites in the plasma of medication naive (n=35) or medication-free (n=18) patients with schizophrenia at admission and following 6-week antipsychotic treatment compared to healthy controls (n=48). The plasma concentrations of kynurenic acid (nmol/l) were lower (difference=-8.44 (-13.22 to -3.65); p=0.001) and of 3-hydroxykynurenine (nmol/l) were higher (difference=11.24 (8.11-14.37); p<0.001) in the patients compared with the healthy controls. The kynurenic acid/kynurenine (difference=-2.75 (-5.115 to 0.336); p=0.026) and kynurenic acid/3-hydroxykynurenine (difference=-1.08 (-1.431 to -0.729); p<0.001) ratios were also lower in the patients. After the 6-week treatment, the patients' plasma kynurenic acid levels (difference=3.85 (-0.23 to 7.94); p=0.064) showed a trend towards an increase, whereas plasma 3 hydroxykynurenine levels (difference=22.41 (19.76-25.07); p<0.001) decreased. As a consequence, the kynurenic acid/3-hydroxykynurenine ratio (difference=-4.41 ( 5.51 to -3.3); p<0.001) increased. Higher initial plasma kynurenic acid levels on admission or increased kynurenic acid/kynurenine ratio after treatment were associated with reduction of clinical symptoms scores upon discharge although higher kynurenic acid/kynurenine on admission may induce higher positive symptoms score. In contrast, higher 3-hydroxykynurenine is associated with lower positive symptoms score. These results indicate that there is an imbalance in the kynurenine pathway in schizophrenia. The 6-week antipsychotic treatment may partially reverse the imbalance in kynurenine metabolism and that in turn induces clinical response. PMID- 21620953 TI - Intracerebroventricular administration of HIV-1 Tat induces brain cytokine and indoleamine 2,3-dioxygenase expression: a possible mechanism for AIDS comorbid depression. AB - Human immunodeficiency virus (HIV) remains a major public health concern despite a large education effort during the past 25 years. A persistent problem with HIV infection is the high comorbity rate of clinical depression. We previously established that increasing proinflammatory cytokines within the brain of mice induces sickness that can culminate in depressive-like behavior. Here we investigated the role of the HIV transactivator of transcription (Tat) protein in activation of brain cytokine signaling and subsequent induction of depressive like behavior in a murine model. Adult Balb/c mice were administered a single intracerebroventricular (ICV) injection of Tat (40 ng). Social investigation of a novel juvenile was measured at 2, 4, 8 and 24 h post-treatment. Mice treated with Tat did not display signs of sickness, as measured by either decreased social investigation or loss of body weight. At 24 h post-injection, mice were subjected to the forced swim test (FST). ICV administration of Tat to Balb/c mice increased immobility in the FST at 24 h post injection. A different strain of mice, C57BL/6J, responded similarly in the FST. Furthermore, adult C57BL/6J mice injected with Tat and tested in a two-bottle 1% sucrose preference test displayed reduced preference for sucrose during the 24 h post-injection period. Subsequently, brain tissues from Tat-treated and control C57BL/6J mice were collected at 4 and 24 h post injection. CNS tissue from Tat-treated mice had increased expression of IL-1beta, TNF-alpha, IL-6, and IDO mRNAs at 4 h post injection. These data demonstrate that a single exposure to Tat in the brain is sufficient to induce brain cytokine signaling that culminates in depressive-like behavior. The results reveal a potential role for Tat in the development of comorbid depression in HIV-infected individuals. PMID- 21620954 TI - Impact of perinatal exposure to equol enantiomers on reproductive development in rodents. AB - There is now considerable interest in the intestinally derived soy isoflavone metabolite, equol, which occurs in the enantiomeric forms, S-(-)equol and R (+)equol, both differing in biological actions. Little is known about effects of either enantiomer on reproductive development, yet such knowledge is fundamental because of the recent commercialization of S-(-)equol as a dietary supplement. S (-)equol and R-(+)equol were therefore investigated to determine their effects on reproductive development and fertility in the Sprague-Dawley rat. Neither enantiomer affected fertility, number of litters produced, number of pups per litter, number of male and female pups born, birth weight, anogenital distance, testicular descent or vaginal opening. Histological analysis showed no major abnormalities in ovary, testis, prostate or seminal vesicle tissue with dietary exposure to S-(-)equol or R-(+)equol, but both enantiomers triggered hyperplasia of uterine tissue. With R-(+)equol this stimulatory effect subsided after exposure was discontinued, but the effect of S-(-)equol was prolonged. PMID- 21620955 TI - Prenatal origin of obesity and their complications: Gestational diabetes, maternal overweight and the paradoxical effects of fetal growth restriction and macrosomia. AB - Pregestational (PGDM) and gestational (GDM) diabetes may be associated with a variety of fetal effects including increased rate of spontaneous abortions, intrauterine fetal death, congenital anomalies, neurodevelopmental problems and increased risk of perinatal complications. Additional problems of concern are fetal growth disturbances causing increased or decreased birth weight. Optimal control of maternal blood glucose is known to reduce these changes. Among the long lasting effects of these phenomena are a high rate of overweight and obesity at childhood and a high tendency to develop the "metabolic syndrome" characterized by hypertension, cardio-vascular complications and type 2 diabetes. Similarly, maternal overweight and obesity during pregnancy or excessive weight gain are also associated with increased obesity and complications in the offspring. Although there are different causes for fetal growth restriction (FGR) or for fetal excessive growth (macrosomis), paradoxically both are associated with the "metabolic syndrome" and its long term consequences. The exact mechanism(s) underlying these long term effects on growth are not fully elucidated, but they involve insulin resistance, fetal hyperleptinemia, hypothalamic changes and most probably epigenetic changes. Preventive measures to avoid the metabolic syndrome and its complications seem to be a tight dietary control and physical activity in the children born to obese or diabetic mothers or who had antenatal growth disturbances for other known or unknown reasons. PMID- 21620957 TI - ROS-activated p38 MAPK/ERK-Akt cascade plays a central role in palmitic acid stimulated hepatocyte proliferation. AB - In the past years, free fatty acids (FFAs) and obesity have been reported to play an important role in cancer development. Palmitic acid (PA) is the most prevalent saturated FFA in circulation. However, the mechanism underlying the effect of PA on cell proliferation is still to be elucidated. In this report, we, for the first time, investigate the signaling pathway in human normal hepatocytes (QZG) responsible for PA-induced proliferation. The results demonstrate that PA promotes cell cycle progression, accelerates cell proliferation, and induces a transient and sequential activation of a series of kinases. The employment of several inhibitors and antioxidants indicates that a ROS-induced stress-sensitive p38 MAPK/ERK-Akt cascade plays a critical role in the regulation of PA on cell cycle and cell proliferation. Moreover, PA dose and time dependently activates Nrf2 and this activation relies on ROS-induced stimulation of p38 MAPK/ERK-Akt signaling, demonstrating that Nrf2 activation may be associated with the regulation of PA on cell cycle transition and proliferation. In conclusion, our study elucidates the importance of PA metabolism on cell proliferation, and suggests that PA stimulates hepatocyte proliferation through activating the ROS p38 MAPK/ERK-Akt cascade which is intersected with the activation of Nrf2 and that the effect of ROS on signal transduction is in a dose- and time-dependent manner. All the above noted provide a new clue for the central role of ROS in cell proliferation and tumorigenesis. PMID- 21620958 TI - beta1-Integrin is up-regulated via Rac1-dependent reactive oxygen species as part of the hypertrophic cardiomyocyte response. AB - beta(1)-Integrin mediates cardiomyocyte growth and survival and its proper regulation is essential for the structural and functional integrity of the heart. beta(1)-Integrin expression is enhanced in hypertrophy, but the mechanism and significance of its up-regulation are unknown. Because reactive oxygen species (ROS) are important mediators of myocardial remodeling we examined their role in regulated beta(1)-integrin expression. Hypertrophy was induced in neonatal cardiomyocytes by endothelin-1 (ET-1), which activated the regulatory NADPH oxidase subunit Rac1, evoked ROS, and enhanced fetal gene expression and cardiomyocyte size. ET-1 also enhanced cell adhesion and FAK phosphorylation and inhibited oxidative stress-induced cardiomyocyte apoptosis. Further, ET-1 increased beta(1)-integrin mRNA and protein expression via Rac1-ROS-dependent MEK/ERK and EGF receptor-PI3K/Akt activation as shown by adenoviral dominant negative Rac1 or overexpression of copper/zinc-superoxide dismutase. The relevance of regulated beta(1)-integrin expression was examined in cardiomyocytes, in which targeting siRNA impeded the ET-1-induced beta(1) integrin up-regulation. In these cells, ET-1-induced cell adhesion, FAK phosphorylation, and hypertrophic response were significantly blunted, whereas its antiapoptotic effect was predominantly unchanged, suggesting at least partial dissociation of prohypertrophic and prosurvival signaling elicited by ET-1. In conclusion, beta(1)-integrin up-regulation in response to ET-1 is mediated via Rac1-ROS-dependent activation of prohypertrophic pathways and is mandatory for ET 1-induced FAK activation, cell adhesion, and hypertrophic response. PMID- 21620956 TI - Regulation of dorsal raphe nucleus function by serotonin autoreceptors: a behavioral perspective. AB - Neurotransmission by serotonin (5-HT) is tightly regulated by several autoreceptors that fine-tune serotonergic neurotransmission through negative feedback inhibition at the cell bodies (predominantly 5-HT(1A)) or at the axon terminals (predominantly 5-HT(1B)); however, more subtle roles for 5-HT(1D) and 5 HT(2B) autoreceptors have also been detected. This review provides an overview of 5-HT autoreceptors, focusing on their contribution in animal behavioral models of stress and emotion. Experiments targeting 5-HT autoreceptors in awake, behaving animals have generally shown that increasing autoreceptor feedback is anxiolytic and rewarding, while enhanced 5-HT function is aversive and anxiogenic; however, the role of serotonergic activity in behavioral models of helplessness is more complex. The prevailing model suggests that 5-HT autoreceptors become desensitized in response to stress exposure and antidepressant administration, two seemingly opposite manipulations. Thus there are still unresolved questions regarding the role of these receptors-and serotonin in general-in normal and pathological states. PMID- 21620959 TI - Leishmania-macrophage interactions: insights into the redox biology. AB - Leishmaniasis is a neglected tropical disease that affects about 350 million individuals worldwide. The protozoan parasite has a relatively simple life cycle with two principal stages: the flagellated mobile promastigote living in the gut of the sandfly vector and the intracellular amastigote within phagolysosomal vesicles of the vertebrate host macrophage. This review presents a state-of-the art overview of the redox biology at the parasite-macrophage interface. Although Leishmania species are susceptible in vitro to exogenous superoxide radical, hydrogen peroxide, nitric oxide, and peroxynitrite, they manage to survive the endogenous oxidative burst during phagocytosis and the subsequent elevated nitric oxide production in the macrophage. The parasite adopts various defense mechanisms to cope with oxidative stress: the lipophosphoglycan membrane decreases superoxide radical production by inhibiting NADPH oxidase assembly and the parasite also protects itself by expressing antioxidant enzymes and proteins. Some of these enzymes could be considered potential drug targets because they are not expressed in mammals. In respect to antileishmanial therapy, the effects of current drugs on parasite-macrophage redox biology and its involvement in the development of drug resistance and treatment failure are presented. PMID- 21620960 TI - Akt signalling in health and disease. AB - Akt (also known as protein kinase B or PKB) comprises three closely related isoforms Akt1, Akt2 and Akt3 (or PKBalpha/beta/gamma respectively). We have a very good understanding of the mechanisms by which Akt isoforms are activated by growth factors and other extracellular stimuli as well as by oncogenic mutations in key upstream regulatory proteins including Ras, PI3-kinase subunits and PTEN. There are also an ever increasing number of Akt substrates being identified that play a role in the regulation of the diverse array of biological effects of activated Akt; this includes the regulation of cell proliferation, survival and metabolism. Dysregulation of Akt leads to diseases of major unmet medical need such as cancer, diabetes, cardiovascular and neurological diseases. As a result there has been substantial investment in the development of small molecular Akt inhibitors that act competitively with ATP or phospholipid binding, or allosterically. In this review we will briefly discuss our current understanding of how Akt isoforms are regulated, the substrate proteins they phosphorylate and how this integrates with the role of Akt in disease. We will furthermore discuss the types of Akt inhibitors that have been developed and are in clinical trials for human cancer, as well as speculate on potential on-target toxicities, such as disturbances of heart and vascular function, metabolism, memory and mood, which should be monitored very carefully during clinical trial. PMID- 21620961 TI - (R)-FTY720 methyl ether is a specific sphingosine kinase 2 inhibitor: Effect on sphingosine kinase 2 expression in HEK 293 cells and actin rearrangement and survival of MCF-7 breast cancer cells. AB - Sphingosine kinase 2 (SK2) catalyses the conversion of sphingosine to the bioactive lipid sphingosine 1-phosphate (S1P). We report here, the stereospecific synthesis of an analogue of FTY720 called (R)-FTY720-OMe, which we show is a competitive inhibitor of SK2. (R)-FTY720-OMe failed to inhibit sphingosine kinase 1 activity, thereby demonstrating specificity for SK2. Prolonged treatment of HEK 293 cells with (R)-FTY720-OMe also induced a reduction in SK2 expression. In addition, (R)-FTY720-OMe inhibited DNA synthesis and prevented S1P-stimulated rearrangement of actin in MCF-7 breast cancer cells. These findings demonstrate that SK2 functions as a pro-survival protein and is involved in promoting actin rearrangement into membrane ruffles/lamellipodia in response to S1P in MCF-7 breast cancer cells. PMID- 21620962 TI - Cytotoxic T lymphocyte antigen-2 alpha induces apoptosis of murine T-lymphoma cells and cardiac fibroblasts and is regulated by cAMP/PKA. AB - The mechanism of cAMP-promoted apoptosis is not well defined. In wild-type (WT) murine S49 lymphoma cells, cAMP promotes apoptosis in a protein kinase A (PKA) dependent manner. We find that treatment of WT S49 cells with 8-CPT-cAMP prominently increases the expression (as determined by DNA microarray analysis, real-time PCR and immunblotting) of cytotoxic T lymphocyte antigen-2alpha (CTLA 2alpha), a cathepsin L-like cysteine protease inhibitor. By contrast, CTLA-2alpha expression is only slightly increased by 8-CPT-cAMP treatment of D-S49 cells, which lack cAMP/PKA-promoted apoptosis. Raising endogenous cAMP (by use of forskolin or inhibition of phosphodiesterase [PDE] 4) or a PKA-selective, but not an Epac-selective, cAMP analogue, increases CTLA-2alpha mRNA expression; PKA, and not Epac, thus mediates the increase in CTLA-2alpha expression. An adenoviral CLTA-2alpha (Ad-CTLA-2alpha) construct induces apoptosis and enhances cAMP promoted apoptosis in WT S49 cells but such cells do not have an increase in cathepsin L activity nor does a cathepsin L inhibitor alter cAMP-promoted apoptosis. 8-CPT-cAMP also increases CTLA-2alpha expression and induces apoptosis in murine cardiac fibroblasts; knockdown of CTLA-2alpha expression by siRNA blocks 8-CPT-cAMP-promoted apoptosis. Thus, cAMP increases CTLA-2alpha expression in murine lymphoma and cardiac fibroblasts and this increase in CTLA-2alpha contributes to cAMP/PKA-promoted apoptosis by mechanisms that are independent of the ability of CTLA-2alpha to inhibit cathepsin L. PMID- 21620963 TI - Erythropoietin protects retinal pigment epithelial cells against the increase of permeability induced by diabetic conditions: essential role of JAK2/ PI3K signaling. AB - The outer blood-retinal barrier is formed by retinal pigment epithelial (RPE) cells and its disruption significantly contributes to the development of diabetic macular edema (DME). The aim of the study was to explore whether erythropoietin (Epo) has beneficial effects on the barrier function of human RPE cells and the main downstream pathways involved. ARPE-19 cells were cultured in standard conditions and under conditions leading to the disruption of the monolayer [25 mmol/L D-glucose plus IL-1beta (10 ng/mL)]. Epo (200 mU/mL/day) was added during the last 2 days of the experiment. The experiments were repeated in the presence of an Epo neutralizing antibody and specific inhibitors of JAK2 and PI3K (AG490 and LY294002, respectively). Permeability was evaluated by fluorescein isothiocyanate dextran (70 kDa) movements. Distribution of tight junction proteins was examined by immunofluorescence. Changes in cytosolic Ca(2+) induced by Epo were also measured. Epo treatment was able to prevent but not to restore the increase of permeability induced by high glucose plus IL-1beta. The protective effect of Epo on RPE barrier function was completely blocked by AG490 and almost completely abolished by LY294002. In addition, Epo was able to increase cytosolic Ca(2+) with dependence on extracellular calcium influx and this effect was blocked by either JAK2 or PI3K inhibition. We conclude that RPE disruption induced by high glucose plus IL-1beta is prevented by Epo through the downstream signaling of JAK2 and PI3K/AKT pathways. PMID- 21620964 TI - An IkappaBalpha phosphorylation inhibitor induces heme oxygenase-1(HO-1) expression through the activation of reactive oxygen species (ROS)-Nrf2-ARE signaling and ROS-PI3K/Akt signaling in an NF-kappaB-independent mechanism. AB - Reactive oxygen species (ROS) are important signaling molecules in cells. Excessive ROS induce expression of inflammatory mediators, such as iNOS and COX2. Antioxidant enzymes, such as, heme oxygenase-1 (HO-1), tightly regulate ROS levels within cells. Here, we show that Bay 11-7082 (Bay) increased HO-1 mRNA and protein expression in human colon cancer HT29 cells. Bay induced translocation of NF-E2-related factor 2 (Nrf2) into nuclei and increased the binding activity of the antioxidant response element (ARE). In addition, PI3K/Akt inhibitor (LY294002) blocked Bay-induced HO-1 expression. Pretreatment with anti-oxidants (N-acetylcysteine (NAC) or glutathione) significantly reduced Bay-induced HO-1 mRNA/protein expression, nuclear translocation of Nrf2 and phosphorylation of Akt. However, PI3K/Akt signaling was independent of Bay-induced Nrf2 translocation and ARE binding activity. Furthermore, other NF-kappaB inhibitors, such as pyrrolidine dithiocarbamate (PDTC) and MG132, also increased HO-1 mRNA and protein expression. However, although overexpression of dominant negative inhibitory kappaB (IkappaB) reduced NF-kappaB-driven transcriptional activity, IkappaB overexpression did not increase HO-1 expression. Taken together, our results suggest that in human colon cancer HT29 cells, Bay induces HO-1 expression by increasing ROS production in an Nrf2-ARE and PI3K dependent manner, but Bay acts independently of NF-kappaB. PMID- 21620965 TI - Conformational conversion of PDE5 by incubation with sildenafil or metal ion is accompanied by stimulation of allosteric cGMP binding. AB - Phosphodiesterase-5 (PDE5) is a dimer containing a cGMP-specific catalytic domain and an allosteric cGMP-binding subdomain (GAF A) on each subunit. PDE5 exhibits three conformational forms that can be separated by Native PAGE and are denoted as Bands 1, 2, and 3 in decreasing order of mobility. A preparation comprised mainly of Band 2 PDE5 was partially converted to Band 3 PDE5 by 1h incubation with cGMP or the PDE5-specific inhibitors sildenafil, vardenafil, or tadalafil, but not with cAMP, milrinone (PDE3-specific), or rolipram (PDE4-specific). Band 2 PDE5 was converted almost entirely to Band 3 PDE5 by overnight incubation with sildenafil at 30 degrees C. This time-dependent conversion was accompanied by a 7 fold increase in allosteric cGMP-binding activity, suggesting that Band 3 PDE5 is a much more active form than Band 2 PDE5 for allosteric cGMP binding. Conversion of Band 2 PDE5 to Band 3 PDE5 occurred faster by pre-incubation with cGMP, which binds to both the allosteric and catalytic sites of PDE5, than with catalytic site-specific sildenafil. Overnight incubation of a Band 2/Band 3 PDE5 mixture with EDTA caused time-dependent conversion to Band 1 PDE5 (apoenzyme), and this conversion was accompanied by a 50% loss in cGMP-binding activity. After incubation with EDTA, addition of Mn(++) or Mg(++) caused reversion of Band 1 to a Band 2/Band 3 PDE5 mixture in which Band 3 PDE5 predominated. This reversion was accompanied by a 3-fold increase in allosteric cGMP-binding activity. The combination of results implied that physiological conversion of Band 2 to Band 3 PDE5 by cGMP and/or divalent metal ion occupancy of the catalytic domain would increase allosteric cGMP binding to the enzyme. This conversion would produce a greater negative feedback effect on cGMP action by increasing sequestration of cGMP at the allosteric cGMP-binding site of PDE5 and by increasing cGMP degradation at the catalytic site of the enzyme. This conversion would also increase PDE5 inhibitor binding to the enzyme. PMID- 21620967 TI - Physical and catalytic properties of a peroxidase derived from cytochrome c. AB - Except for its redox properties, cytochrome c is an inert protein. However, dissociation of the bond between methionine-80 and the heme iron converts the cytochrome into a peroxidase. Dissociation is accomplished by subjecting the cytochrome to various conditions, including proteolysis and hydrogen peroxide (H(2)O(2))-mediated oxidation. In affected cells of various neurological diseases, including Parkinson's disease, cytochrome c is released from the mitochondrial membrane and enters the cytosol. In the cytosol cytochrome c is exposed to cellular proteases and to H(2)O(2) produced by dysfunctional mitochondria and activated microglial cells. These could promote the formation of the peroxidase form of cytochrome c. In this study we investigated the catalytic and cytolytic properties of the peroxidase form of cytochrome c. These properties are qualitatively similar to those of other heme-containing peroxidases. Dopamine as well as sulfhydryl group-containing metabolites, including reduced glutathione and coenzyme A, are readily oxidized in the presence of H(2)O(2). This peroxidase also has cytolytic properties similar to myeloperoxidase, lactoperoxidase, and horseradish peroxidase. Cytolysis is inhibited by various reducing agents, including dopamine. Our data show that the peroxidase form of cytochrome c has catalytic and cytolytic properties that could account for at least some of the damage that leads to neuronal death in the parkinsonian brain. PMID- 21620966 TI - Altered expression and function of regulator of G-protein signaling-17 (RGS17) in hepatocellular carcinoma. AB - Guanine nucleotide regulatory proteins (G-proteins) are central to normal hepatocyte function and are implicated in hepatic disease initiation and progression. Regulators of G-protein signaling (RGS) are critical to defining G protein-dependent signal fidelity, yet the role of RGS proteins in the liver is poorly defined. The aims of this study were to determine RGS17 expression in normal and transformed hepatic tissue and cells, and address the function of RGS17 in hepatic tumorgenicity. RGS17 expression was determined in human and rat HCC tissue and cell lines. Molecular approaches were used to alter RGS17 expression in HCC cells, effects on cell function measured, and RGS17 association with specific Galpha-subunits determined. Using these approaches RGS17 mRNA, but not protein, was detectable in human and rat HCC tissue and cells. Conversely, RGS17 mRNA was not detected in normal tissue, isolated hepatocytes, or non tumorigenic hepatic cells. Subsequent studies using transfected cells demonstrated that RGS17 proteins were not post-translationally modified in HCC cells, and RGS17 expression is governed by protein degradation and not via miRNAs. Notwithstanding inherently low RGS17 protein levels, altering RGS17 expression profoundly affected HCC cell mitogenesis and migration. Analysis of RGS17-G-protein interaction demonstrated RGS17 associates with both Gialpha- and Gqalpha-subunits in HCC cells of human and rat origin. In conclusion, these data demonstrate that, despite difficulties in measuring endogenous RGS protein expression, RGS17 is differentially expressed in HCC and plays a central role in regulating transformed hepatocyte tumorgenicity. PMID- 21620968 TI - Sonosensitive dioleoylphosphatidylethanolamine-containing liposomes with prolonged blood circulation time of doxorubicin. AB - Ultrasound sensitive (sonosensitive) liposomes represent a drug delivery system designed for releasing a drug load upon exposure to ultrasound (US). Inclusion of dioleoylphosphatidylethanolamine (DOPE) in liposome membranes was previously shown to induce sonosensitivity. Long blood circulation time of the liposomal drug is required for high tumour uptake and efficient US-mediated drug delivery. In this study, blood pharmacokinetics of DOPE-based liposomal doxorubicin (DXR) were evaluated in non-tumoured mice. A markedly faster blood clearance of DXR was observed for DOPE-rich liposomes compared to Caelyx(r) (standard liposomal DXR). Subsequently, liposome membrane composition was altered to improve drug retention in the bloodstream, whilst maintaining sonosensitivity. Formulations with reduced blood clearance of DXR were obtained by reducing the content of DOPE from 62 to 32 or 25 mol%. These formulations showed long blood circulation time, as approximately 20% of the administered DXR dose was present in the bloodstream 24 h after intravenous injection. The reduction in liposomal DOPE content did not significantly reduce US-mediated DXR release in vitro, indicating that DOPE is a potent modulator of sonosensitivity. The novel liposome formulations, containing moderate amounts of DOPE, displayed similar blood pharmacokinetic profiles as standard liposomal DXR, but a markedly improved sonosensitivity. PMID- 21620969 TI - Ex vivo decrease in uranium diffusion through intact and excoriated pig ear skin by a calixarene nanoemulsion. AB - Cutaneous contamination by radionuclides is a major concern in the nuclear industry. In case of skin exposure to uranium, no efficient emergency treatment is available to remove the actinide from the skin. For this purpose, we developed a nanoemulsion containing calixarene molecules displaying good chelating properties towards uranium. In this paper, we describe the ability of this formulation to trap uranium and limit its transfer from the cutaneous contaminated site into the blood. Uranium percutaneous diffusion kinetics was assessed with Franz cells over 24 h through intact and excoriated pig ear skin biopsies, after or without application of the nanoemulsion. Uranium distribution in the skin layers was analysed by SIMS microscopy. The results showed that prompt application of the calixarene nanoemulsion allows a 94% and 98% reduction of the amount of uranium diffused respectively through intact and excoriated skin. The formulation is still efficient in case of delayed application up to 30 minutes since the 24 h-uranium transfer through excoriated skin is reduced by 71%. Besides, no accumulation of uranium or uranium-calixarene chelate was observed in the different skin layers. In conclusion, this study demonstrated the efficiency of the calixarene nanoemulsion, which can be regarded as a promising treatment for uranium cutaneous contamination. PMID- 21620970 TI - Effects of PEG size on structure, function and stability of PEGylated BSA. AB - The effects of PEGylation on the structural, thermal and functional stability of bovine serum albumin (BSA) were investigated using BSA and 6 linear mono PEGylated BSA compounds. The secondary and tertiary structure of BSA measured by circular dichroism (CD) was independent of PEGylation. In contrast, the thermal stability of BSA was affected by PEGylation. The apparent unfolding temperature T(max) measured by differential scanning calorimetry (DSC) decreased with PEGylation, whereas the temperature of aggregation, T(agg), measured by dynamic light scattering (DLS) increased with PEGylation. The unfolding temperature and the temperature of aggregation were both independent of the molecular weight of the PEG chain. Possible functional changes of BSA after PEGylation were measured by Isothermal Titration Calorimetry (ITC), where the binding of sodium dodecyl sulphate (SDS) to BSA and PEGylated BSA was analysed. At 25 degrees C, two distinct classes of binding sites (high affinity and low affinity) for BSA and one class of binding site (low affinity) for PEGylated BSA were identified. The binding isotherm was modelled assuming independence and thermodynamic equivalence of the sites within each class. From the present biophysical characterisation, it is concluded that after PEGylation BSA appears to be unaffected structurally (secondary and tertiary structure), slightly destabilised thermally (unfolding temperature), stabilised kinetically (temperature of aggregation) and has an altered functionality (binding profile). These biophysical characteristics are all independent of the molecular weight of the attached polymer chain. PMID- 21620971 TI - Expression of hexose-6-phosphate dehydrogenase in rat tissues. AB - Hexose-6-phosphate dehydrogenase (H6PD) is the main NADPH generating enzyme in the lumen of the endoplasmic reticulum. H6PD is regarded as an ancillary enzyme in prereceptorial glucocorticoid activation and probably acts as a nutrient sensor and as a prosurvival factor. H6PD expression was determined in a variety of rat and human tissues by detecting mRNA and protein levels, and by measuring its dehydrogenase and lactonase activities. It was found that H6PD was present in all investigated tissues; both expression and activity remained within an order of magnitude. Correlation was found between the dehydrogenase activity and protein or mRNA levels. The results confirmed the supposed housekeeping feature of the enzyme. PMID- 21620972 TI - Thymic function, anti-thymocytes globulins, and cancer after renal transplantation. AB - BACKGROUND: Prolonged CD4 T cell lymphopenia after polyclonal antithymocyte globulins (ATG) is associated with an increased rate of cancers. Here, we examined whether pre-transplant thymic function estimated by TREC levels is predictive of cancer occurrence following ATG treatment. PATIENTS AND METHODS: The impact of TREC on cancer occurrence was analyzed in 115 consecutive incident renal transplant recipients having received ATG. RESULTS: Mean follow-up was 7.5+/-2.6years. After ATG induction, patients with the lowest pre-transplant TREC values had lower post-transplant CD4(+) and CD4(+) CD45RA(+) CD45RO(-) T cell counts, and a higher frequency of T cells with a regulatory phenotype (CD127(+)CD4(+)CD25(+)Foxp3(+)). Log-transformed pre-transplant TREC values were significantly lower in patients who developed cancer after transplantation (p<0.0001). The cumulative incidence of cancer was higher in patients having the lowest pre-transplant TREC values (T1 [low]: 47.4%, T2 [medium]: 12.5%, and T3 [high]: 2.7%; p<0.0001). In multivariate analysis, pre-transplant TREC value was the only predictive factor of cancer (HR, 0.39; 95% CI, 0.16 to 0.97, for one log (TREC/10(6) PBMC); p=0.046). CONCLUSIONS: Pre-transplant thymic function is associated with an increased rate of post-transplant cancer in patients having received ATG. Omitting ATG in recipients with low pre-transplant TREC values should be considered. PMID- 21620973 TI - Investigating a macromolecular complex: the toolkit of methods. AB - Structural biologists studying macromolecular complexes spend considerable effort doing strictly "non-structural" work: investigating the physiological relevance and biochemical properties of a complex, preparing homogeneous samples for structural analysis, and experimentally validating structure-based hypotheses regarding function or mechanism. Familiarity with the diverse perspectives and techniques available for studying complexes helps in the critical assessment of non-structural data, expedites the pre-structural characterization of a complex and facilitates the investigation of function. Here we survey the approaches and techniques used to study macromolecular complexes from various viewpoints, including genetics, cell and molecular biology, biochemistry/biophysics, structural biology, and systems biology/bioinformatics. The aim of this overview is to heighten awareness of the diversity of perspectives and experimental tools available for investigating complexes and of their usefulness for the structural biologist. PMID- 21620974 TI - Expression of immune-related genes in rainbow trout (Oncorhynchus mykiss) induced by probiotic bacteria during Lactococcus garvieae infection. AB - The aim of the present study was to investigate the effect of lactic acid bacteria (LAB) on the control of lactococcosis as well as to assess the impact of probiotics on the expression of immune-related genes in the head kidney and intestine of rainbow trout (Oncorhynchus mykiss). Lactobacillus plantarum, Lactococcus lactis and Leuconostoc mesenteroides, were administered orally at 106 CFU g-1 feed to fish for 36 days. Twenty-one days after the start of the feeding period, fish were challenged with Lactococcus garvieae. Only the fish fed the diet containing Lb. plantarum showed significantly (P < 0.05) improved protection against L. garvieae compared to the control. Subsequently, real-time PCR was employed to determine the mRNA levels of IL-1beta, IL-8, IL-10 and TNF-alpha in the head kidney, and IL-8, Tlr5 and IgT in the intestine of the control and Lb. plantarum groups. IL-1beta, IL-10 and TNF-alpha gene expression were significantly up-regulated by Lb. plantarum. Moreover, the mRNA levels of IL-10, IL-8 and IgT were significantly higher in the Lb. plantarum group after L. garvieae infection, suggesting that Lb. plantarum can stimulate the immune response of rainbow trout. PCR-DGGE revealed no detectable levels of the probiotics or the pathogen present on the distal intestinal mucosa. These findings demonstrate that direct probiotic-host interactions with the intestine are not always necessary to induce host stimulatory responses which ultimately enhance disease resistance. Furthermore, as L. garvieae did not colonise the intestinal tract, and therefore likely did not infect via this route, the antagonistic properties of the probiotic candidate towards L. garvieae were likely of little influence in mediating the improved disease resistance which could be attributed to the elevated immunological response. PMID- 21620975 TI - Two types of calmodulin play different roles in Pacific white shrimp (Litopenaeus vannamei) defenses against Vibrio parahaemolyticus and WSSV infection. AB - Calmodulin (CaM) plays an important role in calcium-dependent signal transduction pathways. In the present study, two alternative splicing isoforms of CaM (named LvCaM-A and LvCaM-B) cDNA were cloned from the Pacific white shrimp, Litopenaeus vannamei. LvCaM-A was of 1101 bp, including a 5'-terminal untranslated region (UTR) of 70 bp, a 3'-terminal UTR of 581 bp and an open reading frame (ORF) of 450 bp encoding a polypeptide of 149 amino acids with a calculated molecular weight (Mw) of 17 kDa and pI of 4.41. LvCaM-B was 689 bp, including a same 5'-UTR of 70 bp, a 3'-terminal UTR of 109 bp and an ORF of 510 bp encoding a polypeptide of 169 amino acids with a calculated Mw of 19 kDa and pI of 4.36. Both LvCaM-A and LvCaM-B contained 4 conservative EF-hand motifs. Quantitative real-time reverse transcription PCR analysis revealed LvCaM-A to be expressed in most shrimp tissues, with the predominant expression in nerve and the weakest expression in heart. However, LvCaM-B expression level was much weaker than those of LvCaM-A in all the tested tissues with main expression in hepatopancreas. The expression of LvCaM-A and LvCaM-B after challenge with Vibrio parahaemolyticus and WSSV were tested in hemocytes, hepatopancreas and nerve. The results indicated that LvCaM-A and LvCaM-B transcripts could be significantly induced in hemocytes and hepatopancreas respectively by injection with V. parahaemolyticus. The highest expression of LvCaM-A was in the hemocytes with 2.3 times (at 48 h) greater expression than in the control (p < 0.05). However, sharp down-regulation of both LvCaM-A and LvCaM-B were detected in nerve after Vibrio- and WSSV injection (p < 0.05). These results suggested that CaM might play an important role in shrimp's defense against pathogenic infection. PMID- 21620976 TI - Quantitative gene expression and in situ localization of scygonadin potentially associated with reproductive immunity in tissues of male and female mud crabs, Scylla paramamosain. AB - Scygonadin (Scy) is an important antimicrobial peptide which was first isolated from the seminal plasma of Scylla serrata (now renamed as Scylla paramamosain). Elucidation of the Scy expression pattern in tissues will help in understanding its potential function associated with the reproductive immunity. In our study, Scy mRNA transcripts and its protein were found widely distributed in mature male and female crabs. Scy mRNA transcripts were significantly demonstrated in the ejaculatory duct and hemocytes of males but were much less expressed in the other tissues tested. In addition, Scy mRNA transcripts were discerned in a number of cells in the glandular epithelium of the inner wall and in the secretion inside the ejaculatory duct using the in situ hybridization method. In females, Scy mRNA transcripts were obviously demonstrated in the hemocytes and gills but weakly detected in other tissues tested. The copy number of scygonadin mRNA transcripts in the ejaculatory duct of males was greatly higher than those in other tissues, in particular, was over 60,000 fold that in the hemocytes of females. Using immunohistochemistry, the Scy protein was found at higher levels in male tissues than in female ones, particularly in the reproductive duct of males. It was also interesting to note that Scy gene expression was not significantly induced with lipopolysaccharide challenge. However, it was highly expressed in the ejaculatory duct and the seminal vesicle of pre-copulatory males and in the spermathecae of post-copulatory females under mating conditions. The results suggested that Scy, as an important antimicrobial component, probably performed more functions in males, and was likely to be involved in a function associated with crab fertilization and reproduction in both males and females during mating. PMID- 21620977 TI - MEG beamforming using Bayesian PCA for adaptive data covariance matrix regularization. AB - Beamformers are a commonly used method for doing source localization from magnetoencephalography (MEG) data. A key ingredient in a beamformer is the estimation of the data covariance matrix. When the noise levels are high, or when there is only a small amount of data available, the data covariance matrix is estimated poorly and the signal-to-noise ratio (SNR) of the beamformer output degrades. One solution to this is to use regularization whereby the diagonal of the covariance matrix is amplified by a pre-specified amount. However, this provides improvements at the expense of a loss in spatial resolution, and the parameter controlling the amount of regularization must be chosen subjectively. In this paper, we introduce a method that provides an adaptive solution to this problem by using a Bayesian Principle Component Analysis (PCA). This provides an estimate of the data covariance matrix to give a data-driven, non-arbitrary solution to the trade-off between the spatial resolution and the SNR of the beamformer output. This also provides a method for determining when the quality of the data covariance estimate maybe under question. We apply the approach to simulated and real MEG data, and demonstrate the way in which it can automatically adapt the regularization to give good performance over a range of noise and signal levels. PMID- 21620978 TI - Symbol processing in the left angular gyrus: evidence from passive perception of digits. AB - Arabic digits are one of the most ubiquitous symbol sets in the world. While there have been many investigations into the neural processing of the semantic information digits represent (e.g. through numerical comparison tasks), little is known about the neural mechanisms which support the processing of digits as visual symbols. To characterise the component neurocognitive mechanisms which underlie numerical cognition, it is essential to understand the processing of digits as a visual category, independent of numerical magnitude processing. The 'Triple Code Model' (Dehaene, 1992; Dehaene and Cohen, 1995) posits an asemantic visual code for processing Arabic digits in the ventral visual stream, yet there is currently little empirical evidence in support of this code. This outstanding question was addressed in the current functional Magnetic Resonance (fMRI) study by contrasting brain responses during the passive viewing of digits versus letters and novel symbols at short (50 ms) and long (500 ms) presentation times. The results of this study reveal increased activation for familiar symbols (digits and letters) relative to unfamiliar symbols (scrambled digits and letters) at long presentation durations in the left dorsal Angular gyrus (dAG). Furthermore, increased activation for Arabic digits was observed in the left ventral Angular gyrus (vAG) in comparison to letters, scrambled digits and scrambled letters at long presentation durations, but no digit specific activation in any region at short presentation durations. These results suggest an absence of a digit specific 'Visual Number Form Area' (VNFA) in the ventral visual cortex, and provide evidence for the role of the left ventral AG during the processing of digits in the absence of any explicit processing demands. We conclude that Arabic digit processing depends specifically on the left AG rather than a ventral visual stream VNFA. PMID- 21620979 TI - The effects of aging on material-independent and material-dependent neural correlates of contextual binding. AB - Previous behavioral research suggests that older adults exhibit impairments in source memory across a multitude of stimuli and associated details, possibly due to a deficit in contextual binding. However, it is unclear whether this binding deficit results from alterations in processes that are material-independent, processes that are specific to particular materials, or some combination of the two. We used event-related fMRI to investigate the effect of aging on contextual binding and associated neural activity for words and objects. Behavioral data showed similar age-related deficits in source memory accuracy for both words and objects. Imaging results revealed that young and older adults recruited similar networks of regions in support of contextual binding, independent of material, including the lateral prefrontal cortex (PFC) and hippocampus. Material-dependent binding effects were also present for both groups in ventrolateral prefrontal and parahippocampal cortices for words and objects, respectively. Older adults showed smaller material-independent contextual binding effects in frontal regions, namely the left dorsolateral PFC, and larger binding effects in the hippocampus. Further, age-related declines in object processing effects in extrastriate regions correlated with poorer source accuracy for objects. Collectively, these results suggest that age-related source memory deficits may be primarily due to changes in material-independent contextual binding processes. However, these impairments may be further impacted by impoverished item representations in material-specific processing regions. PMID- 21620980 TI - Clinical fMRI: evidence for a 7T benefit over 3T. AB - Despite there being an increasing number of installations of ultra high field MR systems (>3T) in clinical environments, no functional patient investigations have yet examined possible benefits for functional diagnostics. Here we performed presurgical localization of the primary motor hand area on 3T and 7T Siemens scanners with identical investigational procedures and comparable system specific sequence optimizations. Results from 17 patients showed significantly higher functional sensitivity of the 7T system measured via percent signal change, mean t-values, number of suprathreshold voxels and contrast to noise ratio. On the other hand, 7T data suffered from a significant increase of artifacts (ghosting, head motion). We conclude that ultra high field systems provide a clinically relevant increase of functional sensitivity for patient investigations. PMID- 21620981 TI - Catechol-o-methyl transferase (COMT) val158met polymorphism and adolescent cortical development in patients with childhood-onset schizophrenia, their non psychotic siblings, and healthy controls. AB - Non-psychotic individuals at increased risk for schizophrenia show alterations in fronto-striatal dopamine signaling and cortical gray matter maturation reminiscent of those seen in schizophrenia. It remains unclear however if variations in dopamine signaling influence rates of structural cortical maturation in typically developing individuals, and whether such influences are disrupted in patients with schizophrenia and their non-psychotic siblings. We sought to address these issues by relating a functional Val->Met polymorphism within the gene encoding catechol-o-methyltransferase (COMT)-a key enzymatic regulator of cortical dopamine levels-to longitudinal structural neuroimaging measures of cortical gray matter thickness. We included a total of 792 magnetic resonance imaging brain scans, acquired between ages 9 and 22 years from patients with childhood-onset schizophrenia (COS), their non-psychotic full siblings, and matched healthy controls. Whereas greater Val allele dose (which confers enhanced dopamine catabolism and is proposed to aggravate cortical deficits in schizophrenia) accelerated adolescent cortical thinning in both schizophrenia probands and their siblings, it attenuated cortical thinning in healthy controls. This similarity between COS patients and their siblings was accompanied by differences between the two groups in the timing and spatial distribution of disrupted COMT influences on cortical maturation. Consequently, whereas greater Val "dose" conferred persistent dorsolateral prefrontal cortical deficits amongst affected probands by adulthood, cortical thickness differences associated with varying Val dose in non-psychotic siblings resolved over the age-range studied. These findings suggest that cortical abnormalities in pedigrees affected by schizophrenia may be contributed to by a disruption of dopaminergic infleunces on cortical maturation. PMID- 21620982 TI - NOS1 ex1f-VNTR polymorphism influences prefrontal brain oxygenation during a working memory task. AB - Nitric oxide (NO) synthase produces NO, which serves as first and second messenger in neurons, where the protein is encoded by the NOS1 gene. A functional variable number of tandem repeats (VNTR) polymorphism in the promoter region of the alternative first exon 1f of NOS1 is associated with various functions of human behavior, for example increased impulsivity, while another, non-functional variant was linked to decreased verbal working memory and a heightened risk for schizophrenia. We therefore investigated the influence of NOS1 ex 1f-VNTR on working memory function as reflected by both behavioral measures and prefrontal oxygenation. We hypothesized that homozygous short allele carriers exhibit altered brain oxygenation in task-related areas, namely the dorsolateral and ventrolateral prefrontal cortex and the parietal cortex. To this end, 56 healthy subjects were stratified into a homozygous long allele group and a homozygous short allele group comparable for age, sex and intelligence. All subjects completed a letter n-back task (one-, two-, and three-back), while concentration changes of oxygenated (O(2)Hb) hemoglobin in the prefrontal cortex were measured with functional near-infrared spectroscopy (fNIRS). We found load-associated O(2)Hb increases in the prefrontal and parts of the parietal cortex. Significant load-associated oxygenation differences between the two genotype groups could be shown for the dorsolateral prefrontal cortex and the parietal cortex. Specifically, short allele carriers showed a significantly larger increase in oxygenation in all three n-back tasks. This suggests a potential compensatory mechanism, with task-related brain regions being more active in short allele carriers to compensate for reduced NOS1 expression. PMID- 21620983 TI - Computational neuroergonomics. AB - Neuroergonomics merges neuroscience and ergonomics for the study of brain and behavior in natural and naturalistic settings. Together with the rapid development of neuroergonomics concepts, technologies, and related data, there is an urgent need to develop computational models of neuroergonomics that can help integrate and interpret empirical findings and make predictions for scientific research and practical application. This article discusses the relationship between computational neuroscience and computational neuroergonomics, and describes a queuing network based computational neuroergonomic architecture and its applications. These discussions illustrate the mission and challenges of computational neuroergonomics and future research needs. PMID- 21620984 TI - What difference does a year of schooling make? Maturation of brain response and connectivity between 2nd and 3rd grades during arithmetic problem solving. AB - Early elementary schooling in 2nd and 3rd grades (ages 7-9) is an important period for the acquisition and mastery of basic mathematical skills. Yet, we know very little about neurodevelopmental changes that might occur over a year of schooling. Here we examine behavioral and neurodevelopmental changes underlying arithmetic problem solving in a well-matched group of 2nd (n = 45) and 3rd (n = 45) grade children. Although 2nd and 3rd graders did not differ on IQ or grade- and age-normed measures of math, reading and working memory, 3rd graders had higher raw math scores (effect sizes = 1.46-1.49) and were more accurate than 2nd graders in an fMRI task involving verification of simple and complex two-operand addition problems (effect size = 0.43). In both 2nd and 3rd graders, arithmetic complexity was associated with increased responses in right inferior frontal sulcus and anterior insula, regions implicated in domain-general cognitive control, and in left intraparietal sulcus (IPS) and superior parietal lobule (SPL) regions important for numerical and arithmetic processing. Compared to 2nd graders, 3rd graders showed greater activity in dorsal stream parietal areas right SPL, IPS and angular gyrus (AG) as well as ventral visual stream areas bilateral lingual gyrus (LG), right lateral occipital cortex (LOC) and right parahippocampal gyrus (PHG). Significant differences were also observed in the prefrontal cortex (PFC), with 3rd graders showing greater activation in left dorsal lateral PFC (dlPFC) and greater deactivation in the ventral medial PFC (vmPFC). Third graders also showed greater functional connectivity between the left dlPFC and multiple posterior brain areas, with larger differences in dorsal stream parietal areas SPL and AG, compared to ventral stream visual areas LG, LOC and PHG. No such between-grade differences were observed in functional connectivity between the vmPFC and posterior brain regions. These results suggest that even the narrow one-year interval spanning grades 2 and 3 is characterized by significant arithmetic task-related changes in brain response and connectivity, and argue that pooling data across wide age ranges and grades can miss important neurodevelopmental changes. Our findings have important implications for understanding brain mechanisms mediating early maturation of mathematical skills and, more generally, for educational neuroscience. PMID- 21620985 TI - Neuroplastic changes following rehabilitative training correlate with regional electrical field induced with tDCS. AB - Transcranial direct current stimulation (tDCS) has recently emerged as a promising approach to enhance neurorehabilitative outcomes. However, little is known about how the local electrical field generated by tDCS relates to underlying neuroplastic changes and behavior. To address this question, we present a case study analysis of an individual with hemianopia due to stroke and who benefited from a combined visual rehabilitation training and tDCS treatment program. Activation associated with a visual motion perception task (obtained by functional magnetic resonance imaging; fMRI) was used to characterize local changes in brain activity at baseline and after training. Individualized, high resolution electrical field modeling reproducing precise cerebral and lesioned tissue geometry, predicted distortions of current flow in peri-lesional areas and diffuse clusters of peak electric fields. Using changes in fMRI signal as an index of cortical recovery, correlations to the electrical field map were determined. Significant correlations between the electrical field and change in fMRI signal were region specific including cortical areas under the anode electrode and peri-lesional visual areas. These patterns were consistent with effective tDCS facilitated rehabilitation. We describe the methodology used to analyze tDCS mechanisms through combined fMRI and computational modeling with the ultimate goal of developing a rationale for individualized therapy. PMID- 21620986 TI - Definition of osteoarthritis on MRI: results of a Delphi exercise. AB - OBJECTIVE: Despite a growing body of Magnetic Resonance Imaging (MRI) literature in osteoarthritis (OA), there is little uniformity in its diagnostic application. We envisage in the first instance the definition requiring further validation and testing in the research setting before considering implementation/feasibility testing in the clinical setting. The objective of our research was to develop an MRI definition of structural OA. METHODS: We undertook a multistage process consisting of a number of different steps. The intent was to develop testable definitions of OA (knee, hip and/or hand) on MRI. This was an evidence driven approach with results of a systematic review provided to the group prior to a Delphi exercise. Each participant of the steering group was allowed to submit independently up to five propositions related to key aspects in MRI diagnosis of knee OA. The steering group then participated in a Delphi exercise to reach consensus on which propositions we would recommend for a definition of structural OA on MRI. For each round of voting, >=60% votes led to include and <=20% votes led to exclude a proposition. After developing the proposition one of the definitions developed was tested for its validity against radiographic OA in an extant database. RESULTS: For the systematic review we identified 25 studies which met all of our inclusion criteria and contained relevant diagnostic measure and performance data. At the completion of the Delphi voting exercise 11 propositions were accepted for definition of structural OA on MRI. We assessed the diagnostic performance of the tibiofemoral MRI definition against a radiographic reference standard. The diagnostic performance for individual features was: osteophyte C statistic=0.61, for cartilage loss C statistic=0.73, for bone marrow lesions C statistic=0.72 and for meniscus tear in any region C statistic=0.78. The overall composite model for these four features was a C statistic=0.59. We detected good specificity (1) but less optimal sensitivity (0.46) likely due to detection of disease earlier on MRI. CONCLUSION: We have developed MRI definition of knee OA that requires further formal testing with regards their diagnostic performance (especially in datasets of persons with early disease), before they are more widely used. Our current analysis suggests that further testing should focus on comparisons other than the radiograph, that may capture later stage disease and thus nullify the potential for detecting early disease that MRI may afford. The propositions are not to detract from, nor to discourage the use of traditional means of diagnosing OA. PMID- 21620987 TI - Isolation of microbial pathogens of subclinical mastitis from raw sheep's milk of Epirus (Greece) and their role in its hygiene. AB - The natural raw milk microflora is a factor that expresses its sensorial characteristics. The microbial charge into the mammary gland of healthy animal is low and the application of right and healthy conditions during milking and cheese making procedure, prevents from contaminating as well as maintains the natural microflora in order to lend the particular characteristics of milk. The purpose of the present project was the study of the Total Viable Count (T.V.C.) and the count of total psychrotropic bacteria of raw sheep milk from Boutsiko and Karamaniko breeds, collected from healthy animals, as well as the isolation, identification and enumeration of pathogenic bacteria related with the hygiene and the quality of raw sheep milk (with a particular interest in bacteria that may cause human infection). During the experiment we examined two hundred forty (240) samples of raw sheep milk. In these samples a) Staphylococcus aureus, Salmonella sp., Escherichia coli, Clostridium perfringens (vegetative cells and spores) and Bacillus sp. were isolated and identified b) the Total Viable Count and the total number of psychrotropic bacteria were also specified. The sampling, the preparation of samples and decimal dilutions were based on international methods. The Total viable count was determined using the standard methods of the American Public Health Association, 2002. The total number of psychrotropic bacteria was determined using APHA 1976, 1978 rules. The identification of the bacteria was carried out according to the Bergey's manual. Microscopic examination of Gram stained cells, catalase, oxidase and biochemical tests were performed when necessary to further identify. From the 240 milk samples tested, only 5% were E. coli positive, with mean counts ranged from 2 * 10(3) to 2.4 * 10(4) cfu/ml. S. aureus was isolated from 24% of the samples and the mean count per ml was ranged from <10 to 3.4 * 10(2). Meanwhile, Bacillus spp. was also detected in 29% samples. Vegetative forms and spores of C. perfringens were detected in 13% and 63% of the samples respectively. However, microbiological analyses revealed the presence of a small number of selected pathogens in milk samples such as Salmonella, which was only detected in 5% of the samples. Listeria sp., Pseudomonas sp. and Vibrio cholerae were never found. From the experimental results, the Total Viable Count from raw sheep milk samples, fulfils the microbiological criteria of EU Legislation in a percentage of approximately 97%. PMID- 21620988 TI - Allogeneic hematopoietic cell transplantation for chronic myelofibrosis in Australia and New Zealand: older recipients receiving myeloablative conditioning at increased mortality risk. AB - This retrospective registry analysis examined predictive factors for outcome in 57 patients who underwent allogeneic or syngeneic hematopoietic cell transplantation (HCT) for chronic myelofibrosis (CM), either primary (n = 49) or following an antecedent condition (n = 8), reported to the Australasian Bone Marrow Transplant Registry (ABMTRR) between 1993 and 2005. During the 6 years 2000 to 2005, 40 HCTs were performed for CM compared with 17 in the 7 years 1993 to 1999. Twenty-four recipients (42%) were age 50 or over at transplantation; all of these patients were transplanted after 1997, and 15 were given reduced intensity conditioning (RIC) pretransplantation. The cumulative incidence of transplantation-related mortality was 18% at 100 days and 25% at 1 year posttransplantation. Up to 1 year posttransplantation 16 patients died, with the most common causes being infection (n = 6) and graft-versus-host disease (GVHD) (n = 5). A total of 27 patients survived for 3 years or longer posttransplantation. None of these patients required regular red blood cell transfusions, and of the 17 who had not had splenectomies, none had detectable splenomegaly. Twelve patients had no detectable bone marrow fibrosis, 7 had grade 1 fibrosis, and in 8 patients no information was available. The overall survival (OS) probability for all patients was 72% at 1 year and 58% at 5 years posttransplantation. Patients age 50 and over who received myeloablative conditioning fared poorly, with 1-year overall actuarial survival of 44% compared with 77% for all other patients (P = .007). In multivariate analysis, age 50 years and over at transplantation was the only significant independent unfavorable risk factor for survival post-HCT (hazard ratio 2.71, 95% confidence interval 1.16-6.34, P = .02). This study shows a clear increase in annual numbers of allogeneic HCT performed for CM in Australia and New Zealand in recent years. Five-year survival was favorable compared with international studies, but for older recipients who received myeloablative conditioning, mortality risk was elevated. PMID- 21620989 TI - Improved survival after allogeneic hematopoietic stem cell transplantation in recent years. A single-center study. AB - We analyzed the outcome of allogeneic hematopoietic stem cell transplantation (HSCT) over the past 2 decades. Between 1992 and 2009, 953 patients were treated with HSCT, mainly for a hematologic malignancy. They were divided according to 4 different time periods of treatment: 1992 to 1995, 1996 to 2000, 2001 to 2005, and 2006 to 2009. Over the years, many factors have changed considerably regarding patient age, diagnosis, disease stage, type of donor, stem cell source, genomic HLA typing, cell dose, type of conditioning, treatment of infections, use of granulocyte-colony stimulating factor (G-CSF), use of mesenchymal stem cells, use of cytotoxic T cells, and home care. When we compared the last period (2006 2009) with earlier periods, we found slower neutrophil engraftment, a higher incidence of acute graft-versus-host disease (aGVHD) of grades II-IV, and less chronic GVHD (cGHVD). The incidence of relapse was unchanged over the 4 periods (22%-25%). Overall survival (OS) and transplant-related mortality (TRM) improved significantly in the more recent periods, with the best results during the last period (2006-2009) and a 100-day TRM of 5.5%. This improvement was also apparent in a multivariate analysis. When correcting for differences between the 4 groups, the hazard ratio for mortality in the last period was 0.59 (95% confidence interval [CI]: 0.44-0.79; P < .001) and for TRM it was 0.63 (CI: 0.43-0.92; P = .02). This study shows that the combined efforts to improve outcome after HSCT have been very effective. Even though we now treat older patients with more advanced disease and use more alternative HLA nonidentical donors, OS and TRM have improved. The problem of relapse still has to be remedied. Thus, several different developments together have resulted in significantly lower TRM and improved survival after HSCT over the last few years. PMID- 21620990 TI - Variation of a carotenoid-based trait in relation to oxidative stress and endocrine status during the breeding season in the Eurasian kestrel: a multi factorial study. AB - Carotenoid-based skin colorations vary seasonally in many bird species and are thought to be honest sexually selected signals. In order to provide more insight in the potential signal function and underlying mechanisms of such colorations we here quantified patterns of variation of leg coloration in adult male and female Eurasian kestrels (Falco tinnunculus tinnunculus) over the breeding season, and evaluated the relationship between coloration and levels of carotenoids, androgens and estrogens, oxidative damage and plasma non-enzymatic antioxidant capacity. We studied both reproducing wild and non-reproducing captive birds to test for the effect of diet and breeding effort. Males were more colored than females only during mating, and independently of diet, suggesting that leg-color is a sexually selected trait. Seasonal variation in leg color was associated with circulating carotenoids, but concentrations of these molecules were not related to antioxidant capacity, body condition or oxidative damage. These results indicate that carotenoid-based colorations may not be an honest signal of health status in this species. Production of carotenoid rich eggs coincided with low levels of circulating carotenoids in females, indicating that carotenoids might be a limited resource for laying female kestrels. Finally, young rearing males had higher levels of oxidative damage than females, and wild birds of both sexes had higher levels of these parameters than captive birds. These results may indicate that parental effort and physical activity are costly, independently from hormonal status. Since androgens did not explain carotenoid variation we suggest that multiple interacting factors can regulate carotenoid levels along the season. PMID- 21620991 TI - Diverse LEA (late embryogenesis abundant) and LEA-like genes and their responses to hypersaline stress in post-diapause embryonic development of Artemia franciscana. AB - From post-diapause cysts of Artemia franciscana, we defined fourteen LEA (late embryogenesis abundant) and LEA-like genes, including four novel members (Afrlea1 5, Afrlea3-5, Afrlea3-like1 and Afrlea3-like2), which were classified into four groups: G1, G3, G3-like (LEA group3-like), and SMP-like (seed-maturation-protein like), based on their conserved and diversified sequence motifs and amino acid compositions among bacteria, plants, and animals. We also validated six representative genes based on quantitative RT-PCR, including three LEA and two LEA3-like genes that are down-regulated when dehydrated cysts hatch to desiccation-intolerant larvae as well as one SMP-like gene that is slightly up regulated. We further tested their responses to hypersaline stress for four representatives-one from each group-and found that the expression of Afrlea1-5 and Afrlea3-2 were inducible but not Afrlea3-like1 and Afrsmp-like. This result suggested that the LEA and LEA-like genes may play different roles in resistance to hypersaline stress. PMID- 21620992 TI - Involvement of the G3P shuttle and beta-oxidation pathway in the control of TAG synthesis and lipid accumulation in Yarrowia lipolytica. AB - The oleaginous yeast Yarrowia lipolytica can accumulate up to 38% of its dry weight (DW) as lipids. Factors involved in lipid accumulation, particularly triglycerides, are not well identified. Using different mutations in the glycerol 3-phosphate (G3P) shuttle pathway (Deltagut2 affecting the anabolic dehydrogenase or overexpressing GPD1 affecting the catabolic dehydrogenase), we were able to modulate G3P concentration. We show that in a Po1d genetic background, GPD1 overexpression, GUT2 inactivation or both mutations together result in 1.5, 2.9, and 5.6-fold respective increases in the level of G3P leading to an increase of triacylglyceride (TAG) accumulation. Moreover, our results indicate that each strain with an increased concentration of G3P, also presented a decreased concentration of glycerol. Analysis of the different genes involved in glycerol metabolism indicated that Y. lipolytica does not possess a gene for glycerol-3 phosphatase. These findings suggest that Y. lipolytica has a modified and unique metabolism of glycerol that is dedicated to G3P synthesis (and also to TAG synthesis) which may contribute to its oleaginous character. Furthermore, coupling the G3P shuttle disorders to a deficient beta-oxidation pathway (by inactiving POX1-6 or MFE1 genes) increased TAG and free fatty acids content. Finally, we obtained strains that accumulated up to 65-75% of their DW as lipid. Transcriptional analysis in these strains, revealed that the high levels of lipids resulted from over-expression of genes involved in TAG synthesis (SCT1, encoding a sn-1 acyltransferase; and DGA1, encoding an acylCoA diacylglycerol acyltransferase) and the repression of genes involved in the degradation of TAG (TGL3 and TGL4, encoding triacylglycerol lipases). These findings indicate that TAG synthesis is limited by the availability of G3P and fatty acids, and that the expression of genes involved in TAG homeostasis is regulated by the G3P shuttle and the beta-oxidation pathway. Finally, the synergistic contribution of acyltransferase gene expression to G3P synthesis is required for high levels of TAG synthesis and lipid accumulation in Y. lipolytica. PMID- 21620993 TI - Engineering Escherichia coli for efficient production of 5-aminolevulinic acid from glucose. AB - 5-Aminolevulinic acid (ALA) recently received much attention due to its potential applications in many fields. In this study, we developed a metabolic strategy to produce ALA directly from glucose in recombinant Escherichia coli via the C5 pathway. The expression of a mutated hemA gene, encoding a glutamyl-tRNA reductase from Salmonella arizona, significantly improved ALA production from 31.1 to 176mg/L. Glutamate-1-semialdehyde aminotransferase from E. coli was found to have a synergistic effect with HemA(M) from S. arizona on ALA production (2052mg/L). In addition, we identified a threonine/homoserine exporter in E. coli, encoded by rhtA gene, which exported ALA due to its broad substrate specificity. The constructed E. coli DALA produced 4.13g/L ALA in modified minimal medium from glucose without adding any other co-substrate or inhibitor. This strategy offered an attractive potential to metabolic production of ALA in E. coli. PMID- 21620994 TI - Acute retinal arterial occlusive disorders. AB - The initial section deals with basic sciences; among the various topics briefly discussed are the anatomical features of ophthalmic, central retinal and cilioretinal arteries which may play a role in acute retinal arterial ischemic disorders. Crucial information required in the management of central retinal artery occlusion (CRAO) is the length of time the retina can survive following that. An experimental study shows that CRAO for 97min produces no detectable permanent retinal damage but there is a progressive ischemic damage thereafter, and by 4h the retina has suffered irreversible damage. In the clinical section, I discuss at length various controversies on acute retinal arterial ischemic disorders. Classification of acute retinal arterial ischemic disorders: These are of 4 types: CRAO, branch retinal artery occlusion (BRAO), cotton wool spots and amaurosis fugax. Both CRAO and BRAO further comprise multiple clinical entities. Contrary to the universal belief, pathogenetically, clinically and for management, CRAO is not one clinical entity but 4 distinct clinical entities - non-arteritic CRAO, non-arteritic CRAO with cilioretinal artery sparing, arteritic CRAO associated with giant cell arteritis (GCA) and transient non arteritic CRAO. Similarly, BRAO comprises permanent BRAO, transient BRAO and cilioretinal artery occlusion (CLRAO), and the latter further consists of 3 distinct clinical entities - non-arteritic CLRAO alone, non-arteritic CLRAO associated with central retinal vein occlusion and arteritic CLRAO associated with GCA. Understanding these classifications is essential to comprehend fully various aspects of these disorders. Central retinal artery occlusion: The pathogeneses, clinical features and management of the various types of CRAO are discussed in detail. Contrary to the prevalent belief, spontaneous improvement in both visual acuity and visual fields does occur, mainly during the first 7 days. The incidence of spontaneous visual acuity improvement during the first 7 days differs significantly (p<0.001) among the 4 types of CRAO; among them, in eyes with initial visual acuity of counting finger or worse, visual acuity improved, remained stable or deteriorated in non-arteritic CRAO in 22%, 66% and 12% respectively; in non-arteritic CRAO with cilioretinal artery sparing in 67%, 33% and none respectively; and in transient non-arteritic CRAO in 82%, 18% and none respectively. Arteritic CRAO shows no change. Recent studies have shown that administration of local intra-arterial thrombolytic agent not only has no beneficial effect but also can be harmful. Prevalent multiple misconceptions on CRAO are discussed. Branch retinal artery occlusion: Pathogeneses, clinical features and management of various types of BRAO are discussed at length. The natural history of visual acuity outcome shows a final visual acuity of 20/40 or better in 89% of permanent BRAO cases, 100% of transient BRAO and 100% of non arteritic CLRAO alone. Cotton wools spots: These are common, non-specific acute focal retinal ischemic lesions, seen in many retinopathies. Their pathogenesis and clinical features are discussed in detail. Amaurosis fugax: Its pathogenesis, clinical features and management are described. PMID- 21620995 TI - A pilot study on the DNA-protective, cytotoxic, and apoptosis-inducing properties of olive-leaf extracts. AB - Leaves of olive trees are an abundant raw material in the Mediterranean basin. They contain large amounts of potentially useful phytochemicals and could play beneficial roles in health care. In the present study, the principal bioactive phenols in olive-leaf extracts (OLEs) have been identified and quantified, and their genotoxic/antigenotoxic, cytotoxic and apoptotic effects have been assessed. The Somatic Mutation and Recombination Test (SMART) in wing imaginal discs of Drosophila melanogaster has been performed to test the possible genotoxicity of overall OLE and the individual components oleuropein and luteolin at different concentrations. The same assay was able to detect antigenotoxic activity against hydrogen peroxide as oxidative genotoxicant. None of the extracts/phenols tested showed significant mutagenic activity. This fact, together with the antigenotoxic activity against H(2)O(2) detected for all these extracts/phenols, confirmed the safety of OLE, oleuropein and luteolin in terms of DNA protection. HL60 human promyelocytic leukemia cells were used to assess the cytotoxic effects of the extracts/phenols. OLE, oleuropein and luteolin showed a dose-dependent cytotoxic effect with different IC50 (10MUl/ml, 170MUM, and 40MUM, respectively). DNA fragmentation patterns and cell staining with acridine orange and ethidium bromide indicated that the mechanism for the cytotoxic effect of OLE, oleuropein and luteolin was the apoptotic pathway, with DNA laddering and cytoplasmic and nuclear changes. These results could help explain the mechanism of action that underlies the beneficial effect of OLE, proposed as a nutraceutical in the prevention of human cancer. PMID- 21620996 TI - Induction of micronuclei and nuclear abnormalities in erythrocytes of mosquito fish (Gambusia affinis) following exposure to the pyrethroid insecticide lambda cyhalothrin. AB - In the present study the induction of micronuclei (MN) and nuclear abnormalities (NA) in erythrocytes of mosquitofish (Gambusia affinis) (Baird & Girard 1853) was studied. Fish were exposed to three different concentrations of lambda cyhalothrin (LCT) (1*10(-4)MUg/l, 2*10(-4)MUg/l, 4*10(-4)MUg/l) for periods of 6, 12, 24, and 48h. NA (notched, lobed, blebbed nuclei), MN, bi-nucleated cells, and the ratio of polychromatic erythrocytes (PCEs) to normochromatic erythrocytes (NCEs) were evaluated to assess genotoxicity and cytotoxicity. LCT significantly induced MN and NA in erythrocytes of G. affinis. The PCE/NCE ratio was also decreased after 24- and 48-h treatments of 4*10(-4)MUg/l LCT. The results show that LCT has genotoxic and cytotoxic potential on G. affinis. PMID- 21620997 TI - Metallothioneins and energy budget indices in cadmium and copper exposed spiders Agelena labyrinthica in relation to their developmental stage, gender and origin. AB - The aim of our studies was to explain the role of metallothioneins (MTs) in the neutralization of excessive amounts of metals (essential: copper (Cu) and toxic: cadmium (Cd)) and to describe the energy status in metal-exposed spiders Agelena labyrinthica in relation to its developmental stage, gender and origin. Juvenile, female and male spiders were collected from three variously polluted habitats, transferred to the laboratory and exposed to the metals in their diet. Cu and Cd accumulation in the body and exuviae, bioaccumulation factor, percentage of metallothionein positive cells, MT concentration, percentage of cells with depolarized mitochondria, ATP concentration and ADP/ATP ratio were measured and calculated. Cu appeared to be regulated and its excess is eliminated via, among others, the molting process, while Cd was rather accumulated by the spiders. The level of MTs increased significantly mainly in females exposed to both metals, irrespectively of the pollution degree of their site of origin, indicating a defensive role of the proteins. In general, even if both the MT level and the energy status indices were positively correlated with Cd and Cu concentrations in the spider body, the energy status of A. labyrinthica did not seem disturbed. PMID- 21620998 TI - Transmitted drug resistance and phylogenetic analysis of HIV CRF01_AE in Northern Vietnam. AB - The HIV epidemic in Vietnam began in injecting drug users (IDUs), but increasingly affects the general population. It is therefore important to monitor the spread of infection and, since antiretroviral therapy (ART) is now used more frequently, the prevalence of transmitted drug resistance. Sixty-three 1000 bp pol-gene sequences were generated from treatment-naive HIV-1 CRF01_AE infected patients from four clinics in Northern Vietnam. Four drug resistance mutations; Y181C, L210W, L74I and V75M, were found in four different patients, giving a prevalence of 6.3% (4/63). Earlier studies have shown a lower prevalence and the transmission rate should be regularly monitored prospectively in Vietnam. Additional CRF01_AE (N=190) and outgroup subtype B sequences (N=4) were retrieved from databases and included for phylogenetic analysis and calculations of the time of the most recent common ancestor (tMRCA). The 63 samples from our study clustered into two distinct groups; one small clade (N=3) that had a tMRCA in year 1997.5 and a larger group with an estimated tMRCA in 1989.8. The Vietnamese samples in the large group were distinct from CRF01_AE sequences from Thailand, but closely related to previously sequenced isolates from Vietnam, southern China and the Czech Republic, while the samples in the smaller clade appeared to represent a more recent introduction from Southern Vietnam. Our results showed that sequences from IDUs were intermingled with sequences from sexually infected patients, indicating frequent exchange of virus between the transmission risk groups in Northern Vietnam. PMID- 21620999 TI - An increase in B cell apoptosis by interfering BAFF-BAFF-R interaction with small synthetic molecules. AB - B cell-activating factor (BAFF) transmitted signals through binding to specific BAFF receptors (BAFF-R) to regulate B cell survival and development. We used MTT assay to examine the cytotoxicity of chemicals, flow cytometry analysis to measure BAFF-BAFF-R interactions, and western blotting to detect BAFF protein. Here, we established screening method to find specific compounds to interfere with BAFF-BAFF-R interactions in WIL2-NS B lymphoblast cells. According to screening (imidazol-4-ylcarbonyl)guanidine or (oxazol-4-ylcarbonyl)guanidine derivatives, we selected KR32592, KR32673, KR33232, KR33341 and KR33426 as candidates to interfere with BAFF-BAFF-R interaction. No cytotoxicity was detected by KR32592, KR33232, and KR33426 at the concentration of 5 MUM, and by KR32673, and KR33341 at the concentration of 0.5 MUM. Cell population with BAFF BAFF-R interactions was reduced by the pre-incubation of chemicals with human BAFF-murine CD8 (BAFF-muCD8). Cell population with BAFF-BAFF-R interactions was also decreased by pre-exposure of WIL2-NS cells to chemicals prior to the incubation with BAFF-muCD8. Chemicals also inhibited LPS-stimulated BAFF production from splenocytes. All these effects of chemicals may contribute to the inhibition of BAFF-mediated anti-apoptosis. These data demonstrate that chemicals interfering with BAFF-BAFF-R interaction may be screened with our experimental condition. It suggests that BAFF-BAFF-R interaction could be a chemical target to develop therapeutics for BAFF-mediated autoimmune diseases. PMID- 21621000 TI - Human FoxP3+ regulatory T cells in systemic autoimmune diseases. AB - Since the characterization of CD4(+)CD25(+) regulatory T (Treg) cells in mice, significant progress has been made in the definitions of the phenotype and the function of human Treg cells in health and in pathological conditions. Recent advances in the field leading to a better molecular definition of Treg subsets in humans and the description of the dynamics of differentiation of Treg cells should bring new insights in the understanding of human chronic systemic autoimmune diseases. How Treg cells are compromised in these diseases is a challenging issue because the elucidation of the mechanisms leading to such anomaly might lead to promising novel therapeutic approaches. PMID- 21621001 TI - Autoimmunity: when the immune system becomes the self-ish giant. PMID- 21621002 TI - Effects of vitamin D on the peripheral adaptive immune system: a review. AB - Epidemiological studies have shown that a poor vitamin D status is associated with an increased risk of several diseases, including autoimmune diseases. The immune regulatory function of vitamin D is thought to have an important role in these associations. Cells of the adaptive immune system have shown to be direct targets of the vitamin D metabolites. Besides being direct targets, cells of the adaptive immune system express the enzymes involved in the metabolism of vitamin D, enabling them to locally convert 25(OH)D into its active metabolite 1,25(OH)2D. In this review, the effects of vitamin D on cells of the adaptive immune system are described. Experimental data in vitro show that vitamin D skews cells of the adaptive immune system toward a more tolerogenic status which might be exploited in the treatment of autoimmune diseases. However, it should be noticed that in vivo effects may differ from in vitro effects due to the cross talk between different vitamin D sensitive cells, but data support the view that vitamin D is positively involved in maintaining or restoring immune homeostasis. Upcoming vitamin D supplementation trials will further elucidate the in vivo effects of vitamin D on the immune system and its potency to serve as an immune regulating agent in autoimmune diseases. PMID- 21621003 TI - Computational analysis of high-density peptide microarray data with application from systemic sclerosis to multiple sclerosis. AB - Auto-antibodies are implicated in the pathophysiology of various autoimmune diseases. High-density peptide microarrays incubated with human serum can detect antibody reactivities against thousands of peptides. This enables the identification of new auto-antigens and the determination of the parts of protein antigens (epitopes) that are recognized by antibody paratopes. We discuss the utility of peptide microarrays to investigate epitope-antibody-recognitions (EAR) from systemic sclerosis to multiple sclerosis. The technology can help to establish reliable diagnostic and prognostic biomarkers employing a combination of antigenic peptides. We describe the specifics of peptide microarray data and present bioinformatic methods for their analysis. Quality control, data pre processing and the filtering of specific peptides are demonstrated on an example data set. Peptide microarrays representing 24 selected proteins by 3235 overlapping 15mer peptides were used to measure antibodies in serum of 10 patients with limited cutaneous systemic sclerosis (SSC) and 10 healthy blood donors. The data showed a sparse and skewed distribution, and we observed strong individual differences since many peptide sequences were bound by antibodies of only one serum sample. In the sera of the SSc patients, but not of the healthy controls, we found antibodies to three peptides MGPRRRSRKPEAPRR, TPTPGPSRRGPSLGA and GPSRRGPSLGASSHQ that share a similar sequence motif (GP-R/S-RR). These peptides map to two known linear epitopes at the N-terminus of centromere protein A (CENPA), demonstrating the utility of peptide microarrays. Presented experimental and bioinformatic approach can be applied in the same manner for multiple sclerosis research. PMID- 21621004 TI - Immunization in the adult immunocompromised host. AB - The number of patients with impaired immune response has been steadily increasing within the last years, not only with the onset of the AIDS epidemic, but also due to increasing numbers of subjects on immunosuppressive therapies. These patients are at an increased risk for infections, many of which are preventable by immunization. Inactivated vaccines are generally safe in subjects with underlying immunosuppression. However, immune response and protection may be hampered, depending on the extent of immunosuppression. In contrast, live vaccines such as yellow fever, measles, rubella, herpes zoster, and cholera may lead to severe reactions in immunocompromised patients and have been shown to deteriorate some immune-mediated diseases such as multiple sclerosis. Data on the efficacy of vaccines in biological therapies is scarce. Where necessary vaccines should be updated before immunosuppressive therapies are started. To improve the vaccination status several guidelines exist for immunosuppressed patients at risk such as those with rheumatic diseases, asplenia or solid organ and hematopoietic stem cell transplantation. PMID- 21621005 TI - Transverse myelitis. AB - Acute transverse myelitis (ATM) is an etiologically heterogeneous syndrome with acute or subacute onset, in which inflammation of the spinal cord results in neurologic deficits, manifesting as weakness, sensory loss and autonomic dysfunction. It is frequently associated with infectious or systemic autoimmune diseases, but its etiology remains unknown in a substantial portion of cases, which are classified as idiopathic. Unifying diagnostic criteria for idiopathic and disease-associated ATM were proposed in 2002. Although they have been applied to a few cohorts of patients, the limited information provided in the relevant publications has not yet yielded many new insights on the clinical characteristics, disease course, and outcome of adult patients with idiopathic ATM compared to older studies that did not always distinguish between the various etiologies of ATM. There is, however, some new epidemiological data indicating that the incidence of idiopathic ATM is considerably higher, and the female preponderance greater, than previously recognized. In addition, new data on children with ATM show that the prognosis in pediatric patients is not always as benign as previous studies had indicated. The combination of ATM and optic neuritis characterize Devic's syndrome or neuromyelitis optica (NMO). A seminal discovery was the identification of an antibody that is a specific marker not only for NMO, but also of some of its characteristic manifestations in isolation, including longitudinally extensive TM. This has resulted in the proposal that all of the disorders that are associated with NMO-IgG positivity constitute part of an NMO spectrum of disorders. This antibody recognizes aquaporin-4, which represents the most abundant water channel of the central nervous system. There is growing evidence that the antibodies targeting this channel protein have pathogenic potential, thereby providing insights into the possible pathogenetic mechanisms of at least one type of ATM. PMID- 21621006 TI - MicroRNAs in multiple sclerosis and experimental autoimmune encephalomyelitis. AB - MicroRNA (miRNA) are small non-coding RNA molecules about 21-25 nucleotides long. They control gene regulation by translational repression and cleavage. Several studies have shown that many miRNA are associated with the etiology of different diseases. Recent developments in diverse miRNA profiling platforms like microarray and quantitative real-time PCR may enable the identification of specific miRNA as novel diagnostic and predictive markers for various diseases. MiRNAs could even be used as therapeutic drug targets. Multiple sclerosis (MS) is a chronic autoimmune disease affecting the central nervous system. Dysregulated immune system processes result in demyelination of neurons and consequently, electrical impulses that travel along the nerves are disrupted resulting in the impairment of organs. In the past three years, there has been an increased interest in establishing miRNA-based biomarkers for MS. So far, there are six studies on miRNA expression in MS patients in which first miRNAs were discovered as potential disease markers. For instance, one study showed that blood levels of miR-145 can discriminate MS patients from healthy controls, and another showed that active lesions in the brain are characterized by a strong up-regulation of miR-155. Studies on experimental autoimmune encephalomyelitis (EAE), the animal model of MS, further support the significance of miRNA as e.g. mice with miR-155 deletion are highly resistant to EAE. Such investigations help to understand the molecular processes involved in the disease. The identification of miRNA markers that are associated with type of MS, individual disease activity or clinical progression under treatment may open new avenues for early diagnosis and optimized therapy of MS. PMID- 21621007 TI - Autoimmune diseases of the peripheral nervous system. AB - Autoimmune-mediated diseases targeting the peripheral nerve represent a group of disorders often associated with high clinical disability. At present, therapeutic options are limited. The application of innovative and cutting-edge technologies to the study of immune-mediated disorders of the peripheral nervous system (PNS) have generated a better understanding of underlying principles of the organization of the immune network present in the peripheral nerve and its dialogue with the systemic immune system. These insights may foster the development of specific and highly effective therapies for autoimmune diseases of the peripheral nerve. Of great interest in this context is the application of monoclonal antibodies, such as rituximab or alemtuzumab, which in small observational studies provided promising clinical results. But also other immunomodulatory or immunosuppressive drugs used in other indications currently find their way to PNS autoimmunity. Clearly, prospective controlled clinical trials are warranted before making firm conclusions on the feasibility of these innovative therapeutic approaches for treating immune-mediated disease of the peripheral nerve. PMID- 21621008 TI - Autoantibody diagnostics in clinical practice. AB - Disease associated autoantibodies (AAB) are important biomarkers not only to confirm the diagnosis of the respective systemic autoimmune disease but also to diagnose the disease at very early stages (mono- or oligosymptomatic manifestations) or to diagnose the respective disease without the typical clinical manifestations (atypical forms). A confirmation of the diagnosis in early stages is required, if patients should benefit from early therapeutic intervention. Furthermore, AAB determinations are used for prognostic purposes and for monitoring of disease activity or response to therapy. For the advancement of autoantibody diagnostics in clinical practice the following aspects have to be considered: (i) The search for novel clinically relevant AAB and the identification of autoantigenic targets of AAB broadened the spectrum of autoimmune diagnostics and permit the diagnosis of former idiopathic diseases. (ii) To obtain steady diagnostic variables of clinically relevant AAB, the evaluation studies have to be standardized. (iii) Several special features and novel developments of autoantibody diagnostics make correct interpretation of antibody test results increasingly difficult. (iv) Beside standardization of AAB detection methods and quality management efforts the improvement of autoantibody diagnostics depends on further development of diagnostic algorithms including cost-effective multiparametric analyses. PMID- 21621009 TI - Sensory plasticity of the carotid body: role of reactive oxygen species and physiological significance. AB - Recent studies have shown that acute intermittent hypoxia (IH) induces sensory plasticity of the carotid body manifested as sensory long-term facilitation (LTF), which requires prior conditioning with chronic IH and is mediated by reactive oxygen species (ROS). The purpose of this article is to provide a brief review of chronic IH-induced sensory LTF of the carotid body, sources of ROS, mechanisms underlying sensory LTF and its functional significance. Development of sensory LTF requires conditioning with several days of chronic IH. It is completely reversible following re-oxygenation, does not depend on the severity of hypoxia used for IH conditioning, not species specific and is selectively evoked by acute repetitive hypoxia but not by repetitive hypercapnia. Sensory LTF is not associated morphological changes in the carotid body and is expressed in chronic IH treated adult but not in neonatal rat carotid bodies. Chronic IH increases ROS levels in the carotid body involving 5-HT mediated activation of NADPH oxidase-2 (NOX2) and subsequent inhibition of the mitochondrial complex I. Sensory LTF can be prevented by inhibitors of 5-HT receptors, NOX inhibitors as well as by anti-oxidants. The signaling pathways mediating the sensory LTF are summarized in the second figure. It is suggested that sensory LTF contributes to the persistent sympathetic excitation under chronic IH. PMID- 21621010 TI - Sensitivity of bronchopulmonary receptors to cold and heat mediated by transient receptor potential cation channel subtypes in an ex vivo rat lung preparation. AB - Changes in airway temperature can result in respiratory responses such as cough, bronchoconstriction and mucosal secretion after cold exposure and hyperventilation after heat exposure. In the present investigation, we examined the activity of bronchopulmonary receptors in response to activators of thermo sensitive transient receptor potential (TS-TRP) cation channels using an ex vivo rat lung preparation. Receptive fields in small bronchioles were probed with von Frey hair monofilaments, warm (50 degrees C) or cold (8 degrees C) saline or saline containing TS-TRP agonists. Among 233 fibers tested, 159 (68.2%) responded to heat (50 degrees C). A large proportion of heat-responsive receptors (107/145) were also activated by capsaicin. Heat and capsaicin-evoked responses were both blocked by TRPV1 antagonist, capsazepine. Only 15.3% of airway receptors responded to cold, which was associated with sensitivity to TRPM8 agonist menthol but not to TRPA1 agonist cinnamaldehyde (CA). Moreover, cold-evoked responses was unaffected by TRPA1 antagonist HC-03001. Our observations suggest that TRPV1 and TRPM8 are involved in transducing heat and cold in the lower respiratory tract, respectively. PMID- 21621011 TI - Beta-asarone inhibits synaptic inputs to airway preganglionic parasympathetic motoneurons. AB - Therapeutic application of Asarum, a herbal medicine that has been used for centuries, reportedly causes acute respiratory disturbance. The responsible constituents, the sites of action, and the mechanisms involved in this side effect are unclear. We investigated the effects of beta-asarone, a volatile constituent of Asarum, on neurotransmission in the medullary respiratory neuronal network using extracellular recording of the rhythmic hypoglossal activity and voltage clamp recordings of the postsynaptic activity of the airway preganglionic parasympathetic motoneurons (APPMs) in vitro. beta-Asarone caused progressive decreases in the duration and area of the hypoglossal bursts in a concentration dependent manner. The frequency and amplitude of the bursts were initially unaltered or temporarily increased, but were then inhibited progressively after prolonged exposure. As with the inhibition of the hypoglossal bursts, the tonic and the phasic excitatory and inhibitory postsynaptic currents in the APPMs were attenuated. These data suggest that the Asarum-caused acute respiratory disturbance involves beta-asarone-induced inhibition of neurotransmission in the medullary respiratory neuronal network. PMID- 21621012 TI - Oxygenation of the placental-fetal unit in humans. AB - This review focuses on the role of oxygen and the changes in oxygen levels in the different trimesters of pregnancy in the human placenta. In the first trimester, the physiological conversion of the spiral arteries restricts maternal blood flow into the intervillous space creating a low oxygen environment for the trophoblast and the embryo. In the second trimester, progressive conversion of the spiral arteries allows unhindered entrance of maternal blood into the intervillous space. In early pregnancy, pathology of spiral artery conversion may promote premature flow of maternal blood resulting in miscarriage. In more advanced pregnancy, incomplete conversion of spiral arteries impairs maternal blood flow to the placenta, causing chronic hypoxia and growth restriction of the fetus. Chronically reduced maternal supply of oxygen to the placental-fetal unit may be partially balanced by metabolic reprogramming of the placenta. Acute impairment of oxygenation in the perinatal period and its effect on the placental-fetal unit will also be discussed. PMID- 21621013 TI - Design and characteristics of a stable protein scaffold for specific binding based on variable lymphocyte receptor sequences. AB - Variable lymphocyte receptors (VLRs) serve as antigen binding proteins in jawless vertebrates. Their relatively low molecular weight makes VLRs an interesting alternative to antibodies in biotechnological applications. A typical VLR comprises several unique motifs called leucine-rich repeats (LRRs). Using consensus approach we designed a novel VLR protein (called dVLR) containing six LRR repeats based on a sea lamprey receptor sequence. The designed protein was expressed in Escherichia coli in a soluble, native form and showed very favorable biophysical properties. Recombinant dVLR is monomeric in solution and preserves its secondary structure within the pH range 3.0 to 11.0 and tertiary structure between pH 4.0 and 10.0. It undergoes reversible thermal denaturation in a broad pH range (4.0 to 10.0). The maximal denaturation temperature of 73.9 degrees C is observed at pH 6.0, 0.3M NaCl. Chemical denaturation of dVLR at pH 7.5 is a cooperative two-state process with a midpoint at 3.3M GdmCl and a very high free energy change of unfolding in the absence of denaturant equal to 14.1kcal/mol. The biophysical properties of dVLR make it highly suitable for biotechnological applications such as generation of specific ligand-binding molecules. PMID- 21621014 TI - Direct-reversible binding of small molecules to G protein betagamma subunits. AB - Heterotrimeric guanine nucleotide-binding proteins (G proteins) composed of three subunits alpha, beta, gamma mediate activation of multiple intracellular signaling cascades initiated by G protein-coupled receptors (GPCRs). Previously our laboratory identified small molecules that bind to Gbetagamma and interfere with or enhance binding of select effectors with Gbetagamma. To understand the molecular mechanisms of selectivity and assess binding of compounds to Gbetagamma, we used biophysical and biochemical approaches to directly monitor small molecule binding to Gbetagamma. Surface plasmon resonance (SPR) analysis indicated that multiple compounds bound directly to Gbetagamma with affinities in the high nanomolar to low micromolar range but with surprisingly slow on and off rate kinetics. While the k(off) was slow for most of the compounds in physiological buffers, they could be removed from Gbetagamma with mild chaotropic salts or mildly dissociating collision energy in a mass-spectrometer indicating that compound-Gbetagamma interactions were non-covalent. Finally, at concentrations used to observe maximal biological effects the stoichiometry of binding was 1:1. The results from this study show that small molecule modulation of Gbetagamma-effector interactions is by specific direct non-covalent and reversible binding of small molecules to Gbetagamma. This is highly relevant to development of Gbetagamma targeting as a therapeutic approach since reversible, direct binding is a prerequisite for drug development and important for specificity. PMID- 21621015 TI - Coupling of protein and environment fluctuations. AB - We review the concepts of protein dynamics developed over the last 35years and extend applications of the unified model of protein dynamics to heat flow and spatial fluctuations in hydrated myoglobin (Mb) powders. Differential scanning calorimetry (DSC) and incoherent neutron scattering (INS) data on hydration Mb powders are explained by the temperature-dependence of the hydration-shell beta(h) process measured by dielectric relaxation spectroscopy (DRS). The unified model explains the temperature dependence of DSC and INS data as a kinetic effect due to a fixed experimental time window and a broad distribution of hydration shell beta(h) fluctuation rates. We review the slaving of large scale protein motions to the bulk solvent alpha process, and the metastability of Mb molecules in glass forming bulk solvent at low temperatures. This article is part of a Special Issue entitled: "Protein Dynamics: Experimental and Computational Approaches". PMID- 21621016 TI - New antibiotic-eluting mesh used for soft tissue reinforcement. AB - The surgical implantation of prostheses for soft tissue repair may be followed by post-operative mesh-related infection, a significant and dramatic complication, that is treated by mesh removal. A new antibiotic-eluting mesh has been manufactured on pre-existing polypropylene prostheses using an airbrush spraying technology. Among the degradable polymers tested as coating agents and drug reservoirs, poly(epsilon-caprolactone) (PCL), which is deposited after heating, provides a homogeneous, regular and smooth shell around the polypropylene filaments of the mesh without dramatically altering the biomechanical properties of the new modified mesh. An anti-infective drug (e.g. ofloxacin) is incorporated into this polymeric coating giving a limited burst effect followed by sustained drug diffusion for several days. An ofloxacin-eluting mesh has demonstrated excellent antibacterial activity in vitro on Escherichia coli adherence, biofilm formation and inhibitory diameter, even with low drug loads. Although further in vivo investigations are required to draw conclusions on the anti-infective effectiveness of the coated mesh, the airbrush coating of ofloxacin-PCL on existing prostheses is already potentially appealing in an effort to decrease post-operative infection. PMID- 21621017 TI - Mechanical and corrosion properties of newly developed biodegradable Zn-based alloys for bone fixation. AB - In the present work Zn-Mg alloys containing up to 3wt.% Mg were studied as potential biodegradable materials for medical use. The structure, mechanical properties and corrosion behavior of these alloys were investigated and compared with those of pure Mg, AZ91HP and casting Zn-Al-Cu alloys. The structures were examined by light and scanning electron microscopy (SEM), and tensile and hardness testing were used to characterize the mechanical properties of the alloys. The corrosion behavior of the materials in simulated body fluid with pH values of 5, 7 and 10 was determined by immersion tests, potentiodynamic measurements and by monitoring the pH value evolution during corrosion. The surfaces of the corroded alloys were investigated by SEM, energy-dispersive spectrometry and X-ray photoelectron spectroscopy. It was found that a maximum strength and elongation of 150MPa and 2%, respectively, were achieved at Mg contents of approximately 1wt.%. These mechanical properties are discussed in relation to the structural features of the alloys. The corrosion rates of the Zn Mg alloys were determined to be significantly lower than those of Mg and AZ91HP alloys. The former alloys corroded at rates of the order of tens of microns per year, whereas the corrosion rates of the latter were of the order of hundreds of microns per year. Possible zinc doses and toxicity were estimated from the corrosion behavior of the zinc alloys. It was found that these doses are negligible compared with the tolerable biological daily limit of zinc. PMID- 21621018 TI - Hexasomy of the Prader-Willi/Angelman critical region, including the OCA2 gene, in a patient with pigmentary dysplasia: case report. AB - Derivatives of chromosome 15, often referred to as inv dup(15), represent the most common supernumerary marker chromosome (SMC). SMC(15)s can be classified into two major groups according to their length: small SMC(15) and large ones. Depending on the amount of euchromatin, the carriers may either present with a normal phenotype or with a recognizable syndrome. Here we describe a patient with severe mental retardation, epilepsy, dysmorphic features and pigmentary dysplasia. His karyotype was 47,XY,+mar[41]/46,XY[9]. Chromosomal fluorescence in situ hybridization (FISH) showed the SMC to be originating from chromosome 15, dicentric and containing four copies of the Prader-Willi/Angelman Syndrome Critical Region (PWACR), including the OCA2 gene. Molecular studies indicated that it is maternally derived. This report supports the previous observations assuming that severity of phenotype in patients with SMC(15) depends on the dosage of the PWACR and that skin pigmentation is correlated to OCA2 gene copy number. PMID- 21621019 TI - Effects of phospholipids on the antioxidant activity of alpha-tocopherol in the singlet oxygen oxidation of canola oil. AB - This study evaluated the effects of phosphatidylcholine (PC) or phosphatidylethanolamine (PE) on the antioxidative activity of alpha-tocopherol during oxidation of canola oil by singlet oxygen at 10 degrees C for seven hours. Singlet oxygen was produced by chlorophyll b (4 ppm) under 1,700 lux. The oxidation of oil was evaluated by headspace oxygen consumption by gas chromatography and peroxide values (POVs). Concentrations of PC, PE, chlorophyll, and alpha-tocopherol were determined by HPLC. PC and PE protected chlorophyll from degradation, but they accelerated the degradation of alpha-tocopherol under singlet oxygen. Contents of PC and PE did not change for seven hours under singlet oxygen. alpha-Tocopherol significantly lowered POV and headspace oxygen consumption of canola oil under singlet oxygen, and its antioxidant activity was increased by the co-presence of PC and PE. PC and PE increased chemical quenching of singlet oxygen by tocopherol in decreasing the oil oxidation. PMID- 21621020 TI - Comparative proteome analyses of host protein expression in response to Enterovirus 71 and Coxsackievirus A16 infections. AB - Enterovirus 71 (EV71) and Coxsackievirus A16 (CA16) are the main etiological agents of Hand, Foot and Mouth Disease (HFMD), a common disease among children and had caused several outbreaks in the Asia-Pacific region. Although being genetically close to each other, EV71 infection can cause serious and fatal neurological complications like encephalitis, myocarditis, acute flaccid paralysis (AFP) and aseptic meningitis, but not in CA16 infections. In this study, the cellular response of host cells infected with EV71 and CA16 was characterized and compared by 2-dimensional proteome analyses. A total of 16 proteins were identified to be differentially expressed in EV71 and CA16-infected host cells. Desmin and HSP27, both indirectly regulate the contraction of muscle cells, were significantly downregulated as a result of EV71 infection, suggesting a link to acute flaccid paralysis. The ability of EV71 to evade host immune system may be due to the downregulation of MHC-I synthesis proteins like protein disulfide isomerase A3 and calreticulin. Proteins such as nucleophosmin, nuclear ribonucleoprotein C, and eukaryotic translation initiation factor 2 were all downregulated significantly, suggesting the rapid shutting down of host translation machinery by EV71. These findings provide insight into the nature of high virulent EV71 infection as compared to CA16. PMID- 21621021 TI - A proteomics view on the role of drought-induced senescence and oxidative stress defense in enhanced stem reserves remobilization in wheat. AB - Drought is one of the major factors limiting the yield of wheat (Triticum aestivum L.) particularly during grain filling. Under terminal drought condition, remobilization of pre-stored carbohydrates in wheat stem to grain has a major contribution in yield. To determine the molecular mechanism of stem reserve utilization under drought condition, we compared stem proteome patterns of two contrasting wheat landraces (N49 and N14) under a progressive post-anthesis drought stress, during which period N49 peduncle showed remarkably higher stem reserves remobilization efficiency compared to N14. Out of 830 protein spots reproducibly detected and analyzed on two-dimensional electrophoresis gels, 135 spots showed significant changes in at least one landrace. The highest number of differentially expressed proteins was observed in landrace N49 at 20days after anthesis when active remobilization of dry matter was observed, suggesting a possible involvement of these proteins in effective stem reserve remobilization of N49. The identification of 82 of differentially expressed proteins using mass spectrometry revealed a coordinated expression of proteins involved in leaf senescence, oxidative stress defense, signal transduction, metabolisms and photosynthesis which might enable N49 to efficiently remobilized its stem reserves compared to N14. The up-regulation of several senescence-associated proteins and breakdown of photosynthetic proteins in N49 might reflect the fact that N49 increased carbon remobilization from the stem to the grains by enhancing senescence. Furthermore, the up-regulation of several oxidative stress defense proteins in N49 might suggest a more effective protection against oxidative stress during senescence in order to protect stem cells from premature cell death. Our results suggest that wheat plant might response to soil drying by efficiently remobilize assimilates from stem to grain through coordinated gene expression. PMID- 21621022 TI - Contribution of proteomics of Leishmania spp. to the understanding of differentiation, drug resistance mechanisms, vaccine and drug development. AB - Leishmania spp., protozoan parasites with a digenetic life cycle, cause a spectrum of diseases in humans. Recently several Leishmania spp. have been sequenced which significantly boosted the number and quality of proteomic studies conducted. Here a historic review will summarize work of the pre-genomic era and then focus on studies after genome information became available. Firstly works comparing the different life cycle stages, in order to identify stage specific proteins, will be discussed. Identifying post-translational modifications by proteomics especially phosphorylation events will be discussed. Further the contribution of proteomics to the understanding of the molecular mechanism of drug resistance and the investigation of immunogenic proteins for the identification of vaccine candidates will be summarized. Approaches of how potentially secreted proteins were identified are discussed. So far 30-35% of the total predicted proteome of Leishmania spp. have been identified. This comprises mainly the abundant proteins, therefore the last section will look into technological approaches on how this coverage may be increased and what the gel free and gel-based proteomics have to offer will be compared. PMID- 21621023 TI - From in silico target prediction to multi-target drug design: current databases, methods and applications. AB - Given the tremendous growth of bioactivity databases, the use of computational tools to predict protein targets of small molecules has been gaining importance in recent years. Applications span a wide range, from the 'designed polypharmacology' of compounds to mode-of-action analysis. In this review, we firstly survey databases that can be used for ligand-based target prediction and which have grown tremendously in size in the past. We furthermore outline methods for target prediction that exist, both based on the knowledge of bioactivities from the ligand side and methods that can be applied in situations when a protein structure is known. Applications of successful in silico target identification attempts are discussed in detail, which were based partly or in whole on computational target predictions in the first instance. This includes the authors' own experience using target prediction tools, in this case considering phenotypic antibacterial screens and the analysis of high-throughput screening data. Finally, we will conclude with the prospective application of databases to not only predict, retrospectively, the protein targets of a small molecule, but also how to design ligands with desired polypharmacology in a prospective manner. PMID- 21621024 TI - Molecular profiling of the human nasal epithelium: A proteomics approach. AB - A comprehensive proteomic profiling of nasal epithelium (NE) is described. This study relies on simple subcellular fractionation used to obtain soluble- and membrane-enriched fractions followed by 2-dimensional liquid chromatography (2D LC) separation and tandem mass spectrometry (MS/MS). The cells were collected using a brushing technique applied on NE of clinically evaluated volunteers. Subsequently, the soluble- and the membrane-protein enriched fractions were prepared and analyzed in parallel using 2D-LC-MS/MS. In a set of 1482 identified proteins, 947 (63.9%) proteins were found to be associated to membrane fraction. Grand average hydropathy value index (GRAVY) analysis, the transmembrane protein mapping and annotations of primary location deposited in the Human Protein Reference Database (HPRD) confirmed an enrichment of hydrophobic proteins on this dataset. Ingenuity Pathway Analysis (IPA) of soluble fraction revealed an enrichment of molecular and cellular functions associated with cell death, protein folding and drug metabolism while in membrane fraction showed an enrichment of functions associated with molecular transport, protein trafficking and cell-to-cell signaling and interaction. The IPA showed similar enrichment of functions associated with cellular growth and proliferation in both soluble and membrane subproteomes. This finding was in agreement with protein content analysis using exponentially modified protein abundance index (emPAI). A comparison of our data with previously published studies focusing on respiratory tract epithelium revealed similarities related to identification of proteins associated with physical barrier function and immunological defence. In summary, we extended the NE molecular profile by identifying and characterizing proteins associated to pivotal functions of a respiratory epithelium, including the control of fluid volume and ionic composition at the airways' surface, physical barrier maintenance, detoxification and immunological defence. The extent of similarities supports the applicability of a less invasive analysis of NE to assess prognosis and treatment response of lung diseases such as asthma, cystic fibrosis and chronic obstructive pulmonary disease. PMID- 21621025 TI - Distinct cell surface proteome profiling by biotin labeling and glycoprotein capturing. AB - We performed here MS-based cell surface proteome profiling of HCT-116 cells by two distinct methods based on biotin labeling and glycoprotein capturing. In total, 742 biotinylated and 219 glycosylated proteins were identified by the biotin labeling and glycoprotein capturing, of which 224 and 138 proteins known to be located on plasma membrane were included, respectively, according to ingenuity pathway analysis. Although 104 plasma membrane proteins were identified by both methods, the rest of 154 were identified only by one. Almost all the identified plasma membrane proteins possessed consensus N-glycosylation sites, and proteins having various numbers of glycosylation sites were identified by both methods. Thus, the discrepancies of the identified proteins obtained from those two methods might not be only due to the number of glycosylation sites, but also to the expression and/or glycosylation level of the cell surface proteins. We also identified 312 N-glycosylated proteins from xenograft samples by glycoprotein capturing of which 135 were known as plasma membrane proteins. Although a number of highly-expressed plasma membrane proteins were common between culture and xenograft cells, some proteins showed culture- or xenograft specific expression, suggesting that those proteins might contribute to grow in different environment. PMID- 21621026 TI - Adenovirus and miRNAs. AB - Adenovirus infection has a tremendous impact on the cellular silencing machinery. Adenoviruses express high amounts of non-coding virus associated (VA) RNAs able to saturate key factors of the RNA interference (RNAi) processing pathway, such as Exportin 5 and Dicer. Furthermore, a proportion of VA RNAs is cleaved by Dicer into viral microRNAs (mivaRNAs) that can saturate Argonaute, an essential protein for miRNA function. Thus, processing and function of cellular miRNAs is blocked in adenoviral-infected cells. However, viral miRNAs actively target the expression of cellular genes involved in relevant functions such as cell proliferation, DNA repair or RNA regulation. Interestingly, the cellular silencing machinery is active at early times post-infection and can be used to control the adenovirus cell cycle. This is relevant for therapeutic purposes against adenoviral infections or when recombinant adenoviruses are used as vectors for gene therapy. Manipulation of the viral genome allows the use of adenoviral vectors to express therapeutic miRNAs or to be silenced by the RNAi machinery leading to safer vectors with a specific tropism. This article is part of a "Special Issue entitled:MicroRNAs in viral gene regulation". PMID- 21621028 TI - An ultrasensitive method for the detection of gene fragment from transgenics using label-free gold nanoparticle probe and dynamic light scattering. AB - The detection of transgenic products is of great significance for the development of transgenic technique. In this paper, we developed a simple, rapid and ultrasensitive method for the detection of sequence-specific Nopaline synthase (NOS) gene from the transgenic plants using label-free gold nanoparticle (NP) probe and dynamic light scattering (DLS) technology. Gold NPs were stable in NaCl solution with the presence of NOS gene probe. On the contrary, they were aggregated in NaCl solution when the probe sequence was hybridized with target sequence. The change in the size of gold NPs can be detected by DLS technology with high sensitivity. Under the optimal conditions, the average hydrodynamic diameter of gold NPs was linear with the concentration of the target sequence ranging from 1.0*10(-13)mol L(-1) to 5.0*10(-9)mol L(-1), with a detection limit of 3.0*10(-14)mol L(-1) (S/N=3). The relative standard deviation (at 1.0*10( 9)mol L(-1) of target sequence) was 4.8% (n=11). The result shows that gold NPs based DLS method has great potential in the analysis of transgenic products. PMID- 21621027 TI - Spatiotemporal delivery of bone morphogenetic protein enhances functional repair of segmental bone defects. AB - Osteogenic growth factors that promote endogenous repair mechanisms hold considerable potential for repairing challenging bone defects. The local delivery of one such growth factor, bone morphogenetic protein (BMP), has been successfully translated to clinical practice for spinal fusion and bone fractures. However, improvements are needed in the spatial and temporal control of BMP delivery to avoid the currently used supraphysiologic doses and the concomitant adverse effects. We have recently introduced a hybrid protein delivery system comprised of two parts: a perforated nanofibrous mesh that spatially confines the defect region and a functionalized alginate hydrogel that provides temporal growth factor release kinetics. Using this unique spatiotemporal delivery system, we previously demonstrated BMP-mediated functional restoration of challenging 8mm femoral defects in a rat model. In this study, we compared the efficacy of the hybrid system in repairing segmental bone defects to that of the current clinical standard, collagen sponge, at the same dose of recombinant human BMP-2. In addition, we investigated the specific role of the nanofibrous mesh tube on bone regeneration. Our results indicate that the hybrid delivery system significantly increased bone regeneration and improved biomechanical function compared to collagen sponge delivery. Furthermore, we observed that presence of the nanofiber mesh tube was essential to promote maximal mineralized matrix synthesis, prevent extra-anatomical mineralization, and guide an integrated pattern of bone formation. Together, these results suggest that spatiotemporal strategies for osteogenic protein delivery may enhance clinical outcomes by improving localized protein retention. PMID- 21621029 TI - A review of separation methods for the determination of estrogens and plastics derived estrogen mimics from aqueous systems. AB - Recent methods of separation and detection for the quantification of trace-level concentrations of selected endocrine disrupting compounds (EDCs) from aqueous systems are reviewed. A brief introduction of the selected EDCs (natural and synthetic estrogens and plastics-derived xenoestrogens), including their characteristics and importance, is presented. Sample preparation and extraction trends are discussed. Various types of separation techniques are presented, with the express goal of emphasizing time and cost-effective methods that isolate and quantify trace-levels of multiple endocrine disruptors from aqueous systems. PMID- 21621030 TI - Recent developments in handheld and portable optosensing-a review. AB - Recent developments in portable and handheld opto-chemical analytical instrumentation over the last decade (2000-2010) are reviewed. First, the characteristics of typical portable/handheld instrumentation are discussed from different points of view: in situ operation, low energy consumption, ease of use, and self-contained devices. These advancements have improved or hastened improvements in the development of miniaturized optoelectronic and optical components, mainly solid-state devices such as different types of semiconductor lasers, light-emitting diodes, and photodiodes. A brief review of advances in these components is also presented. The numerous examples of portable instrumentation presented have been classified according to direct-recognition and reagent-based sensing, and within these, by absorption and emission-based systems. The conclusion discusses some key trends and future perspectives for this technology. PMID- 21621031 TI - Fast determination of thyroid stimulating hormone in human blood serum without chemical preprocessing by using infrared spectroscopy and least squares support vector machines. AB - The least squares support vector machines (LS-SVM) was used to model infrared spectral data for TSH hormone secreted by thyroid, which regulates the basal metabolic rate. This model was used for direct estimation of the content of TSH in blood serum samples, and the results were comparable with those obtained with the conventional analytical method based on chemoluminescence methodology. Excellent agreement was observed between the conventional method and the newly developed calibration model based in analysis of spectral data with LS-SVM. The latter has clear advantages, because it is fast and requires no reagent once the measurements were done directly in the serum by using a simple mid-infrared spectrometer in the ATR mode. An important advantage observed in this calibration method based on LS-SVM is the remarkable capacity to avoid overfitting in the model-building step, that is, the developed method is highly robust. PMID- 21621032 TI - Fast batch injection analysis of H(2)O(2) using an array of Pt-modified gold microelectrodes obtained from split electronic chips. AB - A fast and robust analytical method for amperometric determination of hydrogen peroxide (H(2)O(2)) based on batch injection analysis (BIA) on an array of gold microelectrodes modified with platinum is proposed. The gold microelectrode array (n=14) was obtained from electronic chips developed for surface mounted device technology (SMD), whose size offers advantages to adapt them in batch cells. The effect of the dispensing rate, volume injected, distance between the platinum microelectrodes and the pipette tip, as well as the volume of solution in the cell on the analytical response were evaluated. The method allows the H(2)O(2) amperometric determination in the concentration range from 0.8 MUmolL(-1) to 100 MUmolL(-1). The analytical frequency can attain 300 determinations per hour and the detection limit was estimated in 0.34 MUmolL(-1) (3sigma). The anodic current peaks obtained after a series of 23 successive injections of 50 MUL of 25 MUmolL( 1) H(2)O(2) showed an RSD<0.9%. To ensure the good selectivity to detect H(2)O(2), its determination was performed in a differential mode, with selective destruction of the H(2)O(2) with catalase in 10 mmolL(-1) phosphate buffer solution. Practical application of the analytical procedure involved H(2)O(2) determination in rainwater of Sao Paulo City. A comparison of the results obtained by the proposed amperometric method with another one which combines flow injection analysis (FIA) with spectrophotometric detection showed good agreement. PMID- 21621033 TI - A new method for the determination of short-chain fatty acids from the aliphatic series in wines by headspace solid-phase microextraction-gas chromatography-ion trap mass spectrometry. AB - A new analytical method for the determination of nine short-chain fatty acids (acetic, propionic, isobutyric, butyric, isovaleric, 2-methylbutyric, hexanoic, octanoic and decanoic acids) in wines using the automated HS/SPME-GC-ITMS technique was developed and optimised. Five different SPME fibers were tested and the influence of different factors such as temperature and time of extraction, temperature and time of desorption, pH, strength ionic, tannins, anthocyans, SO(2), sugar and ethanol content were studied and optimised using model solutions. Some analytes showed matrix effect so a study of recoveries was performed. The proposed HS/SPME-GC-ITMS method, that covers the concentration range of the different analytes in wines, showed wide linear ranges, values of repeatability and reproducibility lower than 4.0% of RSD and detection limits between 3 and 257 MUgL(-1), lower than the olfactory thresholds. The optimised method is a suitable technique for the quantitative analysis of short-chain fatty acids from the aliphatic series in real samples of white, rose and red wines. PMID- 21621034 TI - Determination of amphetamine-type stimulants in oral fluid by solid-phase microextraction and gas chromatography-mass spectrometry. AB - A method for the simultaneous identification and quantification of amphetamine (AMP), methamphetamine (MET), fenproporex (FEN), diethylpropion (DIE) and methylphenidate (MPH) in oral fluid collected with QuantisalTM device has been developed and validated. Thereunto, in-matrix propylchloroformate derivatization followed by direct immersion solid-phase microextraction and gas chromatography mass spectrometry were employed. Deuterium labeled AMP was used as internal standard for all the stimulants and analysis was performed using the selected ion monitoring mode. The detector response was linear for the studied drugs in the concentration range of 2-256 ng mL(-1) (neat oral fluid), except for FEN, whereas the linear range was 4-256 ng mL(-1). The detection limits were 0.5 ng mL(-1) (MET), 1 ng mL(-1) (MPH) and 2 ng mL(-1) (DIE, AMP, FEN), respectively. Accuracy of quality control samples remained within 98.2-111.9% of the target concentrations, while precision has not exceeded 15% of the relative standard deviation. Recoveries with QuantisalTM device ranged from 77.2% to 112.1%. Also, the goodness-of-fit concerning the ordinary least squares model in the statistical inference of data has been tested through residual plotting and ANOVA. The validated method can be easily automated and then used for screening and confirmation of amphetamine-type stimulants in drivers' oral fluid. PMID- 21621035 TI - Feasibility of capillary liquid chromatography-microchip-atmospheric pressure photoionization-mass spectrometry for pesticide analysis in tomato. AB - A new and sensitive analytical method, using capillary liquid chromatography (capLC) with a microfabricated heated nebulizer chip for atmospheric pressure photoionization and tandem mass spectrometry (MUAPPI-MS/MS), was developed for the analysis of selected carbamate pesticides in a tomato matrix. The performance of the instrumental method was evaluated, using seven pesticides, namely oxamyl, methomyl, aldicarb, carbofuran, pirimicarb, thiocarb, and ditalimfos. The limits of detection achieved with the capLC-MUAPPI-MS/MS method in the positive ion mode were low, ranging from 0.25 ng mL(-1) for pirimicarb to 5 ng mL(-1) for oxamyl and methomyl, corresponding to 5 and 0.25 MUg kg(-1) for tomato samples, respectively, which are clearly below the maximum residue limits for them in fruits and vegetables. The repeatability of the instrumental method ranged from 2.9 to 13.9% (RSD) at a low (0.05 MUg mL(-1)) concentration level. An adequate linearity (r(2)=0.984-0.999) at a concentration range from 0.005 to 5.0 MUg mL( 1) was observed for all pesticides. The results obtained show that the capLC MUAPPI-MS/MS method developed could be used for the analysis of selected pesticides from tomato. PMID- 21621036 TI - Comparison of near infrared and microwave resonance sensors for at-line moisture determination in powders and tablets. AB - In this paper we demonstrate the feasibility of replacing KF for water content testing in bulk powders and tablets with at-line near infrared (NIR) or microwave resonance (MR) methods. Accurate NIR and MR prediction models were developed with a minimalistic approach to calibration. The NIR method can accurately predict water content in bulk powders in the range of 0.5-5% w/w. Results from this method were compared to a MR method. We demonstrated excellent agreement of both NIR and MR methods for powders vs. the reference KF method. These methods are applicable to in-process control or quality control environments. One of the aims of this study was to determine if a calibration developed for a particular product could be used to predict the water content of another product (with related composition) but containing a different active pharmaceutical ingredient (API). We demonstrated that, contrary to the NIR method, a general MR method can be used to predict water content in two different types of blends. Finally, we demonstrated that a MR method can be developed for at-line moisture determination in tablets. PMID- 21621037 TI - Functionalization of mesoporous silica membrane with a Schiff base fluorophore for Cu(II) ion sensing. AB - A Schiff base (SB) immobilized hybrid mesoporous silica membrane (SB-HMM) was prepared by immobilizing a Schiff base onto the pore surface of mesoporous silica (pore size=3.1 nm) embedded in the pores of a porous anodic alumina membrane. In contrast to the non-fluorescent analogous SB molecule in homogeneous solutions, SB-HMM exhibited intense fluorescence due to emission enhancement caused by aggregation of SB groups on the pore surface. The high quantum efficiency of the surface SB groups allows SB-HMM to function as a fluorescent sensor for Cu(II) ions in an aqueous solution with good sensitivity, selectivity and reproducibility. Under the optimal conditions described, the linear ranges of fluorescence intensity for Cu(II) are 1.2-13.8(M (R(2)=0.993) and 19.4-60 (R(2)=0.992) (M. The limit of detection for Cu(II) is 0.8 MUM on basis of the definition by IUPAC (C(LOD)=3.3S(b)/m). PMID- 21621038 TI - Patterning pallet arrays for cell selection based on high-resolution measurements of fluorescent biosensors. AB - Pallet arrays enable cells to be separated while they remain adherent to a surface and provide a much greater range of cell selection criteria relative to that of current technologies. However there remains a need to further broaden cell selection criteria to include dynamic intracellular signaling events. To demonstrate the feasibility of measuring cellular protein behavior on the arrays using high resolution microscopy, the surfaces of individual pallets were modified to minimize the impact of scattered light at the pallet edges. The surfaces of the three-dimensional pallets on an array were patterned with a coating such as fibronectin using a customized stamping tool. Micropatterns of varying shape and size were printed in designated regions on the pallets in single or multiple steps to demonstrate the reliability and precision of patterning molecules on the pallet surface. Use of a fibronectin matrix stamped at the center of each pallet permitted the localization of H1299 and mouse embryonic fibroblast (MEF) cells to the pallet centers and away from the edges. Compared to pallet arrays with fibronectin coating the entire top surface, arrays with a central fibronectin pattern increased the percentage of cells localized to the pallet center by 3-4-fold. Localization of cells to the pallet center also enabled the physical separation of cells from optical artifacts created by the rough pallet side walls. To demonstrate the measurement of dynamic intracellular signaling on the arrays, fluorescence measurements of high spatial resolution were performed using a RhoA GTPase biosensor. This biosensor utilized fluorescence resonance energy transfer (FRET) between cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP) to measure localized RhoA activity in cellular ruffles at the cell periphery. These results demonstrated the ability to perform spatially resolved measurements of fluorescence-based sensors on the pallet arrays. Thus, the patterned pallet arrays should enable novel cell separations in which cell selection is based on complex cellular signaling properties. PMID- 21621039 TI - Direct classification of olive oils by using two types of ion mobility spectrometers. AB - In this work, we explored the use of an Ion Mobility Spectrometry (IMS) device with an ultraviolet (UV) source, and of a Gas Chromatographic (GC) column coupled to an IM Spectrometer with a tritium source, for the discrimination of three grades of olive oil, namely: extra virgin olive oil (EVOO), olive oil (OO) and pomace olive oil (POO). The three types of oil were analyzed with both equipment combinations as coupled to a headspace system and the obtained ion mobility data were consecutively processed with various chemometric tools. The classification rate for an independent validation set was 86.1% (confidence interval at 95% [83.4%, 88.5%]) with an UV-IMS and 100% (confidence interval at 95% [87%, 100%]) using a GC-IMS system. The classification rate was improved by using a more suitable ionization source and a pre-separation step prior to the IM analysis. PMID- 21621040 TI - A stepwise strategy employing automated screening and DryLab modeling for the development of robust methods for challenging high performance liquid chromatography separations: a case study. AB - A stepwise method development strategy has been employed to develop a robust HPLC method to resolve several closely eluting structurally related impurities in an active pharmaceutical ingredient (API). This strategy consisted of automated column screening, optimization of the most critical chromatographic parameters, DryLab((r)) modeling, and experimental verification of optimized separation conditions. DryLab((r)) was used to predict an optimized gradient profile and separation temperature and these predictions were verified experimentally. A discussion of the accuracy of these predictions is presented. The robustness of the method was verified and the ability of DryLab((r)) to predict, with reasonable accuracy, the outcome of such robustness studies was also examined. Once the robustness was established by the DryLab((r)) predictions the remainder of the subsequent verification by experiment becomes a simple reiterative exercise. This study also demonstrates that factors such as column chemistry and critical chromatographic parameters can have a profound and oftentimes interrelated effect on the chromatographic separation of isomers, bromo analogs and other structurally very similar impurities. Therefore, it is critical to adopt a rational strategy, as demonstrated here, to evaluate the interplay of these factors, thereby greatly enhancing method development efficiency. PMID- 21621042 TI - Allergic disorders interface with ear, nose, and throat disorders. PMID- 21621043 TI - The evolution of understanding inhalant allergy. AB - This article summarizes the developments that led to the current approach to immunotherapy. These developments were characterized in the early years by empirically derived successive approximations to arrive at effective injection regimens, in the middle years by a sorting through of the wide variations in practice with placebo-controlled clinical trials, and more recently by a closer association of clinical and laboratory measures to better define evidence-based practices. The pace of investigation along with the scientific quality continues to increase. PMID- 21621044 TI - Epidemiology of allergy. AB - Atopic disease represents a spectrum of disorders characterized by abnormal sensitivity mediated by IgE; approximately 20% of Americans suffer from some form of allergic disease. The sequelae of inhalant and food allergies may present in many organ systems. Manifestations of allergic disease in one site are often associated with symptoms from another site. It is important for clinicians to understand the epidemiology of atopic disease and its causes to facilitate implementation of effective treatment and prevention strategies. This review focuses on the epidemiology of inhalant allergies causing allergic rhinitis and asthma and on IgE-mediated food allergies. PMID- 21621045 TI - Types of rhinitis. AB - Rhinitis is a familiar disorder affecting up to 20% of the general population. Chronic rhinitis can be broadly classified into allergic, infectious, or nonallergic or noninfectious, which are distinguished mainly by a careful history and allergy testing. The pathophysiology of nonallergic rhinitis likely involves a combination of inflammatory and neurogenic mechanisms that are poorly understood. Treatment involves avoiding the offending agent, when possible, and using appropriate medications to control the patient's predominant symptoms. PMID- 21621046 TI - Differential diagnosis in allergy. AB - The otolaryngic allergist must be able to distinguish between common nonallergic diagnoses that present very similarly to allergic conditions. This article describes a few of the vast myriad of conditions that must be ruled out before a diagnosis of allergy may be made. After reading this article clinicians will be able to identify various conditions, which will enhance their ability to appropriately make correct decisions for prompt and efficient management of their patients with allergic or nonallergic diseases of the head and neck. PMID- 21621047 TI - Immunology of allergy. AB - Knowledge of the immune system is advancing rapidly. This review provides an update on the allergy players-the cells and major mediators-and the form and function of each; discusses how these cells and mediators weave together in the elegant but destructive dance of allergy; and details how specific immunotherapy can cure allergy. PMID- 21621048 TI - Physical findings in allergy. AB - Allergies are typically diagnosed based on detailed history elicited from a patient. Confirmation of the diagnosis by allergy skin or in vitro testing is sometimes also helpful. The authors discuss several physical examination features, specifically in the head and neck region, that are often suggestive of allergy presence. PMID- 21621049 TI - Diagnosis of inhalant allergies: patient history and testing. AB - In the United States, roughly 20% to 25% of the general adult population is afflicted by some form of chronic allergic respiratory disease, making allergy one of the most commonly diagnosed disorders. Among children, allergic disease is more common, with some sources estimating that it affects up to 40% of children. The focus of this article involves making the diagnosis of the most familiar and best understood of the hypersensitivity reactions, type 1 hypersensitivity, also termed immediate hypersensitivity. Although type 1 hypersensitivity can be caused by ingestion of food antigens or pharmaceuticals, this article focuses on IgE mediated allergic disease caused primarily by inhalant allergens. PMID- 21621050 TI - Role of allergy in sleep-disordered breathing. AB - Sleep-related symptoms are extremely common in patients with allergic rhinitis. Sleep impairment is likely a major contributor to the overall disease morbidity, direct and indirect health care costs, and the loss of work productivity associated with allergic rhinitis. The association between allergic rhinitis and sleep, and the subsequent impact on disease-specific and general health quality of life measures, is well documented in large epidemiologic studies as well as controlled clinical trials. This article focuses on sleep disruption caused by allergic rhinitis, and the therapeutic and surgical options available to tackle the problem. PMID- 21621051 TI - The role of allergy in otitis media with effusion. AB - The role of allergy in chronic otitis media with effusion (OME) is controversial. New evidence from cellular biology and immunology explain the basics of allergic reactions and allow more accurate diagnosis of allergies and inflammatory disease throughout the unified airway. This article examines the epidemiologic, methodological, and immunologic studies of allergic causes of OME, including (1) evidence for and against OME as an allergic disease, (2) allergy as a cause for eustachian tube obstruction, (3) examination of the most sensitive diagnostic tests for allergy, and (4) the effect of treatment of underlying allergies in improving and resolving middle ear disease. PMID- 21621052 TI - Allergic and immunologic features of Meniere's disease. AB - Meniere's disease (MD), which by definition is idiopathic, has been ascribed to various causes, including both inhalant and food allergies. Patients with MD report higher rates of allergy history and positive skin or in vitro tests compared with a control group of patients with other otologic diseases and to the general public. Recent immunologic studies have shown higher rates of circulating immune complexes, CD4, and other immunologic components in patients with MD compared with healthy controls. Published treatment results have shown benefit from immunotherapy and/or dietary restriction for symptoms of MD in those patients who present with both allergy and MD. PMID- 21621053 TI - Asthma. AB - This article presents the complexity of asthma and its ensuing management. The author details the clinical presentation of asthma, discussing its variability of presentation over time and within and among individuals. Pathophysiology is discussed, including immunohistopathologic features and inflammatory mediators, risk factors, comorbid conditions, natural history, and triggers. Diagnosis is presented in light of the challenging history of patients suspected of the disorder; pulmonary function testing, symptom assessment, impairment assessment, and risk assessment. Categories of asthma severity, controlling triggers, pharmacotherapy, stepwise approaches, and ongoing assessment are discussed, with a review of a clinical trial assessing efficacy of management. PMID- 21621054 TI - Nasal polyps: pathogenesis and treatment implications. AB - Nasal polyps (NPs) represent a common clinical end point for a myriad of inflammatory disease processes involving the paranasal sinuses. Chronic rhinosinusitis is the most common cause for NPs, but not all NPs are created equally. This article outlines the current understanding of pathogenesis in nasal polyposis and discusses the implications on therapy. PMID- 21621055 TI - Allergic fungal rhinosinusitis. AB - Allergic fungal rhinosinusitis is a phenotype of chronic rhinosinusitis with nasal polyposis, characterized by type 1 hypersensitivity to fungi, eosinophilic mucin with fungal hyphae in sinus secretions, and propensity for mucocele formation and bone erosion. Although its differentiation from other forms of chronic polypoid rhinosinusitis with eosinophilic mucin is sometimes problematic, type 1 hypersensitivity is a component of the disease process. Medical and surgical management can be augmented by immunotherapy directed toward the patient's specific allergen sensitivities. The primary rationale for immunotherapy is to control the allergic diathesis that may be contributing to the patient's chronic sinus inflammation. PMID- 21621056 TI - Allergy treatment: environmental control strategies. AB - This article aims to help physicians and allergy care providers understand: the role of environmental control in the treatment of allergic disease; the concept of "the inflammatory load"; current published studies on environmental control; factors that influence levels of indoor and outdoor allergens; different methods to decrease patients' exposure to indoor and outdoor allergens; problems related to nonallergic symptom triggers; special considerations for school and workplace avoidance; role of environmental control in the prevention of allergic disease; various products available on the market to assist in avoidance; and how to plan with the patient to implement environmental control strategies. PMID- 21621057 TI - Pharmacotherapy for allergic rhinitis. AB - Allergic rhinitis affects millions of Americans and the numbers continue to increase. Fortunately, there exists a wide array of pharmacotherapeutic options with relatively safe side effect profiles for the management of the varying subtypes. Additionally, there are newer agents on the horizon. The efficacies of intranasal corticosteroids, antihistamines, combination topical therapy, leukotriene inhibitors, mast cell stabilizers, anticholinergics, mucolytics, decongestants, and anti-IgE are reviewed. PMID- 21621058 TI - Immunotherapy--traditional. AB - Immunotherapy is an excellent treatment option for a selected subset of patients with inhalant allergies. It consists of intentional serial exposures to allergens, which modulate the immune system and induce immune tolerance through down-regulating the allergic response, resulting in an overall decrease in symptoms. Immunotherapy has been shown to have long-term efficacy in the management of inhalant allergies, as reflected by diminished frequency and duration of symptoms and improved quality of life. The therapy is considered safe, with side effects limited mostly to minor local reactions, and only occasional cases of systemic adverse reactions. PMID- 21621059 TI - Sublingual immunotherapy. AB - Sublingual immunotherapy (SLIT) has been shown to be safe and efficacious in treating allergic rhinitis. It has been used in Europe for more than 20 years, and interest in the United States is increasing. SLIT has been shown to elicit immunologic changes similar to subcutaneous injection immunotherapy. SLIT may prevent new sensitizations, improve asthma control, and decrease asthma development in allergic individuals. Although differences in antigen quantification and standardization make European dosing schemes difficult to translate in the United States, several new studies suggest the range for effective dosing. Further studies will help clarify optimal dosing. PMID- 21621060 TI - The allergic march: can we prevent allergies and asthma? AB - The allergic march is a progression of atopic disease from eczema to asthma, and then to allergic rhinoconjunctivitis. It appears to be caused by a regional allergic response with breakdown of the local epithelial barrier that initiates systemic allergic inflammation. Genetic and environmental factors predispose to developing the allergic march. There are data to support 4 possible interventions to prevent the allergic march from progressing to asthma: (1) supplements of dietary probiotics, (2) exclusive breast feeding during the first few months of life, or, alternatively (3) use of extensively hydrolyzed infant formulas, (4) treatment with inhalant allergen immunotherapy by either subcutaneous or sublingual methods. PMID- 21621061 TI - The surgical management of allergic rhinitis. AB - In their discussion of the treatment of allergic rhinitis, the authors present key features of the disease and its management, allergen responses, the role of the inferior turbinate, and reviews of outcomes with submucosal resection, total inferior turbinectomy, cryosurgery, laser cautery, radical turbinectomy, submucous turbinectomy, submucous electrocautery, and microdebriber turbinoplasty. The authors discuss radiofrequency ablation and coblation outcomes and complications, along with the role of endoscopic sinus surgery in allergic rhinitis and emphasize the need for Otolaryngologists to be facile with a variety of procedures for best outcomes. PMID- 21621062 TI - Inhalant allergies in children. AB - Children with chronic or recurrent upper respiratory inflammatory disease (rhinitis) should be considered for inhalant allergies. Risk factors for inhalant allergies in children include a first-degree relative with allergies, food allergy in infancy, and atopic dermatitis. Although inhalant allergies are rare in infancy, inhalant allergies are common in older children and impair quality of life and productivity. Differentiating between viral and allergic rhinitis can be challenging in children, but the child's age, history, and risk factors can provide helpful information. Allergic rhinitis is a risk factor for asthma, and if one is present, medical consideration of the other is warranted. PMID- 21621063 TI - Food allergy in adults and children. AB - Food allergy is defined as an adverse health effect arising from a specific immune response that occurs reproducibly on exposure to a given food and is distinct from food intolerance. Clinical manifestations of food allergy are varied and involve many systems including respiratory, cutaneous, and gastrointestinal. The double-blinded placebo-controlled oral food challenge remains the gold standard for the diagnosis of IgE-mediated food allergy. Areas of ongoing research include improved understanding of determinants for the development of tolerance versus sensitization for foods, the role of diagnostic testing for specific epitopes for food allergens, and the use of oral immunotherapy for IgE-mediated food allergy. PMID- 21621064 TI - Skeletal muscle is a vital tissue. Preface. PMID- 21621065 TI - Origin of vertebrate limb muscle: the role of progenitor and myoblast populations. AB - Muscle development, growth, and regeneration take place throughout vertebrate life. In amniotes, myogenesis takes place in four successive, temporally distinct, although overlapping phases. Understanding how embryonic, fetal, neonatal, and adult muscle are formed from muscle progenitors and committed myoblasts is an area of active research. In this review we examine recent expression, genetic loss-of-function, and genetic lineage studies that have been conducted in the mouse, with a particular focus on limb myogenesis. We synthesize these studies to present a current model of how embryonic, fetal, neonatal, and adult muscle are formed in the limb. PMID- 21621066 TI - Developmental origins of fusion-negative rhabdomyosarcomas. AB - Rhabdomyosarcomas (RMS) are very heterogeneous tumors that can be divided into three major groups: alveolar rhabdomyosarcoma, embryonal rhabdomyosarcoma, and pleomorphic rhabdomyosarcoma. Concerted efforts over the past a decade have led to an understanding of the genetic underpinnings of many human tumors through genetically engineered models; however, left largely behind in this effort have been rare tumors with poorly understood chromosomal abnormalities including the vast majority of RMS lacking a pathognomonic translocation, i.e. fusion-negative RMS. In this chapter, we review the characteristic genetic abnormalities associated with human RMS and the genetically engineered animal models for these fusion-negative RMS. We explore not only how specific combinations of mutations and cell of origin give rise to different histologically and biologically distinguishable pediatric and adult RMS subtypes, but we also examine how tumor cell phenotype (and tumor "stem" cell phenotype) can vary markedly from the cell of origin. PMID- 21621068 TI - NF-kappaB signaling in skeletal muscle health and disease. AB - Muscle development, growth, and maintenance require an intricate and timely series of events initiated through a multitude of signaling pathways. The very nature of skeletal muscle requires tremendous plasticity to accommodate the need for anabolism or catabolism, and deregulation of these processes may be a tipping point in the development or progression of various skeletal muscle disorders. Among the relevant signaling pathways, NF-kappaB has emerged as a critical factor involved in various facets of muscle homeostasis. In this review, we summarize the NF-kappaB signaling pathway and provide a fresh perspective into the regulation and function of this transcription factor, underlying both the physiological and pathophysiological states of skeletal muscle. PMID- 21621069 TI - Blood vessels and the satellite cell niche. AB - The fate of stem cell is regulated by cues received from the surrounding area. Recently, the concept of "stem cell zone"--rather than a predefined niche- introduced the notion of dynamic and permanent interactions between stem cells and their microenvironment. In adult skeletal muscle, satellite cells are considered as the main stem cells responsible for muscle repair and maintenance. They are localized close to vessels regardless their state of activation and differentiation. Moreover, the number of satellite cells is positively correlated to the capillarization of the myofiber. Angiogenesis has been known for a long time to be essential for muscle repair. However, relationships between vessel cells and satellite/myogenic cells that govern myogenic cell expansion, myogenesis, and angiogenesis have been only recently investigated. In this chapter, we discuss the possible existence of a vascular amplifying/differentiating niche, in an attempt to reconciliate several recent observations showing that satellite/myogenic cells interact with various cell types during the time course of muscle regeneration. Indeed, endothelial cells (ECs) stimulate myogenic cell growth and, inversely, differentiating myogenic cells promote angiogenesis. However, stromal cells may also provide some proliferating or differentiating cues to satellite/myogenic cells in this vascular area. Although some molecular effectors have been identified, including growth factors and cytokines, molecular regulations that occur within this vascular amplifying/differentiating niche requires further investigation. At the end of muscle repair, maturation of newly formed vessels takes place. In this context, we discuss the potential quiescence niche of satellite cells and the specific role of periendothelial cells. Indeed, periendothelial cells promote the return to quiescence of a subset of satellite/myogenic cells and maintain their quiescence (through Angiopoietin-1/Tie-2 signaling). We ask to what extent the environment may control the fate choice of satellite/myogenic cells and we also question the "hypoxic niche" in skeletal muscle, such a quiescence niche having being observed in the bone marrow. PMID- 21621067 TI - Sculpting chromatin beyond the double helix: epigenetic control of skeletal myogenesis. AB - Satellite cells (SCs) are the main source of adult skeletal muscle stem cells responsible for muscle growth and regeneration. By interpreting extracellular cues, developmental regulators control quiescence, proliferation, and differentiation of SCs by influencing coordinate gene expression. The scope of this review is limited to the description and discussion of protein complexes that introduce and decode heritable histone and chromatin modifications and how these modifications are relevant for SC biology. PMID- 21621070 TI - Nonmyogenic cells in skeletal muscle regeneration. AB - Although classical dogma dictates that satellite cells are the primary cell type involved in skeletal muscle regeneration, alternative cell types such as a variety of inflammatory and stromal cells are also actively involved in this process. A model describing myogenic cells as direct contributors to regeneration and nonmyogenic cells from other developmental sources as important accessories has emerged, with similar systems having been described in numerous other tissues in the body. Increasing evidence supports the notion that inflammatory cells function as supportive accessory cells, and are not merely involved in clearing damage following skeletal muscle injury. Additionally, recent studies have highlighted the role of tissue resident mesenchymal cell populations as playing a central role in regulating regeneration. These "accessory" cell populations are proposed to influence myogenesis via direct cell contact and secretion of paracrine trophic factors. The basic foundations of accessory cell understanding should be recognized as a crucial component to all prospects of regenerative medicine, and this chapter intends to provide a comprehensive background on the current literature describing immune and tissue-resident mesenchymal cells' role in skeletal muscle regeneration. PMID- 21621071 TI - Cellular and molecular mechanisms regulating fibrosis in skeletal muscle repair and disease. AB - The repair of an injured tissue is a complex biological process involving the coordinated activities of tissue-resident and infiltrating cells in response to local and systemic signals. Following acute tissue injury, inflammatory cell infiltration and activation/proliferation of resident stem cells is the first line of defense to restore tissue homeostasis. However, in the setting of chronic tissue damage, such as in Duchenne Muscular Dystrophy, inflammatory infiltrates persist, the ability of stem cells (satellite cells) is blocked and fibrogenic cells are continuously activated, eventually leading to the conversion of muscle into nonfunctional fibrotic tissue. This review explores our current understanding of the cellular and molecular mechanisms underlying efficient muscle repair that are dysregulated in muscular dystrophy-associated fibrosis and in aging-related muscle dysfunction. PMID- 21621072 TI - Ferlin proteins in myoblast fusion and muscle growth. AB - Myoblast fusion contributes to muscle growth in development and during regeneration of mature muscle. Myoblasts fuse to each other as well as to multinucleate myotubes to enlarge the myofiber. The molecular mechanisms of myoblast fusion are incompletely understood. Adhesion, apposition, and membrane fusion are accompanied by cytoskeletal rearrangements. The ferlin family of proteins is implicated in human muscle disease and has been implicated in fusion events in muscle, including myoblast fusion, vesicle trafficking and membrane repair. Dysferlin was the first mammalian ferlin identified and it is now known that there are six different ferlins. Loss-of-function mutations in the dysferlin gene lead to limb girdle muscular dystrophy and the milder disorder Miyoshi Myopathy. Dysferlin is a membrane-associated protein that has been implicated in resealing disruptions in the muscle plasma membrane. Newer data supports a broader role for dysferlin in intracellular vesicular movement, a process also important for resealing. Myoferlin is highly expressed in myoblasts that undergoing fusion, and the absence of myoferlin leads to impaired myoblast fusion. Myoferlin also regulates intracellular trafficking events, including endocytic recycling, a process where internalized vesicles are returned to the plasma membrane. The trafficking role of ferlin proteins is reviewed herein with a specific focus as to how this machinery alters myogenesis and muscle growth. PMID- 21621073 TI - Circadian rhythms, the molecular clock, and skeletal muscle. AB - Almost all organisms ranging from single cell bacteria to humans exhibit a variety of behavioral, physiological, and biochemical rhythms. In mammals, circadian rhythms control the timing of many physiological processes over a 24-h period, including sleep-wake cycles, body temperature, feeding, and hormone production. This body of research has led to defined characteristics of circadian rhythms based on period length, phase, and amplitude. Underlying circadian behaviors is a molecular clock mechanism found in most, if not all, cell types including skeletal muscle. The mammalian molecular clock is a complex of multiple oscillating networks that are regulated through transcriptional mechanisms, timed protein turnover, and input from small molecules. At this time, very little is known about circadian aspects of skeletal muscle function/metabolism but some progress has been made on understanding the molecular clock in skeletal muscle. The goal of this chapter is to provide the basic terminology and concepts of circadian rhythms with a more detailed review of the current state of knowledge of the molecular clock, with reference to what is known in skeletal muscle. Research has demonstrated that the molecular clock is active in skeletal muscles and that the muscle-specific transcription factor, MyoD, is a direct target of the molecular clock. Skeletal muscle of clock-compromised mice, Bmal1(-/-) and Clock(Delta19) mice, are weak and exhibit significant disruptions in expression of many genes required for adult muscle structure and metabolism. We suggest that the interaction between the molecular clock, MyoD, and metabolic factors, such as PGC-1, provide a potential system of feedback loops that may be critical for both maintenance and adaptation of skeletal muscle. PMID- 21621075 TI - The urologic clinics of North America. Erectile dysfunction. PMID- 21621074 TI - Regulation of nucleocytoplasmic transport in skeletal muscle. AB - Proper skeletal muscle function is dependent on spatial and temporal control of gene expression in multinucleated myofibers. In addition, satellite cells, which are tissue-specific stem cells that contribute critically to repair and maintenance of skeletal muscle, are also required for normal muscle physiology. Gene expression in both myofibers and satellite cells is dependent upon nuclear proteins that require facilitated nuclear transport. A unique challenge for myofibers is controlling the transcriptional activity of hundreds of nuclei in a common cytoplasm yet achieving nuclear selectivity in transcription at specific locations such as neuromuscular synapses and myotendinous junctions. Nucleocytoplasmic transport of macromolecular cargoes is regulated by a complex interplay among various components of the nuclear transport machinery, namely nuclear pore complexes, nuclear envelope proteins, and various soluble transport receptors. The focus of this review is to highlight what is known about the nuclear transport machinery and its regulation in skeletal muscle and to consider the unique challenges that multinucleated muscle cells as well as satellite cells encounter in regulating nucleocytoplasmic transport during cell differentiation and tissue adaptation. Understanding how regulated nucleocytoplasmic transport controls gene expression in skeletal muscle may lead to further insights into the mechanisms contributing to muscle growth and maintenance throughout the lifespan of an individual. PMID- 21621076 TI - Recovery of erectile function after robotic prostatectomy: evidence-based outcomes. AB - Several reported advantages of the robotic-assisted laparoscopic approach to the treatment of clinically localized prostate cancer include superior results for erectile function as one of the critical outcomes of radical prostate surgery. This article provides a critical assessment of the evidence that exists for erectile function outcomes based on a systematic literature review. We found that the low methodological and reporting quality of existing studies did not appear well suited to guide clinical practice. A new framework of prospective investigation using validated patient self-assessment instruments would seem critical to the future advancement of this field. PMID- 21621077 TI - Penile rehabilitation after prostate cancer treatment: outcomes and practical algorithm. AB - The number of patients diagnosed with prostate cancer was estimated to be 192,000 in 2009 according to the American Cancer Society. The prevalence of reported erectile dysfunction after radical prostatectomy has significant variance. Among the studies in which the nerve-sparing status was described, erectile function recovery adequate for sexual intercourse was achieved in 50% of patients. This article reviews the animal and human studies in this field and provides a useful penile rehabilitation algorithm. PMID- 21621078 TI - Testosterone and prostate cancer: what are the risks for middle-aged men? AB - With increased recognition of the benefits of testosterone (T) therapy for middle aged men, there has been a concomitant reexamination of the historical fear that raising T will result in more prostate cancer (PCa). Studies have failed to show increased risk of PCa in men with higher serum T, and supraphysiologic T fails to increase prostate volume or prostate-specific antigen in healthy men. This apparent paradox is explained by the Saturation Model, which posits a finite capacity of androgen to stimulate PCa growth. Modern studies indicate no increased risk of PCa among men with serum T in the therapeutic range. PMID- 21621079 TI - Erectile dysfunction and depression: screening and treatment. AB - The comorbid conditions erectile dysfunction (ED) and depression are highly prevalent in men. Multiple regression analysis to control for all other predictors of ED indicate that men with high depression scores are nearly twice as likely to report ED than nondepressed men. Depression continues to be among the most common comorbid problems in men with ED, both in the community and in clinical samples. This article reviews the current knowledge about the relationship between ED and depression, the effect of treatments for depression on ED, ways to improve screening for depression, and treatment of ED in patients with this comorbidity. PMID- 21621080 TI - Psychological factors associated with male sexual dysfunction: screening and treatment for the urologist. AB - Male sexual dysfunctions, including erectile dysfunction, hypoactive sexual desire disorder, premature ejaculation, and delayed ejaculation, are a complex amalgam of interrelated biological, psychological, and contextual variables that can combine to produce distressing symptoms both for the male diagnosed with the dysfunction and for his partner. This article describes the assessment process for identifying the psychological concerns associated with the man's sexual complaint, and presents a stepwise algorithm for treating mild to moderate psychosocial issues. Physicians' awareness of psychological and interpersonal issues will help them better manage patients' ongoing medical treatment and limit discontinuation of efficacious therapies. PMID- 21621081 TI - Doppler blood flow analysis of erectile function: who, when, and how. AB - The underlying processes in vasculogenic erectile dysfunction (ED) are arterial insufficiency, venoocclusive disease, or combinations of both. Doppler blood flow analysis is a diagnostic modality useful in elucidating the cause of ED and the magnitude of its severity. This article describes the procedural techniques, typical findings, and relevant pathophysiology for in-office Doppler studies. Specific conditions include arterial insufficiency, venous occlusive disease, Peyronie's disease, and priapism. PMID- 21621082 TI - Newer phosphodiesterase inhibitors: comparison with established agents. AB - Erectile dysfunction is defined as the consistent or recurrent inability to attain or maintain penile erection sufficient for sexual performance. Self reported erectile dysfunction has increased significantly as men seek effective therapy, such as oral phosphodiesterase 5 inhibitors (PDE5i). PDE5i are now the drugs of choice in the initial therapy of erectile dysfunction. This review compares the currently available PDE5i with the second-generation PDE5i, which are soon to be available. PMID- 21621083 TI - Central nervous system agents and erectile dysfunction. AB - Several centrally acting agents have shown potential to improve erectile function in men with ED. They still lack adequate data in efficacy and tolerability. Nasal formulations of apomorphine and bremelanotide seem to be the most likely candidates for future approval. They may play a role, specifically in men who fail phosphodiesterase 5 (PDE5) therapy, are unable to take PDE5 inhibitors because of side effects, or are on nitrate therapy. This article reviews the centrally acting agents and the data on their efficacy. PMID- 21621084 TI - Testosterone deficiency and risk factors in the metabolic syndrome: implications for erectile dysfunction. AB - The most common cause of erectile dysfunction (ED) is penile vascular insufficiency. This is usually part of a generalized endothelial dysfunction and is related to several conditions, including type 2 diabetes mellitus, hypertension, hyperlipidemia, and obesity. These conditions underlie the pathophysiology of metabolic syndrome (MetS). Hypogonadism, or testosterone deficiency (TD), is an integral component of the pathology underlying endothelial dysfunction and MetS, with insulin resistance (IR) at its core. Testosterone replacement therapy for TD has been shown to ameliorate some of the components of the MetS, improve IR, and may serve as treatment for decreasing cardiovascular and ED risk. PMID- 21621085 TI - Priapism: new concepts in medical and surgical management. AB - Advances have recently been made in both medical and surgical management of priapism, and these offer improvements in the level of care afforded such patients. Further developments can be expected based on ongoing progress, particularly in the area of molecular science, which is the primary source for driving novel therapeutic approaches. Continued action to address the health care administrative concerns of those most commonly affected by priapism, specifically individuals with sickle cell disease, is also appropriate. All successes in these arenas ensure that afflicted individuals avoid the health burdens of priapism and preserve sexual function. PMID- 21621086 TI - Peyronie's disease: review of nonsurgical treatment options. AB - The purpose of this article is to review the contemporary literature on nonsurgical therapies for Peyronie's disease (PD); focus on randomized, placebo controlled trials; and review the latest guidelines for the management of PD from the International Consultation on Sexual Medicine. A combination of oral agents or intralesional injection with traction therapy may provide a synergy between the chemical effects of the drugs and the mechanical effects of traction. Until a reliable treatment emerges, some of the nonsurgical treatments discussed can be used to stabilize the scarring process and may result in some reduction of deformity with improved sexual function. PMID- 21621087 TI - Peyronie disease: plication or grafting. AB - Peyronie disease (PD) is an incurable, sexually debilitating disease resulting in penile deformity, coital failure, and significant psychological stress for patients and their partners. Appropriate treatment should be individualized and tailored to the patient's goals and expectations, disease history, physical examination findings, and erectile function. After medical therapy is considered and the disease has stabilized, surgical correction, including tunical shortening or lengthening procedures, is an excellent option for patients with functional impairment caused by PD. Outcomes are satisfactory when proper treatment decisions are made, with the goal being expected return to normal sexual function following PD treatment. PMID- 21621088 TI - Penile prosthesis implantation in the era of medical treatment for erectile dysfunction. AB - Penile prosthesis implantation, the oldest of the modern treatments for erectile dysfunction (ED), still plays an important role despite the advent of less invasive alternatives. For some men with ED, penile prosthesis implantation is the only effective or acceptable treatment. Penile prosthesis implantation remains a viable option in the contemporary management of ED as evidenced by annual penile prosthesis implantation cases in the United States rising from 17,540 in 2000 to 22,420 in 2009. Improvements in prosthesis design and implantation techniques have resulted in significant increases in device survival and patient satisfaction. PMID- 21621089 TI - Penile prosthesis infection: approaches to prevention and treatment. AB - Epidemiologic studies have estimated that more than 50% of men ages 40 to 70 have some form of erectile dysfunction. Penile prosthesis implantation remains a mainstay for treatment of erectile dysfunction unresponsive to other less invasive methods. Improvements in penile prosthesis design have extended the long term survival of implants. As the improved design of prostheses has led to their increased mechanical survival, other complications, such as infection, have emerged as the leading causes of implant failure. This article focuses on approaches to prevention and treatment of penile prosthesis infection. PMID- 21621090 TI - Images in emergency medicine. Woman with left wrist pain. Aneurysmal bone cyst. PMID- 21621091 TI - Rethinking testing for pulmonary embolism: less is more. PMID- 21621092 TI - Critical issues in the evaluation and management of adult patients presenting to the emergency department with suspected pulmonary embolism. AB - This clinical policy from the American College of Emergency Physicians is the revision of a 2003 clinical policy on the evaluation and management of adult patients presenting with suspected pulmonary embolism (PE).(1) A writing subcommittee reviewed the literature to derive evidence-based recommendations to help clinicians answer the following critical questions: (1) Do objective criteria provide improved risk stratification over gestalt clinical assessment in the evaluation of patients with possible PE? (2) What is the utility of the Pulmonary Embolism Rule-out Criteria (PERC) in the evaluation of patients with suspected PE? (3)What is the role of quantitative D-dimer testing in the exclusion of PE? (4) What is the role of computed tomography pulmonary angiogram of the chest as the sole diagnostic test in the exclusion of PE? (5) What is the role of venous imaging in the evaluation of patients with suspected PE? (6) What are the indications for thrombolytic therapy in patients with PE? Evidence was graded and recommendations were given based on the strength of the available data in the medical literature. PMID- 21621093 TI - A systematic review and qualitative analysis to inform the development of a new emergency department-based geriatric case management model. AB - STUDY OBJECTIVE: We inform the future development of a new geriatric emergency management practice model. We perform a systematic review of the existing evidence for emergency department (ED)-based case management models designed to improve the health, social, and health service utilization outcomes for noninstitutionalized older patients within the context of an index ED visit. METHODS: This was a systematic review of English-language articles indexed in MEDLINE and CINAHL (1966 to 2010), describing ED-based case management models for older adults. Bibliographies of the retrieved articles were reviewed to identify additional references. A systematic qualitative case study analytic approach was used to identify the core operational components and outcome measures of the described clinical interventions. The authors of the included studies were also invited to verify our interpretations of their work. The determined patterns of component adherence were then used to postulate the relative importance and effect of the presence or absence of a particular component in influencing the overall effectiveness of their respective interventions. RESULTS: Eighteen of 352 studies (reported in 20 articles) met study criteria. Qualitative analyses identified 28 outcome measures and 8 distinct model characteristic components that included having an evidence-based practice model, nursing clinical involvement or leadership, high-risk screening processes, focused geriatric assessments, the initiation of care and disposition planning in the ED, interprofessional and capacity-building work practices, post-ED discharge follow up with patients, and evaluation and monitoring processes. Of the 15 positive study results, 6 had all 8 characteristic components and 9 were found to be lacking at least 1 component. Two studies with positive results lacked 2 characteristic components and none lacked more than 2 components. Of the 3 studies with negative results demonstrating no positive effects based on any outcome tested, one lacked 2, one lacked 3, and one lacked 4 of the 8 model components. CONCLUSION: Successful models of ED-based case management models for older adults share certain key characteristics. This study builds on the emerging literature in this area and leverages the differences in these models and their associated outcomes to support the development of an evidence-based normative and effective geriatric emergency management practice model designed to address the special care needs and thereby improve the health and health service utilization outcomes of older patients. PMID- 21621094 TI - Commentary: using NOPUS data to identify at-risk groups. PMID- 21621095 TI - Firearm injury prevention. Policy statement. PMID- 21621096 TI - Change of shift. Of masses and men. PMID- 21621097 TI - Annals of emergency medicine journal club. To shock or not to shock: that is the question; is there an answer? Answers to the January 2011 journal club questions. PMID- 21621098 TI - Images in emergency medicine. Woman with right eye pain and swelling. Partial globe rupture. PMID- 21621099 TI - CMS selects an invalid imaging measure: deja vu all over again. PMID- 21621100 TI - Etomidate, sepsis, and informed consent. PMID- 21621102 TI - [Imaging and postoccupational surveillance after exposure to asbestos: new recommendations]. PMID- 21621103 TI - [Evaluation of glomerular filtration rate with magnetic resonance imaging]. AB - Glomerular Filtration Rate (GFR) is one of the cardinal indices of renal function and is used clinically as the gold standard of renal dysfunction. In the past decade, many studies using dynamic contrast-enhanced MRI (DCE MRI) to measure GFR have been published. The MRI evaluation of GFR centers on visualizing the passage of contrast material (Gadolinium chelates) through the kidney. MRI appears as a promising tool but still relatively difficult to implement in the assessment of GFR. A high heterogeneity of protocols (e.g., in acquisition mode, dose of contrast, postprocessing techniques) is noted in the literature, reflecting the number of technical challenges that should first be solved in order to reach a consensus, and the reported accuracy and reproducibility are insufficient for justifying their use in clinical practice now. This paper presents and discusses the different steps that can be used to quantify the GFR by MRI. PMID- 21621104 TI - [Peritoneal parasitic teratoma and chemical dermoid peritonitis]. AB - Chemical peritonitis occurs following intraperitoneal rupture of a mature ovarian dermoid. Rupture may be acute and spontaneous, typically during pregnancy, or iatrogenic. Low grade ruptures lead to parasitic peritoneal dermoid cysts, usually involving the greater omentum, cul-de-sac of Douglas and perihepatic region. Radiologists should be familiar with their appearance to correctly diagnose the condition and not mistake the disease for peritoneal carcinomatosis. PMID- 21621105 TI - [Radiology as seen by medical students]. AB - PURPOSE: To determine the impact of exposing medical students to medical imaging during the first year of the second cycle of medical school (DCEM1) on their perception of this medical specialty and the acquisition of its basic concepts. MATERIALS AND METHODS: All students in the 2007-2008 graduation class entering into the first year of the second cycle of medical school were anonymously enrolled into this project that included pre-rotation and post-rotation questionnaires, theory classes followed by clinical rotations with clearly predetermined objectives. RESULTS: A total of 108 students were enrolled, with 70% being females. The parents of the students had a medical or paramedical profession in 46.3% of cases. Before the rotation, 61.6% of students perceived a difference between a hospital-based practice and private practice. Fifty-two percent of students had a clear idea of their professional future prior to the rotation. Five students (4.7%) believed prior to the rotation that it might have an impact on their professional future, versus 63% after the rotation (P<0.0001). The students whose parents work in the medical or paramedical field do not have a better defined idea of their professional future; on the other hand, they have more interest for radiology (73.6% with high or very high interest versus 52.8%, P=0.03). After the rotation, there was a significant increase in the number of students with high or very high interest for radiology (77.8% versus 66.7%, P=0.023). A student noted that he would redirect his career to radiology. There was also a significant increase in the number of students perceiving a difference between a hospital-based practice and private practice (82.2% versus 61.6%, P=0.003). With regards to radiology knowledge before and after the rotation, there was a significant increase of mean scores (P<0.001). Eighty-eight percent of students were satisfied or very satisfied with the radiology rotation. Overall, the students believe that 70% of the objectives were achieved. The only criticism from the students was that the clinical instructors were overworked. CONCLUSION: Early exposure of medical students to radiology increases their level of interest for the specialty and increases the perception of differences between a hospital-based practice and private practice. The overall knowledge of students about radiology was improved, but the workload of clinical instructors impaired the quality of the rotation. PMID- 21621106 TI - [Accuracy of CT-guided chest biopsy for benign lesions]. AB - PURPOSE: To demonstrate that CT-guided chest biopsy can effectively prevent surgery for patients with benign tumors. PATIENTS AND METHODS: We present a cohort of patients who underwent CT-guided biopsy for a chest lesion where diagnosis could not be achieved through other means. The gold standard was defined, based on availability, by surgery or imaging and clinical follow-up over a 2-year period. RESULTS: From a total of 114 biopsied lesions, 101 were malignant and 13 were benign. Sensitivity and specificity values for benign lesions were 92.1 and 92.3% respectively with PPV and NPV of 60 and 98.9%. Sensitivity and specificity values for malignant lesions were 86.1 and 100% respectively with PPV and NPV of 100 and 48%. No statistically significant correlation could be established between the groups with concordant and discordant results. CONCLUSION: The diagnosis of specific benign lesion on CT guided biopsy, all sizes and sites included, may effectively prevent unnecessary surgery. PMID- 21621107 TI - [Evaluation of suspicious nipple discharge with MRI: review of 50 cases]. AB - PURPOSE: To determine the role of MRI in the evaluation and management of patients with suspicious nipple discharge and normal mammographic and US evaluation. PATIENTS AND METHODS: A total of 50 patients with suspicious nipple discharge and normal mammographic and US evaluation prospectively underwent MRI. The first 16 patients underwent routine breast MRI, while MR-ductography with image fusion at the console was added for the last 34 patients. RESULTS: In 22 of 25 high-risk and malignant lesions, MRI showed enhancement whereas it was normal in three cases. In 25 benign cases (resolution of discharge/benign non proliferative breast disease), MRI was negative in 22 cases and falsely positive in three cases. CONCLUSION: In this clinical setting, MRI shows excellent sensitivity, PPV and NPV. A negative result on MRI would support clinical follow up as opposed to surgery. PMID- 21621108 TI - [Transient elastography of calcaneal tendon: preliminary results and future prospects]. AB - PURPOSE: To evaluate transient ShearWave elastography of the normal Achilles' tendon. PATIENTS AND METHODS: The Achilles' tendon of 30 normal subjects were prospectively assessed using a Rubi V1Sq prototype (Supersonic Imagine). Quantitative elastography maps displayed in kilopascals with a scale of 0 to 600kPa were generated from transverse and longitudinal images at 3 different levels of plantar flexion. Subgroups were compared and analyzed based on proven or suspected variation factors (age, gender, level of physical activity). RESULTS: On sagittal images, mean elasticity was 104+/-46kPa during extension, 464+/-144kPa in neutral position and 410+/-196kPa during maximum dorsiflexion. There was significant increase in elasticity when the Achilles' tendon was maximally stretched (p<0.01). The intra-individual correlation between right and left tendons during ankle extension was good (Pearson Coefficient 0.8; p<0.01). Finally, physically active subjects showed significantly greater elasticity than non-active subjects (p<0.05). CONCLUSION: Transient ShearWave elastography of the Achilles' tendon is a simple technique that provides real-time information about tissue elasticity. PMID- 21621109 TI - [The current asbestos scandal]. PMID- 21621110 TI - [Prenatal helical CT diagnosis of severe fibular hemimelia]. PMID- 21621111 TI - [Intussusception on multiple metastases of malignant melanoma]. PMID- 21621112 TI - [Hemangioma of the breast: mammographic, sonographic and MRI features]. PMID- 21621113 TI - [Postmortem CT-angiography: feasibility of US-guided vascular access]. PMID- 21621114 TI - [Exposure to asbestos : radiography and chest CT]. PMID- 21621115 TI - [Post-professional follow-up after asbestos exposure. Guidelines of the audition commission]. PMID- 21621116 TI - [A case of cherubism. Answer to March e-quid]. PMID- 21621118 TI - Will there be a future role for radiation in the neo-adjuvant therapy for rectal cancer? PMID- 21621119 TI - Expression of excision repair cross-complementation group 1 as predictive marker for nasopharyngeal cancer treated with concurrent chemoradiotherapy. AB - PURPOSE: Cisplatin-based concurrent chemoradiotherapy is the standard treatment of nasopharyngeal cancer. The expression of excision repair cross-complementation group 1 (ERCC1) has been reported to be associated with resistance to platinum based chemotherapy. We evaluated whether ERCC1 expression could predict the treatment response and survival outcome of patients with locally advanced nasopharyngeal cancer who were treated with cisplatin-based concurrent chemoradiotherapy. METHODS AND MATERIALS: Immunohistochemistry was used to examine the expression of ERCC1 in nasopharyngeal tumor tissue. Patients were categorized into either a resistant or sensitive group depending on their treatment response outcome. A total of 77 patients were assessed in the present study. RESULTS: The resistant and sensitive groups included 25 and 52 patients, respectively. ERCC1 expression was positive in the tumor tissue for 39 of the 77 patients (51%). Significantly more ERCC1-negative tumors were in the sensitive group than in the resistant group (p = .035). In terms of survival outcome, univariate analysis determined that patients with ERCC1-negative tumors had longer disease-free survival (p = .076) and overall survival (p = .013) than patients with ERCC1-positive tumors. Multivariate analysis determined that negative ERCC expression in tumors was an independent predictor for prolonged overall survival (hazard ratio, 0.14; 95% confidence interval, 0.03-0.71). CONCLUSION: These results suggest that ERCC1 expression might be a useful predictive marker in patients with locally advanced nasopharyngeal cancer who are under consideration for cisplatin-based concurrent chemoradiotherapy. PMID- 21621120 TI - Positive surgical margins in soft tissue sarcoma treated with preoperative radiation: is a postoperative boost necessary: in regard to Al Yami et al. (Int J Radiat Oncol Biol Phys 2010;77:1191-1197). PMID- 21621123 TI - In regard to "Radiation-induced liver fibrosis is mitigated by gene therapy inhibiting transforming growth factor-beta signalling in the rat" by Shi-Suo Du et al. (Int J Radiat Oncol Biol Phys 2010;78:1513-1523). PMID- 21621124 TI - Imaging work up of small aggressive lymph nodes: in regard to Ghadjar et al. (Int J Radiat Oncol Biol Phys 2010;78:1366-1372). PMID- 21621127 TI - Palliative sedation in Flemish palliative care units. PMID- 21621129 TI - Death service ratio: a measure of hospice utilization and cost impact. PMID- 21621130 TI - Studies comparing Numerical Rating Scales, Verbal Rating Scales, and Visual Analogue Scales for assessment of pain intensity in adults: a systematic literature review. AB - CONTEXT: The use of unidimensional pain scales such as the Numerical Rating Scale (NRS), Verbal Rating Scale (VRS), or Visual Analogue Scale (VAS) is recommended for assessment of pain intensity (PI). A literature review of studies specifically comparing the NRS, VRS, and/or VAS for unidimensional self-report of PI was performed as part of the work of the European Palliative Care Research Collaborative on pain assessment. OBJECTIVES: To investigate the use and performance of unidimensional pain scales, with specific emphasis on the NRSs. METHODS: A systematic search was performed, including citations through April 2010. All abstracts were evaluated by two persons according to specified criteria. RESULTS: Fifty-four of 239 papers were included. Postoperative PI was most frequently studied; six studies were in cancer. Eight versions of the NRS (NRS-6 to NRS-101) were used in 37 studies; a total of 41 NRSs were tested. Twenty-four different descriptors (15 for the NRSs) were used to anchor the extremes. When compared with the VAS and VRS, NRSs had better compliance in 15 of 19 studies reporting this, and were the recommended tool in 11 studies on the basis of higher compliance rates, better responsiveness and ease of use, and good applicability relative to VAS/VRS. Twenty-nine studies gave no preference. Many studies showed wide distributions of NRS scores within each category of the VRSs. Overall, NRS and VAS scores corresponded, with a few exceptions of systematically higher VAS scores. CONCLUSION: NRSs are applicable for unidimensional assessment of PI in most settings. Whether the variability in anchors and response options directly influences the numerical scores needs to be empirically tested. This will aid in the work toward a consensus-based, standardized measure. PMID- 21621131 TI - The ethics of pacemaker deactivation in terminally ill patients. AB - A core principle of American medical ethics holds that an informed and capacitated patient has the right to have treatments withdrawn or withheld. Nevertheless, many clinicians remain reluctant to honor a request to deactivate a patient's pacemaker. This article describes a case in which a patient was denied her request for pacemaker deactivation. Several reasons for this reluctance are discussed, including historical, practical, and ethical considerations for opposing pacemaker deactivation. Ultimately, however, from an ethical standpoint, pacemaker deactivation is similar to withdrawal of other therapies. Fortunately, a recent expert consensus statement supports a patient's right to have her pacemaker deactivated. Pacemaker deactivation should only be performed after robust informed consent, which must include discussion of risks, benefits, and all viable alternatives based on the patient's values and goals. PMID- 21621134 TI - Prospective follow-up studies of ADHD: helping establish a valid diagnosis in adults. PMID- 21621135 TI - Hypermentalization in adolescents with borderline personality traits: extending the conceptual framework to younger ages. PMID- 21621136 TI - Monozygotic twins discordant for being bullied: a step closer to understanding the biology of victimization. PMID- 21621137 TI - Criteria, categories, and continua: autism and related disorders in DSM-5. PMID- 21621138 TI - Does ADHD predict substance-use disorders? A 10-year follow-up study of young adults with ADHD. AB - OBJECTIVE: High rates of substance-use disorders (SUD) have been found in samples of adolescents and adults with attention-deficit/hyperactivity disorder (ADHD). Predictors of SUD in children with ADHD who are at risk for the development of SUDs remain understudied. The main aims of this study were to identify clinically meaningful characteristics of children that predicted the future development of SUDs and to see whether the role of these characteristics varied by sex. METHOD: Subjects were children and adolescents with (n = 268; mean age +/- standard deviation = 10.9 +/- 3.2 years) and without (n = 229; mean age 11.9 +/- 3.3 years) DSM-III-R ADHD followed prospectively and blindly over a 10-year follow-up period onto young adult years. Subjects were assessed with structured diagnostic interviews for psychopathology and SUDs. RESULTS: Over the 10-year follow-up period, ADHD was found to be a significant predictor of any SUD (hazards ratio 1.47; 95% confidence interval 1.07-2.02; p = .01) and cigarette smoking (2.38; 1.61-3.53; p < .01). Within ADHD, comorbid conduct disorder (2.74; 1.66-4.52; p < .01) and oppositional defiant disorder (2.21; 1.40-3.51; p < .01) at baseline were also found to be significant predictors of SUDs. Similar results were found for cigarette-, alcohol-, and drug-use disorders. There were few meaningful sex interaction effects. No clinically significant associations were found for any social or family environment factors or for cognitive functioning factors (p > .05 for all comparisons). CONCLUSIONS: These results indicate that ADHD is a significant risk factor for the development of SUDs and cigarette smoking in both sexes. PMID- 21621139 TI - Short-term persistence of DSM-IV ADHD diagnoses: influence of context, age, and gender. AB - OBJECTIVE: Little is known about the effect of social context and gender on persistence of attention-deficit/hyperactivity disorder (ADHD) in children of early and middle school years. The study compared persistence of DSM-IV ADHD and ADHD not otherwise specified (NOS) over 2 years in two groups of Puerto Rican children. METHOD: A three-wave study obtained data on Puerto Rican children 5 through 13 years of age at baseline. Samples were drawn in the South Bronx in New York (n = 1,138) and two metropolitan areas in Puerto Rico (n = 1,353). The Diagnostic Interview Schedule for Children Version IV was used to diagnose ADHD and ADHD-NOS. RESULTS: ADHD or ADHD-NOS diagnosis at wave 1 strongly predicted disorder at waves 2 and 3. ADHD had a significantly stronger predictive effect than ADHD-NOS consistently across site and gender. There was a significant interaction with baseline age. For those younger at baseline, the strength of the prediction of ADHD-NOS was relatively weak; for older children, the presence of ADHD-NOS at baseline predicted risk of subsequent ADHD or ADHD-NOS. CONCLUSIONS: Persistence of ADHD in children of similar ethnicity does not manifest differently across context and gender. Results suggest that age-specific symptom criteria and modification of age-of-onset criteria should be considered for the diagnosis. PMID- 21621140 TI - Theory of mind and emotion regulation difficulties in adolescents with borderline traits. AB - OBJECTIVE: Dysfunctions in both emotion regulation and social cognition (understanding behavior in mental state terms, theory of mind or mentalizing) have been proposed as explanations for disturbances of interpersonal behavior in borderline personality disorder (BPD). This study aimed to examine mentalizing in adolescents with emerging BPD from a dimensional and categorical point of view, controlling for gender, age, Axis I and Axis II symptoms, and to explore the mediating role of emotion regulation in the relation between theory of mind and borderline traits. METHOD: The newly developed Movie for the Assessment of Social Cognition (MASC) was administered alongside self-report measures of emotion regulation and psychopathology to 111 adolescent inpatients between the ages of 12 to 17 (mean age = 15.5 years; SD = 1.44 years). For categorical analyses borderline diagnosis was determined through semi-structured clinical interview, which showed that 23% of the sample met criteria for BPD. RESULTS: Findings suggest a relationship between borderline traits and "hypermentalizing" (excessive, inaccurate mentalizing) independent of age, gender, externalizing, internalizing and psychopathy symptoms. The relation between hypermentalizing and BPD traits was partially mediated by difficulties in emotion regulation, accounting for 43.5% of the hypermentalizing to BPD path. CONCLUSIONS: Results suggest that in adolescents with borderline personality features the loss of mentalization is more apparent in the emergence of unusual alternative strategies (hypermentalizing) than in the loss of the capacity per se (no mentalizing or undermentalizing). Moreover, for the first time, empirical evidence is provided to support the notion that mentalizing exerts its influence on borderline traits through the mediating role of emotion dysregulation. PMID- 21621141 TI - A discordant monozygotic twin design shows blunted cortisol reactivity among bullied children. AB - OBJECTIVE: Childhood adverse experiences are known to engender persistent changes in stress-related systems and brain structures involved in mood, cognition, and behavior in animal models. Uncertainty remains about the causal effect of early stressful experiences on physiological response to stress in human beings, as the impact of these experiences has rarely been investigated while controlling for both genetic and shared environmental influences. METHOD: We tested whether bullying victimization, a repeated adverse experience in childhood, influences cortisol responses to a psychosocial stress test (PST) using a discordant monozygotic (MZ) twin design. Thirty pairs (43.3% males) of 12-year-old MZ twins discordant for bullying victimization were identified in the Environmental Risk (E-Risk) Longitudinal Twin Study, a nationally representative 1994-1995 cohort of families with twins. RESULTS: Bullied and nonbullied MZ twins showed distinct patterns of cortisol secretion after the PST. Specifically, bullied twins exhibited a blunted cortisol response compared with their nonbullied MZ co-twins, who showed the expected increase. This difference in cortisol response to stress could not be attributed to children's genetic makeup, their familial environments, pre-existing and concomitant individual factors, or the perception of stress and emotional response to the PST. CONCLUSION: Results from this natural experiment provide support for a causal effect of adverse childhood experiences on the neuroendocrine response to stress. PMID- 21621142 TI - Autism spectrum disorders according to DSM-IV-TR and comparison with DSM-5 draft criteria: an epidemiological study. AB - OBJECTIVE: The latest definitions of autism spectrum disorders (ASDs) were specified in DSM-IV-TR in 2000. DSM-5 criteria are planned for 2013. Here, we estimated the prevalence of ASDs and autism according to DSM-IV-TR, clarified confusion concerning diagnostic criteria, and evaluated DSM-5 draft criteria for ASD posted by the American Psychiatry Association (APA) in February 2010. METHOD: This was an epidemiological study of 5,484 eight-year-old children in Finland, 4,422 (81%) of them rated via the Autism Spectrum Screening Questionnaire by parents and/or teachers, and 110 examined by using a structured interview, semi structured observation, IQ measurement, school-day observation, and patient records. Diagnoses were assigned according to DSM-IV-TR criteria and DSM-5 draft criteria in children with a full-scale IQ (FSIQ) >=50. Patient records were evaluated in children with an FSIQ <50 to discover diagnoses of ASDs. RESULTS: The prevalence of ASDs was 8.4 in 1,000 and that of autism 4.1 in 1,000 according to DSM-IV-TR. Of the subjects with ASDs and autism, 65% and 61% were high functioning (FSIQ >=70), respectively. The prevalence of pervasive developmental disorder not otherwise specified was not estimated because of inconsistency in DSM-IV-TR criteria. DSM-5 draft criteria were shown to be less sensitive in regard to identification of subjects with ASDs, particularly those with Asperger's syndrome and some high-functioning subjects with autism. CONCLUSIONS: DSM-IV-TR helps with the definition of ASDs only up to a point. We suggest modifications to five details of DSM-5 draft criteria posted by the APA in February 2010. Completing revision of DSM criteria for ASDs is a challenging task. PMID- 21621143 TI - Neuroanatomical and neuropsychological correlates of the cerebellum in children with attention-deficit/hyperactivity disorder--combined type. AB - OBJECTIVE: Studies of healthy individuals and those with cerebellar damage have implicated the cerebellum in a variety of cognitive and behavioral processes. Decreased cerebellar volume has been found in children with attention deficit/hyperactivity disorder (ADHD) and differentially related to behavioral outcomes. The present study investigated whether smaller cerebellar vermis volume was present in children with ADHD-combined type (ADHD-C) compared with controls and whether volume related to parent- and teacher-reported levels of ADHD symptomatology. METHOD: T1-weighted magnetic resonance images and parent- and teacher-reported ADHD symptoms were acquired for 32 children diagnosed with ADHD C and 15 typically developing controls. Participants were right-handed, had no comorbid diagnoses of learning disabilities, conduct disorder, or affective/mood disorder, and were 9 to 15 years of age. RESULTS: Participants with ADHD-C showed significantly smaller volume in the posterior inferior vermis compared with controls. No statistically significant differences were observed for cerebral volume, anterior vermis volume, posterior superior volume, or total cerebellar volume. Regression analyses indicated that a significant amount of the variance in parent-reported Behavior Assessment System for Children, Second Edition, Hyperactivity and Attention and Conners Restless/Impulsive ratings was explained by volume of the posterior inferior vermis. CONCLUSIONS: Consistent with previous studies, children with ADHD had smaller volume in the posterior inferior vermis. New findings emerged with smaller volume of the posterior inferior vermis predicting a significant amount of the variance in parent-reported hyperactivity, attention, and restlessness/impulsivity. Thus, symptoms of hyperactivity and inattention in ADHD may be partly explained by smaller volume of the cerebellar vermis and its connections within the cerebrum. PMID- 21621144 TI - Dissociable patterns of neural activity during response inhibition in depressed adolescents with and without suicidal behavior. AB - OBJECTIVES: Impaired attentional control and behavioral control are implicated in adult suicidal behavior. Little is known about the functional integrity of neural circuitry supporting these processes in suicidal behavior in adolescence. METHOD: Functional magnetic resonance imaging was used in 15 adolescent suicide attempters with a history of major depressive disorder (ATTs), 15 adolescents with a history of depressive disorder but no suicide attempt (NATs), and 14 healthy controls (HCs) during the performance of a well-validated go-no-go response inhibition and motor control task that measures attentional and behavioral control and has been shown to activate prefrontal, anterior cingulate, and parietal cortical circuitries. Questionnaires assessed symptoms and standardized interviews characterized suicide attempts. RESULTS: A 3 group by 2 condition (go-no-go response inhibition versus go motor control blocks) block design whole-brain analysis (p < .05, corrected) showed that NATs showed greater activity than ATTs in the right anterior cingulate gyrus (p = .008), and that NATs, but not ATTs, showed significantly greater activity than HCs in the left insula (p = .004) to go-no-go response inhibition blocks. CONCLUSIONS: Although ATTs did not show differential patterns of neural activity from HCs during the go no-go response inhibition blocks, ATTs and NATs showed differential activation of the right anterior cingulate gyrus during response inhibition. These findings indicate that suicide attempts during adolescence are not associated with abnormal activity in response inhibition neural circuitry. The differential patterns of activity in response inhibition neural circuitry in ATTs and NATs, however, suggest different neural mechanisms for suicide attempt versus major depressive disorder in general in adolescence that should be a focus of further study. PMID- 21621147 TI - XMRV: not a mousy virus. PMID- 21621145 TI - Neurobiology of decision making in depressed adolescents: a functional magnetic resonance imaging study. AB - OBJECTIVE: Despite evidence that impaired reward- and risk-related behavior during adolescence can have potentially serious short- and long-term consequences, few studies have investigated the impact of depression on reward related selection in adolescents. This study examined the relationship between reward-related behavior and prefrontal activations in depressed and healthy adolescents during a decision-making task. METHOD: A total of 22 adolescents with no personal or family history of psychiatric illness and 22 adolescents with major depressive disorder were administered a monetary, two-option decision making task, the Wheel of Fortune, using a functional magnetic resonance imaging protocol. The analysis was focused on the selection phase, i.e., the first phase of the decision-making process, which typically includes two more phases, the anticipation of outcome and the feedback. RESULTS: Similar prefrontal regions were activated in healthy and depressed adolescents during reward-related selection. However, in a contrast involving the selection of high-risk (low probability/high-magnitude reward) versus equal-risk (50% chance of reward) options, healthy adolescents showed greater activation than patients in the right lateral orbitofrontal cortex (OFC), whereas participants with depression showed greater activation than healthy subjects in the left dorsal OFC and right caudal anterior cingulate cortex. In addition, healthy adolescents, but not participants with depression, showed a negative correlation between high-risk behavior and neuronal activation in prespecified prefrontal regions. CONCLUSIONS: These results suggest subtle changes in the neural responses to reward selection in depressed adolescents. These findings should be replicated in larger samples, and the association of these neuronal changes with treatment response and prognosis should be examined. PMID- 21621148 TI - Prevalence of telomerase activity in human cancer. AB - Telomerase activity has been measured in a wide variety of cancerous and non cancerous tissue types, and the vast majority of clinical studies have shown a direct correlation between it and the presence of cancerous cells. Telomerase plays a key role in cellular immortality and tumorigenesis. Telomerase is activated in 80-90% of human carcinomas, but not in normal somatic cells, therefore, its detection holds promise as a diagnostic marker for cancer. Measurable levels of telomerase have been detected in malignant cells from various samples: tissue from gestational trophoblastic neoplasms; squamous carcinoma cells from oral rinses; lung carcinoma cells from bronchial washings; colorectal carcinoma cells from colonic luminal washings; bladder carcinoma cells from urine or bladder washings; and breast carcinoma or thyroid cancer cells from fine needle aspirations. Such clinical tests for telomerase can be useful as non invasive and cost-effective methods for early detection and monitoring of cancer. In addition, telomerase activity has been shown to correlate with poor clinical outcome in late-stage diseases such as non-small cell lung cancer, colorectal cancer, and soft tissue sarcomas. In such cases, testing for telomerase activity can be used to identify patients with a poor prognosis and to select those who might benefit from adjuvant treatment. Our review of the latest medical advances in this field reveals that telomerase holds great promise as a biomarker for early cancer detection and monitoring, and has considerable potential as the basis for developing new anticancer therapies. PMID- 21621149 TI - Increased risk of mortality from overweight and obesity in middle-aged individuals from six communities in Taiwan. AB - BACKGROUND/PURPOSE: Although obesity is becoming a major public health problem, data are limited on the relationship between body mass index (BMI) and survival in Taiwanese populations. Therefore, the aim of this study was to evaluate the long-term effects of obesity on the risk of death from any cause and from specific diseases in middle-aged Taiwanese adults. METHODS: We investigated the association between BMI and mortality in a prospective cohort study. In all, 6603 men and women (age range, 20-65 years) were included. RESULTS: The mean BMI at baseline was 22.7 kg/m2. During an average 24-year follow-up, 1896 of the 6603 individuals died (28.7%). The relationship between death from any cause and BMI followed a J-shaped pattern. Hazard ratio (HR) estimates for all-cause mortality increased among participants who were obese [HR: 1.28, 95% confidence interval (CI): 1.12-1.47 for BMI 25-26.9 kg/m2; HR: 1.46, 95% CI: 1.27-1.68 for BMI >= 27 kg/m2]. This positive association was mainly observed in deaths from diabetes, cardiovascular diseases, or cancer. The HRs for diabetes mortality were significantly higher at BMI >= 23.0 kg/m2 (HR: 1.93, 95% CI: 1.33-2.81 for BMI 23 24.9 kg/m2; HR: 2.59, 95% CI: 0.71-3.90 for BMI 25-26.9 kg/m2; and HR: 3.03, 95% CI: 2.01-4.58 for BMI >= 27 kg/m2. CONCLUSION: Increasing BMI (>= 23 kg/m2) was positively associated with deaths from diabetes and cardiovascular diseases. We found that BMI >= 25 kg/m2 was a significant predictor for all-cause mortality and >= 27 kg/m2 was a significant predictor for cancer mortality. The relationship between BMI and mortality was J-shaped in Taiwanese adults. PMID- 21621150 TI - Ischemic stroke in patients with intracranial dural arteriovenous fistulas. AB - BACKGROUND/PURPOSE: Intracranial dural arteriovenous fistulas (DAVFs) can be complicated by ischemic stroke. This study investigated the frequency and determinants of ischemic stroke in patients with intracranial DAVF. METHODS: We conducted a retrospective study of consecutive patients with intracranial DAVF. Patients with pure hemorrhagic stroke or without available brain imaging for clarifying stroke type were excluded. DAVF was diagnosed by cerebral catheter angiography. Cognard classification and location of DAVFs were ascertained. The clinical characteristics, outcome, and radiographic findings were recorded. Factors associated with occurrence of ischemic stroke in the patients with DAVFs were determined. RESULTS: A total of 134 patients were enrolled. Six patients (4.5%) had ischemic stroke (mean age: 53.8 +/- 13.4 years) and 128 patients were free from stroke (mean age: 55.4 +/- 15.2 years). Men accounted for 83% in the ischemic stroke group and 34% in the non-stroke group. Chemosis, exophthalmos and tinnitus were more frequent in the non-stroke group, whereas seizure and mental decline were more frequent in the ischemic stroke group. DAVF was associated with highest risk of ischemic stroke at locations other than the cavernous sinus or large sinuses. Ischemic stroke also correlated with types of DAVF involving cortical venous drainage, including type IIb (18%), III (15%), and IV (100%). No patient with DAVF type I and IIa had ischemic stroke. The rate of ischemic stroke in patients with concomitant DAVF and cerebral sinus thrombosis was higher than in DAVF patients without cerebral sinus thrombosis. Venous infarct was the major subtype of ischemic stroke in five DAVF patients. Endovascular therapy was the most common choice in both groups, and fewer patients in the ischemic stroke group did not receive any treatment for DAVFs. CONCLUSION: Location and type of DAVF were two important factors related to the occurrence of ischemic stroke in DAVF patients. PMID- 21621151 TI - Late diagnosis of human immunodeficiency virus infection in the era of highly active antiretroviral therapy: role of socio-behavioral factors and medical encounters. AB - BACKGROUND/PURPOSE: Despite provision of free-of-charge human immunodeficiency virus (HIV) care and antiretroviral therapy in Taiwan, a substantial proportion of patients experience late HIV diagnosis. We investigated the risk factors for late HIV diagnosis in Taiwan. METHODS: Structured interviews were conducted among newly diagnosed HIV-infected patients to collect data on demographics, socio behavioral variables and clinical profiles within 1 year preceding HIV diagnosis from August 2006 to July 2008. Multivariate analysis was performed to identify factors associated with missed opportunities for HIV testing and late HIV diagnosis (< 200 CD4 cells/MUL). RESULTS: Among 227 newly diagnosed HIV-infected patients, 107 (47%) had late HIV diagnosis. Patients who had first positive tests for HIV at voluntary testing sites [odds ratio (OR): 0.22; 95% confidence interval (CI): 0.10-0.50], regular sexual partners at HIV diagnosis (OR: 0.30; 95% CI: 0.14-0.68), and unprotected sex in the preceding 3 months (OR: 0.16; 95% CI: 0.07-0.34) were less likely to have late HIV diagnosis. Missed opportunities for HIV testing after seeking medical attention occurred in 47 patients (20.7%) and were more common in patients with late HIV diagnosis than in those who received an earlier diagnosis (23.0% vs. 15.8%, p = 0.03). Patients with late HIV diagnosis were more likely than their counterparts to have received a diagnosis of seborrheic dermatitis (7.4% vs. 0.8%, p = 0.02) and community-acquired pneumonia (5% vs. 0%, p = 0.02), for which HIV testing was not offered by the health care providers. CONCLUSION: Late HIV diagnosis is not uncommon in Taiwan. Regular risk assessment and provision of routine HIV testing in medical encounters and increase of accessibility to voluntary HIV testing could facilitate earlier diagnosis of HIV infection. PMID- 21621152 TI - Modulation of serum antinuclear antibody levels by levamisole treatment in patients with oral lichen planus. AB - BACKGROUND/PURPOSE: Serum autoantibodies, including antinuclear antibodies (ANAs), have been found in patients with oral lichen planus (OLP). This study evaluated whether Taiwanese OLP patients had significantly higher frequencies of serum ANAs than healthy control subjects, and whether levamisole treatment could modulate the antibody levels. METHODS: This study used an indirect immunofluorescence technique to measure the baseline serum levels of ANA in a group of 583 Taiwanese OLP patients and 53 healthy control subjects. Seventy-nine ANA-positive OLP patients were treated with levamisole under a regular follow-up schedule in our dental clinic, and their serum ANA levels were measured after treatment. RESULTS: We found that the frequencies of serum ANA in patients with OLP (23.2%), erosive OLP (EOLP, 23.8%), major EOLP (31.5%), and minor EOLP (18.1%) were all significantly higher than that (5.7%) in healthy control subjects. In addition, major EOLP patients had a significantly higher serum ANA positive rate than minor EOLP or non-erosive OLP patients. Of 135 ANA-positive OLP patients, 79 were treated with levamisole under a regular follow-up schedule. We found that treatment with levamisole for a period of 2-38 months (mean, 12 +/- 9 months) effectively reduced the high mean serum ANA titer (557 +/- 98) at baseline to an undetectable level (0) in all ANA-positive OLP patients, regardless of different high initial serum titers of ANA. CONCLUSION: There was a significantly higher frequency of serum ANA (23.2%) in Taiwanese OLP patients than in healthy control subjects. Treatment with levamisole for 2-38 months reduced the high serum ANA to an undetectable level, and significantly improved the signs and symptoms in all treated OLP patients. PMID- 21621153 TI - Treatment outcomes for hepatoblastoma: experience of 35 cases at a single institution. AB - BACKGROUND/PURPOSE: Hepatoblastoma is the most common malignant liver tumor in children. Comparative studies have elucidated the optimal pre- or postoperative chemotherapeutic regimens. The aim of this study was to investigate the prognostic significance of baseline tumor characteristics for overall survival and disease-free survival in children with hepatoblastoma. METHODS: There were 19 male and 16 female children with a median age of 19 months at diagnosis (range: 1 169 months) in our institution between February 1990 and June 2009. We reviewed the clinical presentation, serum alpha-fetoprotein level at diagnosis, histological subtype, treatment, and outcomes. RESULTS: Twenty-seven patients (78%) underwent neoadjuvant chemotherapy. The majority of patients subsequently underwent either hemihepatectomy (56%) or bisegmentectomy (16%). Only six patients underwent extended hepatic resection, and one of them required rescue liver transplantation. During follow-up, six patients died of progressive disease and two of perioperative mortality. Four of the six who died had pulmonary metastases at the time of diagnosis or follow-up. The median survival time was 28 months (range: 1-181 months). Five-year overall survival was 67.7% (95% confidence interval: 52.0-87.8%) and disease-free survival was 60.2% (95% confidence interval: 41.9-86.5%). CONCLUSION: The potential down-staging effect of neoadjuvant chemotherapy on hepatoblastoma might facilitate remission and convert unresectable tumors into operable ones. PMID- 21621154 TI - Effect of recombinant plasmid pEGFP-AFP-hTNF on liver cancer cells (HepG2 Cells) in vitro when delivered by PEG-PEI/Fe3O4 nanomagnetic fluid. AB - BACKGROUND/PURPOSE: Gene delivery into liver cancer cells has been a problem. This study aimed to understand the effect of using PEGI/Fe3O4 nanomagnetic fluid as a gene vector for liver cancer gene therapy. An AFP enhancer aids in the expression of tumor-specific foreign genes in AFP-producing cancer cells like HepG2 cells, and was utilized in the delivery method in this study. METHODS: We constructed recombinant plasmid PEGFP-AFP-hTNFalpha, which was transfected into AFP positive HepG2 cells and AFP negative Hela cells by PEG-PEI/Fe3O4 nanomagnetic fluid. Fluorescence microscopy was used to evaluate the transfection rate of the hTNFalpha gene in the HepG2 cells 12 hours after transfection. Reverse transcription polymerase chain reaction (RT-PCR) and western blot were used to detect expression of hTNFalpha gene in the HepG2 cells 48 hours after transfection. Methyl thiazolyl tetrazolium (MTT) assay was used to evaluate the inhibitory effect of hTNFalpha on the proliferation of HepG2 cells. Flow cytometry was used to analyze the apoptosis of HepG2 cells. RESULTS: Plasmid PEGFP-AFP-hTNFalpha delivered by PEG-PEI/Fe3O4 nanomagnetic fluid was successfully transfected into HepG2 cells and expressed in the HepG2 cells. The transfection efficacy of hTNFalpha gene delivered by PEG-PEI/Fe3O4 nanomagnetic fluid was superior to that of hTNFalpha gene delivered by lipofectamine in HepG2 cells. RT-PCR and western blot demonstrated that hTNFalpha gene was expressed in HepG2 cells that were transfected with complexes of nanomagnetic fluid/PEGFP-AFP hTNFalpha. MTT and flow cytometry showed that the hTNFalpha gene markedly exerted a cell killing effect. CONCLUSION: PEG-PEI/Fe3O4 nanomagnetic fluid successfully transfected PEGFP-AFP-hTNFalpha into HepG2 cells and induced expression of hTNFalpha gene in the HepG2 cells, thus showing promise as a gene vector for liver cancer gene therapy. Furthermore, an AFP enhancer can specifically increase the expression of target genes in cells positive for AFP. PMID- 21621155 TI - Immunogenicity and reactogenicity of diphtheria-tetanus-acellular pertussis hepatitis B-inactivated poliovirus and Haemophilus influenzae type B. AB - Immunogenicity and reactogenicity of primary vaccination with combined diphtheria tetanus-acellular pertussis-hepatitis B-inactivated poliovirus (DTPa-HBV-IPV) vaccine when co-administered with Haemophilus influenzae (Hib) conjugate vaccine were assessed in 60 healthy infants. Infants received HBV vaccine at birth, then DTPa-HBV-IPV and Hib vaccines at age 1.5 months, 3.5 months and 6 months. Blood samples were collected before the first DTPa-HBV-IPV and Hib vaccine doses and 1 month after dose 3. Reactogenicity was assessed using diary cards. One month after primary vaccination, all infants were seroprotected/seropositive against all vaccine antigens evaluated. The poliovirus antigen could not be evaluated. The vaccines were well tolerated. No case of fever > 39.0 degrees C was reported. No serious adverse events were considered related to vaccination. Primary vaccination with DTPa-HBV-IPV and Hib vaccines was immunogenic and well tolerated. Combined vaccines, such as this pentavalent vaccine, minimize the number of injections and vaccination visits required to complete primary vaccination, and provide choice and flexibility for physicians and vaccine providers. PMID- 21621156 TI - Craniofacial skeletal dysplasia of opposite-sex dizygotic twins. AB - Craniofacial skeletal dysplasia can lead to different skeletal malocclusions. Both environmental factors and heredity contribute to the formation of malocclusions. There are strong familial tendencies in the development of Angle's Class II and III malocclusions. Cases such as opposite-typed (Class II and III) malocclusions with skeletal and dentoalveolar discordance in siblings or dizygotic (DZ) twins have seldom been reported. We describe the rare case of a pair of opposite-sex DZ twins with completely different skeletal malocclusions, and discuss the clinical considerations for treatment. The patients were twins aged 13 years and 4 months. The girl had mandibular prognathism and a Class III dentoskeletal relationship, whereas the boy had skeletal Class II with mandibular retrusion. Several morphological traits have been implicated with hormonal effect. However, there was no evidence of whether the masculinization effect had any impact on jaw size in the female fetus or whether this effect lasted into adolescence. We suggest that, although DZ twins share the same growth environment, genetic or other unknown extrinsic factors can result in discordance of characteristics of the craniofacial skeleton, dentition, and occlusion. PMID- 21621157 TI - It is a tough "ACT" to follow. PMID- 21621158 TI - Pro: a cardiovascular anesthesiologist should provide services in the catheterization and electrophysiology laboratory. PMID- 21621159 TI - Con: a general anesthesiologist with a certain skill set is qualified to provide services in the interventional cardiology and electrophysiology laboratory. PMID- 21621162 TI - Antitrust risk in practice mergers: a framework for evaluation. PMID- 21621163 TI - ASMBS Position Statement on Preoperative Supervised Weight Loss Requirements. PMID- 21621164 TI - ASMBS policy statement on gastric plication. PMID- 21621165 TI - Borderline ovarian tumor and future fertility. PMID- 21621166 TI - Lamotrigine for trigeminal neuralgia: efficacy and safety in comparison with carbamazepine. AB - BACKGROUND: Anticonvulsants are regarded as useful for the treatment of neuropathic pain. In this study, we evaluated the efficacy and occurrence of side effects of lamotrigine (LTG) in comparison with carbamazepine (CBZ), in trigeminal neuralgia (TN) patients. METHODS: The study was an interventional and crossover comparison. Twenty-one patients with TN were administered with LTG in comparison to CBZ. The clinical trials comprised two phases of 40 days each, with an intervening three-day washout period. The final titration in dose for LTG was 400 mg and 1,200 mg for CBZ. Efficacy of the medications involved was determined by visual analog scale (VAS) and verbal rating scale (VRS). Side effects were recorded through marking of the profiles of side effects encountered on administration of LTG and CBZ, together with baseline haematological, hepatic and renal investigations. RESULTS: Both on VAS and VRS assessments, in terms of proportion of patients, CBZ benefitted 90.5% (19/21) of the patients with pain relief (p < 0.05), in contrast to 62% (13/21) from LTG. On VAS assessment, of the 13 patients who gained pain relief from LTG and 19 from CBZ, 77% (10/13) obtained a "complete" degree of pain relief from LTG, as compared with 21% (4/19) from CBZ. On VRS assessment, with LTG, 84% (11/13) of the patients accomplished "much better" degree of pain relief, as compared with 26% (5/19) with CBZ. On LTG, 67% (14/21) of patients endured general pharmacological side effects, as compared with 57% (12/21) of patients on CBZ (p > 0.05). Meanwhile, LTG inflicted 14% (3/21) of the patients with haematological, hepatic and renal derangements, as compared with 48% (10/21) on CBZ. CONCLUSION: LTG is generally an effective and safe treatment for management of TN, compared to CBZ. PMID- 21621167 TI - Unilateral salpingo-oophorectomy as fertility-sparing surgery for borderline ovarian tumors. AB - BACKGROUND: To investigate recurrence rates and fertility outcomes of patients with borderline ovarian tumors (BOTs) treated with fertility-sparing surgery. METHODS: This was a retrospective study. All women with BOTs from 2000 to 2006 were evaluated. Clinical outcomes were compared among groups that underwent radical, unilateral salpingo-oophorectomy, or ovarian cystectomy. The effects of clinical characteristics on recurrence were analyzed by independent t test, chi square test, and Cox proportional hazard model. RESULTS: After a mean follow-up period of 56.5 months, all 61 patients were alive. Seven (11.5%) had developed disease recurrence, and all were in the fertility-sparing group. Of these, five were in the cystectomy-only group and two in the unilateral salpingo-oophorectomy group. There was significant difference in tumor recurrence rates between the two groups (hazard ratio: 0.26, 95% confidence interval: 0.11-0.61). Nine pregnancies were achieved in six women, resulting in five deliveries CONCLUSION: Fertility sparing surgery is an acceptable and safe option for women with BOTs who wish to preserve fertility. Unilateral salpingo-oophorectomy must be considered as the first choice. PMID- 21621168 TI - Uterine arterial embolization in the management of severe post-partum hemorrhage: a successful rescue method to avoid peripartum hysterectomy. AB - BACKGROUND: Although the effects of uterine arterial embolization (UAE) for the control of post-partum hemorrhage on menstruation, fertility and future pregnancy have been reported before, domestic reports on long-term outcomes are lacking. METHODS: From April 2001 to March 2005, 9 patients who underwent UAE for post partum hemorrhage were evaluated retrospectively. The analyses included both immediate and long-term outcome, including menstruation, future fertility and subsequent pregnancies. RESULTS: The median follow-up was 82 months, ranging from 63 months to 108 months. All the women had regular menstruation after UAE treatment, but 5 had hypomenorrhea. Four women attempted pregnancy, and 3 had successful term deliveries. One woman was infertile. CONCLUSION: UAE appeared to be a safe procedure. Hypomenorrhea was common after UAE treatment (55.6%), although regular menstruation returned successfully. Future pregnancy seemed possible, and without complications. PMID- 21621169 TI - The effect of adding a home program to weekly institutional-based therapy for children with undefined developmental delay: a pilot randomized clinical trial. AB - BACKGROUND: Early rehabilitation for children with developmental delay without a defined etiology have included home and clinic programs, but no comparisons have been made and efficacy is uncertain. We compared a weekly visit for institutional based therapy (IT) to IT plus a structured home activity program (HAP). METHODS: Seventy children who were diagnosed with motor or global developmental delay (ages 6-48 months and mean developmental age 12.5 months) without defined etiology were recruited (including 45 males and 23 females). The outcomes included the comprehensive developmental inventory for infants and toddlers test and the pediatric evaluation of disability inventory. RESULTS: Children who received only IT improved in developmental level by 2.11 months compared with 3.11 months for those who received a combination of IT and HAP (p = 0.000). On all domains of the comprehensive developmental inventory for infants and toddlers test, except for self-help, children who participated in HAP showed greater improvements, including in cognition (p = 0.015), language (p = 0.010), motor (p = 0.000), and social (p = 0.038) domains. Except on the subdomain of self-care with caregiver assistance, the HAP group showed greater improvement in all the pediatric evaluation of disability inventory subdomains (p < 0.05). CONCLUSION: Early intervention programs are helpful for these children, and the addition of structured home activity programs may augment the effects on developmental progression. PMID- 21621170 TI - The quantitative detection of aripiprazole and its main metabolite by using capillary-electrophoresis. AB - BACKGROUND: We aimed to establish a feasible and reliable method to measure the level of aripiprazole and its main metabolite, dehydroaripiprazole, using a capillary-electrophoresis (CE) machine. METHODS: Two blood samples were obtained from psychiatric patients hospitalized in Yu-Li Hospital who had been treated with aripiprazole for more than 4 weeks, at least 10 mg/d. Conditions for voltage, temperature and buffer concentration was optimized on a CE machine. RESULTS: The most optimal conditions for CE were 80 mM 2-3% DMSO-phosphate as a buffer under pH = 3.0, 15 KV, 20 degrees C and a detection wavelength of 214 nm. The linear ranges of aripiprazole and dehydroaripiprazole concentration were from 0.5 to 50 ng/mL. CONCLUSION: CE method is a feasible method to measure aripiprazole level with relatively low price compared with other analytical techniques for clinical use. PMID- 21621171 TI - Effective salvage therapy of imatinib-resistant gastrointestinal stromal tumor with combination of imatinib and pegylated liposomal doxorubicin. AB - Here we presented a 60-year-old Taiwanese man with advanced gastrointestinal stromal tumor. Disease progression was noted during imatinib treatment. Surgical resection was done and mutation analysis of KIT gene in all the resected tumors revealed deletion mutations of codons 558-565 in exon 11, whereas a missense mutation was also identified at codon 822 in exon 17 in one resected tumor. Patient's disease was refractory to escalating dose of imatinib and dasatinb. Surprisingly, combination of imatinib with pegylated liposomal doxorubicin produced a substantial response and resulted in a 5-month progression free period for this imatinib-resistant gastrointestinal stromal tumor. PMID- 21621172 TI - Angioleiomyoma of the scrotal wall. AB - Angioleiomyoma of the scrotum is a rare benign lesion which can mimic a paratesticular tumor. Any solid mass within the scrotum is considered malignant until proven otherwise. Here, we present a case of an angioleiomyoma of the scrotum in a 33-year-old male who presented with painful scrotal mass. Scrotal ultrasonography demonstrated a solid mass in the scrotum, and surgical excision was carried out. Pathologic examination revealed that the tumor was angioleiomyoma. PMID- 21621173 TI - Appendiceal hemorrhage -- an uncommon cause of lower gastrointestinal bleeding. AB - Lower gastrointestinal bleeding is a common disease among elderly patients. The common sources of lower gastrointestinal bleeding include vascular disease, Crohn's disease, neoplasms, inflammatory bowel disease, hemorrhoids, and ischemic colitis. Lower gastrointestinal bleeding arising from the appendix is an extremely rare condition. We report a case of appendiceal hemorrhage in a young male. Diagnosis was made by multidetector computerized tomography during survey for hematochezia. The patient recovered well after appendectomy. The histological finding revealed focal erosion of appendix mucosa with bleeding. PMID- 21621174 TI - Pityriasis rosea following influenza (H1N1) vaccination. AB - Pityriasis rosea is a distinct papulosquamous skin eruption that has been attributed to viral reactivation, certain drug exposures or rarely, vaccination. Herein, we reported a clinicopathlogically typical case of pityriasis rosea that developed after the H1N1 vaccination. With a global H1N1 vaccination program against the pandemic H1N1 influenza, patients should be apprised of the possibility of such rare but benign skin reaction to avoid unnecessary fear. Furthermore, a brief review of the current reported skin adverse events related to the novel H1N1 vaccination in Taiwan is presented here. PMID- 21621176 TI - Research and bibliometrics: a long history.... PMID- 21621175 TI - Loss of the chromatin regulator MRG15 limits neural stem/progenitor cell proliferation via increased expression of the p21 Cdk inhibitor. AB - Chromatin regulation is crucial for many biological processes such as transcriptional regulation, DNA replication, and DNA damage repair. We have found that it is also important for neural stem/progenitor cell (NSC) function and neurogenesis. Here, we demonstrate that expression of the cyclin-dependent kinase inhibitor p21 is specifically up-regulated in Mrg15 deficient NSCs. Knockdown of p21 expression by p21 shRNA results in restoration of cell proliferation. This indicates that p21 is directly involved in the growth defects observed in Mrg15 deficient NSCs. Activated p53 accumulates in Mrg15 deficient NSCs and this most likely accounts for the up-regulation of p21 expression in the cells. We observed decreased p53 and p21 levels and a concomitant increase in the percentage of BrdU positive cells in Mrg15 null cultures following expression of p53 shRNA. DNA damage foci, as indicated by immunostaining for gammaH2AX and 53BP1, are detectable in a sub-population of Mrg15 deficient NSC cultures under normal growing conditions and the majority of p21-positive cells are also positive for 53BP1 foci. Furthermore, Mrg15 deficient NSCs exhibit severe defects in DNA damage response following ionizing radiation. Our observations highlight the importance of chromatin regulation and DNA damage response in NSC function and maintenance. PMID- 21621177 TI - Representation at different levels in a conceptual hierarchy. AB - The present study examines the influence of hierarchical level on category representation. Three computational models of representation - an exemplar model, a prototype model and an ideal representation model - were evaluated in their ability to account for the typicality gradient of categories at two hierarchical levels in the conceptual domain of clothes. The domain contains 20 subordinate categories (e.g., trousers, stockings and underwear) and an encompassing superordinate category (CLOTHES). The models were evaluated both in terms of their ability to fit the empirical data and their generalizability through marginal likelihood. The hierarchical level was found to clearly influence the type of representation: For concepts at the subordinate level, exemplar representations were supported. At the superordinate level, however, an ideal representation was overwhelmingly preferred over exemplar and prototype representations. This finding contributes to the increasingly dominant view that the human conceptual apparatus adopts both exemplar representations and more abstract representations, contradicting unitary approaches to categorization. PMID- 21621178 TI - Factors that influence search termination decisions in free recall: an examination of response type and confidence. AB - In three experiments search termination decisions were examined as a function of response type (correct vs. incorrect) and confidence. It was found that the time between the last retrieved item and the decision to terminate search (exit latency) was related to the type of response and confidence in the last item retrieved. Participants were willing to search longer when the last retrieved item was a correct item vs. an incorrect item and when the confidence was high in the last retrieved item. It was also found that the number of errors retrieved during the recall period was related to search termination decisions such that the more errors retrieved, the more likely participants were to terminate the search. Finally, it was found that knowledge of overall search set size influenced the time needed to search for items, but did not influence search termination decisions. PMID- 21621179 TI - A demonstration of direct access to colored stimuli following cueing by color. AB - To test whether cueing by color can affect orienting without first computing the location of the cued color, the impact of reorienting on the validity effect was examined. In Experiment 1 subjects were asked to detect a black dot target presented at random on either of two colored forms. The forms started being presented 750 ms before the onset of a central cue (either an arrow or a colored square). In some proportion of the trials the colors switched locations 150 ms after cue onset, simultaneously with target onset. The color switch was not found to retard responses following a color cue more than following a location cue. Furthermore, it did not reduce the validity effect of the color cue: Though the validity effect of the location cue was quite larger than the validity effect of the color cue, both effects were additive with the presence/absence of a color switch. In Experiment 2, subjects were rather asked to detect a change in shape of one of the colored forms. In this case, color switch was found to affect performance even less following a color cue. The fact that across experiments, color switch did not retard neither responding nor orienting selectively in the color cue condition, indicates that when attention is set to a certain color, reorienting to a new object following color switch does not require re-computing the address of the cued color. That finding is argued to embarrass a strong space based view of visual attention. PMID- 21621180 TI - [Blue toe syndrome as the presenting sign of a popliteal artery aneurysm]. PMID- 21621181 TI - Colloid particle and protein deposition - electrokinetic studies. AB - Recent developments in the electrokinetic determination of particle, polyelectrolyte and protein deposition at solid/electrolyte interfaces, are reviewed. In the first section basic theoretical results are discussed enabling a quantitative interpretation of the streaming current/potential and microelectrophoretic measurements. Experimental results are presented, pertinent to electrokinetic characteristics of simple (homogeneous) surfaces such as mica, silica and various polymeric surfaces used in protein studies. The influence of the ionic strength, background electrolyte composition and pH is discussed, and the effective (electrokientic) charge of these interfaces is evaluated. In the next section, experimental data obtained by streaming potential measurements for colloid particle mono- and bilayers are presented and interpreted successfully in terms of available theoretical approaches. These results, obtained for model systems of monodisperse colloid particles are used as reference data for discussion of more complicated experiments performed for polyelectrolyte and protein covered surfaces. Results are discussed, obtained for cationic polyelectrolytes (PEI, PAH) and fibrinogen adsorbing on mica, interpreted quantitatively in terms of the theoretical approach postulating a heterogeneous 3D charge distribution. The Gouy-Chapman model, based on the continuous charge distribution proved inadequate. Interesting experimental data are also discussed, obtained by electrophoretic methods in the case of protein adsorption on colloid latex particles. In the last section, supplementary results on particle deposition on heterogeneous surfaces produced by controlled protein adsorption are discussed. Quantitative relationships between the amount of adsorbed protein, zeta potential of the interface and the particle coverage are specified. Possibility of evaluating the heterogeneity of protein charge distribution is pointed out. The anomalous deposition of colloid particles on protein molecules bearing the same sign of zeta potential, which contradicts classical DLVO theory, is interpreted in terms of the fluctuation theory. It is concluded that theoretical and experimental results obtained for model colloid systems and flat interfaces can be effectively used for interpretation of protein adsorption phenomena, studied by electrophoresis. In this way the universality of electrokinetic phenomena is underlined. PMID- 21621182 TI - The antisynthetase syndrome. PMID- 21621183 TI - Clinical humanities? PMID- 21621184 TI - Clinical and inflammatory markers in amniotic fluid as predictors of adverse outcomes in preterm premature rupture of membranes. AB - OBJECTIVE: We sought to evaluate gestational age, cervical length, amniotic fluid interleukin (IL)-6, and selected proteomic biomarkers as independent predictors of adverse outcome in preterm premature rupture of membranes (PPROM). STUDY DESIGN: This was a prospective cohort study of 65 consecutive women with PPROM (20.0-34.6 weeks). Gestational age, cervical length, amniotic fluid IL-6, and proteomic biomarkers (calgranulins A and C, and neutrophil defensins 1 and 2) were evaluated at diagnosis. The predictive value for intraamniotic infection and neonatal composite morbidity was calculated by logistic regression. RESULTS: Proteomic biomarkers were independent predictors of intraamniotic infection (odds ratio, 22.1; P=.011) and neonatal composite morbidity (odds ratio, 17.6; P=.02). With the exception of a trend between gestational age and neonatal morbidity (P=.054), none of the other parameters were independent predictors of outcome measures. CONCLUSION: Selected proteomic biomarkers were the only independent predictors of adverse outcomes in PPROM. Contrary to what is reported in preterm labor with intact membranes, gestational age, cervical length, and IL-6 were not. PMID- 21621185 TI - Higher infant body fat with excessive gestational weight gain in overweight women. AB - OBJECTIVE: Gestational weight gain (GWG) is positively associated with birthweight and maternal prepregnancy body mass index (BMI) is directly related to infant fat mass (FM). This study examined whether differences exist in infant body composition based on 2009 GWG recommendations. STUDY DESIGN: Body composition was measured in 306 infants, and GWG was categorized as appropriate or excessive. Analysis of covariance was used to investigate the effects of GWG and prepregnancy BMI and their interaction on infant body composition. RESULTS: Within the appropriate group, infants from obese mothers had greater percent fat (%fat) and FM than offspring from normal and overweight mothers. Within the excessive group, infants from normal mothers had less %fat and FM than infants from overweight and obese mothers. A difference was found for %fat and FM within the overweight group between GWG categories. CONCLUSION: Excessive GWG is associated with greater infant body fat and the effect is greatest in overweight women. PMID- 21621186 TI - Effect of gestational hypercholesterolemia and maternal immunization on offspring plasma eicosanoids. AB - OBJECTIVE: Maternal immunization with oxidized low-density lipoprotein prior to pregnancy prevents pathogenic in utero programming by gestational hypercholesterolemia, but it is unknown whether gestational hypercholesterolemia and maternal immunization affect similar pathways. STUDY DESIGN: A lipidomic approach was used for unbiased plasma eicosanoid profiling in adult offspring of immunized and nonimmunized normocholesterolemic or hypercholesterolemic rabbit mothers. RESULTS: Gestational hypercholesterolemia was associated with increased levels of some eicosanoids formed by the cyclooxygenase and 12-lipoxygenase pathways only (including thromboxane B2, prostaglandin [PG] F2alpha, PGE2, and PGD2). Immunization of hypercholesterolemic or normocholesterolemic mothers reduced 9 of 14 eicosanoids of the cyclooxygenase pathway, 21 of 23 eicosanoids of the 5- and 12-lipoxygenase pathways (eg, 5-hydroxyeicosatetraenoic acid, hepoxilin B3, 12-hydroxyeicosatetraenoic acid), 8 of 19 eicosanoids of the cytochrome P-450 pathway, and all metabolites of the nonenzymatic pathway. CONCLUSION: Maternal immunization not only counteracts in utero programming by gestational hypercholesterolemia but reduces a broad range of eicosanoid modulators of immunity and inflammation in offspring. PMID- 21621187 TI - The effect of maternal obesity on the rate of failed induction of labor. AB - OBJECTIVE: The purpose of this study was to quantify the relationship between class of obesity and rate of failed induction of labor. STUDY DESIGN: Using the Ohio Department of Health's birth certificate database from January 1, 2006, through December 31, 2007, we performed a population-based cohort study that compared failed induction of labor rates between obese and normal-weight women. RESULTS: The rate of induction is associated with increasing body mass index from 28% in normal-weight women to 34% in class III obese women (body mass index, >=40 kg/m2). Induction failure rates are also associated with increasing obesity class from 13% in normal-weight women to 29% in class III obese women. Women with class III obesity without a previous vaginal delivery and a macrosomic fetus had the highest rate of failed induction at 80%. CONCLUSION: Obesity is associated with an increased risk of failed labor induction that appears to be related directly to increasing class of obesity. PMID- 21621188 TI - Characteristics of traumatic cataract wound dehiscence. AB - PURPOSE: To characterize the clinical course of cataract wound dehiscence. DESIGN: Retrospective, comparative case series. METHODS: Charts of open globe injuries (848 injuries in 846 patients) treated surgically at the Massachusetts Eye and Ear Infirmary between 2000 and 2009 were retrospectively reviewed. Time from original surgery to wound dehiscence, type of initial surgery, Ocular Trauma Score, age, gender, mechanism of injury, and visual acuity were analyzed. RESULTS: Of 846 patients with 848 open globe injuries, 63 experienced cataract wound dehiscence. The majority of these cataract wounds (89%) were extracapsular cataract extraction (ECCE), with only 7 (11%) phacoemulsification wounds. The mean patient age in the wound rupture group was 78.2 years. Female patients comprised the majority (67%) of this subpopulation. The most common mechanisms of injury were fall (65%), blunt trauma (23%), and motor vehicle accident (7%). The median raw ocular trauma score was 47 in wound dehiscence patients. Visual acuity at presentation was light perception in the wound dehiscence group. The best postoperative visual acuity was significantly worse in the wound dehiscence group (hand motion) than in the remaining patients (20/40; P=.0002). When considering the phacoemulsification patients alone, these patients fared much better, with a median postoperative vision of 20/60. CONCLUSIONS: Despite recent advances in cataract surgery, wound dehiscence remains a significant source of visual disability, mainly in the geriatric population. Rupture ECCE wound patients have a poor visual prognosis. Fortunately, patients with phacoemulsification site dehiscence appear to regain the majority of their vision after open globe repair. PMID- 21621189 TI - Levels of VEGF but not VEGF(165b) are increased in the vitreous of patients with retinal vein occlusion. AB - PURPOSE: To determine the concentration of the pro-angiogenic vascular endothelial growth factor VEGF(165) (VEGF) and the anti-angiogenic VEGF(165b) in vitreous samples of patients with branch retinal vein occlusion (BRVO) and central retinal vein occlusion (CRVO) in comparison to patients without retinal occlusive disease. DESIGN: Experimental laboratory investigation. METHODS: Vitreous samples were collected from patients undergoing surgery for arteriovenous dissection after BRVO, radial optic neurotomy after CRVO in the occlusion group, or macular pucker or macular hole in the control group. Concentrations of VEGF and VEGF(165b) were determined by ELISA and an ELISA-type antibody microarray. RESULTS: Average vitreal concentration of VEGF was 8.6 ng/mL in the CRVO group and 2.0 ng/mL in the BRVO group as compared to 0.26 ng/mL in the control group. Average vitreal concentration of VEGF(165b) was 27 pg/mL in the CRVO group, 42 pg/mL in the BRVO group, and 49 pg/mL in the control group. In patients with CRVO and BRVO, the angiogenic balance was shifted towards angiogenic stimulation. CONCLUSION: The severity of RVO from BRVO to CRVO correlates with an increase of VEGF and the decrease of VEGF(165b), indicating a pro-angiogenic shift. Altering the ratio of VEGF(165b)/VEGF(165) might be a feasible approach for treating retinal occlusive diseases. PMID- 21621190 TI - Microbicidal efficacy of thiocyanate hydrogen peroxide after adding lactoperoxidase under saliva loading in the quantitative suspension test. AB - OBJECTIVE: As shown in the quantitative suspension test adding lactoperoxidase to a thiocyanate (SCN(-)) hydrogen peroxide (H(2)O(2)) combination over the physiological saliva level has significant positive antimicrobial effects to a level of totally killing Streptococcus mutans, Streptococcus sanguinis, and Candida albicans. The aim of this study was to evaluate this positive effect under human saliva loading. METHODS: The bactericidal and fungicidal effect of lactoperoxidase was evaluated in a quantitative suspension test by using two test mixtures of a 2.0% thiocyanate and 1.2% hydrogen peroxide solution, one without (Group A) and one with (Group B) lactoperoxidase under saliva loading. Following the quantitative suspension tests (EN-13727/EN-13624), the growth of surviving bacteria and fungi in a nutrient broth was measured. The exposure times were restricted to 1, 3, 5, and 15 min. All statistical analyses were carried out with SPSS 11.5. RESULTS: In the quantitative suspension test, the combination of thiocyanate and hydrogen peroxide showed relatively low antimicrobial effectiveness on S. mutans, S. sanguinis, and C. albicans in the presence of human saliva at measured time points in comparison to the mixture with lactoperoxidase, which showed a high bactericidal activity within 15 min (S. mutans and S. sanguinis) and fungicidal activity within 3 min (C. albicans). CONCLUSION: The antimicrobial effectiveness of the tested thiocyanate hydrogen peroxide combination was increased significantly by adding lactoperoxidase in the quantitative suspension test under human saliva loading. PMID- 21621191 TI - The therapeutic potential of oxygen tension manipulation via hypoxia inducible factors and mimicking agents in guided bone regeneration. A review. AB - Intraoral bone grafting is routinely employed for implant site development prior or simultaneously to implant placement. Bone graft consolidation is a complex biological process depending on the formation of blood vessels into the augmented area. It is highly regulated by the angiogenesis and osteogenesis coupling phenomenon. The vascular system apart from supplying nutrients and oxygen to the developing and regenerating bone, also delivers critical signals which stimulate mesenchymal cell differentiation towards an osteogenic phenotype. Hypoxia inducible factors (HIFs) and mimicking agents (HMAs) (or alternatively HIF stabilizing agents) are considered to act as key stimulators of blood vessel formation. Under normoxia, HIFs are rapidly degraded. However, their degradation is prevented under hypoxia, which in turn, triggers angiogenesis. Hence, the major role of HMAs is to prevent degradation of HIFs under normoxic conditions. Recent studies suggest that HIFs and HMAs trigger the initiation and promotion of angiogenic-osteogenic cascade events. In vitro and animal studies involving genetic manipulation of individual components of the HIFs and HMAs have provided clues to how angiogenic-osteogenic coupling is achieved. Evidence from preclinical studies further suggests that topical application of HMAs enhance angiogenesis in intraoral augmented sites. In this article, we review the current understanding of the cellular and molecular mechanisms responsible for angiogenic osteogenic coupling. We also discuss the therapeutic manipulation of HIFs and HMAs in intraoral bone repair and regeneration. Such discoveries suggest promising approaches for the development of novel therapies to improve intraoral bone repair and regeneration procedures. PMID- 21621192 TI - Effect of inhaled corticosteroid on TNF-alpha production and alveolar bone loss in Wistar rats. AB - OBJECTIVE: To evaluate the effect of different concentrations of inhaled budesonide on secretion of tumoral necrosis factor-alpha (TNF-alpha) and on ligature-induced alveolar bone loss in Wistar rats. MATERIALS AND METHODS: Forty two animals were randomly divided in four groups. Control group (G1) did not receive any procedure. For the other 3 groups, alveolar bone loss was induced by placement of ligatures around the upper second molar. The contralateral molar was considered intra-group control. Group 2 (G2) was nebulized with saline solution (NaCl 0.9%). Groups 3 and 4 (G3 and G4) were nebulized with 30 MUg and 100 MUg budesonide, respectively. Administration of drugs was performed daily for 14 days. Blood samples were collected from all animals for analysis of TNF-alpha. The maxillae from G2, G3 and G4 were removed and defleshed with 9% sodium hypochlorite. Morphometric analysis of bone loss was performed in digital standard photographs. Statistical analysis was performed with one-way ANOVA, followed by Tukey HSD or Scheffe multiple comparison's test (significance level P <= 0.05). RESULTS: Mean alveolar bone loss values for teeth with ligature were 0.72, 0.70 and 0.77 mm for Groups 2, 3 and 4, respectively. No statistically significant differences were found amongst groups with or without ligature. The production of TNF-alpha was 60% higher in the presence of ligature (G1 vs. G2/G3/G4). No effect was observed in TNF-alpha secretion after inhalation of budesonide. CONCLUSION: Inhaled budesonide in different concentrations did not alter alveolar bone loss and TNF-alpha secretion in male Wistar rats. PMID- 21621193 TI - Influence of the ability to roll the tongue and tongue-training parameters on oral motor performance and learning. AB - OBJECTIVE: Training of tongue function is an important part of rehabilitation of patients with brain damage. A standardized tongue-training task has been shown to induce cortical plasticity. This study tested the possible influence of the natural ability to roll the tongue and modulations of tongue-training parameters on tongue-training performance. DESIGN: A total of 44 healthy adult subjects participated. 29 subjects (15 with and 14 without ability to roll their tongue) performed 1h standard tongue-training task. Another 15 subjects participated in 2 sessions: Standard and Modulation in randomized order. Standard session: 1h tongue-training with fixed training parameters; Modulation session: 1h tongue training with modulation of training parameters every 20 min (3 different settings - A, B, C, with different timing of task). Perceived task difficulty was evaluated on a 0-10 numerical rating scale (NRS). RESULTS: All participants improved performance during training (P<0.001). The ability to roll the tongue did not influence tongue-training performance (P=0.617). Modulation of training parameters influenced baseline training performance (P<0.018) and improvement (P=0.039). The mean perceived difficulty on NRS was: Standard: 6 +/- 2; Modulation: A: 6 +/- 2; B: 7 +/- 1;C: 4 +/- 1. Perceived task difficulty (rho= 0.740, P<0.001) and performance improvement (rho=-0.610, P<0.001) were inversely correlated with baseline training performance. CONCLUSION: The natural ability to roll the tongue did not influence tongue-training performance. Modulation of tongue-training parameters by alteration of timing of the training task influenced tongue-training performance and perceived task difficulty. PMID- 21621194 TI - Acute ether stress differentially affects corticotropin-releasing factor and urocortin 1 in the Brattleboro rat. AB - Arginine-vasopressin (AVP), corticotropin-releasing factor (CRF) and urocortin 1 (Ucn1) play a role in the stress response. The CRF-producing paraventricular nucleus of the hypothalamus (PVN), oval bed nucleus of the stria terminalis (BSTov) and central amygdala (CeA), and the Ucn1-expressing non-preganglionic Edinger-Westphal nucleus (npEW) all possess AVP receptors. We hypothesized that AVP is involved in the response of these four brain centers to acute physiological (ether) stress. To test this hypothesis, we studied AVP-deficient Brattleboro (BB) rats using quantitative immunocytochemistry. First, we showed that non-stressed wild-type (WT) and BB rats did not differ from each other in Fos contents, indicating similar (immediate early) gene expression activity, but that in BB rats CRF contents were lower in the PVN and higher in the CeA. Second, we found that stress induced Fos response in the PVN, CeA and npEW with strengths different for each center, but similar for BB and WT rats. Finally, no effects of stress on CRF and Ucn1 contents were seen in the WT rat brain, but in BB rats stress increased CRF contents in the PVN, and the CeA revealed more CRF in stressed BB than in WT rats. On the basis of these results we propose that during acute stress AVP interacts with, especially, the PVN and the CeA, to change their rates of biosynthesis and/or release of CRF. PMID- 21621195 TI - Contextual influences on rapid object categorization in natural scenes. AB - The current study aimed to investigate the effects of scene context on rapid object recognition using both behavioral and electrophysiological measures. Participants performed an animal/non-animal go/no-go categorization task in which they had to decide whether or not a flashed scene contained an animal. Moreover, the influence of scene context was manipulated either by retaining, deleting, or phase-randomizing the original scene background. The results of Experiments 1 and 2 showed that participants responded more accurately and quickly to objects appearing with their original scene backgrounds. Moreover, the event-related potential (ERP) data obtained from Experiment 2 showed that the onset latency of the frontal go/no-go ERP difference was delayed for objects appearing with phase randomized scene backgrounds compared to objects appearing with their original scene backgrounds, providing direct evidence that scene context facilitates object recognition. Additionally, an increased frontal negativity along with a decreased late positive potential for processing objects presented in meaningless scene backgrounds suggest that the categorization task becomes more demanding when scene context is eliminated. Together, the results of the current study are consistent with previous research showing that scene context modulates object processing. PMID- 21621196 TI - Chronic unpredictable stress induces a reversible change of PER2 rhythm in the suprachiasmatic nucleus. AB - Many clinical studies have shown that circadian rhythm abnormalities are strongly associated with major depression. The master clock of the circadian system in mammals is located in the suprachiasmatic nucleus (SCN) within the anterior hypothalamus, where Per1 and Per2 are essential core components of circadian rhythm oscillation. Chronic unpredictable stress (CUS) is a reliable animal model of depression with good face, predictive, and constructive validity. In the present study, we investigated the effects of CUS on the circadian expression of PER1 and PER2 in the SCN. We found that CUS led to depressive-like behavior and reduced the amplitude of PER2 oscillation in the SCN, which were blocked by 3 weeks of desipramine (DMI) treatment. 2 weeks after termination of CUS, the decreased peak of PER2 expression returned to control levels, whereas depressive like behavior remained unchanged. Our findings suggest that the dampened amplitude of PER2 expression in the SCN may participate in the development of depressive-like behavior induced by CUS but is unlikely involved in the long lasting effects of CUS on depressive-like behavior. PMID- 21621197 TI - Tunable stereoselectivity during sialylation using an N-acetyl-5-N,4-O oxazolidinone-protected p-toluene 2-thio-sialoside donor with Tf(2)O/Ph(2)SO/TTBPy. AB - An N-acetyl-5-N,4-O-oxazolidinone-protected p-toluene 2-thio-sialoside donor, promoted by Ph(2)SO/Tf(2)O/TTBPy, was thoroughly investigated in the coupling to various acceptors. The stereoselectivity of the sialylation was found to be dependent on the various reaction conditions, such as pre-activation time, reaction time, the amount of Ph(2)SO, and TTBPy. A detailed Ph(2)SO/Tf(2)O promoted glycosylation mechanism was proposed, which contained three crucial reactive species: an oxacarbenium ion, C2-sialyloxosulfonium salts, and oxosulfonium supramers. Our research results indicate that it is possible to tune the stereoselectivity of the sialylation by carefully changing the reaction conditions. For instance, Ph(2)SO (2.0-3.0 equiv)/TTBPy (0-1.0 equiv) promotion gives higher alpha-selective sialylation in dichloromethane, while Ph(2)SO (4-5 equiv)/TTBPy (0 equiv) or Ph(2)SO (2.0 equiv)/TTBPy (2.0 equiv) affords lower stereoselectivity. PMID- 21621198 TI - Expression of the inflammatory chemokines CCL2, CCL5 and CXCL2 and the receptors CCR1-3 and CXCR2 in T lymphocytes from mammary tumor-bearing mice. AB - Chemokines and their receptors have been studied in several solid tumor models as mediators of inflammation. In turn, inflammation has been implicated in the promotion and progression of tumors, and as such, chemokines have been proposed as novel molecular targets for chemotherapy. While the expression of these molecules has been described in tumor cells, endothelial cells, macrophages and neutrophils, less attention has been paid to the expression profile of these molecules by T lymphocytes in the periphery or infiltrating the tumor. Using the D1-DMBA-3 murine mammary adenocarcinoma model, we aimed to better characterize the differential expression of chemokines and/or their receptors in the host and in the tumor microenvironment, and specifically, in the T cells of tumor-bearing mice compared to normal control animals. We found that T lymphocytes from tumor bearing mice express the pro-inflammatory chemokines, CCL2, CCL5 and CXCL2, as well as the chemokine receptors, CCR1, CCR2, CCR3 and CXCR2. PMID- 21621199 TI - Diffusion-weighted MRI of the transplanted liver. AB - AIM: To assess the value of diffusion-weighted imaging (DWI) in evaluating parenchymal disorders following orthotopic liver transplantation (OLT). MATERIALS AND METHODS: This institutional review board-approved, retrospective study measured the hepatic apparent diffusion coefficients (ADC) in patients following OLT. Those with vascular complications or within 3 months of OLT were excluded. A single-shot echoplanar sequence with b values of 50, 400 (or 500), and 800 s/mm(2) was performed. Liver biopsy specimens [performed with a median of 17 days after magnetic resonance imaging (MRI)] were recorded for the presence and severity of parenchymal disorders, such as acute cellular rejection, and recurrence of fibrosis in all patients, and the recurrence of viral hepatitis in patients with hepatitis C. ADC values were measured blinded to histology in 41 patients (33 males) who had 56 MRI scans. RESULTS: There was a significant difference in ADC values associated with a histological abnormality seen on core biopsy [n=43, mean (SD) ADC of 0.91 (0.15)*10(-3) mm(2)/s] and those associated with no histological abnormality [n=13, mean (SD) ADC of 1.11 (0.17)*10(-3) mm(2)/s; (p=0.003)]. ADC values did not predict any of the individual parenchymal disorders on logistic regression analysis. When the ADC value was <0.99*10(-3) mm(2)/s, there was a sensitivity and specificity of 85% and 72%, respectively, in predicting a parenchymal disorder (area under ROC curve=0.84; 95% CI 0.72 to 0.92). CONCLUSION: ADC measurements may help in deciding which patients require core liver biopsy after OLT. However, ADC values are not likely to be reliable in differentiating between the various parenchymal disorders. PMID- 21621200 TI - DFAspike: a new computational proposition for efficient recognition of epileptic spike in EEG. AB - An automated method has been presented for the detection of epileptic spikes in the electroencephalogram (EEG) using a deterministic finite automata (DFA) and has been named as DFAspike. EEG data (sampled, 256 Hz) files are the inputs to the DFAspike. The DFAspike was tested with different data files containing epileptic spikes. The obtained recognition rate of epileptic spike was 99.13% on an average. This system does not require any kind of prior training or human intrusion. The result shows that the designed system can be very effectively used for the detection of spikes present in the recorded EEG signals. PMID- 21621201 TI - Fetal development assessed by heart rate patterns--time scales of complex autonomic control. AB - The increasing functional integrity of the organism during fetal maturation is connected with increasing complex internal coordination. We hypothesize that time scales of complexity and dynamics of heart rate patterns reflect the increasing inter-dependencies within the fetal organism during its prenatal development. We investigated multi-scale complexity, time irreversibility and fractal scaling from 73 fetal magnetocardiographic 30min recordings over the third trimester. We found different scale dependent complexity changes, increasing medium scale time irreversibility, and increasing long scale fractal correlations (all changes p<0.05). The results confirm the importance of time scales to be considered in fetal heart rate based developmental indices. PMID- 21621202 TI - Stochastic cellular automata model and Monte Carlo simulations of CD4+ T cell dynamics with a proposed alternative leukapheresis treatment for HIV/AIDS. AB - Acquired Immunodeficiency Syndrome (AIDS) is responsible for millions of deaths worldwide. To date, many drug treatment regimens have been applied to AIDS patients but none has resulted in a successful cure. This is mainly due to the fact that free HIV particles are frequently in mutation, and infected CD4(+) T cells normally reside in the lymphoid tissue where they cannot (so far) be eradicated. We present a stochastic cellular automaton (CA) model to computationally study what could be an alternative treatment, namely Leukapheresis (LCAP), to remove HIV infected leukocytes in the lymphoid tissue. We base our investigations on Monte Carlo computer simulations. Our major objective is to investigate how the number of infected CD4(+) T cells changes in response to LCAP during the short-time (weeks) and long-time (years) scales of HIV/AIDS progression in an infected individual. To achieve our goal, we analyze the time evolution of the CD4(+) T cell population in the lymphoid tissue (i.e., the lymph node) for HIV dynamics in treatment situations with various starting times and frequencies and under a no treatment condition. Our findings suggest that the effectiveness of the treatment depends mainly on the treatment starting time and the frequency of the LCAP. Other factors (e.g., the removal proportion, the treatment duration, and the state of removed cells) that likely influence disease progression are subjects for further investigation. PMID- 21621203 TI - Society and the reception of imaging technology: the American experience. PMID- 21621204 TI - Complex visual hallucinations after occipital extrastriate ischemic stroke. PMID- 21621205 TI - Xenotransplantation of cryopreserved ovarian tissue from patients with ovarian tumors into SCID mice--no evidence of malignant cell contamination. AB - This study examined the possible presence of malignant cells in ovarian cortex from patients with ovarian tumors after xenografting of the ovarian tissue into severe combined immunodeficiency mice. None of the mice presented symptoms of reintroduced malignancy nor did microscopic and immunohistochemical evaluation of the grafts raise any suspicion of residual malignant disease. PMID- 21621206 TI - Ultrastructure of tubular smooth endoplasmic reticulum aggregates in human metaphase II oocytes and clinical implications. AB - OBJECTIVE: To compare demographic, embryologic, pregnancy, and newborn outcomes after intracytoplasmic sperm injection (ICSI) cycles with or without mature oocytes (metaphase II [MII]) showing visible aggregates of tubular smooth endoplasmic reticulum (aSERT) and to describe the ultrastructure of this dysmorphism. DESIGN: Retrospective study. SETTING: Private fertility center and university cell biology and genetics departments. PATIENT(S): There were 721 ICSI cycles, 520 carrying morphologically normal MII (control group) and 60 containing aSERT-MII (study group). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Embryologic and clinical and live birth outcomes, including malformations and ultrastructural characterization of aSERT-MII. RESULT(S): Compared with the control group there was a significant decrease in the fertilization, embryo cleavage, and blastocyst rates in the study group. The only child born after transfer of embryos derived from aSERT-MII presented a major cardiovascular malformation. Ultrastructurally, large aSERT were surrounded by abnormal-shaped mitochondria and clusters of small dense bodies formed by very small vesicles, and they had curvilinear dense tubules in the interior. The same pathology was observed in small peripheral aSERT. CONCLUSION(S): The presence of large aSERT, showing attainment of the periphery, demonstrated that the cytoplasm is pathologic. The compromised embryo development and implantation was associated with decreased clinical outcomes and newborn malformations. Therefore, oocytes with large aSERT should not be used for embryo transfer. PMID- 21621207 TI - In smokers, swim-up and discontinuous gradient centrifugation recover spermatozoa with equally lower amounts of DNA damage than spermatozoa obtained from neat semen. AB - We compared the abilities of two commonly used semen preparation techniques to decrease the amount of benzo(a)pyrene-diol-epoxide (BPDE)-DNA adducts in the spermatozoa of ten smokers. Semen processing by swim-up or discontinuous gradient centrifugation recovered spermatozoa showing an equally significantly lower amount of BPDE-DNA adducts than in unprepared spermatozoa from neat semen. PMID- 21621208 TI - Handling packet dropouts and random delays for unstable delayed processes in NCS by optimal tuning of PIlambdaDMU controllers with evolutionary algorithms. AB - The issues of stochastically varying network delays and packet dropouts in Networked Control System (NCS) applications have been simultaneously addressed by time domain optimal tuning of fractional order (FO) PID controllers. Different variants of evolutionary algorithms are used for the tuning process and their performances are compared. Also the effectiveness of the fractional order PI(lambda)D(MU) controllers over their integer order counterparts is looked into. Two standard test bench plants with time delay and unstable poles which are encountered in process control applications are tuned with the proposed method to establish the validity of the tuning methodology. The proposed tuning methodology is independent of the specific choice of plant and is also applicable for less complicated systems. Thus it is useful in a wide variety of scenarios. The paper also shows the superiority of FOPID controllers over their conventional PID counterparts for NCS applications. PMID- 21621209 TI - Severe brachial plexus injuries in rugby. AB - We describe the mechanisms, pattern of injuries, management and outcomes of severe injuries to the brachial plexus sustained during the play of rugby. Thirteen cases of severe injury to the brachial plexus caused by tackles in rugby had detailed clinical assessment, and operative exploration of the brachial plexus. Seventeen spinal nerves were avulsed, two were ruptured and there were traction lesions in continuity of 24 spinal nerves. The pattern of nerve lesion was related to the posture of the neck and the forequarter at the moment of impact. Early repair by nerve transfer enabled some functional recovery, and decompression of lesions in continuity was followed by recovery of nerve function and relief of pain. PMID- 21621210 TI - Circulating levels of bisphenol A and phthalates are related to carotid atherosclerosis in the elderly. AB - BACKGROUND AND OBJECTIVE: Bisphenol A (BPA) levels have previously been associated with coronary heart disease (CHD). Since CHD is an atherosclerotic disease, we investigated if circulating levels of BPA and phthalate metabolites are related to atherosclerosis in a cross-sectional study. METHODS: In the population-based Prospective Investigation of the Vasculature in Uppsala Seniors (PIVUS) study (1016 subjects all aged 70), the prevalence of overt plaques and echogenectity (grey scale median, GSM) of carotid artery plaques were recorded by ultrasound in both of the carotid arteries. The thickness (IMT) and echogenicity (IM-GSM) of the intima-media complex were also measured. Bisphenol A (BPA) and 10 phthalate metabolites were analyzed in serum by a API 4000 liquid chromatograph/tandem mass spectrometer. RESULTS: Mono-methyl phthalate (MMP) was related to carotid plaques in an inverted U-shaped manner. This pattern was significant after adjustment for gender, body mass index, blood glucose, blood pressure, HDL and LDL-cholesterol, serum triglycerides, smoking, antihypertensive treatment and statin use (p=0.004). High levels of BPA, mono-isobutyl phthalate (MiBP) and MMP were associated with an echogenic IM-GSM and plaque GSM, while high levels of mono-2-ethylhexyl phthalate (MEHP) were associated with an echolucent IM-GSM and plaque GSM (p<0.0001 after adjustment). CONCLUSION: The phthalate metabolite MMP was related to atherosclerotic plaques in an inverted U shaped manner independently of CV risk factors. Some phthalates and BPA were also related to the echogenicity of the plaques, suggesting a role for plaque associated chemicals in atherosclerosis. PMID- 21621211 TI - Knee and ankle joint torque-angle relationships of multi-joint leg extension. AB - The force-length-relation (F-l-r) is an important property of skeletal muscle to characterise its function, whereas for in vivo human muscles, torque-angle relationships (T-a-r) represent the maximum muscular capacity as a function of joint angle. However, since in vivo force/torque-length data is only available for rotational single-joint movements the purpose of the present study was to identify torque-angle-relationships for multi-joint leg extension. Therefore, inverse dynamics served for calculation of ankle and knee joint torques of 18 male subjects when performing maximum voluntary isometric contractions in a seated leg press. Measurements in increments of 10 degrees knee angle from 30 degrees to 100 degrees knee flexion resulted in eight discrete angle configurations of hip, knee and ankle joints. For the knee joint we found an ascending-descending T-a-r with a maximum torque of 289.5 degrees +/- 43.3 Nm, which closely matches literature data from rotational knee extension. In comparison to literature we observed a shift of optimum knee angle towards knee extension. In contrast, the T-a-r of the ankle joint vastly differed from relationships obtained for isolated plantar flexion. For the ankle T-a-r derived from multi-joint leg extension subjects operated over different sections of the force-length curve, but the ankle T-a-r derived from isolated joint efforts was over the ascending limb for all subjects. Moreover, mean maximum torque of 234.7 +/- 56.6 Nm exceeded maximal strength of isolated plantar flexion (185.7 +/- 27.8 Nm). From these findings we conclude that muscle function between isolated and more physiological multi-joint tasks differs. This should be considered for ergonomic and sports optimisation as well as for modelling and simulation of human movement. PMID- 21621212 TI - Fast and sensitive high performance liquid chromatography analysis of cosmetic creams for hydroquinone, phenol and six preservatives. AB - A fast and sensitive HPLC method for analysis of cosmetic creams for hydroquinone, phenol and six preservatives has been developed. The influence of sample preparation conditions and the composition of the mobile phase and elution mode were investigated to optimize the separation of the eight studied components. Final conditions were 60% methanol and 40% water (v/v) extraction of the cosmetic creams. A C18 column (100 mm * 2.1 mm) was used as the separation column and the mobile phase consisted of methanol and 0.05 mol/L ammonium formate in water (pH=3.0) with gradient elution. The results showed that complete separation of the eight studied components was achieved within 10 min, the linear ranges were 1.0-200 MUg/mL for phenol, 0.1-150 MUg/mL for sorbic acid, 2.0-200 MUg/mL for benzoic acid, 0.5-200 MUg/mL for hydroquinone, methyl paraben, ethyl paraben and propyl paraben, butyl paraben, and good linear correlation coefficient (>=0.9997) were obtained, the detection limit was in the range of 0.05-1.0 MUg/mL, the average recovery was between 86.5% and 116.3%, and the relative standard deviation (RSD) was less than 5.0% (n=6). The method is easy, fast and sensitive, it can be employed to analyze component residues in cosmetic creams especially in a quality control setting. PMID- 21621213 TI - Simple, high throughput ultra-high performance liquid chromatography/tandem mass spectrometry trace analysis of perfluorinated alkylated substances in food of animal origin: milk and fish. AB - The present study documents development and validation of a novel approach for determination of 23 perfluorinated alkylated substances (PFASs) in food of animal origin represented by milk and fish. The list of target analytes comprises four classes of PFASs, both ionic and non-ionic: 11 perfluorocarboxylic acids (PFCAs), 4 perfluorosulphonic acids (PFSAs), 5 perfluorosulphonamides (FOSAs) and 3 perfluorophosphonic acids (PFPAs). Fast sample preparation procedure is based on an extraction of target analytes with acetonitrile (MeCN) and their transfer (supported by inorganic salts and acidification) into the organic phase. Removing of matrix co-extracts by a simple dispersive solid phase extraction (SPE) employing ENVI-Carb and C18 sorbents is followed by an efficient sample pre concentration performed by acetonitrile evaporation and subsequent dilution of residue in a small volume of methanol (matrix equivalent in the final extracts was 16 and 8 g mL(-1), for milk and fish respectively). Using modern instrumentation consisting of ultra-high performance liquid chromatography (UHPLC) hyphenated with a tandem mass spectrometer (MS/MS), limits of quantification (LOQs) as low as 0.001-0.006 MUg kg(-1) for milk and 0.002-0.013 MUg kg(-1) for fish can be achieved. Under these conditions, a wide spectrum of PFASs, including minor representatives, can be determined which enables collecting data required for human exposure studies. The pilot study employing the new method for examination of milk and canned fish samples was realized. Whereas in majority of canned fish products a wide spectrum of PFCAs, perfluorooctanesulphonic acid (PFOS) and perfluoro-1-octanesulphonamide (PFOSA) was detected, only in a few milk samples very low concentrations (LOQ levels) of PFOS and perfluorooctansulphonic acid (PFDS) were found. PMID- 21621214 TI - Flow field-flow fractionation for the analysis and characterization of natural colloids and manufactured nanoparticles in environmental systems: a critical review. AB - The use of flow field flow fractionation (FlFFF) for the separation and characterization of natural colloids and nanoparticles has increased in the last few decades. More recently, it has become a popular method for the characterization of manufactured nanoparticles. Unlike conventional filtration methods, FlFFF provides a continuous and high-resolution separation of nanoparticles as a function of their diffusion coefficient, hence the interest for use in determining particle size distribution. Moreover, when coupled to other detectors such as inductively coupled plasma-mass spectroscopy, light scattering, UV-absorbance, fluorescence, transmission electron microscopy, and atomic force microscopy, FlFFF provides a wealth of information on particle properties including, size, shape, structural parameters, chemical composition and particle-contaminant association. This paper will critically review the application of FlFFF for the characterization of natural colloids and natural and manufactured nanoparticles. Emphasis will be given to the detection systems that can be used to characterize the nanoparticles eluted from the FlFFF system, the obtained information and advantages and limitation of FlFFF compared to other fractionation and particle sizing techniques. This review will help users understand (i) the theoretical principles and experimental consideration of the FlFFF, (ii) the range of analytical tools that can be used to further characterize the nanoparticles after fractionation by FlFFF, (iii) how FlFFF results are compared to other analytical techniques and (iv) the range of applications of FlFFF for natural and manufactured NPs. PMID- 21621215 TI - Electrokinetic extraction on artificial liquid membranes of amphetamine-type stimulants from urine samples followed by high performance liquid chromatography analysis. AB - Electromembrane extraction (EME) coupled with high performance liquid chromatography and ultraviolet detection was developed for determination of amphetamine-type stimulants in human urine samples. Amphetamines migrated from 3 mL of different human urine matrices, through a thin layer of 2-nitrophenyl octyl ether (NPOE) containing 15% tris-(2-ethylhexyl) phosphate (TEHP) immobilized in the pores of a porous hollow fiber, and into a 15 MUL acidic aqueous acceptor solution present inside the lumen of the fiber. Equilibrium extraction conditions were obtained after 7 min of operation. Experimental design and response surface methodology (RSM) were used for optimization of EME parameters. Under optimal conditions, amphetamines were effectively extracted with recoveries in the range of 54-70%, which corresponded to preconcentration factors in the range of 108 140. The calibration curves were investigated in the range of 0-7 MUg mL(-1) and good linearity was achieved with a coefficient of estimation better than 0.991. Detection limits and inter-day precision (n=3) were less than 0.01 MUg mL(-1) and 11.2%, respectively. PMID- 21621216 TI - Synthesis and foaming properties of new anionic surfactants based on a renewable building block: sodium dodecyl isosorbide sulfates. AB - Two agro-based anionic surfactants containing an isosorbide moiety have been synthesized and their amphiphilic properties evaluated. Since isosorbide is now considered as an important platform chemical of the starch industry, these compounds could represent bio-sourced alternatives to the alkyl ether sulfates (notably lauryl ether sulfate, LES) that are based on petroleum-derived ethylene oxides. As isosorbide is an asymmetric diol, two isomers can be prepared (2-O dodecyl isosorbide sulfate and 5-O-dodecyl isosorbide sulfate) that show significantly different aqueous properties as regards to their Krafft temperatures and critical micellar concentrations. 5-O-dodecyl isosorbide sulfate is the most soluble and the most efficient surfactant. It possesses a much lower critical micelle concentration (cmc) than sodium dodecyl sulfate, SDS, leading to comparable foaming properties with a three times lower concentration. Its behavior compares well with the one of pure diethoxylated dodecyl sulfate that has also been prepared and evaluated in this work. PMID- 21621217 TI - Controllable synthesis and luminescent properties of three-dimensional nanostructured CaWO4:Tb3+ microspheres. AB - Three-dimensional (3D) nanostructured CaWO(4):Tb(3+)microspheres assembled by submicrospindles were synthesized via a mild sonochemical route from an aqueous solution of CaCl(2), TbCl(3) and Na(2)WO(4) with the aid of surfactant Polyglycol 600 (PEG-600). The crystal structure and morphology of the as-prepared products were characterized by using X-ray diffraction (XRD) and field emission scanning electron microscopy (FE-SEM). Rietveld refinement was carried out on the XRD data. The results showed that the CaWO(4):Tb(3+)nanoparticles can be formed without ultrasonic irradiation or addition of PEG-600. With continuously increasing irradiation time the submicrospindles and microspheres could be self assembled. The central diameter and length of the submicrospindles are around 190 and 500 nm, respectively. The 3D CaWO(4):Tb(3+)nanostructured microspheres with diameter of 2-4 MUm were assembled by the submicrospindles. A possible formation mechanism for the 3D-structured CaWO(4):Tb(3+)microspheres was proposed. The Photoluminescent (PL) properties of Tb(3+) ions in the nanostructured CaWO(4) microspheres were studied. The energy transfer processes in CaWO(4):Tb(3+)microspheres were analyzed. The electric dipole-dipole energy transfers related to (5)D(3) level were studied by inspecting the fluorescence decay of (5)D(3) level. The energy transfer critical distance was estimated. PMID- 21621218 TI - Fourier transform rheology as a universal non-linear mechanical characterization of droplet size and interfacial tension of dilute monodisperse emulsions. AB - A new protocol to gain interfacial tension and droplet size of dilute monodisperse emulsions from Fourier Transform Rheology (FTR), is proposed. Specifically, a universal dimensionless quantity E was found at small strain amplitudes to correlate with the droplet size of the emulsion where E is inversely related to the square of the capillary number Ca and directly proportional to the relative intensities of the fifth and third harmonics, I(5)/I(3). The limiting value E(0) at small strain deformations can be used as a universal parameter to calculate different emulsion properties. Different morphological constitutive models for emulsions were used to establish the universality of the parameter E(0). Preliminary analysis on experimental data confirms the validity of this approach for the characterization of emulsion properties, including the estimation of interfacial tension and droplet radius. PMID- 21621219 TI - Searching for factors underlying cerebral plasticity in the normal and injured brain. AB - Brain plasticity refers to the capacity of the nervous system to change its structure and ultimately its function over a lifetime. There have been major advances in our understanding of the principles of brain plasticity and behavior in laboratory animals and humans. Over the past decade there have been advances in the application of these principles to brain-injured laboratory animals. To date, there have been few major applications of this knowledge to establish postinjury interventions in humans. A significant challenge for the next 20 years will be the translation of this work to improve the outcome from brain injury and disease in humans. The goal of this review is to synthesize the multidisciplinary laboratory work on brain plasticity and behavior in the injured brain to inform the development of rehabilitation programs. LEARNING OUTCOMES: Readers will be able to: (a) identify principles of brain plasticity, (b) review the application of these principles to the treatment of brain-injured laboratory animals, and (c) consider the translation of the new treatments to brain-injured humans. PMID- 21621220 TI - Children with oligoarticular juvenile idiopathic arthritis are at considerable risk for growth retardation. AB - OBJECTIVE: To assess linear growth in patients with persistent oligoarticular juvenile idiopathic arthritis (JIA) treated by intra-articular corticosteroid injections (IACSI). STUDY DESIGN: Data were obtained from a retrospective review of the charts of 95 patients with persistent oligoarticular JIA (69 females). The mean age at first visit was 4.9 +/- 3.4 years, with follow-up of 6 +/- 3.7 years. The height SDS for chronologic age (z-score) was correlated with the clinical course of the disease and compared among patients treated by IACSI alone (group I) or by a combination of disease-modifying antirheumatic drugs (DMARDs) (group II). RESULTS: Growth retardation was found in 35.8% of patients (Delta z-score < 0.3), including 11.6% with severe growth retardation (Delta z-score <-1.0). Growth retardation was found in a smaller proportion of patients in group I (any growth retardation, 30.6%; severe growth retardation, 6.5%) than in patients in group II (any growth retardation, 44.4%; severe growth retardation, 21.2%; P < .05). Elevated erythrocyte sedimentation rate values (>= 40 mm/1sth) indicated a significantly higher risk for growth retardation. All other clinical variables had no association with growth retardation. CONCLUSION: A significant proportion of patients with persistent oligoarticular JIA have growth retardation and a minority have severe growth retardation. Only elevated erythrocyte sedimentation rate values were proven to be a good predictor of risk for growth retardation. PMID- 21621221 TI - Characterization and outcomes of young infants with acute liver failure. AB - OBJECTIVE: To characterize infants aged <= 90 days enrolled in an international, multicenter, prospective registry of children aged < 18 years with acute liver failure (ALF). STUDY DESIGN: The Pediatric Acute Liver Failure (PALF) Study Group collects prospective data on children from birth to 18 years. We analyzed data from infants aged <= 90 days enrolled in the PALF Study before May 18, 2009. RESULTS: A total of 148 infants were identified in the PALF registry (median age, 18 days). Common etiologies of ALF were indeterminate (38%), neonatal hemochromatosis (13.6%), and herpes simplex virus (12.8%). Spontaneous survival occurred in 60% of the infants, 16% underwent liver transplantation, and 24% died without undergoing liver trsansplantation. Infants with indeterminate ALF were more likely to undergo liver transplantation than those with viral-induced ALF (P = .0002). The cumulative incidence of death without liver transplantation was higher in infants with viral ALF (64%) compared with those with neonatal hemochromatosis (16%) or indeterminate ALF (14%) (P = .0007). CONCLUSION: ALF in young infants presents unique diagnostic considerations. Spontaneous survival is better than previously thought. Liver transplantation provides an additional option for care. PMID- 21621222 TI - Quality of life of adolescent kidney transplant recipients. AB - OBJECTIVE: To elicit utility-based quality of life (QOL) of adolescent kidney transplant recipients. STUDY DESIGN: We measured QOL in adolescent transplant recipients by using a visual analog scale (VAS), and 2 utility-based QOL measures, the Health Utilities Index (HUI) Mark 2/3 (HUI2/3), and the time trade off. Participants aged 11-19 years old were recruited from 5 transplantation centers in Australia. Mean scores were compared by using paired t tests, and linear multiple regression was used to define predictors for time trade-off QOL weights. RESULTS: Twenty-six adolescents participated in the study. On a scale with extremes of 0 (death) and 1 (full health), the participants had a mean (SD) time trade-off QOL weight of 0.99 +/- 0.01 and HUI2/3 utility scores of 0.86 +/- 0.16 and 0.85 +/- 0.21, respectively. Time trade-off values were significantly higher than HUI Mark 2 values (P = .01) and HUI Mark 3 values (P = .02). From the HUI measure, decrements were observed in specific QOL domains, including vision, emotion (depression and anxiety), cognition, and pain. CONCLUSION: Adolescent kidney transplant recipients had consistent and high values for their current QOL, which suggests that they perceive themselves to be close to full health. However, adequate emotional and cognitive support may improve their overall QOL. PMID- 21621223 TI - Plexiform neurofibromas in children with neurofibromatosis type 1: frequency and associated clinical deficits. AB - OBJECTIVES: To define the frequency and clinical features of plexiform neurofibromas (PN) in children with neurofibromatosis type 1. STUDY DESIGN: Sixty five children received whole-body magnetic resonance imaging (MRI) and clinical neurologic examination. Tumor sizes were calculated volumetrically with the program MedX v3.42. chi(2) test, Fisher exact test, t test, and Spearman rank correlation were used for statistical analysis. RESULTS: Seventy-three tumors were detected in 37 of these 65 children. The mean volume of the tumors was 145.4 mL or 4.8 mL/kg body weight. Eighteen of the 73 PNs caused clinical deficits in 17 children, and the other 56 PNs in 20 children were asymptomatic. Symptomatic tumors were larger than asymptomatic ones (9.6 vs 3.3 mL/kg body weight; P = .01). However, in certain body regions, for example, the head, small tumors also caused clinical deficits. Ten of 18 children >=11.5 years (median age of the 37 children with PNs) had symptomatic PNs compared with 7 of 19 who were <11.5 years (P = .25). CONCLUSION: PNs cause clinical deficits in young children. Early detection and regular MRI monitoring help to estimate growth and possible upcoming complications, and are thus beneficial for optimizing treatment and management. PMID- 21621224 TI - Corpus callosum atrophy as a predictor of age-related cognitive and motor impairment: a 3-year follow-up of the LADIS study cohort. AB - The aim of this 3-year follow-up study was to investigate whether corpus callosum (CC) atrophy may predict future motor and cognitive impairment in an elderly population. On baseline MRI from 563 subjects with age-related white matter changes (ARWMC) from the Leukoaraiosis And DISability (LADIS) study, the CC was segmented and subdivided into five anterior-posterior regions (CC1-CC5). Associations between the CC areas and decline in motor performance and cognitive functions over a 3-year period were analyzed. CC atrophy at baseline was significantly associated with impaired cognitive performance (p<0.01 for CC1, p<0.05 for CC5), motor function (p<0.05 for CC2 and CC5), and walking speed (p<0.01 for CC2 and CC5, p<0.05 for CC3 and total CC), and with development of dementia at 3 years (p<0.05 for CC1) after correction for appropriate confounders (ARWMC volume, atrophy, age, gender and handedness). In conclusion, CC atrophy, an indicator of reduced functional connectivity between cortical areas, seems to contribute, independently of ARWMC load, to future cognitive and motor decline in the elderly. PMID- 21621225 TI - Premotor cognitive status in a cohort of incident Parkinson disease patients (NEDICES). AB - BACKGROUND: A variety of symptoms may precede the classical motor features of Parkinson disease (PD). However, it is not known whether cognitive dysfunction precedes the motor phase of PD. We examined whether patients with incident PD had had global cognitive function disturbances three years prior to diagnosis when compared with matched controls in a cohort of community-dwelling subjects. METHODS: All participants were age 65 years or older (median 76 years) and were enrolled in the Neurological Disorders in Central Spain (NEDICES) study in central Spain. We identified all participants with incident PD (N=23), diagnosed in the follow-up examination (1997-1998), who had performed an expanded 37-item version of the Mini-Mental State Examination (37-MMSE) at the baseline evaluation (1994-1995). These 23 were 1:4 matched to 92 controls. RESULTS: Baseline 37-MMSE scores were 27.9+/-4.9 (28) in PD patients and 28.7+/-6.5 (31) in controls (p=0.212). There were no patient-control differences in orientation, immediate recall, attention and calculation, memory recall, language, or visuospatial copying. In analyses that adjusted for several possible confounding factors, there were no case-control differences. CONCLUSIONS: In this population-based sample, patients with incident PD did not have evidence of significant global cognitive function disturbances three years prior to their diagnosis when compared with matched controls. Our data suggest that global cognitive dysfunction does not precede the diagnosis of PD. PMID- 21621226 TI - The importance of non-motor disturbances to quality of life in Parkinson's disease. AB - The term health-related quality of life (HRQoL) refers to the consequences of the disease and treatment on the patients' life, as perceived and evaluated by patients themselves. Parkinson's disease is a complex disorder characterized by motor manifestations and an impressive diversity of non-motor symptoms (NMS) that may be simultaneously present in the same patient. Some of these non-motor symptoms (for example, depression, fatigue, or pain) have got evidence enough to be considered determinant factors of HRQoL. For others, evidence is incomplete or results of the studies are inconsistent. The availability of instruments for assessment of a wide set of NMS allows to evaluate the impact of NMS, as a whole and compared to the motor disorder, on the HRQoL. In that scenario, the whole NMS showed a close association with the quality of life deterioration, exceeding even the effects of the motor disorder. PMID- 21621227 TI - Randomized controlled trial of brief cognitive behavioral intervention for depression and anxiety symptoms preoperatively in patients undergoing coronary artery bypass graft surgery. AB - OBJECTIVE: The goal of this study was to examine the feasibility, acceptability, and efficacy of a brief, tailored cognitive-behavioral intervention for patients with symptoms of preoperative depression or anxiety before undergoing a coronary artery bypass graft (CABG) operation. METHODS: Patients were recruited from a university teaching hospital between February 2007 and May 2009. Patients were randomly assigned to receive treatment as usual (TAU) or a cognitive behavioral therapy (CBT) intervention called Managing Anxiety and Depression using Education and Skills (MADES). A total of 100 subjects were randomized into the study. Length of hospital stay was assessed with a 1-way analysis of variance. Depression, anxiety, and quality of life were assessed with mixed-model repeated measures analyses. RESULTS: Overall, the intervention was feasible, and patients had a positive impression of the MADES. Patients in the TAU group stayed longer in the hospital than did those in the MADES group (7.9 days +/- 2.6 vs 9.2 days +/- 3.5; P = .049). Depressive symptoms increased at time of hospital discharge for the TAU group, whereas the MADES group had a decrease in depressive symptoms at the time of discharge. Quality of life and anxiety symptoms improved in both groups at 3 to 4 weeks of follow-up. However, the MADES group had greater improvements than did the TAU group. CONCLUSIONS: This study demonstrated that brief, tailored CBT targeting preoperative depression and anxiety is both feasible and acceptable for patients undergoing CABG surgery. Most important, this intervention improved depressive and anxiety symptoms, as well as quality of life. Moreover, it reduced in-hospital length of stay. This study found that a cognitive-behavioral intervention for patients undergoing CABG surgery for symptoms of preoperative depression/anxiety is both feasible and acceptable. Most important, this intervention improved depressive and anxiety symptoms, as well as quality of life. It also reduced in-hospital length of stay. PMID- 21621228 TI - [Home intravenous antimicrobial therapy in multi-drug resistant microorganism infections]. AB - BACKGROUND: Although home intravenous antimicrobial infusion therapy (HIVAIT) has proved its safety and efficacy in a great number of common infections, there are few published studies about its role in the treatment of infections caused by multi-drug resistant microorganisms. Our objectives are to study clinical and epidemiological characteristics of patients with multi-drug resistant microorganism infections treated with HIVAIT, and its usefulness in this type of infections. METHODS: We analyzed all patients diagnosed of infections requiring HIVAIT and admitted to our Hospital at Home Unit (HHU) from March 2007 to February 2010. Subjects were divided into two groups: patients with multi-drug resistant microorganism infections as a study group, and the remaining patients as a control group. RESULTS: A total of 487 patients were included, 82 in the study group. Comorbidity and physical dependence were higher in this group than in the control group (p=0.000 and p=0.002 respectively). The majority of patients were discharged because of a satisfactory clinical evolution. However, 17 (20.7%) patients in the study group required readmission to hospital during treatment and another 22 (26.8%) were re-admitted to hospital 3 months after discharge from HHU. There were significant differences between the results from the control group in clinical readmissions. CONCLUSIONS: Patients with multi-drug resistant microorganism infections and HIVAIT have higher comorbidity, physical dependence, and frequency of hospital readmissions. However, HIVAIT is useful in this kind of infections if the patients are appropriately selected. PMID- 21621229 TI - [In-hospital ischemic strokes in patients admitted to Cardiology and Cardiac Surgery departments. Multi-centre registry]. AB - BACKGROUND AND OBJECTIVE: Patients admitted to Cardiology and Cardiac Surgery Departments have an increased risk of ischemic stroke (IS). We analyzed clinical characteristics, quality of neurological care and mortality of in-hospital strokes (IHS) in these departments. PATIENTS AND METHOD: Prospective registry of in-hospital ISs in Cardiology and Cardiac Surgery in 13 Spanish hospitals during 2008. Demographic, clinical and therapeutic data as well as mortality and functional evolution were recorded. RESULTS: 73 patients were included. Mean age was 72+/-11.6 years. 75.4% of IS were cardioembolic. Special risk factors were presence of cardiac sources of embolism (86.3%), prior withdrawal of antithrombotic treatment (22%) and invasive procedures (65.7%). First neurological assessment was done in the first 3hours in 49.5% and beyond 24hours from IS onset in 20.5%. Ten patients were treated with intravenous thrombolysis, which was not possible in 8 patients because of the delay in calling the neurologist. Most frequent reasons for exclusion from thrombolytic therapy were recent major surgical procedures (33.3%) and anticoagulant therapy (38%). Three month mortality was 15% and only 53.7% were functionally independent. Patients treated with thrombolysis had a better evolution (87.5% of independent patients, p=0.04). CONCLUSIONS: IS in Cardiology and Cardiac Surgery are mostly cardioembolic strokes and produce a high proportion of dependent patients. Patients treated with thrombolysis had a better evolution. Delays in contacting the neurologist led to exclusion from treatment an important proportion of patients who met thrombolysis criteria. PMID- 21621230 TI - [Health status in Spain between 2000 and 2009: an approach through the analysis of the press media]. PMID- 21621231 TI - Immune response to Rhodococcus equi ATCC 33701-secreted proteins in mice and identification of immunogenic recombinant proteins by dot-blotting. AB - Rhodococcus equi remains a significant pathogen, causing severe pneumonia in foals. The development of vaccines and serologic diagnosis could be greatly facilitated by studying the humoral immune response to this equine pathogen. In this study, a crude extract of R. equi ATCC 33701-secreted proteins combined with the Montanide(r) ISA70 adjuvant was found to be highly immunogenic in mice with the highest titer of 99,000 on day 42 after the first subcutaneous immunization. This immune response was dependent on the quantity of proteins injected and the presence of adjuvant. By dot-blotting, eight recombinant secreted proteins were identified to react strongly with sera from immunized mice. Of these eight proteins, four were detected as immunogenic only when administered in conjunction with adjuvant. This screening strategy led to the identification of promising new candidates for vaccine development. PMID- 21621232 TI - Assessment of the heat stability of seven rapid HIV assays. AB - Human Immunodeficiency Virus Rapid Diagnostic Tests (HIV RDTs) are robust, quick to perform, effective diagnostic tools. The stability of seven commonly used RDTs for detecting antibody to HIV was assessed during exposure to temperatures above 30 degrees C, the usual maximum recommended by manufacturers. The aim of the study was to determine if HIV RDTs retain their testing outcomes after exposure to higher temperatures. At two testing sites, seven RDTs were exposed to a short heat shock (60 degrees C for 72 hours) as might occur during transport. RDTs were exposed to ambient (22 or 30 degrees C), warm (35 or 37 degrees C) or hot (45 degrees C) temperatures for up to 90 days. Testing was performed at five intervals on a panel of seven positive and three negative plasma samples. Results showed no changes consistent with altered testing outcomes over time and/or temperature when test indicators were compared with the control indicators. Only one HIV RDT achieved 100% consensus with reference results at all four storage temperatures (median 97.5%, lowest 74% for RDT5 at 45 degrees C). Testing outcomes in a limited sample panel showed six of seven HIV RDT kits were relatively robust despite exposure to higher than recommended temperatures. PMID- 21621233 TI - Predictive values of prurigo nodularis and herpes zoster for HIV infection and immunosuppression requiring HAART in French Guiana. AB - Prurigo nodularis and herpes zoster frequently lead to the diagnosis of HIV in tropical areas. The WHO has established a clinical definition of AIDS for undeveloped countries. Prurigo and herpes zoster are both classified as stage 2. The main objective of this study was to compare the level of immunosuppression of patients diagnosed as HIV-positive after consulting for prurigo nodularis or herpes zoster in French Guiana. A retrospective study was conducted including patients consulting at the Department of Dermatology, Cayenne Hospital (French Guiana) for prurigo nodularis or herpes zoster between 1989 and 2007 for which the systematic HIV test was positive. Demographic data and CD4 counts of both groups were compared. Analysis of 346 patients consulting for herpes zoster (n=192) or prurigo nodularis (n=154) led to the discovery of 129 HIV infections. The positive predictive value (PPV) for HIV positivity was 38.5% for herpes zoster and 36% for prurigo nodularis. The median lymphocyte count was 302/mm(3) in herpes zoster and 87/mm(3) in prurigo nodularis (P<0.001). The PPV for having a CD4 lymphocyte count<200/mm(3) was 26.5% for herpes zoster and 72% for prurigo nodularis. Prurigo nodularis was predictive of advanced immunosuppression. This questions the pertinence of the WHO clinical classification of AIDS. In the absence of CD4 count, the present results suggest that for patients with prurigo nodularis, antiretrovirals should be initiated without delay. PMID- 21621234 TI - Activity of Metarhizium spp. and Isaria spp. from the Central Brazilian Cerrado against Triatoma infestans nymphs. AB - Entomopathogenic fungi attack Chagas disease vectors, and there is a need of effective isolates for development of biological control. Ten isolates of Metarhizium spp., four of Isaria cateniannulata and two of an Isaria sp., all isolated with Triatoma infestans as a bait insect from soils collected in Central Brazil, and tested under laboratory conditions were effective against third instar nymphs of T. infestans. This is the first report of pathogenicity of M. flavoviride var. pemphigi, M. robertsii and I. cateniannulata against T. infestans. M. robertsii IP 34 and I. cateniannulata IP 141 are among the most virulent fungi reported, and both have potential for integrated control of triatomine vectors. PMID- 21621235 TI - Liver angiomyolipomas: a clinical, radiologic, and pathologic analysis of 22 patients from a single center. AB - BACKGROUND: Liver angiomyolipomas (AML) are mesenchymal neoplasms with an uncertain clinical behavior. The spectrum of presentation, imaging and histologic features, and outcomes were analyzed in all patients treated at Memorial Sloan Kettering Cancer Center. METHODS: Demographics, disease, pathologic, treatment, and outcome-related variables for consecutive patients were reviewed retrospectively. All imaging studies obtained at presentation were reexamined, categorized, and compared using Fisher and Wilcoxon tests. RESULTS: Between 1989 and 2010, 238 patients with AML were seen and 22 (9.3%) had liver involvement (exclusive = 17; combined with kidney = 5). The median age was 53 years; 18 were females, and 15 had symptoms. AML was not suspected at initial presentation in any patient. Fat-containing neoplasms on imaging were larger (P = .03), with more heterogeneous enhancement compared with fat-poor neoplasms (P = .001), but none of the imaging/histologic features correlated with outcome. Thirteen patients underwent resection (R0 = 9), 4 were observed, 2 received chemotherapy, 2 embolization, and 1 embolization plus intra-arterial chemotherapy. Two patients treated with R0-resection recurred and 2 treated with chemotherapy progressed, but no patient died of AML-related causes. At a median follow-up of 36 months, 7 patients were free of disease, 13 were alive with disease, 1 died of an unrelated cause, and another was lost to follow-up. CONCLUSION: AMLs are rare neoplasms with an indolent course in most patients. Subtypes based on fat content are recognized, but clinical behavior does not seem to be different. Recurrence after resection is not associated with disease-related mortality. Resection may be unnecessary in selected asymptomatic patients if the diagnosis can be established definitively. PMID- 21621236 TI - Preoperative chemoradiation reduces the risk of pancreatic fistula after distal pancreatectomy for pancreatic adenocarcinoma. AB - BACKGROUND: Pancreatic fistula (PF) is a common complication after pancreatectomy. Previous reports indicate that preoperative irradiation decreases the risk of PF after pancreatoduodenectomy. In this context, the impact of preoperative chemoradiation therapy (CRT) on PF formation after distal pancreatectomy is of interest. METHODS: Fifty-eight patients with pancreatic adenocarcinoma who underwent distal pancreatectomy, including 28 patients with preoperative gemcitabine-based CRT and 30 patients without preoperative treatment, were assessed in this study. The incidence and severity of postoperative PF, assessed according to the definition of the International Study Group on Pancreatic Fistula, were compared between the 2 groups. RESULTS: In the CRT group, 86% of patients did not develop PF, whereas grades A and B PF were observed in 1 and 3 patients, respectively. In the non-CRT group, 33% of patients did not develop a PF, whereas grades A and B PF were observed in 9 and 11 patients, respectively. The incidence of clinically significant PF, defined as either grade B or grade C PF, was less in the CRT group (P = .031). The amylase activities in the draining fluid on postoperative days 1 and 3 were both less in the CRT group (P = .003 and P = .006, respectively). CONCLUSION: Preoperative CRT significantly decreases the incidence of PF after distal pancreatectomy, which potentially provides another benefit to patients in addition to its original advantages (ie, locoregional effect and patient selection effect), allowing more opportunities for the immediate initiation of postoperative adjuvant treatment. PMID- 21621237 TI - A prospective study on elective umbilical hernia repair in patients with liver cirrhosis and ascites. AB - BACKGROUND: Patients with both cirrhosis and ascites have a 20% risk of developing umbilical hernia. A retrospective study from our center comparing conservative management of umbilical hernia with elective repair in these patients showed a significant risk of mortality as a result of hernia incarceration in conservatively treated patients. The goal of this study was to assess the safety and efficacy of elective umbilical hernia repair in these patients prospectively. METHODS: Patients with liver cirrhosis and ascites presenting with an umbilical hernia were included in this study. For all patients, the expected time to liver transplantation was more than 3 months, and they did not have a patent umbilical vein in the hernia sac. The following data were collected prospectively for all patients: Child-Pugh-Turcotte (CPT) classification, model for end-stage liver disease (MELD) score, kidney failure, cardiovascular comorbidity, operation-related complications, and duration of hospital stay. Mortality rates were registered in hospital records and verified in government records during follow-up. Mortality rates were registered in hospital records and verified in government records during follow-up. On completion of the study, a retrospective survey was performed to search for any patients who met the study inclusion criteria but were left out of the study cohort. RESULTS: In total, 30 patients (25 males) underwent operation at a mean age of 58 years (standard deviation [SD] +/- 9 years). Of these 30 patients, 6 were classified as CPT grade A (20%), 19 (63%) as grade B, and 5 (17%) as grade C. The patients' median MELD score was 12 (interquartile range [IQR], 8-16). In 10 (33%) of the 30 patients hernia repair was performed with mesh. The median duration of hospital stay was 3 days (IQR, 2-4). None of the patients were admitted to the intensive care unit. Postoperative complications included pneumonia and decompensation of cirrhosis (1 case each,) resulting in prolonged hospital stay for those 2 patients. After a median follow-up period of 25 months (IQR, 14-34), 2 (7%) of the 30 patients died; neither of the deaths were attributable to the umbilical hernia repair. A total of 2 patients suffered recurrence. CONCLUSION: Elective umbilical hernia repair is safe and the preferred approach in cirrhotic patients with ascites. PMID- 21621238 TI - Thyroidectomy and parathyroidectomy in patients with high body mass index are safe overall: analysis of 26,864 patients. AB - BACKGROUND: Obesity is a national epidemic. Prior studies of the impact of body mass index (BMI) on surgical outcomes from cervical endocrine procedures have come from high-volume, single institutions. Our study characterizes the 30-day clinical and economic outcomes in patients with high BMI from a multi institutional database. METHODS: Patients undergoing thyroidectomy or parathyroidectomy in the American College of Surgeons National Surgery Quality Improvement Program, 2005-2008 were categorized into 4 groups BMI based on: normal, overweight, obese, and morbidly obese. Overweight, obese, and morbidly obese patients were compared with patients with normal BMI using a chi(2) test and an analysis of variance. Multivariable linear/logistic regression models were used to adjust for preoperative risk factors. RESULTS: In all, 18,825 patients underwent thyroidectomy. Overweight, obese, and morbidly obese patients were more likely to have total thyroidectomy, substernal thyroids, general anesthesia, operations of greater duration, and an overall or wound complication (all P < .01). On a multivariable analysis, morbidly obese patients had an increased risk for urinary complications (P < .05); obese and morbidly obese patients had an increased risk for overall or wound complications (P < .01); overweight, obese, and morbidly obese patients had operations of greater duration (P < .05). In all, 8,039 patients underwent parathyroidectomy. Overweight, obese, and morbidly obese patients were more likely to have general anesthesia and operations of greater duration (all P < .01). On multivariable analysis, morbidly obese patients had operations of greater duration (P < .05) and more wound complications (P = .05). CONCLUSION: Patients with high BMI seem to require operations of greater duration and sustain more morbidity after cervical endocrine procedures than patients with normal BMI, but these differences may not be clinically significant. Thyroidectomy and parathyroidectomy can be performed safely, with appropriate surgical decision making. PMID- 21621239 TI - Simultaneous grayscale and subharmonic ultrasound imaging on a modified commercial scanner. AB - OBJECTIVE: To demonstrate the feasibility of simultaneous dual fundamental grayscale and subharmonic imaging on a modified commercial scanner. MOTIVATION: The ability to generate signals at half the insonation frequency is exclusive to ultrasound contrast agents (UCA). Thus, subharmonic imaging (SHI; transmitting at f(0) and receiving at f(0)/2) provides improved visualization of UCA within the vasculature via suppression of the surrounding tissue echoes. While this capability has proven useful in a variety of clinical applications, the SHI suppression of surrounding tissue landmarks (which are needed for sonographic navigation) also limits it use as a primary imaging modality. In this paper we present results using a commercial ultrasound scanner modified to allow imaging in both grayscale (f(0)=4.0 MHz) and SHI (f(0)=2.5 MHz, f(0)/2=1.25 MHz) modes in real time. METHODS: A Logiq 9 ultrasound scanner (GE Healthcare, Milwaukee, WI) with a 4C curvilinear probe was modified to provide this capability. Four commercially available UCA (Definity, Lantheus Medical Imaging, North Billerica, MA; Optison, GE Healthcare, Princeton, NJ; SonoVue, Bracco Imaging, Milan, Italy; and Sonazoid, GE Healthcare, Oslo, Norway) were all investigated in vitro over an acoustic output range of 3.34 MPa. In vivo the subharmonic response of Sonazoid was investigated in the portal veins of four canines (open abdominal cavity) and four patients with suspected portal hypertension. RESULTS: In vitro, the four UCA showed an average maximum subharmonic amplitude of 44.1+/-5.4 dB above the noise floor with a maximum subharmonic amplitude of 48.6+/-1.6 dB provided by Sonazoid. The average in vivo maximum signal above the noise floor from Sonazoid was 20.8+/ 2.3 dB in canines and 33.9+/-5.2 dB in humans. Subharmonic amplitude as a function of acoustic output in both groups matched the S-curve behavior of the agent observed in vitro. The dual grayscale imaging provided easier sonographic navigation, while the degree of tissue suppression in SHI mode varied greatly on a case by case basis. CONCLUSIONS: These results demonstrate the feasibility of dual grayscale and SHI on a modified commercial scanner. The ability to simultaneously visualize both imaging modes in real time should improve the applicability of SHI as a future primary clinical imaging modality. PMID- 21621240 TI - Influenza virus is not restricted by tetherin whereas influenza VLP production is restricted by tetherin. AB - Tetherin (ST2/CD317) is a cellular protein that restricts the release from cells of some enveloped viruses including HIV-1. To examine if influenza virus is affected by tetherin, MDCK cells constitutively expressing human tetherin and control MDCK cells were infected with influenza virus. No difference was observed in infectious titers, at 24 h or 48 h post-infection. In contrast, tetherin expression inhibited influenza virus-like particle (VLP) release into the media. Expression of the HIV protein Vpu overcame the tetherin block of influenza virus VLPs. A human tetherin mutant that lacks a C-terminal GPI anchor attachment signal (tetherin-DeltaGPI) was constructed to test if this mutant could be incorporated into the released virus or VLP particles. Whereas tetherin-DeltaGPI was incorporated into influenza VLPs it was not incorporated into influenza virions. Taken together these data suggest that influenza virions may contain a tetherin antagonist. PMID- 21621241 TI - Selenium speciation in acidic environmental samples: application to acid rain soil interaction at Mount Etna volcano. AB - Speciation plays a crucial role in elemental mobility. However, trace level selenium (Se) speciation analyses in aqueous samples from acidic environments are hampered due to adsorption of the analytes (i.e. selenate, selenite) on precipitates. Such solid phases can form during pH adaptation up till now necessary for chromatographic separation. Thermodynamic calculations in this study predicted that a pH<4 is needed to prevent precipitation of Al and Fe phases. Therefore, a speciation method with a low pH eluent that matches the natural sample pH of acid rain-soil interaction samples from Etna volcano was developed. With a mobile phase containing 20mM ammonium citrate at pH 3, selenate and selenite could be separated in different acidic media (spiked water, rain, soil leachates) in <10 min with a LOQ of 0.2 MUg L(-1) using (78)Se for detection. Applying this speciation analysis to study acid rain-soil interaction using synthetic rain based on H(2)SO(4) and soil samples collected at the flanks of Etna volcano demonstrated the dominance of selenate over selenite in leachates from samples collected close to the volcanic craters. This suggests that competitive behavior with sulfate present in acid rain might be a key factor in Se mobilization. The developed speciation method can significantly contribute to understand Se cycling in acidic, Al/Fe rich environments. PMID- 21621242 TI - Significance of analytical parameters for the understanding of natural organic matter in relation to photocatalytic oxidation. AB - In this review, special interest was devoted to provide information on the surrogate parameters expressing both quality and quantity of organic matter for the understanding of the photocatalytic oxidation of humic substances. Detailed investigation was directed to the application of photocatalysis with reference to source, origin and modeling of organic matter. Evaluation of the literature findings emphasizes that organic matter taken from natural waters are site specific and should be characterized in detail to be comparable to other studies. Taking into account the photocatalytic degradation studies of natural organic matter, humic substances, humic acids and fulvic acids in slurry systems, a procedure could be deduced that depends on the selection of a standard model sample with a representative concentration, selection of a standard photocatalyst and dose (e.g., TiO2 Degussa P-25, 0.25 mg mL(-1)), application of standardized reaction conditions such as light intensity, pH, and temperature. Furthermore, standardized filtration step avoiding organic leaching and selection of the most suitable analytical parameter are the crucial points to be considered. The use of such a protocol could form a basis for the determination of "relative degradation efficiency" of any sample containing natural organic matter, humic substances, humic acids and fulvic acids regardless of dependency on source and origin. PMID- 21621243 TI - Toxic effects of chemical pesticides (trichlorfon and dimehypo) on Dunaliella salina. AB - Dunaliella salina, a unicellular green alga of environmental tolerance, was employed as test organism to investigate the toxicity effects of trichlorfon and dimehypo widely used in agriculture and veterinary as pesticides. The influences of trichlorfon and dimehypo on cell growth, beta-carotene level, cell morphology changes, and activities of superoxide dismutase (Sod) and catalase (Cat) were investigated. At the concentrations less than 0.050 g L(-1) trichlorfon or 0.0005 g L(-1) dimehypo, cell responses were similar to control. When treated with 0.075 0.100 g L(-1) trichlorfon or 0.001-0.004 g L(-1) dimehypo, cell growth and beta carotene levels declined at first and then revived. When concentrations were higher than 0.125 g L(-1) trichlorfon or 0.005 g L(-1) dimehypo, both cell growth and beta-carotene levels decreased until they were undetectable. The 10-d IC50 of trichlorfon and dimehypo on D. salina were 0.179 g L(-1) and 0.032 g L(-1). Both pollutants could stimulate the increase of Cat activity at a low concentration. Tolerance of D. salina to trichlorfon was obviously higher than that of dimehypo. PMID- 21621244 TI - Predictors of orbital convergence in primates: a test of the snake detection hypothesis of primate evolution. AB - Traditional explanations for the evolution of high orbital convergence and stereoscopic vision in primates have focused on how stereopsis might have aided early primates in foraging or locomoting in an arboreal environment. It has recently been suggested that predation risk by constricting snakes was the selective force that favored the evolution of orbital convergence in early primates, and that later exposure to venomous snakes favored further degrees of convergence in anthropoid primates. Our study tests this snake detection hypothesis (SDH) by examining whether orbital convergence among extant primates is indeed associated with the shared evolutionary history with snakes or the risk that snakes pose for a given species. We predicted that orbital convergence would be higher in species that: 1) have a longer history of sympatry with venomous snakes, 2) are likely to encounter snakes more frequently, 3) are less able to detect or deter snakes due to group size effects, and 4) are more likely to be preyed upon by snakes. Results based on phylogenetically independent contrasts do not support the SDH. Orbital convergence shows no relationship to the shared history with venomous snakes, likelihood of encountering snakes, or group size. Moreover, those species less likely to be targeted as prey by snakes show significantly higher values of orbital convergence. Although an improved ability to detect camouflaged snakes, along with other cryptic stimuli, is likely a consequence of increased orbital convergence, this was unlikely to have been the primary selective force favoring the evolution of stereoscopic vision in primates. PMID- 21621245 TI - Morphological correlates of tail length in the catarrhine sacrum. AB - Taillessness is a distinctive synapomorphy of the Hominoidea that has implications for interpretation of the locomotor behaviors and phylogenetic affinities of the clade's earliest members. However, difficulties persist in confidently identifying taillessness in the catarrhine fossil record, stemming largely from our limited knowledge of the anatomical features with which the tail is associated. Here, we compare the morphology of the sacrum, the sole bony link between the tail and the rest of the body, among extant tailless hominoids and a broad sample of extant cercopithecoids known to vary in tail length (i.e., 'very short', 'short', and 'long') in order to identify morphological correlates of tail length. We examine three features of the sacrum, including the shape of the sacrum's caudal articular surface (CAS), the sacrocaudal articulation (SCA) angle, and the lateral expansion of the last sacral vertebra's transverse processes. Compared with all other taxa, 'long'-tailed cercopithecoids have significantly more circularly-shaped CASs, more acute SCA angles, and more laterally expanded transverse processes of the last sacral vertebra. Tailless hominoids have significantly more elliptically-shaped CASs and less laterally expanded transverse processes than all tailed cercopithecoids, but in the latter parameter, they only differ significantly from 'long'-tailed cercopithecoids. Cercopithecoids with 'short' and 'very short' tails are intermediate between tailless hominoids and 'long'-tailed cercopithecoids with respect to CAS shape and lateral expansion of the transverse processes. SCA angle distinguishes clearly among all three cercopithecoid tail length groups. The results of this study provide evidence for significant differences in sacral morphology among extant catarrhines known to differ in tail length, and have implications for making inferences about tail length and function in extinct catarrhines. PMID- 21621246 TI - Real-time measurement of UV-inactivated Escherichia coli bacterial particles by electrospray-assisted UVAPS spectrometry. AB - The ultraviolet aerodynamic particle sizer (UVAPS) is a novel commercially available aerosol spectrometer for real-time continuous monitoring of viable bioaerosols, based on fluorescence from living microorganisms. In a previous study, we developed an electrospray-assisted UVAPS using biological electrospray techniques, which have the advantage of generating non-agglomerated single particles by the repulsive electrical forces. With this electrospraying of suspensions containing microorganisms, the analytical system can supply more accurate and quantitative information about living microorganisms than with conventional aerosolization. Using electrospray-assisted UVAPS, we investigated the characteristics of bacterial particles with various viabilities in real-time. Escherichia coli was used as the test microorganism, and its initial viability was controlled by the degree of exposure to UV irradiation. In the stable cone jet domain, the particle size distributions of test bacterial particles remained almost uniform regardless of the degree of UV inactivation. However, the fluorescence spectra of the bacterial particles changed with the degree of UV inactivation. The fluorescence characteristics of UV-inactivated bacterial particles tended to show a similar decline with viability, determined by the sampling and culture method, although the percentage showing fluorescence was higher than that showing viability. PMID- 21621247 TI - An empirical approach to estimate soil erosion risk in Spain. AB - Soil erosion is one of the most important factors in land degradation and influences desertification worldwide. In 2001, the Spanish Ministry of the Environment launched the 'National Inventory of Soil Erosion (INES) 2002-2012' to study the process of soil erosion in Spain. The aim of the current article is to assess the usefulness of this National Inventory as an instrument of control, measurement and monitoring of soil erosion in Spain. The methodology and main features of this National Inventory are described in detail. The results achieved as of the end of May 2010 are presented, together with an explanation of the utility of the Inventory as a tool for planning forest hydrologic restoration, soil protection, erosion control, and protection against desertification. Finally, the authors make a comparative analysis of similar initiatives for assessing soil erosion in other countries at the national and European levels. PMID- 21621248 TI - Influence of compressive strength and applied force in concrete on particles exposure concentrations during cutting processes. AB - The objective of this research was to identify the influence of applied force (AF) and the compressive strength (CS) of concrete on particle exposure concentrations during concrete cutting processes. Five cutting conditions were selected with AF varied between 9.8 and 49 N and CS varied between 2500 and 6000 psi. For each selected cutting condition, the measured total dust concentrations (C(tot)) were used to further determine the corresponding three health-related exposure concentrations of the inhalable (C(inh)), thoracic (C(thor)), and respirable fraction (C(res)). Results show that particle size distribution was consistently in a bimodal form under all selected cutting conditions. An increase in CS resulted in an increase in coarse particle generations leading to an increase in the four measured particle exposure levels. An increase in AF resulted in an increase in exposure concentrations with a higher fraction of fine particles (i.e., C(tho) and C(res)) However, for particle exposure concentrations with a higher fraction of coarse particles (i.e., C(tot) and C(inh)), an increase in AF resulted in an initial increase, followed by a decrease in concentration. Finally, the above inferences were further confirmed through the use of fixed effect models to determine the influence of both CS and AF on the four exposure concentrations. These results provide a reference for industries to initiate appropriate control strategies to reduce the exposure levels encountered by workers. PMID- 21621249 TI - Genetic defects in Portuguese families with inherited protein C deficiency. PMID- 21621250 TI - The influencing factors for clopidogrel-mediated platelet inhibition are assay dependent. AB - BACKGROUND: Influencing factors for clopidogrel-mediated platelet inhibition have only been evaluated by one or two different test systems in the same population so far. Since previous studies revealed poor correlations between the various platelet function tests, the identification of influencing variables for clopidogrel response may vary from one test system to the next. We therefore investigated whether the influencing factors for clopidogrel-mediated platelet inhibition depend on the used assay. PATIENTS AND METHODS: Adenosine diphosphate (ADP)-inducible platelet reactivity was assessed by light transmission aggregometry (LTA), the VerifyNow P2Y12 assay, the vasodilator-stimulated phosphoprotein (VASP) phosphorylation assay, multiple electrode aggregometry (MEA), and the Impact-R in 288 patients after angioplasty and stenting for cardiovascular disease. By univariate and multivariate regression analyses, we evaluated the impact of age >= 75, gender, body mass index (BMI), diabetes, active smoking, hypertension, hyperlipidemia, C-reactive protein, platelet count, creatinine, use of calcium-channel blockers (CCBs), statins, proton pump inhibitors, beta blockers, angiotensin converting enzyme inhibitors, and angiotensin receptor blockers on clopidogrel-mediated platelet inhibition in each test system. RESULTS: None of the independent influencing variables was consistent through all test systems. Only by LTA and the VerifyNow P2Y12 assay, age >= 75 and the use of CCBs were independently associated with higher on treatment platelet reactivity. Only by the VASP assay and MEA, on-treatment platelet reactivity increased linearly with BMI. Further, only by MEA, residual ADP-inducible platelet reactivity increased linearly with platelet count, whereas an increase in platelet count was independently associated with a decrease in ADP inducible platelet activation by the Impact-R. CONCLUSION: The influencing factors for platelet reactivity during clopidogrel therapy are assay-dependent. PMID- 21621251 TI - PROTHROMBINEX((r))-VF (PTX-VF) usage for reversal of coagulopathy: prospective evaluation of thrombogenic risk. AB - INTRODUCTION: Prothrombin complex concentrates (PCCs) are used for the urgent reversal of oral vitamin K antagonists in patients with life-threatening bleeding or prior to urgent procedures/surgery. PCCs offer rapid and complete reversal without the disadvantages of volume overload and adverse reactions seen with fresh frozen plasma (FFP). There is concern about the risk of thrombosis associated with the use PCCs; data on this is limited at present. OBJECTIVES: To determine the incidence of objectively confirmed arterial or venous thromboembolism within 30 days following the administration of PROTHROMBINEX(r) VF (PTX-VF) to acutely reverse a prolonged INR. MATERIALS/METHODS: A prospective observational study was conducted at two teaching hospitals in Auckland, NZ. All patients who received PTX-VF for the acute reversal of prolonged INR were eligible. Baseline patient demographics and reasons for PTX-VF administration were recorded. Patients were reviewed at days 7 and 30, to confirm/exclude thromboembolism or adverse events. RESULTS: 173 patients were enrolled from August 2008 to March 2009. The most frequent indication for reversal was acute bleeding. At 30 days 4.6% (8/173) patients had a definite/probable thrombotic event, and 16.7% had died either due to the presenting bleed (intracranial haemorrhage) or a complication of their presenting complaint (e.g. sepsis, renal failure). CONCLUSIONS: Acute reversal of anticoagulant therapy with PTX-VF is associated with a significant rate of thromboembolism (4.6%) within 30 days. These events can be explained by ongoing cessation of anticoagulant therapy in patients with ongoing risk factors for arterial or venous thrombosis, rather than directly attributable to PTX-VF therapy. PMID- 21621252 TI - Insights into the inhibition of platelet activation by omega-3 polyunsaturated fatty acids: beyond aspirin and clopidogrel. AB - OBJECTIVES: We sought to examine the effects of escalating doses of omega-3 polyunsaturated fatty acid (PUFA) supplements on platelet function using light transmission aggregometry (LTA) and electrophoretic quasi-elastic light scattering technology (EQELS). BACKGROUND: PUFA may inhibit platelet function through fatty acid substitution in the platelet membrane by changing the surface charge density and causing decreased production of thromboxane A2. EQELS can measure platelet surface charge density and determine whether the platelet is in resting or activated state. METHODS: A total of 30 volunteers were divided in 3 groups of 10 as follows: Group A, no antiplatelet agent; Group B, daily aspirin only, and Group C, daily aspirin and clopidogrel. All patients received escalating doses of omega-3PUFA from 1 to 8 g daily over 24 weeks. Platelet function was measured by template bleeding time, LTA, and EQELS at baseline and at 6, 12, 18 and 24 weeks. RESULTS: Mean bleeding time increased in a dose dependent manner with escalating omega-3 PUFA doses. LTA confirmed expected antiplatelet effects of aspirin and clopidogrel, but did not detect any additional antiplatelet effects of omega-3 PUFA. EQELS showed a significant increase in the negative resting platelet charge compared to baseline and an attenuated response to arachidonic acid mediated platelet activation. No bleeding events were observed. CONCLUSIONS: In this pilot study we were able to successfully measure platelet surface charge variation as a measure of omega-3 PUFA effect on platelets. Our results suggest that omega-3 PUFA increase the total platelet surface charge and, therefore, attenuate platelet activation, even among patients taking aspirin or aspirin plus clopidogrel. Further studies are needed to determine the clinical significance of these measured effects and EQELS results. PMID- 21621253 TI - Ultrastructural organization of muscle fiber types and their distribution in the rat superior rectus extraocular muscle. AB - Extraocular muscles (EOMs) are unique as they show greater variation in anatomical and physiological properties than any other skeletal muscles. To investigate the muscle fiber types and to understand better the structure function correlation of the extraocular muscles, the present study examined the ultrastructural characteristics of the superior rectus muscle of rat. The superior rectus muscle is organized into two layers: a central global layer of mainly large-diameter fibers and an outer C-shaped orbital layer of principally small-diameter fibers. Six morphologically distinct fiber types were identified within the superior rectus muscle. Four muscle fiber types, three single innervated fibers (SIFs) and one multiple innervated fiber (MIF), were recognized in the global layer. The single innervated fibers included red, white and intermediate fibers. They differed from one another with respect to diameter, mitochondrial size and distribution, sarcoplasmic reticulum and myofibrillar size. The orbital layer contained two distinct MIFs in addition to the red and intermediate SIFs. The orbital MIFs were categorized into low oxidative and high oxidative types according to their mitochondrial content and distribution. The highly specialized function of the superior rectus extraocular muscle is reflected in the multiplicity of its fiber types, which exhibit unique structural features. The unique ultrastructural features of the extraocular muscles and their possible relation to muscle function are discussed. PMID- 21621254 TI - Epothilone B enhances Class I HLA and HLA-A2 surface molecule expression in ovarian cancer cells. AB - OBJECTIVE: Ovarian cancer is the leading cause of death from gynecologic cancers in the United States. Epothilone B (EpoB), Taxol and vinblastine are anti neoplastic agents that interfere with microtubules and arrest the cell cycle in the G2/M phase. EpoB is being evaluated in phase III clinical trials, and its analogs are currently being used in the treatment of taxane-resistant metastatic breast cancer. Little is known about the effect of these drugs on the immune response to tumors. Cancer cells evade immune recognition by down-regulating HLA Class I expression, allowing escape from immune surveillance and destruction. Our data illustrates the effect of microtubule-interacting agents on HLA Class I and HLA-A2 expression as well as the modulation of cytokine expression in ovarian cancer cells. METHODS: Ovarian cancer cells were treated with different concentrations of drugs. Cell surface expression and mRNA transcription of HLA Class I molecules and HLA-A2 was examined. IFNalpha, IL1beta, IL12 and IL6 mRNA expression was also evaluated upon EpoB treatment. RESULTS: Low-dose EpoB, Taxol and vinblastine greatly increased expression of HLA Class I and HLA-A2 molecules in Hey ovarian cancer cells. EpoB does not modulate HLA expression in drug resistant ovarian cancer cells. The expression of IFNalpha, IL1beta, IL12 and IL6 is also markedly increased upon EpoB treatment. CONCLUSIONS: Nanomolar concentrations of microtubule-interacting agents enhance immune-visibility of ovarian cancer cells by increasing HLA Class I and pro-inflammatory cytokine expression. Immune recognition of tumor cells may be improved. PMID- 21621255 TI - Mast cell activation syndrome: a newly recognized disorder with systemic clinical manifestations. AB - BACKGROUND: Diagnostic criteria for mast cell (MC) activation syndrome have been recently proposed, but clinical studies to validate these criteria are lacking. OBJECTIVE: We sought to determine the clinical manifestations of this newly recognized syndrome in a cohort of patients. METHODS: We prospectively evaluated 18 patients seen at our institution with MC activation syndrome from 2006 to 2009. Patients enrolled had at least 4 of the signs and symptoms of abdominal pain, diarrhea, flushing, dermatographism, memory and concentration difficulties, or headache. Response to treatment with anti-MC mediator medications was assessed based on established criteria. Laboratory tests indicating MC mediator release and histopathology and immunohistochemical studies on gastrointestinal biopsy samples were performed. RESULTS: Ninety-four percent of the patients had abdominal pain, 89% had dermatographism, 89% had flushing, and 72% had the constellation of all 3 symptoms. Patients additionally had headache, diarrhea, and memory and concentration difficulties. All patients had at least 1 positive laboratory test result for an increased MC mediator level. On the basis of the response to treatment criteria, 67% of the patients in the cohort had either a complete or major regression in symptoms while taking medications targeting MC mediators. There was no significant difference in the numbers of intestinal mucosal MCs between our patients and healthy control subjects. CONCLUSION: MC activation syndrome might be the underlying cause of unexplained symptoms when several organ systems are involved, such as the gastrointestinal tract and the skin. It is especially important to be able to recognize the constellation of clinical features because response to anti-MC mediator medications is often excellent. PMID- 21621256 TI - Cosmopolitan and ethnic-specific replication of genetic risk factors for asthma in 2 Latino populations. AB - BACKGROUND: Although Mexicans and Puerto Ricans are jointly classified as "Hispanic/Latino," there are significant differences in asthma prevalence, severity, and mortality between the 2 groups. We sought to examine the possibility that population-specific genetic risks contribute to this disparity. OBJECTIVES: More than 100 candidate genes have been associated with asthma and replicated in an independent population, and 7 genome-wide association studies in asthma have been performed. We compared the pattern of replication of these associations in Puerto Ricans and Mexicans. METHODS: We genotyped Mexican and Puerto Rican trios using an Affymetrix 6.0 GeneChip and used a family-based analysis to test for genetic associations in 124 genes previously associated with asthma. RESULTS: We identified 32 single nucleotide polymorphisms (SNPs) in 17 genes associated with asthma in at least 1 of the 2 populations. Twenty-two of these SNPs in 11 genes were significantly associated with asthma in the combined population and showed no significant heterogeneity of association, whereas 5 SNPs were associated in only 1 population and showed statistically significant heterogeneity. In a gene-based approach 2 additional genes were associated with asthma in the combined population, and 3 additional genes displayed ethnic specific associations with heterogeneity. CONCLUSIONS: Our results show that only a minority of genetic association studies replicate in our population of Mexican and Puerto Rican asthmatic subjects. Among SNPs that were successfully replicated, most showed no significant heterogeneity across populations. However, we identified several population-specific genetic associations. PMID- 21621257 TI - Pluronic-lysozyme conjugates as anti-adhesive and antibacterial bifunctional polymers for surface coating. AB - This paper describes the preparation and characterization of polymer-protein conjugates composed of a synthetic triblock copolymer with a central polypropylene oxide (PPO) block and two terminal polyethylene oxide (PEO) segments, Pluronic F-127, and the antibacterial enzyme lysozyme attached to the telechelic groups of the PEO chains. Covalent conjugation of lysozyme proceeded via reductive amination of aldehyde functionalized PEO blocks (CHO-Pluronic) and the amine groups of the lysine residues in the protein. SDS-PAGE gel electrophoresis together with MALDI-TOF mass spectrometry analysis revealed formation of conjugates of one or two lysozyme molecules per Pluronic polymer chain. The conjugated lysozyme showed antibacterial activity towards Bacillus subtilis. Analysis with a quartz crystal microbalance with dissipation revealed that Pluronic-lysozyme conjugates adsorb in a brush conformation on a hydrophobic gold-coated quartz surface. X-ray photoelectron spectroscopy indicated surface coverage of 32% by lysozyme when adsorbed from a mixture of unconjugated Pluronic and Pluronic-lysozyme conjugate (ratio 99:1) and of 47% after adsorption of 100% Pluronic-lysozyme conjugates. Thus, bifunctional brushes were created, possessing both anti-adhesive activity due to the polymer brush, combined with the antibacterial activity of lysozyme. The coating having a lower degree of lysozyme coverage proved to be more bactericidal. PMID- 21621259 TI - Biochemical responses and DNA damage in red sea bream from coastal Fujian Province, China. AB - Relationships between measures of oxidative stress and adaptive antioxidant defenses were studied in red sea bream (Pagrus major, RSB) caged at eight locations along the Fujian coast, China. Oxidative damage in tissues was investigated by measuring concentrations of protein carbonyl (CP) and reactive oxygen species (ROS), lipid peroxidation (LPO), acetyl cholinesterase activity (AChE), concentration of thiol groups and activities of enzymes associated with antioxidant defense. Magnitudes of antioxidant defense parameters were significantly, positively correlated with oxidative stress. Significantly greater ROS, LPO and CP content was observed in most tissues of RSB caged at three locations relative to those at the other locations. RSB caged at one location, Meizhou, exhibited significantly more DNA damage than did RSB caged at other locations. AChE activities in liver, kidney and muscle were statistically different among locations. Multivariate analyses were used to categorize the environmental quality of different locations. The battery of measures of oxidative stress proved to be a useful integrator of oxidative stresses that could be compared among locations. PMID- 21621258 TI - TRPML: transporters of metals in lysosomes essential for cell survival? AB - Key aspects of lysosomal function are affected by the ionic content of the lysosomal lumen and, therefore, by the ion permeability in the lysosomal membrane. Such functions include regulation of lysosomal acidification, a critical process in delivery and activation of the lysosomal enzymes, release of metals from lysosomes into the cytoplasm and the Ca(2+)-dependent component of membrane fusion events in the endocytic pathway. While the basic mechanisms of lysosomal acidification have been largely defined, the lysosomal metal transport system is not well understood. TRPML1 is a lysosomal ion channel whose malfunction is implicated in the lysosomal storage disease Mucolipidosis Type IV. Recent evidence suggests that TRPML1 is involved in Fe(2+), Ca(2+) and Zn(2+) transport across the lysosomal membrane, ascribing novel physiological roles to this ion channel, and perhaps to its relatives TRPML2 and TRPML3 and illuminating poorly understood aspects of lysosomal function. Further, alterations in metal transport by the TRPMLs due to mutations or environmental factors may contribute to their role in the disease phenotype and cell death. PMID- 21621260 TI - Determination of selected heavy metal concentrations and distribution in a southwestern stream using macrophytes. AB - Since the reduction of the arsenic standard from 50 to 10 MUg L(-1) by the US Environmental Protection Agency in 2006 many small town and rural water municipalities were left with the task of preventing or mitigating arsenic contamination of drinking water supplies. In this study macrophytes and sediments were used to determine the concentration and distribution of heavy metals (As, Cd, Cu, Pb, and Zn) within the primary source of drinking water (Gallinas River watershed) to the residents of Las Vegas, New Mexico. Sampling was done in the spring and fall at four sites, two above the city and two below, and samples were analyzed using ICP-MS. Results showed significantly higher (p<.05) metal concentrations in plant roots than shoots for most metals. Spearman's correlation showed positive correlations (r>.3) between plant and sediment concentrations of Cd, Pb, Zn, As, and a negative correlation for Cu. The site above waste water treatment plant (AWWTP) had the highest plant tissue concentrations of Cd, Pb, Zn, and As. All of these concentrations attained critical toxicity levels exceeding sediment quality guidelines. High concentration factor values and levels of metals detected in macrophyte tissues indicate that heavy metals within sediments in the Gallinas River occur in bioavailable forms. Correlations between plant and sediment metal concentrations indicate that metal concentrations in macrophyte tissues are a good reflection of metal concentrations within the sediment in the Gallinas River. PMID- 21621261 TI - Prevalence and causes of visual impairment and blindness in an urban Indian population: the Singapore Indian Eye Study. AB - PURPOSE: To describe the prevalence and causes of visual impairment and blindness in an urban Indian population. DESIGN: Population-based study. PARTICIPANTS: Ethnic Indians aged more than 40 years living in Singapore. METHODS: Participants underwent standardized ophthalmic assessments for visual impairment and blindness, defined using best-corrected visual acuity (BCVA) and presenting visual acuity (PVA), according to US and modified World Health Organization (WHO) definitions. MAIN OUTCOME MEASURES: Unilateral visual impairment or blindness was defined on the basis of the worse eye, and bilateral visual impairment or blindness was defined on the basis of the better eye. Primary causes of visual impairment were determined. RESULTS: A total of 3400 eligible individuals (75.6% response rate) participated. On the basis of US definitions, the age-standardized prevalence was 0.4% for bilateral blindness (<=20/200, better eye) and 3.4% for bilateral visual impairment (<20/40 to >20/200, better eye). Another 0.3% of bilateral blindness and 13.4% of bilateral visual impairment were correctable with refraction. Cataract was the principal cause of best-corrected bilateral blindness (60.0%) and bilateral visual impairment (65.7%). Other major causes of blindness and visual impairment included diabetic retinopathy, age-related macular degeneration, glaucoma, corneal opacity, and myopic maculopathy. CONCLUSIONS: The prevalence of bilateral blindness and visual impairment in Indians living in Singapore is lower than estimates from populations living in India, but similar to estimates obtained from Singapore Malay and Chinese populations. Cataract is the leading cause of blindness and visual impairment. One in 20 cases of bilateral blindness and 1 in 10 cases of bilateral visual impairment are attributable to diabetic retinopathy. These data may have relevance to many ethnic Indian persons living outside India. PMID- 21621262 TI - Retinal nerve fiber layer thickness in dominant optic atrophy measurements by optical coherence tomography and correlation with age. AB - PURPOSE: To measure the retinal nerve fiber layer (RNFL) thickness by means of optical coherence tomography (OCT) in patients with dominant optic atrophy (DOA). DESIGN: Cross-sectional study. PARTICIPANTS: Thirty-three patients from 15 pedigrees with DOA harboring heterozygous mutations in the OPA1 gene and 43 healthy subjects were enrolled. METHODS: The RNFLs of DOA patients were studied by OCT and compared with those of 43 healthy subjects matched for age and optic nerve head (ONH) size. MAIN OUTCOME MEASURES: Retinal nerve fiber layer thickness. RESULTS: Dominant optic atrophy patients revealed a significant RNFL thickness reduction in all quadrants, with a preferential involvement of the temporal and inferior sectors. The progressive decline of RNFL thickness with age was similar to that observed in healthy subjects and was more evident in the 2 quadrants with higher residual amounts of fibers, that is, the superior and the inferior. The temporal quadrant was profoundly depleted of fiber so that the further rate of loss of microns per year is close to zero, whereas the nasal quadrant was spared the most by neurodegeneration. CONCLUSIONS: The present findings, taken in conjunction with the authors' previous description of small ONH size in DOA, strongly suggest that patients with this disease are born with fewer optic nerve axons and support the hypothesis that subsequent visual loss depends on further age-related loss of fibers, which also occurs in controls. FINANCIAL DISCLOSURE(S): The author(s) have no proprietary or commercial interest in any materials discussed in this article. PMID- 21621263 TI - Striatal and cortical midline activation and connectivity associated with suicidal ideation and depression in bipolar II disorder. AB - BACKGROUND: Considerable evidence implicates dysfunction of striatal and cortical midline structure (CMS) circuitry in mood disorders. Whether such aberrations exist in bipolar II depression is unknown. METHODS: Sixteen unmedicated subjects with bipolar II depression and 19 healthy controls were studied using functional MRI and a motor activation paradigm. Analyses of both activation and functional connectivity were conducted. RESULTS: A history of suicidal ideation (SI) was negatively correlated with activation of the left putamen while depression severity was positively correlated with activation of the left thalamus. The superior bilateral putamen was simultaneously correlated with depression severity and anti-correlated with SI. Striatal functional connectivity was altered with the bilateral CMS and right inferior parietal lobule. Depression severity was correlated with strength of connectivity between the bilateral striatum and the right lingual gyrus and left cerebellum. LIMITATIONS: Only males experiencing an episode of major depression were studied. CONCLUSIONS: Striatal and CMS circuit abnormalities likely contribute to the neurobiology of bipolar II depression. Altered connectivity of the striatum may directly impact depression severity. Further, dissociable components of activation associated with depression severity and suicidal ideation may exist. Finally, the motor activation paradigm used in this study appears to be a useful probe of some neural processes underlying bipolar II depression. PMID- 21621265 TI - Stressful events, stress perception and treatment outcomes in patients with depressive disorders: the CRESCEND study. AB - BACKGROUND: Investigations of associations between stressful events and depression treatment outcomes have led to conflicting findings. In a prospective naturalistic study of depression treatment we sought to investigate perceived stress as a predictor of 12-week antidepressant treatment outcome. METHODS: A nationwide sample of 580 people with depressive disorders was recruited from 18 hospitals in Korea. Number of stressful events in the last 12 months and subjective perception of stress were ascertained, and were dichotomized by median values. Participants commenced on antidepressant treatment were re-evaluated at 1, 2, 4, 8, and 12 weeks later. Assessment scales for evaluating depression (HAMD), anxiety (HAMA), global severity (CGI-s), and functioning (SOFAS) were administered at baseline and at every follow-up visit. Covariates included pre treatment socio-demographic and clinical characteristics, and treatment-related characteristics. RESULTS: Higher baseline perceived stress was significantly associated with worse 12-week antidepressant treatment outcomes in terms of depression, anxiety, and global severity after adjustment for all covariates. However, baseline number of stressful events was not associated with any treatment outcomes. LIMITATIONS: The study was observational, and the treatment modality was naturalistic. CONCLUSIONS: Depressive patients with higher level of perceived stress at the time of commencing treatment had less favorable outcomes after antidepressant treatment. This may represent a group requiring more specific assessment and more intensive management in order to improve treatment response. PMID- 21621266 TI - Bipolarity and inadequate response to antidepressant drugs: clinical and psychopharmacological perspective. AB - OBJECTIVES: The reason why depression may respond poorly to treatment with antidepressant drugs may be connected with the features of bipolarity. Evidence to this effect has accumulated in recent studies of various kinds of depression in mood disorders. Additional evidence for such a connection may be the efficacy of mood-stabilizing drugs in the augmentation of antidepressants in treatment resistant depression. METHODS: This review is based on clinical and psychopharmacological research performed over the past five years. The clinical investigation was based on the response to antidepressants of bipolar depression or to symptoms of hypomania, assessed mainly by the Mood Disorder Questionnaire (MDQ) and the Hypomania Checklist-32 (HCL-32). The psychopharmacological research tested the efficacy of augmentation of antidepressants in treatment-resistant depression by mood-stabilizing drugs of the 1st and 2nd generations. RESULTS: A number of studies have pointed to an association between bipolar depression, or symptoms of hypomania and an inadequate response to antidepressants. Such a connection was also found in the Polish TRES-DEP study which included 1051 depressed patients. Pharmacological studies have demonstrated the efficacy of first generation mood-stabilizing drugs (lithium, carbamazepine) and second generation drugs (quetiapine, olanzapine, risperidone, ziprasidone, lamotrigine) for augmentation of antidepressants in treatment-resistant depression. Some evidence has been presented that mixed depressive episodes may also belong to this category. CONCLUSIONS: The results of these clinical and psychopharmacological studies appear to confirm an association between bipolarity and a poor response of depression to treatment with antidepressant drugs. PMID- 21621264 TI - High and low suicidality in Europe: a fine-grained comparison of France and Spain within the ESEMeD surveys. AB - BACKGROUND: Suicidality risk-factors between countries with similar economic and religious background have been rarely compared, especially within genders. METHODS: Lifetime prevalence of suicide ideation, plans, and attempts in the ESEMeD surveys were stratified on four separate groups: French women, Spanish women, French men, and Spanish men. Outcome odds-ratios (OR) were modelled within each group using logistic regression including demographic characteristics, lifetime mood/anxiety disorders, parental bonding, marital status, and health service-use. RESULTS: Lifetime prevalence of suicide attempts was 3.4% in France (1.1% men, 5.4% women) and 1.5% in Spain (1.2% men, 1.7% women), with a significantly greater gender difference in France (p=0.001). Regarding risk factors, French women reported suicide attempt more commonly with authoritarian mothers (OR=1.51; 95%CI=1.04-2.18), unlike Spanish women (OR=0.77; 95%CI=0.51 1.15) (p<0.001). Spanish men showed more than eight-times higher odds of suicide attempt with overprotecting mothers than French men (p=0.03). General practitioner-(GP)-use was significantly protective of suicide attempt among Spanish women (OR=0.08; 95%CI=0.02-0.35) with no effect in French women (OR=1.03; 95%CI=0.54-2.00) (p=0.01). No significant differences in the effect of marital status, any lifetime antidepressant use, mental disorders, or religiosity on suicide attempt were observed between France and Spain within gender-stratum. LIMITATIONS: Parental bonding is retrospective and potentially influenced by mental state. Response rate was considerably lower in France than in Spain. CONCLUSIONS: Suicidality risk-factors play different roles across genders between France and Spain. Parental bonding dimensions may be interpreted differently according to country, underlining cultural importance. As recommended by WHO, mental health decisions must involve GPs in conjunction with psychiatrists or psychologists. PMID- 21621267 TI - The 2-week duration criterion and severity and course of early childhood depression: implications for nosology. AB - BACKGROUND: Although validity for DSM-IV MDD symptom criteria in preschoolers has been demonstrated, whether the 2-week duration criterion is an appropriate threshold of clinical significance at this age remains unclear. The current study aimed to begin addressing this question. METHOD: Three hundred and six preschoolers were recruited from community sites and followed longitudinally for 2 years. A subsample including healthy preschoolers (N=77) and those with MDD (N=74) were examined. The MDD group was further divided based upon meeting (DSM, N=24) or failing to meet (=65 years; p<0.001) were African American. LVSF was evaluated in 72%, 72%, 70% and 60% (overall p<0.001) and discharge prescriptions of ACE inhibitors or ARBs were given to 83%, 77%, 75% and 75% of eligible patients (overall p=0.013) among those <65, 65-74, 75-84 and >=85 years respectively. During 9 years of follow-up, all-cause mortality occurred in 54%, 61%, 71% and 80% (overall p<0.001) and hospital readmission due to worsening HF occurred in 65%, 60%, 55% and 48% (overall p<0.001) of those <65, 65-74, 75-84 and >=85 years respectively. CONCLUSION: Medicare beneficiaries <65 years with HF, nearly half of whom were African American generally received better quality of care, had lower mortality, but had higher re-hospitalizations due to HF. PMID- 21621286 TI - The large clinical statin trials. PMID- 21621287 TI - Risk factors for delays between intake and veterinary approval for adoption on medical grounds in shelter puppies and kittens. AB - To maximize their capacity to save lives and optimize resource allocation, animal shelters need to identify highly adoptable animals that are unlikely to be delayed on medical grounds before they can be made available for adoption. In this retrospective cohort study, our objective was to identify risk factors for delays from intake to approval for adoption on medical grounds in shelter puppies and kittens. Shelter medical records from 2008 for 335 puppies and 370 kittens were selected randomly at a large metropolitan adoption-guarantee shelter. Data including signalment, source shelter, intake veterinary examination findings, clinical history and days from intake until approval by a veterinarian for adoption on medical grounds were extracted from shelter records and analyzed using multivariate Cox regression. Puppies and kittens with clinical signs of respiratory or gastrointestinal disease at intake took significantly longer to receive approval for adoption on medical grounds (puppies - respiratory p<0.0001; gastrointestinal p<0.0001; kittens - respiratory p<0.0001; gastrointestinal p=0.002). Stray kittens were more likely to be delayed than owner-relinquished kittens or those transferred from other shelters (p<0.01). Older kittens were less likely to be delayed (p<0.0001). Administration of oral or parenteral antibiotics to puppies and kittens with respiratory and/or ocular signs within 24h of intake significantly reduced time to approval on medical grounds for adoption (puppies p=0.02; kittens p=0.03). The analyses suggested that puppies and kittens with respiratory or gastrointestinal signs on intake are more likely to experience delays between intake and veterinary approval for adoption on medical grounds. Prompt antimicrobial treatment of animals with respiratory and/or ocular signs may decrease length of stay in the shelter. PMID- 21621288 TI - Changing mix of medical care services: stylized facts and implications for price indexes. AB - The utilization of health care services has undergone several important shifts in recent years that have implications for the cost of medical care. We empirically document the presence of these shifts for a broad list of medical conditions and assess the implications for price indexes. Following the earlier literature, we compare the growth of two price measures: one that tracks expenditures for the services actually provided to treat conditions and another that holds the mix of those services fixed over time. Using retrospective claims data for a sample of commercially insured patients, we find that, on average, expenditures to treat diseases rose 11% from 2003Q1 to 2005Q4 and would have risen even faster, 18%, had the mix of services remained fixed at the 2003Q1 levels. This suggests that fixed-basket price indexes, as are used in the official statistics, could overstate true price growth significantly. PMID- 21621289 TI - Chemoradiation treatment with gemcitabine plus stereotactic body radiotherapy for unresectable, non-metastatic, locally advanced hilar cholangiocarcinoma. Results of a five year experience. AB - BACKGROUND: Hilar cholangiocarcinoma (Klatskin tumor-KT) accounts for about 0.5 1.5% of all gastrointestinal cancers and for 40-60% of all biliary malignancies. Tumor resection is attainable in about 30-50% of patients. When resection is not possible other treatment options have little or no impact on survival. We present the results of hypofractionated Stereotactic Body Radiotherapy (SBRT) on a small series of non resectable locally advanced KT patients. MATERIALS AND METHODS: Ten patients with histologically proven KT underwent SBRT plus gemcitabine. Radiotherapy (30Gy) was delivered in three fractions. Treatment toxicity was assessed according to the Common Terminology Criteria for Adverse Events (CTCAE v. 3.0). Alive patients with less than 1 year of follow up were excluded from the present study. Local control was assessed according to Response Evaluation Criteria in Solid Tumors (RECIST) criteria. RESULTS: Two grade 1 and Two grade 2 acute toxicities were observed, moreover one grade 2 late toxicity was recorded. The overall local response ratio was 80% (4 PR+2 SD). SBRT showed a good efficacy in achieving local control. Median time to progression was 30 months. Two-year survival was 80% and four-year survival 30%. Six patients developed metastatic disease. Response to treatment and nodal metastases were the only independent indicators of prolonged survival. CONCLUSIONS: The chemoradiation given by SBRT plus gemcitabine is a promising treatment for non-metastatic unresectable KT. High local control rates, even compared to historical data from conventional radiotherapy, can be achieved with minimal toxicity. PMID- 21621290 TI - Optimal diving models: their development and critique requires accurate physiological understanding. PMID- 21621291 TI - Comparison of nonideal sorption formulations in modeling the transport of phthalate esters through packed soil columns. AB - Sorption of dimethyl phthalate (DMP), diethyl phthalate (DEP) and dipropyl phthalate (DPP) to two soil materials that vary in organic matter content was investigated using miscible displacement experiments under saturated flow conditions. Generated breakthrough curves (BTCs) were inversely simulated using linear, equilibrium sorption (LE), nonlinear, equilibrium sorption (NL), linear, first-order nonequilibrium sorption (LFO), linear, radial diffusion (LRD), and nonlinear, first-order nonequilibrium sorption (NFO) models. The Akaike information criterion was utilized to determine the preferred model. The LE model could not adequately describe phthalate ester (PE) BTCs in higher organic matter soil or for more hydrophobic PEs. The LFO and LRD models adequately described the BTCs but a slight improvement in curve-fitting was gained in some cases when the NFO model was used. However, none of the models could properly describe the desorptive tail of DPP for the high organic matter soil. Transport of DPP through this soil was adequately predicted when degradation or sorption hysteresis was considered. Using the optimized parameter values along with values reported by others it was shown that the organic carbon distribution coefficient (K(oc)) of PEs correlates well with the octanol/water partition coefficient (K(ow)). Also, a strong relationship was found between the first-order sorption rate coefficient normalized to injection pulse size and compound residence time. A similar trend of timescale dependence was found for the rate parameter in the radial diffusion model. Results also revealed that the fraction of instantaneous sorption sites is dependent on K(ow) and appears to decrease with the increase in the sorption rate parameter. PMID- 21621292 TI - The phosphoenolpyruvate-dependent glucose-phosphotransferase system from Escherichia coli K-12 as the center of a network regulating carbohydrate flux in the cell. AB - The phosphoenolpyruvate-(PEP)-dependent-carbohydrate:phosphotransferase systems (PTSs) of enteric bacteria constitute a complex transport and sensory system. Such a PTS usually consists of two cytoplasmic energy-coupling proteins, Enzyme I (EI) and HPr, and one of more than 20 different carbohydrate-specific membrane proteins named Enzyme II (EII), which catalyze the uptake and concomitant phosphorylation of numerous carbohydrates. The most prominent representative is the glucose-PTS, which uses a PTS-typical phosphorylation cascade to transport and phosphorylate glucose. All components of the glucose-PTS interact with a large number of non-PTS proteins to regulate the carbohydrate flux in the bacterial cell. Several aspects of the glucose-PTS have been intensively investigated in various research projects of many groups. In this article we will review our recent findings on a Glc-PTS-dependent metalloprotease, on the interaction of EIICB(Glc) with the regulatory peptide SgrT, on the structure of the membrane spanning C-domain of the glucose transporter and on the modeling approaches of ptsG regulation, respectively, and discuss them in context of general PTS research. PMID- 21621293 TI - Membrane proteins--functional dynamics and coupling to cellular networks. PMID- 21621294 TI - Nonablative fractional photothermolysis for facial actinic keratoses: 6-month follow-up with histologic evaluation. AB - BACKGROUND: A number of epidermal and papillary dermal skin conditions can be treated safely and effectively with fractional photothermolysis (FP). OBJECTIVE: We sought to evaluate the effectiveness of FP with a 1550-nm fractionated erbium doped fiber laser for the treatment of facial actinic keratoses (AKs). METHODS: Fourteen men, ages 59 to 79 years, underwent 5 laser treatments (2- to 4-week intervals) at an energy fluence of 20 to 70 mJ and treatment level of 11 (8-10 passes), corresponding to 32% to 40% surface area coverage. AK counts and photographs were taken at baseline, before each treatment, and at 1-, 3-, and 6 month follow-ups after the last treatment. Biopsies were performed at baseline and at the 3-month follow-up. The clinical improvement of the actinic lesions was evaluated by a dermatologist using digital photography and lesion counts at all 3 follow-up visits. RESULTS: The AK count for each patient was reduced on average by 73.1% (67.5%-77.7%) at the 1-month, 66.2% (60.0%-71.5%) at the 3-month, and 55.6% (43.9%-64.8%) at the 6-month follow-up visit. Excluding two cases, all biopsy specimens (baseline and at the 3-month follow-up) were positive for histologic features of AK and/or squamous cell carcinoma. LIMITATIONS: This study is limited by a small number of patients; therefore further clinical studies are warranted. CONCLUSIONS: FP decreases the number of clinical AKs; however, posttreatment biopsy specimens indicate the histologic persistence of AKs (epidermal tumors). FP is not an adequate single-treatment modality for AKs. PMID- 21621295 TI - The efficacy of infection control interventions in reducing the incidence of extended-spectrum beta-lactamase-producing Enterobacteriaceae in the nonoutbreak setting: A systematic review. AB - We conducted a systematic review to examine the efficacy of infection control interventions for the control of ESBL-producing Enterobacteriaceae in hospitals in the non-outbreak setting. Although 4 uncontrolled, retrospective studies were included in the review, no well designed prospective studies capable of informing infection control practice were identified, underscoring the urgent need for research in this area. PMID- 21621296 TI - Changes in acetylcholinesterase expression are associated with altered presenilin 1 levels. AB - We have previously identified presenilin-1 (PS1), the active component of the gamma-secretase complex, as an interacting protein of the amyloid-associated enzyme acetylcholinesterase (AChE). In this study, we have explored the consequences of AChE-PS1 interactions. Treatment of SH-SY5Y cells with the AChE inhibitor tacrine decreased PS1 levels, in parallel with increase in the secretion of amyloid precursor protein APPalpha, whereas the cholinergic agonist carbachol had no effect on PS1. AChE knockdown with siRNA also decreased PS1 levels, while AChE overexpression exerted opposing effect. AChE-deficient also had decreased PS1. Mice administered with tacrine or donepezil displayed lower levels of brain PS1. However, sustained AChE inhibition failed to exert long-term effect on PS1. This limited duration of response may be due to AChE upregulation caused by chronic inhibition. Finally, we exposed SH-SY5Y cells to beta-amyloid (Abeta)42 which triggered elevation of both AChE and PS1 levels. The Abeta42 induced PS1 increase was abolished by siRNA AChE pretreatment, suggesting that AChE may participate in the pathological feedback loop between PS1 and Abeta. Our results provide insight into AChE-amyloid interrelationships. PMID- 21621297 TI - SOD1 G93D sporadic amyotrophic lateral sclerosis (SALS) patient with rapid progression and concomitant novel ANG variant. AB - SOD1 G93D mutation has been described in amyotrophic lateral sclerosis (ALS) patients with slowly progressive disease. We describe an Italian patient affected by sporadic ALS with the SOD1 G93D mutation that disclosed an unusual rapid progression with death occurring after 30 months from the symptom onset. Considering the atypical clinical course further genes associated with ALS or known to be causative were studied including ANG, PGRN, TARDBP, FUS, VCP, CHRNA3, CHRNA4, and CHRNB4. A novel heterozygous ANG missense variant (c.433 C>T, p.R145C) was identified which is neither reported in controls nor in 1000 genomes and single nucleotide polymorphism (SNP) databases. This report confirms that clinical course of SOD1-related ALS may be modulated by other causative or associated genes, including ANG and suggests that extensive screening of ALS associated genes in patients with an already identified mutation may be helpful for better knowledge of genetic architecture of ALS. PMID- 21621298 TI - A meta-analysis of diffusion tensor imaging studies in amyotrophic lateral sclerosis. AB - Studies involving diffusion tensor imaging (DTI) of amyotrophic lateral sclerosis (ALS) with whole-brain voxel-based analysis yielded variable findings. A systematic review was conducted on whole-brain voxel-based diffusion tensor imaging fractional anisotropy (FA) studies of ALS patients and healthy controls (HC) in PubMed, ISI Web of Science, Embase, and MEDLINE databases from 1990 to December 25, 2010. Coordinates were extracted from clusters with significant difference in FA between ALS patients and HC. Meta-analysis was performed using signed differential mapping. Eight studies were enrolled, comprising 143 ALS patients and 145 HC. The included studies reported FA reduction at 67 coordinates in ALS and no FA increased. Significant reductions were present in the bilateral frontal white matter/cingulate gyrus and the posterior limb of bilateral internal capsule. The findings remain largely unchanged in quartile and jackknife sensitivity analyses. Our finding suggests that ALS is a multisystem disease beyond motor dysfunction and provides evidence that FA reduction in the frontal white matter and cingulate gyrus may be a special biomarker of ALS. PMID- 21621299 TI - No association of toll-like receptor 2 polymorphisms with Alzheimer's disease in Han Chinese. AB - Toll-like receptor 2 (TLR2) represents a reasonable functional and positional candidate gene for Alzheimer's disease (AD) as it is located under the linkage region of AD on chromosome 4q, and is functionally involved in the microglia mediated inflammatory response and amyloid beta (Abeta) clearance. In the current study, 7 single nucleotide polymorphisms (SNPs) that span the TLR2 were selected and their associations with late-onset AD (LOAD) risk were assessed in a case control sample comprising 785 individuals in a Han Chinese population. No significant differences in the frequency of TLR2 alleles, genotypes, and haplotypes in the AD cases were detected compared with the controls. TLR2 gene might not play a major role in the genetic predisposition to late-onset Alzheimer's disease in this population. PMID- 21621300 TI - Modeling hippocampal neurogenesis across the lifespan in seven species. AB - The aim of this study was to estimate the number of new cells and neurons added to the dentate gyrus across the lifespan, and to compare the rate of age associated decline in neurogenesis across species. Data from mice (Mus musculus), rats (Rattus norvegicus), lesser hedgehog tenrecs (Echinops telfairi), macaques (Macaca mulatta), marmosets (Callithrix jacchus), tree shrews (Tupaia belangeri), and humans (Homo sapiens) were extracted from 21 data sets published in 14 different reports. Analysis of variance (ANOVA), exponential, Weibull, and power models were fit to the data to determine which best described the relationship between age and neurogenesis. Exponential models provided a suitable fit and were used to estimate the relevant parameters. The rate of decrease of neurogenesis correlated with species longevity (r = 0.769, p = 0.043), but not body mass or basal metabolic rate. Of all the cells added postnatally to the mouse dentate gyrus, only 8.5% (95% confidence interval [CI], 1.0% to 14.7%) of these will be added after middle age. In addition, only 5.7% (95% CI 0.7% to 9.9%) of the existing cell population turns over from middle age and onward. Thus, relatively few new cells are added for much of an animal's life, and only a proportion of these will mature into functional neurons. PMID- 21621301 TI - [Hexaminolevulinate photodynamic diagnosis in non-muscle invasive bladder cancer: experience of the BLUE group]. AB - OBJECTIVES: Photodynamic diagnosis (PDD) with hexaminolevulinate has been recently used to improve detection of non-muscle invasive bladder cancer. Our main purpose was to quantify the benefit of PDD vs. conventional white light cystoscopy (WL) in our area. MATERIAL AND METHODS: Fluorescence-guided cystoscopy using hexaminolevulinate was performed at the time of the transurethral resection (TUR) in 305 patients from 7 Spanish hospitals. All lesions found with WL and PDD were numbered and recorded in an online database. Each lesion was sent separately for pathology analysis. Random biopsies were also obtained in 148 patients. RESULTS: A total of 1659 lesions were biopsied: 522 were identified with PDD and WL, 237 only with PDD, 19 only with WL and 881 random biopsies. Of the 600 tumors, PDD detected 563, WL 441 and random biopsies 29 (20 CIS). The mean overdetection rate for PDD over WL was 31.9% for all types of lesions, but it was 209% for carcinoma in situ (CIS). Sensitivity was 93.8% for PDD and 78.2% for WL. Specificity was 81.5% for PDD and 90.5% for WL. In 23% of patients, PDD detected at least one additional neoplastic lesion compared to WL. CONCLUSIONS: Hexaminolevulinate fluorescence cystoscopy improves detection and resection of non-muscle invasive bladder cancer, especially of CIS. Sensitivity of PDD is higher than WL, but specificity is lower. In our study, random biopsies were able to detect some CIS not visible under PDD. PMID- 21621302 TI - [Estimation of the projections of incidence rates, prevalence and mortality due to prostate cancer in Aragon (Spain)]. AB - OBJECTIVE: The purpose of this study was to project prostate cancer prevalence, incidence rates and mortality over a 15-year period going from 2008 to 2022 for the Aragon region of Spain. METHODS: The projections were calculated using the MIAMOD PROGRAM. All-cause mortality due to prostate cancer for the baseline period of 1998 to 2007 was obtained from the Spanish National Institute of Statistics. RESULTS: The prostate cancer incidence rates for the period 2008 to 2022 per 100,000 persons are projected to decrease from 47.57 to 26.68/100,000 (adjusted rates 24.9 to 12.93). Mortality will decrease from 21.89 to 11.73 (adjusted rates 10.89 to 5.83) and prevalence from 277.07 cases/ 100,000 per year to 125.09 (adjusted rates 165.54 to 63.41). CONCLUSIONS: These projections indicate that the prostate cancer incidence and its mortality rate will decrease in Aragon (Spain). PMID- 21621303 TI - [Association of nocturnal penile rigidity with testosterone, metabolic syndrome, and other variables: a prospective cross-sectional pilot study]. AB - INTRODUCTION: The aim was to study whether nocturnal penile rigidity (NPTR) correlates with metabolic syndrome (MetS) and testosterone in men consulting for erectile dysfunction (ED). MATERIAL AND METHODS: 234 men were included in a prospective, cross-sectional pilot study. Serum total and bioavailable testosterone and other biochemical constituents were measured and compared with NPTR. Patients were classified by normal or low/abnormal penile rigidity (abnormal meaning predominant organic component of ED) and presence or absence of MetS to test the hypothesized correlations. RESULTS: Application of the logistic regression model to rigidity as the dependent variable showed the risk of low penile rigidity to be significantly lower for patients with higher total (OR=0.96, 95% CI=0.92-0.99) or bioavailable testosterone (OR=0.91, 95% CI=0.84 0.99). Patients with testosterone levels between 8 and 12 mmol/L had a quadrupled risk of low penile rigidity compared with patients with higher levels (>12 mmol/L) (OR=3.96, 95% CI=1.89-8.31). Considering men without MetS, age and body mass index were associated as significant factors for low penile rigidity: age increased risk by 8% (OR=1.08, 95% CI=1.03-1.13) and BMI increased it by 18% (OR=1.18, 95% CI=1.01-1.38). CONCLUSION: Testosterone levels are weakly associated with penile rigidity and disappear when associated with MetS. PMID- 21621304 TI - [Efficacy and safety of remifentanil analgesia in the immediate postoperative period of maxillofacial surgery]. PMID- 21621305 TI - [Threatening hemoptysis and pulmonary vein stenosis after ablation due to atrial fibrillation]. PMID- 21621306 TI - [Sternal osteomyelitis and cellulitis after percutaneous endoscopic gastrostomy]. PMID- 21621307 TI - [Management of COPD patients in a health centre]. PMID- 21621308 TI - [Miller-Fisher syndrome associated with acute motor axonal neuropathy: clinic immunological correlation]. PMID- 21621309 TI - [Meningeal siderosis. A case study and a literature review]. PMID- 21621310 TI - Evaluation of doctor/patient satisfaction with the use of the Parkinson's Disease Dementia-Short-Screen (PDD-SS): a screening test for dementia in Parkinson's disease (DIFFUSION study). AB - INTRODUCTION: Parkinson disease (PD) has no specific neuropsychological scales for assessing the most significant cognitive impairment in PD, which has determined the use of subjective criteria or instruments designed for other diseases, making difficult the comparison between studies or the follow-up of patients. A screening test for dementia in PD (Parkinson's Disease Dementia-Short Screen [PDD-SS]) has recently been validated. To assess the degree of satisfaction of patients and researchers through the use of PDD-SS in clinical practice. PATIENTS AND METHODS: An observational, cross-over, multicentre and national study was conducted on 471 patients with PD. The degree of patient satisfaction was measured using a questionnaire in which the items scored from 0 to 10 on a visual analogue scale (0 = strongly disagree, 10 = completely agree), while the researchers were determined on a 1-5 point Likert scale (1 = strongly disagree, 5 = completely agree). RESULTS: A total of 171 patients (36.3%) patients had dementia associated with PD according to the PDD-SS, of whom 77.3% said they were satisfied with its use. The overall measurement of researcher satisfaction was 3.6+/-0.6 points. Ninety per cent (n=45) of them reported an overall score >3 points in the satisfaction questionnaire. The mean values of perception of applicability, usability and reliability of PDD-SS among researchers was 3.5+/-0.7, 3.7+/-0.6 and 3.1+/-0.5 points, respectively. CONCLUSIONS: PD patients, as well as most of the researchers, were satisfied with the use of PDD-SS in clinical practice. PMID- 21621311 TI - Comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) of some benzimidazole derivatives with trichomonicidal activity. AB - Trichomonosis is a common sexually transmitted infectious disease linked to reproductive health complications. Recently, the benzimidazole nucleus has emerged as a promising scaffold to develop new trichomonicidal agents. Despite the fact that large amounts of experimental data have been accumulated over the past eight years, no quantitative studies have yet been reported on this class of compounds. In our effort to develop new antiparasitic benzimidazole derivatives, we report in this paper CoMFA and CoMSIA studies with an initial set of 70 benzimidazole derivatives with trichomonicidal activity. Four CoMFA models and eight CoMSIA models were generated; ten of these models had values of r(2) > 0.6 and q(2) > 0.5. The best CoMFA model had r(2) = 0.936 and q(2) = 0.634, and the best CoMSIA model had r(2) = 0.858 and q(2) = 0.642. These models were generated by using two conformer selection methodologies (minimum energy conformations and 3D similarity), and three charge types (Mulliken, Gasteiger-Hukel and electrostatic potential atomic charges). The putative active tautomers of 1H benzimidazole derivatives were selected using 3D-QSAR calculations. All models were validated via an external test set with 13 molecules. The best models satisfied additional validation criteria. The contour maps generated show the most important features that a benzimidazole derivative should have for trichomonicidal activity; they also, suggest that substituents at the 2- and 6 positions are important in the generation of derivatives with strong activity. PMID- 21621312 TI - 8-Aryl- and alkyloxycaffeine analogues as inhibitors of monoamine oxidase. AB - Recently it was reported that a series of 8-benzyloxycaffeine analogues are potent reversible inhibitors of human monoamine oxidase (MAO) A and B. In an attempt to discover additional C8 oxy substituents of caffeine that lead to potent MAO inhibition, a series of related 8-aryl- and alkyloxycaffeine analogues were synthesized and their MAO-A and -B inhibition potencies were compared to those of the 8-benzyloxycaffeines. The results document that while the 8 substituted-oxycaffeine analogues inhibited both human MAO isoforms, they displayed a high degree of selectivity for MAO-B. 8-(3-Phenylpropoxy)caffeine, 8 (2-phenoxyethoxy)caffeine and 8-[(5-methylhexyl)oxy]caffeine were found to be the especially potent MAO-B inhibitors with IC(50) values ranging from 0.38 to 0.62 MUM. These inhibitors are therefore 2.5-4.6 fold more potent MAO-B inhibitors than is 8-benzyloxycaffeine (IC(50) = 1.77 MUM). It is also demonstrated that, analogous to 8-benzyloxycaffeine, halogen substitution on the phenyl ring of the C8 substituent significantly enhances MAO binding affinity. For example, the most potent MAO-B inhibitor of the present series is 8-[2-(4 bromophenoxy)ethoxy]caffeine with an IC(50) value of 0.166 MUM. This study also reports possible binding orientations of selected oxy caffeines within the active site cavities of MAO-A and MAO-B. PMID- 21621313 TI - The stability of an alar cinch suture after Le Fort I and mandibular osteotomies in Japanese patients with Class III malocclusions. AB - We previously reported a modified technique for the placement of symmetrical cinch sutures after switching from a nasal to an oral endotracheal tube. We undertook a study to assess the effectiveness of our technique and the stability of changes in the nasolabial morphology after bimaxillary surgery. The study group comprised 30 patients aged 17-36 years who had skeletal Class III malocclusion. All patients had bimaxillary surgery with an alar base cinch suture and V-Y closure. The nasal region was measured directly or on cephalograms before, and 1 week and 1 year after operation. The suture did not alter the width of the alar base, but the nasolabial angle and projection of the tip increased significantly. The length of the upper lip did not change significantly. PMID- 21621314 TI - Accumulation of mercury and methylmercury by mushrooms and earthworms from forest soils. AB - Accumulation of total and methyl-Hg by mushrooms and earthworms was studied in thirty-four natural forest soils strongly varying in soil physico-chemical characteristics. Tissue Hg concentrations of both receptors did hardly correlate with Hg concentrations in soil. Both total and methyl-Hg concentrations in tissues were species-specific and dependent on the ecological groups of receptor. Methyl-Hg was low accounting for less than 5 and 8% of total Hg in tissues of mushrooms and earthworms, respectively, but with four times higher concentrations in earthworms than mushrooms. Total Hg concentrations in mushrooms averaged 0.96 mg Hg kg(-1) dw whereas litter decomposing mushrooms showed highest total Hg and methyl-Hg concentrations. Earthworms contained similar Hg concentrations (1.04 mg Hg kg(-1) dw) whereas endogeic earthworms accumulated highest amounts of Hg and methyl-Hg. PMID- 21621315 TI - Outcomes following military traumatic cardiorespiratory arrest: A prospective observational study. AB - AIM: To determine the characteristics of military traumatic cardiorespiratory arrest (TCRA), and to identify factors associated with successful resuscitation. METHODS: Data was collected prospectively for adult casualties suffering TCRA presenting to a military field hospital in Helmand Province, Afghanistan between 29 November 2009 and 13 June 2010. RESULTS: Data was available for 52 patients meeting the inclusion criteria. The mean age (range) was 25 (18-36) years. The principal mechanism of injury was improvised explosive device (IED) explosion, the lower limbs were the most common sites of injury and exsanguination was the most common cause of arrest. Fourteen (27%) patients exhibited ROSC and four (8%) survived to discharge. All survivors achieved a good neurological recovery by Glasgow Outcome Scale. Three survivors had arrested due to exsanguination and one had arrested due to pericardial tamponade. All survivors had arrested after commencing transport to hospital and the longest duration of arrest associated with survival was 24 min. All survivors demonstrated PEA rhythms on ECG during arrest. When performed, 6/24 patients had ultrasound evidence of cardiac activity during arrest; all six with cardiac activity subsequently exhibited ROSC and two survived to hospital discharge. CONCLUSION: Overall rates of survival from military TCRA were similar to published civilian data, despite military TCRA victims presenting with high Injury Severity Scores and exsanguination due to blast and fragmentation injuries. Factors associated with successful resuscitation included arrest beginning after transport to hospital, the presence of electrical activity on ECG, and the presence of cardiac movement on ultrasound examination. PMID- 21621316 TI - Change of mean platelet volume values in asthmatic children as an inflammatory marker. AB - BACKGROUND: Asthma is the most common chronic disease of childhood in industrialised countries. T helper-2 (Th-2) cells, mast cells and eosinophils have a role in inflammation of asthma. Recently it was shown that platelets also play a role in asthma. Mean platelet volume shows platelet size and reflects platelet activation. OBJECTIVE: The aim of this retrospective study is to evaluate levels of mean platelet volume in asthmatic patients during asymptomatic periods and exacerbations compared with healthy controls. METHODS: The study consisted of 100 asthmatic patients (male/female: 55/45, mean age: 8.2+/-3.3) and 49 age and sex matched healthy children as a control group. RESULTS: Mean platelet volume values of asthmatic patients during asymptomatic period were 7.7+/-0.8fL while mean platelet volume values in asthmatics during exacerbation were 7.8+/-0.9fL. Comparison of mean platelet volume values of asthmatic patients and healthy controls both in acute asthmatic attack and asymptomatic period showed no difference (p>0.05). Comparison of mean platelet volume values at asthmatic attack and asymptomatic period also had no difference (p>0.05). The presence of atopy, infection, eosinophilia, elevated immunoglobulin E, and severity of acute asthmatic attack did not influence mean platelet volume values. CONCLUSION: The results of our study suggest that mean platelet volume values may not be used as a marker in bronchial asthma, although prospective studies with larger number of patients are needed to evaluate the role of mean platelet volume in asthma. PMID- 21621317 TI - Cefotaxim induced a near fatal anaphylactic shock in an infant. PMID- 21621318 TI - Risks associated with laparoscopic entry: guidelines for clinical practice from the French College of Gynaecologists and Obstetricians. AB - The aim of these recommendations of the French National College of Gynaecologists and Obstetricians was to focus the surgeon's attention on those aspects which could allow him/her to prevent, or at least limit, the incidence of these serious complications, in the absence of a previous laparotomy or specific risk factors (obesity, gauntness, large pelvic mass or pregnancy), four widely evaluated techniques can be used in a first line approach (Grade B): blind trans-umbilical technique following creation of pneumoperitoneum with a needle, open laparoscopy (Hasson technique), left upper quadrant entry (pneumoperitoneum and insertion of the first trocar) and direct trans-umbilical trocar with no prior pneumoperitoneum. The currently existing trials do not allow one or another of these techniques to be preferred. Radially expanding insertion systems and optical trocars cannot be recommended as a first-line approach, as a consequence of their currently insufficient degree of evaluation (Grade C). Trans-umbilical (blind or open) laparoscopic entry in a slim woman must be associated with care, as a result of the proximity of the large vessels (Grade B). If a blind trans umbilical insertion technique is decided upon, one option can be to insufflate into the left upper quadrant (professional consensus). In the case of a previous midline laparotomy, whatever the technique used, initial entry is recommended at a distance from the scars (Grade B). It is recommended to carry out micro laparoscopy in the LUQ, because this is the most completely evaluated technique for this indication (Grade C). One option is to use open laparoscopy at a distance from the existing scars (professional consensus). During pregnancy, the insertion position of the first laparoscopic trocar will need to be adapted according to the volume of the uterus (Grade B). Starting from 14WG, trans umbilical Veress needle insufflation is contraindicated (Grade C). Two trocar insertion techniques are thus recommended: open laparoscopy (using the trans umbilical or supra-umbilical routes, depending on the volume of the uterus) or micro-laparoscopy via the left upper quadrant (Grade C). After the second quarter of pregnancy, with laparoscopy the patient will need to be placed on a table inclined towards her left side, in order to minimize compression of the inferior vena cava (Grade B). In the case of laparoscopy during pregnancy, the insufflation pressure must be maintained at a maximum of 12mmHg (Grade B). After 24WG, if laparoscopy is performed, it is recommended to apply open laparoscopy, above the level of the umbilicus (professional consensus). Patients must be informed of the risks inherent to the insertion of trocars during laparoscopy (vascular, bowel or bladder injury) (Grade B). The more benign the pathology requiring an operation, the more detailed the supplied information must be, including that concerning rare but serious complications (Grade B). PMID- 21621319 TI - Conformational dynamics promote binding diversity of dynein light chain LC8. AB - A highly conserved and ubiquitous protein known as LC8 binds over twenty different partners, characteristic of a molecular hub (Barbar, 2008 Biochemistry, 47, 503-508). Structural studies of LC8 complexes with binding partners having diverse recognition sequences show that the same binding groove of LC8 accommodates the various binding motifs. Here we use thermodynamics and dynamics measurements of LC8 complexes to group LC8 binding partners in two categories: those whose binding is enthalpically driven and those that are entropically favored. Peptides that are enthalpically driven completely silence the millisecond-microsecond relaxation signal, suggesting a significant rigidifying of the binding groove, while peptides in the entropically favored group exhibit the same conformational dynamics as the free protein, suggesting that the peptide sits loosely in the binding groove and so retains flexibility of the groove, and presumably of the bound peptide. The inherent disorder in the LC8 binding groove and in LC8 binding partners allows both types of binding, accounts for the lack of a conserved recognition consensus motif and underlies the binding specificity and broad selectivity observed in LC8 binding. PMID- 21621320 TI - Re: David R. Yates, Morgan Roupret, Marc-Olivier Bitker, Christophe Vaessen. To infinity and beyond: the robotic toy story. Eur Urol 2011;60:263-5. PMID- 21621321 TI - Single-incision mini-slings versus standard midurethral slings in surgical management of female stress urinary incontinence: a meta-analysis of effectiveness and complications. AB - CONTEXT: Single-incision mini-slings (SIMS) have been introduced for the treatment of female stress urinary incontinence (SUI); however, concerns have been raised regarding their efficacy. No systematic reviews or meta-analyses have previously assessed these relatively new procedures. OBJECTIVE: To assess the current evidence of effectiveness and safety of SIMS compared with standard midurethral slings (SMUS) (retropubic and transobturator tension-free vaginal tapes) in the management of female SUI. EVIDENCE ACQUISITION: We conducted a literature search from 1996 to January 2011. Meta-analysis of all randomised controlled trials (RCTs) comparing SIMS versus SMUS was performed in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement. Data were analysed using Rev-Man 5. Primary outcomes were patient reported and objective cure rates. Secondary outcomes included perioperative complications, quality of life (QoL) changes, and costs to health services. EVIDENCE SYNTHESIS: A total of 758 women in nine RCTs with a mean follow-up of 9.5 mo were included. The mean age (52.3 vs 52.1 yr), body mass index (27.4 vs 27.7), and parity (2.4 and 2.4) were comparable for both groups. SIMS were associated with significantly lower patient-reported and objective cure rates at 6-12 mo compared with SMUS (risk ratio [RR]: 0.83; 95% confidence interval [CI], 0.70-0.99, and RR: 0.85; 95% CI, 0.74-0.97, respectively). SIMS were associated with significantly shorter operative time (weighed mean difference [WMD]: 8.67 min; 95% CI, 17.32 to -0.02), lower day 1 pain scores (WMD: 1.74; 95% CI, -2.58 to -0.09), and less postoperative groin pain (RR: 0.18; 95% CI, 0.04-0.72). Repeat continence surgery (RR: 6.72; 95% CI, 2.39-18.89) and de novo urgency incontinence (RR: 2.08; 95% CI, 1.01-4.28) were significantly higher in the SIMS group. There was no significant difference in the QoL scores between the groups (WMD: 33.46; 95% CI, -20.62 to 87.55). No studies compared cost to health services. CONCLUSIONS: SIMS are associated with inferior patient-reported and objective cure rates on the short-term follow-up, as well as higher reoperation rates for SUI when compared with SMUS. PMID- 21621322 TI - Re: Hendrik van Poppel, Luigi da Pozzo, Walter Albrecht, et al. A prospective, randomised EORTC intergroup phase 3 study comparing the oncologic outcome of elective nephron-sparing surgery and radical nephrectomy for low-stage renal cell carcinoma. Eur Urol 2011;59:543-52. PMID- 21621323 TI - Re: Pascal Zehnder, Beat Roth, Frederic Birkhauser, et al. A prospective randomised trial comparing the modified HM3 with the MODULITH((r)) SLX-F2 lithotripter. Eur Urol 2011;59:637-44. PMID- 21621324 TI - Mini-slings for female stress urinary incontinence: not yet at the age of reason. PMID- 21621325 TI - A null-mutation in the Znt7 gene accelerates prostate tumor formation in a transgenic adenocarcinoma mouse prostate model. AB - Decrease of cellular zinc in the epithelium of the prostate has been implicated in the development of prostate cancer. To investigate whether ZnT7, a zinc transporter involved in intracellular zinc accumulation, played a role in prostate cancer development, we generated and characterized a transgenic adenocarcinoma mouse prostate (TRAMP) model with a Znt7-null genetic background. TRAMP mice (TRAMP/Znt7(-/-) and TRAMP/Znt7(+/+)) were euthanized at 6, 8, 16, and 28 weeks of age for histopathological analysis of the prostates and for the presence of prostate tumors and metastasis. At 6 and 8 weeks of age, TRAMP/Znt7( /-) mice displayed higher frequencies of low grade prostatic intraepithelial neoplasia (PIN) and high grade PIN, respectively, in the prostates than the age matched TRAMP/Znt7(+/+) mice. At 16 weeks of age, 33% TRAMP/Znt7(-/-) mice had prostate tumors and one half of the mice with prostate tumors had tumor metastasized to the draining lymph nodes while no prostate tumor was detected in the control TRAMP mice. By 28 weeks, 67% TRAMP/Znt7(-/-) mice developed prostate tumors while only 22% control TRAMP mice had prostate tumors. Furthermore, apoptosis was reduced in the prostates of TRAMP/Znt7(-/-) mice. In conclusion, a null-mutation of the Znt7 gene accelerates prostate tumor formation in TRAMP mice. PMID- 21621326 TI - Thymosin beta 4 induces colon cancer cell migration and clinical metastasis via enhancing ILK/IQGAP1/Rac1 signal transduction pathway. AB - Thymosin beta(4) (Tbeta(4)) overexpression increases cell migration and tumor metastasis. Hence, understanding the mechanism of cancer cell migration induced by Tbeta(4) may provide means to inhibit their metastasis. We demonstrated higher Rac1 activities and expression levels of IQGAP1 and ILK in highly migrated Tbeta(4)-overexpressing SW480 cells. In addition, IQGAP1 formed a complex with ILK and knockdown of Tbeta(4) simultaneously reduced ILK and IQGAP1 protein levels as well as their migration ability. These findings suggest that Tbeta(4) increases migration of colon cancer cells via activating Rac1 by elevating IQGAP1/ILK complexes and IHC results illustrated a similar mechanism occurring in vivo. PMID- 21621327 TI - Characteristics and the behavior in electrostatic precipitators of high-alumina coal fly ash from the Jungar power plant, Inner Mongolia, China. AB - In China, flue gases emitted by coal-fired power plants are mainly cleaned using electrostatic precipitators (ESPs). However, based on observations, there is a decrease in the collection efficiency of ESPs in some power plants after burning Jungar coal in Inner Mongolia. In order to find the mechanism of coal fly ash escaping from ESPs, the size distribution, resistivity, and cohesive force of particulate matter samples from Jungar coal-fired power plants in China were measured using a Bahco centrifuge, a dust electrical resistivity test instrument, and a cohesive force test apparatus invented by the authors. Experiments were carried out to determine the chemical composition and current-voltage curve of fly ash under operating ESPs. The Al(2)O(3) content in fly ash was found to reach more than 50%, with the size distribution showing a higher content of PM2.5 and PM10 in high-alumina coal fly ash than in other coal fly ashes. The resistivity of high-alumina coal fly ash was recorded at over 10(12)Omega cm, but this did not result in a clear back corona. The cohesive force of high-alumina coal fly ash was very little. It was sensitive to smoke speed in the electric field, facilitating dust re-entrainment. PMID- 21621328 TI - Phenanthrene sorption to Chinese coal: importance of coal's geochemical properties. AB - Phenanthrene (Phen) was chosen as the probe compound for determining the sorption of PAHs to a series of different coal samples from China. Based on elemental analysis and nuclear magnetic resonance (NMR) spectra analysis, coal samples were characterized with different metamorphic evolutional degrees. The experimental sorption data were fitted well by the Freundlich model, suggesting enhanced sorption capacity and strong nonlinearity of coal samples. The combined partition and adsorption model yielded a better fit than the Freundlich isotherm, indicating that adsorption dominated the sorption at low aqueous concentrations. Correlations between coal properties and sorption capacity values indicated that C%, H/C and O/C atomic ratios were the key factors controlling the sorption behavior. Compared to total carbon, BC might play more important role in the sorption of Phen to coal samples. Moreover, there existed nonlinear relationships between combined carbon, aromatic and aliphatic carbon contents and log K(Fr) and n values, respectively, indicating the significance of aromatic and aliphatic carbon in the coal sorption behavior. PMID- 21621329 TI - Identification of the heat shock protein 70 (HLHsp70) in Haemaphysalis longicornis. AB - A Haemaphysalis longicornis heat shock protein 70 (HLHsp70) was identified from a cDNA library synthesized from tick eggs. The HLHsp70 cDNA is 2311 bp in length and encodes 661 amino acid residues with the predicted molecular weight of 72.5 kDa and an isoelectronic point (pI) of 5.2. It also contains the highly conserved functional motifs of the Hsp70 family and a specific endoplasmic reticulum (ER) retention signal "KDEL" that is common among ER-localized proteins. The HLHsp70 exhibits 90% amino acid identity to the putative Hsp70 of Ixodes scapularis, and 85% to Gallus gallus 78 kDa glucose-regulated protein precursor. Real time RT-PCR analysis showed that the expression levels of the Hsp70 in ovaries and salivary glands were significantly higher than in other tested tissues in partially fed females. Although the expression level of the HLHsp70 was constantly low in unfed ticks, it was significantly induced by blood-feeding. Further, the expression was positively correlated to the temperature (4-37 degrees C, tested). Western blot analysis showed that the rabbit antiserum against the recombinant HLHsp70 protein (rHLHSP70) recognized bands of approximately 100, 72, and 28 kDa from egg lysates, as well as a 72kDa fragment in protein extracts from partially fed larvae. Immunization of rabbits with the rHLHSP70 did not result in a statistically significant reduction of female tick engorgement and oviposition. These results suggest that although HLHSP70 plays a role in the physiological activities of ticks, as a constitutive protein it was not suitable for selection as a candidate vaccine antigen against ticks. PMID- 21621330 TI - Electrical flash burns due to switchboard explosions in New South Wales--a 9-year experience. AB - PURPOSE: To document the incidence and outcome of flash burns due to electrical switchboard explosions presenting to Concord Hospital Burns Unit, from January 2000 to December 2008. METHODS: The Concord Hospital Burns Unit Database was reviewed for admissions due to electrical burns from January 2000 to December 2008. RESULTS: There were 119 electrical burns admitted during the study period, 20 of which were due to high voltage current. Ninety-nine others were low voltage injuries and included 37 cases of electrical burns due to low voltage electrical switchboard explosions. All of the electrical switchboard burns occurred in male electricians. Twenty-one of the 37 low voltage injuries required admission and 7 of them required skin grafting. The mean LOS was 9.95 days. Twenty cases suffered serious complications including major psychological problems and ocular injuries. CONCLUSIONS: Flash burns resulting from switchboard explosions account for a significant proportion of all electrical burns presenting to our institution. These burns may highlight deficits in taking safety precautions and the use of personal protection equipment. Despite the small area of injury the long term psychological sequelae were significant resulting in a delayed return to employment, and there was a high incidence of eye injuries. Additional efforts are therefore required towards the prevention of such injuries. PMID- 21621331 TI - Prolactin in response to acute psychosocial stress in healthy men and women. AB - BACKGROUND: Serum levels of the anterior pituitary hormone prolactin have been reported to increase in response to different types of psychological stressors in humans. However, experimental laboratory stress studies investigating the acute response of prolactin to psychological stress show inconsistent results as increased, as well as decreased or unchanged levels of prolactin have been reported. OBJECTIVE: The aim of this study was to investigate the effect of acute psychosocial stress on serum concentrations of prolactin in healthy men and women and possible sex differences. METHOD: Thirty men and 15 women (age 30-50 years) underwent Trier Social Stress Test (TSST), a tool for investigating psychobiological stress responses in a laboratory setting. Blood samples were collected before and directly after the stress test and after 30 min of recovery. RESULTS: We observed significantly elevated prolactin levels - along with significantly increased plasma adrenocorticotropic hormone (ACTH), serum cortisol, heart rate, systolic blood pressure (SBP), and diastolic blood pressure (DBP) - in response to the stressor. The prolactin response pattern did not differ between men and women, but there was some indication that women might have higher magnitude of response. Large individual differences regarding the magnitude of response were seen in general. The magnitude of the prolactin response was significantly related to the magnitude of the response of the hypothalamic-pituitary-adrenal (HPA) axis and, to some extent, the cardiovascular responses, indicating that individual differences in prolactin response in healthy men and women are dependent on the general physiological stress activation. In women, the magnitude of response was also related to estradiol level. CONCLUSION: Prolactin does increase in response to psychosocial stress, however, with large individual variation in magnitude of response. The pattern of prolactin response does not differ between men and women. However, there was some indication that women might have higher magnitude of increase than men, and that the magnitude of response in women was dependent on estradiol levels, and this needs to be further studied. PMID- 21621332 TI - Circadian cortisol, depressive symptoms and neurological impairment in early multiple sclerosis. AB - OBJECTIVE: There is evidence for the existence of a hyperactive hypothalamus pituitary-adrenal (HPA) axis and its potential role in disease progression in multiple sclerosis (MS). Depressive symptoms are also common in MS. At the same time, depressive symptoms are often associated with an elevated circadian cortisol secretion. So far, little is known about the interplay between depressive symptoms and circadian HPA axis abnormalities in MS. METHODS: Here we investigated depressive symptoms, circadian HPA axis function, cortisol awakening response (CAR) and neurological impairment in 32 early stage relapsing-remitting MS (RRMS) patients and 16 age- and sex-matched controls. Saliva cortisol samples were collected in patients' home environment. Depressive symptoms were assessed by self-report measures. Neurological impairment was assessed by the Kurtzke Expanded Disability Status Scale (EDSS). RESULTS: RRMS patients expressed a significantly higher CAR when compared to healthy controls. After patients were divided into two groups based on their depressive symptom load (Beck Depression Inventory (BDI); median-split), only RRMS patients with moderately elevated depression scores (BDI high) statistically differed in their cortisol release when compared to healthy controls. RRMS patients with low depression scores (BDI low) expressed similar circadian patterns as healthy controls. Neurological impairment (EDSS) was more pronounced in the BDI high group than in the BDI low group. CONCLUSION: In summary, there is evidence, that a hyperactive HPA axis is primarily present in MS patients expressing moderately elevated depressive symptoms. MS patients with only few depressive symptoms do not significantly differ in CAR when compared to healthy controls. To the best of our knowledge, this is the first study showing that in early stage MS, a hyperactive HPA axis is primarily present in patients who express moderate depressive symptoms. PMID- 21621333 TI - Psychological distress, cortisol stress response and subclinical coronary calcification. AB - OBJECTIVES: Poor mental health has been associated with coronary heart disease (CHD). One hypothesized underlying mechanism is hypothalamus pituitary adrenal axis dysfunction. We examined the associations between psychological distress, cortisol response to laboratory-induced mental stress and subclinical coronary artery calcification (CAC). PARTICIPANTS: 527 volunteers free of CHD (mean age=63.0 +/- 5.7 years), drawn from the Whitehall II cohort. MEASURES: CAC was measured using electron beam computed tomography. Current distress at time of the heart scan was indicated by a Short Form-36 mental health score, whereas long term distress was based on the averaged scores of six assessments over the 15 preceding years. Salivary cortisol was measured in response to mental stressors (Stroop, mirror tracing). RESULTS: Detectable CAC was found in 56.4% (mild/moderate: 46.9%; severe: 9.5%) of the sample. After adjustment for sociodemographics and conventional risk factors, long-term but not current psychological distress was associated with a higher risk of severe CAC (OR per SD increase=1.49, 95%CI=1.03-2.16). Psychological distress was not significantly associated with cortisol stress response. A trend for interaction (p=.09) indicated that individuals with long-term poor mental health and high cortisol reactivity showed the highest odds for severe CAC. CONCLUSIONS: Long-term but not current psychological distress is associated with severe CAC in healthy older subjects. Although psychological distress generally was not associated with cortisol stress responses, participants with both long-term distress and increased cortisol response were especially at risk for severe calcification. PMID- 21621334 TI - Dimensionality of hallucinogen and inhalant/solvent abuse and dependence criteria: implications for the Diagnostic and Statistical Manual of Mental Disorders-Fifth Edition. AB - BACKGROUND: Prior research has demonstrated the dimensionality of Diagnostic and Statistical Manual of Mental Disorders-Fourth Edition (DSM-IV) alcohol, nicotine, cannabis, cocaine and amphetamine abuse and dependence criteria. The purpose of this study was to examine the dimensionality of hallucinogen and inhalant/solvent abuse and dependence criteria. In addition, we assessed the impact of elimination of the legal problems abuse criterion on the information value of the aggregate abuse and dependence criteria, another proposed change for DSM-IV currently lacking empirical justification. METHODS: Factor analyses and item response theory (IRT) analyses were used to explore the unidimisionality and psychometric properties of hallucinogen and inhalant/solvent abuse and dependence criteria using a large representative sample of the United States (U.S.) general population. RESULTS: Hallucinogen and inhalant/solvent abuse and dependence criteria formed unidimensional latent traits. For both substances, IRT models without the legal problems abuse criterion demonstrated better fit than the corresponding model with the legal problem abuse criterion. Further, there were no differences in the information value of the IRT models with and without the legal problems abuse criterion, supporting the elimination of that criterion. No bias in the new diagnoses was observed by sex, age and race-ethnicity. CONCLUSION: Consistent with findings for alcohol, nicotine, cannabis, cocaine and amphetamine abuse and dependence criteria, hallucinogen and inhalant/solvent criteria reflect underlying dimensions of severity. The legal problems criterion associated with each of these substance use disorders can be eliminated with no loss in informational value and an advantage of parsimony. Taken together, these findings support the changes to substance use disorder diagnoses recommended by the DSM-V Substance and Related Disorders Workgroup, that is, combining DSM-IV abuse and dependence criteria and eliminating the legal problems abuse criterion. PMID- 21621335 TI - Omega-3 fatty acids affected human perception of ground beef negatively. AB - The objective was to determine the impact of increasing levels of Eicosapentaenoic acid (EPA; C20:5n3) and Docosahexaenoic acid (DHA; C22:6n3) on beef palatability. Two commercial supplements of EPA and DHA were added to 85% lean ground beef patties (176.2 +/- 3.76 g) with different levels (0, 0.3, 0.4, 0.5, 0.6, 0.7, and 1% as-is). Olive oil was added so that all patties contained 1% added lipid. A control treatment was also prepared with no supplement or olive oil. Sensory evaluation and fatty acid analysis was conducted. Ground beef flavor decreased linearly (P<0.001) with increasing levels of EPA and DHA. Off-aroma and off-flavor increased linearly and then plateaued with increasing levels of EPA (P<0.0001). In conclusion, EPA had a greater negative impact on beef palatability than DHA. Also, the panelists were more sensitive to EPA in off-flavor perception than off-aroma. PMID- 21621336 TI - Protein carbonyls in meat systems: a review. AB - Protein oxidation (P-OX) is an innovative topic of increasing interest among meat researchers. Carbonylation is generally recognized as one of the most remarkable chemical modifications in oxidized proteins. In fact, the quantification of protein carbonyls by the dinitrophenylhydrazine (DNPH) method is the most common procedure for assessing P-OX in meat systems. Numerous studies have investigated the occurrence of protein carbonylation right after slaughter and during subsequent processing and cold storage of meat. However, the significance of protein carbonylation in meat systems is still poorly understood. Beyond their role as markers of protein oxidation, specific protein carbonyls such as alpha aminoadipic and gamma-glutamic semialdehydes (AAS and GGS, respectively) are active compounds that may be implicated in several chemical reactions with relevant consequences on meat quality. The formation of protein carbonyls from particular amino acid side chains contribute to impair the conformation of myofibrillar proteins leading to denaturation and loss of functionality. Recent studies also highlight the potential impact of specific protein carbonyls in particular meat quality traits such as water-holding capacity (WHC), texture, flavor and its nutritional value. As a truly emerging topic, the results from current studies provide grounds from the development of further investigations. The present paper reviews the current knowledge on the mechanisms and consequences of protein carbonylation in meat systems and aims to encourage meat researchers to accomplish further investigations on this fascinating research topic. PMID- 21621337 TI - Phase I results of vinorelbine with concurrent radiotherapy in elderly patients with unresectable, locally advanced non-small-cell lung cancer: West Japan Thoracic Oncology Group (WJTOG3005-DI). AB - PURPOSE: To investigate the safety and efficacy of concurrent vinorelbine and thoracic radiotherapy in elderly patients with locally advanced non-small-cell lung cancer (NSCLC). METHODS AND MATERIALS: Eligible patients were 71 years of age or older with unresectable Stage III NSCLC. Patients were treated with thoracic radiotherapy (60 Gy) and concurrent vinorelbine (20 mg/m(2) in Level 1 and 25 mg/m(2) in Level 2) on Days 1 and 8 every 3 weeks for two cycles, followed by adjuvant vinorelbine (25 mg/m(2)) on Days 1 and 8 every 3 weeks for two cycles. RESULTS: Four patients were enrolled at Level 1. One patient experienced Grade 3 febrile neutropenia at Level 1 and the dose was escalated to Level 2. At Level 2, 2 of 6 patients experienced dose-limiting toxicities (Grade 4 neutropenia in 1 patient and Grade 3 infection in another). Three of 6 patients developed late Grade 2 or 3 pneumonitis. Therefore, the dose was de-escalated to Level 1. An additional 6 patients were enrolled at Level 1, 4 of whom experienced dose-limiting toxicities (incomplete radiotherapy because of Grade 2 pneumonitis in 1 patient and Grade 3 infection in 1, Grade 3 febrile neutropenia in 1, and Grade 3 esophagitis in 1). Moreover, late Grade 3 pneumothorax and Grade 5 pneumonitis occurred in 1 and 1 patient, respectively. Overall, Grade 2, 3 and 5 pneumonitis occurred in 3, 3, and 1 among 16 patients, respectively. CONCLUSIONS: Concurrent vinorelbine and thoracic radiotherapy resulted in a high incidence of severe pneumonitis when the standard dose of this agent was used for elderly patients. We therefore recommend caution in the use of this regimen and schedule for elderly patients. PMID- 21621338 TI - Novel assessment of renal motion in children as measured via four-dimensional computed tomography. AB - OBJECTIVES: Abdominal intensity-modulated radiation therapy and proton therapy require quantification of target and organ motion to optimize localization and treatment. Although addressed in adults, there is no available literature on this issue in pediatric patients. We assessed physiologic renal motion in pediatric patients. METHODS AND MATERIALS: Twenty free-breathing pediatric patients at a median age of 8 years (range, 2-18 years) with intra-abdominal tumors underwent computed tomography simulation and four-dimensional computed tomography acquisition (slice thickness, 3 mm). Kidneys and diaphragms were contoured during eight phases of respiration to estimate center-of-mass motion. We quantified center of kidney mass mobility vectors in three dimensions: anteroposterior (AP), mediolateral (ML), and superoinferior (SI). RESULTS: Kidney motion decreases linearly with decreasing age and height. The 95% confidence interval for the averaged minima and maxima of renal motion in children younger than 9 years was 5 9 mm in the ML direction, 4-11 mm in the AP direction, and 12-25 mm in the SI dimension for both kidneys. In children older than 9 years, the same confidence interval reveals a widening range of motion that was 5-16 mm in the ML direction, 6-17 mm in the AP direction, and 21-52 mm in the SI direction. Although not statistically significant, renal motion correlated with diaphragm motion in older patients. The correlation between diaphragm motion and body mass index was borderline (r = 0.52, p = 0.0816) in younger patients. CONCLUSIONS: Renal motion is age and height dependent. Measuring diaphragmatic motion alone does not reliably quantify pediatric renal motion. Renal motion in young children ranges from 5 to 25 mm in orientation-specific directions. The vectors of motion range from 5 to 52 mm in older children. These preliminary data represent novel analyses of pediatric intra-abdominal organ motion. PMID- 21621339 TI - Dosimetric and clinical analysis of spatial distribution of the radiation dose in gamma knife radiosurgery for vestibular schwannoma. AB - OBJECTIVES: We investigated variations in the distribution of radiation dose inside (dose inhomogeneity) and outside (dose falloff) the target volume during Gamma Knife (GK) irradiation of vestibular schwannoma (VS). We analyzed the relationship between some parameters of dose distribution and the clinical and radiological outcome of patients. METHODS AND MATERIALS: Data from dose plans of 203 patients treated for a vestibular schwannoma by GK C using same prescription dose (12 Gy at the 50% isodose) were collected. Four different dosimetric indexes were defined and calculated retrospectively in all plannings on the basis of dose volume histograms: Paddick conformity index (PI), gradient index (GI), homogeneity index (HI), and unit isocenter (UI). The different measures related to distribution of the radiation dose were compared with hearing and tumor outcome of 203 patients with clinical and radiological follow-up of minimum 2 years. RESULTS: Mean, median, SD, and ranges of the four indexes of dose distribution analyzed were calculated; large variations were found between dose plans. We found a high correlation between the target volume and PI, GI, and UI. No significant association was found between the indexes of dose distribution calculated in this study and tumor control, tumor volume shrinkage, hearing worsening, loss of functional hearing, or complete hearing loss at last follow up. CONCLUSIONS: Parameters of distribution of the radiation dose during GK radiosurgery for VS can be highly variable between dose plans. The tumor and hearing outcome of patients treated is not significantly related to these global indexes of dose distribution inside and around target volume. In GK radiosurgery for VS, the outcome seems more to be influenced by local radiation dose delivered to specific structures or volumes than by global dose gradients. PMID- 21621340 TI - Dentalmaps: automatic dental delineation for radiotherapy planning in head-and neck cancer. AB - PURPOSE: To propose an automatic atlas-based segmentation framework of the dental structures, called Dentalmaps, and to assess its accuracy and relevance to guide dental care in the context of intensity-modulated radiotherapy. METHODS AND MATERIALS: A multi-atlas-based segmentation, less sensitive to artifacts than previously published head-and-neck segmentation methods, was used. The manual segmentations of a 21-patient database were first deformed onto the query using nonlinear registrations with the training images and then fused to estimate the consensus segmentation of the query. RESULTS: The framework was evaluated with a leave-one-out protocol. The maximum doses estimated using manual contours were considered as ground truth and compared with the maximum doses estimated using automatic contours. The dose estimation error was within 2-Gy accuracy in 75% of cases (with a median of 0.9 Gy), whereas it was within 2-Gy accuracy in 30% of cases only with the visual estimation method without any contour, which is the routine practice procedure. CONCLUSIONS: Dose estimates using this framework were more accurate than visual estimates without dental contour. Dentalmaps represents a useful documentation and communication tool between radiation oncologists and dentists in routine practice. Prospective multicenter assessment is underway on patients extrinsic to the database. PMID- 21621341 TI - Predictive models for pulmonary function changes after radiotherapy for breast cancer and lymphoma. AB - PURPOSE: To propose multivariate predictive models for changes in pulmonary function tests (DeltaPFTs) with respect to preradiotherapy (pre-RT) values in patients undergoing RT for breast cancer and lymphoma. METHODS AND MATERIALS: A prospective study was designed to measure DeltaPFTs of patients undergoing RT. Sixty-six patients were included. Spirometry, lung capacity (measured by helium dilution), and diffusing capacity of carbon monoxide tests were used to measure lung function. Two lung definitions were considered: paired lung vs. irradiated lung (IL). Correlation analysis of dosimetric parameters (mean lung dose and the percentage of lung volume receiving more than a threshold dose) and DeltaPFTs was carried out to find the best dosimetric predictor. Chemotherapy, age, smoking, and the selected dose-volume parameter were considered as single and interaction terms in a multivariate analysis. Stability of results was checked by bootstrapping. RESULTS: Both lung definitions proved to be similar. Modeling was carried out for IL. Acute and late damage showed the highest correlations with volumes irradiated above ~20 Gy (maximum R(2) = 0.28) and ~40 Gy (maximum R(2) = 0.21), respectively. RT alone induced a minor and transitory restrictive defect (p = 0.013). Doxorubicin-cyclophosphamide-paclitaxel (Taxol), when administered pre-RT, induced a late, large restrictive effect, independent of RT (p = 0.031). Bootstrap values confirmed the results. CONCLUSIONS: None of the dose-volume parameters was a perfect predictor of outcome. Thus, different predictor models for DeltaPFTs were derived for the IL, which incorporated other nondosimetric parameters mainly through interaction terms. Late DeltaPFTs seem to behave more serially than early ones. Large restrictive defects were demonstrated in patients pretreated with doxorubicin-cyclophosphamide-paclitaxel. PMID- 21621342 TI - Anterior myocardial territory may replace the heart as organ at risk in intensity modulated radiotherapy for left-sided breast cancer. AB - PURPOSE: We investigated whether the heart could be replaced by the anterior myocardial territory (AMT) as the organ at risk (OAR) in intensity-modulated radiotherapy (IMRT) of the breast for patients with left-sided breast cancer. METHODS AND MATERIALS: Twenty-three patients with left-sided breast cancer who received postoperative radiation after breast-conserving surgery were studied. For each patient, we generated five IMRT plans including heart (H), left ventricle (LV), AMT, LV+AMT, and H+LV as the primary OARs, respectively, except both lungs and right breast, which corresponded to IMRT(H), IMRT(LV), IMRT(AMT), IMRT(LV+AMT), and IMRT(H+LV). For the planning target volumes and OARs, the parameters of dose-volume histograms were compared. RESULTS: The homogeneity index, conformity index, and coverage index were not compromised significantly in IMRT(AMT), IMRT(LV) and IMRT(LV+ AMT), respectively, when compared with IMRT(H). The mean dose to the heart, LV, and AMT decreased 5.3-21.5% (p < 0.05), 19.9 29.5% (p < 0.05), and 13.3-24.5% (p < 0.05), respectively. Similarly, the low (e.g., V5%), middle (e.g., V20%), and high (e.g., V30%) dose-volume of the heart, LV, and AMT decreased with different levels. The mean dose and V10% of the right lung increased by 9.2% (p < 0.05) and 27.6% (p < 0.05), respectively, in IMRT(LV), and the mean dose and V5% of the right breast decreased significantly in IMRT(AMT) and IMRT(LV+AMT). IMRT(AMT) was the preferred plan and was then compared with IMRT(H+LV); the majority of dose-volume histogram parameters of OARs including the heart, LV, AMT, both lungs, and the right breast were not statistically different. However, the low dose-volume of LV increased and the middle dose-volume decreased significantly (p < 0.05) in IMRT(AMT). Also, those of the right lung (V10%, V15%) and right breast (V5%, V10%) decreased significantly (p < 0.05). CONCLUSIONS: The AMT may replace the heart as the OAR in left-sided breast IMRT after breast-conserving surgery to decrease the radiation dose to the heart. PMID- 21621343 TI - Long-term quality of life outcome after proton beam monotherapy for localized prostate cancer. AB - OBJECTIVES: High-dose external radiation for localized prostate cancer results in favorable clinical outcomes and low toxicity rates. Here, we report long-term quality of life (QOL) outcome for men treated with conformal protons. METHODS: QOL questionnaires were sent at specified intervals to 95 men who received proton radiation. Of these, 87 men reported 3- and/or 12-month outcomes, whereas 73 also reported long-term outcomes (minimum 2 years). Symptom scores were calculated at baseline, 3 months, 12 months, and long-term follow-up. Generalized estimating equation models were constructed to assess longitudinal outcomes while accounting for correlation among repeated measures in an individual patient. Men were stratified into functional groups from their baseline questionnaires (normal, intermediate, or poor function) for each symptom domain. Long-term QOL changes were assessed overall and within functional groups using the Wilcoxon signed-rank test. RESULTS: Statistically significant changes in all four symptom scores were observed in the longitudinal analysis. For the 73 men reporting long-term outcomes, there were significant change scores for incontinence (ID), bowel (BD) and sexual dysfunction (SD), but not obstructive/irritative voiding dysfunction (OID). When stratified by baseline functional category, only men with normal function had increased scores for ID and BD. For SD, there were significant changes in men with both normal and intermediate function, but not poor function. CONCLUSIONS: Patient reported outcomes are sensitive indicators of treatment related morbidity. These results quantitate the long-term consequences of proton monotherapy for prostate cancer. Analysis by baseline functional category provides an individualized prediction of long-term QOL scores. High dose proton radiation was associated with small increases in bowel dysfunction and incontinence, with more pronounced changes in sexual dysfunction. PMID- 21621344 TI - Vasculatures in tumors growing from preirradiated tissues: formed by vasculogenesis and resistant to radiation and antiangiogenic therapy. AB - PURPOSE: To investigate vasculatures and microenvironment in tumors growing from preirradiated tissues (pre-IR tumors) and study the vascular responses of pre-IR tumors to radiation and antiangiogenic therapy. METHODS AND MATERIALS: Transgenic adenocarcinoma of the mouse prostate C1 tumors were implanted into unirradiated or preirradiated tissues and examined for vascularity, hypoxia, and tumor associated macrophage (TAM) infiltrates by immunohistochemistry. The origin of tumor endothelial cells was studied by green fluorescent protein-tagged bone marrow (GFP-BM) transplantation. The response of tumor endothelial cells to radiation and antiangiogenic agent was evaluated by apoptotic assay. RESULTS: The pre-IR tumors had obvious tumor bed effects (TBE), with slower growth rate, lower microvascular density (MVD), and more necrotic and hypoxic fraction compared with control tumors. The vessels were dilated, tightly adhered with pericytes, and incorporated with transplanted GFP-BM cells. In addition, hypoxic regions became aggregated with TAM. As pre-IR tumors developed, the TBE was overcome at the tumor edge where the MVD increased, TAM did not aggregate, and the GFP-BM cells did not incorporate into the vessels. The vessels at tumor edge were more sensitive to the following ionizing radiation and antiangiogenic agent than those in the central low MVD regions. CONCLUSIONS: This study demonstrates that vasculatures in regions with TBE are mainly formed by vasculogenesis and resistant to radiation and antiangiogenic therapy. Tumor bed effects could be overcome at the edge of larger tumors, but where vasculatures are formed by angiogenesis and sensitive to both treatments. Vasculatures formed by vasculogenesis should be the crucial target for the treatment of recurrent tumors after radiotherapy. PMID- 21621345 TI - Comparison of the effects of carbon ion and photon irradiation on the angiogenic response in human lung adenocarcinoma cells. AB - PURPOSE: Radiotherapy resistance is a commonly encountered problem in cancer treatment. In this regard, stabilization of endothelial cells and release of angiogenic factors by cancer cells contribute to this problem. In this study, we used human lung adenocarcinoma (A549) cells to compare the effects of carbon ion and X-ray irradiation on the cells' angiogenic response. METHODS AND MATERIALS: A549 cells were irradiated with biologically equivalent doses for cell survival of either carbon ions (linear energy transfer, 170 keV/MUm; energy of 9.8 MeV/u on target) or X-rays and injected with basement membrane matrix into BALB/c nu/nu mice to generate a plug, allowing quantification of angiogenesis by blood vessel enumeration. The expression of angiogenic factors (VEGF, PlGF, SDF-1, and SCF) was assessed at the mRNA and secreted protein levels by using real-time reverse transcription-PCR and enzyme-linked immunosorbent assay. Signal transduction mediated by stem cell factor (SCF) was assessed by phosphorylation of its receptor c-Kit. For inhibition of SCF/c-Kit signaling, a specific SCF/c-Kit inhibitor (ISCK03) was used. RESULTS: Irradiation of A549 cells with X-rays (6 Gy) but not carbon ions (2 Gy) resulted in a significant increase in blood vessel density (control, 20.71 +/- 1.55; X-ray, 36.44 +/- 3.44; carbon ion, 16.33 +/- 1.03; number per microscopic field). Concordantly, irradiation with X-rays but not with carbon ions increased the expression of SCF and subsequently caused phosphorylation of c-Kit in endothelial cells. ISCK03 treatment of A549 cells irradiated with X-rays (6 Gy) resulted in a significant decrease in blood vessel density (X-ray, 36.44 +/- 3.44; X-ray and ISCK03, 4.33 +/- 0.71; number of microscopic field). These data indicate that irradiation of A549 cells with X rays but not with carbon ions promotes angiogenesis. CONCLUSIONS: The present study provides evidence that SCF is an X-ray-induced mediator of angiogenesis in A549 cells, a phenomenon that could not be observed with carbon ion irradiation. Thus, in this model system evaluating angiogenesis, carbon ion irradiation may have a therapeutic advantage. This observation should be confirmed in orthotopic lung tumor models. PMID- 21621346 TI - Radiotherapy plus transarterial chemoembolization for hepatocellular carcinoma invading the portal vein: long-term patient outcomes. AB - PURPOSE: We have evaluated the clinical outcomes of patients after transarterial chemoembolization (TACE) and 3-dimensional conformal radiotherapy for hepatocellular carcinoma (HCC) with portal vein tumor thrombosis (PVTT). METHODS AND MATERIALS: A registry database of 412 patients treated with TACE and three dimensional conformal radiotherapy for HCC with PVTT between August 2002 and August 2008 were analyzed retrospectively. The radiotherapy volume included the PVTT, with a 2- to 3-cm margin to cover adjacent HCC. Intrahepatic primary HCC was managed by TACE before or after radiotherapy. RESULTS: Median patient age was 52 years old, and 88.1% of patients were male. Main or bilateral PVTT was observed in 200 (48.5%) patients. Median radiation dose was 40 Gy (range, 21-60 Gy) delivered in 2- to 5-Gy fractions. We found that 3.6% of patients achieved a complete response and that 24.3% of patients achieved a partial response. The response and progression-free rates of PVTT were 39.6% and 85.6%, respectively. Median patient survival was 10.6 months, and the 1- and 2-year survival rates were 42.5% and 22.8%, respectively. Significant independent variables associated with overall survival included advanced tumor stage, alpha-fetoprotein level, degree of PVTT, and response to radiotherapy. Forty-one patients (10.0%) showed grade 3-4 hepatic toxicity during or 3 months after completion of radiotherapy. Grades 2-3 gastroduodenal complications were observed in 15 patients (3.6%). CONCLUSIONS: Radiotherapy is a safe and effective treatment for PVTT in patients with HCC. These results suggested that the combination of TACE and radiotherapy is a treatment option for relieving and/or stabilizing PVTT in patients with advanced HCC. PMID- 21621347 TI - Intragenomic and intraspecific heterogeneity of the 16S rRNA gene in seven bacterial species from the respiratory tract of cystic fibrosis patients assessed by PCR-Temporal Temperature Gel Electrophoresis. AB - 16S rRNA gene-based cultivation-independent methods are increasingly used to study the diversity of microbiota during health and disease. One bias of these methods is the variability of 16S rRNA gene that may exist among strains of a same species (intraspecific heterogeneity) or between rrs copies in a genome (intragenomic heterogeneity). We evaluated the level of intraspecific and intragenomic 16S rDNA variability in seven species frequently encountered in respiratory tract samples in cystic fibrosis (CF). A total of 179 strains were subjected to V3 region 16S rDNA PCR-TTGE. Using this easy-to-perform and rapid method, different levels of V3 region rrs heterogeneity were demonstrated. No intraspecific and intragenomic rrs heterogeneity was demonstrated for Moraxella catarrhalis (n=16), Pseudomonas aeruginosa (n=31) and Streptococcus pneumoniae (n=14) showing a single PCR-TTGE band characteristic of the species. Low level of intraspecific heterogeneity was observed for Staphylococcus aureus (n=30), Stenotrophomonas maltophilia (n=29) and Achromobacter xylosoxidans (n=28), and 17%, 38% and 96% of these strains showed intragenomic heterogeneity (two to four different rrs copies), respectively. Haemophilus influenzae (n=31) displayed the higher level of intraspecific variability with 23 different PCR-TTGE patterns and 61% of the strains showed intragenomic rrs heterogeneity (two to four different rrs copies). Although only one hypervariable region of the 16S rRNA gene was explored, intraspecific and intragenomic rrs heterogeneity was frequently observed in this study and should be taken into consideration for a better interpretation of 16S rRNA gene-based diversity profiles in denaturing gels and to avoid any overestimation of the respiratory microbiota diversity in CF. PMID- 21621348 TI - [Association between variants of lipoprotein lipase and coronary heart disease in a Tunisian population]. AB - OBJECTIVE: Coronary artery disease (CAD) is a complex multifactorial disease due to the interaction of multiple genes variations and environmental factors. Genetic variants of lipoprotein lipase (LPL), a key enzyme in the hydrolysis of triglyceride rich particles, may contribute to CAD. We analysed here the frequency of LPL variants (p.Asp9Asn, p.Asn291Ser and p.Ser447X) in a Tunisian population as well as their association with circulating lipid level and risk of CAD. PATIENTS AND METHODS: LPL variations were investigated by PCR-RFLP and lipid parameters were measured in 135 patients and 109 controls. RESULTS: The frequency of the p.Asp9Asn variation was 10.37% in CAD patients versus 3.66% in controls. The frequency for the p.Ser447X variation was 8.8% in CAD patients versus 13.7% in controls. There was no significant association between these two variants and CAD. The p.Asn291Ser mutation variation was absent in this population. In healthy subjects, heterozygote carriers of the p.Asp9Asn substitution had a significant increase level of total cholesterol (4.2+/-0.9mmol/L vs 5.6+/-1.2mmol/L; P=0.01) and a decreased level of HDL-cholesterol (1.36+/-0.3mmol/L vs 0.93+/-0.1mmol/L; P=0.045). CONCLUSION: There was no significant association between genetic variants of the LPL gene and CAD in this Tunisian population. The very low frequency of the p.Asn291Ser variation may be an ethnic specificity of Tunisians. PMID- 21621349 TI - [Serological diagnosis of celiac disease]. AB - Screening studies using high-sensitivity and specificity markers indicate a prevalence of celiac disease of up to 1% in European and North-American populations. Celiac disease is a frequent condition that has become an important public health issue. Yet the majority of cases remain undiagnosed due to the polymorphism of its clinical manifestations. The new insight in the pathogenesis of celiac disease has lead to the development of new diagnostic tools. Early screening of symptomatic patients and pre-identified at-risk groups significantly improves the quality of life while reducing morbidity and mortality. However, prophylactic benefits of early diagnosis by assessing the general population have not been shown in any study. French and Northern American scientific societies have introduced serological testing in their newly revised strategies to diagnose celiac disease. Older markers judged insufficiently accurate like anti-gliadin and anti-reticulin antibodies have recently been withdrawn from the list of reimbursed medical expenses in France. Anti-endomysium and tissue transglutaminase IgA antibodies have proven to be at this day the most sensitive and specific markers for the diagnosis and follow-up of patients on gluten-free diet, at the exception of IgA-deficient patients. Assays testing for IgG antibodies are recommended upon IgA-deficiency. Although very accurate, a better standardisation of current assays may enable serological testing to replace in a near future histological confirmation brought by small bowel biopsies which remains today the gold standard test to diagnose celiac disease. Indeed, serological testing represents and attractive alternative as it is less invasive, less expansive, laboursaving and more objective in interpretation. PMID- 21621350 TI - [Complications of celiac disease]. AB - Numerous complications can occur in celiac disease, nutritional (growth failure in children, malnutrition, vitamin deficiencies), hematologic (anaemia), bone disease (osteoporosis, fracture), gynaecologic (hypo fertility), cardiovascular (coronaropathy, venous thrombosis), neurological (peripheral neuropathy), hepatic (cytolysis, cirrhosis). Celiac disease is associated with an increased risk of autoimmune diseases (type 1 diabetes, thyroiditis), and cancer (upper digestive tract, hepatocellular carcinoma, lymphoma). The main digestive complications are microscopic colitis and refractory sprue, which are resistant to gluten-free diet. It can be associated with a monoclonal proliferation of intraepithelial lymphocytes (type 2 refractory sprue), which may be considered as a cryptic lymphoma and can lead to invasive T lymphoma, which occurs in one celiac patient in 1000. Gluten-free diet protects from the occurrence of most complications and correct the over-mortality related to these complications. PMID- 21621351 TI - Low gestational age and chronic lung disease are synergistic risk factors for retinopathy of prematurity. AB - AIMS: This retrospective, population based study was designed to investigate risk factors for development of retinopathy of prematurity (ROP) and their possible interrelationships, in neonates of gestational age (GA) <32 weeks born in a well defined geographical region. STUDY DESIGN-SUBJECTS: The study population included all preterm infants born alive with GA 24-32 weeks in Northwestern Greece during a 9-year period and hospitalised in the regional neonatal intensive care unit (NICU). OUTCOME MEASUREMENTS: The association was assessed of the presence of ROP with maternal factors: age, pathology of pregnancy, in-vitro fertilisation, multiple gestation, mode of delivery, perinatal factors: gender, antenatal steroids, transportation, resuscitation, GA, birth weight (BW), small for GA status and postnatal morbidity: chronic lung disease (CLD), intraventricular haemorrhage (IVH), necrotizing enterocolitis (NEC), respiratory distress syndrome (RDS), maximum O(2) needs, hypoxic/hyperoxic episodes, patent ductus arteriosus (PDA), sepsis, using multiple logistic regression analysis. RESULTS: Of 189 infants without congenital anomalies born at GA 24-32 weeks ROP was diagnosed in 24 (12.7%) (>grade 2: 6). Logistic regression analysis showed ROP to be strongly associated with GA, odds ratio (OR) 2.1, confidence interval (CI) 1.3-3.3, p<0.01 and CLD, OR 10.2, CI 2.3-44, p<0.01, respectively, independent of confounding factors. By estimating interaction on an additive scale it was shown that the combined risk effect of GA and CLD was larger than the sum of the individual risk effects, implying synergistic effect. CONCLUSIONS: ROP was closely and independently related to both low GA and the diagnosis of CLD, which were interrelated in the development of ROP. PMID- 21621352 TI - Cryopreservation of bull semen shipped overnight and its effect on post-thaw sperm motility, plasma membrane integrity, mitochondrial membrane potential and normal acrosomes. AB - In the Canadian Animal Genetic Resource Program, bull semen is donated in frozen or fresh (diluted) states. This study was designed to assess the cryopreservation of diluted bull semen shipped at 4 degrees C overnight, and to determine the post thaw quality of shipped semen using different straw volumes and freezing rates. Semen was collected from four breeding bulls (three ejaculates per bull). Semen was diluted in Tris-citric acid-egg yolk-glycerol (TEYG) extender, cooled to 4 degrees C and frozen as per routine (control semen). After cooling to 4 degrees C, a part of semen was removed and shipped overnight to the research laboratory via express courier (shipped semen). Semen was packaged in 0.25 or 0.5 ml straws and frozen in a programmable freezer using three freezing rates, i.e., -10, -25 or -40 degrees C/min. Control semen was also shipped to the research laboratory. Post-thaw sperm motility characteristics were assessed using CASA, and post-thaw sperm plasma membrane, mitochondrial membrane potential and normal acrosomes were assessed using flow cytometry. Post-thaw sperm quality was greater in shipped semen as compared to control (P<0.001). The shipped semen packaged in 0.25 ml straws had better post-thaw sperm quality than in 0.5 ml straws (P<0.001). Freezing rate had no effect on post-thaw sperm quality. In conclusion, bull semen can be shipped overnight for subsequent cryopreservation and gene banking. Overnight shipping of semen was found advantageous for bull semen cryopreservation. Semen packaging in 0.25 ml straws yielded better post-thaw quality than 0.5 ml straws. PMID- 21621353 TI - Calcium supplementation: balancing the cardiovascular risks. AB - Calcium supplementation has been widely accepted as a key strategy in the prevention and treatment of osteoporosis. Its role has been undermined, to some extent, by its disappointing effects on fracture in randomised controlled trials, but its use has continued to be encouraged on the grounds that it is physiologically appealing, and is unlikely to cause harm. The latter assumption is now under threat from accumulating evidence that calcium supplement use is associated with an increased risk of myocardial infarction and, possibly, stroke. The latest data, based on meta-analysis of trials involving 29,000 participants, indicate that this risk is not mitigated by co-administration of vitamin D, and that the number of cardiovascular events caused is likely to be greater than the number of fractures prevented. These findings indicate that calcium supplementation probably does not have a role as a routine preventative agent and that dietary advice is the appropriate way to attain an adequate calcium intake in most situations. Patients at high risk of fracture need to take interventions of proven anti-fracture efficacy. Available evidence suggests that this efficacy is not dependent on the co-administration of calcium supplements. PMID- 21621354 TI - Trace analysis of urea nitrate by liquid chromatography-UV/fluorescence. AB - In this paper we have adapted a technique, previously used to determine the presence of urea in aqueous samples of wine and urine, to detect trace levels of urea nitrate explosives. The procedure involves the reaction of the uronium ion (protonated urea) with a fluorophore, xanthydrol. By modification of the procedure to utilize non-aqueous reagents, in neutral conditions, it can be made specific to the presence of the urea nitrate ion pair. The procedure includes selective detection of derivatization products by UV and fluorescence following separation by High-Performance Liquid Chromatography (HPLC). Analytical method development included optimization of HPLC conditions (solvent, gradient), UV and fluorescence wavelengths, and derivatization parameters (xanthydrol amount, reaction times, temperature). The extraction of urea nitrate from surfaces was also investigated and optimized. For best quantification, it was shown that an internal standard was required; this resulted in a quantification limit around 0.17mM (21mg/L). The entire procedure could be performed in less than 30min per sample and potential interferences such as ammonium, nitrate, and urea did not produce a response under standard conditions. PMID- 21621355 TI - Determining the age at death of females in the Chinese Han population: using quantitative variables and statistical analysis from pubic bones. AB - Determining the age at death of females by skeletal features is extremely difficult and important, both in forensics and in physical anthropology. Our previous study of male pubic symphysis suggests that the indicators of morphological changes provide the best results for personal age identification. The indicators that help deduce of the age of females was approximately the same as males except for two specific indicators, which are bone density of the symphysial surface and viz. Viz are ridges and furrows on the symphysial surface, ossific nodules, lower extremities, ventral and ridge of the rampart, dorsal margin, ventral beveling and general macroscopic changes on the symphysial surface. Samples were drawn from 338 female individuals. The study procedures are as follows: Firstly, we examined the morphological features of pubic symphysis using the criteria similar to Hanihara's, Gilbert-McKern and Suchey-Brooks's methods. Secondly, we evaluated each stage with an appropriate score. Thirdly, we deduced four equations to assess the morphological features of the ages of females by statistic analysis. The results were discussed by comparing with Hanihara's, Gilbert-McKern and Suchey-Brooks's methods. The results were consistent and the developing methods for determining the age of death of females were produced. PMID- 21621356 TI - Endolaryngeal extension of thyroglossal duct cyst. AB - Thyroglossal duct cysts are the most common congenital neck masses that develop during childhood, The masses develop from remnants of thyroglossal ducts, and typically appear as midline neck masses. Endolaryngeal extension of thyroglossal duct cysts has been reported mostly as midline neck swelling. We observed a case of extension of the thyroglossal duct cyst to the supraglottic area without neck swelling. A 50-year-old man presented with a 1-month history of foreign-body sensation in the throat. Fiberscopic and radiologic findings were similar to those associated with a saccular cyst, but its proximity to the hyoid bone raised the possibility of thyroglossal duct cyst. Operation was performed via an external incision to completely remove the cyst. Postoperative fiberscopy revealed that the aryepiglottic fold swelling had disappeared. Diagnosis of thyroglossal duct cyst was confirmed on the basis of pathological findings. In cases in which it is difficult to remove the cyst from the hyoid membrane, the hyoid bone midline portion should be dissected. Thyroglossal duct cysts should be considered in cases with a submucosal tumor in the supraglottic region, and radiological examinations should be performed. PMID- 21621357 TI - Worsening of negative myoclonus by lamotrigine in a case of idiopathic focal epilepsy of children with long-term follow-up. AB - We report the case of a 5-year-old boy, first referred in 1995, with benign epilepsy with centro-temporal spikes (BECTS) with proximal negative myoclonus as the only seizure type who experienced severe aggravation of seizures when lamotrigine (25 mg/d) was prescribed in association to valproate (400 mg/d). Lamotrigine was stopped and progressively the drops of the legs disappeared within 6 months but valproate was continued. The overall prognosis was benign, confirmed by long-term follow-up until age 20, inkeeping with the diagnosis of BECTS. There was a clear relationship between introduction of lamotrigine and the detrimental effect and the condition improved dramatically when lamotrigine was stopped. Thus lamotrigine may aggravate BECTS in certain conditions and should be used with proper care in this indication. PMID- 21621358 TI - A simplified method of performance indicators development for epidemiological surveillance networks--application to the RESAPATH surveillance network. AB - BACKGROUND: Develop and calculate performance indicators allows to continuously follow the operation of an epidemiological surveillance network. This is an internal evaluation method, implemented by the coordinators in collaboration with all the actors of the network. Its purpose is to detect weak points in order to optimize management. A method for the development of performance indicators of epidemiological surveillance networks was developed in 2004 and was applied to several networks. Its implementation requires a thorough description of the network environment and all its activities to define priority indicators. Since this method is considered to be complex, our objective consisted in developing a simplified approach and applying it to an epidemiological surveillance network. METHODS: We applied the initial method to a theoretical network model to obtain a list of generic indicators that can be adapted to any surveillance network. RESULTS: We obtained a list of 25 generic performance indicators, intended to be reformulated and described according to the specificities of each network. It was used to develop performance indicators for RESAPATH, an epidemiological surveillance network of antimicrobial resistance in pathogenic bacteria of animal origin in France. CONCLUSION: This application allowed us to validate the simplified method, its value in terms of practical implementation, and its level of user acceptance. Its ease of use and speed of application compared to the initial method argue in favor of its use on broader scale. PMID- 21621359 TI - Hospital admissions related to influenza in France during the 2006/2007 epidemic. AB - BACKGROUND: The data available on hospital admissions related to influenza mostly concern in-patients admitted via the emergency department. Severe cases have been collated by intensive care practitioners since 2009. For this survey, we searched French hospital admission databases to estimate the prevalence rate of hospital admissions related to influenza and to record qualitative data. METHOD: All case studies identified between October 2006 and September 2007 were split into two groups: the first displaying symptoms of clinical influenza and the second suffering from influenza as an associated diagnosis. RESULTS: We collected 6797 hospital admissions, 2126 of which were closely related to clinical influenza. Fifty percent of cases concerned the elderly and young people. Fifty-six hospital deaths were recorded in which influenza was the underlying cause in 21% of the cases (12). When influenza was an associated diagnosis (44/56), cardiovascular or respiratory diseases were the main causes (26/44). CONCLUSION: During the same period (2006-2007), the French Sentinel Surveillance identified only 105 hospital admissions related to influenza. Our survey was therefore more exhaustive and was able to record qualitative data. Inclusion of hospital admissions with an associated diagnosis of influenza is debatable because this decreases specificity. The relationship between the principal diagnosis and all the associated diagnoses is difficult to study, although exclusion of this type of hospitalization could significantly underestimate these figures. Despite certain limitations, French hospital admissions databases should complement French Sentinel Surveillance data. PMID- 21621360 TI - Apparent diffusion coefficient values of the normal uterus: Interindividual variations during menstrual cycle. AB - OBJECTIVES: To assess the apparent diffusion coefficient (ADC) changes of the normal uterine zones among reproductive women during the menstrual cycle. METHODS: The study included 101 women of reproductive age, each with regular cycle and normal endometrium/myometrium, as proved on histopathology or MR imaging examination. Diffusion-weighted (DW) imaging was performed along the axial plane, using a single shot, multi-slice spin-echo planar diffusion pulse sequence and b-values of 0 and 800s/mm(2). The mean and standard deviation of the ADC values of normal endometrium/myometrium were calculated for menstrual, proliferative and secretory phase. Analysis of variance followed by the least significant difference test was used for statistical analysis. RESULTS: The ADC values of the endometrium were different in the three phases of the menstrual cycle (menstrual phase: 1.25+/-0.27; proliferative phase: 1.39+/-0.20; secretory phase: 1.50+/-0.18) (F: 9.64, p: 0.00). Statistical significant difference was observed among all groups (p<0.05). The ADC values of the normal myometrium were different in the three phases of the menstrual cycle (menstrual phase: 1.91+/ 0.35; proliferative phase: 1.72+/-0.27; secretory phase: 1.87+/-0.28) (F: 3.60, p: 0.03). Statistical significant difference was observed between menstrual and proliferative phase and between proliferative and secretory phase (p<0.05). No significant difference was noted between menstrual and secretory phase (p>0.05). CONCLUSIONS: A wide variation of ADC values of normal endometrium and myometrium is observed during different phases of the menstrual cycle. PMID- 21621361 TI - Pulmonary alveolar proteinosis: quantitative CT and pulmonary functional correlations. AB - OBJECTIVE: We assessed the relationship between quantitative computer tomography (qCT) and the pulmonary function test (PFT) or blood gas analysis in pulmonary alveolar proteinosis (PAP) patients, as well as the utility of these analyses to monitor responses to whole lung lavage (WLL) therapy. METHODS: Thirty-eight PAP patients simultaneously received a CT scan and PFT. Fifteen of these patients, undergoing sequential WLL for a total of 20 lavages, also underwent chest CT scans and blood gas analysis before and after WLL, and 14 of 15 patients underwent simultaneous PFT analysis. Differences between the qCT and PFT results were analyzed by canonical correlation. RESULTS: PAP patients with low predicted values for FVC, FEV1, D(LCO) and D(LCO)/VA indicated small airspace volume and mean lung inflation, low airspace volume/total lung volume ratio and high mean lung density. Correlation and regression analysis revealed a strong correlation between D(LCO) and PaO(2) values with CT results. The qCT results indicated that WLL significantly decreased lung weights and mean lung densities, and improved the total airspace volume/total lung volume ratios and mean lung inflations. CONCLUSION: Quantitative CT may be a sensitive tool for measuring the response of PAP patients to medical interventions such as WLL. PMID- 21621362 TI - Lipid emulsion as rescue therapy in lamotrigine overdose. AB - BACKGROUND: Lamotrigine is a sodium channel blocking agent that is widely prescribed for treatment of seizure. Although life-threatening effects are rarely observed in overdose, some previous reports have described the occurrence of cardiac toxicity. The management of sodium channel blocking agent-induced cardiotoxicity conventionally requires sodium bicarbonate administration. Recent case reports describe intravenous lipid administration as a successful treatment for refractory cardiovascular collapse induced by sodium channel blocking medications. OBJECTIVE: The objective of this study is to report the use of intravenous lipid emulsion as adjunctive therapy in a case of lamotrigine overdose in which electrocardiographic changes were unresponsive to bicarbonate therapy. CASE REPORT: We report a case of intentional lamotrigine overdose in a 50-year-old woman who lost consciousness and developed electrocardiographic aberrations, including widening of QRS with occurrence of left bundle branch block. The patient was initially treated with sodium bicarbonate without effect. Recovery of cardiac conduction was rapidly achieved after infusion of a 20% lipid emulsion. The exact mechanism of action of lipid emulsion is not fully understood. The lipophilic properties of lamotrigine suggest that it was partially removed by the plasmatic lipid emulsion. CONCLUSION: This case provides additional insight into the potential benefit of using lipid emulsion in refractory sodium channel blocking intoxications. PMID- 21621363 TI - Exploring strategies to improve emergency department intake. AB - BACKGROUND: The emergency department (ED) is the point of entry for nearly two thirds of patients admitted to the average United States (US) hospital. Due to unacceptable waits, 3% of patients will leave the ED without being seen by a physician. OBJECTIVES: To study intake processes and identify new strategies for improving patient intake. METHODS: A year-long learning collaborative was created to study innovations involving the intake of ED patients. The collaborative focused on the collection of successful innovations for ED intake for an "improvement competition." Using a qualitative scoring system, finalists were selected and their innovations were presented to the members of the collaborative at an Association for Health Research Quality-funded conference. RESULTS: Thirty five departments/organizations submitted abstracts for consideration involving intake innovations, and 15 were selected for presentation at the conference. The innovations were presented to ED leaders, researchers, and policymakers. Innovations were organized into three groups: physical plant changes, technological innovations, and process/flow changes. CONCLUSION: The results of the work of a learning collaborative focused on ED intake are summarized here as a qualitative review of new intake strategies. Early iterations of these new and unpublished innovations, occurring mostly in non-academic settings, are presented. PMID- 21621364 TI - Left ventricular-right atrial communication with third-degree atrioventricular block after thoracic trauma. AB - BACKGROUND: Intracardiac shunts are rare but very serious lesions after non penetrating chest trauma. Their diagnosis is difficult. This pathology often goes unrecognized in the context of multiple trauma. CASE REPORT: We report the case of a 21-year-old man involved in a motor vehicle crash who presented with multiple injuries including myocardial contusion, severe brain injury, multiple pelvic fractures, closed femur fracture, bilateral lung contusion with a right pneumothorax, and intra-abdominal injuries. Three days after the initial event, a new cardiac murmur and complete heart block appeared. Transthoracic echocardiography (echo) followed by transesophageal echo revealed a high-velocity flow communication between the left ventricle and the right atrium. The patient underwent delayed cardiac surgery due to other unstable injuries. The hospital course was prolonged but favorable, and the patient left the hospital 1 month later without any neurologic or cardiologic after-effect. CONCLUSION: This case highlights the potential for cardiac complications to occur in any patient with serious thoracic trauma. Transesophageal echo should be performed on any trauma patient with electrocardiographic abnormalities. PMID- 21621365 TI - Verbal and non-verbal behavior of doctors and patients in primary care consultations - how this relates to patient enablement. AB - OBJECTIVE: To assess the relationship between observable patient and doctor verbal and non-verbal behaviors and the degree of enablement in consultations according to the Patient Enablement Instrument (PEI) (a patient-reported consultation outcome measure). METHODS: We analyzed 88 recorded routine primary care consultations. Verbal and non-verbal communications were analyzed using the Roter Interaction Analysis System (RIAS) and the Medical Interaction Process System, respectively. Consultations were categorized as patient- or doctor centered and by whether the patient or doctor was verbally dominant using the RIAS categorizations. RESULTS: Consultations that were regarded as patient centered or verbally dominated by the patient on RIAS coding were considered enabling. Socio-emotional interchange (agreements, approvals, laughter, legitimization) was associated with enablement. These features, together with task-related behavior explain up to 33% of the variance of enablement, leaving 67% unexplained. Thus, enablement appears to include aspects beyond those expressed as observable behavior. CONCLUSION: For enablement consultations should be patient-centered and doctors should facilitate socio-emotional interchange. Observable behavior included in communication skills training probably contributes to only about a third of the factors that engender enablement in consultations. PRACTICE IMPLICATIONS: To support patient enablement in consultations, clinicians should focus on agreements, approvals and legitimization whilst attending to patient agendas. PMID- 21621366 TI - The placebo response: how words and rituals change the patient's brain. AB - OBJECTIVE: The placebo effect, or response, has evolved from being thought of as a nuisance in clinical and pharmacological research to a biological phenomenon worthy of scientific investigation in its own right. The study of the placebo effect and of its negative counterpart, the nocebo effect, is basically the study of the psychosocial context around the treatment and the patient, and it plays a crucial role in the therapeutic outcome. METHODS: In recent years, different types of placebo responses have been analyzed with sophisticated biological tools that have uncovered specific mechanisms at the anatomical, physiological, biochemical and cellular level. RESULTS: Most of our knowledge about the neurobiological mechanisms of the placebo response comes from pain and Parkinson's disease, whereby the neuronal circuits involved in placebo responsiveness have been identified. In the first case, opioidergic, dopaminergic and cholecystokinergic networks have been found to be involved. In the second case, dopaminergic activation in the striatum and neuronal changes in basal ganglia have been described. CONCLUSION: This recent research has revealed that these placebo-induced biochemical and cellular changes in a patient's brain are very similar to those induced by drugs. This new way of thinking may have profound implications both for clinical trials and for medical practice. PMID- 21621367 TI - Outcome after concomitant unilateral embolization of the internal iliac artery and contralateral external-to-internal iliac artery bypass grafting during endovascular aneurysm repair. AB - OBJECTIVE: Despite improvement of devices, endovascular aneurysm repair (EVAR) is still challenging in cases with associated aneurysmal involvement of the iliac arteries. This study examined the safety and efficacy of EVAR with concomitant unilateral embolization of the internal iliac artery (IIA) and contralateral external-to-internal iliac artery bypass grafting, with bilateral endograft limbs extended into the external iliac arteries (EIAs). METHODS: The study included 22 consecutive patients (mean age, 74 years) who underwent elective endovascular repair of aortoiliac or iliac aneurysms, with concomitant coil embolization of the unilateral IIA and contralateral EIA-to-IIA bypass in the same operative setting. Five patients had a unilateral IIA aneurysm, and eight had bilateral IIA aneurysms. EIA-to-IIA bypass grafting was performed through the retroperitoneal approach. The perioperative and midterm outcome of the procedure was assessed. RESULTS: The procedure was successfully performed in all cases. Eleven patients underwent IIA embolization at the main trunk, and the other 11 cases required IIA occlusion at distal branches. There was no perioperative death or severe complication. The mean follow-up period was 15.7 +/- 7.8 months, ranging from 2 to 32 months. The bypass remained patent in all cases, and there was no occurrence of graft-related complication. Enlargement of aneurysms or development of type I endoleak was not observed. Persistent mild buttock claudication occurred in two patients (9%) ipsilaterally to the occluded IIA; one patient after IIA occlusion at the main trunk and the other at distal branches. No other pelvic ischemic manifestation was observed. CONCLUSIONS: EVAR with simultaneous unilateral IIA embolization and contralateral EIA-to-IIA bypass grafting is feasible, with a relatively low risk of complications. It can be a useful treatment option in cases with complex aortoiliac aneurysms, including those with bilateral IIA aneurysms. PMID- 21621368 TI - Proceedings of the 2009 annual meeting of the Fetal Alcohol Spectrum Disorders Study Group. AB - The annual meeting of the Fetal Alcohol Spectrum Disorders Study Group (FASDSG) was held on June 20, 2009 in San Diego, CA, as a satellite of the Research Society on Alcoholism Meeting. The FASDSG membership includes clinical, basic, and social scientists who meet to discuss recent advances and issues in Fetal Alcohol Spectrum Disorders research. The main theme of the meeting was "Epigenetics and Development." Two keynote speakers, Dr. Randy Jirtle and Dr. Michael Skinner, addressed the role of epigenetics and environmental inputs, including alcohol, during critical stages of development and their potential critical and long-lasting effects. Members of the FASDSG provided new findings through brief "FASt" data reports, and national agency representatives provided updates on activities and funding priorities. Scientific presentations were made by recipients of the Student Research Merit Award and Rosett Award. PMID- 21621369 TI - Associations of corticosterone and testosterone with alcohol drinking in F2 populations derived from AA and ANA rat lines. AB - In our previous studies on alcohol-preferring AA (Alko alcohol) and nonpreferring ANA (Alko nonalcohol) rats, we have observed that the AA rats exhibit lower endogenous levels of corticosterone, higher testosterone levels, and more frequent alcohol-induced testosterone elevations when compared with ANA rats. The objective of the present study was to get more conclusive evidence for the potential role of the hypothalamus-pituitary-adrenal and hypothalamus-pituitary gonadal axes in alcohol drinking by using the F2 experimental design. Alcohol preferring AA and alcohol-nonpreferring ANA rat lines were crossbred to form a F1 population from which the final F2 population was derived. Male animals were challenged with a priming alcohol dose after which a 3 weeks' voluntary alcohol drinking period took place. After a washout period of 1 week, one-half of the 40 highest and 40 lowest alcohol drinkers were challenged with a second dose of alcohol and the other half with saline. Serum testosterone and corticosterone levels were measured before and during the test. Higher endogenous testosterone levels were detected in the rats of the high alcohol consumption group compared with the low consumption group. Also supporting the original AA/ANA line differences, a trend for lower endogenous corticosterone levels were measured in the high alcohol consumption group compared with the low consumption group. The alcohol challenge test after the drinking period resulted in a higher frequency (38%) of testosterone elevations in the high drinkers compared with the low drinkers (5%). The present data confirms the validity of the positive connections between testosterone elevation and increased alcohol drinking, as well as between testosterone reduction and decreased alcohol drinking, in AA and ANA rats. PMID- 21621370 TI - Orexin A expression and promoter methylation in patients with alcohol dependence comparing acute and protracted withdrawal. AB - The orexins (hypocretins) are neuropeptides deriving from the lateral hypothalamus and may be of importance within the context of drug craving, withdrawal, and relapse. Therefore, the orexin A expression and promoter methylation in peripheral blood cells of 68 patients (41 male and 27 female patients at three different time points during withdrawal and 27 patients during stationary dehabituation therapy) suffering from alcohol dependence were assessed by quantitative reverse transcription-polymerase chain reaction and bisulfite sequencing. There was a statistically significant difference of orexin A expression between the three time points of withdrawal and long-term (LT) abstinence (F=4.16, P=.011). This difference was most prominent in comparison with LT abstinence (t=-3.08, P=.0032). Expression was significantly associated with the severity of withdrawal symptoms measured with the Withdrawal Syndrome Scale for Alcohol and Related Psychoactive Drugs (WSA) (t=2.17, P=.0356). The stronger the withdrawal symptoms, the lower the orexin A expression (F=4.69, P=.036). Body mass index (t=2.15, P=.041), the severity of withdrawal measured with the WSA (t=2.595, P=.0133), craving measured either by the Obsessive Compulsive Drinking Scale (t=2.77, P=.0085) or the Lubecker Craving Questionnaire (t=-2.23, P=.0314) had a significant influence on orexin A expression taking into account mean methylation of the CpG island of the orexin A promoter during withdrawal. Orexin A may be a possible candidate to further elucidate mechanisms of alcohol withdrawal taking into account energy homoeostasis in the circuit of reward and motivation. PMID- 21621372 TI - Relation of tunnel enlargement and tunnel placement after single-bundle anterior cruciate ligament reconstruction. AB - PURPOSE: To determine the relation between tunnel placement and tunnel enlargement after single-bundle anterior cruciate ligament (ACL) reconstruction. METHODS: Seventy-two subjects (mean age, 30.1 years; 17.5 months' follow-up) who underwent single-bundle ACL reconstruction with hamstring autograft were studied. EndoButton fixation (Ethicon, Somerville, NJ) was used on the femoral side, whereas staples were used on the tibial side. A transtibial femoral tunnel position technique was used in 53 subjects, whereas positioning through the medial portal was used in the other 19 cases. Tunnel enlargement was determined by comparing the diameter of the tunnel on the radiograph obtained after 12 months and the radiograph obtained instantly after the operation. The centers of the femoral and tibial tunnels and the angles between the graft and tibial plateau were also measured on standard radiographs. Clinical outcomes including KT-1000 assessment (MEDmetric, San Diego, CA) and International Knee Documentation Committee (IKDC) score were also collected in all patients. The relations between tunnel enlargement/tunnel position and knee joint laxity and IKDC score were analyzed. RESULTS: The mean KT-1000 side-to-side difference significantly decreased, from 6.07 +/- 2.75 mm to 1.57 +/- 2.14 mm, after ACL reconstruction; the IKDC subjective score increased from 52.8 to 87.5. On lateral radiographs, the tunnel enlargement rates were 41% on the femoral side and 35% on the tibial side; on plain anteroposterior radiographs, the tunnel enlargement rates were 39% on the femoral side and 32% on the tibial side. Subjects with a higher femoral tunnel had a greater enlargement rate (P < .001). Subjects with a more vertical graft also had a larger femoral enlargement (P < .05). More anterior placement of the femoral tunnel was associated with larger tibial tunnel enlargement on anteroposterior plain radiographs (P < .05). A more vertical graft was also associated with larger tibial tunnel enlargement. Subjects in whom the transtibial femoral position technique was used had more femoral tunnel enlargements (P < .01). CONCLUSIONS: Drilling the femoral tunnel through the medial portal created a lower, more posterior, and less vertical tunnel than drilling through the tibial tunnel. Femoral and tibial tunnel enlargements were greater with more anterior, more proximal, and more vertical femoral tunnels. Whereas no clinical differences were seen in the 2 groups, drilling the femoral tunnel from the medial portal will result in smaller postoperative tunnel enlargements. LEVEL OF EVIDENCE: Level III, retrospective comparative study. PMID- 21621371 TI - Effects of alcohol intoxication and gender on cerebral perfusion: an arterial spin labeling study. AB - An increasing number of studies use functional MRI (fMRI) and blood oxygen level dependent (BOLD) signal to investigate the neurofunctional basis of acute alcohol effects on the brain. However, the BOLD signal reflects neural activity only indirectly as it depends on regional hemodynamic changes and is therefore sensitive to vasoactive substances, such as alcohol. We used MRI-based pulsed arterial spin labeling (ASL) method to quantify effects of acute intoxication on resting cerebral perfusion. Gender effects have not been previously examined and yet they are of particular interest given the differences in hormonal dynamics, alcohol metabolism, and hemodynamic regulation. Nineteen young, healthy individuals (nine women) with no personal or familial alcohol- or drug-related problems served as their own controls by participating in both alcohol (0.6g/kg ethanol for men, 0.55g/kg for women) and placebo scanning sessions in a counterbalanced manner. Regionally specific effects of the moderate alcohol dose on gray matter perfusion were examined with voxel-wise and region-of-interest analyses suggesting an interaction between gender and alcohol beverage. Acute intoxication increased perfusion in bilateral frontal regions in men but not in women. Under placebo, stronger cortical perfusion was observed in women compared with men primarily in the left hemisphere in frontal, parietal, and temporal areas. These results emphasize gender differences and regional specificity of alcohol's effects of cerebral perfusion possibly because of interactive influences on hormonal, metabolic, and hemodynamic autoregulatory systems. Alcohol-induced perfusion increase correlated positively with impulsivity/antisocial tendencies, consistent with dopaminergic mediation of reward, and its effects on cortical perfusion. Additional ASL studies are needed to investigate dose- and time-dependent effects of alcohol intoxication and gender on the hemodynamic factors that conjointly influence BOLD signal to disambiguate the vascular/metabolic mechanisms from the neurally based changes. PMID- 21621373 TI - Outcome after partial medial meniscus substitution with the collagen meniscal implant at a minimum of 10 years' follow-up. AB - PURPOSE: The aim of the study was to evaluate the clinical outcome of a collagen meniscus graft implanted in an injured medial meniscus after a minimum of 10 years' follow-up. METHODS: Twenty-five patients underwent arthroscopic implantation of the collagen meniscus device. They had either persistent compartmental joint line pain due to a previous medial meniscus resection (5 cases) or a large irreparable meniscus tear at arthroscopy (20 cases). Implant failure was defined as infection due to the implant or mechanical failure of the device. Twenty-two patients returned for clinical, functional, and radiographic evaluation. Magnetic resonance imaging was also performed and was analyzed with the criteria of Genovese et al. (where type 3 indicates normal and type 1 indicates completely abnormal). All the aforementioned evaluations were carried out at a minimum of 10 years (range, 10.1 to 12.5 years) after the procedure. RESULTS: The mean Lysholm score improved from 59.9 preoperatively to 89.6 at 1 year (P < .001), and it was 87.5 at final follow-up (P < .001). The results were good or excellent in 83% of the population. No differences were observed when we compared the Lysholm score at 1 year of follow-up with the score at final follow up (P > .05). The mean pain score on a visual analog scale improved by 3.5 points at final follow-up. Patient satisfaction with the procedure was 3.4 of 4 points. Radiographic evaluation showed either minimal or no narrowing of the joint line. Magnetic resonance imaging showed type 2 in 64% of cases and type 3 in 21%. All cases showed less volume than expected (size type 2 in 89%). The failure rate in the patient population was 8% (2 of 25). There were no complications related to the device. CONCLUSIONS: Although there were several different types of patients and acute and chronic tears were treated in a limited number of patients, meniscal substitution with the collagen meniscal implant provides significant pain relief and functional improvement after a minimum of 10 years' follow-up. The implant generally diminished in size, but the procedure proved to be safe and had a low rate of implant failure on a long-term basis. No development or progression of degenerative knee joint disease was observed in most cases. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 21621374 TI - Short- and long-term outcomes in children undergoing radiofrequency catheter ablation before their second birthday. AB - BACKGROUND: Tachyarrhythmias can cause substantial morbidity and mortality in infants and very young children. Our objective was to assess early and late outcomes in children undergoing radiofrequency catheter ablation before their second birthday. METHODS: This is a retrospective review of medical records from 1995 till 2009 at a single institution with a large referral base. RESULTS: Thirty-four children younger than 2 years of age were brought to the electrophysiology laboratory for catheter ablation for tachyarrhythmia. Radiofrequency ablation (42 procedures) was performed in 31 children (mean weight, 7.4 kg; range, 2.6-12.3 kg). Tachyarrhythmias were atrioventricular re entry tachycardia (19), ectopic or multifocal atrial tachycardia (6), atrial flutter (1), ventricular tachycardia (3), and congenital junctional ectopic tachycardia (2). Seventeen children presented with tachycardia-induced cardiomyopathy and heart failure. Three patients required extracorporeal life support prior to the procedure. Radiofrequency catheter ablation was successful in 74% of these very young children vs 91% in a comparison group of children older than 2 years (n = 447). Among patients with initially successful ablations, there was no tachyarrhythmia recurrence during medium- to long-term follow-up (1.4-15.0 years; mean, 7.3 years). Major complications occurred in 4 very young children. Eight of 11 patients in whom ablation failed or could not be done required another form of continued aggressive treatment. CONCLUSION: Catheter ablation is indicated and successful in critically ill infants with life threatening tachyarrhythmia refractory to medical therapy. Initially successful procedures are associated with excellent long-term outcomes, but significant complication rates require that such procedures be reserved for carefully selected patients and performed by highly skilled staff. PMID- 21621375 TI - A novel nonsense variant in Nav1.5 cofactor MOG1 eliminates its sodium current increasing effect and may increase the risk of arrhythmias. AB - BACKGROUND: The protein MOG1 is a cofactor of the cardiac sodium channel, Nav1.5. Overexpression of MOG1 in Nav1.5-expressing cells increases sodium current markedly. Mutations in the genes encoding Nav1.5 and its accessory proteins have been associated with cardiac arrhythmias of significant clinical impact. We sought to investigate whether MOG1 is implicated in cardiac arrhythmias. METHODS: We performed a genetic screening of the MOG1-encoding gene (gene symbol RANGRF, alias MOG1) in 220 Danish patients with cardiac arrhythmia. Of the 220, 197 were young patients with lone atrial fibrillation and 23 were patients with Brugada syndrome. The effect of one variant was investigated functionally by patch clamping CHO-K1 cells coexpressing Nav1.5 with MOG1. RESULTS: We uncovered a novel heterozygous nonsense variant, c.181G>T (p.E61X), that, however, was also present in control subjects, albeit at a lower frequency (1.8% vs 0.4%, P = 0.078). Electrophysiological investigation showed that the p.E61X variant completely eliminates the sodium current-increasing effect of MOG1 and thereby causes loss of function in the sodium current. When mimicking heterozygosity by coexpression of Nav1.5 with wild-type MOG1 and p.E61X-MOG1, no current decrease was seen. CONCLUSIONS: Our screening of Nav1.5 cofactor MOG1 uncovered a novel nonsense variant that appeared to be present at a higher frequency among patients than control subjects. This variant causes MOG1 loss of function and therefore might be disease causing or modifying under certain conditions. PMID- 21621376 TI - Endovascular repair of a profunda femoris artery aneurysm. AB - Profunda femoris artery (PFA) aneurysms are rare and they are usually treated with open surgery. We report the case of an 87-year-old patient who during the preoperative work-up for an abdominal aortic aneurysm was found to have a 4.2-cm aneurysm of the right PFA. A percutaneous transcatheter coil embolization was successfully performed using a contralateral retrograde approach. To date, only three patients, including the present case, have been treated with transcatheter coil embolization for a PFA aneurysm. In the absence of concomitant superficial femoral artery disease, embolization of aneurysms of the profunda femoris seems to be a safe and minimally invasive alternative to open repair in selected patients. PMID- 21621377 TI - Complications of the access during aortic valve implantation through transfemoral access. AB - BACKGROUND: Aortic valve implantation (AVI) is a booming therapeutic option in high-risk patients with calcific aortic stenosis. Retrograde femoral approach drawbacks include vascular complications owing to the size of the introduction system (22- and 24-F).The aim of this study was to retrospectively analyze the incidence and the treatment of vascular complications in the first 2 years of transfemoral AVI experience with the first generation of Edwards SAPIEN transcatheter heart valves. METHODS: Since December 2007, AVI has been performed in 71 patients, 21 times by the transapical route and 50 times by the transfemoral route through an inguinal approach with the first generation of Edwards SAPIEN transcatheter heart valves (23 and 26 mm). The incidence and the treatment of vascular complications were evaluated as main criteria for transfemoral AVI. RESULTS: All the procedures could be successfully performed by a femoral route, except for three cases when the introducing device could not be fixed on the thoracic aorta because of vascular access problems. Vascular access related complications occurred in nine patients (18%), including three iliac dissections, two aortic dissections, three femoral lesions, and one thoracic aorta rupture. These complications were treated either in a conservative way (n = 2), or in an endovascular way using a contralateral approach (n = 3), or surgically through an inguinal approach (n = 3). A traumatic rupture of the thoracic aorta resulted in the death of a female patient. CONCLUSION: In our experience, transfemoral AVI gives a satisfying technical success rate in the selected patients. The incidence of complications involving the vascular access remains an important limitation of this new technique. Although a conservative or endovascular treatment can be applied in most cases, improving the introduction devices is highly expected because it would reduce the complications rate of vascular access. PMID- 21621378 TI - Endovascular management of type B aortic dissection after attempted central venous catheterization. AB - A 41-year-old woman, status postmastectomy for breast cancer had an attempted 8-F left subclavian vein chemotherapy port placed in her. She developed severe upper back pain radiating to the left shoulder. A computed tomographic scan and angiography revealed catheter placement in the left subclavian artery and a type B aortic dissection. A thoracic stent-graft was used to treat the aortic dissection. While removing the catheter, a covered stent was deployed to seal the arterial puncture and a balloon-expandable stent was placed over a persistent subclavian dissection. This case illustrates an example of the feasibility of endovascular management to treat serious iatrogenic access complications. PMID- 21621379 TI - Open and endovascular management of concomitant severe carotid and coronary artery disease: tabular review of the literature. PMID- 21621380 TI - Acoustical study of classical Peking Opera singing. AB - Acoustic characteristics of classical opera singing differ considerably between the Western and the Chinese cultures. Singers in the classical Peking opera tradition specialize on one out of a limited number of standard roles. Audio and electroglottograph signals were recorded for four performers of the Old Man role and three performers of the Colorful Face role. Recordings were made of the singers' speech and when they sang recitatives and songs from their roles. Sound pressure level, fundamental frequency, and spectrum characteristics were analyzed. Histograms showing the distribution of fundamental frequency showed marked peaks for the songs, suggesting a scale tone structure. Some of the intervals between these peaks were similar to those used in Western music. Vibrato rate was about 3.5Hz, that is, considerably slower than in Western classical singing. Spectra of vibrato-free tones contained unbroken series of harmonic partials sometimes reaching up to 17 000Hz. Long-term-average spectrum (LTAS) curves showed no trace of a singer's formant cluster. However, the Colorful Face role singers' LTAS showed a marked peak near 3300Hz, somewhat similar to that found in Western pop music singers. The mean LTAS spectrum slope between 700 and 6000Hz decreased by about 0.2dB/octave per dB of equivalent sound level. PMID- 21621381 TI - Frame by frame analysis of glottic insufficiency using laryngovideostroboscopy. AB - OBJECTIVES/HYPOTHESIS: Glottic insufficiency (GI) can be either grossly obvious or subtle in its presentation. Subtle GI is demonstrated by various Laryngovideostroboscopic (LVS) clues, including complete but "short" phase closure of the true vocal folds (VFs) during the glottic cycle. We used the frame by frame analysis (FBFA) technique to evaluate its effectiveness in objectively contributing to the diagnosis of subtle GI in patients with atrophic and/or paretic VFs. This article intends to formally present the methods and intentions of the FBFA technique and report our findings using FBFA on subjects with clinically diagnosed GI and normal volunteers. STUDY DESIGN: Retrospective review and demonstration of technique. METHODS: Forty-four subjects with a prior clinical diagnosis of true VF atrophy (25/44) and/or paresis (19/44) and five normal volunteers were identified. Using the FBFA technique, each subject's average percentage of closed frames per glottic cycle was recorded. RESULTS: Subjects with atrophy spent 32.4% of the frames of the glottic cycle in the closed phase, subjects with paresis spent 35.7% of the frames closed, and normal subjects spent 50.2% of the frames closed. CONCLUSIONS: FBFA appears to be a simple objective method for the novice or experienced LVS interpreter, by which one can suspect subtle GI. Because of the inherent physical properties by which LVS gives an "illusionary" representation of the glottic cycle, the FBFA technique remains a theoretical tool. Future studies using high-speed digital imaging are needed to validate this useful technique. PMID- 21621382 TI - Acoustic and perceptual aspects of vocal function in children with adenotonsillar hypertrophy--effects of surgery. AB - OBJECTIVE: To evaluate outcome of two types of tonsil surgery (tonsillectomy [TE]+adenoidectomy or tonsillotomy [TT]+adenoidectomy) on vocal function perceptually and acoustically. STUDY DESIGN: Sixty-seven children, aged 50-65 months, on waiting list for tonsil surgery were randomized to TE (n=33) or TT (n=34). Fifty-seven age- and gender-matched healthy preschool children were controls. Twenty-eight of them, aged 48-59 months, served as control group before surgery, and 29, aged 60-71 months, served as control group after surgery. METHODS: Before surgery and 6 months postoperatively, the children were recorded producing three sustained vowels (/alpha/, /u/, and /i/) and 14 words. The control groups were recorded only once. Three trained speech and language pathologists performed the perceptual analysis using visual analog scale for eight voice quality parameters. Acoustic analysis from sustained vowels included average fundamental frequency, jitter percent, shimmer percent, noise-to-harmonic ratio, and the center frequencies of formants 1-3. RESULTS: Before surgery, the children were rated to have more hyponasality and compressed/throaty voice (P<0.05) and lower mean pitch (P<0.01) in comparison to the control group. They also had higher perturbation measures and lower frequencies of the second and third formants. After surgery, there were no differences perceptually. Perturbation measures decreased but were still higher compared with those of control group (P<0.05). Differences in formant frequencies for /i/ and /u/ remained. No differences were found between the two surgical methods. CONCLUSION: Voice quality is affected perceptually and acoustically by adenotonsillar hypertrophy. After surgery, the voice is perceptually normalized but acoustic differences remain. Outcome was equal for both surgical methods. PMID- 21621383 TI - The vocal load of Reform Jewish cantors in the USA. AB - OBJECTIVE: Jewish cantors comprise a subset of vocal professionals that is not well understood by vocal health professionals. This study aimed to document the vocal demands, vocal training, reported incidence of voice problems, and treatment-seeking behavior of Reform Jewish cantors. METHODS: The study used a prospective observational design to anonymously query Reform Jewish cantors using a 35-item multiple-choice survey distributed online. Demographic information, medical history, vocal music training, cantorial duties, history of voice problems, and treatment-seeking behavior were addressed. RESULTS: Results indicated that many of the commonly associated risk factors for developing voice disorders were present in this population, including high vocal demands, reduced vocal downtime, allergies, and acid reflux. Greater than 65% of the respondents reported having had a voice problem that interfered with their ability to perform their duties at some time during their careers. CONCLUSION: Reform Jewish cantors are a population of occupational voice users who may be currently unidentified and underserved by vocal health professionals. The results of the survey suggest that Reform Jewish cantors are occupational voice users and are at high risk for developing voice disorders. PMID- 21621384 TI - Application of texture analysis to ventilation SPECT/CT data. AB - It is demonstrated that textural parameters calculated from functional pulmonary CT data have the potential to provide a robust and objective quantitative characterisation of inhomogeneity in lung function and classification of lung diseases in routine clinical applications. Clear recommendations are made for optimum data preparation and textural parameter selection. A new set of platform independent software tools are presented that are implemented as plug-ins for ImageJ. The tools allow segmentation and subsequent histogram-based and grey level co-occurrence matrix based analysis of the regions of interest. The work flow is optimised for use in a clinical environment for the analysis of transverse Computed Tomography (CT) scans and lung ventilation scans based on SPECT. Consistency tests are made against other texture analysis plug-ins and simulated lung CT data. The same methods are then applied to patient data consisting of a healthy reference group and one patient group each who suffered from asthma, chronic obstructive pulmonary disease (COPD), and COPD plus lung cancer. The potential for disease classification based on computer analysis is evaluated. PMID- 21621385 TI - Chella David: a lifetime contribution in translational immunology. AB - Immunology, like most scientific disciplines, is filled with buzz words. One such buzz word or term has been coined "translational immunology". Indeed, translational research is amongst the most popular expressions used to justify the use of basic research in the hopes that such research will lead to solutions to clinical problems. In fact, no such justification is needed and some of the most important observations in clinical medicine have been derived from basic science; basic science that had no idea at its time of its usefulness in clinical medicine. This special issue is devoted to Chella David. Chella's contributions in immunology have been legion. Before inbred mice became popular, long before multi-million dollar companies were developed to develop such models, Chella David was hard at the bench studying the genetics of the murine immune system and the importance of such mouse models in autoimmune diseases. Importantly, Dr. David provided animals without strings, without the burdens of MTAs, that now impede research. Chella has been generous with his time, with his reagents, and has been a caring and devoted mentor to generations of students. This issue is part of our series to recognize autoimmunologists and dedicated themes that include papers in multiple disciplines of immunology, but especially are focused on cutting-edge applications that will improve clinical therapeutics. Chella David, at age 75, is an athlete in immunology and still keeps going with the same enthusiasm as manifest as a young post-doc. PMID- 21621386 TI - Hypolipidemic effect of mustard oil enriched with medium chain fatty acid and polyunsaturated fatty acid. AB - OBJECTIVE: The hypolipidemic effect of two structured lipids prepared from mustard oil was studied: one was medium chain fatty acid (MCFA) rich mustard oil and the other was polyunsaturated fatty acid (PUFA) rich mustard oil. METHODS: Thirty-six rats were involved in the study with six rats in each group. Three groups were normal, whereas the others were hypercholesterolemic. Hypolipidemic effects were investigated in both plasma and tissues. RESULTS: Both structured lipids with similar content of functional fatty acids (about 19%) were fed (20% of diet weight) to normal and hypercholesterolemic male albino rats. After 28 d of feeding, the fat and protein digestibility increased by administration of the two structured lipids in comparison with the control group fed with normal mustard oil. In both normal and hypercholesterolemic condition plasma cholesterol, non-high-density lipoprotein, and triacylglycerols were reduced by feeding the rats with experimental oils, but the reduction was in lesser amount in rats fed with MCFA-rich mustard oil than in rats fed with PUFA-rich mustard oil and same findings were also seen in hypercholesterolemia. However the HDL levels increased with the administration of both experimental oils. CONCLUSION: Mustard oil can be enriched with MCFA and n-3 PUFA and these modified lipids exhibited hypocholesterolemic and hypolipidemic effects in rats when compared to unmodified mustard oil. PMID- 21621387 TI - Nutritional support in the treatment of aplastic anemia. AB - OBJECTIVE: Whether a specific nutritional support promotes healing of aplastic anemia (AA) patients is still unclear. Therefore, we explored the potential of a high-nucleotide, arginine, and micronutrient nutritional supplement on the nutritional rehabilitation of AA mice. METHODS: The BALB/c AA mice model was treated with hypodermic injections of acetylphenylhydrazine (100 mg/kg), x-ray (2.0 Gy), and intraperitoneal injections of a cyclophosphamide (80 mg/kg) combination. Then AA mice were fed with nutritional supplements in a dose dependent manner (1445.55, 963.7, 674.59 mg/kg/d) for 7 wk. At the end of the experimental period, mice were autopsied. A full blood count was performed, and femoral marrow cell suspensions were prepared to assess the total femoral nucleated cell count and the number of committed hemopoietic progenitor cells (colony-forming units). The pathologic changes of liver and spleen were analyzed. RESULTS: The significant increases of nutrient mixture groups were evident in many peripheral blood parameters. The femoral nucleated cell count and colony forming units of nutritional supplements groups were markedly increased, compared with the AA group. Transmission electron microscopy showed that the number of mitochondria in similar bone marrow cells was increased in nutritional supplements groups. The nutritional supplements also affected the recovery of livers and spleens of AA mice. CONCLUSION: Specific nutritional supplements accelerated rehabilitation of AA mice and can be used as nutritional support in the treatment of AA. PMID- 21621388 TI - Metabolic syndrome after liver transplantation: prevalence and predictive factors. AB - OBJECTIVES: Metabolic syndrome (MetS) is a disorder in which obesity, insulin resistance, high blood pressure and dyslipidemia coexist. This study assessed the prevalence of MetS and its associated factors in patients who underwent orthotopic liver transplantation (OLTx). METHODS: Post-OLTx patients were assessed for the presence of MetS according to the diagnostic criteria proposed by the International Diabetes Federation (IDF) and National Heart, Lung, and Blood Institute/American Heart Association (NHLBI/AHA). Demographic, socioeconomic, lifestyle, clinical, anthropometric, and dietary variables were collected to identify predictors for MetS using logistic regression analysis. RESULTS: Among the 148 patients assessed, the prevalence of MetS was 50% (IDF criteria) and 38.5% (NHLBI/AHA criteria). For both the IDF and the NHLBI/AHA classifications, the independent factors associated with MetS were older age, shorter time since transplantation, and history of excessive weight prior to OLTx. Other predictors for MetS by IDF criteria were alcohol abuse as the indication for OLTx, physical activity reduction as the cause of weight gain after transplantation, and calcium intake below recommended levels. The presence of MetS (NHLBI/AHA) was also associated with decreased intake of potassium, fiber, and folic acid. CONCLUSIONS: MetS is highly prevalent among post-OLTx patients and it is predicted by older age, shorter time since transplantation, alcohol abuse as the cause of cirrhosis, excessive weight prior to OLTx, and some potentially modifiable factors such as physical activity reduction after OLTx and low intake of calcium, potassium, fiber, and folic acid. PMID- 21621389 TI - Iodine nutritional status in pregnant women of two historically different iodine deficient areas of Catalonia, Spain. AB - OBJECTIVE: Catalonia (Spain) has a historically worse situation of mild iodine deficiency in the Pyrenees Mountains compared with the coastal region. The aim of this study was to evaluate the current iodine status in pregnant women living in these two areas. METHODS: An epidemiologic prospective survey included 267 consecutive pregnancies in the Catalan mountains (n = 139) and coast (n = 128) studied during the first trimester; an additional subset of 135 women from the initial cohort was available for evaluation in the third trimester. Urinary iodine (UI) was measured, and questionnaires to determine iodized salt and sea fish consumption and potassium iodide supplementation were administered. RESULTS: The median UI in the first trimester was 163 MUg/L for the entire cohort, with differences between mountain and coastal regions (209 versus 142 MUg/L, P = 0.007). The highest prevalence of iodized salt consumption was in the mountain area (58% versus 36.4%, P < 0.001). For the entire group, a higher median UI was found in iodized salt consumers compared with non-consumers (193 versus 134 MUg/L, P < 0.001). In the third trimester, an increase of median UI was seen in those to whom iodine supplements were given during pregnancy (190 versus 154 MUg/L, P = 0.015). CONCLUSION: A reversal in the historically iodine-deficient situation was observed in the Catalan Pyrenees compared with the coastal area, with a globally acceptable iodine status in pregnant women of the two geographic locations. Iodized salt consumption seems to have contributed to maintaining an acceptable iodine status in this population. PMID- 21621390 TI - Successful treatment of an infected wound in infants by a combination of negative pressure wound therapy and arginine supplementation. AB - OBJECTIVE: Wound dehiscence caused by surgical site infection (SSI) presents a complicated problem. Negative pressure wound therapy (NPWT) was developed to treat wound dehiscence. Nutritional treatment using arginine has also been recently shown to be effective for the treatment of pressure ulcers. Therefore, wound complications due to SSI were treated using NPWT combined with nutritional therapy with an arginine-rich supplement (ARS). METHODS: Six pediatric patients with wound dehiscence due to SSI received this combined therapy. RESULTS: The average age of the patients was 12.2 mo. The operations that these patients underwent included laryngotracheal separation, radical operation for spinal bifida, gastrostomy, colostomy, anorectoplasty, and tumor extirpation. A local wound infection induced wound dehiscence in all patients. Therefore, NPWT was introduced with an enteral administration of ARS. All wounds completely healed within 1 mo after the introduction of this combined therapy without any other complications from the NPWT or ARS. A follow-up study at 6 mo after this therapy was completed showed no complications associated with the wounds. CONCLUSION: This combination therapy using NPWT and ARS administration was effective in inducing early healing of infected wound complications after surgery. PMID- 21621391 TI - Freeze-dried instant coffee can promote the activities of antioxidant enzymes and induce weight loss but also aggravate the plasma cholesterol profile in rats. AB - OBJECTIVE: This study investigated the effect of instant coffee on antioxidant enzyme activity and plasma cholesterol profile during exercise in rats. METHODS: Forty eight rats were fed a control diet with water (C) or a control diet with a coffee solution (CF). At the end of week 4, animals in each dietary group were subdivided into three exercise groups: before exercise (BE), during exercise (DE), and after exercise (AE). DE groups were exercised on a treadmill for 1 h immediately before being sacrificed. Animals in the AE groups were allowed to take a rest for 1 h after exercise. Antioxidant enzyme activities of the C and CF groups were evaluated with activities of catalase in plasma and superoxide dismutase, the ratio of reduced glutathione to oxidized glutathione, and the level of malondialdehyde in the liver. Plasma concentrations of triacylglycerol, total cholesterol, and high-density lipoprotein cholesterol were also compared. RESULTS: Final body weights and food intakes of the CF group were significantly lower than those of the C group. Catalase activities of the CF group were higher than those of the C group BE and AE. Reduced glutathione/oxidized glutathione of the CF group was significantly higher than that of the C group BE and DE. Superoxide dismutase activities of the CF group were higher than those of the C group regardless of exercise. Compared with the C group, there was an increase of total cholesterol and a decrease of high-density lipoprotein cholesterol levels in the CF group. Malondialdehyde levels in the CF group were higher than those in the C group BE and AE. CONCLUSION: It is suggested that freeze-dried instant coffee can promote activities of antioxidant enzymes and induce weight loss but also aggravate the plasma cholesterol profile in rats. PMID- 21621392 TI - Total and abdominal obesity among rural Chinese women and the association with hypertension. AB - OBJECTIVE: Obesity increases the risk of hypertension and other chronic diseases, which are little known in rural China. This study aimed to investigate the epidemiologic features and the association with hypertension of obesity in rural Chinese women. METHODS: A cross-sectional survey was conducted during 2004 through 2006, which used a multistage cluster sampling method to select a representative sample in Liaoning Province, China. In total 23 178 rural participants at least 35 y of age were examined (the percentage of subjects >64 y old was 14.5%). Data on demographic variables (age, sex, and race), smoking status, use of alcohol, physical activity, and education level were obtained by interview. Overweight and obesity were defined according to the World Health Organization classification. Hypertension was defined according to the criteria established by the Seventh Report of the Joint National Committee, and untreated hypertensive subjects were further classified into three subtypes: isolated systolic hypertension, isolated diastolic hypertension, and systolic and diastolic hypertension. Multivariable models and performed Poisson logistic regression analysis were used to determine associations among body mass index (BMI), waist circumference, and variables. RESULTS: Overall, the prevalences of overweight and obesity were 24.4% and 2.7%, respectively, as defined by BMI, whereas the prevalences were 48.6% and 4.9% as defined by waist circumference. Poisson regression revealed that high levels of physical activity (defined by BMI, moderate: prevalence ratio [PR] 0.976, 95% confidence interval [CI] 0.965 0.988, high: PR 0.985, 95% CI 0.971-0.999; defined by waist circumference, moderate: PR 0.955, 95% CI 0.944-0.965, high: PR 0.973, 95% CI 0.960-0.985) and current smoking status (defined by BMI, PR 0.950, 95% CI 0.938-0.962; defined by waist circumference, PR 0.966, 95% CI 0.954-0.978) were protective factors and ethnicity was a risk factor (defined by BMI, Mongolian nationality: PR 1.042, 95% CI 1.030-1.054; defined by waist circumference, PR 1.043, 95% CI 1.033-1.054) for overweight or obese participants. There were other risk factors for overweight or obese participants such as high levels of education defined by BMI (PR 1.033, 95% CI 1.010-1.058) and diet score defined by waist circumference (PR 1.004, 95% CI 1.000-1.008). After adjustment, BMI and waist circumference were associated with the greatest likelihood of systolic and diastolic hypertension (for BMI >=30 kg/m2, PR 2.455, 95% CI 1.786-3.374; for waist circumference >=88 cm, PR 1.517, 95% CI 1.133-2.031). BMI was more related to isolated diastolic hypertension than to isolated systolic hypertension, whereas waist circumference was more related to isolated systolic hypertension than to isolated diastolic hypertension. CONCLUSION: Although the prevalence of overweight and obesity as defined by BMI was low, it was relatively high as defined by waist circumference in rural Chinese women. High levels of physical activity and current smoking status had negative relations to overweight or obesity, whereas ethnicity, high levels of education, and diet score showed positive relations. Obese women defined by BMI or waist circumference had an increased risk of hypertension. PMID- 21621393 TI - Postoperative loss of skeletal muscle mass, complications and quality of life in patients undergoing cardiac surgery. AB - OBJECTIVE: The objective of this study was to describe postoperative undernutrition in terms of postoperative losses of appendicular skeletal muscle mass (ASMM) with respect to complications, quality of life, readmission, and 1-y mortality after cardiac surgery. METHODS: Patients undergoing cardiac surgery were prospectively followed. ASMM was measured 2 wk before and 2 mo after surgery using dual-energy X-ray absorptiometry. ASMM consists of arm skeletal muscle mass (SMM) and leg SMM. The association between >=5% of ASMM decrease and postoperative outcome was analyzed using the chi-square test. A similar approach was used to analyze arm SMM and leg SMM decreases separately. RESULTS: Twenty nine patients were included (23 male, 34.5% >=65 y old). Postoperatively, seven patients (24.1%) lost >=5% ASMM. When analyzed separately, a >=5% decrease in leg SMM was associated with a decrease in experienced vitality (odds ratio 13.0, 95% confidence interval 1.32-128.11, P = 0.03). In contrast, a >=5% loss of arm SMM was associated with fewer in-hospital complications (odds ratio 0.20, 95% confidence interval 0.04-0.98, P = 0.04). These patients were characterized by a higher preoperative fat-free mass index (kilograms per meter squared; P = 0.01). CONCLUSIONS: The results suggest that a preoperatively higher fat-free mass index indicates better ability to cope with operative stress, resulting in fewer complications. In addition, postoperative loss of muscle mass was associated with decreased vitality. We advocate further research investigating the effect of preoperative and postoperative nutritional intervention combined with physical exercise programs to increase lean body mass and thereby improve postoperative recovery after cardiac surgery. PMID- 21621394 TI - Efficacy of 4:1 (classic) versus 2.5:1 ketogenic ratio diets in refractory epilepsy in young children: a randomized open labeled study. AB - PURPOSE: The ketogenic (lipid to non-lipid) ratio may play an important role in the efficacy and tolerability of ketogenic diets (KD). This study was planned to compare the efficacy and tolerability of 2.5:1 versus 4:1 lipid:non-lipid ratio KD in young children with refractory epilepsy. METHODS: Children aged 6 months to 5 years with refractory epilepsy were enrolled. They were randomized to receive either a 4:1 or 2.5:1 ketogenic ratio diet, which was introduced using a non fasting protocol. Seizure frequency, biochemical profile (liver and kidney function tests, fasting lipid profile, and spot urinary calcium-creatinine ratio), and adverse effects were recorded at three months in both groups. RESULTS: Thirty eight children were enrolled, 19 in each group. At three months, 11 children (58%) in the 4:1 group and 12 (63%) in the 2.5:1 group had more than 50% reduction in seizures (p=0.78). Five children (26%) in the 4:1 group and four (21%) in 2.5:1 group became seizure free. There was no significant difference in the biochemical parameters between the two groups. CONCLUSION: 2.5:1 ratio KD is possibly as effective as 4:1 KD in controlling seizures and has fewer adverse effects. PMID- 21621395 TI - GABBR1 gene polymorphism(G1465A)isassociated with temporal lobe epilepsy. AB - PURPOSE: gamma-Aminobutyric acid B receptor 1(GABBR1) gene G1465A polymorphism has been considered as a potential risk factor for the development of temporal lobe epilepsy (TLE). However, the results were inconsistent. In this study, we performed a meta-analysis to assess the association between GABBR1 G1465A polymorphism and the risk of TLE. METHODS: Biomedical literature databases including PubMed, ISI web of science and Embase were searched. The studies evaluating the association between GABBR1 G1465A polymorphism and TLE were included. Pooled odds ratio (OR) and 95%CI confidence interval (CI) were calculated using fixed- or random-effects model. KEY FINDINGS: Seven studies (1011 cases and 2184 controls) met the inclusion criteria and were included in the meta-analysis. The overall result showed that the association between GABBR1 G1465A polymorphism was statistically significant (OR=5.381, 95%CI: 1.726, 16.776, P=0.004). Subgroup analysis showed that the effect estimate was higher in the studies with high quality score (OR=14.220, 95%CI: 6.933, 29.169, P=0.000) than that in the studies with low quality score (OR=1.158, 95%CI: 0.325, 4.123, P=0.821). SIGNIFICANCE: The present meta-analysis suggests that GABBR1 G1465A polymorphism is associated with the risk of TLE. The role of GABBR1 G1465A polymorphism in the development of TLE merits further investigation. PMID- 21621396 TI - Natural contact and stigma towards schizophrenia in African Americans: is perceived dangerousness a threat or challenge response? AB - This research extends our understanding of the relationship of social contact theory to stigma in a sample of mainly African American college students. In Phase 1, 75 participants with high levels of contact reported significantly lower negative affect and less social distance toward the mentally ill as compared to 89 participants with low contact. Despite this, the high contact group attributed significantly higher levels of dangerousness to the mentally ill. Thus while social contact was associated with reductions in some dimensions of stigma, it was associated with higher levels of self-reported perceived dangerousness. These results were obtained while controlling for social desirability bias in the self report measures of stigma. The results from Phase 1 of this study may indicate fundamental differences between incidental social contact and that which occurs in an assistive context. In Phase 2, a subset of convenience from the high contact group (n=27) and the low contact group (n=38) were compared on cardiovascular reactivity measures while imagining social interactions with people labeled with schizophrenia. Post hoc testing revealed that when participants from the high contact group imagined interacting with people labeled as schizophrenic they exhibited significant decreases in total peripheral resistance (TPR), the challenge pattern, compared to their reactions when they imagined interacting with unstigmatized people. This finding suggests the higher dangerousness ratings of the mentally ill sometimes found in African American samples may be related to factors other than direct threat. PMID- 21621397 TI - Elafin and secretory leukocyte protease inhibitor stimulate the production of cytokines and chemokines by human keratinocytes via MAPK/ERK and NF-kappaB activation. PMID- 21621398 TI - Changes in wetting properties of alumina surface treated with DPPC in the presence of phospholipase A2 enzyme. AB - Wetting properties of commercial Al(2)O(3) plates contacted with dipalmitoylphosphatidylcholine (DPPC) or DPPC+enzyme (phospholipase PLA(2)) in NaCl solution were determined by thin layer wicking and with the help of Washburn equation. Van Oss et al.'s approach to interfacial free energy interactions was applied to determining the solid surface free energy components. Wicking experiments were performed both for bare and alumina plates precontacted overnight with the probe liquid saturated vapours, as well as the untreated and DPPC (or DPPC+PLA(2)) treated alumina plates. For this purpose the penetration rates of n-octane, water and formamide were measured. From these experiments it resulted that original alumina surface is strongly polar with electron-donor interactions originating from the surface hydroxyl groups. Adsorption of DPPC on Al(2)O(3) plates slightly increased the hydrophobic character of the alumina surface (considerable decrease of the electron-donor, gamma(s)(-) parameter and gamma(s)(AB) component was visible) in such a way that the hydrocarbon chains were directed outwards and the polar part towards the alumina surface. However, after the enzyme action the products of DPPC hydrolysis by PLA(2) (palmitic acid and lysophosphatidylcholine) increased again the hydrophilic character of Al(2)O(3) surface (a minor increase in gamma(s)(AB) component and drastic increase of the electron-donor gamma(s)(-) parameter was noticeable). After treatment with DPPC or DPPC+enzyme PLA(2) solution the changes of the total surface free energy of alumina and its Lifshits-van der Waals (gamma(s)(LW)) component were in the range 7-10 mJ/m(2), but the most considerable and delivering more interesting information were the changes of the electron-donor (gamma(s)(-)) parameter ranging from 27 to 35 mJ/m(2). Moreover, the changes of the alumina surface wettability were dependent on the time of the enzyme contacting with DPPC in NaCl solution. On the basis of the obtained results it seems that the thin layer wicking method can be an additional useful tool in investigations of the effect of phospholipid and PLA(2) action on the hydrophilic hydrophobic character of alumina surface. PMID- 21621399 TI - Electrocatalytic oxidation of L-tryptophan using copper hexacyanoferrate film modified gold nanoparticle graphite-wax electrode. AB - A novel copper hexacyanoferrate (CuHCF) film modification on cysteamine (Cys) gold nanoparticle (AuNp) graphite-wax (GW) composite electrode was achieved for the quantitative determination of L-Tryptophan (L-Trp) at a reduced overpotential of 400mV in comparison with the bare Cys-AuNp-GW composite electrode. This modified electrode exhibited a well resolved pair of redox peaks corresponding to the hexacyanoferrate (II/III) reactions of CuHCF film at a formal potential of 0.65 V at a scan rate of 20 mV s(-1). Electrochemical impedance spectroscopy (EIS) studies with the modified electrode showed a very low charge transfer resistance to the electron transfer kinetics of Fe(II)/Fe(III) reactions. A linear range of 8.5*10(-7) M to 1.2*10(-4) M with a detection limit of 1.85*10( 8) M was achieved for the determination of L-Trp with a sensitivity of 0.1198 MUA/MUM. The influence of ultrasonication on the stability of the CuHCF film modified electrode was investigated. In addition, the CuHCF film modified electrode displayed an excellent reproducibility towards the real time analysis of L-Trp in commercial milk samples. PMID- 21621400 TI - Instance-based classifiers applied to medical databases: diagnosis and knowledge extraction. AB - OBJECTIVE: The aim of this paper is to study the feasibility and the performance of some classifier systems belonging to family of instance-based (IB) learning as second-opinion diagnostic tools and as tools for the knowledge extraction phase in the process of knowledge discovery in clinical databases. MATERIALS AND METHODS: We consider three clinical databases: one relating to the differential diagnosis of erythemato-squamous diseases, the second to the diagnosis of the onset of diabetes mellitus and the third dealing with a problem of diagnostic imaging in nuclear cardiology. We apply five IB classifiers to each database; two are based on exemplars, one is based on prototypes and two are hybrid. One of the latter classifiers is a new classifier introduced here and is called prototype exemplar learning classifier (PEL-C). We use cross-validation techniques to evaluate and compare the performances of several classifier systems as diagnostic tools, considering indexes such as accuracy, sensitivity, specificity, and conciseness of class representations. Moreover we analyze the number and the type of instances that represent the diagnostic classes learnt by each classifier to evaluate and compare their knowledge extraction capabilities. RESULTS: An examination of the experimental results shows that classifiers with the best classification performances are the optimized k-nearest neighbour classifier (k NNC) and PEL-C. The k-NNC uses the highest number of representative instances, 100% of the entire database, whereas PEL-C uses a far lesser number of representative instances: equal, on the average, to the 3% of the database. As tools for knowledge extraction, we interpret the kind of class representations obtained by IB classifiers as a form of nosological knowledge. Additionally, we report the most interesting diagnostic class representations to be those extracted by PEL-C because they are composed of a mixture of abstracted prototypical cases (syndromes) and selected atypical clinical cases. CONCLUSION: This study shows that IB methods - most notably, the optimized k-NNC and the PEL C - can be used and may be advantageous for clinical decision support systems and that IB classifiers can be used for nosological knowledge extraction. Because PEL C uses more compact and potentially meaningful class descriptions, it is preferable when the diagnostic problem at-hand needs smaller storage space or for knowledge extraction itself. The complexity and responsibility of diagnostic practice requires that these results be confirmed further within other clinical domains. PMID- 21621401 TI - DNA detection using commercial mobile phones. AB - This study investigates the feasibility of using mobile phones cameras for DNA detection. DNA amplification uses the convective polymerase chain reaction (cPCR) technique due to its simple mechanism, which requires no thermal cycling control. Fluorescence increment analysis and information entropy analysis are employed separately to determine whether the test samples contain target DNA (Positive) or not (Negative). The fluorescence increment method uses the brightness of the captured images before and after DNA amplification to calculate DeltaF. DeltaF values above a threshold level indicate that the test sample is positive. The information entropy method defines the probability, P(C/X), which indicates whether the fluorescence image tends towards a specific shape. If a DNA template is successfully amplified, the captured fluorescence image should be a perfect circle. P(C/X) provides a threshold of 0.5 to identify a circle and values above 0.5 indicate the test sample is positive. Experimental results show that P(C/X) is more effective than DeltaF for determining DNA detection results. The information entropy analysis method is applied to ten mobile phones of three different brands equipped with camera sensors, which have pixel numbers ranging from 120 M to 800 M. The clinical evaluation study (n = 60) for screening hepatitis B virus (HBV) plasmid samples shows that the accuracy rate of all models of mobile phones ranges from 85% to 100%. This illustrates that successful DNA detection can be achieved using the most widely deployed electronic device. PMID- 21621402 TI - Ferrocenemonocarboxylic-HRP@Pt nanoparticles labeled RCA for multiple amplification of electro-immunosensing. AB - A multiple amplification immunoassay was proposed to detect alpha-fetoprotein (AFP), which was based on ferrocenemonocarboxylic-HRP conjugated on Pt nanoparticles as labels for rolling circle amplification (RCA). Firstly, the capture antibody (anti-AFP) was immobilized on glass carbon electrode (GCE) deposited nano-sized gold particles. After a typical immuno-sandwich protocol, primary DNA was immobilized by labeling secondary antibody, which acted as a precursor to initiate RCA. The products of RCA provide large amount of sites to link detection DNAs, which were labeled by signal probes (ferrocenemonocarboxylic) and horseradish peroxidase (HRP). Moreover, the enzymatic amplification signals could be produced by the catalysis of HRP and Pt nanoparticles with the addition of H2O2. These lead to multiple amplification signals monitoring by electrochemical instrument and further resulted in high sensitivity of the immunoassay with the detection limit of 1.7 pg/mL. PMID- 21621403 TI - A novel amperometric immunosensor based on acetone-extracted propolis for the detection of the HIV-1 p24 antigen. AB - A novel amperometric immunosensor for the detection of the p24 antigen (p24Ag) from HIV-1 was constructed using gold nanoparticles (GNP), multi-walled carbon nanotubes (MWCNTs), and an acetone-extracted propolis film (AEP). First, amino functionalized MWCNTs (MWCNTNH2) were prepared and dispersed in an HAuCl4 solution to synthesize GNPs in situ. Next, the GNP/CNT/AEP nanocomposite was prepared by mixing an AEP solution and the GNP/CNT powder. The nanocomposite was dripped onto a gold electrode (GE), and then p24 antibody (anti-p24 Ab) was immobilized on the resulting modified gold electrode to construct the immunosensor. The assembly process was characterized using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The factors that were likely to influence the performance of the proposed immunosensor were studied in detail. Under optimal conditions, the proposed immunosensor exhibited good electrochemical sensitivity to the presence of p24 in a concentration range of 0.01 to 60.00 ng/mL, with a relatively low detection limit of 0.0064 ng/mL (S/N = 3). Moreover, the proposed immunosensor showed a rapid (<= 18 s) and highly sensitive amperometric response (0.018 and 1.940 MUA/ng/mL) to p24 with acceptable stability and reproducibility. PMID- 21621404 TI - Determination of trace 2,4-dinitrophenol in surface water samples based on hydrophilic molecularly imprinted polymers/nickel fiber electrode. AB - A rapid, sensitive and selective electrochemical method was proposed for the determination of 2,4-dinitrophenol (2,4-DNP) in surface water samples, using hydrophilic molecular imprinted polymers (MIPs) as the recognition element and nickel (Ni) fiber as the catalytic element. Hydrophilic MIPs were synthesized using 2,4-DNP as the template, acrylamide as the monomer, glycidilmethacrylate as the pro-hydrophilic co-monomer and acetonitrile as the solvent. Hydrophilic modification could enhance the accessibility of 2,4-DNP to the imprinted cavities and improve the selective recognition properties of traditional MIPs in water medium. Subsequently, hydrophilic MIPs/Ni fiber electrode was prepared to determine trace 2,4-DNP by cyclic voltammetry. The parameters affecting the analytical performance were investigated. Under optimized conditions, the linear range was 0.7-30 MUg L-1 and the detection limit was 0.1 MUg L-1. Finally, the proposed method was applied to measure 2,4-DNP in surface water samples. The spiked recoveries were changed from 91.3% to 102.6% and the RSD was not higher than 5.1%. There was no statistically significant difference between the results obtained by the proposed method and the traditional chromatographic method. PMID- 21621405 TI - Homogeneous detection of concanavalin A using pyrene-conjugated maltose assembled graphene based on fluorescence resonance energy transfer. AB - In this work, we proposed a novel biosensor to homogeneously detect concanavalin A (ConA) using pyrene-conjugated maltose assembled graphene based on fluorescence resonance energy transfer (FRET). Maltose-grafted-aminopyrene (Mal-Apy) was synthesized and characterized by mass spectra, UV-vis and fluorescence spectra. The Mal-Apy was further employed for fluorescence switch and ConA recognition. When Mal-Apy was self-assembled on the surface of graphene by means of pi stacking interaction, its fluorescence was adequately quenched because the graphene acted as a "nanoquencher" of the pyrene rings due to FRET. As a result, in the presence of ConA, competitive binding of ConA with glucose destroyed the pi-stacking interaction between the pyrene and graphene, thereby causing the fluorescence recovery. This method was demonstrated the selective sensing of ConA, and the linear range is 2.0 * 10-2 to 1.0 MUM with the linear equation y=1.029x + 0.284 (R = 0.996). The limit of detection for ConA was low to 0.8 nM, and the detection of ConA could be performed in 5 min, indicating that this method could be used for fast, sensitive, and selective sensing of ConA. Such data suggests that the graphene FRET platform is a great potential application for protein-carbohydrate studies, and would be widely applied in drug screening, bimolecular recognition and disease diagnosis. PMID- 21621406 TI - Loss of lamin A/C expression in stage II and III colon cancer is associated with disease recurrence. AB - AIM OF THE STUDY: Loss of the nuclear lamina protein lamin A/C (LMNA) has been observed in several human malignancies. The present study aimed to investigate associations between LMNA expression and clinical outcome in colon cancer patients. PATIENTS AND METHODS: Clinicopathological data and formalin-fixed paraffin embedded tissues were collected from 370 stage II and III colon cancer patients. Tissue microarrays were constructed, stained for lamin A/C and evaluated microscopically. Microsatellite instability status was determined for 318 tumours. RESULTS: Low levels of LMNA expression were observed in 17.8% of colon tumours, with disease recurrence occurring in 45.5% of stage II and III colon cancer patients with LMNA-low expressing tumours compared to 29.6% of patients with LMNA-high expressing tumours (p=0.01). For stage II patients, disease recurrence was observed for 35.7% of LMNA-low compared to 20.3% of LMNA high expressing tumours (p=0.03). Microsatellite stable (MSS) tumours exhibited more frequently low LMNA expression than microsatellite instable (MSI) tumours (21% versus 9.8%; p=0.05). Interestingly, disease recurrence among LMNA-low and LMNA-high expressing MSS tumours varied significantly for stage III patients who had not received adjuvant chemotherapy (100% versus 37.8%; p<0.01) while no such difference was observed for patients who received adjuvant chemotherapy (46.7% versus 46.0%; p=0.96). CONCLUSION: These data indicate that low expression of LMNA is associated with an increased disease recurrence in stage II and III colon cancer patients, and suggest that these patients in particular may benefit from adjuvant chemotherapy. PMID- 21621407 TI - The inhibition of pancreatic cancer invasion-metastasis cascade in both cellular signal and blood coagulation cascade of tissue factor by its neutralisation antibody. AB - Tissue factor (TF), the initiating cell surface receptor for the blood coagulation cascade, plays an important role in malignant transformation of the pancreas, although the precise mechanism remains unresolved. Here, we report that the TF - factor VIIa complex in human pancreatic cancer cells produced a significant amount of MMP-9 and promoted invasion ability in vitro and invasion and metastasis in vivo. For treatment, we successfully developed an anti-human TF monoclonal antibody that inhibits both cellular signalling and blood coagulation cascade via TF. Invasive capability and MMP-9 expression were significantly reduced by the antibody. The antibody inhibited not only tumour invasion in the orthotopic model, but also haematogenous metastasis in the portal-injection liver metastasis model. In conclusion, the TF-VIIa complex plays an important role in invasion-metastasis by enhancing tumour cell infiltration ability and forming microthrombi. The newly established anti-human TF neutralisation antibody may be useful for the treatment of pancreatic and other invasive cancers. PMID- 21621408 TI - The Patient Reported Outcomes Following Initial treatment and Long term Evaluation of Survivorship registry: scope, rationale and design of an infrastructure for the study of physical and psychosocial outcomes in cancer survivorship cohorts. AB - 'Patient Reported Outcomes Following Initial treatment and Long term Evaluation of Survivorship (PROFILES)' is a registry for the study of the physical and psychosocial impact of cancer and its treatment from a dynamic, growing population-based cohort of both short and long-term cancer survivors. PROFILES contains a large web-based component and are linked directly to clinical data from the population-based Eindhoven cancer registry. This paper describes the rationale and design of PROFILES. The primary aims of studies that use the PROFILES registry are: (1) psychosocial risk and outcome assessment to identify patients at high risk for poor physical and mental health outcomes, (2) to analyse mediating mechanisms to better understand the biological and behavioural factors associated with cancer treatment outcomes, and (3) to evaluate physical and psychosocial care needs of cancer survivors. PROFILES is a tool that enables data collection management; from inviting patients to participation in studies, to collecting patient-reported outcomes data via web-based or mailed questionnaires and linking these data with clinical data. The availability of a control cohort of approximately 2000 persons from the general population who complete the same basic questionnaire annually will provide the opportunity to estimate the unique impact of cancer, beyond that of normal ageing and comorbidities. Raw data from the PROFILES registry will be available for non commercial scientific research, subject to study question, privacy and confidentiality restrictions, and registration (www.profilesregistry.nl). PMID- 21621409 TI - Production of biodiesel from mixed waste vegetable oil using an aluminium hydrogen sulphate as a heterogeneous acid catalyst. AB - Al(HSO(4))(3) heterogeneous acid catalyst was prepared by the sulfonation of anhydrous AlCl(3). This catalyst was employed to catalyze transesterification reaction to synthesis methyl ester when a mixed waste vegetable oil was used as feedstock. The physical and chemical properties of aluminum hydrogen sulphate catalyst were characterized by scanning electron microscopy (SEM) measurements, energy dispersive X-ray (EDAX) analysis and titration method. The maximum conversion of triglyceride was achieved as 81 wt.% with 50 min reaction time at 220 degrees C, 16:1 molar ratio of methanol to oil and 0.5 wt.% of catalyst. The high catalytic activity and stability of this catalyst was related to its high acid site density (-OH, Bronsted acid sites), hydrophobicity that prevented the hydration of -OH group, hydrophilic functional groups (-SO(3)H) that gave improved accessibility of methanol to the triglyceride. The fuel properties of methyl ester were analyzed. The fuel properties were found to be observed within the limits of ASTM D6751. PMID- 21621410 TI - Generation of novel radiolabeled opiates through site-selective iodination. AB - Tritiated opioid radioligands have proven valuable in exploring opioid binding sites. However, tritium has many limitations. Its low specific activity and limited counting efficiency makes it difficult to examine low abundant, high affinity sites and its disposal is problematic due to the need to use organic scintillants and its relatively long half-life. To overcome these issues, we have synthesized both unlabeled and carrier-free radioiodinated iodobenzoyl derivatives of 6beta-naltrexamine ((125)I-BNtxA, 18), 6beta-naloxamine ((125)I BNalA, 19) and 6beta-oxymorphamine ((125)I-BOxyA, 20) with specific activities of 2100Ci/mmol. To optimize the utility of the radioligand, we designed a synthesis in which the radiolabel is incorporated in the last synthetic step, which required the selective iodination of the benzoyl moiety without incorporation into the phenolic A ring. Competition studies demonstrated high affinity of the unlabelled compounds for opioid receptors in transfected cell lines, as did the direct binding of the (125)I-ligands to the opioid receptors. The radioligand displayed very high sensitivity, enabling a marked reduction in tissue, as well as excellent signal/noise characteristics. These new (125)I-radioligands should prove valuable in future studies of opioid binding sites. PMID- 21621411 TI - Practical synthesis, anticonvulsant, and antimicrobial activity of N-allyl and N propargyl di(indolyl)indolin-2-ones. AB - An operation friendly protocol for the synthesis of novel di(indolyl)indolin-2 ones via Cu(OTf)(2) catalyzed bis-addition of N-allyl and N-propargyl indole with isatin was developed. This methodology allowed us to achieve the products in excellent yields without requiring purification technique like column chromatography. All the synthesized compounds were evaluated for their in vivo anticonvulsant activity against maximal electroshock test. Six compounds showed maximum activity compared to the standard drug phenytoin. The scope of the new molecules as antimicrobial agents were tested against two bacterial strains (Staphylococcus aureus and Escherichia coli) and one fungal strain (Candida albicans). PMID- 21621412 TI - Triazole-linked reduced amide isosteres: an approach for the fragment-based drug discovery of anti-Alzheimer's BACE1 inhibitors. AB - In the course of a beta-site APP-cleaving enzyme 1 (BACE1) inhibitor discovery project an in situ synthesis/screening protocol was employed to prepare 120 triazole-linked reduced amide isostere inhibitors. Among these compounds, four showed modest (single digit micromolar) BACE1 inhibition. Our ligand design was based on a potent reduced amide isostere 1, wherein the P(2) amide moiety was replaced with an anti-1,2,3-triazole unit. Unfortunately, this replacement resulted in a 1000-fold decrease in potency. Docking studies of triazole-linked reduced amide isostere A3Z10 and potent oxadiazole-linked tertiary carbinamine 2a with BACE1 suggests that the docking poses of A3Z10 and 2a in the active sites are quite similar, with one exception. In the docked structures the placement of the protonated amine that engages D228 differs considerably between 2a and A3Z10. This difference could account for the lower BACE1 inhibition potency of A3Z10 and related compounds relative to 2a. PMID- 21621413 TI - Discovery and optimization of potent and selective benzonaphthyridinone analogs as small molecule mTOR inhibitors with improved mouse microsome stability. AB - Starting from small molecule mTOR inhibitor Torin1, replacement of the piperazine ring with a phenyl ring resulted in a new series of mTOR inhibitors (as exemplified by 10) that showed superior potency and selectivity for mTOR, along with significantly improved mouse liver microsome stability and a longer in vivo half-life. PMID- 21621414 TI - Synthesis and discovery of novel hexacyclic cage compounds as inhibitors of acetylcholinesterase. AB - Hexacyclic derivatives share vital pharmacological properties, considered useful in Alzheimer's disease. The aim of this study was synthesis and its evaluation for acetyl cholinesterase inhibitory activity of novel hexacyclic analogues. Compound 4f, showed potent inhibitory activity against acetyl cholinesterase enzyme with IC(50) 0.72 MUmol/L. PMID- 21621415 TI - An alternative and expedient synthesis of radioiodinated 4-iodophenylalanine. AB - Radiolabeled amino acids have been used extensively in oncology both as diagnostic and therapeutic agents. In our pursuit to develop radiopharmaceuticals to target breast cancer, we were interested in determining the uptake of radioiodinated 4-iodophenylalanine, among other labeled amino acids, in breast cancer cells. In this work, we have developed an alternative method for the synthesis of this agent. The novel tin precursor, (S)-tert-butyl 2-(tert butoxycarbonylamino)-3-(4-(tributylstannyl)phenyl)propanoate (3) was synthesized from the known, corresponding iodo derivative. Initially, the labeled 4 iodophenylalanine was synthesized from the above tin precursor in two steps with radiochemical yields of 91.6 +/- 2.7% and 83.7 +/- 1.7% (n=5), for the radioiodination (first) and deprotection (second) step, respectively. Subsequently, it was synthesized in a single step with an average radiochemical yield of 94.8 +/- 3.4% (n=5). After incubation with MCF-7 breast cancer cells for 60 min, an uptake of up to 49.0 +/- 0.7% of the input dose was seen; in comparison, the uptake of [14C]phenylalanine under the same conditions was 55.9 +/- 0.5%. Furthermore, the uptake of both tracers was inhibited to a similar degree in a concentration-dependent manner by both unlabeled phenylalanine and 4 iodophenylalanine. With [14C]phenylalanine as the tracer, IC50 values of 1.45 and 2.50 mM were obtained for Phe and I-Phe, respectively, and these values for [125I]I-Phe inhibition were 1.3 and 1.0 mM. In conclusion, an improved and convenient method for the synthesis of no-carrier-added 4-[(*)I]phenylalanine was developed and the radiotracer prepared by this route demonstrated an amino acid transporter-mediated uptake in MCF-7 breast cancer cells in vitro that was comparable to that of [14C]phenylalanine. PMID- 21621416 TI - Resumption of JRR-4 and characteristics of neutron beam for BNCT. AB - The clinical trials of Boron Neutron Capture Therapy (BNCT) have been conducted using Japan Research Reactor No. 4 (JRR-4) at Japan Atomic Energy Agency (JAEA). On December 28th, 2007, a crack of a graphite reflector in the reactor core was found on the weld of the aluminum cladding. For this reason, specifications of graphite reflectors were renewed; dimensions of the graphite were reduced and gaps of water were increased. All existing graphite reflectors of JRR-4 were replaced by new graphite reflectors. In February 2010 the resumption of JRR-4 was carried out with new graphite reflectors. We measured the characteristics of neutron beam at the JRR-4 Neutron Beam Facility. A cylindrical water phantom of 18.6 cm diameter and 24 cm depth was set in front of the beam port with 1cm gap. TLDs and gold wires were inserted within the phantom when the phantom was irradiated. The results of the measured thermal neutron flux and the gamma dose in water were compared with that of MCNP calculation. The neutron energy spectrum of the calculation model with new reflector had little variation compared to that with old reflector, but intensities of the neutron flux and gamma dose with new reflector were rather smaller than those with old reflector. The calculated results showed the same tendency as that of the experimental results. Therefore, the clinical trials of BNCT in JRR-4 could be restarted. PMID- 21621417 TI - Place of modern imaging modalities for solitary plasmacytoma: toward improved primary staging and treatment monitoring. AB - Radiation therapy (RT) is the mainstay of treatment of solitary plasmacytoma. In most cases, doses ranging from 40 to 50 Gy yield in a local control more than 80%. However, the prognosis of patients with SP is marked by a high rate of transformation to multiple myeloma (MM), and there is no demonstrated benefit of adjuvant chemotherapy for decreasing this probability. However, clinical benefits could be reached from improving screening for other primary sites of plasmacytoma and earlier discovering signs suggestive of MM. Since such strategy could provide significant information regarding both prognosis and therapy, it has become first importance to improve initial staging of tumor widespread. Although conventional skeletal X-ray survey remains standard, usual sensitivity of radiographies does not permit diagnosing early myeloma lesions and a significant number of patients with supposed SP might be understaged and do not receive the appropriate treatment. The development of more sensitive and specific imaging modalities will make it feasible to earlier detect subclinical lesions, thus leading lead to new approaches in the treatment strategies. Here, we discuss the benefits and limitations of magnetic resonance imaging and positron emission tomography for primary staging of patients with solitary plasmacytoma. Both imaging modalities could also improve target volume delineation and assessment of tumor response after RT. PMID- 21621418 TI - Significant association of antiphospholipid antibodies and TNF-alpha: marker of severe atherogenic profile of patients with type II diabetes mellitus without micro and/or macrovascular complications. AB - OBJECTIVE: In vitro, stimulation of monocytes with antiphospholipid (aPL) antibodies resulted in increased secretion of TNF-alpha, but association of aPL with features of diabetes mellitus is not clarified yet. Therefore, we investigated the distribution of anticardiolipin (aCL), anti-beta2gpI (abeta2gpI), anti-annexin A5 (aannxA5), and anti-oxLDL (aoxLDL) antibodies, and TNF-alpha in well-formed group of 78 patients with type II diabetes mellitus without vascular complications. METHODS: Investigated antibodies and TNF-alpha concentrations were measured by ELISA. RESULTS: Antiphospholipid antibodies were in positive correlation with TNF-alpha concentrations: aCL IgG (r=0.303, p=0.007), aCL IgM (r=0.386, p=0.000), abeta2gpI IgG (r=0.499, p=0.000), abeta2gpI IgM (r=0.462, p=0.000), aanxA5 IgG (r=0.479, p=0.000), aanxA5 IgM (r=0.641, p=0.000), aoxLDL (IgG+IgM, r=0.279, p=0.000). Anticardiolipin-positive and aCL negative subgroups differed in TNF-alpha concentrations (Mann-Whitney, p=0.032). Significantly elevated LDL concentrations were noticed in aCL-positive patients with disease duration 10-15 years (chi(2)=15.000, p=0.000) and apoB concentrations were elevated in aoxLDL-positive patients with disease duration 7 10 years (chi(2)=3.938, p=0.047). CONCLUSION: Significant association of antiphospholipid antibodies and TNF-alpha might be a marker of severe atherogenic profile (suggested by increased levels of lipids in aPL-positive subgroups) and should be used for the stratification of patients with an increased risk for future deterioration of the disease. PMID- 21621419 TI - P2X7 receptor positively regulates MyD88-dependent NF-kappaB activation. AB - Recent studies have demonstrated that P2X7 plays a critical role in the immune system. Here, our results showed that P2X7 activated a NF-kappaB - but not an IFN beta-dependent luciferase reporter gene in HEK293T cells. P2X7 was involved in the LPS- and ATP-induced NF-kappaB activation but did not significantly impact the response to Zymosan in RAW264.7 cells. The activation of NF-kappaB and IFN beta induced by myeloid differentiation primary-response protein 88 (MyD88) was enhanced by P2X7 co-expression. The siRNA silencing MyD88 almost abolished the NF kappaB activation induced by P2X7, and co-immunoprecipitation showed that P2X7 interacted with MyD88. The amino acids in the C-terminus, especially the LPS binding region of P2X7, were critical for the cellular localization and immune function of P2X7. P2X7DeltaC (190 amino acids deleted in the C-terminus) and P2X7 G586A variants localized throughout the cytoplasma with a little aggregation, which differs from the cell membrane localization of wild type P2X7. Both of them could not localize to Golgi or endoplasmic reticulum. P2X7DeltaC and P2X7 G586A had impaired proteolytic cleavage of caspase-1 into the functional p20 subunit, which can activate pro-inflammatory cytokines such as IL-1beta. P2X7 G586A also showed a slight interaction with MyD88 in our co-immunoprecipitation experiment. This interaction might result in the attenuated activation of NF-kappaB and IFN beta induced by MyD88. PMID- 21621420 TI - The association between exposure to mass media and body dissatisfaction among Spanish adolescents. AB - OBJECTIVES: Correlational research has found associations between magazine and television exposure and body dissatisfaction. However, this relation is not direct, and various psychological variables may moderate and/or mediate this relation, such as awareness and internalization of the thin ideals, comparing oneself with ideal body image or self-esteem, as well as the patient's body mass index (BMI). The aim of this study was to assess the association between magazine and television exposure and body dissatisfaction among Spanish adolescents by gender. A second aim was to determine the predictors of body dissatisfaction by gender. METHODS: A cross-sectional national survey of 1,165 Spanish secondary students aged 14 to 16 years was conducted. Frequency exposure and type of TV program/magazine as well as other psychological variables, such as thin-ideal internalization and self-esteem, were associated with high levels of body dissatisfaction (BD) based on a cutoff point of 16 or above on the Eating Disorder Inventory (EDI) by gender. Multiple linear regressions were used to examine associations between body dissatisfaction, mass media topic exposure, BMI, and psychological variables. RESULTS: Mass media exposure to a specific kind of content, rather than to total exposure frequency, was more associated with body dissatisfaction in females versus males. In males, five factors accounted for 35% of the total variance of body dissatisfaction and were associated with lower TV and magazine exposure to fitness topics and lower self-esteem and with greater BMI, disordered eating behaviors, and awareness of the thin ideal. For females, high body dissatisfaction was associated with greater internalization of the thin ideal, disordered eating behaviors, BMI, and lower self-esteem (54% of total variance explained). CONCLUSION: There does not seem to be a clear association between media exposure and body dissatisfaction. Further research is necessary to enhance our understanding of how the media's role affects adolescents' awareness and internalization of the thin ideals, which may in turn lead to the development of body dissatisfaction and unhealthy weight control behaviors. PMID- 21621421 TI - The 27th annual Charles T. Dotter lecture: Data integration in interventional radiology: the pressing challenge of our time. PMID- 21621422 TI - The effectiveness of United Network of Organ Sharing status 2 transplantation in the modern era. AB - BACKGROUND: The continued benefit of United Network of Organ Sharing (UNOS) status 2 transplantation in the modern era has been questioned. METHODS: We measured deterioration to higher status designations, improvement allowing delisting, and risk of death or delisting as too ill, regardless of subsequent status, from the Scientific Registry of Transplant Recipients database. Extended Cox models were used to assess the relative hazard of status 2 transplantation vs waiting after status 2 listing. The likelihood of transplantation was measured with logistic regression. RESULTS: We analyzed 14,153 candidates listed from 2003 to 2008. Within 1 year of initial listing, deterioration to status 1B occurred frequently (63%), while delisting as too well occurred rarely (2%-7%). Death or delisting as too ill occurred among 27% at 2 years after initial status 2 listing. Mortality at 2 years after status 2 transplantation was 13%. The hazard ratio (HR) after 180 days of status 2 transplantation vs waiting during or after initial status 2 listing was 0.41 (95% confidence interval, 0.31-0.55). The likelihood of transplantation was markedly diminished for women (odds ratio, 0.71; p < 0.001) and congenital heart disease (odds ratio, 0.24; p < 0.001). Death or delisting as too ill for women (HR, 1.7; p < 0.001) and congenital heart disease (HR, 3.2; p < 0.001) were substantially higher than in other groups. CONCLUSIONS: Escalation of UNOS status is common and delisting as too well is uncommon after initial status 2 listing. Despite the decreasing number of transplants provided to status 2 registrants, sub-groups of patients may be at high risk of waiting at status 1A, justifying the continued use of the status 2 designation. PMID- 21621423 TI - Survival after biventricular assist device implantation: an analysis of the Interagency Registry for Mechanically Assisted Circulatory Support database. AB - BACKGROUND: Patients requiring biventricular assist device (BiVAD) for mechanical circulatory support (MCS) have substantially worse outcomes than patients requiring left VAD (LVAD) support only. Patient-specific risk factors have yet to be consistently identified in a large, multicenter registry, which may underlie the poorer outcomes for BiVAD patients. The Interagency Registry for Mechanically Assisted Circulatory Support (INTERMACS) is a registry of U.S. Food and Drug Administration-approved durable MCS devices used for bridge-to-transplantation, destination therapy, or recovery. The purposes of this study were to 1) identify the underlying pre-implant characteristics of the population requiring BiVAD support that contribute to reduced survival, and 2) identify differences in postoperative outcomes with respect to adverse events compared with patients supported with LVAD alone. METHODS: From June 2006 to September 2009, 1,646 patients were entered into the INTERMACS database in which adverse events and outcomes were recorded for primary implants with LVAD or BiVAD. Competing outcomes methodology was used to estimate the time-related probability of death, transplant, or recovery. Overall survival for all groups was analyzed with Kaplan Meier methods and Cox proportional regression analysis. RESULTS: The distribution of primary device implants included 1,440 LVADs and 206 BiVADs. BiVAD patients presented with a lower INTERMACS profile 93% in INTERMACS 1 or 2, compared with 73% for LVAD patients (p < 0.001). Survival at 6 months was 86% for LVADs and 56% for BiVADs (p < .0001). Adverse event rates, expressed as episodes/100 patient months for the BiVAD group compared with LVAD, were significantly higher for infection (33.2 vs 14.3), bleeding (71.6 vs 15.5), neurologic events (7.9 vs 2.6), and for device failure (4.9 vs 2.0). CONCLUSIONS: Patients requiring BiVAD support at the time of durable MCS implant are more critically ill at the time of MCS implant. BiVAD patients experience worse survival than patients supported with LVAD alone and higher rates of serious adverse events. Characteristics of the population present at the time of BiVAD implant likely influence post-implant MCS outcomes. PMID- 21621424 TI - Cardiac allograft hypertrophy is associated with impaired exercise tolerance after heart transplantation. AB - BACKGROUND: Exercise performance, an important aspect of quality of life, remains limited after heart transplantation (HTx). This study examines the effect of cardiac allograft remodeling on functional capacity after HTx. METHODS: The total cohort of 117 HTx recipients, based on echocardiographic determination of left ventricle mass and relative wall thickness at 1 year after HTx, was divided into 3 groups: (1) NG, normal geometry; (2) CR, concentric remodeling; and (3) CH, concentric hypertrophy. Cardiopulmonary exercise testing was performed 5.03 +/- 3.08 years after HTx in all patients. Patients with acute rejection or significant graft vasculopathy were excluded. RESULTS: At 1 year post-HTx, 30% of patients had CH, 55% had CR and 15% had NG. Exercise tolerance, measured by maximum achieved metabolic equivalents (4.62 +/- 1.44 vs 5.52 +/- 0.96 kcal/kg/h), normalized peak Vo(2) (52 +/- 14% vs 63 +/- 12%) and Ve/Vco(2) (41 +/ 17 vs 34 +/- 6), was impaired in the CH group compared with the NG group. A peak Vo(2) <=14 ml/kg/min was found in 6%, 22% and 48% of patients in the NG, CR and CH groups, respectively (p = 0.01). The CH pattern was associated with a 7.4-fold increase in relative risk for a peak Vo(2) <=14 ml/kg/min compared with NG patients (95% confidence interval 1.1 to 51.9, p = 0.001). After multivariate analysis, a 1-year CH pattern was independently associated with a reduced normalized peak Vo(2) (p = 0.018) and an elevated Ve/Vco(2) (p = 0.035). CONCLUSIONS: The presence of CH at 1 year after HTx is independently associated with decreased normalized peak Vo(2) and increased ventilatory response in stable heart transplant recipients. The identification of CH, a potentially reversible mechanism of impairment in exercise capacity after HTx, may have major clinical implications. PMID- 21621425 TI - Two hands are better than one: a new assessment method and a new interpretation of the non-visual illusion of self-touch. AB - A simple experimental paradigm creates the powerful illusion that one is touching one's own hand even when the two hands are separated by 15 cm. The participant uses her right hand to administer stimulation to a prosthetic hand while the Examiner provides identical stimulation to the participant's receptive left hand. Change in felt position of the receptive hand toward the prosthetic hand has previously led to the interpretation that the participant experiences self-touch at the location of the prosthetic hand, and experiences a sense of ownership of the prosthetic hand. Our results argue against this interpretation. We assessed change in felt position of the participant's receptive hand but we also assessed change in felt position of the participant's administering hand. Change in felt position of the administering hand was significantly greater than change in felt position of the receptive hand. Implications for theories of ownership are discussed. PMID- 21621426 TI - Seizure frequency and social outcome in drug resistant epilepsy patients who do not undergo epilepsy surgery. AB - Little is known about the long term prognosis of refractory epilepsy patients who do not undergo epilepsy surgery. We performed a telephone survey and chart review of patients who underwent presurgical evaluation in our Unit but did not have surgery, from 1998 until 2004. We contacted 84 patients; mean follow-up was 6.7 years. Four patients (4.7%) had died, presumably of SUDEP. Ten patients (13.1%) were seizure free. In most patients with seizures, frequency remained stable (24/80, 30%) or had decreased by >=50% (26 patients, 30.9%). Most patients (69, 86.2%) believed their health was similar or better respect to the moment they underwent presurgical evaluation. Employment situation was stable in 64/80 patients (80%), but 11 had received new disability wages. Family situation was also generally unchanged (69/80 patients, 86.2%). Most patients were not taking antidepressants. Seizure free patients scored higher in satisfaction with life. This information can be used to counsel refractory patients. PMID- 21621427 TI - A case of hypocalcemia-related epilepsia partialis continua. AB - Epilepsia partialis continua (EPC) is a syndrome clinically defined as continuous spontaneous jerking confined to one part of the body, sometimes aggravated by action or sensory stimuli, occurring over hours, days or even years. In adults the more frequent recognized cause of EPC is an acute cerebrovascular disease. Acute severe hypocalcemia is a highly epileptogenic ionic disturbance, abnormally increasing neuronal excitability. In this short communication we describe the first probable case of acute hypocalcemia-related EPC. Eight months after a left parietal lobe cardioembolic stroke, a 74-year-old woman experienced a generalized tonic-clonic seizure for the first time in her life, at the beginning of a Clostridium difficile enterocolitis. Four days later, while the abdominal symptoms were clinically improving, continuous semi-rhythmic jerks of right face, shoulder and arm began suddenly. Despite several appropriated antiepileptic treatments those involuntary movements did not cease. On routine biochemical examination we noted a total calcium serum level of 1.2 mmol/L (normal range 2.1 2.8 mmol/L), not previously known. After intravenous calcium gluconate supplementation, the jerks started to fade, disappearing completely as a total calcium serum level of 1.9 mmol/L was reached. Two separated CT brain scans did not reveal new cerebral lesions. Neurophysiological studies did not show any cortical activity related to jerks. Taken together, the treatment refractoriness and the clinical improvement after ionic imbalance correction point towards a highly possible role of hypocalcemia in sustaining the activity of a previously silent epileptogenic focus. PMID- 21621428 TI - Epileptic seizures in neurofibromatosis type 1 are related to intracranial tumors but not to neurofibromatosis bright objects. AB - OBJECTIVE: To investigate the relationship between intracranial lesions and epileptic seizures in neurofibromatosis type 1 (NF1) patients. BACKGROUND: NF1 is one of the most common autosomal dominant neurocutaneous disorders, and epilepsy is more prevalent in NF1 patients than in the general population. Epileptic seizures were found to be related to various types of intracranial lesions in NF1 patients. METHODS: The clinical characteristics of NF1 (1986-2006 in Chung-Gung Memorial Hospital), diagnosed on the basis of the criteria of the National Institutes of Health Consensus Conference (1988), were reviewed by 2 neurologists. We diagnosed epileptic seizures of NF1 patients on the basis of clinical appearances and a history of antiepileptic drugs. Magnetic resonance images were also evaluated by 2 neuroradiologists to confirm the locations of brain tumors or neurofibromatosis bright objects (NBOs). The locations of NBOs were classified into 4 categories: cortex and hippocampus, subcortical white matter, basal ganglia, and infratentorial area. The association between the location of the lesions and the occurrence of seizure in NF1 patients was analyzed statistically. RESULTS: The medical records of 630 NF1 patients were reviewed. In this cohort, 37 (5.87%) NF1 patients had epileptic seizures. The patients include 22 males (59.5%) and 15 females (40.5%). The mean seizure onset age was 14.8 years (2 months to 72 years). The most common seizure pattern was partial onset seizures, 3 simple partial seizures, and 14 complex partial seizures. Other seizure types found include 15 primary generalized seizures (2 absence seizures and 13 generalized tonic-clonic seizures), 2 infantile spasms, and 3 unclassified. A total of 172 (23 with epilepsy and 149 without epilepsy) NF1 patients underwent MRI examinations. NBOs were identified in 16 (69.6%) epilepsy patients and in 108 (72.5%) patients without epilepsy. The location or the number of these intracranial lesions does not show significant correlation with the occurrence of epilepsy in our cohort. Among 11 NF1 patients with intracranial tumors, 4 patients had seizures (36.36%), vs. 19 out of 161 NF1 patients (11.80%) without tumors. CONCLUSION: The occurrence of epileptic seizures in NF1 patients is related to intracranial tumors but not to NBOs. PMID- 21621429 TI - Nodular excision for painful localized Achilles tendon xanthomas in type II hyperlipoproteinemia: a case report. AB - Achilles tendon xanthomas are often associated with type II hyperlipoproteinemia, in which low-density lipoprotein derived from the circulation accumulates in the tendons. Sometimes coronary artery disease can jeopardize the life of the patient if the condition is neglected. We describe the case of bilateral painful Achilles tendon xanthomas in a heterozygous type II hyperlipoproteinemia family. Her symptoms were not alleviated despite anti-inflammatory medication and eccentric exercise for 6 months. She was treated with nodular excision of the xanthomas bilaterally and then with postoperative statins to avoid recurrence. PMID- 21621430 TI - Combined total ankle replacement and modified bridle tendon transfer for end stage ankle joint arthrosis with paralytic dropfoot: report of an unusual case. AB - In recent years, total ankle replacement has become a reasonable option for many patients with end-stage ankle arthrosis. In order to be successful, total ankle replacement requires a relatively balanced alignment of the foot in relation to the leg. Such alignment is traditionally achieved surgically by means of stabilization of the hindfoot in conjunction with relocation osteotomy of the calcaneus and/or tibia. In this report, we describe the unconventional combination of total ankle replacement in an adult patient with concomitant paralysis that was addressed by means of tendon transfer. PMID- 21621431 TI - Interphalangeal dislocation of toes: a retrospective case series and review of the literature. AB - Although not uncommon, dislocation of the toes, including that of the great toe, is not commonly reported in published studies. In the present report, we describe a series of 18 patients with toe dislocations managed by our department from January 2001 to December 2007. We considered the radiographic pattern of injury in our series of patients. Of the 18 patients, 10 (55.56%) had their toe dislocations treated by closed reduction with or without internal fixation. Seven patients (38.89%) with complex dislocation, defined as open dislocation or dislocation not amenable to (failed attempt) closed reduction, that required open reduction and internal fixation. One patient (5.56%) with a dislocated toe declined to undergo any form of treatment. PMID- 21621432 TI - Post-treatment leukocytosis predicts an unfavorable clinical response in patients with moderate to severe diabetic foot infections. AB - Our aim was to determine whether post-treatment laboratory values could help to predict the clinical response in patients with advanced diabetic foot infections. One hundred and three consecutive patients hospitalized for moderate or severe diabetic foot infections at a large, university-affiliated hospital were identified and their records retrospectively reviewed. Definitive therapy during each patient's hospital course was defined as any foot surgery when additional major surgery was not anticipated or when a course of deep soft tissue and/or bone culture-specific antibiotics had been initiated. The clinical response was assessed at 90 days after the start of definitive therapy. A poor response was recognized as persistent infection at the initial or a contiguous site or when unplanned revision surgery or amputation was subsequently required. The peripheral white blood cell count, neutrophil count, erythrocyte sedimentation rate, and C-reactive protein levels measured shortly after initiating definitive therapy (i.e., post-treatment) were examined for their association with the clinical response using logistic regression models. A total of 38 patients with the complete compliment of laboratory and clinical follow-up aged 59.7 +/- 12.3 years with a diabetes duration of 13.3 +/- 9.1 years were included. Leukocytosis, defined as a white blood cell count >11,000 cells/MUL, observed an average of 3 +/- 1.4 days after treatment, was the single most important marker for predicting a poor clinical response, and the only significant study variable in both univariate and multivariate analyses (multivariate odds ratio 9.7, 95% confidence interval 1.0 to 92, p = .048). We conclude that leukocytosis observed shortly after initiating definitive therapy is predictive of an unfavorable clinical response by 90 days. PMID- 21621433 TI - Medial fracture line significance in calcaneus fracture. AB - In Sanders' classification of calcaneus fractures, the medial fracture line (subtype C) is close to the tarsal canal, which contains an artery for the talus and calcaneus. We hypothesized that because of this brittle vascular localization, patients with C line fracture patterns might describe radiologic subtalar arthritis more often and have more complaints. The purpose of the present study was to compare the results of C line fracture patterns with other types of calcaneus fractures. A total of 25 surgically treated feet were involved. Regarding Sanders' classification, group 1 included fractures involving the C line (11 feet), and group 2 included fractures not involving the C line (14 feet). Patient age at admission, trauma date, and interval until surgery were obtained from the patients' medical records. The Bohler angles were determined from the radiographs. At the last follow-up visit, the radiologist graded subtalar arthritis using computed tomography. For clinical follow-up, the American Orthopaedic Foot and Ankle Society and Maryland scores were assessed. No significant differences were found in mean age, follow-up period, delay to surgery, or postoperative Bohler angle between the 2 groups. The mean preoperative Bohler angle was significantly low for group 1. Although not significantly different, the mean American Orthopaedic Foot and Ankle Society and Maryland scores were lower for group 1 (81.9 and 84.3) than group 2 (87.8 and 92.0), and the median subtalar arthritis grade was greater for group 1 (score 2) than for group 2 (score 1.5). The worse results with C line fracture patterns despite satisfactory reduction might result from sinus tarsi artery damage. Angiographic investigations could clarify this theory in the future. Consequently, surgeons must inform and should hesitate to operate on patients with these highly comminuted C line calcaneus fractures. PMID- 21621434 TI - Axial loading cross screw fixation for the Austin bunionectomy. AB - The Austin procedure has become a common method of osteotomy for the correction of hallux abductovalgus when indicated. The V-type configuration is intrinsically stable but not without complications. One complication encountered is rotation and/or displacement of the capital fragment. We present the use of an axial loading screw in conjunction with a dorsally placed compression screw. The benefit to this technique lies in the orientation of the axial loading screw, because it is directed to resist the ground reactive forces while also providing a second point of fixation in a crossing screw design. In a head-to-head biomechanical comparison, we tested single dorsal screw fixation versus double screw fixation, including both the dorsal and the axial loading screws in 10 metatarsal Sawbones((r)) (Pacific Research Laboratories Inc, Vashon, WA). Five metatarsals received single dorsal screw fixation and five received the dorsal screw and the additional axial loading screw. The metatarsals were analyzed on an Instron compression device for comparison; 100% of the single screw fixation osteotomies failed with compression at an average peak load of 205 N. Four of five axial loading double screw fixation osteotomies did not fail. This finding suggests that the addition of an axial loading screw providing cross screw orientation significantly increases the stability of the Austin osteotomy, ultimately decreasing the likelihood of displacement encountered in the surgical repair of hallux abductovalgus. PMID- 21621435 TI - Lateral subtalar dislocation associated with bimalleolar fracture: case report and literature review. AB - Subtalar dislocation is an uncommon injury that affects the talocalcaneal and talonavicular joints, with the tibiotalar and calcaneocuboid joints remaining intact. The 4 types of subtalar dislocation are medial, lateral, anterior, and posterior, although the latter 2 are rare. These injuries-especially lateral dislocation-occur as a result of high-energy trauma. Medial dislocation is the most common type, and lateral dislocation is associated with osteochondral fractures of the talus and calcaneus, as well as with open injuries; hence, its worse prognosis. We report the case of a 62-year-old woman diagnosed with lateral subtalar dislocation accompanied by an ipsilateral bimalleolar fracture after a fall downstairs. She underwent emergency reduction of the dislocation under sedation. Surgical treatment of the bimalleolar fracture was delayed 9 days to avoid cutaneous complications. This is the first report of a subtalar dislocation accompanied by a bimalleolar fracture. PMID- 21621436 TI - Uncommon fusion of teeth and lateral periodontal cyst in a Chinese girl: a case report. AB - Fusion is a developmental anomaly of dental hard tissues. Most cases occur in the anterior region of both the primary and permanent dentitions. Occasionally, fusion of permanent and supernumerary teeth may occur in the maxillary anterior region; however, a supernumerary tooth fused with a premolar is a rare case. Developmental lateral periodontal cyst is also an uncommon developmental odontogenic cyst with no clinical symptoms, and the lesion is often discovered on routine radiographic examination. In the present case, we report the uncommon fusion of teeth and lateral periodontal cyst in a Chinese girl. Using cone beam computed tomography we are able to acquire a better understanding of the complicated root canal morphology of the fused tooth and successfully manage the lateral periodontal cyst. PMID- 21621437 TI - Keeping an eye on ourselves. PMID- 21621438 TI - The contribution of the mitochondrial COI/tRNA(Ser(UCN)) gene mutations to non syndromic and aminoglycoside-induced hearing loss in Polish patients. AB - Mutations in mitochondrial DNA have been implicated in both, non-syndromic and aminoglycoside-induced hearing loss. In the present study, we have performed the systematic mutation screening of the COI/tRNA(Ser(UCN)) genes in 250 unrelated Polish subjects with hearing impairment. Three different homoplasmic sequence variants were identified, including one common polymorphism m.7476 C>T in tRNA(Ser(UCN)) and two mutations, m.7444 G>A and m.7445 A>G localized in the COI/precursor of tRNA(Ser(UCN)). The incidence of m.7444 G>A substitution was estimated at 1.6% (4/250), however variable penetrance of hearing loss, age of onset and hearing thresholds among m.7444 G>A carriers was observed. Two subjects had the positive history of aminoglycoside exposure and one of them harbored both m.7444 G>A and 12S rRNA m.1555 A>G mutations. Those suggest that m.7444 G>A itself is not sufficient to produce a clinical phenotype and additional modifier factors are required for pathogenic manifestation of m.7444 G>A substitution. Moreover, we have described the first Polish family with non-syndromic hearing loss, harboring m.7445 A>G mutation. The penetrance of hearing loss in this pedigree was 58% when aminoglycoside-induced hearing impairment was included, and 8% when ototoxic effect was excluded. This finding strongly suggests the possible role of m.7445 A>G in susceptibility to aminoglycoside induced-hearing loss. PMID- 21621439 TI - Glucocerebrosidase mutations do not cause increased Lewy body pathology in Parkinson's disease. AB - Mutations in the glucocerebrosidase (GBA) gene have been implicated in increased formation of Lewy bodies (LBs) in Parkinson's disease (PD). We found GBA mutation status not to be significantly associated with the density of cortical LBs, after adjusting for sex, age at death, duration of PD and presence of dementia. Comparison of GBA carriers to PD controls found no difference in Alzheimer's disease pathological findings. Our results do not support GBA carriers to have a more advanced neuropathologic disease i.e. increased density of protein aggregates. PMID- 21621440 TI - [Quality assessment for preanalytical phase in clinical laboratory: a multicentric study]. AB - PURPOSE: To show the number of preanalytical sample errors in seven laboratories attending seven health departments of Valencian Community (Spain). METHODS: Cross sectional study of the number of preanlytical errors in samples obtained in primary care centers. An error is defined as a rejected specimen: any blood or urine sample, which cannot be successfully tested as it does not meet the acceptability criteria of the laboratory or if the sample is not received. We collected preanalytical errors from the tests requested for hematology, coagulation, chemistry, and urine samples. Registers were collected and indicators calculated automatically through a data warehouse and OLAP cubes software. RESULTS: Large differences in the results of preanalytical errors were observed between health departments. The highest percentage of errors occurred in coagulation samples, followed by urine, hematology and biochemistry. With regard to the type of error, the largest proportion of errors was due to failures of process. CONCLUSIONS: The high incidence of preanalytical errors and variability between health departments suggests that there is a need to standardize the drawing practice. PMID- 21621441 TI - Acute neonatal suppurative parotitis: a case report and review of the literature. AB - Neonatal suppurative parotitis (NSP) is an uncommon disease. Information about the etiopathogenesis and management of the disease is very limited. Here, we describe a newborn who developed NSP due to Pseudomonas aeruginosa and who was treated successfully with antibiotics. PMID- 21621442 TI - Detection of Leishmania parasites in the blood of patients with isolated cutaneous leishmaniasis. AB - BACKGROUND: The consequences of the spread of Leishmania parasites to the blood from lesions in patients with cutaneous leishmaniasis are numerous. To assess the magnitude of this invasion we conducted the present study on patients referred to the American University of Beirut Medical Center for cutaneous leishmaniasis. METHODS: Patients referred for the management of cutaneous leishmaniasis were included in the study. Skin and blood cultures for Leishmania were taken from these patients. RESULTS: One hundred sixty-two patients were proven to have cutaneous leishmaniasis by pathology; 52% were males and 44% females (gender information was missing for 4%). Patient age ranged from 5 months to 70 years. None of the patients had received treatment for Leishmania. We obtained parasite isolates from 85 patients (52.5%), proven by cultures from skin and blood/blood components. Interestingly, the parasite was isolated in the blood and blood components of 50 patients (30.9%). Isoenzyme analysis confirmed the fact that the organisms in blood and skin were the same; from the 28 isolates that were positive in both skin and blood, eight isolates were Leishmania major and two were Leishmania tropica. The remaining isolates, whether positive in both blood and skin or in either of these tissues, skin or blood and its products, were Leishmania infantum sensu lato. CONCLUSIONS: In the current study, the detection rate of parasites in the blood of patients with cutaneous leishmaniasis was high. This illustrates the invasive characteristic of the parasite that has escaped the skin. Testing should be considered in areas other than Lebanon, especially around the Mediterranean basin. Whether these findings support the administration of systemic treatment for cutaneous leishmaniasis or not needs to be confirmed in larger prospective studies. PMID- 21621443 TI - [About a fortuitous representation of a case of intestinal malrotation by Wilhem His (1880)]. AB - In a textbook published in 1882, His exhibits drawings devoted to intestine development. The last figure depicts a typical case of gut malrotation, but there is no comment about this in the text. It could be possible that this omission should be due to the fact that mechanisms of gut development were not totally elucidated at this time. PMID- 21621444 TI - [An anatomical wax of the deep structures of the pelvic limb (by Tramond, 19th century): observation of the tridimensional photographic rotation]. AB - Anatomical wax modelling was widely used during the 19(th) century, especially in France and Italy. In Paris, The Tramond house was specialized in the realization of this kind of samples. MATERIAL AND METHODS: The sample was placed on two large horizontal marble disks, rotating every 5 degrees . We could then describe the sample, verify its anatomical accuracy and also perform a virtual reconstruction with Quick Time Reality QTVR((r)) software. RESULTS: The muscular, nervous and arterial elements were represented. We divided the description in three parts: (1) lumbar, pelvic and femoral; (2) femoro-tibial; and (3) tibio-tarsian. We focused the anatomical description on the sciatic nerve; on the organization of the muscles of the gluteal region and the neurovascular organization; and on arterial segmentation. This sample was getting damaged with time, noticeably the representation of the nerves, which are very thin and so, very fragile. Nowadays, 3D representation of the dissected human body is more common, with new techniques such as plastination (Von Hagen's type), which allows one to preserve all the anatomical elements of the subject. This paper and the realization of the virtual museum also aim to participate in a work memory, which recognize the knowledge of the anatomist of wax makers, their teaching quality remained unequalled as their obvious artistical value. PMID- 21621445 TI - Diagnostic criteria for generalized osteoarthritis: a preliminary study in a population with knee osteoarthritis. PMID- 21621446 TI - [An optimum donor site for venous grafting for microsurgery]. AB - We report the results of an anatomic study based on 10 cadavers. The aim of this work is to find an optimum donor site for venous grafts which is safe, reproducible, and suitable for microsurgery stitches especially in finger reimplantation, when a long and small calibre graft is needed. This study describes the deep venous network of the radial artery, an original donor site for microsurgical venous grafts. The second aim is to describe our technique of harvesting. Dissections always show two satellite veins, that can be harvested with optimal average diameter of 1.8mm constant over the whole length. The maximum length available is about 126.5mm for the radial satellite vein, and 125 mm for the ulnar one, with a few number of collateral ligatures needed. No tying is required in 60% cases for radial satellite vein, and one ligature for the other 40%, whereas in the ulnar satellite vein, no tying is needed in 80% and just one in the other 20%. This original site is advantageous in microsurgery of the upper limb, offering an easy, quick, safe and reproducible option in an emergency situation. PMID- 21621447 TI - Gunshot injuries to the nerves. AB - The authors report their 10-year experience in the treatment of war injuries in the Gaza strip. Bullet injuries to nerves can be assessed using Sunderland's classification. Grade 4 and 5 lesions must be repaired in the same way as any traumatic nerve injuries. A detailed series of sciatic nerve repair is presented. In this series, 12 nerve gaps were repaired by direct nerve coaptation using 90 degrees knee flexion for six weeks. The results of this technique at 1 to 4 years follow-up are better than to those of sciatic nerve grafting. PMID- 21621448 TI - Is particulate air pollution associated with health and health inequalities in New Zealand? AB - Air pollution can increase mortality risk and may also exacerbate socioeconomic inequalities in health outcomes. This New Zealand study investigated whether exposure to particulate air pollution (PM(10)) was associated with mortality and health inequalities. Annual mean PM(10) estimates for urban Census Area Units (CAUs) were linked to cause-specific mortality data. A dose-response relationship was found between PM(10) and respiratory disease mortality, including at concentrations below the existing annual average guideline value of 20MUgm(-3). Establishing and enforcing a lower guideline value is likely to have population health benefits. However, socioeconomic inequalities in respiratory disease mortality were not significantly elevated with PM(10) exposure. PMID- 21621449 TI - On the convergence of EM-like algorithms for image segmentation using Markov random fields. AB - Inference of Markov random field images segmentation models is usually performed using iterative methods which adapt the well-known expectation-maximization (EM) algorithm for independent mixture models. However, some of these adaptations are ad hoc and may turn out numerically unstable. In this paper, we review three EM like variants for Markov random field segmentation and compare their convergence properties both at the theoretical and practical levels. We specifically advocate a numerical scheme involving asynchronous voxel updating, for which general convergence results can be established. Our experiments on brain tissue classification in magnetic resonance images provide evidence that this algorithm may achieve significantly faster convergence than its competitors while yielding at least as good segmentation results. PMID- 21621450 TI - (18)FDG-PET/CT for detecting distant metastases and second primary cancers in patients with head and neck cancer. A meta-analysis. AB - We performed a meta-analysis to evaluate the role of (18)F-fluorodeoxyglucose positron emission tomography/computed tomography ((18)FDG-PET/CT) in detecting distant metastases and second primary cancers in patients with head and neck cancer (HNC) at staging. Studies about (18)FDG-PET/CT for detecting distant metastases and second primary cancers in patients with HNC were systematically searched in the MEDLINE, EMBASE, and EBM Review databases from January 1, 2000 to March 1, 2011. A software called "Meta-DiSc" was used to obtain pooled estimates of sensitivity, specificity, diagnostic odds ratios, summary receiver operating characteristic curves, and Q* index. Twelve articles fulfilled all inclusion criteria. The pooled sensitivity, specificity, and Q* index estimates with 95% confidence interval for PET/CT were 0.888 (0.827-0.928), 0.951 (0.936-0.963), and 0.937 (0.844-0.964). Subgroup analysis showed similar pooled estimates of sensitivity, specificity, and Q* index for initial staging and restaging, nasopharyngeal cancer and all other sites of HNC. (18)FDG-PET/CT has good diagnostic performance in detecting distant metastases and second primary cancers in patients with HNC. PMID- 21621451 TI - An ecological perspective on U.S. industrial poultry production: the role of anthropogenic ecosystems on the emergence of drug-resistant bacteria from agricultural environments. AB - The industrialization of food animal production, specifically the widespread use of antimicrobials, not only increased pressure on microbial populations, but also changed the ecosystems in which antimicrobials and bacteria interact. In this review, we argue that industrial food animal production (IFAP) is appropriately defined as an anthropogenic ecosystem. This paper uses an ecosystem perspective to frame an examination of these changes in the context of U.S. broiler chicken production. This perspective emphasizes multiple modes by which IFAP has altered microbiomes and also suggests a means of generating hypotheses for understanding and predicting the ecological impacts of IFAP in terms of the resistome and the flow of resistance within and between microbiomes. PMID- 21621452 TI - Spectroscopic and quantum chemical studies on 4-acryloyl morpholine. AB - Fourier transform infrared (FTIR) and FT-Raman spectra have been recorded and an extensive spectroscopic investigations have been carried out on 4-acryloyl morpholine (4AM). Theoretical quantum chemical studies have also been performed. From the ab initio and DFT analysis using HF, B3LYP and B3PW91 methods with 6 31G(d,p) and 6-311G++(d,p) basis sets the energies, structural, thermodynamical and vibrational characteristics of the compound were determined. The energy difference between the chair equatorial and chair axial conformers of 4AM have been calculated by density functional theory (DFT) method. The optimized geometrical parameters, theoretical wavenumbers and thermodynamic properties of the molecule are compared with the experimental values. The effect of acryloyl group on the characteristic frequencies of the morpholine ring has been analysed. The mixing of the fundamental modes with the help of potential energy distribution (PED) through normal co-ordinate analysis has been discussed. PMID- 21621453 TI - Computerized detection of adverse drug reactions in the medical intensive care unit. AB - OBJECTIVE: Clinical event monitors are a type of active medication monitoring system that can use signals to alert clinicians to possible adverse drug reactions. The primary goal was to evaluate the positive predictive values of select signals used to automate the detection of ADRs in the medical intensive care unit. METHOD: This is a prospective, case series of adult patients in the medical intensive care unit during a six-week period who had one of five signals presents: an elevated blood urea nitrogen, vancomycin, or quinidine concentration, or a low sodium or glucose concentration. Alerts were assessed using 3 objective published adverse drug reaction determination instruments. An event was considered an adverse drug reaction when 2 out of 3 instruments had agreement of possible, probable or definite. Positive predictive values were calculated as the proportion of alerts that occurred, divided by the number of times that alerts occurred and adverse drug reactions were confirmed. RESULTS: 145 patients were eligible for evaluation. For the 48 patients (50% male) having an alert, the mean+/-SD age was 62+/-19 years. A total of 253 alerts were generated. Positive predictive values were 1.0, 0.55, 0.38 and 0.33 for vancomycin, glucose, sodium, and blood urea nitrogen, respectively. A quinidine alert was not generated during the evaluation. CONCLUSIONS: Computerized clinical event monitoring systems should be considered when developing methods to detect adverse drug reactions as part of intensive care unit patient safety surveillance systems, since they can automate the detection of these events using signals that have good performance characteristics by processing commonly available laboratory and medication information. PMID- 21621454 TI - Hepatitis C virus core antigen: analytical performances, correlation with viremia and potential applications of a quantitative, automated immunoassay. AB - BACKGROUND: Testing for hepatitis C virus core antigen (HCV Ag) may represent a complementary tool to anti-HCV and HCV-RNA in the diagnosis and monitoring of HCV infection. OBJECTIVE: To evaluate the performance characteristics of the automated Abbott ARCHITECT HCV Ag assay. STUDY DESIGN: Five sites analyzed over 3000 routine serum samples from populations at different risk, comparing HCV Ag results with anti-HCV screening and supplemental assay results and with HCV-RNA. RESULTS: The HCV Ag assay showed a specificity of 100%, a good precision (CV<10%) and excellent dilution linearity (r>0.999). The sensitivity (3 fmol/L) corresponds to 700-1100 IU/mL of HCV-RNA. A non-linear correlation with HCV-RNA was found: r=0.713 vs. Siemens bDNA (523 specimens), r=0.736 vs. Roche Cobas TaqMan (356 specimens) and r=0.870 vs. Abbott Real-Time PCR (273 specimens). HCV Ag quantitation was equally effective on different HCV genoypes (239 for genotype 1/1a/1b/1c, 108 for genotype 2/2a/2c, 86 for genotype 3/3a, 50 for genotype 4/4a/4c/4d). Testing of subjects at high risk for HCV and with potential or actual impairment of the immune system identified 2 cases negative for anti-HCV and positive for HCV Ag on 361 hemodialyzed (0.6%) and 7 cases on 97 (7.2%) among transplant recipients. HCV Ag positivity anticipated anti-HCV seroconversion in all three cases of acute hepatitis C. CONCLUSIONS: HCV Ag may be used as reflex testing on anti-HCV positive individuals to confirm or exclude an active infection, and on subjects with acute hepatitis or belonging to high risk groups. PMID- 21621455 TI - Ultrastructure and LSU rDNA-based phylogeny of Peridinium lomnickii and description of Chimonodinium gen. nov. (Dinophyceae). AB - Several populations of Peridinium lomnickii were examined by SEM and serial section TEM. Comparison with typical Peridinium, Peridiniopsis, Palatinus and Scrippsiella species revealed significant structural differences, congruent with phylogenetic hypotheses derived from partial LSU rDNA sequences. Chimonodinium gen. nov. is described as a new genus of peridinioids, characterized by the Kofoidian plate formula Po, cp, x, 4', 3a, 7'', 6c, 5s, 5''', 2'''', the absence of pyrenoids, the presence of a microtubular basket with four or five overlapping rows of microtubules associated with a small peduncle, a pusular system with well defined pusular tubes connected to the flagellar canals, and the production of non-calcareous cysts. Serial section examination of Scrippsiella trochoidea, here taken to represent typical Scrippsiella characters, revealed no peduncle and no associated microtubular strands. The molecular phylogeny placed C. lomnickii comb. nov. as a sister group to a clade composed of Thoracosphaera and the pfiesteriaceans. Whereas the lack of information on fine structure of the swimming stage of Thoracosphaera leaves its affinities unexplained, C. lomnickii shares with the pfiesteriaceans the presence of a microtubular basket and the unusual connection between two plates on the left side of the sulcus, involving extra-cytoplasmic fibres. PMID- 21621456 TI - Similar relative mutation rates in the three genetic compartments of Mesostigma and Chlamydomonas. AB - Levels of nucleotide substitution at silent sites in organelle versus nuclear DNAs have been used to estimate relative mutation rates among these compartments and explain lineage-specific features of genome evolution. Synonymous substitution divergence values in animals suggest that the rate of mutation in the mitochondrial DNA is 10-50 times higher than that of the nuclear DNA, whereas overall data for most seed plants support relative mutation rates in mitochondrial, plastid, and nuclear DNAs of 1:3:10. Little is known about relative mutation rates in green algae, as substitution rate data is limited to only the mitochondrial and nuclear genomes of the chlorophyte Chlamydomonas. Here, we measure silent-site substitution rates in the plastid DNA of Chlamydomonas and the three genetic compartments of the streptophyte green alga Mesostigma. In contrast to the situation in animals and land plants, our results support similar relative mutation rates among the three genetic compartments of both Chlamydomonas and Mesostigma. These data are discussed in relation to published intra-species genetic diversity data for the three genetic compartments of Chlamydomonas and are ultimately used to address contemporary hypotheses on the organelle genome evolution. To guide future work, we describe evolutionary divergence data of all publically available Mesostigma viride strains and identify, for the first time, three distinct lineages of Mesostigma. PMID- 21621457 TI - Nanomaterials: Confusing statements by Becker et al. (2010). PMID- 21621458 TI - Exercise prescription for patients with type 2 diabetes and pre-diabetes: a position statement from Exercise and Sport Science Australia. AB - Type 2 diabetes mellitus (T2DM) and pre-diabetic conditions such as impaired fasting glucose (IFG) and/or impaired glucose tolerance (IGT) are rapidly increasing in prevalence. There is compelling evidence that T2DM is more likely to develop in individuals who are insufficiently active. Exercise training, often in combination with other lifestyle strategies, has beneficial effects on preventing the onset of T2DM and improving glycaemic control in those with pre diabetes. In addition, exercise training improves cardiovascular risk profile, body composition and cardiorespiratory fitness, all strongly related to better health outcomes. Based on the evidence, it is recommended that patients with T2DM or pre-diabetes accumulate a minimum of 210 min per week of moderate-intensity exercise or 125 min per week of vigorous intensity exercise with no more than two consecutive days without training. Vigorous intensity exercise is more time efficient and may also result in greater benefits in appropriate individuals with consideration of complications and contraindications. It is further recommended that two or more resistance training sessions per week (2-4 sets of 8-10 repetitions) should be included in the total 210 or 125 min of moderate or vigorous exercise, respectively. It is also recommended that, due to the high prevalence and incidence of comorbid conditions in patients with T2DM, exercise training programs should be written and delivered by individuals with appropriate qualifications and experience to recognise and accommodate comorbidities and complications. PMID- 21621459 TI - Pulmonary emphysematous changes in patients with congenital heart disease associated with increased pulmonary blood flow: evaluation using multidetector row computed tomography. AB - BACKGROUND: The present study aimed to evaluate the prevalence and the location of segmental emphysematous change in congenital heart disease (CHD) patients with increased pulmonary blood flow using multidetector-row computed tomography (MDCT). METHODS: A total of 129 consecutive patients (mean age, 5.8+/-5.4 years; range, 1 month to 24 years) underwent MDCT angiography of the thorax. The frequency of emphysematous change was evaluated in patients with ventricular septal defect (VSD, n=61), atrial septal defect (ASD, n=27), patent ductus arteriosus (PDA, n=36) and complete atriventriclar septal defect (CAVSD, n=5). In 59 patients who underwent cardiac catheterisation, the relationships between the emphysematous change and both pulmonary to systemic blood flow ratio (Qp/Qs) and mean pulmonary arterial pressure (mPAP) were evaluated. RESULTS: The emphysematous change was detected in 57 patients (44.2%) out of 129 patients. The frequency of segmental emphysematous change in left side was higher than in right side (14.8% vs. 6.5%). Both Qp/Qs and mPAP affected the presence of emphysema. CONCLUSION: MDCT can provide accurate detection of segmental emphysema in patients with CHD. Emphysematous change is not uncommon pathological lesion in children and adolescents with CHD. PMID- 21621460 TI - "I'm still who I was" creating meaning through engagement in art: The experiences of two breast cancer survivors. AB - PURPOSE: The aim of this feasibility study was two-fold: i) develop lifelike torsos of two breast cancer survivors using innovative sculpting material and; ii) shed light on the meaning women give to the experience of breast cancer after viewing their sculpted torsos. METHODS: This collaborative initiative between nurse researchers and artists was situated within phenomenological inquiry. Two breast cancer survivors shared their bodies, as models, and stories of their cancer journey and the experience of modeling to create life-sized torsos of their bodies. KEY RESULTS: The participants articulated embodied knowing as each shared experiences of connectedness and relationship which culminated in the emergence of four core themes: The Landscape of Breast in Cancer; Red Shoes: The Re-claiming of Self; Liberation: towards an embodied self; and, Scars: Re authoring Life. CONCLUSIONS: Active engagement in art through the use of one's body has the potential to open the door to healing, the generation of meaning and a reaffirmation of self. PMID- 21621461 TI - Evolution of the arthropod nervous system. Part 2. PMID- 21621462 TI - Efficacy and safety of palliative chemotherapy for patients with advanced breast cancer pretreated with anthracyclines and taxanes: a systematic review. AB - No standard monotherapy or combination palliative chemotherapy currently exists for patients with advanced breast cancer pretreated with anthracyclines and taxanes. In this systematic review we assess the current knowledge on the efficacy and safety of palliative single-agent chemotherapy drugs--capecitabine, vinorelbine, gemcitabine, and liposomal doxorubicin--commonly used in daily clinical practice. We identified 22 studies, of which ten investigated capecitabine, nine investigated vinorelbine, three investigated gemcitabine, and one investigated liposomal doxorubicin. The greatest amount of information was available for capecitabine and vinorelbine. These two drugs showed good efficacy. The disease control rate differed significantly between the four drugs, which is relevant in terms of how well tumour symptoms can be improved and whether quality of life can be maintained or even improved. To obtain more evidence of the efficacy and safety of chemotherapeutic agents used in this pretreated population of advanced breast cancer patients, randomised comparisons of the various drugs, as monotherapy and in combination with targeted agents, are needed. PMID- 21621463 TI - The experience of general nurses in rural Australian emergency departments. AB - Australia is a geographically unique country with large areas classed as rural. Nurses providing emergency care in rural hospitals face a number of challenges, with rural communities expecting multi-skilled nurses, prepared for a wide range of unannounced situations. Using a mixed method approach, involving questionnaires and focus groups, the study was undertaken in two rural health services in Victoria, Australia. The aim was to explore the experiences of general nurses working in rural hospital settings, with regards to their emergency department responsibilities. The findings indicate that nurses lacked confidence, which they attributed to the sporadic nature of working in the area and the diversity of people who presented. A resultant 'skills rusting' was described and nurses identified the need to be a diverse 'specialist'. Some lack of confidence, particularly in the mental health area, was related to feelings of isolation and lack of context specific education and training. While some excellent emergency specific education and training is available for rural nurses, access is limited by a multitude of constraints. This study found there is an urgent need for local emergency education and training; with nurses showing a strong preference for ongoing professional development incorporating scenario based and context specific education. PMID- 21621464 TI - Increased frequency and cell death of CD16+ monocytes with Mycobacterium tuberculosis infection. AB - Monocytes from tuberculosis patients exhibit functional and phenotypical alterations compared with healthy controls. To determine whether these discrepancies can be explained by changes in monocyte subsets, the expression of CD14 and CD16 was evaluated in tuberculosis patients and healthy controls; additionally, some markers related to the mononuclear phagocytes maturation, differentiation and function, such as CD1a, CD1c, CD11b, CD11c, CD13, CD33, CD36, CD40, CD64, CD68, CD80, CD83, CD86, HLA-DR, CCR2, CCR5, and non-specific esterases (NSE) were determined in monocyte subsets. Patients had increased percentage of circulating CD14(Hi)CD16(+) and CD14(Lo)CD16(+) monocytes. The percentage of monocytes expressing CD11b, CD36, CD64, CD68, CD80, CD86, CCR2 and NSE was lower in CD14(Hi)CD16(+) and CD14(Lo)CD16(+) cells than in CD14(Hi)CD16( ) monocytes. M. tuberculosis infected CD16(+) monocytes produced more TNF-alpha and less IL-10 than CD16(-) cells at 6 h post-infection. Isolated CD16(+) monocytes spontaneously underwent apoptosis during differentiation into macrophages; in contrast to CD16(-) monocytes that became differentiated into monocyte-derived macrophages (MDM) with a minimal induction of cell death. In addition, there were more Annexin V and propidium iodide positive monocytes in the CD16(+) subset infected with live M. tuberculosis at 24 h than CD16(-) monocytes. Under the culture conditions established for this study, the monocyte subsets did not differentiate into dendritic cells. These results show that tuberculosis patients have an augmented frequency of CD16(+) circulating monocytes which are more prone to produce TNF-alpha and to undergo cell death in response to M. tuberculosis infection. PMID- 21621465 TI - Assessing the effect of pneumococcal conjugate vaccines: what is the value of routinely collected surveillance data? PMID- 21621466 TI - Herd immunity and serotype replacement 4 years after seven-valent pneumococcal conjugate vaccination in England and Wales: an observational cohort study. AB - BACKGROUND: The seven-valent pneumococcal conjugate vaccine (PCV7) has reduced vaccine-type (VT) invasive pneumococcal disease but increases in non-vaccine-type (NVT) disease have varied between countries. We assess the effect of the PCV7 vaccination on VT and NVT disease in England and Wales. METHODS: The study cohort was the population of England and Wales from July, 2000, to June, 2010. We calculated incidence rate ratios (IRRs) to compare incidences of VT and NVT disease before (2000-06) and after (2009-10) the introduction of PCV7. We used data from the national surveillance database. Cases included in our analysis were restricted to those confirmed by culture linked with isolates referred for serotyping at the national reference centre by laboratories in England and Wales. We adjusted for potential bias from missing data (serotype and age of patient) and changes in case ascertainment rates during the study period. FINDINGS: 5809 cases of invasive pneumococcal disease were reported in 2009-10, giving an incidence of 10.6 per 100,000 population in 2009-10, which, when compared with the adjusted average annual incidence of 16.1 in 2000-06, gives an overall reduction of 34% (95% CI 28-39). VT disease decreased in all age groups, with reductions of 98% in individuals younger than 2 years and 81% in those aged 65 years or older. NVT disease increased by 68% in individuals younger than 2 years and 48% in those aged 65 years or older, giving an overall reduction in invasive pneumococcal disease of 56% in those younger than 2 years and 19% in those aged 65 years or older. After vaccine introduction, more NVT serotypes increased in frequency than decreased, which is consistent with vaccine-induced replacement. Key serotypes showing replacement were 7F, 19A, and 22F. Increases in NVT invasive pneumococcal disease were not associated with antimicrobial resistance. INTERPRETATION: Despite much serotype replacement, a substantial reduction in invasive pneumococcal disease in young children can be achieved with PCV7 vaccination, with some indirect benefit in older age groups. Further reductions should be achievable by use of higher valency vaccines. Robust surveillance data are needed to properly assess the epidemiological effect of multivalent pneumococcal disease vaccines. FUNDING: Health Protection Agency. PMID- 21621467 TI - The impact of student diversity on interest, design, and promotion of Web-based tailored nutrition and physical activity programs for community colleges. AB - OBJECTIVE: To examine an organizational level perspective of the process of adopting Web-based tailored nutrition and physical activity programs for community college students. METHODS: In this qualitative study, 21 individual key informant interviews of community college student services and health center administrators were used to examine organizational-level perceptions of interest in, design characteristics of, and ways to promote health programs. A cross classification matrix of a priori and emergent themes related to student diversity was created to describe cross-cutting patterns. RESULTS: Findings revealed 5 emergent themes for consideration in program development related to student diversity: (1) multiple roles played by students, (2) limited access to financial resources, (3) varied student demographics, (4) different levels of understanding, and (5) commuting to campus. CONCLUSIONS AND IMPLICATIONS: Nutrition and physical activity programs for community colleges need to specifically address the diverse nature of their students to increase the potential of adoption. PMID- 21621468 TI - Effects of oligodeoxynucleotide with CCT repeats on chronic graft versus host disease induced experimental lupus nephritis in mice. AB - A synthesized single-stranded oligodeoxynucleotide (ODN), designed as SAT05f with the sequence of human microsatellite DNA, has been studied for its capacity of alleviating the lupus nephritis in the chronic graft versus host disease (cGVHD) induced lupus-prone mice. In cGVHD model mice, both of continuous and discontinuous treatment with SAT05f was effective on reducing anti-ssDNA antibody production, decreasing renal IgG deposition and delaying the onset of lupus nephritis. In addition, SAT05f could down-regulate TLR9 mRNA expression in splenocytes of cGVHD model mice. These results indicated that SAT05f could be developed as a new therapeutic agent for the treatment of lupus nephritis by inhibiting TLR9 signaling pathways. PMID- 21621469 TI - Identifying research priorities in long term care homes. AB - Concerns about the quality of care in long term care (LTC) homes range from inadequate daily care to understaffing and insufficient funding. LTC decision makers are challenged to keep up with the changing demographics of residents admitted to LTC who have increasingly complex care needs. Decisions regarding LTC policies and procedures need to be informed by research that identifies the most effective and efficient care practices. This study solicited feedback from LTC decision makers in Ontario, Canada, regarding research priorities to guide improvement in the quality of care in LTC homes. Representatives from 134 LTC homes responded (53.6% response rate). Nine thematic areas of research were identified: delivery of care; staffing; organization and structure of homes; funding; indicators, standards, policies, and procedures; managing difficult behaviors; education; safety; and infectious disease control. It is anticipated that these themes will steer research down a path that is responsive to the information needs of practitioners in LTC homes. PMID- 21621470 TI - High rate of anticoagulation therapy in oldest old subjects with atrial fibrillation: the Octabaix study. AB - OBJECTIVE: To assess the prevalence of permanent atrial fibrillation (AF) in community-dwelling subjects aged 85 and to determine the anticoagulation therapy rate. DESIGN: Community-based survey of inhabitants aged 85 years. PARTICIPANTS: Participants were 328 people born in 1924 and assigned to 7 primary health care teams. MEASUREMENTS: Sociodemographic variables, Barthel Index, the Spanish version of the Mini-Mental State Examination, Mini Nutritional Assessment, Charlson Index, social risk, quality of life, and prevalent chronic diseases were assessed. Permanent AF was determined by patient interview, treatment prescriptions, and clinical records. RESULTS: The sample included 202 women (61.6%) and 126 men. In 41 (12.5%) participants permanent AF was diagnosed; 25 (60.9%) were on anticoagulant oral therapy and 9 (21.9%) were receiving antiplatelet therapy. Using multiple logistic regression analysis, previous diagnosis of heart failure (P < .001, OR 4.170, 95% CI 1.927-9.024) and stroke history (P < .03, OR 2.439, 95% CI 1.101-5.401) were significantly associated with the diagnosis of permanent AF. CONCLUSION: AF is quite prevalent in 85-year old subjects. A large percentage of patients with AF were receiving chronic anticoagulant therapy. The percentage of patients who were not receiving prophylactic treatment was low. PMID- 21621471 TI - The role of cognitive impairment in the use of the Diskus inhaler. AB - BACKGROUND AND PURPOSE: Drugs delivered by metered-dose inhalers and dry powder inhalers (DPIs) are a mainstay in the treatment of chronic lung disease; however, previous studies suggest cognitive impairment hinders proper use of inhalers. The purpose of this study was to determine the relationship between the score on the Mini-Mental State Exam (MMSE) and the ability of nursing facility residents to complete the steps required for proper use of a multiunit-dose DPI (Diskus). METHODS: Nursing facility residents who had never used a multiunit-dose DPI (Diskus), who scored between 10 and 24 inclusive on the MMSE, and who were able to hold a breath for 10 seconds were recruited for an observational study to test their ability to use a placebo-loaded Diskus when supervised and assisted by personnel trained in the proper use the Diskus. Ability to use the DPI was assessed by the Diskus Evaluation Rating Scale (DERS), an instrument developed specifically for this study. Possible scores on the DERS ranged from 0 to 19, with a score of 0 indicating no limitations in any of the steps involved in using the Diskus and 19 indicating inability to do any of the steps after 3 supervised attempts. RESULTS: Forty Diskus-naive nursing facility residents (86 +/- 9 years of age; 32 women) with MMSE scores between 10 and 24 inclusive and the ability to hold a breath for 10 seconds were enrolled in the study. Mean MMSE scores were 17.4 +/- 4.2, whereas the mean score on the DERS was 5.1 +/- 3.2 (range 1-16). After controlling for age, gender, and education, a significant inverse relationship was noted between scores on the MMSE and the DERS such that for every 1-point increase on the MMSE, the subject's DERS score decreased by 0.345 points (P = .003). Overall, 38 of the 40 subjects with MMSE scores between 10 and 24 inclusive were able to use the Diskus successfully. CONCLUSION: For MMSE scores, the better the performance on the MMSE, the better the performance on the DERS. More important, 95% of the subjects in this study could use the Diskus successfully when properly supervised. In contrast to earlier studies, these findings suggest that a multiunit-dose DPI can be prescribed as one component of the regimen for chronic lung disease in patients with substantial cognitive impairment. PMID- 21621472 TI - Geriatric conditions and adverse drug reactions in elderly hospitalized patients. AB - OBJECTIVES: To investigate the relationship between clinical conditions typically observed in the geriatric patients (geriatric conditions) and adverse drug reactions in older patients admitted to acute care hospitals. DESIGN AND SETTING: Prospective observational study conducted in 11 acute care medical wards throughout Italy. PARTICIPANTS: Five hundred six patients aged 65 years or older consecutively admitted to participating wards. MEASUREMENTS: The outcome of the study was the occurrence of any adverse drug reactions during the hospital stay. Geriatric conditions considered in the analysis were basic activities of daily living, history of falls, slow walking speed, malnutrition, dementia, depression, 1 or more unplanned admissions in the previous 3 months, history of stroke, unintentional weight loss, and exhaustion. The relationship between risk factors and outcomes was assessed using logistic regression. RESULTS: Female gender (odds ratio [OR] 2.29; 95% confidence interval [CI] 1.18-4.45) and number of medications taken during hospitalization (OR 1.12; 95% CI 1.06-1.18), but not individual Geriatric conditions, were associated with the outcome after correction for potential confounders. However, the simultaneous presence of history of falls and dependency in at least 1 activities of daily living (OR 2.18; 95% CI 1.13-4.19) was associated with adverse drug reactions during stay. CONCLUSION: The simultaneous presence of history of falls and dependency in at least one activity of daily living defines a condition of particular vulnerability of elderly hospitalized patients to adverse drug reactions. Physicians should be aware of this high-risk condition when prescribing new drugs to disabled older people. PMID- 21621473 TI - Osteoporosis and fragility fractures. PMID- 21621474 TI - Vision in the global evaluation of older individuals hospitalized following a fall. AB - INTRODUCTION: The objective of this study was to verify if vision is appropriately evaluated in older individuals admitted to a Geriatric Assessment Unit following a fall. METHODS: A retrospective clinical chart review of 158 patients from 3 university-based Geriatric Assessment Units is presented. The clinical charts of patients admitted following a fall in the Geriatric Assessment Units of 3 Montreal hospitals, between April 2006 and 2008, were reviewed. Clinical charts from age- and sex-matched controls hospitalized in the Geriatric Assessment Units during the same period but without a history of fall or fracture, were also reviewed. Pertinent sociodemographic, medical, and visual characteristics were extracted from the charts and entered into a database for analysis. RESULTS: The mean age +/- standard deviation for the cases (n = 79) and controls (n = 79) were 82.3 +/- 6.2 years and 81.7 +/- 6.4 years, respectively. Most falls were not a result of accidents, but rather were more often related to underlying medical problems that were multifactorial in origin. More cases than controls were taking antiarrhythmic and antidepressant medications, whereas more controls were taking calcium channel blockers. Cases were more likely to have cataracts, age-related macular degeneration, and decreased visual acuity. Although cases were referred more often than controls for an eye examination, they were not referred in a systematic fashion. DISCUSSION: Our results indicate that more visual problems are identified in persons who fall and, even if they are referred more often than controls for an eye examination, their vision is not evaluated systematically by an eye care specialist despite current clinical recommendations. CONCLUSION: These data indicate that eye care professionals should work more closely with the medical team to improve the overall clinical care of older individuals with a history of falls. PMID- 21621475 TI - Cholinesterase inhibitor and memantine in terminal care. PMID- 21621476 TI - Swab culture of purulent skin infection to detect infection or colonization with antibiotic-resistant bacteria. AB - Prescribing systemic antibiotics without susceptibility testing has significant shortcomings, especially in long term care facilities with high rates of multidrug-resistant organisms (MDROs) including methicillin-resistant Staphylococcus aureus. Tissue biopsy or aspiration sampling of infected tissue is the "gold standard" for culture of skin and soft tissue infection and is especially important with serious infection, systemic toxicity, or failure of initial therapy. Swab cultures are probably the most commonly used method to determine the resistance pattern of skin pathogens treated in nursing home residents. However, they are controversial, especially when obtained from chronic wounds. The culture may be obtained from an uninfected wound and lead to unnecessary antibiotic therapy. If material superficial to the infected living tissue is sampled, colonizers may be isolated. This report is focused on swab culture obtained by the Levine technique, after debridement or cleaning down to viable tissue when an acute purulent skin infection has been diagnosed based on clinical criteria. Swab cultures should not be used to determine IF a wound is acutely infected; rather the role may be to identify potential pathogens when deep tissue biopsy is not elected. The swab culture may identify the pathogen or overlying MDRO colonization, a risk factor for MDRO infection. MDRO isolation should heighten the clinician's level of concern if the prescribed antibiotic did not "cover" the MDRO or potential pathogen that was isolated. Properly performed swab cultures could play a role in the identification of methicillin-resistant Staphylococcus/MDRO infections treated in nursing homes. PMID- 21621477 TI - Prevention of venous thromboembolism in long term care: results of a multicenter educational intervention using clinical practice guidelines: part 2 of 2 (an AMDA Foundation project). AB - INTRODUCTION: Implementation of prophylaxis for venous thomboembolism (VTE) through risk assessment based on clinical practice guidelines (CPGs) is variably adopted in long term care facilities (LTCF). Current guidelines recommend venous thromboembolism prophylaxis (VTE-P) following risk assessment, individualized to patient status. In LTCF, differing comorbidity, life-expectancy, ethical, and quality-of-life issues may warrant a unique approach. This article examines VTE-P practices in LTCF before and after educational intervention to bring practice patterns consistent with CPGs. METHODS: Phase 1 (preceding article in this issue) identified current practice to assess risk and implement VTE-P (17 geographically diverse LTCFs, 3260 total beds). Phase 2 (educational intervention using CPGs) and Phase 3 (outcomes) reexamined VTE-P at the same 17 centers. RESULTS: The frequency of indications for VTE-P and contraindications to anticoagulation were similar during Phases 1 and 3 (all P > .05). In Phase 3, use of aspirin alone decreased more than 50% (P < .0005), whereas use of compression devices increased (P < .0005). Regression models predicted no relationship between any indication or contraindication and VTE-P in Phase 1 (all P > .05) but identified significant relationships between indication and contraindications and VTE-P in Phase 3 (P = .022 to P < .0005), suggesting adequate understanding of current CPGs following education as the basis for improved VTE-P. CONCLUSIONS: The study confirms the presence of significant comorbidity in LTC residents, many with indications for VTE-P, some with contraindications for anticoagulation. Following educational intervention, more residents received VTE-P, influenced by risk-benefit ratio favoring treatment. These findings suggest that even a modest educational intervention significantly improves provider knowledge pertinent to risk assessment consistent with CPG and more appropriate VTE-P. PMID- 21621478 TI - Family caregiving in advanced chronic organ failure. AB - OBJECTIVES: To assess caregiver burden as well as positive aspects of family caregiving in advanced chronic obstructive pulmonary disease (COPD), chronic heart failure (CHF), and chronic renal failure (CRF). DESIGN: Cross-sectional observational study. SETTING: Patients recruited at the outpatient clinics of academic and general hospitals in the Netherlands. PARTICIPANTS: Patients with advanced COPD (n = 73), CHF (n = 45), and CRF (n = 41) and their family caregivers. MEASUREMENTS: Caregiver burden and positive aspects of caregiving were assessed using the Family Appraisal of Caregiving Questionnaire for Palliative Care and were compared among family caregivers of patients with COPD, CHF, or CRF using linear regression analysis while controlling for characteristics of patients and family caregivers. RESULTS: Most family caregivers were female partners of participating patients. Caregiver distress and caregiver strain scores were relatively low, whereas scores for positive caregiving appraisals and family well-being were relatively positive. Caregiver strain, positive caregiving appraisals, and family well-being were comparable for family caregivers of patients with COPD, CHF, or CRF. Caregiver distress was higher for family caregivers of patients with COPD than CHF. The experience of caregiving was influenced by being the patient's spouse, patient's psychological symptoms, and the presence of comorbidities. CONCLUSIONS: Family caregiving for patients with COPD, CHF, or CRF should not only be seen as a burden, but also as a positive experience. To support family caregivers, attention should be paid to caregiver burden and the positive aspects of family caregiving. PMID- 21621479 TI - Associations between published quality ratings of skilled nursing facilities and outcomes of medicare beneficiaries with heart failure. AB - INTRODUCTION: Nursing Home Compare quality ratings are designed to allow patients, families, and clinicians to compare facilities based on quality, but associations of the current measures with important clinical outcomes are not known. Our study examined associations between ratings and readmission and mortality among Medicare beneficiaries admitted to a skilled nursing facility with a primary diagnosis of heart failure. METHODS: We conducted a retrospective cohort study of 164,672 Medicare beneficiaries discharged to skilled nursing facilities after hospitalization for heart failure in 2006-2007. The main outcome measures were readmission and mortality within 90 days. RESULTS: One-fifth of the 13,619 skilled nursing facilities received a 1-star rating and 11% received a 5 star rating. Nearly half of the patients discharged to a skilled nursing facility were readmitted to a hospital within 90 days after discharge, and 30% died within 90 days. Compared with patients in 5-star skilled nursing facilities, patients in 1-star facilities had higher risks of 90-day readmission (hazard ratio, 1.08) and mortality (1.15). After adjustment for facility size and ownership type, the associations between the quality rating and readmission were not statistically significant, but the associations with mortality were significant. CONCLUSION: Publicly reported Nursing Home Compare quality ratings of Medicare-certified skilled nursing facilities were modestly associated with 90-day readmission and mortality among Medicare beneficiaries discharged to these facilities after hospitalization for heart failure. PMID- 21621480 TI - Prevention of venous thromboembolism: practice patterns in 17 geographically diverse long term care facilities in the United States: part 1 of 2 (an AMDA Foundation project). AB - INTRODUCTION: Current guidelines recommend antithrombotic prophylaxis for venous thromboembolism (VTE) using risk assessment, factoring contraindications. This report represents a summary of current practice patterns to prevent VTE in long term care as Phase 1 of a 3-phase educational intervention study. PHASE 1 PARTICIPANTS: Participants were 376 new admissions/readmissions (77 +/- 12 [SD] years; 67% female) from 17 geographically diverse long term care facilities (3260 total beds). MEASUREMENTS: The process describes current VTE prophylaxis (VTE-P) practices; a companion article describes the educational intervention (Phase 2) and outcome (Phase 3). Phase 1 data were collected on use of nonpharmacological measures and antithrombotic drugs for VTE-P between July and September 2009. RESULTS: Indications for VTE-P were evident in 85% of new admissions, of which two-thirds received VTE-P. Contraindications for anticoagulation were observed in 54.8% of admissions, including quality of life or patient/caregiver wishes. Logistic regression analysis predicted no relationship between any indication for or any contraindication to VTE-P and use of VTE-P, suggesting an inadequate understanding of current clinical practice guidelines. CONCLUSIONS: Residents of long term care have significant comorbidity that poses risk for VTE; although many received VTE-P, contraindications were common, warranting individualized considerations. The likelihood of VTE-P was greatest following orthopedic surgery, severe trauma, and medical illness. PMID- 21621481 TI - Predictors of hospitalization in Italian nursing home residents: the U.L.I.S.S.E. project. AB - OBJECTIVE: To examine resident and facility characteristics associated with hospitalization in a cohort of Italian older nursing home residents. DESIGN: A longitudinal observational study. SETTING: The nursing homes participating in the U.L.I.S.S.E. study, a project evaluating the quality of care for older persons in Italy. SETTING PARTICIPANTS: Nursing home residents in 31 Italian nursing homes. MEASUREMENT: Each resident underwent a comprehensive geriatric assessment at baseline, and after 6 months and 1 year by means of the RAI MDS 2.0. Facility characteristics were collected using an ad hoc designed questionnaire. Hospitalizations were self-reported by facilities. RESULTS: A total of 170 (11.6%) of 1466 nursing home residents were admitted to the hospital at least once during the study period. Female gender and higher physician, nurse, and nursing assistant hours per resident were predictive of a lower probability to be admitted to the hospital, whereas a diagnosis of arrhythmia, a previous urinary tract infection, and polypharmacy were associated with a higher probability of being hospitalized. CONCLUSION: These findings suggest that a reduction of hospitalization of nursing home residents could be achieved by providing an adequate amount of care and optimizing the management of chronic diseases and polypharmacy. This hypothesis should be tested in future clinical trials. PMID- 21621482 TI - Incentivizing nursing home quality and physician performance. PMID- 21621483 TI - Metroplasty in a large population of women with septate uterus. AB - STUDY OBJECTIVE: To determine the reproductive outcome after hysteroscopic metroplasty in women with septate uterus. DESIGN: Retrospective comparative single-center study (Canadian Task Force classification II-3). SETTING: University-affiliated hospital. PATIENTS: Two hundred forty-six patients with septate uterus undergoing hysteroscopic metroplasty between January 1998 and December 2007. The diagnosis was based on hysteroscopy and 3-dimensional ultrasonography. In the most cases laparoscopy was also available. The subjects were divided into 2 age-matched groups. Group 1 consisted of 108 women with unexplained infertility, and group 2 consisted of 138 women suffering from recurrent abortion. INTERVENTIONS: Patients underwent hysteroscopic metroplasty by use of resectoscopy with an equatorial semicircular loop cutting at 0 degree with monopolar energy. All septa were completely removed without complications of bleeding, infection, risk of perforation, visceral injury, or uterine dehiscence during pregnancy. MEASUREMENT AND MAIN RESULTS: The 2 groups were compared in terms of reproductive performance in mean +/- SD follow-up of 37 +/- 18 months. In group 1, 61 (56.5%) patients achieved pregnancy. Seventy-one pregnancies ensued, including 1 twin gestation. Fourteen of the 71 pregnancies (19.7%) ended in miscarriage. In group 2, 90 (65.3%) patients achieved pregnancy. One hundred twenty-nine pregnancies ensued, including 2 twin gestations. Forty-four of the 129 pregnancies (34.1%) ended in miscarriage. The 2 groups have no significant differences in terms of reproductive outcome after surgery, except for the number of abortions, which was higher in group 2 (p <.05). CONCLUSION: This study confirms that hysteroscopic metroplasty is a simple, safe, and rapid surgical procedure with no complications for achieving normal uterine architecture, which is peculiar to a good reproductive outcome. The use of an equatorial semicircular loop may give satisfactory and similar results to those obtained with Collin's loop. PMID- 21621484 TI - Numerical simulation of molecular uptake via electroporation. AB - A numerical study of electroporation-mediated molecular delivery is presented. The model consists of the Nernst-Planck equations for species transport, coupled with an asymptotic Smoluchowski equation for membrane permeabilization. The transfer of calcium ions into a Chinese Hamster Ovary cell is simulated. The results reveal important physical insights. First, for this particular case, ion electrophoresis plays an important role, and is an order of magnitude faster than free diffusion on a comparable time scale. Second, the maximum achievable concentration within the cell is reciprocally correlated with the extracellular electrical conductivity. This behavior is mediated by an electrokinetic mechanism known as field-amplified sample stacking. Through this mechanism, the intracellular ion concentration can reach a level higher than the extracellular one provided that the intra-to-extracellular conductivity ratio is greater than unity. The results corroborate well with data in the literature, and offer a mechanistic interpretation to previous experimental observations. This work is a step toward the quantification of molecular delivery via electroporation. PMID- 21621485 TI - Application of the response surface methodology for optimization of whey protein partitioning in PEG/phosphate aqueous two-phase system. AB - In order to develop a new strategy for beta-lactoglobulin (beta-lg) removal from whey protein, partitioning of alpha-lactalbumin (alpha-la), beta-lg and glycomacropeptide (Gmp) was studied using aqueous two phase systems (ATPS). A system composed of 13% (w/w) polyethylene glycol (PEG, average molar mass 2000 g/mol) and 13% (w/w) potassium phosphate was used at 25 degrees C. A central composite rotatable design (CCRD) associated to the response surface methodology (RSM) was applied to investigate the effects of NaCl concentration and pH on the partition of these proteins. It was found that alpha-la and Gmp partitioned to the top phase rich in PEG, whereas beta-lg partitioned to the bottom phase rich in salt. According to the RSM, optimal conditions for beta-lg removal where found where pH was equal to 6.7 and salt concentration was 0.35 mol/L. Under these conditions, the partition coefficient K(alpha) was 0.48 and K(Gmp) was 0.92. On the other hand, the partition coefficient K(beta) was only 0.01. In such conditions beta-lg preferentially concentrates in the bottom phase, while the top phase exclusively contains the proteins alpha-la and Gmp. Fractionation of the proteins from fresh whey was performed in a three stage cross-flow extraction system. The extraction yield for beta-lg in the bottom phase was 97.3%, while the yields for alpha-la and Gmp in the top phase were 81.1% and 97.8%, respectively. PMID- 21621486 TI - Characterisation of a glycosylated alkyl polyglycoside produced by a cyclodextrin glycosyltransferase by HPLC-ELSD and -MS. AB - A transglycosylation reaction between an alkyl polyglycoside and alpha cyclodextrin catalysed by cyclodextrin glycosyltransferase (CGTase) from Bacillus macerans was investigated. The reaction products were identified by comparison with standards generated by CGTase catalysed modification of pure alkyl glycosides using HPLC-ELSD and -MS analysis. The main products were alkyl glucopyranosides (substrates present in the alkyl polyglycoside) glycosylated with 6 (primary coupling products) or 12 (secondary coupling products) glucose residues. Both alpha and beta anomers were glycosylated. PMID- 21621487 TI - Simultaneous analysis of synthetic musks and triclosan in human breast milk by gas chromatography tandem mass spectrometry. AB - A comprehensive method was developed for the simultaneous analysis in human breast milk of 12 synthetic musks, five nitro musks, six polycyclic muks and one macrocyclic musk; as well as one musk metabolite and triclosan. The target analytes were freeze dried and extracted using the accelerated solvent extraction (ASE) procedure. The extracts were further purified by gel permeation chromatography (GPC) and florisil solid-phase extraction (SPE) and then analyzed by gas chromatography tandem mass spectrometry (GC-MS/MS). Recoveries of the analytes based on the isotopic internal standard correction ranged from 82.4% to 112%, with relative standard derivations less than 20%. The method quantification limits (MQLs) were 0.6-5.4 ng/g lipid. The analytes were detected in human breast milk samples and ranged from 11.7 to 308.6 ng/g lipid. PMID- 21621488 TI - Extraction and preparative purification of tanshinones from Salvia miltiorrhiza Bunge by high-speed counter-current chromatography. AB - A method for extraction and preparative separation of tanshinones from Salvia miltiorrhiza Bunge was successfully established in this paper. Tanshinones from Salvia miltiorrhiza Bunge were extracted using ethyl acetate as the extractant under reflux. The extracts were then purified by high speed counter-current chromatography (HSCCC) with light petroleum-ethyl acetate-methanol-water (6:4:6.5:3.5, v/v) as the two phase solvent system. The upper phase was used as the stationary phase and the lower phase as the mobile phase. 8.2mg of dihydrotanshinone I, 5.8 mg of 1,2,15,16-tetrahydrotanshiquinone, 26.3mg of cryptotanshinone, 16.2mg of tanshinone I, 25.6 mg of neo-przewaquinone A, 68.8 mg of tanshinone IIA and 9.3mg of miltirone were obtained from 400mg of extracts from Salvia miltiorrhiza Bunge in one-step HSCCC separation, with the purity of 97. 6%, 95.1%, 99.0%, 99.1%, 93.2%, 99.3% and 98.7%, respectively, as determined by HPLC area normalization method. Their chemical structures were identified by 1H NMR. PMID- 21621489 TI - Quantitation of desmosine and isodesmosine in urine, plasma, and sputum by LC MS/MS as biomarkers for elastin degradation. AB - The aim of this study is to develop a standardized LC-MS/MS method for accurate measurement of desmosine (DES) and isodesmosine (IDS) in all body fluids as biomarkers for in vivo degradation of matrix tissue elastin in man and animals. A reproducible three-step analytical procedure: (1) sample hydrolysis in 6N HCl, (2) SPE by a CF1 cartridge with addition of acetylated pyridinoline as internal standard (IS), and (3) LC/MSMS analysis by SRM monitoring of transition ions; DES or IDS (m/z 526-481+397) and IS (m/z 471-128) was developed. The method achieves accurate measurements of DES/IDS in accessible body fluids (i.e. urine, plasma, and sputum). LOQ of DES/IDS in body fluids is 0.1 ng/ml. The % recoveries and reproducibility from urine, plasma, and sputum samples are above 99 +/- 8% (n = 3), 94 +/- 9% (n = 3) and 87 +/- 11% (n = 3), with imprecision 8%, 9% and 10%, respectively. The proposed method was applied to measure DES/IDS in body fluids of patients with chronic obstructive pulmonary disease (COPD) and healthy controls. Total DES/IDS in sputum and plasma is increased over normal controls along with the free DES/IDS in urine in patients. DES/IDS can be used to study the course of COPD and the response to therapy. This practical and reliable LC MS/MS method is proposed as a standardized method to measure DES and IDS in body fluids. This method can have wide application for investigating diseases which involve elastic tissue degradation. PMID- 21621490 TI - MultiSimplex optimization of chromatographic separation and dansyl derivatization conditions in the ultra performance liquid chromatography-tandem mass spectrometry analysis of risperidone, 9-hydroxyrisperidone, monoamine and amino acid neurotransmitters in human urine. AB - A pre-column dansylated ultra-performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-MS/MS) method for simultaneous determination of risperidone (RIP), 9-hydroxyrisperidone (9-OH-RIP), monoamine and amino acid neurotransmitters in human urine was developed with the aim of providing data on how neurotransmitters may influence each other or change simultaneously in response to risperidone treatment. MultiSimplex based on the simplex algorithm and the fuzzy set theory was applied to the optimization of chromatographic separation and dansyl derivatization conditions during method development. This method exhibited excellent linearity for all the analytes with regression coefficients higher than 0.997. The lower limit of quantification (LLOQ) values for 9-OH-RIP and RIP were 0.11 and 0.06 ng/ml, respectively, and for neurotrasmitters ranged from 0.31 to 12.8 nM. The mean accuracy ranged from 94.7% to 108.5%. The mean recovery varied between 81.6% and 97.5%. All the RSD of precision and stability were below 9.7%. Finally, the optimized method was applied to analyze the first morning urine samples of schizophrenic patients treated with risperidone and healthy volunteers. PMID- 21621491 TI - Questioning the distinction between intra- and extra-musical meaning: Comment on "Towards a neural basis for processing musical semantics" by Stefan Koelsch. PMID- 21621492 TI - Lycium barbarum: a new hepatotoxic "natural" agent? PMID- 21621493 TI - Management of gastrointestinal stromal tumours of limited size: proposals from a French panel of physicians. AB - A number of guidelines on the management of gastro-intestinal stromal tumours (GISTs) have been published, mostly based on expert consensus. However, these guidelines have generally failed to address the specific problem of GISTs of limited size (i.e. those measuring a few centimetres in diameter) with which gastroenterologists are increasingly confronted. The aim of the present work was to draw up proposals for the diagnosis and treatment of GISTs measuring less than 5 cm in diameter. For this purpose, a number of practical questions were put to a panel of French experts. PMID- 21621494 TI - Virtual enteroscopic images of inverted Meckel's diverticulum. PMID- 21621495 TI - Myth: mechanical ventilation is a therapeutic relic. AB - Non-invasive respiratory support techniques such as continuous positive airway pressure (CPAP) have been increasingly used for management of surfactant deficient lung disease in preterm infants. The successful use of this approach depends upon the condition of the baby at birth and requires the establishment of spontaneous breathing at birth. The reported advantages of CPAP in observational studies demonstrating a reduction in chronic lung disease have not been substantiated in recently reported well-designed randomised trials. This approach is now more established in larger and more mature preterm infants, and proper patient selection with close observation should be exercised when used in extremely low gestational age infants. PMID- 21621496 TI - Tuberculous dactylitis in the setting of low serum vitamin D: a case report. AB - We present the case of a previously well patient who presented to the Emergency Department of a Dublin hospital with a tuberculous infection of his dominant index finger and a very low serum vitamin D level--this has been implicated in both primary and reactivation infections with Mycobacterium Tuberculosis. This case highlights and reviews both the importance of considering non-endemic pathologies in the setting of a patient base of diverse ethnicity, and the emerging importance of vitamin D in the immune response to M. tuberculosis infection. We discuss the relevant literature to highlight the background of this disease process, and the importance of a multidisciplinary approach to these patients. PMID- 21621497 TI - Detection of methylated tissue factor pathway inhibitor 2 and human long DNA in fecal samples of patients with colorectal cancer in China. AB - BACKGROUND: To investigate the feasibility of detecting methylated tissue factor pathway inhibitor (TFPI2) and quantifying human long DNA with fluorescent quantitative Alu PCR in fecal DNA as a non-invasive screening tool for colorectal cancer (CRC). MATERIALS AND METHODS: Methylation-specific PCR (MSP) was performed to analyze TFPI2 gene promoter methylation status in a blinded fashion in stool samples taken from 30 endoscopically diagnosed healthy controls, 20 patients with adenomas, and 60 patients with colorectal cancer. Real-time Alu PCR was used to quantify human long DNA. RESULTS: The specificity of fecal TFPI2 MSP assay and long DNA assay was 100% and 83.3%, respectively. The sensitivity of fecal TFPI2 MSP assay and long DNA assay was 68.3% and 53.3%, respectively. The sensitivity of fecal DNA assay (either marker being positive) was 86.7%, which was high for CRC. CONCLUSIONS: Our results have demonstrated the feasibility of using TFPI2 methylation and quantify human long DNA with fluorescent quantitative Alu PCR in fecal samples as a new noninvasive test for CRC. PMID- 21621498 TI - Trends in invasive breast cancer incidence among French women not exposed to organized mammography screening: an age-period-cohort analysis. AB - BACKGROUND: The long tenure of the Doubs cancer registry (France) and the late implementation of a mass screening program provide a unique opportunity to assess the relative contributions of age, period and cohort effects to the increase in female invasive breast cancer incidence, while avoiding the influence of an organized screening program. METHODS: Population and incidence data were provided for the Doubs region during the 1978-2003 period. Breast cancer counts and person years were tabulated into 1-year classes by age and time period. Age-period cohort models with parametric smooth functions were fitted to the data, assuming a Poisson distribution for the number of observed cases. RESULTS: A total of 5688 incident cases of invasive breast cancer in women were diagnosed in women aged 30 84 years in the Doubs region between 1978 and 2003. The annual percentage increase in incidence is 2.09%. Age effects rise dramatically until age 50, and at a slower pace afterwards. Large cohort curvature effects (p<10(-6)), show departure from linear trends, with a significant peak for women born around 1940. Period curvature effects are lower in magnitude (p=0.01). CONCLUSION: Both cohort and period effects are involved in the marked increase in breast cancer incidence over a 25-year period in the Doubs region. Although the future trend for breast cancer incidence is difficult to predict, the introduction of an organized screening program, and the sharp decline in hormone replacement therapy use will likely contribute to period effects in future analyses. PMID- 21621499 TI - Assessment of follow-up, and the completeness and accuracy of cancer case ascertainment in three areas of India. AB - BACKGROUND: A prospective study of diet and cancer has not been conducted in India; consequently, little is known regarding follow-up rates or the completeness and accuracy of cancer case ascertainment. METHODS: We assessed follow-up in the India Health Study (IHS; 4671 participants aged 35-69 residing in New Delhi, Mumbai, or Trivandrum). We evaluated the impact of medical care access and relocation, re-contacted the IHS participants to estimate follow-up rates, and conducted separate studies of cancer cases to evaluate registry coverage (604 cases in Trivandrum) and the accuracy of self- and proxy-reporting (1600 cases in New Delhi and Trivandrum). RESULTS: Over 97% of people reported seeing a doctor and 85% had lived in their current residence for over six years. The 2-year follow-up rate was 91% for Trivandrum and 53% for New Delhi. No cancer cases were missed among public institutions participating in the surveillance program in Trivandrum during 2003-2004; but there are likely to be unmatched cases (ranging from 5 to 13% of total cases) from private hospitals in the Trivandrum registry, as there are no mandatory reporting requirements. Vital status was obtained for 36% of cancer cases in New Delhi as compared to 78% in Trivandrum after a period of 4 years. CONCLUSIONS: A prospective cohort study of cancer may be feasible in some centers in India with active follow-up to supplement registry data. Inclusion of cancers diagnosed at private institutions, unique identifiers for individuals, and computerized medical information would likely improve cancer registries. PMID- 21621500 TI - Colorimetric inorganic pyrophosphate assay using a double cycling enzymatic method. AB - Pyruvate phosphate dikinase (PPDK, EC 2.7.9.1) from the hyperthermophile Thermotoga maritima was biochemically characterized with the aim of establishing a colorimetric assay for inorganic pyrophosphate (PPi). When heterologously expressed in Escherichia coli, T. maritima PPDK (TmPPDK) was far more stable any other PPDK reported so far: it retained >90% of its activity after incubation for 1 h at 80 degrees C, and >80% of its activity after incubation for 20 min at pHs ranging from 6.5 to 10.5 (50 degrees C). In contrast to PPDKs from protozoa and plants, this TmPPDK showed very long-term stability at low temperature: full activity was retained even after storage for at least 2 years at 4 degrees C. TmPPDK was successfully applied to a novel colorimetric PPi assay, which employed (i) a PPi cycling reaction using TmPPDK and nicotinamide mononucleotide adenylyltransferase (EC 2.7.7.1) from Saccharomyces cerevisiae and (ii) a NAD cycling reaction to accumulate reduced nitroblue tetrazolium (diformazan). This enabled detection of 0.2 MUM PPi, making this method applicable for preliminary measurement of PPi levels in PCR products in an automatic clinical analyzer. PMID- 21621501 TI - Interactions among the SNARE proteins and complexin analyzed by a yeast four hybrid assay. AB - Exocytosis is one of the most crucial and ubiquitous processes in all of biology. This event is mediated by the formation of SNARE (soluble N-ethylmaleimide sensitive factor attachment protein receptor) complexes, ternary assemblies of syntaxin, SNAP23/SNAP25 (synaptosomal-associated protein of 23 or 25 kDa), and synaptobrevin. The exocytotic process can be further regulated by complexin, which interacts with the SNARE complex. Complexin is involved in a Ca(2+) triggered exocytotic process. In eukaryotic cells, multiple isoforms of SNARE proteins are expressed and are involved in distinct types of exocytosis. To understand the underlying biochemical mechanism of various exocytotic processes mediated by different SNARE protein isoforms, we systematically analyzed the interactions among syntaxin, SNAP23/SNAP25, synaptobrevin, and complexin by employing a newly developed yeast four-hybrid interaction assay. The efficiency of SNARE complex formation and the specificity of complexin binding are regulated by the different SNARE protein isoforms. Therefore, various types of exocytosis, occurring on different time scales with different efficiencies, can be explained by the involved SNARE complexes composed of different combinations of SNARE protein isoforms. PMID- 21621502 TI - A continuous fluorescence displacement assay for BioA: an enzyme involved in biotin biosynthesis. AB - Cofactor biosynthetic pathways represent a rich source of potential antibiotic targets. The second step in biotin biosynthesis is performed by BioA, a pyridoxal 5'-phosphate (PLP)-dependent enzyme. This enzyme has been confirmed as a candidate target in Mycobacterium tuberculosis; however, the current bioassay used to measure BioA activity is cumbersome and low throughput. Here we describe the design, development, and optimization of a continuous coupled fluorescence displacement assay to measure BioA activity. In this coupled assay, BioD converts the product of the BioA-catalyzed reaction into dethiobiotin, which is subsequently detected by displacement of a fluorescently labeled dethiobiotin probe from streptavidin. The assay was further adapted to a high-throughput screening format and validated against the LOPAC(1280) library. PMID- 21621503 TI - Role of cytosolic phospholipase A(2)alpha in cell rounding and cytotoxicity induced by ceramide-1-phosphate via ceramide kinase. AB - Ceramide-1-phosphate (C1P), produced by ceramide kinase (CERK), is implicated in the regulation of many biological functions including cell growth and inflammation. C1P is a direct activator of group IVA cytosolic phospholipsase A(2) (PLA2G4A or cPLA(2)alpha). Although activation of the CERK-C1P pathway causes mitogenic and cytoprotective responses in many cells, the pathway shows cytotoxicity in several cells and the precise mechanism has not been elucidated. In the present study, we examined the effect of human CERK (hCERK) expression on cytotoxicity in two cell lines. Expression of hCERK in CHO cells caused cell rounding and lactate dehydrogenase (LDH) leakage, and co-addition of ceramide enhanced these responses. Expression of hCERK enhanced C1P formation and release of arachidonic acid in Ca(2+) ionophore-stimulated cells. Treatment with 20MUM C2 C1P for 24 h caused cell rounding, and the response was significantly decreased by an inhibitor of cPLA(2)alpha. In L929 cells, expression of hCERK with and without ceramide caused cell rounding and LDH leakage, respectively, and the responses were significantly less in a stable clone of L929 cells lacking cPLA(2)alpha. These findings suggest the involvement of cPLA(2)alpha in CERK-C1P pathway-induced cytotoxicity. PMID- 21621504 TI - Upregulation of the mitochondrial transport protein, Tim50, by mutant p53 contributes to cell growth and chemoresistance. AB - The p53 gene is one of the most frequently mutated genes in human cancer. Some p53 mutations impart additional functions that promote oncogenesis. To investigate how these p53 mutants function, a proteomic analysis was performed. The protein, translocator of the inner mitochondrial membrane 50 (Tim50), was upregulated in a non-small cell lung carcinoma cell line (H1299) that expressed the p53 mutants R175H and R273H compared to cells lacking p53. Tim50 was also elevated in the breast cancer cell lines MDA-MB-468 and SK-BR-3, that endogenously express the p53 mutants R175H and R273H, respectively, compared to MCF-10A. The p53 mutants R175H and R273H, but not WT p53, upregulated the expression of a Tim50 promoter construct and chromatin immunoprecipitation (ChIP) analysis indicated increased histone acetylation and increased interaction of the transcription factors Ets-1, CREB and CREB-binding protein (CBP) with the Tim50 promoter in the presence of mutant p53. Finally, reduction of Tim50 expression reduced the growth rate and chemoresistance of cells harboring mutant p53 but had no effect upon cells lacking p53. Taken together, these findings identify the Tim50 gene as a transcriptional target of mutant p53 and suggest a novel mechanism by which p53 mutants enhance cell growth and chemoresistance. PMID- 21621505 TI - Specific antioxidant selenoproteins are induced in the heart during hypertrophy. AB - Selenium (Se) is thought to confer cardioprotective effects through the actions of antioxidant selenoprotein enzymes that directly limit levels of ROS such as hydrogen peroxide (H(2)O(2)) or that reverse oxidative damage to lipids and proteins. To determine how the selenoproteome responds to myocardial hypertrophy, two mouse models were employed: triidothyronine (T3)- or isoproterenol (ISO) treatment. After 7days of T3- and ISO-treatment, cardiac stress was demonstrated by increased H(2)O(2) and caspase-3 activity. Neither treatment produced significant increases in phospholipid peroxidation or TUNEL-positive cells, suggesting that antioxidant systems were protecting the cardiomyocytes from damage. Many selenoprotein mRNAs were induced by T3- and ISO-treatment, with levels of methionine sulfoxide reductase 1 (MsrB1, also called SelR) mRNA showing the largest increases. MsrB enzymatic activity was also elevated in both models of cardiac stress, while glutathione peroxidase (GPx) activity and thioredoxin reductase (Trxrd) activity were moderately and nonsignificantly increased, respectively. Western blot assays revealed a marked increase in MsrB1 and moderate increases in GPx3, GPx4, and Trxrd1, particularly in T3-treated hearts. Thus, the main response of the selenoproteome during hypertrophy does not involve increased GPx1, but increased GPx3 for reducing extracellular H(2)O(2) and increased GPx4, Trxrd1, and MsrB1 for minimizing intracellular oxidative damage. PMID- 21621506 TI - Biochemical and biophysical characterisation yields insights into the mechanism of a Cd/Zn transporting ATPase purified from the hyperaccumulator plant Thlaspi caerulescens. AB - TcHMA4 (GenBank no. AJ567384), a Cd/Zn transporting ATPase of the P(1B)-type (=CPx-type) was isolated and purified from roots of the Cd/Zn hyperaccumulator Thlaspi caerulescens. Optimisation of the purification protocol, based on binding of the natural C-terminal His-tag of the protein to a Ni-IDA metal affinity column, yielded pure, active TcHMA4 in quantities sufficient for its biochemical and biophysical characterisation with various techniques. TcHMA4 showed activity with Cu(2+), Zn(2+) and Cd(2+) under various concentrations (tested from 30nM to 10MUM), and all three metal ions activated the ATPase at a concentration of 0.3MUM. Notably, the enzyme worked best at rather high temperatures, with an activity optimum at 42 degrees C. Arrhenius plots yielded interesting differences in activation energy. In the presence of zinc it remained constant (E(A)=38kJ?mol(-1)) over the whole concentration range while it increased from 17 to 42kJ?mol(-1) with rising copper concentration and decreased from 39 to 23kJ?mol(-1) with rising cadmium concentration. According to EXAFS the TcHMA4 appeared to bind Cd(2+) mainly by thiolate sulphur from cysteine, and not by imidazole nitrogen from histidine. PMID- 21621507 TI - The behavior of sea anemone actinoporins at the water-membrane interface. AB - Actinoporins constitute a group of small and basic alpha-pore forming toxins produced by sea anemones. They display high sequence identity and appear as multigene families. They show a singular behaviour at the water-membrane interface: In aqueous solution, actinoporins remain stably folded but, upon interaction with lipid bilayers, become integral membrane structures. These membranes contain sphingomyelin, display phase coexistence, or both. The water soluble structures of the actinoporins equinatoxin II (EqtII) and sticholysin II (StnII) are known in detail. The crystalline structure of a fragaceatoxin C (FraC) nonamer has been also determined. The three proteins fold as a beta sandwich motif flanked by two alpha-helices, one of them at the N-terminal end. Four regions seem to be especially important: A cluster of aromatic residues, a phosphocholine binding site, an array of basic amino acids, and the N-terminal alpha-helix. Initial binding of the soluble monomers to the membrane is accomplished by the cluster of aromatic amino acids, the array of basic residues, and the phosphocholine binding site. Then, the N-terminal alpha-helix detaches from the beta-sandwich, extends, and lies parallel to the membrane. Simultaneously, oligomerization occurs. Finally, the extended N-terminal alpha helix penetrates the membrane to build a toroidal pore. This model has been however recently challenged by the cryo-EM reconstruction of FraC bound to phospholipid vesicles. Actinoporins structural fold appears across all eukaryotic kingdoms in other functionally unrelated proteins. Many of these proteins neither bind to lipid membranes nor induce cell lysis. Finally, studies focusing on the therapeutic potential of actinoporins also abound. PMID- 21621508 TI - The B degrees AT1 amino acid transporter from rat kidney reconstituted in liposomes: kinetics and inactivation by methylmercury. AB - The neutral amino acid transporter B degrees -like from rat kidney, previously reconstituted in liposomes, was identified as B degrees AT1 by a specific antibody. Collectrin was present in the brush-border extract but not in functionally active proteoliposomes, indicating that it was not required for the transport function. Neutral amino acids behaved as competitive inhibitors of the glutamine transport mediated by B degrees AT1 with half saturation constants ranging from 0.13 to 4.74mM. The intraliposomal half saturation constant for glutamine was 2.0mM. By a bisubstrate kinetic analysis of the glutamine-Na(+) cotransport, a random simultaneous mechanism was found. Methylmercury and HgCl(2) inhibited the transporter; the inhibition was reversed by dithioerythritol, Cys and, at a lower extent, N-acetylcysteine but not by S-carboxymethylcysteine. The IC(50) of the transporter for methylmercury and HgCl(2) was 1.88 and 1.75MUM, respectively. The reagents behaved as non-competitive inhibitors toward both glutamine and Na(+) and no protection by glutamine or Na(+) was found for the two inhibitors. PMID- 21621509 TI - The glucose transporter-2 (GLUT2) is a low affinity dehydroascorbic acid transporter. AB - We studied the acquisition of dehydroascorbic acid by rat hepatocytes, H4IIE rat hepatoma cells and Xenopus laevis oocytes. Transport kinetics and competition and inhibition studies revealed that rat hepatocytes transport oxidized dehydroascorbic acid through a single functional component possessing the functional and kinetic properties expected for the glucose transporter GLUT2. On the other hand, rat hepatoma cells showed expression of at least two dehydroascorbic acid transporters with the expected functional and kinetic properties expected for GLUT1 and GLUT2. Expression studies of GLUT2 in X. laevis oocytes followed by transport kinetics and competition and inhibition studies revealed that GLUT2 is a low affinity dehydroascorbic transporter whose kinetic and functional properties match those observed for the endogenous GLUT2 transporter in rat hepatocytes and rat hepatoma cells. Therefore, GLUT2, a transporter known as a low affinity transporter of glucose and fructose and a high affinity transporter of glucosamine is also a low affinity dehydroascorbic acid transporter. PMID- 21621510 TI - Fgf20b is required for the ectomesenchymal fate establishment of cranial neural crest cells in zebrafish. AB - In cranial skeletal development, the establishment of the ectomesenchymal lineage within the cranial neural crest is of great significance. Fgfs are polypeptide growth factors with diverse functions in development and metabolism. Fgf20b knockdown zebrafish embryos showed dysplastic neurocranial and pharyngeal cartilages. Ectomesenchymal cells from cranial neural crest cells were significantly decreased in Fgf20b knockdown embryos, but cranial neural crest cells with a non-ectomesnchymal fate were increased. However, the proliferation and apoptosis of cranial neural crest cells were essentially unchanged. Fgfr1 knockdown embryos also showed dysplastic neurocranial and pharyngeal cartilages. The present findings indicate that Fgf20b is required for ectomesenchymal fate establishment via the activation of Fgfr1 in zebrafish. PMID- 21621511 TI - Fucoxanthin exerts differing effects on 3T3-L1 cells according to differentiation stage and inhibits glucose uptake in mature adipocytes. AB - Progression of 3T3-L1 preadipocyte differentiation is divided into early (days 0 2, D0-D2), intermediate (days 2-4, D2-D4), and late stages (day 4 onwards, D4-). In this study, we investigated the effects of fucoxanthin, isolated from the edible brown seaweed Petalonia binghamiae, on adipogenesis during the three differentiation stages of 3T3-L1 preadipocytes. When fucoxanthin was applied during the early stage of differentiation (D0-D2), it promoted 3T3-L1 adipocyte differentiation, as evidenced by increased triglyceride accumulation. At the molecular level, fucoxanthin increased protein expression of peroxisome proliferator-activated receptor gamma (PPARgamma), CCAAT/enhancer-binding protein alpha (C/EBPalpha), sterol regulatory element-binding protein 1c (SREBP1c), and aP2, and adiponectin mRNA expression, in a dose-dependent manner. However, it reduced the expression of PPARgamma, C/EBPalpha, and SREBP1c during the intermediate (D2-D4) and late stages (D4-D7) of differentiation. It also inhibited the uptake of glucose in mature 3T3-L1 adipocytes by reducing the phosphorylation of insulin receptor substrate 1 (IRS-1). These results suggest that fucoxanthin exerts differing effects on 3T3-L1 cells of different differentiation stages and inhibits glucose uptake in mature adipocytes. PMID- 21621512 TI - Nitric oxide counters the inhibitory effects of uremic toxin indoxyl sulfate on endothelial cells by governing ERK MAP kinase and myosin light chain activation. AB - Uremic toxins such as indoxyl sulfate (IS) accumulate at a high level in end stage renal disease (ESRD) and can exhibit significant systemic endothelial toxicity leading to accelerated cardiovascular events. The precise molecular mechanisms by which IS causes endothelial dysfunction are unknown. We tested the hypothesis that IS negatively influences properties of endothelial cells, such as migration and tube formation, by depleting nitric oxide (NO) bioavailability, and that an NO donor can reverse these inhibitory effects. IS inhibited human umbilical vein endothelial cell (HUVEC) migration and formation of tubes on matrigel. Mechanistically, IS inhibited VEGF-induced NO release from HUVECs. An NO donor, SNAP, reversed IS-mediated inhibition of HUVEC migration as well as tube-formation. IS inhibited ERK 1/2 MAP kinase activity in a dose-dependent manner, but this was preserved by SNAP. Inhibition of ERK 1/2 with a pharmacological inhibitor (U0126) decreased HUVEC migration and tube formation; these effects too were prevented by SNAP. Further, IS stimulated activation of myosin light chain (MLC), potentially stimulating endothelial contractility, while SNAP decreased MLC activation. Thus, we conclude that the negative effects of IS on endothelial cells are prevented, to a major extent, by NO, via its divergent actions on ERK MAP kinase and MLC. PMID- 21621513 TI - Casuarinin suppresses TNF-alpha-induced ICAM-1 expression via blockade of NF kappaB activation in HaCaT cells. AB - Hippophae rhamnoides has been extensively used in oriental traditional medicines for treatment of asthma, skin diseases, gastric ulcers, and lung disorders. In this study, we isolated casuarinin from the leaves of H.rhamnoides and examined the effect of casuarinin on the TNF-alpha-induced ICAM-1 expression in a human keratinocytes cell line HaCaT. Pretreatment with casuarinin inhibited TNF-alpha induced protein and mRNA expression of ICAM-1 and subsequent monocyte adhesiveness in HaCaT cells. Casuarinin significantly inhibited TNF-alpha-induced NF-kappaB activation. In addition, casuarinin inhibited activation of ERK and p38 MAPK in a dose-dependent manner. Furthermore, pretreatment with casuarinin decreased TNF-alpha-induced pro-inflammatory mediators, such as IL-1beta, IL-6, IL-8, and MCP-1. These results demonstrated that casuarinin exerts its anti inflammatory activity by suppressing TNF-alpha-induced expression of ICAM-1 and pro-inflammatory cytokines/chemokines via blockage of activation of NF-kappaB and ERK/p38 MAPK and can be used as a therapeutic agent against inflammatory skin diseases. PMID- 21621514 TI - Lipid accumulation and lysosomal pathways contribute to dysfunction and apoptosis of human endothelial cells caused by 7-oxysterols. AB - Endothelial dysfunction and cell death play an important role in pathogenesis of atherosclerosis. 7-Oxysterols, the major cytotoxic component found in oxidized low-density lipoprotein, are toxic to endothelial cells. However, the pathways and molecular mechanism involved in the process remain incompletely understood. In this study, we first investigate whether 7beta-hydroxycholesterol (7betaOH) or 7-ketocholesterol (7keto) induces apoptosis of human endothelial cell line (HUVEC CS). We then examine possible involved pathways by focusing on cellular lipid, lysosomal pathways, cellular oxidative stress and mitochondrial pathways. Our results for the first time showed that 7-oxysterols induced apoptotic cell death of HUVEC-CS after 24h, which was preceded by early lipid accumulation (6h) and lysosomal membrane permeabilization (6-12h). Afterward, levels of reactive oxygen species, mitochondrial membrane permeabilization, and lysosomal cathepsin were increased assayed by immuno-cytochemistry and blotting. Notably, the exposure to 7betaOH or 7keto induced expressions and secretion of isoforms of von Willebrand factor (VWF). We conclude that apoptosis of HUVEC-CS induced by 7betaOH or 7keto mediates by early lysosomal lipid accumulation and oxidative lysosomal pathways, which results in induction and release of VWF. The results suggest the cell death induced by 7-oxysterols may contribute to endothelial dysfunction and atherothrombosis. PMID- 21621515 TI - Genes that integrate multiple adipogenic signaling pathways in human mesenchymal stem cells. AB - Adipogenesis is a well-characterized cell differentiation process. A large body of evidence has revealed the core transcription factors and signaling pathways that govern adipogenesis, but cross-talks between these cellular signals and its functional consequences have not been thoroughly investigated. We, therefore, sought to identify genes that are regulated by multiple signaling pathways during adipogenesis of human mesenchymal stem cells. Focusing on the early stage of adipogenesis, microarray analysis and quantitative RT-PCR identified 12 genes whose transcription levels were dramatically affected by the complete adipogenic induction cocktail but not by the cocktail's individual components. Expression kinetics of these genes indicate diverse mechanisms of transcriptional regulation during adipogenesis. Functional relationships between these genes and adipogenic differentiation were frequently unknown. This study thus provided novel adipogenic gene candidates that likely mediate communications among multiple signaling pathways within human mesenchymal stem cells. PMID- 21621516 TI - Paradoxical effects of chondroitin sulfate-E on Japanese encephalitis viral infection. AB - Glycosaminoglycans (GAGs) have diverse functions in the body and are involved in viral infection. The purpose of this study was to evaluate the possible roles of the E-disaccharide units GlcAbeta1-3GalNAc(4,6-O-disulfate) of chondroitin sulfate (CS), a GAG involved in neuritogenesis and neuronal migration, in Japanese encephalitis virus (JEV) infection. Soluble CS-E (sCS-E) derived from squid cartilage inhibited JEV infection in African green monkey kidney-derived Vero cells and baby hamster kidney-derived BHK cells by interfering with viral attachment. In contrast, sCS-E enhanced viral infection in the mouse neuroblastoma cell line Neuro-2a, despite the fact that viral attachment to Neuro 2a cells was inhibited by sCS-E. This enhancement effect in Neuro-2a cells seemed to be related to increased viral RNA replication and was also observed in a rat infection model in which intracerebral coadministration of sCS-E with JEV in 17 day-old rats resulted in higher brain viral loads than in rats infected without sCS-E administration. These results show the paradoxical effects of sCS-E on JEV infection in different cell types and indicate that potential use of sCS-E as an antiviral agent against JEV infection should be approached with caution considering its effects in the neuron, the major target of JEV. PMID- 21621517 TI - IGF-1Ea induces vessel formation after injury and mediates bone marrow and heart cross-talk through the expression of specific cytokines. AB - The aim of this study was to investigate whether supplemental IGF-1Ea transgene expression induces activation of local cardiac and bone marrow stem cell population to mediate mammalian heart repair. In physiologic conditions, cardiac overexpression of the IGF-1Ea propeptide is associated with an enrichment of c Kit/Sca-1 positive side population cells in the bone marrow and the occurrence of an endothelial-primed CD34 positive side population in the heart. This cellular profile is shown here to correlate with the expression of cytokines involved in stem cell mobilization and vessel formation. This molecular and cellular interplay favored IGF-1Ea-mediated vessel formation in injured hearts. The physiologic and pathologic connection between cytokines and stem cells in response to IGF-1Ea may represent an important model to understand how to elicit endogenous reparative signaling. PMID- 21621518 TI - Electrogenic Na+/HCO3- co-transporter-1 is essential for the parathyroid hormone stimulated intestinal HCO3- secretion. AB - Parathyroid hormone (PTH) was recently demonstrated to enhance the HCO(3)(-) secretion through the apical anion channel, cystic fibrosis transmembrane conductance regulator (CFTR), but how the HCO(3)(-) entered the epithelial cells was not well understood, in part, due to the lack of specific inhibitors of the basolateral HCO(3)(-) transporters. Moreover, the function of the PTH-stimulated HCO(3)(-) secretion has never been investigated in vivo. Here, we designed three specific pairs of small interfering RNA sequences to simultaneously knockdown three variants of the electrogenic Na(+)/HCO(3)(-) co-transporter (NBCe)-1 in the intestinal epithelium-like Caco-2 monolayer. The results showed that NBCe1 mRNA levels were markedly reduced, and the PTH-induced transepithelial current and voltage changes were diminished after triple knockdown as determined by quantitative real-time PCR and Ussing chamber technique, respectively. An in vivo ligated intestinal loop study further showed that there was an increased fluid secretion, presumably driven by HCO(3)(-) transport, in the ileum, but not in jejunum or colon, of rats administered intravenously with 2 MUg/kg body weight of rat PTH 1-34. Therefore, the present results suggested that PTH stimulated intestinal HCO(3)(-) secretion, particularly in the ileum, by inducing the basolateral HCO(3)(-) uptake via NBCe1. PMID- 21621519 TI - Roles of ATR1 paralogs YMR279c and YOR378w in boron stress tolerance. AB - Boron is a necessary nutrient for plants and animals, however excess of it causes toxicity. Previously, Atr1 and Arabidopsis Bor1 homolog were identified as the boron efflux pump in yeast, which lower the cytosolic boron concentration and help cells to survive in the presence of toxic amount of boron. In this study, we analyzed ATR1 paralogs, YMR279c and YOR378w, to understand whether they participate in boron stress tolerance in yeast. Even though these genes share homology with ATR1, neither their deletion rendered cells boron sensitive nor their expression was significantly upregulated by boron treatment. However, expression of YMR279, but not YOR378w, from the constitutive GAPDH promoter on a high copy plasmid provided remarkable boron resistance by decreasing intracellular boron levels. Thus our results suggest the presence of a third boron exporter, YMR279c, which functions similar to ATR1 and provides boron resistance in yeast. PMID- 21621520 TI - Lack of CaV3.1 channels causes severe motor coordination defects and an age dependent cerebellar atrophy in a genetic model of essential tremor. AB - T-type Ca(2+) channels have been implicated in tremorogenesis and motor coordination. The alpha1 subunit of the Ca(V)3.1 T-type Ca(2+) channel is highly expressed in motor pathways in the brain, but knockout of the Ca(V)3.1 gene (alpha(1G)(-/-)) per se causes no motor defects in mice. Thus, the role of Ca(V)3.1 channels in motor control remains obscure in vivo. Here, we investigated the effect of the Ca(V)3.1 knockout in the null genetic background of alpha1 GABA(A) receptor (alpha1(-/-)) by generating the double mutants (alpha1(-/ )/alpha(1G)(-/-)). alpha1(-/-)/alpha(1G)(-/-) mice showed severer motor abnormalities than alpha1(-/-) mice as measured by potentiated tremor activities at 20Hz and impaired motor learning. Propranolol, an anti-ET drug that is known to reduce the pathologic tremor in alpha1(-/-) mice, was not effective for suppressing the potentiated tremor in alpha1(-/-)/alpha(1G)(-/-) mice. In addition, alpha1(-/-)/alpha(1G)(-/-) mice showed an age-dependent loss of cerebellar Purkinje neurons. These results suggest that alpha1(-/-)/alpha(1G)(-/ ) mice are a novel mouse model for a distinct subtype of ET in human and that Ca(V)3.1 T-type Ca(2+) channels play a role in motor coordination under pathological conditions. PMID- 21621521 TI - Agonist-induced formation of FGFR1 homodimers and signaling differ among members of the FGF family. AB - Fibroblast growth factor receptor 1 (FGFR1) is known to be activated by homodimerization in the presence of both the FGF agonist ligand and heparan sulfate glycosaminoglycan. FGFR1 homodimers in turn trigger a variety of downstream signaling cascades via autophosphorylation of tyrosine residues in the cytoplasmic domain of FGFR1. By means of Bioluminescence Energy Resonance Transfer (BRET) as a sign of FGFR1 homodimerization, we evaluated in HEK293T cells the effects of all known FGF agonist ligands on homodimer formation. A significant correlation between BRET(2) signaling and ERK1/2 phosphorylation was observed, leading to a further characterization of the binding and signaling properties of the FGF subfamilies. FGF agonist ligand-FGFR1 binding interactions appear as the main mechanism for the control of FGFR1 homodimerization and MAPK signaling which demonstrated a high correlation. The bioinformatic analysis demonstrates the interface of the two pro-triplets SSS (Ser-Ser-Ser) and YGS (Tyr Gly-Ser) located in the extracellular and intracellular domain of the FGFR1. These pro-triplets are postulated participate in the FGFR1 homodimerization interface interaction. The findings also reveal that FGF agonist ligands within the same subfamily of the FGF gene family produced similar increases in FGFR1 homodimer formation and MAPK signaling. Thus, the evolutionary relationship within this gene family appears to have a distinct functional relevance. PMID- 21621522 TI - ZL11n is a novel nitric oxide-releasing derivative of farnesylthiosalicylic acid that induces apoptosis in human hepatoma HepG2 cells via MAPK/mitochondrial pathways. AB - ZL11n is a novel furoxan-based nitric oxide (NO)-releasing derivative of farnesylthiosalicylic acid. In this study, we examined the anticancer effects and the potential mechanism of action of ZL11n in vitro and in vivo. It was found that ZL11n exhibited a favorable, selective cytotoxic effect in the HepG2 cell line. The yield of NO in the ZL11n treated HepG2 cells was much higher than in the control group and the normal human liver L-02 cells. Furthermore, the NO concentration was correlated to the degree of cytotoxicity observed. The ZL11n induced apoptosis was assessed by Annexin V-FITC/propidium iodide flow cytometry assay. ZL11n triggered the mitochondrial/caspase apoptotic pathway by decreasing mitochondrial membrane potential, cytochrome c release from mitochondrial, and reducing the Bcl-2-to-Bax ratio, in addition to activating the caspase cascade. Simultaneously, we found that ZL11n treatment led to an increase in JNK and ERK1/2 phosphorylation. Furthermore, treatment with SP600125 (a JNK inhibitor) and PD98059 (an ERK1/2 inhibitor) prior to ZL11n treatment was found to significantly reverse ZL11n-induced apoptosis. The in vivo findings also revealed that ZL11n significantly reduced tumor volume and weight in the H(22) solid tumor mouse model examined. In short, our findings suggest that ZL11n induced apoptosis through the coordination of the mitochondrial apoptotic pathway (activated by NO) and MAPKs signaling pathway (triggered by JNK or ERK). PMID- 21621523 TI - Human papillomavirus 16E6 suppresses major histocompatibility complex class I by upregulating lymphotoxin expression in human cervical cancer cells. AB - Major histocompatibility complex (MHC) class I is a major host defense mechanism against viral infections such as type 16 and type 18 of the human papillomavirus (HPV). Here, we found that the E6 oncogene from HPV16, but not HPV18, suppressed MHC I expression. Ectopic expression of HPV16E6 in HeLa cells, which are infected with HPV18, suppressed MHC I expression, and that knockdown by antisense or siRNA of the HPV16E6 strongly enhanced MHC I expression in Caski cells, which are infected with HPV18, but not HPV16. The expression of HPV16E6 strongly enhanced cellular resistance to cytotoxic T lymphocytes (CTLs)-mediated lytic activity, and knockdown of HPV16E6 by antisense had the opposite effect. The regulation of HPV16E6-mediated MHC I suppression might be through the regulation of lymphotoxin (LT) and its receptor, LTbetaR. In addition, cells from the spleen and liver of LTalpha- or LTbetaR-deficient mice showed increased MHC I expression. Overall, these results demonstrated that the E6 oncogene of HPV16 might play an important role in cell transformation and cancer development through LT-mediated MHC I downregulation in humans. PMID- 21621524 TI - Protein cross-linking as a novel mechanism of action of a ubiquitin-activating enzyme inhibitor with anti-tumor activity. AB - Ubiquitin-activating enzyme 1 (UBE1) is a critical regulator of the ubiquitination cycle and its targeted inhibition may be an appropriate therapeutic strategy as tumor cells are reported to have increased dependence on protein ubiquitination. PYR-41 is a small molecule with previously described UBE1 inhibitory activity. PYR-41 blocks ubiquitination reactions but paradoxically leads to the accumulation of high MW ubiquitinated proteins. Detailed evaluation of PYR-41 activity demonstrated that PYR-41 inhibited UBE1 activity but also had equal or greater inhibitory activity against several deubiquitinases (DUBs) in intact cells and purified USP5 in vitro. Both UBE1 and DUB inhibition were mediated through PYR-41-induced covalent protein cross-linking which paralleled the inhibition of the target proteins enzymatic activity. PYR-41 also mediated cross-linking of specific protein kinases (Bcr-Abl, Jak2) to inhibit their signaling activity. Chemical reactivity modeling provided some insight into the cross-linking potential and partial target selectivity of PYR-41. Overall, our results suggest a broader range of targets and a novel mechanism of action for this UBE1 inhibitor. In addition, since PYR-41-related compounds have demonstrated anti-tumor activity in animal studies, partially selective protein cross-linking may represent an alternate approach to affect signal transduction modules and ubiquitin cycle-regulatory proteins for cancer therapy. PMID- 21621525 TI - Glabridin protects osteoblastic MC3T3-E1 cells against antimycin A induced cytotoxicity. AB - Mitochondrial dysfunction, particularly respiratory chain disruption, is often responsible for aging-related bone diseases. In this study, the protective effects of glabridin, an isoflavan isolated from licorice root, against pharmacological inhibition of the respiratory chain were studied using osteoblastic MC3T3-E1 cells treated with antimycin A, which inhibits complex III of the electron transport system. Glabridin restored mitochondrial membrane potential dissipation, ATP loss, inactivation of complex IV, intracellular calcium elevation, and cytochrome c release that was induced by antimycin A treatment. This compound also prevented cell death. These results imply that glabridin protects osteoblasts from antimycin A-induced cell death via improved mitochondrial function. Glabridin scavenged ROS and mitochondrial superoxide anions generated by antimycin A. In addition, glabridin prevented antimycin A induced nitrotyrosine increase and thioredoxin reductase inactivation, suggesting that glabridin may be useful for protecting mitochondria against a burst of oxidative stress. Since phosphoinositide 3-kinase (PI3K) and cAMP-response element-binding protein (CREB) signaling is known to be pro-survival, we determined whether PI3K and CREB activation is associated with the cytoprotective effects of glabridin in the MC3T3-E1 cells. Glabridin restored antimycin A induced inactivation of PI3K and CREB, suggesting that PI3K and CREB-dependent pathways may be involved in glabridin-induced cytoprotective responses. Our study demonstrates that glabridin reduces mitochondrial dysfunction induced during aging, and could significantly prevent osteoblast damage in osteoporotic patients. PMID- 21621526 TI - Nitric oxide donor, V-PROLI/NO, provides protection against arsenical induced toxicity in rat liver cells: requirement for Cyp1a1. AB - Arsenic is a cancer chemotherapeutic but hepatotoxicity can be a limiting side effect. O(2)-vinyl 1-[2-(carboxylato)pyrrolidin-1-yl]diazen-1-ium-1,2-diolate (V PROLI/NO) is a nitric oxide (NO) donor prodrug and metabolized by liver cytochromes P450 (CYP450) to release NO. The effects of V-PROLI/NO pretreatment on the toxicity of arsenic (as NaAsO(2)) were studied in a rat liver cell line (TRL 1215). The cells acted upon the prodrug to release NO, as assessed by nitrite levels, in a time-dependent fashion to maximal levels of 8-fold above basal levels. Pretreatment with V-PROLI/NO markedly reduced arsenic cytolethality which was directly related to the level of NO produced by V-PROLI/NO treatment. Cyp1a1 expression was directly related to the level of NO production and to reduced arsenic cytotoxicity. V-PROLI/NO pretreatment markedly reduced arsenic induced apoptosis and suppressed phosphorylation of JNK1/2. V-PROLI/NO pretreatment facilitated additional increases in arsenic-induced metallothionein, a metal-binding protein important in arsenic tolerance. Thus, V-PROLI/NO protects against arsenic toxicity in rat liver cells, reducing cytolethality, apoptosis and dysregulation of MAPKs, through generation of NO formed after metabolism by liver cell enzymes, possibly including Cyp1a1. CYP450 required for NO production from V-PROLI/NO treatment in the rat and human appears to differ as we have previously studied the ability of V-PROLI/NO to prevent arsenic toxicity in human liver cells where it reduced toxicity apparently through a CYP2E1-mediated metabolic mechanism. None-the-less, it appears that both rat and human liver cells act upon V-PROLI/NO via a CYP450-related mechanism to produce NO and subsequently reduce arsenic toxicity. PMID- 21621527 TI - Dietary astaxanthin inhibits colitis and colitis-associated colon carcinogenesis in mice via modulation of the inflammatory cytokines. AB - Astaxanthin (AX) is one of the marine carotenoid pigments, which possess powerful biological antioxidant, anti-inflammatory and anti-cancer properties. The purpose of this study is to investigate possible inhibitory effect of AX against inflammation-related mouse colon carcinogenesis and dextran sulfate sodium (DSS) induced colitis in male ICR mice. We conducted two different experiments. In the first experiment, we evaluated the effects of AX at three dose levels, 50, 100 and 200 ppm in diet, on colitis-associated colon carcinogenesis induced by azoxymethane (AOM)/DSS in mice. In the second, the effects of the AX (100 and 200 ppm) in diet on DSS-induced colitis were determined. We found that dietary AX significantly inhibited the occurrence of colonic mucosal ulcers, dysplastic crypts, and colonic adenocarcinoma at week 20. AX-feeding suppressed expression of inflammatory cytokines, including nuclear factor (NF)-kappaB, tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta, inhibited proliferation, and induced apoptosis in the colonic adenocarcinomas. Feeding with 200 ppm AX, but not 100 ppm, significantly inhibited the development of DSS-induced colitis. AX feeding (200 ppm in diet) also lowered the protein expression of NF-kappaB, and the mRNA expression of inflammatory cytokines, including IL-1beta, IL-6, and cyclooxygenase (COX)-2. Our results suggest that the dietary AX suppresses the colitis and colitis-related colon carcinogenesis in mice, partly through inhibition of the expression of inflammatory cytokine and proliferation. Our findings suggest that AX is one of the candidates for prevention of colitis and inflammation-associated colon carcinogenesis in humans. PMID- 21621528 TI - Combining body mass index and serum potassium to urine potassium clearance ratio is an alternative method to predict primary aldosteronism. AB - BACKGROUND: Though aldosterone-renin ratio (ARR) is the current routine screening method for suspicious primary aldosteronism, we hypothesized that the simple formula combining body mass index (BMI) and serum potassium to urine potassium clearance (PUKC) ratio was comparable to ARR. METHODS: Records of patients who were referred to the National Taiwan University Hospital for investigation of primary aldosteronism from January 1995 through December 2007 were retrieved. Primary aldosteronism was diagnosed based on the modified 4-corners criteria, otherwise essential hypertension was diagnosed. In both groups, the PUKC/BMI ratio was determined as well as the ARR. Bland-Altman and mountain-plot analysis were used to validate the agreement between ARR and PUKC/BMI. Receiver operating characteristic (ROC) curves were used to compare the sensitivity and specificity of PUKC/BMI and ARR. RESULTS: The records for urinary potassium were analyzed for 177 hypertensive patients (134 patients with primary aldosteronism). ROC curves showed comparable areas under the curves of both methods (95% CI: -0.029 to 0.183; p=0.186). Bland-Altman analysis further supported the agreement between ARR and PUKC/BMI ratio. CONCLUSIONS: We found that the screening power of PUKC/BMI was as good as that of conventional ARR. With the quick and extensive availability of the PUKC/BMI method and its equivalence to ARR, this screening strategy would be a good first-line tool for massive community-based primary aldosteronism surveys. PMID- 21621529 TI - Effective vitrification of human induced pluripotent stem cells using carboxylated epsilon-poly-l-lysine. AB - Derivation of human induced pluripotent stem (iPS) cells could enable their widespread application in future. Establishment of highly efficient and reliable methods for their preservation is a prerequisite for these applications. In this study, we developed a vitrification solution comprising ethylene glycol (EG) and sucrose as well as carboxylated epsilon-poly-l-lysine (PLL); this solution inhibited devitrification. Human iPS cells were vitrified in 200-MUL vitrification solutions comprised 6.5M EG, 0.75 M sucrose and 0 or 10%w/v carboxylated PLL with 65 mol% of the amino groups converted to carboxyl groups [PLL (0.65)] in a cryovial by directly immersing in liquid nitrogen. After warming, attached colony and recovery rates of human iPS cells vitrified by adding PLL (0.65) were significantly higher than those for cells without PLL (0.65) and vitrification solution (DAP213: 2M dimethyl sulfoxide, 1M acetamide and 3M propylene glycol). Furthermore, even after warming at room temperature, attached colony and recovery rates of iPS cells vitrified with PLL (0.65) were reduced to a lesser extent than those vitrified with either DAP213 or EG and sucrose without PLL (0.65). This could be attributed to inhibition of devitrification by PLL (0.65), as differential scanning calorimetry indicated less damage after vitrification with PLL (0.65). In addition, human iPS cells vitrified in the solution with PLL (0.65) had normal karyotypes and maintained undifferentiated states and pluripotency as determined by immunohistochemistry and teratoma formation. Addition of PLL (0.65) successfully vitrified human iPS cells with high efficiency. We believe that this method could aid future applications and increase utility of human iPS cells. PMID- 21621530 TI - Severe impairment of cerebellum development in mice expressing a dominant negative mutation inactivating thyroid hormone receptor alpha1 isoform. AB - Thyroid hormone deficiency is known to deeply affect cerebellum post-natal development. We present here a detailed analysis of the phenotype of a recently generated mouse model, expressing a dominant-negative TRalpha1 mutation. Although hormonal level is not affected, the cerebellum of these mice displays profound alterations in neuronal and glial differentiation, which are reminiscent of congenital hypothyroidism, indicating a predominant function of this receptor isoform in normal cerebellum development. Some of the observed effects might result from the cell autonomous action of the mutation, while others are more likely to result from a reduction in neurotrophic factor production. PMID- 21621531 TI - Pou5f1 contributes to dorsoventral patterning by positive regulation of vox and modulation of fgf8a expression. AB - Pou5f1/Oct-4 in mice is required for maintenance of embryonic pluripotent cell populations. Zebrafish pou5f1 maternal-zygotic mutant embryos (spiel ohne grenzen; MZspg) lack endoderm and have gastrulation and dorsoventral patterning defects. A contribution of Pou5f1 to the control of bmp2b, bmp4 and vox expression has been suggested, however the mechanisms remained unclear and are investigated in detail here. Low-level overexpression of a Pou5f1-VP16 activator fusion protein can rescue dorsalization in MZspg mutants, indicating that Pou5f1 acts as a transcriptional activator during dorsoventral patterning. Overexpression of larger quantities of Pou5f1-VP16 can ventralize wild-type embryos, while overexpression of a Pou5f1-En repressor fusion protein can dorsalize embryos. Lack of Pou5f1 causes a transient upregulation of fgf8a expression after mid-blastula transition, providing a mechanism for delayed activation of bmp2b in MZspg embryos. Overexpression of the Pou5f1-En repressor induces fgf8, suggesting an indirect mechanism of Pou5f1 control of fgf8a expression. Transcription of vox is strongly activated by Pou5f1-VP16 even when translation of zygotically expressed transcripts is experimentally inhibited by cycloheximide. In contrast, bmp2b and bmp4 are not activated under these conditions. We show that Pou5f1 binds to phylogenetically conserved Oct/Pou5f1 sites in the vox promoter, both in vivo (ChIP) and in vitro. Our data reveals a set of direct and indirect interactions of Pou5f1 with the BMP dorsoventral patterning network that serve to fine-tune dorsoventral patterning mechanisms and coordinate patterning with developmental timing. PMID- 21621532 TI - DMRT1 promotes oogenesis by transcriptional activation of Stra8 in the mammalian fetal ovary. AB - Dmrt1 belongs to the DM domain gene family of conserved sexual regulators. In the mouse Dmrt1 is expressed in the genital ridge (the gonadal primordium) in both sexes and then becomes testis-specific shortly after sex determination. The essential role of DMRT1 in testicular differentiation is well established, and includes transcriptional repression of the meiotic inducer Stra8. However Dmrt1 mutant females are fertile and the role of Dmrt1 in the ovary has not been studied. Here we show in the mouse that most Dmrt1 mutant germ cells in the fetal ovary have greatly reduced expression of STRA8, and fail to properly localize SYCP3 and gammaH2AX during meiotic prophase. Lack of DMRT1 in the fetal ovary results in the formation of many fewer primordial follicles in the juvenile ovary, although these are sufficient for fertility. Genome-wide chromatin immunoprecipitiation (ChIP-chip) and quantitative ChIP (qChIP) combined with mRNA expression profiling suggests that transcriptional activation of Stra8 in fetal germ cells is the main function of DMRT1 in females, and that this regulation likely is direct. Thus DMRT1 controls Stra8 sex-specifically, activating it in the fetal ovary and repressing it in the adult testis. PMID- 21621533 TI - Ephexin4 and EphA2 mediate resistance to anoikis through RhoG and phosphatidylinositol 3-kinase. AB - Disruption of cell-extracellular matrix interaction causes epithelial cells to undergo apoptosis called anoikis, and resistance to anoikis has been suggested to be a critical step for cancer cells to metastasize. EphA2 is frequently overexpressed in a variety of human cancers, and recent studies have found that overexpression of EphA2 contributes to malignant cellular behavior, including resistance to anoikis, in several different types of cancer cells. Here we show that Ephexin4, a guanine nucleotide exchange factor for the small GTPase RhoG that interacts with EphA2, plays an important role in the regulation of anoikis. Knockdown of Ephexin4 promoted anoikis in HeLa cells, and experiments using a knockdown-rescue approach showed that activation of RhoG, phosphatidylinositol 3 kinase (PI3K), and Akt was required for the Ephexin4-mediated suppression of anoikis. Indeed, Ephexin4 knockdown caused a decrease in RhoG activity and Akt phosphorylation in HeLa cells cultured in suspension. In addition, Ephexin4 was involved in the EphA2-mediated suppression of anoikis. Taken together, these results suggest that Ephexin4 mediates resistance to anoikis through activation of RhoG and PI3K downstream of EphA2. PMID- 21621534 TI - Regulation of death complexes formation in tumor necrosis factor receptor signaling. AB - TNFalpha stimulation triggers both cell death and survival programs. Since dysregulated apoptosis or cell growth can cause inflammatory diseases, cancer, or autoimmune disorders, it is important to understand the molecular mechanism of controlling cell death and survival by TNFR downstream signaling molecules. In this study, we used normal diploid cells, mouse embryonic fibroblasts (MEFs), to mimic the general TNFalpha-resistant phenomenon seen under physiological conditions.We elucidated the TNFalpha-induced death signaling complexes in TNF alpha-resistant WT MEFs and TNFalpha-sensitive MEFs that were cFLIP-, RelA-, TRAF2- or RIP1-deficient. Consistent with TNFalpha-mediated killing, we detected TNFalpha-induced high molecular weight complexes containing caspase-8 and FADD by gel filtration in the deficient MEFs, especially in those devoid of cFLIP. In addition to the presence of caspase-8-FADD in the TNFalpha-induced-death complex in the deficient MEFs, we also detected an intermediate protein complex containing RIP1, TRAF2 and caspase-8.Moreover, we demonstrated a correlation between TNFalpha-sensitivity and death-inducing complex ability in two transformed cell lines, E1A- and Ras- transformed MEFs and PDGF-B-transformed NIH 3T3 cells with PDGF-B signaling inhibited by the tyrosine kinase inhibitor STI571. Taken together, our results suggest the involvement of cFLIP-, RelA-, RIP1-, or TRAF2-related mechanisms for preventing FADD-caspase-8 interaction in wild-type MEFs. PMID- 21621535 TI - No association between the T280M polymorphism of the CX3CR1 gene and exudative AMD. AB - Major genetic factors for age-related macular degeneration (AMD) have recently been identified as susceptibility risk factors. The CX3CR1 gene has been shown to be associated with AMD in some studies. Our purpose was to analyze the role of the T280M polymorphism of the CX3CR1 gene in a large French population, in a case control study. 1093 patients with exudative AMD and 396 controls have been recruited and genotyped for the Y402H of CFH, rs10490924 of ARMS2 and T280M of the CX3CR1 gene. The distribution of the Y402H of CFH and of the rs10490924 of ARMS2 was significantly different between cases and controls (p < 0.0001). The distribution of the T280M genotypes was not significantly different in the AMD patients compared to controls (p = 0.99). The Odds Ratio compared to TT individuals was 1.0 (95% CI 0.8-1.3) for TM individuals and 1.0 (95% CI 0.5-2.1) for MM individuals. The M allele frequency was 0.157 in cases and 0.154 in controls (p = 0.87). Our study exclude an association between the T280M of the CX3CR1 gene and exudative AMD in a French population. PMID- 21621536 TI - Measurement of episcleral venous pressure. AB - Episcleral venous pressure (EVP) is an important determinant of intraocular pressure (IOP) and can be measured by using various techniques. It has been measured non-invasively by estimating the pressure required to compress an episcleral vein to a predetermined endpoint. However, the lack of objective endpoints makes EVP measurement in humans uncertain, and a wide range of mean EVP has been reported in the literature. We review the evidence for physiologic regulation of EVP and its role in glaucoma therapy, techniques that have been used to measure EVP and the need for objective measurements, and reported values for EVP. We also review recent progress toward developing an objective technique for EVP measurement. PMID- 21621537 TI - Identification of a novel DNA element that promotes cell-to-cell transformation in Escherichia coli. AB - Recently, we discovered a novel phenomenon, "cell-to-cell transformation" by which non-conjugative plasmids are transmitted horizontally in co-cultures of Escherichia coli F(-) strains. In this study, we aimed to identify the DNA element responsible for the high cell-to-cell transformability of pHSG299. By transplanting pHSG299 DNA fragments into pHSG399, a plasmid showing low transformability, we discovered that a specific 88 bp fragment of pHSG299 significantly promoted pHSG399 transformability. Although several short motif like repetitive sequences (6-10 bp) were present in the 88 bp sequence, no known DNA motifs were recognized, suggesting that this 88 bp sequence (cell-to-cell transformation promoting sequence, CTPS; Accession number: AB634455) is a novel DNA element. PMID- 21621538 TI - Histone tails: Directing the chromatin response to DNA damage. AB - Considerable energetic investment is devoted to altering large stretches of chromatin adjacent to DNA double strand breaks (DSBs). Immediately ensuing DSB formation, a myriad of histone modifications are elicited to create a platform for inducible and modular assembly of DNA repair protein complexes in the vicinity of the DNA lesion. This complex signaling network is critical to repair DNA damage and communicate with cellular processes that occur in cis and in trans to the genomic lesion. Failure to properly execute DNA damage inducible chromatin changes is associated with developmental abnormalities, immunodeficiency, and malignancy in humans and in genetically engineered mouse models. This review will discuss current knowledge of DNA damage responsive histone changes that occur in mammalian cells, highlighting their involvement in the maintenance of genome integrity. PMID- 21621539 TI - Endoscopic mucosal resection: not your father's polypectomy anymore. PMID- 21621541 TI - The Australian lungfish, Neoceratodus forsteri: a personal story. AB - The following is a brief description of how lungfish research at Macquarie University began, of the period in which it flourished, and, most recently, of the winding down of the University's involvement with this research. During this latter period, the Australian lungfish in the wild were threatened by the construction of a megadam in their very limited habitat. Fortunately, this was averted in December 2009, after 3 years of lobbying the Federal Government. They now await another "Aussie" to make them accessible for further research by Australian and international researchers. PMID- 21621542 TI - Mechanisms responsible for progesterone's protection against lordosis-inhibiting effects of restraint II. Role of progesterone metabolites. AB - When ovariectomized Fischer female rats are hormonally primed with 10 MUg estradiol benzoate, a 5 min restraint experience rapidly inhibits lordosis behavior. Addition of progesterone to the hormonal priming prevents this restraint-induced inhibition. In prior work, we reported evidence that progesterone receptors (PR) may contribute to this protective effect of progesterone. In the current manuscript, we provide evidence that progesterone metabolites may also contribute to progesterone's ability to reduce the effects of restraint. Ovariectomized female rats were hormonally primed with 10 MUg estradiol benzoate followed 2 days later with 4.0 mg/kg of the progesterone metabolite, allopregnanolone. Allopregnanolone, administered either 4 h or 2 h before the restraint experience, was as effective as progesterone in reducing the lordosis-inhibitory effects of restraint. In the second experiment, progesterone metabolism was blocked with 50 mg/kg of the 5alpha-reductase inhibitor, finasteride. Surprisingly, finasteride did not prevent progesterone from reducing the effects of restraint. In a third experiment, we tested the possibility that allopregnanolone acted through metabolism to dihydroprogesterone. Rats were treated with allopregnanolone or with allopregnanolone plus the 3alpha hydroxysteroid dehydrogenase inhibitor, indomethacin. Indomethacin did not prevent allopregnanolone from reducing the effects of restraint. Mechanisms are discussed whereby cross-talk between PR-mediated and metabolite-mediated events may converge in producing progesterone's attenuation of the effect of restraint. PMID- 21621543 TI - Nosema ceranae in drone honey bees (Apis mellifera). AB - Nosema ceranae is a microsporidian intracellular parasite of honey bees, Apis mellifera. Previously Nosema apis was thought to be the only cause of nosemosis, but it has recently been proposed that N. ceranae is displacing N. apis. The rapid spread of N. ceranae could be due to additional transmission mechanisms, as well as higher infectivity. We analyzed drones for N. ceranae infections using duplex qPCR with species specific primers and probes. We found that both immature and mature drones are infected with N. ceranae at low levels. This is the first report detecting N. ceranae in immature bees. Our data suggest that because drones are known to drift from their parent hives to other hives, they could provide a means for disease spread within and between apiaries. PMID- 21621540 TI - Twin study indicates loss of interaction between microbiota and mucosa of patients with ulcerative colitis. AB - BACKGROUND & AIMS: Interactions between genetic and environmental factors are believed to be involved in onset and initiation of inflammatory bowel disease. We analyzed the interaction between gastrointestinal mucosal microbiota and host genes in twin pairs discordant for ulcerative colitis (UC) to study the functional interaction between microbiota and mucosal epithelium. METHODS: Biopsy were collected from sigmoid colon of UC patients and their healthy twins (discordant twin pairs) and from twins without UC. Microbiota profiles were determined from analysis of 16S ribosomal DNA libraries; messenger RNA profiles were determined by microarray analysis. RESULTS: Patients with UC had dysbiotic microbiota, characterized by less bacterial diversity and more Actinobacteria and Proteobacteria than that of their healthy siblings; healthy siblings from discordant twins had more bacteria from the Lachnospiraceae and Ruminococcaceae families than twins who were both healthy. In twins who were both healthy, 34 mucosal transcripts correlated with bacterial genera, whereas only 25 and 11 correlated with bacteria genera in healthy individuals and their twins with UC, respectively. Transcripts related to oxidative and immune responses were differentially expressed between patients with UC and their healthy twins. CONCLUSIONS: The transcriptional profile of the mucosa appears to interact with the colonic microbiota; this interaction appears to be lost in colon of patients with UC. Bacterial functions, such as butyrate production, might affect mucosal gene expression. Patients with UC had different gene expression profiles and lower levels of biodiversity than their healthy twins, as well as unusual aerobic bacteria. Patients with UC had lower percentages of potentially protective bacterial species than their healthy twins. PMID- 21621544 TI - A new apo-caspase-6 crystal form reveals the active conformation of the apoenzyme. AB - Caspase-6 has been identified as a key component in the pathway of neurodegenerative diseases such as Alzheimer's disease and Huntington's disease. It has been the focus of drug development for some time, but only recently have structural data become available. The first study identified a novel noncanonical conformation of apo-caspase-6 contrasting with the typical caspase conformation. Then, the structures of both caspase-6 zymogen and the Ac-VEID-CHO peptide inhibitor complex described caspase-6 in the canonical conformation, raising the question of why the intermediate between these two structures (mature apo-caspase 6) would adopt the noncanonical conformation. In this study, we present a new crystal form of the apoenzyme in the canonical conformation by identifying the previous apostructure as a pH-inactivated form of caspase-6. Our new apostructure is further compared to the Ac-VEID-CHO caspase-6 inhibitor complex. The structural comparison allows us to visualize the organization of loops L2, L3, and L4 upon ligand binding and how the catalytic groove forms to accommodate the inhibitor. PMID- 21621545 TI - Structural basis for Abeta1-42 toxicity inhibition by Abeta C-terminal fragments: discrete molecular dynamics study. AB - Amyloid beta-protein (Abeta) is central to the pathology of Alzheimer's disease. Of the two predominant Abeta alloforms, Abeta(1-40) and Abeta(1-42), the latter forms more toxic oligomers. C-terminal fragments (CTFs) of Abeta were recently shown to inhibit Abeta(1-42) toxicity in vitro. Here, we studied Abeta(1-42) assembly in the presence of three effective CTF inhibitors and an ineffective fragment, Abeta(21-30). Using a discrete molecular dynamics approach that recently was shown to capture key differences between Abeta(1-40) and Abeta(1-42) oligomerization, we compared Abeta(1-42) oligomer formation in the absence and presence of CTFs or Abeta(21-30) and identified structural elements of Abeta(1 42) that correlated with Abeta(1-42) toxicity. CTFs co-assembled with Abeta(1-42) into large heterooligomers containing multiple Abeta(1-42) and inhibitor fragments. In contrast, Abeta(21-30) co-assembled with Abeta(1-42) into heterooligomers containing mostly a single Abeta(1-42) and multiple Abeta(21-30) fragments. The CTFs, but not Abeta(21-30), decreased the beta-strand propensity of Abeta(1-42) in a concentration-dependent manner. CTFs and Abeta(21-30) had a high binding propensity to the hydrophobic regions of Abeta(1-42), but only CTFs were found to bind the Abeta(1-42) region A2-F4. Consequently, only CTFs but not Abeta(21-30) reduced the solvent accessibility of Abeta(1-42) in region D1-R5. The reduced solvent accessibility of Abeta(1-42) in the presence of CTFs was comparable to the solvent accessibility of Abeta(1-40) oligomers formed in the absence of Abeta fragments. These findings suggest that region D1-R5, which was more exposed to the solvent in Abeta(1-42) than in Abeta(1-40) oligomers, is involved in mediating Abeta(1-42) oligomer neurotoxicity. PMID- 21621546 TI - Targeting capacity and conservation of PreP homologues localization in mitochondria of different species. AB - Mitochondrial presequences and other unstructured peptides are degraded inside mitochondria by presequence proteases (PrePs) identified in Arabidopsis thaliana (AtPreP), humans (hPreP), and yeast (Cym1/Mop112). The presequences of A. thaliana and human PreP are predicted to consist of 85 and 29 amino acids, respectively, whereas the Saccharomyces cerevisiae Cym1/Mop112 presequence contains only 7 residues. These differences may explain the reported targeting of homologous proteins to different mitochondrial subcompartments. Here we have investigated the targeting capacity of the PreP homologues' presequences. We have produced fusion constructs containing N-terminal portions of AtPreP(1-125), hPreP(1-69), and Cym1(1-40) coupled to green fluorescent protein (GFP) and studied their import into isolated plant, mammalian, and yeast mitochondria, followed by mitochondrial subfractionation. Whereas the AtPreP presequence has the capacity to target GFP into the mitochondrial matrix of all three species, the hPreP presequence only targets GFP to the matrix of mammalian and yeast mitochondria. The Cym1/Mop112 presequence has an overall much weaker targeting capacity and only ensures mitochondrial sorting in its host species yeast. Revisiting the submitochondrial localization of Cym1 revealed that endogenous Cym1/Mop112 is localized to the matrix space, as has been previously reported for the plant and human homologues. Moreover, complementation studies in yeast show that native AtPreP restores the growth phenotype of yeast cells lacking Cym1, demonstrating functional conservation. PMID- 21621547 TI - Epididymal lithiasis in roosters: in the middle of the way there was a stone. AB - The epididymal region plays an important role in the reproduction of roosters, as it is the site of functions important in the maintenance of fertility, including fluid and calcium reabsorption and sperm surface modifications. About 10 years ago, a reproductive dysfunction characterized by the formation of luminal calcium stones in the epididymal region of roosters was described. This anomaly, known as epididymal lithiasis, is associated with a significant decrease in the fertility of affected roosters. This reproductive anomaly has been observed in multiple countries and is thought to negatively impact the poultry industry; however, the cause of epididymal lithiasis has not been fully determined. Several hypotheses have been proposed to explain the origin of epididymal lithiasis, including the presence of an infectious agent within the epididymal region, an autoimmune response, increased dietary calcium and vitamin D3 intake and the presence of genetic susceptibility factors; however, none of these has been proven to be the primary cause of the calcium stone formation. Nonetheless, considerable evidence suggests that regardless of the primary cause of epididymal lithiasis, this anomaly could result from a hormonal imbalance or a local impairment of calcium homeostasis in the epididymal region. The objectives of this mini-review are to 1) summarize the reproductive alterations observed in animals affected by epididymal lithiasis, 2) discuss the hypotheses proposed to explain the cause of luminal stone formation and 3) provide perspectives for future studies of this reproductive disorder. PMID- 21621548 TI - Dopamine system dysregulation by the hippocampus: implications for the pathophysiology and treatment of schizophrenia. AB - For decades, the predominant hypothesis of schizophrenia centered on dysfunctions of the dopamine system. However, recent evidence now suggests that the dopamine system may be "normal" in its configuration, but instead is regulated abnormally by modulatory processes. Convergent studies in animals and in humans have now focused on the hippocampus as a central component in the generation of psychosis and possibly other symptom states in schizophrenia. Thus, activity in the ventral hippocampus has been shown to regulate dopamine neuron responsivity by controlling the number of dopamine neurons that can be phasically activated by stimuli. In this way, this structure determines the gain of the dopamine signal in response to stimuli. However, in schizophrenia, the hippocampus appears to be hyper-active, possibly due to attenuation of function of inhibitory interneurons. As a result, the dopamine system is driven into an overly responsive state. Current medications have focused on blockade of overstimulated dopamine receptors; however, this now appears to be several synapses downstream from the pathological antecedent. Therapeutic approaches that focus on normalizing hippocampal function may prove to be more effective treatment avenues for the schizophrenia patient. PMID- 21621549 TI - The impact of overnight consolidation upon memory for emotional and neutral encoding contexts. AB - Sleep plays a role in the consolidation of declarative memories. Although this influence has attracted much attention at the level of behavioural performance, few reports have searched for neural correlates. Here, we studied the impact of sleep upon memory for the context in which stimuli were learned at both behavioural and neural levels. Participants retrieved the association between a presented foreground object and its encoding context following a 12-h retention interval including either wake only or wake plus a night of sleep. Since sleep has been shown to selectively enhance some forms of emotional memory, we examined both neutral and emotionally valenced contexts. Behaviourally, less forgetting was observed across retention intervals containing sleep than retention intervals containing only wakefulness, and this benefit was accompanied by stronger responses in hippocampus and superior parietal cortex. This sleep-related reduction in forgetting did not differ between neutral and negative contexts, but there was a clear interaction between sleep and context valence at the functional level, with left amygdala, right parahippocampus, and other components of the episodic memory system all responding more strongly during correct memory for emotional contexts post-sleep. Connectivity between right parahippocampus and bilateral amygdala/periamygdala was also enhanced during correct post-sleep attribution of emotional contexts. Because there was no interaction between sleep and valence in terms of context memory performance these functional results may be associated with memory for details about the emotional encoding context rather than for the link between that context and the foreground object. Overall, our data show that while context memory decays less across sleep than across an equivalent period of wake, the sleep-related protection of such associations is not influenced by context emotionality in the same way as direct recollection of emotional information. PMID- 21621550 TI - A helminth cestode parasite express an estrogen-binding protein resembling a classic nuclear estrogen receptor. AB - The role of an estrogen-binding protein similar to a known mammalian estrogen receptor (ER) is described in the estradiol-dependent reproduction of the helminth parasite Taenia crassiceps. Previous results have shown that 17-beta estradiol induces a concentration-dependent increase in bud number of in vitro cultured cysticerci. This effect is inhibited when parasites are also incubated in the presence of an ER binding-inhibitor (tamoxifen). RT-PCR assays using specific oligonucleotides of the most conserved ER sequences, showed expression by the parasite of a mRNA band of molecular weight and sequence corresponding to an ER. Western blot assays revealed reactivity with a 66 kDa protein corresponding to the parasite ER protein. Tamoxifen treatment strongly reduced the production of the T. crassiceps ER-like protein. Antibody specificity was demonstrated by immunoprecipitating the total parasite protein extract with anti ER-antibodies. Cross-contamination by host cells was discarded by flow cytometry analysis. ER was specifically detected on cells expressing paramyosin, a specific helminth cell marker. Parasite cells expressing the ER-like protein were located by confocal microscopy in the subtegumental tissue exclusively. Analysis of the ER-like protein by bidimensional electrophoresis and immunoblot identified a specific protein of molecular weight and isoelectric point similar to a vertebrates ER. Sequencing of the spot produced a small fragment of protein similar to the mammalian nuclear ER. Together these results show that T. crassiceps expresses an ER-like protein which activates the budding of T. crassiceps cysticerci in vitro. To the best of our knowledge, this is the first report of an ER-like protein in parasites. This finding may have strong implications in the fields of host-parasite co-evolution as well as in sex associated susceptibility to this infection, and could be an important target for the design of new drugs. PMID- 21621551 TI - Fipronil is a powerful uncoupler of oxidative phosphorylation that triggers apoptosis in human neuronal cell line SHSY5Y. AB - Fipronil is a phenylpyrazole insecticide known to elicit neurotoxicity via an interaction with ionotropic receptors, namely GABA and glutamate receptors. Recently, we showed that fipronil and other phenylpyrazole compounds trigger cell death in Caco-2 cells. In this study, we investigated the mode of action and the type of cell death induced by fipronil in SH-SY5Y human neuroblastoma cells. Flow cytometric and western blot analyses demonstrated that fipronil induces cellular events belonging to the apoptosis process, such as mitochondrial potential collapse, cytochrome c release, caspase-3 activation, nuclear condensation and phosphatidylserine externalization. In addition, fipronil induces a rapid ATP depletion with concomitant activation of anaerobic glycolysis. This cellular response is characteristic of mitochondrial injury associated with a defect of the respiration process. Therefore, we also investigated the effect of fipronil on the oxygen consumption in isolated mitochondria. Interestingly, we show for the first time that fipronil is a strong uncoupler of oxidative phosphorylation at relative low concentrations. Thus in this study, we report a new mode of action by which the insecticide fipronil could triggers apoptosis. PMID- 21621552 TI - Induction of regulatory T cells by green tea polyphenol EGCG. AB - Regulatory T cells (Treg) are critical in maintaining immune tolerance and suppressing autoimmunity. The transcription factor Foxp3 serves as a master switch that controls the development and function of Treg. Foxp3 expression is epigenetically regulated by DNA methylation, and DNA methyltransferase (DNMT) inhibitors can induce Foxp3 expression in naive CD4(+) T cells. We showed that EGCG, a major green tea polyphenol, could act as a dietary DNMT inhibitor, and induced Foxp3 and IL-10 expression in CD4(+) Jurkat T cells at physiologically relevant concentrations in vitro. We further showed that mice treated with EGCG in vivo had significantly increased Treg frequencies and numbers in spleen and lymph nodes and had inhibited T cell response. Induction of Foxp3 expression correlated with a concomitant reduction in DNMT expression and a decrease in global DNA methylation. Our data suggested that EGCG can induce Foxp3 expression and increase Treg frequency via a novel epigenetic mechanism. While the DNMT inhibitory effects of EGCG was not as potent as pharmacologic agents such as 5 aza-2'-deoxycytidine, the ability of dietary agents to target similar mechanisms offers opportunities for potentially sustained and longer-term exposures with lower toxicity. Our work provides the foundation for future studies to further examine and evaluate dietary strategies to modulate immune function. PMID- 21621553 TI - Interleukin-2 rescues helpless effector CD8+ T cells by diminishing the susceptibility to TRAIL mediated death. AB - CD8(+) T cells primed in the absence of CD4(+) T cell help are programmed to produce TRAIL, which results in Death receptor (DR5) mediated apoptosis upon restimulation. Here, we studied whether these 'helpless' effector CD8(+) T cells are consigned to an apoptotic fate or whether their helpless program can be altered by inflammatory or growth cytokines. We found that helpless CD8(+) T cells regained their full proliferative and functional capacity only when IL-2 was added to cell cultures, while IL-7 and IL-15, two common gamma chain cytokines associated with CD8(+) T cell homeostasis and memory, could only partly restore secondary expansion in helpless CD8(+) T cells. Recovery of functional CD8(+) T cell immunity by IL-2 was concomitant with induction of IL2Ralpha (CD25) expression, downregulation of TRAIL, and the upregulation of anti-apoptotic molecules Bcl-2 and FLIP. The addition of IL-2 to helpless CD8(+) T cells also interfered with DR5-mediated apoptosis induction, indicating that IL-2 affects several components of the TRAIL-DR5 pathway. Collectively, these data demonstrate that the helpless phenotype is not fixed, and that IL-2R signaling at the time of reactivation can play an important role in restoring CD8(+) T cell function. PMID- 21621554 TI - Visual detection of human enterovirus 71 subgenotype C4 and Coxsackievirus A16 by reverse transcription loop-mediated isothermal amplification with the hydroxynaphthol blue dye. AB - A sensitive reverse transcription loop-mediated isothermal amplification (RT LAMP) assay was developed for rapid visual detection of human enterovirus 71 subgenotype C4 (EV71-C4) and Coxsackievirus A16 (CVA16) infection, respectively. The reaction was performed in one step in a single tube at 65 degrees C for 60 min with the addition of the hydroxynaphthol blue (HNB) dye prior to amplification. The detection limits of the RT-LAMP assay were 0.33 and 1.58 of a 50% tissue culture infective dose (TCID(50)) per reaction based on 10-fold dilutions of a titrated EV71 or CVA16 strain, respectively. No cross-reaction was observed with Coxsackievirus A (CVA) viruses (CVA2, 4, 5, 7, 9, 10, 14, and 24), Coxsackievirus B (CVB) viruses (CVB1,2,3,4, and 5) or ECHO viruses (ECHO3, 6, 11, and 19). The assay was further evaluated with 47 clinical stool specimens diagnosed previously with EV71, CVA16 or other human enterovirus infections. Virus isolates from stool samples were confirmed by virus neutralization testing and sequencing. RT-LAMP with HNB dye was demonstrated to be a sensitive and cost effective assay for rapid visual detection of human EV71-C4 and CVA16. PMID- 21621555 TI - Single dilution Avidity-Blocking ELISA as an alternative to the Bovine Viral Diarrhea Virus neutralization test. AB - This study describes the development and validation of a blocking ELISA that measures avidity of BVDV-specific immunoglobulins (Igs) as an alternative to the classic virus neutralization test. The assay comprises a recombinant soluble E2 glycoprotein as target antigen, a neutralizing serum as detector antibody and a washing-step with a chaotropic agent to determine BVDV-specific Igs avidity. Avidity-Blocking ELISA was validated with 100 negative and 87 positive BVDV neutralization serum samples from either infected or vaccinated bovines (inactivated commercial vaccines). Specificity and sensitivity of the Avidity Blocking ELISA were 100% and 98.8%, respectively. The assay was standardized to use a single dilution, so that 90 samples can be tested per plate. Results expressed as Avidity Index (AI) correlated with BVDV neutralizing titers (r=0.94). Unlike the virus neutralization test, the Avidity-Blocking ELISA could discriminate between infected and vaccinated animals (DIVA), suggesting that avidity measurement can be a valuable tool to achieve DIVA compliances. The data show that the avidity of anti BVDV antibodies is related to their capacity to block viral infection in vitro. PMID- 21621556 TI - Aphids preserved in propylene glycol can be used for reverse transcription polymerase chain reaction detection of Potato virus Y. AB - The effectiveness of propylene glycol on the retention of RNA target of Potato virus Y (PVY), an aphid stylet-borne virus, in Myzus persicae was investigated in comparison to ethanol and liquid nitrogen/-80 degrees C. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the PVY targets from the propylene glycol/ethanol/liquid nitrogen preserved single aphids after a 5min acquisition period from infected potato plants. In the liquid nitrogen/-80 degrees C and 70% ethanol treatments, 55.6% and 38.8% aphids tested PVY-positive, respectively. In the 0-75% propylene glycol treatments, 12.2-44.7% aphids tested PVY-positive. The lowest detection rate was in the 0% (positive rate, 15.2%) and the 10% propylene glycol (positive rate, 12.2%). As the propylene glycol concentration increased to 25%, 29.8% aphids tested positive. A high PVY-positive rate was also found in 35-75% propylene glycol treatments at 44.7% (35% propylene glycol), 36.7% (50% propylene glycol) and 34.8% (75% propylene glycol), which is comparable to the rate shown in 70% ethanol. No significant difference in the positive detection rate was observed in aphids preserved in 50% propylene glycol at room temperature for 2, 4 and 10 days. These results demonstrate that propylene glycol at 25-75% can retain PVY targets effectively in aphids for an extended time period, and thus can be used in aphid traps to preserve viruliferous aphids for later RT-PCR detection of PVY. PMID- 21621557 TI - Cognitive side effects of cancer therapy demonstrate a functional role for adult neurogenesis. AB - Cancer therapies frequently result in a spectrum of neurocognitive deficits that include impaired learning, memory, attention and speed of information processing. Damage to dynamic neural progenitor cell populations in the brain are emerging as important etiologic factors. Radiation and chemotherapy-induced damage to neural progenitor populations responsible for adult hippocampal neurogenesis and for maintenance of subcortical white matter integrity are now believed to play major roles in the neurocognitive impairment many cancer survivors experience. PMID- 21621558 TI - Novel claustrum activation observed during a visuomotor adaptation task using a viewing window paradigm. AB - Previous literature has reported a wide range of anatomical correlates when participants are required to perform a visuomotor adaptation task. However, traditional adaptation tasks suffer a number of inherent limitations that may, in part, give rise to this variability. For instance, the sparse visual environment does not map well onto conditions in which a visuomotor transformation would normally be required in everyday life. To further clarify these neural underpinnings, functional magnetic resonance imaging (fMRI) was performed on 17 (6M, age range 20-45 years old; mean age=26) naive participants performing a viewing window task in which a visuomotor transformation was created by varying the relationship between the participant's movement and the resultant movement of the viewing window. The viewing window task more naturally replicates scenarios in which haptic and visual information would be combined to achieve a higher level goal. Even though activity related to visuomotor adaptation was found within previously reported regions of the parietal lobes, frontal lobes, and occipital lobes, novel activation patterns were observed within the claustrum - a region well-established as multi-modal convergence zone. These results confirm the diversity in the number and location of neurological systems recruited to perform a required visuomotor adaptation, and provide the first evidence of participation of the claustrum to overcome a visuomotor transformation. PMID- 21621559 TI - Is there a role for young hippocampal neurons in adaptation to stress? AB - The hippocampus has been implicated in many cognitive and emotional behaviors and in the physiology of the stress response. Within the hippocampus, the dentate gyrus has been implicated in the detection of novelty. The dentate is also a major target for stress hormones and modulates the hypothalamic-pituitary-adrenal (HPA) axis response to stress. Whether these functions of the dentate integrate or segregate remains unknown, as most investigations of its role in stress and learning are separate. Since the exciting discovery of adult neurogenesis in the dentate gyrus, adult-born neurons have been implicated in both novelty detection and the stress response. In this perspective we will discuss the literature that implicates the hippocampus, and potentially, adult-born neurons in these two functions. We will attempt to reconcile the seemingly contradictory behavioral results for the function of adult-born neurons. Finally, we will speculate that a key function of adult-born neurons within hippocampal function may be to modulate the stress response and perhaps assign stress salience to the sensory context. PMID- 21621560 TI - Exercise prevents sleep deprivation-associated anxiety-like behavior in rats: potential role of oxidative stress mechanisms. AB - Our previous work suggests that pharmacological induction of oxidative stress causes anxiety-like behavior in rats. Interestingly, sleep deprivation is reported to cause oxidative damage in the brain and is also reported to be anxiogenic. Minimal mechanistic insights are available. In this study, using a behavioral and biochemical approach, we investigated involvement of oxidative stress mechanisms in sleep deprivation-induced anxiety-like behavior of rats and the protective role of treadmill exercise in this process. We report that acute sleep deprivation (SD) increases oxidative stress in the cortex, hippocampus and amygdala while prior treadmill exercise prevents this increase. Serum corticosterones also increase with SD but its levels are normalized in exercised sleep-deprived rats. Also, anxiety-like behavior of rats significantly increases with SD while prior treadmill exercise prevents this increase. Protein expression of two enzymes involved in antioxidant defense, glyoxalase (GLO)-1 and glutathione reductase (GSR)-1 increased after 24h SD in the hippocampus, cortex and amygdala while their levels were normalized in exercised sleep-deprived rats. It is plausible that oxidative stress via regulation of GLO1 and GSR1 is involved in sleep deprivation-induced anxiety-like behavior of rats. PMID- 21621561 TI - The N-terminal alanine-extended GLP-1/IgG-Fc fusion protein confers resistance to DPP-IV and reduces serum glucose level in db/db mice. AB - The aim of this study was to develop novel long-acting glucagon-like peptide 1 (GLP-1) analogs resistant to dipeptidyl peptidase-IV (DPP-IV). We constructed three fusion proteins comprising GLP-1 and the human immunoglobulin gamma heavy chain (IgG-Fc); wild-type GLP-1 and IgG-Fc (GLP-1/IgG-Fc) and two N-terminal extended fusion proteins in which an additional Ala (A) or Gly (G) was located on the N-terminus of GLP-1 (A-GLP-1/IgG-Fc or G-GLP-1/IgG-Fc). The fusion proteins expressed in CHO-K1 cells were secreted into medium and purified by Protein A affinity chromatography. Here, we show that the Ala or Gly-extended GLP-1/IgG-Fc fusion protein is resistant to DPP-IV and has increased half-life in vivo. To our surprise, the A-GLP-1/IgG-Fc fusion protein was more effective than wildtype GLP 1/IgG-Fc fusion protein in reducing blood glucose levels in db/db mice. Our findings suggest that the A-GLP-1/IgG-Fc fusion protein could be a potential long acting GLP-1 receptor agonist for the treatment of insulin-resistant type 2 diabetes. PMID- 21621562 TI - Long and short distance movements of beta(2)-adrenoceptor in cell membrane assessed by photoconvertible fluorescent protein dendra2-beta(2)-adrenoceptor fusion. AB - Local movements of receptors in the plasma membrane have been extensively studied, as it is generally believed that the dynamics of membrane distribution of receptors regulate their functions. However, the properties of large-scale (>5MUm) receptor movements in the membrane are relatively obscure. In the present study, we addressed the question as to whether the large-scale movement of receptor in the plasma membrane at the whole cell level can be explained quantitatively by its local diffusive properties. We used HEK 293 cells transfected with human beta2-adrenoceptor fused to photoconvertible fluorescent protein dendra2 as a model system; and found that 1) functional integrity of the dendra2-tagged receptor remains apparently intact; 2) in a mesoscopic scale (~4MUm), ~90% of the receptors are mobile on average, and receptor influx to, and out-flux from a membrane area can be symmetrically explained by a diffusion-like process with an effective diffusion coefficient of ~0.1MUm(2)/s; 3) these mobility parameters are not affected by the activity state of the receptor (assessed by using constitutively active receptor mutants); 4) in the macroscopic scale (4-40MUm), although a slowly diffusing fraction of receptors (with D<0.01MUm(2)/s) is identifiable in some cases, the movement of the predominant fraction is perfectly explained by the same effective diffusion process observed in the mesoscopic scale, suggesting that the large scale structure of the cell membrane as felt by the receptor is apparently homogeneous in terms of its mesoscopic properties. We also showed that intracellular compartments and plasma membrane are kinetically connected even at steady-state. PMID- 21621563 TI - Bimodal regulation of FoxO3 by AKT and 14-3-3. AB - FoxO3 is a member of FoxO family transcription factors that mediate cellular functions downstream of AKT. FoxO3 phosphorylation by AKT generates binding sites for 14-3-3, which in-turn regulates FoxO3 transcriptional activity and localization. We examine here the functional significance of AKT-FoxO3 interaction and further detail the mechanistic aspects of FoxO3 regulation by AKT and 14-3-3. Our data show that AKT overexpression increases the steady-state levels of FoxO3 protein in a manner dependent on AKT activity and its ability to bind FoxO3. Characterization of the AKT-FoxO3 interaction shows that the three AKT phosphorylation-site-recognition motifs (RxRxxS/T) present on FoxO3, which are required for FoxO3 phosphorylation, are dispensable for AKT binding, suggesting that AKT has a docking point on FoxO3 distinct from the phosphorylation-recognition motifs. Development of a FoxO3 mutant deficient in 14 3-3 binding (P34A), which can be phosphorylated by AKT, established that 14-3-3 binding and not AKT phosphorylation per se controls FoxO3 transcriptional activity. Intriguingly, 14-3-3 binding was found to stabilize FoxO3 by inhibiting its dephosphorylation and degradation rates. Collectively, our data support a model where both AKT and 14-3-3 positively regulate FoxO3 in addition to their established negative roles and that 14-3-3 availability could dictate the fate of phosphorylated FoxO3 toward degradation or recycling. PMID- 21621564 TI - Investigation of the molecular ion structure for aldononitrile acetate derivatized muramic acid. AB - The muramic acid assay is a powerful tool for detecting both intact bacteria and bacterial debris. Past use of aldononitrile acetate derivatization for determining muramic acid in complex samples by gas chromatography/mass spectrometry met detection needs in many instances; however, questions have been raised regarding the interpretation of the derivative structure and its electron ionization fragments. In this study, we applied different methods and proved that the aldononitrile acetate derivatized muramic acid yields a molecular weight of 398, associated with a lactam structure. We also presented evidence that the structure of aldononitrile acetate derivatized muramic acid is acetylated at four positions, 3 O-acetylations and 1N-acetylation. In practical manner, this communication provides a comprehensive reference to researchers using delta(13)C value or ion fragments of the muramic acid marker in biogeochemical studies. PMID- 21621565 TI - CARD-FISH and confocal laser scanner microscopy to assess successional changes of the bacterial community in freshwater biofilms. AB - Bacterial community composition was assessed during riverine biofilm development by the Catalyzed Reporter Deposition Fluorescence In Situ Hybridization (CARD FISH) in combination with Confocal Laser Scanning Microscopy. Using artificial substrates, it was possible to follow the dynamics of specific bacterial clusters, while maintaining the unaltered structure and architecture of the biofilm. PMID- 21621566 TI - Correlation between stimulation strength and onset time of signal traveling within the neocortical neural circuits under caffeine application. AB - In general, strength of input to neocortical neural circuits affects the amplitude of postsynaptic potentials (PSPs), thereby modulating the way signals are transmitted within the circuits. Caffeine is one of the pharmacological agents able to modulate synaptic activities. The present study investigated how strength of input affects signal propagation in neocortical circuits under the application of caffeine. Spatio-temporal neural activities were observed from visual cortical slices of rats using optical recording methods with voltage sensitive dye. Electrical stimulations were applied to white matter in the primary visual cortex with bath-application of caffeine. When the strength of stimulation was 0.3mA, signals propagated from the site of stimulation in the primary visual cortex toward the secondary visual cortex along vertical and horizontal pathways. When stimulation strength was reduced from 0.3mA to 0.07mA, start of signal propagation was delayed about 25ms without affecting field PSP amplitude or the manner of signal propagation. Conversely, co-application of caffeine and d-2-amino-5-phosphonovaleric acid (d-AP5) did not induce delays in signal start. These findings suggest that conversion of neural code from amplitude code to temporal code is inducible at the level of neocortical circuits in an N-methyl-d-aspartate (NMDA) receptor activity-dependent manner. PMID- 21621567 TI - Modelling virus- and host-limitation in vectored plant disease epidemics. AB - Models of plant virus epidemics have received less attention than those caused by fungal pathogens. Intuitively, the fact that virus diseases are systemic means that the individual diseased plant can be considered as the population unit which simplifies modelling. However, the fact that a vector is required in the vast majority of cases for virus transmission, means that explicit consideration must be taken of the vector, or, the involvement of the vector in the transmission process must be considered implicitly. In the latter case it is also important that within-plant processes, such as virus multiplication and systemic movement, are taken into account. In this paper we propose an approach based on the linking of transmission at the population level with virus multiplication within plants. The resulting models are parameter-sparse and hence simplistic. However, the range of model outcomes is representative of field observations relating to the apparent limitation of epidemic development in populations of healthy susceptible plants. We propose that epidemic development can be constrained by virus limitation in the early stages of an epidemic when the availability of healthy susceptible hosts is not limiting. There is an inverse relationship between levels of transmission in the population and the mean virus titre/infected plant. In the case of competition between viruses, both virus and host limitation are likely to be important in determining whether one virus can displace another or whether both viruses can co-exist in a plant population. Lotka-Volterra type equations are derived to describe density-dependent competition between two viruses multiplying within plants, embedded within a population level epidemiological model. Explicit expressions determining displacement or co existence of the viruses are obtained. Unlike the classical Lotka-Volterra competition equations, the co-existence requirement for the competition coefficients to be both less than 1 can be relaxed. PMID- 21621568 TI - Immunity to Rice black streaked dwarf virus, a plant reovirus, can be achieved in rice plants by RNA silencing against the gene for the viroplasm component protein. AB - The nonstructural protein P9-1 of Rice black streaked dwarf virus has been confirmed to accumulate in viroplasms, the putative sites of viral replication, in infected plants and insects. We transformed rice plants by introducing an RNA interference construct against the P9-1-encoding gene. The resultant transgenic plants accumulated short interfering RNAs specific to the construct. All progenies produced by self-fertilization of these transgenic plants with induced RNA interference against the gene for P9-1 were resistant to infection by the virus. Our results demonstrated that interfering with the expression of a viroplasm component protein of plant reoviruses, which plays an important role in viral proliferation, might be a practical and effective way to control plant reovirus infection in crop plants. PMID- 21621569 TI - Improved paramagnetic liposomes for MRI visualization of pH triggered release. AB - This work aims at assessing the in vitro potential of paramagnetic pH sensitive liposomes as imaging tools for visualizing drug-delivery and release processes by Magnetic Resonance Imaging (MRI). pH sensitive liposomes (pSLs) were formulated using the fusogenic phospholipid 1-palmitoyl-2-oleoyl-sn-glycero-3 phosphoethanolamine (POPE), the membrane stabilizer D-alpha-tocopherol hemisuccinate (THS), and were loaded with several paramagnetic complexes including the clinically approved Gadoteridol (marketed as ProHanceTM). The proposed formulation allows the fast and full release of Gadoteridol at pH 5.5. The leakage of the imaging reporter from the vesicles was associated with a relaxivity enhancement that allowed its visualization by MRI. It was observed that the release mechanism implies the protonation of the THS basic sites that leads to vesicle aggregation, thus enabling the expression of the fusogenic property of POPE. Attempts for improving the MRI properties of pSLs were pursued through the encapsulation of imaging agents with higher relaxivity than Gadoteridol, but it was observed that the release kinetic can be significantly affected by the probe size. Aiming at preparing stealth pSLs, PEG chains were conjugated to the external surface of the vesicles via cleavable disulphide bridges. Such nanomedicines do not release their content at acidic pH as long as the coating polymer is not removed from the surface. The results obtained suggest that the liposomal formulation investigated in this work has the potential for visualizing drug-delivery and release processes by in vivo MRI preclinical studies. PMID- 21621570 TI - Engineering of amphiphilic block copolymers for polymeric micellar drug and gene delivery. AB - The use of nano-delivery systems formed through assembly of synthetic amphiphilic block copolymers (ABCs) in experimental medicine and pharmaceutical sciences is experiencing rapid development. This rapid development is driven by a crucial need in improving the performance of existing therapeutic agents, as well as the necessity for the development of advanced delivery systems for complex new entities such as genes, proteins and other cellular components. The flexibility in the construction of appropriate carriers for the delivery requirements of these complex new "drugs" offered by versatile polymer chemistry provides an undeniable advantage for polymer based nano-delivery systems compared to other colloids in this regard. With seven formulations already in different stages of clinical trials, polymeric micelles are in the front line of drug development among different ABC-based nano-carriers. The success in rapid advancement of polymeric micelles from bench to bedside is owed to the rational engineering of core/shell structure so that the polymeric micellar carrier can meet the requirements for optimum delivery of specific drug(s) in certain disease condition(s). The engineering efforts in this regard have mostly been aimed at providing efficient drug loading, micellar stabilization, and sustained and/or site specific drug release. The objective of this review is to provide an update on different engineering strategies employed to achieve optimum polymeric micellar formulations. PMID- 21621571 TI - Focus. PMID- 21621572 TI - Development and validation of an eating norms inventory. Americans' lay-beliefs about appropriate eating. AB - What do American adults believe about what, where, when, how much, and how often it is appropriate to eat? Such normative beliefs originate from family and friends through socialization processes, but they are also influenced by governments, educational institutions, and businesses. Norms therefore provide an important link between the social environment and individual attitudes and behaviors. This paper reports on five studies that identify, develop, and validate measures of normative beliefs about eating. In study 1 we use an inductive method to identify what American adults believe are appropriate or desirable eating behaviors. Studies 2 and 3 are used to purify and assess the discriminant and nomological validity of the proposed set of 18 unidimensional eating norms. Study 4 assesses predictive validity and finds that acting in a norm-consistent fashion is associated with lower Body Mass Index (BMI), and greater body satisfaction and subjective health. Study 5 assesses the underlying social desirability and perceived healthiness of the norms. PMID- 21621573 TI - Ghrelin and food reward: the story of potential underlying substrates. AB - The incidence of obesity is increasing at an alarming rate and this worldwide epidemic represents a significant decrease in life span and quality of life of a large part of the affected population. Therefore an understanding of mechanisms underlying food overconsumption and obesity development is urgent and essential to find potential treatments. Research investigating mechanisms underlying obesity and the control of food intake has recently experienced a major shift in focus, from the brain's hypothalamus to additional important neural circuits controlling emotion, cognition and motivated behavior. Among them, the mesolimbic system, and the changes in reward and motivated behavior for food, emerge as new promising treatment targets. Furthermore, there is also growing appreciation of the impact of peripheral hormones that signal nutrition status to the mesolimbic areas, and especially the only known circulating orexigenic hormone, ghrelin. This review article provides a synthesis of recent evidence concerning the impact of manipulation of ghrelin and its receptor on models of food reward/food motivation behavior and the mesolimbic circuitry. Particular attention is given to the potential neurocircuitry and neurotransmitter systems downstream of ghrelin's effects on food reward. PMID- 21621574 TI - Interaction between glutamate dehydrogenase (GDH) and L-leucine catabolic enzymes: intersecting metabolic pathways. AB - Branched-chain amino acids (BCAAs) catabolism follows sequential reactions and their metabolites intersect with other metabolic pathways. The initial enzymes in BCAA metabolism, the mitochondrial branched-chain aminotransferase (BCATm), which deaminates the BCAAs to branched-chain alpha-keto acids (BCKAs); and the branched chain alpha-keto acid dehydrogenase enzyme complex (BCKDC), which oxidatively decarboxylates the BCKAs, are organized in a supramolecular complex termed metabolon. Glutamate dehydrogenase (GDH1) is found in the metabolon in rat tissues. Bovine GDH1 binds to the pyridoxamine 5'-phosphate (PMP)-form of human BCATm (PMP-BCATm) but not to pyridoxal 5'-phosphate (PLP)-BCATm in vitro. This protein interaction facilitates reamination of the alpha-ketoglutarate (alphaKG) product of the GDH1 oxidative deamination reaction. Human GDH1 appears to act like bovine GDH1 but human GDH2 does not show the same enhancement of BCKDC enzyme activities. Another metabolic enzyme is also found in the metabolon is pyruvate carboxylase (PC). Kinetic results suggest that PC binds to the E1 decarboxylase of BCKDC but does not effect BCAA catabolism. The protein interaction of BCATm and GDH1 promotes regeneration of PLP-BCATm which then binds to BCKDC resulting in channeling of the BCKA products from BCATm first half reaction to E1 and promoting BCAA oxidation and net nitrogen transfer from BCAAs. The cycling of nitrogen through glutamate via the actions of BCATm and GDH1 releases free ammonia. Formation of ammonia may be important for astrocyte glutamine synthesis in the central nervous system. In peripheral tissue association of BCATm and GDH1 would promote BCAA oxidation at physiologically relevant BCAA concentrations. PMID- 21621575 TI - Cost-effectiveness analysis of the 10- and 13-valent pneumococcal conjugate vaccines in Argentina. AB - OBJECTIVE: Since the 10-valent pneumococcal conjugate vaccine (PCV-10) and 13 valent pneumococcal conjugate vaccine (PCV-13) were recently licensed for use in Argentina, both vaccines were evaluated to estimate the costs, health benefits and cost-effectiveness of adding a PCV to the routine child immunization schedule. METHODOLOGY: The integrated TRIVAC vaccine cost-effectiveness model from Pan American Health Organization's ProVac Initiative (Version 1.0.65) was used to assess the health outcomes of 20 successive cohorts from birth to 5 years of age. PCV-10 and PCV-13 were each compared to a scenario assuming no PCV vaccination. A 3+1 (three doses+booster) schedule and a vaccination price of US$ 20.75 per dose was assumed in the base case for both vaccines. RESULTS: Introduction of PCV-13 rather than PCV-10 would increase the number of life years gained (LYG) by at least 10%. The number of LYG (and LYG after adjustment for DALY morbidity weights) was 56,882 (64,252) for PCV-10 compared to 65,038 (71,628) for PCV-13. From the health system perspective, the cost per DALY averted was US$ 8973 and US$ 10,948 for PCV-10 and PCV-13 respectively, and US$ 8546 and US$ 10,510 respectively, after incorporating costs saved by households. When PCV13 was compared to PCV10 directly, the additional benefits of PCV-13 was conferred at a cost of US$ 28,147 per DALY averted. Cost-effectiveness was influenced mainly by vaccine price, serotype replacement, pneumonia mortality and discount rate. CONCLUSION: Routine vaccination against S. pneumoniae in Argentina would be cost-effective with either PCV-10 or PCV-13. PCV-13, with higher coverage of local serotypes, would prevent more cases of pneumonia, invasive pneumococcal disease, sequelae and deaths with a higher number of LYG and DALYs averted, but PCV-10, due its higher impact in the prevention of AOM, would save more costs to the healthcare system. PMID- 21621576 TI - Predominance of nontypeable Haemophilus influenzae in children with otitis media following introduction of a 3+0 pneumococcal conjugate vaccine schedule. AB - In Australia the 7-valent pneumococcal conjugate vaccine (PCV7) is administered at 2, 4 and 6 months of age, with no booster dose. Information on bacterial carriage and the aetiology of recurrent acute otitis media (rAOM) after introduction of PCV7 using the 3+0 schedule is required to evaluate the potential impact of second generation pneumococcal vaccines. We found that 2-4 years after introduction of PCV7 in the National Immunisation Program, nontypeable Haemophilus influenzae (NTHi) was the predominant pathogen isolated from the nasopharynx and middle ear of children with a history of rAOM. Compared with healthy controls (n=81), NTHi and Streptococcus pneumoniae carriage rates were significantly higher in children with a history of rAOM (n=186) (19% vs. 56% p<0.0001 and 26% vs. 41%, p=0.02, respectively). Carriage of PCV7 pneumococcal serotypes was rare, whereas PCV7-related and non-PCV7 serotypes were isolated of 38% of cases and 24% of controls. Serotype 19A was the most common serotype isolated from the nasopharynx and middle ear and accounted for 36% (14/39) of total pneumococcal isolates with reduced susceptibility to cotrimoxazole. Of the 119 children carrying NTHi, 17% of isolates were beta-lactamase positive. The scarcity of PCV7 serotypes in children with and without a history of rAOM indicates that the 3+0 PCV7 schedule is preventing carriage and rAOM from PCV7 serotypes. Introduction of new vaccines in Australia with increased pneumococcal serotype and pathogen coverage, including 19A and NTHi, should decrease the circulation of antibiotic-resistant bacteria and reduce the burden of rAOM. PMID- 21621577 TI - Socioeconomic status, demographics, beliefs and A(H1N1) vaccine uptake in the United States. AB - Early vaccination against influenza viruses is a cost-effective solution to prevent contagion and reduce influenza-related morbidity and mortality. In the face of pandemic viruses, such as the A(H1N1), adequate rates of vaccine uptake play a critical role in containing the spread and effects of the disease. In order to understand the reasons underlying the relatively low 2009-2010 A(H1N1) vaccination rates, we conducted an online survey of 1569 respondents drawn from a nationally representative sample of United States (U.S.) adults age 18, and older. Because prior research suggests that vaccination rates are especially low among some U.S. population subgroups, we oversampled participants from minority ethnic/racial groups and those living under the Federal Poverty Level. Our results show that A(H1N1) vaccine uptake is associated with sociodemographic factors, A(H1N1)-related beliefs and seasonal vaccination. That is, A(H1N1) vaccination is strongly associated with age, urbanicity, perceiving the A(H1N1) vaccine as safe and seasonal flu vaccine uptake. Perceptions of safety and season flu vaccination show the strongest associations with A(H1N1) uptake. The reasons people gave to decline vaccination varied by respondents' sociodemographic group. For example, Black participants were the most likely ethnic/racial group to reported having tried to get the vaccine but found it unavailable. Together, these findings suggest some clear pointers towards strategic public health communication efforts calling for communication campaigns towards audiences segmented by social class, race/ethnicity and beliefs, often what advertisers call "psychodemographics". PMID- 21621578 TI - Nasopharyngeal carriage of Streptococcus pneumoniae in Taiwan before and after the introduction of a conjugate vaccine. AB - BACKGROUND: The heptavalent pneumococcal conjugate vaccine was introduced in Taiwan in October 2005. To evaluate the effect of the vaccination, we conducted an active, prospective, large-scale, long-term, and multicenter study to assess the prevalence of nasopharyngeal Streptococcus pneumoniae carriage in Taiwanese children. METHODS: This study was performed at three tertiary teaching hospitals in northern, central, and southern Taiwan. Questionnaires provided demographic, family/household, and medical history data. Pneumococcal isolates were tested for their susceptibility to various antimicrobial agents and serotypes. In addition, influenza virus and Staphylococcus aureus were recovered from nasopharyngeal and nasal swabs, respectively. RESULTS: Between July 2005 and July 2008, 857 pneumococcal strains were recovered from a total of 6057 children aged >2 months to 5 years (carriage rate, 14.1%). Carriage rates differed geographically and varied with subject age. In a multivariate analysis, having at least one sibling, attendance at day-care centers, a history of otitis media, and history of upper respiratory tract infection in the previous 2 weeks were each associated with a higher risk of pneumococcal colonization of the nasopharynx. Staphylococcus aureus nasal colonization was inversely associated with nasopharyngeal carriage of pneumococcus (p=0.000; odds ratio [OR]: 0.48; 95% CI: 0.39-0.58). Daycare attendance was the only risk factor for carriage of penicillin non-susceptible S. pneumoniae (OR: 2.37; 95% CI: 1.22-4.88). Although vaccination rates rose from 2005 to 2008, no concomitant decrease in S. pneumoniae carriage occurred. The rate of penicillin resistance among S. pneumoniae isolates was 92.8% (using the meningitis criteria). The prevalence of cefotaxime resistance (21.6%) was higher than that of penicillin (6.9%; non-meningitis criteria). Slightly more than half (57.4%) of the isolates belonged to strains covered by the heptavalent pneumococcal conjugate vaccine when both vaccine and vaccine-related serotypes were included. CONCLUSIONS: Although vaccination rates rose from 2005 to 2008, no concomitant decrease occurred in S. pneumoniae carriage. Interaction between S. aureus and S. pneumoniae may influence vaccination efficacy. These findings provide baseline data to further compare pneumococcal carriage rates and antibiotic resistance patterns in Taiwanese children as vaccination rates continue to increase. PMID- 21621579 TI - Effects of thermal treatments during cooking, microwave oven and boiling, on the unconjugated microcystin concentration in muscle of fish (Oreochromis niloticus). AB - Understanding the factors that contribute to the risk from fish consumption is a relevant public health concern due to potential adverse effects of cyanobacterial toxins. The aim of this work was to study the influence of two usual cooking practices, microwave oven and boiling, on the microcystin (MCs) concentration in fish muscle (Tilapia, Oreochromis niloticus) spiked with a stock solution (500 MUL) containing a mixture of three toxins (MC-LR, MC-RR, and MC-YR) (1.5 MUg/mL of each toxin). Two different variables were investigated: time of cooking in the microwaves treatment (1 or 5 min), and way of boiling, "boiled muscle" or "continuously heated muscle". All samples were then lyophilized and MCs were extracted and purified (Oasis HLB cartridge) and quantified by HPLC-MS. Furthermore, the waters in which the samples boiled were also analyzed after their purification. The results suggest a reduction on MC-LR (36%) and MC-YR (24.6%) in samples cooked in the microwave for 5 min. Major changes were found when the fish was cooked by the continuous boiling, with a decrease of 45.0% (MC RR), 56.4% (MC-YR) and 59.3% (MC-LR). More studies are necessary to elucidate the mechanisms involved when aquatic food is submitted to usual cooking practices. PMID- 21621580 TI - Effect of added calcium phosphate on enamel remineralization by fluoride in a randomized controlled in situ trial. AB - Dental products containing calcium phosphate and fluoride are claimed to enhance enamel remineralization over fluoride products. OBJECTIVES: To compare remineralization of enamel subsurface lesions by dental products with added calcium phosphate in a double-blind, randomized, cross-over in situ study. METHODS: Human enamel specimens with subsurface lesions were prepared and inserted into intra-oral appliances worn by volunteers. A slurry (1g product plus 4 ml H(2)O) of each product was rinsed for 60s, 4 times per day for 10 days. Six products were tested (i) placebo, (ii) 1000 ppm F, (iii) 5000 ppm F, (iv) Tooth Mousse (TM), (v) TM plus 900 ppm F (TMP) and (vi) Clinpro with 950 ppm F. Calcium, inorganic phosphate and fluoride levels were measured in post rinse/saliva samples using ion chromatography. Mineral content was measured using transverse microradiography. RESULTS: Only TM and TMP significantly increased salivary calcium and phosphate levels. The products produced remineralization in the following order from lowest to highest: placebo<1000 ppm F=Clinpro<5000 ppm F 0.050) difference was found in the caecal microflora on day 42. Finally, no significant (P > 0.050) difference was noticed on mortality, body and carcass weight. PMID- 21621629 TI - Progressive improvement in cutaneous and extracutaneous chronic graft-versus-host disease after a 24-week course of extracorporeal photopheresis--results of a crossover randomized study. AB - In a prior multicenter randomized controlled trial, we found that a 12-week course of extracorporeal photopheresis (ECP) plus standard immunosuppressive therapy resulted in several beneficial outcomes in patients with corticosteroid refractory/intolerant/dependent chronic graft-versus-host disease (GVHD). Here, we report the results of an open-label crossover ECP study in 29 eligible participants randomized initially to the standard of care non-ECP (control) arm. Eligible for the crossover ECP study were control arm patients who either (1) had progression of cutaneous chronic GVHD (cGVHD), defined as >25% worsening from baseline as measured by the percent change in the total skin score (TSS) at any time, or (2) had less than 15% improvement in the TSS, or had a <=25% reduction in corticosteroid dose at week 12 of the initial study. ECP was administered 3 times during week 1, then twice weekly until week 12, followed by 2 treatments monthly until week 24. The median age of the study cohort was 43 (20-67) years and 90% had extensive cGVHD. The median months from onset of cGVHD to start of ECP were 26 (range: 4-79). Twenty-five of 29 patients (86%) completed the 24-week course of ECP. Complete or partial skin response at week 24 was noted in 9 patients (31%). The median percent of decrease in TSS from baseline to weeks 12 and 24 was -7.9 and -25.8, respectively. In 4 (17%) and 8 (33%) patients, a >=50% reduction in corticosteroid dose at weeks 12 and 24 was observed. Extracutaneous cGVHD response was highest in oral mucosa with 70% complete and partial resolution after week 24. In conclusion, progressive improvement in cutaneous and extracutaneous cGVHD was observed after a 24-week course of ECP in patients who previously had no clinical improvement or exhibited worsening of cGVHD while receiving standard immunosuppressive therapy alone in a randomized study. These results confirm previous findings and support the notion that prolonged ECP appears to be necessary for optimal therapeutic effects in corticosteroid refractory cGVHD patients. PMID- 21621630 TI - Treatment options for transformed lymphoma: incorporating allogeneic stem cell transplantation in a multimodality approach. AB - Transformed non-Hodgkin lymphoma (TL) arising from follicular lymphoma carries a poor prognosis, and the median survival time after transformation is approximately 10 to 12 months. Standard chemotherapy and radioimmunotherapy have offered promising responses; however, the duration of response does not appear to last long. Several studies evaluating the role of autologous stem cell transplantation (auto-SCT) as a salvage regimen have been reported, and a subset of patients benefit from this modality of treatment. With an improvement in supportive care, outcome after allogeneic stem cell transplantation (allo-SCT) has been improved significantly over past decades; however, very limited data are available for TL. In the era of emerging novel therapies, the actual timing, optimal conditioning regimens, and long-term impact of the type of stem cell transplantation (auto-SCT vs allo-SCT) is unclear. This review addresses the approaches to the management of patients with TL. PMID- 21621631 TI - Long noncoding RNAs in cell and developmental biology. PMID- 21621632 TI - The association between fractional exhaled nitric oxide (FeNO) and cat dander in asthmatic children. AB - BACKGROUND: The aim of our study was to assess risk factors of increased FeNO in asthmatic children with no cat at home. METHODS: It was a retrospective, cross sectional study. We evaluated data from medical documentation of children with asthma: FeNO results, allergen sensitization, seasonal allergen exposure, FEV(1), allergic rhinitis (AR) diagnosis and cat presence at home. We assessed asthma severity using mean doses of inhaled glucocorticosteroids and a management approach based on control according to the newest guidelines of Global Initiative for Asthma (GINA) throughout the last three months before the measurement of FeNO and spirometry. RESULTS: 316 patients (age 6-18) completed the study. Sensitization to cat dander was associated with the highest median value of FeNO concentration compared to other allergens in our patients (28,4ppb) and co existing sensitization did not affect FeNO level. Median levels of FeNO increased linearly with patient's age. In asthmatics with AR, the levels of FeNO were increased significantly compared to asthmatics without AR (20.8 vs. 16.3, respectively). We showed that in patients without AR, sensitization to cat allergen was associated with more severe asthma in comparison to other perennial allergy (step 4 vs. other steps according to GINA treatment steps). The above relation was not observed in patients with AR. We did not observe correlation between allergy profile and FEV(1) among patients in neither subgroup nor in general population. CONCLUSIONS: We revealed that sensitization to cat dander was associated with the highest increase of FeNO concentration compared to other allergens in patients not having any cat at home ever. We also observed that in patients without allergic rhinitis, sensitization to cat allergen, compared to other perennial allergy, was associated with more severe asthma. PMID- 21621633 TI - Proinflammatory response of human endothelial cells to Brucella infection. AB - Although vascular pathologies such as vasculitis, endocarditis and mycotic aneurysms have been described in brucellosis patients, the interaction of Brucella with the endothelium has not been characterized. In this study we show that Brucella abortus and Brucella suis can infect and replicate in primary human umbilical vein endothelial cells (HUVEC) and in the microvascular endothelial cell line HMEC-1. Infection led to an increased production of IL-8, MCP-1 and IL 6 in HUVEC and HMEC-1 cells, and an increased expression of adhesion molecules (CD54 in both cells, CD106 and CD62E in HUVEC). Experiments with purified antigens from the bacterial outer membrane revealed that lipoproteins (Omp19) but not lipopolysaccharide mediate these proinflammatory responses. Infection of polarized HMEC-1 cells resulted in an increased capacity of these cells to promote the transmigration of neutrophils from the apical to the basolateral side of the monolayer, and the same phenomenon was observed when the cells were stimulated with live bacteria from the basolateral side. Overall, these results suggest that Brucella spp. can infect and survive within endothelial cells, and can induce a proinflammatory response that might be involved in the vascular manifestations of brucellosis. PMID- 21621634 TI - Mechanotransduction and the regulation of mTORC1 signaling in skeletal muscle. AB - Mechanical stimuli play a major role in the regulation of skeletal muscle mass, and the maintenance of muscle mass contributes significantly to disease prevention and issues associated with the quality of life. Although the link between mechanical signals and the regulation of muscle mass has been recognized for decades, the mechanisms involved in converting mechanical information into the molecular events that control this process remain poorly defined. Nevertheless, our knowledge of these mechanisms is advancing and recent studies have revealed that signaling through a protein kinase called the mammalian target of rapamycin (mTOR) plays a central role in this event. In this review we will, (1) discuss the evidence which implicates mTOR in the mechanical regulation of skeletal muscle mass, (2) provide an overview of the mechanisms through which signaling by mTOR can be regulated, and (3) summarize our current knowledge of the potential mechanisms involved in the mechanical activation of mTOR signaling. PMID- 21621635 TI - Low-dose effects and biphasic effect profiles: is trenbolone a genotoxicant? AB - Over the last years, extensive research has documented endocrine-disrupting activities for a significant number of substances including, among others, hormones, pharmaceuticals, pesticides and surfactants. Nonetheless, for most endocrine disruptors, toxicological profiles are still incomplete or even lacking. A systematic review has shown that a number of endocrine disruptors with steroid-modulating effects may also exert mutagenic and carcinogenic activities. For trenbolone, an androgenic compound, there is controversy about its genotoxic properties in the literature, apparently with a strong dependence on the choice of the test system. Since fish and other aquatic animals are at risk of exposure to run-offs from cattle feedlots or sewage-discharge sites containing trenbolone, potential consequences to aquatic ecosystems need to be assessed. To this end, the potential genotoxic hazard of trenbolone was tested in vitro in the permanent rainbow trout-liver cell-line RTL-W1, as well as in primary cell cultures derived from zebrafish (Danio rerio) embryos after in vivo exposure. In either test system, a potential genotoxic hazard characterized by biphasic dose-response curves could be documented even at exposure concentrations of 30MUg/L. These results thus confirm the conclusion that the steroid trenbolone may act as a genotoxic substance. PMID- 21621636 TI - Single and combined genotoxic and cytotoxic effects of two xenobiotics widely used in intensive aquaculture. AB - Several chemicals are used in aquaculture to prevent or to treat disease outbreaks. These substances are mainly administered by two different routes: by prolonged immersion or by mixing into the diet. In the case of intensive aquaculture, the chemicals that are most frequently applied by immersion are formaldehyde (FA) 37% and oxytetracycline (OTC). The first is highly effective against most protozoa, as well as some of the most common parasites such as monogenetic trematodes. OTC presents a large spectrum of antibacterial activities and is used to treat systemic bacterial infections that affect fish. Under therapeutic use, FA (37%) is applied prophylactically at 200ml/m(3), whereas OTC is used curatively at 40g/m(3). The goal of the present study is to assess genotoxic and cytotoxic effects associated with exposure of the European sea bass (Dicentrarchus labrax) to FA37% and OTC under the same conditions as those applied in intensive aquaculture systems. To this end the micronucleus (MN) assay was applied in erythrocytes. Our results show that both tested chemicals present genotoxic and cytotoxic potential following a time-dependent pattern. Remarkably, the combined treatment induces a cumulative effect, which is particularly pronounced after 15 days of exposure. This suggests the critical hazards associated with exposure to FA and OTC when applied or released together. PMID- 21621637 TI - Maternal-fetal transfer and metabolism of vitamin A and its precursor beta carotene in the developing tissues. AB - The requirement of the developing mammalian embryo for retinoic acid is well established. Retinoic acid, the active form of vitamin A, can be generated from retinol and retinyl ester obtained from food of animal origin, and from carotenoids, mainly beta-carotene, from vegetables and fruits. The mammalian embryo relies on retinol, retinyl ester and beta-carotene circulating in the maternal bloodstream for its supply of vitamin A. The maternal-fetal transfer of retinoids and carotenoids, as well as the metabolism of these compounds in the developing tissues are still poorly understood. The existing knowledge in this field has been summarized in this review in reference to our basic understanding of the transport and metabolism of retinoids and carotenoids in adult tissues. The need for future research on the metabolism of these essential lipophilic nutrients during development is highlighted. This article is part of a Special Issue entitled: Retinoid and Lipid Metabolism. PMID- 21621638 TI - Differential effects of short- and long-term high-fat diet feeding on hepatic fatty acid metabolism in rats. AB - Imbalance in the supply and utilization of fatty acids (FA) is thought to contribute to intrahepatic lipid (IHL) accumulation in obesity. The aim of this study was to determine the time course of changes in the liver capacity to oxidize and store FA in response to high-fat diet (HFD). Adult male Wistar rats were fed either normal chow or HFD for 2.5weeks (short-term) and 25weeks (long term). Short-term HFD feeding led to a 10% higher palmitoyl-l-carnitine-driven ADP-stimulated (state 3) oxygen consumption rate in isolated liver mitochondria indicating up-regulation of beta-oxidation. This adaptation was insufficient to cope with the dietary FA overload, as indicated by accumulation of long-chain acylcarnitines, depletion of free carnitine and increase in FA content in the liver, reflecting IHL accumulation. The latter was confirmed by in vivo((1))H magnetic resonance spectroscopy and Oil Red O staining. Long-term HFD feeding caused further up-regulation of mitochondrial beta-oxidation (24% higher oxygen consumption rate in state 3 with palmitoyl-l-carnitine as substrate) and stimulation of mitochondrial biogenesis as indicated by 62% higher mitochondrial DNA copy number compared to controls. These adaptations were paralleled by a partial restoration of free carnitine levels and a decrease in long-chain acylcarnitine content. Nevertheless, there was a further increase in IHL content, accompanied by accumulation of lipid peroxidation and protein oxidation products. In conclusion, partially effective adaption of hepatic FA metabolism to long-term HFD feeding came at a price of increased oxidative stress, caused by a combination of higher FA oxidation capacity and oversupply of FA. PMID- 21621640 TI - Are biochemical biomarker responses related to physiological performance of juvenile sea bass (Dicentrarchus labrax) and turbot (Scophthalmus maximus) caged in a polluted harbour? AB - Biomarker responses to toxic exposure have been used for decades to indicate stress in aquatic organisms, or the magnitude of environmental pollution. However, little has been done to compare the simultaneous responses of both biochemical and physiological biomarkers. The purpose of this study was twofold. Firstly to analyse the responses of several biochemical biomarkers measured on juvenile sea bass and turbot caged in a northern France harbour at a reference and contaminated stations. Several biotransformation parameters (Ethoxyresorufin O-deethylase - EROD - and Glutathione S-transferase -GST) and an antioxidant enzyme (Catalase -CAT) were analysed. Secondly, to compare their responses to several growth and condition indices, measured on the same fish. In the contaminated station, EROD and GST activities were found to be significantly higher, and a decrease of CAT activity was observed for both species. For individual sea bass, biochemical biomarkers showed numerous significant correlations with growth and condition indices, such as the Fulton's K condition index, the RNA:DNA ratio and the lipid storage index. On the contrary, there were only a few significant correlations for turbot, suggesting a species-specific response. Our study indicates that the analysis of the simultaneous responses of both biochemical and physiological biomarkers can be useful for monitoring complex exposure and to assess habitat quality. PMID- 21621639 TI - Physiological insights into all-trans-retinoic acid biosynthesis. AB - All-trans-retinoic acid (atRA) provides essential support to diverse biological systems and physiological processes. Epithelial differentiation and its relationship to cancer, and embryogenesis have typified intense areas of interest into atRA function. Recently, however, interest in atRA action in the nervous system, the immune system, energy balance and obesity has increased considerably, especially concerning postnatal function. atRA action depends on atRA biosynthesis: defects in retinoid-dependent processes increasingly relate to defects in atRA biogenesis. Considerable evidence indicates that physiological atRA biosynthesis occurs via a regulated process, consisting of a complex interaction of retinoid binding-proteins and retinoid recognizing enzymes. An accrual of biochemical, physiological and genetic data have identified specific functional outcomes for the retinol dehydrogenases, RDH1, RDH10, and DHRS9, as physiological catalysts of the first step in atRA biosynthesis, and for the retinal dehydrogenases RALDH1, RALDH2, and RALDH3, as catalysts of the second and irreversible step. Each of these enzymes associates with explicit biological processes mediated by atRA. Redundancy occurs, but seems limited. Cumulative data support a model of interactions among these enzymes with retinoid binding proteins, with feedback regulation and/or control by atRA via modulating gene expression of multiple participants. The ratio apo-CRBP1/holo-CRBP1 participates by influencing retinol flux into and out of storage as retinyl esters, thereby modulating substrate to support atRA biosynthesis. atRA biosynthesis requires the presence of both an RDH and an RALDH: conversely, absence of one isozyme of either step does not indicate lack of atRA biosynthesis at the site. This article is part of a Special Issue entitled: Retinoid and Lipid Metabolism. PMID- 21621641 TI - Computed tomography enterography detects intestinal wall changes and effects of treatment in patients with Crohn's disease. AB - BACKGROUND & AIMS: The use of computed tomography enterography (CTE) in patients with Crohn's disease has increased. However, there is little data available on how radiologic parameters of active disease change during treatment with infliximab and whether these changes correspond to symptoms, serum biomarkers, or endoscopic appearance. METHODS: We performed a retrospective study of patients with Crohn's disease who had undergone serial CTE imaging while receiving infliximab. Lesions were defined as improved if their enhancement or length decreased without worsening of other parameters. Patients were grouped as responders (all lesions improved), partial responders (some lesions improved), and nonresponders (worsening or no changes in all lesions). Of the 63 patients identified (47% female), the median age was 37.7 years, the median disease duration was 7.6 years, and the median time between initial and first follow-up CTE was 356 days (interquartile range, 215-630). RESULTS: Of 105 lesions, 52 (49.5%) improved, 11 (10.5%) remained unchanged, and 42 (40.0%) worsened. Per patient, 28 (44.4%) were responders, 12 (19.0%) were partial responders, and 23 (36.5%) were nonresponders. The radiologic response had poor-to-fair agreement with symptoms, endoscopic appearance, and levels of C-reactive protein at time of second CTE (kappa = 0.26, 0.07, and 0.30 respectively). CONCLUSIONS: Radiologic improvement was observed in 63.4% of patients with Crohn's disease who received infliximab therapy, despite a study design that was likely biased toward nonresponders. Radiologic response was not in good agreement with clinical symptoms, serum biomarkers, or endoscopic appearance; CTE might be used as a complementary approach to identify mural healing or inflammation not detected by other methods. PMID- 21621642 TI - Gastrojejunal stoma diameter predicts weight regain after Roux-en-Y gastric bypass. PMID- 21621643 TI - Does injection-assisted polypectomy make a difference in assessing resection? PMID- 21621644 TI - Nattectin a fish C-type lectin drives Th1 responses in vivo: licenses macrophages to differentiate into cells exhibiting typical DC function. AB - Considerable efforts are currently focused on the biology of DC in view of their possible clinical use as adjuvant for the generation of antigen-specific immunity and lifelong immunologic memory or for the treatment of tumors. We assessed the role of Nattectin a C-type lectin identified in the Thalassophryne nattereri fish venom in DC maturation. Nattectin induced a significant neutrophilic recruitment into peritoneal cavity of mice, followed by macrophages, with lipidic mediators and IL-12 p70 synthesis. Macrophages derived from 7day-Nattectin mice were CD11c+CD11b(low)Ly6(high)F4/80R(high) and express high levels of MHC class II and CD80 molecules. Culture of peritoneal exudates derived macrophages from 7day Nattectin-mice and immature BMDCs with Nattectin markedly increased the surface expression of CD40, CD80, CD86, and MHC class II in a dose-dependent manner, and the production of MMP-2 and MMP-9 distributed in nucleus and cytoplasm of cells, that was associated with strong activity in the culture supernatant. Nattectin treated DCs secreted IL-12 p70 and IL-10. The Nattectin-treated BMDC or macrophage-derived DCs were highly efficient at Ag capture. The specific immune response elicited by Nattectin was characterized by the production of specific antibodies IgG1 and mainly IgG2a with IL-10 and IFN-gamma synthesis by splenic cells. These results enable us to address that Nattectin induces the recruitment of Ly6C(high) monocytes into the peritoneum, which exhibit a pro-inflammatory profile, where they differentiate into proliferating F4/80R(high) macrophages. Macrophage-derived DCs mature in the presence of the cytokine milieu generated against Nattectin, exhibiting T cell co-stimulatory molecule expression and induced a Th1 polarized response. PMID- 21621645 TI - Nickel allergy-promoting effects of microbial or inflammatory substances at the sensitization step in mice. AB - Microbial components stimulate innate immunity via Toll-like receptors (TLRs), nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs), and/or IL 1. We recently reported that in mice, Escherichia coli lipopolysaccharide (LPS, TLR4-ligand) promotes allergic responses to nickel (Ni) at both the sensitization and elicitation steps. Here, we examined in mice the effects of administering other microbial or inflammatory materials at the Ni-sensitization step. A mixture of 1mM NiCl(2) and a test solution was injected into BALB/c mice intraperitoneally (0.1 ml/10 g body weight), and 10 days later 5mM NiCl(2) was challenged intradermally into the ear pinnas of the mice (20 MUl/ear). The following preparations or substances exhibited adjuvant activities: Prevotella intermedia LPS, Saccharomyces cerevisiae mannan, a synthetic muramyl dipeptide (NOD2-stimulating cell-wall component of bacteria), Pam(3)Cys-SKKKK (TLR2 stimulating synthetic peptide), poly I:C (TLR3-stimulating double-stranded RNA), concanavalin A (a typical T-cell mitogen and T-cell-mediated hepatitis-inducer), heat-killed Propionibacterium acnes (Gram-positive bacterium that causes pimples and induces macrophage-mediated experimental hepatitis), and nitrogen-containing bisphosphonates (chemicals stimulating IL-1 production). Unexpectedly, P. intermedia LPS, which displayed the most potent adjuvant activity among the tested preparations, was effective in TLR4-dysfunctional mutant mice, but not in TLR2-deficient mice, whereas the reverse was true for S. cerevisiae mannan. These results suggest that (i) for the establishment of Ni-allergy in mice, stimulation of innate immunity (including TLRs, NLRs, IL-1 production, and/or other factors) may be important at the sensitization step, and (ii) P. intermedia may produce a substance(s) that potently promotes Ni-allergy via stimulation of TLR2. PMID- 21621646 TI - Honokiol isolated from Magnolia officinalis stimulates osteoblast function and inhibits the release of bone-resorbing mediators. AB - There has been a strong interest in searching for natural therapies for osteoporosis. Honokiol is a phenolic compound isolated from the bark of Magnolia officinalis, a plant widely used in traditional medicine. In the present study, the effects of honokiol on the function of osteoblastic MC3T3-E1 cells were studied. Honokiol caused a significant elevation of cell growth, alkaline phosphatase activity, collagen synthesis, mineralization, glutathione content, and osteoprotegerin release in the cells (P<0.05). Moreover, honokiol significantly (P<0.05) decreased the production of osteoclast differentiation inducing factors such as TNF-alpha, IL-6, and receptor activator of nuclear factor-kB ligand (RANKL) in the presence of antimycin A, which inhibits mitochondrial electron transport and has been used as a ROS generator. These results demonstrate that honokiol may have positive effects on skeletal structure. PMID- 21621647 TI - Arctigenin induces cell cycle arrest by blocking the phosphorylation of Rb via the modulation of cell cycle regulatory proteins in human gastric cancer cells. AB - Gastric cancer is a leading cause of cancer-related deaths, worldwide being second only to lung cancer as a cause of death. Arctigenin, a representative dibenzylbutyrolactone lignan, occurs in a variety of plants. However, the molecular mechanisms of arctigenin for anti-tumor effect on gastric cancer have not been examined. This study examined the biological effects of arctigenin on the human gastric cancer cell line SNU-1 and AGS. Cell proliferation was determined by MTT assay. In MTT assay, the proliferation of SNU-1 and AGS cells was significantly inhibited by arctigenin in a time and dose dependent manner, as compared with SNU-1 and AGS cells cultured in the absence of arctigenin. Inhibition of cell proliferation by arctigenin was in part associated with apoptotic cell death, as shown by changes in the expression ratio of Bcl-2 to Bax by arctigenin. Also, arctigenin blocked cell cycle arrest from G(1) to S phase by regulating the expression of cell cycle regulatory proteins such as Rb, cyclin D1, cyclin E, CDK4, CDK2, p21Waf1/Cip1 and p15 INK4b. The antiproliferative effect of arctigenin on SNU-1 and AGS gastric cancer cells revealed in this study suggests that arctigenin has intriguing potential as a chemopreventive or chemotherapeutic agent. PMID- 21621648 TI - Triptolide protects mice from ischemia/reperfusion injury by inhibition of IL-17 production. AB - Ischemia and reperfusion have been identified as a complex cascade of inflammatory mediators that are involved in the pathogenesis of hepatic injury. Triptolide (diterpenoid triepoxide), was extracted from a purified component of a traditional Chinese Medicine, Tripterygium wilfondii Hook F. Currently, triptolide has been shown to have anti-inflammatory, immunosuppressive, and antineoplastic activity. Accumulated data have shown that Th17 cells might contribute to the pathogenesis of liver diseases. Triptolide has been shown to reduce interleukin (IL)-17 expression in inflammatory bowel disease and arthritis. However, the role of triptolide in liver ischemia/reperfusion (I/R) and whether it can attenuate injury and the potential mechanism have not been investigated. Mice were treated with triptolide (0.1mg/kg) for 1 week or IL-17 antibody (50 MUg/mouse) 2 days before ischemic insult. Partial warm ischemia was produced in the hepatic lobes of C57BL/6 mice for 90 min, followed by various periods of reperfusion. We demonstrated that IL-17 was involved in the inflammatory response to hepatic I/R injury, and that triptolide inhibited IL-17 generation and suppressed neutrophil migration after liver I/R injury through downregulation of signal transducer and activator of transcription 3 (STAT3) transcription. Also, triptolide pretreatment protected the liver from warm I/R injury, at least in part, mediated by the upregulation of Foxp3 expression. These results could pave the way for the use of triptolide as a novel agent to attenuate I/R injury. PMID- 21621649 TI - The uptake of PLGA micro or nanoparticles by macrophages provokes distinct in vitro inflammatory response. AB - Biodegradable micro/nanoparticles generated from PLGA have recently attracted attention due to their clinically proven biocompatibility, especially for immunization purposes. These polymeric particulate delivery systems are able to present antigens and activate both humoral and cellular responses. Many studies have discussed the ideal size of these particles in contributing to the generation of the different types of immune response. However, these studies do not demonstrate the effect of micro or nanoparticles, without any encapsulated bioactive, on phagocytic cells after the uptake process. In this context, the aim of this study was to analyze the in vitro inflammatory behavior of J774 murine macrophages after particles' uptake, since nano/microparticles per se can differently activate phagocytic cells, using or not appropriate receptors, inducing distinct inflammatory responses. An o/w emulsion solvent extraction evaporation method was chosen to prepare the particles. We determined their diameters, zeta potential and morphology. Fluorescent particles' uptake by J774 murine "macrophage-like" cells was also analyzed. To evaluate the in vitro inflammatory profile of these cells after micro or nanoparticles' uptake, we conducted NF-kappaB translocation assay by confocal microscopy and also determined the pro-inflammatory cytokines production provoked by the particles. PMID- 21621650 TI - Effects of different forms of dyspnoea on pain perception induced by cold-pressor test. AB - Although dyspnoea has been shown to attenuate pain, whether different forms of dyspnoea exert a similar inhibitory effect on pain has never been tested. We examined the effects of two different forms of dyspnoea, i.e., "air hunger" sensation (AIR HUNGER) and "work/effort" sensation (WORK/EFFORT), on pain induced by a cold-pressor test. Dyspnoea was induced by two different dyspnoea stimuli (i.e., AIR HUNGER and WORK/EFFORT stimuli) and the magnitudes of both sensations were evaluated by using a visual analogue scale (VAS). At equi-dyspneic VAS levels of two different forms of dyspnoea, pain was induced and the unpleasantness of pain was assessed by pain VAS, pain threshold time (PTT) and pain endurance time (PET). Both AIR HUNGER and WORK/EFFORT caused an increase in PTT and an increase in PET or a decrease in maximal pain VAS. Our findings suggest that AIR HUNGER and WORK/EFFORT exert a similar analgesic effect although the WORK/EFFORT-induced analgesia was slightly more effective. PMID- 21621651 TI - Exercise intolerance at high altitude (5050 m): critical power and W'. AB - The relationship between work rate (WR) and its tolerable duration (t(LIM)) has not been investigated at high altitude (HA). At HA (5050 m) and at sea level (SL), six subjects therefore performed symptom-limited cycle-ergometry: an incremental test (IET) and three constant-WR tests (% of IET WR(max), HA and SL respectively: WR(1) 70+/-8%, 74+/-7%; WR(2) 86+/-14%, 88+/-10%; WR(3) 105+/-13%, 104+/-9%). The power asymptote (CP) and curvature constant (W') of the hyperbolic WR-t(LIM) relationship were reduced at HA compared to SL (CP: 81+/-21 vs. 123+/ 38 W; W': 7.2+/-2.9 vs. 13.1+/-4.3 kJ). HA breathing reserve (estimated maximum voluntary ventilation minus end-exercise ventilation) was also compromised (WR(1): 25+/-25 vs. 50+/-18 l min(-1); WR(2): 4+/-23 vs. 38+/-23 l min(-1); WR(3): -3+/-18 vs. 32+/-24 l min(-1)) with near-maximal dyspnea levels (Borg) (WR(1): 7.2+/-1.2 vs. 4.8+/-1.3; WR(2): 8.8+/-0.8 vs. 5.3+/-1.2; WR(3): 9.3+/-1.0 vs. 5.3+/-1.5). The CP reduction is consistent with a reduced O(2) availability; that of W' with reduced muscle-venous O(2) storage, exacerbated by ventilatory limitation and dyspnea. PMID- 21621652 TI - The peptidases of Trypanosoma cruzi: digestive enzymes, virulence factors, and mediators of autophagy and programmed cell death. AB - Trypanosoma cruzi, the agent of the American Trypanosomiasis, Chagas disease, contains cysteine, serine, threonine, aspartyl and metallo peptidases. The most abundant among these enzymes is cruzipain, a cysteine proteinase expressed as a mixture of isoforms, some of them membrane-bound. The enzyme is an immunodominant antigen in human chronic Chagas disease and seems to be important in the host/parasite relationship. Inhibitors of cruzipain kill the parasite and cure infected mice, thus validating the enzyme as a very promising target for the development of new drugs against the disease. In addition, a 30kDa cathepsin B like enzyme, two metacaspases and two autophagins have been described. Serine peptidases described in the parasite include oligopeptidase B, a member of the prolyl oligopeptidase family involved in Ca(2+)-signaling during mammalian cell invasion; a prolyl endopeptidase (Tc80), against which inhibitors are being developed, and a lysosomal serine carboxypeptidase. Metallopeptidases homologous to the gp63 of Leishmania spp. are present, as well as two metallocarboxypeptidases belonging to the M32 family, previously found only in prokaryotes. The proteasome has properties similar to those of other eukaryotes, and its inhibition by lactacystin blocks some differentiation steps in the life cycle of the parasite. This article is part of a Special Issue entitled: Proteolysis 50 years after the discovery of lysosome. PMID- 21621653 TI - RNA-dependent RNA polymerases from different hepatitis C virus genotypes reveal distinct biochemical properties and drug susceptibilities. AB - The RNA-dependent RNA polymerase of the hepatitis C virus (HCV) is the key enzyme for viral replication, recognized as one of the promising targets for antiviral intervention. Several of the known non-nucleoside HCV polymerase inhibitors (NNIs) identified by screening approaches show limitations in the coverage of all six major HCV genotypes (GTs). Genotypic profiling therefore has to be implemented early in the screening cascade to discover new broadly active NNIs. This implies knowledge of the specific individual biochemical properties of polymerases from all GTs which is to date limited to GT 1 only. This work gives a comprehensive overview of the biochemical properties of HCV polymerases derived from all major GTs 1-6. Biochemical analysis of polymerases from 38 individual sequences revealed that the optima for monovalent cations, pH and temperature were similar between the GTs, whereas significant differences concerning concentration of the preferred cofactor Mg(2+) were identified. Implementing the optimal requirements for the polymerases from each individual GT led to significant improvements in their enzymatic activities. However, the specific activity was distributed unequally across the GTs and could be ranked in the following descending order: 1b, 6a>2a, 3a, 4a, 5a>1a. Furthermore, the optimized assay conditions for genotypic profiling were confirmed by testing the inhibitory activity of 4 known prototype NNIs addressing the NNI binding sites 1 to 4. PMID- 21621654 TI - Effects of monopropanediamino-beta-cyclodextrin on the denaturation process of the hybrid protein BlaPChBD. AB - Irreversible accumulation of protein aggregates represents an important problem both in vivo and in vitro. The aggregation of proteins is of critical importance in a wide variety of biomedical situations, ranging from diseases (such as Alzheimer's and Parkinson's diseases) to the production (e.g. inclusion bodies), stability, storage and delivery of protein drugs. beta-Cyclodextrin (beta-CD) is a circular heptasaccharide characterized by a hydrophilic exterior and a hydrophobic interior ring structure. In this research, we studied the effects of a chemically modified beta-CD (BCD07056), on the aggregating and refolding properties of BlaPChBD, a hybrid protein obtained by inserting the chitin binding domain of the human macrophage chitotriosidase into the class A beta-lactamase BlaP from Bacillus licheniformis 749/I during its thermal denaturation. The results show that BCD07056 strongly increases the refolding yield of BlaPChBD after thermal denaturation and constitutes an excellent additive to stabilize the protein over time at room temperature. Our data suggest that BCD07056 acts early in the denaturation process by preventing the formation of an intermediate which leads to an aggregated state. Finally, the role of beta-CD derivatives on the stability of proteins is discussed. PMID- 21621655 TI - Structural stability of Staphylococcus xylosus lipase is modulated by Zn(2+) ions. AB - Lipases are well-known enzymes extensively used in industrial biotransformation processes. Besides, their structural and catalytic characteristics have attracted increasing attention of several industries in the last years. In this work, we used biophysical and molecular modeling tools to assess structural properties of Staphylococcus xylosus lipase (SXL). We studied the thermal unfolding of this protein and its zinc-dependent thermotolerance. We demonstrated that SXL is able to be active and stable at moderate temperatures, but this feature is only acquired in the presence of Zn(2+). Such characteristic indicates SXL as a zinc dependent metallolipase. PMID- 21621656 TI - Submicron sized ultra-high molecular weight polyethylene wear particle analysis from revised SB Charite III total disc replacements. AB - Submicron sized particles are frequently observed in retrieved total hip and knee periprosthetic tissues and appear to be critical in the activation of the phagocytic inflammatory response. In this paper the concentration, size and shape of ultra-high molecular weight polyethylene (UHMWPE) wear particles between 0.05 and 2.00MUm were determined after isolation from periprosthetic tissues from retrieved lumbar SB Charite III total disc replacements (TDR) using scanning electron microscopy (SEM). For comparison, UHMWPE wear particles were isolated from gamma-radiation-air sterilized total hip arthroplasty (THA) revision tissues. The mean concentration of UHMWPE particles in TDR tissues was 1.6*10(9)g(-1)tissue (range 1.3-2.0), which was significantly lower than the concentration of 2.3*10(9)g(-1) THA revision tissue (range 1.8-3.2) (P=0.03). The mean particle size (equivalent circular diameter: TDR, 0.46MUm; THA 0.53MUm, P=0.60) and mean shape were comparable between TDR and THA (aspect ratio: TDR, 1.89; THA, 1.99, P=0.35; roundness: TDR, 0.58; THA, 0.56, P=0.35). However, the TDR particles tended to be smaller and more round. Although no correlations were found between visible damage to the UHMWPE core and the concentration or shape of the UHMWPE particles, a positive correlation was found between increasing particle size and increasing rim penetration of the TDR core (P=0.04). The presence of UHMWPE particles of similar size and shape in TDR tissue, albeit lower in concentration, might explain why, unlike THA, pain rather than osteolysis is the major reason for revision surgery. PMID- 21621657 TI - Comparative proteomic analysis of Fasciola hepatica juveniles and Schistosoma bovis schistosomula. AB - Protein interactions between host and parasites can influence the infection success and severity. The aim of this investigation was to identify the proteins from two trematodes potentially localized at the host-parasite interface. We performed the proteomic profiles from in vivo obtained immature lung stage Schistosoma bovis schistosomula and in vitro excysted juveniles from Fasciola hepatica, parasites of ruminants and man usually giving rise to chronic infections. Proteomes from those parasites were obtained after digestion with trypsin and the peptides generated were identified by mass spectrometry, both before and after parasites' treatment with 70% methanol. The comparison of the two proteome sets from each parasite and between them, the analysis of their relative abundance and of their potential exposure to the host from living parasites, together with the specific immunolocalization of two of the identified molecules, show that this approach could assist in the identification of parasite exposed proteins and in the definition of molecules common for the two parasites with potential interaction with the host. Further characterization of these molecules could guide to define new common anti-parasitic targets and potential vaccine candidates. PMID- 21621658 TI - Molecular subtypes of osteosarcoma identified by reducing tumor heterogeneity through an interspecies comparative approach. AB - The heterogeneous and chaotic nature of osteosarcoma has confounded accurate molecular classification, prognosis, and prediction for this tumor. The occurrence of spontaneous osteosarcoma is largely confined to humans and dogs. While the clinical features are remarkably similar in both species, the organization of dogs into defined breeds provides a more homogeneous genetic background that may increase the likelihood to uncover molecular subtypes for this complex disease. We thus hypothesized that molecular profiles derived from canine osteosarcoma would aid in molecular subclassification of this disease when applied to humans. To test the hypothesis, we performed genome wide gene expression profiling in a cohort of dogs with osteosarcoma, primarily from high risk breeds. To further reduce inter-sample heterogeneity, we assessed tumor intrinsic properties through use of an extensive panel of osteosarcoma-derived cell lines. We observed strong differential gene expression that segregated samples into two groups with differential survival probabilities. Groupings were characterized by the inversely correlated expression of genes associated with 'G2/M transition and DNA damage checkpoint' and 'microenvironment-interaction' categories. This signature was preserved in data from whole tumor samples of three independent dog osteosarcoma cohorts, with stratification into the two expected groups. Significantly, this restricted signature partially overlapped a previously defined, predictive signature for soft tissue sarcomas, and it unmasked orthologous molecular subtypes and their corresponding natural histories in five independent data sets from human patients with osteosarcoma. Our results indicate that the narrower genetic diversity of dogs can be utilized to group complex human osteosarcoma into biologically and clinically relevant molecular subtypes. This in turn may enhance prognosis and prediction, and identify relevant therapeutic targets. PMID- 21621659 TI - MicroCT morphometry analysis of mouse cancellous bone: intra- and inter-system reproducibility. AB - The agreement between measurements and the relative performance reproducibility among different microcomputed tomography (microCT) systems, especially at voxel sizes close to the limit of the instruments, is not known. To compare this reproducibility 3D morphometric analyses of mouse cancellous bone from distal femoral epiphyses were performed using three different ex vivo microCT systems: GE eXplore Locus SP, Scanco MUCT35 and Skyscan 1172. Scans were completed in triplicate at 12 MUm and 8 MUm voxel sizes and morphometry measurements, from which relative values and dependence on voxel size were examined. Global and individual visually assessed thresholds were compared. Variability from repeated scans at 12 MUm voxel size was also examined. Bone volume fraction and trabecular separation values were similar, while values for relative bone surface, trabecular thickness and number varied significantly across the three systems. The greatest differences were measured in trabecular thickness (up to 236%) and number (up to 218%). The relative dependence of measurements on voxel size was highly variable for the trabecular number (from 0% to 20% relative difference between measurements from 12 MUm and 8 MUm voxel size scans, depending on the system). The intra-system reproducibility of all trabecular measurements was also highly variable across the systems and improved for BV/TV in all the systems when a smaller voxel size was used. It improved using a smaller voxel size in all the other parameters examined for the Scanco system, but not consistently so for the GE or the Skyscan system. Our results indicate trabecular morphometry measurements should not be directly compared across microCT systems. In addition, the conditions, including voxel size, for trabecular morphometry studies in mouse bone should be chosen based on the specific microCT system and the measurements of main interest. PMID- 21621660 TI - Comparing biofeedback with active exercise and passive treatment for the management of work-related neck and shoulder pain: a randomized controlled trial. AB - OBJECTIVES: To compare the effects of biofeedback with those of active exercise and passive treatment in treating work-related neck and shoulder pain. DESIGN: A randomized controlled trial with 3 intervention groups and a control group. SETTING: Participants were recruited from outpatient physiotherapy clinics and a local hospital. PARTICIPANTS: All participants reported consistent neck and shoulder pain related to computer use for more than 3 months in the past year and no severe trauma or serious pathology. A total of 72 potential participants were recruited initially, of whom a smaller group of individuals (n=60) completed the randomized controlled trial. INTERVENTIONS: The 3 interventions were applied for 6 weeks. In the biofeedback group, participants were instructed to use a biofeedback machine on the bilateral upper trapezius (UT) muscles daily while performing computer work. Participants in the exercise group performed a standardized exercise program daily on their own. In the passive treatment group, interferential therapy and hot packs were applied to the participants' necks and shoulders. The control group was given an education booklet on office ergonomics. MAIN OUTCOME MEASURES: Pain (visual analog scale), neck disability index (NDI), and surface electromyography were assessed preintervention and postintervention. Pain and NDI were reassessed after 6 months. RESULTS: Postintervention, average pain and NDI scores were reduced significantly more in the biofeedback group than in the other 3 groups, and this was maintained at 6 months. Cervical erector spinae muscle activity showed significant reductions postintervention in the biofeedback group, and there were consistent trends of reductions in the UT muscle activity. CONCLUSIONS: Six weeks of biofeedback training produced more favorable outcomes in reducing pain and improving muscle activation of neck muscles in patients with work-related neck and shoulder pain. PMID- 21621661 TI - Evaluation of the cost-effectiveness of electrical stimulation therapy for pressure ulcers in spinal cord injury. AB - OBJECTIVE: To evaluate the incremental cost-effectiveness of electrical stimulation (ES) plus standard wound care (SWC) as compared with SWC only in a spinal cord injury (SCI) population with grade III/IV pressure ulcers (PUs) from the public payer perspective. DESIGN: A decision analytic model was constructed for a 1-year time horizon to determine the incremental cost-effectiveness of ES plus SWC to SWC in a cohort of participants with SCI and grade III/IV PUs. Model inputs for clinical probabilities were based on published literature. Model inputs, namely clinical probabilities and direct health system and medical resources were based on a randomized controlled trial of ES plus SWC versus SWC. Costs (Can $) included outpatient (clinic, home care, health professional) and inpatient management (surgery, complications). One way and probabilistic sensitivity (1000 Monte Carlo iterations) analyses were conducted. SETTING: The perspective of this analysis is from a Canadian public health system payer. PARTICIPANTS: Model target population was an SCI cohort with grade III/IV PUs. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Incremental cost per PU healed. RESULTS: ES plus SWC were associated with better outcomes and lower costs. There was a 16.4% increase in the PUs healed and a cost savings of $224 at 1 year. ES plus SWC were thus considered a dominant economic comparator. Probabilistic sensitivity analysis resulted in economic dominance for ES plus SWC in 62%, with another 35% having incremental cost-effectiveness ratios of $50,000 or less per PU healed. The largest driver of the economic model was the percentage of PU healed with ES plus SWC. CONCLUSIONS: The addition of ES to SWC improved healing in grade III/IV PU and reduced costs in an SCI population. PMID- 21621662 TI - Association of muscle strength and bone mineral density in adult survivors of childhood acute lymphoblastic leukemia. AB - OBJECTIVES: To investigate the association between bone mineral density (BMD) and muscle strength in survivors of childhood acute lymphoblastic leukemia (ALL), a population at increased risk for both decreased BMD and muscle strength from cancer and its treatment. DESIGN: Cohort data from the St Jude Lifetime Cohort (SJLIFE) study. SETTING: Department of Cancer Control at St Jude Children's Research Hospital. PARTICIPANTS: Subjects were adults enrolled in St Jude Lifetime Cohort study and treated for childhood ALL between 1962 and 1999. As part of a comprehensive evaluation, participants had dual energy x-ray absorptiometry (DEXA) scans and muscle strength testing. The participants consisted of 261 women and 232 men who were 20.4 to 49.8 years old (median, 35.7y), and 12.7 to 46.5 years from diagnosis of childhood ALL (median, 27.2y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: BMD was determined by DEXA scan. Muscle strength of upper and lower extremities was assessed with physical performance testing. RESULTS: After adjusting for covariates, we found significant (P<0.005) associations between BMD and muscle strength in lower extremities (R(2) range, 0.33-0.40) and strong, significant associations in upper extremities (left-side R(2)=0.558; right-side R(2)=0.560). CONCLUSIONS: Muscle strength was associated with BMD in the extremities of long-term survivors of childhood ALL, a finding suggesting that muscle strengthening interventions may improve bone health in them. PMID- 21621663 TI - Sunday physiotherapy reduces inpatient stay in knee arthroplasty: a retrospective cohort study. AB - OBJECTIVE: To evaluate, in patients treated with knee arthroplasty, whether providing Sunday (7d/wk) physiotherapy intervention could decrease length of stay (LOS) without affecting discharge knee functional outcomes. DESIGN: Retrospective cohort study. SETTING: A tertiary hospital. PARTICIPANTS: Patients (N=229) who underwent total or unicompartmental knee arthroplasty. INTERVENTIONS: In October 2009, we implemented a Sunday physiotherapy intervention program for patients undergoing knee arthroplasty on Friday or Saturday. Prior to this initiative, physiotherapy intervention was provided from Monday to Saturday. LOS and discharge knee outcomes were compared between patients seen during the 4-month intervention period (n=73) and patients seen in the 4 months prior to the intervention (n=82). To control for secular trends, LOS during the intervention period was compared with LOS during the same 4 months in the prior year (n=74). MAIN OUTCOME MEASURES: LOS and discharge knee functional outcomes comprising (1) passive knee flexion range of motion, (2) the proportion of patients who were able to perform an unassisted straight-leg raise, and (3) the proportion of patients who ambulated safely with the use of a single walking stick or quadstick. RESULTS: During the Sunday physiotherapy intervention period, median LOS significantly reduced (5-4d). Controlling for secular trends in LOS revealed similar results. The number needed to treat for 1 additional patient to achieve a LOS 4 days or shorter with Sunday physiotherapy intervention was between 3 and 4. There were no significant changes in knee functional outcomes. CONCLUSIONS: Sunday physiotherapy intervention was associated with a modest reduction in LOS and a faster short-term knee recovery. PMID- 21621664 TI - Effect of 12 weeks of Tai Chi training on soleus Hoffmann reflex and control of static posture in older adults. AB - OBJECTIVE: To investigate the effect of 12 weeks of Tai Chi training on soleus (SOL) Hoffmann reflex (H-reflex) modulation and postural control in standing under 4 sensory conditions in older adults. DESIGN: Experimental research design with pre- and posttraining tests in a training group and a control group. SETTING: University biomechanics laboratory. PARTICIPANTS: Community-dwelling older adults (N=34) were assigned to a training (n=20; mean +/- SD age, 72.9+/ 4.4y) and a control (n=14; mean +/- SD age, 72.9+/-6.5y) group. INTERVENTION: Tai Chi participants attended a 1-hour session of Yang style Tai Chi, 3 sessions a week, for 12 weeks, while control participants maintained their regular daily activities during the same period. MAIN OUTCOME MEASURES: SOL H-reflex (maximal amplitudes of H-reflex [H(max)] and M-wave [M(max)] waves) and mean displacement of the center of pressure (COP) in the anterior-posterior (COP(A-P)) and medial lateral (COP(M-L)) directions were measured during bipedal standing, with the feet placed on a forceplate and the heels 6cm apart, under 4 sensory conditions: stable surface with eyes open, stable surface with eyes closed, unstable surface with eyes open, and unstable surface with eyes closed. RESULTS: SOL H(max)/M(max) ratio in the Tai Chi group was upregulated significantly in all 4 sensory tasks after the 12-week Tai Chi training (P<.05). No significant change in COP measurements (mean displacement of COP(A-P) and COP(M-L)) was found in either the Tai Chi or control group after the 12-week period. CONCLUSIONS: An increase in SOL H(max)/M(max) ratio during static postural tasks is observed after 12 weeks of Tai Chi training in older adults under all 4 sensory conditions. However, training-induced changes in H-reflex were not accompanied by improvement of performance in the static postural control tasks. PMID- 21621665 TI - Wheelchair-related accidents: relationship with wheelchair-using behavior in active community wheelchair users. AB - OBJECTIVE: To report the prevalence, mechanisms, self-perceived causes, consequences, and wheelchair-using behaviors associated with wheelchair-related accidents. DESIGN: A case-control study. SETTING: Community. PARTICIPANTS: A sample of experienced, community-dwelling, active manual and powered wheelchair users (N=95) recruited from a hospital assistive technology service center. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Wheelchair-using behaviors, wheelchair-related accidents over a 3-year period, and the mechanisms and consequences of the accidents. RESULTS: Among the 95 participants, 52 (54.7%) reported at least 1 accident and 16 (16.8%) reported 2 or more accidents during the 3 years prior to the interview. A total of 74 accidents, were categorized into tips and falls (87.8%), accidental contact (6.8%), and dangerous operations (5.4%). A logistic regression found individuals who failed to maintain their wheelchairs regularly (odds ratio [OR]=11.28; 95% confidence interval [CI], 2.62 48.61) and used a wheelchair not prescribed by professionals (OR=4.31; 95% CI, 1.10-16.82) had significantly greater risks of accidents. In addition to the risk factor, lack of regular wheelchair maintenance, the Poisson regression corroborated the other risk factor, seat belts not used (incident rate ratio=2.14; 95% CI, 1.08-4.14), for wheelchair-related accidents. CONCLUSIONS: Wheelchair-related accidents are closely related to their wheelchair-using behaviors. Services including professional evaluation, repair, maintenance, and an educational program on proper wheelchair use may decrease the risks of wheelchair accidents. PMID- 21621666 TI - A 5-year longitudinal study of fatigue in patients with late-onset sequelae of poliomyelitis. AB - OBJECTIVES: To study the severity and 5-year course of fatigue in patients with late-onset sequelae of poliomyelitis (LOSP) and to identify physical and psychosocial determinants of fatigue. DESIGN: Prospective cohort study with 5 measurements over 5 years. SETTING: University hospital. PARTICIPANTS: Patients with LOSP (N=168); 89% of the subjects completed the study. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Fatigue assessed with the Fatigue Severity Scale (FSS). Potential determinants were perceived physical functioning, bodily pain and mental health, extent of paresis, walking capacity, comorbidity, sleeping disorders, coping, and social support. Associations were investigated by multivariable longitudinal analysis using generalized estimating equations. RESULTS: The mean FSS score +/- SD at baseline was 5.1+/-1.4, which did not change significantly during the 5-year follow-up. Reduced physical functioning, increased bodily pain, reduced sleep quality, more psychologic distress, and higher task-oriented coping were independently associated with fatigue. The extent of paresis and walking capacity were strongly associated with physical functioning. CONCLUSIONS: Fatigue is severe and persistent in patients with LOSP due to physical and psychologic factors, which has implications for counseling and treatment. In addition to the commonly applied interventions targeting physical aspects, psychologic interventions are a potential area for reducing fatigue. PMID- 21621667 TI - Mobility assessment: sensitivity and specificity of measurement sets in older adults. AB - OBJECTIVE: To identify quantitative measurement variables that characterize mobility in older adults, meet reliability and validity criteria, distinguish fall risk, and predict future falls. DESIGN: Observational study with 1-year weekly falls follow-up. SETTING: Mobility laboratory. PARTICIPANTS: Community dwelling volunteers (N=74; age, 65-94y) categorized at entry as 27 nonfallers or 47 fallers by using Medicare criteria (1 injury fall or >1 noninjury fall in the previous year). INTERVENTIONS: None. MAIN OUTCOME MEASURES: Test-retest and within-subject reliability, criterion and concurrent validity; predictive ability indicated by observed sensitivity and specificity to entry fall-risk group (falls status), Tinetti Performance Oriented Mobility Assessment (POMA), computerized dynamic posturography Sensory Organization Test (SOT), and subsequent falls reported weekly. RESULTS: Measurement variables were selected that met reliability (intraclass coefficient of correlation >.6) and/or discrimination (P<.01) criteria (clinical variables: turn steps and time, gait velocity, step-in tub time, downstairs time; forceplate variables: quiet standing Romberg ratio sway area, maximal lean anterior-posterior excursion, sit-to-stand medial-lateral excursion, sway area). Sets were created (3 clinical, 2 forceplate) using combinations of variables appropriate for older adults with different functional activity levels, and composite scores were calculated. Scores identified entry falls status and concurred with POMA and SOT scores. The full clinical set (5 measurement variables) produced sensitivity of 80% and specificity of 74% to falls status. Composite scores were more sensitive and specific overall in predicting subsequent injury falls and multiple falls compared with falls status and POMA or SOT score. CONCLUSIONS: Sets of quantitative measurement variables obtained with this mobility battery provided sensitive prediction of future injury falls and screening for multiple subsequent falls by using tasks that should be appropriate to diverse participants. PMID- 21621668 TI - Low interrater reliability of examiners performing the prone instability test: a clinical test for lumbar shear instability. AB - OBJECTIVE: To test the interrater reliability of examiners performing the prone instability test (PIT), a clinical test proposed to identify lumbar shear instability. DESIGN: Cross-sectional test-retest design examining individuals with mechanical low back pain (LBP). SETTING: University-based musculoskeletal analysis laboratory. PARTICIPANTS: Individuals (N=30) with mechanical LBP recruited from community sources in a metropolitan region. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Repeated measures of a clinical examination test proposed to identify lumbar shear instability. RESULTS: Interrater reliability of examiners' judgments of PIT results were indexed with percentage of agreement and kappa statistic. Examiners obtained 63% agreement and kappa of .10 (95% confidence interval, -.27 to .47). Adjusted kappa values based on prevalence and bias indexes were calculated to evaluate the effect on kappa. The prevalence index associated with examiner judgments of the PIT was .43, and bias index was .03. The prevalence-adjusted bias-adjusted kappa value was slightly higher than the unadjusted kappa value (kappa=.27; 95% confidence interval, -.08 to .61). CONCLUSIONS: Results of our study are not consistent with those of previous studies examining the reliability of therapists performing the PIT. We conclude that examiners do not attain acceptable interrater reliability when performing procedures for the PIT based on the information currently provided in the literature. Based on our experience, we suggest further exploration, standardization, and clarification of procedural details to improve therapists' ability to conduct the PIT on individuals with LBP. PMID- 21621669 TI - Invited commentary on "low interrater reliability of examiners performing the prone instability test: a clinical test for lumbar shear instability". AB - The stated purpose of the article by Ravenna and colleagues is to examine the reliability of the prone instability test (PIT) in the hands of novice clinicians, as previous reliability studies have primarily included more experienced clinicians. They conclude that the interrater reliability for the PIT is low when performing the test as described in the current literature; however, their conclusions should be interpreted cautiously given the significant modifications made to standardize the PIT in their study, as these changes greatly limit the capacity to make direct comparisons to earlier reliability studies of the PIT. The modifications to the PIT used in this study are discussed, including the use of a stabilization belt, alternate positioning of the hips and knees, and the use of a stool for foot placement. PMID- 21621670 TI - Electrophysiologic evidence of spinal accessory neuropathy in patients with cervical myofascial pain syndrome. AB - OBJECTIVE: To evaluate whether or not spinal accessory neuropathy exists in patients with cervical myofascial pain syndrome (MFPS). DESIGN: Prospective study. SETTING: A neurophysiologic laboratory in a university hospital. PARTICIPANTS: Patients with cervical MFPS (n=25) and healthy controls (n=20). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: We performed nerve conduction studies (NCSs) in bilateral spinal accessory nerves, and electromyography and stimulated single-fiber electromyography in the trapezius muscles of all patients and controls. Parameters including nerve conduction velocities (NCVs), amplitudes and areas of compound muscle action potentials (CMAPs), and mean consecutive differences (MCDs) in single-fiber electromyography were measured, analyzed, and compared with the disease durations of the patients. RESULTS: Spinal accessory NCSs showed normative NCVs but with prominently reduced CMAP amplitude in the patients with cervical MFPS, which is recognized as an axonal neuropathy of the spinal accessory nerves. Electromyography showed prominent evidence of denervation and reinnervation patterns in 48% of the MFPS patients. The abnormal MCDs in single-fiber electromyography indicated a synaptic delay of motor endplates in the motor units, and may signify evolving instability of neuromuscular transmission in the spinal accessory nerves innervating trapezius muscles of the patients. CONCLUSIONS: This study demonstrates electrophysiologic evidence of neuroaxonal degeneration and neuromuscular transmission disorder in a significant proportion of patients with cervical MFPS. We suggest that spinal accessory neuropathy may be associated with cervical MFPS. PMID- 21621671 TI - Variation in results of volume measurements of stumps of lower-limb amputees: a comparison of 4 methods. AB - OBJECTIVE: To analyze the reliability of 4 methods (water immersion, computer aided design [CAD] photometric method, CAD hand scanner, and circumferential measurements) for stump volume measurement in transtibial amputees. DESIGN: Repeated measurements. SETTING: General community, ambulatory care. PARTICIPANTS: Transtibial amputees (N=26; mean age +/- SD, 58.7+/-11.0y). INTERVENTIONS: Stump volume of patients with an amputation was measured on 2 occasions, each consisting of 2 sessions. In each session, stump volume was measured by 2 observers using each of the 4 methods. Sequence of observers and measurement methods was determined randomly. MAIN OUTCOME MEASURE: Repeatability coefficients, as a measure for reliability, for each method were calculated, as well as variance components to estimate the influence of measurement conditions on stump volumes measured. RESULTS: Repeatability coefficients varied from 129mL CAD hand scanner to 158mL CAD photometric method. Error variance contributed 12% to the total variance. Methods contributed 36%, method-amputee and occasion amputee interactions contributed both 25% to the error variance. CONCLUSIONS: Repeatability coefficient was lowest for the CAD hand scanner, which indicates the best reliability. Substantial differences existed in stump volumes measured between the 4 methods. PMID- 21621672 TI - Association between exercise and fear of falling in community-dwelling elderly Koreans: results of a cross-sectional public opinion survey. AB - OBJECTIVE: To investigate the association between habitual exercise and fear of falling in an older Korean population. DESIGN: Cross-sectional study conducted in a population-based sample of an urban city. SETTING: Urban city in South Korea. PARTICIPANTS: Randomly sampled older Korean people (N=828; aged >=65y) living in a typical urban city located in South Korea. INTERVENTION: Standardized telephone interview. MAIN OUTCOME MEASURES: Data on exercise habits, history of falls during the previous year, and fear of falling were obtained using a random digit dialing telephone survey method. Multiple linear regression and multiple logistic regression analyses were used to examine the association between exercise habits and the fear of falling. RESULTS: The incidence of falls in the study cohort was 13%, and the prevalence of the fear of falling was 67.4% (47.6% in men and 80.8% in women). About 30% of those that expressed a fear of falling stated that this was the cause of their limited activity. Older men and women who exercised regularly showed a similar level of fear of falling, but they were less likely to experience fear-related activity restriction than nonexercisers. CONCLUSIONS: A regular exercise habit was found to be inversely associated with fear-related activity restriction regardless of fall experience among older Korean men and women. PMID- 21621673 TI - Physiological responses to body weight--supported treadmill exercise in healthy adults. AB - OBJECTIVE: To determine whether the relationships of heart rate, rating of perceived exertion (RPE), and ground reaction forces (GRFs) with oxygen consumption rate (Vo(2)) during treadmill exercise are altered by partial body weight support (BWS) via lower-body positive pressure. DESIGN: Repeated-measures design. SETTING: Exercise physiology laboratory. PARTICIPANTS: Healthy, active adults (N=12); mean age +/- SD, 45.1+/-12.6 years. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Vo(2), heart rate, RPE, and GRFs were measured during walking and running at 3 levels (0%, 25%, 50%) of BWS. Before exercise, standing heart rate and blood pressure were measured under each BWS condition. RESULTS: Standing heart rates were 7 beats/min lower (P<.05) and systolic blood pressures were 10mmHg higher (P<.001) at 50% BWS compared with 0% BWS, but mean blood pressure while standing and the relationship of heart rate with Vo(2) during walking and running were not altered by BWS. While walking, the RPE at a Vo(2) of 10 mL . kg(-1) . min(-1) was statistically lower (P<.05) at 0% BWS compared with 25% and 50% BWS (mean values, 7 vs 8 points), but RPE was not different among conditions while running at a Vo(2) of 25 mL . kg(-1) . min(-1). Peak normal GRFs at specified Vo(2) levels and RPE values were reduced (P<.05) with increasing BWS for walking and running. CONCLUSIONS: Because partial BWS does not alter the relationship of heart rate with Vo(2) during exercise and has minimal effect on the relationship of RPE with Vo(2), training heart rate and RPE values do not appear to require adjustment with partial BWS. Reduced GRFs at specified Vo(2) levels from partial BWS suggest that there are important clinical applications of this technology. PMID- 21621674 TI - Modified constraint-induced movement therapy versus traditional rehabilitation in patients with upper-extremity dysfunction after stroke: a systematic review and meta-analysis. AB - OBJECTIVE: To compare the effectiveness of modified constraint-induced movement therapy (CIMT) with traditional rehabilitation (TR) therapy in patients with upper-extremity dysfunction after stroke. DATE SOURCES: Computerized database searches and hand-searches, as 2 main search strategies, were used to collect studies. A comprehensive search of PubMed, EMBASE, the Cochrane Library, the Chinese academic journals full-text database, the Chinese biomedical literature database, the Chinese scientific journals database, and Chinese medical association journals was conducted. Relevant journals and article reference lists were hand-searched. Meanwhile, we searched unpublished trials by using the System for Information on Gray Literature database. STUDY SELECTION: Randomized controlled trials (RCTs) only about modified CIMT versus TR for treatment of patients with upper-extremity dysfunction after stroke were identified in this systematic review. Participants included adults age over 18 years with a clinical diagnosis of stroke and met the inclusion criteria of modified CIMT. DATE EXTRACTION: Two reviewers extracted relevant information from included studies according to a date extraction form. The methodologic quality of the included studies was assessed using a quality-scoring instrument, which was a 5-point scale that included a description of randomization, double-blind structure, and withdrawals/dropouts. DATA SYNTHESIS: Thirteen RCTs involving 278 patients (modified CIMT/TR=143/135) were included. Meta-analysis showed that patients receiving modified CIMT showed higher scores for the Fugl Meyer Assessment (mean difference [MD]=7.8; 95% confidence interval [CI], 4.21-11.38), the Action Research Arm Test (MD=14.15; 95% CI, 10.71-17.59), the FIM (MD=7.00; 95% CI, .75 13.26), and the Motor Activity Log: Amount of Use (MD=.78; 95% CI, .37-1.19) and Quality of Use (MD=.84; 95% CI, .42-1.25) than patients in the TR group. In kinematic variables, patients receiving modified CIMT had a shorter reaction time and a higher percentage of movement time where peak velocity occurred than patients receiving TR (P<.05), while meta-analysis showed that there was no significant difference in normalized movement time (P=.99), normalized total displacement (P=.44), and normalized movement unit (P=.68). CONCLUSIONS: This systematic review provided fairly strong evidence that modified CIMT could reduce the level of disability, improve the ability to use the paretic upper extremity, and enhance spontaneity during movement time, but evidence is still limited about the effectiveness of modified CIMT in kinematic analysis. PMID- 21621675 TI - A systematic review of instruments assessing participation: challenges in defining participation. AB - OBJECTIVES: To evaluate: (1) whether instruments which intend to measure participation actually do and (2) how frequently specific aspects and domains of participation are addressed. DATA SOURCES: A systematic search was performed in PubMed. STUDY SELECTION: Included were patient-reported instruments that primarily aim to measure participation. DATA EXTRACTION: The full-text instruments were extracted from the articles or obtained from the authors. Two reviewers independently rated each item of the included instruments as measuring participation (yes, no, or undetermined). For each item, the specific aspect and domain of participation were categorized. DATA SYNTHESIS: Included were 103 instruments (2445 items). Of the included items, 619 items concerned participation and 217 concerned undetermined items. In total, 68 instruments contained at least 1 (sub)scale with 50% or more participation or undetermined items. The participation items referred to the participation aspects: participation problems (53%), participation accomplishment (31%), and satisfaction with participation (9%). The domains of the participation items concerned: work/study (27%), social life (27%), general participation (19%), and home (11%). The undetermined items mainly referred to domains about leisure (43%), transport (26%), and shopping (12%). CONCLUSIONS: According to our working definition of participation, most instruments that aim to measure participation do so only to a limited extent. These instruments mainly assess aspects of participation problems and participation accomplishment. The domains of participation covered by these instruments primarily include work/study, social life, general participation, home, leisure, transport, and shopping. PMID- 21621676 TI - Review of accelerometry for determining daily activity among elderly patients. AB - OBJECTIVES: To review studies that used accelerometers to classify human movements and to appraise their potential to determine the activities of older patients in hospital settings. DATA SOURCES: MEDLINE, CINAHL, and Web of Science electronic databases. A search constraint of articles published in English language between January 1980 and March 2010 was applied. STUDY SELECTION: All studies that validated the use of accelerometers to classify human postural movements and mobility were included. Studies included participants from any age group. All types of accelerometers were included. Outcome measures criteria explored within the studies were comparisons of derived classifications of postural movements and mobility against those made by using observations. Based on these criteria, 54 studies were selected for detailed review from 526 initially identified studies. DATA EXTRACTION: Data were extracted by the first author and included characteristics of study participants, accelerometers used, body positions of device attachment, study setting, duration, methods, results, and limitations of the validation studies. DATA SYNTHESIS: The accelerometer based monitoring technique was investigated predominantly on a small sample of healthy adult participants in a laboratory setting. Most studies applied multiple accelerometers on the sternum, wrists, thighs, and shanks of participants. Most studies collected validation data while participants performed a predefined standardized activity protocol. CONCLUSIONS: Accelerometer devices have the potential to monitor human movements continuously to determine postural movements and mobility for the assessment of functional ability. Future studies should focus on long-term monitoring of free daily activity of a large sample of mobility-impaired or older hospitalized patients, who are at risk for functional decline. Use of a single waist-mounted triaxial accelerometer would be the most practical and useful option. PMID- 21621678 TI - Educational programs for chronic fatigue syndrome need to take cognizance of the condition's abnormal response to exercise. PMID- 21621679 TI - In memoriam: Theodore M. Cole, MD. PMID- 21621680 TI - No more! Organized rape in the Democratic Republic of the Congo must stop now. PMID- 21621681 TI - Methodology and tools for quality improvement in maternal and newborn health care. AB - OBJECTIVE: To gain an overview of approaches, methodologies, and tools used in quality improvement of maternal and newborn health in low-income countries. METHODS: Electronic search of MEDLINE and organizational databases for literature describing approaches, methodologies, and tools used to improve the quality of maternal and newborn health care in low-income countries. Relevant papers and reports were reviewed and summarized. RESULTS: Developing a culture of quality is an important requisite for successful quality improvement. Methodologies to improve quality include the development of standards and guidelines and the performance of mortality, near-miss, and criterion-based audits. Tools for data collection and process description were identified, and examples of work to improve quality of care are provided. CONCLUSION: The documented experience with the identified approaches, methodologies, and tools indicates that none is sufficient by itself to achieve a desirable improvement in quality of care. The choice of methodologies and tools depends on the healthcare system and its available resources. There is a lack of studies that describe the process of quality improvement and a need for research to provide evidence of the effectiveness of the identified methods and tools. PMID- 21621682 TI - [Editorial. Cerebellar diseases]. PMID- 21621683 TI - The case against BRCA 1 and 2 testing. PMID- 21621684 TI - N-acetyl-l-cysteine decreases intra-abdominal adhesion formation through the upregulation of peritoneal fibrinolytic activity and antioxidant defenses. AB - BACKGROUND: Intraperitoneal adhesions occur in more than 94% of patients after abdominal surgery. Mechanisms that decrease oxidative stress and upregulate peritoneal fibrinolysis reduce adhesions. N-acetyl-l-cysteine (NAC) is a clinically relevant antioxidant whose effect on peritoneal fibrinolysis and ability to decrease adhesions has not been established. The aims of this study were to determine if NAC reduces adhesions and to characterize its potential mechanism(s) of action. METHODS: Male Wistar rats (n = 92) received 0.9% saline (OP Control), intraperitoneal NAC (150 mg/kg, OP + NAC), or oral NAC (1200 mg/kg) twice daily on preoperative day 1, day of operation, and postoperative day 1. Adhesions were induced on the day of operation using our previously described ischemic button model. Animals were killed on postoperative day 7 for adhesion scoring. Peritoneal tissue and fluid from the intraperitoneal NAC group were measured at 24 hours for fibrinolytic activity, tissue plasminogen activator (tPA), plasminogen activator inhibitor-1 (PAI-1), total glutathione, and 8 isoprostane (8-IP). The effect of NAC on tPA and PAI-1 production was tested in vitro in human mesothelial cells. The effect of NAC on intestinal wound healing was measured using colonic anastomotic burst pressures. RESULTS: Intraperitoneal NAC reduced adhesions by 53% (P < .001) compared to OP Controls without affecting anastomotic wound healing. NAC increased the tPA/PAI-1 protein ratio and peritoneal fibrinolytic activity by 69% and 127%, respectively, compared to OP Controls (P < .05). NAC did not restore total glutathione levels in peritoneal adhesion tissue but decreased 8-IP by 46% and 65% (P < .05) in peritoneal tissue and fluid, respectively, compared to OP Controls. Human mesothelial cells incubated with NAC exhibited a concentration-dependent increase in the tPA/PAI-1 ratio, which supported in vivo observations (P < .05). Oral NAC did not decrease adhesions. CONCLUSION: NAC administered intraperitoneally decreased adhesion formation while upregulating peritoneal fibrinolytic activity and antioxidant defenses without affecting normal anastomotic wound healing. These data suggest a potential new therapeutic use for NAC in adhesion prevention. PMID- 21621685 TI - Response to "Long-term outcome of 'prophylactic therapy' for familial medullary thyroid cancer (MTC)". PMID- 21621686 TI - Omentoplasty for preventing anastomotic leaks after esophagogastrostomy. PMID- 21621687 TI - The need for standardized technique in intraoperative monitoring of the external branch of the superior laryngeal nerve during thyroidectomy. PMID- 21621688 TI - Re: "Outcomes of elective abdominal aortic aneurysm repair among the elderly: endovascular versus open repair". PMID- 21621689 TI - Nutrition and metabolism of the surgical patient, part I. PMID- 21621690 TI - Nutrition and metabolism of the surgical patient, part I. PMID- 21621691 TI - Metabolic considerations in management of surgical patients. AB - Metabolic changes after surgery, trauma, or serious illness have a complex pathophysiology. The early posttraumatic stress response is physiologic and associated with a state of hyperinflammation, increased oxygen consumption, and increased energy expenditure. These are part of a systemic reaction that encompasses a wide range of endocrinological, immunologic, and hematological effects. Surgery initiates changes in metabolism that can affect virtually all organs and tissues; the metabolic response results in hormone-mediated mobilization of endogenous substrates that leads to stress catabolism. Hypercatabolism has been associated with severe complications related to hyperglycemia, hypoproteinemia, and immunosuppression. Proper metabolic support is essential to restore homeostasis and ensure survival. PMID- 21621692 TI - Management of enterocutaneous fistulas. AB - Management of enterocutaneous fistulas (ECFs) involves (1) recognition and stabilization, (2) anatomic definition and decision, and (3) definitive operation. Phase 1 encompasses correction of fluid and electrolyte imbalance, skin protection, and nutritional support. Abdominal imaging defines the anatomy of the fistula in phase 2. ECFs that do not heal spontaneously require segmental resection of the bowel segment communicating with the fistula and restoration of intestinal continuity in phase 3. The enteroatmospheric fistula (EAF) is a malevolent condition requiring prolonged wound care and nutritional support. Complex abdominal wall reconstruction immediately following fistula resection is necessary for all EAFs. PMID- 21621693 TI - Current management of the short bowel syndrome. AB - Short bowel syndrome is a challenging clinical problem that benefits from a multidisciplinary approach. Much progress has recently been made in all aspects of management. Medical intestinal rehabilitation should be the initial treatment focus, and several new potential pharmacologic agents are being investigated. Surgical rehabilitation using nontransplant procedures in selected patients may further improve intestinal function. Intestinal lengthening procedures are particularly promising. Intestinal transplantation has increasingly been used with improving success in patients with life-threatening complications of intestinal failure. PMID- 21621694 TI - Nutrition support in the pediatric surgical patient. AB - This article deals with the nutritional needs of pediatric patients. It begins by discussing the caloric requirements of different pediatric patients and moves on to a breakdown of the specific nutrients required. It then progresses to a detailed description of the enteral and parenteral modalities for delivery of nutrition to pediatric patients. The article concludes with a discussion of specific problems and disorders encountered in pediatric surgical patients. PMID- 21621695 TI - The prevention and treatment of intestinal failure-associated liver disease in neonates and children. AB - Intestinal Failure Associated Liver Disease (IFALD) is a common and potentially life-threatening problem for pediatric patients receiving long-term parenteral nutrition (PN). Risk factors for IFALD include premature birth, low birth weight, long-term PN, intestinal stasis and sepsis. Preventative strategies are the cornerstone of improving outcomes in IFALD and include enteral feeding, weaning of PN, reduced dose lipid emulsions and the early recognition and treatment of sepsis. Recent work also demonstrates the efficacy of fish-oil based lipid emulsions in the prevention and treatment of IFALD. Transplantation is an option for end-stage liver disease but is associated with significant morbidity and mortality. PMID- 21621696 TI - Adjuvant nutrition management of patients with liver failure, including transplant. AB - This article reviews nutrition support in patients with liver disease, including those who are undergoing surgery or liver transplant. The topics covered include the multifactorial etiology of malnutrition, nutritional assessment, and nutritional therapy. Recommendations for use of both enteral and parenteral nutrition are given in patients with alcoholic hepatitis, cirrhosis, and acute liver failure and in patients undergoing surgery or liver transplant. PMID- 21621697 TI - Nutritional therapy in critically ill and injured patients. AB - Nutritional support of critically ill or injured patients has undergone significant advances in the last few decades. These advances are the direct result of the growing scientific progress and increased knowledge of the biology and biochemistry of key metabolic and nutrient changes induced by injury, sepsis, and other critical illnesses, both in adults and children. As this knowledge has increased, the science of nutritional support has become more disease based and disorder based. This article discusses protein and nitrogen metabolism in critically ill patients, immunomodulation, and the key nutrients involved in an immune-enhancing diet. PMID- 21621698 TI - Nutrition in critical care. AB - Critical care has evolved from a prolonged recovery room stay for cardiac surgery patients to a full medical and nursing specialty in the last 5 decades. The ability to feed patients who cannot eat has evolved from impossible to routine clinical practice in the last 4 decades. Nutrition in critically ill patients based on measurement of metabolism has evolved from a research activity to clinical practice in the last 3 decades. The authors have been involved in this evolution and this article discusses past, present, and likely future practices in nutrition in critically ill patients. PMID- 21621700 TI - Nutrition management of patients with malignancies of the head and neck. AB - The importance of nutrition and the prognostic impact of malnutrition in patients with head and neck cancer are not fully appreciated in the surgical world where a pervasive attitude exists that weight loss during treatment is inevitable and nutritional expertise or intervention may be dismissed out of ignorance. In this article, the authors explore the nutritional requirements of these patients and the impact of a multidisciplinary therapeutic approach to head, neck, and skull base cancer care. PMID- 21621701 TI - Nutritional support: we have failed in our ability to support patients with sepsis and cancer. AB - Despite the success of both parenteral and enteral nutrition in supporting patients who cannot eat, patients with either sepsis or cancer cannot be adequately supported. A proposed mechanism by which aerobic glycolysis leads to a shortage of energy production in the liver is discussed. According to this hypothesis, the proximity of sodium-potassium ATPase and glycogen, its fuel source, leads to the continuation of gluconeogenesis with continued proteolysis and muscle wasting. Myostatin and lipokine, newly discovered factors, may also play a role. PMID- 21621703 TI - Parenteral nutrition and nutritional support of surgical patients: reflections, controversies, and challenges. AB - An abridged background of the history, cause, and nature of early controversies related to the development and clinical application of total parenteral nutrition (TPN) is presented, followed by discussions of highlights of some of the more prominent controversies that continue to prevail, including nutritional support of geriatric patients with and without cancer, the importance of maintenance of normal glycemia, the primacy of nutrition support teams, and TPN versus total enteral nutrition. Reflections related to nutritional support and TPN are interspersed throughout the discussions, together with a compilation of legacies of TPN to the modern practice of medicine. PMID- 21621702 TI - Cachexia, malnutrition, the refeeding syndrome, and lessons from Goldilocks. AB - Cachexia has plagued clinicians for centuries. Although all cachexia is related to malnutrition, cachexia associated with malignant diseases differs from starvation cachexia in that it is more recalcitrant to nutritional therapy. All cachexia responds to judicious nutritional support; however, cancer cachexia worsens autonomously as the disease advances and cannot be arrested or reversed by any known form of nutrition, hormonal, or pharmacologic therapy. Cachexia must be treated cautiously to avoid overfeeding syndrome, which may result in serious or dangerous complications or death. PMID- 21621704 TI - Historical highlights of the development of total parenteral nutrition. AB - The events and discoveries thought to be the most significant prerequisites to the development of total parenteral nutrition (TPN) dating back to the early 17th century are chronicled. A more detailed description and discussion of the subsequent early modern highlights of the basic and clinical research beginning in the mid-20th century and the advances culminating in the first demonstration of the feasibility and practicality of TPN, and its successful, safe and efficacious applications clinically, are presented. Some of the reasoning, insights, and philosophy of a pioneer clinician-scientist in the field are shared with readers. PMID- 21621699 TI - What, how, and how much should patients with burns be fed? AB - The hypermetabolic response to severe burn injury is characterized by hyperdynamic circulation and profound metabolic, physiologic, catabolic, and immune system derangements. Failure to satisfy overwhelming energy and protein requirements after, and during, severe burn injury results in multiorgan dysfunction, increased susceptibility to infection, and death. Attenuation of the hypermetabolic response by various pharmacologic modalities is emerging as an essential component of the management of patients with severe burn injury. This review focuses on the more recent advances in therapeutic strategies to attenuate the hypermetabolic response and its postburn-associated insulin resistance. PMID- 21621705 TI - Base two? PMID- 21621706 TI - Neuroendocrine tumors of the gynecologic tract: A Society of Gynecologic Oncology (SGO) clinical document. AB - OBJECTIVE: Neuroendocrine tumors of the gynecologic tract are rare, and pose a significant clinical challenge because of the tumor heterogeneity and lack of standardized guidelines for treatment. This manuscript summarizes the available literature concerning these tumors in an effort to provide the clinician a framework from which to guide patient management. METHODS: MEDLINE was searched for all research articles published in English between January 1, 1966 and March 1, 2011 in which the studied population included women diagnosed with neuroendocrine tumors of the gynecologic tract. Although preference was given to prospective studies, studies were not limited by design or by numbers of subjects given the limited availability of reports. RESULTS: Most, but not all, neuroendocrine tumors of the gynecologic tract have an aggressive clinical course and those of the cervix histologically and clinically share similarities with small cell lung cancer. Cumulative data supports a multi-modality therapeutic strategy. A proposed management algorithm for neuroendocrine carcinomas of the cervix is outlined. For less frequent disease sites including the adnexa, uterus, vagina and vulva, as well as well differentiated carcinoid tumors, surgical resection is appropriate in selected cases. Etoposide/platinum based chemotherapy is used for neuroendocrine carcinomas but not for well differentiated carcinoid tumors. Well differentiated carcinoid and atypical carcinoid tumors should be managed similar to gastroenteropancreatic NETs (GEP-NETs). CONCLUSIONS: Most neuroendocrine tumors of the gynecologic tract require a multi-modality therapeutic approach, determined by extent of disease and primary organ of involvement. Pathologic diagnosis is critical to guide therapy. PMID- 21621707 TI - The making of a modern medic. PMID- 21621708 TI - Half a century of Amnesty International. PMID- 21621709 TI - Salvage with erlotinib plus bevacizumab: not in NSCLC. PMID- 21621710 TI - Fetal growth standards: individual and global perspectives. PMID- 21621711 TI - The first flight of DSM-5. PMID- 21621713 TI - Keryn Christiansen: a household name in microbiology. PMID- 21621714 TI - The IHP+: a welcome initiative with an uncertain future. PMID- 21621715 TI - A crucial factor in shared decision making: the team approach. PMID- 21621716 TI - Efficacy of bevacizumab plus erlotinib versus erlotinib alone in advanced non small-cell lung cancer after failure of standard first-line chemotherapy (BeTa): a double-blind, placebo-controlled, phase 3 trial. AB - BACKGROUND: Bevacizumab and erlotinib target different tumour growth pathways with little overlap in their toxic-effect profiles. On the basis of promising results from a phase 1/2 trial assessing safety and activity of erlotinib plus bevacizumab for recurrent or refractory non-small-cell lung cancer (NSCLC), we aimed to assess efficacy and safety of this combination in a phase 3 trial. METHODS: In our double-blind, placebo-controlled, randomised phase 3 trial (BeTa), we enrolled patients with recurrent or refractory NSCLC who presented to 177 study sites in 12 countries after failure of first-line treatment. Patients were randomly allocated in a one-to-one ratio to receive erlotinib plus bevacizumab (bevacizumab group) or erlotinib plus placebo (control group) according to a computer-generated randomisation sequence by use of an interactive voice response system. The primary endpoint was overall survival in all enrolled patients. Patients, study staff, and investigators were masked to treatment assignment. We assessed safety by calculation of incidence of adverse events and tissue was collected for biomarker analyses. This trial is registered with ClinicalTrials.gov, number NCT00130728. FINDINGS: Overall survival did not differ between 317 controls and 319 patients in the bevacizumab group (hazard ratio [HR] 0.97, 95% CI 0.80-1.18, p=0.7583). Median overall survival was 9.3 months (IQR 4.1-21.6) for patients in the bevacizumab group compared with 9.2 months (3.8 20.2) for controls. Progression-free survival seemed to be longer in the bevacizumab group (3.4 months [1.4-8.4]) than in the control group (1.7 months [1.3-4.1]; HR 0.62, 95% CI 0.52-0.75) and objective response rate suggested some clinical activity of bevacizumab and erlotinib. However, these secondary endpoint differences could not be defined as significant because the study prespecified that the primary endpoint had to be significant before testing of secondary endpoints could be done, to control type I error rate. In the bevacizumab group, 130 (42%) of 313 patients with safety data had a serious adverse event, compared with 114 (36%) controls. There were 20 (6%) grade 5 adverse events, including two arterial thromboembolic events, in the bevacizumab group, and 14 (4%) in the control group. INTERPRETATION: Addition of bevacizumab to erlotinib does not improve survival in patients with recurrent or refractory NSCLC. FUNDING: Genentech. PMID- 21621717 TI - A global reference for fetal-weight and birthweight percentiles. AB - BACKGROUND: Definition of small for gestational age in various populations worldwide remains a challenge. References based on birthweight are deficient for preterm births, those derived from ultrasound estimates might not be applicable to all populations, and the individualised reference can be too complex to use in developing countries. Our aim was to create a generic reference for fetal weight and birthweight that overcame these deficiencies and could be readily adapted to local populations. METHODS: We used the fetal-weight reference developed by Hadlock and colleagues and the notion of proportionality proposed by Gardosi and colleagues and made the weight reference easily adjustable according to the mean birthweight at 40 weeks of gestation for any local population. For application and validation, we used data from 24 countries in Africa, Latin America, and Asia that participated in the 2004-08 WHO Global Survey on Maternal and Perinatal Health (237,025 births). We compared our reference with that of Hadlock and colleagues (non-customised) and with that of Gardosi and colleagues (individualised). For every reference, the odds ratio (OR) of adverse perinatal outcomes (stillbirths, neonatal deaths, referral to higher-level or special care unit, or Apgar score lower than 7 at 5 min) for infants who were small for gestational age versus those who were not was estimated with multilevel logistic regression. FINDINGS: OR of adverse outcomes for infants small for gestational age versus those not small for gestational age was 1.59 (95% CI 1.53-1.66) for the non-customised fetal-weight reference compared with 2.87 (2.73-3.01) for our country-specific reference, and 2.84 (2.71-2.99) for the fully individualised reference. INTERPRETATION: Our generic reference for fetal-weight and birthweight percentiles can be easily adapted to local populations. It has a better ability to predict adverse perinatal outcomes than has the non-customised fetal-weight reference, and is simpler to use than the individualised reference without loss of predictive ability. FUNDING: None. PMID- 21621718 TI - PAS-positive macrophages--not always infection. PMID- 21621719 TI - Infective endocarditis: the European viewpoint. Foreword. PMID- 21621720 TI - Infective endocarditis: the European viewpoint. AB - Infective endocarditis (IE) is a difficult and complex disease. In recent years epidemiology and microbiology have changed. In developed countries IE is now affecting older patients and patients with no previously known valve disease. Prosthetic IE (prosthetic valve endocarditis [PVE]) and endocarditis in patients with pacemakers and other devices (cardiac device related infective endocarditis [CDRIE]) are becoming more frequent. The number of Staphylococcus aureus IE is increasing related to the number of endocarditis that occurs because of health care associated procedures, especially in diabetics or patients on chronic hemodialysis. The change in the underlying population and the increase in the number of cases caused by very virulent organism explain why the disease still carries a poor prognosis and a high mortality. The variety of clinical manifestations and complications, as well as the serious prognosis, makes it mandatory that IE patients need to be treated in experienced hospitals with a collaborative approach between different specialists, involving cardiologists, infectious disease specialists, microbiologists, surgeons, and frequently others, including neurologists and radiologists. Only an early diagnosis followed by risk stratification and a prompt institution of the correct antibiotic treatment as well as an appropriate and timed surgical indication may improve mortality figures. The recent European Guidelines try to provide clear and simple recommendations, obtained by expert consensus after thorough review of the available literature to all specialists involved in clinical decision-making of this difficult and changing disease. PMID- 21621721 TI - Development of disease-specific quality indicators for Danish chiropractic patients with low back pain. AB - OBJECTIVE: The purpose of this study is to develop disease-specific quality indicators for Danish chiropractic patients with low back pain (LBP) as an initial effort to include chiropractors in the Danish Health Care Quality Programme. METHODS: A cross-disciplinary Indicator Group consisting of researchers and clinicians prioritized 9 evidence-based quality indicators and quality performance standards in a systematic consensus process. A feasibility study involving 20 chiropractors in 8 chiropractic clinics and 206 LBP patients was undertaken. Afterward, an audit meeting was held where participating clinicians could voice their opinion and share experiences. After the test and audit, the Indicator Group reconvened and decided on the final set of indicators. RESULTS: The following quality indicators were chosen: case history, test for discogenic back pain, neurology, radiology, classification, exercise therapy, outcome assessment, and reevaluation. Only the outcome assessment indicators met the standards set by the Indicator Group. Based on the feedback supplied at the audit meeting and after evaluating the test results, the Indicator Group decided to reduce the standards but keep all indicators. CONCLUSIONS: This study showed that it is possible to describe quality indicators for the assessment and treatment of LBP patients, to identify the patient population, and to collect data. The participating clinics did not meet the original quality standards set by the Indicator Group. Reasons for this may include unrealistically high thresholds set by the Indicator Group and the somewhat complex or compound structure of some of the indicators. The Indicator Group and the test clinics recommend future nationwide implementation of the developed quality indicators. PMID- 21621722 TI - Immediate effects on electromyographic activity and pressure pain thresholds after a cervical manipulation in mechanical neck pain: a randomized controlled trial. AB - OBJECTIVE: The purpose of this study was to identify the immediate effects of a manipulation of C5/C6 level on electromyography (EMG) of the deltoid muscle and in pressure pain thresholds (PPTs) in patients with mechanical neck pain. METHODS: Thirty-seven subjects with mechanical neck pain were randomly divided into 2 groups: manipulative group, which received a cervical spine manipulation targeted to C5/C6 segment, and a control group, which did not receive any procedure. Outcomes were EMG data of the deltoid muscle (rest, isometric contraction for 5 or 30 seconds, and isotonic contraction) and PPT over upper trapezius and deltoid muscles and C5 spinous process. They were assessed before and 5 minutes after treatment by a blinded assessor. A 3-way repeated-measures analysis of variance was used to examine the effects of the manipulation. RESULTS: A significant group time interaction for MF at the beginning of isometric contraction for 30 seconds (F = 7.957, P = .006) was also found: the manipulative group experienced a greater increase in MF at the beginning of the isometric contraction than did the control group. A significant group time interaction was also found for root mean square during isometric contraction for 30 seconds (P = .003); however, changes were small. Patients within the manipulative group experienced an increase on PPT over the deltoid (P = .010) and C5 spinous process (P = .025), but not over upper trapezius (P = .776). CONCLUSIONS: Manipulation at C5/C6 level in the study participants seemed to increase EMG amplitude signal and fatigue resistance in a nonspinal (deltoid) muscle innervated by the same segment in patients with mechanical neck pain. However, these changes were relative small. An increase on PPT over those tissues innervated by the manipulated segment was also found after the manipulative procedure. PMID- 21621723 TI - Kinematic analysis of the lumbar spine by digital videofluoroscopy in 18 asymptomatic subjects and 9 patients with herniated nucleus pulposus. AB - OBJECTIVES: The purpose of this study was to use digital videofluoroscopy to identify motion patterns of the lumbar spine during coronal movement in asymptomatic (normal) subjects and patients with herniated nucleus pulposus (HNP). METHODS: Videofluoroscopic lumbar coronal motion was recorded in 18 asymptomatic volunteers and 9 patients with HNP. Measurements were made while patients bent laterally and rotated toward the right and left from a sitting position and then returned to their original position. Direction and degree of extension in the coronal plane at each motion segment and sacral descent were measured. Through the motion analysis software, the coupled pattern with lateral bending and rotation was analyzed in the asymptomatic subjects and patients with HNP. RESULTS: Lateral flexion movement was coupled with contralateral extension and ipsilateral sacral descent but with a different rotation pattern. Rotation movement was coupled with ipsilateral extension, ipsilateral sacral descent, and ipsilateral spinous process rotation. Patients with HNP and asymptomatic subjects had similar coupled patterns but differences in amount of motion. CONCLUSIONS: Digital videofluoroscopy showed coupled patterns during the lateral bending and rotation movements. PMID- 21621724 TI - Validity of the straight-leg raise test for patients with sciatic pain with or without lumbar pain using magnetic resonance imaging results as a reference standard. AB - OBJECTIVE: The aim of this retrospective study was to assess validity of the straight-leg raise (SLR) test using magnetic resonance imaging (MRI) results as a reference standard in a group of patients with L4-L5 and L5-S1 lumbar-herniated disks and sciatic pain. The relationship between diagnostic accuracy of this test, age classes, and grade of lumbar disk displacement was investigated. METHODS: The charts of 2352 patients with sciatic pain with/without lumbar pain were examined. Results of the SLR were then compared with previous spinal MRI. A 2 * 2 contingency table was created, and analysis of sensitivity, specificity, positive and negative predictive values, diagnostic odds ratio, likelihood ratio (LR), and receiver operating characteristic (ROC) curve was carried out. Homogeneous age classes were created to compare them statistically. RESULTS: Magnetic resonance imaging findings showed lumbar disk herniation (LDH) in 1305 patients. Of these subjects, 741 were positive on SLR testing. Sensitivity was 0.36, whereas specificity was 0.74. Positive and negative predictive values were 0.69 and 0.52, respectively. Positive LR was 1.38, and negative LR was 0.87. Diagnostic odds ratio was 1.59, and ROC analysis showed an area under the curve (AUC) of 0.596. The AUC decreased from 0.730 in the 16- to 25-year subgroup to 0.515 in the 76- to 85-year subgroup. Similar results were obtained in subjects with LDH and nerve root compression. CONCLUSIONS: Our results indicate low accuracy of the SLR in diagnosis of LDH if compared with MRI results. The discriminative power of the SLR seemed to decrease as age increased; thus, positive and negative results may be less conclusive in older patients. PMID- 21621725 TI - Interexaminer reliability of supine leg checks for discriminating leg-length inequality. AB - OBJECTIVE: The purpose of this study was to quantify interexaminer reliability of a standardized supine leg check procedure used to screen for leg-length inequality. METHODS: Two doctors of chiropractic used a standardized supine leg check procedure to examine 50 volunteers for leg-length inequality. The order of examination was randomized. The side and magnitude of leg-length inequality were determined to the nearest 1/8 in. Subjects and examiners were blinded. Interexaminer reliability was assessed with a Bland-Altman plot, tolerance table of absolute differences, a quadratic weighted kappa statistic for quantitative scores, and a Gwet's first-order agreement coefficient for dichotomous ratings. RESULTS: The quadratic weighted kappa statistic to quantify the reliability of the rating scale was 0.44 (95% confidence interval, 0.21-0.67), indicating moderate reliability. The 2 examiners agreed exactly 32% of the time, within 1/8 in 58% of the time, within 3/16 in 72% of the time, and within 3/8 in 92% of the time. The Bland-Altman plot revealed possible heterogeneity in reliability that requires additional study. The examiners agreed on the presence of a leg-length inequality of at least 1/8 in in 40 (80%) of 50 subjects (first-order agreement coefficient, 0.76), suggesting good agreement for this diagnostic category. CONCLUSION: The examiners showed moderate reliability in assessing leg-length inequality at 1/8-in increments and good reliability in determining the presence of a leg-length inequality. PMID- 21621726 TI - Interrater reliability of the craniocervical flexion test in asymptomatic individuals--a cross-sectional study. AB - OBJECTIVE: The objective of this study was to evaluate interrater reliability of the craniocervical flexion test (CCFT) on asymptomatic subjects. METHODS: A cross sectional repeated-measures study design was used. Thirty asymptomatic subjects (15 men and 15 women; mean age, 33.7 years; range, 22-48 years) were recruited for the study. Subjects were positioned in supine lying with a pneumatic pressure sensor of the pressure biofeedback unit placed under the neck. Subjects performed 3 trials of craniocervical flexion with each trial consisting of 5 incremental stages (22, 24, 26, 28, and 30 mm Hg) guided through feedback from the pressure dial of the pressure biofeedback unit. All the trials were scored simultaneously by 2 raters. The outcome measure was the activation score-the maximum pressure (above baseline 20 mm Hg) that was achieved and held in a steady manner for 10 seconds. Intraclass correlation coefficient (ICC 3,1) was analyzed using the 2 repeated scores out of 3 trials for either rater. RESULTS: Interrater reliability (ICC) for the CCFT was 0.91 (95% confidence interval, 0.83-0.96). There was a reasonable agreement on the Bland-Altman plot confirming high reliability of the test. CONCLUSION: The study has shown high interrater reliability when 2 raters simultaneously scored the CCFT trials in asymptomatic individuals. PMID- 21621727 TI - Shoe orthotics for the treatment of chronic low back pain: a randomized controlled pilot study. AB - OBJECTIVES: The purpose of this pilot study was to investigate the feasibility of a randomized clinical trial of shoe orthotics for chronic low back pain. METHODS: The study recruited 50 patients with chronic low back pain through media advertising in a midwestern suburban area. Medical history and a low back examination were completed at a chiropractic clinic. Subjects were randomized to either a treatment group receiving custom-made shoe orthotics or a wait-list control group. After 6 weeks, the wait-list control group also received custom made orthotics. This study measured change in perceived pain levels (Visual Analog Scale) and functional health status (Oswestry Disability Index) in patients with chronic low back pain at the end of 6 weeks of orthotic treatment compared with no treatment and at the end of 12 weeks of orthotic treatment. RESULTS: This study showed changes in back pain and disability with the use of shoe orthotics for 6 weeks compared with a wait-list control group. It appears that improvement was maintained through the 12-week visit, but the subjects did not continue to improve during this time. CONCLUSIONS: This pilot study showed that the measurement of shoe orthotics to reduce low back pain and discomfort after 6 weeks of use is feasible. A larger clinical trial is needed to verify these results. PMID- 21621728 TI - Methodological quality of studies on the measurement properties of neck pain and disability questionnaires: a systematic review. AB - OBJECTIVE: The aim of this study was to obtain an overview of the methodological quality of studies on the measurement properties of neck pain and disability questionnaires and to describe how well various aspects of the design and statistical analyses of studies on measurement properties are performed. METHODS: A systematic review was performed of published studies on the measurement properties of neck pain and disability questionnaires. Two reviewers independently rated the quality of the studies using the COnsensus-based Standards for the selection of health Measurement INstruments (COSMIN) checklist. This checklist was developed in an international Delphi consensus study. RESULTS: A total of 47 articles were included on the measurement properties of 8 different questionnaires. The methodological quality of the included studies was adequate on some aspects (often, adequate statistical analyses are used for assessing reliability, measurement error, and construct validity) but can be improved on other aspects. The most important methodological aspects that need to be improved are as follows: assessing unidimensionality in internal consistency analysis, stable patients and similar test conditions in studies on reliability and measurement error, and more emphasis on the relevance and comprehensiveness of the items in content validity studies. Furthermore, it is recommended that studies on construct validity and responsiveness should be based on predefined hypotheses and that better statistical methods should be used in responsiveness studies. CONCLUSION: Considering the importance of adequate measurement properties, it is concluded that, in the field of measuring neck pain and disability, there is room for improvement in the methodological quality of studies measurement properties. PMID- 21621729 TI - New nurses' experience of their role within interprofessional health care teams in mental health. AB - This qualitative study explored new nurses' experience of their role within interprofessional health care teams in a mental health organization in Canada. Semistructured interviews were conducted with 10 nurses. Content analysis revealed two main themes, namely, adopting a passive role to learn how to fit in and engaging in an active role to impact on patient care. Establishing credibility and building trust were central to the new nurses' transition from a passive to a more active role. Interpersonal and organizational factors contributed to the transition. Recommendations for creating healthy work environments that promote interprofessional collaboration and facilitate new nurses' transition into interprofessional health care teams are presented. PMID- 21621730 TI - Relational dimensions of a positive integration experience for new mental health nurses. AB - This study investigated the key aspects of a successful integration experience into mental health nursing from the perspectives of new nurses working in a psychiatric institution. Analysis of 10 semistructured interviews revealed that the relational atmosphere in which new nurses find themselves is determinant of their satisfaction with their integration experience. Quality relational connections with patients constitute the primary motivation of new nurses in choosing a career in mental health. At the same time, quality relational connections between and among team members sustain such motivation. Conversely, a negative relational atmosphere sets the stage for new nurses to experience disillusionment and despair. PMID- 21621731 TI - Postnatal depression in first-time mothers: prevalence and relationships between functional and structural social support at 6 and 12 weeks postpartum. AB - Postnatal depression (PND) is a significant public health issue, with variable prevalence and a dearth of research on risk and protective factors. This quantitative longitudinal study of 512 first-time mothers identified the prevalence of PND and examined the relationships between functional and structural social support at 6 and 12 weeks postpartum. The prevalence of PND was 13.2% at 6 weeks and 9.8% at 12 weeks. At 6 and 12 weeks, the only social support dimension independently associated with PND was total functional social support. At-birth formal structural support and emotional functional support were independently predictive of PND at 12 weeks. PMID- 21621732 TI - Psychological harassment in the nursing workplace: an observational study. AB - Psychological harassment in the workplace involves disrespectful or humiliating behavior to workers. Nurses make up one of the groups that are most exposed to these behaviors. This descriptive study investigated the most common types of psychological harassment in the nursing workplace and their relationship with sociodemographic variables among 285 nurses in Spain. Findings indicate differences in the prevalence of psychological harassment depending on the criterion that was used. Psychological harassment is positively correlated with a desire to abandon the profession and negatively with participation in decision making. The results suggest combining different measures to evaluate psychological harassment in the workplace and zero-tolerance polices for psychological abuse. PMID- 21621733 TI - Buddhist group therapy for diabetes patients with depressive symptoms. AB - The objective of this study was to assess the effect of Buddhist group therapy on patients with type 2 diabetes who had depressive symptoms. A quasi-experimental design study using a control group with matching technique was conducted. After informed consent was obtained, the "Nine questions for assessing depressive disorder symptom" (Isan language) was used to determine the patient's condition. A total of 62 patients with type 2 diabetes who had depressive symptoms were assigned to either the experimental group (n = 32) or the control group (n = 32). Patients in the experimental group were divided further into four groups (8 patients per group) and attended the Buddhist group therapy. The intervention consisted of a weekly Buddhist group gathering lasting 2 hours for 6 weeks plus home meditation practices. Patients in the control group received treatment as usual. Both groups received standard physician treatment, including medication. Physicians did not know who was in either the control or experimental groups. Results show that 6 months after the intervention, 65.6% and 100% of patients in the control group and experimental group, respectively, returned to normal level. The intention-to-treat analysis, which included two participants in the experimental group lost follow-up, yielded a small reduction in the number of patients who returned to normal level (93.8%). With intention-to-treat analysis, the relative risk on depressive symptoms between the experimental and control groups was 6.5 (95% confidence interval, 1.4-30.6). Qualitative data from the experimental group supported that there were therapeutic group factors involved. However, patients realized the truth of being oneself and also accepted their current living condition. In conclusion, this program is effective in reducing depressive symptoms. PMID- 21621734 TI - A longitudinal study of family conflicts, social support, and antenatal depressive symptoms among Chinese women. AB - Little is known about the causal factors of antenatal depressive symptomatology in the Chinese population. A total of 1,527 pregnant women were recruited to investigate the predictors of antenatal depressive symptoms using a stress process model in a prospective longitudinal study. The Edinburgh Postnatal Depression Scale, the Dyadic Adjustment Scale, the Stryker Adjustment Checklist, and the Interpersonal Support Evaluation List were used. Multiple linear regression analysis revealed that marital conflict, parent-in-law conflict, and social support predicted antenatal depressive symptoms. Psychiatric nurses can identify predictors to help initiate preventive intervention. PMID- 21621735 TI - Marriage and mental health: when a spouse has Alzheimer's disease. AB - When spouses suffer from memory disorders, caregivers are at high risk for negative mental health consequences. The purpose of this pilot study was to identify factors within marital relationships that increase risk for caregiver burden and depression. PARTICIPANTS/SAMPLING: Caregivers (5 men and 11 women) and their partners with Alzheimer's disease were English speaking, age 55 and older and living in the community. Caregivers were cognitively intact and spouses had moderate cognitive impairment. METHODS: This was a descriptive correlational study. Martial quality was rated by caregivers (CG) using the Marital Quality Index. Caregiver burden was measured by the Zarit Burden Interview and depression was measured by the 20-item Center for Epidemiological Studies- Depression Scale (CES-D). RESULTS: Spouses who rated their marriage as strong and satisfying experienced less burden than spouses who rated their marriage more negatively (r = -.464, p <.01). IMPLICATIONS: Individuals with Alzheimer's disease are often cared for by spouse caregivers who may become overburdened and depressed. Findings suggest that spouses who were inexperienced caregivers were most vulnerable to negative outcomes and therefore a population warranting closer scrutiny. PMID- 21621737 TI - Targeted inhibitors for steroid transforming enzymes. PMID- 21621738 TI - Forensic medicine department of Dubai. PMID- 21621739 TI - Expression of the human TSPY gene in the brains of transgenic mice suggests a potential role of this Y chromosome gene in neural functions. AB - The testis specific protein Y-encoded (TSPY) is a member of TSPY/SET/NAP1 superfamily, encoded within the gonadoblastoma locus on the Y chromosome. TSPY shares a highly conserved SET/NAP-domain responsible for protein--protein interaction among TSPY/SET/NAP1 proteins. Accumulating data, so far, support the role of TSPY as the gonadoblastoma gene, involved in germ cell tumorigenesis. The X-chromosome homolog of TSPY, TSPX is expressed in various tissues at both fetal and adult stages, including the brain, and is capable of interacting with the multi-domain adapter protein CASK, thereby influencing the synaptic and transcriptional functions and developmental regulation of CASK in the brain and other neural tissues. Similar to TSPX, we demonstrated that TSPY could interact with CASK at its SET/NAP-domain in cultured cells. Transgenic mice harboring a human TSPY gene and flanking sequences showed specific expression of the human TSPY transgene in both testis and brain. The neural expression pattern of the human TSPY gene overlapped with those of the endogenous mouse Cask and Tspx gene. Similarly with TSPX, TSPY was co-localized with CASK in neuronal axon fibers in the brain, suggesting a potential role(s) of TSPY in development and/or physiology of the nervous system. PMID- 21621740 TI - Pioglitazone suppresses advanced glycation end product-induced expression of plasminogen activator inhibitor-1 in vascular smooth muscle cells. AB - Advanced glycation end products (AGEs) play an important role in vascular complications of diabetes, including fibrinolytic abnormalities. Pioglitazone, a peroxisome proliferator-activated receptor gamma (PPARgamma) agonist, has recently been shown to reduce circulating plasminogen activator inhibitor-1 (PAI 1) levels in diabetes mellitus. In the present study, we investigated the effects of pioglitazone on the expression of local PAI-1 in rat vascular smooth muscle cells (VSMCs) induced by AGEs and the underlying mechanism. The result showed that AGEs could enhance the PAI-1 expression by 5.1-fold in mRNA and 2.7-fold in protein level, as evaluated by real-time RT-PCR and Western blotting, respectively. Pioglitazone was found to down-regulate the AGE-stimulated PAI-1 expression in VSMCs. However, these inhibitory effects were partially attenuated by the PPARgamma antagonist, GW9662. Furthermore, we found that AGEs induced a rapid increase in phosphorylation and activation of extracellular signal regulated protein kinase 1/2 (ERK1/2). The ERK kinase inhibitor, U0126, partially prevented the induction of PAI-1 by AGEs. Moreover, pioglitazone was also found to inhibit the phosphorylation of ERK1/2. Taken together, it was concluded that pioglitazone could inhibit AGE-induced PAI-1 expression, which was mediated by the ERK1/2 and PPARgamma pathways. Our findings suggested pioglitazone had a therapeutic potential in improving fibrinolytic activity, and consequently preventing thromboembolic complications of diabetes and cardiovascular disease. PMID- 21621741 TI - Coevolution study of mitochondria respiratory chain proteins: toward the understanding of protein--protein interaction. AB - Coevolution can be seen as the interdependency between evolutionary histories. In the context of protein evolution, functional correlation proteins are ever present coordinated evolutionary characters without disruption of organismal integrity. As to complex system, there are two forms of protein--protein interactions in vivo, which refer to inter-complex interaction and intra-complex interaction. In this paper, we studied the difference of coevolution characters between inter-complex interaction and intra-complex interaction using "Mirror tree" method on the respiratory chain (RC) proteins. We divided the correlation coefficients of every pairwise RC proteins into two groups corresponding to the binary protein--protein interaction in intra-complex and the binary protein- protein interaction in inter-complex, respectively. A dramatical discrepancy is detected between the coevolution characters of the two sets of protein interactions (Wilcoxon test, p-value = 4.4 * 10(-6)). Our finding reveals some critical information on coevolutionary study and assists the mechanical investigation of protein--protein interaction. Furthermore, the results also provide some unique clue for supramolecular organization of protein complexes in the mitochondrial inner membrane. More detailed binding sites map and genome information of nuclear encoded RC proteins will be extraordinary valuable for the further mitochondria dynamics study. PMID- 21621742 TI - A Pid3 allele from rice cultivar Gumei2 confers resistance to Magnaporthe oryzae. AB - Rice blast, caused by Magnaporthe oryzae, is one of the most devastating diseases. Using map-based strategy and in silico approach we isolated a new rice (Oryza sativa L.) blast resistance allele of Pid3, designated Pi25, from a stable blast resistance cultivar Gumei2. Over-expression analysis and complementation test showed that Pi25 conferred blast resistance to M. oryzae isolate js001-20. Sequence analysis showed that Pi25 was an intronless gene of 2,772 nucleotides with single nucleotide substitution in comparison to Pid3 at the nucleotide position 459 and predicatively encoded a typical coiled coil--nucleotide binding site--leucine rich repeat (CC--NBS--LRR) protein of 924 amino acid residuals with 100% identity to Pid3 putative protein. The susceptible allele pi25 in Nipponbare contained a nonsense mutation at the nucleotide position 2,209 resulting in a truncated protein with 736 amino acid residuals. In addition, 14 nucleotide substitutions resulting in 10 amino acid substitutions were identified between Pi25 and pi25 upstream the premature stop codon in the susceptible allele. Although the mechanism of Pi25/Pid3-mediated resistance needs to be further investigated, the isolation of the allele would facilitate the utilization of Pi25/Pid3 in rice blast resistance breeding program via transgenic approach and marker assisted selection. PMID- 21621743 TI - Introgression of qPE9-1 allele, conferring the panicle erectness, leads to the decrease of grain yield per plant in japonica rice (Oryza sativa L.). AB - Panicle architecture is closely related to yield formation. The qPE9-1 gene has been proved to be widely used in high-yield rice cultivar developments, conferring erect panicle character in japonica rice. Recently, qPE9-1 has been successfully cloned; however, the genetic effect on grain yield per plant of the erect panicle allele qPE9-1 is controversial yet. In the present study, a drooping panicle parent Nongken 57, carrying qpe9-1 allele, was used as recurrent parent to successively backcross to a typical erect panicle line from the double haploid (DH) population (Wuyunjing 8/Nongken 57), which was previously shown to carry qPE9-1 allele. Thus a pair of near-isogenic lines (NILs) was developed. The comparison of agronomic traits between the NILs showed that, when qpe9-1 was replaced by qPE9-1, the panicle architecture was changed from drooping to erect; moreover, the panicle length, plant height, 1000-grain weight and the tillers were significantly decreased, consequently resulting in the dramatic decrease of grain yield per plant by 30%. Therefore, we concluded that the qPE9-1 was a major factor controlling panicle architecture, and qPE9-1 had pleiotropic nature, with negative effects on grain yield per plant. This result strongly suggests that the erect panicle allele qPE9-1 should be used together with other favorable genes in the high-yield breeding practice. In addition, the effect of qPE9-1 on eating and cooking quality was also discussed in the present study. PMID- 21621744 TI - Fusional convergence in childhood intermittent exotropia. AB - PURPOSE: To evaluate fusional convergence and associations with control in children with intermittent exotropia (XT). DESIGN: Retrospective case series. METHODS: Sixty-four children (median age 7 years) with intermittent XT were identified with measures of angle of deviation, control (using a previously reported 0-to-5-point scale), and fusional convergence at a single examination. Total convergence was defined as the sum of the angle of deviation plus additional convergence in reserve. Mean values were compared with published normals. A fusion reserve ratio was calculated as "convergence reserve divided by angle of deviation." Relationships of control score with total amplitude, reserve, recovery, and fusion reserve ratio were assessed using Spearman rank correlations. RESULTS: Convergence differed from normals in children with intermittent XT: total convergence was higher at distance (33 prism diopters [pd] vs 17 pd, P<.0001) and near (38 pd vs 18 pd, P<.0001) whereas convergence reserve was lower at distance (7 pd vs 17 pd; P<.0001). There was a strong correlation between fusion reserve ratio and control score at distance (R=-0.75, P<.0001) and near (R=-0.66, P<.0001). CONCLUSIONS: Children with intermittent XT have subnormal convergence reserves at distance. The fusion reserve ratio correlates well with control and may be useful in grading the severity of intermittent XT. PMID- 21621745 TI - The effects of user factors and symbol referents on public symbol design using the stereotype production method. AB - This study investigated the influence of user factors and symbol referents on public symbol design among older people, using the stereotype production method for collecting user ideas during the symbol design process. Thirty-one older adults were asked to draw images based on 28 public symbol referents and to indicate their familiarity with and ease with which they visualised each referent. Differences were found between the pictorial solutions generated by males and females. However, symbol design was not influenced by participants' education level, vividness of visual imagery, object imagery preference or spatial imagery preference. Both familiar and unfamiliar referents were illustrated pictorially without much difficulty by users. The more visual the referent, the less difficulty the users had in illustrating it. The findings of this study should aid the optimisation of the stereotype production method for user-involved symbol design. PMID- 21621746 TI - Interpretation bias in Cluster-C and borderline personality disorders. AB - Cognitive therapy (CT) assumes that personality disorders (PDs) are characterized by interpretational biases that maintain the disorder. Changing interpretations is therefore a major aim of CT of PDs. This study tested whether Borderline PD (BPD), Avoidant and Dependent PD (AV/DEPD), and Obsessive-Compulsive PD (OCPD) are characterized by specific interpretations. Among the 122 participants there were 55 PD patients (17 BPD, 30 AV/DEPD, 29 OCPD diagnoses), 26 axis-1 patients, and 41 nonpatients. Participants put themselves into 10 scripts of negative events and noted feelings, thoughts and behaviors that came to mind. Next, they chose between hypothesized BPD-specific, AV/DEPD-specific, and OCPD-specific interpretations of each event (forced choice). Lastly, participants rated belief in each interpretation. Regression analyses revealed that forced choices and belief ratings supported the CT-model of BPD and AV/DEP: interpretations were specific. The alleged OCPD-beliefs were however not specifically related to OCPD, with relatively high popularity in axis-1 patients and nonpatients. The open responses were classified by judges blind for diagnoses, with the following results. BPD was characterized by low levels of solution-focused and healthy flexible/accepting responses, and higher levels of criticizing others and malevolent interpretations of others. AV/DEPD was characterized by lower levels of solution-focused responses, and higher levels of self-criticism, negative emotions, guilt and fear of judgment, as well as lower levels of other-criticism. OCPD only showed trends for lower healthy responses, and higher compulsiveness and worry. It is concluded that the assumptions of CT are supported for BPD and AV/DEPD, but not - at least not on the explicit interpretational level - for OCPD. CT of OCPD might need a slightly different approach. PMID- 21621747 TI - Degree and pattern of calbindin immunoreactivity in granule cells of the dentate gyrus differ in mesial temporal sclerosis, cortical malformation- and tumor related epilepsies. AB - A loss of calbindin immunoreactivity in granule cells of the hippocampal dentate gyrus is a characteristic feature of temporal lobe epilepsy with hippocampal sclerosis. Whether decreased calbindin expression is unique to the hippocampal sclerosis associated with cryptogenic temporal lobe epilepsy, or also occurs in tumor- or malformation-related epilepsy, is unknown. We show that calbindin immunoreactivity in granule cells has been decreased in epilepsy regardless of its etiology. In cases of cortical malformations or hippocampal sclerosis, calbindin immunoreactivity was undetectable in most granule cells. In tumor related resections, in patients who had a long history of epileptic seizures, calbindin was detected only in one-third of granule cells. Regardless of etiology, calbindin expression correlated with age of onset and with duration of the epilepsy. In contrast to tumor-induced epilepsy, where calbindin immunoreactive granule cells were equally distributed in the granule cell layer, in hippocampal sclerosis and malformation-related epilepsy, two-thirds of calbindin-immunoreactive granule cells were located in the outer half and only one-third in the inner half of the layer. Developmentally, granule cells at the border of the molecular layer are ontogenetically the oldest, and those at the border of the hilus are the youngest. The reduction of calbindin immunoreactivity in ontogenetically younger granule cells highlights the deleterious effect of early occurring epilepsy and initial early precipitating injury, including febrile seizures that may substantially affect developing immature granule cells, but less the earlier born matured ones. PMID- 21621748 TI - Calcium cooperativity of exocytosis as a measure of Ca2+ channel domain overlap. AB - The number of Ca(2+) channels contributing to the exocytosis of a single neurotransmitter vesicle in a presynaptic terminal has been a question of significant interest and debate, and is important for a full understanding of localized Ca(2+) signaling in general, and synaptic physiology in particular. This is usually estimated by measuring the sensitivity of the neurotransmitter release rate to changes in the synaptic Ca(2+) current, which is varied using appropriate voltage-clamp protocols or via pharmacological Ca(2+) channel block under the condition of constant single-channel Ca(2+) current. The slope of the resulting log-log plot of transmitter release rate versus presynaptic Ca(2+) current is termed Ca(2+)current cooperativity of exocytosis, and provides indirect information about the underlying presynaptic morphology. In this review, we discuss the relationship between the Ca(2+) current cooperativity and the average number of Ca(2+) channels participating in the exocytosis of a single vesicle, termed the Ca(2+)channel cooperativity. We relate these quantities to the morphology of the presynaptic active zone. We also review experimental studies of Ca(2+) current cooperativity and its modulation during development in different classes of synapses. PMID- 21621749 TI - Scar ablation combined with LP/OEC transplantation promotes anatomical recovery and P0-positive myelination in chronically contused spinal cord of rats. AB - We have successfully removed an existing glial scar in chronically contused rat spinal cord using a rose Bengal-based phototoxic method. The purpose of this study is to examine if scar ablation benefits the anatomical recovery by cell/tissue transplantation, and thus provides a more permissive physical and biochemical environment for axonal growth, which may lead to functional recovery. Immediately after scar ablation, we transplanted lamina propria (LP) of the olfactory mucosa alone or in combination with cultured olfactory ensheathing cells (OEC) into the lesion cavity 6 weeks after contusion injury (NYU impactor device, 25 mm height setting) at spinal cord segment T10 of adult female Long Evans rats. Sixteen weeks after scar ablation and transplantation, we found that the initial repaired tissue significantly expanded, companied by remarkable reduction or disappearance of the lesion cavity and integration of repaired tissue with the spared tissue, thus resulting in histological repair of damaged cord tissue at the injury epicenter. Glial scar reformation was effectively prevented after ablation due to the tissue repair. In addition, at the injury epicenter P0 (myelin glycoprotein P-zero)-positive myelination formed by Schwann cells, which are known to myelinate regenerating and demyelinated axons, were significantly increased in number compared with the control animals. However, when evaluated with BBB open-field scale a significant improvement of locomotor function was not observed in this study; the possible reasons were discussed. PMID- 21621750 TI - Synthesis of blockwise alkylated (1->4) linked trisaccharides as surfactants: influence of configuration of anomeric position on their surface activities. AB - New carbohydrate-based surfactants consisting of hydrophilic cellobiosyl and hydrophobic glucosyl residues, methyl beta-d-glucopyranosyl-(1->4)-alpha-d glucopyranosyl-(1->4)-2,3,6-tri-O-methyl-alpha-d-glucopyranoside 1 (GbetaGalphaMalpha, G: glucopyranosyl residue, alpha and beta: alpha-(1->4)- and beta-(1->4) glycosidic bonds, M: methyl group), 2 (G(beta)G(beta)M(alpha)), 3 (G(beta)G(alpha)M(beta)), 4 (G(beta)G(beta)M(beta)), 5 (G(beta)G(alpha)E(alpha), E: ethyl group), 6 (G(beta)G(beta)E(alpha)), 7 (G(beta)G(alpha)E(beta)), 8 (G(beta)G(beta)E(beta)) and eight alpha-and beta-glycoside mixtures (a mixture of 1 and 2: 1/2=62/38 (9), 32/68 (10); a mixture of 3 and 4: 3/4=69/31 (11), 32/68 (12); a mixture of 5 and 6: 5/6=62/38 (13), 33/67 (14); a mixture of 7 and 8: 7/8=59/41 (15), 29/71 (16)) were synthesized via combined methods consisting of acid-catalyzed alcoholysis of cellulose ethers and glycosylation of phenyl thio cellobioside derivatives. Their surface activities in aqueous solution depended on their chemical structures: alpha- or beta-(1->4) linkage between hydrophilic cellobiosyl and hydrophobic glucosyl blocks, methyl or ethyl groups of hydrophobic glucosyl block, and alpha- or beta-linked ether group at the C-1 of hydrophobic glucosyl block. The mixing effect of alpha- and beta-glycosides on surface activities was also investigated. As a result, ethyl beta-d glucopyranosyl-(1->4)-alpha-d-glucopyranosyl-(1->4)-2,3,6-tri-O-ethyl-beta-d glucopyranoside 7 (G(beta)G(alpha)E(beta)) had the highest surface activity, and its critical micellar concentration (CMC) and gamma(CMC) (surface tension at CMC) values of compound 7 were 0.5mM (ca. 0.03wt%) and 34.5mN/m, respectively. The surface tensions of alpha- and beta-glycoside mixtures except for compounds 9 and 10 were almost equal to those of pure compounds. The syntheses of the mixtures of alpha- and beta-glycosides without purification process are easier than those of pure compounds. Thus, the mixtures should be more practical compounds for industrial use as a surfactant. PMID- 21621751 TI - A novel DTPA cross-linking of hyaluronic acid and metal complexation thereof. AB - Macromolecular conjugates of a natural polysaccharide, hyaluronic acid, with diethylenetriaminepentaacetic acid (DTPA)-metal complexes were synthesized and characterized by FTIR, NMR, SEC-MALLS and ICP analysis. Several parameters of the cross-linking reaction as molecular weight of starting HA, temperature, equivalent of DTPA bis-anhydride, concentration of HA, presence of transacylation catalyst DMAP and reaction time were studied. The mechanism for the reaction was suggested and relationship between the molecular weight assigned by SEC-MALLS, reaction parameters and rheological properties of the final cross-linked products were investigated. PMID- 21621752 TI - Complete (1)H and (13)C NMR chemical shift assignments of mono-, di-, and trisaccharides as basis for NMR chemical shift predictions of polysaccharides using the computer program casper. AB - The computer program casper uses (1)H and (13)C NMR chemical shift data of mono- to trisaccharides for the prediction of chemical shifts of oligo- and polysaccharides. In order to improve the quality of these predictions the (1)H and (13)C, as well as (31)P when applicable, NMR chemical shifts of 30 mono-, di , and trisaccharides were assigned. The reducing sugars gave two distinct sets of NMR resonances due to the alpha- and beta-anomeric forms. In total 35 (1)H and (13)C NMR chemical shift data sets were obtained from the oligosaccharides. One- and two-dimensional NMR experiments were used for the chemical shift assignments and special techniques were employed in some cases such as 2D (1)H,(13)C-HSQC Hadamard Transform methodology which was acquired approximately 45 times faster than a regular t(1) incremented (1)H,(13)C-HSQC experiment and a 1D (1)H,(1)H CSSF-TOCSY experiment which was able to distinguish spin-systems in which the target protons were only 3.3Hz apart. The (1)H NMR chemical shifts were subsequently refined using total line-shape analysis with the PERCH NMR software. The acquired NMR data were then utilized in the casper program (http://www.casper.organ.su.se/casper/) for NMR chemical shift predictions of the O-antigen polysaccharides from Klebsiella O5, Shigella flexneri serotype X, and Salmonella arizonae O62. The data were compared to experimental data of the polysaccharides from the two former strains and the lipopolysaccharide of the latter strain showing excellent agreement between predicted and experimental (1)H and (13)C NMR chemical shifts. PMID- 21621753 TI - Observing shared attention modulates gaze following. AB - Humans' tendency to follow others' gaze is considered to be rather resistant to top-down influences. However, recent evidence indicates that gaze following depends on prior eye contact with the observed agent. Does observing two people engaging in eye contact also modulate gaze following? Participants observed two faces looking at each other or away from each other before jointly shifting gaze to one of two locations. Targets appeared either at the cued location or at the non-cued location. In three experiments gaze cueing effects (faster responses to objects appearing at the cued location) were found only when the two faces had looked at each other before shifting gaze. In contrast, no effects of gaze following were observed when the two faces had looked away from each other. Thus, the attentional relation between observed people modulates whether their gaze is followed. PMID- 21621754 TI - Prevalence of depression and its relationship with other clinical characteristics in a sample of patients with stable schizophrenia. AB - AIM: To evaluate the prevalence of depression using the Calgary Depression Scale for Schizophrenia (CDSS) in a sample of Spanish patients with stable schizophrenia and without a diagnosis of depression. METHODS: We included stable outpatients of 18 to 50 years of age, with a diagnosis of schizophrenia, schizoaffective disorder, or schizophreniform disorder who had not been diagnosed with depression. In this cross-sectional study, we administered the CDSS, the Positive and Negative Syndrome Scale (PANSS), the Scale for the Assessment of Negative Symptoms (SANS), the Scale to Assess Unawareness of Mental Disorder (SUMD), the Simpson Angus Scale (SAS), and the Barnes Akathisia Rating Scale (BARS). RESULTS: A total of 95 patients were recruited, 90 of them were included in the statistical analysis. Twenty-eight patients had a total score of 5 or more points on the CDSS, making the prevalence of depression 31% (95% confidence interval, 22-41). The CDSS had a high correlation with the depressive factor of the PANSS and a moderate correlation with the general psychopathology subscale of the PANSS. The correlation of the CDSS total score with negative symptoms was moderate using the SANS and low with the PANSS-negative. There was no correlation between depressive symptoms and positive symptoms, insight, and extrapyramidal symptoms; and the correlation with akathisia was low. CONCLUSION: Our results suggest that patients with stable schizophrenia who have not been diagnosed with depression frequently have clinically relevant symptoms of depression, and that these symptoms, with the possible exception of a contribution from negative symptoms, are not secondary to other symptoms of their disorder or to extrapyramidal adverse effects of medications. PMID- 21621755 TI - Factor structure of the Center for Epidemiologic Studies Depression Scale in Taiwanese adolescents. AB - BACKGROUND: The original study of Radloff (Appl Psychol Meas. 1977. 1:385-401) on the Center for Epidemiologic Studies Depression Scale (CES-D) indicated a 4 factor model in the adult population. However, the factor structure of the CES-D in Asian adolescents has not been extensively validated. The aim of this study was to examine the factor structure of the Chinese version of the CES-D in a large representative Taiwanese adolescent population. METHOD: A total of 10 116 adolescents completed the Chinese version of the CES-D. We used confirmatory factor analysis to examine the adequacy of 5 models of the factor structure in adolescents who were divided into 4 groups by sex and age. We also enrolled the variables of suicide tendency, insomnia, and peer relationships into the confirmatory factor analysis to examine the factor structure of the CES-D and examined the correlations between the CES-D factors and these variables. RESULTS: The results of this study indicated that the four-factor model (depressed affect, somatic symptoms, interpersonal problems, and positive affect) had the highest validity in Taiwanese adolescents. We also found that although the 4 factors of the CES-D were correlated with each other, their correlations with suicide tendency, insomnia, and peer relationships were different. CONCLUSIONS: This study supported the usefulness of the Chinese version of the CES-D as a tool to understand the concept of depression in Taiwanese adolescents. PMID- 21621756 TI - Model-based assessment of baroreflex and cardiopulmonary couplings during graded head-up tilt. AB - We propose a multivariate dynamical adjustment (MDA) modeling approach to assess the strength of baroreflex and cardiopulmonary couplings from spontaneous cardiovascular variabilities. Open loop MDA (OLMDA) and closed loop MDA (CLMDA) models were compared. The coupling strength was assessed during progressive sympathetic activation induced by graded head-up tilt. Both OLMDA and CLMDA models suggested that baroreflex coupling progressively increased with tilt table inclination. Only CLMDA model indicated that cardiopulmonary coupling due to the direct link from respiration to heart period gradually decreased with tilt table angles, while that due to the indirect link mediated by systolic arterial pressure progressively increased. PMID- 21621757 TI - Computer simulations on multiprobe freezing of irregularly shaped tumors. AB - Cryosurgery is particularly suitable for the treatment of unresectable liver tumors. However, a major bottleneck is encountered during the treatment of large sized irregularly shaped tumors. Large and complex liver tumors have varying degree of shape irregularity. Adopting a multiprobe freezing model, simulations for an irregularly shaped liver tumor were conducted. The model, validated with both in-vitro data from an experimental setup, showed good agreement of up to 5.8%. The chosen mathematical treatment and simulation technique permit the study of employing multiple cryoprobes to destroy cancer cells in irregularly shaped tumors. Results from our study indicated that multiple cryoprobes can be strategically positioned to form ice-fronts with various contours that adhere to the shape and size of the tumor. The amount of cell-deaths within the tumor after the -50 degrees C ice-front can be quantitatively calculated to determine the efficacy of different multiprobe arrangements in order to maximize cell destruction. The paper also underlines a piecewise approach of using several cryoprobes to quickly 'sculpt' the desired shape of the ice-front based on the physical morphology of an irregular-shaped tumor. This numerical study forms an essential framework in allowing surgeons to make informed decisions on the most effective surgical protocol based on the degree of irregularity in shape and size of tumor. PMID- 21621758 TI - Theoretical study of 3-D molecular similarity and ligand binding modes of orthologous human and rat D2 dopamine receptors. AB - The D(2) dopamine receptor (D(2)DR) is an important target for the treatment of some central nervous system disorders, such as Parkinson disease, schizophrenia and drug-dependence. In this work, we built 3-D models of the long form of human and rat D(2)DRs by considering data from the crystallized D3 dopamine receptor, beta2 adrenoceptor and A2a adenosine receptor as templates. Then, docking was performed with ligand and protein residue flexibility. These results were used to analyze ligand recognition and estimate binding affinity. Our results show that the predicted ligand affinity correlates with experimental data, and binding modes are very similar between the D(2)DRs of these two species. PMID- 21621759 TI - Automatic identification of fetal breathing movements in fetal RR interval time series. AB - Fetal breathing movements are associated with respiratory sinus arrhythmia (RSA). We present an algorithm which processes RR interval time series in the time and frequency domain, identifying spectral peaks with characteristics consistent with fetal RSA. Tested on 50 data sets from the second and third trimester, the algorithm had a sensitivity of 96.1%, false positive rate 35.7%, false negative rate 3.9%. The characteristics of automatically and visually identified episodes were very similar and corresponded the expected changes over gestation. The method is suited for easy and reliable identification of fetal breathing movements. PMID- 21621760 TI - Neurocognitive disorder detection based on feature vectors extracted from VBM analysis of structural MRI. AB - Dementia is a growing concern due to the aging process of the western societies. Non-invasive detection is therefore a high priority research endeavor. In this paper we report results of classification systems applied to the feature vectors obtained by a feature extraction method computed on structural magnetic resonance imaging (sMRI) volumes for the detection of two neurological disorders with cognitive impairment: myotonic dystrophy of type 1 (MD1) and Alzheimer disease (AD). The feature extraction process is based on the voxel clusters detected by voxel-based morphometry (VBM) analysis of sMRI upon a set of patient and control subjects. This feature extraction process is specific for each kind of disease and is grounded on the findings obtained by medical experts. The 10-fold cross validation results of several statistical and neural network based classification algorithms trained and tested on these features show high specificity and moderate sensitivity of the classifiers, suggesting that the approach is better suited for rejecting than for detecting early stages of the diseases. PMID- 21621761 TI - Blood oxygenation changes resulting from trains of low frequency transcranial magnetic stimulation. AB - The evoked responses to transcranial magnetic stimulation (TMS) have been previously demonstrated to be on average greater at the beginning of a session; however the physiological reason for this remains uncertain. In order to investigate a possible hemodynamic mechanism for this phenomenon, changes in oxy hemoglobin (HbO) following trains of single pulse TMS was investigated using near infra-red spectroscopy (NIRS). TMS was delivered in trains of two and four pulses to left pre-frontal cortex (PFC) at a typical intensity and frequency (.2 Hz) used in neuroscience research. Both trains resulted in significant drops of HbO that remained after the cessation of TMS. The changes observed imply that arterial supply drops following suprathreshold TMS and oxygen consumption outstrips supply, resulting in a net drop of HbO. This study provides evidence that at typical TMS delivery frequencies, local HbO levels remain at a sustained lower level than at the beginning of the session, potentially explaining changes in sensitivity to stimulation with repeated TMS pulses. PMID- 21621762 TI - The "frontal syndrome" revisited: lessons from electrostimulation mapping studies. AB - For a long time, in a localizationist view of brain functioning, a combination of symptoms called "frontal syndrome" has been interpreted as the direct result of damages involving the frontal lobe(s). The goal of this review is to challenge this view, that is, to move to a hodotopical approach to lesion mapping, on the basis of new insights provided by intraoperative electrostimulation mapping investigations in patients who underwent awake surgery for cerebral tumors. These original data reported in the last decade break with the traditional dogma of a modular and fixed organization of the central nervous system, by switching to the concepts of cerebral connectivity and plasticity - i.e., a brain organization based on dynamic interrelationships between parallel distributed networks. According to this revisited model, "frontal symptoms" can be generated by tumor or electrostimulation not only of the frontal lobes, but also of cortical and subcortical (white matter pathways/deep gray nuclei) structures outside the frontal lobes: especially, stimulation of the superior longitudinal fascicle may elicit speech production disorders, syntactic disturbances, involuntary language switching or phonemic paraphasia (arcuate fascicle), stimulation of the inferior fronto-occipital fascicle can generate semantic paraphasia or deficit of cross modal judgment, stimulation of the subcallosal fasciculus may elicit transcortical motor aphasia, while stimulation of the striatum induces preservations. On the other hand, it is also possible to perform extensive right or left frontal lobectomy in patients who continue to have a normal familial, social and professional life, without "frontal syndrome". Therefore, this provocative approach may open the door to a renewal in the modeling of brain processing as well as in its clinical applications, especially in the fields of cerebral surgery and functional rehabilitation. These findings illustrate well the need to reinforce links between cognitive neuroscience and clinical neurology/neurosurgery. PMID- 21621763 TI - Neuroimaging--chance effects and unintended consequences. PMID- 21621764 TI - Conceptual representations in mind and brain: theoretical developments, current evidence and future directions. AB - Conceptual representations in long-term memory crucially contribute to perception and action, language and thought. However, the precise nature of these conceptual memory traces is discussed controversially. In particular, the grounding of concepts in the sensory and motor brain systems is the focus of a current debate. Here, we review theoretical accounts of the structure and neural basis of conceptual memory and evaluate them in light of recent empirical evidence. Models of conceptual processing can be distinguished along four dimensions: (i) amodal versus modality-specific, (ii) localist versus distributed, (iii) innate versus experience-dependent, and (iv) stable versus flexible. A systematic review of behavioral and neuroimaging studies in healthy participants along with brain damaged patients will then be used to evaluate the competing theoretical approaches to conceptual representations. These findings indicate that concepts are flexible, distributed representations comprised of modality-specific conceptual features. Conceptual features are stored in distinct sensory and motor brain areas depending on specific sensory and motor experiences during concept acquisition. Three important controversial issues are highlighted, which require further clarification in future research: the existence of an amodal conceptual representation in the anterior temporal lobe, the causal role of sensory and motor activation for conceptual processing and the grounding of abstract concepts in perception and action. We argue that an embodiment view of conceptual representations realized as distributed sensory and motor cell assemblies that are complemented by supramodal integration brain circuits may serve as a theoretical framework to guide future research on concrete and abstract concepts. PMID- 21621765 TI - Accuracy, reliability, validity and limitations of functional and structural magnetic resonance imaging data. PMID- 21621766 TI - New fast mismatch negativity paradigm for determining the neural prerequisites for musical ability. AB - Studies have consistently shown that the mismatch negativity (MMN) for different auditory features correlates with musical skills, and that this effect is more pronounced for stimuli integrated in complex musical contexts. Hence, the MMN can potentially be used for determining the development of auditory skills and musical expertise. MMN paradigms, however, are typically very long in duration, and far from sounding musical. Therefore, we developed a novel multi-feature MMN paradigm with 6 different deviant types integrated in a complex musical context of no more than 20 min in duration. We found significant MMNs for all 6 deviant types. Hence, this short objective measure can putatively be used as an index for auditory and musical development. PMID- 21621767 TI - A family with Xq22.3q25 interstitial deletion and normal ovarian function. AB - OBJECTIVE: To investigate genomic changes in a family with deletion of X chromosome q22.3-q25 associated with normal constitutional and reproductive phenotypes. DESIGN: Case report. SETTING: Academic district hospital genetic laboratory. PATIENT(S): A family incidentally found to have deletion of X chromosome q22.3-q25. INTERVENTION(S): Cytogenetic analysis and array-based comparative genomic hybridization for amniotic fluid and peripheral blood lymphocyte of family members. MAIN OUTCOME MEASURE(S): Ovarian function and menstrual cycles. RESULT(S): The proband and two daughters showed deletion of Xq22.3q25. This region spans 17.4 Mb and contains 121 genes. CONCLUSION(S): Female subjects with deletion of Xq22.3q25 may present with normal constitutional and reproductive phenotypes. PMID- 21621768 TI - Pregnancy outcomes by intravaginal and intrauterine insemination in 82 couples with male factor infertility due to spinal cord injuries. AB - OBJECTIVE: To report our center's pregnancy rates (PR) by intravaginal insemination (IVI) or intrauterine insemination (IUI) in 82 couples with male partners with spinal cord injuries. DESIGN: Retrospective analysis. SETTING: Major medical center. PATIENT(S): Male patients with spinal cord injuries and their female partners. INTERVENTION(S): Intravaginal insemination and IUI. MAIN OUTCOME MEASURE(S): Pregnancy and live birth outcomes. RESULT(S): Overall, 31 of the 82 couples (37.8% PR) achieved 39 pregnancies. Sperm were obtained by masturbation, penile vibratory stimulation, or electroejaculation in 4 men (4.9%), 42 men (51.2%), and 36 men (43.9%), respectively. Intravaginal insemination, performed mostly at home by selected couples, was undertaken in 45 couples, 17 of whom (37.8% PR) achieved 20 pregnancies. Intrauterine insemination was performed in 57 couples, 14 of whom (24.6% PR) achieved 19 pregnancies, with a cycle fecundity of 7.9%. Eighteen and 21 live births occurred by IVI and IUI, respectively. CONCLUSION(S): The methods of IVI and IUI are reasonable options for this patient population. These methods warrant consideration before proceeding to assisted reproductive technologies (ART). PMID- 21621769 TI - The WHO, WHY, WHAT, WHEN, WHERE, and HOW of clinical trial registries. AB - Clinical trials must be appropriately registered prior to patient enrollment to be considered for publication by most top journals, including Fertility and Sterility. Clinical trials involving FDA regulated drugs, whether investigational or already approved, must be registered by law, and summary results must be posted in the registry whether ultimately published or not. This review provides detailed information for authors regarding these requirements and a discussion as to their importance for acceptance of a manuscript and for reviewers to be assured of the integrity of the trial design, execution, and analysis. The accompanying review on study design and statistics is meant to illustrate the difficulties reviewers experience in evaluating a manuscript that does not follow these requirements. The editors of Fertility and Sterility reiterate that clinical trials that have not been registered will not be accepted for review. PMID- 21621770 TI - Cumulative live-birth rates per total number of embryos needed to reach newborn in consecutive in vitro fertilization (IVF) cycles: a new approach to measuring the likelihood of IVF success. AB - OBJECTIVE: To report the use of cumulative live-birth rates (CLBRs) per ovarian stimulation cycle to measure the success of IVF is proving to be the most accurate method for advising couples who failed to conceive, although the accuracy yielded is relatively low, and cycle outcome is highly dependent on the number of embryos replaced. Our aim with this work is to report the CLBRs of IVF as a function of the number of embryos required to reach a live birth (EmbR), considering age, day of ET, and infertility etiology. DESIGN: Survival curves and Kaplan-Meier methods to analyze CLBR in a retrospective cohort with respect to the number of EmbR. SETTING: University-affiliated infertility center. PATIENT(S): Infertile couples undergoing IVF using own oocytes. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): CLBR per embryo transferred. RESULT(S): CLBRs increase rapidly between 1 and 5 EmbR, moderately between 5 and 15, and slowly thereafter. Live-birth rates rise more slowly when embryos are transferred on days 2-3 rather than on days 5-6, with comparable long-term results. Women's age is a negative factor from 35 to 37 years old, with a dramatic decrease in live birth rates beyond age 40 years. In addition, there are significant worse results in endometriosis patients. CONCLUSION(S): The relationship between CLBR and number of EmbR provides realistic and precise information regarding IVF success and can be used to guide couples and practitioners. PMID- 21621772 TI - Intravenous calcium infusion as a novel preventive therapy of ovarian hyperstimulation syndrome for patients with polycystic ovarian syndrome. AB - OBJECTIVE: To evaluate the effectiveness of IV calcium infusion on prevention of ovarian hyperstimulation syndrome (OHSS) in patients with polycystic ovary syndrome undergoing assisted reproductive techniques cycles. DESIGN: A retrospective comparative study. SETTING: Assisted reproduction techniques centre in Turkey. PATIENT(S): Four hundred fifty-five women with high risk for OHSS. INTERVENTION(S): The patients in group I (n = 84) were administered IV calcium gluconate for prevention of OHSS, and the patients in group II (n = 371) comprised the control group, with no manipulation for prevention of OHSS and were age- and body mass index-matched with the study group. MAIN OUTCOME MEASURE(S): Ovarian hyperstimulation syndrome rate, clinical pregnancy rate. RESULT(S): Mean (+/-SD) ages of the women in the calcium infusion group (group I) and the control group (group II) were comparable (30.5 +/- 4.3 vs. 31.4 +/- 3.9, respectively). Ovarian hyperstimulation syndrome was found in 16.2% (60 patients) in group II, whereas in group I, only 3 patients (3.6%) developed OHSS. Interestingly, all the hyperstimulation cases in group I were mild, and there was no severe effect. Implantation rates were similar in both groups. Furthermore, we obtained clinical pregnancy in nearly 40.5% in group I and 28.8% in group II. The live-birth rate was 38.1% in the calcium infusion group and 24.8% in the control group. CONCLUSION(S): Intravenous calcium infusion resulted in a significantly lower rate of development of OHSS for patients with polycystic ovary syndrome and high risk of OHSS. This novel therapy may be used for prevention of OHSS effectively. PMID- 21621771 TI - Quantification of the impact of endometriosis symptoms on health-related quality of life and work productivity. AB - OBJECTIVE: To quantify the impact of endometriosis-related symptoms on physical and mental health status, health-related quality of life, and work-related aspects (absenteeism, presenteeism, work productivity, and activity impairment). DESIGN: Cross-sectional quantitative study. SETTING: Academic and research institution. PATIENT(S): Women (n = 193) with self-reported surgically diagnosed endometriosis from the Endometriosis Patient Registry at Ponce School of Medicine and Health Sciences (PSMHS). INTERVENTION(S): Anonymous questionnaire divided into three sections consisting of questions from the Patient Health Survey (SF 12), the Endometriosis Health Profile (EHP-5), and the Work Productivity and Activity Impairment Survey (WPAI). MAIN OUTCOME MEASURE(S): Quantification of impact of endometriosis symptoms on physical and mental health status, health related quality of life, absenteeism, presenteeism, work productivity, and activity impairment. RESULT(S): Patients had SF-12 scores denoting statistically significant disability in the physical and mental health components. They also reported an average of 7.41 hours (approximately one working day) of work time lost during the week when the symptoms are worse. In addition, the WPAI scores showed a high impact on work-related domains: 13% of average loss in work time (absenteeism), 65% of work impaired (presenteeism), 64% of loss in efficiency levels (work productivity loss), and 60% of daily activities perturbed (activity impairment). CONCLUSION(S): Endometriosis symptoms such as chronic, incapacitating pelvic pain and infertility negatively and substantially impact the physical and mental health status, health-related quality of life, and productivity at work of women. PMID- 21621773 TI - Ultrasound ovarian assessments after endometrioma ablation using plasma energy. AB - We performed a retrospective three-dimensional ultrasound evaluation of the ovarian features in ten women with no previous ovarian surgery who benefited from ablation by plasma energy for unilateral endometriomas greater than to 30 mm in diameter. Values of ovarian volume and antral follicle count in operated ovaries were decreased by an average 12% and 18%, respectively, suggesting that endometrioma ablation using plasma energy spares the underlying ovarian parenchyma. PMID- 21621774 TI - The association between semen quality in workers and the concentration of di(2 ethylhexyl) phthalate in polyvinyl chloride pellet plant air. AB - OBJECTIVE: To investigate potential associations between semen quality in workers and the concentrations of di(2-ethylhexyl) phthalate (DEHP) in personal air collected from polyvinyl chloride (PVC) plants. DESIGN: Cross-sectional study. SETTING: PVC plants in Taiwan. PATIENT(S): Forty-five male workers employed in two PVC pellet plants. INTERVENTION(S): None. MAIN OUTCOME MEASUREMENT(S): Sperm concentration, motility, morphology, and chromatin DNA integrity were accessed. RESULT(S): The workers were divided into low- and high-DEHP-exposed groups in accordance with the median levels of DEHP (23.7 MUg/m(3)) in personal air. In the high-DEHP-exposed group, significant increases were found in the tendency for sperm DNA denaturation (alphaT) induction, the DNA fragmentation index (DFI), and propensity for coffee drinking. After adjusting for coffee drinking, cigarette smoking, and age, personal air concentrations of DEHP showed positive associations with alphaT (beta = 0.038) and DFI (beta = 0.140) and negative associations with sperm motility (beta = -0.227). CONCLUSION(S): This is the first study to demonstrate a link between DEHP concentration in ambient air and the adverse effects in sperm motility and chromatin DNA integrity. Given the current wide use of these PVC products, the implications for phthalates toxicity and occupational health could be considerable. PMID- 21621775 TI - Predictors of home death of home palliative cancer care patients: a cross sectional nationwide survey. AB - OBJECTIVES: To identify factors influencing the place of death among home palliative cancer care patients, focusing on the role of nurses in terms of pre- and post-discharge from hospital to home care settings. DESIGN, SETTINGS AND PARTICIPANTS: A cross-sectional nationwide questionnaire survey was conducted at 1000 randomly selected homecare agencies in Japan. The questionnaires were completed by primary community nurses of home palliative patients just after their discharge. A total of 568 responses were analyzed (effective response rate, 69%). RESULTS: Multivariate logistic regression analysis revealed the following independent factors of place of death among those patients: desire for home death at referral by both patient and family caregiver; caregiver relationship to patient as daughter or daughter-in-law; totally bedridden functional status of patient; patient not suffering from depression and/or anxiety at referral; patients and caregivers duly informed about the dying process/death in detail, as well as instructed by community nurses about pain management and how to treat/prevent bedsores in home care settings. CONCLUSIONS: This study demonstrated the importance of both the hospital and community nurses' role in increasing the patients' chance of dying at home. Hospital nurses should support early transfer to home palliative care according to their assessment of the desire of patient/family caregiver for home death, the patients' clinical status, and caregivers' ability to provide patient care at home. Community nurses should inform patients/family caregiver in detail about the dying process/death just after discharge, relieve patient pain, treat/prevent bedsores, and instruct family caregivers on their symptom control. PMID- 21621776 TI - Interaction of the inflammasome genes CARD8 and NLRP3 in abdominal aortic aneurysms. AB - OBJECTIVE: Cholesterol crystals have been shown to cause inflammation, and ultimately atherosclerotic lesions through the activation of the NLRP3 inflammasome. As cholesterol crystals have also been found in the walls of patients with abdominal aortic aneurysms (AAA), it is possible that the NLRP3 inflammasome is involved in AAA and genetic variability within this protein complex could alter disease risk. The primary objective of this study was to assess whether there is genetic evidence for a role of the NLRP3 inflammasome in AAA by testing for association of AAA with functional single nucleotide polymorphisms (SNPs) in the CARD8 and NLRP3 genes. METHODS: AAA patients (n=1151) and controls (n=727) were genotyped for CARD8 SNP rs2043211 and NLRP3 SNP rs35829419 using TaqMan SNP assays. IL1-beta, C-reactive protein (CRP), and lipoprotein (a) [Lp(a)] were measured in the plasma of a subset of study participants. The Kruskal-Wallis Rank test was conducted to test for differences in mean concentration of IL1-beta, CRP and Lp(a). Logistic regression was used to test for interaction between CARD8 and NLRP3. RESULTS: Significantly higher mean concentration of plasma IL1-beta was observed in study participants who were homozygous for the common C allele of NLRP3 rs35829419 (p=0.010). Interaction between rs2043211 and rs35829419 was observed in this dataset (chi(2)=6.22; p=0.044), which strengthened when adjusted for age, gender, smoking, diabetes, hypertension, and dyslipidemia (chi(2)=14.75; p=0.012); and separately for NOD2 genotype (chi(2)=14.06; p=0.015). CONCLUSION: Our finding suggests genetic variability within the NLRP3 inflammasome may be important in the pathophysiology of AAA. PMID- 21621777 TI - Differential effects of regulatory T cells on the initiation and regression of atherosclerosis. AB - OBJECTIVE: Regulatory T cells (Tregs) play an important role in the regulation of T cell-mediated immune responses through suppression of T cell proliferation and cytokine production. In atherosclerosis, a chronic autoimmune-like disease, an imbalance between pro-inflammatory cells (Th1/Th2) and anti-inflammatory cells (Tregs) exists. Therefore, increased Treg numbers may be beneficial for patients suffering from atherosclerosis. In the present study, we determined the effect of a vast expansion of Tregs on the initiation and regression of well-established lesions. METHODS AND RESULTS: For in vivo Treg expansion, LDL receptor deficient (LDLr(-/-)) mice received repeated intraperitoneal injections of a complex of IL 2 and anti-IL-2 mAb. This resulted in a 10-fold increase in CD4(+)CD25(hi)Foxp3(+) T cells, which potently suppressed effector T cells ex vivo. During initial atherosclerosis, IL-2 complex treatment of LDLr(-/-) mice fed a Western-type diet reduced atherosclerotic lesion formation by 39%. The effect on pre-existing lesions was assessed by combining IL-2 complex treatment with a vigorous lowering of blood lipid levels in LDLr(-/-) mice. This did not induce regression of atherosclerosis, but significantly enhanced lesion stability. CONCLUSION: Our data show differential roles for Tregs during atherosclerosis: Tregs suppress inflammatory responses and attenuate initial atherosclerosis development, while during regression Tregs can improve stabilization of the atherosclerotic lesions. PMID- 21621778 TI - Arterial stiffness, pulse pressure, and cardiovascular disease-is it possible to break the vicious circle? AB - Hypertension is strongly associated with cardio/cerebrovascular diseases, e.g. myocardial infarction, stroke, and heart failure, which are main causes of cardiovascular morbidity and mortality. In hypertensive subjects, cardiovascular risk reduction is mainly associated with reduction in brachial systolic blood pressure (SBP). As it was shown in controlled and long-term therapeutic trials, it is possible to produce a selective SBP reduction through a specific "de stiffening" strategy. This means that SBP reduction is obtained independently of mean arterial pressure change, using a significant and selective reduction of wave reflections and/or aortic stiffness. The procedure is especially effective in decreasing central systolic and pulse pressures, which were shown to be major determinants of long-term outcome. As some concerns associated with decreasing in diastolic blood pressure to low values (so called "J-curve" phenomenon) have been raised recently the de-stiffening strategy appears to be especially attractive. Most of the protocols used to de-stiffen large arteries required the administration of a renin-angiotensin-aldosterone system inhibitor, which frequently was associated with a diuretic and/or a calcium antagonist, but not with a classic beta-blocker. These protocols were evaluated in randomized controlled trials and showed significant reduction in cardiovascular risk, particularly in comparison with beta-blockers. PMID- 21621779 TI - Rapid magnetic solid phase extraction with in situ derivatization of methylmercury in seawater by Fe3O4/polyaniline nanoparticle. AB - A new Fe(3)O(4)/polyaniline nanoparticle (PANI) material has been successfully developed as magnetic solid-phase extraction sorbent in dispersion mode for the determination of methylmercury (MeHg) in aqueous samples, via quantification by gas chromatography/mass spectrometry (GC-MS). The resultant core-shell magnetic solid-phase extraction nanoparticle (MSPE-NP) sorbent was characterized by scanning electron microscopy (SEM), energy dispersive X-ray spectrometry (EDS) and Fourier transform-infrared (FTIR) spectroscopy. Fe(3)O(4)/PANI composites showed fibrous structure with diameters between 50 and 100 nm for fibers. The MSPE-NP process involved the dispersion of the Fe(3)O(4)/PANI nanoparticles in water samples with sonication, followed by magnetic aided retrieval of the sorbent and then, solvent (hexane) desorption of extracted MeHg for GC-MS analysis. The extraction, derivatization and adsorption conditions were optimized by selecting the appropriate extraction parameters including the amount of sorbent, extraction time, derivatizing reagent volume and extraction solvent. The calibration graph was linear in the concentration range of 0.5-300 ng mL(-1) (R(2)>0.993) with detection limit of 0.1 ng mL(-1) (n=3), while the repeatability was 4.1% (n=5). Enrichment factor was obtained as 91. Seawater sample was analyzed as real sample and good recoveries (>98%) were obtained at different spiked values. PMID- 21621780 TI - Enhanced monitoring of biopharmaceutical product purity using liquid chromatography-mass spectrometry. AB - LC-MS is a widely used technique for impurity detection and identification. It is very informative and generates huge amounts of data. However, the relevant chemical information may not be directly accessible from the raw data map, particularly in reference to applications where unknown impurities are to be detected. This study demonstrates that multivariate statistical process control (MSPC) based on principal component analysis (PCA) in conjunction with multiple testing is very powerful for comprehensive monitoring and detection of an unknown and co-eluting impurity measured with liquid chromatography-mass spectrometry (LC MS). It is demonstrated how a spiked impurity present at low concentrations (0.05% (w/w)) is detected and further how the contribution plot provides clear diagnostics of the unknown impurity. This tool makes a fully automatic monitoring of LC-MS data possible, where only relevant areas in the LC-MS data are highlighted for further interpretation. PMID- 21621781 TI - Enthalpy of adsorption for hydrocarbons on concrete by inverse gas chromatography. AB - Enthalpies of adsorption, DeltaH(a), are reported for several light hydrocarbons on normal construction concrete. DeltaH(a), which are a measure of the adhesion strength of a molecule on a surface, were determined by gas-solid chromatography with a packed column containing 60-80 mesh concrete particles. With this approach, the specific retention volume for a compound is measured as a function of temperature, and these data are used to calculate DeltaH(a). For the hydrocarbons studied, we found that DeltaH(a) was relatively large for unsaturated hydrocarbons. These are the first determinations of DeltaH(a) of hydrocarbons on construction concrete, but useful comparisons with other ionic solids such as clays can be made. PMID- 21621782 TI - Hydrazone-based ligands for micro-solid phase extraction-high performance liquid chromatographic determination of biogenic amines in orange juice. AB - Eight hydrazone-based ligands were synthesized, trapped in a silica sol-gel matrix, and were subsequently used in the micro-solid phase extraction (MU-SPE) of biogenic amines (BAs). The BAs investigated were tryptamine, phenylethylamine, putrescine, histamine, tyramine and spermidine. Prior to the extraction, dansyl chloride was added to the samples which were heated to 70 degrees C for 10 min. The samples were extracted with MU-SPE, after which analytes were desorbed using acetonitrile via ultrasonication. The extracts were analysed by high performance liquid chromatography (HPLC) with ultraviolet detection. Of the eight ligands investigated as sorbents, benzophenone 2,4-dinitrophenylhydrazone was found to be the most promising. The enhanced pi-pi interaction between the analytes and the ligand facilitated the adsorption process. Under the most suitable extraction conditions, the method demonstrated good linearity with correlation coefficient of more than 0.985 over a concentration range of 1-50 MUg L(-1). Satisfactory repeatability with relative standard deviations of 7.43-11.30% (n=3) were obtained. Detection limits ranged from 3.8 to 31.3 ng L(-1). The MU-SPE method exhibited lower recoveries (71.5-87.4%) when compared to the solid phase extraction technique (79.7-95.0%), but enrichment factors of 94-460 were obtained. The proposed MU-SPE-HPLC method was applied to the determination of BAs in orange juice purchased from local supermarkets, with satisfactory results. The orange juices were characterized by the presence of relatively high levels of putrescine (range, 550-2210 MUg L(-1)) but tryptamine and phenylethylamine were not detected in any of the tested samples. PMID- 21621783 TI - Hydrophilic interaction liquid chromatography in food analysis. AB - The use of hydrophilic interaction liquid chromatography (HILIC) in food analysis in the last decade is reviewed. The HILIC mechanism is briefly discussed, but main emphasis is put on the use of HILIC for separation of different food matrices. The food matrices are divided into food of animal origin and related products, vegetables, fruits and related compounds, and other food-related matrices. A list on important applications is provided for each category including experimental conditions and a brief summary of the results. The 100 references included will provide the reader a comprehensive overview and insight into HILIC applications to food analysis. PMID- 21621784 TI - Selected tandem mass spectrometry ion monitoring for the fast identification of seafood species. AB - Selected tandem mass spectrometry (MS/MS) ion monitoring (SMIM) is the most suitable scanning mode to detect known peptides in complex samples when an ion trap mass spectrometer is the instrument used for the analysis. In this mode, the MS detector is programmed to perform continuous MS/MS scans on one or more selected precursors, either during a selected time interval, or along the whole chromatographic run. MS/MS spectra are recorded, so virtual multiple reaction monitoring chromatogram traces for the different fragment ions can be plotted. In this work, a shotgun proteomics approach was applied to the detection of previously characterized species-specific peptides from different seafood species. The proposed methodology makes use of high intensity focused ultrasound assisted trypsin digestion for ultra fast sample preparation, peptide separation and identification by reverse phase capillary LC coupled to an ion-trap working in the SMIM scanning mode. This methodology was applied to the differential classification of seven commercial, closely related, species of Decapoda shrimps proving to be an excellent tool for seafood product authentication, which may be used by fisheries and manufacturers to provide a fast and effective identification of the specimens, guaranteeing the quality and safety of foodstuffs to consumers. PMID- 21621785 TI - Using multiple short-end injections to develop fast electrophoretic separations- applications in iodide analysis. AB - The aim of this study was to develop a fast CE separation method by using multiple short-end injections in a capillary coated with quaternary ammonium chitosan (HACC), in order to determine the iodide content of pharmaceutical formulations. The BGE was composed of 20 mM tris(hydroxymethyl)aminomethane and 11 mM hydrochloric acid, at pH 8. The internal standard used was thiocyanate. Separations were performed in a fused silica capillary (32 cm total length, 8.5 cm effective length and 50 MUm i.d.) coated with HACC and direct UV detection at 220 nm. EOF was modified by flushing the capillary with polymeric solution, resulting in a semi-permanent coating of controlled and stable EOF. The EOF was anodic at pH 8. Different strategies, using single and multiple injection short end configurations, were studied to develop a CE method that resulted in a maximum number of iodide samples analyzed per hour: one plug and flush (Sflush) 35 samples/h, one plug without flush (SWflush) 76 samples/h, four plugs and flush (Mflush) 61 samples/h, and four plugs without flush (MWflush) 80 samples/h. Using the multiple injection configuration, it was possible to inject up to four plugs using spacer electrolytes with good separation efficiency and selectivity. The voltage application time needed to separate the eight peaks (iodide and thiocyanate) with MWflush was only 12s. The method was validated and samples were analyzed using MWflush. Good linearity (R(2)>0.999); a limit of detection 0.4 mg L(-1); intermediate precision better than 3.8% (peak area) and recovery in the range of 99-102% were obtained. PMID- 21621786 TI - Multi-mycotoxin analysis in eggs using a QuEChERS-based extraction procedure and ultra-high-pressure liquid chromatography coupled to triple quadrupole mass spectrometry. AB - A reliable and rapid method has been developed for the determination of 10 mycotoxins (beauvericin, enniatin A, A1, B1, citrinin, aflatoxin B1, B2, G1, G2 and ochratoxin A) in eggs at trace levels. Ultra-high-pressure liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) has been used for the analysis of these compounds in less than 7 min. Mycotoxins have been extracted from egg samples using a QuEChERS-based extraction procedure (Quick, Easy, Cheap, Effective, Rugged and Safe) without applying any further clean-up step. Extraction, chromatographic and detection conditions were optimised in order to increase sample throughput and sensitivity. Matrix-matched calibration was used for quantification. Blank samples were fortified at 10, 25, 50 and 100 MUg kg(-1), and recoveries ranged from 70% to 110%, except for ochratoxin A and aflatoxin G1 at 10 MUg kg(-1), and aflatoxin G2 at 50 MUg kg(-1). Relative standard deviations were lower than 25% in all the cases. Limits of detection ranged from 0.5 MUg kg(-1) (for aflatoxins B1, B2 and G1) to 5 MUg kg(-1) (for enniatin A, citrinin and ochratoxin A) and limits of quantification ranged from 1 MUg kg(-1) (for aflatoxins B1, B2 and G1) to 10 MUg kg(-1) (for enniatin A, citrinin and ochratoxin A). Seven samples were analyzed and aflatoxins B1, B2, G1, G2, and beauvericin were detected at trace levels. PMID- 21621787 TI - Accurate measurements of peak variances: importance of this accuracy in the determination of the true corrected plate heights of chromatographic columns. AB - The true efficiency of a column is derived from the differences between the variances of the peak profiles of the same compound recorded in the presence and the absence of the chromatographic column. These variances are usually derived using one of three methods: (1) the retention time of the peak apex and its half height width; (2) the moments of the best fit between the experimental data and a hybrid response function, e.g., an exponentially convoluted Gaussian; or (3) the exact moments of the experimental band profiles. Comparisons of the results of these methods show that the first method is always inaccurate because all the band profiles recorded are strongly tailing. The peak fit method is accurate only for 4.6mm I.D. columns operated with instruments having low extra-column volume but fails for short narrow-bore columns due to the severe tailing of peaks passing through the complex channels of the extra-column volumes and to the inaccuracies in the fit of experimental data to the selected function. Although far better, the moment method may be inaccurate when the zero dead volume union used to measure the extra-column peak variances has a higher permeability than the column, causing the upstream part of the instrument to operate under comparatively low pressures. PMID- 21621788 TI - Derivatization-independent cholesterol analysis in crude lipid extracts by liquid chromatography/mass spectrometry: applications to a rabbit model for atherosclerosis. AB - Direct measurement of various sterols in crude lipid extracts in a single experiment from limited biological samples is challenging. Current mass spectrometry (MS) based approaches usually require chemical derivatization before subjecting to MS analysis. Here, we present a derivatization-independent method for analyzing various sterols, including cholesterol and its congeners, using liquid chromatography and atmospheric pressure chemical ionization mass spectrometry. Based on the specific tandem mass spectrometry pattern of cholesterol, multiple reaction monitoring (MRM) transitions were used to quantify free cholesterol and its fatty acyl esters. Several cholesterol oxidation products could also be measured using the upfront liquid chromatography separation and specific MRM transitions. The method was validated alongside established enzymatic assays in measuring total cholesterol. As a proof of concept, we analyzed plasma sterols in rabbits administrated with a high cholesterol diet (HCD) which is a classical atherosclerotic model. Free cholesterol, cholesterol esters, 7-hydroxycholesterol, and 7-ketocholesterol were elevated in plasma of rabbits on HCD. This method could also serve as an excellent tool for quantitative analysis of other sterols such as ergosterol and sitosterol in other organisms beside mammalian. In Saccharomyces cerevisiae, our results indicated dramatic increases of the ratio of ergosterol esters to free ergosterol in both yeh2Delta and tgl1Delta cells, which are consistent with the function of the respective enzymes. PMID- 21621789 TI - The effect of copper on the precipitation of scorodite (FeAsO4.2H2O) under hydrothermal conditions: evidence for a hydrated copper containing ferric arsenate sulfate-short lived intermediate. AB - The effect of copper sulfate on scorodite precipitation and its mechanism of formation at 150 degrees C was investigated. Scorodite was determined to be the dominant phase formed under all conditions explored (0.61 < Fe(III)/As(V) < 1.87, 0.27-0.30 M Fe(SO(4))(1.5), 0-0.3 M CuSO(4), 0-0.3 M MgSO(4), at 2.5 h and 150 degrees C). The produced scorodite was found to incorporate up to 5% SO(4) and <=1% Cu or Mg in its structure. The precipitation of scorodite was stoichiometric, i.e. the Fe/As molar ratio in the solids was equal to one independent of the starting Fe/As ratio in the solution. The presence of excess ferric sulfate in the initial solution (Fe/As>1) was found to slow down the ordering of the H-bond structure in scorodite. Precipitation under equimolar concentrations (As = Fe = Cu = 0.3 M), short times and lower temperatures (30-70 min and 90-130 degrees C) revealed the formation of a Cu-Fe-AsO(4)-SO(4)-H(2)O short lived gelatinous intermediate that closely resembled the basic ferric arsenate sulfate (BFAS) type of phase, before ultimately converting fully to the most stable scorodite phase (96 min and 138 degrees C). This phase transition has been traced throughout the reaction via elemental (ICP-AES, XPS), structural (PXRD, TEM) and molecular (ATR-IR, Raman) analysis. ATR-IR investigation of an arsenic containing industrial residue produced during pressure leaching of a copper concentrate (1 h and 150 degrees C) found evidence of the formation of an arsenate mineral form resembling the intermediate basic ferric arsenate sulfate phase. PMID- 21621790 TI - Enzymatic tailoring for precise control of plasmonic resonance absorbance of gold nanoparticle assemblies. AB - We report an enzymatic method to control the plasmon resonance absorbance of gold nanoparticle (AuNP) arrays assembled on hyaluronic acids. While multiple electrostatic interactions between cysteamine on the AuNPs and the carboxylic acid residues in the whole intact hyaluronic acid induced the formation of large aggregates, precise control of the plasmon absorbance was possible by tailoring the size of the bio-polymeric templates with hyaluronidase, almost over the entire range of the resonant coupling wavelengths. It was possible to precisely tune the position of the second plasmon absorbance by manipulating the amount of the template and the enzymatic hydrolysis time. Finally, we were able to produce a chain-like array of AuNPs, which was nearly one dimensional, with a maximum shift of up to 189 nm in the plasmon absorbance at the optimal hydrolysis time of the templates. This enzymatic method can be used as a useful tool to tailor the plasmonic properties of the nanostructures required for specific applications. PMID- 21621791 TI - Effect of Al-doping on the structure and optical properties of electrospun zinc oxide nanofiber films. AB - Electrospun ZnO precursor nanofibers of average diameters 122+/-64 nm, 117+/-44 nm and 110+/-39 nm were fabricated by controlling the Al concentration of a polymeric solution. The resulting nanofibers were characterized by the XRD, SEM, EDS, TEM, XPS and PL. The electrospun Al-doped ZnO nanofiber films were polycrystalline and composed of densely packed grains, with crystallite size ranging from 28.7 nm, 25.7 nm, 25.4 nm to 20.4 nm corresponding to the atomic concentration of aluminum from 0, 1.6, 2.5 to 5.8 at.%. The incorporation of aluminum resulted in a decrease trend in the grain size and lattice parameter of the ZnO nanofiber films. The room temperature PL spectra of all samples show three different emissions, including UV (ultraviolet) emission with an obvious blue shift, Vis (visible) emission and NIR (near infrared) emission, the intensity of which decreases monotonically as the doping concentration is increased except for the highest doping level. The impurity content correlates with changes in the PL spectra, and the appropriate Al doping can improve the optical properties of ZnO nanofibers. The small size effect and Al-doping or the impurity incorporation should be responsible for the blue shift observation in Al doped ZnO nanofiber films. PMID- 21621792 TI - Bile acid alkylamide derivatives as low molecular weight organogelators: systematic gelation studies and qualitative structural analysis of the systems. AB - A series of amino- and hydroxyalkyl amides of bile acids have been synthesized and characterized by Fourier transform infrared spectroscopy (FTIR), (1)H and (13)C nuclear magnetic resonance spectroscopy (NMR), as well as electrospray ionization mass spectrometry (ESI-MS) measurements. The ability of the synthesized molecules to promote gel formation was systematically investigated. Out of 396 combinations formed by 11 compounds and 36 different solvents, 22 gel containing systems were obtained with 1% (w/v) gelator concentration. Apart from one exception, the gelator compounds were lithocholic acid derivatives. This challenges the general trend of bile acid-based physical gelators, according to which the gelation ability of lithocholic acid derivatives is poor. A correlation between the values of Kamlet-Taft parameters and solvent preferences for gelators was observed. The morphologies of the solid and gel structures studied with scanning electron microscopy (SEM) showed variability from fibers to spherical microscale aggregates, the latter of which are unique among bile acid-based organogels. The gels exhibited more complex behavior than was previously established with bile acid derivatives, judging by the microscale diversity present in gelating and non-gelating systems and the tendency for polymorphism. This study underlines the importance of both the molecular and colloidal scale aspects of the gelation phenomenon. PMID- 21621793 TI - Influence of a nanorod molecular layer on the biological activity of neuronal cells. A semiclassical model for complex solid/liquid interfaces with carbon nanotubes. AB - A general account of electric effects is given for a biological phase interacting with a nanorod molecular layer by means of the formed hard-soft and solid-liquid interfaces. In particular, the frequency enhancement previously detected for the spontaneous activity of neuronal cultures interfaced with carbon nanotubes is quantitatively explained upon a quantum/semiclassical description, where the duration of a biological signal is viewed as the (average) lifetime of a decaying state (or population of states), and the effect of the carbon phase as a linewidth broadening. Four contributions were principally accounted for, one biological, for the synaptic strength, one electrochemical, for the overall capacitance increase implied by the nanotube double layers, one geometric, for the typical scales ruling the electron and ion conduction mechanisms, and one electromagnetic-like, translating the membrane polarization changes. These calculations predict an enhancement factor equal on average to ?6.39, against a former experimental value ?6.08. PMID- 21621794 TI - Comparative studies on brij reverse micelles prepared in benzene/surfactant/ethylammonium nitrate systems: effect of head group size and polarity of the hydrocarbon chain. AB - Nonaqueous reverse micelles of brij surfactants are prepared in benzene and ethylammonium nitrate (EAN). The effect of polar head group bulk on reverse micellar size was studied with brij-52, brij-56 and brij-58 whereas the effect of polarity of hydrocarbon chain was investigated taking brij-52 and brij-93 with varying W(s) (W(s)=[EAN]/[surfactant]). Dynamic light scattering (DLS) has been employed to reveal the size and shape of the reverse micelles. Micropolarities of these reverse micelles were investigated by visible spectroscopy using methylene blue (MB) and methyl orange (MO) as molecular optical probes. It has been revealed from the experimental results that with increase in polar head group size reverse micellar size increases. Moreover, it is also observed that with increasing polarity of the hydrocarbon chain the average size of the reverse micelles decreases. It can be concluded that polar head group size and polarity of hydrocarbon chain play important roles in determining reverse micellar size of the brij surfactants apart from the W(s) ratio, nature of the solvent medium, and concentration of the surfactants. PMID- 21621795 TI - Drug-induced dysimmune demyelinating neuropathies. AB - Drug-induced peripheral neurotoxicity usually manifests as a length-dependent, "dying back" axonal, predominantly sensory polyneuropathy. Rarely, immune mediated demyelinating neuropathies occur during initial or maintenance treatment with immunomodulatory, immunosuppressive or antineoplastic agents. Medication induced immune perturbation presumably triggers a dysimmune attack directed at unidentified peripheral nerve myelin epitopes; true peripheral nerve toxicity (i.e., dependent on accumulative dose or serum level) plays no identified role. The mechanisms that underlie a paradoxical and unpredictable immune exacerbation are unclear, and may depend on patient age, drug dosage and schedule, time of treatment relative to disease course, and host genetic factors. Suspicion and recognition of a non-toxic, immune-mediated demyelinating process has management (targeted immunotherapy) and prognostic (mostly favorable) implications. PMID- 21621796 TI - Anxiety and depression in multiple sclerosis patients around diagnosis. AB - OBJECTIVES: To prospectively identify anxiety and depressive symptoms, and their predictors, during the multiple sclerosis (MS) peridiagnostic period. METHODS: The Hospital Anxiety and Depression Scale (HADS) was administered during diagnostic workup (baseline), and one and six months after diagnosis disclosure, to SIMS-Trial participants (ISRCTN81072971). RESULTS: Of 197 screened patients, 120 (61%) were diagnosed with MS. At baseline, median HADS anxiety (HADS-A) score was 7.0 (interquartile range [IQR] 5.0-9.5), >=8 (anxiety cut-off) in 43% (95% confidence interval [CI] 34%-52%). Median HADS depression (HADS-D) was 3.0 (IQR 1.0-5.0), >=8 (depression cut-off) in 11% (95% CI 5%-16%). Independent predictors of anxiety were female sex (odds ratio [OR] 2.8, 95% CI 1.1-7.2) and HADS-D score (OR 20.8, 95% CI 2.5-175.5). The only predictor of depressive symptoms was HADS-A score (OR 20.0, 95% CI 2.8-260.9). Anxiety symptoms had decreased slightly but significantly (p<0.001) at six months. Depressive symptoms remained low. CONCLUSIONS: Anxiety was prominent in the period surrounding MS diagnosis disclosure, particularly in women. A slight but significant reduction occurred six months after diagnosis disclosure. Depressive symptoms were less common and stable over time. In addition to sex, depressive symptoms were the only variable independently associated with anxiety. PMID- 21621797 TI - Lesion patterns and mechanism of cerebral infarction caused by severe atherosclerotic intracranial internal carotid artery stenosis. AB - BACKGROUND AND PURPOSE: The lesion patterns and mechanisms of ischemic stroke caused by extracranial internal carotid artery (EICA) stenosis are well understood. The prognosis of intracranial internal carotid artery (IICA) stenosis is very poor. However, little is known about lesion patterns and mechanisms of cerebral infarcts caused by IICA stenosis. The objective of this study was to investigate the lesion patterns and mechanisms of infarcts produced by severe IICA stenosis and compare it with that produced by severe EICA stenosis. METHODS: We recruited 62 patients with acute cerebral infarcts who fulfilled the following criteria: 1) lesions of acute infarcts were verified by diffusion-weighted imaging (DWI) performed within 1 week of stroke onset; 2) infarct lesions were located within the territory of unilateral internal carotid artery (ICA); 3) infarcts were definitely caused by severe atherosclerosis stenosis (stenosis rate of 70%-99%) of the ipsilateral IICA or EICA, with all other potential causes of stroke being entirely excluded. According to the distributions of infarct lesions in the arterial territory of the ICA displayed on DWI, lesion patterns were classified as either 1) single infarct [perforating artery infarct (PAI), pial artery infarct (PI) or border-zone infarct (BZI)], or 2) multiple infarcts (a combination of types described above). RESULTS: There were 29 patients with ischemic stroke caused by severe IICA stenosis, and 33 patients with stroke caused by severe EICA stenosis. Single BZI (14/29, P=0.015), and infarcts involving the border zone (19/29, P=0.021) or the internal border zone (13/29, P=0.013) were identified more often in patients with IICA stenosis compared to those with EICA stenosis. PI and/or PAI (22/32, P=0.021) were identified more often in patients with EICA stenosis. CONCLUSIONS: Lesion patterns in patients with severe IICA stenosis were different from those with severe EICA stenosis. The hypoperfusion mechanism leading to BZI was more important for patients with severe IICA stenosis than for those with severe EICA stenosis. Embolisms leading to PI and/or PAI were more important for patients with severe EICA stenosis. PMID- 21621798 TI - Long-term trends in benthic habitat quality as determined by Multivariate AMBI and Infaunal Quality Index in relation to natural variability: a case study in Kinsale Harbour, south coast of Ireland. AB - Benthic Ecological Quality Ratios (EQR) are important tools for assessing the ecological status of coastal and transitional water bodies. Here, we use spatial and time-series data from Kinsale Harbour, Ireland to examine the effects of sample processing methodologies on the outputs of two EQRs: Multivariate AMBI (M AMBI) and Infaunal Quality Index (IQI). Both EQRs were robust to changes in sieve size from 1mm to 0.5mm, and to changes in the taxa identified in spatial calibration. Both EQRs classified habitat quality in Kinsale as generally Good or High with no evidence of significant change over the time series (1981-2006). IQI classified the ecological status as higher than M-AMBI. There was a significant relationship between IQI and M-AMBI in spatial calibration, but no significant relationship between them in time series. Further research into the behaviour of EQRs in relation to natural variability over long time-scales is needed to discriminate anthropogenic impacts reliably. PMID- 21621799 TI - [Adverse effects of antiarrhythmic drugs in patients with atrial fibrillation in primary care]. AB - BACKGROUND AND OBJECTIVE: To determine the adverse effects (AE) of antiarrhythmic drugs used to treat atrial fibrillation (AF) in routine clinical practice. PATIENTS AND METHODS: A retrospective, observational, population-based study of all patients diagnosed of AF requiring long-term treatment with antiarrhythmic drugs. We performed also a descriptive analysis of the type of EA, and their management (diagnostic techniques, laboratory tests, drug treatment, number of visits to health services, emergency visits, hospitalizations and surgeries). RESULTS: We included 400 patients. Overall, 688 treatments were started, 34 of which (4.9%) were implied in 32 EA in 30 patients. The most frequent EA were hypothyroidism (28.1%), hyperthyroidism (25%) and bradycardia (9.38%). In absolute numbers, class III drugs (Vaughan-Williams classification) were associated with a higher number of AE (p=0.001). The number of AE/100 patients year was: class I 0.42 (0.01-1.15%), class II 2.55 (0.68-5.13%), class III 4.70 (2.98-6.64%), class IV 1.18 (0.14-2.80%), digoxin 0.41 (0.08-1.19). Individually, the drugs with a higher rate were sotalol, bisoprolol and amiodarone (5.70, 5.37 and 4.59 respectively). 93.75% of EA required visits to health services, 59.38% laboratory tests and 56.25% diagnostic techniques. The most frequent visits were scheduled for primary care (75%), followed by scheduled visits to outpatient cardiology (21.9%). 71.9% of EA were resolved without any sequel and one patient died. CONCLUSIONS: Antiarrhythmic drugs for the management of AF are associated with the presence of AE, mainly class III drugs. In most cases they are not serious, but imply important health resources. PMID- 21621800 TI - [Presacral extramedullary hematopoiesis secondary to congenital heart disease]. PMID- 21621801 TI - The effect of two energy-restricted diets, a low-fructose diet versus a moderate natural fructose diet, on weight loss and metabolic syndrome parameters: a randomized controlled trial. AB - One of the proposed causes of obesity and metabolic syndrome is the excessive intake of products containing added sugars, in particular, fructose. Although the ability of excessive intake of fructose to induce metabolic syndrome is mounting, to date, no study has addressed whether a diet specifically lowering fructose but not total carbohydrates can reduce features of metabolic syndrome. A total of 131 patients were randomized to compare the short-term effects of 2 energy-restricted diets-a low-fructose diet vs a moderate natural fructose diet-on weight loss and metabolic syndrome parameters. Patients were randomized to receive 1500, 1800, or 2000 cal diets according to sex, age, and height. Because natural fructose might be differently absorbed compared with fructose from added sugars, we randomized obese subjects to either a low-fructose diet (<20 g/d) or a moderate-fructose diet with natural fruit supplements (50-70 g/d) and compared the effects of both diets on the primary outcome of weight loss in a 6-week follow-up period. Blood pressure, lipid profile, serum glucose, insulin resistance, uric acid, soluble intercellular adhesion molecule-1, and quality of life scores were included as secondary outcomes. One hundred two (78%) of the 131 participants were women, mean age was 38.8 +/- 8.8 years, and the mean body mass index was 32.4 +/- 4.5 kg/m(2). Each intervention diet was associated with significant weight loss compared with baseline. Weight loss was higher in the moderate natural fructose group (4.19 +/- 0.30 kg) than the low-fructose group (2.83 +/- 0.29 kg) (P = .0016). Compared with baseline, each intervention diet was associated with significant improvement in secondary outcomes. Reduction of energy and added fructose intake may represent an important therapeutic target to reduce the frequency of obesity and diabetes. For weight loss achievement, an energy restricted moderate natural fructose diet was superior to a low-fructose diet. PMID- 21621802 TI - "Dinoflagellate Sterols" in marine diatoms. AB - Sterol compositions for three diatom species, recently shown to contain sterols with side chains typically found in dinoflagellates, were determined by HPLC and 1H NMR spectroscopic analyses. The centric diatom Triceratium dubium (=Biddulphia sp., CCMP 147) contained the highest percentage of 23-methylated sterols (37.2% (24R)-23-methylergosta-5,22-dienol), whereas the pennate diatom Delphineis sp. (CCMP 1095) contained the cyclopropyl sterol gorgosterol, as well as the 27 norsterol occelasterol. The sterol composition of Ditylum brightwellii (CCMP 358) was the most complex, containing Delta0- and Delta7-sterols, in addition to the predominant Delta5-sterols. A pair of previously unknown sterols, stigmasta 5,24,28-trienol and stigmasta-24,28-dienol, were detected in D. brightwellii and their structures were determined by NMR spectroscopic analysis and by synthesis of the former sterol from saringosterol. Also detected in D. brightwellii was the previously unknown 23-methylcholesta-7,22-dienol. PMID- 21621803 TI - Differential phenolic profiles in six African savanna woody species in relation to antiherbivore defense. AB - Low molecular weight phenolics are suggested to have a role in mediating diet selection in mammalian herbivores. However, very little is known about low molecular weight phenolic profiles of African savanna woody species. We determined low molecular weight phenolic profiles of six woody species with different life history, morphological and functional traits. We investigated interspecific phytochemical variation between species and found that: (1) related Acacia species were chemically dissimilar; (2) similarity percentage analysis revealed that Acacia grandicornuta was most dissimilar from other species and that the evergreen and unpalatable Euclea divinorum had a qualitatively similar chemical profile to the deciduous and palatable Acacia exuvialis and Combretum apiculatum; (3) C. apiculatum had the highest chemical diversity; (4) relative to spineless plants, spinescent plants contained significantly less HPLC phenolics and condensed tannins; and (5) the major quantitative difference between the evergreen and unpalatable E. divinorum and other species was its high myricitrin concentration. PMID- 21621804 TI - [How to read Radiologia today?]. PMID- 21621805 TI - [How to write an original article for a biomedical journal]. AB - Writing a scientific article is difficult, but skillful writing does not depend only on talent. This skill can be learned, and writing requires discipline and dedication. This article discusses the basic characteristics of the IMRaD (Introduction, Material and methods, Results, and Discussion) format and the contents of each section, as well as the characteristics of the title, abstract, and bibliographical references. It also provides a list of five characteristics that are essential for an original article published in a scientific journal: objective, permission of the ethics committee, a structured material and methods section, results that follow appropriately from the material and methods section, and a structured discussion. Furthermore, we provide a basic outline for the discussion section consisting of stating the most important results, developing a critical discussion of the results, stating and discussing the strengths and limitations of the study, and elaborating the final synthesis. PMID- 21621806 TI - Occurrence and functionality of cycle inhibiting factor, cytotoxic necrotising factors and cytolethal distending toxins in Escherichia coli isolated from calves and dogs in Italy. AB - Escherichia coli isolated from animals up to three months of age, with diarrhea (255 calves and 29 dogs (pups)), without diarrhea (21 calves and 11 pups, used as controls), and 58 adult dogs with cystitis were tested to investigate the occurrence and functional expression of cyclomodulins cycle inhibiting factor (CIF), cytotoxic necrotizing factors (CNFs) and cytolethal distending toxins (CDTs). In cyclomodulin-positive isolates the association was assessed with other virulence genotypes and phylogenetic groups. Of 374 E. coli isolates, 80 (21.4%) were positive for at least one cyclomodulin and 14 of the latter (3.7%) showed different combinations of more than one. cif-positive isolates showed a low number of additional virulence factors, and were commonly associated with phylogroup B1, while cnf- and cdt-positive isolates, harboring many extraintestinal virulence factors, belonged to phylogroups B2 and D. Almost all isolates showed an irreversible cytopathic effect (CPE), displaying functionality of cyclomodulins. Five isolates that presented a mutation of cif were CPE negative. PMID- 21621807 TI - Magnitude and causes of childhood blindness and severe visual impairment in Sekoru District, Southwest Ethiopia: a survey using the key informant method. AB - There are very few population-based data on childhood blindness in the Horn of Africa. In Sekoru district, Ethiopia, we performed a whole-of-population cross sectional survey using the key informant method to determine the magnitude and causes of childhood blindness (presenting visual acuity <3/60 in the better eye) and severe visual impairment (SVI; presenting visual acuity <6/60 but >=3/60 in the better eye). Forty-two key informants were trained to identify blind/SVI children aged <16 years. Identified children were examined by an ophthalmologist. Causes of blindness/SVI were established by clinical examination. Key informants visited 94% of 22,666 households, identifying 112 children, of whom 36 (32%) were confirmed to be blind/SVI by the ophthalmologist. Twenty (56%) of the 36 were male. The mean age was 10.7 years. The district prevalence of childhood blindness/SVI was 0.062% (95% CI 0.042-0.082%). Lens-related abnormalities (mainly congenital cataract) were the commonest causes. Avoidable causes of blindness accounted for 89% of cases. Paediatric ophthalmic services are required here, as elsewhere in rural Ethiopia. Assessment by key informants has an acceptable positive predictive value in this setting. This method warrants further evaluation as a tool for measuring the prevalence of other rare conditions in rural and remote populations. PMID- 21621808 TI - A twisted mess. AB - A 12-year-old girl presented with painless decreased vision in the left eye. Dilated fundus examination was consistent with Wyburn-Mason syndrome, and subsequent neuroimaging demonstrated a retro-orbital arteriovenous malformation. The etiology of the vision loss is discussed, as well as management options of Wyburn-Mason syndrome. PMID- 21621809 TI - Bilateral optic disk swelling plus. AB - A 64-year-old woman presented with bilateral optic neuropathy leading to a diagnosis of Sjogren syndrome. She improved with high-dose corticosteroids and oral azathioprine and was subsequently found to have asymptomatic bilateral iridocyclitis. Although central nervous system manifestations of Sjogren syndrome are documented in the literature, they are not widely recognized in clinical practice. Associated optic neuritis often mimics demyelinating disease such as multiple sclerosis. Treatment of CNS disease related to Sjogren syndrome is highly controversial. Uveitis is an uncommon finding associated with Sjogren syndrome. PMID- 21621810 TI - Identification of estrogen receptor agonists in sediments from Wenyu River, Beijing, China. AB - Assignment of ecological impacts of contamination to specific classes of contaminants is a prerequisite for risk assessment and remediation. In this study, the combination of polarity-based fractionation, two-hybrid yeast bioassay, and chemical analysis were used to evaluate and identify estrogen receptor agonists (ER-agonists) in sediments from Wenyu River, Beijing, China. By bioassay, organic raw sediment extracts could induce significant estrogenicity and the bioassay-derived 17beta-estradiol equivalents (EEQs) of raw extracts (EEQ(raw)s) ranged from 0.8 to 19.8 ng/g dry weight. By polarity-based fractionation, the raw extracts were separated into three fractions, i.e. non polar, moderately polar, and polar fractions, which were subjected to bioassay and chemical analysis. The highest estrogenicity was observed in the polar fraction, which accounted for more than 78% of the total. The medium polar fraction contains PAHs and OCPs, and the estrogenic activities in this fraction contributed 3%-12% of the total in raw extract. An estrogenic activity of non polarity fraction was negligible in compare to other two fractions. By chemical analysis and toxic equivalent calculation, major part of the estrogenicity in polar fraction could be attributed to six natural/synthetic estrogens (16%-63%), i.e. 17beta-estradiol, estrone, estriol, 17alpha-ethynylestradiol, diethylstilbestrol, and beta-estradiol-17-valerate, and to nonylphenols (26% 55%). The proposed approach has been successfully used for characterization of ER agonists in this case study. PMID- 21621811 TI - Synthesis and biological activity of derivatives of tetrahydroacridine as acetylcholinesterase inhibitors. AB - Current state of medical sciences does not allow to treatment neurodegenerative diseases such as Alzheimer's disease (AD). At present treatment of AD is severely restricted. The main class of medicines which are applied in AD is acetylcholinesterase inhibitors (AChEIs) like tacrine, donepezil, galantamine and rivastigmine that do not contribute to significant and long-term improvement in cognitive and behavioural functions. In this work, we report synthesis and biological evaluation of new hybrids of tacrine-6-hydrazinonicotinamide. The synthesis was based on the condensation reaction between tacrine derivatives and the hydrazine nicotinate moiety (HYNIC). All obtained compounds present affinity for both cholinesterases and are characterized by high selectivity in relation to butyrylcholinesterase (BChE). PMID- 21621812 TI - Aqueous photoreduction of oxidized mercury species in presence of selected alkanethiols. AB - Mercury is a global environmental contaminant with severe toxicity impact. The chemical processes resulting in the transformation of oxidized mercury species (Hg2+) to elemental mercury (Hg0), greatly affects the fate and transport of mercury in the natural environment. We hereby provide the first study on the photochemistry of Hg2+ with selected alkanethiols (R-SH) as model compounds to represent thiols and thiol-type binding sites on humic substances in natural waters because of the common sulfhydryl functional group (-SH). Kinetic studies were performed using cold vapor atomic fluorescence spectroscopy (CVAFS), the formation of Hg2+-thiol complexes (Hg(SR)2) were confirmed by UV-visible spectrometry and Atmospheric Pressure Chemical Ionization-Mass Spectrometry (APCI MS), and the reaction products were analyzed using Electron Impact-Mass Spectrometry (EI-MS) and Solid Phase Microextraction coupled with Gas Chromatography-Mass Spectrometry (SPME/GC-MS). Our results indicated that the photoreduction of Hg2+ by selected alkanethiols may be mediated by Hg2+-thiol complexes to produce Hg0. Under our experimental conditions, the apparent first order rate constants obtained for 1-propanethiol, 1-butanethiol, and 1 pentanethiol were (2.0+/-0.2)*10(-7) s(-1), (1.4+/-0.1)*10(-7) s(-1), (8.3+/ 0.5)*10(-8) s(-1), respectively. The effects of ionic strength, dissolved oxygen or chloride ion on reaction rates were found to be minimal under our experimental conditions. The identified products of the reaction between oxidized mercury species with selected alkanethiols (C3-C5) were Hg0 and disulfides (RS-SR). The potential environmental implications are herein discussed. PMID- 21621813 TI - Biodegradation of aromatic hydrocarbons by Haloarchaea and their use for the reduction of the chemical oxygen demand of hypersaline petroleum produced water. AB - Ten halophilic Archaea (Haloarchaea) strains able to degrade aromatic compounds were isolated from five hypersaline locations; salt marshes in the Uyuni salt flats in Bolivia, crystallizer ponds in Chile and Cabo Rojo (Puerto Rico), and sabkhas (salt flats) in the Persian Gulf (Saudi Arabia) and the Dead Sea (Israel and Jordan). Phylogenetic identification of the isolates was determined by 16S rRNA gene sequence analysis. The isolated Haloarchaea strains were able to grow on a mixture of benzoic acid, p-hydroxybenzoic acid, and salicylic acid (1.5mM each) and a mixture of the polycyclic aromatic hydrocarbons, naphthalene, anthracene, phenanthrene, pyrene and benzo[a]anthracene (0.3mM each). Evaluation of the extent of degradation of the mixed aromatic hydrocarbons demonstrated that the isolates could degrade these compounds in hypersaline media containing 20% NaCl. The strains were shown to reduce the COD of hypersaline crude oil reservoir produced waters significantly beyond that achieved using standard hydrogen peroxide treatment alone. PMID- 21621814 TI - Long-term sodium and chloride surface water exports from the Dallas/Fort Worth region. AB - Sodium and chloride in surface water are typically related to urbanization and population density and can have a significant impact on drinking water sources and the subsequent salinity of aquatic ecosystems. While the majority of research has focused on the impact of deicing salts on urban surface waters in colder climates, the effect of urbanization on sodium and chloride concentrations has been found to occur in warmer climates. This study investigated long-term exports of sodium and chloride from watersheds with increasing urbanization in the humid subtropical Dallas-Fort Worth region. We compared exports to characteristics of urbanization: urban land cover, impervious surface area, and calculated contributions from wastewater discharges. Long-term data (1980-2008) were obtained from five USGS gages located in and around the cities. Exports were calculated by regression analysis between concentrations and discharge and normalized for time and the watershed area. Grab samples were collected from June 2009 to May 2010 and sodium and chloride concentrations quantified. Our results show a strong positive relationship between the mean annual sodium and chloride exports from each watershed and the percent urban land cover and impervious surface area. Long-term increases in sodium and chloride fluxes were found for the three watersheds with the highest percentage of urban land cover. The single largest contributor was wastewater effluent that was estimated to contribute approximately half of the total loads in the three urbanized watersheds. Atmospheric deposition and deicing salts accounted for small amounts of the total export for urbanized watersheds. The source of the remaining salt load is still unknown and may be a combination of non-point sources. Estimates of urban salt exports were similar to estimates from northern watersheds affected by deicing salts. PMID- 21621815 TI - Effects of gamma-sterilization on DOC, uranium and arsenic remobilization from organic and microbial rich stream sediments. AB - Organic-rich sediments are known to be effective accumulators for uranium and arsenic. Much is known about the capacity for metal or metalloid fixation by microbes and organic compounds as well as inorganic sediment particles. Experiments investigating the effect of microbes on the process of metal fixation in sediments require sterilized sediments as control treatment which is often realized by gamma-sterilization. Only few studies show that gamma-sterilization has an effect on the remobilization of metal and metalloids and on their physico chemical properties. These studies deal with sediments with negligible organic content whereas almost nothing is known about organic-rich sediments including a probably high microbial activity. In view of this, we investigated the effect of gamma-sterilization of organic-rich sediments on uranium and arsenic fixation and release. After ten days within an exposure experiment we found a significant higher remobilization of uranium and arsenic in sterile compared to unsterile treatments. In line with these findings the content of dissolved organic carbon (DOC), manganese, and iron increased to even significantly higher concentration in the sterile compared to unsterile treatment. Gamma-sterilization seems to change the physico-chemical properties of organic-rich sediments. Microbial activity is effectively eliminated. From increased DOC concentrations in overlaying water it is concluded that microbes are eventually killed with leaching of cellular compounds in the overlaying water. This decreases the adsorption capacity of the sediment and leads to enhanced uranium and arsenic remobilization. PMID- 21621816 TI - Atmospheric emissions estimation of Hg, As, and Se from coal-fired power plants in China, 2007. AB - Over half of coal in China is burned directly by power plants, becoming an important source of hazardous trace element emissions, such as mercury (Hg), arsenic (As), and selenium (Se), etc. Based on coal consumption by each power plant, emission factors classified by different boiler patterns and air pollution control devices configuration, atmospheric emissions of Hg, As, and Se from coal fired power plants in China are evaluated. The national total emissions of Hg, As, and Se from coal-fired power plants in 2007 are calculated at 132 t, 550 t, and 787 t, respectively. Furthermore, according to the percentage of coal consumed by units equipped with different types of PM devices and FGD systems, speciation of mercury is estimated as follows: 80.48 t of Hg, 49.98 t of Hg(2+), and 1.89 t of Hg(P), representing 60.81%, 37.76%, and 1.43% of the totals, respectively. The emissions of Hg, As, and Se in China's eastern and central provinces are much higher than those in the west, except for provinces involved in the program of electricity transmission from west to east China, such as Sichuan, Guizhou, Yunnan, Shaanxi, etc. PMID- 21621817 TI - Biochar for the mitigation of nitrate leaching from soil amended with biosolids. AB - Countries with sewage treatment plants produce on average 27 kg of dried biosolids/person/yr. Concerns about nitrate leaching limit the rate at which biosolids are added to soil. We sought to determine whether biochar, a form of charcoal that is added to soil, could reduce nitrate leaching from biosolids amended soil. We set up 24 (0.5 m * 0.75 m) lysimeters, filled with two soil types (Templeton Silt Loam and Ashley Dene silt loam) and amended with combinations of biochar (102 t/ha equivalent) and biosolids (600 and 1200 kg N/ha equivalent). Pasture and leachates were sampled over 5 months. Nitrate leaching from biochar plus biosolids amended soils were reduced to levels at or below the control treatments. Pasture N concentrations were similarly affected by biochar addition. Future research should focus on unravelling the mechanism responsible for the change in the nitrogen cycle in soils amended with biosolids and biochar. PMID- 21621818 TI - Efficacy of using multiple open-path Fourier transform infrared (OP-FTIR) spectrometers in an odor emission episode investigation at a semiconductor manufacturing plant. AB - This study evaluated the efficacy of simultaneously employing three open-path Fourier transform infrared (OP-FTIR) spectrometers with 3-day consecutive monitoring, using an odor episode as an example. The corresponding monitoring paths were allocated among the possible emission sources of a semiconductor manufacturing plant and the surrounding optoelectronic and electronic-related factories, which were located in a high-tech industrial park. There was a combined total odor rate of 43.9% for the three monitoring paths, each comprised of 736 continuous 5-minute monitoring records and containing detectable odor compounds, such as ammonia, ozone, butyl acetate, and propylene glycol monomethyl ether acetate (PGMEA). The results of the logistic regression model indicated that the prevailing south wind and the OP-FTIR monitoring path closest to the emission source in down-wind direction resulted in a high efficacy for detecting odorous samples with odds ratios (OR) of 3.8 (95% confidence interval (CI): 2.9 5.0) and 5.1 (95% CI: 3.6-7.2), respectively. Meanwhile, the odds ratio for detecting ammonia odorous samples was 7.5 for Path II, which was downwind closer to the possible source, as compared to Path III, downwind far away from the possible source. PGMEA could not be monitored at Path II but could be at Path III, indicating the importance of the monitoring path and flow ejection velocities inside the stacks on the monitoring performance of OP-FTIR. Besides, an odds ratio of 5.1 for odorous sample detection was obtained with south prevailing wind comprising 65.0% of the monitoring time period. In general, it is concluded that OP-FTIR operated with multiple paths simultaneously shall be considered for investigation on relatively complicated episodes such as emergency of chemical release, multiple-source emission and chemical monitoring for odor in a densely populated plant area to enhance the efficacy of OP-FTIR monitoring. PMID- 21621819 TI - DNA damage in Gammarus fossarum sperm as a biomarker of genotoxic pressure: intrinsic variability and reference level. AB - In the perspective of a biomonitoring application for assessing genotoxicity of freshwater ecosystems, the Comet assay has recently been developed on spermatozoa in the amphipod Gammarus fossarum, in order to propose a sensitive and reliable genotoxicity biomarker in an ecologically relevant freshwater species. The appropriate use of a genotoxicity biomarker requires good knowledge of its basal level and its natural variability related to intrinsic biotic and environmental abiotic factors. We propose a procedure for which the lowest biomarker variability related to methodological and intrinsic biotic factors is obtained and a reference value of biomarker basal response taking into account its spatio temporal changes has been defined. A strong impact of spermatogenesis status and exposure time on the response to genotoxicant pressure was observed. These reports led us to select a standard organism, i.e., the mature male gammarid in precopula. No effect of temperature and conductivity on baseline DNA damage was observed in the laboratory for the tested range (6-24 degrees C and 300/600 MUS cm-1). Similarly, no spatio-temporal change relative to season or the physico chemical characteristics of the water was recorded during the field survey. On the basis of these results, a reference level with maximal threshold values has been proposed for the standard gammarid. PMID- 21621820 TI - Resistance and recovery of river biofilms receiving short pulses of Triclosan and Diuron. AB - The effects of the herbicide Diuron (DIU) and the bactericide Triclosan (TCS) were assessed on laboratory-grown stream biofilms. Four week-old biofilms were exposed in mesocosms to 48-hours of short pulses of either DIU or TCS. The direct and indirect effects of each toxicant on the biofilms, and the subsequent recovery of the biofilms, were evaluated according to structural and functional biomarkers. These parameters were analyzed immediately before exposure, immediately after exposure, and 9 and 16days post-exposure. DIU caused an increase in diatom mortality (+79%), which persisted until the end of the experiment. TCS also affected diatom mortality (+41%), although the effect did not appear until 1week post-exposure. TCS caused an increase in bacterial mortality (+45%); however, this parameter returned to normal values 1week post exposure. TCS compromised the cellular integrity of the green alga Spirogyra sp., whereas DIU did not. TCS also strongly inhibited phosphate uptake (-71%), which did not return to normal values until 2weeks post-exposure. DIU directly affected algae, but barely affected the heterotrophs, whereas TCS seriously impaired bacteria (direct effect) as well as autotrophs (indirect effect). However, the biofilms recovered their normal structure and function within only a few days to a few weeks. These findings demonstrate the capacity of biofilms to cope with periodic inputs of toxicants, but also the risks associated to repeated exposure or multi-contamination in aquatic ecosystems. PMID- 21621821 TI - Phytotoxicity testing for herbicide regulation: shortcomings in relation to biodiversity and ecosystem services in agrarian systems. AB - The purpose of this paper is to present current knowledge on methods employed to perform phytotoxicity tests and risk assessments and to highlight shortcomings in relation to biodiversity and ecosystem services. Ecosystem services are benefits provided to humankind by a multitude of organisms present in natural ecosystems. Several studies were conducted between 2001 and 2010 aimed at investigating some of the deficiencies in phytotoxicity testing (new and existing data are presented). Herbicide toxicity responses were similar when comparing a suite of crop versus wild species. However, the validity of the evaluation was limited because of the narrow types of species tested. The number of species tested, currently set between six and ten, appears insufficient. The trait-based approach (i.e. the use of plant attributes to predict species sensitivity to toxicants) can be used to improve species selection. This approach puts more emphasis on the shared biological characteristics that affect plant function within ecological communities rather than on plant phylogeny. Results presented showed that further studies are needed. In test guidelines, protocols require that crop species be sprayed as young vegetative plants, which is assumed to be the most sensitive growth stage to herbicides. In contrast, during herbicide spray, herbicides may reach non-target plants that are at various phenological stages. Several studies demonstrated that plants may be at greater risk when contamination occurs at the reproductive stage. No data on long-term effects, plant recovery or on effects on reproductive stages are requested in current guidelines. Preliminary evidence suggests that this may be an important aspect to consider in risk assessment. In addition, herbicide impacts on plant community diversity as well as biodiversity at other trophic levels have been demonstrated in only a limited number of studies and therefore should warrant more attention in risk assessment. PMID- 21621822 TI - Migration behavior of landfill leachate contaminants through alternative composite liners. AB - Four identical pilot-scale landfill reactors with different alternative composite liners were simultaneously operated for a period of about 540 days to investigate and to simulate the migration behaviors of phenolic compounds (phenol, 2-CP, 2 MP, 3-MP, 4-MP, 2-NP, 4-NP, 2,4-DNP, 2,4-DCP, 2,6-DCP, 2,4,5-TCP, 2,4,6-TCP, 2,3,4,6-TeCP, PCP) and heavy metals (Pb, Cu, Zn, Cr, Cd, Ni) from landfill leachate to the groundwater. Alternative landfill liners of four reactors consist of R1: Compacted clay liner (10 cm+10 cm, k=10(-8)m/sn), R2: Geomembrane (2 mm HDPE)+compacted clay liner (10 cm+10 cm, k=10-8 m/sn), R3: Geomembrane (2 mm HDPE)+compacted clay liner (10 cm, k=10-8 m/sn)+bentonite liner (2 cm)+compacted clay liner (10 cm, k=10-8 m/sn), and R4: Geomembrane (2 mm HDPE)+compacted clay liner (10 cm, k=10-8 m/sn)+zeolite liner (2 cm)+compacted clay liner (10 cm, k=10 8 m/sn). Wastes representing Istanbul municipal solid wastes were disposed in the reactors. To represent bioreactor landfills, reactors were operated by leachate recirculation. To monitor and control anaerobic degradation in the reactors, variations of conventional parameters (pH, alkalinity, chloride, conductivity, COD, TOC, TKN, ammonia and alcaly metals) were also investigated in landfill leachate samples. The results of this study showed that about 35-50% of migration of organic contaminants (phenolic compounds) and 55-100% of migration of inorganic contaminants (heavy metals) to the model groundwater could be effectively reduced with the use of bentonite and zeolite materials in landfill liner systems. Although leachate contaminants can reach to the groundwater in trace concentrations, findings of this study concluded that the release of these compounds from landfill leachate to the groundwater may potentially be of an important environmental concern based on the experimental findings. PMID- 21621823 TI - Emission characteristics of a heavy-duty diesel engine at simulated high altitudes. AB - In order to evaluate the effects of altitude on the pollutant emissions of a diesel engine, an experimental research was carried out using an engine test bench with an altitude simulation system. The emissions of HC, CO, NOx, smoke, and particle number of a heavy-duty diesel engine were measured under steady state operating conditions at sea level and simulated altitudes of 1000 and 2000 m. The experimental results indicate that the high altitude increases the emissions of HC, CO and smoke of the diesel engine, the average increasing rates of which are 30%, 35% and 34% with addition of altitude of 1000 m, respectively. The effect of high altitudes on the NOx emission varies with the engine types and working conditions. At 1000 m the particles number emissions are 1.6 to 4.2 times the levels at the low altitude. The pattern of the particle size distributions at 1000 m is similar with that at sea-level, which is the mono-modal lognormal distribution with geometric mean diameter around 0.1 MUm. However, the peak number concentrations of particles are bigger and the exhausted particles are smaller at the high altitude. PMID- 21621824 TI - Pharmacokinetics, distribution, and excretion of 125I-labeled human plasma derived-FVIIa and -FX with MC710 (FVIIa/FX mixture) in rats. AB - INTRODUCTION: MC710 is a mixture agent consisting of plasma-derived activated factor VII (FVIIa) and factor X (FX) at a weight ratio of 1:10 developed as a novel bypassing agent for the management of the bleeding of hemophilia patients with inhibitors. The pharmacokinetics, distribution, and excretion of (125)I labeled-FVIIa ((125)I-FVIIa) and -FX ((125)I-FX) were studied in male rats after a single intravenous administration of (125)I-FVIIa or (125)I-FX combined with MC710. METHODS: (125)I-FVIIa or (125)I-FX was administered intravenously with MC710 to male rats in a single dosage (FVIIa 0.4 mg and FX 4 mg/kg body weight) and radioactivity and antigen levels in plasma were quantified for 24h. Urine and feces were sampled to study the excretion of radioactivity during 168 h after dosing. Whole-body autoradiography was performed to evaluate the qualitative distribution of radioactivity 168 h after dosing. RESULTS AND CONCLUSIONS: The half-life (t(1/2)alpha and t(1/2)beta) of radioactivity and FVIIa antigen were 0.704 and 6.27 h, and 0.496 and 1.66 h, respectively and the area under the plasma concentration-time curve (AUC(0-infinity)) of radioactivity and FVIIa antigen were 17,932 and 8671 ng.h/mL, respectively. The t(1/2) of radioactivity and FX antigen were 4.06 and 3.05 h, respectively, and the AUC(0-infinity) of radioactivity and FX antigen were 320,143 and 395,794 ng.h/mL, respectively. About 80% of the administered dose of radioactivity was excreted in urine and feces by 168 h after administration. Tissue distribution experiments showed that FVIIa- and FX-related (125)I accumulated in bone and bone marrow, and disappeared slowly. PMID- 21621825 TI - Upregulation of hypoxia-inducible factor 1 alpha in local vein wall is associated with enhanced venous thrombus resolution. AB - INTRODUCTION: Venous thrombus resolution may be regulated by an angiogenic process that involves the surrounding vein wall. The aims of this study were to determine whether: (i) thrombosis stimulates activation of the angiogenic transcription factor, hypoxia-inducible factor (HIF) 1alpha, and downstream expression of growth factors in vein wall; and (ii) upregulation of HIF1alpha in vein wall leads to increased growth factor expression and enhanced thrombus resolution. MATERIALS AND METHODS: HIF1alpha, vascular endothelial growth factor (VEGF), and placental growth factor (PLGF) were quantified in mouse inferior vena cava (IVC) at days 1, 3, 7, and 14 after thrombus formation (n = 10-13 per group). An additional group of thrombosed mice were treated with the prolyl hydroxylase domain (PHD) inhibitor, L-mimosine (L-mim) or vehicle control. HIF1alpha, VEGF, and PLGF in IVC were measured at days 1 and 7; and vein recanalisation and thrombus resolution were measured at days 7 and 10 (n = 6-7 per group). RESULTS: HIF1alpha was expressed in thrombosed IVC and its levels remained relatively constant throughout natural resolution. The levels of VEGF in thrombosed IVC were elevated at days 1 (P < 0.0001) and 3 (P < 0.05); and PLGF at days 1 (P < 0.0001), 3 (P < 0.0001), and 7 (P < 0.0001). Treatment with L-mim led to: increased HIF1alpha (P<0.05), VEGF (P < 0.005), and PLGF (P < 0.001) levels in the IVC; decreased thrombus size (P < 0.01); and increased vein recanalisation (P < 0.001). CONCLUSIONS: HIF1alpha levels in vein wall are not affected by thrombosis and it appears that the angiogenic drive in the vein surrounding resolving thrombus is regulated independently of HIF1alpha. Stimulating HIF1alpha levels in the vein wall leads to an increased angiogenic drive and promotes vein recanalisation and thrombus resolution. PMID- 21621826 TI - Acquired factor VIII inhibitor as presentation of chronic myelogenous leukemia during interferon-alpha therapy. PMID- 21621827 TI - The expression and role of c-Myc in mouse hair follicle morphogenesis and cycling. AB - Although the function of c-Myc has been clarified in many tissues, until now its expression and role in hair follicle morphogenesis and the hair cycle remains unknown. In this study we detected c-Myc expression pattern in the process of mouse hair follicle development and normal cycle. We found that during hair follicle morphogenesis, the stage-specific expression of c-Myc was detected in mouse skin and was predominantly localized to the hair follicle epithelium. c-Myc expression was also consistently found in mouse skin throughout the hair follicle cycle. Through the in vivo injection of c-Myc inhibitory peptide and c-Myc expression plasmid, we also investigated the direct effects of c-Myc on the hair follicle structures during the hair follicle cycle. Our results showed that c-Myc inhibitory peptide significantly restrained the development of anagen hair follicles, while the injection of plasmid DNA encoding c-Myc in vivo clearly promoted anagen development. Our data indicate that c-Myc may play an important role in the proliferation and differentiation of the hair follicle keratinocytes during hair follicle development. c-Myc also was shown to participate in the regulation of the mouse hair growth cycle and could promote the proliferation of the hair matrix keratinocytes as well as the differentiation of the inner root sheath. PMID- 21621828 TI - In situ hybridization and immunohistochemical localization of leptin hormone and leptin receptor in the seminal vesicle and prostate gland of adult rat. AB - The role of leptin in the regulation of male reproductive function is still a matter of debate. Knowledge about a possible source of leptin in the seminal plasma may therefore be helpful in identifying and elucidating the physiological role of leptin hormone in male reproduction. In our investigation, the expression of leptin and its long receptor isoform (Ob-Rb) was studied in adult male Wistar rats using RT-PCR, Southern blot, in situ hybridization and immunohistochemistry. RT-PCR analysis revealed the expression of both leptin and its Ob-Rb in the seminal vesicle and prostate gland. In situ hybridization also localized the mRNA transcripts of leptin and Ob-Rb in the glandular secretory epithelial cells of prostate gland and seminal vesicle. Immunohistochemistry detected the leptin hormone in the lining epithelium of both male genital glands. In conclusion, these findings suggest that the seminal vesicle and prostate gland could be the possible sources of leptin in the seminal plasma. This leptin might have a direct (paracrine, autocrine or both) effect on epithelial cells of the accessory male genital glands, on the spermatozoa via spermatozoan leptin receptors. PMID- 21621829 TI - Ontogenetic anatomy of the distal vagina: relevance for local tumor spread and implications for cancer surgery. AB - OBJECTIVE: We have suggested to base cancer surgery on ontogenetic anatomy and the compartment theory of tumor permeation in order to improve local tumor control and to lower treatment-related morbidity. Following the validation of this concept for the uterine cervix, proximal vagina and vulva, this study explores its applicability for the distal vagina. METHODS: Serial transverse sections of female embryos and fetuses aged 8-17 weeks were assessed for the morphological changes in the region defined by the deep urogenital sinus-vaginal plate complex. Histopathological pattern analysis of local tumor spread was performed with carcinomas of the lower genital tract involving the distal vagina to test the compartment theory. RESULTS: Ontogenetically, the female urethra, urethrovaginal septum, distal vagina and rectovaginal septum represent a morphogenetic unit derived from the deep urogenital sinus-vaginal plate complex. Herein, the posterior urethra, the urethrovaginal septum and the distal vagina form a distinct subcompartment differentiated from the dorsal wall of the urogenital sinus. From 150 consecutive patients with distal vaginectomy as part of their surgical treatment 26 carcinomas of the lower genital tract had infiltrated the distal vagina. All 22 tumors involving the ventral wall invaded the urethra/periurethral tissue. Of the five carcinomas involving the dorsal wall none invaded the rectum/mesorectum. CONCLUSION: The pattern of local tumor permeation of lower genital tract cancer in the distal vagina can be consistently explained with ontogenetic anatomy and the compartment theory. PMID- 21621830 TI - Using biomarkers in the assessment of airways disease. AB - A biomarker provides a window on underlying disease activity. This is helpful when the pathology, treatment response, or both are heterogeneous or when trying to interpret nonspecific respiratory symptoms in patients with comorbidities. The successful application of a biomarker result is critically dependent on the specific question being addressed and the performance characteristics of the biomarker in relation to that question in the context of pretest probabilities. Negative prediction might be the best way to use a biomarker, such as a D-dimer, pro-brain natriuretic peptide, and exhaled nitric oxide. In this review the role of biomarkers in airways disease (notably induced sputum eosinophils and exhaled nitric oxide) is considered in relation to risk stratification, identification of treatment responders, identification of a clinical phenotype, monitoring of disease, and new drug development. PMID- 21621831 TI - Inhaled corticosteroid dosing: double for nothing? AB - Two recent trials from the National Heart, Lung, and Blood Institute's asthma clinical trials networks raise a concern about using double the dose of an inhaled corticosteroid (ICS) as a positive control arm in clinical trials of add on therapy. The literature evaluating the response to doubling the dose of an ICS is briefly reviewed. The vast majority of studies do not demonstrate a significant positive benefit from doubling the dose of an ICS but do show improvement with 4-fold increases that is equal to or greater than that of add-on long-acting bronchodilators. It is recommended that doubling the dose of an ICS no longer be considered a positive comparator arm in clinical trials, although it might be beneficial in individual patients. PMID- 21621832 TI - Nurse champions: a key role in bridging the gap between research and practice. PMID- 21621833 TI - Gene therapy for critical limb ischaemia: the TAMARIS trial. PMID- 21621834 TI - Effect of fibroblast growth factor NV1FGF on amputation and death: a randomised placebo-controlled trial of gene therapy in critical limb ischaemia. AB - BACKGROUND: Patients with critical limb ischaemia have a high rate of amputation and mortality. We tested the hypothesis that non-viral 1 fibroblast growth factor (NV1FGF) would improve amputation-free survival. METHODS: In this phase 3 trial (EFC6145/TAMARIS), 525 patients with critical limb ischaemia unsuitable for revascularisation were enrolled from 171 sites in 30 countries. All had ischaemic ulcer in legs or minor skin gangrene and met haemodynamic criteria (ankle pressure <70 mm Hg or a toe pressure <50 mm Hg, or both, or a transcutaneous oxygen pressure <30 mm Hg on the treated leg). Patients were randomly assigned to either NV1FGF at 0.2 mg/mL or matching placebo (visually identical) in a 1:1 ratio. Randomisation was done with a central interactive voice response system by block size 4 and was stratified by diabetes status and country. Investigators, patients, and study teams were masked to treatment. Patients received eight intramuscular injections of their assigned treatment in the index leg on days 1, 15, 29, and 43. The primary endpoint was time to major amputation or death at 1 year analysed by intention to treat with a log-rank test using a multivariate Cox proportional hazard model. This trial is registered with ClinicalTrials.gov, number NCT00566657. FINDINGS: 259 patients were assigned to NV1FGF and 266 to placebo. All 525 patients were analysed. The mean age was 70 years (range 50-92), 365 (70%) were men, 280 (53%) had diabetes, and 248 (47%) had a history of coronary artery disease. The primary endpoint or components of the primary did not differ between treatment groups, with major amputation or death in 86 patients (33%) in the placebo group, and 96 (36%) in the active group (hazard ratio 1.11, 95% CI 0.83-1.49; p=0.48). No significant safety issues were recorded. INTERPRETATION: TAMARIS provided no evidence that NV1FGF is effective in reduction of amputation or death in patients with critical limb ischaemia. Thus, this group of patients remains a major therapeutic challenge for the clinician. FUNDING: Sanofi-Aventis, Paris, France. PMID- 21621835 TI - Pro-osteogenic trophic effects by PKA activation in human mesenchymal stromal cells. AB - Human mesenchymal stromal cells (hMSCs) are able to differentiate into a wide variety of cell types, which makes them an interesting source for tissue engineering applications. On the other hand, these cells also secrete a broad panel of growth factors and cytokines that can exert trophic effects on surrounding tissues. In bone tissue engineering applications, the general assumption is that direct differentiation of hMSCs into osteoblasts accounts for newly observed bone formation in vivo. However, the secretion of bone-specific growth factors, but also pro-angiogenic factors, could also contribute to this process. We recently demonstrated that secretion of bone specific growth factors can be enhanced by treatment of hMSCs with the small molecule db-cAMP (cAMP) and here we investigate the biological activity of these secreted factors. We demonstrate that conditioned medium contains a variety of secreted growth factors, with differences between medium from basic-treated and cAMP-treated hMSCs. We show that conditioned medium from cAMP-treated hMSCs increases proliferation of various cell types and also induces osteogenic differentiation, whereas it has differential effects on migration. Microarray analysis on hMSCs exposed to conditioned medium confirmed upregulation of pathways involved in proliferation as well as osteogenic differentiation. Our data suggests that trophic factors secreted by hMSCs can be tuned for specific applications and that a good balance between differentiation on the one hand and secretion of bone trophic factors on the other, could potentially enhance bone formation for bone tissue engineering applications. PMID- 21621836 TI - Chemically amplified photoresist for high resolution autoradiography in targeted radiotherapy. AB - Evaluation of the intracellular distribution of radionuclides used for targeted radiotherapy (tRT) is essential for accurate dosimetry. Therefore, a direct and quantitative method for subcellular micro-autoradiography using radiation sensitive polymers (PMMA, UV1116 and AZ40XT) was developed. The electron exposure dose in radio-labelled cells due to Auger and internal conversion (IC) electron emissions of indium (111In), a radionuclide currently used for tRT, was calculated using Monte Carlo (MC) simulation. Electron beam lithography using pre defined exposure doses was used to calibrate the resist response. The topography of the exposed and developed resists was analysed with atomic force microscopy (AFM) and the resulting pattern depth was related to a specific exposure dose. UV1116 exhibited the best contrast as compared to AZ40XT and PMMA, while AZ40XT exhibited the highest sensitivity at low doses (<10 MUC/cm2). AFM analysis of the exposure pattern from radio-labelled cells and nuclei in UV1116 revealed a non uniform distribution of 111In-EGF in the cell and nucleus, consistent with less well-resolved data from confocal microscopy and micro-autoradiography. PMID- 21621837 TI - Three-dimensional porous silk tumor constructs in the approximation of in vivo osteosarcoma physiology. AB - The lack of good preclinical models has hampered anticancer drug discovery. Standard preclinical protocols require the growth of cells in high throughput two dimensional (2D) culture systems. However, such in vitro drug testing methods yield drug efficacy results that differ greatly from animal models. Conversely, it is much more difficult and expensive to use animal models for large-scale molecular biology research. It is conceivable that three-dimensional (3D) growth may be responsible for some of these changes. Porous silk sponges were fabricated through freeze drying and seeded with 143.98.2 osteosarcoma cells. Molecular profiles were obtained by carrying out real-time polymerase chain reaction for angiogenic growth factors and proliferation markers for osteosarcoma cells grown under 2D, 3D, and SCID mouse xenograft conditions. The angiogenic factor expression profiles for cells grown in 2D differed greatly from the 3D silk scaffold model (P < 0.05 for bFGF, HIF-1alpha, IL-8, and VEGF-A), whereas 3D tumor model profiles were found to be able to approximate that for the in vivo tumor better with no statistically different expression of HIF-1alpha and VEGF-A between the two. Immunohistochemistry staining for HIF-1alpha, VEGF-A, and VEGF receptor on osteosarcoma cells grown on the scaffolds validated the results obtained with the gene expression profiles. The results suggest that 3D tumor models could be used to bridge the gap between in vitro and in vivo tumor studies, and aid in the study of mechanisms activated during tumorigenesis for the development of novel targeted chemotherapy. PMID- 21621838 TI - The influence of substrate creep on mesenchymal stem cell behaviour and phenotype. AB - Human mesenchymal stem cells (hMSCs) are capable of probing and responding to the mechanical properties of their substrate. Although most biological and synthetic matrices are viscoelastic materials, previous studies have primarily focused upon substrate compressive modulus (rigidity), neglecting the relative contributions that the storage (elastic) and loss (viscous) moduli make to the summed compressive modulus. In this study we aimed to isolate and identify the effects of the viscous component of a substrate on hMSC behaviour. Using a polyacrlyamide gel system with constant compressive modulus and varying loss modulus we determined that changes to substrate loss modulus substantially affected hMSC morphology, proliferation and differentiation potential. In addition, we showed that the effect of substrate loss modulus on hMSC behaviour is due to a reduction in both passive and actively generated isometric cytoskeletal tension caused by the inherent creep of substrates with a high loss modulus. These findings highlight substrate creep, or more explicitly substrate loss modulus, as an important mechanical property of a biomaterial system that can be tailored to encourage the growth and differentiation of specific cell types. PMID- 21621839 TI - Stereology of the placenta in type 1 and type 2 diabetes. AB - OBJECTIVE: To assess by stereology the placental structure in type 1 (T1DM) and type 2 (T2DM) diabetic pregnancies compared to normal non-diabetic (ND) controls. STUDY DESIGN: Prospective case control study. Placentae were sampled in a systematic random fashion. Stereological analysis was performed using a computerised stereology programme (Image Pro 6.2, Media Cybernetics, Inc, Silver Spring MD, USA). Participants were matched for gender of infant and mode of delivery. MAIN OUTCOME MEASURES: Volume, length and surface area of placental components; clinical outcome. RESULTS: Ten ND, eight T2DM and ten T1DM women consented to the study. There was no difference between the groups regarding maternal age, neonatal birth weight, or placental weight. On stereological examination, terminal villous volume was significantly increased in both diabetic groups compared to ND controls. Capillary volume and length was increased in T1DM pregnancies compared to ND and T2DM. Capillary length was increased in both diabetic groups compared to ND. When all diabetic groups were compared based on severity of glycaemia those with poor glycaemic control (HbA1c>7%) had higher placental capillary volume than those with good glycaemic control. CONCLUSIONS: This study demonstrates an association between maternal diabetes and increased terminal villous volume. Additionally capillary volume and length is increased in the placentae of normally grown infants of T1DM diabetic mothers compared to non diabetic controls. Maternal glycaemia appears to influence capillary, but not stromal, development. This suggests that factors other than glycaemia have a role in placental development in pre-gestational diabetes. PMID- 21621840 TI - Calcium and connexin-based intercellular communication, a deadly catch? AB - Ca(2+) is known as a universal messenger mediating a wide variety of cellular processes, including cell death. In fact, this ion has been proposed as the 'cell death master', not only at the intracellular but also at the intercellular level. The most direct form of intercellular spread of cell death is mediated by gap junction channels. These channels have been shown to propagate cell death as well as cell survival signals between the cytoplasm of neighbouring cells, reflecting the dual role of Ca(2+) signals, i.e. cell death versus survival. Its precursor, the unopposed hemichannel (half of a gap junction channel), has recently joined in as a toxic pore connecting the intracellular with the extracellular environment and allowing the passage of a range of substances. The biochemical nature of the so-called intercellular cell death molecule, transferred through gap junctions or released/taken up via hemichannels, remains elusive but several studies pinpoint Ca(2+) itself or its messenger inositol trisphosphate as the responsible masters in crime. Although direct evidence is still lacking, indirect data including Ca(2+) involvement in intercellular communication and cell death, and effects of intercellular communication on intracellular Ca(2+) homeostasis, support this hypothesis. In addition, hemichannels and their molecular building blocks, connexin or pannexin proteins, may exert their effects on Ca(2+) dependent cell death at the intracellular level, independently from their channel functions. This review provides a cutting edge overview of the current knowledge and underscores the intimate connection between intercellular communication, Ca(2+) signalling and cell death. PMID- 21621841 TI - Effects of pituitary adenylate cyclase activating polypeptide in the urinary system, with special emphasis on its protective effects in the kidney. AB - Pituitary adenylate cyclase activating polypeptide (PACAP) is a widespread neuropeptide with diverse effects in the nervous system and peripheral organs. One of the most well-studied effects of PACAP is its cytoprotective action, against different harmful stimuli in a wide variety of cells and tissues. PACAP occurs in the urinary system, from the kidney to the lower urinary tract. The present review focuses on the nephroprotective effects of PACAP and summarizes data obtained regarding the protective effects of PACAP in different models of kidney pathologies. In vitro data show that PACAP protects tubular cells against oxidative stress, myeloma light chain, cisplatin, cyclosporine-A and hypoxia. In vivo data provide evidence for its protective effects in ischemia/reperfusion, cisplatin, cyclosporine-A, myeloma kidney injury, diabetic nephropathy and gentamicin-induced kidney damage. Results accumulated on the renoprotective effects of PACAP suggest that PACAP is an emerging candidate for treatment of human kidney pathologies. PMID- 21621842 TI - Amount of bone marrow blasts is strongly correlated to NPM1 and FLT3-ITD mutation rate in AML with normal karyotype. AB - FLT3-ITDs are linked to higher leukocytes/blasts in acute myeloid leukemia. To evaluate the effect of NPM1 mutations, we correlated NPM1mut status with morphology in 805 adult normal karyotype AML. NPM1mut were found in 391/805 (48.6%), FLT3-ITD in 219/805 (27.2%). Frequencies of FLT3-ITD and NPM1mut cases were continuously increasing by blast decades: NPM1mut from 38/123 (30.9%) in 20 29% blast decade to 70/103 (68.0%) in 90-100% decade (p<0.001), FLT3-ITDs from 15/123 (12.2%) to 58/103 (56.3%) (p<0.001). Mean WBC count was highest in NPM1 mut/FLT3-ITD-positive and lowest in NPM1-wildtype/FLT3-ITD-negative patients (p<0.0001); similar for BM blasts. Therefore, FLT3-ITD and NPM1mut might synergistically stimulate blast proliferation. PMID- 21621843 TI - Examining the effects of metal mining mixtures on fathead minnow (Pimephales promelas) using field-based multi-trophic artificial streams. AB - This study illustrates the use of a mesocosm approach for assessing the independent effects of three treated metal mine effluents (MME) discharging into a common receiving environment and regulated under the same regulation. A field based, multi-trophic artificial stream study was conducted in August 2008 to assess the effects of three metal mining effluents on fathead minnow (Pimephales promelas) in a 21-day reproduction bioassay. The nature of the approach allowed for assessment of both dietary and waterborne exposure pathways. Elements (e.g. Se, Co, Cl, Cu, Fe) were analyzed in several media (water, sediments) and tissues (biofilm, Chironomus dilutus, female fathead minnow (FHM) body, ovary, liver, gills). Significant increases in metal and micronutrient concentrations were observed in the water and biofilm tissues in all MME treatments [20% surface water effluent (SWE), 30% mine water effluent (MWE), and 45% process water effluent (PWE)], compared to reference. However, copper was the only element to significantly increase in the sediments when exposed to PWE. Co and Ni increased significantly in C. dilutus tissues in SWE (1.4- and 1.5-fold, respectively), Cu and Se also increased in chironomid tissues in PWE (5.2- and 3.3-fold, respectively); however, no significant increases in metals or micronutrients occurred in chironomid tissues when exposed to MWE compared to reference. There were no significant increases in metal concentrations in female FHM tissues (body, liver, gonads, gills) in any of the treatments suggesting that metals were either not bioavailable, lost from the females via the eggs, or naturally regulated through homeostatic mechanisms. Cumulative number of eggs per female per day increased significantly (~127%) after exposure to SWE and decreased significantly (~33%) after exposure to PWE when compared to reference. Mean total number of days to hatch was reduced in PWE compared to reference. This study shows the importance of isolating treatment streams in cumulative discharge environments to assess aquatic effects due to the different nature of the effluents. PMID- 21621844 TI - Sources and environmental behavior of dechlorane plus--a review. AB - Intensive research has been carried out in the past few years on the occurrence and behavior of dechlorane plus (DP) in the environment, biota and humans. This review summarizes the information and research results generated since DP was first identified in the environment in 2006. We describe the reported measurement methods for DP and provide an integrated picture of its occurrence and behavior as an environmental contaminant. DP in ambient air and sediments is characterized by strong source related concentration elevations and temporal trends reflecting commercial use. Long-range atmospheric transportation of DP has been observed in remote regions, indicating a global presence of DP. Levels of DP in biota and in humans further reinforce its source related distribution. The change in DP isomer ratio in various environmental matrices from commercial products indicates that the behavior of the two isomers is not the same in the environment or in biota. With only limited data available so far, the biomagnification of DP in aquatic biota has not been conclusively demonstrated. This review also looks at reported dechlorination products of DP that might be of interest in future research. PMID- 21621845 TI - Differential regulation of Nr4a subfamily nuclear receptors following mast cell activation. AB - The biological function of the Nr4a subfamily of nuclear receptors is only partially understood. Here we show for the fist time that mast cell (MC) activation processes involve the regulation of Nr4a factors. Exposure of murine bone marrow-derived MCs (BMMCs) to live bacteria causes a robust and selective upregulation of all Nr4a members (Nr4a1-Nr4a3). In response to purified LPS, strong upregulation of Nr4a3, but not of Nr4a1 or Nr4a2 was seen. Nr4a3 expression was also induced after the activation of BMMCs by IgE receptor cross linking. Moreover, Nr4a expression was induced in activated human MCs. As shown by Western blot analysis, Nr4a phosphorylation was induced by IgE receptor cross linking and calcium ionophore stimulation of BMMCs and LAD2 cells, respectively. By using various inhibitors of signaling pathways, Nr4a3 induction in BMMCs was shown to be strongly dependent on Go6976-sensitive kinases and partially dependent on the nuclear factor of activated T-cells (NFAT) pathway, while nuclear factor kappa-light-chain-enhancer of activated B cells (NFkappaB) inhibition failed to inhibit Nr4a3 expression in BMMCs. Together, these data reveal selective induction of Nr4a family members in activated MCs and implicate Nr4a family nuclear receptors in the regulation of MC function. PMID- 21621846 TI - Activation of mitogen activated protein kinase-Erk kinase (MEK) increases T cell immunoglobulin mucin domain-3 (TIM-3) transcription in human T lymphocytes and a human mast cell line. AB - The immune regulatory molecule T cell immunoglobulin mucin domain (TIM-3) is expressed in activated T cells and in mast cells treated with transforming growth factor (TGF)-beta, but underlying mechanisms for induction of TIM-3 transcription have not been well-explored. We studied the role of mitogen-activated protein kinase (MAPK) in TIM-3 transcription on the basis of the involvement of MAPK in T cell activation and TGF-beta signaling. Inhibitors of MAPK-Erk kinase (MEK) as well as p38 suppressed TIM-3 transcription in phorbol myristic acid (PMA) stimulated T cells, but inhibitors of c-Jun NH2-terminal kinase (JNK) did not. MEK over-expression enhanced TIM-3 transcription in PMA-stimulated T cells. Furthermore, -1.5kb TIM-3 promoter was activated by PMA stimulation and repressed by MEK inhibitors in Jurkat T cells. Similarly, MEK activation enhanced TIM-3 transcription in TGF-beta-stimulated HMC-1 human mast cells, although MEK seemed not directly activated by TGF-beta. Concordantly, -1.5kb TIM-3 promoter activity was reduced by MEK inhibitors, but was not responsive to TGF-beta stimulation in HMC-1 cells. These results suggest the regulatory role of MEK in TIM-3 transcription by human CD4+ T cells and mast cells. PMID- 21621847 TI - Physicians fail to consider Pseudomonas aeruginosa as a potential pathogen in medicine patients with bacteremia. PMID- 21621848 TI - Late presentation of HIV disease and its associated factors among newly diagnosed patients before and after abolition of a government policy of mass mandatory screening. AB - OBJECTIVE: To investigate the risk factors for late presentation in the Republic of Korea, where massive mandatory screening for HIV infection was conducted by the government until the late 1990s. METHODS: Data over the period 1987-2008 were analyzed from HIV patients for whom records of CD4 cell counts within 3 months of HIV diagnosis were available. Using multivariate logistic regression analysis including demographic and clinical variables, we examined factors associated with late presentation, defined as having a CD4 cell count of less than 200 cells/mm(3) at the time of diagnosis. RESULTS: Of a total of 994 patients with a new diagnosis of HIV infection, 405 (41%) were late presenters. As the proportion of patients diagnosed by mandatory screening decreased over time (31% in 1987 1998 versus 8% in 1999-2008, P < 0.001), the proportion of late presenters increased (31% in 1987-1998 versus 43% in 1999-2008, P = 0.007). The independent risk factors for late presentation were older age (adjusted odds ratio [aOR], per increase of 10 years, 1.31; 95% confidence interval [CI], 1.15-1.49; P < 0.001), male sex (aOR, 1.74; 95% CI, 1.03-2.95; P = 0.040), negativity for VDRL (aOR, 1.58; 95% CI, 1.16-2.14; P = 0.003), and diagnosis after 1999 (aOR, 1.64; 95% CI, 1.05-2.56; P = 0.031). CONCLUSIONS: Older age, male sex, negativity for VDRL, and diagnosis after 1999, were associated with late presentation, and the proportion of late presenters increased after the mandatory testing policy was abolished. PMID- 21621849 TI - Complicated grief among individuals with major depression: prevalence, comorbidity, and associated features. AB - BACKGROUND: Growing data suggest that complicated grief (CG) may be common in clinical care settings, but there are few prior reports about CG in outpatients presenting with primary mood disorders. METHODS: The present study examined rates of bereavement and threshold CG symptoms (defined as a score >= 25 on the Inventory of Complicated Grief scale) in 111 outpatients with major depressive disorder (MDD) and 142 healthy controls participating in a study of stress and depression. Clinical and demographic characteristics were also compared for bereaved individuals with CG (MDD+CG) to those without (MDD-CG). Participants completed structured diagnostic interviews as well as measures of CG, depression, anxiety, exposure to traumatic events, and perceived social support. RESULTS: Lifetime history of a significant loss did not differ for the MDD and control groups (79.3% vs. 76.1%), but bereaved participants with MDD had higher rates of threshold CG (25.0% vs. 2.8%). Among those with MDD, CG was associated with a higher prevalence of lifetime alcohol dependence, greater exposure to traumatic events, and lower perceived social support. Depressed women, but not men, with CG also had higher rates of panic disorder, social anxiety disorder, and posttraumatic stress disorder. LIMITATIONS: Our findings are limited by the lack of a clinician confirmatory assessment of CG diagnosis, absence of complete information about the nature and timing of the loss, and relatively narrow generalizability. CONCLUSIONS: We found high rates of CG in a group of psychiatric outpatients with chronic MDD, suggesting that patients with depression should be routinely screened for CG. PMID- 21621850 TI - Resilience mitigates the suicide risk associated with childhood trauma. AB - OBJECTIVE: We wished to examine whether resilience might be a protective factor in relation to suicidal behavior. METHOD: To do this resilience was examined in relation to childhood trauma, a well established risk factor for suicidal behavior, in two samples. In a preliminary sample 20 abstinent substance abuse patients who had attempted suicide were matched for age and their score on the Childhood Trauma Questionnaire (CTQ) with 20 substance abuse patients who had never attempted suicide. The two age and CTQ matched attempter (N=20) and non attempter (N=20) groups were then compared for their scores on the Connor Davidson Resilience Scale (CD-RISC). In the second sample 166 prisoners who had attempted suicide were matched for age and their scores on the CTQ with 166 prisoners who had never attempted suicide. These two age and CTQ matched attempter (N=166)and non-attempter (N=166) groups were similarly compared for their CD-RISC resilience scores. RESULTS: In the preliminary substance abuse sample, patients who had never attempted suicide (N=20) had significantly higher mean CD-RISC resilience scores than the age and CTQ matched patients who had attempted suicide (N=20). Similarly in the prisoner sample, those who had never attempted suicide (N=166) had significantly higher CD-RISC resilience scores than the age and CTQ matched prisoners who had attempted suicide (N=166). CONCLUSIONS: The results from these two studies suggest that resilience may be a protective factor mitigating the risk of suicidal behavior associated with childhood trauma. PMID- 21621851 TI - Characteristics of Norwegian children suffering from ADHD symptoms: ADHD and primary health care. AB - This study describes the characteristics of children suffering from attention deficit/hyperactivity disorder (ADHD) symptoms and evaluates the ability of primary health care to assess ADHD. A population of 494 children was referred during one year to the outpatient clinic Child and Adolescent Mental Health in Norway. Forty percent of those referred had ADHD symptoms. A clinical assessment regarding ADHD as well as general health was employed and socio-economic status was recorded. Half of the ADHD-referred children met criteria for ADHD, and among the non-ADHD children one fifth did not receive any diagnosis. Mean referral age was 10.5years; 82% were boys. The ADHD families were less educated and in more need of support from Child Welfare (CW) in the referral period (OR: 3.9; 95% CI 0.1 to 5.1). More ADHD children were not living with their families compared to the non-ADHD children. The sensitivity was 51% (96/187) regarding primary health care's ability to recognize ADHD. Further screening programs for evaluation of ADHD are needed. PMID- 21621852 TI - Life events and dementia: what is the nature of their relationship? AB - The current study analyzed the life events reported by 1271 demented patients vs. 140 cognitively healthy elderly subjects. The Life Change Unit (LCU) method was used to quantify the results. When all the events were included in the analysis, the two groups had similar LCU scores (61.26 vs. 63.42). However, when events causally related to dementia (e.g. stroke) are excluded, demented patients were found to experience half of the LCU load in comparison to controls (30.70 vs. 63.42). In both groups the level of LCU load is far below 100 which is the threshold suggested for the induction of psychosomatic disorders. Conclusively, the current study suggests that there is no causal role for life events in the etiopathogenesis of dementia. On the contrary, demented patients even the last few months before the clinical onset of dementia experience low life-events related stress, possibly because of subclinical impairment which is already present. PMID- 21621853 TI - Cigarette smoking in outpatients with chronic schizophrenia in Taiwan: relationships to socio-demographic and clinical characteristics. AB - In individuals with schizophrenia, the prevalence of cigarette smoking is significantly higher than that of the general population; this appears to be associated with specific psychosocial and clinical characteristics. Indeed, some evidence suggests an increased risk of suicide among smokers with schizophrenia. The purpose of this study was to examine the characteristics of smokers with schizophrenia in Taiwan. In this cross-sectional study, 95 outpatients with DSM IV diagnosis of schizophrenia were recruited and independently interviewed for nicotine dependency with tobacco use. The effects of cigarette smoking on the various measures, especially suicidality, were investigated. The results revealed that smokers with schizophrenia had higher rates of hospitalization, lifetime suicide attempts, antipsychotic treatment side effects, psychopathology, impulsivity, depression, anxiety, and suicidal risk than non-smokers with schizophrenia. When separate analyses were conducted in male and female patients, depressive symptoms were significant predictors of suicidality among males, whereas heavy smoking, anxious symptoms, and hopelessness were significant predictors among females. From this study, we may gain insights into the role of cigarette smoking in patients of schizophrenia in Taiwan. Furthermore, cigarette smoking may influence aspects of suicidality in schizophrenia. PMID- 21621854 TI - C-reactive protein serum level in drug-free male Egyptian patients with schizophrenia. AB - Despite the growing research interest in the role of immunological markers in schizophrenia, few studies, with conflicting results, have focused on the association between high sensitivity C-reactive protein (hs-CRP) levels and clinical characteristics in schizophrenia. In this cross-sectional case-control study, a sample of 200 antipsychotic-free male Egyptian schizophrenia patients was assessed by the Positive and Negative Syndrome Scale (PANSS) and compared with 200 healthy controls as regards serum hs-CRP level using an immunoturbidimetric method. CRP level for patients (geometric mean=3.3 mg/L) was significantly (P=0.000) higher than that for controls (geometric mean=1.4 mg/L). PANSS scores and patients' data, which significantly correlated with serum hs-CRP level, were entered into a stepwise multiple regression analysis. Results of this analysis showed that PANSS negative symptom score was second only to the waist circumference, with which they explained 54.7 % of the variation in serum hs-CRP. Comparable results were obtained when patients, controls and the relevant confounders were included in one multivariate analysis. We concluded that in Egyptian men, waist circumference and schizophrenia diagnosis are strong predictors of raised CRP level independent of a number of potentially confounding variables. In antipsychotic-free patients, CRP level is higher than in healthy controls and is positively correlated with the severity of the psychopathology as measured by PANSS. This relationship is especially notable in negative, but not positive symptoms. PMID- 21621855 TI - Single dose adenovirus vectored vaccine induces a potent and long-lasting immune response against rabbit hemorrhagic disease virus after parenteral or mucosal administration. AB - Rabbit hemorrhagic disease virus (RHDV) is the etiological agent of a lethal and contagious disease of rabbits that remains as a serious problem worldwide. As this virus does not replicate in cell culture systems, the capsid protein gene has been expressed in heterologous hosts or inserted in replication-competent viruses in order to obtain non-conventional RHDV vaccines. However, due to technological or safety issues, current RHDV vaccines are still prepared from organs of infected rabbits. In this work, two human type 5 derived replication defective adenoviruses encoding the rabbit hemorrhagic disease virus VP60 capsid protein were constructed. The recombinant protein was expressed as a multimer in mouse and rabbit cell lines at levels that ranged from approximately 120 to 160 mg/L of culture. Mice intravenously or subcutaneously inoculated with a single 10(8) gene transfer units (GTU) dose of the AdVP60 vector (designed for VP60 intracellular expression) seroconverted at days 7 and 14 post-immunization, respectively. This vector generated a stronger response than that obtained with a second vector (AdVP60sec) designed for VP60 secretion. Rabbits were then immunized by parenteral or mucosal routes with a single 10(9)GTU dose of the AdVP60 and the antibody response was evaluated using a competition ELISA specific for RHDV or RHDVa. Protective hemagglutination inhibition (HI) titers were also promptly detected and IgG antibodies corresponding with inhibition percentages over 85% persisted up to one year in all rabbits, independently of the immunization route employed. These levels were similar to those elicited with inactivated RHDV or with VP60 obtained from yeast or insect cells. IgA specific antibodies were only found in saliva of rabbits immunized by intranasal instillation. The feasibility of VP60 production and vaccination of rabbits with replication-defective adenoviral vectors was demonstrated. PMID- 21621856 TI - Increased serum matrix metalloproteinase-9 in neuromyelitis optica: implication of disruption of blood-brain barrier. AB - Matrix metalloproteinase-9 (MMP-9) plays an important role in some neuroinflammatory diseases through the blood-brain barrier (BBB) disruption. To investigate the pathogenicity of MMP-9 in neuromyelitis optica (NMO), serum and CSF MMP-9 concentrations were measured in 13 NMO and 15 multiple sclerosis (MS) patients and 14 healthy controls, and correlated with clinical and laboratorial parameters. Serum MMP-9 concentrations were significantly higher in NMO than MS and controls, and correlated with EDSS score, CSF/serum albumin ratio, and CSF IL 8 concentrations. Our results indicate that MMP-9, promoted by elevated IL-8 activation, plays a crucial role in the pathogenesis of NMO through the BBB disruption. PMID- 21621857 TI - Changes in responsiveness of appetite, leptin and hypothalamic IL-1beta and TNF alpha to lipopolysaccharide in developing rats. AB - In addition to its role as a regulator of energy homeostasis, leptin plays a pivotal role in some immune/inflammatory responses. Synthesis and secretion of leptin are increased under immune stress conditions, and increased leptin may participate in the development of anorexia and fever. These actions are partially mediated by up-regulation of hypothalamic IL-1beta. Leptin also protects against immune stress-induced lethality. On the other hand, the response and roles of leptin to immune stress conditions in the neonatal period have scarcely been examined. We hypothesized that 1. the response of leptin to immune stress would be suppressed in the early neonatal period, 2. hyporesponse of leptin in the early neonatal period would attenuate the anorectic response and increase the lethal rate under immune stress conditions and 3. supplementation of leptin in the early neonatal period would increase the anorectic response, whereas it would decrease the lethal rate under immune stress conditions. To test these hypotheses, we first examined the developmental changes in the responses of leptin and hypothalamic proinflammatory cytokines, i.e., IL-1beta and TNF-alpha, to LPS-induced immune stress in female rats. We also examined the developmental changes in the anorectic response and lethality rate under LPS-induced immune stress conditions. Five- and 15-day-old rats showed no leptin response and a weak hypothalamic IL-1beta response to LPS when compared with 25- and 42-day-old rats. Fifteen-day-old rats showed low anorectic responses and high lethality rates when compared with 25- and 42-day-old rats under LPS-induced immune stress conditions. We then examined whether administration of leptin affected the response to the anorectic and lethal effects of LPS in 15-day-old rats. Administration of leptin further attenuated body weight after LPS injection, but not after saline injection. On the other hand, administration of leptin did not affect survival rate. In addition, hypothalamic IL-1beta mRNA levels were not affected by leptin administration. In conclusion, the absence of a leptin response may act to prevent immune stress-induced anorexia during the early neonatal period. PMID- 21621858 TI - Anti-myelin associated glycoprotein antibodies recognize HNK-1 epitope on CNS. AB - Antibodies to myelin-associated glycoprotein (MAG) are associated with demyelinating polyneuropathy and are specific for the HNK-1 epitope. To test if anti-MAG IgM recognize HNK-1 on CNS, sera from 20 patients and 238 controls were tested on rat slices by indirect immunofluorescence (IIF). IgM from anti-MAG positive patients, but not from control sera, stained rat brain with perineuronal or neuropil pattern, depending on the CNS region. IIF titers significantly correlated with ELISA anti-MAG titers. The staining of patients' sera were inhibited by mouse anti-HNK-1 monoclonal antibody. Our results demonstrate that anti-MAG IgM recognizes HNK-1 outside the peripheral nerve myelin carriers. PMID- 21621859 TI - HLA DRB1*1501 is only modestly associated with lesion burden at the first demyelinating event. AB - OBJECTIVES: The presence of MRI lesions at the first demyelinating event increases the risk of developing clinically definite multiple sclerosis (MS). The HLA DRB1*1501 genotype is linked to MS susceptibility but its relationship to quantitative MRI parameters at the first demyelinating event has not been assessed. The objectives were to assess the associations between HLA DRB1*1501 status and magnetic resonance imaging (MRI) measures in clinically isolated syndromes (CIS) at the first demyelinating event. METHODS: We genotyped 205 CIS patients (age: 29.0+/-7.7 years) enrolled in the Observational Study of Early Interferon beta 1-a Treatment in High Risk Subjects after CIS (SET study), a multi-center, clinical study of CIS for rs3135005, a single nucleotide polymorphism associated with HLA DRB1*1501 status. The inclusion criteria required 2 or more brain MRI lesions and the presence of two or more oligoclonal bands in cerebrospinal fluid. Clinical and MRI assessments were obtained within 4 months of the initial demyelinating event. RESULTS: The frequency of HLA DRB1*1501 positivity was 102/205 (49.7%). HLA DRB1*1501 positivity was associated with higher contrast-enhancing (CE) lesion number (p=0.002), higher CE-lesion volume (LV) (p<0.001) and exhibited a trend with higher T2-LV (p=0.012). There was no evidence for significant associations of HLA DRB1*1501 positivity with disability, symptoms at CIS presentation, whole brain, white and gray matter atrophy. CONCLUSIONS: HLA DRB1*1501 positivity is associated with increased brain inflammatory processes at first clinical onset. However, the effect sizes of the HLA DRB1*1501 associations with MRI are modest, which potentially limits the clinical usefulness. PMID- 21621860 TI - Association of IL1A, IL1B, ILRN, IL6, IL10 and TNF-alpha polymorphisms with risk and clinical course of multiple sclerosis in a Polish population. AB - Single nucleotide polymorphisms in human pro- and anti-inflammatory genes, including IL1RN VNTR (rs315952), IL1A 4845G>T (rs17561), L1B-511C>T (rs16944), IL6-174G>C (rs1800795), IL10-1082 A>G (rs 1800896) and TNFalpha-308G>A (rs1800629) and their impact on multiple sclerosis risk and disease progression in a Polish population were investigated. Increased risk of MS was found for IL6 174 CC homozygotes (OR, 2.88; p<0.00001). In turn, IL1A 4845 TT genotype determined earlier appearance of MS onset whereas IL1B-511 TT genotype was associated with later occurrence of MS but faster disability progression. PMID- 21621861 TI - Endoscopic optic nerve decompression for traumatic optic neuropathy in children. AB - OBJECTIVES: To discuss the necessity of nasal endoscopic surgery for pediatric traumatic optic neuropathy and its therapeutic effect. METHODS: We retrospectively reviewed the cases of 41 children (involving 43 eyes) with traumatic optic neuropathies who were treated in our department by endoscopic optic nerve decompression and postoperative corticosteroid in large doses from Feb. 2000 to Apr. 2010. A 6-month follow-up study was performed for each patient in order to observe the postoperative eyesight recovery and analyze the therapeutic effect. RESULTS: The eyesight of 11 patients out of 41 patients reached 0.2-0.3 postoperatively, the eyesight of 16 patients recovered from counting fingers to 0.1 after the surgery, the eyesight of 6 patients ranged from light sensation to seeing the hand movement, the eyesight of 7 children did not recover from the operation. The prognosis in the children who underwent the decompression 7 days post-traumatically was much worse than other children. CONCLUSIONS: The age of the patients was not the main element influencing the decision-making process for the operation. The main elements affecting the prognosis were the degree of injury and the time interval between the trauma and the time when patients underwent the surgery. The operation opportunity and plan are very important to a successful operation. PMID- 21621862 TI - A clinical narrative review of mandibular distraction osteogenesis in neonates with Pierre Robin sequence. AB - INTRODUCTION: Neonatal upper airway obstruction secondary to micrognathia can be managed with conservative or surgical interventions. Traditionally, severe upper airway obstruction was managed with a tracheostomy. Although tracheostomy may be life saving, it is associated with high rates of complications and can lead to developmental problems. More recently, mandibular distraction osteogenesis has been utilized to relieve micrognathia associated airway obstruction. METHODS: A clinical narrative review of the current literature was performed to evaluate the efficacy of mandibular distraction osteogenesis in neonates with Pierre Robin sequence. OBJECTIVES: (1) To evaluate whether mandibular distraction osteogenesis can relieve the upper airway obstruction in micrognathic neonates and (2) to discuss and increase the awareness of various issues surrounding neonatal mandibular distraction procedures including preoperative workup, distraction protocols, and complications. RESULTS: Mandibular distraction osteogenesis can be a safe and effective intervention in neonates diagnosed with Pierre Robin sequence with severe micrognathia and airway obstruction. Interestingly, in patients with additional complex syndromes, the airway obstruction was not consistently alleviated. CONCLUSION: When conservative measures fail, mandibular distraction osteogenesis should be considered to obviate the need for a tracheostomy in newborns with micrognathia associated upper airway obstruction. PMID- 21621863 TI - Dynamic changes in microRNA expression during differentiation of rat cochlear progenitor cells in vitro. AB - OBJECTIVE: Cochlear progenitor cells could be used to explore the cochlea developmental mechanism and for cell replacement therapy in deafness. MicroRNAs are small, noncoding RNAs that could regulate the cell fate of stem cells, as well as cellular proliferation, differentiation and maturation. An expression profile analysis of microRNAs is necessary to understand their complex roles in differentiating cochlear progenitor cells. METHODS: The microRNAs microarray was used to analyze microRNA expression changes while differentiating cochlear progenitor cells. Quantitative real-time polymerase chain reaction was used to confirm and compare the results of the microarray and to detect the expression pattern of several microRNAs during the differentiation of neural stem cells. RESULTS: Nearly 100 microRNAs were identified from the microarray. Most showed changes in expression levels as cochlear progenitor cell differentiation progressed. The quantitative real-time polymerase chain reaction result demonstrated that the miR-183 family exhibits cell-specific expression in cochlear progenitor cells compared with neural stem cells. CONCLUSIONS: The temporal regulation of these microRNAs indicated that they might play different roles in differentiating cochlear progenitor cells, and that specific microRNAs might influence the cell fate determination of cochlear progenitor cells. PMID- 21621865 TI - Perception and negative effect of loneliness in a Chicago Chinese population of older adults. AB - This qualitative study aims to investigate the cultural understandings of loneliness, identify the contexts of loneliness, and to examine its effect on the health and well-being of U.S. Chinese older adults. Despite loneliness is one of the main indicators of well-being, little attention has been paid to understanding loneliness among immigrant older adults. This study utilizes both survey questionnaires and semi-structured focus group methods to investigate the feelings of loneliness among U.S. Chinese older adults. Based on interviews with 78 community-dwelling Chinese older adults in Chicago Chinatown, this community based participatory research study (CBPR) shows loneliness is common among U.S. Chinese older adults. It was frequently identified through emotional isolation and social isolation. Social, psychological and physical health factors could contribute to the experience of loneliness. In addition, the health of older adults with loneliness may be associated with worsening overall health, elder mistreatment, health behavior changes and increased healthcare utilizations. This study has implications for healthcare professionals, social services agencies and policy makers. Our findings point to the needs for healthcare professionals to be more alert of the association of loneliness and adverse health outcomes. Communities and social services agencies should collectively take a lead in reducing social isolation, improving intergenerational relationships, and increasing social networks and companionship for this group of vulnerable older adults. PMID- 21621864 TI - Zinc: new clues to diverse roles in brain ischemia. AB - Cerebral ischemia is a leading cause of morbidity and mortality, reflecting the extraordinary sensitivity of the brain to a brief loss of blood flow. A significant goal has been to identify pathways of neuronal injury that are selectively activated after stroke and may be amenable to drug therapy. An important advance was made nearly 25 years ago when Ca(2+) overload was implicated as a critical link between glutamate excitotoxicity and ischemic neurodegeneration. However, early hope for effective therapies faded as glutamate targeted trials repeatedly failed to demonstrate efficacy in humans. In a review in 2000 in this journal, we described new evidence linking a related cation, zinc (Zn(2+)), to neuronal injury, emphasizing sources and mechanisms of Zn(2+) toxicity. The current review highlights progress over the last decade, emphasizing mechanisms through which Zn(2+) ions (from multiple sources) participate together with Ca(2+) in different stages of cascades of ischemic injury. PMID- 21621866 TI - Long-term results of full-dose gemcitabine with radiation therapy compared to 5 fluorouracil with radiation therapy for locally advanced pancreas cancer. AB - PURPOSE: To retrospectively compare the efficacy and toxicity of full-dose gemcitabine based chemoradiotherapy (GemRT) versus 5-fluorouracil (5-FU) based chemoradiotherapy (5FURT) for locally advanced pancreas cancer (LAPC). METHODS: From January 1998 to December 2008, 93 patients with LAPC were treated either with 5FURT (n=38) or GemRT (n=55). 5FURT consisted of standard-field radiotherapy given concurrently with infusional 5-FU or capecitabine. GemRT consisted of involved-field radiotherapy given concurrently with full-dose gemcitabine (1000mg/m(2) weekly) with or without erlotinib. The follow-up time was calculated from the time of diagnosis to the date of death or last contact. RESULTS: Patient characteristics were not significantly different between treatment groups. The overall survival (OS) was significantly better for GemRT compared to 5FURT (median 12.5months versus 10.2months; 51% versus 34% at 1year; 12% versus 0% at 3years; 7% versus 0% at 5years, respectively; all P=0.04). The OS benefit of GemRT was maintained on subset analysis without concurrent erlotinib or with sequential gemcitabine (all P<0.05). The rates of distant metastasis, subsequent hospitalization, acute and late grade 3-5 gastrointestinal toxicities were not significantly different between the GemRT and 5FURT groups. CONCLUSIONS: GemRT was associated with an improved OS compared to standard 5FURT. This approach yielded long-term survivors and was not associated with increased hospitalization or severe gastrointestinal toxicity. PMID- 21621867 TI - Use of megavoltage cine-images for studying intra-thoracic motion during radiotherapy for locally advanced lung cancer. AB - BACKGROUND AND PURPOSE: Use of planning 4-dimensional CT (4DCT) scans often permits use of smaller target volumes for thoracic tumors but this assumes a reproducible pattern of motion during radiotherapy. We compared cranio-caudal (CC) motion on MV cine-images acquired during treatment with that seen on planning 4DCT. METHODS AND MATERIALS: A pre-programmable respiratory motion phantom and a software tool for motion assessment were used to validate the use of MV cine-images for motion detection. MV cine-images acquired in 20 patients with node-positive lung cancer were analyzed using the same software. Intra fraction CC motion on 6 MV cine-images from each patient was compared with CC motion on their planning 4DCT. RESULTS: Software-based motion measurement on MV cine-images from the phantom corresponded to actual motion. Mean CC motion of primary tumor, carina and hilus on 4DCT was 7.3mm (range 2-13.8mm), 6.8mm (1.8 21.2) and 11.0mm (4.2-15.1), respectively. Corresponding intra-fraction motion on MV cine was 4.1mm (0.6-13.6mm); 2.7mm (0-10mm) and 6.0mm (1.8-14.4mm), respectively. The tumor, hilus and carina could be tracked in 95%, 88% and 38% of the MV cine-images, respectively. CONCLUSIONS: Intra-fraction motion can be reliably measured using MV-cine images from a phantom. Motion discrepancies identified on MV cine-images can identify patients in whom planning 4DCT scans are not representative. PMID- 21621868 TI - Dose-volume analysis of locoregional recurrences in head and neck IMRT, as determined by deformable registration: a prospective multi-institutional trial. AB - BACKGROUND AND PURPOSE: Although IMRT for head and neck cancer is widely accepted, the implications of sparing normal tissue immediately adjacent to target volumes are not well known. MATERIALS AND METHODS: Between 2002 and 2007, 124 patients with head and neck cancer were treated with surgery and postoperative IMRT (n=79) or definitive RT (n=45). Locoregional recurrences were analyzed for location relative to target volumes, and dosimetry. RESULTS: With a median follow-up of 26.1months, a total of 16 locoregional recurrences were observed. The five-year actuarial locoregional disease-free survival was 82% [95% CI, 72-90%]. Analysis of 18 distinct sites of locoregional failure revealed that five of these failures were within the high dose clinical target volume (CTV), nine failures were at the margin of the CTV, and four recurrences were outside the CTV. The mean dose delivered to these recurrent volumes was 63.1 Gy [range: 57-68 Gy], while the mean dose to the coolest 1cc within each recurrence was 60.0 Gy [range: 51-67 Gy]. There were two periparotid recurrences observed. CONCLUSIONS: We observed excellent locoregional control rates overall. The majority of recurrences occur within high dose regions of the neck and not near the spared parotid glands. PMID- 21621869 TI - Trends in drug prescriptions to diabetic patients from 2000 to 2008 in Italy's Lombardy Region: a large population-based study. AB - OBJECTIVE: To analyze the prescribing patterns of antidiabetic and cardiovascular medications among diabetics in the most highly populated Italian Region, from 2000 to 2008. METHODS: Data were obtained from the Lombardy Region administrative health databases. The standardized prevalence of antidiabetic and cardiovascular drugs use was calculated within each study year. The prescription trends of initial treatment with antidiabetic drugs were also analyzed. RESULTS: From 2000 to 2008 there was an increase in the proportion of patients treated with biguanides (from 53.4% to 66.5%; p<0.0001) while those receiving sulfonylurea decreased (from 78.6% to 56.4%; p<0.0001). A sharp increase of metformin (as monotherapy) as initial treatment was also observed (from 15.2% to 48.8%; p<0.0001). The percentage of patients receiving renin-angiotensin system inhibitors, lipid-lowering drugs and antiplatelets increased between 2000 and 2008, from respectively 45.1% to 63.3%, 13.6% to 43.2% and 21.6% to 40.9 (p<0.0001). Multivariate analyses indicated that changes in prescriptions were statistically significant for both antidiabetic and cardiovascular drugs. CONCLUSION: This study documents progressive changes in the prescription of antidiabetic and cardiovascular drugs in accordance with guidelines. However, the use of metformin as first line therapy was still suboptimal and cardiovascular preventive strategies were only partially implemented in community practice. PMID- 21621870 TI - Automatic real-time detection of endoscopic procedures using temporal features. AB - Endoscopy is used for inspection of the inner surface of organs such as the colon. During endoscopic inspection of the colon or colonoscopy, a tiny video camera generates a video signal, which is displayed on a monitor for interpretation in real-time by physicians. In practice, these images are not typically captured, which may be attributed by lack of fully automated tools for capturing, analysis of important contents, and quick and easy retrieval of these contents. This paper presents the description and evaluation results of our novel software that uses new metrics based on image color and motion over time to automatically record all images of an individual endoscopic procedure into a single digitized video file. The software automatically discards out-patient video frames between different endoscopic procedures. We validated our software system on 2464 h of live video (over 265 million frames) from endoscopy units where colonoscopy and upper endoscopy were performed. Our previous classification method achieved a frame-based sensitivity of 100.00%, but only a specificity of 89.22%. Our new method achieved a frame-based sensitivity and specificity of 99.90% and 99.97%, a significant improvement. Our system is robust for day-to-day use in medical practice. PMID- 21621871 TI - Expression of p16 in non-small cell lung cancer and its prognostic significance: a meta-analysis of published literatures. AB - The prognostic value of p16 for survival of patients with non-small cell lung cancer (NSCLC) remains controversial. we performed a meta-analysis of the literatures in order to clarify its impact. Published studies in English were identified using an electronic search in order to aggregate the available survival results. To be eligible, a study had to have dealt with p16 protein assessment in NSCLC patients on the primary site and have reported survival data according to p16 expression. Twenty trials, comprising 1995 patients, provided sufficient information for the meta-analysis. Seventeen assessed any non-small cell lung cancer subtype, three assessed adenocarcinoma only. Eight identified high p16 expression as a favourable prognostic factor and one linked it with poor prognosis, Eleven trials were not significant. The overall combined hazard ratio (HR) calculated using a random-effects model suggested that high p16 expression has a favourable impact on survival in all NSCLC [0.69, 95% CI: 0.59-0.81]; The studies were categorized according to histology, disease stage and laboratory technique. The aggregated survival data showed a poor survival prognosis in squamous cell cancer with lower p16 expression [0.34, 95% CI: 0.13-0.91]. The adenocarcinoma subgroup had an HR of 0.91 [95% CI 0.76-1.10] without statistical significance. In early stage NSCLC (I-II), the aggregated HR was 0.42 [95% CI: 0.28-0.63], showing a worse survival for NSCLC with abnormal p16 expression; Results were significant with the HR of 0.61 [95% CI: 0.45-0.82] for five studies detecting p16 by immunohistochemistry with antibody clone G175-405. In conclusion, our meta-analysis shows that the p16 expression status is an independent prognostic factor in NSCLC, and this tendency is also found in the subgroups of squamous cell lung cancer and early stage NSCLC (I-II), but not in lung adenocarcinoma. PMID- 21621872 TI - Kaempferol from Semen cuscutae attenuates the immune function of dendritic cells. AB - Dendritic cells (DCs) are the critical leukocytes in regulating immune responses. Accordingly, DCs are the major target in the development of immunomodulators. In this study, we examined the effect of Semen cuscutae (SC), an important traditional Chinese medicine, on mouse bone marrow-derived DCs. We found that the n-butanol and methanol extracts of SC significantly suppressed LPS-stimulated DC activation. Several flavonoids were verified in the extracts using HPLC, and then kaempferol was identified as the major flavonoid in the methanol fraction of SC. Kaempferol was able to reduce cytokines and chemokines produced by LPS-stimulated DCs, and this reduction was not due to its cytotoxicity on DCs. In addition, DC maturation was impaired by kaempferol. Furthermore, kaempferol abrogated the ability of LPS-stimulated DCs to promote Ag-specific T cell activation, both in vitro and in vivo. Thus, we show for the first time that SC exhibits an immunosuppressive effect on DCs and that the active ingredient kaempferol attenuates DC function, which suggests that kaempferol has potential in the treatment of chronic inflammatory and autoimmune diseases. PMID- 21621873 TI - The study of the E-class SEPALLATA3-like MADS-box genes in wild-type and mutant flowers of cultivated saffron crocus (Crocus sativus L.) and its putative progenitors. AB - To further understand flowering and flower organ formation in the monocot crop saffron crocus (Crocus sativus L.), we cloned four MIKC(c) type II MADS-box cDNA sequences of the E-class SEPALLATA3 (SEP3) subfamily designated CsatSEP3a/b/c/c_as as well as the three respective genomic sequences. Sequence analysis showed that cDNA sequences of CsatSEP3 c and c_as are the products of alternative splicing of the CsatSEP3c gene. Bioinformatics analysis with putative orthologous sequences from various plant species suggested that all four cDNA sequences encode for SEP3-like proteins with characteristic motifs and amino acids, and highlighted intriguing sequence features. Phylogenetically, the isolated sequences were closest to the SEP3-like genes from monocots such as Asparagus virgatus, Oryza sativa, Zea mays, and the dicot Arabidopsis SEP3 gene. All four isolated C. sativus sequences were strongly expressed in flowers and in all flower organs: whorl1 tepals, whorl2 tepals, stamens and carpels, but not in leaves. Expression of CsatSEP3a/b/c/c_as cDNAs was compared in wild-type and mutant flowers. Expression of the isolatedCsatSEP3-like genes in whorl1 tepals together with E-class CsatAP1/FUL subfamily and B-class CsatAP3 and CsatPI subfamilies of genes, fits the ABCE "quartet model," an extended form of the original ABC model proposed to explain the homeotic transformation of whorl1 sepals into whorl1 tepals in Liliales and Asparagales plants such as C. sativus. This conclusion was also supported by the interaction of the CsatSEP3b protein with CsatAP1/FUL and CsatAP3 proteins. In contrast, expression of both B-class CsatAP3 and CsatPI genes and the C-class CsatAGAMOUS genes together with E-class CsatSEP3-like genes in carpels, without any phenotypic effects on carpels, raises questions about the role of these gene classes in carpel formation in this non grass monocot and requires further experimentation. Finally, taking advantage of the size and sequence differences in amplified genomic sequences of the triploid C. sativus and comparing them with the respective sequences from C. tomasii, C. hadriaticus and C. cartwrightianus, three putative wild-type diploid progenitor species, we examined the origin of CsatSEP3a sequence. PMID- 21621874 TI - [Screening for retinal detachment using wide-field retinal imaging]. AB - PURPOSE: The wide-field retinal imaging device (200 degrees ) (P200 Optomap Plus, Optos, Edinburgh, Scotland, UK), using a scanning laser ophthalmoscope (SLO), makes it possible to visualize a wide retinal area including the retinal periphery. The present study was designed to evaluate the ability to detect peripheral rhegmatogenous lesions using SLO wide-field image reading. METHODS: Between 2 November 2007 and 30 April 2008, all patients presenting with a retinal detachment were prospectively enrolled. Retinal ophthalmoscopy and drawing of retinal detachment, breaks, and holes were performed by a single retinal surgeon. SLO imaging was performed by an orthoptist and image reading by a senior resident. The orthoptist and the resident were both unaware of the results of the clinical examination. The extension of the retinal detachment in clock hours as well as the number of breaks and holes were compared between the retinal drawing and the image reading results. RESULTS: During the study period, 6053 images were taken, in 56 eyes with retinal detachment documented. Retinal breaks and holes were detected in seven of 10 cases with SLO image reading, only those situated superiorly between 11 and 1 o'clock and inferiorly between 5 and 7 o'clock were missed. Retinal detachments (one situated superiorly and one situated inferiorly) were detected in nine of 10 cases. CONCLUSIONS: The sensitivity of retinal detachment detection with the wide-field imaging device appears to be satisfactory for screening purposes, although it cannot replace retinal ophthalmoscopy. PMID- 21621875 TI - Length of stay and charges associated with health care-acquired bloodstream infections. AB - BACKGROUND: Although many studies have examined outcomes of health care associated bloodstream infections (HCABSIs), population-based estimates of length of stay (LOS) and costs have seldom been reported. OBJECTIVES: Our objective was to generate US national estimates of LOS and costs associated with HCABSIs using the 2003 National Inpatient Sample (NIS). METHODS: This study utilized a matched case-control design to estimate LOS and costs associated with HCABSIs based on the 2003 (NIS). A special set of ICD-9-CM codes was used to identify cases. A 1:1 matching procedure was used in which HCABSIs in patients were matched with uninfected patients based on age, sex, and admission diagnosis. We performed weighted analysis to construct population estimates and their standard deviations for LOS and total charges. RESULTS: After applying the case finding criteria, 113,436 HCABSI cases were identified. The weighted mean LOS for HCABSIs cases was 16.0 days compared with 5.4 days for the control group (P < .001). The weighted mean total charges for patients with HCABSIs were $85,813 ($110,183 US in 2010) compared with $22,821 ($29,302 US in 2010) for uninfected patients (P < .001). We estimated that, in 2003, HCABSIs potentially cost the US economy nearly $29 billion ($37.24 billion US in 2010). CONCLUSION: This study estimated the economic burden of HCABSIs on the US national economy. With some modifications, the annually published NIS data could be useful as a national surveillance tool for health care adverse events including HCABSIs. PMID- 21621876 TI - [Ibuprofen-induced cholestatic hepatitis]. PMID- 21621877 TI - [Cognitive impairment prevention: characteristics of memory workshops in Barcelona]. PMID- 21621878 TI - [Patients with mild cognitive impairment and a reduced CSF Abeta1-42 protein progress rapidly to Alzheimer's disease]. AB - INTRODUCTION: Some studies have shown that CSF amyloid-beta 1-42 (Abeta1-42), total tau (T-tau) and tau phosphorylated at threonine 181 (P-tau(181p)) proteins are useful diagnostic markers for distinguishing between clinically stable mild cognitive impairment (MCI) patients and those who will develop Alzheimers disease (AD). Our objective was to test the ability of this technique to discriminate in our cohort of MCI patients, according to the clinical outcome, one year after the lumbar puncture. MATERIAL AND METHODS: A total of 36 MCI patients were included from the local hospital memory clinic. Using INNO-BIA Alzbio-3 reagents from Innogenetics, we measured CSF Abeta1-42, T-tau and P-tau(181p) proteins, and calculated the T-tau/Abeta1-42 y P-tau(181p)/Abeta1-42 ratios. This project was approved by the local ethics committee. RESULTS: One year after the lumbar puncture, 14 MCI patients (38%) developed AD. These patients had lower Abeta1-42 protein levels (285.3 vs 377 ng/ml, P<.02) and higher P-tau(181p)/Abeta1-42 ratio (0,25 vs 0,16, p<.02) than the clinically stable patients. CONCLUSIONS: Our MCI patients with lower Abeta1-42 protein levels and an increased P-tau(181p) /Abeta1 42 ratio progressed quickly to AD. These results may help to identify those MCI patients with a poorer prognosis. PMID- 21621879 TI - Structural and functional neuroimaging in human prion diseases. AB - INTRODUCTION: Prion diseases are neurodegenerative disorders resulting from the accumulation of a misfolded isoform of the cellular prion protein (PrPc). They can occur as acquired, sporadic, or hereditary forms. Although prion diseases show a wide range of phenotypic variations, pathological features and clinical evolution, they are all characterised by a common unfavourable course and a fatal outcome. REVIEW SUMMARY: Some variants, such as kuru, have practically disappeared, while others, for example the variant Creutzfeldt-Jakob (vCJD) or those attributable to iatrogenic causes, are still in force and pose a challenge to current medicine. There are no definitive pre-mortem diagnostic tests, except for vCJD, where a tonsil biopsy detects 100% of the cases. For this reason, diagnostic criteria dependent on statistical probability have had to be created. These require complementary examinations, such as an electroencephalogram (EEG) or the detection of 14-3-3 protein in cerebrospinal fluid (CSF). Only the pulvinar sign in magnetic resonance imaging (MRI) has been included as a vCJD diagnostic criterion. The present review discusses neuroimaging findings for each type of prion disease in patients with a definitive histopathological diagnosis. CONCLUSIONS: The aim is to define the usefulness of these complementary examinations as a tool for the diagnosis of this family of neurodegenerative diseases. PMID- 21621880 TI - Design, synthesis and biological evaluation of novel 4-thiazolidinones containing indolin-2-one moiety as potential antitumor agent. AB - A series of novel 4-thiazolidinone and indolin-2-one hybrid derivatives 5a-5s and 10a-10s have been designed and synthesized and their cytotoxic activities were evaluated in vitro against three human cancer cell lines including HT-29 (human colon cancer), H460 (human lung cancer), MDA-MB-231 (human breast cancer) by MTT assay. Several potent target compounds (5m, 5p, 5s, 10a, 10c-10g, 10m, 10p) were further evaluated against one cancer cell line SMMC-7721 (human liver cancer) and one normal cell line WI-38 (human fetal lung fibroblasts). Most of the prepared compounds exhibited significant antitumor activities against different human cancer cell lines. Compound 10c (IC(50) = 0.025 MUM, 0.075 MUM, 0.77 MUM, 1.95 MUM) was 52, 36, 4.8 and 3.3 times more active than Sunitinib (IC(50) = 1.3 MUM, 2.7 MUM, 3.7 MUM, 6.47 MUM) against HT-29, H460, MDA-MB-231 and SMMC-7721 cancer cell line, respectively. PMID- 21621881 TI - Angiotensin-I-converting enzyme inhibitory peptides: Chemical feature based pharmacophore generation. AB - A validated 3D pharmacophore model was generated for a series of ACE inhibitory peptides, which consisted of five features (two hydrophobic functions, two hydrogen bond acceptors, and a negative ionizable function). The built model was able to correctly predict the activity of known ACE inhibitors. The model was then used as query to search 3D databases of peptides. Three novel peptides (I, II and III) were synthesized and biologically evaluated in vitro. It appears that the in vitro activity of peptides I, II and III was consistent with their molecular modeling results. Our results provided confidence for the utility of the pharmacophore model to retrieve novel ACE inhibitory peptides with desired biological activity by virtual screening. PMID- 21621882 TI - Studies on quinones. Part 47. Synthesis of novel phenylaminophenanthridinequinones as potential antitumor agents. AB - In our search for potential anticancer agents, a series of 8- and 9-phenylamino 3,4-tetrahydro-phenanthridine-1,7,10(2H)-triones with substituent variations at 6 , 8- and 9-positions were prepared using a highly efficient sequence involving: a) solar photoacylation reactions of benzoquinone with arylaldehydes, b) one-pot procedure for the synthesis of 3,4-dihydrophenanthridine-1,7,10(2H)-trione intermediates from acylhydroquinones and c) highly regiocontrolled acid-induced amination reaction of phenanthridinequinones with phenylamines. The members of this series were in vitro evaluated using the MTT colorimetric method against one normal cell line and three human cancer cell lines. The SAR analysis indicates that the location of nitrogen substituents on the quinone nucleus, the presence of methyl, phenyl, furyl and thienyl groups at the 6-position and the aromatization of the angular cycloaliphatic ring of the phenylamino-3,4 tetrahydrophenanthridine-1,7,10(2H)-trione pharmacophore play key roles in the antitumor activity. PMID- 21621883 TI - Structure-based optimization of click-based histone deacetylase inhibitors. AB - Previously, we reported a click-chemistry based approach to the synthesis of a novel class of histone deacetylase (HDAC) inhibitors [1]. The lead compound NSC746457 was found to be as potent as SAHA (Vorinostat). Further optimization of NSC746457 by using the HDAC2-TSA crystal structure is described herein. Docking of NSC746457 into HDAC2 binding domain suggested that the hydrophobic residue Phe210 flanking the cap-group binding-motif could be exploited for structural optimization. Substitution on the methylene group of cinnamic cap region led to identification of more potent HDAC inhibitors: isopropyl derivative 5 and tert butyl derivative 6, with an IC(50) value of 22 nM and 18 nM, respectively. PMID- 21621884 TI - Synthesis, structure and cytotoxicity of 3-C, N, S, Se substituted benzo[b]selenophene derivatives. AB - Synthesis, molecular structure and cytotoxic activity of a series of 3-C, N, S, Se substituted benzo[b]selenophene derivatives on human fibrosarcoma HT-1080, mouse hepatoma MG-22A, and mouse fibroblasts 3T3 cell lines are described. The correlation between compound LD(50) 3T3 fibroblast cell line and HT-1080 morphology was shown. PMID- 21621885 TI - Molecular modelling studies on Arylthioindoles as potent inhibitors of tubulin polymerization. AB - The crucial role played by microtubules in the life of eukaryotic cell makes tubulin an important route for the anticancer therapy. The Arylthioindoles (ATIs) along with the corresponding ketone and methylene compounds are potent tubulin assembly inhibitors. We are here reporting the result of a series of docking and molecular dynamics experiments on this series of compounds. The results obtained from our in silico studies not only provided us with an insight on the nature of the binding of the ATIs to tubulin, but were also at the core of the design of a new series of potent inhibitors of tubulin polymerization. PMID- 21621886 TI - Tolerability and efficacy of a low-volume enteral supplement containing key nutrients in the critically ill. AB - BACKGROUND & AIMS: To compare early supplementation with antioxidants and glutamine using a low-volume enteral supplement containing key nutrients to an energy adjusted standard elementary diet and to investigate its effect on clinical efficacy and tolerability in critically ill patients with sepsis/SIRS. The primary endpoints were length of stay in the ICU and sufficient enteral feed. METHODS: This was a randomized, prospective, single-blind, controlled study in 58 critically ill patients (56.9% male, mean age 46.7 years, mean APACHE II score 21.6). They received either a low-volume enteral supplement containing key nutrients or a diluted standard nutrition solution. After 10 or 14 days inflammatory parameters, catecholamine need, and maximal enteral delivery were determined. RESULTS: Patients receiving a low-volume enteral supplement containing key nutrients did not reach sufficient enteral feed more often than controls (76 vs. 62%, respectively, p = 0.17). The difference in vitamin E and selenium uptake was higher in the treatment group than controls (12.4 vs. 3.7 and 54.7 vs. 16.3, respectively, p <= 0.011). Parameters such as fever, antibiotic treatment, artificial ventilation, and death were comparable. This was also true for days of ICU or hospital stay (33 +/- 23 and 49 +/- 34 days, respectively). CONCLUSIONS: The low-volume enteral supplement containing key nutrients was well tolerated and led to a better vitamin E and selenium supply. However, it did not affect length of ICU or hospital stay. Further studies are necessary to determine which disease-specific subgroups may benefit from this supplementation or which group may be harmed. PMID- 21621887 TI - What you eat is what you are -- a role for polyunsaturated fatty acids in neuroinflammation induced depression? AB - As essential polyunsaturated fatty acids (PUFAs) influence both inflammatory and depressive disorders, nutrition related treatment methods deserve great research interest. However, currently biological mechanisms underlying the depression modulating effects of the PUFA Omega-3 (omega-3) and Omega-6 (omega-6) derived eicosanoids (central nervous system messengers) are not fully established. Depression related naturally occurring cell death (apoptosis) is thought to be mediated by excitotoxicity and free radicals that appear in the brain immediately following any inflammatory or ischemic damage, and increases the likelihood of clinically defined depression. This review explores the hypothesis that the interaction between omega-6 and omega-3 derived eicosanoids plays a central role in control over apoptosis linked with inflammation and inflammation-driven depression, via regulation of apoptosis inducing factors including excitotoxicity and free radicals. PMID- 21621888 TI - Floodplain methylmercury biomagnification factor higher than that of the contiguous river (South River, Virginia USA). AB - Mercury biomagnification on the South River floodplain (Virginia, USA) was modeled at two locations along a river reach previously modeled for methylmercury movement through the aquatic trophic web. This provided an opportunity to compare biomagnification in adjoining trophic webs. Like the aquatic modeling results, methylmercury-based models provided better prediction than those for total mercury. Total mercury Food Web Magnification Factors (FWMF, fold per trophic level) for the two locations were 4.9 and 9.5. Methylmercury FWMF for the floodplain locations were higher (9.3 and 25.1) than that of the adjacent river (4.6). Previous speculation was not resolved regarding whether the high mercury concentrations observed in floodplain birds was materially influenced by river prey consumption by riparian spiders and subsequent spider movement into the trophic web of the adjacent floodplains. Results were consistent with a gradual methylmercury concentration increase from contaminated floodplain soil, to arthropod prey, and finally, to avian predators. PMID- 21621889 TI - Geographic variation in cardioprotective antihypertensive medication usage in dialysis patients. AB - BACKGROUND: Despite their high risk of adverse cardiac outcomes, persons on long term dialysis therapy have had lower use of antihypertensive medications with cardioprotective properties, such as angiotensin-converting enzyme (ACE) inhibitors/angiotensin receptor blockers (ARBs), beta-blockers, and calcium channel blockers, than might be expected. We constructed a novel database that permits detailed exploration into the demographic, clinical, and geographic factors associated with the use of these agents in hypertensive long-term dialysis patients. STUDY DESIGN: National cross-sectional retrospective analysis linking Medicaid prescription drug claims with US Renal Data System core data. SETTING & PARTICIPANTS: 48,882 hypertensive long-term dialysis patients who were dually eligible for Medicaid and Medicare services in 2005. FACTORS: Demographics, comorbid conditions, functional status, and state of residence. OUTCOMES: Prevalence of cardioprotective antihypertensive agents in Medicaid pharmacy claims and state-specific observed to expected ORs of medication exposure. MEASUREMENTS: Factors associated with medication use were modeled using multilevel logistic regression models. RESULTS: In multivariable analyses, cardioprotective antihypertensive medication exposure was associated significantly with younger age, female sex, nonwhite race, intact functional status, and use of in-center hemodialysis. Diabetes was associated with a statistically significant 28% higher odds of ACE-inhibitor/ARB use, but congestive heart failure was associated with only a 9% increase in the odds of beta-blocker use and no increase in ACE-inhibitor/ARB use. There was substantial state-by-state variation in the use of all classes of agents, with a greater than 2.9-fold difference in adjusted-rate ORs between the highest and lowest prescribing states for ACE inhibitors/ARBs and a 3.6-fold difference for beta blockers. LIMITATIONS: Limited generalizability beyond study population. CONCLUSIONS: In publicly insured long-term dialysis patients with hypertension, there were marked differences in use rates by state, potentially due in part to differences in Medicaid benefits. However, geographic characteristics also were associated with exposure, suggesting clinical uncertainty about the utility of these medications. PMID- 21621890 TI - Severe metabolic alkalosis in a hemodialysis patient. AB - We present a patient with end-stage kidney disease receiving hemodialysis therapy who developed severe metabolic alkalosis secondary to vomiting. This case illustrates the important differences in pathogenesis, diagnosis, and management of this common acid-base disorder in patients without kidney function. The diagnostic approach and management strategy for metabolic alkalosis are discussed, highlighting the special issues to be considered in dialysis patients. PMID- 21621891 TI - Medial frontal event-related potentials and reward prediction: do responses matter? AB - Medial frontal event-related potentials (ERPs) following rewarding feedback index outcome evaluation. The majority of studies examining the feedback related medial frontal negativity (MFN) employ active tasks during which participants' responses impact their feedback, however, the MFN has been elicited during passive tasks. Many of the studies examining the MFN show enhanced effects when an error in reward prediction occurs (i.e. expected rewards are not delivered). To clarify the roles of reward prediction error and active responding in producing the MFN, the current study employed a reward prediction design with active and passive task blocks. Following the presentation of a reward predictor, participants (active task) or the computer (passive task) indicated whether participants would receive the outcome associated with a stimulus presented on the left or right of the reward predictor. The MFN was largest when the trial outcome was worse than predicted and this effect was enhanced when the participant, rather than the computer, made the choice. These results show that both reward prediction error and active choice impact the neural system of outcome monitoring with the largest MFN when the individual's decision led to the negative outcome. PMID- 21621892 TI - [An introduction to competing risks analysis]. AB - The need to develop treatments and/or programs specific to a disease requires the analysis of outcomes to be specific to that disease. Such endpoints as heart failure, death due to a specific disease, or control of local disease in cancer may become impossible to observe due to a prior occurrence of a different type of event (such as death from another cause). The event which hinders or changes the possibility of observing the event of interest is called a competing risk. The usual techniques for time-to-event analysis applied in the presence of competing risks give biased or uninterpretable results. The estimation of the probability of the event therefore needs to be calculated using specific techniques such as the cumulative incidence function introduced by Kalbfleisch and Prentice. The model introduced by Fine and Gray can be applied to test a covariate when competing risks are present. Using specific techniques for the analysis of competing risks will ensure that the results are unbiased and can be correctly interpreted. Full English text available from: www.revespcardiol.org. PMID- 21621893 TI - Arterial line in prehospital emergency settings - A feasibility study in four physician-staffed emergency medical systems. AB - BACKGROUND AND AIM OF THE STUDY: Arterial lines are widely used in operating rooms, critical care and emergency departments. Although invasive arterial blood pressure monitoring and arterial blood gas analysis are prehospitally available, the use of arterial lines in the field remains an exception. This study evaluates the feasibility, indications and therapeutic consequences of prehospital arterial line insertion. METHODS: Prospective observational study in four physician staffed emergency medical systems (EMS), documenting patient status, indications, location of puncture, number of tries and time for puncture and therapeutic consequences. RESULTS: During the one-year observation period, arterial line placement succeeded in 115 (83.9%) of 137 patients. The median time for successful arterial cannulation was 2 min (IQR 1, 3 min; range: 30-600s), for preparing the invasive blood pressure monitoring 3 min (IQR 2, 4 min, range: 30 600s). Main indications were cardiopulmonary resuscitation (36.5%), post resuscitation care (16.8%), respiratory insufficiency (24.1%) and unconsciousness (22.6%). Therapeutic consequences depended on whether the EMS was equipped with a blood gas analyzer or not and were, overall, reported in 51.3% of patients: fluids, vasoactive or antihypertensive therapy, correction of ventilation or acidosis. No complications occurred during the prehospital phase. CONCLUSION: The insertion of arterial lines is feasible under prehospital conditions, without delaying or complicating patient care. Indications originating from intrahospital use are also valid in the field. In particular when combined with arterial blood gas measurement, the use of arterial lines often leads to important therapeutic consequences. PMID- 21621894 TI - New pets, new allergies. PMID- 21621895 TI - The impact of deep disease on surgical treatment of endometriosis. AB - OBJECTIVE: To compare the difficulty of surgery in patients with and without deeply infiltrating endometriosis. STUDY DESIGN: Prospective cohort study performed in one hospital specialized in the surgical treatment of endometriosis. 193 consecutive patients undergoing excision of all visible endometriosis by laparoscopy (176 patients, 91.2%) or by laparotomy (17 patients, 8.2%). The duration of surgery, the number of operations, the number of day-surgery operations, the need to operate with a surgeon, the ability to perform complete excision during one operation, and the ability to perform operation by laparoscopy were compared in patients with and without deep lesions. RESULTS: The mean duration of surgery was 192 (SD 96), and 76 (SD 41)min in patients with and without deep lesions (p<0.001). Ureterolysis (66% vs. 20%, p<0.001), division of adhesions (92% vs. 69%, p<0.001), and hysterectomy (32% vs., 8%, p<0.001), were more often performed on patients with deep lesions. 41 patients (42%) with deep lesions, and 1 patient (1%) without deep lesions were operated with a surgeon (p<0.001). Day-surgery was less often performed on patients with deep lesions (11% vs. 45%, p<0.001). Complete excision during one operation was performed on 95% and on 97% of the patients with and without deep lesions (p=1.0). Complete excision was less often performed by laparoscopy in patients with deep lesions (79% vs. 95%, p<0.001). CONCLUSIONS: Surgical treatment of deep lesions is more demanding and time-consuming than surgical treatment of other types of endometriosis, and collaboration with a surgeon is often necessary. Complete excision during one operation is a realistic goal for endometriosis surgery, but it is significantly less often achievable by laparoscopy in patients with deep lesions than in patients without deep lesions. PMID- 21621896 TI - Refusal of emergency caesarean delivery in cases of non-reassuring fetal heart rate is an independent risk factor for perinatal mortality. AB - OBJECTIVE: To assess pregnancy outcome in women who initially refused medically indicated caesarean delivery (CD) in cases of non-reassuring fetal heart rate (FHR) patterns. STUDY DESIGN: A retrospective cohort study, comparing patients who refused and did not refuse caesarean delivery (CD) due to non-reassuring FHR tracings, was conducted. Deliveries occurred between the years 1988 and 2009 in a tertiary medical center. Multivariate analysis was performed to control for confounders. RESULTS: Out of 10,944 women who were advised to undergo CD due to non-reassuring FHR patterns, 203 women initially refused CD. Women refusing medical intervention tended to be older (30.6 +/- 6.9 vs. 28.29 +/- 6.1, P<0.001) and of higher parity (46.8% vs. 19.9% had more than 5 deliveries; P<0.001) as compared to the comparison group. Refusal of CD was significantly associated with adverse perinatal outcome. Using a multiple logistic regression model controlling for confounders such as maternal age, refusal of treatment was found as an independent risk factor for perinatal mortality (adjusted OR=3.3, C.I. 95% 1.8 5.9, P<0.001). A non-significant trend towards higher rates of adverse perinatal outcome was found when refusal latency time was longer than 20 min (OR=2, 95% CI 0.36-11.95; P=0.29). CONCLUSION: Refusal of CD in cases of non-reassuring FHR tracings is an independent risk factor for perinatal mortality. PMID- 21621897 TI - A systematic review and meta-analysis of randomized studies comparing misoprostol versus placebo for cervical ripening prior to hysteroscopy. AB - OBJECTIVE(S): Hysteroscopy is an effective method for examining the uterine cavity but has some limitations, including the occasional need for cervical dilatation. Misoprostol is routinely used for cervical dilatation in various procedures but has not gained wide acceptance for use before hysteroscopy. STUDY DESIGN: This review includes randomized controlled trials which compare the use of misoprostol versus placebo by different routes and doses before diagnostic or operative hysteroscopy. The MEDLINE database and the Cochrane Central Register of Controlled Trials were searched for articles published from January 1970 to April 2010. The outcome measures studied were related either to the facilitation of the hysteroscopic procedure (need for cervical dilatation, cervical width at the beginning of hysteroscopy, duration of the procedure and complications such as cervical tear and uterine perforation) or to the medication side-effects. With regard to side-effects, we studied the incidence of nausea, diarrhea, abdominal pain, bleeding, and fever. RESULTS: Vaginal misoprostol reduced the need for cervical dilatation in the total population of pre- and post-menopausal women to a statistically significant degree. In the subgroup of operative hysteroscopy the need for dilatation and the duration of the procedure were also significantly reduced. Most other outcomes relating to the facilitation of the procedure did not reach statistical significance. The side effects in the misoprostol group were significantly more frequent than in the placebo group. CONCLUSION(S): There is insufficient evidence to recommend the routine use of misoprostol before every hysteroscopy. As the lack of serious benefit from misoprostol is unlikely to be due to type II error, its use should be reserved for selected cases. PMID- 21621898 TI - sFlt-1 and PlGF levels in a patient with mirror syndrome related to cytomegalovirus infection. PMID- 21621899 TI - Ultrasonographic evaluation and anti-mullerian hormone levels after laparoscopic stripping of unilateral endometriomas. AB - OBJECTIVE: To evaluate the ovarian reserve after laparoscopic stripping of unilateral endometriomas by comparing the operated and non-operated ovaries. STUDY DESIGN: Bilateral ovarian volumes, antral follicle counts, and stromal blood flows were assessed by ultrasonography and anti-mullerian hormone (AMH) levels were analysed in 36 patients who had undergone laparoscopic cystectomy for unilateral ovarian endometrioma. RESULTS: Mean antral follicle counts (AFC) of the operated side ovaries were significantly lower on the second postoperative day (3.1 +/- 2.4 vs 5.2 +/- 3.7; p<0.05) and in the third month (3.7 +/- 2.1 vs 6.4 +/- 2.7; p<0.05). Pulsatility indices of the operated ovaries were significantly decreased on the second postoperative day (2.22 +/- 0.46 vs 1.76 +/ 0.51; p<0.05) while resistance indices were increased (0.81 +/- 0.06 vs 0.88 +/- 0.13; p<0.05). Doppler parameters had recovered and a non-significant decrease in AMH levels of the patients was recorded in the third month after surgery (2.03 +/ 0.41 ng/mL vs 1.95 +/- 0.62 ng/mL; p>0.05). CONCLUSION: Although laparoscopic stripping of endometriomas seems to affect the ovarian reserve in terms of AFCs, it does not have a significant negative impact at the end of three months as assessed by ovarian volumes, Doppler indices and AMH levels. Endometrioma surgery techniques are important in preserving normal functioning ovaries and further studies are necessary for optimising these surgical approaches. PMID- 21621900 TI - Spontaneous conception and live birth after gonadotoxic chemotherapy for an aggressive bilateral ovarian Burkitt's lymphoma. PMID- 21621901 TI - Periodontal disease and nitric oxide levels in low risk women with preterm labor. AB - OBJECTIVES: The aims of the study were to evaluate the possible association between periodontal disease (PD) and preterm labor (PTL), to assess whether the presence of periodontal disease in women with PTL increases the risk of preterm birth (PTB), and to evaluate the role of nitric oxide (NO) in this possible association. STUDY DESIGN: This study included 820 nulligravid women with low risk socioeconomic status: 400 cases with PTL and 420 controls with normal pregnancy, between 25 and 33 weeks+6 days of weeks' gestation. At enrollment, periodontal examination and detection of plasma, gingival and cervical NO levels were performed. We compared the periodontal status of the two groups; we detected the presence of PD and compared NO levels. We then followed the outcome of women with PTL and compared obstetric status and PTB rate between patients with and without PD. RESULTS: Logistic regression analysis revealed a strong association between PD and PTL (adjusted odds ratio: 2.83, 95% confidence interval (CI) 1.86 4.23; P<0.0001). Non parametric Mann-Whitney U-test demonstrated significant differences in gingival and cervical NO levels between women with PTL and controls (respectively, median 85.1 MUmol/L, interquartile range (i.r.) 51.2-177 vs median 50.5 MUmol/L, i.r. 34.5-109.65, P<0.0001; and median 102 MUmol/L, i.r. 53.05-182.7 vs median 38.9 MUmol/L, i.r. 32.87-46.1, P<0.0001). The ability of mean-gingival NO levels to predict PTL was examined by ROC curve analysis: the area under the curve was 0.817 (95% CI 0.774-0.854; P<0.0001). The cut-off level for the greatest sensitivity and specificity for mean-gingival NO levels was 116.04 MUmol/L (sensitivity 0.57, specificity 0.94). Positive and negative predictive values were, respectively, 90.65% and 69.73%. Among women with PTL, the risk of PTB increased in women with PD (adjusted relative risk: 3.85, 95% CI 2.11-6.06; P<0.0001). CONCLUSIONS: In this tested population, PTL is associated with PD. The presence of PD in women with PTL increases the risk of premature delivery. NO may be a new marker to explain this association. PMID- 21621902 TI - Promoter spacer DNA plays an active role in integrating the functional consequences of RNA polymerase contacts with -10 and -35 promoter elements. AB - Bacterial RNA polymerase (RNAP) interacts with conserved -10 and -35 promoter elements to recognize the promoter and to form an open complex in which DNA duplex around transcription start site melts. Using model DNA constructs (fork junction DNA) that mimic DNA structure found in the open complex we observed that the consequences of mutations in -10 promoter element for RNAP binding exhibited a striking dependence on the presence or absence of a functional -35 promoter element. A role of spacer DNA (a non-conserved DNA sequence connecting -10 and 35 promoter elements) in this phenomenon was probed with a series of fork junction DNA constructs containing perturbations to the spacer DNA. In the absence of a physical connection between the -10 and -35 DNA elements, or when 10 and -35 DNA elements were connected by a long flexible non-DNA linker, the dependence of RNAP interactions with -10 element on the strength of -35 element was lost. When these DNA elements were linked by a rigid DNA duplex or by a DNA duplex containing a short single-stranded gap, the coupling between the -10 and 35 binding activities was observed. These results indicated that promoter spacer DNA played an active role in integrating the functional consequences of RNA polymerase contacts with -10 and -35 promoter element. This role likely involves physical deformation of the spacer occurring in parallel with promoter melting as shown by Fluorescence Resonance Energy Transfer (FRET) experiments with the probes incorporated into spacer DNA. PMID- 21621903 TI - Equilibrium and kinetic studies of protein cooperativity using urea-induced folding/unfolding of a Ubq-UIM fusion protein. AB - Understanding the origins of cooperativity in proteins remains an important topic in protein folding. This study describes experimental folding/unfolding equilibrium and kinetic studies of the engineered protein Ubq-UIM, consisting of ubiquitin (Ubq) fused to the sequence of the ubiquitin interacting motif (UIM) via a short linker. Urea-induced folding/unfolding profiles of Ubq-UIM were monitored by far-UV circular dichroism and fluorescence spectroscopies and compared to those of the isolated Ubq domain. It was found that the equilibrium data for Ubq-UIM is inconsistent with a two-state model. Analysis of the kinetics of folding shows similarity in the folding transition state ensemble between Ubq and Ubq-UIM, suggesting that formation of Ubq domain is independent of UIM. The major contribution to the stabilization of Ubq-UIM, relative to Ubq, was found to be in the rates of unfolding. Moreover, it was found that the kinetic m-values for Ubq-UIM unfolding, monitored by different probes (far-UV circular dichroism and fluorescence spectroscopies), are different; thereby, further supporting deviations from a two-state behavior. A thermodynamic linkage model that involves four states was found to be applicable to the urea-induced unfolding of Ubq-UIM, which is in agreement with the previous temperature-induced unfolding study. The applicability of the model was further supported by site-directed variants of Ubq UIM that have altered stabilities of Ubq/UIM interface and/or stabilities of individual Ubq- and UIM-domains. All variants show increased cooperativity and one variant, E43N_Ubq-UIM, appears to behave very close to an equilibrium two state. PMID- 21621904 TI - Potential fresh water saving using greywater in toilet flushing in Syria. AB - Greywater reuse is becoming an increasingly important factor for potable water saving in many countries. Syria is one of the most water scarce countries in the Middle East. However, greywater reuse is still not common in the country. Regulations and standards for greywater reuse are not available. Recently, however, several stakeholders have started to plan for greywater reuse. The main objective of this paper is to evaluate the potential for potable water saving by using greywater for toilet flushing in a typical Syrian city. The Sweida city in the southern part of Syria was chosen for this purpose. Interviews were made in order to reflect the social acceptance, water consumption, and the percentage of different indoor water uses. An artificial wetland (AW) and a commercial bio filter (CBF) were proposed to treat the greywater, and an economic analysis was performed for the treatment system. Results show that using treated greywater for toilet flushing would save about 35% of the drinking water. The economic analyses of the two proposed systems showed that, in the current water tariff, the payback period for AW and CBF in block systems is 7 and 52 years, respectively. However, this period will reduce to 3 and 21 years, respectively, if full water costs are paid by beneficiaries. Hence, introducing artificial wetlands in order to make greywater use efficient appears to be a viable alternative to save potable water. PMID- 21621905 TI - Hydraulic management of a soil moisture controlled SDI wastewater dispersal system in an Alabama Black Belt soil. AB - Rural areas represent approximately 95% of the 14000 km(2) Alabama Black Belt, an area of widespread Vertisols dominated by clayey, smectitic, shrink-swell soils. These soils are unsuitable for conventional onsite wastewater treatment systems (OWTS) which are nevertheless widely used in this region. In order to provide an alternative wastewater dosing system, an experimental field moisture controlled subsurface drip irrigation (SDI) system was designed and installed as a field trial. The experimental system that integrates a seasonal cropping system was evaluated for two years on a 500-m(2) Houston clay site in west central Alabama from August 2006 to June 2008. The SDI system was designed to start hydraulic dosing only when field moisture was below field capacity. Hydraulic dosing rates fluctuated as expected with higher dosing rates during warm seasons with near zero or zero dosing rates during cold seasons. Lower hydraulic dosing in winter creates the need for at least a two-month waste storage structure which is an insurmountable challenge for rural homeowners. An estimated 30% of dosed water percolated below 45-cm depth during the first summer which included a 30-year historic drought. This massive volume of percolation was presumably the result of preferential flow stimulated by dry weather clay soil cracking. Although water percolation is necessary for OWTS, this massive water percolation loss indicated that this experimental system is not able to effective control soil moisture within its monitoring zone as designed. Overall findings of this study indicated that soil moisture controlled SDI wastewater dosing is not suitable as a standalone system in these Vertisols. However, the experimental soil moisture control system functioned as designed, demonstrating that soil moisture controlled SDI wastewater dosing may find application as a supplement to other wastewater disposal methods that can function during cold seasons. PMID- 21621906 TI - The EORTC tables overestimate the risk of recurrence and progression in patients with non-muscle-invasive bladder cancer treated with bacillus Calmette-Guerin: external validation of the EORTC risk tables. AB - BACKGROUND: European Organization for Research and Treatment of Cancer (EORTC) risk tables only included 171 patients treated with bacillus Calmette-Guerin (BCG) for non-muscle-invasive bladder cancer (NMIBC). OBJECTIVE: To evaluate the external validity of the EORTC tables in patients with NMIBC treated with BCG over 5-6 mo. DESIGN, SETTING, AND PARTICIPANTS: Data on 1062 patients treated with BCG were analyzed. MEASUREMENTS: Discrimination was assessed using the concordance index (c-index) and the prognostic separation index (PSEP). For calibration, probabilities of recurrence and progression obtained with the EORTC risk tables in our series were compared with those reported by the EORTC. RESULTS AND LIMITATIONS: With respect to the discriminative ability of the EORTC model, c index was similar to those reported in the EORTC series for recurrence. However, c-indices for progression in our series were lower than c-indices reported by Sylvester et al. [1]. Although PSEP in our series was lower than in the EORTC series for recurrence at 1 yr, similar results were found at 5 yr. Regarding progression, PSEP in our series was lower than in the EORTC series. Whilst a successful stratification of recurrence and progression probability at 1 and 5 yr was achieved using the EORTC tables in our series, model calibration showed lower risks of recurrence than those reported by Sylvester et al. [1] in all groups. For progression, lower risks were found in higher-risk groups. There are some limitations in the present study. A different distribution of patients was found, with higher proportions of primary grade 3 T1 tumors and tumors in situ than in the EORTC series. An additional limitation is that prior recurrence of the EORTC table was not included in our parameters. Consequently, two separate analyses were performed for recurrence. CONCLUSIONS: The EORTC model successfully stratified recurrence and progression risks in our cohort. However, the discriminative ability of the EORTC tables decreased in our patients for progression. Moreover, these tables overestimated risks of recurrence and progression after BCG therapy. PMID- 21621907 TI - Safety of presurgical targeted therapy in the setting of metastatic renal cell carcinoma. AB - BACKGROUND: In patients with metastatic renal cell carcinoma (mRCC), the timing of systemic targeted therapy in relation to cytoreductive nephrectomy (CN) is under investigation. OBJECTIVE: To evaluate postoperative complications after the use of presurgical targeted therapy prior to CN. DESIGN, SETTING, AND PARTICIPANTS: A retrospective review of all patients who underwent a CN at The University of Texas M.D. Anderson Cancer Center from 2004 to 2010 was performed. Inclusion in this study required documented evidence of mRCC, with treatment incorporating CN. INTERVENTIONS: Patients receiving presurgical systemic targeted therapy prior to CN were compared to those undergoing immediate CN. MEASUREMENTS: Complications were assessed using the modified Clavien system for a period of 12 mo postoperatively. RESULTS AND LIMITATIONS: Presurgical therapy was administered to 70 patients prior to CN (presurgical), while 103 patients had an immediate CN (immediate). A total of 232 complications occurred in 57% of patients (99 of 173). Use of presurgical systemic targeted therapy was predictive of having a complication>90 d postoperatively (p=0.002) and having multiple complications (p=0.013), and it was predictive of having a wound complication (p<0.001). Despite these specific complications, presurgical systemic targeted therapy was not associated with an increased overall complication risk on univariable or multivariate analysis (p=0.064 and p=0.237) and was not predictive for severe (Clavien >=3) complications (p=0.625). This study is limited by its retrospective nature. As is inherent to any retrospective study reporting on complications, we are limited by reporting bias and the potential for misclassification of specific complications. CONCLUSIONS: Despite an increased risk for specific wound-related complications, overall surgical complications and the risk of severe complications (Clavien >=3) are not greater after presurgical targeted therapy in comparison to upfront cytoreductive surgery. PMID- 21621908 TI - If a partial nephrectomy could be done safely for a renal tumor, would radical nephrectomy be considered malpractice? PMID- 21621909 TI - Familial renal cell carcinoma from the Swedish Family-Cancer Database. AB - BACKGROUND: Reliable data on familial risks are important for clinical counselling and cancer genetics. OBJECTIVE: To evaluate familial risks for renal cell carcinomas (RCC) through parental and sibling probands in the largest available dataset. DESIGN, SETTING, AND PARTICIPANTS: This study examined the Swedish Family-Cancer Database on 12.2 million individuals, which contains families with parents and offspring. Cancer data were retrieved from the Swedish Cancer Registry for the years 1961-2008, including 8513 patients with RCC. MEASUREMENTS: Familial risk for offspring was defined through standardised incidence ratios (SIRs) and adjusted for many variables, including a proxy for smoking and obesity. RESULTS AND LIMITATIONS: The familial risk for RCCs was 1.75 when a parent and 2.61 when a sibling was diagnosed with any kidney cancer. Also, RCCs were shown to be associated with prostate cancer (PCa) when parents or parents and siblings were diagnosed with PCa. Among siblings, the associations of RCC with melanoma, non-Hodgkin's lymphoma, and urinary bladder and papillary thyroid tumours were found. None of the results differed significantly after excluding the families with cancer pathognomonic of a von Hippel-Lindau (VHL) disease. Limitations of this study include the small number of familial cases (229 familial cases). CONCLUSIONS: The present analysis showed a high familiarity for RCC, and recessive effects may be important for familial aggregation of RCC. As a novel association, offspring RCC was in excess when parents or parents and siblings were diagnosed with PCa. There is familial clustering beyond VHL and the recent low-risk gene that probably explains a small proportion of the observed familial clustering. PMID- 21621910 TI - Objective measures of renal mass anatomic complexity predict rates of major complications following partial nephrectomy. AB - BACKGROUND: The association between tumor complexity and postoperative complications after partial nephrectomy (PN) has not been well characterized. OBJECTIVE: We evaluated whether increasing renal tumor complexity, quantitated by nephrometry score (NS), is associated with increased complication rates following PN using the Clavien-Dindo classification system (CCS). DESIGN, SETTING, AND PARTICIPANTS: We queried our prospectively maintained kidney cancer database for patients undergoing PN from 2007 to 2010 for whom NS was available. INTERVENTIONS: All patients underwent PN. MEASUREMENTS: Tumors were categorized into low- (NS: 4-6), moderate- (NS: 7-9), and high-complexity (NS: 10-12) lesions. Complication rates within 30 d were graded (CCS: I-5), stratified as minor (CCS: I or 2) or major (CCS: 3-5), and compared between groups. RESULTS AND LIMITATIONS: A total of 390 patients (mean age: 58.0 +/- 11.9 yr; 66.9% male) undergoing PN (44.6% open, 55.4% robotic) for low- (28%), moderate- (55.6%), and high-complexity (16.4%) tumors (mean tumor size: 3.74 +/- 2.4 cm; median: 3.2 cm) from 2007 to 2010 were identified. Tumor size, estimated blood loss, and ischemia time all significantly differed (p<0.0001) between groups; patient age, body mass index (BMI), and operative time were comparable. When stratified by CCS, minor and major complication rates for all patients were 26.7% and 11.5%, respectively. Minor complication rates were comparable (26.6 vs. 24.9 vs 32.8%; p=0.45), whereas major complication rates differed (6.4 vs. 11.1 vs. 21.9%; p=0.009) among tumor complexity groups. Controlling for age, gender, BMI, type of surgical approach, operative duration, and tumor complexity, prolonged operative time (odds ratio [OR]: 1.01; confidence interval [CI], 1.0-1.02) and high tumor complexity (OR: 5.4; CI, 1.2-24.2) were associated with the postoperative development of a major complication. Lack of external validation is a limitation of this study. CONCLUSIONS: Increasing tumor complexity is associated with the development of major complications after PN. This association should be validated externally and integrated into the decision-making process when counseling patients with complex renal tumors. PMID- 21621911 TI - Additional evidence for improved functional outcomes following robot-assisted radical prostatectomy. PMID- 21621912 TI - Relapses and remissions of hypokalaemic periodic paralysis and multiple sclerosis in the same patient. PMID- 21621913 TI - Clinical outcome of relapsing remitting multiple sclerosis among Hong Kong Chinese. AB - BACKGROUND: Clinical outcome of Chinese relapsing remitting multiple sclerosis (RRMS) patients is uncertain. AIM: To study the long-term clinical outcome of Chinese RRMS patients. METHOD: RRMS patients with duration of 10 years or longer followed up in our hospital is retrospectively studied. RESULTS: 61 RRMS patients (75% female) were studied. Their mean symptom onset age was 25.9 years and mean duration was 20.6 years (range 10-33); 36% patients had received beta-interferon and 30% azathioprine. Their mean EDSS scores were 3.3 (range 1-7) and 4.7 (range 1-8) at 10 years and latest follow-up (mean duration 20.6 years) respectively. At 10 years, 30% patients had EDSS score <=2, 34% EDSS 2.5-3.5, 20% EDSS 4.0-5.5 and 16% >=6; 18% developed SPMS. At latest follow-up, 15% patients had EDSS <=2, 20% EDSS 2.5-3.5, 19% EDSS 4.0-5.5 and 46% >=6.0; 53% developed SPMS. The median time from symptom onset to EDSS 6 was 22 years. No differences were detected in demographic characteristics, presenting neurological features, number of attacks in first 2 years, neuroradiological findings and disease modifying therapies between patients with EDSS <6 and >=6 at ten years. EDSS scores at 10 years and latest follow-up were similar for patients who had received beta-interferon and those who had not. CONCLUSION: Hong Kong Chinese RRMS patients may have worse long-term clinical outcome than Caucasian patients. PMID- 21621914 TI - Extraction of metals from spent hydrotreating catalysts: physico-mechanical pre treatments and leaching stage. AB - The present paper is focused on physico-mechanical pre-treatments of spent hydrotreating catalysts aimed at concentration of at least one of the valuable metals contained in such secondary raw material. In particular, dry Ni-Mo and Co Mo as well as wet Ni-Mo catalysts were used. Flotation, grain size separation and attrition processes were tested. After that, a rods vibrating mill and a ball mill were used to ground the catalysts in order to understand the best mechanical pre-treatment before leaching extraction. The results showed that flotation is not able to concentrate any metals due to the presence of coke or other depressant compounds. The particle size separation produces two fractions enriched in Mo and Co when dry Co-Mo catalyst is used, whereas attrition is not suitable as metals are uniformely distributed in rings' volume. Roasting at 550 degrees C and vibrating grinding are the most suitable pre-treatments able to produce fractions easily leached by NaOH and H(2)SO(4) after grain size separation. PMID- 21621915 TI - Destruction and formation of PCDD/Fs in a fluidised bed combustor co-incinerating automotive shredder residue with refuse derived fuel and wastewater treatment sludge. AB - During an eight day trial automotive shredder residue (ASR) was added to the usual waste feed of a Fluidized Bed Combustor (FBC) for waste-to-energy conversion; the input waste mix consisted of 25% ASR, 25% refuse-derived fuel (RDF) and 50% wastewater treatment (WWT) sludge. All inputs and outputs were sampled and the concentration of the 17 PCDD/Fs with TEF-values was determined in order to obtain "PCDD/F fingerprints". The ASR contained approximately 9000 ng PCDD/Fs/kg(DW), six times more than the RDF and 10 times more than the WWT sludge. The fingerprint of ASR and RDF was dominated by HpCDD and OCDD, which accounted for 90% of the total PDDD/F content, whereas the WWT sludge contained relatively more HpCDFs and OCDF (together 70%). The flue gas cleaning residue (FGCR) and fly and boiler ash contained approximately 30,000 and 2500 ng PCDD/Fs/kg(DW), respectively. The fingerprints of these outputs were also dominated by HpCDFs and OCDF. The bottom ash contained only OCDD and OCDF, in total 8 ng PCDD/Fs/kg (DW). From the comparison of the bottom ash fingerprints with the fingerprints of the other output fractions and of the inputs, it could be concluded that the PCDD/Fs in the waste were destroyed and new PCDD/Fs were formed in the post combustion process by de novo synthesis. During the ASR-co incineration, the PCDD/F congener concentrations in the fly and boiler ash, FGCR and flue gas were 1.25-10 times higher compared to the same output fractions generated during incineration of the usual waste mix (70% RDF and 30% WWT sludge). The concentration of the higher chlorinated PCDD/Fs increased most. As these congeners have the lowest TEF-factors, the total PCDD/F output, expressed in kg TEQ/year, of the FBC did not increase significantly when ASR was co incinerated. Due to the relatively high copper levels in the ASR, the copper concentrations in the FBCs outputs increased. As copper catalysis the de novo syntheses, this could explain the increase in PCDD/F concentrations in these outputs. PMID- 21621916 TI - Adsorptive behaviour of mercury on algal biomass: competition with divalent cations and organic compounds. AB - Biosorption processes constitute an effective technique for mercury elimination. Sorption properties of native and acid-treated Sargassum muticum have been studied. Effect of pH, initial mercury concentration and contact time studies provided fundamental information about the sorption process. This information was used as the reference values to analyse mercury sorption under competition conditions. Saline effect has shown little influence in sorption, when only electrostatic modifications took place upon salt addition. On the contrary, if mercury speciation dramatically changed owing to the addition of an electrolyte, such as in the case of chloride salt, very large modifications in mercury sorption were observed. Competition with other divalent cations or organic compounds has shown little or none effect on mercury, indicating that a different mechanism is taking place during the removal of these pollutants. Finally, continuous flow experiments have clearly shown that a reduction process is also taking place during mercury removal. This fact is not obvious to elucidate under batch sorption experiments. Scanning Electron Microscopy analysis of the surface of the materials show deposits of mercury(I) and metallic mercury which is indicative of the reduction process proposed. PMID- 21621917 TI - Enhanced tolerance and remediation of anthracene by transgenic tobacco plants expressing a fungal glutathione transferase gene. AB - Plants can be used for remediation of polyaromatic hydrocarbons, which are known to be a major concern for human health. Metabolism of xenobiotic compounds in plants occurs in three phases and glutathione transferases (GST) mediate phase II of xenobiotic transformation. Plants, although have GSTs, they are not very efficient for degradation of exogenous recalcitrant xenobiotics including polyaromatic hydrocarbons. Hence, heterologous expression of efficient GSTs in plants may improve their remediation and degradation potential of xenobiotics. In the present study, we investigated the potential of transgenic tobacco plants expressing a Trichoderma virens GST for tolerance, remediation and degradation of anthracene-a recalcitrant polyaromatic hydrocarbon. Transgenic plants with fungal GST showed enhanced tolerance to anthracene compared to control plants. Remediation of (14)C uniformly labeled anthracene from solutions and soil by transgenic tobacco plants was higher compared to wild-type plants. Transgenic plants (T(0) and T(1)) degraded anthracene to naphthalene derivatives, while no such degradation was observed in wild-type plants. The present work has shown that in planta expression of a fungal GST in tobacco imparted enhanced tolerance as well as higher remediation potential of anthracene compared to wild-type plants. PMID- 21621918 TI - Studies on the selective Am3+ transport, irradiation stability and surface morphology of polymer inclusion membranes containing Cyanex-301 as carrier extractant. AB - Transport behaviour of Am(3+) across cellulose triacetate (CTA) based polymer inclusion membranes (PIM) containing Cyanex-301 (bis(2,4,4 trimethylpentyl)dithiophosphinic acid) as the carrier extractant and tri-n-butyl phosphate (TBP) or 2-nitrophenyloctylether (NPOE) as the plasticizer was investigated from different feed and strip conditions. The TBP plasticized membrane resulted back transport of Am when alpha-hydroxy iso-butyric acid was used as the complexing agent in the strip phase while no such effect was seen when ethylene diamine tetraacetic acid (EDTA) was used as the complexant. Effect of varying Cyanex-301 concentration and bipyridyl (bipy) concentration on Am transport was also investigated. Long term reusability of the membrane was studied by measuring the permeability coefficient (P) after exposing the PIMs to a maximum gamma ray dose of ~ 200 kGy. The surface morphology of the membranes was analyzed by atomic force microscopy and the roughness parameter was correlated to transport efficiency. PMID- 21621919 TI - Definition of a short-cut methodology for assessing earthquake-related Na-Tech risk. AB - Na-Tech (Natural and Technological) refers to industrial accidents triggered by natural events such as storms, earthquakes, flooding, and lightning. Herein, a qualitative methodology for the initial assessment of earthquake Na-Tech risk has been developed as a screening tool to identify which situations require a much more expensive Quantitative Risk Analysis (QRA). The proposed methodology, through suitable Key Hazard Indicators (KHIs), identifies the Na-Tech risk level associated with a given situation (i.e., a process plant located in a given territory), using the Analytical Hierarchy Process as a multi-criteria decision tool for the evaluation of such KHIs. The developed methodology was validated by comparing its computational results with QRA results that involved Na-Tech events previously presented in literature. PMID- 21621920 TI - Analyses of genes encoding Theileria parva p104 and polymorphic immunodominant molecule (PIM) reveal evidence of the presence of cattle-type alleles in the South African T. parva population. AB - Restriction fragment length polymorphism analysis of PCR products (PCR-RFLP) and sequencing of the variable region of the p104 and PIM genes was performed on samples obtained from South African T. parva parasites originating from cattle on farms with suspected theileriosis and from buffalo. p104 and PIM PCR-RFLP profiles similar to those of the T. parva Muguga stock, an isolate that causes ECF in Kenya, were obtained from three of seven cattle samples collected on a farm near Ladysmith in KwaZulu-Natal Province. Amino acid sequences of the p104 and PIM genes from two of these samples were almost identical to the T. parva Muguga p104 and PIM sequences. This result supports findings from a recent p67 study in which p67 alleles similar to those of the T. parva Muguga stock were identified from the same samples. While these results suggest the presence of a cattle-derived T. parva parasite, reports of cattle-to-cattle transmission could not be substantiated and ECF was not diagnosed on this farm. Although extensive diversity of p104 and PIM gene sequences from South African T. parva isolates was demonstrated, no sequences identical to known cattle-type p104 and PIM alleles were identified from any of the buffalo T. parva samples analyzed. 'Mixed' PIM alleles containing both cattle- and buffalo-type amino acid motifs were identified for the first time, and there appeared to be selection of cattle-type and 'mixed'-type PIM sequences in the cattle samples examined. PMID- 21621921 TI - Dose titration of sericea lespedeza leaf meal on Haemonchus contortus infection in lambs and kids. AB - The objective of three experiments was to determine the impact of supplementing sericea lespedeza (Lespedeza cuneata; SL) in three concentrations in a loose or pelleted diet on gastrointestinal nematodes (GIN) in small ruminants. Experiments on lambs were conducted at the USDA, Agricultural Research Service in Booneville, AR (Exp. 1) and at Louisiana State University in Baton Rouge, LA (Exp. 2); an experiment on goat kids occurred at University of Maryland-Eastern Shore (Exp. 3). Exp. 1 used crossbred hair sheep lambs naturally infected with GIN that were randomly allocated to diets containing 0, 25, 50, and 75% SL diets (n=11 or 12/diet). Exp. 2 consisted of Haemonchus contortus-inoculated crossbred wool breed lambs that were blocked by gender and FEC and randomly assigned to 0, 25, 50, or 75% SL diet (n=8/diet). Fecal egg counts (FEC) and blood packed cell volume (PCV) were not influenced by SL supplementation in Exp. 1 and 2. Exp. 3 consisted of naturally GIN infected Boer crossbred goat kids in individual pens. Kids were blocked by FEC and randomly allotted to treatments of 0, 20, 40, or 60% SL with 9-13 goats/diet. The more SL fed, the greater the reduction in FEC (P<0.001). There was an increase in PCV in SL fed goats (P<0.001). Larval speciation at the end of the experiment indicated that feces from control animals produced 43% H. contortus larva while 20, 40 and 60% SL resulted in 39%, 35% and 31% H. contortus larvae, respectively. Feeding dried SL may be less effective in lambs than kids, though concurrent studies must be conducted to confirm this. PMID- 21621922 TI - The effect of nematode administration on canine atopic dermatitis. AB - Canine atopic dermatitis is a common disease and is considered as an animal model of the human disease. Immunomodulation by helminths is reported in several species. The aim of this study was to determine whether nematodes have an immunomodulatory effect on atopic dermatitis in dogs. In the pilot study, 12 atopic dogs were infected with either embryonated eggs of Trichuris vulpis (500 and 2500 eggs in 3 dogs each) or L3 larvae of Uncinaria stenocephala (100, 500 and 2500 eggs in 2 dogs each), respectively, for 3 months. Pruritus was evaluated with visual analogue scales and clinical lesions with the canine atopic dermatitis extent and severity index (CADESI). Skin biopsies were obtained for histopathology at the beginning and end of the study. In the subsequent placebo controlled, double-blinded, randomised study, 21 dogs received either 2500 embryonated T. vulpis eggs or placebo and were evaluated similarly. In addition, allergen-specific serum IgE concentrations were determined. All dogs in the pilot study improved in their lesion scores, most in their pruritus scores. The cutaneous inflammatory infiltrate did not change significantly. In the subsequent randomised study, there was no significant difference between placebo and Trichuris administration in regard to pruritus or CADESI. IgE concentrations also did not change significantly. Infection with T. vulpis did not significantly change clinical signs of canine atopic dermatitis. PMID- 21621923 TI - Mechanism of action of demethylating and immune modulatory agents--discussion. PMID- 21621924 TI - Development and validation of the Alcohol Identity Implicit Associations Test (AI IAT). AB - Alcohol identity is the extent to which an individual perceives drinking alcohol to be a defining characteristic of his or her self-identity. Although alcohol identity might play an important role in risky college drinking practices, there is currently no easily administered, implicit measure of this concept. Therefore we developed a computerized implicit measure of alcohol identity (the Alcohol Identity Implicit Associations Test; AI-IAT) and assessed its reliability and predictive validity in relation to risky college drinking practices. One hundred forty-one college students completed the AI-IAT. Again 3- and 6-months later, we administered the AI-IAT and indices of engagement in risky college drinking practices. A subset of participants also completed the previously-validated implicit measure of alcohol identity. Scores on the AI-IAT were stable over time, internally consistent, and positively correlated with the previously-validated measure of alcohol identity. Baseline AI-IAT scores predicted future engagement in risky college drinking practices, even after controlling for standard alcohol consumption measures. We conclude that the AI-IAT reliably measures alcohol identity, a concept that appears to play an important role in risky college drinking practices. PMID- 21621925 TI - The fusion of lipid droplets is involved in fat loss during cooking of duck "foie gras". AB - Fat loss during cooking of duck "foie gras" is the main quality issue in processing plants. To better understand this phenomenon, a histological and ultrastructural study was conducted. The aim was to characterize changes in lipid droplets of duck "foie gras" related to fat loss during cooking. Ten fatty livers were sampled before and after cooking and prepared for optical and transmission electron microscopy. In raw livers, the lipid droplets were nearly spherical while after cooking, they were larger and lost their spherical shape. We also observed a decrease in the number of droplets after cooking, probably due to droplet fusion caused by the heat treatment. Before cooking, there were fewer lipid droplets and a higher osmium tetroxyde staining intensity in the fatty liver, which later gave a lower technological yield. Fat loss during cooking was higher when there was more fusion of lipid droplets before cooking. PMID- 21621926 TI - Mitochondrial UCP4 and bcl-2 expression in imprints of breast carcinomas: relationship with DNA ploidy and classical prognostic factors. AB - Mitochondria are the bioenergetic and metabolic centers of cells and play an important role in the regulation of cell death. The mitochondrial apoptosis pathway is controlled by the bcl-2 protein family. Overexpression of mitochondrial uncoupling protein 4 (UCP4) can promote proliferation and inhibit apoptosis and differentiation. Imprint smears obtained from 124 tumors were studied immunocytochemically, and results were correlated with prognostic markers. There were 112 ductal and 12 lobular carcinomas. The positivity of UCP4 was correlated with lymph node metastases (p=0.005), positive ER and PR expression (p<0.0001 for both), as well as positivity for p53 (p<0.0001) and Ki 67 (p<0.0001). Decreased expression of bcl-2 correlated with increased expression of UCP4 (p=0.001). Regarding DNA ploidy, UCP4 positivity was correlated with aneuploid tumors (p=0.002). Negative expression of bcl-2 was correlated with poorly differentiated carcinomas (p<0.0001), as well as with positive expression of p53 (p<0.0001) and Ki-67 (p<0.0001). Logistic regression revealed that ploidy and p53 expression had an impact on UCP4. These findings encourage future investigations regarding the potential role of UCPs not only into mechanisms underlying breast cancer, but also as a novel candidate to the design and development of more effective therapeutic strategies. PMID- 21621927 TI - A reassessment of the role of the radial forearm flap in upper extremity reconstruction. AB - Our understanding of the hemodynamic consequences of radial artery harvest to the upper extremity has changed our considerations when approaching soft tissue defects of the hand. A critical assessment of the donor site morbidity of radial forearm (and radial artery) harvest is necessary to discern the role this flap should have in our current and future reconstructive armamentarium. A review of the available data and discussion of its implications is provided. PMID- 21621928 TI - [Clinical manifestations of adult celiac disease]. AB - Celiac disease is an enteropathy due to gluten intake in genetically predisposed persons (HLA DQ2/DQ8). Celiac disease occurs in adults and children at rates approaching 1% of population in Europe and USA. Celiac disease is extremely various and anaemia, oral aphthous stomatis, amenorrhea or articular symptoms may be the only revealing symptoms. Diagnosis releases on evidence of histological villous atrophy in proximal small bowel and presence of specific serum antibodies. Treatment relies on eviction of gluten. Gluten free diet allows prevention of malignant complications such as small bowel adenocarcinoma and lymphoma and osteopenia. The main cause of resistance to gluten free diet is its bad observance. On the contrary, serious complications of celiac disease, such as clonal refractory celiac sprue and intestinal T-cell lymphoma need to be screen. PMID- 21621929 TI - The effects of combined acamprosate and integrative behaviour therapy in the outpatient treatment of alcohol dependence: a randomized controlled trial. AB - AIMS: The aim of this randomized, controlled, multisite trial was to evaluate the efficacy of combined treatment with integrative behaviour therapy (IBT) and acamprosate on drinking behaviour in detoxified alcohol-dependent patients. METHODS: A total of 371 patients were randomized to one of the three treatment conditions: IBT plus acamprosate, IBT plus placebo, or supportive counselling ('treatment as usual', TAU) plus acamprosate. The main outcome was success rate, i.e., rate of abstinence plus improvement according to the criteria of Feuerlein and Kufner (1989), at the end of the six-month treatment phase and at the subsequent six-month follow-up. Drinking status was validated by blood parameters (CDT, GGT, and MCV). Data were analyzed by an intent-to-treat model and missing data were classified as relapse. RESULTS: The success rates at the end of treatment under both TAU plus acamprosate (37.7%) and IBT plus placebo (48%) almost reached the levels derived from the literature. However, adding acamprosate to IBT did not result in the expected increase in success rate (IBT plus acamprosate: 47.6%), and success rates did not differ significantly between groups. Similarly, there was no significant difference between treatment success rates at follow-up. CONCLUSION: The results suggest that the combination of acamprosate and IBT is not more effective than treatment with either IBT or acamprosate alone. However, the two acamprosate conditions differed in success rate by about 10%, which might constitute a clinically relevant though statistically non-significant effect. PMID- 21621930 TI - Alteration of virulence factors and rearrangement of pAsa5 plasmid caused by the growth of Aeromonas salmonicida in stressful conditions. AB - Aeromonas salmonicida, a fish pathogen, is the causative agent of furunculosis. It was already shown that growing this bacterium in stressful conditions such as temperature above 22 degrees C might lead to virulence attenuation. Unfortunately, many veterinary microbiology services and reference centers still routinely cultivate A. salmonicida at 25 degrees C. Here we tested the presence of virulence factors by growth on specific medium as well as the integrity of the pAsa5 plasmid, which bears an important virulence factor, the type III secretion system (TTSS), by PCR analysis in twenty strains, most of which were grown at 25 degrees C in their laboratory of origin. The analysis revealed that strains, which encountered the more stressful growth conditions displayed the most frequent absence of A-layer protein and secreted proteolytic activity. Moreover, many strains had lost parts of the pAsa5 plasmid in which the TTSS region was almost always affected. To confirm the effect of stressful growth conditions on the plasmid, three strains with an intact pAsa5 were cultured at 25 degrees C for two weeks. A low but significant fraction of the tested colonies displayed pAsa5 rearrangements. The rearrangement always affected the TTSS region and led to a loss of virulence in the Dictyostelium discoideum co-culture assay. These results demonstrate that the instability of pAsa5 did not lead to its complete loss as previously proposed but to a more complex rearrangement phenomenon and emphasizes the necessity to grow A. salmonicida in appropriate conditions to preserve the complete virulence of the bacterium. PMID- 21621931 TI - Comparison of virulence gene profiles of Escherichia coli isolates from sows with coliform mastitis and healthy sows. AB - Coliform mastitis (CM) is not only a serious economical and animal welfare touching problem in dairy cattle, but also in sows after farrowing. Due to this disease, the essential adequate supply with colostrum for the growth and the health of the piglets is not ensured. Besides other influencing factors, Escherichia (E.) coli is of great importance as a causative agent of this multifactorial disease. In this study, E. coli isolates from milk samples of healthy and CM-affected sows were examined for the presence of virulence genes associated with extraintestinal E. coli strains, enterotoxigenic E. coli and other pathogenic E. coli. The isolated E. coli harbored mainly virulence genes of extraintestinal E. coli strains (especially fimC, ompA, traT, hra, kpsMTII, iroN). The virulence gene spectrum for both samples from CM-affected and healthy sows did not differ significantly. Particular virulence gene profiles of E. coli isolates from diseased sows were not detected. This study provides novel insights into the role of E. coli in association with mastitis in sows since it is the first time E. coli isolates from CM-affected sows' milk were analysed for virulence genes. Because there were no differences in the prevalence of E. coli and their virulence-associated genes between healthy and diseased sows, other causative factors seem to have greater influence on the pathogenesis of porcine CM. PMID- 21621932 TI - Functional analysis of luxS in Streptococcus suis reveals a key role in biofilm formation and virulence. AB - Streptococcus suis (SS) is an important pathogen of pigs, responsible for diverse diseases in swine and human. LuxS has been reported to play critical roles in both regulating various behaviors and interspecies quorum sensing in a large spectrum of bacteria. In this study, the luxS deletion mutant of SS was constructed using homologous recombination and its biofilm formation, hemolytic activity, cell adherence, virulence and expression of virulence factors were evaluated. Compared to the parental strain, the biofilm formation and hemolytic activity were significantly decreased in the luxS mutant. The addition of synthetic autoinducer 2 could complement the deficiencies of biofilm production in the mutant strain. Furthermore, its adherence to the HEp-2 cell line was dramatically decreased by 51% compared to the parental strain. Expressions of the known virulence genes gdh, cps, mrp, gapdh, sly, fbps and ef in the mutant strain were decreased by 0.66, 0.61, 0.45, 0.48, 0.29, 0.57 and 0.38, respectively, as quantified by real-time PCR. In a zebrafish infection model, the 50% lethal dose of the mutant strain was increased up to 10-fold. The findings demonstrated that the luxS gene deletion resulted in a significant decrease of bacterial biofilm formation, cell adhesion, hemolytic activity and transcription levels of many virulence genes in SS, and these factors may be associated with the attenuation of virulence in zebrafish. These results suggest that luxS may be involved in the interruption of bacterial communication and biofilm formation that contribute to the virulence of the bacterium. PMID- 21621933 TI - Uniform treatment thresholds for hyperbilirubinemia in preterm infants: background and synopsis of a national guideline. AB - BACKGROUND: To prevent severe hyperbilirubinemia and bilirubin neurotoxicity, the American Academy of Pediatrics' management guideline for hyperbilirubinemia in near term infants is used worldwide. A leading guideline for jaundiced preterm infants is lacking whereas the risk on severe hyperbilirubinemia is high in these infants. Our aim was to define uniform treatment thresholds for jaundiced preterm infants. In this article we present the history and a synopsis of this novel national guideline. STUDY DESIGN: A survey on guidelines for hyperbilirubinemia in preterm infants was sent to all Dutch Neonatal Intensive Care Units (NICUs). After comparison with international guidelines, a new consensus-based guideline was developed. RESULTS: Treatment thresholds of all 10 NICUs were based on Total Serum Bilirubin (TSB) and related to birth weight (n = 9) and gestational age (n = 1). NICUs used age-specific (n = 6) or fixed (n = 4) TSB-thresholds resulting in a large range of thresholds (maximal 170 MUmol/L for phototherapy and 125 MUmol/L for exchange transfusion). Acidosis, asphyxia, sepsis, active hemolysis and intraventricular hemorrhage were the most frequently used risk factors. Consensus was agreed upon TSB-based treatment thresholds, categorized in 5 birth weight groups and divided in high and low risk infants. CONCLUSION: There was no standardized care for jaundiced preterm infants in the Netherlands. In addition to the internationally used guideline for (near) term infants, a novel "consensus based" guideline for preterm infants with a gestational age of less than 35 weeks has been developed and implemented in the Netherlands. This guideline is approved and recommended by the Dutch Society of Pediatrics. PMID- 21621934 TI - Vaccine therapy with sipuleucel-T (Provenge) for prostate cancer. AB - As the most common malignancy among North American males, prostate cancer causes more than 30,000 deaths each year. After local and hormonal treatments, a great number of patients ultimately progressed to castrate-resistant prostate cancer (CRPC), in which chemotherapy provides a small survival advantage, but with significant toxicities. In the past decade, prostate cancer has become a target for several immunotherapeutic approaches. Sipuleucel-T (Provenge(r), or APC8015) is a novel cancer vaccine developed from autologous dendritic cells (DC) loaded with engineered fusion protein of prostatic acid phosphatase (PAP) and granulocyte-macrophage colony-stimulating factor (GM-CSF). Phase I and Phase II trials show that the vaccine is safe and effective in creating immune responses toward the fusion-protein target antigen, PAP-GM-CSF also call PA2024. Recent Phase III studies also demonstrated sipuleucel-T's efficacy in prolonging median survival in patients with CRPC, despite little or no effect on clinical disease progression or surrogates such as serum PSA kinetics. Subsequently, the United States Food and Drug Administration approved sipuleucel-T for the treatment of asymptomatic or minimally symptomatic CRPC in April 2010. Filings are projected with international regulatory agencies in 2011. While the development of sipuleucel-T provides an option for patients with early CRPC, it also introduces physicians and researchers to new unanswered questions regarding its optimal clinical use and questions about mechanism of action and combination and sequencing with other agents. PMID- 21621935 TI - Assessing the use of digital radiography and a real-time interactive pulmonary nodule analysis system for large population lung cancer screening. AB - RATIONALE AND OBJECTIVES: To assess the use of chest digital radiograph (DR) assisted with a real-time interactive pulmonary nodule analysis system in large population lung cancer screening. MATERIALS AND METHODS: 346 DR/CR patient studies with corresponding CT images were selected from 12,500 patients screened for lung cancer from year 2007 to 2009. Two expert chest radiologists established CT-confirmed Gold Standard of nodules on DR/CR images with consensus. These cases were read by eight other chest radiologists (participating radiologists) first without using a real-time interactive pulmonary nodule analysis system and then re-read using the system. Performances of participating radiologists and the computer system were analyzed. RESULTS: The computer system achieved similar performance on DR and CR images, with a detection rate of 76% and an average FPs of 2.0 per image. Before and after using the computer-aided detection system, the nodule detection sensitivities of the participating radiologists were 62.3% and 77.3% respectively, and the A(z) values increased from 0.794 to 0.831. Statistical analysis demonstrated statically significant improvement for the participating radiologists after using the computer analysis system with a P value 0.05. CONCLUSION: The computer system could help radiologists identify more lesions, especially small ones that are more likely to be overlooked on chest DR/CR images, and could help reduce inter-observer diagnostic variations, while its FPs were easy to recognize and dismiss. It is suggested that DR/CR assisted by the real-time interactive pulmonary nodule analysis system may be an effective means to screen large populations for lung cancer. PMID- 21621936 TI - Diversity of nodule-endophytic agrobacteria-like strains associated with different grain legumes in Tunisia. AB - This study represents the first report describing the genetic diversity of nodule endophytic agrobacteria isolated from diverse legumes and their phylogenetic relationships with the valid species of agrobacteria, as well as the non recognized genomospecies of the former Agrobacterium tumefaciens (Rhizobium radiobacter). The genetic diversity of a collection of 18 non-nodulating agrobacteria-like strains, previously isolated from root nodules of Vicia faba, Cicer arietinum and Phaseolus vulgaris from different geographical regions of Tunisia, was studied by REP-PCR and PCR-RFLP of the 16S-23S rDNA IGS, as well as by sequence analysis of the 16S rDNA and the housekeeping genes recA and atpD. The aim of the work was to study the genetic diversity of the different isolates and to check for any host-specificity. The results from the different techniques were congruent and suggested a specific interaction for P. vulgaris, whereas no specific endophytic interaction was observed for V. faba and C. arietinum. The phylogenetic analysis clearly indicated that some isolates were affiliated to R. radiobacter or to its non-recognized genomic species (genomovars G2, G4 and G9). However, the other isolates probably constitute new species within Rhizobium (Agrobacterium) and Shinella. PMID- 21621937 TI - The celC gene, a new phylogenetic marker useful for taxonomic studies in Rhizobium. AB - The celC gene codifies for a cellulase that fulfils a very significant role in the infection process of clover by Rhizobium leguminosarum. This gene is located in the celABC operon present in the chromosome of strains representing R. leguminosarum, Rhizobium etli and Rhizobium radiobacter whose genomes have been completely sequenced. Nevertheless, the existence of this gene in other species of the genus Rhizobium had not been investigated to date. In this study, the celC gene was analysed for the first time in several species of this genus isolated from legume nodules and plant tumours, in order to compare the celC phylogeny to those of other chromosomal and plasmidic genes. The results obtained showed that phylogenies of celC and chromosomal genes, such as rrs, recA and atpD, were completely congruent, whereas no relation was found with symbiotic or virulence genes. Therefore, the suitability and usefulness of the celC gene to differentiate species of the genus Rhizobium, especially those with closely related rrs genes, was highlighted. Consequently, the taxonomic status of several strains of the genus Rhizobium with completely sequenced genomes is also discussed. PMID- 21621938 TI - Isolation and characterization of Thermanaerothrix daxensis gen. nov., sp. nov., a thermophilic anaerobic bacterium pertaining to the phylum "Chloroflexi", isolated from a deep hot aquifer in the Aquitaine Basin. AB - A new strictly anaerobic thermophilic multicellular filamentous bacterium (0.2 0.3MUm*>100MUm), designated GNS-1(T), was isolated from a deep hot aquifer in France. It was non-motile, and stained Gram-negative. Optimal growth was observed at 65 degrees C, pH 7.0, and 2gL(-1) of NaCl. Strain GNS-1(T) was chemoorganotrophic fermenting ribose, glucose, galactose, arabinose, fructose, mannose, maltose, sucrose, xylose, raffinose, pyruvate, and xylan. Yeast extract was required for growth. The end products of glucose fermentation were lactate, acetate, CO(2), and H(2). The G+C content of the DNA was 57.6mol%. Its closest phylogenetic relative was Bellilinea caldifistulae with 92.5% similarity. Based on phylogenetic, genotypic and phenotypic characteristics, strain GNS-1(T) (DSM 23592(T), JCM 16980(T)) is proposed to be assigned to a novel species of a novel genus within the class Anaerolineae (subphylum I), phylum "Chloroflexi", Thermanaerothrix daxensis gen. nov., sp. nov. The GenBank accession number is HM596746. PMID- 21621939 TI - A one step genotypic identification of Lactococcus lactis subspecies at the species/strain levels. AB - A Lactococcus lactis subspecies-specific primer was designed based on their repetitive genome sequences. This primer enabled L. lactis subspecies to be identified simultaneously at both the species level and also the strain level. Based on studies using 70 strains of L. lactis and 60 strains of other non-target bacteria, the identification completely matched that obtained by the sequence of the 16S rRNA gene. However, inconsistency between phenotypic and genotypic characteristics was observed in some strains isolated from milk. PMID- 21621940 TI - Restoration of hepatocytes function following decompression therapy in extrahepatic biliary obstructed patients: metabolite profiling of bile by NMR. AB - Prolonged biliary obstruction and infection cause retention of biliary constituents in liver followed by hepatocyte dysfunction. Decompression therapy is important for both management and prognostic reasons and restores hepatocyte function. Quantitative metabolite profiling of bile using NMR spectroscopy at the time of obstruction and serially following decompression therapy using percutaneous transhepatic biliary drainage (PTBD) from nineteen patients with extrahepatic malignant biliary obstruction are presented. Based on detailed history, clinical condition, total leucocyte counts (TLC) and microbiological cultures of bile, patients were classified in two groups depending upon absence or presence of cholangitis. Statistical analysis was performed for comparison within each group using Wilcoxan sign square rank test. TLC and liver function tests indicated a trend towards recovery following decompression by one week. While on day 0 biliary constituents were undetectable in both the groups of patients, they increased significantly following one week of drainage with better recovery in patients with cholangitis compared to without. Free amino acids' signals were detected in all specimens starting from day 1 after decompression. This indicates disruption of blood-bile barrier during cholestasis and slow restoration of tight junction of hepatocytes following decompression leading to the appearance of biliary constituents in bile. Decompression therapy tends to restore biliary constituents with a prompt recovery in patients with cholangitis further supports such therapy for clinical management and outcome. To our knowledge this is the first report on the detection of amino acids in bile taken from common bile duct though they have been reported in hepatic bile. PMID- 21621941 TI - Quantitative liquid chromatographic determination of intact cisplatin in blood with microwave-assisted post-column derivatization and UV detection. AB - The anticancer agent cisplatin (cis-diamminedichloroplatinum(II), cis [PtCl2(NH3)2]) easily undergoes ligand-exchange reactions, resulting in mainly inactive Pt complexes. This paper presents a method for selective analysis of intact cisplatin in blood using LC and UV detection. Blood samples (hematocrit: 0.22-0.52) were spiked with cisplatin (final concentrations: 2.48 * 10-7 M-9.90 * 10-6 M) and subjected to centripetal ultrafiltration. The blood ultrafiltrate was separated (loop volume: 5 MUl) with a porous graphitic carbon column and a mobile phase of HEPES-buffer (pH 9.3). Prior to UV detection (344 nm), the eluate was mixed with sodium N,N-diethyldithiocarbamate (DDTC) in a microwave field (115 degrees C) in order to improve the UV absorptivity. Cisplatin eluted as a Pt-DDTC complex after 11.8 min. The peak area was influenced primarily by the hematocrit, the DDTC concentration, and the temperature and residence time in the microwave cavity. The method was robust and sensitive provided preparing a fresh DDTC solution each day and, at the end of a day's run, destroying DDTC remaining in the system. It offers the main advantages of high selectivity, sensitivity, and robustness, minimal sample processing, and the possibility to use small sample volumes. PMID- 21621942 TI - Exploring the community pharmacist's role in palliative care: focusing on the person not just the prescription. AB - OBJECTIVE: Changes in health care provision have led to an emphasis on providing end of life care within the home. community pharmacists are well positioned to provide services to community-based palliative care patients and carers. METHODS: A multiple qualitative case study design was adopted. A total of 16 focus groups and 19 interviews with pharmacists, nurses, general practitioners and carers were undertaken across metropolitan and regional settings in Western Australia, New South Wales, Queensland and Victoria. Data were analysed thematically using a framework that allowed similarities and differences across stakeholder groups and locations to be examined and compared. RESULTS: Three main themes emerged: effective communication; challenges to effective communication; and: towards best practice, which comprised two themes: community pharmacists' skills and community pharmacists' needs. DISCUSSION: A key component of the provision of palliative care was having effective communication skills. Although community pharmacists saw an opportunity to provide interpersonal support, they suggested that they would need to develop more effective communication skills to fulfil this role. CONCLUSION: There is clear need for continuing professional development in this area - particularly in communicating effectively and managing strong emotions. PRACTICE IMPLICATIONS: Community pharmacists are willing to support palliative care patients and carers but need education, support and resources. PMID- 21621943 TI - The contribution of the Medicines Use Review (MUR) consultation to counseling practice in community pharmacies. AB - OBJECTIVE: To understand the contribution of the Medicines Use Review consultation to counseling practice in community pharmacies. METHODS: Qualitative study involving ten weeks of observations in two community pharmacies and interviews with patients and pharmacy staff. RESULTS: 'Traditional' counseling on prescription medicines involved the unilateral transfer of information from pharmacist to patient. Over-the-counter discussions were initiated by patients and offered more scope for patient participation. The recently introduced MUR service offers new opportunities for pharmacists' role development in counseling patients about their medicines use. However, the study findings revealed that MUR consultations were brief encounters dominated by closed questions, enabling quick and easy completion of the MUR form. Interactions resembled counseling when handing out prescription medicines. Patients rarely asked questions and indeterminate issues were often circumvented by the pharmacist when they did. MURs did little to increase patients' knowledge and rarely affected medicine use, although some felt reassured about their medicines. Pragmatic constraints of workload and pharmacy organisation undermined pharmacists' capacity to implement the MUR service effectively. CONCLUSION: Pharmacists failed to fully realise the opportunity offered by MURs being constrained by situational pressures. PRACTICE IMPLICATIONS: Pharmacist consultation skills need to be reviewed if MURs are to realise their intended aims. PMID- 21621944 TI - Development and validation of the Medication-Related Consultation Framework (MRCF). AB - OBJECTIVES: To develop and test a framework for evaluating the consultation skills of practitioners undertaking medication-related consultations. METHODS: Key components of medication-related consultations were identified through a literature review and compiled to form an initial consultation skills framework. This was iteratively refined through consultation with experts (n=21) to form the Medication-Related Consultation Framework (MRCF). Psychometric testing was undertaken by analyzing pharmacists' (n=10) assessment of fifteen pre-recorded simulated consultations. RESULTS: The MRCF consisted of 46 consultation behaviors, grouped into five sections. Performance was rated at individual behavior, framework section and global consultation levels. The MRCF discriminated between good, satisfactory and poor consultations at the global rating level (p<0.01) with good test-retest reliability (rho=0.59-0.95) and moderate inter-assessor reliability (Kendall's W=0.67). There was also good internal consistency for the five sections (Cronbach's alphas=0.58-0.97). CONCLUSIONS: The MRCF demonstrated good psychometric properties at the global and section rating levels. Some inconsistencies in assessors' ratings of individual consultation behaviors were indentified, which may represent a future training need. PRACTICE IMPLICATIONS: The MRCF provides healthcare professionals with a patient-centered consultation structure, serving to identify medication-related needs and potentially support adherence. It also allows the quality of a practitioner's consultation to be evaluated. PMID- 21621945 TI - Australian community pharmacists' practice in complementary medicines: a structural equation modeling approach. AB - OBJECTIVE: This survey tested a model of factors influencing pharmacists' practice in relation to complementary medicines (CMs). METHODS: Self-administered questionnaires were used to collect data from 212 community pharmacists in New South Wales. Structural equation modeling with AMOS version 6.0, was used to model two practices in relation to CMs: an evaluation of appropriateness of CM use and the decision to sell. RESULTS: Pharmacists' perceptions of their responsibilities in ensuring the safe use of CMs predicted the comprehensiveness of an evaluation for appropriateness of CM use (P=0.002). When level of comprehensiveness of evaluation increased, pharmacists were less likely to sell CMs in situations where the use of the products was not considered appropriate (P=0.021). Pharmacists' confidence in their CM knowledge, attitudes towards CMs, concern about pharmacy income and pharmacists' characteristics did not significantly affect their practice. CONCLUSION: Pharmacists' acceptance of their counseling responsibility with respect to CMs was the strongest predictor of both comprehensiveness of an evaluation and the decision to sell a CM. PRACTICE IMPLICATIONS: Pharmacists must be encouraged to recognise and accept their responsibility in ensuring the safe and effective use of CMs. PMID- 21621946 TI - ABOP, the automatic patient information leaflet optimizer: evaluation of a tool in development. AB - OBJECTIVE: We developed a semi-automated leaflet optimizer (ABOP) to improve the readability of Dutch patient information leaflets (PILs). Our aim was to assess whether or not revisions made with ABOP have an effect on traceability and comprehension of PIL information. METHODS: Two leaflets (one for oxazepam and one for tetracycline) were revised with the ABOP tool and both the original versions and ABOP versions were tested for readability, according to the EC guideline. RESULTS: The ABOP version for oxazepam (p<.0001) scored significantly better than its original counterpart in readability tests. Although the results showed that the ABOP version of the tetracycline antibiotic outperformed the original version the difference in scores was not statistically significant. Terminology improvements had the greatest effect on information retrieval and comprehension. Improvements to instructions and warnings did not affect retrieval and comprehension scores. Overall improvements did not ensure full compliance with EC requirements, but results are not far off. CONCLUSION: This study shows that even though ABOP does not solve all text quality issues, it does largely prepare a PIL for readability testing. PRACTICE IMPLICATIONS: Using ABOP as a revision tool for PILs can optimize PIL quality and hence reduce readability testing time and costs. PMID- 21621947 TI - Asthma disease management-Australian pharmacists' interventions improve patients' asthma knowledge and this is sustained. AB - OBJECTIVE: To assess any improvements in knowledge of asthma patients after a tailored education program delivered by pharmacists and measure the sustainability of any improvements. To ascertain patients' perceptions about any changes in their knowledge. METHODS: Ninety-six specially trained pharmacists recruited patients based on their risk of poor asthma control. A tailored intervention was delivered to patients based on individual needs and goals, and was conducted at three or four time points over six months. Asthma knowledge was assessed at the beginning and end of the service, and six and 12 months after it had ended. Patients' perceptions of the impact of the service on their knowledge were explored qualitatively in interviews. RESULTS: The 96 pharmacists recruited 570 patients, 398 (70%) finished. Asthma knowledge significantly improved as a result of the service (7.65 +/- 2.36, n=561, to 8.78 +/- 2.14, n=393). This improvement was retained for at least 12 months after the service. Patients reported how the knowledge and skills gained had led to a change in the way they managed their asthma. CONCLUSION: Improvements in knowledge are achievable and sustainable if pharmacists used targeted educational interventions. PRACTICE IMPLICATIONS: Pharmacist educational interventions are an efficient way to improve asthma knowledge in the community. PMID- 21621948 TI - Communication with patients who are dispensed a first prescription of chronic medication in the community pharmacy. AB - OBJECTIVE: To analyze information and communication in the community pharmacy when patients collect a first dispensing for chronic medication. METHODS: In 3 pharmacies in the South-West of the Netherlands, counter-based communication with patients receiving a first dispensing for chronic medication was observed and audiotaped. Each contact was analyzed by two observers using an observation checklist. An 11-item questionnaire was given to the patients. RESULTS: 72 Patients were included. Only pharmacy technicians provided information. Average contact lasted approximately 2min. A computer checklist to support information giving was closely followed. Interactions were structured with introduction and closure. Mostly, information was given without exploring needs of the patient. Questioning showed open-ended questions (26%); check-questions and questions asking feedback (57%). A mean general score given was 5.8 (range 1-9). Of 46 responders who filled in a questionnaire, the majority felt that information was useful, clear and understandable. CONCLUSION: Our results show a concise and efficient way of information giving, closely related to a computer checklist. Technicians dominate the interaction. They ask few questions. Patients are satisfied about provided information and the contact. PRACTICE IMPLICATIONS: We suggest a more patient-centered way of communication to increase patients' participation and to meet patients' needs for information. PMID- 21621949 TI - The effect of numeracy on the comprehension of information about medicines in users of a patient information website. AB - OBJECTIVE: To investigate the relationship between numeracy and the accuracy of side effect risk estimation following the presentation of information about a medicine via the Cancer Research UK (CR-UK) patient information website. METHODS: 591 website users were presented with information in different formats about the risks of side effects from taking tamoxifen. Participants estimated the risk of each side effect, provided other subjective ratings about the information and completed a numeracy task. RESULTS: Regardless of presentation format, numeracy was correlated with the accuracy of three side effect risk estimates. People with cancer and tamoxifen users showed stronger correlations for all side effect estimates. In addition, numeracy was positively related to the perceived influence of the information on the decision to take the medicine and was negatively related to ratings of satisfaction with the information. CONCLUSION: People with a lower numeracy level make larger errors in interpreting medicines side effect risk information. PRACTICE IMPLICATIONS: Pharmacists, other health professionals and patient information websites should ensure they provide clear explanations of risk, particularly to people with low numeracy, and assess their understanding of those explanations. Future research into risk communication should take account of numeracy level, to investigate the impact of different formats. PMID- 21621951 TI - Beer promotes high levels of alcohol intake in adolescent and adult alcohol preferring rats. AB - Previous studies suggest that high levels of alcohol consumption can be obtained in laboratory rats by using beer as a test solution. The present study extended these observations to examine the intake of beer and equivalent dilute ethanol solutions with an inbred line of alcohol-preferring P rats. In Experiment 1, male adolescent P rats and age-matched Wistar rats had access to either beer or equivalent ethanol solutions for 1h daily in a custom-built lickometer apparatus. In subsequent experiments, adolescent (Experiment 2) and adult (Experiment 3) male P rats were given continuous 24-h home cage access to beer or dilute ethanol solutions, with concomitant access to lab chow and water. In each experiment, the alcohol content of the beer and dilute ethanol solutions was gradually increased from 0.4, 1.4, 2.4, 3.4, 4.4, 5 to 10% EtOH (vol/vol). All three experiments showed a major augmentation of alcohol intake when rats were given beer compared with equivalent ethanol solutions. In Experiment 1, the overall intake of beer was higher in P rats compared with Wistar rats, but no strain difference was found during the 1-h sessions with plain ethanol consumption. Experiment 1 also showed that an alcohol deprivation effect was more readily obtained in rats with a history of consuming beer rather than plain ethanol solutions. In Experiments 2 and 3, voluntary beer intake in P rats represented ethanol intake of 10-15 g/kg/day, among the highest reported in any study with rats. This excessive consumption was most apparent in adolescent rats. Beer consumption markedly exceeded plain ethanol intake in these experiments except at the highest alcohol concentration (10%) tested. The advantage of using beer rather than dilute ethanol solutions in both selected and nonselected rat strains is therefore confirmed. Our findings encourage the use of beer with alcohol-preferring rats in future research that seeks to obtain high levels of alcohol self-administration. PMID- 21621950 TI - Doctor, what's wrong with me? Factors that delay the diagnosis of colorectal cancer. AB - OBJECTIVE: To examine the influence of patient and physician communication factors on diagnostic delay (DD). METHODS: 242 patients diagnosed with colorectal cancer (CRC) in the past 6 months who experienced symptoms prior to diagnosis were administered a 2-h semi-structured qualitative interview to assess communication with health care provider and ease of access to care, among other factors. Patient-provided information was verified via review of medical records. RESULTS: The factors associated with DD>2 months included lower income (OR=0.56, p=0.03), having regular physician prior to receiving a cancer diagnosis (OR=2.52, p=0.03), having a physician who used temporizing communication strategies during the consultation (OR=2.41, p=0.02), receiving an initial alternate diagnosis (OR=3.36, p=0.02), experiencing referral delay (OR=3.61, p=<0.001), and experiencing follow-up delay of any kind (OR=3.32, p=0.01). CONCLUSION: Excellent communication skills that appropriately probe for relevant social and economic patient information, assist patients in distinguishing and elaborating on symptoms, and provide clear rationale and instructions for future steps, will speed along the diagnosis process and could be the difference between early and late stage CRC. PRACTICE IMPLICATIONS: Increased understanding of physician communication and practice styles that contribute to DD could have a positive impact on decreasing the morbidity and mortality from this disease. PMID- 21621952 TI - Impact of surgical and clinical factors on the pharmacology of intraperitoneal doxorubicin in 145 patients with peritoneal carcinomatosis. AB - BACKGROUND: Cytoreductive surgery (CRS) and hyperthermic intraperitoneal chemotherapy (HIPEC) are a combined treatment modality considered for selected patients with peritoneal carcinomatosis from colorectal and appendiceal cancer. Doxorubicin is a drug consistently used by our group in this clinical setting. The surgical and clinical factors that modify the pharmacokinetics of HIPEC may be important for the design of future perioperative chemotherapy regimens. MATERIALS AND METHODS: The patients included were 145 who had colorectal or appendiceal carcinomatosis resected using CRS prior to treatment with HIPEC with doxorubicin as part of a multidrug regimen. The effect of clinical and surgical factors on drug distribution after a single intraperitoneal bolus administration with doxorubicin was determined. RESULTS: The pharmacokinetics of 145 patients treated with intraperitoneal doxorubicin showed a 78 times greater exposure to peritoneal surfaces as compared to plasma. At 90 min 12% of the drug remained in the chemotherapy solution and 88% was retained in the body. The extent of visceral resection and peritonectomy increased the clearance of doxorubicin from the peritoneal space. A major resection of visceral peritoneal surface, a contracted peritoneal space, and an incomplete cytoreduction reduced drug clearance. CONCLUSIONS: Surgical and clinical factors may require modifications of chemotherapy administration. A large visceral resection and a contracted peritoneal space caused a reduced doxorubicin clearance. Total diffusion surface is an important determinant of doxorubicin pharmacokinetics. PMID- 21621953 TI - [One year implemention of the safe surgery checklist in France, what has been achieved so far, what could be improved?]. AB - Despite important advances accomplished during the last ten years, patient safety in the OR remains a subject high on everyone's priority list: healthcare professionals, organisations and of course, patients. In this setting, the French National Authority for Health (Haute Autorite de sante, HAS) conducted a study with the scientific societies of professionals working in the OR. This study resulted in the adaptation of a tool which has already demonstrated, in a convincing manner, its efficacy in reducing perioperative morbimortality: the World Health Organisation's (WHO) "Surgical Safety Surgery" checklist. In order to promote its, HAS integrated this important tool for improving patient safety into the framework of its certification process of health care organisations beginning in January 1, 2010. Additionally, the organisations receive HAS certification are partners for the programme's promotion. One year after its institution, HAS is undertaking its first evaluation of the lessons and perspectives from the checklist's utilisation. PMID- 21621954 TI - Cementless Metasul metal-on-metal total hip arthroplasties at 13 years. AB - Second-generation metal-on-metal bearings have been used since the late 1980s as alternative bearings to eliminate aseptic loosening due to polyethylene wear. The aim of the present study was to evaluate the long-term results of a series of Metasul (Zimmer GmbH, Winterthur, Switzerland) metal-on-metal total hip arthroplasty (THA). One hundred forty-nine cementless THAs with a 28-mm Metasul articulation were performed in 111 consecutive patients. The results were retrospectively reviewed at 13 years postoperatively. Clinical and radiographic evaluations and implant survivorship were performed. Seven hips (4.7%) were revised. The overall survivorship with revision for any reason as the end point was 0.94. The average Harris hip score was 91.4. Expansive osteolysis was found adjacent to the stem in 4 hips. Metal wear-related aseptic loosening was not the major reason for failure in our Metasul metal-on-metal THAs. Recurrent dislocation was the main reason for revision in our series. PMID- 21621955 TI - Irrigation and debridement for periprosthetic infections: does the organism matter? AB - Irrigation and debridement (I&D) is an attractive treatment alternative for periprosthetic joint infection (PJI). Irrigation and debridement failure rates average 64% (range, 10.5%-84%) and may be associated with causative organism type and virulence. The study objective was to compare revision rates for PJI caused by streptococcal organisms to other organisms treated with I&D. A multicenter retrospective cohort study of 200 consecutive PJIs treated with I&D was performed. Failure was defined as reoperation for PJI. Failure rate for streptococcal infections was 65% (20/31) compared with 71% (84/119) for other organisms. Failure rate for sensitive Staphylococcus was 72% (48/67) compared with a 76% (22/29) failure rate for resistant Staphylococcus. These results indicate that eradication rates of I&D for a streptococcal PJI are comparable with other causative organisms. Irrigation and debridement should play a limited role in the PJI treatment algorithm regardless of organism type. PMID- 21621956 TI - Computer-assisted femoral head-neck osteochondroplasty using a surgical milling device an in vitro accuracy study. AB - Surgical navigation might increase the safety of osteochondroplasty procedures in patients with femoroacetabular impingement. Feasibility and accuracy of navigation of a surgical reaming device were assessed. Three-dimensional models of 18 identical sawbone femora and 5 cadaver hips were created. Custom software was used to plan and perform repeated computer-assisted osteochondroplasty procedures using a navigated burr. Postoperative 3-dimensional models were created and compared with the preoperative models. A Bland-Altmann analysis assessing alpha angle and offset ratio accuracy showed even distribution along the zero line with narrow confidence intervals. No differences in alpha angle and offset ratio accuracy (P = 0.486 and P = 0.2) were detected between both observers. Planning and conduction of navigated osteochondroplasty using a surgical reaming device is feasible and accurate. PMID- 21621957 TI - Impaction bone grafting with proximal and distal femoral arthroplasty. AB - Capacious diaphyses and poor bone stock in revision arthroplasty can lead to fracture and poor component fixation. Impaction bone grafting can be performed in salvage reconstructions in cases with extensive circumferential bone loss. We present a consecutive series of patients who underwent proximal or distal femoral reconstructions in combination with impaction bone grafting. The average age was 62 years, and the mean follow-up was 36 months (range, 24-84). No revisions were performed for mechanical failure, and radiographs revealed no evidence of implant loosening. No patient complained of end-of-stem thigh pain. There were 3 failures: 1 for periprosthetic fracture, 1 for instability, and 1 for infection. Impaction bone grafting can be used in combination with proximal and distal femoral arthroplasty to successfully manage cases with extensive bone loss after failed hip and knee arthroplasty. The procedure reconstitutes necessary bone stock, improves stress shielding, and provides immediate fixation in capacious canals. PMID- 21621958 TI - A prospective randomized clinical trial comparing tibial baseplate fixation with or without screws in total knee arthroplasty: a radiographic evaluation. AB - In a randomized prospective study, we compared the use of tibial screws with screwless tibial baseplate fixation in uncemented total knee arthroplasty and report the results of 154 prostheses at a mean follow-up of 5 years. Patients were mobilized within 24 hours and continued with partial weight-bearing for up to 6 weeks postoperatively. There were no significant differences in radiologic results and revision rates. Radiographic outcomes do not seem to be influenced by the use of screws in tibial baseplate fixation of this prosthesis. The prosthesis studied has a large tibial keel and 4 adjunctive pegs that seem to provide adequate stability and allow bony ingrowth to take place. Longer follow-up will determine whether in this implant design, the use of tibial screws for cementless tibial baseplate fixation can be justified. PMID- 21621959 TI - A prospective comparison of warfarin to aspirin for thromboprophylaxis in total hip and total knee arthroplasty. AB - Current orthopedic practice requires consideration of contradictory recommendations regarding pulmonary embolism (PE) prevention among patients undergoing total hip arthroplasty (THA) and knee joint arthroplasty (TKA). A total of 696 consecutive patients underwent elective THA or TKA. Two hundred eighty-one patients received PE risk stratification per American Academy of Orthopaedic Surgeons guidelines. Of these patients, 152 standard-risk patients received aspirin, and 129 elevated-risk patients received warfarin. The comparator group of 415 patients received American College of Chest Physicians recommended warfarin without PE risk stratification. Primary study outcomes were symptomatic PE, deep venous thrombosis, major bleeding, and death. The rate of symptomatic PE and venous thromboembolism among standard-risk group patients receiving aspirin was greater than the comparator group (4.6% vs 0.7% and 7.9% vs 1.2%, respectively). Most events (16/18) occurred among patients undergoing TKA. Patients with total joint arthroplasty at standard risk for PE receiving aspirin had a higher rate of symptomatic PE and venous thromboembolism than did patients receiving anticoagulation. PMID- 21621960 TI - Midterm results of primary total hip arthroplasty using highly cross-linked polyethylene: minimum 7-year follow-up study. AB - The purpose of this study was to investigate the results of 113 total hip arthroplasties in 109 patients using highly cross-linked polyethylene (HXLPE) after midterm (minimum 7 years) follow-up retrospectively. The mean age at the time of operation was 57 years. Preoperative diagnosis was osteonecrosis in 81 hips and other diagnosis in 32 hips. There was no component loosening. Acetabular osteolysis was found in 12 hips (10.6 %). Mean linear HXLPE wear rate was 0.031 +/- 0.012 mm/y. We analyzed the relationship between HXLPE wear rate and several variables influencing HXLPE wear. Only the cup position was related with wear rate of HXLPE significantly (P < .05). The results of total hip arthroplasties using HXLPE showed excellent results with decreased wear rate and low incidence of osteolysis after midterm follow-up. PMID- 21621961 TI - Safety and efficacy of a rotating-platform, high-flexion knee design three- to five-year follow-up. AB - Our hypothesis was that a high-flexion rotating-platform posterior stabilized (RP PS) design could maximize range of motion (ROM) and improve deep-flexion activities. Eighty-seven consecutive patients (109 knees) with high-flexion RP-PS design were prospectively followed up for a minimum of 3 years. Radiographic and clinical outcomes were analyzed using Knee Society Score criteria and Western Ontario and McMaster Universities Osteoarthritis Index. Good to excellent clinical scores were achieved in 96% of the knees. There were no cases of infection, malalignment, loosening, osteolysis, or spinout. The mean preoperative ROM improved from 110.7 degrees to 124 degrees postoperatively. High-flexion activities such as squatting and kneeling were achieved in 62% and 60% of patients, respectively. High-flexion knee implant is safe and improves ROM and deep-flexion activities. Persistent pain is an important cause of dissatisfaction after total knee arthroplasty. PMID- 21621962 TI - Accuracy and precision of two computer-assisted methods of radiographic wear measurement in total hip arthroplasty. AB - Roentgen Monographic Analysis Tool (ROMAN) and Hip Analysis Suite (HAS) were used to analyze radiographs of a phantom hip model. Displacements of known magnitude and direction were produced using dial micrometers. Differences between the known displacement and the programs' reported displacement were compared. Hip Analysis Suite was superior with a median error of 0.075 mm (range, 0.019-0.205 mm) compared with 0.137 mm (range, 0.008-0.389 mm) for ROMAN (P = .002). Hip Analysis Suite was also more precise when evaluating intraobserver variability, with a standard deviation between radiographs of 0.007 mm (range, 0.002-0.009 mm), whereas ROMAN's standard deviation was 0.117 mm (range, 0.007-0.153 mm). Repeatability for HAS was 0.019 mm and 0.325 mm for ROMAN. Hip Analysis Suite was more accurate and precise than ROMAN under experimental conditions with digital radiographs. PMID- 21621963 TI - How do cancer patients receiving palliative care at home die? A descriptive study. AB - CONTEXT: Data regarding the circumstances of the process of death of terminally ill patients followed at home are lacking. OBJECTIVES: The aim of this study was to describe the characteristics and assess the circumstances of the process of death of terminally ill patients followed at home. METHODS: This was a prospective survey to assess the dying process of advanced cancer patients followed at home. Within a week after death, the principal caregiver was interviewed. Information from the palliative home care team and the caregiver about expectation of death, time of death, professional and nonprofessional people present at time of death, emergency admission to hospital, and administration of drugs to resuscitate was gathered. The principal clinical issues in the last two hours also were recorded. RESULTS: In total, 181 of 222 caregivers provided information. Most deaths were expected. Palliative home care team physicians and nurses visited the patient on the day of death but were occasionally present at the moment of death. More than three people were generally present at time of death. More than two-thirds of patients died peacefully, without apparent suffering, and 35.7% of them received palliative sedation before dying. In the last two hours, the most frequent clinical issues were ranked as death rattle, dyspnea, and agitation. In 10 cases, emergency drugs for resuscitation were administered. CONCLUSION: This study has shown how advanced cancer patients die at home and that palliative home care may be helpful in allowing a death at home, particularly when relatives are actively involved. PMID- 21621964 TI - Reliability and validity of a Thai version of the edmonton symptom assessment scale (ESAS-Thai). AB - CONTEXT: The Edmonton Symptom Assessment Scale (ESAS), created by the Edmonton Group in 1991, is an instrument assessing symptom control that is commonly used in palliative care. It asks patients to rate nine items on 11-point numeric rating scales. OBJECTIVES: The aim of this study was to translate the ESAS to Thai and validate its final version with transcultural adaptation for Thai palliative care patients. The original ESAS was translated into Thai following the process of cross-cultural adaptation of self-report measures' standard guidelines, including forward translation, synthesis of the translation, back translation, cross-cultural adaptation, and pretesting. METHODS: This cross sectional study was first undertaken with 44 patients with advanced cancer in an inpatient setting, which led to the final version. The reliability and validity of the final version was then examined in a sample of 37 cancer patients in the outpatient department at Ramathibodi Hospital. Face validity was evaluated through patient interviews, using guide questions. The internal consistency was calculated using Cronbach's alpha. RESULTS: In total, 91.8% of patients declared that the ESAS-Thai questionnaire was generally clear. It yielded a Cronbach's alpha of 0.75 in the inpatient setting. After modifying the words "appetite" and "well-being," 37 cancer patients, whose mean (standard deviation) age was 52.2 (10.8) years and who were cared for by the Departments of Medicine, Surgery, Gynecology, and Otolaryngology, self-administered the questionnaire in the outpatient department. The Cronbach's alpha in the validation sample was 0.89. CONCLUSION: After the translation and cross-cultural adaptation, the Thai version of the ESAS achieved good levels of face validity and internal consistency. It is now available as a patient-administered instrument to evaluate symptoms among palliative care patients in Thailand. PMID- 21621965 TI - Epothilone-induced peripheral neuropathy: a review of current knowledge. AB - CONTEXT: Epothilones, belonging to the family of microtubule stabilizing agents, have shown prolonged remissions and improved survival in various types of refractory, treatment-resistant cancer. Ixabepilone (BMS-247550) is the main representative of these compounds. Peripheral neuropathy is a significant toxicity of epothilones, eventually resulting in dose modification and changes in the treatment plan. OBJECTIVES: This review critically looks at the pathogenesis, incidence, risk factors, characteristics, and management of epothilone-induced peripheral neuropathy (EIPN). We also highlight areas of future research to pursue. METHODS: References were identified by searches of PubMed from 2000 until December 2010 with related terms. RESULTS: The mechanism underlying EIPN remains rather unclear. Damage to the ganglion soma cells and peripheral axons through disruption of microtubules of the mitotic spindle and by interference with the axonal transport in the affected neurons may significantly contribute to the pathogenesis of EIPN. As a result, epothilones primarily produce an axonal, dose dependent, sensory distal peripheral neuropathy, which is reversible in most cases on discontinuation of treatment. The incidence of EIPN is mainly related to risk factors, including cumulative dose and probably pre-existing neuropathy. To date, apart from the use of dose reduction and schedule change algorithm, there is no effective treatment with neuroprotective agents for EIPN. CONCLUSION: EIPN remains a very challenging area in the field of toxic neuropathies. As such, there is a need for further preclinical and prospective clinical studies to elucidate the pathogenesis of EIPN and provide further robust evidence on its incidence, course, and reversibility. PMID- 21621966 TI - The therapeutic alliance in the cognitive behavioral treatment of pediatric obsessive-compulsive disorder. AB - Although cognitive-behavioral therapy (CBT) for pediatric obsessive-compulsive disorder (OCD) is considered a first-line treatment, not all youth have a positive treatment response, suggesting need for investigating factors that may enhance or reduce treatment effects. Few studies have investigated predictors of treatment response in pediatric OCD, and there is an absence of studies examining the influence of treatment process variables (e.g., therapeutic alliance [TA]) on treatment outcome. Using a multiple-informant and multiple-time point design, the current study examined the role of the TA in family-based CBT for pediatric OCD. Analyses examined (1) the predictive value of the TA on OCD symptom reduction and (2) whether changes in the TA over time predict treatment response. Findings indicated that (1) stronger child-rated, parent-rated, and therapist-rated TAs were predictive of better treatment outcome and (2) larger and more positive early alliance shifts (as rated by changes in child-rated TA between sessions 1 and 5) were predictive of better treatment outcome. Implications for the treatment of youth with OCD within family-based CBT are discussed. PMID- 21621967 TI - Free-floating thrombus in the internal carotid artery: diagnosis and treatment of 16 cases in a single center. AB - BACKGROUND: Free-floating thrombus in the internal carotid artery (FFT-ICA) is a rare condition and its real incidence is unknown. The most common etiology is a complication of an atherosclerotic plaque, but several medical conditions can be responsible. The purpose of this study was to retrospectively analyze our experience with carotid endarterectomy in the management of FFT-ICA and also to analyze the patient outcome. METHODS: A retrospective review was performed of all patients admitted during the past 9 years with a diagnosis of FFT-ICA. Patient demographics, clinical manifestations, diagnostic modalities, surgical indications, operative details, postoperative courses, and follow-up information were recorded from the hospital database. RESULTS: Between January 2000 and December 2008, in our Unit, 2,572 carotid endarterectomies were performed for carotid artery disease. A total of 16 patients (16 of 2,572; 0.62%) were treated for an FFT-ICA. In all, 87.5% (14 of 16) of patients had neurological symptoms. All patients underwent a duplex scan. In 75% (12 of 16) of cases, additional diagnostic tests were performed: digital subtraction angiography (DSA), magnetic resonance angiography, or computed tomographic scan. Duplex scan and DSA detected the FFT-ICA in 62.5% and 100% of cases, respectively. Computed tomographic scan and magnetic resonance angiography failed to provide a diagnosis in majority of the patients (33.4% and 66.7%, respectively). The presence of FFT-ICA was confirmed intraoperatively in all cases. The cumulative stroke rate after surgery was 6.3% (one of 16). Of the total number of patients discharged, 68.75% showed an improvement of neurological symptoms, 12.5% were asymptomatic, 12.5% had no changes in symptoms, and 6.25% of cases worsened. At 30-day follow-up, the survival rate was 93.7% and 75% of patients showed an improvement of neurological symptoms, 12.5% were asymptomatic, and 6.25% died. In all, 6.25% of patients were lost to follow-up. CONCLUSION: Patients with FFT-ICA are usually symptomatic and present with an acute emergency. DSA remains the gold standard diagnostic test in FFT-ICA detection. We cannot assert that early surgery is superior to temporary anticoagulation and/or delayed intervention because of the absence of a comparison group. However, our retrospective results suggest that prompt intervention seems to be a safe alternative in FFT-ICA treatment. PMID- 21621968 TI - Anastomotic pseudoaneurysm formation after bypass grafting to porcelainized peroneal artery: successful treatment by percutaneous thrombin injection. AB - We present a rare case of a distal anastomotic pseudoaneurysm after bypass grafting to a porcelainized peroneal artery. Successful management was achieved by percutaneous thrombin injection. This case illustrates the value of nonsurgical intervention for this rare but difficult problem. PMID- 21621969 TI - A true aneurysm of posterior tibial artery. AB - True aneurysms of tibial artery are uncommon. We report a case of a 47-year-old woman who suffered from a distal embolism in the left toes. The surgical intervention involved an aneurysmectomy and the interposition of the posterior tibial artery using the saphenous vein graft. She has been doing well 22 months after the operation. PMID- 21621970 TI - Spinal cord, bowel, and buttock ischemia after endovascular aneurysm repair. AB - A 66-year-old man with multiple comorbidities presented with a juxtarenal perianastomotic aortic aneurysm 10 years after open abdominal aortic aneurysm repair. The aneurysmal disease also involved both iliac bifurcations, the right internal iliac artery, the left common femoral artery (CFA) up to its bifurcation, and the homolateral popliteal artery. We performed bilateral internal iliac artery coil embolization 1-month apart. Later, we performed aortouniiliac endografting extending to the right external iliac artery and placement of an endovascular plug in the left external iliac artery. A right CFA to left femoral bifurcation bypass graft was then constructed after ligation of the left CFA aneurysm. After recovering from anesthesia and despite sequential hypogastric embolization, the patient developed postoperative paraplegia, buttock ischemia, and ischemic colitis and died on postoperative day 5. The possible pathogenic mechanisms involved in the onset of these ischemic complications are discussed in this article. PMID- 21621971 TI - Reversal of delayed-onset paraparesis after revision thoracic endovascular aortic repair for ruptured thoracic aortic aneurysm. AB - Thoracic endovascular aortic repair (TEVAR) is an important surgical option for the emergency treatment of ruptured thoracic aortic aneurysms, but is associated with a risk of spinal cord ischemia (SCI). Although risk factors for the development of SCI have been well described, the effectiveness of treatment to increase spinal cord perfusion pressure remains incompletely understood. We report the successful treatment of delayed-onset paraparesis after revision TEVAR for acute descending thoracic aortic rupture with the combined use of blood pressure augmentation and cerebrospinal fluid drainage. The clinical manifestations, pathophysiology, and management of SCI after TEVAR are reviewed. PMID- 21621972 TI - Complete removal of infected abdominal aortic stent-graft with suprarenal fixation. AB - Stent-graft infections after endovascular aneurysm repair are rare but can have devastating consequences. Open surgery to treat such infections is associated with considerable morbidity and mortality. Removal of the stent-graft is technically challenging, especially when it has a suprarenal fixation. Several in situ reconstructions have been described, with varying results. We report a case in which a Zenith stent-graft became infected after endovascular aneurysm repair to treat an abdominal aortic aneurysm and blue toe syndrome. The endoprosthesis was removed completely, and in situ reconstruction was performed successfully and without complications by using a rifampin-soaked Dacron graft. PMID- 21621973 TI - Relationship between acoustic parameters and body mass analysis in young males. AB - PURPOSE: To analyze the correlation between acoustic parameters and body height, weight, and mass composition in young males. MATERIAL: A total of 40 male subjects were included in this study. Each subject underwent acoustic analysis using the Kay Elemetric VISI Pitch (Model 3300, KayPentax, Lincoln Park, NJ) and complete body mass analysis. Pearson correlation was calculated to estimate the strength of the relationship between acoustic parameters and each of the weight analysis variables. RESULTS: The mean age of the male subjects was 24 years with a range between 18 and 40 years. The average weight and height were almost 80 and 180cm, with standard deviation (SD) of 7.42 and 10.46, respectively. The fat weight ranged between 3 and 25kg, with the main concentration being in the extremities, 61.84%+/-17.4 and less concentrated in the trunk, 16.20% +/- 7.6. The mean fundamental frequency was 120.13Hz with an SD of 19.16Hz. The mean Habitual Pitch was 114.16Hz with an SD of 16.55Hz. There was no significant correlation between the acoustic parameters and any of the body composition variables, in particular fat weight and distribution. There was a weak correlation between Shimmer, trunk fat (r value=0.328, P=0.039), and muscle mass (r value=0.326, P=0.038). CONCLUSION: The body mass composition and distribution do not correlate significantly with the fundamental frequency and the Habitual pitch. PMID- 21621974 TI - Validation of the Lithuanian version of the Glottal Function Index. AB - OBJECTIVE: To culturally adapt the Glottal Function Index (GFI) questionnaire to the Lithuanian language, and to validate it. METHODS: Psychometric analyses were performed on the translated Lithuanian version of the GFI (GFI-LT) as described by the Scientific Advisory Committee of the Medical Outcomes Trust. The GFI-LT was completed by 50 voice-disordered individuals and by 50 healthy subjects of the control group. Validity, reliability, reproducibility, sensitivity, and responsiveness to clinical change of the GFI-LT were evaluated. To assess the concurrent validity of the GFI-LT, all participants also completed the Voice Handicap Index (VHI) questionnaire. RESULTS: The GFI-LT showed a statistically significantly high reliability and internal consistency (Cronbach alpha=0.8, r=0.50), and moderate item-total correlation (r=0.41-0.55). Cronbach alpha coefficients of the test-retest reliability were above the standard (>=0.9) for individuals testing. There was a statistically significant difference between the mean scores of the control and the voice-disordered groups (P<0.001). The Receiver Operating Characteristic test indicated that the GFI-LT score of >3.0 was the optimal score distinguishing patients and healthy controls with the sensitivity of 88% and specificity of 84%. Statistically significant (P<0.05) strong correlations were found between the GFI-LT and VHI scores. The GFI-LT was found to be a responsive measurement instrument to patients' clinical statement; mean difference of the GFI-LT scores in the group of voice-disordered patients before and after surgical treatment was 5.7 (P<0.001). CONCLUSION: The GFI-LT is considered to be a valid and reliable tool for self-assessment of the severity of voice disorders in Lithuanian-speaking patients. PMID- 21621975 TI - Vocal fold vibratory characteristics in normal female speakers from high-speed digital imaging. AB - The purpose of the study was to investigate relationships between vocal fold vibrations and voice quality. Laryngeal images obtained from high-speed digital imaging (HSDI) were examined for their open-closed timing characteristics and perturbation values. A customized software delineated the glottal edges and used the Hilbert transform-based method of analysis to provide objective quantification of glottal perturbation. Overlay tracings of the transformed glottal cycles provided visual patterns on the overall vibratory dynamics. In this paper, we described the use of this method in looking at vibratory characteristics of a group of young female speakers (N=23). We found that, females with no voice complaints and who had been perceived to have normal voices were not a homogeneous group in terms of their glottal vibratory patterns during phonation. Their vibratory patterns showed characteristics similar to exemplar voices targeted to be clear (50%), pressed (27%), breathy (15%), or a mixed quality (8%). Perturbation range in terms of cycle-to-cycle frequency and amplitude was small and did not discriminate patterns. All these patterns yielded perceptually normal voices suggesting that in normal young speakers, the level of perturbation may be more important to the judgment than the actual pattern of closure. PMID- 21621976 TI - Essential rate for approximation by spherical neural networks. AB - We consider the optimal rate of approximation by single hidden feed-forward neural networks on the unit sphere. It is proved that there exists a neural network with n neurons, and an analytic, strictly increasing, sigmoidal activation function such that the deviation of a Sobolev class W2(2r)(S(d)) from the class of neural networks Phi(n)(phi), behaves asymptotically as n(-2r/d-1). Namely, we prove that the essential rate of approximation by spherical neural networks is n(-2r/d-1). PMID- 21621977 TI - Diagnostic radiograph based 3D bone reconstruction framework: application to the femur. AB - Three dimensional (3D) visualization of anatomy plays an important role in image guided orthopedic surgery and ultimately motivates minimally invasive procedures. However, direct 3D imaging modalities such as Computed Tomography (CT) are restricted to a minority of complex orthopedic procedures. Thus the diagnostics and planning of many interventions still rely on two dimensional (2D) radiographic images, where the surgeon has to mentally visualize the anatomy of interest. The purpose of this paper is to apply and validate a bi-planar 3D reconstruction methodology driven by prominent bony anatomy edges and contours identified on orthogonal radiographs. The results obtained through the proposed methodology are benchmarked against 3D CT scan data to assess the accuracy of reconstruction. The human femur has been used as the anatomy of interest throughout the paper. The novelty of this methodology is that it not only involves the outer contours of the bony anatomy in the reconstruction but also several key interior edges identifiable on radiographic images. Hence, this framework is not simply limited to long bones, but is generally applicable to a multitude of other bony anatomies as illustrated in the results section. PMID- 21621978 TI - Regulatory T-Cell (Treg) hybridoma as a novel tool to study Foxp3 regulation and Treg fate. AB - The CD25(+)Foxp3(+) regulatory T-cells (Treg) that had lost CD25 and Foxp3 in vivo (ex-Treg) exist but are difficult to study. We generated antigen (Ag) specific Treg hybridomas from iTreg clones (iTreg-hyb) using iTreg of DO11.10.Foxp3-GFP mice and presented evidence that they behave like ex-Treg. The iTreg-hyb displayed little CD25 and Foxp3-GFP but strong expression could be induced with OVA(323-339) in the presence of Ag-presenting cells, rIL-2 and rTGF beta1. They displayed all of the iTreg-associated markers examined except CTLA-4, the latter was also absent in the ex-Treg. They lacked the Helios transcription factor, suggesting they were derived from iTreg. Similar to ex-Treg, the iTreg hyb produced high level of IL-2 and Foxp3 under specific activation conditions. Two unusual properties were observed. First, the ability to induce Foxp3-GFP upon activation is progressively lost in culture over a period of 2-4 weeks. Second, Rag2(-/-) spleen cells alone selectively induced Foxp3-GFP expression albeit 30 times less efficient than Ag-specific activation. We identified cell-free supernatant, IL-6, IL-9, and IL-27 as Foxp3-inducing factors. Our study has significant implications to the stability, plasticity and fate of Treg. The usefulness and limitation of iTreg-hyb as a novel tool to study Foxp3 regulation and the fate of specific Treg subsets are discussed. PMID- 21621979 TI - Assessment of linear measurements of bone for implant sites in the presence of metallic artefacts using cone beam computed tomography and multislice computed tomography. AB - The objective was to evaluate the influence of dental metallic artefacts on implant sites using multislice and cone-beam computed tomography techniques. Ten dried human mandibles were scanned twice by each technique, with and without dental metallic artefacts. Metallic restorations were placed at the top of the alveolar ridge adjacent to the mental foramen region for the second scanning. Linear measurements (thickness and height) for each cross-section were performed by a single examiner using computer software. All mandibles were analysed at both the right and the left mental foramen regions. For the multislice technique, dental metallic artefact produced an increase of 5% in bone thickness and a reduction of 6% in bone height; no significant differences (p>0.05) were detected when comparing measurements performed with and without metallic artefacts. With respect to the cone-beam technique, dental metallic artefact produced an increase of 6% in bone thickness and a reduction of 0.68% in bone height. No significant differences (p>0.05) were observed when comparing measurements performed with and without metallic artefacts. The presence of dental metallic artefacts did not alter the linear measurements obtained with both techniques, although its presence made the location of the alveolar bone crest more difficult. PMID- 21621980 TI - Systolic blood pressure at admission, clinical manifestations, and in-hospital outcomes in patients with acute myocardial infarction. AB - BACKGROUND: Several clinical studies have demonstrated an inverse relationship between systolic blood pressure (SBP) at admission and in-hospital mortality in patients hospitalized for acute myocardial infarction (AMI). However, data on the relation between admission SBP and in-hospital prognosis in AMI patients are still lacking in Japan. METHODS AND RESULTS: A total of 1211 AMI patients were classified into quintiles based on SBP at hospital admission (<106 mmHg, n = 241; 106-125 mmHg, n = 239; 126-140 mmHg, n = 244; 141-159 mmHg, n = 238; and >= 160 mmHg, n = 249). The patients with SBP < 106 mmHg tended to have higher age, Killip class >= 3 at admission, right coronary artery, left main trunk, or multivessels as culprit lesions, larger number of diseased vessels, lower Thrombolysis In Myocardial Infarction grade in the infarct-related artery before primary percutaneous coronary intervention (PCI), and higher value of peak creatine phosphokinase concentration. Patients with SBP <106 mmHg had a significantly higher mortality, while mortality was not significantly different among the other quintiles: 25.7% (<106 mmHg), 5.4% (106-125 mmHg), 5.7% (126-140 mmHg), 2.5% (141-159 mmHg), and 5.6% (>= 160 mmHg) (p<0.001). On multivariate analysis, Killip class >= 3 at admission, admission SBP <106 mmHg, and age were the independent positive predictors of in-hospital mortality, whereas admission SBP 141-159 mmHg and primary PCI were the negative ones, but admission SBP 106 125 mmHg, admission SBP 126-140 mmHg, and admission SBP >= 160 mmHg were not. CONCLUSIONS: These results suggest that admission SBP 141-159 mmHg might be correlated with better in-hospital prognosis, whereas admission SBP <106 mmHg was associated with in-hospital death in Japanese patients hospitalized for AMI. PMID- 21621981 TI - On the clinical implications of beta-lactam-aminoglycoside synergism. PMID- 21621982 TI - Transcranial direct current stimulation for the outpatient treatment of poor responder depressed patients. AB - Transcranial direct current stimulation (tDCS) is a selective, painless, brain stimulation technique that allows the electric stimulation of specific cortical regions. TDCS has been recently used as investigational intervention for major depression and treatment resistant depression (TRD) with encouraging results. The present study was aimed to investigate the efficacy and tolerability of tDCS in major depressives with poor response to pharmacological treatment. Twenty-three depressed patients, with a diagnosis of major depressive disorder or bipolar disorder, were treated with augmentative tDCS for 5 days, two sessions per day in a blind-rater trial. The course of depressive symptoms was analyzed using repeated measures ANOVA for HAM-D and MADRS total scores. A qualitative analysis on the basis of the HAM-D response was performed as well. Both analyses were conducted at three time-points: T0 (baseline), T1 (endpoint tDCS) and T2 (end of the first week of follow-up). All patients completed the trial without relevant side-effects. A significant reduction of HAM-D and MADRS total scores was observed during the study (P<0.0001). Treatment response (endpoint HAM-D reduction >=50%) was obtained by four patients (17.4%) at T1 and by seven patients (30.4%) at T2 and remission (endpoint HAM-D<8) by three patients (13.0%) at T1 and by four subjects (17.4%) at T2. Present findings support the efficacy and good tolerability of tDCS in the acute treatment of patients with TRD with clinical benefit being progressive and extended to the first week of follow-up. Further sham-controlled trials with longer follow-up are needed to confirm present results. PMID- 21621983 TI - Span 80 vesicles have a more fluid, flexible and "wet" surface than phospholipid liposomes. AB - The surface properties of Span 80 vesicles at various cholesterol contents, together with those of various liposomes, were characterized by using fluorescence probes. The membrane fluidity of the Span 80 vesicles was measured by 1,6-diphenyl-1.3.5-hexatriene (DPH) and trimethlyammonium-DPH (TMA-DPH), and the results suggested that the surface of the Span 80 vesicles was fluid due to the lateral diffusion of Span 80 molecules. The depolarization measured by TMA DPH and the headgroup mobility measured by dielectric dispersion analysis indicated the high mobility of the head group of Span 80 vesicles. This suggested that the surface of Span 80 vesicles was flexible due to the head group structure of Span 80, sorbitol. In addition, spectrophotometric analysis with 6-dodecanoyl N, N-dimethyl-2-naphthylamine and 8-anilino-1-naphthalenesulfonic acid indicated that the water molecules could easily invade into the interior of the Span 80 vesicle membrane, suggesting that the membrane surface was more wet than the liposome surface. These surface properties indicated that the protein could interact with the interior of vesicle membranes, which was similar to the case of cholesterol. Thus the present results confirmed that the Span 80 vesicle surfaces showed the unique characteristics of fluidity, flexibility, and "wetness", whereas the liposome surfaces did not. PMID- 21621984 TI - Effect of plant-based phenol derivatives on the formation of Cu and Ag nanoparticles. AB - The complexes formed on the reaction of various metal ions viz., Cu(II) and Cu(I) with phenol derivatives viz. catechol, chlorogenic acid (CGA), hydroquinone and n propyl gallate (nPG) were established by UV-visible spectroscopy. The metal/ligand complexing ratio and complexation constants have been determined. Further, we showed that nanoparticles of Cu can be prepared from metal-phenol complexes in the presence of a protein (gelatin) by gamma-irradiation showing that the reduction is metal ion centered. Formation of Ag nanoparticles was also observed on photo-irradiation with xenon lamp in the presence of dihydroxy benzene. The Ag and Cu nanoparticles were characterized by transmission electron microscopy (TEM) and UV-visible absorption spectroscopy. TEM technique showed the presence of Cu and Ag nanoparticles with average size of 20 and 30 nm, respectively. PMID- 21621985 TI - Dependence of liquid crystal morphology on phospholipid hydrocarbon length. AB - The liquid crystal morphologies of symmetrical diacy phosphatidylcholine liposomes examined in this research study were found to be dependent on saturated hydrocarbon chain length. Both powder X-ray diffraction and synchrotron mid-IR spectromicroscopy indicate that phosphatidylcholines with short hydrocarbon tails (i.e. ten and twelve carbons) are more likely to form unilamellar liposomes while those with long hydrocarbon tails (i.e. eighteen and twenty carbons) are more likely to form multilamellar liposomes. Hydrocarbon chain lengths of fourteen and sixteen represent a transitional zone between these two liquid crystal morphologies. The FTIR spectra where a shoulder develops on the peak at wavenumber 1750 cm(-1) particularly highlights the change in the packing of adjacent molecules in the transitional zone. PMID- 21621986 TI - Efficacy of fish liver oil and propolis as neuroprotective agents in pilocarpine epileptic rats treated with valproate. AB - OBJECTIVE: To evaluate the action of fish liver oil and propolis in pilocarpine epileptic rats treated with the anticonvulsant drug valproate. METHODS: Seven groups of rats were treated daily for six months: control; fish liver oil (0.4ml/kg b.w); propolis (50mg/kg b.w); pilocarpine-treated rats (epileptic control); epileptic rats treated with valproate (400mg/kg b.w); groups 6 and 7, epileptic rats treated with valproate plus fish liver oil or propolis. RESULTS: Pilocarpine administration caused a significant increase in hippocampal dopamine and serotonin levels accompanied with a significant decrease in their levels in serum. Lipid peroxidation level and LDH activity in hippocampus were significantly increased after pilocarpine treatment whereas Na(+)/K(+)-ATPase activity and total antioxidant capacity were significantly decreased compared to the controls. Animals treated with the combined treatments showed a significant improvement in tested parameters towards the normal values of the control. CONCLUSION: Fish liver oil and propolis when given in combination with valproate, neuroprotected against the neurophysiological disorders induced by pilocarpine epilepsy in rats. PMID- 21621987 TI - [Indoor air quality in schools]. AB - Indoor air quality in schools has received particular attention over the past several years. Children are considered as one of the most sensitive groups to atmospheric pollution because their bodies are actively growing and they breathe higher volumes of air relative to their body weights than adults do. They also spend more time in school or group structures (preschools, day nurseries) than in any indoor environments other than the home. The analysis of children's exposure to air pollution at school requires the identification of the main pollutant sources present in these educational institutions. Both a strong contribution of outdoor pollution and a very specific pollution bound to school activities such as the use of paints, markers, glues, and manufactured ink eraser pens, exist. The ventilation in school buildings also plays an important role in air quality. A higher air exchange may improve thermal comfort and air quality. The cause of indoor air pollution is a combinatory effect of physical, chemical, and biological factors, and the adequacy of ventilation in the environment. Several pollutants have been reported to exist in classrooms such as bacteria, molds, volatile organic compounds, persistent organic pollutants and microparticles. There is a correlation between the concentrations of the pollutants and onset of health problems in schoolchildren. We observe predominantly respiratory symptoms as well as a prevalence of respiratory diseases such as asthma and allergies. This study shows that poor indoor air quality affects children's health. PMID- 21621988 TI - [Can we really use the Stewart-Fencl method to analyze acid-base derangements in clinical practice?]. PMID- 21621989 TI - [Persistent dysphonia showing a laryngeal foreign body in a child]. AB - INTRODUCTION: Inhalation of a laryngotracheobronchial foreign body is a common pediatric emergency situation. It is a source of morbidity and even mortality, especially among children under 3 years of age. CASE REPORT: A 14-month-old child presented suddenly combining bitonal dysphonia and dyspnea. Given the persistence of symptoms after 1.5 months and the normality of examinations requested by his doctor (pH, cervical ultrasonography, cervical and thoracic radiography), an ENT opinion was sought. An aerodigestive tract endoscopy was carried out in the emergency setting, finding a glottic foreign body associated with subglottic granulomas. The foreign body extraction led to the immediate disappearance of dyspnea. Dysphonia gradually improved under Budesonide aerosols. CONCLUSION: The lack of penetration syndrome in the interrogation and non specific symptoms may lead to an important diagnosis and treatment delay with dramatic consequences in case of airway foreign body. Endoscopy under general anesthesia must be practiced if there is any doubt for a thorough examination of the airways. PMID- 21621990 TI - Effect of surgical modality and hypofractionated split-course radiotherapy on local control and survival from sinonasal mucosal melanoma. AB - AIMS: To assess the efficacy of surgery and high-dose split-course radiotherapy in sinonasal head and neck mucosal melanoma (SHNMM). MATERIAL AND METHODS: Between 1991 and 2006, 23 patients (median age 73 years, male:female ratio 0.4) with non-metastatic SHNMM underwent surgery and postoperative radiotherapy, two had exclusive radiotherapy. Radiotherapy consisted of three series of 18Gy (3*6Gy every other day for 1 week) with 3 week planned treatment breaks. Chi-squared tests, Kaplan-Meyer method and Log-rank test were used to assess prognostic factors for survival and local control. RESULTS: There were 20 nasal cavity tumours; 12 of these involved more than one sinonasal site. One patient (4%) had lymphadenopathies at diagnosis. Six SHNMMs (24%) were amelanotic. The median follow-up was 39 months. Fourteen patients had en bloc surgery, 16 underwent radiation (14 postoperative, two exclusive). Eleven patients had local relapse, three had regional relapse and three had bone or liver metastases. Five year local control was 49+/-12%. Five year overall and SHNMM-specific survival was 38+/-12% and 62+/-12%, respectively. Five patients were alive without disease after 5 years and three after 10 years. En bloc excision (tumour removed in one piece) was prognostic for survival. CONCLUSIONS: En bloc surgery was a prognostic factor on outcomes for local control and survival in this series. Data from the literature have shown that postoperative radiation therapy improves local control. Most series were carried out with conventional fractionation. The effect of planned breaks (split-course radiotherapy) may be deleterious, as suggested in this series. Therefore, split-course radiotherapy cannot be recommended for SHNMM. PMID- 21621991 TI - Control of experimental pulmonary tuberculosis depends more on immunostimulatory leukotrienes than on the absence of immunosuppressive prostaglandins. AB - Prostaglandins (PGs) and leukotrienes (LTs) are produced in Mycobacterium tuberculosis (Mtb)-infected lungs and have immune suppressive and protective effects, respectively. Considering that both of these mediators are produced during mycobacterial infection, we investigated the specific and relative biological importance of each in regulating host response in experimental tuberculosis. Administration of celecoxib, which was found to reduce lung levels of PGE(2) and increase LTB(4), enhanced the 60-day survival of Mtb-infected mice in 14%. However administration of MK-886, which reduced levels of LTB(4) but did not enhance PGE(2), reduced 60-day survival from 86% to 43% in Mtb-infected mice, and increased lung bacterial burden. MK-886 plus celecoxib reduced survival to a lesser extent than MK-886 alone. MK-886- and MK-886 plus celecoxib-treated animals exhibited reduced levels of the protective interleukin-12 and gamma interferon. Our findings indicate that in this model, the protective effect of LTs dominates over the suppressive effect of PGs. PMID- 21621992 TI - Lung function and respiratory symptoms in a 1-year randomized smoking cessation trial of varenicline in COPD patients. AB - There are few data concerning changes in lung function and respiratory symptoms in smokers with chronic obstructive pulmonary disease (COPD) weeks to months after quitting smoking. We examined serial changes in spirometry and Clinical COPD Questionnaire (CCQ) scores (measuring respiratory symptoms and health related quality of life) in COPD participants by smoking status during a smoking cessation trial. In this randomized, double-blind trial, smokers with mild-to moderate COPD were treated with varenicline 1 mg b.i.d. or placebo for 12 weeks and followed to Week 52. Primary endpoints of abstinence were previously reported. Secondary endpoints were mean changes from baseline in post bronchodilator forced expired volume in 1 s (FEV(1)) and CCQ scores. Change from baseline in post-bronchodilator FEV(1) was significantly improved in continuous abstainers (121.8 mL) vs. continuous smokers (37.9 mL) at Week 12 (P = 0.0069), but not at Weeks 24 or 52. Mean change from baseline at Week 12 in CCQ Total Score was significantly better in continuous abstainers (-1.04) vs. continuous smokers (-0.53; P < 0.0001): this improvement was sustained at Weeks 24 and 52. In a 1-year cessation trial of smokers with COPD, continuous abstinence compared with continuous smoking significantly improved post-bronchodilator FEV(1) at Week 12 (although the difference narrowed subsequently) and CCQ Total Scores at Week 12, with sustained improvement thereafter. ( TRIAL REGISTRY: http://www.clinicaltrials.gov; trial identifier: NCT00285012). PMID- 21621993 TI - Effects of inspiratory muscle training in patients with heart failure. AB - AIM: To investigate the effects of inspiratory muscle training (IMT) on functional capacity and balance, respiratory and peripheral muscle strength, pulmonary function, dyspnea, fatigue, depression, and quality of life in heart failure patients. METHODS: A prospective, randomized controlled, double-blinded study. Thirty patients with heart failure (NYHA II-III, LVEF<40%) were included. Sixteen patients received IMT at 40% of maximal inspiratory pressure (MIP), and 14 patients received sham therapy (15% of MIP) for 6 weeks. Functional capacity and balance, respiratory muscle strength, quadriceps femoris muscle strength, pulmonary function, dyspnea, fatigue, quality of life, and depression were evaluated. RESULTS: Functional capacity and balance, respiratory and peripheral muscle strength, dyspnea, depression were significantly improved in the treatment group compared with controls; quality of life and fatigue were similarly improved within groups (p < 0.05). Functional capacity (418.59 +/- 123.32 to 478.56 +/- 131.58 m, p < 0.001), respiratory (MIP = 62.00 +/- 33.57 to 97.13 +/- 32.63 cmH(2)O, p < 0.001) and quadriceps femoris muscle strength (240.91 +/- 106.08 to 301.82 +/- 111.86 N, p < 0.001), FEV(1)%, FVC% and PEF%, functional balance (52.73 +/- 3.15 to 54.25 +/- 2.34, p < 0.001), functional dyspnea (2.27 +/- 0.88 to 1.07 +/- 0.79, p < 0.001), depression (11.47 +/- 7.50 to 3.20 +/- 4.09, p < 0.001), quality of life, fatigue (42.73 +/- 11.75 to 29.07 +/- 13.96, p < 0.001) were significantly improved in the treatment group. Respiratory muscle strength (MIP = 78.64 +/- 35.95 to 90.86 +/- 30.23 cmH(2)O, p = 0.001), FVC%, depression (14.36 +/- 9.04 to 9.50 +/- 10.42, p = 0.011), quality of life and fatigue (42.86 +/- 12.67 to 32.93 +/- 15.87, p = 0.008) were significantly improved in the control group. CONCLUSION: The IMT improves functional capacity and balance, respiratory and peripheral muscle strength; decreases depression and dyspnea perception in patients with heart failure. IMT should be included effectively in pulmonary rehabilitation programs. PMID- 21621994 TI - Enhanced methane recovery by food waste leachate injection into a landfill in Korea. AB - The current food waste leachate (FWL) disposal practice in Korea warrants urgent attention and necessary action to develop an innovative and sustainable disposal strategy, which is both environmentally friendly and economically beneficial. In this study, methane production by FWL injection into a municipal solid waste landfill with landfill gas (LFG) recovery facility was evaluated for a period of more than 4 months. With the target of recovering LFG with methane content ~50%, optimum LFG extraction rate was decided by a trial and error approach during the field investigation in five different phases. The results showed that, upon FWL injection, LFG extraction rate of ~20 m(3)/h was reasonable to recover LFG with methane content ~58%. Considering the estimated methane production potential of 31.7 m(3) CH(4) per ton of FWL, methane recovery from the landfill was enhanced by 14%. The scientific findings of this short-term investigation indicates that FWL can be injected into the existing sanitary landfills to tackle the present issue and such landfills with efficient liner and gas collection facility can be utilized as absolute and sustainable environmental infrastructures. PMID- 21621995 TI - Label-free, chemiresistor immunosensor for stress biomarker cortisol in saliva. AB - Salivary cortisol is commonly used as a bioindicator of the psychobiologic response to environmental and psychological stressors. Current analytical approaches rely on immunoassays performed at distant, centralized laboratories and involve an elaborate specimen collection-processing-transportation-storage analysis-reporting cycle. To facilitate point-of-use measurement of salivary cortisol levels, we describe the development and proof-of-concept testing of an ultrasensitive, label-free immunosensor based on a single-walled, carbon nanotube based chemiresistive transducer. Carbon nanotubes were functionalized with a cortisol analog [cortisol-3-CMO-NHS ester] and a monoclonal anti-cortisol antibody was ligated to this receptor. Addition of phosphate buffer as well as artificial saliva spiked with varying cortisol concentrations displaced the anti cortisol antibody producing corresponding decreases in the resistance/conductance of the nanotube-biomolecule hybrid. The immunosensor demonstrated an ultralow detection limit of 1 pg/ml and excellent binding selectivity for cortisol even in the presence of structurally similar steroids such as 21-hydroprogesterone. The nanotube immunosensor offers attractive prospects for the development of highly sensitive biosensor for rapid, label-free measurement of salivary cortisol in a variety of clinical and research settings. PMID- 21621996 TI - Fermentative biohydrogen production systems integration. AB - Acidogenic fermentation can be used to produce hydrogen from a range of biomass sources. The effluent from this process can be utilised in a number of biological processes enabling further recovery of energy from the biomass. In this review a number of candidate technologies are assessed including conventional methanogenic anaerobic digestion, dark fermentative hydrogen production, photo-fermentation, and bioelectrochemical systems. The principles, benefits and challenges associated with integrating these technologies are discussed, with particular emphasis on integration with fermentative hydrogen production, and the current state of integrative development is presented. The various system configurations for potential integrations presented here may simultaneously permit an increase in the conversion efficiency of biomass to energy, improved adaptability to varying operating conditions, and improved stability. Such integration, while increasing system complexity, may mean that these bioprocesses could be deployed in a wider range of scenarios and be used with a greater range of substrates. PMID- 21621997 TI - Synthesis and antiviral activity of cyclopropyl-spirocarbocyclic adenosine, (4R,5S,6R,7R)-4-(6-amino-9H-purin-9-yl)-7-(hydroxymethyl)spiro[2.4]heptane-5,6 diol against hepatitis C virus. AB - An efficient method was developed for the synthesis of 6-exocyclic methylene carbocyclic intermediate 4. The Simmons-Smith cyclopropanation protocol was applied on the 6-exocyclic methylene of intermediate 4 and demonstrated its utility for the synthesis of novel class of a spiro-carbocyclic nucleoside analog 8. The titled compound 8 demonstrated a significant antiviral activity against HCV with EC(50) values of 0.273 and 0.368 MUM in genotypes 1A and 1B, respectively. PMID- 21621998 TI - Renin inhibitors for the treatment of hypertension: design and optimization of a novel series of pyridone-substituted piperidines. AB - An SAR campaign aimed at decreasing the overall lipophilicity of renin inhibitors such as 1 is described herein. It was found that replacement of the northern appendage in 1 with an N-methyl pyridone and subsequent re-optimization of the benzyl amide handle afforded compounds with in vitro and in vivo profiles suitable for further profiling. An unexpected CV toxicity in dogs observed with compound 20 led to the employment of a time and resource sparing rodent model for in vivo screening of key compounds. This culminated in the identification of compound 31 as an optimized renin inhibitor. PMID- 21621999 TI - Peptidomimetic inhibitors of bacterial peptide deformylase. AB - A series of N-formyl hydroxylamine peptide deformylase inhibitors containing a cyclic azaamino acid moiety between the P1' and P3' substituents are presented. Selected compounds display antibacterial activity against pathogens associated with respiratory tract infections with representative compounds showing excellent MICs against Haemophilus influenzae. PMID- 21622000 TI - Immediate reconstruction using a modified thoracodorsal adipofascial cutaneous flap after partial mastectomy. AB - The treatment of early breast cancer using breast conservation therapy (BCT) usually ensures local control and acceptable cosmetic results. We report a useful technique involving the use of a thoracodorsal adipofascial cutaneous flap for reconstructing defects in the upper-outer quadrant area after partial mastectomy that has achieved excellent results. We developed this procedure as an oncoplastic technique for treating a Japanese woman with a similar defect and a relatively slim body. In this procedure, partial mastectomy is followed by raising the anterior half of the flap via the same skin incision, and the posterior half is raised via an additional incision posterior to the axillary area to produce a crescent of de-epithelialized skin. A C-shaped flap containing a crescent of de-epithelialized skin composed of subcutaneous fat and the fascia of the latissimus dorsi muscle is then rotated, gathered, and inset into the breast defect. PMID- 21622001 TI - Plasmodium berghei proteome changes in response to SSJ-183 treatment. AB - The benzo[a]phenoxazine derivative, SSJ-183 has shown excellent anti-malarial efficacy and safety. However, its mechanism of action is unclear. We investigated the effect of SSJ-183 on the rodent malarial parasite, Plasmodium berghei. We analyzed changes in protein expression in the erythrocytic cycle of P. berghei with or without 18 h of SSJ-183 treatment by two-dimensional gel electrophoresis. We confirmed results with matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight tandem mass spectrometry. After treatment, seven main proteins were significantly down-regulated, and two were up-regulated; results were reproduced in three independent tests. Some of these proteins were hypothetical parasite proteins or unnamed host products. However, three proteins were identified as a heat shock protein, a disulfide isomerase precursor, and berghepain-2 from P. berghei. All three showed reduced expression after SSJ-183 treatment. This suggested that SSJ-183 was a good anti-malarial drug candidate because it targeted parasite chaperone proteins. PMID- 21622002 TI - Synthesis, conformation and biology of naphthoxylosides. AB - Proteoglycans (PG) are polyanionic proteins consisting of a core protein substituted with carbohydrate chains, that is, glycosaminoglycans (GAG). The biosynthesis of GAG can be manipulated by simple xylosides carrying hydrophobic aglycons, which can enter the cell and initiate the biosynthesis. While the importance of the aglycon is well investigated, there is far less information on the effect of modifications in the xylose residue. We have developed a new synthetic protocol, based on acetal protection and selective benzylation, for modification of the three hydroxyl groups in xylose. Thus we have synthesized twelve analogs of 2-naphthyl beta-d-xylopyranoside (XylNap), where each hydroxyl group has been epimerized or replaced by methoxy, fluoro, or hydrogen. To gain more information about the properties of xylose, conformational studies were made on some of the analogs. It was found that the (4)C(1) conformation is highly predominant, accompanied by a nonnegligible population of the (2)S(0) conformation. However, deoxygenation at C3 results in a large portion of the (1)C(4) conformation. The GAG priming ability and proliferation activity of the twelve analogs, were investigated using a matched pair of human breast fibroblasts and human breast carcinoma cells. None of the analogs initiated the biosynthesis of GAG, but an inhibitory effect on endogenous PG production was observed for analogs fluorinated or deoxygenated at C4. From our data it seems reasonable that all three hydroxyl groups in XylNap are essential for the priming of GAG chains and for selective toxicity for tumor cells. PMID- 21622003 TI - Synthesis and evaluation of benzofuran-2-yl(phenyl)methanone derivatives as ligands for beta-amyloid plaques. AB - A series of benzofuran-2-yl(phenyl)methanone derivatives were synthesized and evaluated as novel probes for beta-amyloid plaques. These derivatives were produced by a Rap-Stoermer condensation reaction. Compounds with a N,N dimethylamino group displayed high affinity for Abeta(1-42) aggregates with K(i) values in the nanomolar range. Autoradiography with brain sections of AD model mice (APP/PS1) revealed that a radioiodinated probe, [(125)I]10, labeled beta amyloid plaques selectively and displayed good brain uptake (3.53% ID/g) at 2 min. The results suggest that benzofuran-2-yl(phenyl)methanone derivatives should be investigated further as potential probes for detecting beta-amyloid plaques in the AD brain. PMID- 21622004 TI - Early outcomes of surgery for oesophageal cancer in a thoracic regional unit. Can we maintain training without compromising results? AB - OBJECTIVES: Meaningful exposure to oesophageal cancer surgery during general thoracic surgical training is restricted to few centres in the United Kingdom. Our Regional Tertiary Unit remains a rare 'large-volume' oesophagectomy centre. We aimed to determine the proportion of patients operated by trainees and their perioperative outcomes. METHODS: From January 2004 to September 2009, 323 patients (229 male and 94 female, median age of 69 (range 40-92) years) underwent oesophagectomy for carcinoma in our Thoracic Surgical Unit. Data were complete and obtained from a prospective departmental database. The preoperative characteristics, operative data and postoperative results were compared between the 120 patients (37%) operated by a trainee (group T) and the remainder 203 patients operated by a consultant (group C). RESULTS: The overall incidence of mortality, anastomotic leak and chylothorax were 6.5%, 5.3% and 2.2%, respectively. There were no differences in terms of age, gender, tumour location, tumour staging, preoperative spirometry or use of neoadjuvant chemotherapy between the two groups. There was no significant difference between the consultant group and the trainee group in the following key outcome measures: postoperative mortality (8% vs 4%), incidence of respiratory complications (30% vs 25%), hospital stay (14 days vs 13 days) and number of lymph nodes excised (median of 16 vs 14). CONCLUSIONS: Training in oesophageal cancer surgery can be provided in a large-volume thoracic surgical unit. It does not seem to compromise outcomes or use of resources. PMID- 21622005 TI - Red and blue pulse timing control for pulse width modulation light dimming of light emitting diodes for plant cultivation. AB - A pulse width modulation (PWM) light dimming system containing red and blue light emitting diodes was designed and constructed. Cultivation of the plant Arabidopsis thaliana under various light dimming wave patterns was compared. Control of the pulse timing (phase of wave pattern) between red and blue light in PWM light dimming was examined. Different plant growth was obtained by changing the phase of red and blue pulses. Pulse timing control of PWM light dimming for plant cultivation has the potential to act as a method for probing photosynthesis. PMID- 21622006 TI - Complications in dentoalveolar surgery. PMID- 21622007 TI - Complications in bone grafting. AB - Autogenous bone grafts continue to have wide use for reconstructing alveolar defects because of the many advantages associated with them. Although complications are low, the harvesting of bone grafts does have the risk of morbidity, which varies based on the harvest site chosen. Patients should be informed of possible complications associated with bone harvest as well as complications that many develop at the grafted site. PMID- 21622008 TI - Methodology of electromyographic analysis of the trunk muscles during walking in healthy subjects: a literature review. AB - PURPOSE: To review and discuss the literature about the use of trunk muscle electromyography - including the use of surface or fine-wire electrodes, site of application and muscle selection - during gait analysis in healthy subjects. METHODS: The databases Pubmed, Web of Knowledge and Cochrane Library were searched. Articles were included when EMG activity of at least one trunk muscle was measured in healthy subjects during walking. RESULTS: In the 33 selected articles 491 healthy subjects walked with different velocities on a treadmill and/or overground. The activity of the M. erector spinae, M. multifidus, M. obliquus externus and internus, M. rectus abdominus, M. trapezius, M. latissimus dorsi, M. transversus abdominus, M. iliopsoas and M. quadrates lumborum was measured. Twenty-nine studies used surface electrodes, one study fine-wire electrodes, and the other three studies used a combination. There is no consensus on the exact placement site of the electrodes. CONCLUSION: Surface electrodes were used more often than fine-wire electrodes and the descriptions of the electrode locations were mostly vague and not consistent among the different studies. There is need for further research to make specific recommendations about the type of electrodes in combination with the optimal locations of application of these electrodes. PMID- 21622009 TI - The usefulness of submaximal exercise gas exchange to define pulmonary arterial hypertension. AB - BACKGROUND: The 6-minute walk test is widely used to characterize activity tolerance and response to therapy in pulmonary arterial hypertension (PAH) but provides little information about cardiopulmonary pathophysiology. The aim of the present study was to determine whether measures of pulmonary gas exchange during relatively light exercise could differentiate between PAH patients and healthy individuals and also stratify disease severity. METHODS: The study comprised 40 PAH patients and 25 matched controls. Each completed a sub-maximal exercise test, consisting of 2 minutes of rest, 3 minutes of exercise, and 1 minute of recovery. Ventilation, pulmonary gas exchange, arterial oxygen saturation (Sao(2)), and heart rate were measured throughout using a simplified gas analysis system. RESULTS: A number of gas exchange variables differentiated PAH patients and controls. End-tidal CO(2) (P(ET)co(2)) and Sao(2) were lower in PAH vs controls (31 +/- 7 vs 39 +/- 3 mm Hg and 89% +/- 5% vs 95% +/- 2%, respectively, p < 0.05). Breathing efficiency (V(E)/Vco(2) ratio) was poorer in PAH vs controls (42 +/- 10 vs 33 +/- 5, p < 0.05). In addition, P(ET)co(2) and V(E)/Vco(2) discriminated between different severities of PAH. CONCLUSIONS: Gas exchange variables obtained during light sub-maximal exercise differentiated PAH patients from healthy controls and also between different severities of PAH. Sub-maximal exercise gas exchange may be a useful end point measure in a PAH population. PMID- 21622010 TI - Self-consciousness and concepts. PMID- 21622011 TI - Reliability of superficial surgical site infections as a hospital quality measure. AB - BACKGROUND: Although rates of superficial surgical site infection (SSI) are increasingly used as measures of hospital quality, the statistical reliability of using SSI rates in this context is uncertain. We used the American College of Surgeons National Surgical Quality Improvement Program data to determine the reliability of SSI rates as a measure of hospital performance and to evaluate the effect of hospital caseload on reliability. STUDY DESIGN: We examined all patients who underwent colon resection in hospitals participating in the American College of Surgeons National Surgical Quality Improvement Program in 2007 (n = 18,455 patients, n = 181 hospitals). We first calculated the number of cases and the risk-adjusted rate of SSI at each hospital. We then used hierarchical modeling to estimate the reliability of this quality measure for each hospital. Finally, we quantified the proportion of hospital-level variation in SSI rates due to patient characteristics and measurement noise. RESULTS: The average number of colon resections per hospital was 102 (SD 65). The risk-adjusted rate of superficial SSI was 10.5%, but varied from 0 to 30% across hospitals. Approximately 35% of the variation in SSI rates was explained by noise, 7% could be attributed to patient characteristics, and the remaining 58% represented true differences in SSI rates. Just more than half of the hospitals (54%) had a reliability >0.70, which is considered a minimum acceptable level. To achieve this level of reliability, 94 cases were required. CONCLUSIONS: SSI rates are a reliable measure of hospital quality when an adequate number of cases have been reported. For hospitals with inadequate caseloads, the National Surgical Quality Improvement Program sampling strategy could be altered to provide enough cases to ensure reliability. PMID- 21622012 TI - Are targeted preoperative risk prediction tools more powerful? A test of models for emergency colon surgery in the very elderly. AB - BACKGROUND: Whether preoperative risk prediction improves with the use of more patient- and procedure-targeted models is unclear. We created a customized preoperative mortality risk prediction score for patients 80 years or older needing an emergency colectomy and compare it with existing, more generic risk assessment methods. STUDY DESIGN: A targeted mortality prediction model was created using 2007 to 2008 American College of Surgeons National Surgical Quality Improvement Program (ACS NSQIP) data and was validated using 2005 to 2006 data. We constructed a scoring system from the significant predictors identified. The model fit of our targeted score was compared with the American Society of Anesthesiologist's (ASA) score, the Surgical Risk Scale, and the ACS Colorectal Surgery Risk Calculator. RESULTS: Analyses identified 1,358 and 372 emergency colectomies in the training and validation samples, respectively. Our targeted risk prediction score had a goodness-of-fit p value greater than 0.05 (indicating a good fit) and a c-statistic of 0.77, which represents a significantly better fit compared with the ASA score, the Surgical Risk Scale, and the ACS Colorectal Surgery Risk Calculator c-statistics (0.66, 0.66, and 0.71, respectively). When using the scores to predict mortality with 80% specificity, our targeted risk prediction score was 25% more likely to predict correctly than the ACS Colorectal Surgery Risk Calculator and 33% more likely to predict correctly compared with the ASA score and Surgical Risk Scale. CONCLUSIONS: Our study presents a validated preoperative mortality score for very elderly patients needing an emergency colectomy. The greater discriminating power of this targeted score indicates that preoperative risk assessment may need to be customized to specific procedures and patient circumstances. PMID- 21622013 TI - Vascular complications of Q-fever infections. AB - INTRODUCTION: Coxiella burnetii is a strict intracellular pathogen causing Q fever, a worldwide zoonosis with an extensive animal reservoir. Chronic Q fever infections are frequently associated with cardiovascular complications, mainly endocarditis, and also aortic aneurysms and vascular-graft infection. We present four cases of chronic Q fever infections and associated vascular complications, and review the literature to identify major symptoms and assess the prevalence, treatment and outcome in these challenging patients. MATERIALS AND METHODS: The demographic and clinical data of four patients presenting at our unit were analysed. PubMed was searched to identify articles describing patients with chronic Q-fever-associated vascular complications. RESULTS: Combining our own with the published experience, 58 cases (49 male) of chronic Q-fever-associated vascular complications were identified. The average age of the patients was 64 years (range: 30-83 years). As many as 26 patients had vascular graft infections (25 Dacron/polytetrafluoroethylene (PTFE), one homograft) and 32 had infected aneurysms. The majority of these patients presented with fever (n = 40) and/or pain (n = 43). Weight loss and fatigue were seen in 25 and 14 patients, respectively. Aneurysm rupture, aorto-enteric fistulae and lower-limb embolisation were seen in nine, four and four patients, respectively. Concurrent endocarditis was seen in two patients, whereas, for 15 cases, this information was not available. Patients were treated with antibiotics for an average of 23 months (range 1-54 months). Treatment of infected vascular segments was described in 50 patients. Ten patients were treated conservatively whilst 40 underwent resection of the infected vessel and reconstruction with a graft. Major surgical complications (graft infection, n = 3;aorto-enteric fistula, n = 2; bleeding, n = 1; anastomotic leakage, n = 1; aortic dissection, n = 1; vertebral osteomyelitis, n = 3; graft thrombosis, n = 1; renal failure, n = 2; and pneumonia, n = 1) were reported in 11 cases (21%) and were not specified in 13. The overall mortality was 24% (14/58). Seven (18%) surgically treated patients died. Six of them died within 6 months of surgery and one patient at 3 years' follow-up. Seven out of 10 of the conservatively treated patients died within 3 years of diagnosis. CONCLUSION: Aneurysms associated with Q-fever infections tend to be complicated, requiring challenging surgical corrections, and long-term antibiotic treatment. Major complications and mortality rates are significant, especially in conservatively treated patients. PMID- 21622014 TI - Osteonecrosis of the jaw in oncology patients treated with bisphosphonates: prospective experience of a dental oncology referral center. AB - OBJECTIVES: The objectives of this study were to define the incidence, pain, and healing in cancer patients treated with intravenous bisphosphonates. STUDY DESIGN: The study included long-term follow-up of 99 bisphosphonate-using patients (group A) and conservative treatment of 67 patients with bisphosphonate related osteonecrosis of the jaw (BRONJ, group B) using 3 antibiotic schemes and oral hygiene. RESULTS: The frequency of zoledronic acid single-agent use was 85.9% and 69.8% in group A and B, respectively. Median follow-up was 13 months (group A) and 16 months (group B). Two patients in group A developed BRONJ (2%). Of those with BRONJ in group B who completed follow-up, healing occurred in 14.9% (7/47) and pain subsided in 80.9% (38/47). Healing was significant in patients who received pamidronate followed by zoledronic acid (P = .023) and with BRONJ stages 0 and stage I (P = .003). CONCLUSIONS: This case series suggests that oral hygiene and conservative antibiotic therapy play a role in healing and pain alleviation in BRONJ. Oral hygiene and follow-up may decrease incidence of BRONJ. PMID- 21622016 TI - Improved synthesis of 2,2'-arylmethylene bis(3-hydroxy-5,5-dimethyl-2-cyclohexene 1-one) derivatives catalyzed by urea under ultrasound. AB - Synthesis of 2,2'-arylmethylene bis(3-hydroxy-5,5-dimethyl-2-cyclohexene-1-one) derivatives catalyzed by urea via the condensation of aromatic aldehydes and 5,5 dimethyl-1,3-cyclohexanedione was carried out in 80-98% yields at 50 degrees C in aqueous media under ultrasound. This method provides several advantages such as environment friendliness, high yields and simple work-up procedure. PMID- 21622017 TI - Development of a stiffness measurement accessory for ultrasound in breast cancer diagnosis. AB - When ultrasound imaging is used for breast cancer diagnosis, the lesion's morphology is usually used to determine if the lesion is benign or malignant. Sometimes, the information provided by the procedure may not be adequate to make an accurate judgment. Additional information is needed, such as the stiffness of the lesion relative to its surrounding tissue. This paper presents an ultrasound accessory device designed to achieve this purpose. The device is easy to operate and is similar in use to a normal clinical breast ultrasound examination. A sonologist must only attach an ultrasound probe to the device and then slide it across the lesion maintaining a constant compression depth. The built-in inverse biomechanical model of the device will then calculate the predicted stiffness ratio of the lesion relative to its surrounding tissue based on the measured palpation data. Modelling and experiments have been performed on phantoms with embedded inclusions. The experimental results show that the stiffness ratio of the inclusion to its surrounding material can be accurately predicted by the handheld device. A preliminary clinical test was also performed to demonstrate the use of this device in vivo, to offer additional information to aid classification of the tumor as benign or malignant when complemented with ultrasound images. PMID- 21622018 TI - A mathematical model for creep, relaxation and strain stiffening in parallel fibered collagenous tissues. AB - A simple model is presented for the description of relaxation, creep, and strain stiffening phenomena that are observed in parallel-fibered collagenous tissues such as ligaments and tendons. In the model formulation, the tissues are assumed to be composed of collagen fibers aligned along their physiological loading direction. The collagen fibers are gradually recruited under strain and are arranged in parallel with a Maxwell element which accounts for the viscoelasticity of the proteoglycan-rich matrix. Once straight, the collagen fibers are assumed to behave as linear elastic springs. Experimental data published by Hingorani et al. [1] are used to estimate the five model parameters by fitting relaxation and strain stiffening data and the predictions are evaluated by using creep data. The influence of each parameter on describing relaxation, creep, and strain stiffening is presented. The modeling results demonstrate that, by considering the fibers' recruitment and assuming that the matrix is linear viscoelastic, a conceptually simple model can describe relaxation, creep, and strain stiffening phenomena in ligaments and tendons. PMID- 21622019 TI - Adenine nucleotide transport in plants: much more than a mitochondrial issue. AB - Adenine nucleotides play a vital role in plant metabolism and physiology, essentially representing the major energy currency of the cell. Heterotrophic cells regenerate most of the ATP in mitochondria, whereas autotrophic cells also possess chloroplasts, representing a second powerhouse for ATP regeneration. Even though the synthesis of these nucleotides is restricted to a few locations, their use is nearly ubiquitous across the cell and thereby highly efficient systems are required to transport these molecules into and out of different compartments. Here, we discuss the location, biochemical characterization and evolution of corresponding transport systems in plants. We include recent scientific findings concerning organellar transporters from plants and algae and also focus on the physiological importance of adenine nucleotide exchange in these cells. PMID- 21622020 TI - Antitubercular and fluorescence studies of copper(II) complexes with quinolone family member, ciprofloxacin. AB - Four new mixed-ligand complexes of Cu(II) with ciprofloxacin (Cip) and uninegative bidentate ligands have been synthesized and characterized. The structure of mixed-ligand complexes was investigated using spectroscopic method, physicochemical and elemental analyses. The fluorescence spectra of complexes show red shift, which may be due to the chelation by the ligands to the metal ion. It enhances ligand ability to accept electrons and decreases the electron transition energy. Antimycobacterial screening of ligand and its copper compound against Mycobacterium tuberculosis shows clear enhancement in the antitubercular activity upon copper complexation. PMID- 21622021 TI - Raman and SERS recognition of beta-carotene and haemoglobin fingerprints in human whole blood. AB - The present work reports on Raman and Surface Enhanced Raman Scattering (SERS) vibrational fingerprints of beta-carotene and haemoglobin in fresh whole blood (i.e. right after blood test) with different laser excitations, i.e. visible (514 nm) and near-infrared (NIR, 785 nm). The use of colloidal silver nanoparticles significantly increases the Raman signal, thus providing a clear SERS spectrum of blood. The collected spectra have been examined and marker bands of beta-carotene and of the haem prosthetic group of haemoglobin have been found. In particular, the fundamental features of beta-carotene (514 nm excitation), blood proteins and haem molecules (785 nm excitation) were recognized and assigned. Moreover haemoglobin SERS signals can be identified and related with its oxygenation state (oxy-haemoglobin). The data reported show the prospects of Raman and SERS techniques to detect important bio-molecules in a whole blood sample with no pre treatment. PMID- 21622022 TI - Barriers to acceptance of personal digital assistants for HIV/AIDS data collection in Angola. AB - PURPOSE: Handheld computers have potential to improve HIV/AIDS programs in healthcare settings in low-resource countries, by improving the speed and accuracy of collecting data. However, the acceptability of the technology (i.e., user attitude and reaction) is critical for its successful implementation. Acceptability is particularly critical for HIV/AIDS behavioral data, as it depends on respondents giving accurate information about a highly sensitive topic - sexual behavior. METHODS: To explore the acceptability of handheld computers for HIV/AIDS data collection and to identify potential barriers to acceptance, five focus groups of 8-10 participants each were conducted in Luanda, Angola. Facilitators presented Palm Tungsten E handhelds to the focus groups, probed participants' perceptions of the handheld computer, and asked how they felt about disclosing intimate sexual behavior to an interviewer using a handheld computer. Discussions were conducted in Portuguese, the official language of Angola, and audio-taped. They were then transcribed and translated into English for analysis. RESULTS: In total, 49 people participated in the focus groups. PDAs were understood through the lens of social and cultural beliefs. Themes that emerged were suspicion of outsiders, concern with longevity, views on progress and development, and concern about social status. CONCLUSIONS: The findings from this study suggest that personal and cultural beliefs influence participant acceptance of PDAs in Angola. While PDAs provide great advantages in terms of speed and efficiency of data collection, these barriers, if left unaddressed, may lead to biased reporting of HIV/AIDS risk data. An understanding of the barriers and why they are relevant in Angola may help researchers and practitioners to reduce the impact of these barriers on HIV/AIDS data collection. PMID- 21622023 TI - HIV-1 genetic diversity and drug resistance among treatment naive patients from Southern Brazil: an association of HIV-1 subtypes with exposure categories. AB - BACKGROUND: The AIDS epidemic in Southern Brazil has unique features, showing co circulation of HIV-1 subtypes C, B and recombinant forms. Florianopolis has the second highest AIDS incidence among Brazilian capitals, but limited information is available about HIV molecular epidemiology and prevalence of primary drug resistance. OBJECTIVES: To investigate the molecular epidemiology of HIV-1 in Florianopolis and to describe the prevalence of primary HIV-1 drug resistance mutations (DMRs). STUDY DESIGN: Epidemiological and clinical data from 82 untreated patients from Florianopolis (2008-2009) were analyzed. The HIV-1 subtype at envelope, protease, reverse transcriptase and integrase regions were determined by phylogenetic and bootscaning analyses and the drug resistance profile were analyzed at the Stanford HIV Drug Resistance Database. RESULTS: The most frequent HIV-1 genetic form was subtype C (65.8%) followed by mosaics BC (18.3%), subtype B (13.4%), subtype F1 (1.2%) and BCF1 recombinant (1.2%). HIV-1 subtype C and BC recombinants were much more frequent in the heterosexual exposure category, whereas subtype B was more common in the MSM exposure category. DRMs were seen in 11% of the sequences, 2.4% of them were related to PI, 5% to NRTI, 3.6% to NNRTI and 1.2% was related to INTI. CONCLUSIONS: The present study confirms the high prevalence of subtype C and BC recombinants in Santa Catarina State and revealed a significant difference in the subtype distribution among distinct virus exposure categories. This study also shows a relative high prevalence of protease/reverse transcriptase primary drug resistance mutations and corroborates the usefulness of the integrase inhibitors in southern Brazil. PMID- 21622024 TI - Correlates of pedometer-measured and self-reported physical activity among young Australian adults. AB - Accurately quantifying physical activity is important for investigating relations with potential correlates, but past studies have mostly relied on self-report measures, which may be susceptible to error and biases, limiting interpretability. This study aimed to examine correlates of pedometer-determined physical activity and compare them with correlates of self-reported physical activity. Cross-sectional data were taken from 2017 Australian adults (aged 26-36 years) who were involved in the Childhood Determinants of Adult Health follow-up study during 2004-2006. Daily steps were recorded for seven days using Yamax pedometers and physical activity (total min/week) was reported via the long International Physical Activity Questionnaire. Demographic, biological, behavioral, psychological, social and environmental factors were assessed. Lower education, blue collar occupation and higher mental health score (men) and low moderate alcohol intake (women) were positively associated with self-report and pedometer-measured activity. Among men, body mass index (BMI) was inversely and physical health score was positively associated with pedometer-measured activity while smoking, low to moderate alcohol intake, higher general health and urban area of residence were positively associated with self-reported activity. Among women, age and general health status were positively associated and number of live births inversely associated with pedometer-measured activity, while lower education, blue collar occupation, part time employment, smoking, diet, higher physical health score and higher mental health score were positively associated and white collar occupation inversely associated with self-reported activity. Many physical activity correlates differed depending on the measure employed; researchers should be mindful of these differences when selecting measures of physical activity. PMID- 21622025 TI - Advancing affective attributes and empowering undergraduate students--lessons learned from the Bali bombing. AB - Caring as an integral component in the nursing curriculum is enjoying a resurgence in the literature of late. The argument is that nursing education has tended to overemphasise the cognitive domain and under emphasise the affective. An alternative is to use the combined effect of cognition, imagination, intuition and emotion. This is supported by the theory of transformational learning, whereby students clarify their personal and professional purpose in life and are empowered to become informed, self-efficacious practitioners and autonomous thinkers as they negotiate personal values and meaning. In order to integrate these important theoretical concepts into everyday practice, educators need practical examples and case studies that show how caring is taught. This paper continues the conversation on narrative and transformational learning pedagogies and illustrates how affective attributes are developed through a single lecture. The aim of the lecture was to sensitise students to the human impact of terrorism and violence and the effects on both health care workers and the survivors of trauma. The rationale was that by allowing students to critically reflect on their own core knowledge and skills, they could question prior perceptions of their role, resulting in a revised or new perspective of those experiences and strengthen their belief in their abilities to cope in crisis situations. This transformative approach involved the delivery of knowledge and theory underpinning disaster response, personal narratives about a critical learning event that embodied clinically relevant lessons, activities that promoted critical self-reflection to strengthen students' beliefs in their own ability to cope by converting core knowledge into action and, finally student evaluation of the lesson (see Table 1). PMID- 21622026 TI - Simian immunodeficiency virus infection in rhesus macaques induces selective tissue specific B cell defects in double positive CD21+CD27+ memory B cells. AB - B cell dysfunction represents a central feature in HIV infection and pathogenesis. Our recent studies have shown that peripheral and lymphoid double positive CD21+CD27+ B cells were able to become activated and proliferate at higher rates than other B cell subpopulations. Increased proliferation of tonsillar memory B cells was identified compared to other tissues examined. Here, we demonstrate the decreased proliferation of tonsillar memory (CD21+CD27+) B cells during acute SIV infection also suggests that these cells may play an important role in SIV pathogenesis. Our findings demonstrate that SIV infection may induce selective defective responses in specific tissues, by suppressing memory B cell proliferation in tissues. PMID- 21622027 TI - Antiepileptic drug therapy: does mechanism of action matter? PMID- 21622028 TI - Supervised exercise, spinal manipulation, and home exercise for chronic low back pain: a randomized clinical trial. AB - BACKGROUND CONTEXT: Several conservative therapies have been shown to be beneficial in the treatment of chronic low back pain (CLBP), including different forms of exercise and spinal manipulative therapy (SMT). The efficacy of less time-consuming and less costly self-care interventions, for example, home exercise, remains inconclusive in CLBP populations. PURPOSE: The purpose of this study was to assess the relative efficacy of supervised exercise, spinal manipulation, and home exercise for the treatment of CLBP. STUDY DESIGN/SETTING: An observer-blinded and mixed-method randomized clinical trial conducted in a university research clinic in Bloomington, MN, USA. PATIENT SAMPLE: Individuals, 18 to 65 years of age, who had a primary complaint of mechanical LBP of at least 6-week duration with or without radiating pain to the lower extremity were included in this trial. OUTCOME MEASURES: Patient-rated outcomes were pain, disability, general health status, medication use, global improvement, and satisfaction. Trunk muscle endurance and strength were assessed by blinded examiners, and qualitative interviews were performed at the end of the 12-week treatment phase. METHODS: This prospective randomized clinical trial examined the short- (12 weeks) and long-term (52 weeks) relative efficacy of high-dose, supervised low-tech trunk exercise, chiropractic SMT, and a short course of home exercise and self-care advice for the treatment of LBP of at least 6-week duration. The study was approved by local institutional review boards. RESULTS: A total of 301 individuals were included in this trial. For all three treatment groups, outcomes improved during the 12 weeks of treatment. Those who received supervised trunk exercise were most satisfied with care and experienced the greatest gains in trunk muscle endurance and strength, but they did not significantly differ from those receiving chiropractic spinal manipulation or home exercise in terms of pain and other patient-rated individual outcomes, in both the short- and long-term. CONCLUSIONS: For CLBP, supervised exercise was significantly better than chiropractic spinal manipulation and home exercise in terms of satisfaction with treatment and trunk muscle endurance and strength. Although the short- and long-term differences between groups in patient-rated pain, disability, improvement, general health status, and medication use consistently favored the supervised exercise group, the differences were relatively small and not statistically significant for these individual outcomes. PMID- 21622029 TI - Adenomyosis: review of the literature. AB - Adenomyosis usually occurs in women in their reproductive years, predominantly in those with menorrhagia and dysmenorrhea. The etiology and pathophysiology remain unclear; however, recent advancements in diagnostic methods and new investigations of treatment options have changed how clinicians manage adenomyosis. A review was performed using PubMed and cross-references of reviews, case reports, and prospective and retrospective studies published from 1958 to 2010 to provide an overview of the etiology, diagnosis, prevalence, risk factors, clinical signs and symptoms, and treatments of adenomyosis. PMID- 21622030 TI - MDC1 is ubiquitylated on its tandem BRCT domain and directly binds RAP80 in a UBC13-dependent manner. AB - The cellular response to DNA damage is essential for maintenance of genomic stability. MDC1 is a key member of the DNA damage response. It is an adaptor protein that binds and recruits proteins to sites of DNA damage, a crucial step for a proper response. MDC1 contains several protein-protein interacting modules, including a tandem BRCT domain that mediates various interactions involving MDC1. Here we demonstrate that MDC1 binds directly to RAP80, which is a DNA damage response protein that recruits BRCA1 to sites of damage. The interaction between MDC1 and RAP80 requires the tandem BRCT domain of MDC1 and the ubiquitin interacting motifs of RAP80. Moreover, the interaction depends on UBC13, an E2 ubiquitin ligase that catalyzes K63-linked poly-ubiquitin chain formation. The results highly propose that the interaction between MDC1 and RAP80 depends on a ubiquitylation event, which we found to take place on K-1977 of MDC1. This study provides the first evidence that interactions involving MDC1 can be regulated by ubiquitylation. PMID- 21622031 TI - DNA damage response and transcription. AB - A network of DNA damage surveillance systems is triggered by sensing of DNA lesions and the initiation of a signal transduction cascade that activates genome protection pathways including nucleotide excision repair (NER). NER operates through coordinated assembly of repair factors into pre- and post-incision complexes. Recent work identifies RPA as a key regulator of the transition from dual incision to repair-synthesis in UV-irradiated non-cycling cells, thereby averting the generation of unprocessed repair intermediates. These intermediates could lead to recombinogenic events and trigger a persistent ATR-dependent checkpoint signaling. It is now evident that DNA damage signaling is not limited to NER proficient cells. ATR-dependent checkpoint activation also occurs in UV exposed non-cycling repair deficient cells coinciding with the formation of endonuclease APE1-mediated DNA strand breaks. In addition, the encounter of elongating RNA polymerase II (RNAPIIo) with DNA damage lesions and its persistent stalling provides a strong DNA damage signaling leading to cell cycle arrest, apoptosis and increased mutagenesis. The mechanism underlying the strong and strand specific induction of UV-induced mutations in NER deficient cells has been recently resolved by the finding that gene transcription itself increases UV induced mutagenesis in a strand specific manner via increased deamination of cytosines. The cell removes the RNAPIIo-blocking DNA lesions by transcription coupled repair (TC-NER) without displacement of the DNA damage stalled RNAPIIo. Deficiency in TC-NER associates with mutations in the CSA and CSB genes giving rise to the rare human disorder Cockayne syndrome (CS). CSB functions as a repair coupling factor to attract NER proteins, chromatin remodelers and the CSA-E3 ubiquitin ligase complex to the stalled RNAPIIo; CSA is dispensable for attraction of NER proteins, yet in cooperation with CSB is required to recruit XAB2, the nucleosomal binding protein HMGN1 and TFIIS. The molecular mechanisms by which these proteins bring about efficient TC-NER and trigger signaling after transcription arrest remain elusive; particularly the role of chromatin remodeling in TC-NER needs to be clarified in the context of anticipated structural changes that allow repair and transcription restart. PMID- 21622032 TI - Lesion bypass by S. cerevisiae Pol zeta alone. AB - DNA polymerase zeta (Pol zeta) participates in translesion synthesis (TLS) of DNA adducts that stall replication fork progression. Previous studies have led to the suggestion that the primary role of Pol zeta in TLS is to extend primers created when another DNA polymerase inserts nucleotides opposite lesions. Here we test the non-exclusive possibility that Pol zeta can sometimes perform TLS in the absence of any other polymerase. To do so, we quantified the efficiency with which S. cerevisiae Pol zeta bypasses abasic sites, cis-syn cyclobutane pyrimidine dimers and (6-4) photoproducts. In reactions containing dNTP concentrations that mimic those induced by DNA damage, a Pol zeta derivative with phenylalanine substituted for leucine 979 at the polymerase active site bypasses all three lesions at efficiencies between 27 and 73%. Wild-type Pol zeta also bypasses these lesions, with efficiencies that are lower and depend on the sequence context in which the lesion resides. The results are consistent with the hypothesis that, in addition to extending aberrant termini created by other DNA polymerases, Pol zeta has the potential to be the sole DNA polymerase involved in TLS. PMID- 21622033 TI - Surface energetics of bone mineral and synthetic hydroxyapatite using inverse gas chromatography. AB - Surface energy is one of the important factors that govern protein adhesion and cell attachment on biomaterial surfaces. Inverse gas chromatography (IGC) provides an excellent method to measure the surface energetics of rough and porous biosurfaces. In this study IGC was used to characterize and compare the surface energetics of synthetic and biological hydroxyapatites (natural bone mineral). IGC experiments were performed on three samples: synthetic hydroxyapatites with two levels of purity (99% and 90%) and natural biological hydroxyapatite obtained from bovine trabecular bone. The Lifshitz-Van der Waals component of the surface free energy (gamma(S)(LW)) and specific interaction parameter (E(pi)) were determined by using homologous series of n-alkanes and alkenes as IGC probe molecules, respectively. The synthetic hydroxyapatite had values of gamma(S)(LW) of 33.4 mJm-2 at 99% purity and 53.3 mJm-2 at 90% purity. Biological hydroxyapatite had a value of gamma(S)(LW) of 45.7 mJm-2. For the synthetic hydroxyapatite, the values of pi-bond specific interaction parameters, E(pi), were 0.95 mJ (99%) and 3.01 mJ (90%). The biological hydroxyapatite sample had a value of 2.44 mJ for E(pi). The results suggest that, as compared to the synthetic compounds, the biological apatite has considerable surface heterogeneity, either chemical (impurities) or structural suggesting a scaffold surface that is more conducive of protein adhesion and cell attachment. PMID- 21622034 TI - Liquid chromatography-tandem mass spectrometric assay for the PARP-1 inhibitor olaparib in combination with the nitrogen mustard melphalan in human plasma. AB - A bioanalytical assay for the new poly(ADP-ribose) polymerase-1 inhibitor olaparib in combination with melphalan was developed and validated. For the quantitative assay, human plasma samples were pre-treated on ice using protein precipitation with 2% (v/v) acetic acid in acetonitrile containing erlotinib and melphalan-d8 as internal standards. The extract was diluted with water and injected into the chromatographic system. This system consisted of a sub-2 MUm particle, trifunctional bonded octadecyl silica column with an isocratic elution using 0.01% (v/v) of formic acid in a mixture of water and methanol. The eluate was transferred into the electrospray interface with positive ionization and the analyte was detected in the selected reaction monitoring mode of a triple quadrupole mass spectrometer. The assay was validated in a 10-5000 ng/ml calibration range for both drugs. The lowest level of this range corresponded to the lower limit of quantification. Within day precisions were 3.0-9.3%, between day precisions 6.0-9.8% and accuracies were between 101 and 110% for the whole calibration range. After validation the assay was used to assess the pharmacokinetics of olaparib in a patient with metastatic breast carcinoma. In addition, systemic exposure of melphalan was monitored in patients subjected to isolated hepatic perfusion with this drug. Both applications show that the new assay can be applied for human pharmacokinetic studies for both drugs. PMID- 21622035 TI - Is the distinction between intra- and extra-musical meaning implemented in the brain? Comment on "Towards a neural basis of processing musical semantics" by Stefan Koelsch. PMID- 21622036 TI - Post-PEG feeding time: a web based national survey amongst gastroenterologists. AB - BACKGROUND: Literature suggests early post-percutaneous endoscopic gastrostomy (PEG) feeding is as safe as delayed feeding. No consensus exists regarding feeding initiation after PEG placement. We performed a national survey to assess current practice regarding feeding initiation after PEG placement as well as clinical and practitioner based factors associated with early or delayed feeding. METHODS: A survey assessing feeding initiation in general ward and intensive care unit patients, current literature knowledge, motility agent use, number of PEG placed per year, along with physician demographics was emailed to 5256 gastroenterologists. Statistical analysis was done using SAS software (V.9). RESULTS: 28% of gastroenterologists responded. Amongst respondents, 59% were private consultants, 25% academic physicians and 16% trainees. Private gastroenterologists initiated feeding earlier (<=12 h) compared to academic gastroenterologists in general ward and intensive care unit patients (p<0.0001). Amongst respondents, 41% of physicians were aware of current literature on post PEG feeding times. Physicians aware of current literature started feeding earlier (<12h) compared to those not aware in general ward and intensive care unit patients (p=0.0002). Male physicians instituted feeding earlier than females (p<0.0001). CONCLUSIONS: Feeding initiation after PEG placement varies amongst gastroenterologists. Further studies are required to explore obstacles in standardizing post-PEG feeding practices. PMID- 21622037 TI - Syphilitic gastritis mimicking gastric neoplasms. PMID- 21622038 TI - A safe and efficient method for intra-operative digital photography using a waterproof case. AB - BACKGROUND: Intra-operative photography provides valuable information for photo documentation. In order to improve quality of photographs and avoid additional contamination, we applied a sterilised waterproof case to adapt a digital camera that allowed the operating surgeon himself to obtain his own ideal images. A prospective study was designed to investigate the efficacy and safety of this technique. MATERIALS AND METHODS: A total of 46 patients were enrolled in this study. The Fujifilm FinePix F30 digital camera encased in Fuji WP-FXF30 waterproof case was used in this study. Microbiological swabs were taken from the case's surface immediately after sealing the digital camera and at the end of surgery. In addition, intra-operative wound swabs were taken for correlation. The patients were followed up to record the possibility of any additional wound infections. RESULTS: None of the swab results on the waterproof case were positive before use. Overall, 11 patients had positive results of bacteria growth from intra-operative wound cultures. Eight of them also revealed positive microorganisms cultured from the case surface after use, in which the bacteria strains were correlated with the intra-operative wound cultures. However, no additional bacteria growth was noted from the culture of case surface. CONCLUSION: A digital camera encased in a sterilised waterproof case met the strict requirements for sterility in our series and demonstrated no added increase in infection rate. Safe use of this technique for obtaining intra operative photographs with high image quality can be achieved. PMID- 21622039 TI - [Feasibility strategy of darbepoetin alfa administration every other week: 2005 2007 experience in a dialysis unit]. AB - Darbepoetin alfa is a recombinant erythropoiesis-stimulating agent, with a longer half-life leaving the possibility to extending dosing administration in haemodialysis patients. A protocol of injection every 2weeks was initiated in the dialysis unit. From 2005 to 2007, 176 dialysis patients were studied with a target haemoglobin level between 11 and 12.5g/dL: the median haemoglobin level was ranged from 11.32 and 11.72g/dL during the study with a median darbepoetin alfa dose injected between 60 and 64MUg per injection (0.46 to 0.47MUg/kg per week). The mean number of dose changes was three per year per patient. The diabetic population did not differ from the general population in terms of haemoglobin levels and doses of darbepoetin alfa. Ten percent of the patients had to resume one injection per week for medical reasons: the profile of these patients was carefully studied. Hospitalisations resulted in a decrease in haemoglobin level and an increase in the darbepoetin alfa doses. Patients who died showed during the last 3months, a particular profile. Hyporesponsiveness has been explored. Time saving with this protocol was important for all the nursing staff. Although numerous factors of variability have been studied, there still a room for improvement of anemia management in haemodialysis patient. PMID- 21622040 TI - Electronic medical records (EMRs), epidemiology, and epistemology: reflections on EMRs and future pediatric clinical research. AB - Electronic medical records (EMRs) are increasingly common in pediatric patient care. EMR data represent a relatively novel and rich resource for clinical research. The fact, however, that pediatric EMR data are collected for the purposes of clinical documentation and billing rather than research creates obstacles to their use in scientific investigation. Particular issues include accuracy, completeness, comparability between settings, ease of extraction, and context of recording. Although these problems can be addressed through standard strategies for dealing with partially accurate and incomplete data, a longer-term solution will involve work with pediatric clinicians to improve data quality. As research becomes one of the explicit purposes for which pediatricians collect EMR data, the pediatric clinician will play a central role in future pediatric clinical research. PMID- 21622041 TI - Specialty referral communication and completion in the community health center setting. AB - OBJECTIVE: Parent and provider disagreement about children's care at the time of specialty referral may lead to incomplete referral, ie, not attending a specialty visit when referred. This study's objectives were first to assess parent-provider correlation in perspectives on referral necessity, seriousness of child's health problem, and parental understanding of referral among children referred to pediatric specialists, and second to assess whether these perspectives are associated with incomplete referral. METHODS: Two months after specialty referral, parents and primary care providers completed a survey rating referral necessity, seriousness of problem, and parental understanding on a 4-part scale ("definitely yes" to "definitely no"). Parents were surveyed by telephone; providers completed one self-administered survey per referral. Using z tests and Pearson correlation coefficients, we summarized parent-provider agreement about referral necessity, seriousness of problem, and parent understanding. We applied logistic regression to test associations of parent and provider ratings for each variable with incomplete referral. RESULTS: A total of 299 (60.0%) of 498 matched parent and provider surveys were included in the analysis. Parents had low correlation with providers in perspectives of referral necessity and seriousness of problem. Parents reported that referral was necessary more often than providers, and providers underestimated parents' self-reported understanding of the referral. Nearly 1 in 3 children had incomplete referral, and both parent and provider reports of lower necessity were associated with incomplete referral. CONCLUSIONS: Parents and providers hold divergent perspectives on referral necessity and seriousness of children's health problems; these perspectives may impact rates of incomplete referral. Improving communication around specialty referral might reduce incomplete referral. PMID- 21622042 TI - White-black disparities in family-centered care among children with autism in the United States: evidence from the NS-CSHCN 2005-2006. AB - OBJECTIVES: The aim of this study was to compare the reported receipt of family centered care between parents of white and black children with autism spectrum disorders (ASD) in the United States, and to disentangle the associations of race and ASD on different aspects of family-centered care. METHODS: Parents of 35,386 children, aged 0 to 17 years, were surveyed by the National Survey of Children with Special Health Care Needs (NS-CSHCN) 2005-2006. Autism was defined by the question, "To the best of your knowledge, does [child] currently have autism or autism spectrum disorder, that is, ASD?" Family-centered care was measured with 5 key indicators on a 4-point Likert scale. Univariate and multivariate analyses were used, with adjustment for the complex sampling design. RESULTS: The prevalence of autism in this sample was 5.4% (n = 1869). We found that, among children with SHCN but no ASD, more white parents than black parents reported receiving family-centered care. Further, fewer parents of both white children and black children with ASD reported receiving family-centered care compared with those with a child who had special needs other than ASD. Lastly, among parents with a child with ASD, being black was associated with lower reporting of family centered care for 3 of 5 items. In multivariate analyses, black parents with a child with ASD had 2 to 5 times greater odds of not reporting family-centered care on each item compared with white parents without a child with ASD. CONCLUSION: Targeted efforts are needed to improve family-centered care for parents with a child with ASD, and particularly for black families. PMID- 21622043 TI - Does information about risks and benefits improve the decision-making process in cancer screening - randomized study. AB - BACKGROUND: Whether the provision of evidence-based information improves satisfaction with decision-making is unclear. OBJECTIVE: To examine whether information about risks and benefits of cancer screening leads to a higher satisfaction with the decision that was made. METHODS: Randomized mail survey in the general population, among 2333 adults aged 30-60 years. The survey included a hypothetical cancer screening scenario that included varying amounts of information about benefits and risks of screening (factorial randomized design). The decision process was evaluated by a 6 item scale, with scores between 0 (lowest score) and 100 (highest score). RESULTS: Substantial proportions of respondents "completely agreed" that the decision reflected what was most important to them (61.2%), were satisfied with their decision (56.0%), were certain of their decision (54.1%), thought that the best choice for them was obvious (53.5%) and that the decision was easy to make (44.1%). The Cronbach alpha coefficient of the scale was 0.88, the mean score was 82.5, and the standard deviation 17.5. Providing information about benefits increased the decision evaluation score only modestly (+1.1, p=0.11); in contrast, providing information about risks sharply reduced the score (-5.1, p<0.001). Those who refused the screening test had lower scores than those who accepted the screening test (69.2 versus 85.6, p<0.001). CONCLUSIONS: Contrary to expectations, informing potential participants about the risks of cancer screening lowered their assessment of the decision process. PMID- 21622044 TI - Expected quality factor of a simple tuned oscillator. AB - A positive feedback system oscillating under self-sustained mode is shown to have an extremely high gain. Modeled as one port, the expected Q is much higher than the loaded Q-factor of the resonator. With just thermal noise present, random phase/frequency deviation is linear. Centered about the oscillator frequency omega/(0), noise frequency on both sides is more amplified with decreasing separation distance. Ultimately, frequency pulling may result in synchronous locking with hysteresis, which occurs because a real oscillator displays a truncated limiting curve. Once locked onto a signal, smaller levels are ignored. A new approach to the design and characterization of a simple tuned oscillator is offered: According to the phenomenon of injection locking, there exists an expected quality factor relating the shape of the truncated limiting curve to an ideal curve. In this paper, synthesis and innovative analytical methods of academic interest are revealed: 1) application of the transducer loss method is revised to establish a new method for oscillator characterization; 2) a transparent method of normalizing a two-port network in the presence of white noise is developed; and 3) in quartz crystal controlled oscillators, characterization of the noise originating from an equivalent noise-resistance determined from parameter of the quartz crystal is proposed. It is shown that the two-port model can also be approximated on a one-port basis. In conclusion, a sample of closed-form estimation of expected Q-factor order of magnitude of piezoelectric resonator oscillators is calculated. PMID- 21622045 TI - Advanced noise reduction techniques for ultra-low phase noise optical-to microwave division with femtosecond fiber combs. AB - We report what we believe to be the lowest phase noise optical-to-microwave frequency division using fiber-based femtosecond optical frequency combs: a residual phase noise of -120 dBc/Hz at 1 Hz offset from an 11.55 GHz carrier frequency. Furthermore, we report a detailed investigation into the fundamental noise sources which affect the division process itself. Two frequency combs with quasi-identical configurations are referenced to a common ultrastable cavity laser source. To identify each of the limiting effects, we implement an ultra-low noise carrier-suppression measurement system, which avoids the detection and amplification noise of more conventional techniques. This technique suppresses these unwanted sources of noise to very low levels. In the Fourier frequency range of ~200 Hz to 100 kHz, a feed-forward technique based on a voltage controlled phase shifter delivers a further noise reduction of 10 dB. For lower Fourier frequencies, optical power stabilization is implemented to reduce the relative intensity noise which causes unwanted phase noise through power-to-phase conversion in the detector. We implement and compare two possible control schemes based on an acousto-optical modulator and comb pump current. We also present wideband measurements of the relative intensity noise of the fiber comb. PMID- 21622046 TI - Magnetic tracking of acoustic radiation force-induced micro-order displacement. AB - The dynamic behavior of a rigid magnetic sphere induced by an acoustic radiation force was investigated. The sphere was suspended in water in a simple pendulum configuration. The drag force acting on the pendulum during its motion was considered to follow a modified Stokes law for a low Reynolds number, accounting for phenomena related to its oscillatory movement. Steady forces of long (a few seconds) and short (a few milliseconds) durations were used. The movement of the magnetic sphere was tracked using a magnetoresistive sensor. From the new equilibrium position of the sphere in response to the long-duration static radiation force, the amplitude of this force was estimated. To assess the water viscosity, the relaxation movement after the acoustic force had stopped was fitted to a harmonic-motion model. Based on the results for the acoustic force and water viscosity, a theoretical profile of the sphere's micro-order displacement as a function of time caused by short-duration acoustic radiation force agreed well with experimental results. PMID- 21622047 TI - 1.5-GHz voltage controlled oscillator with 3% tuning bandwidth using a two-pole DSBAR filter. AB - First results on a novel voltage controlled oscillator (VCO) in the lower gigahertz range, featuring excellent phase noise and high power efficiency are presented. The heart of the VCO is a recently reported novel miniature two-pole decoupled stacked bulk acoustic resonator (DSBAR) filter. With its single 180 degrees phase transition over the 1 dB bandwidth, linear phase, and maximum 1 dB insertion loss, it provides stable single-mode operation over 45 MHz (~3%) of tuning bandwidth and has negligible heat dissipation when operated at incident power levels of 100 mW or greater. The 1.55-GHz laboratory VCO prototypes operate at 5 V supply voltage, 50 mA supply current, 15 dBm of output power, and >13% efficiency, demonstrating -84 and < -180 dBc/Hz phase noise suppression at 1 kHz carrier offset and in the thermal noise region, respectively. VCOs with cascaded DSBAR filters for further phase noise reduction are also demonstrated. PMID- 21622048 TI - Parametric array technique for microbubble excitation. AB - This study investigates the use of an acoustic parametric array as a means for microbubble excitation. The excitation wave is generated during propagation in a nonlinear medium of two high-frequency carrier waves, whereby the frequency of the excitation wave is the difference frequency of the carrier waves. Carrier waves of around 10 and 25 MHz are used to generate low-frequency waves between 0.5 and 3.5 MHz at amplitudes in the range of 25 to 80 kPa in water. We demonstrate with high-speed camera observations that it is possible to induce microbubble oscillations with the low frequency signal arising from the nonlinear propagation process. As an application, we determined the resonance frequency of Definity contrast agent microbubbles with radius ranging from 1.5 to 5 MUm by sweeping the difference frequency in the range from 0.5 to 3.5 MHz. PMID- 21622049 TI - Synthetic aperture focusing for a single-element transducer undergoing helical motion. AB - This paper describes the application of 3-D synthetic aperture focusing (SAF) to a single-element trans-rectal ultrasound transducer. The transducer samples a 3-D volume by simultaneous rotation and translation, giving a helical motion. Two different 3-D SAF methods are investigated, a direct and a two-step approach. Both methods perform almost identically for simulated scatterers and give a significant improvement in azimuth resolution and a constant resolution in elevation. Side lobes below -60 dB are achievable for both methods. Validation of the method is achieved by scanning a simple wire phantom and a complex phantom containing wires in azimuth and elevation. The simple wire phantom shows the same results as that found through simulation. The complex phantom shows simultaneous focusing in azimuth and elevation for the wire scatterers. Consideration of the processing requirements for both 3-D SAF methods shows that the two-step approach can give equivalent performance using an order of magnitude fewer calculations. This reduction requires a temporary storage of 9.1 GB of data for the investigated setup. PMID- 21622050 TI - Ultrasound therapy transducers with space-filling non-periodic arrays. AB - Ultrasound transducers designed for therapeutic purposes such as tissue ablation, histotripsy, or drug delivery require large apertures for adequate spatial localization while providing sufficient power and steerability without the presence of secondary grating lobes. In addition, it is highly preferred to minimize the total number of channels and to maintain simplicity in electrical matching network design. To this end, we propose array designs that are both space-filling and non-periodic in the placement of the elements. Such array designs can be generated using the mathematical concept of non-periodic or aperiodic tiling (tessellation) and can lead to reduced grating lobes while maintaining full surface area coverage to deliver maximum power. For illustration, we designed two 2-D space-filling therapeutic arrays with 128 elements arranged on a spherical shell. One was based on the two-shape Penrose rhombus tiling, and the other was based on a single rectangular shape arranged non-periodically. The steerability performance of these arrays was studied using acoustic field simulations. For comparison, we also studied two other arrays, one with circular elements distributed randomly, and the other a periodic array with square elements. Results showed that the two space-filling non-periodic arrays were able to steer to treat a volume of 16 x 16 x 20 mm while ensuring that the grating lobes were under -10 dB compared with the main lobe. The rectangular non periodic array was able to generate two and half times higher power than the random circles array. The rectangular array was then fabricated by patterning the array using laser scribing methods and its steerability performance was validated using hydrophone measurements. This work demonstrates that the concept of space filling aperiodic/non-periodic tiling can be used to generate therapy arrays that are able to provide higher power for the same total transducer area compared with random arrays while maintaining acceptable grating lobe levels. PMID- 21622051 TI - Microbubble sizing and shell characterization using flow cytometry. AB - Experiments were performed to size, count, and obtain shell parameters for individual ultrasound contrast microbubbles using a modified flow cytometer. Light scattering was modeled using Mie theory, and applied to calibration beads to calibrate the system. The size distribution and population were measured directly from the flow cytometer. The shell parameters (shear modulus and shear viscosity) were quantified at different acoustic pressures (from 95 to 333 kPa) by fitting microbubble response data to a bubble dynamics model. The size distribution of the contrast agent microbubbles is consistent with manufacturer specifications. The shell shear viscosity increases with increasing equilibrium microbubble size, and decreases with increasing shear rate. The observed trends are independent of driving pressure amplitude. The shell elasticity does not vary with microbubble size. The results suggest that a modified flow cytometer can be an effective tool to characterize the physical properties of microbubbles, including size distribution, population, and shell parameters. PMID- 21622052 TI - Pulsed high-intensity focused ultrasound enhances the relative permeability of the blood-tumor barrier in a glioma-bearing rat model. AB - The use of pulsed high-intensity focused ultrasound (HIFU) with an ultrasound contrast agent (UCA) has been shown to disrupt the blood-brain barrier (BBB) noninvasively and reversibly in the targeted regions. This study evaluated the relative permeability of the blood-tumor barrier (BTB) after sonication by pulsed HIFU. Entry into the brain of chemotherapeutic agents is impeded by the BBB even though the permeability of this barrier may be partially reduced in the presence of a brain tumor. F98 glioma-bearing rats were injected intravenously with Evans blue (EB) with or without BTB disruption induced by pulsed HIFU. Sonication was applied at an ultrasound frequency of 1 MHz with a 5% duty cycle, and a repetition frequency of 1 Hz. The accumulation of EB in brain tumor and the tumor to-contralateral brain ratio of EB were highest after pulsed HIFU exposure. Sonication followed by EB injection showed a tumor-to-contralateral brain ratio in the target tumors which was about 2 times that of the control tumors. This research demonstrates that pulsed HIFU enhances the relative permeability of the BTB in glioma- bearing rats. The results of this pilot study support the idea that further evaluation of other treatment strategies, such as HIFU exposure in addition to combined chemotherapy or repeated pulsed HIFU exposure to increase delivery of drugs into brain tumors, might be useful. PMID- 21622053 TI - In vitro comminution of model renal calculi using histotripsy. AB - Shock wave lithotripsy (SWL) suffers from the fact that it can produce residual stone fragments of significant size (>2 mm). Mechanistically, cavitation has been shown to play an important role in the reduction of such fragments to smaller debris. In this study, we assessed the feasibility of using cavitationally-based pulsed ultrasound therapy (histotripsy) to erode kidney stones. Previous work has shown that histotripsy is capable of mechanically fractionating soft tissue into fine, acellular debris. Here, we investigated the potential for translating this technology to renal calculi through the use of a commonly accepted stone model. Stone models were sonicated using a 1-MHz focused transducer, with 5-cycle pulses delivered at a rate of 1 kHz. Pulses having peak negative pressures ranging from 3 to 21 MPa were tested. Results indicate that histotripsy is capable of effectively eroding the stone model, achieving an average stone erosion rate of 26 mg/min at maximum treatment pressure; substantial stone erosion was only observed in the presence of a dense cavitational bubble cloud. Sequential sieving of residual stone fragments indicated that debris produced by histotripsy was smaller than 100 MUm in size, and treatment monitoring showed that both the cavitational bubble cloud and model stone appear as hyperechoic regions on B-mode imaging. These preliminary results indicate that histotripsy shows promise in its use for stone comminution, and an optimized erosion process may provide a potential adjunct to conventional SWL procedures. PMID- 21622054 TI - Review of shell models for contrast agent microbubbles. AB - Micrometer-scale encapsulated gas bubbles, known as ultrasound contrast agents, are used in ultrasound medical diagnostics for enhancing blood-tissue contrast during an ultrasonic examination. They are also employed in therapy as an activator of drug incorporation or extravasation. Adequate modeling of the effect of encapsulation is of primary importance because it is the encapsulating shell that determines many of the functional properties of contrast agents. In this review, existing approaches to the modeling of the radial motion of an encapsulated bubble are discussed and comparative analysis of available shell models is conducted. The capabilities of the shell models are evaluated in the context of recent experimental observations, such as compression-only behavior and the dependence of shell material properties on initial bubble radius. It is shown that for early shell models, the main problem is that the behavior of encapsulation is described by linear elastic and viscous laws, whereas recent experimental data attest to complicated rheological properties inherent in shell materials. Currently, a trend toward models involving nonlinear laws for shell elasticity and viscosity is observed. In particular, nonlinear models have been proposed that allow one to reproduce compression-only behavior. However, the problem of the radius dependence of shell material parameters remains unsolved. PMID- 21622056 TI - Phase coherence imaging of grained materials. AB - Ultrasound detection and evaluation of flaws in materials showing structural noise (austenitic steels, titanium alloys, composites, etc.) is difficult because of the low flaw-to-grain noise ratio. Much research has been performed looking for methods to improve flaw detection in grained materials. Many approaches require a cumbersome tuning process to select the correct parameter values or to use iterative techniques. In this work, the technique of phase coherence imaging is proposed to improve the flaw-to-grain noise ratio. The technique weights the output of a conventional beamformer with a coherence factor obtained from the aperture data phase dispersion. It can be simply implemented in real-time and it operates automatically, without needing any parameter adjustment. This paper presents the theoretical basis of phase coherence imaging to reduce grain noise, as well as experimental results that confirm the expected performance. PMID- 21622055 TI - Characterization of the spatial resolution of different high-frequency imaging systems using a novel anechoic-sphere phantom. AB - The spatial resolution of high-frequency ultrasound (HFU, >20 MHz) imaging systems is usually determined using wires perpendicular to the beam. Recently, two tissue-mimicking phantoms (TMPs) were developed to estimate three-dimensional (3-D) resolution. Each TMP consists of nine 1-cm-wide slabs of tissue-mimicking material containing randomly distributed anechoic spheres. All anechoic spheres in one slab have the same dimensions, and their diameter is increased from 0.1 mm in the first slab to 1.09 mm in the last. The scattering background for one set of slabs was fabricated using 3.5-MUm glass beads; the second set used 6.4-MUm glass beads. The ability of a HFU system to detect these spheres against a speckle background provides a realistic estimation of its 3-D spatial resolution. In the present study, these TMPs were used with HFU systems using single-element transducers, linear arrays, and annular arrays. The TMPs were immersed in water and each slab was scanned using two commercial imaging systems and a custom HFU system based on a 5-element annular array. The annular array had a nominal center frequency of 40 MHz, a focal length of 12 mm, and a total aperture of 6 mm. A synthetic-focusing algorithm was used to form images with an increased depth-of field. The penetration depth was increased by using a linear-chirp signal spanning 15 to 65 MHz over 4 MUs. Results obtained with the custom system were compared with those of the commercial systems (40-MHz probes) in terms of sphere detection, i.e., 3-D spatial resolution, and contrast-to-noise ratio (CNR). Resulting B-mode images indicated that only the linear-array transducer failed to clearly resolve the 0.2-mm spheres, which showed that the 3-D spatial resolution of the single-element and annular-array transducers was superior to that of the linear array. The single-element transducer could only detect these spheres over a narrow 1.5 mm depth-of-field, whereas the annular array was able to detect them to depths of at least 7 mm. For any size of the anechoic spheres, the annular array excited by a chirp-coded signal provided images of the highest contrast, with a maximum CNR of 1.8 at the focus, compared with 1.3 when using impulse excitation and 1.6 with the single-element transducer and linear array. This imaging configuration also provided CNRs above 1.2 over a wide depth range of 8 mm, whereas CNRs would quickly drop below 1 outside the focal zone of the other configurations. PMID- 21622057 TI - An analytical model for guided wave inspection optimization for prismatic structures of any cross section. AB - This paper presents an analytical modeling technique for the simulation of long range ultrasonic guided waves in structures. The model may be used to predict the displacement field in a prismatic structure arising from any excitation arrangement and may therefore be used as a tool to design new inspection systems. It is computationally efficient and relatively simple to implement, yet gives accuracy similar to finite element analysis and semi-analytical finite element analysis methods. The model has many potential applications; one example is the optimization of part-circumferential arrays where access to the full circumference of the pipe is restricted. The model has been successfully validated by comparison with finite element solutions. Experimental validation has also been carried out using an array of piezoelectric transducer elements to measure the displacement field arising from a single transducer element in an 88.9-mm-diameter pipe. Good agreement has been obtained between the two models and the experimental data. PMID- 21622058 TI - EMAT noise suppression using information fusion in stationary wavelet packets. AB - An important issue in ultrasonic nondestructive testing is the detection of flaw echoes in the presence of background noise created by instrumentation and by clutter noise. Signal averaging, autoregressive analysis, spectrum analysis, matched filtering, and the wavelet transform have all been used to filter noise in ultrasonic signals. Widely-used wavelet threshold estimation algorithms are not designed for electromagnetic acoustic transducer (EMAT) pulse-echo signals, and therefore do not exploit their unique impulse nature. The approach to ultrasonic signal filtering proposed in this paper is based on stationary wavelet packet denoising with a threshold influenced by several information sources: a statistical echo detection, the amplitude distribution of the wavelet transform coefficients, and a priori known system frequency characteristics. The proposed method was evaluated on signals measured with EMAT probes and under various SNR conditions; it outperforms the wavelet transform with the Stein unbiased risk estimate (SURE) threshold estimation method and split-spectrum processing (SSP). The results indicate SNR enhancement of 19 dB with real EMAT data. PMID- 21622059 TI - Synthetic aperture focusing of ultrasonic data from multilayered media using an omega-k algorithm. AB - The synthetic aperture focusing technique (SAFT) is used to create focused images from ultrasound scans. SAFT has traditionally been applied only for imaging in a single medium, but the recently introduced phase shift migration (PSM) algorithm has expanded the use of SAFT to multilayer structures. In this article we present a similar focusing algorithm called multi-layer omega-k (MULOK), which combines PSM and the omega-k algorithm to perform multilayer imaging more efficiently. The asymptotic complexity is shown to be lower for MULOK than for PSM, and this is confirmed by comparing execution times for implementations of both algorithms. To facilitate the complexity analysis, a detailed description of algorithm implementation is included, which also serves as a guide for readers interested in practical implementation. Using data from an experiment with a multilayered structure, we show that there is essentially no difference in image quality between the two algorithms. PMID- 21622060 TI - A single vibration mode tubular piezoelectric ultrasonic motor. AB - A novel tubular ultrasonic motor is presented that uses only a single vibration bending mode of a piezoelectric tube to generate rotation. When the piezoelectric tube bends, the diagonal motion of points on selected areas at the ends of the tube generates forces with tangential components along the same circumferential direction, driving the rotors to rotate. Bi-directional motion is achieved by simply switching the direction of bending. Because only one vibration mode is used, the motor requires only one driving signal and no vibration mode coupling is needed, simplifying the design, fabrication, assembly, and operation of the device. Two prototypes [one with cut-in lead zirconate titanate (PZT) teeth and one with added metal teeth] were built and tested using PZT tubes available to the authors. The tubes have an outside diameter of 6.6 mm, inner diameter of 5.0 mm, and length of 25.4 mm. The working frequencies of the two motors are 27.6 and 23.5 kHz. The motors achieved a maximum no-load speed of 400 rpm and a stall torque of 300 MUN.m. PMID- 21622061 TI - c-Axis zig-zag ZnO film ultrasonic transducers for designing longitudinal and shear wave resonant frequencies and modes. AB - A method for designing frequencies and modes in ultrasonic transducers above the very-high-frequency (VHF) range is required for ultrasonic non-destructive evaluation and acoustic mass sensors. To obtain the desired longitudinal and shear wave conversion loss characteristics in the transducer, we propose the use of a c-axis zig-zag structure consisting of multilayered c-axis 23 degrees tilted ZnO piezoelectric films. In this structure, every layer has the same thickness, and the c-axis tilt directions in odd and even layers are symmetric with respect to the film surface normal. c-axis zig-zag crystal growth was achieved by using a SiO(2) low-temperature buffer layer. The frequency characteristics of the multilayered transducer were predicted using a transmission line model based on Mason's equivalent circuit. We experimentally demonstrated two types of transducers: those exciting longitudinal and shear waves simultaneously at the same frequency, and those exciting shear waves with suppressed longitudinal waves. PMID- 21622062 TI - An improved equivalent circuit model of radial mode piezoelectric transformer. AB - In this paper, both the equivalent circuit models of the radial mode and the coupled thickness vibration mode of the radial mode piezoelectric transformer are deduced, and then with the Y-parameter matrix method and the dual-port network theory, an improved equivalent circuit model for the multilayer radial mode piezoelectric transformer is established. A radial mode transformer sample is tested to verify the equivalent circuit model. The experimental results show that the model proposed in this paper is more precise than the typical model. PMID- 21622063 TI - Perfectly matched layers for frequency-domain integral equation acoustic scattering problems. AB - Simulations of acoustic wavefields in inhomogeneous media are always performed on finite numerical domains. If contrasts actually extend over the domain boundaries of the numerical volume, unwanted, non-physical reflections from the boundaries will occur. One technique to suppress these reflections is to attenuate them in a locally reflectionless absorbing boundary layer enclosing the spatial computational domain, a perfectly matched layer (PML). This technique is commonly applied in time-domain simulation methods like finite element methods or finite difference time-domain, but has not been applied to the integral equation method. In this paper, a PML formulation for the three-dimensional frequency-domain integral-equation-based acoustic scattering problem is derived. Three-dimensional acoustic scattering configurations are used to test the PML formulation. The results demonstrate that strong attenuation (a factor of 200 in amplitude) of the scattered pressure field is achieved for thin layers with a thickness of less than a wavelength, and that the PMLs themselves are virtually reflectionless. In addition, it is shown that the integral equation method, both with and without PMLs, accurately reproduces pressure fields by comparing the obtained results with analytical solutions. PMID- 21622064 TI - Improved SHSAW transduction efficiency using grating and uniform electrode guiding. AB - Pure shear-horizontal surface acoustic wave (SHSAW) devices have been increasingly considered for liquidphase and biosensing applications because of their ability to operate under liquid-loaded conditions and intrinsic sensitivity to mass, stiffness, viscosity, and electrical perturbations occurring at the device/fluid interface. Typically, the SHSAW is weakly guided by a free surface boundary condition (BC) or may not even exist for some materials and orientations, such as the quartz ST-90 degrees orientation considered in this work. For a surrounding free surface BC, the interdigital transducer (IDT) typically generates strong shear-horizontal bulk acoustic waves (SHBAWs) relative to SHSAW. For that reason, guiding structures, e.g., dense and/or thick electrodes in periodic or uniform configurations, are incorporated into the design and placed between IDTs in delay-line devices to increase the ratio of transduced SHSAW power to IDT input power, etaSHSAW. The degree of etaSHSAW improvement depends on the thickness, composition, and geometry of the guiding structure. In previous work, the authors evaluated etaSHSAW using hybrid finite and boundary element method (FEM/BEM) models, but were limited to cases of stress free or finite-thickness-grating surrounding surfaces. This work extends the analysis to the important boundary condition case of uniform finite-thickness electrode guiding, which is typically employed in liquid-phase and biosensor applications. To integrate the uniform electrode guiding structure with the SHSAW device analysis, a combined finite-length uniform electrode structure followed by an additional quarter-wavelength electrode was considered. In this work, it is shown that adjusting the quarter-wavelength electrode's film thickness and length allows cancellation of the SHSAW reflection from the edge discontinuity. As a result, the finite-length uniform guiding electrode can be treated as if it extends to infinity, and etaSHSAW can be easily obtained. In addition, the finite thickness of all electrodes is considered in the calculations. To verify the model, an IDT with uniform guiding electrodes was simulated and compared with the experimental results of a fabricated and tested device. The simulations predict SHSAW excitation directivity of 9 dB by the IDT, which is experimentally confirmed to within 0.8 dB. PMID- 21622065 TI - Verification of the Westervelt equation for focused transducers. AB - This study investigates the validity of the Westervelt equation for focused transducers. The angular spectrum method is employed to analyze the second harmonic acoustic field under the weakly nonlinear approximation. Although it is well known that the Westervelt equation is accurate for the case of quasi-plane waves, the present work demonstrates accurate solution for the highly focused case of a spherically-curved ultrasound transducer having an aperture angle of 80 degrees . It is further found that the solution error is inversely dependent on the nonlinearity coefficient. PMID- 21622066 TI - Resonant frequency function of thickness-shear vibrations of rectangular crystal plates. AB - The resonant frequencies of thickness-shear vibrations of quartz crystal plates in rectangular and circular shapes are always required in the design and manufacturing of quartz crystal resonators. As the size of quartz crystal resonators shrinks, for rectangular plates we must consider effects of both length and width for the precise calculation of resonant frequency. Starting from the three-dimensional equations of wave propagation in finite crystal plates and the general expression of vibration modes, we obtained the relations between frequency and wavenumbers. By satisfying the major boundary conditions of the dominant thickness-shear mode, three wavenumber solutions are obtained and the frequency equation is constructed. It is shown the resonant frequency of thickness-shear mode is a second-order polynomial of aspect ratios. This conforms to known results in the simplest form and is applicable to further analytical and experimental studies of the frequency equation of quartz crystal resonators. PMID- 21622067 TI - Deconvolved spatial light interference microscopy for live cell imaging. AB - Spatial light interference microscopy (SLIM) is a recently developed method for the label-free imaging of live cells, using the quantitative optical path length through the sample as an endogenous source of contrast. In conventional SLIM, spatial resolution is limited by diffraction and aberrations. This paper describes a novel constrained deconvolution method for improving resolution in SLIM. Constrained deconvolution is enabled by experimental measurement of the system point-spread function and the modeling of coherent image formation in SLIM. Results using simulated and experimental data demonstrate that the proposed method leads to significant improvements in the resolution and contrast of SLIM images. The proposed method should prove useful for high-resolution label-free studies of biological cells and subcellular processes. PMID- 21622068 TI - Dielectric characterization of PCL-based thermoplastic materials for microwave diagnostic and therapeutic applications. AB - We propose the use of a polycaprolactone (PCL)-based thermoplastic mesh as a tissue-immobilization interface for microwave imaging and microwave hyperthermia treatment. An investigation of the dielectric properties of two PCL-based thermoplastic materials in the frequency range of 0.5-3.5 GHz is presented. The frequency-dependent dielectric constant and effective conductivity of the PCL based thermoplastics are characterized using measurements of microstrip transmission lines fabricated on substrates comprised of the thermoplastic meshes. We also examine the impact of the presence of a PCL-based thermoplastic mesh on microwave breast imaging. We use a numerical test bed comprised of a previously reported 3-D anatomically realistic breast phantom and a multi frequency microwave inverse scattering algorithm. We demonstrate that the PCL based thermoplastic material and the assumed biocompatible medium of vegetable oil are sufficiently well matched such that the PCL layer may be neglected by the imaging solution without sacrificing imaging quality. Our results suggest that PCL-based thermoplastics are promising materials as tissue immobilization structures for microwave diagnostic and therapeutic applications. PMID- 21622069 TI - An automatic spike detection system based on elimination of false positives using the large-area context in the scalp EEG. AB - Most automatic spike detection systems in the scalp electroencephalogram (EEG) focused on the characteristics of "spike." However, the characteristics of "false positives" (FPs) have not been fully studied. In this paper, we proposed a system that contains a series of algorithms to eliminate FPs and a template method to confirm spikes. The system used large area context available on 49 channels from two common montages. The impact of slow-waves after spikes was taken into consideration, as well as the information from single channel, multichannel, and whole recording. Two types of FPs were identified in this paper. The ones from typical artifacts were identified by analysis of background EEG activities, and the ones from other EEG activities were declared by spatial and temporal characteristics of spike activities. Finally, a multichannel template method was used to assess the performance of the proposed system. The system was evaluated using 17 routine EEG recordings. Spike activities were observed in six of them. Effective multichannel templates were extracted from four recordings containing frequent spikes. The least selectivity was 92.6% and the most false positive rate was 0.26 min(-1). Proposed algorithms for elimination of FPs are also suitable for other algorithms to enhance performance since most FPs can be identified while few true spikes are eliminated. PMID- 21622070 TI - Respiration effect on wavelet-based ECG T-wave end delineation strategies. AB - The main purpose of this paper is to study the influence of the mechanical effect of respiration over the T-wave end delineation. We compared the performance of an automatic delineation system based on the wavelet transform (WT), considering single lead (SL), global delineation locations obtained from SL annotations (SLR), and multilead (ML) approaches. The linear relation between the variations on T-wave end locations obtained with each of the methods and the mechanical effect of respiration was quantified using spectral coherence and ARARX modeling both in simulated signals and in real data. We also explored the evolution of the vectorcardiographic spatial loop using the projection on the main direction of the WT in the region close to the T-wave end ( T(e)) and its relation with respiration. The dispersion of the additional T-wave end location error due to respiration was reduced by 15% using SLR with respect to SL, while ML allows for a reduction of around 40%. The percentage of that error correlated with respiration was in average 99% using SL while 82% and 72% using SLR and ML, respectively. Thus, results suggest that ML is the most adequate strategy for T wave delineation, allowing the reduction of the instability of T-wave end location caused by respiration. PMID- 21622071 TI - An insulin infusion advisory system based on autotuning nonlinear model predictive control. AB - This paper aims at the development and evaluation of a personalized insulin infusion advisory system (IIAS), able to provide real-time estimations of the appropriate insulin infusion rate for type 1 diabetes mellitus (T1DM) patients using continuous glucose monitors and insulin pumps. The system is based on a nonlinear model-predictive controller (NMPC) that uses a personalized glucose insulin metabolism model, consisting of two compartmental models and a recurrent neural network. The model takes as input patient's information regarding meal intake, glucose measurements, and insulin infusion rates, and provides glucose predictions. The predictions are fed to the NMPC, in order for the latter to estimate the optimum insulin infusion rates. An algorithm based on fuzzy logic has been developed for the on-line adaptation of the NMPC control parameters. The IIAS has been in silico evaluated using an appropriate simulation environment (UVa T1DM simulator). The IIAS was able to handle various meal profiles, fasting conditions, interpatient variability, intraday variation in physiological parameters, and errors in meal amount estimations. PMID- 21622072 TI - Automatic aneurysm neck detection using surface Voronoi diagrams. AB - A new automatic approach for saccular intracranial aneurysm isolation is proposed in this work. Due to the inter- and intra-observer variability in manual delineation of the aneurysm neck, a definition based on a minimum cost path around the aneurysm sac is proposed that copes with this variability and is able to make consistent measurements along different data sets, as well as to automate and speedup the analysis of cerebral aneurysms. The method is based on the computation of a minimal path along a scalar field obtained on the vessel surface, to find the aneurysm neck in a robust and fast manner. The computation of the scalar field on the surface is obtained using a fast marching approach with a speed function based on the exponential of the distance from the centerline bifurcation between the aneurysm dome and the parent vessels. In order to assure a correct topology of the aneurysm sac, the neck computation is constrained to a region defined by a surface Voronoi diagram obtained from the branches of the vessel centerline. We validate this method comparing our results in 26 real cases with manual aneurysm isolation obtained using a cut-plane, and also with results obtained using manual delineations from three different observers by comparing typical morphological measures. PMID- 21622073 TI - Analysis and compensation of the effects of analog VLSI arithmetic on the LMS algorithm. AB - Analog very large scale integration implementations of neural networks can compute using a fraction of the size and power required by their digital counterparts. However, intrinsic limitations of analog hardware, such as device mismatch, charge leakage, and noise, reduce the accuracy of analog arithmetic circuits, degrading the performance of large-scale adaptive systems. In this paper, we present a detailed mathematical analysis that relates different parameters of the hardware limitations to specific effects on the convergence properties of linear perceptrons trained with the least-mean-square (LMS) algorithm. Using this analysis, we derive design guidelines and introduce simple on-chip calibration techniques to improve the accuracy of analog neural networks with a small cost in die area and power dissipation. We validate our analysis by evaluating the performance of a mixed-signal complementary metal-oxide semiconductor implementation of a 32-input perceptron trained with LMS. PMID- 21622074 TI - Manifold Learning by Graduated Optimization. AB - We present an algorithm for manifold learning called manifold sculpting , which utilizes graduated optimization to seek an accurate manifold embedding. An empirical analysis across a wide range of manifold problems indicates that manifold sculpting yields more accurate results than a number of existing algorithms, including Isomap, locally linear embedding (LLE), Hessian LLE (HLLE), and landmark maximum variance unfolding (L-MVU), and is significantly more efficient than HLLE and L-MVU. Manifold sculpting also has the ability to benefit from prior knowledge about expected results. PMID- 21622075 TI - Gait-based gender classification using mixed conditional random field. AB - This paper proposes a supervised modeling approach for gait-based gender classification. Different from traditional temporal modeling methods, male and female gait traits are competitively learned by the addition of gender labels. Shape appearance and temporal dynamics of both genders are integrated into a sequential model called mixed conditional random field (CRF) (MCRF), which provides an open framework applicable to various spatiotemporal features. In this paper, for the spatial part, pyramids of fitting coefficients are used to generate the gait shape descriptors; for the temporal part, neighborhood preserving embeddings are clustered to allocate the stance indexes over gait cycles. During these processes, we employ evaluation functions like the partition index and Xie and Beni's index to improve the feature sparseness. By fusion of shape descriptors and stance indexes, the MCRF is constructed in coordination with intra- and intergender temporary Markov properties. Analogous to the maximum likelihood decision used in hidden Markov models (HMMs), several classification strategies on the MCRF are discussed. We use CASIA (Data set B) and IRIP Gait Databases for the experiments. The results show the superior performance of the MCRF over HMMs and separately trained CRFs. PMID- 21622076 TI - Accurate landmarking of three-dimensional facial data in the presence of facial expressions and occlusions using a three-dimensional statistical facial feature model. AB - Three-dimensional face landmarking aims at automatically localizing facial landmarks and has a wide range of applications (e.g., face recognition, face tracking, and facial expression analysis). Existing methods assume neutral facial expressions and unoccluded faces. In this paper, we propose a general learning based framework for reliable landmark localization on 3-D facial data under challenging conditions (i.e., facial expressions and occlusions). Our approach relies on a statistical model, called 3-D statistical facial feature model, which learns both the global variations in configurational relationships between landmarks and the local variations of texture and geometry around each landmark. Based on this model, we further propose an occlusion classifier and a fitting algorithm. Results from experiments on three publicly available 3-D face databases (FRGC, BU-3-DFE, and Bosphorus) demonstrate the effectiveness of our approach, in terms of landmarking accuracy and robustness, in the presence of expressions and occlusions. PMID- 21622077 TI - A novel emotion elicitation index using frontal brain asymmetry for enhanced EEG based emotion recognition. AB - This paper aims at providing a novel method for evaluating the emotion elicitation procedures in an electroencephalogram (EEG)-based emotion recognition setup. By employing the frontal brain asymmetry theory, an index, namely asymmetry Index (AsI), is introduced, in order to evaluate this asymmetry. This is accomplished by a multidimensional directed information analysis between different EEG sites from the two opposite brain hemispheres. The proposed approach was applied to three-channel (Fp1, Fp2, and F3/F4 10/20 sites) EEG recordings drawn from 16 healthy right-handed subjects. For the evaluation of the efficiency of the AsI, an extensive classification process was conducted using two feature-vector extraction techniques and a SVM classifier for six different classification scenarios in the valence/arousal space. This resulted in classification results up to 62.58% for the user independent case and 94.40% for the user-dependent one, confirming the efficacy of AsI as an index for the emotion elicitation evaluation. PMID- 21622078 TI - Automatic skin lesion segmentation via iterative stochastic region merging. AB - An automatic method for segmenting skin lesions in conventional macroscopic images is presented. The images are acquired with conventional cameras, without the use of a dermoscope. Automatic segmentation of skin lesions from macroscopic images is a very challenging problem due to factors such as illumination variations, irregular structural and color variations, the presence of hair, as well as the occurrence of multiple unhealthy skin regions. To address these factors, a novel iterative stochastic region-merging approach is employed to segment the regions corresponding to skin lesions from the macroscopic images, where stochastic region merging is initialized first on a pixel level, and subsequently on a region level until convergence. A region merging likelihood function based on the regional statistics is introduced to determine the merger of regions in a stochastic manner. Experimental results show that the proposed system achieves overall segmentation error of under 10% for skin lesions in macroscopic images, which is lower than that achieved by existing methods. PMID- 21622079 TI - Hand Spring Operated Movement Enhancer (HandSOME): a portable, passive hand exoskeleton for stroke rehabilitation. AB - Stroke patients often have flexor hypertonia and finger extensor weakness, which makes it difficult to open their affected hand for functional grasp. Because of this impairment, hand rehabilitation after stroke is essential for restoring functional independent lifestyles. The goal of this study is to develop a passive, lightweight, wearable device to assist with hand function during performance of activities of daily living. The device, Hand Spring Operated Movement Enhancer (HandSOME), assists with opening the patient's hand using a series of elastic cords that apply extension torques to the finger joints and compensates for the flexor hypertonia. Device design and calibration are described as well as functional and usability testing with stroke subjects with a wide range of hand impairments. In initial testing with eight stroke subjects with finger flexor hypertonia, use of the HandSOME significantly increased range of motion and functional ability (p=0.002) . There was some decrease in grip strength with the HandSOME device at the subject's ideal setting, however this was not statistically significant (p=0.167) and did not seem to have a significant effect on function. Overall HandSOME shows promise as a training tool to facilitate repetitive task practice for improving hand function in stroke patients. HandSOME can be used as part of a home-based therapy program, or as an orthotic for replacing lost function. PMID- 21622080 TI - Automatic seizure detection in ECoG by differential operator and windowed variance. AB - Differential operator has long been used in image and signal processing with great success to detect significant changes. In this paper we show that differentiation can enhance certain features of brain electrophysiological signals, contaminated with noise, artifacts, and acquisition defects, leading to efficient detection of those changes. Windowed variance method has been very successful in detecting seizure onset in the brain electrophysiological signals. In this paper we have combined these two powerful methods under the name of differential windowed variance (DWV) algorithm to automatically detect seizure onsets in almost real time, in continuous ECoG (depth-EEG) signals of epileptic patients. The main advantages of the method are simplicity of implementation and speed. We have tested the algorithm on 369 h of nonseizure ECoG as well as 59 h of seizure ECoG of 15 epileptic patients. It detected all but six seizures (91.525% accuracy) with an average delay of 9.2 s after the onset with a maximum false detection of three in 24 h of nonseizure data. Eight novel empirical measures have been introduced to avoid false detections. To ascertain the reliability of the detection method a novel methodology, called quasi-ROC (qROC) curve analysis has been introduced. DWV has been compared with a difference filter based sharp transient (ST) detection algorithm. PMID- 21622081 TI - Revealing ensemble state transition patterns in multi-electrode neuronal recordings using hidden Markov models. AB - In order to harness the computational capacity of dissociated cultured neuronal networks, it is necessary to understand neuronal dynamics and connectivity on a mesoscopic scale. To this end, this paper uncovers dynamic spatiotemporal patterns emerging from electrically stimulated neuronal cultures using hidden Markov models (HMMs) to characterize multi-channel spike trains as a progression of patterns of underlying states of neuronal activity. However, experimentation aimed at optimal choice of parameters for such models is essential and results are reported in detail. Results derived from ensemble neuronal data revealed highly repeatable patterns of state transitions in the order of milliseconds in response to probing stimuli. PMID- 21622082 TI - Designing a thalamic somatosensory neural prosthesis: consistency and persistence of percepts evoked by electrical stimulation. AB - Intuitive somatosensory feedback is required for fine motor control. Here we explored whether thalamic electrical stimulation could provide the necessary durations and consistency of percepts for a human somatosensory neural prosthetic. Continuous and cycling high-frequency (185 Hz, 0.21 ms pulse duration charge balanced square wave) electrical pulses with the cycling patterns varying between 7% and 67% of duty cycle were applied in five patients with chronically implanted deep brain stimulators. Stimulation produced similar percepts to those elicited immediately after surgery. While consecutive continuous stimuli produced decreasing durations of sensation, the amplitude and type of percept did not change. Cycling stimulation with shorter duty cycles produced more persisting percepts. These features suggest that the thalamus could provide a site for stable and enduring sensations necessary for a long term somatosensory neural prosthesis. PMID- 21622083 TI - Impact of the improved patsari biomass stove on urinary polycyclic aromatic hydrocarbon biomarkers and carbon monoxide exposures in rural Mexican women. AB - BACKGROUND: Cooking with biomass fuels on open fires results in exposure to health-damaging pollutants such as carbon monoxide (CO), polycyclic aromatic hydrocarbons (PAHs), and particulate matter. OBJECTIVE: We compared CO exposures and urinary PAH biomarkers pre- and postintervention with an improved biomass stove, the Patsari stove. METHODS: In a subsample of 63 women participating in a randomized controlled trial in central Mexico, we measured personal CO exposure for 8 hr during the day using continuous monitors and passive samplers. In addition, first-morning urine samples obtained the next day were analyzed for monohydroxylated PAH metabolites by gas chromatography/isotope dilution/high resolution mass spectrometry. Exposure data were collected during the use of an open fire (preintervention) and after installation of the improved stove (postintervention) for 47 women, enabling paired comparisons. RESULTS: Median pre and postintervention values were 4 and 1 ppm for continuous personal CO and 3 and 1 ppm for passive sampler CO, respectively. Postintervention measurements indicated an average reduction of 42% for hydroxylated metabolites of naphthalene, fluorene, phenanthrene, and pyrene on a whole-weight concentration basis (micrograms per liter of urine), and a 34% reduction on a creatinine adjusted basis (micrograms per gram of creatinine). Pre- and postintervention geometric mean values for 1-hydroxypyrene were 3.2 and 2.0 MUg/g creatinine, respectively. CONCLUSION: Use of the Patsari stove significantly reduced CO and PAH exposures in women. However, levels of many PAH biomarkers remained higher than those reported among smokers. PMID- 21622084 TI - Maternal and gestational factors and micronucleus frequencies in umbilical blood: the NewGeneris Rhea cohort in Crete. AB - BACKGROUND: The use of cancer-related biomarkers in newborns has been very limited. OBJECTIVE: We investigated the formation of micronuclei (MN) in full term and preterm newborns and their mothers from the Rhea cohort (Crete), applying for the first time in cord blood a validated semiautomated analysis system, in both mono- and binucleated T lymphocytes. METHODS: We assessed MN frequencies in peripheral blood samples from the mothers and in umbilical cord blood samples. We calculated MN in mononucleated (MNMONO) and binucleated (MNBN) T lymphocytes and the cytokinesis block proliferation index (CBPI) in 251 newborns (224 full term) and 223 mothers, including 182 mother-child pairs. Demographic and lifestyle characteristics were collected. RESULTS: We observed significantly higher MNBN and CBPI levels in mothers than in newborns. In newborns, MNMONO and MNBN were correlated (r = 0.35, p < 0.001), and we found a moderate correlation between MNMONO in mothers and newborns (r = 0.26, p < 0.001). MNMONO frequencies in newborns were positively associated with the mother's body mass index and inversely associated with gestational age and mother's age, but we found no significant predictors of MNBN or CBPI in newborns. CONCLUSIONS: Although confirmation is needed by a larger study population, the results indicate the importance of taking into account both mono- and binucleated T lymphocytes for biomonitoring of newborns, because the first reflects damage expressed during in vivo cell division and accumulated in utero, and the latter includes additional damage expressed as MN during the in vitro culture step. PMID- 21622085 TI - Atrazine and cancer incidence among pesticide applicators in the agricultural health study (1994-2007). AB - BACKGROUND: Atrazine is a triazine herbicide used widely in the United States. Although it is an animal carcinogen, the mechanism in rodents does not appear to operate in humans. Few epidemiologic studies have provided evidence for an association. METHODS: The Agricultural Health Study (AHS) is a prospective cohort that includes 57,310 licensed pesticide applicators. In this report, we extend a previous AHS analysis of cancer risk associated with self-reported atrazine use with six additional years of follow-up and more than twice as many cancer cases. Using Poisson regression, we calculated relative risk estimates and 95% confidence intervals for lifetime use of atrazine and intensity-weighted lifetime days, which accounts for factors that impact exposure. RESULTS: Overall, 36,357 (68%) of applicators reported using atrazine, among whom there were 3,146 cancer cases. There was no increase among atrazine users in overall cancer risk or at most cancer sites in the higher exposure categories compared with the lowest. Based on 29 exposed cases of thyroid cancer, there was a statistically significant risk in the second and fourth quartiles of intensity-weighted lifetime days. There was a similar pattern for lifetime days, but neither the risk estimates nor the trend were statistically significant and for neither metric was the trend monotonic. CONCLUSIONS: Overall, there was no consistent evidence of an association between atrazine use and any cancer site. There was a suggestion of increased risk of thyroid cancer, but these results are based on relatively small numbers and minimal supporting evidence. PMID- 21622086 TI - New Jersey: a case study of the reduction in urban and suburban air pollution from the 1950s to 2010. AB - BACKGROUND: Air pollution has been a topic of intense concern and study for hundreds of years. During the second half of the 20th century, the United States implemented regulations and controls to reduce the levels of criteria air pollutants and achieve the National Ambient Air Quality Standards (NAAQS) for the protection of human health, while concurrently reducing the levels of toxic air pollutants. OBJECTIVE: In this commentary we trace the changes in air pollution in New Jersey as a case study, demonstrating the impact of local, state, and federal strategies to control emissions of pollutants and pollutant precursors from the 1950s until today. DISCUSSION: The original NAAQS (1970-1995) have been achieved, and significant progress has been made to achieve revised standards for ozone and particulate matter (PM) < 2.5 um in aerodynamic diameter (PM2.5) in New Jersey, which in the past was considered a highly polluted industrial state. CONCLUSIONS: Assuming no reversals on current regulations because of some major event or energy infrastructure disruption, outdoor air pollution reductions will continue to address health risks among specific segments of the general population affected by ozone/PM and pollution caused by neighborhood, local, and regional point and mobile sources. PMID- 21622087 TI - [MET: new target, new combinations]. AB - The MET tyrosine-kinase receptor is implicated in embryonic development and tissue repair. It appears to be a key of tumour development since it drives cell migration and invasion and can induce the conversion from an epithelial to a mesenchymal phenotype. Aberrant signaling of the MET pathways is associated with an aggressive prognosis and a poor outcome. Preliminary clinical results of several MET inhibitors have been encouraging particularly in tumours in which MET was amplified or mutated. MET inhibition could be especially interesting in association with others drugs since activation of MET is a secondary event induced by hypoxia, inflammatory cytokines or HER inhibitors that could exacerbate the malignant properties of transformed cells. Molecular targeted therapies against MET could therefore be effective as a combination approach. PMID- 21622088 TI - [New surgical procedures in musculoskeletal sarcomas of the child]. AB - The role of surgery remains central in the management of malignant bone tumors in children and adolescents. The current chemotherapies have demonstrated their efficiency (and limits) and made possible to avoid amputation in more than 90% of cases without worsening the prognosis. Advances in imaging (MRI) and understanding the mechanisms of tumor progression have led to more accurate resections with more frequent epiphyseal conservations. At the same time, new techniques have emerged that allows to consider more "organic" reconstructions. Therefore, we can better and more often preserve growth, hoping for a better long term functional outcome. Experience also allows to review older techniques (arthroplasties...). The choice of reconstruction depends on many factors (age, presence of secondary localizations, chemotherapy tolerance, socioeconomic context...) and can only be conceived as part of multidisciplinary evaluations. PMID- 21622089 TI - [Bronchial carcinoid tumors in children]. AB - Carcinoid tumors are the most common endobronchial tumors in the pediatric population, and represent a rare cause of airway obstruction. Clinical manifestations are unspecific, and diagnosis is often delayed due to low clinical suspicion. These tumors are considered low-grade malignant neoplasms, and their evolution is usually favorable after surgery. However, local recurrence and/or metastases can occur with both typical and atypical carcinoid tumors, justifying the need of prompt diagnosis and long-term follow-up. PMID- 21622090 TI - Giant Merkel cell carcinoma. PMID- 21622091 TI - Src-family tyrosine kinases as therapeutic targets in advanced cancer. AB - Src-family tyrosine kinases (SFK) play critical roles in mediating many cellular pathways such as proliferation, adhesion, survival, differentiation and cell motility. There is clear evidence that SFK activity is increased in many human cancers, either through gene amplification, transcriptional upregulation, posttranslational modification by activated upstream growth factor receptors, and even in rare cases, by mutations known to increase intrinsic tyrosine kinase activity in oncoviral forms of SFK. Many recent studies using animal models of human cancer seem to indicate that SFK may only be appropriate therapeutic targets in a subset of primary tumors because of the existence of multiple independent pathways that mediate oncogenic signaling. In contrast, SFK seem to be required for specific parameters of malignant progression, such as recurrence and/or metastasis- especially involving growth in the bone microenvironment. The resulting development of SFK antagonists, and their progression through clinical trials, has brought renewed focus on this tyrosine kinase family as critical mediators of the so-called lethal phenotype of cancer. PMID- 21622092 TI - Urban PM2.5 induces ROS generation and RBC damage in COPD patients. AB - Particulate matters (PM) produce adverse effects on the respiratory system and cause COPD. These effects are thought to involve intrinsic generation of ROS which are present in ambient PM (transition metals and aromatic organic compounds). Here, we examined the chemical composition and ultra-microscopic structure of PM2.5. The effect of this PM was studied in red blood cell (RBC) membranes (ghosts) from healthy volunteers (n = 11) and COPD patients (n = 43). These effects were compared with that produced by a Fenton metal-catalytic ROS generator. Oxidative biomarkers and cell damage were singificantly increased in presence of PM2.5 or ROS generator in RBC of COPD patients as compared with those in cells from healthy volunteers. In contrast, total SH groups, band 3 phospho tyrosine phosphatase (PTPase) and glucose-6 phosphate dehydrogenase (G6PD) activities were all diminished in cells from COPD patients. In conclusion, PM2.5 increases damage to RBCs from COPD patients, decreases the activity of PTPase and G6PD, and alters the function of the anionic exchanger (AE1) and the antioxidant response by decreasing SH groups. PMID- 21622093 TI - Effect of two pasteurization methods on the protein content of human milk. AB - The Holder method is the recommended pasteurization method for human milk banks, as it ensures the microbiological safety of human milk (HM). The loss of some biologically active milk components, due to the heat treatment, is a main limit to the diffusion of donor HM. High-temperature short-time (HTST) pasteurization may be an alternative to maintain the nutritional and immunological quality of HM. The aim of the present study was to compare the impact of Holder and HTST pasteurization on the HM protein profile. The protein patterns of HTST-treated milk and raw milk were similar. The Holder method modified bile salt-stimulated lipase, lactoferrin and components of the immune system. The HTST method preserved the integrity of bile salt-stimulated lipase, lactoferrin and, to some extent, of IgAs. Holder pasteurization decreased the amount of bile salt stimulated lipase and inactivated the remaining molecules, while the HTST method did not alter its activity. Pasteurization increased the bioavailable lysine quantity. HTST pasteurization seems to better retain the protein profile and some of the key active components of donor HM. PMID- 21622094 TI - Isolation and identification of novel geosmin-degrading bacteria. AB - Three novel geosmin-degrading bacteria were isolated from the sediments of Lake Kasumigaura, Japan. All strains were identified as Acinetobacter spp. by 16S rRNA gene sequence analysis and can biodegrade geosmin at an initial geosmin concentration of 2 mg/L after 2 days. Furthermore, at an initial geosmin concentration of 40 microg/L, geosmin removal was more than 68per cent by GSM-2 strain, and the degradation mechanism followed a pseudo-first-order mode. A rate constant of 0.026 reveals rapid geosmin degradation. This is the first report on geosmin degradation by by Acinetobacter spp. PMID- 21622095 TI - AIG1 is a novel Pirh2-interacting protein that activates the NFAT signaling pathway. AB - Pirh2 is an E3 ligase that negatively regulates p53 through both direct physical interaction and ubiquitin-mediated proteolysis. Here, we identified a novel Pirh2 interacting protein, AIG1, by yeast two-hybrid screening and confirmed its interaction with p53 both in vitro and in vivo. Quantitative real-time reverse transcription-PCR analysis showed that AIG1 expression levels were reduced in 50 out of 79 (63%) human hepatocellular carcinomas (HCCs) when compared to matched, non-cancerous liver tissue; levels were significantly different between HCCs with or without lymph node metastasis. Kaplan-Meier analysis indicated that the survival time of HCC patients down-regulated for AIG1 is much shorter than it is for patients up-regulated for AIG1 expression (p = 0.0313 as determined by the Log-rank test). Finally, AIG1 activated the nuclear factor of activated T cells (NFAT) signaling pathway in a dose-dependent manner when over-expressed in HEK293T cells. Our results suggest AIG1 could serve as a new biomarker for the diagnosis and prognostic evaluation of HCCs. PMID- 21622096 TI - Thyroid, spermatogenesis, and male infertility. AB - Since the identification of thyroid hormone receptors on the testes, thyroid has been suggested to have a significant impact on the male reproductive tract, spermatogenesis, and male fertility. Several research articles on the role of thyroid in spermatogenesis or male infertility have been published in the last three decades. We conducted an exhaustive literature search was conducted in order to create an up-to-date review of literature. This review aims to discuss the impact of thyroid on testicular development, spermatogenesis, hypo- or hyper- thyroidism and male infertility, and the management of thyroid related abnormal semen profile. The literature revealed that thyroid significantly impacts testicular development and that abnormal thyroid profile affects semen quality and male fertility by compromising testicular size, sperm motility and ejaculate volume. A clear link exists between thyroid hormones, testicular development and spermatogenesis. Thyroid disease negatively affects spermatogenesis and consequently may cause male infertility. In such cases, infertility is reversible, but more studies need to be conducted, especially in post-pubertal males to cement the current findings. PMID- 21622097 TI - Suppressing allergic immune responses. AB - While most studies are devoted to understanding the activation of cytokine gene transcription, few studies have focused on the silencing of cytokine gene transcription. This review will focus on the molecular mechanisms by which Th1 promoting factors downregulate the Il4 gene. PMID- 21622098 TI - Reference values of blood cell counts in the first days of life. AB - The lack of updated neonatal reference values for hematological parameters impacts significantly with clinical management of both healthy and sick newborns. The present pilot study was thus aimed at assessing updated hematological Italian reference values in late preterm and term newborns. From January 2004 to December 2008 hematological laboratory tests were performed in 1175 newborns (820 healthy and 355 sick controls) between 33-41 weeks of gestation, during the first four days after birth. Hematological parameters were sorted for gender and gestational age and statistically analyzed. No gender-related differences were observed at different weeks of gestation and no significant differences were found when study population was sub-grouped for late preterm and term newborns. During the first 4 days of life erythrocytes and platelets remained stable whilst white blood cell counts and differentials were significantly modified. This study shares updated reference values for hematological parameters in the early phases after birth and offers additional support for improving the management of sick infants. PMID- 21622099 TI - EGFR genomic alterations in cancer: prognostic and predictive values. AB - The role of EGFR in cancer development and progression has been recognized for long time in a variety of human malignancies including lung, head and neck, colon, breast, ovary and glioma. Recently its role as a target of antineoplastic agents has also been identified and a variety of EGFR-targeted drugs is already being used in a clinical setting and others are at present under investigation. Many data involving EGFR protein expression are now available for the choice of anti-EGFR monoclonal antibodies in colorectal cancer and with regard to EGFR gene mutations for the choice of tyrosine kinase inhibitors in lung cancer. Other EGFR related molecular factors, including the EGFR gene copy number, are currently under investigation. This review summarizes both preclinical and clinical available data regarding EGFR genomic alterations as prognostic and predictive factors. PMID- 21622100 TI - The roles of adenosine and adenosine receptors in bone remodeling. AB - Adenosine regulates a wide variety of physiological processes including heart rate, vasodilation and inflammation through the activation of specific cell surface adenosine receptors. In addition to these well-established roles of adenosine, recent genetic and pharmacological research has implicated adenosine as an important regulator in bone remodeling. The secretion of adenosine and the presence of its four receptors in bone cells have been well documented. More recently, we provided the first evidence that adenosine regulates osteoclast formation and function through A1 receptor (A1R), and showed that A1R-knockout mice have significantly increased bone volume as a result of impaired osteoclast mediated bone resorption. Moreover, adenosine A1R-knockout mice are protective from boss loss following ovariectomy further supporting the involvement of adenosine in osteoclast formation and function. This short review summarizes current knowledge related to the roles of adenosine and adenosine receptors in bone formation and remodeling. A deeper insight into the regulation of bone metabolism by adenosine receptors should assist in developing new therapies for osteoporosis. PMID- 21622101 TI - Introductory overview of purinergic signalling. AB - Purinergic neurotransmission was proposed in 1972 following identification of adenosine 5'-triphosphate (ATP) as the transmitter in non-adrenergic, non cholinergic inhibitory nerves in guinea-pig taenia coli. Subsequently ATP was identified as a co-transmitter in sympathetic, parasympathetic and most nerves in the peripheral and central nervous systems. ATP acts as a short-term signalling molecule in neurotransmission, neuromodulation and secretion and has long-term (trophic) roles in cell proliferation, differentiation and death in development and regeneration. Three subclasses of purine and pyrimidine receptors have been identified, P1 adenosine receptors (4 subtypes), P2X ionotropic nucleotide receptors (7 subtypes) and P2Y metabotropic receptors (8 subtypes). ATP is released physiologically by many cell types by mechanical deformation and, after release, ATP undergoes ectonucleotidase degradation. Purinergic receptors appeared early in evolution and have a widespread distribution on many non neuronal cells and neurons. Purinergic signalling is involved in embryonic and stem cell development. There is a rapidly growing literature about the pathophysiology of purinergic signalling including therapeutic developments for diseases, including stroke, thrombosis, osteoporosis, kidney failure, bladder incontinence, cystic fibrosis, dry eye, cancer and brain disorders. PMID- 21622102 TI - Regenerative medicine and tissue engineering in orthopaedic surgery. AB - Orthopedic surgery is going through a serious paradigm shift; instead of simply replacing damaged tissues with prosthetic or allograft material, the aim is to regenerate them. This endeavor has generated the field of regenerative orthopaedics, an increasingly expanding area of research with hopes of providing new and better treatments for diseases and injuries affecting the musculoskeletal system. As part of this process, we are witnessing a substantial accumulation of new cellular and molecular insights into connective tissue function, coupled with emerging new concepts in stem cell biology and scaffolding technologies. Indeed, any successful strategy to regenerate musculoskeletal tissues can be portrayed as an intricate interplay between the three main constituents of the regenerative system: cells, environment and scaffolds. This review is not meant to be exhaustive and comprehensive, but aims to highlight concepts and key advances in the field of regenerative orthopaedics and tissue engineering, as well as to present current possibilities for clinical translation. PMID- 21622103 TI - Inflammatory bowel disease in the geriatric population. AB - Inflammatory bowel disease (IBD) is a chronic disease that affects not only the young adults, but also the elderly. The elderly are more vulnerable and at higher risk from complications related to IBD. In this review we focus on IBD important features in the elderly and discuss the disease (1) epidemiology, (2) pathophysiology, (3) clinical manifestations and diagnosis, (4) prognosis, (6) therapy and (7) potential future research directions. PMID- 21622104 TI - Hypoxia-induced ROS signaling is required for LOX up-regulation in endothelial cells. AB - The adaptive response of endothelial cells to hypoxia involves a substantial remodeling of extracellular matrix (ECM). In endothelial cells hypoxia up regulates lysyl oxidase (LOX), a key enzyme in ECM assembly, relevant to vascular homeostasis. However, the mechanism underlying this response has not been established. Hypoxia up-regulated LOX expression in endothelial cells (HUVEC and BAEC) and concomitantly increased LOX enzymatic activity. This effect was independent of autocrine factors released by hypoxic cells and relies on a transcriptional mechanism. Both mTOR blockade and HIF-1alpha knockdown slightly prevented LOX up-regulation by hypoxia, suggesting that HIF-1alpha is only partially responsible for this effect. In fact, serial promoter deletion and mutagenesis studies indicated a limited contribution of the previously described hypoxia response element (-75 bp). Interestingly, Smad over-expression further increased LOX transcriptional activity in endothelial cells exposed to hypoxia. Moreover, the increase in LOX expression triggered by hypoxia was significantly reduced by reactive oxygen species (ROS) inhibitors. Thus, our data support a role of Smad signaling and ROS in the up-regulation of LOX by hypoxia in endothelial cells. PMID- 21622105 TI - C-Myb function in the vessel wall. AB - C-Myb is a DNA-binding transcription factor that functions in apoptosis, proliferation and differentiation. The role of c-Myb in vascular injury has been investigated previously both in vitro and in vivo, where knock-down of c-Myb is known to lead to a reduction in proliferation and an increase in apoptosis of vascular smooth muscle cells (VSMCs). Reduction of c-Myb activity has also been shown to decrease neointimal formation in vivo, by reducing VSMC proliferation. In contrast, over-expression of c-Myb in vivo leads to increased survival rates in certain cell types. This review will look mainly at studies investigating c Myb function in the vasculature, and evidence of signalling interactions which may be considered with regard to c-Myb as a possible target in the treatment of vasculoproliferative diseases. PMID- 21622106 TI - Sevoflurane preconditioning protects blood-brain-barrier against brain ischemia. AB - Sevoflurane preconditioning has recently been demonstrated to protect ischemic brain in vivo and in vitro. However, mechanisms underlying this neuroprotection have not been delineated. We therefore assessed the hypothesis that sevoflurane pretreatment protected blood-brain-barrier (BBB) via suppression of cell adhesion molecules (CAMs) and matrix metalloproteinases (MMPs) after ischemia. Repeated sevoflurane preconditioning was administered 24 hours before transient middle cerebral artery occlusion (MCAO). Neurologic deficits and expression of CAMs, MMPs and occludin were examined up to 3 days after ischemia. Evans blue (EB) extravasation and electron microscopy was detected at 2 days after ischemia. The data showed that sevoflurane pretreatment markedly improved BBB integrity and neurological outcomes after ischemia, robustly suppressed ischemia-induced decreases of occludin and increases of intracellular adhesion molecule-1 (ICAM 1), vascular cell adhesion molecule-1 (VCAM-1), MMP-2, MMP-9 and tissue inhibitor of metalloproteinase-1 (TIMP-1). Sevoflurane pretreatment also suppressed the activation of astrocytes and microglias in ipsilateral cortex and corpus callosum. In conclusion, repeated sevoflurane preconditioning confered potent protection against brain ischemia, partly by improving BBB integrity. PMID- 21622107 TI - B7-H3 and its relevance in cancer; immunological and non-immunological perspectives. AB - B7-H3 is a transmembrane glycoprotein and a member of the B7 family of proteins. It was previously known as an immunoregulatory molecule, shown in recent years to be of clinical significance in different types of cancer. In some tumor types high expression of B7-H3 has been linked to a poor prognosis, whereas in other cancers the opposite effect has been observed. Taken together, the precise role of B7-H3 in tumor immunity is unclear and further investigations are needed. Another aspect of B7-H3 that so far has received little interest is its role in non-immunological systems. We have demonstrated that knockdown of B7-H3 in melanoma and breast cancer cells results in both increased chemosensitivity and decreased metastatic potential. This has been observed in both in vitro and in vivo experiments. Several different signaling pathways seems to be involved, as B7-H3 knockdown can be linked to both higher expression of apoptotic markers and increased phosphorylation of Stat3. Increased knowledge of also the non immunological role of B7-H3 protein is therefore of great biological and putative therapeutic interest. PMID- 21622108 TI - Alterations of elastic fibers in genetically modified mice and human genetic diseases. AB - The two main components of elastic fibers are elastin and microfibrils. Fibrillin and microfibrillar-associated glycoproteins are the essential constituents of microfibrils. In the last ten years, the analysis of genetically modified mice and human genetic diseases has led to the observation that, besides elastin, fibrillins and microfibrillar-associated proteins, numerous other molecules play an essential role in the supramolecular organisation of the elastic fibers in the extracellular space and in the interactions between elastic fibers and cells. These recent data are summarized in this review. PMID- 21622109 TI - 1H NMR-based metabolomic analysis of urine from preterm and term neonates. AB - Metabolomics is a technique used to non-invasively determine metabolic status of an organism. Aim of our study was to analyze urinary metabolic profiles in term and preterm infants in order to identify gestational age-related metabolic differences and to predict metabolic maturity at birth. Twenty-six healthy term infants and 41 preterm infants were prospectively enrolled. A urine sample was collected non-invasively within the first hours of life. Samples were analyzed by proton nuclear magnetic resonance (1H NMR) spectroscopy and NMR urine spectra were analyzed by multivariate statistical analysis. Distinct metabolic patterns were found between term infants and preterm infants, as well as between preterm infants of 23-32 weeks' gestation and those of 33-36 weeks' gestation. Individual metabolites discriminating between these groups were hippurate, tryptophan, phenylalanine, malate, tyrosine, hydroxybutyrate, N-acetyl-glutamate, and proline. Metabolomic analysis revealed distinct urinary metabolic profiles in neonates of different gestational ages, and identified the discriminating metabolites. This holistic approach appears to be a promising tool for investigating newborn metabolic maturation over time, and might lead to a tailored management of neonatal disorders. PMID- 21622110 TI - Development of semiconductor nanomaterial whole cell imaging sensor on glass slides. AB - We report the development of a highly specific semiconductor quantum dots (QDs) based whole cell imaging sensor that offer rapid, reproducible, accurate, and long term cell imaging system on silanized microscope glass slides. The QD-based imaging sensor involved capture of whole cells with QD labeled highly specific antibodies against over expressed cell membrane proteins. The QDs were first modified with a polymer coating to generate carboxyl groups on the surface. Using the carboxylated QDs, antibodies were covalent conjugated using carbodiimide chemistry to form 2(0)Ab~QD that were used to capture whole cell. The SK-BR3 cell line was used as a model analyte in the sandwich type assay consisting of 1(0)Ab + SK-BR3 + Ab' + 2(0)Ab~QD. The assay was immobilized on an antibody modified silanized microscope slide that was subsequently mounted on a fluorescence microscope for detection. The results indicated that the QD based imaging sensor exhibited brighter signals compared with organic dye Texas red. The QD-based whole cell imaging sensor was visible under the microscope even after one week without fixation. PMID- 21622111 TI - Stage-specific mice ovarian somatic cell is involved in primordial folliculogenesis. AB - Although recent studies have shown that female germ cells can be produced from stem-cell lines in mice, whether these germ cells can interact with ovarian somatic cells (OSCs) to form primordial follicles (PFs) is still unclear. Here, we found after the PF pool is established, Irx3 and FoxL2 which were extensively expressed in the OSCs of the perinatal mouse decreased. Additionally, during primordial folliculogenesis, down-regulation of FoxL2 prevents OSCs invading germ cell cysts. Therefore, we investigated whether OSCs at different stages of folliculogenesis were able to reconstitute PFs with fetal ovarian germ cells in vitro. In this system, neither OSCs at the post-PF pool-establishment stage nor OSCs from normal or doxorubicin-treated adult mice were able to reconstitute PFs. Mice eliminated most of pre-existing PFs failed to exhibit any spontaneous regeneration of PFs. Our results suggest that OSCs can only support physiological primordial folliculogenesis during the prenatal period. OSCs lose the ability to form PFs after the establishment of the PF pool may be another reason why the PF pool is non-renewable in most adult mammals. PMID- 21622112 TI - Prune melanoidins protect against oxidative stress and endothelial cell death. AB - The health-promoting effects of fruit and vegetable consumption are thought to be due to phytochemicals contained in fresh plant material. Whether processed plant foods provide the same benefits as unprocessed ones is an open question. Melanoidins from heat-processed plums (prunes) were isolated and their presence confirmed by hydroxymethylfurfural content and browning index. Oxidative-induced endothelial cell (EC) damage is the trigger for the development of cardiovascular diseases (CVD); therefore the potential protective effect of prune melanoidins on hydrogen peroxide-induced oxidative cell damage was investigated on human endothelial ECV304 cells. Cytoplasmic and mitochondrial redox status was assessed by using the novel, redox-sensitive, ratiometric fluorescent protein sensor (roGFP), while mitochondrial membrane potential (MMP) was investigated with the fluorescent dye, JC-1. Treatment of ECV304 cells with hydrogen peroxide dose dependently induced both mitochondrial and cytoplasmic oxidation, in addition to MMP dissipation, with ensuing cell death. Pretreatment of ECV304 with prune melanoidins, significantly counteracted and ultimately abolished hydrogen peroxide elicited phenomena, clearly indicating that these polymers protect human EC against oxidative stress. PMID- 21622113 TI - Role of histamine H4 receptor in breast cancer cell proliferation. AB - In order to better understand the role of histamine H4 (H4R) receptor in breast cancer, we studied the receptor expression pattern, associated signal transduction pathway and biological responses, in breast cancer cell lines with different malignant characteristics. A different pattern of protein expression was observed in MDA-MB-231 compared to MCF-7 cells determined by western blot, exhibiting the presence of a diverse range of molecular weight species of the H4R. H4R agonist reduced cyclic adenosine monophosphate (cAMP) formation induced by forskolin only in MCF-7 cells. In MDA-MB-231 cells, H4R agonists significantly decreased cell proliferation, augmented the Annexin-V and TdT-mediated UTP-biotin Nick End labelling (TUNEL) positive cells and produced a 2.5-fold increase in cell senescence. In MCF-7 cells, H4R agonists inhibited proliferation by 50%, increasing the exponential doubling time. This effect was associated to an augment in Annexin-V and TUNEL positive cells, and a 2-fold increase in cell senescence. We conclude that H4R is functionally expressed in human breast cancer cell lines, exhibiting a key role in histamine-mediated biological processes such as cell proliferation, senescence and apoptosis. PMID- 21622114 TI - An introduction to quantum chemical methods applied to drug design. AB - The advent of molecular medicine allowed identifying the malfunctioning of subcellular processes as the source of many diseases. Since then, drugs are not only discovered, but actually designed to fulfill a precise task. Modern computational techniques, based on molecular modeling, play a relevant role both in target identification and drug lead development. By flanking and integrating standard experimental techniques, modeling has proven itself as a powerful tool across the drug design process. The success of computational methods depends on a balance between cost (computation time) and accuracy. Thus, the integration of innovative theories and more powerful hardware architectures allows molecular modeling to be used as a reliable tool for rationalizing the results of experiments and accelerating the development of new drug design strategies. We present an overview of the most common quantum chemistry computational approaches, providing for each one a general theoretical introduction to highlight limitations and strong points. We then discuss recent developments in software and hardware resources, which have allowed state-of-the-art of computational quantum chemistry to be applied to drug development. PMID- 21622115 TI - Costunolide inhibits proinflammatory cytokines and iNOS in activated murine BV2 microglia. AB - Costunolide, a sesquiterpene lactone present in Costus speciosus root exerts a variety of pharmacological activity but its effects on neuroinflammation have not been studied. Microglia, the resident phagocytic cells in the central nervous system respond to neuroinflammation and their overwhelming response in turn aggravate brain damage during infection, ischemia and neurodegenerative diseases. In this study, we report the effect of Costunolide on the production of proinflammatory mediators and mechanisms involved in BV2 microglial cells stimulated with LPS. Costunolide attenuated the expression of tumour necrosis factor-alpha, interleukin-1,6, inducible nitric oxide synthase, monocyte chemotactic protein 1 and cyclooxygenase 2 in activated microglia. This Costunolide-mediated inhibition was correspondent with the inhibition of NFkappaB activation. It has been further shown that Costunolide suppressed MAPK pathway activation by inducing MKP-1 production. Collectively our results suggest that Costunolide shows an ability to inhibit expression of multiple neuroinflammatory mediators and this is attributable to the compounds inhibition of NFkappaB and MAPK activation. This novel role of Costunolide upon investigation may aid in developing better therapeutic strategies for treatment of neuroinflammatory diseases. PMID- 21622116 TI - Non-coding RNAs and embryo implantation. AB - In mammals, thousands of non-protein-coding RNAs (ncRNAs), including microRNAs, endogenous small interfering RNAs, PIWI-interacting RNAs and mRNA-like long ncRNAs, have been identified. These RNAs modulate gene expression at transcriptional, post-transcriptional and epigenetic levels in many developmental and metabolic processes. Increasing evidence shows that ncRNAs are also linked to embryo implantation through regulating the expression of certain key genes and pathways. In this paper, we summarized the recent literatures on the ncRNAs involved in embryo implantation. PMID- 21622117 TI - Endothelial damage/dysfunction and hypertension in pregnancy. AB - Hypertension is one of the most common medical conditions complicating pregnancy, with significant implications on maternal and perinatal morbidity and mortality. Abnormalities in placentation have been implicated as the primary pathology responsible for the development of hypertension during pregnancy and its effects such as pre-eclampsia and eclampsia. With advancing research, the focus is now gradually shifting towards abnormalities in the maternal vasculature, including endothelial damage/dysfunction and impaired repair as a probable cause for this, with the latter being implicated in the development of cardiovascular disorders in later life in these women. Circulating endothelial cells (CECs) are a novel means of assessing endothelial dysfunction are mature cells detached from the vascular intimal layer in response to a variety of insults. Endothelial progenitor cells (EPCs) are non-leukocyte cells derived from the bone marrow with proliferative potential that may be important in vascular regeneration. This review article aims to provide an overview of current literature and concepts relating endothelial damage/dysfunction and impaired repair and the hypertensive disorders in pregnancy, with particular focus on CECs and EPCs. PMID- 21622118 TI - Nano-sized structures for the detection of food components and contaminants. AB - New methods to identify trace amount of food components and/or contaminants (infectious pathogens and chemicals) rapidly, accurately, and with high sensitivity are in constant demand to prevent foodborne illnesses. Multipurpose biofunctionalized engineered nanomaterials are very promising for the detection of food components and contaminants. The unique optical and magnetic properties of the nanoscale materials are very useful in the analysis of food. The objectives of this review paper are to discuss the development of applications of nanoscale structures related to food industries and to provide an overview of available methods of detecting food components and contaminants with particular emphasis on the use of nanoparticles. PMID- 21622119 TI - Postconditioning hormesis and the similia principle. AB - Postexposure conditioning, as a part of hormesis, involves the application of a low dose of stress following exposure to a severe stress condition. The beneficial effect of a low level of stress in postconditioning hormesis is illustrated by a number of examples found in experimental and clinical research. Depending on whether the low-dose stress is of the same type of stress or is different from the initial high-dose stress causing the diseased state, postconditioning is classified as homologous or heterologous, respectively. In clinical homeopathy, where substances are applied according to the Similia principle, the same distinction is found between the isopathic and the 'heteropathic' or homeopathic use of low dose substances. The Similia principle implies that substances causing symptoms in healthy biological systems can be used to treat similar symptoms in diseased biological systems. Only when heterologous substances are tested for therapeutic effects, the Similia principle can be studied. It is then possible to compare the effect of treatment with the degree of similarity between the diseased state and the effects caused by different substances. The latter research was mainly performed with cells in culture using heat shocked cells post exposed to a variety of stress conditions in low dose. PMID- 21622120 TI - DNA microarray analysis in endometriosis for development of more effective targeted therapies. AB - Microarray technology has become a widely used tool for analyzing the expression of tens of thousands of genes simultaneously on high-density microarrays in a single experiment, together with the availability of the complete nucleotide sequence of the human genome. DNA microarray technologies are powerful tools in the laboratory setting for scientific research, hypothesis generation, and the production of leads for subsequent validation through more sophisticated technologies. Since 2002, and particularly in the past few years, the utility of various DNA microarray technologies in endometriosis has rapidly and tremendously evolved; DNA microarray studies provide extremely important information that enable a better understanding of pathophysiology and disease etiology. This is a review of the literature focused on new findings of endometriosis pathophysiology obtained using various microarray technologies: gene expression profiling, microarray-based comparative genomic hybridization (CGH) (array CGH) single nucleotide polymorphism (SNP) arrays, the combination of chromatin immunoprecipitation (ChIP) with hybridization of microarrays (ChIP-on-chip). The present review discusses the results of a decade of DNA microarray technology experience, with emphasis on the pathophysiology of endometriosis in the context of clinical studies. PMID- 21622121 TI - TRAIL effect on osteoclast formation in physiological and pathological conditions. AB - Although osteoclasts (OCs) differentiate under the control of RANK/RANKL/OPG system, a number of inflammatory cytokines can contribute to increase osteoclastogenesis in diseases associated with bone loss. Recently, different studies indicate that TRAIL is implicated in modulating osteoclastogenesis. Here, we investigated the effect of TRAIL on OC formation in physiological and pathological conditions with bone involvement utilizing osteoclastogenesis in vitro models represented by peripheral blood mononuclear cells (PBMCs) from healthy donors and patients affected by multiple myeloma or periodontal disease. We demonstrated that in PBMCs from healthy donors TRAIL can directly induce OC formation in the absence of RANKL, while exert an inhibitory effect when added concomitantly to RANKL. In PBMCs from the patients, in which media the levels of TRAIL, RANKL and OPG are elevated, the neutralization of TRAIL partially inhibits the OC formation, and this effect was reversed by RANKL addition. Finally, we detect high TRAIL levels in the sera from the patients. In conclusion, our results indicate that TRAIL could exert a different role in modulating OC differentiation in physiological and pathological conditions. PMID- 21622122 TI - Gene expression profiles of in vivo derived mouse blastocysts after slow freezing. AB - The goals of our study were to analyze change of global gene expression profile of mouse blastocysts after slow-freezing and to explore molecular mechanism underlying the decreased pregnancy rate caused by cryopreservation. The results showed that superovulation differentially regulated the expression of 288 genes with at least 2.0-fold change considerations. Among which, 275 genes were down regulated and the remainder up-regulated in the cryopreserved group. The independent analysis with real-time PCR fully confirmed the results of microarray. These differential genes were classified into eighteen functional groups belonging to biological process, molecular function, and cellular component. Twenty nine genes could be categorized into one or more of KEGG pathways. The pathways of mitogen-activated protein kinase, Wnt and cell cycle were the most predominantly affected. Thus, the expression pattern reflected a broad spectrum of consequences of slow-freezing on the blastocysts, with most effects on stress-related and cell cycle-related genes. PMID- 21622123 TI - Principles, applications, risks and benefits of therapeutic hyperthermia. AB - Hyperthermia as a heat therapy is the procedure of raising the temperature of a part of or the whole body above normal for a certain period of time. Based largely on delivery methods, therapeutic hyperthermia falls under three major categories: local, regional, and whole-body. It may be applied alone or jointly with other modalities such as radiotherapy, chemotherapy, radiochemotherapy, and gene therapy. Because of the individual characteristics of each type of treatment, different types of heating systems have evolved. This paper provides an overview of possible mechanisms of heat-induced cell death and the way heating exerts its beneficial effect. It also discusses various heating devices as well as other modalities used with hyperthermia. The paper concludes with a summary of benefits and risks, obstacles encountered in the treatment process, and future research directions. PMID- 21622124 TI - Prognostic value of H-MLH1 after adjusting for RPA class in GBM patients. AB - Repair of DNA adducts appears to be an important mechanism in chemotherapy responsiveness in glioblastoma multiforme (GBM). Meta-analyses have suggested that the addition of chemotherapy increases the percentage of long-term survivors. Because GBM is characterized by multiplicity of pathways that characterize growth and treatment resistance, we hypothesized probing a multiplicity of repair factors may be able to identify more than one prognostic factor that may be utilized in molecularly targeted therapy that might improve survival and QOL. Seven DNA repair factors showed statistical significance when added to the initial logistic model of RPA class on length of survival status. After adjusting for RPA class the only statistically significant result of the multivariable logistic regressions for these 7 DNA repair factors was that as hMLH1-MF1 increased, the odds of being a short-term survivor versus a long-term survivor decreased (OR: 0.913, 95 per cent CI: 0.838-0.995, p=0.0385), multivariable analysis showed no associations between survival status and MGMT and p53 status, and the only statistically significant prognostic DNA repair factor was human Mut L Homologue 1 (hMLH1). PMID- 21622125 TI - Specific roles of threonine in intestinal mucosal integrity and barrier function. AB - Threonine is the second or third limiting amino acid in swine or poultry diets. This nutrient plays a critical role in the maintenance of intestinal mucosal integrity and barrier function, which can be indicated by intestinal morphology, mucus production (number of goblet cells), transepithelial permeability, brush border enzyme activity, and growth performance. Dietary threonine restriction may decrease the production of digestive enzymes and increase mucosal paracellular permeability. A large proportion of dietary threonine is utilized for intestinal mucosal protein synthesis, especially for mucin synthesis, and there is no oxidation of threonine by enterocytes. Because mucin proteins cannot be digested and reused, intestinal mucin secretion is a net loss of threonine from the body. Luminal threonine availability can influence synthesis of intestinal mucins and other proteins. Under pathological conditions, such as ileitis and sepsis, threonine requirement may be increased to maintain intestinal morphology and physiology. Collectively, knowledge about the role of threonine in mucin synthesis is critical for improving gut health under physiological and pathological conditions in animals and humans. PMID- 21622126 TI - Asthma treatment through the beta receptor: lessons from animal models. AB - Asthma is a significant health problem worldwide with a prevalence that continues to rise and for which there is no cure. Animal models have been used for decades to investigate the cause and cures of asthma, and while they do not always mimic many of the facets of this syndrome, mechanistic animal studies are still nevertheless very useful. Animal studies with beta-agonists suggest much broader and perhaps more important roles for beta-agonists since beta-agonists reduce aspects of inflammation and may affect structural remodeling. Studies using enantiomers of beta-agonists provide a confusing picture of the degree and mechanism of the deleterious effects of racemic mixtures and/or the S-enantiomer or other classes of beta-agonists. Neural mechanisms are implicated. The future holds a promise of even more insight into the mechanisms of the acute and chronic role of the beta-adrenoceptor, asthma therapeutics, in particular, beta-agonists that will lead to a better understanding of the pathogenesis and treatment of asthma. PMID- 21622127 TI - Developmental patterns of PPAR and RXR gene expression during spermatogenesis. AB - Members of the family of nuclear receptors that include peroxisome proliferator activated receptors (PPARs) and retinoid X receptors (RXRs) are important mediators of selective gene activation during development and cellular differentiation. In this study, developmentally-specific PPAR and RXR patterns of expression that occur in somatic and germ cell populations in the testis were determined using quantative real-time PCR (qRT-PCR) studies on RNAs that were isolated from StaPut purified mouse germ cells and primary rat Sertoli cells. These qRT-PCR studies indicate that transcripts encoding the PPAR-Alpha (alpha), Beta (beta), and -Gamma (gamma) and RXR -Alpha (alpha), -Beta (beta), and -Gamma (gamma) are developmentally expressed in both differentiating germ and Sertoli cells. In further experiments aimed at deciphering the physiological role that PPAR-Gamma (gamma) plays in Sertoli cells, 15-day primary rat Sertoli cells were infected with recombinant adenoviral vectors containing PPAR-Gamma (gamma) cDNA and PPAR-Gamma (gamma) RNAi constructs. Affymetrix microarray analysis and qRT PCR validation studies using total RNA isolated from these transfected cells indicated that PPAR-Gamma regulates the pattern of expression of key lipid metabolic genes in Sertoli cells. PMID- 21622128 TI - NADPH treatment decreases C6 glioma cell survival by increasing oxidative stress. AB - NADPH (nicotinamide adenine dinucleotide phosphate, reduced form) plays pivotal roles in antioxidation and reductive biosynthesis. However, the effect of NADPH treatment on cell survival is unknown. In this study, we determined the effect of NADPH treatment on the survival of glioma cells. Treatment of C6 glioma cells with as low as 1 MUM NADPH for 24 hrs induced a significant decrease in the survival of the glioma cells, while NADPH treatment had no effect on the survival of primary astrocyte cultures. We also found that NADPH treatment increased intracellular oxidative stress. Three antioxidants and the NADPH oxidase inhibitor, apocynin, attenuated the effect of NADPH. Poly(ADP-ribose) polymerase (PARP) activation appears to be a downstream effector of the oxidative stress, since PARP inhibitors reduced the effect of NADPH. Calcium chelator, BAPTA-AM, also attenuated the effect of NADPH. Collectively, these data indicate a novel property of NADPH: NADPH decreases glioma cell survival by inducing the NADPH oxidase-dependent increase in oxidative stress and by PARP activation. These results also suggest a potential therapeutic effect of NADPH on gliomas. PMID- 21622129 TI - Na+, K+-ATPase genes are down-regulated during adipose stem cell differentiation. AB - The expression of Na+, K+-ATPase alpha and beta subunits isoforms, FXYD2 and FXYD7 were studied in rat adipose stem cell (ASC) by qRT-PCR and immunofluorescence. ASCs were able to differentiate to chondrocytes or adipocytes. All studied genes were expressed in freshly isolated ASCs and in all passages checked. Immunostaining for alpha1 isoform was found in plasma membrane and nuclear envelope, alpha2 signal was lower and alpha3 staining was variable among cells. Beta isoforms signal was abundant and displayed an isoform-specific picture. Staining for FXYD7 was homogeneous in plasma membrane and cytosol. Chondrocytes differenciated from ASC showed identical Na+, K+-ATPase subunits isoforms expression patterns to chondrocytes in cartilage. The expression pattern of Na+, K+-ATPase genes in ASCs exhibits a unique phenotypic signature that implies functional differences in Na+ and K+ transport rates. Furthermore, this phenotypic signature may also be used as a complementary marker for studies of mesenchymal stem cell differentiation. We propose a possible 'moonlighting' role of Na+, K+-ATPase beta isoforms that could be essential for the study of mesenchymal stem cell function and differentiation. PMID- 21622130 TI - Involvement of LOC66273 isoform 2, a novel Mth938 containing protein, in MAPK pathway. AB - Using dual-luciferase reporter assay system, our previous study showed that LI2, significantly increased AP-1 transcriptional activity. Sub-cellular localization showed that GFP-LI2 fusion protein is diffusely distributed in the cytoplasm, with some highly concentrated spots around the nucleus, suggesting that LI2 protein has a physiological role in cytoplasm. Overexpression of LI2 significantly increased AP-1 transcriptional activity. Moreover, LI2 significantly promoted transcriptional activity of Elk1 and c-jun, which might, at least partly, be associated with activated ERK1/2 and JNK/SAPK signaling pathway. These data suggest that LI2 is a novel MAPK regulating protein. PMID- 21622131 TI - Novel molecular mechanisms by inorganic phosphate in osteosarcoma U2OS cells. AB - Osteosarcoma is the most common malignant primary bone tumor in children and adolescents and is characterized by a high metastatic potential. Its clinical outcome remains discouraging despite aggressive treatments. Thus, novel therapeutic approaches are needed. Recent results indicate that inorganic phosphate (Pi) is capable of affecting specific signal transduction pathways and of acting as an active regulator of cell behaviour. Previously, we found that Pi inhibits proliferation of human osteosarcoma U2OS cells via an adenylate cyclase/cAMP mediated mechanism. Here, we report that upon Pi treatment, U2OS cells become extremely hard to dislodge with trypsin. The lack of sensitivity to the trypsin action was paralleled by relevant changes in integrin subunits expression and accompanied by an increase of cell adhesion in cell-matrix adhesion assays. Interestingly, exposure of U2OS cells to Pi results also in a strong activation and protein level up-regulation of Rap1 small GTPase and in an early increase followed by a sustained inhibition of Erk1/2 phosphorylation. Importantly, the Pi-induced increase of cell adhesion was enforced by a cAMP analogue which specifically activated Epac/Rap1 and insensitive to PKA and MEK1/2 inhibitors. Our results enforce the evidences of inorganic phosphate as a signalling molecule, identify beta3 integrin, Rap1, ERK1/2 as proteins whose expression and function are relevantly affected by Pi in osteosarcoma U2OS cells The clinical significance and potential therapeutic applications by our data will be discussed. PMID- 21622132 TI - Optimal cut-off values for tumor markers in cerebrospinal fluid with ROC curve analysis. AB - To select optimal cut-off values of tumor markers in cerebrospinal fluid for the diagnosis of meningeal carcinomatosis, the concentrations of CEA, CA125, CA153, CA199, CA724, CYFRA21-1, AFP and NSE were determined by means of double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) in 21 MC patients, 25 cancer patients without leptomeningeal disease (group one) and 45 meningitis patients (group two) using the Roche E170 modular immunoassay analyzer. Optimal cut-off values were selected based on a receiver-operating characteristic curve. The results showed that CA125 and CEA in CSF were optimal diagnostic indices distinguishing between MC patients and cancer patients without leptomeningeal disease. Cut-off values for CA125 and CEA were 1.715 microliters and 0.274 microgram/l, respectively. In addition, CEA in CSF was the optimal diagnostic index distinguishing MC patients from meningitis patients. The cut-off value for CEA was 4.522 microgram/l. PMID- 21622133 TI - MicroRNA-210 as a novel blood biomarker in acute cerebral ischemia. AB - MicroRNA-210 (miR-210), a master and pleiotropic hypoxia-microRNA, plays multiple roles in brain ischemia. However, miR-210 expression and its function in humans have not been explored. The aim of our study is to evaluate the correlation of blood miR-210 with clinical findings in acute ischemic stroke. Blood samples were obtained from stroke patients (n=112) and healthy controls (n= 60). MiR-210 was measured at within 3, 7 and 14 days after stroke using a quantitative PCR technique. Stroke severity and clinical outcome were evaluated by NIHSS and modified Rankin Score. Both blood and brain miR-210 in ischemic mice was examined and the correlation was investigated. Compared to healthy controls, blood miRNA 210 was significantly decreased in stroke patients (0.93 vs. 1.36; P=0.001), especially at 7 days (0.56 vs. 1.36; P=0.001) and 14 days of stroke onset (0.50 vs. 1.36; P=0.001). The cut off point of miR-210 in diagnosis was 0.505 with 88.3 per cent sensitivity. MiR-210 level in stroke patients with good outcome was significantly higher than patients with poor outcome (1.2 vs. 0.44; P=0.012). The correlation between blood and brain miR-210 in ischemic mice was positive (R2=0.57, P=0.001). Blood miR-210 is a novel sensitive biomarker for clinical diagnosis and prognosis in acute cerebral ischemia. PMID- 21622134 TI - NPB001-05 inhibits Bcr-Abl kinase leading to apoptosis of imatinib-resistant cells. AB - The deregulated activity of the Bcr-Abl tyrosine kinase provides a rational basis for the development therapeutics in all phases of Chronic Myelogenous Leukemia (CML). Although a well studied imatinib therapy has clinical success against CML, resistance to imatinib due to mutations in the kinase domain, especially T315I poses a major problem for the ultimate success of CML therapy by this agent. Herein we describe an NPB001-05, derived from extract of Piper betle leafs, which is highly active in specifically inhibiting Bcr-Abl expressing cells. NPB001-05 inhibited the proliferation of BaF3 cells ectopically expressing wild type Bcr Abl phenotype and 12 different imatinib-resistant mutations of clinical relevance (average IC50 5.7 microg/ml). Moreover, NPB001-05 was highly inhibitory to wild type P210(Bcr-Abl) and P210(Bcr-Abl-T315I) kinase activity and abrogated the autophosphorylating enzyme in time- and dose- dependent manner. NPB001-05 was non toxic on normal cells, but was inhibitory to CML patient derived peripheral blood mononuclear cells. Treatment with NPB001-05 caused apoptosis induction and G0G1 cell cycle arrest in both Bcr-Abl wild type and T315I mutant cell lines. PMID- 21622135 TI - SNAT2 transceptor signalling via mTOR: a role in cell growth and proliferation? AB - We have investigated the effect of chronic competitive inhibition of SNAT2 (System A) amino acid (AA) transport, induced by incubation with a saturating dose of a non-metabolisable System A amino acid analogue (Me-AIB), on growth and proliferation of MCF-7 human breast cancer cells in complete culture medium. These cells express Na+- and pH-dependent SNAT2 AA transport and a saturating concentration of Me-AIB (10 mM) competitively inhibits (>90%) AA uptake via SNAT2. Incubation with Me-AIB for up to 5 days progressively reduced cell proliferation (~2-fold) and depleted intracellular concentrations of not only SNAT2 AA substrates but of essential branched chain AAs (e.g. leucine). Surprisingly, total cellular protein was maintained and cells subjected to chronic Me-AIB incubation exhibited a detectable increase in cell size. Analysis of mTOR signalling revealed that, despite a substantial reduction in size of the intracellular AA pool, Me-AIB elevated mTOR-dependent p70S6K1 phosphorylation. Proteomic analysis of TAP-tag purified SNAT2 fusion proteins identified two novel SNAT2-interacting proteins that may potentially function in conjunction with the SNAT2 transceptor to regulate signalling pathways influencing protein turnover and cell growth. PMID- 21622136 TI - Dispersion of Legionella-containing aerosols from a biological treatment plant, Norway. AB - Legionella was detected in aeration ponds (biological treatment plant) at Borregaard Ind. Ltd., Norway, and in air samples harvested directly above these ponds. Since 2005, three outbreaks of legionellosis occurred within a 10 km radius from this plant. This work addresses the dispersion patterns of Legionella containing particles by characterizing the aerosol plume emitted from these ponds (outbreak source) less than 500 meters using wind-tunnel measurements, CFD simulations, and real-life measurements. The most abundant particles directly over the ponds were less than 6 and more than 15 microm. The results showed that the aerosol plume remained narrow; 180 meters wide at 350 meters downwind of the ponds, and that 2 and 18 microm aerosols were mainly deposited in the vicinity of the ponds ( 150 - 200 meters). Furthermore, the maximum aerosol concentration level appeared 5-10 meters above ground level and the maximum concentration 500 meters downwind was approximately 2 per cent of the concentration level directly above the ponds. Our study demonstrates the strength of combining modeling with real-life aerosol analyses increasing the understanding of dispersion of airborne (pathogenic) microorganisms. PMID- 21622137 TI - The effect of pharmacological treatment on ADMA in patients with heart failure. AB - Asymmetric dimethylarginine (ADMA) plays a crucial role in the arginine-nitric oxide (NO) pathway. NO plays an important role in controlling vascular tone and regulates the contractile properties of cardiac myocytes. The purpose of this study was to investigate the effect of pharmacological treatment on asymmetrical dimethylarginine (ADMA) plasma levels in patients with acute congestive heart failure (HF). Patients with symptomatic acute congestive HF (NYHA Class III-IV) and impaired left ventricular (LV) function (ejection fraction less than 40 percent) were included in the study. ADMA and SDMA concentrations were assessed before and after pharmacological treatment in 18 critically ill patients on the intensive care unit by high performance liquid chromatography. All patients received a complete pharmacological treatment (diuretics, digoxin, ACE-inhibitors or angiotensin receptor blockers, and nitroglicerin) for the treatment of acute congestive HF. ADMA plasma levels of critically ill patients were significantly higher after pharmacological treatment respect baseline values (pre-treatment). In critically ill patients with acute congestive HF acute renal impairment function and the modulation of NOS determine plasma ADMA/SDMA levels after therapy. PMID- 21622138 TI - Sanggenon C decreases tumor cell viability associated with proteasome inhibition. AB - Several flavonoids have been reported to be proteasome inhibitors, but whether prenylated flavonoids are able to inhibit proteasome function remains unknown. We report for the first time that Sanggenon C, a natural prenylated flavonoid, inhibits tumor cellular proteasomal activity and cell viability. We found that (1) Sanggenon C inhibited tumor cell viability and induced cell cycle arrest at G0/G1 phase; (2) Sanggenon C inhibited the chymotrypsin-like activity of purified human 20S proteasome and 26S proteasome in H22 cell lysate, and Sanggenon C was able to dose-dependently accumulate ubiquitinated proteins and proteasome substrate protein p27; (3) Sanggenon C-induced proteasome inhibition occurred prior to cell death in murine H22 and P388 cell lines; (4) Sanggenon C induced death of human K562 cancer cells and primary cells isolated from leukemic patients. We conclude that Sanggenon C inhibits tumor cell viability via induction of cell cycle arrest and cell death, which is associated with its ability to inhibit the proteasome function and that proteasome inhibition by Sanggenon C at least partially contributes to the observed tumor cell growth inhibitory activity. PMID- 21622139 TI - Separation of anti-neoplastic activities by fractionation of a Pluchea odorata extract. AB - Natural products continue to represent the main source for therapeutics, and ethnopharmacological remedies from high biodiversity regions are a rich source for the development of novel drugs. Hence, in our attempt to find new anti neoplastic activities we focused on ethno-medicinal plants of the Maya, who live in the world's third richest area in vascular plant species. Pluchea odorata (Asteraceae) is traditionally used for the treatment of various inflammatory disorders and recently, the in vitro anti-cancer activities of different extracts of this plant were described. Here, we present the results of bioassay-guided fractionations of the dichloromethane extract of P. odorata that aimed to enrich the active principles. The separation resulted in fractions which showed the dissociation of two distinct anti-neoplastic mechanisms; firstly, a genotoxic effect that was accompanied by tubulin polymerization, cell cycle arrest, and apoptosis (fraction F2/11), and secondly, an effect that interfered with the orchestrated expression of Cyclin D1, Cdc25A, and Cdc2 and that also led to cell cycle arrest and apoptosis (fraction F3/4). Thus, the elimination of generally toxic properties and beyond that the development of active principles of P. odorata, which disturb cancer cell cycle progression, are of interest for potential future therapeutic concepts against proliferative diseases. PMID- 21622140 TI - Acylated catechin derivatives: inhibitors of DNA polymerase and angiogenesis. AB - Catechins in green tea display anti-cancer and anti-angiogenesis activities. We previously found that some catechins, such as epigallocatechin-3-O-gallate (EGCG), inhibit the activities of eukaryotic DNA polymerases (pols) (Y. Mizushina et al.: Structural analysis of catechin derivatives as mammalian DNA polymerase inhibitors. Biochem Biophys Res Commun 333, 101-109 (2005)). In this study, we discuss the effects of chemical modifications of catechin and epicatechin that enhance their anti-cancer and anti-angiogenic activities based on pol inhibition. Catechins conjugated with fatty acid (3-O-acylcatechins) are stronger inhibitors of mammalian pol than epicatechins conjugated with fatty acid (3-O acylepicatechins). Moreover, 3-O-acylcatechins are more potent inhibitors of cultured cell growth both of the human colon carcinoma cell line (HCT116 cells) and human umbilical vein endothelial cell (HUVEC) line, as well as angiogenesis by comparison with 3-O-acylepicatechins. Catechin conjugated with stearic acid ((2R,3S)-3',4',5,7-tetrahydroxyflavan-3-yl octadecanoate; C-C18) was the strongest inhibitor in replicative pol alpha and repair-related pol beta, as well as the cultured cell growth and angiogenesis assays in the compounds tested. C C18 also suppressed HUVEC tube formation on reconstituted basement membrane suggesting that it affected not only pols but also signal transduction pathways in HUVECs. These data indicate that the acylated catechins target both pols and angiogenesis as anti-cancer agents. Moreover, the results suggest that acylation of catechin is an effective chemical modification to improve the anti-cancer activity of catechin. PMID- 21622141 TI - Antitumor activity of NPB001-05, an orally active inhibitor of Bcr-Abl tyrosine kinase. AB - Scientists are constantly searching for phytochemical compounds with anti-cancer activity. In this study, activity of plant extract NPB001-05 from Piper betle was tested on human chronic myelogenous leukemia (CML) xenograft models. NPB001-05 was active when dosed orally (500 mg/kg) once or twice a day in xenograft tumor models. NPB001-05 showed activity to T315I tumor xenograft, where imatinib failed to show antitumor activity. NPB001-05 showed no relevant toxicity in animal models during 2 weeks exposure to drug. Responsive tumor showed inhibition of tyrosine kinase activity with lowered Bcr-Abl protein levels and increased apoptosis. Microarray based transcription profiling studies demonstrated that both imatinib and NPB001-05 dysregulated imatinib- responsive genes. NPB001-05 showed additional genes selectively dysregulated from ER stress, PI3K/AKT, MAPK pathways. Additionally, we tested gene expression of PI3K, AKT1, JUN, CASP3 and DDIT3 in K562, BaF3P210(BCR-ABL) and BaF3 P210(BCR-ABLT315I) cell line treated for 6- and 12- hours with NPB001-05 and imatinib. The data indicates that NPB001 05 mediated cell death in K562 affects the function of ER stress. NPB001-05 shows antitumor activity with favorable toxicity profile. PMID- 21622142 TI - Physiological normoxia and chondrogenic potential of chondrocytes. AB - Cartilage is poorly vascularised with a limited capacity for repair following damage. The poor vascularisation results in cartilage tissue having a low normoxic value. This study examined and compared the effects of physiological cartilage normoxia (2% O2), hypoxia (0.2% O2), and hyperoxia (21% O2) on human articular chondrocytes (hAC) during similar time courses to those prior to transplant in cell therapy procedures. hAC were isolated and maintained at 0.2% O2, 2% O2, or 21% O2. Population doublings (PDs), cell surface area, chondrogenic differentiation potential, RT-PCR, quantitative RT-PCR and immunohistochemistry (Collagen Type II) were used to confirm chondrogenic differentiation of micromass pellets in different O2. Isolation and maintenance of hAC at =2% O2 resulted in significant alterations in surface area (smaller), rate of proliferation (reduced), and chondrogenic differentiation potential (enhanced). Chondrogenic gene expression appeared largely insensitive to O2 concentration. A relationship was apparent between collagen type II protein presence and O2 concentration. Oxygen concentrations of 2% O2 or less promoted retention of a dedifferentiated hAC phenotype and enhanced stability of hAC chondrogenesis. PMID- 21622143 TI - Long-term phenylbutyrate administration prevents memory deficits in Tg2576 mice by decreasing Abeta. AB - Aberrations in protein folding, processing, and/or degradation are common features of neurodegenerative diseases, such as Alzheimer's disease (AD). Sodium 4-phenylbutyrate (PBA) is a well-known histone deacetylase inhibitor, which increases gene transcription of a number of genes, and also exerts neuroprotective effects. PBA acts as a chemical chaperone reducing the load of mutant or unfolded proteins during cellular stress. Previously, we reported that 5-week administration of PBA reinstated memory loss and dendritic spine densities in the Tg2576 mouse model of AD. In this study we reported that chronic administration of PBA, starting before the onset of disease symptoms (6 month old) prevents age-related memory deficits in Tg2576 mice. The amelioration of the memory impairment is associated to a decrease in amyloid beta pathology and the glial fibrillary acidic protein (GFAP), suggesting that inflammation was reduced in PBA-treated animals. Together, the beneficial effects of PBA make it a promising agent for the prevention of AD. PMID- 21622144 TI - Charting the peptide crossreactome between HIV-1 and the human proteome. AB - This paper defines potential peptide cross-reactivity between HIV-1 and the human host. Specifically, the amino acid primary sequence of HIV-1, isolate CDC-451, was analyzed for potential immunopathological relationships with the human proteome. The results revealed that: 1) HIV-1 shares 50 heptapeptides and three octapeptides with the human proteome; 2) 34 of the 50 shared heptapeptides are experimentally validated epitopes targeted by immune responses following HIV-1 infection; 3) the viral heptapeptide epitopes are present in human proteins that, when altered, are associated with disease characteristics of acquired immunodeficiency syndrome (AIDS) such as CD4+ cell loss, encephalopathy, schizophrenia, myopathy, cardiovascular disorders, hypertension, corneal diseases, diarrhea, lymphoma, and bladder cancer; 4) at the pentapeptide level, the viral-versus-human overlap is extensive (14,227 matches), with the viral pentapeptides disseminated throughout 10,312 human proteins. The findings are discussed in relationship to HIV-1 escape from immune surveillance, adjuvant induced HIV-1 immunogenicity, autoimmune cross-reactions following human hyperimmune responses against HIV-1, and AIDS. PMID- 21622145 TI - Interactions of thyroid hormone and FSH in the regulation of rat granulosa cell apoptosis. AB - Thyroid hormone (TH) is important for normal reproductive function. Our previous studies indicate that FSH increases preantral follicle growth in vitro, a response markedly enhanced by triiodothyronine (T3). However, the nature of this hormonal interaction is poorly understood. The objective of this study was to determine if and how T3 modulate FSH-induced expression and actions of granulosa cell intracellular survival and death intermediates. We investigated the possible involvement of Src and PI3K/Akt pathway in the regulation of granulosa cell survival. We demonstrated that, while ineffective alone (0.1-100 nM), T3 markedly enhanced FSH (100 ng/ml)-induced granulosa cell phospho-Src and phospho-Akt contents and Xiap expression in vitro. The effects of T3 were concentration dependent, with maximal responses at 1.0 nM. FSH alone decreased Fas Ligand (FasL) content irrespective of the presence of T3. Co-treatment of cell with T3 and FSH decreased Fas content, although neither hormone alone elicited a significant response. Taken together, the present study demonstrates that T3 potentiates the cell survival action of FSH through Src- and PI3K-mediated Xiap up-regulation and decreased Fas and FasL expression. PMID- 21622146 TI - Histamine regulates relevant murine dendritic cell functions via H4 receptor. AB - Histamine, produced by dendritic cells (DCs) or by other cells of the immune system, may have significant impact on DC activities. We investigated the influence of histamine and histamine H4 receptor (H4R) on some relevant functions of DCs. Histamine significantly decreased the antigen presentation capacity of splenic DCs, and this effect was reversed by a H4R antagonist. Furthermore, enhanced antigen presentation was detected in H4R-/- DCs. Prolonged histamine treatment during DC differentiation stimulated migration, albeit the increase was not significant. H4R-deficient DCs possessed significantly lower migration capacity than their wild-type counterparts. Monitoring in vivo and in vitro DC cytokine production revealed that a H4R agonist in combination with LPS, increased IL-1 beta mRNA expression, and a H4R antagonist reversed this effect. In H4R-deficient mice we detected decreased mRNA expression of some DC-derived cytokines including IFN-gamma and IL-10. Upon CFA stimulation, genotype-dependent differences were found in the expression of IL-6 and IFN-gamma. Our data suggest that H4R plays a crucial role in variety of functions of murine DCs. PMID- 21622148 TI - ST6GalNAc-I controls expression of sialyl-Tn antigen in gastrointestinal tissues. AB - Sialyl-Tn is a simple mucin-type carbohydrate antigen aberrantly expressed in gastrointestinal adenocarcinomas and in the precursor lesion intestinal metaplasia. Sialyl-Tn tumour expression is an independent indicator of poor prognosis. We have previously shown in vitro that ST6GalNAc-I and ST6GalNAc-II sialyltransferases can synthesize sialyl-Tn. The aim of the present study was to establish whether ST6GalNAc-I is the major enzyme responsible for the expression of sialyl-Tn. We used a model of CHO-ldlD cells producing only MUC1-Tn glycoform and showed that ST6GalNAc-I is the key-enzyme leading to sialyl-Tn biosynthesis. We developed novel monoclonal antibodies specific for ST6GalNAc-I and evaluated its expression in gastrointestinal tissues. ST6GalNAc-I was detected in normal colon mucosa co-localized with O-acetylated sialyl-Tn. Expression was largely unaltered in colorectal adenocarcinomas. In contrast, we found that ST6GalNAc-I is weakly expressed in normal gastric mucosa, but over-expressed in intestinal metaplasia, co-localized with sialyl-Tn. In gastric carcinomas ST6GalNAc-I was also associated with sialyl-Tn, but with heterogeneous staining and partial co localization. Our results showed ST6GalNAc-I as the major enzyme controlling the expression of cancer-associated sialyl-Tn antigen in gastrointestinal tissues. PMID- 21622147 TI - Intercalated disc-associated protein, mXin-alpha, influences surface expression of ITO currents in ventricular myocytes. AB - Mouse Xin-alpha (mXin-alpha) encodes a Xin repeat-containing, actin-binding protein localized to the intercalated disc (ICD). Ablation of mXin-alpha progressively leads to disrupted ICD structure, cardiac hypertrophy and cardiomyopathy with conduction defects during adulthood. Such conduction defects could be due to ICD structural defects and/or cell electrophysiological property changes. Here, we showed that despite the normal ICD structure, juvenile mXina null cardiomyocytes (from 3~4-week-old mice) exhibited a significant reduction in the transient outward K+ current (ITO), similar to adult mutant cells. Juvenile but not adult mutant cardiomyocytes also had a significant reduction in the delayed rectifier K+ current. In contrast, the mutant adult ventricular myocytes had a significant reduction in the inward rectifier K+ current (IK1) on hyperpolarization. These together could account for the prolongation of action potential duration (APD) and the ease of developing early afterdepolarization observed in juvenile mXin-alpha-null cells. Interestingly, juvenile mXin-alpha null cardiomyocytes had a notable decrease in the amplitude of intracellular Ca2+ transient and no change in the L-type Ca2+ current, suggesting that the prolonged APD did not promote an increase in intracellular Ca2+ for cardiac hypertrophy. Juvenile mXin-alpha-null ventricles had reduced levels of membrane-associated Kv channel interacting protein 2, an auxiliary subunit of ITO, and filamin, an actin cross-linking protein. We further showed that mXin-alpha interacted with both proteins, providing a novel mechanism for ITO surface expression. PMID- 21622149 TI - CD155 is involved in NK-cell mediated lysis of human hepatoblastoma in vitro. AB - NK cells are involved in the lysis of different solid tumors and leukemias. NK activity is thereby regulated by activating and inhibitory receptors. Until now, nothing is known about the NK-activity against hepatoblastoma and the involved receptors. We tested NK cells for cytotoxicity against HB in vitro. Expression levels of activating NK ligands were analysed on 13 primary HB samples as well as on 3 HB cell lines. All HB cell lines showed low HLA-class-I-expression. CD155 expression was strong on primary HB samples and cell lines. NKG2D-ligands (MICA/B, ULBP1-3) were heterogeneous expressed in primary samples and cell cultures. There were no differences between the various histological subtypes. NK cells showed strong cytotoxicity in vitro which was significantly increased through interleukin-2 and -15 stimulation (p less than 0.001). Blockade of CD155 resulted in decreased lysis rates. Our findings show that NK cells exert high activity against hepatoblastoma in vitro and that CD155 is involved in the NK mediated killing of HB. The inclusion of a NK-based immunotherapy into novel treatment strategies might be a promising alternative especially for advanced tumors. PMID- 21622150 TI - The A640G CYBA polymorphism associates with subclinical atherosclerosis in diabetes. AB - Oxidative stress is implicated in diabetes. The NADPH oxidases are the main source of superoxide in phagocytic and vascular cells, and p22phox is a key subunit. Genetic variants of CYBA, the human p22phox gene, associate with cardiovascular disease. We investigated the association of the A640G polymorphism with diabetes and its impact on phagocytic NADPH oxidase-dependent superoxide production and subclinical atherosclerosis. We studied 1212 subjects in which clinical parameters including carotid intima-media thickness (cIMT) were assessed. The A640G polymorphism was genotyped by TaqMan probes. In 496 subjects, the NADPH oxidase-dependent superoxide production in peripheral blood mononuclear cells was assessed by chemiluminescence. The GG genotype prevalence was significantly higher in type 2 diabetic patients than in non-diabetic subjects. Peripheral blood mononuclear cells from diabetic GG patients presented higher NADPH oxidase-dependent superoxide production than those of diabetic AA/AG patients. Within the diabetic group, GG patients presented higher cIMT levels than AA/AG patients. The A640G CYBA polymorphism may be a marker of oxidative stress risk and may be indicative of subclinical atherosclerosis in type 2 diabetes. PMID- 21622151 TI - Additional 5-FU-LV significantly increases survival in gastrointestinal cancer. AB - Metastatic colorectal and other locally advanced gastrointestinal (G.I.) cancers often recur after curative resection. Many mechanisms of tumor growth and/or immune escape by residual cancer cells may provoke tumor progression. Long-term, cytostatic action with repeated post-adjuvant administration of 5-fluorouracil (FU)-leucovorin (LV) cycles may interrupt or downregulate these mechanisms and favor the recovery and/or increase the immune system activity. Seventy patients were considered. An active prospective cohort including 21 patients (study group) was matched in a 1:1 ratio with a retrospective parallel control group of 21 patients. The study group received long-term repeated post-adjuvant administration of 5-FU-LV cycles, while the matched control group was conventionally treated. Statistical analysis was performed by Kaplan-Meier method and Cox's proportional hazard regression model. The five-year disease-free survival (DFS) was 77.0 + 10.1 % and 31.7 + 10.6 % (p = 0.001; hazard ratio (HR) 5.3, 95 % C.I.: 1.7-16.1, p = 0.003), while the five-year overall survival (OS) was 88.0 + 8.1 % and 37.0 + 10.7 % (p = 0.001; HR 8.9, 95 % C.I.: 2.0-39.9, p = 0.004) in the study group and in matched controls respectively. These findings suggest a relevant improvement in the outcome of this population by an intermittent and prolonged cytostatic effect with 5-FU-LV. PMID- 21622152 TI - Garcinol-induced apoptosis in prostate and pancreatic cancer cells is mediated by NF- kappaB signaling. AB - Garcinol, obtained from Garcinia indica, is a potent antioxidant. Its anticancer activity has been investigated; however, there is no published report on its action against prostate and pancreatic cancer cells. We have earlier reported its activity against breast cancer cells, and here we tested our hypothesis that garcinol could inhibit cell proliferation and induce apoptosis in prostate as well as pancreatic cancer cells. Using multiple techniques such as MTT, Histone DNA ELISA, activated caspase assays, clonogenic assays and EMSA, we investigated the mechanism of apoptosis-inducing effect of garcinol in prostate (LNCaP, C4-2B and PC3) and pancreatic (BxPC-3) cancer cells. We found that garcinol inhibited cell growth of all the cell lines tested with a concomitant induction of apoptosis in a dose-dependent manner. Down-regulation of NF-kappaB signaling pathway appears to be the mechanism of apoptosis-induction because garcinol inhibited constitutive levels of NF-betaB activity, which was consistent with down-regulation of NF-betaB-regulated genes. A significant decrease in the colony forming ability of all the cell lines was also observed, suggesting the possible application of this compound against metastatic disease. In summary, our results provide pre-clinical evidence to support the use of garcinol against human prostate and pancreatic cancer, thus meriting its further investigation as a potential chemo-preventive and/or therapeutic agent. PMID- 21622153 TI - Identification of protein-protein interactions of human HtrA1. AB - The human heat shock protein HtrA1, a member of the HtrA family of serine proteases, is a evolutionarily highly conserved factor which displays a widespread pattern of expression. The yeast two-hybrid technique was employed to identify new cellular proteins physically interacting with HtrA1, and thus potential targets of this serine protease. An enzymatically inactive HtrA1 point mutant, HtrA1-S328A, was generated and used as bait in a yeast two-hybrid system. Fifty-two plasmids were isolated from primary positive yeast clones. Subsequent sequencing and BLAST analysis revealed cDNAs encoding for 13 different proteins. These putative binding partners of HtrA1 appeared to be a) components of extracellular matrix; b) factors related to signal pathways, and c) unknown proteins. Among the 13 positive clones identified and reported here, it is worth of note that the interaction of HtrA1 with tubulin and collagen (extracellular matrix proteins) and with tuberin (cytoplasmic protein) is confirmed by other studies, and this further supports previous findings in which HtrA1 can be found active as an intracytoplasmic protein or as secreted protein as well. PMID- 21622154 TI - Novel mobilization strategies to enhance autologous immune effector cells in multiple myeloma. AB - The immune system plays a critical role determining the outcomes in transplanted multiple myeloma patients, since enhanced lymphocyte recovery results in improved survival. Since mobilization regimens influence the cellular subsets collected and infused for transplant, these regimens may determine immune recovery following transplant. We hypothesized that a mobilized stem cell product harboring an increased number of lymphocytes would enhance immune recovery following autologous stem cell infusion, increase lymphocyte recovery, and improve clinical outcomes. We designed a phase I immune mobilization trial using IL-2 and growth factors to increase the number of lymphocytes within the stem cell product. This regimen efficiently mobilized CD34+ progenitor cells (median: 3.6 x 10(6) cells/kg; range 1.9-6.6 x 10(6) cells/kg) and improved the immune properties of the mobilized stem cells, including an increase in CD8+ T cells expressing an NK activating receptor called NKG2D (P less than 0.004), cells that are extremely potent at killing myeloma cells using non-MHC-I restricted and TCR independent mechanisms. Novel mobilization techniques can improve the mobilized graft and may improve clinical outcomes in myeloma patients. PMID- 21622155 TI - Resveratrol potentiates grape seed extract induced human colon cancer cell apoptosis. AB - Colon cancer is the third leading cause of cancer deaths in men and women. Grape seed extract (GSE) and resveratrol (RSV) are potent chemopreventive agents against colon cancer both in vitro and in vivo, at relatively high concentrations. We hypothesized that RSV and GSE may act in concert with each other in potentiating their anti-cancer properties at sub-optimal doses, because they occur as complex mixtures in grapes. In this study, we showed that RSV (~25 micromolar) potentiated GSE (<= 35 microg/mL) induced colon cancer cell apoptosis via activation of p53 dependent pathways. Elevation of apoptosis was much more pronounced in p53 +/+ cells compared to p53 -/- cells. Apoptosis was strongly correlated with pp53 levels and Bax:Bcl-2 ratio, key players in the mitochondrial apoptotic pathway. Caspase-3 inhibition and reactive oxygen species suppression attenuated apoptosis induced by the combination. RSV-GSE combination suppressed proliferation and induced apoptosis even in the presence of mitogenic growth factor IGF-1, suggesting the importance of understanding the potentiating effects of phytonutrients in combination as they would occur in nature rather than individually. PMID- 21622156 TI - Oxidative stress in primary open-angle glaucoma. AB - Glaucoma is the second leading cause of blindness worldwide and is still a mysterium. Despite some risk factors are known, like individually elevated intraocular pressure, myopia, age, genetic factors, as well as vascular risk factors and smoking, the exact mechanism developing a glaucomatous optic neuropathy are still unknown. In the pathogenesis of glaucoma oxidative stress seems to play an important role. The mitochondria have an abnormal DNS and the antioxidative capacity is reduced. In addition, in the anterior segment, e.g. trabecular meshwork as well as in the posterior pole glaucoma patients show an increased oxidative stress. Therefore oxidative stress should be considered in therapeutic approaches to glaucoma patients. PMID- 21622157 TI - Genistein stimulates the insulin-dependent signaling pathway. AB - Small compounds that activate the insulin-dependent signaling pathway have potential therapeutic applications in controlling insulin-independent diabetes mellitus. In this study, we investigated whether soybean isoflavones could induce the expression of SHARP-2, a downstream component of insulin-dependent signaling pathway, associated with the regulation of blood glucose. One such compound called genistein, rapidly and temporarily induced SHARP-2 mRNA levels in a dose dependent manner in rat H4IIE hepatoma cells. This induction process was rapidly stimulated by a protein kinase C (PKC) activator and blocked by a PKC inhibitor, suggesting that SHARP-2 may be induced via PKC activation. Upon Western blot analysis, genistein showed a stimulation of PKC phosphorylation. Therefore, we concluded that genistein might transcriptionally induce SHARP-2 through the activation of PKC in H4IIE cells. Our results suggest that genistein might be a useful dietary supplement to control insulin-independent diabetes mellitus by inducing the SHARP-2 expression via a bypass of the insulin-dependent signaling pathway. PMID- 21622158 TI - EPO reduces reactive gliosis and stimulates neurotrophin expression in Muller cells. AB - To characterize Muller cell-mediated neuroprotective and neurotrophic functions of the erythropoietin (EPO)/EPO receptor (EpoR) system in diabetic rat retina. A single intravitreal injection of EPO (8 mU/eye) was administered in rats 4 or 24 weeks after diabetes onset. The results showed that intravitreal EPO ameliorated the up-regulation of GFAP and vimentin in the diabetic retina evaluated by immunofluorescence and Western blotting; but up-regulated BDNF and CNTF expressions, quantified by real-time PCR and ELISA, in the 24-week diabetic rat retinas. In vitro, BDNF and CNTF expressions were stimulated by EPO through both extracellular signal-regulated kinase1/2 (ERK1/2) and Akt pathways. The neuro regenerative function of EPO, as indicated by promotion of neurite outgrowth, was corroborated in vitro. BDNF was involved in EPO-induced neurite outgrowth of primary rat retinal neurons. Exogenous EPO exerts neuroprotective and neurotrophic functions by attenuating reactive gliosis and promoting neurotrophic factors in Muller cells in diabetic retina. Signaling pathways that are responsible for these Muller cell-mediated EPO/EpoR functions may be therapeutic targets for diabetic retinopathy. PMID- 21622159 TI - Assessment of tumor response to tyrosine kinase inhibitors. AB - This review briefly summarizes recent developments in the use of non-invasive imaging to assess tumor response to TKI therapy. Receptor tyrosine kinases play important roles in cancer development. A new class of drugs, tyrosine kinase inhibitors (TKI) can induce rapid and dramatic tumor suppression when administered to carefully selected patient groups. Identifying these patients with responding tumors prior to or shortly after the initiation of therapy remains challenging. The gold standard of response assessment has been by invasive biopsies used in biological and biochemical procedures. Advances in non invasive imaging at the anatomical, functional and molecular level have enabled the early detection of tumor response; sometimes within days of beginning treatment. The growing area of molecular imaging has spurred the discovery of novel targeting peptides to bind TKI responding tumors. The emergence of targeted, quick responding imaging probes advances the field of cancer management towards the goal of personalized medicine. PMID- 21622161 TI - Confirmation and further mapping of the GLC3C locus in primary congenital glaucoma. AB - We investigated the relationship between primary congenital glaucoma (PCG) in the Chinese Han population and its candidate locus GLC3C. 152 nuclear families (patients with normal parents) without carrying the CYP1B1 mutation were enrolled. Fluorescence Labeled Multiplex-PCR was used to genotype 12 short tandem repeats (STRs) within GLC3C region and transmission disequilibrium test (TDT) was used to analyze the association between PCG and these STR markers. Sixteen haplotype tag single nucleotide polymorphisms (htSNPs) were chosen from the location where the TDT tests showed positive results. Matrix-assisted laser desorption/ionization Time-of-flight (MALDI-TOF) mass spectrometry was used to perform SNP genotyping Haplotypes constructed from these SNPs were analyzed. The TDT results of STRs in the GLC3C area indicated that D14S279, D14S555 and D14S74 have significant transmission disequilibrium signals (p=0.0210, 0.0096 and 0.0034), with a genetic distance of 0.006 cM among them. Significant transmission disequilibrium (P=0.0010) occurred between the haplotype TAACG of rs2111701- rs4020123- rs4903696- rs11159318- rs177216 and the disease. Detection of disease causing genes within this region needs further study. PMID- 21622162 TI - Deciphering the function of nucleic acid sensing TLRs one regulatory step at a time. AB - While initial studies of Toll-like Receptor (TLR) signaling mainly focused on genetic analysis of signal transduction, recent work has highlighted the importance of understanding the basic cell biology underlying receptor function. Nowhere is this issue more important than in the study of the nucleic acid sensing TLRs. These receptors face the unique challenge of distinguishing microbial nucleic acids from similar host-derived molecules. The physiological cost of not making this distinction can be readily observed in studies of autoimmunity, a cause of which is often the inappropriate detection of self nucleic acids. In this review, we highlight recent research that has revealed myriad ways in which mammalian cells control the function of nucleic acid-sensing TLRs. A theme is now emerging whereby these receptors are subject to sequential regulatory mechanisms that control protein transport to their sites of signal transduction, as well as their access microbial nucleic acids. PMID- 21622163 TI - Lights and shadows on gene organization and regulation of gene expression in Leishmania. AB - Regulation of gene expression is one of the most intriguing aspects of Leishmania biology. This review deals with current knowledge concerning gene organization and regulation of gene expression in this protozoan parasite, which cause serious illness and death in humans living in tropical and subtropical regions. Post transcriptional regulation is especially important for Leishmania, and other trypanosomatids, due to the unusual features related to transcription and mRNA maturation. In these organisms, genes are organized into polycistronic transcription units, whereby many genes are cotranscribed by RNA polymerase II from not well characterized, upstream promoters. These organisms represent an extreme in which the expression of their genome is almost exclusively controlled post-transcriptionally. Because the regulatory needs of these parasites are considerable as they undergo complex developmental transitions, post transcriptional mechanisms that involve RNA and protein regulatory processes are of paramount importance for these protozoa. This review summarizes recent results on the post-transcriptional mechanisms in Leishmania that regulate protein abundance through influencing RNA splicing, nuclear-cytoplasmic mRNA stability, translation, or post-translational events such as protein stability and modification. PMID- 21622164 TI - Canonical and variant histones of protozoan parasites. AB - Protozoan parasites have tremendously diverse lifestyles that require adaptation to a remarkable assortment of different environmental conditions. In order to complete their life cycles, protozoan parasites rely on fine-tuning gene expression. In general, protozoa use novel regulatory elements, transcription factors, and epigenetic mechanisms to regulate their transcriptomes. One of the most surprising findings includes the nature of their histones--these primitive eukaryotes lack some histones yet harbor novel histone variants of unknown function. In this review, we describe the histone components of different protozoan parasites based on literature and database searching. We summarize the key discoveries regarding histones and histone variants and their impact on chromatin regulation in protozoan parasites. In addition, we list histone genes IDs, sequences, and genomic localization of several protozoan parasites and Microsporidia histones, obtained from a thorough search of genome databases. We then compare these findings with those observed in higher eukaryotes, allowing us to highlight some novel aspects of epigenetic regulation in protists and to propose questions to be addressed in the upcoming years. PMID- 21622165 TI - Functional characterization of MIMP for its adhesion to the intestinal epithelium. AB - The micro integral membrane protein (MIMP), the domain within the integral membrane protein of Lactobacillus plantarum CGMCC 1258, has been shown to adhere to mucin and antagonize the adhesion of enteroinvasive E. coli and enteropathogenic E. coli. To further characterize the functions of MIMP, we investigated its effects on the intestinal permeability, expression of tight junction (TJ) proteins and TJ ultrastructure in vitro and in vivo. We also determined the interaction between MIMP and dendritic cells (DCs). We observed that MIMP reduced intestinal permeability and restored the expression and distribution of TJ proteins in both NCM460 cell monolayers and in IL-10(-/-) mice. MIMP adhered to immature (i) DCs by binding to DC-SIGN, and induced DCs to produce anti-inflammatory cytokines and to mediate Th2 differentiation. Moreover, MIMP stimulated the expression of anti-inflammatory cytokines in colonic mucosa and attenuated colitis in IL-10(-/-) mice. In conclusion, MIMP is the main functional component of L. plantarum that contributes to its protective effects, and thus may be a potential therapeutic agent for intestinal diseases. PMID- 21622160 TI - Parallel evolution of nitric oxide signaling: diversity of synthesis and memory pathways. AB - The origin of NO signaling can be traceable back to the origin of life with the large scale of parallel evolution of NO synthases (NOSs). Inducible-like NOSs may be the most basal prototype of all NOSs and that neuronal-like NOS might have evolved several times from this prototype. Other enzymatic and non-enzymatic pathways for NO synthesis have been discovered using reduction of nitrites, an alternative source of NO. Diverse synthetic mechanisms can co-exist within the same cell providing a complex NO-oxygen microenvironment tightly coupled with cellular energetics. The dissection of multiple sources of NO formation is crucial in analysis of complex biological processes such as neuronal integration and learning mechanisms when NO can act as a volume transmitter within memory forming circuits. In particular, the molecular analysis of learning mechanisms (most notably in insects and gastropod molluscs) opens conceptually different perspectives to understand the logic of recruiting evolutionarily conserved pathways for novel functions. Giant uniquely identified cells from Aplysia and related species precent unuque opportunities for integrative analysis of NO signaling at the single cell level. PMID- 21622166 TI - Identification of the Lactobacillus SLP domain that binds gastric mucin. AB - Surface layer proteins (SLPs) of lactobacillus bacteria have some structural regions responsible for adhesion to the intestinal epithelium. To identify the SLP and the smallest domain within the protein that is responsible for the adhesion of the bacterium to the intestinal epithelium, L. plantarum strain CGMCC1258 was investigated in this study. Using bioinformatics and molecular techniques, we first identified and purified a novel protein, integral membrane protein-2 (IMP-2, 33-45 kDa) responsible for adhesion to gastric mucin. Truncated forms of IMP-2 were then constructed and expressed, and the amino acids from 515 to 575 (designated micro IMP, MIMP) was identified as the smallest domain responsible for adhesion to gastric mucin. Competing assay was performed, which further confirmed the ability of MIMP to compete with enteroinvasive E. coli and enteropathogenic E. coli to adhere to cells of a normal colon cell line, NCM460. Furthermore, MIMP could maturate dendritic cells. These findings set a foundation for further investigation on the role of MIMP in the treatment and prevention of inflammation-related diseases of the intestine. PMID- 21622167 TI - STIM and Orai in hemostasis and thrombosis. AB - At sites of vascular injury, platelets rapidly adhere to the exposed subendothelial extracellular matrix, become activated and, together with the coagulation system, form a plug that seals the lesion. This process is essential to prevent blood loss, however, under pathological conditions it may lead to vessel occlusion. Agonist-induced elevation of intracellular Ca(2+) levels is essential for platelet activation. It occurs through two different mechanisms: Ca(2+) release from internal stores, involving phospholipase C (PLC)-dependent generation of inositol-1,4,5-trisphosphate (IP3) and activation of IP3 sensitive channels in the store membrane, and Ca(2+) influx across the plasma membrane. Store operated Ca(2+) entry (SOCE), triggered by store depletion, is the main influx pathway for extracellular Ca(2+) in platelets, but the molecular mechanism underlying this pathway has long remained elusive. In the last years, however, the Ca(2+) sensor stromal interaction molecule 1 (STIM1) and the channel protein Orai1 emerged as the key players in platelet SOCE. This review summarizes the current knowledge about the role of these proteins in platelet physiology and thrombus formation and discusses their suitability as antithrombotic targets. PMID- 21622168 TI - The role of miRNAs in cytokine signaling. AB - This review explores the relationship between cytokines and microRNAs (miRNAs). In particular, the regulation of miRNAs by pro-inflammatory cytokines, anti inflammatory cytokines, interferons and transforming growth factor beta are examined, highlighting how miRNAs can mediate some of the known functions of these cytokines, as well as identifying novel gene targets, mechanisms and cross talk between pathways. PMID- 21622169 TI - Purinoceptors in inflammation: potential as anti-inflammatory therapeutic targets. AB - Purinergic receptors or purinoceptors are expressed in many mammalian cells and are activated by extracellular purines (adenine, purine nucleotides and nucleosides). Both adenosine (P1) and nucleotide/nucleoside (P2, grouped in P2X and P2Y subtypes) receptors exert important role in the inflammatory processes. The significative up-regulation of many purinoceptors located on the immune cells (neutrophils, eosinophils, monocytes, macrophages, mast cells and lymphocytes) in the course of inflammatory diseases supports the interpretation of their functions. New insights into the involvement of purinoceptors also in the neuro inflammatory diseases (e.g. conditions of chronic inflammation associated with neurodegenerative diseases) are proposed. The identification of antagonists of purinergic receptors potentially useful to control inflammatory pathways represents the object of many studies reported in the recent literature. Aim of this review is to recapitulate the most recent data and experimental findings that highlight the critical, double edge, effect of these receptors in inflammation, making consistent the possibility to target them to control and regulate inflammation. PMID- 21622170 TI - Microglial response to viral challenges: every silver lining comes with a cloud. AB - Microglia, the resident macrophages of the Central Nervous System (CNS) mediate key innate immune responses against foreign invasions within the CNS and clear the debris after any damage to the nearby tissue. Blood Brain Barrier (BBB) segregates the CNS from the rest of the lymphatic system and prevents the entry of foreign molecules into the brain. Pathogens still cross the BBB via different mechanisms and can cause severe infections of the CNS. Viral encephalitis is the most common form of brain infection and the causative agents include Japanese Encephalitis Virus (JEV), West Nile Virus (WNV), Murray Valley Encephalitis Virus (MVEV), Herpes Simplex Virus (HSV), Human Immunodeficiency Virus (HIV), Cytomegalovirus (CMV) and Hepatitis C Virus (HCV) among several others. Microglia expresses various Pattern Recognition Receptors (PRRs) to identify viral signatures called Pathogen Associated Molecular Patterns (PAMPs) to which microglia respond by releasing several pro and anti-inflammatory cytokines like MCP1, IL-1beta, Type I IFN, IFN-gamma, TNF-alpha etc. This review discusses the various viral infections of the brain and strategies employed by microglia to detect them. PMID- 21622171 TI - An extended Myc network contributes to glucose homeostasis in cancer and diabetes. AB - The Myc network of transcription factors plays pleiotropic roles in normal and pathological cell function. The canonical Myc network controls how the essential nutrients glucose and glutamine are utilized inside cells. The Myc network carries out this function by upregulating glucose and glutamine transporters and key enzymes in the glycolytic or glutaminolytic pathways. The Myc network also coordinates cellular utilization of glucose and glutamine in biosynthetic pathways by directly regulating mitochondrial mass and activity. We present an argument for the existence of an "extended" Myc network comprised of two related transcription factors MondoA and ChREBP. Both MondoA and ChREBP sense glycolytic flux and are the principal regulators of glucose-dependent transcription in their respective tissues, skeletal muscle and liver. MondoA also senses glutaminolytic flux into the tricarboxylic acid cycle and appears to coordinate the utilization of glucose and glutamine by regulating expression of thioredoxin interacting protein. Current data suggest that the extended Myc network regulates the cellular response to changes in nutrient availability and may be altered in cancer and insulin resistance. PMID- 21622172 TI - Targeting rapid action of sex steroid receptors in breast and prostate cancers. AB - Human breast and prostate cancers are complex diseases caused by the progressive accumulation of gene mutations combined with epigenetic deregulation of critical genes and derangement of signaling pathways. Compelling evidence indicates that steroid hormones elicit non-genomic responses in cytoplasm of target cells. In this cellular location, steroid-coupled receptors recruit signaling effectors or scaffold proteins, thereafter activating multiple pathways leading to proliferation, survival, migration and invasiveness. Thus, the immediate challenge is the dissection of key upstream events regulating steroid response in target tissues to prevent progression and improve treatment of breast and prostate cancers. Progress in our understanding of the molecular mechanisms that play a master role in these cancers has strongly stimulated the search for specific inhibitors of key signaling molecules. This review aims to give an up-to date report of the complex network regulating non-genomic action of steroid hormones in target cells. The final section highlights recent advances from our laboratory and future directions in alternative approaches for the treatment of breast and prostate cancers. PMID- 21622173 TI - Dramatic improvement of DC-based immunotherapy against various malignancies. AB - Dendritic cells (DCs) play a crucial role in maintaining the immune system. Though DC-based cancer immunotherapy has been suggested as a potential treatment for various kinds of malignancies, clinical efficacies are still insufficient in many human trials. To identify the causes of the low efficacies, we paid attention to their numbers and how they are activated. We proved that DCs' antitumor effect depends on their number and the way they are activated. We here established a possible breakthrough, a simple cytokine-based culture method to realize a log-scale order of functional murine/human DCs. Moreover, we demonstrated that DCs activated by replication-deficient recombinant Sendai virus (rSeV) were dramatically more effective than that seen in the use of current DC vaccine for immunotherapy against malignancies. Our study could overcome these problems and would improve treatment of malignancies. PMID- 21622174 TI - Animal models to study cancer-initiating cells from glioblastoma. AB - Three main subtypes of gliomas with distinct molecular pathologies have been modeled in animals to better understand their biology. Genetically engineered mouse models that take advantage of genetic abnormalities observed in human gliomas have been instrumental in this process. These models better recapitulate signaling transduction pathways and the microenvironment that play crucial roles in glioma formation than in vitro systems or transplantation models. An increasing amount of data supports the existence of cells functionally defined by their self-renewal ability and tumor-initiating potential upon serial transplantation. As the issue of these cells with stem cell character in gliomagenesis becomes more illusive, animal models that provide an accurate experimental system where the stem cell character can be manipulated and studied are urgently needed. This review provides an overview of the current state of the literature with respect to animal models used in the study of gliomas and cells with stem cell character in their native environment. PMID- 21622175 TI - RAS and Hedgehog--partners in crime. AB - Both RAS and Hedgehog (HH) pathway activation can be found in approximately one third of all cancers. In many cases, this activation occurs in the same tumor types, suggesting a positive impact of a simultaneous activation of RAS and HH on tumor development. This review aims to summarize the current knowledge about the molecular and functional crosstalk of RAS and HH signaling in the development of hyperproliferative disease. PMID- 21622176 TI - Mesenchymal stem cells and their microenvironment. AB - Mesenchymal stem cells (MSC) are multipotent stem cells that hold promise for an expanding list of therapeutic uses, not only due to their ability to differentiate into all connective tissues including bone, fat and cartilage, but additionally due to their trophic and anti-inflammatory effects which contribute to healing and tissue regeneration. Ongoing research is starting to illuminate important aspects of the microenvironmental niche, which supports MSC self renewal. In this review, we summarize recent findings on cellular structures and molecular pathways that are involved in regulation of MSC self-renewal versus differentiation, and in retention of MSCs within the niche versus mobilization and recruitment to sites of injury. In addition, the contribution of MSCs to the structure and function of hematopoietic and cancerous niches is discussed. PMID- 21622177 TI - Protein-ligand docking. AB - Ligand-docking is an established computational technique universally applied in structure-based drug design. Since the first attempts carried out in the early '80s to predict the three-dimensional conformation of a protein-ligand bound complex, this methodology has evolved constantly and it is presently implemented in many different ways. The present study aims at explaining the standard protein ligand docking protocol, together with its main advantages and drawbacks. Milestone reports and future directions are reported and discussed as well. PMID- 21622179 TI - Role of purinergic signalling in neuro-immune cells and adult neural progenitors. AB - Inflammation has a key role in a vast range of central nervous system diseases. Under acute and chronic neurodegenerative conditions, resident microglia and astrocytes and blood-borne immune cells concur to neuroinflammation and remodeling/repair at the inflammed site. Distinct inflammatory cell states characterized by either a beneficial or a detrimental phenotype have been identified, depending upon the timing after the initial insult and activation by specific pro- or anti-inflammatory molecules. Of note, quiescent adult neuroprogenitor cells located in both brain's neurogenic areas and parenchyma have been recently shown to interact with immune cells and actively participate to restore function. Among other systems, extracellular nucleotides and their receptors have emerged as early alerting signals in inflammation and as key players in orchestrating the release of inflammatory molecules and the interaction between different cell types. Here, we revise the state of the art in this expanding field with the final aim of unveiling whether new purinergic-based therapies may be useful to halt excessive inflammation and foster endogenous repair in neuroinflammatory disorders. PMID- 21622178 TI - Modulation of neoplastic gene regulatory pathways by the RNA-binding factor AUF1. AB - The mRNA-binding protein AUF1 regulates the expression of many key players in cancer including proto-oncogenes, regulators of apoptosis and the cell cycle, and pro-inflammatory cytokines, principally by directing the decay kinetics of their encoded mRNAs. Most studies support an mRNA-destabilizing role for AUF1, although other findings suggest additional functions for this factor. In this review, we explore how changes in AUF1 isoform distribution, subcellular localization, and post-translational protein modifications can influence the metabolism of targeted mRNAs. However, several lines of evidence also support a role for AUF1 in the initiation and/or development of cancer. Many AUF1-targeted transcripts encode products that control pro- and anti-oncogenic processes. Also, overexpression of AUF1 enhances tumorigenesis in murine models, and AUF1 levels are enhanced in some tumors. Finally, signaling cascades that modulate AUF1 function are deregulated in some cancerous tissues. Together, these features suggest that AUF1 may play a prominent role in regulating the expression of many genes that can contribute to tumorigenic phenotypes, and that this post-transcriptional regulatory control point may be subverted by diverse mechanisms in neoplasia. PMID- 21622180 TI - Pulmono-atrial shunt and lung assist to treat right ventricular failure. AB - In right ventricular failure (RVF) a decrease of right ventricular afterload and improvement of left atrial filling could be achieved by a pulmonary artery-left atrial (PA)-shunt. To avoid cyanosis, artificial oxygenation is necessary. In 11 pigs a PA-shunt was created. An interventional lung assist device (ILA) was installed from the femoral artery to vein in 5 pigs (serial in relation to native lung: Group I) and into the PA-shunt in 6 pigs (parallel: Group II). RVF was induced by pulmonary artery banding. Right ventricular performance was determined by pulse contour analysis, pressure - and flow measurements. In both groups a stable RVF was generated. In Group I cardiac output trended to increase but neither right ventricular filling pressures nor arterial pressure changed significantly. The PaO2 decreased significantly. In Group II cardiac output and arterial pressure increased significantly under a shunt flow of 2.3- 2.6 l/min and the animals recovered from cardiogenic shock. In conclusion a PA-shunt with a parallel lung assist can effectively reverse the deleterious effects of RVF. PMID- 21622181 TI - The role of Wnt in cell signaling and cell adhesion during early vertebrate development. AB - During embryonic development, a group of dividing blastomeres is ultimately shaped into a structured, functional organism. To achieve this goal, individual cells and groups of cells need to move to new positions, organize themselves, and differentiate into specialized cell types. In these processes, intercellular contacts and contacts between cells and their environment play critical roles. The cells interact physically via cell adhesion molecule and communicate through signaling pathways. One of the pathways active during embryonic development is the Wnt pathway. Interestingly, Wnt and cell adhesion are often active in the same processes and crosstalk between them exists by reciprocal regulation and sharing of components. In this review, we will focus on how Wnt signaling cooperates with cell adhesion to ensure smooth processing of gastrulation, somitogenesis and neurulation. PMID- 21622182 TI - Cerebral arterioles preparation and PECAM-1 expression in C57BL/6J and ApoE-/- mice. AB - Exploration of molecular mechanisms involved in alterations of cerebral blood vessels in pathology such as chronic kidney disease implies cerebral vessel isolation for proteins and mRNAs quantification. We describe an updated method to isolate brain microvessels from wild type (WT) mice at 14 weeks of age. The quality of cerebral arterioles preparation was determined by measuring the enzymatic activity of gamma-glutamyl transpeptidase, an enzyme that is especially expressed in cerebral microvessels, and the quantitative expression (Western Blot) of platelet endothelial cell adhesion molecule-1 (PECAM-1), alpha-actin and gamma-enolase as markers of respectively endothelial cells, smooth muscle cells and neurons. We then assessed PECAM-1 expression in 14 and 20 weeks old ApoE(-/-) and WT mice. PECAM-1 expression was increased in 14 weeks old ApoE(-/-) mice compared to age matched WT mice. This difference in PECAM-1 expression disappeared at 20 weeks of age. These findings indicate that the present method of mice cerebral arterioles isolation is a valid method. Moreover, PECAM-1 expression, a marker of endothelial cells, changes with age in cerebral arterioles of ApoE(-/-) mice. PMID- 21622183 TI - Protective role of heme oxygenase-1 against inflammation in atherosclerosis. AB - Heme oxygenase-1 (HO-1) catalyzes the first and rate-limiting step in the metabolism of free heme into equimolar amounts of ferrous iron, carbon monoxide (CO), and biliverdin. Biliverdin is subsequently converted to bilirubin by biliverdin reductase. HO-1 has recently been identified as a promising therapeutic target in the treatment of vascular inflammatory disease, including atherosclerosis. HO-1 represses inflammation by removing the pro-inflammatory molecule heme and by generating CO and the bile pigments, biliverdin and bilirubin. These HO-1 reaction products are capable of blocking innate and adaptive immune responses by modifying the activation, differentiation, maturation, and/or polarization of numerous immune cells, including endothelial cells, monocytes/macrophages, dendritic cells, T lymphocytes, mast cells, and platelets. These cellular actions by CO and bile pigments result in diminished leukocyte recruitment and infiltration, and pro-inflammatory mediator production within atherosclerotic lesions. This review highlights the mechanisms by which HO 1 suppresses vascular inflammation in atherosclerosis, and explores possible therapeutic modalities by which HO-1 and its reaction products can be employed to ameliorate vascular inflammatory disease. PMID- 21622184 TI - MicroRNA-regulated transgene expression systems for gene therapy and virotherapy. AB - For safe and effective gene therapy, targeted tissue-restricted transgene expression is desirable. Various methods have been developed to achieve such expression, including the use of tissue-specific promoters. In addition to these approaches, a new system which can regulate transgene expression, including viral gene expression, by exploiting microRNAs (miRNAs) has recently been developed. miRNAs are approximately 22-nucleotide (nt)-long non-coding RNAs that translationally suppress or catalytically degrade target mRNA through binding to imperfectly complementary sequences in the 3'-untranslated region (UTR). In miRNA regulated transgene expression systems, tandem copies of sequences perfectly complementary to the miRNAs are usually incorporated into the 3'-UTR of the transgene expression cassette, leading to the suppression of transgene expression in cells expressing the corresponding miRNAs. miRNA-mediated regulation of transgene expression was first demonstrated for lentivirus vectors, and subsequently this technology was applied to replication-incompetent adenovirus vectors, tumor-specific oncolytic viruses for cancer therapy, and recombinant live attenuated viruses for vaccine therapy. The aim of this review is to highlight the applications of miRNA-regulated transgene expression systems for gene therapy and virotherapy. PMID- 21622186 TI - Ingression, migration and early differentiation of cardiac progenitors. AB - During vertebrate embryogenesis the heart is the first functioning organ and cardiac progenitor cells (CPCs), which form the future heart, are among the first cell types to be established during gastrulation. A large number of studies indicate that cardiac development is tightly regulated by a series of molecular signaling pathways and morphological events. The cellular and molecular events that control early cardiac development are conserved among vertebrates. The favorable experimental characteristics of the chicken embryo and the ease in which cell labeling and imaging can be performed has allowed direct observation of the process of gastrulation and cell migration trajectories. This has enabled the study of the signaling proteins and molecular pathways required to specify early embryonic cells to the myocardial lineage. In this review we discuss the major morphogenetic and regulatory events that control gastrulation and migration of CPCs in the chicken embryo. We also describe the signaling mechanisms critical for early CPC specification in pre-gastrula, gastrula and early neurula stage embryos. PMID- 21622185 TI - WT1/EGR1-mediated control of STIM1 expression and function in cancer cells. AB - There have been numerous publications linking Ca(2+) signaling and cancer, however, a clear explanation for this link has remained elusive. We recently identified the oncogenes/tumor suppressors Wilms Tumor Suppressor 1 (WT1) and Early Growth Response 1 (EGR1) as regulators of the expression of STIM1, an essential regulator of Ca(2+) entry in non-excitable cells. The current review focuses on the literature defining both differential Ca(2+) signaling and WT1/EGR1 expression patterns in 6 specific cancer subtypes: Acute Myeloid Leukemia, Wilms Tumor, breast cancer, ovarian cancer, glioblastoma and prostate cancer. For each tumor-type, we have assessed how specific changes in WT1 and EGR1 expression might contribute to aberrant Ca(2+) homeostasis as well as the therapeutic potential of these observations. PMID- 21622187 TI - An update on the role of carboxypeptidase U (TAFIa) in fibrinolysis. AB - Since its discovery more than 20 years ago, a lot has been revealed about the biochemistry and physiological behaviour of carboxypeptidase U (CPU). Recent advances in CPU research include the unravelling of the crystal structure of proCPU and revealing the molecular mechanisms for the marked instability of the active enzyme, CPU. The recent development of two highly sensitive assays has cleared the path toward the direct measurement of CPU in circulation or the determination of CPU generation, rather than the measurement of total proCPU concentration in plasma. Finally, since CPU is known to have a prominent bridging function between coagulation and fibrinolysis, the development of CPU inhibitors as profibrinolytic agents is an attractive new concept and has gained a lot of interest from several research groups and from the pharmaceutical industry. These recent advances in CPU research are reviewed in this literature update. PMID- 21622188 TI - Metabonomics and its role in amino acid nutrition research. AB - Metabonomics combines metabolic profiling and multivariate data analysis to facilitate the high-throughput analysis of metabolites in biological samples. This technique has been developed as a powerful analytical tool and hence has found successful widespread applications in many areas of bioscience. Metabonomics has also become an important part of systems biology. As a sensitive and powerful method, metabonomics can quantitatively measure subtle dynamic perturbations of metabolic pathways in organisms due to changes in pathophysiological, nutritional, and epigenetic states. Therefore, metabonomics holds great promise to enhance our understanding of the complex relationship between amino acids and metabolism to define the roles for dietary amino acids in maintaining health and the development of disease. Such a technique also aids in the studies of functions, metabolic regulation, safety, and individualized requirements of amino acids. Here, we highlight the common workflow of metabonomics and some of the applications to amino acid nutrition research to illustrate the great potential of this exciting new frontier in bioscience. PMID- 21622189 TI - Structure and function of the thymic microenvironment. AB - Organs are more than the sum of their component parts--functional competence requires that these parts not only be present in the appropriate proportions, but also be arranged and function together in specific ways. The thymus is an excellent example of the connection between cellular organization and organ function. Unlike more familiar organs, such as lung or kidney, the thymus is not organized into easily identifiable structures such as tubes and ordered cell layers, but instead is a complex meshwork of microenvironments through which T cell progenitors migrate, receiving signals that instruct them to differentiate, proliferate, or die. Proper thymic organization is essential to the optimal production of a functional T cell repertoire. During aging, the thymus undergoes involution, largely due to degradation of the TEC microenvironmental compartment, which then fails to support optimal thymocyte development resulting in reduced output of naive T cells. This review will summarize the current state of understanding of the composition and organization of thymic microenvironments and the mechanisms that promote their proper development and function. PMID- 21622190 TI - Environmental endocrine disruptors: does a sex-related susceptibility exist? AB - Several substances present in the environment, now classified as endocrine disruptors (EDs), strongly interfere with both androgen and oestrogen actions in reproductive tissues. However, nowadays it is well recognized that these sex steroid hormones are more than regulators of gonadal functions. In fact, they, in synergy with genes, are responsible of sex-related differences in anatomical, physiological, and behavioral traits which characterize males and females of many vertebrate species, including humans. Thus, even if EDs are present in minute amount (part for trillion) in environment, their effects in male and female physiology could be greater than before expected also prejudicing the sex-steroid hormone-induced integrated physiological responses in women and men. In addition, differences in male and female susceptibility to EDs could be present even if scarce information on this aspect is still available. Here we have reviewed the state of the art on the sex-related susceptibility to EDs underlying the mechanism at the root of these effects. PMID- 21622191 TI - Significance of membrane microparticles in solid graft and cellular transplantation. AB - Microparticles (MPs) are submicron vesicles released from stimulated or apoptotic cells after plasma membrane remodeling. In body fluids, they constitute relevant hallmarks of cell damage. Having long been considered inert debris reflecting cellular activation or damage, MPs are now considered as cellular effectors involved in cell-cell crosstalk. This review focuses on the pathophysiologic significance of MPs in the particular setting of solid graft and cellular transplantation. PMID- 21622192 TI - Cytokines as prognstic tool in breast carcinoma. AB - Serum cytokines are promising biomarkers of cancer staging and outcome prediction, including response to treatment. Serum samples were collected from 200 breast carcinoma patients prior to chemotherapy treatment. Luminex liquid protein chip technology was used to analyze 25 cytokines in serum. Linear regression was used to analyze the relationship of cytokine levels and tumor size. The independent sample T-test and Chi-square test methods were used to analyze the difference of cytokine levels between two groups. IL-12p40, sIL-2R, MMP-2 levels showed linear correlation with tumor size. Eotaxin, IL-10, IL-12p70, IL-7, IL-1ra, IP-10, MCP-1beta, MP-2 and MIP-1beta levels showed significant difference between different lymph node groups, but only Eotaxin, IP-10 and MCP-1 levels had an inverse correlation with the number of positive nodes. Fractalkine, G-CSF, MIP-1alpha, MIP-1beta levels showed significant differences between different ER+ groups. Eotaxin, Fractalkine, IL-6, IL-7, IL-10, MCP-1 and VEGF levels had significant differences between different HER-2 groups. Our study resulted in the identification of a serum cytokine profile with the potential to be clinically applicable to predict disease outcome and in monitoring of efficacy of treatment. PMID- 21622193 TI - Experimental study of 99mTc-depreotide preparation and its affinity with A549 cell. AB - The (99m)Tc-labeled agent, ((99m)TcO)depreotide, has received regulatory approval in the United States and Europe for use in the detection of cancer. It is essential to establish a simple and reliable method of direct radiolabeling of (99m)Tc-depreotide and to investigate its specific receptor binding properties with human non small cell lung cancer (NSCLC) A549 cell in vitro. So we made some researches as follow: Depreotide was labeled with (99m)Tc using SnCl2 as a reductant. Labeling efficiencies at different pH values and temperatures were compared. Radioreceptor assay was used to observe the uptake kinetics, stagnation and retention half time of (99m)Tc-depreotide in A549 cells. As the results of the investigation ,many facts is shown below: The labeling rate of pH 6.0 group was higher than that of pH 5.0 and pH7.0 groups. The labeling rate decreased when temperature increased from 15 degrees C to 50 degrees C. The uptake rate increased with rising temperature, and the maximum uptake was observed at 60 min at 37 degrees C. The cleaning curves were similar at different temperatures, and the half cleaning time at 37 degrees C was 48 min. The results showed that the optimal conditions for labeling depreotide with (99m)Tc was found to be below 15 degrees C at a pH lower than 6.0. Furthermore, at 37 degrees C, (99m)Tc depreotid may have the potential as an ideal imaging agent for somatostatin receptors. PMID- 21622194 TI - Organic matrix-related mineralization of sea urchin spicules, spines, test and teeth. AB - The camarodont echinoderms have five distinct mineralized skeletal elements: embryonic spicules, mature test, spines, lantern stereom and teeth. The spicules are transient structural elements whereas the spines, and test plates are permanent. The teeth grow continuously. The mineral is a high magnesium calcite, but the magnesium content is different in each type of skeletal element, varying from 5 to 40 mole% Mg. The organic matrix creates the spaces and environments for crystal initiation and growth. The detailed mechanisms of crystal regulation are not known, but acidic and phosphorylated matrix proteins may be of special importance. Biochemical studies, sequencing of the complete genome, and high throughput proteomic analysis have not yet provided insight into the mechanisms of crystallization, calcite composition, and orientation applicable to all skeletal elements. The embryonic spicules are not representative of the mature skeletal elements. The next phase of research will have to focus on the specific localization of the proteins and individual biochemistries of each system with regard to mineral content and placement. PMID- 21622195 TI - Role of Rho GTPases and their regulators in cancer progression. AB - Rho family of GTPases is an ubiquitiously expressed and evolutionarily conserved family of GTP binding proteins that regulate actin dynamics and intracellular signaling. Among the Rho family GTPases, three members RhoA, Rac1 and CDC42 have been well characterized. They each play pivotal roles in gene expression, cell proliferation, apoptosis and various cellular functions. They are driven by signaling from RhoGDIs, RhoGEFs, RhoGAPs and cell surface receptors. Abnormalities in Rho GTPase function have major consequences on cell behavior. Over expression of Rho GTPases is associated with reorganization of actin cytoskeleton, an increase in cell migration, invasion and metastasis which are important aspects of cancer progression. This review will explore these Rho GTPases and the function of their associated signaling pathways in different types of cancers. PMID- 21622196 TI - Role of purinergic receptor polymorphisms in human bone. AB - Osteoporosis is a multifactorial disease with a strong genetic component. Variations in a number of genes have been shown to associate with bone turnover and risk of osteoporosis. P2 purinergic receptors are proteins that have ATP or other nucleotides as their natural ligands. Various P2Y and P2X receptor subtypes have been identified on bone cells. Several cellular functions in bone tissue are coupled to P2-receptor activation, including bone resorption, cytokine release, apoptosis, bone formation, and mineral deposition. Furthermore, ATP release and P2 purinergic signalling is a key pathway in the mechanotransductory process, where mechanical stimulation on bone leads to anabolic responses in the skeleton. A number of single nucleotide polymorphisms have been identified in the P2 receptor genes, where especially the P2X7 subtype has been the focus of extensive investigation where several polymorphisms have been shown to have functional implications on receptor function; moreover, some polymorphisms are associated with alterations in bone turnover and bone mass. This review focuses on variations in P2 receptor genes and the association to bone turnover and quality. PMID- 21622197 TI - Genomic instability caused by hepatitis B virus: into the hepatoma inferno. AB - Chronic hepatitis B virus (HBV) infection is an important cause of hepatocellular carcinoma (HCC) worldwide, especially in Asia. HBV induces HCC through multiple oncogenic pathways. Hepatitis-induced hepatocyte inflammation and regeneration stimulates cell proliferation. The interplay between the viral and host factors activates oncogenic signaling pathways and triggers cell transformation. In this review, we summarize previous studies, which reported that HBV induces host genomic instability and that HBV-induced genomic instability is a significant factor that accelerates carcinogenesis. The various types of genomic changes in HBV-induced HCC--chromosomal instability, telomere attrition, and gene-level mutations--are reviewed. In addition, the two viral factors, HBx and the pre-S2 mutant large surface antigen, are discussed for their roles in promoting genomic instability as their main features as viral oncoproteins. PMID- 21622199 TI - Applications of proteomics in cartilage biology and osteoarthritis research. AB - In osteoarthritis (OA) the turnover of extracellular matrix (ECM) macromolecules is disrupted by catabolic changes that lead to the production of a range of inflammatory mediators and the loss and fragmentation of proteoglycans, fibrillar and non-fibrillar collagens. These events result in the degradation and release of ECM fragments, which are potential biomarkers that can be detected in synovial fluid, blood and urine. Proteomics is increasingly applied in cartilage research and has the potential to advance our understanding of the biology of this tissue. It can also provide mechanistic insight into disease pathogenesis and progression and facilitate biomarker discovery. Here we review the area of cartilage and chondrocyte proteomics and published studies relevant to arthritis and OA biomarkers, highlighting areas of current and future research and development. Markers of tissue turnover in joints have the capacity to reflect disease relevant biological activity potentially enabling a more rational approach to healthcare management. Therefore proteomic studies of cartilage, chondrocytes and their subcellular fractions and other joint cells and tissues may be particularly relevant in diagnostic orthopedics and therapeutic research. PMID- 21622200 TI - Hepatocyte aquaporins in bile formation and cholestasis. AB - Bile formation by hepatocytes is an osmotic secretory process that is ultimately dependent on the biliary secretion of osmotically-active solutes (mainly bile salts) via specialized canalicular transporters as well as on the water permeability of the canalicular plasma membrane domain. Hepatocytes express aquaporins, a family of membrane channel proteins that facilitate the osmotically driven movement of water molecules. Aquaporin-8 (AQP8), localized to canalicular membranes, modulates membrane water permeability providing a molecular mechanism for the osmotically-coupled transport of solute and water during bile formation. There is experimental evidence suggesting that defective hepatocyte AQP8 expression leads to alterations in normal bile physiology. Thus, AQP8 protein is downregulated (and canalicular water permeability decreased), in established rat models of cholestasis, such as sepsis-associated cholestasis, estrogen-induced cholestasis and extrahepatic obstructive cholestasis. Moreover, AQP8 gene silencing in the human hepatocyte-derived cell line HepG2 inhibits canalicular water secretion. Based on current knowledge, it is conceivable that cholestasis results from a mutual occurrence of impaired solute transport and AQP8-mediated decrease of canalicular water permeability. PMID- 21622201 TI - Omega-3 polyunsaturated fatty acids in the brain: metabolism and neuroprotection. AB - Omega-3 polyunsaturated fatty acids (n-3 PUFAs) are a group of essential fatty acids that serve as energy substrates and integral membrane components, and therefore play crucial roles in the maintenance of normal neurological function. Recent studies show that n-3 PUFAs display neuroprotective properties and exert beneficial effects on the cognitive function with aging. The brain's need of n-3 PUFAs is predominantly met by the blood delivery due to their limited synthesis in the brain. The present review focuses on the metabolism of n-3 PUFAs in the brain, including their accumulation and turnover. We also highlight the current understanding of the neuroprotective effects of n-3 PUFAs against cerebral ischemia and neurodegenerative disorders, such as Alzheimer's disease and Parkinson's disease. PMID- 21622198 TI - Biomineralization of bone: a fresh view of the roles of non-collagenous proteins. AB - The impact of genetics has dramatically affected our understanding of the functions of non-collagenous proteins. Specifically, mutations and knockouts have defined their cellular spectrum of actions. However, the biochemical mechanisms mediated by non-collagenous proteins in biomineralization remain elusive. It is likely that this understanding will require more focused functional testing at the protein, cell, and tissue level. Although initially viewed as rather redundant and static acidic calcium binding proteins, it is now clear that non collagenous proteins in mineralizing tissues represent diverse entities capable of forming multiple protein-protein interactions which act in positive and negative ways to regulate the process of bone mineralization. Several new examples from the author's laboratory are provided which illustrate this theme including an apparent activating effect of hydroxyapatite crystals on metalloproteinases. This review emphasizes the view that secreted non-collagenous proteins in mineralizing bone actively participate in the mineralization process and ultimately control where and how much mineral crystal is deposited, as well as determining the quality and biomechanical properties of the mineralized matrix produced. PMID- 21622202 TI - Serum biomarker of diabetic peripheral neuropathy indentified by differential proteomics. AB - At least one in four diabetic patients is affected by peripheral neuropathy. In this study, the MALDI-TOF-MS mass spectra of peptides and proteins were generated following WCX CLINPROT bead fractionation of 39 diabetic peripheral neuropathy (DPN), 39 diabetes mellitus (DM), and 35 control (CON) serum samples. The spectra were analyzed statistically using flexAnalysisTM and Clin-ProtTM bioinformatics software. Identification of the selected markers was performed and affinity bead purified plasma protein was subjected to LTQ Orbitrap XL MS/MS analysis followed by Mascot identification of the peptide sequences. 89 differentially expressed peaks of serum proteins were identified. 17, 10 and 4 most significant peaks between CON vs. DM, CON vs. DPN, DM vs. DPN, respectively, were selected out using the ClinProTool software package and used to train a Supervised Neural Network. A veracity rate of 100% was obtained for all sets. Following this analysis, a 6631-Da marker was identified as a fragment of the Apolipoprotein C-I precursor. The peptides identified may have clinical utility as surrogate markers for detection and classification of DM and DPN. PMID- 21622203 TI - Epigenetic regulation in cancer development. AB - From an operational definition of epigenetic, we move to provide the reader a general but comprehensive description of epigenetic phenomena that often lead to cell transformation. The last decade has, in fact, seen novel players involved in the regulation of gene expression. Not only protein factors but also a number of chromatin modifiers and remodelling proteins, which regulate the level of compaction of the genome through a variety of post-translational modifications deposed on histone tails or on DNA itself. Meanwhile, the discovery of tiny RNAs, of only 21-23 nucleotides in length, has brought to the attention their role as key regulators in the cell, being able to direct differentiation programs and function as oncogenes or oncosuppressors. In this general compendium, we aim to describe main cellular functions that through an epigenetic or epigenetic associated mechanism have been found to be directly implicated in cancerogenesis. PMID- 21622204 TI - Neuroglialpharmacology: white matter pathophysiologies and psychiatric treatments. AB - Psychotropic treatments such as second generation or atypical antipsychotics are efficacious in a wide spectrum of psychiatric disorders ranging from schizophrenia to depression, bipolar disorder, and autism. These treatments are associated with peripheral metabolic derangements that are often also present in drug-naive patients. Furthermore, altering lipid composition/levels (with omega 3 fatty acids) and ameliorating oxidative toxicities may treat/prevent disease. The above observations are reexamined from the perspective of a myelin-centered model of the human brain. The model proposes that the human brain's extensive myelination required higher metabolic resources that caused evolutionary adaptations resulting in our quadratic (inverted U) myelination trajectory that peaks in the sixth decade of life. It further proposes that optimal brain function depends on exquisite action potential synchronization that myelin makes possible and that myelin's exceptional vulnerability to subtle metabolic/oxidative abnormalities may promote both developmental and degenerative diseases. Available data are integrated herein to suggest that widely used psychotropic treatments have under-appreciated CNS metabolic and neurotransmitter effects on myelination, its plasticity, and repair that may substantially contribute to their mechanisms of action. PMID- 21622205 TI - Induction and patterning of intramembranous bone. AB - The primary focus of this article is to review intramembranous bone development, that is, ossification that takes place directly. Comparisons with endochondral ossification (ossification with a cartilage precursor) will be made in order to illustrate the differences between these two modes of ossification and to highlight the comparatively sparse information that is available about intramembranous ossification. Despite decades of research into understanding skeletal development, there is still much to learn. Most of the research in this area has focused on the development of the calvariae (or skull bones) as typical intramembranous bones and the development of the limb bones as a typical endochondral bones. Few studies investigate other skeletal elements or compare these processes in a systematic manner. In this review, I focus primarily on condensation formation and skeletal patterning with specific examples from different organisms. PMID- 21622206 TI - The effect of dexamethasone and hypoxic stress on MC3T3-E1 cells. AB - Osteonecrosis of the femoral head (ONFH) can be caused by a decrease in the activity or numbers of osteoblasts, a process in which apoptosis may play an essential role. We investigated the effect of dexamethasone (Dex) combined with hypoxic stress on murine osteoblastic MC3T3-E1 cells. Flow cytometry, western blot and real-time quantitative PCR analyses revealed that hypoxia significantly enhanced Dex-induced apoptosis. Further data demonstrated that both the death receptor and the mitochondria-mediated pathway were involved in Dex-induced apoptosis under hypoxic conditions. However, the death receptor pathway had only a minor effect on this process. The expression levels of Bcl-2 and Bax, which regulate the mitochondria-initiated apoptotic cascade signaling pathway, were significantly different in response to Dex and hypoxia. The mitochondrial membrane potential collapsed, and the inhibitor brain- derived neurotrophic factor (BDNF) conferred effective protection against apoptosis. In summary, the mitochondria-mediated apoptotic pathway functions in osteoblast apoptosis that is induced by Dex in a hypoxic environment, and the present study may help us to gain further insight into the molecular mechanisms of steroid-induced ONFH. PMID- 21622207 TI - Alternative splicing of RAGE: roles in biology and disease. AB - The Receptor for Advanced Glycation End-products (RAGE) is a complex, multi ligand signaling system implicated in the pathogenesis of diabetes, cardiovascular disease and various cancers. RAGE undergoes extensive alternative splicing to produce a variety of transcripts with diverse functions, including soluble antagonists and variants with altered ligand binding domains. Studies focused on the major soluble variant (RAGEv1/esRAGE) have revealed this to function by binding RAGE-ligands and preventing activation of RAGE signaling in vascular and tumor cells. Furthermore, measurement of this variant in human serum has revealed that RAGEv1/esRAGE levels may represent a novel biomarker for RAGE ligand related pathogenic states. Understanding the full plethora of RAGE alternative splicing and its regulation is central to elucidating the role of RAGE in biology and disease. PMID- 21622208 TI - Leptin receptor signaling: pathways to leptin resistance. AB - The identification of spontaneous mutations in the leptin- and leptin receptor (ObR)-encoding ob and db gene, respectively, opened up a new field in obesity research. Leptin, an adipocyte-derived hormone, mirrors the body's fat stores and thereby informs the brain about the body's energy status. In the hypothalamus, leptin triggers specific neuronal subpopulations, like POMC and AgRP/NPY neurons, and activates several intracellular signaling events, including the JAK/STAT, MAPK, PI3K and mTOR pathway, which eventually translates into decreased food intake and increased energy expenditure. Leptin is also involved in the regulation of other physiological processes including reproduction, bone homeostasis and immune function. Here, we review the pathways that are activated upon ObR activation, how ObR expression is controlled and the molecular mechanisms leading to leptin resistance, i.e. the inability to adequately respond to elevated leptin levels and therefore a primary risk factor for obesity. PMID- 21622209 TI - JAK-STAT signaling in hepatic fibrosis. AB - Chronic liver injury, liver fibrosis and formation of hepatocellular carcinoma are intimately linked and represent a major medical challenge since treatment options are limited. Therefore, it is important to identify cellular and molecular pathways that promote liver damage or provide hepatoprotection for development of therapeutic approaches. Recently, the transcription factors STAT3 and STAT5 have been implicated in liver fibrosis induced by cholestatic liver damage. In this review, we summarize our current knowledge about STAT proteins in liver fibrosis and focus on common activities that underlie the hepatoprotective mechanisms regulated by IL-6/gp130/STAT3 and GH/STAT5/IGF-1 signaling pathways. PMID- 21622210 TI - Matrix vesicles: structure, composition, formation and function in calcification. AB - Matrix vesicles (MVs) induce calcification during endochondral bone formation. Experimental methods for structural, compositional, and functional analysis of MVs are reviewed. MV proteins, enzymes, receptors, transporters, regulators, lipids and electrolytes are detailed. MV formation is considered from both structural and biochemical perspectives. Confocal imaging of Ca(2+) and H(+) were used to depict how living chondrocytes form MVs. Biochemical studies revealed that coordinated mitochondrial Ca(2+) and Pi metabolism produce MVs containing a nucleational complex (NC) of amorphous calcium phosphate, phosphatidylserine and annexin A5--all critical to the mechanism of mineral nucleation. Reconstitution of the NC and modeling with unilamellar vesicles reveal how the NC transforms into octacalcium phosphate, regulated by Mg(2+), Zn(2+) and annexin A5. Extravasation of intravesicular mineral is mediated by phospholipases and tissue nonspecific alkaline phosphatase (TNAP). In the extravesicular matrix, hydroxyapatite crystal propagation is enhanced by cartilage collagens and TNAP, which destroys inhibitory PPi, and by metalloproteases that degrade proteoglycans. Other proteins also modulate mineral formation. Recent findings from single and multiple gene knockouts of TNAP, NPP1, ANK, PHOSPHO1, and Annexin A5 are reviewed. PMID- 21622211 TI - Molecular regulation of the intestinal epithelial barrier: implication in human diseases. AB - Intestinal barrier dysfunction is implicated in the development of various clinical diseases. While the study of intestinal barrier function has traditionally emphasized the impact of intestinal microflora and bacteria, the rapid development of molecular and cellular techniques has helped the recent transition of the field to the molecular regulation of the intestinal epithelial barrier. In this review, we summarized several aspects of recent progress on the molecular regulation of the intestinal epithelial barrier, ranging from the extrinsic factors such as probiotics, intrinsic protein effectors including the tight junction proteins, intestinal alkaline phosphatase and protein phosphatase 2A, to intestinal cell subsets such as intestinal intraepithelial lymphocytes and intestinal stem cells. Further investigations into the detailed mechanisms underlying the molecular regulation of the intestinal epithelial barrier will enable our manipulation of the factors and cell subsets involved to develop effective approaches to treat intestinal barrier dysfunction associated diseases. PMID- 21622212 TI - Crosstalk between viruses and PML nuclear bodies: a network-based approach. AB - Due to the recent advances in instrumental and scientific methods, cell biology data are generated with increasing speed and quantity. One of these fast developing fields is the crosstalk between promyelocytic leukemia protein nuclear bodies (PML-NBs) and viruses. PML-NBs are dynamic nuclear protein aggregates which are targeted by entire viral particles, viral proteins or viral nucleic acids. Their possible anti-viral properties motivated researchers to investigate the interaction between PML-NBs and viruses in depth. Based on extensive literature data mining, we created a comprehensive PML-NB/virus crosstalk Cytoscape network, which groups not only the most common relations but also less well described findings. The network is easy to navigate and provides a biologically relevant overview which can help finding interesting case studies. PMID- 21622213 TI - Complement in neurobiology. AB - The complement (C) system is a vital arm of innate immunity with many roles, including control of inflammation. This article examines the (C) system with emphasis on recent developments on complement relevant to neurobiology, in particular regarding our understanding and treatment of immune-mediated diseases. We will briefly outline the C system, and provide an updated review of its many receptors and regulatory factors. This section concludes with a listing of important roles of the C system, from recruitment of neural stem/progenitor cells, to its' relation to coagulation and adaptive immunity, and its lesser known but beneficial roles in physiology. We also review evidence for C-mediated diseases, which include multiple sclerosis and Alzheimer's disease. Therapeutic approaches for C-mediated diseases, considers emphasizing modulators of the C system including several less widely studied approaches such as heparinoids, vitamin D, and intravenous IgM. Finally, we summarize cutting-edge work on the role of C-mediated natural antibodies in autoimmunity and treatment strategies based on those findings, e.g., for remyelination and post-ischemic stroke repair. Improved understanding of the C system may hold great promise for the treatment of neurodegenerative diseases. PMID- 21622214 TI - Detection of EGFR and K-ras mutations for diagnosis of multiple lung adenocarcinomas. AB - The incidence of multiple primary lung adenocarcinoma (MPLA) is increasing, and it is important to distinguish MPLA from intrapulmonary metastasis (IPM) in order to determine the therapeutic strategy. However, there is no reliable method to differentiate between the two. The purpose of this study was to distinguish MPLA from IPM based on the gene status of EGFR and K-ras and the morphological Noguchi classification system. Sixty-eight tumors from 34 cases of clinical MPLA were evaluated. Of them, 11 cases (32.4%) were diagnosed as biological MPLA (bMPLA) by EGFR/K-ras mutation analyses, and 12 cases (35.3%) by morphological analysis. In all, 23 of the 34 cases (67.6%) were diagnosed as bMPLA. The remaining 11 cases were diagnosed as biological IPM (bIPM). The 5-year survival rates of bMPLA and bIPM were 90.9% and 63.6%, respectively (p=0.04). These findings suggest that the combination method including gene mutation and morphological analysis can guide treatment decisions and that there is a need for systemic chemotherapy, and surveillance monitoring. PMID- 21622215 TI - Glycosylated carriers for cell-selective and nuclear delivery of nucleic acids. AB - Targeted gene delivery via selective cellular receptors has been realized as a crucial strategy for successful gene therapy by maximizing therapeutic efficiency in target cells and minimizing systemic toxicity. The membrane carbohydrate binding proteins (membrane lectins) with different carbohydrate specificities are differentially expressed on the cellular and intracellular membranes of a number of cells. Their multiplicity, high affinity, and effective endocytosis after receptor binding as well as the biocompatibility of carbohydrate ligands endow them as potential ligands for glycosylated carriers in cell-selective delivery of nucleic acids. To achieve the in vivo application, glycosylated carriers/nucleic acid complexes have to fulfill certain conditions, including having a suitable size, minimal nonspecific interactions, low immunogenicity, and high uptake in target cells. Accordingly, the effective nuclear delivery of nucleic acids is the paramount important step for efficient gene transfer. This review summarizes the recent progress regarding application of glycosylated carriers for cell-selective and nuclear delivery of nucleic acids and their critical factors for efficient gene transfer. In addition, the development of new materials, such as carbon nanotubes, carbon nanospheres, and gold nanoparticles, as innovative carriers will be discussed with regards to glycosylation-mediated delivery of nucleic acids. PMID- 21622216 TI - Nuclear receptors CAR and PXR; therapeutic targets for cholestatic liver disease. AB - Cholestasis results in the intrahepatic retention of cytotoxic bile acids (BA) and it can thus lead to liver injury. Hydrophilic BA ursodeoxycholic acid (UDCA) is currently used to treat cholestasis but its efficacy is limited. Nuclear receptors are key regulators of various processes including metabolism of xeno- and endobiotics such as BA and drugs. Recent studies have made significant progress in elucidating the mechanisms which regulate the BA metabolism by nuclear receptors. The nuclear receptor FXR plays the role of master regulator of BA homeostasis and is a promising drug target for cholestatic liver disease. In addition to FXR, the nuclear receptors CAR and PXR function as sensors of toxic byproducts and regulator of BA homeostasis. Ligands for both receptors including phenobarbital have been used to treat cholestatic liver diseases before the mechanisms of these receptors were revealed. Novel compounds targeting CAR and PXR with specific effects and fewer side effects will therefore be useful for the treatment of cholestatic liver diseases. This article will review the current knowledge of the xenobiotic-sensing nuclear receptors CAR and PXR, while also discussing their potential role in the treatment of cholestatic liver diseases. PMID- 21622217 TI - Thymosin-alpha1 promotes the apoptosis of regulatory T cells and survival rate in septic mice. AB - Tregs are involved in immune disorder during sepsis; they can lead to a Th2 immune reaction. Their inhibitory effects can help alleviate inflammatory injury, but may also cause secondary immune inhibition. Thymosin-alpha1 is a polypeptide with powerful immunomodulatory activities. Current reports have shown that Thymosin-alpha1 conferred beneficial effects to septic patients. To explore the relationship between Thymosin-alpha1 and Tregs, in this study, we investigated the changing trend in CD4(+)CD25(+)Foxp3(+) T lymphocytes in a CLP septic mouse model. We also investigated the variation of apoptotic rate of CD4(+)CD25(+) T lymphocytes, cytokine variation, and change of model survival rate when Thymosin alpha1 intervening or not. We observed that the 72-h survival rate was improved, the percentage of CD4(+)CD25(+)Foxp3(+) T lymphocytes decreased and the apoptosis rate of CD4(+)CD25(+) T lymphocytes increased after intervention of Thymosin alpha1. At same time the expression of pro-inflammation cytokines IL-2, TNF-alpha and anti-inflammatory cytokines IL-10 and TGF-beta were regulated. In conclusion, Thymosin-alpha1 can effectively control the inflammatory response intensity and improve the 72-h survival rate of septic mice. Regulating Tregs may be another important role of Thymosin-alpha1 conditioning the immune reaction in sepsis. PMID- 21622218 TI - Sources of diversity in T cell epitope discovery. AB - CD8-positive T cells respond to small antigenic peptide fragments presented on class I major histocompatibility complexes (MHCs). Those specific T cell epitopes capable of precipitating a cellular immune response are either derived from (altered) self (i.e. they are autoimmune- or cancer-associated) or come from foreign sources (i.e. they are pathogen-associated). Identification of T cell epitopes provides elementary information that can be employed in technologies that monitor and predict the likely outcome of an immune response, as well as in therapeutic and vaccine development efforts. The coexistence between host and pathogen has largely driven the diversification of both their systems of immune surveillance and their antigenic determinants, respectively. In this review, we discuss the multitude of factors that introduce diversity to the T cell response from both sides of the host-pathogen interaction. Furthermore, we provide an overview of a variety of commonly employed methods and tools to characterize class I MHC restricted antigen presentation and recent endeavors towards the harmonization of reporting data concerning T cell responses. PMID- 21622219 TI - Effect of thymectomy on cellular immune function. AB - The present study was conducted to evaluate the effect of thymectomy during open heart surgery on immunological function of T lymphocytes in the treatment of children with congenital heart disease (CHD). No significant difference was found in the sjTREC level between pre-thymectomy and post- thymectomy in the non thymectomy group and the small partial resection group (P>0.05) However, the sjTREC level decreased from the pre-surgical level at 1 month (P<0.01) and 12 months (P<0.01) in the sub-total resection group. No differences were found in proportions of CD3, CD4 and CD8 lymphocytes, proliferative ability of lymphocytes and expression of IL-2, IL-4 and IFN-gamma after surgery between controls and three groups of patients (P>0.05). In the sub-total resection group, respiratory infection frequency (4.7+/-1.7 times) did not differ significantly from control group one year after surgery (P>0.05); however, mean days of anti-infection were significantly increased (P less than 0.01). In conclusion, sub-total thymectomy leads to a decrease in the sjTREC level in CHD children, whereas the function of peripheral mature T lymphocytes is normal. PMID- 21622220 TI - Serine phosphorylation of the Stat5a C-terminus is a driving force for transformation. AB - Persistent tyrosine phosphorylation of Stat3 and Stat5 is associated with oncogenic activity. Phosphorylation of the conserved tyrosine residue (pTyr) was long believed to be the only essential prerequisite to promote activation and nuclear translocation of Stat proteins. It has become evident, however, that post translational protein modifications like serine phosphorylation, acetylation, glycosylation as well as protein splicing and processing constitute further regulatory mechanisms to modulate Stat transcriptional activity and to provide an additional layer of specificity to Jak-Stat signal transduction. Significantly, most vertebrate Stat proteins contain one conserved serine phosphorylation site within their transactivation domains. This phosphorylation motif is located within a P(M)SP sequence. Stat transcription factor activity is negatively influenced by mutation of the serine to alanine. Moreover, it was shown for both Stat3 and Stat5 that their capacity to transform cells was diminished. This review addresses recent advances in understanding the regulation and the biochemical and biological consequences of Stat serine phosphorylation. In particular, we discuss their role in persistently activated Stat proteins for cancer research. PMID- 21622222 TI - Alternatively spliced tissue factor: discovery, insights, clinical implications. AB - Following the molecular cloning of human Tissue Factor (TF) in mid-1980's, great strides have been made in the understanding of TF biology, TF's crucial roles in the initiation of blood coagulation and embryonic development, and TF's contribution to the pathobiology of various disease states. The 21st century brought about a rather unexpected turn in the "TF journey"--a few years back it was reported that the TF gene produces not one, but two proteins with distinct structural and functional characteristics. The so-called "full-length TF" (flTF) a much-studied integral membrane glycoprotein long presumed to be, and experimentally handled as "the TF" in hundreds of laboratories around the world - is now known to be one of the two TF forms naturally occurring in humans as well as mice. The other, recently discovered form is termed alternatively spliced TF (asTF) which, unlike flTF, lacks a transmembrane domain and can thus be secreted. In this review, we summarize the literature on asTF by discussing asTF's biologic roles as they are currently understood, tackling a number of questions pertaining to asTF's evident and proposed biologic properties, and briefly covering the emerging field of regulated TF pre-mRNA processing. PMID- 21622221 TI - G protein-coupled receptor kinases in normal and failing myocardium. AB - Heart failure (HF) is the end stage of many underlying cardiovascular diseases and is among the leading causes of morbidity and mortality in industrialized countries. One of the striking characteristics of HF is the desensitization of G protein-coupled receptor (GPCR) signaling, particularly the beta-adrenergic receptor (betaAR) system. GPCR desensitization is initiated by phosphorylation by GPCR kinases (GRKs), followed by downregulation and functional uncoupling from their G proteins. In the heart, the major GRK isoforms, GRK2 and GRK5, undergo upregulation due to the heightened sympathetic nervous system activity that is characteristic of HF as catecholamine levels increase in an effort to drive the failing pump. This desensitization leads to the distinctive loss of inotropic reserve and functional capacity of the failing heart. Moreover, GRK2 and GRK5 have an increasing non-GPCR interactome, which may play critical roles in cardiac physiology. In the current review, the canonical GPCR kinase function of GRKs and the novel non-GPCR kinase activity of GRKs, their contribution to the pathogenesis of cardiac hypertrophy and HF, and the possibility of GRKs serving as future drug targets will be discussed. PMID- 21622223 TI - Light-induced water oxidation in photosystem II. AB - The photosystem II core complex (PSIIcc) is the key enzyme of oxygenic photosynthesis, as it catalyzes the light-induced oxidation of water to form dioxgyen and protons. It is located in the thylakoid membrane of cyanobacteria, algae, and plants and consists of 20 protein subunits binding about 100 cofactors. In this review, we discuss what is presently known about the "donor side" of PSIIcc, covering the photosynthetic reaction center and the water oxidase part. The focus is on the catalytic Mn4Ca cluster and its protein environment. An attempt is made to connect recent crystallographic data (up to 2.9 A resolution) with the wealth of information about Nature's water oxidation device from spectroscopic, biochemical and theoretical work. PMID- 21622224 TI - Structural evidence of anti-atherogenic microRNAs. AB - Our research attempted to address two important questions--how microRNAs modulate atherogenic inflammatory genes from a panoramic viewpoint and whether their augmented expression results from reduced microRNAs suppression. To resolve these knowledge gaps, we employed a novel database mining technique in conjunction with statistical analysis criteria established from experimentally verified microRNAs. We found that the expression of 33 inflammatory genes up-regulated in atherosclerotic lesions contain structural features in the 3'UTR of their mRNAs for potential microRNAs regulation. Additionally, the binding features governing the interactions between the microRNAs and the inflammatory gene mRNA were statistically identical to the features of experimentally verified microRNAs. Furthermore, 21 of the 33 inflammatory genes (64%) were targeted by highly expressed microRNAs and 10 of these (48%) were targeted by a single microRNA, suggesting microRNA regulation specificity. Supplementing our findings, 7 out of the 20 unique microRNAs (35%) were previously confirmed to be down-regulated when treated with pro-atherogenic factors. These results indicate a critical role of anti-inflammatory microRNAs in suppressing pro-atherogenic inflammatory gene expression. PMID- 21622225 TI - Structure and regulation of the c-Fes protein-tyrosine kinase. AB - The c-Fes protein-tyrosine kinase is the normal cellular ortholog of several avian and feline retroviral oncoproteins. Unlike its transforming viral counterparts, c-Fes tyrosine kinase activity is tightly regulated in vivo through a mechanism involving coiled-coil oligomerization domains and other unique structural features found in its long N-terminal region. This review is focused on the regulatory features and structural biology of c-Fes, which has been implicated in normal cellular growth regulation, the innate immune response, and tumorigenesis. PMID- 21622226 TI - Allosteric activation of coagulation factor VIIa. AB - Coagulation factor VIIa (FVIIa) is present at subnanomolar concentration and represents a small percentage of the total amount of FVII in the circulation. FVIIa is poised to initiate blood clotting when it encounters its pivotal cofactor tissue factor (TF) which becomes exposed to blood upon vascular rupture. The requirement for complex formation with TF in order for FVIIa to express procoagulant activity ensures thrombin and fibrin generation at the right time and place. Thus TF acts as a guardian of safety of paramount importance to blood coagulation by providing localization to the site of injury and at the same time inducing maturation of zymogen-like free FVIIa to the active cofactor-bound enzyme. This review gives an account of the accumulated knowledge about the structure, function and TF dependence of FVIIa to arrive at a plausible allosteric mechanism by which TF induces maturation of the active conformation of FVIIa. PMID- 21622227 TI - Escherichia coli Y family DNA polymerases. AB - DNA damage is ubiquitous, arising from both environmental and endogenous sources. All organisms have evolved multiple pathways to respond to DNA damage and maintain genomic integrity. Escherichia coli possesses two DNA polymerases, pol IV and pol V, that are members of the Y family. These polymerases are characterized by their specialized ability to copy damaged DNA as well as their relatively low fidelity on undamaged DNA. Pol IV and pol V are regulated by the SOS response to DNA damage and by their multiple interactions with other proteins. These two Y family DNA polymerases copy DNA damaged by distinct agents. Pol IV is capable of replicating DNA containing N(2)-dG adducts, while pol V bypasses abasic sites and thymine-thymine dimers, which result from exposure to UV radiation. In addition to their roles in copying damaged DNA, the two Y family DNA polymerases in E. coli act in regulation of DNA replication and contribute to bacterial mutagenesis in response to cellular stress. PMID- 21622228 TI - Tocotrienol combination therapy results in synergistic anticancer response. AB - Vitamin E represents a family of compounds that is divided into two subgroups called tocopherols and tocotrienols, which act as important antioxidants that regulate peroxidation reactions and control free-radical production within the body. However, many of the biological effects of vitamin E are mediated independently of its antioxidant activity. Although tocopherols and tocotrienols have the same basic chemical structure characterized by a long phytyl chain attached to a chromane ring, only tocotrienols display potent anticancer activity, by modulating multiple intracellular signaling pathways associated with tumor cell proliferation and survival, and combination therapy with other chemotherapeutic agents result in a synergistic anticancer response. Combination therapy is most effective when tocotrienols are combined with agents that have complementary anticancer mechanisms of action. These findings strongly suggest that the synergistic antiproliferative and apoptotic effects demonstrated by combined low dose treatment of gamma-tocotrienol with other chemotherapeutic agents may provide significant health benefits in the prevention and/or treatment of breast cancer in women, while at the same time avoiding tumor resistance and toxic side effects associated with high dose monotherapy. PMID- 21622229 TI - Tissue factor and factor VIIa cross-species compatibility. AB - Knowledge about species compatibility is crucial for proper interpretation of data from in vivo experiments with human proteins in pharmacological models and of data from cross-species in vitro experiments. Information about the cross species compatibility of tissue factor (TF) and coagulation factor (F) VII (FVII) has accumulated since the early history of coagulation research. Many observations were connected to the introduction and development of the prothrombin time (PT) assay where fibrin clot formation was observed when tissue extracts of different origins were added to recalcified human or non-human plasmas. Studies on cross-species TF-FVIIa compatibility entered into a new area with the cloning and recombinant expression of TF and FVII from a number of species as well as with the possibility of specific amino acid substitution. TF and/or FVIIa from cattle, dog, rabbit, mouse, rat and zebrafish have been purified and characterized in varying detail. In addition to adding knowledge about the species-specific TF-FVIIa interactions, cross-species studies often reveal information which adds to the general view of the structural and functional properties of the human TF-FVIIa complex. This review briefly outlines the features of human TF and FVIIa, their intermolecular interactions, and the biological effects of TF-FVIIa complex formation and compares this information to findings obtained in studies addressing TF or FVIIa of non-human origin. By examples we point to difficulties which may arise from the transcendence across species borders and how some cross-species data have advanced our understanding of the structure and function of the human TF-FVIIa complex. PMID- 21622230 TI - Functional characterization of (pro)renin receptor in association with V-ATPase. AB - The (pro)renin receptor ((P)RR) is a unique molecule that binds prorenin and renin in tissues, not only leading to their activation, but also inducing intracellular signaling. As a key player in the local renin-angiotensin system, (P)RR activation plays an important role in the development of cardiac fibrosis and proteinuria in hypertension and diabetes. Intriguingly, the fragment (P)RR is also called ATP6AP2 because it has been shown to be associated with vacuolar-type H(+)-ATPase (V-ATPase). The V-ATPase is a multi-subunit proton pump involved in diverse and fundamental cellular processes, including receptor-mediated endocytosis, processing of proteins and signaling molecules, membrane sorting and trafficking, and activation of lysosomal enzymes. The role of (P)RR in the function of the V-ATPase is implicated in the previous findings and vigorously investigated in the recent studies. Furthermore, the novel function of the (P)RR as an adaptor protein between the Wnt receptor complex and the V-ATPase was discovered. Thus, the (P)RR is a multi-functional molecule that shows the complex structure and behaviour. This review highlights the current insights and the future perspectives in research regarding the (P)RR and mammalian V-ATPase. PMID- 21622231 TI - Tyrosine kinase 2 (TYK2) in cytokine signalling and host immunity. AB - The Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signal transduction pathway is essential to transmit signals from transmembrane receptors to the nucleus in order to alter gene expression programs and to respond to extracellular cues. Tyrosine kinase 2 (TYK2) was the first member of the JAK family that was identified within a screen for molecules complementing human cell lines mutant for interferon (IFN) responses. During the last decades biochemical studies and gene-targeted mice uncovered the crucial role of TYK2 in immunity. Tyk2-deficient mice are viable and fertile but display multiple immunological defects, most prominently high sensitivity to infections and defective tumour surveillance. In contrast, absence of TYK2 results in increased resistance against allergic, autoimmune and inflammatory diseases. In support of these data, the only patient with TYK2 deficiency described so far displays high serum immunoglobulin E (IgE) levels and increased sensitivity to infectious diseases. Furthermore, numerous genome-wide association studies in humans propose a link between TYK2 genetic variants and several autoimmune diseases, inflammatory diseases and tumours. Thus, TYK2 appears as an attractive target for therapeutic intervention. Future work will be required to further delineate structure-function relationships and to fully understand the involvement of TYK2 in immune regulatory networks. PMID- 21622232 TI - Interactions between endothelial selectins and cancer cells regulate metastasis. AB - The selectins: E-selectin, P-selectin, and L-selectin are adhesion molecules that are crucial for binding of circulating leukocytes to vascular endothelium during the inflammatory response to injury or infection. Accumulated evidence indicates that selectins regulate adhesion of circulating cancer cells to the walls of blood vessels. Selectin ligands are transmembrane glycoproteins expressed on leukocytes and cancer cells that promote bond formations with selectins to mediate inflammatory processes. Selectins and selectin ligands also participate in signal transduction to regulate diverse cellular functions. Sialyl Lewis X (sLe(x)) and sialyl Lewis A (sLe(a)) tetrasaccharides are carbohydrate motifs displayed on protein or lipid scaffolds that are critical components of functional selectin ligands. Selectin binding to sLe(x) and sLe(a) present on colon, gastric, bladder, pancreatic, breast, and prostate carcinomas enhances distant organ metastasis. High expression of sialyl Lewis ligands on these cancers is significantly correlated with a poor post-operative prognosis. This review will focus on the roles of E-selectin and P-selectin in cancer progression. Understanding the role of selectins in cancer supports the development of novel selectin-based therapies to control metastasis. PMID- 21622233 TI - The DFNB1 subtype of autosomal recessive non-syndromic hearing impairment. AB - Inherited hearing impairment is a frequent and highly heterogeneous condition. Among the different subtypes of autosomal recessive non-syndromic hearing impairment, DFNB1 is remarkable for its high frequency in most populations. It is caused by mutations in the coding region or splice-sites of the GJB2 gene, or by mutations affecting regulatory sequences that are essential for the expression of this gene. GJB2 encodes connexin-26, a protein component of intercellular gap junctions, which play crucial physiological roles in the cochlea. Because of its high frequency, DFNB1 hearing impairment has received continued attention from researchers along the years, resulting in a wealth of data that is unparalleled among these disorders. Here we review our current knowledge on the genetic, molecular, and phenotypic aspects of this subtype of hearing impairment. PMID- 21622234 TI - Current and nano-diagnostic tools for dengue infection. AB - Dengue is one of the infectious diseases that is widespread over global regions with yearly occurrence of epidemics and could be deadly in some cases. Thus the developments of rapid and specific diagnostic tools which can achieve early detection of dengue infection for disease control during epidemic situations and before complications occur are deemed highly desirable. This paper describes the current and advanced methods for diagnosis of dengue infection and discusses the advantages and disadvantages of these methods in terms of their analytical performances and clinical applicabilities. The current methods discussed herein include enzyme-linked immunosorbent assay and reverse transcriptase polymerase chain reaction. In addition, recent instrumental methods such as quartz crystal microbalance, surface plasmon resonance, photonic crystal and electrochemical impedance spectroscopy have shown promising results. Interesting developments in detection of dengue infection using nanosized materials including liposomes, nanowires and nanopores, coupled to conventional fluorescence, potentiometry and voltammetry methods are also described and could possibly point the way forward for the development of inexpensive diagnostic tools for use at point-of-care and in events of epidemic scale. PMID- 21622236 TI - The good, the bad and the ugly. Macrophages/microglia with a focus on myelin repair. AB - A feature of most neurological disorders is demyelination, whereby myelin is lost from axons partly through stripping by macrophages/microglia. Spontaneous remyelination by oligodendrocytes that mature from oligodendrocyte precursor cells occurs following demyelination, even in the chronic inflammatory disorder of the central nervous system, multiple sclerosis. If remyelination does not occur or is prevented, then one consequence besides the loss of saltatory nerve conduction is the degeneration of axons. Thus, promoting remyelination is a desired result. In this article, we review the data that despite a reputation as "bad" factors for CNS wellbeing, including the promotion of neuroinflammation and demyelination, some aspects of macrophages/microglia activity are indeed "good", and can engender repair from the "ugly" phenomenon of demyelination. We discuss factors that help promote the benefits of macrophages/microglia activity for remyelination. PMID- 21622237 TI - Mechanisms and consequences of microglial responses to peripheral axotomy. AB - Microglia respond rapidly to injury of peripheral nerve axons (axotomy). This response is integrated into the responses of the injured neurons, i.e. processes for neuron survival, axon regeneration and restoration of target contact. The microglial response is also integrated in changes in presynaptic terminals on axotomized motor or autonomic neurons and in changes in the central terminals of peripherally axotomized sensory neurons. Microglia also has an established role in interacting with astrocytes to shape their response to peripheral axotomy. Axotomy models in mice have demonstrated a role for microglia in regulating the entry of lymphocytes into motor nuclei or sensory areas following peripheral axotomy. Whether this is a universal component of peripheral nerve injury remains to be determined. Under certain circumstances, microglia activated by axotomy are major contributors to CNS pathology, e.g. in models of neuropathic pain. However, the general roles played by microglia after peripheral nerve injury are still incompletely understood. Early proposals that the microglial reaction to peripheral nerve injury is preparatory for the eventuality of neuron degeneration may still have relevance. PMID- 21622235 TI - Effects of opioids, cannabinoids, and vanilloids on body temperature. AB - Cannabinoid and opioid drugs produce marked changes in body temperature. Recent findings have extended our knowledge about the thermoregulatory effects of cannabinoids and opioids, particularly as related to delta opioid receptors, endogenous systems, and transient receptor potential (TRP) channels. Although delta opioid receptors were originally thought to play only a minor role in thermoregulation compared to mu and kappa opioid receptors, their activation has been shown to produce hypothermia in multiple species. Endogenous opioids and cannabinoids also regulate body temperature. Mu and kappa opioid receptors are thought to be in tonic balance, with mu and kappa receptor activation producing hyperthermia and hypothermia, respectively. A particularly intense research focus is TRP channels, where TRPV1 channel activation produces hypothermia whereas TRPA1 and TRPM8 channel activation causes hyperthermia. The marked hyperthermia produced by TRPV1 channel antagonists suggests these warm channels tonically control body temperature. A better understanding of the roles of cannabinoid, opioid, and TRP systems in thermoregulation may have broad clinical implications and provide insights into interactions among neurotransmitter systems involved in thermoregulation. PMID- 21622238 TI - Microglia--friend or foe. AB - Microglia, as the immune effectors in the central nervous system, respond to pathological conditions and participate in the initiation and progression of neurological disorders such as inflammation and brain tumor by releasing potential neurotrophic or cytotoxic molecules, presenting the antigen to T cell and interacting with brain tumor. Evidences also suggest that microglia are capable of promoting or inhibiting the proliferation and differentiation of neural stem cells by secreting series of biologically active molecules. In this review, we focus on three aspects-inflammation, neurogensis and brain tumor to illustrate the multi-faceted activities of microglia in the normal and pathologic brain. PMID- 21622239 TI - Role of microglia in the process of inflammation in the hypoxic developing brain. AB - The developing brain is susceptible to hypoxic damage because of its high oxygen and energy requirements. Hypoxia-induced inflammatory response has been recognized as one of the main culprits in the development of hypoxic brain injury. In this regard, a hallmark feature is microglial activation which results in overproduction of inflammatory cytokines, free radicals and nitric oxide. Concomitantly, activated microglia exhibit enhanced expression of ion channels such as Kv1.2, Kv1.1 and Nav which further promote the release of inflammatory cytokines, chemokines and reactive oxygen species. Through the above-mentioned inflammatory mediators, activated microglia induce neuronal loss, axonal damage and oligodendroglial death along with myelination disturbances. Our recent studies have extended that tumor necrosis factor-alpha, interleukin-1beta, monocyte chemoattractant protein-1 and macrophage colony stimulating factor produced by activated microglia are linked to the pathogenesis of periventricular white matter damage in the hypoxic brain. It is envisaged that a better understanding of the interactions between microglia and neurons, axons and oligodendrocytes is key to the development of effective preventive and therapeutic strategies for mitigation of hypoxic brain injury. PMID- 21622240 TI - Stem cells cardiac differentiation in 3D systems. AB - Cardiac regeneration requires a complex cascade of events. Stem cell therapy and tissue engineering are newly emerging tools with promising potential for recover or replace of damaged cardiac tissue. There are many factors, most of them still no clarified, that limit the effectiveness of these treatments and their translation to the clinic. Cells should graft, survive and functionally integrate to the target organ in order to have a chance to restore its function. As in original tissues, a complex and well defined set of signals, many of them coming from the extracellular matrix, is required for normal cell physiology. Biomaterials science gives us important tools to build this extracellular matrix. Functionalized 3D systems can provide the correct environment and act as a delivery system for genes or gene products, guiding the therapeutic cells to the functional phenotype. PMID- 21622241 TI - Histopathology and outcome of acute humoral rejection in renal allografts. AB - Our purpose was to see if histopathologic features of acute antibody-mediated rejection (AMR) in renal allografts have prognostic value; and to compare two year graft survival with and without additional therapy with plasmapheresis and intravenous immunoglobulin (IVIG). We reviewed renal allograft biopsies taken within the first 6 months after transplant from patients with C4d positive AMR, performed between January 2000 to December 2005 (n=57). We formed two groups: Group 1: biopsied between 2003 and 2005 (n=26), when C4d staining was routinely performed and option for plasmapheresis and IVIG was available; Group 2: biopsied between 2000 and 2002 (n=31), retrospectively found to be C4d positive. Patients whose biopsies showed cortical necrosis or arterial fibrinoid necrosis had early graft loss. Other histopathologic features did not statistically correlate with graft loss. Overall, additional plasmapheresis/IVIG treatment did not show convincing improvement in graft survival or function at 2 years post-transplant, but all six patients with thrombotic microangiopathy (TMA) who received plasmapheresis/IVIG had functioning grafts at two-year follow-up. PMID- 21622242 TI - Desensitization across the HLA barrier in kidney transplantaion. AB - The number of highly sensitized patients on the transplant waiting list continues to rise. HLA allosensitization has profound effects on the wait time to transplant, rejection rates, and long-term outcomes. While technological advances with high sensitivity and specificity have facilitated the detection of donor specific antibodies, there is no consensus on diagnostic and prognostic values of these tests. Recently, multiple desensitization protocols have been developed that comprise high dose IVIG, low dose IVIG with plasmapheresis, rituximab and more novel agents including Bortezomib. Although these preconditioning protocols have led to successful transplantation of sensitized patients, long-term outcomes are limited and suboptimal. Randomized clinical trials are needed to determine optimal treatment and monitoring strategies in patients that are highly sensitized across the HLA barrier. PMID- 21622243 TI - Finding the root of the problem: the quest to identify melanoma stem cells. AB - Melanoma is an exceptionally aggressive cancer with limited treatment options. As such, the idea that a minority of tumor cells, termed melanoma stem cells, are actually responsible for the progression of the disease offers up new possibilities for targeted therapies. However, reliable identification of these melanoma stem cells is complicated by the lack of clearly defined markers to distinguish them from the general tumor cell population. Additionally, there is evidence that under permissive conditions, a high proportion of melanoma cells are capable of forming tumors in mice. This review summarizes a number of the possible markers being considered for identifying melanoma stem cells, the potential role of transcription factors that regulate pluripotency and stem cell maintenance in melanoma, and evidence that may undermine the applicability of the cancer stem cell hypothesis to melanoma. PMID- 21622244 TI - Endothelial dysfunction in hypertension: the role of arginase. AB - Essential hypertension is the leading risk factor for mortality worldwide, accountable for 13% of deaths globally. Despite numerous therapies available uncontrolled hypertension is still very prevalent today and a large subset are shown to have treatment resistant hypertension. Several cardiovascular diseases including hypertension result in endothelial dysfunction and inflammation. Once thought of as a passive barrier between blood flow and tissue the endothelium is now considered a main hub for maintaining vascular tone, structure and haemostasis. Several pathways occur in the endothelium that can result in dysfunction and altered vascular stasis. Such pathways include the impairment of the vasodilator nitric oxide (NO), increases in pro-inflammatory pathways such as ROS (reactive oxygen species) production and also recent reports suggest that the enzyme arginase, associated with the L-arginine-urea cycle, may be an important factor that is increased in hypertension. These pathways may offer alternative mechanisms to treat the complications associated with hypertension rather than the conventional therapies that aim to lower blood pressure. PMID- 21622245 TI - Thymus-bound: the many features of T cell progenitors. AB - T cells are unique in that they begin their development as a progenitor within the bone marrow but complete their differentiation within the thymus. Furthermore, long-term T-lymphopoiesis requires a continuous supply of thymus bound progenitors derived from the bone marrow. The critical role for T cells is clearly observed in individuals with genetic or acquired immunodeficiencies or those having undergone hematopoietic stem cell transplantation. Here, we review the work done by several groups aimed at characterizing the earliest T-lineage progenitors (ETPs), in mouse and human, found within the thymus, in addition to the long-sought after thymus-colonizing progenitor, which makes its journey from the bone marrow via the bloodstream into thymus. The characterization of these progenitors may herald therapeutic insight into the restoration of T cells in immunodeficient individuals. PMID- 21622246 TI - Role of miRNA in distinguishing primary brain tumors from secondary tumors metastatic to the brain. AB - Cancer is the result of complex processes that involve multiple molecular alterations. The understanding of such complexity has been improved by the advent of a new class of small, noncoding RNA gene products known as microRNAs (or miRNAs). miRNAs play an essential role in cancer development and progression by modulating gene expression binding to target mRNA, causing either mRNA degradation or translation inhibition. Several studies have shown that miRNAs can act either as tumor suppressors or as oncogenes, and that measurement of miRNA expression in malignancies may have diagnostic and prognostic implications. Beyond these valuable features, miRNAs could be potentially used in the future as innovative and targeted therapeutics. Recent in vitro and expression profiling studies have identified that specific miRNAs are directly involved in brain carcinogenesis and in the metastatic process. This review focuses on metastasis related miRNAs and on the role of miRNAs in distinguishing between primary and metastatic brain tumors. In clinical practice, miRNAs could represent a promising new class of cancer biomarkers in the diagnosis and management of brain neoplastic lesions. PMID- 21622247 TI - Origins of the concept of store-operated calcium entry. AB - The concept of capacitative or store-operated calcium entry, a process by which the release of stored calcium signals the opening of plasma membrane calcium channels, has its roots in the late 1970's, and was formalized in 1986. This short introduction to the current volume of Frontiers in Bioscience briefly summarizes the early experimental work that led to the idea of store-operated calcium entry, and provided the initial proofs for it. PMID- 21622248 TI - Pathogenetic and therapeutic implications of the histamine H4 receptor in inflammatory skin diseases and pruritus. AB - Chronic inflammatory skin diseases such as atopic dermatitis (AD) are clinically characterized by erythematous and pruritic skin lesions, immunologically mediated by an inflammatory infiltrate consisting of T-cells, antigen presenting cells (APC) and eosinophilic granulocytes. Histamine levels are increased in lesions of inflammatory skin diseases. It is likely that histamine also plays a pathogenetic role since various relevant cell types such as T-cells and APC express functional histamine receptors. However, therapeutic blockade of the histamine H1 and H2 receptor is inefficient at least in the treatment of atopic dermatitis. We summarize here current data on the role of the recently described histamine H4 receptor (H4R) in chronic inflammatory skin diseases. The H4R is functionally expressed on relevant cell types such as T-cells, APC and keratinocytes. In murine models of contact hypersensitivity and pruritus, H4R blockade had significant in vivo effects. Taken together, several lines of evidence suggest a role of the H4R in chronic inflammatory skin disease and the H4R might be a therapeutic target for diseases such as AD. PMID- 21622249 TI - Podocyte dysfunction in aging--related glomerulosclerosis. AB - We review podocyte molecular structure and function, consider the underlying mechanisms related to podocyte dysfunction and propose that podocyte dysfunction be considered in the evaluation and management of age-associated glomerulosclerosis. With aging, progressive sympathetic activation, increased intrarenal renin-angiotensin system (RAS) activity, endothelin system and oxidative stress and reduced nitric oxide (NO)-availability can damage podocytes. Apoptosis and proliferation are the principal podocyte changes following injury with the latter leading to sclerosis and loss of nephrons. Podocyte loss can be evaluated by either determining their average number in biopsed glomeruli or by estimating podocyte number or their associated molecules in urine sediment. Podocyturia may be considered a marker of active glomerular disease. Preliminary data suggest that antiadrenergic drugs, angiotensin converting enzyme (ACE) inhibitors, RAS blocking drugs, endothelin system inhibitors and reduced oxidative stress can protect podocytes. Thus podocytes appear to play an important role in the pathogenesis, evaluation and therapy of age related glomerulosclerosis. PMID- 21622250 TI - New role of glutamate as an immunoregulator via glutamate receptors and transporters. AB - Accumulating evidence suggests that the amino acid glutamate (Glu) may play a role in mediating immune function. The demonstration of Glu receptors (GluR) and Glu transporters (GluT) on a variety of immune cells suggests that Glu has a functional role in immunoregulation well beyond its role as a neurotransmitter. The extracellular Glu concentration plays a key role in the regulation of GSH synthesis in immune cells via 2 key GluTs (i.e., Xc- and X-AG systems). Emerging evidence also suggests a role of Glu as signaling molecule in immune cells via ionotropic GluRs (iGluRs) and metabotropic GluRs (mGluRs). In vitro, extracellular Glu concentration has been shown to exert a dose-dependent regulation on lymphocyte activation/proliferation. Specifically, Given the exceedingly high intestinal Glu concentration, these finding are suggestive of a potential role for Glu in modulating immune function and promoting tolerance in the gut associated lymphoid tissues. PMID- 21622251 TI - Neural regulation of bone, marrow, and the microenvironment. AB - Bone marrow niches are specialized microenvironments comprising a heterogeneous population of cells that support and regulate hematopoietic stem and progenitor cells. Considerable advances made in the field of hematopoiesis reveal a cross talk between cells, cytokines and neurotransmitters of the hematopoietic, immune and skeletal systems. Dynamic modulation and regulation of stem cells and their niches in response to internal and external stimuli are essential for homeostasis, host defense and repair. This review presents evidence to substantiate stem cell regulation via the "brain-bone-blood triad" beginning at the embryonic stage and continuing to adulthood. PMID- 21622252 TI - MicroRNA-122: a therapeutic target for hepatitis C virus (HCV) infection. AB - Hepatitis C virus infection is the main cause of liver disease worldwide, often leading to chronic hepatitis. Recent studies have demonstrated that miRNA-122, a liver-specific miRNA, is required for HCV replication in hepatocytes by its binding to the 5' UTR of HCV. Down-regulation of miRNA-122 in vitro and in vivo has led to significant inhibition of viral replication. In the present article, we report the major recent findings on the potential therapeutic role of anti miRNA-122 molecules. PMID- 21622253 TI - Bone phenotypes of P2 receptor knockout mice. AB - The action of extracellular nucleotides is mediated by ionotropic P2X receptors and G-protein coupled P2Y receptors. The human genome contains 7 P2X and 8 P2Y receptor genes. Knockout mice strains are available for most of them. As their phenotypic analysis is progressing, bone abnormalities have been observed in an impressive number of these mice: distinct abnormalities in P2X7-/- mice, depending on the gene targeting construct and the genetic background, decreased bone mass in P2Y1-/- mice, increased bone mass in P2Y2-/- mice, decreased bone resorption in P2Y6-/- mice, decreased bone formation and bone resorption in P2Y13 /- mice. These findings demonstrate the unexpected importance of extracellular nucleotide signalling in the regulation of bone metabolism via multiple P2 receptors and distinct mechanisms involving both osteoblasts and osteoclasts. PMID- 21622254 TI - In vitro studies of early cardiac remodeling: impact on contraction and calcium handling. AB - Cardiac remodeling, hypertrophy, and alterations in calcium signaling are changes of the heart that often lead to failure. After a hypertrophic stimulus, the heart progresses through a state of compensated hypertrophy which over time leads to decompensated hypertrophy or failure. It is at this point that a cardiac transplant is required for survival making early detection imperative. Current experimental systems used to study the remodeling of the heart include in vivo systems (the whole body), isolated organ and sub-organ tissue, and the individual cardiac muscle cells and organelles.. During pathological remodeling there is a derangement in the intracellular calcium handling processes. These derangements are thought to lead to a dysregulation of contractile output. Hence, understanding the mechanism between remodeling and dysregulation is of great interest in the cardiac field and will ultimately help in the development of future treatment and early detection. This review will center on changes in contraction and calcium handling in early cardiac remodeling, with a specific focus on findings in two different in vitro model systems: multicellular and individual cell preparations. PMID- 21622255 TI - The function of histamine receptor H4R in the brain revealed by interaction partners. AB - The histamine H4 receptor is mainly expressed in haematopoietic cells, hence is linked to inflammatory and immune system conditions. It has been recently discovered that the receptor is expressed also in the mammalian central nervous system (CNS), but its role in the brain remains unclear. We address the potential functions of the histamine H4 receptor in the human brain using a 'guilty by association' logic, by close examination of protein-protein functional associations networks in the human proteome. PMID- 21622256 TI - Protein palmitoylation in protozoan parasites. AB - Palmitoylation plays an important role in the regulation of the localization and function of the modified protein. Although many aspects of protein palmitoylation have been identified in mammalian and yeast cells, little information is available of this modification in protozoan parasites. Protein palmitoylation has been described for a few set of proteins in E.tenella, P. falciparum, T. gondii, G. lamblia and T. cruzi. Interestingly, in all these parasites palmitoylated proteins appears to be involved in vital processes such as invasion and motility. In addition, most of these parasites contain in their genomes genes that encode for putative palmitoyl-acyl transferases, the enzymes catalyzing the palmitoylation reaction. Although protein palmitoylation could be playing key roles in invasion and motility in a variety of parasites, little is known about this important reversible modification of proteins that typically plays a role in membrane tethering. As such, this review will focus on the main features of protein palmitoylation as well as provide an overview of the state of knowledge of this modification in protozoan parasites. PMID- 21622257 TI - Lactoferrin and cancer in different cancer models. AB - Lactoferrin (Lf) is a multifunctional protein and an essential element of innate immunity. Cancer is a major killer in today's world accounting for around 13% of all deaths according to the World Health Organisation (W.H.O.). The five most common forms of cancer include lung, colorectal, stomach, liver and breast cancer. Lactoferrin is a natural forming iron-binding glycoprotein with antibacterial, antioxidant and anti-carcinogenic effects. It is produced in exocrine glands and is secreted in many external fluids as a first line of defence. Lactoferrin also has the capacity to induce apoptosis and inhibit proliferation in cancer cells as well as restore white and red blood cell levels after chemotherapy. This review focuses on the therapeutic effect bovine sourced lactoferrin has on various forms of cancer in various models. It also focuses on the benefits of 3D in vitro cell culture. 3D cell culture has vast advantages over 2D models including demonstration of realistic therapeutic results and heightened resistance that 2D models fail to display. PMID- 21622258 TI - Fire within the vessels: interactions between blood cells and inflammatory vascular injury. AB - Inflammation involves multiple molecular, humoral and cellular mechanisms that aim to protect the body from injury or infection by the tight orchestration of immune cell trafficking, interactions and activation. Platelets and neutrophils are major players in the initial thromboinflammatory response by rapidly responding to a variety of danger signals. When the intensity, timing or locations of these responses are uncontrolled, however, they can trigger localized or systemic injury. Studies over the past decades have revealed that during the normal inflammatory response, blood elements frequently interact with each other to form heterotypic aggregates. The formation of these aggregates within blood vessels, in turn, underlies injury in several models of acute inflammation and can in some instances lead to death. These phenomena are likely to have a bigger contribution to the outcome of inflammatory processes than previously expected, not only in acute scenarios but also in those that involve chronic vascular damage, such as atherosclerosis. We will review here the molecular mediators of these interactions and their consequences in the context of cardiovascular injury. PMID- 21622259 TI - P2 receptor expression, signaling and function in osteoclasts. AB - Skeletal development and bone remodeling depend on the coordinated activity of osteoblasts and osteoclasts, which are responsible for bone formation and resorption, respectively. Mature osteoclasts result from the fusion of precursor cells, and they are large, multinucleated, highly specialized cells. Cellular release of ATP and UTP occurs in response to a variety of stimuli including mechanical stimulation, which occurs in the bone environment. ATP and UTP or their metabolites can then act on P2 receptors in the plasma membrane to induce various responses in bone cells. The influence of these receptors on osteoclast physiology and bone physiology in general is beginning to be understood, but much work is still required. This review focuses on P2 receptors in osteoclasts, their expression, signaling and function in the regulation of osteoclast formation, resorptive activity and survival. PMID- 21622260 TI - RAGE during infectious diseases. AB - The receptor for advanced glycation end products (RAGE) is a multiligand receptor that is expressed at high levels in the lungs. The emerging concept of pattern recognition involves RAGE and Toll-like receptors (TLRs) in sensing not only "pathogen-associated molecular patterns" (PAMPs) but also (endogenous) damage associated molecular patterns (DAMPs). Infection is associated with the release of these endogenous proteins, such as high-mobility group box-1 (HMGB1) and S100A12. Engagement of RAGE by its diverse ligands results in receptor-dependent signaling and activation of NF-kappaB. Furthermore, RAGE acts as an endothelial adhesion receptor for leukocyte integrins and promotes leukocyte recruitment. Inhibition of RAGE signaling reduces inflammatory responses in several (non infectious) models as well as in infectious models of cecal ligation and puncture and S. pneumoniae pneumonia. Importantly, RAGE signaling inhibition increased bacterial outgrowth and dissemination in an E. coli abdominal sepsis model. This review describes experimental studies that provide further insight into the role of RAGE and its ligands in host defense during clinically important infections, which eventually may contribute to better therapies against specific pathogens. PMID- 21622261 TI - A-type lamins and Hutchinson-Gilford progeria syndrome: pathogenesis and therapy. AB - Lamin A and lamin C (A-type lamins, both encoded by the LMNA gene) are major components of the mammalian nuclear lamina, a complex proteinaceous structure that acts as a scaffold for protein complexes that regulate nuclear structure and function. Abnormal accumulation of farnesylated-progerin, a mutant form of prelamin A, plays a key role in the pathogenesis of the Hutchinson-Gilford progeria syndrome (HGPS), a devastating disorder that causes the death of affected children at an average age of 13.5 years, predominantly from premature atherosclerosis and myocardial infarction or stroke. Remarkably, progerin is also present in normal cells and appears to progressively accumulate during aging of non-HGPS cells. Therefore, understanding how this mutant form of lamin A provokes HGPS may shed significant insight into physiological aging. In this review, we discuss recent advances into the pathogenic mechanisms underlying HGPS, the main murine models of the disease, and the therapeutic strategies developed in cellular and animal models with the aim of reducing the accumulation of farnesylated-progerin, as well as their use in clinical trials of HGPS. PMID- 21622262 TI - C-type lectins in immunity to Mycobacterium tuberculosis. AB - Tuberculosis (TB) remains one of the leading causes of death due to a single infectious agent, Mycobacterium tuberculosis, with nearly 2 million deaths per year (1). Most individuals exposed to the bacillus develop a nonpathological form, latent TB, with only a small minority (5 to 10%) developing active disease. It is estimated that one third of the human population worldwide may have latent M. tuberculosis infection. Latent TB is characterized by an efficient immune response that contains the infection in a nonpathological and noncontagious state, within a specific, dynamic structure called the granuloma. Interactions between M. tuberculosis and the immune system play a crucial role in determining the outcome of the disease, and are mediated by various pattern recognition receptors (PRRs) expressed in cells of the innate immune system and in nonimmune cells. These interactions may modulate the immune response in favor of the bacillus, by allowing it to persist within host phagocytes. They may also favor the host, by inducing immune defenses, such as autophagy, phagosome maturation, apoptosis and various bactericidal mechanisms. PMID- 21622264 TI - Nitric oxide and thermogenesis--challenge in molecular cell physiology. AB - Only recently we can link thermogenesis, mitochondria, nitric oxide, and redox regulation in biochemical terms. Currently, we are discussing these processes from the aspect of fundamental principles of molecular physiology. Thus, the present article highlights both cell physiology and the principles of the maintenance of energy homeostasis in organisms. Energy homeostasis means much more than simple combustion; adipose tissues at this point of evolution development are related to a broad spectrum of metabolic disturbances and all aspects of cellular remodeling (i.e. structural, metabolic and endocrine changes). Therefore, this paper addresses not only thermogenesis but also energy homeostasis, oxidative phosphorylation and ATP production, proliferation and differentiation of brown adipocytes, their life and death, mitochondriogenesis and angiogenesis. These processes will be united by molecular players of oxidation/reduction reactions, thus creating the principles based on the redox regulation. PMID- 21622265 TI - Role of chemokines in the pathogenesis of endometriosis. AB - Chemokines, proteins that operate within the body's immune system, play numerous roles in menstruation, bacterial infection, implantation of embryos, and the maintenance of early pregnancy. They are also strongly related to the pathogenesis of endometriosis. Several chemokines including interleukin (IL)-8, growth-related oncogene (GRO) alpha, regulated on activation, normal T expressed and secreted (RANTES), and macrophage inflammatory protein (MIP)-1 are reported to be elevated in the peritoneal fluid (PF) of women with endometriosis. Chemokines IL-8 and GRO alpha as well as epithelial cell-derived neutrophil activating protein (ENA)-78, eotaxin, and interferon-inducible protein (IP)-10 might be involved in macrophage activation, inflammatory reaction, and adhesion of endometriotic tissues in the peritoneal cavity, and enhanced angiogenesis in the progression of endometriosis. The chemokines closely related with the pathogenesis of endometriosis form a complex network locally and systemically in women with the disease. Understanding this network is a key to improving our understanding of endometriosis as well as developing new, more effective therapies. PMID- 21622263 TI - Neural stem cell niches: roles for the hyaluronan-based extracellular matrix. AB - Neural stem/progenitor cells capable of differentiating into the neurons and glial cells that populate the mammalian central nervous system (CNS) persist in specific neural stem cell niches that regulate stem cell proliferation, survival and differentiation. There is growing evidence that the extracellular matrix within neural stem cell niches is required for neural stem cell maintenance. Here, we review findings supporting a pivotal role for the glycosaminoglycan hyaluronan (HA) and its transmembrane receptors in neural stem/progenitor cell proliferation, differentiation and maturation. We also outline findings supporting changing roles for HA as cells become committed to distinct lineages in the brain and spinal cord. PMID- 21622266 TI - HRP and prorenin: focus on the (pro)renin receptor and vacuolar H+-ATPase. AB - The function of prorenin, the precursor of renin, remained unknown until the discovery of the (pro)renin receptor ((P)RR). (Pro)renin binding to this receptor allows angiotensin generation and induces signaling. Thus, (P)RR blockade will exert effects beyond angiotensin suppression. Recently, the (P)RR has been identified as an accessory protein of the vacuolar-type H+-ATPase, with important roles in Wnt signaling. In addition, transgenic animals overexpressing prorenin display the consequences of angiotensin generation, whereas transgenic animals overexpressing the (P)RR display an angiotensin-independent phenotype. Finally, both beneficial and deleterious effects have been described following treatment with the (P)RR antagonist 'handle region peptide' (HRP), while a (P)RR knockout in cardiomyocytes is lethal. This review highlights the latest findings in the (P)RR area, focusing on cardiovascular and renal pathology. It critically addresses the possibility that (pro)renin acts as an agonist of this receptor in vivo, and discusses the efficacy of HRP. Conclusions are that convincing evidence for (pro)renin-(P)RR interaction in vivo is currently lacking and, thus, that the concept of HRP exerting beneficial effects by blocking such interaction remains to be proven. PMID- 21622267 TI - Interaction of free radicals, matrix metalloproteinases and caveolin-1 impacts blood-brain barrier permeability. AB - Free radicals play an important role in cerebral ischemia-reperfusion injury. Accumulations of toxic free radicals such as reactive oxygen species (ROS) and reactive nitrogen species (RNS) not only increase the susceptibility of brain tissue to ischemic damage but also trigger numerous molecular cascades, leading to increased blood-brain barrier (BBB) permeability, brain edema, hemorrhage and inflammation, and brain death. Activating matrix metalloproteinases (MMPs) is a key step in BBB disruption. MMPs are proteolytic zinc-containing enzymes responsible for degradation of the extracellular matrix around cerebral blood vessels and neurons. Free radicals can activate MMPs and subsequently induce the degradations of tight junctions (TJs), leading to BBB breakdown in cerebral ischemia-reperfusion injury. Recent studies revealed that caveolin-1, a membrane integral protein located at caveolae, can prevent the degradation of TJ proteins and protect the BBB integrity by inhibiting RNS production and MMPs activity. The interaction of caveolin-1 and RNS forms a positive feedback loop which provides amplified impacts on BBB dysfunction during cerebral ischemia-reperfusion injury. Here, we reviewed the recent progress in the interactions of RNS, caveolin-1 and MMPs. Current evidence indicates that the interactions of RNS, caveolin-1 and MMPs are critical signal pathways in BBB disruption and infarction enlargement during cerebral ischemia-reperfusion injury. PMID- 21622268 TI - The importance of Ca2+/Zn2+ signaling S100 proteins and RAGE in translational medicine. AB - The Receptor for Advanced Glycation Endproducts (RAGE) is a multiligand receptor involved in a large number of human disorders. Identified first as the receptor for the Advanced Glycation Endproducts (AGEs), RAGE has emerged in recent years as a major receptor for many members of the S100 calcium and zinc binding protein family. The interaction with and the signaling triggered by several S100 proteins such as S100B and S100A12 have been studied in details and have shown concentration and cell type dependent signaling cascades. The S100 protein family consists of more than 20 members which present high amino-acid sequence and structural similarities. These small EF-hand calcium binding proteins interact with a large number of protein targets and are almost all been shown to be involved in cancer. In this review we discuss the recent knowledge about the role of S100 proteins and RAGE in human disorders. PMID- 21622269 TI - Tetrahydrobiopterin attenuates superoxide-induced reduction in nitric oxide. AB - NADPH oxidase, a source of superoxide anion (.O2(-)), can be stimulated by oxidized low-density lipoprotein (OxLDL). We examined whether tetrahydrobiopterin (BH4) could reduce OxLDL-induced .O2(-) production by NADPH oxidase, increasing nitric oxide (NO) synthesis. Endothelial cells incubated with OxLDL produced more .O2(-) (35-67%) than untreated cells, with the highest increase 1 hour after OxLDL addition. The elevated .O2(-) production correlated with the translocation of the p47phox subunit of NADPH oxidase from the cytosol to the membrane. Cells exhibited a marked decrease in both BH4 (83 per cent) and NO (54 per cent) in the same hour following exposure to OxLDL. An NADPH oxidase inhibitor, apocynin, or antioxidant, N-acetyl-L-cysteine, substantially attenuated the reduction in both BH4 and NO. The .O2(-) production was increased when cells were pretreated with an inhibitor of BH4 synthesis and decreased following pretreatment with a BH4 precursor, suggesting that NADPH oxidase-induced imbalance of endothelial NO and .O2(-) production can be modulated by BH4 concentrations. BH4 may be critical in combating oxidative stress, restoring proper redox state, and reducing risk for cardiovascular disease including atherosclerosis. PMID- 21622270 TI - Actinobacteria in indoor environments: exposures and respiratory health effects. AB - Actinobacteria are a large group of Gram-positive bacteria common in the environment, especially in the soil. They are morphologically diverse and extremely versatile in their metabolic activities. They produce tens of thousands of secondary metabolites with different biological activities. Exposure to actinobacteria in indoor environments is probably continuous, since they are both common environmental bacteria and human normal flora. However, the occurrence of some species of spore-forming filamentous actinomycetes has been associated with abnormal and health-hazardous situations, such as moisture damage of the building. The measured concentrations of actinobacteria indoors are low. Higher concentrations have been reported during the remediation work of moisture damaged buildings and in agricultural environments. Exposure to high concentrations of actinobacteria can cause allergic alveolitis. Other respiratory disorders have been reported, too and although the measured concentrations are low, the indoor exposure is always a mixture of many different agents, which may have synergistic effects. In vitro and in vivo studies have shown that actinobacteria are very immunoactive and hence, potential causative agents for respiratory and other disorders. PMID- 21622271 TI - Hutchinson-Gilford progeria syndrome, cardiovascular disease and oxidative stress. AB - Hutchinson-Gilford Progeria Syndrome (HGPS), a rare human disease characterized by premature aging, is mainly caused by the abnormal accumulation of progerin, a mutant form of the mammalian nuclear envelope component lamin A. HGPS patients exhibit vascular alterations and die at an average age of 13 years, predominantly from myocardial infarction or stroke. Animal models of HGPS have been a valuable tool in the study of the pathological processes implicated in the origin of this disease and its associated cardiovascular alterations. Some of the molecular mechanisms of HGPS might be relevant to the process of normal aging, since progerin is detected in cells from normal elderly humans. Conversely, processes linked to normal aging, such as the increase in oxidative stress, might be relevant to the pathogenic mechanisms of HGPS. In this review, we discuss recent advances in the understanding of the molecular mechanisms underlying the cardiovascular alterations associated with HGPS, the potential role of oxidative stress, and therapeutic approaches for the treatment of this devastating disease. PMID- 21622272 TI - Biomarkers for optimal requirements of amino acids by animals and humans. AB - Amino acids are building blocks of proteins and key regulators of nutrient metabolism in cells. However, excessive intake of amino acids can be toxic to the body. Therefore, it is important to precisely determine amino acid requirements by organisms. To date, none of the methods is completely satisfactory to generate comprehensive data on amino acid requirements of animals or humans. Because of many influencing factors, amino acid requirements remain a complex and controversial issue in nutrition that warrants further investigations. Benefiting from the rapid advances in the emerging omics technologies and bioinformatics, biomarker discovery shows great potential in obtaining in-depth understanding of regulatory networks in protein metabolism. This review summarizes the current approaches to assess amino acid requirements of animals and humans, as well as the recent development of biomarkers as potentially functional parameters for recommending requirements of individual amino acids in health and disease. Identification of biomarkers in plasma or serum, which is a noninvasive approach, holds great promise in rapidly advancing the field of protein nutrition. PMID- 21622273 TI - Electrochemical and optical biosensors based on nanomaterials and nanostructures: a review. AB - Nanomaterials and nanostructures exhibit unique size-tunable and shape-dependent physicochemical properties that are different from those of bulk materials. Advances of nanomaterials and nanostructures open a new door to develop various novel biosensors. The present work has reviewed the recent progress in electrochemical, surface plasmon resonance (SPR), surface-enhanced Raman scattering (SERS) and fluorescent biosensors based on nanomaterials and nanostructures. An emphasis is put on the research that demonstrates how the performance of biosensors such as the limit of detection, sensitivity and selectivity is improved by the use of nanomaterials and nanostructures. PMID- 21622274 TI - Dietary flavonoids as cancer-preventive and therapeutic biofactors. AB - Flavonoids are present in many plants, and hence, in foods and ingredients derived from them. These polyphenolic compounds have attracted renewed attention as potential anticarcinogens, and the molecular mechanisms of their anticarcinogenic effects and their bioavailability have been extensively explored. In this review, we focus on the major dietary flavonoids; flavones, flavonols, and flavan-3-ols (catechins), and evaluate their roles in cancer prevention. After absorption with or without metabolic conjugation, flavonoids are transported to target organs where they exert their anticarcinogenic activity. The molecular mechanisms of the anticarcinogenic effects of flavonoids include their antagonistic effect on the aryl hydrocarbon receptor (AhR), and regulation of phase I and II drug metabolizing enzymes and phase III transporters. Experimental evidence suggests that flavonoids modulate signal transduction pathways at each stage of carcinogenesis. The interactions between flavonoids and biomolecules in vivo must be investigated in detail to identify specific targets. In addition, the potential side effects should be considered when flavonoid supplements are used for cancer prevention. Therefore, the use of flavonoids as chemopreventive agents should be further investigated to establish safe levels of flavonoid intake. PMID- 21622275 TI - Advances in homeopathy and immunology: a review of clinical research. AB - The present paper reviews the clinical research carried out over the past three decades to evaluate the effectiveness of homeopathy for the treatment of respiratory allergies, common upper respiratory tract infections, otorhinolaryngologic complaints, and rheumatic diseases. We include in the analysis both randomised and non-randomised trials, assigning them different weightings in the final balance of evidence, on the basis of semi-quantitative criteria. Overall, the literature concerning a total of 83 original studies suggests that homeopathy may have significant effects in some conditions, e.g. Galphimia glauca (low homeopathic dilutions/dynamizations) in allergic oculorhinitis, Anas barbariae (high homeopathic dilution/dynamization) in influenza-like syndromes, classical individualised homeopathy in otitis, in allergic complaints and in fibromyalgia, and a few low-potency homeopathic complexes in sinusitis, rhinoconjunctivitis, arthritis. The evidence for individualised homeopathic therapy in the field of upper respiratory tract infections and for homeopathic immunotherapy in respiratory allergies is more conflicting. Pragmatic equivalence trials suggest that, in primary care, homeopathic treatment is not inferior to conventional treatment. A larger number of observational studies and of clinical trials -- conducted in a methodologically correct manner without altering the treatment setting-- are needed before sure conclusions concerning the application of homeopathy for specific diseases can be drawn. PMID- 21622276 TI - Origin, maturation and recruitment of mast cell precursors. AB - Mast cells have gained increased recognition as immunomodulators playing a role in a variety of physiological and pathological processes. They were first described in 1879, but their origin remained controversial for almost a century. Today, it is known that mast cells are present in the bone marrow as committed mast cell precursors. They leave the bone marrow as progenitors and complete their maturation at peripheral sites. Investigations on the maturation of bone marrow derived mast cells focused on bone marrow cultured in the presence of interleukin-3 (IL-3) and stem cell factor (SCF). SCF is essential for mast cell survival and mice that lack either SCF or the receptor for SCF are mast cell deficient. It is the microenvironment surrounding the mast cell that determines its mature phenotype. SCF, IL-3 and IL-9 have been identified among the most important cytokines for regulation of mast cell growth and differentiation. Several factors have been identified as chemoattractants for mast cells, but their exact mechanism of action remains unclear. Mast cell recruitment is most likely a combination of the direct effect of mast cell mediators on the mast cell progenitor as well as the indirect effect of these mediators on other cell types. PMID- 21622277 TI - Neuroinflammation and cell therapy for Parkinson's disease. AB - Cell therapy is a promising therapeutic alternative for Parkinson's disease, and one possible limiting factor may be that the pathological environment of PD is hostile for the process of neurogenesis, including grafted stem cells survival, proliferation, migration and dopaminergic neuronal fate specification along with maturation of the immature neurons and ultimately integration of the new neuronal progeny into functional neuronal circuits. Uncontrolled microglial activation and neuroinflammation contributes to neuronal damage in PD. Similarly, the microglia derived inflammatory mediators may also influence grafted stem cells. Thus, we discuss reactive microgliosis and sustained, chronic neuroinflammation in PD, together with cytokine-dependent neurotoxicity and inflammation-derived oxidative stress on dopaminergic neuron in the substantia nigra pars compacta substantia nigra pars compacta (SNpc). Based on these, we further summarize the interaction between neuroinflammation and stem cells, and conclude that neuroinflammation acts as double-edged swords, instead of simply beneficial or detrimental, and stem cells display immunomodulatory functions beneficial for dopaminergic neurons via an anti-inflammatory action in PD. PMID- 21622278 TI - Cancer immunology and colorectal cancer recurrence. AB - The recurrence of a cancer - local or distant (metastasis) - is manifested by the persistence of cancer cells in the organism after the ablation of the primary lesion, an ineffective anticancer immune response, and by the activity of biological/immunological factors that can stimulate and sustain its development. This review focuses on colorectal carcinoma and discusses some aspects of cancer immunology regarding cancer development and its recurrence. It is addressed also to the clinician to provide new insights helpful for designing better therapeutic strategies and patient's follow up. Therapeutic approaches used during and after surgical treatments, found capable of modulating immunity (differently affecting disease outcome), will also be described. PMID- 21622279 TI - Low protein diets are mainstay for management of chronic kidney disease. AB - Low protein diets, made either of natural foods or of L-essential amino acids and/or their nitrogen-free ketoanalogues, are feasible, safe, and efficient means to reduce disease progression in patients with chronic kidney disease and do not prejudice patient outcomes once they get into Renal Replacement Therapy. They ameliorate symptomatology, grant a positive nitrogen balance, reduce proteinuria, improve osteodystrophy and lipid profile, reduce serum concentrations of uric acid, phosphate, and maintain plasma bicarbonate within normal limits thus preventing metabolic acidosis. They also reduce the number of hypotensive drugs and the quantity of erythropoietin to be administered to achieve target hemoglobin concentrations, and do not deteriorate quality of life. On the contrary, they retard progression of chronic kidney disease. There is a need to motivate patients to increase adherence to. PMID- 21622280 TI - P2 receptors and extracellular ATP: a novel homeostatic pathway in inflammation. AB - Inflammation is an important homeostatic response, which is managed by a complex network of interrelated pathways that determine the level, intensity and localization of inflammation. We now know that purinergic signalling is one of the pathways influencing the initiation, progression and down-modulation of the inflammatory response. Here, we review recent evidence on the role in inflammation of the purinergic signalling system, which is comprised of extracellular ATP, P2 receptors and ecto-enzyme cascades. Recent animal studies with a newly developed bioluminescent ATP probe (pmeLUC), enabling measurement of pericellular ATP in situ, have provided proof that ATP is present in inflamed tissues in vivo at extracellular concentrations sufficient for P2 receptor activation. Increased extracellular ATP levels amplify inflammation in vivo by promoting leukocyte recruitment and NALP3-inflammasome activation via P2X7. Lowering extracellular ATP levels in inflamed tissues, for instance by stimulating its breakdown, inhibits the inflammatory response in vivo. In view of its important role in inflammation, the purinergic signalling system is bound to yield novel therapeutic opportunities for the treatment of inflammatory diseases. PMID- 21622281 TI - Alternatively spliced tissue factor pathway inhibitor: functional implications. AB - Tissue factor pathway inhibitor (TFPI) is a factor Xa dependent inhibitor of tissue factor initiated blood coagulation. In recent years several alternatively spliced forms of TFPI have been identified. These alternatively spliced forms have different C-terminal regions and have different mechanisms for association with cell surfaces. They are differentially expressed in human and mouse tissues and may have distinct physiological functions. PMID- 21622282 TI - Genetic susceptibility to lung cancer. AB - Lung cancer is a highly environmental disease, but cancer researchers have long been interested in investigating genetic susceptibility to lung cancer. This paper is a historical review and provides updated perspectives on lung cancer susceptibility research. The recent introduction of easier genotyping methods and the availability of an almost complete human genome database facilitated the association study to thousands of cases and controls for millions of genetic markers. Discoveries in the field of behavior genetics, that is, the genetic aspects of smoking behavior and nicotine addiction, unexpectedly indicated that polymorphisms in the human central nervous system play an important role in eventually leading to lung cancer. These findings were achieved by using comprehensive approaches, such as a genome, transcriptome, or proteome approach, and the studies were often conducted without a hypothesis. Another-omics approach, the "adductome" or "exposome" approach to how life style information can be integrated into the framework of genetic association studies, has recently emerged. These new paradigms will influence the area of lung cancer risk evaluation in genome cohort studies. PMID- 21622283 TI - Possible involvement of the (pro)renin receptor-dependent system in the development of insulin resistance. AB - It is widely acknowledged that activation of the renin-angiotensin system impairs insulin sensitivity. Pharmacological inhibition of the (pro)renin receptor dependent system has shown beneficial effects in diabetic nephropathy, retinopathy and hypertensive cardiac damage in animal models. Previously, we showed that fructose feeding stimulated nonproteolytic activation of prorenin and subsequent production of angiotensin II in skeletal muscle in rats, and that inhibition of the (pro)renin receptor-dependent system improved the development of fructose feeding-induced insulin resistance. In addition, our current preliminary study suggests that local angiotensin II generation in skeletal muscle and adipose tissues induced by nonproteolytic activation of prorenin is involved in the development of spontaneous insulin resistance in type 2 diabetic rats. In this review, we will briefly summarize the possible contribution of the (pro)renin receptor-dependent system to the pathogenesis of insulin resistance, with a focus on how the nonproteolytic activation of prorenin contributes to the development of insulin resistance. PMID- 21622284 TI - Cancer stem cells: perspectives of new therapeutical approaches for breast cancer. AB - Currently stem cells are hypothesized to play a central role in the origin, spread and resistance to treatment of breast cancer. Common anticancer therapy is effective but transient, with tumor relapse and metastatic disease often occurring. For therapy to be more effective, debulking of differentiated tumors must occur followed by targeting of the remaining surviving often quiescent tumor stem cells. New therapeutics aimed at cancer stem cells are achieved through non immunological and immunological methods. The former include elective ABC drug transporters or the heat shock protein 90 inhibition, targeting the self-renewal signalling pathways or the EMT program, differentiation therapy, or other interventions to eliminate BrCSCs. The latter include targeting specific antigens expressed on BrCSCs, dendritic cells (DCs) based vaccination and blockers of the extrinsic signals at CSC niche. Here all these novel approaches related to breast cancer stem cells are described. PMID- 21622285 TI - Tissue factor signaling: a multi-faceted function in biological processes. AB - Tissue factor (TF), originally discovered to initiate coagulation, is more recently recognized to be involved in other biological processes, such as migration and anti-apoptosis. TF-mediated signaling regulates gene expression and protein synthesis, leading to alterations in cellular behavior. The proteolytic activity of factor VIIa (FVIIa), beta-1 integrin interaction and protease activated receptor (PAR) activation are some of the key events involved in TF signaling. Post-translational modifications of TF may regulate signaling but this remains elusive. In vivo studies have established that TF signaling severely contributes to processes like angiogenesis, cancer growth and inflammation. This review focuses on the mechanism underlying TF-mediated intracellular signaling with its related physiological and mainly pathological consequences. PMID- 21622286 TI - Differential expression profiles of mRNAs, miRNAs and proteins during embryo implantation. AB - Embryo implantation is a critical step in mammalian reproduction and includes apposition of the blastocyst, attachment to the uterine epithelium and decidualization of the uterine stroma. However, the key mechanism underlying embryo implantation remains to be elucidated. In recent years, high-throughput transcriptomic and proteomic approaches make it possible for studying the expression levels of thousands of genes and proteins simultaneously and thus accelerate the understanding of gene regulatory networks involved in embryo implantation. This review briefly summarizes the advance on mRNA, miRNA and protein expression profile analysis during embryo implantation in human and a variety of animal models. PMID- 21622288 TI - Mother knows best: lessons from fetomaternal tolerance applied to cancer immunity. AB - Failure of the immune system to recognize and eradicate tumor cells has deadly consequences. It is possible that the normal host response to the inflammatory environment created by many cancers - the body's natural attempt at wound repair and restoration of tissue integrity - is one of counter-regulation that paradoxically favors tumor growth. A physiologic condition where this situation is favorable (and even required) is that of normal pregnancy, where blastocyst implantation creates endometrial inflammation, and the maternal response in turn supports angiogenesis and tolerance required for placentation. Lack of such inflammation and resultant maternal immunologic engagement can lead to serious pregnancy complications including fetal loss, highlighting how important the fetomaternal immunologic dialogue is for survival. Here, we describe how the dynamics of fetomaternal tolerance can help disentangle complex cancer/host immunologic interactions and provide new avenues for immunologic reconstitution in patients with cancer. PMID- 21622287 TI - Immunologic mapping of glycomes: implications for cancer diagnosis and therapy. AB - Cancer associated glycoconjugates are important biomarkers, as exemplified by globo-H, CA125, CA15.3 and CA27.29. However, the exact chemical structures of many such biomarkers remain unknown because of technological limitations. In this article, we propose the "immunologic mapping" of cancer glycomes based on specific immune recognition of glycan structures, which can be hypothesized theoretically, produced chemically, and examined biologically by immuno-assays. Immunologic mapping of glycans not only provides a unique perspective on cancer glycomes, but also may lead to the invention of powerful reagents for diagnosis and therapy. PMID- 21622289 TI - Effect of sulfated glycosaminoglycans on tumor invasion and metastasis. AB - Metastasis is the most devastating aspect of the tumor, being the main cause of morbidity and mortality in cancer patients. The events that lead to tumor invasion and metastasis depend fundamentally on the initial aquisition of a mesenchymal phenotype by the primary carcinoma, which provides the necessary machinary for invasion, intravasation, vascular transport, extravasation and tumor colonization. These events are orquestrated by different growth factors, proteoglycans and adhesion molecules, acting at the surface of cells. The anticoagulant heparin binds several of these molecules and can regulate the interactions that occur during tumor invasion and metastasis. For example, heparin modulates the binding of FGF-2 to its tyrosine kinase receptor during tumor proliferation, and the binding of growth factors involved in epithelial to mesenchymal transition during tumor invasion. It also binds P-selectin on activated platelets, preventing tumor cell-platelet interaction during hematogeneous metastasis. In this review, we discuss the role of sulfated glycosaminoglycans during tumor invasion and metastasis, and the possible therapeutic use of heparin analogs on cancer treatment. PMID- 21622290 TI - Outdoor thermal comfort. AB - A review of the various approaches in understanding outdoor thermal comfort is presented. The emphasis on field surveys from around the world, particularly across Europe, enables us to understand thermal perception and evaluate outdoor thermal comfort conditions. The consistent low correlations between objective microclimatic variables, subjective thermal sensation and comfort outdoors, internationally, suggest that thermophysiology alone does not adequate describe these relationships. Focusing on the concept of adaptation, it tries to explain how this influences outdoor comfort, enabling us to inhabit and get satisfaction from outdoor spaces throughout the year. Beyond acclimatization and behavioral adaptation, through adjustments in clothing and changes to the metabolic heat, psychological adaptation plays a critical role to ensure thermal comfort and satisfaction with the outdoor environment. Such parameters include recent experiences and expectations; personal choice and perceived control, more important than whether that control is actually exercised; and the need for positive environmental stimulation suggesting that thermal neutrality is not a pre-requisite for thermal comfort. Ultimately, enhancing environmental diversity can influence thermal perception and experience of open spaces. PMID- 21622292 TI - Parent satisfaction with the electronic medical record in an academic pediatric rheumatology practice. AB - BACKGROUND: Patient satisfaction has not been widely studied with respect to implementation of the electronic medical record (EMR). There are few reports of the impact of the EMR in pediatrics. OBJECTIVE: The objective of this study was to assess the impact of implementation of an electronic medical record system on families in an academic pediatric rheumatology practice. METHODS: Families were surveyed 1 month pre-EMR implementation and 3 months post-EMR implementation. RESULTS: Overall, EMR was well received by families. Compared with the paper chart, parents agreed the EMR improved the quality of doctor care (55% or 59/107 vs 26% or 26/99, P < .001). More parents indicated they would prefer their pediatric physicians to use an EMR (68% or 73/107 vs 51% or 50/99, P = .01). CONCLUSIONS: Transitioning an academic pediatric rheumatology practice to an EMR can increase family satisfaction with the office visit. PMID- 21622291 TI - Chemokines: key players in cancer progression and metastasis. AB - Instructed cell migration is a fundamental component of various biological systems and is critical to the pathogenesis of many diseases including cancer. Role of chemokines in providing navigational cues to migrating cancer cells bearing specific receptors is well established. However, functional mechanisms of chemokine are not well implicit, which is crucial for designing new therapeutics to control tumor growth and metastasis. Multiple functions and mode of actions have been advocated for chemokines and their receptors in the progression of primary and secondary tumors. In this review, we have discussed current advances in understanding the role of the chemokines and their corresponding receptor in tumor progression and metastasis. PMID- 21622293 TI - Tanshinone IIA-induced attenuation of lung injury in endotoxemic mice is associated with reduction of hypoxia-inducible factor 1alpha expression. AB - Inhibiting hypoxia-inducible factor (HIF)-1alpha activity has been proposed as a novel therapeutic target in LPS-induced sepsis syndrome. We have reported that tanshinone IIA (TIIA) can reduce LPS-induced lethality and lung injury in mice, but the precise mechanisms have not been fully described. Therefore, the present study investigated whether the protective effect of TIIA was related to the inhibition of LPS-induced HIF-1alpha expression and what mechanisms accounted for it. This study showed that TIIA pretreatment improved LPS-induced biochemical and cellular changes and reduced the production of inflammatory cytokines. Pretreatment with TIIA decreased LPS-induced HIF-1alpha expression in vivo and in vitro. TIIA did not affect the LPS-induced HIF-1alpha mRNA level but inhibited HIF-1alpha protein translation by the inhibition of the PI3K/AKT and MAPK pathways and related protein translational regulators, such as p70S6K1, S6 ribosomal protein, 4E-BP1, and eIF4E, and promoted HIF-1alpha protein degradation via the proteasomal pathway in LPS-stimulated macrophages. These observations partially explain the antiinflammatory effects of TIIA, which provides scientific basis for its application for the treatment of acute lung injury/acute respiratory distress syndrome or sepsis. PMID- 21622294 TI - Symposium on scientific literacy: Introduction. PMID- 21622295 TI - Botanical literacy: What and how should students learn about plants? AB - Botanists benefit from a scientifically literate society and an interested and botanically literate student population, and we have opportunities to promote literacy in our classes. Unfortunately, scientific illiteracy exists, in part, because students are technologically advanced but lack intellectual curiosity and rigor. Botanical illiteracy results from several interacting factors, including a lack of interest in plants and infrequent exposure to plant science before students reach college. If scientific or botanical literacy is a goal, we must understand what literacy means and how we can help students reach that goal. A model of biological literacy recognizes four levels; students enter courses at the lowest level possessing misconceptions about concepts; however, misconceptions can be used to our advantage, especially by using concept inventories. Inquiry-based instruction is advocated for all science courses, and learning theory supports inquiry. Seven principles of learning inform recommendations about how botanists should teach, including using themes and "thinking botanically" to illustrate all biological concepts. Overall, consideration of the botanical content taught is less critical than the methods used to teach that content. If botanists emphasize thinking and process skills with an understanding of concepts, we will prepare scientifically literate students and citizens and benefit from our efforts. PMID- 21622296 TI - Improving the public understanding of science: New initiatives. AB - The United States may be on the brink of losing its global edge in science. Many American students are underprepared for and uninterested in the scientific and technical careers they may be asked to take on. Furthermore, these students, their teachers, and the broader public lack basic understandings of what science is and how it works, which may negatively impact their ability to make reasoned and informed decisions about science-related issues. We describe two unique and recently developed projects designed to help tackle these problems by improving public understanding of and interest in science. The Coalition on the Public Understanding of Science is a grassroots effort to lower the barriers between the scientific community and the public. It aims to inspire broad appreciation of science, inform the public about the nature and process of science, and make science accessible to everyone. Understanding Science is a web-based project that aims to improve teacher understanding of the nature of the scientific enterprise, to provide resources that encourage and enable kindergarten through undergraduate (K-16) teachers to reinforce the nature of science throughout their teaching, and to serve as a clear and accessible reference that accurately portrays the scientific endeavor. The botanical and broader scientific communities are invited to participate in these efforts. PMID- 21622297 TI - What's next for science communication? Promising directions and lingering distractions. AB - In this essay, we review research from the social sciences on how the public makes sense of and participates in societal decisions about science and technology. We specifically highlight the role of the media and public communication in this process, challenging the still dominant assumption that science literacy is both the problem and the solution to societal conflicts. After reviewing the cases of evolution, climate change, food biotechnology, and nanotechnology, we offer a set of detailed recommendations for improved public engagement efforts on the part of scientists and their organizations. We emphasize the need for science communication initiatives that are guided by careful formative research; that span a diversity of media platforms and audiences; and that facilitate conversations with the public that recognize, respect, and incorporate differences in knowledge, values, perspectives, and goals. PMID- 21622298 TI - Long-term relationships among atmospheric CO2, stomata, and intrinsic water use efficiency in individual trees. AB - Leaf-level responses to increases in atmospheric carbon dioxide (CO(2)) concentrations could have large implications for water and carbon cycles. We investigated whether stomatal density, guard cell length, and intrinsic water use efficiency (iWUE) of 27 individual trees growing at the Arnold Arboretum in Boston, Massachusetts have responded to changing environmental conditions over the last 100 years. We examined leaves from 74 herbarium specimens collected from three genera-Acer (maples), Quercus (oaks), and Carpinus (hornbeams)-from 1893 to 2006. During this period, global average atmospheric CO(2) concentrations increased by approximately 29% (86 ppm), and temperatures in Boston increased by 1.8 degrees C. Stomatal density and guard cell length were negatively correlated in oaks and hornbeams. Although stomatal density declined and guard cell length increased over time, the changes were not dependent on the magnitude of changes in CO(2) concentrations. Intrinsic WUE did not change significantly over time. Our findings suggest that iWUE may not respond to changes in CO(2) concentrations over the lifetimes of individual trees, possibly because of compensating changes in stomatal density and guard cell size. We provide an example of a method that can enable researchers to differentiate between genetic and plastic responses to global change in long-lived trees. PMID- 21622299 TI - Wollemia nobilis (Araucariaceae): Branching, vasculature and histology in juvenile stages. AB - We present a preliminary description of the morphology and anatomy of contrasted axes in the recently discovered conifer Wollemia nobilis, based on clonally propagated material. The novelty of this discovery and the tree's size and rarity has led to global interest and a comprehensive and successful conservation program. Our results should serve as a model on which future studies of this tree and other members of the Araucariaceae can be based. The specimens studied are mimics of the architecture of the mature tree, with precise distinction between orthotropic (= trunk) axes, which have radial symmetry (spiral phyllotaxy) and plagiotropic (= branch) axes with dorsiventral symmetry (as a modified decussate phyllotaxy). Trunk axes develop irregular pseudowhorls of branches that originate within the terminal bud by syllepsis, their extension coincident with that of the parent axis. The two kinds of axes show considerable anatomical differences, but are still based on the common feature of a single stelar-derived trace to each leaf that becomes subdivided in the cortex, a feature of the whole family. Trunk axes include extended cortical leaf and branch traces associated with abundant sclerenchyma, but branches have short cortical leaf traces, no branch traces, and limited sclerenchyma. Reiteration is limited and largely involves the formation of basal suckers on the trunk. Branches normally remain unbranched, but can do so most often when damaged. This study thus emphasizes the phenomenon of axis differentiation in conifers, which has been little investigated anatomically, but could be very important in the identification of fossils. PMID- 21622300 TI - A cellular study of teosinte Zea mays subsp. parviglumis (Poaceae) caryopsis development showing several processes conserved in maize. AB - The evolutionary history of maize (Zea mays subsp. mays) is of general interest because of its economic and scientific importance. Here we show that many cellular traits described previously in developing caryopses of maize are also seen in its wild progenitor teosinte (Zea mays subsp. parviglumis). These features, each with a possible role in development, include (1) an early programmed cell death in the maternal placento-chalazal (P-C) layer that may lead to increased hydrolytic conductance to the developing seed; (2) accumulation of phenolics and flavonoids in the P-C layer that may be related to antimicrobial activity; (3) formation of wall ingrowths in the basal endosperm transfer layer (BETL); (4) localization of cell wall invertase in the BETL, which is attributed to the increased transport capacity of photosynthates to the sink; and (5) endoreduplication in endosperm nuclei suggested to contribute to increased gene expression and greater sink capacity of the developing seed. In maize caryopsis, these cellular traits have been previously attributed to domestication and selection for larger seed size and vigor. Given the conservation of the entire cellular program in developing teosinte caryopses described here, we suggest that these traits evolved independently of domestication and predate human selection pressure. PMID- 21622301 TI - Variation in flowering size and age of a facultative biennial, Aster kantoensis (Compositae), in response to nutrient availability. AB - Although the flowering of facultative biennials is size-dependent, flowering size varies markedly within a single population as well as among populations. In this study, 15 half-sib families of the facultative biennial Aster kantoensis were grown from seeds at three nutrient levels (low, medium, and high). A significant nutrient * family interaction effect was found for bolting size, and among-family variation in bolting size increased with decreasing nutrient level. Growth from bolting to flowering tended to be greatest at the high nutrient level. Such responses of bolting size and growth from bolting to flowering resulted in an increase in flowering size at the high nutrient level and a significant variation in its reaction norm among families. For flowering age, there was a significant interaction of nutrient * family, and its among-family variation increased with decreasing nutrient levels, as was the case with bolting size. These results indicate that genetic variation in phenotypic plasticity of bolting size with nutrient availability was one cause of the variation in flowering size and age in the A. kantoensis population on the floodplain with the spatially heterogeneous nutrient availability. Moreover, responses of growth from bolting to flowering to nutrient availability could enhance the variation in flowering size. PMID- 21622302 TI - Complex interactions between a legume and two grasses in a subalpine meadow. AB - Interactions between plants are a complex combination of positive and negative interactions, with the net outcome depending on environmental contexts. The more frequent association of Trifolium alpinum (legume) with Festuca eskia than with Nardus stricta (grasses) in many Pyrenean subalpine meadows suggests a differential ability to use nitrogen (N) derived from N(2) fixation. In the field, we investigated the interactions between the legume and grasses and, in the glasshouse, the transfer of (15)N from the legume to the grasses. In one grass-Trifolium mixture, the legume had a strong positive effect on the biomass and N content of the grass as compared to pure grass stands. When both grasses grew together with the legume, only Festuca benefited from the presence of Trifolium but, surprisingly, the benefit decreased with increasing Trifolium abundance. Leaf labeling experiments with (15)N-NH(4)(+) revealed a higher transfer of (15)N from Trifolium to Festuca than to Nardus, suggesting a more direct N pathway between the two species. This more direct pathway could prevent Nardus from benefiting from the legume N in the three-species mixtures. Thus, the positive interactions between N-fixers and nonfixers appear to be largely species specific and to depend strongly on the species in the plant assemblage. PMID- 21622303 TI - Variation in the impact of climate change on flowering phenology and abundance: An examination of two pairs of closely related wildflower species. AB - Variability in plant phenological responses to climate change is likely to lead to changes in many ecological relationships as the climate continues to change. We used a 34-yr record of flowering times and flower abundance for four species (two Delphinium [Ranunculaceae] species and two Mertensia [Boraginaceae] species) from a subalpine plant community near the Rocky Mountain Biological Laboratory to test the hypothesis that the phenologies of early-flowering species change more rapidly in response to climatological and other abiotic cues than do late flowering species, a pattern previously found in plant communities in North America and Europe. We also explored a related hypothesis, that flower abundance of late-flowering species is more responsive to changes in climate than that of early-flowering species. The Delphinium species did not support these hypotheses, but the Mertensia species did. The difference between the peak flowering times of the early and late Mertensia species is expanding, leading to a period of diminished resources for pollinators that specialize on this genus. Mertensia ciliata populations are already severely declining in our study area, possibly as a result of earlier snowmelt. Together, these results show that the reported differences between early- and late-flowering species may be widespread, but they are not ubiquitous. PMID- 21622304 TI - Differential expression of candidate salt-tolerance genes in the halophyte Helianthus paradoxus and its glycophyte progenitors H. annuus and H. petiolaris (Asteraceae). AB - Adaptation to different habitats is considered to be a major force in the generation of organismal diversity. Understanding the genetic mechanisms that produce such adaptations will provide insights into long-standing questions in evolutionary biology and, at the same time, improve predictions of plant responses to changing environmental conditions. Here we used semiquantitative RT PCR to study the expression of eight candidate salt-tolerance genes in leaves of the highly salt-tolerant diploid hybrid species Helianthus paradoxus and its salt sensitive progenitor species H. annuus and H. petiolaris. Samples were collected after germination and growth under four different treatments: nonsaline (control), near-natural saline, saline with increased K(+), and saline with decreased Mg(2+) and Ca(2+). Three individuals from three populations per species were used. The hybrid species H. paradoxus constitutively under- or overexpressed genes related to potassium and calcium transport (homologues of KT1, KT2, ECA1), suggesting that these genes may contribute to the adaptation of H. paradoxus to salinity. In two other genes, variation between populations within species exceeded species level variation. Furthermore, homologues of the potassium transporter HAK8 and of a transcriptional regulator were generally overexpressed in saline treatments, suggesting that these genes are involved in sustained growth under saline conditions in Helianthus. PMID- 21622305 TI - Origins of domestication and polyploidy in oca (Oxalis tuberosa; Oxalidaceae). 3. AFLP data of oca and four wild, tuber-bearing taxa. AB - Many crops are polyploids, and it can be challenging to untangle the often complicated history of their origins of domestication and origins of polyploidy. To complement other studies of the origins of polyploidy of the octoploid tuber crop oca (Oxalis tuberosa) that used DNA sequence data and phylogenetic methods, we here compared AFLP data for oca with four wild, tuber-bearing Oxalis taxa found in different regions of the central Andes. Results confirmed the divergence of two use-categories of cultivated oca that indigenous farmers use for different purposes, suggesting the possibility that they might have had separate origins of domestication. Despite previous results with nuclear-encoded, chloroplast expressed glutamine synthetase suggesting that O. picchensis might be a progenitor of oca, AFLP data of this species, as well as different populations of wild, tuber-bearing Oxalis found in Lima Department, Peru, were relatively divergent from O. tuberosa. Results from all analytical methods suggested that the unnamed wild, tuber-bearing Oxalis found in Bolivia and O. chicligastensis in NW Argentina are the best candidates as the genome donors for polyploid O. tuberosa, but the results were somewhat equivocal about which of these two taxa is the more strongly supported as oca's progenitor. PMID- 21622306 TI - Spore wall ultrastructure in the early lycopsid Leclercqia (Protolepidodendrales) from the Lower Devonian of North America: Evidence for a fundamental division in the lycopsids. AB - Documenting the morphology and ultrastructure of spores from known Silurian Devonian plants clarifies organization and probable affinities of dispersed spores and contributes to analyses of evolutionary changes and phylogenetic relationships in early plants. In this study of fossil in situ spores from the early protolepidodendralean lycopsid Leclercqia, we identified new characters including an additional synapomorphy of the ligulate lycopsid clade. A detailed light (LM), scanning electron (SEM), and transmission electron microscope (TEM) analysis of spores from two species of Leclercqia from the Lower Devonian (Emsian) of New Brunswick, eastern Canada, L. andrewsii and L. complexa, shows both are homosporous, yielding spores belonging to the dispersed spore form taxon Acinosporites lindlarensis. Important features of wall ultrastructure include the presence of a paraexospore, peculiar exospore-derived, peg-like structures located in the gap between the outer exospore/inner paraexospore, and multilamellate regions in the interradial areas of the proximal surface. Similar interradial multilamellate regions occur in other ligulate lycopsids (fossil and extant). This character is probably a further synapomorphy for the ligulate lycopsid clade, within which heterosporous lycopods form a monophyletic group. These data suggest the ligule and interradial multilamellate region appeared prior to heterospory. PMID- 21622307 TI - Nondestructive estimation of anthocyanins and chlorophylls in anthocyanic leaves. AB - The anthocyanin and chlorophyll contents in leaves provide valuable information about the physiological status of plants. Thus, there is a need for accurate, efficient, and practical methodologies to estimate these biochemical parameters of vegetation. In this study, we tested the performance and accuracy of several nondestructive, reflectance-based techniques for estimating anthocyanin and chlorophyll contents in leaves of four unrelated species, European hazel (Corylus avellana), Siberian dogwood (Cornus alba =Swida alba), Norway maple (Acer platanoides), and Virginia creeper (Parthenocissus quinquefolia), with widely variable pigment content and composition. An anthocyanin reflectance index, which uses reflectances in the green and red edge spectral bands, and a modified anthocyanin reflectance index, employing, in addition, the near-infrared (NIR) band, were able to accurately estimate leaf anthocyanin for all species taken together with no reparameterization of algorithms. Total chlorophyll content was accurately estimated by a red edge chlorophyll index that uses spectral bands in the red edge and the NIR. These approaches can be used to estimate anthocyanin and chlorophyll nondestructively and allow the development of simple handheld field instrumentation. PMID- 21622308 TI - A weed-crop complex in sorghum: The dynamics of genetic diversity in a traditional farming system. AB - Despite the major ecological and economic impacts of gene flow between domesticated plants and their wild relatives, many aspects of the process, particularly the relative roles of natural and human selection in facilitating or constraining gene flow, are still poorly understood. We developed a multidisciplinary approach, involving both biologists and social scientists, to investigate the dynamics of genetic diversity of a sorghum weed-crop complex in a village of Duupa farmers in northern Cameroon. Farmers distinguish a gradient from weedy morphotypes (naa baa see, haariya, and genkiya) to domesticated morphotypes; haariya and genkiya have intermediate morphological traits. We investigated the pattern of diversity in this complex using both morphological and genetic data. Our biological results are interpreted in the light of data on farmers' taxonomy and practices such as spatial pattern of planting and plant selection. Both morphological and genetic data are congruent with farmers' taxonomy and confirm the introgressed status of intermediate weedy morphotypes. Farmers actively select against weedy morphotypes, but several practices unconsciously favor gene flow. Furthermore, haariya and genkiya may facilitate introgression between naa baa see and domesticated morphotypes by virtue of their intermediate flowering period and their mode of management by farmers. PMID- 21622309 TI - Genetic variation in the allotetraploid Dryopteris corleyi (Dryopteridaceae) and its diploid parental species in the Iberian Peninsula. AB - Studies on genetic diversity help us to unveil the evolutionary processes of species and populations and can explain several traits of diploid-polyploid complexes such as their distributions, their breeding systems, and the origin of polyploids. We examined the allozyme variation of Dryopteris aemula and D. oreades, diploid ferns with highly fragmented habitats, and the allotetraploid D. corleyi to (1) analyze the putative relationship between both diploids and the tetraploid, (2) compare the levels of genetic variation among species and determine their causes, and (3) assess the breeding system of these taxa. The allozymic pattern of D. corleyi confirms that it derived from D. aemula and D. oreades. The lack of genetic diversity in D. aemula, a species of lowland habitats, may be due to genetic drift associated with the contraction of populations in the last glaciation. By contrast, the alpine D. oreades had moderate intrapopulation genetic variation, which may derive from the expansion of populations during the last glaciation. In the latter species, low interpopulational variation suggested effective gene flow (spore exchange), and genotype frequencies in Hardy-Weinberg equilibrium indicated cross-fertilization of gametophytes. Evolutionary history appears to be an essential element in the interpretation of genetic variation of highly fragmented populations. PMID- 21622310 TI - Specialization clines in the pollination systems of agaves (Agavaceae) and columnar cacti (Cactaceae): A phylogenetically controlled meta-analysis. AB - The biogeography of plant-animal interactions is a novel topic on which many disciplines converge (e.g., reproductive biology, biogeography, and evolutionary biology). Narrative reviews have indicated that tropical columnar cacti and agaves have highly specialized pollination systems, while extratropical species have generalized systems. However, this dichotomy has never been quantitatively tested. We tested this hypothesis using traditional and phylogenetically informed meta-analysis. Three effect sizes were estimated from the literature: diurnal, nocturnal, and hand cross-pollination (an indicator of pollen limitation). Columnar cactus pollination systems ranged from purely bat-pollinated in the tropics to generalized pollination, with diurnal visitors as effective as nocturnal visitors in extratropical regions; even when phylogenetic relatedness among species is taken into account. Metaregressions identified a latitudinal increase in pollen limitation in columnar cacti, but this increase was not significant after correcting for phylogeny. The currently available data for agaves do not support any latitudinal trend. Nectar production of columnar cacti varied with latitude. Although this variation is positively correlated with pollination by diurnal visitors, it is influenced by phylogeny. The degree of specificity in the pollination systems of columnar cacti is heavily influenced by ecological factors and has a predictable geographic pattern. PMID- 21622311 TI - Systematics of the Carex aquatilis and C. lenticularis lineages: Geographically and ecologically divergent sister clades of Carex section Phacocystis (Cyperaceae). AB - Carex aquatilis is a highly diverse and geographically widespread member of one of the largest genera of flowering plants, Carex, and is ideally suited for the study of the role of hybridization and niche partitioning in ecological speciation. Phylogenetic analyses of nuclear ITS and ETS 1f and chloroplast psbA trnH DNA sequences support the monophyly of a broadly defined Carex aquatilis Carex lenticularis lineage, which includes C. aquatilis and C. lenticularis and their allies within section Phacocystis. However, neither taxon is monophyletic as currently circumscribed. The C. aquatilis lineage includes C. aquatilis and four morphologically and molecularly distinct salt-tolerant maritime taxa with which C. aquatilis s.s. is reported to form stabilized homoploid hybrids. The C. lenticularis lineage includes a paraphyletic C. lenticularis and seven allied species from both the New and Old World. The data provided here allow recognition of four species within the North American endemic C. lenticularis and suggest a neotropical origin for the C. lenticularis lineage with subsequent radiation and divergence through northwestern North America to Asia and via northeastern North America to Europe and southern South America. Evolutionary rate analyses indicate an origin for the C. aquatilis-C. lenticularis group around 1.89 million years ago during the early Pleistocene. PMID- 21622312 TI - The evolutionary origin of a second radiation of annual Castilleja (Orobanchaceae) species in South America: The role of long distance dispersal and allopolyploidy. AB - Considerable attention has been directed toward understanding the wide gaps in range that are common among many groups of closely related organisms. By placing their biology and geography in a phylogenetic context, we may gain a broader knowledge of the series of historical events that have led to present species distributions. In addition to the North American annuals, a second radiation of annual Castilleja species is in Andean Peru and central Chile. Phylogenetic analyses of chloroplast and nuclear DNA regions revealed a complex history for the origin and diversification of annual Castilleja species in South America. In addition to at least three independent long-distance dispersal events from North America, allopolyploidy has played a significant role in this disjunct radiation. Only C. attenuata occurs in both California and South America, and these results support its recent arrival to central Chile. Two Peruvian species are inferred to be allopolyploids; hybridization between annual lineages derived from independent long-distance dispersal events from North America gave rise to C. profunda, and hybridization between South American annual and perennial species gave rise to C. cerroana. The relative importance these events are discussed with reference to the observed morphological, ecological, and distributional patterns. PMID- 21622313 TI - Morphology and development of the gynoecium in Centrolepidaceae: The most remarkable range of variation in Poales. AB - This paper explores the relative impacts of reduction and polymerization on the evolution of reproductive structures in the small but morphologically diverse family Centrolepidaceae. Centrolepidaceae are closely related to Restionaceae and belong to the large order Poales, which also includes the grasses. In the largest genus of Centrolepidaceae, Centrolepis, the reproductive structures are viewed either as highly unusual aggregations of reduced flowers (the pseudanthial interpretation) or as unique flowers evolved through extreme reduction in the androecium, usually accompanied by a drastic increase in carpel number and elaboration of the entire gynoecium. Comparative data are here presented on gynoecia of all three genera of Centrolepidaceae; these data strongly support the latter (euanthial) interpretation. The combined phenomenon of carpel multiplication and decrease in stamen number is unexpected in a predominantly wind-pollinated lineage. Applying a pseudanthial interpretation would create a considerable morphological gap with reproductive structures of other Poales, whereas accepting a euanthial concept allows an almost continuous morphological series with related taxa. PMID- 21622314 TI - Developmental patterns in anatomy are shared among separate evolutionary origins of stem succulent and storage root-bearing growth habits in Adenia (Passifloraceae). AB - The architecture of flowering plants is astonishingly diverse. To understand evolutionary patterns and processes that account for this diversity, I investigated developmental anatomy of storage roots and stems of 58 species in the genus Adenia (Passifloraceae) using an explicit phylogenetic context. Because expanded storage roots and stem succulence evolved multiple times in Adenia, patterns of transition between succulent and nonsucculent forms were analyzed using a comparative test that accommodates phylogenetic uncertainty. I tested the innervation hypothesis, wherein I expected the evolution of vascular strands to be correlated with evolutionary increases in water storage tissue if evolution of vascular strands facilitates transport through water and starch storage structures. Not only is evolution of vascular strands in stems statistically coupled with evolutionary increases in parenchyma storage tissue, most lineages that evolved expanded storage roots also evolved vascular strands in these roots in parallel to succulent stems. I proposed that vascular strands and closely associated storage parenchyma found in both roots and shoots of Adenia comprise a homologous unit. A switch-like evolutionary mechanism that alters the spatial expression of this unit between roots and shoots can account, in large part, for transitions between markedly different habits such as storage-rooted herbs and succulent-stemmed shrubs. PMID- 21622315 TI - Reproductive short-shoots of Ginkgo biloba: A quantitative analysis of the disposition of axillary structures. AB - Ginkgo biloba, the only living representative in an otherwise extinct clade, is of pivotal importance to understanding seed plant phylogeny. Although G. biloba and its fossil relatives have been studied for over two centuries, there are both gaps and contradictions in the information available. We present data documenting the distributions of strobili and consider what an understanding of the disposition of strobili along short-shoots in Ginkgo adds to knowledge of the evolution of reproductive structures in seed plants in general. The megasporangiate strobili are found at and around the boundary between bracts and foliage leaves, while the expanse of microsporangiate strobili centers on the fifth bract back from that boundary. Quantitative analysis of the locations of the strobili along the short-shoot finds that increases in numbers of strobili are the result of recruitment of adjacent axils into morphogenetic activity. Gaps in the series of strobili are exceedingly rare. Further, while increased numbers of megasporangiate strobili arise from the symmetrical addition of axils into the fertile zone, increased numbers of microsporangiate strobili arise from a distinctly asymmetrical, basipetally biased, addition of axillary positions. This accurate morphological framework should orient molecular genetic studies that probe gymnosperm development itself or that consider gymnosperms as the proximate sources of gene expression redeployed in the origin of the angiosperm flower. PMID- 21622316 TI - Sex-specific variation in the interaction between Distichlis spicata (Poaceae) and mycorrhizal fungi. AB - Associations between mycorrhizal fungi and plants can influence intraspecific competition and shape plant population structure. While variation in plant genotypes is known to affect mycorrhizal colonization in crop systems, little is known about how genotypes affect colonization in natural plant populations or how plant sex might influence colonization with mycorrhizal fungi in plant species with dimorphic sexual systems. In this study, we analyzed mycorrhizal colonization in males and females of the wetland dioecious grass Distichlis spicata, which has spatially segregated sexes. Our results suggest that D. spicata males and females interact with mycorrhizal fungi differently. We discuss the implications for the role of this sex-specific symbiotic interaction in the maintenance of the within-population sex ratio bias of D. spicata. PMID- 21622317 TI - Fragment quality and matrix affect epiphytic performance in a Mediterranean forest landscape. AB - Destruction and fragmentation of habitats represent one of the most important threats for biodiversity. Here, we examined the effects of fragmentation in Mediterranean forests on the epiphytic lichen Lobaria pulmonaria (Lobariaceae). We tested the hypothesis that not only the level of fragmentation affects L. pulmonaria populations, but also the quality of the habitat and the nature of the surrounding matrix affect them. The presence and abundance of the lichen was recorded on 2039 trees in a total of 31 stands. We recorded habitat quality and landscape variables at three hierarchical levels: tree, plot, and patch. We found that L. pulmonaria tends to occur in trees with larger diameters in two types of surveyed forests. In Quercus pyrenaica patches, the mean diameter of colonized trees was smaller, suggesting the importance of bark roughness. Factors affecting the presence and cover of the lichen in each type of forest were different. There was a strong positive influence of distance from a river in beech forests, whereas proximity to forest edge positively affected in oak forests. The influence of the surrounding matrix was also an important factor explaining the epiphytic lichen abundance. PMID- 21622318 TI - Physiological integration in an introduced, invasive plant increases its spread into experimental communities and modifies their structure. AB - What determines the invasiveness of introduced plants is still poorly known. Many of the most invasive plant species are clonal, and physiological integration between connected individuals (ramets) of clonal plants may contribute to their ability to spread into communities and reduce performance of existing species. This contribution of integration to the invasiveness of clonal plants may be greater in denser communities. A greenhouse study was conducted to test these two hypotheses. High- and low-density communities were created by sowing seeds of eight grassland species. Each community was planted with three ramets of the stoloniferous, introduced plant Alternanthera philoxeroides that were disconnected from or left connected to ramets growing on bare soil. Connection increased the spread of Alternanthera within a community, but did not reduce community biomass. Alternanthera grew less in high-density communities, but connection did not improve its growth more than in low-density communities. Low density communities had higher evenness when Alternanthera was connected than when it was disconnected because shoot mass was lower in the more abundant species in the community and higher in the less abundant ones. These results partly supported the first hypothesis, but not the second. The effect of integration on community structure could be due to higher resource import by the ramets of Alternanthera closer to the dominant species. Integration therefore can increase the initial spread of new clonal plant species into communities and modify the effects of this spread on community structure. PMID- 21622319 TI - Kin recognition: Competition and cooperation in Impatiens (Balsaminaceae). AB - The ability to recognize kin is an important element in social behavior and can lead to the evolution of altruism. Recently, it has been shown that plants are capable of kin recognition through root interactions. Here we tested for kin recognition in a North American species of Impatiens that has a high opportunity of growing with kin and responds strongly to aboveground competition. We measured how the plants responded to the aboveground light quality cues of competition and to the presence of root neighbors and determined whether the responses depended on whether the neighbors were siblings or strangers. The study families were identified by DNA sequencing as members of the same species, provisionally identified as Impatiens pallida (hereafter I. cf. pallida). We found that I. cf. pallida plants were capable of kin recognition, but only in the presence of another plant's roots. Several traits responded to relatedness in shared pots, including increased leaf to root allocation with strangers and increased stem elongation and branchiness in response to kin, potentially indicating both increased competition toward strangers and reduced interference (cooperation) toward kin. Impatiens cf. pallida responded to both competition cues simultaneously, with the responses to the aboveground competition cue dependent on the presence of the belowground competition cue. PMID- 21622320 TI - Mycorrhizal diversity in Apostasia (Orchidaceae) indicates the origin and evolution of orchid mycorrhiza. AB - We demonstrated that "orchid mycorrhiza," a specialized mycorrhizal type, appeared in the common ancestor of the largest plant family Orchidaceae and that the fungal partner shifted from Glomeromycota to a particular clade of Basidiomycota in association with this character evolution. Several unique mycorrhizal characteristics may have contributed to the diversification of the family. However, the origin of orchid mycorrhiza and the diversity of mycobionts across orchid lineages still remain obscure. In this study, we investigated the mycorrhizae of five Apostasia taxa, members of the earliest-diverging clade of Orchidaceae. The results of molecular identification using nrDNA ITS and LSU regions showed that Apostasia mycorrhizal fungi belong to families Botryobasidiaceae and Ceratobasidiaceae, which fall within the order Cantharellales of Basidiomycota. Most major clades in Orchidaceae also form mycorrhizae with members of Cantharellales, while the sister group and other closely related groups to Orchidaceae (i.e., Asparagales except for orchids and the "commelinid" families) ubiquitously form symbioses with Glomeromycota to form arbuscular mycorrhizae. This pattern of symbiosis indicates that a major shift in fungal partner occurred in the common ancestor of the Orchidaceae. PMID- 21622321 TI - Do extreme environments provide a refuge from pathogens? A phylogenetic test using serpentine flax. AB - Abiotically extreme environments are often associated with physiologically stressful conditions, small, low-density populations, and depauperate flora and fauna relative to more benign settings. A possible consequence of this may be that organisms that occupy these stressful habitats receive fitness benefits associated with reductions in the frequency and/or intensity of antagonistic species interactions. I investigated a particular form of this effect, formalized as the "pathogen refuge hypothesis," through a study of 13 species of wild flax that grow on stressful serpentine soils and are often infected by a pathogenic fungal rust. The host species vary in the degree of their serpentine association: some specialize on extreme serpentine soils, while others are generalists that occur on soils with a wide range of serpentine influence. Phylogenetically explicit analyses of soil chemistry and field-measured disease levels indicated that rust disease was significantly less frequent and severe in flax populations growing in more stressful, low-calcium serpentine soils. These findings may help to explain the persistence of extremophile species in habitats where stressful physical conditions often impose strong autecological fitness costs on associated organisms. Ancestral state reconstruction of serpentine soil tolerance (approximated using soil calcium concentrations) suggested that the ability to tolerate extreme serpentine soils may have evolved multiple times within the focal genus. PMID- 21622322 TI - Genetic diversity of the endangered and narrow endemic Piperia yadonii (Orchidaceae) assessed with ISSR polymorphisms. AB - Highly endangered plants that are also narrow endemics are generally found to be genetically depauperate and thus are exceedingly susceptible to ecological and anthropological threats that can lead to their extinction. Piperia yadonii is restricted to a single California county within a biodiversity hotspot. We used nine primers to generate intersimple sequence repeat (ISSR) data to assess its genetic diversity and structure. Within each population, 99% of the loci were polymorphic, expected heterozygosity was low, and a majority of the loci were shared with few other populations. Forty percent of the total variation could be attributed to population differentiation while the rest (60%) resides within populations, and the genetic distances between populations were independent of the corresponding geographical distances. High divergence among populations is likely due to fragmentation and limited gene flow. Each population contains several private loci, and ideally, each should be protected to preserve the overall diversity of the species. Because P. yadonii currently retains a modest amount of genetic variation among individuals within populations, preserving and expanding the habitat at each site to allow natural expansion of populations would be additional strategies for its conservation before populations become too small to persist naturally. PMID- 21622323 TI - Papuacedrus (Cupressaceae) in Eocene Patagonia: A new fossil link to Australasian rainforests. AB - The 51.9 Ma Laguna del Hunco (LH) and 47.5 Ma Rio Pichileufu (RP) floras from Patagonia, Argentina are unusually rich, angiosperm-dominated assemblages with living relatives in the low-latitude West Pacific, neotropics, and temperate southern latitudes. The diverse gymnosperms in these floras are important for Gondwanan biogeographic history and paleoclimatic interpretations. "Libocedrus" prechilensis Berry 1938 (Cupressaceae), previously known only from the holotype (RP), a vegetative branch, is revised here based on new material from both localities, including a seed cone attached to a shoot with cuticle (LH). Characters of these fossils are diagnostic of monotypic Papuacedrus (highlands of New Guinea and Moluccas). Living P. papuana is most abundant in cloud forests receiving up to 4 m rainfall annually, whereas Austrocedrus (Libocedrus) chilensis, the basis of comparison when the fossil species was named, inhabits dry, cold steppe margins to mediterranean climates in southern South America. We establish Papuacedrus prechilensis comb. nov., which simultaneously invalidates a southern South American connection for the fossil floras and reveals a link to West Pacific montane rainforests. Combined evidence indicates a biome similar to extant subtropical, or tropical montane, rainforests that persisted for at least 4.4 Myr, linking elevated floral richness to abundant rainfall. PMID- 21622324 TI - Tertiary leaf fossils of Mangifera (Anacardiaceae) from Li Basin, Thailand as examples of the utility of leaf marginal venation characters. AB - The mango (Mangifera) is the most important commercial fruit in Asia and is popular worldwide. Because of uncertainty in its origin and biogeography, the study of Mangifera could be of value in elucidating its current genetic diversity and geographical distribution. We report here two new species of compressed leaves of Mangifera (Anacardiaceae) from upper Oligocene-lower Miocene lacustrine sediments in the Li Basin, northern Thailand. Leaf form and venation characters of fossil and extant leaves are presented with detailed marginal venation patterns that were found to be important in assigning the fossil leaves to Mangifera paleoindica and M. paleocaloneura. These two fossil species of Mangifera and the present occurrence of basal species of Mangifera in Thailand suggest that northern Thailand could be an area with a long history of evolution and diversification of Mangifera, particularly M. indica. PMID- 21622325 TI - The role of resources and architecture in modeling floral variability for the monoecious amphicarpic Emex spinosa (Polygonaceae). AB - Determining the sources of floral variation is crucial to the understanding of floral evolution. Architectural effects and phenotypic plasticity in development can play an important role in intraplant floral variation, giving rise to gender dimorphism or sexual specialization. Amphicarpic plants have another source of floral variation that could also be influenced by positional effects. We studied the effects of resource availability and architecture in intraplant floral variability in two ecotypes of the amphicarpic Emex spinosa. Male flowers were smaller than females, irrespective of position or resource availability. Emex spinosa shows gender dimorphism not influenced by positional effects. Flower size differences among positions were mainly due to architecture, because the effects of resources on flower size were minimal. Architectural effects caused a decrease in female flower size from ground to apical positions but an increase in most male traits that resulted in sexual specialization at the end of flowering. In general, the ecotypes were similarly affected by resources or architecture. Differences between subterranean and aerial female flowers seem also to be a consequence of architecture. Our results contribute to the evidence that resource limitation is an overestimated effect and that architectural effects must be considered in studies of floral or fruit variation. PMID- 21622326 TI - The maintenance of mixed mating by cleistogamy in the perennial violet Viola septemloba (Violaceae). AB - The production of both potentially outcrossed (chasmogamous) and obligately self fertilized (cleistogamous) flowers presents a clear exception to the prediction that the only evolutionarily stable mating systems are complete selfing and complete outcrossing. Although cleistogamy has evolved repeatedly, the reason for its stability is not known for any species. We tested the hypothesis that the production of cleistogamous and chasmogamous flowers by a perennial violet constitutes adaptive phenotypic plasticity. We manipulated the season of flowering for each flower type and determined fruit set and the germination percentage of seeds produced by cleistogamous and chasmogamous flowers to test the hypothesis that adaptive plastic response to seasonal environmental variation makes mixed mating stable. Cleistogamous flowers had greater fruit set in all seasons and produced seeds with germination percentages as great as or greater than those from chasmogamous flowers. The consistent advantage of cleistogamous flowers is clearly not consistent with a role of adaptive plastic response to seasonal variation. The biomass cost of seed production by chasmogamous flowers was nearly three times that for cleistogamous flowers. Explaining why chasmogamous flower have not been eliminated by natural selection requires that this difference be balanced by an advantage to chasmogamous flowers that has not yet been identified. PMID- 21622327 TI - Pollinators, "mustard oil" volatiles, and fruit production in flowers of the dioecious tree Drypetes natalensis (Putranjivaceae). AB - The Putranjivaceae is an enigmatic family, notable for being the only lineage outside the Capparales to possess the glucosinolate biochemical pathway, which forms the basis of an induced chemical defense system against herbivores (the "mustard oil bomb"). We investigated the pollination biology and floral scent chemistry of Drypetes natalensis (Putranjivaceae), a dioecious subcanopy tree with flowers borne on the stem (cauliflory). Flowering male trees were more abundant than female ones and produced about 10-fold more flowers. Flowers of both sexes produce copious amounts of nectar on disc-like nectaries accessible to short-tongued insects. The main flower visitors observed were cetoniid beetles, bees, and vespid wasps. Pollen load analysis indicated that these insects exhibit a high degree of fidelity to D. natalensis flowers. Insects effectively transfer pollen from male to female plants resulting in about 31% of female flowers developing fruits with viable seeds. Cetoniid beetles showed significant orientation toward the scent of D. natalensis flowers in a Y-maze olfactometer. The scents of male and female flowers are similar in chemical composition and dominated by fatty acid derivatives and isothiocyanates from the glucosinolate pathway. The apparent constitutive emission of isothiocyanates raises interesting new questions about their functional role in flowers. PMID- 21622328 TI - Evolutionary relationships, interisland biogeography, and molecular evolution in the Hawaiian violets (Viola: Violaceae). AB - The endemic Hawaiian flora offers remarkable opportunities to study the patterns of plant morphological and molecular evolution. The Hawaiian violets are a monophyletic lineage of nine taxa distributed across six main islands of the Hawaiian archipelago. To describe the evolutionary relationships, biogeography, and molecular evolution rates of the Hawaiian violets, we conducted a phylogenetic study using nuclear rDNA internal transcribed spacer sequences from specimens of each species. Parsimony, maximum likelihood (ML), and Bayesian inference reconstructions of island colonization and radiation strongly suggest that the Hawaiian violets first colonized the Maui Nui Complex, quickly radiated to Kaua'i and O'ahu, and recently dispersed to Hawai'i. The lineage consists of "wet" and "dry" clades restricted to distinct precipitation regimes. The ML and Bayesian inference reconstructions of shifts in habitat, habit, and leaf shape indicate that ecologically analogous taxa have undergone parallel evolution in leaf morphology and habit. This parallel evolution correlates with shifts to specialized habitats. Relative rate tests showed that woody and herbaceous sister species possess equal molecular evolution rates. The incongruity of molecular evolution rates in taxa on younger islands suggests that these rates may not be determined by growth form (or lifespan) alone, but may be influenced by complex dispersal events. PMID- 21622329 TI - Significance of ecological vicariance and long-distance dispersal in the diversification of Carex sect. Spirostachyae (Cyperaceae). AB - Plant disjunctions have provided one of the most intriguing distribution patterns historically addressed by biogeographers. Carex sect. Spirostachyae (Cyperaceae) displays an interesting pattern of disjunction to evaluate these scenarios, with species occurring in the main continental landmasses and in oceanic islands of the two hemispheres. Internal transcribed spacer and 5'-trnK intron plastid gene sequences were analyzed to determine (1) the times of diversification using penalized likelihood, and (2) reconstructions of the regions using maximum likelihood and Bayesian approaches of origin of sect. Spirostachyae and internal main lineages. The times for the diversification of sect. Spirostachyae are dated to between the end of the Eocene and the Oligocene, whereas the two main lineages are dated to between the end of the Oligocene and the beginning of Miocene. The phylogenetic analyses reveal a Mediterranean-Eurasian center of differentiation for sect. Spirostachyae and subsection Spirostachyae, whereas no clear, single ancestral area could be inferred for subsection Elatae. Both long-distance dispersal and ecological vicariance appear to have been involved in the evolutionary history of the disjunct distribution of the main lineages of sect. Spirostachyae. These organisms appear to have a special ability to colonize remote areas (through transoceanic and interhemispherical colonizations), but special long-distance dispersal mechanisms are not evident. PMID- 21622330 TI - The "sensational" power of movement in plants: A Darwinian system for studying the evolution of behavior. AB - Darwin's research on botany and plant physiology was a landmark attempt to integrate plant movements into a biological perspective of behavior. Since antiquity, people have sought to explain plant movements via mechanical or physiological forces, and yet they also constructed analogies between plant and animal behavior. During the Renaissance and Enlightenment, thinkers began to see that physiochemical explanations of plant movements could equally apply to animal behavior and even human thought. Darwin saw his research on plant movements as a strategic front against those who argued that his theory of evolution could not account for the acquisition of new behavioral traits. He believed that his research explained how the different forms of plant movement evolved as modified habits of circumnutation, and he presented evidence that plants might have a brain-like organ, which could have acquired various types of plant sensitivity during evolution. Upon publication of The Power of Movement in Plants, his ideas were overwhelmingly rejected by plant physiologists. Subsequently, plant biologists came to view the work as an important contribution to plant physiology and biology, but its intended contribution to the field of evolution and behavior has been largely overlooked. PMID- 21622331 TI - "The orchids have been a splendid sport"--an alternative look at Charles Darwin's contribution to orchid biology. AB - Charles Darwin's work with orchids and his thoughts about them are of great interest and not a little pride for those who are interested in these plants, but they are generally less well known than some of his other studies and ideas. Much has been published on what led to his other books and views. However, there is a paucity of information in the general literature on how Darwin's orchid book came about. This review will describe how The Various Contrivances by Which Orchids Are Fertilised by Insects came into being and will discuss the taxonomy of the orchids he studied. It also will concentrate on some of the less well-known aspects of Darwin's work and observations on orchids-namely, rostellum, seeds and their germination, pollination effects, and resupination-and their influence on subsequent investigators, plant physiology, and orchid science. PMID- 21622332 TI - Bark anatomy in Croton draco var. draco (Euphorbiaceae). AB - Many arboreal forms of the genus Croton (ca. 800 spp.), amply distributed in the Americas, have latex-producing cells in their bark, which is widely used in traditional medicine to treat skin infections and some forms of cancer. Studies validate its ethnomedicinal use-more than 20 pharmaceutically important secondary metabolites have been reported for its latex and bark-but anatomical and ecological studies are scarce. Given this species' ample distribution, laticifer abundance could be affected by the environment. We tested this for genetically similar trees growing in two types of vegetation in Veracruz, Mexico at sites commonly visited by traditional doctors. We describe the bark anatomy of C. draco, focusing on the laticifers, histochemically characterize the bark and the latex extracted from it, and document differences in laticifer abundance in the two environments. We have also identified another cell type (what we call type B) in the secretory latex system and describe it histochemically and microscopically. The location of bark cells that contain essential oils is reported here for the first time. Given the genetic similarity of the trees at both sites, the between-site variation in the number of laticifers in stem and branch bark appears to be an effect of the environment. PMID- 21622333 TI - Vessel grouping patterns in subfamilies Apocynoideae and Periplocoideae confirm phylogenetic value of wood structure within Apocynaceae. AB - This study contributes to our understanding of the phylogenetic significance and major evolutionary trends in the wood of the dogbane family (Apocynaceae), one of the largest and economically most important angiosperm families. Based on LM and SEM observations of 56 Apocynoideae species-representing all currently recognized tribes-and eight Periplocoideae, we found striking differences in vessel grouping patterns (radial multiples vs. large clusters) between the mainly nonclimbing apocynoid tribes (Wrightieae, Malouetieae, Nerieae) and the climbing lineages (remaining Apocynoideae and Periplocoideae). The presence of large vessel clusters in combination with fibers in the ground tissue characterizing the climbing Apocynoideae and Periplocoideae clearly contrasts with the climbing anatomy of the rauvolfioids (solitary vessels plus tracheids in ground tissue), supporting the view that (1) the climbing habit has evolved more than once in Apocynaceae, (2) the three nonclimbing apocynoid tribes are basal compared to the climbing apocynoids, and (3) Periplocoideae belong to the crown clade. The wood anatomy within the nonclimbing and climbing lineages is rather homogeneous, although a combination of specific characters (e.g. presence of septate fibers, axial parenchyma distribution, abundance of uniseriate compared to multiseriate rays, and presence and location of prismatic crystals) may be used to identify several tribes. PMID- 21622334 TI - Development of a complex floral trait: The pollinator-attracting petal spots of the beetle daisy, Gorteria diffusa (Asteraceae). AB - Angiosperms possess a variety of complex floral traits that attract animal pollinators. Dark petal spots have evolved independently many times across the angiosperm phylogeny and have been shown to attract insect pollinators from several lineages. Here we present new data on the ontogeny and morphological complexity of the elaborate insect-mimicking petal spots of the South African daisy species, Gorteria diffusa (Asteraceae), commonly known as the beetle daisy, although it is fly-pollinated. Using light and scanning electron microscopy and histology, we identified three distinct specialized cell types of the petal epidermis that compose the petal spot. Sophisticated patterning of pigments, cuticular elaborations, and multicellular papillate trichomes make the G. diffusa petal spot a uniquely complex three-dimensional floral ornament. Examination of young inflorescence meristems revealed that G. diffusa ray florets develop (and probably also initiate) basipetally, in the opposite direction to the disc florets-a developmental phenomenon that has been found in some other daisies, but which contradicts conventional theories of daisy inflorescence architecture. Using these ontogenetic and morphological data, we have identified the mechanism by which G. diffusa patterns its insect-mimicking petal spots, and we propose a testable model for the genetic regulation of petal spot identity. PMID- 21622335 TI - Cryptic bracts facilitate herbivore avoidance in the mycoheterotrophic plant Monotropsis odorata (Ericaceae). AB - Plant coloration, shown to play a dynamic role in animal attraction, has been proposed as a means of defense, although these reports lack experimental evidence. This study empirically assesses defensive coloration in the mycoheterotrophic plant, Monotropsis odorata, which produces stems and flowers covered by dried vegetative bracts. Field studies were conducted using an experimental group of plants with bracts removed and a control group to evaluate the frequency of and fitness impacts associated with herbivory. Additionally, we quantitatively assessed the reflectance spectra of bracts, stems, and flowers of M. odorata relative to an ambient leaf litter substrate. Across the 2-yr study, the experimental group experienced a 20-27% higher mean herbivory rate and 7-20% lower mean fruit production relative to the control group. Bracts were shown to strongly resemble ambient leaf litter in spectral analyses, with stems and flowers having more conspicuous coloration. Results show that the presence of dried bracts effectively camouflages conspicuous stem and floral tissues, significantly reducing the frequency of floral and stem herbivory, and thereby increasing fruit set, a component of plant fitness. This study supports the principal hypothesis that coloration can play a fundamental role in plant defense. PMID- 21622336 TI - Community development following gamma radiation at a pine-oak forest, Brookhaven National Laboratory, Long Island, New York1. AB - We investigated a unique source of forest disturbance: gamma radiation. While the temporal patterns of ecological succession are well understood for the forests of eastern North America, this is not the case for massively irradiated forests. Our objective was to compare vascular plant community change after irradiation at the five vegetation zones described in 1962 by Woodwell at Brookhaven National Laboratory, Long Island, New York. No follow-up studies have been done since the gamma radiation experiments were terminated in 1978. Ecological successional theory (e.g., Bormann and Likens, 1994, Likens and Bormann, 1995) does not explain long-term forest recovery after radiation damage. Our null hypothesis was that 47 yr after initial gamma ray exposure, the sites would have recovered such that floristic composition would be the same as the pine-oak forest control. This hypothesis was rejected statistically. In 2007/2008, the five concentric zones of vegetation centered about the gamma source retained their floristic heterogeneity as measured by Jaccard coefficients. PMID- 21622337 TI - Coordination of foliar and wood anatomical traits contributes to tropical tree distributions and productivity along the Malay-Thai Peninsula. AB - Drought is a critical factor in plant species distributions. Much research points to its relevance even in moist tropical regions. Recent studies have begun to elucidate mechanisms underlying the distributions of tropical tree species with respect to drought; however, how such desiccation tolerance mechanisms correspond with the coordination of hydraulic and photosynthetic traits in determining species distributions with respect to rainfall seasonality deserves attention. In the present study, we used a common garden approach to quantify inherent differences in wood anatomical and foliar physiological traits in 21 tropical tree species with either widespread (occupying both seasonal and aseasonal climates) or southern (restricted to aseasonal forests) distributions with respect to rainfall seasonality. Use of congeneric species pairs and phylogenetically independent contrast analyses allowed examination of this question in a phylogenetic framework. Widespread species opted for wood traits that provide biomechanical support and prevent xylem cavitation and showed associated reductions in canopy productivity and consequently growth rates compared with southern species. These data support the hypothesis that species having broader distributions with respect to climatic variability will be characterized by traits conducive to abiotic stress tolerance. This study highlights the importance of the well-established performance vs. stress tolerance trade-off as a contributor to species distributions at larger scales. PMID- 21622338 TI - Genetic diversity, structure, and demographic change in tanoak, Lithocarpus densiflorus (Fagaceae), the most susceptible species to sudden oak death in California. AB - Knowledge of population genetic structure of tanoak (Lithocarpus densiflorus) is of interest to pathologists seeking natural variation in resistance to sudden oak death disease, to resource managers who need indications of conservation priorities in this species now threatened by the introduced pathogen (Phytophthora ramorum), and to biologists with interests in demographic processes that have shaped plant populations. We investigated population genetic structure using nuclear and chloroplast DNA (cpDNA) and inferred the effects of past population demographic processes and contemporary gene flow. Our cpDNA results revealed a strong pattern of differentiation of four regional groups (coastal California, southern Oregon, Klamath mountains, and Sierra Nevada). The chloroplast haplotype phylogeny suggests relatively deep divergence of Sierra Nevada and Klamath populations from those of coastal California and southern Oregon. A widespread coastal California haplotype may have resulted from multiple refugial sites during the Last Glacial Maximum or from rapid recolonization from few refugia. Analysis of nuclear microsatellites suggests two major groups: (1) central coastal California and (2) Sierra Nevada/Klamath/southern Oregon and an area of admixture in north coastal California. The low level of nuclear differentiation is likely to be due to pollen gene flow among populations during postglacial range expansion. PMID- 21622339 TI - Two independent C4 origins in Aristidoideae (Poaceae) revealed by the recruitment of distinct phosphoenolpyruvate carboxylase genes. AB - Determining the number of evolutions of an adaptive novelty is primordial to understand its evolutionary significance. C(4) photosynthesis, an adaptation to low CO(2) atmospheric concentration and high temperature, evolved multiple times, but the number of convergent evolutions is still debated. In Poaceae phylogeny, numerous C(4) groups are separated by C(3) taxa, but whether these correspond to independent C(4) origins or a few C(4) evolutions followed by reversals is controversial. The Aristidoideae subfamily is formed by two C(4) genera, Aristida and Stipagrostis, separated by the C(3) genus Sartidia. In the current study, we investigated the evolutionary history of genes encoding phosphoenolpyruvate carboxylases (PEPC) to shed light on the photosynthetic transitions that occurred in Aristidoideae. We identified six distinct PEPC gene lineages that appeared through several rounds of gene duplications before or early during grass diversification. The gene lineage encoding the C(4) PEPC of Stipagrostis differs from those of the other C(4) grasses, including Aristida. These distinct origins of C(4) PEPC genes from these two Aristidoideae genera unequivocally indicate that they integrated the C(4) pathway independently. This highlights the importance of candidate-gene studies when inferring the evolutionary history of a character such as C(4) photosynthesis, one of the greatest evolutionary successes in plant history. PMID- 21622340 TI - New insights into the phylogenetic relationships, character evolution, and phytogeographic patterns of Calceolaria (Calceolariaceae). AB - Biogeographical patterns and diversification processes in Andean and Patagonian flora are not yet well understood. Calceolaria is a highly diversified genus of these areas, representing one of the most specialized plant-pollinator systems because flowers produce nonvolatile oils, a very unusual floral reward. Phylogenetic analyses with molecular (ITS and matK) and morphological characters from 103 Calceolaria species were conducted to examine relationships, to understand biogeographic patterns, and to detect evolutionary patterns of floral and ecological characters. Total evidence analysis retrieved three major clades, which strongly correspond to the three previously recognized subgenera, although only subgenus Rosula was retrieved as a monophyletic group. A single historical event explains the expansion from the southern to central Andes, while different parallel evolutionary lines show a northward expansion from the central to northern Andes across the Huancabamba Deflection, an important geographical barrier in northern Peru. Polyploidy, acquisition of elaiophores, and a nototribic pollination mechanism are key aspects of the evolutionary history of Calceolaria. Pollination interactions were more frequently established with Centris than with Chalepogenus oil-collecting bee species. The repeated loss of the oil gland and shifts to pollen as the only reward suggest an evolutionary tendency from highly to moderately specialized pollination systems. PMID- 21622341 TI - An Early Cretaceous fruit with affinities to Ceratophyllaceae. AB - A new genus and species with affinities to Ceratophyllaceae from the Lower Cretaceous of Kansas, USA is reported. The fruits of Donlesia dakotensis gen. et sp. nov. are smooth achenes with two winged lateral spines, two winged facial spines, one stylar spine, and a long peduncle with a distinctive groove. The facial spines are arranged perpendicular to the lateral spines. The seed contains two cotyledons and a central plumule. Dichotomous leaves associated with Donlesia dakotensis fruits are whorled, petiolate, and lack denticles. The fruit morphology of D. dakotensis differs from extant and fossil members of Ceratophyllaceae in its tetra-radial symmetry, the presence of two winged facial spines, a long peduncle, and the lack of other appendages on the fruit surface. It seems to be more closely related to section Ceratophyllum of extant Ceratophyllum. This relationship suggests that section Ceratophyllum of extant Ceratophyllum may represent the ancestral members of the ancient Ceratophyllum lineage. The presence of this species in the Dakota Formation further confirms that there were diverse aquatic plants living in freshwater lakes along the eastern margin of the Western Interior Seaway during the Early Cretaceous. PMID- 21622342 TI - Pollination biology of Harrisia portoricensis (Cactaceae), an endangered Caribbean species. AB - Traits associated with self-pollination are common features of island plant communities. In this work, we studied the pollination biology and the breeding system of Harrisia portoricensis, an island columnar cactus, to test for the presence of inbreeding and inbreeding depression. For H. portoricensis, which bears flowers with typical outcrossing morphology, the results from 322 h of direct observations and videotaping showed that visits to flowers by animals were uncommon. Controlled pollinations demonstrated that H. portoricensis has a partially self-compatible breeding system that it is not autogamous and thus requires an external mechanism for the movement of pollen to set fruit. We detected differences in seed size, seed mass, germination success, and multiplicative fitness estimates between self- and cross-pollination treatments. We found that progeny resulting from natural and self-pollination treatments showed signs of inbreeding depression compared with progeny resulting from cross pollination; however, the magnitude of the inbreeding depression was less than 50%. Our combined results suggest that for this species an endogamous breeding system should be favored by natural selection. PMID- 21622343 TI - Early inbreeding depression in the sexually polymorphic plant Dianthus sylvestris (Caryophyllaceae): Effects of selfing and biparental inbreeding among sex morphs. AB - Predominantly outcrossing plant species are expected to accumulate recessive deleterious mutations, which can be purged when in a homozygous state following selfing. Individuals may vary in their genetic load because of different selfing histories, which could lead to differences in inbreeding depression among families. Lineage-dependent inbreeding depression can appear in gynodioecious species if obligatory outcrossed females are more likely to produce female offspring and if partially selfing hermaphrodites are more likely to produce hermaphrodites. We investigated inbreeding depression at the zygote, seed, and germination stages in the gynomonoecious-gynodioecious Dianthus sylvestris, including pure-sexed plants and a mixed morph. We performed hand-pollinations on 56 plants, belonging to the three morphs, each receiving 2-3 cross treatments (out-, sib- and self-pollination) on multiple flowers. Effects of cross treatments varied among stages and influenced seed provisioning, with sibling competition mainly occurring within outcrossed fruits. We found significant inbreeding depression for seed mass and germination and cumulative early inbreeding depression varied greatly among families. Among sex morphs, we found that females and hermaphrodites differed in biparental inbreeding depression, whereas uniparental was similar for all. Significant inbreeding depression levels may play a role in female maintenance in this species, and individual variation in association with sex-lineages proclivity is discussed. PMID- 21622344 TI - Seed fertilization, development, and germination in Hydatellaceae (Nymphaeales): Implications for endosperm evolution in early angiosperms. AB - New data on endosperm development in the early-divergent angiosperm Trithuria (Hydatellaceae) indicate that double fertilization results in formation of cellularized micropylar and unicellular chalazal domains with contrasting ontogenetic trajectories, as in waterlilies. The micropylar domain ultimately forms the cellular endosperm in the dispersed seed. The chalazal domain forms a single-celled haustorium with a large nucleus; this haustorium ultimately degenerates to form a space in the dispersed seed, similar to the chalazal endosperm haustorium of waterlilies. The endosperm condition in Trithuria and waterlilies resembles the helobial condition that characterizes some monocots, but contrasts with Amborella and Illicium, in which most of the mature endosperm is formed from the chalazal domain. The precise location of the primary endosperm nucleus governs the relative sizes of the chalazal and micropylar domains, but not their subsequent developmental trajectories. The unusual tissue layer surrounding the bilobed cotyledonary sheath in seedlings of some species of Trithuria is a belt of persistent endosperm, comparable with that of some other early-divergent angiosperms with a well-developed perisperm, such as Saururaceae and Piperaceae. The endosperm of Trithuria is limited in size and storage capacity but relatively persistent. PMID- 21622345 TI - The filifolium1 mutation perturbs shoot architecture in Zea mays (Poaceae). AB - Plant architecture is elaborated through the activity of shoot apical meristems (SAMs), which produce repeating units known as phytomers, that are comprised of leaf, node, internode, and axillary bud. Insight into how SAMs function and how individual phytomer components are related to each other can been obtained through characterization of recessive mutants with perturbed shoot development. In this study, we characterized a new mutant to further understand mechanisms underlying shoot development in maize. The filifolium1-0 (ffm1-0) mutants develop narrow leaves on dwarfed shoots. Shoot growth often terminates at the seedling stage from depletion of the SAM, but if plants survive to maturity they are invariably bushy. KN1-like homeobox (KNOX) proteins are inappropriately regulated in mutant apices, adaxial identity is not specified in mutant leaves, and axillary meristems develop precociously. We propose that FFM1 acts to demarcate zones within the SAM so that appropriate fates can be conferred on cells within those zones by other factors. On the basis of the mutant phenotype, we also speculate about different relationships between phytomer components in maize and Arabidopsis. PMID- 21622346 TI - Limited flooding tolerance of juveniles restricts the distribution of adults in an understory shrub (Itea virginica; Iteaceae). AB - Juvenile plants often have tight microhabitat associations because of specific requirements for seed germination and subsequent establishment. Due to their larger size, adults may be more adept at coping with stress. However, few studies consider the role of ontogeny in structuring plant populations, even though phenotypic expression can change through life history. In cypress-tupelo swamps of the United States, understory species often grow on microsites above floodwaters. In a field survey of one such species, Itea virginica, we found that more than 98% of seedlings occurred on elevated microsites, which were relatively infrequent. However, this strict association relaxed through ontogeny, with nearly 8% of subadults and adults rooted directly on the forest floor. We hypothesized that flooding inhibits juvenile establishment on the forest floor. In greenhouse experiments, we investigated the effects of flooding and substrate on Itea performance. Seeds had similar germination rates on drained swamp soil and cypress knee wood. Seedling growth was high on unflooded soil, but declined precipitously when submerged. Finally, performance of seedlings, but not older plants, decreased with flood severity. Our results highlight the importance of assessing stress tolerance over multiple life history stages because limitations of juveniles can constrain the distribution patterns of future ontogenetic stages. PMID- 21622347 TI - Construction costs, payback times, and the leaf economics of carnivorous plants. AB - Understanding how different plant species and functional types "invest" carbon and nutrients is a major goal of plant ecologists. Two measures of such investments are "construction costs" (carbon needed to produce each gram of tissue) and associated "payback times" for photosynthesis to recover construction costs. These measurements integrate among traits used to assess leaf-trait scaling relationships. Carnivorous plants are model systems for examining mechanisms of leaf-trait coordination, but no studies have measured simultaneously construction costs of carnivorous traps and their photosynthetic rates to determine payback times of traps. We measured mass-based construction costs (CC(mass)) and photosynthesis (A(mass)) for traps, leaves, roots, and rhizomes of 15 carnivorous plant species grown under greenhouse conditions. There were highly significant differences among species in CC(mass) for each structure. Mean CC(mass) of carnivorous traps (1.14 +/- 0.24 g glucose/g dry mass) was significantly lower than CC(mass) of leaves of 267 noncarnivorous plant species (1.47 +/- 0.17), but all carnivorous plants examined had very low A(mass) and thus, long payback times (495-1551 h). Our results provide the first clear estimates of the marginal benefits of botanical carnivory and place carnivorous plants at the "slow and tough" end of the universal spectrum of leaf traits. PMID- 21622348 TI - Pollination context effects in the high-mountain dimorphic Armeria caespitosa (Plumbaginaceae): Neighborhood is something more than density. AB - Frequency-dependent processes are relevant for flowering plant reproduction, especially for species with disassortative mating. In an individual-based study, we tested not only the effects of local density on reproductive success at small spatial scales, but also those of neighborhood quality. To test the neighborhood effects on the reproduction of Armeria caespitosa, a dimorphic Mediterranean high mountain endemic, we introduce a novel pollination context (PC) index that considered the distance, floral display, and floral morph of neighbors at small scales (within 2 m from the focal plant), studying rock and pasture populations at both edges of the species altitudinal distribution. Reproductive success depended significantly on PC only at the low populations, suggesting that the PC effects are population-dependent and supporting the hypothesis that the neighborhood quality influences the reproductive success of A. caespitosa at least in stressful conditions. Moreover, fruit set was morph-dependent in the high-pasture population. The specific role of the spatial structure of compatible vs. incompatible morphs at small scales of the dimorphic self-incompatibility system in Armeria deserves further attention. Parameters other than plant density are useful for the study of small-scale density-dependent processes that affect pollination and other reproductive components, especially if they integrate neighborhood quality information at adequate spatial scales. PMID- 21622349 TI - Molecular evidence of reticulate evolution in the subgenus Plantago (Plantaginaceae). AB - Polyploidization is a frequent evolutionary event in plants that has a large influence on speciation and evolution of the genome. Molecular phylogenetic analyses of the taxonomically complex subgenus Plantago were conducted to elucidate intrasubgeneric phylogenetic relationships. A nuclear-encoding single copy gene, SUC1 (1.0-1.8 kb), was sequenced in 24 taxa representing all five sections of the subgenus Plantago and two taxa from subgenus Coronopus as the outgroup. Fifteen known polyploids and one putative polyploid were sampled to examine polyploid origins and occurrence of reticulate evolution by cloning and sequence analysis of SUC1. Phylogenetic relationships were estimated using maximum parsimony, neighbor-joining, and Bayesian analyses. For the first time, our analysis provides a highly resolved phylogenetic tree. Subgenus Plantago formed a well-supported monophyletic clade. In contrast, alleles from polyploid species were scattered across the whole SUC1 phylogenetic tree, and some independent allopolyploids originated from hybridization between distant lineages. One reason for this taxonomic complexity can be attributed to reticulate evolution within the subgenus Plantago. Our results also suggest the possibility of two independent long-distance dispersals between the northern and southern hemispheres. PMID- 21622350 TI - Deciphering the origins of apomictic polyploids in the Cheilanthes yavapensis complex (Pteridaceae). AB - Deciphering species relationships and hybrid origins in polyploid agamic species complexes is notoriously difficult. In this study of cheilanthoid ferns, we demonstrate increased resolving power for clarifying the origins of polyploid lineages by integrating evidence from a diverse selection of biosystematic methods. The prevalence of polyploidy, hybridization, and apomixis in ferns suggests that these processes play a significant role in their evolution and diversification. Using a combination of systematic approaches, we investigated the origins of apomictic polyploids belonging to the Cheilanthes yavapensis complex. Spore studies allowed us to assess ploidy levels; plastid and nuclear DNA sequencing revealed evolutionary relationships and confirmed the putative progenitors (both maternal and paternal) of taxa of hybrid origin; enzyme electrophoretic evidence provided information on genome dosage in allopolyploids. We find here that the widespread apomictic triploid, Cheilanthes lindheimeri, is an autopolyploid derived from a rare, previously undetected sexual diploid. The apomictic triploid Cheilanthes wootonii is shown to be an interspecific hybrid between C. fendleri and C. lindheimeri, whereas the apomictic tetraploid C. yavapensis is comprised of two cryptic and geographically distinct lineages. We show that earlier morphology-based hypotheses of species relationships, while not altogether incorrect, only partially explain the complicated evolutionary history of these ferns. PMID- 21622351 TI - The genomic organization of Ty3/gypsy-like retrotransposons in Helianthus (Asteraceae) homoploid hybrid species. AB - The origin of new diploid, or homoploid, hybrid species is associated with rapid genomic restructuring in the hybrid neospecies. This mode of speciation has been best characterized in wild sunflower species in the genus Helianthus, where three homoploid hybrid species (H. anomalus, H. deserticola, and H. paradoxus) have independently arisen via ancient hybridization events between the same two parental species (H. annuus and H. petiolaris). Most previous work examining genomic restructuring in these sunflower hybrid species has focused on chromosomal rearrangements. However, the origin of all three homoploid hybrid sunflower species also is associated with massive proliferation events of Ty3/gypsy-like retrotransposons in the hybrid species' genomes. We compared the genomic organization of these elements in the parent species and two of the homoploid hybrid species using fluorescence in situ hybridization (FISH). We found a significant expansion of Ty3/gypsy-like retrotransposons confined to the pericentromeric regions of two hybrid sunflower species, H. deserticola and H. paradoxus. In contrast, we detected no significant increase in the frequency or extent of dispersed retrotransposon populations in the hybrid species within the resolution limits of our assay. We discuss the potential role that transposable element proliferation and localization plays in the evolution of homoploid hybrid species. PMID- 21622352 TI - Mitotic instability in resynthesized and natural polyploids of the genus Arabidopsis (Brassicaceae). AB - Allopolyploids contain complete sets of chromosomes from two or more different progenitor species. Because allopolyploid hybridization can lead to speciation, allopolyploidy is an important mechanism in evolution. Meiotic instability in early-generation allopolyploids contributes to high lethality, but less is known about mitotic fidelity in allopolyploids. We compared mitotic stability in resynthesized Arabidopsis suecica-like neoallopolyploids with that in 13 natural lines of A. suecica (2n = 4x = 26). We used fluorescent in situ hybridization to distinguish the chromosomal contribution of each progenitor, A. thaliana (2n = 2x =10) and A. arenosa (2n = 4x = 32). Surprisingly, cells of the paternal parent A. arenosa had substantial aneuploidy, while cells of the maternal parent A. thaliana were more stable. Both natural and resynthesized allopolyploids had low to intermediate levels of aneuploidy. Our data suggest that polyploidy in Arabidopsis is correlated with aneuploidy, but varies in frequency by species. The chromosomal composition in aneuploid cells within individuals was variable, suggesting somatic mosaicisms of cell lineages, rather than the formation of distinct, stable cytotypes. Our results suggest that somatic aneuploidy can be tolerated in Arabidopsis polyploids, but there is no evidence that this type of aneuploidy leads to stable novel cytotypes. PMID- 21622353 TI - Tuberculate ectomycorrhizae of angiosperms: The interaction between Boletus rubropunctus (Boletaceae) and Quercus species (Fagaceae) in the United States and Mexico. AB - Tuberculate ectomycorrhizae (TECM) are unique structures in which aggregates of ectomycorrhizal roots are encased in a covering of fungal hyphae. The function of TECM is unknown, but they probably enhance the nitrogen nutrition and disease resistance of host plants. Trees in the Pinaceae form TECM with species of Rhizopogon and Suillus (Suillineae, Boletales). Similar tubercules are found with diverse angiosperms, but their mycobionts have not been phylogenetically characterized. We collected TECM in Mexico and the USA that were similar to TECM in previous reports. We describe these TECM and identify both the plant and fungal symbionts. Plant DNA confirms that TECM hosts are Quercus species. ITS sequences from tubercules and sclerotia (hyphal aggregations that serve as survival structures) matched sporocarps of Boletus rubropunctus. Phylogenetic analyses confirm that this fungus belongs to the suborder Boletineae (Boletales). This is the first published report of TECM formation in the Boletineae and of sclerotia formation by a Boletus species. Our data suggest that the TECM morphology is an adaptive feature that has evolved separately in two suborders of Boletales (Suillineae and Boletineae) and that TECM formation is controlled by the mycobiont because TECM are found on distantly related angiosperm and gymnosperm host plants. PMID- 21622354 TI - Paratingia wudensis sp. nov., a whole noeggerathialean plant preserved in an earliest Permian air fall tuff in Inner Mongolia, China. AB - Noeggerathiales are a little known group of Carboniferous and Permian plants of uncertain systematic position that have been variously considered to be ferns, sphenopsids, progymnosperms, or a separate group. These heterosporous plants carry adaxial sporangia on leaf-like or disk-shaped sporophylls that form cones. Leaves are pinnate with a rather stiff appearance, and pinnules can be attached in either two or four rows. In the present report, we present the top of a noeggerathialean plant with leaves and strobili attached, Paratingia wudensis Wang, Pfefferkorn et Bek sp. nov., from an earliest Permian volcanic ash fall tuff in Inner Mongolia. The excellent preservation allows the reconstruction of the whole plant, the complex three-dimensional leaves with anisophyllous pinnules, the heterosporous strobili, and the spores in situ. The homology of leaves and strobili can be elucidated and contributes to an understanding of the debated taxonomic position of Noeggerathiales. The "anisophyllous" leaves carry pinnules arranged in four rows. The strobili are bisporangiate and have disk shaped sporophylls, each with one ring of 10-14 adaxial sporangia around the strobilus axis. Megaspores have an equatorial bulge. This new species expands the known diversity of Noeggerathiales. It grew in a peat-forming forest, thus changing earlier interpretations of the growth of noeggerathialean plants with anisophyllous pinnules. PMID- 21622355 TI - Molecular, morphological, and ecological niche differentiation of sympatric sister oak species, Quercus virginiana and Q. geminata (Fagaceae). AB - The genus Quercus (the oaks) is notorious for interspecific hybrization, generating questions about the mechanisms that permit coexistence of closely related species. Two sister oak species, Quercus virginiana and Q. geminata, occur in sympatry in Florida and throughout the southeastern United States. In 11 sites from northern and southeastern regions of Florida, we used a leaf-based morphological index to identify individuals to species. Eleven nuclear microsatellite markers significantly differentiated between the species with a high correspondence between molecular and morphological typing of specimens. Nevertheless, Bayesian clustering analysis indicates interspecific gene flow, and six of 109 individuals had mixed ancestry. The identity of several individuals also was mismatched using molecular markers and morphological characters. In a common environment, the two species performed differently in terms of photosynthetic performance and growth, corresponding to their divergent ecological niches with respect to soil moisture and other edaphic properties. Our data support earlier hypotheses that divergence in flowering time causes assortative mating, allowing these ecologically distinct sister species to occur in sympatry. Limited gene flow that permits ecological differentiation helps to explain the overdispersion of oak species in local communities. PMID- 21622356 TI - Pollen clumping and wind dispersal in an invasive angiosperm. AB - Pollen dispersal is a fundamental aspect of plant reproductive biology that maintains connectivity between spatially separated populations. Pollen clumping, a characteristic feature of insect-pollinated plants, is generally assumed to be a detriment to wind pollination because clumps disperse shorter distances than do solitary pollen grains. Yet pollen clumps have been observed in dispersion studies of some widely distributed wind-pollinated species. We used Ambrosia artemisiifolia (common ragweed; Asteraceae), a successful invasive angiosperm, to investigate the effect of clumping on wind dispersal of pollen under natural conditions in a large field. Results of simultaneous measurements of clump size both in pollen shedding from male flowers and airborne pollen being dispersed in the atmosphere are combined with a transport model to show that rather than being detrimental, clumps may actually be advantageous for wind pollination. Initial clumps can pollinate the parent population, while smaller clumps that arise from breakup of larger clumps can cross-pollinate distant populations. PMID- 21622357 TI - An experimental demonstration of the cost of sex and a potential resource limitation on reproduction in the moss Pterygoneurum (Pottiaceae). AB - The cost of sexual reproduction is incurred when the current reproductive episode contributes to a a decline in future plant performance. To test the hypotheses that a trade-off exists between current sexual reproduction and subsequent clonal regeneration and that resources limit reproduction and regeneration, plants of the widespread moss Pterygoneurum ovatum were subjected to induced sporophytic abortion, upper leaf removal, and nutrient amendment treatments. Sexually reproducing plants were slower or less likely to produce regenerative structures (protonemata or shoots) and produced fewer regenerative tissue areas or structures. The ability and the timeline to reproduce sexually and regenerate clonally were unaffected by an inorganic nutrient amendment. However, when leaves subtending the sporophyte were removed, the sporophytes were less likely to mature, tended to take a longer time to mature, and were smaller compared to sporophytes from shoots with a full complement of upper leaves. Our findings indicate that plants investing in sexual reproduction suffer a cost of decreased clonal regeneration and indicate that sporophyte maturation is resource-limited, with upper leaves contributing to the nutrition of the sporophyte. This study represents only the second explicit experimental demonstration of a trade-off between sexual and asexual reproduction in bryophytes. PMID- 21622358 TI - Beetle pollination of the fruit-scented cones of the South African cycad Stangeria eriopus. AB - There has been considerable uncertainty about the importance of wind vs. insects in cycad pollination, but recent studies in several cycad genera have indicated that these are pollinated primarily, if not exclusively, by insects. Stangeria represents an isolated southern African cycad lineage previously thought to be wind-pollinated. Unlike in most other cycads, there is no evidence of cone thermogenesis in Stangeria. We found that the scent of both male and female Stangeria cones mimics that of fermented fruit, the main volatiles being esters of acetic acid, ketones, and aldehydes. We found a large variety of insect visitors on the cones, the most common ones being sap and rove beetles (Coleoptera: Nitidulidae, Staphylinidae) and fruit flies (Diptera: Drosophilidae). Of these, only sap beetles (Nitidulidae) were able to effect pollination under experimental conditions. Because sap beetles are also pollinators of Cycas and members of several ancient angiosperm families, their role in the pollination of Stangeria adds interesting details to the role this group of insects has played in the history of plant-pollinator interactions. PMID- 21622359 TI - A molecular phylogeny and classification of Bignoniaceae. AB - Bignoniaceae are woody, trees, shrubs, and lianas found in all tropical floras of the world with lesser representation in temperate regions. Phylogenetic analyses of chloroplast sequences (rbcL, ndhF, trnL-F) were undertaken to infer evolutionary relationships in Bignoniaceae and to revise its classification. Eight clades are recognized as tribes (Bignonieae, Catalpeae, Coleeae, Crescentieae, Jacarandeae, Oroxyleae, Tecomeae, Tourrettieae); additional inclusive clades are named informally. Jacarandeae and Catalpeae are resurrected; the former is sister to the rest of the family, and the latter occupies an unresolved position within the "core" Bignoniaceae. Tribe Eccremocarpeae is included in Tourrettieae. Past classifications recognized a large Tecomeae, but this tribe is paraphyletic with respect to all other tribes. Here Tecomeae are reduced to a clade of approximately 12 genera with a worldwide distribution in both temperate and tropical ecosystems. Two large clades, Bignonieae and Crescentiina, account for over 80% of the species in the family. Coleeae and Crescentieae are each included in larger clades, the Paleotropical alliance and Tabebuia alliance, respectively; each alliance includes a grade of taxa assigned to the traditional Tecomeae. Parsimony inference suggests that the family originated in the neotropics, with at least five dispersal events leading to the Old World representatives. PMID- 21622360 TI - Cuscuta jepsonii (Convolvulaceae): An invasive weed or an extinct endemic? AB - Despite their ecological significance, parasitic plants face more conservation challenges than do autotrophic plants. This is especially true for the groups that include weedy or invasive species such as Cuscuta. While approximately half of the Cuscuta (dodders) species may require conservation measures, the genus as a whole is sometimes posted on governmental lists of noxious or quarantine weeds. Our study challenges this stereotype and uses the case of C. jepsonii (Jepson's dodder) to illustrate the precarious biodiversity and conservation status faced by many dodder species. Until now, Jepson's dodder has been known only from its type collection. Consequently, its phylogenetic affinities, morphological variation, and ecology have remained unknown, and the species is currently ambiguously considered either synonymous to the invasive North American weed C. indecora or to an extinct endemic from California. Using molecular data from newly found collections, we infer that C. jepsonii belongs to C. californica species complex, instead of C. indecora clade. Also, we discuss the conservation of this species within the broader biological and ecological context of Cuscuta in general. PMID- 21622361 TI - Microsatellite primers in the white proteas (Protea section Exsertae, Proteaceae), a rapidly radiating lineage. AB - PREMISE OF THE STUDY: Microsatellite primers were developed in the South African sclerophyllous shrub Protea punctata to investigate the degree of population differentiation within and between P. punctata and closely related species. * METHODS AND RESULTS: 10 primer pairs were identified from three individuals of Protea punctata. The primers amplified di- and tri-nucleotide repeats. Across all P. punctata samples, the loci have 8-49 alleles. All primers also amplified in Protea section Exsertae (P. aurea, P. aurea subsp. potbergensis, P. mundii, P. venusta, P. lacticolor, and P. subvestita). The loci had 14-69 alleles across the subgenus. * CONCLUSIONS: These results show the broad utility of microsatellite loci for future studies of population genetics in the white proteas and their potential utility across the entire genus. PMID- 21622362 TI - Female gametophyte and early seed development in Peperomia (Piperaceae). AB - The evolution of female gametophyte development provides an example of how minor ontogenetic modifications can impact the functional biology of seeds. Mature Peperomia-type female gametophytes are normally depicted as 16-nucleate, nine celled structures. However, recent ultrastructural data have demonstrated that many previous reports were incorrect, suggesting that our understanding of the Peperomia-type ontogeny is incomplete. In this investigation, female gametophyte and early seed development is described in Peperomia dolabriformis, P. jamesoniana, and P. hispidula. Nuclear positioning, nuclear division, and vacuole morphology are documented during the syncytial stages of development, and two mature female gametophyte cellular configurations are described. Endosperm ploidy is measured in each species using microspectrofluorometry. We conclude that a 10 celled construction is likely the most common cellular configuration in Peperomia and that a three-celled female gametophyte exists in P. hispidula. We also describe how developmental modifications of wall formation could produce the diverse cellular configurations observed throughout Peperomia. Interestingly, the onset of female gametophyte diversification within Piperales correlates with the origin of the perisperm in the common ancestor of Piperaceae + Saururaceae. We posit that the origin of the perisperm may have relaxed selection on endosperm genetic constructs, thereby promoting diversification of female gametophyte ontogeny. PMID- 21622363 TI - Recalcitrancy and a new kind of epicotyl dormancy in seeds of the understory tropical rainforest tree Humboldtia laurifolia (Fabaceae, Ceasalpinioideae). AB - We report a new kind of seed dormancy and identify the storage behavior category for an important understory rainforest tree that also is used as an ornamental. While studying seed dormancy of Fabaceae species in Sri Lanka, we observed a considerable delay in emergence of the plumule following radicle emergence in Humboldtia laurifolia. Because epicotyl dormancy has not been reported in Fabaceae, we undertook a detailed morphological study of seed germination in this species. Our aims were to document desiccation tolerance/intolerance and epicotyl dormancy in seeds of H. laurifolia. Drying and low temperature storage were used to evaluate storage behavior of the seeds and imbibition, germination, and seed coat anatomy to categorize seed dormancy in two seed collections. Plumule development before its emergence and effects of light and temperature on plumule emergence were monitored. All seeds that were dried to 15% moisture content or stored at -1 degrees C lost viability. Plumules began to grow 20 +/- 5 d from radicle emergence and emerged after 40 +/- 3 d. Dark and high illuminance further delayed plumule emergence. Seeds are recalcitrant and have a hitherto unreported kind of epicotyl dormancy, for which we propose the formula . PMID- 21622364 TI - Ontogenetic changes in the numbers of short- vs. long-shoots account for decreasing specific leaf area in Acer rubrum (Aceraceae) as trees increase in size. AB - Specific leaf area (SLA) is reported to decrease with increasing plant size among dicot tree species despite a strong positive correlation between SLA and relative growth rate. This diminishing returns in SLA may result from changes in the relative numbers of different shoot types bearing leaves with different SLAs as trees increase in overall size. This ontogenetic shift hypothesis was examined for 15 Acer rubrum trees differing in basal stem diameter (0.01 m <= D <= 0.62 m). Detailed analyses of the largest tree showed that short-shoots produced leaves with significantly smaller SLA than the leaves produced by long-shoots regardless of the location of shoots within the canopy. A combination of random effect and split-plot (main-effect) ANOVA models showed that >94% of the variance observed for SLA was attributable to shoot type rather than to the location of leaves in the canopy. Further, with increasing trunk diameter, the number of short-shoots increased rapidly relative to the number of long-shoots. Although the leaves of short-shoots gain disproportionately more surface area per unit mass investment compared to the leaves produced by long-shoots, our data show that ontogenetic shifts occurring at the shoot and whole plant level account for size-dependent decreases in total canopy SLA. PMID- 21622365 TI - Consequences of variation in flowering time within and among individuals of Mertensia fusiformis (Boraginaceae), an early spring wildflower. AB - Climate change is causing many plants to flower earlier in spring, exposing them to novel selection pressures, including-potentially-pollinator shortages. Over 2 years that contrasted in timing of flowering onset, we studied reproductive strategies, pollen limitation, and selection on flowering time in Mertensia fusiformis, a self-incompatible, spring-flowering perennial. Plants opened most of their flowers early in the flowering period, especially in 2007, the early year; but selection favored early-flowering individuals only in 2008. However, resource allocation to early vs. late seed production was flexible: In 2008, but not 2007, early flowers on a plant produced more and heavier seeds. Late flowers were capable of equal seed production if fertilization of early ovules was prevented, suggesting that late flowers serve a bet-hedging function. Evidence for pollen limitation was weak, although there was a tendency for early flowers to be pollen-limited in 2007 and for late flowers to be pollen-limited in 2008. Poor reproductive success in 2007 was likely attributable less to pollen limitation than to frost damage to flowers. We suggest that plasticity in floral longevity and resource allocation among flowers will make this species resilient to short-term pollinator deficits; whether this will help or hinder future adaptation is unclear. PMID- 21622366 TI - The complete chloroplast genome of tall fescue (Lolium arundinaceum; Poaceae) and comparison of whole plastomes from the family Poaceae. AB - In this paper, we describe the complete chloroplast genome of Lolium arundinaceum. This sequence is the culmination of a long-term project completed by >400 undergraduates who took general genetics at Middle Tennessee State University from 2004-2007. It was undertaken in an attempt to introduce these students to an open-ended experiential/exploratory lesson to produce and analyze novel data. The data they produced should provide the necessary information for both phylogenetic comparisons and plastome engineering of tall fescue. The fescue plastome (GenBank FJ466687) is 136048 bp with a typical quadripartite structure and a gene order similar to other grasses; 56% of the plastome is coding region comprised of 75 protein-coding genes, 29 tRNAs, four rRNAs, and one hypothetical coding region (ycf). Comparisons of Poaceae plastomes reveal size differences between the PACC (subfamilies Panicoideae, Arundinoideae, Centothecoideae, and Chloridoideae) and BOP (subfamilies Bambusoideae, Oryzoideae, and Pooideae) clades. Alignment analysis suggests that several potentially conserved large deletions in previously identified intergenic length polymorphic regions are responsible for the majority of the size discrepancy. Phylogenetic analysis using whole plastome data suggests that fescue closely aligns with Lolium perenne. Some unique features as well as phylogenetic branch length calculations, however, suggest that a number of changes have occurred since these species diverged. PMID- 21622367 TI - Fossil Ericaceae from New Zealand: Deconstructing the use of fossil evidence in historical biogeography. AB - The Australasian Ericaceae epitomize many problems in understanding the biogeography of the southern hemisphere, especially the relative contributions of Gondwanan vicariance and dispersal. Late Cretaceous fossil pollen of the family suggests extreme antiquity of the group in Australasia, but recent phylogenetic evidence suggests much younger histories for most of the groups in that region. This paper documents two new species of latest Oligocene-Early Miocene macrofossils of Ericaceae from New Zealand. Cyathodophyllum novae-zelandiae G.J.Jord. & Bannister gen. and sp. nov. is the oldest record of the tribe Styphelieae, but is of a clade now extinct in New Zealand, possibly related to the Tasmanian genus Cyathodes. Richeaphyllum waimumuensis G.J.Jord. & Bannister sp. nov. is a member of Richeeae, but it is ambiguous as to whether it is a member of the impressive modern New Zealand radiation in Dracophyllum. These fossils emphasize the fact that at least some of the fossil pollen of Ericaceae may have been derived from extinct lineages and therefore should not be used as evidence for the antiquity of any modern New Zealand clade of Ericaceae. New fossils and/or detailed analysis of fossil and extant pollen may help resolve such uncertainty. PMID- 21622368 TI - A fruit and leaves of Rhamnaceous affinities from the late Cretaceous (Maastrichtian) of Colombia. AB - Although Rhamnaceae are distributed worldwide today, the former hypothesis for its Laurasian origins may need to be reconsidered in view of recently discovered fossils from the Guaduas Formation of central Colombia (~68 Ma), including fossil leaves (Berhamniphyllum) and a fruit (Archaeopaliurus boyacensis). Recent phylogenetic studies calibrate the stem of Rhamnaceae at 64 Ma and divide the family into three groups, rhamnoid, ampeloziziphoid, and ziziphoid. Although the fruit, Archaeopaliurus boyacensis, resembles that of Paliurus and suggests a relationship to the Paliureae in the ziziphoid group, associated leaves of Berhamniphyllum conform in architecture to those found today only in the Rhamneae tribe. The Maastrichtian age of these fossils predates the ages previously estimated for the Rhamneae tribe (28.5 Ma) and the Paliureae tribe (31.6 Ma) using a modified clock approach. Based on the new megafossil evidence, two alternative ideas are proposed: (1) that these fossils predate the extant tribes or (2) that the family diversified into modern tribes earlier than previously proposed. PMID- 21622369 TI - Early vessel evolution and the diverisification of wood function: Insights from Malagasy Canellales. AB - Xylem vessels have long been proposed as a key innovation for the ecological diversification of angiosperms by providing a breakthrough in hydraulic efficiency to support high rates of photosynthesis and growth. However, recent studies demonstrated that angiosperm woods with structurally "primitive" vessels did not have greater whole stem hydraulic capacities as compared to vesselless angiosperms. As an alternative to the hydraulic superiority hypothesis, the heteroxylly hypothesis proposes that subtle hydraulic efficiencies of primitive vessels over tracheids enabled new directions of functional specialization in the wood. However, the functional properties of early heteroxyllous wood remain unknown. We selected the two species of Canellales from Madagascar to test the heteroxylly hypothesis because Canellaceae (represented by Cinnamosma madagascariensis) produces wood with vessels of an ancestral form, while Winteraceae, the sister clade (represented by Takhtajania perrieri) is vesselless. We found that heteroxylly correlated with increased wood functional diversity related predominantly to biomechanical specialization. However, vessels were not associated with greater stem hydraulic efficiency or increased shoot hydraulic capacity. Our results support the heteroxylly hypothesis and highlight the importance integrating a broader ecological context to understand the evolution of vessels. PMID- 21622370 TI - Gene flow from foreign provenances into local plant populations: Fitness consequences and implications for biodiversity restoration. AB - Long-distance transplantation of seed material as done in restoration programs has raised concerns about the risks associated with the introduction of maladapted genotypes that may hybridize with neighboring native conspecifics and decrease local population fitness (outbreeding depression). We studied the consequences of gene flow from foreign provenances into local populations in the common grassland species Plantago lanceolata (Plantaginaceae). Three generations of intraspecific hybrids (F(1), F(2), and backcross to the local plants) were produced by controlled crossings between local plants and plants from geographically or environmentally distant populations. Their performance was compared to that of within-population crosses in a field experiment. Early growth in some interpopulation hybrids was significantly reduced, and this decrease in performance was higher in progeny of crosses with the local population from a different habitat than with geographically distant populations. At the end of the growing season, most fitness-related traits of the interpopulation hybrids were close to the average of their parents. Crosses with low-performing foreign parents therefore resulted in reduced fitness of the hybrids compared to the local plants and dilution of local adaptation. We conclude that the introduction of maladapted populations from distant or ecologically distinct environments might, at least temporarily, decrease the fitness of neighboring local plants. PMID- 21622371 TI - Multiple spatial scale patterns of genetic diversity in riparian populations of Ainsliaea faurieana (Asteraceae) on Yakushima Island, Japan. AB - Habitat and geographical features of river systems strongly influence gene flow and spatial genetic patterning in riparian plant populations. We investigated the patterns of genetic diversity within and among populations of Ainsliaea faurieana relative to different spatial conditions (along a river, among rivers, and among regions on an island), based on nuclear and chloroplast microsatellite DNA variations. Within an individual river system, we found higher haplotype diversities in downstream populations, and in a Bayesian analysis of recent migration, we detected unidirectional gene movements from upstream to downstream, indicating water-mediated dispersal along the river. Mantel tests detected no isolation-by-distance in genetic variation, suggesting the maintenance of a metapopulation with wide-range seed dispersal by water. Moreover, the observed high level of genetic differentiation, especially in the cpDNA (F(ST) = 0.539), indicated a metapopulation structure with frequent extinction and colonization. On a larger scale, we found high population differentiation and clear genetic structuring among regions, suggesting that gene flow was restricted by geographical features (mountains separating river systems) for relatively long periods. Our findings of genetic structures based on different spatial conditions elucidated patterns and ranges of historical and contemporary gene movement in a plant species that is persistent in extremely disturbed riparian environments. PMID- 21622372 TI - Genetic structure and breeding system of a rare understory herb, Dysosma versipellis (Berberidaceae), from temperate deciduous forests in China. AB - To evaluate the role of Quaternary refugial isolation in allopatric (incipient) speciation of East Asian temperate forest biotas, we analyzed amplified fragment length polymorphisms (AFLPs) and the breeding system in Dysosma versipellis. The study revealed that D. versipellis is mostly self-incompatible, genetically highly subdivided and depauperate at the population level (e.g., Phi(ST) = 0.572/H(E) = 0.083), and characterized by a low pollen-to-seed migration ratio (r ~ 4.0). The latter outcome likely reflects limited pollen flow in a low-seed disperser whose hypothesized "sapromyophilous" flowers undergo scarce, inefficient, and likely specialized cross-pollination by small Anoplodera beetles, rather than carrion flies as assumed previously. In consequence, fruit set in D. versipellis was strongly pollen-limited. Our AFLP data support the hypothesis of a long-standing cessation of gene flow between western and central eastern populations, consistent with previous chloroplast DNA data. This phylogeographic pattern supports the role of the Sichuan Basin as a floristic boundary separating the Sino-Himalayan vs. Sino-Japanese Forest subkingdoms. Our genetic data of D. versipellis also imply that temperate deciduous forest elements to the west and the east of this basin responded differently to Quaternary climate change, which may have triggered or is leading to allopatric (incipient) speciation. PMID- 21622373 TI - Self-sterility in two Cytisus species (Leguminosae, Papilionoideae) due to early acting inbreeding depression. AB - In most angiosperms, the endosperm develops before the embryo, but with harmony between the two structures until final seed formation. In an embryological study, we show that inbreeding depression causes disharmony in development of the two structures in two Leguminosae shrubs, Cytisus multiflorus and C. striatus. Our main objective was to test the causes of self-sterility in the two species by comparing the embryological development of the self seeds with that of cross seeds. In developing selfed seeds of C. multiflorus, the embryo reaches at most the globular stage and never forms mature seeds, while in C. striatus a few mature selfed seeds are formed. In both species, the main cause of abortion of developing selfed seeds is diminished endosperm development (low values of the ratio of endosperm to embryo), which triggers collapse of the endosperm and embryo. The results indicate that self-sterility in C. striatus is postzygotic because of strong, early inbreeding depression, while in C. multiflorus there exists a mixed pre- and postzygotic mechanism; the prezygotic mechanism causes rejection of some self-pollen tubes in the style/ovary, and the early inbreeding depression triggers abortion of fertilized ovules that escaped that action. PMID- 21622374 TI - Stigma closure and re-opening in Oroxylum indicum (Bignoniaceae): Causes and consequences. AB - The study of plant responses to touch, particularly the responses of leaves, stems, and roots, has a long history. By contrast, floral responses are relatively unexplored. Stigma closure is common in the Bignoniaceae, but the factors influencing it are not well understood. We investigated factors influencing stigma closure and reopening and its effects on pollen receipt in seven Oroxylum indicum trees near Hat Yai, Thailand. The effects of pressure, conspecific and heterospecific pollen, and pollen load (the amount of pollen deposited) on stigma behavior were examined in 270 flowers (of the total 430 flowers evaluated in the entire study). Pressure alone resulted in faster closure than did conspecific pollination and faster reopening than did heterospecific pollination. Stigmas never reopened after conspecific pollination. Pollen load had no effect on stigma behavior. Stigmas discriminated between conspecific and foreign pollen; they reopened only after pollination with the latter. A manipulative experiment revealed that stigma closure did not affect the number of conspecific pollen grains received. We also counted pollen tubes in styles that were either hand-supplemented with outcross conspecific pollen or open pollinated. Pollen tube numbers were highest after light pollination (~900 grains), indicating that interference among pollen grains may occur after pollination with very heavy loads (>6000 grains). Possible fitness consequences of these responses are discussed. PMID- 21622375 TI - Multiple causes of seedling rarity in scrub plum, Prunus geniculata (Rosaceae), an endangered shrub of the Florida scrub. AB - Conservation of an imperiled plant often requires an understanding of its reproductive ecology. Scrub plum (Prunus geniculata) is an endangered Florida shrub endemic to pyrogenic xeric uplands. Although plants are long-lived and may flower profusely, particularly after burning, fruit yield is sparse and seedlings are rare. We investigated potential causes of seedling rarity in scrub plum by studying its floral sex and breeding systems, fruit development, and germination ecology. We also developed a flow chart based on two time-since-fire scenarios to illustrate the cumulative impact of these factors on seedling recruitment. Breeding system experiments suggested partial gametophytic self-incompatibility in this functionally andromonoecious species, with inbreeding depression in self compatible individuals. Predispersal seed predation, the major constraint on seedling recruitment, was lower in the first postburn season. In two field experiments, seed germination was <11% irrespective of treatment. These results indicate that seedling recruitment in scrub plum may be limited by multiple factors, including sexual reproductive failure, high predispersal seed predation, and low seed germination. The chance of a bisexual flower producing a seedling ranges from 5 in 1000 to 5 in 100000, depending on the time since fire. Restoration of scrub plum's historical fire regime may mitigate some of these factors. PMID- 21622376 TI - Molecular systematics of the neotropical genus Psiguria (Cucurbitaceae): Implications for phylogeny and species identification. AB - Varying morphological features in many groups of tropical vines confound identification, requiring molecular tools for distinguishing species. Confusion is amplified in Psiguria, a small genus found in Central and South America and the Caribbean, because male and female flowers of these monoecious plants are widely separated by time and position on a branch. We present the first phylogeny of Psiguria utilizing a combination of eight chloroplast intergenic spacers, the internal transcribed spacer (ITS) regions of the nuclear ribosomal DNA repeat, and the intron of the low-copy nuclear gene serine/threonine phosphatase, for a total aligned length of 9456 base pairs. Analyses include multiple accessions of all species in the genus. The data support the monophyly of Psiguria and elucidate several species boundaries. Also presented are Psiguria-specific DNA barcodes, which include the chloroplast regions: ndhC-trnV, rps16-trnQ, rpoB trnC, ndhF-rpl32, and psbZ-trnM. For the first time, systematists, ecologists, and evolutionary biologists will have the tools to confidently identify species of Psiguria with DNA barcodes that may be useful in other genera of Cucurbitaceae. PMID- 21622377 TI - Floral emasculation reveals pollen quality limitation of seed output in Bulbine bulbosa (Asphodelaceae)1. AB - Pollen commonly limits seed output in natural plant populations, but the relative contributions of pollen quantity and quality to pollen limitation remain largely unexplored. Estimates of these contributions are needed to promote better understanding of the ecological and evolutionary consequences of pollen limitation. We assessed pollen limitation and distinguished how pollen quantity and quality contributed to seed output in the predominantly outcrossing herb Bulbine bulbosa. We compared seed output of emasculated and cross-pollinated plants that could not self-pollinate with that of intact plants. Seed output was similar in emasculated and cross-pollinated plants, but it was about 50% less in intact plants. These findings indicate that the quantity of cross-pollen deposited by pollinators did not limit seed output. However, pollinators also deposited self-pollen, which reduced seed output because selfing disabled ovules, rendering them unavailable for cross-fertilization. We conclude that pollen quality rather than quantity can limit seed output under natural conditions, which supports recent proposals for greater consideration of pollen quality in the study of pollen limitation. PMID- 21622378 TI - Isolation and characterization of novel microsatellite markers for Arctium minus (Compositae). AB - PREMISE OF THE STUDY: Microsatellite primers were developed for the invasive plant Arctium minus to investigate the effects of facultative self-pollination and the biannual habit on population genetic structure, as well as the colonization of the Americas by this Eurasian species. * METHODS AND RESULTS: Sixteen di- and trinucleotide microsatellite loci were identified in six populations. The number of alleles per locus ranged from one to 10, observed heterozygosities ranged from 0.000 to 0.897, and the mean value of F(IS) was 0.316. * CONCLUSIONS: These results indicate the utility of these loci in future studies of population genetics in A. minus. PMID- 21622379 TI - Flower and fruit characters in the early-divergent lamiid family Metteniusaceae, with particular reference to the evolution of pseudomonomery. AB - Evaluating the morphological relationships of angiosperm families that still remain unplaced in the current systems of classification is challenging because it requires comparative data across a broad phylogenetic range. The small neotropical family Metteniusaceae was recently placed within the lamiids, as sister to either the enigmatic Oncothecaceae or the clade (Boraginaceae + Gentianales + Lamiales + Solanales + Vahliaceae). We examined the development of two of the primary diagnostic traits of Metteniusaceae, the moniliform anthers and the unilocular gynoecium. The gynoecium is 5-carpellate, and contains two ovules with a massive, vascularized integument. Late sympetaly and unitegmic ovules support placement of Metteniusaceae in the lamiids. The 5-carpellate gynoecium is consistent with a sister-group relationship between Metteniusaceae and Oncothecaceae. The gynoecium of Metteniusaceae is unusual in that it is monosymmetric throughout ontogeny, which indicates pseudomonomery; the five carpel initials are congenitally fused by their margins and form a single locule; the two ovules develop from the two smallest and most poorly developed lateral carpels. Comparisons with other pseudomonomerous taxa allow us to propose division of the complex processes leading to pseudomonomery into eight characters, including carpel number and fusion, gynoecial symmetry, timing of carpel reduction, and number and position of nonfertile carpels. PMID- 21622380 TI - Angiosperm wood structure: Global patterns in vessel anatomy and their relation to wood density and potential conductivity. AB - Woody stems comprise a large biological carbon fraction and determine water transport between roots and leaves; their structure and function can influence both carbon and hydrological cycles. While angiosperm wood anatomy and density determine hydraulic conductivity and mechanical strength, little is known about interrelations across many species. We compiled a global data set comprising two anatomical traits for 3005 woody angiosperms: mean vessel lumen area (A) and number per unit area (N). From these, we calculated vessel lumen fraction (F = AN) and size to number ratio (S = A/N), a new vessel composition index. We examined the extent to which F and S influenced potential sapwood specific stem conductivity (K(S)) and wood density (D; dry mass/fresh volume). F and S varied essentially independently across angiosperms. Variation in K(S) was driven primarily by S, and variation in D was virtually unrelated to F and S. Tissue density outside vessel lumens (D(N)) must predominantly influence D. High S should confer faster K(S) but incur greater freeze-thaw embolism risk. F should also affect K(S), and both F and D(N) should influence mechanical strength, capacitance, and construction costs. Improved theory and quantification are needed to better understand ecological costs and benefits of these three distinct dimensions. PMID- 21622381 TI - Phylogeography and divergence date estimates of a lichen species complex with a disjunct distribution pattern. AB - Disjunct species distributions may result from a combination of geologic events and long-distance dispersal. The foliose lichen species complex Leptogium furfuraceum-L. pseudofurfuraceum has an intercontinental disjunction pattern. Populations of this species complex are found in western North America, southern South America, Africa, and southern Europe. We conducted a phylogenetic study to reconstruct the biogeographic history of this species complex using two ribosomal genes (ITS and LSU) and a protein-coding gene (partial RPB2). Results indicated that the complex comprises four geographically restricted genetic lineages. A sister relationship was found between populations from the same hemispheres, incongruent with previous data derived from morphological characteristics and geographical classification schemes. Incorporating Bayesian ancestral area reconstruction and Bayesian divergence time estimation, we proposed an evolutionary hypothesis for the species complex. The results suggested that processes of biotic expansion via transoceanic dispersal were responsible for the species divergence and distribution patterns observed today. This study also expands the view that cryptic speciation is not a rare phenomenon among fungi and lichens. PMID- 21622382 TI - A role for leaf epidermis in the control of leaf size and the rate and extent of mesophyll cell division. AB - Little is known about the control of leaf size in plants, yet there must be mechanisms by which organ size is measured. Because the control of leaf size extends beyond the action of individual genes or cells, an understanding of the role of leaf cell layers in the determination of leaf size is warranted. Following the construction of graft chimeras composed of small- and large-leaf genotypes of Nicotiana, bilateral leaf blade asymmetry was observed on leaves possessing either a genetically larger or smaller epidermis on one side of the midrib. Although cell size was unaffected by the genotype of the epidermis, the rate and extent of cell division in leaf epidermis altered the rate and extent of cell division in mesophyll and affected leaf size. The data presented neither prove nor disprove whether the mesophyll impacts epidermal cell division but provide the first unequivocal evidence that the extent of cell division in the leaf epidermis alters the extent of cell division in the mesophyll and is a factor regulating blade expansion and ultimate leaf size. PMID- 21622383 TI - Clonal structure and genetic diversity of three desert phreatophytes. AB - The objective of this paper was to assess clone sizes of three perennial desert plant species with AFLP markers and to relate them to clonal and genetic diversity and to hydroecology. The study was carried out at the southern rim of the Taklamakan Desert, where sexual regeneration is only possible shortly after rare flooding events, resulting in rarely established cohorts with subsequent extensive vertical growth and horizontal clonal spread. In this environment, repeated seedling establishment is excluded. We expected decreasing clonal and genetic diversity with increasing clone size and increasing distance to the groundwater table and a common response pattern among all study species. Maximum sizes of Populus euphratica and Alhagi sparsifolia clones were 121 ha and 6.1 ha, respectively, while Tamarix ramosissima clones reached a maximum size of only 38 m(2). In P. euphratica and A. sparsifolia, clonal diversity declined with increasing clone size and increasing distance to the groundwater table, while genetic diversity remained unaffected. Tamarix ramosissima differed from the other species because of a much smaller clonality. Clone size and clonal diversity were found to be good proxy variables for clone age. Despite the considerable age of the clones, genetic diversity is maintained in the populations. PMID- 21622384 TI - Desert wildfire and severe drought diminish survivorship of the long-lived Joshua tree (Yucca brevifolia; Agavaceae). AB - Extreme climate events are transforming plant communities in the desert Southwest of the United States. Abundant precipitation in 1998 associated with El Nino Southern Oscillation (ENSO) stimulated exceptional alien annual plant production in the Mojave Desert that fueled wildfires in 1999. Exacerbated by protracted drought, 80% of the burned Yucca brevifolia, a long-lived arborescent monocot, and 26% of unburned plants died at Joshua Tree National Park by 2004. Many burned plants <1 m tall died immediately, and survival of all but the tallest, oldest plants declined to the same low level by 2004. Postfire sprouting prolonged survival, but only at the wetter, high-elevation sites. During succeeding dry years, herbaceous plants were scarce, and individuals of Thomomys bottae (pocket gopher) gnawed the periderm and hollowed stems of Y. brevifolia causing many of them to topple. Thomomys bottae damage reduced plant survivorship at low elevation, unburned sites and diminished survival of burned plants in all but the driest site, which already had low survival. Accentuated ENSO episodes and more frequent wildfires are expected for the desert Southwest and will likely shift Y. brevifolia population structure toward tall, old adults with fewer opportunities for plant recruitment, thus imperiling the persistence of this unique plant community. PMID- 21622385 TI - Does polyembryony confer a competitive advantage to the invasive perennial vine Vincetoxicum rossicum (Apocynaceae)? AB - Determining which traits may allow some introduced plant species to become invasive in their new environment continues to be a key question in invasion biology. Vincetoxicum rossicum is an invasive, perennial vine colonizing natural and seminatural habitats primarily in the northeastern United States and southeastern Canada. More than half its seeds exhibit polyembryony, a relatively uncommon condition in which a single seed produces multiple seedlings. For evaluating the potential consequences of polyembryony on invasiveness, V. rossicum plants derived from seeds of three embryonic classes-singlets, doublets, and triplets (one, two, and three seedlings per seed, respectively)-were paired in all combinations intraspecifically and with the co-occurring native herbs Solidago canadensis and Asclepias syriaca in a greenhouse study. Vincetoxicum rossicum biomass was 25-55% greater and follicle production 55-100% greater under intraspecific competition compared with interspecific competition. However, within a competitive environment, follicle production varied little. Regardless of competitive environment, V. rossicum originating from seeds with a greater number of embryos typically performed no better than plants arising from seed with fewer embryos (singlets = doublets = triplets)-except intraspecifically where doublets outperformed singlets, and with S. canadensis where triplets outperformed singlets. Our findings suggest that overall performance and fitness of V. rossicum is higher in monocultures than in mixed stands and that its ability to invade new habitats may not be attributable to the production of polyembryonic seeds. PMID- 21622386 TI - Variation of flower opening and closing times in F1 and F2 hybrids of daylily (Hemerocallis fulva; Hemerocallidaceae) and nightlily (H. citrina). AB - In flowering plants, pollination success is strongly dependent on the timing of when flowers start to bloom and when they start to close. To elucidate the genetic mechanism influencing the timing of flower opening and closing, we obtained F1 and F2 hybrids of Hemerocallis fulva (a diurnally blooming species, pollinated by swallowtail butterflies) and H. citrina (a nocturnally blooming species, pollinated by nocturnal hawkmoths) and observed their flowering behavior from blooming to closing with the use of digital cameras. For flower opening times, F1 hybrids were highly variable, and F2 hybrids showed a bimodal distribution of flower opening times with peaks in both the morning and evening. The ratio of morning flowering and evening flowering among F2 hybrids did not deviate from 1:1. For the start to close time, both F1 and F2 hybrids were similar in showing the major peak in the evening. The ratio of evening closing and morning closing among F2 hybrids did not deviate from 3:1. These results suggest that the time of flower opening and the start of closing are regulated by different major genes. PMID- 21622387 TI - Structural, physiological, and stable carbon isotopic evidence that the enigmatic Paleozoic fossil Prototaxites formed from rolled liverwort mats. AB - New structural, nutritional, and stable carbon isotope data may resolve a long standing mystery-the biological affinities of the fossil Prototaxites, the largest organism on land during the Late Silurian to Late Devonian (420-370 Ma). The tree trunk-shaped specimens, of varying dimensions but consistent tubular anatomy, first formed prior to vascular plant dominance. Hence, Prototaxites has been proposed to represent giant algae, fungi, or lichens, despite incompatible biochemical and anatomical observations. Our comparative analyses instead indicate that Prototaxites formed from partially degraded, wind-, gravity-, or water-rolled mats of mixotrophic liverworts having fungal and cyanobacterial associates, much like the modern liverwort genus Marchantia. We propose that the fossil body is largely derived from abundant, highly degradation-resistant, tubular rhizoids of marchantioid liverworts, intermixed with tubular microbial elements. Our concept explains previously puzzling fossil features and is consistent with evidence for liverworts and microbial associates in Ordovician Devonian deposits, extensive ancient and modern marchantioid mats, and modern associations of liverworts with cyanobacteria and diverse types of fungi. Our interpretation indicates that liverworts were important components of Devonian ecosystems, that some macrofossils and microfossils previously attributed to "nematophytes" actually represent remains of ancient liverworts, and that mixotrophy and microbial associations were features of early land plants. PMID- 21622388 TI - Episodic migration of oaks to Iceland: Evidence for a North Atlantic "land bridge" in the latest Miocene. AB - Dating the subsidence history of the North Atlantic Land Bridge (NALB) is crucial for understanding intercontinental disjunctions of northern temperate trees. Traditionally, the NALB has been assumed to have functioned as a corridor for plant migration only during the early Cenozoic, but recent findings of plant fossils and inferences from molecular studies are challenging this view. Here, we report dispersed pollen of Quercus from Late Miocene sediments in Iceland that shows affinities with extant northern hemispheric white oaks and North American red oaks. Older (15 to 10 Ma) sediments do not contain pollen of Quercus suggesting it arrived after that time. Pollen from the 9-8 Ma Hrutagil locality is indistinguishable from morphotypes common among white and red oaks. In contrast, pollen from the 5.5 Ma Selargil locality has a tectum that is at present confined to North American white and red oaks, indicating a second episode of migration to Iceland. These findings suggest that transatlantic migration of temperate plant taxa may not have been limited by vast areas of sea or by cold climates during the Miocene. Furthermore, our results offer a plausible explanation for the remarkably low degree of genetic differentiation between modern disjunct European and North American oaks. PMID- 21622389 TI - Leaf fossils of Banksia (Proteaceae) from New Zealand: An Australian abroad. AB - Fossils can shed new light on plant biogeography and phylogeny. Pinnately lobed leaves from the Oligo-Miocene Newvale lignite mine, South Island, New Zealand are the first extra-Australian leaf fossils of the charismatic genus Banksia (Proteaceae), and they are assigned to a new species, B. novae-zelandiae. Comparison with extant taxa shows that the fossils are best regarded as an extinct stem relative of Banksia because their available features are either plesiomorphic for the genus (notably, the stomata are superficially placed, not sunken in balloon-like pits as in many extant species) or lack evidence of synapomorphies that would enable them to be placed in the crown group. Banksia novae-zelandiae does, however, exhibit two cuticular features that are unique or highly derived for Banksia. These are rugulate subsidiary cell ornamentation and the presence of complex papillae that extensively cover the abaxial leaf surface. The fossils add to the widespread records of the pinnately lobed leaf form in Banksia in Australia beginning in the late Paleocene. This form is now limited to species confined to sclerophyllous heathlands of Mediterranean climate in southwestern Australia. Banksia novae-zelandiae could be part of a lineage that had a long history in New Zealand, perhaps dating to the early Paleogene. PMID- 21622390 TI - Does selfing or outcrossing promote local adaptation? AB - The degree to which plants self-fertilize may impact their potential for genetic adaptation. Given that the mating system influences genetic processes within and among populations, the mating system could limit or promote local adaptation. I conducted a literature survey of published reciprocal transplant experiments in plant populations to quantify the effect of mating system on the magnitude of local adaptation. Mating system had no effect on local adaptation. I detected no effect when species were categorized as either self-compatible or self incompatible or when accounting for environmental differences between source populations. The results suggest that, despite limited genetic variation in selfing species and greater potential for gene flow in outcrossing species, mating system has little influence on adaptation of populations. PMID- 21622391 TI - Genetic effects of chronic habitat fragmentation revisited: Strong genetic structure in a temperate tree, Taxus baccata (Taxaceae), with great dispersal capability. AB - Tree species are thought to be relatively resistant to habitat fragmentation because of their longevity and their aptitude for extensive gene flow, although recent empirical studies have reported negative genetic consequences, in particular after long-term habitat fragmentation in European temperate regions. Yet the response of each species to habitat loss may differ greatly depending on their biological attributes, in particular seed dispersal ability. In this study, we used demographic and molecular data to investigate the genetic consequences of chronic habitat fragmentation in remnant populations of Taxus baccata in the Montseny Mountains, northeast Spain. The age structure of populations revealed demographic bottlenecks and recruitment events associated with exploitation and management practices. We found a strong genetic structure, both at the landscape and within-population levels. We also detected high levels of inbreeding for a strictly outcrossing species. Chronic forest fragmentation resulting from long term exploitation in the Montseny Mountains seems the most plausible explanation for the strong genetic structure observed. Our results support the view that, contrary to some predictions, tree species are not buffered from the adverse effects of habitat fragmentation, even in the case of species with a high dispersal potential. PMID- 21622392 TI - Twin oil sacs facilitate the evolution of a novel type of pollination unit (meranthium) in a South African orchid. AB - The unique floral morphology of the South African orchid H. pulchra, with its twin meranthia, is best explained as an adaptation to pollination by oil collecting bees. Flowers consisting of meranthia (floral parts that function as single pollination units; commonly observed in garden Iris) are extremely rare among the angiosperms and their significance poorly understood. Unlike all other known examples of meranthia, the novel type described for H. pulchra is not bilabiate. All Huttonaea species are unique in having twin petal sacs with glandular verrucae that secrete oil and are pollinated by Rediviva (Melittidae) oil-collecting bees. But only Huttonaea pulchra has long and widely divergent petal claws that place the oil sacs well beyond the reach of a centrally positioned bee. The wide separation of these sacs forces the pollinator, R. colorata, to visit each side of the flower independently and effectively divides the flower into two meranthia. Molecular data indicate that the evolution of the Huttonaea-type meranthium was dependent on the prior evolution of the oil flower/oil bee relationship. Meranthium evolution was also facilitated by the presence of oil in two separate structures (petal sacs) that were not physically constrained to remain in close proximity. PMID- 21622393 TI - Evolution and polyploid origins in North American Arctic Puccinellia (Poaceae) based on nuclear ribosomal spacer and chloroplast DNA sequences. AB - The proportion of polyploid plant species increases at higher latitudes, and it has been suggested that original postglacial Arctic immigrants of some large groups, including grasses, were polyploid. We analyzed noncoding nuclear and chloroplast DNA of all North American diploid Puccinellia (Poaceae) and a subset of arctic polyploids to hypothesize evolutionary relationships among diploids and to evaluate the parentage of polyploids. Diploids formed three lineages: one uniting arctic species P. arctica and P. banksiensis; a second comprising arctic species P. tenella, P. alaskana, P. vahliana, and P. wrightii; and a third uniting the two temperate species P. lemmonii and P. parishii. The arctic species P. angustata (hexaploid) and P. andersonii (primarily octoploid) apparently derive from the P. arctica-P. banksiensis lineage based on ITS and chloroplast sequences, and share an ancestor with arctic triploid/tetraploid P. phryganodes based on nrDNA sequences. Sequence comparisons also suggest tetraploid P. bruggemannii evolved from two arctic lineages: P. vahliana-P. wrightii and P. arctica-P. banksiensis. These patterns and the predominance of arctic rather than temperate diploid species support the idea that diploid Puccinellia recolonized the Arctic from northern glacial refugia like Beringia, and also formed stabilized polyploid hybrids during these refugial events or subsequently during postglacial colonization. PMID- 21622394 TI - Phylogenetics of Puya (Bromeliaceae): Placement, major lineages, and evolution of Chilean species. AB - Puya (Bromeliaceae), a large genus of terrestrial bromeliads found throughout a range of elevations in the Andes and central Chile, is of great systematic, evolutionary, and biogeographical interest. This first molecular phylogenetic study of Puya and related bromeliads employs matK, trnS-trnG, rps16, and PHYC sequences. Chloroplast DNA, nuclear DNA, and combined DNA data all place Puya closest to subfamily Bromelioideae. Nuclear and combined data support Puya as monophyletic, and the two subgenera are nonmonophyletic. All data indicate that the Chilean species of Puya are early diverging within the genus, consistent with Chilean genera as the first-diverging members of subfamily Bromelioideae. Central Chile is identified as a key region for understanding the biogeographical history of Bromeliaceae, as is true with other South American plant groups. A complicated history involving early chloroplast capture and later secondary hybridization and/or introgression is seen in Chilean lineages. These events help explain the occurrence of sterile inflorescence tips, floral color and shape, and leaf indument. The ecological radiation of Puya appears coincident with the final, recent rise of the Andes and subsequent high-elevation habitat diversification. Additionally, geographical distribution, rather than moisture or elevational adaptations, correlates to species relationships. Evolution of CAM photosynthesis has occurred multiple times. PMID- 21622395 TI - Genetic analyses of cell death in maize (Zea mays, Poaceae) leaves reveal a distinct pathway operating in the camouflage1 mutant. AB - Controlled cell death is vital for many physiological processes in plants, such as xylem development, the hypersensitive response (HR), and senescence; however, the pathways governing cell death are incompletely understood. Studies of mutants that display a cell-death phenotype have greatly contributed to our knowledge of how this process is regulated. The maize camouflage1 (cf1) mutant displays the novel phenotype of cell-specific death of bundle sheath (BS) cells in discrete yellow leaf tissues. To investigate the BS cell death in cf1 mutants, we characterized potential underlying factors. Hydrogen peroxide (H(2)O(2)) is known to be involved in many cell-death events in plants, including the HR. However, in vivo staining found no accumulation of H(2)O(2) in cf1 mutant leaves. Additionally, genetic analyses determined that functional chloroplasts are required for cf1 BS cell death. These results demonstrate that cf1 BS cell death occurs via a distinct pathway from that seen in a functionally related maize mutant or in the HR, suggesting that cell death in maize leaves can be caused by multiple mechanisms. PMID- 21622396 TI - Formation and function of a new pollen aperture pattern in angiosperms: The proximal sulcus of Tillandsia leiboldiana (Bromeliaceae). AB - Pollen grains are generally surrounded by an extremely resistant wall interrupted in places by apertures that play a key role in reproduction; pollen tube growth is initiated at these sites. The shift from a proximal to distal aperture location is a striking innovation in seed plant reproduction. Reversals to proximal aperture position have only very rarely been described in angiosperms. The genus Tillandsia belongs to the Bromeliaceae family, and its aperture pattern has been described as distal monosulcate, the most widespread aperture patterns recorded in monocots and basal angiosperms. Here we report developmental and functional elements to demonstrate that the sulcate aperture in Tillandsia leiboldiana is not distal as previously described but proximal. Postmeitotic tetrad observation indicates unambiguously the proximal position of the sulcus, and in vitro germination of pollen grains confirms that the aperture is functional. This is the first report of a sulcate proximal aperture with proximal germination. The observation of microsporogenesis reveals specific features in the patterns of callose thickenings in postmeiotic tetrads. PMID- 21622397 TI - New microsatellite loci for Narcissus papyraceus (Amarillydaceae) and cross amplification in other congeneric species. AB - PREMISE OF THE STUDY: Microsatellite loci from a genomic library of the species Narcissus papyraceus were optimized and characterized for studies of population genetics. * METHODS AND RESULTS: Eleven markers that were successfully amplified showed polymorphism when tested on 50 individuals from two populations in southern Spain and northern Morocco. Overall, the number of alleles per locus ranged between 4 and 15. Between 8 and 11 loci successfully amplified in other eight Narcissus species. * CONCLUSIONS: These markers will enable genetic diversity studies of N. papyraceus across its distribution range and conduct paternity analyses among individuals differing in flower morphology. PMID- 21622398 TI - Development of microsatellite markers for the moss Ptychomitrium gardneri (Ptychomitriaceae). AB - PREMISE OF THE STUDY: Microsatellite markers were developed for Ptychomitrium gardneri to study population genetics of this eastern Asian-North American disjunct moss. * METHODS AND RESULTS: A total of 13 microsatellite markers were developed in Chinese populations of P. gardneri, using the Fast Isolation by AFLP of Sequences COntaining Repeats protocol. Eight of the markers showed polymorphism when assessed in a sample of four populations of 29 individuals from China. These markers amplified three to four alleles per locus. Five primers also amplified in P. linearifolium and P. wilsonii. * CONCLUSIONS: These markers may be useful for further investigation of population genetics of P. gardneri. PMID- 21622399 TI - Characterization of polymorphic microsatellite loci in Haageocereus (Trichocereeae, Cactaceae). AB - PREMISE OF THE STUDY: Microsatellites were isolated from two species of the genus Haageocereus (H. tenuis and H. pseudomelanostele) to be applied in studies of genetic diversity and population structure. * METHODS AND RESULTS: Five loci were employed in a preliminary study of genetic diversity and population differentiation in two rare (H. tenuis and H. repens) and two widespread (H. acranthus and H. pseudomelanostele) species, yielding between one and 44 alleles per locus. All five loci were polymorphic, with overall levels of observed and expected heterozygosities ranging from 0.478 to 0.871 and from 0.564 to 0.956, respectively. Three additional loci were scored in H. pseudomelanostele. These eight plus the remaining 11 loci were amplified from putative parents of three hybrids involving Haageocereus and Espostoa. * CONCLUSIONS: These markers will facilitate analysis of genetic diversity, hybridization, and population differentiation throughout Haageocereus and Espostoa. PMID- 21622400 TI - Microsatellite primers in Rhamnus cathartica (Rhamnaceae) and applicability in related taxa to assess hybridization events. AB - PREMISE OF THE STUDY: Microsatellite primers were developed in the introduced shrub Rhamnus cathartica L., to investigate potential hybridization events with related taxa as a stimulus of invasive spread. * METHODS AND RESULTS: Using a non radioactive protocol, 15 primer sets were identified in North American and European populations of R. cathartica. The primers amplified di-, tri- and pentanucelotide repeats with 1-14 alleles per locus. Most primers also amplified in R. lanceolata, R. smithii, and/or Frangula alnus (previously R. frangula). * CONCLUSIONS: These results indicate the utility of primers in R. cathartica for future genetic studies as well as their potential applicability in other related taxa. PMID- 21622401 TI - Developmental morphology of seedling and shoot and phylogenetic relationship of Diplobryum koyamae (Podostemaceae). AB - We demonstrated that saltational evolution was recurrent in the body plans of seedlings of the aquatic angiosperm Podostemaceae, in contrast to other angiosperms with seedlings having almost common body plans. Diplobryum koyamae, transferred to the genus Hydrodiscus described in this paper, has long-floating shoots with an anchoring disk-like base and is rootless. Such a body plan is distinct from other members of Asian Podostemoideae comprising reduced or moderate shoots borne on the root. Here, our molecular phylogenetic analysis revealed that H. koyamae is sister to a crustose-rooted group of Hanseniella, Hydrobryum and Thawatchaia within Asian Podostemoideae. The germinating embryo was devoid of plumule and radicle, and comprised a single cotyledon and a short hypocotyl, which produced an adventitious shoot endogenously. The leaves are formed in the absence of the shoot apical meristem, accompanying the separation of lightly stained cells. Comparison with other species of Asian Podostemoideae having the plumule and the adventitious root in the seedling, along with their phylogenetic relationship, suggests that saltational evolution occurred in the seedling body plan of H. koyamae leading to the extraordinary adult body plan, as in the separate clade of Dalzellia, Indodalzellia, and Indotristicha of the subfamily Tristichoideae. PMID- 21622402 TI - Allometry and stilt root structure of the neotropical palm Euterpe precatoria (Arecaceae) across sites and successional stages. AB - The lack of secondary meristems shapes allometric relationships of the palms, influencing species segregation according to their capacity to adjust form and function to spatial and temporal changes in environmental conditions. We examined the stem height vs. diameter allometry of Euterpe precatoria across environmental gradients and measured how terrain inclination and palm size affected stilt root structure at two sites in Costa Rica. We dissected the root cone into eight variables and used principal component analysis to summarize their correlation structure. The fit of the stem diameter-height relationship to the stress, elastic, and geometric similarity models was examined using data from 438 palms. Terrain inclination did not affect stilt roots, whose structure was determined by palm size. Palms under 1 m showed geometric similarity, whereas palms above 1 m had slope values that were one and a half times higher, independent of successional stage, and did not adjust to any mechanical model. Taller palms departed from these models when they were large because they had stilt root support. We conclude that height in E. precatoria is constrained by structural support at the base and that diameter at the base of the stem and stilt roots balances height increments over all the size ranges examined. PMID- 21622403 TI - An open-flower mutant of Melilotus alba: Potential for floral-dip transformation of a papilionoid legume with a short life cycle? AB - This paper describes an open-flower mutant, designated opf, that we discovered in a genetic screen of fast neutron bombardment mutants in an attempt at floral-dip transformation of Melilotus alba (Fabaceae; white sweetclover), an alternative papilionoid legume host for Sinorhizobium meliloti. The opf mutant developed flowers with reflexed sepals and petals, thereby exposing the stamens and carpel, whereas wild-type sweetclover inflorescences developed closed flowers where the young stamens and carpel remain covered during the early stages of flower development. Based on crosses with the wild type, the mutant segregated as a single, Mendelian recessive. Crosses were successful only when the opf mutant served as the female parent, suggesting that the mutant was male sterile. However, no obvious differences from wild-type stamen development were observed in the opf mutant. The anther defect was due to indehiscence. However, as the plants approached the end of their life cycle, the frequency of selfing increased. We also investigated whether the opf mutant could be transformed via Agrobacterium tumefaciens floral-dip infiltration because open flowers like those of Arabidopsis appear to be more readily transformable. However, similar to wild type M. alba, the opf mutant is refractory to floral-dip transformation by Agrobacterium tumefaciens. PMID- 21622404 TI - Phenotypic plasticity and precipitation response in Sonoran Desert winter annuals. AB - Temporal environmental variation has profound influences on population dynamics and community structure. Examination of functional traits that influence resource uptake and allocation can illuminate how co-occurring species translate environmental variation into different demographic outcomes, yet few studies have considered interspecific differences in trait plasticity. We experimentally manipulated soil moisture to test the hypothesis that differences in morphological plasticity contribute to species differences in demographic response to unpredictable precipitation in Sonoran Desert winter annual plants. We compared plasticity of leaf traits and biomass allocation between Pectocarya recurvata (Boraginaceae) and Stylocline micropoides (Asteraceae), co-occurring species that differ in long-term demographic patterns. The species with highly variable population dynamics, Stylocline, had striking increases in leaf area and root biomass in response to an experimental increase in soil moisture. In contrast, the species with buffered long-term population dynamics, Pectocarya, did not differ in leaf morphology or biomass allocation between soil moisture treatments. Regardless of water treatment, Pectocarya had earlier reproductive phenology and greater fecundity than Stylocline, suggesting that differences in the timing of the phenological transitions from vegetative to reproductive growth may affect species' responses to precipitation pulses. Combining long-term observations with experimental manipulations provides a window into the functional underpinnings and demographic consequences of trait plasticity. PMID- 21622405 TI - Patterns of genetic diversity in colonizing plant species: Nassauvia lagascae var. lanata (Asteraceae: Mutisieae) on Volcan Lonquimay, Chile. AB - The effect of colonization on the distribution of genetic diversity within and among populations in relation to species characteristics remains an open empirical question. The objective of this study was to contrast genetic diversity within and among established and colonizing populations of Nassauvia lagascae var. lanata on Volcan Lonquimay (Araucania Region, Chile), which erupted on 25 December 1988, and relate genetic diversity to biological characteristics of the populations. We analyzed a total of 240 individuals from 15 populations distributed along the Andes Cordillera using AFLP and obtained a total of 307 AFLP bands, of which 97.7% are polymorphic. Values of population differentiation (F(ST)) did not differ significantly among established and colonizing populations, but colonizing populations did have reduced levels of genetic divergence (as indicated by private and rare bands) and genetic variation (e.g., Shannon index). We conclude that a founder effect through limited numbers of founding propagules derived from nearby source populations has not yet been compensated for by subsequent population growth and migration. Low rates of secondary dispersal via running water, kin-structure within populations, and slow population growth seem to contribute to the slow recovery of genetic diversity. PMID- 21622406 TI - Sclerotia of Typhula ishikariensis biotype B (Typhulaceae) from archaeological sites (4000 to 400 BP) in Hokkaido, northern Japan. AB - Despite their close association with human activities, plant pathogenic fungi have rarely been found in archaeological excavations. We report here that a fungus was closely associated with human activities even in prehistoric times. Sclerotium-like objects were found at historical sites (4000 to 400 BP) on the island of Hokkaido, northern Japan. They were spherical, 0.3-1.0 mm in diameter, and had a medulla and rind. Some had leaf fragments on the surface or a protuberance that resembled emerging sporocarp primordia. These traits indicated that they were sclerotia of the snow mold fungus, Typhula ishikariensis biotype B. PMID- 21622407 TI - Leaf economic traits from fossils support a weedy habit for early angiosperms. AB - Many key aspects of early angiosperms are poorly known, including their ecophysiology and associated habitats. Evidence for fast-growing, weedy angiosperms comes from the Early Cretaceous Potomac Group, where angiosperm fossils, some of them putative herbs, are found in riparian depositional settings. However, inferences of growth rate from sedimentology and growth habit are somewhat indirect; also, the geographic extent of a weedy habit in early angiosperms is poorly constrained. Using a power law between petiole width and leaf mass, we estimated the leaf mass per area (LMA) of species from three Albian (110-105 Ma) fossil floras from North America (Winthrop Formation, Patapsco Formation of the Potomac Group, and the Aspen Shale). All LMAs for angiosperm species are low (<125 g/m(2); mean = 76 g/m(2)) but are high for gymnosperm species (>240 g/m(2); mean = 291 g/m(2)). On the basis of extant relationships between LMA and other leaf economic traits such as photosynthetic rate and leaf lifespan, we conclude that these Early Cretaceous landscapes were populated with weedy angiosperms with short-lived leaves (<12 mo). The unrivalled capacity for fast growth observed today in many angiosperms was in place by no later than the Albian and likely played an important role in their subsequent ecological success. PMID- 21622408 TI - A new Tertiary Ginkgo (Ginkgoaceae) from the Wuyun Formation of Jiayin, Heilongjiang, northeastern China and its paleoenvironmental implications. AB - The living fossil Ginkgo, a relict of a once dominant gymnosperm, existed and flourished early in the Mesozoic, but only a vague outline of its evolutionary history in the Tertiary has been unveiled. Here we describe a new species, Ginkgo jiayinensis sp. nov., from the Wuyun Formation of Jiayin, China, which is another well-established Tertiary species based on leaf fossils besides G. adiantoides. The most remarkable feature of the new species is the amphistomatic leaves, likely representing a distinct evolutionary line of the genus in this time interval. Ginkgo jiayinensis is similar to the co-occurring G. adiantoides and extant G. biloba in the lower cuticle, but the two latter species clearly differ in having hypostomatic leaves. Ginkgo biloba is used as the nearest living equivalent (NLE) species for both G. adiantoides and G. jiayinensis in reconstructing paleo-CO(2), as it has been used for other fossil species with either hypostomatic or amphistomatic leaves. The two Tertiary species are almost identical in stomatal index in abaxial cuticles (8.4 and 8.5, respectively), showing no strong differences in response to atmospheric CO(2). Amphistomatic G. jiayinensis would therefore have used a different ecological strategy from that of G. adiantoides of the same bed. PMID- 21622409 TI - Clonal and spatial genetic structure within populations of a coastal plant, Carex kobomugi (Cyperaceae). AB - Clarification of clonal growth pattern is critical for understanding the population dynamics and reproductive system evolution of clonal plant species. The contribution of clonality to the spatial genetic structure (SGS) within populations is also an important issue. I examined the spatial distribution of genetic variability within two populations of the coastal plant Carex kobomugi using seven microsatellite loci. Genotyping of 226 and 140 ramets within 14 * 40 m and 14 * 34 m plots on two populations revealed 36 and 33 multilocus genotypes, respectively. To quantify the extent of intermingling among clones, for each genet, I calculated the dominance of ramets belonging to a particular genet within a spatial range of the genet. Furthermore, I analyzed spatial distribution of genotypes within 2 * 2 m and 1 * 2 m quadrats using second-order spatial statistics. These analyses indicated that clones are highly intermingled, suggesting a low level of spatial interaction among clones. Spatial autocorrelation analysis of kinship coefficient including all pairs of ramets showed significantly stronger SGS than analysis considering only pairs between different genets. I conclude that clonal propagation largely contributes to SGS at a fine scale. PMID- 21622410 TI - Phylogenetic relationships and natural hybridization among the North American woody bamboos (Poaceae: Bambusoideae: Arundinaria). AB - In spite of the ecological and economic importance of temperate bamboos, relatively little is known about their population biology or evolutionary history. Recently, hybridization has emerged as a potential source of diversity in this group, as well as an underlying cause of taxonomic problems. As part of a broader phylogenetic study of the temperate bamboos, we report the results of an analysis of the North American Arundinaria gigantea species complex, including estimates of genetic variation and molecular evidence of natural hybridization among A. gigantea, A. tecta, and A. appalachiana. The study involved a comparative analysis of amplified fragment length polymorphisms (AFLPs) and chloroplast DNA sequences representing diversity within and among all three species plus individuals with intermediate or unusual morphological characteristics (putative hybrids). Molecular results support the recognition of three species previously defined on the basis of morphology, anatomy, and ecology, with most of the molecular variance accounted for by among-species variation. Molecular evidence also demonstrates that A. tecta and A. appalachiana are sister species, forming a clade that is significantly divergent from A. gigantea. The role of hybridization in the phylogenetic history of Arundinaria is discussed along with implications for the evolution and taxonomy of the temperate woody bamboos. PMID- 21622411 TI - Are spurred cyathia a key innovation? Molecular systematics and trait evolution in the slipper spurges (Pedilanthus clade: Euphorbia, Euphorbiaceae). AB - The study of traits that play a key role in promoting diversification is central to evolutionary biology. Floral nectar spurs are among the few plant traits that correlate with an enhanced rate of diversification, supporting the claim that they are key innovations. Slight changes in spur morphology could confer some degree of premating isolation, explaining why clades with spurs tend to include more species than their spurless close relatives. We explored whether the cyathial nectar spur of the Pedilanthus clade (Euphorbia) may also function as a key innovation. We estimated the phylogeny of the Pedilanthus clade using one plastid (matK) and three nuclear regions (ITS and two G3pdh loci) and used our results and a Yule model of diversification to test the hypothesis that the cyathial spur correlates with an increased diversification rate. We found a lack of statistical support for the key innovation hypothesis unless specific assumptions regarding the phylogeny apply. However, the young age (hence small size) of the group may limit our ability to detect a significant increase in diversification rate. Additionally, our results confirm previous species designations, indicate higher homoplasy in cyathial than in vegetative features, and suggest a possible Central American origin of the group. PMID- 21622412 TI - Biogeography of cedrela (meliaceae, sapindales) in central and South america. AB - Dated phylogenies have helped clarify the complex history of many plant families that today are restricted to the world's tropical forests, but that have Eocene, Oligocene, and Miocene fossils from the northern hemisphere. One such family is the Meliaceae. Here we infer the history of the neotropical Meliaceae genus Cedrela (17 species), the sister clade of which today is restricted to tropical Asia. Sequences from the nuclear ribosomal spacer region and five plastid loci obtained for all ingroup species and relevant outgroups were used to infer species relationships and for molecular-clock dating under two Bayesian relaxed clock models. The clock models differed in their handling of rate autocorrelation and sets of fossil constraints. Results suggest that (1) crown group diversification in Cedrela started in the Oligocene/Early Miocene and intensified in the Late Miocene and Early Pliocene, and (2) Central American Cedrela species do not form a clade, implying reentry into Central America after the closure of the Panamanian Isthmus. At present, Cedrela is distributed in both dry and humid habitats, but morphological features suggest an origin in dry forest under seasonal climates, fitting with Miocene and Pliocene Cedrela fossils from deciduous forests. PMID- 21622413 TI - Measuring wood specific gravity...Correctly. AB - The specific gravity (SG) of wood is a measure of the amount of structural material a tree species allocates to support and strength. In recent years, wood specific gravity, traditionally a forester's variable, has become the domain of ecologists exploring the universality of plant functional traits and conservationists estimating global carbon stocks. While these developments have expanded our knowledge and sample of woods, the methodologies employed to measure wood SG have not received as much scrutiny as SG's ecological importance. Here, we reiterate some of the basic principles and methods for measuring the SG of wood to clarify past practices of foresters and ecologists and to identify some of the prominent errors in recent studies and their consequences. In particular, we identify errors in (1) extracting wood samples that are not representative of tree wood, (2) differentiating wood specific gravity from wood density, (3) drying wood samples at temperatures below 100 degrees C and the resulting moisture content complications, and (4) improperly measuring wood volumes. In addition, we introduce a new experimental technique, using applied calculus, for estimating SG when the form of radial variation is known, a method that significantly reduces the effort required to sample a tree's wood. PMID- 21622414 TI - The role of nodding stems in the goldenrod-gall-fly interaction: A test of the "ducking" hypothesis. AB - Herbivores are among the most pervasive selective forces acting on plants, and the number of plant chemicals that presumably evolved for defense against herbivory is immense. In contrast, biologists are only beginning to appreciate the important roles that architectural traits can play in antiherbivore defense. One putative architectural-resistance trait is the nodding stem apex of some goldenrods (Solidago; Asteraceae). Individuals of S. altissima genets that undergo temporary nodding in the late spring (i.e., "candy-cane" ramets) have been shown to be more resistant than individuals of erect-stemmed genets to certain apex-attacking herbivores. We tested the hypothesis that the greater resistance of candy-cane ramets is accomplished by the ramets' "ducking" from the herbivores. In a greenhouse experiment, nodding candy-cane ramets were significantly more resistant to oviposition by the gall-inducing fly Eurosta solidaginis than were ramets of the same genets that had been experimentally straightened. The straightened candy-cane stems were just as susceptible to ovipositions as were ramets of erect-stemmed genets. Thus, ducking indeed appears to confer a resistance advantage to candy-cane genets of S. altissima. PMID- 21622415 TI - Isolation and characterization of microsatellite markers in the rare clonal plant, Spiraea virginiana (Rosaceae). AB - PREMISE OF THE STUDY: Microsatellite markers were developed in Spiraea virginiana, a federally threatened native shrub found along stream banks, to identify clonal genotypes and measure population genetic variability. * METHODS AND RESULTS: Eleven primer sets were developed using a non-radioactive protocol. These revealed a moderate level of genetic variation, as indicated by the number of alleles per locus (range = 1-4) and an average observed heterozygosity of 0.595. Select loci also amplified successfully in the related species Spiraea japonica. * CONCLUSION: Development of the markers described here is critical for the genetic identification of clonal plants as a first step in demographic analyses, and is necessary for the future conservation of this rare species. Amplification of the markers in S. japonica suggests their potential utility in research regarding this species. PMID- 21622416 TI - Dehydration-induced expression of a 31-kDa dehydrin in Polypodium polypodioides (Polypodiaceae) may enable large, reversible deformation of cell walls. AB - Current and predicted climate changes caused by global warming compel greater understanding of the molecular mechanisms that plants use to survive drought. The desiccation-tolerant fern Polypodium polypodioides exhibits extensive cell wall folding when dried to less than 15% relative water content (RWC) and rapidly (within 24 h) rehydrates when exposed to water and high humidity. A 31-kDa putative dehydrin polypeptide expressed in partially and fully dry tissues detected via western blotting was present only during drying and rapidly dissipated (within 24 h) upon tissue rehydration. Immunostaining indicates the presence of dehydrin near the cell wall of partially and fully dried tissues. Atomic force microscopy of tracheal scalariform perforations indicates that dry vascular tissue does not undergo significant strain. Additionally, environmental scanning electron microscopy reveals differential hydrophilicity between the abaxial and adaxial leaf surfaces as well as large, reversible deformation. The ability to avoid cell wall damage in some desiccation-tolerant species may be partially attributed to cell wall localization of dehydrins enabling reversible, large cell-wall deformation. Thus, the de novo synthesis of dehydrin proteins and potential localization to the cell walls of these desiccation-tolerant species may play a role in avoiding mechanical failure during drought. PMID- 21622417 TI - Eucalyptus regnans (Myrtaceae): A fire-sensitive eucalypt with a resprouter epicormic structure. AB - Determining the location of buds and bud-forming meristems and hence the level of protection from heat is essential to understanding plant response to fire. Most eucalypts resprout readily from the stem (epicormic resprouting) and the base after felling or high intensity fire. In contrast, Eucalyptus regnans is one of the few eastern Australian fire-sensitive, obligate seeder eucalypts. Some authors have suggested that the relatively weak epicormic resprouting is due to a lack of bud-forming structures. Epicormic strands from the bark and outer xylem of three very large trees and two saplings were examined anatomically. Epicormic bud-forming structures were found in all samples examined. The bud-forming capacity consisted of narrow, radially elongated strips of cells of meristematic appearance. These strips were continuous from the outermost secondary xylem through to the outer bark. Bark was relatively thick at the base of the large trees, but remarkably thin above this basal skirt. Eucalyptus regnans was found to possess the apparently fire-adapted epicormic strands previously described in other eucalypts, thus showing its fire-adapted lineage. However, this fire sensitive species apparently directs much of its resources to rapid height-growth rates in younger trees, rather than to vegetative fire survival. PMID- 21622418 TI - Comparative structure and ontogeny of the foliar domatia in three neotropical myrmecophytes. AB - The origin and timing of the appearance of leaf domatia during the ontogeny of plants are important evolutionary traits driving the maintenance of ant-plant associations. In this study conducted in French Guiana on Hirtella physophora, Maieta guianensis, and Tococa guianensis, we focused on the formation and development of leaf domatia having different morphological origins. We modeled the timing of the onset of these domatia, then compared their morpho-anatomical structure. Although the ontogenetic development of the domatia differed between species, they developed very early in the plant's ontogeny so that we did not note differences in the timing of the onset of these domatia. For H. physophora seedlings, a transitional leaf forms before the appearance of fully developed domatia, whereas in M. guianensis and T. guianensis the domatia forms abruptly without transitional leaves. Moreover, in all cases, the morpho-anatomical structure of the domatia differed considerably from the lamina. All three species had similar morpho-anatomical characteristics for the domatia, indicating a convergence in their structural and functional characteristics. This convergence between taxonomically distant plant species bearing domatia having different morphological origins could be interpreted as a product of the plant's evolution toward the morphology and anatomy most likely to maximize ant recruitment and long-term residence. PMID- 21622419 TI - A phylogenetic circumscription of Polytrichastrum (Polytrichaceae): Reassessment of sporophyte morphology supports molecular phylogeny. AB - Mosses arguably possess the most structurally complex sporangia of any extant land plants, a consequence of being the monosporangiophyte lineage most strongly adapted to terrestrial environments. Morphological and functional variation in the mechanisms that regulate spore release in one of the major classes of mosses, the Polytrichopsida, is largely unexplored, while recent research indicates that the most distinctive structure, the peristome, has evolved independently in the Polytrichopsida and in other mosses. The genus Polytrichastrum was separated from Polytrichum on the basis of such sporangial characters, although the critical features had until recently only been examined using light microscopy, and strong evidence from molecular data indicated that Polytrichastrum as currently circumscribed is polyphyletic. Here we use Bayesian ancestral character state reconstruction in conjunction with extensive scanning electron micrographic studies to elucidate probable morphology at ancestral nodes and define natural taxa. As well as clarifying the structure, evolution, and aspects of development of the peristome-epiphragm complex in this highly prominent group of mosses, the results provide a basis for a revised phylogenetic taxonomy in which the species of Polytrichastrum sect. Aporotheca are recognized once more within Polytrichum. PMID- 21622420 TI - Remototrachyna, a newly recognized tropical lineage of lichens in the Hypotrachyna clade (Parmeliaceae, Ascomycota), originated in the Indian subcontinent. AB - Biogeographical studies of lichens used to be complicated because of the large distribution ranges of many species. Molecular systematics has revitalized lichen biogeography by improving species delimitation and providing better information about species range limitations. This study focuses on the major clade of tropical parmelioid lichens, which share a chemical feature, the presence of isolichenan in the cell wall, and a morphological feature, microscopic pores in the uppermost layer. Our previous phylogenetic studies revealed that the largest genus in this clade, Hypotrachyna, is polyphyletic with a clade mainly distributed in South and East Asia clustering distant from the core of the genus. To divide the Hypotrachyna clade into monophyletic groups and to reevaluate morphological and chemical characters in a phylogenetic context, we sampled ITS, nuclear large subunit (nuLSU) and mitochondrial small subunit (mtSSU) rDNA sequences from 77 species. We are erecting the new genus Remototrachyna for a core group of 15 former Hypotrachyna species. The segregation of Remototrachyna from Hypotrachyna receives support from morphological and chemical data, as well from maximum parsimony, maximum likelihood, and Bayesian phylogenetic analyses of the DNA. We used a likelihood approach to study the geographic range evolution of Remototrachyna and Bulbothrix, which are sister groups. This analysis suggests that the ancestral range of Remototrachyna was restricted to India and that subsequent long-distance dispersal is responsible for the pantropical occurrence of two species of Remototrachyna. PMID- 21622421 TI - Expression patterns of AaDL, a CRABS CLAW ortholog in Asparagus asparagoides (Asparagaceae), demonstrate a stepwise evolution of CRC/DL subfamily of YABBY genes. AB - Understanding the evolutionary path of transcription factors is essential for the elucidation of plant evolution. The CRC/DL subfamily of the YABBY gene family are functionally diverse, plant-specific, putative transcription factors. In Arabidopsis thaliana, CRABS CLAW (CRC) is expressed in the abaxial region of carpel primordia and in floral nectaries, where it regulates carpel morphology and nectary development. By contrast, in Oryza sativa, DROOPING LEAF (DL) is expressed in the entire carpel primordium and in the central undifferentiated cells of leaves, where it regulates carpel identity and midrib development. Recent studies suggest that abaxial expression and functional roles in the carpel are ancestral characters, although when and how neo-functionalizations occurred remains unclear. To elucidate the evolutionary processes of the CRC/DL subfamily, we examined in situ expression patterns of a CRC ortholog (AaDL) in Asparagus asparagoides (Asparagales). Like CRC in Arabidopsis thaliana, AaDL was clearly expressed in the abaxial region of the ovary wall. Expression was also detected in the phloem of leaves, but not in the septal nectary. Thus, expression in the entire carpel primordium might have been acquired after the divergence of Asparagus, with expression competence in the leaves acquired before the divergence of Asparagus in monocots. Our data indicate that the evolution of CRC/DL subfamily genes occurred in a stepwise manner. PMID- 21622422 TI - Within-plant distribution of phenolic glycosides and extrafloral nectaries in trembling aspen (Populus tremuloides; Salicaceae). AB - Expression of foliar secondary compounds and extrafloral nectaries (EFNs) within the same leaves may be incompatible if secondary compounds repel beneficial insects that might otherwise be attracted to EFNs. This study examined the within plant distributions of phenolic glycosides and EFNs in trembling aspen, Populus tremuloides, and their relationships to herbivore damage. Populus tremuloides expresses extrafloral nectaries (EFNs) on a subset of its leaves. We studied short and tall naturally occurring aspen ramets across multiple sites in interior Alaska. Contrary to our expectations, foliar phenolic glycoside concentrations were approximately 30% greater on leaves bearing EFNs than on leaves without EFNs. The mean concentration of foliar phenolic glycosides in short ramets was nine times that in tall ramets. Phenolic glycoside concentration was negatively related to leaf mining damage by Phyllocnistis populiella (Lepidoptera; Gracilliadae) at concentrations greater than 27 mg/g, whereas the presence of EFNs was unrelated to mining damage. The positive association of chemical defensive compounds and EFNs in leaves suggests that, for species with variation in EFN expression, negative correlations between herbivory and EFN expression may arise indirectly from associated effects of other, correlated types of defense. PMID- 21622423 TI - Allometry of within-fruit reproductive allocation in subtropical dicot woody species. AB - Angiosperm fruits typically consist of pericarp and seed, which collectively function to maximize plant reproductive success. Within-fruit reproductive allocation has been scarcely examined across a wide range of fruit types and taxa although it is critical to the understanding of the evolution of fruit size and seed size. We investigated seed size, fruit size, seed number per fruit (SNF), and within-fruit biomass allocation between seed mass and pericarp mass for 62 dicot woody species (27 deciduous and 35 evergreen species) of a subtropical evergreen forest in southwest China. At the fruit level, total pericarp mass (TPM) isometrically scaled with increasing total seed mass (TSM) in the evergreen species and in the pooled data set, while TPM increased faster than TSM in the deciduous species. The slope difference is possibly due to the difference in the timing of fruit development between the two species groups. At the seed level, seed package (pericarp mass per seed) isometrically scaled with increasing seed size in the deciduous group, but less than isometrically in the evergreens and in the pooled data set. SNF was negatively correlated with seed size but positively correlated with the proportion of pericarp within fruits. In conclusion, within fruit biomass allocation is significantly affected by seed size, fruit size, and SNF in both deciduous and evergreen species. The implications of the observed scaling relationships are discussed in relation to seed size evolution and global patterns of seed size variation. PMID- 21622424 TI - The chloroplast genome of Anomochloa marantoidea (Anomochlooideae; Poaceae) comprises a mixture of grass-like and unique features. AB - Features in the complete plastome of Anomochloa marantoidea (Poaceae) were investigated. This species is one of four of Anomochlooideae, the crown node of which diverged before those of any other grass subfamily. The plastome was sequenced from overlapping amplicons using previously designed primers. The plastome of A. marantoidea is 138412 bp long with a typical gene content for Poaceae. Five regions were examined in detail because of prior surveys that identified structural alterations among graminoid Poales. Anomochloa marantoidea was found to have an intron in rpoC1, unlike other Poaceae. The insertion region of rpoC2 is unusually short in A. marantoidea compared with those of other grasses, but with atypically long subrepeats. Both ycf1 and ycf2 are nonfunctional as is typical in grasses, but A. marantoidea has a uniquely long psiycf1. Finally, the rbcL-psaI spacer in A. marantoidea is atypically short with no evidence of the psirpl23 locus found in all other Poaceae. Some of these features are of noteworthy dissimilarity between A. marantoidea and those crown grasses for which entire plastomes have been sequenced. Complete plastome sequences of other Anomochlooideae and outgroups will further advance our understanding of the evolutionary events in the plastome that accompanied graminoid diversification. PMID- 21622425 TI - Rangewide analysis of fungal associations in the fully mycoheterotrophic Corallorhiza striata complex (Orchidaceae) reveals extreme specificity on ectomycorrhizal Tomentella (Thelephoraceae) across North America. AB - Fully mycoheterotrophic plants offer a fascinating system for studying phylogenetic associations and dynamics of symbiotic specificity between hosts and parasites. These plants frequently parasitize mutualistic mycorrhizal symbioses between fungi and trees. Corallorhiza striata is a fully mycoheterotrophic, North American orchid distributed from Mexico to Canada, but the full extent of its fungal associations and specificity is unknown. Plastid DNA (orchids) and ITS (fungi) were sequenced for 107 individuals from 42 populations across North America to identify C. striata mycobionts and test hypotheses on fungal host specificity. Four largely allopatric orchid plastid clades were recovered, and all fungal sequences were most similar to ectomycorrhizal Tomentella (Thelephoraceae), nearly all to T. fuscocinerea. Orchid-fungal gene trees were incongruent but nonindependent; orchid clades associated with divergent sets of fungi, with a clade of Californian orchids subspecialized toward a narrow Tomentella fuscocinerea clade. Both geography and orchid clades were important determinants of fungal association, following a geographic mosaic model of specificity on Tomentella fungi. These findings corroborate patterns described in other fully mycoheterotrophic orchids and monotropes, represent one of the most extensive plant-fungal genetic investigations of fully mycoheterotrophic plants, and have conservation implications for the >400 plant species engaging in this trophic strategy worldwide. PMID- 21622426 TI - Loss of psbS expression reduces vegetative growth, reproductive output, and light limited, but not light-saturated, photosynthesis in Arabidopsis thaliana (Brassicaceae) grown in temperate light environments. AB - Plants protect themselves against the deleterious effects of high light intensities by inducing a mechanism ubiquitous among plants known as energy dissipation, which safely converts excess light to heat before it can lead to the formation of free radicals. Mutants possessing a deletion of the psbS gene, such as the npq4 mutant, cannot perform energy dissipation and thus offer an opportunity to assess the importance of this process to plant function. In a temperate light environment, greenhouse-grown npq4 mutants of Arabidopsis thaliana had smaller rosette diameters and leaf numbers. The reduction in size observed in npq4 plants was associated with fewer floral stalks, fewer fruits, lower whole-plant and individual seed masses, and lower germination rates. In the field, npq4 mutants developed fewer fruits. After a controlled exposure to high light stress, both PSII efficiency and CO(2) assimilation were more significantly compromised in npq4 mutants at low light intensities, but not at high light intensities. Thus, the protective nature of energy dissipation manifests in light environments that include periods of high light, which predispose plants to PSII photoinactivation, and periods of low light, when PSII photoinactivation decreases the rate of photosynthesis. PMID- 21622427 TI - Effects of historical demography and ecological context on spatial patterns of genetic diversity within foxtail pine (Pinus balfouriana; Pinaceae) stands located in the Klamath Mountains, California. AB - The density and dispersion of individuals, nonequilibrium demographics, and habitat fragmentation all affect the magnitude and extent of spatial genetic structure within forest tree populations. Here, we investigate the link between historical demography and spatial genetic structure within ecologically contrasting stands of foxtail pine (Pinus balfouriana) in the Klamath Mountains of northern California. We defined two stand types a priori, based largely on differences in foxtail pine density and basal area, and for each type we sampled two stands. Population expansions, likely from Pleistocene bottlenecks, were detected in three of the four stands. The magnitude and extent of spatial autocorrelation among genotypes at five nuclear microsatellites differed dramatically among stands, with those having lower foxtail pine density exhibiting strong patterns of isolation by distance. Moran's I statistics were 7 fold higher for the first distance class (<25 m) in these stands relative to those observed in stands with higher foxtail pine density (I(25) = 0.14 vs. 0.02). We conclude that differences in spatial genetic structure between stand types are due to differences in ecological attributes that affected expansion from inferred bottlenecks. PMID- 21622428 TI - Genetic variation in walnuts (Juglans regia and J. sigillata; Juglandaceae): Species distinctions, human impacts, and the conservation of agrobiodiversity in Yunnan, China. AB - Walnuts are a major crop of many countries and mostly cultivated in large-scale plantations with few cultivars. Landraces provide important genetic reservoirs; thus, understanding factors influencing the geographic distribution of genetic variation in crop resources is a fundamental goal of agrobiodiversity conservation. Here, we investigated the role of human settlements and kinship on genetic variation and population structure of two walnut species: Juglans regia, an introduced species widely cultivated for its nuts, and J. sigillata, a native species cultivated locally in Yunnan. The objectives of this study were to characterize sympatric populations of J. regia and J. sigillata using 14 molecular markers and evaluate the role of Tibetan villages and kin groups (related households) on genotypic variation and population structure of J. regia and J. sigillata. Our results based on 220 walnut trees from six Tibetan villages show that although J. regia and J. sigillata are morphologically distinct, the two species are indistinguishable based on microsatellite data. Despite the lack of interspecific differences, AMOVAs partitioned among villages (5.41%, P = 0.0068) and kin groups within villages (3.34%, P = 0.0068) showed significant genetic variation. These findings suggest that village environments and familial relationships are factors contributing to the geographic structure of genetic variation in Tibetan walnuts. PMID- 21622429 TI - A reassessment of the function of floral nectar in Croton suberosus (Euphorbiaceae): A reward for plant defenders and pollinators. AB - Typically, plant-pollinator interactions are recognized as mutualistic relationships. Flower visitors, however, can potentially play multiple roles. The floral nectar in Croton suberosus has been proposed to operate as a reward for predators, especially the wasp Polistes instabilis (Vespidae), which kills herbivorous insects, while the plant has been thought to be mainly wind pollinated. In this study, we reassessed the pollination mode of C. suberosus and the possible role of its flower visitors. Pollinator exclusion experiments demonstrated that C. suberosus should be considered a strictly entomophilous species. Inflorescences of C. suberosus were visited by a diverse entomofauna involving 28 taxa belonging to six orders; however, wasps and bees were the only visitors that carried C. suberosus pollen. The visitation rate of wasps was approximately four times that of bees. This observation, combined with the fact that the small size of bees makes effective contact of their bodies with the stigma difficult, strongly suggests that large wasps are responsible for most of the effective pollination of C. suberosus. Among the wasp visitors, P. instabilis seems to be one of the most important. These findings expose an unusual plant insect interaction, in which the plant provides nectar and wasps pollinate and defend the plant. PMID- 21622430 TI - Ecological and geographic modes of species divergence in wild tomatoes. AB - Understanding the role of geography and ecology in species divergence is central to the study of evolutionary diversification. We used climatic, geographic, and biological data from nine wild Andean tomato species to describe each species' ecological niche and to evaluate the likely ecological and geographical modes of speciation in this clade. Using data from >1000 wild accessions and publicly available data derived from geographic information systems for various environmental variables, we found most species pairs were significantly differentiated for one or more environmental variables. By comparing species' predicted niches generated by species distribution modeling (SDM), we found significant niche differentiation among three of four sister-species pairs, suggesting ecological divergence is consistently associated with recent divergence. In comparison, based on age-range correlation (ARC) analysis, there was no evidence for a predominant geographical (allopatric vs. sympatric) context for speciation in this group. Overall, our results suggest an important role for environmentally mediated differentiation, rather than simply geographical isolation, in species divergence. PMID- 21622431 TI - Density of intraspecific competitors determines the occurrence and benefits of accelerated germination. AB - Germination is a key process in plant recruitment and population dynamics, and seeds are expected to be under strong selection pressure to germinate under conditions that maximize subsequent plant survival. Increased rates of germination (i.e., accelerated germination) may occur in competitive environments. We examined the effects of conspecific density on the timing of germination of seeds of a bird-dispersed plant, Phytolacca americana (Phytolaccaceae, L.), in three different competitive environments. By comparing germination of seeds sown at the same time at different densities, we quantify the benefits of accelerated germination under conditions in which differences in performance among seedlings are attributable to germination timing only, and not to being sown at different times. We find that although the probability of germination is unchanged, the time to initiation of germination is significantly shorter when competition is greater. We also show that plants that germinate earlier are larger and have higher growth rates because they have more time to grow without competitors. Our work demonstrates that shifts in germination timing in response to competition can yield significant dividends for seeds that germinate earliest, but we caution that the magnitude and consequences of accelerated germination will likely depend on the competitive neighborhood. PMID- 21622432 TI - Isolation of microsatellite markers for the common Mediterranean shrub Myrtus communis (Myrtaceae). AB - PREMISE OF THE STUDY: The development of microsatellite markers was conducted in the Mediterranean common shrub Myrtus communis (myrtle) to assess levels of genetic diversity and patterns of gene flow across fragmented landscapes in southern Spain. METHODS AND RESULTS: Fourteen primer pairs were isolated showing clear and consistent patterns of amplification, three of which were apparently monomorphic. Levels of polymorphism in the other 11 markers were checked in 48 individuals from two populations. The number of alleles per locus ranged from 3 to 11 and the total number of alleles was 83. CONCLUSIONS: These highly polymorphic markers will allow us to improve our understanding of the genetic consequences of chronic fragmentation in Mediterranean landscapes. PMID- 21622433 TI - Isolation and characterization of 20 microsatellite loci for laurel species (Laurus, Lauraceae). AB - PREMISE OF THE STUDY: Microsatellite primers were developed for the evergreen tree Laurus to investigate population genetic structure and patterns of gene flow via animal-dispersed pollen and seeds. METHODS AND RESULTS: Twenty polymorphic nuclear microsatellite markers were developed using CA, GA, AAC, and ATG n enriched genomic libraries. Given the tetraploidy of the sampled populations, we analyzed our data both as dominant loci and as codominant genotypic data to calculate allele frequencies and genetic diversity. A total of 196 and 222 alleles were found in 37 Mediterranean (L. nobilis) and 26 Macaronesian islands (L. azorica) individuals, respectively. CONCLUSIONS: Levels of polymorphism of the reported markers are adequate for studies of diversity and parentage in natural populations of this Tertiary relict tree. PMID- 21622434 TI - Development and multiplexing of microsatellite markers in the polyploid perennial herb, Menyanthes trifoliata (Menyanthaceae). AB - PREMISE OF THE STUDY: We developed microsatellite primers to investigate genetic diversity and population genetic structure of the endangered herb Menyanthes trifoliata. METHODS AND RESULTS: Using the microsatellite-enriched library method, we identified 10 primer pairs in M. trifoliata. The primers amplified nine di- and one tri-nucleotide repeats with 4-13 alleles per locus in two Belgian populations. CONCLUSIONS: The results indicate that these markers offer an appropriate amount of variation to investigate genetic diversity, pollen dispersal (through paternity inference), and other conservation issues. PMID- 21622435 TI - Characterization of nuclear microsatellite loci in the Neotropical tree Parkia panurensis (Fabaceae). AB - PREMISE OF THE STUDY: We present here a set of nine polymorphic nuclear microsatellite loci, identified for the first time within the neotropical legume tree species Parkia panurensis Benth. ex H. C. Hopkins, which is widespread in western and central Amazonia. METHODS AND RESULTS: To characterize these loci, 33 Parkia panurensis adult trees were analyzed. The number of alleles ranged from eight to 32, with an average of 14.4 alleles per locus. Mean expected heterozygosity ranged from 0.74 to 0.955. CONCLUSIONS: All nine loci could also be verified in six other Parkia species and polymorphic fragments amplified. The new marker set can be used for future studies of genetic diversity and differentiation, as well as estimation of gene flow and parentage analyses in various Parkia species. PMID- 21622436 TI - Isolation and characterization of microsatellite loci in the common milkweed, Asclepias syriaca (Apocynaceae). AB - PREMISE OF THE STUDY: Microsatellite primers were developed for the common milkweed, Asclepias syriaca L., to assist in genet identification and the analysis of spatial genetic structure. METHODS AND RESULTS: Using an enrichment cloning protocol, eight microsatellite loci were isolated and characterized in a Michigan population of A. syriaca. The primers amplified di- and trinucleotide repeats with 4-13 alleles per locus. CONCLUSIONS: The primers will be useful for studies of clonality and gene flow in natural populations. PMID- 21622437 TI - "A case to which no parallel exists": The influence of Darwin's Different Forms of Flowers. AB - PREMISE OF THE STUDY: Research on the subject of heterostyly is often traced back to 1877 when Charles Darwin published the landmark book The Different Forms of Flowers on Plants of the Same Species. This book synthesized heterostyly research at the time, much of which Darwin conducted, and it continues to be a major contribution to the study of the breeding system. In this book, Darwin discussed the ecology, morph-specific differences, self- and intramorph-incompatibility, evolution and origin, and floral development of heterostyly. Many of the hypotheses he proposed have been and continue to be tested. KEY RESULTS: Throughout the 20(th) and 21(st) centuries, researchers have continued to identify new and different morph-specific floral characters, discover the mechanisms that underlie heteromorphic self-incompatibility, use phylogenies to examine the evolution of heterostyly, and determine novel floral developmental patterns in heterostylous species. From all of these studies, we have learned a great deal about the function, evolution, and development of heterostyly. CONCLUSIONS: However, almost 150 years after Darwin's publications on the subject of heterostyly, we still have a great deal to learn concerning the breeding system, and new technologies and techniques are allowing for new advances in heterostyly research to occur. PMID- 21622438 TI - Understanding spikelet orientation in Paniceae (Poaceae). AB - Spikelet structure and grouping are key characters to identify grasses. Here we tested the possibility that spikelet pairs, a distinctive morphological structure of many Andropogoneae and Paniceae, are the starting point for a secondary single spikelet condition that can also explain the change of spikelet orientation among Paniceae genera. As a first approach, we studied the inflorescence development of Paspalum simplex, P. stellatum, and Axonopus sufultus to clarify the origin of the spikelet orientation and other basic homologies. The results support that solitary spikelets of A. suffultus are homologous to the subsessile spikelets of P. simplex and that solitary spikelets of P. stellatum are homologous to the pedicellate spikelet of P. simplex. This last homology supports that spikelet orientation results from a differential reduction/abortion of either the pedicellate or the subsessile spikelet primordia. We also discuss the possibility that the RAMOSA and polar auxin pathways could play a role in the abortion of the lateral subsessile spikelets in P. stellatum. However, the apical meristem inhibition observed in A. suffultus and P. stellatum seems to depend on a very different genetic control, suggesting that the single spikelet condition is homoplasic within Paniceae and derived from at least two different genetic mechanisms. PMID- 21622439 TI - Anatomical relations among endophytic holoparasitic angiosperms, autotrophic host plants and mycorrhizal fungi: A novel tripartite interaction. AB - Mycorrhizae are widespread mutualistic symbioses crucial for the functioning of terrestrial ecosystems. Not all plants associate with mycorrhizae; most parasitic plants have been suggested to be nonmycorrhizal because they have developed alternative strategies to obtain nutrients. In endophytic parasitic plants, whose vegetative bodies grow completely inside their mycorrhizal host roots, the opportunity for establishing a tripartite association seems evident, but information on these systems is lacking. In studying natural associations among the endophytic holoparasite Cytinus hypocistis, their Cistaceae host species, and associated mycorrhizal fungi, we found that mycorrhizae were associated with the hosts and the parasites, reaching high frequencies of colonization. In parasitic and host root tissues, mycorrhizal fungi spread in the parenchymatic cells by intracellular growth and formed hyphal coils and vesicles, while the cambium and the vascular tissues were never colonized. This report is the first on a tripartite association of an endophytic parasitic plant, its host, and mycorrhizae in natural conditions, representing a novel trophic interaction not previously reported within the angiosperms. Additional studies on the interactions occurring among these three players are needed because they may be crucial to our understanding of how this mutualistic-antagonistic system is functioning and evolving. PMID- 21622440 TI - Symbiotic lifestyle and phylogenetic relationships of the bionts of Mastodia tessellata (Ascomycota, incertae sedis). AB - The biological nature of some symbioses is unclear because it is often not easy to discern whether the symbionts obtain any benefits from the association. Mastodia tessellata, a symbiosis between a leafy green alga and a fungus of uncertain phylogenetic position, is among the most investigated, controversial, and poorly understood associations. Because it has been difficult to determine whether this association is mutually beneficial or parasitic, not all scientists accept M. tessellata as a true lichen symbiosis. Mastodia tessellata is thus an interesting model to illustrate the interactions and processes that occur in fungal-algal symbioses. To improve our understanding of this association, we address the phylogenetic positions of the bionts involved and examine their interactions at the ultrastructural level. Examining the nuLSU and nuSSU gene regions of the mycobiont and the rbcL gene region of the photobiont, we found the fungus to be related to a group of marine species in the genus Verrucaria, family Verrucariaceae, despite its present ascription to the family Mastodiaceae. In addition, the photobiont of the symbiosis emerged as closely related to the North American species Prasiola borealis. Our electron microscopy observations provide new information on the process of fungal colonization of the algal thalli, as well as on relationships between the symbionts during different stages of colonization. The special features of this lichen symbiosis are discussed and compared with other examples of fungal symbioses in nature. PMID- 21622441 TI - Drought stress signals in modern and subfossil Quercus laurifolia (Fagaceae) leaves reflect winter precipitation in southern Florida tied to El Nino-Southern Oscillation activity. AB - In the present study, structural xeromorphic features in modern and subfossil Quercus laurifolia leaves from southern Florida were quantified to reconstruct past precipitation changes in sensitive terrestrial settings. Absolute cell numbers/mm(2), quantified as epidermal cell density (ED) have been analyzed on leaves from herbarium collections as well as the leaves accumulated during the past 125 years in peat deposits. The results reveal a common principal correlation between the measured ED and winter precipitation (November through March, NDJFM: Herbarium r = -0.74; peat profiles FAK98 r = -0.72, FAK02 r = 0.53) providing a measure of seasonal drought stress. In Florida, the amount of winter precipitation depends on El Nino-Southern Oscillation (ENSO) activity, where El Nino years produce wet and cold winters, while La Nina winters are dry and warm. The negative correlation between cell numbers and winter precipitation has the potential to record precipitation variability from subfossil leaves on near-annual to decadal time scales. In subtropical, terrestrial environments, where traditional paleo-proxies are limited, systematic analysis of leaf morphological characteristics can provide important information on precipitation changes through time. PMID- 21622442 TI - Population dependence in the interactions with neighbors for pollination: A field experiment with Taraxacum officinale. AB - PREMISE OF THE STUDY. The fitness of plants depends on their immediate biotic and abiotic environmental surroundings. The floral neighborhood of individual plants is part of this immediate environment and affects the frequency and behavior of their pollinators. However, the interactions among plants for pollination might differ among populations because populations differ in floral densities and pollinator assemblages. Despite that, manipulative experiments of the floral neighborhood in different populations with a specific focus on pollinator behavior are still rare. METHODS. We introduced mixtures of two species (Salvia farinacae and Tagetes bonanza) in two populations of Taraxacum officinale and examined their effect on pollinators' foraging behavior on Taraxacum. KEY RESULTS. The effects of the heterospecific neighborhood differed among pollinator groups and between the two populations. Only honeybees consistently preferred both the most diverse (containing three species) and completely pure patches of Taraxacum in both populations. We found a strong and positive effect of patch diversity on visitation to Taraxacum in one population, whereas in the other population either no effect or a negative effect of plant diversity occurred, which we attribute to differences between populations in the ratio of pollinators to inflorescences. Pollinator visitation consistently increased with local Taraxacum density in both populations. CONCLUSIONS. Our study shows that a similar local neighborhood can differentially affect the frequency and foraging behavior of pollinators, even in closely situated populations. Experimental studies conducted in several populations would contribute to determine which factors drive the variation in pollination interactions among populations. PMID- 21622443 TI - Population dynamics of Empetrum hermaphroditum (Ericaceae) on a subarctic sand dune: Evidence of rapid colonization through efficient sexual reproduction. AB - The importance of sexual reproduction for clonal plant species has long been underestimated, perhaps as a consequence of the difficulty in identifying individuals, preventing the study of their population dynamics. Such is the case for Empetrum hermaphroditum, an ericaceous species, which dominates the ground vegetation of subarctic ecosystems. Despite abundant seed production, seedlings are rarely observed. Therefore, prevalent seedling recruitment on a subarctic dune system provided an opportunity to study the population dynamics and spatial pattern of the colonization phase of this species. We established a 6-ha grid on the dune systems that extended from the shoreline to the fixed dunes and mapped and measured all E. hermaphroditum individuals in the grid. Moreover, we sampled 112 individuals just outside the grid to identify any allometric relationship between the size and age of the individuals, which allowed us to reconstruct population expansion. The overall size structure suggests that the population is still expanding. In the last 50 yr, E. hermaphroditum advanced more than 200 m in the dune system. Expansion started in the 1960s simultaneously at different distances from the shoreline. Colonization did not proceed gradually from the fixed dune toward the shoreline but instead individuals established earlier in the troughs between the dunes, with an increasingly clumped spatial pattern as the population filled in with time. PMID- 21622444 TI - Evolution of floral morphology and pollination system in Bignonieae (Bignoniaceae). AB - The radiation of angiosperms is associated with shifts among pollination modes that are thought to have driven the diversification of floral forms. However, the exact sequence of evolutionary events that led to such great diversity in floral traits is unknown for most plant groups. Here, we characterize the patterns of evolution of individual floral traits and overall floral morphologies in the tribe Bignonieae (Bignoniaceae). We identified 12 discrete traits that are associated with seven floral types previously described for the group and used a penalized likelihood tree of the tribe to reconstruct the ancestral states of those traits at all nodes of the phylogeny of Bignonieae. In addition, evolutionary correlations among traits were conducted using a maximum likelihood approach to test whether the evolution of individual floral traits followed the correlated patterns of evolution expected under the "pollination syndrome" concept. The ancestral Bignonieae flower presented an Anemopaegma-type morphology, which was followed by several parallel shifts in floral morphologies. Those shifts occurred through intermediate stages resulting in mixed floral morphologies as well as directly from the Anemopaegma-type morphology to other floral types. Positive and negative evolutionary correlations among traits fit patterns expected under the pollination syndrome perspective, suggesting that interactions between Bignonieae flowers and pollinators likely played important roles in the diversification of the group as a whole. PMID- 21622445 TI - Legume phylogeny and the evolution of a unique contractile apparatus that regulates phloem transport. AB - Protein bodies called forisomes undergo Ca(2+)-dependent deformations to occlude sieve tubes reversibly, providing a unique regulatory mechanism of phloem transport. Because forisomes are known exclusively from the Papilionoideae (Leguminosae), the evolution of forisome function may have played a role in the rapid radiation of this huge taxon. The unexpected discovery of a papilionoid species lacking forisomes led us to evaluate a representative set of species covering 33 of the 36 legume tribes traditionally recognized. We found forisomes in Papilionoideae but not in Caesalpinioideae and Mimosoideae. Forisomes were absent from several species of the papilionoid tribe Galegeae. Forisomes with tail-like protrusions occurred less frequently than tailless ones; their distribution correlated with taxonomic units but not sharply enough to render forisome type a reliable criterion for classification. Thus, the distribution of forisome types appeared to reflect physiological variability in the pathways of forisome assembly rather than the evolution of forisome genes. On the other hand, Ca(2+)-dependent forisome deformation and sieve tube plugging occurred in Bobgunnia madagascariensis, a member of the swartzioid clade that presumably is the sister group of all other papilionoids, suggesting that forisomes and their unique mechanism of deformation are a synapomorphy of the Papilionoideae. PMID- 21622446 TI - Early Oligocene Callitris and Fitzroya (Cupressaceae) from Tasmania. AB - This paper documents Early Oligocene fossilized foliage and ovulate cones from Lea River, Tasmania and identifies them as belonging to two extant southern hemisphere Cupressaceae genera, Callitris and Fitzroya. Most importantly, it sheds some light on evolutionary trends within Callitris, a genus with numerous extant Australian species and two extant New Caledonian species. Callitris has a very poor fossil record and, because of the present absence of a molecular data set that includes all species, its phylogeny remains somewhat ambiguous. Although Fitzroya foliage has previously been described from a number of Tasmanian sites, this is the first recording of fertile material. The ovulate cones of the Callitris and Fitzroya macrofossils are characterized by bract-scale complexes in two whorls of three and are subtended by scale-like leaves in whorls of three. The fossilized foliage specimens consist of scale-like leaves in whorls of three. These morphological characteristics are only exhibited by three extant southern hemisphere cupressaceous genera, Callitris, Actinostrobus, and Fitzroya. The assignment of the fossils to extinct Callitris and Fitzroya species is made by comparisons with species from these three extant genera. Although much of the Lea River flora are wet rainforest taxa, the Callitris fossils have characteristics of both wet- and dry-adapted extant species. PMID- 21622447 TI - Identity and genetic structure of the photobiont of the epiphytic lichen Ramalina menziesii on three oak species in southern California. AB - Lichens, a classic example of an obligate symbiosis between fungi and photobionts (which could be algae or cyanobacteria), are abundant in many terrestrial ecosystems. The genetic structure of the photobiont population found in association with a lichen-forming fungal species could be affected by fungal reproductive mode and by the spatial extent of gene flow in the photobiont. Using DNA sequences from one nuclear ribosomal and two chloroplast loci, we analyzed the genetic structure of the photobiont associated with the fungus Ramalina menziesii at an oak woodland study site in southern California. We had previously shown that the fungus exhibited no genetic structure among four local sites or three phorophyte species. Our goals were to identify the photobiont species and assess its genetic structure. We found that R. menziesii was highly specific in its photobiont choice and associated with one alga, Trebouxia decolorans. In contrast to the fungal population, we found significant differentiation among the algae sampled on three oak species and little genetic structure among the sites for two of the three algal loci. We hypothesize that R. menziesii is locally adapted to the phorophyte species through habitat specialization in the algal partner of the symbiosis. PMID- 21622448 TI - Population genetic analysis of safflower (Carthamus tinctorius; Asteraceae) reveals a Near Eastern origin and five centers of diversity. AB - Analyses of genetic variation in crop gene pools are a powerful tool for investigating the origin and early evolution of crop lineages. Such analyses also have the potential to identify unique genetic resources for continued crop improvement. The oilseed crop safflower (Carthamus tinctorius) is believed to have been domesticated in the Fertile Crescent region, but up to 10 geographic centers of similarity throughout the world have been proposed based on morphology. Nuclear microsatellite analysis of accessions from each of the 10 proposed centers of similarity, as well as individuals of the progenitor species, suggested the presence of five genetic clusters (1, Europe; 2, Turkey-Iran-Iraq Afghanistan; 3, Israel-Jordan-Syria; 4, Egypt-Ethiopia; and 5, the Far East-India Pakistan). North American accessions, products of a secondary introduction from the native range, suggest that a subset of the native accessions harbor unique genetic diversity that could be useful in future breeding efforts. Overall, a Near Eastern origin of safflower was confirmed based on the genetic similarity between the progenitor and the Near Eastern safflower accessions, as well as previous archaeological finds. Genetic differentiation between geographical clusters of accessions is evident, although not to the degree proposed based on morphology. PMID- 21622449 TI - Cycad mutualist offers more than pollen transport. AB - Specialist insects share obligate mutualisms with some contemporary cycad species whereby the insect's pollination services are rewarded with a nursery in which the insect's larvae consume the postdispersal male cone. I prevented visits of the pollinator moth Anatrachyntis sp. to male Cycas micronesica (Cycadaceae) cones to show that consumption of the cone tissue by the mutualist hastened initiation of the plant's subsequent reproductive event. This is the first documented case where removal of a postdispersal cycad pollination organ speeds up subsequent reproductive events, and the current paradigm that the offering of cone tissue as a nursery is a sacrifice by the plant in return for the pollination services is therefore inaccurate. In C. micronesica, the herbivory stage of pollination mutualism confers a cryptic benefit of cone tissue disposal, which translates into an increase in ultimate lifetime reproductive effort. The plant population relies on the pollinator for moving gametes, as well as for increasing the number of male coning events. The dual benefits afforded to the plant by associating with this pollinator shows that mutualism can operate simultaneously on very different traits. PMID- 21622450 TI - Long-distance pine pollen still germinates after meso-scale dispersal. AB - Viability of long-distance pollen links ecological models to the genetic structure of forest tree populations, determining how forests will adapt to climate change and how far genes flow from genetically modified (GM) pine plantations. Addressing this landscape-scale inquiry is feasible when the pollen source, the delivery system, and the receiver field can be made explicit. To this end, I measured long-distance pollen germination along a 160-km transect along the North Carolina coastline, including 45000 ha of mature Pinus taeda plantations and barrier islands. Using this system, I tested three hypotheses: (1) pine pollen germinates after dispersal on meso-scale distances, (2) sodium chloride exposure reduces germination of pollen captured over open saltwater, and (3) viable pine pollen is present at high altitudes before local peak pollen shed. The experimental findings are as follows: pine pollen had germination rates of 2 to 57% after dispersal at distances from 3 to 41 km, sodium chloride solutions mildly reduced P. taeda pollen germination, and viable pine pollen grains were captured at an altitude of 610 m. GM pine plantings thus have a potential to disperse viable pollen at least 41 km from the source. Wind and rainfall, as integral parts of regional atmospheric systems, together exert a powerful influence on the genetic structure of forest tree populations. PMID- 21622451 TI - Patterns and causes of incongruence between plastid and nuclear Senecioneae (Asteraceae) phylogenies. AB - One of the longstanding questions in phylogenetic systematics is how to address incongruence among phylogenies obtained from multiple markers and how to determine the causes. This study presents a detailed analysis of incongruent patterns between plastid and ITS/ETS phylogenies of Tribe Senecioneae (Asteraceae). This approach revealed widespread and strongly supported incongruence, which complicates conclusions about evolutionary relationships at all taxonomic levels. The patterns of incongruence that were resolved suggest that incomplete lineage sorting (ILS) and/or ancient hybridization are the most likely explanations. These phenomena are, however, extremely difficult to distinguish because they may result in similar phylogenetic patterns. We present a novel approach to evaluate whether ILS can be excluded as an explanation for incongruent patterns. This coalescence-based method uses molecular dating estimates of the duration of the putative ILS events to determine if invoking ILS as an explanation for incongruence would require unrealistically high effective population sizes. For four of the incongruent patterns identified within the Senecioneae, this approach indicates that ILS cannot be invoked to explain the observed incongruence. Alternatively, these patterns are more realistically explained by ancient hybridization events. PMID- 21622452 TI - Migration of endpoints of two genes relative to boundaries between regions of the plastid genome in the grass family (Poaceae). AB - Overlapping genes occur widely in microorganisms and in some plastid genomes, but unique properties are observed when such genes span the boundaries between single copy and repeat regions. The termini of ndhH and ndhF, situated near opposite ends of the small single-copy region (SSC) in the plastid genomes of grasses (Poaceae), have migrated repeatedly into and out of the adjacent inverted-repeat regions (IR). The two genes are transcribed in the same direction, and the 5' terminus of ndhH extends into the IR in some species, while the 3' terminus of ndhF extends into the IR in others. When both genes extend into the IR, portions of the genes overlap and are encoded by the same nucleotide positions. Fine-scale mapping of the SSC-IR junctions across a sample of 92 grasses and outgroups, integrated into a phylogenetic analysis, indicates that the earliest grasses resembled the related taxa Joinvillea (Joinvilleaceae) and Ecdeiocolea (Ecdeiocoleaceae), with ca. 180 nucleotides of ndhH extending into the IR, and with ndhF confined to the SSC. This structure is maintained in early-diverging grass lineages and in most species of the BEP clade. In the PACMAD clade, ndhH lies completely or nearly completely within the SSC, and ca. 20 nucleotides of ndhF extend into the IR. The nucleotide substitution rate has increased in the PACMAD clade in the portion of ndhH that has migrated into the SSC. PMID- 21622453 TI - Bolete diversity in two relict forests of the Mexican beech (Fagus grandifolia var. mexicana; Fagaceae). AB - The current distribution of the endangered Mexican beech [Fagus grandifolia var. mexicana (Martinez) Little] is restricted to relict isolated populations in small remnants of montane cloud forest in northeastern Mexico, and little is known about its associated biota. We sampled bolete diversity in two of these monospecific forests in the state of Hidalgo, Mexico. We compared alpha diversity, including species richness and ensemble structure, and analyzed beta diversity (dissimilarity in species composition) between forests. We found 26 bolete species, five of which are probably new. Species diversity and evenness were similar between forests. Beta diversity was low, and the similarities of bolete samples from within and between forests were not significantly different. These results support the idea that the two forests share a single bolete ensemble with a common history. In contrast, cumulative species richness differed between the forests, implying that factors other than the mere presence of the host species have contributed to shaping the biodiversity of ectomycorrhizal fungi in relict Mexican beech forests. PMID- 21622454 TI - Lower selfing rate at higher altitudes in the alpine plant Eritrichium nanum (Boraginaceae). AB - A general hypothesis on mating patterns in alpine plants states that self fertilization should increase with increasing altitude as a result of pollinator limitation at higher altitudes. However, realized selfing rates under natural conditions, as based on genetic progeny analysis, have not yet been determined for any alpine species across altitude. We therefore assessed the realized selfing rates in about 100 open-pollinated families of the high-alpine cushion plant Eritrichium nanum, sampled along an altitudinal gradient in the Swiss Alps, by using progeny analysis based on six microsatellites. In marked contrast to the general hypothesis, realized selfing rates in E. nanum significantly decreased with increasing altitude, and only progenies from low altitudes were predominantly selfed. However, the higher selfing rates of individuals at lower altitudes could have been caused by unfavorable weather conditions during early growing season when low-elevation plants flowered. In summary, our results on selfing rates in an alpine plant across altitude as well as the results of other studies using experimental hand-pollinations and/or population genetic methods generally do not support the expectation of higher selfing rates at higher altitudes. We therefore ask for further critical examination of realized mating systems in alpine plants. PMID- 21622455 TI - Development of microsatellite loci for Pinus koraiensis (Pinaceae). AB - PREMISE OF THE STUDY: Microsatellite markers were developed for Pinus koraiensis to characterize its genetic diversity and understand its population structure. * METHODS AND RESULTS: Using the Fast Isolation by AFLP of Sequences COntaining (FIASCO) Repeats protocol, 20 primer sets were developed in Chinese populations of P. koraiensis. Three of the markers showed polymorphism with two alleles per locus when assessed in a sample of two populations of P. koraiensis from the Changbai Mountain in the Jilin Province of China. Five and three loci were successfully amplified in P. taiwanensis and P. massoniana, respectively. The amplification size of these loci matches those in P. koraiensis. * CONCLUSIONS: These markers may be useful for further investigation of population genetics of P. koraiensis. PMID- 21622456 TI - Microsatellite markers in the tree peony, Paeonia suffruticosa (Paeoniaceae). AB - PREMISE OF THE STUDY: Microsatellite primers were developed in the tree peony, Paeonia suffruticosa, to perform paternity tests as well as assignment to variety in special Austrian collections. * METHODS AND RESULTS: Using SSR-enriched libraries and EST-mining, 8 polymorphic primer sets were identified in Austrian collections of Paeonia sect. Moutan DC. The primers amplified di- and trinucleotide repeats with 2-6 alleles per locus. All primers also amplified in P. ostii, P. pontaninii var. trolloides, P. delavayi, and P. lutea, and in the herbaceous species P. peregrina and P. tenuifolia (Paeonia sect. Paeon). * CONCLUSIONS: These results show the usefulness of primers in P. suffruticosa for population genetic studies and their ability to cross amplify in related taxa across the genus. PMID- 21622457 TI - Development of a novel set of EST-SSR markers and cross-species amplification in Tamarix africana (Tamaricaceae). AB - PREMISE OF THE STUDY: Tamarix plants are resistant to abiotic stresses and have become invasive in North America. Their taxonomy is troublesome, and few molecular makers are available to enable species identification or to track the spread of specific invasive genotypes. Transcriptome sequencing projects offer a potential source for the development of new markers. * METHODS AND RESULTS: Thirteen polymorphic simple sequence repeats (SSRs) markers derived from Expressed Sequence Tags (ESTs) from Tamarix hispida, T. androssowii, T. ramosissima, and T. albiflonum were identified and screened on 24 samples of T. africana to detect polymorphism. The number of alleles per locus ranged from two to eight, with an average of 4.3 alleles per locus, and the mean expected heterozygosity was 0.453. * CONCLUSIONS: Amplification products of these 13 loci were also generated for T. gallica. These new EST-SSR markers will be useful in genetic characterization of Tamarix, as additional tools for taxonomic clarification, and for studying invasive populations where they are a threat. PMID- 21622458 TI - Development, characterization, and transferability of microsatellite markers for Kirengeshoma palmata (Hydrangeaceae). AB - PREMISE OF THE STUDY: Microsatellite markers were developed for Kirengeshoma palmata to assess the population genetics and mating pattern of this critically endangered species. * METHODS AND RESULTS: A total of 24 microsatellite markers were developed for K. palmata using an enrichment protocol. These markers were screened in 37 individuals from four populations in China and Japan, and twelve were found to be polymorphic, with the number of alleles per locus ranging from two to eight. All of these primers also amplified in K. koreana. * CONCLUSIONS: These microsatellite markers provide a useful tool to investigate the mating system, gene flow, parentage, and population dynamics of Kirengeshoma. PMID- 21622459 TI - Primer development for the plastid region ycf1 in Annonaceae and other magnoliids. AB - PREMISE OF THE STUDY: Primers were developed for a portion of the ycf1 plastid gene in magnoliid taxa to investigate the utility of ycf1 in phylogenetic analyses. * METHODS AND RESULTS: Twenty-six species across six families within the magnoliid group (Canellales, Piperales, Laurales, and Magnoliales) were sampled to examine the ability to amplify ycf1. Additionally, 29 accessions of Asimina and Deeringothamnus (Annonaceae) were sequenced to assess levels of variation in ycf1 compared to matK and trnL-F. * CONCLUSIONS: Results indicate that ycf1 is easily amplified and sequenced. In Annonaceae, ycf1 provides more informative phylogenetic characters than commonly used markers such as matK and trnL-F. PMID- 21622460 TI - Microsatellite primers for Halophila ovalis and cross-amplification in H. minor (Hydrocharitaceae). AB - * PREMISE OF THE STUDY: Polymorphic microsatellite primers were developed in the seagrass Halophila ovalis to investigate genetic variation. * METHODS AND RESULTS: Ten polymorphic microsatellite loci were developed in Halophila ovalis. The number of alleles per locus ranged from 2 to 12 across 80 H. ovalis individuals. These loci were successfully amplified in H. minor, and four were monomorphic across 30 individuals. * CONCLUSIONS: These results from four H. ovalis populations and one H. minor population show the broad utility of microsatellite loci in future studies of population genetics. Four distinct alleles were present in H. minor but absent in H. ovalis, indicating potential divergence between them. PMID- 21622461 TI - C and N stable isotope signatures reveal constraints to nutritional modes in orchids from the Mediterranean and Macaronesia. AB - We compared the nutritional modes and habitats of orchids (e.g., autotrophic, partially or fully mycoheterotrophic) of the Mediterranean region and adjacent islands of Macaronesia. We hypothesized that ecological factors (e.g., relative light availability, surrounding vegetation) determine the nutritional modes of orchids and thus impose restrictions upon orchid distribution. Covering habitats from dark forests to open sites, orchid samples of 35 species from 14 genera were collected from 20 locations in the Mediterranean and Macaronesia to test for mycoheterotrophy. Mycorrhizal fungi were identified via molecular analyses, and stable isotope analyses were applied to test whether organic nutrients are gained from the fungal associates. Our results show that orchids with partial or full mycoheterotrophy among the investigated species are found exclusively in Neottieae thriving in light-limited forests. Neottioid orchids are missing in Macaronesia, possibly because mycoheterotrophy is constrained by the lack of suitable ectomycorrhizal fungi. Furthermore, most adult orchids of open habitats in the Mediterranean and Macaronesia show weak or no N gains from fungi and no C gain through mycoheterotrophy. Instead isotope signatures of some of these species indicate net plant-to-fungus C transfer. PMID- 21622462 TI - Impact of growth form and carbohydrate reserves on tolerance to simulated deer herbivory and subsequent recovery in Liliaceae. AB - PREMISE OF THE STUDY: Over-browsing of the understory vegetation by white-tailed deer has been a cause of decline in many plant populations. Liliaceae are particularly sensitive, yet individual species differ in their tolerance to deer herbivory. In this paper, we examine whether differences in clonal habit, carbon allocation patterns, and phenology influence the capacity of a species to tolerate and recover from repeated herbivory. * METHODS: Flowering ramets of Clintonia borealis, Maianthemum canadense, and Trillium erectum were subjected to total defoliation for one or two springs. * KEY RESULTS: Survival was highest in the nonclonal species, T. erectum, most probably due to its very large carbohydrate reserves. Nutrient reserves were less affected than carbohydrate reserves by defoliation, confirming the importance of carbohydrate reserves for survival. However, faster recovery following episodes of defoliation was observed not in the species that sprouted the earliest, T. erectum, but in the clonal species, M. canadense, which had the smallest carbohydrate reserves but also a lower shoot to root ratio than the other clonal species, C. borealis. All plants that were defoliated for 2 years only partially recovered in terms of leaf area, plant biomass, and carbohydrate and nutrient reserves, confirming the overall sensitivity of these species to simulated deer herbivory. * CONCLUSIONS: High carbohydrate reserves and consequently low shoot to root ratios appear to increase tolerance to herbivory, whereas clonal species recover faster than nonclonal species. The role played by carbohydrates reserves suggests that these species could benefit from slightly higher light conditions in areas subjected to high deer pressure. PMID- 21622463 TI - Photosynthetic and growth responses of reciprocal hybrids to variation in water and nitrogen availability. AB - PREMISE OF THE STUDY: Fitness of plant hybrids often depends upon the environment, but physiological mechanisms underlying the differential responses to habitat are poorly understood. We examined physiological responses of Ipomopsis species and hybrids, including reciprocal F(1)s and F(2)s, to variation in soil moisture and nitrogen. * METHODS: To examine responses to moisture, we subjected plants to a dry-down experiment. Nitrogen was manipulated in three independent experiments, one in the field and two in common environments. * KEY RESULTS: Plants with I. tenuituba cytoplasmic background had lower optimal soil moisture for photosynthesis, appearing better adapted to dry conditions, than plants with I. aggregata cytoplasm. This result supported a prediction from prior studies. The species and hybrids did not differ greatly in physiological responses to nitrogen. An increase in soil nitrogen increased leaf nitrogen, carbon assimilation, integrated water-use efficiency, and growth, but the increases in growth were not mediated primarily by an increase in photosynthesis. In neither the field, nor in common-garden studies, did physiological responses to soil nitrogen differ detectably across plant types, although only I. aggregata and hybrids increased seed production in the field. * CONCLUSIONS: These results demonstrate differences in photosynthetic responses between reciprocal hybrids and suggest that water use is more important than nitrogen in explaining the relative photosynthetic performance of these hybrids compared to their parents. PMID- 21622464 TI - Natural selection on inflorescence color polymorphisms in wild Protea populations: The role of pollinators, seed predators, and intertrait correlations. AB - PREMISE OF THE STUDY: The processes maintaining flower color polymorphisms have long been of evolutionary interest. Mechanistic explanations include selection through pollinators, antagonists, local environments, drift, and pleiotropic effects. We examined the maintenance of inflorescence color polymorphisms in the genus Protea (Proteaceae) of South Africa, in which ~40% of species contain different color morphs. * METHODS: We studied 10 populations of four bird pollinated Protea species and compared adult performance, floral and leaf morphology, vegetative pigmentation, germination, and seedling survival between co-occurring pink and white morphs. We also tested for differences in pollination success and pre-dispersal seed predation. * KEY RESULTS: White morphs produced seeds 10% heavier and 3.5 times more likely to germinate, which all else being equal, should fuel positive selection on white. In one studied population per species, however, white morphs were more susceptible to seed predation by endophagous larvae. Pollinators had no morph-specific effects on female fecundity, as measured by amount or probability of seed set. Differences in stem color indicated that white morphs produced smaller quantities of pigment and associated compounds throughout, possibly explaining their higher seed palatability. * CONCLUSIONS: Our findings suggest a mechanism for some white protea polymorphisms: deleterious pleiotropic effects on pink morphs are occasionally offset by reduced losses to seed-eating larvae. Because trends were repeated across species, we suggest that similar processes may also occur in other proteas, placing a new emphasis on seed predators for influencing some of South Africa's amazing floral diversity. PMID- 21622465 TI - Hybridization and introgression in New World red mangroves, Rhizophora (Rhizophoraceae). AB - PREMISE OF THE STUDY: Hybridization is common in both animals and plants and can lead to a diverse array of outcomes ranging from the generation of new ecotypes or species to the breakdown of morphological differences. Here, we explore the extent of hybridization in the three currently recognized New World Rhizophora species-R. mangle, R. racemosa, and the putative hybrid species R. harrisonii. * METHODS: We assayed variation across the three recognized Rhizophora species using two noncoding chloroplast (cpDNA), two flanking microsatellite regions (FMRs), and six microsatellite loci. * KEY RESULTS: Gene genealogies of cpDNA and FMRs showed a strong phylogeographic break across the Central American Isthmus, but little relationship to recognized species boundaries. Instead, individuals collected in the same ocean basin and classified as R. mangle and R. racemosa by morphological characteristics were more closely related to each other than with similar looking individuals collected in the other ocean basin. Nonetheless, there were low, yet significant differences at microsatellite loci among co occurring populations of R. mangle and R. racemosa in both ocean basins, suggesting that two taxonomic groups coexist. However, we found no genetic evidence that R. harrisonii was a hybrid species. Rather, R. harrisonii appears to represent a morphotype produced by ongoing hybridization and backcrossing between R. mangle and R. racemosa. * CONCLUSIONS: Our data support ancient and persistent introgressive hybridization among new world Rhizophora and argue for a full revision of the systematic relationships of the group based on much finer morphological, ecological, and genetic analyses. PMID- 21622466 TI - Speciation pathway of Isoetes (Isoetaceae) in East Asia inferred from molecular phylogenetic relationships. AB - Polyploidy plays an important role in the speciation of Isoetes. Increasing our knowledge about the specific origin of each polyploid or phylogenetic relationship among species has been hampered because of conserved morphological variation and scarce habitats. We present several hypotheses concerning the speciation pathways of Isoetes species distributed in East Asia. Our hypotheses are inferred from phylogenetic relationships that were elucidated using sequences of the internal transcribed spacer regions of nuclear ribosomal DNA, a second intron of LEAFY, and chloroplast DNA trnS-psbC spacer regions. These inferred phylogenetic relationships indicated that (1) the Chinese tetraploid, I. sinensis, is closely related to I. yunguiensis; (2) the Korean endemic species, I. hallasanensis, is an autotetraploid derived from I. taiwanensis or closely related taxa; (3) the hexaploid I. coreana forms a clade and has its closest evolutionary relationships with I. taiwanensis or I. hallasanensis; and (4) the Japanese hexaploid I. japonica is closely related to I. taiwanensis-I. coreana and I. sinensis-I. yunguiensis. These results suggest that interspecific hybridization and polyploidization have played central roles in speciation of East Asian Isoetes. Furthermore, I. taiwanensis, an endemic species in Taiwan, has been involved in at least three cases of autopolyploid or allopolyploid speciation in East Asia. PMID- 21622467 TI - Projections of suitable habitat for rare species under global warming scenarios. AB - PREMISE OF THE STUDY: Modeling the contemporary and future climate niche for rare plants is a major hurdle in conservation, yet such projections are necessary to prevent extinctions that may result from climate change. * METHODS: We used recently developed spline climatic models and modified Random Forests statistical procedures to predict suitable habitats of three rare, endangered spruces of Mexico and a spruce of the southwestern USA. We used three general circulation models and two sets of carbon emission scenarios (optimistic and pessimistic) for future climates. * KEY RESULTS: Our procedures predicted present occurrence perfectly. For the decades 2030, 2060, and 2090, the ranges of all taxa progressively decreased, to the point of transient disappearance for one species in the decade 2060 but reappearance in 2090. Contrary to intuition, habitat did not develop to the north for any of the Mexican taxa; rather, climate niches for two taxa re-materialized several hundred kilometers southward in the Trans Mexican Volcanic Belt. The climate niche for a third Mexican taxon shrank drastically, and its two mitotypes responded differently, one of the first demonstrations of the importance of intraspecific genetic variation in climate niches. The climate niche of the U.S. species shrank northward and upward in elevation. * CONCLUSION: The results are important for conservation of these species and are of general significance for conservation by assisted colonization. We conclude that our procedures for producing models and projecting the climate niches of Mexican spruces provide a way for handling other rare plants, which constitute the great bulk of the world's endangered and most vulnerable flora. PMID- 21622468 TI - Microsatellite markers show decreasing diversity but unchanged level of clonality in Dryas octopetala (Rosaceae) with increasing latitude. AB - PREMISE OF THE STUDY: Average arctic temperatures have increased at almost twice the global average in the past 100 years. Most studies on biodiversity along latitudinal gradients have focused on species richness or genetic diversity at lower latitudes, and only a few studies have inferred genetic diversity within a species along a latitudinal gradient at higher latitudes, even though these areas might be most affected by recent climate changes. Here, intraspecific genetic diversity of the arctic-alpine Dryas octopetala (Rosaceae) is studied along a latitudinal gradient to test the hypotheses that genetic diversity decreases and vegetative clonal growth increases with latitude. * METHODS: Ten microsatellite markers have been developed for D. octopetala and analyzed with population genetic methods in five populations along a latitudinal transect spanning from 59.0 degrees N to 79.9 degrees N. * KEY RESULTS: The nine microsatellites that were used in the final analyses resulted in a resolution high enough to distinguish between ramets while providing useful information at a larger geographical scale. Three genetic clusters were indicated, a southern Norway group, a northern Norway group, and a Svalbard group, with corresponding decreasing genetic diversity. No trend was found with regard to clonality along the gradient. * CONCLUSIONS: The newly developed microsatellite markers provide a useful tool for further genetic studies of D. octopetala and its close relatives, addressing population structure as well as phylogeographic patterns. The results of this study support the hypothesis of decreasing genetic diversity with increasing latitude, which may have implications for future adaptability to climate change. PMID- 21622469 TI - Reconstructing Dryopteris "semicristata" (Dryopteridaceae): Molecular profiles of tetraploids verify their undiscovered diploid ancestor. AB - PREMISE OF THE STUDY: Discovering missing ancestors is essential to understanding the evolutionary history of biodiversity on Earth. Evidence from extinct species can provide links for reconstructing intricate patterns of reticulate relationships among extant descendents. When fossils are unavailable and other evidence yields competing hypotheses to explain species ancestry, data from proteins and DNA can help resolve conflicts and generate novel perspectives. The identity of a parent shared by two tetraploid species in the cosmopolitan fern genus Dryopteris has remained elusive for more than 50 years. Based on available data, four hypotheses were developed previously, each providing a different resolution to this uncertainty. * METHODS: New molecular evidence from studies of isozymes and restriction site analysis of chloroplast DNA tested the competing hypotheses about the diploid ancestors of these two extant Dryopteris polyploids. * KEY RESULTS: The results falsify two of the hypotheses, resolve the uncertainty in the third, and support the fourth. * CONCLUSIONS: Our data validate the prior existence of Dryopteris "semicristata," which was proposed 38 years ago as a diploid progenitor of the allotetraploids D. cristata and D. carthusiana but has never been collected. After developing a phylogeny using the new molecular data, we describe a plausible morphology for D. "semicristata" by extrapolating likely character states from related extant species. PMID- 21622470 TI - Mixed mating in a recently derived self-compatible population of Leavenworthia alabamica (Brassicaceae). AB - PREMISE OF THE STUDY: A mixture of outcrossing and selfing is often observed in plant populations. Although mixed mating is ubiquitous, it has several potential evolutionary explanations. Mixed mating may be actively maintained by selection, passively determined by the pollination environment, or a transitional stage during the evolution of self-fertilization. * METHODS: We studied patterns of self-compatibility and selfing rates in a population of Leavenworthia alabamica that recently lost self-incompatibility. We also experimentally tested whether natural selection against selfing at the pre- or postzygotic stage is sufficient to explain mixed mating in this population. * KEY RESULTS: Visualizing pollen tube growth following self-pollination, we found that nearly all plants were fully self-compatible. Progeny array analysis revealed that the average selfing rate of the population was s = 0.523. The inbreeding coefficient in the parents (F = 0.539) exceeded the amount expected if the selfing rate (s) were constant [F(eq) = s/(2 - s)], indicating either population subdivision or higher selfing rates in the past. Inference of family-level selfing rates revealed substantial variation. Experiments found that self and outcross pollen fertilized nearly equal numbers of ovules in competition. Comparison of seed production following self- or cross-pollination failed to implicate early acting inbreeding depression as a factor maintaining mixed mating. * CONCLUSIONS: The results of our experiments suggest that mixed mating is not maintained by selection against self pollen or zygotes in this population. Mixed mating is most likely a byproduct of the pollination process but may also be a transitional stage during the evolution of higher selfing rates. PMID- 21622471 TI - Fine-scale comparisons of genetic variability in seed families of asexually and sexually reproducing Crataegus (hawthorn; Rosaceae). AB - The reproductive system is one of the key mechanisms that determine genetic diversity at different biological levels. However, few attempts have been made to assess the consequences of asexual reproduction by comparing genetic structure and fecundity of seed families in asexually and sexually reproducing individuals. We have examined two similar hawthorn species, Crataegus crus-galli and C. punctata, that differ in ploidy level and breeding system. Fecundity (per-fruit seed set) and microsatellite genotypes for five loci were determined in 18 and 26 seed families of C. crus-galli and C. punctata (totals of 83 and 118 embryos), respectively. Compared with the sexual diploid C. punctata, tetraploid C. crus galli shows similar fecundity, but lower genotypic diversity within and between seed families. Reproduction in the tetraploid, while predominantly apomictic, is nevertheless accompanied by outcrossing and selfing. We conclude that in C. crus galli pollen flow between conspecific individuals is limited, and the combination of pollen fertility, self-compatibility, and pseudogamous apomixis provides reproductive assurance in these tetraploids. Reproductive assurance, in turn, may explain the derived floral architecture seen in most North American tetraploid hawthorns. We also discuss analytical approaches for inferring mating-system parameters in tetraploids and for comparing microsatellite variation across ploidy levels. PMID- 21622472 TI - Antimicrobial nectar inhibits a florally transmitted pathogen of a wild Cucurbita pepo (Cucurbitaceae). AB - PREMISE OF THE STUDY: Floral nectars of many species contain antimicrobial chemicals, but their function in nectar is subject to debate. Previously, we have shown that Erwinia tracheiphila, the causative agent of bacterial wilt disease in cucurbits, can be transmitted via the floral nectaries. * METHODS: We used a disk diffusion assay (DDA) to determine the antimicrobial effects of nectar from a wild gourd on lawns of Escherichia coli and Erwinia tracheiphila. We also used E. tracheiphila to inoculate flowers of wild gourd plants, with and without nectar. * KEY RESULTS: The DDA showed that paper disks saturated with 10 MUL of nectar inhibited the growth of E. coli on a larger area of the lawn than 40% glucose but a smaller area than 5% ampicillin for 12 h. On lawns of E. tracheiphila, nectar inhibited growth on a larger area than glucose for 24 h and there were no significant differences between ampicillin and nectar for12 h. A significantly larger proportion of the plants inoculated via flowers without nectar contracted wilt disease than plants with nectar. * CONCLUSIONS: These findings indicate that nectar reduces transmission of E. tracheiphila via the nectaries and reveal the potential for florally transmitted pathogens to influence the evolution of floral traits. PMID- 21622473 TI - Breeding system variation in 10 evening primroses (Oenothera sections Anogra and Kleinia; Onagraceae). AB - PREMISE OF THE STUDY: We examined two accounts of the relationship between breeding system and life history variation in a clade of evening primroses (Oenothera, Onagraceae): (1) selection for reproductive assurance should generate an association between self-compatibility and monocarpy and (2) phylogenetic conservatism leads to retention of breeding system and life history traits among closely related taxa. * METHODS: We performed over 4000 hand pollinations under greenhouse conditions to determine the compatibility of 10 Oenothera taxa (sections Anogra [17 taxa] and Kleinia [2 taxa)] for which breeding systems had not previously been reported. We used generalized linear mixed models to evaluate the influence of pollination treatment, parents, and population on fruiting success. * KEY RESULTS: Among the taxa tested, six were self-incompatible, two were variable in compatibility, and two were self-compatible. We combined these data with published studies in Anogra and Kleinia and mapped breeding system and life history onto a published phylogeny. * CONCLUSIONS: We found no evidence for phylogenetic conservatism, but detected considerable evolutionary lability in both traits. Additionally, we found no evidence for a consistent relationship between breeding system and life history. Only eight of 19 taxa followed the predicted association between self-incompatibility and polycarpy vs. self compatibility and monocarpy. Instead, many taxa have retained self incompatibility, regardless of monocarpy or polycarpy. PMID- 21622474 TI - Colonizing the American continent: Systematics of the genus Arabis in North America (Brassicaceae). AB - The circumscription of the genus Arabis underwent many and drastic changes within the past. Using DNA sequence information from the nuclear ribosomal RNA and parts of the plastid genome (trnL-trnLF), as well as a critical evaluation of herbarium material from East Asia and North America, we circumscribe the various Arabis taxa of North America. The American and East Asian Arabis species are closely related and, contrary to what was previously believed, they are not closely related to the Eurasian A. hirsuta. Using cpDNA, we found five North American lineages of Arabis with distinct distribution patterns, of which only the purple/red-flowered lineage consists of proven diploids that evolved directly from East Asian progenitors. All other four lineages evolved via ancient hybridization either on the Asian continent prior to migration to North America or showed significant evidence for hybridization and reticulation while diversifying on the American continent. We also provide the first evidence for the systematic circumscription of East Asian Arabis taxa, which together with the North American taxa, form one clade distantly related to European A. ciliata and Eurasian A. hirsuta. The findings also represent the first record of A. pycnocarpa for the floras of China, Japan, and Russian Far East. PMID- 21622475 TI - Genetic structure and phylogeography of a temperate-boreal herb, Cardamine scutata (Brassicaceae), in northeast Asia inferred from AFLPs and cpDNA haplotypes. AB - PREMISE OF THE STUDY: Studies on genetic structure of plant populations help us understand the history of local flora and vegetation. In this study, we focus on the temperate-boreal herb Cardamine scutata from northeast Asia, an area with scarce phylogeographic studies. We explore patterns of genetic variation within this species, with an aim to infer its (post-) glacial history with reference to colonization routes and migrations via land bridges. * METHODS: We analyzed 46 populations sampled in Japan, Kamchatka, and Korea using AFLP and cpDNA sequence data. * KEY RESULTS: Two intraspecific genetic groups were resolved, distributed in the northeastern and southwestern part of the study area, most likely reflecting lineages isolated from each other during (at least) the last glaciation. A zone of secondary contacts was found in central/northern Honshu, and a few cases of long-distance dispersal were observed. We detected efficient gene flow across the marine straits, supporting the role of land bridges created by sea level decline during the last glacial period. The cpDNA data indicated extensive recent expansion and diversification within both lineages. We inferred recent colonization of Kamchatka from Hokkaido, associated with genetic impoverishment. * CONCLUSIONS: The pattern of north-south genetic differentiation found in C. scutata is rather common among several other plant species studied in Japan, despite their distinct biological features. We assume that different processes and factors may have brought about this similarity. Overall, this study contributes to better understanding of the biogeography of northeast Asia. PMID- 21622476 TI - Race-ethnicity as an effect modifier of the association between HbAlc and mortality in U.S. adults without diagnosed diabetes. AB - OBJECTIVE: HbAlc is increasingly appreciated as a risk factor for all-cause and cardiovascular disease (CVD) mortality in the non-diabetic population. In this study, we investigated the association between HbAlc and mortality with a particular focus on the impact of race-ethnicity. Design Cohort study. METHODS: We analyzed the association between HbAlc and all-cause and CVD mortality in 12 698 non-diabetic adults 20 years or older from the Third National Health and Nutrition Examination Survey using separate models for people of different race ethnicity. RESULTS: In our stratified analyses, higher non-diabetic HbAlc levels were associated with all-cause and CVD mortality in non-Hispanic whites only. In this group, compared with HbAlc values of 5.0-<5.35%, the multivariable-adjusted estimated hazard ratios (est. HR) with 95% confidence interval (CI) for all-cause mortality were 1.21 (0.92, 1.58), 1.22 (1.03, 1.45), 1.29 (1.14, 1.47), and 1.4 (1.02, 1.87) for HbAlc levels of <5.0, 5.35-<5.7, 5.7-<6.5, and 6.5% or greater respectively. The association did not reach significance in Mexican-Americans (est. HR (95% CI): 1.77 (1.08, 2.91), 0.81 (0.56, 1.19), 1.16 (0.86, 1.57), and 1.4 (0.83, 2.36)). No association was observed in non-Hispanic blacks: 1.13 (0.91, 1.39), 0.81 (0.61, 1.08), 0.84 (0.69, 1.03), and 0.94 (0.67, 1.33). Results were similar for CVD mortality. CONCLUSIONS: Our data suggest limitations of HbAlc as a risk factor for all-cause and cardiovascular mortality across race ethnic populations. PMID- 21622477 TI - A combination of polymorphisms in HSD11B1 associates with in vivo 11{beta}-HSD1 activity and metabolic syndrome in women with and without polycystic ovary syndrome. AB - OBJECTIVE: Regeneration of cortisol by 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) within liver and adipose tissue may be of pathophysiological importance in obesity and the metabolic syndrome. single nucleotide polymorphisms (SNPs) in HSD11B1, the gene encoding 11beta-HSD1, have been associated with type 2 diabetes and hypertension in population-based cohort studies, and with hyperandrogenism in patients with the polycystic ovary syndrome (PCOS). However, the functional consequences of these SNPs for in vivo 11beta-HSD1 expression and activity are unknown. METHODS: We explored associations of well-characterised hormonal and metabolic phenotypes with two common SNPs (rs846910 and rs12086634) in HSD11B1 in 600 women (300 with PCOS) and investigated 11beta-HSD1 expression and activity in a nested study of 40 women from this cohort. RESULTS: HSD11B1 genotypes (as single SNPs and as the combination of the two minor allele SNPs) were not associated with PCOS. Women who were heterozygous for rs846910 A and homozygous for rs12086634 T (GA, TT genotype) had a higher risk of metabolic syndrome, regardless of the diagnosis of PCOS (odds ratio in the whole cohort=2.77 (95% confidence interval (CI) 1.16-6.67), P=0.023). In the nested cohort, women with the GA, TT genotype had higher HSD11B1 mRNA levels in adipose tissue, and higher rates of appearance of cortisol and d3-cortisol (16.1+/-0.7 nmol/min versus 12.1+/-1.1, P=0.044) during 9,11,12,12-2H4-cortisol (d4-cortisol) steady-state infusion. CONCLUSIONS: We conclude that, in a population of Southern European Caucasian women with and without PCOS, alleles of HSD11B1 containing the two SNPs rs846910 A and rs12086634 T confer increased 11beta-HSD1 expression and activity, which associates with the metabolic syndrome. PMID- 21622478 TI - Premature adrenarche: novel lessons from early onset androgen excess. AB - Adrenarche reflects the maturation of the adrenal zona reticularis resulting in increased secretion of the adrenal androgen precursor DHEA and its sulphate ester DHEAS. Premature adrenarche (PA) is defined by increased levels of DHEA and DHEAS before the age of 8 years in girls and 9 years in boys and the concurrent presence of signs of androgen action including adult-type body odour, oily skin and hair and pubic hair growth. PA is distinct from precocious puberty, which manifests with the development of secondary sexual characteristics including testicular growth and breast development. Idiopathic PA (IPA) has long been considered an extreme of normal variation, but emerging evidence links IPA to an increased risk of developing the metabolic syndrome (MS) and thus ultimately cardiovascular morbidity. Areas of controversy include the question whether IPA in girls is associated with a higher rate of progression to the polycystic ovary syndrome (PCOS) and whether low birth weight increases the risk of developing IPA. The recent discoveries of two novel monogenic causes of early onset androgen excess, apparent cortisone reductase deficiency and apparent DHEA sulphotransferase deficiency, support the notion that PA may represent a forerunner condition for PCOS. Future research including carefully designed longitudinal studies is required to address the apparent link between early onset androgen excess and the development of insulin resistance and the MS. PMID- 21622479 TI - fMRI evidence that precision ophthalmic tints reduce cortical hyperactivation in migraine. AB - BACKGROUND: Certain patterns can induce perceptual illusions/distortions and visual discomfort in most people, headaches in patients with migraine, and seizures in patients with photosensitive epilepsy. Visual stimuli are common triggers for migraine attacks, possibly because of a hyperexcitability of the visual cortex shown in patients with migraine. Precision ophthalmic tints (POTs) are claimed to reduce perceptual distortions and visual discomfort and to prevent migraine headaches in some patients. We report an fMRI visual cortical activation study designed to investigate neurological mechanisms for the beneficial effects of POTs in migraine. METHODS: Eleven migraineurs and 11 age- and sex-matched non headache controls participated in the study using non-stressful and stressful striped patterns viewed through gray, POT, and control coloured lenses. RESULTS: For all lenses, controls and migraineurs did not differ in their response to the non-stressful patterns. When the migraineurs wore gray lenses or control coloured lenses, the stressful pattern resulted in activation that was greater than in the controls. There was also an absence of the characteristic low-pass spatial frequency (SF) tuning in extrastriate visual areas. When POTs were worn, however, both cortical activation and SF tuning were normalized. Both when observing the stressful pattern and under more typical viewing conditions, the POTs reduced visual discomfort more than either of the other two lenses. CONCLUSION: The normalization of cortical activation and SF tuning in the migraineurs by POTs suggests a neurological basis for the therapeutic effect of these lenses in reducing visual cortical hyperactivation in migraine. PMID- 21622480 TI - A halo may lead the way. PMID- 21622481 TI - Changes in active life expectancy among older thais: results from the 1997 and 2004 national health examination surveys. AB - This study aims to determine the disability prevalence and changes in active life expectancy of the Thai older people between 1997 and 2004. Data on disability of older people aged >=60 years were obtained from the National Health Examination Surveys. Disability refers to one or more restrictions on the activities of daily living. The Sullivan method was used to calculate active life expectancy. A total of 4048 older people in 1997 and 19 372 older people in 2004 were included in the analysis. Active life expectancy at age 60 of men was 16.5 years in 1997 and 17.6 years in 2004, whereas that of women was 17.9 and 19.9 years, respectively. Women spent a greater proportion of the remaining life with disability. The proportion of active life for both genders also increased during the 7-year period suggesting an evidence of the compression of morbidity in Thai older people. PMID- 21622482 TI - Assessing the five factors of personality in adolescents: the junior version of the Spanish NEO-PI-R. AB - This article presents the development of a junior version of the Spanish (Castilian) NEO Personality Inventory-Revised (JS NEO) suitable for adolescents aged 12 to 18 years. The psychometric properties of the new JS NEO were investigated using two samples of 2,733 and 983 adolescents in Spain. The results showed that the adult NEO-PI-R factor structure was replicated with the junior version of the inventory and that the reliabilities of the scales were adequate. The cross-form correlations between the junior and the adult versions of the questionnaires indicated good equivalence indices. Furthermore, a joint factor analysis of the JS NEO and the Big Five Questionnaire-Children (BFQ-C) provided additional evidence for the construct validity of the JS NEO. PMID- 21622483 TI - Effects of low-level arsenic exposure on urinary N-acetyl-beta-D-glucosaminidase activity. AB - This study was aimed to evaluate whether renal tubular function is impaired by exposure to relatively low concentrations of arsenic. Mean urinary arsenic concentrations and N-acetyl-beta-D-glucosaminidase (NAG) activities were compared among 365 and 502 Korean men and women, respectively, in relation to gender, smoking, alcohol consumption, and recent seafood consumption. The study subjects were divided into 4 groups according to urinary NAG activity and seafood consumption prior to urine sampling, and the correlation between arsenic concentration and urinary NAG activity was tested for each group. The mean urinary arsenic level was higher in women, non-smokers, and non-drinkers in comparison to men, smokers, and drinkers, respectively. Individuals who consumed seafood within 3 days prior to urine sampling showed a higher mean urinary arsenic level than those who did not. The correlation between urinary arsenic concentration and NAG activity in urine was significant only in subjects who did not consume seafood within 3 days prior to urine sampling and whose urinary NAG activity was 7.44 U/g creatinine (75th percentile) or higher. The urinary arsenic concentration was a significant determinant of urinary NAG activity in subjects with NAG activity higher than 7.44 U/g creatinine and especially in those who had not consumed seafood recently. These facts suggest that a relatively low-level exposure to inorganic arsenic produces renal tubular damage in humans. PMID- 21622484 TI - Severe pretibial myxedema. AB - Although pretibial myxedema (PTM) occurs in 4.3% of patients with Graves's disease, the most severe variant, elephantiasis nostras verrucosa, is found in less than 1% of cases. The most frequent location of infiltration is the lower extremities, especially the pretibial areas and on the dorsum of the foot. The authors report one of the most severe cases of elephantiasis nostras verrucosa, following radioactive iodine therapy. PMID- 21622485 TI - Seminar review: sociocultural practices and epidemiology of diabetic foot problem: lessons from a study in Chiang Mai University Hospital, Thailand. AB - The data on sociocultural practices and epidemiology of diabetic foot problems are scarce in Thailand. This report used data found in a database with patients involved in multidisciplinary foot care. Of the 511 patients with diabetes, 475 (93.0%) patients had type 2 diabetes. The prevalence of foot ulcers in diabetic patients was 12.5% and the amputation rate was 1.4%. A total of 32.7% of patients suffered from neuropathic problems. Barefoot walking inside the house was found 55.4% of the time, and this seemed to link closely with Thai sociocultural practices. Improvement in foot care was a direct result of foot care education. PMID- 21622486 TI - Opioid and adjuvant analgesics: compared and contrasted. AB - An adjuvant (or co-analgesic) is a drug that in its pharmacological characteristic is not necessarily primarily identified as an analgesic in nature but that has been found in clinical practice to have either an independent analgesic effect or additive analgesic properties when used with opioids. The therapeutic role of adjuvant analgesics (AAs) is to increase the therapeutic index of opioids by a dose-sparing effect, add a unique analgesic action in opioid-resistant pain, or reduce opioid side effects. A notable difference between opioids and AAs is that unlike opioids some AAs are associated with permanent organ toxicity, for example, nonsteroidal anti-inflammatory drugs (NSAIDs) and renal failure. It is impossible to predict in advance in a given individual what opioid dose they may require to control cancer pain. Most AAs have a ceiling effect for their analgesic actions, but often with continued dose related toxicities and side effects (with the exception of glucocorticoids). The blood levels of opioids (and their metabolites) can be measured with great precision and accuracy. There is sometimes a role for drug blood levels of certain AAs, like tricyclic antidepressants or anticonvulsants when used for neuropathic pain. Age affects metabolism of most opioids. The therapeutic window of opioids is wide, with no ceiling effect. Most AAs (except corticosteroids) have a narrow therapeutic window. Naloxone is a pure opioid antagonist that competes and displaces opioids from their receptor sites. All clinically useful opioids are mu opioid receptor agonists. Not all routes of administration are available to all opioids. Adjuvant analgesics lack the versatility in routes of administration that opioids possess. Dosing flexibility is a major advantage when treating cancer-related pain with opioids. Dose flexibility is much less with AAs than opioids. Unlike opioids, the analgesic response is usually observed within hours to days of attaining an adequate dose with most AAs (1-2 days). Rotation among opioids is a useful therapeutic strategy to improve analgesic response or minimize toxicity. Most AAs are unsuitable for rescue dosing because of their pharmacological characteristics. The mu agonist side effect profile is similar among the different opioid agents, regardless of the route of administration. The appropriate use of AAs will reduce opioid-related side effects. No apparent tolerance to analgesia develops with AAs. Abrupt discontinuation of an opioid after chronic repeated use for more than a few days will cause a withdrawal syndrome of variable severity. Adjuvant analgesics are an essential tool in cancer pain. PMID- 21622488 TI - Oxycodone and a useful book. PMID- 21622487 TI - Synaptocrine signaling: steroid synthesis and action at the synapse. AB - Sex steroids have long been recognized for their dramatic impact on brain and behavior, including rapid modulation of membrane excitability. It is a widely held perception that these molecules are largely derived from peripheral sources and lack the spatial and temporal specificity ascribed to classical neuromodulatory systems. Neuromodulatory systems, in contrast, are defined by their regulated neuronal presynaptic secretion and by their functional modulation of perisynaptic events. Here we provide evidence for regulated presynaptic estrogen synthesis and functional postsynaptic actions. These results meet all the criteria for a neuromodulatory system and shift our perception of estrogens from that of peripheral signals exclusively to include that of a signaling system intrinsic to the brain itself. We apply the term synaptocrine to describe this form of neuromodulation. PMID- 21622489 TI - US doctors identify top five activities that can be reduced. PMID- 21622490 TI - Three out of 12 hospitals fail to meet essential standards of care of older people, finds watchdog. PMID- 21622491 TI - Socioeconomic patterning in tobacco use in Argentina, 2005. AB - INTRODUCTION: Globally, tobacco is the number one preventable cause of death, killing 1 in 10 adults. By 2030, 80% of all tobacco deaths will occur in developing countries. Social factors drive the adoption and cessation of smoking in high-income countries, but few studies have examined the socioeconomic patterning of smoking in developing countries. METHODS: Using data from the 2005 National Survey of Risk Factors for Non-communicable Diseases in Argentina, we investigated gender-specific socioeconomic patterning of current, former, and never-smoking status; the intensity of smoking; and smokers' readiness for cessation using multinomial logistic and ordinary least squares regression. We also investigated heterogeneity in the patterning by age. RESULTS: Higher socioeconomic position (SEP) was associated with less smoking for men in all age groups, although the results were most pronounced for men at younger ages (odds ratio [OR] of current vs. never smoking = 0.57, 95% CI 0.51-0.63 for higher vs. lower education at ages 18-24 years). For women, higher SEP was associated with more smoking in older age groups but less smoking in younger age groups (OR = 1.51, CI 1.41-1.62 and OR = 0.86, CI 0.78-0.96 for higher vs. lower education at ages 50-64 and 18-24 years, respectively). Higher SEP was also associated with higher odds of recently quitting compared to not considering quitting for men regardless of age group but for women only in younger age groups. DISCUSSION: The higher burden of smoking among those of lower SEP, especially in younger age groups, is a troubling pattern that is certain to impact future disparities in chronic disease outcomes unless interventions are undertaken. PMID- 21622492 TI - An investigation of social and pharmacological exposure to secondhand tobacco smoke as possible predictors of perceived nicotine dependence, smoking susceptibility, and smoking expectancies among never-smoking youth. AB - INTRODUCTION: Recent studies evidenced that adolescent never-smokers exposed to secondhand tobacco smoke (SHS) endorsed nicotine dependence symptoms. Other studies showed that SHS exposure measured with biomarkers among never-smokers independently predicted withdrawal sensations and prospective smoking initiation. The aim of the present study was to replicate and extend these findings by investigating whether social and pharmacological measures of SHS exposure predicted precursors to smoking among never-smoking adolescents. METHODS: Participants included 327 never-smokers aged 11-15 years attending sixth or seventh grade in French language schools in Montreal, Canada. They completed self report questionnaires measuring their smoking status, social smoke exposure (number of smokers in their environment and number of situations where SHS exposure occurs), and precursors to smoking initiation (smoking expectancies, perceived nicotine dependence, and smoking susceptibility). Each participant provided a saliva sample from which cotinine biomarkers were derived to measure pharmacological exposure to SHS. RESULTS: When predictors were modeled individually, number of smokers predicted perceived nicotine dependence (p <= .05), smoking susceptibility (p <= .001), and expected benefits (p <= .05), whereas number of situations predicted smoking susceptibility (p <= .01). When predictors were modeled simultaneously, number of smokers predicted perceived nicotine dependence (p <= .01), smoking susceptibility (p <= .01), and expected benefits (p <= .05). CONCLUSIONS: Social smoke exposure was a predictor for smoking precursors. Pharmacological exposure to SHS did not predict smoking precursors, which may be partly attributable to the low cotinine values observed in our sample. Suggestions for improved pharmacological measurement of SHS and implications for public health are discussed. PMID- 21622493 TI - Smoking and predictors of nicotine dependence in a homeless population. AB - OBJECTIVE: To assess prevalence rates of tobacco use and dependence in a sample of homeless individuals and to investigate trends for demographic and clinical characteristics across different levels of nicotine dependence (nonsmokers vs. lowly dependent smokers vs. highly dependent smokers). METHODS: A cross-sectional study of 489 homeless men and women in 3 Canadian cities. Each subject was assessed using structured clinical interviews and the Fagerstrom Test for Nicotine Dependence (FTND). Cochran-Armitage trend tests were applied to determine unadjusted trends in sociodemographic and clinical variables across levels of nicotine dependence. A generalized logit model was computed to adjust for potential confounding. RESULTS: The mean age was 37.9 years; 39.2% of the participants were women. About 80.8% were current smokers; the mean FTND score was 5.0. Although no significant differences were found between nonsmokers and smokers with low nicotine dependence, smokers with high nicotine dependence were only half as likely as nonsmokers to be Aboriginal, were 2.39 times more likely to have ever been incarcerated, and 2.44 times more likely to have current drug dependence. There were significant trends for the use of cocaine, opioids, and alcohol, with nonsmokers having the lowest and highly dependent smokers having the highest rates of using these substances. CONCLUSIONS: Available public health smoking cessation treatment opportunities should be made available within health care services for the homeless. There is also a need for developing and implementing tobacco dependence treatment programs, which are accessible and tailored to meet the needs of this specific population, accounting for polysubstance use and concurrent substance dependence and mental health disorders. PMID- 21622494 TI - Duration of smoking abstinence and suicide-related outcomes. AB - OBJECTIVE: To investigate the association between suicide-related outcomes (SROs: wish to die, suicidal thoughts, and attempted suicide) and duration of smoking abstinence. METHODS: The National Epidemiologic Survey on Alcohol and Related Conditions Wave 1 is a face-to-face survey of a representative sample of the U.S. adult population (N = 43,093). Analyses were done for a subsample of individuals (N = 13,691) who reported ever smoking, at least 2 weeks of lifetime depressed mood and SROs. Duration of abstinence was categorized as 1-24 hr (reference), 1 day to 12 months, and longer than 12 months. RESULTS: Univariate analyses showed significant demographic associations (positive: female gender and being widowed/divorced/separated; negative: age and household income) with SROs. SROs were positively associated with major psychiatric disorders (dysthymia, major depression, generalized anxiety disorders, antisocial personality disorder, nicotine dependence, and alcohol abuse/dependence). Logistic regression showed that nonsmoking for more than a year compared with less than 24 hr (nonabstinence) was significantly associated with reduced risk for wish to die (odds ratio [OR]: 0.56, 95% CI: 0.49-0.65), suicidal thoughts (OR: 0.54, 95% CI: 0.48-0.62), and attempted suicide (OR: 0.32, 95 % CI: 0.26-0.41). With adjustments for lifetime psychiatric disorders, duration of abstinence was no longer significantly associated with the SROs. CONCLUSIONS: In the sample of ever smokers with lifetime depressed mood, an apparent protective effect of increased duration of smoking abstinence on susceptibility to suicidal behavior was neutralized by the presence of psychiatric disorders. The causal direction of these relationships is unclear, and these cross-sectional findings need confirmation in future prospective studies. PMID- 21622495 TI - Young adults' interpretations of tobacco brands: implications for tobacco control. AB - INTRODUCTION: Marketers have long recognized the power and importance of branding, which creates aspirational attributes that increase products' attractiveness. Although brand imagery has traditionally been communicated via mass media, packaging's importance in promoting desirable brand-attribute associations has increased. Knowledge of how groups prone to smoking experimentation interpret tobacco branding would inform the debate over plain packaging currently occurring in many countries. METHODS: We conducted 12 group discussions and four in-depth interviews with 66 young adult smokers and nonsmokers of varying ethnicities from two larger New Zealand cities and one provincial city. Participants evaluated 10 familiar and unfamiliar tobacco brands using brand personality attributes and discussed the associations they had made. RESULTS: Participants ascribed very different images to different brands when exposed to the packaging alone, regardless of whether they had seen or heard of the brands before. Perceptual mapping of brands and image attributes highlighted how brand positions varied from older, more traditional, and male oriented to younger, feminine, and "cool." CONCLUSIONS: Our findings emphasize the continuing importance of tobacco branding as a promotion tool, even when communicated only by packaging. The ease with which packaging alone enabled young people to identify brand attributes and the desirable associations these connoted illustrate how tobacco packaging functions as advertising. The results support measures such as plain packaging of tobacco products to reduce exposure to these overt behavioral cues. PMID- 21622496 TI - Weight change over eight years in relation to alcohol consumption in a cohort of continuing smokers and quitters. AB - INTRODUCTION: Stopping smoking results in weight gain. Avoidance of alcohol is often advocated to reduce cues to smoking, but the effect of alcohol consumption on body weight is unclear. METHODS: We used regression models to examine weight change by baseline alcohol consumption in quitting and continuing smokers. Weight was measured at baseline and at 8 years, and weekly alcohol consumption was reported at baseline in participants from the Oxfordshire general practices nicotine patch/placebo trial. Of 698 smokers attempting to stop smoking, 85 were abstinent for 8 years and 613 continued to smoke. RESULTS: The association between baseline alcohol consumption and weight change depended upon smoking status (p for interaction = .019). In smokers, there was no association with weight change, 0.005 (95% CI: -0.037 to 0.056) kg per UK unit (U) (8 g of ethanol) consumed each week. This was unmodified by gender and baseline body mass index (BMI). In quitters, there was a negative association with weight change, 0.174 (95% CI: -0.315 to -0.034) kg per U consumed each week (unmodified by gender and baseline BMI). Quitters who consumed 14 U (112 g ethanol) a week weighed a mean 2.4 kg less than quitters who did not drink. CONCLUSIONS: Quitting smokers who drink more alcohol appear to gain less weight after quitting than those who do not drink. This is consistent across studies, it may be accounted for by unmeasured confounders or it may be that alcohol reduces weight gain. If alcohol reduces weight gain, the advice for quitting smokers must balance the benefits and hazards of alcohol consumption. However, there is currently insufficient evidence to advise quitters who drink little or no alcohol to increase consumption. PMID- 21622497 TI - Air contamination due to smoking in German restaurants, bars, and other venues- before and after the implementation of a partial smoking ban. AB - INTRODUCTION: The present study examined the reduction in exposure to tobacco smoke in German hospitality venues following the implementation of a partial smoking ban by measuring the indoor air concentration of PM(2.5) in 2005 and 2009, that is, before and after the legislation was implemented. METHODS: The concentration of respirable suspended particles (PM(2.5)) in the indoor air of German hospitality venues was measured using a laser photometer (AM510). The prelegislation sample from 2005 included 80 venues of which 58 could be revisited in 2009. After replenishment, the postlegislation sample consisted of 79 venues. RESULTS: Compared with the prelegislation measurement, the concentration of PM(2.5) in hospitality venues was reduced significantly after introduction of the smoke-free legislation. The median mass concentration of PM(2.5) was reduced by 87.1% in coffee bars, by 88.7% in restaurants, by 66.3% in bars, and by 90.8% in discotheques. Notably, legal exemptions to the smoking ban are an issue: At the postlegislation measurement in 2009, the mass concentrations of PM(2.5) were substantially higher in venues allowing smoking in the whole venue or in a designated smoking room than in completely smoke-free venues. CONCLUSIONS: The German smoke-free legislation significantly reduced the levels of respirable suspended particles in the indoor air of hospitality venues, benefiting the health of employees and patrons alike. But legal exemptions attenuated the effectiveness of the policy. PMID- 21622498 TI - Race/ethnicity moderates the effect of prepartum menthol cigarette use on postpartum smoking abstinence. AB - INTRODUCTION: Little is known about the influence of prepartum menthol cigarette use on postpartum smoking abstinence or how race/ethnicity might moderate this relationship. The current study addressed that gap by testing these relationships among racially/ethnically diverse women who quit smoking during pregnancy (N = 244; 33% African American, 31% Latina, 36% White). METHODS: Continuation ratio logit models were used to examine the effects of prepartum menthol cigarette use on biochemically confirmed, continuous abstinence through 26 weeks postpartum using an intent-to-treat approach. Analyses controlled for age, race/ethnicity, partner status, income, education, treatment, number of prequit cigarettes smoked per day, time to the first cigarette of the day, and time (Week 8 or 26 data collection timepoint). An additional model tested the moderating effects of race/ethnicity by including an interaction term. RESULTS: Prepartum menthol cigarette use was not significantly associated with postpartum smoking abstinence in the overall sample. However, the interaction between menthol use and race/ethnicity was significant (p = .02). Among White women, menthol use was associated with significantly lower odds of maintaining postpartum smoking abstinence (p = .03; odds ratio = .19 [.04-.89]), and the effect approached significance among African American women (p = .08). CONCLUSIONS: This study provides the first evidence that prepartum menthol cigarette use may increase the risk of postpartum smoking relapse among White, and possibly African American, women who quit smoking during or immediately before pregnancy. Results suggest that White and African American prepartum menthol users may require different or more intensive cessation services to aid in the maintenance of postpartum smoking abstinence. Replication with larger samples, and a focus on understanding the mechanisms that underlie these relationships, are warranted. PMID- 21622499 TI - Four in one polypill halves predicted cardiovascular risk, international study shows. PMID- 21622500 TI - Most African governments invest a third of what they promised on child health. PMID- 21622501 TI - Changes required to health bill will lead to delays, says Clegg. PMID- 21622502 TI - Women Peace Laureates urge protection for women in armed conflict. PMID- 21622503 TI - Fewer tests for cancer and rational use of chemotherapy can help reduce costs, say US doctors. PMID- 21622504 TI - Amputation and intraosseous access in infants. PMID- 21622505 TI - The assessment and management of insomnia in primary care. PMID- 21622506 TI - Congenital cataract. PMID- 21622507 TI - Research funders agree to increase free access to published research. PMID- 21622508 TI - Consumer groups call for decision on cancer risk of mobile phones to be delayed. PMID- 21622509 TI - Is there a risk in avoiding risk for younger patients with aortic valve disease? PMID- 21622510 TI - Commentary: Unintended consequences of governance. PMID- 21622511 TI - Commentary: Talking to patients about surgical innovations. PMID- 21622512 TI - Investigation of diarrhoea in a traveller just returned from India. PMID- 21622513 TI - Health reforms won't work without major changes, say King's Fund and BMA. PMID- 21622514 TI - NHS managers add voice to calls for changes to health bill. PMID- 21622515 TI - New guidelines aim to increase recognition and treatment of anxiety disorders. PMID- 21622516 TI - Treatment standards not commercial interests should drive commissioning, say surgeons. PMID- 21622517 TI - Global Fund pressures China to engage with civil society groups. PMID- 21622518 TI - Longitudinal analysis of sleep in relation to BMI and body fat in children: the FLAME study. AB - OBJECTIVES: To determine whether reduced sleep is associated with differences in body composition and the risk of becoming overweight in young children. DESIGN: Longitudinal study with repeated annual measurements. SETTING: Dunedin, New Zealand. PARTICIPANTS: 244 children recruited from a birth cohort and followed from age 3 to 7. MAIN OUTCOME MEASURES: Body mass index (BMI), fat mass (kg), and fat free mass (kg) measured with bioelectrical impedance; dual energy x ray absorptiometry; physical activity and sleep duration measured with accelerometry; dietary intake (fruit and vegetables, non-core foods), television viewing, and family factors (maternal BMI and education, birth weight, smoking during pregnancy) measured with questionnaire. RESULTS: After adjustment for multiple confounders, each additional hour of sleep at ages 3-5 was associated with a reduction in BMI of 0.48 (95% confidence interval 0.01 to 0.96) and a reduced risk of being overweight (BMI >= 85th centile) of 0.39 (0.24 to 0.63) at age 7. Further adjustment for BMI at age 3 strengthened these relations. These differences in BMI were explained by differences in fat mass index (-0.43, -0.82 to -0.03) more than by differences in fat free mass index (-0.21, -0.41 to 0.00). CONCLUSIONS: Young children who do not get enough sleep are at increased risk of becoming overweight, even after adjustment for initial weight status and multiple confounding factors. This weight gain is a result of increased fat deposition in both sexes rather than additional accumulation of fat free mass. PMID- 21622519 TI - Is prolonged lack of sleep associated with obesity? PMID- 21622520 TI - Cervical syphilitic spondylodiscitis associated with neurosyphilis. PMID- 21622521 TI - Relapsing polychondritis-associated meningitis and encephalitis: response to infliximab. PMID- 21622522 TI - New drugs beyond biologics in rheumatoid arthritis: the kinase inhibitors. AB - Orally available small molecule compounds have recently been developed for the treatment of RA, and inhibitors of signalling cascades, specifically inhibitors of kinases, have reached advanced stages of clinical development. The p38 mitogen activated protein kinase blockers have shown poor clinical response despite encouraging preclinical data. In contrast, inhibitors of the non-receptor tyrosine kinases, spleen tyrosine kinase and janus kinase 3, have demonstrated a significant clinical efficacy together with an acceptable safety profile. We herein present a review on published preclinical and clinical data on these new drugs. PMID- 21622523 TI - Laser Doppler perfusion imaging in systemic sclerosis impaired response to cold stimulation involves digits and hand dorsum. AB - OBJECTIVES: To assess by Laser Doppler perfusion imaging (LDPI) skin blood perfusion of hands in patients with SSc and primary RP (PRP) at baseline and after cold stimulation (CS). In SSc patients, the associations between skin perfusion and nailfold video capillaroscopy (NVC) patterns were also evaluated. METHODS: Forty patients with SSc, 38 patients with PRP and 32 healthy controls were recruited. Skin blood flow of the hands was detected by Lisca Laser Doppler Perfusion Imager at baseline and after CS. Further laser Doppler scanning was performed for each hand at 0 (T(1)), 3 (T(2)), 7 (T(3)) and 15 min (T(4)). RESULTS: Baseline mean perfusion is significantly (P < 0.000 l) lower in SSc patients than in healthy controls. In SSc patients, mean perfusion is reduced after CS (P < 0.0001) and skin flow recovery (significant difference between T(0) and T(4), P < 0.0001) is incomplete. In SSc patients with low vascular damage (early and active capillaroscopic groups), the abnormal microvascular response to CS involves only the digits, while the perfusion of hands dorsum is normal. With the progression of vascular damage (late capillaroscopic groups), the abnormal microvascular response to CS also appears in the hand dorsum skin. In PRP patients, baseline hand perfusion is very low and the skin flow recovery after CS is absent (P < 0.05). CONCLUSION: In early SSc, the thermoregulation of finger skin is impaired, but only in advanced stages of microangiopathy does the skin of the hand dorsum show a vasomotor control failure. PMID- 21622524 TI - Redundancy and specificity of multiple trigger factor chaperones in Desulfitobacteria. AB - The ribosome-bound trigger factor (TF) chaperone assists folding of newly synthesized polypeptides and participates in the assembly of macromolecular complexes. In the present study we showed that multiple distinct TF paralogues are present in genomes of Desulfitobacteria, a bacterial genus known for its ability to grow using organohalide respiration. Two full-length TF chaperones and at least one truncated TF (lacking the N-terminal ribosome-binding domain) were identified, the latter being systematically linked to clusters of reductive dehalogenase genes encoding the key enzymes in organohalide respiration. Using a well-characterized heterologous chaperone-deficient Escherichia coli strain lacking both TF and DnaK chaperones, we demonstrated that all three TF chaperones were functional in vivo, as judged by their ability to partially suppress bacterial growth defects and protein aggregation in the absence of both major E. coli chaperones. Next, we found that the N-terminal truncated TF-like protein PceT functions as a dedicated chaperone for the cognate reductive dehalogenase PceA by solubilizing and stabilizing it in the heterologous system. Finally, we showed that PceT specifically interacts with the twin-arginine signal peptide of PceA. Taken together, our data define PceT (and more generally the new RdhT family) as a class of TF-like chaperones involved in the maturation of proteins secreted by the twin-arginine translocation pathway. PMID- 21622525 TI - Genetic characterization of enzymes involved in the priming steps of oxytetracycline biosynthesis in Streptomyces rimosus. AB - Tetracyclines are clinically important aromatic polyketides whose biosynthesis is catalysed by bacterial type II polyketide synthases (PKSs). Tetracyclines are biosynthesized starting with an amide-containing malonamate starter unit and the resulting C-2 carboxyamide is critical for the antibiotic activities. In this work, we genetically verified that an amidotransferase, OxyD, and a thiolase, OxyP, are involved in the biosynthesis and incorporation of the starter unit. First, two mutations, R248T and D268N, were found to be present in OxyD* encoded in Streptomyces rimosus ATCC 13224, a strain that produces the acetate-primed 2 acetyl-2-decarboxyamido-oxytetracycline (ADOTC) instead of the malonamate-primed oxytetracycline (OTC). Homology modelling suggested that in particular D268N may inactivate OxyD. Complementation of S. rimosus ATCC 13224 with wild-type OxyD restored OTC biosynthesis, thereby confirming the essential role of OxyD in the synthesis of the amide starter unit. Second, using a series of knockout and complementation approaches, we demonstrated that OxyP is most likely involved in maintaining fidelity of the amide-priming process via hydrolysis of the competing acetate priming starter units. While the inactivation of OxyP does not eliminate OTC biosynthesis, the ratio of acetate-primed ADOTC to malonamate-primed OTC is significantly increased. This suggests that OxyP plays an ancillary role in OTC biosynthesis and is important for minimizing the levels of ADOTC, a shunt product that has much weaker antibiotic activities than OTC. PMID- 21622526 TI - Molecular typing methods for outbreak detection and surveillance of invasive disease caused by Neisseria meningitidis, Haemophilus influenzae and Streptococcus pneumoniae, a review. AB - Invasive disease caused by the encapsulated bacteria Neisseria meningitidis, Haemophilus influenzae and Streptococcus pneumoniae remains an important cause of morbidity and mortality worldwide, despite the introduction of successful conjugate polysaccharide vaccines that target disease-associated strains. In addition, resistance, or more accurately reduced susceptibility, to therapeutic antibiotics is spreading in populations of these organisms. There is therefore a continuing requirement for the surveillance of vaccine and non-vaccine antigens and antibiotic susceptibilities among isolates from invasive disease, which is only partially met by conventional methods. This need can be met with molecular and especially nucleotide sequence-based typing methods, which are fully developed in the case of N. meningitidis and which could be more widely deployed in clinical laboratories for S. pneumoniae and H. influenzae. PMID- 21622527 TI - Tannerella forsythia invasion in oral epithelial cells requires phosphoinositide 3-kinase activation and clathrin-mediated endocytosis. AB - Tannerella forsythia, a Gram-negative anaerobe implicated in periodontitis, has been detected within human buccal epithelial cells and shown to invade oral epithelial cells in vitro. We have previously shown that this bacterium triggers host tyrosine kinase-dependent phosphorylation and actin-dependent cytoskeleton reorganization for invasion. On the bacterial side, the leucine-rich repeat cell surface BspA protein is important for entry. The present study was undertaken to identify host signalling molecules during T. forsythia entry into human oral and cervical epithelial cells. Specifically, the roles of phosphatidylinositol 3 kinase (PI3K), Rho-family GTPases, cholesterol-rich membrane microdomains and the endocytic protein clathrin were investigated. For this purpose, cell lines were pretreated with chemical inhibitors or small interfering RNAs (siRNAs) that target PI3Ks, Rho GTPases, clathrin and cholesterol (a critical component of 'lipid rafts'), and the resulting effects on T. forsythia uptake were determined. Our studies revealed that T. forsythia entry is dependent on host PI3K signalling, and that purified BspA protein causes activation of this lipid kinase. Bacterial entry also requires the cooperation of host Rac1 GTPase. Finally, our findings indicate an important role for clathrin and cholesterol rich lipid microdomains in the internalization process. PMID- 21622528 TI - The autoregulator receptor homologue AvaR3 plays a regulatory role in antibiotic production, mycelial aggregation and colony development of Streptomyces avermitilis. AB - The gamma-butyrolactone autoregulator receptor has been shown to control secondary metabolism and/or morphological differentiation across many Streptomyces species. Streptomyces avermitilis produces an important anthelmintic agent (avermectin) and two further polyketide antibiotics, filipin and oligomycin. Genomic analysis of S. avermitilis revealed that this micro-organism has the clustered putative autoregulator receptor genes distant from the antibiotic biosynthetic gene clusters. Here, we describe the characterization of avaR3, one of the clustered receptor genes, which encodes a protein containing an extra stretch of amino acid residues that has not been found in the family of autoregulator receptors. Disruption of avaR3 resulted in markedly decreased production of avermectins, with delayed expression of avermectin biosynthetic genes, suggesting that AvaR3 positively controls the avermectin biosynthetic genes. Moreover, the disruption caused increased production of filipin without any changes in the transcriptional profile of the filipin biosynthetic genes, suggesting that filipin production is indirectly controlled by AvaR3. The avaR3 disruptant displayed fragmented growth in liquid culture and conditional morphological defects on solid medium. These findings demonstrated that AvaR3 acts as a global regulator that controls antibiotic production and cell morphology. PMID- 21622529 TI - Investigation of the population structure of Legionella pneumophila by analysis of tandem repeat copy number and internal sequence variation. AB - The population structure of the species Legionella pneumophila was investigated by multilocus variable number of tandem repeats (VNTR) analysis (MLVA) and sequencing of three VNTRs (Lpms01, Lpms04 and Lpms13) in selected strains. Of 150 isolates of diverse origins, 136 (86 %) were distributed into eight large MLVA clonal complexes (VACCs) and the rest were either unique or formed small clusters of up to two MLVA genotypes. In spite of the lower degree of genome-wide linkage disequilibrium of the MLVA loci compared with sequence-based typing, the clustering achieved by the two methods was highly congruent. The detailed analysis of VNTR Lpms04 alleles showed a very complex organization, with five different repeat unit lengths and a high level of internal variation. Within each MLVA-defined VACC, Lpms04 was endowed with a common recognizable pattern with some interesting exceptions. Evidence of recombination events was suggested by analysis of internal repeat variations at the two additional VNTR loci, Lpms01 and Lpms13. Sequence analysis of L. pneumophila VNTR locus Lpms04 alone provides a first-line assay for allocation of a new isolate within the L. pneumophila population structure and for epidemiological studies. PMID- 21622530 TI - Endocrine cells and blood vessels work in tandem to generate hormone pulses. AB - Hormones are dynamically collected by fenestrated capillaries to generate pulses, which are then decoded by target tissues to mount a biological response. To generate hormone pulses, endocrine systems have evolved mechanisms to tightly regulate blood perfusion and oxygenation, coordinate endocrine cell responses to secretory stimuli, and regulate hormone uptake from the perivascular space into the bloodstream. Based on recent findings, we review here the mechanisms that exist in endocrine systems to regulate blood flow, and facilitate coordinated cell activity and output under both normal physiological and pathological conditions in the pituitary gland and pancreas. PMID- 21622531 TI - Calcineurin (CN) activation promotes apoptosis of glomerular podocytes both in vitro and in vivo. AB - To determine the role of Gq signaling and calcineurin (CN) activation in promoting apoptosis of glomerular podocytes, constitutively active Gq [Gq(+)] or CN [CN(+)] proteins were introduced into cultured podocytes using protein transduction by tagging the proteins with the transactivator of transcription peptide. To investigate the role of CN in promoting podocyte apoptosis in vivo, a genetic model of type 1 diabetes mellitus (Akita mice) was treated with the CN inhibitor FK506. In cultured podocytes, Gq(+) stimulated nuclear translocation of nuclear factor of activated T cells (NFAT) family members, activated an NFAT reporter construct, and enhanced podocyte apoptosis in a CN-dependent fashion. CN(+) similarly promoted podocyte apoptosis, and apoptosis induced by either angiotensin II or endothelin-1 was blocked by FK506. Induction of apoptosis required NFAT-induced gene transcription because apoptosis induced by either Gq(+) or CN(+) was blocked by an inhibitor that prevented CN-dependent NFAT activation without affecting CN phosphatase activity. Podocyte apoptosis was mediated, in part, by the NFAT-responsive gene cyclooxygenase 2 (COX2) and prostaglandin E(2) generation because apoptosis induced by Gq(+) was attenuated by either COX2 inhibition or blockade of the Gq-coupled E-series prostaglandins receptor. The findings appeared relevant to podocyte apoptosis in diabetic nephropathy because apoptosis was significantly reduced in Akita mice by treatment with FK506. These data suggest that Gq stimulates CN and promotes podocyte apoptosis both in vitro and in vivo. Apoptosis requires NFAT-dependent gene transcription and is mediated, in part, by CN-dependent COX2 induction, prostaglandin E(2) generation, and autocrine activation of the Gq-coupled E series prostaglandins receptor. PMID- 21622532 TI - A mechanism for pituitary-resistance to thyroid hormone (PRTH) syndrome: a loss in cooperative coactivator contacts by thyroid hormone receptor (TR)beta2. AB - Thyroid hormone receptors (TR) are hormone-modulated transcription factors that regulate overall metabolic rate, lipid utilization, heart rate, and development. TR are expressed as a mix of interrelated receptor isoforms. The TRbeta2 isoform is expressed in the hypothalamus and pituitary, where it plays an important role in the feedback regulation of thyroid hormone levels. TRbeta2 exhibits unique transcriptional properties that parallel the ability of this isoform to bind to certain coactivators cooperatively through multiple contact surfaces. The more peripherally expressed TRbeta1 isoform, in contrast, appears to recruit these coactivators through a single contact mechanism. We report here that clusters of charged amino acids in the TR hormone-binding domain are required for this enhanced mode of coactivator recruitment and that mutations in these charge clusters, by disrupting TRbeta2 coactivator binding, are a molecular basis for pituitary resistance to thyroid hormone, a disease characterized by inappropriate thyroid hormone feedback regulation. We propose that the charge clusters allow wild-type TRbeta2 to assume a conformation compatible with its mode of multiple contact coactivator recruitment, whereas disruption of these charge clusters disrupts normal T(3) homeostasis by reducing TRbeta2 to a TRbeta1-like, single contact mode of coactivator binding. PMID- 21622533 TI - StarD7 gene expression in trophoblast cells: contribution of SF-1 and Wnt-beta catenin signaling. AB - Steroidogenic acute regulatory protein-related lipid transfer domain containing 7 (StarD7) is a poorly characterized member of the steroidogenic acute regulatory protein-related lipid transfer proteins, up-regulated in JEG-3 cells, involved in intracellular transport and metabolism of lipids. Previous studies dealing with the mechanisms underlying the human StarD7 gene expression led us to define the cis-acting regulatory sequences in the StarD7 promoter using as a model JEG-3 cells. These include a functional T cell-specific transcription factor 4 (TCF4) site involved in Wnt-beta-catenin signaling. To understand these mechanisms in more depth, we examined the steroidogenic factor 1 (SF-1) contribution to StarD7 expression. Cotransfection experiments in JEG-3 cells point out that the StarD7 promoter is activated by SF-1, and this effect is increased by forskolin. EMSA using JEG-3 nuclear proteins demonstrated that SF-1 binds to the StarD7 promoter. Additionally, chromatin immunoprecipitation analysis indicated that SF-1 and beta catenin are bound in vivo to the StarD7 promoter. Reporter gene assays in combination with mutations in the SF-1 and TCF4 binding sites revealed that the StarD7 promoter is synergistically activated by SF-1 and beta-catenin and that the TCF4 binding site (-614/-608) plays an important role in this activation. SF 1 amino acid mutations involved in the physical interaction with beta-catenin abolished this activation; thus demonstrating that the contact between the two proteins is necessary for an efficient StarD7 transcriptional induction. Finally, these data suggest that beta-catenin could function as a bridge between SF-1 and TCF4 forming a ternary complex, which would stimulate StarD7 expression. The SF-1 and beta-catenin pathway convergence on StarD7 expression may have important implications in the phospholipid uptake and transport, contributing to the normal trophoblast development. PMID- 21622534 TI - Mutant thyroid hormone receptors (TRs) isolated from distinct cancer types display distinct target gene specificities: a unique regulatory repertoire associated with two renal clear cell carcinomas. AB - Thyroid hormone receptors (TRs) are hormone-regulated transcription factors that regulate a diverse array of biological activities, including metabolism, homeostasis, and development. TRs also serve as tumor suppressors, and aberrant TR function (via mutation, deletion, or altered expression) is associated with a spectrum of both neoplastic and endocrine diseases. A particularly high frequency of TR mutations has been reported in renal clear cell carcinoma (RCCC) and in hepatocellular carcinoma (HCC). We have shown that HCC-TR mutants regulate only a fraction of the genes targeted by wild-type TRs but have gained the ability to regulate other, unique, targets. We have suggested that this altered gene recognition may contribute to the neoplastic phenotype. Here, to determine the generality of this phenomenon, we examined a distinct set of TR mutants associated with RCCC. We report that two different TR mutants, isolated from independent RCCC tumors, possess greatly expanded target gene specificities that extensively overlap one another, but only minimally overlap that of the wild-type TRs, or those of two HCC-TR mutants. Many of the genes targeted by either or both RCCC-TR mutants have been previously implicated in RCCC and include a series of metallothioneins, solute carriers, and genes involved in glycolysis and energy metabolism. We propose as a hypothesis that TR mutations from RCCC and HCC may play tissue-specific roles in carcinogenesis, and that the divergent target gene recognition patterns of TR mutants isolated from the two different types of tumors may arise from different selective pressures during development of RCCC vs. HCC. PMID- 21622535 TI - Mice lacking TR4 nuclear receptor develop mitochondrial myopathy with deficiency in complex I. AB - The estimated incidence of mitochondrial diseases in humans is approximately 1:5000 to 1:10,000, whereas the molecular mechanisms for more than 50% of human mitochondrial disease cases still remain unclear. Here we report that mice lacking testicular nuclear receptor 4 (TR4(-/-)) suffered mitochondrial myopathy, and histological examination of TR4(-/-) soleus muscle revealed abnormal mitochondrial accumulation. In addition, increased serum lactate levels, decreased mitochondrial ATP production, and decreased electron transport chain complex I activity were found in TR4(-/-) mice. Restoration of TR4 into TR4(-/-) myoblasts rescued mitochondrial ATP generation capacity and complex I activity. Further real-time PCR quantification and promoter studies found TR4 could modulate complex I activity via transcriptionally regulating the complex I assembly factor NDUFAF1, and restoration of NDUFAF1 level in TR4(-/-) myoblasts increased mitochondrial ATP generation capacity and complex I activity. Together, these results suggest that TR4 plays vital roles in mitochondrial function, which may help us to better understand the pathogenesis of mitochondrial myopathy, and targeting TR4 via its ligands/activators may allow us to develop better therapeutic approaches. PMID- 21622536 TI - The extreme C-terminal region of Galphas differentially couples to the luteinizing hormone and beta2-adrenergic receptors. AB - The mechanisms of G protein coupling to G protein-coupled receptors (GPCR) share general characteristics but may exhibit specific interactions unique for each GPCR/G protein partnership. The extreme C terminus (CT) of G protein alpha subunits has been shown to be important for association with GPCR. Hypothesizing that the extreme CT of Galpha(s) is an essential component of the molecular landscape of the GPCR, human LH receptor (LHR), and beta(2)-adrenergic receptor (beta(2)-AR), a model cell system was created for the expression and manipulation of Galpha(s) subunits in LHR(+) s49 ck cells that lack endogenous Galpha(s). On the basis of studies involving truncations, mutations, and chain extensions of Galpha(s), the CT was found to be necessary for LHR and beta(2)-AR signaling. Some general similarities were found for the responses of the two receptors, but significant differences were also noted. Computational modeling was performed with a combination of comparative modeling, molecular dynamics simulations, and rigid body docking. The resulting models, focused on the Galpha(s) CT, are supported by the experimental observations and are characterized by the interaction of the four extreme CT amino acid residues of Galpha(s) with residues in LHR and beta(2)-AR helix 3, (including R of the DRY motif), helix 6, and intracellular loop 2. This portion of Galpha(s) recognizes the same regions of the two GPCR, although with differences in the details of selected interactions. The predicted longer cytosolic extensions of helices 5 and 6 of beta(2)-AR are expected to contribute significantly to differences in Galpha(s) recognition by the two receptors. PMID- 21622537 TI - SMADs and FOXL2 synergistically regulate murine FSHbeta transcription via a conserved proximal promoter element. AB - Pituitary FSH regulates ovarian and testicular function. Activins stimulate FSHbeta subunit (Fshb) gene transcription in gonadotrope cells, the rate-limiting step in mature FSH synthesis. Activin A-induced murine Fshb gene transcription in immortalized gonadotropes is dependent on homolog of Drosophila mothers against decapentaplegic (SMAD) proteins as well as the forkhead transcription factor FOXL2 (FOXL2). Here, we demonstrate that FOXL2 synergizes with SMAD2, SMAD3, and SMAD4 to stimulate murine Fshb promoter-reporter activity in heterologous cells. Moreover, SMAD3-induction of Fshb promoter activity or endogenous mRNA expression is dependent upon endogenous FOXL2 in homologous cells. FOXL2/SMAD synergy requires binding of both FOXL2 and SMAD3 or SMAD4 to DNA. Of three putative forkhead-binding elements identified in the murine Fshb promoter, only the most proximal is absolutely required for activin A induction of reporter activity in homologous cells. Additionally, mutations to the minimal SMAD-binding element adjacent to the proximal forkhead-binding element abrogate activin A or FOXL2/SMAD3 induction of reporter activity. In contrast, a mutation that impairs an adjacent PBX1/PREP1 (pre-B cell leukemia transcription factor 1-PBX/knotted-1 homeobox-1) binding site does not alter activin A-stimulated promoter activity in homologous cells. Collectively, these and previous data suggest a model in which activins stimulate formation of FOXL2-SMAD2/3/4 complexes, which bind to the proximal murine Fshb promoter to stimulate its transcription. Within these complexes, FOXL2 and SMAD3 or SMAD4 bind to adjacent cis-elements, with SMAD3 brokering the physical interaction with FOXL2. Because this composite response element is highly conserved, this suggests a general mechanism whereby activins may regulate and/or modulate Fshb transcription in mammals. PMID- 21622538 TI - Oral artesunate-amodiaquine and artemether-lumefantrine in the treatment of uncomplicated hyperparasitaemic Plasmodium falciparum malaria in children. AB - The therapeutic efficacy, changes in haematocrit and declines in parasitaemias were evaluated in 56 children with uncomplicated falciparum hyperparasitaemia after oral artesunate-amodiaquine or artemether-lumefantrine. All children recovered clinically within 2 days and without progression to severe malaria. Falls in haematocrit in the first 3 days after treatment began were similar and <5%. Declines in parasitaemias were monoexponential with both treatments with an estimated half-life of 1 h. PMID- 21622539 TI - Skin injury following prolonged fluoroscopy: early and late appearances. PMID- 21622540 TI - The pattern of urinary copper excretion and its response to treatment in patients with Wilson's disease. AB - BACKGROUND: It is generally accepted that patients with Wilson's disease excrete excess copper in urine. However, there has been no study, on a large series of patients, as to whether there are differences in the rate of excretion at different stages of the disease or what changes may be expected after treatment. DESIGN: The present study follows from an analysis of the results of urinary copper excretion of 192 patients with Wilson's disease seen between 1955 and 2000. These patients were divided into three groups, pre-symptomatic, hepatic and neurological Wilson's disease. Patients were studied for basal pre-treatment, 24 h urinary copper excretion and for 6 h after a test dose of 500 mg penicillamine. The tests were repeated after approximately 1 and 2 years of chelation therapy with either penicillamine, or in a small minority of cases, trientine. RESULTS: The basal, pre-treatment copper excretion was the lowest in pre-symptomatic patients (207.93 ug/24 h) and the highest in the hepatic patients (465.75 ug/24 h). Those with neurological Wilson's disease gave an intermediate figure (305.58 ug/24 h). The response to penicillamine was the highest in the neurological patients and the lowest in the pre-symptomatic group. After 1 and 2 years of treatment all groups showed significant falls in both the basal and the after penicillamine rate of excretion of copper. The small subgroup treated with trientine, rather than penicillamine, showed similar results. CONCLUSIONS: The rate of copper excretion in patients with Wilson's disease shows wide variation from patient to patient, but in general patients with pre-symptomatic disease excrete less copper than those with symptomatic disease. All groups show a great increase when challenged with penicillamine. After 1 and 2 years of treatment, there is significant decrease in copper excretion in both basal and after penicillamine challenge. This presumably indicates a reduction in the body load of copper. PMID- 21622541 TI - Changes in NHS organization of care and management of hospital admissions with COPD exacerbations between the national COPD audits of 2003 and 2008. AB - BACKGROUND: The 2003 UK Chronic Obstructive Pulmonary Disease (COPD) audit revealed wide variability between hospital units in care delivered. AIMS: To assess whether processes of care, patient outcomes and organization of care have improved since 2003. DESIGN: A UK national audit was performed in 2008 to survey the organization and delivery of clinical care provided to patients admitted to hospital with COPD. METHODS: All UK acute hospital Trusts (units) were invited to participate. Each unit completed cross-sectional resource and organization questionnaires and a prospective clinical audit comprising up to 60 consecutively admitted cases of COPD exacerbation. Comparison between 2003 and 2008 includes aggregated statistics for units participating in both audit rounds. RESULTS: A total of 192 units participated in both audit rounds (6197 admissions in 2003 and 8170 in 2008). In 2008, patients were older and of a poorer functional class. Overall mortality was unchanged but adjusting for age and performance status, inpatient mortality (P = 0.05) and 90-day mortality (P = 0.001) were both reduced in 2008. More patients were discharged under a respiratory specialist (P < 0.01), treated with non-invasive ventilation if acidotic (P < 0.001) and accepted onto early discharge schemes (P < 0.01) while median length of stay fell from 6 to 5 days (P < 0.001). Within these mean data, however, there remains considerable inter-unit variation in organization, resources and outcomes. CONCLUSION: Overall improvements in resources and organization are accompanied by reduced mortality, shorter admissions and greater access to specialist services. There remains, however, considerable variation in the quality of secondary care provided between units. PMID- 21622542 TI - The role of energy-efficient biotechnological processes in the waste management industry. PMID- 21622543 TI - Learning transcriptional regulation on a genome scale: a theoretical analysis based on gene expression data. AB - The recent advent of high-throughput microarray data has enabled the global analysis of the transcriptome, driving the development and application of computational approaches to study transcriptional regulation on the genome scale, by reconstructing in silico the regulatory interactions of the gene network. Although there are many in-depth reviews of such 'reverse-engineering' methodologies, most have focused on the practical aspect of data mining, and few on the biological problem and the biological relevance of the methodology. Therefore, in this review, from a biological perspective, we used a set of yeast microarray data as a working example, to evaluate the fundamental assumptions implicit in associating transcription factor (TF)-target gene expression levels and estimating TFs' activity, and further explore cooperative models. Finally we confirm that the detailed transcription mechanism is overly-complex for expression data alone to reveal, nevertheless, future network reconstruction studies could benefit from the incorporation of context-specific information, the modeling of multiple layers of regulation (e.g. micro-RNA), or the development of approaches for context-dependent analysis, to uncover the mechanisms of gene regulation. PMID- 21622544 TI - Automated quantification of high-resolution CT scan findings in individuals at risk for pulmonary fibrosis. AB - BACKGROUND: Automated methods to quantify interstitial lung disease (ILD) on high resolution CT (HRCT) scans in people at risk for pulmonary fibrosis have not been developed and validated. METHODS: Cohorts with familial pulmonary fibrosis (n = 126) or rheumatoid arthritis with and without ILD (n = 86) were used to develop and validate a computer program capable of quantifying ILD on HRCT scans, which imaged the lungs semicontinuously from the apices to the lung bases during end inspiration in the prone position. This method uses segmentation, texture analysis, training, classification, and grading to score ILD. RESULTS: Quantification of HRCT scan findings of ILD using an automated computer program correlated with radiologist readings and detected disease of varying severity in a derivation cohort with familial pulmonary fibrosis or their first-degree relatives. This algorithm was validated in an independent cohort of subjects with rheumatoid arthritis with and without ILD. Automated classification of HRCT scans as normal or ILD was significant in the derivation and validation cohorts (P < .001 and P < .001, respectively). Areas under receiver operating characteristic curves performed independently for each group were 0.888 for the derivation cohort and 0.885 for the validation cohort. Pulmonary function test results, including FVC and diffusion capacity, correlated with computer-generated HRCT scan scores for ILD (r = -0.483 and r = -0.532, respectively). CONCLUSIONS: Automated computer scoring of HRCT scans can objectively identify ILD and potentially quantify radiographic severity of lung disease in populations at risk for pulmonary fibrosis. PMID- 21622545 TI - Does the outcome justify an oral-first treatment strategy for management of pulmonary arterial hypertension? AB - BACKGROUND: Guidelines for the treatment of World Health Organization (WHO) functional class (FC) III pulmonary arterial hypertension (PAH) provide for oral (PO) therapy or parenteral prostacyclins at the discretion of expert physicians. The objective of this study was to assess the appropriateness of PO-first treatment in patients with WHO FC III PAH. METHODS: This study was a retrospective analysis of 79 treatment-naive adult patients with idiopathic, familial, or anorexigen-associated PAH, referred to a single pulmonary hypertension center. Forty-eight received either PO therapy with an endothelin receptor antagonist, calcium channel blocker, or phosphodiesterase inhibitor (PO group) and 31 an IV or subcutaneous (SC) prostacyclin (IV/SC group). RESULTS: Patients in the IV/SC group had a significantly worse baseline hemodynamic profile; however, on univariate analysis, there was no association between hemodynamics and mortality. Initial treatment with PO vs IV/SC therapy was associated with a lower overall mortality (20.8% vs 45.2%, P = .02) and a lower 5 year mortality (14.6% vs 32.3%, P = .062). Based on the National Institutes of Health equation, actual survival for patients who received PO therapy was greater than predicted at 5 years. Finally, there were similar improvements between groups in 6-min walk distance (P = .38) at 6 to 12 months after initiation of treatment. CONCLUSIONS: For WHO FC III PAH that is idiopathic, familial, or anorexigen associated, the clinical decision for treatment with a PO-first strategy is associated with a high survival rate when patients are appropriately risk stratified prior to initiation of therapy. The more potent prostacyclins can be reserved for high-risk patients, those with evidence of disease progression, or those with treatment failure. PMID- 21622546 TI - Classification of non-small cell lung carcinoma in transthoracic needle specimens using microRNA expression profiling. AB - BACKGROUND: Emerging targeted lung cancer therapies require the accurate morphologic subclassification of non-small cell lung cancer (NSCLC), even in scant and distorted specimens obtained by transthoracic needle aspiration (TTNA). MicroRNAs (miRNAs) are small noncoding genes recently reported as useful in differentiating squamous cell carcinoma (SCC) from adenocarcinoma (AD) in resected tumor specimens. We investigated their ability to do so in TTNA specimens. METHODS: Smears, immunocytochemistry slides, and corresponding cell blocks of 31 NSCLC TTNA specimens were retrieved and classified as AD or SCC based on their cytologic features and immunocytochemical profiles. Data on EGFR and K-RAS mutational status were available for all cases of AD. We quantified the hsa-let-7 family and hsa-miR-205 by quantitative reverse transcription-polymerase chain reaction and compared the miRNA expression levels in AD and SCC using Student t test. RESULTS: Eighteen cases were classified as AD and 13 as SCC by light microscopy and immunocytochemistry. miRNA expression profiles demonstrated considerable, statistically significant differences between AD and SCC, showing an upregulation of hsa-let-7a, hsa-let-7b, hsa-let-7c,hsa-let-7f, hsa-let-7g, hsa let-7i, and hsa-miR-98 and a downregulation of hsa-miR-205 in AD specimens (all P < .05; t test). CONCLUSIONS: Profiling the hsa-let-7 family and hsa-miR-205 is a promising method for differentiating AD from SCC, even in such small specimens as transthoracic aspirates. Subject to the validation of these findings in further, larger studies, this could prove to be a reliable, standardizable tool for the subclassification of NSCLC. PMID- 21622547 TI - End tidal CO(2) tension: pulmonary arterial hypertension vs pulmonary venous hypertension and response to treatment. AB - BACKGROUND: CO(2) excretion is impaired in pulmonary arterial hypertension (PAH) due to underlying vascular obstruction and increased dead space. Our aim was to determine whether resting end tidal CO(2) (Etco(2)) could differentiate patients with PAH from those with pulmonary venous hypertension (PVH) or patients without pulmonary hypertension (PH) and whether successful treatment of PAH resulted in higher Etco(2) values. METHODS: We performed Etco(2) measurements for five breaths at rest and after a 6-min walk test (6MWT) in patients seen at our pulmonary vascular center. Mean Etco(2) values were correlated with 6-min walk distance and right-sided heart catheterization data. RESULTS: We enrolled 84 patients with PAH, 17 with PVH without left ventricular systolic dysfunction, and seven with no PH and no severe alterations in pulmonary function testing. Etco(2) was significantly lower in patients with PAH than in those with no PH and PVH (P < .0001 PAH vs both groups). Etco(2) correlated with the pulmonary artery diastolic pressure-to-pulmonary artery occlusion pressure gradient (r = -0.50, P = .0002) and pulmonary vascular resistance (r = -0.44, P = .002). Etco(2) after 6MWT correlated with walk distance (r = 0.34, P = .003). In patients with prostaglandin therapy escalation, Etco(2) increased in those who had clinical improvement, whereas in patients who did not improve clinically, Etco(2) failed to rise (P = .04). CONCLUSIONS: Etco(2) is a promising tool to differentiate patients with PAH from those with PVH or no PH, correlates with diagnostic and prognostic hemodynamic indicators, and may increase with successful treatment of PAH. PMID- 21622548 TI - Risk factors for tumor recurrence in patients with early-stage (stage I and II) non-small cell lung cancer: patient selection criteria for adjuvant chemotherapy according to the seventh edition TNM classification. AB - OBJECTIVES: The purpose of this study was to evaluate risk factors for tumor recurrence in patients with completely resected early-stage non-small cell lung cancer (NSCLC). METHODS: Between July 1992 and December 2007, 1,967 consecutive patients with stage I and II NSCLC with diagnoses based on the seventh edition TNM classification underwent complete resection. All patients were divided into three groups according to the stage and presence of lymph node metastasis: stage I, patients with stage I, T1-T2aN0M0 disease; stage IIN0, patients with stage II, T2b-T3N0M0, node-negative disease; and stage IIN1, patients with stage II, T1 2N1M0, node-positive disease. Freedom from recurrence rate was estimated using the Kaplan-Meier method, and recurrence risk factors were identified by univariate and multivariate analyses. RESULTS: The 5-year freedom from recurrence rates for stage I, stage IIN0, and stage IIN1 patients were 84%, 61%, and 54%, respectively. By multivariate analyses, three variables (histologic differentiation, vessel invasion, and visceral pleural invasion) in stage I and two variables (adenocarcinoma histology and visceral pleural invasion) in stage IIN0 and stage IIN1 were shown to be independently significant risk factors for recurrence. According to subgroup analyses that combined these risk factors in each group, the 5-year freedom from recurrence rate was 63% for stage I with three risk factors, whereas those for stage IIN0 and stage IIN1 without risk factors were 83% and 78%, respectively. CONCLUSION: In patients with stage I and II NSCLC, we identified risk factors for recurrence. When these factors are combined, high- and low-risk subgroups can be identified within each group. PMID- 21622549 TI - Persistence of community-acquired respiratory distress syndrome toxin-producing Mycoplasma pneumoniae in refractory asthma. AB - BACKGROUND: The role of Mycoplasma pneumoniae (Mp) in the initiation and persistence of asthma remains elusive. Mp community-acquired respiratory distress syndrome toxin (CARDS Tx) is a unique virulence factor that induces an intense lymphocytic response and exacerbates asthma in animal models. We sought to determine the incidence of Mp infection and the presence of CARDS Tx in subjects with refractory asthma (RA). METHODS: We conducted a prospective observational study in 64 subjects with RA. Respiratory secretions (sputum, nasal lavage, and throat swab) and blood were analyzed for the presence of CARDS Tx and P1 adhesin (P1) DNA by polymerase chain reaction (PCR), and CARDS Tx by antigen capture. Serum IgM and IgG antibodies to CARDS Tx were determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: Thirty-three of 64 subjects (52%) tested positive for Mp: 29 of 33 by CARDS Tx vs 10 of 33 by P1 assays. Ten subjects followed longitudinally for up to 633 days tested persistently positive for Mp. There were no significant differences in Mp-specific IgG responses between Mp positive and Mp-negative groups. Eight of 10 subjects who tested persistently positive failed to mount a substantial IgG response to CARDS Tx, and up to 8 weeks of clarithromycin failed to eradicate Mp in five subjects. CONCLUSIONS: Subjects with RA may be chronically infected with Mp. PCR for CARDS Tx appears to be the most sensitive method of identifying Mp infection. Despite the persistence of Mp in subjects with RA, some subjects failed to mount an IgG response, and macrolide therapy was insufficient to eradicate Mp. PMID- 21622550 TI - Lung MRI as a possible alternative to CT scan for patients with primary immune deficiencies and increased radiosensitivity. AB - BACKGROUND: Patients with common variable immunodeficiency (CVID) suffer from respiratory infections leading over time to permanent lung damage. Increased radiosensitivity has been described, and clinicians should consider a risk benefit assessment when ordering a CT scan, in that the exact level of "safe" radiation exposure is unknown. METHODS: Twenty-one patients with CVID were evaluated with chest CT scan, MRI, and pulmonary function tests on the same day. MRI protocol included a T2-weighted rotating blade-like k-space covering sequence (time repetition, 2,000; echo train = 27; field of view, 400 mm; flip angle, 150; slice thickness, 5 mm) on axial and coronal planes. The bronchial and parenchymal abnormalities were compared with those identified by CT scan applying a modified Bhalla scoring system to assess bronchiectasis, bronchial wall thickening, number of bronchial generations involved, mucous plugging, consolidations, emphysema, bullae, and nodules. RESULTS: CT scan and MRI findings were comparable for moderate to severe degrees of bronchial and parenchymal alterations. A low concordance was found between MRI and CT scan for lower scores of bronchial abnormalities. CT scan allowed a better identification of peripheral airways abnormalities. CONCLUSIONS: Lung alterations in patients with higher radiation sensitivity, such as patients with CVID, might be evaluated by MRI, a radiation free technique alternative to CT scan. PMID- 21622551 TI - Intramodality and intermodality comparisons of storage phosphor computed radiography and conventional film-screen radiography in the recognition of small pneumoconiotic opacities. AB - BACKGROUND: Digital radiography systems are replacing traditional film for chest radiographic monitoring in the recognition of pneumoconiosis. METHODS: To further investigate previous findings regarding the equivalence of film-screen radiographs (FSRs) and storage phosphor computed radiographs (CRs), FSRs and CRs from 172 underground coal miners were classified independently by seven National Institute for Occupational Safety and Health-approved B readers, using the International Labor Office (ILO) classification of radiographs of pneumoconiosis. RESULTS: More CRs were classified as "good" quality compared with FSRs (prevalence ratio [PR], 1.5; 95% CI, 1.4-1.6; P , .001). B readers showed good overall agreement on scoring small opacity profusion using CRs vs FSRs (weighted kappa, 0.58; 95% CI, 0.54-0.62). Significantly more irregular opacities (compared with rounded) were classified using CR images compared with FSR (PR, 1.3; 95% CI, 1.1-1.6; P = .01). Similarly, the smallest sized opacities (width < 1.5 mm, p and s type) were reported more frequently using CR vs FSR images (PR, 1.3; 95% CI, 1.1-1.5; P < .001). Interreader and intrareader agreement was lower with respect to the classification of shape and size than for small opacity profusion. Overall, interreader and intrareader variability did not differ significantly using CR vs FSR. CONCLUSIONS: Under optimal conditions, using standardized methods and equipment, reader visualization of small pneumoconiotic opacities does not appear to differ meaningfully, whether using CR or FSR. Variability in ILO classifications between imaging modalities appears to be considerably lower than variability among readers. The well-documented challenge of reader variability does not appear to be resolved through the use of digital imaging alone, and additional approaches must be evaluated. PMID- 21622552 TI - Does propulsion mechanism influence the long-term side effects of oral appliances in the treatment of sleep-disordered breathing? AB - BACKGROUND: Mandibular advancement appliances (MAAs) can be used to treat sleep disordered breathing, and differences in their designs have been thought to influence the occurrence of secondary effects. With bibloc devices, the lateral attachments producing mandibular propulsion can be inserted to generate traction or compression. We evaluated the clinical impact of this difference by comparing the long-term secondary effects and compliance of two devices. METHODS: One hundred sixty-two records of patients fitted with a traction-based or compression based MAA before January 2008 were reviewed retrospectively for physical examination findings and cephalometry. Patients were sent a postal survey and contacted by phone 2 weeks later. They were offered a follow-up medical visit with repeat cephalometry. Main outcomes were long-term (> 2 years) secondary effects, compliance, or satisfaction. RESULTS: Of the patients who attended the follow-up visit, 48 had worn the MAA for > 2 years (16 traction based and 32 compression based), and nine for < 6 months (used as control subjects). Mean follow-up times for the traction and compression groups were 3.7 +/- 1.2 years and 3.6 +/- 1.2 years, respectively. No difference was found between the MAAs for subjective and objective side effects, except for greater early pain to the masticatory muscles (P = .02) and residual tongue pain (P = .04) in the compression group. However, pain intensity was low and clinical relevance was uncertain. No difference was found for compliance, satisfaction, and objective or subjective efficacy. CONCLUSIONS: This study suggests that traction-based appliances are similar to compression-based devices with regard to secondary effects and compliance. PMID- 21622553 TI - A 12-year follow-up study of patients with newly diagnosed lone atrial fibrillation: implications of arrhythmia progression on prognosis: the Belgrade Atrial Fibrillation study. AB - BACKGROUND: Lone atrial fibrillation (AF) has been suggested to have a favorable long-term prognosis. Significant interest has been directed at factors predicting arrhythmia progression, and the HATCH score (hypertension, age >= 75 years, transient ischemic attack or stroke [2 points], COPD, and heart failure [2 points]) recently has been proposed as a predictive score for AF progression. We investigated long-term outcomes in a large cohort of newly diagnosed lone AF and whether progression from paroxysmal to permanent AF confers an adverse impact on outcomes, including stroke and thromboembolism. METHODS: The study was an observational cohort of 346 patients with newly diagnosed lone AF with a mean follow-up of 12.1 +/- 7.3 years. RESULTS: Baseline paroxysmal AF was confirmed in 242 patients, and of these, 65 (26.9%) subsequently experienced progression to permanent AF. Older age and development of congestive heart failure during follow up were the multivariate predictors of AF progression (both P < .01), which was documented in 19.8% of patients with a HATCH score of 0 vs 63.2% with a score of 2 (P < .001), although the predictive validity of the HATCH score per se was modest (C statistic, 0.6). The annual rate of thromboembolism and heart failure during follow-up were low (0.4% each), and five patients (1.4%) died. AF progression, development of cardiac diseases, and older age were multivariate predictors of adverse outcomes, including thromboembolism (all P < .05). Baseline CHADS(2) (congestive heart failure, hypertension, age >= 75, diabetes mellitus, prior stroke or transient ischemic attack) score was not predictive for thromboembolism (C statistic, 0.50; 95% CI, 0.31-0.69). CONCLUSIONS: This 12-year follow-up study provides confirmatory evidence of a generally favorable prognosis of lone AF, but adverse outcomes (including stroke and thromboembolism) are significantly influenced by age and the (new) development of underlying heart disease. Arrhythmia progression in lone AF is a marker of increased risk for adverse cardiovascular events. PMID- 21622554 TI - Retroviral integrases promote fraying of viral DNA ends. AB - In the initial step of integration, retroviral integrase (IN) introduces precise nicks in the degenerate, short inverted repeats at the ends of linear viral DNA. The scissile phosphodiester bond is located immediately 3' of a highly conserved CA/GT dinucleotide, usually 2 bp from the ends. These nicks create new recessed 3'-OH viral DNA ends that are required for joining to host cell DNA. Previous studies have indicated that unpairing, "fraying," of the viral DNA ends by IN contributes to end recognition or catalysis. Here, we report that end fraying can be detected independently of catalysis with both avian sarcoma virus (ASV) and human immunodeficiency virus type 1 (HIV-1) IN proteins by use of fluorescence resonance energy transfer (FRET). The results were indicative of an IN-induced intramolecular conformational change in the viral DNA ends (cis FRET). Fraying activity is tightly coupled to the DNA binding capabilities of these enzymes, as follows: an inhibitor effective against both IN proteins was shown to block ASV IN DNA binding and end fraying, with similar dose responses; ASV IN substitutions that reduced DNA binding also reduced end fraying activity; and HIV-1 IN DNA binding and end fraying were both undetectable in the absence of a metal cofactor. Consistent with our previous results, end fraying is sequence independent, suggesting that the DNA terminus per se is a major structural determinant for recognition. We conclude that frayed ends represent a functional intermediate in which DNA termini can be sampled for suitability for endonucleolytic processing. PMID- 21622555 TI - Structural and functional analyses reveal that Staphylococcus aureus antibiotic resistance factor HmrA is a zinc-dependent endopeptidase. AB - HmrA is an antibiotic resistance factor of methicillin-resistant Staphylococcus aureus. Molecular analysis of this protein revealed that it is not a muramidase or beta-lactamase but a nonspecific double-zinc endopeptidase consisting of a catalytic domain and an inserted oligomerization domain, which probably undergo a relative interdomain hinge rotation upon substrate binding. The active-site cleft is located at the domain interface. Four HmrA protomers assemble to a large ~170 kDa homotetrameric complex of 125 A. All four active sites are fully accessible and ~50-70 A apart, far enough apart to act on a large meshwork substrate independently but simultaneously. In vivo studies with four S. aureus strains of variable resistance levels revealed that the extracellular addition of HmrA protects against loss of viability in the presence of oxacillin and that this protection depends on proteolytic activity. All of these results indicate that HmrA is a peptidase that participates in resistance mechanisms in vivo in the presence of beta-lactams. Furthermore, our results have implications for most S. aureus strains of known genomic sequences and several other cocci and bacilli, which harbor close orthologs. This suggests that HmrA may be a new widespread antibiotic resistance factor in bacteria. PMID- 21622556 TI - Receptor-like protein-tyrosine phosphatase alpha enhances cell surface expression of neural adhesion molecule NB-3. AB - Neural adhesion molecule NB-3 plays an important role in the apical dendrite development of layer V pyramidal neurons in the visual cortex, and receptor-like protein-tyrosine phosphatase alpha (PTPalpha) mediates NB-3 signaling in this process. Here we investigated the role of PTPalpha in regulating cell surface expression of NB-3. We found that cortical neurons from PTPalpha knock-out mice exhibited a lower level of NB-3 at the cell surface. When expressed in COS1 cells, NB-3 was enriched in the Golgi apparatus with a low level of cell surface expression. However, co-expression of PTPalpha increased the cell surface distribution of NB-3. Further analysis showed that PTPalpha facilitated Golgi exit of NB-3 and stabilized NB-3 protein at the cell surface by preventing its release from the plasma membrane. The extracellular region of PTPalpha but not its catalytic activity is necessary for its effect on NB-3 expression. Thus, the PTPalpha-mediated increase of NB-3 level at the cell surface represents a novel function of PTPalpha in NB-3 signaling in neural development. PMID- 21622557 TI - Cellular mechanisms for dopamine D4 receptor-induced homeostatic regulation of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors. AB - Aberrant dopamine D(4) receptor function has been implicated in mental illnesses, including schizophrenia and attention deficit-hyperactivity disorder. Recently we have found that D(4) receptor exerts an activity-dependent bi-directional regulation of AMPA receptor (AMPAR)-mediated synaptic currents in pyramidal neurons of prefrontal cortex (PFC) via the dual control of calcium/calmodulin kinase II (CaMKII) activity. In this study, we examined the signaling mechanisms downstream of CaMKII that govern the complex effects of D(4) on glutamatergic transmission. We found that in PFC neurons at high activity state, D(4) suppresses AMPAR responses by disrupting the kinesin motor-based transport of GluR2 along microtubules, which was accompanied by the D(4) reduction of microtubule stability via a mechanism dependent on CaMKII inhibition. On the other hand, in PFC neurons at the low activity state, D(4) potentiates AMPAR responses by facilitating synaptic targeting of GluR1 through the scaffold protein SAP97 via a mechanism dependent on CaMKII stimulation. Taken together, these results have identified distinct signaling mechanisms underlying the homeostatic regulation of glutamatergic transmission by D(4) receptors, which may be important for cognitive and emotional processes in which dopamine is involved. PMID- 21622558 TI - Oxygen supply from the bird's eye perspective: globin E is a respiratory protein in the chicken retina. AB - The visual process in the vertebrate eye requires high amounts of metabolic energy and thus oxygen. Oxygen supply of the avian retina is a challenging task because birds have large eyes, thick retinae, and high metabolic rates but neither deep retinal nor superficial capillaries. Respiratory proteins such as myoglobin may enhance oxygen supply to certain tissues, and thus the mammalian retina harbors high amounts of neuroglobin. Globin E (GbE) was recently identified as an eye-specific globin of chicken (Gallus gallus). Orthologous GbE genes were found in zebra finch and turkey genomes but appear to be absent in non avian vertebrate classes. Analyses of globin phylogeny and gene synteny showed an ancient origin of GbE but did not help to assign it to any specific globin type. We show that the photoreceptor cells of the chicken retina have a high level of GbE protein, which accumulates to ~10 MUM in the total eye. Quantitative real time RT-PCR revealed an ~50,000-fold higher level of GbE mRNA in the eye than in the brain. Spectroscopic analysis and ligand binding kinetics of recombinant chicken GbE reveal a penta-coordinated globin with an oxygen affinity of P(50) = 5.8 torrs at 25 degrees C and 15 torrs at 41 degrees C. Together these data suggest that GbE helps to sustain oxygen supply to the avian retina. PMID- 21622559 TI - Helix 69 is key for uniformity during substrate selection on the ribosome. AB - Structural studies of ribosome complexes with bound tRNAs and release factors show considerable contacts between these factors and helix 69 (H69) of 23 S rRNA. Although biochemical and genetic studies have provided some general insights into the role of H69 in tRNA and RF selection, a detailed understanding of these contributions remains elusive. Here, we present a pre- steady-state kinetic analysis establishing that two distinct regions of H69 make critical contributions to substrate selection. The loop of H69 (A1913) forms contacts necessary for the efficient accommodation of a subset of natural tRNA species, whereas the base of the stem (G1922) is specifically critical for UGA codon recognition by the class 1 release factor RF2. These data define a broad and critical role for this centrally located intersubunit helix (H69) in accurate and efficient substrate recognition by the ribosome. PMID- 21622560 TI - Detoxication of structurally diverse polycyclic aromatic hydrocarbon (PAH) o quinones by human recombinant catechol-O-methyltransferase (COMT) via O methylation of PAH catechols. AB - Polycyclic aromatic hydrocarbons (PAH) are environmental and tobacco carcinogens. Metabolic activation of intermediate PAH trans-dihydrodiols by aldo-keto reductases (AKRs) leads to the formation of electrophilic and redox-active o quinones. We investigated whether O-methylation by human recombinant soluble catechol-O-methyltransferase (S-COMT) is a feasible detoxication step for a panel of structurally diverse PAH-catechols produced during the redox-cycling process. Classes of PAH non-K-region o-quinones (bay region, methylated bay region, and fjord region o-quinones) produced by AKRs were employed in the studies. PAH o quinones were reduced to the corresponding catechols by dithiothreitol under anaerobic conditions and then further O-methylated by human S-COMT in the presence of S-[3H]adenosyl-l-methionine as a methyl group donor. The formation of the O-methylated catechols was detected by HPLC-UV coupled with in-line radiometric detection, and unlabeled products were also characterized by LC MS/MS. Human S-COMT was able to catalyze O-methylation of all of the PAH catechols and generated two isomeric metabolites in different proportions. LC MS/MS showed that each isomer was a mono-O-methylated metabolite. 1H NMR was used to assign the predominant positional isomer of benzo[a]pyrene-7,8-catechol as the O-8-monomethylated catechol. The catalytic efficiency (k(cat)/K(m)) varied among different classes of PAH-catechols by 500-fold. The ability of S-COMT to produce two isomeric products from PAH-catechols was rationalized using the crystal structure of the enzyme. We provide evidence that O-8-monomethylated benzo[a]pyrene-7,8-catechol is formed in three different human lung cell lines. It is concluded that human S-COMT may play a critical role in the detoxication of PAH o-quinones generated by AKRs. PMID- 21622561 TI - Internal polyadenylation of the parvovirus B19 precursor mRNA is regulated by alternative splicing. AB - Alternative processing of parvovirus B19 (B19V) pre-mRNA is critical to generating appropriate levels of B19V mRNA transcripts encoding capsid proteins and small nonstructural proteins. Polyadenylation of the B19V pre-mRNA at the proximal polyadenylation site ((pA)p), which prevents generation of full-length capsid proteins encoding mRNA transcripts, has been suggested as a step that blocks B19V permissiveness. We report here that efficient splicing of the B19V pre-mRNA within the first intron (upstream of the (pA)p site) stimulated the polyadenylation; in contrast, splicing of the B19V pre-mRNA within the second intron (in which the (pA)p site resides) interfered with the polyadenylation, leading to the generation of a sufficient number of B19V mRNA transcripts polyadenylated at the distal polyadenylation site ((pA)d). We also found that splicing within the second intron and polyadenylation at the (pA)p site compete during processing of the B19V pre-mRNA. Furthermore, we discovered that the U1 RNA that binds to the 5' splice donor site of the second intron is fully responsible for inhibiting polyadenylation at the (pA)p site, whereas actual splicing, and perhaps assembly of the functional spliceosome, is not required. Finally, we demonstrated that inhibition of B19V pre-mRNA splicing within the second intron by targeting an intronic splicing enhancer using a Morpholino antisense oligonucleotide prevented B19V mRNA transcripts polyadenylated at the (pA)d site during B19V infection of human erythroid progenitors. Thus, our study reveals the mechanism by which alternative splicing coordinates alternative polyadenylation to generate full-length B19V mRNA transcripts at levels sufficient to support productive B19V infection. PMID- 21622562 TI - Silencing suppressor of cytokine signaling-1 (SOCS1) in macrophages improves Mycobacterium tuberculosis control in an interferon-gamma (IFN-gamma)-dependent manner. AB - Protection against infection with Mycobacterium tuberculosis demands IFN-gamma. SOCS1 has been shown to inhibit responses to IFN-gamma and might thereby play a central role in the outcome of infection. We found that M. tuberculosis is a highly efficient stimulator of SOCS1 expression in murine and human macrophages and in tissues from infected mice. Surprisingly, SOCS1 reduced responses to IL 12, resulting in an impaired IFN-gamma secretion by macrophages that in turn accounted for a deteriorated intracellular mycobacterial control. Despite SOCS1 expression, mycobacteria-infected macrophages responded to exogenously added IFN gamma. SOCS1 attenuated the expression of the majority of genes modulated by M. tuberculosis infection of macrophages. Using a conditional knockdown strategy in mice, we found that SOCS1 expression by macrophages hampered M. tuberculosis clearance early after infection in vivo in an IFN-gamma-dependent manner. On the other hand, at later time points, SOCS1 expression by non-macrophage cells protected the host from infection-induced detrimental inflammation. PMID- 21622563 TI - Novel mechanism of anti-apoptotic function of 78-kDa glucose-regulated protein (GRP78): endocrine resistance factor in breast cancer, through release of B-cell lymphoma 2 (BCL-2) from BCL-2-interacting killer (BIK). AB - Activation of the intrinsic apoptotic pathway represents a major mechanism for breast cancer regression resulting from anti-estrogen therapy. The BH3-only protein BIK is inducible by estrogen-starvation and anti-estrogen treatment and plays an important role in anti-estrogen induced apoptosis of breast cancer cells. BIK is predominantly localized to the endoplasmic reticulum where it regulates BAX/BAK-dependent release of Ca(2+) from the endoplasmic reticulum stores and cooperates with other BH3-only proteins such as NOXA to cause rapid release of cytochrome c from mitochondria and activate apoptosis. BIK is also known to inactivate BCL-2 through complex formation. Previously, we demonstrated that apoptosis triggered by BIK in estrogen-starved human breast cancer cells is suppressed by GRP78, a major endoplasmic reticulum chaperone. Here we described the isolation of a novel clonal human breast cancer cell line (MCF-7/BUS-10) resistant to long-term estrogen deprivation. These cells exhibit elevated level of GRP78, which protects them from estrogen starvation-induced apoptosis. Our studies revealed that overexpression of GRP78 suppresses apoptosis induced by BIK and NOXA, either alone or in combination. Surprisingly, the interaction of GRP78 with BIK does not require its BH3 domain, which has been implicated in all previous BIK protein interactions. We further showed GRP78 and BCL-2 form independent complex with BIK and that increased expression of GRP78 decreases BIK binding to BCL-2. Our findings provide the first evidence that GRP78 can decrease BCL-2 sequestration by BIK at the endoplasmic reticulum, thus uncovering a potential new mechanism whereby GRP78 confers endocrine resistance in breast cancer. PMID- 21622564 TI - Regulatory roles of heterogeneous nuclear ribonucleoprotein M and Nova-1 protein in alternative splicing of dopamine D2 receptor pre-mRNA. AB - The dopamine D2 receptor (D2R) plays a crucial role in the regulation of diverse key physiological functions, including motor control, reward, learning, and memory. This receptor is present in vivo in two isoforms, D2L and D2S, generated from the same gene by alternative pre-mRNA splicing. Each isoform has a specific role in vivo, underlining the importance of a strict control of its synthesis, yet the molecular mechanism modulating alternative D2R pre-mRNA splicing has not been completely elucidated. Here, we identify heterogeneous nuclear ribonucleoprotein M (hnRNP M) as a key molecule controlling D2R splicing. We show that binding of hnRNP M to exon 6 inhibited the inclusion of this exon in the mRNA. Importantly, the splicing factor Nova-1 counteracted hnRNP M effects on D2R pre-mRNA splicing. Indeed, mutations of the putative Nova-1-binding site on exon 6 disrupted Nova-1 RNA assembly and diminished the inhibitory effect of Nova-1 on hnRNP M-dependent exon 6 exclusion. These results identify Nova-1 and hnRNP M as D2R pre-mRNA-binding proteins and show their antagonistic role in the alternative splicing of D2R pre-mRNA. PMID- 21622565 TI - Presenilin regulates insulin signaling via a gamma-secretase-independent mechanism. AB - Presenilin (PS), a causative molecule of familial Alzheimer disease, acts as a crucial component of the gamma-secretase complex, which is required to cleave type I transmembrane proteins such as amyloid precursor protein and Notch. However, it also functions through gamma-secretase-independent pathways. Recent reports suggested that PS could regulate the expression level of cell surface receptors, including the PDGF and EGF receptors, followed by modulating their downstream pathways via gamma-secretase-independent mechanisms. The main purpose of this study was to clarify the effect of PS on expression of the insulin receptor (IR) as well as on insulin signaling. Here, we demonstrate that PS inhibited IR transcription and reduced IR expression, and this was followed by down-regulation of insulin signaling. Moreover, we suggest that neither gamma secretase activity nor Wnt/beta-catenin signaling can reduce the expression of IR, but a PS-mediated increase in the intracellular Ca(2+) level can be associated with it. These results clearly indicate that PS can functionally regulate insulin signaling by controlling IR expression. PMID- 21622566 TI - Pleiotropic and age-dependent effects of decreased protein modification by O linked N-acetylglucosamine on pancreatic beta-cell function and vascularization. AB - The hexosamine biosynthesis pathway (HBP) regulates the post-translational modification of nuclear and cytoplasmic protein by O-linked N-acetylglucosamine (O-GlcNAc). Numerous studies have demonstrated increased flux through this pathway contributes to the development of beta-cell dysfunction. The effect of decreased O-GlcNAc on the maintenance of normal beta-cell function, however, is not well understood. We studied transgenic mice that over express beta-N acetylglucosaminidase (O-GlcNAcase), an enzyme that catalyzes the removal of O GlcNAc from proteins, in the pancreatic beta-cell under control of the rat insulin promoter. 3-4-Month-old O-GlcNAcase transgenic mice have higher glucose excursions with a concomitant decrease in circulating insulin levels, insulin mRNA levels, and total islet insulin content. In older (8-9-month-old) O GlcNAcase transgenic mice glucose tolerance is no longer impaired. This is associated with increased serum insulin, islet insulin content, and insulin mRNA in the O-GlcNAcase transgenic mice. These improvements in beta-cell function with aging are associated with increased angiogenesis and increased VEGF expression, with parallel increases in activation of Akt and expression of PGC1alpha. The biphasic effects as a function of age are consistent with published observations of mice with increased O-GlcNAc in islets and demonstrate that O-GlcNAc signaling exerts multiple effects on both insulin secretion and islet survival. PMID- 21622567 TI - Analyses of fruit flies that do not express selenoproteins or express the mouse selenoprotein, methionine sulfoxide reductase B1, reveal a role of selenoproteins in stress resistance. AB - Selenoproteins are essential in vertebrates because of their crucial role in cellular redox homeostasis, but some invertebrates that lack selenoproteins have recently been identified. Genetic disruption of selenoprotein biosynthesis had no effect on lifespan and oxidative stress resistance of Drosophila melanogaster. In the current study, fruit flies with knock-out of the selenocysteine-specific elongation factor were metabolically labeled with (75)Se; they did not incorporate selenium into proteins and had the same lifespan on a chemically defined diet with or without selenium supplementation. These flies were, however, more susceptible to starvation than controls, and this effect could be ascribed to the function of selenoprotein K. We further expressed mouse methionine sulfoxide reductase B1 (MsrB1), a selenoenzyme that catalyzes the reduction of oxidized methionine residues and has protein repair function, in the whole body or the nervous system of fruit flies. This exogenous selenoprotein could only be expressed when the Drosophila selenocysteine insertion sequence element was used, whereas the corresponding mouse element did not support selenoprotein synthesis. Ectopic expression of MsrB1 in the nervous system led to an increase in the resistance against oxidative stress and starvation, but did not affect lifespan and reproduction, whereas ubiquitous MsrB1 expression had no effect. Dietary selenium did not influence lifespan of MsrB1-expressing flies. Thus, in contrast to vertebrates, fruit flies preserve only three selenoproteins, which are not essential and play a role only under certain stress conditions, thereby limiting the use of the micronutrient selenium by these organisms. PMID- 21622568 TI - Mitochondrial carnitine palmitoyltransferase 1a (CPT1a) is part of an outer membrane fatty acid transfer complex. AB - CPT1a (carnitine palmitoyltransferase 1a) in the liver mitochondrial outer membrane (MOM) catalyzes the primary regulated step in overall mitochondrial fatty acid oxidation. It has been suggested that the fundamental unit of CPT1a exists as a trimer, which, under native conditions, could form a dimer of the trimers, creating a hexamer channel for acylcarnitine translocation. To examine the state of CPT1a in the MOM, we employed a combined approach of sizing by mass and isolation using an immunological method. Blue native electrophoresis followed by detection with immunoblotting and mass spectrometry identified large molecular mass complexes that contained not only CPT1a but also long chain acyl-CoA synthetase (ACSL) and the voltage-dependent anion channel (VDAC). Immunoprecipitation with antisera against the proteins revealed a strong interaction between the three proteins. Immobilized CPT1a-specific antibodies immunocaptured not only CPT1a but also ACSL and VDAC, further strengthening findings with blue native electrophoresis and immunoprecipitation. This study shows strong protein-protein interaction between CPT1a, ACSL, and VDAC. We propose that this complex transfers activated fatty acids through the MOM. PMID- 21622569 TI - ICP34.5 protein of herpes simplex virus facilitates the initiation of protein translation by bridging eukaryotic initiation factor 2alpha (eIF2alpha) and protein phosphatase 1. AB - The ICP34.5 protein of herpes simplex virus type 1 is a neurovirulence factor that plays critical roles in viral replication and anti-host responses. One of its functions is to recruit protein phosphatase 1 (PP1) that leads to the dephosphorylation of the alpha subunit of translation initiation factor eIF2 (eIF2alpha), which is inactivated by infection-induced phosphorylation. As PP1 is a protein phosphatase with a wide range of substrates, the question remains to be answered how ICP34.5 directs PP1 to specifically dephosphorylate eIF2alpha. Here we report that ICP34.5 not only binds PP1 but also associates with eIF2alpha by in vitro and in vivo assays. The binding site of eIF2alpha is identified at amino acids 233-248 of ICP34.5, which falls in the highly homologous region with human gene growth arrest and DNA damage 34. The interaction between ICP34.5 and eIF2alpha is independent of the phosphorylation status of eIF2alpha at serine 51. Deletion mutation of this region results in the failure of dephosphorylation of eIF2alpha by PP1 and, consequently, interrupts viral protein synthesis and replication. Our data illustrated that the binding between viral protein ICP34.5 and the host eIF2alpha is crucial for the specific dephosphorylation of eIF2alpha by PP1. We propose that herpes simplex virus protein ICP34.5 bridges PP1 and eIF2alpha via their binding motifs and thereby facilitates the protein synthesis and viral replication. PMID- 21622570 TI - Forkhead-associated (FHA) domain containing ABC transporter Rv1747 is positively regulated by Ser/Thr phosphorylation in Mycobacterium tuberculosis. AB - One major signaling method employed by Mycobacterium tuberculosis, the causative agent of tuberculosis, is through reversible phosphorylation of proteins mediated by protein kinases and phosphatases. This study concerns one of these enzymes, the serine/threonine protein kinase PknF, that is encoded in an operon with Rv1747, an ABC transporter that is necessary for growth of M. tuberculosis in vivo and contains two forkhead-associated (FHA) domains. FHA domains are phosphopeptide recognition motifs that specifically recognize phosphothreonine containing epitopes. Experiments to determine how PknF regulates the function of Rv1747 demonstrated that phosphorylation occurs on two specific threonine residues, Thr-150 and Thr-208. To determine the in vivo consequences of phosphorylation, infection experiments were performed in bone marrow-derived macrophages and in mice using threonine-to-alanine mutants of Rv1747 that prevent specific phosphorylation and revealed that phosphorylation positively modulates Rv1747 function in vivo. The role of the FHA domains in this regulation was further demonstrated by isothermal titration calorimetry, using peptides containing both phosphothreonine residues. FHA-1 domain mutation resulted in attenuation in macrophages highlighting the critical role of this domain in Rv1747 function. A mutant deleted for pknF did not, however, have a growth phenotype in an infection, suggesting that other kinases can fulfill its role when it is absent. This study provides the first information on the molecular mechanism(s) regulating Rv1747 through PknF-dependent phosphorylation but also indicates that phosphorylation activates Rv1747, which may have important consequences in regulating growth of M. tuberculosis. PMID- 21622571 TI - NF-kappaB essential modulator (NEMO) interaction with linear and lys-63 ubiquitin chains contributes to NF-kappaB activation. AB - The IkappaB kinase (IKK) complex acts as a gatekeeper of canonical NF-kappaB signaling in response to upstream stimulation. IKK activation requires sensing of ubiquitin chains by the essential IKK regulatory subunit IKKgamma/NEMO. However, it has remained enigmatic whether NEMO binding to Lys-63-linked or linear ubiquitin chains is critical for triggering IKK activation. We show here that the NEMO C terminus, comprising the ubiquitin binding region and a zinc finger, has a high preference for binding to linear ubiquitin chains. However, immobilization of NEMO, which may be reminiscent of cellular oligomerization, facilitates the interaction with Lys-63 ubiquitin chains. Moreover, selective mutations in NEMO that abolish association with linear ubiquitin but do not affect binding to Lys 63 ubiquitin are only partially compromising NF-kappaB signaling in response to TNFalpha stimulation in fibroblasts and T cells. In line with this, TNFalpha triggered expression of NF-kappaB target genes and induction of apoptosis was partially compromised by NEMO mutations that selectively impair the binding to linear ubiquitin chains. Thus, in vivo NEMO interaction with linear and Lys-63 ubiquitin chains is required for optimal IKK activation, suggesting that both type of chains are cooperating in triggering canonical NF-kappaB signaling. PMID- 21622572 TI - Engineered bivalent ligands to bias ErbB receptor-mediated signaling and phenotypes. AB - The ErbB receptor family is dysregulated in many cancers, and its therapeutic manipulation by targeted antibodies and kinase inhibitors has resulted in effective chemotherapies. However, many malignancies remain refractory to current interventions. We describe a new approach that directs ErbB receptor interactions, resulting in biased signaling and phenotypes. Due to known receptor ligand affinities and the necessity of ErbB receptors to dimerize to signal, bivalent ligands, formed by the synthetic linkage of two neuregulin-1beta (NRG) moieties, two epidermal growth factor (EGF) moieties, or an EGF and a NRG moiety, can potentially drive homotypic receptor interactions and diminish formation of HER2-containing heterodimers, which are implicated in many malignancies and are a prevalent outcome of stimulation by native, monovalent EGF, or NRG. We demonstrate the therapeutic potential of this approach by showing that bivalent NRG (NN) can bias signaling in HER3-expressing cancer cells, resulting in some cases in decreased migration, inhibited proliferation, and increased apoptosis, whereas native NRG stimulation increased the malignant potential of the same cells. Hence, this new approach may have therapeutic relevance in ovarian, breast, lung, and other cancers in which HER3 has been implicated. PMID- 21622573 TI - Dimer interface of the effector domain of non-structural protein 1 from influenza A virus: an interface with multiple functions. AB - Non-structural protein 1 from influenza A virus, NS1A, is a key multifunctional virulence factor composed of two domains: an N-terminal double-stranded RNA (dsRNA)-binding domain and a C-terminal effector domain (ED). Isolated RNA binding and effector domains of NS1A both exist as homodimers in solution. Despite recent crystal structures of isolated ED and full-length NS1A proteins from different influenza virus strains, controversy remains over the actual biologically relevant ED dimer interface. Here, we report the biophysical properties of the NS1A ED from H3N2 influenza A/Udorn/307/1972 (Ud) virus in solution. Several lines of evidence, including (15)N NMR relaxation, NMR chemical shift perturbations, static light scattering, and analytical sedimentation equilibrium, demonstrate that Ud NS1A ED forms a relatively weak dimer in solution (K(d) = 90 +/- 2 MUm), featuring a symmetric helix-helix dimer interface. Mutations within and near this interface completely abolish dimerization, whereas mutations consistent with other proposed ED dimer interfaces have no effect on dimer formation. In addition, the critical Trp-187 residue in this interface serves as a sensitive NMR spectroscopic marker for the concentration-dependent dimerization of NS1A ED in solution. Finally, dynamic light scattering and gel shift binding experiments demonstrate that the ED interface plays a role in both the oligomerization and the dsRNA binding properties of the full-length NS1A protein. In particular, mutation of the critical tryptophan in the ED interface substantially reduces the propensity of full-length NS1A from different strains to oligomerize and results in a reduction in dsRNA binding affinity for full-length NS1A. PMID- 21622574 TI - Skeletal muscle differentiation and fusion are regulated by the BAR-containing Rho-GTPase-activating protein (Rho-GAP), GRAF1. AB - Although RhoA activity is necessary for promoting myogenic mesenchymal stem cell fates, recent studies in cultured cells suggest that down-regulation of RhoA activity in specified myoblasts is required for subsequent differentiation and myotube formation. However, whether this phenomenon occurs in vivo and which Rho modifiers control these later events remain unclear. We found that expression of the Rho-GTPase-activating protein, GRAF1, was transiently up-regulated during myogenesis, and studies in C2C12 cells revealed that GRAF1 is necessary and sufficient for mediating RhoA down-regulation and inducing muscle differentiation. Moreover, forced expression of GRAF1 in pre-differentiated myoblasts drives robust muscle fusion by a process that requires GTPase activating protein-dependent actin remodeling and BAR-dependent membrane binding or sculpting. Moreover, morpholino-based knockdown studies in Xenopus laevis determined that GRAF1 expression is critical for muscle development. GRAF1 depleted embryos exhibited elevated RhoA activity and defective myofibrillogenesis that resulted in progressive muscle degeneration, defective motility, and embryonic lethality. Our results are the first to identify a GTPase activating protein that regulates muscle maturation and to highlight the functional importance of BAR domains in myotube formation. PMID- 21622575 TI - Molecular mechanism of the E99K mutation in cardiac actin (ACTC Gene) that causes apical hypertrophy in man and mouse. AB - We generated a transgenic mouse model expressing the apical hypertrophic cardiomyopathy-causing mutation ACTC E99K at 50% of total heart actin and compared it with actin from patients carrying the same mutation. The actin mutation caused a higher Ca(2+) sensitivity in reconstituted thin filaments measured by in vitro motility assay (2.3-fold for mice and 1.3-fold for humans) and in skinned papillary muscle. The mutation also abolished the change in Ca(2+) sensitivity normally linked to troponin I phosphorylation. MyBP-C and troponin I phosphorylation levels were the same as controls in transgenic mice and human carrier heart samples. ACTC E99K mice exhibited a high death rate between 28 and 45 days (48% females and 22% males). At 21 weeks, the hearts of the male survivors had enlarged atria, increased interstitial fibrosis, and sarcomere disarray. MRI showed hypertrophy, predominantly at the apex of the heart. End diastolic volume and end-diastolic pressure were increased, and relaxation rates were reduced compared with nontransgenic littermates. End-systolic pressures and volumes were unaltered. ECG abnormalities were present, and the contractile response to beta-adrenergic stimulation was much reduced. Older mice (29-week-old females and 38-week-old males) developed dilated cardiomyopathy with increased end-systolic volume and continuing increased end-diastolic pressure and slower contraction and relaxation rates. ECG showed atrial flutter and frequent atrial ectopic beats at rest in some ACTC E99K mice. We propose that the ACTC E99K mutation causes higher myofibrillar Ca(2+) sensitivity that is responsible for the sudden cardiac death, apical hypertrophy, and subsequent development of heart failure in humans and mice. PMID- 21622576 TI - The Ets factor Etv1 interacts with Tpit protein for pituitary pro opiomelanocortin (POMC) gene transcription. AB - Pro-opiomelanocortin (POMC) is expressed in two lineages of the pituitary, the anterior lobe corticotrophs and the intermediate lobe melanotrophs. POMC expression in these two lineages is highly dependent on the cell-restricted transcription factor Tpit. As Tpit intervenes relatively late in differentiation of those lineages, we have been searching for other transcription factors that may participate in their gene expression program. On the basis of similarity with the Tpit expression profile, we identified Ets variant gene 1 (Etv1/Er81) as a putative POMC transcription factor. Using Etv1-lacZ knockin mice, we describe preferential Etv1 expression in pituitary POMC cells and also in posterior lobe pituicytes. We further show that Etv1 enhances POMC transcription on its own and in synergy with Tpit. The Ets-binding site located within the Tpit/Pitx regulatory element is necessary for Etv1 activity in POMC-expressing AtT-20 cells but dispensable for synergy with Tpit. Etv1 and Tpit interact together in coimmunoprecipitation experiments. Furthermore, Etv1 is present at the POMC promoter, and siRNA-mediated knockdown of Etv1 in AtT-20 cells produces a significant decrease in POMC expression. Etv1 knockout pituitaries show normal POMC cell distribution and normal POMC mRNA abundance, suggesting compensation by other factors. The coordinate expression of Etv1 with POMC cell differentiation and its interaction with the highly cell-restricted Tpit factor indicate that Etv1 participates in a combinatorial code for pituitary cell-specific gene expression. PMID- 21622577 TI - The opening of the SPP1 bacteriophage tail, a prevalent mechanism in Gram positive-infecting siphophages. AB - The SPP1 siphophage uses its long non-contractile tail and tail tip to recognize and infect the Gram-positive bacterium Bacillus subtilis. The tail-end cap and its attached tip are the critical components for host recognition and opening of the tail tube for genome exit. In the present work, we determined the cryo electron microscopic (cryo-EM) structure of a complex formed by the cap protein gp19.1 (Dit) and the N terminus of the downstream protein of gp19.1 in the SPP1 genome, gp21(1-552) (Tal). This complex assembles two back-to-back stacked gp19.1 ring hexamers, interacting loosely, and two gp21(1-552) trimers interacting with gp19.1 at both ends of the stack. Remarkably, one gp21(1-552) trimer displays a "closed" conformation, whereas the second is "open" delineating a central channel. The two conformational states dock nicely into the EM map of the SPP1 cap domain, respectively, before and after DNA release. Moreover, the open/closed conformations of gp19.1-gp21(1-552) are consistent with the structures of the corresponding proteins in the siphophage p2 baseplate, where the Tal protein (ORF16) attached to the ring of Dit (ORF15) was also found to adopt these two conformations. Therefore, the present contribution allowed us to revisit the SPP1 tail distal-end architectural organization. Considering the sequence conservation among Dit and the N-terminal region of Tal-like proteins in Gram-positive infecting Siphoviridae, it also reveals the Tal opening mechanism as a hallmark of siphophages probably involved in the generation of the firing signal initiating the cascade of events that lead to phage DNA release in vivo. PMID- 21622578 TI - Trends of inpatient spine augmentation: 2001-2008. AB - BACKGROUND AND PURPOSE: Vertebroplasty and kyphoplasty are 2 minimally invasive spine augmentation procedures currently used in the management of vertebral compression fractures. Our aim was to examine the NIS data base to identify trends in spine augmentation procedures over time. MATERIALS AND METHODS: Patients were identified from the NIS data base by using primary diagnosis codes (ICD-9-pathologic vertebral fracture, 733.13) and procedures codes (ICD-9-other repair of bone, 78.49; vertebroplasty, 81.65; kyphoplasty, 81.66). Means and their corresponding standard errors were compared for statistically significant differences by using the Z-test. RESULTS: Between 2001 and 2008, >240 000 inpatient spine augmentations were performed in the United States. From 2001 to 2008, there was a 741% increase in the number of hospital discharges for patients who underwent a spine augmentation, but the year-to-year rate of increase has been declining since 2001. From 2004 to 2008, >50 000 inpatient verterbroplasties and >152 000 inpatient kyphoplasties were identified. Compared with vertebroplasty, kyphoplasty patients were less often admitted from the emergency department (33% versus 56%, P < .001), had shorter postaugmentation hospital stays (2.3 versus 3.1 days, P < .001), had fewer comorbidities at presentation (1.4 versus 1.6, P < .01), and were less likely to be discharged to a long-term facility (28% versus 43%, P < .001). CONCLUSIONS: Significant differences were noted in multiple comparisons between vertebroplasty and kyphoplasty, including length of hospital stay and discharges to long-term facilities. However, these findings may simply reflect differences in practice patterns rather than real differences in efficacy between the procedures. PMID- 21622579 TI - Computational modeling and flow diverters: a teaching moment. PMID- 21622580 TI - Intraoperative angiography for cranial dural arteriovenous fistula. AB - BACKGROUND AND PURPOSE: IA is a valuable adjunct during surgery for a variety of neurovascular diseases; however, there are no reported series describing IA for DAVFs. This study was undertaken to evaluate the safety and efficacy of IA for DAVFs. MATERIALS AND METHODS: A retrospective review of DAVF surgical cases during a 20-year period was conducted, and cases with IA were evaluated. Clinical details, surgical and angiographic findings, and postoperative outcomes were reviewed. The incidence of residual fistula on IAs, the utility of the surgical procedure, and the incidence of false-negative findings on IA were also determined. RESULTS: IA was performed in 29 patients (31 DAVFs) for DAVFs. The distribution of the fistulas was the following: transverse-sigmoid (n = 9), tentorial (n = 6), torcular (n = 3), cavernous sinus (n = 4), SSS (n = 4), foramen magnum (n = 3), and temporal-middle fossa (n = 2). Twelve patients had undergone prior embolization, while 6 patients had unsuccessful embolization procedures. Thirty-eight surgeries were performed for DAVF in 29 patients, and IA was performed in 34 surgeries. Forty-four angiographic procedures were performed in the 34 surgeries. Nine patients underwent multiple angiographies. In 11 patients (37.9%), IA revealed residual fistula after the surgeon determined that no lesion remained. This led to further exploration at the same sitting in 10 patients, while in 1 patient, further surgery was performed at a later date. False-negative findings on IA occurred in 3 patients (10.7%). CONCLUSIONS: IA is an important adjunct in surgery for DAVF. In this series, it resulted in further surgical treatment in 37.9% of patients. However, there was a 10% false-negative rate, which justified subsequent postoperative angiography. PMID- 21622581 TI - Nonbifurcating cervical carotid artery diagnosed by MR angiography. AB - BACKGROUND AND PURPOSE: A nonbifurcating cervical carotid artery is a branching anomaly in which the ECA has no proximal main trunk. We report its incidence and characteristic features on MRA. MATERIALS AND METHODS: We retrospectively reviewed MRAs of 2866 patients obtained by using a standard noncontrast MRA protocol and two 1.5T MR imaging units and reviewed the English language literature to assess the occurrence and features of this nonbifurcating artery. RESULTS: We diagnosed 6 cases, indicating an incidence of 0.21%, and found 11 cases reported in the literature. Analysis of all 17 cases demonstrated no laterality or sex predominance. The most prevalent pattern of branching order from proximal to distal was the F-L trunk, the distal trunk of the ECA, and the OA. CONCLUSIONS: A nonbifurcating cervical carotid artery is rare but not as extremely rare as previously considered, and its correct diagnosis is necessary to avoid complications during interventional radiologic procedures or head and neck surgeries. PMID- 21622582 TI - The TERT-CLPTM1L locus for lung cancer predisposes to bronchial obstruction and emphysema. AB - Clinical studies suggest that bronchial obstruction and emphysema increase susceptibility to lung cancer. We assessed the possibility of a common genetic origin and investigated whether the lung cancer susceptibility locus on chromosome 5p15.33 increases the risk for bronchial obstruction and emphysema. Three variants in the 5p15.33 locus encompassing the TERT and CLPTM1L genes were genotyped in 777 heavy smokers and 212 lung cancer patients. Participants underwent pulmonary function tests and computed tomography of the chest, and completed questionnaires assessing smoking behaviour. The rs31489 C-allele correlated with reduced forced expiratory volume in 1 s (p=0.006). Homozygous carriers of the rs31489 C-allele exhibited increased susceptibility to bronchial obstruction (OR 1.82, 95% CI 1.24-2.69; p=0.002). A similar association was observed for diffusing capacity of the lung for carbon monoxide (p=0.004). Consistent with this, CC-carriers had an increased risk of emphysema (OR 2.04, 95% CI 1.41-2.94; p=1.73 * 10(-4)) and displayed greater alveolar destruction. Finally, CC-carriers also had an increased risk for lung cancer (OR 1.90, 95% CI 1.21-2.99; p=0.005), and were more susceptible to developing both lung cancer and bronchial obstruction than lung cancer alone (OR 2.11, 95% CI 1.04-4.26; p=0.038). The rs31489 variant on 5p15.33 is associated with bronchial obstruction, presence and severity of emphysema, and lung cancer. PMID- 21622583 TI - The European Sleep Apnoea Database (ESADA): report from 22 European sleep laboratories. AB - The European Sleep Apnoea Database (ESADA) reflects a network of 22 sleep disorder centres in Europe enabled by a COST action B26 programme. This ongoing project aims to describe differences in standard clinical care of patients with obstructive sleep apnoea (OSA) and to establish a resource for genetic research in this disorder. Patients with suspected OSA are consecutively included and followed up according to local clinical standards. Anthropometrics, medical history, medication, daytime symptoms and sleep data (polysomnography or cardiorespiratory polygraphy) are recorded in a structured web-based report form. 5,103 patients (1,426 females, mean+/-sd age 51.8+/-12.6 yrs, 79.4% with apnoea/hypopnoea index (AHI) >=5 events.h(-1)) were included from March 15, 2007 to August 1, 2009. Morbid obesity (body mass index >=35 kg.m(-2)) was present in 21.1% of males and 28.6% of females. Cardiovascular, metabolic and pulmonary comorbidities were frequent (49.1%, 32.9% and 14.2%, respectively). Patients investigated with a polygraphic method had a lower AHI than those undergoing polysomnography (23.2+/-23.5 versus 29.1+/-26.3 events.h(-1), p<0.0001). The ESADA is a rapidly growing multicentre patient cohort that enables unique outcome research opportunities and genotyping. The first cross-sectional analysis reveals a high prevalence of cardiovascular and metabolic morbidity in patients investigated for OSA. PMID- 21622584 TI - Reference values of exhaled nitric oxide in healthy Asian children aged 5 to 18 years. AB - This study was undertaken to establish reference values of exhaled nitric oxide fraction (F(eNO)) and its determinants in healthy Asian children. 693 healthy Asian children aged 5-18 yrs were assessed using a single-breath online F(eNO) measurement (exhaled flow 50 mL.s(-1)), questionnaires, anthropometric measurements, spirometry and total and specific immunoglobulin (Ig) E. Geometric mean F(eNO) and the upper 95% CI were 13.7 ppb and 49.7 ppb, respectively, for healthy children, and 11.2 ppb and 30.2 ppb, respectively, for those without allergic sensitisation. F(eNO) was positively associated with age, allergic sensitisation, total IgE, ambient nitric oxide, measurement in the afternoon, and drinking water within 1 h before testing, and was negatively associated with weight. In healthy children without allergic sensitisation, age was the single best explanatory variable. The F(eNO) predicted values were 1-2 ppb higher in Asian than in Caucasian children in earlier studies, while the upper 95% CI were 9-10 ppb higher. In conclusion, the upper limits of normal F(eNO) in Asian children depend on age, from 21 ppb in young children to 39 ppb in adolescents. Ethnicity, age, allergic sensitisation, total IgE, ambient nitric oxide, time of testing, drinking water and weight are important determinants. PMID- 21622585 TI - Validation and comparison of several published prognostic systems for patients with small cell lung cancer. AB - The aim of the present study was to validate and compare published prognostic classifications for predicting the survival of patients with small cell lung cancer. We pooled data from phase III randomised clinical trials, and used Cox models for validation purposes and concordance probability estimates for assessing predictive ability. We included 693 patients. All the classifications impacted significantly on survival, with hazard ratios (HRs) in the range 1.57 1.68 (all p<0.0001). Median survival times were 16-19 months for the best predicted groups, while they were 6-7 months for the most poorly predicted groups. Most of the paired comparisons were statistically significant. We obtained similar results when restricting the analysis to patients with extensive disease. Multivariate Cox models for fitting survival data were also performed. The HRs for a single covariate were 8.23 (95% CI 5.88-11.69), and 9.46 (6.67 13.50), and for extensive disease were 5.60 (3.13-9.93), 12.49 (5.57-28.01) and 8.83 (4.66-16.64). Concordance probability estimates ranged 0.55-0.65 (overlapping confidence intervals). Published classifications were validated and suitable for use at a population level. As expected, prediction at an individual level remains problematic. A specific model designed for extensive-disease patients did not appear to perform better. PMID- 21622586 TI - Computed tomography measurements of parapneumonic effusion indicative of thoracentesis. AB - Patients with parapneumonic effusions (PPE) measuring <1 cm by lateral decubitus radiograph (LDR) or <5 cm by lateral erect radiograph (LER) do not require thoracentesis. No such data exist for chest computed tomography (CCT). The objective of this study was to identify a PPE measurement by CCT that indicates the need for thoracentesis. A secondary data analysis of two pneumonia databases was conducted to identify patients with PPE. Measurements of PPE using LDR, LER and CCT were correlated by linear regression analysis. The clinical outcome of community-acquired pneumonia patients managed with the newly defined CCT measurement was evaluated. PPE was identified in 419 out of 1,460 patients with possible pneumonia. PPE measurements of 1 cm and 5 cm by LDR and LER, respectively, correlated with a measurement of 2.5 cm by CCT. Out of 95 patients with CCT measurements <2.5 cm, 31 poor clinical outcomes were reported: outcome was PPE related (n = 1); outcome was PPE unrelated (n = 26); and outcome was not evaluable (n = 4). The single case of poor outcome also measured <1 cm by LDR. This study indicates that patients with community-acquired pneumonia and a PPE measuring <2.5 cm by CCT can be managed without the need for thoracentesis. PMID- 21622587 TI - Blinded 12-week comparison of once-daily indacaterol and tiotropium in COPD. AB - Two, once daily (q.d.) inhaled bronchodilators are available for the treatment of chronic obstructive pulmonary disease (COPD): the beta(2)-agonist indacaterol and the anticholinergic tiotropium. This blinded study compared the efficacy of these two agents and assessed their safety and tolerability. Patients with moderate-to severe COPD were randomised to treatment with indacaterol 150 MUg q.d. (n=797) or tiotropium 18 MUg q.d. (n=801) for 12 weeks. After 12 weeks, the two treatments had similar overall effects on "trough" (24 h post-dose) forced expiratory volume in 1 s. Indacaterol-treated patients had greater improvements in transition dyspnoea index (TDI) total score (least squares means 2.01 versus 1.43; p<0.001) and St George's Respiratory Questionnaire (SGRQ) total score (least squares means 37.1 versus 39.2; p<0.001; raw mean change from baseline -5.1 versus -3.0), and were significantly more likely to achieve clinically relevant improvements in these end-points (indacaterol versus tiotropium odds ratios of 1.49 for TDI and 1.43 for SGRQ, both p<0.001). Adverse events were recorded for 39.7% and 37.2% of patients in the indacaterol and tiotropium treatment groups, respectively. The most frequent adverse events were COPD worsening, cough and nasopharyngitis. Both bronchodilators demonstrated spirometric efficacy. The two treatments were well tolerated with similar adverse event profiles. Compared with tiotropium, indacaterol provided significantly greater improvements in clinical outcomes. PMID- 21622588 TI - Role of IL-1alpha and the Nlrp3/caspase-1/IL-1beta axis in cigarette smoke induced pulmonary inflammation and COPD. AB - Cigarette smoke (CS), the primary risk factor of chronic obstructive pulmonary disease (COPD), leads to pulmonary inflammation through interleukin-1 receptor (IL-1R)I signalling, as determined using COPD mouse models. It is unclear whether interleukin (IL)-1alpha or IL-1beta, activated by the Nlrp3/caspase-1 axis, is the predominant ligand for IL-1RI in CS-induced responses. We exposed wild-type mice (treated with anti-IL-1alpha or anti-IL-1beta antibodies), and IL-1RI knockout (KO), Nlrp3 KO and caspase-1 KO mice to CS for 3 days or 4 weeks and evaluated pulmonary inflammation. Additionally, we measured the levels of IL 1alpha and IL-1beta mRNA (in total lung tissue by RT-PCR) and protein (in induced sputum by ELISA) of never-smokers, smokers without COPD and patients with COPD. In CS-exposed mice, pulmonary inflammation was dependent on IL-1RI and could be significantly attenuated by neutralising IL-1alpha or IL-1beta. Interestingly, CS induced inflammation occurred independently of IL-1beta activation by the Nlrp3/caspase-1 axis. In human subjects, IL-1alpha and IL-1beta were significantly increased in total lung tissue and induced sputum of patients with COPD, respectively, compared with never-smokers. These results suggest that not only IL-1beta but also IL-1alpha should be considered as an important mediator in CS-induced inflammation and COPD. PMID- 21622589 TI - Do responses to exercise training in cystic fibrosis depend on initial fitness level? AB - The aim of our study was to evaluate the responses to an exercise programme with respect to initial fitness in subjects with cystic fibrosis (CF). 72 subjects (42 female) aged 10-43 yrs (forced expiratory volume in 1 s of 62.0 +/- 26.7% predicted) were included. Participants were divided into three groups based on peak oxygen uptake expressed as % predicted. Subjects participated in a multifaceted rehabilitation programme, including five-times-a-week exercise training, for 6 weeks. Exercise and ventilatory capacity were determined by a maximal incremental cycling test. Oxygen uptake, workload and peak cardiac frequency at peak and submaximal workload were used as parameters for exercise capacity and responsiveness to training. Lung function values were significantly different between groups (p < 0.05), and increased after training (p < 0.05) only in groups with a lower initial fitness level. Responsiveness to training showed differences between groups (p < 0.05) at peak and ventilatory anaerobic threshold (VAT; p < 0.05), with higher improvements in subjects with lower initial fitness level. The improvements in exercise parameters in CF at peak and VAT depended on the fitness level at baseline, independent of lung function. These improvements seen after training were comparable with those seen in healthy subjects, suggesting that responsiveness to exercise is similar in CF and in healthy untrained persons. PMID- 21622590 TI - Obstructive sleep apnoea and metabolic impairment in severe obesity. AB - Obstructive sleep apnoea (OSA) seems to worsen metabolism. This effect has not been evaluated in morbid obesity (MO). We hypothesised that the metabolic profile is more impaired in MO patients with OSA than in those without, and investigated whether any specific metabolic dysfunction is related to OSA in MO. A prospective multicentre cross-sectional study was conducted in consecutive subjects before bariatric surgery. OSA was defined as apnoea/hypopnoea index (AHI) >=15 by overnight polysomnography. Anthropometrical, blood pressure (BP) and fasting blood measurements were obtained the morning after. Metabolic syndrome (MetS) was defined according to National Cholesterol Education Program Adult Treatment Panel III modified criteria. 159 patients were studied: 72% were female and 72% had OSA. MetS prevalence was 70% in OSA versus 36% in non-OSA (p<0.001). As AHI severity increased, metabolic parameters progressively worsened, even in those without type 2 diabetes (DM2). AHI was independently associated with systolic and diastolic BP, triglycerides and the percentage of glycosylated haemoglobin (HbA1c) in the total sample, and with systolic BP, high-density lipoprotein cholesterol and HbA1c in those samples without DM2. OSA increased the adjusted odds ratio of having MetS by 2.8 (95% CI 1.3-6.2; p=0.009). In MO, OSA is associated with major metabolic impairment caused by higher BP and poorer lipid and glucose control, independent of central obesity or DM2. PMID- 21622591 TI - Self-reported sleep apnoea and mortality in patients from the Swedish Obese Subjects study. AB - Sleep apnoea is associated with increased mortality in sleep clinic and community population groups. It is unclear whether a clinical report of sleep apnoea results in additional mortality risk in patients with severe obesity. The Swedish Obese Subjects (SOS) study is a nonrandomised controlled trial of bariatric surgery versus conventional treatment for the treatment of severe obesity and its complications (mean +/- SD body mass index 41 +/- 5 kg . m(-2)). The presence or absence of sleep apnoea (witnessed pauses in breathing) was determined by self reporting at baseline in 3,953 patients who were observed for 54,236 person-yrs (mean 13.5 maximum 21.0 yrs). Sleep apnoea was reported by 934 (23.6%) patients at baseline and was a significant univariate predictor of mortality (hazard ratio (95% CI) 1.74 (1.40-2.18)). In a range of multivariate models of mortality risk, controlling for <= 16 other potential confounders and established mortality risk factors, sleep apnoea remained a significant prognostic factor (fully adjusted model 1.29 (1.01-1.65)). Self-reported sleep apnoea is an independent prognostic marker of all-cause mortality in obese patients. PMID- 21622592 TI - The relationship between relapse, impairment and disability in multiple sclerosis. AB - OBJECTIVE: To describe the spatial relationship between relapse and disability in multiple sclerosis (MS). METHODS: 141 relapse onset MS patients were studied. For each patient an examination was performed and a relapse history obtained. Multivariate logistic regression examined whether there was an association between localizing clinical signs and a history of relevant relapse in order to explore the spatial relationship between relapse and subsequent disability. RESULTS: The presence of impaired vision or sensation was independently associated with a history of one or more anatomically related relapses. The presence of weakness or cerebellar ataxia in a limb was not associated with a single relevant relapse but was associated with multiple relevant relapses. A history of multiple episodes of weakness or ataxia in the same limb was uncommon. CONCLUSIONS: Our data suggest that motor pathways are relatively resistant to chronic impairment from acute relapse, whereas afferent pathways are more susceptible. This, in combination with prominent usage of the Expanded Disability Status Scale, which is dependent on mobility and motor function at higher scores, may explain the paradox between natural history studies that suggest relapses are irrelevant to long-term disability and shorter studies at lower disability levels suggesting relapses are responsible for disability accumulation. PMID- 21622593 TI - Community walking can be assessed using a 10-metre timed walk test. AB - BACKGROUND: A decline in mobility is a common feature of multiple sclerosis (MS). Community walking scales are used to categorize patients in their ability to move independently. The first purpose of this study was to determine which specific gait speed corresponded with the categories of the Modified Functional Walking Categories (MFWC). The second purpose was to determine the Minimally Important Change (MIC) in absolute gait speed using the MFWC and Expanded Disability Status Scale (EDSS) as external criteria. METHOD: MS patients were measured six times in 6 years. Gait velocity was measured with the 10-metre timed walk test (10-m TWT), the severity of MS was determined with the EDSS, and community walking was assessed with the MFWC. For each category of the MFWC, Receiver Operating Characteristic (ROC) curves were used to find the best possible cut-off point on the 10-m TWT. The MIC in absolute gait speed was determined using a change of one category on the MFWC or one point on the EDSS. RESULTS: A strong relationship was found between gait speed and the MFWC; all areas under the ROC curves (AUCs) were between 0.74 and 0.86. The MIC in absolute gait speed could not be determined, because the AUCs were below the threshold of 0.70 and changes in gait speed were small. CONCLUSIONS: Gait speed is related to community walking, but an MIC in absolute gait speed could not be determined using a minimally important change on the MFWC or the EDSS as external criteria. PMID- 21622594 TI - Impact of rituximab on relapse rate and disability in neuromyelitis optica. AB - BACKGROUND: Neuromyelitis optica (NMO) is a severe demyelinating disease often leading to serious disability. Accumulating evidence now implicates humoral mechanisms in its pathogenesis. In the absence of an approved therapy, anti inflammatory/immunosuppressant drugs have been used empirically for more than three decades. Recent evidence for a role of antibody to aquaporin-4 in the pathogenesis of NMO has led to the use of rituximab, a monoclonal antibody targeting the CD20 epitope on the entire B cell lineage. OBJECTIVES: To evaluate the impact of rituximab on the relapse rate and disability in NMO. METHODS: This is an IRB approved retrospective longitudinal study of NMO patients treated with rituximab. RESULTS: We identified 53 patients with NMO, 23 of whom had been treated with rituximab. These patients (2 males, 21 females) had a mean age of 37.1 +/- 14.6 years at the time of diagnosis. Eight of the 23 treated with rituximab were treatment naive. All 23 were scheduled to receive infusions every six or 12 months after treatment initiation with a minimum follow-up of six months (median 32.5 months, range 7-63 months). Median relapse rate declined significantly from 1.87 relapses/patient per year to 0.0 relapses/patient per year. Kurtzke Expanded Disability Status Scale (EDSS) scores stabilized or improved in all patients. Use of rituximab is associated with a significant reduction in relapses and disability in patents with NMO. PMID- 21622595 TI - An introduction to item response theory and Rasch models for speech-language pathologists. AB - PURPOSE: To present a primarily conceptual introduction to item response theory (IRT) and Rasch models for speech-language pathologists (SLPs). METHOD: This tutorial introduces SLPs to basic concepts and terminology related to IRT as well as the most common IRT models. The article then continues with an overview of how instruments are developed using IRT and some basic principles of adaptive testing. CONCLUSION: IRT is a set of statistical methods that are increasingly used for developing instruments in speech-language pathology. While IRT is not new, its application in speech-language pathology to date has been relatively limited in scope. Several new IRT-based instruments are currently emerging. IRT differs from traditional methods for test development, typically referred to as classical test theory (CTT), in several theoretical and practical ways. Administration, scoring, and interpretation of IRT instruments are different from methods used for most traditional CTT instruments. SLPs will need to understand the basic concepts of IRT instruments to use these tools in their clinical and research work. This article provides an introduction to IRT concepts drawing on examples from speech-language pathology. PMID- 21622596 TI - Effects of length, complexity, and grammatical correctness on stuttering in Spanish-speaking preschool children. AB - PURPOSE: To explore the effects of utterance length, syntactic complexity, and grammatical correctness on stuttering in the spontaneous speech of young, monolingual Spanish-speaking children. METHOD: Spontaneous speech samples of 11 monolingual Spanish-speaking children who stuttered, ages 35 to 70 months, were examined. Mean number of syllables, total number of clauses, utterance complexity (i.e., containing no clauses, simple clauses, or subordinate and/or conjoined clauses), and grammatical correctness (i.e., the presence or absence of morphological and syntactical errors) in stuttered and fluent utterances were compared. RESULTS: Findings revealed that stuttered utterances in Spanish tended to be longer and more often grammatically incorrect, and contain more clauses, including more subordinate and/or conjoined clauses. However, when controlling for the interrelatedness of syllable number and clause number and complexity, only utterance length and grammatical incorrectness were significant predictors of stuttering in the spontaneous speech of these Spanish-speaking children. Use of complex utterances did not appear to contribute to the prediction of stuttering when controlling for utterance length. CONCLUSIONS: Results from the present study were consistent with many earlier reports of English-speaking children. Both length and grammatical factors appear to affect stuttering in Spanish-speaking children. Grammatical errors, however, served as the greatest predictor of stuttering. PMID- 21622597 TI - African Trypanosoma infection in a dog in France. PMID- 21622598 TI - Mycotic keratoconjunctivitis in 12-day-old red-legged partridges (Alectoris rufa). PMID- 21622599 TI - Distribution of methylene blue injected into the epidural space of anaesthetised sheep. PMID- 21622600 TI - Impact of swallowing on the dynamics of aroma release and perception during the consumption of alcoholic beverages. AB - The consumption protocol used during alcoholic beverage tasting may affect aroma perception. We used an integrated approach combining sensory analysis and physicochemistry to investigate the impact of swallowing on aroma release and perception. A panel of 10 persons evaluated the dynamics of aroma perception during the consumption of a commercial flavored vodka, using the method of temporal dominance of sensations. Two protocols (spitting out or swallowing of the product) were tested. Nosespace analysis was simultaneously carried out by proton transfer reaction mass spectrometry to evaluate aroma release in the nasal cavity. Comparison of the results obtained with the 2 protocols highlighted significant differences in both the perception and the release of aroma: the swallowing of the product resulted in more complex perceptions but decreased the dominance rates of aromatic attributes. Ethanol perception also had an impact when the product was swallowed. Aroma release data partly accounted for the differences in perception, particularly as concerned ethanol release. The time at which dominance appears as well as the dominance duration of some attributes can be related to some temporal parameters of release data. But the lack of knowledge concerning the variety and complexity of mechanisms continues to limit our understanding of relationship between aroma release and perception. PMID- 21622601 TI - Taste perception in honey bees. AB - Taste is crucial for honeybees for choosing profitable food sources, resins, water sources, and for nestmate recognition. Peripheral taste detection occurs within cuticular hairs, the chaetic and basiconic sensilla, which host gustatory receptor cells and, usually a mechanoreceptor cell. Gustatory sensilla are mostly located on the distal segment of the antennae, on the mouthparts, and on the tarsi of the forelegs. These sensilla respond with varying sensitivity to sugars, salts, and possibly amino acids, proteins, and water. So far, no responses of receptor cells to bitter substances were found although inhibitory effects of these substances on sucrose receptor cells could be recorded. When bees are free to express avoidance behaviors, they reject highly concentrated bitter and saline solutions. However, such avoidance disappears when bees are immobilized in the laboratory. In this case, they ingest these solutions, even if they suffer afterward a malaise-like state or even die from such ingestion. Central processing of taste occurs mainly in the subesophageal ganglion, but the nature of this processing remains unknown. We suggest that coding tastants in terms of their hedonic value, thus classifying them in terms of their palatability, is a basic strategy that a central processing of taste should achieve for survival. PMID- 21622602 TI - Proportion of odorants impacts the configural versus elemental perception of a binary blending mixture in newborn rabbits. AB - Processing of odor mixtures by neonates is weakly understood. Previous studies showed that a binary mixture of ethyl isobutyrate/ethyl maltol (odorants A/B) blends in newborn rabbits at the 30/70 ratio: Pups would perceive a configural odor in addition to the components' odors. Here, we investigated whether the emergence of this additional odor in AB is determined by specific ratio(s) of A and B. To that goal, we tested whether pups discriminated between AB mixtures with lower (A(-)B, 8/92 ratio) or higher (A(+)B, 68/32) proportion of A. In Experiment 1, pups conditioned to A (or B) responded to A(-)B and A(+)B but not to AB. In Experiment 2, pups responded to A(-)B after learning of A(-) (and to A(+)B after learning of A(+)) but not to AB. In Experiment 3, after conditioning to A(-)B pups responded to A(-) and B (and to A(+) and B after learning of A(+)B) but not or less to AB. In Experiment 4, pups responded to A(-)B and A(+)B after conditioning to AB. These results confirm the configural perception of certain odor mixtures by young organisms and reveal that the proportion of components is a key factor influencing their coding, recognition, and discrimination of complex stimuli. PMID- 21622606 TI - Working together to tackle zoonoses. PMID- 21622608 TI - New centre aims to strengthen animal welfare education. PMID- 21622611 TI - BSAVA's client leaflets help explain off-label medicines use. PMID- 21622613 TI - Vet reprimanded for self-prescribing. PMID- 21622614 TI - Winning images. PMID- 21622615 TI - Strengthening veterinary involvement in the food chain. PMID- 21622616 TI - Managing metabolic disease and reproductive performance in dairy cattle. PMID- 21622617 TI - Assessment of veterinary practitioners in the British Isles' approaches towards the management of canine osteoarthritis. AB - The aim of this questionnaire-based retrospective study was to ascertain veterinary practitioners in the British Isles' approaches to osteoarthritis in dogs. The Mann-Whitney U test, Kruskal-Wallis test and multiple ordinal logistic regression were used to compare demographic data with treatment options. Questionnaires were returned by 228 practitioners (a response rate of 22.8 per cent). The majority of responses were from males (70 per cent). Eighty-six per cent (188 of 220) of the respondents had graduated from veterinary schools in the UK and Ireland. Veterinarians who had graduated less recently used exercise modulation less frequently (P<0.004, odds ratio [OR]=1.06) and ranked exercise modulation as less important (P=0.008, OR=1.04). Veterinarians who had graduated outside the UK or Ireland were less likely (P=0.033, OR=0.46) to use exercise modulation than those who had graduated in the UK or Ireland. Veterinarians who had graduated more recently were less likely (P=0.008, OR=0.95) to use medications. Female veterinary surgeons were less likely to rank medications as important (P<0.0001, OR=0.29) and less likely to rank corticosteroids as important (P=0.046, OR=0.42) than male veterinary surgeons. Practitioners who had graduated outside the UK or Ireland were less likely (P=0.01, OR=0.30) to rank corticosteroids as important. There was a significant mild negative correlation between the frequency of use of structure-modifying osteoarthritis drugs (SMOADs) and practitioners' opinions on rank importance (-0.32, P<0.0001). Medications such as SMOADs and nutraceuticals were ranked as not important but were used frequently. Overall, age, sex, the university of graduation and the number of canine consultations per day had an impact on the practitioners' treatment protocols. PMID- 21622618 TI - Unusual skin lesions in sheep. PMID- 21622619 TI - National control programme for Salmonella in pigs. PMID- 21622620 TI - Gender balance in the veterinary profession. PMID- 21622621 TI - Joined-up technology? PMID- 21622622 TI - SRSF1 regulates the alternative splicing of caspase 9 via a novel intronic splicing enhancer affecting the chemotherapeutic sensitivity of non-small cell lung cancer cells. AB - Increasing evidence points to the functional importance of alternative splice variations in cancer pathophysiology with the alternative pre-mRNA processing of caspase 9 as one example. In this study, we delve into the underlying molecular mechanisms that regulate the alternative splicing of caspase 9. Specifically, the pre-mRNA sequence of caspase 9 was analyzed for RNA cis-elements known to interact with SRSF1, a required enhancer for caspase 9 RNA splicing. This analysis revealed 13 possible RNA cis-elements for interaction with SRSF1 with mutagenesis of these RNA cis-elements identifying a strong intronic splicing enhancer located in intron 6 (C9-I6/ISE). SRSF1 specifically interacted with this sequence, which was required for SRSF1 to act as a splicing enhancer of the inclusion of the 4 exon cassette. To further determine the biological importance of this mechanism, we employed RNA oligonucleotides to redirect caspase 9 pre mRNA splicing in favor of caspase 9b expression, which resulted in an increase in the IC(50) of non-small cell lung cancer (NSCLC) cells to daunorubicin, cisplatinum, and paclitaxel. In contrast, downregulation of caspase 9b induced a decrease in the IC(50) of these chemotherapeutic drugs. Finally, these studies showed that caspase 9 RNA splicing was a major mechanism for the synergistic effects of combination therapy with daunorubicin and erlotinib. Overall, we have identified a novel intronic splicing enhancer that regulates caspase 9 RNA splicing and specifically interacts with SRSF1. Furthermore, we showed that the alternative splicing of caspase 9 is an important molecular mechanism with therapeutic relevance to NSCLCs. PMID- 21622623 TI - HDAC1 inhibition by maspin abrogates epigenetic silencing of glutathione S transferase pi in prostate carcinoma cells. AB - Both maspin and glutathione S-transferase pi (GSTp) are implicated as tumor suppressors and downregulated in human prostate cancer. It is well established that GSTp downregulation is through DNA methylation-based silencing. We report here that maspin expression in prostate cancer cell line DU145 reversed GSTp DNA methylation, as measured by methylation- specific PCR, MethyLight assay, and bisulfite sequencing. The effect of maspin on GSTp expression was similar to that of the combination of a synthetic histone deacetylase (HDAC) inhibitor and DNA methylation inhibitor 5-aza-2'-deoxycytidine. Maspin expression also led to an increased level of acetylated histone 3, decreased level of methyl transferase, and methyl-CpG-binding domain proteins at the site of demethylated GSTp promoter DNA. Earlier, we have shown that maspin inhibits HDAC1. In PC3 cells, where both maspin and GSTp are expressed at a reduced level, maspin knockdown led to a significant reduction in GSTp expression, whereas dual knockdown of maspin and HDAC1 barely increased the level of GSTp expression. Thus, HDAC1 may play an essential role in cellular response to maspin-mediated GSTp desilencing. Maspin has been shown to increase tumor cell sensitivity to drug-induced apoptosis. Interestingly, GSTp reexpression in the absence of maspin expression perturbation blocked the phosphorylation of histone 2A.X, the induction of hypoxia-induced factor 1alpha (HIF-1alpha), and cell death of LNCaP cells under oxidative stress. Because DNA hypermethylation-based silencing may couple with and depend on histone deacetylation, our study suggests that endogenous HDAC inhibition by maspin may prevent pathologic gene silencing in prostate tumor progression. PMID- 21622624 TI - A novel role of dipeptidyl peptidase 9 in epidermal growth factor signaling. AB - Dipeptidyl peptidase IV (DPP4), DPP8, DPP9, and fibroblast activation protein (FAP), the four proteases of the DPP4 gene family, have unique peptidase and extra-enzymatic activities that have been implicated in various diseases including cancers. We report here a novel role of DPP9 in regulating cell survival and proliferation through modulating molecular signaling cascades. Akt (protein kinase B) activation was significantly inhibited by human DPP9 overexpression in human hepatoma cells (HepG2 and Huh7) and human embryonic kidney cells (HEK293T), whereas extracellular signal-regulated kinases (ERK1/2) activity was unaffected, revealing a pathway-specific effect. Interestingly, the inhibitory effect of DPP9 on Akt pathway activation was growth factor dependent. DPP9 overexpression caused apoptosis and significantly less epidermal growth factor (EGF)-mediated Akt activation in HepG2 cells. However, such inhibitory effect was not observed in cells stimulated with other growth factors, including connective tissue growth factor, hepatic growth factor, insulin or platelet derived growth factor-BB. The effect of DPP9 on Akt did not occur when DPP9 enzyme activity was ablated by either mutagenesis or inhibition. The phosphatidylinositol 3-kinase (PI3K)/Akt pathway is a major downstream effector of Ras. We found that DPP9 and DPP8, but not DPP4 or FAP, associate with H-Ras, a key signal molecule of the EGF receptor signaling pathway. These findings suggest an important signaling role of DPP9 in the regulation of survival and proliferation pathways. PMID- 21622625 TI - Colonic catabolism of ellagitannins, ellagic acid, and raspberry anthocyanins: in vivo and in vitro studies. AB - Red raspberries contain principally anthocyanins and ellagitannins. After ingestion of raspberries by humans, trace levels of anthocyanins, absorbed in the upper gastrointestinal tract, are excreted in urine in amounts corresponding to <0.1% of intake. Urine also contains urolithin-O-glucuronides derived from colonic metabolism of the ellagitannins. Raspberry feedings with ileostomists show that substantial amounts of the anthocyanin and ellagitannin intake are excreted in ileal fluid. In subjects with an intact functioning colon, these compounds would pass to the large intestine. The aim of this study was to identify raspberry-derived phenolic acid catabolites that form in the colon and those that are subsequently excreted in urine. In vitro anaerobic incubation of ellagitannins with fecal suspensions demonstrated conversion to ellagic acid and several urolithins. Fecal suspensions converted 80% of added ellagic acid to urolithins. In vivo, urolithins are excreted in urine as O-glucuronides, not aglycones, indicating that the colonic microflora convert ellagitannins to urolithins, whereas glucuronidation occurs in the wall of the large intestine and/or postabsorption in the liver. Unlike ellagitannins, raspberry anthocyanins were converted in vitro to phenolic acids by anaerobic fecal suspensions. Urinary excretion of phenolic acids after ingestion of raspberries indicates that after formation in the colon some phenolic acids undergo phase II metabolism, resulting in the formation of products that do not accumulate when anthocyanins are degraded in fecal suspensions. There is a growing realization that colonic catabolites such as phenolic acids and urolithins may have important roles in the protective effects of a fruit- and vegetable-rich diet. PMID- 21622626 TI - Sequential metabolism of sesamin by cytochrome P450 and UDP glucuronosyltransferase in human liver. AB - Our previous study revealed that CYP2C9 played a central role in sesamin monocatecholization. In this study, we focused on the metabolism of sesamin monocatechol that was further converted into the dicatechol form by cytochrome P450 (P450) or the glucuronide by UDP-glucuronosyltransferase (UGT). Catecholization of sesamin monocatechol enhances its antioxidant activity, whereas glucuronidation strongly reduces its antioxidant activity. In human liver microsomes, the glucuronidation activity was much higher than the catecholization activity toward sesamin monocatechol. In contrast, in rat liver microsomes, catecholization is predominant over glucuronidation. In addition, rat liver produced two isomers of the glucuronide, whereas human liver produced only one glucuronide. These results suggest a significant species-based difference in the metabolism of sesamin between humans and rats. Kinetic studies using recombinant human UGT isoforms identified UGT2B7 as the most important UGT isoform for glucuronidation of sesamin monocatechol. In addition, a good correlation was observed between the glucuronidation activity and UGT2B7-specific activity in in vitro studies using 10 individual human liver microsomes. These results strongly suggest that UGT2B7 plays an important role in glucuronidation of sesamin monocatechol. Interindividual difference among the 10 human liver microsomes is approximately 2-fold. These results, together with our previous results on the metabolism of sesamin by human P450, suggest a small interindividual difference in sesamin metabolism. We observed the methylation activity toward sesamin monocatechol by catechol O-methyl transferase (COMT) in human liver cytosol. On the basis of these results, we concluded that CYP2C9, UGT2B7, and COMT played essential roles in the metabolism of sesamin in the human liver. PMID- 21622627 TI - In vitro metabolism of the mycotoxin enniatin B in different species and cytochrome p450 enzyme phenotyping by chemical inhibitors. AB - Enniatins are cyclic hexapeptidic mycotoxins produced by fungi growing on field grains, especially in wet climates. They show considerable resistance to food and feed processing technologies and might cause intoxication of humans and animals. Enniatins are also under exploration as anticancer drugs. The observed difference of in vitro and in vivo toxicities suggests low absorption or fast elimination of the enniatins after oral uptake. In the study presented here, in vitro metabolism studies of enniatin B were performed using rat, dog, and human liver microsomes under conditions of linear kinetics to estimate the respective elimination rates. Furthermore, cytochrome P450 reaction phenotyping with chemical inhibitors selective for human enzymes was carried out. Twelve metabolites were separated and characterized by multiple high-performance liquid chromatographic/mass spectrometric analyses as products of oxidation and demethylation reactions. Biotransformation rates and metabolite patterns varied considerably in the three species. The intrinsic clearances determined in assays with rat, dog, and human liver microsomes were 1.16, 8.23, and 1.13 l/(h . kg), respectively. The predicted enniatin B in vivo blood clearances were 1.57 l/(h . kg) in rats, 1.67 l/(h . kg) in dogs, and 0.63 l/(h . kg) in humans. CYP3A4 was important for enniatin B metabolism in human microsomes as shown by 80% inhibition and impaired metabolite formation in the presence of troleandomycin. CYP1A2 and CYP2C19 were additionally involved. Preliminary results showed that CYP3A and CYP1A might also be relevant in rats and dogs. The extensive hepatic metabolism could explain the reduced in vivo potential of enniatin B. PMID- 21622628 TI - Metabolism and disposition of isoflavone conjugated metabolites in humans after ingestion of kinako. AB - Isoflavone aglycones daidzein (Dein) and genistein (Gein) are present primarily as glucuronides and sulfates in human plasma; however, very little is known about the plasma pharmacokinetics of isoflavone conjugates after soy ingestion. The aim of this study was to investigate metabolism and disposition of the isoflavone conjugated metabolites glucuronide or sulfate or both after ingestion of kinako (baked soybean flour) by 10 volunteers. The quantifications of 16 metabolites in plasma and urine were performed by our previously reported high-performance liquid chromatography-UV-diode-array detector method. Plasma concentrations of total Dein and Gein metabolites reached maximal values of 0.64 +/- 0.18 MUM at 4.7 +/- 2.5 h and 1.58 +/- 0.55 MUM at 5.4 +/- 2.1 h, respectively. The area under the curve from 0 to 48 h demonstrated that daidzein-7-glucuronide-4' sulfate (D-7G-4'S) (53.3%) was a major metabolite of Dein and that genistein-7 glucuronide-4'-sulfate (G-7G-4'S) (54.0%) and genistein-4',7-diglucuronide (G 4',7-diG) (26.6%) were major metabolites of Gein in plasma. The compositions of isoflavone metabolites in urine and plasma were greatly different. Approximately half of the 48-h urinary excretion of total Dein metabolites consisted of daidzein-7-glucuronide. The total amounts of genistein-7-glucuronide and genistein-4'-glucuronide were half the total amount of the urinary Gein metabolites. Excretion into urine of D-7G-4'S and G-7G-4'S accounted for only 16% each of the total Dein and Gein metabolites, respectively. The plasma and urine profiles of 16 metabolites of Dein and Gein demonstrate the involvement of desulfation and deglucuronidation of the conjugated metabolites D-7G-4'S, G-7G 4'S, and G-4',7-diG in the process of renal excretion. PMID- 21622629 TI - Integration of QTL and bioinformatic tools to identify candidate genes for triglycerides in mice. AB - To identify genetic loci influencing lipid levels, we performed quantitative trait loci (QTL) analysis between inbred mouse strains MRL/MpJ and SM/J, measuring triglyceride levels at 8 weeks of age in F2 mice fed a chow diet. We identified one significant QTL on chromosome (Chr) 15 and three suggestive QTL on Chrs 2, 7, and 17. We also carried out microarray analysis on the livers of parental strains of 282 F2 mice and used these data to find cis-regulated expression QTL. We then narrowed the list of candidate genes under significant QTL using a "toolbox" of bioinformatic resources, including haplotype analysis; parental strain comparison for gene expression differences and nonsynonymous coding single nucleotide polymorphisms (SNP); cis-regulated eQTL in livers of F2 mice; correlation between gene expression and phenotype; and conditioning of expression on the phenotype. We suggest Slc25a7 as a candidate gene for the Chr 7 QTL and, based on expression differences, five genes (Polr3 h, Cyp2d22, Cyp2d26, Tspo, and Ttll12) as candidate genes for Chr 15 QTL. This study shows how bioinformatics can be used effectively to reduce candidate gene lists for QTL related to complex traits. PMID- 21622630 TI - ApoA-I deficiency in mice is associated with redistribution of apoA-II and aggravated AApoAII amyloidosis. AB - Apolipoprotein A-II (apoA-II) is the second major apolipoprotein following apolipoprotein A-I (apoA-I) in HDL. ApoA-II has multiple physiological functions and can form senile amyloid fibrils (AApoAII) in mice. Most circulating apoA-II is present in lipoprotein A-I/A-II. To study the influence of apoA-I on apoA-II and AApoAII amyloidosis, apoA-I-deficient (C57BL/6J.Apoa1-/-) mice were used. Apoa1-/- mice showed the expected significant reduction in total cholesterol (TC), HDL cholesterol (HDL-C), and triglyceride (TG) plasma levels. Unexpectedly, we found that apoA-I deficiency led to redistribution of apoA-II in HDL and an age-related increase in apoA-II levels, accompanied by larger HDL particle size and an age-related increase in TC, HDL-C, and TG. Aggravated AApoAII amyloidosis was induced in Apoa1-/- mice systemically, especially in the heart. These results indicate that apoA-I plays key roles in maintaining apoA-II distribution and HDL particle size. Furthermore, apoA-II redistribution may be the main reason for aggravated AApoAII amyloidosis in Apoa1-/- mice. These results may shed new light on the relationship between apoA-I and apoA-II as well as provide new information concerning amyloidosis mechanism and therapy. PMID- 21622631 TI - Reliability and validity of the INFOOT three-dimensional foot digitizer for patients with rheumatoid arthritis. AB - BACKGROUND: Abnormal foot posture and deformities are identified as important features in rheumatoid arthritis. There is still no consensus regarding the optimum technique(s) for quantifying these features; hence, a foot digitizer might be used as an objective measurement tool. We sought to assess the validity and reliability of the INFOOT digitizer. METHODS: To investigate the validity of the INFOOT digitizer compared with clinical measurements, we calculated Pearson correlation coefficients. To investigate the reliability of the INFOOT digitizer, we calculated intraclass correlation coefficients, SEMs, smallest detectable differences, and smallest detectable difference percentages. RESULTS: Most of the 38 parameters showed good intraclass correlation coefficients, with values greater than 0.9 for 30 parameters and greater than 0.8 for seven parameters. The left heel bone angle expressed a moderate correlation, with a value of 0.609. The SEM values varied between 0.31 and 3.51 mm for the length and width measures, between 0.74 and 5.58 mm for the height data, between 0.75 and 5.9 mm for the circumferences, and between 0.78 degrees and 2.98 degrees for the angles. The smallest detectable difference values ranged from 0.86 to 16.36 mm for length, width, height, and circumference measures and from 2.17 degrees to 8.26 degrees for the angle measures. For the validity of the INFOOT three-dimensional foot digitizer, Pearson correlation coefficients varied between 0.750 and 0.997. CONCLUSIONS: In this rheumatoid arthritis population, good validity was demonstrated compared with clinical measurements, and most of the obtained parameters proved to be reliable. PMID- 21622632 TI - Foot biomechanics in patients with diabetes mellitus: doubts regarding the relationship between neuropathy, foot motion, and deformities. AB - BACKGROUND: We sought to identify the biomechanical characteristics of the feet of patients with diabetes mellitus and the interrelationship with diabetic neuropathy by determining the range of joint mobility and the presence and locations of calluses and foot deformities. METHODS: This observational comparative study involved 281 patients with diabetes mellitus who underwent neurologic and vascular examinations. Joint mobility studies were performed, and deformities and hyperkeratosis locations were assessed. RESULTS: No substantial differences were found between patients with and without neuropathy in joint mobility range. Neuropathy was seen as a risk factor only in the passive range of motion of the first metatarsophalangeal joint (mean +/- SD: 57.2 degrees +/- 19.5 degrees versus 50.3 degrees +/- 22.5 degrees , P = .008). Mean +/- SD ankle joint mobility values were similar in both groups (83.0 degrees +/- 5.2 degrees versus 82.8 degrees +/- 9.3 degrees , P = .826). Patients without neuropathy had a higher rate of foot deformities such as hallux abductus valgus and hammer toes. There was also a higher presence of calluses in patients without neuropathy (82.8% versus 72.6%; P = .039). CONCLUSIONS: Diabetic neuropathy was not related to limited joint mobility and the presence of calluses. Patients with neuropathy did not show a higher risk of any of the deformities examined. These findings suggest that the etiology of biomechanical alterations in diabetic people is complex and may involve several anatomically and pathologically predisposing factors. PMID- 21622633 TI - Patellofemoral pain syndrome and its association with hip, ankle, and foot function in 16- to 18-year-old high school students: a single-blind case-control study. AB - BACKGROUND: An increased pronated foot posture is believed to contribute to patellofemoral pain syndrome (PFPS), but the relationship between these phenomena is still controversial. The objectives of this study were to investigate the prevalence of PFPS in high school students and to compare passive internal and external hip rotation, passive dorsiflexion, and navicular drop and drift between healthy high school students and students with PFPS. METHODS: All 16- to 18-year old students in a Danish high school were invited to join this single-blind case control study (N = 299). All of the students received a questionnaire regarding knee pain. The main outcome measurements were prevalence of PFPS, navicular drop and drift, passive ankle dorsiflexion, passive hip rotation in the prone position, and activity level. The case group consisted of all students with PFPS. From the same population, a randomly chosen control group was formed. RESULTS: The prevalence of knee pain was 25%. Of the 24 students with knee pain, 13 were diagnosed as having PFPS. This corresponds to a PFPS prevalence of 6%. Mean navicular drop and drift were higher in the PFPS group versus the control group (navicular drop: 4.2 mm [95% confidence interval (CI), 3.2-5.3 mm] versus 2.9 mm [95% CI, 2.5-3.3 mm]; and navicular drift: 2.6 mm [95% CI, 1.6-3.7 mm] versus 1.4 mm [95% CI, 0.9-2.0 mm]). Higher passive ankle dorsiflexion was also identified in the PFPS group (22.2 degrees [95% CI, 18 degrees -26 degrees ] versus 17.7 degrees [95% CI, 15 degrees -20 degrees ]). CONCLUSIONS: This study demonstrated greater navicular drop, navicular drift, and dorsiflexion in high school students with PFPS compared with healthy students and highlights that foot posture is important to consider as a factor where patients with PFPS diverge from healthy individuals. PMID- 21622634 TI - Reducing the pain of local 1% lidocaine infiltration with a preceding bacteriostatic saline injection: a double-blind prospective trial. AB - BACKGROUND: Lidocaine injection for local anesthesia is a common podiatric medical procedure. We tested the hypothesis that injection of bacteriostatic saline solution containing 0.9% benzyl alcohol before the lidocaine infiltration can reduce the burning caused by lidocaine injection. METHODS: This double-blind prospective trial involved 45 participants who each received four injections in two areas of the dorsum of the foot and rated the perceived pain on a visual analog scale. The order of the injections was designed to disguise the control and intervention arms of the study. RESULTS: The sensation of the lidocaine injection after the injection of saline was reduced significantly (P = .028). The percentage of lidocaine injections with visual analog scale scores of 0 increased by 36% after preinjection with bacteriostatic saline solution containing 0.9% benzyl alcohol. CONCLUSIONS: The fact that 40% of the intervention visual analog scale pain scores for lidocaine injections were 0 suggests that a near painless lidocaine injection technique is an achievable goal and that the present technique is a simple and inexpensive method of reducing the pain of lidocaine injections. PMID- 21622635 TI - Barefoot running claims and controversies: a review of the literature. AB - BACKGROUND: Barefoot running is slowly gaining a dedicated following. Proponents of barefoot running claim many benefits, such as improved performance and reduced injuries, whereas detractors warn of the imminent risks involved. METHODS: Multiple publications were reviewed using key words. RESULTS: A review of the literature uncovered many studies that have looked at the barefoot condition and found notable differences in gait and other parameters. These findings, along with much anecdotal information, can lead one to extrapolate that barefoot runners should have fewer injuries, better performance, or both. Several athletic shoe companies have designed running shoes that attempt to mimic the barefoot condition and, thus, garner the purported benefits of barefoot running. CONCLUSIONS: Although there is no evidence that either confirms or refutes improved performance and reduced injuries in barefoot runners, many of the claimed disadvantages to barefoot running are not supported by the literature. Nonetheless, it seems that barefoot running may be an acceptable training method for athletes and coaches who understand and can minimize the risks. PMID- 21622636 TI - A quantitative method for measuring forces applied by nail braces. AB - BACKGROUND: Nail bracing is a conservative method used for ingrown nails; however, lack of objective measurements limits its use for various nails. METHODS: Double-string nail braces with extra metal springs were applied to 12 patients with 21 chronic, thick, and overcurved ingrown nails. Force was measured with a force gauge meter. Treatment was stopped once patients stood on their tiptoes and walked in shoes pain free without braces. A force gauge meter was also used on a model nail to show the forces applied by various nail braces and to compare their pulling forces. RESULTS: After 6 to 10 months of treatment, all of the patients were pain free; 600 to 1,000 centi Newtons of force were applied to the nails. As the width of the nail increased, so did the force. CONCLUSIONS: Braces exert more force on larger nails, which may shorten treatment durations. By measuring forces, it may be possible to standardize force and duration of treatment according to variables such as nail thickness, nail width, angle of ingrown nail, and duration of symptoms. PMID- 21622637 TI - Pigmented villonodular synovitis of the ankle: radiologic characteristics. AB - BACKGROUND: Pigmented villonodular synovitis (PVNS) of the ankle is a rare benign proliferative growth of the synovium. Studies of the radiologic characteristics of ankle PVNS are sparse. METHODS: To characterize the radiologic features of ankle PVNS, five patients with histologically proven ankle PVNS were retrospectively studied. The features of their radiographs, computed tomographic scans, and magnetic resonance images were reviewed, with emphasis on the morphological features, extension, margin, bone involvement, signal intensity, and degree of magnetic resonance enhancement. RESULTS: All five lesions were diffuse, affecting the ankle and distal tibiofibular joint; three lesions also involved the subtalar joint. Radiography demonstrated extrinsic bone erosions with marginal sclerosis of the involved joints in all of the patients, but computed tomography identified this much better than did radiography. Magnetic resonance imaging revealed multiple lobulated soft-tissue masses in all of the cases. These soft-tissue masses surrounded the flexor hallux longus tendon and were hypointense on T1-weighted images, with a heterogeneous signal in two cases and homogenous hypointensity in three cases on fat-suppressed T2-weighted images. In one patient who underwent gadolinium-enhanced imaging, the masses showed intense enhancement. CONCLUSIONS: Magnetic resonance imaging is the best way to reveal ankle PVNS. Magnetic resonance imaging findings of predominant hypointensity on all pulse sequences and standard radiography findings of bone erosion with marginal sclerosis are characteristic. PMID- 21622638 TI - Management in the wound-care center outpatient setting of a diabetic patient with forefoot osteomyelitis using Cerament Bone Void Filler impregnated with vancomycin: off-label use. AB - Several nonbiodegradable and biodegradable antibiotic cement delivery systems are available for the delivery of antibiotics for adjunctive therapy in the management of osteomyelitis. A major nonbiodegradable delivery system is polymethylmethacrylate beads. Antibiotics that can be incorporated into this delivery system are limited to the heat-stable antibiotics vancomycin and aminoglycosides, tobramycin being the most popular. Calcium sulfate and hydroxyapatite (Cerament Bone Void Filler) is a unique biocompatible and biodegradable ceramic bone void filler that can successfully deliver heat-stable and heat-unstable antibiotics in musculoskeletal infections. The use of Cerament as antibiotic beads has not been previously reported. An off-label case of diabetic foot osteomyelitis successfully managed with surgical bone resection and vancomycin Cerament antibiotic beads is presented. Subsequent surgery for the bone infection and staged removal of the antibiotic beads was not necessary. PMID- 21622639 TI - Metastatic renal cell carcinoma presenting as foot metastasis: case report and review of the literature. AB - Malignant tumors frequently metastasize to bone centrally in the skeleton. Metastatic disease distal to the knee is unusual. Metastasis to the foot (acrometastasis) is rare (0.01%) and is usually a late manifestation of disseminated disease. The purpose of this article is to present a rare case of metastatic renal cell carcinoma with foot metastasis as the primary manifestation along with another rare localization of metastatic disease distal to the knee, in the contralateral tibial diaphysis. To highlight the delay in diagnosis of such a rare condition to consider it in the diagnosis of a painful foot, we also present a review of the literature. PMID- 21622640 TI - An unusual cause of a cystic lesion with an osteochondral defect in the talus: intraosseous lipoma. AB - An intraosseous lipoma is a rare benign bone lesion that proliferates from mature lipocytes. It occurs most frequently in the lower limb, particularly in the calcaneus. The talus is an unusual location for this rare lesion. A review of the literature produced only two reports with talar intraosseous lipomas under the name of intraosseous lipomatosis, which described multiple lipomas in different areas. We describe a 38-year-old male patient who had an isolated intraosseous lipoma with an osteochondral defect in the talus and was treated with autologous osteochondral graft transplantation by medial malleolar osteotomy. He could walk with full weightbearing without any assistance at the end of 12 months. Intraosseous lipoma localized in the talus may be confused radiologically with other bone lesions, especially with unicameral bone cyst, if it is associated with an osteochondral defect. Autologous osteochondral graft transplantation is a successful treatment method for talar intraosseous lipoma. PMID- 21622641 TI - Human digit partially consumed by a canine during sleep in a patient with neuropathy and diabetes. AB - A traumatic amputation of a digit as a result of canine mastication and ingestion occurred in a 48-year-old woman with type 2 diabetes and peripheral neuropathy. The injury occurred during sleep and was not felt by the patient. The dangers of sleeping with one's canine for those with neuropathic wounds are presented, and the literature is reviewed. PMID- 21622642 TI - Curation of characterized glycoside hydrolases of fungal origin. AB - Fungi produce a wide range of extracellular enzymes to break down plant cell walls, which are composed mainly of cellulose, lignin and hemicellulose. Among them are the glycoside hydrolases (GH), the largest and most diverse family of enzymes active on these substrates. To facilitate research and development of enzymes for the conversion of cell-wall polysaccharides into fermentable sugars, we have manually curated a comprehensive set of characterized fungal glycoside hydrolases. Characterized glycoside hydrolases were retrieved from protein and enzyme databases, as well as literature repositories. A total of 453 characterized glycoside hydrolases have been cataloged. They come from 131 different fungal species, most of which belong to the phylum Ascomycota. These enzymes represent 46 different GH activities and cover 44 of the 115 CAZy GH families. In addition to enzyme source and enzyme family, available biochemical properties such as temperature and pH optima, specific activity, kinetic parameters and substrate specificities were recorded. To simplify comparative studies, enzyme and species abbreviations have been standardized, Gene Ontology terms assigned and reference to supporting evidence provided. The annotated genes have been organized in a searchable, online database called mycoCLAP (Characterized Lignocellulose-Active Proteins of fungal origin). It is anticipated that this manually curated collection of biochemically characterized fungal proteins will be used to enhance functional annotation of novel GH genes. Database URL: http://mycoCLAP.fungalgenomics.ca/. PMID- 21622643 TI - Determination of a practical pH cutoff level for reliable confirmation of nasogastric tube placement. AB - BACKGROUND: Enteral feeding is a common method of nutrition support when oral intake is inadequate. Confirmation of correct nasogastric (NG) tube placement is essential. Risks of morbidity/mortality associated with misplacement in the lung are well documented. Studies indicate that pH <= 4 confirms gastric aspirate, but in pediatrics, a pH of gastric aspirate is often >4. The goal of this study was to determine a reliable and practical pH value to confirm NG tube placement, without increasing the risk of not identifying a misplaced NG tube. METHODS: Pediatric inpatients older than 4 weeks receiving enteral nutrition (nasogastric or gastrostomy) were recruited over 9 months. Aspirate samples were pH tested at NG tube placement and before feedings. If pH >4, NG tube position was confirmed by chest radiograph or further investigations. In addition, intensive care unit (ICU) patients who required endotracheal suctioning were recruited, and endotracheal aspirate samples were pH tested. RESULTS: A total of 4,330 gastric aspirate samples (96% nasogastric) were collected from 645 patients with a median (interquartile range [IQR]) age of 1.0 years (0.3-5.2 years). The mean (standard deviation [SD]) pH of these gastric samples was 3.6 (1.4) (range, 0-9). pH was >4 in 1,339 (30.9%) gastric aspirate samples, and of these, 244 were radiographed, which identified 10 misplaced tubes (1 with pH 5.5). A total of 65 endotracheal aspirate samples were collected from 19 ICU patients with a median (IQR) age of 0.6 years (0.4-5.2 years). The mean (SD) pH of these samples was 8.4 (0.8) (range, 6-9.5). CONCLUSION: Given that the lowest pH value of endotracheal aspirate sample was 6, and a misplaced NG tube was identified with pH 5.5, it is proposed that a gastric aspirate pH <= 5 is a safer, reliable, and practical cutoff in this population. PMID- 21622644 TI - Comparison of thalidomide and lenalidomide as therapy for myelofibrosis. AB - With the use of the International Working Group for Myelofibrosis Treatment and Research consensus criteria, we re-assessed the efficacy of thalidomide and lenalidomide in 125 patients with myelofibrosis treated in 3 consecutive phase 2 trials: 44 received single-agent thalidomide, 41 single-agent lenalidomide, and 40 a combination of lenalidomide plus prednisone. The thalidomide group included significantly more untreated patients and patients with performance status of 2. The Lenalidomide-based therapy produced higher efficacy (34%-38%) than thalidomide (16%; P = .06). Responses to thalidomide were seen within 3-15 weeks, whereas responses to the lenalidomide-based therapy were also seen after a prolonged course of therapy (range, 2-45 weeks). Lenalidomide plus prednisone therapy resulted in significantly longer response duration (median, 34 months) than single-agent lenalidomide or thalidomide (median, 7 and 13 months, respectively; P = .042). Fewer patients (P = .001) discontinued the lenalidomide plus prednisone therapy (13%) because of side effects then patients on single agents therapy (32%-39%). In conclusion, the combination of lenalidomide plus prednisone appears to be more effective and safer than single-agent thalidomide or lenalidomide. PMID- 21622645 TI - Transcription factor networks in erythroid cell and megakaryocyte development. AB - Erythroid cells and megakaryocytes are derived from a common precursor, the megakaryocyte-erythroid progenitor. Although these 2 closely related hematopoietic cell types share many transcription factors, there are several key differences in their regulatory networks that lead to differential gene expression downstream of the megakaryocyte-erythroid progenitor. With the advent of next-generation sequencing and our ability to precisely define transcription factor chromatin occupancy in vivo on a global scale, we are much closer to understanding how these 2 lineages are specified and in general how transcription factor complexes govern hematopoiesis. PMID- 21622646 TI - Loss of p19Arf in a Rag1(-/-) B-cell precursor population initiates acute B lymphoblastic leukemia. AB - In human B-acute lymphoblastic leukemia (B-ALL), RAG1-induced genomic alterations are important for disease progression. However, given that biallelic loss of the RAG1 locus is observed in a subset of cases, RAG1's role in the development of B ALL remains unclear. We chose a p19Arf(-/-)Rag1(-/-) mouse model to confirm the previously published results concerning the contribution of CDKN2A (p19ARF /INK4a) and RAG1 copy number alterations in precursor B cells to the initiation and/or progression to B-acute lymphoblastic leukemia (B-ALL). In this murine model, we identified a new, Rag1-independent leukemia-initiating mechanism originating from a Sca1(+)CD19(+) precursor cell population and showed that Notch1 expression accelerates the cells' self-renewal capacity in vitro. In human RAG1-deficient BM, a similar CD34(+)CD19(+) population expressed p19ARF. These findings suggest that combined loss of p19Arf and Rag1 results in B-cell precursor leukemia in mice and may contribute to the progression of precursor B ALL in humans. PMID- 21622647 TI - New mechanism of X-linked anhidrotic ectodermal dysplasia with immunodeficiency: impairment of ubiquitin binding despite normal folding of NEMO protein. AB - Nuclear factor-kappaB essential modulator (NEMO), the regulatory subunit of the IkappaB kinase complex, is a critical component of the NF-kappaB pathway. Hypomorphic mutations in the X-linked human NEMO gene cause various forms of anhidrotic ectodermal dysplasia with immunodeficiency (EDA-ID). All known X linked EDA-ID-causing mutations impair NEMO protein expression, folding, or both. We describe here 2 EDA-ID-causing missense mutations that affect the same residue in the CC2-LZ domain (D311N and D311G) that do not impair NEMO production or folding. Structural studies based on pull-down experiments showed a defect in noncovalent interaction with K63-linked and linear polyubiquitin chains for these mutant proteins. Functional studies on the patients' cells showed an impairment of the classic NF-kappaB signaling pathways after activation of 2 NEMO ubiquitin binding-dependent receptors, the TNF and IL-1beta receptors, and in the CD40 dependent NF-kappaB pathway. We report the first human NEMO mutations responsible for X-linked EDA-ID found to affect the polyubiquitin binding of NEMO rather than its expression and folding. These experiments demonstrate that the binding of human NEMO to polyubiquitin is essential for NF-kappaB activation. They also demonstrate that the normal expression and folding of NEMO do not exclude a pathogenic role for NEMO mutations in patients with EDA-ID. PMID- 21622648 TI - Quantitative immunofluorescence mapping reveals little functional coclustering of proteins within platelet alpha-granules. AB - Platelets are small anucleate blood cells that aggregate to seal leaks at sites of vascular injury and are important in the pathology of atherosclerosis, acute coronary syndromes, rheumatoid arthritis, cancer, and the regulation of angiogenesis. In all cases, platelet aggregation requires release of stored proteins from alpha-granules. However, how proteins with potentially antagonistic functions are packaged within alpha-granules is controversial. One possibility is the packaging of functional agonists and antagonists into different alpha-granule populations. By quantitative immunofluorescence colocalization, we found that pair-wise comparisons of 15 angiogenic-relevant alpha-granule proteins displayed little, if any, pattern of functional coclustering. Rather, the data suggested a Gaussian distribution indicative of stochastic protein delivery to individual granules. The apparent physiologic paradox raised by these data may be explained through alternate mechanisms, such as differential content release through incomplete granule fusion or dampened and balanced regulatory networks brought about by the corelease of antagonistic factors. PMID- 21622649 TI - Adulthood residential ultraviolet radiation, sun sensitivity, dietary vitamin D, and risk of lymphoid malignancies in the California Teachers Study. AB - To lend clarity to inconsistent prior findings of an inverse association between ultraviolet radiation (UVR) exposure and risk of lymphoid malignancies, we examined the association of prospectively ascertained residential ambient UVR exposure with risk of non-Hodgkin lymphomas (NHLs), multiple myeloma (MM), and classical Hodgkin lymphoma in the California Teachers Study cohort. Among 121 216 eligible women, 629 were diagnosed with NHL, 119 with MM, and 38 with Hodgkin lymphoma between 1995-1996 and 2007. Cox proportional hazards regression was used to estimate incidence rate ratios (RRs) with 95% confidence intervals (CIs). Residential UVR levels within a 20-km radius were associated with reduced risk of overall NHL (RR for highest vs lowest statewide quartile of minimum UVR [>= 5100 vs < 4915 W-h/m(2)], 0.58; 95% CI, 0.42-0.80), especially diffuse large B-cell lymphoma (RR, 0.36; 95% CI, 0.17-0.78) and chronic lymphocytic leukemia/small lymphocytic lymphoma (RR, 0.46; 95% CI, 0.21-1.01), and MM (RR for maximum UVR, 0.57; 95% CI, 0.36-0.90). These associations were not modified by skin sensitivity to sunlight, race/ethnicity, body mass index, or neighborhood socioeconomic status. Dietary vitamin D also was not associated with risk of lymphoid malignancies. These results support a protective effect of routine residential UVR exposure against lymphomagenesis through mechanisms possibly independent of vitamin D. PMID- 21622650 TI - Dismal prognostic value of monosomal karyotype in elderly patients with acute myeloid leukemia: a GOELAMS study of 186 patients with unfavorable cytogenetic abnormalities. AB - The prognosis of acute myeloid leukemia (AML) is very poor in elderly patients, especially in those classically defined as having unfavorable cytogenetics. The recent monosomal karyotype (MK) entity, defined as 2 or more autosomal monosomies or combination of 1 monosomy with structural abnormalities, has been reported to be associated with a worse outcome than the traditional complex karyotype (CK). In this retrospective study of 186 AML patients older than 60 years, the prognostic influence of MK was used to further stratify elderly patients with unfavorable cytogenetics. CK was observed in 129 patients (69%), and 110 exhibited abnormalities according to the definition of MK (59%). MK(+) patients had a complete response rate significantly lower than MK(-) patients: 37% vs 64% (P = .0008), and their 2-year overall survival was also decreased at 7% vs 22% (P < .0001). In multivariate analysis, MK appeared as the major independent prognostic factor related to complete remission achievement (odds ratio = 2.3; 95% confidence interval, 1-5.4, P = .05) and survival (hazard ratio = 1.7; 95% confidence interval, 1.1-2.5, P = .008). In the subgroup of 129 CK(+) patients, survival was dramatically decreased for MK(+) patients (8% vs 28% at P = .03). These results demonstrate that MK is a major independent factor of very poor prognosis in elderly AML. PMID- 21622651 TI - Stromal cell-derived factor-1 and hematopoietic cell homing in an adult zebrafish model of hematopoietic cell transplantation. AB - In mammals, stromal cell-derived factor-1 (SDF-1) promotes hematopoietic cell mobilization and migration. Although the zebrafish, Danio rerio, is an emerging model for studying hematopoietic cell transplantation (HCT), the role of SDF-1 in the adult zebrafish has yet to be determined. We sought to characterize sdf-1 expression and function in the adult zebrafish in the context of HCT. In situ hybridization of adult zebrafish organs shows sdf-1 expression in kidney tubules, gills, and skin. Radiation up-regulates sdf-1 expression in kidney to nearly 4 fold after 40 Gy. Assays indicate that zebrafish hematopoietic cells migrate toward sdf-1, with a migration ratio approaching 1.5 in vitro. A sdf-1a:DsRed2 transgenic zebrafish allows in vivo detection of sdf-1a expression in the adult zebrafish. Matings with transgenic reporters localized sdf-1a expression to the putative hematopoietic cell niche in proximal and distal renal tubules and collecting ducts. Importantly, transplant of hematopoietic cells into myelosuppressed recipients indicated migration of hematopoietic cells to sdf-1a expressing sites in the kidney and skin. We conclude that sdf-1 expression and function in the adult zebrafish have important similarities to mammals, and this sdf-1 transgenic vertebrate will be useful in characterizing the hematopoietic cell niche and its interactions with hematopoietic cells. PMID- 21622652 TI - Regulation of TMPRSS6 by BMP6 and iron in human cells and mice. AB - Mutations in transmembrane protease, serine 6 (TMPRSS6), encoding matriptase-2, are responsible for the familial anemia disorder iron-refractory iron deficiency anemia (IRIDA). Patients with IRIDA have inappropriately elevated levels of the iron regulatory hormone hepcidin, suggesting that TMPRSS6 is involved in negatively regulating hepcidin expression. Hepcidin is positively regulated by iron via the bone morphogenetic protein (BMP)-SMAD signaling pathway. In this study, we investigated whether BMP6 and iron also regulate TMPRSS6 expression. Here we demonstrate that, in vitro, treatment with BMP6 stimulates TMPRSS6 expression at the mRNA and protein levels and leads to an increase in matriptase 2 activity. Moreover, we identify that inhibitor of DNA binding 1 is the key element of the BMP-SMAD pathway to regulate TMPRSS6 expression in response to BMP6 treatment. Finally, we show that, in mice, Tmprss6 mRNA expression is stimulated by chronic iron treatment or BMP6 injection and is blocked by injection of neutralizing antibody against BMP6. Our results indicate that BMP6 and iron not only induce hepcidin expression but also induce TMPRSS6, a negative regulator of hepcidin expression. Modulation of TMPRSS6 expression could serve as a negative feedback inhibitor to avoid excessive hepcidin increases by iron to help maintain tight homeostatic balance of systemic iron levels. PMID- 21622653 TI - How I treat mucormycosis. AB - Unlike invasive aspergillosis, the prognosis and outcome of hematologic malignancy patients who develop invasive mucormycosis have not significantly improved over the past decade as a majority of patients who develop the infection still die 12 weeks after diagnosis. However, early recognition and treatment of invasive mucormycosis syndromes, as well as individualized approaches to treatment and secondary prophylaxis, could improve the odds of survival, even in the most persistently immunosuppressed patient receiving chemotherapy and/or of stem cell transplantation. Herein, we describe the subtle clinical and radiographic clues that should alert the hematologist to the possibility of mucormycosis, and aggressive and timely treatment approaches that may limit the spread of infection before it becomes fatal. Hematology patients with this opportunistic infection require integrated care across several disciplines and frequently highly individualized and complex sequence of decision-making. We also offer perspectives for the use of 2 antifungals, amphotericin B products and posaconazole, with activity against Mucorales. The availability of posaconazole in an oral formulation that can be administered safely for prolonged periods makes it an attractive agent for long-term primary and secondary prophylaxis. However, serum drug concentration monitoring may be required to minimize breakthrough infection or relapsing mucormycosis associated with inadequate blood concentrations. PMID- 21622654 TI - SLiMSearch 2.0: biological context for short linear motifs in proteins. AB - Short, linear motifs (SLiMs) play a critical role in many biological processes. The SLiMSearch 2.0 (Short, Linear Motif Search) web server allows researchers to identify occurrences of a user-defined SLiM in a proteome, using conservation and protein disorder context statistics to rank occurrences. User-friendly output and visualizations of motif context allow the user to quickly gain insight into the validity of a putatively functional motif occurrence. For each motif occurrence, overlapping UniProt features and annotated SLiMs are displayed. Visualization also includes annotated multiple sequence alignments surrounding each occurrence, showing conservation and protein disorder statistics in addition to known and predicted SLiMs, protein domains and known post-translational modifications. In addition, enrichment of Gene Ontology terms and protein interaction partners are provided as indicators of possible motif function. All web server results are available for download. Users can search motifs against the human proteome or a subset thereof defined by Uniprot accession numbers or GO term. The SLiMSearch server is available at: http://bioware.ucd.ie/slimsearch2.html. PMID- 21622655 TI - Phosfinder: a web server for the identification of phosphate-binding sites on protein structures. AB - Phosfinder is a web server for the identification of phosphate binding sites in protein structures. Phosfinder uses a structural comparison algorithm to scan a query structure against a set of known 3D phosphate binding motifs. Whenever a structural similarity between the query protein and a phosphate binding motif is detected, the phosphate bound by the known motif is added to the protein structure thus representing a putative phosphate binding site. Predicted binding sites are then evaluated according to (i) their position with respect to the query protein solvent-excluded surface and (ii) the conservation of the binding residues in the protein family. The server accepts as input either the PDB code of the protein to be analyzed or a user-submitted structure in PDB format. All the search parameters are user modifiable. Phosfinder outputs a list of predicted binding sites with detailed information about their structural similarity with known phosphate binding motifs, and the conservation of the residues involved. A graphical applet allows the user to visualize the predicted binding sites on the query protein structure. The results on a set of 52 apo/holo structure pairs show that the performance of our method is largely unaffected by ligand-induced conformational changes. Phosfinder is available at http://phosfinder.bio.uniroma2.it. PMID- 21622656 TI - CoMet--a web server for comparative functional profiling of metagenomes. AB - Analyzing the functional potential of newly sequenced genomes and metagenomes has become a common task in biomedical and biological research. With the advent of high-throughput sequencing technologies comparative metagenomics opens the way to elucidate the genetically determined similarities and differences of complex microbial communities. We developed the web server 'CoMet' (http://comet.gobics.de), which provides an easy-to-use comparative metagenomics platform that is well-suitable for the analysis of large collections of metagenomic short read data. CoMet combines the ORF finding and subsequent assignment of protein sequences to Pfam domain families with a comparative statistical analysis. Besides comprehensive tabular data files, the CoMet server also provides visually interpretable output in terms of hierarchical clustering and multi-dimensional scaling plots and thus allows a quick overview of a given set of metagenomic samples. PMID- 21622657 TI - BAR-PLUS: the Bologna Annotation Resource Plus for functional and structural annotation of protein sequences. AB - We introduce BAR-PLUS (BAR(+)), a web server for functional and structural annotation of protein sequences. BAR(+) is based on a large-scale genome cross comparison and a non-hierarchical clustering procedure characterized by a metric that ensures a reliable transfer of features within clusters. In this version, the method takes advantage of a large-scale pairwise sequence comparison of 13,495,736 protein chains also including 988 complete proteomes. Available sequence annotation is derived from UniProtKB, GO, Pfam and PDB. When PDB templates are present within a cluster (with or without their SCOP classification), profile Hidden Markov Models (HMMs) are computed on the basis of sequence to structure alignment and are cluster-associated (Cluster-HMM). Therefrom, a library of 10,858 HMMs is made available for aligning even distantly related sequences for structural modelling. The server also provides pairwise query sequence-structural target alignments computed from the correspondent Cluster-HMM. BAR(+) in its present version allows three main categories of annotation: PDB [with or without SCOP (*)] and GO and/or Pfam; PDB (*) without GO and/or Pfam; GO and/or Pfam without PDB (*) and no annotation. Each category can further comprise clusters where GO and Pfam functional annotations are or are not statistically significant. BAR(+) is available at http://bar.biocomp.unibo.it/bar2.0. PMID- 21622658 TI - Roles of DNA polymerase I in leading and lagging-strand replication defined by a high-resolution mutation footprint of ColE1 plasmid replication. AB - DNA polymerase I (pol I) processes RNA primers during lagging-strand synthesis and fills small gaps during DNA repair reactions. However, it is unclear how pol I and pol III work together during replication and repair or how extensive pol I processing of Okazaki fragments is in vivo. Here, we address these questions by analyzing pol I mutations generated through error-prone replication of ColE1 plasmids. The data were obtained by direct sequencing, allowing an accurate determination of the mutation spectrum and distribution. Pol I's mutational footprint suggests: (i) during leading-strand replication pol I is gradually replaced by pol III over at least 1.3 kb; (ii) pol I processing of Okazaki fragments is limited to ~20 nt and (iii) the size of Okazaki fragments is short (~250 nt). While based on ColE1 plasmid replication, our findings are likely relevant to other pol I replicative processes such as chromosomal replication and DNA repair, which differ from ColE1 replication mostly at the recruitment steps. This mutation footprinting approach should help establish the role of other prokaryotic or eukaryotic polymerases in vivo, and provides a tool to investigate how sequence topology, DNA damage, or interactions with protein partners may affect the function of individual DNA polymerases. PMID- 21622659 TI - The in vitro loose dimer structure and rearrangements of the HIV-2 leader RNA. AB - RNA dimerization is an essential step in the retroviral life cycle. Dimerization and encapsidation signals, closely linked in HIV-2, are located in the leader RNA region. The SL1 motif and nucleocapsid protein are considered important for both processes. In this study, we show the structure of the HIV-2 leader RNA (+1-560) captured as a loose dimer. Potential structural rearrangements within the leader RNA were studied. In the loose dimer form, the HIV-2 leader RNA strand exists in vitro as a single global fold. Two kissing loop interfaces within the loose dimer were identified: SL1/SL1 and TAR/TAR. Evidence for these findings is provided by RNA probing using SHAPE, chemical reagents, enzymes, non-denaturing PAGE mobility assays, antisense oligonucleotides hybridization and analysis of an RNA mutant. Both TAR and SL1 as isolated domains are bound by recombinant NCp8 protein with high affinity, contrary to the hairpins downstream of SL1. Foot-printing of the SL1/NCp8 complex indicates that the major binding site maps to the SL1 upper stem. Taken together, these data suggest a model in which TAR hairpin III, the segment of SL1 proximal to the loop and the PAL palindromic sequence play specific roles in the initiation of dimerization. PMID- 21622660 TI - PHUSER (Primer Help for USER): a novel tool for USER fusion primer design. AB - Uracil-Specific Exision Reagent (USER) fusion is a recently developed technique that allows for assembly of multiple DNA fragments in a few simple steps. However, designing primers for USER fusion is both tedious and time consuming. Here, we present the Primer Help for USER (PHUSER) software, a novel tool for designing primers specifically for USER fusion and USER cloning applications. We also present proof-of-concept experimental validation of its functionality. PHUSER offers quick and easy design of PCR optimized primers ensuring directionally correct fusion of fragments into a plasmid containing a customizable USER cassette. Designing primers using PHUSER ensures that the primers have similar annealing temperature (T(m)), which is essential for efficient PCR. PHUSER also avoids identical overhangs, thereby ensuring correct order of assembly of DNA fragments. All possible primers are individually analysed in terms of GC content, presence of GC clamp at 3'-end, the risk of primer dimer formation, the risk of intra-primer complementarity (secondary structures) and the presence of polyN stretches. Furthermore, PHUSER offers the option to insert linkers between DNA fragments, as well as highly flexible cassette options. PHUSER is publicly available at http://www.cbs.dtu.dk/services/phuser/. PMID- 21622661 TI - Conservation of transcription factor binding events predicts gene expression across species. AB - Recent technological advances have made it possible to determine the genome-wide binding sites of transcription factors (TFs). Comparisons across species have suggested a relatively low degree of evolutionary conservation of experimentally defined TF binding events (TFBEs). Using binding data for six different TFs in hepatocytes and embryonic stem cells from human and mouse, we demonstrate that evolutionary conservation of TFBEs within orthologous proximal promoters is closely linked to function, defined as expression of the target genes. We show that (i) there is a significantly higher degree of conservation of TFBEs when the target gene is expressed in both species; (ii) there is increased conservation of binding events for groups of TFs compared to individual TFs; and (iii) conserved TFBEs have a greater impact on the expression of their target genes than non conserved ones. These results link conservation of structural elements (TFBEs) to conservation of function (gene expression) and suggest a higher degree of functional conservation than implied by previous studies. PMID- 21622662 TI - TimeTree2: species divergence times on the iPhone. AB - SUMMARY: Scientists, educators and the general public often need to know times of divergence between species. But they rarely can locate that information because it is buried in the scientific literature, usually in a format that is inaccessible to text search engines. We have developed a public knowledgebase that enables data-driven access to the collection of peer-reviewed publications in molecular evolution and phylogenetics that have reported estimates of time of divergence between species. Users can query the TimeTree resource by providing two names of organisms (common or scientific) that can correspond to species or groups of species. The current TimeTree web resource (TimeTree2) contains timetrees reported from molecular clock analyses in 910 published studies and 17 341 species that span the diversity of life. TimeTree2 interprets complex and hierarchical data from these studies for each user query, which can be launched using an iPhone application, in addition to the website. Published time estimates are now readily accessible to the scientific community, K-12 and college educators, and the general public, without requiring knowledge of evolutionary nomenclature. AVAILABILITY: TimeTree2 is accessible from the URL http://www.timetree.org, with an iPhone app available from iTunes (http://itunes.apple.com/us/app/timetree/id372842500?mt=8) and a YouTube tutorial (http://www.youtube.com/watch?v=CxmshZQciwo). PMID- 21622663 TI - Computational discovery of human coding and non-coding transcripts with conserved splice sites. AB - MOTIVATION: Long non-coding RNAs (lncRNAs) resemble protein-coding mRNAs but do not encode proteins. Most lncRNAs are under lower sequence constraints than protein-coding genes and lack conserved secondary structures, making it hard to predict them computationally. RESULTS: We introduce an approach to predict spliced lncRNAs in vertebrate genomes combining comparative genomics and machine learning. It is based on detecting signatures of characteristic splice site evolution in vertebrate whole genome alignments. First, we predict individual splice sites, then assemble compatible sites into exon candidates, and finally predict multi-exon transcripts. Using a novel method to evaluate typical splice site substitution patterns that explicitly takes the species phylogeny into account, we show that individual splice sites can be accurately predicted. Since our approach relies only on predicted splice sites, it can uncover both coding and non-coding exons. We show that our predicted exons and partial transcripts are mostly non-coding and lack conserved secondary structures. These exons are of particular interest, since existing computational approaches cannot detect them. Transcriptome sequencing data indicate tissue-specific expression patterns of predicted exons and there is evidence that increasing sequencing depth and breadth will validate additional predictions. We also found a significant enrichment of predicted exons that form multi-exon transcript parts, and we experimentally validate such a novel multi-exon gene. Overall, we obtain 336 novel multi-exon transcript predictions from human intergenic regions. Our results indicate the existence of novel human transcripts that are conserved in evolution and our approach contributes to the completion of the human transcript catalog. AVAILABILITY AND IMPLEMENTATION: Predicted human splice sites, exons and gene structures together with a Perl implementation of the tree-based log-odds scoring and a supplementary PDF file containing additional figures and tables are available at: http://www.bioinf.uni-leipzig.de/publications/supplements/10-010. The five experimentally confirmed partial transcript isoforms have been deposited in GenBank under accession numbers HM587422-HM587426. PMID- 21622664 TI - RightField: embedding ontology annotation in spreadsheets. AB - MOTIVATION: In the Life Sciences, guidelines, checklists and ontologies describing what metadata is required for the interpretation and reuse of experimental data are emerging. Data producers, however, may have little experience in the use of such standards and require tools to support this form of data annotation. RESULTS: RightField is an open source application that provides a mechanism for embedding ontology annotation support for Life Science data in Excel spreadsheets. Individual cells, columns or rows can be restricted to particular ranges of allowed classes or instances from chosen ontologies. The RightField-enabled spreadsheet presents selected ontology terms to the users as a simple drop-down list, enabling scientists to consistently annotate their data. The result is 'semantic annotation by stealth', with an annotation process that is less error-prone, more efficient, and more consistent with community standards. AVAILABILITY AND IMPLEMENTATION: RightField is open source under a BSD license and freely available from http://www.rightfield.org.uk PMID- 21622665 TI - The biotron breeding system: a rapid and reliable procedure for genetic studies and breeding in rice. AB - Oryza sativa is widely used as a model organism for many aspects of research in monocots and cereals. However, it has certain disadvantages as a model species compared with Arabidopsis thaliana, the eudicot species most widely used in plant sciences: first, it has a long cultivation time; and second, it requires considerably more space for growth. Here, we introduce a biotron breeding system, which allows rapid and reliable rice cultivation using a well-equipped artificial environmental chamber. This system involves use of regulation of CO2 levels, removal of tillers and embryo rescue to overcome the disadvantages of rice cultivation. The rice cultivars Nipponbare, Koshihikari, Taichung 65 and Kasalath all showed vigorous growth and sufficient seed production in the biotron breeding system with accelerated flowering time. Nipponbare, which was the earliest among these cultivars, flowered at about 50 d after sowing. The life cycle of these plants could be further shortened using an embryo rescue technique on immature seeds at 7 d after pollination, thereby avoiding the lengthy process of seed maturation. Overall, it was possible to shorten the life cycle of Nipponbare to about 2 months under the controlled conditions. Furthermore, controlled crosses, which can be difficult with conventional cultivation methods, were easy to perform as we could control the exact timing of anther dehiscence. Thus, our biotron breeding system offers a valuable new approach to genetic and breeding studies in rice. PMID- 21622666 TI - Bibliometrics of anaesthesia researchers in the UK. AB - BACKGROUND: Bibliometrics provide surrogate measures of the quality and quantity of research undertaken by departments and individuals. Previous reports have suggested that academic anaesthesia research in the UK is in decline. We wished to provide a comprehensive description of current and historical published output of UK anaesthesia researchers. METHODS: Bibliometric indices (Web of Science((r))) were calculated for anaesthesia researchers in the UK for the whole period covered by the database, and for 2004-8. A parallel search was made using the ScholarometerTM tool, which parses output from Google ScholarTM. Calculated indices included total number of publications; total number of citations; citations per paper; h-index; g-index; and modified impact index. RESULTS: One hundred and four individuals and 23 academic departments were identified. Median values (inter-quartile range) for the indices were: total papers 57 (24-95) (individuals for the whole period), 11 (6-20) (individuals 2004-8), 50 (30-70) (departments 2004-8); total number of citations 571 (175-1328), 93 (38-207), 383 (239-845); h-index 13 (8-20), 6 (3-8), 11 (9-14). Four departments were ranked in the top 5 for all indices. CONCLUSIONS: The general distribution of bibliometric data is similar to that seen in other specialities in Europe and North America. Four departments contribute to more than 50% of published anaesthesia research output in this data set. These data provide useful comparative tools for individuals, departments, and national bodies. PMID- 21622667 TI - Cell-free RNA replication systems based on a human cell extracts-derived in vitro translation system with the encephalomyocarditisvirus RNA. AB - To study the relationship between translation and replication of encephalomyocarditisvirus (EMCV) RNA, we established a cell-free RNA replication system by employing a human cell extracts-based in vitro translation system. In this system, a cis-EMCV RNA replicon encoding the Renilla luciferase (R-luc) or GFP and the viral regulatory proteins efficiently replicated with simultaneous translation of the encoded protein. To examine how translation of the replicon RNA, but not the translated products, affected replication, a trans-EMCV RNA replicon encoding R-luc and the RNA replication elements was next constructed. The trans-replicon RNA replicated only in the presence of the regulatory proteins pre-expressed in trans. Incubation with cycloheximide, puromycin or a dominant negative eukaryotic translation initiation factor 4A following expression of the regulatory proteins almost completely inhibited not only translation of the trans replicon RNA but also replication of the RNA, suggesting that EMCV RNA translation promotes replication of the RNA. In conclusion, the cell-free RNA replication systems should become useful tools for the study of the viral RNA replication. PMID- 21622668 TI - Living on an atrial kick--an unusual case of a stuck mitral valve. PMID- 21622669 TI - Sirolimus-eluting stents vs. bare-metal stents for treatment of focal lesions in infrapopliteal arteries: a double-blind, multi-centre, randomized clinical trial. AB - AIMS: Preliminary reports indicate that sirolimus-eluting stents reduce the risk of restenosis after percutaneous infrapopliteal artery revascularization. We conducted a prospective, randomized, multi-centre, double-blind trial comparing a polymer-free sirolimus-eluting stent with a placebo-coated bare-metal stent in patients with either intermittent claudication or critical limb ischaemia who had a de-novo lesion in an infrapopliteal artery. METHODS AND RESULTS: 161 patients were included in this trial. The mean target lesion length was 31 +/- 9 mm. The main study endpoint was the 1-year primary patency rate, defined as freedom from in-stent-restenosis (luminal narrowing of >=50%) detected with duplex ultrasound if not appropriate with angiography. Secondary endpoints included the 6-month primary patency rate, secondary patency rate, and changes in Rutherford-Becker classification after 1 year. Twenty-five (15.5%) patients died during the follow up period. One hundred and twenty-five patients reached the 1-year examinations. The 1-year primary patency rate was significantly higher in the sirolimus-eluting stent group (80.6%) than in the bare-metal stent group (55.6%, P= 0.004), and the 1-year secondary patency rates were 91.9 and 71.4% (P= 0.005), respectively. The median (interquartile range) change in Rutherford-Becker classification after 1 year was -2 (-3 to -1) in the sirolimus-eluting stent group and -1 (-2 to 0) in the bare-metal stent group, respectively (P= 0.004). CONCLUSION: Mid-term patency rates of focal infrapopliteal lesions are substantially improved with sirolimus eluting stent compared with bare-metal stent. Corresponding to the technical results, the changes in Rutherford-Becker classification reveal a significant advantage for the sirolimus-eluting stent. PMID- 21622670 TI - EMLA versus glucose for PICC insertion: a randomised triple-masked controlled study. PMID- 21622671 TI - Scalp laceration in a newborn due to cervical sutures. PMID- 21622672 TI - Factors related to care home admission in the year following hospitalisation in frail older adults. PMID- 21622673 TI - Telomere length and anaemia in old age: results from the Newcastle 85-plus Study and the Leiden 85-plus Study. AB - BACKGROUND: reduced telomere length in blood cells has been associated with increased risk of multiple age-related diseases and is widely regarded as a general biomarker of ageing. Therefore, it is important to know both the extent and limitations of this association. We investigated the relation between telomere length and anaemia in two independent cohorts, with the prior expectation of adding anaemia to the list of conditions for which telomere reduction is a risk factor. PARTICIPANTS AND METHODS: the present study is embedded in the Newcastle 85-plus Study and Leiden 85-plus Study, two population based studies of inhabitants of Newcastle and North Tyneside, UK (n = 749) and Leiden, the Netherlands (n = 658) aged 85 and over. High-molecular-weight DNA was isolated from full fresh blood (Newcastle) and peripheral blood mononuclear cells samples (Leiden). Telomere length was measured as abundance of telomeric template versus a single gene by quantitative real-time polymerase chain reaction. Anaemia was defined according to World Health Organization criteria. RESULTS: in both studies, no differences in median telomere length were observed between participants with anaemia and participants without anaemia (Newcastle: 2,846 bp (interquartile range (IQR) 2,433-3,630) versus 2,920 bp (IQR 2,425-3,570), P = 0.63; Leiden: 4,136 bp (IQR 3,879-4,428) versus 4,167 bp (IQR 3,893-4,501), P = 0.41). Telomere length also did not correlate with any other haematological parameter in both men and women. CONCLUSIONS: in contrast to other age-related diseases, telomere length is not associated with anaemia or any other haematological parameter in older individuals in the general population. PMID- 21622674 TI - European Surveillance of Antimicrobial Consumption (ESAC): systemic antiviral use in Europe. AB - OBJECTIVES: To assess the total systemic antiviral use in Europe and to identify the antiviral substances most commonly used. METHODS: Within the European Surveillance of Antimicrobial Consumption (ESAC; www.esac.ua.ac.be), using the anatomical therapeutic chemical (ATC) classification and defined daily dose (DDD) measurement unit, data on total (out- and inpatient) systemic antiviral use (ATC J05), aggregated at the level of the active substance, were collected for 2008, and use was expressed in DDD (WHO ATC/DDD, version 2010) per 1000 inhabitants per day (DID). Antiviral substances were grouped according to their main indication. RESULTS: In Europe, 12 countries (Belgium, Croatia, Denmark, Estonia, Finland, France, Hungary, Italy, Luxembourg, Russia, Slovenia and Sweden) provided total (out- and inpatient) data and 4 countries (Austria, the Netherlands, Portugal and Norway) provided outpatient data only. Total systemic antiviral use varied by a factor of 10.95 between the country with the highest (3.53 DID in France) and the country with the lowest (0.32 DID in Croatia) use. HIV/AIDS antivirals represented more than 50% of the total antiviral use in most countries. The amount and spectrum of antivirals used varied greatly between countries. CONCLUSIONS: Our study demonstrated a wide variation of total systemic antiviral use in several European countries, as striking as that of outpatient systemic antibiotic, antimycotic and antifungal use. The variation is mainly determined by the use of HIV/AIDS antivirals. These observations should stimulate further analysis to understand the variation of specific antiviral substances. The ESAC data facilitate auditing of antiviral prescriptions and evaluation of the implementation of guidelines and public health policies. PMID- 21622675 TI - Too much and too little? Prevalence and extent of antibiotic use in a New Zealand region. AB - OBJECTIVES: Although antibiotic use in the community is a significant contributor to resistance, little is known about social patterns of use. This study aimed to explore the use of antibiotics by age, gender, ethnicity, socio-economic status and rurality. METHODS: Data were obtained on all medicines dispensed to ambulatory patients in one isolated town for a year, and data on antibiotics are presented in this paper. Demographic details were obtained from pharmacy records or by matching to a national patient dataset. RESULTS: During the study year, 51% of the population received a prescription for one or more antibiotics, and on average people in the region received 10.15 defined daily doses (DDDs). Prevalence of use was higher for females (ratio, 1.18), and for young people (under 25) and the elderly (75 and over), and the amount in DDDs/person/year broadly followed this pattern. Maori (indigenous New Zealanders) were less likely to receive a prescription (48% of the population) than non-Maori (55%) and received smaller quantities on average. Rural Maori, including rural Maori children, received few prescriptions and low quantities of antibiotics compared with other population groups. CONCLUSIONS: The level of antibiotic use in the general population is high, despite campaigns to try to reduce unnecessary use. The prevalence of acute rheumatic fever is high amongst rural Maori, and consequently treatment guidelines recommend prophylactic use of antibiotics for sore throat in this population. This makes the comparatively very low level of use of antibiotics amongst rural Maori children very concerning. PMID- 21622676 TI - A randomized clinical trial of a Therapeutic Workplace for chronically unemployed, homeless, alcohol-dependent adults. AB - AIMS: To assess the efficacy of the Therapeutic Workplace, a substance abuse intervention that promotes abstinence while simultaneously addressing the issues of poverty and lack of job skills, in promoting abstinence from alcohol among homeless alcoholics. METHODS: Participants (n = 124) were randomly assigned to conditions either requiring abstinence from alcohol to engage in paid job skills training (Contingent Paid Training group), offering paid job skills training with no abstinence contingencies (Paid Training group) or offering unpaid job skill training with no abstinence contingencies (Unpaid Training group). RESULTS: Participants in the Contingent Paid Training group had significantly fewer positive (blood alcohol level >= 0.004 g/dl) breath samples than the Paid Training group in both randomly scheduled breath samples collected in the community and breath samples collected during monthly assessments. The breath sample results from the Unpaid Training group were similar in absolute terms to the Contingent Paid Training group, which may have been influenced by a lower breath sample collection rate in this group and fewer reported drinks per day consumed at intake. CONCLUSION: Overall, the results support the utility of the Therapeutic Workplace intervention to promote abstinence from alcohol among homeless alcoholics, and support paid training as a way of increasing engagement in training programs. PMID- 21622677 TI - Socioeconomic position and effectiveness of lifestyle intervention in prevention of type 2 diabetes: one-year follow-up of the FIN-D2D project. AB - AIMS: Lifestyle intervention is effective in prevention of type 2 diabetes (T2D) in high-risk individuals. However, health behaviour and health outcomes are modified by socioeconomic position through various mechanisms. It is therefore possible that success in lifestyle intervention may be determined by factors such as level of education or occupation. In this study we assessed the impact of the level of education and occupation on the baseline anthropometric and clinical characteristics and their changes during a one-year follow-up in a cohort of Finnish men and women at high risk for T2D aged 20-64 years. METHODS: As part of a Finnish national diabetes prevention programme 2003-2007 (FIN-D2D), high-risk individuals were identified using opportunistic screening for lifestyle intervention in primary health care. 1,067 men and 2,122 women had one-year follow-up data. Education and occupation were used as factors of socioeconomic position. Measures of anthropometric and clinical characteristics included weight, body mass index (BMI), waist circumference, systolic and diastolic blood pressure, total, HDL and LDL cholesterol, triglycerides, FINDRISC scores and glucose tolerance status. RESULTS: The effect of intervention was similar in all socioeconomic groups, but the level of education was related to glucose tolerance status in both genders. In addition, socioeconomic differences existed in blood pressure, weight, BMI, waist circumference and HDL cholesterol. CONCLUSIONS: Socioeconomic position did not seem to have any impact on the effectiveness of lifestyle intervention in individuals at high risk for T2D, which is encouraging from the point of view of reducing health inequalities. PMID- 21622678 TI - Abstinence-orientated residential rehabilitation of opioid users in Denmark: do changes in national treatment policies affect treatment outcome? AB - AIMS: Significant changes in the Danish drug-abuse treatment system occurred from 1998 to 2008, allowing the opportunity to study their impact on outcomes for opiate dependent patients. This paper examines whether such changes are related to possible changes in abstinence rates of two cohorts of drug users. METHODS: We compared survival curves and the month-by-month probability of abstinence between cohorts of drug abusers who were followed for one year after treatment for opioid dependence in 1998 (n = 305) and in 2008 (n = 204). Of the 1998 cohort, 192 were interviewed face-to-face and 61 were found through national drug-abuse treatment registers. Of the 2008 cohort 112 were interviewed face-to-face and 42 were found through national drug-abuse treatment registers. RESULTS: Survival curves did not differ between the two cohorts. After controlling for age and gender, people from the 2008 cohort had a higher frequency of month-to-month rates of abstinence. A higher proportion of clients received new treatment after their index treatment in 2008. Clients from the 1998 cohort who did not receive new treatment used drugs more often than clients who did not receive treatment in 2008. At the same time, the proportion of clients who received residential rehabilitation treatment for opiate dependence increased substantially by 2008. CONCLUSIONS: Total abstinence rates are highly consistent over time and seem to change little with changes in systems of care. However, changes in care that improve access to treatment may reduce the overall burden of opioid addiction to both individuals and society. PMID- 21622679 TI - In vivo distribution of lead in male and female rats after intraperitoneal and oral administration. AB - The resultant effects of lead exposure are seen in almost all the systems of the body and results in toxicity to many organs. Since toxicity depends on its degree of uptake, distribution and metabolism, the authors investigated the differential uptake, accumulation and distribution of lead in organs of males and female Wistar rats following various routes of administration. Group 1 served as control male and control female; group 2 males and females received 5 mg/kg body weight of lead intraperitoneally for 8 days while group 3 males and female rats were administered drinking water containing 100 ppm of lead acetate for 18 days. Tissues were collected for analysis of the lead content using atomic absorption spectroscopy. The relative retention of lead by the tissues was greater in rats exposed to lead by the i.p. route varying in the order of accumulation / uptake in males as lungs > spleen > stomach > kidney > blood > heart and in females as spleen > stomach > heart > kidney > blood > lungs (i.p. route) and (oral route) as for males kidney > lungs > stomach > blood > heart > spleen, and females as kidney > lungs > stomach > blood > heart > spleen. Male Wistar rats showed more accumulation with oral exposure in lungs, spleen and blood with values for kidney and stomach being significantly (p < 0.05) higher when compared with females. Female Wistar rats showed more accumulation with i.p. exposure for spleen and stomach tissues while values for the heart was significantly (p < 0.05) higher than the males. Our findings suggest that lead retention and the organ distribution varied depending upon the sex and route of lead administration. PMID- 21622680 TI - MicroRNA-195 promotes palmitate-induced apoptosis in cardiomyocytes by down regulating Sirt1. AB - AIMS: Free fatty acids induce apoptosis in cardiomyocytes, which is implicated in lipotoxic cardiomyopathy. However, the underlying mechanisms remain not fully understood. MicroRNAs (miRNAs) are non-coding small RNAs that control gene expression at the post-transcriptional level. Dysregulated miRNAs have been shown to be involved in heart diseases. This study was to examine whether miR-195 regulates palmitate-induced cardiomyocyte apoptosis by targeting Sirt1, a known anti-apoptotic protein. METHODS AND RESULTS: In cultured neonatal mouse cardiomyocytes, palmitate up-regulated miR-195 expression, increased reactive oxygen species (ROS) production, and induced apoptosis as determined by up regulation of caspase-3 activity and DNA fragmentation. Inhibition of miR-195 decreased ROS production and apoptosis in palmitate-stimulated cardiomyocytes. In contrast, a miR-195 mimic enhanced palmitate-induced ROS production and apoptosis. The induction of miR-195 correlated with a reduction in Sirt1 and Bcl 2. We further showed that miR-195 targeted and inhibited Sirt1 expression through two target sites located in the 3' un-translational region of Sirt1 mRNA. In concordance, inhibition of miR-195 increased Sirt1 protein in cardiomyocytes whereas the miR-195 mimic reduced it. Activation of Sirt1 or overexpression of Bcl-2 inhibited palmitate-induced apoptosis. On the other hand, inhibition of Sirt1 enhanced apoptosis. The inhibitory effect of Sirt1 on apoptosis was associated with a reduction in ROS. CONCLUSIONS: This study demonstrates a pro apoptotic role of miR-195 in cardiomyocytes and identifies Sirt1 as a direct target of miR-195. The effect of miR-195 on apoptosis is mediated through down regulation of Sirt1 and Bcl-2 and ROS production. Thus, miR-195 may be a new therapeutic target for lipotoxic cardiomyopathy. PMID- 21622681 TI - IL-6-mediated Th17 differentiation through RORgammat is essential for the initiation of experimental autoimmune myocarditis. AB - AIMS: Interleukin (IL)-17-producing helper T (Th17) cells have been proposed to participate in the pathogenesis of chronic inflammation, such as autoimmune myocarditis. IL-6 gene ablation confers the resistance to experimental autoimmune myocarditis (EAM). In this study, we have addressed the pathological roles of IL 6 in the regulation of Th17 cells in EAM. METHODS AND RESULTS: To induce EAM, mice were immunized twice with alpha-myosin heavy chain peptide. Three weeks after the first injection, the cardiac expression of the Th17-specific transcription factor, retinoic acid receptor-related orphan nuclear receptor (ROR gammat), was up-regulated. Consistently, Th17 cells were recruited into EAM hearts, as analysed by flow cytometry. Using the mice with enhanced green fluorescence protein (eGFP) gene knocked-in at RORgammat locus (RORgammat-eGFP mice), we observed Th17 cell infiltration into inflamed lesions. Pre-treatment with IL-6 receptor (IL-6R)-blocking antibody (anti-IL-6R Ab) inhibited EAM induction in terms of disease severity score (3.5 +/- 0.8; IgG vs. 0.5 +/- 0.8; anti-IL-6R Ab, n = 6, P< 0.01) and suppressed the myocardial expression of IL-17 and RORgammat. In contrast, the administration of anti-IL-6R Ab 7 days after the first immunization failed to show the inhibitory effects, suggesting that IL-6 plays important roles in EAM initiation. Finally, by generating RORgammat-eGFP homozygous mice, we revealed that RORgammat gene ablation conferred the resistance to EAM induction. CONCLUSION: IL-6-mediated induction of Th17 cells is critical for the onset of EAM, but not for its progression. IL-6/Th17 signalling could be a promising therapeutic target for the prevention of myocardial inflammation. PMID- 21622682 TI - Contributions of ACE and mast cell chymase to endogenous angiotensin II generation and leucocyte recruitment in vivo. AB - AIMS: In vitro studies suggest that mast cell chymase (MCP) is more important than angiotensin-converting enzyme (ACE) for generating angiotensin II (Ang II) within the cardiovascular system. We investigated in vivo the relative contributions of ACE and MCP to leucocyte recruitment induced by endogenously generated Ang II. METHODS AND RESULTS: Exposure of the murine cremasteric microcirculation of C57BL/6 mice to Ang I (100 nM for 4 h) induced leucocyte endothelium interactions. Either losartan (an Ang II receptor-1 antagonist, AT(1)) or enalapril (an ACE inhibitor), but not chymostatin (a chymase inhibitor), inhibited Ang I-induced responses. Mast cell degranulation with compound 48/80 (CMP48/80, 1 MUg/mL) also induced leucocyte adhesion but this was only weakly affected by the inhibitors. When Ang I and CMP48/80 were co administered, AT(1B) receptor expression was increased, MCP-4 was found surrounding the vessel wall, and ACE was detected in the endothelium. Ang I + CMP48/80 induced enhanced leucocyte adhesion that was attenuated by losartan, enalapril, enalapril + chymostatin, and cromolyn (a mast cell stabilizer). The use of male mast cell-deficient WBB6F1/J-Kit(w)/Kit(w-v) mice (C57BL/6 background) confirmed these findings. CONCLUSION: In vivo, Ang II is primarily generated by ACE under basal conditions, but in inflammatory conditions, the release of MCP amplifies local Ang II concentrations and the associated inflammatory process. Thus, AT(1) receptor antagonists may be more effective than ACE inhibitors for treating ongoing Ang II-mediated vascular inflammation. PMID- 21622683 TI - Role of T-type calcium channel subunits in post-myocardial infarction remodelling probed with genetically engineered mice. AB - AIMS: Previous studies suggested that T-type Ca(2+)-current (I(CaT))-blockers improve cardiac remodelling, but all available I(CaT)-blockers have non-specific actions on other currents and/or functions. To clarify the role of I(CaT) in cardiac remodelling, we studied mice with either of the principal cardiac I(CaT) subunits (Cav3.1 or Cav3.2) knocked out. METHODS AND RESULTS: Adult male Cav3.1- or Cav3.2-knockout (Cav3.1(-/-), Cav3.2(-/-)) mice and respective wild-type (WT) littermate controls were subjected to left anterior descending coronary artery ligation to create myocardial infarction (MI). Echocardiography and programmed electrical stimulation were performed at baseline and 4 weeks post-MI. At baseline, Cav3.1(-/-) mice had slowed heart rates and longer PR intervals vs. WT, but no other electrophysiological and no haemodynamic differences. Cav3.2(-/-) showed no differences vs. WT. Contractile indices (left ventricular fractional shortening and ejection fraction) decreased more post-MI in Cav3.1(-/-) mice than in Cav3.1(+/+) (e.g. by 34 and 29% for WT; 50 and 45% for Cav3.1(-/-), respectively; P < 0.05 for each). Cav3.1(-/-) mice had increased ventricular tachycardia (VT) inducibility post-MI (9 of 11, 82%) vs. WT (3 of 10, 30%; P < 0.05). Cav3.2(-/-) mice were not different in cardiac function or VT inducibility vs. WT. Quantitative polymerase chain reaction showed that Cav3.1 is the major I(CaT)-subunit and that no compensatory Cav3.2 up-regulation occurs in Cav3.1(-/ ) mice. Cav3.1(-/-) and Cav3.2(-/-) mice had no mRNA expression for the knocked out gene, at baseline or post-MI. CONCLUSION: Our findings suggest that, contrary to suggestions from previous studies with (imperfectly selective) pharmacological agents having T-type Ca(2+)-channel-blocking actions, elimination of Cav3.1 expression leads to impaired cardiac function and enhanced arrhythmia vulnerability post-MI, whereas Cav3.2 elimination has no effect. PMID- 21622684 TI - Morning glory disc anomaly and optic nerve coloboma. PMID- 21622685 TI - Recurrent group B streptococcal bacteremia. PMID- 21622686 TI - Age and disability biases in pediatric resuscitation among future physicians. AB - This study examined whether biases concerning age and/or disability status influenced resuscitation decisions. Medical students were randomly chosen to read 1 of 4 vignettes, organized in a 2 (age: infant vs school-age) * 2 (disability: preexisting vs no preexisting) between-subjects design. The vignettes described a pediatric patient experiencing an acute episode who required resuscitation. Following resuscitation, patients with existing disability would continue to have disability, whereas those without would develop disability. Participants indicated whether they would resuscitate, given a 10% chance of success. There was a significant main effect of disability: Medical students displayed a preference for resuscitating previously disabled children compared with previously healthy children when prognosis was held constant, F(1, 121) = 4.89, p = .03. This differential treatment of the two groups cannot easily be morally justified and poses a quandary for educators. PMID- 21622687 TI - Scrotal hair in the first year of life. PMID- 21622688 TI - Severe anemia and seborrhea in an 18-month-old child. PMID- 21622689 TI - Short-dwell ethanol lock therapy in children is associated with increased clearance of central line-associated bloodstream infections. AB - BACKGROUND: Central line-associated bloodstream infection (CLABSI) is a known complication of central line use. Salvage of infected central lines with ethanol lock therapy (ELT) with systemic antimicrobials may be an alternative treatment option in children. METHODS: Retrospective review was performed in children with CLASBI who underwent short-dwell ELT (70% ethanol, 4- to 25-hour dwell times <=3 days) with systemic antimicrobials from January 1, 2007 to July 15, 2009. RESULTS: A total of 59 patients, aged 2 months to 19 years (mean +/- SD = 6.3 +/- 6.1 years) with 80 episodes of CLABSI were included. The CLABSI eradication rate was 86% (69/80 episodes; 95% confidence interval [CI] 78%, 94%), significantly greater than 50% (Z = 2.35, P < .05), the estimated clearance rate of CLABSI eradication using systemic antimicrobials alone. Overall central line retention was 78% (60/77 episodes, 95% CI 69%, 87%). ELT was well tolerated. CONCLUSIONS: These findings suggest the potential benefit of short-dwell ELT combined with systemic antimicrobials in CLABSI treatment. Randomized controlled trials are needed. PMID- 21622690 TI - Appointment attendance in a pediatric weight management clinic. PMID- 21622691 TI - Incidence of traumatic lumbar puncture: experience of a large, tertiary care pediatric hospital. AB - The objective of this study was to establish the incidence of traumatic or unsuccessful lumbar punctures (LPs) in the authors' institution. This is a prospective study. Traumatic LP was defined as >400 red blood cells (RBCs) and unsuccessful LP as failure to obtain cerebrospinal fluid (CSF) after the first LP attempt. A total of 127 CSF were recorded over 1 year. The incidence of a traumatic LP was 27/103 (26.2%) using the sitting position and 3/24 (12.5%) in infants and children using the lying position. In total, 33 (26%) CSF samples had 0 RBCs. The incidence of traumatic LP was 24% when the procedure involved one LP and 50% when more than one attempt was required. RBC count was significantly lower in cases requiring only one attempt (P = .0074). Incidence of traumatic LP was independent of physicians' experience, sedation use or time of procedure, suggesting an intrinsic factor as the cause of traumatic LPs. PMID- 21622692 TI - Anticipatory guidance and early child development: pediatrician advice, parent behaviors, and unmet needs as reported by parents from different backgrounds. AB - Anticipatory guidance is acknowledged as an important aspect of pediatric practice, and research shows that parents expect and value the information and advice they receive from their pediatric health care providers. This study surveyed 373 parents (88% mothers) of infants in waiting rooms of diverse pediatric practices in Memphis,Tennessee, about their recall of anticipatory guidance received, unmet needs for anticipatory guidance, preferences for sources of information, and activities to promote healthy development. Most parents recalled specific guidance received (diet, communication, and discipline being the most common), and unmet needs (discipline and development being among the most common). Disadvantaged parents reported the fewest unmet needs. Most parents readily named specific activities they engage in to promote development, including reading, educational toys, and nurturing. PMID- 21622693 TI - Repeated ovarian stimulation with corifollitropin alfa in patients in a GnRH antagonist protocol: no concern for immunogenicity. AB - BACKGROUND: One injection of corifollitropin alfa replaces the first seven daily FSH injections in controlled ovarian stimulation (COS) cycles. Repeated treatment with therapeutic proteins may cause immune responses or hypersensitivity reactions. We assessed the immunogenicity and safety of corifollitropin alfa treatment in up to three COS cycles. METHODS: In this multicentre, phase III uncontrolled trial, patients (>60 kg) started treatment with one injection of 150 ug corifollitropin alfa on cycle Day 2 or 3 of menses and 0.25 mg ganielix on stimulation Day 5 or 6. Primary outcome measures were antibody formation against corifollitropin alfa (using highly sensitive radioimmunoprecipitation assay), hypersensitivity reactions, local tolerance and adverse events (AEs). RESULTS: First, second and third COS cycles were started by 682, 375 and 198 patients, respectively. No clinically relevant immunogenicity or drug-related hypersensitivity was observed. For 192 patients undergoing their third cycle a post-treatment blood sample was negative in the anti-corifollitropin antibody assay, resulting in an upper limit of the one-sided 95% confidence interval (CI) of 1.5%. Most frequent AEs were procedural pain (17.7%, 95% CI: 14.9-20.8%), headache (9.1%, 95% CI: 7.0-11.5%) and pelvic pain (7.6%, 95% CI: 5.7-9.9%). Cumulative ongoing pregnancy rate after three cycles, including frozen-thawed embryo transfer cycles and spontaneous pregnancies, was 61% (95% CI: 56-65%) after censoring for patients who discontinued. CONCLUSIONS: Treatment with corifollitropin alfa can safely and effectively initiate and sustain ovarian stimulation during the first 7 days of COS in normal responder patients undergoing up to three treatment cycles, without concerns of immunogenicity. The trial was registered under ClinicalTrials.gov identifier NCT00696878. PMID- 21622694 TI - Fukushima Daiichi and Icarus: the human factor in a meltdown (Sv=1J/kg.w). PMID- 21622695 TI - The anglerfish and uremic toxins. PMID- 21622696 TI - The power of Pasteur's quadrant: cardiovascular disease at the turn of the century. AB - During the life span of The FASEB Journal, the decline in cardiovascular mortality was astonishing as the fundamental bases of the complex syndromes of cardiovascular disease were illuminated. In this Silver Anniversary Review, we highlight a few pivotal advances in the field and relate them to research in Pasteur's quadrant, the region of investigation driven by both a desire for fundamental understanding and the consideration of its use. In the second half of the 20th century, we advanced from little pathophysiologic understanding to a near-complete understanding and effective, evidence-based therapeutics for vascular disorders and a similar development of pharmacotherapy to address heart failure, primarily through agents that antagonize the excessive concentration of circulating neurohumoral agents. In the current era, we have witnessed "the rise of the machines," from stents to cardiac resynchronization therapy. The next wave of treatments will build on an increasingly sophisticated understanding of the molecular determinants of cardiovascular disorders. We briefly consider the promise of regenerative medicine and are intrigued by the possibility for the direct reprogramming of resident cardiac fibroblasts into cardiomyocytes. As for the future, genomic profiling should help physicians recommend individualized risk factor modification targeted to prevent specific manifestations of cardiovascular disease. Transcriptional and biomarker analyses will almost surely be used individually to tailor therapy for those at risk of or experiencing cardiovascular disease. Given the ongoing exponential expansion of scientific knowledge, all of human ingenuity will be needed to fully utilize the power of Pasteur's quadrant and to unleash another quarter century in cardiology as scientifically fruitful and effective on human health as the last. PMID- 21622700 TI - Endovascular coiling of the ophthalmic artery in pigs to induce retinal ischemia. AB - PURPOSE: The authors recently showed that the retinal circulation can be accessed by transfemoral endovascular catheterization. The purpose of this study was to examine whether endovascular coiling can be used to induce different degrees of ischemic injury. The possibility of creating occlusions at different sites in the vasculature to cause retinal ischemia with different degrees of severity was investigated. METHODS: The ophthalmic artery was catheterized through the external carotid system using a fluoroscopy-monitored, transfemoral, endovascular approach in 12 pigs (mean weight, 70 kg). The effects were evaluated using angiography and multifocal electroretinography. RESULTS: Occlusion of arteries supplying the retina was established using endovascular coiling. Coiling in the proximal part of the ophthalmic artery caused no or little ischemia, presumably because of collateral blood supply. Coiling in the distal part of the ophthalmic artery, over the branching of the main ciliary artery, caused more severe retinal ischemia. Multifocal electroretinography recordings, which reflect retinal function in an area close to the visual streak, showed decreased amplitudes and increased implicit times after distal occlusion, but not after proximal occlusion of the ophthalmic artery. The responses were similar 1 hour and 72 hours after coiling, indicating that a permanent ischemic injury was established. CONCLUSIONS: The porcine ophthalmic artery can be occluded using an endovascular coiling technique. This provides an experimental animal model of retinal ischemia in which occlusion at different sites of the vasculature produces different degrees of severity of the ischemic damage. PMID- 21622702 TI - Structural correlation between the nerve fiber layer and retinal ganglion cell loss in mice with targeted disruption of the Brn3b gene. AB - PURPOSE: Mice with a targeted disruption of Brn3b (knockout Brn3b(-/-)) undergo the loss of a majority of retinal ganglion cells (RGCs) before birth. Spectral domain optical coherence tomography (SD-OCT) allows for the noninvasive examination of Brn3b(-/-) cellular loss in vivo. METHODS: The central retinas of Brn3b(-/-) and phenotypically wild-type (Brn3b(+/+) and Brn3b(+/-)) mice were imaged by SD-OCT. The combined nerve fiber layer (NFL) and inner plexiform layer (IPL) were manually segmented and thickness maps were generated. The results were confirmed by histologic and immunofluorescence cell counts of the RGC layer (RGCL) of the same retinas. RESULTS: The combined NFL and IPL of the Brn3b(-/-) retinas were significantly thinner, and the histologic cell counts significantly lower, than those of the phenotypically wild-type retinas (paired t-test; P < 0.01 and P < 0.01, respectively). The combined NFL and IPL thickness and the histologic cell count correlated highly (R(2) = 0.9612). Immunofluorescence staining revealed significant RGC-specific loss in Brn3b(-/-) retinas (paired t test; P < 0.01). The distribution of combined central NFL and IPL loss was not localized or sectorial. CONCLUSIONS: The strong correlation between the combined layer thickness and histologic cell counts validates manual OCT segmentation as a method of monitoring cell loss in the RGCL. A retinal thickness map assessed if combined NFL and IPL thickness loss in Brn3b(-/-) eyes was topographically specific. Generalized RGC and combined NFL and IPL loss was observed in the Brn3b(-/-) retinas, in contrast to topographically specific RGC loss observed in glaucomatous DBA2/J eyes. PMID- 21622703 TI - Imaging of forward light-scatter by opacified posterior capsules isolated from pseudophakic donor eyes. AB - PURPOSE: Posterior capsule opacification (PCO) degrades visual function by reducing visual acuity, but also by increasing intraocular light-scatter. An in vitro model was used to elucidate the effect of PCO-morphology on light-scatter and its functional aspect, as can be assessed with straylight measurement. METHODS: Forward PCO-scatter by opacified capsular bags was recorded with a goniometer and camera. The camera position mimicked the anatomic position of retinal photoreceptors; the camera recorded the scattered light that the photoreceptors would sense in an in vivo situation. Scattered light was recorded at different wavelengths and scatter angles, which were divided into a near (1 degrees < theta <= 7 degrees ) and far (theta > 7 degrees ) large-angle domain. Using scattered light, the camera produced grayscale PCO images. The nature of the angular dependence of PCO-scatter was compared with that of scatter in the normal eye, by rescaling PCO images relative to the normal eye's point-spread function. RESULTS: The scattered light images closely followed PCO severity. The angular dependence of PCO-scatter resembled that of scatter in the normal eye, irrespective of severity and PCO type. PCO shows the type of wavelength dependence that is normal for small particles: monotonically decreasing with increasing wavelength. At the near large-angle domain, the angular dependence of PCO scatter resembled the angular dependence of scatter in the normal eye less closely. CONCLUSIONS: Surprisingly, PCO scatter and scatter in the normal eye have similar underlying scattering processes. However, data obtained at the near large-angle domain demonstrates that, apart from scatter, PCO may also have a refractile component, which is most pronounced in pearl-type PCO. PMID- 21622704 TI - Macular choroidal thickness and volume in normal subjects measured by swept source optical coherence tomography. AB - PURPOSE: To study the choroidal thickness in healthy subjects by swept-source optical coherence tomography (SS-OCT) at longer wavelength. METHODS: The macular area of 31 eyes (31 healthy volunteers) was studied with an SS-OCT prototype system, which uses a tunable laser as a light source operated at 100,000 Hz A scan repetition rate in the 1-MUm wavelength region. Three-dimensional volumetric measurement comprised of 512 * 128 A scans was acquired in 0.8 second. From a series of OCT images, a chroidal thickness map of the macular area was created by manual segmentation. To evaluate interoperator reproducibility, the choroidal thickness in each section from 10 subjects was determined independently by two observers. RESULTS: SS-OCT at the 1-MUm wavelength region allowed visualization of the fine structure of the choroid as well as that of the retina. Mean choroidal thickness and volume in the macula area were, respectively, 191.5 +/- 74.2 MUm and 5.411 +/- 2.097 mm(3). The mean choroidal thickness of the outer nasal area was significantly thinner than that of all other areas (P < 0.05). The measurements by the two independent observers were significantly identical; the intraclass correlation coefficient in mean choroidal thickness was between 0.945 and 0.980 in each area. The macular choroidal thickness was significantly correlated with axial length after adjustment for age (P < 0.001), and with age after adjustment for axial length (P < 0.001). CONCLUSIONS: SS-OCT system at 1 MUm provides macular choroidal thickness maps and allows one to evaluate the choroidal thickness more accurately. PMID- 21622705 TI - Comment on "A bacterium that can grow by using arsenic instead of phosphorus". AB - Wolfe-Simon et al. (Research Articles, 3 June 2011, p. 1163; published online 2 December 2010) reported that the bacterial strain GFAJ-1 can grow by using arsenic (As) instead of phosphorus (P), noting that the P content in bacteria grown in +As/-P culture medium was far below the quantity needed to support growth. However, low P content is a common phenotype across a broad range of environmental bacteria that experience P limitation. PMID- 21622706 TI - Comment on "A bacterium that can grow by using arsenic instead of phosphorus". AB - Wolfe-Simon et al. (Research Articles, 3 June 2011, p. 1163; published online 2 December 2010) reported that bacterium GFAJ-1 can grow by using arsenic instead of phosphorus. However, the presence of contaminating phosphate in the growth medium, as well as the omission of important DNA purification steps, cast doubt on the authors' conclusion that arsenic can substitute for phosphorus in the nucleic acids of this organism. PMID- 21622707 TI - Comment on "A bacterium that can grow by using arsenic instead of phosphorus". AB - Wolfe-Simon et al. (Research Articles, 3 June 2011, p. 1163; published online 2 December 2010) argued that the bacterial strain GFAJ-1 can vary the elemental composition of its biomolecules by substituting arsenic for phosphorus. Although their data show that GFAJ-1 is an extraordinary extremophile, consideration of arsenate redox chemistry undermines the suggestion that arsenate can replace the physiologic functions of phosphate. PMID- 21622708 TI - Comment on "A bacterium that can grow by using arsenic instead of phosphorus". AB - Wolfe-Simon et al. (Research Articles, 3 June 2011, p. 1163; published online 2 December 2010) reported that bacterial strain GFAJ-1 can substitute arsenic for phosphorus in its biomolecules, including nucleic acids and proteins. Unfortunately, their study lacks crucial experimental evidence to support this claim and suffers from inadequate data and poor presentation and analysis. PMID- 21622709 TI - Comment on "A bacterium that can grow by using arsenic instead of phosphorus". AB - Wolfe-Simon et al. (Research Articles, 3 June 2011, p. 1163; published online 2 December 2010) reported that a naturally occurring bacterium, strain GFAJ-1, can substitute arsenic for phosphorus in its biomolecules. However, straightforward experiments to support this claim, including density gradient centrifugation of DNA assumed to contain arsenic, were either not performed or not presented. As a result, the authors' conclusions remain uncertain. PMID- 21622710 TI - Editor's Note. AB - The Research Article "A bacterium that can grow by using arsenic instead of phosphorus" by F. Wolfe-Simon et al., published online 2 December 2010, was the subject of extensive discussion and criticism following its online publication. Science received a wide range of correspondence that raised specific concerns about the Research Article's methods and interpretations. Eight Technical Comments that represent the main concerns, as well as a Technical Response by Wolfe-Simon et al., are published online in Science Express at the addresses listed in this note. They have been peer-reviewed and revised according to Science's standard procedure. PMID- 21622711 TI - Comment on "A bacterium that can grow by using arsenic instead of phosphorus". AB - Wolfe-Simon et al. (Research Articles, 3 June 2011, p. 1163; published online 2 December 2010) reported the discovery of an unusual bacterium, strain GFAJ-1, that can grow in the presence of high concentrations of arsenate. The authors' contention, however, that this microbe can appreciably vary the elemental composition of its fundamental biomolecules by substituting arsenic for phosphorus appears premature based on the data presented. PMID- 21622712 TI - Comment on "A bacterium that can grow by using arsenic instead of phosphorus". AB - Wolfe-Simon et al. (Research Articles, 3 June 2011, p. 1163; published online 2 December 2010) reported that bacterium GFAJ-1 can substitute arsenic for phosphorus in its macromolecules, including DNA and proteins. If such arseno-DNA exists, then much of the past century of work with arsenate and phosphate chemistry, as well as much of what we think we know about metabolism, will need rewriting. PMID- 21622713 TI - Comment on "A bacterium that can grow by using arsenic instead of phosphorus". AB - Wolfe-Simon et al. (Research Articles, 3 June 2011, p. 1163; published online 2 December 2010) reported an apparent stimulatory effect of arsenic on the growth of bacteria isolated from Mono Lake, California, which they interpreted as evidence that the cells can grow by using arsenic instead of phosphorus. Alternatively, arsenic may have stimulated the bacterium's high-affinity phosphorus assimilation pathway, which is active when phosphate levels are low. PMID- 21622714 TI - Caveolin-1 upregulation mediates suppression of primary breast tumor growth and brain metastases by stat3 inhibition. AB - Stat3 activation has been implicated as an important driver of brain metastasis in breast cancer, but the critical targets of Stat3 in this process are yet to be fully defined. In this study, we identified the lipid raft organizing protein Caveolin-1 (Cav-1) as a critical genetic target of Stat3 in this process. In human breast cancers, we found that activated Stat3 correlated with attenuation of Cav-1 in brain metastases relative to primary tumors. Cav-1 promoter activity and gene expression were increased by overexpressing an activated form of Stat3 but decreased by attenuation of Stat3 activity or expression. We identified putative Stat3-binding elements in the Cav-1 promoter and showed a direct repression of Cav-1 transcription by Stat3. Reciprocally, we showed that strategies to increase or decrease Cav-1 expression were sufficient to attenuate or promote breast cancer cell invasion. Furthermore, increased expression of Cav 1 phenocopied the effects of Stat3 activation in blocking primary tumor growth and abrogating formation of brain metastases. Collectively, our findings provide clinical and mechanistic evidence that Cav-1 is a critical target for suppression by Stat3 in driving invasion and metastasis of breast cancer cells. PMID- 21622715 TI - Sorafenib enhances pemetrexed cytotoxicity through an autophagy-dependent mechanism in cancer cells. AB - Pemetrexed (ALIMTA, Lilly) is a folate antimetabolite that has been approved by the U.S. Food and Drug Administration for the treatment of non-small cell lung cancer and has been shown to stimulate autophagy. In the present study, we sought to further understand the role of autophagy in response to pemetrexed and to test if combination therapy could enhance the level of toxicity through altered autophagy in tumor cells. The multikinase inhibitor sorafenib (Nexavar, Bayer), used in the treatment of renal and hepatocellular carcinoma, suppresses tumor angiogenesis and promotes autophagy in tumor cells. We found that sorafenib interacted in a greater than additive fashion with pemetrexed to increase autophagy and to kill a diverse array of tumor cell types. Tumor cell types that displayed high levels of cell killing after combination treatment showed elevated levels of AKT, p70 S6K, and/or phosphorylated mTOR, in addition to class III receptor tyrosine kinases such as platelet-derived growth factor receptor beta and VEGF receptors, known in vivo targets of sorafenib. In xenograft and in syngeneic animal models of mammary carcinoma and glioblastoma, the combination of sorafenib and pemetrexed suppressed tumor growth without deleterious effects on normal tissues or animal body mass. Taken together, the data suggest that premexetred and sorafenib act synergistically to enhance tumor killing via the promotion of a toxic form of autophagy that leads to activation of the intrinsic apoptosis pathway, and predict that combination treatment represents a future therapeutic option in the treatment of solid tumors. PMID- 21622716 TI - Effect of socioeconomic status on secondary prevention of stroke. AB - OBJECTIVE: Cardiovascular risk factors increase risk for stroke recurrence. Secondary prevention of stroke may be affected not only by established risk factors, but also socioeconomic status. This study evaluates relationships between socioeconomic status and cardiovascular and behavioral factors. DESIGN: Cross-section study. SETTING: Public Health and Education Institute, Peking University. PARTICIPANTS: Outpatients (n = 2354) with a past diagnosis of stroke or transient ischemic attack. INTERVENTION(S): The investigation consisted of a questionnaire regarding patients' socioeconomic and living status, and a clinical examination at the research center. MAIN OUTCOME MEASURE(S): Control rates of risk factors for cardiovascular disease. RESULTS: With regard to hypertension patients, 67.0% were aware of having hypertension, 63.6% were treated and 53.9% had controlled hypertension; for patients with hypercholesterolemia, 46.7% were aware of having hypercholesterolemia, 38.6% were treated and 3.8% had controlled hypercholesterolemia; for patients with diabetes mellitus, 28.0% were aware of having diabetes mellitus, 25.7% were treated and 3.5% had controlled diabetes mellitus. After multivariate analysis, education was the strongest associated factor for controls of hypertension and diabetes mellitus. After adjustment for sex and age, strong and graduated relationships were noted between the level of education and control of risk factors, with the odds ratios increasing at every increment. CONCLUSION: Education exerts the most important effect on the control of established cardiovascular risk factors; Successful intervention to reduce these risk factors will have to be addressed, not just with regard to specific risk factors, but also with the societal conditions that lead to the adoption and maintenance of high-risk behaviors. PMID- 21622717 TI - NGAL expression is elevated in both colorectal adenoma-carcinoma sequence and cancer progression and enhances tumorigenesis in xenograft mouse models. AB - PURPOSE: There is growing evidence implicating that neutrophil gelatinase associated lipocalin (NGAL) plays a role in the development and progression of cancers. However, the effect of NGAL in colorectal carcinoma (CRC) has not been clearly elucidated. In this study, we investigated the role of NGAL in the tumorigenesis and progression of CRC and evaluated the clinical value of NGAL expression. EXPERIMENTAL DESIGN: We examined NGAL expression in 526 colorectal tissue samples, including 53 sets of matched specimens (histologically normal mucosa, adenomas, and carcinomas) using immunohistochemical analysis. In CRCs, correlations between NGAL expression and clinicopathologic parameters were analyzed, and survival analysis was conducted. The role of NGAL was further tested using mouse xenograft models. RESULTS: NGAL expression was elevated during the colorectal adenoma-carcinoma sequence both among the 526 cases (r(s) = 0.66, P < 0.001) and in the 53 sets of matched specimens (r(s) = 0.60, P < 0.001). In CRCs, NGAL expression was associated with cancer stage (P = 0.041) and tumor recurrence in stage II patients (P = 0.037). Survival analysis revealed that NGAL expression was an independent prognostic factor for overall survival (HR = 1.84, P = 0.004) and for disease-free survival of stage II patients (HR = 5.88, P = 0.021). In mouse models, the xenografts in cecum and spleen were heavier and more numerous in the group injected with NGAL-overexpressing CRC cells (P < 0.05). CONCLUSIONS: NGAL overexpression may promote the tumorigenesis and progression of CRC. Detecting NGAL expression in tumor tissues may be useful for evaluating prognosis of patients with CRC. PMID- 21622719 TI - Multiple Loci modulate opioid therapy response for cancer pain. AB - PURPOSE: Patients treated with opioid drugs for cancer pain experience different relief responses, raising the possibility that genetic factors play a role in opioid therapy outcome. In this study, we tested the hypothesis that genetic variations may control individual response to opioid drugs in cancer patients. EXPERIMENTAL DESIGN: We tested 1 million single-nucleotide polymorphisms (SNP) in European cancer patients, selected in a first series, for extremely poor (pain relief <=40%; n = 145) or good (pain relief >=90%; n = 293) responses to opioid therapy using a DNA-pooling approach. Candidate SNPs identified by SNP-array were genotyped in individual samples constituting DNA pools as well as in a second series of 570 patients. RESULTS: Association analysis in 1,008 cancer patients identified eight SNPs significantly associated with pain relief at a statistical threshold of P < 1.0 * 10-3, with rs12948783, upstream of the RHBDF2 gene, showing the best statistical association (P = 8.1 * 10-9). Functional annotation analysis of SNP-tagged genes suggested the involvement of genes acting on processes of the neurologic system. CONCLUSION: Our results indicate that the identified SNP panel can modulate the response of cancer patients to opioid therapy and may provide a new tool for personalized therapy of cancer pain. PMID- 21622718 TI - Dual blockade of EGFR and c-Met abrogates redundant signaling and proliferation in head and neck carcinoma cells. AB - PURPOSE: Head and neck squamous cell carcinoma (HNSCC) is usually fatal, and innovative approaches targeting growth pathways are necessary to effectively treat this disease. Both the epidermal growth factor receptor (EGFR) and the hepatocyte growth factor (HGF)/c-Met pathways are overexpressed in HNSCC and initiate similar downstream signaling pathways. c-Met may act in consort with EGFR and/or be activated as a compensatory pathway in the presence of EGFR blockade. EXPERIMENTAL DESIGN: Expression levels of EGFR and c-Met were determined by Western analysis in HNSCC cell lines and correlated with antitumor responses to inhibitors of these pathways. RESULTS: Combining the c-Met inhibitor PF2341066 with the EGFR inhibitor gefitinib abrogated HNSCC cell proliferation, invasion, and wound healing significantly more than inhibition of each pathway alone in HNSCC cell lines. When both HGF and the EGFR ligand, TGF-alpha, were present in vitro, P-AKT and P-MAPK expression were maximally inhibited by targeting both EGFR and c-Met pathways, suggesting that c-Met or EGFR can compensate when phosphorylation of the other receptor is inhibited. We also showed that TGF-alpha can induce phosphorylation of c-Met over sixfold by 8 hours in the absence of HGF, supporting a ligand-independent mechanism. Combined targeting of c-Met and EGFR resulted in an enhanced inhibition of tumor volumes accompanied by a decreased number of proliferating cells and increased apoptosis compared with single agent treatment in vivo. CONCLUSIONS: Together, these results suggest that dual blockade of c-Met and EGFR may be a promising clinical therapeutic strategy for treating HNSCC. PMID- 21622720 TI - SKLB1002, a novel potent inhibitor of VEGF receptor 2 signaling, inhibits angiogenesis and tumor growth in vivo. AB - PURPOSE: VEGF receptor 2 (VEGFR2) inhibitors, as efficient antiangiogenesis agents, have been applied in the cancer treatment. However, currently most of these anticancer drugs suffer some adverse effects. Discovery of novel VEGFR2 inhibitors as anticancer drug candidates is still needed. EXPERIMENTAL DESIGN: In this investigation, we adopted a restricted de novo design method to design VEGFR2 inhibitors. We selected the most potent compound SKLB1002 and analyzed its inhibitory effects on human umbilical vein endothelial cells (HUVEC) in vitro. Tumor xenografts in zebrafish and athymic mice were used to examine the in vivo activity of SKLB1002. RESULTS: The use of the restricted de novo design method indeed led to a new potent VEGFR2 inhibitor, SKLB1002, which could significantly inhibit HUVEC proliferation, migration, invasion, and tube formation. Western blot analysis was conducted, which indicated that SKLB1002 inhibited VEGF-induced phosphorylation of VEGFR2 kinase and the downstream protein kinases including extracellular signal-regulated kinase, focal adhesion kinase, and Src. In vivo zebrafish model experiments showed that SKLB1002 remarkably blocked the formation of intersegmental vessels in zebrafish embryos. It was further found to inhibit a new microvasculature in zebrafish embryos induced by inoculated tumor cells. Finally, compared with the solvent control, administration of 100 mg/kg/d SKLB1002 reached more than 60% inhibition against human tumor xenografts in athymic mice. The antiangiogenic effect was indicated by CD31 immunohistochemical staining and alginate-encapsulated tumor cell assay. CONCLUSIONS: Our findings suggest that SKLB1002 inhibits angiogenesis and may be a potential drug candidate in anticancer therapy. PMID- 21622721 TI - The activity of trabectedin as a single agent or in combination with everolimus for clear cell carcinoma of the ovary. AB - PURPOSE: The objective of this study was to evaluate the antitumor efficacy of trabectedin in clear cell carcinoma (CCC) of the ovary, which is regarded as an aggressive, chemoresistant, histologic subtype. EXPERIMENTAL DESIGN: Using 6 human ovarian cancer cell lines (3 CCC and 3 serous adenocarcinomas), the antitumor effects of trabectedin were examined in vitro, and we compared its activity according to histology. We next examined the antitumor activity of trabectedin in both cisplatin-resistant and paclitaxel-resistant CCC cells in vitro. Then, the in vivo effects of trabectedin were evaluated using mice inoculated with CCC cell lines. Using 2 pairs of trabectedin-sensitive parental and trabectedin-resistant CCC sublines, we investigated the role of mTOR in the mechanism of acquired resistance to trabectedin. Finally, we determined the effect of mTOR inhibition by everolimus on the antitumor efficacy of trabectedin in vitro and in vivo. RESULTS: Trabectedin showed significant antitumor activity toward chemosensitive and chemoresistant CCC cells in vitro. Mouse xenografts of CCC cells revealed that trabectedin significantly inhibits tumor growth. Greater activation of mTOR was observed in trabectedin-resistant CCC cells than in their respective parental cells. The continuous inhibition of mTOR significantly enhanced the therapeutic efficacy of trabectedin and prevented CCC cells from acquiring resistance to trabectedin. CONCLUSION: Trabectedin is a promising agent for CCC as a first-line chemotherapy and as a second-line treatment of recurrent CCC that had previously been treated with cisplatin or paclitaxel. Moreover, trabectedin combined with everolimus may be more efficacious for the management of CCC. PMID- 21622722 TI - MicroRNA profiling of sporadic and hereditary medullary thyroid cancer identifies predictors of nodal metastasis, prognosis, and potential therapeutic targets. AB - PURPOSE: While the molecular basis of hereditary medullary thyroid cancer (HMTC) has been well defined, little is known about the molecular pathogenesis of sporadic medullary thyroid cancer (SMTC). In addition, microRNAs (miRNAs) have been shown to be important diagnostic and prognostic markers in cancer but have not been defined in MTC. Our aim was to study the miRNA profile of MTC to identify prognostic biomarkers and potential therapeutic targets. EXPERIMENTAL DESIGN: MiRNA microarray profiling was carried out in fresh frozen tissues from patients with SMTC (n = 12) and HMTC (n = 7). Differential expression of three miRNAs was confirmed in a validation cohort of SMTC and HMTC samples (n = 45) using quantitative reverse transcriptase-PCR and correlated with clinical outcomes. The functional role of a selected miRNA was investigated in vitro in the human medullary thyroid carcinoma cell line (TT cells) using cell proliferation assays and Western blotting analysis. RESULTS: MiRs-183 and 375 were overexpressed (P = 0.001; 0.031) and miR-9* was under-expressed (P = 0.011) in SMTC versus HMTC. Overexpression of miRs-183 and 375 in MTC predicted lateral lymph node metastases (P < 0.001; P = 0.001) and was associated with residual disease (P = 0.001; 0.003), distant metastases (P = 0.003; 0.001), and mortality (P = 0.01; 0.011). Knock down of miR-183 expression in the TT cell line induced a significant decrease in the viable cell count and upregulation of the protein LC3B, which is associated with autophagy. CONCLUSIONS: Our data indicate that miRNAs play a pivotal role in the biology of MTC and represent an important class of prognostic biomarkers and therapeutic targets warranting further investigation. PMID- 21622723 TI - Cancer stem cells and chemosensitivity. AB - Cancer lethality is mainly due to the onset of distant metastases and refractoriness to chemotherapy. Thus, the development of molecular targeted agents that can restore or increase chemosensitivity will provide valuable therapeutic options for cancer patients. Growing evidence indicates that a cellular subpopulation with stem cell-like features, commonly referred to as cancer stem cells (CSCs), is critical for tumor generation and maintenance. Recent advances in stem cell biology are revealing that this cellular fraction shares many properties with normal adult stem cells and is able to propagate the parental tumor in animal models. CSCs seem to be protected against widely used chemotherapeutic agents by means of different mechanisms, such as a marked proficiency in DNA damage repair, high expression of ATP-binding cassette drug transporters, and activation of PI3K/AKT and Wnt pathways. Moreover, microenvironmental stimuli such as those involved in the epithelial-mesenchymal transition and hypoxia indirectly contribute to chemoresistance by inducing in cancer cells a stem-like phenotype. Understanding how CSCs overcome chemotherapy induced death stimuli, and integrating such knowledge into clinical research methodology, has become a priority in the process of identifying innovative therapeutic strategies aimed at improving the outcome of cancer patients. PMID- 21622724 TI - FGFR-4 Arg388 enhances prostate cancer progression via extracellular signal related kinase and serum response factor signaling. AB - PURPOSE: Increased expression of FGFR-4 and its ligands have been linked to lethal prostate cancer (PCa). Furthermore, a germ line polymorphism in the FGFR-4 gene, resulting in arginine at codon 388 (Arg388) instead of glycine (Gly388), is associated with aggressive disease. The FGFR-4 Arg388 variant results in increased receptor stability, sustained receptor activation, and increased motility and invasion compared with Gly388. However, the impact of sustained signaling on cellular signal transduction pathways is unknown. EXPERIMENTAL DESIGN: Expression microarray analysis of immortalized prostatic epithelial cells lines expressing FGFR-4 Arg388 or Gly388 was used to establish a gene signature associated with FGFR-4 Arg388 expression. Transient transfection of reporters and inhibitors was used to establish the pathways activated by FGFR-4 Arg388 expression. The impact of pathway knockdown in vitro and in an orthotopic model was assessed using inhibitors and/or short hairpin RNA (shRNA). RESULTS: Expression of the FGFR-4 Arg388 protein leads to increased activity of the extracellular signal-related kinase (ERK) pathway, increased activity of serum response factor (SRF) and AP1, and transcription of multiple genes that are correlated with aggressive clinical behavior in PCa. Increased expression of SRF is associated with biochemical recurrence in men undergoing radical prostatectomy. Consistent with these observations, knockdown of FGFR-4 Arg388 in PCa cells decreases proliferation and invasion in vitro and primary tumor growth and metastasis in vivo. CONCLUSIONS: These studies define a signal transduction pathway downstream of FGFR-4 Arg388 that acts via ERK and SRF to promote PCa progression. PMID- 21622725 TI - Brivanib, a dual FGF/VEGF inhibitor, is active both first and second line against mouse pancreatic neuroendocrine tumors developing adaptive/evasive resistance to VEGF inhibition. AB - PURPOSE: Preclinical trials of a mouse model of pancreatic neuroendocrine tumors (PNET) were conducted to determine whether dual FGF/VEGF pathway inhibition with brivanib can improve first-line efficacy in comparison with VEGF inhibitors lacking fibroblast growth factor (FGF)-inhibitory activity and to characterize second-line brivanib activity before and after the onset of evasive resistance to VEGF-selective therapy. EXPERIMENTAL DESIGN: An anti-VEGFR2 monoclonal antibody (DC101), an inhibitor of FGF signaling (FGF ligand trap), sorafenib, and brivanib were comparatively evaluated in first-line monotherapy in short and longer term fixed endpoint intervention trials in the RIP-Tag2 mouse model of PNET. Brivanib was also tested second line aiming to block adaptive resistance to selective VEGF therapies, assessing tumor growth, vascularity, hypoxia, invasion, and metastasis. The effects of initiating second-line brivanib therapy prior to or following overt relapse on sorafenib therapy were compared in overall survival trials to first-line therapies. RESULTS: Brivanib produced enduring tumor stasis and angiogenic blockade, both first and second line following the failure of DC101 or sorafenib. Overall survival was significantly extended by brivanib versus sorafenib, both first-line and when second-line therapy was initiated prior to sorafenib failure; second-line brivanib was less beneficial when initiated later, after the initiation of revascularization and incipient tumor progression. CONCLUSIONS: Brivanib holds promise and deserves consideration for clinical evaluation as an antiangiogenic therapy, both in the context of impending failures of VEGF-selective therapy and in a first-line setting aiming to limit the adaptive response to VEGF inhibitors that results in evasive resistance. PMID- 21622726 TI - Metabolism of free oligosaccharides is facilitated in the och1Delta mutant of Saccharomyces cerevisiae. AB - In eukaryotic cells, it is known that N-glycans play a pivotal role in quality control of carrier proteins. Although "free" forms of oligosaccharides (fOSs) are known to be accumulated in the cytosol, the precise mechanism of their formation, degradation and biological relevance remains poorly understood. It has been shown that, in budding yeast, almost all fOSs are formed from misfolded glycoproteins. Precise structural analysis of fOSs revealed that several yeast fOSs bear a yeast specific modification by Golgi-resident alpha-1,6-mannosyltransferase, Och1. In this study, structural diversity of fOSs in och1Delta cells was analyzed. To our surprise, several fOSs in och1Delta cells have unusual alpha-1,3-linked mannose residues at their non-reducing termini. These mannose residues were not observed in wild-type cells, suggesting that the addition of these unique mannoses occurred as a compensation of Och1 defect. A significant increase in the amount of fOSs modified by Golgi-localized mannosyltransferases was also observed in och1Delta cells. Moreover, the amount of processed fOSs and intracellular alpha mannosidase (Ams1) both increased in this mutant. Up-regulation of Ams1 activity was also apparent for cells treated with cell wall perturbation reagent. These results provide an insight into a possible link between catabolism of fOSs and cell wall stress. PMID- 21622727 TI - Inflammation and increased aromatase expression occur in the breast tissue of obese women with breast cancer. AB - Obesity is a risk factor for the development of hormone receptor-positive breast cancer in postmenopausal women and has been associated with an increased risk of recurrence and reduced survival. In humans, obesity causes subclinical inflammation in visceral and subcutaneous adipose tissue, characterized by necrotic adipocytes surrounded by macrophages forming crown-like structures (CLS). Recently, we found increased numbers of CLS, activation of the NF-kappaB transcription factor, and elevated aromatase levels and activity in the mammary glands of obese mice. These preclinical findings raised the possibility that the obesity -> inflammation axis is important for the development and progression of breast cancer. Here, our main objective was to determine if the findings in mouse models of obesity translated to women. Breast tissue was obtained from 30 women who underwent breast surgery. CLS of the breast (CLS-B) was found in nearly 50% (14 of 30) of patient samples. The severity of breast inflammation, defined as the CLS-B index, correlated with both body mass index (P < 0.001) and adipocyte size (P = 0.01). Increased NF-kappaB binding activity and elevated aromatase expression and activity were found in the inflamed breast tissue of overweight and obese women. Collectively, our results suggest that the obesity -> inflammation -> aromatase axis is present in the breast tissue of most overweight and obese women. The presence of CLS-B may be a biomarker of increased breast cancer risk or poor prognosis. PMID- 21622728 TI - Characterization of a genomic signature of pregnancy identified in the breast. AB - The objective of this study was to comprehensively compare the genomic profiles in the breast of parous and nulliparous postmenopausal women to identify genes that permanently change their expression following pregnancy. The study was designed as a two-phase approach. In the discovery phase, we compared breast genomic profiles of 37 parous with 18 nulliparous postmenopausal women. In the validation phase, confirmation of the genomic patterns observed in the discovery phase was sought in an independent set of 30 parous and 22 nulliparous postmenopausal women. RNA was hybridized to Affymetrix HG_U133 Plus 2.0 oligonucleotide arrays containing probes to 54,675 transcripts, scanned and the images analyzed using Affymetrix GCOS software. Surrogate variable analysis, logistic regression, and significance analysis of microarrays were used to identify statistically significant differences in expression of genes. The false discovery rate (FDR) approach was used to control for multiple comparisons. We found that 208 genes (305 probe sets) were differentially expressed between parous and nulliparous women in both discovery and validation phases of the study at an FDR of 10% and with at least a 1.25-fold change. These genes are involved in regulation of transcription, centrosome organization, RNA splicing, cell-cycle control, adhesion, and differentiation. The results provide initial evidence that full-term pregnancy induces long-term genomic changes in the breast. The genomic signature of pregnancy could be used as an intermediate marker to assess potential chemopreventive interventions with hormones mimicking the effects of pregnancy for prevention of breast cancer. PMID- 21622729 TI - Eye guidance in natural vision: reinterpreting salience. AB - Models of gaze allocation in complex scenes are derived mainly from studies of static picture viewing. The dominant framework to emerge has been image salience, where properties of the stimulus play a crucial role in guiding the eyes. However, salience-based schemes are poor at accounting for many aspects of picture viewing and can fail dramatically in the context of natural task performance. These failures have led to the development of new models of gaze allocation in scene viewing that address a number of these issues. However, models based on the picture-viewing paradigm are unlikely to generalize to a broader range of experimental contexts, because the stimulus context is limited, and the dynamic, task-driven nature of vision is not represented. We argue that there is a need to move away from this class of model and find the principles that govern gaze allocation in a broader range of settings. We outline the major limitations of salience-based selection schemes and highlight what we have learned from studies of gaze allocation in natural vision. Clear principles of selection are found across many instances of natural vision and these are not the principles that might be expected from picture-viewing studies. We discuss the emerging theoretical framework for gaze allocation on the basis of reward maximization and uncertainty reduction. PMID- 21622730 TI - Restitution of tumor suppressor microRNAs using a systemic nanovector inhibits pancreatic cancer growth in mice. AB - Mis-expression of microRNAs (miRNA) is widespread in human cancers, including in pancreatic cancer. Aberrations of miRNA include overexpression of oncogenic miRs (Onco-miRs) or downregulation of so-called tumor suppressor TSG-miRs. Restitution of TSG-miRs in cancer cells through systemic delivery is a promising avenue for pancreatic cancer therapy. We have synthesized a lipid-based nanoparticle for systemic delivery of miRNA expression vectors to cancer cells (nanovector). The plasmid DNA-complexed nanovector is approximately 100 nm in diameter and shows no apparent histopathologic or biochemical evidence of toxicity upon intravenous injection. Two miRNA candidates known to be downregulated in the majority of pancreatic cancers were selected for nanovector delivery: miR-34a, which is a component of the p53 transcriptional network and regulates cancer stem cell survival, and the miR-143/145 cluster, which together repress the expression of KRAS2 and its downstream effector Ras-responsive element binding protein-1 (RREB1). Systemic intravenous delivery with either miR-34a or miR-143/145 nanovectors inhibited the growth of MiaPaCa-2 subcutaneous xenografts (P < 0.01 for miR-34a; P < 0.05 for miR-143/145); the effects were even more pronounced in the orthotopic (intrapancreatic) setting (P < 0.0005 for either nanovector) when compared with vehicle or mock nanovector delivering an empty plasmid. Tumor growth inhibition was accompanied by increased apoptosis and decreased proliferation. The miRNA restitution was confirmed in treated xenografts by significant upregulation of the corresponding miRNA and significant decreases in specific miRNA targets (SIRT1, CD44 and aldehyde dehydrogenase for miR34a, and KRAS2 and RREB1 for miR-143/145). The nanovector is a platform with potential broad applicability in systemic miRNA delivery to cancer cells. PMID- 21622731 TI - Trabectedin and its C subunit modified analogue PM01183 attenuate nucleotide excision repair and show activity toward platinum-resistant cells. AB - PM01183 is a novel marine-derived covalent DNA binder in clinical development. PM01183 is structurally similar to trabectedin (yondelis, ecteinascidin-743) except for the C subunit, and this modification is accompanied by different pharmacokinetics in cancer patients. We here characterize the interaction of PM01183 with the nucleotide excision repair (NER) pathway in comparison with trabectedin. Our results show for the first time that although neither PM01183 nor trabectedin is repaired by NER, both compounds are able to interfere with the NER machinery thereby attenuating the repair of specific NER substrates. We further show that the NER activity is increased in 3 of 4 cellular models with acquired resistance to cisplatin or oxaliplatin, confirming the involvement of NER in the resistance to platinum derivatives. Importantly, both PM01183 and trabectedin show unchanged or even enhanced activity toward all 4 cisplatin- and oxaliplatin-resistant cell lines. We finally show that combinations of PM01183 and cisplatin were mostly synergistic toward both parental and cisplatin resistant ovarian carcinoma cells as indicated by Chou and Talalay analysis. These data show that the C subunit of trabectedin can be subjected to at least some structural modifications without loss of activity or NER interaction. While PM01183 and trabectedin appear functionally similar in cellular models, it is likely that the differences in pharmacokinetics may allow different dosing and scheduling of PM01183 in the clinic that could lead to novel and/or increased antitumor activity. Taken together, our results provide a mechanistic basis to support clinical trials of PM01183 alone or in combination with cisplatin. PMID- 21622732 TI - Delayed administration of suramin attenuates the progression of renal fibrosis in obstructive nephropathy. AB - We recently showed that suramin treatment prevents the onset of renal fibrosis in a model of obstructive nephropathy induced by unilateral ureteral obstruction (UUO). In this study, we further assessed the effect of delayed administration of suramin on the progression of tubulointerstitial fibrosis. Mice were given a single dose of suramin at 20 mg/kg starting at day 3 of obstruction, and kidneys were harvested after an additional 7 or 14 days of obstruction. Suramin completely blocked further increase in expression of type I collagen and fibronectin and largely suppressed expression of alpha-smooth muscle actin (alpha SMA) in both treatment groups. UUO injury induced phosphorylation of Smad-3, a key mediator of transforming growth factor-beta (TGF-beta) signaling, epidermal growth factor receptor, and platelet-derived growth factor receptor after 3 days and further increased at 10 days after UUO injury. When suramin was administered at 3 days after obstruction, phosphorylation of these molecules was not further increased in the obstructed kidney. Suramin treatment also inhibited activation of signal transducer and activator of transcription 3 and extracellular signal regulated kinase 1 and 2, two signaling pathways associated with renal fibrogenesis. Furthermore, delayed application of suramin suppressed TGF-beta1 induced expression of alpha-SMA and fibronectin in cultured renal interstitial fibroblasts. These results indicate that administration of suramin is effective in attenuating the progression of renal fibrosis after injury and suggest the potential clinical application of suramin as an antifibrotic treatment in patients with chronic kidney disease. PMID- 21622733 TI - Inflammatory stimuli inhibit glucocorticoid-dependent transactivation in human pulmonary epithelial cells: rescue by long-acting beta2-adrenoceptor agonists. AB - By repressing inflammatory gene expression, glucocorticoids are the most effective treatment for chronic inflammatory diseases such as asthma. However, in some patients with severe disease, or who smoke or suffer from chronic obstructive pulmonary disease, glucocorticoids are poorly effective. Although many investigators focus on defects in the repression of inflammatory gene expression, glucocorticoids also induce (transactivate) the expression of numerous genes to elicit anti-inflammatory effects. Using human bronchial epithelial (BEAS-2B) and pulmonary (A549) cells, we show that cytokines [tumor necrosis factor alpha (TNFalpha) and interleukin 1beta], mitogens [fetal calf serum (FCS) and phorbol ester], cigarette smoke, and a G(q)-linked G protein coupled receptor agonist attenuate simple glucocorticoid response element (GRE) dependent transcription. With TNFalpha and FCS, this effect was not overcome by increasing concentrations of dexamethasone, budesonide, or fluticasone propionate. Thus, the maximal ability of the glucocorticoid to promote GRE dependent transcription was reduced, and this was shown additionally for the glucocorticoid-induced gene p57(KIP2). The long-acting beta(2)-adrenoceptor agonists (LABAs) formoterol fumarate and salmeterol xinafoate enhanced simple GRE dependent transcription to a level that could not be achieved by glucocorticoid alone. In the presence of TNFalpha or FCS, which repressed glucocorticoid responsiveness, these LABAs restored glucocorticoid-dependent transcription to levels that were achieved by glucocorticoid alone. Given the existence of genes, such as p57(KIP2), which may mediate anti-inflammatory actions of glucocorticoids, we propose that repression of transactivation represents a mechanism for glucocorticoid resistance and for understanding the clinical benefit of LABAs as an add-on therapy in asthma and chronic obstructive pulmonary disease. PMID- 21622734 TI - Quorum sensing in Chromobacterium violaceum: DNA recognition and gene regulation by the CviR receptor. AB - The bacterial pathogen Chromobacterium violaceum uses a LuxIR-type quorum-sensing system to detect and respond to changes in cell population density. CviI synthesizes the autoinducer C(10)-homoserine lactone (C(10)-HSL), and CviR is a cytoplasmic DNA binding transcription factor that activates gene expression following binding to C(10)-HSL. A number of behaviors are controlled by quorum sensing in C. violaceum. However, few genes have been shown to be directly controlled by CviR, in part because the DNA motif bound by CviR is not well characterized. Here, we define the DNA sequence required for promoter recognition by CviR. Using in vivo data generated from a library of point mutations in a CviR regulated promoter, we find that CviR binds to a palindrome with the ideal sequence CTGNCCNNNNGGNCAG. We constructed a position weight matrix using these in vivo data and scanned the C. violaceum genome to predict CviR binding sites. We measured direct activation of the identified promoters by CviR and found that CviR controls the expression of the promoter for a chitinase, a type VI secretion related gene, a transcriptional regulator gene, a guanine deaminase gene, and cviI. Indeed, regulation of cviI expression by CviR generates a canonical quorum sensing positive-feedback loop. PMID- 21622735 TI - Genome sequence of Rhodobacter sphaeroides Strain WS8N. AB - Rhodobacter sphaeroides is a metabolically diverse photosynthetic alphaproteobacterium found ubiquitously in soil and freshwater habitats. Here we present the annotated genome sequence of R. sphaeroides WS8N. PMID- 21622736 TI - Phosphorylation of Spo0A by the histidine kinase KinD requires the lipoprotein med in Bacillus subtilis. AB - The response regulatory protein Spo0A of Bacillus subtilis is activated by phosphorylation by multiple histidine kinases via a multicomponent phosphorelay. Here we present evidence that the activity of one of the kinases, KinD, depends on the lipoprotein Med, a mutant of which has been known to cause a cannibalism phenotype. We show that the absence of Med impaired and the overproduction of Med stimulated the transcription of two operons (sdp and skf) involved in cannibalism whose transcription is known to depend on Spo0A in its phosphorylated state (Spo0A~P). Further, these effects of Med were dependent on KinD but not on kinases KinA, KinB, and KinC. Additionally, we show that deletion or overproduction of Med impaired or enhanced, respectively, biofilm formation and that these effects, too, depended specifically on KinD. Finally, we report that overproduction of Med bypassed the dominant negative effect on transcription of sdp of a truncated KinD retaining the transmembrane segments but lacking the kinase domain. We propose that Med directly or indirectly interacts with KinD in the cytoplasmic membrane and that this interaction is required for KinD-dependent phosphorylation of Spo0A. PMID- 21622737 TI - Role of high-fidelity Escherichia coli DNA polymerase I in replication bypass of a deoxyadenosine DNA-peptide cross-link. AB - Reaction of bifunctional electrophiles with DNA in the presence of peptides can result in DNA-peptide cross-links. In particular, the linkage can be formed in the major groove of DNA via the exocyclic amino group of adenine (N6-dA). We previously demonstrated that an A family human polymerase, Pol nu, can efficiently and accurately synthesize DNA past N6-dA-linked peptides. Based on these results, we hypothesized that another member of that family, Escherichia coli polymerase I (Pol I), may also be able to bypass these large major groove DNA lesions. To test this, oligodeoxynucleotides containing a site-specific N6-dA dodecylpeptide cross-link were created and utilized for in vitro DNA replication assays using E. coli DNA polymerases. The results showed that Pol I and Pol II could efficiently and accurately bypass this adduct, while Pol III replicase, Pol IV, and Pol V were strongly inhibited. In addition, cellular studies were conducted using E. coli strains that were either wild type or deficient in all three DNA damage-inducible polymerases, i.e., Pol II, Pol IV, and Pol V. When single-stranded DNA vectors containing a site-specific N6-dA dodecylpeptide cross link were replicated in these strains, the efficiencies of replication were comparable, and in both strains, intracellular bypass of the lesion occurred in an error-free manner. Collectively, these findings demonstrate that despite its constrained active site, Pol I can catalyze DNA synthesis past N6-dA-linked peptide cross-links and is likely to play an essential role in cellular bypass of large major groove DNA lesions. PMID- 21622738 TI - Genome sequence of the vertebrate gut symbiont Lactobacillus reuteri ATCC 53608. AB - Lactobacillus reuteri, inhabiting the gastrointestinal tracts of a range of vertebrates, is a true symbiont with effects established as beneficial to the host. Here we describe the draft genome of L. reuteri ATCC 53608, isolated from a pig. The genome sequence provides important insights into the evolutionary changes underlying host specialization. PMID- 21622739 TI - Genome sequence of Lactococcus garvieae 21881, isolated in a case of human septicemia. AB - Lactococcus garvieae is a Gram-positive bacterium considered an important opportunistic emerging human pathogen and also a well-recognized fish pathogen. Here, we present the draft genome sequence of Lactococcus garvieae strain 21881 (2,164,557 bp, with a G+C content of 37.9%), which represents the first report of a genome sequence on Lactococcus garvieae. PMID- 21622740 TI - Genetic variation of adenylation domains of the anabaenopeptin synthesis operon and evolution of substrate promiscuity. AB - Anabaenopeptins (AP) are bioactive cyclic hexapeptides synthesized nonribosomally in cyanobacteria. APs are characterized by several conserved motifs, including the ureido bond, N-methylation in position 5, and d-Lys in position 2. All other positions of the AP molecule are variable, resulting in numerous structural variants. We have identified a nonribosomal peptide synthetase (NRPS) operon from Planktothrix agardhii strain CYA126/8 consisting of five genes (apnA to apnE) encoding six NRPS modules and have confirmed its role in AP synthesis by the generation of a mutant via insertional inactivation of apnC. In order to correlate the genetic diversity among adenylation domains (A domains) with AP structure variation, we sequenced the A domains of all six NRPS modules from seven Planktothrix strains differing in the production of AP congeners. It is remarkable that single strains coproduce APs bearing either of the chemically divergent amino acids Arg and Tyr in exocyclic position 1. Since the A domain of the initiation module (the ApnA A1 domain) has been proposed to activate the amino acid incorporated into exocyclic position 1, we decided to analyze this domain both biochemically and phylogenetically. Only ApnA A1 enzymes from strains producing AP molecules containing Arg or Tyr in position 1 were found to activate these two chemically divergent amino acids in vitro. Phylogenetic analysis of apn A domain sequences revealed that strains with a promiscuous ApnA A1 domain are derived from an ancestor that activates only Arg. Surprisingly, positive selection appears to affect only three codons within the apnA A1 gene, suggesting that this remarkable promiscuity has evolved from point mutations only. PMID- 21622741 TI - Genome sequences of the zoonotic pathogens Chlamydia psittaci 6BC and Cal10. AB - Chlamydia psittaci is a highly prevalent avian pathogen and the cause of a potentially lethal zoonosis, causing life-threatening pneumonia in humans. We report the genome sequences of C. psittaci 6BC, the prototype strain of the species, and C. psittaci Cal10, a widely used laboratory strain. PMID- 21622742 TI - TdaA regulates Tropodithietic acid synthesis by binding to the tdaC promoter region. AB - Silicibacter sp. TM1040, a member of the marine Roseobacter clade, produces the antibiotic and quorum signaling molecule tropodithietic acid (TDA), encoded by tdaABCDEF. Here, we showed that an LysR-type transcriptional regulator, TdaA, is a positive regulator of tdaCDE gene expression and binds to the tdaC promoter region. PMID- 21622743 TI - Using single-nucleotide polymorphisms to discriminate disease-associated from carried genomes of Neisseria meningitidis. AB - Neisseria meningitidis is one of the main agents of bacterial meningitis, causing substantial morbidity and mortality worldwide. However, most of the time N. meningitidis is carried as a commensal not associated with invasive disease. The genomic basis of the difference between disease-associated and carried isolates of N. meningitidis may provide critical insight into mechanisms of virulence, yet it has remained elusive. Here, we have taken a comparative genomics approach to interrogate the difference between disease-associated and carried isolates of N. meningitidis at the level of individual nucleotide variations (i.e., single nucleotide polymorphisms [SNPs]). We aligned complete genome sequences of 8 disease-associated and 4 carried isolates of N. meningitidis to search for SNPs that show mutually exclusive patterns of variation between the two groups. We found 63 SNPs that distinguish the 8 disease-associated genomes from the 4 carried genomes of N. meningitidis, which is far more than can be expected by chance alone given the level of nucleotide variation among the genomes. The putative list of SNPs that discriminate between disease-associated and carriage genomes may be expected to change with increased sampling or changes in the identities of the isolates being compared. Nevertheless, we show that these discriminating SNPs are more likely to reflect phenotypic differences than shared evolutionary history. Discriminating SNPs were mapped to genes, and the functions of the genes were evaluated for possible connections to virulence mechanisms. A number of overrepresented functional categories related to virulence were uncovered among SNP-associated genes, including genes related to the category "symbiosis, encompassing mutualism through parasitism." PMID- 21622744 TI - Complete genome sequence of Bordetella pertussis CS, a Chinese pertussis vaccine strain. AB - Bordetella pertussis is the causative agent of pertussis. Here, we report the genome sequence of Bordetella pertussis strain CS, isolated from an infant patient in Beijing and widely used as a vaccine strain for production of an acellular pertussis vaccine in China. PMID- 21622745 TI - Genomes of three methylotrophs from a single niche reveal the genetic and metabolic divergence of the methylophilaceae. AB - The genomes of three representatives of the family Methylophilaceae, Methylotenera mobilis JLW8, Methylotenera versatilis 301, and Methylovorus glucosetrophus SIP3-4, all isolated from a single study site, Lake Washington in Seattle, WA, were completely sequenced. These were compared to each other and to the previously published genomes of Methylobacillus flagellatus KT and an unclassified Methylophilales strain, HTCC2181. Comparative analysis revealed that the core genome of Methylophilaceae may be as small as approximately 600 genes, while the pangenome may be as large as approximately 6,000 genes. Significant divergence between the genomes in terms of both gene content and gene and protein conservation was uncovered, including the varied presence of certain genes involved in methylotrophy. Overall, our data demonstrate that metabolic potentials can vary significantly between different species of Methylophilaceae, including organisms inhabiting the very same environment. These data suggest that genetic divergence among the members of this family may be responsible for their specialized and nonredundant functions in C1 cycling, which in turn suggests means for their successful coexistence in their specific ecological niches. PMID- 21622746 TI - Genome sequence of Pseudomonas sp. Strain S9, an extracellular arylsulfatase producing bacterium isolated from Mangrove Soil. AB - Pseudomonas sp. strain S9 was originally isolated from mangrove soil in Xiamen, China. It is an aerobic bacterium which shows extracellular arylsulfatase activity. Here, we describe the 4.8-Mb draft genome sequence of Pseudomonas sp. S9, which exhibits novel cysteine-type sulfatases. PMID- 21622747 TI - Complete genome sequence of the universal killer Salmonella enterica Serovar Typhimurium UK-1 (ATCC 68169). AB - The Salmonella enterica serovar Typhimurium strain UK-1 exhibits the highest invasion and virulence attributes among the most frequently studied strains. S. Typhimurium UK-1 has been used as the foundation for developing recombinant vaccines and has been used extensively on virulence and colonization studies in chickens and mice. We describe here the complete genome sequence of S. Typhimurium UK-1. Comparative genomics of Salmonella Typhimurium will provide insight into factors that determine virulence and invasion. PMID- 21622748 TI - The riboswitch regulates a thiamine pyrophosphate ABC transporter of the oral spirochete Treponema denticola. AB - Thiamine pyrophosphate (TPP), a biologically active form of thiamine (vitamin B1), is an essential cofactor in all living systems. Microorganisms either synthesize TPP via de novo biosynthesis pathways or uptake exogenous thiamine from the environment via specific transporters. The oral spirochete Treponema denticola is an important pathogen that is associated with human periodontal diseases. It lacks a de novo TPP biosynthesis pathway and needs exogenous TPP for growth, suggesting that it may obtain exogenous TPP via a thiamine transporter. In this study, we identified a gene cluster that encodes a TPP ABC transporter which consists of a TPP-binding protein (TDE0143), a transmembrane permease (TDE0144), and a cytosolic ATPase (TDE0145). Transcriptional and translational analyses showed that the genes encoding these three proteins are cotranscribed and form an operon (tbpABC(Td)) that is initiated by a sigma70-like promoter. The expression level of this operon is negatively regulated by exogenous TPP and is mediated by a TPP-sensing riboswitch (Td(thi-)(box)). Genetic and biochemical studies revealed that the TDE0143 deletion mutant (T. denticola DeltatbpA) had a decreased ability to transport exogenous TPP, and the mutant failed to grow when exogenous TPP was insufficient. These results taken together indicate that the tbpABC(Td) operon encodes an ABC transporter that is required for the uptake of exogenous TPP and that the expression of this operon is regulated by a TPP binding riboswitch via a feedback inhibition mechanism. PMID- 21622749 TI - Lateral transfer of genes and gene fragments in Staphylococcus extends beyond mobile elements. AB - The widespread presence of antibiotic resistance and virulence among Staphylococcus isolates has been attributed in part to lateral genetic transfer (LGT), but little is known about the broader extent of LGT within this genus. Here we report the first systematic study of the modularity of genetic transfer among 13 Staphylococcus genomes covering four distinct named species. Using a topology-based phylogenetic approach, we found, among 1,354 sets of homologous genes examined, strong evidence of LGT in 368 (27.1%) gene sets, and weaker evidence in another 259 (19.1%). Within-gene and whole-gene transfer contribute almost equally to the topological discordance of these gene sets against a reference phylogeny. Comparing genetic transfer in single-copy and in multicopy gene sets, we observed a higher frequency of LGT in the latter, and a substantial functional bias in cases of whole-gene transfer (little such bias was observed in cases of fragmentary genetic transfer). We found evidence that lateral transfer, particularly of entire genes, impacts not only functions related to antibiotic, drug, and heavy-metal resistance, as well as membrane transport, but also core informational and metabolic functions not associated with mobile elements. Although patterns of sequence similarity support the cohesion of recognized species, LGT within S. aureus appears frequently to disrupt clonal complexes. Our results demonstrate that LGT and gene duplication play important parts in functional innovation in staphylococcal genomes. PMID- 21622750 TI - Functional characterization of seven gamma-Glutamylpolyamine synthetase genes and the bauRABCD locus for polyamine and beta-Alanine utilization in Pseudomonas aeruginosa PAO1. AB - Pseudomonas aeruginosa and many other bacteria can utilize biogenic polyamines, including diaminopropane (DAP), putrescine (Put), cadaverine (Cad), and spermidine (Spd), as carbon and/or nitrogen sources. Transcriptome analysis in response to exogenous Put and Spd led to the identification of a list of genes encoding putative enzymes for the catabolism of polyamines. Among them, pauA1 to pauA6, pauB1 to pauB4, pauC, and pauD1 and pauD2 (polyamine utilization) encode enzymes homologous to Escherichia coli PuuABCD of the gamma-glutamylation pathway in converting Put into GABA. A series of unmarked pauA mutants was constructed for growth phenotype analysis. The results revealed that it requires specific combinations of pauA knockouts to abolish utilization of different polyamines and support the importance of gamma-glutamylation for polyamine catabolism in P. aeruginosa. Another finding was that the list of Spd-inducible genes overlaps almost completely with that of Put-inducible ones except the pauA3B2 operon and the bauABCD operon (beta-alanine utilization). Mutation analysis led to the conclusion that pauA3B2 participate in catabolism of DAP, which is related to the aminopropyl moiety of Spd, and that bauABCD are essential for growth on beta alanine derived from DAP (or Spd) catabolism via the gamma-glutamylation pathway. Measurements of the pauA3-lacZ and bauA-lacZ expression indicated that these two promoters were differentially induced by Spd, DAP, and beta-alanine but showed no apparent response to Put, Cad, and GABA. Induction of the pauA3 and bauA promoters was abolished in the bauR mutant. The recombinant BauR protein was purified to demonstrate its interactions with the pauA3 and bauA regulatory regions in vitro. In summary, the present study support that the gamma glutamylation pathway for polyamine utilization is evolutionarily conserved in E. coli and Pseudomonas spp. and is further expanded in Pseudomonas to accommodate a more diverse metabolic capacity in this group of microorganisms. PMID- 21622751 TI - Complete genome sequence of Lactobacillus buchneri NRRL B-30929, a novel strain from a commercial ethanol plant. AB - Lactobacillus buchneri strain NRRL B-30929 was a contaminant obtained from a commercial ethanol fermentation. This facultative anaerobe is unique because of its rapid growth on xylose and simultaneous fermentation of xylose and glucose. The strain utilizes a broad range of carbohydrate substrates and possesses a high tolerance to ethanol and other stresses, making it an attractive candidate for bioconversion of biomass substrates to various bioproducts. The genome sequence of NRRL B-30929 will provide insight into the unique properties of this lactic acid bacterium. PMID- 21622752 TI - Comparative genomic analysis of the hexuronate metabolism genes and their regulation in gammaproteobacteria. AB - The hexuronate metabolism in Escherichia coli is regulated by two related transcription factors from the FadR subfamily of the GntR family, UxuR and ExuR. UxuR controls the d-glucuronate metabolism, while ExuR represses genes involved in the metabolism of all hexuronates. We use a comparative genomics approach to reconstruct the hexuronate metabolic pathways and transcriptional regulons in gammaproteobacteria. We demonstrate differences in the binding motifs of UxuR and ExuR, identify new candidate members of the UxuR/ExuR regulons, and describe the links between the UxuR/ExuR regulons and the adjacent regulons UidR, KdgR, and YjjM. We provide experimental evidence that two predicted members of the UxuR regulon, yjjM and yjjN, are the subject of complex regulation by this transcription factor in E. coli. PMID- 21622753 TI - Complete genome sequence of Pusillimonas sp. T7-7, a cold-tolerant diesel oil degrading bacterium isolated from the Bohai Sea in China. AB - Pusillimonas sp. T7-7 is a diesel oil-degrading cold-tolerant bacterium isolated from the benthal mud of a petroleum-contaminated site in Bohai Sea, China. We present here the complete genome sequence of T7-7. Genome analysis revealed many features of typical marine bacteria, including the absence of intact sugar metabolic pathways, the presence of glyoxylate and gluconeogenesis pathways, and the abilities for nitrate assimilation and denitrification, as well as sulfate reduction and sulfite oxidation. The presence of novel genes for the degradation of diesel oils was suggested. PMID- 21622754 TI - Genome sequence of the algicidal bacterium Kordia algicida OT-1. AB - Kordia algicida OT-1 is an algicidal bacterium against the bloom-forming microalgae. The genome sequence of K. algicida revealed a number of interesting features, including the degradation of macromolecules, the biosynthesis of carotenoid pigment and secondary metabolites, and the capacity for gliding motility, which might facilitate the understanding of algicidal mechanisms. PMID- 21622755 TI - Complete genome sequence of Melissococcus plutonius ATCC 35311. AB - We report the first completely annotated genome sequence of Melissococcus plutonius ATCC 35311. M. plutonius is a one-genus, one-species bacterium and the etiological agent of European foulbrood of the honeybee. The genome sequence will provide new insights into the molecular mechanisms underlying its pathogenicity. PMID- 21622756 TI - Quantitative analysis of spatial-temporal correlations during germination of spores of Bacillus Species. AB - Bacteria of Bacillus species sporulate upon starvation, and the resultant dormant spores germinate when the environment appears likely to allow the resumption of vegetative growth. Normally, the rates of germination of individual spores in populations are very heterogeneous, and the current work has investigated whether spore-to-spore communication enhances the synchronicity of germination. In order to do this work, time-lapse optical images of thousands of individual spores were captured during germination, and an image analysis algorithm was developed to do the following: (i) measure the positions and germination rates of many thousands of individual spores and (ii) compute pairwise correlations of their germination. This analysis showed that an individual spore's germination rate was dependent on its distance from other spores, especially at short distances. Thus, spores that were within a few micrometers exhibited an increased synchronicity in germination, suggesting that there is a mechanism for short-range communication between such spores during germination. However, two molecules known to be germinants that are released during germination, l-alanine and the 1:1 chelate of Ca2+ and dipicolinic acid, did not mediate spore-to-spore communication during germination. PMID- 21622757 TI - Functional organization of MobB, a small protein required for efficient conjugal transfer of plasmid R1162. AB - MobB is a small (molecular weight = 15,097) protein encoded by the broad-host range plasmid R1162 and is required for its efficient transfer by conjugation. The C-terminal half of the protein contains a membrane domain essential for transfer. This region can be replaced by a putative membrane domain from another, unrelated protein, and thus is likely to function independently from the rest of MobB. The other, functionally active region of MobB, identified by mutagenesis, is at the N-terminal end. One mutation affecting this region inhibits replication, suggesting that this part of the protein is contacting and sequestering the relaxase-linked primase. The overall organization reflects a multimeric and bipolar organization, with molecules of MobB anchored in the membrane at one end and engaging the relaxase at the other. This arrangement could increase the transfer frequency by raising the probability of contact between the relaxase and the membrane-embedded, coupling protein for type IV secretion. PMID- 21622758 TI - Draft genome sequence of Bacteroides vulgatus PC510, a strain isolated from human feces. AB - Although Bacteroides vulgatus is one of the most prevalent microorganisms in the human gastrointestinal tract, little is known about the genetic potential of this species. Here, we describe the annotated draft genome sequence of B. vulgatus PC510 isolated from human feces. PMID- 21622759 TI - Activity control of the ClpC adaptor McsB in Bacillus subtilis. AB - Controlled protein degradation is an important cellular reaction for the fast and efficient adaptation of bacteria to ever-changing environmental conditions. In the low-GC, Gram-positive model organism Bacillus subtilis, the AAA+ protein ClpC requires specific adaptor proteins not only for substrate recognition but also for chaperone activity. The McsB adaptor is activated particularly during heat stress, allowing the controlled degradation of the CtsR repressor by the ClpCP protease. Here we report how the McsB adaptor becomes activated by autophosphorylation on specific arginine residues during heat stress. In nonstressed cells McsB activity is inhibited by ClpC as well as YwlE. PMID- 21622760 TI - Communicating radiation risks to the public. PMID- 21622762 TI - Importance of rehabilitation in cancer treatment and palliative medicine. AB - Although rehabilitation for cancer patients is being practiced in clinical settings, it has not been very well recognized in cancer care. However, interest has been turning to cancer rehabilitation in recent years in association with advances in palliative care and the increasing numbers of patients who survive for long periods, while enduring symptoms caused by cancer or adverse effects associated with treatment. The fact that cancer patient rehabilitation fees were newly established in the 2010 revision of the Japanese medical service fees has propelled interest in this topic. Rehabilitation can be applied throughout the entire phase from the time of diagnosis to the terminal stage, and it is an approach that can involve psychosocial aspects as well as physical aspects. Although its effectiveness has not been adequately demonstrated, especially in the area of palliative medicine, rehabilitation for cancer patients is expected to be an important means of supporting the hopes of patients and their families, and attempting to maintain and improve patients' quality of life. PMID- 21622763 TI - A case of recurrent bile duct cancer initially treated with pancreaticoduodenectomy. PMID- 21622764 TI - Time trends in total cancer mortality (All Sites) between 1950 and 2008 in Japan, USA and Europe based on the WHO mortality database. PMID- 21622765 TI - Repair of bone erosions in rheumatoid arthritis treated with tumour necrosis factor inhibitors is based on bone apposition at the base of the erosion. AB - OBJECTIVES: To investigate whether bone erosions in patients with rheumatoid arthritis (RA) show evidence of repair. METHODS: 127 erosions were identified in metacarpophalangeal joints 2-4 of the right hands of 30 RA patients treated with tumour necrosis factor inhibitors (TNFi) and 21 sex, age and disease activity matched patients treated with methotrexate. All erosions were assessed for their exact maximal width and depth by high-resolution uCT imaging at baseline and after 1 year. RESULTS: All erosions detected at baseline could be visualised at follow-up after 1 year. At baseline, the mean width of bone erosions in the TNFi group was 2.0 mm; their mean depth was 2.3 mm, which was not significantly different from the methotrexate-treated group (width 2.4 mm; depth 2.4 mm). Mean depth of erosions significantly decreased after 1 year of treatment with TNFi ( 0.1 mm; p=0.016), whereas their width remained unchanged. In contrast, mean depth and width of erosive lesions increased in the methotrexate-treated group. The reduction in the depth of lesions was confined to erosions showing evidence of sclerosis at the base of the lesion. Moreover, deeper lesions in the TNFi group were particularly prone to repair (-0.4 mm; p=0.02) compared with more shallow lesions. CONCLUSIONS: Bone erosions in RA patients treated with TNFi show evidence of limited repair in contrast to bone erosions in patients treated with methotrexate. Repair is associated with a decrease in the depth of lesions and sclerosis at the bases of the lesions. Repair thus emerges from the endosteal rather than periosteal bone compartment and probably involves the bone marrow. PMID- 21622766 TI - Prevalence, incidence and progression of hand osteoarthritis in the general population: the Framingham Osteoarthritis Study. AB - OBJECTIVES: To describe the prevalence and longitudinal course of radiographic, erosive and symptomatic hand osteoarthritis (HOA) in the general population. METHODS: Framingham osteoarthritis (OA) study participants obtained bilateral hand radiographs at baseline and 9-year follow-up. The authors defined radiographic HOA at joint level as Kellgren-Lawrence grade (KLG)>=2, erosive HOA as KLG>=2 plus erosion and symptomatic HOA as KLG>=2 plus pain/aching/stiffness. Presence of HOA at individual level was defined as >=1 affected joint. The prevalence was age-standardised (US 2000 Population 40-84 years). RESULTS: Mean (SD) baseline age was 58.9 (9.9) years (56.5% women). The age-standardised prevalence of HOA was only modestly higher in women (44.2%) than men (37.7%), whereas the age-standardised prevalence of erosive and symptomatic OA was much higher in women (9.9% vs 3.3%, and 15.9% vs 8.2%). The crude incidence of HOA over 9-year follow-up was similar in women (34.6%) and men (33.7%), whereas the majority of those women (96.4%) and men (91.4%) with HOA at baseline showed progression during follow-up. Incident metacarpophalangeal and wrist OA were rare, but occurred more frequently and from an earlier age in men than women. Development of erosive disease occurred mainly in those with non-erosive HOA at baseline (as opposed to those without HOA), and was more frequent in women (17.3%) than men (9.6%). CONCLUSIONS: The usual female predominance of prevalent and incident HOA was less clear for radiographic HOA than for symptomatic and erosive HOA. With an ageing population, the impact of HOA will further increase. PMID- 21622767 TI - Serotyping for an extended anti-citrullinated peptide autoantibody panel does not add value to CCP2 testing for diagnosing RA in an early undifferentiated arthritis cohort. PMID- 21622768 TI - Adipokine profile is modulated in subcutaneous adipose tissue by TNFalpha inhibitors in patients with rheumatoid arthritis. PMID- 21622769 TI - Diagnostic performance of the ACR/EULAR 2010 criteria for rheumatoid arthritis and two diagnostic algorithms in an early arthritis clinic (REACH). AB - INTRODUCTION: An ACR/EULAR task force released new criteria to classify rheumatoid arthritis at an early stage. This study evaluates the diagnostic performance of these criteria and algorithms by van der Helm and Visser in REACH. METHODS: Patients with symptoms <=12 months from REACH were used. Algorithms were tested on discrimination, calibration and diagnostic accuracy of proposed cut points. Two patient sets were defined to test robustness; undifferentiated arthritis (UA) (n=231) and all patients including those without synovitis (n=513). The outcomes evaluated were methotrexate use and persistent disease at 12 months. RESULTS: In UA patients all algorithms had good areas under the curve 0.79, 95% CI 0.73 to 0.83 for the ACR/EULAR criteria, 0.80, 95% CI 0.74 to 0.87 for van der Helm and 0.83, 95% CI 0.77 to 0.88 for Visser. All calibrated well. Sensitivity and specificity were 0.74 and 0.66 for the ACR/EULAR criteria, 0.1 and 1.0 for van der Helm and 0.59 and 0.93 for Visser. Similar results were found in all patients indicating robustness. CONCLUSION: The ACR/EULAR 2010 criteria showed good diagnostic properties in an early arthritis cohort reflecting daily practice, as did the van der Helm and Visser algorithms. All were robust. To promote uniformity and comparability the ACR/EULAR 2010 criteria should be used in future diagnostic studies. PMID- 21622770 TI - Why should we expect a structure-modifying osteoarthritis drug to relieve osteoarthritis pain? PMID- 21622771 TI - Dysbalance of angiogenic and angiostatic mediators in patients with mixed connective tissue disease. AB - OBJECTIVE: Vascular disease is common in mixed connective tissue disease (MCTD). The aim of the present study was to investigate, whether dysbalance of angiogenic and angiostatic factors occurs in MCTD. METHODS: In all, 38 patients with MCTD, and 40 patients with systemic sclerosis (SSc) for comparison, were included. Four centres contributed to this cross-sectional analysis. A total of 66 healthy volunteers were used as controls. The serum levels of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and endostatin were determined by ELISA. For comparisons between controls and patients with MCTD and detection of associations of serum levels with dichotomous clinical parameters in patients with MCTD the Mann-Whitney test was used. RESULTS: Serum levels of the angiogenic factor VEGF were significantly elevated in patients with MCTD and SSc. Significantly increased levels of the angiostatic factor endostatin were also detected in MCTD, but not in SSc. No differences were observed for bFGF. Levels of VEGF were higher in patients with MCTD with pulmonary arterial hypertension (PAH), acrosclerosis and myositis. In multivariate linear regression analysis, an additive model of PAH, myositis and lymphadenopathy accounted for 79% of the variability of the VEGF levels (r=0.889). CONCLUSIONS: Molecular factors modulating angiogenic responses are dysregulated in patients with MCTD and SSc with increases of VEGF in MCTD and SSc and selective upregulation of endostatin in MCTD. Furthermore, high serum levels of VEGF might characterise patients with MCTD with a more severe course of the disease with increased prevalence of PAH and myositis. PMID- 21622772 TI - Clinical, radiographic, molecular and MRI-based predictors of cartilage loss in knee osteoarthritis. AB - OBJECTIVE: To examine the relationship of baseline clinical, radiographic, molecular and MRI measures with structural progression (subregional MRI-based femorotibial cartilage loss) in knee osteoarthritis (OA). METHODS: Single knees of 75 female participants with radiographic knee OA (and 77 healthy control participants) were examined over 24 months using MRI. Subregional femorotibial cartilage thickness was determined at baseline and follow-up. Baseline clinical, radiographic, molecular (n=16) and quantitative MRI-based measures of the meniscus and cartilage, including delayed gadolinium-enhanced MRI (dGEMRIC) and T2, were obtained. Differences in these baseline measures between radiographic osteoarthritic knees with longitudinal cartilage thinning (or thickening) and those with no significant change were evaluated by receiver operator characteristic analyses and Wilcoxon rank sum tests. RESULTS: The relatively strongest predictors of longitudinal cartilage thinning were reduced baseline cartilage thickness in the medial femur (area under the curve (AUC)=0.81), varus malalignment (AUC=0.77), reduced minimum joint space width and a greater radiographic joint space narrowing (JSN) score (both AUC=0.74). These remained significant after adjusting for multiple comparisons using false discovery rates. Reduced bone resorption (C-terminal telopeptide of type I collagen; AUC=0.65) and a low dGEMRIC index (reflecting low proteoglycan content) in the medial tibia (AUC=0.68) were associated with longitudinal cartilage thinning, but failed to reach statistical significance after correction for multiple testing in this (small) sample. CONCLUSIONS: This exploratory study indicates that baseline molecular or MRI cartilage compositional markers may not provide better discrimination between knees with cartilage thinning and those without longitudinal change than simple radiographic measures, such as greater JSN score. PMID- 21622773 TI - Interrater reliability and aspects of validity of the myositis damage index. AB - OBJECTIVE: To test the interrater reliability, internal consistency and aspects of validity of the myositis damage index (MDI) in the assessment of damage in adult patients with idiopathic inflammatory myopathy (IIM). METHODS: 95 patients were assessed in six centres as part of this cross-sectional international study. Two parts of a MDI were used to assess disease damage, the MDI and the myositis damage score (MYODAM). The myositis disease activity assessment tool (MDAAT) was used to assess disease activity. Interrater reliability was assessed using intraclass correlation coefficient (ICC). Spearman's rank correlation coefficient was used to measure the convergent validity of cross-sectional scores between the two parts of the damage tool and to determine the correlation between the respective components of the damage and activity tools. RESULTS: In general, the damage index appears to have good interrater reliability for most of the systems with an ICC greater than 0.65. Convergent validity between the two parts of the damage tool showed good correlation for the individual organ systems (r>0.8). There were weak correlations between some parts of the MDI and corresponding components of the MDAAT. CONCLUSION: The MDI is a comprehensive tool to assess damage in patients with myositis. With physician education and emphasis to record items that have been diagnosed since the myositis diagnosis, the MDI will provide a valuable tool to assess damage in future clinical trials and longitudinal studies. PMID- 21622774 TI - Validation of pain and patient global scales in chronic gout: data from two randomised controlled trials. AB - OBJECTIVE: To assess validity of pain and patient global scales in gout. METHODS: The authors used data from pegloticase clinical trials in chronic refractory gout to examine the validity of visual analogue scale (VAS) pain, Short-Form 36 (SF 36) bodily pain subscale and VAS patient global assessment (all scales 0-100). Convergent/divergent validity with clinical characteristics was tested by using Spearman's correlation coefficient. For discriminant ability, the authors compared the change at 6 months between placebo and pegloticase arms and calculated effect size (ES) and standardised response mean (SRM). RESULTS: 212 patients (mean age, 55.4 years, 82% men; 73% with tophaceous gout) provided data. VAS pain was statistically significantly correlated with tender joints (r=0.42), swollen joints (r=0.30), SF-36 physical (r=-0.56) and Mental Component Summary (r=-0.36) and Health Assessment Questionnaire scores (r=0.54; all p-values <0.0001), but not disease duration (r=-0.01; p=0.84), gout flares (r=0.12; p=0.08), comorbidities (r=0.05; p=0.47) or plasma urate (r=0.01; p=0.89). Similar and significant correlation coefficients with tender and swollen joints were noted for VAS patient global assessment (r=0.35 and 0.23; p<0.0012 for both) and SF-36 pain subscale (r=-0.27 and -0.19; p<0.006 for both). Pegloticase group had significantly more improvement than placebo at 6 months, mean (SD): VAS pain, 9.2 (29.3) versus 1.9 (26.4), p=0.0002; SF-36 pain, 14.6 (25.6) versus -0.04 (21.1), p<0.0001; and patient global, -9.3 (26.5) versus 3.4 (22.8), p<0.0001. ES and SRMs in pegloticase group were as follows: VAS pain, 0.34 and 0.30; SF-36 pain, 0.69 and 0.57; patient global, 0.49 and 0.44. CONCLUSION: VAS pain, SF-36 pain and patient global VAS are valid outcome measures in patients with chronic gout. PMID- 21622775 TI - Efficacy of allogeneic mesenchymal stem cell transplantation in patients with drug-resistant polymyositis and dermatomyositis. AB - OBJECTIVE: To assess the safety and clinical efficacy of allogeneic mesenchymal stem cell transplantation (MSCT) in a small-scale pilot study with 10 patients with drug-resistant polymyositis (PM) or dermatomyositis (DM). METHODS: A single arm trial involving 10 patients with DM/PM who were either refractory to standard treatment, or had severe systemic involvement. All patients consented and underwent allogeneic MSCT. Clinical and laboratory manifestations were compared before and after MSCT. RESULTS: Improvements were seen in serum creatine kinase (CK), CK-MB, patient global assessment by visual analogue scale and muscle strength by manual muscle test in all patients, as well as improvement in interstitial lung disease in selected patients. Improvement in chronic non healing skin ulcers was noted in one patient. Clinical responses were also seen in patients undergoing a second MSCT for recurrence of disease. CONCLUSION: MSCT appears safe and effective in drug-resistant patients with DM/PM. Larger-scale studies including a control group receiving standard treatment are needed to assess the long-term efficacy of allogeneic MSCT in refractory patients with DM/PM. PMID- 21622776 TI - Genetic association of PRDM1-ATG5 intergenic region and autophagy with systemic lupus erythematosus in a Chinese population. AB - OBJECTIVE: Recent genome-wide association studies suggested the PRDM1-ATG5 gene region as a systemic lupus erythematosus (SLE)-associated locus both in Caucasian and Chinese populations; however, the candidate gene was still obscure and the possible functional significance needed to be determined. METHODS: In this study, by a multistage integrative strategy, the authors first performed a case-control association study involving 1745 individuals in the Chinese population by genotyping nine single nucleotide polymorphisms within this region, and a meta analysis was conducted. Correlation between associated genotypes and expression levels of messenger RNA in B-cell lines from 210 unrelated HapMap data was examined, and was validated in vitro. To determine the biological significance, a genetic association study was also checked in a pathway-based manner and the significant associations were validated in a second 844 Chinese cohort. RESULTS: A peak of association was found in the intergenic region (p=0.036-3.26*10(-4)). Meta-analysis consolidated the association between rs548234 and SLE (OR 1.254, p=1.28*10(-16)). Significant positive correlations with ATG5 expression were identified, suggesting ATG5 as a candidate gene in the region. Epstein-Barr virus B-cell-based downstream gene expression analysis supported a functional effect of rs548234 and rs6937876, and in-vitro experiments confirmed the regulatory effect of rs6937876 in B-cell populations. Finally, an autophagy pathway-based genetic association study identified ATG7 (p=1.12*10(-4)) and IRGM (p=0.015) as novel candidate genes, and gene-gene interactions were observed between ATG5, ATG7 and IRGM. CONCLUSION: These data may demonstrate that autophagy is involved in the pathogenesis of SLE and imply a common biological pathway in autoimmunity. PMID- 21622777 TI - Local adaptation to soil hypoxia determines the structure of an arbuscular mycorrhizal fungal community in roots from natural CO2 springs. AB - The processes responsible for producing and maintaining the diversity of natural arbuscular mycorrhizal (AM) fungal communities remain largely unknown. We used natural CO(2) springs (mofettes), which create hypoxic soil environments, to determine whether a long-term, directional, abiotic selection pressure could change AM fungal community structure and drive the selection of particular AM fungal phylotypes. We explored whether those phylotypes that appear exclusively in hypoxic soils are local specialists or widespread generalists able to tolerate a range of soil conditions. AM fungal community composition was characterized by cloning, restriction fragment length polymorphism typing, and the sequencing of small subunit rRNA genes from roots of four plant species growing at high (hypoxic) and low (control) geological CO(2) exposure. We found significant levels of AM fungal community turnover (beta diversity) between soil types and the numerical dominance of two AM fungal phylotypes in hypoxic soils. Our results strongly suggest that direct environmental selection acting on AM fungi is a major factor regulating AM fungal communities and their phylogeographic patterns. Consequently, some AM fungi are more strongly associated with local variations in the soil environment than with their host plant's distribution. PMID- 21622778 TI - Detection of known and novel adenoviruses in cattle wastes via broad-spectrum primers. AB - The critical assessment of bovine adenoviruses (BAdV) as indicators of environmental fecal contamination requires improved knowledge of their prevalence, shedding dynamics, and genetic diversity. We examined DNA extracted from bovine and other animal waste samples collected in Wisconsin for atadenoviruses and mastadenoviruses using novel, broad-spectrum PCR primer sets. BAdV were detected in 13% of cattle fecal samples, 90% of cattle urine samples, and 100% of cattle manure samples; 44 percent of BAdV-positive samples contained both Atadenovirus and Mastadenovirus DNA. Additionally, BAdV were detected in soil, runoff water from a cattle feedlot, and residential well water. Overall, we detected 8 of 11 prototype BAdV, plus bovine, rabbit, and porcine mastadenoviruses that diverged significantly from previously reported genotypes. The prevalence of BAdV shedding by cattle supports targeting AdV broadly as indicators of the presence of fecal contamination in aqueous environments. Conversely, several factors complicate the use of AdV for fecal source attribution. Animal AdV infecting a given livestock host were not monophyletic, recombination among livestock mastadenoviruses was detected, and the genetic diversity of animal AdV is still underreported. These caveats highlight the need for continuing genetic surveillance for animal AdV and for supporting data when BAdV detection is invoked for fecal source attribution in environmental samples. To our knowledge, this is the first study to report natural BAdV excretion in urine, BAdV detection in groundwater, and recombination in AdV of livestock origin. PMID- 21622779 TI - Generalized relationship between numbers of bacteria and their viability in biofilms. AB - Bacterial biofilms are confined communities that are encapsulated in protective layers of extracellular polymeric substances. Microscopic evaluation of biofilms of diverse bacterial strains on various substrata reveals that, in general, the percentage of viable bacteria decreases with the total number of bacteria in a biofilm. PMID- 21622780 TI - Evaluation of physical coverings used to control Escherichia coli O157:H7 at the compost heap surface. AB - Throughout four field trials, compost heaps covered with finished compost maintained temperatures under the physical covering that were ca. 7 to 15.5 degrees C higher, resulting in rapid Escherichia coli O157:H7 reduction, than those of the heaps covered with fresh straw or left uncovered. Our results validated recommendations made by the U.S. Environmental Protection Agency for covering fresh compost. PMID- 21622781 TI - Methylotrophy in a lake: from metagenomics to single-organism physiology. AB - This review provides a brief summary of ongoing studies in Lake Washington (Seattle, WA) directed at an understanding of the content and activities of microbial communities involved in methylotrophy. One of the findings from culture independent approaches, including functional metagenomics, is the prominent presence of Methylotenera species in the site and their inferred activity in C(1) metabolism, highlighting the local environmental importance of this group. Comparative analyses of individual genomes of Methylophilaceae from Lake Washington provide insights into their genomic divergence and suggest significant metabolic flexibility. PMID- 21622782 TI - Enhanced removal of a human norovirus surrogate from fresh vegetables and fruits by a combination of surfactants and sanitizers. AB - Fruits and vegetables are major vehicles for transmission of food-borne enteric viruses since they are easily contaminated at pre- and postharvest stages and they undergo little or no processing. However, commonly used sanitizers are relatively ineffective for removing human norovirus surrogates from fresh produce. In this study, we systematically evaluated the effectiveness of surfactants on removal of a human norovirus surrogate, murine norovirus 1 (MNV 1), from fresh produce. We showed that a panel of surfactants, including sodium dodecyl sulfate (SDS), Nonidet P-40 (NP-40), Triton X-100, and polysorbates, significantly enhanced the removal of viruses from fresh fruits and vegetables. While tap water alone and chlorine solution (200 ppm) gave only <1.2-log reductions in virus titer in all fresh produce, a solution containing 50 ppm of surfactant was able to achieve a 3-log reduction in virus titer in strawberries and an approximately 2-log reduction in virus titer in lettuce, cabbage, and raspberries. Moreover, a reduction of approximately 3 logs was observed in all the tested fresh produce after sanitization with a solution containing a combination of 50 ppm of each surfactant and 200 ppm of chlorine. Taken together, our results demonstrate that the combination of a surfactant with a commonly used sanitizer enhanced the efficiency in removing viruses from fresh produce by approximately 100 times. Since SDS is an FDA-approved food additive and polysorbates are recognized by the FDA as GRAS (generally recognized as safe) products, implementation of this novel sanitization strategy would be a feasible approach for efficient reduction of the virus load in fresh produce. PMID- 21622783 TI - Persistence of avian influenza viruses in lake sediment, duck feces, and duck meat. AB - The persistence of 3 low-pathogenicity avian influenza viruses (LPAIV) (H4N6, H5N1, and H6N8) and one human influenza virus (H1N1) as well as Newcastle disease virus (NDV) and enteric cytopathogenic bovine orphan (ECBO) virus was investigated in lake sediment, duck feces, and duck meat at 30, 20, 10, and 0 degrees C using a germ carrier technique. Virus-loaded germ carriers were incubated in each substrate, and residual infectivity of the eluted virus was quantified on cell culture after regular intervals for a maximum of 24 weeks. Data were analyzed by a linear regression model to calculate T(90) values (time required for 90% loss of virus infectivity) and estimated persistence of the viruses. In general, the persistence of all of the viruses was highest in lake sediment, followed by feces, and was the lowest in duck meat at all temperatures. For the avian influenza virus subtypes, T(90) values in sediment ranged from 5 to 11, 13 to 18, 43 to 54, and 66 to 394 days at 30, 20, 10, and 0 degrees C, respectively, which were 2 to 5 times higher than the T(90) values of the viruses in the feces and meat. Although the individual viruses vary in tenacity, the survival time of influenza viruses was shorter than that of NDV and ECBO virus in all substrates. The results of this study suggest that lake sediment may act as a long-term source of influenza viruses in the aquatic habitat, while the viruses may remain infectious for extended periods of time in duck feces and meat at low temperatures, allowing persistence of the viruses in the environment over winter. PMID- 21622784 TI - Impact of internal RNA on aggregation and electrokinetics of viruses: comparison between MS2 phage and corresponding virus-like particles. AB - We compare for the first time the electrokinetic and aggregation properties of MS2 phage (pH 2.5 to 7, 1 to 100 mM NaNO(3) electrolyte concentration) with those of the corresponding virus-like particles (VLPs), which lack entirely the inner viral RNA component. In line with our previous work (J. Langlet, F. Gaboriaud, C. Gantzer, and J. F. L. Duval, Biophys. J. 94:3293-3312, 2008), it is found that modifying the content of RNA within the virus leads to very distinct electrohydrodynamic and aggregation profiles for MS2 and MS2 VLPs. Under the given pH and concentration conditions, MS2 VLPs exhibit electrophoretic mobility larger in magnitude than that of MS2, and both have similar isoelectric point (IEP) values (~4). The electrokinetic results reflect a greater permeability of MS2 VLPs to electroosmotic flow, developed within/around these soft particles during their migration under the action of the applied electrical field. Results also support the presence of some remaining negatively charged component within the VLPs. In addition, MS2 phage systematically forms aggregates at pH values below the IEP, regardless of the magnitude of the solution ionic strength, whereas MS2 VLPs aggregate under the strict condition where the pH is relatively equal to the IEP at sufficiently low salt concentrations (<10 mM). It is argued that the stability of VLPs against aggregation and the differences between electrokinetics of MS2 and corresponding VLPs conform to recently developed formalisms for the stability and electrohydrodynamics of soft multilayered particles. The differences between the surface properties of these two kinds of particles reported here suggest that VLPs may not be appropriate for predicting the behavior of pathogenic viruses in aqueous media. PMID- 21622785 TI - Molecular detection of Campylobacter spp. in California gull (Larus californicus) excreta. AB - We examined the prevalence, quantity, and diversity of Campylobacter species in the excreta of 159 California gull (Larus californicus) samples using culture-, PCR-, and quantitative PCR (qPCR)-based detection assays. Campylobacter prevalence and abundance were relatively high in the gull excreta examined; however, C. jejuni and C. lari were detected in fewer than 2% of the isolates and DNA extracts from the fecal samples that tested positive. Moreover, molecular and sequencing data indicated that most L. californicus campylobacters were novel (<97% 16S rRNA gene sequence identity to known Campylobacter species) and not closely related to species commonly associated with human illness. Campylobacter estimates were positively related with those of fecal indicators, including a gull fecal marker based on the Catellicoccus marimammalium 16S rRNA gene. PMID- 21622786 TI - Impact of biochar application to soil on the root-associated bacterial community structure of fully developed greenhouse pepper plants. AB - Adding biochar to soil has environmental and agricultural potential due to its long-term carbon sequestration capacity and its ability to improve crop productivity. Recent studies have demonstrated that soil-applied biochar promotes the systemic resistance of plants to several prominent foliar pathogens. One potential mechanism for this phenomenon is root-associated microbial elicitors whose presence is somehow augmented in the biochar-amended soils. The objective of this study was to assess the effect of biochar amendment on the root associated bacterial community composition of mature sweet pepper (Capsicum annuum L.) plants. Molecular fingerprinting (denaturing gradient gel electrophoresis and terminal restriction fragment length polymorphism) of 16S rRNA gene fragments showed a clear differentiation between the root-associated bacterial community structures of biochar-amended and control plants. The pyrosequencing of 16S rRNA amplicons from the rhizoplane of both treatments generated a total of 20,142 sequences, 92 to 95% of which were affiliated with the Proteobacteria, Bacteroidetes, Actinobacteria, and Firmicutes phyla. The relative abundance of members of the Bacteroidetes phylum increased from 12 to 30% as a result of biochar amendment, while that of the Proteobacteria decreased from 71 to 47%. The Bacteroidetes-affiliated Flavobacterium was the strongest biochar-induced genus. The relative abundance of this group increased from 4.2% of total root-associated operational taxonomic units (OTUs) in control samples to 19.6% in biochar-amended samples. Additional biochar-induced genera included chitin and cellulose degraders (Chitinophaga and Cellvibrio, respectively) and aromatic compound degraders (Hydrogenophaga and Dechloromonas). We hypothesize that these biochar-augmented genera may be at least partially responsible for the beneficial effect of biochar amendment on plant growth and viability. PMID- 21622787 TI - Cultivation-independent detection of autotrophic hydrogen-oxidizing bacteria by DNA stable-isotope probing. AB - Knallgas bacteria are a physiologically defined group that is primarily studied using cultivation-dependent techniques. Given that current cultivation techniques fail to grow most bacteria, cultivation-independent techniques that selectively detect and identify knallgas bacteria will improve our ability to study their diversity and distribution. We used stable-isotope probing (SIP) to identify knallgas bacteria in rhizosphere soil of legumes and in a microbial mat from Obsidian Pool in Yellowstone National Park. When samples were incubated in the dark, incorporation of (13)CO(2) was H(2) dependent. SIP enabled the detection of knallgas bacteria that were not detected by cultivation, and the majority of bacteria identified in the rhizosphere soils were betaproteobacteria predominantly related to genera previously known to oxidize hydrogen. Bacteria in soil grew on hydrogen at concentrations as low as 100 ppm. A hydB homolog encoding a putative high-affinity NiFe hydrogenase was amplified from (13)C labeled DNA from both vetch and clover rhizosphere soil. The results indicate that knallgas bacteria can be detected by SIP and populations that respond to different H(2) concentrations can be distinguished. The methods described here should be applicable to a variety of ecosystems and will enable the discovery of additional knallgas bacteria that are resistant to cultivation. PMID- 21622788 TI - Rapid fluorescence-based screening for Wolbachia endosymbionts in Drosophila germ line and somatic tissues. AB - Wolbachia is a globally distributed bacterial endosymbiont present in arthropods and nematodes. The advent of sensitive PCR-based approaches has greatly facilitated the identification of Wolbachia-infected individuals and analysis of population infection levels. Here, a complementary visual fluorescence-based Wolbachia screening approach is described. Through the use of the fluorescent dye Syto-11, Wolbachia can be efficiently detected in various Drosophila tissues, including ovaries. Syto-11 also stains Wolbachia in other insects. Because Wolbachia is inherited through the maternal germ line, bacteria reside in the ovaries of flies in infected populations. An advantage of this staining approach is that it informs about Wolbachia titer as well as its tissue and cellular distribution. Using this method, the infection status of insect populations in two central California locations was determined, and variants with unusually low or high Wolbachia titers were isolated. In addition, a variant with ovarioles containing both infected and uninfected egg chambers was identified. Syto-11 staining of Cardinium- and Spiroplasma-infected insects was also analyzed. PMID- 21622789 TI - SidL, an Aspergillus fumigatus transacetylase involved in biosynthesis of the siderophores ferricrocin and hydroxyferricrocin. AB - The opportunistic fungal pathogen Aspergillus fumigatus produces four types of siderophores, low-molecular-mass iron chelators: it excretes fusarinine C (FsC) and triacetylfusarinine C (TAFC) for iron uptake and accumulates ferricrocin (FC) for hyphal and hydroxyferricrocin (HFC) for conidial iron distribution and storage. Siderophore biosynthesis has recently been shown to be crucial for fungal virulence. Here we identified a new component of the fungal siderophore biosynthetic machinery: AFUA_1G04450, termed SidL. SidL is conserved only in siderophore-producing ascomycetes and shows similarity to transacylases involved in bacterial siderophore biosynthesis and the N(5) hydroxyornithine:anhydromevalonyl coenzyme A-N(5)-transacylase SidF, which is essential for TAFC biosynthesis. Inactivation of SidL in A. fumigatus decreased FC biosynthesis during iron starvation and completely blocked FC biosynthesis during iron-replete growth. In agreement with these findings, SidL deficiency blocked conidial accumulation of FC-derived HFC under iron-replete conditions, which delayed germination and decreased the size of conidia and their resistance to oxidative stress. Remarkably, the sidL gene is not clustered with other siderophore-biosynthetic genes, and its expression is not affected by iron availability. Tagging of SidL with enhanced green fluorescent protein suggested a cytosolic localization of the FC-biosynthetic machinery. Taken together, these data suggest that SidL is a constitutively active N(5)-hydroxyornithine-acetylase required for FC biosynthesis, in particular under iron-replete conditions. Moreover, this study revealed the unexpected complexity of siderophore biosynthesis, indicating the existence of an additional, iron-repressed N(5) hydroxyornithine-acetylase. PMID- 21622790 TI - Different IS629 transposition frequencies exhibited by Escherichia coli O157:H7 strains in the stepwise evolutionary model. AB - The insertion sequence IS629, which is highly prevalent in Escherichia coli O157:H7 genomes, was found to be absent in O157:H- strains, which are on a divergent pathway in the emergence of O157:H7. Although O157:H- is deficient in IS629, it permits IS629 transposition, with an excision frequency higher than that of ancestral O55:H7 strains but significantly lower than that of pathogenic O157:H7 strains. PMID- 21622791 TI - Development of a multilocus sequence typing tool for high-resolution genotyping of Enterocytozoon bieneusi. AB - Thus far, genotyping of Enterocytozoon bieneusi has been based solely on DNA sequence analysis of the internal transcribed spacer (ITS) of the rRNA gene. Both host-adapted and zoonotic (human-pathogenic) genotypes of E. bieneusi have been identified. In this study, we searched for microsatellite and minisatellite sequences in the whole-genome sequence database of E. bieneusi isolate H348. Seven potential targets (MS1 to MS7) were identified. Testing of the seven targets by PCR using two human-pathogenic E. bieneusi genotypes (A and Peru10) led to the selection of four targets (MS1, MS3, MS4, and MS7). Further analysis of the four loci with an additional 24 specimens of both host-adapted and zoonotic E. bieneusi genotypes indicated that most host-adapted genotypes were not amplified by PCR targeting these loci. In contrast, 10 or 11 of the 13 specimens of the zoonotic genotypes were amplified by PCR at each locus. Altogether, 12, 8, 7, and 11 genotypes of were identified at MS1, MS3, MS4, and MS7, respectively. Phylogenetic analysis of the nucleotide sequences obtained produced a genetic relationship that was similar to the one at the ITS locus, with the formation of a large group of zoonotic genotypes that included most E. bieneusi genotypes in humans. Thus, a multilocus sequence typing tool was developed for high-resolution genotyping of E. bieneusi. Data obtained in the study should also have implications for understanding the taxonomy of Enterocytozoon spp., the public health significance of E. bieneusi in animals, and the sources of human E. bieneusi infections. PMID- 21622792 TI - Comparison of genotypic and phylogenetic relationships of environmental Enterococcus isolates by BOX-PCR typing and 16S rRNA gene sequencing. AB - Environmental Enterococcus spp. were compared by BOX-PCR genotyping and 16S rRNA gene sequencing to clarify the predictive relationship of BOX-PCR fingerprints to species designation. BOX-PCR and 16S rRNA gene relationships agreed for 77% of strains. BOX-PCR provided superior intraspecies discrimination but incorrectly identified some strains to the species level and divided some species into multiple groups. PMID- 21622793 TI - Cell surface display of poliovirus receptor on Escherichia coli, a novel method for concentrating viral particles in water. AB - The lack of efficient methods for concentrating viruses in water samples leads to underreporting of viral contamination in source water. A novel strategy for viral concentration was developed using the expression of target virus receptors on bacterial cells. Poliovirus type 1, the most studied enterovirus, was used as a surrogate for enteric viruses. The human poliovirus receptor (hPVR) gene was expressed on the surface of Escherichia coli cells by using the ice nucleation protein (INP) gene. The hPVR gene was ligated to the 3' end of the INP gene after the removal of the stop codon. The resulting open reading frame (ORF) was used for the projection of hPVR onto the outer membrane of E. coli. Gene expression was tested by SDS-PAGE, Western blot, and dot blot analyses, and virion capture ability was confirmed by transmission electron microscopy. The application of engineered E. coli cells for capturing viruses in 1-liter samples of source and drinking water resulted in 75 to 99% procedural recovery efficiency. Cell surface display of viral receptors on bacterial cells opens a new prospect for an efficient and inexpensive alternative tool for capturing and concentrating waterborne viruses in water samples. PMID- 21622794 TI - Restructuring of endophytic bacterial communities in grapevine yellows-diseased and recovered Vitis vinifera L. plants. AB - Length heterogeneity-PCR assays, combined with statistical analyses, highlighted that the endophytic bacterial community associated with healthy grapevines was characterized by a greater diversity than that present in diseased and recovered plants. The findings suggest that phytoplasmas can restructure the bacterial community by selecting endophytic strains that could elicit a plant defense response. PMID- 21622795 TI - Role played by exosporium glycoproteins in the surface properties of Bacillus cereus spores and in their adhesion to stainless steel. AB - Bacillus cereus spores are surrounded by a loose-fitting layer called the exosporium, whose distal part is mainly formed from glycoproteins. The role played by the exosporium glycoproteins of B. cereus ATCC 14579 (BclA and ExsH) was investigated by considering hydrophobicity and charge, as well as the properties of spore adhesion to stainless steel. The absence of BclA increased both the isoelectric point (IEP) and hydrophobicity of whole spores while simultaneously reducing the interaction between spores and stainless steel. However, neither the hydrophobicity nor the charge associated with BclA could explain the differences in the adhesion properties. Conversely, ExsH, another exosporium glycoprotein, did not play a significant role in spore surface properties. The monosaccharide analysis of B. cereus ATCC 14579 showed different glycosylation patterns on ExsH and BclA. Moreover, two specific glycosyl residues, namely, 2-O-methyl-rhamnose (2-Me-Rha) and 2,4-O-methyl-rhamnose (2,4 Me-Rha), were attached to BclA, in addition to the glycosyl residues already reported in B. anthracis. PMID- 21622796 TI - Cholesterol degradation by Gordonia cholesterolivorans. AB - This paper reports physiological and genetic data about the type strain Gordonia cholesterolivorans, a strain that is able to degrade steroid compounds containing a long carbon side chain such as cholesterol (C(27)), cholestenone (C(27)), ergosterol (C(28)), and stigmasterol (C(29)). The length of the carbon side chain appears to be of great importance for this bacterium, as the strain is unable to grow using steroids with a shorter or nonaliphatic carbon side chain such as cholic acid (C(24)), progesterone (C(21)), testosterone, androsterone, 4 androstene-3,17-dione (all C(19)), and further steroids. This study also demonstrates that the degradation of cholesterol is a quite common feature of the genus Gordonia by comparing Gordonia cholesterolivorans with some other species of this genus (e.g., G. sihwensis, G. hydrophobica, G. australis, and G. neofelifaecis). Pyrosequencing of the genome of G. cholesterolivorans led to the identification of two conventional cholesterol oxidase genes on an 8-kb and a 12.8-kb genomic fragment with genetic organizations that are quite unique as compared to the genomes of other cholesterol-degrading bacteria sequenced so far. The identified two putative cholesterol oxidases of G. cholesterolivorans are both intracellularly acting enzymes of the class I type. Whereas one of these two cholesterol oxidases (ChoOx-1) shows high identity with an oxidoreductase of the opportunistic pathogen G. bronchialis and is not transcribed during growth with cholesterol, the other one (ChoOx-2) appears phylogenetically closer to cholesterol oxidases from members of the genus Rhodococcus and is transcribed constitutively. By using targeted gene disruption, a G. cholesterolivorans ChoOx 2 gene mutant strain that was unable to grow with steroids was obtained. PMID- 21622797 TI - Assembly of minicellulosomes on the surface of Bacillus subtilis. AB - To cost-efficiently produce biofuels, new methods are needed to convert lignocellulosic biomass into fermentable sugars. One promising approach is to degrade biomass using cellulosomes, which are surface-displayed multicellulase containing complexes present in cellulolytic Clostridium and Ruminococcus species. In this study we created cellulolytic strains of Bacillus subtilis that display one or more cellulase enzymes. Proteins containing the appropriate cell wall sorting signal are covalently anchored to the peptidoglycan by coexpressing them with the Bacillus anthracis sortase A (SrtA) transpeptidase. This approach was used to covalently attach the Cel8A endoglucanase from Clostridium thermocellum to the cell wall. In addition, a Cel8A-dockerin fusion protein was anchored on the surface of B. subtilis via noncovalent interactions with a cell wall-attached cohesin module. We also demonstrate that it is possible to assemble multienzyme complexes on the cell surface. A three-enzyme-containing minicellulosome was displayed on the cell surface; it consisted of a cell wall attached scaffoldin protein noncovalently bound to three cellulase-dockerin fusion proteins that were produced in Escherichia coli. B. subtilis has a robust genetic system and is currently used in a wide range of industrial processes. Thus, grafting larger, more elaborate minicellulosomes onto the surface of B. subtilis may yield cellulolytic bacteria with increased potency that can be used to degrade biomass. PMID- 21622798 TI - Control of bacterial persister cells by Trp/Arg-containing antimicrobial peptides. AB - Persister cells are dormant phenotypic variants inherent in a bacterial population. They play important roles in chronic infections and present great challenges to therapy due to extremely enhanced tolerance to antibiotics compared to that of normal cells of the same genotype. In this study, we report that cationic membrane-penetrating peptides containing various numbers of arginine and tryptophan repeats are effective in killing persister cells of Escherichia coli HM22, a hyper-persister producer. The activities of three linear peptides [(RW)(n)-NH(2), where n is 2, 3, or 4] and a dendrimeric peptide, (RW)(4D), in killing bacterial persisters were compared. Although the dendrimeric peptide (RW)(4D) requires a lower threshold to kill planktonic persisters, octameric peptide (RW)(4)-NH(2) is the most effective against planktonic persister cells at high concentrations. For example, treatment with 80 MUM (RW)(4)-NH(2) for 60 min led to a 99.7% reduction in the number of viable persister cells. The viability of persister cells residing in surface-attached biofilms was also significantly reduced by (RW)(4)-NH(2) and (RW)(4D). These two peptides were also found to significantly enhance the susceptibility of biofilm cells to ofloxacin. The potency of (RW)(4)-NH(2) was further marked by its ability to disperse and kill preformed biofilms harboring high percentages of persister cells. Interestingly, approximately 70% of the dispersed cells were found to have lost their intrinsic tolerance and become susceptible to ampicillin if not killed directly by this peptide. These results are helpful for better understanding the activities of these peptides and may aid in future development of more effective therapies of chronic infections. PMID- 21622799 TI - Diversity, abundance, and potential activity of nitrifying and nitrate-reducing microbial assemblages in a subglacial ecosystem. AB - Subglacial sediments sampled from beneath Robertson Glacier (RG), Alberta, Canada, were shown to harbor diverse assemblages of potential nitrifiers, nitrate reducers, and diazotrophs, as assessed by amoA, narG, and nifH gene biomarker diversity. Although archaeal amoA genes were detected, they were less abundant and less diverse than bacterial amoA, suggesting that bacteria are the predominant nitrifiers in RG sediments. Maximum nitrification and nitrate reduction rates in microcosms incubated at 4 degrees C were 280 and 18.5 nmol of N per g of dry weight sediment per day, respectively, indicating the potential for these processes to occur in situ. Geochemical analyses of subglacial sediment pore waters and bulk subglacial meltwaters revealed low concentrations of inorganic and organic nitrogen compounds. These data, when coupled with a C/N atomic ratio of dissolved organic matter in subglacial pore waters of ~210, indicate that the sediment communities are N limited. This may reflect the combined biological activities of organic N mineralization, nitrification, and nitrate reduction. Despite evidence of N limitation and the detection of nifH, we were unable to detect biological nitrogen fixation activity in subglacial sediments. Collectively, the results presented here suggest a role for nitrification and nitrate reduction in sustaining microbial life in subglacial environments. Considering that ice currently covers 11% of the terrestrial landmass and has covered significantly greater portions of Earth at times in the past, the demonstration of nitrification and nitrate reduction in subglacial environments furthers our understanding of the potential for these environments to contribute to global biogeochemical cycles on glacial-interglacial timescales. PMID- 21622800 TI - Saccharomyces cerevisiae genome shuffling through recursive population mating leads to improved tolerance to spent sulfite liquor. AB - Spent sulfite liquor (SSL) is a waste effluent from sulfite pulping that contains monomeric sugars which can be fermented to ethanol. However, fermentative yeasts used for the fermentation of the sugars in SSL are adversely affected by the inhibitory substances in this complex feedstock. To overcome this limitation, evolutionary engineering of Saccharomyces cerevisiae was carried out using genome shuffling technology based on large-scale population cross mating. Populations of UV-light-induced yeast mutants more tolerant than the wild type to hardwood spent sulfite liquor (HWSSL) were first isolated and then recursively mated and enriched for more-tolerant populations. After five rounds of genome shuffling, three strains were isolated that were able to grow on undiluted HWSSL and to support efficient ethanol production from the sugars therein for prolonged fermentation of HWSSL. Analyses showed that greater HWSSL tolerance is associated with improved viability in the presence of salt, sorbitol, peroxide, and acetic acid. Our results showed that evolutionary engineering through genome shuffling will yield robust yeasts capable of fermenting the sugars present in HWSSL, which is a complex substrate containing multiple sources of inhibitors. These strains may not be obtainable through classical evolutionary engineering and can serve as a model for further understanding of the mechanism behind simultaneous tolerance to multiple inhibitors. PMID- 21622801 TI - Frequent gene movement and pseudogene evolution is common to the large and complex genomes of wheat, barley, and their relatives. AB - All six arms of the group 1 chromosomes of hexaploid wheat (Triticum aestivum) were sequenced with Roche/454 to 1.3- to 2.2-fold coverage and compared with similar data sets from the homoeologous chromosome 1H of barley (Hordeum vulgare). Six to ten thousand gene sequences were sampled per chromosome. These were classified into genes that have their closest homologs in the Triticeae group 1 syntenic region in Brachypodium, rice (Oryza sativa), and/or sorghum (Sorghum bicolor) and genes that have their homologs elsewhere in these model grass genomes. Although the number of syntenic genes was similar between the homologous groups, the amount of nonsyntenic genes was found to be extremely diverse between wheat and barley and even between wheat subgenomes. Besides a small core group of genes that are nonsyntenic in other grasses but conserved among Triticeae, we found thousands of genic sequences that are specific to chromosomes of one single species or subgenome. By examining in detail 50 genes from chromosome 1H for which BAC sequences were available, we found that many represent pseudogenes that resulted from transposable element activity and double strand break repair. Thus, Triticeae seem to accumulate nonsyntenic genes frequently. Since many of them are likely to be pseudogenes, total gene numbers in Triticeae are prone to pronounced overestimates. PMID- 21622802 TI - Assessment of nasal fibroscopy to explore olfactory cleft. AB - OBJECTIVE: To assess whether flexible nasoendoscopy can be used to visualize all parts of the olfactory cleft (OC) without morbidity. STUDY DESIGN: Single-center, prospective, observational study. SETTING: French tertiary referral center. SUBJECTS AND METHODS: 100 consecutive patients were divided in 2 groups of 50. Group 1 underwent nasal fibroscopy without vasoconstriction or local anesthesia with an endosheath- protected endoscope. Group 2 was examined by a fiberscope without an endosheath after application of naphazoline Xylocaine. Each OC was divided in 16 items recorded as visualized or not. Four scores were compared between both groups: out of 16 (1 side), out of 32 (both sides), out of 12 concerning only the narrowest and highest bilateral spaces, and out of 4 to divide these specific areas in anterior, middle, and posterior parts. Length of procedure, pain, epistaxis, blood mark on the endosheath, sneezing, rhinorrhea, and causes of failure were recorded. RESULTS: There was no significant difference between both groups concerning score out of 16 or 32. The visibility of the narrower and higher spaces was better in group 2: scores out of 12 were significantly different between the groups (P = .025), as were scores out of 4 for the anterior and middle parts of the OC (P = .02 and .01 respectively). Morbidity was low without differences between the groups. Deviated nasal septum was the only cause of failure and increased the patients' pain during the examination (P = .045). CONCLUSION: Nasal fibroscopy could be used to explore the different portions of the OC efficiently and with low morbidity. PMID- 21622803 TI - Response to: Weight gain after tonsillectomy. PMID- 21622805 TI - Multicenter study of a novel adjustable tongue-advancement device for obstructive sleep apnea. PMID- 21622807 TI - Racial/ethnic variations in the incidence and evolutionary pathogenesis of otitis media. PMID- 21622808 TI - Postmatch findings in otolaryngology residency applicants with publication misrepresentation. PMID- 21622811 TI - Differential molecular responses of rice and wheat coleoptiles to anoxia reveal novel metabolic adaptations in amino acid metabolism for tissue tolerance. AB - Rice (Oryza sativa) and wheat (Triticum aestivum) are the most important starch crops in world agriculture. While both germinate with an anatomically similar coleoptile, this tissue defines the early anoxia tolerance of rice and the anoxia intolerance of wheat seedlings. We combined protein and metabolite profiling analysis to compare the differences in response to anoxia between the rice and wheat coleoptiles. Rice coleoptiles responded to anoxia dramatically, not only at the level of protein synthesis but also at the level of altered metabolite pools, while the wheat response to anoxia was slight in comparison. We found significant increases in the abundance of proteins in rice coleoptiles related to protein translation and antioxidant defense and an accumulation of a set of enzymes involved in serine, glycine, and alanine biosynthesis from glyceraldehyde-3 phosphate or pyruvate, which correlates with an observed accumulation of these amino acids in anoxic rice. We show a positive effect on wheat root anoxia tolerance by exogenous addition of these amino acids, indicating that their synthesis could be linked to rice anoxia tolerance. The potential role of amino acid biosynthesis contributing to anoxia tolerance in cells is discussed. PMID- 21622812 TI - Virus-induced necrosis is a consequence of direct protein-protein interaction between a viral RNA-silencing suppressor and a host catalase. AB - Many plant host factors are known to interact with viral proteins during pathogenesis, but how a plant virus induces a specific disease symptom still needs further research. A lily strain of Cucumber mosaic virus (CMV-HL) can induce discrete necrotic spots on infected Arabidopsis (Arabidopsis thaliana) plants; other CMV strains can induce similar spots, but they are not as distinct as those induced by CMV-HL. The CMV 2b protein (2b), a known RNA-silencing suppressor, is involved in viral movement and symptom induction. Using in situ proximity ligation assay immunostaining and the protoplast assays, we report here that CMV 2b interacts directly with Catalase3 (CAT3) in infected tissues, a key enzyme in the breakdown of toxic hydrogen peroxide. Interestingly, CAT3, normally localized in the cytoplasm (glyoxysome), was recruited to the nucleus by an interaction between 2b and CAT3. Although overexpression of CAT3 in transgenic plants decreased the accumulation of CMV and delayed viral symptom development to some extent, 2b seems to neutralize the cellular catalase contributing to the host defense response, thus favoring viral infection. Our results thus provide evidence that, in addition to altering the type of symptom by disturbing microRNA pathways, 2b can directly bind to a host factor that is important in scavenging cellular hydrogen peroxide and thus interfere specifically with that host factor, leading to the induction of a specific necrosis. PMID- 21622813 TI - Drug-induced torsade de pointes arrhythmias in the chronic AV block dog are perpetuated by focal activity. AB - BACKGROUND: The electrically remodeled canine heart after chronic AV block (CAVB) has a high susceptibility for drug-induced torsade de pointes (TdP) arrhythmias. Although focal mechanisms have been considered for initiation, there is still controversy about whether reentry is the dominant mechanism for perpetuation of TdP. In this animal model with known nonuniform prolongation of repolarization, the mechanism of perpetuation of TdP arrhythmia was explored. METHODS AND RESULTS: Seventeen TdP-sensitive CAVB and 10 sinus rhythm (SR) dogs were studied. In 6 animals, 66 needle electrodes were evenly distributed transmurally to record 240 unipolar local electrograms simultaneously. Activation times and activation recovery intervals were determined before and during ibutilide-induced TdP. In 12 CAVB and 9 SR dogs, left ventricular (LV) and right ventricular (RV) epicardial electrograms were recorded with a 208-point multiterminal grid electrode allowing conduction velocity (CV) and ventricular effective refractory period (VERP) measurements. Biopsy specimens were processed for connexin43 (Cx43) expression and collagen content. Ventricular myocytes were isolated to determine sodium current (I(Na)) density and cell dimensions. Computer simulations were used to assess the effects of changes therein. In CAVB, VERP and ARI were increased, whereas CV was unaltered in LV. Transversal but not longitudinal CV was increased in RV. I(Na) was reduced by 37% in LV but unaltered in RV. LV and RV cell size were increased, but collagen and Cx43 content remained unchanged. Simulations showed increase in CV of RV as a consequence of increased cell size at normal I(Na). Ibutilide increased ARI, ERP, and maximal transmural dispersion of ERP (45 +/- 25 to 120 +/- 65 ms; P < 0.05). Twenty-eight of 47 episodes of self terminating TdP (43 +/- 72 beats) were analyzed. The majority (> 90%) of beats were focal; reentry was observed only occasionally. CONCLUSIONS: Focal activity is the dominant mechanism involved in perpetuation of ibutilide-induced TdP in CAVB dogs based on detailed 3D mapping. This conclusion is in line with unaltered conduction and documented increase in VERP. PMID- 21622814 TI - Dynamic ankle control in athletes with ankle instability during sports maneuvers. AB - BACKGROUND: Ankle sprain is a common sports injury. While the effects of static constraints in stabilizing the ankle joint are relatively well understood, those of dynamic constraints are less clear and require further investigation. PURPOSE: This study was undertaken to evaluate the dynamic stability of the ankle joint during the landing phase of running and stop-jump maneuvers in athletes with and without chronic ankle instability (CAI). STUDY DESIGN: Controlled laboratory study. METHODS: Fifteen athletes with CAI and 15 age-matched athletes without CAI performed running and stop-jump landing tasks. The dynamic ankle joint stiffness, tibialis anterior (TA)/peroneus longus (PL) and TA/gastrocnemius lateralis (GL) co-contraction indices, ankle joint angle, and root-mean-square (RMS) of the TA, PL, and GL electromyographic signals were measured during each task. RESULTS: During running, the CAI group exhibited a greater ankle inversion angle than the control group in the pre-landing phase (P = .012-.042) and a lower dynamic ankle joint stiffness in the post-landing phase (CAI: 0.109 +/- 0.039 N.m/deg; control: 0.150 +/- 0.068 N.m/deg; P = .048). In the stop-jump landing task, athletes with CAI had a significantly lower TA/PL co-contraction index during the pre-landing phase (CAI: 49.1 +/- 19; control: 64.8 +/- 16; P = .009). In addition, the CAI group exhibited a greater ankle inversion (P = .049), a lower peak eversion (P = .04), and a smaller RMS of the PL electromyographic signal in the post-landing phase (CAI: 0.73 +/- 0.32; control: 0.51 +/- 0.22; P = .04). CONCLUSION: Athletes with CAI had a relatively inverted ankle, reduced muscle co-contraction, and a lower dynamic stiffness in the ankle joint during the landing phase of sports maneuvers and this may jeopardize the stability of the ankle. CLINICAL RELEVANCE: Sports training or rehabilitation programs should differentiate between the pre landing and post-landing phases of sports maneuvers, and should educate athletes to land with an appropriate ankle position and muscle recruitment. PMID- 21622815 TI - Tibial fixation in anterior cruciate ligament reconstruction: a prospective randomized study comparing metal interference screw and staples with a centrally placed polyethylene screw and sheath. AB - BACKGROUND: The use of hamstring tendons for anterior cruciate ligament reconstruction has increased in popularity over recent years. However, concerns with the stability of graft fixation on the tibial side remain. Centrally placed interference screw/sheath implants have demonstrated promising results in biomechanical studies. HYPOTHESIS: Centrally placed, polyethylene screw and sheath implants will provide clinically equivalent fixation to the standard metal interference screw and supplemental staple fixation. STUDY DESIGN: Randomized controlled trial; Level of evidence, 1. METHODS: A total of 113 consecutive patients undergoing isolated, unilateral, primary anterior cruciate ligament reconstruction with hamstring autografts were randomized to tibial fixation with metal interference screw and staples (RCI) or with a centrally placed polyethylene screw and sheath implant (INTRAFIX). Prospective assessment of subjective outcomes was performed using Lysholm, Mohtadi, and International Knee Documentation Committee (IKDC) scores. RESULTS: At minimum 2-year follow-up, there were no significant differences between the 2 groups in terms of instrumented stability testing (KT-1000 arthrometer) or subjective assessment of knee outcomes (IKDC, Lysholm, Mohtadi). Both fixation methods demonstrated a significant, but not different, increase in outcomes scores from preoperative to postoperative evaluation. There were 7 failures (5 INTRAFIX, 2 RCI) caused by reinjury, but no statistically significant differences were observed between the 2 fixation methods. CONCLUSION: The centrally placed polyethylene screw and sheath provided equivalent clinical outcomes at minimum 2-year follow-up to standard tibial fixation with metal interference screw and staples. PMID- 21622816 TI - A new method to study changes in microvascular blood volume in muscle and adipose tissue: real-time imaging in humans and rat. AB - We employed and evaluated a new application of contrast-enhanced ultrasound for real-time imaging of changes in microvascular blood volume (MBV) in tissues in females, males, and rat. Continuous real-time imaging was performed using contrast-enhanced ultrasound to quantify infused gas-filled microbubbles in the microcirculation. It was necessary to infuse microbubbles for a minimum of 5-7 min to obtain steady-state bubble concentration, a prerequisite for making comparisons between different physiological states. Insulin clamped at a submaximal concentration (~75 MUU/ml) increased MBV by 27 and 39% in females and males, respectively, and by 30% in female subcutaneous adipose tissue. There was no difference in the ability of insulin to increase muscle MBV in females and males, and microvascular perfusion rate was not increased significantly by insulin. However, perfusion rate of the microvascular space was higher in females compared with males. In rats, insulin clamped at a maximal concentration increased muscle MBV by 60%. Large increases in microvascular volume and perfusion rate were detected during electrical stimulation of muscle in rats and immediately after exercise in humans. We have demonstrated that real-time imaging of changes in MBV is possible in human and rat muscle and in subcutaneous adipose tissue and that the method is sensitive enough to pick up relatively small changes in MBV when performed with due consideration of steady-state microbubble concentration. Because of real-time imaging, the method has wide applications for determining MBV in different organs during various physiological or pathophysiological conditions. PMID- 21622817 TI - Effects of carbenoxolone on flow-mediated vasodilatation in healthy adults. AB - Gap junctions play a key role in maintaining the functional integrity of the vascular wall. Using carbenoxolone (CBX) as a gap junction blocker, we aimed to assess the contribution of gap junctions in the vascular wall to flow-mediated vasodilatation (FMD) in healthy adults. Percentage FMD (%FMD) and circulating vasoactive molecules/activity, including atrial natriuretic peptide (ANP), B-type natriuretic peptide (BNP), aldosterone, cortisol, plasma renin activity (PRA), and endothelin (ET-1), were measured in 25 healthy volunteers (mean age: 30.1 +/- 5.4 yr; 14 males) before and after oral administration of CBX (100 mg). %FMD decreased after ingestion of CBX (9.71 +/- 3.1 vs. 3.40 +/- 2.0%; P < 0.0001). The levels of ANP, BNP, cortisol, and ET-1 remained stationary, while both PRA and aldosterone decreased (P < 0.005) after CBX ingestion. Blood pressure and heart rate were minimally changed by CBX. Inhibition of gap junctional communication by CBX impairs FMD in healthy persons, suggesting that physiologically, vascular gap junctions participate in the maintenance of FMD. CBX does not induce the release of vasoconstricting molecules or enhance vasoconstriction, suggesting that inhibition of gap junctional communication by CBX underlies the impairment of FMD. Therefore, administering CBX in FMD examination can be a way to follow the effect of gap junctions on endothelial function, but further work remains to verify the specificity of CBX effect. PMID- 21622818 TI - Modulating endothelial nitric oxide synthase: a new cardiovascular therapeutic strategy. AB - The pathogenesis of many cardiovascular diseases is associated with reduced nitric oxide (NO) bioavailability and/or increased endothelial NO synthase (eNOS) dependent superoxide formation. These findings support that restoring and conserving adequate NO signaling in the heart and blood vessels is a promising therapeutic intervention. In particular, modulating eNOS, e.g., through increasing the bioavailability of its substrate and cofactors, enhancing its transcription, and interfering with other modulators of eNOS pathway, such as netrin-1, has a high potential for effective treatments of cardiovascular diseases. This review provides an overview of the possibilities for modulating eNOS and how this may be translated to the clinic in addition to describing the genetic models used to study eNOS modulation. PMID- 21622819 TI - Both AT1 and AT2 receptors mediate proliferation and migration of porcine vascular smooth muscle cells. AB - Angiotensin receptor antagonists have shown clinical promise in modulating vascular disease, in part by limiting smooth muscle cell proliferation and migration. The majority of studies examining the contribution of these receptors have been undertaken in cells derived from rat aorta, which primarily express the ANG II type 1 (AT(1)) receptor. This investigation studied the relative contribution of AT(1) and ANG II type 2 (AT(2)) receptors to the mitogenic program of porcine smooth muscle cells. Smooth muscle cells were derived from porcine coronary artery explants. The presence of both AT(1) and AT(2) receptors was demonstrated through ligand binding and RT-PCR analysis. Biochemical and cellular markers of proliferation were monitored in the presence of selective receptor antagonists. Smooth muscle cell migration was measured using both wound healing and Boyden chamber migration assays. Visualization of the AT(1) and AT(2) receptors in growing and quiescent porcine smooth muscle cells with epifluorescence microscopy demonstrated that their subcellular distribution varied with growth state. An examination with several growth assays revealed that both AT(1)-specific losartan and AT(2)-specific PD-123319 receptor antagonists inhibited ANG II-stimulated RNA and DNA synthesis, PCNA expression, and hyperplasia. ANG II induced both directional and nondirectional cell migration. AT(1) receptor antagonist treatment significantly decreased ANG II-induced directional migration only, whereas AT(2) receptor antagonist treatment significantly reduced both modes of migration. Interestingly, the focal adhesion kinase inhibitor PF-573228 also blocked migration but not proliferation. Furthermore, focal adhesion kinase activation in response to ANG II was prevented only by PD-123319, indicating that this activation is downstream of the AT(2) receptor. The observed role of the AT(2) receptor in ANG II-induced migration was confirmed with smooth muscle cells depleted of the AT(2) receptor with short hairpin RNA treatment. PMID- 21622820 TI - Validation of a patient-specific one-dimensional model of the systemic arterial tree. AB - The aim of this study is to develop and validate a patient-specific distributed model of the systemic arterial tree. This model is built using geometric and hemodynamic data measured on a specific person and validated with noninvasive measurements of flow and pressure on the same person, providing thus a patient specific model and validation. The systemic arterial tree geometry was obtained from MR angiographic measurements. A nonlinear viscoelastic constitutive law for the arterial wall is considered. Arterial wall distensibility is based on literature data and adapted to match the wave propagation velocity of the main arteries of the specific subject, which were estimated by pressure waves traveling time. The intimal shear stress is modeled using the Witzig-Womersley theory. Blood pressure is measured using applanation tonometry and flow rate using transcranial ultrasound and phase-contrast-MRI. The model predicts pressure and flow waveforms in good qualitative and quantitative agreement with the in vivo measurements, in terms of wave shape and specific wave features. Comparison with a generic one-dimensional model shows that the patient-specific model better predicts pressure and flow at specific arterial sites. These results obtained let us conclude that a patient-specific one-dimensional model of the arterial tree is able to predict well pressure and flow waveforms in the main systemic circulation, whereas this is not always the case for a generic one-dimensional model. PMID- 21622821 TI - Biochemical and myofilament responses of the right ventricle to severe pulmonary hypertension. AB - Right ventricular (RV) failure is one of the strongest predictors of mortality both in the presence of left ventricular decompensation and in the context of pulmonary vascular disease. Despite this, there is a limited understanding of the biochemical and mechanical characteristics of the pressure-overloaded RV at the level of the cardiac myocyte. To better understand this, we studied ventricular muscle obtained from neonatal calves that were subjected to hypobaric atmospheric conditions, which result in profound pulmonary hypertension. We found that RV pressure overload resulted in significant changes in the phosphorylation of key contractile proteins. Total phosphorylation of troponin I was decreased with pressure overload, predominantly reflecting changes at the putative PKA site at Ser(22/23). Similarly, both troponin T and myosin light chain 2 showed a significant decline in phosphorylation. Desmin was unchanged, and myosin-binding protein C (MyBP-C) phosphorylation was apparently increased. However, the apparent increase in MyBP-C phosphorylation was not due to phosphorylation but rather to an increase in MyBP-C total protein. Importantly, these findings were seen in all regions of the RV and were paralleled by reduced Ca(2+) sensitivity with preserved maximal Ca(2+) saturated developed force normalized to cross sectional area in isolated skinned right ventricular myocyte fragments. No changes in total force or cooperativity were seen. Taken together, these results suggest that RV failure is mechanistically unique from left ventricular failure. PMID- 21622822 TI - Increasing dihydrobiopterin causes dysfunction of endothelial nitric oxide synthase in rats in vivo. AB - An elevation of oxidized forms of tetrahydrobiopterin (BH(4)), especially dihydrobiopterin (BH(2)), has been reported in the setting of oxidative stress, such as arteriosclerotic/atherosclerotic disorders, where endothelial nitric oxide synthase (eNOS) is dysfunctional, but the role of BH(2) in the regulation of eNOS activity in vivo remains to be evaluated. This study was designed to clarify whether increasing BH(2) concentration causes endothelial dysfunction in rats. To increase vascular BH(2) levels, the BH(2) precursor sepiapterin (SEP) was intravenously given after the administration of the specific dihydrofolate reductase inhibitor methotrexate (MTX) to block intracellular conversion of BH(2) to BH(4). MTX/SEP treatment did not significantly affect aortic BH(4) levels compared with control treatment. However, MTX/SEP treatment markedly augmented aortic BH(2) levels (291.1 +/- 29.2 vs. 33.4 +/- 6.4 pmol/g, P < 0.01) in association with moderate hypertension. Treatment with MTX alone did not significantly alter blood pressure or BH(4) levels but decreased the BH(4)-to BH(2) ratio. Treatment with MTX/SEP, but not with MTX alone, impaired ACh-induced vasodilator and depressor responses compared with the control treatment (both P < 0.05) and also aggravated ACh-induced endothelium-dependent relaxations (P < 0.05) of isolated aortas without affecting sodium nitroprusside-induced endothelium-independent relaxations. Importantly, MTX/SEP treatment significantly enhanced aortic superoxide production, which was diminished by NOS inhibitor treatment, and the impaired ACh-induced relaxations were reversed with SOD (P < 0.05), suggesting the involvement of eNOS uncoupling. These results indicate, for the first time, that increasing BH(2) causes eNOS dysfunction in vivo even in the absence of BH(4) deficiency, demonstrating a novel insight into the regulation of endothelial function. PMID- 21622823 TI - Impact of elevated uric acid on ventricular remodeling in infarcted rats with experimental hyperuricemia. AB - Hyperuricemia is associated with cardiovascular disease, but it is usually considered a marker rather than a risk factor. Previous studies using uric acid lowering drugs in normouricemic animals are not suitable to answer the effect of hyperuricemia on ventricular remodeling after myocardial infarction. The purpose of this study was to determine whether hyperuricemia adversely affects ventricular remodeling in infarcted rats with elevated uric acid. Male Wistar rats aged 8 wk were randomly assigned into either vehicle, oxonic acid, oxonic acid + allopurinol, oxonic acid + benzbromarone, oxonic acid + ABT-627, or oxonic acid + tempol for 4 wk starting 24 h after ligation. Postinfarction was associated with increased oxidant production, as measured by myocardial superoxide, isoprostane, xanthine oxidase activity, and dihydroethidium staining. Compared with normouricemic infarcted rats, hyperuricemic infarcted rats had a significant increase of superoxide production (1.7*) and endothelin-1 protein (1.2*) and mRNA (1.4*) expression, which was associated with increased left ventricular dysfunction and enhanced myocardial hypertrophy and fibrosis. These changes were all prevented by treatment with allopurinol. For similar levels of urate lowering, the uricosuric agent benzbromarone had no effect on ventricular remodeling. In spite of equivalent hyperuricemia, the ability of both ABT-627 and tempol to attenuate ventricular remodeling suggested involvement of endothelin-1 and redox pathways. Hyperuricemia is associated with unfavorable ventricular remodeling probably through a superoxide and endothelin-1-dependent pathway. Uric acid lowering without inhibition of superoxide and endothelin-1 may not have an effect on remodeling. Chronic administration of allopurinol, ABT-627, and tempol is associated with attenuated ventricular remodeling. PMID- 21622824 TI - Aerobic exercise reverses arterial inflammation with aging in mice. AB - We tested the hypothesis that regular aerobic exercise reverses arterial inflammation with aging. When compared with young controls (6.2 +/- 0.4 mo; n = 7), old (31.3 +/- 0.5 mo; n = 11) male B6D2F1 cage-restricted mice demonstrated increased arterial activation of the proinflammatory transcription factor NF kappaB, as indicated by greater aortic phosphorylation of both the inhibitor of NF-kappaB kinase (IKK) and the p65 subunit of NF-kappaB (both P < 0.05). Similarly, aortic expression of the proinflammatory cytokines IL-1 and IL-6, IFN gamma, and TNF-alpha were greater in the old mice (all P < 0.05). Macrophage and T lymphocyte abundance was unchanged with age in the aortic intima and media but was markedly increased in the adventitia and perivascular fat tissue of old mice (all P < 0.05). This proinflammatory arterial phenotype with aging was associated with vascular dysfunction, as reflected by impaired nitric oxide-mediated endothelium-dependent dilation. Voluntary wheel running (10-14 wk) normalized aortic IKK-NF-kappaB activation, cytokine expression, adventitial and perivascular macrophage infiltration, and vascular function in old mice (32.4 +/- 0.3 mo; n = 8) while having no consistent effects in young mice. Short-term voluntary wheel running started late in life reverses arterial inflammation with aging in mice possibly via outside-in actions. These anti-inflammatory effects may play an important role in the amelioration of age-associated vascular dysfunction by regular aerobic exercise. PMID- 21622825 TI - Ascorbate improves circulation in postural tachycardia syndrome. AB - Low flow postural tachycardia syndrome (LFP) is associated with vasoconstriction, reduced cardiac output, increased plasma angiotensin II, reduced bioavailable nitric oxide (NO), and oxidative stress. We tested whether ascorbate would improve cutaneous NO and reduce vasoconstriction when delivered systemically. We used local cutaneous heating to 42 degrees C and laser Doppler flowmetry to assess NO-dependent conductance (%CVC(max)) to sodium ascorbate and the systemic hemodynamic response to ascorbic acid in 11 LFP patients and in 8 control subjects (aged 23 +/- 2 yr). We perfused intradermal microdialysis catheters with sodium ascorbate (10 mM) or Ringer solution. Predrug heat response was reduced in LFP, particularly the NO-dependent plateau phase (56 +/- 6 vs. 88 +/- 7%CVC(max)). Ascorbate increased baseline skin flow in LFP and control subjects and increased the LFP plateau response (82 +/- 6 vs. 92 +/- 6 control). Systemic infusion experiments used Finometer and ModelFlow to estimate relative cardiac index (CI) and forearm and calf venous occlusion plethysmography to estimate blood flows, peripheral arterial and venous resistances, and capacitance before and after infusing ascorbic acid. CI increased 40% after ascorbate as did peripheral flows. Peripheral resistances were increased (nearly double control) and decreased by nearly 50% after ascorbate. Calf capacitance and venous resistance were decreased compared with control but normalized with ascorbate. These data provide experimental support for the concept that oxidative stress and reduced NO possibly contribute to vasoconstriction and venoconstriction of LFP. PMID- 21622826 TI - Assessment of coronary microcirculation in a swine animal model. AB - Coronary microvascular dysfunction has important prognostic implications. Several hemodynamic indexes, such as coronary flow reserve (CFR), microvascular resistance, and zero-flow pressure (P(zf)), were used to establish the most reliable index to assess coronary microcirculation. Fifteen swine were instrumented with a flow probe, and a pressure wire was advanced into the distal left anterior descending artery. Adenosine was used to produce maximum hyperemia. Microspheres were used to create microvascular dysfunction. An occluder was used to produce stenosis. Blood flow from the probe (Q(p)), aortic pressure, distal coronary pressure, and right atrium pressure were recorded. Angiographic flow (Q(a)) was calculated using a time-density curve. Flow probe-based CFR and angiographic CFR were calculated using Q(p) and Q(a), respectively. Flow probe based (NMR(qh)) and angiographic normalized microvascular resistance (NMR(ah)) were determined using Q(p) and Q(a), respectively, during hyperemia. P(zf) was calculated using Q(p) and distal coronary pressure. Two series of receiver operating characteristic curves were generated: normal epicardial artery model (N model) and stenosis model (S model). The areas under the receiver operating characteristic curves for flow probe-based CFR, angiographic CFR, NMR(qh), NMR(ah), and P(zf) were 0.855, 0.836, 0.976, 0.956, and 0.855 in N model and 0.737, 0.700, 0.935, 0.889, and 0.698 in S model. Both NMR(qh) and NMR(ah) were significantly more reliable than CFR and P(zf) in detecting the microvascular deterioration. Compared with CFR and P(zf), NMR provided a more accurate assessment of microcirculation. This improved accuracy was more prevalent when stenosis existed. Moreover, NMR(ah) is potentially a less invasive method for assessing coronary microcirculation. PMID- 21622827 TI - 2',3'-cAMP, 3'-AMP, and 2'-AMP inhibit human aortic and coronary vascular smooth muscle cell proliferation via A2B receptors. AB - Rat vascular smooth muscle cells (VSMCs) from renal microvessels metabolize 2',3' cAMP to 2'-AMP and 3'-AMP, and these AMPs are converted to adenosine that inhibits microvascular VSMC proliferation via A(2B) receptors. The goal of this study was to test whether this mechanism also exists in VSMCs from conduit arteries and whether it is similarly expressed in human vs. rat VSMCs. Incubation of rat and human aortic VSMCs with 2',3'-cAMP concentration-dependently increased levels of 2'-AMP and 3'-AMP in the medium, with a similar absolute increase in 2' AMP vs. 3'-AMP. In contrast, in human coronary VSMCs, 2',3'-cAMP increased 2'-AMP levels yet had little effect on 3'-AMP levels. In all cell types, 2',3'-cAMP increased levels of adenosine, but not 5'-AMP, and 2',3'-AMP inhibited cell proliferation. Antagonism of A(2B) receptors (MRS-1754), but not A(1) (1,3 dipropyl-8-cyclopentylxanthine), A(2A) (SCH-58261), or A(3) (VUF-5574) receptors, attenuated the antiproliferative effects of 2',3'-cAMP. In all cell types, 2' AMP, 3'-AMP, and 5'-AMP increased adenosine levels, and inhibition of ecto-5' nucleotidase blocked this effect of 5'-AMP but not that of 2'-AMP nor 3'-AMP. Also, 2'-AMP, 3'-AMP, and 5'-AMP, like 2',3'-cAMP, exerted antiproliferative effects that were abolished by antagonism of A(2B) receptors with MRS-1754. In conclusion, VSMCs from conduit arteries metabolize 2',3'-cAMP to AMPs, which are metabolized to adenosine. In rat and human aortic VSMCs, both 2'-AMP and 3'-AMP are involved in this process, whereas, in human coronary VSMCs, 2',3'-cAMP is mainly converted to 2'-AMP. Because adenosine inhibits VSMC proliferation via A(2B) receptors, local vascular production of 2',3'-cAMP may protect conduit arteries from atherosclerosis. PMID- 21622828 TI - Repeated sauna therapy attenuates ventricular remodeling after myocardial infarction in rats by increasing coronary vascularity of noninfarcted myocardium. AB - Repeated sauna therapy (ST) increases endothelial nitric oxide synthase (eNOS) activity and improves cardiac function in heart failure as well as peripheral blood flow in ischemic limbs. The present study investigates whether ST can increase coronary vascularity and thus attenuate cardiac remodeling after myocardial infarction (MI). We induced MI by ligating the left coronary artery of Wistar rats. The rats were placed in a far-infrared dry sauna at 41 degrees C for 15 min and then at 34 degrees C for 20 min once daily for 4 wk. Cardiac hemodynamic, histopathological, and gene analyses were performed. Despite the similar sizes of MI between the ST and non-ST groups (51.4 +/- 0.3 vs. 51.1 +/- 0.2%), ST reduced left ventricular (LV) end-diastolic (9.7 +/- 0.4 vs. 10.7 +/- 0.5 mm, P < 0.01) and end-systolic (8.6 +/- 0.5 vs. 9.6 +/- 0.6 mm, P < 0.01) dimensions and attenuated MI-induced increases in LV end-diastolic pressure. Cross-sectional areas of cardiomyocytes were smaller in ST rats and associated with a significant reduction in myocardial atrial natriuretic peptide mRNA levels. Vascular density was reduced in the noninfarcted myocardium of non-ST rats, and the density of cells positive for CD31 and for alpha-smooth muscle actin was decreased. These decreases were attenuated in ST rats compared with non ST rats and associated with increases in myocardial eNOS and vascular endothelial growth factor mRNA levels. In conclusion, ST attenuates cardiac remodeling after MI, at least in part, through improving coronary vascularity in the noninfarcted myocardium. Repeated ST might serve as a novel noninvasive therapy for patients with MI. PMID- 21622829 TI - Histamine receptor H1 in the nucleus tractus solitarii regulates arterial pressure and heart rate in rats. AB - Axons of histamine (HA)-containing neurons are known to project from the posterior hypothalamus to many areas of the brain, including the nucleus tractus solitarii (NTS), a central brain structure that plays an important role in regulating arterial pressure. However, the functional significance of NTS HA is still not fully established. In this study, we microinjected HA or 2 pyridylethylamine, a HA-receptor H(1)-specific agonist, into the NTS of urethane anesthetized Wister rats to identify the potential functions of NTS HA on cardiovascular regulation. When HA or H(1)-receptor-specific agonist was bilaterally microinjected into the NTS, mean arterial pressure (MAP) and heart rate (HR) were significantly increased, whereas pretreatment with the H(1) receptor-specific antagonist cetirizine into the NTS significantly inhibited the cardiovascular responses. The maximal responses of MAP and HR changes induced by HA or H(1)-receptor-specific agonist were dose dependent. We also confirmed gene expression of HA receptors in the NTS and that the expression level of H(1) mRNA was higher than that of the other subtypes. In addition, we found that H(1) receptors are mainly expressed in neurons of the NTS. These findings suggested that HA within the NTS may play a role in regulating cardiovascular homeostasis via activation of H(1) receptors expressed in the NTS neurons. PMID- 21622830 TI - Impact of exercise training on endothelial transcriptional profiles in healthy swine: a genome-wide microarray analysis. AB - While the salutary effects of exercise training on conduit artery endothelial cells have been reported in animals and humans with cardiovascular risk factors or disease, whether a healthy endothelium is alterable with exercise training is less certain. The purpose of this study was to evaluate the impact of exercise training on transcriptional profiles in normal endothelial cells using a genome wide microarray analysis. Brachial and internal mammary endothelial gene expression was compared between a group of healthy pigs that exercise trained for 16-20 wk (n = 8) and a group that remained sedentary (n = 8). We found that a total of 130 genes were upregulated and 84 genes downregulated in brachial artery endothelial cells with exercise training (>1.5-fold and false discovery rate <15%). In contrast, a total of 113 genes were upregulated and 31 genes downregulated in internal mammary artery endothelial cells using the same criteria. Although there was an overlap of 66 genes (59 upregulated and 7 downregulated with exercise training) between the brachial and internal mammary arteries, the identified endothelial gene networks and biological processes influenced by exercise training were distinctly different between the brachial and internal mammary arteries. These data indicate that a healthy endothelium is indeed responsive to exercise training and support the concept that the influence of physical activity on endothelial gene expression is not homogenously distributed throughout the vasculature. PMID- 21622831 TI - Rho-kinase: important new therapeutic target in cardiovascular diseases. AB - Rho-kinase (ROCKs) belongs to the family of serine/threonine kinases and is an important downstream effector of the small GTP-binding protein RhoA. There are two isoforms of Rho-kinase, ROCK1 and ROCK2, and they have different functions with ROCK1 for circulating inflammatory cells and ROCK2 for vascular smooth muscle cells. It has been demonstrated that the RhoA/Rho-kinase pathway plays an important role in various fundamental cellular functions, including contraction, motility, proliferation, and apoptosis, leading to the development of cardiovascular disease. The important role of Rho-kinase in vivo has been demonstrated in the pathogenesis of vasospasm, arteriosclerosis, ischemia reperfusion injury, hypertension, pulmonary hypertension, stroke, and heart failure. Furthermore, the beneficial effects of fasudil, a selective Rho-kinase inhibitor, have been demonstrated for the treatment of several cardiovascular diseases in humans. Thus the Rho-kinase pathway is an important new therapeutic target in cardiovascular medicine. PMID- 21622832 TI - Cardiac remodeling caused by transgenic overexpression of a corn Rac gene. AB - Rac1-GTPase activation plays a key role in the development and progression of cardiac remodeling. Therefore, we engineered a transgenic mouse model by overexpressing cDNA of a constitutively active form of Zea maize Rac gene (ZmRacD) specifically in the hearts of FVB/N mice. Echocardiography and MRI analyses showed cardiac hypertrophy in old transgenic mice, as evidenced by increased left ventricular (LV) mass and LV mass-to-body weight ratio, which are associated with relative ventricular chamber dilation and systolic dysfunction. LV hypertrophy in the hearts of old transgenic mice was further confirmed by an increased heart weight-to-body weight ratio and histopathology analysis. The cardiac remodeling in old transgenic mice was coupled with increased myocardial Rac-GTPase activity (372%) and ROS production (462%). There were also increases in alpha(1)-integrin (224%) and beta(1)-integrin (240%) expression. This led to the activation of hypertrophic signaling pathways, e.g., ERK1/2 (295%) and JNK (223%). Pravastatin treatment led to inhibition of Rac-GTPase activity and integrin signaling. Interestingly, activation of ZmRacD expression with thyroxin led to cardiac dilation and systolic dysfunction in adult transgenic mice within 2 wk. In conclusion, this is the first study to show the conservation of Rho/Rac proteins between plant and animal kingdoms in vivo. Additionally, ZmRacD is a novel transgenic model that gradually develops a cardiac phenotype with aging. Furthermore, the shift from cardiac hypertrophy to dilated hearts via thyroxin treatment will provide us with an excellent system to study the temporal changes in cardiac signaling from adaptive to maladaptive hypertrophy and heart failure. PMID- 21622833 TI - Aquimarina agarilytica sp. nov., an agarolytic species isolated from a red alga. AB - A novel yellow-pigmented, agarolytic bacterial strain, designated ZC1T, was isolated from the surface of the marine red alga Porphyra haitanensis collected near Nan Ao Island, Guangdong province, China. The isolate was Gram-stain negative, strictly aerobic and rod-shaped and displayed beta-galactosidase, alkaline phosphatase, catalase and oxidase activities. The predominant cellular fatty acids were iso-C15:0, summed feature 3 (comprising C16:1omega7c and/or iso C15:0 2-OH) and iso-C17:0 3-OH. The major menaquinone was menaquinone 6 (MK-6). The DNA G+C content was 32.8 mol%. Phylogenetic analysis of the 16S rRNA gene sequence revealed that strain ZC1T was closely related to members of the genus Aquimarina in the family Flavobacteriaceae, phylum Bacteroidetes. Based on phylogenetic and phenotypic evidence, strain ZC1T (=CCTCC AB 2010229T=NBRC 107695T) represents the type strain of a novel species in the genus Aquimarina, for which the name Aquimarina agarilytica sp. nov. is proposed. PMID- 21622834 TI - Streptomyces nanhaiensis sp. nov., a marine streptomycete isolated from a deep sea sediment. AB - A novel aerobic streptomycete, strain SCSIO 01248T, was isolated from a sample of deep-sea sediment collected from the northern South China Sea, at a depth of 1632 m. This isolate formed yellow-white substrate mycelium and grey-white aerial hyphae. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain SCSIO 01248T was most closely related to Streptomyces radiopugnans R97T (98.8 % sequence similarity), S. macrosporus NBRC 14748T (97.5 %) and S. megasporus NBRC 14749T (97.3 %). The novel strain could, however, be readily differentiated from S. radiopugnans DSM 41901T on the basis of some physiological and cellular chemical characteristics; the level of DNA-DNA relatedness between these two strains was only 40 %. Based on phylogenetic and phenotypic evidence, strain SCSIO 01248T represents a novel species, for which the name Streptomyces nanhaiensis sp. nov. is proposed. The type strain is SCSIO 01248T (=DSM 41926T=KCTC 19401T=CCTCC AA 208007T). PMID- 21622835 TI - Lactobacillus xiangfangensis sp. nov., isolated from Chinese pickle. AB - A Gram-positive bacterial strain, 3.1.1T, was isolated from traditional pickle in Heilongjiang Province, China. The bacterium was characterized by a polyphasic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, dnaK gene sequence analysis, fatty acid methyl ester analysis, determination of DNA G+C content, DNA-DNA hybridization and an analysis of phenotypic features. Based upon the data obtained in the present study, a novel species, Lactobacillus xiangfangensis sp. nov., is proposed with the type strain 3.1.1T (=LMG 26013T=NCIMB 14687T). PMID- 21622836 TI - Streptomyces gramineus sp. nov., an antibiotic-producing actinobacterium isolated from bamboo (Sasa borealis) rhizosphere soil. AB - Two actinobacterial strains, JR-43T and JR-4, were isolated from bamboo (Sasa borealis) rhizosphere soil. The isolates produced grey aerial mycelium and a yellow soluble pigment on ISP 4. Microscopic observation revealed that strains JR 43T and JR-4 produced rectiflexibiles spore chains with spiny surfaces. Both isolates had antibacterial activity against plant-pathogenic bacteria, such as Xanthomonas campestris LMG 568T and Xanthomonas axonopodis pv. vesicatoria LMG 905. The isolates contained iso-C14:0, iso-C15:0, anteiso-C15:0 and iso-C16:0 as the major fatty acids and MK-9(H6) and MK-9(H8) as the major isoprenoid quinones. Phylogenetic analysis of the 16S rRNA gene sequences of strains JR-43T and JR-4 showed that they grouped within Streptomyces cluster II and had highest sequence similarity to Streptomyces seoulensis NBRC 16668T and Streptomyces recifensis NBRC 12813T (both 98.2 % 16S rRNA gene sequence similarity). DNA-DNA relatedness between strain JR-43T and S. seoulensis NBRC 16668T and S. recifensis NBRC 12813T ranged from 31.42 to 42.92 %. Based on DNA-DNA relatedness and morphological and phenotypic data, strains JR-43T and JR-4 could be distinguished from the type strains of phylogenetically related species. They are therefore considered to represent a novel species of the genus Streptomyces, for which the name Streptomyces gramineus sp. nov. is proposed. The type strain is JR-43T (=KACC 15079T=NBRC 107863T). Strain JR-4 (=KACC 15078= NBRC 107864) is a reference strain [corrected]. PMID- 21622837 TI - Brevibacillus aydinogluensis sp. nov., a moderately thermophilic bacterium isolated from Karakoc hot spring. AB - Two Gram-positive, moderately thermophilic, endospore-forming, rod-shaped, motile bacteria, designated PDF25T and PDF30, were isolated from Karakoc hot spring in the province of Izmir, Turkey, and were characterized in order to determine their phylogenetic positions. 16S rRNA gene sequence analysis revealed that the two strains belonged to the genus Brevibacillus; strain PDF25T showed highest sequence similarity to strain PDF30 (99.4 %) and Brevibacillus thermoruber DSM 7064T (98.5 %). The major fatty acids of strain PDF25T were iso-C15:0 (39.30 %), anteiso-C15:0 (26.10 %) and iso-C16:0 (14.75 %). Polar lipid analysis revealed diphosphatidylglycerol, phosphatidylglycerol, phosphatidylmonomethylethanolamine, phosphatidylethanolamine, and a variety of unidentified aminophospholipids, phospholipids and aminolipids. The major isoprenoid quinone was MK-7. The G+C content of the genomic DNA was 56.09 mol%. DNA-DNA hybridization experiments revealed 58 % relatedness between strain PDF25T and B. thermoruber DSM 7064T. Based on these data, the two strains are considered to represent a novel species of the genus Brevibacillus, for which the name Brevibacillus aydinogluensis sp. nov. is proposed. The type strain is PDF25T (=DSM 24395T=LMG 26289T). PMID- 21622838 TI - Fire alters patterns of genetic diversity among 3 lizard species in Florida Scrub habitat. AB - The Florida Sand Skink (Plestiodon reynoldsi), the Florida Scrub Lizard (Sceloporus woodi), and the Six-lined Racerunner (Aspidoscelis sexlineata) occur in the threatened and fire-maintained Florida scrub habitat. Fire may have different consequences to local genetic diversity of these species because they each have different microhabitat preference. We collected tissue samples of each species from 3 sites with different time-since-fire: Florida Sand Skink n = 73, Florida Scrub Lizard n = 70, and Six-lined Racerunner n = 66. We compared the effect of fire on genetic diversity at microsatellite loci for each species. We screened 8 loci for the Florida Sand Skink, 6 loci for the Florida Scrub Lizard, and 6 loci for the Six-lined Racerunner. We also tested 2 potential driving mechanisms for the observed change in genetic diversity, a metapopulation source/sink model and a local demographic model. Genetic diversity varied with fire history, and significant genetic differentiation occurred among sites. The Florida Scrub Lizard had highest genetic variation at more recently burned sites, whereas the Florida Sand Skink and the Six-lined Racerunner had highest genetic variation at less recently burned sites. Habitat preferences of the Florida Sand Skink and the Florida Scrub Lizard may explain their discordant results, and the Six-lined Racerunner may have a more complicated genetic response to fire or is acted on at a different geographic scale than we have investigated. Our results indicate that these species may respond to fire in a more complicated manner than predicted by our metapopulation model or local demographic model. Our results show that the population-level responses in genetic diversity to fire are species specific mandating conservation management of habitat diversity through a mosaic of burn frequencies. PMID- 21622839 TI - S-Nitrosoglutathione reductase (GSNOR) mediates the biosynthesis of jasmonic acid and ethylene induced by feeding of the insect herbivore Manduca sexta and is important for jasmonate-elicited responses in Nicotiana attenuata. AB - S-nitrosoglutathione reductase (GSNOR) reduces the nitric oxide (NO) adduct S nitrosoglutathione (GSNO), an essential reservoir for NO bioactivity. In plants, GSNOR has been found to be important in resistance to bacterial and fungal pathogens, but whether it is also involved in plant-herbivore interactions was not known. Using a virus-induced gene silencing (VIGS) system, the activity of GSNOR in a wild tobacco species, Nicotiana attenuata, was knocked down and the function of GSNOR in defence against the insect herbivore Manduca sexta was examined. Silencing GSNOR decreased the herbivory-induced accumulation of jasmonic acid (JA) and ethylene, two important phytohormones regulating plant defence levels, without compromising the activity of two mitogen-activated protein kinases (MAPKs), salicylic acid-induced protein kinase (SIPK) and wound induced protein kinase (WIPK). Decreased activity of trypsin proteinase inhibitors (TPIs) were detected in GSNOR-silenced plants after simulated M. sexta feeding and bioassays indicated that GSNOR-silenced plants have elevated susceptibility to M. sexta attack. Furthermore, GSNOR is required for methyl jasmonate (MeJA)-induced accumulation of defence-related secondary metabolites (TPI, caffeoylputrescine, and diterpene glycosides) but is not needed for the transcriptional regulation of JAZ3 (jasmonate ZIM-domain 3) and TD (threonine deaminase), indicating that GSNOR mediates certain but not all jasmonate inducible responses. This work highlights the important role of GSNOR in plant resistance to herbivory and jasmonate signalling and suggests the potential involvement of NO in plant-herbivore interactions. Our data also suggest that GSNOR could be a target of genetic modification for improving crop resistance to herbivores. PMID- 21622841 TI - A knotless method of securing the subcuticular suture. PMID- 21622842 TI - Sepiapterin improves angiogenesis of pulmonary artery endothelial cells with in utero pulmonary hypertension by recoupling endothelial nitric oxide synthase. AB - Persistent pulmonary hypertension of the newborn (PPHN) is associated with decreased blood vessel density that contributes to increased pulmonary vascular resistance. Previous studies showed that uncoupled endothelial nitric oxide (NO) synthase (eNOS) activity and increased NADPH oxidase activity resulted in marked decreases in NO bioavailability and impaired angiogenesis in PPHN. In the present study, we hypothesize that loss of tetrahydrobiopterin (BH4), a critical cofactor for eNOS, induces uncoupled eNOS activity and impairs angiogenesis in PPHN. Pulmonary artery endothelial cells (PAEC) isolated from fetal lambs with PPHN (HTFL-PAEC) or control lambs (NFL-PAEC) were used to investigate the cellular mechanisms impairing angiogenesis in PPHN. Cellular mechanisms were examined with respect to BH4 levels, GTP-cyclohydrolase-1 (GCH-1) expression, eNOS dimer formation, and eNOS-heat shock protein 90 (hsp90) interactions under basal conditions and after sepiapterin (Sep) supplementation. Cellular levels of BH4, GCH-1 expression, and eNOS dimer formation were decreased in HTFL-PAEC compared with NFL-PAEC. Sep supplementation decreased apoptosis and increased in vitro angiogenesis in HTFL-PAEC and ex vivo pulmonary artery sprouting angiogenesis. Sep also increased cellular BH4 content, NO production, eNOS dimer formation, and eNOS-hsp90 association and decreased the superoxide formation in HTFL-PAEC. These data demonstrate that Sep improves NO production and angiogenic potential of HTFL PAEC by recoupling eNOS activity. Increasing BH4 levels via Sep supplementation may be an important therapy for improving eNOS function and restoring angiogenesis in PPHN. PMID- 21622843 TI - Trapping of BMP receptors in distinct membrane domains inhibits their function in pulmonary arterial hypertension. AB - Bone morphogenetic proteins (BMPs) are pleiotrophic growth factors that influence diverse processes such as skeletal development, hematopoiesis, and neurogenesis. They play crucial roles in diseases such as pulmonary arterial hypertension (PAH). In PAH, mutants of the BMP type II receptors (BMPR2) were detected, and their functions were impaired during BMP signaling. It is thought that expression levels of these receptors determine the fate of BMP signaling, with low levels of expression leading to decreased Smad activation in PAH. However, our studies demonstrate, for the first time, that the localization of receptors on the plasma membrane, in this case BMPR2, was misdirected. Three BMPR2 mutants, D485G, N519K, and R899X, which are known to be involved in PAH, were chosen as our model system. Our results show that all three BMPR2 mutants decreased BMP-dependent Smad phosphorylation and Smad signaling. Although the three mutants reached the cell membrane and their expression was lower than that of BMPR2, they formed smaller clusters and associated differently with membrane domains, such as caveolae and clathrin-coated pits. The disruption of these domains restored the Smad signaling of D485G and N519K to the level of wild-type BMPR2, showing that these mutants were trapped in the domains, rather than just expressed at a lower level on the surface. Therefore, new treatment options for PAH should also target receptor localization, rather than just expression level. PMID- 21622844 TI - Mucous solids and liquid secretion by airways: studies with normal pig, cystic fibrosis human, and non-cystic fibrosis human bronchi. AB - To better understand how airways produce thick airway mucus, nonvolatile solids were measured in liquid secreted by bronchi from normal pig, cystic fibrosis (CF) human, and non-CF human lungs. Bronchi were exposed to various secretagogues and anion secretion inhibitors to induce a range of liquid volume secretion rates. In all three groups, the relationship of solids concentration (percent nonvolatile solids) to liquid volume secretion rate was curvilinear, with higher solids concentration associated with lower rates of liquid volume secretion. In contrast, the secretion rates of solids mass and water mass as functions of liquid volume secretion rates exhibited positive linear correlations. The y intercepts of the solids mass-liquid volume secretion relationships for all three groups were positive, thus accounting for the higher solids concentrations in airway liquid at low rates of secretion. Predictive models derived from the solids mass and water mass linear equations fit the experimental percent solids data for the three groups. The ratio of solids mass secretion to liquid volume secretion was 5.2 and 2.4 times higher for CF bronchi than for pig and non-CF bronchi, respectively. These results indicate that normal pig, non-CF human, and CF human bronchi produce a high-percent-solids mucus (>8%) at low rates of liquid volume secretion (<=1.0 MUl.cm(-2).h(-1)). However, CF bronchi produce mucus with twice the percent solids (~8%) of pig or non-CF human bronchi at liquid volume secretion rates >=4.0 MUl.cm(-2).h(-1). PMID- 21622846 TI - Bone marrow cells repair cigarette smoke-induced emphysema in rats. AB - The therapeutic potential of stem cells in chronic obstructive pulmonary disease is not well known although stem cell therapy is effective in models of other pulmonary diseases. We tested the capacities of bone marrow cells (BMCs), mesenchymal stem cells (MSCs), and conditioned media of MSCs (MSC-CM) to repair cigarette smoke-induced emphysema. Inbred female Lewis rats were exposed to cigarette smoke for 6 mo and then received BMCs, MSCs, or MSC-CM from male Lewis rats. For 2 mo after injection, the BMC treatment gradually alleviated the cigarette smoke-induced emphysema and restored the increased mean linear intercept. The BMC treatment significantly increased cell proliferation and the number of small pulmonary vessels, reduced apoptotic cell death, attenuated the mean pulmonary arterial pressure, and inhibited muscularization in small pulmonary vessels. However, only a few male donor cells were detected from 1 day to 1 mo after BMC administration. The MSCs and cell-free MSC-CM also induced the repair of emphysema and increased the number of small pulmonary vessels. Our data show that BMC, MSCs, and MSC-CM treatment repaired cigarette smoke-induced emphysema. The repair activity of these treatments is consistent with a paracrine effect rather than stem cell engraftment because most of the donor cells disappeared and because cell-free MSC-CM also induced the repair. PMID- 21622845 TI - Adenosine activation of A(2B) receptor(s) is essential for stimulated epithelial ciliary motility and clearance. AB - Mucociliary clearance, vital to lung clearance, is dependent on cilia beat frequency (CBF), coordination of cilia, and the maintenance of periciliary fluid. Adenosine, the metabolic breakdown product of ATP, is an important modulator of ciliary motility. However, the contributions of specific adenosine receptors to key airway ciliary motility processes are unclear. We hypothesized that adenosine modulates ciliary motility via activation of its cell surface receptors (A(1), A(2A), A(2B), or A(3)). To test this hypothesis, mouse tracheal rings (MTRs) excised from wild-type and adenosine receptor knockout mice (A(1), A(2A), A(2B), or A(3), respectively), and bovine ciliated bronchial epithelial cells (BBECs) were stimulated with known cilia activators, isoproterenol (ISO; 10 MUM) and/or procaterol (10 MUM), in the presence or absence of 5'-(N-ethylcarboxamido) adenosine (NECA), a nonselective adenosine receptor agonist [100 nM (A(1), A(2A), A(3)); 10 MUM (A(2B))], and CBF was measured. Cells and MTRs were also stimulated with NECA (100 nM or 10 MUM) in the presence and absence of adenosine deaminase inhibitor, erythro-9- (2-hydroxy-3-nonyl) adenine hydrochloride (10 MUM). Both ISO and procaterol stimulated CBF in untreated cells and/or MTRs from both wild type and adenosine knockout mice by ~3 Hz. Likewise, CBF significantly increased ~2-3 Hz in BBECs and wild-type MTRs stimulated with NECA. MTRs from A(1), A(2A), and A(3) knockout mice stimulated with NECA also demonstrated an increase in CBF. However, NECA failed to stimulate CBF in MTRs from A(2B) knockout mice. To confirm the mechanism by which adenosine modulates CBF, protein kinase activity assays were conducted. The data revealed that NECA-stimulated CBF is mediated by the activation of cAMP-dependent PKA. Collectively, these data indicate that purinergic stimulation of CBF requires A(2B) adenosine receptor activation, likely via a PKA-dependent pathway. PMID- 21622847 TI - Diaphragm muscle fiber function and structure in humans with hemidiaphragm paralysis. AB - Recent studies proposed that mechanical inactivity of the human diaphragm during mechanical ventilation rapidly causes diaphragm atrophy and weakness. However, conclusive evidence for the notion that diaphragm weakness is a direct consequence of mechanical inactivity is lacking. To study the effect of hemidiaphragm paralysis on diaphragm muscle fiber function and structure in humans, biopsies were obtained from the paralyzed hemidiaphragm in eight patients with hemidiaphragm paralysis. All patients had unilateral paralysis of known duration, caused by en bloc resection of the phrenic nerve with a tumor. Furthermore, diaphragm biopsies were obtained from three control subjects. The contractile performance of demembranated muscle fibers was determined, as well as fiber ultrastructure and morphology. Finally, expression of E3 ligases and proteasome activity was determined to evaluate activation of the ubiquitin proteasome pathway. The force-generating capacity, as well as myofibrillar ultrastructure, of diaphragm muscle fibers was preserved up to 8 wk of paralysis. The cross-sectional area of slow fibers was reduced after 2 wk of paralysis; that of fast fibers was preserved up to 8 wk. The expression of the E3 ligases MAFbx and MuRF-1 and proteasome activity was not significantly upregulated in diaphragm fibers following paralysis, not even after 72 and 88 wk of paralysis, at which time marked atrophy of slow and fast diaphragm fibers had occurred. Diaphragm muscle fiber atrophy and weakness following hemidiaphragm paralysis develops slowly and takes months to occur. PMID- 21622848 TI - Inhibition of neutrophil apoptosis by PAI-1. AB - Increased circulating and tissue levels of plasminogen activator inhibitor 1 (PAI 1) are often present in severe inflammatory states associated with neutrophil activation and accumulation and correlate with poor clinical outcome from many of these conditions. The mechanisms by which PAI-1 contributes to inflammation have not been fully delineated. In the present experiments, we found that addition of PAI-1 to neutrophil cultures diminished the rate of spontaneous and TNF-related apoptosis-inducing ligand-induced apoptotic cell death. The effects of PAI-1 on cell viability were associated with activation of antiapoptotic signaling pathways, including upregulation of PKB/Akt, Mcl-1, and Bcl-x(L). Although urokinase-plasminogen activator receptor, lipoprotein receptor-related protein, and vitronectin are primary ligands for PAI-1, these molecules were not involved in mediating its antiapoptotic properties. In contrast, blocking pertussis toxin sensitive G protein-coupled receptors and selective inhibition of phosphatidylinositide 3-kinase reversed the ability of PAI-1 to extend neutrophil viability. The antiapoptotic effects of PAI-1 were also evident under in vivo conditions during LPS-induced acute lung injury, where enhanced apoptosis was present among neutrophils accumulating in the lungs of PAI-1(-/-) compared with PAI-1(+/+) mice. These results demonstrate a novel antiapoptotic role for PAI-1 that may contribute to its participation in neutrophil-associated inflammatory responses. PMID- 21622849 TI - Dysfunctional attitudes and expectancies in deficit syndrome schizophrenia. AB - The deficit syndrome was proposed over 20 years ago as a separate negative symptom syndrome within schizophrenia with a distinct neurobiological pathophysiology and etiology. Recent research, however, has indicated that psychological factors such as negative attitudes and expectancies are significantly associated with the broad spectrum of negative symptoms. Specifically, defeatist beliefs regarding performance mediate between neurocognitive impairment and both negative symptoms and functional outcome. Additionally, asocial beliefs predict asocial behavior and negative expectancies regarding future pleasure are associated with negative symptoms. The present study explored whether these dysfunctional beliefs and negative expectancies might also be a feature of the deficit syndrome. Based on a validated proxy method, 22 deficit and 72 nondeficit patients (from a pool of 139 negative symptom patients) were identified and received a battery of symptom, neurocognitive, and psychological measures. The deficit group scored significantly worse on measures of negative symptoms, insight, emotion recognition, defeatist attitudes, and asocial beliefs but better on measures of depression, anxiety, and distress than the nondeficit group. Moreover, the deficit group showed a trend for higher scores on self-esteem. Based on these findings, we propose a more comprehensive formulation of deficit schizophrenia, characterized by neurobiological factors and a cluster of psychological attributes that lead to withdrawal and protect the self-esteem. Although the patients have apparently opted-out of participation in normal activities, we suggest that a psychological intervention that targets these negative attitudes might improve their functioning and quality of life. PMID- 21622850 TI - Follistatin-like 1 in chronic systolic heart failure: a marker of left ventricular remodeling. AB - BACKGROUND: Follistatin-like 1 (FSTL1) is an extracellular glycoprotein found in human serum. Recent work suggests that FSTL1 is secreted in response to ischemic injuries and that its overexpression is protective in the heart and vasculature. METHODS AND RESULTS: We examined serum FSTL1 levels in patients with chronic heart failure with left ventricular (LV) ejection fraction <40% (n=86). The sample was separated into three tertiles of patients with low, medium, and high FSTL1 levels. Serum FSTL1 was increased 56% above age- and sex-matched healthy controls. Diabetes mellitus, brain natriuretic peptide level, left atrial size, LV posterior wall thickness, LV end-diastolic diameter, and LV mass were significant determinants of FSTL1 serum levels by bivariate analysis. After controlling for significant covariates, FSTL1 levels predicted LV hypertrophy (as measured by LV mass index) by multivariate linear regression analysis (P<0.001). Unadjusted survival analysis demonstrated increased mortality in patients with increasing FSTL1 levels (P=0.09). After adjusting for significant parameters, patients with increased FSTL1 remained at the highest risk of death (hazard ratio, 1.028; 95% CI, 0.98 to 1.78; P=0.26). To determine whether elevated FSTL1 levels may be derived from the myocardium, FSTL1 protein expression was measured in explanted failing (n=18) and nonfailing (n=7) human hearts. LV failing hearts showed 2.5-fold higher FSTL1 protein levels over nonfailing control hearts (P<0.05). CONCLUSIONS: Elevated serum FSTL1 in patients with heart failure was associated with LV hypertrophy. Further studies on the role of FSTL1 as a biomarker in chronic systolic heart failure are warranted. PMID- 21622851 TI - Risk assessment tools for identifying individuals at risk of developing type 2 diabetes. AB - Trials have demonstrated the preventability of type 2 diabetes through lifestyle modifications or drugs in people with impaired glucose tolerance. However, alternative ways of identifying people at risk of developing diabetes are required. Multivariate risk scores have been developed for this purpose. This article examines the evidence for performance of diabetes risk scores in adults by 1) systematically reviewing the literature on available scores and 2) their validation in external populations; and 3) exploring methodological issues surrounding the development, validation, and comparison of risk scores. Risk scores show overall good discriminatory ability in populations for whom they were developed. However, discriminatory performance is more heterogeneous and generally weaker in external populations, which suggests that risk scores may need to be validated within the population in which they are intended to be used. Whether risk scores enable accurate estimation of absolute risk remains unknown; thus, care is needed when using scores to communicate absolute diabetes risk to individuals. Several risk scores predict diabetes risk based on routine noninvasive measures or on data from questionnaires. Biochemical measures, in particular fasting plasma glucose, can improve prediction of such models. On the other hand, usefulness of genetic profiling currently appears limited. PMID- 21622852 TI - Macromolecular assembly of polycystin-2 intracytosolic C-terminal domain. AB - Mutations in PKD2 are responsible for approximately 15% of the autosomal dominant polycystic kidney disease cases. This gene encodes polycystin-2, a calcium permeable cation channel whose C-terminal intracytosolic tail (PC2t) plays an important role in its interaction with a number of different proteins. In the present study, we have comprehensively evaluated the macromolecular assembly of PC2t homooligomer using a series of biophysical and biochemical analyses. Our studies, based on a new delimitation of PC2t, have revealed that it is capable of assembling as a homotetramer independently of any other portion of the molecule. Our data support this tetrameric arrangement in the presence and absence of calcium. Molecular dynamics simulations performed with a modified all-atoms structure-based model supported the PC2t tetrameric assembly, as well as how different populations are disposed in solution. The simulations demonstrated, indeed, that the best-scored structures are the ones compatible with a fourfold oligomeric state. These findings clarify the structural properties of PC2t domain and strongly support a homotetramer assembly of PC2. PMID- 21622853 TI - Spontaneous speciation by ploidy elevation: laboratory synthesis of a new clonal vertebrate. PMID- 21622854 TI - Counterbalancing angiogenic regulatory factors control the rate of cancer progression and survival in a stage-specific manner. AB - Whereas the roles of proangiogenic factors in carcinogenesis are well established, those of endogenous angiogenesis inhibitors (EAIs) remain to be fully elaborated. We investigated the roles of three EAIs during de novo tumorigenesis to further test the angiogenic balance hypothesis, which suggests that blood vessel development in the tumor microenvironment can be governed by a net loss of negative regulators of angiogenesis in addition to the well established principle of up-regulated angiogenesis inducers. In a mouse model of pancreatic neuroendocrine cancer, administration of endostatin, thrombospondin-1, and tumstatin peptides, as well as deletion of their genes, reveal neoplastic stage-specific effects on angiogenesis, tumor progression, and survival, correlating with endothelial expression of their receptors. Deletion of tumstatin and thrombospondin-1 in mice lacking the p53 tumor suppressor gene leads to increased incidence and reduced latency of angiogenic lymphomas associated with diminished overall survival. The results demonstrate that EAIs are part of a balance mechanism regulating tumor angiogenesis, serving as intrinsic microenvironmental barriers to tumorigenesis. PMID- 21622855 TI - Rescue of tumor-infiltrating lymphocytes from activation-induced cell death enhances the antitumor CTL response in CD5-deficient mice. AB - The CD5 coreceptor is expressed on all T cells and on the B1a B cell subset. It is associated with TCR and BCR, and modulates intracellular signals initiated by both Ag receptor complexes. Human CD5 contributes to regulation of the antitumor immune response and susceptibility of specific CTL to activation-induced cell death (AICD) triggered by the tumor. In this study, we compared the T cell response to the B16F10 melanoma engrafted into CD5-deficient and wild-type C57BL/6 mice. Compared with wild-type mice, CD5 knockout animals displayed delayed tumor growth, associated with tumor infiltration by T cell populations exhibiting a more activated phenotype and enhanced antitumor effector functions. However, control of tumor progression in CD5(-/-) mice was transient due to increased AICD of CD8(+) tumor-infiltrating T lymphocytes. Remarkably, in vivo protection of T cells from TCR-mediated apoptosis by an adenovirus engineered to produce soluble Fas resulted in a dramatic reduction in tumor growth. Our data suggest that recruitment of tumor-specific T cells in the tumor microenvironment occurs at early stages of cancer development and that tumor-mediated AICD of tumor-infiltrating T lymphocytes is most likely involved in tumor escape from the immune system. PMID- 21622856 TI - MUC5AC expression through bidirectional communication of Notch and epidermal growth factor receptor pathways. AB - Hyperproduction of goblet cells and mucin in the airway epithelium is an important feature of airway inflammatory diseases. We investigated the involvement of Notch signaling in MUC5AC expression in NCI-H292 cells, a human lung carcinoma cell line. Epidermal growth factor (EGF) stimulated generation of the Notch intracellular domain (NICD) in a RBP-Jkappa-dependent manner. Treatment with gamma-secretase inhibitors L-685,458 or DAPT or introduction of small interfering RNA directed against Notch1 reduced EGF-induced MUC5AC expression. The inhibitory effect of L-685,458 on EGF-induced MUC5AC mRNA and protein expression was also observed in primary human bronchial epithelial cells. Blockage of Notch signaling with L-685,458 or Notch siRNA resulted in a decrease in EGF-induced phosphorylation of ERK. These results suggested that ERK activation is necessary for the regulation of EGF receptor (EGFR)-mediated MUC5AC expression by Notch signaling. Conversely, forced expression of NICD induced both EGFR and ERK phosphorylation with MUC5AC expression even in the absence of EGF. Treatment of the NICD-expressing cells with EGF further augmented ERK phosphorylation in an additive manner. The ERK phosphorylation induced by exogenous NICD was inhibited by treatment with an Ab that antagonizes EGFR activity as well as by inhibitors of EGFR and ERK, implying that Notch signaling induces MUC5AC expression by activating the EGFR pathway. Collectively, these results suggest that MUC5AC expression is regulated by a bidirectional circuit between Notch and EGFR signaling pathways. PMID- 21622857 TI - Mechanical interactions between dendritic cells and T cells correlate with T cell responsiveness. AB - Ag recognition is achieved through the communication across intercellular contacts between T cells and APCs such as dendritic cells (DC). Despite remarkable progress in delineating detailed molecular components at the intercellular contacts, little is known about the functional roles of physical cross-junctional adhesion between T and DC in shaping T cell responses. In addition, the mechanisms underlying sensitivity and specificity of Ag discrimination by T cells at intercellular contacts remain to be elucidated. In this study, we use single-cell force spectroscopy to probe the mechanical interactions between DC and T cells in response to stimulation with a panel of altered peptide ligands. The results show that intercellular interactions of DC-T cell conjugates exhibited different ranges of interaction forces in peptide dependent manners that match the ability of the peptides to activate T cells. Elevated calcium mobilization and IL-2 secretion by T cells were only promoted in response to antigenic peptides that induce strong interaction forces, suggesting that mechanically stable DC-T cell contacts are crucial for driving T cell activation. Strong interactions were not solely dependent on cell-surface molecules such as TCRs and the adhesion molecule LFA-1, but were also controlled by cytoskeletal dynamics and the integrity of membrane lipid rafts. These data provide novel mechanical insights into the effect of Ag affinity on intercellular contacts that align with T cell responsiveness. PMID- 21622858 TI - Reduction of the peripheral blood CD56(bright) NK lymphocyte subset in FTY720 treated multiple sclerosis patients. AB - FTY720 (fingolimod) treatment of multiple sclerosis (MS) results in lymphopenia due to increased recruitment into and decreased egress from secondary lymphoid organs of CCR7(+) lymphocytes. Although absolute numbers of NK lymphocytes were reported as being unaltered in FTY720-treated MS patients (MS-FTY), such analyses did not detect a change in a minor subset. Because expression of CCR7 has been described on CD56(bright) NK cells, a minority population of NK cells, we investigated the effect of FTY720 treatment on the phenotype and function of human NK cells in the peripheral circulation of MS patients. MS-FTY patients displayed a decreased proportion of peripheral CD56(bright)CD62L(+)CCR7(+) NK cells compared with untreated MS and healthy donors. In vitro treatment with FTY720-P increased migration of untreated donor NK cells to CXCL12 while reducing the response to CX3CL1 with similar migration responses seen in NK cells from MS FTY patients. FTY720-P inhibited sphingosine 1-phosphate-directed migration of CD56(bright) and CD56(dim) NK cells subsets from untreated healthy donors. IL-12- and IL-15-stimulated NK cells from MS-FTY patients displayed similar capacity to produce IFN-gamma, TNF, IL-10, and MIP-1alpha cytokines/chemokines compared with NK cells from untreated healthy donors and displayed comparable levels of degranulation in response to K562 tumor cells compared with untreated donors. Subset alterations and function of NK cell populations will need to be considered as part of assessing overall immunosurveillance capacity of patients with MS who will receive sustained FTY720 therapy. PMID- 21622859 TI - High-affinity IgE receptors on dendritic cells exacerbate Th2-dependent inflammation. AB - The IgE-mediated and Th2-dependent late-phase reaction remains a mechanistically enigmatic and daunting element of human allergic inflammation. In this study, we uncover the FcepsilonRI on dendritic cells (DCs) as a key in vivo component of this form of allergy. Because rodent, unlike human, DCs lack FcepsilonRI, this mechanism could be revealed only by using a new transgenic mouse model with human like FcepsilonRI expression on DCs. In the presence of IgE and allergen, FcepsilonRI(+) DCs instructed naive T cells to differentiate into Th2 cells in vitro and boosted allergen-specific Th2 responses and Th2-dependent eosinophilia at the site of allergen exposure in vivo. Thus, FcepsilonRI on DCs drives the cascade of pathogenic reactions linking the initial allergen capture by IgE with subsequent Th2-dominated T cell responses and the development of late-phase allergic tissue inflammation. PMID- 21622860 TI - Cytotoxic CD4+ T cell responses to EBV contrast with CD8 responses in breadth of lytic cycle antigen choice and in lytic cycle recognition. AB - EBV, a B lymphotropic herpesvirus, encodes two immediate early (IE)-, >30 early (E)-, and >30 late (L)-phase proteins during its replication (lytic) cycle. Despite this, lytic Ag-induced CD8 responses are strongly skewed toward IE and a few E proteins only, all expressed before HLA I presentation is blocked in lytically infected cells. For comparison, we examined CD4(+) T cell responses to eight IE, E, or L proteins, screening 14 virus-immune donors to overlapping peptide pools in IFN-gamma ELISPOT assays, and established CD4(+) T cell clones against 12 defined epitopes for target-recognition assays. We found that the lytic Ag-specific CD4(+) T cell response differs radically from its CD8 counterpart in that it is widely distributed across IE, E, and L Ag targets, often with multiple reactivities detectable per donor and with IE, E, or L epitope responses being numerically dominant, and that all CD4(+) T cell clones, whether IE, E, or L epitope-specific, show strong recognition of EBV-transformed B cell lines, despite the lines containing only a small fraction of lytically infected cells. Efficient recognition occurs because lytic Ags are released into the culture and are acquired and processed by neighboring latently infected cells. These findings suggested that lytic Ag-specific CD4 responses are driven by a different route of Ag display than drives CD8 responses and that such CD4 effectors could be therapeutically useful against EBV-driven lymphoproliferative disease lesions, which contain similarly small fractions of EBV-transformed cells entering the lytic cycle. PMID- 21622861 TI - A balance of Bruton's tyrosine kinase and SHIP activation regulates B cell receptor cluster formation by controlling actin remodeling. AB - The activation of the BCR, which initiates B cell activation, is triggered by Ag induced self-aggregation and clustering of receptors at the cell surface. Although Ag-induced actin reorganization is known to be involved in BCR clustering in response to membrane-associated Ag, the underlying mechanism that links actin reorganization to BCR activation remains unknown. In this study, we show that both the stimulatory Bruton's tyrosine kinase (Btk) and the inhibitory SHIP-1 are required for efficient BCR self-aggregation. In Btk-deficient B cells, the magnitude of BCR aggregation into clusters and B cell spreading in response to an Ag-tethered lipid bilayer is drastically reduced, compared with BCR aggregation observed in wild-type B cells. In SHIP-1(-/-) B cells, although surface BCRs aggregate into microclusters, the centripetal movement and growth of BCR clusters are inhibited, and B cell spreading is increased. The persistent BCR microclusters in SHIP-1(-/-) B cells exhibit higher levels of signaling than merged BCR clusters. In contrast to the inhibition of actin remodeling in Btk deficient B cells, actin polymerization, F-actin accumulation, and Wiskott Aldrich symptom protein phosphorylation are enhanced in SHIP-1(-/-) B cells in a Btk-dependent manner. Thus, a balance between positive and negative signaling regulates the spatiotemporal organization of the BCR at the cell surface by controlling actin remodeling, which potentially regulates the signal transduction of the BCR. This study suggests a novel feedback loop between BCR signaling and the actin cytoskeleton. PMID- 21622862 TI - A role for IL-27 in limiting T regulatory cell populations. AB - IL-27 is a cytokine that regulates Th function during autoimmune and pathogen induced immune responses. Although previous studies have shown that regulatory T cells (Tregs) express the IL-27R, and that IL-27 inhibits forkhead box P3 upregulation in vitro, little is known about how IL-27 influences Tregs in vivo. The studies presented in this article show that mice that overexpress IL-27 had decreased Treg frequencies and developed spontaneous inflammation. Although IL-27 did not cause mature Tregs to downregulate forkhead box P3, transgenic overexpression in vivo limited the size of a differentiating Treg population in a bone marrow chimera model, which correlated with reduced production of IL-2, a vital cytokine for Treg maintenance. These data identify an indirect role for IL 27 in shaping the Treg pool. PMID- 21622863 TI - Group X secretory phospholipase A2 enhances TLR4 signaling in macrophages. AB - Secretory phospholipase A(2)s (sPLA(2)) hydrolyze glycerophospholipids to liberate lysophospholipids and free fatty acids. Although group X (GX) sPLA(2) is recognized as the most potent mammalian sPLA(2) in vitro, its precise physiological function(s) remains unclear. We recently reported that GX sPLA(2) suppresses activation of the liver X receptor in macrophages, resulting in reduced expression of liver X receptor-responsive genes including ATP-binding cassette transporters A1 (ABCA1) and G1 (ABCG1), and a consequent decrease in cellular cholesterol efflux and increase in cellular cholesterol content (Shridas et al. 2010. Arterioscler. Thromb. Vasc. Biol. 30: 2014-2021). In this study, we provide evidence that GX sPLA(2) modulates macrophage inflammatory responses by altering cellular cholesterol homeostasis. Transgenic expression or exogenous addition of GX sPLA(2) resulted in a significantly higher induction of TNF-alpha, IL-6, and cyclooxygenase-2 in J774 macrophage-like cells in response to LPS. This effect required GX sPLA(2) catalytic activity, and was abolished in macrophages that lack either TLR4 or MyD88. The hypersensitivity to LPS in cells overexpressing GX sPLA(2) was reversed when cellular free cholesterol was normalized using cyclodextrin. Consistent with results from gain-of-function studies, peritoneal macrophages from GX sPLA(2)-deficient mice exhibited a significantly dampened response to LPS. Plasma concentrations of inflammatory cytokines were significantly lower in GX sPLA(2)-deficient mice compared with wild-type mice after LPS administration. Thus, GX sPLA(2) amplifies signaling through TLR4 by a mechanism that is dependent on its catalytic activity. Our data indicate this effect is mediated through alterations in plasma membrane free cholesterol and lipid raft content. PMID- 21622864 TI - IL-13 antibodies influence IL-13 clearance in humans by modulating scavenger activity of IL-13Ralpha2. AB - Human studies using Abs to two different, nonoverlapping epitopes of IL-13 suggested that epitope specificity can have a clinically significant impact on clearance of IL-13. We propose that Ab modulation of IL-13 interaction with IL 13Ralpha2 underlies this effect. Two Abs were administered to healthy subjects and mild asthmatics in separate dose-ranging studies and allergen-challenge studies. IMA-638 allows IL-13 interaction with IL-13Ralpha1 or IL-13Ralpha2 but blocks recruitment of IL-4Ralpha to the IL-13/IL-13Ralpha1 complex, whereas IMA 026 competes with IL-13 interaction with IL-13Ralpha1 and IL-13Ralpha2. We found ~10-fold higher circulating titer of captured IL-13 in subjects treated with IMA 026 compared with those administered IMA-638. To understand how this difference could be related to epitope, we asked whether either Ab affects IL-13 internalization through cell surface IL-13Ralpha2. Humans inducibly express cell surface IL-13Ralpha2 but lack the soluble form that regulates IL-13 responses in mice. Cells with high IL-13Ralpha2 expression rapidly and efficiently depleted extracellular IL-13, and this activity persisted in the presence of IMA-638 but not IMA-026. The potency and efficiency of this clearance pathway suggest that cell surface IL-13Ralpha2 acts as a scavenger for IL-13. These findings could have important implications for the design and characterization of IL-13 antagonists. PMID- 21622865 TI - Self-tolerance checkpoints in CD4 T cells specific for a peptide derived from the B cell antigen receptor. AB - Linked recognition of Ag by B and T lymphocytes is ensured in part by a state of tolerance acquired by CD4 T cells to germline-encoded sequences within the B cell Ag receptor (BCR). We sought to determine how such tolerance is attained when a peptide from the BCR variable (V) region is expressed by small numbers of B cells as it is in the physiological state. Mixed bone marrow (BM) chimeras were generated using donor BM from mice with B cells that expressed a transgene (Tg) encoded kappa L chain and BM from TCR Tg mice in which the CD4 T cells (CA30) were specific for a Vkappa peptide encoded by the kappaTg. In chimeras where few B cells express the kappaTg, many CA30 cells were deleted in the thymus. However, a substantial fraction survived to the CD4 single-positive stage. Among single positive CA30 thymocytes, few reached maturity and migrated to the periphery. Maturation was strongly associated with, and likely promoted by, expression of an endogenous TCR alpha-chain. CD4(+) CA30 cells that reached peripheral lymphoid tissues were Ag-experienced and anergic, and some developed into regulatory cells. These findings reveal several checkpoints and mechanisms that enforce a state of self-tolerance in developing T cells specific for BCR V region sequences, thus ensuring that T cell help to B cells occurs through linked recognition of foreign Ag. PMID- 21622866 TI - Aborted germinal center reactions and B cell memory by follicular T cells specific for a B cell receptor V region peptide. AB - A fundamental problem in immunoregulation is how CD4(+) T cells react to immunogenic peptides derived from the V region of the BCR that are created by somatic mechanisms, presented in MHC II, and amplified to abundance by B cell clonal expansion during immunity. BCR neo Ags open a potentially dangerous avenue of T cell help in violation of the principle of linked Ag recognition. To analyze this issue, we developed a murine adoptive transfer model using paired donor B cells and CD4 T cells specific for a BCR-derived peptide. BCR peptide-specific T cells aborted ongoing germinal center reactions and impeded the secondary immune response. Instead, they induced the B cells to differentiate into short-lived extrafollicular plasmablasts that secreted modest quantities of Ig. These results uncover an immunoregulatory process that restricts the memory pathway to B cells that communicate with CD4 T cells via exogenous foreign Ag. PMID- 21622867 TI - Forced expression of HLA-DM at the surface of dendritic cells increases loading of synthetic peptides on MHC class II molecules and modulates T cell responses. AB - Adoptive transfer of autologous dendritic cells (DCs) loaded with tumor associated CD4 and CD8 T cell epitopes represents a promising avenue for the immunotherapy of cancer. In an effort to increase the loading of therapeutic synthetic peptides on MHC II molecules, we used a mutant of HLA-DM (DMY) devoid of its lysosomal sorting motif and that accumulates at the cell surface. Transfection of DMY into HLA-DR(+) cells resulted in increased loading of the exogenously supplied HA(307-318) peptide, as well as increased stimulation of HA specific T cells. Also, on transduction in mouse and human DCs, DMY increased loading of HEL(48-61) and of the tumor Ag-derived gp100(174-190) peptides, respectively. Interestingly, expression of DMY at the surface of APCs favored Th1 differentiation over Th2. Finally, we found that DMY(-) and DMY(+) mouse APCs differentially stimulated T cell hybridomas sensitive to the fine conformation of peptide-MHC II complexes. Taken together, our results suggest that the overexpression of HLA-DMY at the plasma membrane of DCs may improve quantitatively, but also qualitatively, the presentation of CD4 T cell epitopes in cellular vaccine therapies for cancer. PMID- 21622868 TI - Cutting edge: an NK cell-independent role for Slamf4 in controlling humoral autoimmunity. AB - Several genes within a syntenic region of human and mouse chromosome 1 are associated with predisposition to systemic lupus erythematosus. Analyses of lupus prone congenic mice have pointed to an important role for the signaling lymphocyte activation molecule family (slamf)6 surface receptor in lupus pathogenesis. In this article, we demonstrate that a second member of the Slamf gene family, Slamf4 (Cd244), contributes to lupus-related autoimmunity. B6.Slamf4(-/-) mice spontaneously develop activated CD4 T cells and B cells and increased numbers of T follicular helper cells and a proportion develop autoantibodies to nuclear Ags. B6.Slamf4(-/-) mice also exhibit markedly increased autoantibody production in the B6.C-H-2bm12/KhEg -> B6 transfer model of lupus. Although slamf4 function is best characterized in NK cells, the enhanced humoral autoimmunity of B6.Slamf4(-/-) mice is NK cell independent, as judged by depletion studies. Taken together, our findings reveal that slamf4 has an NK cell-independent negative regulatory role in the pathogenesis of lupus a normally non-autoimmune prone genetic background. PMID- 21622869 TI - Serum amyloid A activates the NLRP3 inflammasome and promotes Th17 allergic asthma in mice. AB - IL-1beta is a cytokine critical to several inflammatory diseases in which pathogenic Th17 responses are implicated. Activation of the NLRP3 inflammasome by microbial and environmental stimuli can enable the caspase-1-dependent processing and secretion of IL-1beta. The acute-phase protein serum amyloid A (SAA) is highly induced during inflammatory responses, wherein it participates in systemic modulation of innate and adaptive immune responses. Elevated levels of IL-1beta, SAA, and IL-17 are present in subjects with severe allergic asthma, yet the mechanistic relationship among these mediators has yet to be identified. In this study, we demonstrate that Saa3 is expressed in the lungs of mice exposed to several mixed Th2/Th17-polarizing allergic sensitization regimens. SAA instillation into the lungs elicits robust TLR2-, MyD88-, and IL-1-dependent pulmonary neutrophilic inflammation. Furthermore, SAA drives production of IL 1alpha, IL-1beta, IL-6, IL-23, and PGE(2), causes dendritic cell (DC) maturation, and requires TLR2, MyD88, and the NLRP3 inflammasome for secretion of IL-1beta by DCs and macrophages. CD4(+) T cells polyclonally stimulated in the presence of conditioned media from SAA-exposed DCs produced IL-17, and the capacity of polyclonally stimulated splenocytes to secrete IL-17 is dependent upon IL-1, TLR2, and the NLRP3 inflammasome. Additionally, in a model of allergic airway inflammation, administration of SAA to the lungs functions as an adjuvant to sensitize mice to inhaled OVA, resulting in leukocyte influx after Ag challenge and a predominance of IL-17 production from restimulated splenocytes that is dependent upon IL-1R signaling. PMID- 21622871 TI - Strategic supplementation of calcium salts of polyunsaturated fatty acids to enhance reproductive performance of Bos indicus beef cows. AB - Five experiments evaluated the effects of supplemental Ca salts of PUFA on reproductive function of Bos indicus beef cows. In Exp. 1, nonlactating and multiparous grazing cows (n = 51) were assigned to receive (as-fed basis) 0.1 kg of a protein-mineral mix + 0.1 kg of ground corn per cow/d, in addition to 0.1 kg per cow/d of 1) Ca salts of PUFA (PF), 2) Ca salts of SFA (SF), or 3) kaolin (control). Treatments were offered from d 0 to 20 of the estrous cycle. No treatment effects were detected on serum progesterone concentrations (P = 0.83), day of luteolysis (P = 0.86), or incidence of short cycles (P = 0.84). In Exp. 2, nonlactating and multiparous grazing cows (n = 43) were assigned to receive PF, SF, or control from d 0 to 8 of the estrous cycle. On d 6, all cows received (intramuscularly) 25 mg of PGF(2alpha). No treatment effects were detected on serum progesterone concentrations on d 6 (P = 0.37), and incidence (P = 0.67) or estimated time of luteolysis (P = 0.44). In Exp. 3, twenty-seven lactating and multiparous grazing cows, approximately 30 to 40 d postpartum, were assigned to receive PF or control for 10 d beginning at the first postpartum ovulation. No treatment effects were detected (P = 0.85) on incidence of short cycles. In Exp. 4, lactating and multiparous grazing cows (n = 1,454), approximately 40 to 60 d postpartum, were assigned to receive 1 of the 7 treatments for 28 d after timed AI (TAI; d 0): 1) control from d 0 to 28, 2) SF from d 0 to 14 and then control, 3) PF from d 0 to 14 and then control, 4) SF from d 0 to 21 and then control, 5) PF from d 0 to 21 and then control, 6) SF from d 0 to 28, and 7) PF from d 0 to 28. Cows receiving PF for more than 21 d after TAI had greater (P < 0.01) pregnancy to TAI compared with all other treatments combined (50.4 vs. 42.4%, respectively). In Exp. 5, lactating and multiparous grazing cows (n = 501), approximately 40 to 60 d postpartum, were assigned to receive 1 of the 4 treatments for 21 d after TAI (d 0): 1) PF from d 0 to 14 and then control, 2) control from d 0 to 6 and then PF, 3) control from d 0 to 13 and then PF, and 4) PF from d 0 to 21. Cows receiving PF after d 14 of the experiment had greater (P = 0.02) pregnancy to TAI compared with cows not receiving PF during the same period (46.8 vs. 33.1%, respectively). In summary, supplemental Ca salts of PUFA during the expected time of luteolysis increased pregnancy to TAI in beef cows. PMID- 21622872 TI - Protein-carbohydrate interactions between Lactobacillus salivarius and pig mucins. AB - Adherence to the gastrointestinal tract is a key element desirable for many of the proposed beneficial health effects of probiotic bacteria. The aims of this study were to determine the amounts of adhesion of 3 Lactobacillus salivarius strains (Lb6, Lb9, and Lb10) to porcine small intestinal mucins and to determine whether adhesion is a function of lectin-like activities. Dot and Western blot assays were performed to investigate bacterial adhesion. Several carbohydrates and glycoproteins were evaluated to determine whether they interfered with adhesion of the Lactobacillus strains to intestinal mucins and to determine whether they had lectin-like activities. The Lb9 and Lb10 strains had greater association with piglet mucins than did those from 22- to 24-wk-old finishing pigs (P = 0.021 and 0.037, respectively), whereas the Lb6 strain adhered to both (P = 0.138). Western blot assays showed that bacterial adhesion detected piglet mucosa from the duodenum, jejunum, and ileum. In finishing pigs, the adhesion was variable throughout the gastrointestinal tract. Galactose and mannose diminished the interaction of the Lb9 and Lb10 strains in intestinal mucosa (P = 0.028 and 0.026, respectively), whereas pig gastric mucin reduced the adhesion of the Lb6 strain (P = 0.013). Adhesion of the Lb9 and Lb10 strains to intestinal mucosa was less after protease treatment (P = 0.023 and 0.018, respectively), which indicates that proteins are needed for the Lb9 and Lb10 strains to recognize mucin. The Lb6 strain also demonstrated diminished adhesion after periodate treatment (P = 0.038). From these results, we suggest that the nature of the bacterial lectin-like substance is a surface protein that loosely binds to the bacterial cell surface. All the tested strains adhered to specific targets in the small intestinal mucosa of piglets, and the bacteria had lectin-like proteins involved in this adhesion. PMID- 21622873 TI - Nitrous oxide by itself is insufficient to relieve pain due to castration in piglets. AB - Surgical castration is performed on most male piglets in the United States. However, castration is painful and analgesics have been considered to relieve pain. Inhalant gases with analgesic properties allow for a fast induction, have short-term and reversible effects, and are a needle-free option. Nitrous oxide (N(2)O; "laughing gas") has been widely used in human surgery and dental offices as an analgesic, sedative, and anxiolytic drug, yet N(2)O has not been thoroughly investigated for use in farm animals. We hypothesized that the analgesic effect of N(2)O could reduce the pain experienced by piglets during or immediately after castration. Twenty-four male piglets, from 12 litters, were castrated at 3 d of age. One piglet received N(2)O and a littermate received air as a control. After 150 s of exposure to the gas, castration was performed while the piglet remained exposed to the gas. Agitation scores and total vocalization length were recorded during castration. Behavioral observations were continued for 3 d postcastration by using a 5-min scan-sampling method for 4 h the first morning and for 2-h periods in the morning and afternoon of each day thereafter. Body weight gain was measured on the day before castration, at 3 d postcastration, and at weaning. Data were analyzed using a mixed model in SAS (Cary, NC). Nitrous oxide successfully induced anesthesia in all N(2)O piglets, as validated by a skin pinch test and the loss of the palpebral reflex. Total vocalization length was shorter in piglets receiving N(2)O during the induction phase (P = 0.003) but was not different during castration itself because piglets receiving N(2)O awoke and vocalized as much as control piglets (P = 0.87). Agitation scores during the whole procedure were reduced in piglets receiving N(2)O in both frequency (P = 0.005) and intensity (P = 0.026). For 2 h after castration, piglets receiving N(2)O displayed less huddling behavior than did control piglets (P = 0.01). Over the 3 d, piglets receiving N(2)O performed more tail wagging (P = 0.02) and tended to show fewer sleep spasms (P = 0.06) than did control piglets. Piglets given N(2)O tended to have a reduced growth rate compared with control piglets at 3 d postcastration and at weaning (P = 0.05 and P = 0.06, respectively). Nitrous oxide was effective in inducing anesthesia in neonatal piglets during handling. Nonetheless, its analgesic effects appeared insufficient in preventing castration induced pain. PMID- 21622874 TI - Selection for ovulation rate in rabbits: genetic parameters, direct response, and correlated response on litter size. AB - The aim of this work was to evaluate the response to 10 generations of selection for ovulation rate. Selection was based on the phenotypic value of ovulation rate, estimated at d 12 of the second gestation by laparoscopy. Selection pressure was approximately 30%. Line size was approximately 20 males and 80 females per generation. Traits recorded were ovulation rate at the second gestation, estimated by laparoscopy as the number of corpora lutea in both ovaries; ovulation rate at the last gestation, estimated postmortem; ovulation rate, analyzed as a single trait including ovulation rate at the second gestation and ovulation rate at the last gestation; right and left ovulation rates; ovulatory difference, estimated as the difference between the right and left ovulation rates; litter size, estimated as the total number of kits born and the number of kits born alive, both recorded at each parity. Totals of 1,477 and 3,031 records from 900 females were used to analyze ovulation rate and litter size, respectively, whereas 1,471 records were used to analyze ovulatory difference, right ovulation rate, and left ovulation rate. Data were analyzed using Bayesian methodology. Heritabilities of ovulation rate, litter size, number of kits born alive, right ovulation rate, left ovulation rate, and ovulatory difference were 0.16, 0.09, 0.08, 0.09, 0.04 and 0.03, respectively. Phenotypic correlations of ovulation rate with litter size, number of kits born alive, and ovulatory difference were 0.09, 0.01, and 0.14, respectively. Genetic correlations of ovulation rate with litter size and with number of kits born alive were estimated with low accuracy, and there was not much evidence for the sign of the correlation. The genetic correlation between ovulation rate and ovulatory difference was positive (P = 0.91). In 10 generations of selection, ovulation rate increased in 1.32 oocytes, with most of the response taking place in the right ovary (1.06 oocytes), but there was no correlated response on litter size (-0.15 kits). In summary, the direct response to selection for ovulation rate was relevant, but it did not modify litter size because of an increase in prenatal mortality. PMID- 21622875 TI - Impact of dietary energy level and ractopamine on growth performance, carcass characteristics, and meat quality of finishing pigs. AB - A total of 54 finishing barrows (initial BW = 99.8 +/- 5.1 kg; PIC C22 * 337) reared in individual pens were allotted to 1 of 6 dietary treatments in a 2 * 3 factorial arrangement of treatments with 2 levels of ractopamine (0 and 7.4 mg/kg) and 3 levels of dietary energy (high, 3,537; medium, 3,369; and low, 3,317 kcal of ME/kg) to determine the effects of dietary ractopamine and various energy levels on growth performance, carcass characteristics, and meat quality of finishing pigs. High-energy diets were corn-soybean-meal-based with 4% added fat; medium-energy diets were corn-soybean meal based with 0.5% added fat; and low energy diets were corn-soybean meal based with 0.5% added fat and 15% wheat middlings. Diets within each ractopamine level were formulated to contain the same standardized ileal digestible Lys:ME (0 mg/kg, 1.82; and 7.4 mg/kg, 2.65 g/Mcal of ME). Individual pig BW and feed disappearance were recorded at the beginning and conclusion (d 21) of the study. On d 21, pigs were slaughtered for determination of carcass characteristics and meat quality. No ractopamine * energy level interactions (P > 0.10) were observed for any response criteria. Final BW (125.2 vs. 121.1 kg), ADG (1.2 vs. 1.0 kg/d), and G:F (0.31 vs. 0.40) were improved (P < 0.001) with feeding of ractopamine diets. Feeding of the low energy diet reduced (P = 0.001) final BW and ADG compared with the high- and medium-energy diets. Gain:feed was reduced (P = 0.005) when the medium-energy diets were fed compared with the high-energy diets. Additionally, G:F was reduced (P = 0.002) when the low-energy diets were compared with the high- and medium energy diets. Feeding ractopamine diets increased (P < 0.05) HCW (93.6 vs. 89.9 kg) and LM area (51.2 vs. 44.2 cm(2)). The LM pH decline was reduced (P <= 0.05) by feeding ractopamine diets. The feeding of low-energy diets reduced (P = 0.001) HCW when compared with the high- and medium-energy diets and reduced (P = 0.024) 10th-rib backfat when compared with the high- and medium-energy diet. These data indicate that feeding ractopamine diets improved growth performance and carcass characteristics, while having little or no detrimental effect on meat quality. Reductions in energy content of the diet by adding 15% wheat middlings resulted in impaired ADG, G:F, and 10th-rib backfat. There were no ractopamine * energy level interactions in this trial, which indicates that the improvements resulting from feeding ractopamine were present regardless of the dietary energy levels. PMID- 21622876 TI - Effect of the FecX(R) polymorphism in the bone morphogenetic protein 15 gene on natural or equine chorionic gonadotropin-induced ovulation rate and litter size in Rasa Aragonesa ewes and implications for on-farm application. AB - A new mutation in the bone morphogenetic protein 15 (BMP15) gene (FecX(R) allele) causing increased prolificacy in heterozygous (R+) and sterility in homozygous ewes has been recently described in Rasa Aragonesa, a low-prolificacy Mediterranean breed. The current study determined, first, the effect of this polymorphism on natural and eCG-induced ovulation rate (OR) and the effect of eCG dose on reproductive performance; and second, its effect on prolificacy and its interaction with progestagen + eCG treatment on farms, which have not been reported to date. The FecX(R) allele increased OR by 0.44 and 0.63 ovulations in young (n = 91) and adult (n = 84) R+ ewes, respectively (both, P < 0.01), increments less than reported in prolific breeds carrying other mutations in BMP15. When the standard dose of eCG used on farms (480 IU) was applied to R+ ewes (n = 36), an extremely high OR (3.95) was recorded, which was accompanied by greater partial failure of multiple ovulations (PFMO). On the contrary, OR using 240 IU in R+ ewes (2.90; n = 35) was similar to 480 IU in wildtype (++) ewes (2.82; n = 48; both P < 0.01 when compared with 480 IU in R+ ewes). No differences were found in the birth weight of the offspring between R+ and ++ eCG stimulated ewes within the same litter size. To validate the genealogy identification on farms, PCR genotyping was carried out in 1,667 ewes from 4 elite flocks, resulting in a negligible misclassification of R+ ewes, which demonstrated that identification by genealogy is a reliable tool to identify FecX(R) ewes within the breeding program. In recorded farms, the natural litter size of ++ ewes (1.34, n = 599,160 lambing records) was increased due to the FecX(R) allele by 0.35 lambs (P < 0.0001, n = 6,593 lambing records). A similar increase (0.30) was observed when comparing ++ and R+ ewes treated with 480 IU of eCG (P < 0.0001, n = 62,055 and n = 866, respectively). When applying 480 IU of eCG to R+ ewes, the increase in prolificacy was only due to increased percentages of triplets (P < 0.001) and quadruplets (P < 0.0001), but not of twin births. In conclusion, the favorable reproductive performance of R+ ewes, with 0.63 extra ovulations and 0.35 extra lambs per lambing ewe, is responsible for the increased interest in the use of this polymorphism. Nevertheless, care must be taken in the application of eCG to R+ ewes, with the current results showing that the standard dose increases prolificacy by only increasing triple and higher-order births. PMID- 21622877 TI - Genetic and phenotypic parameter estimates for feed intake and other traits in growing beef cattle, and opportunities for selection. AB - Growth, feed intake, and temperament indicator data, collected over 5 yr on a total of 1,141 to 1,183 mixed-breed steers, were used to estimate genetic and phenotypic parameters. All steers had a portion of Hereford, Angus, or both as well as varying percentages of Simmental, Charolais, Limousin, Gelbvieh, Red Angus, and MARC III composite. Because the steers were slaughtered on various dates each year and the animals thus varied in days on feed, BW and feed data were adjusted to a 140-d feeding period basis. Adjustment of measures of feed efficiency [G:F or residual feed intake (RFI), intake adjusted for metabolic body size, and BW gain] for body fatness recorded at slaughter had little effect on the results of analyses. Average daily gain was less heritable (0.26) than was midtest BW (MBW; 0.35). Measures of feed intake had greater estimates of heritability, with 140-d DMI at 0.40 and RFI at 0.52; the heritability estimate for G:F was 0.27. Flight speed (FS), as an indicator of temperament, had an estimated heritability of 0.34 and a repeatability of 0.63. As expected, a strong genetic (0.86) correlation was estimated between ADG and MBW; genetic correlations were less strong between DMI and ADG or MBW (0.56 and 0.71). Residual feed intake and DMI had a genetic correlation of 0.66. Indexes for phenotypic RFI and genotypically restricted RFI (no correlation with BW gain) were compared with simple economic indexes incorporating feed intake and growth to elucidate expected selection responses under different criteria. In general, few breed differences were detected across the various measurements. Heterosis contributed to greater DMI, RFI, and MBW, but it did not significantly affect ADG, G:F, or FS. Balancing output (growth) with input costs (feed) is needed in practicing selection, and FS would not be recommended as an indicator trait for selection to change feed efficiency. An index including BW gain and RFI produced the best economic outcome. PMID- 21622878 TI - Fatty acid profiles and iodine value correlations between 4 carcass fat depots from pigs fed varied combinations of ractopamine and energy. AB - A total of 54 finishing barrows (initial BW = 99.8 +/- 5.1 kg; PIC C22 * 337) reared in individual pens were allotted to 1 of 6 dietary treatments in a 2 * 3 factorial arrangement of treatments with 2 levels of ractopamine (0 and 7.4 mg/kg) and 3 levels of dietary energy (high: 3,537, medium: 3,369, and low: 3,317 kcal/kg of ME) to determine the effects of feeding ractopamine and various dietary energy levels on the fatty acid profile of 4 carcass fat depots (jowl, belly, subcutaneous loin, and intramuscular) and the predictive relationships of calculated iodine value (IV) between these 4 fat depots. Carcasses were sampled for fat tissues at the anterior tip of the jowl, posterior to the sternum on the belly edge, three-quarters the distance around the LM (subcutaneous fat; SC), and within the LM (intramuscular fat; IMF). Feeding ractopamine diets reduced (P < 0.05) total SFA in SC and IMF and increased (P = 0.04) total MUFA in SC. Also, feeding ractopamine diets increased (P < 0.01) the IV of IMF. Total MUFA of belly fat was reduced (P < 0.05) when the low-energy diet was fed compared with the high-energy diet. Jowl fat total MUFA was reduced (P < 0.05) and total PUFA was increased (P < 0.05) when the medium-energy diet was fed compared with the high- and low-energy diets. Iodine values, independent of treatment, were 60.97, 64.51, 55.59, and 58.26 for belly, jowl, IMF, and SC fat depots, respectively. The IV correlations within fat depots were not consistent across dietary treatments because of the effect of treatments on carcass fatty acid characteristics. Feeding ractopamine diets shifted the fatty acid profile from SFA to MUFA in the SC depot. Feeding ractopamine diets did not change belly fat profiles, thus avoiding the potential negative effect of softening belly fat, which is detrimental to processing value. The IV of one fat depot may not be a good indication of IV of other fat depots because of weak correlation coefficients and the apparent influence of dietary treatment. PMID- 21622879 TI - Valine partitioning and kinetics between the gastrointestinal tract and hind limbs in lambs with an adult Trichostrongylus colubriformis burden. AB - Intestinal parasitic infection increases the demand for AA because of increased protein synthesis in the intestine and increased luminal losses of AA, and these increased demands may be supported by increased mobilization of AA from the skeletal muscles. Two experiments were conducted to determine the effects of parasitic infection on valine kinetics within the gastrointestinal tract and hind limbs of lambs fed fresh forages. On d 1, lambs were given 6,000 stage-3 Trichostrongylus colubriformis larvae per day for 6 d (n = 6) or kept as parasite free controls (n = 6) and fed fresh lucerne (Medicago sativa; Exp. 1) or fresh sulla (Hedysarum coronarium; Exp. 2). On d 48, valine kinetics within the mesenteric- (MDV) and portal-drained viscera (PDV) and hind limbs were obtained by carrying out concurrent infusions of para-amminohippuric acid into the mesenteric vein and indocyanin green into the abdominal aorta (for blood flow), and [3,4-(3)H]valine into the jugular vein and [1-(13)C]valine into the abomasum for 8 h (for kinetics). During the infusions, blood was collected from the mesenteric and portal veins and from the mesenteric artery and vena cava, and plasma was harvested. After the 8-h infusion, lambs were euthanized, ileal digesta were collected, and tissues were sampled from the intestine and muscle (biceps femoris). Tissues, digesta, and plasma were analyzed for valine concentration, specific radioactivity, and isotopic enrichment. In both experiments, intestinal worm burdens on d 48 were greater in parasitized lambs (P = 0.0001 and 0.003). In Exp. 1, parasitic infection increased (P = 0.03) the total valine irreversible loss rate (ILR) in the MDV and PDV. In Exp. 2, luminal ILR of valine in the MDV was reduced (P = 0.01); however, ILR of valine in the PDV was unaffected. Despite these changes within the MDV and PDV, parasitic infection did not affect the ILR of valine within the hind limbs, and valine transport rates were largely unchanged. We suggest that the increased mobilization of AA from the hind limbs that might have occurred in the early phase of inflammation was no longer required when the parasitic infection was established. The MDV and PDV data may indicate that the non-MDV parts of the PDV play an important role in this adaptation, which warrants further study. PMID- 21622880 TI - Mannan oligosaccharide modulates gene expression profile in pigs experimentally infected with porcine reproductive and respiratory syndrome virus. AB - This study characterized gene expression in peripheral blood mononuclear cells (PBMC) and bronchoalveolar lavage fluid (BALF) cells from control- or mannan oligosaccharide (MOS)-fed pigs with or without porcine reproductive and respiratory syndrome virus (PRRSV) at d 7 postinfection (PI). Weaned pigs (3 wk old) fed 0 or 0.2% MOS (Bio-Mos) diets were intranasally inoculated with PRRSV or a sterile medium at 5 wk of age. Total RNA (3 pigs/treatment) was extracted from cells. Double-stranded cDNA was amplified, labeled, and further hybridized to the Affymetrix GeneChip Porcine Genome Array consisting of 23,937 probe sets representing 20,201 genes. Microarray data were analyzed in R using packages from the Bioconductor project. Differential gene expression was tested by fitting a mixed linear model equivalent to a 2 * 2 factorial ANOVA using the limma package. Dietary MOS and PRRSV changed the expression of thousands of probe sets in PBMC and BALF cells (P < 0.05). The MOS * PRRSV interaction altered the expression of more nonimmune probe sets in PBMC (977 up, 1,128 down) than in BALF cells (117 up, 78 down). The MOS * PRRSV interaction (P < 0.05) for immune probe sets in PBMC affected genes encoding key inflammatory mediators. In uninfected pigs, gene expression of IL-1alpha, IL-6, myeloid differentiation factor 88, Toll-like receptor (TLR) 4, major histocompatibility complex (MHC) II, and dead box polypeptide 58 increased in PBMC of MOS-fed pigs (P < 0.05). This suggests that MOS enhances disease resistance in pigs and supports the fact that MOS induced a rapid increase in leukocytes at d 3 and 7 PI. Within infected pigs, however, MOS reduced the expression of IL-1beta, IL-6, IL-8, macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, monocyte chemotactic protein (MCP)-1, and TLR4 genes in PBMC (P < 0.05). This finding may explain why fever was ameliorated in infected pigs fed MOS by d 7 PI. The expression of IL-1beta, IL-6, MIP-1beta, MCP-1, and TLR4 genes was confirmed by quantitative real-time reverse-transcription PCR. In BALF cells of infected pigs, MOS reduced the gene expression of TLR4, MHCII, and molecules associated with the complement system, but increased the gene expression of MHCI. In short, MOS regulated the expression of nonimmune and immune genes in pig leukocytes, perhaps providing benefits by enhancing the immune responses of the pigs to an infection, while preventing overstimulation of the immune system. PMID- 21622881 TI - Grass silage intake, rumen and blood variables, ultrasonic and body measurements, feeding behavior, and activity in pregnant beef heifers differing in phenotypic residual feed intake. AB - The objectives of this study were to quantify the phenotypic variation in residual feed intake (RFI) in pregnant beef heifers offered a grass silage diet and to characterize their productivity. Seventy-three pregnant (mean gestation d 198, SD = 27 d) Simmental and Simmental * Holstein-Friesian heifers (mean initial BW 548, SD = 47.5 kg) were offered grass silage ad libitum. Heifer DMI, BW, BCS, skeletal measurements, ultrasonic fat and muscle depth, visual muscularity score, rumen fermentation, total tract digestibility, blood metabolite and hematology variables, feeding, and activity behavior were measured during an 84-d feed intake study. After parturition calf birth weight, calving difficulty, cow serum IgG, hematology variables, and calf humoral immune status were measured. In a subset of cows (n = 28), DMI, milk yield and various body composition variables were also measured approximately 3 wk postpartum. Phenotypic RFI was calculated for each animal as the difference between actual DMI and expected DMI. Expected DMI was computed for each animal by regressing average daily DMI on conceptus adjusted mean BW(0.75) and conceptus-adjusted ADG over an 84-d period. Within breed, heifers were ranked by RFI into low (efficient), medium, and high (inefficient) groups by dividing them into thirds. Heifers with high RFI had 8.8 and 17.1% greater (P < 0.001) DMI than medium and low RFI groups, respectively. The RFI groups did not differ in ADG or BW (P > 0.05). Residual feed intake was positively correlated with DMI (r = 0.85) but not with feed conversion ratio, ADG, or BW. The RFI groups did not differ (P > 0.05) in skeletal size, BCS, ultrasonic fat depth, total tract digestibility, calf birth weight, calving difficulty, serum IgG concentrations, or milk yield. Visual muscularity scores, initial test and postpartum ultrasonic muscle depth were negatively correlated with RFI (P < 0.05). Including mean ultrasonic muscle depth into the base RFI regression model increased its R(2) (0.29 to 0.38). Pearson rank correlation between RFI and muscle-adjusted RFI was 0.93. The results show that efficient RFI heifers consumed less feed without any compromise in growth, body composition, or maternal traits measured. PMID- 21622882 TI - Effects of manipulation of the caspase system on myofibrillar protein degradation in vitro. AB - Apoptosis via the intrinsic caspase 9 pathway can be induced by oxidative stressors hydrogen peroxide (H2O2) and N-(4 hydroxyphenol) rentinamide (fenretinide), a synthetic retinoid. Accelerated muscle atrophy and proteolysis in muscle-wasting conditions have been linked to oxidative stress and activated protease systems. Therefore, the hypothesis of this study was that proteolysis of myofibrillar proteins could be manipulated through the induction or inhibition of the caspase system. After slaughter, LM and supraspinatus muscles from callipyge (n = 5) and normal (n = 3) lambs were excised, finely diced, and incubated with treatment buffers containing oxidative stressors fenretinide or H2O2, recombinant caspase 3, caspase-specific inhibitor N-acetyl-Asp-Glu-Val-Asp-CHO (DEVD), or control solution. Muscle samples were incubated for 1, 2, 7, and 21 d at 4 degrees C. Activation of the initiator caspase, caspase 9, and myofibrillar protein degradation was determined by SDS-PAGE and Western blotting. Results showed that fenretinide, H2O2, and recombinant caspase 3 increased (P < 0.05) proteolysis of myofibril proteins, whereas DEVD inhibited degradation (P < 0.05). Proteolysis of myofibrillar proteins increased with incubation time (P < 0.0001), and incubation time * treatment interactions (P < 0.05) indicated that the treatment effects did not all occur at the same rate. This study has shown that manipulation of the caspase system through induction or inhibition of activity can affect degradation of myofibrillar proteins, providing further evidence that the caspase system could be involved in postmortem proteolysis and tenderization. However, these stimulated changes were not sufficient to overcome the lack of proteolysis that is characteristic of muscle from callipyge lambs. PMID- 21622883 TI - Protein and glucogenic precursor supplementation: a nutritional strategy to increase reproductive and economic output. AB - Reproductive performance in young beef cows is often compromised due to a mismatch of physiological demands and suboptimal environmental conditions. Studies conducted at the Corona Range and Livestock Research Center from 2000 to 2007 evaluated 3 postpartum supplement strategies that varied in the amount of glucogenic potential (GP) supplied. Reproductive variables, milk production, and serum metabolites were used to assess supplement effectiveness and economics associated with 2- and 3-yr-old beef cows (n = 379) grazing native range. Supplements were individually fed twice/week at 1,135 g/d (2003 to 2004) or 908 g/d (all other years) and provided 1) 327 g of CP, 109 to 118 g of RUP (CON); 2) 327 to 341 g of CP, 142 to 157 g of RUP (RUP); or 3) 327 g of CP, 151 to 173 g of RUP + 40 to 100 g of propionate salt (PS; RUP+PS). Ultimately, total GP for CON, RUP, and RUP+PS was 44 to 47, 57 to 70, and 93 to 141 g, respectively. Blood samples were collected once/week (2000) or twice/week (2001 to 2007) for progesterone analysis to estimate days to resumption of estrus. Cows were exposed to bulls for 60 d or less, and pregnancy was confirmed by rectal palpation at weaning. Days to resumption of estrus after calving decreased linearly (P = 0.02), resulting in an increased pregnancy rate (P = 0.03) with increasing GP. Milk production exhibited a quadratic (P = 0.04) response to increasing GP, with cows fed RUP producing the most amount of milk. However, a linear decrease (P = 0.07) in days from BW nadir to estrus was found with increasing GP. Total kilograms of calf weaned per cow exposed for the supplemental year and subsequent year was increased linearly (P = 0.07) with increased GP. The improvement in pregnancy rate by supplementing RUP+PS resulted in an increase in total revenue of 18% compared with CON-fed cows and 9.5% compared with RUP-fed cows in the subsequent year after supplementation. These data suggest feeding young cows additional GP in the form of PS allows for partitioning of nutrients away from milk production and toward reproduction, allowing for increased profitability by increasing pregnancy rates and decreasing days to resumption of estrus. PMID- 21622884 TI - Protein supplementation of ruminants consuming low-quality cool- or warm-season forage: differences in intake and digestibility. AB - An in situ study (Exp. 1) using 4 ruminally cannulated steers (343 +/- 11 kg of BW) in a completely randomized design was used to compare ruminal degradation characteristics of low-quality cool-season (C3; Kentucky bluegrass straw; Poa pratensis; 6.3% CP; DM basis) and warm-season (C4; tallgrass prairie; 5.7% CP; DM basis) forage. Four ruminally cannulated steers (252 +/- 8 kg of BW; Exp. 2) and 4 wethers (38 +/- 1 kg of BW; Exp. 3) were used in two 2 * 2 factorial arrangements of treatments to determine the influence of supplemental CP (CPSupp; soybean meal; 0.09 and 0.19% of BW, CP basis, for steers and lambs, respectively) on nutrient intake and digestion of C3 and C4 forages. Steers and wethers were allotted to separate 4 * 4 Latin squares that ran simultaneously with 20-d periods. In Exp. 1, C3 had a greater A fraction (fraction of total pool disappearing at a rate too rapid to measure) and effective degradability of DM and NDF compared with C4 (P < 0.01). In addition, C3 had a greater (P < 0.01) A fraction and effective degradability of N, whereas the C fraction (fraction of total pool unavailable in the rumen) was less (P < 0.01) than those for C4. Consequently, RDP accounted for 84.7% of total CP in C3 as compared with 66% for C4 (P < 0.01). In Exp. 2, a CPSupp * forage interaction (P < 0.01) was noted for forage and total DMI, with CPSupp increasing intake of C4 by 47% and intake of C3 forage by only 7%. Dry matter digestibility responded similarly, with a CPSupp * forage interaction (P = 0.05; CPSupp increased digestibility by 21% with C4 and by 9% with C3 forage). In addition, CPSupp * forage interactions were noted for ruminal liquid retention time (P = 0.02; CPSupp decreased retention by 3.6 h with C4 and by only 0.6 h with C3 forage) and particulate passage rate (P = 0.02; CPSupp increased passage by 46% with C4 and by 10% with C3 forage). As in Exp. 2, a CPSupp * forage interaction (P = 0.01; CPSupp increased digestibility by 18% with C4 and by 7% with C3 forage) was observed with DM digestibility in Exp. 3. In contrast, only N balance (P < 0.01) and N digestibility (P < 0.01) were affected by CPSupp. These data suggest that intake and digestion of low-quality C3 and C4 forages by ruminants are not similar and, more important, that the physiological response of ruminants to protein supplementation of low-quality forage is dependent on forage type. PMID- 21622885 TI - Maternal nutrition during pregnancy influences offspring wool production and wool follicle development. AB - The effects of maternal nutrition on offspring wool production (quality and quantity) were evaluated. Primiparous Rambouillet ewes (n = 84) were randomly allocated to 1 of 6 treatments in a 2 * 3 factorial design. Selenium treatment [adequate Se (ASe, 9.5 MUg/kg of BW) vs. high Se (HSe, 81.8 MUg/kg of BW)] was initiated at breeding, and maternal nutritional intake [control (CON, 100% of requirements) vs. restricted (60% of CON) vs. overfed (140% of CON)] was initiated at d 50 of gestation. Lamb birth weight was recorded at delivery, and all lambs were placed on the same diet immediately after birth to determine the effects of prenatal nutrition on postnatal wool production and follicle development. At 180 +/- 2.2 d of age, lambs were necropsied and pelt weights were recorded. Wool samples were collected from the side and britch areas, whereas skin samples were collected from the side of each lamb only. Although Se status did not influence side staple length in males, female lambs born from ewes on the ASe treatment had a shorter staple length (P < 0.05) when compared with females from ewes on the HSe treatment. Maternal nutritional intake and Se status did not influence (P >= 0.23) wool characteristics on the britch. However, at the britch, wool from female lambs had a reduced comfort factor (P = 0.01) and a greater (P = 0.02) fiber diameter compared with wool from male lambs. Maternal Se supplementation, maternal nutritional plane, sex of the offspring, or their interactions had no effect (P > 0.13) on primary (29.10 +/- 1.40/100 um(2)) and secondary (529.84 +/- 21.57/100 um(2)) wool follicle numbers. Lambs from ASe ewes had a greater (P = 0.03) secondary:primary wool follicle ratio compared with lambs from HSe ewes (20.93 vs. 18.01 +/- 1.00). Despite similar postnatal diets, wool quality was affected by maternal Se status and the maternal nutritional plane. PMID- 21622886 TI - Genetic parameters and genotype x environment interaction for feed efficiency traits in steers fed grower and finisher diets. AB - The objective of this study was to examine the genetic parameters and genetic correlations of feed efficiency traits in steers (n = 490) fed grower or finisher diets in 2 feeding periods. A bivariate model was used to estimate phenotypic and genetic parameters using steers that received the grower and finisher diets in successive feeding periods, whereas a repeated animal model was used to estimate the permanent environmental effects. Genetic correlations between the grower-fed and finisher-fed regimens were 0.50 +/- 0.48 and 0.78 +/- 0.43 for residual feed intake (RFI) and G:F, respectively. The moderate genetic correlation between the 2 feeding regimens may indicate the presence of a genotype * environment interaction for RFI. Permanent environmental effects (expressed in percentage of phenotypic variance) were detected in the grower-fed steers for ADG (38%), DMI (30%), RFI (18%), and G:F (40%) and also in the finisher-fed steers for ADG (28%), DMI (35%), metabolic mid-weight (23%), and RFI (10%). Heritability estimates were 0.08 +/- 0.10 and 0.14 +/- 0.15 for the grower-fed steers and 0.42 +/- 0.16 and 0.40 +/- 17 for the finisher-fed steers for RFI and G:F, respectively. The dependency of the RFI on the feeding regimen may have serious implications when selecting animals in the beef industry. Because of the higher cost of grains, feed efficiency in the feedlot might be overemphasized, whereas efficiency in the cow herd and the backgrounding segments may have less emphasis. These results may also favor the retention (for subsequent breeding) of cows whose steers were efficient in the feedlot sector. Therefore, comprehensive feeding trials may be necessary to provide more insight into the mechanisms surrounding genotype * environment interaction in steers. PMID- 21622889 TI - Personalized growth and development plans for young nuclear medicine physicians. PMID- 21622890 TI - AD diagnostic guidelines updated. PMID- 21622891 TI - A president's farewell message. PMID- 21622892 TI - SNM clinical trials network validates 100th scanner. PMID- 21622893 TI - O-(2-18F-fluoroethyl)-L-tyrosine PET predicts failure of antiangiogenic treatment in patients with recurrent high-grade glioma. AB - The objective of this study was to compare MRI response assessment with metabolic O-(2-(18)F-fluoroethyl)-L-tyrosine ((18)F-FET) PET response evaluation during antiangiogenic treatment in patients with recurrent high-grade glioma (rHGG). METHODS: Eleven patients with rHGG were treated biweekly with bevacizumab irinotecan. MR images and (18)F-FET PET scans were obtained at baseline and at follow-up 8-12 wk after treatment onset. MRI treatment response was evaluated by T1/T2 volumetry according to response assessment in neurooncology (RANO) criteria. For (18)F-FET PET evaluation, an uptake reduction of more than 45% calculated with a standardized uptake value of more than 1.6 was defined as a metabolic response (receiver-operating-characteristic curve analysis). MRI and (18)F-FET PET volumetry results and response assessment were compared with each other and in relation to progression-free survival (PFS) and overall survival (OS). RESULTS: At follow-up, MR images showed partial response in 7 of 11 patients (64%), stable disease in 2 of 11 patients (18%), and tumor progression in 2 of 11 patients (18%). In contrast, (18)F-FET PET revealed 5 of 11 metabolic responders (46%) and 6 of 11 nonresponders (54%). MRI and (18)F-FET PET showed that responders survived significantly longer than did nonresponders (10.24 vs. 4.1 mo, P = 0.025, and 7.9 vs. 2.3 mo, P = 0.015, respectively). In 4 patients (36.4%), diagnosis according to RANO criteria and (18)F-FET PET was discordant. In these cases, PET was able to detect tumor progression earlier than was MRI. CONCLUSION: In rHGG patients undergoing antiangiogenic treatment, (18)F-FET PET seems to be predictive for treatment failure in that it contributes important information to response assessment based solely on MRI and RANO criteria. PMID- 21622894 TI - Nuclear medicine in the first year of life. AB - Nuclear medicine has an important role in the care of newborns and children less than 1 y old. Patients in this age group present with a spectrum of diseases different from those of older children or adults. These patients can benefit from the full range of nuclear medicine studies. In these young children, nuclear medicine studies are more likely to be used to evaluate a wide range of congenital conditions but also can be helpful for evaluating acquired conditions such as infection, cancer, and trauma. This review first will cover the general aspects of nuclear medicine practice with these patients, including the special considerations that can help achieve successful diagnostic imaging. These topics will include clinical indications, imaging technology, instrumentation, software, positioning and immobilization, sedation, local and general anesthesia, radiopharmaceutical doses, radiation risk, and dose reduction. The review then will discuss the specific nuclear medicine studies that typically are obtained in patients in this age group. With extra care and attention to the special needs of this population, nuclear medicine departments can successfully study patients less than 1 y old. PMID- 21622895 TI - In vivo long-term kinetics of radiolabeled n,n-dimethyltryptamine and tryptamine. AB - N,N-dimethyltryptamine (DMT), a strong psychodysleptic drug, has been found in higher plants, shamanic hallucinogenic beverages, and the urine of schizophrenic patients. The aim of this work was to gain better knowledge on the relationship between this drug and hallucinogenic processes by studying DMT behavior in comparison with tryptamine. METHODS: (131)I-labeled DMT and tryptamine were injected into rabbits. gamma-Camera and biodistribution studies were performed. Brain uptake, plasma clearance, and renal excretion were assessed for each indolealkylamine. RESULTS: DMT and tryptamine showed different behavior when brain uptake, residence time, and excretion were compared. Labeled DMT entered the brain 10 s after injection, crossed the blood-brain barrier, and bound to receptors; then it was partially renally excreted. It was detected in urine within 24 h after injection and remained in the brain, even after urine excretion ceased; up to 0.1% of the injected dose was detected at 7 d after injection in the olfactory bulb. In contrast, tryptamine was rapidly taken up in the brain and fully excreted 10 min after injection. CONCLUSION: To our knowledge, this is the first demonstration that exogenous DMT remains in the brain for at least 7 d after injection. Although labeled DMT and tryptamine behave as agonists for at least 5-hydroxytryptamine 2A receptor, 5-hydroxytryptamine 2C receptor, trace amine-associated receptor, and sigma-1 putative receptor targets, binding to the latter can explain the different behavior of labeled DMT and tryptamine in the brain. The persistence in the brain can be further explained on the basis that DMT and other N,N-dialkyltryptamines are transporter substrates for both the plasma membrane serotonin transporter and the vesicle monoamine transporter 2. Furthermore, storage in vesicles prevents DMT degradation by monoamine oxidase. At high concentrations, DMT is taken up by the serotonin transporter and further stored in vesicles by the vesicle monoamine transporter 2, to be released under appropriate stimuli. Moreover, the (131)I-labeling proved to be a useful tool to perform long-term in vivo studies. PMID- 21622896 TI - Imaging CXCR4 expression in human cancer xenografts: evaluation of monocyclam 64Cu-AMD3465. AB - The chemokine receptor 4 (CXCR4) is overexpressed in several cancers and metastases and as such presents an enticing target for molecular imaging of metastases and metastatic potential of the primary tumor. CXCR4-based imaging agents could also be useful for diagnosis, staging, and therapeutic monitoring. Here we evaluated a positron-emitting monocyclam analog, (64)Cu-{N-[1,4,8,11 tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine} ((64)Cu-AMD3465), in subcutaneous U87 brain tumors and U87 tumors stably expressing CXCR4 (U87-stb-CXCR4) and in colon tumors (HT-29) using dynamic and whole-body PET supported by ex vivo biodistribution studies. Both dynamic and whole-body PET/CT studies show specific accumulation of radioactivity in U87-stb CXCR4 tumors, with the percentage injected dose per gram reaching a maximum of 102.70 +/- 20.80 at 60 min and tumor-to-muscle ratios reaching a maximum of 362.56 +/- 153.51 at 90 min after injection of the radiotracer. Similar specificity was also observed in the HT-29 colon tumor model. Treatment with AMD3465 inhibited uptake of radioactivity by the tumors in both models. Our results show that (64)Cu-AMD3465 is capable of detecting lesions in a CXCR4 dependent fashion, with high target selectivity, and may offer a scaffold for the synthesis of clinically translatable agents. PMID- 21622897 TI - Proteomics identification of ITGB3 as a key regulator in reactive oxygen species induced migration and invasion of colorectal cancer cells. AB - Colorectal cancer (CRC) is the third most commonly diagnosed cancer in males and second in females worldwide. Unfortunately 40-50% of patients already have metastatic disease at presentation when prognosis is poor with a 5-year survival of <10%. Reactive oxygen species (ROS) have been proposed to play a crucial role in tumor metastasis. We now show that higher levels of ROS accumulation are found in a colorectal cancer-derived metastatic cell line (SW620) compared with a cell line (SW480) derived from the primary lesion from the same patient. In addition, ROS accumulation can affect both the migratory and invasive capacity of SW480 and SW620 cells. To explore the molecular mechanism underlying ROS-induced migration and invasion in CRC, we have compared protein expression patterns between SW480 and SW620 cells using a two-dimensional electrophoresis-based proteomics strategy. A total of 63 altered proteins were identified from tandem MS analysis. Cluster analysis revealed dysregulated expression of multiple redox regulative or ROS responsive proteins, implicating their functional roles in colorectal cancer metastasis. Molecular and pathological validation demonstrated that altered expression of PGAM1, GRB2, DJ-1, ITGB3, SOD-1, and STMN1 was closely correlated with the metastatic potential of CRC. Functional studies showed that ROS markedly up-regulated expression of ITGB3, which in turn promoted an aggressive phenotype in SW480 cells, with concomitant up-regulated expression of STMN1. In contrast, knockdown of ITGB3 expression could mitigate the migratory and invasive potential of SW620 or H(2)O(2)-treated SW480 cells, accompanied by down-regulated expression of STMN1. The function of ITGB3 was dependent on the surface expression of integrin alphavbeta3 heterodimer. Furthermore, STMN1 expression and the PI3K-Akt-mTOR pathway were found to be involved in ROS-induced and ITGB3 mediated migration and invasion of colorectal cancer cells. Taken together, these studies suggest that ITGB3 plays an important role in ROS-induced migration and invasion in CRC. PMID- 21622898 TI - The need for academic generalists. AB - Academia should be willing to shoulder some of the responsibility for the current dearth of new therapeutic drugs. Our research funding is predicated on the assumption that it will bring value to society, but our emphasis on scientific specialization hinders our ability to add value when a broader vision is required. A solution is the creation of an academy of science generalists motivated to bring together clinical and basic scientists, academia and the private sector, government legislators and industry. A small investment in academic generalists could yield benefits far beyond its modest cost. PMID- 21622899 TI - The RNR motif of B. subtilis RNase P protein interacts with both PRNA and pre tRNA to stabilize an active conformer. AB - Ribonuclease P (RNase P) catalyzes the metal-dependent 5' end maturation of precursor tRNAs (pre-tRNAs). In Bacteria, RNase P is composed of a catalytic RNA (PRNA) and a protein subunit (P protein) necessary for function in vivo. The P protein enhances pre-tRNA affinity, selectivity, and cleavage efficiency, as well as modulates the cation requirement for RNase P function. Bacterial P proteins share little sequence conservation although the protein structures are homologous. Here we combine site-directed mutagenesis, affinity measurements, and single turnover kinetics to demonstrate that two residues (R60 and R62) in the most highly conserved region of the P protein, the RNR motif (R60-R68 in Bacillus subtilis), stabilize PRNA complexes with both P protein (PRNA*P protein) and pre tRNA (PRNA*P protein*pre-tRNA). Additionally, these data indicate that the RNR motif enhances a metal-stabilized conformational change in RNase P that accompanies substrate binding and is essential for efficient catalysis. Stabilization of this conformational change contributes to both the decreased metal requirement and the enhanced substrate recognition of the RNase P holoenzyme, illuminating the role of the most highly conserved region of P protein in the RNase P reaction pathway. PMID- 21622900 TI - Polymerase ribozyme efficiency increased by G/T-rich DNA oligonucleotides. AB - The RNA world hypothesis states that the early evolution of life went through a stage where RNA served as genome and as catalyst. The replication of RNA world organisms would have been facilitated by ribozymes that catalyze RNA polymerization. To recapitulate an RNA world in the laboratory, a series of RNA polymerase ribozymes was developed previously. However, these ribozymes have a polymerization efficiency that is too low for self-replication, and the most efficient ribozymes prefer one specific template sequence. The limiting factor for polymerization efficiency is the weak sequence-independent binding to its primer/template substrate. Most of the known polymerase ribozymes bind an RNA heptanucleotide to form the P2 duplex on the ribozyme. By modifying this heptanucleotide, we were able to significantly increase polymerization efficiency. Truncations at the 3'-terminus of this heptanucleotide increased full length primer extension by 10-fold, on a specific template sequence. In contrast, polymerization on several different template sequences was improved dramatically by replacing the RNA heptanucleotide with DNA oligomers containing randomized sequences of 15 nt. The presence of G and T in the random sequences was sufficient for this effect, with an optimal composition of 60% G and 40% T. Our results indicate that these DNA sequences function by establishing many weak and nonspecific base-pairing interactions to the single-stranded portion of the template. Such low-specificity interactions could have had important functions in an RNA world. PMID- 21622902 TI - Nrf-2 regulates cyclosporine-stimulated HO-1 expression in gingiva. AB - Cyclosporine-A (CsA) stimulates heme oxygenase-1 (HO-1) expression in the gingiva, but the regulation and the role of HO-1 in gingival overgrowth are not well-understood. HO-1 is regulated by several transcription factors, such as nuclear factor-kappaB (NF-kappaB) and nuclear factor erythroid-2-related factor 2 (Nrf-2). The aim of this study was to examine the role of Nrf-2 in the regulation of CsA-stimulated HO-1 expression in human gingival fibroblasts. Nrf-2 siRNA (siNrf-2), NF-kappaB, kinase inhibitors, and sulforaphane (SFN) were used to examine the nuclear translocation of Nrf-2 and expression of HO-1 and transforming growth factor-beta1 (TGF-beta1) in cells. Treatment with siNrf-2, but not with an NF-kappaB inhibitor, reduced CsA-stimulated HO-1 mRNA expression. ERK inhibition significantly decreased CsA-stimulated Nrf-2 nuclear translocation and HO-1 mRNA expression. Pre-treatment with SFN showed that HO-1 plays a role in attenuating CsA-mediated TGF-beta1 expressions. These findings suggest that CsA stimulated HO-1 expression is mediated through the activation of ERK, and that Nrf-2 plays a protective role against CsA-induced gingival fibrosis by modulating collagen turnover-related genes. PMID- 21622901 TI - VapC6, a ribonucleolytic toxin regulates thermophilicity in the crenarchaeote Sulfolobus solfataricus. AB - The phylum Crenarchaeota includes hyperthermophilic micro-organisms subjected to dynamic thermal conditions. Previous transcriptomic studies of Sulfolobus solfataricus identified vapBC6 as a heat-shock (HS)-inducible member of the Vap toxin-antitoxin gene family. In this study, the inactivation of the vapBC6 operon by targeted gene disruption produced two recessive phenotypes related to fitness, HS sensitivity and a heat-dependent reduction in the rate of growth. In-frame vapBC6 deletion mutants were analyzed to examine the respective roles of each protein. Since vapB6 transcript abundance was elevated in the vapC6 deletion, the VapC6 toxin appears to regulate abundance of its cognate antitoxin. In contrast, vapC6 transcript abundance was reduced in the vapB6 deletion. A putative intergenic terminator may underlie these observations by coordinating vapBC6 expression. As predicted by structural modeling, recombinant VapC6 produced using chaperone cosynthesis exhibited heat-dependent ribonucleolytic activity toward S. solfataricus total RNA. This activity could be blocked by addition of preheated recombinant VapB6. In vivo transcript targets were identified by assessing the relative expression of genes that naturally respond to thermal stress in VapBC6 deficient cells. Preferential increases were observed for dppB-1 and tetR, and preferential decreases were observed for rpoD and eIF2 gamma. Specific VapC6 ribonucleolytic action could also be demonstrated in vitro toward RNAs whose expression increased in the VapBC6-deficient strain during heat shock. These findings provide a biochemical mechanism and identify cellular targets underlying VapBC6-mediated control over microbial growth and survival at temperature extremes. PMID- 21622903 TI - Role of Hsl7 in morphology and pathogenicity and its interaction with other signaling components in the plant pathogen Ustilago maydis. AB - The phytopathogenic fungus Ustilago maydis undergoes a dimorphic transition in response to mating pheromone, host, and environmental cues. On a solid medium deficient in ammonium (SLAD [0.17% yeast nitrogen base without ammonium sulfate or amino acids, 2% dextrose, 50 MUM ammonium sulfate]), U. maydis produces a filamentous colony morphology, while in liquid SLAD, the cells do not form filaments. The p21-activated protein kinases (PAKs) play a substantial role in regulating the dimorphic transition in fungi. The PAK-like Ste20 homologue Smu1 is required for a normal response to pheromone, via upregulation of pheromone expression, and virulence, and its disruption affects both processes. Our experiments suggest that Smu1 also regulates cell length and the filamentous response on solid SLAD medium. Yeast two-hybrid analysis suggested an Hsl7 homologue as a potential interacting partner of Smu1, and a unique open reading frame for such an arginine methyltransferase was detected in the U. maydis genome sequence. Hsl7 regulates cell length and the filamentous response to solid SLAD in a fashion opposite to that of Smu1, but neither overexpression nor disruption of hsl7 attenuates virulence. Simultaneous disruption of hsl7 and overexpression of smu1 lead to a hyperfilamentous response on solid SLAD. Moreover, only this double mutant strain forms filaments in liquid SLAD. The double mutant strain was also significantly reduced in virulence. A similar filamentous response in both solid and liquid SLAD was observed in strains lacking another PAK-like protein kinase involved in cytokinesis and polar growth, Cla4. Our data suggest that Hsl7 may regulate cell cycle progression, while both Smu1 and Cla4 appear to be involved in the filamentous response in U. maydis. PMID- 21622904 TI - Do red and green make brown?: perspectives on plastid acquisitions within chromalveolates. AB - The chromalveolate "supergroup" is of key interest in contemporary phycology, as it contains the overwhelming majority of extant algal species, including several phyla of key importance to oceanic net primary productivity such as diatoms, kelps, and dinoflagellates. There is also intense current interest in the exploitation of these algae for industrial purposes, such as biodiesel production. However, the evolution of the constituent species, and in particular the origin and radiation of the chloroplast genomes, remains poorly understood. In this review, we discuss current theories of the origins of the extant red alga derived chloroplast lineages in the chromalveolates and the potential ramifications of the recent discovery of large numbers of green algal genes in chromalveolate genomes. We consider that the best explanation for this is that chromalveolates historically possessed a cryptic green algal endosymbiont that was subsequently replaced by a red algal chloroplast. We consider how changing selective pressures acting on ancient chromalveolate lineages may have selectively favored the serial endosymbioses of green and red algae and whether a complex endosymbiotic history facilitated the rise of chromalveolates to their current position of ecological prominence. PMID- 21622905 TI - Effects of fluconazole on the secretome, the wall proteome, and wall integrity of the clinical fungus Candida albicans. AB - Fluconazole is a commonly used antifungal drug that inhibits Erg11, a protein responsible for 14alpha-demethylation during ergosterol synthesis. Consequently, ergosterol is depleted from cellular membranes and replaced by toxic 14alpha methylated sterols, which causes increased membrane fluidity and drug permeability. Surface-grown and planktonic cultures of Candida albicans responded similarly to fluconazole at 0.5 mg/liter, showing reduced biomass formation, severely reduced ergosterol levels, and almost complete inhibition of hyphal growth. There was no evidence of cell leakage. Mass spectrometric analysis of the secretome showed that its composition was strongly affected and included 17 fluconazole-specific secretory proteins. Relative quantification of (14)N-labeled query walls relative to a reference standard mixture of (15)N-labeled yeast and hyphal walls in combination with immunological analysis revealed considerable fluconazole-induced changes in the wall proteome as well. They were, however, similar for both surface-grown and planktonic cultures. Two major trends emerged: (i) decreased incorporation of hypha-associated wall proteins (Als3, Hwp1, and Plb5), consistent with inhibition of hyphal growth, and (ii) increased incorporation of putative wall repair-related proteins (Crh11, Pga4, Phr1, Phr2, Pir1, and Sap9). As exposure to the wall-perturbing drug Congo red led to a similar response, these observations suggested that fluconazole affects the wall. In keeping with this, the resistance of fluconazole-treated cells to wall perturbing compounds decreased. We propose that fluconazole affects the integrity of both the cellular membranes and the fungal wall and discuss its potential consequences for antifungal therapy. We also present candidate proteins from the secretome for clinical marker development. PMID- 21622907 TI - Obesity and the workplace. PMID- 21622909 TI - The mill reek in 1754. PMID- 21622910 TI - Estelle Sylvia Pankhurst. The Chainmaker 1907. PMID- 21622914 TI - How I use it: the Internet. PMID- 21622911 TI - Surveillance for work-related audiological disease in the UK: 1998-2006. AB - BACKGROUND: Noise-induced hearing loss (NIHL) from prolonged occupational exposure to noise continues to rank among the top worldwide work-related ill health problems. AIMS: To provide an overview of incident cases based on work related audiological ill-health data collected over a 9-year period from occupational physicians (OPs), audiological physicians (APs), general practitioners and otorhinolaryngologists. METHODS: Analysis of numerator data reported by physicians to surveillance schemes within The Health and Occupation Reporting network (THOR). The actual cases were multiplied by the sampling ratio to provide estimated numerator numbers, followed by calculation of incidence rates using denominator data derived from the Labour Force Survey and from surveys of participating OPs. RESULTS: Two thousand five hundred and eighty-two estimated cases (2584 estimated diagnoses) were received from OPs (Occupational Physicians Reporting Activity [OPRA]), and 2192 estimated cases (3208 estimated diagnoses) of work-related audiological ill-health were received from consultant APs [Occupational Surveillance Scheme for Audiological physicians (OSSA)] from 1998 to 2006. Cases where the causal agent was noise exposure (NIHL and tinnitus) made up of 95 and 97% of all cases reported to OPRA and OSSA, respectively. The annual average incidence rate for noise-induced audiological disorders was 7.9 [95% confidence interval (CI) 4.6-11.1] per 100 000 persons employed (OPRA) and 0.8 (95% CI 0.7-1.0) per 100 000 persons employed (OSSA). Workers with the highest incidence were older males employed in public administration and defence and the manufacture of metals. CONCLUSIONS: THOR data show that diagnoses related to work-related noise exposure (NIHL/tinnitus), as reported to THOR, remain important health problems, despite preventive measures being in place. PMID- 21622918 TI - Management of minor head injury in children: searching for the answer. PMID- 21622917 TI - Positional information resolves structural variations and uncovers an evolutionarily divergent genetic locus in accessions of Arabidopsis thaliana. AB - Genome sequencing of closely related individuals has yielded valuable insights that link genome evolution to phenotypic variations. However, advancement in sequencing technology has also led to an escalation in the number of poor quality drafted genomes assembled based on reference genomes that can have highly divergent or haplotypic regions. The self-fertilizing nature of Arabidopsis thaliana poses an advantage to sequencing projects because its genome is mostly homozygous. To determine the accuracy of an Arabidopsis drafted genome in less conserved regions, we performed a resequencing experiment on a ~371-kb genomic interval in the Landsberg erecta (Ler-0) accession. We identified novel structural variations (SVs) between Ler-0 and the reference accession Col-0 using a long-range polymerase chain reaction approach to generate an Illumina data set that has positional information, that is, a data set with reads that map to a known location. Positional information is important for accurate genome assembly and the resolution of SVs particularly in highly duplicated or repetitive regions. Sixty-one regions with misassembly signatures were identified from the Ler-0 draft, suggesting the presence of novel SVs that are not represented in the draft sequence. Sixty of those were resolved by iterative mapping using our data set. Fifteen large indels (>100 bp) identified from this study were found to be located either within protein-coding regions or upstream regulatory regions, suggesting the formation of novel alleles or altered regulation of existing genes in Ler-0. We propose future genome-sequencing experiments to follow a clone-based approach that incorporates positional information to ultimately reveal haplotype specific differences between accessions. PMID- 21622919 TI - ECG interference from the iPhone. PMID- 21622920 TI - Nutmeg liver. PMID- 21622921 TI - Acute disseminated encephalomyelitis: a diagnosis to consider. PMID- 21622922 TI - Trauma toys: an adjunct to taking a history. PMID- 21622923 TI - Hypocalcemia following treatment for hyperthyroidism. PMID- 21622924 TI - Commentary on: hypocalcemia following treatment for hyperthyroidism. PMID- 21622925 TI - Commentary on: hypocalcemia following treatment for hyperthyroidism. PMID- 21622926 TI - Inspiring minds. Marilyn Huestis. PMID- 21622927 TI - What is your guess? A four-day-old boy with an abnormal metabolic screen. PMID- 21622932 TI - Economic burden of injuries in South Korea. AB - BACKGROUND STUDIES: on the cost of injury are useful in setting research and policy priorities and it is valuable to observe differences in the economic burden of injuries across countries. OBJECTIVE: To estimate the treated prevalence rate, economic burden and gender- and age-specific costs of injuries in Korea in 2006. DESIGN: Annual direct healthcare costs associated with injuries were estimated from the National Health Insurance, Medical Aid and Automobile Insurance databases. Annual direct non-health costs were estimated for transport and caregiver's costs. Indirect costs were estimated for premature death, absence from work and disability. Costs were adjusted to 2006 levels using the healthcare component of the Consumer Price Index. MAIN OUTCOME MEASURES: Prevalence-based direct costs, incidence-based indirect costs and total costs for injuries, stratified by gender, age group and type of injury. RESULTS: The treated prevalence rate of injury in 2006 of the Korean population was 26.5 per 100, resulting in an annual economic burden of $39837 million ($4703 million in direct and $35134 million in indirect costs). The cost of medical treatment associated with injuries accounted for 9.5% of the total health expenditure in Korea. The cost of premature death was the largest contributor to the total and automobile related injuries accounted for 30.3% of total costs. CONCLUSIONS: The estimates were considerably understated because they did not include losses in household production and quality of life. Nevertheless, the size and main components of the injury burden were identified; this information should aid decision-making about research priorities and improve monitoring of the effects of policy initiatives. PMID- 21622933 TI - User perspectives on the usability of a regional health information exchange. AB - OBJECTIVE: We assessed the usability of a health information exchange (HIE) in a densely populated metropolitan region. This grant-funded HIE had been deployed rapidly to address the imminent needs of the patient population and the need to draw wider participation from regional entities. DESIGN: We conducted a cross sectional survey of individuals given access to the HIE at participating organizations and examined some of the usability and usage factors related to the technology acceptance model. MEASUREMENTS: We probed user perceptions using the Questionnaire for User Interaction Satisfaction, an author-generated Trust scale, and user characteristic questions (eg, age, weekly system usage time). RESULTS: Overall, users viewed the system favorably (ratings for all usability items were greater than neutral (one-sample Wilcoxon test, p<0.0014, Bonferroni-corrected for 35 tests). System usage was regressed on usability, trust, and demographic and user characteristic factors. Three usability factors were positively predictive of system usage: overall reactions (p<0 0.01), learning (p<0.05), and system functionality (p<0.01). Although trust is an important component in collaborative relationships, we did not find that user trust of other participating healthcare entities was significantly predictive of usage. An analysis of respondents' comments revealed ways to improve the HIE. CONCLUSION: We used a rapid deployment model to develop an HIE and found that perceptions of system usability were positive. We also found that system usage was predicted well by some aspects of usability. Results from this study suggest that a rapid development approach may serve as a viable model for developing usable HIEs serving communities with limited resources. PMID- 21622934 TI - The Yale cTAKES extensions for document classification: architecture and application. AB - BACKGROUND: Open-source clinical natural-language-processing (NLP) systems have lowered the barrier to the development of effective clinical document classification systems. Clinical natural-language-processing systems annotate the syntax and semantics of clinical text; however, feature extraction and representation for document classification pose technical challenges. METHODS: The authors developed extensions to the clinical Text Analysis and Knowledge Extraction System (cTAKES) that simplify feature extraction, experimentation with various feature representations, and the development of both rule and machine learning based document classifiers. The authors describe and evaluate their system, the Yale cTAKES Extensions (YTEX), on the classification of radiology reports that contain findings suggestive of hepatic decompensation. RESULTS AND DISCUSSION: The F(1)-Score of the system for the retrieval of abdominal radiology reports was 96%, and was 79%, 91%, and 95% for the presence of liver masses, ascites, and varices, respectively. The authors released YTEX as open source, available at http://code.google.com/p/ytex. PMID- 21622935 TI - Replacement of the myotonic dystrophy type 1 CTG repeat with 'non-CTG repeat' insertions in specific tissues. AB - BACKGROUND: Recently, curious mutations have been reported to occur within the (CTG)n repeat tract of the myotonic dystrophy type 1 (DM1) locus. For example, the repeat, long presumed to be a pure repeat sequence, has now been revealed to often contain interruption motifs in a proportion of cases with expansions. Similarly, a few de novo somatic CTG expansions have been reported to arise from non-expanded DM1 alleles with 5-37 units, thought to be genetically stable. AIMS AND METHODS: This study has characterised a novel mutation configuration at the DM1 CTG repeat that arose as somatic mosaicism in a juvenile onset DM1 patient with a non-expanded allele of (CTG)12 and tissue specific expansions ranging from (CTG)1100 to 6000. RESULTS: The mutation configuration replaced the CTG tract with a non-CTG repeat insertion of 43 or 60 nucleotides, precisely placed in the position of the CTG tract with proper flanking sequences. The inserts appeared to arise from a longer human sequence on chromosome 4q12, and may have arisen through DNA structure mediated somatic inter-gene recombination or replication/repair template switching errors. De novo insertions were detected in cerebral cortex and skeletal muscle, but not in heart or liver. Repeat tracts with -1 or -2 CTG units were also detected in cerebellum, which may have arisen by contractions of the short (CTG)12 allele. CONCLUSION: This non-CTG configuration expands current understanding of the sequence variations that can arise at this hypermutable site. PMID- 21622936 TI - Serum and CSF N-acetyl aspartate levels differ in multiple sclerosis and neuromyelitis optica. AB - BACKGROUND: The identification of biomarkers able to improve the differential diagnosis between multiple sclerosis (MS) and neuromyelitis optica (NMO) is challenging because of a different prognosis and response to treatment. Growing evidence indicates that brain and CSF N-acetyl aspartate (NAA) concentration is a useful marker for characterising different phases of axonal pathology in demyelinating diseases, and preliminary studies suggest that increased serum NAA levels may be a telltale sign of acute neuronal damage or defective NAA metabolism in oligodendrocytes. OBJECTIVE: To evaluate whether serum and CSF NAA concentration differs in patients with MS and NMO. DESIGN: Observational, multicentre, prospective, cross sectional study. METHODS: Serum samples were collected from 48 relapsing-remitting MS, 32 NMO and 76 age matched healthy controls. Coeval CSF samples were available for all MS and for 8/32 NMO patients. NAA was measured in serum and CSF by liquid chromatography-mass spectrometry. RESULTS: MS patients showed higher serum and CSF NAA levels than NMO patients, and higher serum NAA levels than healthy controls (p<0.001). High serum NAA values, exceeding the 95th percentile of serum NAA values in healthy controls, were found in 100% of patients with MS and in no patient with NMO. No differences in serum NAA levels were found between NMO and healthy controls. In MS, serum and CSF NAA levels correlated with disability score. CONCLUSIONS: Determination of serum and CSF NAA levels may represent a suitable tool in the diagnostic laboratory workup to differentiate MS and NMO. PMID- 21622937 TI - Population based epidemiology of amyotrophic lateral sclerosis using capture recapture methodology. AB - BACKGROUND: Variation in the incidence rate in epidemiological studies on amyotrophic lateral sclerosis (ALS) may be due to a small population size and under ascertainment of patients. The previously reported incidence decline in the elderly and a decrease in the male:female ratio in postmenopausal age groups have yet to be confirmed. METHODS: ALS epidemiology in a large population based register in The Netherlands was studied between 1 January 2006 and 31 December 2009, and applied capture-recapture methodology in separate age and gender groups to adjust for the number of unobserved patients. RESULTS: 1217 incident patients were observed, and a capture-recapture incidence of 2.77 per 100 000 person-years (95% CI 2.63 to 2.91). Prevalence on 31 December 2008 was 10.32 per 100 000 individuals (95% CI 9.78 to 10.86). The incident cohort had a higher median age at onset (63.0 vs 58.1 years) and more bulbar onset patients (30.0% vs 19.1%) compared with the prevalent cohort. Incidence and prevalence peaked in the 70-74 year age group followed by a rapid decline in older age. The male:female ratio in the premenopausal age group (1.91, 95% CI 1.32 to 2.79) was not significantly higher than that in the postmenopausal age group (1.50, 95% CI 1.34 to 1.67). CONCLUSION: The marked difference in patient characteristics between incident and prevalent cohorts underscores the importance of including incident patients when studying susceptibility or disease modifying factors in ALS. The incidence decline in the elderly may suggest that ALS is not merely the result of ageing. Absence of a significant postmenopausal drop in the male:female ratio suggests that the protective role of female sex hormones in ALS is limited. PMID- 21622938 TI - Stage classification and prognosis: an intersection of medicine, quantum physics and religion? AB - Estimating prognosis is an important part of caring for patients with cancer. However, predicting prognosis is complicated and depends on many factors. Simply amassing more data alone is not the answer; we have to learn to intellectually manage the inherent complexity and uncertainty if we are to make progress. PMID- 21622939 TI - A qualitative investigation of selecting surrogate decision-makers. AB - BACKGROUND: Empirical studies of surrogate decision-making tend to assume that surrogates should make only a 'substituted judgement'--that is, judge what the patient would want if they were mentally competent. OBJECTIVES: To explore what people want in a surrogate decision-maker whom they themselves select and to test the assumption that people want their chosen surrogate to make only a substituted judgement. METHODS: 30 undergraduate students were recruited. They were presented with a hypothetical scenario about their expected loss of mental capacity in the future and asked to answer some questions about their choice of surrogate. These data were analysed qualitatively using thematic content analysis. RESULTS: Most respondents talked about choosing someone who was caring and competent in certain ways, giving interesting evidence for their judgements. Surprisingly few highlighted how well they thought their chosen surrogate knew their preferences and would be able to make a substituted judgement. Moreover, few specified that their chosen surrogate had similar attitudes and values to their own and so would make a similar decision to theirs in the circumstances presented. Some respondents also referred to the social role of their chosen surrogate or the social dynamics of their situation which influenced their choices, as well as to ideas of reciprocity and characteristics of honesty and loyalty. CONCLUSION: In the event that they lose mental capacity, many people will not select a surrogate to decide about medical treatments on their behalf solely on the basis that they expect their surrogate to make a substituted judgement. PMID- 21622940 TI - Single-nucleotide polymorphisms in DNA repair genes and association with breast cancer risk in the web study. AB - Base excision repair (BER) and nucleotide excision repair (NER) pathways repair damaged DNA, and polymorphisms in these genes might affect breast cancer susceptibility. We evaluated associations between seven single-nucleotide polymorphisms in four DNA repair genes (ERCC4 rs1799801, XPC rs2227998, rs2228001, rs2228000, OGG1 rs1052133 and XRCC1 rs25487 and rs25486) and breast cancer risk, examining modification by smoking and alcohol consumption, using data from the Western New York Exposures and Breast Cancer Study. Women aged 35 79 years with incident breast cancer (n = 1170) and age- and race-matched controls (n = 2115) were enrolled. Genotyping was performed using matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Unconditional logistic regression was used to estimate odds ratios (OR) and 95% confidence intervals (CIs). No significant associations were observed in premenopausal women. Among postmenopausal women, rs25487 and rs25486 (OR = 1.24; 95% CI 1.01 1.51 and OR = 1.23; 95% CI 1.01-1.49, respectively, for combined heterozygous and homozygous variant compared with reference) were associated with increased risk of breast cancer. Postmenopausal women carrying the variant allele of the synonymous XPC polymorphism (rs2227998) were also at borderline significantly increased risk (OR = 1.24; 95% CI 1.01-1.52, heterozygous variant compared with reference; OR = 1.22; 95% CI 1.01-1.48, for combined heterozygous and homozygous variant compared with reference). There was no evidence of genotype-smoking and genotype-alcohol consumption interactions for pre- and postmenopausal women. These results indicate that some of the variants in BER and NER genes may influence risk of postmenopausal breast cancer. PMID- 21622941 TI - Site-specific in vivo mutagenicity in the kidney of gpt delta rats given a carcinogenic dose of ochratoxin A. AB - Ochratoxin A (OTA) can induce renal tumors that originate from the S3 segment of the proximal tubules in rodents, but the results of conventional mutagenicity tests have caused controversy regarding the role of genotoxic mechanisms in the carcinogenesis. Human exposure to OTA from various foods is unavoidable. Therefore, an understanding of OTA-induced renal carcinogenesis is necessary for accurate estimates of the human risk hazard. In the present study, a 13-week exposure of gpt delta rats to OTA at a carcinogenic dose induced karyomegaly and apoptosis at the outer stripe of the outer medulla (OM) of the kidney but failed to affect the reporter gene mutations in DNA extracted from whole kidneys. This site specificity resulting from the kinetics of specific transporters might be responsible for the negative outcome of in vivo mutagenicity. The kidney was then macroscopically divided, based on anatomical characteristics, into the cortex, the OM, and the inner medulla, each of which was histopathologically confirmed. Spi- mutant frequencies (MFs) but not gpt MFs in the OM after a 4-week exposure to OTA were significantly higher than in controls despite the absence of cortical changes. There were also no changes in 8-hydroxydeoxyguanosine levels in kidney DNA. These results strongly suggest the involvement of a genotoxic mechanism, with the exception of oxidative DNA damage in OTA-induced renal carcinogenesis. In addition, the reporter gene mutation assay using DNA from target sites could be a more powerful tool to investigate in vivo genotoxicities. PMID- 21622942 TI - Characterization of novel ligands of ERalpha, Erbeta, and PPARgamma: the case of halogenated bisphenol A and their conjugated metabolites. AB - The capability of the flame retardants tetrabromobisphenol A (TBBPA) and tetrachlorobisphenol A (TCBPA) to activate peroxysome proliferator-activated receptors (PPARs) alpha, beta, and gamma and estrogen receptors (ERs) alpha and beta has been recently investigated, but the activity of their biotransformation products and of their lower molecular weight analogues formed in the environment remains unexplored. The aim of this study was to investigate the relationship between the degree of halogenation of BPA analogues and their affinity and activity towards human PPARgamma and ERs and to characterize active metabolites of major marketed halogenated bisphenols. The biological activity of all compounds was studied using reporter cell lines expressing these nuclear receptors (NRs). We used NR-based affinity columns to rapidly evaluate the binding affinity of halogenated bisphenols for PPARgamma and ERs and to trap active metabolites of TBBPA and TCBPA formed in HepG2 cells. The agonistic potential of BPA analogs highly depends on their halogenation degree: the bulkier halogenated BPA analogs, the greater their capability to activate PPARgamma. In addition, PPARgamma-based affinity column, HGELN-PPARgamma reporter cell line and crystallographic analysis clearly demonstrate that the sulfation pathway, usually considered as a detoxification process, leads for TBBPA and TCBPA, to the formation of sulfate conjugates which possess a residual PPARgamma-binding activity. Our results highlight the effectiveness NR-based affinity columns to trap and characterize biologically active compounds from complex matrices. Polyhalogenated bisphenols, but also some of their metabolites, are potential disrupters of PPARgamma activity. PMID- 21622943 TI - Citrinin-generated reactive oxygen species cause cell cycle arrest leading to apoptosis via the intrinsic mitochondrial pathway in mouse skin. AB - The mycotoxin, citrinin (CTN), is a contaminant of various food and feed materials. Several in vivo and in vitro studies have demonstrated that CTN has broad toxicity spectra; however, dermal toxicity is not known. In the present investigation, dermal exposure to CTN was undertaken to study oxidative stress, DNA damage, cell cycle arrest, and apoptosis in mouse skin. A single topical application of CTN caused significant change in oxidative stress markers, such as lipid peroxidation, protein carbonyl content, glutathione (GSH) content, and antioxidant enzymes in a dose-dependent (25-100 MUg/mouse) and time-dependent (12 72 h) manner. Single topical application of CTN (50 MUg/mouse) for 12-72 h caused significant enhancement in (1) reactive oxygen species (ROS); (2) cell cycle arrest at the G0/G1 phase (30-71%) and G2/M phase (56-65%) along with the induction of apoptosis (3.6-27%); (3) expression of p53, p21/waf1; (4) Bax/Bcl2 ratio and cytochome c release; and (5) activities of caspase 9 (22-46%) and 3 (42 54%) as well as increased poly(ADP-ribose) polymerase cleavage. It was also observed that pretreatment with bio-antioxidants viz butylated hydroxyanisole (55 MUmol/100 MUl), quercetin (10 MUmol/100 MUl), or alpha-tocopherol (40 MUmol/100 MUl) resulted in decreases of ROS generation, arrest in the G0/G1 phase of the cell cycle, and apoptosis. These data confirm the involvement of ROS in apoptosis and suggest that these bio-antioxidants may be useful in the prevention of CTN induced dermal toxicity. PMID- 21622944 TI - Physiologically based pharmacokinetic modeling of fetal and neonatal manganese exposure in humans: describing manganese homeostasis during development. AB - Concerns for potential vulnerability to manganese (Mn) neurotoxicity during fetal and neonatal development have been raised due to increased needs for Mn for normal growth, different sources of exposure to Mn, and pharmacokinetic differences between the young and adults. A physiologically based pharmacokinetic (PBPK) model for Mn during human gestation and lactation was developed to predict Mn in fetal and neonatal brain using a parallelogram approach based upon extrapolation across life stages in rats and cross-species extrapolation to humans. Based on the rodent modeling, key physiological processes controlling Mn kinetics during gestation and lactation were incorporated, including alterations in Mn uptake, excretion, tissue-specific distributions, and placental and lactational transfer of Mn. Parameters for Mn kinetics were estimated based on human Mn data for milk, placenta, and fetal/neonatal tissues, along with allometric scaling from the human adult model. The model was evaluated by comparison with published Mn levels in cord blood, milk, and infant blood. Maternal Mn homeostasis during pregnancy and lactation, placenta and milk Mn, and fetal/neonatal tissue Mn were simulated for normal dietary intake and with inhalation exposure to environmental Mn. Model predictions indicate similar or lower internal exposures to Mn in the brains of fetus/neonate compared with the adult at or above typical environmental air Mn concentrations. This PBPK approach can assess expected Mn tissue concentration during early life and compares contributions of different Mn sources, such as breast or cow milk, formula, food, drinking water, and inhalation, with tissue concentration. PMID- 21622946 TI - The instantaneous center of rotation of the mandible in nonhuman primates. AB - Kinematic analyses of mandibular movement in humans demonstrate that the mandibular instantaneous center of rotation (ICoR) is commonly located near the level of the occlusal plane and varies in its position during a chewing sequence. Few data are available regarding the location of the ICoR in nonhuman primates and it remains unclear how the position of the ICoR varies in association with mastication and/or gape behaviors. ICoR was quantified throughout the gape cycle in five species of nonhuman primates (Macaca mulatta, Cebus apella, Chlorocebus aethiops, Eulemur fulvus, and Varecia variegata). The ICoR is commonly located below the mandibular condyle close to the occlusal plane and varies considerably both superoinferiorly and anteroposteriorly through the gape cycle. The path of the ICoR, and by inference condylar movement, in Macaca and Chlorocebus differs from humans whereas movement in Cebus resembles that of humans. Similarities between humans and Cebus in articular eminence and occlusal morphology may explain these resemblances. Food material properties had little influence on ICoR movement parameters. PMID- 21622945 TI - Organotins are potent activators of PPARgamma and adipocyte differentiation in bone marrow multipotent mesenchymal stromal cells. AB - Adipocyte differentiation in bone marrow is potentially deleterious to both bone integrity and lymphopoiesis. Here, we examine the hypothesis that organotins, common environmental contaminants that are dual ligands for peroxisome proliferator-activated receptor (PPAR) gamma and its heterodimerization partner retinoid X receptor (RXR), are potent activators of bone marrow adipogenesis. A C57Bl/6-derived bone marrow multipotent mesenchymal stromal cell (MSC) line, BMS2, was treated with rosiglitazone, a PPARgamma agonist, bexarotene, an RXR agonist, or a series of organotins. Rosiglitazone and bexarotene potently activated adipocyte differentiation; however, bexarotene had a maximal efficacy of only 20% of that induced by rosiglitazone. Organotins (tributyltin [TBT], triphenyltin, and dibutyltin) also stimulated adipocyte differentiation (EC50 of 10-20 nM) but with submaximal, structure-dependent efficacy. In coexposures, both bexarotene and TBT enhanced rosiglitazone-induced adipogenesis. To investigate the contribution of PPARgamma to TBT-induced adipogenesis, we examined expression of PPARgamma2, as well as its transcriptional target FABP4. TBT-induced PPARgamma2 and FABP4 protein expression with an efficacy intermediate between rosiglitazone and bexarotene, similar to lipid accumulation. A PPARgamma antagonist and PPARgamma-specific small hairpin RNA suppressed TBT-induced differentiation, although to a lesser extent than rosiglitazone-induced differentiation, suggesting that TBT may engage alternate pathways. TBT and bexarotene, but not rosiglitazone, also induced the expression of TGM2 (an RXR target) and ABCA1 (a liver X receptor target). The results show that an environmental contaminant, acting with the same potency as a therapeutic drug, induces PPARgamma-dependent adipocyte differentiation in bone marrow MSCs. Activation of multiple nuclear receptor pathways by organotins may have significant implications for bone physiology. PMID- 21622947 TI - Effect of flexural and torsional wing flexibility on lift generation in hoverfly flight. AB - The effect of wing flexibility in hoverflies was investigated using an at-scale mechanical model. Unlike dynamically-scaled models, an at-scale model can include all phenomena related to motion and deformation of the wing during flapping. For this purpose, an at-scale polymer wing mimicking a hoverfly was fabricated using a custom micromolding process. The wing has venation and corrugation profiles which mimic those of a hoverfly wing and the measured flexural stiffness of the artificial wing is comparable to that of the natural wing. To emulate the torsional flexibility at the wing-body joint, a discrete flexure hinge was created. A range of flexure stiffnesses was chosen to match the torsional stiffness of pronation and supination in a hoverfly wing. The polymer wing was compared with a rigid, flat, carbon-fiber wing using a flapping mechanism driven by a piezoelectric actuator. Both wings exhibited passive rotation around the wing hinge; however, these rotations were reduced in the case of the compliant polymer wing due to chordwise deformations during flapping which caused a reduced effective angle of attack. Maximum lift was achieved when the stiffness of the hinge was similar to that of a hoverfly in both wing cases and the magnitude of measured lift is sufficient for hovering; the maximum lift achieved by the single polymer and carbon-fiber wings was 5.9 * 10(2)( )MUN and 6.9 * 10(2)( )MUN, respectively. These results suggest that hoverflies could exploit intrinsic compliances to generate desired motions of the wing and that, for the same flapping motions, a rigid wing could be more suitable for producing large lift. PMID- 21622948 TI - Changes in the prevalence of cognitive impairment among older Americans, 1993 2004: overall trends and differences by race/ethnicity. AB - The authors used data from 6 waves of the Health and Retirement Study to evaluate changes in the prevalence of cognitive impairment among adults 70 years of age or older from 1993 to 2004. Having sampling weights for each wave enabled the authors to create merged waves that represented cross-sections of the community dwelling older population for that year. Logistic regression analyses with year as the predictor were used to estimate trends and determine the contribution of sociodemographic and health status variables to decreasing trends in the prevalence of cognitive impairment over time (score <=8 on a modified Telephone Interview Cognitive Screen). Results showed an annual decline in the prevalence of cognitive impairment of 3.4% after adjustment for age, gender, and prior test exposure (odds ratio (OR) = 0.966, 95% confidence interval (CI): 0.941, 0.992). The addition of socioeconomic variables to the model attenuated the trend by 72.1%. The annual percentage of decline in impairment was larger for blacks (OR = 0.943, 95% CI: 0.914, 0.973) and Hispanics (OR = 0.954, 95% CI: 0.912, 0.997) than for whites (OR = 0.971, 95% CI: 0.936, 1.006), although the differences were not statistically significant. Linear probability models used in secondary analyses showed larger percentage-point declines for blacks and Hispanics. Improvements in educational level contributed to declines in cognitive impairment among older adults-particularly blacks and Hispanics-in the United States. PMID- 21622949 TI - Association between urban life-years and cardiometabolic risk: the Indian migration study. AB - Urban living is associated with an increase in cardiometabolic risks, but the speed at which these risks are accrued over time is unknown. Using a cross sectional sibling-pair design, the authors surveyed migrant factory workers and their spouses from 4 cities in India together with their rural-dwelling siblings and examined the associations between urban life-years and cardiometabolic risk factors. Data on 4,221 participants (39% women; mean age = 41 years) were available (2005-2007). In regression models, a 2-slope pattern for body fat (with a marked shift at 10 years) was found, whereas a common slope could be accepted for other risk factors. In men, the regression coefficients (per decade of urban life) were 2.5% in the first decade and 0.1% thereafter for body fat; 1.4 mm Hg for systolic blood pressure; and 7% for fasting insulin. Age, gender, marital status, household structure, and occupation did not influence the patterns appreciably; however, stronger gradients for adiposity were noted in migrants from lower socioeconomic positions. The findings suggest that body fat increases rapidly when one first moves to an urban environment, whereas other cardiometabolic risk factors evolve gradually. Public health interventions focused on the control of obesity in newer migrants to urban areas, particularly those from lower socioeconomic positions, may be beneficial. PMID- 21622950 TI - Maternal concentrations of insulin-like growth factor I and insulin-like growth factor binding protein 1 during pregnancy and birth weight of offspring. AB - Maternal concentrations of insulin-like growth factor I (IGF-I) and insulin-like growth factor binding protein 1 (IGFBP-1) may influence fetal growth. Offspring birth weight related to maternal IGF-I and IGFBP-1 measured in pregnancy was studied in 368 randomly selected women without preeclampsia who delivered a singleton liveborn child in Norway between 1992 and 1994. Maternal IGF-I concentrations were not consistently associated with birth weight, but a 1 standard deviation stronger increase in IGF-I from the first to second trimester was associated with an 82-g (95% confidence interval (CI): 11, 153) higher birth weight. IGFBP-1 concentrations were inversely associated with birth weight: Birth weight was 71 g (95% CI: 14, 128) lower per 1-standard deviation higher IGFBP-1 in the second trimester, and an increase in IGFBP-1 from the first (below median) to second (above median) trimester was associated with a 342-g (95% CI: 124, 560) lower birth weight, compared with having low IGFBP-1 (below median) in both trimesters. Conversely, low IGFBP-1 in both trimesters was associated with a 200 350-g higher birth weight compared with other combinations of IGFBP-1. In conclusion, persistently low IGFBP-1 in pregnancy is associated with relatively higher birth weight. Maternal insulin resistance may provide a link between IGFBP 1 and offspring birth weight. PMID- 21622951 TI - Functional conservation in human and Drosophila of Metazoan ADAR2 involved in RNA editing: loss of ADAR1 in insects. AB - Flies with mutations in the single Drosophila Adar gene encoding an RNA editing enzyme involved in editing 4% of all transcripts have severe locomotion defects and develop age-dependent neurodegeneration. Vertebrates have two ADAR-editing enzymes that are catalytically active; ADAR1 and ADAR2. We show that human ADAR2 rescues Drosophila Adar mutant phenotypes. Neither the short nuclear ADAR1p110 isoform nor the longer interferon-inducible cytoplasmic ADAR1p150 isoform rescue walking defects efficiently, nor do they correctly edit specific sites in Drosophila transcripts. Surprisingly, human ADAR1p110 does suppress age-dependent neurodegeneration in Drosophila Adar mutants whereas ADAR1p150 does not. The single Drosophila Adar gene was previously assumed to represent an evolutionary ancestor of the multiple vertebrate ADARs. The strong functional similarity of human ADAR2 and Drosophila Adar suggests rather that these are true orthologs. By a combination of direct cloning and searching new invertebrate genome sequences we show that distinct ADAR1 and ADAR2 genes were present very early in the Metazoan lineage, both occurring before the split between the Bilateria and Cnidarians. The ADAR1 gene has been lost several times, including during the evolution of insects and crustacea. These data complement our rescue results, supporting the idea that ADAR1 and ADAR2 have evolved highly conserved, distinct functions. PMID- 21622952 TI - ARISTO: ontological classification of small molecules by electron ionization-mass spectrometry. AB - Gas chromatography-mass spectrometry (GC-MS) acquisitions routinely yield hundreds to thousands of Electron Ionization (EI) mass spectra. The chemical identification of these spectra typically involves a search protocol that seeks an exact match to a reference spectrum. Reference spectra are found in comprehensive libraries of small molecule EI spectra curated by commercial and public entities. We developed ARISTO (Automatic Reduction of Ion Spectra To Ontology), a webtool, which provides information regarding the general chemical nature of the compound underlying an input EI mass spectrum. Importantly, ARISTO can provide such annotation without necessitating an exact match to a specific compound. ARISTO provides assignments to a subset of the ChEBI (Chemical Entities of Biological Interest) dictionary, an ontology, which aims to cover biologically relevant small molecules. Our system takes as input a mass spectrum represented as a series of mass and intensity pairs; the system returns a graphical representation of the supported ontology as well as a detailed table of suggested annotations along with their associated statistical evidence. ARISTO is accessible at this URL: http://www.ionspectra.org/aristo. The system is free, open to all and does not require registration of any sort. PMID- 21622953 TI - ICSNPathway: identify candidate causal SNPs and pathways from genome-wide association study by one analytical framework. AB - Genome-wide association study (GWAS) is widely utilized to identify genes involved in human complex disease or some other trait. One key challenge for GWAS data interpretation is to identify causal SNPs and provide profound evidence on how they affect the trait. Currently, researches are focusing on identification of candidate causal variants from the most significant SNPs of GWAS, while there is lack of support on biological mechanisms as represented by pathways. Although pathway-based analysis (PBA) has been designed to identify disease-related pathways by analyzing the full list of SNPs from GWAS, it does not emphasize on interpreting causal SNPs. To our knowledge, so far there is no web server available to solve the challenge for GWAS data interpretation within one analytical framework. ICSNPathway is developed to identify candidate causal SNPs and their corresponding candidate causal pathways from GWAS by integrating linkage disequilibrium (LD) analysis, functional SNP annotation and PBA. ICSNPathway provides a feasible solution to bridge the gap between GWAS and disease mechanism study by generating hypothesis of SNP -> gene -> pathway(s). The ICSNPathway server is freely available at http://icsnpathway.psych.ac.cn/. PMID- 21622954 TI - Swimming into peptidomimetic chemical space using pepMMsMIMIC. AB - pepMMsMIMIC is a novel web-oriented peptidomimetic compound virtual screening tool based on a multi-conformers three-dimensional (3D)-similarity search strategy. Key to the development of pepMMsMIMIC has been the creation of a library of 17 million conformers calculated from 3.9 million commercially available chemicals collected in the MMsINC(r) database. Using as input the 3D structure of a peptide bound to a protein, pepMMsMIMIC suggests which chemical structures are able to mimic the protein-protein recognition of this natural peptide using both pharmacophore and shape similarity techniques. We hope that the accessibility of pepMMsMIMIC (freely available at http://mms.dsfarm.unipd.it/pepMMsMIMIC) will encourage medicinal chemists to de peptidize protein-protein recognition processes of biological interest, thus increasing the potential of in silico peptidomimetic compound screening of known small molecules to expedite drug development. PMID- 21622955 TI - Nucleosomes protect DNA from DNA methylation in vivo and in vitro. AB - Positioned nucleosomes limit the access of proteins to DNA. However, the impact of nucleosomes on DNA methylation in vitro and in vivo is poorly understood. Here, we performed a detailed analysis of nucleosome binding and nucleosomal DNA methylation by the de novo methyltransferases. We show that compared to linker DNA, nucleosomal DNA is largely devoid of CpG methylation. ATP-dependent chromatin remodelling frees nucleosomal CpG dinucleotides and renders the remodelled nucleosome a 2-fold better substrate for Dnmt3a methyltransferase compared to free DNA. These results reflect the situation in vivo, as quantification of nucleosomal DNA methylation levels in HeLa cells shows a 2-fold decrease of nucleosomal DNA methylation levels compared to linker DNA. Our findings suggest that nucleosomal positions are stably maintained in vivo and nucleosomal occupancy is a major determinant of global DNA methylation patterns in vivo. PMID- 21622956 TI - The biological and structural characterization of Mycobacterium tuberculosis UvrA provides novel insights into its mechanism of action. AB - Mycobacterium tuberculosis is an extremely well adapted intracellular human pathogen that is exposed to multiple DNA damaging chemical assaults originating from the host defence mechanisms. As a consequence, this bacterium is thought to possess highly efficient DNA repair machineries, the nucleotide excision repair (NER) system amongst these. Although NER is of central importance to DNA repair in M. tuberculosis, our understanding of the processes in this species is limited. The conserved UvrABC endonuclease represents the multi-enzymatic core in bacterial NER, where the UvrA ATPase provides the DNA lesion-sensing function. The herein reported genetic analysis demonstrates that M. tuberculosis UvrA is important for the repair of nitrosative and oxidative DNA damage. Moreover, our biochemical and structural characterization of recombinant M. tuberculosis UvrA contributes new insights into its mechanism of action. In particular, the structural investigation reveals an unprecedented conformation of the UvrB binding domain that we propose to be of functional relevance. Taken together, our data suggest UvrA as a potential target for the development of novel anti tubercular agents and provide a biochemical framework for the identification of small-molecule inhibitors interfering with the NER activity in M. tuberculosis. PMID- 21622957 TI - DARIO: a ncRNA detection and analysis tool for next-generation sequencing experiments. AB - Small non-coding RNAs (ncRNAs) such as microRNAs, snoRNAs and tRNAs are a diverse collection of molecules with several important biological functions. Current methods for high-throughput sequencing for the first time offer the opportunity to investigate the entire ncRNAome in an essentially unbiased way. However, there is a substantial need for methods that allow a convenient analysis of these overwhelmingly large data sets. Here, we present DARIO, a free web service that allows to study short read data from small RNA-seq experiments. It provides a wide range of analysis features, including quality control, read normalization, ncRNA quantification and prediction of putative ncRNA candidates. The DARIO web site can be accessed at http://dario.bioinf.uni-leipzig.de/. PMID- 21622958 TI - psRNATarget: a plant small RNA target analysis server. AB - Plant endogenous non-coding short small RNAs (20-24 nt), including microRNAs (miRNAs) and a subset of small interfering RNAs (ta-siRNAs), play important role in gene expression regulatory networks (GRNs). For example, many transcription factors and development-related genes have been reported as targets of these regulatory small RNAs. Although a number of miRNA target prediction algorithms and programs have been developed, most of them were designed for animal miRNAs which are significantly different from plant miRNAs in the target recognition process. These differences demand the development of separate plant miRNA (and ta siRNA) target analysis tool(s). We present psRNATarget, a plant small RNA target analysis server, which features two important analysis functions: (i) reverse complementary matching between small RNA and target transcript using a proven scoring schema, and (ii) target-site accessibility evaluation by calculating unpaired energy (UPE) required to 'open' secondary structure around small RNA's target site on mRNA. The psRNATarget incorporates recent discoveries in plant miRNA target recognition, e.g. it distinguishes translational and post transcriptional inhibition, and it reports the number of small RNA/target site pairs that may affect small RNA binding activity to target transcript. The psRNATarget server is designed for high-throughput analysis of next-generation data with an efficient distributed computing back-end pipeline that runs on a Linux cluster. The server front-end integrates three simplified user-friendly interfaces to accept user-submitted or preloaded small RNAs and transcript sequences; and outputs a comprehensive list of small RNA/target pairs along with the online tools for batch downloading, key word searching and results sorting. The psRNATarget server is freely available at http://plantgrn.noble.org/psRNATarget/. PMID- 21622959 TI - A novel bioinformatics pipeline for identification and characterization of fusion transcripts in breast cancer and normal cell lines. AB - SnowShoes-FTD, developed for fusion transcript detection in paired-end mRNA-Seq data, employs multiple steps of false positive filtering to nominate fusion transcripts with near 100% confidence. Unique features include: (i) identification of multiple fusion isoforms from two gene partners; (ii) prediction of genomic rearrangements; (iii) identification of exon fusion boundaries; (iv) generation of a 5'-3' fusion spanning sequence for PCR validation; and (v) prediction of the protein sequences, including frame shift and amino acid insertions. We applied SnowShoes-FTD to identify 50 fusion candidates in 22 breast cancer and 9 non-transformed cell lines. Five additional fusion candidates with two isoforms were confirmed. In all, 30 of 55 fusion candidates had in-frame protein products. No fusion transcripts were detected in non-transformed cells. Consideration of the possible functions of a subset of predicted fusion proteins suggests several potentially important functions in transformation, including a possible new mechanism for overexpression of ERBB2 in a HER-positive cell line. The source code of SnowShoes-FTD is provided in two formats: one configured to run on the Sun Grid Engine for parallelization, and the other formatted to run on a single LINUX node. Executables in PERL are available for download from our web site: http://mayoresearch.mayo.edu/mayo/research/biostat/stand-alone-packages.cfm. PMID- 21622960 TI - Replication enhancer elements within the open reading frame of tick-borne encephalitis virus and their evolution within the Flavivirus genus. AB - We provide experimental evidence of a replication enhancer element (REE) within the capsid gene of tick-borne encephalitis virus (TBEV, genus Flavivirus). Thermodynamic and phylogenetic analyses predicted that the REE folds as a long stable stem-loop (designated SL6), conserved among all tick-borne flaviviruses (TBFV). Homologous sequences and potential base pairing were found in the corresponding regions of mosquito-borne flaviviruses, but not in more genetically distant flaviviruses. To investigate the role of SL6, nucleotide substitutions were introduced which changed a conserved hexanucleotide motif, the conformation of the terminal loop and the base-paired dsRNA stacking. Substitutions were made within a TBEV reverse genetic system and recovered mutants were compared for plaque morphology, single-step replication kinetics and cytopathic effect. The greatest phenotypic changes were observed in mutants with a destabilized stem. Point mutations in the conserved hexanucleotide motif of the terminal loop caused moderate virus attenuation. However, all mutants eventually reached the titre of wild-type virus late post-infection. Thus, although not essential for growth in tissue culture, the SL6 REE acts to up-regulate virus replication. We hypothesize that this modulatory role may be important for TBEV survival in nature, where the virus circulates by non-viraemic transmission between infected and non-infected ticks, during co-feeding on local rodents. PMID- 21622961 TI - CoPub update: CoPub 5.0 a text mining system to answer biological questions. AB - In this article, we present CoPub 5.0, a publicly available text mining system, which uses Medline abstracts to calculate robust statistics for keyword co occurrences. CoPub was initially developed for the analysis of microarray data, but we broadened the scope by implementing new technology and new thesauri. In CoPub 5.0, we integrated existing CoPub technology with new features, and provided a new advanced interface, which can be used to answer a variety of biological questions. CoPub 5.0 allows searching for keywords of interest and its relations to curated thesauri and provides highlighting and sorting mechanisms, using its statistics, to retrieve the most important abstracts in which the terms co-occur. It also provides a way to search for indirect relations between genes, drugs, pathways and diseases, following an ABC principle, in which A and C have no direct connection but are connected via shared B intermediates. With CoPub 5.0, it is possible to create, annotate and analyze networks using the layout and highlight options of Cytoscape web, allowing for literature based systems biology. Finally, operations of the CoPub 5.0 Web service enable to implement the CoPub technology in bioinformatics workflows. CoPub 5.0 can be accessed through the CoPub portal http://www.copub.org. PMID- 21622962 TI - Rosetta FlexPepDock web server--high resolution modeling of peptide-protein interactions. AB - Peptide-protein interactions are among the most prevalent and important interactions in the cell, but a large fraction of those interactions lack detailed structural characterization. The Rosetta FlexPepDock web server (http://flexpepdock.furmanlab.cs.huji.ac.il/) provides an interface to a high resolution peptide docking (refinement) protocol for the modeling of peptide protein complexes, implemented within the Rosetta framework. Given a protein receptor structure and an approximate, possibly inaccurate model of the peptide within the receptor binding site, the FlexPepDock server refines the peptide to high resolution, allowing full flexibility to the peptide backbone and to all side chains. This protocol was extensively tested and benchmarked on a wide array of non-redundant peptide-protein complexes, and was proven effective when applied to peptide starting conformations within 5.5 A backbone root mean square deviation from the native conformation. FlexPepDock has been applied to several systems that are mediated and regulated by peptide-protein interactions. This easy to use and general web server interface allows non-expert users to accurately model their specific peptide-protein interaction of interest. PMID- 21622963 TI - Predicting cardiac output responses to passive leg raising by a PEEP-induced increase in central venous pressure, in cardiac surgery patients. AB - BACKGROUND: Changes in central venous pressure (CVP) rather than absolute values may be used to guide fluid therapy in critically ill patients undergoing mechanical ventilation. We conducted a study comparing the changes in the CVP produced by an increase in PEEP and stroke volume variation (SVV) as indicators of fluid responsiveness. Fluid responsiveness was assessed by the changes in cardiac output (CO) produced by passive leg raising (PLR). METHODS: In 20 fully mechanically ventilated patients after cardiac surgery, PEEP was increased +10 cm H2O for 5 min followed by PLR. CVP, SVV, and thermodilution CO were measured before, during, and directly after the PEEP challenge and 30 degrees PLR. The CO increase >7% upon PLR was used to define responders. RESULTS: Twenty patients were included; of whom, 10 responded to PLR. The increase in CO by PLR directly related (r=0.77, P<0.001) to the increase in CVP by PEEP. PLR responsiveness was predicted by the PEEP-induced increase in CVP [area under receiver-operating characteristic (AUROC) curve 0.99, P<0.001] and by baseline SVV (AUROC 0.90, P=0.003). The AUROC's for dCVP and SVV did not differ significantly (P=0.299). CONCLUSIONS: Our data in mechanically ventilated, cardiac surgery patients suggest that the newly defined parameter, PEEP-induced CVP changes, like SVV, appears to be a good parameter to predict fluid responsiveness. PMID- 21622964 TI - Habitual exercise increases resistance of lymphocytes to oxidant-induced DNA damage by upregulating expression of antioxidant and DNA repairing enzymes. AB - The underlying mechanisms of adaptation from staying physically active are not completely revealed. This study examined the effects of 8 and 20 weeks of habitual voluntary exercise on the susceptibility of lymphocytes to oxidant induced DNA damage, antioxidant enzyme activities in cardiac and skeletal muscles, and circulatory antioxidant profile. Forty young adult rats were randomly assigned to sedentary control and exercise groups for an experimental period of 8 or 20 weeks. Animals assigned to exercise groups were subjected to 24 h daily free access to an in-cage running wheel with circumference of 1.19 m. A magnetic digital counter was attached to the running wheel to record daily exercise distance run by the animals. Control rats were housed in cages without a running wheel, located next to the exercised animals. Body weight and food intake were recorded weekly. After the experimental periods of 8 and 20 weeks, blood, left ventricle, soleus and plantaris muscles were collected for analysis. No significant difference was found in plasma total antioxidant capacity between exercised and control animals in the 8 and 20 week groups according to our ferric reducing/antioxidant power (FRAP) analysis. However, modified FRAP for ascorbic acid (FRASC) analysis indicated that plasma ascorbic acid content was significantly increased by 46 and 34% in 8 and 20 week exercise groups, respectively, when compared with the corresponding control groups. Superoxide dismutase (SOD) activity was significantly elevated by 39% in erythrocytes of animals exercised for 8 weeks relative to control animals. In the 20 week exercise group, Glutathione peroxidase (GPx) activity in ventricle and plantaris was significantly upregulated by 477 and 290%, respectively, relative to control values. As demonstrated by comet assay, the oxidant-induced DNA damage was significantly reduced by 21 and 45% in lymphocytes of animals exercised for 8 and 20 weeks, respectively, when compared with the corresponding control lymphocytes. Our qRT-PCR analysis showed that the transcript expression of SOD2 was significantly elevated by 939% in lymphocytes of animals exercised for 8 weeks relative to control animals. Increased expressions of SOD2 (by 19%), catalase (25%), APEX nuclease (multifunctional DNA repair enzyme) 1 (APEX1; 46%), Protein kinase, DNA-activated, catalytic polypeptide (Prkdc; 9%) and O-6-methylguanine DNA methyltransferase (Mgmt; 26%) were found in lymphocytes of animals exercised for 20 weeks relative to control rats. These results demonstrate that habitual exercise confers increased resistance of lymphocytes to oxidant-induced DNA damage, and this protective effect is possibly attributed to the regular exercise induced elevated expression of antioxidant and DNA repairing enzymes. PMID- 21622965 TI - Left ventricle structural remodelling in the prediabetic Goto-Kakizaki rat. AB - This study tested the hypothesis that experimental prediabetes can elicit structural remodelling in the left ventricle (LV). Left ventricles isolated from 8-week-old male Goto-Kakizaki (GK) rats and age-matched male Wistar control rats were used to assess remodelling changes and underlying transforming growth factor beta1 (TGFbeta1) activity, prohypertrophic Akt-p70S6K1 signalling and gene expression profile of the extracellular matrix (ECM) using histological, immunohistochemical, immunoblotting and quantitative gene expression analyses. Prediabetes in GK rats was confirmed by impaired glucose tolerance and modestly elevated fasting blood glucose. Left ventricle remodelling in the GK rat presented with marked hypertrophy of cardiomyocytes and increased ECM deposition that together translated into increased heart size in the absence of ultrastructural changes or fibre disarray. Molecular derangements underlying this phenotype included recapitulation of the fetal gene phenotype markers B-type natriuretic peptide and alpha-skeletal muscle actin, activation of the Akt p70S6K1 pathway and altered gene expression profile of key components (collagen 1alpha and fibronectin) and modulators of the ECM (matrix metalloproteinases 2 and 9 and connective tissue growth factor). These changes were correlated with parallel findings of increased TGFbeta1 transcription and activation in the LV and elevated active TGFbeta1 in plasma of GK rats compared with control animals (Student's t test, P < 0.05 versus age-matched Wistar control animals for all parameters). This is the first report to describe LV structural remodelling in experimental prediabetes. The results suggest that ventricular decompensation pathognomonic of advanced diabetic cardiomyopathy may have possible origins in profibrotic and prohypertrophic mechanisms triggered before the onset of type 2 diabetes mellitus. PMID- 21622966 TI - Arachidonic acid products in airway nociceptor activation during acute lung injury. AB - We have reported that airway nociceptors [C fibre receptors (CFRs) and high threshold Adelta fibre receptors (HTARs)] are activated during oleic acid (OA) induced acute lung injury. In the present studies, we tested the hypothesis that this nociceptor activation is mediated by arachidonic acid products. In anaesthetized, open-chest, mechanically ventilated rabbits, we examined the response of the nociceptors to intravenous injection of OA before and after blocking the cyclo-oxygenase pathways with indomethacin. Pretreatment with indomethacin (20 mg kg(-1)) decreased the background activities of both CFRs (from 0.48 +/- 0.12 to 0.25 +/- 0.08 impulses/s, n = 7, P < 0.05) and HTARs (from 0.54 +/- 0.14 to 0.23 +/- 0.08 impulses/s, n = 10, P < 0.01). It also blocked the response of the nociceptors to OA. Likewise, pretreatment with thromboxane synthase inhibitor (ketoconazole) also blocked the nociceptor response to OA. In addition, local microinjection or intravenous injection of a thromboxane mimetic stimulated CFRs and HTARs. The present results clearly indicate that arachidonic acid metabolites mediate airway nociceptor activation during OA-induced acute lung injury and suggest that thromboxane may be a key mediator. PMID- 21622967 TI - Muscle glycogen resynthesis, signalling and metabolic responses following acute exercise in exercise-trained pigs carrying the PRKAG3 mutation. AB - Hampshire pigs carrying the PRKAG3 mutation in the AMP-activated protein kinase (AMPK) gamma3 subunit exhibit excessive skeletal muscle glycogen storage and an altered glycogen synthesis signalling response following exercise. AMPK plays an important role as a regulator of carbohydrate and fat metabolism in mammalian cells. Exercise-trained muscles are repeatedly exposed to glycogen degradation and resynthesis, to which the signalling pathways adapt. The aim of this study was to examine the effect of acute exercise on glycogen synthesis signalling pathways, and the levels of insulin and other substrates in blood in exercise trained pigs with and without the PRKAG3 mutation. After 5 weeks of training, pigs performed two standardized treadmill exercise tests, and skeletal muscle biopsies were obtained immediately after exercise and 3 h postexercise in the first test, and 6 h postexercise in the second test. The PRKAG3 mutation carriers had higher glycogen storage, and resynthesis of glycogen was faster after 3 h but not after 6 h of recovery. Alterations in the concentrations of insulin, glucose, lactate and free fatty acids after exercise did not differ between the genotypes. The carriers showed a lower expression of AMPK and increased phosphorylation of Akt Ser(473) after exercise, compared with non-carriers. Acute exercise stimulated the phosphorylation of AS160 in both genotypes, and the phosphorylation of GSK3alpha Ser(21) and ACC Ser(79) in the non-carriers. In conclusion, exercise-trained pigs carrying the PRKAG3 mutation show an altered Akt and AMPK signalling response to acute exercise, indicating that glucose metabolism is associated with faster resynthesis of muscle glycogen in this group. PMID- 21622968 TI - Accelerated metacarpal bone mineral density loss is associated with radiographic progressive hand osteoarthritis. AB - OBJECTIVE: To study the association between metacarpal bone mineral density (BMD) loss and progressive hand osteoarthritis (OA) over 2 years. METHODS: Using the Kellgren-Lawrence (KL) grading scale and the Osteoarthritis Research Society International Atlas, standardised hand radiographs of 181 patients with primary OA at multiple sites (mean age 60 years, 80% women, mean body mass index 27 kg/m(2)) were assessed for hand OA at baseline (KL >= 2 in two or more hand joints) and progressive hand OA over 2 years (>= 1 point increase in total osteophyte and joint space narrowing score in patients with hand OA at baseline). Changes in BMD were measured over 2 years in metacarpals 2-4 by digital x-ray radiogrammetry. Accelerated BMD loss was defined as loss of >3 mg/cm(2)/year. Logistic regression analyses were performed to assess the associations between BMD loss and progressive hand OA. RESULTS: The baseline prevalence of hand OA was 68% and, after 2 years, 32% of these patients had progressive hand OA. Accelerated BMD loss was present in 79% of the patients with progressive hand OA compared with 60% and 57% of the patients with non-progressive hand OA and no hand OA, respectively. BMD loss was independently associated with progressive hand OA compared with non-progressive hand OA with a RR (95% CI) of 2.1 (1.1 to 4.3). CONCLUSION: Accelerated metacarpal BMD loss is associated with progressive hand OA over a period of 2 years; knowledge of common mechanisms may lead to development of therapeutic interventions for hand OA. PMID- 21622969 TI - Rates and predictors of radiographic sacroiliitis progression over 2 years in patients with axial spondyloarthritis. AB - OBJECTIVE: To assess the progression of radiographic sacroiliitis in a cohort of patients with early axial spondyloarthritis over a period of 2 years and to explore predictors of progression. METHODS: 210 patients with axial spondyloarthritis from the German Spondyloarthritis Inception Cohort have been selected for this analysis based on availability of radiographs at baseline and after 2 years of follow-up. Radiographs were centrally digitised and the sacroiliac joints were scored independently according to the grading system of the modified New York criteria for ankylosing spondylitis (AS) by two trained readers. The readers scored both time points simultaneously but were blinded for the time point and for all clinical data. RESULTS: 115 patients (54.8%) fulfilled the modified New York criteria for AS in their radiographic part in the opinion of both readers at baseline, while 95 patients (45.2%) were classified as non radiographic axial spondyloarthritis. More patients with non-radiographic spondyloarthritis (10.5%) compared with AS (4.4%) showed an estimated 'true' progression by at least one grade according to both readers, although the difference between the two groups was statistically non-significant. The rate of progression from non-radiographic axial spondyloarthritis to AS was 11.6% over 2 years. An elevated level of C-reactive protein (CRP) at baseline was a strong positive predictor of radiographic sacroiliitis progression in non-radiographic axial spondyloarthritis and AS (OR 3.65 and 5.08, respectively, p<0.05). CONCLUSION: Progression of radiographic sacroiliitis by at least one grade after 2 years occurs only in a small percentage of patients with early axial spondyloarthritis. An elevated level of CRP was found to be a strong positive predictor of sacroiliitis progression. PMID- 21622970 TI - Monosodium urate monohydrate crystals inhibit osteoblast viability and function: implications for development of bone erosion in gout. AB - BACKGROUND: Bone erosion is a common manifestation of chronic tophaceous gout. OBJECTIVES: To investigate the effects of monosodium urate monohydrate (MSU) crystals on osteoblast viability and function. METHODS: The MTT assay and flow cytometry were used to assess osteoblast cell viability in the MC3T3-E1 and ST2 osteoblast-like cell lines, and primary rat and primary human osteoblasts cultured with MSU crystals. Quantitative real-time PCR and von Kossa stained mineralised bone formation assays were used to assess the effects of MSU crystals on osteoblast differentiation using MC3T3-E1 cells. The numbers of osteoblasts and bone lining cells were quantified in bone samples from patients with gout. RESULTS: MSU crystals rapidly reduced viability in all cell types in a dose dependent manner. The inhibitory effect on cell viability was independent of crystal phagocytosis and was not influenced by differing crystal length or addition of serum. Long-term culture of MC3T3-E1 cells with MSU crystals showed a reduction in mineralisation and decreased mRNA expression of genes related to osteoblast differentiation such as Runx2, Sp7 (osterix), Ibsp (bone sialoprotein), and Bglap (osteocalcin). Fewer osteoblast and lining cells were present on bone directly adjacent to gouty tophus than bone unaffected by tophus in patients with gout. CONCLUSIONS: MSU crystals have profound inhibitory effects on osteoblast viability and differentiation. These data suggest that bone erosion in gout occurs at the tophus-bone interface through alteration of physiological bone turnover, with both excessive osteoclast formation, and reduced osteoblast differentiation from mesenchymal stem cells. PMID- 21622971 TI - Treatment for hepatitis C virus with pegylated interferon-alpha plus ribavirin induces anti-atherogenic effects on cardiovascular risk biomarkers in HIV infected and -uninfected patients. AB - OBJECTIVES: We investigated the influence of hepatitis C virus (HCV) therapy with pegylated interferon-alpha plus ribavirin on cardiovascular disease risk through the serial measurement of several laboratory markers in HCV-monoinfected and HCV/HIV-coinfected patients. METHODS: In a longitudinal study, biomarkers of inflammation, coagulation and oxidative stress were measured during and after therapy. RESULTS: A total of 56 patients were included; 32 (57.1%) were HCV/HIV coinfected and 24 (42.9%) were HCV monoinfected. Compared with baseline, during HCV therapy there was a significant decrease in the concentrations of matrix metalloproteinase-9 (P < 0.001), intercellular cell adhesion molecule-1 (ICAM-1) (P <= 0.01) and oxidized low-density lipoproteins (P = 0.002). In contrast, levels of vascular cell adhesion molecule-1 (VCAM-1), monocyte chemotactic protein-1 and fibrinogen increased during treatment. After treatment discontinuation, levels of ICAM-1, VCAM-1 and tumour necrosis factor-alpha were significantly lower compared with baseline, a change restricted to patients with sustained virological response. Decreases in transaminases and HCV-RNA from baseline correlated positively with the decrease in ICAM-1 concentration 6 months after treatment discontinuation. Changes in biomarkers were similar in HIV infected and -uninfected patients. CONCLUSIONS: Treatment for HCV induces different changes in several cardiovascular risk biomarkers, most being anti atherogenic effects, although only the anti-atherogenic effects remain after treatment discontinuation in patients with sustained virological response. PMID- 21622972 TI - Wax moth larva (Galleria mellonella): an in vivo model for assessing the efficacy of antistaphylococcal agents. AB - OBJECTIVES: To investigate whether the wax moth larva, Galleria mellonella, is a suitable host for assessing the in vivo efficacy of antistaphylococcal agents against Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) infections. METHODS: Wax moth larvae were infected with increasing doses of S. aureus to investigate the effect of inoculum size on larval survival. In addition, infected wax moth larvae were treated with daptomycin, penicillin or vancomycin to examine whether these agents were effective against S. aureus and MRSA infections in vivo. RESULTS: Increasing inoculum doses of live S. aureus cells resulted in greater larval mortality, but heat-killed bacteria and cell free culture filtrates had no detrimental effects on survival. Larval mortality rate also depended on the post-inoculation incubation temperature. After larvae were infected with S. aureus, larval survival was enhanced by administering the antistaphylococcal antibiotics daptomycin or vancomycin. Larval survival increased with increasing doses of the antibiotics. Moreover, penicillin improved survival of larvae infected with a penicillin-susceptible methicillin-susceptible S. aureus (MSSA) strain, but it was ineffective at similar doses in larvae infected with MRSA (penicillin resistant). Daptomycin and vancomycin were also effective when administered to the larvae prior to infection with bacteria. CONCLUSIONS: This is the first report to demonstrate that antibiotics are effective in the wax moth larva model for the treatment of infections caused by Gram-positive bacteria. The new wax moth larva model is a useful preliminary model for assessing the in vivo efficacy of candidate antistaphylococcal agents before proceeding to mammalian studies, which may reduce animal experimentation and expense. PMID- 21622973 TI - Daptomycin resistance mechanisms in clinically derived Staphylococcus aureus strains assessed by a combined transcriptomics and proteomics approach. AB - OBJECTIVES: The development of daptomycin resistance in Staphylococcus aureus is associated with clinical treatment failures. The mechanism(s) of such resistance have not been clearly defined. METHODS: We studied an isogenic daptomycin susceptible (DAP(S)) and daptomycin-resistant (DAP(R)) S. aureus strain pair (616; 701) from a patient with relapsing endocarditis during daptomycin treatment, using comparative transcriptomic and proteomic techniques. RESULTS: Minor differences in the genome content were found between strains by DNA hybridization. Transcriptomic analyses identified a number of genes differentially expressed in important functional categories: cell division; metabolism of bacterial envelopes; and global regulation. Of note, the DAP(R) isolate exhibited reduced expression of the major cell wall autolysis gene coincident with the up-regulation of genes involved in cell wall teichoic acid production. Using quantitative (q)RT-PCR on the gene cadre putatively involved in cationic peptide resistance, we formulated a putative regulatory network compatible with microarray data sets, mainly implicating bacterial envelopes. Of interest, qRT-PCR of this same gene cadre from two distinct isogenic DAP(S)/DAP(R) clinical strain pairs revealed evidence of other strain-dependent networks operative in the DAP(R) phenotype. Comparative proteomics of 616 versus 701 revealed a differential abundance of proteins in various functional categories, including cell wall-associated targets and biofilm formation proteins. Phenotypically, strains 616 and 701 showed major differences in their ability to develop bacterial biofilms in the presence of the antibacterial lipid, oleic acid. CONCLUSIONS: Compatible with previous in vitro observations, in vivo acquired DAP(R) in S. aureus is a complex, multistep phenomenon involving: (i) strain-dependent phenotypes; (ii) transcriptome adaptation; and (iii) modification of the lipid and protein contents of cellular envelopes. PMID- 21622974 TI - Interaction of N-methyl-2-alkenyl-4-quinolones with ATP-dependent MurE ligase of Mycobacterium tuberculosis: antibacterial activity, molecular docking and inhibition kinetics. AB - OBJECTIVES: The aim of this study was to comprehensively evaluate the antibacterial activity and MurE inhibition of a set of N-methyl-2-alkenyl-4 quinolones found to inhibit the growth of fast-growing mycobacteria. METHODS: Using the spot culture growth inhibition assay, MICs were determined for Mycobacterium tuberculosis H(37)Rv, Mycobacterium bovis BCG and Mycobacterium smegmatis mc(2)155. MICs were determined for Mycobacterium fortuitum, Mycobacterium phlei, methicillin-resistant Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa using microplate dilution assays. Inhibition of M. tuberculosis MurE ligase activity was determined both by colorimetric and HPLC methods. Computational modelling and binding prediction of the quinolones in the MurE structure was performed using Glide. Kinetic experiments were conducted for understanding possible competitive relations of the quinolones with the endogenous substrates of MurE ligase. RESULTS: The novel synthetic N-methyl-2 alkenyl-4-quinolones were found to be growth inhibitors of M. tuberculosis and rapid-growing mycobacteria as well as methicillin-resistant S. aureus, while showing no inhibition for E. coli and P. aeruginosa. The quinolones were found to be inhibitory to MurE ligase of M. tuberculosis in the micromolar range (IC(50) ~40-200 MUM) when assayed either spectroscopically or by HPLC. Computational docking of the quinolones on the published M. tuberculosis MurE crystal structure suggested that the uracil recognition site is a probable binding site for the quinolones. CONCLUSIONS: N-methyl-2-alkenyl-4-quinolones are inhibitors of mycobacterial and staphylococcal growth, and show MurE ligase inhibition. Therefore, they are considered as a starting point for the development of increased affinity MurE activity disruptors. PMID- 21622975 TI - Beneficial effects of combinatorial micronutrition on body fat and atherosclerosis in mice. AB - AIMS: More than two billion people worldwide are deficient in key micronutrients. Single micronutrients have been used at high doses to prevent and treat dietary insufficiencies. Yet the impact of combinations of micronutrients in small doses aiming to improve lipid disorders and the corresponding metabolic pathways remains incompletely understood. Thus, we investigated whether a combination of micronutrients would reduce fat accumulation and atherosclerosis in mice. METHODS AND RESULTS: Lipoprotein receptor-null mice fed with an original combination of micronutrients incorporated into the daily chow showed reduced weight gain, body fat, plasma triglycerides, and increased oxygen consumption. These effects were achieved through enhanced lipid utilization and reduced lipid accumulation in metabolic organs and were mediated, in part, by the nuclear receptor PPARalpha. Moreover, the micronutrients partially prevented atherogenesis when administered early in life to apolipoprotein E-null mice. When the micronutrient treatment was started before conception, the anti-atherosclerotic effect was stronger in the progeny. This finding correlated with decreased post-prandial triglyceridaemia and vascular inflammation, two major atherogenic factors. CONCLUSION: Our data indicate beneficial effects of a combination of micronutritients on body weight gain, hypertriglyceridaemia, liver steatosis, and atherosclerosis in mice, and thus our findings suggest a novel cost-effective combinatorial micronutrient based strategy worthy of being tested in humans. PMID- 21622976 TI - The impact of VEGF and bFGF on vascular stereomorphology in the context of angiogenic neo-arborisation after vascular induction. AB - The aim of this in vivo study was to gather quantitative information on the three dimensional morphology of a new vascular network under the influence of angioactive growth factors. For this purpose, the arteriovenous loop model was used in 10 Lewis rats to generate a bioartificial vascular assembly by means of vascular induction. In this model, an isolated organoid is created in the medial thigh of the animal by methods of tissue engineering. A fibrin gel containing vascular endothelial growth factor (VEGF(165)) and basic fibroblastic growth factor (bFGF) was used as a matrix in the effect group (GF+). Fibrin matrices devoid of growth factors were used as controls (GF-). A microvascular replica of the organoid was created by means of corrosion casting and the network was investigated on stereo-paired images obtained by scanning electron microscopy. Vectors of intercapillary and interbranching distances as well as the diameter of the pores in the intussusceptive events diameter and the ratio of sprouting versus intussusceptive angiogenic events were compared in the two groups. The results were highly significant. In the GF+ group there were more profound three dimensional morphological traits of angiogenesis, whereas advanced neovascularisation in the phase of remodelling was demonstrated by a higher incidence of intussusception, compared to control. These results illustrate the importance of morphological studies with focus on the generation of three dimensional vascular networks. PMID- 21622977 TI - Sequence capture and next generation resequencing of the MHC region highlights potential transplantation determinants in HLA identical haematopoietic stem cell transplantation. AB - How cells coordinate the immune system activities is important for potentially life-saving organ or stem cell transplantations. Polymorphic immunoregulatory genes, many of them located in the human major histocompatibility complex, impact the process and assure the proper execution of tolerance-versus-activity mechanisms. In haematopoietic stem cell transplantation, on the basis of fully human leukocyte antigen (HLA)-matched donor-recipient pairs, adverse effects like graft versus leukaemia and graft versus host are observed and difficult to handle. So far, high-resolution HLA typing was performed with Sanger sequencing, but for methodological reasons information on additional immunocompetent major histocompatibility complex loci has not been revealed. Now, we have used microarray sequence capture and targeted enrichment combined with next generation pyrosequencing for 3.5 million base pair human major histocompatibility complex resequencing in a clinical transplant setting and describe 3025 variant single nucleotide polymorphisms, insertions and deletions among recipient and donor in a single sequencing experiment. Taken together, the presented data show that sequence capture and massively parallel pyrosequencing can be used as a new tool for risk assessment in the setting of allogeneic stem cell transplantation. PMID- 21622978 TI - Significance of (sub)clinical thyroid dysfunction and thyroid autoimmunity before conception and in early pregnancy: a systematic review. AB - BACKGROUND: Thyroid dysfunction and thyroid autoimmunity are prevalent among women of reproductive age and are associated with adverse pregnancy outcomes. Preconception or early pregnancy screening for thyroid dysfunction has been proposed but is not widely accepted. We conducted a systematic review of the literature on the clinical significance of thyroid dysfunction and thyroid autoimmunity before conception and in early pregnancy. METHODS: Relevant studies were identified by searching Medline, EMBASE and the Cochrane Controlled Trials Register. RESULTS: From a total of 14 208 primary selected titles, 43 articles were included for the systematic review and 38 were appropriate for meta analyses. No articles about hyperthyroidism were selected. Subclinical hypothyroidism in early pregnancy, compared with normal thyroid function, was associated with the occurrence of pre-eclampsia [odds ratio (OR) 1.7, 95% confidence interval (CI) 1.1-2.6] and an increased risk of perinatal mortality (OR 2.7, 95% CI 1.6-4.7). In the meta-analyses, the presence of thyroid antibodies was associated with an increased risk of unexplained subfertility (OR 1.5, 95% CI 1.1-2.0), miscarriage (OR 3.73, 95% CI 1.8-7.6), recurrent miscarriage (OR 2.3, 95% CI 1.5-3.5), preterm birth (OR 1.9, 95% CI 1.1-3.5) and maternal post-partum thyroiditis (OR 11.5, 95% CI 5.6-24) when compared with the absence of thyroid antibodies. CONCLUSIONS: Pregnant women with subclinical hypothyroidism or thyroid antibodies have an increased risk of complications, especially pre-eclampsia, perinatal mortality and (recurrent) miscarriage. Future research, within the setting of clinical trials, should focus on the potential health gain of identification, and effect of treatment, of thyroid disease on pregnancy outcome. PMID- 21622979 TI - Timing and potential mechanisms of new conduction abnormalities during the implantation of the Medtronic CoreValve System in patients with aortic stenosis. AB - AIMS: New-onset left bundle branch block (LBBB) and complete atrioventricular block (AV3B) frequently occur following transcatheter aortic valve implantation (TAVI). We sought to determine the timing and potential mechanisms of new conduction abnormalities (CAs) during TAVI, using the Medtronic CoreValve System (MCS). METHODS AND RESULTS: Sixty-five consecutive patients underwent TAVI with continuous 12-lead ECG analysis. New CAs were defined by the occurrence of LBBB, RBBB, and/or AV3B after the following pre-defined time points: (i) crossing of valve with stiff wire, (ii) positioning of balloon catheter in the aortic annulus, (iii) balloon valvuloplasty, (iv) positioning of MCS in the left ventricular outflow tract (LVOT), (v) expansion of MCS, (vi) removal of all catheters. A new CA occurred during TAVI in 48 patients (74%) and after TAVI in 5 (8%). Of the 48 patients with procedural CAs, a single new CA occurred in 43 patients (90%) and two types of CAs in 5 (10%). A new LBBB was seen in 40 patients (83%), AV3B in 9 (19%), and RBBB in 4 (8%). The new CA first occurred-in descending order of frequency-after balloon valvuloplasty in 22 patients (46%), MCS expansion in 14 (29%), MCS positioning in 6 (12%), positioning of balloon catheter in 3 (6%), wire-crossing of aortic valve in 2 (4%), and after catheter removal in 1 patient (2%). Patients who developed a new CA during balloon valvuloplasty had a significantly higher balloon/annulus ratio than those who did not (1.10+/-0.10 vs. 1.03+/-0.11, P=0.030). No such relationship was found with the valve/annulus ratio. CONCLUSION: Transcatheter aortic valve implantation with the MCS was associated with new CAs in 82% of which more than half occurred before the actual valve implantation. It remains to be elucidated by dedicated studies whether new CAs can be reduced by appropriate balloon sizing-a precept that also holds for valve size given the observed directional signal of the valve size/aortic annulus ratio. PMID- 21622981 TI - Pharmacotherapy at the end-of-life. AB - Older people reaching end-of-life status are particularly at risk from inter related adverse effects of pharmacotherapy, including polypharmacy, inappropriate medications and adverse drug events. These adverse effects of pharmacotherapy may be highly detrimental, as well as highly expensive. End-of-life pharmacotherapy is sometimes perceived to be complex and challenging, probably unnecessarily. This relates in part to the poorly developed evidence base and lack of high quality research in this area. In this article, we deal with some of the key issues relating to pharmacotherapy in end-of-life patients, namely (i) the guiding principles of drug selection, (ii) the main drugs and drug classes that are best avoided, (iii) the benefits of 'oligopharmacy' (i.e. deliberate avoidance of polypharmacy) in end-of-life patients. PMID- 21622980 TI - Early drop in systolic blood pressure and worsening renal function in acute heart failure: renal results of Pre-RELAX-AHF. AB - AIMS: We aimed to determine the relation between baseline systolic blood pressure (SBP), change in SBP, and worsening renal function (WRF) in acute heart failure (AHF) patients enrolled in the Pre-RELAX-AHF trial. METHODS AND RESULTS: The Pre RELAX-AHF study enrolled 234 patients within 16 h of admission (median 7 h) for AHF and randomized them to relaxin given intravenous (i.v.) for 48 h or placebo. Blood pressure was measured at baseline, at 3, 6, 9, 12, 24, 36, and 48 h and at 3, 4, and 5 days after enrolment. Worsening renal function was defined as a serum creatinine increase of >=0.3 mg/dL by Day 5. Worsening renal function was found in 68 of the 225 evaluable patients (30%). Patients with WRF were older (73.5 +/- 9.4 vs. 69.1 +/- 10.6 years; P= 0.003), had a higher baseline SBP (147.3 +/- 19.9 vs. 140.8 +/- 16.7 mmHg; P= 0.01), and had a greater early drop in SBP (37.9 +/- 16.0 vs. 31.4 +/- 12.2 mmHg; P= 0.004). In a multivariable model, higher age, higher baseline creatinine, and a greater early drop in SBP, but not baseline SBP, remained independent predictors of WRF. Furthermore, WRF was associated with a higher Day 60 (P= 0.01), and Day 180 (P= 0.003) mortality. CONCLUSIONS: Worsening renal function in hospitalized AHF patients is related to a poor clinical outcome and is predicted by a greater early drop in SBP. Trial registration clinicaltrials.gov identifier NCT00520806. PMID- 21622982 TI - Genetic dissection of late-life fertility in Caenorhabditis elegans. AB - The large post-reproductive life span reported for the free-living hermaphroditic nematode, Caenorhabditis elegans, which lives for about 10 days after its 5-day period of self-reproduction, seems at odds with evolutionary theory. Species with long post-reproductive life spans such as mammals are sometimes explained by a need for parental care or transfer of information. This does not seem a suitable explanation for C elegans. Previous reports have shown that C elegans can regain fertility when mated after the self-fertile period but did not report the functional limits. Here, we report the functional life span of the C elegans germ line when mating with males. We show that C elegans can regain fertility late in life (significantly later than in previous reports) and that the end of this period corresponds quite well to its 3-week total life span. Genetic analysis reveals that late-life fertility is controlled by conserved pathways involved with aging and dietary restriction. PMID- 21622983 TI - Age-associated vascular oxidative stress, Nrf2 dysfunction, and NF-{kappa}B activation in the nonhuman primate Macaca mulatta. AB - Aging promotes oxidative stress in vascular endothelial and smooth muscle cells, which contribute to the development of cardiovascular diseases. NF-E2-related factor 2 (Nrf2) is a transcription factor, which is activated by reactive oxygen species in the vasculature of young animals, leading to adaptive upregulation of numerous reactive oxygen species detoxifying and antioxidant genes. The present study was designed to elucidate age-associated changes in the homeostatic role of Nrf2-driven free radical detoxification mechanisms in the vasculature of nonhuman primates. We found that carotid arteries of aged rhesus macaques (Macaca mulatta, age: >=20 years) exhibit significant oxidative stress (as indicated by the increased 8-iso-PGF2alpha and 4-HNE content and decreased glutathione and ascorbate levels) as compared with vessels of young macaques (age:~10 years) that is associated with activation of the redox-sensitive proinflammatory transcription factor, nuclear factor-kappaB. However, age-related oxidative stress does not activate Nrf2 and does not induce Nrf2 target genes (NQO1, GCLC, and HMOX1). In cultured vascular smooth muscle cells (VSMCs) derived from young M mulatta, treatment with H(2)O(2) and high glucose significantly increases transcriptional activity of Nrf2 and upregulates the expression of Nrf2 target genes. In contrast, in cultured vascular smooth muscle cells cells derived from aged macaques, H(2)O(2)- and high glucose-induced Nrf2 activity and Nrf2-driven gene expression are blunted. High glucose-induced H(2)O(2) production was significantly increased in aged vascular smooth muscle cells compared with that in vascular smooth muscle cells from young M mulatta. Taken together, aging is associated with Nrf2 dysfunction in M mulatta arteries, which likely exacerbates age-related cellular oxidative stress, promoting nuclear factor-kappaB activation and vascular inflammation in aging. PMID- 21622984 TI - Phospholipase D mediates nutrient input to mammalian target of rapamycin complex 1 (mTORC1). AB - The mammalian target of rapamycin (mTOR) is a critical sensor of nutritional sufficiency. Although much is known about the regulation of mTOR in response to growth factors, much less is known about the regulation of mTOR in response to nutrients. Amino acids have no impact on the signals that regulate Rheb, a GTPase required for the activation of mTOR complex 1 (mTORC1). Phospholipase D (PLD) generates a metabolite, phosphatidic acid, that facilitates association between mTOR and the mTORC1 co-factor Raptor. We report here that elevated PLD activity in human cancer cells is dependent on both amino acids and glucose and that amino acid- and glucose-induced increases in mTORC1 activity are dependent on PLD. Amino acid- and glucose-induced PLD and mTORC1 activity were also dependent on the GTPases RalA and ARF6 and the type III phosphatidylinositol-3-kinase hVps34. Thus, a key stimulatory event for mTORC1 activation in response to nutrients is the generation of phosphatidic acid by PLD. PMID- 21622985 TI - Renin-angiotensin system blockade and kidney transplantation: a longitudinal cohort study. AB - BACKGROUND: The beneficial effect of angiotensin-converting enzyme inhibitors (ACEI) or angiotensin II receptor blockers (ARB) in kidney transplant recipients on modern immunosuppression is not yet well established. Our objective was to investigate the impact of the use of ACEI/ARB on patient and graft survival in a cohort of kidney transplant recipients. METHODS: A total of 990 patients, who received a single deceased donor kidney at our institution between 1996 and 2005, were included in this longitudinal cohort study. All-cause mortality and death censored graft loss were the primary outcomes. We used traditional time-dependent Cox model (unweighted) and inverse-probability-of-treatment weighting of marginal structural models (weighted Cox model), controlling for time-dependent confounding by indication. RESULTS: A total of 414 patients (42%) received ACEI/ARB through the study period (median duration 14 months, interquartile range 6-40 months). ACEI/ARB use was associated with reduction of risk for mortality in the crude [hazard ratio (HR) 0.627, 95% confidence interval (CI) 0.412-0.953] and adjusted Cox analysis (HR 0.626, 95% CI 0.407-0.963). Similar results were observed after adjusting for confounding by indication (HR 0.629, 95% CI 0.407 0.973). By contrast, ACEI/ARB use was not associated with significant improvement of graft survival after kidney transplantation. CONCLUSION: ACEI/ARB prescription may be suggested as beneficial among multiple medications for reducing mortality in kidney transplant recipients, but its use was not associated with longer graft survival. PMID- 21622986 TI - MR measures of renal perfusion, oxygen bioavailability and total renal blood flow in a porcine model: noninvasive regional assessment of renal function. AB - BACKGROUND: Magnetic resonance imaging (MRI) may be a useful adjunct to current methods of evaluating renal function. MRI is a noninvasive imaging modality that has the ability to evaluate the kidneys regionally, which is lacking in current clinical methods. Other investigators have evaluated renal function with MRI based measurements, such as with techniques to measure cortical and medullary perfusion, oxygen bioavailability and total renal blood flow (TRBF). However, use of all three techniques simultaneously, and therefore the relationships between these MRI-derived functional parameters, have not been reported previously. METHODS: To evaluate the ability of these MRI techniques to track changes in renal function, we scanned 11 swine during a state of hyperperfusion with acetylcholine and a saline bolus and subsequently scanned during a state of hypoperfusion with the prolonged use of isoflurane anesthesia. For each time point, measurements of perfusion, oxygen bioavailability and TRBF were acquired. Measurements of perfusion and oxygen bioavailability were compared with measurements of TRBF for all swine across all time points. RESULTS: Cortical perfusion, cortical oxygen bioavailability, medullary oxygen bioavailability and TRBF significantly increased with the acetylcholine challenge. Cortical perfusion, medullary perfusion, cortical oxygen bioavailability and TRBF significantly decreased during isoflurane anesthesia. Cortical perfusion (Spearman's correlation coefficient = 0.68; P < 1 * 10(-6)) and oxygen bioavailability (Spearman's correlation coefficient = -0.60; P < 0.0001) correlated significantly with TRBF, whereas medullary perfusion and oxygen bioavailability did not correlate with TRBF. CONCLUSIONS: Our results demonstrate expected changes given the pharmacologically induced changes in renal function. Maintenance of the medullary oxygen bioavailability in low blood flow states may reflect the autoregulation particular to this region of the kidney. The ability to non-invasively measure all three parameters of kidney function in a single MRI examination and to evaluate the relationships between these functional parameters is potentially useful for evaluating the state of the human kidneys in situ in future studies. PMID- 21622987 TI - Impact of sirolimus, tacrolimus and mycophenolate mofetil on osteoclastogenesis- implications for post-transplantation bone disease. AB - BACKGROUND: Post-transplantation bone disease is associated with a high degree of morbidity including pain and fractures. Glucocorticoid-induced osteoporosis on top of pre-existing renal osteodystrophy is considered the major pathogenic factor, while the role of non-glucocorticoid immunosuppressants is less well defined. METHODS: In this study, we investigated the influence of sirolimus (SRL) versus calcineurin inhibitor (CI)-based immunosuppressive regimens on biomarkers of bone resorption in renal transplant patients. In addition, the impact of SRL, tacrolimus and mycophenolate mofetil (MMF) on osteoclast activation and function was assessed in cell culture systems. RESULTS: Using this approach, we demonstrated reduced serum levels of bone resorption markers in patients treated with SRL after kidney transplantation compared to a CI-based regimen. In line with this observation, we detected profoundly reduced osteoclast differentiation and subsequently diminished hydroxyapatite resorption in the presence of SRL compared to MMF and tacrolimus in vitro. Moreover, SRL significantly reduced osteoclast precursor proliferation in vitro compared to tacrolimus and led to augmented apoptosis in osteoclast precursors. CONCLUSIONS: Taken together, SRL was shown to inhibit osteoclast formation in vivo and in vitro. SRL thus may have the potential to balance osteoclast promoting effects of glucocorticoids and CI, thereby counteracting the development of accelerated osteoporosis in renal transplant recipients. PMID- 21622989 TI - Extended daily on-line high-volume haemodiafiltration in septic multiple organ failure: a well-tolerated and feasible procedure. AB - BACKGROUND: The outcome of patients with septic multiple organ failure (MOF) remains poor. There are experimental and clinical data indicating a beneficial effect of high-volume haemofiltration. Delivering high-volume therapy is only cost effective using on-line devices because of high costs for additional solution bags in conventional continuous renal replacement therapy (CRRT). We investigated feasibility and effectiveness of extended daily on-line high-volume haemodiafiltration (HDF) with technically maximum convective volume in patients with septic MOF in a pilot study. METHODS: We included 21 consecutive critically ill patients with septic MOF having a mortality risk >50% (SAPS II >50, APACHE II >25). Renal replacement therapy (RRT) was applied with extended daily HDF for 6 23 h using the AK 200 Ultra S dialysis machine in the ultracontrol pre-dilution mode. Dialysate and substitution fluid were prepared on-line. Patients underwent 289 treatments. RESULTS: The mean convective volume was 17.8 +/- 3.7 L/h and 208 +/- 66 mL/kg/h, respectively, median treatment time was 10:15 h/day. Seventeen of 21 patients survived 28 days (81%). The 90-day survival rate was 52% (11/21) versus 19% compared to the survival rate predicted by APACHE II (33.6 mean) and SAPS II (68.6 mean) scores. Haemodynamics improved significantly during the treatment procedures. Material costs per treatment amounted to 35 ?. CONCLUSIONS: Extended daily on-line HDF using maximum convective volume seems to improve the outcome of septic MOF, especially in the early phase. The investigated mode of treatment proved to be feasible, well tolerated and highly cost effective compared to conventional CRRT. At present, this procedure would be applicable at every ICU facility with nephrological support. PMID- 21622988 TI - Meta-analysis of genome-wide linkage scans for renal function traits. AB - BACKGROUND: Several genome scans have explored the linkage of chronic kidney disease phenotypes to chromosomic regions with disparate results. Genome scan meta-analysis (GSMA) is a quantitative method to synthesize linkage results from independent studies and assess their concordance. METHODS: We searched PubMed to identify genome linkage analyses of renal function traits in humans, such as estimated glomerular filtration rate (GFR), albuminuria, serum creatinine concentration and creatinine clearance. We contacted authors for numerical data and extracted information from individual studies. We applied the GSMA nonparametric approach to combine results across 14 linkage studies for GFR, 11 linkage studies for albumin creatinine ratio, 11 linkage studies for serum creatinine and 4 linkage studies for creatinine clearance. RESULTS: No chromosomal region reached genome-wide statistical significance in the main analysis which included all scans under each phenotype; however, regions on Chromosomes 7, 10 and 16 reached suggestive significance for linkage to two or more phenotypes. Subgroup analyses by disease status or ethnicity did not yield additional information. CONCLUSIONS: While heterogeneity across populations, methodologies and study designs likely explain this lack of agreement, it is possible that linkage scan methodologies lack the resolution for investigating complex traits. Combining family-based linkage studies with genome-wide association studies may be a powerful approach to detect private mutations contributing to complex renal phenotypes. PMID- 21622990 TI - Increase of infectious complications in ABO-incompatible kidney transplant recipients--a single centre experience. AB - BACKGROUND: Due to the shortage of deceased donors ABO-incompatible (ABOi) living kidney transplantation has become a popular alternative to deceased kidney transplantation. In recent years, recipient desensitization with a combination of anti-CD20 treatment (rituximab), antigen-specific immunoadsorptions (IA) and intravenous immunoglobulin (IVIG), led to promising short-term and intermediate term results. However, little is known about the impact of this intensified desensitization protocol on the risk of surgical and infectious complications. METHODS: We retrospectively analysed 21 consecutive recipients who underwent ABOi renal transplantation. Pre-transplant desensitization included administration of rituximab (375 mg/m(2)), mycophenolate mofetil (MMF), tacrolimus and prednisolone 4 weeks prior of scheduled transplantation as well as IA and IVIG. Forty-seven patients who underwent ABO-compatible (ABOc) renal transplantation served as the control group. Medical records and electronic databases were reviewed for patient and graft survival, renal function, rate of rejections, viral and bacterial infections as well as for surgical complications (SCs) post-transplantation. RESULTS: All patients showed an immediate graft function. During a mean follow-up of 15.7 +/- 8.3 months (interquartile range 11.9) patient survival was 95 and 98% in the ABOi and ABOc group, respectively. Allograft survival and function, as assessed by serum creatinine levels and calculated glomerular filtration rate at 1 year, did not differ between ABOi and ABOc recipients. Furthermore, the rate of biopsy-proven acute rejections was comparable between the two groups. However, there was a trend towards more SCs within the ABOi group (29 versus 11%, non significant). In addition, the rate of viral infections including cytomegalovirus, Herpes simplex virus, Varicella zoster virus and polyoma virus was significantly increased among the ABOi recipients (50 versus 21%; P = 0.038) despite comparable tacrolimus trough levels and MMF and steroid doses. CONCLUSIONS: Our results, in line with the extended experience of other groups, demonstrate favourable short-term allograft survival and function after ABOi renal transplantation after desensitization with antigen-specific IA, IVIG and rituximab. However, the intensified desensitization was associated with an increased risk of infectious complications. This observation prompted us to briefly escalate the desensitization protocol in ABOi kidney recipients in our centre. PMID- 21622991 TI - Mycophenolate mofetil in low-risk renal transplantation in patients receiving no cyclosporine: a single-centre experience. AB - BACKGROUND: We assess our long-term experience with regards the safety and efficacy of Mycophenolate Mofetil (MMF) in our low risk renal transplant population and compared it retrospectively to Azathioprine (AZA) immunosuppressive regimen. Patients and methods. Between January 1999 and December 2005, 240 renal transplants received MMF as part of their immunosuppressive protocol (MMF group). AZA group of 135 renal transplants was included for comparative analysis (AZA group). Patients received Cyclosporine was excluded from this study. RESULTS: The incidence of biopsy proven 3-month acute rejections was 30 (12.5%) in MMF group and 22 (16%) in AZA group respectively (P = 0.307). Patient survival rates at 1 and 5 years for the MMF group were 97 and 94%, respectively, compared to 100% and 91% at 1 and 5 years respectively for the AZA group (P = 0.61). Graft survival rates at 1 and 5 years for the MMF group were 95 and 83%, respectively, compared to 97 and 84% at 1 and 5 years, respectively for the AZA group (P = 0.62). CONCLUSION: There was no difference in acute rejection episodes between MMF and AZA based immunotherapy. Additionally, we observed no significant difference concerning graft survival in the MMF group when compared to AZA group. PMID- 21622992 TI - Renal cell cancer--a multimodal approach to preserving renal function and improving survival. PMID- 21622993 TI - Comparable outcome of acute unplanned peritoneal dialysis and haemodialysis. AB - BACKGROUND: The impact of dialysis modality on outcome, especially on infection early in the course of dialysis, in unplanned acute dialysis initiation has not been well evaluated. The aim of the study was to compare the rates and causes of mortality and morbidity in incident dialysis patients started unplanned acute peritoneal dialysis (PD) or haemodialysis (HD). PATIENTS AND METHODS: In this observational cohort study, incident dialysis patients with initiation of unplanned and acute PD (n = 66) or HD (n = 57) at a single centre from March 2005 to June 2010 were included and followed up for 6 months (0-183 days, mean follow up time 4.72 months). For PD, surgically placed Tenckhoff catheters were used. All HD patients were dialysed with a central venous catheter (non-tunnelled or tunnelled). There were no significant differences in terms of gender, age and prevalence of diabetes mellitus in either group. The prevalence of heart failure [New York Heart Association (NYHA) Stage III-IV] was significantly higher in the PD group (73 versus 46% in HD group, P < 0.01). The population was stratified to PD and HD comparing mortality, infection, bacteraemia and hospitalization. RESULTS: Of the 123 patients who commenced acute and unplanned dialysis, n = 44 (35.8%) died during the follow-up period of 0-183 days. There were no significant difference in half-year mortality in n = 20 PD patients (30.3%) versus n = 24 HD patients (42.1%) (P = 0.19). The cardiovascular mortality in PD and HD patients were 9.1 and 10.5%, respectively (P = 1.00). Overall mortality due to infection was higher in the HD (17.5%) versus in the PD group (9.1%), however, not significant (P = 0.19). HD patients had significantly higher probability of bacteraemia in the first 183 days compared to PD patients (21.1 versus 3.0%, P < 0.01). Group comparison by Poisson regression analyses showed that the relative risk of bacteraemia in the PD group versus HD group was 0.16 (95% confidence interval, 0.05-0.57, P = 0.005). The significant difference was not affected by the confounder's patient age at time of dialysis, male sex, heart failure (NYHA III-IV), diabetes, malignancy and peripheral arterial occlusive disease Stage IV. There were high proportions of hospitalization after the initiation of dialysis in both groups (PD 75.0% and HD 67.3%, P = 0.40). Univariate and multiple regression analyses revealed only age at initiation of dialysis to be significantly associated with overall mortality (P < 0.05). CONCLUSIONS: Dialysis modality (PD versus HD) in an acute unplanned dialysis setting showed, in our population, no significant influence on survival. HD patients had a significantly higher risk of bacteraemia, perhaps due to central venous dialysis catheter. PD seems to be a safe and efficient, at least comparable, alternative to HD in acute unplanned dialysis settings. PMID- 21622994 TI - Uremia induces functional incompetence of bone marrow-derived stromal cells. AB - BACKGROUND: Chronic kidney disease (CKD) is associated with increased risk for cardiovascular diseases (CVD). We hypothesized that inadequate angiogenic response in uremic patients could result from dysfunction of bone marrow-derived stromal cells [mesenchymal stem cells (MSCs)]. METHODS: We investigated whether MSCs are functionally competent in uremia induced by partial kidney ablation in C57Bl/6J mice. RESULTS: Uremic MSCs showed decreased expression of vascular endothelial growth factor (VEGF), VEGF receptor (VEGFR)1 and stromal cell-derived factor (SDF)-1alpha, increased cellular senescence, decreased proliferation, defects in migration in response to VEGF and SDF-1alpha and in vitro tube formation. Interestingly, the expression of fibroblast-specific protein-1 was higher in uremic MSCs. Uremia decreased hypoxia-inducible factor-1alpha, VEGF and VEGFR1 expression under hypoxia and Akt phosphorylation in both basal and VEGF stimulated states. A diminished mitogenic effect on endothelial proliferation was observed in conditioned media from uremic MSCs. In addition, intravital microscopic analysis showed decreased angiogenesis in uremic MSCs. CONCLUSION: These results clearly demonstrate the functional incompetence in MSCs under uremic conditions and may significantly contribute to the disproportionately high risk for CVD in patients with CKD. PMID- 21622995 TI - A comprehensive study of polymorphic sites along the HLA-G gene: implication for gene regulation and evolution. AB - HLA-G molecule plays an important role on immune response regulation and has been implicated on the inhibition of T and natural killer cell cytolytic function and inhibition of allogeneic T-cell proliferation. Due to its immune-modulator properties, the HLA-G gene expression has been associated with the outcome of allograft and of autoimmune, infectious, and malignant disorders. Several lines of evidence indicate that HLA-G polymorphisms at the 5'-upstream regulatory region (5' URR) and 3'-untranslated region (3' UTR) may influence the HLA-G expression levels. Because Brazilians represent one of the most heterogeneous populations in the world with the widest HLA-G coding region variability already detected among the studied populations, a high level of variability and haplotype diversity would be expected in Brazilians. On this basis, the 5' URR, coding, and 3' UTR variability were evaluated in a Brazilian series consisting of 100 healthy bone marrow donors, as well as the linkage disequilibrium pattern along the gene and the extended haplotypes encompassing several gene segment variations. The HLA G locus seems to present six different HLA-G lineages showing functional variations mainly in nucleotides of the regulatory regions. Differences were observed at the 5' URR in positions that either coincide with or are close to transcription factor-binding sites and at the 3' UTR mainly in positions that have already been reported to influence HLA-G mRNA availability. We report several lines of evidence for balancing selection acting on the regulatory regions, which may indicate that these HLA-G lineages may be related to the differential HLA-G expression profiles. PMID- 21622996 TI - Bradycardia during critical care intubation: mechanisms, significance and atropine. AB - Bradycardia occurs during the intubation of some critically ill children as a result of vagal stimulation due to hypoxia and/or laryngeal stimulation; such 'stable' bradycardia is accompanied by selective vasoconstriction. Some induction drugs also induce bradycardia which may be accompanied by vasodilatation which is also a feature of certain pathologies, which influence the progression to 'unstable' bradycardia, which does not respond to re-oxygenation and a pause in laryngoscopy. Preintubation atropine diminishes the overall incidence of stable bradycardia during routine anaesthesia. However, clinical studies of critical care intubation show that atropine does not prevent all episodes of bradycardia and specifically cannot affect vasodilatation. As such, there is insufficient evidence to support a recommendation for the indiscriminate use of atropine for intubation during critical care illness, including simple neonatal respiratory distress. Atropine is appropriate during septic or late stage hypovolaemic shock where abnormal vasomotor tone and bradycardia may potentially set up a negative feedback loop of cardiac hypo-oxygenation and hypoperfusion and during premedication when using suxamethonium. PMID- 21622997 TI - Propranolol for infantile haemangiomas: a review. AB - Infantile haemangiomas are the most common benign tumour of infancy. However the majority are self-resolving and only a small minority of cases require treatment, with various different medications being used in the past. Over the last few years, propranolol, a non-selective beta-blocker, has become a popular and successful treatment for infantile haemangiomas. However, further research on its safety is needed if it is going to be used more frequently. This article summarises the current literature on propranolol for haemangioma treatment with emphasis on its toxicity and adverse event profile. PMID- 21622998 TI - Sleep duration and body mass index in 0-7-year olds. AB - OBJECTIVES: To determine in children aged 0-7 years (1) cross-sectional relationships between body mass index (BMI) and sleep duration, and whether (2) sleep duration predicts later BMI and/or (3) BMI predicts later sleep duration. DESIGN: Longitudinal Study of Australian Children, Waves 1 and 2. PARTICIPANTS: Infants aged 0-1 years (Wave 1), followed at age 2-3 years (Wave 2); children aged 4-5 years (Wave 1), followed at age 6-7 years (Wave 2). MAIN OUTCOME MEASURES: BMI, sleep duration by time-use diary, parent-reported sleep problems (none/mild vs moderate/severe). ANALYSES: Cross-sectional, ANOVA (sleep duration) and chi(2) (sleep problems); longitudinal, linear regression. RESULTS: 3857 (76%) infants and 3844 (77%) children had BMI and sleep data. At every wave, approximately 15% and 5% of children were overweight and obese, respectively. Obesity was not associated with sleep duration at 0-1, 2-3 or 4-5 years, though obese 6-7-year olds slept approximately 30 min less (p<0.001). Sleep problems were similar across BMI categories at all ages. Wave 1 sleep duration did not predict Wave 2 BMI, nor did Wave 1 BMI predict Wave 2 sleep duration. CONCLUSIONS: In these large child population cohorts, sleep duration did not predict obesity up to age 6-7 years. Current trials of sleep interventions to prevent or manage obesity in young children may be premature. PMID- 21622999 TI - Adiponectin and leptin increase IL-8 production in human chondrocytes. PMID- 21623000 TI - Aggressive combination drug therapy in very early polyarticular juvenile idiopathic arthritis (ACUTE-JIA): a multicentre randomised open-label clinical trial. AB - OBJECTIVES: In juvenile idiopathic arthritis (JIA), the efficacy of very early disease-modifying drug therapy, synthetic or biological, is not well known. Three alternative strategies were compared for treating recent-onset polyarticular JIA. METHODS: In a 54-week multicentre open-label clinical trial, 60 disease-modifying antirheumatic drug (DMARD)-naive patients aged 4-15 years were randomly assigned into three treatment arms. The efficacy of infliximab plus methotrexate (TNF) was compared to that of two synthetic therapies: methotrexate alone (MTX) and DMARD methotrexate, sulphasalazine and hydroxychloroquine in combination (COMBO). Primary endpoint was American College of Rheumatology paediatric 75% improvement (ACR Pedi 75). Secondary endpoints were inactive disease and safety. RESULTS: In 59 patients, mean (+/-SE) age at baseline was 9.6+/-0.4 years, duration of JIA 1.9+/-0.2 months and number of active joints 18+/-1. ACR Pedi 75 was achieved in 100% (19/19) of patients receiving TNF, 65% (13/20) on COMBO (95% CI 44% to 86%) and 50% (10/20) on methotrexate (95% CI 28% to 72%) p<0.0001. Thirteen patients receiving TNF (68%; 95% CI 47% to 89%) achieved inactive disease, whereas eight (40%; 95% CI 22% to 63%) on COMBO and five (25%; 95% CI 6% to 44%) on methotrexate did (p=0.002). Patients on TNF spent a mean 26 weeks (95% CI 18 to 34) with inactive disease, longer than did those receiving COMBO (13 weeks; 95% CI 6 to 20), or methotrexate (6 weeks; 95% CI 2 to 10). Serious adverse events were rare. CONCLUSION: In early polyarticular JIA, targeting to achieve minimally active or inactive disease, infliximab plus methotrexate was superior to synthetic DMARD in combination and strikingly superior to methotrexate alone. PMID- 21623001 TI - Assessment of dynamic humeral centering in shoulder pain with impingement syndrome: a randomised clinical trial. AB - OBJECTIVES: Treatment for degenerative rotator cuff disease of the shoulder includes physiotherapy. Dynamic humeral centering (DHC) aims at preventing subacromial impingement, which contributes to the disease. The goal of this study was to assess the effectiveness of DHC. METHOD: 69 patients with shoulder pain and impingement syndrome were prospectively included in a single-centre randomised trial with a 12-month follow-up. Patients and assessor were blinded to the study hypothesis and treatment, respectively. DHC and non-specific mobilisation as control were performed for 6 weeks, in 15 supervised individual outpatient sessions, and patients performed daily home exercises. The planned primary outcome was the Constant score including subscores for pain, activity, mobility and strength at 3 months. Secondary outcomes were the Constant score and subscores at 12 months, and medication use for pain at 3 and 12 months. RESULTS: The DHC group did not differ from the control group in the total Constant score at 3 months. However, the DHC group showed a higher Constant subscore for pain (12.2 (SD 2.8) vs 9.9 (2.9), least square means difference 2.1, 95% CI 0.7 to 3.5, p=0.004). At 3 months, the DHC group also showed a higher rate of no medication use (96.7% vs 71%, proportional difference 25.7, 95% CI 3.7 to 51.9, p=0.012). There was no other intergroup difference. CONCLUSIONS: There was no difference in the total Constant score between DHC and controls. However, pain was improved at 3 months after DHC. The differences found in subscores for pain should be explored in future studies. Trial registration clinicaltrials.gov Identifier: NCT 01022775. PMID- 21623002 TI - Skin fibroblasts are potent suppressors of inflammation in experimental arthritis. AB - OBJECTIVES: Mesenchymal stromal cells (MSC) are characterised by their capacity to suppress immune reactions. This function was reported to be shared in vitro by fibroblasts but their role has been poorly investigated in vivo. This study explored whether fibroblasts isolated from skin may suppress the host immune response in a model of autoimmune disorder. METHODS AND RESULTS: It was first confirmed that skin fibroblasts lack the capacity to differentiate into osteoblasts or chondrocytes but possess the capacity to inhibit in vitro the proliferation of T lymphocytes. Fibroblasts also secrete modulatory molecules, in particular prostaglandin E2 and nitric oxide, similar to MSC. To assess their role in vivo, the collagen-induced arthritis model was used, and showed that similar to MSC the intravenous injection of fibroblasts efficiently suppress clinical signs of arthritis and delay disease onset. This effect was associated with reduced inflammation as reflected by biological parameters and increased levels of IL-5, IL-10 and IL-13 in the spleens of treated mice. To characterise the mechanism of immunosuppression further, phenotypic analyses were performed and could not detect any induction of CD4 CD25 Foxp3(+) regulatory T (Treg) cells. A population of CD4 IL-10(+) T cells was, however, detected that was slightly increased after fibroblast injection and significantly upregulated after MSC administration. CONCLUSIONS: This study gives the first evidence for an immunosuppressive role of fibroblasts in vivo, and strongly suggests that fibroblasts induce a T-helper type 2 immune profile, although the possibility that IL-10-secreting Treg cells may be generated cannot be excluded. PMID- 21623003 TI - Confirmation of TNIP1 and IL23A as susceptibility loci for psoriatic arthritis. AB - OBJECTIVES: To investigate a shared genetic aetiology for skin involvement in psoriasis and psoriatic arthritis (PsA) by genotyping single-nucleotide polymorphisms (SNPs), reported to be associated in genome-wide association studies of psoriasis, in patients with PsA. METHODS: SNPs with reported evidence for association with psoriasis were genotyped in a PsA case and control collection from the UK and Ireland. Genotype and allele frequencies were compared between PsA cases and controls using the Armitage test for trend. RESULTS: Seven SNPs mapping to the IL1RN, TNIP1, TNFAIP3, TSC1, IL23A, SMARCA4 and RNF114 genes were successfully genotyped. The IL23A and TNIP1 genes showed convincing evidence for association (rs2066808, p = 9.1*10(-7); rs17728338, p = 3.5*10(-5), respectively) whilst SNPs mapping to the TNFAIP3, TSC1 and RNF114 genes showed nominal evidence for association (rs610604, p = 0.03; rs1076160, p = 0.03; rs495337, p = 0.0025). No evidence for association with IL1RN or SMARCA4 was found but the power to detect association was low. CONCLUSIONS: SNPs mapping to previously reported psoriasis loci show evidence for association to PSA, thus supporting the hypothesis that the genetic aetiology of skin involvement is the same in both PsA and psoriasis. PMID- 21623004 TI - Maternal diet, aging and diabetes meet at a chromatin loop. AB - We have recently demonstrated that exposure to a suboptimal diet during early development leads to abnormal epigenetic regulation of a promoter-enhancer interaction at the gene encoding HNF-4alpha, a key transcription factor required for pancreatic beta-cell differentiation and glucose homeostasis. In addition, our studies revealed that the suboptimal maternal diet amplifies the age associated epigenetic silencing of this locus. In this research perspective we discuss these novel findings in the context of the growing list of epigenetic mechanisms by which the environment can affect gene activity and emphasize their implications for the understanding of the mechanistic basis of the development of type 2 diabetes with age. PMID- 21623005 TI - Network modeling of MDM2 inhibitor-oxaliplatin combination reveals biological synergy in wt-p53 solid tumors. AB - Earlier we had shown that the MDM2 inhibitor (MI-219) belonging to the spiro oxindole family can synergistically enhance the efficacy of platinum chemotherapeutics leading to 50% tumor free survival in a genetically complex pancreatic ductaladenocarcinoma (PDAC) xenograft model. In this report, we have taken a systems and network modeling approach in order to understand central mechanisms behind MI219-oxaliplatin synergy with validation in PDAC, colon and breast cancer cell lines. Microarray profiling of drug treatments (MI-219, oxaliplatin or their combination) in capan-2 cells reveal a similar unique set of gene alterations that is duplicated in other solid tumor cells. As single agent, MI-219 or oxaliplatin induced alterations in 48 and 761 genes respectively. The combination treatment resulted in 767 gene alterations with emergence of 286 synergy unique genes. Ingenuity network modeling of combination and synergy unique genes showed the crucial role of five key local networks CREB, CARF, EGR1, NF-kB and E-Cadherin. Compared to single agents the combination treatment super induced p53 and p21 confirming functional synergy. Further, the network signatures were validated at the protein level in all three cell lines. Individually silencing central nodes in these five hubsinterfered with MI-219 oxaliplatin activity confirming their critical role in aiding p53 mediated apoptotic response. We anticipate that our MI219-oxaliplatin network blueprints can be clinically translated in the rationale design and application of this unique therapeutic combination in a genetically pre-defined subset of patients. PMID- 21623007 TI - High frequency stimulation abolishes thalamic network oscillations: an electrophysiological and computational analysis. AB - Deep brain stimulation (DBS) of the thalamus has been demonstrated to be effective for the treatment of epilepsy. To investigate the mechanism of action of thalamic DBS, we examined the effects of high frequency stimulation (HFS) on spindle oscillations in thalamic brain slices from ferrets. We recorded intracellular and extracellular electrophysiological activity in the nucleus reticularis thalami (nRt) and in thalamocortical relay (TC) neurons in the lateral geniculate nucleus, stimulated the slice using a concentric bipolar electrode, and recorded the level of glutamate within the slice. HFS (100 Hz) of TC neurons generated excitatory post-synaptic potentials, increased the number of action potentials in both TC and nRt neurons, reduced the input resistance, increased the extracellular glutamate concentration, and abolished spindle wave oscillations. HFS of the nRt also suppressed spindle oscillations. In both locations, HFS was associated with significant and persistent elevation in extracellular glutamate levels and suppressed spindle oscillations for many seconds after the cessation of stimulation. We simulated HFS within a computational model of the thalamic network, and HFS also disrupted spindle wave activity, but the suppression of spindle activity was short-lived. Simulated HFS disrupted spindle activity for prolonged periods of time only after glutamate release and glutamate-mediated activation of a hyperpolarization-activated current (I(h)) was incorporated into the model. Our results suggest that the mechanism of action of thalamic DBS as used in epilepsy may involve the prolonged release of glutamate, which in turn modulates specific ion channels such as I(h), decreases neuronal input resistance, and abolishes thalamic network oscillatory activity. PMID- 21623008 TI - Three-dimensional holographic photostimulation of the dendritic arbor. AB - Digital holography is an emerging technology that can generate complex light patterns for controlling the excitability of neurons and neural circuits. The strengths of this technique include a high efficiency with which available light can be effectively utilized and the ability to deliver highly focused light to multiple locations simultaneously. Here we demonstrate another strength of digital holography: the ability to generate instantaneous three-dimensional light patterns. This capability is demonstrated with the photolysis of caged glutamate on the dendritic arbor of hippocampal neurons, to study the nature of the integration of inputs arriving on multiple dendritic branches. PMID- 21623009 TI - Early detection of hand movements from electroencephalograms for stroke therapy applications. AB - Movement-assist devices such as neuromuscular stimulation systems can be used to generate movements in people with chronic hand paralysis due to stroke. If detectable, motor planning activity in the cortex could be used in real time to trigger a movement-assist device and restore a person's ability to perform many of the activities of daily living. Additionally, re-coupling motor planning in the cortex with assisted movement generation in the periphery may provide an even greater benefit-strengthening relevant synaptic connections over time to promote natural motor recovery. This study examined the potential for using electroencephalograms (EEGs) as a means of rapidly detecting the intent to open the hand during movement planning in individuals with moderate chronic hand paralysis following a subcortical ischemic stroke. On average, attempts to open the hand could be detected from EEGs approximately 100-500 ms prior to the first signs of movement onset. This earlier detection would minimize device activation delays and allow for tighter coupling between initial formation of the motor plan in the cortex and augmentation of that plan in the periphery by a movement-assist device. This tight temporal coupling may be important or even essential for strengthening synaptic connections and enhancing natural motor recovery. PMID- 21623010 TI - The central role of governance in community-based participatory research. PMID- 21623013 TI - Community-centered family health history: a customized approach to increased health communication and awareness. AB - BACKGROUND: There has been little study of whether family health history (FHH) tools used by individuals, families, and communities inspire measurable changes in communication and behavior. OBJECTIVES: The Community-Centered Family Health History (CCFHH) project was a collaborative endeavor among national and community based organizations with an interest in genetics education and health. Using community- based participatory research principles as a foundation, CCFHH examined whether the Does It Run In the Family? toolkit, a set of two customizable booklets on health and genetics, encourages discussion and collection of FHH information across diverse communities. METHODS: Five communities across the country measured the utility of customized versions of the Does It Run In the Family? toolkit. Each community partner recruited families, consisting of two or more blood relatives, to use the toolkit for 3 months, discuss it among their family members, and consider the implications of the health information. Pre- and postintervention surveys measured family communication about family history and disease risk and the use of FHH information in health care provider interactions. RESULTS: After aggregate, cross community analysis of individual responses, from pre- to post-toolkit use family members showed increases in communication about family history of disease risk (p < .05) and in awareness about FHH (p < .05). CONCLUSION: These findings indicate that diverse communities are receptive to FHH intervention, and tailored health educational materials can lead to increased conversations and awareness about health issues across communities. PMID- 21623014 TI - Closing the gap between insecticide treated net ownership and use for the prevention of malaria. AB - BACKGROUND: Malaria is the leading cause of morbidity and mortality in children younger than 5 years old and pregnant women in sub-Saharan Africa. Insecticide treated nets (ITNs) reduce clinical malaria by more than 50% and all cause mortality in young children by 15% to 30%. However, use of these nets is poor across sub-Saharan Africa, limiting the potential impact of this effective tool in the fight against malaria. OBJECTIVE: We sought to improve the use of ITNs using a community-created and -implemented approach, and measure the change in ITN use over the year after implementation. METHODS: Using a community-based participatory research approach, we created and implemented an intervention to improve ITN use in a rural village. Our intervention involved providing hands-on instructions and assistance in hanging of nets, in-home small group education, and monthly follow-up by trained community members. ITN use was measured for all individuals in a subset of the community (61 households, 759 individuals) at baseline and at 6 months and 1 year after distribution. RESULTS: Rates of individual usage increased significantly from 29% at baseline to 88.7% (p < .001) at 6 months and to 96.6% (p < .001) at 12 months. For children under age 5, usage rates increased from 46% at baseline to 95.7% (p < .001) at 6 months and 95.4% (p < .001) at 12 months. CONCLUSION: Our study demonstrates that rapidly achieving and sustaining almost universal ITN usage rates is possible using a community based approach. Closing the gap between ITN ownership and use will help communities to realize the full potential of ITNs in the prevention of malaria. PMID- 21623015 TI - Survivor voices: co-learning, re-connection, and healing through community action research and engagement (CARE). AB - This article describes the Trauma Healing Project (THP: www.http//healingattention.org), the Survivor Voices study (n = 351), and the complementary nature of community- campus partnerships (CCP) and community-based participatory action research methodology (PAR). Survivor Voices-a random digit telephone survey developed by, conducted, analyzed, and disseminated by survivors of abuse and violence, university researchers, and students-was designed to learn from survivors about what hurt and about what helped with regard to how people responded to their trauma, and what they recommend for trauma healing. We feature our CCP, including challenges faced, how we have addressed those challenges, and profile our current findings. We describe how PAR and CCPs can be very useful tools toward the development of a community-wide effort to reduce violence and support trauma healing. PMID- 21623016 TI - Collaboration strategies in nontraditional community-based participatory research partnerships: lessons from an academic-community partnership with autistic self advocates. AB - BACKGROUND: Most community-based participatory research (CBPR) projects involve local communities defined by race, ethnicity, geography, or occupation. Autistic self-advocates, a geographically dispersed community defined by disability, experience issues in research similar to those expressed by more traditional minorities. OBJECTIVES: We sought to build an academic-community partnership that uses CBPR to improve the lives of people on the autistic spectrum. METHODS: The Academic Autistic Spectrum Partnership in Research and Education (AASPIRE) includes representatives from academic, self-advocate, family, and professional communities. We are currently conducting several studies about the health care experiences and well-being of autistic adults. LESSONS LEARNED: We have learned a number of strategies that integrate technology and process to successfully equalize power and accommodate diverse communication and collaboration needs. CONCLUSIONS: CBPR can be conducted successfully with autistic self-advocates. Our strategies may be useful to other CBPR partnerships, especially ones that cannot meet in person or that include people with diverse communication needs. PMID- 21623017 TI - Promoting a bicycle-friendly environment: community assessment and action. AB - BACKGROUND: Researchers and community coalition members collected survey data and used it to guide production of an educational video promoting a bicycle-friendly environment. OBJECTIVES: Telephone and mailed surveys were conducted among bicyclists and motorists for needs assessment. METHODS: A sample of 173 bicyclists completed a mailed survey (response rate of 87.0%), and a sample of 801 motorists completed a telephone survey (response rate of 42.1%). Findings guided video production. RESULTS: Most motorists understood that bicyclists have a right to ride in the road, but about one half believed bicyclists create a hazard, even when they follow traffic laws. Bicyclists expressed safety concerns and felt threatened by motorists. Both motorists' and bicyclists' knowledge of traffic rules varied across the items. CONCLUSIONS: This project illustrates how community members can apply research findings to an educational video of high quality with local relevance. The leadership from community members in making the video set the foundation for local ownership. PMID- 21623018 TI - Developing a community action plan to eliminate cancer disparities: lessons learned. AB - BACKGROUND: African Americans bear an unequal burden of breast, cervical, and colorectal cancer. The Deep South Network for Cancer Control (DSN) is a community academic partnership operating in Alabama and Mississippi that was funded by the National Cancer Institute (NCI) to address cancer disparities using community based participatory research approaches. OBJECTIVE: In addition to reporting on the plans of this work in progress, we describe the participatory process that local residents and the DSN used to identify needs and priorities, and elaborate on lessons learned from applying a participatory approach to the development of a community action plan. METHODS: We conducted 24 community discussion groups involving health care professionals, government officials, faith-based leaders, and other stakeholders to identify cancer health disparity needs, community resources/assets, and county priorities to eliminate cancer health disparities. To develop a community action plan, four working groups explored the themes that emerged from the discussion groups, taking into consideration evidence-based strategies and promising community practices. RESULTS: The DSN formulated a community action plan focusing on (1) increasing physical activity by implementing a campaign for individual-level focused activity; (2) increasing the consumption of fruits and vegetables by implementing NCI's Body and Soul Program in local churches; (3) increasing cancer screening by raising awareness through individual, system, and provider agents of change; and (4) training community partners to become effective advocates. CONCLUSIONS: A community-academic partnership must involve trust, respect, and an appreciation of partners' strengths and differences. The DSN applied these guiding principles and learned pivotal lessons. PMID- 21623019 TI - Engaging nurse aide students to develop a survey to improve enrollment and retention in college. AB - BACKGROUND: Students from lower socioeconomic status (SES) backgrounds have historically experienced high rates of college dropout. Surveys often are used to assess supports and barriers (SB) to college enrollment and completion, and findings drive the design of interventions to improve student recruitment and retention. However, standard surveys may not include questions that solicit the breadth of issues facing low-income minority individuals. OBJECTIVES: We used community-based participatory research (CBPR) principles to develop an SB survey to better reflect the concerns of rural, first-generation college students in Hawai'i. METHODS: An advisory panel (AP) of students and community partners guided the work. The literature informed the first draft of the SB survey. Then we worked with students who had successfully completed a vocational Nurse Aide (NA) Training Program (NATP) course to refine four versions of the SB survey through multiple cycles of online survey review and focus groups. RESULTS: The final product included questions in new areas and differently phrased questions in standard areas (e.g., transportation, dependent care, housing, financial aid) to better capture reasons for students dropping out. The survey has proven useful as a student assessment tool, and findings are being used by instructors, counselors, and community partners to add resources and modify programs to increase student success in community college. CONCLUSIONS: Findings confirm the usefulness of engaging target partners in tool development. An enhanced understanding of SB of students from underrepresented groups will help to improve college recruitment and retention interventions. PMID- 21623020 TI - Meeting people where they are: engaging public housing residents for integrated pest management. AB - In a unique partnership, the Boston Public Health Commission, the Boston Housing Authority (BHA), Boston University School of Public Health, the Committee for Boston Public Housing, and the West Broadway Task Force (WBTF) led an Integrated Pest Management (IPM) intervention in Boston's public housing developments. Key to the success of the program was recruiting residents to participate. Residents who were trained as Community Health Advocates (CHAs) at the West Broadway Development in South Boston, Massachusetts, recruited over 300 homes to participate in the IPM intervention (out of 484 living units). This article describes the recruitment strategy and success from the perspective of CHAs at the West Broadway development. PMID- 21623021 TI - Development and evaluation of a toolkit to assess partnership readiness for community-based participatory research. AB - An earlier investigation by academic and community co-investigators led to the development of the Partnership Readiness for Community-Based Participatory Research (CBPR) Model, which defined major dimensions and key indicators of partnership readiness. As a next step in this process, we used qualitative methods, cognitive pretesting, and expert reviews to develop a working guide, or toolkit, based on the model for academic and community partners to assess and leverage their readiness for CBPR. The 75-page toolkit is designed as a qualitative assessment promoting equal voice and transparent, bi-directional discussions among all the partners. The toolkit is formatted to direct individual partner assessments, followed by team assessments, discussions, and action plans to optimize their goodness of fit, capacity, and operations to conduct CBPR. The toolkit has been piloted with two cohorts in the Medical University of South Carolina's (MUSC) Community Engaged Scholars (CES) Program with promising results from process and outcome evaluation data. PMID- 21623022 TI - CES4Health.info: an online tool for peer reviewed publication and dissemination of diverse products of community-engaged scholarship. AB - Community-engaged scholarship (CES)-research, teaching, programmatic and other scholarly activities conducted through partnerships between academic and community partners-may result in innovative applied products such as manuals, policy briefs, curricula, videos, toolkits, and websites. Without accepted mechanisms for peer-reviewed publication and dissemination, these products often do not "count" toward faculty promotion and tenure (P&T) and have limited opportunities for broad impact. This paper reports on CES4Health.info, a unique online tool for peer-reviewed publication and dissemination of products of CES in forms other than journal articles. In its first year, CES4Health.info has published 24 products and documented the satisfaction of users, authors, and reviewers. PMID- 21623024 TI - Connecting community with campus to address cancer health disparities: a community grants program model. AB - BACKGROUND: With growing interest in the CBPR approach to cancer health disparities research, mechanisms are needed to support adherence to its principles. The Carolina Community Network (CCN), 1 of 25 Community Network Programs funded by the National Cancer Institute (NCI), developed a model for providing funds to community-based organizations. OBJECTIVES: This paper presents the rationale and structure of a Community Grants Program (CGP) model, describes the steps taken to implement the program, and discusses the lessons learned and recommendations for using the grants model for CBPR. METHODS: Three types of projects-cancer education, implementation of an evidence-based intervention, and the development of community-academic research partnerships-could be supported by a community grant. The CGP consists of four phases: Pre-award, peer-review process, post-award, and project implementation. RESULTS: The CGP serves as a catalyst for developing and maintaining community-academic partnerships through its incorporation of CBPR principles. CONCLUSIONS: Providing small grants to community-based organizations can identify organizations to serve as community research partners, fostering the CBPR approach in the development of community academic partnerships by sharing resources and building capacity. PMID- 21623025 TI - World No Tobacco Day 2011: India's progress in implementing the framework convention on tobacco control. PMID- 21623026 TI - Superbug--the so-called NDM-1. PMID- 21623027 TI - Research on social inequalities in health in India. PMID- 21623028 TI - Endothelial nitric oxide synthase (eNOS) variants in cardiovascular disease: pharmacogenomic implications. PMID- 21623029 TI - Use of minocycline in viral infections. AB - Repurposing of old drugs is a useful concept as it helps to minimize costs associated with the research and development of a new drug. Minocycline, a common second generation antibiotic, has been shown to possess several other beneficial effects other than its intended uses. The antiviral role of minocycline has generated considerable interest from the last decade. It was first shown to be beneficial in preventing human immunodeficiency virus (HIV) infections and later it was reported to improve cognitive deficiencies associate with neuroAIDS. However, its antiviral efficacies are not limited to retroviruses alone. In animal models or in vitro systems of flaviviral infections (especially Japanese encephalitis virus), minocycline has been shown to be highly effective. However, not all effects are based on direct inhibition of viral replication. The general anti-inflammatory and immunomodulatory properties of minocycline are also responsible in part, in imparting the protective effects. Owing to the fact that minocycline is well tolerated by most people and that the drug has nearly 40 years history of usage, it is an exciting prospect to try out in other viral infections. PMID- 21623030 TI - Involvement of nitric oxide in learning & memory processes. AB - Nitric oxide (NO), synthesized from the amino acid, L-arginine by nitric oxide synthase (NOS) has received attention as a neurotransmitter in the brain. NO has been found to induce cognitive behaviour in experimental animals. In order to show evidence for the involvement of NO in learning and memory processes, the reports indicating the effects of its precursor, donors, and inhibitors of its synthesis in mammals, birds, fishes and invertebrates have been reviewed. Further, learning and memory impairment occurring in man and animals due to defective NO activity in the brain due to pathological conditions such as epilepsy, stress, diabetes and side effects of therapeutic agents and reversal of this condition by L-arginine and NO donors have been included. In addition, the reports that indicate ageing-induced impairment of cognition that is known to occur in Alzheimer's disease due to deposition of the toxic protein, beta amyloid and the effect of L-arginine and NO donors in preventing dementia in these patients have been reviewed. PMID- 21623031 TI - Socio-economic factors & longevity in a cohort of Kerala State, India. AB - BACKGROUND & OBJECTIVES: Even though Kerala State is well-known for its egalitarian policies in terms of healthcare, redistributive actions and social reforms, and its health indicators close to those of high-resource countries despite a poor per-capita income, it is not clear whether socio-economic disparities in terms of life expectancy are observed. This study was therefore carried out to study the impact of socio-economic level on life expectancy in individuals living in Kerala. METHODS: A cohort of 1,67,331 participants aged 34 years and above in Thiruvananthapuram district, having completed a lifestyle questionnaire at baseline in 1995, was followed up for mortality and cause of death until 2005. Survival estimates were based on the participants' vital status and death rates were calculated separately for men and women and for several socio-economic factors, stratified by age. RESULTS: At 40 years, men and women were expected to live another 34 and 37 years, respectively. Life expectancy varied across the participants' different socio-economic categories: those from high income households with good housing conditions, materially privileged households and small households, had a 2-3 years longer life expectancy as compared to the deprived persons. Also, those who went to college lived longer than the illiterates. The gaps between categories were wider in men than in women. INTERPRETATION & CONCLUSIONS: Socio-economic disparity in longevity was observed: wealthy people from Kerala State presented a longer life expectancy. PMID- 21623032 TI - Prevalence of endothelial nitric oxide synthase (eNOS) gene exon 7 Glu298Asp variant in North Eastern India. AB - BACKGROUND & OBJECTIVES: Endothelial nitric oxide is a potent vasodilator and impairment of its generation brought about by gene polymorphism is considered a major predictor for several diseases. A single nucleotide polymorphism G894T within exon 7 of endothelial nitric oxide synthase (eNOS-7) gene, resulting in a replacement of glutamic acid by aspartic acid, has been studied as a putative candidate gene for cardiovascular diseases. The pattern of eNOS-7 Glu298Asp variant in the Indian population is poorly known. The present study was planned to determine the prevalence of the variant of this gene among tea garden community in Assam, North-East India with high prevalence of hypertension. METHODS: Study participants of both sex aged >=18 yr were recruited randomly from temporary field clinics established in tea gardens of Dibrugarh, Assam. Genomic DNA was extracted from 409 subjects by the conventional phenol-chloroform method. The prevalence of the eNOS exon 7 Glu298Asp variant was determined by polymerase chain reaction and restriction fragment length polymorphism analysis. RESULTS: The study population was in Hardy-Weinberg Equilibrium. The frequency of the eNOS GG, GT and TT genotypes was found to be 75, 22 and 3 per cent respectively and did not show any significant difference in gender wise analysis. INTERPRETATION & CONCLUSIONS: Our results showed that the prevalence of the homozygous GG genotype was high (75%) and the rare mutant genotype (homozygous, TT) was 3 per cent in a population at risk with cardiovascular disease. Such population-based data on various polymorphisms can ultimately be exploited in pharmacogenomics. PMID- 21623033 TI - Normative data of cervical length in singleton pregnancy in women attending a tertiary care hospital in eastern India. AB - BACKGROUND & OBJECTIVES: The length of cervix predicts the risk of preterm delivery. The objective of this study was to assess cervical length in pregnancy by transvaginal ultrasonography for generating normative data for nulliparous women at no special risk of preterm labour. METHODS: An observational study was carried out in a tertiary care teaching hospital in eastern India in nulliparous women who delivered at term. A single sonologist assessed 224 women (once per subject) between 20 and 34 wk of gestation. Nulliparous women carrying a single foetus of confirmed gestational age were included; 216 subjects were finally considered for generation of normative data, excluding those delivering earlier than 37 or later than 42 wk. Other exclusion criteria were history of cerclage, any previous cervical surgery, smoking, or any medical disorder complicating pregnancy. RESULTS: Cervix length at each week of gestation gradually decreased over the study period. Length at 20 and 34 wk was 40.5 +/- 1.14 mm (mean +/- SD) and 34.8 +/- 1.34 mm respectively. The overall shortening over this 14 wk period was 5.7 mm, with 0.58 mm per week median rate of shortening. Pearson's correlation coefficient was -0.69 (95% CI -0.75 to -0.60; P< 0.001) for cervical length vis-a-vis gestational age. INTERPRETATION & CONCLUSIONS: The serial normative data generated in our setting can be used to decide cut-off points for predicting risk of preterm labour in future studies. Validity of such prediction needs to be tested in larger cohorts of women assessed at specific gestational ages. PMID- 21623034 TI - Study on cow ghee versus soybean oil on 7,12-dimethylbenz(a)-anthracene induced mammary carcinogenesis & expression of cyclooxygenase-2 & peroxisome proliferators activated receptor-gamma in rats. AB - BACKGROUND & OBJECTIVES: Breast cancer is a leading cause of cancer death in women; dietary fat is the one of the factors that influences its incidence. In the present study we investigated the effect of feeding cow ghee versus soybean oil on 7,12-dimethylbenz(a)anthracene (DMBA) induced mammary cancer in rat and expression of cyclooxygenase-2 and peroxisome proliferators activated receptor gamma (PPAR-gamma) in mammary gland. METHODS: Two groups of 21 day old female rats (30 each) were fed for 44 wk diet containing cow ghee or soybean oil (10%). The animals were given DMBA (30 mg/kg body weight) through oral intubation after 5 wk feeding. Another two groups (8 each) fed similarly but not given DMBA served as control for the gene expression study. RESULTS: In DMBA treated groups, the animal fed soybean oil had higher tumour incidence (65.4%), tumour weight (6.18 g) and tumour volume (6285 mm3) compared to those fed cow ghee (26.6%, 1.67 g, 1925 mm3, respectively). Tumour latency period was 23 wk on soybean oil compared to 27 wk on cow ghee. Histological analysis of tumours showed that the progression of carcinogenesis was more rapid on soybean oil than on cow ghee. The expression of cyclooxygenase-2 was observed only in DMBA treated rats and it was significantly less on cow ghee than on soybean oil. The expression of PPAR-gamma was significantly more on cow ghee than on soybean oil. INTERPRETATION & CONCLUSIONS: Our results show that dietary cow ghee opposed to soybean oil attenuates mammary carcinogenesis induced by DMBA; and the effect is mediated by decreased expression of cyclooxygenase-2 and increased expression of PPAR-gamma in the former group. PMID- 21623035 TI - Fibrous dysplasia & McCune-Albright syndrome: an experience from a tertiary care centre in north India. AB - BACKGROUND & OBJECTIVES: Fibrous dysplasia (FD) is a rare metabolic bone disease and information available from India is limited to only anecdotal case reports. We describe the clinical profile and therapeutic outcome of 25 patients with FD observed over a period of 14 yr in a tertiary care centre from north India. METHODS: In this retrospective study patients (n = 25) with diagnosis of fibrous dysplasia based on either classical radiological features and/or histological evidence on bone biopsy, were analyzed. Associated endocrinopathies if any, were evaluated. The diagnosis of McCune Albright syndrome (MAS) was considered when fibrous dysplasia was accompanied by either cafe-au-lait macules and/or endocrinopathies. The clinical presentation, biochemical parameters and imaging were analysed. Seven patients received bisphosphonate therapy. The final outcome and side effects were noted. RESULTS: Age of the patients ranged from 7 to 48 yr (mean +/- SD, 24.2 +/- 11.4 yr) with a lag time between onset of symptoms and presentation ranging from 1 to 20 yr (mean +/- SD, 6.6 +/- 6.2 yr). The mean duration of follow up was 3.5 +/- 2.1 yr. Eighteen (72%) patients had polyostotic disease while the remaining had monostotic FD. Eight patients had endocrinopathies: five had acromegaly, one each had gonadotropin independent precocious puberty (GIPP), hyperthyroidism and hypophosphatemic rickets. One child with GIPP later developed hyperthyroidism. McCune Albright syndrome was observed in 10 (40%) patients. A majority of the patients underwent various minor or major surgical procedures and seven patients received bisphosphonates for recurrent pathological fractures. Bone pain was reduced in all bisphosphonate treated patients with a decrease in subsequent fractures. INTERPRETATION & CONCLUSIONS: This series of FD patients from north India shows the varying presentations of this rare disease. Medical treatment with bisphosphonates appears to be potentially rewarding. PMID- 21623036 TI - Profile of bacterial pneumonia during Hajj. AB - BACKGROUND & OBJECTIVES: The congregation of a large number of people during Hajj seasons from different parts of the world in overcrowded conditions within a confined area for a long period of time presents many public health challenges and health risks. One of the main health problems of the crowding is ease transmission of pneumonia by air droplets. This study was aimed to determine the most common causes of bacterial pneumonia during the 2005 Hajj season and to relate the findings with clinical conditions. METHODS: A total of 141 patients with suspected pneumonia from the three main tertiary care hospitals in Makkah, Saudi Arabia, were investigated during Hajj season, 2005. Sputum and serum samples were collected and investigated for the possible presence of typical or atypical causative agents. RESULTS: Of the 141 clinically suspected pneumonia cases, 76 (53.9%) were confirmed positive by microbiological tests. More than 94 per cent of the confirmed cases were in the age group >50 yr, and 56.6 per cent of the cases were men. The most frequent isolates were Candida albicans (28.7%) and Pseudomonas aeruginosa (21.8%), followed by Legionella pneumophila (14.9%) and Klabsiella pneumoniae (9.2%). More than one causative pathogens were isolated in 15 patients (16.3%), and 55 per cent of patients were diabetic. INTERPRETATION & CONCLUSIONS: Clinicians should be aware that typical pneumonia treatment regimens may not work well during the Hajj season due to the wide variety of isolated organisms. This necessitates taking a sputum sample before starting treatment for identification and sensitivity testing. Special precautions need to be taken for >50 yr old patients. PMID- 21623037 TI - Virulence attributes of Helicobacter pylori isolates & their association with gastroduodenal disease. AB - BACKGROUND & OBJECTIVES: Certain genotype(s) of Helicobacter pylori strains may play important role in the development of gastric cancer (GC) and peptic ulcer disease (PUD). This study was undertaken to investigate the association of cagA, cagA3/ region subtypes, babA2 and vacA genotypes of H. pylori with GC, PUD and non-ulcer dyspepsia (NUD) as there are no such studies from India. METHODS: A total of 348 consecutive adult patients (NUD 241, PUD 45, GC 62) undergoing upper gastrointestinal endoscopy between September 2002 and May 2007 in a tertiary referral centre at Lucknow, north India, were enrolled. H. pylori infection was diagnosed by rapid urease test, culture, histopathology and PCR. Genotyping for cagA, cagA3/ subtypes, babA2 and vacA was performed by PCR using sequence specific primers. RESULTS: H. pylori infection was higher in patients with PUD than with GC (80 vs. 56.5%, P < 0.01) and NUD (80 vs. 55.2%, P= 0.002). cagA positive H. pylori isolates were detected in 80 per cent in GC, 83.3 per cent in PUD and 76.7 per cent in NUD with no significant difference among them. Only A subtype of cagA3/ was detected and its distribution in GC, PUD and NUD was 68.8, 69.4 and 52.6 per cent respectively. Presence of babA2 genotype was 31.4 per cent and it had significant association with PUD when compared with NUD (52.8 vs. 26.3%, P<0.003). On univariate regression analysis, s1a allele was associated with GC (P<0.050) and s1a/m2 vacA genotype with both GC (P=0.014) and PUD (P=0.016). INTERPRETATION & CONCLUSIONS: H. pylori infection was strongly associated with PUD with a very high proportion of patients with GC have s1a allele and s1a/m2 vacA genotype. Both s1a/m2 vacA genotype and babA2 are associated with PUD. The study shows that different virulence attributes of H. pylori are involved in different gastroduodenal disorders. PMID- 21623038 TI - Clinical response of newly diagnosed HIV seropositive & seronegative pulmonary tuberculosis patients with the RNTCP Short Course regimen in Pune, India. AB - BACKGROUND & OBJECTIVES: In the Revised National Tuberculosis Control Programme (RNTCP) in India prior to 2005, TB patients were offered standard DOTS regimens without knowledge of HIV status. Consequently such patients did not receive anti retroviral therapy (ART) and the influence of concomitant HIV infection on the outcome of anti-tuberculosis treatment remained undetermined. This study was conducted to determine the results of treatment of HIV seropositive pulmonary tuberculosis patients with the RNTCP (DOTS) regimens under the programme in comparison with HIV negative patients prior to the availability of free ART in India. METHODS: Between September 2000 and July 2006, 283 newly diagnosed pulmonary TB patients were enrolled in the study at the TB Outpatient Department at the Talera Hospital in the Pimpri Chinchwad Municipal Corporation area at Pune (Maharashtra): they included 121 HIV seropositive and 162 HIV seronegative patients. They were treated for tuberculosis as per the RNTCP in India. This study was predominantly conducted in the period before the free ART become available in Pune. RESULTS: At the end of 6 months of anti-TB treatment, 62 per cent of the HIV seropositive and 92 per cent of the HIV negative smear negative patients completed treatment and were asymptomatic; among smear positive patients, 70 per cent of the HIV-seropositive and 81 per cent of HIV seronegative pulmonary TB patients were cured. Considering the results in the smear positive and smear negative cases together, treatment success rates were substantially lower in HIV positive patients than in HIV negative patients, (66% vs 85%). Further, 29 per cent of HIV seropositive and 1 per cent of the HIV seronegative patients expired during treatment. During the entire period of 30 months, including 6 months of treatment and 24 months of follow up, 61 (51%) of 121 HIV positive patients died; correspondingly there were 6 (4%) deaths among HIV negative patients. INTERPRETATION & CONCLUSIONS: The HIV seropositive TB patients responded poorly to the RNTCP regimens as evidenced by lower success rates with chemotherapy and high mortality rates during treatment and follow up. There is a need to streamline the identification and management of HIV associated TB patients in the programme with provision of ART to achieve high cure rates for TB, reducing mortality rates and ensuring a better quality of life. PMID- 21623039 TI - Outcome of standardized treatment for patients with MDR-TB from Tamil Nadu, India. AB - BACKGROUND & OBJECTIVES: Programmatic management of MDR-TB using a standardized treatment regimen (STR) is being implemented under the Revised National Tuberculosis Control Programme (RNTCP) in India. This study was undertaken to analyse the outcomes of MDR-TB patients treated at the Tuberculosis Research Centre, Chennai, with the RNTCP recommended 24 months STR, under programmatic conditions. METHODS: Patients failed to the category II re-treatment regimen and confirmed to have MDR-TB, were treated with the RNTCP's STR in a prospective field trial on a predominantly ambulatory basis. Thirty eight patients were enrolled to the trial from June 2006 to September 2007. RESULTS: Time to culture conversion was two months or less for 82 per cent of patients. Culture conversion rates at 3 and 6 months were 84 and 87 per cent respectively. At the end of treatment, 25 (66%) were cured, 5 defaulted, 3 died and 5 failed. At 24 months, 30 (79%) patients, including 5 defaulters, remained culture negative for more than 18 months. Twenty two (58%) patients reported adverse drug reactions (ADRs) which required dose reduction or termination of the offending drug. No patient had XDR-TB initially, but 2 failure cases emerged as XDR-TB during treatment. INTERPRETATION & CONCLUSIONS: Outcomes of this small group of MDR-TB patients treated with the RNTCP's STR is encouraging in this setting. Close attention needs to be paid to ensure adherence, and to the timely recognition and treatment of ADRs. PMID- 21623040 TI - Effect of efflux pump inhibitors on drug susceptibility of ofloxacin resistant Mycobacterium tuberculosis isolates. AB - BACKGROUND & OBJECTIVES: In drug resistant, especially multi-drug resistant (MDR) tuberculosis, fluoroquinolones (FQs) are used as second line drugs. However, the incidence of FQ-resistant Mycobacterium tuberculosis is rapidly increasing which may be due to extensive use of FQs in the treatment of various other diseases. The most important known mechanism i.e., gyrA mutation in FQ resistance is not observed in a significant proportion of FQ resistant M. tuberculosis isolates suggesting that the resistance may be because of other mechanisms such as an active drug efflux pump. In this study we evaluated the role of the efflux pumps in quinolone resistance by using various inhibitors such as carbonyl cyanide m chlorophenyl hydrazone (CCCP), 2,4-dinitrophenol (DNP) and verapamil, in clinical isolates of M. tuberculosis. METHODS: A total of 55 M. tuberculosis clinical isolates [45 ofloxacin (OFL) resistant and 10 ofloxacin sensitive] were tested by Resazurin microtitre assay (REMA) to observe the changes in ofloxacin minimum inhibitory concentration (MIC) levels in presence of efflux inhibitors as compared to control (without efflux inhibitor). RESULTS: The MIC levels of OFL showed 2-8 folds reduction in presence of CCCP (16/45; 35.5%), verapamil (24/45; 53.3%) and DNP (21/45; 46.6%) while in case of isolates identified as OFL sensitive these did not show any effect on ofloxacin MICs. In 11 of 45 (24.5%) isolates change in MIC levels was observed with all the three inhibitors. Overall 30 (66.6%) isolates had reduction in OFL MIC after treatment with these inhibitors. A total of eight isolates were sequenced for gyrA gene, of which, seven (87.5%) showed known mutations. Of the eight sequenced isolates, seven (87.5%) showed 2 to 8 fold change in MIC in presence of efflux inhibitors. INTERPRETATION & CONCLUSIONS: Our findings suggest the involvement of active efflux pumps of both Major Facilitator Super Family (MFS) family (inhibited by CCCP and DNP) and ATP Binding Cassette (ABC) transporters (inhibited by verapamil) in the development of OFL resistance in M. tuberculosis isolates. Epidemiological significance of these findings needs to be determined in prospective studies with appropriate number of samples/isolates. PMID- 21623041 TI - Detection & characterization of Shiga toxin producing Escherichia coli (STEC) & enteropathogenic Escherichia coli (EPEC) in poultry birds with diarrhoea. AB - BACKGROUND & OBJECTIVES: Limited information is available on shiga toxin producing Escherichia coli (STEC) in animals and birds from India. An outbreak of acute diarrhoea in poultry birds at Aizawl, Mizoram was investigated for detection and characterization of STEC and enteropathogenic E. coli (EPEC). METHODS: E. coli was isolated and identified from rectal swabs, intestinal contents, heart blood and spleen of 19 poultry birds that died due to acute diarrhoea during the outbreak. Phenotypic characterization was done by standard bacteriological and biochemical techniques. All the isolates were serotyped based on their somatic antigens. Virulence genes (stx 1, stx 2, eaeA and hlyA) were detected by multiplex PCR assay. RESULTS: A total of 42 E. coli isolates were obtained, of which 24 belonged to 3 serogroups (O64, O89 and O91) and the remaining 18 were untypable (UT). Altogether, 14 (33.33%) isolates carried at least 1 virulence gene, of which 10 (23.81%) and 4 (9.52%) were recorded as STEC and EPEC, respectively. Of the 10 STEC isolates, one carried only stx2 , one carried stx 2 and hlyA, four carried stx1 , stx2 and hlyA, two carried stx 1, eaeA and hlyA genes and two carried stx 1 and eaeA. Of the four EPEC isolates, two carried eaeA and hlyA, one carried only eaeA gene and 1 carried only hlyA gene. INTERPRETATION & CONCLUSIONS: This is the first report on the involvement of STEC in poultry in India. PMID- 21623042 TI - Treatment practices & laboratory investigations during chikungunya outbreaks in South India. PMID- 21623043 TI - Phylogenetic analysis of swine & human hepatitis E virus. PMID- 21623044 TI - Sperm DNA damage & oxidative stress in recurrent spontaneous abortion (RSA). PMID- 21623045 TI - Are women with mental illness & the mentally challenged adequately protected in India? PMID- 21623046 TI - Prevalence and determinants of depression in type 2 diabetes patients: problem in generalization. PMID- 21623047 TI - Need for better anthropometric markers for prediction of cardiovascular risk in nutritionally stunted populations. PMID- 21623048 TI - Introducing pentavalent vaccine in EPI in India: a counsel for prudence in interpreting scientific literature. PMID- 21623049 TI - Laboratory quality control based on risk management. AB - Risk management is the systematic application of management policies, procedures, and practices to the tasks of analyzing, evaluating, controlling and monitoring risk (the effect of uncertainty on objectives). Clinical laboratories conduct a number of activities that could be considered risk management including verification of performance of new tests, troubleshooting instrument problems and responding to physician complaints. Development of a quality control plan for a laboratory test requires a process map of the testing process with consideration for weak steps in the preanalytic, analytic and postanalytic phases of testing where there is an increased probability of errors. Control processes that either prevent or improve the detection of errors can be implemented at these weak points in the testing process to enhance the overall quality of the test result. This manuscript is based on a presentation at the 2nd International Symposium on Point of Care Testing held at King Faisal Specialist Hospital in Riyadh, Saudi Arabia on October 12-13, 2010. Risk management principles will be reviewed and progress towards adopting a new Clinical and Laboratory Standards Institute Guideline for developing laboratory quality control plans based on risk management will be discussed. PMID- 21623050 TI - Six-year outcome of the national premarital screening and genetic counseling program for sickle cell disease and beta-thalassemia in Saudi Arabia. AB - BACKGROUND AND OBJECTIVES: Saudi Arabia has a high prevalence of hereditary hemoglobin disorders. Data has been collected by the Saudi Premarital Screening and Genetic Counseling Program on the prevalence of sickle cell disease and beta thalassemia but the outcomes were not quantified. We used six years of premarital screening data to estimate the burden of sickle disease and beta-thalassemia over the program period and to assess the frequency of at-risk marriage detection and prevention. DESIGN AND SETTING: Retrospective review, premarital couples attending premarital and genetic counseling clinics with marriage proposals between 2004 and 2009. METHODS: Blood samples obtained from all couples with marriage proposals between 2004 and 2009 were tested for sickle cell disease and beta-thalassemia. Test results were shared with all examinees and genetic counseling was offered for all at-risk couples. Marriage certificates were issued irrespective of the results and compliance with medical advice was voluntary. RESULTS: Out of all men and women examined, 70,962 (4.5%) and 29,006 (1.8%) were carriers or cases of sickle cell disease and beta-thalassemia, respectively. While the prevalence of sickle cell disease was constant between 2004 and 2009 (average 45.1 per 1000 examined persons, P=.803), the prevalence of beta thalassemia steadily decreased from 32.9 to 9.0 per 1000 examined persons (P<.001). The frequency of at-risk couples decreased by about 60% between 2004 and 2009 (from 10.1 to 4.0 per 1000 examined persons, P<.001). The frequency of voluntary cancellation of marriage proposals among at-risk couples showed more than 5-fold increase between 2004 and 2009 (from 9.2% to 51.9%, P<.001). The eastern region had 58% of all detected at-risk marriages and showed the greatest decline in detection and increase in prevention over time compared to other regions of Saudi Arabia. CONCLUSION: Six years of premarital screening in Saudi Arabia markedly reduced the number of at-risk marriages, which may considerably reduce the genetic disease burden in Saudi Arabia in the next decades. PMID- 21623051 TI - Factors affecting the progression of diabetic nephropathy and its complications: a single-center experience in Saudi Arabia. AB - BACKGROUND AND OBJECTIVES: One out of five Saudi diabetics develops end-stage renal disease (ESRD). Factors associated with progressive loss of renal function have not been extensively studied and reported in our community. We sought to evaluate the pattern and progression in glomerular filtration rate (GFR) and investigate the potential risk factors associated with progression to diabetic nephropathy (DN) among Saudi patients. DESIGN AND SETTING: Hospital-based retrospective analysis of type 2 diabetic patients seen between January 1989 and January 2004 at Security Forces Hospital and King Saud University in Riyadh, Saudi Arabia. PATIENTS AND METHODS: DN was defined as persistent proteinuria assessed by urine dipstick [at least twice for at least two consecutive years and/or serum creatinine >130 MUmol/L; and/or GFR <60 mL/min/1.73m(2) ]. RESULTS: Of 1952 files reviewed, 621 (31.8%) met the criteria for DN, and 294 (47%) were males. The mean (SD) age of the patients at baseline was 66.9 (11.4) years, and mean duration of diabetes was 15.4 (7.5) years. GFR deteriorated from a baseline value of 78.3 (30.3) mL/min/1.73m(2) to 45.1 (24.1) mL/min/1.73m(2) at the last visit, with a mean rate of decline in GFR of 3.3 mL/min/year. Progression of nephropathy was observed in 455 (73.3%) patients, with 250 (40.3%) patients doubling their first-hospital-visit serum creatinine level in a mean of 10.0 (6.0) years. At the end of the study, 16.5% of the cohort developed ESRD and were dialyzed. GFR >90 mL/min/1.73m(2) at the first hospital visit; duration of diabetes >10 years; persistent proteinuria; systolic blood pressure >130 mm Hg; and presence of retinopathy were significant markers associated with progression of nephropathy. CONCLUSION: Diabetic nephropathy tends to be progressive among Saudis, with GFR deteriorating at a rate of 3.3 mL/year and with a doubling of serum creatinine level in 40.3% of patients in 9.9 years. PMID- 21623052 TI - Pattern of presentation in type 1 diabetic patients at the diabetes center of a university hospital. AB - BACKGROUND AND OBJECTIVES: Diabetes mellitus (DM) is a major health problem worldwide. This study aimed to investigate the pattern of presentation and complications of pediatric diabetes. DESIGN AND SETTING: Retrospective study of children treated at a diabetes clinic at a university hospital for diabetes over 12-year period. PATIENTS AND METHODS: We collected data on the age at onset, sex, clinical presentation, duration of symptoms before diagnosis, and partial remission rate that were obtained from the hospital medical records, the National Diabetes Registry, and the statistics department. RESULTS: Of 369 diabetic children, most (n=321) children had polyuria (92%) 321/369=87% as the presenting symptom; other symptoms included polydipsia (310 patients, 88.8% 310/369=84%), weight loss (292 patients, 83.9%), nocturia (240 patients, 68.8% 240/369=65%), diabetic ketoacidosis (DKA) (174 patients, 49.9% 174/369=47.20%), and abdominal pain (172 patients, 49.3% 174/369=46.6%). Presenting symptoms were missing in 20 files, so the percentages were calculated among 349 patients. Most patients had acute diabetic complications such as hypoglycemia (222 patients, 62%) and DKA (88 patients, 38.1%, but none had severe complications such as coma and cerebral edema. Chronic complications included retinopathy (4 patients, 1.3%), neuropathy (2 patients, 0.6%), coronary heart disease (2 patients, 0.6%), and nephropathy (1 patient, 0.4%). CONCLUSION: The pattern of presentation of type 1 diabetes has changed as the incidence of DKA has decreased; unlike in previous studies, DKA was not the most common presenting symptom in this study. Chronic complications of diabetes, such as retinopathy, neuropathy, coronary heart disease, and nephropathy are mostly rare but still present. These complications might be prevented by achieving better awareness of the need for glycemic control. PMID- 21623053 TI - Clinical and autonomic functions: A study of childhood anxiety disorders. AB - BACKGROUND AND OBJECTIVES: Childhood and adolescent anxiety is generally associated with a varied somatic symptom pattern thought to reflect autonomic system activity. Few studies have examined the autonomic characteristics of generalized anxiety disorder (GAD). This omission is at odds with contemporary models of autonomic cardiovascular control. The current study aimed to find differences in autonomic functions between children with a diagnosis of childhood anxiety disorder and a control group using a case-control design. DESIGN AND SETTING: A cross sectional experimental study conducted in the years 2004-2005 in the psycho-physiology lab of a tertiary care multi-speciality teaching hospital. METHODS: Assessments were carried out using a semistructured interview, K-SADS (Schedule for Affective Disorders and Schizophrenia for Children and Adolescents); STAIC (State and Trait Anxiety Inventory for Children); CDRS (Childhood Depression Rating Scales); SCARED (Self-Report for Childhood Anxiety Related Disorders). Autonomic reactivity was tested using the standard battery of tests. RESULTS: There were differences between 34 children and adolescents (age range, 8-18 years) with a diagnosis of childhood anxiety disorder and a control group of 30 age- and sex-matched subjects from a nearby school in autonomic activity and reactivity between individuals with anxiety disorder and non-anxious control subjects. Our finding is suggestive of autonomic rigidity or diminished physiologic flexibility in children with anxiety disorder. CONCLUSIONS: The study is probably the first of its kind to look into the issue in detail using a detailed battery of the autonomic function tests, and the results are of help in better understanding the condition. The result of the present experiment supports differences in autonomic activity and reactivity between individuals with anxiety disorder and non-anxious control subjects. PMID- 21623054 TI - Antioxidant defense and oxidative stress in children with acute hepatitis A. AB - BACKGROUND AND OBJECTIVES: Published data on oxidative stress in children with acute hepatitis A are still very scarce. This study aims to evaluate the oxidant/antioxidant status of these patients. DESIGN AND SETTING: Prospective, case-control study, over 2.5 years in patients under hospitalized and ambulatory care. PATIENTS AND METHODS: The levels of a whole-blood antioxidant, reduced glutathione; and plasma antioxidants, beta-carotene, retinol, ascorbic acid, alpha-tocopherol; and the biomarker of oxidative stress, malondialdehyde, were evaluated in 50 pediatric patients (age range, 5-16 years; 29 males and 21 females) with acute hepatitis A and in 50 healthy children as control subjects (age range, 5-16 years; 25 males and 25 females). RESULTS: Plasma levels of reduced glutathione, beta-carotene, retinol, alpha-tocopherol and ascorbic acid were significantly lower, while malondialdehyde plasma levels were significantly increased in the patient group when compared to the controls (P<.0001 for all parameters). CONCLUSIONS: Our findings show that pediatric patients with acute hepatitis A were influenced by oxidative stress, resulting in significantly lower levels of plasma antioxidants and increased lipid peroxidation. In the absence of other therapeutic options, antioxidant vitamin supplements could be added to the therapy for these patients to help reestablish the oxidant status balance. Further investigations to confirm this suggestion are recommended. PMID- 21623055 TI - Sexually transmitted infections in Pakistan. AB - BACKGROUND AND OBJECTIVES: Sexually transmitted infections (STIs) represent a major global health problem leading to morbidity, mortality and stigma. Prior to this study there was no information on the prevalence and knowledge of STIs in Faisalabad, Pakistan. DESIGN AND SETTING: Prospective, cross-sectional study in patients attending STI clinics from July 2006 to September 2009. PATIENTS AND METHODS: After obtaining consent, patients completed structured questionnaires used for behavioral surveys. Blood and urethral swabs were collected and tested for syphilis, gonococcus, genital herpes, chlamydia and chancroid. RESULT: Mean (standard deviation) age of the 1532 participants was 38.9 (9.4) years, including 37.8 (10.2) years for males and 35.5 (6.3) years females. Male gender (n=1276, 83.3%), low socioeconomic class (n=1026, 67.0%) and residence in rural suburbs (n=970, 63.3%) were more common. Most (n=913, 59.6%) were aware of the modes of transmission of STIs and the associated complications, 20% (n=306) were condom users, and 21.2% (n=324) had knowledge of safe sex. Opposite-sex partners were preferred by 972 (63.4%) patients, while 29.9% (n=458) had both homosexual and heterosexual sex partners. Syphilis was present in 29.5% of patients (n=452); gonorrhea, in 13% (n=200), HSV-2, in 3.2% (n=49), chlamydia, in 4.7% (n=72) and chancroid, in 1.3% (n=20). CONCLUSION: This report establishes baseline local prevalence rates for STIs. Syphilis emerged as the most prevalent STI in Faisalabad. Population-based studies are required to study the epidemiology of STIs, along with initiation of national health-education campaign. PMID- 21623056 TI - Epidemiological and clinical features of Brucella arthritis in 24 children. AB - BACKGROUND AND OBJECTIVES: Brucellosis is considered the leading zoonotic disease of the Middle East. The disease has a wide spectrum of clinical manifestations and can result in complications with severe morbidity. The aim of this study was to evaluate the frequency, distribution and characteristics of arthritis in Iranian children with brucellosis. DESIGN AND SETTING: A retrospective descriptive study conducted in a referral children center in Tehran from 1997 to 2005. PATIENTS AND METHODS: Arthritis in children with brucellosis who were admitted to a specialized hospital was detected by clinical signs of the involved joint(s) and characteristics of joint fluid aspiration. Socio-demographic information was recorded. RESULTS: Of 96 patients diagnosed with brucellosis, 24 (25%) had Brucella arthritis 14 (58.3%) males and 10 (41.7%) females. Most common manifestations were fever in 21 patients (87.5%) and fatigue in 18 patients (75%). Monoarthritis was recorded in 15 patients (62.5%) of the cases with involvement of the knee in 8 (45%) and hip in 5 (29%), the ankle in 2 (8%) patients while 9 (37.5%) patients suffered from polyarthritis. None of the patients had axial joints involvement. Seventy-five percent of the subjects (18 patients) were from urban areas and 66.7% (16 patients) had consumed un pasteurized cheese. Recurrence was not seen in any of the 24 patients who received a combination of co-trimoxazole for 6 weeks and gentamicin for 5 days. CONCLUSION: Childhood brucellosis is a challenging disease in Iran that has serious complications like arthritis. Therefore all physicians who work in endemic areas should be familiar with this disease and consider the possibility of brucellosis in all children who present with arthritis and arthralgia. PMID- 21623057 TI - Incidental parathyroidectomy during thyroid resection: incidence, risk factors, and outcome. AB - BACKGROUND AND OBJECTIVES: Thyroidectomy is a commonly performed procedure for thyroid problems. Inadvertent removal of the parathyroid glands is one of its recognized complications, which occurs more frequently in certain high-risk patients. The aim of this study was to identify the incidence, risk factors, and clinical relevance of incidental parathyroidectomy during thyroid surgery. DESIGN AND SETTING: A retrospective review of thyroid operations performed at a tertiary referral hospital between January 2004 and December 2008. METHODS: Pathology reports were reviewed to identify the specimens that included parathyroid tissue and underlying thyroid pathology. Postoperative calcium levels were reviewed in these patients. RESULTS: During the study period, 287 thyroidectomies were performed and 47 (16.4%) patients had incidentally removed parathyroid glands. Risk factors for inadvertent parathyroid resection included total thyroidectomy (P=.0001), Hashimoto thyroiditis (P=.004), and extrathyroidal spread (P=.0003). Postoperative hypocalcemia occurred in 18 (38.3%) of the patients in whom the parathyroid gland was removed inadvertently and in 48 (20%) of the rest of the patients (P=.0123). CONCLUSION: The incidence of incidental removal of parathyroid tissue during thyroidectomy is 16.4%. Total thyroidectomy, extrathyroidal extension of the tumor, and thyroiditis were found to be the risk factors. Hypocalcemia was significantly higher among patients who had inadvertent parathyroidectomy. PMID- 21623058 TI - Survival of living-related kidney graft recipients in the era of modern immunosuppressive treatment. AB - BACKGROUND AND OBJECTIVES: Currently, there is no consensus about immunosuppressive therapy following kidney transplantation. Acute rejection rates and allograft survival rates are the clinical outcomes traditionally used to compare the efficacy of various immunosuppressive regimens. Therefore, we conducted this study to evaluate whether patient survival rates improved in the era of modern immunosuppressive treatment during living-related kidney transplantation. DESIGN AND SETTING: Retrospective cohort study in a university based tertiary internal medicine teaching hospital performed between 1999 and 2009 and patients followed up to 7 years. PATIENTS AND METHODS: Survival rates were assessed in 38 patients receiving basiliximab and mycophenolate mofetil (regimen A) and 32 patients receiving antithymocyte globulin and azathioprine (regimen B). The rest of the regimen (cyclosporine A and steroids) remained the same. A secondary end point was acute rejection episode. RESULTS: Seven-year survival rates were 100% and 72% (P=.001) and 7-year acute rejection-free survival rates were 82% and 53% (P=.03), in groups A and B, respectively. CONCLUSION: Long-term survival after living-related kidney transplantation has improved in the era of modern immunosuppressive treatment. PMID- 21623059 TI - Ambiguous genitalia: two decades of experience. AB - BACKGROUND AND OBJECTIVES: Ambiguous genitalia is a complex, medical and social emergency. The aim of this study is to present our experience over two decades, focusing on the pattern and clinical presentation. DESIGN AND SETTING: A retrospective study conducted in the pediatric endocrine clinic at a university hospital Saudi Arabia during the period 1989-2008. PATIENTS AND METHODS: Medical records of children with ambiguous genitalia were reviewed and the genitalia described. RESULTS: Of the 81 children with ambiguous genitalia, 53 (65.4%) patients were genetically females (46XX), with congenital adrenal hyperplasia being the common cause in 51 (96.5%) patients. Hyperpigmentation, variable degrees of salt wasting and a family history of a similar problem helped in diagnosis. Male genetic sex (46XY) was present in only 28 (34.6%) patients with a diversity of causes; multiple congenital anomalies in 9 (32.1%), local anorectal anomalies in 2 (7.1%), congenital adrenal hyperplasia (3-beta-hydroxysteroid dehydrogenase deficiency) in 2 (7.14%), 5-alpha-reductase deficiency in 4 (14.28%), partial androgen insensitivity in 3 (10.7%), complete androgen insensitivity in 4 (14.28%), and hypogonadotrophin deficiency in 4 (14.3%).Twenty five (47.2%) of females were wrongly assigned as males, where only two (7.1%) males were wrongly assigned as females. CONCLUSION: Ambiguous genitalia, currently termed disorders of sex development (DSD), is not uncommon in our community. Increased awareness, a detailed history, and a careful physical examination, coupled with appropriate laboratory and radiological investigations aid in early diagnosis and avoid serious sequelae. PMID- 21623060 TI - Epidemiology of sickle cell disease in Saudi Arabia. AB - Sickle cell disease (SCD) is an autosomal recessive disorder characterized by production of abnormal hemoglobin S and is associated with high morbidity and mortality. Information about the prevalence of SCD in Saudi Arabia is patchy and probably underestimated, but studies have reported that SCD is a relatively common genetic disorder in this part of the world. The prevalence of SCD in Saudi Arabia varies significantly in different parts of the country, with the highest prevalence is in the Eastern province, followed by the southwestern provinces. The reported prevalence for sickle-cell trait ranges from 2% to 27%, and up to 2.6% will have SCD in some areas. Clinical and hematological variability exists in SCD in Saudi Arabia with two major phenotypes: a mild phenotype and a severe phenotype. Further studies on the prevalence, molecular and clinical epidemiology of SCD may help predict disease severity and risk stratification of patients to determine whether to receive early intensive care or continued symptomatic care. PMID- 21623061 TI - Percutaneous vertebroplasty complications. AB - Percutaneous vertebroplasty, among various other options, has become a mainstay in the management of osteoporotic and malignant vertebral fractures. The purpose of this article is to describe complications arising from the procedure, which can be classified as mild, which may include a temporary increase in pain and transient hypotension; moderate, including infection and extravasation of cement into the foraminal, epidural or dural space; and severe such as cement leakage in the paravertebral veins, leading to pulmonary embolism, cardiac perforation, cerebral embolism or even death. Vertebroplasty is not a procedure without complications. The article defines them and describes methods to minimize them. PMID- 21623062 TI - Recurrent abdominal pain in a 16-year-old girl. PMID- 21623063 TI - Comment on: Early cholestasis in neonatal lupus erythematosus. Ann Saudi Med 2011; 31: 80-2. PMID- 21623064 TI - Comment on: Prevalence of diabetes mellitus in a Saudi community. Ann Saudi Med 2011;31(1):19-23. PMID- 21623065 TI - Comment on: Outcome of a newborn hearing screening program in a tertiary hospital in Malaysia: the first five years. Ann Saudi Med 2011; 31: 24-8. PMID- 21623066 TI - Improper aortic sinus-connected coronary artery: ascertaining the course is essential. Comment on Ann Saudi Med 2010;1:81-3. PMID- 21623067 TI - Atrial fibrillation in Saudi patients. PMID- 21623068 TI - Uterine mesenchymal tumors. AB - Uterine mesenchymal tumors are a heterogeneous group of neoplasms that can frequently be diagnostically challenging. Differentiation between the benign and malignant counterparts of mesenchymal tumors is significant due to differences in clinical outcome, and the role of the surgical pathologist in making this distinction (especially in the difficult cases) cannot be underestimated. Although immunohistochemical stains are supportive toward establishing a final diagnosis, the morphologic features trump all the other ancillary techniques for this group of neoplasms. This review therefore emphasizes the key morphologic features required to diagnose and distinguish uterine mesenchymal tumors from their mimics, with a brief description of the relevant immunohistochemical features. PMID- 21623069 TI - Conjunctivitis in the newborn- a comparative study. AB - BACKGROUND: Conjunctivitis of the newborn is defined as hyperemia and eye discharge in the neonates and is a common infection occurring in the neonates in the first month of life. In the United States, the incidence of neonatal conjunctivitis ranges from 1-2%, in India, the prevalence is 0.5-33% and varies in the world from 0.9-21% depending on the socioeconomic status. AIM: To study the organisms causing conjunctivitis of the newborn and to correlate the etiology with the mode of delivery. DESIGN: Single center, prospective, observational study. MATERIALS AND METHODS: A total of 300 mothers and their newborns, born over a period of one year, were included in the study. Of these 200 newborns were delivered through vaginal route (Group A) and 100 (Group B) delivered by lower segment caesarean section (LSCS). At the time of labour, high vaginal swabs were taken from the mothers. Two conjunctival swabs each from both eyes of the newborn were collected at birth and transported to Microbiology department in a candle jar immediately. RESULTS: Eight babies in Group A, developed conjunctivitis at birth. None of the babies in Group B developed conjunctivitis, this difference was statistically highly significant (P<0.000). The organisms found in the conjunctiva of the newborns in Group A were Coagulase negative Staphylococcus, alpha hemolytic Streptococcus, Escherichia coli and Pseudomonas spps. However, the commonest organism leading to conjunctivitis in the newborn in this study was Coagulase negative Staphylococcus. It was observed that the mothers of 5 out of 8 babies (60%) developing conjunctivitis gave history of midwife interference and premature rupture of membranes so the presence of risk factors contribute to the occurrence of conjunctivitis in the newborn. CONCLUSIONS: It is inferred that the mode of delivery and the presence of risk factors is responsible for conjunctivitis in the newborn. PMID- 21623070 TI - A decade long experience of anti-neutrophil cytoplasmic antibody testing in a tertiary care referral center in North India: perspective from a developing country. AB - BACKGROUND: In a developing, tropical country like India, discontinuous power supply, high temperatures during summer, and lack of consistent cold chain and funds provide a challenging atmosphere for anti-neutrophil cytoplasmic antibody (ANCA) testing and reporting. However, a simple in-house test and testing algorithm are described here, which have been developed and tested over time. MATERIALS AND METHODS: An analysis of a decade of testing and reporting of ANCA in the Department of Immunopathology in a tertiary referral health care center was performed to highlight the importance of testing for ANCA in proposed 1999 guideline recommended indications. RESULTS: A total of 4,195 ANCA tests were conducted from 2000 to 2009. Overall, 2060 (49%) requests had indications which met the 1999 guidelines, while the remaining 2135 (51%) fell outside the guidelines. A total of 350 samples (8.3%) were positive for ANCA on indirect immunofluorescence (IIF), out of which 212 were guideline recommended and 138 (3.2%) were non-guideline recommended ANCA requests; thus, 3.2% of non-small vessel ANCA associated vasculitis (non-SVAAV) conditions showed false positive results when the population was otherwise unselected. Maximum requests (1432) were for rapidly progressive renal failure/acute renal failure. CONCLUSIONS: The audit shows that compliance with guidelines for ANCA testing would decrease the number of false positive results. In-house screening for ANCA by IIF is cost effective and must be performed at least twice on two different samples from the same patient or on two different sets of ANCA preparations in all the cases who requested ANCA testing with a proposed 1999 guideline recommended indication. PMID- 21623071 TI - Immunohistochemical phospho tensin tumor suppressor gene staining patterns in endometrial hyperplasias: a 2-year study. AB - BACKGROUND: Endometrial carcinoma is a common neoplasm associated with the female genital tract with considerable morbidity. Endometrial hyperplasias have been widely regarded as precursor lesions. It is of importance to the pathologist to identify the subset of hyperplasias or the associated factor which could be a possible forerunner of malignancy. Phospho tensin gene (PTEN) has gained importance as one of the factors responsible. AIM: To determine the variability in PTEN expression patterns in different types of endometrial hyperplasias. SETTINGS AND DESIGN: The study was undertaken on samples received at the Department of Pathology from 2005 to 2007. MATERIALS AND METHODS: One hundred samples with 76 showing hyperplasias of different types formed the core "study group" with simple hyperplasia without atypia predominating. The rest belonged to the control groups. PTEN intensity and percentage positivity, variability in patterns of glandular and stromal expression, the number and type of PTEN null glands in different types of hyperplasia were evaluated. Statistical analyses used were Fisher's exact test based on Monte Carlo test and chi-square test. RESULTS: Complex hyperplasia was associated with a reduction in number of strongly PTEN positive glands, with an increase in null glands, seen in clusters. Co-existing atypia was associated with the weakest staining and in fewer glands. CONCLUSIONS: PTEN expression in endometrial hyperplasias can be used as an early warning of heightened cancer risk and a potential target for preventive treatment. However, extensive research is needed along this line to conclusively establish its effectiveness. PMID- 21623072 TI - Migration and maturation pattern of fetal enteric ganglia: a study of 16 cases. AB - AIMS: To study the migration and developmental pattern of ganglion cells in fetuses aged 9-21 weeks, and to document whether the migration was occurring circumferentially equally in the entire axis or if there were discrepancies in different portions at the same level. SETTINGS AND DESIGN: The hypothesis regarding the pathogenesis of Hirschsprung's disease mainly revolves around two schools. One is the single gradient migration of ganglia and the other is a dual gradient migration theory. Understanding the embryological development of enteric ganglia is necessary to study the pathogenesis of intestinal innervation disorders. MATERIALS AND METHODS: We studied the development of intestinal ganglia in fetuses aged 9-21 weeks. Serial longitudinal sections from the colon were studied, the first one including the squamo-columnar junction, for the presence and the nature of ganglion cells with Hematoxylin and Eosin, and neurone specific enolase immunostaining. Transverse sections from proximal gut were studied in a similar fashion. Thus, we evaluated the migration pattern as well as the nature of ganglia in the fetuses. We also measured the length of distal aganglionic segment in these growing fetuses. RESULTS: We noted that ganglion cells appear first in the myenteric plexus followed by deep and superficial submucous plexus. We also found evidences in favor of dual migration theory, and the distal aganglionic segment varies around the circumference of the rectal wall. CONCLUSIONS: We got evidences in support of a dual migration pattern of intestinal ganglion cells. The level of distal aganglionic segments when measured from squamo-columnar junction varied with the age of gestation and the length was incongruous. The description of distal aganglionic segment may help surgeons while taking biopsies or during operative procedures. PMID- 21623073 TI - Expression of erythropoietin and its receptor increases in colonic neoplastic progression: the role of hypoxia in tumorigenesis. AB - BACKGROUND: Tissue hypoxia is a characteristic patho-physiologic property of colorectal cancer. This process may also add to a therapeutic problem of solid tumor resistance to chemo- and radiation therapy. Erythropoietin (Epo) expression is induced by tissue hypoxia. Acting via its receptor (EpoR), Epo inhibits apoptosis of erythroid cells and has been shown to rescue neurons from hypoxic damage. Increased Epo and EpoR expression has been recently described in human breast, renal and cervical carcinoma. Given the characteristic tumor diathesis present in majority of colorectal cancers, we examined whether Epo signaling may play a role in colonic neoplastic progression. MATERIALS AND METHODS: Expression of Epo and EpoR was examined using immunohistochemistry in 24 cases of primary colorectal and metastatic adenocarcinomas versus adenomas and normal colonic mucosa. Immunohistochemical stains were evaluated semiquantitatively based on a four-tiered scale. Based on the combination of extent and intensity of immunoreactivity, an immunostaining score (0-300) was determined for each sample. Expression of Epo and EpoR protein and mRNA was examined using Western blot and reverse transcriptase-polymerase chain reaction (RT-PCR), respectively, in both normal colonic tissue and carcinoma specimens in five cases. RESULTS: Epo expression was sequentially increased in normal colonic mucosa (8.3 +/- 5.6, mean +/- SEM), adenoma (26.4 +/- 9.1), primary carcinoma (96.1 +/- 12.8) and metastatic carcinoma (122 +/- 51.3). EpoR expression was also sequentially increased in normal colonic mucosa (22.3 +/- 11.8), adenoma (108.7 +/- 24.2), primary carcinoma (178.7 +/- 16.6) and metastatic carcinoma (220 +/- 58.3) (P< 0.05 for all results). Epo and EpoR showed enhanced expression in the areas adjacent to ischemia/necrosis. Western blot and RT-PCR analysis revealed increased EpoR protein and mRNA levels in carcinoma compared to normal mucosal colon specimens. Focal stromal Epo and EpoR immunoreactivity was present in 10 and 12 cases, respectively. CONCLUSIONS: The uniform increase in the expression of Epo and EpoR along the colonic neoplastic sequence and further increase in ischemic/necrotic areas indicates that the Epo signaling pathway is an important component in colon carcinogenesis including possible epithelial-stromal interactions. PMID- 21623074 TI - Comparative study of histopathological Marsh grading with clinical and serological parameters in celiac iceberg of north India. AB - BACKGROUND: Celiac disease is an autoimmune disorder caused by the ingestion of wheat gluten and related proteins in genetically susceptible individuals. It is characterized by anti-tissue transglutaminase (anti-tTG) antibodies. Duodenal biopsy is the gold standard for diagnosis. Correlation of clinical, serologic, and histological features is essential for a definitive diagnosis. The ratio of diagnosed versus undiagnosed cases is quite high. AIMS: This study aimed to correlate the degree of mucosal damage with anti-tTG levels, mean baseline hemoglobin and endoscopic findings. SETTING AND DESIGN: Two hundred twenty six adults suspected to have celiac disease were studied. Marsh grades were compared with anti-tTG levels, hemoglobin, endoscopy, and clinical presentations. MATERIALS AND METHODS: Esophagogastroduodenoscopy, serum levels of anti-tTG, complete hematologic work-up, and duodenal biopsy were performed in all 226 cases (including three siblings of confirmed patients) with well-defined symptom groups. Histopathological grading was done as per modified Marsh system. Correlation of all the parameters was performed with Marsh grades. STATISTICAL ANALYSIS: Performed on SPSS version 15.0. Tests applied include one way ANOVA, Chi-square test, repeated measure analysis, and Bonferroni's method for comparison. Results were considered significant when P<0.05. RESULTS AND CONCLUSIONS: Anti-tTG levels, mean baseline hemoglobin, and endoscopic findings were found to correlate with increasing severity of mucosal damage with P<0.001 for all. Anti-tTG levels of grades 1+2 and those of grade 3a were significantly different from levels of grades 3b and 3c+4 with P<0.001 for each. Varied clinical presentations of celiac disease were seen in the adult wheat eaters of North India. PMID- 21623075 TI - Expression of human mammaglobin and clinicopathologic correlations in breast cancer: the findings in Malaysia. AB - BACKGROUND: Human mammaglobin (hMAG) is a secreted protein which has been detected in breast epithelial cells of mammary glands and has been used as a specific marker for breast cancer. OBJECTIVES: This study aims at studying the hMAG expression and identifying the significant predictors of hMAG expression in breast cancer tissues. MATERIALS AND METHODS: The tissue samples were obtained from two major teaching hospitals in the country. They were examined by immunohistochemistry (IHC) and the hMAG expression was evaluated using an established scoring system. RESULTS: Out of 84 breast cancer tissue samples, hMAG was expressed in 50 samples (59.6%). The expression of hMAG was found to be increased with cancer grade. The output of logistic regression model showed that hMAG was overexpressed in breast cancer samples from the first hospital (P = 0.014), but not with those from the second hospital. CONCLUSIONS: It can be concluded that hMAG may serve in the diagnosis and the assessment of progression with the increased cancer grade. The dominance in hMAG expression in samples from HUSM may correlate with ethnic, environmental or genetic factors. PMID- 21623076 TI - Strong BCL2 expression in Burkitt lymphoma is not uncommon in adults. AB - BACKGROUND: The distinction between Burkitt lymphoma (BL) and diffuse large B cell lymphoma (DLBCL) is not always easy, so much so that the WHO 2008 Blue Book has incorporated a provisional category of "B-cell lymphoma, unclassifiable with features intermediate between DLBCL and BL." One of the immunohistochemical (IHC) markers used at times to differentiate between the two is IHC expression of BCL2, which was initially believed to be consistently negative in BL. Later it was recognized that weak BCL2 expression is still compatible with the diagnosis of BL. To verify or otherwise this current view this study was undertaken. MATERIALS AND METHODS: We retrieved 39 confirmed cases of BL, in both children and adults. All these cases had typical morphology, IHC profile, and Mib-1 index that are typical of BL. All these cases were then stained with a monoclonal antibody against BCL2 oncoprotein, using 2-step Envision system. RESULTS: Out of 39 cases, 31 cases (79.4%) were completely negative for BCL2, whereas 5 (12.8%) were weak focal positive. However, another 4 (10.2%) cases did show strong diffuse cytoplasmic staining for BCL2. Fluorescent in-Situ hybridization (FISH) for t(14:18) was optimally done on 6 out of 9 cases. All these 4 cases were from adults with 3 out of 4 arising in the parotid region. Two out of 4 cases also showed t(8:14) on FISH. CONCLUSIONS: It was concluded that contrary to the common belief, strong BCL2 IHC expression is possible in typical BL in adults and cannot be absolutely relied upon to distinguish between BL and DLBCL. PMID- 21623077 TI - Immunophenotypic profile of plasma cell leukemia: a retrospective study in a reference cancer center in India and review of literature. AB - BACKGROUND: Plasma cell leukemia (PCL) is a rare but aggressive subtype of plasma cell dyscrasia. It is known to present with highly variable morphological features and may mimic with other lymphoid neoplasms. Multicolor flow cytometry (MFC) with availability of newer markers is highly useful in the diagnosis of the plasma cell leukemia. We present an immunophenotypic profile in ten cases of PCL along with their clinical and laboratory findings. MATERIALS AND METHODS: We retrospectively studied immunophenotypic profile of 10 cases of plasma cell leukemia (out of 4615 cases of hematolymphoid neoplasms) using five parameter, three color flow cytometric analysis. We also studied their clinical presentation and other laboratory findings. RESULTS: Common clinical features at presentation were weakness, bone pain, anemia, thrombocytopenia and osteolytic lesions. Plasma cell population was identified on strong expression of CD38 and co-expression of CD38 and CD138. CD56 was expressed in 44% cases. CD19 and CD20 were negative in all cases. Surface light chain restriction was seen in 50% cases and in remaining 50% cases revealed cytoplasmic light chain restriction. CD117 was expressed in one out of two cases studied. CONCLUSIONS: MFC immunophenotyping is highly useful to differentiate Plasma cell leukemia from other chronic lymphoproliferative disorders with plasmacytoid morphology as well as from non-neoplastic reactive PC and co-expression of CD38 and CD138 is a best combination to identify the plasma cells by MFC. PMID- 21623078 TI - Frequency of central nervous system tumors in delta region, Egypt. AB - INTRODUCTION AND AIM OF WORK: Central nervous system (CNS) tumors represent a major public health problem, and their epidemiological data in Egypt have been rather incomplete except for some regional reports. There are no available frequency-based data on CNS tumors in our locality. The objective of this study was to estimate the frequency of CNS tumors in east delta region, Egypt. MATERIALS AND METHODS: The data were collected during the 8-year period from January 1999 to December 2007 from Pathology Department, Mansoura University, and other referred pathology labs. Examination of HandE stained sections from retrieved paraffin blocks were done in all cases for histopathologic categorization of C.N.S. tumors. Immunohistochemical studies were applied to confirm final histopathologic diagnosis in problematic cases. RESULTS: Intracranial tumors represented 86.7% of cases in comparison to only 13.3% for spinal tumors. Gliomas were the CNS tumors of the highest frequency (35.2%), followed by meningioma (25.6%), pituitary adenoma (11.6%) and nerve sheath tumors (6.6%). 10.25% of tumors were of children <15 years. CONCLUSION: This study provides the largest series of the relative frequency of CNS tumors in Delta region in Egypt till now and may help to give insight into the epidemiology of CNS tumors in our locality. PMID- 21623079 TI - Ki-67 and Bcl-2 in colorectal polyps with breast cancer. AB - BACKGROUND: The most common second primary cancer in women who survive breast cancer is colorectal cancer. Population cohort studies suggest that the risk of colorectal cancer is also high in female relatives of women with breast cancer compared to the general population. The histological distinction between a large hyperplastic polyp and a serrated adenoma is not straightforward and there is probably no sharp division between hyperplastic polyposis and 'serrated adenomatous polyposis'. AIM: The aim of the study was to define the distribution of Ki-67 and Bcl-2 protein in hyperplastic colorectal polyps of women with breast cancer. MATERIALS AND METHODS: A cross-sectional, controlled study was conducted in 40 women with and without breast cancer who had hyperplastic colorectal polyps. The patients were divided into two groups: Group A (a control group of women without breast cancer, n = 20) and Group B (a study group of women with breast cancer, n = 20). The expression of Ki-67 and Bcl-2 protein was assessed on the basis of quantity of immunostaining, by counting antigen positive cells, in comparison with normal mucosa. Student's t-test and the chi-square test were used to analyse Ki-67 and Bcl-2 expression, respectively. RESULTS: Statistical significance was established at P < 0.05. The mean percentage of Ki-67 stained nuclei in Groups A and B was 25.11 +/- 2.07 and 41.51 +/- 1.86, respectively (P < 0.003), whereas the percentage of cases with cells expressing Bcl-2 in Groups A and B was 15% and 85%, respectively (P < 0.002). CONCLUSIONS: In the present study, greater proliferative activity and greater expression of the antiapoptotic protein Bcl-2 was found in the hyperplastic colorectal polyps of women with breast cancer. PMID- 21623080 TI - Duodenal nodularity in children: a clinical and pathologic study of 17 cases. AB - AIMS: Duodenal nodularity is an uncommon endoscopic appearance of numerous visible mucosal nodules in the proximal duodenum. In this retrospective study we aimed to determine the clinical significance and histopathologic features of duodenal nodularity in children. MATERIALS AND METHODS: The medical records of the patients who were defined to have duodenal nodularity by endoscopy were reviewed. STATISTICAL ANALYSIS USED: The data were expressed as mean +/- SD and percentages (%). RESULTS: Seventeen patients with endoscopically defined duodenal nodularity were chosen. The mean age at diagnosis was 12.1 years (range: 6-17 years), 9 males. Abdominal pain (47%) was the most common clinical symptom and antral nodularity (41%) was the most common endoscopic finding in children with duodenal nodularity. Histopathologic evaluation of duodenal nodules revealed chronic inflammation in all patients, increased intercryptal and intraepithelial numbers of eosinophils in 70.5%, and villous atrophy in 47% of patients. Giardia infestation was demonstrated in 6 patients by histologic examination and/or Giardia lamblia-specific antigen positivity in stools. The clinical diagnoses of the patients have shown variations, such as celiac disease, giardiasis, secretory IgA deficiency, and Helicobacter pylori gastritis, and some of them were associated with the others. CONCLUSIONS: Although the endoscopic appearance is similar, clinical spectrum and pathologic features are not so similar and there are no specific histomorphologic findings for nodularity. The most demonstrative findings we observed in children were increased lymphocyte and/or eosinophil infiltration in the duodenal mucosa. We suggested that care should be taken in the evaluation of microbiological and immunologic etiologies causing this prominent inflammatory reaction. PMID- 21623081 TI - Subcutaneous panniculitis-like T-cell lymphoma: a clinicopathologic study of 5 cases. AB - BACKGROUND: Subcutaneous panniculitis-like T-cell lymphoma is as yet a poorly characterized subtype of cutaneous T-cell lymphomas. It is difficult to diagnose and lacks a standard treatment protocol. MATERIALS AND METHODS: We report our experience with five such cases in patients with age ranging from 13 to 45 years, wherein we reviewed their clinical presentations, histopathological findings, treatment and clinical outcome. RESULTS: Three out of the five cases are alive of which two are in complete remission. Our series stresses on the need to be aware of this uncommon entity in a dermatology clinic. Facial and upper extremity skin nodules with involvement of the lower dermis on light microscopical examination and suggestive immunohistochemical findings were frequently observed in our patients in contrast to previously described cases. PMID- 21623082 TI - Profile of H1N1 infection in a tertiary care center. AB - BACKGROUND: A novel swine origin influenza virus (H1N1) is spreading worldwide and threatens to become pandemic.H1N1 critical illness mostly affects young patients and is often fatal. AIM: The aim of the present study is to evaluate the clinical characteristic of H1N1 infection in a tertiary care hospital. MATERIALS AND METHODS: A total of 92 nasal and pharyngeal swabs from suspected cases of swine flu were processed by real time reverse transcriptase polymerase chain reaction (rRT-PCR). RESULT: Twenty(21.73%) were positive of which two were treating physicians and five (25%) patients expired. CONCLUSIONS: The age group of positive cases of H1N1 was between 21 and 30 years and age group of patients who died ranged from 40 to 45 year. This overview indicates that although the majority of hospitalized persons infected with novel influenza A (H1N1) recovered without complications, certain patients had severe and prolonged disease. It was also noted that 2009 influenza A (H1N1) infection - related clinical illness predominantly affects young patients. All hospitalized patients with novel influenza A (H1N1) infection should be monitored carefully and treated with antiviral therapy. Mandatory vaccination of health-care workers is especially important in emerging pandemic. PMID- 21623083 TI - Role of PCR for diagnosing Pneumocystis jirovecii pneumonia in HIV-infected individuals in a tertiary care hospital in India. AB - OBJECTIVES: In developing countries like India, the diagnosis of Pneumocystis jirovecii infection is often made either by conventional staining or clinically. This study was planned to know the utility of polymerase chain reaction (PCR) in diagnosing Pneumocystis jirovecii pneumonia (PJP) in human immunodeficiency virus (HIV)-infected patients, to compare the PCR results with that of staining techniques and also to correlate the results with clinical condition of patients. MATERIALS AND METHODS: A prospective study included 50 HIV-infected adult in patients with symptoms of lower respiratory tract infection. Induced sputum, bronchoalveolar lavage or tracheal aspirate were proceeded for both staining and PCR for mitochondrial large subunit rRNA gene of P. jirovecii. RESULTS: In our study PCR results correlated with staining findings in 14% (7/50) of cases. Another 20% (10/50) cases could be diagnosed only with PCR, where staining was negative for the presence of P. jirovecii. When compared with clinical evidence of disease, PCR showed 93.7% sensitivity and 94.1% specificity. Presence of dyspnea and CD 4 count showed statistical significance (P<0.05) in PCP-diagnosed patients. CONCLUSIONS: PCR can be used for early and accurate diagnosis of PCP in HIV-infected patients. PMID- 21623084 TI - Clonality assessment of lymphoproliferative lesions using the polymerase chain reaction: an analysis of two methods. AB - BACKGROUND: Lymphoid malignancies are a heterogeneous group of disorders which may be difficult to differentiate from reactive proliferations even after immunohistochemistry. Polymerase chain reaction (PCR) is believed to be a good adjunct tool for diagnosis. MATERIALS AND METHODS: We examined 24 cases of neoplastic and non-neoplastic lymphoproliferative lesions in this study and evaluated the PCR as an additional tool in the confirmation of the diagnosis. Two different PCR methodologies were evaluated. RESULTS: In the evaluation of the T cell PCR, it was seen that the correlation using both the commercial kits and the custom-synthesized primers was highly significant at a P value of <0.05. In the evaluation of the B-cell PCR, it was seen that the correlation using both the commercial kits and the custom-synthesized primers was not significant using either method (P > 0.05). CONCLUSIONS: Both the methods showed an excellent concordance for T-cell gamma gene rearrangements, However, the same was not seen in the B-cell receptor rearrangements. This may be because of the small sample size or the inability of consensus V primers to recognize complementary DNA sequences in all of the V segments. PMID- 21623085 TI - Immunohistochemical and morphologic findings in columnar cell lesions of the breast. AB - BACKGROUND: Columnar cell lesions (CCLs) with or without atypia frequently coexist with invasive or in situ breast carcinomas. In this study, 39 mastectomy specimens containing CCLs coexisting with invasive carcinomas were retrospectively analyzed for cellular characteristics and structural pattern of CCL neighboring the tumor. MATERIALS AND METHODS: The expression of estrogen receptor (ER), progesterone receptor (PR), and p53 antibodies in CCL and coexisting invasive tumors, type of invasive tumor, histopathologic grade, and presence of atypia in CCL have been studied. RESULTS: Sixteen (41%) of all CCLs were with atypia, whereas 23 (59%) of them were without atypia. No correlations were found between the presence of CCLs with atypia and either the morphologic type of carcinoma or histopathologic grade of the tumors. Presence of atypia in the CCL was not correlated with the expression of p53 in the invasive tumors. CCLs without atypia dominated in Grade III tumors. The percentages of CCLs without atypia were also higher in both ER (-) and PR (-) tumors. CONCLUSIONS: CCL with atypia is generally considered to be a precursor of invasive carcinoma; however, in our study, CCLs without atypia more frequently coexisted with breast carcinoma. PMID- 21623086 TI - Viral ventilator-associated pneumonia: uncovering tip of the iceberg. AB - CONTEXT: Hospital-acquired infections are frequently encountered by the physicians for ailments demanding prolonged hospitalization, especially in intensive care units, where patients are often mechanically ventilated. The organisms most often implicated are bacteria; viral etiology is infrequent. AIMS: The study aims at reviewing lung pathology at autopsy in mechanically ventilated children admitted in pediatric intensive care unit (PICU) to assess the incidence of viral ventilator-associated pneumonias (VAP). SETTING AND DESIGN: Retrospective analysis. MATERIALS AND METHODS: Among the 275 children who had been autopsied, 13 who had been admitted in the PICU satisfied the criteria for VAP. These cases were analyzed on the basis of clinical data and pulmonary pathology. Depending on the overall histology, the cases were classified as being viral or bacterial in etiology. Immunohistochemistry (IHC) for detection of viral antigens was also performed. RESULTS: Of the 13 children, nine (five males and four females) had shown the histomorphologic features, suggesting viral inflammation. The mean age was 33 months. Falling oxygen saturation and increasing respiratory distress had necessitated ventilator support. Acute lymphocytic bronchiolitis, interstitial pneumonitis, diffuse alveolar damage, and necrotizing pneumonia were the histological features. The viruses identified in five patients were adenovirus, respiratory syncytial virus and cytomegalovirus. CONCLUSION: This communication, though not representing the true incidence, emphasizes that a proportion of nosocomial infections is due to viral infections. This should alert the treating intensivists to actively pursue investigations to confirm viral etiology. PMID- 21623087 TI - Fungal infections as a contributing cause of death: an autopsy study. AB - CONTEXT: With the continuing rise in the number of immunocompromised patients, the incidence of invasive mycoses has increased. Various studies have reported the trends of fungal infections in autopsies. Because of limitations in antemortem clinical diagnosis owing to lack of sensitive diagnostic tools, information regarding frequency and pathogenesis of fungal infections is largely dependent on autopsy studies. AIM: To study the prevalence of fungal infections at autopsy spanning a period of 20 years and to document recent trends, prevalence of various fungi over decades along with underlying predisposing factors and pathological findings. SETTINGS AND DESIGN: Retrospective study. MATERIALS AND METHODS: All autopsies between 1988 and 2007 were reviewed and all cases showing fungal infections were analyzed. The clinical details and demographic data were retrieved from medical records. Representative sections from all organs were stained with hematoxylin and eosin stain and special stains including Gomori's silver methenamine (GMS) and periodic acid Schiff (PAS). Culture details were noted, wherever available. RESULTS: A total of 401 autopsies were performed during the study period. Fungal infections were identified in 35 (8.7%) of these cases. Leukemia was the commonest risk factor. The commonest pathogen in the present study was Aspergillus sp. The commonest single organ involved was brain (n = 18). Culture positivity was seen in 23.8% cases. CONCLUSION: The study highlights various predisposing factors and organisms in autopsy series. Existing diagnostic modalities are not sensitive to ensure antemortem diagnosis of fungal infections. PMID- 21623088 TI - Dysferlinopathy: spectrum of pathological changes in skeletal muscle tissue. AB - BACKGROUND: Dysferlinopathy is an autosomal recessive-limb girdle muscular dystrophy (AR-LGMD) caused due to the defect in gene encoding dysferlin, a sarcolemmal protein. Awareness of the variants and their relative frequency is essential for accurate diagnosis. AIM: To study the spectrum of morphologic changes in immunohistochemically proven cases of dysferlinopathies, to correlate the findings with clinical phenotype and durations of illness and determine the frequency. MATERIALS AND METHODS: Dysferlinopathies seen over a period of 2 years at a tertiary neurological center were analyzed. RESULTS: Clinically, majority had Miyoshi phenotype (46.6%) with distal involvement and LGMD phenotype (40%) with proximal muscle involvement. In addition, a proximo-distal and tibial muscle phenotype was encountered. Morphologically, rimmed vacuoles were noted in the Miyoshi phenotype. The presence of ragged red fibers, lobulated fibers and inflammation had no preference to a particular phenotype. Significant atrophy and lobulated fibers were noted in patients with longer duration of illness. CONCLUSIONS: Dysferlinopathy was the second most common identifiable cause (21%) of LGMD next to sarcoglycanopathies (27%). PMID- 21623089 TI - Relation of hepatitis B core antigen expression with histological activity, serum HBeAg, and HBV DNA levels. AB - BACKGROUND: Hepatitis B virus (HBV) is a potentially life-threatening liver infection which may progress to liver failure and cirrhosis. Intrahepatic expression patterns of viral antigens detected by immunohistochemistry may have prognostic implications in disease process. AIM: In this study, we aimed to investigate the relationship between the HBV core antigen (HBcAg) expression and histological activity index (HAI), fibrosis, serum hepatitis B e-antigen (HBeAg) status and HBV DNA levels in patients with chronic HBV infection. MATERIALS AND METHODS: A total of 114 liver biopsies from patients with chronic HBV infection were included in the study. Immunohistochemical expression of HBcAg and its relation with HAI, fibrosis, serum alanine aminotransferase (ALT) levels, HBeAg status and HBV DNA levels were assessed. RESULTS: The presence of nuclear expression of HBcAg did not show any correlations with ALT levels, HAI and fibrosis score. When the groups were categorized according to the HBeAg status, nuclear HBcAg expression was found to be high in HBeAg positive patients. However, HBcAg nuclear expression showed significant correlations with HBV DNA levels and fibrosis scores in HBeAg negative but not HBeAg positive patients. HBV DNA levels were also significantly associated with HAI and fibrosis scores in HBeAg negative patients. CONCLUSIONS: Significant differences found between HBeAg positive and negative patients suggest that HBeAg negative disease is different from HBeAg positive disease, and also point outs that in HBeAg negative disease, patients with nuclear HBcAg expression and increased levels of HBV DNA levels are at a higher risk of developing progressive liver disease. PMID- 21623090 TI - Detection of HIV drug resistance mutations in pregnant women receiving single dose Nevirapine in south India. AB - BACKGROUND: Single dose of Nevirapine to prevent mother to child transmission of HIV is the commonest preventive regimen in resource-limited countries. OBJECTIVES: The objective of this study was to detect drug-resistant virus after single dose of Nevirapine (sdNVP) provided to delivering HIV seropositive (HIV+ve) women and to evaluate the time taken for its decay. RESULTS: Of the 36 consenting HIV+ve pregnant women enrolled into the study, the mean hemoglobin and total lymphocyte counts were 10.8 g/dl and 1843 cells/mm 3 , respectively. Mean CD4 counts in 64% of women was 363 cells/mm 3 and mean viral load for 16/36 women was 28,143 copies/ml of plasma. Nevirapine-resistance mutations were detected in 28% of women at delivery; using OLA (Oligonucleotide Ligation Assay). K103N mutations were seen in 19.4% of women while the Y181C mutation was seen in 5%. Both the mutations were detected in 2.7% of women. Sequential blood samples collected at delivery, 7-10 days, 6 weeks, 4 months, 6 months and one year postpartum showed that 81% of K103N mutations and 66.7% of Y181C mutations were detected at 6 weeks postpartum . Wild-type virus had replaced the mutants by one year postpartum in all women except one. CONCLUSION: These observations are relevant for future treatment with antiretroviral therapy in these women for their HIV disease. PMID- 21623091 TI - Gastrointestinal stromal tumor with skeletal muscle, adrenal and cardiac metastases: an unusual occurrence. AB - Gastrointestinal stromal tumor (GIST) is the most common mesenchymal tumor of the bowel. It most commonly arises in the stomach, followed by the small intestine, colorectum and the esophagus. It usually spreads to the liver or the peritoneum, with bone and lung metastases being infrequent. Here, we present a case report of GIST behaving in a bizarre way. The patient presented with skeletal muscle metastasis and was initially misdiagnosed as leiomyosarcoma. On follow-up, he developed adrenal metastasis, and later, cardiac metastasis. None of the above unusual sites has previously been reported in literature to our knowledge. PMID- 21623092 TI - Adenomyoma of common bile duct arising in a type I choledochal cyst. AB - Adenomyoma can be misdiagnosed as an adenocarcinoma, leading to needless and extensive surgical resections. A 45-year-old woman presented with right hypochondrial pain. Magnetic resonance imaging showed a choledochal cyst. Excision of choledochal cyst with Roux-en-Y hepaticojejunostomy was performed. A segment of dilated common bile duct and an attached nodule was received. Sections from the choledochal cyst showed a cyst wall composed of dense fibrous tissue lined by partially ulcerated columnar epithelium. Sections from the nodule showed interlacing whorls of smooth muscle bundles with entrapped glands. The glands were lined by cuboidal to columnar cells without nuclear atypia. This was recognized as an adenomyoma. To the best of our knowledge, this is the first reported case in which an adenomyoma was found associated with a type 1 choledochal cyst. A review of the existing literature and discussion of theories of genesis and the diagnostic pitfalls are presented. PMID- 21623093 TI - Septicemia due to Capnocytophaga canimorsus following dog bite in an elderly male. AB - A previously healthy, 67-year-old, man with past medical history of myocardial infarction and hypertension was rushed to the emergency room after sudden onset of fever, chills, severe rigors, hypotension, tachycardia and vomiting. The patient was diagnosed as being in septic shock, and investigations revealed intracellular gram-negative bacilli in polymorphonuclear leukocytes in the peripheral smear. A history of dog contact was elicited after this very unusual and rare finding. Cultures confirmed septicemia due to Capnocytophaga canimorsus, a normal oral and nasal flora inhabitant of cats and dogs that can cause severe and sometimes fatal septicemia in humans. We report this very interesting case because of the common prevalence of dog homeownership and the rarity of C. canimorsus inducing sepsis. PMID- 21623094 TI - Therapy related myelodysplastic syndrome: a case report and review of literature. AB - Therapy related myeloid neoplasm is directly related to previous cytotoxic chemotherapy or radiation therapy. We present a 47-year-old lady who developed therapy related myelodysplastic syndrome (MDS) 2.5 years after she received four cycles of chemotherapy and local radiation therapy for carcinoma breast. She presented with bicytopenia with trilineage dyspoiesis in the peripheral blood, bone marrow aspirate and biopsy. Fluorescent in-situ hybridization studies did not reveal any of the common abnormalities associated with MDS. A diagnosis of therapy related MDS was rendered. Different studies have shown that patients treated with alkylating agents and ionizing radiation present as MDS with a latent period of 3-10 years. Our patient developed MDS within 2.5 years of starting chemotherapy and radiotherapy and did not reveal any of the conventional cytogenetic abnormalities. It highlights the importance of simple tests like a complete blood count and peripheral blood smear examination in follow-up of the patients treated with chemotherapy. PMID- 21623095 TI - Lafora disease: a case report, pathologic and genetic study. AB - A 19-year-old male patient presented with progressive myoclonic seizures and speech disorder. The patient had photosensitivity, a few episodes of sudden transient blindness, and infrequent complex visual auras, dysarthria and mild ataxia, frequent myoclonic jerks prominently in the legs and severe dementia. Microscopic examination of the axillary skin biopsy revealed periodic acid-Schiff positive inclusion bodies in abluminal side of the apocrine sweat gland acini. Molecular screening showed a homozygous R241X mutation in EPM2A. Genotyping helps in the correct diagnosis of the Lafora disease (LD), which may be difficult to diagnose based on the available histopathological testing only. Our study is an effort to determine the distribution of mutations in LD patients in our region. PMID- 21623096 TI - Subcutaneous panniculitic-like T-cell lymphoma: a red alert! The role of a vigilant histopathologist. AB - Subcutaneous panniculitic-like T-cell lymphoma constitutes a distinctive clinicopathologic entity derived from cytotoxic T lymphocytes. A 25-year-old female presented with fever and skin lesions over the upper limb, lower limb and trunk since 2 years. On examination, there were multiple subcutaneous, tender, erythematous, poorly circumscribed indurated plaques and nodules on the upper limbs and lower limbs. Histopathological examination revealed subcutaneous fat displaying a predominantly lobular infiltration of atypical lymphoid cells. Characteristically, there was rimming of individual fat cells by the surrounding neoplastic lymphocytes. Immunohistochemical evaluation of the neoplastic lymphocytes showed CD3 and CD5 immunoreactivity and CD30 and CD20 negativity. A diagnosis of subcutaneous panniculitic T-cell lymphoma was made. SPTCL is a rare cytotoxic lymphoma that can be misdiagnosed as benign panniculitis due to similarities in clinical and histological features between the two entities and thus cause a diagnostic hindrance. PMID- 21623097 TI - Refractory anemia leading to renal hemosiderosis and renal failure. AB - Renal hemosiderosis is a rare cause of renal failure and, as a result, may not be diagnosed unless a detailed history, careful interpretation of blood parameters and renal biopsy with special staining is done. Here, we present a rare case of renal hemosiderosis presenting with renal failure. PMID- 21623098 TI - High ferritin and myoglobin level in legionella pneumonia: a case report and review of literature. AB - Legionella pneumophila is a cause of both community- and hospital-acquired pneumonia and might cause high morbidity and mortality. Therefore, early diagnosis and treatment with appropriate antibiotics is crucial. Many clinical and laboratory abnormalities can be observed in the course of Legionella pneumonia. In this study, we aimed to present simultaneously increased serum ferritin and myoglobin level in a legionella case with reference to the relevant literature. PMID- 21623099 TI - Poorly differentiated monophasic synovial sarcoma of the mediastinum. AB - Poorly differentiated synovial sarcoma is a diagnostically challenging neoplasm. Most commonly they occur in the soft tissue of the extremities and are rare in the mediastinum. They can be indistinguishable from other "round cell tumors" based on the morphology alone or at times by immunohistochemical studies. Here in, we report an extremely rare case of metastatic poorly differentiated monophasic synovial sarcoma of the mediastinum without a known primary in a 30 year-old man. The imaging studies on admission showed 10 * 9.5 cm anterior mediastinal mass with multiple nodules in the lung and pleura along with multiple enlarged mediastinal and axillary lymph nodes. Histopathologic and immunohistochemical analysis supported the diagnosis of poorly differentiated synovial sarcoma, which was further confirmed by molecular genetic analysis. PMID- 21623100 TI - Oral diffuse B-cell non-Hodgkin's lymphoma associated to Gorlin-Goltz syndrome: a case report with one year follow-up. AB - Nevoid cell carcinoma syndrome or Gorlin-Goltz syndrome is an autosomal dominant disorder characterized by multiple basal cell carcinoma, multiple keratocyst tumors, and skeletal anomalies. The Gorlin-Goltz syndrome has been associated with numerous benign and malignant neoplasms. The authors describe a case of Gorlin-Goltz syndrome in association with non-Hodgkin's lymphoma. To the best of our knowledge, this is the second case described in the English literature. PMID- 21623101 TI - Mammary-type myofibroblastoma of soft tissue. AB - A 40-year-old woman presented with a 1 year history of a painless, subcutaneous lump on the right buttock. Clinical examination showed an approximately 6 cm large subcutaneous mass covered by apparently normal-looking skin. No inguinal lymphadenopathy was found. The mass was excised with the clinical diagnosis of fibroma. Histologically, the lesion was consistent with mammary-type myofibroblastoma of soft tissue, a very rare, benign mesenchymal neoplasm with myofibroblastic differentiation. After surgical excision she was free of recurrence over a period of 8 months. This article also challenges the theory that suggests the origin of this tumor to be from the embryonic mammary tissue, adding another case of a site other than the milk lines. PMID- 21623102 TI - Cutaneous metastasis of prostate carcinoma to neck and upper chest. AB - Prostate adenocarcinoma is the most common urologic malignant neoplasm in men. Metastasis to skin is rarely reported and usually occurs late. The incidence and appearance of cutaneous metastasis are not well established in patients with prostate adenocarcinoma and their recognition remains poor among practicing urologists. Their clinical appearance may mimic other common dermatologic disorders. Definitive diagnosis requires a high index of suspicion. Immunohistochemical staining helps in establishing the diagnosis. We report a case of prostate adenocarcinoma presenting with widespread metastasis, including those to dermis and subcutaneous tissue of neck and upper chest. PMID- 21623103 TI - Giant cell tumor of bone with lymph node metastases: a rare presentation. PMID- 21623104 TI - Fine-needle aspiration cytology of jugular foramen meningioma presenting as parapharyngeal mass. PMID- 21623105 TI - Sarcomatoid carcinoma of the urinary bladder: a report of two cases. PMID- 21623106 TI - Metastatic immature teratoma: a diagnostic challenge on fine-needle aspiration cytology. PMID- 21623107 TI - 'Functional' paraganglioma of ureter: an unusual case. PMID- 21623108 TI - Role of HPLC in the detection of HbH disease. PMID- 21623109 TI - Corneal abscess caused by Bipolaris spicifera. PMID- 21623110 TI - Ascertaining the prevalence of occult hepatitis B virus infection in voluntary blood donors: a study from Central India. PMID- 21623111 TI - Papillary meningioma: a rare malignant variant. PMID- 21623112 TI - Isolated hydatid cyst in the submandibular salivary gland: a rare primary presentation (diagnosis by fine needle aspiration cytology). PMID- 21623113 TI - Large ovarian leiomyoma in a postmenopausal woman. PMID- 21623114 TI - Synchronous occurrence of anaplastic, follicular and papillary carcinomas with follicular adenoma in thyroid gland. PMID- 21623115 TI - Fungal characterization using polymerase chain reaction in patients with fungal sinusitis. PMID- 21623116 TI - Infant bacteremia due to Salmonella typhimurium. PMID- 21623117 TI - When molecular diagnosis went wrong. PMID- 21623118 TI - Infection control practices need ingenuous processes beyond prescribed guidelines. PMID- 21623119 TI - A novel FRET probe-based approach for identification, quantification, and characterization of occult HCV infections in patients with cryptogenic liver cirrhosis. PMID- 21623120 TI - Early diagnosis of tuberculous meningitis: a comparison of nested polymerase chain reaction and BacT/ALERT. PMID- 21623121 TI - Primary spindle cell melanoma of the urinary bladder. PMID- 21623122 TI - Fine-needle aspiration biopsy of adenoid cystic carcinoma of the palatine tonsil. PMID- 21623123 TI - Mesenteric vessel thrombosis and hypercoagulable states. PMID- 21623124 TI - Leptospirosis-induced still birth and postpartum sepsis. PMID- 21623125 TI - Isolation of Salmonella paratyphi A from a female with diabetic foot ulcer. PMID- 21623126 TI - A case of Dirofilaria immitis presenting as an intramuscular soft tissue mass. PMID- 21623127 TI - Sjogren's syndrome complicated by non-Hodgkin's lymphoma: spectrum of lymphoid proliferations and associated hepatitis C virus infection. PMID- 21623128 TI - Bacteriology profile among patients with ventilator-associated pneumonia from a medical intensive care unit at a tertiary care center in Mumbai. PMID- 21623129 TI - Fourth-generation enzyme immunoassays for screening of HIV in blood donors: need of the hour. PMID- 21623130 TI - Comment on: synchronous occurrence of anaplastic, follicular, and papillary carcinoma with follicular adenoma in thyroid gland. PMID- 21623131 TI - Butterfly effect and cancer. PMID- 21623132 TI - Use of artisanal tissue micro-array in research involving immunohistochemistry and in situ hybridization in Hodgkin's lymphoma at University Hospital in Rio de Janeiro, Brazil. PMID- 21623133 TI - Unilateral multicystic dysplasia in an ectopic kidney. PMID- 21623134 TI - [Clinical characteristics and risk factors of reflux esophagitis: comparison with nonerosive reflux disease]. PMID- 21623135 TI - [Diagnosis and treatment of gastric MALT lymphoma]. AB - Gastric mucosa-associated lymphoid tissue (MALT) lymphoma represents approximately 40% of gastric lymphomas, and its incidence is increasing. An early diagnosis for gastric MALT lymphoma is important, but not easy due to non specific symptoms and endoscopic findings. Diagnosis is based on the histopathologic evaluation of multiple, deep and repeated biopsies taken from normal and any abnormal appearing sites of the stomach. In addition, the presence of Helicobacter pylori (H. pylori) infection must be determined to determine therapeutic approach. Endoscopic ultrasonography (EUS) is essential for the evaluation of regional lymph nodes and the depth of tumor invasion in the gastric wall, for predicting response to H. pylori eradication, and for monitoring tumor regression or recurrence. The eradication of H. pylori is recommended as an initial treatment for low-grade gastric MALT lymphoma with H. pylori infection. Both radiation therapy and chemotherapy are suitable alternative options for H. pylori-negative, refractory, or high-grade gastric MALT lymphoma. But, the role of surgery is diminishing. After treatment, strict endoscopic regular follow-up including EUS is recommended with multiple biopsies. However, controversy remains regarding the best diagnosis, treatment and follow-up strategy for this disease. PMID- 21623136 TI - [Clinical characteristics of gastroesophageal reflux disease with esophageal injury in korean: focusing on risk factors]. AB - BACKGROUND/AIMS: Recent studies suggest that the prevalence of gastroesophageal reflux disease (GERD) is increasing in Korea. However, studies on risk factors for GERD have yielded inconsistent results. The aims of this study were to compare clinical features between symptomatic syndromes without esophageal injury (=non-erosive disease [NED]) and syndromes with esophageal injury (=erosive disease [ED]), and to determine risk factors associated ED. METHODS: A total of 450 subjects who visited gastroenterology clinics of six training hospitals in Daegu from March 2008 to April 2010 were consecutively enrolled. The subjects were asked to complete a questionnaire which inquired about gastroesophageal reflux symptoms. The questionnaire also included questions about smoking, alcohol drinking, consumption of coffee, use of drugs, exercise, and other medical history. The subjects were subdivided into NED and ED groups. RESULTS: The proportion of subjects in each NED and ED group was 172 (38.2%) and 278 (61.8%). Male gender, smoking, alcohol drinking, consumption of coffee, large waist circumference, infrequent medication of antacids, aspirin and NSAIDs, infrequent and mild GERD symptoms were all significantly associated with ED on univariate analysis. Age, hiatal hernia, diabetes mellitus, body mass index, change in weight during 1 year, and number of typical GERD symptoms were not independent risk factors for ED. However, the association between ED and alcohol drinking, infrequent medication of antacids, mild typical GERD symptoms remained as strong risk factors after adjustments on multivariate logistic analysis. CONCLUSIONS: Independent risk factors associated with ED were alcohol drinking, infrequent medication of antacids and mild typical GERD symptoms. PMID- 21623137 TI - Characteristics of advanced gastric cancer undetected on gastroscopy. AB - BACKGROUND/AIMS: Stomach cancer can be easily diagnosed via endoscopy, but also possible to be missed. The aim of this study was to investigate the clinical and endoscopic characteristics of advanced gastric cancers that were not diagnosed based on endoscopic examination. METHODS: We evaluated patients who had newly diagnosed advanced gastric cancer that was undetected via endoscopy within the last six months. RESULTS: Sixteen patients were included in this study. The locations of the cancers were the cardia in six cases, the greater curvature side of the body in eight cases and the antrum in two cases. The histological findings were tubular type adenocarcinoma in 11 cases, with ten cases of moderately to poorly differentiated adenocarcinoma and five cases of signet ring cell type adenocarcinoma. CONCLUSIONS: Even advanced gastric cancer lesions may not be detected during endoscopy. If a patient continues to complain of upper gastrointestinal symptoms, even though endoscopy does not find abnormal findings, repeated endoscopy and/or additional diagnostic studies should be considered. PMID- 21623138 TI - [Importance of medication adherence to peginterferon-ribavirin combination therapy in patients with chronic hepatitis C]. AB - BACKGROUND/AIMS: The combination therapy with peginterferon and ribavirin is a standard treatment for patients with chronic hepatitis C. However, because of the long duration of the treatment and many complications, the reduction of adherence frequently occur. This study aimed to assess influences of reduced medication adherence in the combination therapy of chronic hepatitis C patients. METHODS: We retrospectively reviewed the medical records of 82 patients with chronic hepatitis C who received a combination therapy with peginterferon and ribavirin. The patients were categorized into 3 subgroups on the basis of medication adherence. Group 1 comprised patients who received >=80% of the recommended dosage of both peginterferon and ribavirin. Group 2 comprised those patients who received >=80% of the recommended dosage of only 1 drug. The patients of Group 3 received <80% of the recommended dosage of both the drugs. RESULTS: Sustained virologic response (SVR)s of patients in Group 1, 2 and 3 were 85.4% (41/48), 85.7% (18/21), and 38.5% (5/13), respectively (p=0.002). SVRs of genotype 1 patients in Group 1, 2 and 3 were 84.2% (16/19), 75% (9/12), and 14.3% (1/7) , respectively (p=0.003). SVRs of genotype non-1 patients in Group 1, 2 and 3 were 86.2% (25/29), 100% (9/9), and 66.7% (4/6), respectively (p=0.196). Furthermore are SVRs significantly differed with the degree of medication adherence to either peginterferon or ribavirin (p=0.003 and 0.021, respectively). In multivariate analysis, the peginterferon dose was a significant independent factor associated with SVR. CONCLUSIONS: Medication adherence of chronic hepatitis C patients to the combination therapy with peginterferon and ribavirin is very important for achieving SVR. In particular, we think that genotype 1 patients should maintain higher adherence than genotype non-1 patients. PMID- 21623139 TI - [Comparison among endoscopic variceal obliteration, endoscopic band ligation, and balloon-occluded retrograde transvenous obliteration for treatment of gastric variceal bleeding]. AB - BACKGROUND/AIMS: Endoscopic variceal obliteration (EVO), endoscopic variceal ligation (EVL), and balloon-occluded retrograde transvenous obliteration (BRTO) are used to manage gastric variceal bleeding. We compared the re-bleeding rates and survival times of these modalities. METHODS: The study enrolled 103 patients with suspected gastric variceal bleeding between July 2001 and May 2009. For the management of gastric variceal bleeding, 52 patients underwent EVO; 36, EVL; and 15, BRTO. We evaluated their laboratory results and vital signs, and calculated the Child score, Child classification, and Model for End-stage Liver Disease score. Rebleeding was defined as new-onset hematemesis, hematochezia, melena, or endoscopically proven bleeding. Time-to-rebleeding and survival time were examined by Kaplan-Meyer analysis. A value of p<0.05 indicated statistical significance. RESULTS: There were no significant differences in baseline characteristics among the three groups. The overall follow-up period averaged 65.13 months. During follow-up, rebleeding occurred in 17 patients (11 EVO, 5 EVL, and 1 BRTO). The times-to-rebleeding were 63.59, 75.79, and 51.41 months for EVO, EVL, and BRTO, respectively, and did not differ significantly (p=0.515). The median survival times were 77.42, 70.14, and 42.79 months, respectively, and also were not different significantly (p=0.978). CONCLUSIONS: There were no significant differences in the time-to-rebleeding or survival time among EVO, EVL, and BRTO. Further prospective, large-scale studies are needed. PMID- 21623140 TI - [A case of acute phlegmonous gastritis causing gastroparesis and cured with medical treatment alone]. AB - Acute phlegmonous gastritis is an uncommon disease, often fatal condition characterized by suppurative bacterial infection of the gastric wall. It has a high mortality rate mainly because the diagnosis is usually made late. Until recently, gastrectomy in combination with antibiotics was recommended. We had experienced a case of 66-year-old man presented with epigastric pain, nausea, vomiting, and hematemesis, followed by aspiration pneumonia. At upper gastrointestinal endoscopy, the gastric lumen was narrow, and the mucosa was severely inflamed, which was erythematous, swelled, and showed necrotic areas covered with purulent exudate. Klebsiella oxytoca and Acinetobacter lwoffii were isolated in the gastric tissue culture. Contrast-enhanced computerized tomography scan of abdomen demonstrated diffuse gastric wall thickening and an intramural abscess in the gastric antral wall. Although delayed gastric emptying by gastroparesis prolonged the in-hospital period, the only medical treatment with antibiotics alone successfully cured the patient without gastrectomy. PMID- 21623141 TI - [A case of autoimmune hepatitis following acute hepatitis A]. AB - The pathogenesis of autoimmune hepatitis (AIH) is unclear, but viral infections have been proposed as a potential trigger in patients with genetic predisposition. We report a case of AIH following acute hepatitis A (AHA). A 57 year-old woman presented with fatigue and pitting edema for last 3 months. She had been diagnosed as an AHA 15 months ago based on clinical features, biochemical tests and positive HAV IgM antibody at a local clinic. Her biochemical tests was normalized one month after AHA diagnosis, but the serum levels of aminotransferase started to rise four months after AHA diagnosis. Antinuclear antibody was positive at a titer of 1:40, and anti-smooth muscle antibody was also positive. Hypergammaglobulinemia and liver pathology were typical for AIH. The patients had a score of 17 according to the International Autoimmune Hepatitis Group's system. She was given prednisolone and azathioprine and showed complete response to immunosuppressive therapy. The present case is the first report on AIH triggered by AHA in Korea. PMID- 21623142 TI - [A case of primary mucoepidermoid carcinoma arising from the common bile duct]. AB - Mucoepidermoid carcinoma of the bile duct is an extremely rare tumor. Seventeen cases originating from intrahepatic bile duct and 2 cases from common hepatic duct have been reported in the English literature. Mucoepidermoid carcinoma arising from the common bile duct has not been previously reported. A 68 year-old man was admitted due to obstructive jaundice. Computed tomography showed a malignant tumor of the common bile duct located in the intrapancreatic segment. Filling defects of the distal common bile duct was seen on endoscopic retrograde cholangiogram. Under the impression of bile duct cancer, pylorus-preserving pancreatoduodenectomy was performed. Histologic diagnosis of the resected specimen was mucoepidermoid carcinoma of the common bile duct. After surgery, the patient received concurrent chemoradiotherapy, and planned to receive additional chemotherapy. We herein report on a first case of primary mucoepidermoid carcinoma of the common bile duct, and review the literature. PMID- 21623143 TI - [A case of a retroperitoneal schwannoma presenting as hypermetabolic mass in PET CT]. AB - Schwannoma is a benign neoplasm of the Schwann cells of the neural sheath. Most schwannomas occur in the head and neck, and extremities and rarely in the retroperitoneal space. The differentiation of a schwannoma from other malignant tumor or benign tumor is very difficult on a preoperative examination with ultrasonography, computed tomography or magnetic resonance imaging. Furthermore, the lesion with increased fluorodeoxyglucose uptake in PET-CT cannot exclude malignant tumor. Therefore, this lesion needs surgical excision and a histological examination with immunohistochemical staining. We report a case of schwannoma occuring in the retroperitoneal space that incidentally discovered by PET-CT for health-check up. Pathologic confirmation by laparoscopic excision was done. PMID- 21623144 TI - [Gastric wall abscess]. PMID- 21623146 TI - A case of Behcet's disease with massive lower gastrointestinal bleeding. PMID- 21623147 TI - Glucocorticoid-induced osteoporosis in men. AB - Osteoporosis and fractures are a common consequence of glucocorticoid therapy for inflammatory disorders. Men fracture approximately 10 yr later in life than women and receive less attention as regards osteoporosis risk, including in glucocorticoid-induced osteoporosis (GIOP). In addition, while men are less likely to have certain rheumatologic disorders often treated with glucocorticoids, men are more likely to have chronic obstructive pulmonary disease, inflammatory bowel disease, and organ transplantation as reasons for use of oral glucocorticoids. Attempts to improve recognition of GIOP in general have not been successful, and since men are considered less at risk for osteoporosis in general, attention to men with GIOP is even less. Evaluation of GIOP is similar in men and women, and most modern treatment studies of GIOP have included men. Thus, alendronate, risedronate, and zoledronic acid are Food and Drug Administration (FDA)-approved bisphosphonates for GIOP in men. Teriparatide is also FDA-approved for GIOP. In one 36-month trial of teriparatide vs alendronate for GIOP in men and women, the anabolic agent led to a greater increase in bone density and was associated with a lower incidence of morphologic vertebral fractures. Thus, while good management is available for GIOP, recognition of men at risk is the most important step in improving outcomes. PMID- 21623148 TI - A comparative study of pre-operative imaging methods in patients with primary hyperparathyroidism: ultrasonography, 99mTc sestamibi, single photon emission computed tomography, and magnetic resonance imaging. AB - AIM: This study compares the accuracy rates achieved in ultrasonography (US), 99mTc-sestamibi (MIBI), single photon emission computed tomography (SPECT), and magnetic resonance imaging (MRI) as imaging methods used in the pre-operative localization of the enlarged parathyroid glands. SUBJECTS AND METHODS: For the purposes of this study, US, MIBI, SPECT, and MRI were performed on 98 patients with primary hyperparathyroidism (pHPT). All patients underwent parathyroidectomy. RESULTS: Pre-operative localization of an abnormal parathyroid gland was successfully performed in 82 of the cases scanned with US (83.7%), while the result was 66 in the cases scanned with MIBI (67.3%), 71 of the cases were successfully localized with SPECT (72.4%), while MRI revealed the diseased gland in only 60 of the total cases (61.2%). In MIBI-positive and -negative patients there was a statistically significant difference among cases in terms of adenoma volume (1.30+/-1.51 vs 0.58+/-0.91, p<0.05). Sensitivity, specificity and diagnostic accuracy values were 87.2%, 25.0%, and 83.0%; 70.2%, 50.0%, and 69.4%; 75.5%, 50.0%, and 74.5%; 63.8%, 50.0%, and 63.3% for US, MIBI, SPECT, and MRI, respectively. The respective values for sensitivity, specificity, and diagnostic accuracy were 94.9%, 25.0%, and 91.1% when US was combined with MIBI. CONCLUSIONS: Combining US and MIBI as imaging methods for pre-operative imaging of pHPT often produces more satisfactory results. While the accuracy of US is relatively low in the ectopic localizations, the size of the lesion can be an important factor in the accuracy achieved with MIBI. PMID- 21623149 TI - Acylated and unacylated ghrelin levels in normal weight and obese children: influence of puberty and relationship with insulin, leptin and adiponectin levels. AB - BACKGROUND: Ghrelin circulates in blood as acylated (AG) and unacylated (UAG) ghrelin. The physiological role of the two forms is poorly understood, in particular in childhood. Aim of the study was to evaluate the AG and UAG levels in obese and normal weight (NW) children, pre-pubertal and pubertal, and their relationship with insulin, leptin and adiponectin levels. SUBJECTS AND METHODS: A population based study in which AG, UAG, leptin, adiponectin, glucose, insulin, testosterone or estradiol levels, insulinemic indexes were evaluated in 82 NW and 58 obese (OB) children. RESULTS: Both ghrelin forms in NW were higher (AG, p<0.02; UAG, p<0.0001) than in OB subjects, with similar ratio AG/UAG . While no differences were observed for gender, puberty AG (p<0.01) and UAG (p<0.0001) levels were higher in pre-pubertal than pubertal NW and OB subjects. Adiponectin levels in NW subjects were higher (p<0.001), while leptin and insulin levels were lower (p<0.0001) than in OB subjects. NW children showed homeostasis model assessment (HOMA) and HOMAbeta indices lower than OB children (p<0.0001) with a higher a quantitative insulin sensitivity check index (p<0.0001). AG and UAG levels correlated to each other (p<0.0001), each showing a negative correlation to age, height, weight and body mass index. Both forms, but more strongly UAG, correlated with adiponectin, leptin, and insulin. CONCLUSIONS: OB children show lower levels of both AG and UAG when compared to NW subjects, with lower levels during puberty. These results demonstrate a peculiar strong relationship between UAG levels and metabolic parameters in the pediatric population, suggesting a role for UAG in metabolic functions. PMID- 21623150 TI - Lipid profile and nutritional intake in children and adolescents with Type 1 diabetes improve after a structured dietician training to a Mediterranean-style diet. AB - AIM: To evaluate if nutritional intakes and lipid profile fulfill international guidelines and recommendations before and after a structured dietician training to a Mediterranean- style diet in an Italian pediatric population with Type 1 diabetes. METHODS: A 6-month prospective cohort study. Baseline and after intervention nutritional intakes, lipid profile, glycated hemoglobin (HbA(1c)), and clinical parameters of 96 children and adolescents with Type 1 diabetes were assessed. A comparative computerized system which was approved and validated by the Italian Diabetologist Association was used to define the amounts of nutrients. RESULTS: At baseline mean daily dietary intakes of carbohydrates, proteins, and lipids were respectively (mean +/- SEM) 51.8 +/- 0.5, 15.9 +/- 0.2, 33.8 +/- 0.6%, with a contribution of cholesterol of 248.7 +/- 12.5 mg/day. Fiber assumption was 18.0 +/- 0.4 g/day. The 64.5% and 29.1% (p<0.0001) of subjects had at least one lipid parameter higher than 75(th) and 95(th) percentiles, respectively, of selected cut points (American Diabetes Association guidelines for total and LDL-cholesterol and American Academy of Pediatrics standards for HDL-cholesterol and triglycerides). Six months after the dietician intervention, dietary lipids and cholesterol decreased (p<0.0001) while fibers (p<0.0001) increased. LDL-cholesterol, non-HDL-cholesterol, and total cholesterol:HDL cholesterol ratios significantly decreased (p<0.001) with a reduction of rate of subjects with at least one pathological lipid parameter (p<0.01) independently by weight and glucose control. CONCLUSIONS: Italian pediatric subjects with Type 1 diabetes present a balanced diet with exception of lipids intake and a suboptimal lipid profile. A structured dietician training to a Mediterranean-style diet improves the quality of nutrient intakes being followed by a reduction of LDL cholesterol, non- HDL-cholesterol, and total cholesterol:HDL-cholesterol ratios. PMID- 21623151 TI - Treatment with somatostatin analogs and chemoembolization of liver metastases for severe hypoglycemia in malignant insulinomas. AB - BACKGROUND: Preventing hypoglycemia is of vital importance and a major challenge in patients with severe symptomatic hypoglycemia related to malignant unresectable insulinomas, but there is no consensus treatment. PATIENTS: Five patients with malignant unresectable insulinomas were referred to our department for severe hypoglycemia. At referral the five patients were dependent on iv infusion of glucose solution. Patient 1 had a locally invasive 5-cm insulinoma, patients 2, 3 and 4 had multiple liver metastases and patient 5 had a 2.5-cm pancreatic tumor with multiple liver and lung metastases. Before referral to our department, 4/5 patients had been administered systemic chemotherapy and 3/5 therapeutic doses of radiolabeled octreotide without any benefit on blood glucose levels. Octreoscan scintigraphy was positive in 4 patients (patients 1-4). Diazoxide alone or combined with glucocorticoids had failed to control hypoglycemia. Continuous sc administration of octreotide (up to 1500 MUg/day) resulted in normalization of blood glucose levels in patient 1. Chemoembolization of liver metastases normalized blood glucose levels in patient 2, minimized hypoglycemia in patients 3 and 4, and normalized blood glucose levels in patient 5 when followed by subcutaneous administration of octreotide (2000 MUg/day). Chemoembolization had to be repeated four times in patient 3 to control blood glucose levels. CONCLUSION: Chemoembolization of liver metastases and high-dose octreotide in responsive patients (alone or combined with chemoembolization) can control severe hypoglycemia in patients with symptomatic malignant unresectable insulinomas; the efficacy of octreotide can be improved after chemoembolization of liver metastases. PMID- 21623152 TI - Carotid intima-media thickness in Chinese Type 2 diabetic subjects with or without microalbuminuria. AB - BACKGROUND: To examine the association of microalbuminuria (MAU) with the carotid intima-media thickness (CIMT) in Chinese Type 2 diabetic subjects. MATERIALS AND METHODS: Two hundred and thirty-nine patients (64+/-13 yr, 154 males) were divided into 2 groups: one with MAU (no.=119) or one without (no.=120). We recorded clinical and biochemical data as well as CIMT and ankle-brachial index (ABI). RESULTS: The patients with MAU had had diabetes mellitus (DM) longer, had higher blood pressure (BP). They also had lower estimated glomerular filtration rate (eGFR) and higher levels of circulating glucose, glycated hemoglobin, high sensitivity C-reactive protein than those without. Lower mean ABI was found in those with MAU, however, they did not have higher mean CIMT (0.72+/-0.15 vs 0.71+/-0.16 mm, p=0.525). In patients without MAU, CIMT correlated with age, DM duration, systolic BP, eGFR, albumin- to-creatinine ratio, and ABI. However, in those with MAU, CIMT correlated only with age and eGFR. Multivariate regression analysis revealed that mean CIMT correlated only with age for patients without MAU, but correlated with age and body mass index for those with MAU. Dividing the patients into 5 age groups, we found that the older the patient, the higher the mean CIMT with no group differences between those with and without MAU in both genders. However, patients with eGFR below 60 ml/min/1.73 m(2) had higher mean CIMT than those above (0.75+/-0.16 vs 0.69+/-0.14 mm, p=0.005). CONCLUSIONS: Type 2 diabetic patients with MAU were not associated with higher CIMT. Conversely, those with deterioration of renal function were more likely associated. PMID- 21623153 TI - Pituitary autoimmunity is associated with hypopituitarism in patients with primary empty sella. AB - OBJECTIVE: Some evidence suggests that late stage autoimmune hypophysitis (AH) may result in empty sella (ES). Aim of the study was to assess the prevalence of serum pituitary antibodies (PitAb) and their correlation with pituitary function in patients with ES. DESIGN: In this casecontrol study 85 patients with primary ES, 16 patients with ES secondary to head trauma, 214 healthy controls, and 16 AH were enrolled in a tertiary referral center. METHODS: PitAb were assessed in all cases and controls. Endocrine function was assessed by basal hormone measurement and dynamic testing in all ES cases. RESULTS: PitAb prevalence was higher in primary ES (6%) than in healthy subjects (0.5% p=0.003) and lower than in AH patients (50%, p<0.0001). PitAb were not found in patients with secondary ES. Hypopituitarism was found in 49% of primary ES and in 62% of secondary ES (p=0.34). A positive correlation between the presence of PitAb and hypopituitarism was found in primary ES (p=0.02). CONCLUSIONS: The significant association between pituitary autoimmunity and hypopituitarism suggests that ES, in selected cases, could be the final result of AH. PMID- 21623155 TI - Continuous insulin infusion is associated with a reduced post-surgical length of stay, but not with the complication rate, in patients with diabetes mellitus undergoing coronary artery bypass graft. AB - OBJECTIVE: To establish if glucose management with continuous intravenous insulin infusion (CII) in the early post-operative period after coronary artery bypass graft (CABG) surgery is associated with complication rate and length of hospital stay (LOS) in patients with diabetes mellitus (DM). RESEARCH DESIGN AND METHODS: We reviewed the records of 587 patients with DM who underwent CABG from January 1999 until January 2008; 316 patients were placed on CII, while 271 patients were treated with subcutaneous insulin. We examined patient age, glycated hemoglobin (HgbA1c), 24- and 72-h post-operative average capillary blood glucose (CBG), length of stay (LOS), and the rate of complications. RESULTS: There was no difference in HgbA1c between the groups. Mean CBG values at both 24 h and 72 h remained the same in the CII group (167 mg/dl), while in the non-CII group they were 194 mg/dl and 189 mg/dl, respectively (p<0.001 between the groups). Post surgical median LOS was 6 days in the CII group and 6.5 days in the non-CII group (p=0.003). Complications occurred at similar rate (in 10% and 11% of patients) in the two groups. CONCLUSIONS: CII is associated with a reduced post-surgical LOS in patients with DM who undergo CABG. PMID- 21623154 TI - Iodothyronine deiodinases and cancer. AB - Thyroid hormones (TH) regulate key cellular processes, including proliferation, differentiation, and apoptosis in virtually all human cells. Disturbances in TH pathway and the resulting deregulation of these processes have been linked with neoplasia. The concentrations of TH in peripheral tissues are regulated via the activity of iodothyronine deiodinases. There are 3 types of these enzymes: type 1 and type 2 deiodinases are involved in TH activation while type 3 deiodinase inactivates TH. Expression and activity of iodothyronine deiodinases are disturbed in different types of neoplasia. According to the limited number of studies in cancer cell lines and mouse models changes in intratumoral and extratumoral T3 concentrations may influence proliferation rate and metastatic progression. Recent findings showing that increased expression of type 3 deiodinases may lead to enhanced tumoral proliferation support the idea that deiodinating enzymes have the potential to influence cancer progression. This review summarizes the observations of impaired expression and activity in different cancer types, published to date, and the mechanisms behind these alterations, including impaired regulation via TH receptors, transforming growth factor-beta, and Sonic-hedgehog pathway. Possible roles of deiodinases as cancer markers and potential modulators of tumor progression are also discussed. PMID- 21623156 TI - The natural history of the hyperthyrotropinemia of children born prematurely. AB - OBJECTIVE: Non-autoimmune hyperthyrotropinemia has been previously reported among children born prematurely. The aim of this study was to follow up their thyroid function, volume, and structure and to investigate the relationship with growth, IGF-I, lipid profile, and insulin sensitivity. METHODS: Seventy-two children born prematurely (33.2+/-2.2 weeks), 26 appropriate (AGA) and 46 small for gestational age (SGA), were evaluated at the age of 7.6+/-2.3 yr (time 1) and at the age of 11.4+/-2.3 yr (time 2). We also measured TSH, free T(3) (fT(3)), free T(4) (fT(4)), thyroperoxidase antibodies (TPO-Ab), thyroglobulin antibodies (TG-Ab), thyroid ultrasound, auxological parameters, lipid profile, glucose, and insulin level. RESULTS: In the AGA group TSH was similar in both times (2.7+/-1.0 vs 3.0+/-0.9 mU/l) and above the upper normal limit in 4 (15.4%) subjects at time 1 and in 6 (23.7%) subjects at time 2 (ns). In the SGA group, TSH was similar in both times (2.8+/-1.2 vs 2.5+/-1.0 mU/l) and above the upper normal limit in 11 (23.9%) subjects at time 1 and 5 (10.8%) subjects at time 2 (ns). fT(4) and fT(3) were always normal and TPO- and TG-Ab absent. Thyroid volume increased progressively, but significantly only in the AGA group (p=0.0005). The thyroid structure was always normal and there was no influence on the growth and the biochemical profile. CONCLUSIONS: Some ex-premature babies show a mild and variable thyroid dysfunction, which does not seem to evolve toward an overt thyroid dysfunction. PMID- 21623157 TI - Changes in lifestyle improve body composition, thyroid function, and structure in obese children. AB - BACKGROUND: Alterations in thyroid function and structure have been reported in obesity. Function reverses to normal after weight loss, but nothing is known about structure. AIM: To evaluate the effect of weight loss on thyroid function and structure in obese children. SUBJECTS AND METHODS: The study was conducted in 72 overweight and obese children. Measurement of free T(3) (fT(3)), free T4 (fT(4)), TSH, antithyroid- antibodies and a thyroid ultrasound was performed at the beginning (phase 1) and after a period of 1.8+/-1.0 yr of lifestyle intervention (phase 2). RESULTS: Height SD score (SDS), body mass index SDS, total fat mass did not change from phase 1 to phase 2. Percentage of fat free mass decreased significantly (p<0.05). Waist/height ratio decreased (0.6+/-0.1 vs 0.5+/-0.1; p<0.05) as well as waist/hip ratio (0.9+/-0.1 vs 0.8+/-0.1; p<0.05). In phase 1, TSH was 2.8+/-1.7 mU/l; in phase 2, it was 2.2+/-0.9 mU/l (p<0.05); 17.2% of children showed a TSH level above the normal range (3.6 mU/l) in phase 1, and 6.2% in phase 2 (p<0.05). fT(4) was 10.8+/-2.2 pg/ml in phase 1 and 10.7+/ 1.9 pg/ml in phase 2. fT(3) was 4.4+/-1.3 pg/ml (phase 1) and 3.9+/-1.1 pg/ml (phase 2) (p<0.05). Thyroid volume was -0.5+/-0.8 SDS (phase 1) and -0.5+/-1 SDS (phase 2). A non-significant improvement in thyroid structure was observed. CONCLUSIONS: In conclusion, healthier lifestyle improves body composition, thyroid function, and structure. PMID- 21623158 TI - Addition of chlorine during water purification reduces iodine content of drinking water and contributes to iodine deficiency. AB - Drinking water is the major natural source of iodine in many European countries. In the present study, we examined possible sites of iodine loss during the usual water purification process.Water samples from 6 sites during the technological process were taken and analyzed for iodine content. Under laboratory circumstances, prepared iodine in water solution has been used as a model to test the effect of the presence of chlorine. Samples from the purification sites revealed that in the presence of chlorine there is a progressive loss of iodine from the water. In the chlorine concentrations employed in the purification process, 24-h chlorine exposure eliminated more than 50% of iodine when the initial iodine concentration was 250 MUg/l or less. Iodine was completely eliminated if the starting concentration was 16 MUg/l.We conclude that chlorine used during water purification may be a major contributor to iodine deficiency in European communities. PMID- 21623159 TI - MicroRNAs and lung cancer: from markers to targets. PMID- 21623160 TI - Microparticulate systems for targeted drug delivery to phagocytes. PMID- 21623161 TI - An intimate connection between ubiquitination and compartmentalized cAMP signaling. PMID- 21623162 TI - Kinetic profiling of the c-Myc transcriptome and bioinformatic analysis of repressed gene promoters. AB - Mammalian c-Myc is a member of a small family of three related proto-oncogenic transcription factors. c-Myc has an unusually broad array of regulatory functions, which include roles in cell cycle and apoptosis, a variety of metabolic functions, cell differentiation, senescence, and stem cell maintenance. c-Myc modulates the expression of a very large number of genes, but the magnitude of the majority of the regulatory effects is only 2-fold or less. c-Myc can both activate and repress the promoters of its target genes. Identification of genes directly regulated by c-Myc has been an enduring question in the field. We report here microarray expression profiling of a high resolution time course of c-Myc induction, using fibroblast cells in which c-Myc activity can be modulated from null to physiological. The c-Myc transcriptome dataset presented is the largest reported to date with 4,186 differentially regulated genes (1,826 upregulated, 2,360 downregulated, 1% FDR). The gene expression patterns fit well with the known biological functions of c-Myc. We describe several novel findings and present tools for further data mining. Although the mechanisms of transcriptional activation by c-Myc are well understood, how c-Myc represses an even greater number of genes remains incompletely described. One mechanism involves the binding of c-Myc to other, positively acting transcription factors, and interfering with their activities. We identified rapid-response genes likely to be direct c-Myc targets, and analyzed the promoters of the repressed genes to identify transcription factors that could be targets of c-Myc repression. PMID- 21623163 TI - Villin1, a diagnostic marker for endometrial adenocarcinoma with high grade nuclear atypia. AB - Villin1 (VIL1) has a role in regulating actin dynamics, cell morphology, anti apoptotic mechanisms, and epithelial-to-mesenchymal transition. Previously we reported VIL1 as a novel diagnostic marker for cervical adenocarcinoma (AC) with poor radioresponse. This study further investigated the diagnostic role of VIL1 in gynecological tumors especially endometrial AC. We recruited 107 patients with AC (41 tumors in the corpus and 66 tumors in cervix), most of whom treated by total abdominal hysterectomy. Immunohistochemical analysis revealed VIL1-positive tumors in 37% of cases; 10 of 41 corpus tumors and 30 of 66 tumors in the cervix. VIL1-positive tumors were further examined histologically and immunostained for epithelial cell surface marker, EpCAM, and mesenchymal stem cell marker, CD44. Most of these tumors were CD44 negative and EpCAM positive, and the cytoplasmic VIL1 immunoreactivity in endometrial AC was more selective than EpCAM in reflecting histological aggressiveness with high grade nuclear atypia. This study confirmed our previous finding of VIL1 as a diagnostic marker of cervical AC. In addition, VIL1 immunostaining was detected in 25% of endometrial AC cases. These results suggested the existence of an aggressive and VIL1-positive subtype of gynecological tumor. PMID- 21623164 TI - Targeting the inflammatory pathways to enhance chemotherapy of cancer. AB - Epidemiological and experimental evidence has emerged that a dysregulated inflammation is associated with most of the tumors. Recent studies have begun to unravel molecular pathways linking inflammation and cancer. The identification of transcription factors such as NF-kappaB, STAT3, HIF-1 alpha and their gene products such as COX-2, cytokines, chemokines and chemokine receptors have laid molecular foundation for the decisive role of inflammation in carcinogenesis. Inflammation contributes to survival and proliferation of malignant cells, tumor angiogenesis, metastasis and reduced response to chemotherapy. In view of their involvement at different stages of tumor development, inflammatory pathways represent attractive targets for cancer prevention and therapy. However, advances in this field have not been fully realized, and challenge remains to obtain clinical data from patients and then to utilize this information for optimal and personalized therapy. We present evidence that targeting inflammatory pathways have a potential role to improve chemotherapy of cancer. PMID- 21623166 TI - Inhibition of N-myc downstream-regulated gene 2 in prostatic carcinoma. AB - To study the expression of N-myc Downstream Regulated Gene-2 (NDRG2) in prostatic carcinoma (PCA) tissue and in different PCA cell lines, and to investigate its clinical and pathological implications, 144 PCA and benign prostatic hyperplasia (BPH) tissue sections were analyzed retrospectively with immunohistochemistry (S P method). The expression levels of NDRG2 and c-Myc in prostate cell lines were detected through Western blot. The effects of adenovirus-mediated NDRG2 on PC3 cells and PC3 nude mouse xenografts was observed through cell growth curves, tumor growth curves, flow cytometry (FCM), transmission electron microscopy (TEM) and TUNEL staining. The NDRG2 gene was highly expressed in BPH tissues, but not in carcinomatous ones (chi(2)=25.98, p < 0.001). Furthermore, positive expression of NDRG2 was negatively correlated with the Gleason score (r = -0.445, p< 0.001) and the c-myc level (r = -0.311, p < 0.001). However, positive expression of NDRG2 was not correlated with pTNM tumor stages or the serum concentration of prostate-specific antigen (PSA) (p > 0.05). The expression of the NDRG2 genes was low in the three PCA cell lines. PC3 cells infected by pAD-cmv-NDRG2 showed inhibition of proliferation both in vitro and vivo. To sum up, NDRG2 may be involved in the carcinogenesis and progression of PCA. Moreover, adenovirus mediated NDRG2 can suppress the proliferation of PC3 cells significantly both in vitro and in vivo. These results indicate that NDRG2 may become a new target gene for PCA diagnosis and therapy. PMID- 21623167 TI - Humanized mice for Salmonella typhi infection: new tools for an old problem. AB - Salmonella enterica serovar Typhi (S. typhi), is a human restricted pathogen and the causal agent of typhoid fever. Although the use of antimicrobial drugs or vaccines has served as an effective therapeutics strategy against typhoid fever, the recent surge in multidrug resistant strains of S. typhi presents a major health concern worldwide. Progress on typhoid research has been limited in the past due to the lack of a suitable animal model that recapitulates the hallmark immunological features of human typhoid fever. We have recently developed a humanized immune system (HIS) mouse model that after intravenous challenge with S. typhi displayed classical manifestations of human typhoid fever including meningitis, liver pathology and mortality. Concurrent to our studies, two other groups also have developed humanized mouse models of S. typhi infections employing different protocols. All these recently adopted animal models of S. typhi infections provide promise for a new era of S. typhi research that may expedite detailed understanding of the cellular and molecular mechanisms of this bacterial infection and investigations of new antimicrobials and vaccines for effective control. PMID- 21623168 TI - Effectors of Salmonella pathogenicity island 2: An island crucial to the life of Salmonella. AB - The tug of war between a pathogen and its host has been one of the most amazing stories in the field of microbial pathogenesis for ages. The strongest known species of all living organisms is the Homo sapiens and yet it is incredible how a pathogen of the size of few microns is smart enough to defeat this mightiest group of survivors. It is of utmost interest to understand the mechanisms behind the successful habitation of a pathogen inside the ever-resisting and complicate human body. Numerous examples of diseases caused by such pathogens exist which intrigues us to venture in the world of host-pathogen interactions. PMID- 21623169 TI - beta-Cell replication and islet neogenesis following partial pancreatectomy. AB - Partial pancreatectomy is one of the most commonly used models in the study of beta-cell regeneration. The mechanism by which regeneration occurs in this model has been controversial, with some claiming that islet and beta-cell neogenesis is important, while others claim that beta-cell replication is predominant. Here, we combined a time course analysis with continuous BrdU administration to study beta cell regeneration following partial pancreatectomy. While exocrine cells in regenerating areas were highly proliferative and positive for BrdU, islets in regenerating areas were negative for BrdU one week after partial pancreatectomy, suggesting that they were derived from preexisting islets rather than being neogenic. The insulin-positive cells in ducts that have been reported by others and taken as evidence of beta-cell neogenesis were present in regenerating regions of the pancreas, but were relatively uncommon and were not highly proliferative, suggesting that they could not account for significant islet neogenesis. Consistent with a lack of islet neogenesis, regenerating areas following a second partial pancreatectomy were devoid of islets. beta-cell replication was detectable at a high frequency two weeks following partial pancreatectomy and was present at a similar frequency in both regenerating and preexisting regions of the pancreas. In summary, our data indicate that islet neogenesis following partial pancreatectomy does not occur. PMID- 21623170 TI - MRE11 promotes AKT phosphorylation in direct response to DNA double-strand breaks. AB - AKT is hyper-activated in many human cancers and promotes proliferation and cancer cell survival in response to DNA damaging agents. Ionizing radiation (IR) produces DNA double strand breaks (DSB) and activates AKT, however a direct mechanism linking intra-nuclear DSB and AKT signaling is lacking. Here we demonstrate that AKT is phosphorylated following IR in benign and malignant cells and, using colony-forming assays and in vitro rejoining assays, show that AKT promotes non-homologous end joining-mediated DSB repair and cell survival following IR. Further studies revealed that pAKT-S473, but not pAKT-T308 or total AKT, accumulates in the vicinity of IR-induced DSB and co-localizes with gammaH2AX and ATM-pSer1981. Based on whole-cell IR, nuclear UV microbeam, and endonuclease-induced DSB studies, we observed that pAKT-S473 is up-regulated by a DSB-induced signaling cascade, and this is dependent on the DSB sensor protein, MRE11. MRE11-dependent pAKT-S473 did not require the MRE11 endonuclease domain. The histone ubiquitin ligase RNF168 is also required for DSB-induced pAKT-S473, and DSB-induced pAKT-S473 is independent of DNA-PKcs, PI3K, and ATR. These data demonstrate that DSB activate a signaling cascade that directly promotes a PI3K independent pathway of AKT phosphorylation that is dependent on MRE11-ATM-RNF168 signaling. Thus, these data directly link the presence of DNA breaks to AKT mediated cell survival and support AKT as a target for cancer therapy. PMID- 21623171 TI - Inhibition of Aurora-A kinase induces cell cycle arrest in epithelial ovarian cancer stem cells by affecting NFKB pathway. AB - Recurrent ovarian cancer is resistant to conventional chemotherapy. A sub population of ovarian cancer cells, the epithelial ovarian cancer stem cells (EOC stem cells) have stemness properties, constitutive NFkappaB activity, and represent the chemoresistant population. Currently, there is no effective treatment that targets these cells. Aurora-A kinase (Aurora-A) is associated with tumor initiation and progression and is overexpressed in numerous malignancies. The aim of this study is to determine the effect of Aurora-A inhibition in EOC stem cells. EOC stem cells were treated with the Aurora-A inhibitor, MK-5108. Cell growth was monitored by Incucyte real-time imaging system, cell viability was measured using the Celltiter 96 assay and cytokine levels were quantified using xMAP technology. The intracellular changes associated with MK-5108 treatment are: (1) polyploidy and cell cycle arrest; (2) inhibition of NFkappaB activity; (3) decreased cytokine production; and (4) nuclear accumulation of IkappaBalpha. Thus, inhibition of Aurora-A decreases cell proliferation in the EOC stem cells by inducing cell cycle arrest and affecting the NFkappaB pathway. As EOC stem cells represent a source of recurrence and chemoresistance, these results suggest that Aurora-A inhibition may effectively target the cancer stem cell population in ovarian cancer. PMID- 21623172 TI - Glucagon plays an important role in the modification of insulin secretion by leptin. AB - Obese people show marked hyerinsulinemia, but the exact mechanism has not been clarified. Hyperleptinemia is one of possible candidates, although there is an obvious difference in the effect of leptin on insulin secretion between isolated pancreatic islets and beta-cell line. Since glucagon may modulate the effect of leptin on insulin secretion, we determined the influences of glucagon in the leptin effect on insulin secretion. The influences of glucagon in the leptin effect on insulin secretion for 10 minutes were determined by using isolated mouse islets and HIT-T 15 cells. The influences of 3-isobutyl-1- methylxanthine (IBMX), forskolin, and dibutyryl cyclic AMP were investigated in the leptin effect on insulin secretion. Leptin-inhibited insulin and glucagon secretion in isolated mouse pancreatic islets. In contrast, leptin stimulated insulin secretion in isolated mouse islets previously incubated with monoclonal anti glucagon antibodies for 18 hours. In HIT-T 15 cells, leptin dose-dependently increased insulin secretion, but this effect was attenuated by the addition of glucagon. The stimulatory effect of leptin on insulin secretion was attenuated by 48 hour pre-incubation with glucagon. In the presence of 100 mM IBMX, leptin decreased insulin secretion from HIT-T 15 cells. Leptin also reduced insulin secretion in the presence of 1mM forskolin or 1mM dibutyryl cyclic AMP. The leptin effects on insulin secretion were affected by the existence of glucagon. Intracellular cyclic AMP concentrations may determine the leptin effects on insulin secretion in pancreatic beta-cells. PMID- 21623173 TI - Electrophysiological identification of mouse islet alpha-cells: from isolated single alpha-cells to in situ assessment within pancreas slices. AB - Investigation of alpha-cells has long been constrained by their scarce population and localization at the islet mantle which exposes alpha-cells to injury by conventional islet isolation and dispersion to single cells that employ damaging enzymatic and mechanical means. To surmount these limitations, we recently reported employing the pancreas slice preparation which enables highly efficient acute in situ electrophysiological (patch clamp) examination of alpha-cells within its unperturbed native social environment with preserved paracrine regulation. In this review, we compare the electrophysiological properties of alpha-cells in these three preparations, and discuss the current view of glucose regulation of alpha-cells. We discuss current genetic mouse models that flurophore-tagged alpha-cells (GYY) and beta-cells (MIP-GFP) which can reliably identify islet cells to facilitate their study. Combining these strategies should enable future studies directed at the precise assessment of the perturbation in intrinsic and paracrine regulation of alpha-cells contributing to abnormal glucose homeostasis in diabetes. PMID- 21623175 TI - Child and adolescent psychiatry. Current world literature. PMID- 21623177 TI - Pleural effusions occurring with right heart failure. AB - PURPOSE OF REVIEW: Pleural effusions commonly occur in patients with left heart failure. However, there is increasing evidence that patients with pulmonary hypertension and isolated right heart failure frequently have pleural effusions. RECENT FINDINGS: Three recent studies have evaluated the incidence of pleural effusions without an alternate explanation in patients with idiopathic/familial pulmonary arterial hypertension (14%), pulmonary arterial hypertension associated with connective tissue diseases (33%), and portopulmonary hypertension (30%). The majority of patients in all three studies with pleural effusions without an alternate explanation were found to have isolated right heart failure. In these studies, mean right atrial pressures and death during follow-up were significantly higher in patients with pleural effusions and isolated right heart failure compared to patients with no pleural effusions. SUMMARY: Pleural effusions without an alternate explanation occur commonly in at least three subtypes of pulmonary arterial hypertension. The majority of patients with pleural effusions also have isolated right heart failure that is thought to be responsible for the development of the effusions. Patients presenting with pulmonary hypertension should be evaluated for pleural effusions, and if present, should receive a work-up for right heart failure. PMID- 21623179 TI - New research highlights the role of patient safety culture and safer care. PMID- 21623180 TI - Magnet learning communities. PMID- 21623181 TI - Documentation of contact precautions in an electronic health record. AB - Contact precautions are implemented to reduce transmission of multidrug-resistant organisms but may also increase hospital costs and patient complications. The goal of this study was to determine the prevalence of documentation of contact precautions (provider orders and nursing flowsheet documentation) in an electronic health record. Orders and nursing documentation were simultaneously present for only 42.3% of patient rooms with contact precaution signs, and 17.8% of rooms with signs had neither orders nor nursing documentation. PMID- 21623182 TI - The utility of ERG/P63 double immunohistochemical staining in the diagnosis of limited cancer in prostate needle biopsies. AB - Diagnosis of limited cancer can be challenging in prostate needle biopsies, and immunohistochemistry is commonly used in such settings. Recently, TMPRSS2:ERG gene rearrangement was found to be highly specific for and detected in approximately 50% of prostate cancer. Positive immunohistochemical staining with a novel anti-ERG antibody highly correlated with TMPRSS2:ERG gene rearrangement status. We developed a double immunohistochemical staining containing both erythroblastosis virus E26 oncogen (ERG) and basal cell marker P63 antibodies and evaluated its use in the diagnosis of limited cancer in prostate needle biopsies. A total of 77 prostate needle biopsies containing cancer occupying <1 mm of the length of only 1 core of the entire biopsy set were stained with the double stain containing ERG and P63 antibodies. ERG positivity and its staining intensity in cancerous and other noncancerous lesions were evaluated. ERG expression was detected in 42% (32 of 77) of cases, with strong, moderate, and weak staining intensity in 72%, 16%, and 12% of cases. The staining was uniform in 84% of cases and heterogeneous in 16% of cases with different staining intensities in >10% of cancerous cells. High-grade prostatic intraepithelial neoplasia was present in 17 cases, and in 5 (29%) cases ERG was positive in high-grade prostatic intraepithelial neoplasia glands, which were all immediately adjacent to or intermingled with ERG-positive cancerous glands. In 4 additional cases, positive ERG staining was found in morphologically benign glands, which were also immediately adjacent to or intermingled with ERG-positive cancerous glands. All other benign lesions distant from cancerous glands, including simple and partial atrophy, were negative for ERG. P63 was negative in all cancerous glands and positive in noncancerous lesions. The P63/ERG double immunostain combines the high sensitivity of P63 and the high specificity of ERG and may be potentially useful in the work-up of difficult prostate biopsies. The high specificity of ERG for the presence of cancer may have important implications for prostate biopsy interpretation and needs to be further validated in larger prospective studies. PMID- 21623183 TI - Primary human tumor xenografted models ('tumorgrafts') for good management of patients with cancer. AB - The treatment of cancer is continually improving as a result of a better understanding of oncogenesis and the development of new targeted compounds. Early clinical trials evaluating such candidate compounds require a large number of patients, and are expensive, time consuming, and expose patients to certain risks. To select the most effective molecules, preclinical investigation of antitumor compounds is an important step in the drug development process. Three main categories of preclinical cancer models are generally used in preclinical investigations, namely genetically engineered models, xenografts derived from human tumor cell lines, and human tumor fragments from patients implanted directly into immunodeficient mice, known as tumorgrafts. The establishment of tumorgrafts constitutes a long-term process consisting of various steps, in which the final objective is to show that the validated model accurately reproduces human cancer, with a high predictive value of therapeutic efficacy (regardless of the type of treatment), and closely mimics clinical situations frequently observed in patients with cancer, such as resistance to standard treatments, metastases, and relapse after initial therapies (involving residual tumor initiating cells). The aim of this study is therefore to discuss the proposed criteria for the establishment and validation of preclinical models of human cancers that should be used for further pharmacological assessments. PMID- 21623184 TI - Relationship of cell bearing EBER and p24 antigens in biopsy-proven lymphocytic interstitial pneumonia in HIV-1 subtype E infected children. AB - Lymphocytic interstitial pneumonia (LIP) is an uncommon histopathologic entity characterized by infiltration of the interstitium and alveolar spaces of the lung by lymphocytes and other lymphoid elements. An increased incidence of LIP has been seen in the pediatric population, especially in children with acquired immune deficiency syndrome. Our previous study supports the notion that Langerhans cells (LCs) are reservoirs for Epstein-Barr virus (EBV) in lungs of human immunodeficiency virus (HIV) subtype E-infected pediatric LIP. To further understand the pathogenesis of LIP, we studied the relationship between EBV, the suggested causative agent of LIP and HIV-1 capsid protein p24, which play an important role in the interaction with host proteins during HIV-1 adsorption, membrane fusion, and entry in surgical lung biopsy-proven LIP from 9 vertically HIV subtype E-infected pediatric patients. The dominant microscopic feature of LIP demonstrated widespread widening of alveolar septum by mononuclear inflammatory cell infiltrate, mainly composed of mature lymphocytes and plasma cells surrounding airways and expanding to the lung interstitium. EBV-encoded RNA (EBER) in situ hybridization (ISH) and p24 immunohistochemistry, performed on formalin-fixed, paraffin-embedded tissue from open lung biopsy specimens, revealed positive intranuclear EBER signals and intracytoplasmic immunostains for p24 core protein in all 9 LIP cases. By combining ISH and immunohistochemistry, these results suggest that (i) EBV/p24-carrying cells are likely involved in the development of LIP, either directly or indirectly; (ii) LCs and related dendritic cells are the main reservoir of both EBV and HIV subtype E in pediatric LIP and possibly LCs may play an important role in the recruitment of inflammatory cell infiltrates, especially T cells into these tissues; (iii) coexpression of EBV/p24 in bronchioalveolar epithelium supports the hypothesis that these cells serve as a reactivation source for both viruses to achieve greater quantities in alveolar septum and interstitium around bronchioles. These results indicate a strong association between the presence of HIV core protein p24 and expression of EBV RNA transcripts (EBER). Interactions between LCs and related dendritic cells together with T cells are important for effective HIV and EBV replications. The coexpression of both viruses could be related to the evolution of pediatric LIP in HIV subtype E infection. PMID- 21623185 TI - Immunohistochemical determination of ETS-1 oncoprotein expression in urothelial carcinomas of the urinary bladder. AB - ETS-1 protooncogene is an important transcription factor that plays a role in the regulation of physiological processes, such as cell proliferation and differentiation. ETS-1 is thought to be related to the growth of carcinoma cells by its regulation of the transcription of matrix metalloproteinases and urokinase type plasminogen activator. In this study, we aimed to investigate the expression pattern of ETS-1 oncoprotein in urothelial carcinomas of the urinary bladder and determine its relationship with histopathologic parameters, including tumor grade and stage. One hundred six specimens of urothelial carcinoma and a total of 14 normal urothelium were analyzed immunohistochemically with anti-ETS-1 monoclonal antibody. The normal urothelium showed positive ETS-1 immunostaining. ETS-1 expression remained high in low-grade and noninvasive tumors, whereas it frequently decreased in high-grade or invasive carcinomas. Interestingly, ETS-1 was highly expressed in the basal cell layer of the noninvasive urothelial carcinomas. ETS-1 expression showed a strong negative correlation with the tumor grade (P<0.001; r, -0.67) and stage (P<0.001; r, -0.75). The nonmuscle-invasive tumors (pTa+pT1) and noninvasive tumors (pTa) had significantly higher ETS-1 expression than the muscle-invasive tumors (pT2; P<0.001) and invasive tumors (pT1+pT2; P<0.001), respectively. Results of our study show that decreased ETS-1 expression is significantly associated with high grade and advanced stage in urothelial carcinomas of the urinary bladder, and that the downregulation of ETS 1 expression may be a marker of the aggressiveness of such malignancies. PMID- 21623186 TI - Using mean duration and variation of procedure times to plan a list of surgical operations to fit into the scheduled list time. AB - BACKGROUND AND OBJECTIVE: It is important that a surgical list is planned to utilise as much of the scheduled time as possible while not over-running, because this can lead to cancellation of operations. We wished to assess whether, theoretically, the known duration of individual operations could be used quantitatively to predict the likely duration of the operating list. METHODS: In a university hospital setting, we first assessed the extent to which the current ad-hoc method of operating list planning was able to match the scheduled operating list times for 153 consecutive historical lists. Using receiver operating curve analysis, we assessed the ability of an alternative method to predict operating list duration for the same operating lists. This method uses a simple formula: the sum of individual operation times and a pooled standard deviation of these times. We used the operating list duration estimated from this formula to generate a probability that the operating list would finish within its scheduled time. Finally, we applied the simple formula prospectively to 150 operating lists, 'shadowing' the current ad-hoc method, to confirm the predictive ability of the formula. RESULTS: The ad-hoc method was very poor at planning: 50% of historical operating lists were under-booked and 37% over-booked. In contrast, the simple formula predicted the correct outcome (under-run or over-run) for 76% of these operating lists. The calculated probability that a planned series of operations will over-run or under-run was found useful in developing an algorithm to adjust the planned cases optimally. In the prospective series, 65% of operating lists were over-booked and 10% were under-booked. The formula predicted the correct outcome for 84% of operating lists. CONCLUSION: A simple quantitative method of estimating operating list duration for a series of operations leads to an algorithm (readily created on an Excel spreadsheet, http://links.lww.com/EJA/A19) that can potentially improve operating list planning. PMID- 21623187 TI - The oral cancer knowledge of dentists in Northern Germany after educational intervention. AB - The objective of this study was to assess the knowledge about oral cancer, in particular about diagnostic items and risk factors, after educational intervention by means of a standardized survey. On the basis of the evaluation at baseline, an educational programme consisting of different approaches (for instance, oral presentations, poster, DVD and a brochure) was designed. In May 2009, the questionnaire 'Practices and opinions about oral cancer' focusing on the knowledge about diagnostic factors and risk factors for oral cancer was mailed to every dentist in Schleswig-Holstein, Germany (n = 2280). The results were based on 394 of 2280 questionnaires. After a 1-year educational intervention, the dentists' overall knowledge about diagnostic procedures and risk factors for oral cancer had improved. The low baseline results for diagnostic procedures and risk factors showed the highest improvement rate immediately after the intervention. Within the reevaluation subgroups, responders who had attended a continuing education course during the intervention period showed better results compared with the whole sample of the reevaluation. The results showed that a structured educational programme with different approaches increases dentists' knowledge about diagnostic procedures and risk factors. This intervention may not only give useful recommendations for further educational courses, but emphasizes the necessity of attending continuous further educational courses. PMID- 21623188 TI - Exhaled nitric oxide in patients with early glottic cancer. AB - Nitric oxide (NO) may contribute to the mutagenic processes and has been extensively studied as a noninvasive marker of neoplastic processes occurring in the airways. It has been suggested that an increased expression of inducible NO synthase might appear as an important molecular event involved in the early stage of laryngeal carcinogenesis. The aim of this study was to determine NO levels in the exhaled air of patients with early stage glottic carcinoma. Exhaled bronchial NO was measured using a Niox Mino analyzer in 28 patients with squamous cell glottic carcinoma (T1-T2N0M0) and in 30 healthy controls. There were no significant differences in exhaled NO level between the healthy controls and T1 T2N0M0 patients with glottic cancer or between the subgroups (T1 vs. T2). It seems that early glottic cancer is not associated with significant changes in the exhaled NO level. Early diagnosis of cancer is desirable, however this study does not prove that exhaled NO may be a useful biomarker in patients with early stage squamous cell glottic carcinoma. PMID- 21623189 TI - Gambles of gastroenterology. AB - BACKGROUND: Diagnostic and therapeutic procedures of gastroenterology can frequently tax a patient's health. In addition to the risk of side-effects such procedures may also reduce the patient's well-being. The probability of achieving cure depends on the interplay between the patient's initial health status and the expected efficacy of the gastroenterological interventions. METHODS: The mathematical concept of the 'gambler's ruin' from probability theory is used to model the interplay among the various factors affecting clinical outcome. RESULTS: The model illustrates in a quantitative manner how the physician's ultimate success in achieving cure depends on a given patient's initial health status, and the characteristics of the interventional procedures used. The analysis also illustrates that in some dire clinical situations, risking the patient's life in one single and bold step can yield a better overall outcome than trying to advance the patient's health by multiple small and cautious steps. CONCLUSION: Using the gambler's ruin as a model for medical risk taking could contribute to medical teaching and promote the understanding of the some general principles that shape many clinical decisions in gastroenterology. PMID- 21623190 TI - The aetiology of symptomatic gallstones quantification of the effects of obesity, alcohol and serum lipids on risk. Epidemiological and biomarker data from a UK prospective cohort study (EPIC-Norfolk). AB - OBJECTIVE: The development of gallstones is influenced by obesity and alcohol. This study aimed to precisely quantify these risks and investigate whether the aetiological mechanism may involve serum lipids, for the first time using a European prospective cohort study. METHODS: The European Prospective Investigation into Cancer-Norfolk, recruited 25 639 men and women, aged 40 to 74 years, between 1993 and 1997. At enrolment weight, height and alcohol intake were recorded and nonfasting blood samples taken to measure serum triglycerides, cholesterol, high-density lipoproteins and low-density lipoproteins. The cohort was monitored for 14 years for symptomatic gallstones. Cox regression estimated sex-specific hazard ratios (HRs) for symptomatic gallstones adjusted for covariates. RESULTS: Symptomatic gallstones developed in 296 people (67.9% women). For each additional unit of BMI, the HR in men was 1.08 [95% confidence interval (CI)=1.02-1.14]; in women the HR was 1.08 (95% CI=1.06-1.11). Every unit of alcohol consumed per week decreased risk in men by 3% (HR=0.97, 95% CI=0.95 0.99) with no effect in women. Serum triglycerides increased risk in men (highest vs. lowest quarter HR=2.02, 95% CI=1.03-3.98) and women (HR=2.43, 95% CI=1.52 3.90). Increased high-density lipoprotein was associated with a decreased risk in men (highest vs. lowest quarter HR=0.22, 95% CI=0.09-0.52) and women (HR=0.55, 95% CI=0.36-0.85). No effects were found for serum cholesterol and low-density lipoprotein. CONCLUSION: Obesity and alcohol influence gallstone formation, possibly in part through their effects on serum lipids. Reducing obesity may prevent gallstones in the population, as 38% of incident cases of gallstones were associated with a BMI of more than 25. PMID- 21623191 TI - Usefulness of a novel serum proteome-derived index FI-PRO (fibrosis-protein) in the prediction of fibrosis in chronic hepatitis C. AB - BACKGROUND: Liver biopsy is an imperfect standard for the assessment of chronic hepatitis C liver fibrosis. In this study, the diagnostic role of proteome derived protein markers and the usefulness of a protein-based index were assessed. METHODS: Characteristics, clinical biochemistry, and protein markers of patients with chronic hepatitis C from a study (n=62) and validation group (n=73) were statistically assessed according to fibrosis severity. Multivariate models were built using linear discriminant analysis for the prediction of minor fibrosis (F0-F1), moderate fibrosis (F2-F3), and cirrhosis (F4). The best model was validated and diagnostic performance was compared with the aspartate aminotransferase-to-platelet ratio index based on their receiver operator characteristic curves. RESULTS: Statistical analysis resulted in significant outcomes for both clinical and protein markers. The best multivariate model was based on four protein markers: alpha-2-macroglobulin (A2M), haptoglobin, hemopexin, and galectin-3-binding protein. A2M and hemopexin were the primary predictors according to this model. A novel index A2M/hemopexin [fibrosis-protein (FI-PRO) index] showed a diagnostic performance rate of 0.80-0.92 for the detection of significant fibrosis (F2-F4) and advanced fibrosis (F3-F4) in the validation group, which was better compared with aspartate aminotransferase-to platelet ratio index. FI-PRO had an overall positive predictive value of 86% for significant fibrosis and a negative predictive value of at least 90% for advanced fibrosis. CONCLUSION: Proteome-derived protein markers were successfully implemented in clinical diagnosis of hepatitis C fibrosis, which resulted in the FI-PRO index. The efficiency and usability of FI-PRO should be validated in large scale, prospective studies. PMID- 21623193 TI - The case reported as bilateral Sertoli-Leydig cell tumors in a 61-year-old woman with uterine aplasia may instead represent complete androgen insensitivity syndrome. PMID- 21623194 TI - Dedifferentiated chondrosarcoma arising in a mature cystic teratoma of the ovary: a case report and review of the literature. AB - Malignant transformation of a mature cystic teratoma of the ovary is rare, occurring in approximately 2% of all cases. The most common malignancy arising in mature cystic teratoma is squamous cell carcinoma. Much less frequently, the malignant transformation is represented by sarcomas. Dedifferentiated chondrosarcoma usually develops in bone. There has been no case of a dedifferentiated chondrosarcoma arising in mature cystic teratoma of the ovary since the establishment of this diagnostic entity. This is a report of a definitive dedifferentiated chondrosarcoma arising in a mature cystic teratoma of the ovary, presenting clinicopathologic features. PMID- 21623195 TI - Stem cells in human endometrium and endometrial carcinoma. AB - This study reviews the current knowledge about endometrial somatic stem cells and endometrial cancer stem cells. It describes the main features of somatic stem cells, such as high proliferative potential, self renewal, differentiation into 1 or more lineages, retention of a DNA synthesis label (BrdU), and some methods to identify them (Hoechst dye exclusion test, immunophenotyping). The most likely markers for endometrial somatic stem cells (Oct-4, Musashi-1, CD31, CD34, and CD144) are also mentioned. The study also reviews the literature regarding endometrial cancer stem cells. Results obtained by evaluations of the side population in endometrial cancer cell lines and studies on putative cancer stem cell markers are also discussed. The possible roles of endometrial cancer stem cells in metastasis and resistance to anticancer treatment are also mentioned. PMID- 21623196 TI - Clinicopathologic correlations between human papillomavirus 16 infection and Beclin 1 expression in human cervical cancer. AB - Our earlier study showed that the autophagy gene Beclin 1 could affect cell proliferation in a cervical cancer HeLa cell line. In this study, we examined Beclin 1 protein expression in 81 specimens of cervical squamous carcinoma by immunohistochemistry. Meanwhile, we detected E6 and E7 genes of human papillomavirus 16 in these tissues by polymerase chain reaction. Beclin 1 expression significantly decreased in samples of malignant cervical cancer tissues than in those of normal or cervical intraepithelial neoplasia tissues. The expression of Beclin 1 was associated with pelvic lymph node metastasis and histological grade, but did not correlate with International Federation of Gynecology and Obstetrics stage, age, depth of cervical infiltration, tumor size, and gross type of cervical lesion. The expression of Beclin 1 was not obviously correlated with E6 and E7 genes statistically. Therefore, decreased expression of Beclin 1 may be related to tumorigenesis and the development of cervical cancer, but is not significantly relevant with human papillomavirus 16 infection. PMID- 21623197 TI - Mucinous cystadenoma coexisting with adult granulosa cell tumor in the ovary: is it a composite tumor or heterologous mucinous elements in a granulosa cell tumor? AB - We herein report a rare tumor combination of mucinous cystadenoma and an adult granulosa cell tumor in the same ovary of a 50-year-old woman. Ultrasound examination showed a multicystic mass in the left ovary. Histologically it showed 2 components that were intimately admixed. One was composed of cysts lined by mucinous epithelium of the intestinal type that was strongly positive for cytokeratin-20 and cytokeratin-7. The other was a granulosa cell tumor of the adult type in which the tumor cells showed microfollicular, sheets, and trabecular patterns. They were positive for calretinin, alpha-inhibin, and CD99. There are many theories for the histogenesis of such a combination. After discussing all the possibilities we conclude that the mucinous component is probably a heterologous mucinous differentiation within an adult granulosa cell tumor because of the intimate admixture of the 2 components and the mucinous epithelium being of the intestinal type. PMID- 21623198 TI - Borderline mucinous endocervical tumor as a link between the endometriotic cyst and ovarian primary squamous cell carcinoma. PMID- 21623199 TI - Screening for NLRP7 mutations in familial and sporadic recurrent hydatidiform moles: report of 2 Tunisian families. AB - A familial or sporadic recurrent hydatidiform mole is a rare autosomal recessive condition that has been associated with biallelic mutations in the nucleotide binding, leucine-rich repeat, pyrin domain 7 (NLRP7) gene (19q13.42). Cases from different ethnic origins have been reported earlier. Here we report the first Tunisian patients: 2 sisters with homozygous NLRP7 mutations (p.E570X) and 1 sporadic case with no mutation in NLRP7. Our results extend the number of familial recurrent reproductive wastages due to mutations in NLRP7. We suggest that mutations screening of NLRP7 could be proposed more systematically in women with recurrent pathologic pregnancy outcomes of unknown origin. The rare cases with a typical clinical picture, which were not related to NLRP7 mutation as in our sporadic case, should be investigated more to identify the causative gene. PMID- 21623200 TI - A quality process study of lymph node evaluation in endometrial cancer. AB - Our objective was to analyze the reported lymph node counts between surgeons, histology prosectors, and pathologists using a cohort of patients enrolled on a national protocol that standardized surgical intent.This is a retrospective review of patients with uterine cancer who underwent a standardized formal staging procedure as dictated by a National Cancer Institute sponsored protocol. Patients were staged using the International Federation of Gynecology and Obstetrics 1988 guidelines. All patients required a hysterectomy, bilateral salpingo-oophorectomy, and bilateral pelvic and para-aortic lymphadenectomy. Lymphadenectomy specimens were separated by the following regions: external iliac, obturator, common iliac, and periaortic. Lymph node counts were analyzed by region, surgeon, histology prosector, and pathologist.There were 78 patients enrolled in the protocol during the study period. Of them, 72 (92%) patients met the inclusion criteria. A total of 2397 lymph nodes were counted, with an average total number of 33 (SD=9) lymph nodes dissected per patient. Surgeons A, B, and C had an average lymph node count of 32, 33, and 35, respectively, with no significant difference in mean node count (P=0.66). Prosectors 1 to 4 dissected an average of 34, 33, 28, and 35 lymph nodes, respectively (P=0.091). There were 2 pathologists with >= 10 cases. Their mean lymph node counts were 35 and 30, respectively, with no significant difference in mean node count (P=0.079).This systematic review did not identify a discrepancy in nodal count among surgeons, prosectors, or pathologists at our institution. The methods used may be helpful in structuring interdepartmental reviews for completeness of nodal dissections in cases where surgical intent has been standardized. PMID- 21623201 TI - Biomarker expression in pelvic high-grade serous carcinoma: comparison of ovarian and omental sites. AB - Neoadjuvant therapy has an emerging role in the treatment of high-stage ovarian carcinoma. Some ovarian carcinoma subtypes do not respond well to standard chemotherapy, making accurate subtype diagnosis before starting therapy important. This diagnosis is frequently based on omental biopsy specimens. In particular, with very small biopsies, immunostaining for diagnostic biomarkers may be needed. To assess intratumoral heterogeneity of biomarker expression in pelvic high-grade serous carcinoma, we compared the expression of a set of 10 biomarkers between ovarian and omental sites. Tissue microarrays were constructed from 123 high-grade serous carcinomas with paired ovarian and omental tumor samples. These samples were stained with biomarkers that have been used in ovarian carcinoma subtype diagnosis (WT1, TP53/p53, MUC16/CA125, CDKN2A/p16), and with biomarkers of the tumor microenvironment (CD8, CD163, SPARC, PDGFRB), cell adhesion (CDH1/E-Cadherin), and proliferation (Ki67) as well. Expression frequencies in samples from the 2 sites were compared, as was concordance at the 2 sites for individual tumors. The 2 markers of desmoplastic stromal response (PDGFRB, SPARC) were more frequently expressed in the omentum compared with the ovary (P<0.001; McNemar test). The other 8 markers did not show a significant difference in the frequency of expression between sites. Within individual cases, some markers such as Ki67 and CDKN2A showed variability, indicating that these markers are affected by intratumoral heterogeneity. The intratumoral variability for MUC16, TP53, and WT1 was modest. Commonly used diagnostic markers, such as TP53 and WT1, show little variability between ovarian and omental sites, suggesting that they can be successfully used in small biopsy specimens from extraovarian sites. In contrast, markers of host stromal response do vary between sites, suggesting a biologic difference of the microenvironment at different sites that should be taken into account when tissue-based research is carried out. PMID- 21623202 TI - Assessing the impact of polysomy-17 on HER2 status and the correlations of HER2 status with prognostic variables (ER, PR, p53, Ki-67) in epithelial ovarian cancer: a tissue microarray study using immunohistochemistry and fluorescent in situ hybridization. AB - Although HER2 overexpression and Her2 amplification have been noted in breast and a variety of human cancers, we report here for the first time the impact of polysomy-17 on HER2 status and the correlations between HER2 status and other prognostic factors in patients with epithelial ovarian cancers (EOC).We analyzed HER2, estrogen receptor (ER), progesterone receptor (PR), p53, and Ki-67 protein overexpressions by immunohistochemistry (IHC) and determined Her2 gene amplification by fluorescence in situ hybridization (FISH) in 27 tissue microarray samples from EOC patients.We achieved 100% positive concordance (3/3) and 100% negative concordance (19/19) between HER2 testing by IHC and FISH. Both the total Her2 gene copies and FISH scores increased significantly in a stepwise order through the negative, equivocal, and positive HER2 IHC result categories in all 27 cases (P=0.001, P=0.001), and still increased significantly in 18 nonpolysomy-17 cases (P=0.007 and 0.013) after the exclusion of 9 polysomy-17 cases. HER2 protein expression is inversely correlated with both ER (P=0.002) and PR expressions (P=0.046). Her2 gene amplification is inversely correlated with ER expression (P=0.007) but not with PR expression (P=0.106).This study showed extremely high positive and negative concordances between Her2 FISH and HER2 IHC assays. Polysomy-17 is insufficient for causing a significant impact on the relationship between HER2 testing by IHC and FISH in EOC. ER and PR expressions were inversely correlated with HER2 protein expression. In addition, ER but not PR expression is inversely correlated with Her2 gene amplification. PMID- 21623203 TI - Mullerian adenosarcomas with unusual growth patterns: staging issues. AB - Uterine adenosarcomas are uncommon mixed Mullerian neoplasms, most commonly arising in the uterine corpus. A new Federation of International Gynecologic Organization staging system for these tumors has recently been implemented. This staging system is an improvement on the earlier generic application of the 1988 Federation of International Gynecologic Organization staging system for endometrial cancer to adenosarcoma. Herein, we report 3 uterine adenosarcomas with unusual features. For 2 of these, no specific staging guidelines are provided by either the earlier or, more importantly, the new staging system. The first case is of an adenosarcoma arising in the eutopic endometrium with involvement of underlying adenomyosis without myometrial invasion; the second originated in a mural adenomyoma in the absence of eutopic endometrial involvement; and the third case encompassed synchronous endometrial and extrauterine (peritoneal) neoplasms. Such cases are rare, and there is insufficient evidence to be definitive about staging. Thus, we suggest a descriptive reporting strategy for adenosarcomas with these unusual features. We also propose a reporting nomenclature for such cases to ensure standardization such that they can be adequately recorded in synoptic reporting protocols. This will facilitate reliable data collection such that an evidence-based staging system for these scenarios may be derived. PMID- 21623204 TI - Liesegang rings in an ovarian endometriotic cyst fluid aspirate. PMID- 21623205 TI - Usefulness of p16ink4a, ProEX C, and Ki-67 for the diagnosis of glandular dysplasia and adenocarcinoma of the cervix uteri. AB - Although the diagnostic criteria of in-situ and invasive adenocarcinomas of the cervix uteri are well established, the differentiation from benign mimics may be difficult and the morphologic features of the precursors of endocervical adenocarcinoma are still debated. In this study, we evaluated the usefulness of p16ink4a (p16), ProEX C, and Ki-67 for the diagnosis of endocervical adenocarcinoma and its precursors. Immunohistochemistry with p16, ProEX C, and Ki 67 was performed in 82 glandular lesions including 15 invasive adenocarcinomas, 29 adenocarcinomas in situ (AIS), 22 non-neoplastic samples, and 16 cases of glandular dysplasia (GD), which showed significant nuclear abnormalities but did not meet the diagnostic criteria for AIS. The immunohistochemical expression pattern was scored according to the percentage of the stained cells (0, 1+, 2+, and 3+ when 0% to 5%, 6% to 25%, 26% to 50%, and more than 50% of the cells were stained, respectively) and was evaluated for each antibody. p16 was at least focally expressed (1+ or more) in 14 of 15 invasive adenocarcinomas, in all AIS and in 7 negative samples. ProEX C and Ki-67 both scored 1+ or more in all adenocarcinomas and AIS and in 8 and 6 negative samples, respectively. Of the GD 15, 14, and 15 expressed p16, ProEX C, and Ki-67, respectively. The score differences between neoplastic and non-neoplastic samples were highly significant for each marker (P<0.001); however, the score distribution by marker differed significantly only in GD (P=0.006) in which, compared with the other markers, p16 showed more often a 3+ pattern. Our study shows that p16, Ki-67, and ProEX C may be helpful for the diagnosis of glandular lesions of the cervix uteri and may also improve the diagnostic accuracy of endocervical GD. In particularly problematic cases, the combination of p16 and a proliferation marker can provide additional help for the interpretation of these lesions. PMID- 21623206 TI - Borderline ovarian mucinous neoplasm recurring as small cell carcinoma of hypercalcemic type: evidence for an epithelial histogenesis and relationship with ovarian mucinous tumors for this enigmatic neoplasm. AB - Ovarian small cell carcinoma of hypercalcemic type (OSCCHT) is an uncommon neoplasm of uncertain histogenesis. As far as we are aware, this neoplasm has never been reported in association with another primary ovarian tumor. We report a case in a 33-year-old patient in whom an extraovarian pericolonic neoplasm with the morphological features and immunohistochemical profile of OSCCHT developed 5 years after removal of an ovarian mucinous borderline tumor of the intestinal type. Together with the observation that mucinous epithelium is seen in some OSCCHTs, this case raises the possibility that this enigmatic neoplasm is of epithelial origin and is related to primary ovarian mucinous neoplasms. The pericolonic tumor exhibited an unusual immunophenotype with positive staining with CD99 and FLI-1, suggesting the possibility of a neoplasm in the Ewing family of tumors. This was excluded by molecular studies that showed no evidence of EWS/FLI-1 or EWS/ERG translocation. PMID- 21623207 TI - Comparative analysis of the ERalpha/ERbeta ratio and neurotensin and its high affinity receptor in myometrium, uterine leiomyoma, atypical leiomyoma, and leiomyosarcoma. AB - Deregulated steroids are involved in different hormone-dependent tumors, including benign and malignant uterine neoplasms. Leiomyomas (LM) are estrogen and progesterone-dependent benign tumors, whereas "bizarre or atypical LMs" (AL) are considered a subgroup of LM and clinically benign, although their malignant potential is suspect. Uterine leiomyosarcomas (LMS) are malignant smooth muscle tumors, and ovarian steroids may control their growth. Estrogen effects are mediated by 2 receptors, estrogen receptors (ER) alpha and beta, and the ratio of both receptors seems to be a critical parameter in the estrogen-mediated carcinogenic process. Estradiol induces the expression of neurotensin (NTS), and the coupling of this peptide with its high-affinity receptor, NTS1, has been involved in the regulation of tumoral cell growth. Given the importance of these markers in tumor development, we aim to determine the status of ERalpha and ERbeta in the myometrium and LM, AL, and LMS, concomitantly with the expression of NTS/NTS receptor 1 in these tumors. For that purpose, we use immunohistochemistry for all markers analyzed and in-situ hybridization to detect NTS mRNA. These data suggest that LMS are estrogen-dependent tumors, which may use NTS as an autocrine growth factor. In addition, the phenotype of AL with regard to ERalpha and ERbeta status and NTS expression is closer to LMS than LM; thus, a potential malignization of this tumor is feasible. PMID- 21623208 TI - Intraplacental choriocarcinoma without associated maternal or fetal metastases. PMID- 21623209 TI - Comparison of FIGO 1989 and 2009 recommendations on staging of endometrial carcinoma: pathologic analysis and cervical status in 123 consecutive cases. AB - Incidences of endometrial carcinoma (EC) among women of Western countries is increasing, reaching a level of 18/100,000. In 2009, the International Federation of Gynecology and Obstetrics (FIGO) proposed a new staging system in EC. The purpose of this study included the evaluation of distribution of EC in categories of age, histologic grade, and surgical staging according to the 1989 and the 2009 FIGO guidelines. The original staging assessments have been updated to reflect the current staging system in 123 consecutive patients. Statistical analysis was carried out. The median age of patients was 61 years. A comparison of old and new staging systems shows that a significant number of patients moved to stage I: 78.05%, versus 56.91% based on the 1989 classification (P=0.044). The number of patients in stage II changed as well: 9.76% according to new staging system and 30.89% by definitions of FIGO 1989 (P=0.001). Of patients in stage II, 21.1% had G1 tumors according to the old versus 8.3% by the new classification (P=0.001). We have not identified any associations between the histologic status of a cervix and EC. The most common type of EC is the endometrioid subtype, found in 87.8% of patients. We have noted a significant association between the tumor grade and cervical stromal infiltration. The new classification system for EC seems to be an improved staging instrument. Having up to 80% of patients with endometrial cancer in stage I and following them, might elucidate the impact of current staging on survival and life quality. PMID- 21623210 TI - Renal effects of bicarbonate versus saline infusion for iso- and lowosmolar contrast media in rats. AB - OBJECTIVE: Hydration is widely accepted as an effective measure to prevent contrast media (CM)-induced acute kidney injury (AKI). Whether bicarbonate (NaHCO3) infusion has a greater effect than saline is disputed. Effective prevention of CM-induced AKI by NaHCO3 has been found by several clinical trials. However, others found either no effect or an enhanced incidence of CM-induced AKI after giving NaHCO3. Because of their different tubular resorption, NaHCO3 and saline may have a different capacity to flush the nephron. In this study, we compare the magnitudes by which NaHCO3 and saline can enhance urinary flow, prevent a decline in glomerular filtration rate (GFR), and limit an increase in urine viscosity, as caused by CM administration. MATERIALS AND METHODS: Prewarmed (37 degrees C) CM were administered as 1.5 mL bolus into the thoracic aorta of anesthetized rats. Following 2 CM were studied: iso-osmolar iodixanol (320 mg I/mL) and low-osmolar iopromide (370 mg I/mL). Four protocols (n = 7 rats per protocol) were followed: (1) saline + iodixanol, (2) saline + iopromide, (3) NaHCO3 + iodixanol, and (4) NaHCO3 + iopromide. Isotonic saline or NaHCO3 were infused at a rate of 4 mL/h per kg BM, initiated 60 minutes before CM administration and continued throughout the observation period of 100 minutes. Urine volume was measured gravimetrically, urine viscosity was measured by a microviscometer, and GFR was determined by creatinine clearance. RESULTS: As compared with saline infusions, NaHCO3 infusions did not significantly alter the effects that iodixanol and iopromide exerted on urine flow rate, urine viscosity, and GFR. In the iopromide protocols, CM-induced increase in urine flow was about 50% greater than in the respective iodixanol protocols. Conversely, in the iodixanol protocols, urine viscosity was up to 10-fold greater than in the respective iopromide protocol. In the iodixanol protocols, GFR decreased transiently (10-30 min post-CM) by up to 50%, whereas GFR did not decrease in the iopromide protocols. CONCLUSION: Infusing either saline or NaHCO3 seems to make little difference with regard to urine flow, urine viscosity, and GFR. However, the CM used has a significant effect on these measures. Iopromide enhances urine flow by a greater magnitude than iodixanol, whereas the latter increases urine viscosity to a larger degree than iopromide and transiently decreases GFR. PMID- 21623211 TI - Safety of gadobutrol, a new generation of contrast agents: experience from clinical trials and postmarketing surveillance. AB - OBJECTIVE: To assess the clinical safety and tolerability of the macrocyclic contrast agent gadobutrol (Gadovist/Gadavist) overall and in specific patient populations based on clinical trials and postmarketing experience. MATERIALS AND METHODS: In total, 5545 patients enrolled in 34 prospective clinical studies were evaluated in an integrated analysis of safety. Of all enrolled patients, 4549 received gadobutrol at a dose of <= 0.09 mmol/kg body weight to a maximum of 0.51 mmol/kg body weight, with most patients (53.5%) receiving the recommended dose of >0.09 to 0.11 mmol/kg body weight. Data include comparisons with other extracellular contrast agents and subgroup analyses in pediatric patients, and patients with allergic disposition, renal impairment, hepatic impairment, or cardiovascular disease. Furthermore, worldwide postmarketing safety surveillance results, including nephrogenic systemic fibrosis reports, based on more than 5.7 million estimated applications are described. RESULTS: One or more adverse events (AEs) assessed as related to the administration of gadobutrol were reported by 182 (4.0%) of the 4549 patients who participated in clinical trials. This is comparable to the incidence observed with the comparator contrast agents (74/1844 patients, 4.0%). The most common AEs, independent of drug relationship, were headache, nausea, feeling hot, and dysgeusia. The favorable safety profile of gadobutrol was also demonstrated in the following specific subpopulations in whom similar incidence rates were seen: pediatric patients aged 2 to 17 years (8/138 patients, 5.8%), patients with severe or moderate renal impairment (9/366 patients, 2.5%), patients with severe or moderate hepatic impairment (9/214 patients, 4.2%), and patients with cardiovascular disorders (42/1506 patients, 2.8%). Having been established in controlled clinical trials, this safety profile was also confirmed by postmarketing surveillance data. With more than 5.7 million estimated administrations of gadobutrol, a total of 1175 (0.02%) suspected adverse drug reactions have been reported. The most serious adverse reactions seen in postmarketing surveillance included rare reports of cardiac arrest, respiratory arrest, anaphylactoid shock, and nephrogenic systemic fibrosis. Incidence and type of AEs from postmarketing surveillance were consistent with the established safety profile. CONCLUSION: The comprehensive analysis of safety data obtained from 34 clinical studies demonstrates that gadobutrol has an excellent safety profile and a positive benefit risk profile when used in patients in need of contrast-enhanced magnetic resonance imaging. Gadobutrol was well tolerated by adults, by children, by patients with impaired liver or kidney function, and by patients with cardiovascular disease. The favorable safety profile is confirmed by the available postmarketing surveillance data and is compared with that of other gadolinium-based contrast agents. PMID- 21623212 TI - Pharmacokinetics and imaging properties of Gd-EOB-DTPA in patients with hepatic and renal impairment. AB - OBJECTIVES: : The purpose of this study was to determine the pharmacokinetics (PKs), imaging properties, and safety of the liver-specific magnetic resonance (MR) imaging contrast agent gadoxetic acid disodium (Gd-EOB-DTPA) in subjects with various levels of hepatic impairment, renal impairment, or coexisting hepatic and renal impairment. MATERIALS AND METHODS: : In this single-center, open-label, parallel-group study, patients with varying degrees of renal and/or hepatic impairment were compared with healthy subjects matched for age, gender, and weight (control group). All subjects received a single intravenous bolus of Gd-EOB-DTPA (Primovist, Eovist, EOB-Primovist) 25 MUmol/kg body weight. Samples of serum, urine, and feces were collected for PK analysis. MR imaging was performed before dosing and at preset times after dose administration to determine enhancement relative to predose signal intensity values. Safety was assessed by monitoring adverse events, laboratory values, vital signs, cardiac rhythm, oxygen saturation, and by physical examination findings. RESULTS: : Gd EOB-DTPA was well tolerated by all subjects. Total clearance of Gd-EOB-DTPA did not significantly change in patients with mild and moderate hepatic impairment (Child-Pugh A and B), compared with the control group. Mean urinary excretion was increased and mean fecal excretion was decreased in patients with hepatic impairment. Renal excretion was increased to between 72% and 96% of the dose administered in patients with very high bilirubin levels (>3 mg/dL), compared with 48% in the control group. Total clearance of Gd-EOB-DTPA was significantly reduced to 140 +/- 45 mL/min and terminal elimination half-life (t1/2) was slightly, but not significantly, increased to 2.6 +/- 0.9 hours in patients with severe hepatic impairment (Child-Pugh C), compared with the control group (209 +/ 37 mL/min and 1.8 +/- 0.2 hours, respectively). Liver MR signal enhancement (area under the curve of relative enhancement [%] over time) was similar in patients with mild and moderate hepatic impairment and in those in the control group, but was decreased by 38% in patients with severe hepatic impairment, compared with control. Peak liver enhancement, however, was still at a high level (118% +/- 57%). PK and imaging parameters were not significantly affected in patients with moderate renal impairment (creatinine clearance, 30-50 mL/min). In patients with end-stage renal failure (ESRF), however, the PK profile of Gd-EOB DTPA was significantly different, with an increased t1/2 (20.0 +/- 7.0 hours vs. 1.8 +/- 0.2 hours in the control group). During a 3-hour dialysis session that started 1 hour after administration of the intravenous dose, the serum levels in patients with ESRF declined by between 71% and 88% as a result of elimination by hemodialysis and parallel hepatobiliary excretion. This is comparable with the decline observed in healthy subjects (85%) during the 1- to 4-hour interval after injection. CONCLUSIONS: : The results of the present study show that in humans with moderate renal impairment and mild-to-moderate hepatic impairment, no relevant changes in PK parameters, such as total clearance and t1/2, develop as a result of increased renal excretion to compensate in the case of hepatic impairment (or increased hepatic elimination in the case of renal impairment). The t1/2 of Gd-EOB-DTPA was markedly altered only in patients with ESRF. The high MR signal enhancement profile, observed even in patients with severe hepatic impairment, indicates that there is no need to adjust the dose of Gd-EOB-DTPA. PMID- 21623213 TI - Outcome and prognostic factors in HIV-1-infected patients on dialysis in the cART era: a GESIDA/SEN cohort study. AB - BACKGROUND: Prognosis of HIV-infected patients on dialysis has improved. Few studies have compared survival between HIV-infected and HIV-negative patients on dialysis in the combined antiretroviral therapy (cART) era. We compared the outcome of HIV-infected patients on dialysis with a matched HIV-negative cohort. METHODS: National, multicenter, retrospective cohort study of HIV-infected patients starting dialysis in Spain (1999-2006). Matching criteria for HIV negative patients were dialysis center, year of starting dialysis, age, sex, and race. RESULTS: The study population comprised 122 patients, 66 HIV-infected, and 66 HIV-negative patients. Median age was 41 years, and all but 4 HIV-infected patients were white. HIV-associated nephropathy was only present in 4 cases. HIV infected patients were less frequently included on the kidney transplantation waiting list (17% vs 62%, P < 0.001). They also had more hepatitis C virus coinfection (76% vs 11%, P < 0.001), fewer cardiovascular events (62% vs 88%, P = 0.001), fewer kidney transplants (4.5% vs 38%, P < 0.001), and higher mortality (32% vs 1.5%, P < 0.001). Survival rates [95% confidence interval (CI)] at 1, 3, and 5 years for HIV-infected patients were 95.2% (89.9%-100%), 71.7% (59.7% 83.7%), and 62.7% (46.6%-78.8%). Five-year survival for HIV-negative patients was 94.4% (83.8%-100%) (P < 0.001). Multivariate analysis revealed the following variables to be associated with death in HIV-infected patients: peritoneal dialysis vs hemodialysis [hazard ratio; (95% CI): 2.88 (1.16-7.17)] and being on effective cART [hazard ratio (95% CI): 0.39 (0.16-0.97)]. CONCLUSIONS: Medium term survival of HIV-infected patients on dialysis was lower than that of matched HIV-negative patients. Fewer HIV-infected patients had access to kidney transplantation. Being on effective cART improves survival. Further studies are needed to determine whether peritoneal dialysis increases mortality. PMID- 21623214 TI - Vitamin d status in patients with chronic obstructive pulmonary disease who participate in pulmonary rehabilitation. AB - PURPOSE: Vitamin D deficiency is common in patients with chronic obstructive pulmonary disease; however, no study has evaluated the influence of vitamin D status on effects of pulmonary rehabilitation (PR). METHODS: We studied 311 patients, who participated in a 7-week outpatient PR. Vitamin D status was assessed at entry and examined for association with patient characteristics, dropout from PR, and effect on endurance shuttle walk time. RESULTS: Vitamin D level 25 nM was seen in 61 (19.6%) of the patients. They were significantly younger, were more frequently on long-term oxygen therapy, had higher body mass index and fat-free mass index, had worse quality of life score, tended to have lower percent predicted value for forced expiratory volume in the first second of expiration, and more frequently were current smokers. They had a 3-time higher risk of dropout from the PR program (P = .003) compared with patients with normal vitamin D status and a poorer improvement in endurance shuttle walk time (P = .03). DISCUSSION: In conclusion, vitamin D deficiency was significantly associated with higher dropout rates from PR, and there was a tendency toward a poorer improvement in endurance shuttle walk time. PMID- 21623215 TI - Demographic differences in religious coping after a first-time cardiac event. AB - PURPOSE: The purpose of this project was to describe demographic characteristics of patients who may use religion as a coping response to a first-time cardiac event. METHODS: Patients (N = 105), who were enrolled in cardiac rehabilitation after a first-time myocardial infarction or coronary artery revascularization bypass surgery, completed the Religious Coping Activities Scale. Independent variables included age, gender, religious affiliation, diagnosis, marital status, and education level. The 6 types of religious coping activities were compared for each level of the independent variables. RESULTS: Significant differences emerged for gender, religious affiliation, marital status, and level of education. Women scored higher than men on spiritually based activities (T = 1550, P = .03), good deeds (T = 1504, P = .08), and religious avoidance coping (T = 1505, P = .08). Participants who claimed no religious affiliation scored lowest on good deeds (H[2] = 9.7, P = .008) and interpersonal religious support coping (H[2] = 13.4, P = .001) and higher on discontent coping (H[2] = 5.4, P = .07). Single participants scored higher on spiritually based coping than did married participants (T = 1251, P = .04) and lower on discontent coping (H[1] = 4.3, P = .04). Plead coping was an inverse function of education (H[3] = 6.8, P = .08). CONCLUSIONS: Patients beginning cardiac rehabilitation, particularly those with the demographic characteristics discussed in this study, may benefit from assessment of their desire for pastoral intervention. PMID- 21623216 TI - Patient education and quality of home-based rehabilitation in patients older than 60 years after acute myocardial infarction. AB - PURPOSE: Because only one-third of eligible patients participate in formal cardiac rehabilitation, home-based programs constitute a suitable alternative. We examined effectiveness of a minimal educational intervention on patient fitness and activity levels through the use of simple motivational tools including verbal encouragement and the provision of a booklet containing exercise guidelines and exercise diary. METHODS: We enrolled 186 patients (age, 60 - 78 years; mean age, 69 years; 140 men) who were admitted to the outpatient clinic of Warsaw Institute of Cardiology in 2007-2009 after acute myocardial infarction. Of these, 61.3% had coronary angioplasty with stenting and 30.7% had coronary artery bypass. Patients were randomly assigned into an intervention group receiving minimal educational intervention or control. At baseline and 3 months, assessment was made of cardiopulmonary fitness and autonomic tone with exercise testing. Leisure-time physical activity and atherosclerosis risk factors were assessed at baseline and after 3 and 12 months. RESULTS: At baseline, exercise test results and leisure time activity levels were not significantly different between groups. After 3 months, we noted statistically significant differences in exercise test responses between the intervention group versus control: peak workload 57.3 +/- 2.3 versus 47.2 +/- 2.2 kJ (P < .04) and heart rate recovery 26.5 3.3 versus 23.7 4.2 bpm (P < .001). Leisure-time activity was greater in the intervention group than in control, 3.9 versus 2.3 h/wk (P < .001). Improvement in atherosclerosis risk factors during the course of the study was similar between groups. CONCLUSION: Minimal educational intervention is an effective and safe form of promoting physical activity in older patients after myocardial infarction. PMID- 21623217 TI - Comparison of results of initial trabeculectomy with mitomycin C after prior clear-corneal phacoemulsification to outcomes in phakic eyes. AB - PURPOSE: To compare intraocular pressure (IOP) reduction after trabeculectomy with mitomycin C (MMC) in eyes that had prior clear-corneal phacoemulsification to those in a control group of phakic eyes. METHODS: Thirty-nine pseudophakic eyes (38 patients) that underwent initial trabeculectomy with MMC after clear corneal phacoemulsification between 1997 and 2009 at an academic center were matched on surgeon, age, race, and time of surgery to 39 phakic eyes (39 patients) that underwent the same intervention. Surgical success was defined as: (A) IOP <=18 mm Hg and >=20% reduction of IOP, (B) IOP <=15 mm Hg and >=25% reduction of IOP, and (C) IOP <=12 mm Hg and >=30% reduction of IOP, with or without glaucoma medication. The main outcome measures were success rates, IOP, and number of medications at 1 and 2 years. RESULTS: The median (range) follow-up time was 36 months (1 to 109 mo) and 38 months (1 to 99 mo) in the pseudophakic and phakic groups, respectively. Success rates were not different between the 2 groups (P=0.17, 0.14, and 0.09 for criteria A, B, and C, respectively). The mean (+/-SD) preoperative IOP was 21.4+/-5.9 and 18.6+/-5.5 mm Hg in the pseudophakic and phakic groups, respectively (P=0.034). IOP decreased by 9.3+/-5.6 mm Hg versus 5.7+/-6.9 mm Hg at first year (P=0.057) and by 11.0+/-7.4 mm Hg versus 5.5+/-5.5 mm Hg 2 years after surgery (P=0.008). The number of medications was similar at both follow-up intervals in the 2 groups (P=0.980 and 0.679). CONCLUSIONS: Results of initial trabeculectomy with MMC in eyes with prior clear corneal phacoemulsification are comparable with those in phakic eyes. Clear corneal phacoemulsification does not seem to affect the success rate of subsequent trabeculectomy with MMC. PMID- 21623218 TI - Comparison of the optic nerve imaging by time-domain optical coherence tomography and Fourier-domain optical coherence tomography in distinguishing normal eyes from those with glaucoma. AB - PURPOSE: To assess agreement in cup-to-disc ratio (CDR) estimation between stereoscopic optic disc photography, time-domain optical coherence tomography (OCT), and Fourier-domain OCT, and to compare the discriminating ability of optic nerve head (ONH) parameters by time-domain OCT and Fourier-domain OCT for glaucoma detection. METHODS: Participants underwent stereoscopic photography, Stratus OCT (fast optic disc scan), and RTVue-100 (nerve head mapping), at the same visit. Vertical and horizontal CDRs (VCDR and HCDR) from stereoscopic photography were determined by averaging the results from 2 independent glaucoma specialists. VCDR, HCDR, disc area, cup area, rim area, and cup-to-disc area ratio were acquired from the 2 OCTs. Agreement was assessed by intraclass correlation coefficients (ICCs) and Bland-Altman plots. Area under the receiver operator characteristic curve was used to assess the ability of each parameter to detect glaucoma. RESULTS: A total of 185 eyes were used in final analysis. VCDR and HCDR measurements from RTVue were highest, followed by those from Stratus OCT and stereoscopic photography. CDRs from both OCTs showed excellent agreement with those from stereophotography (ICC, 0.80 to 0.86). Agreement in ONH parameters including CDRs between the 2 OCTs was excellent (ICC, 0.86 to 0.94). There were no statistically significant differences between the ONH parameters from the 2 OCTs with respect to ability to detect glaucoma. CONCLUSIONS: VCDR and HCDR measurement by time-domain and Fourier-domain OCTs exhibited excellent agreement with each other and agreed equally well with those by stereoscopic photography. The ONH parameters between the 2 OCTs performed similarly in determining glaucoma status. PMID- 21623219 TI - Stimulus parameters for multifocal pupillographic objective perimetry. AB - PURPOSE: We compared the diagnostic power of 10 stimulus variants that assessed the visual fields of both eyes by recording pupillary responses to multifocal stimuli. The 10 variants comprised 6 initial tests, and 4 subsequent variants whose design was informed by the initial results. METHODS: Two study groups containing 16 normal and 22 primary open angle glaucoma subjects, and 15 normal and 20 primary open angle glaucoma subjects had their diagnostic status verified by a slit-lamp investigation, applanation tonometry, 3 forms of perimetry, and Stratus OCT. Stereoscopically arranged displays presented multifocal stimulus arrays having 24 stimulus regions/eye within the central 60 degrees. Pupil responses were recorded by video cameras under infrared illumination. The 10 stimulus conditions varied in presentation rate, duration, stimulus luminance, and flicker rate. Stimuli were 4 minutes in duration, presented in 8 segments of 30 seconds. Up to 15% of the data of a segment could be lost owing to blinks and fixation losses without repeating the segment. RESULTS: Each recording gave 96 direct and consensual responses/subject. The best performing stimulus method gave a sensitivity of 1.0 +/- 0.0 (mean +/- SE) for moderate and severe glaucomatous fields combined at a false positive rate of 0.05. Median signal to noise ratios for peak response amplitude expressed as t-statistics exceeded 4 for several variants. CONCLUSIONS: Stimulus delivery rates of about 1 presentation/region/s and test luminance around 150 cd/m performed best diagnostically. Unlike automated perimetry, the mfPOP method provides information on response delays and afferent and efferent defects at each region of the visual field. PMID- 21623220 TI - Outcomes of surgical bleb revision for late-onset bleb leaks after trabeculectomy. AB - PURPOSE: To describe the outcome of surgical bleb revision for late-onset bleb leaks after trabeculectomy. PATIENTS AND METHODS: Appropriate cases were identified. Qualified and complete success required intraocular pressure of 21 mm Hg or less with and without glaucoma medication use, respectively. Bleb survival was determined using Kaplan-Meier survival analysis, and overall success rate was defined as qualified success at last follow-up. Preoperative and postoperative ocular parameters were compared using the signed-rank test. Age, sex, ethnicity, time between leak and revision, and surgeon type (attending vs. surgeons in training) were entered into a logistic regression analysis to assess the impact on surgical outcome. RESULTS: Seventy-eight eyes of 75 patients were included. The overall rate of successful bleb revision was 77%, and qualified and complete success at 24 months was 71% and 34%, respectively. Postoperative complications included early and late bleb leaks in 6% and 9% of the eyes, respectively; bleb related infections in 4% of the eyes; and the need for additional glaucoma surgery in 10% of the eyes. There was no difference in preoperative and postoperative visual acuity (P=0.34) but there was an increase in intraocular pressure (P<0.0001) and the number of medications used (P<0.0001). The number of eyes that did not require glaucoma medication decreased (P=0.002). None of the variables examined had a significant impact on successful surgical outcome. CONCLUSION: Bleb revision showed a high success rate. About two-thirds of eyes required medication, 10% of eyes required additional glaucoma surgery, and there was a low risk for bleb-related infection following bleb revision. PMID- 21623221 TI - Effect of successful and partly successful filtering surgery on the velocity of glaucomatous visual field progression. AB - PURPOSE: We sought to determine whether filtering surgery, even when only partially successful, delays or slows visual field (VF) progression. METHODS: The records of all patients seen in a glaucoma referral practice from 1999 to 2009 were reviewed. Group A comprised eyes with >= 5 VFs before surgery and group B comprised eyes with >= 5 VFs after surgery. Eyes in group B were further divided into those requiring postoperative topical ocular hypotensive therapy (group B-2) and those that did not (group B-1). Automated pointwise linear regression was used to determine global rates (dB/y) of change and progression endpoints. A progression endpoint was determined when 2 or more adjacent test locations in the same hemifield showed a threshold sensitivity decline at a rate of >= 1.0 dB/year with P < 0.01. RESULTS: A total of 206 treated eyes (206 patients; mean age, 63.8 +/- 13.0 y; 11.3 +/- 3.1 VFs; 6.4 +/- 1.8 y follow-up) were included. Mean global VF progression rates in group A (-0.86 +/- 0.8 dB/y) were faster than those in group B (-0.49 +/- 0.9 dB/y, P < 0.01). Group A also had a greater risk of reaching a progression endpoint compared with group B (odds ratio = 2.41, P < 0.01). Groups B-1 and B-2 had different follow-up intraocular pressure means (12.7 +/- 3.7 vs. 15.5 +/- 2.7 mm Hg, respectively; P < 0.01) and peaks (19.4 +/- 5.2 vs. 21.2 +/- 4.2 mm Hg; P = 0.08). The velocity of VF progression was similar for groups B-1 and B-2 (-0.40 +/- 0.6 vs. -0.58 +/- 1.1 dB/y; P = 0.22) and there was no significant difference between the 2 groups regarding the risk of reaching a progression endpoint (odds ratio = 0.83, P = 0.62). CONCLUSIONS: Filtering surgery reduces the rate of disease progression and this effect persists even if adjunctive glaucoma medical therapy is required. PMID- 21623222 TI - Clinical outcomes after lens extraction for visually significant cataract in eyes with primary angle closure. AB - PURPOSE: To examine the clinical outcomes of cataract extraction in eyes with primary angle closure (PAC) and coexisting cataract. PATIENTS AND METHODS: Retrospective study of surgical outcomes after phacoemulsification and intraocular lens implantation in 55 eyes of 39 patients with PAC or occludable angles and visually symptomatic cataract. Approximately, 61.8% of eyes had received a peripheral iridotomy and 65% had evidence of glaucomatous optic neuropathy (GON). RESULTS: At 7.2 months after cataract extraction, a statistically significant reduction in intraocular pressure (IOP) was observed in all eyes with PAC (median 3 mm Hg, P=<0.0001). The reduction in IOP was significantly greater in eyes with a higher preoperative IOP (P=<0.0001). On average, one less glaucoma medication was in use postoperatively (P=0.01). Eyes with >180 degrees of peripheral anterior synechiae (PAS) preoperatively achieved a significantly greater reduction in IOP postoperatively compared with those with less PAS (7.5 vs. 4.4 mm Hg, P=0.03). The observed reduction in IOP in eyes with PAC and GON was significantly greater compared with those without GON (5.6 vs. 2.5 mm Hg, P=0.01). Visual acuity was significantly improved after cataract extraction by a mean of -0.23 LogMAR units (P=0.0001). CONCLUSIONS: Contrary to earlier expectations, the observed reduction in postoperative IOP in eyes with PAC was significantly greater in the presence of a higher preoperative IOP, a larger number of glaucoma medication, narrower iridotrabecular angle width, and greater extent of PAS formation and in eyes with evidence of GON. Lens extraction seems to have a beneficial effect on IOP control in PAC, and is especially efficacious in more advanced cases. PMID- 21623223 TI - Determinants of medication adherence to topical glaucoma therapy. AB - INTRODUCTION/PURPOSE: To determine the associations between medical, demographic, socioeconomic, and ocular factors and adherence to topical glaucoma ocular hypotensive therapy. METHODS: One hundred sixteen patients with ocular hypertension or open-angle glaucoma from 2 tertiary glaucoma services participated in this prospective study. Adherence to ocular hypotensive therapy was measured using an electronic dose monitor (Travatan Dosing Aid, Alcon Laboratories Inc., Fort Worth, TX) and collected data at 3 months after enrollment. We used 3 different definitions of adherence: 1) Definition 1: the proportion of days taking the prescribed number of drops within 3 hours of the prescribed dosing time; 2) Definition 2: the proportion of days taking any drops within 3 hours of the prescribed dosing time; and 3) Definition 3: the proportion of days taking any drops within 6 hours of the prescribed dosing time. Univariate and multivariate models were used to determine the association between the 3 adherence definitions, medical, demographic, socioeconomic, and ocular factors at 3-month follow-up. The main outcome measures for this study were risk factors for poor objective medication adherence. RESULTS: Adherence, using Definition 1, Definition 2, and Definition 3, was 64%, 75%, and 80%, respectively. Age, total number of other eye diseases, and race were significantly associated with full treatment adherence (Definition 1), with race alone significantly predicting 11% of full treatment adherence. For Definition 2, age, income, level of education, and total number of eye diseases were significantly associated with partial adherence (3 h), again race alone significantly predicted 15% of partial adherence (any drops within 3 h). For Definition 3, race, income, level of education, and total number of other eye diseases significantly predicted partial adherence (any drops within 6 h), both race and income predicted 19% of partial treatment adherence. Significant differences for adherence rates between patients of European descent and those of African descent were found for all 3 definitions with those who were less adherent more likely to be of African descent. CONCLUSIONS: Electronic dose monitors provide important information regarding adherence to topical ocular hypotensive medications in glaucoma patients. Electronic dose monitors show low adherence in a significant number of participants. Future studies are needed to determine the reasons for these differences in health behaviors related to glaucoma treatment, which should guide treatment of poor adherence with glaucoma therapy. PMID- 21623224 TI - Clinical characterization of young chinese myopes with optic nerve and visual field changes resembling glaucoma. AB - PURPOSE: To clinically characterize young Chinese myopic patients with optic disc changes suggestive of glaucoma. METHODS: In this prospective study, patients <=50 years of age who had myopia and suspected glaucomatous optic neuropathy (group A) were compared with established glaucoma patients (group B) and normals (group C). Risk factors such as family history of glaucoma and history of vasospasm and ischemic risk factors were compared. Ophthalmic examination included optic disc assessment and static automated perimetry. Peak diurnal intraocular pressure (IOP) and fluctuation were measured for groups A and C. RESULTS: Ninety-two patients were enrolled (32 patients in group A, 43 in group B, and 21 in group C). Mean refractive error differed between group A [-6.59 Diopter sphere (DS); 95% CI, -8.51 to -4.68] and group B (-0.92 DS; 95% CI, -2.05 to 0.19) (P<0.0001) and C (-0.17 DS; 95% CI, -0.48 to 0.14) (P<0.0001). Optic disc tilt was more common in group A (31.3%) compared with group B (16.3%, P=0.01); both groups had comparable vertical cup-to-disc ratios (P=0.58). The visual field mean pattern standard deviation was higher for group A compared with group B (P=0.03), whereas mean deviation was similar (P=0.42); group A had less frequent hemifield defects (P=0.02). Mean peak diurnal IOP in group A (19.0 mm Hg; 95% CI, 17.3-20.7) was significantly higher than group C (15.8 mm Hg; 95% CI, 14.6-17.0, P=0.007); mean IOP fluctuation showed no difference (P=0.19). Ischemic risk factors were more common in group B compared with A (P<0.0001). CONCLUSIONS: Although the clinical features of myopia and glaucoma-like optic nerve changes in young Chinese individuals may allow differentiation from older glaucoma patients of the same ethnicity, such a distinction is not reliable enough to make definitive differential management recommendations for these groups in all circumstances at the present time. A prospective observational study with matching of groups on potentially confounding variables is necessary to better assess and compare the natural history of glaucomatous disease in such groups of patients. PMID- 21623225 TI - Glaucoma progression after the first-detected optic disc hemorrhage by optical coherence tomography. AB - PURPOSE: To evaluate the progression of glaucoma after first detected disc hemorrhage (DH) by time-domain optical coherence tomography (OCT). METHODS: Forty four eyes with a unilateral DH were allocated to the DH group and 44 fellow eyes to the non-DH group. OCT signal decrease indicating retinal nerve fiber layer thinning was defined as a decrease beyond the upper 95% limit of test-retest variability defined at the 95% confidence level of 44 control patients. The DH and non-DH groups were compared in terms of OCT-derived glaucoma progressions after the first detected DH. In addition, clock-hour locations of the OCT-derived progression were compared with those of DHs. RESULTS: After a median 38.8+/-10.8 months of follow-up, 72.7% (32 of 44) of eyes in the DH group showed OCT determined progression on a clock-hour basis, compared with 27.3% (12 of 44) of the fellow eyes in the non-DH group (P<0.0001, Fisher exact test). Eyes in the DH group showed more rapid and greater falls in clock-hour-based progression than fellow eyes in the non-DH group (P<0.0001, log-rank test). Twenty-nine eyes (90.6%) showed progression within 1 clock hour of the location of DH. However, no significant intergroup differences were observed in terms of OCT-determined progression on quadrant or average basis (P>0.05, respectively, Fisher exact test). CONCLUSIONS: Rapid, spatially compatible, localized thinning of the retinal nerve fiber layer after the first detected DH was shown by Stratus OCT. PMID- 21623226 TI - Myopic optic disc tilt and the characteristics of peripapillary retinal nerve fiber layer thickness measured by spectral-domain optical coherence tomography. AB - PURPOSE: To investigate the correlation between myopic optic disc tilt and the characteristics of peripapillary retinal nerve fiber layer (RNFL) thickness measured by Cirrus HD spectral-domain optical coherence tomography (Cirrus HD OCT; Carl Zeiss Meditec, Dublin, CA). METHODS: A total of 255 eyes of 255 healthy young male participants with various degrees of refractive errors (mean spherical equivalent, -3.17 +/- 2.40 D; range, -11.00 to 0.00 D) underwent ophthalmic examinations, including refractive error, axial length, and optic disc area measurement. The degree of horizontal/vertical optic disc tilt was evaluated by cross-sectional images obtained by the Cirrus HD OCT. The average, superior, nasal, inferior, and temporal quadrant thickness and superior/inferior peak locations of the peripapillary RNFL were also measured with the Cirrus HD OCT. RESULTS: On the univariate analysis, eyes with more temporally tilted optic discs (horizontal tilt) had higher myopia, greater axial length, a thinner average, superior, nasal, and inferior RNFL, thicker temporal RNFL, and more temporally positioned superior/inferior peak locations (all P values <0.001). The degree of inferior optic disc tilt (vertical tilt) was associated with high myopia and a more temporally positioned inferior peak location (all P values <0.05). On multivariate analysis, eyes with more temporally tilted optic discs had a thicker temporal RNFL and more temporally positioned superior/inferior peak locations. CONCLUSIONS: The characteristics of the peripapillary RNFL thickness were associated with the degree of myopic optic disc tilt, especially in the temporal area. The degree of myopic optic disc tilt should be considered when interpreting the RNFL thickness measured by the Cirrus HD OCT. PMID- 21623227 TI - Peripheral homonymous hemianopia: correlation between lesion location and visual field defects by means of cytoarchitectonic probabilistic maps. AB - BACKGROUND: Peripheral homonymous scotomas beyond 30 degrees from fixation are rare. The paucity of publications describing such visual field defects might be attributed to various factors, including the absence of severe symptoms, routine visual field assessment restricted to the central 30 degrees with automated perimetry, and the collateral circulation to the occipital cortex. The aim of this study was to correlate the brain lesions and perimetric findings in 2 unusual cases of peripheral homonymous scotomas, with the anatomic location of the optic radiation and primary visual cortex. METHODS: Two patients with circumscribed homonymous scotomas beyond 30 degrees related to infarcts in the intermediate area of the visual cortex are reported. We describe a new strategy, which relies on modern lesion analysis and stereotaxic probabilistic cytoarchitectonic maps, to accurately correlate the brain lesion site with the location of the peripheral homonymous visual field defects. RESULTS: In Case 1, the posterior optic radiation was affected in its termination in the upper intermediate visual cortex. In Case 2, the lesion was located in the upper rostral portion of the primary visual cortex. In both, the most anterior part of the visual cortex and the occipital pole were intact, accounting for preservation of the central and most peripheral visual field. Additionally, correlation of the neuroimaging findings with commonly used maps of the representation of the visual field on the striate cortex suggested that our data were most consistent with the Holmes map. CONCLUSIONS: Modern lesion analysis and cytoarchitectonic maps, in combination with the existing retinotopic maps, may provide reliable clues for the localization of cerebral infarction and prognosis of homonymous visual field defects and may lead to a better understanding of the link between neuroanatomical landmarks and functional outcomes. PMID- 21623228 TI - Neuro-ophthalmological features of primary diffuse leptomeningeal gliomatosis. AB - We performed an in-depth study of the neuro-ophthalmologic signs and symptoms of a rare but fatal disease known as primary diffuse leptomeningeal gliomatosis (PDLG). Two new cases of PDLG are described, and 22 published cases reviewed. Papilledema and sixth nerve palsy are the most common neuro-ophthalmic findings. Other abnormalities include third and fourth nerve palsies, nystagmus, and vision loss. Involvement of the visual system may be part of the initial presentation of PDLG. PMID- 21623229 TI - MRI restricted diffusion in lymphomatous optic neuropathy. AB - Restricted diffusion in the optic nerve detected with MRI has been previously reported in infarction and inflammation but not in infiltrative neoplasm. We report a 44-year-old man with recently diagnosed non-Hodgkin B-cell lymphoma who developed an acute left optic neuropathy. MRI showed no evidence of brain parenchymal or meningeal lymphoma but did show restricted diffusion in the intraorbital portion of the affected optic nerve. Despite treatment with corticosteroid, standard chemotherapy, and orbital X-irradiation, visual function did not improve. The restricted diffusion persisted on a follow-up MRI performed 4 months after the onset, a phenomenon that is atypical for infarction. Perhaps, this persisting imaging abnormality in lymphomatous optic neuropathy reflects the dense cellularity of the neoplasm. PMID- 21623230 TI - Ocular oscillations in the neuromyelitis optica spectrum. AB - Four French West Indian women complained of oscillopsia and were found to have an acquired eye movement disorder. In 3 of them, different types of nystagmus were found, including upbeat, downbeat, and central form of vestibular nystagmus. One developed opsoclonus-myoclonus syndrome. Three patients had neuromyelitis optica antibodies, and 3 had brainstem abnormalities detected on MRI. Two patients had definite NMO, while the other 2 were considered to be at high risk for developing NMO. Treatment with high-dose systemic corticosteroids, with plasma exchanges, or in combination led to resolution of oscillopsia. We propose that eye movement disorders be added to the neurological manifestations of NMO. PMID- 21623231 TI - Clevidipine for controlled hypotension during spinal surgery in adolescents. AB - BACKGROUND: Clevidipine is an ultrashort-acting, intravenous calcium channel antagonist. Metabolism by blood and tissue esterases results in a half-life of 1 to 2 minutes. We present preliminary experience with this novel agent to provide controlled hypotension (CH) in a cohort of adolescents undergoing spinal surgery. METHODS: The records of patients <=18 years of age who received clevidipine for CH were retrospectively reviewed. Demographic data included age, weight, sex, and comorbid disease processes. Information regarding clevidipine included the initial infusion rate, time to achieve the target mean arterial pressure (MAP), the maintenance infusion rate, the average infusion rate, and the duration of administration. Hemodynamic information included the starting MAP and heart rate (HR) and the MAP and HR during the clevidipine infusion. Charts were reviewed for adverse effects related to clevidipine including excessive hypotension (MAP <=50 mm Hg, the need to discontinue the infusion or the need for a fluid bolus or administration of a vasopressor) or the administration of a beta-adrenergic antagonist. RESULTS: The study cohort included 20 patients, ranging in age from 14 to 18 years and in weight from 46 to 96 kg. A total intravenous anesthetic technique (propofol, remifentanil, and dexmedetomidine) was used. When the MAP was >=65 mm Hg, clevidipine was added to maintain the MAP at 50 to 65 mm Hg. The clevidipine infusion was started at 0.5 to 1 MUg/kg/min, and increased in increments of 0.5 to 1 MUg/kg/min every 2 to 3 minutes to achieve the desired MAP. The target MAP was achieved within 5 minutes in 15 of the 20 patients and within 10 minutes in the other 5 patients. The maintenance infusion rate of clevidipine varied from 1 to 5 MUg/kg/min (2.9+/-0.7 MUg/kg/min). With the administration of clevidipine, HR increased from a baseline of 76+/-14 to 92+/-11 beats/min (P<0.05). The HR increase was >=20 beats/min in 4 patients. Intermittent doses of metoprolol were administered to 3 patients. No excessive hypotension was noted. When the clevidipine infusion was discontinued, MAP returned to baseline within 5 minutes in 16 of the 20 patients and within 10 minutes in the other 4 patients. CONCLUSIONS: Clevidipine maintained the MAP at 50 to 65 mm Hg and provided CH. Mild tachycardia was noted in some patients with the occasional need for a beta-adrenergic antagonist. No episodes of excessive hypotension were noted. Given its short half-life, clevidipine can be rapidly titrated to provide CH when changing levels of sympathetic stimulation may occur. PMID- 21623233 TI - Evaluation of emergency department evidence-based practices to prevent the incidence of ventilator-acquired pneumonia. AB - Pathogens in the oropharynx may be transported to the lung parenchyma during intubation and cause ventilator-acquired pneumonia. This project evaluated 3 post intubation evidence-based practices in the emergency department: oral care, head of-bed elevation, and suctioning above the endotracheal tube balloon. Ventilator acquired pneumonia cases decreased 83% after implementing these practices. There is no need to wait for the patient to be admitted to intensive care to begin these evidence-based practices. PMID- 21623234 TI - Gastric adenocarcinoma presenting as thrombotic microangiopathy in a 14-year-old girl. AB - Gastric adenocarcinoma is rare in childhood and often presents with disseminated malignancy at diagnosis due to aspecific symptoms leading to delay in diagnosis. A familial predisposition for gastrointestinal cancer is suggested for the development of this early-onset adenocarcinoma. We report the case of a 14-year old girl with a familial history of colorectal, liver, and breast cancers affected by metastatic gastric adenocarcinoma, who first presented with thrombotic microangiopathy. Thrombotic microangiopathy as first clinical presentation of metastatic gastric cancer is an exceptional event in childhood and represents a challenge for pediatricians. Gastric adenocarcinoma should be suspected in young patients with a significant familial history and also in the absence of initial specific signs, so as to provide correct diagnosis and appropriate treatment. PMID- 21623235 TI - Computed tomography findings predicting invasiveness of thymoma. AB - PURPOSE: To identify preoperative computed tomography (CT) findings associated with thymoma invasiveness before surgical resection and with clinical outcome. METHODS: We retrospectively reviewed CT scans of 99 patients with thymoma surgically treated at our institution between September 1999 and April 2010. Chest CT findings documented were size, volume, and heterogeneity of primary tumor; abutment of mediastinal vessels; and presence of calcifications, lobulation, infiltration of fat surrounding tumor, adjacent pulmonary changes, adenopathy, and pleural nodularity. RESULTS: Our study group consisted of 53 (54%) men and 46 (46%) women, age 18-79 years (mean: 53.2 years). Masaoka pathologic stages were stage I for 10 (10%), stage II for 48 (48%), stage III for 21 (21%), and stage IV for 20 (20%). The median radiologic tumor size was 7 cm (range: 2.5-21 cm). A multivariable logistic regression model showed that primary tumors with prechemotherapy radiologic tumor size >= 7 cm (odds ratio [OR]: 3.18, 95% confidence interval [CI]: 1.16-8.67, p = 0.02), a lobulated tumor contour (OR: 8.20, 95% CI: 1.63-41.35, p = 0.01), and infiltration of surrounding fat (OR: 3.76, 95% CI: 1.45-9.78, p = 0.007) were more likely to have stage III or IV disease. Cox's proportional hazard model showed that the presence of pulmonary nodules on staging CT was the only imaging parameter associated with shorter progression-free survival (hazard ratio: 4.93, 95% CI: 1.60-15.17, p = 0.005) and overall survival (p = 0.03). CONCLUSION: The primary tumor CT imaging features can differentiate between stage I/II and stage III/IV disease and, thus, help identify patients more likely to benefit from neoadjuvant therapy. PMID- 21623236 TI - Utility of 10 immunohistochemical markers including novel markers (desmocollin-3, glypican 3, S100A2, S100A7, and Sox-2) for differential diagnosis of squamous cell carcinoma from adenocarcinoma of the Lung. AB - INTRODUCTION: Recent clinical trials revealed that accurate histologic typing of non-small cell lung cancer, especially squamous cell carcinoma (SCC), is essential. PATIENTS AND METHODS: We analyzed 10 antibodies expression in 150 SCC cases (53 well-, 51 moderately, and 46 poorly differentiated cases) and 159 adenocarcinoma (AC) cases (49 well-, 52 moderately, and 58 poorly differentiated cases). RESULTS: In all SCC and AC cases, p63 was the most sensitive marker for SCC (98.7%), followed by high-molecular-weight (HM) cytokeratin (CK) (97.3%), CK5/6 (93.3%), Sox2 (80%), thrombomodulin (79.3%), desmocollin-3 (72.7%), S100A7 (70.7%), S100A2 (63.3%), and glypican-3 (46.7%). Desmocollin-3 was the most specific marker for SCC (100%), followed by CK5/6 (98%), Sox2 (95.5%), glypican-3 (92.4%), S100A7 (86.8%), thrombomodulin (79.9%), S100A2 (64.6%), p63 (51.6%), and HMCK (33.3%). Thyroid transcription factor-1 (TTF-1) expression was observed in 87.4% of AC cases and 2.0% of SCC cases. When analyzing only poorly differentiated tumors, HMCK was the most sensitive marker for SCC (100%), followed by p63 (97.8%), CK5/6 (87.0%), Sox2 (71.7%), thrombomodulin (58.7%), desmocollin-3 (52.2%), S100A2 (50%), glypican-3 (45.7%), and S100A7 (45.7%). Desmocollin-3 was the most specific marker for poorly differentiated SCC (100%), followed by CK5/6 (98.3%), glypican-3 (94.8%), Sox2 (94.8%), S100A2 (81%), S100A7 (75.9%), thrombomodulin (72.4%), p63 (48.3%), and HMCK (36.8%). The 10-fold crossvalidated classification and regression tree analysis revealed that the combination of CK5/6 and TTF-1 was the best immunohistochemical marker panel for the differentiation between SCC and AC. CONCLUSION: CK5/6 is the best marker for differentiating SCC and AC, irrespective of the histological grade of the tumor. Thus, the combination of CK5/6 and TTF-1 is the most recommended combination of immunohistochemical markers. PMID- 21623237 TI - Radiological changes after stereotactic radiotherapy for stage I lung cancer. AB - INTRODUCTION: Stereotactic body radiation therapy (SBRT) is entering routine clinical use for selected patients with early-stage non-small cell lung cancer. Post-SBRT radiological changes are commonly seen on follow-up computed tomography (CT) imaging and can cause diagnostic dilemmas. The aim of this study is to describe the incidence, radiological severity, and long-term morphology of these changes. METHODS: CT scans from patients treated between 2003 and June 2008 were eligible for evaluation if radiological follow-up had been performed at our center for at least 2 years, and there was no definite evidence of local recurrence. Timing, incidence, morphology, and severity of lung changes were determined. RESULTS: CT scans from 61 patients (68 lesions) with a median follow up of 2.5 years were evaluated. Within 6 months, 54% of lesions were associated with additional radiological abnormalities, and this figure reached 99% after 36 months. Most changes were scored as mild to moderate, and although the median time to first observation was 17 weeks, 25% appeared >= 1 year post-SBRT. In 47% of lesions, the morphology or severity of changes continued to evolve more than 2 years posttreatment. CONCLUSIONS: Mild-moderate radiological changes are common after lung SBRT. Some degree of late change is nearly universal, and it often continues to evolve more than 2 years post-SBRT. Clinicians should be aware of these radiological findings, which need to be distinguished from the uncommon cases of local failure post-SBRT. PMID- 21623238 TI - CXCR4 overexpression is associated with poor outcome in females diagnosed with stage IV non-small cell lung cancer. AB - BACKGROUND: It has been proposed that the chemokine receptor, CXCR4, and its ligand, stromal cell-derived factor-1 (SDF-1), play a critical role in organ specific tumor metastasis. High CXCR4 expression in resected non-small cell lung cancer (NSCLC) tumors is associated with poorer outcome; however, its effect on patient outcome in advanced NSCLC has not been explored. METHODS: After institutional ethical approval was obtained, demographic details, clinical variables, and outcome data were collected on consecutive NSCLC patients diagnosed at the Tom Baker Cancer Centre from 2003 to 2006 (Glans-Look Lung Cancer Database). Formalin-fixed paraffin-embedded diagnostic biopsies from stage IV patients were obtained and tissue microarrays generated. CXCR4 expression within NSCLC cells was analyzed by quantitative fluorescent immunohistochemistry using the HistoRx PM-2000 platform and then correlated with clinical outcome. RESULTS: Of 832 patients, 170 had samples suitable for tissue microarray generation and analysis. Automated immunohistochemistry for CXCR4 was successfully completed on all 170 patients. High expressors had a significantly poorer median overall survival of 2.7 months versus 5.6 months for the low expressors (p = 0.0468). This difference is driven by high-expressing females who have a median overall survival of 1.6 months versus 6.4 months for the low expressors (p = 0.006). CONCLUSIONS: CXCR4 is expressed in the majority of NSCLC tumors, and overexpression is associated with significantly poorer survival in stage IV NSCLC patients. Interestingly, this poor outcome is disproportionately represented in the female population. Our results suggest a gender-dependent difference in clinical outcome based on CXCR4 overexpression in stage IV NSCLC. PMID- 21623239 TI - The risk of cytotoxic chemotherapy-related exacerbation of interstitial lung disease with lung cancer. AB - INTRODUCTION: It is unknown what type of interstitial lung disease (ILD) has high risk for chemotherapy-related exacerbation of ILD. We investigated the risk of exacerbation of ILD for patients with lung cancer with ILD. METHODS: One hundred nine patients with lung cancer with ILD treated with cytotoxic chemotherapy at Shizuoka Cancer Center between August 2002 and April 2010 were retrospectively reviewed. RESULTS: On pretreatment computed tomography (CT) of the chest, 69 patients (63%) were identified with usual interstitial pneumonia (UIP) pattern, and 40 patients (37%) had non-UIP pattern. Patients with UIP pattern developed cytotoxic chemotherapy-related exacerbation of ILD more frequently than those with non-UIP pattern (30 versus 8%, p = 0.005). The incidence of grade 5 pulmonary toxicities was 9% in patients with UIP pattern, compared with 3% in those with non-UIP pattern. Multivariate analyses demonstrated that age (<70 years) and CT pattern (UIP) were significant independent risk factors for cytotoxic chemotherapy-related exacerbation of ILD. In small cell lung cancer, overall survival (OS) from the start of first-line chemotherapy was significantly shorter in UIP pattern than non-UIP pattern (median OS: 9 versus 16 months, p = 0.0475), whereas there was no significant difference in patients with non-small cell lung cancer (median OS: 12 versus 9 months, p = 0.2529). CONCLUSIONS: Our results indicated that the incidence of exacerbation of ILD was significantly higher in patients with lung cancer with UIP pattern on CT findings than in those with non-UIP pattern. Therefore, great care is required when administering cytotoxic chemotherapy agents for patients with lung cancer with UIP pattern. PMID- 21623240 TI - A budget impact analysis of telemedicine-based collaborative care for depression. AB - BACKGROUND: Patients with depression use more health services than patients without depression. However, when depression symptoms respond to treatment, use of health services declines. Most depression quality improvement studies increase total cost in the short run, which if unevenly distributed across stakeholders, could compromise buy-in and sustainability. The objective of this budget impact analysis was to examine patterns of utilization and cost associated with telemedicine-based collaborative care, an intervention that targets patients treated in small rural primary care clinics. METHODS: Patients with depression were recruited from VA Community-based Outpatient Clinics, and 395 patients were enrolled and randomized to telemedicine-based collaborative care or usual care. Dependent variables representing utilization and cost were collected from administrative data. Independent variables representing clinical casemix were collected from self-report at baseline. RESULTS: There were no significant group differences in the total number or cost of primary care encounters. However, as intended, patients in the intervention group had significantly greater depression related primary care encounters (marginal effect=0.34, P=0.004) and cost (marginal effect=$61.4, P=0.013) to adjust antidepressant therapy for nonresponders. There were no significant group differences in total mental health encounters or cost. However, as intended, the intervention group had significantly higher depression-related mental health costs (marginal effect=$107.55, P=0.03) due to referrals of treatment-resistant patients. Unexpectedly, patients in the intervention group had significantly greater specialty physical health encounters (marginal effect =0.42, P=0.001) and cost (marginal effect =$490.6, P=0.003), but not depression-related encounters or cost. Overall, intervention patients had a significantly greater total outpatient cost compared with usual care (marginal effect=$599.28, P=0.012). CONCLUSIONS: Results suggest that telemedicine-based collaborative care does not increase total workload for primary care or mental health providers. Thus, there is no disincentive for mental health providers to offer telemedicine-based collaborative care or for primary care providers to refer patients to telemedicine-based collaborative care. PMID- 21623241 TI - Emerging perspectives on transforming the healthcare system: key conceptual issues. AB - BACKGROUND: Despite reform efforts, today's healthcare system faces multiple challenges. Limitations of the current system result in major gaps between evidence and practice, suboptimal quality, inequitable patterns of utilization, inadequate safety and reliability, and unsustainable cost increases. Furthermore, there are major problems of overuse, underuse, and misuse of healthcare. Beyond the healthcare reform legislation passed in 2010 mandate, major transformation of how U.S. A healthcare system is comprised of multiple levels. PURPOSE: This article briefly outlines the need for transformational change in healthcare and key conceptual issues. METHODS: We identified literature and concepts relevant to developing a research agenda on transformational change in healthcare. Relevant concepts were further developed through conference presentations and discussions. CONCLUSIONS: This introduction to the 10th Regenstrief Conference proceedings provides a foundation for understanding key terms, questions, and concepts relevant to the area of transformational change in healthcare. PMID- 21623242 TI - Quantification of glioma removal by intraoperative high-field magnetic resonance imaging: an update. AB - BACKGROUND: The beneficial role of the extent of resection (EOR) in glioma surgery in correlation to increased survival remains controversial. However, common literature favors maximum EOR with preservation of neurological function, which is shown to be associated with a significantly improved outcome. OBJECTIVE: In order to obtain a maximum EOR, it was examined whether high-field intraoperative magnetic resonance imaging (iMRI) combined with multimodal navigation contributes to a significantly improved EOR in glioma surgery. METHODS: Two hundred ninety-three glioma patients underwent craniotomy and tumor resection with the aid of intraoperative 1.5 T MRI and integrated multimodal navigation. In cases of remnant tumor, an update of navigation was performed with intraoperative images. Tumor volume was quantified pre- and intraoperatively by segmentation of T2 abnormality in low-grade and contrast enhancement in high grade gliomas. RESULTS: In 25.9% of all cases examined, additional tumor mass was removed as a result of iMRI. This led to complete tumor resection in 20 cases, increasing the rate of gross-total removal from 31.7% to 38.6%. In 56 patients, additional but incomplete resection was performed because of the close location to eloquent brain areas. Volumetric analysis showed a significantly (P < .01) reduced mean percentage of tumor volume following additional further resection after iMRI from 33.5% +/- 25.1% to 14.7% +/- 23.3% (World Health Organization [WHO] grade I, 32.8% +/- 21.9% to 6.1% +/- 18.8%; WHO grade II, 24.4% +/- 25.1% to 10.8% +/- 11.0%; WHO grade III, 35.1% +/- 27.3% to 24.8% +/- 26.3%; WHO grade IV, 34.2% +/- 23.7% to 1.2% +/- 16.2%). CONCLUSION: MRI in conjunction with multimodal navigation and an intraoperative updating procedure enlarges tumor volume reduction in glioma surgery significantly without higher postoperative morbidity. PMID- 21623243 TI - Spinal subarachnoid hemorrhage mimicking a carotid artery dissection. PMID- 21623245 TI - Prognostic models for pediatric head trauma still in their infancy. PMID- 21623246 TI - Detection of pulsation in ruptured and unruptured cerebral aneurysms by electrocardiographically gated 3-dimensional computed tomographic angiography with a 320-row area detector computed tomography and evaluation of its clinical usefulness. AB - BACKGROUND: In ruptured cerebral aneurysms (RCAs), identification of the rupture point of a cerebral aneurysm is useful for treatment planning. In unruptured cerebral aneurysms (URCAs), detection of the risk of aneurysmal rupture is also useful for patient management. OBJECTIVE: Electrocardiographic (ECG)-gated 3D-CT angiography was performed for patients with RCAs and URCAs using 320-row area detector CT (ADCT) to detect pulsation of the cerebral aneurysms. The clinical usefulness of this method was then evaluated. METHODS: Twelve patients had 12 RCAs, and 39 patients had 53 URCAs. A 320-row ADCT system was used to scan. ECG gated reconstruction was then performed with the R-R interval divided into 20 phases. RESULTS: Pulsation was observed in 10 of the 12 RCAs. The bleeding site was considered to correspond to the area of pulsation. Pulsation was observed in 14 of 53 URCAs. Thirteen patients with 18 URCAs were followed. Of the 11 URCAs in which pulsation was not observed, 1 showed a change in shape. Of the 7 URCAs in which pulsation was observed, 3 showed a change in shape. URCAs in which pulsation was observed were more likely to show a change in shape (P = .082). CONCLUSION: The area of pulsation was found to correspond to the bleeding site in many RCAs. This information would be extremely useful for treatment planning. The detection of pulsation in an URCA is therefore considered to provide useful information for patient management. PMID- 21623247 TI - Results of embolization used as the first treatment choice in a consecutive nonselected population of ruptured aneurysms: clinical results of the Clarity GDC study. AB - BACKGROUND: The International Subarachnoid Aneurysm Trial (ISAT) showed that for ruptured aneurysms suitable for both techniques, coiling should be the first choice treatment. Only a small proportion of patients (22%) with ruptured aneurysms were included in that trial. Operators were selected on their experience. One could then criticize the impact of the ISAT on clinical practice as a result of recruitment biases and operators' selection. OBJECTIVE: To evaluate the morbidity and mortality of coiling when used as first-choice treatment in a consecutive population of patients with ruptured aneurysms treated by nonselected operators. METHODS: Thirty-four operators from 19 French centers treated 405 patients with GDC coils from November 2006 to July 2007. The method of treatment was not prespecified. RESULTS: World Federation of Neurological Societies grade at admission was I/II in 65.7% and IV/V in 30.6% of patients. At the 3- to 6-month follow-up, 23.3% of patients were dependent or dead. Thromboembolic events and intraoperative rupture resulted in permanent deficit in 13 (3.2%) and 2 (0.5%), respectively, and death in 4 (1.0%) and 0. Early rebleeding occurred in 2 patients (0.5%) with 2 subsequent deaths. Permanent treatment morbidity and mortality were 3.7 % and 1.5 %, respectively. CONCLUSION: Clinical results of the multicenter prospective Clarity registry show that when coiling is performed as first-intention treatment in a consecutive series of nonselected ruptured aneurysms by nonselected operators, clinical results are similar to those of the ISAT. PMID- 21623248 TI - Tubular diskectomy vs conventional microdiskectomy for the treatment of lumbar disk-related sciatica: cost utility analysis alongside a double-blind randomized controlled trial. AB - BACKGROUND: Conventional microdiskectomy is the most frequently performed surgery for patients with sciatica caused by lumbar disk herniation. Transmuscular tubular diskectomy has been introduced to increase the rate of recovery, although evidence of its efficacy is lacking. OBJECTIVE: To determine whether a favorable cost-effectiveness for tubular diskectomy compared with conventional microdiskectomy is attained. METHODS: Cost utility analysis was performed alongside a double-blind randomized controlled trial conducted among 325 patients with lumbar disk related sciatica lasting >6 to 8 weeks at 7 Dutch hospitals comparing tubular diskectomy with conventional microdiskectomy. Main outcome measures were quality-adjusted life-years at 1 year and societal costs, estimated from patient reported utilities (US and Netherlands EuroQol, Short Form Health Survey-6D, and Visual Analog Scale) and diaries on costs (health care, patient costs, and productivity). RESULTS: Quality-adjusted life-years during all 4 quarters and according to all utility measures were not statistically different between tubular diskectomy and conventional microdiskectomy (difference for US EuroQol, -0.012; 95% confidence interval, -0.046 to 0.021). From the healthcare perspective, tubular diskectomy resulted in nonsignificantly higher costs (difference US $460; 95% confidence interval, -243 to 1163). From the societal perspective, a nonsignificant difference of US $1491 (95% confidence interval, 1335 to 4318) in favor of conventional microdiskectomy was found. The nonsignificant differences in costs and quality-adjusted life-years in favor of conventional microdiskectomy result in a low probability that tubular diskectomy is more cost-effective than conventional microdiskectomy. CONCLUSION: Tubular diskectomy is unlikely to be cost-effective compared with conventional microdiskectomy. PMID- 21623249 TI - Determining spherical lens correction for astronaut training underwater. AB - PURPOSE: To develop a model that will accurately predict the distance spherical lens correction needed to be worn by National Aeronautics and Space Administration astronauts while training underwater. The replica space suit's helmet contains curved visors that induce refractive power when submersed in water. METHODS: Anterior surface powers and thicknesses were measured for the helmet's protective and inside visors. The impact of each visor on the helmet's refractive power in water was analyzed using thick lens calculations and Zemax optical design software. Using geometrical optics approximations, a model was developed to determine the optimal distance spherical power needed to be worn underwater based on the helmet's total induced spherical power underwater and the astronaut's manifest spectacle plane correction in air. The validity of the model was tested using data from both eyes of 10 astronauts who trained underwater. RESULTS: The helmet's visors induced a total power of -2.737 D when placed underwater. The required underwater spherical correction (FW) was linearly related to the spectacle plane spherical correction in air (FAir): FW = FAir + 2.356 D. The mean magnitude of the difference between the actual correction worn underwater and the calculated underwater correction was 0.20 +/- 0.11 D. The actual and calculated values were highly correlated (r = 0.971) with 70% of eyes having a difference in magnitude of <0.25 D between values. CONCLUSIONS: We devised a model to calculate the spherical spectacle lens correction needed to be worn underwater by National Aeronautics and Space Administration astronauts. The model accurately predicts the actual values worn underwater and can be applied (more generally) to determine a suitable spectacle lens correction to be worn behind other types of masks when submerged underwater. PMID- 21623250 TI - Ten-year changes in fusional vergence, phoria, and nearpoint of convergence in myopic children. AB - PURPOSE: To identify longitudinal changes in fusional vergence ranges and their relationship to other clinical measures in young myopic subjects. METHODS: Measurements were collected annually for 10 years on 114 subjects from the University of Houston Correction of Myopia Evaluation Trial cohort. Subject age was 7 to 13 years at year 1 of follow-up. Measurements included refractive error, distance and near phoria, interpupillary distance (IPD), prism bar fusional vergence ranges, and nearpoint of convergence (NPC). Multilevel modeling was used to determine baseline and rate of change for fusional vergence ranges and the impact of phoria, IPD, and NPC on these measures. RESULTS: Year 1 mean distance base-out (BO) break was 20 prism diopters (pd) and decreased 5.6 pd over 10 years (p < 0.001). Mean near BO break was 30 pd at year 1 and decreased 9.4 pd over 10 years (p < 0.001). Greater esophoria was significantly related to greater BO break (p < 0.02) and receded NPC was significantly related to lower magnitude BO break at near (p < 0.001). Distance IPD increased 3 mm over 10 years (p < 0.001) but was unrelated to the magnitude of the BO ranges (p > 0.2). Mean distance base in (BI) break was 7 pd at year 1 and increased 0.5 pd in 10 years (p = 0.04). Mean near BI break was 13 pd at year 1 and did not significantly change. Mean distance phoria was 0.1 pd exophoria at year 1 and did not change, whereas near phoria was 2.4 pd esophoria at year 1 and became more exophoric (4 pd in 10 years, p < 0.001). CONCLUSIONS: These results suggest that for myopic children convergence ranges decrease for both distance and near viewing during the school years as near phoria becomes more exophoric. These findings could have clinical implications given that compensating convergence ranges decrease as near phoria becomes more divergent. PMID- 21623251 TI - Characteristics of astigmatism in a population of schoolchildren, Dezful, Iran. AB - PURPOSE: To study and analyze astigmatism and its characteristics in a cross sectional study of schoolchildren from Dezful, Iran. METHODS: In a cross sectional study using random cluster sampling on 460 schools in Dezful (clusters), 39 clusters were selected. The study was conducted after coordinating with schools and obtaining written consent from students' parents. Cycloplegic refraction was done for primary and junior high school students and non cycloplegic refraction was done for high school students. Astigmatism was defined as the cylinder power of 0.75 diopter (D) or more. RESULTS: Of 5726 selected students, 5544 (96.8%) participated in the study. The prevalence of astigmatism was 13.47% (95% confidence interval: 11.90 to 15.04) and was not significantly related to age and gender. Regarding axis, 45.76, 48.14, and 6.09% of astigmatic schoolchildren had with-the-rule (WTR), against-the-rule (ATR), and oblique astigmatism, respectively. An increase in age was accompanied by a decrease in the prevalence of WTR astigmatism and an increase in the prevalence of ATR astigmatism (p < 0.001). The association between astigmatism and myopia [odds ratio = 8.81] was stronger than its association with hyperopia (odds ratio = 3.81). Those with high values of spherical error had high values of cylindrical error, as well. Mean sphere in WTR, ATR, and oblique astigmatism was 1.93, 1.37, and 0.88 D, respectively (p < 0.001). The highest values of spherical refractive error were observed in WTR astigmatism group. CONCLUSIONS: The prevalence of ATR astigmatism was high in this study. It appears that the decrease in the prevalence of WTR and the increase in the prevalence of ATR astigmatism as a result of aging happened earlier in our study compared with other studies. Astigmatism was found to have a strong correlation with myopia, although individuals with high hyperopia also had high astigmatism. Individuals with high ametropia mostly had WTR astigmatism although the percentage of ATR astigmatism was high in those with low ametropia. PMID- 21623252 TI - Quantified histopathology of the keratoconic cornea. AB - PURPOSE: This study systematically investigated and quantified histopathological changes in a series of keratoconic (Kc) corneas using a physiologically formulated fixative to not further distort the already distorted diseased corneas. METHODS: Twelve surgically removed Kc corneal buttons were immediately preserved and processed for light and transmission electron microscopy using an established corneal protocol. Measurements were taken from the central cone and peripheral regions of the host button. The sample size examined ranged in length from 390 to 2608 MUm centrally and 439 to 2242 MUm peripherally. RESULTS: The average corneal thickness was 437 MUm centrally and 559 MUm peripherally. Epithelial thickness varied centrally from 14 to 92 MUm and peripherally from 30 to 91 MUm. A marked thickening of the epithelial basement membrane was noted in 58% of corneas. Centrally, anterior limiting lamina (ALL) was thinned or lost over 60% of the area examined, whereas peripheral cornea was also affected but to a lesser extent. Histopathologically, posterior cornea remained undisturbed by the disease. Anteriorly in the stroma, an increased number of cells and tissue debris were encountered, and some of these cells were clearly not keratocytes. CONCLUSIONS: It is concluded that Kc pathology, at least initially, has a distinct anterior focus involving the epithelium, ALL, and anterior stroma. The epithelium had lost its cellular uniformity and was compromised by the loss or damage to the ALL. The activity of the hitherto unreported recruited stromal cells may be to break down and remove ALL and anterior stromal lamellae, leading to the overall thinning that accompanies this disease. PMID- 21623253 TI - Importance of rub and rinse in use of multipurpose contact lens solution. AB - PURPOSE: The introduction of contact lens multipurpose disinfection solution (MPDS) that can be used in conjunction with a "no-rub" regimen has simplified lens care requirements. Once adhered to a surface, microorganisms can become less susceptible to disinfection. The aim of the study was to evaluate the effect of various regimen steps on the efficacy of MPDS when used with silicone hydrogel and conventional lenses. METHODS: Commercially available MPDSs containing polyquad or polyhexamethylene biguanide were used in conjunction with two types of silicone hydrogel (lotrafilcon B and galyfilcon A) and one type of conventional soft contact lenses (etafilcon A). Challenge microorganisms included Staphylococcus aureus ATCC 6538, Pseudomonas aeruginosa ATCC 9027, Serratia marcescens ATCC 13880, Fusarium solani ATCC 36031, Candida albicans ATCC 10231, or Acanthamoeba polyphaga Ros. The effect of regimen steps "rub and rinse," "rinse-only," or "no rub and no rinse" on the disinfection efficacy of test MPDSs was examined using the ISO 14729 Regimen Test procedure. RESULTS: Overall, the greatest efficacy of MPDSs was observed when "rub and rinse" was performed before disinfection with each of the microorganisms tested, regardless of lens type. "No rub and no rinse" steps resulted in a greater load of microorganisms remaining on lenses compared with the other regimens (p < 0.05). When "rinse-only" was performed before disinfection, the MPDS containing polyquad performed generally better (p < 0.05) than MPDSs containing polyhexamethylene biguanide against bacteria. Significantly, less microorganisms were recovered from galyfilcon A than from other lenses (p < 0.05) when MPDSs were used with "rinse-only" step. CONCLUSIONS: This study has demonstrated that "rub and rinse" is the most effective regimen and should be recommended in conjunction with all multipurpose lens care solutions and all contact lens types, particularly with silicone hydrogel lenses. PMID- 21623254 TI - Juvenile selective vitamin B12 malabsorption: 50 years after its description-10 years of genetic testing. AB - Fifty years have passed since the description of juvenile selective malabsorption of cobalamin (Cbl). Quality of life improvements have dramatically reduced the incidence of parasite-induced or nutritional Cbl deficiency. Consequently, inherited defects have become a leading cause of Cbl deficiency in children, which is not always expressed as anemia. Unfortunately, the gold standard for clinical diagnosis, the Schilling test, has increasingly become unavailable, and replacement tests are only in their infancy. Genetic testing is complicated by genetic heterogeneity and differential diagnosis. This review documents the history, research, and advances in genetics that have elucidated the causes of juvenile Cbl malabsorption. Genetic research has unearthed many cases in the past decade, mostly in Europe and North America, often among immigrants from the Middle East or North Africa. Lack of suitable clinical testing potentially leaves many patients inadequately diagnosed. The consequences of suboptimal Cbl levels for neurological development are well documented. By raising awareness, we wish to push for fast track development of better clinical tools and suitable genetic testing. Clinical awareness must include attention to ethnicity, a sensitive topic but effective for fast diagnosis. The treatment with monthly parenteral Cbl for life offers a simple and cost-effective solution once proper diagnosis is made. PMID- 21623255 TI - The role of mechanical-electrical interaction in ventricular arrhythmia: evidence from a novel animal model for repaired tetralogy of Fallot. AB - Pulmonary regurgitation and prolonged QRS duration of right bundle branch (RBB) block are common in repaired tetralogy of Fallot (TOF) and increase the risk of sudden death. We sought to establish an animal model to reflect both abnormalities. Twenty-one canines: group I (n = 7) received a surgical right ventricular outflow tract (RVOT) transannular patch plus pulmonary valve destruction; group II (n = 5) received RBB ablation and sham operation; and group III (n = 9) received combined interventions. Serial electrophysiological data were obtained up to 1 y. Procedure mortality was 27.6%. At 1 y, although severe pulmonary regurgitation was documented in most dogs in groups I (71%) and III (100%), progressive RVOT dilatation was noted in group III. RBB block was present in all dogs in groups II and III. However, the increments of QRS duration, QTc, JTc, and QT dispersion progression between 1 mo and 1 y were all greatest in group III. Ventricular arrhythmia events were frequent in group III (median 3.3/mo) but uncommon in groups I and II (median 1/mo). We have created a novel animal model that adequately reflects both the hemodynamic and electrophysiological characteristics of repaired TOF patients and can be applied to examine the risk of ventricular arrhythmias. PMID- 21623256 TI - Indwelling pleural catheter: changing the paradigm of malignant effusion management. PMID- 21623257 TI - Contribution of the TP53, OGG1, CHRNA3, and HLA-DQA1 genes to the risk for lung squamous cell carcinoma. AB - INTRODUCTION: Recent genome-wide association studies (GWASs) have identified polymorphisms in several genes associated with lung cancer risk. Nevertheless, functional polymorphisms in DNA repair and metabolic genes that had been reported as being associated with risk for lung cancer, particularly for lung squamous cell carcinoma (SQC), were not examined in those studies. Therefore, significance of these functional polymorphisms was evaluated in a population, in which polymorphisms in the GWAS genes showed associations with lung SQC risk. METHODS: Polymorphisms in three DNA repair genes, TP53, MDM2, and OGG1, and two metabolic genes, CYP1A1 and GSTM1, were examined for associations with lung SQC risk in a hospital-based case-control study consisting of 377 cases and 325 controls, which had been previously subjected to association studies on GWAS genes, CHRNA3, TERT, and HLA-DQA1. RESULTS: Genotypes for two DNA repair genes, TP53 and OGG1, showed significant associations with SQC risk (p < 0.05), and those for two GWAS genes, CHRNA3 and HLA-DQA1, showed significant associations with SQC risk (P < 0.05) with odds ratios between 1.65 (95% confidence interval = 1.06-2.57 for OGG1) and 2.57 (95% confidence interval = 1.03-6.87 for CHRNA3). Marginally significant associations were also observed for MDM2 and CYP1A1 genes. Interactions among these polymorphisms on SQC risk were not observed. CONCLUSIONS: Association of functional polymorphisms in DNA repair and metabolic genes with lung SQC risk was appreciated. This result indicates the necessity of reevaluation for the significance of functional polymorphisms in DNA repair and metabolic genes on lung cancer risk in other populations subjected to GWASs. PMID- 21623258 TI - Characteristics and outcomes of small cell lung cancer patients diagnosed during two lung cancer computed tomographic screening programs in heavy smokers. AB - INTRODUCTION: Small cell lung cancer (SCLC) is considered an inappropriate screening target due to its short preclinical phase and high rate of relapse despite optimal therapy. However, while intuitively screening for SCLC is inadvisable, in reality, there is a scarcity of data focusing on screen-detected SCLC and whether this intervention leads to diagnosis at an earlier clinical stage or alters outcome. METHODS: We conducted a retrospective review of the baseline characteristics, treatment, and outcome of SCLC patients diagnosed during two large-scale computed tomographic screening studies conducted in heavy smokers. RESULTS: There were 7 of 4782 and 8 of 1520 cases of SCLC identified in the Toronto and Mayo Clinic screening studies, respectively. Complete clinical data were available only for 10 subjects. The median age at diagnosis was 66 years, and 70% were female. The majority were current smokers, with a median pack year history of 50 years. Four cases were detected on enrolment scan, four on annual computed tomography scans, and two on interim scans. Four patients had extensive disease at diagnosis. One of six limited stage patients underwent surgical resection. All 10 patients received first-line chemotherapy. Eight received radiation to at least one site. Eight patients have since died. Median survival was 11.3 months. Two patients remain disease free at 2 and 9 years, respectively. CONCLUSION: This study suggests that computed tomography screening is ineffective for SCLC. Efforts to reduce mortality of SCLC should instead focus on prevention through tobacco reduction programs, as well as the development of improved treatment options. PMID- 21623259 TI - Vertebral collapse caused by bone metastasis. PMID- 21623260 TI - Treatment of cardiac angiosarcoma with radiation and docetaxel: a case report with partial response and prolonged stable disease. PMID- 21623262 TI - Response to the article "pulmonary resection for metastatic gastric cancer" by kemp et Al. PMID- 21623264 TI - The diverse diversity. PMID- 21623265 TI - Activity of crizotinib (PF02341066), a dual mesenchymal-epithelial transition (MET) and anaplastic lymphoma kinase (ALK) inhibitor, in a non-small cell lung cancer patient with de novo MET amplification. AB - Crizotinib is a dual MET and ALK inhibitor. Currently, clinical development of crizotinib is focused primarily on ALK rearranged non-small cell lung cancer (NSCLC). Here we report an NSCLC patient with de novo MET amplification but no ALK rearrangement who achieved a rapid and durable response to crizotinib indicating is also a bona fide MET inhibitor. PMID- 21623266 TI - EGFR gene alterations in a Norwegian cohort of lung cancer patients selected for surgery. AB - INTRODUCTION: Lung cancer is the leading cause of cancer-related deaths worldwide. New therapies targeting the epidermal growth factor receptor (EGFR) tyrosine kinase are promising and show high response rates in the subset of patients with activating mutations in EGFR. The frequency of these mutations is largely unknown in unselected Caucasian patients. METHODS: Mutation analysis of EGFR exons 18-21 was performed on 240 lung cancer samples using the TheraScreen EGFR mutation kit and denaturing high-performance liquid chromatography in addition to sequencing. RESULTS: In a cohort of 240 Norwegian lung cancer patients selected for surgery, we identified 18 tumors with EGFR-activating mutations (7.5%, 14 women and 4 men), of which 14 were adenocarcinomas, 2 squamous cell carcinomas, and 2 bronchoalveolar carcinomas. Five of the mutations were found in patients with more than 20 pack-years of smoking history. CONCLUSION: The frequency of EGFR mutations is lower in our cohort than among Asian lung cancer patients and present in both men and women and smokers and never-smokers. However, the frequency is significantly higher among women and never-smokers and among patients with adenocarcinomas. PMID- 21623267 TI - Phase I study of sunitinib and erlotinib in advanced nonsquamous non-small cell lung cancer. AB - INTRODUCTION: Erlotinib has prolonged survival in unselected patients with advanced non-small cell lung cancer, whereas sunitinib has yielded promising rates of disease control in previously treated patients. We conducted a dose escalation study of this combination to determine the maximum tolerated dose of sunitinib in combination with a fixed dose of erlotinib and to evaluate the toxicities of this combination. METHODS: Patients with advanced nonsquamous non small cell lung cancer were treated at two dose levels: sunitinib at either 25 mg or 37.5 mg, with erlotinib 150 mg. Both drugs were given once daily, continuously. RESULTS: Eleven patients enrolled from November 2007 to October 2009. No dose-limiting toxicities occurred. Grade 3/4 adverse events at least possibly related to treatment were seen in seven patients (64%). Six patients (54%) required dose modifications, and three (27%) discontinued study treatment due to toxicity. Rates of grade 3 diarrhea and mucositis exceeded those seen with single-agent erlotinib or sunitinib. One patient (9%) attained a partial response lasting 16.3 months. CONCLUSIONS: Although no dose-limiting toxicities occurred, it is difficult to recommend erlotinib 150 mg and sunitinib 37.5 mg daily as the phase II dose for this combination due to the high rate of adverse events. Because of the overlapping toxicity profile of each agent, this combination was poorly tolerated in our population. PMID- 21623268 TI - Palmar bullous blistering induced by erlotinib. PMID- 21623269 TI - Complete radiological response of metastatic anaplastic lymphoma kinase-positive signet ring lung adenocarcinoma to systemic chemotherapy. PMID- 21623270 TI - Isolated epithelioid trophoblastic tumor mimicking non-small cell lung cancer. PMID- 21623271 TI - Paraneoplastic opsoclonus-myoclonus syndrome as a new and single manifestation of relapsing disease in a patient with small cell lung cancer. PMID- 21623272 TI - Angiosarcoma arising from right atrium: remarkable response to concurrent chemoradiotherapy with carboplatin and paclitaxel. PMID- 21623273 TI - Clinical significance of serum vascular endothelial growth factor in malignant pleural mesothelioma. PMID- 21623274 TI - Chemotherapy dose intensity and survival in non-small cell lung cancer. PMID- 21623275 TI - How do we do it?: another optimal methodology for endobronchial ultrasound sample handling. PMID- 21623276 TI - Split-course radiotherapy for poor-risk stage III lung cancer: striking the right balance or history regurgitated? PMID- 21623277 TI - Biostatistics primer: what a clinician ought to know: hazard ratios. AB - Hazard ratios (HRs) are used commonly to report results from randomized clinical trials in oncology. However, they remain one of the most perplexing concepts for clinicians. A good understanding of HRs is needed to effectively interpret the medical literature to make important treatment decisions. This article provides clear guidelines to clinicians about how to appropriately interpret HRs. While this article focuses on the commonly used methods, the authors acknowledge that other statistical methods exist for analyzing survival data. PMID- 21623278 TI - The tumor necrosis factor-related apoptosis-inducing ligand and lung cancer: still following the right TRAIL? AB - Tumor necrosis factor-related apoptosis-inducing ligand is a type II membrane bound protein whose C-terminal extracellular domain shows clear homology to other tumor necrosis factor family members. It is constitutively expressed on macrophages, T cells, natural killer cells, and dendritic cells and selectively kills transformed cells leaving most of the normal cells alone. This selectivity has led to great interest in it use as a therapeutic agent for the treatment of malignancy. In this review, this critical pathway is described, highlighting its mechanistic manipulation for therapeutic benefit and the recent phase I and II trials in lung cancer that have been performed or are currently ongoing are also discussed. PMID- 21623280 TI - Successful erlotinib rechallenge after erlotinib-induced interstitial lung disease. PMID- 21623279 TI - Phase II trial of dasatinib for patients with acquired resistance to treatment with the epidermal growth factor receptor tyrosine kinase inhibitors erlotinib or gefitinib. AB - INTRODUCTION: Dual inhibition of SRC- and EGFR-dependent pathways may overcome acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) for patients with lung adenocarcinoma with EGFR mutations. The SRC inhibitor dasatinib demonstrates antitumor activity in gefitinib resistant cells lines and xenografts. Dasatinib is tolerable for patients with advanced non-small cell lung cancer, and in combination with erlotinib. METHODS: We conducted this phase II study of dasatinib 70 mg twice daily in patients with EGFR-mutant lung adenocarcinoma and acquired resistance to EGFR-TKIs. After a protocol amendment based on evolving data about both drugs, patients received dasatinib at a dose of 100 mg daily with continued erlotinib after developing acquired resistance. Enrolled patients either harbored an activating mutation in EGFR or experienced clinical benefit with single-agent erlotinib or gefitinib, followed by RECIST documented progression while being treated with an EGFR-TKI. RESULTS: Twenty-one patients were enrolled, 9 under the original trial design and 12 after the protocol amendments. We observed no complete or partial responses (0% observed rate, 95% confidence interval: 0-18%). The median time to progression was 0.5 months (range, 0.2-1.8 months) in patients treated with dasatinib and 0.9 months (range, 0.4-5 months) for patients treated with dasatinib and erlotinib in combination. Pleural effusions and dyspnea were frequent toxicities. CONCLUSIONS: Dasatinib has no activity in patients with EGFR mutant lung adenocarcinoma with acquired resistance to erlotinib and gefitinib. PMID- 21623281 TI - Do complex mutations of the epidermal growth factor receptor gene reflect intratumoral heterogeneity? PMID- 21623282 TI - Critical care for patients with lung cancer. PMID- 21623283 TI - Evaluation of ALK gene status in primary lung adenocarcinoma and matched metastases. PMID- 21623286 TI - The faces of fatigue. PMID- 21623287 TI - Sport-specific aquatic rehabilitation. PMID- 21623288 TI - Making prudent recommendations for return-to-play in adult athletes with cardiac conditions. AB - Clinicians who treat millions of adult athletes throughout the world may be faced with participation or return-to-play decisions in individuals with known or suspected cardiac conditions. Here we review existing published participation guidelines and analyze emerging data from ongoing registries and population-based studies pertaining to return-to-play decisions for cardiac conditions specifically affecting adult athletes. Considerations related to return-to-play decisions will vary according to age of the athlete, with inherited disorders being the main consideration in younger adult athletes aged 18 to 40 yr, and coronary artery disease being the main consideration in older adult athletes aged 40 yr and older. Although this arbitrary division is based on the epidemiology of underlying heart disease in these populations, the essential return-to-play decision process for both age groups is quite similar. Among the most widely used guidelines to make return-to-play decisions in this group of athletes are the 36th Bethesda Conference Eligibility Recommendations for Competitive Athletes with Cardiovascular Abnormalities. These have long been considered the "gold standard" for determining return-to-play decisions in young athletes in the United States. Other guidelines are available for unique purposes, including The European Society of Cardiology guidelines, and the American Heart Association published recommendations regarding participation of young patients (younger than 40 yr) with genetic cardiovascular diseases in recreational sports. The latter are consistent with the 36th Bethesda guidelines and cover common genetically based diseases such as inherited cardiomyopathies, channelopathy, and connective tissue disorders like Marfan's syndrome. The consensus on masters athletes (older than 40 yr) provides return-to-play decisions for a wide variety of conditioned states, from elite older athletes to walk-up athletes. For any adult athlete with a cardiac condition, return-to-play decisions following use of medications, ablation procedures, device implantation, corrective surgery, or coronary intervention depend on whether the procedure has sufficiently altered the risk for sudden cardiac events, and whether there is a potential for unfavorable interaction with cardiac performance. PMID- 21623289 TI - Infectious disease in athletes. AB - While orthopedic injuries most commonly are associated with sports, infectious diseases cause significant morbidity in athletes. Exercise improves immunity at moderate intensity but impairs immune function at extremes of duration and intensity. Respiratory infections are the most common, but skin, blood borne, sexually transmitted, and even cardiac infections occur. Infectious disease outbreaks are a constant concern. Treatment of such infections resembles those used in the general population. Return to play issues and prevention of infection are especially important in athletes. PMID- 21623290 TI - Practical recommendations and perspectives on cardiac screening for healthy pediatric athletes. AB - Primary care providers often are asked to perform the preparticipation physical exam and cardiac screening of pediatric athletes. It can be challenging to evaluate which athletes may require further cardiac evaluation and specialist referral based on a focused history and physical. There is considerable controversy surrounding mass electrocardiogram (ECG) screening of athletes in the United States. The ECG is one of several diagnostic tests used by pediatric cardiologists in the evaluation of underlying heart disease in young athletes. This article reviews cardiac conditions associated with sudden death, discusses pertinent findings on history and physical, and provides clinically relevant information for practitioners who are deciding which patients to refer for detailed cardiac evaluation. PMID- 21623291 TI - Ultrasound-guided percutaneous neuroplasty of the lateral femoral cutaneous nerve for the treatment of meralgia paresthetica: a case report and description of a new ultrasound-guided technique. AB - The lateral femoral cutaneous nerve (LFCN) can be visualized with ultrasound imaging using a high frequency linear transducer. The entrapment of the LFCN, often near the lateral aspect of the inguinal ligament, is accepted as an etiology of meralgia paresthetica (MP). This case report describes an ultrasound guided, percutaneous technique that utilizes injected fluid to facilitate blunt dissection (or hydrodissection) to perform an external neuroplasty of the LFCN. This procedure resulted in immediate, long-term relief of pain associated with severe, chronic MP. This procedure may potentially represent an alternate treatment for patients with contraindications or partial contraindications to surgical neurolysis, nerve transection (usually described as being performed under general anesthesia), or corticosteroid injection, or in patients not responding to conservative treatment measures. PMID- 21623292 TI - Celiac disease and the athlete. AB - With the diagnosis of celiac disease rising in the past decade and with increased public awareness, team physicians are faced with both managing and diagnosing athletes with celiac disease. Sports medicine physicians need to recognize that celiac disease can present with a number of different symptoms and, therefore, should consider celiac disease as part of their differential in evaluating athletes with prolonged unexplained illnesses. Sports medicine physicians must be familiar with the appropriate laboratory tests and diagnostic procedures used to establish the diagnosis of celiac disease. A multidisciplinary approach in helping the newly diagnosed athlete with celiac disease is important to the successful treatment of the disease. Athletes with celiac disease often have problems with iron absorption (leading to anemia) and/or vitamin D and calcium absorption (leading to osteoporosis and poor bone health). Even athletes with known and long-standing celiac disease need additional care and supervision in ensuring there is no disruption in their gluten-free diet, which can lead to a flare-up of symptoms or a decrease in performance. PMID- 21623293 TI - Evaluation and treatment of GERD and upper GI complaints in athletes. AB - Athletes are susceptible to upper gastrointestinal complaints just like the general population. The most common etiologies are gastroesophageal reflux disease (GERD) and functional heartburn. If the signs and symptoms are compatible with GERD and the clinician has considered more serious pathology unlikely, a therapeutic trial with a proton pump inhibitor (PPI) can be initiated without further evaluation. The treatment for pure exertional GERD is similar but is guided by expert opinion only. Surgery has a limited role, but new techniques are evolving that may change the risk-to-benefit ratio. Chronic PPI therapy generally is safe, but there is a small risk of osteoporosis with concomitant fracture. There is no evidence for routine endoscopic screening for Barrett's esophagus or esophageal adenocarcinoma. For those who do not respond to treatment, the most likely diagnosis is functional heartburn. This is a diagnosis of exclusion, and referral to gastroenterology is warranted for diagnostic testing. PMID- 21623294 TI - Lumbar plexus nerve entrapment syndromes as a cause of groin pain in athletes. AB - In athletes, groin pain is not uncommon and can be severe and activity-limiting. Nerve entrapment syndromes of the lumbar plexus are a rare but important etiology that should be considered when evaluating athletes. Diagnosis can be made based on patterns of pain and hypoesthesia following the sensory distribution of the involved nerve and by pain relief with nerve block. Conservative therapies, including nerve blocks, neurodestructive procedures, and medications, may provide long-term pain relief. If nonsurgical therapies fail, referral should be made for surgical exploration and neurectomy. PMID- 21623295 TI - ACSM Clinician Profile. PMID- 21623296 TI - Thromboembolic disorders: guidance for return-to-play. AB - Venous thromboembolism (VTE) is a major cause of morbidity and mortality. Treatment for VTE in athletes is similar to nonathletes. Early treatment of deep venous thrombosis (DVT) with bed rest and anticoagulation has given way to anticoagulation with early mobilization. Thrombolysis, preferably catheter directed thrombolysis (CDT), may be used in select patients with upper extremity DVT (UEDVT). Surgical procedures should be reserved for those athletes with UEDVT who fail initial therapy. Compression devices are advocated for the treatment of postthrombotic symptoms (PTS) in lower extremity DVT (LEDVT) and UEDVT. Athletes with DVT should be encouraged to start a gradual return to activities of daily living (ADL) the day they begin anticoagulation therapy. A structured return-to training program with progressive increase in intensity can begin shortly after ADL mastery, provided the athlete is monitored carefully for recurrence of VTE. Athletes should not engage in contact or collision sports until anticoagulation therapy is complete. PMID- 21623297 TI - Overcaffeination: low potassium and other perils. PMID- 21623298 TI - Genetics and soft-tissue injuries in sport: clinical commentary. PMID- 21623299 TI - Traumatic retinal tear in a basketball player. PMID- 21623300 TI - Sideline and event management in golf. AB - Golf has great popularity as a spectator sport, as well as a participation sport. Providing coverage for golf events can range from mass event coverage for large professional tournaments to provision of injury care at local golf events. This article provides a brief introduction to the game of golf for those unfamiliar with its play, an overview of the types of injuries seen, and consideration in providing care for a variety of golf competitions. PMID- 21623301 TI - SCUBA medicine: a first-responder's guide to diving injuries. AB - Self-contained underwater breathing apparatus (SCUBA) diving is an ever-growing sport, and despite a myriad of technological advances to improve safety, it remains dangerous. Providers of medical care for SCUBA divers must have an understanding of diving physiology and potential medical problems that can occur. SCUBA diving also can take participants to remote areas, so being properly prepared for potential emergencies can make a significant difference. The following is a review of diving physiology and the medical problems that can occur in SCUBA divers, along with some suggestions as to how to prepare for a SCUBA excursion. PMID- 21623302 TI - Mixed martial arts: injury patterns and issues for the ringside physician. AB - Mixed martial arts (MMA) is a fast-growing combat sport. There are limited studies investigating MMA's injury incidence. However, from the data available, closed head injuries and lacerations commonly are experienced by competitors. Sanctioned events require the presence of ringside physicians, who should be well versed in handling these types of injuries. Additionally, sports-medicine providers should have an appropriately equipped medical bag, and before providing coverage, they need to coordinate with emergency medical personnel in order to more efficiently deliver care if urgent situations arise. More research is needed on injury incidence and prevention in MAA. PMID- 21623303 TI - Reducing the risk of falls among older adults: the Fallproof Balance and Mobility Program. AB - Fall-related deaths among the older adult segment of the population constitute a growing public health concern that is largely preventable. A growing body of research has identified a number of effective intervention strategies that can lower the incidence of falls and/or risk factors that contribute to heightened fall risk. One particularly effective intervention strategy that has been identified is exercise, whether individually prescribed or conducted in group based settings. The Fallproof Balance and Mobility Program was developed in response to the need for effective community-based programs that target the important intrinsic risk factors (e.g., impaired balance and gait, muscle weakness) associated with increased fall risk. This theory-driven program adopts a multidimensional approach to balance and mobility that has proven to be effective in reducing fall risk among older adults identified as moderate-to-high risk for falls. The fidelity of the program also has been maintained by implementing an instructor certification program and standardizing program content and delivery. PMID- 21623304 TI - ACSM Clinician Profile. AB - The American College of Sports Medicine (ACSM) is proud to feature Elizabeth A. Joy, MD, MPH, FACSM, for this issue's Clinician Profile. Dr. Joy is a family medicine and sports medicine physician at the University of Utah,where she serves as one of the team physicians and directs the Primary Care Sports Medicine Fellowship program. She completed a Family Medicine Residency and Primary Care Sports Medicine Fellowship at Hennepin County Medical Center in Minneapolis, MN. She completed her master's degree in public health at the University of Utah. Dr. Joy has served on ACSM's Board of Trustees and is currently the Vice-President for Medicine, serves on ACSM's 'Exercise is Medicine' Task Force, and is an associate editor for Current Sports Medicine Reports (CSMR). Her research interests include physical activity assessment and promotion, practice-based research in primary care, the Female Athlete Triad, exercise during pregnancy, and injury prevention. PMID- 21623306 TI - On-mountain coverage of competitive skiing and snowboarding events. AB - Skiing and snowboarding are popular sports worldwide both for competitive and recreational participants. Medical coverage of the competitive events can be challenging because of the wide variety of competition styles and venues. Skiing and snowboarding have similar overall rates of injury, with lower extremity injuries more prevalent in skiing and upper extremity injuries in snowboarding. Providers of medical coverage for these events usually have to be skilled in skiing and/or snowboarding to reach injured athletes or sometimes even to get to the event venue. Care of even routine injuries can be challenging because of environmental conditions and terrain. Catastrophic injuries fortunately are rare and are most commonly related to head trauma. Spine injuries and thoracoabdominal trauma also are seen. Because remote mountainous locations are common, advance planning for the treatment of all injuries, particularly serious or catastrophic, is paramount. Common nonorthopedic conditions include altitude sickness, cold related conditions, and other travel-related illnesses, such as jet lag and food borne illness. PMID- 21623308 TI - Firefighter fitness: improving performance and preventing injuries and fatalities. AB - Firefighting is dangerous work. Each year, approximately 80,000 firefighters are injured and about 100 firefighters lose their lives in the line of duty. Firefighters face multiple dangers in the course of their work; they encounter toxic fumes, dangerous products of combustion, high radiant heat loads, and a chaotic work environment. Despite the myriad dangers, the leading cause of line of-duty death among firefighters is sudden cardiac event, accounting for approximately 45% of duty deaths. Firefighting requires high levels of aerobic fitness, anaerobic capacity, and muscular strength and endurance; however, data suggest that many firefighters do not possess high aerobic or anaerobic capacity. Furthermore, many firefighters are overweight and have one or more modifiable risk factors for cardiovascular disease. The safety of the public and the health and safety of firefighters would be enhanced if firefighters followed well designed fitness programs to improve overall health and fitness. PMID- 21623307 TI - When to initiate integrative neuromuscular training to reduce sports-related injuries and enhance health in youth? AB - Regular participation in organized youth sports does not ensure adequate exposure to skill- and health-related fitness activities, and sport training without preparatory conditioning does not appear to reduce risk of injury in young athletes. Current trends indicate that widespread participation in organized youth sports is occurring at a younger age, especially in girls. Current public health recommendations developed to promote muscle strengthening and bone building activities for youth aged 6 yr and older, along with increased involvement in competitive sport activities at younger ages, has increased interest and concern from parents, clinicians, coaches, and teachers regarding the optimal age to encourage and integrate more specialized physical training into youth development programs. This review synthesizes the latest literature and expert opinion regarding when to initiate neuromuscular conditioning in youth and presents a how-to integrative training conceptual model that could maximize the potential health-related benefits for children by reducing sports-related injury risk and encouraging lifelong, regular physical activity. PMID- 21623309 TI - Knoevenagel reaction in [MMIm][MSO4]: synthesis of coumarins. AB - The ionic liquid 1,3-dimethylimidazolium methyl sulfate, [MMIm][MSO4], together with a small amount of water (the amount taken up by the ionic liquid upon exposure to air), acts efficiently as both solvent and catalyst of the Knoevenagel condensation reactions of malononitrile with 4-substituted benzaldehydes, without the need for any other solvent or promoter, affording yields of 92%-99% within 2-7 min at room temperature. When L-proline is used as an additional promoter to obtain coumarins from o-hydroxybenzaldehydes, the reaction also proceeds in high yields. Work-up is very simple and the ionic liquid can be reused several times. Some of the coumarins obtained are described for the first time. PMID- 21623310 TI - In vitro and in vivo antitumor activity of Scutellaria barbate extract on murine liver cancer. AB - In the present study, we investigated the in vitro and in vivo antitumor effects of crude extract of Scutellaria Barbate (CE-SB) on mouse hepatoma H22 cells. The MTT assay was used to determine the growth inhibition of H22 cells in vitro. The in vivo therapeutic effects of CE-SB were determined using H22 tumor bearing mice. Besides, the body weight, tumor weight, thymus index and spleen index of H22 bearing mice were also measured. The tumor inhibitory rate (IR) was calculated according to the mean weight of tumor (MWT). The phagocytotic function of macrophages was examined by observing peritoneal macrophages phagocytize chicken RBC. The results showed that CE-SB could inhibit the growth of hepatoma H22 Cells in vitro and in vivo. Furthermore, CE-SB could improve immune function of H22 tumor bearing mice. Together these results indicate that CE-SB has antitumor activity and seems to be safe and effective for the use of anti-tumor therapy. PMID- 21623311 TI - Two new acridone alkaloids from Glycosmis macrantha. AB - Extraction and chromatographic separation of the extracts of dried stem barks of Glycosmis macrantha lead to isolation of two new acridone alkaloids, macranthanine and 7-hydroxynoracronycine, and a known acridone, atalaphyllidine. The structures of these alkaloids were determined by detailed spectral analysis and also by comparison with reported data. PMID- 21623312 TI - Targeting oncogenic protein-protein interactions by diversity oriented synthesis and combinatorial chemistry approaches. AB - We are currently witnessing a decline in the development of efficient new anticancer drugs, despite the salient efforts made on all fronts of cancer drug discovery. This trend presumably relates to the substantial heterogeneity and the inherent biological complexity of cancer, which hinder drug development success. Protein-protein interactions (PPIs) are key players in numerous cellular processes and aberrant interruption of this complex network provides a basis for various disease states, including cancer. Thus, it is now believed that cancer drug discovery, in addition to the design of single-targeted bioactive compounds, should also incorporate diversity-oriented synthesis (DOS) and other combinatorial strategies in order to exploit the ability of multi-functional scaffolds to modulate multiple protein-protein interactions (biological hubs). Throughout the review, we highlight the chemistry driven approaches to access diversity space for the discovery of small molecules that disrupt oncogenic PPIs, namely the p53-Mdm2, Bcl-2/Bcl-xL-BH3, Myc-Max, and p53-Mdmx/Mdm2 interactions. PMID- 21623313 TI - Microwave-assisted extraction of polyphenols from Camellia oleifera fruit hull. AB - The abundant fruit hulls of tea-oil tree (Camellia oleifera) are still underutilized and wastefully discaded to pollute the environment. In order to solve this problem and better utilize the fruit hulls of C. oleifera, a microwave assisted extraction system was used to extract their polyphenols using water as the extraction solvent. A central composite design (CCD) was used to monitor the effects of three extraction processing parameters--liquid:solid ratio (mL/g), extraction time (min) and extraction temperature ( degrees C)--on the polyphenol yield (%). The results showed that the optimal conditions were liquid:solid ratio of 15.33:1 (mL/g), extraction time of 35 min and extraction temperature of 76 degrees C. Validation tests indicated that under the optimized conditions the actual yield of polyphenols was 15.05 +/- 0.04% with RSD = 0.21% (n = 5), which was in good agreement with the predicted yield. Phenolic compounds in the extracts were analysed by HPLC, and gallic acid was found to be the predominant constituent. The total flavonoid content in the extracts was determined and high total flavonoid content was revealed (140.06 mg/g dry material). PMID- 21623314 TI - Phytochemical analysis and antimicrobial activities of methanolic extracts of leaf, stem and root from different varieties of Labisa pumila Benth. AB - A local herb, Kacip Fatimah, is famous amongst Malay women for its uses in parturition; however, its phytochemical contents have not been fully documented. Therefore, a study was performed to evaluate the phenolics, flavonoids, and total saponin contents, and antibacterial and antifungal properties of the leaf, stem and root of three varieties of Labisia pumila Benth. Total saponins were found to be higher in the leaves of all three varieties, compared to the roots and stems. Leaves of var. pumila exhibited significantly higher total saponin content than var. alata and lanceolata, with values of 56.4, 43.6 and 42.3 mg diosgenin equivalent/g dry weight, respectively. HPLC analyses of phenolics and flavonoids in all three varieties revealed the presence of gallic acid, caffeic acid, rutin, and myricetin in all plant parts. Higher levels of flavonoids (rutin, quercitin, kaempferol) were observed in var. pumila compared with alata and lanceolata, whereas higher accumulation of phenolics (gallic acid, pyrogallol) was recorded in var. alata, followed by pumila and lanceolata. Antibacterial activities of leaf, stem and root extracts of all varieties determined against both Gram positive (Micrococcus luteus, Bacillus subtilis B145, Bacillus cereus B43, Staphylococcus aureus S1431) and Gram negative (Enterobacter aerogenes, Klebsiella pneumonia K36, Escherichia coli E256, Pseudomonas aeruginosa PI96) pathogens showed that crude methanolic extracts are active against these bacteria at low concentrations, albeit with lower antibacterial activity compared to kanamycin used as the control. Antifungal activity of methanolic extracts of all plant parts against Fusarium sp., Candida sp. and Mucor using the agar diffusion disc exhibited moderate to appreciable antifungal activities compared to streptomycin used as positive control. PMID- 21623315 TI - Bismuth modified carbon-based electrodes for the determination of selected neonicotinoid insecticides. AB - Two types of bismuth modified electrodes, a bismuth-film modified glassy carbon (BiF-GCE) and a bismuth bulk modified carbon paste, were applied for the determination of selected nitroguanidine neonicotinoid insecticides. The method based on an ex situ prepared BiF-GCE operated in the differential pulse voltammetric (DPV) mode was applied to determine clothianidin in the concentration range from 2.5 to 23 MUg cm-3 with a relative standard deviation (RSD) not exceeding 1.5%. The tricresyl phosphate-based carbon paste electrodes (TCP-CPEs), bulk modified with 5 and 20 w/w% of bismuth, showed a different analytical performance in the determination of imidacloprid, regarding the peak shape, potential window, and noise level. The TCP-CPE with 5% Bi was advantageous, and the developed DPV method based on it allowed the determination in the concentration range from 1.7 to 60 MUg cm-3 with an RSD of 2.4%. To get a deeper insight into the morphology of the bismuth-based sensor surfaces, scanning electron microscopic measurements were performed of both the surface film and the bulk modified electrodes. PMID- 21623316 TI - A novel thiophene-fused polycyclic aromatic with a tetracene core: synthesis, characterization, optical and electrochemical properties. AB - FeCl3-mediated oxidative cyclization was successfully used to construct an extended thiophene-pendant pyrene skeleton and synthesize a novel thiophene-fused polycyclic aromatic (THTP-C) with a tetracene core. The identity of the compound was confirmed by 1H-NMR, 13C-NMR, MS, and elemental analysis. Meanwhile, a single crystal of THTP-C was obtained and analyzed by X-ray single-crystal diffraction. THTP-C has a "saddle" shaped pi-conjugated 1-D supramolecular structure, and favors highly ordered self-assembly by pi-pi interactions as evidenced by its concentration-dependent 1H-NMR spectra in solution. The optical properties of THTP-C were investigated by ultraviolet-visible (UV-Vis) and photoluminescence (PL) spectroscopy and its electrochemical properties were investigated by cyclic voltammetry (CV). The relatively large band gap (2.86 eV), low E(HOMO) level ( 5.64 eV) and intermolecular pi-pi interactions imply that THTP-C has a high stability against photo-degradation and oxidation, and may be a promising candidate for stable hole-transporting materials. PMID- 21623317 TI - Pharmaceutical composition of hydrochlorothiazide:beta-cyclo-dextrin: preparation by three different methods, physico-chemical characterization and in vivo diuretic activity evaluation. AB - Hydrochlorothiazide is a common diuretic antihypertensive drug of the thiazide family. Its poor aqueous solubility is one of the reasons for its limited bioavailability after oral administration. This work aimed at the development of a hydrochlorothiazide:beta-cyclodextrin (HTZ:beta-CD) pharmaceutical composition in order to improve water solubility and bioavailability of the drug. The HTZ:beta-CD complexes were prepared by three different methods: spray-drying, freeze-drying and fluid bed. Complexes were characterized by thermal analysis, Fourier transform-infrared (FTIR) spectroscopy, powder X-ray diffractometry, NMR (2D-ROESY), scanning electron microscopy (SEM), particle analysis and intrinsic dissolution. The findings reveal that three binary systems prepared presented better solubility results in comparison with free HTZ. Increased diuretic effect was observed to HTZ:beta-CD obtained by fluid bed in comparison to free drug in rats. Results taken together suggest that pharmacological effect of HTZ in complex was increased by solubility improvement promoted by cyclodextrin. PMID- 21623318 TI - Catalytic synthesis of alpha-oxoketene S,S-acetals in a wet ionic liquid [Bmim]Cl/H2O homogeneous system. AB - A clean, practical and environmentally friendly synthesis in a homogeneous system has been developed for alpha-oxoketene S,S-acetals. A mixture of [Bmim]Cl and water was used as medium. The best economical and practical molar ratio of [Bmim]Cl to substrate was 4 to 1. With various types of 1,3-dicarbonyl compounds as substrate, the corresponding alpha-oxoketene S,S-acetals have been prepared under mild reaction conditions with yields of 53-74% after purification with silica gel column. [Bmim]Cl/water can be recycled several times. PMID- 21623319 TI - Guidance of magnetic resonance imaging and placement of skin-marker localization devices. AB - AIM: A variety of magnetic resonance imaging (MRI)-compatible skin-marker localization devices are available on the market. MRI protocols call for the liberal use of the skin markers over the specific site of symptoms or over any palpable mass. This study investigates the usefulness of patient-assisted placement of 1 000-mg fish oil capsules as skin markers over the area of maximum localized pain, signs, or symptoms and correlates this placement with any potential underlying neuropathology or potential pain generator. METHODS: One hundred symptomatic patients undergoing MRI were assessed for focal or localized signs or symptoms. Under the direction of a physician and with guidance from the patient, the MRI technician placed a 1 000-mg fish-oil capsule over the area of maximum pain or signs and symptoms. Patients with poorly localized, diffuse symptoms or an area of maximal signs and symptoms outside the field of view of the MRI were not included in this study. All MRI exams were reviewed by clinical physicians and radiologists or neuroimaging physicians. RESULTS: In all 100 cases, the images show clearly visible MRI-compatible skin-surface markers that correlate with potential underlying neuropathology. CONCLUSION: Our results show that 1 000-mg fish-oil capsules can be used as MRI localization devices as a cost effective alternative to more expensive commercially available devices. PMID- 21623320 TI - Prevention of post-operative infection in spine surgery by wound irrigation with a solution of povidone-iodine and hydrogen peroxide. AB - Starting from January 2008, we systematically irrigated the surgical wounds of patients undergoing spine surgery with a solution of povidone-iodine and hydrogen peroxide. We prospectively recorded the clinical data of patients who underwent spine surgery during 2008 and we compared the results with retrospectively reviewed clinical records of patients operated during 2007. Patients were analyzed for preoperative risk factors, type of surgical procedure, onset of infection, presentation, treatment and outcome. We performed 490 spine surgeries during 2008 and 460 during 2007. We recorded four postoperative infections in 2007 compared to none in 2008. We consider this solution of povidone-iodine plus hydrogen peroxide effective to further reduce the rate of post-operative infection in spine surgery. PMID- 21623321 TI - Posterior interosseous nerve palsy by synovial cyst of proximal radioulnar joint: our experience after 5 years. AB - AIM: The posterior interosseous nerve palsy is a neuropathy of radial nerve interesting its deep motor branch. The neuropathy can appear with a hollow in the proximal half of the forearm without significant swelling, a complete loss of extension of the fingers with radial deviation of the wrist during extension. In some cases, PIN compression may simulate tendon rupture in rheumatologic diseases, because the pain and the paralysis occur suddenly, so often can be difficult to make a diagnosis. The palsy is caused by compression of the posterior interosseous nerve from soft tissue tumours or tumour-like masses: ganglions, lipomas, rheumatoid synovitis, synovial chondromatosis, fibromas, neurofibromas, bursitis, synovial cysts of the elbow and radioulnar proximal joints. The aim of our research was to individuate the better treatment for the posterior interosseous nerve palsy. METHODS: From 2002 to 2007 we examined 8 patients: 2 female and 6 male. Median age was 43 years. The diagnosis was made by clinical examination, ultrasound, nerve conduction studies and magnetic resonance imaging (MRI). Patients underwent to decompressing posterior interosseous nerve surgery. RESULTS: After the surgical exploration in 8 cases a globular mass of around 2.5 cm to 4.5 cm diameter was discovered. At the histological examination, a synovial cyst of the elbow joint was found in 7 out of 8 patients and an hemangioma tumor in the one remaining patient. 12 months was the median time for a complete recovery after the operation, confirmed by EMG. CONCLUSION: The surgical treatment offers a complete resolution in all cases. PMID- 21623322 TI - Intracranial cerebral revascularization. AB - Stroke is a heterogeneous disorder. A small number are due to hemodynamic insufficiency. In these cases, blood supply is sufficient at rest but unable to meet increased demand. This is demonstrated by loss of cerebrovascular reactivity (CVR) or increased oxygen extraction fraction (OEF). This may be improved by a superficial temporal artery to middle cerebral artery bypass. Whilst too small to completely replace cerebral blood supply, this is sufficient to augment existing blood flow to meet any increases in demand. Practically it has been difficult to demonstrate any reduction in stroke rate with intracranial revascularization. Only one of three randomized studies has shown a benefit. Two problems have been how hemodynamic insufficiency and symptomatic patients have been defined. Hemodynamic insufficiency can only be reliably demonstrated by changes in OEF and CVR. Symptomatic has traditionally described a patient who was had one or more strokes. However, in most cases these occur at the time of occlusion following which the patient remains stable. These patients are not necessarily at a high risk of future stroke as collaterals develop over time to compensate. We prefer to reserve the term symptomatic for those who have ongoing fluctuating symptoms after demonstration of carotid occlusion. This much smaller subset may still benefit from surgery in our view. Therefore although bypass surgery has relatively low morbidity, and high graft patency, extremely careful patient selection is essential for it to benefit patients, and unless there are both severe hemodynamic insufficiency and ongoing symptoms medical management is preferable. PMID- 21623323 TI - Cerebral revascularization: extracranial-intracranial bypass. AB - Extracranial-intracranial (EC-IC) bypass remains an important revascularization technique for management of complex cerebrovascular disease. Despite evolving endovascular techniques, the role of bypass for the purpose of flow replacement prior to planned vessel sacrifice remains relevant for treatment of complex and fusiform aneurysms. The role of bypass for purposes of flow augmentation in the setting of cerebral ischemia is limited based on current data, but remains an important option for selected cases of athero-occlusive disease, in addition to a primary treatment for symptomatic moyamoya disease. An objective flow-based approach to EC-IC bypass can enhance decision-making in preoperative patient selection, intraoperative graft assessment, and postoperative follow-up. PMID- 21623324 TI - Towards sutureless non-occlusive cerebral revascularization. AB - Sutureless vascular anastomoses have been the subject of extensive research for decades. In neurosurgery the need for a safe and fast technique is high, because temporary occlusion of cerebral arteries may rapidly lead to brain ischemia. Conventional sutured anastomoses have always been the golden standard. Limited working space and difficult suturing techniques were reasons to find alternatives. Many artificial devices to create anastomoses have been engineered over the years like tissue sealants, clips and automated suturing sets with variable success. For all previous options, temporary occlusion of the recipient artery was necessary. The Excimer Laser Assisted Non-Occlusive Anastomosis (ELANA) techniqueTM facilitates the construction of an end-to-side anastomosis without temporary occlusion of the recipient artery using a platinum ring and a laser. However, the technical challenge of intracranial micro-sutures remained. Experiments using less sutures eventually resulted in a sutureless ELANA (SELANA) anastomosis. After in vitro and in vivo experiments, the SELANA slide was considered feasible for intracranial use although some concrete improvements, like the inclusion of a clip at the back of the device, were needed. Therefore, the development of an ideal sutureless anastomosis is still ongoing. This process is an evolution rather than a revolution. PMID- 21623325 TI - Neuroprotection in acute ischemic stroke. AB - Acute ischemic stroke, one of the most prominent causes of death and disability in the modern world has attracted an extensive amount of research on its pathophysiology and treatment. There has been significant progress with successful reperfusion treatments associated with improved clinical outcomes, but neuroprotective strategies have failed to show clinical benefit despite promising benchtop results. Unfortunately reperfusion strategies can be employed in a small number of patients who present in the appropriate time window. Neuroprotective strategies may aid in prolonging time windows, thereby potentially increasing the number of eligible patients to benefit from reperfusion treatments. Additional strategies can be used to potentially protect the brain after reperfusion thereby preventing edema and hemorrhage. Previous clinical studies have failed to show benefit likely due to poor patient selection, altering time windows that had shown benefit in bench models and failure to link treatments with reperfusion. PMID- 21623326 TI - Microsurgical technique for previously coiled aneurysms. AB - Since the introduction of Guglielmi detachable coils to treat intracranial aneurysms in 1991, the number of patients undergoing endovascular coiling has continuously risen as well as the number of those residual and recurrent previously coiled aneurysms that necessitate a microsurgical occlusion. Between July 1995 and August 2009 we retrospectively analyzed 81 patients with 82 previously coiled aneurysms treated microsurgically at two Finnish Neurosurgical University Hospitals, Helsinki and Kuopio. Fifty-eight aneurysms (71%) were located at anterior circulation and 24 (29%) at posterior circulation. Fifteen patients were operated on within the first month (early surgery) after coiling, whereas 66 were treated later (late surgery). Complete or partial removal of coils during surgery may facilitate clipping, but is significantly (P<0.001) more difficult to accomplish in late surgery. Removal of coils may also increase the chance for poor outcome. Chance of poor outcome increased also with intraoperative aneurysm rupture, size of the aneurysm and posterior circulation location. Good clinical outcome, three months after surgery, was achieved in 71 patients (88%); four patients were severely disabled, and six patients died (three of them due to poor clinical condition). Complete microsurgical occlusion of the residual previously coiled aneurysm is a high-risk procedure in large and giant aneurysms, and these patients should be referred to a dedicated neurovascular center to minimize surgical complications. Bypass procedures may be the best option for demanding growing lesions, especially those in posterior circulation. PMID- 21623327 TI - Intra-arterial thrombolysis and thrombectomy for acute ischemic stroke: technique and results. AB - Endovascular intraarterial (IA) strategies have emerged as important treatment options for patients with acute ischemic stroke who are ineligible for intravenous (IV) tissue plasminogen activator (tPA) or in whom such therapy has failed. The goal of this article is to provide a comprehensive review of percutaneous IA endovascular techniques aimed at revascularization in the setting of acute ischemic stroke from IA thrombolysis, mechanical thrombectomy, and primary intracranial stenting to retrievable-stent technology. For each modality, we focus on the existing clinical data, including our institutional experience and techniques. PMID- 21623328 TI - Diagnosis of ventriculoperitoneal shunt infection using [F-18]-FDG PET: a case report. AB - Infection of cerebrospinal fluid (CSF) shunts is a common occurrence and can often be difficult to diagnose using standard analysis of shunt fluid. This article presents the first case report on the diagnosis of a CSF shunt infection on FDG PET scan. A 26-year-old female underwent ventriculoperitoneal shunt placement after developing a pseudomeningocele subsequent to a suboccipital craniectomy for Chiari malformation. Two months later, the patient presented with abdominal pain and non-specific symptoms and was found to have a perisplenic abscess for which she was adequately treated. Failure of her symptoms to solve and an initial negative shunt CSF analysis prompted the search for other sources of infection. An FDG PET scan performed a week later found evidence of increase tracer uptake around the distal tip of the catheter and a repeat shunt CSF analysis showed evidence of CSF infection. FDG PET may be useful in diagnosing shunt related infections in case of high clinical suspicion when standard diagnostic modalities fail to diagnose hardware infection. PMID- 21623329 TI - Robotic-assisted laparoendoscopic single-site surgery (R-LESS) in urology: an evidence-based analysis. AB - The objective of this manuscript is to provide an evidence-based analysis of the current status and future perspectives of robotic laparoendoscopic single-site surgery (R-LESS). A PubMed search has been performed for all relevant urological literature regarding natural orifice transluminal endoscopic surgery (NOTES) and laparoendoscopic single-site surgery (LESS). All clinical and investigative reports for robotic LESS and NOTES procedures in the urological literature have been considered. A significant number of clinical urological procedures have been successfully completed utilizing R-LESS procedures. The available experience is limited to referral centers, where the case volume is sufficient to help overcome the challenges and learning curve of LESS surgery. The robotic interface remains the best fit for LESS procedures but its mode of use continues to evolve in attempts to improve surgical technique. We stand today at the dawn of R-LESS surgery, but this approach may well become the standard of care in the near future. Further technological development is needed to allow widespread adoption of the technique. PMID- 21623330 TI - Laparoendoscopic single site (LESS) radical prostatectomy: a review of the initial experience. AB - Surgical treatment for prostate cancer has changed dramatically in recent years due to the incorporation of minimally invasive techniques in the surgical armamentarium. Open surgical approaches to the prostate have largely given way to laparoscopic and robotic techniques. In order to further reduce incisional morbidity and improve cosmesis, there has been a recent interest in laparoendoscopic single site (LESS) approaches to the prostate. Despite a rising interest, there is little available data on these procedures. We performed a systematic review of the literature using MEDLINE, OVID, and Web of Science to identify all publications including LESS radical prostatectomy to date. Manual bibliographic review of cross-referenced items was also performed. We attempt to identify and summarize existing data on these procedures both with and without robotic assistance. Additionally, we review the emerging devices, instruments, cameras, and ports that have made these procedures possible. Next, we offer insight into how this rapidly moving field may transition in the future. Finally, we provide our commentary on this surgical approach, its impact on urology, and how it may help us evolve in the future. PMID- 21623331 TI - Updates on therapeutic targets and agents in castration-resistant prostate cancer. AB - Prostate cancer (PCa) is the most commonly diagnosed noncutaneous cancer in men accounting for 28% of all newly diagnosed cancer cases and it is the second to third most common cause of cancer death in the Western world. Nearly all patients with metastatic disease will eventually experience disease progression despite castration as the median duration of response is between 18-24 months. Hence, development of castration-resistant prostate cancer (CRPC) is only a matter of time in these patients. CRPC is defined by disease progression despite androgen deprivation therapy. CRPC presents a spectrum of disease ranging from rising PSA levels to metastases and significant debilitation from cancer symptoms. Prognosis is associated with several factors, including performance status, presence of bone pain, extent of disease on bone scan, and serum levels of alkaline phosphatase. Based on our enhanced understanding of tumor biology, including the role of tumor, host, and hormonal signaling, there has been rational development of new therapies for CRPC. Over the last decade, several clinical trials have been launched to study novel agents targeting different mechanisms of PCa progression, and have culminated success of new agents for CRPC (docetaxel, cabazitaxel, sipuleucel-T, denosumab, and abiraterone acetate) and several more molecules are on the horizon. The purpose of this review is to discuss the new therapeutic targets in CRPC focusing on new promising agents. PMID- 21623332 TI - Active surveillance for favorable risk prostate cancer: rationale, results, and vis a vis focal therapy role. AB - Active surveillance for Active surveillance (AS), since first being described in 2002 is now an accepted treatment strategy for men with low risk CaP where previously they faced radical whole gland treatment (surgery, external-beam radiation or brachytherapy). AS has built upon the experience of watchful waiting in men believed to not require radical treatment given their age or co morbidities that were both felt to compete with the risk of death from their CaP. AS and radical treatments both have merits and disadvantages. AS has minimal morbidity but the inherent risk of progression associated with expectant management. Radical therapies have an impact on erectile function and continence but provide definitive treatment. Between AS and radical treatment lies focal therapy. Although appearing safe, focal therapy has been limited to small cohorts with short follow-up and cannot be recommended outside study protocols. Most men and their physicians with favorable risk CaP choose between AS and radical therapy. In this review we will focus upon the rationale, patient selection, method of follow-up, triggers for intervention, and the published experience with men undergoing AS with low-risk CaP. We propose a complementary role for surveillance and focal therapy. PMID- 21623333 TI - BK virus associated nephropathy in renal transplantation: where do we stand. AB - BK virus is an increasingly identified complication in renal allograft recipients. During the last decade, the use of potent immunosuppressive medications has led to reemergence of this virus. Despite the paucity of randomized trials, we have come a long way in the knowledge of BK virus associated nephropathy. This review highlights the epidemiological, pathogenic, pathological, and clinical aspects of BK virus. It summarizes advances made in prophylaxis and treatment strategies to curtail this virus in an era of modern immunosuppression. The old word of wisdom- prevention is better than cure- might be relevant in context of BK virus prophylaxis with flouroquinolones in years to follow. PMID- 21623334 TI - Laparoendoscopic single-site and transvaginal nephrectomy. AB - Significant energy has been applied to development of minimally-invasive techniques in surgery to decrease morbidity, improve cosmesis, and hasten postoperative convalescence without compromising surgical outcomes. The evolution of laparoscopy has revolutionized simple, radical and donor nephrectomy in appropriately selected patients, exchanging large flank and abdominal incisions for small "keyhole" incisions. Surgeons continue to pursue innovative approaches to minimize the morbidity of procedures. Natural orifice translumenal endoscopic surgery (NOTES) eliminates entirely the need for abdominal incisions, while laparoendoscopic single-site (LESS) surgery permits placement of multiple instruments through a single incision. Together, they represent two novel approaches developed within the last decade that have been successfully applied to nephrectomy in both the laboratory and clinical settings. Here, the transvaginal and LESS approaches to nephrectomy are reviewed. PMID- 21623335 TI - The use of extra-corporeal shockwave lithotripsy for obstructing ureteric stones. AB - Extracorporeal shock wave lithotripsy (SWL) is a non-invasive treatment for urinary tract stones. This review presents the role of SWL for treating specifically ureteric calculi. The impact of hydronephrosis on SWL success, the use of SWL as emergency treatment (eSWL) for acute ureteric colic, the influence of ureteric stent placement on SWL outcome and the use of medical expulsive therapy to augment SWL success are discussed. PMID- 21623336 TI - The concomitant cryosurgical Cox-Maze procedure using Argon based cryoprobes: 12 month results. AB - AIM: The cut and sew Cox-Maze III procedure has proven to be extremely effective in curing atrial fibrillation. Due to the relative complexity various procedures were developed to apply ablative lesions to treat atrial fibrillation using different energy sources. In this report we present data related to our experience with Argon based cryoablation system in patients having first time concomitant CryoCox-Maze III procedure and other cardiac surgical procedures. METHODS: This is a prospective study where all patients undergoing the Cox maze procedure are entered into our unique maze registry and are followed at 3, 6, 12, 18, 24 months and yearly thereafter. Health related quality of life (SF-12) and atrial fibrillation frequency and severity of symptoms were obtained preoperatively and at follow up. Rhythm was verified by EKG and 24 hour holter. The Heart Rhythm Society definition of failure (any monitored incident of an atrial arrhythmia >30 seconds) was used to compute the rate of return to sinus rhythm. The ablative technique employed was argon based crytothermia using only 1 to 2 atriotomies. RESULTS: The total number of patients operated by multiple surgeons was 124 with 17% through a right minithoracotomy. The operative mortality (<2%) and perioperative stroke rate (<1%) were very low. At 12 months 87% of the patients were in sinus rhythm and off class I and III antriarrhythmic drugs. There was a clear difference in success rate depending on surgeon's total experience. Quality of life and severity of symptoms were improved significantly. CONCLUSION: The one year results of the CryoCox-Maze III procedure when performed concomitantly with another cardiac surgical procedure demonstrate reasonable safety and efficacy. However, operator experience may be related to better outcome. The ablation of atrial fibrillation may be associated with improved quality of life and symptoms relief. PMID- 21623337 TI - Effectiveness of NIRS to sample the frontal brain cortex in all cardiac surgery patients. PMID- 21623338 TI - Renal support. AB - Renal support, intended as a refined and context-sensitive form of severe acute kidney injury management, might be achieved by administering renal replacement therapy with the correct timing and indication, correct prescription and, also, by the expertise and capacity of clinicians to tailor different RRTs to different patients. Furthermore, technical evolution and extended indications for extracorporeal treatments, currently allow the support of multiple organs, other than the isolated kidney failure. Unfortunately, current literature in the field of optimal management of severe acute kidney injury is controversial and lacks a standard of care. This review aims to describe the recent clinical, scientific and technical evolution of renal replacement therapy and the potential suggestive concept of multiple organ support therapy. PMID- 21623340 TI - Ultrasound-guided posterior approach to brachial plexus for the treatment of upper phantom limb syndrome. AB - The purpose of the case is to report the clinical value of the ultrasound-guided posterior approach to the brachial plexus in the treatment of phantom limb syndrome after an upper extremity amputation. The author experienced ultrasound guidance as sole technique to localize the brachial plexus for the purpose of placing a catheter for continuous infusion of a local anesthetic in a patient where standard landmark-based nerve stimulation for placement of a continuous perineural block was not possible. PMID- 21623341 TI - Drowning. AB - Approximately 500,000 deaths due to drowning are reported annually, 30,000 of which are reported in Europe. Because of the relatively low incidence of drowning victims at emergency departments, most emergency physicians do not routinely handle drowning victims. Although confusion regarding the classification and pathophysiology of drowning could be reduced by following the Utstein style consensus, the application of therapeutic modalities and, most important, the estimation of probable prognostic outcomes remain difficult for emergency physicians. This article presents an overview of the classification, pathophysiology, emergency-department treatment and prognostic outcomes of drowning accidents. PMID- 21623342 TI - A randomized control trial of patient-controlled epidural analgesia (PCEA) with and without a background infusion using levobupivacaine and fentanyl. AB - BACKGROUND: Continuous infusion associated with patient-controlled epidural analgesia (PCEA) is used in many maternal units. This randomized controlled study evaluated the effect of a 10 mL/h background infusion associated with a 10 mL-20 minutes lockout time demand-only PCEA protocol using L-bupivacaine plus fentanyl in terms of local anaesthetic consumption, pain management and maternal satisfaction. METHODS: Forty consenting parturients were randomly assigned to receive a 0.125% levobupivacaine plus 1.5 mcg/mL fentanyl PCEA (10 mL bolus with a 20 min lock time) with or without a 10 mL/h background infusion. The total volume of local anesthetic, the number of PCEA demand boluses, pain levels, delivery outcome and maternal satisfaction were evaluated. RESULTS: The total volume of local anaesthetic was 35[20-120] mL in demand-only PCEA group versus 63.8[22.5-123] mL in PCEA plus background infusion group (P<0.001). This decrease in total volume was associated with an increase of self-administrated boluses in demand-only group (3.5[2-12] boluses, versus 1[0-3] bolus in PCEA plus background infusion group) (P<0.001). Pain scores were comparable between groups at any time of the study (P>0.05). Maternal satisfaction did not differ between groups (10[8 10] vs. 10[7-10]; P=0.11). CONCLUSION: When a levobupivacaine plus fentanyl PCEA protocol with high volume boluses and long lockout interval is used for labour analgesia, the background infusion increased the total local anesthetic dose with no change in pain management and maternal satisfaction. PMID- 21623343 TI - Acute physiologic effects of a stepwise recruitment maneuver in acute respiratory distress syndrome. AB - BACKGROUND: Stepwise recruitment maneuvers (RM) applied with high airway pressures may optimize lung recruitment, but this kind of intervention may lead to widely heterogeneous responses with possible side effects. To assess the clinical impact of these maneuvers, we performed a stepwise maximal-recruitment strategy superimposed on routine mechanical ventilation. METHODS: We studied 13 adults with early-phase acute lung injury/acute respiratory distress syndrome (ALI/ARDS). We used pressure-control ventilation at an FiO(2) of 1. Starting from a Pplat/positive end-expiratory pressure (PEEP) of 40/25, we sequentially increased airway pressure in 5 cmH(2)O steps until a PaO(2)/FiO(2) of 350 mmHg or a Pplat/PEEP of 60/40 cmH(2)O was reached. The PEEP was then progressively reduced until the PaO(2) decreased by more than 10% of the maximum PaO(2); the PEEP was subsequently set to 2 cmH(2)O above this level. An intra-arterial catheter continuously displayed blood gas measures. The respiratory mechanics and hemodynamics were monitored at each phase and during the two-hour follow-up. RESULTS: Two hours after the RM, the PaO(2)/FiO(2) was higher than at baseline (187+/-102 versus 339+/-136 mmHg, P<0.001). In 8 patients, the PEEP increased from 12+/-3 cmH(2)O to 15+/-4 cmH(2)O after the RM (P<0.001). In the other five, it closely mirrored the basal PEEP. Seven patients did not reach the 350-mmHg PaO(2) target. The respiratory system compliance decreased in seven patients. The RM was discontinued due to severe complications in four patients. CONCLUSION: Although stepwise-RM improves oxygenation, it has a heterogeneous impact on respiratory mechanics and may cause adverse hemodynamic effects and transient hypoxemia. If the use of this kind of RM is considered, it should be adapted to individual patient needs, applied carefully and closely monitored. PMID- 21623344 TI - An RNA-protein complex links enhanced nuclear 3' processing with cytoplasmic mRNA stabilization. AB - Post-transcriptional controls are critical to gene regulation. These controls are frequently based on sequence-specific binding of trans-acting proteins to cis acting motifs on target RNAs. Prior studies have revealed that the KH-domain protein, alphaCP, binds to a 3' UTR C-rich motif of halpha-globin mRNA and contributes to its cytoplasmic stability. Here, we report that this 3' UTR alphaCP complex regulates the production of mature alpha-globin mRNA by enhancing 3' processing of the halpha-globin transcript. We go on to demonstrate that this nuclear activity reflects enhancement of both the cleavage and the polyadenylation reactions and that alphaCP interacts in vivo with core components of the 3' processing complex. Consistent with its nuclear processing activity, our studies reveal that alphaCP assembles co-transcriptionally at the halpha globin chromatin locus and that this loading is selectively enriched at the 3' terminus of the gene. The demonstrated linkage of nuclear processing with cytoplasmic stabilization via a common RNA-protein complex establishes a basis for integration of sequential controls critical to robust and sustained expression of a target mRNA. PMID- 21623345 TI - The DNA-binding domain of the Chd1 chromatin-remodelling enzyme contains SANT and SLIDE domains. AB - The ATP-dependent chromatin-remodelling enzyme Chd1 is a 168-kDa protein consisting of a double chromodomain, Snf2-related ATPase domain, and a C-terminal DNA-binding domain. Here, we show the DNA-binding domain is required for Saccharomyces cerevisiae Chd1 to bind and remodel nucleosomes. The crystal structure of this domain reveals the presence of structural homology to SANT and SLIDE domains previously identified in ISWI remodelling enzymes. The presence of these domains in ISWI and Chd1 chromatin-remodelling enzymes may provide a means of efficiently harnessing the action of the Snf2-related ATPase domain for the purpose of nucleosome spacing and provide an explanation for partial redundancy between these proteins. Site directed mutagenesis was used to identify residues important for DNA binding and generate a model describing the interaction of this domain with DNA. Through inclusion of Chd1 sequences in homology searches SLIDE domains were identified in CHD6-9 proteins. Point mutations to conserved amino acids within the human CHD7 SLIDE domain have been identified in patients with CHARGE syndrome. PMID- 21623346 TI - A hypomorphic IgH-chain allele affects development of B-cell subsets and favours receptor editing. AB - The quality and quantity of BCR signals impact on cell fate decisions of B lymphocytes. Here, we describe novel gene-targeted mice, which in the context of normal VDJ recombination show hypomorphic expression of immunoglobulin MU heavy chain (MUHC) mRNA levels and hence lower pre-BCR and BCR levels. Hypomorphic expression of MUHC leads to augmented selection processes at all stages of B-cell development, noticeably at the expansion of pre-B cells, the positive selection of immature B lymphocytes in the bone marrow and the selection of the follicular (FO), marginal zone (MZ) and B1 B-lymphocyte compartment in peripheral lymphoid organs. Immature as well as mature FO and MZ B lymphocytes in the peripheral lymphoid organs express lower levels of the receptor for B-cell activating factor (BAFF). In addition, hypomorphic expression of the BCR favours receptor editing. Together, our results highlight the critical importance of pre-BCR and BCR receptor levels for the normal development of B-lymphocyte subpopulations in the context of intact VDJ recombination and a diverse antibody repertoire. PMID- 21623347 TI - Database of NIH grants using machine-learned categories and graphical clustering. PMID- 21623348 TI - Predicting protein associations with long noncoding RNAs. PMID- 21623350 TI - Bringing fly brains in line. PMID- 21623351 TI - Channeling the data deluge. PMID- 21623352 TI - All-in-one live: genes trapped, tagged and conditionally broken. PMID- 21623353 TI - Computational methods for transcriptome annotation and quantification using RNA seq. AB - High-throughput RNA sequencing (RNA-seq) promises a comprehensive picture of the transcriptome, allowing for the complete annotation and quantification of all genes and their isoforms across samples. Realizing this promise requires increasingly complex computational methods. These computational challenges fall into three main categories: (i) read mapping, (ii) transcriptome reconstruction and (iii) expression quantification. Here we explain the major conceptual and practical challenges, and the general classes of solutions for each category. Finally, we highlight the interdependence between these categories and discuss the benefits for different biological applications. PMID- 21623354 TI - De novo genome sequencing and comparative genomics of date palm (Phoenix dactylifera). AB - Date palm is one of the most economically important woody crops cultivated in the Middle East and North Africa and is a good candidate for improving agricultural yields in arid environments. Nonetheless, long generation times (5-8 years) and dioecy (separate male and female trees) have complicated its cultivation and genetic analysis. To address these issues, we assembled a draft genome for a Khalas variety female date palm, the first publicly available resource of its type for a member of the order Arecales. The ~380 Mb sequence, spanning mainly gene-rich regions, includes >25,000 gene models and is predicted to cover ~90% of genes and ~60% of the genome. Sequencing of eight other cultivars, including females of the Deglet Noor and Medjool varieties and their backcrossed males, identified >3.5 million polymorphic sites, including >10,000 genic copy number variations. A small subset of these polymorphisms can distinguish multiple varieties. We identified a region of the genome linked to gender and found evidence that date palm employs an XY system of gender inheritance. PMID- 21623355 TI - Global gene disruption in human cells to assign genes to phenotypes by deep sequencing. AB - Insertional mutagenesis in a haploid background can disrupt gene function. We extend our earlier work by using a retroviral gene-trap vector to generate insertions in >98% of the genes expressed in a human cancer cell line that is haploid for all but one of its chromosomes. We apply phenotypic interrogation via tag sequencing (PhITSeq) to examine millions of mutant alleles through selection and parallel sequencing. Analysis of pools of cells, rather than individual clones enables rapid assessment of the spectrum of genes involved in the phenotypes under study. This facilitates comparative screens as illustrated here for the family of cytolethal distending toxins (CDTs). CDTs are virulence factors secreted by a variety of pathogenic Gram-negative bacteria responsible for tissue damage at distinct anatomical sites. We identify 743 mutations distributed over 12 human genes important for intoxication by four different CDTs. Although related CDTs may share host factors, they also exploit unique host factors to yield a profile characteristic for each CDT. PMID- 21623356 TI - Kinase inhibitors modulate huntingtin cell localization and toxicity. AB - Two serine residues within the first 17 amino acid residues of huntingtin (N17) are crucial for modulation of mutant huntingtin toxicity in cell and mouse genetic models of Huntington's disease. Here we show that the stress-dependent phosphorylation of huntingtin at Ser13 and Ser16 affects N17 conformation and targets full-length huntingtin to chromatin-dependent subregions of the nucleus, the mitotic spindle and cleavage furrow during cell division. Polyglutamine expanded mutant huntingtin is hypophosphorylated in N17 in both homozygous and heterozygous cell contexts. By high-content screening in live cells, we identified kinase inhibitors that modulated N17 phosphorylation and hence huntingtin subcellular localization. N17 phosphorylation was reduced by casein kinase-2 inhibitors. Paradoxically, IKKbeta kinase inhibition increased N17 phosphorylation, affecting huntingtin nuclear and subnuclear localization. These data indicate that huntingtin phosphorylation at Ser13 and Ser16 can be modulated by small-molecule drugs, which may have therapeutic potential in Huntington's disease. PMID- 21623357 TI - FrsA functions as a cofactor-independent decarboxylase to control metabolic flux. AB - The interaction between fermentation-respiration switch (FrsA) protein and glucose-specific enzyme IIA(Glc) increases glucose fermentation under oxygen limited conditions. We show that FrsA converts pyruvate to acetaldehyde and carbon dioxide in a cofactor-independent manner and that its pyruvate decarboxylation activity is enhanced by the dephosphorylated form of IIA(Glc) (d IIA(Glc)). Crystal structures of FrsA and its complex with d-IIA(Glc) revealed residues required for catalysis as well as the structural basis for the activation by d-IIA(Glc). PMID- 21623358 TI - Origin and role of distal visceral endoderm, a group of cells that determines anterior-posterior polarity of the mouse embryo. AB - Anterior-posterior polarity of the mouse embryo has been thought to be established when distal visceral endoderm (DVE) at embryonic day (E) 5.5 migrates toward the future anterior side to form anterior visceral endoderm (AVE). Lefty1, a marker of DVE and AVE, is asymmetrically expressed in implanting mouse embryos. We now show that Lefty1 is expressed first in a subset of epiblast progenitor cells and then in a subset of primitive endoderm progenitors. Genetic fate mapping indicated that the latter cells are destined to become DVE. In contrast to the accepted notion, however, AVE is not derived from DVE but is newly formed after E5.5 from Lefty1(-) visceral endoderm cells that move to the distal tip. Concomitant with DVE migration, all visceral endoderm cells in the embryonic region undergo global movement. In embryos subjected to genetic ablation of Lefty1-expressing DVE cells, AVE was formed de novo but the visceral endoderm including the newly formed AVE failed to migrate, indicating that DVE guides the migration of AVE by initiating the global movement of visceral endoderm cells. Future anterior-posterior polarity is thus already determined by Lefty1(+) blastomeres in the implanting blastocyst. PMID- 21623359 TI - Cold-atom scanning probe microscopy. AB - Scanning probe microscopes are widely used to study surfaces with atomic resolution in many areas of nanoscience. Ultracold atomic gases trapped in electromagnetic potentials can be used to study electromagnetic interactions between the atoms and nearby surfaces in chip-based systems. Here we demonstrate a new type of scanning probe microscope that combines these two areas of research by using an ultracold gas as the tip in a scanning probe microscope. This cold atom scanning probe microscope offers a large scanning volume, an ultrasoft tip of well-defined shape and high purity, and sensitivity to electromagnetic forces (including dispersion forces near nanostructured surfaces). We use the cold-atom scanning probe microscope to non-destructively measure the position and height of carbon nanotube structures and individual free-standing nanotubes. Cooling the atoms in the gas to form a Bose-Einstein condensate increases the resolution of the device. PMID- 21623360 TI - Highly uniform and reproducible surface-enhanced Raman scattering from DNA tailorable nanoparticles with 1-nm interior gap. AB - An ideal surface-enhanced Raman scattering (SERS) nanostructure for sensing and imaging applications should induce a high signal enhancement, generate a reproducible and uniform response, and should be easy to synthesize. Many SERS active nanostructures have been investigated, but they suffer from poor reproducibility of the SERS-active sites, and the wide distribution of their enhancement factor values results in an unquantifiable SERS signal. Here, we show that DNA on gold nanoparticles facilitates the formation of well-defined gold nanobridged nanogap particles (Au-NNP) that generate a highly stable and reproducible SERS signal. The uniform and hollow gap (~1 nm) between the gold core and gold shell can be precisely loaded with a quantifiable amount of Raman dyes. SERS signals generated by Au-NNPs showed a linear dependence on probe concentration (R(2) > 0.98) and were sensitive down to 10 fM concentrations. Single-particle nano-Raman mapping analysis revealed that >90% of Au-NNPs had enhancement factors greater than 1.0 * 10(8), which is sufficient for single molecule detection, and the values were narrowly distributed between 1.0 * 10(8) and 5.0 * 10(9). PMID- 21623361 TI - Synaptic vesicle retrieval time is a cell-wide rather than individual-synapse property. AB - Although individual nerve terminals from the same neuron often differ in neurotransmitter release characteristics, the extent to which endocytic retrieval of synaptic vesicle components differs is unknown. We used high-fidelity optical recordings to undertake a large-scale analysis of endocytosis kinetics of individual boutons in hippocampal rat neurons. Our data indicate that endocytosis kinetics do not differ substantially across boutons from the same cell but instead appear to be controlled at a cell-wide level. PMID- 21623362 TI - Differential coupling of visual cortex with default or frontal-parietal network based on goals. AB - The relationship between top-down enhancement and suppression of sensory cortical activity and large-scale neural networks remains unclear. Functional connectivity analysis of human functional magnetic resonance imaging data revealed that visual cortical areas that selectively process relevant information are functionally connected with the frontal-parietal network, whereas those that process irrelevant information are simultaneously coupled with the default network. This indicates that sensory cortical regions are differentially and dynamically coupled with distinct networks on the basis of task goals. PMID- 21623363 TI - Distinct functions of kainate receptors in the brain are determined by the auxiliary subunit Neto1. AB - Ionotropic glutamate receptors principally mediate fast excitatory transmission in the brain. Among the three classes of ionotropic glutamate receptors, kainate receptors (KARs) have a unique brain distribution, which has been historically defined by (3)H-radiolabeled kainate binding. Compared with recombinant KARs expressed in heterologous cells, synaptic KARs exhibit characteristically slow rise-time and decay kinetics. However, the mechanisms responsible for these distinct KAR properties remain unclear. We found that both the high-affinity binding pattern in the mouse brain and the channel properties of native KARs are determined by the KAR auxiliary subunit Neto1. Through modulation of agonist binding affinity and off-kinetics of KARs, but not trafficking of KARs, Neto1 determined both the KAR high-affinity binding pattern and the distinctively slow kinetics of postsynaptic KARs. By regulating KAR excitatory postsynaptic current kinetics, Neto1 can control synaptic temporal summation, spike generation and fidelity. PMID- 21623365 TI - Retinal origin of orientation maps in visual cortex. AB - The orientation map is a hallmark of primary visual cortex in higher mammals. It is not yet known how orientation maps develop, what function they have in visual processing and why some species lack them. Here we advance the notion that quasi periodic orientation maps are established by moire interference of regularly spaced ON- and OFF-center retinal ganglion cell mosaics. A key prediction of the theory is that the centers of iso-orientation domains must be arranged in a hexagonal lattice on the cortical surface. Here we show that such a pattern is observed in individuals of four different species: monkeys, cats, tree shrews and ferrets. The proposed mechanism explains how orientation maps can develop without requiring precise patterns of spontaneous activity or molecular guidance. Further, it offers a possible account for the emergence of orientation tuning in single neurons despite the absence of orderly orientation maps in rodents species. PMID- 21623364 TI - Rapid activity-induced transcription of Arc and other IEGs relies on poised RNA polymerase II. AB - Transcription of immediate early genes (IEGs) in neurons is highly sensitive to neuronal activity, but the mechanism underlying these early transcription events is largely unknown. We found that several IEGs, such as Arc (also known as Arg3.1), are poised for near-instantaneous transcription by the stalling of RNA polymerase II (Pol II) just downstream of the transcription start site in rat neurons. Depletion through RNA interference of negative elongation factor, a mediator of Pol II stalling, reduced the Pol II occupancy of the Arc promoter and compromised the rapid induction of Arc and other IEGs. In contrast, reduction of Pol II stalling did not prevent transcription of IEGs that were expressed later and largely lacked promoter-proximal Pol II stalling. Together, our data strongly indicate that the rapid induction of neuronal IEGs requires poised Pol II and suggest a role for this mechanism in a wide variety of transcription-dependent processes, including learning and memory. PMID- 21623366 TI - The nucleosome map of the mammalian liver. AB - Binding to nucleosomal DNA is critical for 'pioneer' transcription factors such as the winged-helix transcription factors Foxa1 and Foxa2 to regulate chromatin structure and gene activation. Here we report the genome-wide map of nucleosome positions in the mouse liver, with emphasis on transcriptional start sites, CpG islands, Foxa2 binding sites and their correlation with gene expression. Despite the heterogeneity of liver tissue, we could clearly discern the nucleosome pattern of the predominant liver cell, the hepatocyte. By analyzing nucleosome occupancy and the distributions of heterochromatin protein 1 (Hp1), CBP (also known as Crebbp) and p300 (Ep300) in Foxa1- and Foxa2-deficient livers, we find that the maintenance of nucleosome position and chromatin structure surrounding Foxa2 binding sites is independent of Foxa1 and Foxa2. PMID- 21623367 TI - Structure of the no-go mRNA decay complex Dom34-Hbs1 bound to a stalled 80S ribosome. AB - No-go decay (NGD) is a mRNA quality-control mechanism in eukaryotic cells that leads to degradation of mRNAs stalled during translational elongation. The key factors triggering NGD are Dom34 and Hbs1. We used cryo-EM to visualize NGD intermediates resulting from binding of the Dom34-Hbs1 complex to stalled ribosomes. At subnanometer resolution, all domains of Dom34 and Hbs1 were identified, allowing the docking of crystal structures and homology models. Moreover, the close structural similarity of Dom34 and Hbs1 to eukaryotic release factors (eRFs) enabled us to propose a model for the ribosome-bound eRF1-eRF3 complex. Collectively, our data provide structural insights into how stalled mRNA is recognized on the ribosome and how the eRF complex can simultaneously recognize stop codons and catalyze peptide release. PMID- 21623368 TI - Single-molecule fluorescence reveals sequence-specific misfolding in multidomain proteins. AB - A large range of debilitating medical conditions is linked to protein misfolding, which may compete with productive folding particularly in proteins containing multiple domains. Seventy-five per cent of the eukaryotic proteome consists of multidomain proteins, yet it is not understood how interdomain misfolding is avoided. It has been proposed that maintaining low sequence identity between covalently linked domains is a mechanism to avoid misfolding. Here we use single molecule Forster resonance energy transfer to detect and quantify rare misfolding events in tandem immunoglobulin domains from the I band of titin under native conditions. About 5.5 per cent of molecules with identical domains misfold during refolding in vitro and form an unexpectedly stable state with an unfolding half time of several days. Tandem arrays of immunoglobulin-like domains in humans show significantly lower sequence identity between neighbouring domains than between non-adjacent domains. In particular, the sequence identity of neighbouring domains has been found to be preferentially below 40 per cent. We observe no misfolding for a tandem of naturally neighbouring domains with low sequence identity (24 per cent), whereas misfolding occurs between domains that are 42 per cent identical. Coarse-grained molecular simulations predict the formation of domain-swapped structures that are in excellent agreement with the observed transfer efficiency of the misfolded species. We infer that the interactions underlying misfolding are very specific and result in a sequence-specific domain swapping mechanism. Diversifying the sequence between neighbouring domains seems to be a successful evolutionary strategy to avoid misfolding in multidomain proteins. PMID- 21623369 TI - Regulation of angiogenesis by a non-canonical Wnt-Flt1 pathway in myeloid cells. AB - Myeloid cells are a feature of most tissues. Here we show that during development, retinal myeloid cells (RMCs) produce Wnt ligands to regulate blood vessel branching. In the mouse retina, where angiogenesis occurs postnatally, somatic deletion in RMCs of the Wnt ligand transporter Wntless results in increased angiogenesis in the deeper layers. We also show that mutation of Wnt5a and Wnt11 results in increased angiogenesis and that these ligands elicit RMC responses via a non-canonical Wnt pathway. Using cultured myeloid-like cells and RMC somatic deletion of Flt1, we show that an effector of Wnt-dependent suppression of angiogenesis by RMCs is Flt1, a naturally occurring inhibitor of vascular endothelial growth factor (VEGF). These findings indicate that resident myeloid cells can use a non-canonical, Wnt-Flt1 pathway to suppress angiogenic branching. PMID- 21623370 TI - Latent TGF-beta binding protein 3 identifies a second heart field in zebrafish. AB - The four-chambered mammalian heart develops from two fields of cardiac progenitor cells distinguished by their spatiotemporal patterns of differentiation and contributions to the definitive heart. The first heart field differentiates earlier in lateral plate mesoderm, generates the linear heart tube and ultimately gives rise to the left ventricle. The second heart field (SHF) differentiates later in pharyngeal mesoderm, elongates the heart tube, and gives rise to the outflow tract and much of the right ventricle. Because hearts in lower vertebrates contain a rudimentary outflow tract but not a right ventricle, the existence and function of SHF-like cells in these species has remained a topic of speculation. Here we provide direct evidence from Cre/Lox-mediated lineage tracing and loss-of-function studies in zebrafish, a lower vertebrate with a single ventricle, that latent TGF-beta binding protein 3 (ltbp3) transcripts mark a field of cardiac progenitor cells with defining characteristics of the anterior SHF in mammals. Specifically, ltbp3(+) cells differentiate in pharyngeal mesoderm after formation of the heart tube, elongate the heart tube at the outflow pole, and give rise to three cardiovascular lineages in the outflow tract and myocardium in the distal ventricle. In addition to expressing Ltbp3, a protein that regulates the bioavailability of TGF-beta ligands, zebrafish SHF cells co express nkx2.5, an evolutionarily conserved marker of cardiac progenitor cells in both fields. Embryos devoid of ltbp3 lack the same cardiac structures derived from ltbp3(+) cells due to compromised progenitor proliferation. Furthermore, small-molecule inhibition of TGF-beta signalling phenocopies the ltbp3-morphant phenotype whereas expression of a constitutively active TGF-beta type I receptor rescues it. Taken together, our findings uncover a requirement for ltbp3-TGF-beta signalling during zebrafish SHF development, a process that serves to enlarge the single ventricular chamber in this species. PMID- 21623371 TI - A gene regulatory network controlling the embryonic specification of endoderm. AB - Specification of endoderm is the prerequisite for gut formation in the embryogenesis of bilaterian organisms. Modern lineage labelling studies have shown that in the sea urchin embryo model system, descendants of the veg1 and veg2 cell lineages produce the endoderm, and that the veg2 lineage also gives rise to mesodermal cell types. It is known that Wnt/beta-catenin signalling is required for endoderm specification and Delta/Notch signalling is required for mesoderm specification. Some direct cis-regulatory targets of these signals have been found and various phenomenological patterns of gene expression have been observed in the pre-gastrular endomesoderm. However, no comprehensive, causal explanation of endoderm specification has been conceived for sea urchins, nor for any other deuterostome. Here we propose a model, on the basis of the underlying genomic control system, that provides such an explanation, built at several levels of biological organization. The hardwired core of the control system consists of the cis-regulatory apparatus of endodermal regulatory genes, which determine the relationship between the inputs to which these genes are exposed and their outputs. The architecture of the network circuitry controlling the dynamic process of endoderm specification then explains, at the system level, a sequence of developmental logic operations, which generate the biological process. The control system initiates non-interacting endodermal and mesodermal gene regulatory networks in veg2-derived cells and extinguishes the endodermal gene regulatory network in mesodermal precursors. It also generates a cross regulatory network that specifies future anterior endoderm in veg2 descendants and institutes a distinct network specifying posterior endoderm in veg1-derived cells. The network model provides an explanatory framework that relates endoderm specification to the genomic regulatory code. PMID- 21623372 TI - An integrated approach to characterize genetic interaction networks in yeast metabolism. AB - Although experimental and theoretical efforts have been applied to globally map genetic interactions, we still do not understand how gene-gene interactions arise from the operation of biomolecular networks. To bridge the gap between empirical and computational studies, we i, quantitatively measured genetic interactions between ~185,000 metabolic gene pairs in Saccharomyces cerevisiae, ii, superposed the data on a detailed systems biology model of metabolism and iii, introduced a machine-learning method to reconcile empirical interaction data with model predictions. We systematically investigated the relative impacts of functional modularity and metabolic flux coupling on the distribution of negative and positive genetic interactions. We also provide a mechanistic explanation for the link between the degree of genetic interaction, pleiotropy and gene dispensability. Last, we show the feasibility of automated metabolic model refinement by correcting misannotations in NAD biosynthesis and confirming them by in vivo experiments. PMID- 21623373 TI - The splicing regulator Rbfox1 (A2BP1) controls neuronal excitation in the mammalian brain. AB - The Rbfox family of RNA binding proteins regulates alternative splicing of many important neuronal transcripts, but its role in neuronal physiology is not clear. We show here that central nervous system-specific deletion of the gene encoding Rbfox1 results in heightened susceptibility to spontaneous and kainic acid induced seizures. Electrophysiological recording revealed a corresponding increase in neuronal excitability in the dentate gyrus of the knockout mice. Whole-transcriptome analyses identified multiple splicing changes in the Rbfox1( /-) brain with few changes in overall transcript abundance. These splicing changes alter proteins that mediate synaptic transmission and membrane excitation. Thus, Rbfox1 directs a genetic program required in the prevention of neuronal hyperexcitation and seizures. The Rbfox1 knockout mice provide a new model to study the post-transcriptional regulation of synaptic function. PMID- 21623374 TI - Subspecific origin and haplotype diversity in the laboratory mouse. AB - Here we provide a genome-wide, high-resolution map of the phylogenetic origin of the genome of most extant laboratory mouse inbred strains. Our analysis is based on the genotypes of wild-caught mice from three subspecies of Mus musculus. We show that classical laboratory strains are derived from a few fancy mice with limited haplotype diversity. Their genomes are overwhelmingly Mus musculus domesticus in origin, and the remainder is mostly of Japanese origin. We generated genome-wide haplotype maps based on identity by descent from fancy mice and show that classical inbred strains have limited and non-randomly distributed genetic diversity. In contrast, wild-derived laboratory strains represent a broad sampling of diversity within M. musculus. Intersubspecific introgression is pervasive in these strains, and contamination by laboratory stocks has played a role in this process. The subspecific origin, haplotype diversity and identity by descent maps can be visualized using the Mouse Phylogeny Viewer (see URLs). PMID- 21623376 TI - Direct tomography with chemical-bond contrast. AB - Three-dimensional (3D) X-ray imaging methods have advanced tremendously during recent years. Traditional tomography uses absorption as the contrast mechanism, but for many purposes its sensitivity is limited. The introduction of diffraction, small-angle scattering, refraction, and phase contrasts has increased the sensitivity, especially in materials composed of light elements (for example, carbon and oxygen). X-ray spectroscopy, in principle, offers information on element composition and chemical environment. However, its application in 3D imaging over macroscopic length scales has not been possible for light elements. Here we introduce a new hard-X-ray spectroscopic tomography with a unique sensitivity to light elements. In this method, dark-field section images are obtained directly without any reconstruction algorithms. We apply the method to acquire the 3D structure and map the chemical bonding in selected samples relevant to materials science. The novel aspects make this technique a powerful new imaging tool, with an inherent access to the molecular-level chemical environment. PMID- 21623375 TI - Genome-wide association identifies three new susceptibility loci for Paget's disease of bone. AB - Paget's disease of bone (PDB) is a common disorder characterized by focal abnormalities of bone remodeling. We previously identified variants at the CSF1, OPTN and TNFRSF11A loci as risk factors for PDB by genome-wide association study. Here we extended this study, identified three new loci and confirmed their association with PDB in 2,215 affected individuals (cases) and 4,370 controls from seven independent populations. The new associations were with rs5742915 within PML on 15q24 (odds ratio (OR) = 1.34, P = 1.6 * 10(-14)), rs10498635 within RIN3 on 14q32 (OR = 1.44, P = 2.55 * 10(-11)) and rs4294134 within NUP205 on 7q33 (OR = 1.45, P = 8.45 * 10(-10)). Our data also confirmed the association of TM7SF4 (rs2458413, OR = 1.40, P = 7.38 * 10(-17)) with PDB. These seven loci explained ~13% of the familial risk of PDB. These studies provide new insights into the genetic architecture and pathophysiology of PDB. PMID- 21623377 TI - Dirac cones induced by accidental degeneracy in photonic crystals and zero refractive-index materials. AB - A zero-refractive-index metamaterial is one in which waves do not experience any spatial phase change, and such a peculiar material has many interesting wave manipulating properties. These materials can in principle be realized using man made composites comprising metallic resonators or chiral inclusions, but metallic components have losses that compromise functionality at high frequencies. It would be highly desirable if we could achieve a zero refractive index using dielectrics alone. Here, we show that by employing accidental degeneracy, dielectric photonic crystals can be designed and fabricated that exhibit Dirac cone dispersion at the centre of the Brillouin zone at a finite frequency. In addition to many interesting properties intrinsic to a Dirac cone dispersion, we can use effective medium theory to relate the photonic crystal to a material with effectively zero permittivity and permeability. We then numerically and experimentally demonstrate in the microwave regime that such dielectric photonic crystals with reasonable dielectric constants manipulate waves as if they had near-zero refractive indices at and near the Dirac point frequency. PMID- 21623378 TI - Structural origin of enhanced slow dynamics near a wall in glass-forming systems. AB - Spatial confinement is known to induce a drastic change in the viscosity, relaxation times, and flow profile of liquids near the glass (or jamming) transition point. The essential underlying question is how a wall affects the dynamics of densely packed systems. Here we study this fundamental problem, using experiments on a driven granular hard-sphere liquid and numerical simulations of polydisperse and bidisperse colloidal liquids. The nearly hard-core nature of the particle-wall interaction provides an ideal opportunity to study purely geometrical confinement effects. We reveal that the slower dynamics near a wall is induced by wall-induced enhancement of 'glassy structural order', which is a manifestation of strong interparticle correlations. By generalizing the structure dynamics relation for bulk systems, we find a quantitative relation between the structural relaxation time at a certain distance from a wall and the correlation length of glassy structural order there. Our finding suggests that glassy structural ordering may be the origin of the slow glassy dynamics of a supercooled liquid. PMID- 21623379 TI - Innate lymphoid cells mediate influenza-induced airway hyper-reactivity independently of adaptive immunity. AB - Patients with asthma, a major public health problem, are at high risk for serious disease from influenza virus infection, but the pathogenic mechanisms by which influenza A causes airway disease and asthma are not fully known. We show here in a mouse model that influenza infection acutely induced airway hyper-reactivity (AHR), a cardinal feature of asthma, independently of T helper type 2 (T(H)2) cells and adaptive immunity. Instead, influenza infection induced AHR through a previously unknown pathway that required the interleukin 13 (IL-13)-IL-33 axis and cells of the non-T cell, non-B cell innate lymphoid type called 'natural helper cells'. Infection with influenza A virus, which activates the NLRP3 inflammasome, resulted in much more production of IL-33 by alveolar macrophages, which in turn activated natural helper cells producing substantial IL-13. PMID- 21623380 TI - Transcription factor T-bet represses expression of the inhibitory receptor PD-1 and sustains virus-specific CD8+ T cell responses during chronic infection. AB - T cell exhaustion has a major role in failure to control chronic infection. High expression of inhibitory receptors, including PD-1, and the inability to sustain functional T cell responses contribute to exhaustion. However, the transcriptional control of these processes remains unclear. Here we demonstrate that the transcription factor T-bet regulated the exhaustion of CD8(+) T cells and the expression of inhibitory receptors. T-bet directly repressed transcription of the gene encoding PD-1 and resulted in lower expression of other inhibitory receptors. Although a greater abundance of T-bet promoted terminal differentiation after acute infection, high T-bet expression sustained exhausted CD8(+) T cells and repressed the expression of inhibitory receptors during chronic viral infection. Persistent antigenic stimulation caused downregulation of T-bet, which resulted in more severe exhaustion of CD8(+) T cells. Our observations suggest therapeutic opportunities involving higher T-bet expression during chronic infection. PMID- 21623381 TI - Misregulated alternative splicing of BIN1 is associated with T tubule alterations and muscle weakness in myotonic dystrophy. AB - Myotonic dystrophy is the most common muscular dystrophy in adults and the first recognized example of an RNA-mediated disease. Congenital myotonic dystrophy (CDM1) and myotonic dystrophy of type 1 (DM1) or of type 2 (DM2) are caused by the expression of mutant RNAs containing expanded CUG or CCUG repeats, respectively. These mutant RNAs sequester the splicing regulator Muscleblind-like 1 (MBNL1), resulting in specific misregulation of the alternative splicing of other pre-mRNAs. We found that alternative splicing of the bridging integrator-1 (BIN1) pre-mRNA is altered in skeletal muscle samples of people with CDM1, DM1 and DM2. BIN1 is involved in tubular invaginations of membranes and is required for the biogenesis of muscle T tubules, which are specialized skeletal muscle membrane structures essential for excitation-contraction coupling. Mutations in the BIN1 gene cause centronuclear myopathy, which shares some histopathological features with myotonic dystrophy. We found that MBNL1 binds the BIN1 pre-mRNA and regulates its alternative splicing. BIN1 missplicing results in expression of an inactive form of BIN1 lacking phosphatidylinositol 5-phosphate-binding and membrane-tubulating activities. Consistent with a defect of BIN1, muscle T tubules are altered in people with myotonic dystrophy, and membrane structures are restored upon expression of the normal splicing form of BIN1 in muscle cells of such individuals. Finally, reproducing BIN1 splicing alteration in mice is sufficient to promote T tubule alterations and muscle weakness, a predominant feature of myotonic dystrophy. PMID- 21623382 TI - Defective Wnt-dependent cerebellar midline fusion in a mouse model of Joubert syndrome. AB - The ciliopathy Joubert syndrome is marked by cerebellar vermis hypoplasia, a phenotype for which the pathogenic mechanism is unclear. To investigate Joubert syndrome pathogenesis, we have examined mice with mutated Ahi1, the first identified Joubert syndrome-associated gene. These mice show cerebellar hypoplasia with a vermis-midline fusion defect early in development. This defect is concomitant with expansion of the roof plate and is also evident in a mouse mutant for another Joubert syndrome-associated gene, Cep290. Furthermore, fetal magnetic resonance imaging (MRI) of human subjects with Joubert syndrome reveals a similar midline cleft, suggesting parallel pathogenic mechanisms. Previous evidence has suggested a role for Jouberin (Jbn), the protein encoded by Ahi1, in canonical Wnt signaling. Consistent with this, we found decreased Wnt reporter activity at the site of hemisphere fusion in the developing cerebellum of Ahi1 mutant mice. This decrease was accompanied by reduced proliferation at the site of fusion. Finally, treatment with lithium, a Wnt pathway agonist, partially rescued this phenotype. Our findings implicate a defect in Wnt signaling in the cerebellar midline phenotype seen in Joubert syndrome that can be overcome with Wnt stimulation. PMID- 21623383 TI - Prognostic significance of FOXL2 mutation and mRNA expression in adult and juvenile granulosa cell tumors of the ovary. AB - Recently, mutation of the FOXL2 gene has been consistently identified in adult granulosa cell tumors of the ovary. The purpose of this study is to investigate whether the FOXL2 mutation and mRNA expression have a role in the pathogenesis of juvenile and adult granulosa cell tumors and influence tumor progression. Thirty four adult granulosa cell tumors and 20 juvenile granulosa cell tumors were examined for the presence of the FOXL2 (C402G) mutation. Expression levels were studied by quantitative PCR and immunohistochemistry. We found that FOXL2 (C402G) mutation was present in 19/27 (70%) of the adult type tumors but in none of the juvenile granulosa cell tumors (0/18). No correlation was encountered between the presence of FOXL2 mutation and various clinicopathologic parameters except for the presence of a different sex-cord component, which was more frequently found in the subgroup of wild-type adult granulosa cell tumors than in the mutated tumors. Patients with tumors harboring the FOXL2 (C402G) mutation had a worse disease-free survival than those with the wild-type gene. Expression levels of FOXL2 mRNA had an impact on disease-free survival in both adult and juvenile granulosa cell tumors. We also found that the mutated tumors had a higher immunohistochemical expression of the FOXL2 protein, and there was a linear correlation between mRNA and immunohistochemical FOXL2 expression in both adult and juvenile granulosa cell tumors. Patients with juvenile granulosa cell tumors and higher FOXL2 protein expression had worse overall survival and disease-free survival than those with negative or weakly immunoreactive tumors. Our data suggest that FOXL2 mutation and mRNA expression are of prognostic importance in both adult and juvenile granulosa cell tumors. PMID- 21623384 TI - Immunohistochemical algorithm for differentiation of lung adenocarcinoma and squamous cell carcinoma based on large series of whole-tissue sections with validation in small specimens. AB - Immunohistochemistry is increasingly utilized to differentiate lung adenocarcinoma and squamous cell carcinoma. However, detailed analysis of coexpression profiles of commonly used markers in large series of whole-tissue sections is lacking. Furthermore, the optimal diagnostic algorithm, particularly the minimal-marker combination, is not firmly established. We therefore studied whole-tissue sections of resected adenocarcinoma and squamous cell carcinoma (n=315) with markers commonly used to identify adenocarcinoma (TTF-1) and squamous cell carcinoma (p63, CK5/6, 34betaE12), and prospectively validated the devised algorithm in morphologically unclassifiable small biopsy/cytology specimens (n=38). Analysis of whole-tissue sections showed that squamous cell carcinoma had a highly consistent immunoprofile (TTF-1-negative and p63/CK5/6/34betaE12-diffuse) with only rare variation. In contrast, adenocarcinoma showed significant immunoheterogenetity for all 'squamous markers' (p63 (32%), CK5/6 (18%), 34betaE12 (82%)) and TTF-1 (89%). As a single marker, only diffuse TTF-1 was specific for adenocarcinoma whereas none of the 'squamous markers,' even if diffuse, were entirely specific for squamous cell carcinoma. In contrast, coexpression profiles of TTF-1/p63 had only minimal overlap between adenocarcinoma and squamous cell carcinoma, and there was no overlap if CK5/6 was added as a third marker. An algorithm was devised in which TTF-1/p63 were used as the first-line panel, and CK5/6 was added for rare indeterminate cases. Prospective validation of this algorithm in small specimens showed 100% accuracy of adenocarcinoma vs squamous cell carcinoma prediction as determined by subsequent resection. In conclusion, although reactivity for 'squamous markers' is common in lung adenocarcinoma, a two-marker panel of TTF-1/p63 is sufficient for subtyping of the majority of tumors as adenocarcinomas vs squamous cell carcinoma, and addition of CK5/6 is needed in only a small subset of cases. This simple algorithm achieves excellent accuracy in small specimens while conserving the tissue for potential predictive marker testing, which is now an essential consideration in advanced lung cancer specimens. PMID- 21623385 TI - Cytoplasmic p63 immunohistochemistry is a useful marker for muscle differentiation: an immunohistochemical and immunoelectron microscopic study. AB - TP63, a member of the TP53 gene family, is a nuclear marker of myoepithelial cells. Antibody against p63 is frequently used to aid in the diagnosis of prostate carcinoma, as well as in the identification of myoepithelial cells in other tissues including the breast. p63 is also a marker for squamous cell carcinoma. Recently, it was found that all p53 family members are involved in regulating the process of muscle differentiation through the retinoblastoma (RB) protein. Ablation of these p53 family functions blocks the differentiation program and promotes malignant transformation by enabling cooperating oncogenes to transform myoblasts. We therefore studied p63 expression in a number of neoplasms with myogenic differentiation. Immunohistochemical staining for p63 was performed on paraffin sections from 38 rhabdomyosarcomas, five leiomyomas, five leiomyosarcomas, five rhabdomyomas, five rhabdomyomatous Wilms tumors, three normal cardiac muscles, one medullomyoblastoma, one pleuropulmonary blastoma with rhabdomyomatous differentiation, and one teratoma with prominent rhabdomyoblasts. Each case was also stained with desmin. Unlike the nuclear staining scored in myoepithelial cells, only cytoplasmic staining for p63 was considered positive. Of 38 cases of rhabdomyosarcoma, 36 showed cytoplasmic p63 staining; 24 of these showed highlighting of cross-striations superior to that of desmin. In addition, 5/5 rhabdomyomas, 5/5 rhabdomyomatous Wilms tumors, 1/1 pleuropulmonary blastoma with rhabdomyomatous differentiation, 1/1 teratoma with atypical rhabdoblasts, and 1/1 medullomyoblastoma exhibited cytoplasmic p63 staining. Normal cardiac muscle samples (3/3) also demonstrated positive cytoplasmic staining and distinct cross-striations. Smooth muscle tumors exhibited only very focal and faint cytoplasmic staining in 5/5 leiomyomas and 4/5 leiomyosarcomas. Immunoelectron microscopic study of skeletal muscle showed p63 localization to the Z bands of sarcomeres. We conclude that p63 immunostain is a sensitive marker for skeletal muscle differentiation and highlights the cross-striations of strap cells with exceptional definition. PMID- 21623386 TI - Sperm donation and its application in China: a 7-year multicenter retrospective study. AB - Sperm donation in China is different from that in other countries due to cultural, social and political factors. This research presents the current status of sperm donation in Mainland China and highlights some problems. Between January 2003 and December 2009, 19 471 sperm donors were screened totally and 6467 donors (33.2%) were recruited. The primary reasons for non-recruitment were either inadequate semen parameters (55.0%) or positive results for sexually transmitted diseases (7.9%). There were 327 (1.7%) qualified donors who withdrew from the program because of frustration related to failed semen parameters, participation merely for free medical tests or job transfer. A questionnaire investigating donor intention, as well as other concerns associated with sperm donation, was distributed to 516 potential donors. All potential donors indicated their primary motivation as altruism, while 90.9% mentioned monetary reward as a second motivating factor. Approximately 93.4% of donors expressed some apprehension about the risk of consanguineous mating and the protection of their identity. Over the past 7 years, 488 389 vials of donors' semen have been cryopreserved. In 36 438 artificial insemination with donor sperm (AID) cycles, the clinical pregnancy rate was 23.9% and the live birth rate was 16.6%. In 7148 in vitro fertilization cycles, the clinical pregnancy rate was 45.8% and the live birth rate was 35.2%. Human sperm banks have been strictly monitored to ensure that each sperm donor can only impregnate five women nationwide. There is still a large gap between the supply and demand for sperm donation which may be solved by updated guidelines. PMID- 21623387 TI - TRP channels in prostate cancer: the good, the bad and the ugly? AB - During the last decade, transient receptor potential (TRP) channels emerge as key proteins in central mechanisms of the carcinogenesis such as cell proliferation, apoptosis and migration. Initial studies showed that expression profile of some TRP channels, notably TRP melastatin 8 (TRPM8), TRP vanilloid 6 (TRPV6),TRP canonical (TRPC6) and TRPV2, is changing during the development and the progression of prostate cancer towards the hormone-refractory stages. The link between the change in expression levels and the functional role of these channels in prostate cancer is step by step being elucidated. These recent advances are here described and discussed. PMID- 21623388 TI - Prostate cancer cells metastasize to the hematopoietic stem cell niche in bone. PMID- 21623389 TI - Efficacy and cost analysis of transrectal ultrasound-guided prostate biopsy under monitored anesthesia. AB - Sedation may result in reduction in pain during transrectal ultrasound (TRUS) guided prostate biopsies. We aimed to evaluate the efficacy and safety of a combination of propofol and remifentanil infusion during TRUS-guided prostate biopsy and the related increases in health care costs. From January to September 2010, 100 men undergoing a transrectal prostate biopsy were randomized into two groups. In Group 1, 50 patients received a combined infusion of propofol and remifentanil; in Group 2, 50 patients received lidocaine jelly. After TRUS-guided biopsies were performed, pain and patient satisfaction were evaluated by a 10 point visual analog scale (VAS), and a cost-related patient satisfaction questionnaire was completed by all patients. Patients were also asked whether they would be willing to undergo repeat biopsy by the same method. Patients in Group 1 showed a significantly lower VAS score than those in Group 2 (mean VAS score: 0.9+/-1.1 versus 6.3+/-2.5; P<0.001). In addition, the patient satisfaction scale was significantly higher in Group 1 (P=0.002). Although the overall cost was significantly higher in Group 1 (P=0.006), patient satisfaction scales considering cost were also higher in this group (P=0.009). A combination of propofol and remifentanil is a safe and effective way to decrease patient pain and increase patient satisfaction during TRUS-guided prostate biopsy. Although the costs were higher in the group that received sedation, as expected, the patients exhibited heightened satisfaction and willingness to repeat biopsies by the same method. PMID- 21623390 TI - Activation of human ether-a-go-go related gene (hERG) potassium channels by small molecules. AB - Human ether-a-go-go related gene (hERG) potassium (K(+)) channels play a critical role in cardiac action potential repolarization. Mutations that reduce hERG conductance or surface expression may cause congenital long QT syndrome (LQTS). However, the channels can be inhibited by structurally diverse small molecules, resulting in an acquired form of LQTS. Consequently, small molecules that increase the hERG current may be of value for treatment for LQTS. So far, nine hERG activators have been reported. The aim of this review is to discuss recent advances concerning the identification and action mechanism of hERG activators. PMID- 21623391 TI - Role of the epithelial sodium channel in salt-sensitive hypertension. AB - The epithelial sodium channel (ENaC) is a heteromeric channel composed of three similar but distinct subunits, alpha, beta and gamma. This channel is an end effector in the rennin-angiotensin-aldosterone system and resides in the apical plasma membrane of the renal cortical collecting ducts, where reabsorption of Na(+) through ENaC is the final renal adjustment step for Na(+) balance. Because of its regulation and function, the ENaC plays a critical role in modulating the homeostasis of Na(+) and thus chronic blood pressure. The development of most forms of hypertension requires an increase in Na(+) and water retention. The role of ENaC in developing high blood pressure is exemplified in the gain-of-function mutations in ENaC that cause Liddle's syndrome, a severe but rare form of inheritable hypertension. The evidence obtained from studies using animal models and in human patients indicates that improper Na(+) retention by the kidney elevates blood pressure and induces salt-sensitive hypertension. PMID- 21623393 TI - Obesity and chronic kidney disease. AB - Obesity is associated with the early onset of glomerulomegaly, hemodynamic changes of a hyperfiltering kidney, and increased albuminuria, which are potentially reversible with weight loss. However, pathologic lesions of focal segmental glomerulosclerosis develop in experimental models of sustained obesity, and are observed in morbidly obese humans presenting with massive proteinuria. In addition, several observational, cross sectional and longitudinal studies document that obesity is as an independent risk factor for the onset, aggravated course, and poor outcomes of chronic kidney disease, even after adjustment for confounding co-morbidities including metabolic syndrome, diabetes and hypertension, the major causes of chronic kidney disease. Early dietary intervention to reduce weight, and where necessary bariatric surgery, should be considered in the management of overweight and obese chronic kidney disease (CKD) patients. PMID- 21623392 TI - Nanomechanical analysis of insulinoma cells after glucose and capsaicin stimulation using atomic force microscopy. AB - AIM: Glucose stimulates insulin secretion from pancreatic islet beta cells by altering ion channel activity and membrane potential in the beta cells. TRPV1 channel is expressed in the beta cells and capsaicin induces insulin secretion similarly to glucose. This study aims to investigate the biophysical properties of the beta cells upon stimulation of membrane channels using an atomic force microscopic (AFM) nanoindentation system. METHODS: ATCC insulinoma cell line was used. Cell stiffness, a marker of reorganization of cell membrane and cytoskeleton due to ion channel activation, was measured in real time using an integrated AFM nanoindentation system. Cell height that represented structural changes was simultaneously recorded along with cell stiffness. RESULTS: After administration of glucose (16, 20 and 40 mmol/L), the cell stiffness was markedly increased in a dose-dependent manner, whereas cell height was changed in an opposite way. Lower concentrations of capsaicin (1.67 * 10(-9) and 1.67 * 10(-8) mol/L) increased the cell stiffness without altering cell height. In contrast, higher concentrations of capsaicin (1.67 * 10(-6) and 1.67 * 10(-7) mol/L) had no effect on the cell physical properties. CONCLUSION: A unique bio-nanomechanical signature was identified for characterizing biophysical properties of insulinoma cells upon general or specific activation of membrane channels. This study may deepen our understanding of stimulus-secretion coupling of pancreatic islet cells that leads to insulin secretion. PMID- 21623394 TI - Surveillance for infections and other adverse events in dialysis patients in southern Gran Canaria. AB - BACKGROUND: Bacterial infections pose a major challenge to risk management activities in the area of chronic haemodialysis, as vascular access-related infections are the main cause of mortality among these patients. METHODS: Prospective surveillance study lasting 7 months (March-September, 2008) at two haemodialysis units in a district health area Gran Canaria, Spain. We used the methodology proposed by CDC's Dialysis Surveillance Network. RESULTS: 1545 patients/month were recorded, 60.5% with an arteriovenous fistula (AVF), 35.5% with a permanent catheter (PC), 3.0% with grafts and 1.0% with temporary catheters. The rate of adverse events was 8.6 cases per 100 patients/month, 9.1 for AVF patients, and 2.9 for PC. Nevertheless, the other types of infections (respiratory, urinary tract, skin and chronic ulcers) showed similar rates. Microbiological cultures were taken in 82.2%, but this rate increased to 91.0% when a vascular access-related infection was suspected. Empirical treatment was adjusted to antibiogram results in 90.0% of occasions. A low incidence of multi resistant microbes was observed. Gram-positive and gram-negative bacteria appeared in similar proportions. CONCLUSIONS: Vascular access is the main risk factor for infectious events. Epidemiological surveillance has allowed us to detect areas of improvement in different settings, acting as a key element in risk management and patient safety. PMID- 21623395 TI - The Cerebral Circulation: Some New Points in its Anatomy, Physiology and Pathology. PMID- 21623396 TI - An Investigation of the Absorption of Ultraviolet Light by Cerebrospinal Fluid in Various Disease States. PMID- 21623397 TI - Associated Facial, Vocal and Respiratory Components of Emotional Expression: An Experimental Study. PMID- 21623398 TI - Paroxysmal Trigeminal Pain with Tumours of the Nervus Acusticus. PMID- 21623399 TI - The Diagnosis of Chronic Subdural HaeMatoma of Traumatic Origin. PMID- 21623400 TI - A Gap in Neuropsychiatric Knowledge. PMID- 21623402 TI - Heme Bodies (Rosenthal Fibres) associated with Cavities in Pons and Cerebellum and Acoustic Neurinoma: With a Report of Two Cases. PMID- 21623401 TI - An Enquiry into the Causes of Mescal Visions. PMID- 21623403 TI - The Blood Calcium In 'Idiopathic' Epilepsy. PMID- 21623405 TI - Tissue Culture Methods in the Study of the Nervous System: A Review. PMID- 21623404 TI - Loneliness and the Paranoid Syndrome. PMID- 21623406 TI - Amaurotic Idiocy and the Lipoidoses. PMID- 21623407 TI - TRAUMATIC DILATATION OF THE CEREBRAL VENTRICLES. PMID- 21623408 TI - THE EFFECT OF GALVANIC EXERCISE ON DENERVATED AND RE-INNERVATED MUSCLES IN THE RABBIT. PMID- 21623409 TI - MYELITIS DUE TO VACCINATION. PMID- 21623410 TI - THE PHYSICAL EXAMINATION OF TWO THOUSAND CASES OF NEUROSIS. PMID- 21623411 TI - CAUSATION OF MONGOLISM. PMID- 21623412 TI - A CASE OF PARTIAL CONGENITAL HEMI-HYPERTROPHY. PMID- 21623413 TI - POST-TRAUMATIC PAIN AND THE CAUSALGIC SYNDROME. PMID- 21623414 TI - A HEURISTIC THEORY OF NEUROSIS. PMID- 21623415 TI - Phosphorylation of basic amino acid residues in proteins: important but easily missed. AB - Reversible phosphorylation is the most widespread posttranslational protein modification, playing regulatory role in almost every aspect of cell life. The majority of protein phosphorylation research has been focused on serine, threonine and tyrosine that form acid-stable phosphomonoesters. However, protein histidine, arginine and lysine residues also may undergo phosphorylation to yield acid-labile phosphoramidates, most often remaining undetected in conventional studies of protein phosphorylation. It has become increasingly evident that acid labile protein phosphorylations play important roles in signal transduction and other regulatory processes. Beside acting as high-energy intermediates in the transfer of the phosphoryl group from donor to acceptor molecules, phosphohistidines have been found so far in histone H4, heterotrimeric G proteins, ion channel KCa3.1, annexin 1, P-selectin and myelin basic protein, as well as in recombinant thymidylate synthase expressed in bacterial cells. Phosphoarginines occur in histone H3, myelin basic protein and capsidic protein VP12 of granulosis virus, whereas phospholysine in histone H1. This overview of the current knowledge on phosphorylation of protein basic amino-acid residues takes into consideration its proved or possible roles in cell functioning. Specific requirements of studies on acid-labile protein phosphorylation are also indicated. PMID- 21623416 TI - Metabolism of bradykinin in aorta of hypertensive rats. AB - Alterations in the formation and metabolism of bradykinin (Bk) are hypothesized to play a role in the pathophysiology of hypertension, atherosclerosis and vascular complications of diabetes. However, despite its prominent role in cardiovascular regulation, studies on bradykinin have been limited by various difficulties in accurate measurements of this peptide in biological samples. In this study, using the LC-ESI-MS method we estimated the conversion of exogenous Bk to its main metabolites - Bk-(1-5) and Bk-(1-7) - in endothelial cell culture and in fragments of aorta of normotensive (WKY) and hypertensive rats (SHR). The effects of angiotensin converting enzyme (ACE) and neutral endopeptidase (NEP) inhibitors were more pronounced in SHR: perindoprilat inhibited Bk-(1-5) formation by 49 % and 76 % in WKY and SHR rats, respectively, and tiorphan tended to decrease formation of Bk-(1-5) in both groups of animals. The degradation of bradykinin and generation of both metabolites were significantly higher in the aorta of SHR rats than in WKY controls. Our results show that even in relatively early hypertension (in 4-month old SHR rats) inactivation of Bk by aorta wall is enhanced. PMID- 21623418 TI - Remodeling and repair of orthopedic tissue: role of mechanical loading and biologics. AB - Orthopedic tissues respond to mechanical loads to maintain normal homeostasis and in response to injury. As the body of work on this continues to grow, it is important to synthesize the recent studies across tissues and specialties with one another and with past studies. Hence, this review highlights the knowledge gained since 2000, with only few exceptions, concerning the effects of mechanical load and biologics on remodeling and repair of orthopedic tissue. PMID- 21623419 TI - Augmented demineralized bone matrix: a potential alternative for posterolateral lumbar spinal fusion. AB - Variable osteoinductive potential has been reported between and within production lots of different demineralized bone matrix (DBM) products. This study compared fusion rates of different manufactured lots and augmented formulations of DBM with a dose-response curve of recombinant human bone morphogenetic protein 2 (rhBMP-2) on inactivated DBM carrier in a posterolateral fusion rat model. Lumbar fusions were performed in 145 rats. In the control rats, we implanted autograft, graft alternative, including inactivated DBM, or nothing (ie, no graft). In the study rats, we implanted 1 of 2 BioSETR (RTI Biologics, Alachua, Florida) DBM lots, growth factor-enriched DBM, and inactivated DBM plus rhBMP-2 in different concentrations. Manual palpation revealed fusion rates of 25% (autograft), 0% (inactivated DBM), 17% (DBM donor A), and 36% (DBM donor B). The fusion rate of the most enhanced donor B graft (83%) was higher (P<.05) than that of autograft or unenhanced DBM. Inactivated DBM plus rhBMP-2 fused between 45% and 100%. There was no significant difference between DBM plus rhBPM-2 and the highest enrichment group of donor B. Differences between 2 DBM lots in an athymic rat ectopic bone formation model also were found in the spine fusion model. Enhanced DBM formulations were comparable with inactivated DBM plus rhBMP-2 with respect to performance and could represent a bone graft alternative in spine fusion. PMID- 21623420 TI - Revision extensor mechanism allografting after total knee arthroplasty. AB - This study analyzed the outcomes of revision extensor mechanism allografting (EMA) in 8 patients with failures of previous EMA after total knee arthroplasty. Fresh-frozen allografts were used. Knee Society Clinical Rating System (KSS) scores were used to measure clinical outcomes, with special attention given to presence of extensor lag and assistive ambulatory devices. Patients were observed for an average of 3.1 years. Two patients were considered failures due to infection within the first postoperative year of the revision EMA. In the remaining 6 patients, the average KSS score at most recent follow-up showed no improvement when compared with preoperative scores. Revisions of the failed EMA resulted in a high incidence of functional limitations, which continue to deteriorate over time. PMID- 21623421 TI - Proximal rectus femoris avulsion in an elite, olympic-level sprinter. AB - Quadriceps injuries, ranging from simple strains to disabling muscle ruptures, are common athletic injuries. The rectus femoris is the most commonly injure portion of the quadriceps musculature. This article is, to our knowledge, the first report of a proximal rectus femoris avulsion in an elite, Olympic-level 100 meter sprinter, acutely managed with surgical repair. Several key factors must be considered and carefully assessed when determining the appropriate course of management (ie, deciding between operative and nonoperative treatment): amount of distal retraction of the tendon, severity of associated soft-tissue trauma, physical examination, and postoperative goals (eg, return to elite-level competitive sports involving running or kicking vs resuming basic activities of daily living). We believe that these factors in our elite, high-performance athlete dictated an operative course of management. PMID- 21623422 TI - Hemophilic arthropathy. PMID- 21623423 TI - Dedifferentiated liposarcoma of thigh with chondrosarcomatous dedifferentiated component. AB - Liposarcomas are common soft-issue sarcomas arising predominantly in deep soft tissue and the retroperitoneum with varied mortality and recurrence rates, largely dependent on histologic type. Thought to arise de novo, liposarcomas typically are classified into 5 types based on strict morphologic characteristics: well-differentiated, dedifferentiated, myxoid, round cell, and pleomorphic. More specifically, dedifferentiated liposarcoma, a common type most prevalent in the retroperitoneum, often has 2 distinct components, a well differentiated lipomatous component and a dedifferentiated nonlipomatous component composed of sarcomas, such as myxofibrosarcomas or other spindle-cell sarcomas. Morphology typically ranges from low- to high-grade components, most commonly exhibiting myxofibrosarcoma and malignant fibrous histiocytoma components. However, the case reported in this article is unique-the dedifferentiated component exhibited only chondrosarcomatous differentiation-and it is, to our knowledge, the first such case to be described. PMID- 21623424 TI - Recurrent knee pain in an athletic adult: multiple schwannomas secondary to schwannomatosis: a case report. AB - Schwannomatosis has been used to describe patients with multiple nonvestibular schwannomas with no associated features of neurofibromatosis type 2. In our case, a 28-year-old athletic man underwent a right knee excisional biopsy for multifocal, benign schwannomatosis. After being asymptomatic for 4 years postresection, he returned to our musculoskeletal oncology service. Imaging studies revealed local recurrence identical to his initial presentation. Excisional biopsy of discrete masses was performed and histologic examination revealed recurrent benign schwannomatosis. To our knowledge, this is the second reported case of recurrent benign schwannomatosis. We review schwannomatosis, including its etiology, radiographic features, and relationship to neurofibromatosis. PMID- 21623425 TI - Extranodal rosai-dorfman disease presenting as incidental bone tumor: a case report. AB - We report a case of primary extranodal Rosai-Dorfman disease presenting as a painless lesion in the left ilium of a 71-year-old African-American man. PMID- 21623426 TI - A highly active and selective palladium pincer catalyst for the formation of alpha-aryl ketones via cross-coupling. AB - Several air and moisture stable Pd(II) pincer complexes were synthesized via oxidative addition of Pd(0) to novel PheBox pincer ligand precursors. Low loadings (1 mol%) of the Pd complex [t-BuPhebox-Me(2)]PdBr are capable of efficiently promoting the selective alpha-monoarylation of a variety of ketones with numerous aryl bromides in only 1 h at 70 degrees C with 82-99% yields. PMID- 21623427 TI - Molybdenum metabolism in the alga Chlamydomonas stands at the crossroad of those in Arabidopsis and humans. AB - Molybdenum (Mo) is a very scarce element whose function is fundamental in living beings within the active site of Mo-oxidoreductases, playing key roles in the metabolism of N, S, purines, hormone biosynthesis, transformation of drugs and xenobiotics, etc. In eukaryotes, each step from Mo acquisition until its incorporation into a biologically active molybdenum cofactor (Moco) together with the assembly of this Moco in Mo-enzymes is almost understood. The deficiency in function of a particular molybdoenzyme can be critical for the survival of the organism dependent on the pathway involved. However, incapacity in forming a functional Moco has a pleiotropic effect in the different processes involving this cofactor. A detailed overview of Mo metabolism: (a) specific transporters for molybdate, (b) the universal biosynthesis pathway for Moco from GTP, (c) Moco carrier and Moco-binding proteins for Moco transfer and (d) Mo-enzymes, is analyzed in light of recent findings and three systems are compared, the unicellular microalga Chlamydomonas, the plant Arabidopsis and humans. PMID- 21623428 TI - Boosting electrical conductivity in a gel-derived material by nanostructuring with trace carbon nanotubes. AB - An organogelator with two distinct pi-functional units is able to incorporate carbon nanotubes into its mesh of fibres in the gel state. The morphology of the material derived from this nanocomposite after evaporation of the solvent is a complex mesh of fibres which is clearly different from the pure gelator. This feature indicates a role of the nanotubes in assisting the formation of a fibre structure in the gel thanks to their interaction with the pyrene units in the organogelator. The nanocomposite conducts electricity once the p-type gelator is doped with iodine vapour. The change in morphology caused by the carbon material increases the conductivity of the material compared with the purely organic conducting system. It is remarkable that this improvement in the physical property is caused by an extremely small proportion of the carbon material (only present at a ratio of 0.1% w/w). The practically unique properties of TTF unit allow measurements with both doped and undoped materials with conducting atomic force microscopy which have demonstrated that the carbon nanotubes are not directly responsible for the increased conductivity. PMID- 21623429 TI - 9G DNAChip: microarray based on the multiple interactions of 9 consecutive guanines. AB - We introduce the phenomenon of molecular recognition to immobilize oligonucleotides on AMCA slides for the production of 9G DNAChips. Facile and efficient method for the immobilization of the oligonucleotides appended with consecutive nine guanine bases is described. The 9G DNAChips shows more than 90% hybridization efficiency at 25 degrees C in 30 min. PMID- 21623430 TI - Study of structural and dynamic properties of liquid phenyltrimethoxysilane. AB - Herein, we present a combined experimental and computational study of liquid phenyltrimethoxysilane. A femtosecond time-resolved optical Kerr effect experiment has been performed to study the rotational diffusion of the molecule. A new all-atoms molecular model of the compound, based on the OPLS force field, has been developed to reproduce the rotational diffusion time constant and other physical and dynamic properties available in the literature. The density obtained from the simulations is 1074 +/- 4 kg m(-3), which is within 1% of the experimental value of 1062 kg m(-3). The viscosity from the simulations is 1.6 +/ 0.1 mPa s while the experimental value is 2.1 mPa s. The average bulk dipole moment of 1.8 +/- 0.5 Debye obtained from the simulation matches the experimental value of 1.77 Debye. The average relative dielectric constant from the simulations is 3.86 +/- 0.04, which is within 13% of the experimental value (4.4). The rotational diffusion time of the dipole moment obtained from the simulations is 20.39 +/- 0.06 ps, which is in excellent agreement with the experimental value of 20 +/- 1 ps obtained from our measurements. The new model has also been used to calculate structural and dynamic properties of the molecule not yet determined experimentally. PMID- 21623431 TI - Combined effect of stacking and solvation on the spontaneous mutation in DNA. AB - In DNA, base pairs are involved in two reciprocal interactions: interbase hydrogen bonds and stacking. Furthermore, base pairs also undergo the effects of the external entities present in the biological environment, such as water molecules and cations. In this contribution, the double spontaneous mutation has been studied with hybrid theoretical tools in a DNA-embedded guanine-cytosine model accounting for the impact of the first hydration shell. According to our findings, the combination of the neighboring base pairs and surrounding water molecules plays a crucial role in the double proton transfer. Indeed, as a consequence of these interactions, the double proton transfer (DPT) mechanism is altered: on the one hand, stacking and hydration strongly affect the geometry of base pairs, and, on the other hand, vicinal water molecules may play an active role in the tautomeric equilibrium by catalyzing the proton transfer reaction. PMID- 21623432 TI - Monitoring the status of T-cell activation in a microfluidic system. AB - Leukocyte adhesion to the endothelium through surface molecules such as E selectin and intercellular adhesion molecule-1 (ICAM-1) is a critical cellular event reflecting the physiological status of both cell types. Here we present a microfluidic system that can not only easily monitor the interaction between leukocytes and endothelial cells under physiological conditions, but also screen drug candidates for potential modulation of this interaction. Shear stress, which is an important factor for the binding of activated T cells to tumor necrosis factor-alpha (TNF-alpha)-treated human umbilical vein endothelial cells (HUVECs), was easily controlled by adjusting the flow rate in the microfluidic system. Whole blood of patients with systemic lupus erythematosus (SLE) who have auto reactive T cells were infused into the activated HUVECs which subsequently showed a higher level of binding compared to a control blood sample from a person without SLE. When these autoreactive T cells were treated with immunosuppressors tacrolimus and cyclosporin A, the binding of the T cells to HUVECs was dramatically decreased. Therefore, this microfluidic system is capable of differentiating the physiological status of T cells or endothelial cells representing different disease conditions, as well as being useful for the identification of novel reagents that modulate the functions of leukocytes or endothelial cells. PMID- 21623433 TI - Arylation of alpha-pivaloxyl ketones with arylboronic reagents via Ni-catalyzed sp3 C-O activation. AB - A Suzuki-Miyaura coupling of alpha-pivaloxyl ketones via Ni-catalyzed sp(3) C-O activation to produce alpha-aryl ketones is developed. This study offers a convenient method to construct alpha-arylation products from readily available alpha-hydroxyl carbonyl compounds. PMID- 21623434 TI - A highly selective colorimetric and ratiometric fluorescent chemodosimeter for imaging fluoride ions in living cells. AB - A simple but highly selective colorimetric and ratiometric fluorescent chemodosimeter was designed and synthesized to detect fluoride ions (F(-)) in aqueous solution and living cells by virtue of the strong affinity of F(-) toward silicon. PMID- 21623435 TI - A salt made of 4-N,N-dimethylaminopyridine (DMAP) and saccharin as an efficient recyclable acylation catalyst: a new bridge between heterogeneous and homogeneous catalysis. AB - Here, we report insights into the new recyclable catalyst 1, (DMAP.saccharin). The DMAP.saccharin-catalysed acylation of alcohols has been successfully carried out more than 8 times. Only 1 mol% of catalyst 1 efficiently promotes acylation with almost equimolar amounts of acid anhydrides, under both base-free and solvent-free conditions. PMID- 21623436 TI - Efficient bifunctional gallium-68 chelators for positron emission tomography: tris(hydroxypyridinone) ligands. AB - A new tripodal tris(hydroxypyridinone) bifunctional chelator for gallium allows easy production of (68)Ga-labelled proteins rapidly under mild conditions in high yields at exceptionally high specific activity and low concentration. PMID- 21623437 TI - Supported lipid bilayer microarrays created by non-contact printing. AB - Arrays of supported lipid bilayers (SLBs) provide great potential for future drug development and multiplexed biological research, but are difficult to prepare due to the sensitivity of both the lipid and protein structural arrangement to air exposure. A novel way to produce arrays of SLBs is presented based on non-contact dispensing of vesicles to a substrate through a thin surface confined water film. The approach presents many degrees of freedom since it is not limited to a specific substrate, lipid composition, linker or controlled environment. The method allows adjustment of spot size (180-360 MUm) by repeated dispensing as well as control over the composition of the spots and subsequent analytes. SLB formation by vesicle adsorption and rupture allows for incorporation of membrane proteins through pre-formed proteoliposomes. Dispensing through a dip-and-rinse water film avoids contamination, disruptive drying and the need for complex buffer compositions. Furthermore, no humidity control is necessary which simplifies the production step and prolongs the life-time of the spotting system. We characterize the method with respect to control over spot size, bilayer mobility and the formation process as well as demonstrate the possibility to fuse bilayer spots with subsequently added vesicles. Since complex lipid compositions and multiple spotting nozzles can be used, this novel technique is expected to be a promising platform for future applications, e.g. patterning to monitor peptide/protein-lipid interactions, for glycomics using glycolipids or lipopolysaccharides, and to study mixing of spatially confined lipid membranes. PMID- 21623438 TI - High throughput label-free platform for statistical bio-molecular sensing. AB - Sensors are crucial in many daily operations including security, environmental control, human diagnostics and patient monitoring. Screening and online monitoring require reliable and high-throughput sensing. We report on the demonstration of a high-throughput label-free sensor platform utilizing cantilever based sensors. These sensors have often been acclaimed to facilitate highly parallelized operation. Unfortunately, so far no concept has been presented which offers large datasets as well as easy liquid sample handling. We use optics and mechanics from a DVD player to handle liquid samples and to read out cantilever deflection and resonant frequency. Also, surface roughness is measured. When combined with cantilever deflection the roughness is discovered to hold valuable additional information on specific and unspecific binding events. In a few minutes, 30 liquid samples can be analyzed in parallel, each by 24 cantilever-based sensors. The approach was used to detect the binding of streptavidin and antibodies. PMID- 21623439 TI - One-pot synthesis of sustained-released doxorubicin silica nanoparticles for aptamer targeted delivery to tumor cells. AB - Site-specific delivery of drugs can significantly reduce drug toxicity and increase the therapeutic effect. Here, we report a one-pot synthesis of doxorubicin-doped silica nanoparticles (Dox/SiNPs) by using sodium fluoride (NaF) catalyzed hydrolysis of tetraethyl orthosilicate in a water-in-oil microemulsion. Through further surface chemical modification, carboxyl-terminated Dox/SiNPs (COOH-Dox/SiNPs) exhibiting high drug entrapment efficiency, strong fluorescence and long sustained release are obtained. Cell toxicity tests demonstrate that the COOH-Dox/SiNPs kill tumor cells effectively, while pure COOH-SiNPs are nontoxic. An aptamer is further conjugated to the nanoparticles for delivering loaded Dox to target cells. It is demonstrated that Dox/SiNPs modified with the aptamer sgc8c (sgc8c-Dox/SiNPs) could deliver loaded doxorubicin to CCRF-CEM cells with high specificity and excellent efficiency. Furthermore, ex vivo imaging studies show that the COOH-Dox/SiNPs are able to accumulate highly in the tumor areas, thanks to the enhanced permeability and retention (EPR) effects. Our data suggest that the sgc8c-Dox/SiNPs may be a useful new tumor therapy system. PMID- 21623440 TI - Heat and light dual switching of a single-walled carbon nanotube/thermo responsive helical polysaccharide complex: a new responsive system applicable to photodynamic therapy. AB - A thermo- and light-responsive system consisting of single-walled carbon nanotube and helical polysaccharide modified with poly(N-isopropylacrylamide) side-chains has been developed through supramolecular polymer wrapping. Coagulation of the complex can be induced by the external stimuli, which leads to a catch-and release action of a porphyrin derivative. PMID- 21623441 TI - Photoluminescent selenospirocyclic and selenotetracyclic derivatives by domino reactions of amines and imines. AB - Novel photoluminescent selenospirocyclic and selenotetracyclic compounds, stabilized by intramolecular secondary Se...O interactions with an ortho-nitro group, have been accessed successfully through domino reactions of amines and imines, respectively. PMID- 21623442 TI - PNA-based artificial nucleases as antisense and anti-miRNA oligonucleotide agents. AB - Because of its interesting chemical, physical and biological properties, Peptide Nucleic Acid (PNA) has attracted major attention in molecular biology, for diagnostics purposes and development of biosensors. PNAs have become candidates for gene therapeutic drugs in ANTISENSE (AO) strategy with favorable in vivo biochemical properties. Recently, antisense PNA oligonucleotides have been described in anti-miRNA approach (AMO). We propose PNA-based nucleases as AO and AMO agents. We report the design, synthesis and characterization of two kinds of artificial nucleases composed of a PEG-PNA-PEG domain conjugated to HGG.Cu (A) and DETA (B) as well known cleavage sites. Qualitative (MALDI-TOF) and quantitative (HTS) assays were planned to study nuclease activity of constructs A and B on RNA-3'-FAM target sequence. The results have highlighted the best performance of nuclease B and the relevance of the PEG spacer, in particular for conjugate A, in terms of efficiency of the cleavage, suggesting that conjugates A and B also act as potential antisense and anti-miRNA agents. PMID- 21623443 TI - Towards experimental determination of conical intersection properties: a twin state based comparison with bound excited states. AB - The energy and approximate structure of certain S(0)/S(1) conical intersections (CI) are shown computationally to be deducible from those of two bound states: the first triplet (T(1)), which is iso-energetic with the CI, and the second excited singlet state (S(2)). This is demonstrated for acepentalene (I) and its perfluoro derivative (II) using the twin state concept for three states systems and based on the fact that the triplet T(1) is almost degenerate with the CI. The stable S(2) (C(3v) configuration) state exhibits unusual exaltation of Jahn Teller active degenerate mode-nu(JT) = 2058 cm(-1) (~500 cm(-1) higher than analogous e-mode of the symmetric (C(3v)) T(1) and the dianion I(-2) or any C-C vibration of the Jahn-Teller distorted (C(s)) ground state minimum). The acepentalene molecule, whose rigid structure and possibility to attain the relatively high symmetry C(3v) configuration, is a particularly suitable candidate for this purpose. PMID- 21623448 TI - Vibrational predissociation dynamics of Cl2(B)-He2: a wave packet study. AB - The vibrational predissociation of Cl(2)(B)-He(2) has been studied using a full dimensional wave packet method. The aim is to investigate the effect of increasing the grid size in the dissociative coordinates and the propagation time, on the convergence of observable magnitudes like predissociation lifetimes and Cl(2) product vibrational and rotational distributions. In particular, convergence of vibrational distributions is significantly affected by an artifact caused by the use of finite grids and absorbing conditions for the wave packet, combined with the presence of a sequential dissociation process. The results show that the lifetimes and the Cl(2)(B) rotational distributions are not greatly affected by increasing propagation time and grid size. However, convergence of the Cl(2) vibrational distribution is very slow, and the strategy of converging this property by increasing the grid size becomes impractical. An approximate model to estimate the Cl(2) vibrational populations is suggested, which is found to provide realistic distributions as compared with the available experimental ones. The main feature of the model is that its assumptions are closely based on the nature of the vibrational predissociation process occurring in the type of complexes. This feature of the model, in addition to its simplicity of implementation and negligible extra computational cost, contributes to the general applicability of the approach to BC(B)-Rg(2) complexes. PMID- 21623449 TI - Emission-photoactivity cross-processing of mesoporous interfacial charge transfer in Eu3+ doped titania. AB - Periodic mesoporous Eu(3+) doped titania materials were obtained through the EISA (Evaporation Induced Self Assembly) process. Eu(3+) ions, entrapped within the semi-crystalline walls of the highly porous framework, appear to be advantageous during the probing of surface photochemical reactions. Its emission intensity is very sensitive to the presence of physisorbed molecules, in gas or liquid phase, that reside within the pores. In particular, strong fluctuations in intensity of the (5)D(0)->(7)F(2) transition were observed under UV light exposure on the time scale of tens of seconds. The emission modulation dynamics show a strong correlation with the crystallinity of the titania matrix. Correlation of the emission with the photocatalytic activity of the semiconductor for photodegradation of an organic molecule is observed. A model is proposed to describe the involved mechanisms. PMID- 21623450 TI - Influence of surface chemistry and protein concentration on the adsorption rate and S-layer crystal formation. AB - Bacterial crystalline surface layers (S-layers) are the outermost envelope of prokaryotic organisms representing the simplest biological membranes developed during evolution. In this context, the bacterial protein SbpA has already shown its intrinsic ability to reassemble on different substrates forming protein crystals of square lattice symmetry. In this work, we present the interaction between the bacterial protein SbpA and five self-assembled monolayers carrying methyl (CH(3)), hydroxyl (OH), carboxylic acid (COOH) and mannose (C(6)H(12)O(6)) as functional groups. Protein adsorption and S-layer formation have been characterized by atomic force microscopy (AFM) while protein adsorption kinetics, mass uptake and the protein layer viscoelastic properties were investigated with quartz crystal microbalance with dissipation monitoring (QCM-D). The results indicate that the protein adsorption rate and crystalline domain area depend on surface chemistry and protein concentration. Furthermore, electrostatic interactions tune different protein rate adsorption and S-layer recrystallization pathways. Electrostatic interactions induce faster adsorption rate than hydrophobic or hydrophilic interactions. Finally, the shear modulus and the viscosity of the recrystallized S-layer on CH(3)C(6)S, CH(3)C(11)S and COOHC(11)S substrates were calculated from QCM-D measurements. Protein-protein interactions seem to play a main role in the mechanical stability of the formed protein (crystal) bilayer. PMID- 21623453 TI - Emission rate of particulate matter and its removal efficiency by precipitators in under-fired charbroiling restaurants. AB - In order to explore the potent role of meat cooking processes as the emission sources of particulate matter (PM), emission rates and the associated removal efficiency by precipitators were estimated based on the on-site measurements made at five under-fired charbroiling (UFC) restaurants. The emission patterns of PM for these five restaurants were compared after having been sorted into the main meat types used for cooking: beef (B), chicken (C), intestines (I), and pork (P: two sites). The mass concentrations (microgram m-3) of three PM fractions (PM2.5/PM10/TSP) measured from these restaurants were 15,510/15,701/17,175 (C); 8,525/10,760/12,676 (B); 11,027/13,249/13,488 (P); and 22,409/22,412/22,414 (I). Emission factors (g kg-1) for those PM fractions were also estimated as 3.23/4.08/4.80 (B), 3.07/3.82/3.87 (P), 8.12/8.22/8.99 (C), and 6.59/6.59/6.59 (I). If the annual emission rate of PM10 is extrapolated by combining its emission factor, population, activity factor, etc., it is estimated as 500 ton year-1, which corresponds to 2.4% of the PM10 budget in Seoul, Korea. Removal efficiencies of PM10 via precipitators, such as an electrostatic precipitator (ESP), bag filter (BF), and the combination system (ESP + catalyst), installed in those UFC restaurants ranged between 54.76 and 98.98%. The removal efficiency of PM by this control system was the least effective for particles with <0.4 microgram, although those in the range of 0.4-10 microgram were the most effective. PMID- 21623454 TI - Effectiveness of a positive youth development program for secondary 1 students in Macau: a pilot study. AB - With the rapid change to society after the opening of the gaming licensure by the government and the potential attraction to youth caused by the casinos, a well tested and comprehensive adolescent development program previously established in Hong Kong was adopted and modified to be used in Macau. It is expected to help our adolescents achieve positive growth and be better prepared for future challenges. The aim of this study is to examine the effectiveness of the modified positive youth development program for Secondary 1 Students in Macau. Specifically, two research questions will be asked: (1) How does the positive youth development program affect positive growth for youth in Macau?; and (2) Is youth growth related to different factors such as gender, age, family financial condition, and parents' marital status? A mixed research method with a quantitative approach using a pre- and post-test pre-experimental design, and a qualitative approach using a focus group for the participants is carried out. The study sample included 232 Secondary 1 Students in two schools. The objective outcome evaluation showed that, overall, 123 (53%) of the participants had significant improvement on the total scores of the Chinese Positive Youth Development Scale (CPYDS) and the two composite scores. However, there were some increases in the behavioral intention of alcohol drinking and participation in gambling activities. The "happiness of the family life" was found to have significant differences in the score of the CPYDS, which was shown to be the factor related to youth growth. The focus group interviews revealed that both positive and negative feedback was obtained from the discussion; however, the majority of the participants perceived benefits to themselves from the program. With reference to the principle of triangulation, the present study suggests that, based on both quantitative and qualitative evaluation findings, it should be concluded that there is positive evidence supporting the effectiveness of the Tier 1 Program of the Hong Kong Project P.A.T.H.S. (Positive Adolescent Training through Holistic Social Programmes), which was adopted and modified for Macau. In addition, special attention should be paid to the behavioral intention of alcohol drinking and participation in gambling activities in the local context. PMID- 21623455 TI - Green urine. AB - Urinalysis is an integral part of a thorough patient evaluation. Change in urine characteristics can give clues to help solve some of the diagnostic challenges faced by physicians. We discuss a case of a benign cause of green discoloration of urine caused by propofol infusion, which reversed following its discontinuation. PMID- 21623456 TI - Current and emerging bladder cancer urinary biomarkers. AB - Bladder cancer continues to be one of the most common malignancies. Those who have been already diagnosed are at high risk for recurrence, especially if the pathology demonstrates high-grade disease. Diagnosis and surveillance is reliant on invasive evaluation with cystoscopy. Urinary cytology has been used to aid in diagnosis, but its use is limited. Other assays have been developed that may aid in clinical decision making. The ultimate goal will be the development of a highly sensitive and specific urinary marker for bladder cancer. This would provide a noninvasive means of diagnosing the disease and limit the number of unnecessary cystoscopies. This article will review the currently available urinary bladder cancer markers. It will also review new and investigational urinary markers that have shown promise for future clinical use. PMID- 21623457 TI - Synthesis and Anti-Mycobacterium tuberculosis Evaluation of Aza-Stilbene Derivatives. AB - Tuberculosis (TB) is a truly global disease, found in every country on earth. One third of humanity, over 2 billion people, carry the bacillus that causes TB and 2 million people die of the disease each year. Despite that, no new specific drug against Mycobacterium tuberculosis has been developed since the 1960s. There are several candidates for new anti-TB agents, but none proven clinically effective. Stilbenes are compounds found in numerous medicinal plants and food products with some known biological and even antimycobacterial activity. This paper describes the synthesis and the anti-M. tuberculosis activity of eight stilbene analogues. The synthesis and characterization of these compounds are shown, and the results compared with one "first"-line drug used in current therapy. PMID- 21623458 TI - Extracellular vesicles derived from Acholeplasma laidlawii PG8. AB - Extracellular vesicle production is believed to be a ubiquitous process in bacteria, but the data on such a process in Mollicutes are absent. We report the isolation of ultramicroforms - extracellular vesicles from supernatants of Acholeplasma laidlawii PG8 (ubiquitous mycoplasma; the main contaminant of cell culture). Considering sizes, morphology, and ultrastructural organization, the ultramicroforms of A. laidlawii PG8 are similar to membrane vesicles of Gram positive and Gram-negative bacteria. We demonstrate that A. laidlawii PG8 vesicles contain genetic material and proteins, and are mutagenic to lymphocytes of human peripheral blood. We show that Mycoplasma gallisepticum S6, the other mycoplasma, also produce similar structures, which suggests that shedding of the vesicles might be the common phenomenon in Mollicutes. We found that the action of stress conditions results in the intensive formation of ultramicroforms in mycoplasmas. The role of vesicular formation in mycoplasmas remains to be studied. PMID- 21623459 TI - Defining the thrombotic risk in patients with myeloproliferative neoplasms. AB - Polycythemia vera (PV) and essential thrombocythemia (ET) are two Philadelphia negative myeloproliferative neoplasms (MPN) associated with an acquired mutation in the JAK2 tyrosine kinase gene. There is a rare incidence of progression to myelofibrosis and myeloid metaplasia in both disorders, which may or may not precede transformation to acute myeloid leukemia, but thrombosis is the main cause of morbidity and mortality. The pathophysiology of thrombosis in patients with MPN is complex. Traditionally, abnormalities of platelet number and function have been claimed as the main players, but increased dynamic interactions between platelets, leukocytes, and the endothelium do probably represent a fundamental interplay in generating a thrombophilic state. In addition, endothelial dysfunction, a well-known risk factor for vascular disease, may play a role in the thrombotic risk of patients with PV and ET. The identification of plasma markers translating the hemostatic imbalance in patients with PV and ET would be extremely helpful in order to define the subgroup of patients with a significant clinical risk of thrombosis. PMID- 21623460 TI - Asthma in sickle cell disease. AB - In recent years, evidence has increased that asthma predisposes to complications of sickle cell disease (SCD), such as pain crises, acute chest syndrome, pulmonary hypertension, and stroke, and is associated with increased mortality. An obstructive pattern of pulmonary function, along with a higher-than-expected prevalence of airway hyper-responsiveness (AHR) when compared to the general population, has led some researchers to suspect that underlying hemolysis may contribute to the development of a pulmonary disease similar to asthma in patients with SCD. While the pathophysiologic mechanism in atopic asthma involves up-regulation of Th2 cytokines, mast cell- and eosinophil-driven inflammation, plus increased activity of inducible nitric oxide synthase (iNOS) and arginase in airway epithelium resulting in obstructive changes and AHR, the exact mechanisms of AHR, obstructive and restrictive lung disease in SCD is unclear. It is known that SCD is associated with a proinflammatory state and an enhanced inflammatory response is seen during vaso-occlusive events (VOE). Hemolysis-driven acute-on chronic inflammation and dysregulated arginine-nitric oxide metabolism are potential mechanisms by which pulmonary dysfunction could occur in patients with SCD. In patients with a genetic predisposition of atopic asthma, these changes are probably more severe and result in increased susceptibility to sickle cell complications. Early recognition and aggressive management of asthma based on established National Institutes of Health asthma guidelines is recommended in order to minimize morbidity and mortality. PMID- 21623461 TI - The origins, specificity, and potential biological relevance of human anti-IgG hinge autoantibodies. AB - Human anti-IgG hinge (HAH) autoantibodies constitute a class of immunoglobulins that recognize cryptic epitopes in the hinge region of antibodies exposed after proteolytic cleavage, but do not bind to the intact IgG counterpart. Detailed molecular characterizations of HAH autoantibodies suggest that they are, in some cases, distinct from natural autoantibodies that arise independent of antigenic challenge. Multiple studies have attempted to define the specificity of HAH autoantibodies, which were originally detected as binding to fragments possessing C-terminal amino acid residues exposed in either the upper or lower hinge regions of IgGs. Numerous investigators have provided information on the isotype profiles of the HAH autoantibodies, as well as correlations among protease cleavage patterns and HAH autoantibody reactivity. Several biological functions have been attributed to HAH autoantibodies, ranging from house-cleaning functions to an immunosuppressive role to restoring function to cleaved IgGs. In this review, we discuss both the historic and current literature regarding HAH autoantibodies in terms of their origins, specificity, and proposed biological relevance. PMID- 21623462 TI - Effect of age on the association between body fat percentage and maximal walking speed. AB - OBJECTIVE: To study the effect of age on the association between body fat percentage and maximal walking speed in older people. DESIGN AND PARTICIPANTS: Cross-sectional analysis of data collected in the Finnish population-based Health 2000 Survey involving 916 men and 1 222 women aged 55 years and older with complete data on body composition and a walking speed test. METHODS: Body fat percentage was assessed using bioelectrical impedance analysis and maximal walking speed based on a timed walking test over a distance of 6.1 meters. Linear regression models were used to study the effect of age on association between body fat percentage and maximal walking speed. RESULTS: The association between body fat percentage quartiles and maximal walking speed differed significantly between persons of different ages (p for age interaction = 0.027). In the age stratified analyses, the association between body fat percentage and maximal walking speed remained significant among 60-69-year olds and 70-79-year-olds, but disappeared among 55-59-year-olds and 80-year and older after adjustment for potential covariates. Body fat percentage explained 11% of the variation in maximal walking speed among 55-59-year-olds, 21% among 60-69-year-olds, 17% among 70-79-year-olds and 11% among 80-year and older. CONCLUSION: Association between body fat percentage and maximal walking speed was strongest between the ages of 60 and 79 years. The results suggest that the effects of excess body fatness are especially harmful for physical functioning among adults in their sixties and seventies and they could benefit from interventions. PMID- 21623463 TI - Thigh muscle volume predicted by anthropometric measurements and correlated with physical function in the older adults. AB - OBJECTIVES: (1) to correlate thigh muscle volume measured by magnetic resonance image (MRI) with anthropometric measurements and physical function in elderly subjects; (2) to predict MRI-measured thigh muscle volume using anthropometric measurements and physical functional status in elderly subjects. DESIGN: Cross sectional, nonrandomized study. SETTING: Outpatient clinic in Taiwan. PARTICIPANTS: Sixty-nine elderly subjects (33 men and 36 women) aged 65 and older. MEASURMENTS: The anthropometric data (including body height, body weight, waist size, and thigh circumference), physical activity and function (including grip strength, bilateral quadriceps muscle power, the up and go test, chair rise, and five meters walk time) and bioelectrical impedance analysis data (including total body fat mass, fat-free mass, and predictive muscle size) were measured. MRI-measured muscle volume of both thighs was used as the reference standard. RESULTS: The MRI-measured thigh volume was positively correlated with all anthropometric data, quadriceps muscle power and the up and go test as well as fat-free mass and predictive muscle mass, whereas it was negatively associated with age and walk time. In predicting thigh muscle volume, the variables of age, gender, body weight, and thigh circumference were significant predictors in the linear regression model: Muscle volume (cm3) =4226.3-42.5 * Age (year)-955.7 * gender (male=1, female=2) + 45.9 * body weight(kg) + 60.0 * thigh circumference (cm) (r2 = 0.745, P < 0.001; standard error of the estimate = 581.6 cm3). CONCLUSION: The current work provides evidence of a strong relationship between thigh muscle volume and physical function in the elderly. We also developed a prediction equation model using anthropometric measurements. This model is a simple and noninvasive method for everyday clinical practice and follow-up. PMID- 21623464 TI - Body composition assessment of undernourished older subjects by dual-energy x-ray absorptiometry and bioelectric impedance analysis. AB - BACKGROUND: The prevention and treatment of diseases related to changes in body composition require accurate methods for the measurement of body composition. However, few studies have dealt specifically with the assessment of body composition of undernourished older subjects by different methodologies. OBJECTIVES: To assess the body composition of undernourished older subjects by two different methods, dual energy x-ray absorptiometry (DXA) and bioelectric impedance (BIA), and to compare results with those of an eutrophic group. DESIGN: The study model was cross-sectional; the study was performed at the University Hospital of the School of Medicine of Ribeirao Preto, University of Sao Paulo, Brazil. PARTICIPANTS: Forty-one male volunteers aged 62 to 91 years. The groups were selected on the basis of anamnesis, physical examination and nutritional assessment according to the Mini Nutritional Assessment (MNA) score. Body composition was assessed by DXA and BIA. RESULTS: Body weight, arm and calf circumference, body mass index (BMI), fat free mass (FFM) and fat mass (FM) were significantly lower in the undernourished group as compared to the eutrophic group. There were no significant differences between FFM and FM mean values determined by DXA and BIA in both groups, but the agreement between methods in the undernourished group was less strong. CONCLUSION: Our results suggest caution when BIA is to be applied in studies including undernourished older subjects. This study does not support BIA as an accurate method for the individual assessment of body composition. PMID- 21623465 TI - Improved diabetes control in the elderly delays global cognitive decline. AB - OBJECTIVES: To examine whether improved diabetes control is related to better cognitive outcomes. DESIGN: Randomized control trial. SETTING: A randomized trial of telemedicine vs. usual care in elderly persons with type 2 diabetes. PARTICIPANTS: Participants were 2169 persons 55 years and older with type 2 diabetes from New York City and Upstate New York. INTERVENTION: The diabetes case management intervention was implemented by a diabetes nurse, via a telemedicine unit in the participant's home, and in coordination with the primary care physician. MEASUREMENTS: Hemoglobin A1c (HbA1c), systolic blood pressure (SBP), and low density lipoprotein cholesterol (LDL), were measured at a baseline visit and at up to 5 annual follow-up visits. Global cognition was measured at those visits with the Comprehensive Assessment and Referral Evaluation (CARE). RESULT: In mixed models the intervention was related to slower global cognitive decline in the intervention group (p = 0.01). Improvements in HbA1c (p = 0.03), but not SBP or LDL, mediated the effect of the intervention on cognitive decline. CONCLUSION: Improved diabetes control in the elderly following existing guidelines through a telemedicine intervention was associated with less global cognitive decline. The main mediator of this effect seemed to be improvements in HbA1c. PMID- 21623466 TI - Sarcopenia: designing phase IIB trials. AB - Sarcopenia is the age-related involuntary loss of skeletal muscle mass and functionality that can lead to the development of disability, frailty and increased health care costs. The development of interventions aimed at preventing and/or treating sarcopenia is complex, requiring the adoption of assumptions and standards that are not well established scientifically or clinically. A number of investigators and clinicians (both from academia and industry) met in Rome (Italy) in 2009 to develop a consensus definition of sarcopenia. Subsequently, in Albuquerque (New Mexico, USA) in 2010, the same group met again to consider the complex issues necessary for designing Phase II clinical trials for sarcopenia. Current clinical trial data indicate that fat-free mass (FFM) parameters are responsive to physical activity/nutritional treatment modalities over short time periods, but pharmacological trials of sarcopenia have yet to show significant efficacy. In order to conduct a clinical trial within a reasonable time frame, groups that model or display accelerated aging and loss of FFM are necessary. Few studies have used acceptable designs for testing treatment effects, sample sizes or primary outcomes that could provide interpretable findings or effects across studies. Dual energy x-ray absorptiometry (DXA) is the measure of choice for assessing FFM, but sufficient time is needed for changes to be detected accurately and reliably. A tool set that would allow clinical, basic and epidemiological research on sarcopenia to advance rapidly toward diagnosis and treatment phases should be those reflecting function and strength. PMID- 21623467 TI - The effect of caloric restriction and glycemic load on measures of oxidative stress and antioxidants in humans: results from the CALERIE Trial of Human Caloric Restriction. AB - Decreasing oxidative stress and increasing antioxidant defense has been hypothesized as one mechanism by which caloric restriction (CR) increases longevity in animals. A total of 46 moderately overweight volunteers (BMI: 25-30 kg/m2), ages 20-42 yr were randomized to either high glycemic (HG) or low glycemic (LG) dietary load CR regimen at either 10% (n=12) or 30% (n=34) of basal caloric intake. All food was provided to participants for 6 mo. Overall, after controlling for CR levels and dietary regimen for 6 mo, plasma glutathione peroxidase activity increased (p=0.04) and plasma protein carbonyl levels decreased (p=0.02) and a non-significant decrease in plasma 8-epi-prostaglandin F2alpha level was observed (p=0.09). No significant change was observed in other plasma antioxidants such as superoxide dismutase and catalase. These findings indicate that short term CR (10% or 30%) in moderately overweight subjects modulates some but not all measures of antioxidant defense and oxidative stress. PMID- 21623468 TI - Similar effects of leucine rich and regular dairy products on muscle mass and functions of older polymyalgia rheumatica patients: a randomized crossover trial. AB - OBJECTIVES: Leucine-rich milk and whey proteins have been suggested for prevention of age related loss of muscle mass and strength i.e. sarcopenia. The effects of milk protein supplementation and low intensity home based physical exercise on body composition and muscle functions were investigated. DESIGN: A randomized double blind crossover trial. SETTING: Community dwelling members of Helsinki rheumatoid association. PARTICIPANTS: Older people (N=47, mean age 69.5 years) suffering from polymyalgia rheumatica. INTERVENTION: Patients performed as many stand ups as possible twice a day after which they ingested a regular (Control) or a whey protein enriched dairy product with high leucine content (Test). The 8-week intervention periods were separated by a 4-week wash-out. MEASUREMENTS: Body composition was measured by dual x-ray absorptiometry and muscle functions by hand grip strength, force platform countermovement jump performance, chair stand test, and walking speed. RESULTS: The 16-week home-based post-exercise supplementation resulted in a 1.8% increase (p = 0.052) in lower limb muscle mass. Walking speed (+5.3%, p = 0.007) and chair stand test performance (-12.2 %, p < 0.001) were also improved. Furthermore, a tendency for increased jump power (+3.0%, p = 0.084) was observed. However, significant and consistent differences were not found in the changes of muscle mass indices or muscle functions between supplements, but the test supplement tended to prevent accumulation of body fat. CONCLUSION: A low intensity home based exercise program combined with post-exercise milk protein supplementation is feasible despite some gastrointestinal complaints and seems effective in improving the muscle mass and functions of older persons with a inflammatory disease. Further studies are needed to establish, whether and to what extent the use of leucine-enriched whey products prevent or treat age-associated sarcopenia and whether they are superior to the present commercial milk products. PMID- 21623469 TI - Generalised and abdominal adiposity are important risk factors for chronic disease in older people: results from a nationally representative survey. AB - OBJECTIVES: To look at the trends in prevalence of generalised (body mass index (BMI) >= 25 kg/m2) and abdominal obesity (waist circumference (WC) >102 cm, men; > 88 cm, women) among older people from 1993 to 2008, prevalence of chronic disease by overweight/obesity and WC categories in England 2005 and evaluate the association of these measures with chronic diseases. DESIGN: Analyses of nationally representative cross-sectional population surveys, the Health Survey for England (HSE). PARTICIPANTS: Non-institutionalised men and women aged >= 65 years (in HSE 2005, 1512 men and 1747 women). MEASUREMENTS: Height, weight, waist circumference, blood pressure measurements were taken according to standardised HSE protocols. Information collected on socio-demographic, health behaviour and doctor diagnosed health conditions. RESULTS: Generalised obesity and abdominal obesity increased among men and women from 1993 to 2008. In 2005, the HSE 2005 focussed on older people. 72% of men and 68% of women aged over 65 were either overweight or obese. Prevalence of raised WC was higher in women (58%) than in men (46%). The prevalence of diabetes and arthritis was higher in people with generalised obesity in both sexes. Men were more likely to have had a joint replacement and had a higher prevalence of stroke if they were overweight only but women were more likely to have had a joint replacement only if they were obese (13%) and had a higher risk of falls with generalised obesity. The pattern was similar for the prevalence of chronic diseases by raised WC. Multivariate analysis showed that generalised and abdominal obesity was independently associated with risk of hypertension, diabetes and arthritis in both men and women. In women only, there was an association between generalised obesity and having a fall in the last year (OR: 1.5), and between abdominal obesity and having a joint replacement (OR: 1.9, p=0.01). CONCLUSION: Complications of obesity such as diabetes, hypertension and arthritis, are more common in men and women aged over 65 who are overweight or obese, as well as in those with a raised WC. These conditions impact on morbidity, mortality and have cost implications for the health service and are known to improve with weight loss even in old age. Treatment strategies to address these conditions such as weight management and prevention of overweight and obesity are important even in older people. There is a need to ensure that older people are given appropriate advice about keeping physically active and eating sensibly. PMID- 21623470 TI - The value of a formula including haematocrit, blood urea and gender (HUGE) as a screening test for chronic renal insufficiency. AB - INTRODUCTION: Despite increasing use in clinical practice, an estimated glomerular filtration rate value (eGFR) of <60 ml/min/1.73 m2 does not necessarily indicate the existence of chronic renal insufficiency (CRI) and this may lead to an over-estimate of CRI particularly in persons seventy years or older. AIM: To find a screening test able to differentiate CRI from the decrease in GFR normally associated with the renal ageing process. METHODS: Medical information of 487 individuals of both sexes aged 16-102 was obtained from nephrologists, internal medicine physicians, cardiologists, geriatricians, family and nuclear medicine doctors from Argentina, Portugal and Spain. Data were assessed and statistically analysed using logistic regression techniques. From the discriminative variables it was derived the HUGE formula. RESULTS: A formula including haematocrit , blood urea, and gender (HUGE), diagnoses CRI regardless of the variables of age, blood creatinine, creatinine clearance, or other eGFR. The HUGE formula is: L = 2.505458 - (0.264418 x Hematocrit) + (0.118100 x Urea) [+ 1.383960 if male]. If L is a negative number the individual does not have CRI; if L is a positive number, CRI is present. Our data demonstrate that the HUGE formula is more reliable than MDRD and CKD-EPI, particularly in persons aged over 70. CONCLUSIONS: Our HUGE screening formula offers a straightforward, easily available and inexpensive method for differentiating between CRI and eGFR < 60 ml/min/1.73 m2 that will prevent a considerable number of aged healthy persons, as much as 1.700.000 in Spain and 2.600.000 in U.K., to be excluded from clinical assays or treatments contraindicated in CRI. PMID- 21623471 TI - B vitamin status, dietary intake and length of stay in a sample of elderly rehabilitation patients. AB - OBJECTIVES: To investigate the relationships between previous diet, biomarkers of selected B vitamins, nutritional status and length of stay. DESIGN: Cross sectional study. SETTING: Geriatric rehabilitation patients, Sydney, Australia. PARTICIPANTS: Fifty two consenting patients with normal serum creatinine levels and no dementia. MEASUREMENTS: Serum vitamin B12, plasma vitamin B6, serum and erythrocyte folate, homocysteine and methylmalonic acid (MMA) concentrations; dietary intake using a validated semi-quantitative food frequency questionnaire and nutritional assessment using the Mini Nutritional Assessment (MNA). Length of stay data were collected from medical records after discharge. RESULTS: The age was 80 +/- 8 year (mean +/- SD), BMI 26.4 +/- 6.8 kg/m2 and MNA score 22 +/- 3 indicating some risk of malnutrition. Deficiencies of vitamins B6, B12 and folate were found in 30, 22 and 5 subjects respectively. Length of stay was positively correlated with age and MMA (Spearman's correlation 0.4, p<0.01 and 0.28, p<0.05 respectively) and negatively correlated with albumin, vitamin B6 and MNA score (Spearman's correlation -0.35, -0.33 and -0.29, p<0.05). After adjustment for age and sex, ln vitamin B6 and ln MMA concentrations were significant in predicting ln LOS (p=0.006 and p=0.014 respectively). CONCLUSION: The study indicates a high risk of vitamin B deficiencies in the elderly and suggests that deficiencies of vitamins B6 and B12 are associated with length of stay. This is concerning as B vitamin status is rarely fully assessed. PMID- 21623472 TI - Dizziness as a geriatric condition among rural community-dwelling older adults. AB - BACKGROUND: Dizziness is a common symptom in older adults. The majority of those with dizziness tend to have more than one risk factor, suggesting that dizziness is a multifactorial geriatric condition. Therefore, associated factors must be determined to permit risk-reduction approaches. OBJECTIVE: To examine the associations between dizziness and socio-demographic, physical, functional and psychological health factors among older persons living in the Andes Mountains. DESIGN: Population-based cross-sectional study. SETTINGS/PARTICIPANTS: One thousand six hundred ninety-two community-living people aged 60 years and over living in four rural and suburban areas of villages in coffee-grower zones in the Colombian Andes Mountains. MEASUREMENTS: Outcome measures included self-reporting of dizziness as a symptom experienced either very frequently or continuously during the last month. Independent variables were demographic, socioeconomic and social factors; disease and biomedical factors; functional status and performance based measures; and psychological factors such as depressive symptoms and self rated health. RESULTS: Dizziness was reported by 15.2% of participants in the study. Variables independently associated with dizziness were: number of chronic conditions, visual impairment, and use of more than four medications. Independently associated psychological variables were: poor self-perceived health, cognitive impairment and depression. Health and psychological factors accounted for 85% of dizziness. CONCLUSION: Older persons who reported dizziness were more physically frail, with more instances of chronic conditions and sensory impairments, and had poor self-perceptions of their health. Biomedical and psychological factors showed a strong independent association with dizziness. A multifactorial intervention targeting the identified factors would reduce dizziness in older people. However, this approach may need to address different sets of specific factors related to the dizziness categories. PMID- 21623474 TI - Smoking ban in the Alps--any wonder? PMID- 21623473 TI - Management of hepatitis C virus (HCV) infection in drug substitution programs. AB - BACKGROUND: In Switzerland, intravenous drug use (IDU) accounts for 80% of newly acquired hepatitis C virus (HCV) infections. Early HCV treatment has the potential to interrupt the transmission chain and reduce morbidity/mortality due to decompensated liver cirrhosis and hepatocellular carcinoma. Nevertheless, patients in drug substitution programs are often insufficiently screened and treated. OBJECTIVE/METHODS: With the aim to improve HCV management in IDUs, we conducted a cross sectional chart review in three opioid substitution programs in St. Gallen (125 methadone and 71 heroin recipients). Results were compared with another heroin substitution program in Bern (202 patients) and SCCS/SHCS data. RESULTS: Among the methadone/heroin recipients in St. Gallen, diagnostic workup of HCV was better than expected: HCV/HIV-status was unknown in only 1% (2/196), HCV RNA was not performed in 9% (13/146) of anti-HCV-positives and the genotype missing in 15% (12/78) of HCV RNA-positives. In those without spontaneous clearance (two thirds), HCV treatment uptake was 23% (21/91) (HIV-: 29% (20/68), HIV+: 4% (1/23)), which was lower than in methadone/heroin recipients and particularly non-IDUs within the SCCS/SHCS, but higher than in the, mainly psychiatrically focussed, heroin substitution program in Bern (8%). Sustained virological response (SVR) rates were comparable in all settings (overall: 50%, genotype 1: 35-40%, genotype 3: two thirds). In St. Gallen, the median delay from the estimated date of infection (IDU start) to first diagnosis was 10 years and to treatment was another 7.5 years. CONCLUSIONS: Future efforts need to focus on earlier HCV diagnosis and improvement of treatment uptake among patients in drug substitution programs, particularly if patients are HIV-co-infected. New potent drugs might facilitate the decision to initiate treatment. PMID- 21623475 TI - High prevalence of hypovitaminosis D in a Swiss rheumatology outpatient population. AB - Vitamin D is important for bone metabolism and neuromuscular function. While a routine dosage is often proposed in osteoporotic patients, it is not so evident in rheumatology outpatients where it has been shown that the prevalence of hypovitaminosis D is high. The aim of the current study was to systematically evaluate the vitamin D status in our outpatient rheumatology population to define the severity of the problem according to rheumatologic diseases. During November 2009, all patients were offered a screening test for 25-OH vitamin D levels and categorised as deficient (<10 ug/l [ng/ml] [25 nmol/l]), insufficient (10 ug/l to 30 ug/l [25 to 75 nmol/l]) or normal (>30 ug/l [75 nmol/l]). A total of 272 patients were included. The mean 25-OH vitamin D level was 21 ug/l (range 1.5 to 45.9). A total of 20 patients had vitamin D deficiency, 215 patients had an insufficiency and 37 patients had normal results. In the group of patients with osteoporosis mean level of 25-OH vitamin D was 25 ug/l and 31% had normal results. In patients with inflammatory rheumatic diseases (N = 219), the mean level of 25-OH vitamin D was 20.5 ug/l, and only 12% had normal 25-OH vitamin D levels. In the small group of patients with degenerative disease (N = 33), the mean level of 25-OH vitamin D was 21.8 ug/l, and 21% had normal results. Insufficiency and deficiency were even seen in 38% of the patients who were taking supplements. These results confirm that hypovitaminosis D is highly prevalent in an outpatient population of rheumatology patients, affecting 86% of subjects. Despite oral supplementation (taken in 38% of our population), only a quarter of those on oral supplementation attained normal values of 25-OH vitamin D. PMID- 21623476 TI - Do Swiss adolescents perceive the negative effects of their illegal substance use? AB - This study explores adolescents' perceptions of adverse consequences linked to their illegal psychoactive substance (IPS) use, as they are often thought to minimise them. From a Swiss nationally representative sample of 8740 adolescents aged 16 to 20 pursuing post-mandatory education, 2515 participants reported IPS use in the past month on a self-administered anonymous questionnaire. The percentages of participants reporting problems in four areas (individual, school, relationships and sexual) were assessed, depending on the type of IPS consumption over the last 30 days: occasional cannabis users: <=2 times; regular cannabis users: >=3 times; and poly-consumers: cannabis plus at least one other substance used. The percentages varied significantly across these three groups with 26.9% of occasional users, 53.8% of regular users, and 73.3% of poly-consumers reporting at least one problem. Compared to occasional cannabis users, poly consumers were more likely to report problems in all four categories [relative risk ratio (RRR): 3.38 to 5.44], while regular cannabis users often reported only school and relationship problems [RRR: 2.43 to 3.23]. Thus, many adolescents seem to perceive the negative effects of their IPS use, with heavier consumption being associated with increasing problems. Physicians should feel confident questioning adolescents on the adverse consequences of their IPS consumption, as they are likely to be responsive on this issue. PMID- 21623477 TI - Incidence of acute myocardial infarction after implementation of a public smoking ban in Graubunden, Switzerland: two year follow-up. AB - QUESTION UNDER STUDY: In the first year after implementation of a public smoking ban a significant decrease in the incidence of acute myocardial infarction (AMI) was observed in Graubunden. In the present study we analyzed the incidence of AMI in the second year of the ban. In addition, we investigated the contribution of smoking ban-unrelated factors to the reduced incidence of AMI incidence observed after enactment of the ban. METHODS: Data of all AMI patients who underwent coronary angiography at the Kantonsspital Graubunden, the only tertiary care hospital with a cardiac catheterization laboratory in Graubunden, between March 1st, 2009 and February 28th, 2010 were collected prospectively. Data were compared with those of the three preceding 12-month periods. We also estimated AMI incidence during the corresponding time period in Lucerne, a region with no smoke-free legislation, using data of the AMIS Plus registry. The influence of outdoor air pollution was analyzed with the help of official measurements of PM(10)- and NO(2)-concentrations in Graubunden. The prescription of lipid lowering drugs was estimated by using sales figures in Graubunden and Lucerne. RESULTS: In Graubunden, the number of patients with AMI in the second year after adoption of the smoking ban was similar to that in the first year of the ban (188 vs. 183; P = ns) and significantly lower than in each of the two years preceding the ban (229 and 242, respectively; P <0.05 vs. each of the 12-month periods after the ban). Overall, the number of AMI patients in the two post-ban years was 21% lower than in the two pre-ban years. The reduction in the number of patients with AMI was most pronounced in non-smokers and individuals with known coronary artery disease. During the corresponding time period, no similar decrease in the incidence of AMI was observed in Lucerne. No association was found between the magnitude of outdoor air pollution and the incidence of AMI. During the observation period, the use of lipid-lowering drugs increased similarly in Graubunden and Lucerne. CONCLUSIONS: Compared with the two years preceding the implementation of a smoking ban, the incidence of AMI remained significantly reduced in the second year of the ban in Graubunden, whereas no similar reduction was seen in a comparable area without smoke-free legislation. Changes in outdoor air pollution or the use of lipid-lowering drugs did not substantially contribute to the decrease in the incidence of AMI that occurred after adoption of the ban in Graubunden. PMID- 21623478 TI - The time-course of threat processing in children: a temporal dissociation between selective attention and behavioral interference. AB - Although selective attention to threatening information is an adaptive mechanism, exaggerated attention to threat may be related to anxiety disorders. However, studies examining threat processing in children have obtained mixed findings. In the present study, the time-course of attentional bias for threat and behavioral interference was analyzed in a community sample of 8-18-year-old children (N=33) using a pictorial dot probe task. Threatening and neutral stimuli were shown during 17 ms (masked), 500 ms, and 1250 ms. Results provide preliminary evidence of an automatic attentional bias for threat at 17 ms that persists during later, more controlled stages of information processing (500 and 1250 ms). Furthermore, participants showed a delayed response to threat-containing trials relative to neutral trials in the 500 and 1250 ms condition, which may indicate interference by threat. Together, these results suggest that an attentional bias for threat precedes behavioral interference in children. Furthermore, results indicate that performance in daily life can be temporarily interrupted by the processing of threatening information. In addition, results of earlier studies into selective attention in children using tasks based on behavioral responses may have been confounded by interference effects of threat. For future studies, we recommend to take behavioral interference into account. PMID- 21623479 TI - Social factors and mental health symptoms among women who have experienced involuntary job loss. AB - The growing proportion of women in the labor force and the current economic crisis has made women a target population for job loss. In that situation, they are likely to experience recurrent layoffs, which force them to cope with multiple job loss and with unemployment. The present study aimed to examine sociodemographic factors that help women succeed in coping with single or multiple experiences of job loss, and that even enhance their self-efficacy in returning to work. The population of participants consisted of 134 Israeli women aged 30-45 who had been laid off. The findings indicate that the women who had experienced multiple job loss expressed a stronger desire to return to work than did those who had been laid off only once. Married women showed a greater tendency to become accustomed to stress after being laid off than did never married women. However, even though multiple job loss might be a forced solution to home-work conflict, never-married women were found to be at risk for distress responses after being laid off. There is a need to develop responses for women who are at risk for multiple job loss, and to enhance employers' awareness of the situation faced by women who are laid off. PMID- 21623480 TI - The dual-effects model of social control revisited: relationship satisfaction as a moderator. AB - The dual-effects model of social control states that receiving social control leads to better health behavior, but also enhances distress in the control recipient. Associated findings, however, are inconsistent. In this study we investigated the role of relationship satisfaction as a moderator of associations of received spousal control with health behavior and affect. In a study with five waves of assessment spanning two weeks to one year following radical prostatectomy (RP), N=109 married or cohabiting prostate-cancer patients repeatedly reported on their pelvic-floor exercise (PFE) to control postsurgery urinary incontinence and affect as primary outcomes, on received PFE-specific spousal control, relationship satisfaction, and covariates. Findings from two level hierarchical linear models with repeated assessments nested in individuals suggested significant interactions of received spousal control with relationship satisfaction predicting patients' concurrent PFE and positive affect. Patients who were happy with their relationships seemed to benefit from spousal control regarding regular PFE postsurgery while patients less satisfied with their relationships did not. In addition, the latter reported lower levels of positive affect when receiving much spousal control. Results indicate the utility of the inclusion of relationship satisfaction as a moderator of the dual-effects model of social control. PMID- 21623481 TI - Competitive protein adsorption on polysaccharide and hyaluronate modified surfaces. AB - Adsorption of bovine serum albumin (BSA) and fibrinogen (Fg) was measured on six distinct bare and dextran- and hyaluronate-modified silicon surfaces created using two dextran grafting densities and three hyaluronic acid (HA) sodium salts derived from human umbilical cord, rooster comb and Streptococcus zooepidemicus. Film thickness and surface morphology depended on the HA molecular weight and concentration. BSA coverage was enhanced on surfaces in competitive adsorption of BSA:Fg mixtures. Dextranization differentially reduced protein adsorption onto surfaces based on oxidation state. Hyaluronization was demonstrated to provide the greatest resistance to protein coverage, equivalent to that of the most resistant dextranized surface. Resistance to protein adsorption was independent of the type of HA utilized. With changing bulk protein concentration from 20 to 40 MUg ml(-1) for each species, Fg coverage on silicon increased by 4x, whereas both BSA and Fg adsorption on dextran and HA were far less dependent on protein bulk concentration. PMID- 21623483 TI - Working memory training improves reading processes in typically developing children. AB - The goal of this study was to investigate whether a brief cognitive training intervention results in a specific performance increase in the trained task, and whether there are transfer effects to other nontrained measures. A computerized, adaptive working memory intervention was conducted with 9- to 11-year-old typically developing children. The children considerably improved their performance in the trained working memory task. Additionally, compared to a matched control group, the experimental group significantly enhanced their reading performance after training, providing further evidence for shared processes between working memory and reading. PMID- 21623482 TI - Atomic force microscopy investigations of heterogeneities in the adhesion energies measured between pathogenic and non-pathogenic Listeria species and silicon nitride as they correlate to virulence and adherence. AB - Atomic force microscopy (AFM) was used to probe heterogeneities in adhesion energies measured between pathogenic and non-pathogenic species of Listeria and silicon nitride in water at four levels. Adhesion energies were quantified on individual bacterial cells (cell level), bacterial cells that belonged to an individual Listeria strain but varied in their cultures (strain level), bacterial cells that belonged to an individual Listeria species but varied in their strain type (species level) and on bacterial cells that belonged to the Listeria genus but varied in their species type (genus level). To quantify heterogeneities in the adhesion energies, a heterogeneity index (HI) was defined based on quantified standard errors of mean. At the cell level, spatial variations in the adhesion energies were not observed. For the strain, species, and genus levels, the HI increased with increased adhesion energies. At the species level, the HI increased with strain virulence. PMID- 21623484 TI - Counting to ten milliseconds: low-anger, but not high-anger, individuals pause following negative evaluations. AB - Low-anger individuals are less reactive, both emotionally and behaviourally, to a large variety of situational primes to anger and aggression. Why this is so, from an affective processing perspective, has been largely conjectural. Four studies (total N=270) sought to link individual differences in anger to tendencies exhibited in basic affective processing tasks. On the basis of motivational factors and considerations, it was hypothesised that negative evaluations would differentially activate a psychological alarm system at low levels of anger, resulting in a pause that should be evident in the speed of making subsequent evaluations. Just such a pattern was evident in all studies. By contrast, high anger individuals did not pause following their negative evaluations. In relation to this affective processing tendency, at least, dramatically different effects were observed among low- versus high-anger individuals. Implications for the personality-processing literature, theories of trait anger, and fast-acting regulatory processes are discussed. PMID- 21623485 TI - Does ignoring lead to worse evaluations? A new explanation of the stimulus devaluation effect. AB - Affective evaluation of stimuli just seen in visual search tasks has been shown to depend on task-relevant stimulus configuration (Raymond, Fenske, & Tavassoli, 2003): Whereas targets and novel stimuli were evaluated similarly, distractors were devaluated. These results were explained by an inhibition-based account of the influence of selective attention on emotion. In the present experiments, we demonstrated that stimulus devaluation might not be a consequence of attentional inhibition. By simply instructing participants to react to an accepted or rejected stimulus in the visual search task of Experiment 1, we found distractor devaluation in the first case and target devaluation in the second case. We conclude that devaluation of stimuli is mediated by the affective connotation implied by response labels and instructions. This was confirmed in Experiment 2: To-be-ignored stimuli were not devaluated when participants knew that those stimuli would become task-relevant during the experiment. PMID- 21623486 TI - Affective state and event-based prospective memory. AB - Event-based prospective memory tasks require the realisation of a delayed intention at the occurrence of a specific target event. The present research investigates how performance in this kind of prospective memory task is influenced by the current affective state. By manipulating participants' mood during intention realisation we tested two competing models of mood effects on memory (i.e., a capacity consuming account and a processing style account). Furthermore, we manipulated the valence of the target event to investigate mood congruency effects in prospective memory. No evidence was found for a mood congruency effect, but the results showed that prospective memory performance increased with a sad mood. This effect is consistent with recent theories on mood dependent processing-style regulation, postulating that a sad mood produces a more analytic and detailed processing style whereas a happy mood produces a more global and less detailed processing style. PMID- 21623487 TI - "That's not my arm": a hypnotic analogue of somatoparaphrenia. AB - INTRODUCTION: "Instrumental hypnosis" allows researchers to model clinical symptoms in the laboratory, creating "virtual patients" with reversible disturbances in, for example, perception, action, memory, or belief. We used hypnosis to temporarily recreate somatoparaphrenia, a delusional belief that one's own limb belongs to someone else. METHODS: We compared a "Fully Formed" somatoparaphrenia suggestion with a "Factor 1 + Factor 2" suggestion that attempted to generate the delusional belief from analogues of its hypothesised underlying factors (i.e., paralysis plus disrupted critical belief evaluation). We tested and then challenged subjects' responses to these suggestions. RESULTS: Although many hypnotic subjects experienced temporary paralysis, only a minority claimed their arm did not belong to them. Notably, the Fully Formed suggestion was more successful in recreating features of somatoparaphrenia than the Factor 1 + Factor 2 suggestion. In response to the challenges, some of those who developed temporary somatoparaphrenia maintained their belief throughout the hypnosis session. CONCLUSIONS. We discuss these findings in terms of the "two-factor" theory of delusions and we highlight the advantages versus disadvantages of using hypnosis to explore such delusional beliefs in the laboratory. PMID- 21623488 TI - Association between locus of control in childhood and psychotic symptoms in early adolescence: results from a large birth cohort. AB - INTRODUCTION. Specific attributional styles have been demonstrated in individuals with psychotic disorders and are implicated in the development of psychotic symptoms. We aimed to examine the association between locus of control (LOC) assessed in childhood and psychotic symptoms reported in early adolescence. METHODS. We used a prospective longitudinal design using data from a large birth cohort (the Avon Longitudinal Study of Parents and Children, ALPSAC). 6455 subjects completed a semistructured clinical interview assessing 12 individual psychotic symptoms at a mean age of 12.9 years. A measure of LOC was previously collected in the cohort at the age of 8. RESULTS. Children who reported an externalised LOC at age 8 were at increased risk of reporting both broadly defined (OR 1.77, 95% CI 1.49 to 2.08) and narrowly defined (OR 2.06, 95% CI 1.58 to 2.67) psychotic symptoms at age 13 years. These associations were only slightly attenuated after adjustment for potential confounders. The associations were similar for broadly defined specific paranoid symptoms but weaker for narrowly defined specific paranoid symptoms. CONCLUSIONS. An externalised LOC appears to be associated with later reporting of psychotic symptoms in early adolescence. Further investigation of the role of attributional styles, such as LOC, in increasing the risk for psychotic disorders, is warranted. PMID- 21623490 TI - Health Canada: current topics in food chemical safety research. PMID- 21623489 TI - Present and future directions of translational research on aflatoxin and hepatocellular carcinoma. A review. AB - The aflatoxins were discovered in toxic peanut meal causing "turkey X" disease, which killed large numbers of turkey poults, ducklings and chicks in the UK in the early 1960s. Extracts of toxic feed induced the symptoms in experimental animals, and purified metabolites with properties identical to aflatoxins B(1) and G(1) (AFB(1) and AFG(1)) were isolated from Aspergillus flavus cultures. Structure elucidation of aflatoxin B(1) was accomplished and confirmed by total synthesis in 1963. AFB(1) is a potent liver carcinogen in rodents, non-human primates, fish and birds, operating through a genotoxic mechanism involving metabolic activation to an epoxide, formation of DNA adducts and, in humans, modification of the p53 gene. Aflatoxins are unique among environmental carcinogens, in that elucidation of their mechanisms of action combined with molecular epidemiology provides a foundation for quantitative risk assessment; extensive evidence confirms that contamination of the food supply by AFB(1) puts an exposed population at increased risk of developing hepatocellular carcinoma (HCC). Molecular biomarkers to quantify aflatoxin exposure in individuals were essential to link aflatoxin exposure with liver cancer risk. Biomarkers were validated in populations with high HCC incidence in China and The Gambia, West Africa; urinary AFB(1)-N (7)-Guanine excretion was linearly related to aflatoxin intake, and levels of aflatoxin-serum albumin adducts also reflected aflatoxin intake. Two major cohort studies employing aflatoxin biomarkers identified their causative role in HCC etiology. Results of a study in Shanghai men strongly support a causal relationship between HCC risk and the presence of biomarkers for aflatoxin and HBV infection, and also show that the two risk factors act synergistically. Subsequent cohort studies in Taiwan confirm these results. IARC classified aflatoxin as a Group 1 human carcinogen in 1993, based on sufficient evidence in humans and experimental animals indicating the carcinogenicity of naturally occurring mixtures of aflatoxins, aflatoxin B(1), G(1) and M(1). Aflatoxin biomarkers have also been used to show that primary prevention to reduce aflatoxin exposure can be achieved by low-technology approaches at the subsistence farm level in sub-Saharan Africa. Also, in residents of Qidong, China, oral dosing with chlorophyllin, a chlorophyll derivative, prior to each meal led to significant reduction in aflatoxin-DNA biomarker excretion, supporting the feasibility of preventive measures to reduce HCC risk in populations experiencing unavoidable aflatoxin exposure. The systematic, comprehensive approach used to create the total aflatoxin database justifies optimism for potential success of preventive interventions to ameliorate cancer risk attributable to aflatoxin exposure. This strategy could serve as a template for the development, validation and application of molecular and biochemical markers for other carcinogens and cancers as well as other chronic diseases resulting from environmental exposures. PMID- 21623491 TI - Health Canada's Food Directorate. Introduction. PMID- 21623492 TI - Determination of melamine, ammeline, ammelide and cyanuric acid in infant formula purchased in Canada by liquid chromatography-tandem mass spectrometry. AB - A liquid chromatography-tandem mass spectrometry-based isotope dilution method was developed for the analysis of the triazine compounds melamine (MEL), ammeline (AMN), ammelide (AMD) and cyanuric acid (CYA) in infant formula samples purchased in Canada in 2008 for the purpose of a combined exposure and risk assessment. Infant formula samples were extracted with 1:1 acetonitrile-water, cleaned up on disposable ion-exchange solid-phase extraction cartridges, and analysed by ultra high-performance liquid chromatography-tandem mass spectrometry. MEL and CYA were detected in almost all infant formula products: the highest concentrations observed were 0.32 mg kg(-1) MEL and 0.45 mg kg(-1) CYA. Samples that were relatively high in MEL in this survey tended to be low in CYA, and vice versa. Concentrations of AMN and AMD were very low in all samples. The total of MEL related compounds (sum of all four analytes) in all samples was below the interim standard of 0.5 mg kg(-1) for infant formula products established by Health Canada. PMID- 21623493 TI - Gluten contamination in the Canadian commercial oat supply. AB - A growing body of evidence suggests that a majority of people with celiac disease and on a gluten-free diet can safely consume pure oats in moderate amounts; however, previous studies have indicated that the commercial oat supply in other countries, and in Canada to some extent, is contaminated with other grains. This study has confirmed that the commercial oat supply in Canada is heavily contaminated with gluten from other grains. Approximately 88% of the oat samples (n = 133) were contaminated above 20 mg kg(-1) and there were no differences between the oat types tested. Only one gluten-free variety of oats was analysed and it consistently provided negative results in all analyses. It is difficult to determine where the contamination originates, but there are possibilities for cross-contamination in the field, in the transport of the grain, in the storage of the grain, and in the milling and packaging facilities. It is clear from this study that only those products that have been certified 'pure' oats would be appropriate for a gluten-free diet. PMID- 21623494 TI - Paralytic shellfish poisoning (PSP) toxin binders for optical biosensor technology: problems and possibilities for the future: a review. AB - This review examines the developments in optical biosensor technology, which uses the phenomenon of surface plasmon resonance, for the detection of paralytic shellfish poisoning (PSP) toxins. Optical biosensor technology measures the competitive biomolecular interaction of a specific biological recognition element or binder with a target toxin immobilised onto a sensor chip surface against toxin in a sample. Different binders such as receptors and antibodies previously employed in functional and immunological assays have been assessed. Highlighted are the difficulties in detecting this range of low molecular weight toxins, with analogues differing at four chemical substitution sites, using a single binder. The complications that arise with the toxicity factors of each toxin relative to the parent compound, saxitoxin, for the measurement of total toxicity relative to the mouse bioassay are also considered. For antibodies, the cross-reactivity profile does not always correlate to toxic potency, but rather to the toxin structure to which it was produced. Restrictions and availability of the toxins makes alternative chemical strategies for the synthesis of protein conjugate derivatives for antibody production a difficult task. However, when two antibodies with different cross-reactivity profiles are employed, with a toxin chip surface generic to both antibodies, it was demonstrated that the cross reactivity profile of each could be combined into a single-assay format. Difficulties with receptors for optical biosensor analysis of low molecular weight compounds are discussed, as are the potential of alternative non-antibody based binders for future assay development in this area. PMID- 21623495 TI - Formation of acrylamide at temperatures lower than 100 degrees C: the case of prunes and a model study. AB - Acrylamide concentrations in prune products--baby strained prunes (range = 75-265 ug kg(-1)), baby apple/prune juice (33-61 ug kg(-1)), prune juice (186-916 ug kg( 1)) and prunes (58-332 ug kg(-1))--on the Canadian market were determined. The formation of acrylamide in a simulated plum juice was also investigated under 'drying conditions' in an open vessel at temperatures <100 degrees C for 24 h and under 'wet conditions' in a closed vessel at a temperature of 120 degrees C for 1 h. Acrylamide was produced in a simulated plum juice under 'drying conditions' in amounts comparable with those found in prunes and prune juices. Acrylamide was not produced in simulated plum juice under 'wet conditions' in a closed vessel at temperature of 120 degrees C for 1 h, but under the same condition an authentic prune juice doubled its acrylamide concentration. Formation of acrylamide in prune products was attributed to the presence of asparagine and sugars in the starting materials. PMID- 21623496 TI - Design and characterization of a direct ELISA for the detection and quantification of leucomalachite green. AB - Malachite green (MG), a member of the N-methylated triphenylmethane class of dyes, has long been used to control fungal and protozoan infections in fish. MG is easily absorbed by fish during waterborne exposure and is rapidly metabolized into leucomalachite green (LMG), which is known for its long residence time in edible fish tissue. This paper describes the development of an enzyme-linked immunosorbent assay (ELISA) for the detection and quantification of LMG in fish tissue. This development includes a simple and versatile method for the conversion of LMG to monodesmethyl-LMG, which is then conjugated to bovine serum albumin (BSA) to produce an immunogenic material. Rabbit polyclonal antibodies are generated against this immunogen, purified and used to develop a direct competitive enzyme-linked immunosorbent assay (ELISA) for the screening and quantification of LMG in fish tissue. The assay performed well, with a limit of detection (LOD) and limit of quantification (LOQ) of 0.1 and 0.3 ng g(-1) of fish tissue, respectively. The average extraction efficiency from a matrix of tilapia fillets was approximately 73% and the day-to-day reproducibility for these extractions in the assay was between 5 and 10%. PMID- 21623497 TI - Levels of total mercury in predatory fish sold in Canada in 2005. AB - Total mercury was analysed in 188 samples of predatory fish purchased at the retail level in Canada in 2005. The average concentrations (ng g(-1), range) were: sea bass 329 (38-1367), red snapper 148 (36-431), orange roughy 543 (279 974), fresh water trout 55 (20-430), grouper 360 (8-1060), black cod 284 (71 651), Arctic char 37 (28-54), king fish 440 (42-923), tilefish 601 (79-1164) and marlin 854 (125-2346). The Canadian standard for maximum total mercury allowed in the edible portions of fish sold at the retail level is 1000 ng g(-1) for shark, swordfish, marlin, orange roughy, escolar and both fresh and frozen tuna. The standard is 500 ng g(-1) for all other types of fish. In this study, despite the small number of samples of each species, the 1000 ng g(-1) maximum was exceeded in five samples of marlin (28%). The 500 ng g(-1) maximum was exceeded by six samples of sea bass (20%), four of tilefish (50%), five of grouper (24%), six of king fish (40%) and one of black cod (13%). PMID- 21623498 TI - Lead, cadmium and aluminum in Canadian infant formulae, oral electrolytes and glucose solutions. AB - Lead (Pb), cadmium (Cd) and aluminum (Al) were determined in 437 individual samples of infant formulae, oral electrolytes and 5% glucose solutions available in Canada. In the electrolytes, Cd and Pb concentrations were all below 0.01 and 0.041 ng g(-1), respectively. In the 5% glucose solutions, Pb and Cd levels averaged 0.01 and 0.09 ng g(-1), respectively. Reported on an as-consumed basis, Pb levels in milk- and soya-based formulae averaged 0.90 and 1.45 ng g(-1), respectively, while Cd levels averaged 0.23 and 1.18 ng g(-1), respectively Average Al levels on an as-consumed basis were 440 ng g(-1) (range 10-3400 ng g( 1)) in milk-based formulae and 730 ng g(-1) (range 230-1100 ng g(-1)) in soy based formulae. Al concentrations increased in the following order: plain formula < low-iron formula < iron-supplemented formula < casein hydrolysate formula ~ premature formula <= soy formula. For example, in the powdered formulae, average Al concentrations were 18 ng g(-1) for plain milk-based, 37 ng g(-1) for low iron, 128 ng g(-1) for iron supplemented, 462 ng g(-1) for lactose-free, 518 ng g(-1) for hypoallergenic and 619 ng g(-1) for soy-based formula. Al concentrations, as-consumed, increased with decreasing levels of concentration: powder < concentrated liquid < ready-to-use. Formulae stored in glass bottles contained between 100 and 300 ng g(-1) more Al than the same formulae stored in cans. The source of the increased Al did not appear to be the glass itself, because most electrolytes and glucose solutions, also stored in glass, contained less than 8 ng g(-1) Al. Corresponding differences in Pb and Cd levels were not observed. Al concentrations varied substantially among manufacturers; however, all manufacturers were able to produce plain milk-based formulae containing less than 50 ng g(-1) Al, i.e. within the range of Al concentrations found in human milk. Next to soya-based and hypoallergenic formulae, premature formulae contained among the highest concentrations of Al, ranging 851-909 ng g(-1) from one manufacturer and 365-461 ng g(-1) from another. PMID- 21623499 TI - Application of isotope dilution mass spectrometry: determination of ochratoxin A in the Canadian Total Diet Study. AB - Analytical methods are generally developed and optimized for specific commodities. Total Diet Studies, representing typical food products 'as consumed', pose an analytical challenge since every food product is different. In order to address this technical challenge, a selective and sensitive analytical method was developed suitable for the quantitation of ochratoxin A (OTA) in Canadian Total Diet Study composites. The method uses an acidified solvent extraction, an immunoaffinity column (IAC) for clean-up, liquid chromatography tandem mass spectrometry (LC-MS/MS) for identification and quantification, and a uniformly stable isotope-labelled OTA (U-[(13)C(20)]-OTA) as an internal recovery standard. Results are corrected for this standard. The method is accurate (101% average recovery) and precise (5.5% relative standard deviation (RSD)) based on 17 duplicate analysis of various food products over 2 years. A total of 140 diet composites were analysed for OTA as part of the Canadian Total Diet Study. Samples were collected at retail level from two Canadian cities, Quebec City and Calgary, in 2008 and 2009, respectively. The results indicate that 73% (102/140) of the samples had detectable levels of OTA, with some of the highest levels of OTA contamination found in the Canadian bread supply. PMID- 21623500 TI - Ochratoxin A in cocoa and chocolate sampled in Canada. AB - In order to determine the levels of ochratoxin A (OTA) in cocoa and cocoa products available in Canada, a previously published analytical method, with minor modifications to the extraction and immunoaffinity clean-up and inclusion of an evaporation step, was initially used (Method I). To improve the low method recoveries (46-61%), 40% methanol was then included in the aqueous sodium bicarbonate extraction solvent (pH 7.8) (Method II). Clean-up was on an OchratestTM immunoaffinity column and OTA was determined by liquid chromatography (LC) with fluorescence detection. Recoveries of OTA from spiked cocoa powder (0.5 and 5 ng g(-1)) were 75-84%; while recoveries from chocolate were 93-94%. The optimized method was sensitive (limit of quantification (LOQ) = 0.07-0.08 ng g( 1)), accurate (recovery = 75-94%) and precise (coefficient of variation (CV) < 5%). It is applicable to cocoa and chocolate. Analysis of 32 samples of cocoa powder (16 alkalized and 16 natural) for OTA showed an incidence of 100%, with concentrations ranging from 0.25 to 7.8 ng g(-1); in six samples the OTA level exceeded 2 ng g(-1), the previously considered European Union limit for cocoa. The frequency of detection of OTA in 28 chocolate samples (21 dark or baking chocolate and seven milk chocolate) was also 100% with concentrations ranging from 0.05 to 1.4 ng g(-1); one sample had a level higher than the previously considered European Union limit for chocolate (1 ng g(-1)). PMID- 21623502 TI - Sampling of cereals and cereal-based foods for the determination of ochratoxin A: an overview. AB - The mycotoxin ochratoxin A (OTA) is known to be heterogeneously distributed both intrinsically (from one individual food item to the next) as well as distributionally (throughout a sample of individual food items) in cereals and cereal-based foods. Therefore, proper sampling and sample comminution are special challenges, but are prerequisites for obtaining sound analytical data. This paper outlines the issue of the sampling process for cereals and cereal-based foods, starting with the planning phase, followed by the sampling step itself and the formation of analytical samples. The sampling of whole grain and retail-level cereal-based foods will be discussed. Furthermore, possibilities to reduce sampling variance are presented. PMID- 21623501 TI - Surveys of rice sold in Canada for aflatoxins, ochratoxin A and fumonisins. AB - Approximately 200 samples of rice (including white, brown, red, black, basmati and jasmine, as well as wild rice) from several different countries, including the United States, Canada, Pakistan, India and Thailand, were analysed for aflatoxins, ochratoxin A (OTA) and fumonisins by separate liquid chromatographic methods in two different years. The mean concentrations for aflatoxin B(1) (AFB(1)) were 0.19 and 0.17 ng g(-1) with respective positive incidences of 56% and 43% (>= the limit of detection (LOD) of 0.002 ng g(-1)). Twenty-three samples analysed in the second year also contained aflatoxin B(2) (AFB(2)) at levels >=LOD of 0.002 ng g(-1). The five most contaminated samples in each year contained 1.44-7.14 ng AFB(1) g(-1) (year 1) and 1.45-3.48 ng AFB(1) g(-1) (year 2); they were mostly basmati rice from India and Pakistan and black and red rice from Thailand. The average concentrations of ochratoxin A (OTA) were 0.05 and 0.005 ng g(-1) in year 1 and year 2, respectively; incidences of samples containing >=LOD of 0.05 ng g(-1) were 43% and 1%, respectively, in the 2 years. All positive OTA results were confirmed by LC-MS/MS. For fumonisins, concentrations of fumonisin B(1) (FB(1)) averaged 4.5 ng g(-1) in 15 positive samples (>=0.7 ng g(-1)) from year 1 (n = 99); fumonisin B(2) (FB(2)) and fumonisin B(3) (FB(3)) were also present (>=1 ng g(-1)). In the second year there was only one positive sample (14 ng g(-1) FB(1)) out of 100 analysed. All positive FB(1) results were confirmed by LC-MS/MS. PMID- 21623503 TI - Determination of the cyanobacterial toxin cylindrospermopsin in algal food supplements. AB - For the analysis of blue-green algal food supplements for cylindrospermopsin (CYN), a C18 solid-phase extraction column and a polygraphitized carbon solid phase extraction column in series was an effective procedure for the clean-up of extracts. Determination of CYN was by liquid chromatography with ultraviolet light detection. At extract spiking levels of CYN equivalent to 25-500 ug g(-1), blue-green algal supplement recoveries were in the range 70-90%. CYN was not detected in ten samples of food supplements and one chocolate product, all containing blue-green algae. The limit of detection for the method was 16 ug g( 1), and the limit of quantification was 52 ug g(-1). PMID- 21623504 TI - Concentrations of bisphenol A in the composite food samples from the 2008 Canadian total diet study in Quebec City and dietary intake estimates. AB - A total of 154 food composite samples from the 2008 total diet study in Quebec City were analysed for bisphenol A (BPA), and BPA was detected in less than half (36%, or 55 samples) of the samples tested. High concentrations of BPA were found mostly in the composite samples containing canned foods, with the highest BPA level being observed in canned fish (106 ng g(-1)), followed by canned corn (83.7 ng g(-1)), canned soups (22.2-44.4 ng g(-1)), canned baked beans (23.5 ng g(-1)), canned peas (16.8 ng g(-1)), canned evaporated milk (15.3 ng g(-1)), and canned luncheon meats (10.5 ng g(-1)). BPA levels in baby food composite samples were low, with 2.75 ng g(-1) in canned liquid infant formula, and 0.84-2.46 ng g(-1) in jarred baby foods. BPA was also detected in some foods that are not canned or in jars, such as yeast (8.52 ng g(-1)), baking powder (0.64 ng g(-1)), some cheeses (0.68-2.24 ng g(-1)), breads and some cereals (0.40-1.73 ng g(-1)), and fast foods (1.1-10.9 ng g(-1)). Dietary intakes of BPA were low for all age-sex groups, with 0.17-0.33 ug kg(-1) body weight day(-1) for infants, 0.082-0.23 ug kg(-1) body weight day(-1) for children aged from 1 to 19 years, and 0.052-0.081 ug kg(-1) body weight day(-1) for adults, well below the established regulatory limits. BPA intakes from 19 of the 55 samples account for more than 95% of the total dietary intakes, and most of the 19 samples were either canned or in jars. Intakes of BPA from non-canned foods are low. PMID- 21623505 TI - Determination of perchlorate in infant formula by isotope dilution ion chromatography/tandem mass spectrometry. AB - A sensitive and selective isotope dilution ion chromatography/tandem mass spectrometry (ID IC-MS/MS) method was developed and validated for the determination of perchlorate in infant formula. The perchlorate was extracted from infant formula by using 20 ml of methanol and 5 ml of 1% acetic acid. All samples were spiked with (18)O(4) isotope-labelled perchlorate internal standard prior to extraction. After purification on a graphitised carbon solid-phase extraction column, the extracts were injected into an ion chromatography system equipped with an Ionpac AS20 column for separation of perchlorate from other anions. The presence of perchlorate in samples was quantified by isotope dilution mass spectrometry. Analysis of both perchlorate and its isotope-labelled internal standard was carried out on a Waters Quattro Ultima triple quadrupole mass spectrometer operating in a multiple reaction monitoring (MRM) negative ionisation mode. The method was validated for linearity and range, accuracy, precision, sensitivity, and matrix effects. The limit of quantification (LOQ) was 0.4 ug l(-1) for liquid infant formula and 0.95 ug kg(-1) for powdered infant formula. The recovery ranged from 94% to 110% with an average of 98%. This method was used to analyse 39 infant formula, and perchlorate concentrations ranging from 2)-beta-d-glucopyranosyl-(1 -> 2)-beta-d-glu-copyranoside, based on the detailed analyses of the 1D and 2D NMR spectral data and acidic hydrolysis. Notoginsenoside ST-4 was investigated for its antiviral activity on herpes simplex type 1 (HSV-1) and type 2 (HSV-2) in vitro. The 50% effective concentration (EC(50)) values, determined by plaque reduction assay, were 16.47 +/- 0.67 and 19.44 +/- 1.16 MUM for HSV-1 and HSV-2, respectively, whereas the 50% cytotoxic concentration (CC(50)) determined by the XTT test on Vero cells was 510.64 +/- 4.56 MUM. As analyzed by attachment assay and penetration assay based on plaque reduction assay, the antiviral activity of notoginsenoside ST-4 was principally due to the penetration inhibition effects, which was confirmed by fluorescence microscopy observation that notoginsenoside ST-4 blocked the penetration of virus. Therefore, notoginsenoside ST-4 might be a promising agent for herpes simplex virus infection. PMID- 21623513 TI - Synthesis of 5-chloro-8-hydroxy-6-methoxy-3-pentylisocoumarin. AB - The synthesis of title isocoumarin, the 5-chloro analog of naturally occurring 7 chloro-8-hydroxy-6-methoxy-3-pentylisocoumarin, isolated from Tessmannia densiflora is described. Chlorination of ethyl 2-(2-ethoxy-2-oxoethyl)-4,6 dimethoxybenzoate (2) afforded 3-chloro ester (3) followed by hydrolysis to furnish the 2-(carboxymethyl)-3-chloro-4,6-dimethoxybenzoic acid (4) that was converted to corresponding anhydride (5). Condensation of the latter with hexanoyl chloride in the presence of tetramethylguanidine and triethyl amine afforded 5-chloro-6,8-dimethoxy-3-pentylisocoumarin (6) which upon regioselective demethylation yielded the title isocoumarin (1). PMID- 21623514 TI - Inhibition activities of polysaccharide (RG4-1) from Gentiana rigescens against RSV. AB - Respiratory syncytial virus (RSV) is the most important cause of lower respiratory tract infection in infants and young children. With the emergence of drug-resistant strains of RSV, new antiviral agents are needed urgently. Gentiana rigescens is a kind of Chinese herb, belonging to Gentianaceae, which has long been used as a folk medicine for curing inflammation, bacterial infection, viral infection, and so on. In this research, polysaccharide designated RG4-1 was isolated from G. rigescens by hot water extraction, ethanol precipitation, and macroreticular adsorbing resin column chromatography, and its antiviral activity, cytotoxicity, and possible antiviral mechanisms were assayed by cytopathogenic effect inhibition assay, 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, and plaque reduction assay. RG4-1 was a fructose-binding lectin. In host cell cultures, RG4-1 was found to be an effective antiviral component against RSV. It showed good inhibitory effect against RSV when it was added 2 h after virus infection with 50% effective concentration of 12.86 MUg/ml. RG4-1 also displayed its direct inactivation, attachment inhibition effect, and penetration inhibition effect against RSV. A time-dependent experiment was set up to confirm that RG4-1 blocked RSV infection at early stages of the infection. But RG4-1 seemed to be ineffective against intracellular virus and viral biosynthesis. PMID- 21623515 TI - Two new morphinane alkaloids from Sinomenium acutum. AB - Two new morphinane alkaloids, 1-hydroxy-10-oxo-sinomenine (1) and 4,5-epoxy-14 hydroxy sinomenine N-oxide (2), have been isolated from the stems of Sinomenium acutum. Their structures were established by various spectral analyses, especially 2D NMR experiments. The structure of 2 was confirmed by single crystal X-ray diffraction. The absolute configurations of 1 and 2 were deduced by comparison of CD spectra with the known alkaloid sinomenine (3). Compound 1 was tested for DPPH inhibition and gave IC(50) of 27.9 MUM. Compound 2 was tested for neuroprotective effect and showed significant activity against beta-amyloid(25 35)-induced oxidative injury (*P < 0.05) at 10 MUM in PC-12 cells. PMID- 21623516 TI - Three new sulfated triterpenoids from the roots of Gypsophila pacifica. AB - Three new sulfated triterpenoids (1-3), along with one known compound (4), were isolated from the roots of Gypsophila pacifica Kom. The structures of the new compounds were established as 3beta-O-sulfate gypsogenin 28-O-beta-d glucopyranosyl ester (1), 3beta-O-sulfate gypsogenin (2), and 3beta-O-sulfate quillaic acid (3) on the basis of 1D, 2D NMR, and HR-ESI-MS methods. PMID- 21623517 TI - Beneficial effects of cordycepin on metabolic profiles of liver and plasma from hyperlipidemic hamsters. AB - In this study, (1)H NMR-based metabonomics was applied to evaluate the beneficial effects of cordycepin (3'-deoxyadenosine), a natural monomer compound, on endogenous metabolic profiles of liver and plasma from hyperlipidemic Syrian golden hamsters. Hyperlipidemia was successfully established in hamsters fed by a high-fat diet for 2 weeks. The hyperlipidemic hamsters were treated with an oral administration of simvastatin (2 mg kg(- 1)) or cordycepin (140 mg kg(- 1)) for consecutive 4 weeks. The metabolic profiles of plasma and intact liver tissues were established using (1)H NMR spectroscopy. The results showed higher contents of lipids (triglyceride and cholesterol), lactate, acetate, alanine, glutamine together with lower contents of choline-containing compounds (e.g. phosphocholine, phosphatidylcholine, and glycerophosphocholine), glucose, and glycogen in plasma and liver samples from hyperlipidemic hamsters than those in controls. Cordycepin afforded a little lipid-regulating activity on plasma but more beneficial effects on liver, implicating that cordycepin might have a protective effect on liver under fatty liver condition. PMID- 21623518 TI - Dorstenpictanone: a new bicyclic polyprenylated compound from Dorstenia picta. AB - One new bicyclic polyprenylated compound, dorstenpictanone (1), was isolated from Dorstenia picta. The structure of the new compound was elucidated by detailed spectroscopic analysis such as (1)H NMR, (13)C NMR, COSY, HMQC, HMBC, and HREIMS. The relative configuration of dorstenpictanone (1) was distinguished by comparative analysis of the NMR spectral data with known analogues together with the ROESY experiment. PMID- 21623519 TI - A new highly oxygenated nortriterpenoid from Schisandra chinensis. AB - A new highly oxygenated nortriterpenoid, 2beta-hydroxy-micrandilactone C (1), and three related known triterpenoids (2-4) have been isolated from the leaves and stems of Schisandra chinensis. The structure of the new compound was elucidated by means of NMR and MS spectroscopic analyses. PMID- 21623520 TI - Chemical constituents from the roots of Feroniella lucida. AB - A new furanocoumarin named lucidafuranocoumarin A (7) together with 13 known coumarins (1-6, 8-14) and four known alkaloids (15-18) was isolated from the roots of Feroniella lucida. Their structures were elucidated on the basis of spectroscopic analysis. Some of the isolates were evaluated for their biological activities, and compound 18 showed strong cytotoxicity against KB (IC(50) = 0.637 MUg/ml) and NCI-H187 (IC(50) = 0.094 MUg/ml) human cancer cell lines, antimalarial activity against Plasmodium falciparum (IC(50) = 0.336 MUg/ml), and antituberculosis activity against Mycobacterium tuberculosis (MIC = 6.25 MUg/ml). PMID- 21623521 TI - A new drimane sesquiterpenoid glycoside from the seeds of Antiaris toxicaria. AB - A new drimane sesquiterpenoid glycoside, named 7-drimen-3beta,11-diol 3-O-beta-d glucopyranoside, was isolated from the 95% EtOH extract of the seeds of Antiaris toxicaria (Pers.) Lesch. The chemical structure was completely elucidated using a combination of 1D and 2D NMR techniques (COSY, HMQC, HMBC, and ROESY) and HR-ESI MS analysis. The compound showed inhibitory activities toward methicillin resistant Staphylococcus aureus (MRSA), chronic myelogenous leukemia (K562), and human hepatoma (SMMC-7721) cell lines. PMID- 21623522 TI - Cichorin A: a new benzo-isochromene from Cichorium intybus. AB - One new benzo-isochromene, named cichorin A (1), together with three known compounds oleanolic acid, beta-sitosterol, and beta-sitosterol glucopyranoside, was isolated from Cichorium intybus. The structure of the new compound was elucidated by detailed spectroscopic analysis such as (1)H, (13)C NMR, COSY, HMQC, HMBC, and HR-EI-MS. Relative configuration of asymmetric centers of cichorin A (1) was determined by the analysis of the (1)H NMR coupling constants together with the NOESY experiment. PMID- 21623523 TI - Sesquiterpenoids from Inula racemosa. AB - Phytochemical research on the roots of Inula racemosa yielded nine sesquiterpenoids including a new nor-eudesmane-type sesquiterpenoid, 11,12,13 trinoreudesm-5-en-7beta,8alpha-diol (1). The structures of isolated compounds were elucidated by extensive spectroscopic methods including 1D and 2D NMR. The structure of compound 2 was confirmed by single-crystal X-ray diffraction analysis. PMID- 21623524 TI - Phenolic compounds from Eurycorymbus cavaleriei. AB - Three new phenolic compounds, eurycorymboside A (1), eurycorymboside B (6), and eurycorymbic acid (8), were isolated from the stem part of Eurycorymbus cavaleriei (Sapindaceae) along with five known phenolic compounds, glucosyringic acid (2), vanillic acid 4-O-beta-d-glucoside (3), koaburaside (4), tachioside (5), and 4-hydroxy-3,5-bis(3-methyl-2-butenyl)benzaldehyde (7). The structures were established on the basis of spectral analysis. The antioxidant activities of compounds 1-6 were evaluated by the 1,1-diphenyl-2-picrylhydrazyl-free radical scavenging assay. Compound 4 exhibited antioxidant activity with an IC(50) value of 9.0 MUM. Compound 4 also showed weak inhibitory activity against influenza A neuraminidase. PMID- 21623525 TI - Constitutionality of the Patient Protection and Affordable Care Act under the Commerce Clause and the Necessary and Proper Clause. PMID- 21623526 TI - Recovery of damages for wrongful birth. PMID- 21623527 TI - Devouring childhood obesity by helping children help themselves. PMID- 21623528 TI - What did you say? Assisting hearing-impaired patients in hospitals. PMID- 21623531 TI - Vehicle design influences whole body vibration exposures: effect of the location of the front axle relative to the cab. AB - Using a repeated measure design, this study compared differences in whole body vibration (WBV) exposures among 13 drivers who drove a truck with the cab over the front axle (cab-over design) and a truck with the cab situated behind the front axle (non-cab-over design). The drivers drove both trucks over a standardized route that comprised three distinct segments: a freeway segment, a city street segment with stop-and-go driving (traffic lights), and a city street segment without traffic lights. A portable WBV data acquisition system collected tri-axial time-weighted and raw WBV data per ISO 2631-1 and 2631-5 standards. Simultaneous global positioning system (GPS) data were also collected to compare vehicle speeds. The GPS data indicated that there were no speed differences between the two vehicles. However, average and impulsive z-axis vibration levels were significantly higher for the cab-over design than for the non-cab-over design. In addition, significant WBV exposure differences between road types were found, with the freeway segments having the lowest exposures and the city street segments without traffic lights having the highest exposures. Vehicle type and the associated WBV exposures should be considered when purchasing vehicles to be used by full-time professional vehicle operators. PMID- 21623532 TI - Survey results of the training, nutrition, and mental preparation of triathletes: practical implications of findings. AB - Although triathlon is growing in popularity at a remarkable rate, it has not been extensively studied. The aims of this research were to identify preparation strategies used by triathletes and to categorize these strategies according to gender and consultation with triathlon coaches. Survey data collected from 401 triathletes (207 males, 194 females) revealed training, nutritional, and mental preparation habits. Most participants engaged in strength training, consumed food and/or fluids during and after training, set training and competition goals, and applied mental preparation strategies during training and the hour before racing. Water was the most commonly consumed fluid; positive self-talk was the most used mental strategy. Participants were more likely to consult with a triathlon coach than a nutrition or sport psychology professional. Athletes with more years of experience in triathlon and those competing in longer distances were more likely to consult a triathlon coach. Female triathletes were more likely than male triathletes to train with others, use mental preparation strategies, and report feeling anxious before competitions. More male triathletes reported using nutritional supplements during training than their female counterparts. These findings add to the limited research base on triathletes' training habits, and hopefully will help guide practitioners who work with this group. The results provide guidance for collaborative efforts among training, nutrition, and mental health professionals to best support triathletes. PMID- 21623533 TI - Benefits of achieving vigorous as well as moderate physical activity recommendations: evidence from heart rate complexity and cardiac vagal modulation. AB - The aim of this study was to examine differences in traditional heart rate variability measurements and heart rate complexity (sample entropy) in young adults grouped by objectively measured achievement of either moderate or both moderate and vigorous physical activity recommendations. Of 168 young adults tested (86 females, 82 males; age 20.5 +/- 1.2 years), 119 achieved only recommendations for moderate physical activity (moderate group) and 49 achieved recommendations for both moderate and vigorous physical activity (vigorous group). Analysis of covariance controlling for sex, weekly minutes of moderate physical activity, and percentage of body fat was used to assess between-group differences in heart rate variability and heart rate complexity. Logistic regression analysis was used to determine the group characteristics that best predicted high heart rate complexity and vagal indices of heart rate variability. The majority of the autonomic measures were higher (P < 0.05) in the vigorous group, and regression analysis showed that vigorous physical activity was the only multivariate predictor of higher heart rate complexity and higher heart rate variability. Young adults engaged in regular vigorous physical activity were more than twice as likely to have high heart rate complexity than those involved in predominantly moderate exercise. These findings suggest that vigorous physical activity is more closely associated with high heart rate complexity than moderate physical activity in young adults. PMID- 21623534 TI - Interactive effect of cigarette smoke extract and world trade center dust particles on airway cell cytotoxicity. AB - Rescue workers and residents exposed to the environment surrounding the collapse of the World Trade Center (WTC) on September 11, 2001, have suffered a disproportionate incidence of chronic lung disease attributed to the inhalation of airborne dust. To date, the pathophysiology of this lung disease is poorly understood. The aim of this study was to examine whether airborne dust contaminants recovered from the surrounding area 24-48 h after the collapse of the WTC demonstrate direct cytotoxicity to two airway cell types that were most directly exposed to inhaled dust, airway epithelial and smooth muscle cells. It was also of interest to determine whether the presence of these dusts could modulate the effects of cigarette smoke on these cell types in that some of the individuals who responded to the collapse site were also smokers. Human cultured airway epithelial (BEAS-2B) cells were exposed to 10% cigarette smoke extract (CSE), WTC dust particles (10-53 MUm; 0.01-0.5 MUg/MUl), or a combination of the two for 2-24 h. Cell viability was measured by determining mitochondrial integrity (MTT assays) and apoptosis (poly-ADP-ribose polymerase [PARP] immunoblotting). Conditioned cell culture media recovered from the CSE- and/or WTC dust-exposed BEAS-2B cells were then applied to cultured human airway smooth muscle cells that were subsequently assayed for mitochondrial integrity and their ability to synthesize cyclic AMP (a regulator of airway smooth muscle constriction). BEAS-2B cells underwent necrotic cell death following exposure to WTC dust or CSE for 2-24 h without evidence of apoptosis. Smooth muscle cells demonstrated cellular toxicity and enhanced cyclic AMP synthesis following exposure to conditioned media from WTC- or CSE-exposed epithelial cells. These acute toxicity assays of WTC dust and CSE offer insights into lung cell toxicity that may contribute to the pathophysiology of chronic lung disease in workers and residents exposed to WTC dust. These studies clearly showed that WTC dust (at least the supercoarse particle fraction) or CSE alone exerted direct adverse effects on airway epithelial and smooth muscle cells, and altered the signaling properties of airway smooth muscle cells. In addition the combination of CSE and WTC exerted an interactive effect on cell toxicity. It remains to be determined whether these initial cell death events might account, in part, for the chronic lung effects associated with WTC dust exposure among First Responders and others. PMID- 21623535 TI - A positive association found between autism prevalence and childhood vaccination uptake across the U.S. population. AB - The reason for the rapid rise of autism in the United States that began in the 1990s is a mystery. Although individuals probably have a genetic predisposition to develop autism, researchers suspect that one or more environmental triggers are also needed. One of those triggers might be the battery of vaccinations that young children receive. Using regression analysis and controlling for family income and ethnicity, the relationship between the proportion of children who received the recommended vaccines by age 2 years and the prevalence of autism (AUT) or speech or language impairment (SLI) in each U.S. state from 2001 and 2007 was determined. A positive and statistically significant relationship was found: The higher the proportion of children receiving recommended vaccinations, the higher was the prevalence of AUT or SLI. A 1% increase in vaccination was associated with an additional 680 children having AUT or SLI. Neither parental behavior nor access to care affected the results, since vaccination proportions were not significantly related (statistically) to any other disability or to the number of pediatricians in a U.S. state. The results suggest that although mercury has been removed from many vaccines, other culprits may link vaccines to autism. Further study into the relationship between vaccines and autism is warranted. PMID- 21623536 TI - The effects of perchlorate on thyroidal gene expression are different from the effects of iodide deficiency. AB - Perchlorate (ClO4-), which is a ubiquitous and persistent ion, competitively interferes with iodide (I) accumulation in the thyroid, producing I deficiency (ID), which may result in reduced thyroid hormone synthesis and secretion. Human studies suggest that ClO4- presents little risk in healthy individuals; however, the precautionary principle demands that the sensitive populations of ID adults and mothers require extra consideration. In an attempt to determine whether the effects on gene expression were similar, the thyroidal effects of ClO4- (10 mg/kg) treatment for 14 d in drinking water were compared with those produced by 8 wk of ID in rats. The thyroids were collected (n = 3 each group) and total mRNA was analyzed using the Affymetrix Rat Genome 230 2.0 GeneChip. Changes in gene expression were compared with appropriate control groups. The twofold gene changes due to ID were compared with alterations due to ClO4- treatment. One hundred and eighty-nine transcripts were changed by the ID diet and 722 transcripts were altered by the ClO4- treatment. Thirty-four percent of the transcripts changed by the I-deficient diet were also altered by ClO4- and generally in the same direction. Three specific transporter genes, AQP1, NIS, and SLC22A3, were changed by both treatments, indicating that the membrane-specific changes were similar. Iodide deficiency primarily produced alterations in retinol and calcium signaling pathways and ClO4- primarily produced changes related to the accumulation of extracellular matrix proteins. This study provides evidence that ClO4-, at least at this dose level, changes more genes and alters different genes compared to ID. PMID- 21623537 TI - Biomarker measurements of concurrent exposure to multiple environmental chemicals and chemical classes in children. AB - Concern is mounting that children from disadvantaged, low-income neighborhoods are likely to be both more exposed to chemical hazards and more susceptible to related adverse health effects. This article reports measurements of >75 individual biomarkers spanning 7 chemical/pollutant classes in blood and urine from more than 100 children living in a socioeconomically disadvantaged and ethnically diverse area of south Minneapolis, MN. Results indicate that a significant proportion of children in the study were at the high end of the exposure distribution compared to national reference ranges for a variety of environmental chemicals and/or their metabolites, including phthalates, organochlorine pesticides, organophosphate pesticides, metals, polychlorinated biphenyls, and volatile organic compounds. In addition, levels of cotinine in urine indicate that more than half the children were regularly exposed to environmental tobacco smoke, with the upper 10th percentile exposed to relatively high concentrations. PMID- 21623538 TI - The biocontrol fungus Trichoderma stromaticum downregulates respiratory burst and nitric oxide in phagocytes and IFN-gamma and IL-10. AB - Trichoderma stromaticum, a biocontrol agent of the cacao witches' broom pathogen Moniliophthora perniciosa, has been used in Brazil as part of the integrated pest management of cacao. At the present time, little is known about the effects of T. stromaticum on the modulation of in vitro or in vivo immune responses. The present study examined the interaction of T. stromaticum spores with cellular and molecular components of the immune system following intranasal sensitization of mice. Our results showed that T. stromaticum spores prevented the expression and production of inflammatory mediators in macrophages stimulated with interferon (IFN)-gamma plus lipopolysaccharide (LPS) and neutrophils stimulated with phorbol myristate 13-acetate (PMA). Quantitative polymerase chain reaction (qPCR) assays revealed that T. stromaticum spores inhibited the expression of dectin-1 and Toll like-receptor (TLR)2/TLR4. Intranasal injection of BALB/c mice and subsequent challenge with spores of T. stromaticum induced a discrete inflammatory response in the lungs. Interestingly, the spores inhibited local and systemic production of the regulatory IL-10 and proinflammatory IFN-gamma cytokines. In addition the spores presented an antiproliferative effect on spleen cells. These findings showed that the biopesticide T. stromaticum may exert immunosuppressive effects in vitro and in vivo. PMID- 21623539 TI - Peptide nucleic acid Pt(II) conjugates: a preliminary study of antisense effects in Xenopus laevis. AB - The avid hybridization of peptide nucleic acid (PNA) to DNA and RNA, coupled with the analogue's stability toward enzymatic degradation, has led to its investigation as an antigene/antisense agent. PNA targeted toward the 5'-UTR of an mRNA transcript can effect efficient silencing; however, if targeted to an area within the coding region, the PNA can be displaced by the moving ribosome and be an ineffective antisense agent. Platinum-appended and standard PNAs antisense to an area within the open-reading frame of the gene noggin, were injected into Xenopus laevis embryos. Phenotypic responses were observed and the preliminary results are reported herein. PMID- 21623540 TI - Synthesis and anti-HIV evaluation of 3'-triazolo nucleosides. AB - A series of hitherto unknown 3'-alpha-[1,2,3]-substituted triazolo-2',3' dideoxypyrimidine nucleoside analogues of the anti-HIV 3'-azido-3'-deoxythymidine (AZT) were synthesized through catalyzed alkyne-azide 1,3-dipolar cycloaddition (Huisgen reaction). Those 3'-[1,2,3]-triazolo analogues bearing an azido alkyl chain were evaluated for their anti-HIV activity against HIV-1 in primary human lymphocytes as well as for their cytotoxicity in different cells. None of them inhibit HIV replication (EC(50) > 20 MUM); two of them were converted to their triphosphate form to evaluate their HIV-RT inhibition. PMID- 21623541 TI - Mild detritylation of nucleic acid hydroxyl groups by warming up. AB - It is challenging to effectively deprotect hydroxyl groups of acid-or-base sensitive bio-macromolecules without causing even minor defects and compromising high quality of final products. We report here a mild detritylation strategy in mildly acidic buffers to remove the DMTr protection from the 5'-hydroxyl groups of synthetic nucleic acids. The DMTr-groups can be easily and effectively removed at pH 4.5 or 5.0 with slight warming up (40 degrees C), offering virtually quantitative deprotection. This warming-up strategy is particularly useful for deprotection of the modified nucleic acids that are sensitive to the conventional acid deprotection. As a first step towards our long-term goal of synthesizing defect-free nucleic acids, our novel and simple strategy further increases the quality of synthetic nucleic acids. PMID- 21623542 TI - Synthesis and insecticidal activities of novel 1,3,4-thiadiazole 5-fluorouracil acetamides derivatives: an RNA interference insecticide. AB - A series of novel 1,3,4-thiadiazole 5-fluorouracil acetamides derivatives were designed and synthesized. Their structures were confirmed by infrared, (1)H NMR spectroscopy, and elemental analysis. The insecticidal activities against Tetranychus cinnabarinus and Aphis craccivora of these new compounds were evaluated. The bioassay tests showed that most of these title compounds possessed a good combination of stomach toxicity as well as contact toxicity against Tetranychus cinnabarinus and Aphis craccivora. In particular, the insecticidal activity of the title compound IVe against Aphis craccivora was better than the commercialized thiacloprid and was also comparable to another commercialized product, imidacloprid. The introduction of fluorines to meta and para-position of the benzene ring was essential for high bioactivity. PMID- 21623543 TI - Synthesis, crystal structure, and in vitro biological evaluation of C-6 pyrimidine derivatives: new lead structures for monitoring gene expression in vivo. AB - Novel C-6 substituted pyrimidine derivatives are good substrates of herpes simplex virus type 1 thymidine kinase (HSV1-TK). Enzyme kinetic experiments showed that our lead compound, N-methyl DHBT (N-methyl-6-(1,3-dihydroxyisobutyl) thymine; N-Me DHBT), is phosphorylated at a similar rate compared to "gold standard" 9-[4-fluoro-3-(hydroxymethyl)butyl]guanine, FHBG, (K(m) = 10 +/- 0.3 MUM; k(cat) = 0.036 +/- 0.015 sec(-1)). Additionally, it does not show cytotoxic properties on B16F1 cells up to a concentration of 10 mM. The x-ray analysis of the crystal structures of HSV1-TK with N-Me DHBT and of HSV1-TK with the fluorinated derivative N-Me FHBT confirmed the binding mode predicted by docking studies and their substrate characteristics. Moreover, the crystal structure of HSV1-TK with N-Me DHBT revealed an additional water-mediated H-bond interesting for the design of further analogues. PMID- 21623544 TI - Perceived stress in prodromal Huntington disease. AB - This study examines perceived stress and its relationship to depressive symptoms, life changes and functional capacity in a large sample of individuals who are positive for the Huntington disease (HD) gene expansion but not yet diagnosed. Participants were classified by estimated proximity to HD diagnosis (far, mid, near) and compared with a non-gene-expanded comparison group. Persons in the mid group had the highest stress scores. A significant interaction between age and time since HD genetic testing was also found. Secondary analyses using data from a different data collection point and including a diagnosed group showed the highest stress scores in the diagnosed group. Possible explanations and implications are discussed. PMID- 21623545 TI - Relations between acceptance of multiple sclerosis and positive and negative adjustments. AB - This study examines relations between acceptance as defined in Acceptance and Commitment Therapy (ACT) and adjustment to multiple sclerosis (MS). A first step in this investigation was the development of a measure of acceptance of MS called the MS Acceptance Questionnaire (MSAQ). Consistent with prior findings and theoretical propositions, it was predicted that acceptance would be associated with better adjustment to MS (lower distress and higher positive affect, life satisfaction and marital adjustment and better health). A total of 128 persons with MS completed measures of demographics, illness and adjustment at Time 1 and measures of acceptance and adjustment 12 months later (Time 2). Factor analyses of the MSAQ revealed two factors, action and willingness. Associations between the MSAQ and other validated acceptance measures supported convergent validity. As predicted, after controlling for the effects of initial adjustment and relevant demographic and illness variables, greater acceptance was related to better adjustment, although the action factor emerged as the strongest predictor of better adjustment. This is the first study to examine the role of acceptance (as defined in ACT) in adjusting to MS over time and as such provides a first step for further investigation of acceptance in MS. PMID- 21623546 TI - Affect integration and reflective function: clarification of central conceptual issues. AB - The importance of affect regulation, modulation or integration for higher-order reflection and adequate functioning is increasingly emphasized across different therapeutic approaches and theories of change. These processes are probably central to any psychotherapeutic endeavor, whether explicitly conceptualized or not, and in recent years a number of therapeutic approaches have been developed that explicitly target them as a primary area of change. However, there still is important lack of clarity in the field regarding the understanding and operationalization of affect integration, particularly when it comes to specifying underlying mechanisms, the significance of different affect states, and the establishment of operational criteria for measurement. The conceptual relationship between affect integration and reflective function thus remains ambiguous. The present article addresses these topics, indicating ways in which a more complex and exhaustive understanding of integration of affect, cognition and behavior can be attained. PMID- 21623547 TI - Changes in self and object representations following psychotherapy measured by a theory-free, computational, semantic space method. AB - We propose a theory-neutral, computational and data-driven method for assessing changes in semantic content of object representations following long-term psychodynamic psychotherapy. Young adults in psychotherapy are compared with an age-matched, non-clinical sample at three time points. Verbatim transcripts of descriptions of the self and parents were quantified in a semantic space constructed by Latent Semantic Analysis. In the psychotherapy group, all representations changed from baseline to follow-up, whereas no comparable changes could be observed in the comparison group. The semantic space method supports the hypothesis that long-term psychodynamic psychotherapy contributes to sustained change of affective-cognitive schemas of self and others. PMID- 21623548 TI - Corrective interpersonal experience in psychodrama group therapy: a comprehensive process analysis of significant therapeutic events. AB - This study investigated the process of resolving painful emotional experience during psychodrama group therapy, by examining significant therapeutic events within seven psychodrama enactments. A comprehensive process analysis of four resolved and three not-resolved cases identified five meta-processes which were linked to in-session resolution. One was a readiness to engage in the therapeutic process, which was influenced by client characteristics and the client's experience of the group; and four were therapeutic events: (1) re-experiencing with insight; (2) activating resourcefulness; (3) social atom repair with emotional release; and (4) integration. A corrective interpersonal experience (social atom repair) healed the sense of fragmentation and interpersonal disconnection associated with unresolved emotional pain, and emotional release was therapeutically helpful when located within the enactment of this new role relationship. Protagonists who experienced resolution reported important improvements in interpersonal functioning and sense of self which they attributed to this experience. PMID- 21623549 TI - The impact of alliance ruptures on client outcome expectations in cognitive behavioral therapy. AB - The present study examined the impact of alliance ruptures (AR) on outcome expectations among 38 individuals who received cognitive-behavioral therapy for generalized anxiety disorder. ARs had a significant adverse impact on immediate post-rupture outcome expectations compared to dyads experiencing no AR. ARs also had a far greater negative impact on outcome expectations among those who were initially skeptical about treatment. Finally, path analysis indicated that the impact of an AR on posttreatment outcome was mediated by its influence on outcome expectations. These findings suggest an important role for preventing and repairing ARs, particularly among those who initially have doubts about treatment benefit. PMID- 21623550 TI - Therapist effectiveness: implications for accountability and patient care. AB - Significant therapist variability has been demonstrated in both psychotherapy outcomes and process (e.g., the working alliance). In an attempt to provide prevalence estimates of "effective" and "harmful" therapists, the outcomes of 6960 patients seen by 696 therapists in the context of naturalistic treatment were analyzed across multiple symptom and functioning domains. Therapists were defined based on whether their average client reliably improved, worsened, or neither improved nor worsened. Results varied by domain with the widespread pervasiveness of unclassifiable/ineffective and harmful therapists ranging from 33 to 65%. Harmful therapists demonstrated large, negative treatment effect sizes (d= -0.91 to -1.49) while effective therapists demonstrated large, positive treatment effect sizes (d=1.00 to 1.52). Therapist domain-specific effectiveness correlated poorly across domains, suggesting that therapist competencies may be domain or disorder specific, rather than reflecting a core attribute or underlying therapeutic skill construct. Public policy and clinical implications of these findings are discussed, including the importance of integrating benchmarked outcome measurement into both routine care and training. PMID- 21623551 TI - Therapeutic work with the present moment: a comparative conversation analysis of existential and cognitive therapies. AB - Therapeutic work with the client's present moment experience in existential therapy was studied by means of conversation analysis. Using publicly available video recordings of therapy sessions as data, an existential therapist's practice of guiding a client into immediacy, or refocusing the talk on a client's immediate experience, was described and compared with a therapist's corresponding action in cognitive therapy. The study contributes to the description of interactional practice of existential therapy, and involves the first application of conversation analysis to a comparative study of psychotherapy process. The potential utility of this approach and the clinical and empirical implications of the present findings are discussed. PMID- 21623552 TI - Therapeutic change in interaction: conversation analysis of a transforming sequence. AB - A process of change within a single case of cognitive-constructivist therapy is analyzed by means of conversation analysis (CA). The focus is on a process of change in the sequences of interaction, which consist of the therapist's conclusion and the patient's response to it. In the conclusions, the therapist investigates and challenges the patient's tendency to transform her feelings of disappointment and anger into self-blame. Over the course of the therapy, the patient's responses to these conclusions are recast: from the patient first rejecting the conclusion, to then being ambivalent, and finally to agreeing with the therapist. On the basis of this case study, we suggest that an analysis that focuses on sequences of talk that are interactionally similar offers a sensitive method to investigate the manifestation of therapeutic change. It is suggested that this line of research can complement assimilation analysis and other methods of analyzing changes in a client's talk. PMID- 21623553 TI - Safety at play: let's all help to prevent drowning. PMID- 21623554 TI - Are injuries correlated?: interpreting the intraclass correlation coefficient. PMID- 21623555 TI - Long-term results of salvage photodynamic therapy for patients with local failure after chemoradiotherapy for esophageal squamous cell carcinoma. AB - BACKGROUND AND STUDY AIMS: Local failure after chemoradiotherapy (CRT) remains a major problem for patients with esophageal squamous cell carcinoma (ESCC). The aim of this study was to clarify the long-term results of salvage photodynamic therapy (PDT) for local failure. PATIENTS AND METHODS: Patients were treated with CRT, consisting of more than 50 Gy irradiation and concurrent chemotherapy. The indications for salvage PDT were as follows: 1) absence of lymph-node or distant metastasis after CRT; 2) failure lesion limited to T2; 3) refusal by patient to undergo salvage esophagectomy; 4) written informed consent. PDT was performed using an excimer dye laser at 48 and 72 hours after administration of Photofrin. RESULTS: A total of 37 consecutive patients underwent salvage PDT. The baseline stage before CRT was as follows: T1/T2/T3/T4 in 3/4/24/6 and N0/1 in 13/24 patients, respectively. Prior to PDT, 20 patients had a uT1 lesion, and 17 had a uT2 lesion; 24 patients had histologically proven local failure. A complete response was achieved in 22 patients (59.5%) following PDT. Esophageal fistulae, stenosis, and phototoxicity occurred in 4 (10.8%), 20 (54.1%), and 2 (5.4%) patients, respectively. Over a median follow-up period of 55 months, the 5-year progression-free (PFS) and overall survival rates of 37 patients following PDT were 20.7% and 36.1%, respectively. The 5-year PFS and overall survival of 24 patients with proven local failure were 17.6% and 34.6%, respectively. CONCLUSION: Salvage PDT is a curative treatment option for patients with local failure after CRT for ESCC. PMID- 21623556 TI - A quantitative assessment of the risks and cost savings of forgoing histologic examination of diminutive polyps. AB - BACKGROUND AND AIMS: Endoscopic prediction of polyp histology is rapidly improving to the point where it may not be necessary to submit all polyps for formal histologic assessment. This study aimed to quantify the expected costs and outcomes of removing diminutive polyps without subsequent pathologic assessment. METHODS: Cross-sectional analysis of a colonoscopy database for polyp histology; decision models that quantify effects on guideline-recommended surveillance and subsequent costs and consequences. The database was composed of consecutive colonoscopies from 1999 to 2004 at a single-institution tertiary care center. Patients were those found to have at least one diminutive polyp removed during colonoscopy, irrespective of indication. The main outcome measurements include up front cost savings resulting from forgoing pathologic assessment; frequency and cost of incorrect surveillance intervals based on errors in histologic assessment; number needed to harm (NNH) for perforation and/or interval cancer. RESULTS: Incorrect surveillance intervals were recommended in 1.9% of cases when tissue was submitted for pathologic assessment and 11.8% of cases when it was not. Based on the annual volume of colonoscopy in the US, the annual up-front cost savings of forgoing the pathologic assessment would exceed a billion dollars. An upper estimate on the downstream costs and consequences of forgoing pathology suggests that less than 10% of the up-front savings would be offset and the NNH exceeds 11000. CONCLUSION: Endoscopic diagnosis of polyp histology during colonoscopy and forgoing pathologic examination would result in substantial up front cost savings. Downstream consequences of the resulting incorrect surveillance intervals appear to be negligible. PMID- 21623557 TI - Cost effectiveness and projected national impact of colorectal cancer screening in France. AB - BACKGROUND: Colorectal cancer (CRC) is a major cause of morbidity and mortality in France. Only scanty data on cost-effectiveness of CRC screening in Europe are available, generating uncertainty over its efficiency. Although immunochemical fecal tests (FIT) and guaiac-based fecal occult blood tests (g-FOBT) have been shown to be cost-effective in France, cost-effectiveness of endoscopic screening has not yet been addressed. METHODS: Cost-effectiveness of screening strategies using colonoscopy, flexible sigmoidoscopy, second-generation colon capsule endoscopy (CCE), FIT and g-FOBT were compared using a Markov model. A 40 % adherence rate was assumed for all strategies. Colonoscopy costs included anesthesiologist assistance. Incremental cost-effectiveness ratios (ICERs) were calculated. Probabilistic and value-of-information analyses were used to estimate the expected benefit of future research. A third-payer perspective was adopted. RESULTS: In the reference case analysis, FIT repeated every year was the most cost-effective strategy, with an ICER of ?48165 per life-year gained vs. FIT every 2 years, which was the next most cost-effective strategy. Although CCE every 5 years was as effective as FIT 1-year, it was not a cost-effective alternative. Colonoscopy repeated every 10 years was substantially more costly, and slightly less effective than FIT 1-year. When projecting the model outputs onto the French population, the least (g-FOBT 2-years) and most (FIT 1-year) effective strategies reduced the absolute number of annual CRC deaths from 16037 to 12916 and 11217, respectively, resulting in an annual additional cost of ?26 million and ?347 million, respectively. Probabilistic sensitivity analysis demonstrated that FIT 1-year was the optimal choice in 20% of the simulated scenarios, whereas sigmoidoscopy 5-years, colonoscopy, and FIT 2-years were the optimal choices in 40%, 26%, and 14%, respectively. CONCLUSIONS: A screening program based on FIT 1-year appeared to be the most cost-effective approach for CRC screening in France. However, a substantial uncertainty over this choice is still present. PMID- 21623558 TI - Effectiveness of a novel endoscopy training concept. AB - BACKGROUND AND STUDY AIMS: Training standards in gastrointestinal endoscopy are poorly defined even though different simulators are increasingly used for skills training. In 2001 a new training concept called "GATE--gastroenterological education-training endoscopy" was established, which provides a combination of background theory, video demonstrations, and simulator training. We aimed to evaluate the acceptance and training effect of this training model. METHODS: In total, 98 physicians participating in four training courses were included. Data were collected on baseline characteristics, acceptance (5-point Likert scale), and pre- and post-course knowledge through a structured questionnaire (A-type and Pick-N multiple choice questions). A total of 13 trainees were randomly selected for additional simulator assessment of training effects on manual skills (5-point Likert scale). RESULTS: A total of 78 trainees (80%) provided complete data sets. The evaluation showed a positive acceptance of the training program (value 1 and 2, Likert scale); for example, 88% of participants suggested the inclusion of the GATE course as an obligatory part of endoscopic education. There was a significant improvement in theoretical knowledge in the post-test set compared with the pre-test set (mean 3.27 +/-1.30 vs. 1.69 +/-1.01 points; P<0.001). The training effect on practical skill showed a significant reduction in time needed for a procedure (445 +/-189 s vs. 274 +/-129 s; P<0.01). The mean assessment rating for practical skills improved from 3.05 +/-0.65 at baseline to 2.52 +/ 0.59 on Likert scale ( P=0.085). CONCLUSIONS: The integrated GATE training improved theoretical knowledge and manual skill. The GATE courses have been accredited by the German Society of Gastroenterology, underlining the demand for implementing preclinical training courses in endoscopic training. PMID- 21623559 TI - Low dose endoluminal photodynamic therapy improves murine T cell-mediated colitis. AB - BACKGROUND AND STUDY AIMS: Low dose photodynamic therapy (LDPDT) may modify the mucosal immune response and may thus provide a therapy for Crohn's disease. We evaluated the efficacy and safety of this technique in a murine T cell-mediated colitis model. METHODS: The safety of LDPDT was first tested in BALB/c mice. Naive T cells were used to induce colitis in mice with severe combined immunodeficiency, which were followed up endoscopically, and a murine endoscopic index of colitis (MEIC) was developed. The efficacy of LDPDT (10 J/cm (2); delta aminolevulinic acid, 15 mg/kg bodyweight) was then tested on mice with moderate colitis, while a disease control group received no treatment. The MEIC, weight, length, and histology of the colon, cytokine expression indices, number of mucosal CD4 (+) T cells, percentage of apoptotic CD4 (+) T cells, body weight, and systemic side effects were evaluated. RESULTS: LDPDT improved the MEIC ( P = 0.011) and the histological score ( P = 0.025), diminished the expression indices of the proinflammatory cytokines, interleukin-6 ( P = 0.042), interleukin-17 ( P = 0.029), and interferon-gamma ( P = 0.014), decreased the number of mucosal CD4 (+) T cells, and increased the percentage of apoptotic CD4 (+) T cells compared with the disease control group. No local or systemic side effects occurred. CONCLUSION: LDPDT improves murine T cell-mediated colitis, decreases the proinflammatory cytokines interleukin-6, interleukin-17, and interferon-gamma, and decreases the number of CD4 (+) T cells. No adverse events were observed. Therefore, this technique is now being evaluated in patients with inflammatory bowel disease. PMID- 21623560 TI - Endoscopic submucosal dissection in a European setting. A multi-institutional report of a technique in development. AB - BACKGROUND AND STUDY AIMS: Endoscopic submucosal dissection (ESD) is a technique for "en bloc" resection of superficial tumors of the gastrointestinal tract. In France, experience with this technique is still limited. We wanted to assess the development of ESD in France, with special attention to short term outcomes. PATIENTS AND METHODS: Members of the Societe Francaise d'Endoscopie Digestive (SFED) who declared performing ESD reported their cases prospectively on a voluntary basis. Demographic, clinical, and technical data, and the results of immediate complications were collected. Case reports were completed prospectively by each investigator before pooled analysis. RESULTS: A total of 188 consecutive case reports were collected from 16 centers. The median case mix per center was 6 patients (range 1-43). The lesion sites treated by ESD were the stomach (n = 75), esophagus (n = 27), duodenum (n = 1), cecum (n = 2), right colon (n = 3), transverse colon (n = 5), sigmoid (n = 3), and rectum (n = 72). The median size of the lesions was 26 mm (range 2-150 mm). En bloc resection was achieved in 77.1% of cases, with complete R0 resection in 72.9%. Histopathology results showed high grade dysplasia or superficial cancer in 71.2%. The median duration of ESD was 105 minutes (range 20-450 minutes). The short term morbidity was 29.2% including 34 cases of perforation (18.1%), and 21 hemorrhages (11.2%) during the 24 hours following ESD, 89% of which were managed conservatively or endoscopically. CONCLUSION: In this early experience, the feasibility of ESD appeared to be good but R0 resection and complication rates did not match those reported by Japanese authors and must be improved by an extended practice. PMID- 21623561 TI - Defining the endoscopic appearances of tylosis using conventional and narrow-band imaging: a case series. AB - Tylosis is an autosomal dominant skin disorder strongly associated with esophageal squamous cell cancer. We present a single-operator experience of utilizing conventional endoscopy and narrow-band imaging with magnification to characterize esophageal appearances in tylosis. Nineteen consecutive patients with tylosis attending for surveillance endoscopy were studied. White-light imaging (WLI) and narrow-band imaging (NBI) were undertaken, with magnification being performed as necessary. On WLI, we classified 12 patients as having mild change, 5 moderate change, and 2 severe change. WLI can define changes to the esophageal mucosa of variable hyperkeratosis and identify more significant focal abnormalities. NBI enhances these mucosal changes, and NBI with magnification can demonstrate intrapapillary capillary loop changes compatible with dysplasia, prompting consideration of surgery. This report is the first to characterize the endoscopic appearances in tylosis. PMID- 21623562 TI - Biodegradable stents for the treatment of benign stenoses of the small and large intestines. AB - Biodegradable stents, which are made of various synthetic polymers, such as polylactide or polyglycolide, or co-polymers, such as polydioxanone, can be used for the treatment of benign refractory stenoses of the gastrointestinal tract. Here we report 11 patients (median age 41) with stenosing Crohn's disease of the small and/or large intestine. Endoscopic insertion of a biodegradable stent was successful at the first attempt in all patients except one. Subsequent follow-up was for a mean of 16 months, median 17 months, range 12-29 months. Early stent migration (between 2 days and 8 weeks) was seen in three patients. Mucosal overgrowth (epithelial hyperplasia) was not observed in any of the patients during the follow-up period. The high rate of early stent migration might be solved by appropriate tailoring and further improvements in the design of the biodegradable stents. Proof of long-term efficacy and safety requires further studies. PMID- 21623563 TI - Benjamin Alcock and the pudendal canal. AB - The anatomy of the pudendal nerve is complex and difficult to visualize. Entrapment of the pudendal nerve is believed to occur in a canal, the pudendal canal or Alcock's canal, yet in the literature this term is used to refer to several different anatomic locations. We present a brief history of Benjamin Alcock, and we compare Alcock's original description of the pudendal canal with our findings from a cadaveric study. It is concluded that Alcock's canal for the pudendal nerve, as Alcock described it related to the pudendal artery, should be that portion of the pudendal nerve within the obturator internus fascia. This definition now permits future medical and surgical approaches to use the appropriate terminology for this anatomic location. PMID- 21623564 TI - Calcaneal reconstruction with free fibular osteocutaneous flap. AB - Due to the role of the calcaneus in weight bearing, soft tissue coverage along with proper reduction of the fracture is the treatment following open calcaneal injury. Intra-articular calcaneal fractures present a very difficult management problem, as the lack of soft tissue and the intricate vascularity in this area pose a risk of complications. Coverage with local and free muscle flaps following excision of infected structures is a common approach for the treatment of chronic osteomyelitis. However, it is unknown which type of flap is optimal for the treatment of lateral foot wounds, especially when complicated by calcaneal osteomyelitis. A patient presented with an open wound over the lateral aspect of the heel with exposed hardware and chronic osteomyelitis of the calcaneus. Following multiple debridements, an ipsilateral osteocutaneous free fibular flap was transferred to the bony defect. Weight bearing was initiated at 2 months postoperatively, and he now ambulates with a normal gait, has normal plantar sensation, and has no difficulty maneuvering stairs. The patient has done well postoperatively and has recovered full range of motion and complete mobility. In this case report, an osteocutaneous free flap provided an excellent outcome for an active patient with a very complex and complicated condition. PMID- 21623568 TI - High tech versus old school: will traditional craftsmanship prevail? PMID- 21623570 TI - Ginkgolide B promotes proliferation and functional activities of bone marrow derived endothelial progenitor cells: involvement of Akt/eNOS and MAPK/p38 signaling pathways. AB - Bone marrow-derived, circulating endothelial progenitor cells (EPCs) contribute to neovascularization in various diseases, and represent a very interesting alternative cell source for enhancing vasculogenesis in regenerative medicine. In this study, we investigated the effects of Ginkgolide B (GB) on proliferation and differentiation of EPCs, and the involved signaling pathway in vitro. EPC proliferation, migration, adhesion and angiogenesis activities were assessed with the WST-8 assay, Transwell chamber assay, cell counting and angiogenesis kit, respectively. Apoptosis was detected with annexin V and propidium iodide staining. The protein expression of angiogenesis-related makers was detected by Western blot, and related gene expression was determined by real-time polymerase chain reaction (RT-PCR). The results showed that GB promoted the proliferation and endothelial gene expression, and markedly enhanced vascular endothelial growth factor-induced migration response and the capability to incorporate into the vascular networks in EPCs. GB protected EPCs from H2O2-induced cell death. GB induced the phosphorylation of eNOS, Akt and p38, which in turn promoted cell proliferation and function. In conclusion, the present study demonstrates that GB, at a near medical applied dose, increases the number and functional activities of EPCs with involvement of Akt/endothelial nitric oxide synthase and mitogen-activated protein kinase (MAPK)/p38 signal pathways. These findings raise the intriguing possibility that GB may play an important role in the protection and revascularization of blood vessels. PMID- 21623571 TI - Cartilage damage involving extrusion of mineralisable matrix from the articular calcified cartilage and subchondral bone. AB - Arthropathy of the distal articular surfaces of the third metacarpal (Mc3) and metatarsal (Mt3) bones in the Thoroughbred racehorse (Tb) is a natural model of repetitive overload arthrosis. We describe a novel pathology that affects the articular calcified cartilage (ACC) and subchondral bone (SCB) and which is associated with hyaline articular cartilage degeneration. Parasagittal slices cut from the palmar quadrant of the distal condyles of the left Mc3/Mt3 of 39 trained Tbs euthanized for welfare reasons were imaged by point projection microradiography, and backscattered electron (BSE) scanning electron microscopy (SEM), light microscopy, and confocal scanning light microscopy. Mechanical properties were studied by nanoindentation. Data on the horses' training and racing career were also collected. Highly mineralised projections were observed extending from cracks in the ACC mineralising front into the hyaline articular cartilage (HAC) up to two-thirds the thickness of the HAC, and were associated with focal HAC surface fibrillation directly overlying their site. Nanoindentation identified this extruded matrix to be stiffer than any other mineralised phase in the specimen by a factor of two. The presence of projections was associated with a higher cartilage Mankin histology score (P<0.02) and increased amounts of gross cartilage loss pathologically on the condyle (P<0.02). Presence of projections was not significantly associated with: total number of racing seasons, age of horse, amount of earnings, number of days in training, total distance galloped in career, or presence of wear lines. PMID- 21623572 TI - Contractile cell forces exerted on rigid substrates. AB - Adhesive cells including fibroblasts produce contractile forces to the underlying substrate for their locomotion. Such forces are not only high enough to deform compliant membranes such as silicone but also bend rigid micro-cantilevers. The cell-induced bending of silicon micro-cantilevers was determined using laser deflection during trypsin treatment. The observed cantilever relaxation corresponds to a contractile cell force of (16+/-7) uN per rat-2 fibroblast. The cantilever bending approach represents a unique method for the determination of contractile cell forces on any kind of rigid substrate in desired (physiological) environment. Hence, the fundamental technique allows building cell-based biosensors or should provide a quantitative parameter for characterising the cyto compatibility of load bearing implant surfaces. PMID- 21623573 TI - A case of giant cell tumor of the hipbone invading the bladder. AB - Giant cell tumor of bone (GCTB) is a very rare neoplasm of the skeleton (about one new case per million population per year). In literature there is a great confusion about GCTB. The majority of authors think that GCTB is a benign locally aggressive tumor, others think that this is a malignant neoplasm and some authors think that GCTB is a reactive condition. This is the first case in literature of GCTB of the hipbone invading the bladder. PMID- 21623574 TI - Home dialysis first: a new paradigm for new ESRD patients. AB - Patients with end-stage renal disease (ESRD) were treated with either in-center hemodialysis (ICH) or one of the modes of home-based dialysis (HBD)-- peritoneal dialysis (PD) or home hemodialysis (HHD). Home-based dialysis modes showed better outcomes than ICH (PD for the first 2-3 years and HHD for the long-term). Home PD has become more attractive with overnight cyclers for PD and the use of home helpers. Home dialysis (PD or HHD) offers a high quality of life and a high degree of independence and is financially attractive. This review will propose a paradigm shift in the initial form of dialysis offered to new patients with ESRD: instead of selecting between in-center dialysis and PD, patients after they are advised of the advantages of dialysis at home (either PD or HHD) should be offered a choice between dialysis at home (PD or HHD) or in hospital. We will review the advantages of home-based dialysis and the arguments for this simple but vital change in the process by which new patients requiring dialysis choose their treatment option. PMID- 21623575 TI - Introductory remarks. PMID- 21623576 TI - Vascular calcification in chronic kidney disease: a changing scenario. AB - Vascular calcification (VC) is one of the most dramatic consequences of chronic kidney disease (CKD). It has been considered a passive process, resulting essentially from mineral metabolism disorders and alterations in calcium and phosphate balance. But during the last decade, it has been elucidated how VC is not only a passive but more properly an active process, in which different factors are deeply involved. The progression of vessel wall mineralization is commonly associated with factors that promote VC, such as age, dialysis vintage and mineral metabolism abnormalities. Furthermore, many substances seem to be dynamically implicated in the regulation of the molecular mechanisms of VC. Between them, the matrix Gla protein and fetuin-A have recently been investigated in CKD. In this review, along with the most promising possible treatments, the new molecular mechanisms involved in the VC process will be elucidated. PMID- 21623577 TI - Vascular calcification in chronic kidney disease: usefulness of a marker of vascular damage. AB - In chronic kidney disease patients, vascular calcification (VC) is a marker of advanced vasculopathy, suggesting both atherosclerotic and medial calcification. Detection and quantification of VC provide physicians with useful prognostic information independent of traditional cardiovascular risk factors. In addition, this knowledge may help guide appropriate therapeutic choices. A number of noninvasive imaging techniques are available to screen for the presence of VC, some with merely qualitative, and others with both qualitative and quantitative, capabilities. We review the strengths and limitations of the most common noninvasive techniques employed to image VC and the prognostic implications of VC. PMID- 21623578 TI - Does any therapy exist for vascular calcifications in uremia? AB - The pathogenesis of vascular calcifications in uremia is not completely understood, but is regarded as multifactorial, involving traditional and nontraditional risk factors. In particular, derangements in divalent ions are considered of outmost importance, but also the role of physiologic inhibitors of calcification is now claimed. The most powerful physiologic inhibitor of calcification is pyrophosphate, but its biochemical instability precludes its clinical use to date. The pharmacologic analogs of pyrophosphate, bisphosphonates, cannot be easily tested for this purpose in renal patients, given their renal clearance. The list of proteins involved in calcification is a growing one, and experimental models point to the potential clinical relevance of matrix Gla protein, fetuin, osteopontin, osteoprotegerin and bone morphogenetic protein-7. Induction of metabolic acidosis, although theoretically useful, is not recommended, while administration of sodium thiosulphate could be beneficial, but its safety awaits confirmation. Actually, the only available therapies for vascular calcifications are those directed toward achievement of the biochemical targets for calcium, phosphate and parathyroid hormone with the hope, but not the certainty, that this will be efficacious. However, to this purpose, selection of the most appropriate strategy in the individual patient seems essential. PMID- 21623579 TI - Update on nephrolithiasis: beyond symptomatic urinary tract obstruction. AB - Nephrolithiasis research and care have been focused on biochemical changes in urinary solute excretion leading to stone formation, but abnormalities in urine chemistry alone do not explain many aspects of the condition of patients with kidney stone disease. Evidence exists of an association with metabolic syndrome, obesity, diabetes and hypertension, and of enhanced risk of chronic kidney disease and metabolic bone disease. Very recently also a higher risk of cardiovascular events and damage has been reported in kidney stone formers when compared with non-stone formers. It is time to view nephrolithiasis as a condition predictive of chronic kidney disease and cardiovascular damage, which deserves full metabolic evaluation together with an early prevention care strategy, mainly consisting of dietary and lifestyle changes, in a multidisciplinary approach. Kidney stone disease should be considered as a systemic disorder with clinical relevance beyond symptomatic urinary tract obstruction. PMID- 21623580 TI - Recent insights into vitamin D and its receptor. AB - The widely differing functions of vitamin D are based both on a wide diffusion of its specific receptor (VDR) and on the ability of many cells, in addition to renal tubular cells, to synthesize calcitriol for autocrine and paracrine functions. In the last few years, many published studies have added new insights into some important points on this topic. Recent data suggest that the control of calcitriol synthesis at tissue levels other than kidneys might differ greatly from the control system working at the renal level. Furthermore, the mechanisms by which the VDR might mediate either the genomic and nongenomic (rapid) vitamin D-mediated effects became much clearer. However, new evidence accumulated suggests that some additional receptor(s), responsive to vitamin D and different from the VDR, could play a role in the rapid response to vitamin D, probably interfering also with the genomic pathway. In this context, there are new possible interpretations of the mechanisms by which different vitamin D metabolites might express variable activities at different levels. In addition, some recent data have challenged the role of the VDR in direct parathyroid hormone (PTH) control, at least in physiological conditions, suggesting that vitamin D-mediated PTH inhibition is mainly secondary to the intestinal effect on calcium absorption. Finally, there is a renewed interest in the field of polymorphic variants in the VDR gene in relation to some clinical conditions, though the mechanisms underlining these associations are far from being clear. The present review briefly addresses all of the above points. PMID- 21623581 TI - New insights into the role of calcium-sensing receptor activation. AB - The discovery of the calcium-sensing receptor (CaSR) prompted the identification of substances that affect its function. Cinacalcet, for example, is a drug that allosterically modifies the receptor so as to increase its sensitivity to circulating calcium (thus the name "calcimimetic") and in this way decreases parathyroid hormone secretion. Clinical use of cinacalcet is already approved for the treatment of primary and secondary hyperparathyroidism, but research is ongoing to identify further potential actions of this drug. The effects and functions of the CaSR have been evaluated in different systems and tissues, beyond parathyroid glands, such arterial walls. A complete understanding of the properties of calcimimetics are of obvious clinical interest, since therapeutic indications may be affected accordingly. PMID- 21623582 TI - CKD-MBD: an endless story. AB - Renal failure is a growing problem that involves a large part of the population and has a great social impact, with often incapacitating complications, mainly related to mineral bone disorders (MBD) and cardiovascular diseases. Analysis of the recent scientific literature confirms that a large number of chronic kidney disease (CKD) patients develop an early derangement of the parameters of Ca-P metabolism in which phosphate homeostasis and a reduced endogenous synthesis of calcitriol play a critical role. Recent findings from several large observational studies have also suggested that the benefits of vitamin D receptor activators may extend beyond the traditional parathyroid hormone-lowering effect, and could result in direct cardiovascular and metabolic benefits. Treatment of secondary hyperparathyroidism has become even more complex with the arrival of the calcium sensing receptor agonist cinacalcet hydrochloride and with the uncovering of novel mechanisms responsible for secondary hyperparathyroidism. The aim of this review is the analysis of some of the recent contributions in the field of CKD MBD, to update the understanding of the pathogenetic mechanisms and possibly the most appropriate therapeutic approach in this field. PMID- 21623583 TI - TRAIN: Training through Research Application Italian iNitiative. AB - Training through Research Application Italian iNitiative (TRAIN) is a mobility program financed under the EU action called "Cofinancing of regional, national and international programs" (COFUND) of the European Commission Seventh Framework Program (FP7) - People, and has been designed to encourage the promotion and development of international programs of research through mobility at various stages of research careers. The aim of TRAIN is to improve translational skills in the field of cancer by promoting a three-year international mobility program assigning a total of 51 fellowships subdivided into incoming, outgoing and reintegration fellowships.?The TRAIN proposal has been submitted in February 2009 to the European Commission in reply to the 2008 FP7-PEOPLE-COFUND call and has been successfully evaluated. TRAIN is addressed to postdoctoral scientists or scientists who have at least four years' full-time equivalent research experience and who wish to improve their careers spending one year abroad. The mobility program is open also to non-Italian experienced scientists wishing to spend one year in an Italian research center or private company. Part of the scheme is targeted to experienced Italian scientists who have completed at least three years of research in a foreign country and are interested in returning to Italy.?TRAIN is part of an overall Italian strategy outlined by the International Program of the Italian Cancer Network "Alleanza Contro il Cancro" to promote Italian participation in the building of the European Area for translational cancer research and to enhance the interaction between academy and industry. PMID- 21623584 TI - One size fits all in prostate cancer: a story tale whose time has come and gone. AB - The touchstone to evaluate accurately the aggressiveness and invasiveness of prostate cancer is something of a holy grail in the facet of urologic oncology. Gene expression and sequencing studies have improved our interpretations of the genetic determinants of the disease but are unsuccessful in the establishment of any unified classification to improve the molecular stratification. These questions addressing failure in rational drug design are difficult to answer in the multifaceted and heterogeneous pathogenesis of prostate cancer. In this review, we have developed a roadmap of the "recalcitrant prostate cancer proteome" to recognize the aspects of prostate cancer that may be helpful in effectively translating these findings to the clinic. PMID- 21623585 TI - Perfusion circuit concepts for hollow-fiber bioreactors used as in vitro cell production systems or ex vivo bioartificial organs. AB - For the development and implementation of primary human cell- and stem cell-based applications in regenerative medicine, large amounts of cells with well-defined characteristics are needed. Such cell quantities can be obtained with the use of hollow fiber-based bioreactors. While the use of such bioreactors generally requires a perfusion circuit, the configuration and complexity of such circuits is still in debate. We evaluated various circuit configurations to investigate potential perfusate volume shifts in the arterial and venous sides of the perfusion circuit, as well as in the feed and waste lines. Volume shifts with changes in flow conditions were measured with graduated bubble traps in the circuit, and perfusion pressures were measured at three points in the circuits. The results of this study demonstrate that the bioreactor perfusion circuit configuration has an effect on system pressures and volume shifts in the circuit. During operation, spikes in post-bioreactor pressures caused detrimental, potentially dangerous volume shifts in the feed and waste lines for configurations that lacked feed pumps and/or waste line check valves. Our results indicate that a more complex tubing circuit adds to safety of operation and avoids technical challenges associated with the use of large-scale hollow fiber bioreactors (e.g., for extracorporeal liver support or erythrocyte production from hematopoietic stem cells), including volume shifts and the need for a large reservoir. Finally, to ensure safe use of bioreactors, measuring pre-, intra-, and post-bioreactor pressures, and pump operation control is also advisable, which suggests the use of specifically developed bioreactor perfusion devices. PMID- 21623586 TI - PUCA pump and IABP comparison: analysis of hemodynamic and energetic effects using a digital computer model of the circulation. AB - The pulsatile catheter pump (PUCA pump) is a left ventricular assist device that provides additional flow to the left ventricle. It is usually run in order to ensure a counterpulsation effect, as in the case of the intra-aortic balloon pump (IABP). Because of this similarity, a comparison between the PUCA pump and the IABP was conducted from both the hemodynamic and energetic points of view. Numerical models of the two devices were created and connected to the CARDIOSIM cardiovascular simulator. The PUCA and IABP models were then verified using in vivo experimental data and literature data, respectively. Numerical experiments were conducted for different values of left ventricular end systolic elastance (Els) and systemic arterial compliance (Csa). The energetic comparison was conducted taking into account the diastolic pressure time index and the endocardial viability ratio. Hemodynamic results expressed as cardiac output (CO) and mean coronary blood flow (CBF) show that both the IABP and the PUCA pump efficacy decrease with higher values of Els and Csa. The IABP especially shows higher sensitivity to these parameters, to the extent that in some cases CO actually drops and CBF does not increase. On the other hand, for lower values of Csa, IABP performance improves so much that the PUCA pump flow needs to be increased in order to ensure a hemodynamic effect comparable to that of the IABP. Energetic results show a trend similar to the hemodynamic ones. The study will be continued by investigating other energetic variables and the autonomic response of the cardiovascular system. PMID- 21623587 TI - HSP induction in mesothelial cells by peritoneal dialysis fluid depends on biocompatibility test system. AB - BACKGROUND: We have previously shown that exposure of mesothelial cells (MC) to peritoneal dialysis fluids (PDF) not only caused toxic injury, but also induced cytoprotective heat shock proteins (HSP). This study was performed in order to compare HSP expression in MC upon PDF exposure in three currently used biocompatibility test systems. METHODS: Omentum-derived human peritoneal MC underwent 3 modalities of exposure to heat- or filter-sterilized PDF: (A) pure PDF for 60 minutes followed by a recovery-period in pure culture medium for 24 hours; (B) 1:1 mixture of PDF and culture medium for 24 hours or (C) pure PDF for 60 minutes followed by a recovery-period in a 1:1 mixture of PDF and culture medium for 24 hours. Biocompatibility was assessed by LDH-release into the supernatant and HSP-72 expression in MC lysates. RESULTS: Short-term exposure of MC to pure PDF (Modality A) resulted in concordant LDH release and upregulation of HSP-72, regardless of heat or filter sterilization. In contrast, both test systems that exposed MC to heat-sterilized PDF during the recovery period (Modalities B and C) resulted in severe cellular lethality but low HSP-72 expression. CONCLUSIONS: This study clearly shows that HSP expression in MC upon PDF exposure depends on the biocompatibility test system. The presence of heat sterilized PDF during recovery resulted in significant downregulation of Hsp-72 despite severe cell injury. Therefore, Hsp-72 expression reflects adequate cellular stress responses rather than PDF cytotoxicity. PMID- 21623588 TI - Nonorganic visual loss and associated psychopathology in children. AB - PURPOSE: To report the frequency of nonorganic visual loss (NOVL) and associated psychopathology in children. METHODS: A total of 973 children were examined in our ophthalmology practice between 2006 and 2009. Basic ophthalmologic exploration (visual acuity, stereopsis, cycloplegic refraction, ocular motility, pupil dynamics, biomicroscopy, indirect ophthalmoscopy) and specific tests for NOVL diagnosis were performed (confusion with lenses test, mirrors test, Roth test, Bravais test). We also investigated the psychosocial situation and associated psychiatric problems. RESULTS: Thirty children were diagnosed with NOVL. The mean age of the children was 8.93 years (+/- 2.61); 70% were girls. September was the commonest month of presentation (26.7%) and unilateral (3.3%) or bilateral (80%) visual loss was the most frequent symptom (83.3% in total). In 20% of cases we detected psychosocial anomaly and 40% were seeking to wear glasses. CONCLUSIONS: Malingering in children is very frequent. We can make the diagnosis with simple tests. It is not necessary to perform imaging and electrophysiologic testing, thus avoiding unnecessary examinations as well as absenteeism from work for parents and health care costs. PMID- 21623589 TI - SD OCT features of dry arcuate longstanding retinal folds. PMID- 21623590 TI - Visual acuity, contrast sensitivity, subjective quality of vision, and quality of life with 4 different multifocal IOLs. AB - PURPOSE: This prospective, randomized, double-masked, clinical trial was designed to evaluate visual acuity, contrast sensitivity, subjective quality of vision, and quality of life in 47 patients symmetrically, and randomly, implanted with 4 different IOL designs (SN6AD1, SN60D3, ReZoom NXG1, and Tecnis ZMA00), 3 months after cataract intervention. METHODS: Binocular corrected and uncorrected distance visual acuity, binocular distance corrected (BCNVA) and uncorrected (UCNVA) near visual acuity, binocular distance corrected (BCIVA) and uncorrected (UCIVA) intermediate visual acuity, photopic, mesopic, and mesopic with glare contrast sensitivity, quality of vision, and quality of life were evaluated. RESULTS: Statistically and clinically significant differences were found in BCNVA and UCNVA at 33 cm between high and low add power IOLs, while diffractive SN6AD1 lenses achieved better UCNVA at 40 cm than refractive ReZoom IOLs. Asphericity and low add power were found to improve BCIVA. Contrast sensitivity was similarly compromised in all IOL models, although diffractive optics and aspheric profiles performed better in mesopic conditions. All IOL types received similar overall satisfaction and quality of life scores. Whereas ReZoom patients depended on their spectacles for near tasks, intermediate vision was spectacle independent. Photic phenomena were present in all IOLs, albeit more frequent in ReZoom IOLs. CONCLUSIONS: The present results, which reflect IOL characteristics in optics, profile, and add power, may contribute to help surgeons decide on the type of IOL most suitable for each patient, especially those with high visual demands at near and intermediate distances. PMID- 21623591 TI - Cataract as a phenotypic marker for a mutation in WFS1, the Wolfram syndrome gene. AB - PURPOSE: Wolfram syndrome (WS) or diabetes insipidus, diabetes mellitus, optic atrophy, and deafness (DIDMOAD) (OMIM 222300) is an inherited neurodegenerative disease characterized by diabetes mellitus and optic atrophy as the 2 major criteria, followed later in life by deafness, diabetes insipidus, and various signs of neurologic impairment. The presence of a cataract has been variably mentioned in WS. METHOD: Two members of a family had thorough ophthalmic examination and their DNA was screened for mutations in mitochondrial DNA, WFS1, OPA1, and OPA3 genes. RESULTS: We report a patient who first had surgery for bilateral cataract at age 5 and who subsequently presented typical signs of WS, i.e., diabetes mellitus, optic atrophy with reduced visual acuity at 20/400 on both eyes at age 22, and mild deafness. The patient was found to be a compound heterozygote for 2 truncating mutations in WFS1, the major WS gene. She carried the previously reported c.1231_1233 delCT and a novel c.2431_2465dup35 mutation. She also was heterozygote for a novel OPA1 sequence variant, c.929A>G in exon 9, whose pathogenicity remains uncertain. The patient's mother was a heterozygous carrier of the c.2431_2465dup35 mutation. She did not have diabetes mellitus or optic atrophy but had bilateral polar cataract. She did not carry the OPA1 sequence variant. CONCLUSIONS: Cataract could be a marker for the WFS1 heterozygosity in this family, namely the c.2431_2465dup35 mutation. PMID- 21623593 TI - Precision, morphology, and histology of corneal flap cuts using a 200-kHz femtosecond laser. AB - PURPOSE: Laser-assisted in situ keratomileusis (LASIK) requires precise corneal flap cutting. Especially the creation of thin flaps has recently gained importance for thin-flap LASIK. Currently, there is a trend towards faster femtosecond lasers that can produce flaps in a short period of time. We analyzed flaps created with a 200-kHz femtosecond laser concerning their cut precision, morphology, and histology. METHODS: Femtosecond laser flap cutting was performed on 36 porcine cadaver eyes using the prototype 200-kHz femtosecond laser UltraFlap (WaveLight GmbH, Erlangen, Germany). The eyes were assigned to 3 thickness groups, with a cut depth of 100 um, 130 um, or 180 um, respectively. Additionally, flap diameters were varied, ranging from 8.0 mm to 9.5 mm. Flap thicknesses were determined with a micrometer gauge. Flap diameters were measured with a sliding caliper. Furthermore, flaps were created for histologic examination. RESULTS: There were no complications during flap creation. The mean flap thickness and standard deviation was (in micrometers) 96.33 +/- 7.45 (intended thickness: 100), 134.67 +/- 4.96 (intended thickness: 130), and 174.59 +/- 9.35 (intended thickness: 180), respectively. The flap diameter revealed a mean (in mm) of 8.03 +/- 0.15 (intended diameter: 8.0), 8.56 +/- 0.10 (intended diameter: 8.5), 9.09 +/- 0.10 (intended diameter: 9.0), and 9.54 +/- 0.15 (intended diameter: 9.5), respectively. Histologic examination showed very little to almost no changes in the structure of the corneal stroma. CONCLUSIONS: Flap creation could be performed easily without any complications. The morphology and accuracy of the cuts were very reliable and precise. Histology showed a smooth cut. PMID- 21623594 TI - Somatotopic arrangement and location of the corticospinal tract in the brainstem of the human brain. AB - The corticospinal tract (CST) is the most important motor pathway in the human brain. Detailed knowledge of CST somatotopy is important in terms of rehabilitative management and invasive procedures for patients with brain injuries. In this study, I conducted a review of nine previous studies of the somatotopical location and arrangement at the brainstem in the human brain. The results of this review indicated that the hand and leg somatotopies of the CST are arranged medio-laterally in the mid to lateral portion of the cerebral peduncle, ventromedial-dorsolaterally in the pontine basis, and medio-laterally in the medullary pyramid. However, few diffusion tensor imaging (DTI) studies have been conducted on this topic, and only nine have been reported: midbrain (2 studies), pons (4 studies), and medulla (1 study). Therefore, further DTI studies should be conducted in order to expand the literature on this topic. In particular, research on midbrain and medulla should be encouraged. PMID- 21623595 TI - Current status of the Korean venous thromboembolism registry. AB - The Korean venous thromboembolism (VTE) registry, which was initiated by the Working Parties of Korean Society on Thrombosis and Hemostasis, and the Korean Society of Hematology, is a web-based multicenter registry (http://kdvt.chamc.co.kr) for recruiting consecutive VTE patients. The aim of the registry is to prospectively collect data on the epidemiology and clinical outcomes of VTE from a large, unselected cohort of patients, and to provide data on the true incidence and management of VTE in the real-world. By the end of 2007, the starting year of the registry, 840 patients were registered. By the end of 2008, 1,121 were registered, with 1,289 by the end of 2009, and 1,463 by April 2010 from 11 hospitals. The first report on the epidemiologic characteristics of 596 consecutive VTE patients was released in October 2007. PMID- 21623596 TI - Gender-based differences in the management and prognosis of acute coronary syndrome in Korea. AB - PURPOSE: Gender-based differences exist in the characteristics, management, and prognosis of acute coronary syndrome (ACS). However, their impact on prognosis remains unclear. We aimed to identify factors causing these differences in Koreans. MATERIALS AND METHODS: We examined 6,636 ACS patients (66.2% males) visiting 72 Korean hospitals between April-2007 and December-2008. Gender-based differences in clinical demographics, therapy, and outcomes were analyzed over 6 months. RESULTS: Women were older than men [mean (standard deviation, SD) age, 67.6 (9.8) vs. 60.6 (11.2) years; p<0.001]; had higher rates of hypertension, diabetes mellitus, and lack of exercise (p<0.001 for all); and lower rates of obesity, familial history of cardiovascular disease (CVD), and smoking (p<0.05 for all). Atypical symptoms were more common in women (20.5% vs. 15.1% in men, p<0.001), whereas myocardial infarction with ST-segment elevation was less common (17.1% vs. 27.8%, p<0.001). Mean (SD) time lapse from symptom onset to arrival at hospital was longer in women [11.44 (18.19) vs. 8.26 (14.89) hours in men, p<0.001], as was the duration of hospitalization [7.58 (7.61) vs. 7.04 (7.72) days, p=0.007]. Fewer women underwent revascularization procedures, including thrombolytic therapy, balloon angioplasty, stent implantation, and coronary artery bypass grafting (79.4% vs. 83.3% men, p<0.001). No significant differences were observed in CVD-related death, recurrent ACS, stroke, refractory angina, or rehospitalization for angina. CONCLUSION: Female ACS patients were older than male subjects and had more atypical presentation. They arrived at the hospital later than men and had longer hospital stays, but less often required revascularization therapy. However, no gender-based differences were noted in ACS related mortality and morbidity. PMID- 21623597 TI - Effects of posture and acute sleep deprivation on heart rate variability. AB - PURPOSE: In our previous study to investigate autonomic nervous system (ANS) activity due to radio frequency (RF) radiation using heart rate variability (HRV), drowsiness was observed in approximately half of all subjects. Therefore, the usage of HRV with unwanted drowsiness could falsely indicate the effects of RF radiation by mobile phones on the ANS. The objective of this study was to determine which posture is appropriate for accurate HRV analysis for provocation study. MATERIALS AND METHODS: A total of 52 healthy subjects (25 males and 27 females) participated in this experiment. We measured the number of times a subject showed drowsiness or sleep deprivation due to awakening, and analyzed HRV six times over 30 minutes in sitting and recumbent postures, using power spectrum. RESULTS: We employed the ratio of low frequency power to high frequency power (LFP/HFP) to analyze the changes in the ANS. The number of sleep deprivation occurrences in the sitting posture was significantly less than that in the recumbent posture (p<0.01), resulting in smaller increase of LFP/HFP. Although LFP/HFP of the two postures varied with time without any provocation, it was more stable in sitting than in recumbent postures. CONCLUSION: A sitting posture is preferable to a recumbent posture for analyzing HRV, because of decreased drowsiness and sleep deprivation, thereby decreasing variation of LFP/HFP during experiment. Considering the drowsiness, it is also recommended that any experiment should be completed within 15 minutes, if possible. PMID- 21623598 TI - Using multidetector-row CT for the diagnosis of afferent loop syndrome following gastroenterostomy reconstruction. AB - PURPOSE: To assess the clinical manifestations and multidetector-row computed tomography (MDCT) findings of afferent loop syndrome (ALS) and to determine the role of MDCT on treatment decisions. MATERIALS AND METHODS: From January 2004 to December 2008, 1,100 patients had undergone gastroenterostomy reconstruction in our institution. Of these, 22 (2%) patients were diagnosed as ALS after surgery that included Roux-en-Y gastroenterotomy (n=9), Billroth-II gastrojejunostomy (n=7), and Whipple's operation (n=6). Clinical manifestations and MDCT features of these patients were recorded and statistically analyzed. The presumed etiologies of obstruction shown on the MDCT were correlated with clinical information and confirmed by surgery or endoscopic biopsy. RESULTS: The most common clinical symptom was acute abdominal pain, presenting in 18 patients (82%). We found that a fluid-filled C-shaped afferent loop in combination with valvulae conniventes projecting into the lumen was the most common MDCT features of ALS. Malignant causes of ALS, such as local recurrence and carcinomatosis, are the most common etiologies of obstruction. These etiologies and associated complications can be predicted 100% by MDCT. CONCLUSION: Our results suggest that MDCT is a reliable modality for assessing the etiologies of ALS and guiding treatment decisions. PMID- 21623599 TI - Apoptosis of T lymphocytes isolated from peripheral blood of patients with acute exacerbation of chronic obstructive pulmonary disease. AB - PURPOSE: Chronic obstructive pulmonary disease (COPD) is characterized by chronic inflammation of the airways and progressive destruction of lung parenchyma. Apoptosis is critical for the maintenance of normal tissue homeostasis and is in equilibrium with proliferation and differentiation. This study was undertaken to investigate relationship between apoptosis of peripheral blood lymphocytes during exacerbation of COPD and inflammatory response that characterizes this condition. MATERIALS AND METHODS: Seventeen patients with COPD exacerbation, 21 stable COPD, and 12 control subjects were included. T lymphocytes were isolated from peripheral blood using MACS. Apoptosis of T lymphocytes was assessed with FACS using annexin V and 7-aminoactinomycin. Serum levels of interleukin (IL)-6, IL-8 and tumor necrosis factor (TNF)-alpha were determined by an immunoassay technique. RESULTS: There was significantly increased percentage of apoptotic lymphocytes, CD 4+, and CD 8+ T cells in the peripheral blood of patients with exacerbation of COPD compared with stable COPD. Serum levels of IL-6, IL-8, and TNF-alpha were significantly increased in patients with exacerbation of COPD compared with stable COPD. Only TNF-alpha presented a positive correlation with apoptotic lymphocytes in patients with exacerbation of COPD. CONCLUSION: Increased apoptotic lymphocytes may be associated with upregulation of TNF-alpha in the peripheral blood of patients with acute exacerbation of COPD. PMID- 21623600 TI - Anti-proteinuric effect of sulodexide in immunoglobulin a nephropathy. AB - PURPOSE: We conducted a multi-center randomized double-blind study to determine the effects of 6-month therapy with sulodexide on urinary protein excretion in patients with idiopathic Immunoglobulin A (IgA) nephropathy. MATERIALS AND METHODS: A total of seventy-seven patients participated in the study. They were randomly allocated to one of three groups: sulodexide 75 mg or 150 mg daily or the placebo for 6 months. The primary end point was the achievement, at 6 months, of at least 50% reduction in urine protein/creatinine ratio (UPCR) from the baseline value. RESULTS: At 6 months, the primary end point was achieved by 12.5% of the patients assigned to the placebo, 4.0% of the patients assigned to sulodexide 75 mg daily and 21.4% of those assigned to 150 mg (p=0.308). Treatment with sulodexide 150 mg daily for 6 months significantly reduced log UPCR from 6.38+/-0.77 at baseline to 5.98+/-0.94 at 6 months (p=0.045), while treatment with sulodexide 75 mg daily and placebo did not. CONCLUSION: A 6-month treatment with sulodexide did not achieve 50% reduction of urinary protein excretion in IgA nephropathy patients, but showed a tendency to increase the time-dependent anti proteinuric effect. Therefore, long-term clinical trials on a larger scale are warranted to elucidate the hypothesis that sulodexide affords renal protection in IgA nephropathy patients. PMID- 21623601 TI - A more appropriate cardiac troponin T level that can predict outcomes in end stage renal disease patients with acute coronary syndrome. AB - PURPOSE: Cardiac troponin T (cTnT), a useful marker for diagnosing acute myocardial infarction (AMI) in the general population, is significantly higher than the usual cut-off value in many end-stage renal disease (ESRD) patients without clinically apparent evidence of AMI. The aim of this study was to evaluate the clinical usefulness of cTnT in ESRD patients with acute coronary syndrome (ACS). MATERIALS AND METHODS: Two hundred eighty-four ESRD patients with ACS were enrolled between March 2002 and February 2008. These patients were followed until death or June 2009. Medical records were reviewed retrospectively. The cut-off value of cTnT for AMI was evaluated using a receiver operating characteristic (ROC) curve. We calculated Kaplan-Meier survival curves, and potential outcome predictors were determined by Cox proportional hazard analysis. RESULTS: AMIs were diagnosed in 40 patients (14.1%). The area under the curve was 0.98 in the ROC curve (p<0.001; 95% CI, 0.95-1.00). The summation of sensitivity and specificity was highest at the initial cTnT value of 0.35 ng/mL (sensitivity, 0.95; specificity, 0.97). Survival analysis showed a statistically significant difference in all-cause and cardiovascular mortalities for the group with an initial cTnT >=0.35 ng/mL compared to the other groups. Initial serum cTnT concentration was an independent predictor for mortality. CONCLUSION: Because ESRD patients with an initial cTnT concentration >=0.35 ng/mL have a poor prognosis, it is suggested that urgent diagnosis and treatment be indicated in dialysis patients with ACS when the initial cTnT levels are >=0.35 ng/mL. PMID- 21623602 TI - Impact of acute kidney injury on clinical outcomes after ST elevation acute myocardial infarction. AB - PURPOSE: This study aimed to compare the incidence and clinical significance of transient versus persistent acute kidney injury (AKI) on acute ST elevation myocardial infarction (STEMI). MATERIALS AND METHODS: The study was a retrospective cohort of 855 patients with STEMI. AKI was defined as an increase of >=0.3 mg/dL in creatinine level at any point during hospital stay. The study population was classified into 5 groups: 1) patients without AKI; 2) patients with mild AKI that was resolved by discharge (creatinine change less than 0.5mg/dL compared with admission creatinine during hospital stay, transient mild AKI); 3) patients with mild AKI that did not resolve by discharge (persistent mild AKI); 4) patients with moderate/severe AKI that was resolved by discharge (creatinine change more than 0.5 mg/dL compared with admission creatinine, transient moderate/severe AKI); 5) patients with moderate/ severe AKI that did not resolve by discharge (persistent moderate/severe AKI). We investigated 1-year all-cause mortality after hospital discharge for the primary outcome of the study. The relation between AKI and 1-year mortality after STEMI was analyzed. RESULTS: AKI occurred in 74 (8.7%) patients during hospital stay. Adjusted hazard ratio for mortality was 3.139 (95% CI 0.764 to 12.897, p=0.113) in patients with transient, mild AKI, and 8.885 (95% CI 2.710 to 29.128, p<0.001) in patients with transient, moderate/severe AKI compared to patients without AKI. Persistent moderate/severe AKI was also independent predictor of 1 year mortality (hazard ratio, 5.885; 95% CI 1.079 to 32.101, p=0.041). CONCLUSION: Transient and persistent moderate/severe AKI during acute myocardial infarction is strongly related to 1-year all cause mortality after STEMI. PMID- 21623603 TI - Toll-like receptor 4 signaling is involved in IgA-stimulated mesangial cell activation. AB - PURPOSE: Deposition of polymeric IgA1 in the kidney mesangium is the hallmark of IgA nephropathy, but the molecular mechanisms of IgA-mediated mesangial responses and inflammatory injuries remain poorly understood. We hypothesize that Toll-like receptor 4 (TLR4) is involved in IgA-induced mesangial cell activation. MATERIALS AND METHODS: Mouse mesangial cells were stimulated with lipopolysaccharide (LPS) (1 MUg/mL), IgA (20 MUg/mL), or both, and TLR4 expression was measured by real time RT-PCR and Western blot. Intracellular responses to LPS or IgA were assessed by Western blot for ERK1/2, JNK, p38 MAP kinases (MAPKs), Ikappa-Balpha degradation and fibronectin secretion. MCP-1 secretion was assessed by ELISA. Small interfering RNA (siRNA) of TLR4 was used to confirm that the effects were caused by TLR4 activity. RESULTS: LPS- or IgA-treatment upregulated the levels of TLR4 mRNA and protein in cultured MMC at 24 h. LPS and IgA induced rapid phosphorylation of MAPKs, but degradation of Ikappa-Balpha was observed only in LPS-treated MMC. LPS, but not IgA, induced increased secretion of MCP-1 and fibronectin at 24 h or 48 h. Combined LPS and IgA treatment did not cause additional increases in TLR4 mRNA and protein levels or Ikappa-Balpha degradation, and MCP-1 and fibronectin secretions were less than with LPS alone. LPS- or IgA-induced TLR4 protein levels and MAPK activation were inhibited by transfection with TLR4 siRNA. CONCLUSION: These results indicate that the activation of MAPKs and MCP-1 secretion are mediated by TLR4, at least in part, in IgA-treated mesangial cells. TLR4 is involved in mesangial cell injury by induction of pro-inflammatory cytokines in IgA nephropathy. PMID- 21623604 TI - Teicoplanin dosing strategy for treatment of Staphylococcus aureus in Korean patients with neutropenic fever. AB - PURPOSE: The present study was conducted to determine and compare the target attainment rate (TAR) between microorganism-nonspecific (C(trough)) and microorganism- specific (AUC24/MIC) targets over two weeks of teicoplanin administration according to several dose regimens for the treatment of Staphylococcus aureus in Korean patients with neutropenic fever. MATERIALS AND METHODS: One thousand virtual concentrations were obtained for each dose using the population pharmacokinetic parameters of teicoplanin adopted from a published study. Simulation of 1,000 virtual MICs was performed using the MICs of 78 clinical isolates of S. aureus collected from a hospital in Korea. Thereafter, these simulated MICs were randomly allocated to 1,000 virtual patients in whom the TARs for AUC24/MIC>125 [or 345] and C(trough)>10 [or 20] mg/L were determined. The relationship of the maintenance dose with the steady-state TAR was predicted with respect to the AUC24/MIC>125 [or 345] using logistic analysis. RESULTS: The standard dose regimen of teicoplanin showed TARs of about 70% [or 33%] and 70% [or 20%] at steady-state in cases with AUC24/MIC>125 [or 345] and C(trough)>10 [or 20] mg/L, respectively. CONCLUSION: The current standard dose regimen was predicted to be insufficient to adequately treat S. aureus in Korean patients with neutropenic fever. To assure at least an 80% TAR in this population, dose adjustment of teicoplanin should be considered. PMID- 21623605 TI - Lack of effect of dexamethasone on growth of Orientia tsutsugamushi Gilliam in mouse L929 cells. AB - PURPOSE: Previous studies and our own clinical experience suggest that concurrent corticosteroid treatment for severe rickettsial disease with multiorgan failure may improve the clinical course or reduce mortality. However, the use of corticosteroids as adjunctive treatment for rickettsial diseases is controversial. We attempted to determine the influences of corticosteroid on the growth of Orientia tsutsugamushi in vitro to justify and evaluate the clinical applicability of corticosteroid in rickettsial disease. MATERIALS AND METHODS: L929 cells were infected with Orientia tsutsugamushi Gilliam. Dexamethasone was added to the cells at final concentrations of 101 and 107 pg/mL. Cultures were incubated at 35 degrees C and processed for flow cytometry on the 6th day after addition of dexamethasone. RESULTS: Observation on the 6th day after treatment with dexamethasone in infected cultures revealed that there was no difference in fluorescence intensity among the treatment wells. Treatment of the cells with dexamethasone at concentrations of 101 and 107 pg/mL showed no influence on the growth of Orientia tsutsugamushi. CONCLUSION: Our results to show that isolated corticosteroid does not enhance the replication of Orientia tsutsugamushi in vitro. Concurrent use of anti-inflammatory or immunosuppressive doses of corticosteroids in conjunction with antibiotics may not have detrimental effects on the course of scrub typhus. PMID- 21623606 TI - Clinical review of endogenous endophthalmitis in Korea: a 14-year review of culture positive cases of two large hospitals. AB - PURPOSE: To identify the clinical features and outcomes of endogenous endophthalmitis in Korea. MATERIALS AND METHODS: We reviewed 18 patients with endogenous endophthalmitis at 2 Korean hospitals, treated over a 14 year period between January 1993 and December 2006. RESULTS: The comorbidities observed in these cases were diabetes mellitus and liver cirrhosis. The most common pathogens, which were found in 7 patients each (38.9%), were Klebsiella pneumonia and Pseudomonas aeruginosa. All patients were treated with systemic antibiotics and fortified topical antibiotics. A surgical approach including vitrectomy was performed in 9 cases (50.0%). The prognosis was generally poor, and visual acuity improved slightly in 6 patients (33.3%). CONCLUSION: In this study, diabetes mellitus and Klebsiella pneumonia showed a close relationship with endogenous endophthalmitis, respectively. Endogenous endophthalmitis is a serious risk to sight and careful attention to establishing the diagnosis and management may decrease the ocular morbidity. PMID- 21623607 TI - Inflammatory and tumor stimulating responses after laparoscopic sigmoidectomy. AB - PURPOSE: Laparoscopic colectomy has clinical benefits such as short hospital stay, less postoperative pain, and early return of bowel function. However, objective evidence of its immunologic and oncologic benefits is scarce. We compared functional recovery after open versus laparoscopic sigmoidectomy and investigated the effect of open versus laparoscopic surgery on acute inflammation as well as tumor stimulation. MATERIALS AND METHODS: A total of 57 patients who were diagnosed with sigmoid colon cancer were randomized for elective conventional or laparoscopically assisted sigmoidectomy. Serum samples were obtained preoperatively and on postoperative day 1. C-reactive protein (CRP) and interleukin-6 (IL-6) were measured as inflammation markers, and vascular endothelial growth factor (VEGF) and insulin-like growth factor binding protein-3 (IGFBP-3) were used as tumor stimulation factors. Clinical parameters and serum markers were compared. RESULTS: Postoperative hospital stay (p=0.031), the first day of gas out (p=0.016), and the first day of soft diet (p<0.001) were significantly shorter for the laparoscopic surgery group than the open surgery group. The levels of CRP, IL-6, and VEGF rose significantly, and the concentration of IGFBP-3 fell significantly after both open and laparoscopic surgery. However, there were no significant differences in the preoperative and postoperative levels of CRP, IL-6, VEGF, and IGFBP-3 between the two groups. CONCLUSION: Our data suggest that both open and laparoscopic surgeries are accompanied by significant changes in IL-6, CRP, IGFBP-3, and VEGF levels. Acute inflammation markers and tumor stimulating factors may not reflect clinical benefits of laparoscopic surgery. PMID- 21623608 TI - Endoscopic thyroidectomy via an axillo-breast approach without gas insufflation for benign thyroid nodules and micropapillary carcinomas: preliminary results. AB - PURPOSE: To examine the feasibility of endoscopic thyroidectomy (ET) via an axillo- breast approach without gas insufflation for large thyroid tumors and micropapillary carcinomas. MATERIALS AND METHODS: The patients in the benign group were separated into groups 1 (n=95, <4 cm in tumor diameter) and 2 (n=37, >=4 cm in tumor diameter). Also, 57 patients in the micropapillary carcinoma group underwent an endoscopic hemithyroidectomy (HT) (group 3) and were compared with 60 patients who received conventional open HT (group 4). Postoperative functional outcome, local complications, surgical outcomes, and pathological outcomes were compared between the groups. RESULTS: In the benign group, there was no significant difference in mean operating time, hospital stay, or overall perioperative complications between the two groups. In the micropapillary carcinoma group, mean operating time and hospital stay in group 3 were significantly longer than in group 4 (p=0.015 and p<=0.001). The overall perioperative complications did not differ significantly between the groups. The postoperative cosmetic result was better in groups 1-3 (endo group) than in group 4 (open group). CONCLUSION: ET via a gasless axillo-breast approach seems to be a safe procedure even for benign thyroid lesions >=4 cm and micropapillary carcinomas. Although it has the advantage of better cosmetic results over open thyroidectomy, there is room for improvement in terms of lessening its invasiveness and shortening the operative time. PMID- 21623609 TI - Total hip arthroplasty using S-ROM prosthesis for dysplastic hip. AB - PURPOSE: The purpose of this study was to evaluate the clinical and radiological results of total hip arthroplasty using a proximal modular femoral stem in patients who had secondary coxarthrosis associated with a dysplastic hip. MATERIALS AND METHODS: Forty-two patients (45 hips) with secondary coxarthrosis were evaluated after undergoing primary total hip arthroplasty using an S-ROM proximal modular femoral stem. The average follow-up was 80 months (range: 60 to 96 months). Clinical and radiological assessments were performed based on the Harris hip score and the radiological changes around the prosthesis. RESULTS: The average Harris hip score improved from 52.2 points to 88.5 points. All femoral stems showed stable fixation; there were 37 cases by bony ingrowth and 8 cases by stable fibrous ingrowth. Neither osteolysis nor progressive radiolucent lines around the femoral stem were found at the last follow-up. Forty-one hips (91.9%) revealed excellent or good clinical results at the most recent follow-up. CONCLUSION: For advanced secondary coxarthrosis, total hip arthroplasty with the use of the proximal modular femoral stem yielded good mid-term results with respect to the clinical and radiological criteria. PMID- 21623610 TI - Causes of shoulder pain in women with breast cancer-related lymphedema: a pilot study. AB - PURPOSE: To inform on shoulder pathology and to identify the disabilities and level of quality of life (QOL) associated with shoulder pain in patients with breast cancer- related lymphedema (BCRL). MATERIALS AND METHODS: Using patient history, physical examination, and ultrasound (US), we classified patients with BCRL into the following three groups: no pain with normal ultrasound (US), pain with normal US, and pain with abnormal US. We evaluated shoulder pathology using US, pain intensity using a visual analogue scale (VAS), and functional disability using the Korean version of the Disabilities of Arm, Shoulder, and Hand (DASH) questionnaire. For assessment of QOL, we used the Korean version of the brief form of the World Health Organization Quality of Life Assessment Instrument (WHOQOL- BREF). RESULTS: 28.9% of patients had no pain and normal US, 31.6% had pain with normal US, and 39.5% had pain with abnormal US. The US findings for those with pain and abnormal US revealed the following: 53.3% had a supraspinatus tear, 13.3% had biceps tenosynovitis, 13.3% had acromioclavicular arthritis, 13.3% had subdeltoid bursitis, and 53.3% had adhesive capsulitis. Patients with shoulder pain and abnormal US findings had significantly higher mean DASH and pain scores. Pain scores were positively correlated with DASH scores and negatively correlated with QOL. CONCLUSION: We found that BCRL with shoulder pain and evidence of shoulder pathology on US was associated with reduced QOL and increased disability. Proper diagnosis and treatment of shoulder pain are necessary to improve QOL and decrease disability in patients with BCRL. PMID- 21623611 TI - Effect of charcoal filter on the emergence from sevoflurane anesthesia in a semi closed rebreathing circuit. AB - PURPOSE: A charcoal filter attached within the anesthetic circuit has been shown to efficiently adsorb halothane or isoflurane, thus hastening anesthetic recovery in low or minimal flow system. This study was intended to demonstrate whether the charcoal filter enhances the recovery time from sevoflurane anesthesia using a semi-closed circuit system. MATERIALS AND METHODS: Thirty healthy patients scheduled for elective surgery under sevoflurane anesthesia were randomly assigned to the charcoal filter or control group. Upon completion of surgery, the end-tidal concentration of sevoflurane was maintained at 2.0 vol%. A charcoal filter was attached to the expiratory limb of the breathing circuit of charcoal filter group subjects. After sevoflurane was discontinued, ventilation was controlled with the same minute volume as the intra-operative period at a fresh gas flow rate of 5 L.min-1 with 100% O2. The elimination kinetics of sevoflurane from end-tidal concentration, Bispectral index and times of eye opening and extubation were obtained. RESULTS: The exponential time constant (tau) of alveolar sevoflurane concentration in the charcoal filter group was significantly shorter than that in the control group (1.7+/-0.5 vs. 2.5+/-1.1 min, p=0.008). The charcoal filter hastened rapid eye opening (11.1+/-3.8 vs. 14.8+/-3.0 min, p=0.007) and extubation (11.9+/-3.9 vs. 15.3+/-3.2 min, p=0.014), compared to the control group. CONCLUSION: A charcoal filter enhances the recovery from sevoflurane anesthesia with a semi-closed rebreathing circuit. PMID- 21623612 TI - A validation study of the Korean version of SPAN. AB - PURPOSE: The SPAN, which is acronym standing for its four components: Startle, Physiological arousal, Anger, and Numbness, is a short post-traumatic stress disorder (PTSD) screening scale. This study sought to develop and validate a Korean version of the SPAN (SPAN-K). MATERIALS AND METHODS: Ninety-three PTSD patients (PTSD group), 73 patients with non-psychotic psychiatric disorders (psychiatric control group), and 88 healthy participants (normal control group) were recruited for this study. Participants completed a variety of psychiatric assessments including the SPAN-K, the Davidson Trauma Scale (DTS), the Clinician Administered PTSD Scale (CAPS), and the State-Trait Anxiety Inventory (STAI). RESULTS: Cronbach's alpha and test-retest reliability values for the SPAN-K were both 0.80. Mean SPAN-K scores were 10.06 for the PTSD group, 4.94 for the psychiatric control group, and 1.42 for the normal control group. With respect to concurrent validity, correlation coefficients were 0.87 for SPAN-K vs. CAPS total scores (p<0.001) and 0.86 for SPAN-K vs. DTS scores (p<0.001). Additionally, correlation coefficients were 0.31 and 0.42 for SPAN-K vs. STAI-S and STAI-T, respectively. Receiver operating characteristic analysis of SPAN-K showed good diagnostic accuracy with an area under the curve (AUC) of 0.87. The SPAN-K showed the highest efficiency at a cutoff score of 7, with a sensitivity of 0.83, a specificity of 0.81, positive predictive value (PPV) of 0.88, and negative predictive value (NPV) of 0.73. CONCLUSION: These results suggest that the SPAN-K had good psychometric properties and may be a useful instrument for rapid screening of PTSD patients. PMID- 21623613 TI - Evaluation of direct immunofluorescence test with PCR for detection of novel influenza A (H1N1) virus during 2009 pandemic. AB - During the 2009 novel influenza (H1N1) pandemic, the sensitivity of direct immunofluorescence assay (DFA) for H1N1 infection was 62% (266/429) of that of the polymerase chain reaction (PCR) test. The sensitivity of the DFA differed significantly with the age of patients: the sensitivity was the highest (71.8%) for patients aged <10 years and the lowest for patients aged >=30 years. The sensitivity of DFA in patients aged >=30 years was 40.7%. Furthermore, the sensitivity (67.3%, 171/254) of DFA was higher for patients who had a high temperature at admission. An increase in the incidence of H1N1 infection did not influence the sensitivity of DFA (62.1% vs. 62%; p=0.984) test, but resulted in a decrease in the negative predictive value, from 92.4% (700/757) to 69.6% (247/355). PCR may be useful as the initial test for diagnosing H1N1 infection in patients aged >=30 years with a normal temperature at presentation. PMID- 21623614 TI - The clinical usefulness of the SD Bioline Influenza Antigen Test(r) for detecting the 2009 influenza A (H1N1) virus. AB - Though the 2009 worldwide influenza A (H1N1) pandemic has been declared to have ended, the influenza virus is expected to continue to circulate from some years as a seasonal influenza. A rapid antigen test (RAT) can aid in rapid diagnosis and allow for early antiviral treatment. We evaluated the clinical usefulness of RAT using SD Bioline Influenza Antigen Test(r) kit to detect the influenza virus, considering various factors. From August 1, 2009 to October 10, 2009, a total of 938 patients who visited the outpatient clinic at Korea University Guro Hospital with influenza-like illnesses were enrolled in the study. Throat or nasopharyngeal swab specimens were obtained from each of the patients. Using these specimens, we evaluated the influenza detection rate by rapid antigen test based on the real-time reverse-transcriptase polymerase chain reaction (rRT-PCR) method. In comparison with rRT-PCR, the sensitivity and specificity of the RAT were 44.0% and 99.9%, respectively. The cyclic threshold values of RAT negative specimens were higher than RAT positive specimens (30.1+/-3.1 vs. 28.3+/-3.9, p=0.031). The sensitivity of the RAT kit was higher in patients who visited clinics within two days of symptom onset (60.4% vs. 11.1%, p=0.026). The results of this study show that the RAT cannot be recommended for general use in all patients with influenza-like illness because of its low sensitivity. The RAT may be used, only in the settings with limited diagnostic resources, for patients who visit a clinic within two days of symptom onse. PMID- 21623615 TI - Clinical features of re-emerging hepatitis A: an analysis of patients hospitalized during an urban epidemic in Korea. AB - From April 2008 to November 2008, many cases of hepatitis A were reported in Seoul and Gyeonggi Province in Korea. Furthermore, the rate of severe or fulminant hepatitis have significantly increased during the latest epidemic (13.4% vs. 5.2%, p=0.044). Therefore, widespread use of vaccine is warranted to reduce the burden of hepatitis A in Korea. PMID- 21623616 TI - Treatment of spontaneous cervical spinal subdural hematoma with methylprednisolone pulse therapy. AB - We report herein a case of hyperacute onset of spontaneous cervical spinal subdural hematoma treated with methylprednisolone pulse therapy that showed good results. A 57-year-old man was admitted for posterior neck pain and paraparesis which occurred an hour ago. MRI revealed a ventral subdural hematoma distributed from the level of C1 down to T3, compressing the spinal cord. Conservative management with methylprednisolone pulse therapy was administered considering the patient's poor general condition. Although emergent surgical decompression is necessary in most cases of spinal subdural hematoma, conservative management with steroid therapy could be effective. PMID- 21623617 TI - Bowel perforation after erlotinib treatment in a patient with non-small cell lung cancer. AB - Erlotinib is accepted as a standard second-line chemotherapeutic agent in patients with non-small cell lung cancer who are refractory or resistant to first line platinum- based chemotherapy. There has been no previous report of bowel perforation with or without gastrointestinal metastases related to erlotinib in patients with non-small cell lung cancer. The exact mechanism of bowel perforation in patients who received erlotinib remains unclear. In this report, we report the first case of enterocutaneous fistula in a female patient with metastatic non-small cell lung cancer 9 months, following medication with erlotinib as second-line chemotherapy. PMID- 21623619 TI - ADAPs worst-case scenario arrives with a vengeance. Waiting list for drugs now tops 6,700 people. PMID- 21623620 TI - Will the safety net hold? New partnerships help. Patient influx 'an administrative nightmare'. PMID- 21623621 TI - Certain cancers increase in pts on long-term ART. Focus should be on screening. PMID- 21623622 TI - Time to boost uptake of HPV vaccine in men. Many think it is only for women. PMID- 21623623 TI - Interim guidance for PrEP in MSM. CDC: Do not expand to other risk groups yet. PMID- 21623624 TI - HIV transmission to HCWs virtually eliminated. But vigilance must be maintained. PMID- 21623625 TI - Higher risk of perinatal HIV transmission in TB mothers. PMID- 21623626 TI - FDA notifications. Genentech issues letter about Fuzeon(r) co-product. PMID- 21623627 TI - Size-dependent lipid content of bovine milk fat globule and membrane phospholipids. AB - The mammary epithelial cell produces unique structures and a range of diversely sized lipid particles from tens of micrometers to less than 1 MUm. The physical, chemical, and biological properties of the differently sized milk fat globules (MFGs) and their complex membranes are not well described. Six size fractions of MFGs were obtained by gravity-based separation and analyzed, and their partial lipidome was determined. The smallest MFGs had a higher concentration of polyunsaturated fatty acids (FAs). The FAs indicative of elongase activity were highest in the smallest MFGs, whereas those FAs indicative of desaturase activity did not differ between size groups. The phosphatidylinositol concentration was highest whereas the phosphatidylserine concentration was lowest in MFGs with an average diameter of 2 MUm. Phosphatidylethanolamine and cholesterol concentrations were highest whereas that of sphingomyelin was lowest in MFGs with an average diameter of 3 MUm. Phosphatidylcholine concentrations did not vary between the size groups. Results suggest that the assembly of milk fat globules that differ in size is not a homogeneous nor random process and that the differences in composition may reflect discrete biosynthetic routes. PMID- 21623628 TI - Li ion diffusion in the anode material Li12Si7: ultrafast quasi-1D diffusion and two distinct fast 3D jump processes separately revealed by 7Li NMR relaxometry. AB - The intermetallic compounds Li(x)Si(y) have attracted considerable interest because of their potential use as anode materials in Li ion batteries. In addition, the crystalline phases in the Li-Si phase diagram turn out to be outstanding model systems for the measurement of fast Li ion diffusion in solids with complex structures. In the present work, the Li self-diffusivity in crystalline Li(12)Si(7) was thoroughly probed by (7)Li NMR spin-lattice relaxation (SLR) measurements. Variable-temperature and -frequency NMR measurements performed in both the laboratory and rotating frames of reference revealed three distinct diffusion processes in Li(12)Si(7). The diffusion process characterized by the highest Li diffusivity seems to be confined to one dimension. It is one of the fastest motions of Li ions in a solid at low temperatures reported to date. The Li jump rates of this hopping process followed Arrhenius behavior; the jump rate was ~10(5) s(-1) at 150 K and reached 10(9) s( 1) at 425 K, indicating an activation energy as low as 0.18 eV. PMID- 21623629 TI - From interfacial ring-opening polymerization to melt processing of cellulose nanowhisker-filled polylactide-based nanocomposites. AB - In the present work, cellulose nanowhiskers (CNWs), extracted from ramie fibers, were incorporated in polylactide (PLA)-based composites. Prior to the blending, PLA chains were chemically grafted on the surface of CNW to enhance the compatibilization between CNW and the hydrophobic polyester matrix. Ring-opening polymerization of l-lactide was initiated from the hydroxyl groups available at the CNW surface to yield CNW-g-PLA nanohybrids. PLA-based nanocomposites were prepared by melt blending to ensure a green concept of the study thereby limiting the use of organic solvents. The influence of PLA-grafted cellulose nanoparticles on the mechanical and thermal properties of the ensuing nanocomposites was deeply investigated. The thermal behavior and mechanical properties of the nanocomposites were determined using differential scanning calorimetry (DSC) and dynamical mechanical and thermal analysis (DMTA), respectively. It was clearly evidenced that the chemical grafting of CNW enhances their compatibility with the polymeric matrix and thus improves the final properties of the nanocomposites. Large modification of the crystalline properties such as the crystallization half time was evidenced according to the nature of the PLA matrix and the content of nanofillers. PMID- 21623630 TI - Synthesis, protein kinase inhibitory potencies, and in vitro antiproliferative activities of meridianin derivatives. AB - The synthesis of new meridianin derivatives is described. The indolic ring system was substituted at the C-4 to C-7 positions either by a bromine atom or by nitro or amino groups. Additionally, an iodine atom or various aryl groups were introduced at the C-5 position of the 2-aminopyrimidine ring. These compounds as well as some of their synthetic intermediates were tested for their kinase inhibitory potencies and for their in vitro antiproliferative activities. We found that this series of compounds is particularly interesting in the development of new inhibitors of DYRK1A and CLK1 kinases. The most effective compounds toward these two kinase families are the 6- and 7-bromo derivatives 30, 33, and 34 that showed more than 45-fold selectivity toward DYRK1A/CLK1 kinases over the other kinases tested. Meridianin derivatives could thus be developed toward potent and selective inhibitors of key RNA splicing regulators and potential therapeutic agents. PMID- 21623631 TI - Structure-activity relationships of a series of analogues of the RFamide-related peptide 26RFa. AB - 26RFa is a new member of the RFamide peptide family that has been identified as the endogenous ligand of the orphan GPCR GPR103. As the C-terminal heptapeptide (26RFa((20-26))) mimics the action of the native peptide on food intake and gonadotropin secretion in rodents, we have synthesized a series of analogues of 26RFa((20-26)) and measured their potency to induce [Ca(2+)](i) mobilization in Galpha(16)-hGPR103-transfected CHO cells. Systematic replacement of each residue by an alanine (Ala scan) and its D-enantiomer (D scan) showed that the last three C-terminal residues were very sensitive to the substitutions while position 23 tolerated rather well both modifications. Most importantly, replacement of Ser(23) by a norvaline led to an analogue, [Nva(23)]26RFa((20-26)), that was 3 fold more potent than the native heptapeptide. These new pharmacological data, by providing the first information regarding the structure-activity relationships of 26RFa analogues, should prove useful for the rational design of potent GPR103 receptor ligands with potential therapeutic application. PMID- 21623632 TI - Electric conductivity-tunable transparent flexible nanowire-filled polymer composites: orientation control of nanowires in a magnetic field. AB - Cobalt compound nanowires were dispersed in a transparent nonconductive polymer film by merely stirring, and the film's transparency and electrical conductivity were examined. This composite film is a unique system in which the average length of the nanowires exceeds the film's thickness. Even in such a system, a percolation threshold existed for the electric conductivity in the direction of the film thickness, and the value was 0.18 vol%. The electric conductivity value changed from ~1 * 10(-12) S/cm to ~1 * 10(-3) S/cm when the volume fraction exceeded the threshold. The electric conductivity apparently followed the percolation model until the volume fraction of the nanowires was about 0.45 vol %. The visible light transmission and electric conductivity of the composite film of about 1 vol % nanowires were 92% and 5 * 10(-3) S/cm, respectively. Moreover, the electric conductivity in the direction parallel to the film surface did not depend on the amount of the dispersed nanowires, and its value was about 1 * 10( 14) S/cm. Even in a weak magnetic field of about 100 mT, the nanowires were aligned in a vertical and parallel direction to the film surface, and the electric conductivity of each aligned composite film was 2.0 * 10(-2) S/cm and 2.1 * 10(-12) S/cm. The relation between the average wire length and the electric conductivity was examined, and the effect of the magnetic alignment on that relation was also examined. PMID- 21623633 TI - Near unity photon-to-electron conversion efficiency of photoelectrochemical cells built on cationic water-soluble porphyrins electrostatically decorated onto thin film nanocrystalline SnO2 surface. AB - Thin transparent SnO(2) films have been surface modified with cationic water soluble porphyrins for photoelectrochemical investigations. Free-base and zinc(II) derivatives of three types of cationic water-soluble porphyrins, (P)M, viz., tetrakis(N-methylpyridyl)porphyrin chloride, (TMPyP)M, tetrakis(trimethylanilinium)porphyrin chloride, (TAP)M, and tetrakis(4'-N methylimidazolyl-phenyl)porphyrin iodide, (TMIP)M, (M = 2H or Zn) are employed. The negative surface charge and the porous structure of SnO(2) facilitated binding of positively charged porphyrins via electrostatic interactions, in addition to strong electronic interactions in the case of (TMPyP)M binding to nanocrystalline SnO(2). The SnO(2)-porphyrin binding in solution was probed by absorption spectroscopy which yielded apparent binding constants in the range of 1.5-2.6 * 10(4) M(-1). Both steady-state and time-resolved fluorescence studies revealed quenching of porphyrin emission upon binding to SnO(2) in water suggesting electron injection from singlet excited porphyrin to SnO(2) conduction band. Addition of LiClO(4) weakened the ion-paired porphyrin-SnO(2) binding as revealed by reversible emission changes. Over 80% of the quenched fluorescence was recovered in the case of (TMPyP)M and (TAP)M compounds but not for (TMIP)M suggesting stronger binding of the latter to SnO(2) surface. Photoelectrochemical studies performed on FTO/SnO(2)/(P)M electrodes revealed incident photon-to current conversion efficiencies (IPCE) up to 91% at the peak maxima for the SnO(2)-dye modified electrodes, with very good on-off switchability. The high IPCE values have been attributed to the strong electrostatic and electronic interactions between the dye, (TMPyP)M and SnO(2) nanoparticles that would facilitate better charge injection from the excited porphyrin to the conduction band of the semiconductor. Electrochemical impedance spectral measurements of electron recombination resistance calculations were supportive of this assignment. PMID- 21623634 TI - Influence of annulene ratio on the electrochemical and spectroscopic properties of methano[10]annulene-thiophene random copolymers. AB - We describe a series of copolymerization studies whereby the nonbenzenoid aromatic methano[10]annulene is incorporated into three different types of random copolymers, two based on polythiophenes (from bithiophene and terthiophene monomers) and one based on poly(ethylene dioxythiophene). Copolymers where the annulene component was in the majority had optical and electrochemical behaviors reminiscent of the annulene homopolymer. In contrast, we found that the annulene influenced polymer electronics at very low feed ratios where the commercial comonomer was in the majority. Copolymerizations are useful approaches to dilute the complex annulene monomers into functional polymers without losing the optoelectronic properties of the annulene homopolymers. These electrochemical results provide important design rules that can be employed for the chemical synthesis of related random copolymers. PMID- 21623635 TI - Three descriptor model sets a high standard for the CSAR-NRC HiQ benchmark. AB - Here we report the results we obtained with a proteochemometric approach for predicting ligand binding free energies of the CSAR-NRC HiQ benchmark data set. Using distance-dependent atom-type pair descriptors in a bagged stepwise multiple linear regression (MLR) model with subsequent complexity reduction we were able to identify three descriptors that can be used to build a very robust regression model for the CSAR-NRC HiQ data set. The model has an R(2)(cv) of 0.55, a MUE(cv) of 1.19, and an RMSE(cv) of 1.49 on the out-of-bag test set. The descriptors selected are the count of protein atoms in a shell between 4.5 A and 6 A around each heavy ligand atom excluding oxygen and phosphorus, the count of sulfur atoms in the vicinity of tryptophan, and the count of aliphatic ligand hydroxy hydrogens. The first two descriptors have a positive sign indicating that they contribute favorably to the binding energy, whereas the count of hydroxy hydrogens contributes unfavorably to the binding free energy observed. The fact that such a simple model can be so effective raises a couple of questions that are addressed in the article. PMID- 21623636 TI - Investigation of sulfonamide, tetracycline, and quinolone antibiotics in vegetable farmland soil in the Pearl River Delta area, southern China. AB - Thirteen antibiotics in soil from vegetable farmlands of the Pearl River Delta, southern China, were investigated. At least three antibiotics were detected in each sample. Six antibiotics including four quinolones, tetracycline, and sulfamethoxazole were detected in >94% of the samples. The total contents of three tetracyclines, eight sulfonamides, and four quinolones were not detected 242.6, 33.3-321.4, and 27.8-1537.4 MUg/kg, respectively. The highest antibiotic concentrations were observed mainly in vegetable farmlands affiliated with livestock farms. Chlortetracycline, sulfameter, and quinolones in some samples exceed the ecotoxic effect trigger value (100 MUg/kg) set by the Steering Committee of Veterinary International Committee on Harmonization. The composition and concentration of antibiotics in soil were correlated with vegetable species. This study has revealed an alarming condition of antibiotics in vegetable farmland soil. Further investigation including environmental fate, plant uptake, and human exposure to antibiotics by plant-derived food should be conducted. PMID- 21623637 TI - Quantitative chemical imaging of element diffusion into heterogeneous media using laser ablation inductively coupled plasma mass spectrometry, synchrotron micro-X ray fluorescence, and extended X-ray absorption fine structure spectroscopy. AB - Quantitative chemical imaging of trace elements in heterogeneous media is important for the fundamental understanding of a broad range of chemical and physical processes. The primary aim of this study was to develop an analytical methodology for quantitative high spatial resolution chemical imaging based on the complementary use of independent microanalytical techniques. The selected scientific case study is focused on high spatially resolved quantitative imaging of major elements, minor elements, and a trace element (Cs) in Opalinus clay, which has been proposed as the host rock for high-level radioactive waste repositories. Laser ablation inductively coupled plasma mass spectrometry (LA ICPMS), providing quantitative chemical information, and synchrotron radiation based micro-X-ray fluorescence (SR-microXRF), providing high spatial resolution images, were applied to study Cs migration into Opalinus clay rock. The results indicate that combining the outputs achievable by the two independent techniques enhances the imaging capabilities significantly. The qualitative high resolution image of SR-microXRF is in good agreement with the quantitative image recorded with lower spatial resolution by LA-ICPMS. Combining both techniques, it was possible to determine that the Opalinus clay sample contains two distinct domains: (i) a clay mineral rich domain and (ii) a calcium carbonate dominated domain. The two domains are separated by sharp boundaries. The spatial Cs distribution is highly correlated to the distribution of the clay. Furthermore, extended X-ray absorption fine structure analysis indicates that the trace element Cs preferentially migrates into clay interlayers rather than into the calcite domain, which complements the results acquired by LA-ICPMS and SR microXRF. By using complementary techniques, the quantification robustness was improved to quantitative micrometer spatial resolution. Such quantitative, microscale chemical images allow a more detailed understanding of the chemical reactive transport process into and within heterogeneous media to be gained. PMID- 21623638 TI - Antimicrobial and anti-thrombogenic features combined in hydrophilic surface coatings for skin-penetrating catheters. Synergy of co-embedded silver particles and heparin. AB - Percutaneous (skin-penetrating) catheters such as central venous catheters (CVCs), are used ubiquitously in the treatment of critically ill patients, although it is known that the risks for serious complications, particularly bloodstream infection and thromboembolism, are high. Materials science and engineering offer important new perspectives regarding further improvement of CVCs. A promising approach is the use of synthetic biocompatible hydrogel coatings with both silver particles and heparin embedded therein. Such formulations combine the well-known broad-spectrum antimicrobial features of silver with the anticoagulant activity of immobilized heparin. Previous work revealed that heparin augments antimicrobial activity of silver, while maintaining its anticoagulant function. This study set out to investigate the synergy of heparin and silver in more detail. Exit-challenge tests, experiments on bacterial killing and adherence, as well as in vitro challenge tests with three Staphylococcus aureus strains (one reference strain, and two clinical isolates) consistently showed the synergistic effect. In addition, the impact of changing the coating's hydrophilicity, and changing the silver concentration in the coatings, were examined. The experimental results, taken together and combined with data from the literature, point out that synergy of heparin and silver is best explained by binding of Ag(+) ions to heparin within the swollen coating, followed by release of heparin-Ag(+) complexes upon immersion of the coatings in an aqueous environment such as blood. Possible implications of this work regarding the development of improved/safer CVCs are briefly discussed. PMID- 21623639 TI - Probing DNA's interstrand orientation with gold nanoparticles. AB - The interstrand orientation of a DNA duplex plays a pivotal role in its biological and chemical functions. Therefore, developing an efficient way to determine (control and monitor) the parallel or antiparallel conformation of a DNA duplex is of great significance, which, however, remains a big challenge under some circumstances. In this work, we demonstrate that gold nanoparticles tagged on DNA are especially useful in trapping and detecting a special interstrand orientation of a DNA double helix, based on inherent electrostatic and steric repulsions between nanoparticles which will affect their self-assembly into a large structure. More importantly, some of the conformations revealed by the gold nanoparticle assay may even not be thermodynamically preferred and thus will be hard to detect using currently available methods. This simple, straightforward, and efficient methodology capable of dictating and probing a special DNA duplex structure provides a useful tool for conformational analyses and functional explorations of biomolecules as well as biophysical and nanobiomedical research. PMID- 21623640 TI - Bisoxazolidine-catalyzed enantioselective Reformatsky reaction. AB - A readily available chiral bisoxazolidine catalyzes the asymmetric Reformatsky reaction between ethyl iodoacetate and aldehydes. In the presence of 10 mol % of the ligand, dimethylzinc, and air, this method produces ethyl 3-hydroxy-3-(4 aryl)propanoates in high yields and in 75 to 80% ee at room temperature within 1 h. In contrast to aromatic substrates, relatively low ee's are obtained with aliphatic aldehydes. PMID- 21623641 TI - Transdermal rivastigmine: management of cutaneous adverse events and review of the literature. AB - Alzheimer's disease is a chronic neurodegenerative disorder resulting in part from the degeneration of cholinergic neurons in the brain. Rivastigmine, a cholinesterase inhibitor, is commonly used as a treatment for dementia due to its ability to moderate cholinergic neurotransmission; however, treatment with oral rivastigmine can lead to gastrointestinal adverse effects such as nausea and vomiting. Transdermal administration of rivastigmine can minimize these adverse effects by providing continuous delivery of the medication, while maintaining the effectiveness of the oral treatment. While the transdermal form of rivastigmine has been found to have fewer systemic adverse effects compared with the oral form, cutaneous reactions, such as contact dermatitis, can lead to discontinuation of the drug in its transdermal form. Lack of patient compliance with regard to applying the patch to the designated site, applying the patch for the correct length of time or rotating patch application sites increases the risk of cutaneous adverse reactions. This article outlines the diagnosis and management of irritant contact dermatitis and allergic contact dermatitis secondary to transdermal rivastigmine. The large majority of reactions to transdermal patches are of an irritant type, which can be diagnosed clinically by the presence of a pruritic, erythematous, eczematous plaque strictly confined to the borders of the patch. In contrast, an allergic reaction can be differentiated by the presence of vesicles and/or oedema, erythema beyond the boundaries of the transdermal patch and lack of improvement of the lesion 48 hours after removal of the offending treatment. By encouraging the patient to follow a regular rotation schedule for the patch, and using lipid-based emollients for irritant dermatitis and pre- and post-treatment topical corticosteroids for allergic dermatitis, cutaneous reactions can often be alleviated and patients can continue with their medication regimen. Other simple changes to a patient's treatment routine, including minimizing the use of harsh soaps, avoiding recently shaven or damaged areas of skin and carefully removing the patch after use, can help to further decrease the risk of dermatitis development. PMID- 21623642 TI - Role of irbesartan in prevention of post-coronary artery bypass graft atrial fibrillation. AB - BACKGROUND AND OBJECTIVE: Atrial fibrillation (AF) is a common complication of cardiothoracic surgery (CTS). Existing evidence about the potential protective role of angiotensin II type 1 receptor antagonists (angiotensin receptor blockers [ARBs]) for post-CTS AF has been limited and conflicting. In this single-blind, open-label, randomized prospective pilot study, we evaluated the potential protective role of irbesartan (an ARB) in post-coronary artery bypass graft (CABG) AF. METHODS: A total of 100 consecutive patients undergoing CABG were randomly assigned to irbesartan (n = 50) versus no irbesartan (n = 50) for 5 days prior to the scheduled surgery. Data were collected for imaging studies, laboratory values, and peri-operative details. Patients were monitored post operatively for in-hospital AF episodes. Unadjusted and adjusted logistic regression analysis was performed to assess the effect of irbesartan on the incidence of post-CABG AF. RESULTS: A total of 14 patients developed AF during their post-operative hospital stay. The incidence of AF in patients who received irbesartan was 6% (n = 3) compared with 22% (n = 11) in patients who did not receive irbesartan (p = 0.021). Univariate logistic regression analysis identified irbesartan and age as statistically significant variables. An adjusted multivariate logistic model identified irbesartan as an important protective factor against development of post-CABG AF (adjusted odds ratio [OR] 0.20; 95% confidence interval [CI] 0.04, 0.94; p = 0.04). Increasing age (adjusted OR 1.09, 95% CI 1.01, 1.17; p = 0.03) was also identified as an independent risk factor for development of post-CABG AF. CONCLUSION: Pretreatment with irbesartan tends to have a significant protective effect against the occurrence of AF during the post-operative period in patients undergoing CABG. PMID- 21623643 TI - Ambrisentan. AB - Ambrisentan, an orally active, highly selective antagonist of the endothelin-1 type A receptor, is indicated for the treatment of pulmonary arterial hypertension (PAH). It has a low potential for drug-drug interactions and requires only once-daily administration. Three months' treatment with ambrisentan 2.5-10 mg/day significantly improved exercise capacity, as determined by the distance walked in 6 minutes (6MWD; primary outcome measure), compared with placebo in two double-blind, multicenter studies in patients with PAH (ARIES-1 [n = 202] and -2 [n = 192]). A decrease in dyspnea and a delay in clinical worsening were among the improvements in secondary outcomes generally observed with ambrisentan versus placebo. In ARIES-E, a 2-year extension of ARIES-1 and -2, approved dosages of ambrisentan (5 and 10 mg/day) were associated with a sustained improvement in 6MWD, a generally sustained improvement in dyspnea, and a low risk of clinical worsening and of death. Six months' treatment with ambrisentan 5 mg/day significantly improved 6MWD (primary outcome measure) and dyspnea relative to baseline in an open-label, non-comparative, multicenter study in a diverse population of patients with PAH or non-PAH forms of pulmonary hypertension (ARIES-3 [n = 224]). Ambrisentan was associated with a low risk of clinical worsening and of death. Ambrisentan treatment was generally well tolerated in the various ARIES trials. All available pre-registration and post marketing data indicate the drug poses only a very low risk of liver injury; the 'black box' warning regarding potential liver injury has been removed from the US prescribing information for ambrisentan. PMID- 21623644 TI - The influenza pandemic of 2009: lessons and implications. AB - Influenza is a moving target, which evolves in unexpected directions and is recurrent annually. The 2009 influenza A/H1N1 pandemic virus was unlike the 2009 seasonal virus strains and originated in pigs prior to infecting humans. Three strains of viruses gave rise to the pandemic virus by antigenic shift, reassortment, and recombination, which occurred in pigs as 'mixing vessels'. The three strains of viruses had originally been derived from birds, pigs, and humans. The influenza hemagglutinin (HA) and neuraminidase (NA) external proteins are used to categorize and group influenza viruses. The internal proteins (PB1, PB1-F2, PB2, PA, NP, M, and NS) are involved in the pathogenesis of influenza infection. A major difference between the 1918 and 2009 pandemic viruses is the lack of the pathogenic protein PB1-F2 in the 2009 pandemic strains, which was present in the more virulent 1918 pandemic strains. We provide an overview of influenza infection since 1847 and the advent of influenza vaccination since 1944. Vaccines and chemotherapy help reduce the spread of influenza, reduce morbidity and mortality, and are utilized by the global rapid-response organizations associated with the WHO. Immediate identification of impending epidemic and pandemic strains, as well as sustained vigilance and collaboration, demonstrate continued success in combating influenza. PMID- 21623646 TI - Attitudes of health care professionals toward pharmacogenetic testing. AB - BACKGROUND: Pharmacogenetics has emerged as a new tool for the optimization of drug therapy. Although the pharmacogenetics concept was first recognized at least 50 years ago, clinical testing to determine pharmacogenetic traits is still relatively rare, and many hurdles are markedly slowing its development. There is a lot of literature and speculation about potential ethical challenges in genetic and pharmacogenetic testing, yet few researchers have actually examined the attitudes of health care professionals regarding the clinical application of these tests. OBJECTIVE: In this article, we aim to review the current literature on health care professionals' perceptions of the role of pharmacogenetic data and describe the attitudes of medical students when faced with a clinical pharmacogenetic testing scenario. METHODS: A group of 59 third-year medical students from the American University of Beirut Medical Center were asked to answer a questionnaire about pharmacogenetic testing after being exposed to a clinical scenario of a patient who was diagnosed with mild Alzheimer Disease (AD) and hence was a candidate for therapy with one of the acetylcholinesterase (AChE) inhibitors. RESULTS: The students indicated that they would respect patients' confidentiality and inform them about the test results and therapeutic plan, but they would not be as open about bad prognoses. They did not agree on the therapeutic plan that would follow a pharmacogenetic test result and were uncertain about potential patient discrimination in insurability. CONCLUSION: Our and others' findings demonstrate the existence and seriousness of several challenges pertaining to pharmacogenetic applications in the clinical setting. Further training and education are needed for health care professionals, since they are the ones who will most probably request these tests in the near future. PMID- 21623648 TI - Is centralization in emergency rural medicine always right? Lessons learned from two cases of decompression sickness. AB - The cases of two patients with decompression sickness (DS) are described to add to the discussion about whether centralization, especially when accompanied by air-medical transport, is always justifiable in island emergency medicine. One patient received hyperbaric oxygen (HBO) treatment on another island after island to-island transfer by boat; the other received HBO treatment on a ship that was anchored, by chance, close to the island where he became ill. Both cases had a good outcome. Island-to-island transport and within-island treatment, rather than island-to-urban-center transport, was effective, indicating that treatment centralization may not be the most effective protocol all cases. A DS treatment strategy is proposed for use in this geographic area; however, DS occurring on remote islands highlights the wider issue of the centralization of health services. PMID- 21623645 TI - Blood-based protein biomarkers for diagnosis and classification of neurodegenerative diseases: current progress and clinical potential. AB - Biomarker research is a rapidly advancing field in medicine. Recent advances in genomic, genetic, epigenetic, neuroscientific, proteomic, and metabolomic knowledge and technologies have opened the way to thriving research. In the most general sense, a biomarker refers to any useful characteristic that can be measured and used as an indicator of a normal biologic process, a pathogenic process, or a pharmacologic response to a therapeutic agent. Despite the extensive resources concentrated on this area, there are very few biomarkers currently available that qualify and are satisfactorily validated for mental disorders, and there is still a major lack of biomarkers for typifying neurodegenerative disorders such as Alzheimer's disease and Parkinson's disease. This article provides an overview of this field of research and focuses on recent advances in biomarker research in Alzheimer's disease and Parkinson's disease. PMID- 21623649 TI - The vitamin B12 absorption test, CobaSorb, identifies patients not requiring vitamin B12 injection therapy. AB - BACKGROUND: Treatment with vitamin B12 has virtually no side effects; however, life-long treatment is inconvenient for the patient and constitutes a cost for society. OBJECTIVE: To investigate whether vitamin B12 injection treatment reflects the actual need for treatment or whether some patients are treated unnecessarily with vitamin B12 injections. MATERIAL AND METHODS: A prospective intervention study was conducted among nine general practitioners in Western Sealand County, Denmark. Forty-four patients older than 18 years who had received injection therapy with vitamin B12 for a median of eight years (range 1-26 years) were included. After discontinuation of vitamin B12 injections, blood samples were analysed monthly for hemoglobin, cobalamin, holotranscobalamin, homocysteine and methylmalonic acid. The capacity to absorb vitamin B12 was examined after a median of 13 months (range 5-32 months) by measurement of holotranscobalamin or cyanocobalamin on transcobalamin before and after 1 and 2 days intake of 3 * 9 MUg of vitamin B12. Patients unable to absorb the vitamin continued treatment with vitamin B12 injection. The remaining patients participated in a follow-up study receiving 9 MUg oral vitamin B12 daily or no vitamin B12 substitution. RESULTS: Of the 44 patients studied, 35 patients were able to absorb vitamin B12. None of the patients included in the follow-up study showed biochemical signs of vitamin B12 deficiency by the end of the study. CONCLUSION: Our results suggest that the capacity for absorbing vitamin B12 should be examined prior to the choice of treatment. PMID- 21623650 TI - The influence of informant characteristics on the reliability of family history interviews. AB - Family history interviews are widely used in psychiatric research, as well as in genetic and twin studies, and provide a way to collect family history information quickly and economically. To obtain a valid assessment of family history, it is important to investigate which family member will be able to provide accurate information. Previous research shows that the validity of family history reporting can be influenced by characteristics of the informant, such as age, gender and personal history of psychiatric disorder. The aim of this study was to investigate the role of a subject's position in a pedigree on the validity of data collection. Family history data on diabetes and psychiatric disorders were collected in three generations of 33 families by interviewing both an index subject (3rd generation) and his or her mother (2nd generation). Mothers were shown to report higher rates of diabetes and psychiatric disorder in the family compared to the index subjects. There was no significant difference in the disease rate reported by male and female index subject. Mothers who experienced a depressive episode indicated significantly more family members as having a psychiatric disorder than mothers who never experienced such an episode. This could be explained by the presence of informant bias, but may also result from the fact that depression is a heritable disorder and is therefore actually more prevalent in these families. Our findings suggest that family interview data should be collected by interviewing subjects who have a central position in the pedigree and can therefore provide information on his/her own generation, the previous and the next. In addition, psychiatric status of the informant should be carefully addressed. PMID- 21623651 TI - Statistical approach to prenatal zygosity assessment following a decade of molecular aneuploidy screening. AB - In twin pregnancy studies, molecular genetic techniques have rarely been used to determine zygosity, despite their known precision and accuracy. The present work aimed to assess the power of discrimination in zygosity assessment, using a set of microsatellite markers that were routinely used for aneuploidy screening by multiplex-PCR in a prenatal context. Rapid aneuploidy screening using a group of 20 microsatellite markers (STRs) located on chromosomes 13, 18, 21 and X has been performed in our lab for over 10 years, with a total of approximately 1,500 samples studied to date. A retrospective analysis of the 257 prenatal samples from multiple pregnancies was carried out. A subset of 14 cases presenting theoretical monozygosity were re-evaluated by the use of biostatistics tools accessed via the ZygProb website. Further monozygosity determination relative to dizygosity was calculated, given an estimated overall error value of 0.093%. The results show that monozygosity had been correctly determined in all our previously studied twins. This work demonstrates that accurate zygosity assessment can be achieved with the same STRs applied in aneuploidy screening with a high power of discrimination and a matching probability of over 99.999999%. PMID- 21623652 TI - SNP sets and reading ability: testing confirmation of a 10-SNP set in a population sample. AB - A set of 10 SNPs associated with reading ability in 7-year-olds was reported based on initial pooled analyses of 100K SNP chip data, with follow-up testing stages using pooling and individual testing. Here we examine this association in an adolescent population sample of Australian twins and siblings (N = 1177) aged 12 to 25 years. One (rs1842129) of the 10 SNPs approached significance (P = .05) but no support was found for the remaining 9 SNPs or the SNP set itself. Results indicate that these SNPs are not associated with reading ability in an Australian population. The results are interpreted as supporting use of much larger SNP sets in common disorders where effects are small. PMID- 21623653 TI - A behavior genetic study of the connection between social values and personality. AB - The present research examined the extent to which relations between social values and personality are due to shared genetic or environmental factors. Using the Rokeach (1973) Value Survey and a scoring key from Schwartz and Bilsky (1990), seven value scores (enjoyment, achievement, self-direction, maturity, prosocial, security, and restrictive conformity) were derived in a sample of twins. As expected, all of the value scales were found to have a significant genetic component, with values ranging from 36% for enjoyment to 63% for prosocial, and there were numerous significant phenotypic correlations found between the value scales and personality scores. Most important, bivariate genetic analyses revealed that some of these phenotypic correlations could be attributed to common genetic or environmental factors. PMID- 21623654 TI - An epidemiological survey of post-coital psychological symptoms in a UK population sample of female twins. AB - Postcoital psychological symptoms (PPS) is a virtually unexplored phenomenon in the female population even though women frequently complain about irritability and motiveless crying after intercourse and/or orgasm. The aim of this study was to explore the epidemiology and genetic influences of PPS in a UK population sample of women. 1,489 unselected female twins aged 18-85 completed questions on recent and persistent PPS and potential risk factors. Standard methods of quantitative genetic analysis were used to model latent genetic and environmental factors influencing variation in PPS. For identification of potential risk factors, regression analyses were conducted. Phenotypic variation in PPS was explored using a genetic variance component analysis (VCA) approach. We found 3.7% of women reported suffering from recent PPS and 7.7% from persistent PPS. Relationship satisfaction and experience of abuse were found to be independently associated with recent (OR 4.5, 95% CI 4.13-4.87 and OR 1.3, 95% CI 1.02-1.34, respectively) and persistent PPS (OR 2.53, 95% CI 2.17-2.81 and OR 1.16, 95% CI 1.09-1.26, respectively). VCA revealed that phenotypic variance was best explained by an additive genetic (AE) model, ascribing 28% (for recent PPS) and 26% (for persistent PPS) of phenotypic variance to additive genetic effects, with the rest being a result of individual experiences and random measurement error. To our knowledge, this is the first and largest study investigating the epidemiology of PPS. It seems that the most important targets for intervention and prevention occur outside of the family, such as relationship quality and satisfaction, and history of abuse. PMID- 21623655 TI - Subjective wellbeing and longevity: a co-twin control study. AB - Mental health is increasingly defined not only by the absence of illness but by the presence of subjective well-being (SWB). Previous cohort studies have consistently shown that indicators of SWB predict favorable life outcomes, including better mental and somatic health, and longevity. The favorable effects associated with SWB have prompted new research aimed at raising happiness and wellbeing through individual interventions and public health initiatives. Standard observational studies of individual-level associations, however, are subject to potential confounding of exposure and outcome by shared genes and environment. The present study explored the association between SWB and increased longevity, using twin pair analyses to determine whether the association is consistent with causality or is due to genetic or environmental confounding. The study sample of 3,966 twins aged 70 or older, followed for a median time period of 9 years, was drawn from the population-based Longitudinal Study of Aging Danish Twins (LSADT). The association between SWB, operationalized as affect and life satisfaction, and all-cause mortality risk was examined using between individual and within-pair survival analyses. As expected, at the individual level, SWB predicted increased longevity. Exposure effects were also present in unadjusted and adjusted within-pair analyses of 400 dizygotic (DZ) pairs and 274 monozygotic (MZ) pairs, indicating that SWB is associated with increased longevity independent of familial factors of genes and shared environment. PMID- 21623656 TI - Genome-wide linkage analysis of hemodynamic parameters under mental and physical stress in extended Omani Arab pedigrees: the Oman Family Study. AB - BACKGROUND: We performed a genome-wide scan in a homogeneous Arab population to identify genomic regions linked to blood pressure (BP) and its intermediate phenotypes during mental and physical stress tests. METHODS: The Oman Family Study subjects (N = 1277) were recruited from five extended families of ~10 generations. Hemodynamic phenotypes were computed from beat-to-beat BP, electrocardiography and impedance cardiography. Multi-point linkage was performed for resting, mental (word conflict test, WCT) and cold pressor (CPT) stress and their reactivity scores (s), using variance components decomposition-based methods implemented in SOLAR. RESULTS: Genome-wide scans for BP phenotypes identified quantitative trait loci (QTLs) with significant evidence of linkage on chromosomes 1 and 12 for WCT-linked cardiac output (LOD = 3.1) and systolic BP (LOD = 3.5). Evidence for suggestive linkage for WCT was found on chromosomes 3, 17 and 1 for heart rate (LOD = 2.3), DBP (LOD = 2.4) and left ventricular ejection time (LVET), respectively. For ?WCT, suggestive QTLs were detected for CO on chr11 (LOD = 2.5), LVET on chr3 (LOD = 2.0) and EDI on chr9 (LOD = 2.1). For CPT, suggestive QTLs for HR and LVET shared the same region on chr22 (LOD 2.3 and 2.8, respectively) and on chr9 (LOD = 2.3) for SBP, chr7 (LOD = 2.4) for SV and chr19 (LOD = 2.6) for CO. For ?CPT, CO and TPR top signals were detected on chr15 and 10 (LOD; 2.40, 2.08) respectively. CONCLUSION: Mental stress revealed the largest number of significant and suggestive loci for normal BP reported to date. The study of BP and its intermediate phenotypes under mental and physical stress may help reveal the genes involved in the pathogenesis of essential hypertension. PMID- 21623657 TI - Twin-singleton differences in neonatal brain structure. AB - Twin studies suggest that global and regional brain volumes are highly heritable. However, estimates of heritability vary across development. Given that all twin studies are open to the potential criticism of non-generalizability due to differences in intrauterine environment between twins and singletons, these age effects may reflect the influence of perinatal environmental factors, which are unique to twins and which may be especially evident early in life. To address this question, we compared brain volumes and the relationship of brain volumes to gestational age in 136 singletons (67 male, 69 female) and 154 twins (75 male, 79 female; 82 DZ, 72 MZ) who had received high resolution MRI scans of the brain in the first month of life. Intracranial volume, total white matter, and ventricle volumes did not differ between twins and singletons. However, cerebrospinal fluid and frontal white matter volume was greater in twins compared to singletons. While gray matter volumes at MRI did not differ between groups, the slope of the relationship between total and cortical gray matter and gestational age at the MRI scan was steeper in MZ twins compared to DZ twins. Post-hoc analyses suggested that gray matter development is delayed in MZ twins in utero and that they experience 'catch-up' growth in the first month of life. These differences should be taken into account when interpreting and designing studies in the early postnatal period. PMID- 21623658 TI - Resemblance of tongue anatomy in twins. AB - This study compared the anatomical features of the tongue in nine pairs of twins six monozygotic and three dizygotic. The aim of the project was to determine if tongues, like any other anatomical structure, could be used to reliably predict relatedness given that tongue shape, presentation and surface can be influenced by environment. Using the method of forced choice, 30 subjects were asked to match the photographs of tongues from twins. Our data indicate that, based on visual assessment, monozygotic twins have highly similar tongues (60% matches); similarly, dizygotic twins were matched 31% of the time, which is a higher probability than would be expected from random selection. This study should help identify baseline and control data in future behavioral studies of taste, which has a genetic basis. PMID- 21623659 TI - Expectant management in twin pregnancies with discordant structural fetal anomalies. AB - OBJECTIVE: Routine obstetric ultrasound increasingly leads to the detection of structural fetal anomalies. In twin pregnancies with one anomalous twin, counseling on management strategies is complicated. PATIENTS AND METHODS: Twin pregnancies (n = 212) were referred to a tertiary center between January 2007 and July 2009. In a retrospective analysis, twins discordant for a structural fetal anomaly were compared to twins without anomalies in the prenatal ultrasound. Outcome parameters were survival and gestational age at birth. RESULTS: Anomalies were seen in at least one fetus of 30 twin pairs. The two pregnancies in which the anomalies were concordant were terminated. Selective feticide was performed in three cases of major but non-lethal anomalies in dichorionic twins. The remaining 25 cases were managed expectantly. In three of these cases, spontaneous fetal demise of the affected fetus was observed. In five cases with major (lethal) anomalies, the pregnant women and their partners opted for non intervention comfort care after birth for the affected fetus. Median gestational age at delivery was 257 days for twins without structural anomalies and was 254 days (n = 22) for twins with one anomalous fetus. This was not significantly different (Mann Whitney U, p = .69). Again, no difference was found for median gestational age at delivery in normal vs. discordant anomalous dichorionic twins if subdivided for chorionicity (Mann Whitney U, p = .68). CONCLUSION: In this cohort we describe the request for expectant management by pregnant women and their partners of those twins discordant for major (lethal) anomalies. Expectant management was not associated with increased risk of premature delivery. Fetocide was only opted for in a small number of cases with severe but non-lethal anomalies in dichorionic twins. PMID- 21623660 TI - Reunited twins: spouse relations / twin research reports / timely topics. AB - The present article explores the social attraction that may evolve on the part of reared apart twins' spouses toward their brothers- and sisters-in-law, that is, their spouses' newly found co-twin. This topic was inspired by the fascinating story of monozygotic reared apart (MZA) twins who were reunited in Perth, Australia in 2008, at age 50. It is followed by brief reviews of twin research articles concerning divorce rates among mothers of multiples and X-chromosome inactivation in MZ female twin pairs. The final section presents informative human interest stories involving twins. PMID- 21623661 TI - Toll-like receptor 3 gene expression in Egyptian patients with glomerulonephritis and hepatitis C virus infection. AB - BACKGROUND: Hepatitis C virus (HCV) infection is associated with both chronic liver disorders and several extrahepatic manifestations including renal disease. Toll-like receptor 3 (TLR3), a component of the innate immune system, is a pathogen recognition receptor that recognizes viral double-stranded RNA. OBJECTIVE: This work investigated TLR3 expression in peripheral blood mononuclear cells from HCV-positive patients with glomerulonephritis. METHODS: One hundred and thirty patients with glomerulonephritis were initially enrolled in the study. After exclusion of 52 patients with secondary glomerulonephritis, 78 patients were screened for HCV infection. TLR3 expression in peripheral blood mononuclear cells was studied in 46 patients with HCV-positive glomerulonephritis and 32 patients with HCV-negative glomerulonephritis using a real-time PCR comparative quantitation approach and results were compared to a control group of 20 healthy subjects. RESULTS: TLR3 expression was significantly higher in patients with HCV positive glomerulonephritis than in HCV negative patients and controls (p < 0.0001). TLR3 expression correlated positively with HCV viral load, interleukin 1beta, serum creatinine and inversely with creatinine clearance in patients with HCV-positive glomerulonephritis. CONCLUSION: This work shows that TLR3 expression is upregulated in HCV-positive patients with glomerulonephritis. Overexpression is associated with reduced renal function and increased interleukin 1beta level. PMID- 21623662 TI - Anti-RNA polymerase III antibodies are associated with scleroderma renal crisis in a French cohort. PMID- 21623663 TI - Common MEFV mutations in Iranian Azeri Turkish patients with Behcet's disease. AB - OBJECTIVE: Behcet's disease (BD) is an inflammatory disorder of unknown cause with higher prevalence along the ancient Silk Road. BD shares epidemiological and clinical features with familial Mediterranean fever (FMF). Moreover, association of BD and certain MEFV gene mutations has been described in recent decades. We studied the role of MEFV mutations in Iranian Azeri Turkish patients with BD. METHODS: Fifty-three BD patients who met the International Study Group criteria for BD were analysed for five common MEFV mutations (M694V, V726A, M680I, M694I, and E148Q) using amplification refractory mutation system and polymerase chain reaction (PCR) restriction-digestion testing methods. A cohort of 200 healthy Azeri Turkish individuals who had been previously genotyped regarding the five common MEFV mutations served as the control group. RESULTS: Eighteen patients were found to carry a single MEFV mutation and one additional patient was compound heterozygote. There was a statistically significant difference between the patient group and ethnically matched healthy individuals regarding M694V and M680I mutations (p = 0.01 and p = 0.04, respectively). Both BD groups (carriers and non-carriers of MEFV mutations) were similar in their clinical symptoms. CONCLUSION: Definite MEFV mutations seem to be a susceptibility factor for BD in our cohort of Iranian Azeri Turkish patients. PMID- 21623664 TI - Autonomic dysfunction in the early stage of ALS with bulbar involvement. AB - Our objective was to assess the autonomic function of ALS patients with and without bulbar signs to characterize dysautonomia in ALS disease. Standard autonomic tests and spectral analysis of heart rate variability (HRV), reflecting changes in the sympathovagal balance, were examined in 33 ALS patients (14 with bulbar signs) and 30 controls. Results showed that in the supine position, ALS patients had significantly lower total power and absolute values of high frequency power indicating a depressed sinus arrhythmia. Patients with bulbar signs showed more marked autonomic alterations at rest. Tilting did not induce the expected increase in low-frequency and decrease in high-frequency power of HRV in all patients. No correlation was found between autonomic tests and clinical parameters. Our findings suggest an early subclinical involvement of the autonomic system in ALS, particularly affecting parasympathetic cardiac control. Patients with prominent bulbar signs show a more severe autonomic dysfunction under resting conditions. PMID- 21623666 TI - Long-term outcome of lung transplantation in a patient with the anti-synthetase syndrome. PMID- 21623665 TI - Talampanel reduces the level of motoneuronal calcium in transgenic mutant SOD1 mice only if applied presymptomatically. AB - We tested the efficacy of treatment with talampanel in a mutant SOD1 mouse model of ALS by measuring intracellular calcium levels and loss of spinal motor neurons. We intended to mimic the clinical study; hence, treatment was started when the clinical symptoms were already present. The data were compared with the results of similar treatment started at a presymptomatic stage. Transgenic and wild-type mice were treated either with talampanel or with vehicle, starting in presymptomatic or symptomatic stages. The density of motor neurons was determined by the physical disector, and their intracellular calcium level was assayed electron microscopically. Results showed that motor neurons in the SOD1 mice exhibited an elevated calcium level, which could be reduced, but not restored, with talampanel only when the treatment was started presymptomatically. Treatment in either presymptomatic or symptomatic stages failed to rescue the motor neurons. We conclude that talampanel reduces motoneuronal calcium in a mouse model of ALS, but its efficacy declines as the disease progresses, suggesting that medication initiation in the earlier stages of the disease might be more effective. PMID- 21623667 TI - Hearing voices: a role of interhemispheric auditory connectivity? AB - OBJECTIVES: Auditory verbal hallucinations (AVH) are among the most common symptoms in schizophrenia. Earlier studies suggest changes in the structural connectivity of auditory areas involved in the pathophysiology of auditory hallucinations. Combining diffusion tensor imaging (DTI) and fibre tractography provides a unique opportunity to visualize and quantify entire fibre bundles. METHODS: Fibre tracts connecting homotopic auditory areas via the corpus callosum were identified with DTI in ten first episode paranoid schizophrenia patients and ten healthy controls. Regions of interest were drawn manually, to guide tractography, and fractional anisotropy (FA) - a measure of fibre integrity - was calculated and averaged over the entire tract for each subject. RESULTS: There was no difference in the FA of the interhemispheric auditory fibres between schizophrenic patients and healthy controls. However, the subgroup of patients hearing conversing voices showed increased FA relative to patients without these symptoms (P = 0.047) and trendwise increased FA relative to healthy controls (P = 0.066). In addition, a trendwise correlation between FA values and AVH symptoms (P = 0.089) was found. CONCLUSIONS: Our findings suggest that in addition to local deficits in the left auditory cortex and disturbed fronto-temporal connectivity, the interhemispheric auditory pathway might be involved in the pathogenesis of AVH. PMID- 21623668 TI - Safety and efficacy of repetitive transcranial magnetic stimulation in the treatment of obsessive-compulsive disorder: a review. AB - OBJECTIVES: Obsessive-compulsive disorder (OCD) is a chronic, often severe, neuropsychiatric disorder leading to a dramatic impairment in interpersonal and occupational functions. rTMS has been tried out in several studies in patients with OCD with different characteristics. In this paper, we review the safety and efficacy of rTMS in the treatment of mostly severe resistant OCD. METHODS: A review of the English literature from 1966 to 2010 pertaining to rTMS in the treatment of OCD was conducted using MEDLINE by selectively entering the search terms "transcranial magnetic stimulation", "repetitive transcranial magnetic stimulation", "obsessive-compulsive disorder" and "OCD". Twelve studies including open and randomized, sham-controlled trials were included in this review. RESULTS: Although available data about the use of rTMS in OCD treatment are quite heterogeneous in terms of sample size, study design, stimulus parameters used and stimulation areas targeted, promising findings regarding rTMS efficacy appeared for two structures based on recent controlled studies: the supplementary motor area and the orbitofrontal cortex. On the other hand, rTMS of the dorsolateral prefrontal cortex is not significantly effective when compared to sham rTMS. CONCLUSIONS: Three target areas have already been selected of which the supplementary motor area in particular and the orbitofrontal cortex seem to be the most promising in terms of potential efficacy and could more accurately be targeted with the help of neuronavigational techniques. Larger randomized controlled trials should be conducted in order to better clarify the therapeutic role of rTMS in OCD. PMID- 21623669 TI - Epidermal growth factor, basic fibroblast growth factor and platelet-derived growth factor-bb can substitute for fetal bovine serum and compete with human platelet-rich plasma in the ex vivo expansion of mesenchymal stromal cells derived from adipose tissue. AB - BACKGROUND AIMS: Human mesenchymal stromal cells (MSC) are multipotent cells possessing self-renewal capacity, long-term viability and multilineage potential. We analyzed the effect of four different medium supplements on the expansion and differentiation of adipose tissue-derived MSC (ADSC) in order to avoid the use of xenogeneic serum. METHODS: We compared fetal bovine serum (FBS) with 10% human platelet-rich plasma (hPRP), 3% human platelet-poor plasma (hPPP) and with a cytokine cocktail composed of epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) and platelet-derived growth factor-bb (PDGFbb) added to 3% hPPP. This mixture was developed testing EGF, bFGF, granulocyte-colony stimulating factor (G-CSF), hepatocyte growth factor (HGF), insulin-like growth factor (IGF-I), PDGFbb and transforming growth factor (TGF)-beta1 added alone or in combination with hPPP. RESULTS: Our data demonstrate that the addition of EGF, bFGF and PDGFbb, in a medium supplemented with hPPP, obtainable from 150-200 mL whole autologous blood, supports ADSC expansion better than FBS, as confirmed by cumulative population doublings (cPD; 15.0 +/- 0.5 versus 9.4 +/- 2.8). The addition of human platelet-rich plasma (hPRP) further improved ADSC proliferation (cPD 20.0 +/- 1.2), but the achievement of hPRP presented a major drawback, requiring 1000-1200 mL autologous or donor whole blood. The medium supplements did not influence ADSC phenotype: they expressed CD105, CD90 and CD44 lacking hematopoietic antigens. The exposure to the proposed cocktail or to hPRP increased adipogenic and osteogenic differentiation. CONCLUSIONS: The addition of EGF, bFGF and PDGFbb to hPPP could ensure a sufficient number of ADSC for clinical applications, avoiding the use of animal serum and representing a novel approach in regenerative medicine. PMID- 21623670 TI - Oestrogen and neuroprotection. PMID- 21623671 TI - 'Prodromal diagnosis' of psychosis: an impartial commentary. PMID- 21623672 TI - Symptoms in patients with ulcerative colitis in remission are associated with visceral hypersensitivity and mast cell activity. AB - OBJECTIVE: Patients with ulcerative colitis in remission (UCR) frequently report irritable bowel syndrome (IBS)-like symptoms. Recent studies have pointed to the role of mast cells in mediating visceral hypersensitivity in IBS. We hypothesized that visceral hypersensitivity is frequently present in patients with UCR and is related to the quantity and activity of mast cells in the sigmoid mucosa. MATERIAL AND METHODS: A group of 17 controls and 19 patients with UCR were studied. Rectal compliance and perception were measured by electronic barostat. Sigmoid biopsies were taken to quantify the amount of mast cells, degranulating mast cells and mast cells in close proximity to mucosal nerve endings. RESULTS: Visceroperception significantly increased in UCR (p < 0.05) versus controls. Rectal perception correlated positively with IBS-like symptoms in UCR (r = 0.969; p < 0.05). The amount of mucosal mast cells (per 100 crypts) was significantly increased in UCR versus controls: 228 +/- 20 versus 163 +/- 18 (p < 0.05). In the UCR patients a higher percentage of mucosal mast cells was in close proximity to nerve endings (58 +/- 4 vs. 38 +/- 3% in controls; p < 0.05) or was degranulating (40 +/- 7 vs. 16 +/- 4% in controls; p < 0.05). There was a significant but weak correlation between quantity of mucosal mast cells and pain perception (r = 0.32; p < 0.05). CONCLUSION: Rectal hypersensitivity is associated with mucosal presence and activation of mast cells and with IBS-like symptoms in patients with UCR. PMID- 21623673 TI - Effects of Sandostatin LAR on gastrointestinal motility in patients with neuroendocrine tumors. AB - BACKGROUND: Diarrhea is part of the carcinoid syndrome and a significant clinical problem in neuroendocrine tumor (NET) patients. Somatostatin analog (SA) treatment usually alleviates carcinoid diarrhea, but little is known about the objective effects of SA on gastrointestinal transport. AIM: To compare gastrointestinal motility in healthy subjects and NET patients before and during SA treatment. METHODS: Twelve NET patients were studied before and during 4 weeks of SA treatment and were compared with 12 healthy controls. Radio-opaque markers were used for the assessment of total gastrointestinal transit time (GITT). Gastric and small intestinal (SI) transit patterns were described via the external tracking of a small magnetic pill ingested by the subjects. RESULTS: Compared with controls, NET patients had a significantly shorter GITT (0.7 days (0.5-1.5) vs. 1.9 days (1.0-2.3)), a shorter SI transit time (184 min (74-307) vs. 322 min (131-376)), and a faster SI velocity (2.16 cm/min (0.91-3.66) vs. 1.29 cm/min (0.76-2.60)) (all p < 0.05) but a similar gastric emptying time. SA treatment was followed by a reduction in bowel movements (five per day (3-12) vs. four per day (1-7; p < 0.02)) as well as an increase in GITT (1.4 days (0.5-2.2; p < 0.05)). Further, a trend was observed toward increased SI transit time (253 min (145-344; p = 0.08)). Gastric emptying time increased during SA treatment (19 min (4-200) vs. 179 min (5-389; p < 0.02)). Elevated chromogranin A (CgA), serotonin, and urinary 5-hydroxyindoleacetic acid (U-5HIAA) levels decreased during SA treatment. CONCLUSION: NET patients have faster than normal total GITT and SI transit times. SA treatment prolongs gastric emptying and GITT, thereby reducing the number of bowel movements. PMID- 21623674 TI - Eosinophilic gastroenteritis: clinical manifestations and morphological characteristics, a retrospective study of 42 patients. AB - OBJECTIVE: To evaluate the clinical manifestations, endoscopic, and radiological characteristics, histological features, and treatment of eosinophilic gastroenteritis in adult patients. METHODS: The clinical records of 34 patients diagnosed as eosinophilic gastroenteritis and eight patients who had abdominal symptoms and unexplained peripheral eosinophilia but no evidence of tissue eosinophilic infiltration were reviewed and analyzed. RESULTS: Thirteen patients had a history of asthma or allergic rhinitis, while 10 had an allergic history. Peripheral eosinophilia occurred in 83.3% of patients, IgE was elevated in half, and alpha2-macroglobulin was elevated in 92.8% of patients. The small intestine(82.4%)was the most common site involved. Erythema was the predominate feature seen on endoscopy. Radiographic study generally revealed thickening of the small intestinal wall. Eosinophilia generally involved the lamina propria of mucosa, but any layer of the gut could be affected, even in sites which had normal endoscopic or radiographic appearance. In all, 15.4% (6/39) of patients had Helicobacter pylori infection. Symptom remission within 1 week was found in 80% (20/25) of patients treated with steroids and in 58.8% (10/17) of patients treated with symptomatic treatment. In all, 85.7% (18/21) of patients had their eosinophil counts return to normal within 2 weeks in the steroid treatment group, but none (0/13) in the other group. CONCLUSION: Eosinophilic gastroenteritis may be more common than previously recognized. Multiple biopsies obtained from the both normal and abnormal appearing areas in the second part of the duodenum are highly recommended. Steroids are effective in relieving symptoms and improving eosinophilia. PMID- 21623675 TI - Effect of dedicated and supervised training on achieving competence in EUS-FNA of solid pancreatic lesions. AB - BACKGROUND AND AIM: The diagnostic accuracy of endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) has been suggested as a benchmark of quality performance in EUS. However, there is paucity of data on the training requirement for competency in EUS-FNA of the pancreas. KO commenced the service without prior formal training in EUS-FNA. A formally trained colleague (MN) who underwent a fellowship in the same unit was appointed to a substantive post in 2007. The aims of the study were to assess if a dedicated training program in pancreaticobiliary (PB) EUS-FNA of solid lesions: (1) produced better results at the outset of independent practice than produced at the initiation of service without formal training and (2) produced results comparable with those of an experienced endosonographer. MATERIAL AND METHODS: This is a retrospective review comparing the first 80 consecutive cases at the onset of practice of operator KO1 (2003/2004) and MN (2007/2008) as well as consecutive cases of operator KO2 (2007/2008) in the same time frame as the initial cases of operator MN. RESULTS: There was a significant difference in EUS-FNA sensitivity for pancreatic malignancy between operator KO1 (56%) and operator MN (77%) p < 0.05. There was no significant difference in test performance between operator KO2 (82%) and MN (77%) (p > 0.05). CONCLUSION: Our data show that formal training in PB EUS produces test performance at the outset of independent practice that is comparable with an experienced endosonographer, in line with the published standards for EUS-FNA of the pancreas and significantly better than that achieved without training. PMID- 21623676 TI - Emergence of entecavir-resistant mutations in nucleos(t)ide-naive Japanese patients infected with hepatitis B virus: virological breakthrough is also dependent on adherence to medication. AB - OBJECTIVE: Currently, five nucleos(t)ide analogues (NUCs) are available for the treatment of chronic hepatitis B in the world. We examined the prevalence of hepatitis B virus (HBV) DNA and alanine aminotransferase normalization in patients receiving entecavir (ETV) and the frequency of ETV-resistant mutations during an approximately 27-month use of ETV in chronic hepatitis B patients in an urban hospital in Japan. MATERIALS AND METHODS: A retrospective analysis of 81 NUC-naive chronic hepatitis B patients who received 0.5 mg of ETV daily was performed. HBV DNA was measured and sequence analysis of HBV DNA was performed in virological breakthrough patients. RESULTS: Hepatitis B e antigen (HBeAg) positive patients with HBV DNA 5.0-7.0 log IU/mL group and all HBeAg-negative patients achieved serum HBV DNA negativity by 12 months. Four patients experienced virological breakthrough during ETV therapy. Two patients had no genotypic mutations, and medical interviews revealed that they had poor adherence to ETV. CONCLUSIONS: We found that some of the HBV virological breakthroughs during ETV treatment were related to poor adherence to medication, highlighting that clinicians should pay attention to the emergence of resistant mutants as well as adherence to ETV. PMID- 21623677 TI - Comparison of non-invasive assessment to diagnose liver fibrosis in chronic hepatitis B and C patients. AB - OBJECTIVE: Chronic viral hepatitis B and C cause liver fibrosis, leading to cirrhosis. Fibrosis assessment is essential to establish prognosis and treatment indication. We compared seven non-invasive tests, separately and in combination, in chronic hepatitis patients to detect early stages of fibrosis according to the Metavir score in liver biopsy. MATERIAL AND METHODS: Galactose and methacetin breath tests (GBT and MBT), biomarkers (hyaluronic acid (HA), aspartate aminotransferase platelet ratio index (APRI), FibroTest, and Fib-4) and transient elastography (TE) were evaluated in 89 patients. Additionally, 31 healthy controls were included for evaluation of breath tests and biomarkers. RESULTS: Serum markers (HA, APRI, FibroTest, and Fib-4) and elastography significantly distinguished non-cirrhotic (F0123) from cirrhotic (F4) patients (p < 0.001, p = 0.015, p < 0.001, p = 0.005, p = 0.006, respectively). GBT, HA, APRI, FibroTest, Fib-4, and TE detected F01 from F234 (p = 0.04, p = 0.011, p = 0.009, p < 0.001, p < 0.001, and p < 0.001, respectively). A combination of different tests (TE, HA, and FibroTest) improved the performance statistically, area under the curve (AUC) = 0.87 for F234, 0.92 for F34, and 0.90 for F4. CONCLUSION: HA, APRI, FibroTest, Fib-4, and TE reliably distinguish non-cirrhotic and cirrhotic patients. Except for MBT, all tests discriminate between mild and moderate fibrosis. As single tests: FibroTest, Fib-4, and TE were the most accurate for detecting early fibrosis; combining different non-invasive tests increased the accuracy for detection of liver fibrosis to such an extent and thus might be acceptable to replace liver biopsy. PMID- 21623678 TI - Genetic structure of landraces in foxtail millet (Setaria italica (L.) P. Beauv.) revealed with transposon display and interpretation to crop evolution of foxtail millet. AB - Although the origin and domestication process of foxtail millet (Setaria italica subsp. italica (L.) P. Beauv.) has been studied by several groups, the issue is still ambiguous. It is essential to resolve this issue by studying a large number of accessions with sufficient markers covering the entire genome. Genetic structures were analyzed by transposon display (TD) using 425 accessions of foxtail millet and 12 of the wild ancestor green foxtail (Setaria italica subsp. viridis (L.) P. Beauv.). We used three recently active transposons (TSI-1, TSI-7, and TSI-10) as genome-wide markers and succeeded in demonstrating geographical structures of the foxtail millet. A neighbor-joining dendrogram based on TD grouped the foxtail millet accessions into eight major clusters, each of which consisted of accessions collected from adjacent geographical areas. Eleven out of 12 green foxtail accessions were grouped separately from the clusters of foxtail millet. These results indicated strong regional differentiations and a long history of cultivation in each region. Furthermore, we discuss the relationship between foxtail millet and green foxtail and suggest a monophyletic origin of foxtail millet domestication. PMID- 21623679 TI - Comparison of vascular stiffness in vascular dementia, Alzheimer dementia and cognitive impairment. AB - Abstract Defining the vascular component(s) of the clinical diagnosis of vascular cognitive impairment (VCI) and vascular dementia (VaD) continues to be problematic. The goal of this study was to determine whether vascular stiffness, measured by pulse wave velocity (PWV), is altered in VaD, to study the utility of PWV in differentiating VaD from Alzheimer dementia (AD) and the relationship between PWV and cognitive function. A qualitative and quantitative structured analysis of the literature was conducted until September 2010, using a search strategy based on the key words: dementia, vascular dementia, dementia of vascular origin, cognitive function and arterial stiffness or pulse wave velocity. Seventeen studies assessed large vessel vascular stiff by PWV and related it to cognitive function or dementia. Six of these studies compared PWV in 154 persons with VaD, 207 with AD and 197 controls without dementia. Mean PWV was significantly (p < 0.0001) higher in VaD compared with controls. Mean PWV was significantly (p = 0.002) higher in VaD compared with AD. Fourteen studies examined the relationship between PWV and cognitive function. The majority of studies (nine of 14) reported a significant correlation between PWV and cognitive function. Four of eight studies that evaluated the relation using univariate analysis reported a significant correlation of PWV with the Mini Mental State Exam (MMSE) or Hasegawa Dementia Scale, and the correlation with MMSE between studies showed a close agreement of correlation coefficients (0.206 to 0.27). In multivariate analysis, adjusted for a wide range of possible confounding factors, the majority or 80% (eight out of 10) studies comprising a population of 6,034 individuals found a significant inverse relationship between PWV and cognitive function. In summary, vascular stiffness is inversely related to cognitive function. Vascular stiffness is greater in VaD compared with AD, suggesting PWV may be useful in identifying VaD. PMID- 21623680 TI - Time-dependent effect of GABA on glucose-stimulated insulin secretion from isolated islets in rat. AB - OBJECTIVE: Gamma-amino butyric acid (GABA) is an inhibitory neurotransmitter found widely in the central nervous system and some peripheral tissues such as the islets of pancreas. The aim of this study was to determine the time dependency of the effect of GABA on glucose-stimulated insulin secretion in isolated islets of rats. MATERIALS AND METHODS: The collagenase digestion technique was used to isolate the islets from pancreata of male Wistar rats (200 250 g). Insulin secretion was assessed in islets exposed to different concentrations of glucose (8.3 and 16.7 mM), in the presence or absence of GABA (50 uM), for different incubation times and also pre-incubation of islets with GABA, 30 and 45 min before 8.3 mM glucose stimulation. Furthermore insulin secretion with different concentrations of glucose (8.3 and 16.7 mM), and in the presence or absence of different concentrations of baclofen, a GABA(B) agonist and of saclofen, a GABA(B) antagonist during 1 h of incubation was assessed. Insulin release was reported as mean +/- SE MUU/islet/min and p values of <0.05 were considered significant. RESULTS: GABA with concentration of 50 uM inhibited the glucose stimulated insulin secretion during a 60 min incubation period. Baclofen had no significant effect on glucose-induced insulin secretion, while saclofen (100 uM) significantly increased 16.7 mM glucose-induced insulin secretion in 60 min of incubation (p < 0.05). Pre-incubation of islets with GABA for 45 min, before glucose stimulation, increased glucose-induced insulin secretion. CONCLUSION: GABA could have both stimulatory and inhibitory effects on glucose-stimulated insulin secretion, depending on the time of exposure and glucose concentration. PMID- 21623682 TI - Levosimendan decreases intracranial pressure after hypothermic circulatory arrest in a porcine model. AB - OBJECTIVES: Hypothermic circulatory arrest (HCA) provides an optimal operating field in aortic arch surgery, but it is associated with neurological complications. Levosimendan is an inotropic agent with clinical indications for open-heart surgery. Through peripheral vasodilatation, cardiac contractility enhancement and anti-inflammatory function it has a potential to improve cerebral protection after HCA. DESIGN: Eighteen piglets were randomly assigned to a levosimendan group (n = 9) and a placebo group (n = 9) and underwent a 60-minute period of hypothermic circulatory arrest at 18 degrees C. A levosimendan or placebo infusion (0.2 MUg/kg/min) was commenced at the onset of anesthesia and continued for 24 hours. Animals were followed for one week and their neurological recovery was assessed daily. Finally the animals were electively sacrificed and their brain was harvested for histopathological examination. RESULTS: Levosimendan decreased intracranial pressure during the experiment. There were no differences between the groups in terms of hemodynamic or metabolic data, brain metabolism, neurological recovery or histopathology of the cerebral tissue. In the levosimendan group, cardiac enzymes were slightly more elevated. CONCLUSIONS: Levosimendan decreased intracranial pressure during the experiment, but in terms of cerebral metabolism, neurological recovery and histopathology of the brain tissue levosimendan did not improve brain protection in this experimental setting. PMID- 21623683 TI - Refined fructose and cancer. AB - INTRODUCTION: Caloric excess, including increased refined carbohydrate intake, is associated with higher cancer risk emphasizing the importance of improved understanding of cancer cell metabolism in tumor survival and metastasis. AREAS COVERED: This article reviews the relationship between increased dietary refined sugar and cancer risk, with specific emphasis on the monosaccharide fructose. Cancer cell metabolism is reviewed, and the potential mechanisms by which dietary sugars contribute to the tumor microenvironment are discussed. Recent observations indicate that cancer cells readily utilize fructose to support proliferation and preferentially use fructose for nucleic acid synthesis. This review discusses the potential role of how dietary fructose can promote cancer growth by a variety of mechanisms, including altered cellular metabolism, increased reactive oxygen species, DNA damage and inflammation. Preliminary insights into potential therapeutic strategies by which fructose-mediated cancer effects may be abrogated are presented. EXPERT OPINION: Other sugars (particularly fructose, given its abundance in the modern diet) must be considered with reference to cancer cell metabolism. Cancer cells utilize similar sugars in distinct ways, which may present important new therapeutic avenues of targeting cancer. PMID- 21623684 TI - Wnt-signaling pathways in progressive renal fibrosis. AB - INTRODUCTION: The prevention and potential reversal of interstitial fibrosis is a central strategy for the treatment of progressive renal disease. This strategy requires a better understanding of the underlying pathophysiologic processes involved in progressive renal fibrosis. AREAS COVERED: The developmental processes in which Wnt (combination of 'wingless' and 'INT')/frizzled signaling is involved is discussed in this review, including cell fate determination, cell polarity, tissue patterning and control of cell proliferation. These pathways are also active in the adult where they play key roles in the maintenance of tissue homeostasis, wound repair and chronic tissue damage. EXPERT OPINION: Wnt biology helps to control cell polarity, moderates cell proliferation and underlies other processes linked to renal homeostasis. Reactivation and dysregulation of the Wnt pathways underlie chronic fibrosis and progressive renal failure. Wnt signaling is, however, context-dependent: the pathways are complex and undergo many levels of cross-talk with other regulatory systems and regulatory pathways. On one hand, this may help to explain the positive effects of Wnt-signaling blockades seen in some animal models of chronic renal damage and, on the other, this suggests that it may be difficult to predict how modifications of the Wnt pathway may influence a process. PMID- 21623681 TI - Fitness and fatness are independently associated with markers of insulin resistance in European adolescents; the HELENA study. AB - OBJECTIVE: To examine the independent association of total and central body fat and cardiorespiratory fitness with markers of insulin resistance after controlling for several potential confounders in European adolescents participating in the HELENA-CSS (Healthy Lifestyle in Europe by Nutrition in Adolescence Cross-Sectional) study. METHODS: We conducted a cross sectional study (the HELENA-CSS) which comprised 1053 (12.5-17.5 years) adolescents from 10 European cities. Weight, height, waist circumference and skinfold thickness were measured, and body mass index (BMI) was calculated. Cardiorespiratory fitness was measured by the 20-m shuttle run test. Markers of insulin resistance were fasting insulin and glucose, and homeostasis model assessment (HOMA). RESULTS: HOMA and insulin were positively associated with BMI, skinfolds and waist circumference after controlling for center, age, pubertal status and cardiorespiratory fitness (all P ? 0.01). HOMA and insulin were negatively associated with cardiorespiratory fitness in adolescents with moderate to high levels of total and central body fat (all P ? 0.01). CONCLUSIONS: HOMA and insulin were associated with total and central body fat in European adolescents. Moreover, cardiorespiratory fitness explained a part of the HOMA and insulin variance in those adolescents with moderate to high levels of total and central body fat, and also, to some extent, in those with low to middle fat mass. PMID- 21623685 TI - A systematic review of the efficacy and safety outcomes of anti-VEGF agents used for treating neovascular age-related macular degeneration: comparison of ranibizumab and bevacizumab. AB - OBJECTIVE: To systematically review ocular and systemic events in treatment of wet age-related macular degeneration (AMD) with anti-vascular endothelial growth factor antibodies, ranibizumab and bevacizumab, and to provide a detailed perspective of their differences on clinical use, efficacy and safety. RESEARCH DESIGN AND METHODS: This review was based on a 2010 PubMed literature search performed using two separate terms: 'lucentis' OR 'ranibizumab' AND 'age-related macular degeneration' OR 'AMD' or 'avastin' OR 'bevacizumab' AND 'age-related macular degeneration' OR 'AMD'. A clinical diagnosis of wet AMD was defined by the authors of the trial reports. Clinical studies that met Level I or Level II evidence criteria were considered for review. FINDINGS: Eight large, randomized, controlled trials of ranibizumab (Level I) included 1,485 patients (range 162 716) and four open-label studies of ranibizumab (Level II) included 4,484 patients (range 32-4,300). Six studies (one Level I, five Level II) of bevacizumab included 424 patients (range 28-165). All demonstrated improvements in visual acuity. Only one study (Level II) compared the efficacy of ranibizumab and bevacizumab. Adverse ocular and systemic safety events occurring during the study were prospectively recorded for ranibizumab, irrespective of their suspected relationship to study treatments. Only three of six bevacizumab studies reported details of adverse ocular or systemic events. There was extensive Level I and Level II evidence to support both the efficacy and safety of ranibizumab in wet AMD. Data suggest that bevacizumab provides efficacy in wet AMD, but the safety profile of intravitreal bevacizumab remains to be established. CONCLUSION: In contrast to ranibizumab, current safety data for bevacizumab are incomplete and not yet robust. If the medical community remains committed to using intravitreal bevacizumab, it is critical to establish that it has an acceptable safety profile, supported by evidence-based medicine. Considerable further research is warranted to achieve this. PMID- 21623686 TI - New targets, new drugs for metastatic bone pain: a new philosophy. AB - Bone pain is a common symptom in bone metastases. The therapies that are currently available include nonsteroidal anti-inflammatory drugs (NSAIDs), opioids, steroids and gabapentin which have been demonstrated to improve neuropathic pain. In addition, preclinical studies indicate that agents such as transient receptor potential vanilloid 1 antagonists and cannabinoid 2 receptor agonist could be considered as adjuncts in ameliorating opioid side effects. New drugs are in the clinical phase of development, among which the most promising molecules seem to be anti-nerve growth factor (NGF) antibodies. Anti-NGF antibody therapy may be particularly effective in blocking bone cancer pain because NGF appears to be integrally involved in the upregulation, sensitization and disinhibition of multiple neurotransmitters, ion channels and receptors in the primary afferent nerve. The best way to treat bone metastases pain is to improve the control of skeletal disease burden. Recently, denosumab, a noncytotoxic IgG2 monoclonal antibody with high affinity for human RANKL, has been demonstrated to significantly prevent clinically relevant increase in pain compared with zoledronic acid across the tumor types. Based on these data, it has been suggested that denosumab has the potential to become a new standard of treatment in bone metastases management. PMID- 21623687 TI - Effect of NADPH-oxidase inhibitors in the experimental model of zymosan-induced shock in mice. AB - The aim of this study was to investigate the effects of NADPH-oxidase inhibitors, in a mouse model of zymosan. Zymosan-induced shock was induced in mice by administration of zymosan (500 mg/kg, i.p.). The pharmacological treatment was the administration of apocynin (5 mg/kg 10% DMSO i.p.) and diphenylene iodonium chloride (DPI) (1 mg/kg i.v.) 1 h and 6 h after zymosan administration. MOF and systemic inflammation in mice was assessed 18 h after administration of zymosan. NADPH-oxidase inhibitors caused a significant reduction of the (1) peritoneal exudate formation, (2) neutrophil infiltration, (3) multiple organ dysfunction syndrome, (4) nitrotyrosine, (5) poly (ADP-ribose) (PAR), (6) cytokine formation, (7) adhesion molecule expression, (8) nuclear factor (NF-kappaB) expression and (9) apoptosis induced by zymosan. Moreover, NADPH-oxidase inhibitors treatment significantly reduced the systemic toxicity, the loss in body weight and the mortality caused by zymosan. This study has shown that NADPH-oxidase inhibitors attenuate the degree of zymosan-induced non-septic shock in mice. PMID- 21623688 TI - Enhancing effect of Labrasol on the intestinal absorption of ganciclovir in rats. AB - Ganciclovir (GCV), like other nucleoside analogs such as trifluridine and acyclovir (ACV), is hydrophilic, poorly permeable across membranes and orally low bioavailable. In the present studies, Labrasol was evaluated for improving intestinal absorption of GCV through in vitro and in vivo experiments. The effect of Labrasol on absorption of GCV in rat small intestine was investigated using an in situ single-pass perfusion technique. The apparent absorptive clearance (PeA) of GCV with Labrasol in the duodenum, jejunum and ileum was 1.01, 1.28, and 1.49 mL/min/cm (n = 6), respectively, and significant regional differences of GCV absorption among the three segments were observed p jejunum, p duodenum (p > 0.05). The effects of EDTA, verapamil on the permeability of GCV were conducted. The permeability of GCV was increased by EDTA, verapamil, respectively. The results indicated that paracellular absorption and efflux played important roles in GCV absorption. In vivo absorption GCV in rats was conducted. When GCV at 1 mg/kg dose was administered with Labrasol (10%, v/v), the mean AUC of was determined as 14.45 +/- 3.88 MUg*h/mL, compared to 8.05 +/- 1.52 ug*h/mL without Labrasol. Based on the results, we could conclude that the absorption of GCV through GI lumen would be enhanced by Labrasol. The effect of Labrasol maybe ascribed to both (i) inhibit efflux of GCV from the enterocytes to the GI lumen; and (ii) enhance GCV absorption from the GI lumen through paracellular pathway. PMID- 21623689 TI - Aldo-keto reductase 1C15 as a quinone reductase in rat endothelial cell: its involvement in redox cycling of 9,10-phenanthrenequinone. AB - 9,10-Phenanthrenequinone (9,10-PQ), a redox-active quinone in diesel exhausts, triggers cellular apoptosis via reactive oxygen species (ROS) generation in its redox cycling. This study found that induction of CCAAT/enhancer-binding protein homologous protein (CHOP), a pro-apoptotic factor derived from endoplasmic reticulum stress, participates in the mechanism of rat endothelial cell damage. The 9,10-PQ-mediated CHOP induction was strengthened by a proteasome inhibitor (MG132) and the MG132-induced cell sensitization to the 9,10-PQ toxicity was abolished by a ROS inhibitor, suggesting that ROS generation and consequent proteasomal dysfunction are responsible for the CHOP up-regulation caused by 9,10 PQ. Aldo-keto reductase (AKR) 1C15 expressed in rat endothelial cells reduced 9,10-PQ into 9,10-dihydroxyphenanthrene concomitantly with superoxide anion formation, implying its participation in evoking the 9,10-PQ-redox cycling. The 9,10-PQ-induced damage was augmented by AKR1C15 over-expression. 9,10-PQ also provoked the AKR1C15 up-regulation, which sensitized against the quinone toxicity. These results suggest the presence of a negative feedback loop exacerbating the quinone toxicity in rat endothelial cells. PMID- 21623690 TI - Inhaler technique maintenance: gaining an understanding from the patient's perspective. AB - BACKGROUND: The aim of this study was to determine the patient-, education-, and device-related factors that predict inhaler technique maintenance. METHODS: Thirty-one community pharmacists were trained to deliver inhaler technique education to people with asthma. Pharmacists evaluated (based on published checklists), and where appropriate, delivered inhaler technique education to patients (participants) in the community pharmacy at baseline (Visit 1) and 1 month later (Visit 2). Data were collected on participant demographics, asthma history, current asthma control, history of inhaler technique education, and a range of psychosocial aspects of disease management (including adherence to medication, motivation for correct technique, beliefs regarding the importance of maintaining correct technique, and necessity and concern beliefs regarding preventer therapy). Stepwise backward logistic regression was used to identify the predictors of inhaler technique maintenance at 1 month. RESULTS: In total 145 and 127 participants completed Visits 1 and 2, respectively. At baseline, 17% of patients (n = 24) demonstrated correct technique (score 11/11) which increased to 100% (n = 139) after remedial education by pharmacists. At follow-up, 61% (n = 77) of patients demonstrated correct technique. The predictors of inhaler technique maintenance based on the logistic regression model (X(2) (3, N = 125) = 16.22, p = .001) were use of a dry powder inhaler over a pressurized metered-dose inhaler (OR 2.6), having better asthma control at baseline (OR 2.3), and being more motivated to practice correct inhaler technique (OR 1.2). CONCLUSION: Contrary to what is typically recommended in previous research, correct inhaler technique maintenance may involve more than repetition of instructions. This study found that past technique education factors had no bearing on technique maintenance, whereas patient psychosocial factors (motivation) did. PMID- 21623691 TI - Cladribine plus rituximab is an effective therapy for newly diagnosed mantle cell lymphoma. AB - Mantle cell lymphoma (MCL) is a non-Hodgkin lymphoma that is incurable with standard chemotherapy. There is no consensus on the best initial therapy, especially for elderly patients, who are not candidates for aggressive treatment approaches. Current National Comprehensive Cancer Network (NCCN) treatment guidelines include rituximab (R) plus cladribine for the initial treatment of MCL. However, few data are available to substantiate this recommendation. Therefore, to further define the role of R-cladribine for the initial treatment of MCL, we performed a retrospective chart review of 31 patients with MCL (median age, 67) treated with R-cladribine. The majority of responding patients also received R maintenance. The overall response rate was 87%, with 61% of patients achieving a complete remission (CR/CRu). The estimated median follow-up was 32.5 months, median PFS was 37.5 months, and median OS was 85.2 months. One of 19 (5.3%) subjects in CR/CRu relapsed (median follow-up of 23 months). CR/CRu was associated with improved survival (p < 0.0001), while a high mantle cell international prognostic index (MIPI) was associated with worse survival (p = 0.05). There was one toxic death (neutropenic pseudomonal sepsis) related to treatment. R-cladribine is an effective therapy for previously untreated MCL, and these results validate the use of R-cladribine for the initial treatment of MCL. PMID- 21623692 TI - Prognostic impact of MUM1 expression by immunohistochemistry on primary mediastinal large B-cell lymphoma. AB - Diffuse large B-cell lymphoma can be classified into two prognostically distinct subgroups with germinal center B-cell-like (CG) and activated B-cell-like (post CG) characteristics, based on CD10, BCL-6, and MUM1 expression. We performed a retrospective analysis of the clinical variables of 37 patients with primary mediastinal large B-cell lymphoma and the expression of BCL-6 and MUM1 in 22 patients with available tissue. The median age was 30 years, and 70% of the patients were female. BCL-6 and MUM1 were expressed in 64% and 45% of cases, respectively. Five-year overall survival (OS) and disease-free survival (DFS) were 47% and 81%, respectively. In univariate analysis, complete response (p = 0.0001), radiation therapy (p = 0.01), International Prognostic Index (p = 0.001), and MUM1 expression (p = 0.002) correlated with OS. For this group of patients with homogeneous clinical characteristics, response to initial chemotherapy and MUM1 expression were associated with prognosis. PMID- 21623693 TI - Response to "Theoretical aspects of autism: causes--a review" by Ratajczak, HV (Journal of Immunotoxicology 8:68-79, 2011). PMID- 21623694 TI - Hemoglobin-albumin crosslinked copolymers: reduced prooxidant reactivity. AB - We have previously reported that derivatization of hemoglobin with periodate modified sugar derivatives such as oxidized adenosine triphosphate (oATP) leads to an increase in prooxidant reactivity at the heme. Here, we report that copolymerization of hemoglobin with serum albumin alleviates this problem completely, to the extent where the copolymer even has a slightly lower autooxidation rate compared to native hemoglobin. A similar, although not as potent, effect is obtained when hemoglobin is derivatized with oATP in the presence of small-molecule antioxidants instead of albumin. PMID- 21623695 TI - Packing density of the PEG-shell in PEG-albumins: PEGylation induced viscosity and COP are inverse correlate of packing density. AB - PEG-Alb represents a new class of low viscogenic plasma expanders that achieve super perfusion in vivo by mimicking the vasodilatory influence of high viscogenic plasma expanders. PEGylation-engineered structure of PEG albumin can be envisaged as a deformable molecular domain around the rigid central protein core. The correlation between the structure of PEG-shell in terms of packing of the PEG inside the PEG shell and PEGylation induced plasma expander (PE)-like properties of albumin has been investigated as a function of the number and length of the PEG-chain. The increase in molecular radius of albumin on PEGylation is non-linear as a function of the number of PEG chains conjugated. The packing density of PEG within the PEG-shell is an inverse correlate of PEG chain size; i.e. the shorter chains pack more compactly than the longer ones. The PEGylation induced increase in the viscosity and COP of albumin is an exponential correlation of the number of ethylene oxide units (-CH(2)-CH(2)-O-) conjugated and is also a function of the PEG-chain length. At equivalence of PEG mass conjugated, the viscosity and COP of PEG-albumin adducts correlate inversely with packing density of PEG. All PEGylated albumins are not equivalent on the basis of total PEG mass conjugated. Accordingly, the structure of PEG albumin and its solution properties can be engineered to optimize a given total PEG mass for the application of PEG albumin as a resuscitation fluid. The extension arms minimize the influence of PEG shell on the structure of the protein core. We speculate that EAF-PEGylation is a preferable platform for PEGylation of protein therapeutics and is expected to generate products with better therapeutic efficacy. PMID- 21623696 TI - Polymerized human placenta hemoglobin (PolyPHb) attenuates myocardial infarction injury in rats. AB - OBJECTIVES: To investigate the cardioprotective effect of polymerized human placenta hemoglobin (PolyPHb) for acute myocardial ischemia rat heart. METHODS: Myocardial infarcts (MI) model was set up in SD rats by permanent ligation of the left anterior descending coronary artery (LDA). The rats were divided randomly into 3 groups with each group of 20 rats: (A) the control group with the administration of Ringer's lactated solution at a dose of 2.5ml/kg); (B) PolyPHb group, PolyPHb solution at a dose of 0.16g Hb/kg and (C), PolyPHb+CAT+SOD group, PolyPHb solution at a dose of 0.16g Hb/kg and catalase (CAT) solution and superoxide dismutase (SOD) solution at a dose of 1681 U/kg and 528000 U/kg, respectively. Each rat received treatments via caudal vein, once a day for 7 days. Qualitative evaluations were made based on the reading of cardiac troponin T (cTnT), the myocardial infarction size (MIS) derived from the staining of myocardium tissue, and the pathological changes in infarct area. The ischemia changes of cardiomyocytes were determined by haematoxylin - basic fuchsin - picric acid (HBFP) staining and the apoptosis of cardiomyocytes were evaluated by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay (TUNEL). RESULTS: Compared to the control group, PolyPHb greatly decreased the cTnT (p < 0.05), MIS (p < 0.05), and the size of myocardial ischemia (p < 0.05). PolyPHb + CAT + SOD decreased the ?ST change (P < 0.05), cTnT (P < 0.01), MIS (p < 0.05), the pathological scores (p < 0.01), the size of myocardial ischemia (p < 0.01), and the apoptosis level (p < 0.01). CONCLUSION: Our study demonstrated that adding PolyPHb improves cardiac functional recovery and reduces myocardial infarction of rat heart. PMID- 21623697 TI - Tetrabromobisphenol A decreases cell-surface proteins involved in human natural killer (NK) cell-dependent target cell lysis. AB - Human natural killer (NK) lymphocytes are able to destroy tumor cells and virally infected cells. Interference with their function can leave an individual with increased susceptibility to cancer development and/or viral infection. We have shown that the tumor-destroying (lytic) function of NK cells can be dramatically decreased by exposure to the environmental contaminant tetrabromobisphenol A (TBBPA). TBBPA is a flame retardant used in a variety of materials including circuit boards, carpeting, and upholstery and has been found in human blood samples. TBBPA interferes with NK cell lytic function, in part, by decreasing the ability of NK cells to bind to target cells. This study examines the effects of exposures to concentrations of TBBPA (i.e., that were able to decrease the binding capacity of NK cells) on the expression of cell-surface proteins (CD2, CD11a, CD16, CD18, and CD56) that are needed for NK cells to bind target cells. NK cells were exposed to TBBPA for 24 h, 48 h, and 6 days or for 1 h followed by 24 h, 48 h, and 6 days in TBBPA-free media. Twenty-four-hour exposures to 5 uM TBBPA caused decreases in four of the cell-surface proteins examined. CD16 was decreased by >35%. The decreases in cell-surface proteins after a 48-h exposure were similar to those seen after 24 h. The results indicate that TBBPA exposures that decrease the binding function of human NK cells do so by decreasing the expression of cell-surface proteins needed for attachment of NK cells to targets cells. PMID- 21623698 TI - Clinical response to statins: mechanism(s) of variable activity and adverse effects. AB - Statins represent a major advance in the treatment of hypercholesterolemia, a significant risk factor for atherosclerosis. There is, however, notable interindividual variation in the cholesterolemic response to statins, and the origin of this variability is poorly understood; pharmacogenetics has attempted to determine the role of genetic factors. Myopathy, further, has been reported in a considerable percentage of patients, but the mechanisms underlying muscle injury have yet to be fully characterized. Most statins are the substrates of several cytochrome P450s (CYP). CYP polymorphisms may be responsible for variations in hypolipidemic activity; inhibitors of CYPs, e.g. of CYP3A4, can significantly raise plasma concentrations of several statins, but consequences in terms of clinical efficacy are not uniform. Pravastatin and rosuvastatin are not susceptible to CYP inhibition but are substrates of the organic anion transporting polypeptide (OATP) 1B1, encoded by the SLCO1B1 gene. Essentially all statins are, in fact, substrates of membrane transporters: SLCO1B1 polymorphisms can decrease the liver uptake, as well as the therapeutic potential of these agents, and may be linked to their muscular side-effects. A better understanding of the mechanisms of statin handling will help to minimize adverse effects and interactions, as well as to improve their lipid-lowering efficiency. PMID- 21623699 TI - From circulating biomarkers to genomics and imaging in the prediction of cardiovascular events in the general population. AB - Cardiovascular disease (CVD) is the leading cause of death and disability worldwide. In the last decades numerous markers have been considered and investigated for the prediction of CV events, but only a few of them resulted in improved global risk assessment beyond traditional risk factors when incorporated into coronary evaluation scores. Recent genetic studies have pointed out a few but consistent loci or genes which are independently associated with CV risk. The idea is fascinating that these genetic markers could lead to improved individual CV risk assessment and tailored pharmacological interventions. In this brief review we will not make a systematic review of all non-genetic and genetic markers of CV risk but we will try to make a brief overview of the most interesting ones with the aim to underline potential 'pros' and 'cons' of their implementation in clinical practice. PMID- 21623700 TI - Therapeutic options in treatment-resistant depression. AB - The phenomenon of treatment-resistant depression (TRD), described as the occurrence of an inadequate response after an adequate treatment with antidepressant agents (in terms of dose, duration, and adherence), is very common in clinical practice. It has been broadly defined in the context of unipolar major depression, but alternative definitions for bipolar depression have also been suggested. In both cases, there is a remarkable lack of consensus amongst professionals concerning its operative definition. A relatively wide variety of treatment options for unipolar TRD are available, whilst the evidence is very scanty for bipolar TRD. TRD is associated to poor clinical, functional, and social outcomes. Several novel therapeutic options are currently being investigated as promising alternatives, targeting the neurotransmitter system outside of the standard monoamine hypothesis. Augmentation or combination with lithium or atypical antipsychotics appears as a valid option for both conditions, and the same occurs with electroconvulsive therapy. Other non-pharmacological strategies such as deep brain stimulation may be promising alternatives for the future. The use of cognitive behaviour therapy is recommended for unipolar TRD, but there is no evidence supporting its use in bipolar TRD. PMID- 21623701 TI - Cysteine effects on the pharmacokinetics of etoposide in protein-calorie malnutrition rats: increased gastrointestinal absorption by cysteine. AB - Protein-calorie malnutrition (PCM) occurs frequently in advanced cancer patients and has a profound impact on the toxicity of many drugs. Thus, the pharmacokinetics of etoposide were evaluated in control, control with cysteine (CC), PCM, and PCM with cysteine (PCMC) rats. Etoposide was administered intravenously (2 mg/kg) or orally (10 mg/kg). Changes in hepatic and intestinal cytochrome P450s (CYPs) and effects of cysteine on intestinal P-glycoprotein (P gp)-mediated efflux were also measured. In PCM rats, the CL(NR) (AUC(0-infinity)) of intravenous etoposide was significantly slower (greater) than that in controls, because of the significant decrease in the hepatic CYP3A subfamily and P-gp. In PCMC rats, the slowed CL(NR) of etoposide in PCM rats was restored to the control level by cysteine treatment. PCMC rats showed a significantly greater AUC(0-6 h) of oral etoposide than PCM rats, primarily because of the increased gastrointestinal absorption of etoposide as a result of the inhibition of intestinal P-gp by cysteine. The gastrointestinal absorption of an oral anticancer drug, which is a substrate of P-gp, may be improved by co administration of cysteine in advanced cancer patients if the present rat data can be extrapolated to patients. PMID- 21623702 TI - Enantioselective carbonyl reduction of eperisone in human liver microsomes. AB - Eperisone, 4-ethyl-2-methyl-3-piperidinopropiophenone, is a centrally acting muscle relaxant widely used to relieve muscle stiffness and back pain. In this study, enantioselectivity for carbonyl reduction of eperisone was investigated in human liver microsomes, and the enzymes involved in the carbonyl reduction were characterised. Carbonyl reduction of eperisone predominantly occurred in microsomal fractions and 11beta-hydroxysteroid dehydrogenase type 1(11beta-HSD 1) played a major role in this reaction as judged by selective inhibition of the activity by BVT-14225 and KR-66344. The kinetic study with (+)-S- and (-)-R eperisone showed that the formation of the carbonyl reduced metabolite (M5) from the (-)-R-isomer was more efficient than that from the (-)-S-isomer. As eperisone is a racemic compound with one chiral centre, the carbonyl reduced metabolite of eperisone (M5) may have four possible diastereoisomeric structures. Chiral separation of incubation mixtures of racemic eperisone with human liver microsome revealed that (1S, 2S)-M5 and (1R, 2R)-M5 were generated specifically from (+)-S- and (-)-R-eperisone, respectively. Selective formation of anti-diastereomers was further confirmed by incubation of individual enantiomer with microsomes. Carbonyl reduction of eperisone by microsomal 11beta-HSD 1 may significantly contribute to the metabolic disposition of eperisone in human and (-)-R-isomer is preferentially reduced by this enzyme. PMID- 21623703 TI - Fetal gene therapy: recent advances and current challenges. AB - INTRODUCTION: Fetal gene therapy (FGT) can potentially be applied to perinatally lethal monogenic diseases for rescuing clinically severe phenotypes, increasing the probability of intact neurological and other key functions at birth, or inducing immune tolerance to a transgenic protein to facilitate readministration of the vector/protein postnatally. As the field is still at an experimental stage, there are several important considerations regarding the practicality and the ethics of FGT. AREAS COVERED: Here, through a review of FGT studies, the authors discuss the role and applications of FGT, the progress made with animal models that simulate human development, possible adverse effects in the recipient fetus and the mother and factors that affect clinical translation. EXPERT OPINION: Although there are valid safety and ethical concerns, the authors argue that there may soon be enough convincing evidence from non-human primate models to take the next step towards clinical trials in the near future. PMID- 21623704 TI - The potential of amniotic fluid stem cells for cellular therapy and tissue engineering. AB - INTRODUCTION: Foetal cells present in amniotic fluid (AF) have been used for many years to perform prenatal genetic screening. Recent reports suggested that these cells might have additional benefits. AF contains, in addition to committed and differentiated cells, a subpopulation with stem cell characteristics. AF-derived stem cells (AFS) have functions found in mesenchymal stem cells, but in addition, exhibit a potent expansion capacity and plasticity. AFS are able to undergo multi lineage differentiation and produce progeny indicative of all three germ layers. AREAS COVERED: The experimental approaches available to isolate AFS and their potential for tissue engineering, the repair of organs through cell replacement and tissue regeneration. EXPERT OPINION: The deployment of AFS for tissue regeneration offers advantages over the use of embryonic or adult stem cells: i) AF represents a convenient and non-contested source for obtaining stem cells; ii) their derivation is relatively simple and rapid; iii) no feeder layers are required for their cultivation; iv) they display no spontaneous differentiation in culture; and v) their stem cell phenotype is not affected by long-term storage. The application of AFS for tissue replacement therapies in vivo is at a very early stage, but existing studies indicate great potential for clinical use. PMID- 21623705 TI - Chitosan-reduced gold nanoparticles: a novel carrier for the preparation of spray dried liposomes for topical delivery. AB - Exposure of skin to various chemical and physical agents results in excessive stress to the outermost cell layer of the skin, causing different degenerative effects that can be minimized by using antioxidant formulations. The major challenge, in this regard, is to develop a formulation, which can prevent photodegradation of the actives, thus allowing a significant amount to be deposited at the site. In recent decades, liposomal formulations have been extensively employed to overcome the barrier properties of the skin and photodegradation of actives. In the present study, chitosan-reduced gold nanoparticles were investigated for its potential as a carrier to prepare liposomes by a spray-drying method. Liposomes so obtained were characterized for phospholipid recovery, diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy, particle size, zeta potential, encapsulation efficiency, and deposition of drug and gold nanoparticles in the rat skin. Further, a liposomal gel formulation was prepared using Carbopol(r) 980 NF (Noveon Systems, Kochi, India) and evaluated for drug deposition in the skin. Antioxidant activity of vitamin C encapsulated in gold liposomes was determined on a human leukemia (HL 60) cell line. The use of gold nanoparticles as a carrier showed improved phospholipid recovery and thus overcomes the liposome scalability problem. DRIFT spectra confirmed the presence of phospholipid in the formulation. Liposomal gel showed improved drug deposition, as compared to control and marketed preparations. A more interesting contribution of the chitosan-reduced gold nanoparticles was an enhanced antioxidant activity seen in case of the vitamin C loaded gold liposomal formulation. Liposomal formulation was found to be stable for 3 months at 30 degrees C and 65% relative humidity. PMID- 21623706 TI - Anticomplement activity of isolated compounds from Artemisia montana. AB - The study evaluated the anticomplement activity from isolated compounds from Artemisia montana (Nakai) Pamp. from South Korea on the classical pathway. In a previous work, A. montana (Nakai) Pamp. chloroform extracts showed inhibitory activity against complement system. The chromatographic separation of a chloroform chloride extract of A. montana (Nakai) Pamp. led to the isolation of four compounds. Their structures were characterized to be ezoartemin, yamayomoginin, ezomontanin and 11,13-dihydroezomontanin by spectroscopic data. This is the first report of anticomplement activity of isolated compounds from A. montana (Nakai) Pamp. PMID- 21623707 TI - Interplay of drug metabolizing CYP450 enzymes and ABC transporters in the blood brain barrier. AB - The recent identification of drug-metabolizing enzymes cytochrome P450 (CYP) in the human blood-brain barrier (BBB) raises the question of whether these enzymes act in concert with ATP-binding cassette (ABC) transporters to limit the brain distributions of drugs. We recently demonstrated several CYP genes in freshly isolated human brain microvessels; the main isoforms expressed were CYP1B1 and CYP2U1. Many studies using different experimental approaches have revealed that P glycoprotein (P-gp, ABCB1), breast cancer resistance protein (BCRP, ABCG2) and the multidrug resistance-associated protein 4 (MRP4, ABCC4) are the main ABC transporters in the human BBB. The first part of this review covers recent studies on the expression, regulation and function of CYP450 and ABC transporters in the rodent and human BBBs. The second part focuses on the possible interplay between some CYPs and certain ABC transporters at the BBB, which makes it a determining element of brain drug concentrations and thus of the effects of centrally acting drugs. PMID- 21623708 TI - Bleeding, vertebral fractures and vascular calcifications in patients treated with warfarin: hope for lower risks with alternative therapies. AB - Anticoagulant therapy in patients with atrial fibrillation requires careful evaluation because its benefits i.e. prevention of thromboembolism, must be greater than the risk of bleeding. Patients at higher risk of thrombosis are evaluated through specific scores, such as the CHA(2)DS(2)VASc, coupled with scoring systems for assessing bleeding risks, such as the HAS-BLED score. In addition to bleeding, other risks have been associated with the use of warfarin, including an increased susceptibility to vascular calcifications and fractures caused by a reduction in the levels of vitamin K dependent carboxylated enzymes, matrix Gla-protein (MGP) and bone Gla-protein or osteocalcin (BGP). In fact, while on one side warfarin is used to prevent embolism, on the other hand acting as a vitamin K antagonist it blocks the inhibitory effect of MGP on vascular calcification. Similarly, patients treated with warfarin carry a greater risk of developing osteoporosis and fractures, due to reduced BGP activity. Recently, a new generation of anticoagulant drugs has been developed, such as dabigatran, a direct thrombin inhibitor, and rivaroxaban, a direct factor-Xa inhibitor. They offer an interesting alternative to warfarin, because they do not require frequent blood tests for monitoring while offering similar results in terms of efficacy. Lacking the inhibitory effect on the vitamin K cycle, the consequent side effects can be avoided. If, compared to warfarin treated patients, a lower incidence of vascular calcifications and fractures will be demonstrated, the advantages over warfarin may be even greater, leading to further benefits in terms of morbidity and mortality. PMID- 21623713 TI - [Data of coronary angiography and rheological properties of erythrocytes in non ST-elevation acute coronary syndrome]. AB - We assessed character and extent of coronary vessels involvement in 27 patients with non ST-elevation acute coronary syndrome (NSTEACS) and compared angiography data with rheological properties of erythrocytes of patients and 30 practically healthy men. Stenoses of main branches of coronary vessels were found almost in 3/4 of cases. In 4 patients without stenoses in main branches of coronary arteries we revealed lowering of blood flow velocity related to small vessel disease (syndrome X). In 2 cases there were no changes in coronary circulation. Extent of vessel involvement was the least in patients with non Q-wave myocardial infarction as initial manifestation of ischemic heart disease and the greatest in patients with unstable angina after myocardial infarction. Patients with NSTEACS had elevated aggregation capacity and lowered deformability of erythrocytes. Degree of these changes did not depend on clinical peculiarities of development of the disease, extent of obstructions, character of vessels involvement. Positive correlation between was found content of cholesterol and degree of elevation of rate of erythrocyte aggregation. PMID- 21623714 TI - [Predictors of stent thrombosis in patients with ST-elevation acute coronary syndrome subjected to primary coronary artery intervention]. AB - Basing on complex evaluation of clinical, angiographic, and laboratory data we revealed predictors of stent thrombosis in patients with ST-elevation acute coronary syndrome (ACS) subjected to percutaneous coronary interventions (PCI). Among studied sample of patients (n=124) we distinguished a group of patients with proven stent thrombosis at various stages of follow-up (n=22, group 1) and a group of patients with favorable outcome (n=102, group 2). Unifactorial analysis revealed unfavorable prognostic influence (p=0.025) of hemodynamically significant stenosis of proximal segment of anterior distending artery, disturbances of rhythm and conduction (p=0.0002), as well as degree of heart failure (HF) according to Killip on day 1 of development of symptoms of ACS, and also of high selectin P level on day 10 (p=0.031). With the aim of assessment of prognostic significance of revealed factors we conducted stepwise discriminant analysis according to results of which most significant parameter affecting development of stent thrombosis in patients with ST-elevation ACS appeared to be high Killip class of HF (p<0.0003), to a lesser degree - elevated level of sP selectin on day 10 of observation (p=0.005). Parameter for which prognostic significance was not revealed as a result of multifactorial analysis was lesion in proximal segment of anterior descending artery (p=0.496). Probability of correct classification of the model was 88.7% at p<0.0001. PMID- 21623715 TI - [Clinical- functional assessment of organoprotective efficacy of enalapril and telmisartan in patients with arterial hypertension]. AB - Effect of enalapril and telmisartan on hemodynamic indices, structural-functional parameters of the left ventricle, and vasomotor function of arterial endothelium was studied in 49 patients (mean age 48.3+/-3.2 years) with 1-2 degree arterial hypertension (AH) and abnormal relaxation type of left ventricular diastolic dysfunction. Enalapril was given to 24 and telmisartan to 25 patients. Examination included 24-hour arterial pressure monitoring, echocardiography, duplex scanning of the left brachial artery. At the background of therapy with enalapril target levels of systolic and diastolic arterial pressure (AP) were achieved in 79.2 and 70.85% of patients. This was associated with 54.2% reduction of number of non-dippers and night peakers. Therapy with telmisartan (30 weeks) besides effective AP control provided normalization of 24-hour AP profile in 90% of patients with pathological circadian rhythm. This was accompanied by improvement of vasomotor endothelial function of arteries, positive dynamics of structural-functional parameters of the left heart chambers, improvement of indices of left ventricular diastolic function. PMID- 21623716 TI - [Selection of rational combinations of indapamide with various of calcium antagonists in patients with arterial hypertension]. AB - We studied 89 patients with II degree arterial hypertension (age 43-67 years). Before and in 24 weeks after therapy we carried out 24-hour monitoring of arterial pressure and Doppler echocardiography. Patients were divided into 2 groups. Combination therapy with indapamide and verapamil retard, indapamide and amlodipine were prescribed to patients of group 1 and 2, respectively. At the background of therapy circadian rhythm was normalized in 82.8 and 76.9% of patients in groups 1 and 2, respectively. In group1 therapy was especially effective in patients with type 1 left ventricular diastolic dysfunction (LVDD), 24-hour nondipper profile, and hyperkinetic type of circulation. In group 2 pronounced changes of parameters were seen in II-III type of LVDD and hypokinetic type of circulation. PMID- 21623717 TI - [Functional screening test for latent arterial hypertension detection in young patients]. AB - We assessed diagnostic value of acute-hypoxia test (AHT) for detection of latent arterial hypertension in 317 young persons from different social groups (mean age 25.6+/-5.1 yrs) without clinical signs of cardiovascular diseases. We performed AHT and ambulatory blood pressure monitoring (ABPM). Sensitivity and specificity of AHT for detection 'of latent arterial hypertension were 97.9 and 94.7%, respectively. Agreement of results of AHT and ABPM was 79.6% in patients with high normal blood pressure (BP) and 100% in patients with BP >140/90mm Hg. The test was safe and its results were highly reproducible. PMID- 21623718 TI - [Indices of static and dynamic components of pressure load (assessed by 24-hour blood pressure monitoring) and the state of renal function in patients with essential hypertension]. AB - Aim of this study was to evaluate possible relationship between parameters of blood pressure (BP) profile and glomerular filtration rate in patients (pts) with I-II stage essential hypertension (EH). Material and methods. We studied 120 pts (97 men), aged 23-65 (50,2+/-0,6) years with I (n=98) and II (n=22) stage EH. In BP profile (SL-90207) we calculated 24-hour, daytime, nighttime values of systolic, diastolic, pulse pressures (SBP, DBP, PP), time load (TL), variability and nocturnal fall (NF) of BP. The state of renal function was assessed by measurement of glomerular filtration rate (GFR) calculated by the Cockcroft formula. Results. After nonlinear statistical analysis by Gauss-Newton all patients were divided into three groups according to GFR tertiles. Significant differences were found between these groups by 24-hour, nighttime and daytime values of SBP and DBP. Values of SBP were the lowest in group II. In group II lowest values of PP were also observed, but statistically significant differences were found only in nocturnal PP values between groups II and III. There were no significant differences between groups by TL and NF of BP. In group Ill (high GFR) variability of daytime values of SBP and DBF were significantly higher. Univariate correlation analysis showed statistically significant negative relationship between GFR and nocturnal PP in patients with lowest level of GFR. Positive correlations between nocturnal values of PP and GFR in groups II and III were also observed. Conclusion. These results indicated the presence of strong relationship between high values of nocturnal PP and decreasing of glomerular filtration rate in patients with EH and thus confirmed significance of "constant" and "dynamic" components of pressure load as a marker of impairment of renal function. PMID- 21623719 TI - [Effect of carvedilol and metoprolol R administered with or without atorvastatin on elastic properties of vascular wall and parameters of inflammation in patients with chronic heart failure of ischemic origin]. PMID- 21623720 TI - [Wall motion abnormalities and hemodynamic parameters in patients with left bundle branch block during exercise echocardiography]. AB - Study aim was to investigate dynamics of local contractility and hemodynamic parameters during exercise stress echocardiography (EEcho) in patients with left bundle branch block (LBBB). We examined 23 patients (15 men, 8 women) aged 48-65 years (mean age 53.9+/-8.1 years). Bicycle EEcho was accomplished according to standard protocol. Patients without clinical signs of ischemic heart disease (n=11) comprised group 1, patients after myocardial infarction (n=12) - group 2 (subgroup 2A - with negative test result, subgroup 2B - with worsening of local left ventricular contractility during EEcho). At baseline group 1 patients had significantly better hemodynamic parameters (ejection fraction - EF, left ventricular end diastolic volume - LVEDV) and no abnormalities of local contractility. Exercise tolerance was also the highest in this group. Insignificant worsening of postexercise EF occurred in patients of subgroup 2B (from 46+/-10.5 to 44,2+/-9.4%). In group 1 EF significantly increased (from 56.8+/-10.5% to 64.7+/-15.4%, <0.05), in subgroup 2A tendency to EF increase up to 48.7+/-9.9% was registered. Lowering of local contractility abnormalities index was noted also only in patients of subgroup 2B (from 1.54+/-0.4 to 2.17+/ 0.37 (p<0.01). LVEDV compared with initial values tended to decrease in both groups (however differences between groups were not significant). Positive echocardiographic response was associated with significant changes of transmitral blood flow. Angiographically clean coronary arteries were found in 8 of 10 patients in group 1. Six group 2 patients with history of typical clinical picture of angina and myocardial infarction) had multivessel lesions in coronary vascular bed. EEcho result was positive in 5 of 6 group 2 patients. Thus EEcho possesses high potential for diagnosis of coronary atherosclerosis in patients with LBBB. This allows recommending it as a first line method in patients with this pathology. PMID- 21623721 TI - [Stabilization of cellular membranes as a target of vascular therapy]. AB - We present data of the study of effect of trimetazidine MB on the state of cellular membranes assessed on lymphocytes as highly sensitive model of hypoxia and endothelial dysfunction. Detection of external cytoplasmatic membrane blebbing and registration of quantity of membrane released microparticles by the method of phase-contrast microscopy were carried out in groups of patients with hypertensive disease without and with ischemic heart disease (IHD). It was found that number of cells in the state of terminal blebbing statistically significantly decreased and number of cells with intact cytoplasmic membrane increased at the background of intake of trimetazidine in all groups of patients. With correlation coefficient 0.76 (p<0.05) between content of membrane microparticles and terminal blebbing lowering of content of membrane released microparticles was noted in peripheral blood of patients of all studied groups. The investigation conducted allows to make conclusion that stabilization of cellular membranes is one of spectra of trimetazidine actions underlying improvement of self feeling of patients with vascular pathology. PMID- 21623722 TI - [XVII Congress of the European Society of Cardiology: short report and review of large clinical studies]. PMID- 21623723 TI - [Torasemide is the effective loop diuretic for long-term therapy of arterial hypertension]. AB - Torasemide is a loop diuretic and has been used for the treatment of both acute and chronic congestive heart failure (CHF) and arterial hypertension (AH). Torasemide is similar to other loop diuretics in terms of its mechanism of diuretic action. It has higher bioavailability (>80%) and a longer elimination half-life (3 to 4 hours) than furosemide. In the treatment of CHF torasemide (5 to 20 mg/day) has been shown to be an effective diuretic. Non-diuretic dosages (2.5 to 5 mg/day) of torasemide have been used to treat essential AH, both as monotherapy and in combination with other antihypertensive agents. When used in these dosages, torasemide lowers diastolic blood pressure to below 90mm Hg in 70 to 80% of patients. Antihypertensive efficacy of torasemide is similar to that of thiazides and related compounds. Thus low-dose torasemide constitutes an alternative to thiazides in the treatment of essential AH. PMID- 21623724 TI - [Possibilities of the use of moxonidine in the treatment of arterial hypertension in patients with metabolic syndrome and diabetes]. AB - In pathogenesis of arterial hypertension (AH) and metabolic syndrome (MS) important role plays activation of sympathetic part of vegetative nervous system (VNS). Centrally acting antihypertensive drugs promote lowering of tone of its nuclei. Moxonidine belongs to this group of antihypertensive preparations. Mechanism of action of this drug, peculiarities of its pharmacokinetics are considered in this review. Data on antihypertensive efficacy of moxonidine, possibilities of combination therapy in AH are presented. Literature data on effect of therapy with moxonidine on carbohydrate metabolism, sensitivity of tissues to insulin are presented as well. Possible benefit from administration of this drug to patients with excessive body mass, MS, diabetes mellitus are shown. Data on organoprotective properties of moxonidine (effect on endothelial function, microalbuminuria) are analyzed. PMID- 21623725 TI - [Models of organization of work with patients receiving warfarin for prevention of thromboembolic complications]. PMID- 21623726 TI - [Resident cardiac stem cells]. AB - The search for sources of stem/progenitor cells the use of which has a potential to affect course of ischemic heart disease and chronic heart failure is conducted nowadays in many countries. Resident cardiac stem cells (CSC) were revealed during recent years on the basis of expression of c-kit, sca-1, MDR1, and islet-1 markers. In vitro experiments demonstrated possibility of their differentiation into cardiomyocytes, smooth muscle cell and endothelial cells. Introduction of CSC in injured myocardium in animals facilitated its partial repair and short term improvement of cardiac function. This holds promise for the use of these cells in the future. In the review we have attempted to summarize literature data on resident CSC and their application for the treatment of heart diseases. PMID- 21623727 TI - [Appearance of ECG signs of the Brugada syndrome during therapy with class 1C antiarrhythmic drug ethacizine]. PMID- 21623728 TI - Identification of the novel HLA-B allele B*57:29 by cloning and sequencing in a Chinese individual. AB - HLA-B plays an important role in unrelated donor hematopoietic stem cell transplantation. PMID- 21623729 TI - New single nucleotide polymorphisms were discovered during the psoriasis vulgaris case-control study of gene HLA-C in Shaanxi Chinese Han. PMID- 21623730 TI - Two novel HLA-A alleles: HLA-A*31:01:09 and HLA-A*33:30. AB - The HLA-A*31:01:09 differs from the closest allele HLAA*31:01:02 by a C to T silent substitution at position 447 in exon 3. The HLA-A*33:30 differs from the closest allele HLA-A*33:03:01 by a single nucleotide substitution at position 453 in exon 3, leading to a change of Arg 131 Cys. PMID- 21623731 TI - Identification of a novel allele HLA-DRB1*08:41 in a Chinese donor. AB - HLA-DRB1*08:41 shows seven nucleotide differences from DRB1*08:18 and result in three amino acid changes. PMID- 21623732 TI - Sequencing-based typing identifies three new HLA alleles: C*02:29, C*06:29 and DQB1*03:24. AB - Two new HLA-C alleles, C*02:29 and C*06:29, and one new HLA-DQB1 allele, DQB1*03:24, are described. PMID- 21623733 TI - Association of IL-12p40 +1188 A/C polymorphism with nasopharyngeal cancer risk and tumor extension. AB - The interleukin 12 (IL-12) cytokine, encoded by polymorphic genes, plays a central role in the T helper 1 cell-mediated immunity against tumors. We investigated whether the 3' untranslated region +1188 A/C polymorphism (rs 3212227) influences the nasopharyngeal carcinoma (NPC) risk in Tunisian patients. DNA analysis of 247 patients and 284 healthy individuals showed a higher frequency of the 1188 C allele and the CC genotype in patients than in controls (P = 0.00001 and P = 0.00005) suggesting that the C variant allele is associated with the susceptibility to NPC. Additional testing showed that the homozygous CC genotype is also associated with advanced stage of the tumor extension at presentation (P = 0.022). Our data suggest that the impaired production of IL-12 behaves as a risk factor for NPC occurrence and progression. PMID- 21623734 TI - Breed-independent toll-like receptor 5 polymorphisms show association with canine inflammatory bowel disease. AB - Inflammatory bowel disease (IBD) is thought to be the most common cause of vomiting and diarrhoea in dogs. Although IBD can occur in any canine breed, certain breeds are more susceptible. We have previously shown that polymorphisms in the TLR4 and TLR5 (toll-like receptor) genes are significantly associated with IBD in German Shepherd dogs (GSDs). In order to allow for the development of novel diagnostics and therapeutics suitable for all dogs suffering from IBD, it would be useful to determine if the described polymorphisms are also significantly associated with IBD in other breeds. Therefore, the aim of this study was to investigate whether polymorphisms in the canine TLR4 and TLR5 genes are associated with IBD in other non-GSD canine breeds. The significance of the previously identified non-synonymous single nucleotide polymorphisms (SNPs) in the TLR4 (T23C, G1039A, A1571T and G1807A) and TLR5 genes (G22A, C100T and T1844C) were evaluated in a case-control study using a SNaPSHOT multiplex reaction. Sequencing information from 85 unrelated dogs with IBD consisting of 38 different breeds was compared with a breed-matched control group consisting of 162 unrelated dogs. Indeed, as in the GSD IBD population, the two TLR5 SNPs (C100T and T1844C) were found to be significantly protective for IBD in other breeds (P = 0.023 and P = 0.0195 respectively). Our study suggests that the two TLR5 SNPs, C100T and T1844C could play a role in canine IBD as these were found to be protective factors for this disease in 38 different canine breeds. Thus, targeting TLR5 in the canine system may represent a suitable way to develop new treatment for IBD in dogs. PMID- 21623735 TI - A new method for typing bovine major histocompatibility complex class II DRB3 alleles by combining two established PCR sequence-based techniques. AB - Recently, two polymerase chain reaction sequence-based typing (PCR-SBT) methods were reported for the genotyping of the bovine leukocyte antigen (BoLA)-DRB3. One technique is a single PCR-SBT (sPCR-SBT) method that generates heterozygous sequences that are subsequently analyzed by the haplofinder program, while the other technique is a nested PCR-SBT (nPCR-SBT) method that allows the analysis of heterozygous sequences using the assign 400ATF software. In this study, these techniques were compared and then integrated to produce an improved genotyping method. The primer set used for sPCR-SBT was more accurate than those used for nPCR-SBT. Combining sPCR-SBT with the assign 400ATF software previously reported for nPCR-SBT enables rapid and accurate genotyping of a large number of DNA samples. PMID- 21623736 TI - Lack of KIR2DL4 gene in a fertile Caucasian woman. AB - Killer cell immunoglobulin-like receptor (KIR2DL4) gene is present in virtually all humans. It encodes a receptor present on uterine and decidual natural killer (NK) cells and some peripheral blood NK cells. Its only known ligand is human leukocyte antigen-G molecule expressed on extravillous trophoblasts invading the decidua. Therefore, KIR2DL4 has been regarded as a molecule important for successful pregnancy. However, a multiparous woman from Africa, lacking KIR2DL4 gene, was described suggesting that this gene is not absolutely required for successful human reproduction. Here, we describe a Polish woman who delivered a child and who is not only lacking KIR2DL4 gene, but also possessing a KIR genotype virtually identical to that of the African woman mentioned above. Their genotypes are compared with few other KIR2DL4-negative genotypes and haplotypes described so far. PMID- 21623737 TI - Identification of the new HLA-A*31:48 allele in an Italian patient. AB - Human leukocyte antigen (HLA) class I sequence-based typing (SBT) for hematopoietic unrelated donor searching in an Italian Caucasian patient showed the presence of a novel HLA-A allele defined as A*31:48. HLA-A*31:48 has one nucleotide change from A*31:01:02 at nt 727 from C to T, resulting in an amino acid change at codon 219 of exon 4 from Arg to Trp. PMID- 21623738 TI - Low gestational weight gain and the risk of preterm birth and low birthweight: a systematic review and meta-analyses. AB - BACKGROUND: Low gestational weight gain is common, with potential adverse perinatal outcomes. OBJECTIVE: To determine the relation between low gestational weight gain and preterm birth and low birthweight in singletons in developing and developed countries. DATA SOURCES: Medline, EMBASE and reference lists were searched, identifying 6,283 titles and abstracts. METHODS OF STUDY SELECTION: Following the MOOSE consensus statement, two assessors independently reviewed titles, abstracts, full articles, extracted data and assessed quality. RESULTS: Fifty-five studies, 37 cohort and 18 case-control, were included, involving 3,467,638 women. In the cohort studies (crude data, generally supported where available by adjusted data and case-control studies), women with low total gestational weight gain had increases in preterm birth <37 weeks [RR 1.64 (95%CI 1.62-1.65)], 32-36 weeks [RR 1.39 (95%CI 1.38-1.40)] and <= 32 weeks [RR 3.80 (95%CI 3.72-3.88)]. Low total gestational weight gain was associated with increased risks of low birthweight <2,500 g [RR 1.85 (95%CI 1.72-2.00)], in developing and developed countries [RR 1.84 (95%CI 1.71-1.99) and RR 3.02 (95%CI 1.37-6.63), respectively], 1,500-2,500 g [RR 2.02 (95%CI 1.88-2.17)] and <1,500 g (RR 2.00 (95%CI 1.67-2.40)]. Women with low weekly gestational weight gain were at increased risk of preterm birth [RR 1.56 (95%CI 1.26-1.94)], 32-36 weeks [RR 2.43 (95%CI 2.37-2.50)] and <= 32 weeks [RR 2.31 (95%CI 2.20-2.42)] but not low birthweight [RR 1.64 (95%CI 0.89-3.02)]. CONCLUSIONS: In this systematic review, we determined that singletons born to women with low total gestational weight gain have higher risks of preterm birth and low birthweight, with the lower the gain, the higher the risks. PMID- 21623739 TI - Use of vaginal hysterectomy in Denmark: rates, indications and patient characteristics. AB - OBJECTIVE: To describe the use of vaginal, abdominal and laparoscopic hysterectomy in Denmark from 1999 to 2008, the influence of national guidelines and the patient and procedure-related characteristics associated with the choice of vaginal hysterectomy. Design. Nationwide register-based cohort study. SETTING: Danish Hysterectomy Database and Danish National Patient Registry. POPULATION: All women with a hysterectomy for benign indications in Denmark from 1999 to 2008. METHODS: The incidence rate/100,000 women was used to describe the route of surgery in hysterectomies of 50,755 women. A multiple logistic regression analysis was done to examine the association between patient- and procedure related characteristics and choice of surgical procedure including 20,486 women. MAIN OUTCOME MEASURES: Trends in surgical approach from 1999 to 2008. Patient- and surgery-related characteristics associated with vaginal hysterectomy from 2004 to 2008. RESULTS: There was an overall increase in the use of vaginal hysterectomies from 12 to 34%, a decrease in the use of abdominal hysterectomies and a consistent number of laparoscopic hysterectomies. The number of vaginal hysterectomies varied between regions, ranging from 2 to 86%. The use of vaginal hysterectomy was not dependent on the total number of hysterectomies performed at the hospital. The characteristics associated with vaginal hysterectomy were higher age, smaller uterus size, indications for surgery (genital prolapse and severe uterine bleeding), less smoking and moderate alcohol intake. CONCLUSIONS: Vaginal hysterectomy has replaced abdominal hysterectomy increasingly but cannot be directly correlated to the implementation of national guidelines as there was large national variation. Several characteristics are significantly associated with vaginal hysterectomy. PMID- 21623740 TI - Recurrent early pregnancy loss and consanguinity in Omani couples. AB - Consanguinity is common in Arab countries. The Sultanate of Oman has a relatively small population with a high prevalence of consanguineous marriages. This is a retrospective study of women who had three or more consecutive miscarriages between January 2002 and December 2008, investigated in the non-pregnant state. Age, parity, menstrual history, number of miscarriages, personal and family history, history of consanguinity and investigations were collected for 141 patients. The mean number of miscarriages was 3.4; 53% of the women had a consanguineous marriage, 42% were non-consanguineous and in 5% the marital interrelationship was unknown. There was no significant difference in the prevalence of parental karyotype abnormalities, anticardiolipin antibodies, antinuclear antibodies, thyroid disorders, or lupus between the consanguineous and non-consanguineous couples. Consanguinity appeared not to play a significant role in the etiology of recurrent spontaneous miscarriage. PMID- 21623741 TI - Maternal understanding of commercial cord blood storage for their offspring - a survey among pregnant women in Hong Kong. AB - OBJECTIVE: To assess the knowledge on commercial cord blood banking (CCBB) among pregnant women. DESIGN: Cross-sectional survey. Setting. Antenatal clinics of two major public maternity units in Hong Kong. POPULATION: Pregnant women. METHODS: Self-administered questionnaire. MAIN OUTCOME MEASURES: The survey explored knowledge about the use of self-stored umbilical cord blood (UCB) stem cells and attitude towards CCBB. RESULTS: Of the 2,000 women recruited, 1 866 (93.3%) completed the questionnaire. The majority (78.2%) had no idea that there was the chance of using self-stored stem cells. Moreover, most of the respondents were unclear about which diseases other than leukemia are amenable to treatment with UCB stem cells in general. Only 20.3% of women knew that stem cells are available from the Red Cross in case their children need hematopoietic cell transplantation. CONCLUSIONS: The results of this study revealed inadequate knowledge on UCB stem cell banking and its applications among most of our pregnant women. The government and clinicians should combine efforts to provide accurate information on utilization of UCB stem cells during antenatal care. PMID- 21623742 TI - Mothers' attitudes towards perinatal autopsy after stillbirth. AB - We investigated mothers' attitudes to autopsy of their stillborn baby and their experiences concerning information and treatment in relation to their loss in an observational study. Data were collected by postal questionnaires and telephone calls. Fifty-four of 72 mothers (76%) replied. Fifty-one (94%) received information from a physician about the possibility of having an autopsy; three (6%) did not get any information. The autopsy rate was 83% (n= 45). Thirty-six of 45 (80%) received adequate information about results. Twenty-five (56%) were pleased with how results were presented. Eleven (24%) were positive about individual contact with the pathologist who performed the autopsy. Fifty-one (94%) stated that their decision concerning autopsy was right. Mothers do not regret their decision concerning perinatal autopsy but they do not always receive thorough and timely information concerning autopsy and its results. Personal contact with the perinatal pathologist might help with specific questions both before and after autopsy. PMID- 21623743 TI - Swedish randomized controlled trial of cardiotocography only versus cardiotocography plus ST analysis of fetal electrocardiogram revisited: analysis of data according to standard versus modified intention-to-treat principle. AB - OBJECTIVE: To undertake a renewed analysis of data from the previously published Swedish randomized controlled trial on intrapartum fetal monitoring with cardiotocography (CTG-only) vs. CTG plus ST analysis of fetal electrocardiogram (CTG+ST), using current standards of intention-to-treat (ITT) analysis and to compare the results with those of the modified ITT (mITT) and per protocol analyses. METHODS: Renewed extraction of data from the original database including all cases randomized according to primary case allocation (n=5 049). MAIN OUTCOME MEASURE: Metabolic acidosis in umbilical artery at birth (pH <7.05, base deficit in extracellular fluid >12.0 mmol/l) including samples of umbilical vein blood or neonatal blood if umbilical artery blood was missing. RESULTS: The metabolic acidosis rates were 0.66% (17 of 2 565) and 1.33% (33 of 2 484) in the CTG+ST and CTG-only groups, respectively [relative risk (RR) 0.50; 95% confidence interval (CI) 0.28-0.88; p=0.019]. The original mITT gave RR 0.47, 95%CI 0.25 0.86 (p=0.015), mITT with correction for 10 previously misclassified cases RR 0.48, 95%CI 0.24-0.96 (p=0.038) and per protocol analysis RR 0.40, 95%CI 0.20 0.80 (p=0.009). The level of significance of the difference in metabolic acidosis rates between the two groups remained unchanged in all analyses. CONCLUSION: Re analysis of data according to the ITT principle showed that regardless of the method of analysis, the Swedish randomized controlled trial maintained its ability to demonstrate a significant reduction in metabolic acidosis rate when using CTG+ST analysis for fetal surveillance in labor. PMID- 21623744 TI - Cost-of-illness of patients with chronic hand eczema in routine care: results from a multicentre study in Germany. AB - BACKGROUND: It is broadly assumed that costs caused by chronic hand eczema (CHE) are significant. However, there is a lack of cost-of-illness studies on CHE. OBJECTIVES: To determine the direct and indirect costs of CHE under routine conditions in Germany from the societal perspective. METHODS: A cross-sectional survey was conducted in 24 outpatient practices and clinics across Germany. Patients with CHE refractory to potent topical steroids and insured by statutory health insurance were eligible. Clinical and cost data were collected using standardized questionnaires. Severity classes were defined according to a photographic guide and physician global assessment (PGA). Four treatment stages were defined based on the German CHE guidelines: topical treatments only (stage I), additionally ultraviolet (UV) radiation therapy (II), systemic therapy (III) and inpatient treatment (IV). Bivariate associations between costs and severity as well as treatment stage were assessed. RESULTS: Two hundred and twenty-three patients with CHE (mean age 45.7years, 56% women) enrolled in the study, of whom 63.2% were treated only with topical treatments, 15.7% additionally with UV radiation and 11.7% with systemic treatments. Of all patients, 9.4% had been admitted to hospital. Total costs per year and patient were ?2128, including ?1742 direct costs and ?386 indirect costs. The total costs increased with treatment stages I-IV (P<0.001): ?1044, ?2307, ?2697 and ?8407, respectively. Accordingly, costs also correlated with clinical severity. CONCLUSIONS: Patients with CHE refractory to topical steroids incur marked costs to society. The costs increase disproportionately with escalating treatment stages, especially in patients admitted to hospital. Hence, new treatments may help to reduce the societal costs of CHE. PMID- 21623745 TI - Two novel recessive mutations in KRT14 identified in a cohort of 21 Spanish families with epidermolysis bullosa simplex. AB - BACKGROUND: Basal epidermolysis bullosa simplex (EBS) is a group of blistering genodermatoses mostly caused by mutations in the keratin genes, KRT5 and KRT14. Recessive mutations represent about 5% of all EBS mutations, being common and specific in populations with high consanguinity, where affected patients show severe phenotypes. OBJECTIVES: To accomplish the first mutational analysis in patients of Spanish origin with EBS and to delineate a comprehensive genotype phenotype correlation. METHODS: Twenty-one EBS families were analysed. Immunofluorescence mapping at the dermoepidermal junction level was performed on skin biopsies from patients. Mutation screening of the entire coding sequences of KRT5 and KRT14 in genomic DNA was assessed by polymerase chain reaction and direct sequencing. RESULTS: KRT5 or KRT14 causative mutations were identified in 18 of the 21 EBS families. A total of 14 different mutations were disclosed, of which 12 were dominant missense mutations and two truncating recessive mutations. Five of the 14 mutations were novel including three dominant in KRT5 (p.V186E, p.T321P and p.A428T) and two recessive in KRT14 (p.K116X and p.K250RfsX8). The two patients with EBS carrying homozygous recessive mutations were affected by severe phenotypes and belonged to consanguineous families. All five families with the EBS Dowling-Meara subtype carried recurrent mutations affecting the highly conserved ends of the alpha-helical rod domain of K5 and K14. The seven mutations associated with the localized EBS subtype were widely distributed along the KRT5 and KRT14 genes. Two families with mottled pigmentation carried the P25L mutation in KRT5, commonly associated with this subtype. CONCLUSIONS: This study further confirms the genotype-phenotype correlation established for EBS in other ethnic groups, and is the first in a Mediterranean country (excluding Israel). This study adds two novel recessive mutations to the worldwide record to date, which includes a total of 14 mutations. As in previous reports, the recessive mutations resulted in a lack of keratin K14, giving rise to a generalized and severe presentation. PMID- 21623746 TI - Parameters associated with severe pain during photodynamic therapy: results of a large Scottish series. PMID- 21623747 TI - Sun exposure rapidly reduces plasmacytoid dendritic cells and inflammatory dermal dendritic cells in psoriatic skin. AB - BACKGROUND: Interferon (IFN)-alpha-producing plasmacytoid dendritic cells (pDCs), inflammatory CD11c+CD1c- myeloid dendritic cells (mDCs) and macrophages have been found to contribute to the pathogenesis of psoriasis. Heliotherapy is a well established treatment modality of this disease, although the details of how the effects are mediated are unknown. OBJECTIVES: To test the hypothesis that exposure to natural sun affects pathogenic DC subsets in lesional skin. METHODS: Skin biopsies were obtained from lesional and nonlesional skin in 10 patients with moderate to severe psoriasis subjected to controlled sun exposure on Gran Canaria. Biopsies were obtained at baseline, day 2 and day 16 and examined by immunohistochemistry. RESULTS: Sixteen days of heliotherapy had excellent clinical effect on patients with psoriasis, with significant reductions in Psoriasis Area and Severity Index (PASI) scores. In lesional skin pDC numbers and expression of MxA, a surrogate marker for IFN-alpha, were rapidly reduced. Inflammatory CD11c+CD1c- mDCs were significantly reduced whereas resident dermal CD11c+CD1c+ mDCs were unaffected. Expression levels of the maturation marker DC LAMP (CD208) on mDCs were significantly reduced after sun exposure, as were the numbers of lesional dermal macrophages. A decrease of dermal DC subsets and macrophages was already observed after 1 day of sun exposure. An additional finding was that DC-SIGN (CD209) is primarily expressed on CD163+ macrophages and not DCs. CONCLUSIONS: The clinical improvement in psoriasis following sun exposure is associated with rapid changes in dermal DC populations and macrophages in lesional skin, preceding the clinical effect. These findings support the concept that these DC subsets are involved in the pathogenesis of psoriasis and suggest that sun-induced clinical benefit may partly be explained by its effect on dermal DCs. PMID- 21623748 TI - A modified, improved, easy and fast technique for split-thickness skin grafting. AB - BACKGROUND: Large nonhealing ulcers and wounds frequently pose a great therapeutic challenge to clinicians and often require skin grafting. Various skin grafting methods are available to cover large skin defects that fail to epithelize. These methods include the use of small pinch grafts, full-thickness punch grafts, large-sized full-thickness grafts and split-thickness grafts. Large sized full-thickness and split-thickness skin grafting requires expertise to produce cosmetically acceptable results and prevent cobblestoning, unlike small pinch and full-thickness punch grafts. OBJECTIVES: To describe a modified technique of split-thickness skin grafting that can be considerably faster than alternative methods. METHODS: We describe a method for split-thickness skin grafting using tumescent anaesthesia at the donor site and an electrodermatome and a polyurethane membrane without sutures at the site of the skin defect. RESULTS: Since 1997, we have practised a modified, improved, quick and easy split thickness skin grafting method to cover large skin defects at the extremities. Complete healing is usually achieved 4-6 weeks after the split-thickness skin transplantation, and long-term results are aesthetically successful. CONCLUSIONS: We provide a sophisticated modified split-thickness skin graft procedure that has been practised for many years and provides cosmetically acceptable results while saving time. PMID- 21623749 TI - Blinded vs. unblinded peer review of manuscripts submitted to a dermatology journal: a randomized multi-rater study. AB - BACKGROUND: Submissions to medical and scientific journals are vetted by peer review, but peer review itself has been poorly studied until recently. One concern has been that manuscript reviews in which the reviewer is unblinded (e.g. knows author identity) may be biased, with an increased likelihood that the evaluation will not be strictly on scientific merits. OBJECTIVES: The purpose of this study was to compare the outcomes of blinded and unblinded reviews of manuscripts submitted to a single dermatology journal via a randomized multi rater study. MATERIALS AND METHODS: Forty manuscripts submitted to the journal Dermatologic Surgery were assessed by four reviewers, two of whom were randomly selected to be blinded and two unblinded regarding the identities of the manuscripts' authors. The primary outcome measure was the initial score assigned to each manuscript by each reviewer characterized on an ordinal scale of 1-3, with 1 = accept; 2 = revise (i.e. minor or major revisions) and 3 = reject. Subgroup analysis compared the primary outcome measure across manuscripts from U.S. corresponding authors and foreign corresponding authors. The secondary outcome measure was word count of the narrative portion (i.e. comments to editor and comments to authors) of the reviewer forms. RESULTS: There was no significant difference between the scores given to manuscripts by unblinded reviewers and blinded reviewers, both for manuscripts from the U.S. and for foreign submissions. There was also no difference in word count between unblinded and blinded reviews. CONCLUSIONS: It seems, at least in the case of one dermatology journal, that blinding during peer review does not appear to affect the disposition of the manuscript. To the extent that review word count is a proxy for review quality, there appears to be no quality difference associated with blinding. PMID- 21623750 TI - Topical aminolaevulinic acid-photodynamic therapy produces an inflammatory infiltrate but reduces Langerhans cells in healthy human skin in vivo. AB - BACKGROUND: Topical photodynamic therapy (PDT) elicits a therapeutic response in both skin cancer and immune-mediated skin disorders. While PDT induces direct cell death, host inflammatory and immune responses to PDT may contribute to the therapeutic effects. OBJECTIVES: To examine the impact of topical PDT on leucocyte trafficking and mediators of chemotaxis in healthy human skin. METHODS: Aminolaevulinic acid (ALA)-PDT was performed on the buttock skin of seven healthy volunteers. Biopsies for immunohistochemical assessment were taken 1, 4 and 24 h post-PDT and from untreated contralateral buttock skin (baseline). RESULTS: A significant dermal neutrophilic infiltrate appeared early, peaking at 4 h (P < 0.01) and returning to near baseline by 24 h. Expression of E-selectin was significantly higher at 4 h (P < 0.05) and correlated strongly with neutrophil numbers (r = 0.93). Expression of intercellular adhesion molecule 1 was significantly elevated after 24 h (P < 0.05) with an apparent gradual increase in CD4+ T cells up to this time point. Notably, epidermal Langerhans cells were significantly reduced 24 h post-PDT compared with baseline (P < 0.01) and comprised a significantly larger proportion of cells with migratory rather than dendritic morphology (P < 0.05). The number of epidermal cells expressing tumour necrosis factor-alpha significantly increased at 4 h (P < 0.05) and remained elevated 24 h post-PDT, whereas no significant change in expression of interleukin (IL)-1beta or IL-8 was seen. CONCLUSIONS: Reduction of Langerhans cells by topical PDT of human skin may play a significant role in PDT-induced local immunosuppression, potentially benefiting the treatment of immune-mediated skin disorders but negatively impacting on antitumour responses. Further exploration according to disease indication/treatment protocol is warranted. PMID- 21623752 TI - High-concentration all-trans retinoic acid induces dermal inflammation and reduces the accumulation of type I procollagen in human skin in vivo. AB - BACKGROUND: Because inflammation is a factor promoting ageing, all-trans retinoic acid (RA)-induced irritation may have a negative influence on collagen accumulation in human skin despite its stimulation of collagen production. OBJECTIVES: To determine whether RA-induced irritation detrimentally affects RA efficacy as represented by new collagen synthesis. METHODS: Retinoic acid (0.01%, 0.025% or 0.05%) or vehicle was applied to the buttock skin of elderly male volunteers three times a week for 8 weeks under continuous occlusion. Every 2 weeks, biopsy specimens were obtained and immunohistochemical analysis was performed to determine levels of type I procollagen expression and inflammatory cell infiltration. RESULTS: Topical RA regardless of concentration increased type I procollagen expression in human skin in vivo after 2 weeks. However, only 0.01% RA continuously increased type I procollagen expression up to 8 weeks. After 4 weeks, significant infiltrations of macrophages and neutrophils were observed in 0.025% and 0.05% RA-treated skin, and procollagen expression had returned to baseline. CONCLUSIONS: Excessive RA-induced inflammation might prevent collagen accumulation in aged skin despite the positive effect of RA on collagen production. PMID- 21623751 TI - Increases in invasive melanoma in England, 1979-2006, by anatomical site. AB - BACKGROUND: National melanoma incidence trends with details of anatomical site have not been previously described for England. OBJECTIVES: To describe site specific trends in cutaneous melanoma for England as a whole during the last three decades. METHODS: Anonymized data, 1979-2006, were obtained from national cancer registrations of all patients in England up to age 89years with incident primary invasive cutaneous melanomas (n=124055). Sex-specific age-standardized incidence rates and average annual percentage change in rates were calculated for each broad anatomical site. RESULTS: Overall incidence rates of cutaneous melanoma in England, 1979-2006, were 81 and 100 per million, in males and females, respectively. Site-specific rates were consistently highest on the lower limbs in females followed by the trunk in males. Greatest annual increases occurred on the trunk in both sexes over 45years (males 9.9%, females 6.8%), then upper limbs (males 8.7%, females 6.8%). Incidence trends in males relative to females varied little across sites apart from a more rapid rise in head/neck melanomas in males than in females after the 1980s. CONCLUSIONS: Invasive melanoma rates continue to rise in England, particularly on the trunk and arms, and in males on the head/neck. The steeper increases in melanoma rates among males are consistent with their greater sun exposure and poorer compliance with sun protection measures than females. PMID- 21623753 TI - Prevalence of skin lesions and need for treatment in a cohort of 90 880 workers. AB - BACKGROUND: Healthcare planning requires robust data on the prevalence and need for care of dermatological diseases. To date, no systematic data in population based samples are available for Germany. OBJECTIVES: Determination of the prevalence of skin lesions and of the need for care based on dermatological examinations in working adults in Germany. METHODS: From 2004 to 2009, workers aged between 16 and 70years from different branches of industry throughout Germany underwent a single dermatological whole-body examination on the occasion of company screening for skin cancer. The data were recorded electronically and evaluated descriptively. In addition to the clinical findings, case history data on previous skin diseases were documented and the need for further clarification or treatment was determined on the basis of the dermatologist's assessment. RESULTS: Data from n=90 880 persons from a total of 312 companies were evaluated. Of the pigmented skin lesions, dermal naevi were found in 25.1% of the cohort, and 16.7% displayed more than 40 each. The most frequent inflammatory skin diseases were acne vulgaris (3.9%), psoriasis (2.0%), rosacea (2.3%) and atopic eczema (1.3%). Examination of the case histories showed that the most frequent condition was allergic sensitizations (41.1%); of these, pollen accounted for the biggest group (21.4%), followed by contact allergens (8.0%). In total, 26.8% of the cohort exhibited a dermatological finding in need of treatment or further clarification. CONCLUSIONS: Dermatological lesions and diseases requiring clarification are frequent and indicate a high demand for treatment in the adult population. PMID- 21623754 TI - Paratumoral gene expression profiles: promising markers of malignancy in melanocytic lesions. PMID- 21623755 TI - The mesenchymal stem cell profile in psoriasis. AB - BACKGROUND: The expression of inducible nitric oxide synthase (iNOS) and vascular endothelial growth factor (VEGF) and the level of total oxyradical scavenging capacity have been evaluated extensively in the cutaneous cells of patients with psoriasis. As yet, no indications are available about the undifferentiated cells, the mesenchymal stem cells (MSCs), isolated from skin. OBJECTIVES: To isolate MSCs in patients with psoriasis and to compare them with those obtained from atopic and healthy subjects, in order to analyse whether MSCs show some typical psoriatic profiles and to understand whether pathophysiological events leading to psoriasis start early at the stem cell level. METHODS: MSCs isolated from seven patients with psoriasis, seven patients with acute atopic dermatitis and seven healthy subjects were characterized by fluorescence-activated cell sorting analysis. VEGF and nitric oxide (NO) content was measured in conditioned medium, the expression of VEGF and iNOS was analysed by immunohistochemistry, and the total oxyradical scavenging capacity towards peroxynitrite was tested. RESULTS: VEGF content was highest in the medium conditioned by psoriatic perilesional MSCs, whereas NO concentration was maximally increased in medium conditioned by MSCs isolated from lesional psoriatic skin. The ability to neutralize the oxidizing effects of peroxynitrite was lower for MSCs isolated from lesional psoriatic skin compared with other MSCs, except for MSCs of lesional atopic skin. CONCLUSIONS: The microenvironment in psoriasis differs from those of atopic dermatitis and healthy skin; it could induce resident MSCs to produce angiogenic and proinflammatory mediators which lead to a reduction in the antioxidant capacity of these cells, contributing to the development of skin lesions in psoriasis. PMID- 21623756 TI - Adenovirus-relaxin gene therapy for keloids: implication for reversing pathological fibrosis. AB - BACKGROUND: Keloids or hypertrophic scars are pathological proliferations of the dermal skin layer resulting from excessive collagen deposition. Because the hormone relaxin (RLX) inhibits collagen synthesis and expression in stimulated fibroblasts, an adenovirus expressing RLX (dE1-RGD/lacZ/RLX) was generated. OBJECTIVES: To investigate the effect of RLX-expressing adenovirus on expression of various extracellular matrix (ECM) components in primary keloid spheroids. METHODS: The expression levels of type I and III collagen, fibronectin and elastin were investigated by immunohistochemistry in primary keloid spheroids transduced with the RLX-expressing adenovirus. RESULTS: Immunohistochemical analysis showed that expression of major ECM components (e.g. type I and III collagen, elastin and fibronectin) was markedly reduced in primary keloid spheroids transduced with dE1-RGD/lacZ/RLX. CONCLUSIONS: These results suggest that the antifibrotic effect of RLX-expressing adenovirus may have therapeutic effects on keloids by reversing pathological fibrosis and preventing keloid recurrence after surgical excision. PMID- 21623757 TI - Low-dose isotretinoin in acne vulgaris: a critical review. PMID- 21623758 TI - Outcomes of 11 pregnancies in three patients with recessive forms of epidermolysis bullosa. PMID- 21623759 TI - Pancreatic pseudocyst in an adult patient after treatment with pegylated asparaginase. PMID- 21623760 TI - Recent advances in the pathogenesis and treatment of juvenile myelomonocytic leukaemia. AB - Myeloid neoplasms derive from the pathological clonal expansion of an abnormal stem cell and span a diverse spectrum of phenotypes including acute myeloid leukaemia (AML), myeloproliferative neoplasms (MPN) and myelodysplastic syndromes (MDS). Expansion of myeloid blasts with suppression of normal haematopoiesis is the hallmark of AML, whereas MPN is associated with over-proliferation of one or more lineages that retain the capacity to differentiate, and MDS is characterized by cytopenias and aberrant differentiation. MPD and MDS can progress to AML, which is likely due to the acquisition of cooperative mutations. Juvenile myelomonocytic leukaemia (JMML) is an aggressive myeloid neoplasm of childhood that is clinically characterized by overproduction of monocytic cells that can infiltrate organs, including the spleen, liver, gastrointestinal tract, and lung. JMML is categorized as an overlap MPN/MDS by the World Health Organization and also shares some clinical and molecular features with chronic myelomonocytic leukaemia, a similar disease in adults. While the current standard of care for patients with JMML relies on allogeneic haematopoietic stem cell transplant (HSCT), relapse is the most frequent cause of treatment failure. This review outlines our understanding of the genetic underpinnings of JMML with a recent update on the discovery of novel CBL mutations, as well as a brief review on current therapeutic approaches. PMID- 21623761 TI - AICDA expression in BCR/ABL1-positive acute lymphoblastic leukaemia is associated with a peculiar gene expression profile. AB - Activation-induced cytidine deaminase (AICDA) initiates somatic hypermutation and class-switch recombination of immunoglobulin (Ig) genes and induces mutations also in non-Ig genes. AICDA aberrant expression was detected in B-lineage acute lymphoblastic leukaemia (B-ALL), particularly BCR/ABL1+ B-ALL; patients expressing AICDA carried more copy number alterations than 'AICDA-negative' cases. To determine the role of AICDA, AICDA expression and gene expression profiling were studied in adult BCR/ABL1+ B-ALL. Patients displaying the full length isoform AICDA are characterized by up-regulation of DNA repair/replication and cell cycle genes, suggesting their involvement in the genetic instability of BCR/ABL1+ B-ALL. PMID- 21623762 TI - Comparison of intravenous immunoglobulin and high dose anti-D immunoglobulin as initial therapy for childhood immune thrombocytopenic purpura: response Ozsoylu. PMID- 21623763 TI - Retraction: transactivation of CCL20 gene by Epstein-Barr virus latent membrane protein 1. PMID- 21623764 TI - Retraction: a modified version of galectin-9 induces cell cycle arrest and apoptosis of Burkitt and Hodgkin lymphoma cells. PMID- 21623766 TI - Identification of RHCE and KEL alleles in large cohorts of Afro-Caribbean and Comorian donors by multiplex SNaPshot and fragment assays: a transfusion support for sickle cell disease patients. AB - To lower the alloimmunization risk following transfusion in blacks, we developed two genotyping assays for large-scale screening of Comorian and Afro-Caribbean donors. One was a multiplex SNaPshot assay designed to identify ce(s) (340), ceMO/AR/EK/BI/SM, ce(s) , ce(s) (1006) and KEL*6/*7 alleles. The other was a multiplex fragment assay designed to detect RHD, RHDpsi and RHCE*C and 455A>C transversion consistent with (C)ce(s) Type 1 and DIII Type5 ce(s) . Variant RHCE*ce alleles or RH haplotypes were detected in 58.69% of Comorians and 41.23% of Afro-Caribbeans. The ce(s) allele, (C)ce(s) Type 1, and DIII Type 5 ce(s) haplotypes were identified respectively in 39.13%, 14.67% and 4.88% of Comorians and 32.23%, 5.28% and 1.76% of Afro-Caribbeans. Genotypes consistent with partial D, C, c and/or e antigen expression were observed in 26.08% of Comorians and 14.69% of Afro-Caribbeans. No homozygous genotype corresponding to the RH:-18, 34, and -46 phenotypes were found. However, over 50% of genotypes produced low prevalence antigens at risk for negative recipients, i.e., V, VS, JAL, and/or KEL6. One new variant RHCE*ce(s) (712) allele was identified. This is the first determination of variant RHCE and KEL allele frequencies. Results indicate the most suitable targets for molecular assay screening to optimize use of compatible blood units and lower immunization risk. PMID- 21623767 TI - Malaria: modification of the red blood cell and consequences in the human host. AB - Residence in the human erythrocyte is essential for the lifecycle of all Plasmodium that infect man. It is also the phase of the life cycle that causes disease. Although the red blood cell (RBC) is a highly specialized cell for its function of carrying oxygen to and carbon dioxide away from tissues, it is devoid of organelles and lacks any cellular machinery to synthesize new protein. Therefore in order to be able to survive and multiply within the RBC membrane the parasite needs to make many modifications to the infected RBC (iRBC). Plasmodium falciparum (P. falciparum) also expresses parasite-derived proteins on the surface of the iRBC that enable the parasite to cytoadhere to endothelial and other intravascular cells. These RBC modifications are at the root of malaria pathogenesis and, in this ancient disease of man, have formed the epicentre of a genetic 'battle' between parasite and host. This review discusses some of the critical modifications of the RBC by the parasite and some of the consequences of these adaptations on disease in the human host, with an emphasis on advances in understanding of the pathogenesis of severe and cerebral malaria (CM) from recent research. PMID- 21623768 TI - Increased interleukin-4-positive lymphocytes in patients with Hashimoto's thyroiditis and concurrent non-endocrine autoimmune disorders. AB - A prevalent T helper type 1 (Th1) subset of lymphocytes has been described in Hashimoto's thyroiditis (HT), but whether a similar polarization may characterize HT when associated with non-endocrine autoimmune disorders (NEAD) is not known. The aim of the present study was to analyse the intracellular Th1 and Th2 distinctive cytokines in patients with isolated HT or associated with non endocrine autoimmune disorders. Intracellular cytokine expression was assessed in peripheral blood lymphocytes (PBL) of 68 out-patients (females = 55; males = 13; median age = 6 years) with HT : 33 had isolated HT and 35 had a concurrent NEAD. The percentage of interferon (IFN)-gamma and interleukin (IL)-2 Th1- and IL-4 Th2 positive cells was measured by flow cytometric analysis. We found an increased percentage of IL-2-positive cells in all patients, without differences between patients with isolated HT or associated with NEAD. IFN-gamma(+) cells were also increased in both groups, but the median percentage of those with isolated HT was lower than in patients with HT+NEAD (19.0 versus 29.9%; P = 0.0082). An increased number of IL-4-positive cells was observed in three of 33 (9.1%) patients with isolated HT and in 25 of 35 patients with NEAD [71%; P < 0.0001; relative risk (RR) = 3.18]. The median values of IL-4(+) cells (HT = 5.0% versus HT + NEAD = 16.8%) confirmed this large difference (P < 0.0001). A clear-cut increase of IL 4(+) lymphocytes characterizes patients with autoimmune thyroiditis who have associated non-endocrine autoimmune disorders. These findings may represent an initial tool to detect patients with autoimmune thyroiditis in which additional non-endocrine autoimmune disorders may be awaited. PMID- 21623769 TI - Amplicon-based high-throughput pooled sequencing identifies mutations in CYP7B1 and SPG7 in sporadic spastic paraplegia patients. AB - Hereditary spastic paraplegia (HSP) is a neurodegenerative disorder defined clinically by progressive lower limb spasticity and weakness. HSP is a genetically highly heterogeneous condition with at least 46 gene loci identified so far, involving X-linked, autosomal recessive (AR) and autosomal dominant inheritance. For correct diagnosis, molecular testing is essential because clinical parameters by themselves are not reliable to differentiate HSP forms. The purpose of this study was to establish amplicon-based high-throughput genotyping for AR-HSP. A sample of 187 index cases with apparently sporadic or recessive spastic paraplegia were analyzed by applying an array-based amplification strategy. Amplicon libraries of the CYP7B1-(SPG5) and SPG7-gene were generated followed by a pooled next-generation sequencing (NGS) approach. We identified three SPG5 and seven SPG7 patients. All had one homozygous or two heterozygous mutations. In total, 20 distinct mutations (CYP7B1,n = 4and SPG7,n = 16) including two novel CYP7B1 mutations (p.G51R and p.E211KfsX3) and eight novel SPG7 mutations (p.Leu8delinsLeuLeu, p.W29X, p.R139X, p.R247X, p.G344D, p.Leu346_Leu347ins11, p.R398X and p.R398Q) were detected by this comprehensive genetic testing. Our study illustrates how amplicon-based NGS can be used as an efficient tool to study genotypes and mutations in large patient cohorts and complex phenotypes. PMID- 21623770 TI - Evidence for disease penetrance relating to CNV size: Pelizaeus-Merzbacher disease and manifesting carriers with a familial 11 Mb duplication at Xq22. AB - The potential causes for the incomplete penetrance of Pelizaeus-Merzbacher disease (PMD) in female carriers of PLP1 mutations are not well understood. We present a family with a boy having PMD in association with PLP1 duplication and three females who are apparent manifesting carriers. Custom high-resolution oligonucleotide array comparative genomic hybridization (aCGH) and breakpoint junction sequencing were performed and revealed a familial complex duplication consisting of a small duplicated genomic interval (~56 kb) and a large segmental duplication (~11 Mb) that resulted in a PLP1 copy number variation gain. Breakpoint junction analysis implicates a replication-based mechanism underlying the rearrangement formation. X-inactivation studies (XCI) showed a random to moderate advantageous skewing pattern in peripheral blood cells but a moderate to extremely skewed (>=90%) pattern in buccal cells. In conclusion, our data show that complex duplications involving PLP1 are not uncommon, can be detected at the level of genome resolution afforded by clinical aCGH and duplication and inversion can be produced in the same event. Furthermore, the observation of three manifesting carriers with a large genomic rearrangement supports the contention that duplication size along with genomic content can be an important factor for penetrance of the PMD phenotype in females. PMID- 21623771 TI - Mutations in the motor and stalk domains of KIF5A in spastic paraplegia type 10 and in axonal Charcot-Marie-Tooth type 2. AB - Spastic paraplegia type 10 (SPG10) is an autosomal dominant form of hereditary spastic paraplegia (HSP) due to mutations in KIF5A, a gene encoding the neuronal kinesin heavy chain implicated in anterograde axonal transport. KIF5A mutations were found in both pure and complicated forms of the disease; a single KIF5A mutation was also detected in a CMT2 patient belonging to an SPG10 mutant family. To confirm the involvement of the KIF5A gene in both CMT2 and SPG10 phenotypes and to define the frequency of KIF5A mutations in an Italian HSP patient population, we performed a genetic screening of this gene in a series of 139 HSP and 36 CMT2 affected subjects. We identified five missense changes, four in five HSP patients and one in a CMT2 subject. All mutations, including the one segregating in the CMT2 patient, are localized in the kinesin motor domain except for one, falling within the stalk domain and predicted to generate protein structure destabilization. The results obtained indicate a KIF5A mutation frequency of 8.8% in the Italian HSP population and identify a region of the kinesin protein, the stalk domain, as a novel target for mutation. In addition, the mutation found in the CMT2 patient strengthens the hypothesis that CMT2 and SPG10 are the extreme phenotypes resulting from mutations in the same gene. PMID- 21623772 TI - Fabry disease 'The New Great Imposter': results of the French Observatoire in Internal Medicine Departments (FIMeD). AB - Fabry disease (FD) is an X-linked lysosomal storage disorder due to alpha galactosidase A deficiency. It is associated with a broad range of clinical symptoms, resulting in frequent misdiagnosis and diagnostic delay, which may impact on patient outcomes. This retrospective observational study of 58 FD patients referred to 10 internal medicine departments in France aimed to review differential diagnoses received prior to diagnosis and examines diagnostic delay. The average age at the time of diagnosis was 27.6 years (range: 10-60) and 42.2 years (range: 9-77) among the 23 males and 35 females analyzed, respectively. Most common symptoms that led to FD diagnosis were family history of FD (12 males and 27 females), followed by pain in extremities (10 males and 5 females), and angiokeratoma (8 males and 4 females). Eighteen patients had received alternative diagnoses prior to FD diagnosis, including a female patient with four previous diagnoses. Four case reports are presented, which illustrate the diagnostic 'odyssey' and delayed diagnosis often experienced by patients. Clinicians should consider a diagnosis of FD when presented with a wide range of symptoms, thus helping to shorten the diagnostic delay and facilitating early therapy with enzyme replacement therapy to improve patient outcomes. PMID- 21623773 TI - High-sensitivity C-reactive protein in psoriasis. PMID- 21623774 TI - Subcutaneous phaeohyphomycosis caused by Exophiala jeanselmei treated with wide surgical excision and posaconazole: case report. PMID- 21623775 TI - Syringolymphoid hyperplasia with alopecia and anhidrosis in a 12-year-old boy: a case report from rural south India. PMID- 21623776 TI - The effect of oral isotretinoin (13-cis retinoic acid) on hearing systems in patients with acne vulgaris: a prospective study. AB - Isotretinoin is widely used in the treatment of extensive and nodulocystic acne. The objective of this prospective study was to investigate whether oral isotretinoin could affect the hearing system. Thirty-eight patients with acne vulgaris (76 ears) who were diagnosed and treated at the Department of Dermatology were included in the current study. Study evaluation visits were performed at baseline and at Weeks 1, 2 and 3. Pure-tone averages (PTAs) of air conduction thresholds at 250 Hz (PTA1); 500, 1000, and 2000 Hz (PTA2); 4000, 8000, and 10,000 Hz (PTA3); and 12,500, 16,000, 18,000 and 20,000 Hz (PTA4) for each ear were calculated separately. Assessment of the efficacy was based on the audiometric findings. Compared with pre-treatment evaluation, the PTAs of patients were found to be significantly different at the first week for PTA2 (P = 0.033) and PTA3 (P = 0.001), at the second week for PTA1 (P = 0.036), and at the third week for PTA4 (P = 0.002). Our results suggest that the oral isotretinoin (13-cis retinoic acid), which is a derivative of retinol (vitamin A), improved the hearing level of the patients in all audiometric frequencies in a short period follow-up. PMID- 21623777 TI - Oxidative stress in ischaemic stroke. AB - BACKGROUND: Production of reactive oxygen species after ischaemic stroke may enhance tissue damage through multiple molecular pathways. MATERIALS AND METHODS: In this study, we examined the serum levels of lipoperoxide and hydroperoxide, conjugated dienes and total antioxidant capacity levels in 50 patients with acute ischaemic stroke (T0) to evaluate the possibility to use them as specific biochemical markers for cerebral ischaemia. Determinations were repeated after a month (T1) to correlate their relative changes with clinical evolution. RESULTS: Lipoperoxide, hydroperoxide and conjugated diene levels in platelets were significantly higher in the early stages with respect to their late evaluation. On the contrary, total antioxidant capacity showed a significant increase at T1 with respect to T0. A significant negative correlation between total antioxidant capacity and NIHSS score at T0 and T1 was found. There was a significant positive correlation between lipoperoxide, hydroperoxide and conjugated dienes levels and NIHSS score at T0 and at T1. CONCLUSIONS: These findings suggest that changes in free radical generation and consequent oxidative stress may have a role in the pathogenesis of acute ischaemic lesions. The activation of defence mechanisms like total antioxidant capacity could be involved in the limitation of ischaemic damage progression. PMID- 21623778 TI - Adiponectin in chronic heart failure: influence of diabetes and genetic variants. AB - BACKGROUND: We hypothesized that, besides type 2 diabetes (T2D) and body mass index (BMI), circulating adiponectin concentration would be associated with variants of the ADIPOQ gene in patients with chronic heart failure (CHF). We also assessed the influence of these confounders on the prognostic value of adiponectin. METHODS: Plasma adiponectin was measured at entry and after 3 months in approximately 1200 patients with CHF enrolled in the GISSI-HF trial. Four common single-nucleotide polymorphisms (SNPs) spanning the ADIPOQ gene were studied: rs17300539 (-11391G->A), rs266729 (-11377C->G), rs2241766 (+45T->G) and rs1501299 (+276G->T). Associations with clinical characteristics and mortality were evaluated in patients with or without T2D. RESULTS: Adiponectin concentrations were negatively related to BMI, higher in women and older persons, but lower in patients with diabetes. T-allele carriers for rs1501299 and A-allele carriers for rs17300539 had significantly elevated adiponectin concentrations. Irrespective of diabetes, baseline plasma adiponectin was independently associated with mortality (adjusted HR [95%CI] per 1 SD increase in adiponectin concentration = 1.24[1.12-1.37], P < 0.0001) and improved prognostic discrimination beyond clinical risk factors (integrated discrimination improvement, P = 0.005). Patients with increasing adiponectin concentration over 3 months had worse outcome than those with stable levels (unadjusted HR = 1.46[1.09-1.96], P = 0.01); this relation was attenuated by the genetic variants examined and by robust confounders like age, diabetes, BMI or NT-proBNP (adjusted HR = 1.37[0.97-1.94], P = 0.075). CONCLUSIONS: Although diabetes and genetic variants at the ADIPOQ gene influence the circulating levels of adiponectin in CHF, higher plasma adipokine levels, but not genetic variants, are consistently associated with a poor prognosis. PMID- 21623779 TI - Non polycystic ovary syndrome-related endocrine disorders associated with hirsutism. AB - BACKGROUND: Hyperandrogenism refers to classical androgen-dependent signs such as hirsutism, acne and androgenetic alopecia. Hirsutism is the main hyperandrogenic symptom, defined as an excess of body hair in the androgen-sensitive skin regions of the women. In this review, we attempt to focus on the pathogenesis of hirsutism related to disorders other than polycystic ovary syndrome (PCOS). Also, we will discuss their clinical and biochemical features as well as therapeutic options. DESIGN: Several original articles, meta-analysis and reviews have been screened in the field of hirsutism and hyperandrogenic disorders. RESULTS: Current English literature including our studies suggests that PCOS is the most common cause of hirsutism. The most important purpose for investigation is to identify those women with androgen-secreting tumours because of their life threatening potential. In approximately 1-8% of the women with hirsutism, the underlying cause is nonclassical adrenal hyperplasia because of 21-hydroxylase deficiency. Depending on ethnicity and the geographic area, idiopathic hirsutism constitutes 5-17% of the patients with hirsutism. Approximately 3% of hyperandrogenic women were observed to suffer from hyperandrogenic-insulin resistant acanthosis nigricans syndrome. More rare causes are glucocorticoid resistance syndrome, hyperprolactinemia, acromegaly, Cushing's syndrome and some drugs. Specific causes of hirsutism such as Cushing's syndrome and adrenal/ovarian tumours should be treated specifically. In other patients, pharmacological approach is the mainstay of therapy. CONCLUSIONS: A number of patients presenting with hirsutism and exhibiting similar features to PCOS may have other underlying diagnoses. Unlike PCOS, some of these disorders can occasionally be life threatening and require prompt diagnosis and treatment. PMID- 21623780 TI - Evaluation of MODFLOW-LGR in connection with a synthetic regional-scale model. AB - This work studies costs and benefits of utilizing local-grid refinement (LGR) as implemented in MODFLOW-LGR to simulate groundwater flow in a buried tunnel valley interacting with a regional aquifer. Two alternative LGR methods were used: the shared-node (SN) method and the ghost-node (GN) method. To conserve flows the SN method requires correction of sources and sinks in cells at the refined/coarse grid interface. We found that the optimal correction method is case dependent and difficult to identify in practice. However, the results showed little difference and suggest that identifying the optimal method was of minor importance in our case. The GN method does not require corrections at the models' interface, and it uses a simpler head interpolation scheme than the SN method. The simpler scheme is faster but less accurate so that more iterations may be necessary. However, the GN method solved our flow problem more efficiently than the SN method. The MODFLOW-LGR results were compared with the results obtained using a globally coarse (GC) grid. The LGR simulations required one to two orders of magnitude longer run times than the GC model. However, the improvements of the numerical resolution around the buried valley substantially increased the accuracy of simulated heads and flows compared with the GC simulation. Accuracy further increased locally around the valley flanks when improving the geological resolution using the refined grid. Finally, comparing MODFLOW-LGR simulation with a globally refined (GR) grid showed that the refinement proportion of the model should not exceed 10% to 15% in order to secure method efficiency. PMID- 21623781 TI - Use of organic acids to inactivate Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on organic fresh apples and lettuce. AB - This study was undertaken to investigate the antimicrobial effect of organic acids against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on whole red organic apples and lettuce. Several studies have been conducted to evaluate organic acids as sanitizers. However, no studies have compared antimicrobial effects of various organic acids on organic fresh produce, including evaluation of color changes of produce. Apples and lettuce were inoculated with a cocktail of 3 strains each of 3 foodborne pathogens provided above and treated with 1% and 2% organic acids (propionic, acetic, lactic, malic, and citric acid) for 0, 0.5, 1, 5, and 10 min. With increasing treatment time and acid concentration, organic acid treatments showed significant reduction compared to the control treatment (distilled water), and differences in antimicrobial effects between organic acids were observed. After 10 min of treatment with 1% and 2% organic acids in apples, propionic (0.92 to 2.75 log reduction), acetic (0.52 to 2.78 log reduction), lactic (1.69 to >3.42 log reduction), malic (1.48 to >3.42 log reduction), and citric acid (1.52 to >3.42 log reduction) exhibited significant (P < 0.05) antibacterial effects against 3 foodborne pathogens compared to the control treatment. In lettuce, propionic (0.93 to 1.52 log reduction), acetic (1.13 to 1.74 log reduction), lactic (1.87 to 2.54 log reduction), malic (2.32 to 2.98 log reduction), and citric acid (1.85 to 2.86 log reduction) showed significant (P < 0.05) effects compared to the control treatment. Changes in sample color subjected to organic acids treatment were not significant during storage. PRACTICAL APPLICATION: It is suggested that organic acids have a potential as sanitizers for organic fresh produce. These data may help the organic produce industry provide safe fresh produce for consumers. PMID- 21623782 TI - Supplementation of alkaline phytase (Ds11) in whole-wheat bread reduces phytate content and improves mineral solubility. AB - In this study, alkaline phytase was added to whole-wheat bread and the phytate content and mineral profiles were compared to commercially available acidic phytase. At neutral pH, some phytate (approximately 20%) was degraded by endogenous phytase in wheat flour, while 40% of phytate was hydrolyzed by alkaline phytase DS11 and a 35% reduction was observed with acidic phytase. Most of the enzymatic activity occurred during the proofing stage, and the rate of reaction depended on pH. DS11 phytase effectively degraded the phytate level within a 30 min treatment at pH 7; however, at least 60 min was needed with acidic phytase to achieve the same hydrolysis level. Mineral profiles were also dramatically affected by the phytate reduction. The biggest increase was observed in Fe2+ by the phytase treatment. The Fe2+ content increased 10-fold at pH 7 and 8-fold at pH 5 with alkaline phytase DS11. Alkaline phytase DS11 was shown to be effective at phytate reduction in whole-wheat bread preparation. Additionally, phytate degradation enhanced the mineral availability of bread. PMID- 21623783 TI - Changes in quality, liking, and purchase intent of irradiated fresh-cut spinach during storage. AB - The use of ionizing radiation to enhance microbial safety of fresh spinach at a maximum dose of 4 kGy has been approved by the U.S. Food and Drug Administration (FDA). However, whether spinach can tolerate those high doses of radiation is unclear. Therefore, this study was conducted to investigate the effects of irradiation and storage on quality, liking, and purchase intent of fresh-cut spinach. The oxygen radical absorbance capacity values and total phenolic content were not consistently affected by irradiation. However, the ascorbic acid content of irradiated sample decreased rapidly during storage, resulting in these samples being lower in ascorbic acid content than controls after 7 and 14 d of storage at 4 degrees C. Sensory evaluation by a 50-member panel revealed that purchase intent and ratings for liking of appearance, aroma, texture, flavor, and overall were not affected by irradiation at doses up to 2 kGy. Therefore, irradiation at doses up to 2 kGy may be used to enhance microbial safety without affecting consumer acceptance or overall antioxidant values of irradiated spinach. PMID- 21623784 TI - Optimization of hot water treatment for removing microbial colonies on fresh blueberry surface. AB - Blueberries for the frozen market are washed but this process sometimes is not effective or further contaminates the berries. This study was designed to optimize conditions for hot water treatment (temperature, time, and antimicrobial concentration) to remove biofilm and decrease microbial load on blueberries. Scanning electron microscopy (SEM) image showed a well-developed microbial biofilm on blueberries dipped in room temperature water. The biofilm consisted of yeast and bacterial cells attached to the berry surface in the form of microcolonies, which produced exopolymer substances between or upon the cells. Berry exposure to 75 and 90 degrees C showed little to no microorganisms on the blueberry surface; however, the sensory quality (wax/bloom) of berries at those temperatures was unacceptable. Response surface plots showed that increasing temperature was a significant factor on reduction of aerobic plate counts (APCs) and yeast/mold counts (YMCs) while adding Boxyl(r) did not have significant effect on APC. Overlaid contour plots showed that treatments of 65 to 70 degrees C for 10 to 15 s showed maximum reductions of 1.5 and 2.0 log CFU/g on APCs and YMCs, respectively; with acceptable level of bloom/wax score on fresh blueberries. This study showed that SEM, response surface, and overlaid contour plots proved successful in arriving at optima to reduce microbial counts while maintaining bloom/wax on the surface of the blueberries. PRACTICAL APPLICATION: Since chemical sanitizing treatments such as chlorine showed ineffectiveness to reduce microorganisms loaded on berry surface (Beuchat and others 2001, Sapers 2001), hot water treatment on fresh blueberries could maximize microbial reduction with acceptable quality of fresh blueberries. PMID- 21623785 TI - Effectiveness of edible coatings combined with mild heat shocks on microbial spoilage and sensory quality of fresh cut broccoli (Brassica oleracea L.). AB - The use of edible coatings and mild heat shocks is proposed as postharvest treatments to prevent microbial deterioration of refrigerated broccoli. Minimally processed broccoli was coated with either chitosan or carboxymethyl-cellulose (CMC) combined or not with a previous application of a mild heat shock. The evolution of microbial populations (mesophilic, psycrotrophic, Enterobacteriaceae, molds and yeast, and lactic acid bacteria) was studied during 20 d of storage and fitted to Gompertz and logistic models. Results revealed that, at the end of the storage, chitosan coating significantly reduced all microbiological population counts, except lactic acid bacteria; while higher reduction was observed with chitosan coating combined with a heat shock treatment. A significant delay at the beginning of the exponential phase was observed for all the bacterial populations analyzed. On the other hand, CMC coating, with and without a previous thermal treatment, did not exert any antibacterial effect. Excellent agreement was found between experimental microbial counts and predicted values obtained from Gompertz and logistic models. Kinetic modeling was found to be valuable for prediction of microbiological shelf life of broccoli during storage. Results showed that the application of chitosan coating effectively maintained microbiological quality and extended shelf life of minimally processed broccoli. According to these results, the use of the edible chitosan coating alone or in combination with a heat mild shock appear to be a viable alternative for controlling microbiological growth and sensory attributes in minimally processed broccoli. PRACTICAL APPLICATION: The continuous consumer interest in high quality and food safety, combined with environmental concern has induced to the development and study of edible coatings that avoid the use of synthetic materials. The edible coatings, formed from generally recognized as safe materials, have the potential to reduce weight loss, respiration rate, and improve food appearance and integrity. It is one of the most effective methods to maintain food quality. On the other hand, heat treatments have been demonstrated to be effective as a nonchemical means of improving postharvest quality for a variety of horticultural products. The applications of mild heat shocks combined with edible coatings constitute an alternative for the natural preservation of crops for which the use of synthetic chemicals is objectionable. PMID- 21623786 TI - Pomegranate (Punica granatum) juices: chemical composition, micronutrient cations, and antioxidant capacity. AB - Phenolics, flavonoids, anthocyanins, and tannins of pomegranate juices, obtained from 9 Tunisian ecotypes were quantified. Phenolics and flavonoids in the variety Tounsi (TN) (3299 mg gallic acid equivalents [GAE]/L and 636 mg quercetin equivalents [QE]/L of juice, respectively) were higher than in the variety Gabsi (GB) (1570 mg GAE/L and 135 mg QE/L of juice, respectively). The highest anthocyanins quantity was found in GB 2 with 156 mg cyanidin-3-glucoside equivalents (CGE)/L. TN 3 ecotype showed the highest tannins quantity with 2550 mg catechin equivalents (CE)/L of juice. TN 1 presented the highest radical scavenging activity (2, 2'-azinobis-3-ethylbenzothiazoline-6-sulphonate [ABTS], IC50 [50% inhibition concentration] = 525 mg/L), as well as the highest concentration of micronutrient cations (potassium and sodium). A high correlation (R2 = 0.80) between antioxidant capacity and proanthocyanin contents was found, this suggests that proanthocyanins are the principal contributor in the antioxidant capacity of pomegranate. Our data suggest also that the high concentrations of K+ and Na+ may play a role in the adaptation of pomegranate to arid environments. PMID- 21623787 TI - Growth promotion of Bifidobacterium species by poultry bone and meat trimming hydrolyzate. AB - The growth of bifidobacteria that are employed in the production of functional food is often slow or limited, even on synthetic media. In this study, we investigated whether a peptide hydrolyzate (functional animal protein [FAP]), from poultry bones and meat trimmings, could be a potential source of growth stimulators. The bifidogenic activity of FAP on 18 strains of Bifidobacterium species was assessed via 2 different techniques: turbidimetric measurements and a direct count by fluorescence microscopy. Growth experiments were performed in B12 broth as the basal medium, B12 broth supplemented with N-acetylglucosamine, and B12 broth supplemented with FAP. FAP supplementation yielded the highest maximum optical density (OD) and count values. The use of the microscopic fluorescence counts allowed for better evaluation of the extent of growth and assessment of the viability of cells. FAP from poultry bones and meat trimmings has potential as a growth stimulator for different bifidobacteria of human origin. FAP is a promising ingredient for inclusion in industrial media that are used to culture probiotic strains, including bifidobacteria, because it supports growth very well and maintains cells at a high level of viability. PRACTICAL APPLICATION: Proteinaceous hydrolyzate can be considered a promising ingredient for industrial media that are used to culture probiotic strains, including bifidobacteria, because it improves bacterial growth and maintains cells at a high level of viability. PMID- 21623788 TI - Effects of roasting conditions on the physicochemical properties and volatile distribution in perilla oils (Perilla frutescens var. japonica). AB - Perilla seeds have more than 60% of alpha-linolenic acid, one of omega-3 essential fatty acids. Headspace volatiles and physicochemical properties including color, fluorescence intensity, and the oxidation products in perilla oil (PO) from perilla seeds roasted at different conditions were analyzed. Roasting temperature was 150, 170, 190, and 210 degrees C, and roasting time was 15 and 30 min at each roasting temperature. PO from higher roasting temperature and longer roasting time had lower L* values, higher a*, b*, and chroma values, more brown pigments and fluorescence intensity, and more conjugated dienoic acids. Pyrazines were major volatiles in PO, and furans, sulfur-containing compounds, and hydrocarbons were also detected by a solid phase microextraction gas chromatography/mass spectrometry. In PO, 2,5-Dimethylpyrazine and 2 furancarboxaldehyde were 2 major volatiles. The principal component analysis of volatiles showed the 1st principal component (PC1) and the 2nd principal component (PC2) express 56.64% and 22.72% of the volatile variability in PO, respectively, which can differentiate PO prepared from roasting conditions clearly. Some physicochemical properties especially brown pigment and volatiles were positively correlated with each other in PO. PRACTICAL APPLICATION: Perilla oil (PO) from perilla seeds possesses more than 60% of alpha-linolenic acid, one of omega-3 fatty acids. Roasting process has been used to extract oil from perilla seeds. Understanding physicochemical properties of PO from diverse roasting conditions are important steps to produce PO in food industry. Roasting process induces darkening of color, increase of fluorescence intensity, and brown pigments in PO. Pyrazines and furans are major headspace volatiles in PO roasted above 170 degrees C. The results of this study can help to produce PO in industrial scales with desired headspace volatiles, colors, and oxidative state. PMID- 21623789 TI - Correlating changes that occur in chemical properties with the generation of antioxidant capacity in different sugar-amino acid Maillard reaction models. AB - We investigated the development of antioxidant activity relative to the change of pH, fluorescent intensity, ultraviolet (UV) absorbance (A294), browning (A420), and alpha-dicarbonyl compounds in sugar-amino acid Maillard reaction (MR) model systems comprising fructose, glucose, or ribose each with glycine (Fru-Gly, Glu Gly, and Rib-Gly) or lysine (Fru-Lys, Glu-Lys, and Rib-Lys), respectively, which were heated at 121 degrees C for 5 to 90 min. For hexose models, the change in pH was shown to fit a second-order polynomial regression with A294 and A420. Antioxidant activity was significantly and positively correlated with UV absorbance (r = 0.905, P < 0.001) and browning products (r = 0.893, P < 0.001) rather than with fluorescent products or the alpha-dicarbonyl compounds. Type of sugar was most important in evoking a change in UV absorbance, browning, alpha dicarbonyl compounds, and antioxidant activity of MR products (MRPs). In conclusion, the antioxidant activity of MRPs in six model systems was more closely associated with products derived at the intermediate-to-late stages of the reaction and influenced mostly by the type of sugar. PRACTICAL APPLICATION: We report on the different factors and their interactions that are important for understanding the functional attributes of food components that comprise the generation of Maillard browning products and the associated antioxidant activities generated during high-temperature food processing. PMID- 21623790 TI - Effect of irradiation on Salmonella survival and quality of 2 varieties of whole green onions. AB - Two varieties of green onions, Banner and Baja Verde, were inoculated with a cocktail of 3 Salmonella strains using dip and spot inoculation and irradiated at 0, 0.3, 0.6, 0.9, and 1.2 kGy using electron beam. Salmonella survivors were enumerated using a XLD underlay/TSAYE overlay plating method. The D values were in the range of 0.26 to 0.32 kGy depending on variety but not on the method of inoculation. This indicated that a 5-log reduction of Salmonella can be achieved at a dose of 1.6 kGy. For the quality study, both varieties of green onions were irradiated at 0, 1.5, 2.0, and 2.5 kGy and evaluated for changes in microbial counts, color, texture, and visual quality during storage at 4 degrees C. Irradiation reduced total plate counts and psychrotrophs by 3 logs. Although the counts increased during storage, they did not exceed the initial counts of control. No significant difference was observed in color and texture between irradiated samples and control. The control maintained good visual quality for about 13 d as compared to 15 d for 1.5 and 2.5 kGy samples. The 2.0 kGy samples maintained good visual quality for 17 d suggesting that irradiation can increase shelf life by reducing spoilage microorganisms but higher doses can be detrimental to quality. At the dose levels required to achieve a 5-log reduction in Salmonella, the shelf life of whole green onion can be extended. This study shows that irradiation can be used to enhance safety without adverse effects on quality. PMID- 21623791 TI - Highlights of the optical highlighter fluorescent proteins. AB - The development of super-resolution microscopy techniques using molecular localization, such as photoactivated localization microscopy, fluorescence photoactivated localization microscopy, stochastic optical reconstruction microscopy, photoactivated localization microscopy with independent running acquisition and many others, has heightened interest in molecules that will be grouped here into a category referred to as 'optical highlighter' fluorescent proteins. This review will survey many of the advances in development of fluorescent proteins for optically highlighting sub-populations of fluorescently labelled molecules. PMID- 21623792 TI - Pituitary adenylate cyclase-activating peptide induces long-lasting neuroprotection through the induction of activity-dependent signaling via the cyclic AMP response element-binding protein-regulated transcription co-activator 1. AB - Pituitary adenylate cyclase-activating peptide (PACAP) is a neuroprotective peptide which exerts its effects mainly through the cAMP-protein kinase A (PKA) pathway. Here, we show that in cortical neurons, PACAP-induced PKA signaling exerts a major part of its neuroprotective effects indirectly, by triggering action potential (AP) firing. Treatment of cortical neurons with PACAP induces a rapid and sustained PKA-dependent increase in AP firing and associated intracellular Ca(2+) transients, which are essential for the anti-apoptotic actions of PACAP. Transient exposure to PACAP induces long-lasting neuroprotection in the face of apoptotic insults which is reliant on AP firing and the activation of cAMP response element (CRE) binding protein (CREB)-mediated gene expression. Although direct, activity-independent PKA signaling is sufficient to trigger phosphorylation on CREB's activating serine-133 site, this is insufficient for activation of CREB-mediated gene expression. Full activation is dependent on CREB-regulated transcription co-activator 1 (CRTC1), whose PACAP induced nuclear import is dependent on firing activity-dependent calcineurin signaling. Over-expression of CRTC1 is sufficient to rescue PACAP-induced CRE mediated gene expression in the face of activity-blockade, while dominant negative CRTC1 interferes with PACAP-induced, CREB-mediated neuroprotection. Thus, the enhancement of AP firing may play a significant role in the neuroprotective actions of PACAP and other adenylate cyclase-coupled ligands. PMID- 21623793 TI - AMP-activated protein kinase: a potential player in Alzheimer's disease. AB - AMP-activated protein kinase (AMPK) stimulates energy production via glucose and lipid metabolism, whereas it inhibits energy consuming functions, such as protein and cholesterol synthesis. Increased cytoplasmic AMP and Ca(2+) levels are the major activators of neuronal AMPK signaling. Interestingly, Alzheimer's disease (AD) is associated with several abnormalities in neuronal energy metabolism, for example, decline in glucose uptake, mitochondrial dysfunctions and defects in cholesterol metabolism, and in addition, with problems in maintaining Ca(2+) homeostasis. Epidemiological studies have also revealed that many metabolic and cardiovascular diseases are risk factors for cognitive impairment and sporadic AD. Emerging studies indicate that AMPK signaling can regulate tau protein phosphorylation and amyloidogenesis, the major hallmarks of AD. AMPK is also a potent activator of autophagic degradation which seems to be suppressed in AD. All these observations imply that AMPK is involved in the pathogenesis of AD. However, the responses of AMPK activation are dependent on stimulation and the extent of activating stress. Evidently, AMPK signaling can repress and delay the appearance of AD pathology but later on, with increasing neuronal stress, it can trigger detrimental effects that augment AD pathogenesis. We will outline the potential role of AMPK function in respect to various aspects affecting AD pathogenesis. PMID- 21623795 TI - Up-regulation of the mitochondrial malate dehydrogenase by oxidative stress is mediated by miR-743a. AB - These experiments reveal for the first time that microRNAs (miRNAs) mediate oxidant regulated expression of a mitochondrial tricarboxylic acid cycle gene (mdh2). mdh2 encoded malate dehydrogenase (MDH) is elevated by an unknown mechanism in brains of patients that died with Alzheimer's disease. Oxidative stress, an early and pervasive event in Alzheimer's disease, increased MDH activity and mRNA level of mdh2 by 19% and 22%, respectively, in a mouse hippocampal cell line (HT22). Post-transcriptional events underlie the change in mRNA because actinomycin D did not block the elevated mdh2 mRNA. Since miRNAs regulate gene expression post-transcriptionally, the expression of miR-743a, a miRNA predicted to target mdh2, was determined and showed a 52% reduction after oxidant treatment. Direct interaction of miR-743a with mdh2 was demonstrated with a luciferase based assay. Over-expression or inhibition of miR-743a led to a respective reduction or increase in endogenous mRNA and MDH activity. The results demonstrate that miR-743a negatively regulates mdh2 at post-transcriptional level by directly targeting the mdh2 3'UTR. The findings are consistent with the suggestion that oxidative stress can elevate the activity of MDH through miR 743a, and provide new insights into possible roles of miRNA in oxidative stress and neurodegeneration. PMID- 21623796 TI - Exploiting the Brachypodium Tool Box in cereal and grass research. AB - It is now a decade since Brachypodium distachyon (Brachypodium) was suggested as a model species for temperate grasses and cereals. Since then transformation protocols, large expressed sequence tag (EST) databases, tools for forward and reverse genetic screens, highly refined cytogenetic probes, germplasm collections and, recently, a complete genome sequence have been generated. In this review, we will describe the current status of the Brachypodium Tool Box and how it is beginning to be applied to study a range of biological traits. Further, as genomic analysis of larger cereals and forage grasses genomes are becoming easier, we will re-evaluate Brachypodium as a model species. We suggest that there remains an urgent need to employ reverse genetic and functional genomic approaches to identify the functionality of key genetic elements, which could be employed subsequently in plant breeding programmes; and a requirement for a Pooideae reference genome to aid assembling large pooid genomes. Brachypodium is an ideal system for functional genomic studies, because of its easy growth requirements, small physical stature, and rapid life cycle, coupled with the resources offered by the Brachypodium Tool Box. PMID- 21623794 TI - Ascl1-induced neuronal differentiation of P19 cells requires expression of a specific inhibitor protein of cyclic AMP-dependent protein kinase. AB - cAMP-dependent protein kinase (PKA) plays a critical role in nervous system development by modulating sonic hedgehog and bone morphogenetic protein signaling. In the current studies, P19 embryonic carcinoma cells were neuronally differentiated by expression of the proneural basic helix-loop-helix transcription factor Ascl1. After expression of Ascl1, but prior to expression of neuronal markers such as microtubule associated protein 2 and neuronal beta tubulin, P19 cells demonstrated a large, transient increase in both mRNA and protein for the endogenous protein kinase inhibitor (PKI)beta. PKIbeta-targeted shRNA constructs both reduced the levels of PKIbeta expression and blocked the neuronal differentiation of P19 cells. This inhibition of differentiation was rescued by transfection of a shRNA-resistant expression vector for the PKIbeta protein, and this rescue required the PKA-specific inhibitory sequence of the PKIbeta protein. PKIbeta played a very specific role in the Ascl1-mediated differentiation process as other PKI isoforms were unable to rescue the deficit conferred by shRNA-mediated knockdown of PKIbeta. Our results define a novel requirement for PKIbeta and its inhibition of PKA during neuronal differentiation of P19 cells. PMID- 21623797 TI - Photocatalytic reduction of CO2 over Cu-TiO2 /molecular sieve 5A composite. AB - TiO(2) and different Cu wt% loaded TiO(2) (TC(0.5-5.0)), 10 wt% TC(2.0) supported on molecular sieve 5A (10 wt% TC(2.0)/MS) were prepared by impregnation and solid state dispersion methods. The photocatalysts prepared were characterized using XRD, SEM, and UV-Vis DRS, TEM, XPS spectroscopy techniques. Photocatalytic reduction of CO(2) in water and alkaline solution are investigated in a batch reactor. The yield of oxalic acid increased notably when TC was supported on molecular sieve. The Cu-TiO(2) supported on molecular sieve catalyst promotes the charge separation that leads to an increase in the selective formation of oxalic acid in addition to methanol, acetic acid and traces of methane. The product formation is due to the high adsorption of CO(2), water and product shape selectivity of the composite photocatalyst. The maximum yield of oxalic acid was found to be 65.6 MUg h(-1) g(-1) per cat using 0.2 N NaOH containing solution over 10 wt% TC(2.0)/MS photocatalyst. The difference in the photocatalytic activity is related to its physicochemical properties. PMID- 21623798 TI - Anion/cation induced optical switches based on luminescent lanthanide (Tb3+ and Eu3+) hydrogels. AB - Two novel silica based lanthanide complexes (Tb(a)(2) and Eu(a)(2)) were encapsulated into poly(acrylic acid) host. Both Tb(III) and Eu(III) containing hydrogels have typical and easily distinguished narrow line emissions occurring in the green and red region respectively. Particularly, the excitation wavelength for Eu complex can be extended into nearly visible light range (lambda(ex) = 395 nm). Interestingly, we discover that these target materials not only exhibit selective emission response towards HSO(4)(-) (detection limit 10(-5) M) compared with CH(3)COO(-), F(-), Cl(-), Br(-) and I(-) but also give unique quenching to Cu(2+) (detection limit 10(-5) M) (tested cations: Cu(2+), Pd(2+), Cd(2+), Co(2+) and Mn(2+)). More importantly, this kind of materials can be recycled more than 10 times. PMID- 21623799 TI - Stomatal factors and vulnerability of stem xylem to cavitation in poplars. AB - The relationships between the vulnerability of stem xylem to cavitation, stomatal conductance, stomatal density, and leaf and stem water potential were examined in six hybrid poplar (P38P38, Walker, Okanese, Northwest, Assiniboine and Berlin) and balsam poplar (Populus balsamifera) clones. Stem xylem cavitation resistance was examined with the Cavitron technique in well-watered plants grown in the greenhouse. To investigate stomatal responses to drought, plants were subjected to drought stress by withholding watering for 5 (mild drought) and 7 (severe drought) days and to stress recovery by rewatering severely stressed plants for 30 min and 2 days. The clones varied in stomatal sensitivity to drought and vulnerability to stem xylem cavitation. P38P38 reduced stomatal conductance in response to mild stress while the balsam poplar clone maintained high leaf stomatal conductance under more severe drought stress conditions. Differences between the severely stressed clones were also observed in leaf water potentials with no or relatively small decreases in Assiniboine, P38P38, Okanese and Walker. Vulnerability to drought-induced stem xylem embolism revealed that balsam poplar and Northwest clones reached loss of conductivity at lower stem water potentials compared with the remaining clones. There was a strong link between stem xylem resistance to cavitation and stomatal responsiveness to drought stress in balsam poplar and P38P38. However, the differences in stomatal responsiveness to mild drought suggest that other drought-resistant strategies may also play a key role in some clones of poplars exposed to drought stress. PMID- 21623800 TI - Continuing professional dental education gains momentum. PMID- 21623801 TI - Applied anatomy of the pterygomandibular space: improving the success of inferior alveolar nerve blocks. AB - A thorough knowledge of the anatomy of the pterygomandibular space is essential for the successful administration of the inferior alveolar nerve block. In addition to the inferior alveolar and lingual nerves, other structures in this space are of particular significance for local anaesthesia, including the inferior alveolar vessels, the sphenomandibular ligament and the interpterygoid fascia. These structures can all potentially have an impact on the effectiveness of local anaesthesia in this area. Greater understanding of the nature and extent of variation in intraoral landmarks and underlying structures should lead to improved success rates, and provide safer and more effective anaesthesia. The direct technique for the inferior alveolar nerve block is used frequently by most clinicians in Australia and this review evaluates its anatomical rationale and provides possible explanations for anaesthetic failures. PMID- 21623802 TI - Contemporary fluid intake and dental caries in Australian children. AB - In Australia, caries experience of 6-year-old and 12-year-old children has increased since the mid to late 1990s. Previously, caries rates had declined, attributable to community water fluoridation. The recent caries increase has been attributed speculatively to changes in fluid intake, including increased consumption of sweet drinks and bottled waters. Increasing urbanization and globalization have altered children's diets worldwide, promoting availability and access to processed foods and sweet drinks. Studies in Australia and internationally have demonstrated significant associations between sweet drink intake and caries experience. Despite widespread fluoride availability in contemporary Australian society, the relationship between sugar consumption and caries development continues and restricting sugar intake remains key to caries prevention. Caries risk assessment should be included in treatment planning for all children; parents should be advised of their child's risk level and given information on oral health promotion. Readily-implemented caries risk assessment tools applicable to parents and clinicians are now available. Public health information should increase awareness that consuming sweet drinks can have deleterious effects on the dentition as well as the potential for promoting systemic disease. Restricting sales of sweet drinks and sweet foods and providing healthy food and drinks for purchase in schools is paramount. PMID- 21623803 TI - Influence of implant abutment angulations on the fracture resistance of overlaying CAM-milled zirconia single crowns. AB - BACKGROUND: An in vitro study was performed to assess the effect of three implant abutment angulations and three core thicknesses on the fracture resistance of overlaying computer-aided manufacturing (CAM) milled zirconia (Cercon((r)) system) single crowns. METHODS: Three groups, coded A to C, with different implant abutment angulations (group A/0 degrees , group B/15 degrees and group C/30 degrees angulation) were used to construct 15 crowns for each angulation. Forty-five overlay restorations were milled using the Cercon((r)) system with zirconium core thicknesses of 0.4, 0.6 and 0.8 mm using five crowns for each angulation. The final restorations were prepared and stored in distilled water at mouth temperature (37 degrees C) for 24 hours prior to testing. The restorations were cemented using Temp Bond((r)) . The load required to break each crown and the mode of failure were recorded. All the results obtained were statistically analysed by the ANOVA test (level of significance p < 0.05). Tested crowns were examined using a stereomicroscope at 40X and selected crowns (five randomly selected from each group were further examined by scanning electron microscopy) to reveal the zirconia-ceramic interface and to determine the fracture origin. RESULTS: Implant abutment angulations significantly (p < 0.05) reduced the fracture resistance of overlaying CAM-milled zirconia single crowns. The fracture loads of Cercon((r)) crowns cemented onto abutment preparations with a 30 degrees angulation were the lowest of the groups tested. The core thickness (0.4 to 0.8 mm) did not significantly (p > 0.05) affect the fracture resistance of the CAM milled zirconia single crowns. SEM showed that the origin of the fracture appeared to be located at the occlusal surfaces of the crowns and the crack propagation tended to radiate from the occlusal surface towards the gingival margin. CONCLUSIONS: The implant angulation of 30 degrees significantly (p < 0.05) reduced the fracture resistance of overlaying CAM-milled zirconia single crowns. Reducing the core thickness from 0.8 mm to 0.4 mm did not affect (p > 0.05) the fracture resistance of overlaying CAM-milled zirconia single crowns. PMID- 21623804 TI - Augmentation of linear facial anthropometrics through modern morphometrics: a facial convexity example. AB - BACKGROUND: The facial region has traditionally been quantified using linear anthropometrics. These are well established in dentistry, but require expertise to be used effectively. The aim of this study was to augment the utility of linear anthropometrics by applying them in conjunction with modern 3-D morphometrics. METHODS: Facial images of 75 males and 94 females aged 18-25 years with self-reported Caucasian ancestry were used. An anthropometric mask was applied to establish corresponding quasi-landmarks on the images in the dataset. A statistical face-space, encoding shape covariation, was established. The facial median plane was extracted facilitating both manual and automated indication of commonly used midline landmarks. From both indications, facial convexity angles were calculated and compared. The angles were related to the face-space using a regression based pathway enabling the visualization of facial form associated with convexity variation. RESULTS: Good agreement between the manual and automated angles was found (Pearson correlation: 0.9478-0.9474, Dahlberg root mean squared error: 1.15 degrees -1.24 degrees ). The population mean angle was 166.59 degrees -166.29 degrees (SD 5.09 degrees -5.2 degrees ) for males females. The angle-pathway provided valuable feedback. CONCLUSIONS: Linear facial anthropometrics can be extended when used in combination with a face-space derived from 3-D scans and the exploration of property pathways inferred in a statistically verifiable way. PMID- 21623805 TI - Severe adverse reactions to dental local anaesthetics: systemic reactions. AB - BACKGROUND: Occasionally, patients suffer systemic adverse effects from injections of local anaesthetic solutions. This may range from minor transient vasovagal attacks to life-threatening collapse. METHODS: The suspected adverse reactions reported to the Office of Product Review of the Therapeutic Goods Administration (TGA) were analysed in detail. RESULTS: There was a high incidence (70%) of adverse reactions associated with prilocaine, which is much greater than its market share (less than 20%). There is a tendency to consider all systemic adverse reactions as being 'allergic' reactions although this is rarely the case. Syncope, cardiovascular and central nervous system reactions are much more common. There is also a risk of methaemoglobinaemia to prilocaine and articaine. A small series of cases referred to one of the authors were also reported. CONCLUSIONS: Recommendations are made as to the prevention, acute care and subsequent investigation of adverse reactions. The most important conclusion is not to just label the response as allergic and to use an alternative agent. Detailed investigation and reporting should be made for all cases of suspected severe adverse reaction to local anaesthetic agents. PMID- 21623806 TI - Severe adverse reactions to dental local anaesthetics: prolonged mandibular and lingual nerve anaesthesia. AB - BACKGROUND: Prolonged anaesthesia may occur following dental local anaesthetic blocks. This paper reviews the possible mechanisms of injury. Direct injury to the nerve by the needle, although commonly thought to be the mechanism, is unlikely. It is much more likely that the injury is from neurotoxicity and/or interference with the vascularization of the nerve. METHODS: Estimation of the frequency of injury was complicated by the fact that although local anaesthetics are prescription-only (S4) drugs, they are supplied without prescription by dental supply houses. Unlike all other S4 drugs, there is no statutory requirement to record supply. The pharmaceutical and supply houses relied on that and 'commercial confidentiality' to not supply information. RESULTS: An informed estimate of 1 in 27 415 was made but this figure has wide confidence limits. Management of cases of prolonged anaesthesia following local anaesthetic injection is discussed. CONCLUSIONS: Patients who suffer this uncommon complication suffer considerable distress and feel injured, so care must be exhibited in their management. Specialist referral is recommended. PMID- 21623807 TI - Oral rehabilitation with dental implants after cancer treatment. AB - BACKGROUND: Patients who undergo surgical management of oral cancer may greatly benefit from an implant-supported prosthesis. This study reports on the clinical experience of dental implant placement in patients following resection of oral cancer over a 15-year period. Controversies including the use of dental implants in irradiated tissues, and hyperbaric oxygen treatment will also be discussed. METHODS: Thirty-one patients who had dental implants placed as part of their oral rehabilitation between 1992 and 2007 were investigated. Demographic data and factors including implant survival, type of prosthesis provided, radiotherapy and the hyperbaric oxygen therapy were analysed. RESULTS: In this retrospective study, there was a retention rate of 110 implants from a total of 115 implants placed. A high rate of implant retention was found, with 5 implant failures from a total of 115 implants placed. The 5 failed implants occurred in free flap bone that had been irradiated. CONCLUSIONS: Dental implants provide an important role in the oral rehabilitation of oral cancer patients. There may be an increased risk of implant failure in free flap bone that has been irradiated. PMID- 21623808 TI - Mandible fracture severity may be increased by alcohol and interpersonal violence. AB - BACKGROUND: Alcohol as a cofactor in interpersonal violence (IPV) has been established by studies from a number of countries. This study aimed to determine if alcohol was a cofactor in the incidence or severity of mandible fracture. METHODS: A prospective study of mandible fracture patients presenting for oral maxillofacial review over 16 months was completed. Injury severity was assessed utilizing the Mandible Injury Severity Score (MISS). RESULTS: A total of 252 facial trauma cases presented to our tertiary referral centre, 83 with fractures of the mandible. The majority of presentations were secondary to IPV (n = 54, 65.06%), 49 (90.74%) of these cases involved alcohol. Overall, alcohol was involved in 63.85% of cases (n = 53). The relative risk of requiring surgical intervention with alcohol involvement was 2.68 (CI = 1.11-9.47). Alcohol significantly increased facial fracture severity for MISS: alcohol (n = 53) 13.07 +/- 5.01, no alcohol (n = 30) 11.03 +/- 4.87 (p < 0.05). IPV also increased facial fracture severity for MISS: IPV (n = 54) 13.09 +/- 4.90, non-IPV (n = 29) 11.00 +/- 4.81 (p < 0.05). The angle of the mandible was most commonly fractured (40.5% of cases). CONCLUSIONS: Mandible fracture patients, whose injury is a result of IPV, have more severe fractures and a higher likelihood of requiring surgery if alcohol is involved. This correlates to a higher surgical workload, economic and social burden to the community. Primary alcohol and IPV prevention strategies will play an important role in reducing mandible fracture. PMID- 21623809 TI - Relationship of unstimulated saliva cortisol level with severity of oral dryness feeling in menopausal women. AB - BACKGROUND: Menopause may be associated with some adverse changes, such as oral dryness (OD) feeling. The exact mechanisms that mediate sensation of OD in menopausal women have not been firmly established. The purpose of this study was to assess the relationship of unstimulated whole saliva cortisol level with severity of OD feeling in menopausal women. METHODS: A case control study was carried out on 70 selected menopausal women with/without OD feeling, conducted at the Clinic of Oral Medicine, Tehran University of Medical Sciences, Iran. The xerostomia inventory (XI) score was used as an index of OD feeling severity. Unstimulated whole saliva cortisol concentration was measured by ELISA. Statistical analysis of student's t-test and Spearman correlation was used. RESULTS: The mean cortisol concentration of saliva was significantly higher in women with OD feeling than the control. There was significant positive correlation between the XI score and the concentration of unstimulated whole saliva cortisol (r = 0.559, p = 0.025). CONCLUSIONS: Unstimulated whole saliva cortisol is higher in menopausal women with OD feeling than in controls and this correlates with the severity of OD. PMID- 21623810 TI - Tricalcium silicate induced mineralization for occlusion of dentinal tubules. AB - BACKGROUND: The aim of this study was to evaluate the dentinal tubule occluding ability of tricalcium silicate (Ca(3) SiO(5) or C(3) S) in vitro. METHODS: Dentine discs were prepared from extracted caries-free human third molars and etched with 0.02 M citric acid (pH 4) for 3 minutes in order to produce patent dentinal tubules. Tricalcium silicate pastes of different viscosities were applied to the surface of the dentine specimens for 3 minutes and then removed with a swab followed by deionized water rinse. The dentine specimens were immersed in artificial saliva for 7 days. The tubule occlusion, sealing depth and chemical composition of the dentine specimens and bonding force between the dentine matrix and mineralized layer were evaluated by scanning electron microscopy, energy dispersion X-ray spectroscopy, X-ray diffraction analysis and a nanoscratch test. RESULTS: A homogeneous layer of mineral crystals was precipitated onto the dentine surface and caused significant occlusion of the dentinal tubules. The dentine permeability and sealing depth of the C(3) S increased with the decrease of viscosity of the paste. With a proper viscosity of C(3) S pastes, the interface between the mineralized layer and the dentine matrix bonded well. CONCLUSIONS: This study suggests that C(3) S may be an effective agent for the treatment of dentine hypersensitivity. PMID- 21623811 TI - Prosthodontic considerations designed to optimize outcomes for single-tooth implants. A review of the literature. AB - BACKGROUND: The aim of this study was to review the literature on the restoration of single-tooth implants, and to develop evidence-based conclusions to optimize aesthetic, biologic and patient-related outcomes. METHODS: An electronic and hand search was conducted using the search terms 'dental implants, single-tooth; dental restoration, temporary; dental impression materials; dental impression technique; dental prosthesis, implant-supported; dental prosthesis design; dental abutments; dental occlusion; maintenance; survival; and survival analysis'. Resultant titles were screened, and full text was obtained where relevant. The authors selected the most appropriate articles, giving preference to systematic reviews and long-term, patient-based outcome data. RESULTS: Thirty-nine articles were selected and critiqued by the authors. CONCLUSIONS: There was strong suggestion by several authors that peri-implant soft tissue aesthetics can be sculpted through provisional restoration contour, but there are no clinical outcome studies to define or support this claim. Laboratory studies demonstrate that pick-up type impression copings in conjunction with elastomeric impressions are the most accurate means for transferring implant position to a dental cast. Laboratory and finite-element analysis studies suggest implants with an internal type connection show improved stress distribution, but supportive clinical data are lacking. The authors of this review favour a screw-retained prosthesis for retrievability. Clinical and histological studies show that gold, titanium and zirconia ceramic abutment materials exhibit excellent biological responses, although there is insufficient data on the clinical service provided by zirconia as an implant-substructure material. The literature does not associate any particular occlusal scheme with superior clinical outcomes. Implant-borne single crowns offer comparable clinical service to tooth-borne fixed dental prostheses. However, single-tooth implant restorations are associated with an increased incidence of biological and technical complications. PMID- 21623812 TI - Prevention of enamel demineralization after tooth bleaching by bioactive glass incorporated into toothpaste. AB - BACKGROUND: The aim of this study was to determine the effects of bleaching on the structure of the enamel layer of teeth and the potential of the commercial bioactive glass NovaMin(r) in two different toothpastes to remineralize such regions of the enamel. Three aspects were considered: the extent and nature of the alterations in the enamel after application of the bleaching agents; the extent of remineralization after application of two commercial toothpastes containing bioactive glass; and whether or not there were differences between the toothpastes in terms of their effectiveness in promoting remineralization. METHODS: Bleaching agent based on 16% carbamide peroxide was applied to the enamel surface of freshly extracted human molars for 8 minutes, once a day for 7 days. After the bleaching cycles, the enamel surface was analysed by SEM and EDX. RESULTS: The results obtained in the study lead to the conclusion that application of 16% carbamide peroxide causes distinct morphological changes to the enamel surface which vary from mild to severe. Subsequent treatment with either of the toothpastes containing the bioactive glass NovaMin(r) resulted in the formation of a protective layer on the enamel surface, consisting of bioactive glass deposits, with only slight differences between the two brands. Application of these dentifrices also caused increases in the Ca and P content of the enamel layer, returning it to that of undamaged enamel. CONCLUSIONS: Remineralizing toothpastes should be used after bleaching, in order to repair any damage to the mineral tissue caused by these procedures. PMID- 21623813 TI - The evaluation of bacterial flora in progress of peri-implant disease. AB - BACKGROUND: Cumulative interceptive supportive therapy (CIST) is currently used as a guideline for treating peri-implant diseases. The objectives of this study were to determine the detection rate and measure the number of periodontopathic bacteria in lesions of different CIST levels and thereby characterize peri implant disease from a bacteriological viewpoint. METHODS: This study included 105 patients who had both residual natural teeth and implants with peri-implant disease. A total of 105 implants were divided into levels A, B, C and D according to the CIST classification. Bacterial samples were collected from peri-implant pockets and four periodontopathic bacteria were measured by PCR and PCR-Invader assay. RESULTS: The number of periodontopathic bacteria increased in line with CIST level, and the detection rate was also associated with CIST level. However, no difference was found in the bacterial detection rate of P. gingivalis and T. denticola between CIST-B and CIST-C. There was a higher detection rate of all periodontopathic bacteria for CIST-D. CONCLUSIONS: The number of periodontopathic bacteria and detection rate increased as peri-implant disease advanced. However, there were no major differences in the detection rate between CIST-B and CIST-C. On the other hand, a higher detection rate of periodontopathic bacteria was seen for CIST-D. PMID- 21623814 TI - Management of intrusive luxation in the primary dentition by surgical repositioning: an alternative approach. AB - Intrusive luxation is the most common trauma during early infancy which results in the displacement of the tooth into its alveolus. Depending on the severity of the intrusion, the American Academy of Pediatric Dentistry recommends either extraction or spontaneous re-eruption for the primary tooth. This case report provides a brief insight into surgical repositioning as an alternative treatment option for the management of intruded primary tooth in a 4-year-old girl. PMID- 21623815 TI - Maxillary tuberosity fracture: a life-threatening haemorrhage following simple exodontia. AB - Maxillary tuberosity fracture is a potential complication of routine exodontia of posterior maxillary teeth. On rare occasions, such a complication can result in torrential haemorrhage due to the close proximity of significant vessels to the area. We present a case of life-threatening haemorrhage complicating a tuberosity fracture during simple extraction of a maxillary posterior tooth. The local anatomy of the region is discussed and we provide guidelines for general dental practitioners for the management of the complication of a tuberosity fracture during routine exodontia. PMID- 21623816 TI - Tuberculosis cutis orificialis with both gingival involvement and underlying pulmonary tuberculosis. AB - Tuberculosis cutis orificialis is a rare manifestation of cutaneous tuberculosis which occurs in the oral, perianal and adjacent mucosa. The most frequent orificial lesion location is the tongue. We report a case of a 35-year-old female with tuberculosis cutis orificialis with gingival involvement. She had a six month history of a moderately painful gingival ulceration. Histopathological examination showed granulomatous infiltrates composed of epithelioid cells, Langhans giant cells and caseating necrosis. A purified protein derivative test was positive. Sputum culture was positive for Mycobacterium tuberculosis. Chest X ray and high resolution computed tomography showed active pulmonary tuberculosis in both upper lung zones. The gingival specimen was positive for M. tuberculosis polymerase chain reaction. A complete resolution was achieved after six months of anti-tuberculosis therapy. Dental identification of M. tuberculosis may serve as an important aid in the first line of control of this dangerous infectious disease. PMID- 21623817 TI - Consensus statement by hospital based dentists providing dental treatment for patients with inherited bleeding disorders. AB - Avoidance of dental care and neglect of oral health may occur in patients with inherited bleeding disorders because of concerns about perioperative and postoperative bleeding, but this is likely to result in the need for crisis care, and more complex and high-risk procedures. Most routine dental care in this special needs group can be safely managed in the general dental setting following consultation with the patient's haematologist and adherence to simple protocols. Many of the current protocols for dental treatment of patients with inherited bleeding disorders were devised many years ago and now need revision. There is increasing evidence that the amount of factor cover previously recommended for dental procedures can now be safely reduced or may no longer be required in many cases. There is still a need for close cooperation and discussion between the patient's haematologist and dental surgeon before any invasive treatment is performed. A group of hospital based dentists from centres where patients with inherited bleeding disorders are treated met and, after discussions, a management protocol for dental treatment was formulated. PMID- 21623822 TI - Nursing home care. PMID- 21623823 TI - Communication and dentistry - an important part of the dentist-patient relationship. PMID- 21623824 TI - Dental graduates and student debt. PMID- 21623826 TI - Recurrent oral ulceration. PMID- 21623828 TI - IgE-binding epitopes: a reappraisal. AB - Here, we discuss various questions related to IgE epitopes: What are the technical possibilities and pitfalls, what is currently known, how can we put this information into hypothetical frameworks and the unavoidable question: how useful is this information for patient care or allergenicity prediction? We discuss the information obtained by (i) 3D structures of allergen-antibody complexes; (ii) analysis of allergen analogues; (iii) mimics without obvious structural similarity; (iv) mAbs competing with IgE; (v) repertoire analysis of cloned IgEs, and other developments. Based on limited data, four suggestions are presented in the literature: (i) IgE might be more cross-reactive than IgG; (ii) IgE might be more often directed to immunologically 'uninviting' surfaces; (iii) IgE epitopes may tend to cluster and (iv) IgE paratopes might have a higher intrinsic flexibility. While these are not proven facts, they still can generate hypotheses for future research. The hypothesis is put forward that the IgE repertoire of switched B-cells is less influenced by positive selection, because positive selection might not be able to rescue IgE-switched B cells. While this might be of interest for the discussion about mechanisms leading to allergen sensitization, we need to be modest in answering the 'clinical relevance' question. Current evidence indicates the IgE-epitope repertoire is too big to make specific IgE epitopes a realistic target for diagnosis, treatment or allergenicity prediction. In-depth analysis of a few selected IgE epitope peptides or mimitopes derived from allergen-sequences and from random peptide libraries, respectively, might well prove rewarding in relation to diagnosis and prognosis of allergy, particularly food allergy. PMID- 21623829 TI - A single nucleotide deletion at the C1 inhibitor gene as the cause of hereditary angioedema: insights from a Brazilian family. AB - BACKGROUND: Hereditary angioedema is an autosomal dominant disease characterized by episodes of subcutaneous and submucosal edema. It is caused by deficiency of the C1 inhibitor protein, leading to elevated levels of bradykinin. More than 200 mutations in C1 inhibitor gene have been reported. The aim of this study was to analyze clinical features of a large family with an index case of hereditary angioedema and to determine the disease-causing mutation in this family. METHODS: Family pedigree was constructed with 275 individuals distributed in five generations. One hundred and sixty-five subjects were interviewed and investigated for mutation at the C1 inhibitor gene. Subjects reporting a history of recurrent episodes of angioedema and/or abdominal pain attacks underwent evaluation for hereditary angioedema. RESULTS: We have identified a novel mutation at the C1 inhibitor gene, c.351delC, which is a single-nucleotide deletion of a cytosine on exon 3, resulting in frameshift with premature stop codon. Sequencing analysis of the hypothetical truncated C1 inhibitor protein allowed us to conclude that, if transcription occurs, this protein has no biological activity. Twenty-eight members of the family fulfilled diagnostic criteria for hereditary angioedema and all of them presented the c.351delC mutation. Variation in clinical presentation and severity of disease was observed among these patients. One hundred and thirty-seven subjects without hereditary angioedema did not have the c.351delC mutation. CONCLUSION: The present study provides definitive evidence to link a novel genetic mutation to the development of hereditary angioedema in patients from a Brazilian family. PMID- 21623830 TI - How the human spermatozoa sense the oocyte: a new role of SDF1-CXCR4 signalling. AB - For fertilization to occur in mammals, ejaculated spermatozoa must reach the egg which, following ovulation has moved from the ovary into the Fallopian tube. Two active mechanisms of spermatozoa guidance have been shown in mammals: thermotaxis and chemotaxis. The identity of most of human spermatozoa chemoattractants is unknown, and herein we tested if SDF1 (chemokine stromal cell-derived factor-1) and its pathway is involved in spermatozoa chemotaxis. We found that SDF1 is expressed in the oocytes, endometrium and follicular fluid, as well as its specific receptor CXCR4 (chemokine CXC motif receptor 4) is expressed in the head of spermatozoa. By SDF1 gradient experiments, we stated that SDF1 is able to induce hyperactivation in spermatozoa leading to accumulation, to give rise to an increase in intracellular calcium concentration, and to preserve the mitochondrial status and not to induce acrosome reaction. Our findings suggest these phenomena could reflect spermatozoa chemotaxis, and that SDF1 action could represent an important event leading to egg fertilization, even if further studies regarding the link between spermatozoa accumulation and chemotaxis are mandatory. These data suggest that the SDF-1/CXCR4 signalling could be used to manipulate the human fertilization, to improve both the outcome of physiological or assisted reproduction, and to develop new contraceptive methods, by development of SDF1 or CXCR4 antagonist. PMID- 21623831 TI - Genome-wide association studies provide new insights into the genetic basis of testicular germ-cell tumour. AB - Testicular germ-cell tumour (TGCT) is the most common cancer in young men, and genetic epidemiological studies suggest that the disease has a strong genetic basis. Until 2009, very little of this genetic component had been explained. Genome-wide association studies have since identified eight SNPs at six loci which together account for approximately 15% of the genetic risk of TGCT and offer novel biological insights into testicular germ-cell oncogenesis. In this review, we summarize the genetic epidemiology of TGCT, detail the contribution genome-wide association studies have made to our understanding of the genetic basis of TGCT and reflect on how future technological advances may assist in revealing the remaining genetic factors underlying TGCT susceptibility. PMID- 21623832 TI - Peritubular myoid cell-derived factors and its potential role in the progression of testicular germ cell tumours. AB - The tumour surrounding stroma, known as reactive stroma, is a crucial factor to understand cancer cell growth and invasion. In the normal adult testis, the stroma contains extracellular matrix components, fibroblasts, infiltrating leucocytes, lymphocytes, macrophages and capillaries, as well as other specific cell populations, like Leydig cells and a thin myoepithelium surrounding the seminiferous tubules constituted by the peritubular cells. All these cells are an important source of proliferation and survival promoting signals, proteolytic enzymes, migratory cues and pro-angiogenic factors. Ascribable to this pro invasive activity, the tumour reactive stroma cells, especially cancer-associated myofibroblasts, have emerged as a promising target for cancer therapy. This review is focused on the potential role of the peritubular myoid cells in the development of testicular germ cell tumours as the precursors of cancer associated myofibroblast and on an experimental model for the study of testis germinal cancer stroma and on the differences between normal and tumour associated stromal cells, including the molecular mechanisms that mediate the important cancer stroma crosstalk. Special attention will be paid to the cancer associated myofibroblasts as possible therapeutic targets, because they are one of the main components of the reactive stroma and are known to secrete a variety of paracrine factors that stimulate tumour progression. PMID- 21623833 TI - Impact of classification of mixed germ-cell tumours on incidence trends of non seminoma. AB - Seminomas and non-seminomas [embryonal carcinomas, yolk sac tumours, teratomas, choriocarcinomas, mixed germ-cell tumours (MGCT)] are the major histological types of testicular germ-cell tumours (TGCT). TGCTs composed of both seminomatous and non-seminomatous elements have been coded as their non-seminoma component in the World Health Organization classification. In the late 1980s, a provisional International Classification of Diseases for Oncology (ICD-O) morphology code for MGCT was introduced. Using data from the Surveillance, Epidemiology and End Results Program and two population-based German cancer registries, we examined the impact of MGCT classification on TGCT trends. Cases were identified using ICD O topography (ICD-9: 186; ICD-10: C62) and morphology codes (seminoma=9060-9062, 9064; embryonal carcinoma=9070; yolk sack tumour=9071; teratoma=9080-9084, 9102; choriocarcinoma=9100, 9101; MGCT=9085; all non-seminoma=9065-9102). As MGCTs and teratoma are often grouped as a single histological group, we analysed teratoma both including and excluding MGCTs. Between 1988 and 2007, incidence rates of MGCT in the US increased 407%. Rates of teratoma including MGCT increased 80%, whereas rates of teratoma excluding MGCT decreased 71%. Rates of embryonal carcinoma [-40%] and choriocarcinoma [-22%] also declined, suggesting that the code for MGCT is now being used for any mixed histology. Similar declines in incidence were observed in the German comparison populations. The declines in incidence of teratoma (excluding MGCT), embryonal carcinoma and choriocarcinoma in the US data since 1988 are likely in part because of increases in classifying any TGCT with mixed histology as MGCT. These results suggest that analysis of trends in specific histological types of non-seminoma should be interpreted cautiously. PMID- 21623834 TI - Unexpected response of a captive blackeye thicklip, Hemigymnus melapterus (Bloch), from Lizard Island, Australia, exposed to juvenile isopods Gnathia aureamaculosa Ferreira & Smit. PMID- 21623835 TI - Epidemiological aspects of viral haemorrhagic septicaemia virus genotype II isolated from Baltic herring, Clupea harengus membras L. AB - This study was carried out to clarify the role of wild fish, especially Baltic herring, Clupea harengus membras L., in the epidemiology of viral haemorrhagic septicaemia virus (VHSV) in brackish water in Finland. Baltic herring with no visible signs of disease were collected from the Archipelago Sea, the Gulf of Bothnia and the eastern Gulf of Finland. In total, 7580 herring were examined by virus isolation as 758 pooled samples and 3029 wild salmonid broodfish as pooled samples during 2004-2006. VHSV was isolated from 51 pooled herring samples in bluegill fibroblast-2 cells, but not in epithelioma papulosum cyprini cells. The majority of isolations were from the coastal archipelago and from fish caught during the spawning season. Based on glycoprotein (G) gene sequences, the virus was classified as a member of genotype II of VHSV. Pairwise comparisons of the G gene regions of herring isolates revealed that all the isolates were closely related, with 98.8-100% nucleotide homology. Phylogenetic analyses revealed that they were closely related to the strains isolated previously from herring and sprat, Sprattus sprattus (L.), in Gotland and to the VHSV isolates from European river lamprey, Lampetra fluviatilis (L.), in the rivers that flow into the Bothnian Bay. The infection in Baltic herring is likely to be independent of the VHSV Id epidemic in farmed rainbow trout, Oncorhynchus mykiss (Walbaum). PMID- 21623836 TI - The effect of water temperature on vertebral deformities and vaccine-induced abdominal lesions in Atlantic salmon, Salmo salar L. AB - This study investigates the effects of water temperature (T) on vaccine-induced abdominal lesions (i.p. injection with oil-adjuvant vaccine) and vertebral deformities in Atlantic salmon. Quadruple groups of vaccinated (V) or unvaccinated (U) underyearling smolts were reared in tanks under four different temperature regimes for 6 weeks in fresh water (FW) followed by 6 weeks in sea water (SW). The four different T regimes were 10 degrees C FW-10 degrees C SW (10-10), 10 degrees C FW-16 degrees C SW (10-16), 16 degrees C FW-10 degrees C SW (16-10) and 16 degrees C FW-16 degrees C SW (16-16). After the temperature regimes were finished, the fish were group-tagged and transferred to a common sea cage for on-growth until harvest size. At termination, weight was significantly affected by both T and V, while lesion score and deformities were affected by T only. The weight difference between the largest and smallest U group was 20.3% (16-10 U: 2.4 kg, 10-16 U: 1.89 kg), while the largest difference between U and V fish within a T regime was 28.7% (16-16 U: 2.1 kg, 16-16 V: 1.5 kg). Fish from the 16-16, 16-10 and 10-16 regimes had a significant higher lesion score than those from the 10-10 regime. Fish from the 10-16 and 16-16 regimes displayed a significantly higher prevalence of vertebral deformities (palpation : 13-27%, radiology: 88-94%) than fish from the 10-10 and 16-10 regimes (palpation: 2-3%, radiology: 27-65%). Vertebra number 26 (located beneath the dorsal fin) was the most frequently affected vertebra in smolts, while vertebra number 43 (located above the anal fin) was most frequently affected in adults. PMID- 21623837 TI - Molecular and functional identification of organic anion transporter isoforms in cultured bovine mammary epithelial cells (BME-UV). AB - Mammary epithelial cells express a diversity of membrane transporters including members of organic cation and organic anion (OAT) transporter subfamilies. Four mammal OAT isoforms have been identified: OAT-1, OAT-2, OAT-3, and OAT-4. The pharmacological significance of OAT isoforms has been emphasized because of their role in the movement of a wide variety of substrates across epithelial barriers. The present study identified (molecularly and functionally) bovine OAT isoforms in bovine mammary epithelial (BME-UV) cells. mRNA expression levels of all tested transporters in BME-UV cells were less than expression levels of the corresponding transporters in bovine kidney. Directionality in the flux of P aminohippuric acid and acetylsalicylate, compounds known to interact with OAT-1 and OAT-2, respectively, across BME-UV monolayers was not observed at the concentrations used in this study. Directionality was, however, observed in the flux of estrone sulfate (EsS). Adding probenecid, penicillin G or nonradiolabeled EsS to the apical donor compartment significantly increased the apical-to basolateral flux of EsS across the BME-UV monolayer. These results suggest that BME-UV cells express an organic anion transport system, making it a potentially useful model to study the role of this transport system in the mammary epithelial barrier. PMID- 21623838 TI - Access cavity preparation training using haptic virtual reality and microcomputed tomography tooth models. AB - AIM: To evaluate the effectiveness of haptic virtual reality (VR) simulator training using microcomputed tomography (micro-CT) tooth models on minimizing procedural errors in endodontic access preparation. METHODOLOGY: Fourth year dental students underwent a pre-training assessment of access cavity preparation on an extracted maxillary molar tooth mounted on a phantom head. Students were then randomized to training on either the micro-CT tooth models with a haptic VR simulator (n = 16) or extracted teeth in a phantom head (n = 16) training environments for 3 days, after which the assessment was repeated. The main outcome measure was procedural errors assessed by an expert blinded to trainee and training status. The secondary outcome measures were tooth mass loss and task completion time. The Wilcoxon test was used to examine the differences between pre-training and post-training error scores, on the same group. The Mann-Whitney test was used to detect any differences between haptic VR training and phantom head training groups. The independent t-test was used to make a comparison on tooth mass removed and task completion time between the haptic VR training and phantom head training groups. RESULTS: Post-training performance had improved compared with pre-training performance in error scores in both groups (P < 0.05). However, error score reduction between the haptic VR simulator and the conventional training group was not significantly different (P > 0.05). The VR simulator group decreased significantly (P < 0.05) the amount of hard tissue volume lost on the post-training exercise. Task completion time was not significantly different (P > 0.05) in both groups. CONCLUSIONS: Training on the haptic VR simulator and conventional phantom head had equivalent effects on minimizing procedural errors in endodontic access cavity preparation. PMID- 21623839 TI - Unpacking the hedonic paradox: a dynamic analysis of the relationships between financial capital, social capital and life satisfaction. AB - Does money buy happiness? Or is happiness derived from looking outwards towards our social networks? Many researchers have answered these questions by exploring whether the best predictor of well-being is either economic or social (or some fixed combination of the two). This paper argues for a dynamic perspective on the capacity for economic and social factors to predict well-being. In two studies, we show that both money (individual income) and community (social capital) can be the basis for individual happiness. However, the relative influence of each factor depends on the context within which happiness is considered, and how this shapes the way people define the self. Study 1 primes either money or community in the laboratory and demonstrates that such priming shifts individual values (so that they are economic vs. communal) and determines the extent to which income is more (vs. less) predictive of life satisfaction than social relations. Study 2 looks at these same priming processes in the external world (with people travelling to vs. from work). Both studies show that while money can become the basis of happiness when the self is defined in economic terms, the role of community relations in predicting happiness is more stable across contexts. PMID- 21623840 TI - Experimental high-volume hemofiltration with predilutional tris hydroxymethylaminomethane for correction of low tidal volume ventilation-induced acidosis. AB - The most common method of controlling acidemia during lung-protective ventilation is CO2 removal with an extracorporeal lung assist (ECLA) system. Another possibility to prevent acidemia is based on intravenous (i.v.) application of tris-hydroxymethyl-aminomethane (3 mol/L, THAM) buffer, which can bind hydrogen protons and which can be removed from the body via renal replacement therapy (RRT). We investigated whether RRT combined with predilutional (prefilter) THAM application provides an alternative to ECLA for a rescue situation. For this, anesthetized pigs, 40 kg of body weight, six animals per group, underwent 5 h of acidemia (pH 7.19-7.24) induced by acid infusion and permissive hypercapnia (low tidal volume ventilation, PaCO2 80-90 mmHg). Isovolemic, high-volume hemofiltration (HVHF) was operated with predilutional THAM-infusion for treatment. To evaluate adverse effects of this approach, we set up further groups: HVHF with postdilutional (post-filter) THAM-application; i.v.-THAM without HVHF; normal pH homeostasis with HVHF. Acid-base parameters, hemodynamics, renal function, and lung morphology were investigated. HVHF with predilutional THAM-infusion of 8 mmol/kg/h allowed fast pH normalization, significant reduction in PaCO2 to 56 mmHg and tolerable hemodynamics. HVHF alone or lower dose i.v. THAM (2 mmol/kg/h) failed to produce a comparable result. A postdilutional THAM infusion reduced hemodynamic tolerability and increased lung edema formation. HVHF in pigs with normal acid-base status resulted in a decreased base excess and urine acidification. In conclusion, predilutional THAM application and HVHF corrected the acid-base disorder and improved pulmonary hemodynamics. Further studies are necessary to optimize the protocol including the dosage. PMID- 21623841 TI - Impact of oxygenator selection on hemodynamic energy indicators under pulsatile and nonpulsatile flow in a neonatal extracorporeal life support model. AB - This study compared the quality of perfusion delivered by two oxygenators--the hollow-fiber membrane Capiox Baby RX05 and silicone membrane Medtronic 0800- using hemodynamic energy indicators. The oxygenators were tested across varying flow rates and perfusion modes in a neonatal extracorporeal life support (ECLS) model. The experimental ECLS circuit included a Jostra HL-20 heart/lung machine with Jostra Roller pump, oxygenators with associated tubing and components, and a neonatal pseudo-patient. We used a 40/60 glycerin/water solution in the circuit as a blood analog. Testing occurred at flow rates of 250, 500, and 750 mL/min at 37 degrees C under both pulsatile and nonpulsatile flow conditions. Hemodynamic data points consisted of recording 20-s intervals of data, and a total of 96 experimental repetitions were conducted. The pressure drop across the Capiox Baby RX05 oxygenator was significantly lower than the pressure drop across the Medtronic 0800 at all flow rates and perfusion modes. Furthermore, the Medtronic 0800 oxygenator showed significantly lower post-oxygenator energy equivalent pressures, total hemodynamic energy values, and surplus hemodynamic energy retention values compared to those of the Capiox Baby RX05. These results indicate the Medtronic 0800 oxygenator significantly dampens the hemodynamic energy compared to the Capiox Baby RX05. Consequently, clinical use of the Medtronic 0800 in a pulsatile ECLS setting is likely to mitigate the benefits provided by pulsatile flow. In contrast, the Capiox Baby RX05 better transmits hemodynamic energy to the patient with much lower pressure drop. PMID- 21623842 TI - Clinical effectiveness of centrifugal pump to produce pulsatile flow during cardiopulmonary bypass in patients undergoing cardiac surgery. PMID- 21623844 TI - Enlightening the life sciences: the history of halobacterial and microbial rhodopsin research. AB - The history of research on microbial rhodopsins offers a novel perspective on the history of the molecular life sciences. Events in this history play important roles in the development of fields such as general microbiology, membrane research, bioenergetics, metagenomics and, very recently, neurobiology. New concepts, techniques, methods and fields have arisen as a result of microbial rhodopsin investigations. In addition, the history of microbial rhodopsins sheds light on the dynamic connections between basic and applied science, and hypothesis-driven and data-driven approaches. The story begins with the late nineteenth century discovery of microorganisms on salted fish and leads into ecological and taxonomical studies of halobacteria in hypersaline environments. These programmes were built on by the discovery of bacteriorhodopsin in organisms that are part of what is now known as the archaeal genus Halobacterium. The transfer of techniques from bacteriorhodopsin studies to the metagenomic discovery of proteorhodopsin in 2000 further extended the field. Microbial rhodopsins have also been used as model systems to understand membrane protein structure and function, and they have become the target of technological applications such as optogenetics and nanotechnology. Analysing the connections between these historical episodes provides a rich example of how science works over longer time periods, especially with regard to the transfer of materials, methods and concepts between different research fields. PMID- 21623845 TI - Agricultural land-use affects the nutritional quality of stream microbial communities. AB - We investigated how the lipid composition (fatty acids and sterols) of benthic microbial mats, which represent an important basal food resource for stream food webs, differs between tropical streams located in protected pristine and agricultural Cerrado savannah areas. The total microbial biomass and lipid composition differed significantly between pristine and agricultural streams in parallel with differences in water quality and hydrodynamic characteristics. Agricultural streams exhibited lower total biomass of benthic microbial mats than pristine streams. However, the higher concentrations of essential polyunsaturated fatty acids, such as linoleic acid (LIN, 18:2omega6), alpha-linolenic acid (ALA, 18:3omega3), and eicosapentaenoic acid (EPA, 20:5omega3), that were observed in agricultural streams suggest enhanced lipid complexity and a higher nutritional quality of the microbial community relative to pristine streams. Meanwhile, pristine stream microbial communities had higher total concentrations of saturated fatty acids and cholesterol than those of agricultural streams, reflecting their heterotrophic microbial communities. Moreover, stream morphotype and associated differences in the hydrodynamic characteristics affected the community composition and thereby also the lipid composition of microbial mats. Land-use-induced changes in the total biomass and lipid composition of microbial communities may affect the trophic transfer of energy in stream food webs, leading to changes in the composition and productivity of primary consumers and their predators, and thereby affecting stream ecosystem functioning. PMID- 21623846 TI - Enhancement of immunoregulatory effects of Lactobacillus gasseri TMC0356 by heat treatment and culture medium. AB - AIMS: The aim of this study was to investigate the influence of heat treatment and culture media on the immunoregulatory effects of a probiotic strain, Lactobacillus gasseri TMC0356 (TMC0356). METHODS AND RESULTS: TMC0356 cultured in deMan-Rogosa-Sharpe and same food grade (FG) media were inactivated with the heat treatment at 70 and 90 degrees C. Viable and heat-killed TMC0356 were tested for their ability to induce interleukin (IL)-12 production in the murine macrophage cell line J774.1. These TMC0356 were examined for their resistance to N acetylmuramidase. Their morphology was observed by scanning electron microscopy. The heat-killed TMC0356 significantly induced IL-12 production in J774.1 cells and exhibited enhanced resistance to N-acetylmuramidase compared with viable TMC0356. Morphological changes were observed in TMC0356 when cultured in FG medium. Cell morphology and induction of IL-12 production in J774.1 cells were also associated. CONCLUSIONS: These results suggest that heat treatment and culture medium composition modified the immunoregulatory effects of TMC0356 to induce IL-12 production in macrophages. SIGNIFICANCE AND IMPACT OF THE STUDY: These results demonstrate that probiotic immunoregulatory effects may be modified by the processing technology of cell preparation. PMID- 21623847 TI - Differential growth of Legionella pneumophila strains within a range of amoebae at various temperatures associated with in-premise plumbing. AB - AIMS: The potential effect of in-premise plumbing temperatures (24, 32, 37 and 41 degrees C) on the growth of five different Legionella pneumophila strains within free-living amoebae (Acanthamoeba polyphaga, Hartmannella vermiformis and Naegleria fowleri) was examined. METHODS AND RESULTS: Compared with controls that actively fed on Escherichia coli prey, when Leg. pneumophila was used as prey, strains Lp02 and Bloomington-2 increased in growth at 30, 32 and 37 degrees C while strains Philadelphia-1 and Chicago 2 did not grow at any temperature within A. polyphaga. Strains Lp02, Bloomington-2 and Dallas 1E did not proliferate in the presence of H. vermiformis nor did strain Philadelphia-1 in the presence of N. fowleri. Yet, strain Bloomington-2 grew at all temperatures examined within N. fowleri, while strain Lp02 proliferated at all temperatures except 41 degrees C. More intriguing, strain Chicago 2 only grew at 32 degrees C within H. vermiformis and N. fowleri suggesting a limited temperature growth range for this strain. CONCLUSIONS: Identifying the presence of pathogenic legionellae may require the use of multiple host amoebae and incubation temperatures. SIGNIFICANCE AND IMPACT OF THE STUDY: Temperature conditions and species of amoeba host supported in drinking water appear to be important for the selection of human-pathogenic legionellae and point to future research required to better understand Legionella ecology. PMID- 21623848 TI - Chlorine dioxide inactivation of bacterial threat agents. AB - AIMS: To evaluate the efficacy of chlorine dioxide (ClO(2)) against seven species of bacterial threat (BT) agents in water. METHODS AND RESULTS: Two strains of Bacillus anthracis spores, Yersinia pestis, Francisella tularensis, Burkholderia pseudomallei, Burkholderia mallei and Brucella species were each inoculated into a ClO(2) solution with an initial concentration of 2.0 (spores only) and 0.25 mg l(-1) (all other bacteria) at pH 7 or 8, 5 or 25 degrees C. At 0.25 mg l(-1) in potable water, six species were inactivated by at least three orders of magnitude within 10 min. Bacillus anthracis spores required up to 7 h at 5 degrees C for the same inactivation with 2.0 mg l(-1) ClO(2). CONCLUSIONS: Typical ClO(2) doses used in water treatment facilities would be effective against all bacteria tested except B. anthracis spores that would require up to 7 h with the largest allowable dose of 2 mg l(-1) ClO(2). Other water treatment processes may be required in addition to ClO(2) disinfection for effective spore removal or inactivation. SIGNIFICANCE AND IMPACT OF STUDY: The data obtained from this study provide valuable information for water treatment facilities and public health officials in the event that a potable water supply is contaminated with these BT agents. PMID- 21623849 TI - Use of fruit residues for pectinase production by Aspergillus flavipes FP-500 and Aspergillus terreus FP-370. AB - AIMS: Utilization of fruit residues for pectinase production by two Aspergillus strains for recognizing the effects of some factors during fermentation and describing enzyme production kinetics. METHODS AND RESULTS: Pectinase production on several fruit residues was compared. The effects of three factors on the production of several pectinases were evaluated by a full factorial 2(k) experimental design. Higher activities were obtained on lemon peel. In both strains, acidic pH values and high carbon source concentration favoured exopectinase and endopectinase production, while higher pH values and low carbon source concentration promoted pectin lyase and rhamnogalacturonase production. Unstructured mathematical modelling provided a good description of pectinase production in a submerged batch culture. CONCLUSIONS: Fruit residues were very good substrates for pectinase production, and Aspergillus strains used showed a promising performance in submerged fermentation. Mathematical modelling was useful to describe growth and pectinase production. SIGNIFICANCE AND IMPACT OF THE STUDY: Lemon peel can be used as a substrate to obtain high pectinase titres by Aspergillus flavipes FP-500 and Aspergillus terreus FP-370. The factors that contributed to improve the yield were identified, which supports the possibility of using this substrate in the industrial production of these enzymes. PMID- 21623850 TI - Systematic review: epidemiology of hepatitis C genotype 6 and its management. AB - BACKGROUND: Hepatitis C virus (HCV) genotype 6 is common among patients from Southeast Asia and the surrounding regions, where HCV prevalence is also high. HCV genotype 6 has great genetic diversity and different response to antiviral therapy compared with the more commonly known genotype 1. AIM: Our goal was to provide a systematic review of the current literature on the epidemiology, classification and treatment of HCV genotype 6. METHODS: A search using PubMed for 'hepatitis C' AND 'genotype 6' produced a total of 47 articles, of which 33 articles were found to be relevant and included in this review. Additional articles were identified using the reference lists of these 33 primary articles. RESULTS: The prevalence of HCV genotype 6 is estimated to be as high as 50% in some regions of Southeast Asia with demonstrated significance among intravenous drug users and thalassemia major patients. Although previous line probe assays may have misclassified HCV genotype 6 as genotype 1, newer line probe assays can more accurately and reliably determine HCV genotype. Patients infected with HCV genotype 6 respond better to interferon-based therapy compared with those infected with genotype 1, although patient baseline clinical characteristics and side effect profiles are similar between HCV genotype 6 and other HCV genotypes. CONCLUSIONS: Future studies should seek to clarify issues regarding early predictors for treatment response in patients with HCV genotype 6, and the impact of ethnic and genotypic factors to treatment response in HCV genotype 6 patients. PMID- 21623851 TI - Meta-analysis: insulin resistance and sustained virological response in hepatitis C. AB - BACKGROUND: A higher baseline homeostasis model assessment of insulin resistance (HOMA-IR) score has sometimes predicted a poorer sustained virological response (SVR) rate to peginterferon/ribavirin therapy in treatment-naive chronic hepatitis C patients. AIM: To perform a meta-analysis to evaluate the impact of HOMA-IR on SVR in hepatitis C. METHODS: Relevant studies were identified by searching Medline and EMBASE. We identified 17 publications that addressed the influence of insulin resistance on SVR. The random effect model of Der Simonian and Laird method were used for heterogeneous studies using the Meta-Disc software 1.4, Madrid, Spain. RESULTS: Normal insulin sensitivity was associated with a higher rate of SVR [odds ratio (OR) 2.86 (95%CI: 1.97-4.16)] in comparison with insulin resistance. Moreover, in separate analysis by genotype selecting studies that used HOMA-IR > 2 as cut-off defining insulin resistance, SVR was higher in patients with HOMA-IR < 2 in all genotypes: HCV-1 [OR: 2.16 (95%CI: 1.51-3.08)], HCV-2&3 [OR: 3.06 (95%CI: 1.06-8.82)] and HCV-4 [OR: 6.65(95%CI: 2.51-17.61)]. Studies reporting no association between HOMA and SVR included easy-to-cure cohorts, analysed variables strongly related with insulin resistance like body mass index, steatosis, hyper gammaGT, age and fibrosis and reported differences in handling and interpretation of HOMA-IR. CONCLUSION: Elevated HOMA-IR was associated with a lower cure rate of patients with hepatitis C treated with Peg IFN-alpha/ribavirin irrespective of genotype, and the more difficult-to-treat cohort, the better the HOMA-IR prediction. HOMA-IR is, as a surrogate marker of insulin resistance, susceptible to some biases derived from both handling and interpretation. PMID- 21623852 TI - Systematic review: the epidemiology and natural history of non-alcoholic fatty liver disease and non-alcoholic steatohepatitis in adults. AB - BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is a common cause of chronic liver disease, and its worldwide prevalence continues to increase with the growing obesity epidemic. This study assesses the epidemiology of NAFLD in adults based on clinical literature published over the past 30 years. AIM: To review epidemiology and natural history of non-alcoholic fatty liver disease and non-alcoholic steatohepatitis in adults based on clinical literature published over the past 30 years. METHODS: An in-depth search of PubMed (1980-2010) was based on five search terms: 'non-alcoholic fatty liver disease' OR 'non-alcoholic steatohepatitis' OR 'fatty liver' OR 'steatosis' AND 'incidence' [MeSH Terms] OR 'prevalence' [MeSH Terms] OR 'natural history'. Studies of paediatric cohorts were excluded. Articles were categorised by topic and summarised, noting generalisations concerning their content. RESULTS: Four study categories included NAFLD incidence, prevalence, risk factors and natural history. Studies related to NAFLD prevalence and incidence indicate that the diagnosis is heterogeneous and relies on a variety of assessment tools, including liver biopsy, radiological tests such as ultrasonography, and blood testing such as liver enzymes. The prevalence of NAFLD is highest in populations with pre-existing metabolic conditions such as obesity and type II diabetes. Many studies investigating the natural history of NAFLD verify the progression from NASH to advanced fibrosis and hepatocellular carcinoma. CONCLUSIONS: Non-alcoholic fatty liver disease is the most common cause of elevated liver enzymes. Within the NAFLD spectrum, only NASH progresses to cirrhosis and hepatocellular carcinoma. With the growing epidemic of obesity, the prevalence and impact of NAFLD continues to increase, making NASH potentially the most common cause of advanced liver disease in coming decades. PMID- 21623853 TI - Fear of falling from a daily life perspective; narratives from later life. AB - Fear of falling is a well-known condition in later life. The aim of this study was to illuminate the experiences and the meaning of fear of falling in a daily life context. The method used was a qualitative study inspired by interpretive phenomenology. In narrative interviews, five community-dwelling women over 80 years of age told about their fear of falling from a daily-life perspective. The overall thematic analysis resulted in three main themes: the meaning of managing daily life necessities; keeping in contact with the outside; living with fear. The findings showed that to live with fear of falling was to discipline daily life, and to learn to live with the challenge of a vulnerable bodily condition and of losing control at different levels: from falling, from incontinence, from dirt and from the stigma of being in a humiliating situation. The women created a perception of independence while they were dependent on help and community care and on news from the outside. At an existential level, they coped with their fear by strengthening their will. The conclusion was that the older women studied accepted the condition of fear of falling. They shared the ability to cope in various ways with the limitations of their bodily capacity and their imbalance. PMID- 21623854 TI - Different relationships between the first 2 years on growth hormone treatment and the d3-growth hormone receptor polymorphism in short small-for-gestational-age (SGA) children. AB - BACKGROUND: There has been controversy in recent years on whether the d3 polymorphism of the GH receptor is associated with a better growth response to GH in idiopathic short children born small for gestational age (SGA). METHODS: In this prospective study, we evaluated exon 3-GHR polymorphisms in 142 (62 f, 80 m) short prepubertal children born SGA (birth length and/or weight of <=-2 SD for GA) and treated with rhGH (mean dose of 0.30 mg/kg/week) in 24 centres in Germany. A growth prediction for the first year of therapy was calculated for each child according to Ranke and co-workers. The index of responsiveness (IOR) was calculated by dividing the response (observed growth minus predicted growth) by the standard error of the prediction. All analyses were performed in one centre on samples collected and shipped on filter paper. The DNA fragment containing or missing exon 3 of the GHR was amplified by multiplex PCR. RESULTS: The fl-GHR isoform was most common with a frequency of 47.8%, followed by the d3/fl isoform with 38% and the d3-GHR isoform with 14.2%. There were no significant differences regarding gestational age, birth weight and birth length, mid parental height-SDS, chronological age at start of therapy, height-SDS, BMI SDS, height velocity and GH dose between the different subgroups according to the genotype. After the first treatment year, height (H)-SDS (P < 0.05), height velocity (HV) (P < 0.01), HV-SDS (P < 0.001) and delta-H-SDS (P < 0.05) were significantly higher in patients with d3-GHR than in those with fl-GHR. The mean IOR was above 0 in children with at least one d3 allele, and highest, with 0.54, in those with the d3-GHR isoform. After the second year on GH, no differences between the different GHR-isoforms were found. CONCLUSIONS: According to our results, the exon 3-deleted GHR explains the better growth response to GH only for the first and not for the second year. PMID- 21623855 TI - Hyperthyroidism and female urinary incontinence: a population-based cohort study. AB - OBJECTIVE: The imbalanced autonomic nervous system present in hyperthyroidism may cause lower urinary tract symptoms. Urinary incontinence (UI) is the most bothersome lower urinary tract symptom; however, in the literature, reports regarding urinary dysfunction and/or incontinence among hyperthyroid patients are scarce. This population-based cohort study aimed to examine the relationship between hyperthyroidism in women and the risk of developing UI in Taiwan. DESIGN: This study used data from the Longitudinal Health Insurance Database. For this study, 10,817 female patients diagnosed with hyperthyroidism from 2001 to 2005 were recruited together with a comparison cohort of 54,085 matched enrollees who did not have a history of hyperthyroidism. All patients were tracked for a 3-year period from their index date to identify those who had a subsequent UI. The stratified Cox proportional hazards models were used to compute the risk of UI between study and comparison cohorts. RESULTS: During the follow-up period, of 64,169 patients, 173 (1.60%) from the hyperthyroidism group and 560 (1.04%) from the comparison group, had a diagnosis of UI. The regression analysis showed that, after adjusting for monthly income, geographic region, urbanization level of the community in which the patient resided, obesity and hysterectomy, patients with hyperthyroidism were more likely to have UI during the 3-year follow-up period than the comparison patients (hazard ratio = 1.54; 95% CI = 1.30-1.83; P < 0.001). CONCLUSION: Our results suggest an increased risk of UI in patients with hyperthyroidism at the 3-year follow-up. PMID- 21623856 TI - High serum isoflavone concentrations are associated with the risk of precocious puberty in Korean girls. AB - OBJECTIVE: The gradual decrease in the age of onset of puberty raises concerns about the contribution of phytoestrogen intake on sexual maturation. However, no data are available on the association between serum isoflavone (genistein, daidzein) concentration and central precocious puberty (CPP). The aim of the study was to test the association between serum isoflavone concentrations and the risk of CPP in Korean girls. STUDY DESIGN: A case-control study was conducted at Inje University Hospital in Korea. One-hundred and eight girls with CPP (aged 8.6 +/- 0.8 year) and 91 age-matched controls (aged 8.5 +/- 0.8 year) were examined. RESULTS: Serum concentrations of daidzein (P = 0.0202), genistein (P = 0.0021) and total isoflavones (P = 0.0009) were higher in children with CPP than in normal children. When the children were categorized into three groups according to total serum isoflavone as follows: <30, 30-70 and >=70 nmol/l, serum concentrations of daidzein, genistein and total isoflavones significantly increased across three categories of serum isoflavone (P < 0.0001). The prevalence of CPP was significantly higher in children with serum isoflavone level of >=30 nmol/l than those with serum level of <30 nmol/l (P = 0.0008). The adjusted OR for precocious puberty increased significantly across a range of total serum isoflavone (OR = 4.39; 95% CI: 1.83-10.51 vs OR = 5.22; 95% CI: 2.07 13.20; P = 0.001). CONCLUSION: These results suggest that elevated serum isoflavones may be associated with the risk of CPP in Korean girls. PMID- 21623857 TI - The increased incidence of congenital hypothyroidism: fact or fancy? AB - OBJECTIVE: The incidence of congenital hypothyroidism (CH) detected by newborn screening in the US has increased significantly since the early 1990s. We defined the characteristics associated with the increased incidence. PATIENTS: A cohort of children with CH born during an earlier period of low incidence (1991-94) was compared with a cohort born during a later period when the incidence of CH had doubled (2001-04). MEASUREMENTS: Screening was performed with T4 as the primary marker and thyroid stimulating hormone (TSH) on selected specimens. Follow-up on hypothyroid children determined whether they had permanent or transient hypothyroidism. Cases were classified based on laboratory results: initial TSH >=100 mU/l was 'severe,' initial TSH <100 mU/l but >=20 mU/l was 'mild' and initial TSH <20 mU/l with subsequent abnormal TSH was 'delayed'. RESULTS: The overall incidence of CH almost doubled between the two time periods, from 1:3010 to 1:1660. Excess cases were found in the mild and delayed categories, with no increase in severe cases. The proportion of transient cases was <5% in severe cases, 40% in mild cases and 70% among delayed cases. There was no difference in the proportion of transient case between the two time periods. Modifications to the T4/TSH testing protocol between the two time periods resulted in substantially increased numbers of specimens in the younger cohort being selected for TSH testing in both initial and repeat specimens. CONCLUSION: The rising incidence of CH in Massachusetts is confined to mild and delayed cases. Our findings suggest that this rise is attributable to enhanced detection rather than an absolute increase in numbers. PMID- 21623858 TI - Rapid decrease in adrenal responsiveness to ACTH stimulation after successful pituitary surgery in patients with Cushing's disease. AB - OBJECTIVE: The aim of this study was to investigate the effects of transsphenoidal surgery (TS) on the adrenal sensitivity to ACTH (adrenocorticotropin) stimulation in patients with Cushing's disease (CD). METHODS: We measured the cortisol response to 1 MUg synthetic ACTH (1-24) 6 days after pituitary surgery in 45 patients with CD. Mean follow-up period was 56.5 months (SE 4.7). RESULTS: In 24 of 28 patients in sustained remission after pituitary surgery, peak cortisol concentrations below 774 nm (28.0 MUg/dl) were recorded after stimulation with 1 MUg synthetic ACTH (86%). Two patients with recurrent disease after initial remission (late relapse) also showed ACTH stimulated peak cortisol levels below 774 nM. Fourteen of 15 patients with persistent CD after surgery (early failure) showed absolute peak cortisol levels >774 nm in response to ACTH stimulation. CONCLUSION: Patients in remission after pituitary surgery for CD showed a rapid decrease of adrenal responsiveness to exogenous ACTH stimulation. This phenomenon may be explained by ACTH-receptor down-regulation in the adrenal cortex after complete removal of the pituitary corticotroph adenoma. In our study, the postoperative low-dose ACTH stimulation test had a sensitivity of 93% and a specificity of 87% in predicting immediate remission of CD after pituitary surgery. PMID- 21623859 TI - Aortic root ectasia in patients with acromegaly: an emerging complication. PMID- 21623860 TI - Subcutaneous adipose tissue thickness in adults - correlation with BMI and recommendations for pen needle lengths for subcutaneous self-injection. AB - OBJECTIVE: One of the aims of a subcutaneous (SC) injection is to avoid intradermal or intramuscular injections. Pen systems are an alternative solution to single-use syringes and have become standard for example diabetes therapy. Shorter and smaller needles minimize pain and the risk of intramuscular injections. The thickness of subcutaneous adipose tissue (SCAT) varies with position and with body mass index (BMI). The aim of this study was the creation of a map of SCAT thickness at typical spots for SC self-injection. MATERIALS AND METHODS: MRI scans of 116 prospectively enroled volunteers (56 men and 60 women) were analysed. SCAT thickness was determined at 17 spots over the abdominal wall, left thigh, buttocks and upper arm, typical sites for subcutaneous self injection. SCAT thicknesses were correlated with BMI and waist-to-hip ratio (WHR), and a linear curve fit was performed. The best fitting linear functions for the prediction of the SCAT thickness dependent on BMI and WHR were derived. RESULTS: Correlations between SCAT and BMI were higher (0.67-0.21) than with WHR (-0.67 to 0.09). In women, correlation coefficients between SCAT data at the abdomen and BMI/WHR were higher than in men. On the other hand, data showed better correlations at the extremities in men. CONCLUSIONS: The data, with correlation between BMI and fat thickness at different injection sites in relation to gender, provide guidance in selecting an adequate pen needle length for deep and safe subcutaneous self-injection. WHR was a much weaker predictor when compared to BMI. PMID- 21623861 TI - Regulation of glucose metabolism and the skeleton. AB - Complex interactions occur among adipose tissue, the central nervous system, bone and pancreas to integrate bone remodelling, glucose, lipid and energy metabolism. Data obtained largely from the judicious use of gain-of-function and loss-of function genetic mouse models show that leptin, an adipocyte-secreted product, indirectly inhibits bone accrual through a central pathway comprising the hypothalamus and central nervous system. Increased sympathetic output acting via beta2-adrenergic receptors present in osteoblasts decreases bone formation and causes increased bone resorption. Insulin is a key molecular link between bone remodelling and energy metabolism. Insulin signalling in the osteoblasts increases bone formation and resorption as well as the release of undercarboxylated osteocalcin. An increase in the release of bone-derived undercarboxylated osteocalcin into the systemic circulation enables it to act as a circulating hormone to stimulate insulin production and secretion by pancreatic beta-cells and adiponectin by adipocytes. Insulin sensitivity increases, lipolysis and fat accumulation decreases while energy expenditure increases. Whether this model of integrative physiology involving the skeleton, pancreas and adipose tissue, so elegantly demonstrated in rodents, is applicable to humans is controversial. The mouse Esp gene, encoding an intracellular tyrosine phosphatase that negatively regulates insulin signalling in osteoblasts, is a pseudogene in humans, and a homolog for the Esp gene has so far not been identified in humans. A close homologue of Esp, PTP1B, is expressed in human osteoblasts and could take the role of Esp in humans. Data available from the limited number of clinical studies do not provide a sufficient body of evidence to determine whether osteocalcin or undercarboxylated osteocalcin affects glucose metabolism in humans. PMID- 21623862 TI - A mechanism for the action of the compound DA-6034 on NF-kappaB pathway activation in Helicobacter pylori-infected gastric epithelial cells. AB - DA-6034 is a synthetic derivative of eupatilin, a flavonoid with anti inflammatory effects. The aim of this study was to investigate the effects of DA 6034 on the interactions between IkappaB kinase (IKK) and heat shock protein 90 (Hsp90), and activation of the nuclear factor-kappaB (NF-kappaB) signalling pathway in human gastric epithelial cells infected with Helicobacter pylori. MKN 45 gastric epithelial cell line was treated with DA-6034 and H. pylori. DA-6034 significantly inhibited NF-kappaB activation and upregulated the expressions of interleukin-8 (IL-8) and monocyte chemoattractant protein-1 in MKN-45 cells infected with H. pylori. However, DA-6034 did not influence activator protein-1 DNA binding activity in H. pylori-infected gastric epithelial cells. Pretreatment with DA-6034 attenuated the H. pylori-induced increase in IKK activity, and Hsp90 was associated with IKK-alpha and IKK-gamma in MKN-45 cells. Treatment with DA 6034 dissociated the Hsp90 and IKK-gamma complex in H. pylori-infected cells, leading to the inhibition of IL-8 expression. These results suggest that the eupatilin derivative 7-carboxymethyloxy-3',4',5-trimethoxy flavone has anti inflammatory activity in gastric epithelial cells infected with H. pylori through the promotion of the dissociation of the IKK-gamma-Hsp90 complex and suppression of NF-kappaB signalling. PMID- 21623863 TI - The risk of temporomandibular disorder in patients with depression: a population based cohort study. AB - OBJECTIVES: This study used a population-based retrospective cohort design to examine whether depression is a risk factor of temporomandibular disorder (TMD). METHODS: From a universal insurance database, we identified 7587 patients who are newly diagnosed individuals with depression in 2000 and 2001. A total of 30,197 comparison subjects were randomly selected from a nondepression cohort. Both groups were followed until the end of 2008 to measure the incidence of TMD. RESULTS: The incidence of TMD was 2.65 times higher in the depression cohort than in the nondepression cohort (6.16 versus 2.32 per 1000 person-years). The hazard ratio (HR) measured by multivariate Cox's proportional hazard regression analysis of TMD for the depression cohort was 2.21 (95% confidence interval (CI) 1.83 2.66), after controlling for socio-demographic factors and other psychiatric comorbidities. Women had higher risk to develop TMD than men (HR 1.61, 95% CI 1.36-1.92 for women without depression; HR 3.54, 95% CI 2.81-4.45 for women with depression). CONCLUSIONS: This study demonstrates that patients with depression are at an elevated risk of developing TMD. PMID- 21623864 TI - Equity, social determinants and public health programmes--the case of oral health. AB - The WHO Commission on Social Determinants of Health issued the 2008 report 'Closing the gap within a generation - health equity through action on the social determinants of health' in response to the widening gaps, within and between countries, in income levels, opportunities, life expectancy, health status, and access to health care. Most individuals and societies, irrespective of their philosophical and ideological stance, have limits as to how much unfairness is acceptable. In 2010, WHO published another important report on 'Equity, Social Determinants and Public Health Programmes', with the aim of translating knowledge into concrete, workable actions. Poor oral health was flagged as a severe public health problem. Oral disease and illness remain global problems and widening inequities in oral health status exist among different social groupings between and within countries. The good news is that means are available for breaking poverty and reduce if not eliminate social inequalities in oral health. Whether public health actions are initiated simply depends on the political will. The Ottawa Charter for Health Promotion (1986) and subsequent charters have emphasized the importance of policy for health, healthy environments, healthy lifestyles, and the need for orientation of health services towards health promotion and disease prevention. This report advocates that oral health for all can be promoted effectively by applying this philosophy and some major public health actions are outlined. PMID- 21623865 TI - Myofibroblastic differentiation in atypical fibroxanthomas occurring on sun exposed skin and in a burn scar: an ultrastructural and immunohistochemical study. AB - Herein, we report the investigation of two cases of atypical fibroxanthoma (AFX). One AFX developed within actinically damaged skin, as is typical, while the other developed within a burn scar within non-sun-exposed skin. The two tumors showed almost identical histopathological, immunohistochemical and ultrastructural features. The tumors were composed of pleomorphic spindled, epithelioid, multinucleated and bizarre cells with enlarged atypical nuclei. Most tumor cells expressed vimentin and about 50% expressed CD10. Some tumor cells also expressed alpha-smooth muscle actin and CD68. However, there was no expression of cytokeratins, p63, S-100 protein, melan-A, HMB 45, desmin, epithelial membrane antigen or CD34. Ultrastructurally, the tumor cells contained myofilaments with dense patches but lacked plasmalemmal caveolae and basal lamina. The most prominent finding was the identification of fibronexus junctions. In addition, there were tumor cells containing numerous lysosomal granules. In conclusion, we clearly showed myofibroblastic differentiation in AFX by electron microscopy. We report also a case of AFX directly developing within a burn scar in the absence of actinic damage. PMID- 21623866 TI - Primary cutaneous CD20-positive T-cell lymphoma. AB - CD20 is a transmembrane protein that is expressed by B cells during their development and is, therefore, commonly used to label cells of B lineage in lymphoid infiltrates. CD20-positive T-cell lymphoma is infrequent but well recognized. Cases reported in the literature show a variety of clinical and histoimmunochemical profiles. Primary cutaneous CD20-positive T-cell lymphoma is vanishingly rare; only eight cases have been previously reported. We present two new cases of this entity and describe their clinical, histological and immunohistochemical features. CD20 is a highly specific B-cell marker. However, it has been reported in a subset of normal T-cells in peripheral blood and bone marrow of healthy individuals. This subset of T-cells also expresses more often CD8 and g/d than the CD20-negative T-cells. Two main theories have been postulated to explain the expression of CD20 by neoplastic T-cells. The first possibility is that these lymphomas develop from the CD20-positive subset of normal T-cells. The second theory regards CD20 as an activation marker. Prognostic implications and therapeutic options of T-cell lymphomas with positivity for CD20 remain to be elucidated. PMID- 21623867 TI - Diagnostic utility of low-affinity nerve growth factor receptor (P 75) immunostaining in atypical fibroxanthoma. AB - BACKGROUND: Atypical fibroxanthoma (AFX) is a locally destructive, dermal-based, fibrohistiocytic, mesenchymal tumor. Immunohistochemistry helps to differentiate AFX from squamous cell carcinoma and spindle cell melanoma. Immunomarkers against p75 yield positive stains in spindled cell melanomas and negative stains in AFX, suggesting that these may be useful in differentiating these two entities. However, a recent study concluded that p75 is not a specific marker of neuroectodermal tumors; furthermore, p75 staining in AFX has only been evaluated in a few cases. METHODS: We stained 20 AFXs for p75 and various other markers. RESULTS: Reactivity was noted for vimentin (20 of 20 cases), CD10 (17/20), CD68 (14/20), CD99 (13/20), D2-40 (10/20) and p75 (1/20). CONCLUSIONS: We confirmed that CD99 and CD10 are frequently expressed in AFX (65 and 85%, respectively) and that CD31 rarely stains positive (5%). The 50% positivity rate of D2-40, in contrast with published evidence for its absence in melanoma, suggests that D2-40 may be useful for distinguishing AFX from melanoma. Furthermore, because only one sample was positive for p75, we confirm that p75 is useful in differentiating AFX from spindle cell melanoma. We advocate adding p75 and D2-40 to assist in differentiating AFX from melanoma. PMID- 21623868 TI - Angioinvasive Fusarium and concomitant Enterococcus infection arising in association with leukemia cutis. AB - Leukemia cutis represents the cutaneous infiltration of neoplastic leukocytes or their precursors that results in clinically identifiable skin lesions. For patients with myelodysplastic syndrome, developing such a lesion may indicate impending leukemic transition. These patients are also often immunocompromised, putting them at risk for infection by opportunistic fungal pathogens such as Fusarium. We describe an 85-year-old man with myelodysplastic syndrome who presented with a reddish-purple nodule with a surrounding erythematous plaque on his shin. Histopathologic examination revealed a dense diffuse infiltrate of large atypical cells in the reticular dermis, with ulceration and necrosis. Immunohistochemical studies showed positive staining with CD15, CD68 and myeloperoxidase of constituent large cells. Concurrently, there were branching and septate hyphae with occasional macroconidia-like structures throughout the infiltrate. Cultures from this lesion grew Fusarium and Enterococcus, supporting the diagnosis: leukemia cutis with superinfection involving both Fusarium and Enterococcus. To our knowledge, this is a novel report of two separate infections occurring in a lesion of leukemia cutis. This case shows that in patients with a hematologic malignancy and skin lesions, a high index of suspicion for infection is necessary when reviewing both the clinical and histopathological data to avoid overlooking an important occult infectious agent. PMID- 21623869 TI - Epstein-Barr virus-associated leiomyosarcoma with cutaneous involvement in an African child with human immunodeficiency virus: a case report and review of the literature. AB - Epstein-Barr virus (EBV)-associated smooth muscle tumors (SMTs) are infrequently encountered lesions restricted to immunocompromised patients. However, they represent the second most common tumor in children with human immunodeficiency virus (HIV) infection. We report a case of a progressively enlarging abdominal mass with cutaneous involvement in an HIV-infected, 4-year-old African girl in Malawi with clinical acquired immunodeficiency syndrome on highly active antiretroviral therapy. Analysis of an excisional specimen revealed a well differentiated leiomyosarcoma and subsequent studies revealed diffuse nuclear positivity for Epstein-Barr virus early RNAs in lesional cells. We present a report of this case and provide a summary of the literature regarding SMTs in pediatric HIV-infected patients. In addition, we draw attention to the cutaneous manifestations of SMTs in immunosuppressed populations. We highlight EBV-related SMTs as a potential cutaneous complication of HIV infection in children and immunosuppressed patients populations as a whole. PMID- 21623870 TI - Primary cutaneous marginal zone lymphoma with subclinical cutaneous involvement and biclonality. AB - Primary cutaneous extranodal marginal zone lymphoma (MZL) of mucosa-associated lymphoid tissue (MALT) represents a monoclonal B-cell neoplasm that typically presents with papules, plaques or nodules. We describe a patient with a primary cutaneous MALT lymphoma with unusual clinical features and an unusual immunophenotype. Conventional microscopy together with immunohistochemistry and in-situ hybridization showed the presence of lymphoma in normal-appearing and minimally erythematous skin as well as in clinically involved skin. Furthermore, at least two distinct clones were shown, one of which had kappa-light chain restriction, and the other of which had lambda-light chain restriction. This case represents a newly described clinical appearance of primary cutaneous MZL and shows that some patients may have more than one neoplastic clone. PMID- 21623871 TI - The mediating role of hyperactivity and inattention on sex differences in paediatric injury risk. AB - BACKGROUND: Extensive research suggests that risk of injury is higher among young boys versus young girls. The present study examined a mediational model to identify mechanisms that may explain differences in injury risk. METHODS: Reports of child behaviour and two indices of injury risk among 114 children in early childhood were obtained from parents in community-based paediatric medical centres. RESULTS: Regression analyses and post-hoc examination of indirect effects supported a mediation model in which the relationship between child sex and child injury risk was explained by hyperactivity and inattention. CONCLUSIONS: Interventions that promote child well-being by targeting constellations of externalizing behaviour problems may simultaneously decrease paediatric injury risk. PMID- 21623872 TI - Parents' experiences of caring for a child with a cleft lip and/or palate: a review of the literature. AB - This review brings together for the first time the existing quantitative and qualitative research evidence about the experiences of parents caring for a child with a cleft. It summarizes salient themes on the emotional, social and service related experiences of parents and critiques the literature to date, comparing it with wider, selected literature from the field of children's long-term conditions, including disability. The review suggests that there are similarities and differences between the literatures, in terms of research focus and approach. Similarities are found across children's conditions in the perspectives of parents on emotional, social and service-related aspects, although much of the cleft literature is focused on the early stages of children's lives. However, the quality of cleft research to date about parents' experiences has also been variable, with a narrow emphasis on cross-sectional, deficit-orientated psychological approaches focused mainly on mothers. Despite a substantial literature, little qualitative research has examined parents' perspectives in depth, particularly about their child's treatment journey. This contrasts with the wider children's literature, which has traditionally drawn not only on psychological approaches but also on the broader perspectives of sociology, social policy, nursing and health services research, using both qualitative and quantitative methods, often in integrated ways. Such approaches have been able to highlight a greater range of experiences from both mothers and fathers, about caring for a child with a long-term condition and views about treatment. The review identifies a lack of comparable research in the cleft field to examine parents' experiences and needs at different stages of their children's lives. Above all, research is needed to investigate how both mothers and fathers might experience the long-term and complex treatment journey as children become older and to elicit their views about decision making for cleft treatments, particularly elective surgeries. PMID- 21623874 TI - Tolvaptan. PMID- 21623875 TI - Successful cutaneous delivery of the photosensitizer silicon phthalocyanine 4 for photodynamic therapy. AB - BACKGROUND: Photodynamic therapy (PDT) has been shown to be effective in the treatment of malignancies of a variety of organ systems, including the lungs, bladder, gastrointestinal tract and skin. Cutaneous lesions serve as ideal targets of PDT because of the accessibility of the skin to light. To achieve optimum results, the photosensitizer must be delivered effectively into the target layers of the skin within a practical timeframe, via noninvasive methods. AIM: To determine whether topical application of a second-generation photosensitizer, silicon phthalocyanine (Pc) 4 [SiPc(OSi(CH3)2 (CH2)3 N(CH3)2)(OH)], results in effective penetration of the skin barrier. METHODS: Penetration of Pc 4 was evaluated using standard Franz-type vertical diffusion cell experiments on surrogate materials (silicone membranes) and laser-scanning confocal microscopy of normal skin biopsy samples from human volunteers. RESULTS: The Franz diffusion data indicate that Pc 4 formulated in an ethanol/propylene glycol solution (70/30%, v/v) can penetrate the membrane at a flux that is appreciable and relatively invariant. Using the same formulation, Pc 4 uptake could be detected in human skin via laser-scanning confocal microscopy. CONCLUSION: After topical application, Pc 4 is absorbed into the epidermis in as little as 1 h, and the absorption increased with increasing time and dose. Pc 4 can be effectively delivered into human skin via topical application. The data also suggest that the degree of penetration is time- and dose-dependent. PMID- 21623876 TI - Expression of RUNX3 in skin cancers. AB - BACKGROUND: Expression of Runt-related transcription factor 3 (RUNX3) is reduced in a large number of cancers. However, a few studies have reported higher expression of RUNX3 in several cancers, including basal cell carcinoma (BCC). In light of this, we explored the expression of RUNX3 in skin cancers generally, to determine whether it acts as an oncogene or a tumour-suppressor gene in skin tumours. AIM: To investigate the expression of RUNX3 in normal skin and malignant skin tumours. METHODS: RUNX3 expression was evaluated by western blotting in 24 specimens, comprising 6 malignant melanoma (MM), 6 squamous cell carcinoma (SCC), 6 BCC and 6 normal skin specimens. Immunohistochemical staining was carried out to analyse RUNX3 expression in 16 MM, 16 SCC and 16 BCC specimens. To identify where the protein was expressed, the cytoplasmic and nuclear protein expression of RUNX3 in skin cancer tissues was determined. A cell-proliferation study was performed on an MM line (G361) by small interfering (si)RNA transfection. RESULTS: The western blotting experiments showed that RUNX3 was not expressed in normal skin tissues, but it was overexpressed in all MM and SCC samples, and in five of the six BCC samples. Using immunochemistry, RUNX3 was found to be overexpressed in all cancer tissues analysed. Subcellular fraction analysis revealed that RUNX3 was expressed in the nuclei but not the cytoplasm of all the skin cancer tissues analysed, and RUNX3 silencing by siRNA in G361 cells resulted in a decrease in proliferation. CONCLUSIONS: Based on these results, we suggest that RUNX3 has an oncogenic potential and does not act as a tumour suppressor in skin cancers. PMID- 21623877 TI - High levels of LIGHT and low levels of soluble herpesvirus entry mediator in sera of patients with atopic dermatitis. PMID- 21623878 TI - A rationale for systemic treatment in onychomycosis with negative results on fungal examination. AB - BACKGROUND: Fungal infection of the nail affects millions of people worldwide, and has an estimated prevalence of about 10% of the general population. Laboratory confirmation of fungal infection is currently accepted as a requirement before initiation of antifungal treatment in clinical practice. AIM: To examine the rationale for systemic treatment in cases of clinical onychomycosis with negative results on fungal examination (potassium hydroxide test and fungal culture). METHODS: In total, 147 patients with suspected clinical toenail onychomycosis but with negative results on fungal examination underwent up to three consecutive fungal examinations of the affected nails. Patients who were negative after these examinations underwent a fourth set of investigations, including PCR. RESULTS: Of the 147 cases initially thought to be negative, 138 (94%) were rated as positive after up to four consecutive sets of laboratory mycological investigations including PCR. Trichophyton rubrum was by far the commonest dermatophyte cultured from all samples. CONCLUSIONS: In the majority of cases of initially negative examinations, consecutive laboratory fungal tests will eventually produce positive results. These findings suggest that systemic antifungal treatment should be started in patients with suspected fungal infections, even if they have negative laboratory fungal examinations. PMID- 21623879 TI - Accumulation of S-100+ CD1a+ Langerhans cells as a characteristic of lichen nitidus. PMID- 21623880 TI - Novel clinical and molecular findings in Chinese families with Hailey-Hailey disease. PMID- 21623881 TI - Annular elastolytic giant cell granuloma associated with diabetes mellitus: a case report and review of the Japanese literature. PMID- 21623882 TI - Renbok phenomenon between psoriasis and alopecia areata. PMID- 21623883 TI - Botulinum toxin type A in the management of a neuropathic foot ulcer. PMID- 21623884 TI - Chronic granulomatous disease of childhood: an unusual cause of recurrent uncommon infections in a 61-year-old man. AB - Chronic granulomatous disease (CGD) is a rare congenital immunodeficiency that affects 1 : 250,000 of the population, which is characterized by recurrent bacterial and fungal infections and by granuloma formation. We investigated a 61 year-old man presented with a 20-year history of a relapsing skin rash appearing as mildly pruritic and erythematous plaques affecting various body regions. Cutaneous biopsies were taken and sent for histology and tissue culture. Leucocyte function was assessed by determining the generation of reactive oxygen species. Bactericidal activity was assessed in the presence of autologous and homologous sera. Western blotting was performed for protein analysis of the reduced nicotinamide adenine dinucleotide phosphate oxidase system, and mutation screening was carried out using PCR amplification and sequence analysis. Examination of biopsies obtained from lesional skin indicated a suppurative granulomatous process. Tissue cultures grew Aspergillus nidulans and Aspergillus fumigatus (confirmed by PCR). A. nidulans has often been associated with CGD, and the leucocyte function tests supported this diagnosis. Direct DNA sequencing led to the identification of a hemizygous missense novel mutation in CYBB (c.907C>T), which predicts a p.His303Tyr amino-acid substitution in gp91-phox, thus confirming the diagnosis of CGD. In conclusion, we report a case of a rare inherited immunodeficiency, CGD, in a 61-year-old man, and describe the novel hemizygous missense mutation underlying the condition. Mild forms of usually fatal immunodeficiencies should be considered when assessing the occurrence of unusual infectious diseases in apparently healthy people. PMID- 21623885 TI - Drug-induced bullous pemphigoid: cases triggered by intravenous iodine and etanercept. AB - Bullous pemphigoid (BP) is one of the most common acquired immunobullous diseases. Rarely, the development of BP is attributed to drug exposure. We present two cases of BP, one triggered by intravenous iodine, and one associated with etanercept treatment in a patient with psoriasis; the first time, to our knowledge, that either of these associations has been reported. The recognition of occasional cases of drug-induced BP such as ours, with timely cessation of the offending agent, may produce rapid clinical improvement with decreased exposure to potent immunosuppressive therapy. PMID- 21623886 TI - The alpha-melanocyte-stimulating hormone-melanocortin receptor system influences the effects of ultraviolet A on skin and intestinal immunity in mice. AB - Irradiation by ultraviolet (UV) A is known to decrease Langerhans cells (LCs) in the skin and increase IgA expression in the intestine, specifically the jejunum. These changes were induced in C57BL/6j mice by exposure of the ear or the eyes to 11 J/cm2 UVA radiation, then a melanocortin receptor agonist (Agouti-related protein; AgRP) was introduced either intracerebrally or intracerebroventricularly. The degree of change in both LC number and IgA expression induced by UVA eye irradiation was reduced more by intracerebral than by intraperitoneal injection of AgRP. alpha-Melanocyte-stimulating hormone and melanocortin receptors in the brain seem to contribute to immunomodulation after UVA irradiation of the eye or the ear in mice. PMID- 21623887 TI - Acquired progressive kinking of hair affecting the scalp and eyelashes in an adult woman. AB - Acquired hair kinking of scalp hair in adults is an unusual, rarely reported phenomenon. There are several possible aetiologies, including drug-induced changes, diffuse partial woolly hair, and acquired progressive kinking of hair (APKH). We report a case of focal APKH occurring in an adult woman. This was associated with a previously unreported feature, that of eyelash involvement. PMID- 21623888 TI - Post-flooding disaster crop diversity recovery: a case study of Cowpea in Mozambique. AB - To restore food security to a traditional African cropping system following a sudden loss of seed, genetic diversity must be re-established. This study examines the extent to which Cowpea diversity was reinstated two years after a flood disaster in Gaza Province, Mozambique. The contribution that seed from various sources made to the recovery was assessed using semi-structured interviews and morphological and molecular data. Data suggest that diversity had recovered to some extent yet there was evidence of a narrowing of the genetic base, with fewer rare alleles and differences in the distribution of allele frequencies. Although the main channels for accessing seed after the flood were seed relief and markets, these sources contributed to minimal and different diversity. It appears that diversity was regained primarily through social networking in the form of loans or gifts of seed from friends and relatives. The results of the study are discussed in relation to seed relief approaches. PMID- 21623889 TI - Latino social network dynamics and the Hurricane Katrina disaster. AB - The aim of this qualitative research was to examine the dynamics of existing and emerging social networks among Latino survivors of Hurricane Katrina. Data were generated through individual, in-depth interviews conducted with 65 Latinos within six months of the storm striking the Gulf Coast of the United States in August 2005. The findings illustrated both the role of social networks in gathering information, making decisions and accessing resources, and how these existing social networks were disrupted and strained by overwhelming needs. Broader structural issues, including poverty and a lack of transportation, combined with marginalised status as immigrants, further constrained access to essential information and resources. In response, new, if temporary, social networks emerged, based primarily on shared nationality, language, and a sense of collective commitment. Practice implications include the need to consider the social network dynamics of marginalised groups in developing innovative strategies to overcome structural barriers to accessing resources essential for disaster preparedness and survival. PMID- 21623890 TI - Controlling disasters: recognising latent goals after Hurricane Katrina. AB - Classic sociological theory can be used to interpret the aftermath of Hurricane Katrina, which made landfall in the United States on 29 August 2005. The delayed and ineffective response to the storm and the subsequent failure of the levees become more understandable when one considers the latent goals of social control in disaster recovery. Constructing the survivors as suspect or criminal and conceptualising the impacts of the disaster as individual problems occurred in order to justify the emphasis on controlling the survivors of Katrina rather than on assisting them. Parallels are drawn here between the disaster response featuring social control efforts and a recent paradigmatic shift in criminal justice from justice to 'risk management'. Recognition of the implicit aims of the inadequate disaster response provides a more complete explanation of why post Katrina efforts failed to achieve the manifest goals of response and recovery. The conclusion suggests ways to ensure more equitable and just disaster responses. PMID- 21623891 TI - Preventing corruption in humanitarian assistance: perceptions, gaps and challenges. AB - Corruption is a threat to the purpose of humanitarian assistance. Until fairly recently, humanitarian assistance has not been considered an important arena in broader efforts aimed at curbing corruption, and corruption has not always been considered a particularly important concern for humanitarian assistance despite the obviously challenging nature of the context of humanitarian emergencies. Corruption, though, is a threat to humanitarian action because it can prevent assistance from getting to the people who most need it, and because it can potentially undermine public support for such assistance. This paper examines perceptions of corruption and its affects, documents best practices, and outlines gaps in understanding. It suggests recommendations for improving the capacity of humanitarian agencies to prevent and manage the risk of corruption. Agencies have taken steps to combat corruption and improve accountability--downwards and upwards--but scope remains for improvement and for greater sharing of learning and good practice. PMID- 21623892 TI - The need for innovative strategies to improve immunisation services in rural Zimbabwe. AB - Gokwe South, a rural district in Midlands Province, Zimbabwe, reported the lowest rate of immunisation coverage in the country in 2005: 55 per cent of children vaccinated with three doses of diphtheria/pertussis/tetanus vaccine (DPT3) and 35 per cent dropout between the first and third dose of DPT. In January 2007, the authors assessed local barriers to immunisation and proposed strategies to improve immunisation rates in the district, in the face of nationwide economic and political challenges. A situational analysis was performed to assess barriers to immunisation using focus-group discussions with health workers, key informant interviews with health management and community leaders, and desk reviews of records. Responses were categorised and solutions proposed. Health workers and key informants reported that immunisation service delivery was hampered by insufficient availability of gas for cold-chain equipment, limited transport and fuel to conduct basic activities, and inadequate staff and supervision. Improving coverage will require prioritising gas for vaccine cold-chain equipment, identifying reliable transportation or alternative transportation solutions, and increased staff, training and supervision. Local assessment is critical to pinpointing site-specific barriers, and innovative strategies are needed to overcome existing contextual challenges. PMID- 21623893 TI - Regulation of the cyclopropane synthase cfaB gene in Pseudomonas putida KT2440. AB - In Pseudomonas putida, as in many other eubacteria, cyclopropane fatty acids (CFAs) accumulate in the membrane during the stationary phase of growth. Here, we show that cfaB gene expression in P. putida KT2440 is dependent on the RpoS sigma factor that recognizes the sequence 5'-CTACTCT-3' between -8 and -14. We have carried out a mutational study of the cfa promoter and have determined that positions -9, -12, -13 and -14 are the most critical for maximal activity. In P. putida, the substrates of the CFA synthase, cis-unsaturated fatty acids (cis UFAs), are also substrates for another stress-related enzyme, the cis-trans isomerase (CTI). Despite using the same substrates, we have found that the activity of the CTI is not limited by the CFA synthase activity and vice versa. For instance, in a cfaB knockout mutant, the amount of trans-UFAs synthesized after a specific stress was no higher than in the parental background despite the fact that there are more cis-UFAs available to be used by the CTI as substrates. In this regard, in a cti-deficient mutant background, the levels of CFAs were similar to those in the parental one under the same conditions. PMID- 21623894 TI - Identification of SpiA that interacts with Corynebacterium glutamicum WhcA using a two-hybrid system. AB - The Corynebacterium glutamicum whcA gene is known to play a negative role in the expression of genes responding to oxidative stress. The encoded protein contains conserved cysteines, which likely coordinate the redox-sensitive Fe-S cluster. To identify proteins which may interact with WhcA, we employed a two-hybrid system utilizing WhcA as 'bait'. Upon screening, several partner proteins were isolated from the C. glutamicum genomic library. Sequencing analysis of the isolated clones revealed out-of-frame peptide sequences, one of which showed high sequence homology with a dioxygenase encoded by NCgl0899. In vivo analysis of protein interaction using real-time quantitative PCR, which monitors his3 reporter gene expression, demonstrated that the interaction between NCgl0899-encoded protein and WhcA was specific. The interaction was labile to oxidants, such as diamide and menadione. Based on these data, NCgl0899 was named spiA (stress protein interacting with WhcA). Physical association and dissociation of the purified His(6)-WhcA and GST-SpiA fusion proteins, as assayed by in vitro pull-down experiments, were consistent with in vivo results. These data indicated that the interaction between WhcA and SpiA is not only specific but also modulated by the redox status of the cell and the functionality of the WhcA protein is probably modulated by the SpiA protein. PMID- 21623895 TI - Culturable bacterial community analysis in the root domains of two varieties of tree peony (Paeonia ostii). AB - A total of 985 bacterial strains with different colony characteristics were isolated from the root of tree peony plants (variety 'Fengdan' and 'Lan Furong'); 69 operational taxonomic units were identified by amplified ribosomal DNA restriction analysis. Representatives of each group were selected for partial 16S rRNA gene sequencing and phylogenetic analysis. The major groups in the bulk soil, rhizosphere, and rhizoplane of Fengdan were Firmicutes (63.2%), Actinobacteria (36.3%), and Betaproteobacteria (53.0%), respectively. The major bacteria groups in the bulk soil, rhizosphere, and rhizoplane of Lan Furong were Actinobacteria (34.8%), Gammaproteobacteria (45.2%), and Betaproteobacteria (49.1%), respectively. In total, the bacterial isolates comprised 26 genera--14 in the bulk soil, 14 in the rhizosphere, and 11 in the rhizoplane. The most common genus in the bulk soil of Fengdan and Lan Furong was Bacillus (49.6% and 32.6%, respectively), in the rhizosphere Microbacterium (21.1%) and Pseudomonas (42.0%), and in the rhizoplane Variovorax (53.0% and 49.1%, respectively). The results show that there are obvious differences in the bacterial communities in the three root domains of the two varieties, and the plants exerted selective pressures on their associated bacterial populations. The host genotypes also influenced the distribution pattern of the bacterial community. PMID- 21623896 TI - Examining the genetic variation of reference microbial cultures used within food and environmental laboratories using fluorescent amplified fragment length polymorphism analysis. AB - Fluorescent amplified fragment length polymorphism (FAFLP) analysis was applied to genetically fingerprint 'working culture control strains' used by accredited food microbiology laboratories. A working culture control strain is defined as a subculture from a strain initially obtained from an authenticated source [such as the National Collection of Type Cultures (NCTC)] that is maintained for use with routine testing within the laboratory. Working culture control strains from eight food examination laboratories, representing four bacterial species, were analysed by FAFLP; these were Salmonella Nottingham, Staphylococcus aureus, Listeria monocytogenes and Bacillus cereus. The resultant FAFLP profiles of the eight working culture control strains for each of these species were compared against the appropriate freeze-dried ampoules obtained directly from NCTC. FAFLP results demonstrated that within 50% of working cultures analysed, several laboratories were routinely using working cultures that were genetically different from the original reference NCTC strains. This study highlights the need for laboratories to review the protocols used to process and maintain control strains and working cultures, with a potential view to utilize single-use quality control materials. PMID- 21623897 TI - Purification and characterization of a soluble cytochrome c capable of delivering electrons to chlorate reductase in Ideonella dechloratans. AB - The electron donor for periplasmic chlorate reductase of Ideonella dechloratans has been suggested to be a soluble cytochrome c. We describe here the purification of the 9-kDa periplasmic cytochrome c, denoted cytochrome c-Id1, and demonstrate its ability to serve as an electron donor for purified chlorate reductase. The reaction rate was found to be linearly dependent on the cytochrome c concentration in the range of 0.6-4 MUM. A route for electron transport involving a soluble cytochrome c is similar to that found for other periplasmic oxidoreductases of the dimethyl sulfoxide reductase family, but different from that suggested for the (per)chlorate reductase of Dechloromonas species. PMID- 21623898 TI - Lupus miliaris disseminatus faciei successfully treated with tranilast: report of two cases. AB - We report two cases of lupus miliaris disseminatus faciei (LMDF) in which oral tranilast was effective. In case 1, the patient was a 33-year-old woman who had developed pale red papules on her face, especially around her eyes and lower jaw, approximately 7 months previously. Examination of a skin biopsy specimen revealed epithelioid cell granulomas accompanied by caseous necrosis, and a diagnosis of LMDF was made. The patient was treated successively with azithromycin, roxithromycin and minocycline hydrochloride, but there was no improvement. When we tried oral tranilast therapy, flattening of the papules was observed 2 weeks after the start of treatment, and by 1 month the papules had almost disappeared. In case 2, the patient was a 39-year-old man who had broken out in erythematous papules on both upper and lower eyelids, with some accompanied by scaling, 2 years before the initial examination. Pathological specimen revealed epithelioid cell granulomas accompanied by caseous necrosis, and a diagnosis of LMDF was made. There was no improvement when treated orally with minocycline hydrochloride or doxycycline hydrochloride, and treatment was switched to oral tranilast therapy. After 1 month of treatment, the papules had almost disappeared. We concluded that oral tranilast therapy should be tried as a treatment for intractable LMDF. PMID- 21623899 TI - Detection and identification of probable endemic fungal pathogen, Cryptococcus gattii, and worldwide pathogen, Cryptococcus neoformans, by real-time PCR. AB - A real-time PCR method for detection and identification of Cryptococcus neoformans and Cryptococcus gattii was developed and evaluated using DNA from single-colony or koala nasal smears. Two TaqMan minor groove binder probes that distinguished between these species were designed corresponding to the internal sequences of the CAP59 gene for both species. The real-time PCR assay had 100% specificity, as assessed using 13 reference strains and 300 environmental strains. Twelve smear samples from healthy koalas were analyzed by direct real time PCR. This method successfully detected C. gattii and C. neoformans in one and three koalas, respectively. PMID- 21623900 TI - Comparisons of computed tomography, contrast enhanced computed tomography and standing low-field magnetic resonance imaging in horses with lameness localised to the foot. Part 1: anatomic visualisation scores. AB - REASONS FOR PERFORMING STUDY: To date, few reports exist comparing magnetic resonance imaging (MRI) and computed tomography (CT) for imaging of the equine distal limb, yet clinicians are required to decide which modality to use regularly. OBJECTIVES: To report and compare anatomic visualisation scores obtained for CT, contrast enhanced CT (CECT) and standing low-field MRI (LFMRI) in the equine foot. HYPOTHESIS: Anatomic visualisation score discrepancies would exist between CT, CECT and LFMRI. METHODS: Images of 22 lame horses (31 limbs) undergoing both CT and LFMRI of the foot were reviewed. When available, CECT images were reviewed. The deep digital flexor tendon (DDFT) was categorised into proximal to distal levels (A-D), structures were assigned visualisation scores (Grades 0-3) and technique comparisons were made using the paired marginal homogeneity test. RESULTS: Computed tomography and LFMRI had similar visibility scores for the navicular bone, middle phalanx, DDFT-B, collateral ligaments of the distal interphalangeal joint and collateral sesamoidean ligament of the navicular bone. The proximal and distal phalanx had lower visibility scores with LFMRI. The distal DDFT (C-D), distal sesamoidean impar ligament and synovial structures had higher scores with LFMRI. Contrast enhanced CT lowered DDFT and collateral sesamoidean ligament scores and raised distal interphalangeal synovium CT visualisation scores. CONCLUSIONS AND POTENTIAL RELEVANCE: Visualisation scores differ depending on imaging technique and anatomic structure of interest. This information increases our understanding of the limitations of CT, CECT and LFMRI to visualise anatomy in clinical cases. PMID- 21623901 TI - Positive interaction between lapatinib and capecitabine in human breast cancer models: study of molecular determinants. AB - The combination of lapatinib and capecitabine is approved in Her2+ metastatic breast cancer. However, the pharmacological mechanisms for this association have not been fully elucidated. In this non-clinical study, we evaluated the efficacy of this association on a panel of six human breast cancer cell lines as a means to identify the molecular determinants of response to this combination. Cell viability was evaluated after concomitant/sequential exposure, and response/resistance determinants for each drug such as dihydropyrimidine dehydrogenase (DPD), thymidylate synthase (TS), thymidine phosphorylase, Bax, Bcl2, P21 levels, and phospho p42/44 and HER1/2 signaling pathway were studied. Lapatinib proved to markedly downregulate TS activity, thus suggesting a subsequent better efficacy of capecitabine. Capecitabine optimized the downregulation of p-AKT and p-P42/44 expression by lapatinib. Consequently, we observed an increase in the Bax/Bcl2 ratio and p21 protein expression in cells exposed to the combination. Overall, our data showed that whatever the schedule and the cell line were, additive to synergistic interaction was achieved in our models. The optimal in vitro combination was finally tested in tumor-bearing mice. Our results fully confirmed that associating both drugs led to a 77% reduction in tumor growth as compared with control animals in BT474-xenografted models. Taken together, this non-clinical study shows that lapatinib and capecitabine modulate each other's molecular determinants of response and that concomitant dosing seems to be the optimal way to combine these drugs. Besides, modulation of TS expression by lapatinib makes its association with capecitabine a promising way to overcome breast cancers resistant in relation with TS overexpression. PMID- 21623902 TI - QRS widening and QT prolongation under bupropion: a unique cardiac electrophysiological profile. AB - QRS widening and QT prolongation are associated with bupropion. The objectives were to elucidate its cardiac electrophysiological properties. Patch-clamp technique was used to assess the I(Kr) -, I(Ks) -, and I(Na) -blocking effects of bupropion. Langendorff retroperfusion technique on isolated guinea-pig hearts was used to evaluate the MAPD(90) -, MAP amplitude-, phase 0 dV/dt-, and ECG modulating effects of bupropion and of two gap junction intercellular communication inhibitors: glycyrrhetinic acid and heptanol. To evaluate their effects on cardiac intercellular communication, fluorescence recovery after photobleaching (FRAP) technique was used. Bupropion is an I(Kr) blocker. IC(50) was estimated at 34 MUm. In contrast, bupropion had hardly any effect on I(Ks) and I(Na) . Bupropion had no significant MAPD(90) -modulating effect. However, as glycyrrhetinic acid and heptanol, bupropion caused important reductions in MAP amplitude and phase 0 dV/dt. A modest but significant QRS-widening effect of bupropion was also observed. FRAP experiments confirmed that bupropion inhibits gap junctional intercellular communication. QT prolongation during bupropion overdosage is due to its I(Kr) -blocking effect. QRS widening with bupropion is not related to cardiac sodium channel block. Bupropion rather mimics the QRS widening, MAP amplitude- and phase 0 dV/dt -reducing effect of glycyrrhetinic acid and heptanol. Unlike class I anti-arrhythmics, bupropion causes cardiac conduction disturbances by reducing cardiac intercellular coupling. PMID- 21623903 TI - Increase in invasive nonvaccine pneumococcal serotypes at two hospitals in Barcelona: was replacement disease to blame? AB - AIM: To describe an increase in the incidence of invasive pneumococcal disease (IPD) caused by serotypes not contained in the heptavalent pneumococcal conjugate vaccine (PCV7) in children in two hospitals in Barcelona with different vaccine uptake. METHODS: Cumulative incidences of IPD, vaccine and nonvaccine serotypes (NVSTs), and main clinical presentations before (1998-2001) and after vaccine introduction (2005-2008) were compared. RESULTS: The incidence of IPD in children aged <2 years at Hospital Germans Trias i Pujol covering a population in which PCV7 was not widely used showed a nonsignificant increase from 29.9 to 58.8 per 100,000 child-years between both periods. Following vaccine introduction, there was a 2.5-fold increase in IPD caused by NVSTs in children aged <5 years. Analysis of trends in the almost fully vaccinated population of Hospital de Barcelona revealed a nonsignificant reduction in IPD incidence in children aged <2 years from 63.1 to 26.0 per 100,000 child-years. NVSTs in children aged <5 years showed a nonsignificant 1.7-fold increase in the vaccine period at this centre. CONCLUSIONS: The paradoxical increase in invasive infections caused by NVSTs in these populations with different vaccine use suggests that these changes were not driven only by PCV7. PMID- 21623905 TI - Clinical Transplantation. 14th Japanese Clinico-Pathological Conference on Renal Allograft Pathology. Introduction. PMID- 21623906 TI - Current problems of chronic active antibody-mediated rejection. AB - The Banff 2007 classification allows chronic rejection to be differentiated based on clinicopathological characteristics evidenced by two independent immunologic mechanisms; chronic active antibody-mediated rejection and chronic active T lymphocyte mediated rejection. However, several incompletely understood issues concerning chronic active antibody-mediated rejection remain. Chronic active antibody-mediated rejection is characterized by C4d deposition in the capillary basement membrane(PTC), the presence of circulating anti-donor antibodies(DSA), and morphologic evidence of chronic tissue injury such as glomerular double contours compatible with transplant glomerulopathy (TPG), PTC basement membrane multilayering, interstitial fibrosis/tubular atrophy, and fibrous arterial intimal thickening. PTC basement membrane multilayering correlates highly with TPG, and most of TPG have evidence of either C4d-positive staining or DSA. However, the proposed criteria do not apply to all situations of chronic active antibody-mediated rejection. C4d is not a magic marker for antibody-mediated rejection. C4d staining is not always highly sensitive for detecting antibody mediated rejection. Multi-institutional studies should be conducted to better understand the clinicopathological context of chronic antibody-mediated rejection. These studies should include well-designed serial protocol biopsies with evaluation by electron microscopy, C4d staining performed on frozen sections, and assessment using sensitive DSA detection methods. PMID- 21623907 TI - Focal segmental glomerulosclerosis after renal transplantation. AB - Focal segmental glomerulosclerosis (FSGS) is a clinicopathologic syndrome of proteinuria, usually of nephrotic range, associated with focal and segmental sclerotic glomerular lesions. Therefore, FSGS is diagnosed by clinical features and histopathological examination of renal biopsy. The natural history of the condition varies, and although it may respond to treatment, FSGS is an important disease in the etiology of end-stage renal disease (ESRD). Furthermore, after kidney transplantation, approximately 30% of patients with FSGS develop recurrent FSGS. The risk factors for recurrence of FSGS include childhood onset and age <15 yr, rapid progression of the primary FSGS to ESRD, recurrence of FSGS in a previous allograft, diffuse mesangial hypercellularity in the native kidney, collapsing FSGS, and podocin gene mutation. In addition, after kidney transplantation, de novo FSGS also develops in approximately 10-20% of allografts, associated with a complication of hyperfiltration injury, chronic transplant glomerulopathy, and calcineurin inhibitor toxicity. FSGS is considered a podocyte disease, and the pathology is characterized by segmental FSGS lesion with glomerular epithelial hypercellularity. The pathological diagnosis of FSGS is based on the 2004 Columbia classification system. In the present minireview, we discuss the pathology of recurrence and de novo FSGS after kidney transplantation. PMID- 21623908 TI - A case of tacrolimus-associated thrombotic microangiopathy after ABO-blood-type incompatible renal transplantation. AB - De novo thrombotic microangiopathy(TMA) is most commonly triggered by calcineurin inhibitors (CNI) and the prognosis is less severe than with recurrent TMA. However, it is difficult to distinguish de novo TMA from CNI toxicity and acute antibody-mediated rejection(AMR) soon after renal transplantation. We present a case of tacrolimus-associated TMA soon after ABO blood type incompatible renal transplantation that was difficult to differentiate from acute AMR. On day 9 his urine output decreased dramatically and the Scr level increased. His anti-blood type A antibody titer increased to *16 postopratively and the tacrolimus trough level was higher than in our immunosuppressive regimen. Although we gave priority to anti-AMR treatment, adequate dose adjustment of tacrolimus after tacrolimus nephrotoxicity was diagnosed from graft biopsy could correct allograft dysfunction. PMID- 21623909 TI - A case of progressive thrombotic microangiopathy after ABO-incompatible renal transplantation. AB - A 21-yr-old man of blood type O receiving hemodialysis for IgA nephropathy underwent living-related ABO-incompatible (ABOI) renal transplantation from his mother, whose blood type is A. He was negative for flow cross-match, anti-human leukocyte antigen (HLA) antibody, and anti-MICA antibody. Pre-treatment anti-A IgG titer was 1:256. Desensitization consisted of tacrolimus, mycophenolate mofetil, methylprednisolone, rituximab, and plasmapheresis. He developed acute antibody rejection at day 2 post-transplant, which was successfully treated. After renal artery reconstruction surgery at day 91 for renovascular hypertension caused by renal artery stricture, the patient suffered from acute prostatitis, which subsequently induced type III acute antibody-mediated rejection. Even after recovery from the rejection after temporary hemodialysis, graft function progressively deteriorated and consecutive allograft biopsy showed progressive thrombotic microangiopathy (TMA) without any evidence of donor-specific antibody other than anti-A antibody. The tacrolimus dose was kept low for fear of tacrolimus-induced TMA. Despite these efforts, the patient resumed hemodialysis six months' post-transplant. PMID- 21623910 TI - Is arteriolar vacuolization a predictor of calcineurin inhibitor nephrotoxicity? AB - Calcineurin inhibitors (CNI) have been commonly used as pivotal immunosuppressive agents to renal transplant recipients and have contributed significantly to improving short-term allograft survival. However, long-term administration of CNI may cause an adverse effect on kidney function, known as chronic nephrotoxicity. Chronic CNI nephrotoxicity (CNI-NT) shows characteristic histopathological findings that involve arteriolar hyalinosis. Recently, the term alternative arteriolar hyalinosis (aah) is used to discriminate CNI-specific arteriolar hyaline deposition from non-specific arteriolar hyalinosis. We studied whether arteriolar vacuolization represents an early lesion of aah as a predictor of CNI NT. We retrospectively studied the 79 patients under treatment with a CNI immunosuppressant, who underwent living-related renal transplantation (RTx) from January 2007 to March 2009. We examined serial protocol graft biopsies at one h, one, six, and 12 months after RTx. We classified histological findings into two groups on the basis of aah lesion (with or without aah) in serial biopsies for 12 months. Arteriolar vacuolization was more frequently observed in the aah group than in the non-aah group with a significant difference. Arteriolar vacuolization was found even in the one-h biopsy specimens, indicating a non-specific histopathological finding. But in the aah group, arteriolar vacuolization tended to be more frequently observed later on. Aah can be a predictor of CNI-NT. PMID- 21623911 TI - Successful treatment of nephrotic syndrome caused by recurrent IgA nephropathy with chronic active antibody-mediated rejection three years after kidney transplantation. AB - Here, we report the successful treatment of a 38-yr-old Japanese man diagnosed with recurrent immunoglobulin A nephropathy (IgAN) with chronic active antibody mediated rejection (CAAMR), three yr after undergoing living-related donor kidney transplantation. Immediately after transplantation, the allograft function was well maintained with a serum creatinine (S-Cr) level of <1.8 mg/dL. About three yr after transplantation, urine protein excretion had reached 4.59 g/d, and the S Cr level had increased to more than 2.0 mg/dL. Based on the allograft biopsy, we diagnosed nephrotic syndrome because of recurrence of IgAN with CAAMR. Subsequently, we performed a tonsillectomy, administered three sessions of steroid pulse therapy, and added losartan for the recurrence of IgAN. We also changed his immunosuppressant from mizoribine to mycophenolate mofetil to treat the CAAMR. The nephrotic syndrome improved with the multiple therapeutic approaches; however, the S-Cr level did not decrease below 2.0 mg/dL. We possibly could have performed additional treatments such as rituximab and intravenous immunoglobulin for the CAAMR, but therapeutic strategies for CAAMR have not yet been established. PMID- 21623912 TI - A case of transplant glomerulopathy early after kidney transplantation. AB - Transplant glomerulopathy (TG) has commonly been described as a late manifestation of allograft injury, occurring years after renal transplantation. We describe herein a patient who developed TG early (49 days) after kidney transplantation (KTx). CASE REPORT: A 44-yr-old woman received a second living related KTx from her younger brother in October 2009. Pre-transplant donor specific anti-human leukocyte antigen antibodies (DSA) were positive for both class I and class II. During the renal transplant operation, she suffered from hyperacute antibody-mediated rejection (AMR) and intravenous immune globulin therapy was immediately performed. Once hyperacute AMR was resolved, accelerated acute AMR occurred on the first post-transplant day (PTD). The renal allograft biopsy performed on PTD 19 diagnosed acute AMR type II. The renal allograft biopsy performed on PTD 49 showed focal lesions of double contours of glomerular basement membranes in some glomeruli. Interstitial fibrosis showed a strong, diffuse staining of C4d in peritubular capillaries (PTCs). The DSA examined on PTD 39 were still positive for both class I and class II. From these histopathological findings of TG, C4d deposition in PTC and presence of circulating DSA, we diagnosed this case to have c-AMR. CONCLUSIONS: TG might be recognized in early after KTx. PMID- 21623913 TI - BK polyomavirus nephropathy complicated with acute T-cell-mediated rejection in a kidney transplant recipient: a case report. AB - We present a case of severe BK polyomavirus nephropathy (BKVN) complicated with persistent acute T-cell-mediated rejection (ATMR) that progressed to allograft failure. A 54-yr-old man received a living donor kidney transplant from his wife. Approximately four months after transplantation, the patient's serum creatinine (SCr) increased from a baseline value of 1.5-2.4 mg/dL. A histological analysis showed BKVN, and the SV40 antigen was detected in the tubular nuclei. The doses of immunosuppressants were reduced, and immunoglobulin was administered intravenously. The SCr increased further, to 5.3 mg/dL, and a second renal biopsy revealed the presence of severe ATMR. Antirejection treatment was performed, and low-dose cidofovir was started. The SCr decreased, to 3 mg/dL, and BK virus antigen in the serum and urine samples became negative at the time of hospital discharge. However, the histological findings subsequently showed gradually progressive interstitial fibrosis and tubular atrophy, and the SCr increased gradually. Two years after the transplantation, the patient resumed hemodialysis. BK polyomavirus nephropathy is usually treated with a reduction in immunosuppressant therapy, although in some patients, the reduction in immunosuppressants induces a subsequent exacerbation of acute rejection and results in progressive graft failure, which suggests difficulty in treating BKVN after kidney transplantation. PMID- 21623914 TI - The case of BK virus infection in which it was difficult to differentiate from acute rejection. AB - BK virus (BKV) nephropathy is one of the major causes of allograft dysfunction or graft loss in kidney transplant recipients. Early diagnosis and timely reduction in immunosuppressant is important for proper treatment. We report a 35-yr-old male case of cadaveric renal transplantation with BK viral related tubulointerstitial nephritis complicated by acute rejection. The diagnostic biopsy showed severe inflammatory infiltrates, tubulitis, and peritubular capillaritis. Discontinuation of mycophenolate mofetil, prednisone pulse therapy, and r-globulin was successful in relieving allograft dysfunction. PMID- 21623915 TI - Overlapped glomerular lesions of chronic rejection and recurrent lupus nephritis in transplanted kidney: a case report. AB - We describe a renal transplant recipient with systemic lupus erythematosus (SLE) who showed continuous proteinuria and low complement levels without clinical evidence of active SLE. Her first renal allograft biopsy, performed nine yr and eight months after transplantation, revealed unusual histological change of glomeruli, and it initially led us to make a contradictory diagnosis based on light and electron microscopic examinations. Diffuse global double- or multi contour glomerular basement membrane was caused by chronic endothelial injury owing to chronic rejection, and mesangial proliferation associated with mesangial electron-dense deposit was a histological change characteristic of recurrent lupus nephritis (RLN). Immunofluorescence study displayed weak mesangial staining of IgM and C1q. We concluded that this case presented overlapped chronic rejection and RLN. Because both transplant nephropathy and lupus nephritis present constellations of various histologies, it is difficult to diagnose their overlap. Complete morphologic studies with both immunofluorescence and electron microscopic evaluations in addition to microscopic examination should be performed to elucidate complex histological findings. PMID- 21623916 TI - A case of recurrent focal segmental glomerulosclerosis (FSGS) involving massive proteinuria (>50 g/day) immediately after renal transplantation. AB - A 15-yr-old girl with end-stage kidney disease caused by primary focal segmental glomerulosclerosis (FSGS) underwent a living-related donor kidney transplantation. The allograft functioned well immediately after reperfusion, but massive proteinuria exceeding 50 g/d appeared on day 3. Treatment with rituximab and plasma exchange (PE) successfully decreased the proteinuria to 10 g/d. A biopsy specimen on day 30 showed no segmental glomerulosclerosis but partial interstitial infiltration of inflammatory cells. An increased number of podocytes showed intracytoplasmic vacuolization, and an electron micrograph showed diffuse mild subendothelial edema and foot process effacement. The podocytes were hypertrophied but were not detached from the basement membrane. As the therapies used to reduce the patient's proteinuria were having a limited effect, intravenous steroid pulse therapy followed by low-density lipoprotein apheresis was performed. A biopsy specimen taken on day 120 showed no segmental glomerulosclerosis. Thrombus formation in one glomerulus and packed lymphocytes in the capillary loop of another glomerulus were detected. The patient's clinical course was compatible with FSGS recurrence. Although the early pathological changes were not typical of FSGS, they might be indicative of the primary lesion that subsequently progresses to typical FSGS. PMID- 21623917 TI - Predictors of histology, tissue eosinophilia and mast cell infiltration in Hodgkin's lymphoma--a population-based study. AB - OBJECTIVE: Classical Hodgkin's lymphoma (HL) lesions comprise few tumour cells, surrounded by numerous inflammatory cells. Like in other malignancies, the microenvironment is presumed to be clinically important in HL; however, microenvironment predictors remain poorly characterised. The aim of this study was to investigate how selected patient characteristics and genetic factors affect HL phenotype, in particular tissue eosinophilia, mast cell counts and HL histological subtype. METHODS: In a population-based study, patients with HL were interviewed about potential HL risk factors. Available tumours, n=448, were classified histologically; the number of eosinophils and mast cells were estimated, and eosinophil cationic protein (ECP) and eosinophil protein-x (EPX) gene polymorphisms were determined. Associations were assessed in regression models. RESULTS: Self-reported history of asthma was predictive of having tumour eosinophilia [>=200 eosinophils/10 high power fields, univariate odds ratio (OR)=2.22, 95% CI 1.06-4.64, P=0.03]. High numbers of eosinophils were predominantly seen in patients carrying the genotype ECP434GG [multivariate relative levels (RLs)=1.84, 95% CI 1.02-3.30, P=0.04]. Lower number of eosinophils was seen in Epstein-Barr virus (EBV)-positive tumours (univariate RL=0.52, 95% CI 0.3-0.9, P=0.02) and in older patients (univariate RL=0.85, 95% CI 0.73-0.99, P=0.03). Well-known factors such as young age, female sex and EBV negative status predicted nodular sclerosis histology. CONCLUSION: The number of eosinophils in HL tumours is influenced by patient traits such as asthma, ECP genotype and EBV status. EBV status was predictive of histology. PMID- 21623918 TI - Cytogenetic response is not a prerequisite for clinical response in patients with myelodysplastic syndromes treated with azacitidine. PMID- 21623920 TI - Panniculitis-like T-cell lymphoma detected by positron emission tomography/computed tomography scanning in a patient with haemophagocytic syndrome. PMID- 21623919 TI - Quality of life, physical function and MRI T2* in elderly low-risk MDS patients treated to a haemoglobin level of >=120 g/L with darbepoetin alfa +/- filgrastim or erythrocyte transfusions. AB - OBJECTIVE: Anaemia in low-risk myelodysplastic syndromes (MDS) is associated with reduced quality of life (QoL). Response to treatment with erythropoietin +/- granulocyte colony-stimulating factor (G-CSF) is associated with improved QoL, but whether transfusion therapy with higher haemoglobin (Hb) target levels has similar effects is unknown. The objective for this prospective phase II Nordic multicentre trial was to assess QoL, response rate and physical function in elderly anaemic MDS patients treated to a target Hb level of >120 g/L. METHODS: Thirty-six elderly patients with low- and intermediate-1 risk MDS received darbepoetin (DA) 300 MUg/wk, with the addition of G-CSF if no response. If the Hb target was reached at 16 wk, treatment was maintained until week 26. Remaining patients were transfused to reach the target level for at least 8 wk. RESULTS: Twenty-seven patients completed the study. Response rate to DA +/- G-CSF was 67% in evaluable patients and 56% according to intention to treat. Eighteen patients reached the target Hb level according to protocol. QoL scores for fatigue, dyspnoea, constipation, and physical, role and social functioning improved significantly during study, with similar results for transfused and untransfused patients. Maintaining Hb >120 g/L did not confer a higher transfusion rate, once the target was reached. In two of fourteen patients, magnetic resonance imaging T2* indicated cardiac iron overload, however, without association with ferritin levels. CONCLUSIONS: In elderly anaemic MDS patients, an increment in haemoglobin is associated with improved QoL, whether induced by growth factor treatment or transfusion therapy. PMID- 21623921 TI - Cardiopulmonary bypass in a patient with classic paroxysmal nocturnal hemoglobinuria during treatment with eculizumab. PMID- 21623922 TI - Distribution of dystrophin- and utrophin-associated protein complexes (DAPC/UAPC) in human hematopoietic stem/progenitor cells. AB - Hematopoietic stem cells (HSC) are defined by their cardinal properties, such as sustained proliferation, multilineage differentiation, and self-renewal, which give rise to a hierarchy of progenitor populations with more restricted potential lineage, ultimately leading to the production of all types of mature blood cells. HSC are anchored by cell adhesion molecules to their specific microenvironment, thus regulating their cell cycle, while cell migration is essentially required for seeding the HSC of the fetal bone marrow (BM) during development as well as in adult BM homeostasis. The dystrophin-associated protein complex (DAPC) is a large group of membrane-associated proteins linking the cytoskeleton to the extracellular matrix and exhibiting scaffolding, adhesion, and signaling roles in muscle and non-muscle cells including mature blood cells. Because adhesion and migration are mechanisms that influence the fate of the HSC, we explored the presence and the feasible role of DAPC. In this study, we characterized the pattern expression by immunoblot technique and, by confocal microscopy analysis, the cellular distribution of dystrophin and utrophin gene products, and the dystrophin-associated proteins (alpha-, beta-dystroglycan, alpha-syntrophin, alpha-dystrobrevin) in relation to actin filaments in freshly isolated CD34+ cells from umbilical cord blood. Immunoprecipitation assays demonstrated the presence of Dp71d/Dp71Delta110m ~DAPC and Up400/Up140~DAPC. The subcellular distribution of the two DAPC in actin-based structures suggests their dynamic participation in adhesion and cell migration. In addition, the particular protein pattern expression found in hematopoietic stem/progenitor cells might be indicative of their feasible participation during differentiation. PMID- 21623923 TI - G-CSF enhances the proliferation and mobilization, but not the maturation rate, of murine myeloid cells. AB - OBJECTIVES: Whether G-CSF enhances the maturation of neutrophilic granulocytes or just accelerates the mobilization of mature and maturing granulocytes from bone marrow to blood, or both, is not clear. Using an in vivo culture system where such mobilization cannot take place, we previously showed that G-CSF did not accelerate maturation. To further clarify the role of G-CSF, we now have examined its effect on murine granulopoiesis in situ. METHODS: Murine bone marrow precursors in S-phase were labeled with BrdU, and hematopoiesis stimulated by the long-acting G-CSF compound pegfilgrastim (peg-G-CSF). Performing flow cytometric analysis of incorporated BrdU and the granulocyte maturation antigen Gr1, we investigated the cell flux from the proliferative to the non-proliferative granulocyte compartments in bone marrow and further from bone marrow to blood. RESULTS: Peg-G-CSF mobilized neutrophils from bone marrow to blood and markedly increased their concentration in blood for several days. It also increased the proliferation of precursor cells. Newly produced, less mature granulocytes (Gr1(+) BrdU(+)) travelled faster to blood in treated mice than in controls. The flow cytometric and cell density analyses of the bone marrow cells showed that peg-G-CSF skewed the population toward less mature cells, mainly because of the mobilization of granulocytes to blood. CONCLUSIONS: Collectively, our data do not support the notion that G-CSF accelerates murine granulocyte maturation per se. PMID- 21623924 TI - ORAOV1 is amplified in oral squamous cell carcinoma. AB - BACKGROUND: Oral cancer overexpressed 1 (ORAOV1) was found as a candidate oncogene in the 11q13 chromosomal region, based on its amplification and overexpression in oral cancer cell lines. Because gene amplification often leads to increased levels of gene expression, we aimed to verify the relationship between ORAOV1 gene status and mRNA expression primarily in oral squamous cell carcinoma (OSCC) by quantitative assay, correlating with clinical and pathological characteristics in patients. METHODS: Levels of ORAOV1 amplification and expression were evaluated by qPCR and RT-qPCR in OSCC cell lines and in tumor and non-tumoral surgical margins from 33 patients with OSCC. All subjects were smokers and habitual alcohol drinkers, mostly men above 40 years of age and with a single primary tumor. RESULTS: ORAOV1 exhibited increased gene expression levels as well as higher copy number in three OSCC cell lines with 11q13 amplified chromosomal region when compared with the OSCC cell line without the amplification (one-way ANOVA, P < 0.05). Weak correlation between ORAOV1 mRNA levels and DNA copy number was seen in tumor samples (Spearman, P = 0.07). Although ORAOV1 was amplified in tumor (Wilcoxon, P < 0.01), high levels of transcripts in margin did not reveal differences in comparison with tumor (Wilcoxon, P = 0.85). Aggressiveness and survival rate did not demonstrate statistical difference for both events in OSCC. CONCLUSION: The overexpression of ORAOV1 in non-tumoral margin samples can occur in the absence of amplification. The weak correlation between ORAOV1 amplification and expression in OSSC suggests that ORAOV1 expression can be regulated by mechanisms other than gene amplification. PMID- 21623925 TI - Infliximab partially impairs the anti-Mycobacterium tuberculosis immune responses of severe psoriasis patients with positive tuberculin skin-test. AB - BACKGROUND: Infliximab and etarnecept are now widely used for treating severe psoriasis. However, these drugs, especially infliximab, increased the risk of tuberculosis reactivation. Surprisingly, epidemiological data suggest that the tuberculosis rate in patients taking infliximab in Sao Paulo State, Brazil, is similar to that of some developed, non-endemic countries. OBJECTIVE: The aim of this study was to better understand the effect of infliximab on Mycobacterium tuberculosis (Mtb) immune responses of psoriasis patients in an endemic setting (Brazil). METHODS: We evaluated the tuberculosis-specific immune responses of severe psoriasis patients and healthy individuals, both tuberculin skin test (TST) positive, in the presence/absence of infliximab. Patients had untreated severe psoriasis, no co-morbidities affecting the immune responses and a TST >10 mm. Healthy TST(+) (>10 mm) individuals were evaluated in parallel. PBMC cultures from both groups were stimulated with different Mycobacterium tuberculosis (Mtb) antigens (ESAT-6, 85B and Mtb lysate) and phytohemagglutinin, with or without infliximab (5 MUg/mL). Parameters evaluated were TNF-alpha, IFN-gamma and IL-10 secretion by ELISA, overnight IFN-gamma ELISpot and lymphocyte proliferative response (LPR). RESULTS: Infliximab almost abolished TNF-alpha detection in PBMC supernatants of both groups. It also significantly reduced the LPR to phytohemagglutinin and the Mtb antigens as well as the IFN-gamma levels secreted into day 5 supernatants in both groups. There was no concomitant exaggerated IL 10 secretion that could account for the decreases in these responses. ELISpot showed that, contrasting with the central-memory responses above, infliximab did not affect effector-memory INF-gamma-releasing T-cell numbers. CONCLUSIONS: Infliximab affected some, but not all aspects of the in vitro antituberculosis immune responses tested. The preserved effector-memory responses, putatively related to exposure to environmental mycobacteria, may help to explain the lower than expected susceptibility to tuberculosis reactivation in our setting. PMID- 21623926 TI - Finger retronychias detected early by 3D ultrasound examination. AB - BACKGROUND: A new pattern of ingrown nail, called retronychia is involved in the posterior translation of the whole nail unit producing paronychia. OBJECTIVE: To demonstrate an unusual case of paronychia that affected three of the fingers on the right hand. METHODS: We used 3D ultrasound to study the whole nail unit in the fingers of both the affected (right) and non affected (left) hand. RESULTS: Backward motion of the nail unit with decreased distance between the origin of the nail plates and the distal interphalangeal joint was clearly demonstrated by 3D ultrasound in the affected fingers. CONCLUSION: 3D ultrasound provides non invasive and more understandable information about the physiopathological changes in retronychia. PMID- 21623927 TI - Natural options for the management of hyperpigmentation. AB - Facial hyperpigmented disorders are a common complaint in the adult population of all races. First-line topical treatments are usually hydroquinone or topical retinoids, which can cause irritant reactions. The need for better tolerated, yet effective, skin lightening agents that could be utilized by a wider population has led to the investigation of several potential botanical/natural compounds. There are currently many topical cosmetic formulations claiming skin depigmenting effects. A few of the ingredients (e.g. soy) are supported not only by in vitro results but also by a body of controlled clinical efficacy studies; other ingredients, instead, are backed mostly by in vitro data and a few small uncontrolled clinical studies. In this review, we describe the most common natural ingredients used for skin depigmentation and their major published studies: soy, licorice extracts, kojic acid, arbutin, niacinamide, N acetylglucosamine, COFFEEBERRY(TM) and green tea. PMID- 21623928 TI - Differences between familial and sporadic cases of vitiligo. AB - BACKGROUND: Most cases of vitiligo are sporadic, but about 10-36% of the patients have positive family history. OBJECTIVE: The aim of our study was to describe differences between familial and sporadic cases of vitiligo. METHODS: A total of 186 adult vitiligo patients were examined, in 173 of whom the level of thyroid peroxidase antibodies, gastric parietal cell antibodies (PCA), antinuclear antibodies (ANA), anti-adrenal cortex antibodies and rheumatoid factor in blood was measured. All patients were divided in two groups: the cases with positive family history of vitiligo (51) and the sporadic cases (135). RESULTS: The risk of onset of the disease up to 20 years of age was higher in the familial group (P=0.008). Patients in familial group showed more widespread depigmentation compared with sporadic cases [body surface area (BSA) over 10%: P=0.004; BSA over 50%: P=0.001]. In familial group, patients had darker skin phototype (P=0.045) and the disease had started more often as a vulgar vitiligo (P=0.008). In sporadic vitiligo group, female gender was a risk factor for more widespread depigmentation (BSA over 10%, P=0.001). Extensive depigmentation was associated with reported triggering factors and mucosal involvement in both groups and with leukotrichia only in familial group. Widespread depigmentation related to the risk of presence of autoantibodies (P=0.03) in sporadic cases of vitiligo (especially of PCA: P=0.04 and ANA: P=0.0002). CONCLUSIONS: In this study, we demonstrated first time that patients with familial vitiligo have a higher risk for vulgar type at the beginning of the disease and female gender increases the risk for more extensive depigmentation in sporadic cases. PMID- 21623929 TI - The effect of topically applied corticosteroids on interleukin 1beta levels in patients with atopic dermatitis. AB - BACKGROUND: Atopic dermatitis (AD) is a chronic recurrent disease of childhood. The therapeutics of AD needs to be more tailored given the new cytokine antibodies available. In this scenario we considered interleukin 1 beta (IL 1beta) levels before and after therapy with topical corticosteroids. OBJECTIVES: To measure the IL-1beta levels before and after treatment in patients with AD after 75% improvement in scoring for atopic dermatitis index. METHODS: Forty-five subjects with AD were treated with betamethasone ointment and their IL-1beta level was measured prior to and following treatment. RESULTS: The IL-1beta levels were raised during disease activity and following improvement they decreased in a statistically significant manner. Also the IL-1beta levels corresponded with the severity of disease activity. CONCLUSIONS: Newer modalities of therapy in the form of cytokine monoclonal antibodies should be studied in a larger scale in AD. PMID- 21623930 TI - Preventing melasma recurrence: prescribing a maintenance regimen with an effective triple combination cream based on long-standing clinical severity. AB - BACKGROUND: The relapsing nature of melasma emphasizes the need to maintain efficacy achieved after acute treatment. OBJECTIVE: To compare clinical efficacy and safety of two 6-month Triple Combination (TC; containing fluocinolone acetonide, hydroquinone and tretinoin) maintenance regimens in subjects with moderate to severe melasma, after daily treatment up to 8 weeks. METHODS: This randomized, investigator-blinded, controlled study had a maintenance phase of 6 months. Sixteen centres in Brazil and Mexico enrolled 242 subjects 18 years or older attaining no or mild melasma after 8 weeks of daily TC applications. Subjects were randomized to receive TC in a twice weekly or tapering regimen [3/week (1st month), 2/week (2nd month), 1/week (4th month)]. Efficacy and safety measurements included median time to relapse and relapse-free rate, Global Severity Score, Melasma Area and Severity Index score (MASI), subject's assessment, quality of life questionnaire (MelasQol), and adverse events. RESULTS: The majority (78.8%) had no or mild melasma (GSS <= 1) at week 8 and entered maintenance phase. After 6 months, 53% of patients remained relapse-free with improved quality of life, and time to relapse was similar between groups (about 190 days). Melasma severity at study entry, not maintenance baseline, influenced relapse rate. The twice weekly regimen tended to show better effectiveness in postponing relapse in severe melasma. Both regimens were safe. CONCLUSIONS: After resolution of melasma with TC, maintenance therapy over 6 months was successful in preventing relapse in over half of the patients who entered maintenance phase. Prescribing medicines should be adapted to patients based on melasma severity. PMID- 21623931 TI - Comparative therapeutic efficacy and safety of type-II collagen (UC-II), glucosamine and chondroitin in arthritic dogs: pain evaluation by ground force plate. AB - The investigation was conducted on client-owned moderately arthritic dogs with two objectives: (i) to evaluate therapeutic efficacy of type-II collagen (UC-II) alone or in combination with glucosamine hydrochloride (GLU) and chondroitin sulphate (CHO), and (ii) to determine their tolerability and safety. Dogs in four groups (n = 7-10), were treated daily for a period of 150 days with placebo (Group-I), 10 mg active UC-II (Group-II), 2000 mg GLU + 1600 mg CHO (Group-III), and UC-II + GLU + CHO (Group-IV). On a monthly basis, dogs were evaluated for observational pain (overall pain, pain upon limb manipulation, and pain after physical exertion) using different numeric scales. Pain level was also measured objectively using piezoelectric sensor-based GFP for peak vertical force and impulse area. Dogs were also examined every month for physical, hepatic (ALP, ALT and bilirubin) and renal (BUN and creatinine) functions. Based on observations, significant (p < 0.05) reduction in pain was noted in Group-II, III, and IV dogs. Using GFP, significant increases in peak vertical force (N/kg body wt) and impulse area (N s/kg body wt), indicative of a decrease in arthritis associated pain, were observed in Group-II dogs only. None of the dogs in any group showed changes in physical, hepatic or renal functions. In conclusion, based on GFP data, moderately arthritic dogs treated with UC-II (10 mg) showed a marked reduction in arthritic pain with maximum improvement by day 150. UC-II, GLU and CHO operate through different mechanisms of action, and were well tolerated over a period of 150 days. PMID- 21623932 TI - Modulation of electrogenic transport processes in the porcine proximal colon by enteric neurotransmitters. AB - The aim of our study was to evaluate the involvement of essential pro- and antisecretory neurotransmitters in regulation of secretion in porcine proximal colon. Choline acetyltransferase (ChAT), nitric oxide synthase (NOS), vasoactive intestinal peptide (VIP), substance P (SP), somatostatin (SOM) and neuropeptide Y (NPY) were located immunohistochemically in the epithelium and subepithelial layer. Modulation of epithelial secretion was studied in Ussing chambers. Application of carbachol (CA), sodium nitroprussid (SNP), VIP and SP but not of NPY or SOM resulted in a chloride dependent increase in short circuit current (I(sc) ). I(sc) increase induced by CA, VIP or SNP was not altered by preincubation with tetrodotoxin or indomethacin. In contrast, SP-induced I(sc) increase was diminished by preincubation with tetrodotoxin, indomethacin, L-nitro arginin-methyl-ester, and atropine but not hexamethonium. Simultaneous application of CA and VIP, or CA and SNP increased the I(sc) stronger as expected. Applying SP/CA led to a smaller increase in I(sc) as calculated. It is concluded that mainly prosecretory neurotransmitters are involved in regulation of colonic secretion. Cross-potentiations of acetylcholine and nitric oxide and acetylcholine and VIP suggest activation of different intracellular cascades. Similar intracellular pathways may be stimulated by acetylcholine and SP, thus preventing an additive effect of the transmitters. PMID- 21623933 TI - Observation of micro- and macroconidia in Trichophyton violaceum from a case of Tinea faciei. PMID- 21623934 TI - Late-presenting vascular graft infection caused by Aspergillus in an immunocompetent patient. PMID- 21623935 TI - Discrimination between the enzymatic activities of Candida albicans pleomorphic forms determined using the api(r) ZYM test. AB - Enzymatic activity profiles for two morphotypes of 37 Candida albicans clinical isolates were compared. Yeast and hyphal forms were grown using yeast extract peptone-glucose broth or undiluted human serum, respectively. Both morphotypes were documented under scanning electron microscopy. The api((r)) ZYM (BioMerieux, France) test was used to evaluate the enzymatic activity profiles for particular pleomorphic forms. None of the examined enzymatic activities showed good agreement (kappa, kappa > 0.80) for the two morphotypes of the tested strains. Only leucine arylamidase activity in blastoconidia and hyphae of 35 out of 37 strains appeared to be in significant agreement (kappa = 0.770). This phenomenon should be explored further for clinical benefits. For morphotypes of all tested strains, activity profiles of 11 hydrolytic enzymes demonstrated weak agreement (kappa = 0.044-0.197). Moreover, satisfactory (kappa = 0.218-0.348) and moderate agreement (kappa = 0.413-0.479) were noted for enzymatic activity values of five and two enzymes, respectively. The distinct differences in activity profiles of hydrolytic enzymes between hyphae and blastoconidia is suggested to be related to the specific roles of these two morphotypes in particular steps of pathogenesis. Moreover, both morphotypes should be examined by strain biotyping methods. Beta-N hexosaminidase (HexNAcase) activity assessed by the api((r)) ZYM test and on CHROMagar Candida((r)) medium (Becton Dickinson, USA) is also discussed. PMID- 21623936 TI - Cryptococcuria as manifestation of disseminated cryptococcosis: Staib agar as a selective identification medium. AB - We conducted a retrospective study of 58 cases of cryptococcosis (1986-2008) with urine test positive for Cryptococcus sp, in Mycology Laboratory, Santa Casa Hospital Complex, Porto Alegre, RS, Brazil. The diagnosis of cryptococcuria was based on microscopic examination and culture of urinary sediment. Cryptococcus was isolated from other clinical specimens such as blood, cerebrospinal fluid, ascitic and pleural fluids, respiratory secretions, biopsies of skin, nasal and bone marrow. Cryptocccus neoformans was present in 55 cases and Cryptocccus gattii in three cases. Males predominated (79.3%); age ranged from 12 to 86 years. Acquired Immune Deficiency Syndrome (AIDS) were present in 60.3%, 31.1% did not have AIDS and 5.2% were apparently immunocompetent patients. The most frequent signs and symptoms were headache (53.4%) and fever (51.7%). The most widely used medication was the amphotericin B (43 patients). The mortality rate was 45%. We conclude that the mycological examination of the urine can be an alternative simple, non-invasive and useful in diagnosis of disseminated cryptococcosis, especially when used in conjunction with techniques for demonstration of the capsule (nigrosine) and/or production of melanin in special culture media (Staib agar). PMID- 21623937 TI - Enteral nutrition may cause false-positive results of Aspergillus galactomannan assay in absence of gastrointestinal diseases. PMID- 21623939 TI - Mentorship programmes for laparoscopic and robotic urology. PMID- 21623940 TI - Educational research in urology: current status and future challenges. PMID- 21623941 TI - Survival after prostate brachytherapy in patients aged 60 years and younger. PMID- 21623942 TI - Regional lymph node status in patients with bladder cancer found to be pathological stage T0 at radical cystectomy following systemic chemotherapy. PMID- 21623943 TI - Long-term follow-up of T1 high-grade bladder cancer after intravesical bacille Calmette-Guerin treatment. PMID- 21623946 TI - A novel circumferential bladder neck suture to facilitate vesicourethral anastomosis during radical retropubic prostatectomy. PMID- 21623950 TI - Correlation between phospholipase of Candida albicans and resistance to fluconazole. AB - The objective of this study was to compare phospholipase production between fluconazole-resistant and fluconazole-susceptible strains of Candida albicans in order to explore the relationship between resistance to antifungal drugs and virulence of C. albicans. Fifteen each of fluconazole-resistant (MIC >= 64 MUg ml(-1)) and fluconazole-susceptible (MIC <= 8 MUg ml(-1)) strains of C. albicans were incubated on egg yolk agar to detect phospholipase activity. Virulence of C. albicans was assessed by the average survival time of infected mice. Expression of phospholipase B1 mRNA and protein were detected by RT-PCR and Western blot method. Significant differences between the two groups of Candida strains were observed in phospholipase activity and average survival time of infected mice. The expression of phospholipase B1 mRNA and protein (both of secreted and intracellular forms) were higher in resistant strains than in susceptible strains. The results indicate that the phospholipase activity of C. albicans may be related to its resistance to antifungal drugs. PMID- 21623951 TI - Mucormycosis in paediatric patients: demographics, risk factors and outcome of 12 contemporary cases. AB - Mucormycosis is associated with high morbidity and mortality and is perceived as an emerging fungal infection. However, contemporary paediatric data are limited. We present a series of paediatric cases of mucormycosis reported from Germany and Austria collected within a voluntary epidemiological survey through standardised, anonymized case report forms. Twelve cases were reported between January 2004 and December 2008 (six men; mean age: 12.6 years, range: 0.1-17 years). Mucormycosis was proven in nine, and probable in three cases. Isolates included Lichtheimia (syn. Absidia pro parte, Mycocladus) (five), Rhizopus (three) and Mucor (one) species. Infection was limited to soft tissue in three cases, the lung in two cases, and an infected thrombus in one case; rhinocerebral disease was found in three cases, and pulmonary-mediastinal, pulmonary-cerebral and soft tissue cerebral involvement in one case each. All three patients with isolated soft tissue infection were cured, whereas seven of the remaining patients died (one patient without follow-up). The overall mortality rate was 67%. While these data cannot provide conclusive data on incidence and disease burden of mucormycosis in paediatric patients, they reflect the continuing threat of these infections to immunocompromised patients and the need for improved diagnosis and management. PMID- 21623952 TI - Baculovirus as delivery system for gene transfer during hypothermic organ preservation. AB - Concerns over the safety of conventional viral vectors have limited the translation of gene transfer from an exciting experimental procedure to a successful clinical therapy in transplantation. Baculoviruses are insect viruses, but have the ability to enter mammalian cells and deliver potential therapeutic molecules with no evidence of viral replication. This study provides evidence of the ability of recombinant baculovirus to enter mammalian kidneys and livers during cold preservation. Six kidneys and six liver lobules retrieved from large pigs were perfused with University of Wisconsin (UW) solution containing a baculovirus tagged with green fluorescent protein and preserved for 8 h. In addition, six kidneys were perfused with UW containing a baculovirus expressing red fluorescent protein and preserved for 24 h. Green fluorescent virus particles were detected within transduced kidneys and livers after 8 h standard cold storage and red fluorescent protein mRNA was detected in kidneys after 24 h of cold preservation. There were no significant differences in tissue architecture, cell morphology or ATP content between experimental organs and their controls. Ex vivo transduction of organs with recombinant baculovirus during conventional cold preservation was demonstrated with no evidence of additional injury or reduction in cell viability. PMID- 21623953 TI - The role of interleukin-17 in murine cytomegalovirus interstitial pneumonia in mice with skin transplants. AB - We hypothesized that the T helper (Th)17 response plays an important role in murine cytomegalovirus (MCMV) interstitial pneumonia. BALB/c mice with skin grafts from C57/BJ6 mice were intranasally inoculated with 1.0 * 10(5) PFU MCMV. Lung tissues and skin grafts were histologically evaluated and expression of interleukins (IL)-17, -6 and -8, monocyte chemotactic protein (MCP)-1 and interferon (IFN)-gamma in serum and bronchoalveolar lavage (BAL) fluid, intracellular IL-4, -17, and IFN-gamma, in spleen lymphocytes were analysed. The levels of IL-17 in the serum and BAL fluid were significantly higher in MCMV infected mice versus not-infected mice (P = 0.0286 and P = 0.007, respectively) and the BAL levels of IL-17 peaked in 9 days (P = 0.001). The IL-17 level in the BAL was correlated with the grade of lung interstitial inflammation (r = 0.554, P = 0.0144). Serum IFN-gamma levels were also higher after infection than that in the not-infected mice (P = 0.0286). IL-17 production increases locally and systemically during MCMV interstitial pneumonia. Neutralization of IL-17 significantly suppressed lung inflammation at day14 as assessed by histology. These findings suggest that IL-17 is important in the pathology of MCMV interstitial pneumonia. PMID- 21623954 TI - Aberrant protein N-glycosylation impacts upon infection-related growth transitions of the haploid plant-pathogenic fungus Mycosphaerella graminicola. AB - The ascomycete fungus Mycosphaerella graminicola is the causal agent of Septoria Tritici Blotch disease of wheat and can grow as yeast-like cells or as hyphae depending on environmental conditions. Hyphal growth is however essential for successful leaf infection. A T-DNA mutagenesis screen performed on haploid spores identified a mutant, which can undergo yeast-like growth but cannot switch to hyphal growth. For this reason the mutant was non-pathogenic towards wheat leaves. The gene affected, MgAlg2, encoded a homologue of Saccharomyces cerevisiae ScAlg2, an alpha-1,2-mannosyltransferase, which functions in the early stages of asparagine-linked protein (N-) glycosylation. Targeted gene deletion and complementation experiments confirmed that loss of MgAlg2 function prevented the developmental growth switch. MgAlg2 was able to functionally complement the S. cerevisiae ScAlg2-1 temperature sensitive growth phenotype. Spores of DeltaMgAlg2 mutants were hypersensitive to the cell wall disrupting agent Calcofluor white and produced abnormally hypo-N-glycosylated proteins. Gene expression, proteome and glycoproteome analysis revealed that DeltaMgAlg2 mutant spores show responses typically associated with the accumulation of mis-folded proteins. The data presented highlight key roles for protein N-glycosylation in regulating the switch to hyphal growth, possibly as a consequence of maintaining correct folding and localization of key proteins involved in this process. PMID- 21623955 TI - Studies of locomotor network neuroprotection by the selective poly(ADP-ribose) polymerase-1 inhibitor PJ-34 against excitotoxic injury to the rat spinal cord in vitro. AB - Delayed neuronal destruction after acute spinal injury is attributed to excitotoxicity mediated by hyperactivation of poly(ADP-ribose) polymerase-1 (PARP 1) that induces 'parthanatos', namely a non-apoptotic cell death mechanism. With an in vitro model of excitotoxicity, we have previously observed parthanatos of rat spinal cord locomotor networks to be decreased by a broad spectrum PARP-1 inhibitor. The present study investigated whether the selective PARP-1 inhibitor N-(6-oxo-5,6-dihydrophenanthridin-2-yl)-(N,N-dimethylamino)acetamide.HCl (PJ-34) not only protected networks from kainate-evoked excitotoxicity, but also prevented loss of locomotor patterns recorded as fictive locomotion from lumbar (L) ventral roots (VRs) 24 h later. PJ-34 (60 MUm) blocked PARP-1 activation and preserved dorsal, central and ventral gray matter with maintained reflex activity even after a large dose of kainate. Fictive locomotion could not, however, be restored by either electrical stimulation or bath-applied neurochemicals (N methyl-D-aspartate plus 5-hydroxytryptamine). A low kainate concentration induced less histological damage that was widely prevented by PJ-34. Nonetheless, fictive locomotion was observed in just over 50% of preparations whose histological profile did not differ (except for the dorsal horn) from those lacking such a rhythm. Our data show that inhibition of PARP-1 could amply preserve spinal network histology after excitotoxicity, with return of locomotor patterns only when the excitotoxic stimulus was moderate. These results demonstrated divergence between histological and functional outcome, implying a narrow borderline between loss of fictive locomotion and neuronal preservation. Our data suggest that either damage of a few unidentified neurons or functional network inhibition was critical for ensuring locomotor cycles. PMID- 21623956 TI - Neutralization of interleukin-1beta reduces cerebral edema and tissue loss and improves late cognitive outcome following traumatic brain injury in mice. AB - Increasing evidence suggests that interleukin-1beta (IL-1beta) is a key mediator of the inflammatory response following traumatic brain injury (TBI). Recently, we showed that intracerebroventricular administration of an IL-1beta-neutralizing antibody was neuroprotective following TBI in mice. In the present study, an anti IL-1beta antibody or control antibody was administered intraperitoneally following controlled cortical injury (CCI) TBI or sham injury in 105 mice and we extended our histological, immunological and behavioral analysis. First, we demonstrated that the treatment antibody reached target brain regions of brain injured animals in high concentrations (> 11 nm) remaining up to 8 days post-TBI. At 48 h post-injury, the anti-IL-1beta treatment attenuated the TBI-induced hemispheric edema (P < 0.05) but not the memory deficits evaluated using the Morris water maze (MWM). Neutralization of IL-1beta did not influence the TBI induced increases (P < 0.05) in the gene expression of the Ccl3 and Ccr2 chemokines, IL-6 or Gfap. Up to 20 days post-injury, neutralization of IL-1beta was associated with improved visuospatial learning in the MWM, reduced loss of hemispheric tissue and attenuation of the microglial activation caused by TBI (P < 0.05). Motor function using the rotarod and cylinder tests was not affected by the anti-IL-1beta treatment. Our results suggest an important negative role for IL-1beta in TBI. The improved histological and behavioral outcome following anti IL-1beta treatment also implies that further exploration of IL-1beta-neutralizing compounds as a treatment option for TBI patients is warranted. PMID- 21623957 TI - Neurotrophin-4/5 is implicated in the enhancement of axon regeneration produced by treadmill training following peripheral nerve injury. AB - The role of neurotrophin-4/5 (NT-4/5) in the enhancement of axon regeneration in peripheral nerves produced by treadmill training was studied in mice. Common fibular nerves of animals of the H strain of thy-1-YFP mice, in which a subset of axons in peripheral nerves is marked by the presence of yellow fluorescent protein, were cut and surgically repaired using nerve grafts from non-fluorescent mice. Lengths of profiles of fluorescent regenerating axons were measured using optical sections made through whole mounts of harvested nerves. Measurements from mice that had undergone 1 h of daily treadmill training at modest speed (10 m/min) were compared with those of untrained (control) mice. Modest treadmill training resulted in fluorescent axon profiles that were nearly twice as long as controls at 1, 2 and 4 week survival times. Similar enhanced regeneration was found when cut nerves of wild type mice were repaired with grafts from NT-4/5 knockout mice or grafts made acellular by repeated freezing/thawing. No enhancement was produced by treadmill training in NT-4/5 knockout mice, irrespective of the nature of the graft used to repair the cut nerve. Much as had been observed previously for the effects of brief electrical stimulation, the effects of treadmill training on axon regeneration in cut peripheral nerves are independent of changes produced in the distal segment of the cut nerve and depend on the promotion of axon regeneration by changes in NT-4/5 expression by cells in the proximal nerve segment. PMID- 21623958 TI - Caprospinol: discovery of a steroid drug candidate to treat Alzheimer's disease based on 22R-hydroxycholesterol structure and properties. AB - The overall ability of the brain to synthesise neuroactive steroids led us to the identification of compounds that would reproduce aspects of neurosteroid pharmacology. The rate-determining step in neurosteroid biosynthesis is the import of the substrate cholesterol into the mitochondria, where it is metabolised into pregnenolone via the intermediate 22R-hydroxycholesterol. The levels of translocator protein 18-kDa, mediating the import of cholesterol into mitochondria, correlated with increased pregnenolone formation and reduced levels of 22R-hydroxycholesterol in biopsies from Alzheimer's disease (AD), but not age matched control, brains. 22R-hydroxycholesterol was shown to protect against beta amyloid (Abeta(42) )-induced neurotoxicity. In search of 22R-hydroxycholesterol stable analogues, we identified the naturally occurring heterospirostenol, (22R,25R)-20alpha-spirost-5-en-3beta-yl hexanoate (caprospinol) and derivatives that protect neuronal cells against Abeta(1-42) neurotoxicity. The neuroprotective effect of caprospinol is the result of a combination of overlapping properties, including: (i) the ability to bind to Abeta(42) and reduce plaque formation in the brain in vivo; (ii) interaction with components of the mitochondria respiratory chain resulting in an anti-uncoupling effect; (iii) the capacity to scavenge Abeta(42) monomers present in mitochondria; and (iv) the property of being a sigma-1 receptor ligand. In vivo, caprospinol crosses the blood-brain barrier, accumulates in the brain, and restores cognitive impairment in a pharmacological rat model of AD. Caprospinol is stable, does not bind to known steroid receptors, is devoid of mutagenic and genotoxic properties, and is devoid of acute toxicity in rodents. The pharmacokinetics and pharmacodynamics of caprospinol were studied, and long-term toxicity studies are under investigation, aiming to develop this compound as a disease-modifying drug for the treatment of AD. PMID- 21623959 TI - Evidence for RGS4 modulation of melatonin and thyrotrophin signalling pathways in the pars tuberalis. AB - In mammals, the pineal hormone melatonin is secreted nocturnally and acts in the pars tuberalis (PT) of the anterior pituitary to control seasonal neuroendocrine function. Melatonin signals through the type 1 Gi-protein coupled melatonin receptor (MT1), inhibiting adenylate cyclase (AC) activity and thereby reducing intracellular concentrations of the second messenger, cAMP. Because melatonin action ceases by the end of the night, this allows a daily rise in cAMP levels, which plays a key part in the photoperiodic response mechanism in the PT. In addition, melatonin receptor desensitisation and sensitisation of AC by melatonin itself appear to fine-tune this process. Opposing the actions of melatonin, thyroid-stimulating hormone (TSH), produced by PT cells, signals through its cognate Gs-protein coupled receptor (TSH-R), leading to increased cAMP production. This effect may contribute to increased TSH production by the PT during spring and summer, and is of considerable interest because TSH plays a pivotal role in seasonal neuroendocrine function. Because cAMP stands at the crossroads between melatonin and TSH signalling pathways, any protein modulating cAMP production has the potential to impact on photoperiodic readout. In the present study, we show that the regulator of G-protein signalling RGS4 is a melatonin-responsive gene, whose expression in the PT increases some 2.5-fold after melatonin treatment. Correspondingly, RGS4 expression is acutely sensitive to changing day length. In sheep acclimated to short days (SP, 8 h light/day), RGS4 expression increases sharply following dark onset, peaking in the middle of the night before declining to basal levels by dawn. Extending the day length to 16 h (LP) by an acute 8-h delay in lights off causes a corresponding delay in the evening rise of RGS4 expression, and the return to basal levels is delayed some 4 h into the next morning. To test the hypothesis that RGS4 expression modulates interactions between melatonin- and TSH-dependent cAMP signalling pathways, we used transient transfections of MT1, TSH-R and RGS4 in COS7 cells along with a cAMP-response element luciferase reporter (CRE-luc). RGS4 attenuated MT1-mediated inhibition of TSH-stimulated CRE-luc activation. We propose that RGS4 contributes to photoperiodic sensitivity in the morning induction of cAMP-dependent gene expression in the PT. PMID- 21623960 TI - Elevated corticosterone levels during the first postpartum period influence subsequent pregnancy outcomes and behaviours of the dam. AB - Postpartum depression affects 15% of new mothers and previous depressive episodes increase the risk for postpartum depression. Chronic administration of corticosterone (CORT) to dams during the postpartum period causes depressive-like behaviour and has been used as a model of postpartum depression. To better understand the subsequent progress of this model, we examined whether there were persistent effects of CORT treatment during the dam's first postpartum period on maternal care and mood following a subsequent pregnancy. Sprague-Dawley female rats received either sesame oil (control) or CORT (40 mg/kg) injections for 22 days during their first postpartum period. Subsequently, all females were re mated for a second time and neither group received treatment during the second postpartum period. Maternal care was observed from days 2-8 of each postpartum period and dams were tested in the forced-swim test on days 21 and 22 of the first and days 4 and 21 of the second postpartum period. As expected, the amount of time spent immobile in the forced-swim test was increased in CORT dams at the end of the first postpartum period; however, the amount of time spent immobile was decreased at the end of the second postpartum period relative to oil dams. Furthermore, dams treated with CORT in first postpartum period gave birth to a smaller litter with a larger male/female sex ratio after their second pregnancy. This implies that elevated stress hormone levels during the first postpartum period have a substantial influence on subsequent postpartum behaviour and litter characteristics. Further investigations are necessary to fully understand the effect of parity, experience during first motherhood, and hypothalamic-pituitary adrenal axis regulation on postpartum depression. PMID- 21623961 TI - Rapid effects of aggressive interactions on aromatase activity and oestradiol in discrete brain regions of wild male white-crowned sparrows. AB - Testosterone is critical for the activation of aggressive behaviours. In many vertebrate species, circulating testosterone levels rapidly increase after aggressive encounters during the early or mid-breeding season. During the late breeding season, circulating testosterone concentrations did not change in wild male white-crowned sparrows after an aggressive encounter and, in these animals, changes in local neural metabolism of testosterone might be more important than changes in systemic testosterone levels. Local neural aromatisation of testosterone into 17beta-oestradiol (E(2)) often mediates the actions of testosterone, and we hypothesised that, in the late breeding season, brain aromatase is rapidly modulated after aggressive interactions, leading to changes in local concentrations of E(2). In the present study, wild male white-crowned sparrows in the late breeding season were exposed to simulated territorial intrusion (STI) (song playback and live decoy) or control (CON) for 30 min. STI significantly increased aggressive behaviours. Using the Palkovits punch technique, 13 brain regions were collected. There was high aromatase activity in several nuclei, although enzymatic activity in the CON and STI groups did not differ in any region. E(2) concentrations were much higher in the brain than the plasma. STI did not affect circulating levels of E(2) but rapidly reduced E(2) concentrations in the hippocampus, ventromedial nucleus of the hypothalamus and bed nucleus of the stria terminalis. Unexpectedly, there were no correlations between aromatase activity and E(2) concentrations in the brain, nor were aromatase activity or brain E(2) correlated with aggressive behaviour or plasma hormone levels. This is one of the first studies to measure E(2) in microdissected brain regions, and the first study to do so in free-ranging animals. These data demonstrate that social interactions have rapid effects on local E(2) concentrations in specific brain regions. PMID- 21623962 TI - Osteoprotegerin levels predict mortality in patients with symptomatic aortic stenosis. AB - OBJECTIVES: To examine the prognostic value of osteoprotegerin (OPG) levels in relation to all-cause mortality in patients with symptomatic severe aortic stenosis (AS). DESIGN: We measured plasma OPG levels in 136 patients with symptomatic severe AS and investigated associations with transvalvular gradients, valve area, valve calcification (using ultrasonic backscatter analysis as an estimate) and measures of heart failure. Then, we assessed the prognostic value of elevated plasma OPG in determining all-cause mortality (n = 29) in these patients. RESULTS: Elevated OPG was poorly correlated with the degree of AS but was associated with increased backscatter measurements and impaired cardiac function. Furthermore, OPG was associated with all-cause mortality in patients with symptomatic AS, even after adjustment for conventional risk markers. The strongest association was obtained by using a combination of high levels of both OPG and N-terminal pro-brain natriuretic peptide (NT-proBNP), suggesting that these markers may reflect distinct pathways in the development and progression of AS. CONCLUSION: The level of circulating OPG is significantly associated with all cause mortality alone and in combination with NT-proBNP in patients with severe symptomatic AS. PMID- 21623963 TI - The effect of ezetimibe, administered alone or in combination with simvastatin, on lymphocyte cytokine release in patients with elevated cholesterol levels. AB - OBJECTIVE: Studies assessing the extra-lipid effects of ezetimibe have provided contrasting results. In the present study, we compared the effects of ezetimibe and simvastatin, administered alone or in combination, on the secretory function of human lymphocytes, systemic inflammation and endothelial function in subjects with elevated cholesterol levels. METHODS: A prospective study involving a group of 178 ambulatory patients with isolated hypercholesterolaemia who were randomly assigned in a double-blind fashion to 90days of treatment with ezetimibe (10mg), simvastatin (40mg), ezetimibe (10mg) plus simvastatin (4mg) or placebo. A total of 170 patients completed the study. MAIN OUTCOME MEASURES: Lymphocyte cytokine release and plasma levels of high-sensitivity C-reactive protein (hsCRP) and intercellular adhesion molecule 1 (ICAM-1). RESULTS: Although both drugs reduced lymphocyte release of tumour necrosis factor-alpha, interferon-gamma and interleukin-2 in a lipid-independent manner, only the effect of simvastatin was statistically significant (P<0.001). This lymphocyte-suppressing effect, which was accompanied by a decrease in plasma levels of hsCRP and ICAM-1 (P<0.001), was strongest in patients receiving both simvastatin and ezetimibe. There were no differences in lymphocyte-suppressing, systemic anti-inflammatory and endothelial protective effects of simvastatin between insulin-resistant and insulin-sensitive subjects, whereas the effects of ezetimibe and the combined treatment were greater in the former group of patients (P<0.01 and P<0.001, respectively). CONCLUSIONS: The results of this study indicate that simvastatin is superior to ezetimibe in producing lymphocyte-suppressing, systemic anti-inflammatory and endothelial protective effects in patients with elevated cholesterol levels. Hypercholesterolaemic patients with high cardiovascular risk may receive the greatest benefits from concomitant treatment with a statin and ezetimibe. PMID- 21623965 TI - A new approach to the isolation and characterization of wheat flour allergens. AB - BACKGROUND: The incidence of food allergy to wheat is increasing. Its diagnosis depends on the purity of major allergens and their inclusion in tests. Isolation and characterization of wheat allergens are therefore of utmost importance. OBJECTIVE: To purify and identify wheat flour allergens most frequently recognized by patients' IgE antibodies and to study their allergenicity. METHODS: Water/salt-soluble extracts from wheat flour were prepared and separated using a combination of ultrafiltration, isoelectric focusing and liquid chromatography. Purified proteins were analysed by immunoblotting using pooled sera from patients with atopic dermatitis who possessed IgE specific to wheat. Wheat proteins found to bind IgE were subsequently identified by matrix-assisted laser desorption/ionisation-time of flight mass spectrometry. The frequency and intensity of IgE binding of isolated proteins were tested using individual sera from patients and controls. RESULTS: We developed a procedure that allows isolation of wheat allergens from natural sources. Twenty-seven potential wheat allergens have been successfully identified; of these, the following seven are newly reported in food allergy: endogenous alpha-amylase/subtilisin inhibitor, trypsin/alpha-amylase inhibitor (AAI) CMX1/CMX3, thaumatin-like protein (TLP), xylanase inhibitor protein-1, beta-glucosidase, class II chitinase and 26 kDa endochitinase. TLP and wheatwin were shown to activate patients' basophils to a similar extent as two well-known allergens, lipid transfer protein (Tri a 14) and AAI 0.19 (Tri a 28.0101). CONCLUSION AND CLINICAL RELEVANCE: Our new approach enables the isolation of water/salt-soluble wheat allergens in their native form in amounts sufficient both for biological testing (in vivo and in vitro) and for physicochemical characterization. Such studies will lead to a more detailed knowledge of allergenicity of wheat proteins and to improved accuracy of diagnostic tests. PMID- 21623966 TI - Are TIM proteins involved in asthma development or pathology? PMID- 21623967 TI - Dietary polyphenols in the prevention and treatment of allergic diseases. AB - Allergic disorders encompass skin, food and respiratory allergies. Sensitization to a normally harmless allergen results in the immune system being biased to a predominant T-helper type 2 response. Re-exposure to the same allergen leads to a robust secretion of allergy-related mediators that eventually triggers symptoms. Our understanding of these disorders has enabled the search of therapeutic approaches that can either modulate the sensitization process or impact on allergic mediators, thus helping manage allergic symptoms. Polyphenols are one such class of compounds that are found in foods and plant sources and have been investigated for their anti-allergic effect in different disease models and in human clinical trials. Their anti-inflammatory profile is known to impact on the recruitment of immune cells to the skin and in preventing the development of secondary infections following disruption of the skin barrier. The interaction of polyphenols with proteins can modulate the process of allergic sensitization and their direct effect on allergic effector cells such as mast cells inhibit mediator release, resulting in the alleviation of symptoms. In addition, their endogenous anti-oxidant ability limits the extent of cellular injury from free radicals during the allergic insult. Overall, polyphenols hold promise as anti allergy agents capable of influencing multiple biological pathways and immune cell functions in the allergic immune response and deserve further investigation. The objective of the current review is to summarize the key findings and progress made in studying polyphenols as anti-allergic ingredients. Special emphasis is placed in this review to highlight key physiological, cellular and signalling pathways implicated in the mechanism of action of different polyphenols in the context of allergic disorders and their manifestations. PMID- 21623968 TI - Can exhaled inflammatory markers predict a steroid response in wheezing preschool children? AB - BACKGROUND: The efficacy of inhaled corticosteroids (ICS) varies among wheezing preschool children. Currently, it is not possible to predict which fraction of wheezing children will benefit from an ICS treatment. OBJECTIVE: We explored whether fractional exhaled nitric oxide (FeNO) and inflammatory markers in exhaled breath condensate (EBC) can predict an ICS response in preschool wheezers. METHODS: An 8-week ICS study (registered at Clinicaltrial.gov: NCT 00422747; 200 MUg; beclomethasone extra-fine daily) was performed in 93 wheezing children (age range 2.0-4.4 years). At baseline, FeNO was determined off-line. EBC was collected using a closed glass-condenser. The acidity of EBC was determined and other EBC markers [interleukin (IL)-1alpha, IL-2, IL-4, IL-5, IL 10, soluble intercellular adhesion molecule, interferon-gamma, eotaxin] were measured using a multiplex immunoassay. The change in airway resistance (Rint) and symptom score following ICS treatment was related to atopy (positive Phadiatop Infant test), FeNO and EBC markers. RESULTS: Airway resistance and symptoms mildly improved after ICS treatment [median (IQR): 1.4 (1.2-1.7) to 1.3 (1.1-1.5) kPa s/L, symptom score: 26 (23-28) to 28 (24-29), P < 0.01, respectively]. Only IL-10 and atopy had limited predictive value regarding a change in symptoms [beta (SE) =-0.13 (0.07), P = 0.08, beta (SE) = 2.05 (1.17), P = 0.08, respectively]. CONCLUSIONS AND CLINICAL RELEVANCE: We did not find convincing evidence that FeNO and EBC markers could predict an ICS response in preschool wheezers. Recommendations for future studies on this topic are given. PMID- 21623969 TI - Role of osteopontin, a multifunctional protein, in allergy and asthma. AB - Osteopontin (OPN) is an extracellular matrix protein and immune modulator with a wide range of functions. OPN is recognized as a key cytokine in Th1 immune responses, yet its potential involvement in allergic/asthmatic responses has been investigated only recently. Current data from molecular and cellular studies and studies of OPN-deficient mice provide evidence that OPN plays multiple roles in the regulation of allergic responses, including regulation of IgE response, inflammatory cell migration, and the development of airway fibrosis and angiogenesis. These results suggest that OPN is a pleiotropic cytokine that functions both systemically and locally in tissue mucosa. Notably, OPN is able to exert its effects through different functional domains, and the secreted and intracellular forms of OPN may have distinct functions. Future research to elucidate all aspects of OPN function is needed to ultimately establish its role in the regulation of immune responses and various disease processes, including those critically involved in the development of allergies and asthma. PMID- 21623970 TI - Air pollution and airway disease. AB - Epidemiological and toxicological research continues to support a link between urban air pollution and an increased incidence and/or severity of airway disease. Detrimental effects of ozone (O(3)), nitrogen dioxide (NO(2)) and particulate matter (PM), as well as traffic-related pollution as a whole, on respiratory symptoms and function are well documented. Not only do we have strong epidemiological evidence of a relationship between air pollution and exacerbation of asthma and respiratory morbidity and mortality in patients with chronic obstructive pulmonary disease (COPD), but recent studies, particularly in urban areas, have suggested a role for pollutants in the development of both asthma and COPD. Similarly, while prevalence and severity of atopic conditions appear to be more common in urban compared with rural communities, evidence is emerging that traffic-related pollutants may contribute to the development of allergy. Furthermore, numerous epidemiological and experimental studies suggest an association between exposure to NO(2) , O(3) , PM and combustion products of biomass fuels and an increased susceptibility to and morbidity from respiratory infection. Given the considerable contribution that traffic emissions make to urban air pollution researchers have sought to characterize the relative toxicity of traffic-related PM pollutants. Recent advances in mechanisms implicated in the association of air pollutants and airway disease include epigenetic alteration of genes by combustion-related pollutants and how polymorphisms in genes involved in antioxidant pathways and airway inflammation can modify responses to air pollution exposures. Other interesting epidemiological observations related to increased host susceptibility include a possible link between chronic PM exposure during childhood and vulnerability to COPD in adulthood, and that infants subjected to higher prenatal levels of air pollution may be at greater risk of developing respiratory conditions. While the characterization of pollutant components and sources promise to guide pollution control strategies, the identification of susceptible subpopulations will be necessary if targeted therapy/prevention of pollution-induced respiratory diseases is to be developed. PMID- 21623971 TI - Isolation of mesenchymal stem cells using the total length of umbilical cord for transplantation purposes. AB - BACKGROUND: Umbilical cord (UC) mesenchymal cells have the ability to differentiate into various cell types, which make them an easily obtainable source for therapeutic uses. Different approaches have been used to isolate mesenchymal stem cells (MSC). AIM: Here, we report a detailed enzymatic method where large number of cells can be efficiently isolated from the cord matrix and cryopreserved on the same day of arrival at the laboratory. METHODS/MATERIALS: Cells were successfully isolated from 12 samples, with a new procedure that uses the total length of the UC. MSC have been isolated using a detailed enzymatic method with collagenase and hyaluronidase followed by trypsin, without removing the vessels and without mincing the cord. Stem cells were measured with flow cytometry before cryopreservation and post-thaw. Cultured cells were assessed for MSC marker expression and adherence plasticity for three passages. Multilineage differentiation was performed. RESULTS: Nucleated cell yield was calculated at 0.95 * 10(6) /cm. MSC yield was calculated at 0.65 * 10(6) /cm of cord with flow cytometry while the mean length was 31 cm. Cultured cells expressed the mesenchymal markers CD29, CD90, CD105 and CD44. Mesenchymal marker expression remained intact over the three passages and post-thaw. Osteogenic and adipogenic differentiation was evaluated. CONCLUSIONS: Our findings provide a fast and efficient method for mesenchymal cell isolation from Wharton's jelly using the total length of the UC. This method resulted in a large number of cells while the cells retained their mesenchymal character after thawing. This method can be easily applied, along with UC blood, for UC banking. PMID- 21623972 TI - The key enzyme of sulfate assimilation, adenosine 5'-phosphosulfate reductase, is regulated by HY5 in Arabidopsis. AB - Plant sulfate assimilation is regulated by demand for reduced sulfur, as is its key enzyme, adenosine 5'-phosphosulfate reductase (APR). In a genetic screen for mutants lacking this regulation, we identified the bZIP transcription factor LONG HYPOCOTYL 5 (HY5) as a necessary component of the regulatory circuit. Regulation of APR activity by the inhibitor of glutathione synthesis, buthionine sulfoximine, or by the precursor of cysteine, O-acetylserine, was disrupted in the hy5 mutant. When dark-adapted plants were re-illuminated, the rapid induction of APR1 and APR2 mRNA levels was attenuated in hy5 seedlings, but APR3 regulation was not affected. Chromatin immunoprecipitation revealed that HY5 binds directly to the APR1 and APR2 promoters but not to the APR3 promoter. Accordingly, the regulation of APR1 and APR2 by O-acetylserine was disturbed in hy5 roots. HY5 is also important for the coordination of nitrogen and sulfur assimilation, as, unlike the wild-type, hy5 mutants do not undergo a reduction in sulfate uptake and APR activity during nitrogen starvation. Altogether, these data show that HY5 plays an important role in regulation of APR gene expression and plant sulfate assimilation. PMID- 21623973 TI - Dual functions of the nucleus-encoded factor TDA1 in trapping and translation activation of atpA transcripts in Chlamydomonas reinhardtii chloroplasts. AB - After endosymbiosis, organelles lost most of their initial genome. Moreover, expression of the few remaining genes became tightly controlled by the nucleus through trans-acting protein factors that are required for post-transcriptional expression (maturation/stability or translation) of a single (or a few) specific organelle target mRNA(s). Here, we characterize the nucleus-encoded TDA1 factor, which is specifically required for translation of the chloroplast atpA transcript that encodes subunit alpha of ATP synthase in Chlamydomonas reinhardtii. The sequence of TDA1 contains eight copies of a degenerate 38-residue motif, that we named octotrico peptide repeat (OPR), which has been previously described in a few other trans-acting factors targeted to the C. reinhardtii chloroplast. Interestingly, a proportion of the untranslated atpA transcripts are sequestered into high-density, non-polysomic, ribonucleoprotein complexes. Our results suggest that TDA1 has a dual function: (i) trapping a subset of untranslated atpA transcripts into non-polysomic complexes, and (ii) translational activation of these transcripts. We discuss these results in light of our previous observation that only a proportion of atpA transcripts are translated at any given time in the chloroplast of C. reinhardtii. PMID- 21623974 TI - The RCC1 family protein RUG3 is required for splicing of nad2 and complex I biogenesis in mitochondria of Arabidopsis thaliana. AB - We have identified a mitochondrial protein (RUG3) that is required for accumulation of mitochondrial respiratory chain complex I. RUG3 is related to human REGULATOR OF CHROMOSOME CONDENSATION 1 (RCC1) and Arabidopsis UV-B RESISTANCE 8 (UVR8). Although the family of RCC1-like proteins in Arabidopsis has over 20 members, UVR8 is the sole plant representative of this family to have been functionally characterized. Mitochondria from Arabidopsis plants lacking a functional RUG3 gene showed greatly reduced complex I abundance and activity. In contrast, accumulation of complexes III, IV and V of the oxidative phosphorylation system and the capacity for succinate-dependent respiration were unaffected. A comprehensive study of processes contributing to complex I biogenesis in rug3 mutants revealed that RUG3 is required for efficient splicing of the nad2 mRNA, which encodes a complex I subunit. A comparison of the formation of complex I assembly intermediates between rug3 and wild type mitochondria indicated that NAD2 enters the assembly pathway at an early stage. Remarkably, rug3 mutants displayed increased capacities for import of nucleus encoded mitochondrial proteins into the organelle and showed moderately increased mitochondrial transcript levels. This observation is consistent with global transcript changes indicating enhanced mitochondrial biogenesis in the rug3 mutant in response to the complex I defect. PMID- 21623975 TI - BON1 interacts with the protein kinases BIR1 and BAK1 in modulation of temperature-dependent plant growth and cell death in Arabidopsis. AB - The Arabidopsis copine gene BON1 encodes a calcium-dependent phospholipid-binding protein involved in plant growth homeostasis and disease resistance. However, the biochemical and molecular mechanisms by which BON1 modulates plant growth and defense responses are not well understood. Here, we show that BON1 interacts physically with the leucine-rich-repeat receptor-like kinases BIR1 (BAK1 interacting receptor-like kinase 1) and pathogen-associated molecular pattern (PAMP) receptor regulator BAK1 in vitro and in vivo. Additionally, bon1 and bir1 mutants exhibit synergistic interaction. While a bir1 null mutant has similar growth and cell-death defects compared with bon1, a bir1 bon1 double mutant displays more severe phenotypes than does the single mutants. The bon1-1 and bir1 1 phenotypes are partially suppressed by overexpression of BIR1 and BON1, respectively. Furthermore, the bir1 phenotype is attenuated by a loss-of-function mutation in the resistance (R) gene SNC1 (Suppressor of npr1-1, constitutive 1), which mediates defense responses in bon1. Intriguingly, BON1 and BIR1 can be phosphorylated by BAK1 in vitro. Our findings suggest that BIR1 functions as a negative regulator of plant resistance and that BON1 and BIR1 might modulate both PAMP- and R protein-triggered immune responses. PMID- 21623976 TI - Integrating long-day flowering signals: a LEAFY binding site is essential for proper photoperiodic activation of APETALA1. AB - The transition to flowering in Arabidopsis is characterized by the sharp and localized upregulation of APETALA1 (AP1) transcription in the newly formed floral primordia. Both the flower meristem-identity gene LEAFY (LFY) and the photoperiod pathway involving the FLOWERING LOCUS T (FT) and FD genes contribute to this upregulation. These pathways have been proposed to act independently but their respective contributions and mode of interaction have remained elusive. To address these questions, we studied the AP1 regulatory region. Combining in vitro and in vivo approaches, we identified which of the three putative LFY binding sites present in the AP1 promoter is essential for its activation by LFY. Interestingly, we found that this site is also important for the correct photoperiodic-dependent upregulation of AP1. In contrast, a previously proposed putative FD-binding site appears dispensable and unable to bind FD and we found no evidence for FD binding to other sites in the AP1 promoter, suggesting that the FT/FD-dependent activation of AP1 might be indirect. Altogether, our data give new insight into the interaction between the FT and LFY pathways in the upregulation of AP1 transcription under long-day conditions. PMID- 21623977 TI - Revealing impaired pathways in the an11 mutant by high-throughput characterization of Petunia axillaris and Petunia inflata transcriptomes. AB - Petunia is an excellent model system, especially for genetic, physiological and molecular studies. Thus far, however, genome-wide expression analysis has been applied rarely because of the lack of sequence information. We applied next generation sequencing to generate, through de novo read assembly, a large catalogue of transcripts for Petunia axillaris and Petunia inflata. On the basis of both transcriptomes, comprehensive microarray chips for gene expression analysis were established and used for the analysis of global- and organ-specific gene expression in Petunia axillaris and Petunia inflata and to explore the molecular basis of the seed coat defects in a Petunia hybrida mutant, anthocyanin 11 (an11), lacking a WD40-repeat (WDR) transcription regulator. Among the transcripts differentially expressed in an11 seeds compared with wild type, many expected targets of AN11 were found but also several interesting new candidates that might play a role in morphogenesis of the seed coat. Our results validate the combination of next-generation sequencing with microarray analyses strategies to identify the transcriptome of two petunia species without previous knowledge of their genome, and to develop comprehensive chips as useful tools for the analysis of gene expression in P. axillaris, P. inflata and P. hybrida. PMID- 21623978 TI - Comparison of oral-health-related quality of life during treatment with headgear and functional appliances. AB - BACKGROUND: Functional and headgear are two well-known approaches in the treatment of skeletal class II malocclusion in preadolescent children. Assessment of psycho-social impacts of wearing devices during the treatment period is central to enhancing the quality of healthcare services. AIM: This study aimed to compare oral-health-related quality of life in two groups consisting of children wearing headgear or functional appliances. We also compared these groups with a non-malocclusion group. DESIGN: The study population consisted of 187, 11- to 14 year-old children in three groups of functional (n = 67), headgear (n = 67) and non-malocclusion (n = 53). Children were asked to complete the translated version of the short form of the Child Perceptions Questionnaire. Total scores and subscale scores of the three clinical groups were compared through ANOVA. RESULTS: There was no significant difference in mean total scale score and subscale scores between functional and headgear groups (P > 0.05). Significant differences were found in both mean total and subscale scores between the malocclusion and non-malocclusion groups (P < 0.001) except oral symptoms subscale (P > 0.05). CONCLUSIONS: The results of this study reveal that functional and headgear appliances do not differ in terms of impact on daily life during the treatment. Moreover, both groups have poorer OHQoL compared to malocclusion group. PMID- 21623979 TI - Primary Sjogren syndrome in a 2-year-old patient: role of the dentist in diagnosis and dental management with a 6-year follow-up. AB - BACKGROUND. Primary Sjogren syndrome is a rare autoimmune disease, especially in children, mainly affecting girls (77%), and usually diagnosed around 10 years of age. Diagnosis during childhood is difficult, especially because of the diversity of the clinical presentation and difficulty obtaining reliable history data, accounting for a higher frequency of underdiagnosed cases. Differential conditions should be considered, especially the ones that promote xerostomia, such as diabetes, ectodermal dysplasia, rheumatoid arthritis, scleroderma, systemic lupus erythematosus, sarcoidosis, lymphoma, HIV and HTLV infection. Conditions associated with parotid enlargement should also be excluded, including juvenile recurrent parotitis (JRP), sialadenosis, sarcoidosis, lymphoma, infectious parotitis caused by streptococcal and staphylococcal infections, viral infections (paramyxovirus, Epstein-Barr virus, cytomegalovirus, and parvovirus), and diffuse infiltrative lymphocytosis syndrome (associated with HIV infection), and rare congenital conditions, such as polycystic parotid disease. CASE REPORT. A paediatric female patient was referred to our clinic for dental treatment complaining about dry mouth, oral discomfort, and dysphagia. The patient presented five of the required criteria to establish the diagnosis of pSS, including ocular symptoms, oral symptoms, evidence of keratoconjunctivitis sicca, focal sialadenitis confirmed by minor salivary gland biopsy, and evidence of major salivary gland involvement. Our patient did not have positive SS-A and SS-B autoantibodies. According to the literature, about 29% of individuals with pSS can present seronegativity for SS-A (anti-Ro) antibodies and about 33% can present seronegativity for SS-B (anti-La) antibodies. CONCLUSION. To the best of our knowledge, this is the youngest patient reported in the scientific English literature with pSS. Primary Sjogren syndrome has a wide clinical and immunologic spectrum and may progress with increased morbidity. Clinicians must be aware of the development of pSS in such an early age and exclude all possible differential findings to provide early diagnosis and treatment. PMID- 21623980 TI - Is sexual selection driving diversification of the bioluminescent ponyfishes (Teleostei: Leiognathidae)? AB - Sexual selection may facilitate genetic isolation among populations and result in increased rates of diversification. As a mechanism driving diversification, sexual selection has been invoked and upheld in numerous empirical studies across disparate taxa, including birds, plants and spiders. In this study, we investigate the potential impact of sexual selection on the tempo and mode of ponyfish evolution. Ponyfishes (Leiognathidae) are bioluminescent marine fishes that exhibit sexually dimorphic features of their unique light-organ system (LOS). Although sexual selection is widely considered to be the driving force behind ponyfish speciation, this hypothesis has never been formally tested. Given that some leiognathid species have a sexually dimorphic LOS, whereas others do not, this family provides an excellent system within which to study the potential role of sexual selection in diversification and morphological differentiation. In this study, we estimate the phylogenetic relationships and divergence times for Leiognathidae, investigate the tempo and mode of ponyfish diversification, and explore morphological shape disparity among leiognathid clades. We recover strong support for a monophyletic Leiognathidae and estimate that all major ponyfish lineages evolved during the Paleogene. Our studies of ponyfish diversification demonstrate that there is no conclusive evidence that sexually dimorphic clades are significantly more species rich than nonsexually dimorphic lineages and that evidence is lacking to support any significant diversification rate increases within ponyfishes. Further, we detected a lineage-through-time signal indicating that ponyfishes have continuously diversified through time, which is in contrast to many recent diversification studies that identify lineage-through-time patterns that support mechanisms of density-dependent speciation. Additionally, there is no evidence of sexual selection hindering morphological diversity, as sexually dimorphic taxa are shown to be more disparate in overall shape morphology than nonsexually dimorphic taxa. Our results suggest that if sexual selection is occurring in ponyfish evolution, it is likely acting only as a genetic isolating mechanism that has allowed ponyfishes to continuously diversify over time, with no overall impact on increases in diversification rate or morphological disparity. PMID- 21623981 TI - Temperature-related natural selection in a wind-pollinated tree across regional and continental scales. AB - Adaptive genetic variation is a key factor in evolutionary biology, but the detection of signatures of natural selection remains challenging in nonmodel organisms. We used a genome scan approach to detect signals of natural selection in the Black alder (Alnus glutinosa), a widespread wind-pollinated tree. Gene flow through pollen dispersal is believed to be high in this species, and we therefore expected to find a clear response to natural selection. In combination with two different landscape genetic approaches, we determined which environmental variables were most associated with the inferred selection. This analysis was performed on a regional scale (northern Belgium) and on a continental scale (Europe). Because climate-related differences are much more pronounced at the continental scale, we expected to find more selection-sensitive genetic markers across Europe than across northern Belgium. At both spatial scales, a substantial number of genetic loci were considered outliers, with respect to neutral expectations, and were therefore identified as selective. Based on results from our combined approach, four putative selective loci (or 2.5%) were recovered with high statistical support. Although these loci seemed to be associated with different environmental variables, they were mainly temperature-related. Our study demonstrates that the use of complementary methods in landscape genetics allows the discovery of selective loci which otherwise might stay hidden. In combination with a genome scan, the selective loci can be verified and the nature of the selection pressure can be identified. PMID- 21623982 TI - Cognitive, psychomotor and polysomnographic effects of trazodone in primary insomniacs. AB - Trazodone is prescribed widely as a sleep aid, although it is indicated for depression, not insomnia. Its daytime cognitive and psychomotor effects have not been investigated systematically in insomniacs. The primary goal of this study was to quantify, in primary insomniacs, the hypnotic efficacy of trazodone and subsequent daytime impairments. Sixteen primary insomniacs (mean age 44 years) participated, with insomnia confirmed by overnight polysomnography (sleep efficiency <= 85%). Trazodone 50 mg was administered to participants 30 min before bedtime for 7 days in a 3-week, within-subjects, randomized, double-blind, placebo-controlled design. Subjective effects, equilibrium (anterior/posterior body sway), short-term memory, verbal learning, simulated driving and muscle endurance were assessed the morning after days 1 and 7 of drug administration. Sleep was evaluated with overnight polysomnography and modified Multiple Sleep Latency Tests (MSLT) on days 1 and 7. Trazodone produced small but significant impairments of short-term memory, verbal learning, equilibrium and arm muscle endurance across time-points. Relative to placebo across test days, trazodone was associated with fewer night-time awakenings, minutes of Stage 1 sleep and self reports of difficulty sleeping. On day 7 only, slow wave sleep was greater and objective measures of daytime sleepiness lower with trazodone than with placebo. Although trazodone is efficacious for sleep maintenance difficulties, its associated cognitive and motor impairments may provide a modest caveat to health care providers. PMID- 21623983 TI - Is it time to start discussing what type of computer-delivered intervention? PMID- 21623984 TI - Community capacity building in practice: constructing its meaning and relevance to health promoters. AB - Community capacity building (CCB) is held up as a benchmark for sustainable health promotion, reflecting the empowering discourse of the Ottawa Charter (WHO 1986). In light of concerns that this language may be that of the presiding bureaucratic elite rather than the realities of those working directly with communities (Laverack & Labonte 2000), we question whether CCB reflects the work of New Zealand health promoters. The aim of this study is to assess what CCB means to health promoters and how relevant it is to their work in New Zealand. Focus groups and interviews were carried out with 64 health promoters between January 2008 and March 2009. The results of this qualitative study indicated that, while the terminology of CCB is poorly established in New Zealand, the overwhelming majority of participants felt that, to be an effective health promoter, they needed the buy-in and support of the communities in which they work. As a result, community-driven approaches have emerged as a core component of good health promotion practice in New Zealand. Yet, the concept of CCB was applied loosely with health promoters adopting language and practices corresponding more with the nuances of community development. The limited use of systematic approaches to building community capacity was accompanied by few successes achieving sustainable health promotion programmes. In prioritising community relationships many health promoters were placed in an ideological bind whereby achieving community ownership over health promotion meant compromising the evidence base of their programmes. Academic discussions of CCB appear to have gained little traction into the realm of health promotion practice in New Zealand highlighting the need for relevant research with a strong grounding in practice. PMID- 21623985 TI - The psychosocial experience of parents receiving care closer to home for their ill child. AB - Current health policy in England regarding the management of childhood illness advocates for care to be delivered as close to home as possible. The aim of this article is to report findings from a qualitative component of a larger study evaluating models of care closer to home (CCTH) for children and young people who are ill. The focus is on parents' psychosocial experience of receiving CCTH for their ill child. A qualitative case study design was used, with four Primary Care Trusts in England as the case study sites. In-depth, semi-structured interviews were conducted with 27 parents and one extended family caregiver of children using CCTH services within the case study sites. Interviews were conducted face to-face and audio-recorded with permission. Data were collected in 2009. There was an overall preference for CCTH over hospital-based care where possible. The parents in this study experienced differing levels of responsibility as part of this care, and responded to this in different ways. Being supported emotionally and socially by practitioners was an important part of receiving CCTH for parents, especially when they had increased responsibility as part of this care. Developing relationships with practitioners appeared to be a medium through which parents received support. If the provision of CCTH continues to expand in line with current policy, provision of support for parents should be considered a fundamental aspect of service development. PMID- 21623986 TI - Workload in community pharmacies in the UK and its impact on patient safety and pharmacists' well-being: a review of the evidence. AB - New contractual frameworks for community pharmacy are believed to have increased workload for pharmacists; too much work has been implicated in high profile cases of dispensing errors leading to patient harm, and concerns about pharmacists' well-being. A review was undertaken to ascertain whether community pharmacists' workload has increased and whether links between workload and patient safety and pharmacists' well-being have been established. We searched Scopus; EMBASE; MEDLINE; PubMed; CINAHL; PsychINFO; ASSIA; E-pic, and International Pharmaceutical Abstracts for research published between 1989 and 2010 containing data on UK community pharmacy workload, and on its consequences when workload was found to be a determinant of either patient or pharmacist outcomes. Researchers assessed retrieved material against inclusion and exclusion criteria and synthesised findings using a data extraction form. Fifteen studies were retrieved that met the inclusion criteria. A number of methodological weaknesses were identified: studies categorised work tasks and workload differently making comparisons over time or between studies difficult; most studies were small scale or conducted in specific localities, or lacked sufficient methodological information to rule out bias; studies that control for possible confounders are rare. The reviewed research suggests that community pharmacists still spend the majority of their time involved in activities associated with the dispensing of prescriptions. There is some evidence that community pharmacists' workload has increased since the introduction of the new contracts in England and Wales, especially around the core activity of dispensing prescriptions and medicines use reviews. There is also some evidence to suggest a link between heavy workload and aspects of pharmacists' well-being but there is no robust evidence indicating threats to patient safety caused by their having too much work to do. More high quality research is required to examine what constitutes too much work, the impact of high workload, and associations with other work place factors. PMID- 21623987 TI - A unique preliminary study on placental apoptosis in mice with passive immunization of anti-phosphatidylethanolamine antibodies and anti-factor XII antibodies. AB - PROBLEM: Antiphospholipid antibodies have been investigated both in humans and in animal models. In contrast, there are fewer reports describing anti phosphatidylethanolamine (aPE) antibodies in humans, and there are no reports of animal studies with aPE till date. Clinically, FXII deficiency or anti-FXII antibodies are sometimes associated with aPE in patients with recurrent pregnancy loss. Therefore, we asked whether aPE and/or anti-FXII in mice could cause fetal resorption, placental thrombosis and apoptosis. Moreover, antibodies to respective target antigens (LDC27 or IPP30) could cause pregnancy failure as well. METHODS OF STUDY: Animal models were used to carry out these objectives. All the animals were immunized with different antibodies by passive immunization. Placental samples were used for various observations. RESULTS AND CONCLUSIONS: Mice with passive immunization of aPE (or anti-LDC27) and aFXII (or anti-IPP30) produced a slight increase in fetal resorption, but markedly induced thrombosis and hemorrhage in the placenta associated with lower platelet counts and increased placental apoptosis. In addition, fewer mitotic cells, less trophoblast giant cell invasion, and more shrunken cells in the deciduas were seen. Our study supports the pathogenic role of aPE and aFXII in pregnancy complications and also suggests a beneficial role of LDC27 and IPP30 antigens on pregnancy failures. PMID- 21623988 TI - Transforming growth factor-beta1 gene polymorphisms in Korean women with endometriosis. AB - PROBLEM: To investigate the association between endometriosis, transforming growth factor-beta1 (TGFB1) gene polymorphisms, and serum TGF-beta1 levels in Korean women. METHOD OF STUDY: The -509C/T, 868T/C, 913G/C and 979G/A polymorphisms of the TGFB1 gene were analyzed in women with (n = 131) and without (n = 107) endometriosis using restriction fragment length polymorphism (RFLP) analysis. Serum TGF-beta1 levels were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: The 913G/C and 979G/A polymorphisms were not observed in the study participants. The genotype and allele distribution of the -509C/T and 868T/C polymorphisms in endometriosis were similar to those in controls. However, the -509T/868C (TC) haplotype allele was observed 4.55 times more frequently in early-stage endometriosis than in other haplotype alleles. Serum TGF-beta1 levels were significantly higher in endometriosis than in controls. The single and haplotype genotype of -509C/T and 868T/C polymorphisms were not related with serum TGF-beta1 levels. CONCLUSION: The TC haplotype allele of TGFB1-509C/T and 868T/C polymorphisms may be associated with early-stage endometriosis in Korean women. PMID- 21623989 TI - FTY720-induced conversion of conventional Foxp3- CD4+ T cells to Foxp3+ regulatory T cells in NOD mice. AB - PROBLEM: FTY720 is known as an agonist of sphingosine-1-phosphate (S1P) receptor, but little is known about the possibility that FTY720 induces the conversion of conventional Foxp3(-) CD4(+) T cells to Foxp3(+) regulatory T cells in non-obese diabetic (NOD) mice. METHOD OF STUDY: FTY720 treatment was performed using Foxp3( ) CD4(+) T cells purified from NOD mice. RESULTS: FTY720 caused an increase in Foxp3(+) Treg cells in lymphoid organs in NOD mice. FTY720 effectively induced Foxp3 expression in Foxp3(-) CD4(+) T cells both in vitro and in vivo, an effect that was inhibited by a TGF-beta-neutralizing antibody or the proinflammatory cytokine IL-6. T-cell-mediated embryo rejection in NOD mice was prevented upon FTY720 treatment. CONCLUSIONS: The use of FTY720 along with Ag administration may represent a useful therapeutic strategy to selectively expand Ag-specific Foxp3(+) Tregs to intervene autoimmune and infectious diseases. PMID- 21623990 TI - First evidence of genetic association between the MIF-173G/C single-nucleotide polymorphisms and polycystic ovary syndrome. AB - PROBLEM: The purpose of this study was to investigate whether polymorphism of MIF gene is associated with PCOS. METHOD OF STUDY: The MIF-173G/C single-nucleotide polymorphism (SNP) was detected in 529 PCOS patients and 585 healthy female controls of Chinese Han ancestry. The association of the gene variants with clinical and metabolic parameters and hormone levels was investigated. RESULTS: The frequencies of genotypes and allelotypes of the MIF-173G/C SNP did significantly differ between women with PCOS and healthy controls (P = 0.017 and P = 0.003, respectively). They did significantly differ between obese PCOS patients and obese controls (P = 0.029 and P = 0.039, respectively). The MIF-173 CC and CG genotypes were associated with higher body mass index (BMI) and waist to-hip ratio (WHR) in both PCOS patients (P < 0.001, P = 0.001) and normal controls (P < 0.001, P = 0.002). The PCOS patients with CC and CG genotypes had higher fasting plasma glucose levels (P < 0.001), higher fasting insulin levels (P < 0.001), and higher HOMA-IR (P < 0.001) compared with patients with the GG genotype. CONCLUSION: The MIF-173G/C polymorphism is associated with PCOS in Chinese Han women and may contribute to the phenotypic expression of PCOS. PMID- 21623991 TI - First trimester pregnancy decidual natural killer cells contain and spontaneously release high quantities of granulysin. AB - PROBLEM: Granulysin (GNLY) is a novel cytolytic protein lytic against a variety of tumor cells and microbes. The role of GNLY during pregnancy has not been extensively explored. The aim of this study is to examine GNLY expression and distribution in the first trimester pregnancy peripheral blood (PB) and decidua, the ability of decidual and PB natural killer (NK) cells to secrete GNLY spontaneously, and the role of antigen-presenting cells (APC) in the regulation of GNLY expression in decidual NK cells. METHOD OF STUDY: GNLY expression was analyzed using cell permeabilization method, flow cytometry, and immunohistochemistry. GNLY secretion by purified NK cells was detected by ELISA method. RESULTS: GNLY is abundantly expressed at the maternal-fetal interface in the first trimester pregnancy. Decidual T lymphocytes express significantly higher levels of GNLY (58%) then PB T lymphocytes (11%). Over 85% of decidual CD56(+) cells express GNLY and when cultured spontaneously release high quantities of GNLY. Decidual APC participate in the control of GNLY expression in CD56(+) cells. CONCLUSION: Abundant expression of GNLY in the decidual immunocompetent cells and the capacity of decidual CD56(+) cells to spontaneously secrete high quantities of GNLY point to important protective and immunomodulatory role that this molecule could play at the maternal-fetal interface. PMID- 21623992 TI - Increased reactive oxygen species and tumor necrosis factor-alpha production by monocytes are associated with elevated levels of uric acid in pre-eclamptic women. AB - PROBLEM: To evaluate associations between hyperuricemia and increases in production of reactive oxygen species (ROS) and tumor necrosis factor alpha (TNF alpha) in pre-eclamptic pregnancies. METHOD OF STUDY: This study investigated serum uric acid levels, monocyte production of TNF-alpha, superoxide anion (O(2)( )) and hydrogen peroxide (H(2)O(2)), as well as superoxide dismutase (SOD) and catalase (CAT) activities in erythrocytes from 30 women with pre-eclampsia (PE) compared with 30 normotensive (NT) pregnant women in the last trimester of pregnancy. RESULTS: Serum uric acid levels (6.1 versus 2.8 mg/dL) as well as endogenous O(2)(-) (2.2 versus 1.6 nm), H(2)O(2) (1.8 versus 1.4 nm) and TNF alpha (91.6 versus 40.4 pg/mL) released from monocytes were significantly higher in the pre-eclamptic group than in the NT group (P < 0.05). SOD activity in erythrocytes was also significantly elevated in the PE group (5969.2 versus 4834.7 U/g Hb). No significant difference between groups was observed in relation to CAT activity. CONCLUSIONS: Elevated serum uric acid levels are correlated with higher O(2)(-) and TNF-alpha production by monocytes in women with PE. This may contribute to the enhanced oxidative and inflammatory state characteristic of this disorder. PMID- 21623993 TI - Relevance of syndecan-1 in the trophoblastic BeWo cell syncytialization. AB - PROBLEM: To investigate the role of syndecan-1 in the differentiation of the BeWo cells into syncytiotrophoblast. METHOD OF STUDY: BeWo cells were stimulated with forskolin to form syncytia, and the expression of syndecan-1, desmoplakin I+II, human chorionic gonadotrophin (hCG) and angiogenesis-associated factors was analyzed. Syndecan-1 was silenced by siRNA to evaluate its involvement in the forskolin-mediated syncytia formation. RESULTS: Treatment of the BeWo cells with forskolin led to a significant increase in the syncytia formation. It was associated with an increase in the expression of syndecan-1 with a concomitant decrease in the expression of desmoplakin I+II. Forskolin treatment of the BeWo cells also led to an increase in the secretion of soluble endoglin, whereas no change was observed in the soluble fms-like tyrosine kinase-1. Silencing of the syndecan-1 expression in BeWo cells led to a significant decrease in cell fusion both in the presence and in the absence of forskolin. It was associated with a significant decrease in hCG level in the conditioned medium. CONCLUSION: Syndecan 1 is up-regulated in BeWo cells during differentiation and its silencing inhibits syncytialization and thus could be a useful biomarker for syncytiotrophoblast formation. PMID- 21623994 TI - Elevated preconception CD56+ 16+ and/or Th1:Th2 levels predict benefit from IVIG therapy in subfertile women undergoing IVF. AB - PROBLEM: We sought to answer two questions: First, is there a group of patients who benefit from intravenous immunoglobulin (IVIG) in IVF? Second can this group of patients be identified by preconception blood testing? METHOD OF STUDY: A total of 202 IVF cycles in subfertile women were divided into four groups. Group I: 62 cycles with preconception Th1:Th2 ratio and/or % CD56(+) cell elevation using IVIG; Group II: 27 cycles with similar Th1:Th2 and/or % CD56(+) cell elevation not using IVIG; Group III: 71 cycles with normal Th1:Th2 and/or % CD56(+) cell levels using IVIG; Group IV: 42 cycles with normal Th1:Th2 and % CD56(+) levels not using IVIG. These groups were similar with regard to patient age, diagnosis, and past failure history. RESULTS: The implantation rate (number of gestational sacs per embryo transferred, with an average of two embryos transferred per cycle) was 45% (55/123), 22% (12/54), 54% (75/139), and 48% (40/84) for Groups I-IV, respectively. The clinical pregnancy rate (fetal heart activity per IVF cycle started) was 61% (38/62), 26% (7/27), 69% (49/71), and 71% (30/42), respectively. The live birth rate was 58% (36/62), 22% (6/27), 61% (43/71), and 71% (30/42), respectively, and the live birth per embryo transferred was 40% (49/123), 13% (7/24), 43% (60/139), and 48% (40/84), respectively. There was a significant improvement in implantation, clinical pregnancy, live birth rate and live birth rate per embryo transferred for Group I versus Group II (P = 0.0032, 0.0021, 0.0017, and 0.0002, respectively) and for Group II versus Group IV (P = 0.0021, 0.0002, <0.0001 and <0.0001, respectively). There was no significant difference in success rates between Groups I and III (P = 0.085, 0.23, 0.45, 0.34, respectively) and between Groups III and IV (P = 0.22, 0.48, 0.17, 0.31, respectively). CONCLUSION: In subfertile women with preconception Th1:Th2 and/or % CD56(+) cell elevation, IVF success rates are low without IVIG therapy but significantly improve with IVIG therapy. In patients with normal Th1:Th2 and normal CD56(+) cell levels, IVF success rates were not further improved with IVIG therapy. IVIG may be a useful treatment option for patients with previous IVF failure and preconception Th1:Th2 and/or NK elevation. Preconception immune testing may be a critical tool for determining which patients will benefit from IVIG therapy. Prospective controlled studies (preferably double-blind, stratified, and randomized) are needed for confirmation. PMID- 21623995 TI - Elevated serum levels of interleukin-6, interleukin-1beta and human chorionic gonadotropin in pre-eclampsia. AB - PROBLEM: Pre-eclampsia (PE) is a pregnancy-specific syndrome of unknown aetiology. It is believed to involve an inflammatory process. The aim of the study was to investigate and compare the concentrations of two proinflammatory cytokines interleukin-6 (IL-6) and interleukin-1beta (IL-1beta) and to evaluate the possible interaction between them and human chorionic gonadotropin (hCG) in women with normotensive pregnancy and PE. METHOD OF STUDY: A prospective case control study was carried out in 30 women with PE and 30 normotensive controls. Serum IL-1beta, IL-6 and hCG levels were determined by enzyme-linked immunosorbent assay (ELISA) and automated immunofluorescent assay, respectively. RESULTS: Serum IL-6, IL-1beta and hCG levels were significantly increased in women with PE compared to controls (P < 0.001 for each); however, no correlation was found between IL-6, IL-1beta and hCG. CONCLUSION: Our results highlight the inflammatory origin of PE and reinforce the possible role of hCG in the complex aetiology of its pathogenesis. PMID- 21623996 TI - Celiac disease and gyneco-obstetrics complications: can serum antibodies modulate tissue transglutaminase functions and contribute to clinical pattern? AB - PROBLEM: Untreated celiac disease (CD) is often associated with early miscarriages, infertility, and alterations in menstrual cycle. Tissue transglutaminase (tTG) antibodies could be involved by interfering with tTG transamidating activity and/or biological functions mediated by its interaction with fibronectin (FN). METHOD OF STUDY: The correlation between the presence of extra-digestive disorders and the reactivity of sera against tTG-FN and its effects on tTG transamidating activity was analyzed in a group or 50 women with recently diagnosed CD. RESULTS: Heterogeneous behavior was observed among serum samples derived from patients with different complaints, suggesting that differences in fine specificity patterns could condition clinical outcome. Sera from women with gynecological and/or obstetric problems induced significant inhibition of in vitro enzymatic activity in comparison with those without these kinds of disorders. CONCLUSIONS: The significant correlation observed between serum effects and clinical profile suggests a putative involvement of tTG specific antibodies in gynecological and/or obstetric disorders during active CD. PMID- 21623997 TI - Thyroid autoimmunity and recurrent miscarriage. AB - PROBLEM: To investigate the role of antithyroid autoantibodies (ATA) in recurrent miscarriage (RM). METHODS: In this case-control study, a total of 160 women with RM and 100 healthy women were investigated for the presence of serum ATA directed against thyreoglobulin (TG-Ab), thyroid peroxidase (TPO-Ab) and TSH receptor (TSHr-Ab), which were determined by either chemiluminescence or radioimmunoassay. RESULTS: Antithyroid autoantibodies were detected in 46 (28.75%) women with RM and in 13 (13%) women of the control group (P < 0.05). The frequencies for TG-Ab and TPO-Ab were higher in RM than in control women. Among the women of RM group, 91.3% of ATA+ women were positive also for other autoantibodies. The majority of study women were euthyroid. CONCLUSIONS: Antithyroid autoantibodies, particularly TG-Ab, are associated with RM and could be an expression of a more general maternal immune system abnormality leading to RM. ATA could have a role in RM irrespective of thyroid hormone status. PMID- 21623998 TI - Urotensin-II 143 G/A polymorphism is not associated with the risk of preeclampsia in Korean women. AB - PROBLEM: Urotensin-II (UTS-II) may be associated with preeclampsia. We therefore assessed whether the UTS-II 143 G/A polymorphism is associated with preeclampsia in Korean women. METHOD OF STUDY: This prospective case-control study enrolled 109 patients with preeclampsia and 144 healthy pregnant controls, all of whom were genotyped for the UTS-II 143 G/A polymorphism using the real-time TaqMan SNP Genotyping Assay. RESULTS: There was no significant difference in the distribution of the 143 G/A polymorphism or the frequency of the 143 A allele between women with preeclampsia and controls. In a dominant model, carriers of the A allele were not significantly more prevalent in the preeclamptic (53.2%) than in the control (60.4%) group. In addition, subgroup analysis showed no significant difference in genotype distribution or allelic frequency of the 143 G/A polymorphism between women with mild or severe preeclampsia and controls. CONCLUSION: In Korean women, the common UTS-II 143 G/A polymorphism is unlikely to have an association with the risk of preeclampsia. PMID- 21623999 TI - Cluster analysis of placental inflammatory proteins can distinguish preeclampsia from preterm labor and premature membrane rupture in singleton deliveries less than 28 weeks of gestation. AB - PROBLEM: Inflammation within the preterm placenta is common and leads to adverse outcomes for premature infants. The risks of complications are different between iatrogenic (e.g. PE) and spontaneous (e.g. PL and membrane rupture) causes of preterm delivery, suggesting different underlying biology contributes to these placental conditions. METHOD OF STUDY: Thirty preterm singleton placentas from the following groups were analyzed: (i) severe PE, (ii) preterm premature membrane rupture (pPROM), and (iii) PL. Proinflammatory and anti-inflammatory cytokines, adhesion and angiogenic molecules were measured in placental lysates using a multiplex assay. K-means cluster analysis was used to generate patterns of protein level intensity. RESULTS: Three cluster patterns were apparent. Placentas from PE had high levels of vascular endothelial growth factor (VEGF) combined with low levels of acute inflammatory proteins (IL-1beta, IL-18, IL-6, TNF-alpha), low IL-1 RA, and high transforming growth factor beta (TGF-beta). PL and pPROM had higher anti-inflammatory IL-1 RA and thrombomodulin combined with lower VEGF, regardless of proinflammatory cytokines and adhesion molecules. Half of the PL and pPROM cases had clusters of heightened inflammatory responses (lower TGF-beta clustered with higher intensity of inflammatory mediators). CONCLUSION: Discriminating protein patterns were elucidated and may serve as a foundation from which to understand the biologic mechanisms underlying these pregnancy complications. PMID- 21624000 TI - Analysis of Treg cell population alterations in the peripheral blood of patients treated surgically for ovarian cancer - a preliminary report. AB - PROBLEM: Treg cells constitute the main cell population that enables cancer cells to evade immune surveillance. An alteration in the Treg cell population might correspond to the diminishment of the tumour mass in patients with cancer and could therefore be a useful marker of the intensity of the selective suppression of the host immune system and also of the degree of radicalism of a procedure. Certainly, it is well known that in order for anti-cancer therapy to succeed the proper immune response against cancer cells must be restored. Furthermore, monitoring the level of selective immune system suppression during cancer therapy might yield information that would support a decision to supplement standard therapy by immunotherapy or to increase the degree of radicalism of the applied therapy. METHOD OF STUDY: We examined the Treg cell populations in the peripheral blood of a group of patients treated surgically for ovarian cancer. In each patient, the peripheral blood samples were collected both prior to and 1 day after the surgical procedure, and then again 5 days after the procedure. The presence of regulatory T cells in the samples was analyzed by means of flow cytometry. RESULTS: In our study, the percentages of FOXP3(+) cells in the subpopulation of CD4(+) T lymphocytes found in the peripheral blood of the patients before the surgical intervention were statistically significantly higher than those observed in the peripheral blood of these same patients after the surgical procedure. CONCLUSION: It would seem that the alteration in the Treg cell subpopulation could be a key factor in determining the status of the tumour microenvironment. Most likely, it could provide information about whether the proper level of anti-cancer immune response could be restored. The possibility of restoring the immune response may directly correspond to the degree of radicalism of the surgical intervention. PMID- 21624001 TI - Effect of suspending viscosity on red blood cell dynamics and blood flows in microvessels. AB - To obtain a better understanding of the beneficial effect of high plasma viscosity observed in hemodilution and resuscitation experiments, we conducted a computational study to investigate the suspending viscosity effect on red blood cell (RBC) dynamics and blood flow behaviors in microvessels. For single RBCs in simple shear or channel flows, RBCs appear more flexible as indicated by the tank treading motion in shear flows and the strong transverse migration in channel flows. For the multiple RBC flows in straight channels, our results indicate no significant change with the suspending viscosity in stable flow structure and hemorheologic behaviors, under both constant flow and forcing conditions. However, due to the increase in apparent cell deformability in a more viscous medium, the cell-free layer (CFL) can be established in a shorter distance along the channel. Considering the multilevel bifurcated structure of the microvascular network, this change in CFL development distance may affect the phase skimming and RBC separation processes at the downstream bifurcation, and therefore the microcirculation performance in the tissue. This may suggest a possible mechanism for the high functional capillary density associated with a high suspending viscosity observed in experiments. PMID- 21624002 TI - Bronchial brushings for investigating airway inflammation and remodelling. AB - Asthma is the commonest medical cause for hospital admission for children in Australia, affects more than 300 million people worldwide, and is incurable, severe in large number and refractory to treatment in many. However, there have been no new significant treatments despite intense research and billions of dollars. The advancement in our understanding in this disease has been limited due to its heterogeneity, genetic complexity and has severely been hampered particularly in children by the difficulty in obtaining relevant target organ tissue. This review attempts to provide an overview of the currently used and recently developed/adapted techniques used to obtain lung tissue with specific reference to the airway epithelium. PMID- 21624003 TI - Spirometry values in a Greek population: is there an appropriate reference equation? AB - BACKGROUND AND OBJECTIVE: Most published reference values for lung function test (LFT) parameters introduce systematic bias. The aim of this study was to compare measured values of FEV(1) and FVC with the corresponding normal predicted values in a Greek population, and to produce reference equations for LFT parameters in this population. METHODS: In a cross-sectional study conducted in Macedonia, Greece, 1080 adult healthy, non-smokers (432 men, 648 women, aged 18-80 years), underwent spirometry. Measured values of FVC and FEV(1) were compared with predicted values determined using three existing sets of reference equations: one recently derived from a European population and two others widely used in Europe (European Coal and Steel Community; ECSC) and the USA (National Health and Nutrition Examination Survey; NHANES III). Height and age were entered into the multivariate regression analysis to produce reference equations for LFT parameters. RESULTS: All three published sets of equations underpredicted FEV(1) in men. FVC was accurately predicted by all equations except NHANES III. The discrepancy was even greater among women; the ECSC equation underpredicted both FEV(1) and FVC, the NHANES III equation overpredicted both FEV(1) and FVC, while the third set of equations accurately predicted FEV(1) but overpredicted FVC. The derived reference equation for FEV(1) in men was -0.28 * age + 0.057 * height - 4.91, and in women -0.021 * age + 0.039 * height - 2.58. The derived reference equation for FVC in men was -0.28 * age + 0.071 * height - 6.763, and in women 0.019 * age + 0.056 * height - 5.018. CONCLUSIONS: Measured FEV(1) and FVC values in a Greek population differed significantly from those predicted using previously published reference equations. The new locally derived spirometry reference equations may be more suitable for evaluation of lung function in everyday practice. PMID- 21624004 TI - Pulse steroids as induction therapy for children with ulcerative colitis. AB - BACKGROUND: Corticosteroids therapy, classically the first-line treatment for ulcerative colitis (UC), often causes serious side-effects. Theoretically, pulse steroid therapy where high doses are given for a shorter period may have maximal beneficial effects and minimal side-effects as induction therapy for UC. We have therefore retrospectively compared induction therapy using pulse steroids with conventional steroid treatment for children and adolescents with moderate-to severe UC. METHODS: We utilized conventional steroid treatment (prednisolone 1 1.5 mg/kg/day) as an induction treatment in 17 UC patients between 1985 and 2006. Alternatively we used a 3-day megadose pulse steroid therapy (methylprednisolone intravenously 20-30 mg/kg/day, max. 1000 mg/day) in 20 UC patients from 1993 to 2006. RESULTS: Pulse steroid therapy successfully induced rapid remission in UC patients with moderate-to-severe disease compared with conventional treatment (13.2 days vs 25.1 days; P < 0.05). The amelioration of Pediatric Ulcerative Colitis Activity Index score between before and 1 week after pulse steroid therapy was significantly more than that of conventional treatment (P < 0.01). No serious adverse effects were observed in the patients treated with pulse steroid therapy. However, the rate of the relapse episodes during the next 12 months after pulse steroid therapy was not significantly different from that after conventional treatment. CONCLUSION: These findings suggest that pulse steroid therapy is an option to be considered in children with moderate-to-severe UC. PMID- 21624005 TI - Cognitive P300-evoked potentials in school-age children after surgical or transcatheter intervention for ventricular septal defect. AB - BACKGROUND: Some studies have suggested that neurological development may be adversely affected in children with severe coronary heart disease who have undergone long periods of deep hypothermic cardiopulmonary bypass (CPB). Reports of cognitive function in VSD patients in whom surgical repair required only a relatively brief period of CPB are rare. Also, CPB is unnecessary for VSD patients undergoing transcatheter closure. The aim of this study was to assess the cognitive function in patients with ventricular septal defect. METHODS: A total of 29 patients treated with surgery, and 35 treated with transcatheter closure and their age- and sex-matched best friends completed the cognitive P300 auditory-evoked potentials test and the intelligence test. RESULTS: The patients and their best friends had normal intelligence quotient; however, the patients had longer P300 peak latencies in cranial frontal lobe and cranial vertex leads (329.2 +/- 24.8 and 335.1 +/- 20.0 ms) than the healthy controls did (319.1 +/- 20.6 and 313 +/- 18.2 ms) (P < 0.05). Patients who underwent surgery had longer P300 peak latency in the cranial frontal lobe and cranial vertex leads than did those with transcatheter closure and controls. When cardiopulmonary bypass and aortic clamping were used, the duration was associated with P300 peak latency for patients (P < 0.05). CONCLUSION: VSD patients, especially those undergoing surgery, showed poor cognitive function, which may be associated with duration of cardiopulmonary bypass or aortic-clamping. PMID- 21624006 TI - Phase I and II study of azacitidine in Japanese patients with myelodysplastic syndromes. AB - Azacitidine, an inhibitor of DNA methyltransferase, is reported to have antileukemic efficacy and is approved for the treatment of myelodysplastic syndromes in Western countries. We have conducted a Phase I/II study of azacitidine in Japanese patients with myelodysplastic syndromes to evaluate its pharmacokinetics, efficacy, and safety. In all, 53 patients received 75 mg/m(2) azacitidine subcutaneously or intravenously once daily for seven consecutive days on a 28-day cycle. The C(max) following intravenous administration was approximately 3.7-fold higher than that following subcutaneous administration, whereas the area under the plasma concentration-time curve from time zero to infinity was comparable for subcutaneous and intravenous administration. The bioavailability of azacitidine following subcutaneous administration was 91.1%, indicating that azacitidine is nearly completely absorbed after subcutaneous administration. The hematologic improvement and hematologic response rates were 54.9% (28/51) and 28.3% (15/53), respectively, and there were no differences between the two routes of administration. Azacitidine was generally well tolerated and clinically manageable in Japanese patients with myelodysplastic syndromes. Adverse events occurred in >= 20% of patients included hematologic toxicity, gastrointestinal events, and general disorders, such as malaise. Grade 3/4 adverse events that occurred in >= 50% of patients were all due to hematologic toxicity. The safety profile of azacitidine was generally similar for both routes of administration, with the exception of injection site reactions observed following subcutaneous administration. These results indicate that azacitidine can be expected to be a useful therapeutic agent in Japanese patients with myelodysplastic syndromes. PMID- 21624007 TI - Feasibility and pharmacokinetic study of bendamustine hydrochloride in combination with rituximab in relapsed or refractory aggressive B cell non Hodgkin's lymphoma. AB - Although bendamustine plus rituximab has demonstrated efficacy in indolent B cell non-Hodgkin's lymphoma (B-NHL), data for this combination in aggressive B-NHL are extremely limited. The present dose-escalation study evaluated the safety, efficacy, and pharmacokinetics of bendamustine hydrochloride in combination with rituximab in patients with relapsed/refractory, CD20-positive, aggressive B-NHL. Patients received rituximab 375 mg/m(2) , i.v., on Day 1 and bendamustine at either 90 (Cohort 1) or 120 mg/m(2) (Cohort 2), i.v., on Days 2 and 3 of a 21-day cycle. The primary endpoint was the proportion of patients experiencing dose limiting toxicity (DLT). Secondary endpoints were adverse events (AE), the overall response rate (ORR), and pharmacokinetic parameters. Nine patients received rituximab plus bendamustine: three in Cohort 1 and six in Cohort 2. Histologies included diffuse large B cell lymphoma (n = 5), mantle cell lymphoma (n = 2), and transformed lymphoma (n = 2). No DLT was observed at either dose level. Grade 3/4 hematologic AE included lymphocytopenia, leukocytopenia, and neutropenia (n = 9 each; 100%), and thrombocytopenia (n = 2; 22%). No Grade 3/4 gastrointestinal AE were reported. The ORR was 33% (one partial response) in Cohort 1 and 100% (five complete and one partial response) in Cohort 2. The maximum drug concentration and area under the blood concentration-time curve for bendamustine increased dose dependently, with time to maximum blood concentration = 1.0 h in both cohorts; these pharmacokinetic data were similar to those reported previously for single-agent bendamustine in patients with indolent B NHL. In conclusion, bendamustine 120 mg/m(2) plus rituximab 375 mg/m(2) was feasible and generally well tolerated, with promising efficacy in relapsed or refractory aggressive B-NHL. PMID- 21624008 TI - Aberrations of NEGR1 on 1p31 and MYEOV on 11q13 in neuroblastoma. AB - MYEOV and NEGR1 are novel candidate gene targets in neuroblastoma that were identified by chromosomal gain in 11q13 and loss in 1p31, respectively, through single nucleotide polymorphism array analysis. In the present study, to assess the involvement of MYEOV and NEGR1 in the pathogenesis of neuroblastoma, we analyzed their mutation status and/or expression profiles in a panel of 55 neuroblastoma samples, including 25 cell lines, followed by additional functional studies. No tumor-specific mutations of MYEOV or NEGR1 were identified in our case series. Expression of MYEOV was upregulated in 11 of 25 cell lines (44%) and in seven of 20 fresh tumors (35%). The siRNA-mediated knockdown of MYEOV in NB-19 cells, which exhibit high expression of MYEOV, resulted in a significant decrease in cell proliferation (P = 0.0027). Conversely, expression studies of NEGR1 revealed significantly lower expression of this gene in neuroblastomas at an advanced stage of the disease. Exogenous NEGR1 expression in neuroblastoma cells induced significant inhibition of cell growth (P = 0.019). The results of these studies provide supporting evidence for MYEOV and NEGR1 as gene targets of 11q13 gains and 1p31 deletions in a neuroblastoma subset. In addition, the findings suggest a possible prognostic value for NEGR1 in neuroblastoma. PMID- 21624009 TI - Risk factors for breast cancer: epidemiological evidence from Japanese studies. AB - Although our understanding of the etiology of breast cancer has improved, many well-known risk factors are not modifiable and present knowledge has proved insufficient to allow the disease to be overcome. Indeed, incidence and mortality among Japanese women have increased over the past three decades. Here, we review epidemiological evidence from our cohort and case-control studies among Japanese women in comparison with other published findings. Our studies confirm the important role of established factors derived primarily from Western populations, such as menstrual and reproductive factors, anthropometric factors, physical activity, and alcohol intake, in the development of breast cancer. In addition, we provide further evidence to better understand the role of traditional Japanese foods in the etiology of breast cancer. Our cohort study found that a higher intake of isoflavone and higher levels of plasma genistein, but not daidzein, were associated with a decreased risk of breast cancer. Our case-control studies reveal a dose-response pattern for these compounds; specifically, decreased risk as women move from "no" to "moderate" intake and leveling off thereafter. In addition, gene-environment interactions have been revealed in the effects of isoflavones. The evidence reviewed suggests that isoflavone has a protective effect against breast cancer in Asian populations. Conversely, our cohort study did not observe an inverse association between breast cancer risk and the intake of green tea and/or the plasma level of tea polyphenols, but we did find an association between increased risk and active and passive smoking. In conclusion, based on current knowledge, primary prevention according to individual lifestyle modification should focus on alcohol intake, weight control, physical activity, and tobacco smoking. PMID- 21624010 TI - Non-invasive, dynamic imaging of murine intestinal motility. AB - BACKGROUND: After intravenous (i.v.) administration, indocyanine green (ICG) is known to be secreted into bile from the liver via the biliary tracts, enabling fluorescent delineation of the intestine. In addition, ICG is a near-infrared (NIR) excitable fluorophore, capable of providing exogenous contrast for rapid NIR fluorescence imaging. We sought to quantify the intestinal motility using dynamic NIR fluorescence imaging after injection of ICG. METHODS: C57BL6 mice were dynamically imaged immediately before and up to 24h after i.v. and intradermal (i.d.) injection of 50 and 10MUL of ICG, respectively. Necropsy was also performed 1h postinjection and the entire gastrointestinal tract was isolated and exposed for ex vivo fluorescence imaging. KEY RESULTS: The secretion of ICG-laden fluorescent bile into the duodenum was observed in vivo and confirmed in situ. Different patterns of the intestinal motility, such as peristaltic and segmental motions, were dynamically imaged in vivo. Our imaging data showed that the frequency of contractions ranged from 27 to 35cyclesmin(-1) and the propagation velocity of peristaltic waves ranged from 0.82+/-0.5 to 2.04+/-1.12cms(-1) . CONCLUSIONS & INFERENCES: Dynamic NIR fluorescence imaging with injection of ICG can provide a method for diagnostic motility testing for intestinal motility disorders or dysfunction and for potential evaluation of therapeutic agents. PMID- 21624011 TI - Successful eradication of inhibitor of late recurrence and other high risk prognostic factors in a patient with severe haemophilia A. PMID- 21624012 TI - A systematic review: The use of desmopressin for treatment and prophylaxis of bleeding disorders in pregnancy. AB - Desmopressin (DDAVP) is commonly used for treatment and prevention of bleeding complications in patients with bleeding disorders including haemophilia A, von Willebrand's disease (VWD) and other less common disorders. This article reviews the current evidence for the use of DDAVP in pregnancy to clarify its efficacy and safety with regard to maternal and foetal outcome. A search of the literature found 30 studies that reported DDAVP use in pregnancy for prophylaxis or treatment of bleeding complications with 216 pregnancies reported in total. The most common indication was prophylaxis for prevention of bleeding during pregnancy and postpartum haemorrhage. DDAVP was used successfully in the first and early second trimester for bleeding prophylaxis in 50 pregnancies. No postpartum bleeding complications were reported in 167 out of 172 pregnancies when DDAVP was used for peripartum haemostatic cover. Twenty-nine studies reported no significant adverse events as a result of treatment with DDAVP. One case of water intoxication seizure and one case of premature labour following the use of DDAVP was reported in a single study. Other maternal side effects included facial flushing and headache and were reported by one study. These side effects were generally well tolerated by patients. There were no other significant adverse events reported in any of the studies as a result of DDAVP use. Foetal outcome was recorded in ten studies with no adverse foetal outcomes. In conclusion, this review shows that DDAVP in selected cases is effective in reducing bleeding complications associated with pregnancy and childbirth with a good safety record. Further research is needed to confirm these findings as they are based on the currently available evidence from small studies and case series only. PMID- 21624013 TI - Association of clinicopathologic parameters with the expression of inducible nitric oxide synthase and vascular endothelial growth factor in mucoepidermoid carcinoma. AB - BACKGROUND: The roles that inducible nitric oxide synthase (iNOS) and vascular endothelial growth factor (VEGF) play in tumorigenesis have been given special attention. In many tumors, their expression is upregulated. In addition, iNOS can stimulate the expression of VEGF. This study was carried out to investigate the expression of iNOS and VEGF as well as their relationship with angiogenesis and the clinicopathological characteristics of mucoepidermoid carcinoma (MEC). METHOD: The expression of iNOS and VEGF was detected by Streptavidin-peroxidase immunohistochemistry, and microvessel density (MVD) was determined by anti-CD34 antibody staining in 70 MEC cases and 40 normal salivary gland tissues (NSG). Follow-up was performed on the 70 patients with MEC. Non-parametric tests were performed for the comparison of iNOS and VEGF expression. RESULTS: The positive expression rates of iNOS and VEGF were successively enhanced in NSG, well differentiated and poorly differentiated MEC (P < 0.05). MVD counts were positively correlated with the expression levels of iNOS and VEGF in MEC (P < 0.05). The expression of iNOS was positively correlated with the expression of VEGF (P < 0.05). iNOS and VEGF expression were significantly associated with tumor differentiation, size metastasis, and relapse (P < 0.05) but were not correlated lymph node metastasis and metastasis. CONCLUSION: Inducible nitric oxide synthase can stimulate the expression of VEGF, and their expression status may help assess tumor malignancy and patient prognosis. PMID- 21624014 TI - Multicentre prospective study on dengue classification in four South-east Asian and three Latin American countries. AB - OBJECTIVE: To evaluate the existing WHO dengue classification across all age groups and a wide geographical range and to develop a revised evidence-based classification that would better reflect clinical severity. METHODS: We followed suspected dengue cases daily in seven countries across South-east Asia and Latin America and then categorised them into one of three intervention groups describing disease severity according to the overall level of medical and nursing support required. Using a pre-defined analysis plan, we explored the clinical and laboratory profiles characteristic of these intervention categories and presented the most promising options for a revised classification scheme to an independent group of WHO dengue experts for consideration. Potential warning signs were also evaluated by comparing contemporaneous data of patients who progressed to severe disease with the data of those who did not. RESULTS: A total of 2259 patients were recruited during 2006-2007 and 230 (13%) of the 1734 laboratory-confirmed patients required major intervention. Applying the existing WHO system, 47/210 (22%) of patients with shock did not fulfil all the criteria for dengue haemorrhagic fever. However, no three-tier revision adequately described the different severity groups either. Inclusion of readily discernible complications (shock/severe vascular leakage and/or severe bleeding and/or severe organ dysfunction) was necessary to devise a system that identified patients requiring major intervention with sufficient sensitivity and specificity to be practically useful. Only a small number of subjects (5%) progressed to severe disease while under observation; several warning signs were identified, but much larger studies are necessary to fully characterize features associated with disease progression. CONCLUSIONS: Based on these results, a revised classification system comprised of two entities, 'Dengue' and 'Severe Dengue', was proposed and has now been incorporated into the new WHO guidelines. PMID- 21624015 TI - Monitoring drug effectiveness in kala-azar in Bihar, India: cost and feasibility of periodic random surveys vs. a health service-based reporting system. AB - OBJECTIVE: In 2009, a random survey was conducted in Muzaffarpur district to document the clinical outcomes of visceral leishmaniasis patients (VL) treated by the public health care system in 2008, to assess the effectiveness of miltefosine against VL. We analysed the operational feasibility and cost of such periodic random surveys as compared with health facility-based routine monitoring. METHODS: A random sample of 150 patients was drawn from registers kept at Primary Health Care centres. Patient records were examined, and the patients were located at their residence. Patients and physicians were interviewed with the help of two specifically designed questionnaires by a team of one supervisor, one physician and one field worker. Costs incurred during this survey were properly documented, and vehicle log books maintained for analysis. RESULTS: Hundred and 39 (76.7%) of the patients could be located. Eleven patients were not traceable. Per patient, follow-up cost was US$ 15.51 and on average 2.27 patients could be visited per team-day. Human resource involvement constituted 75% of the total cost whereas involvement of physician costs 51% of the total cost. CONCLUSION: A random survey to document clinical outcomes is costly and labour intensive but gives probably the most accurate information on drug effectiveness. A health service-based retrospective cohort reporting system modelled on the monitoring system developed by tuberculosis programmes could be a better alternative. Involvement of community health workers in such monitoring would offer the additional advantage of treatment supervision and support. PMID- 21624016 TI - Kinetics of UV(254) inactivation of selected viral pathogens in a static system. AB - AIMS: The objective of this study was to estimate UV(254) inactivation constants for four viral pathogens: influenza virus type A, porcine respiratory and reproductive syndrome virus (PRRSV), bovine viral diarrhoea virus (BVDV) and reovirus. METHODS AND RESULTS: Viruses in culture medium were exposed to one of nine doses of UV(254) and then titrated for infectious virus. Analysis showed that viral inactivation by UV(254) was more accurately described by a two-stage inactivation model vs a standard one-stage inactivation model. CONCLUSIONS: The results provided evidence for the existence of two heterogeneous viral subpopulations among the viruses tested, one highly susceptible to UV(254) inactivation and the other more resistant. Importantly, inactivation constants based on the one-stage inactivation model would have underestimated the UV(254) dose required for the inactivation of these viruses under the conditions of the experiment. SIGNIFICANCE AND IMPACT OF THE STUDY: To improve the accuracy of estimates, it is recommended that research involving the inactivation of micro organisms evaluates inactivation kinetics using both one-stage and two-stage models. These results will be of interest to persons responsible for microbial agents under laboratory or field conditions. PMID- 21624017 TI - Novel application of nitrifying bacterial consortia to ease ammonia toxicity in ornamental fish transport units: trials with zebrafish. AB - AIMS: To evaluate whether two commercial nitrifying bacterial consortia can function as biocontrol agents in ornamental fish transporting systems. METHODS AND RESULTS: The consortia were applied in a simulated set-up using zebrafish as the model organism in three trials. The efficacy of the bacterial consortia in controlling the ammonia level was validated by measuring water quality parameters such as total ammonia, nitrate and pH of the transport water. The bacterial community structure in the transport unit was studied using denaturing gradient gel electrophoresis. The consortia tested improved the nitrifying activity that in turn facilitated the reduction of ammonia that had accumulated during the transport. Bacterial profiles revealed the presence of both ammonia-oxidizing and nitrite-oxidizing bacteria in the transport bags. CONCLUSIONS: The application of the consortia during the transportation of zebrafish could profoundly improve the water quality by curbing ammonia accumulation. SIGNIFICANCE AND IMPACT OF THE STUDY: The potential of applying nitrifying bacteria as a bioremediation practice during the transport of ornamental fish has been demonstrated and this innovative approach contributes to the amelioration of current fish welfare in ornamental fish trade. PMID- 21624018 TI - Application of inorganic carrier-based formulations of fluorescent pseudomonads and Piriformospora indica on tomato plants and evaluation of their efficacy. AB - AIMS: Fluorescent pseudomonads are widely used as bioinoculants for improving plant growth and controlling phytopathogenic fungi. Piriformospora indica (Pi), a symbiotic root endophyte, also has beneficial effects on a number of plants. The present study focuses on the improvement of growth yields of tomato plants and control of Fusarium wilt using inorganic carrier-based formulations of two fluorescent pseudomonad strains (R62 and R81) and Pi. METHODS AND RESULTS: The inorganic carrier-based formulations of pseudomonad strains and Pi were tested for plant growth promotion of tomato plants under glass house and field conditions. In controlled glass house experiments, 8.8-fold increase in dry root weight and 8.6-fold increase in dry shoot weight were observed with talcum powder based consortium formulation of R81 and Pi. Field trial experiments ascertained the glfass house results with a considerable amount of increase in plant growth responses, and amongst all the treatments, R81 + Pi treatment performed consistently well in field conditions with an increase of 2.6-, 3.1- and 3.9-fold increase in dry root weight, shoot weight and fruit yield, respectively. The fluorescent pseudomonad R81 and Pi also acted as biocontrol agents, as their treatments could control the incidence of wilt disease caused by Fusarium oxysporum f.sp. lycopersici in tomato plants under glass house conditions. CONCLUSIONS: The culture broths of pseudomonads R62, R81 and Pi were successfully used for development of talcum- and vermiculite-based bioinoculant formulations. In controlled glasshouse experiments, the talcum-based bioinoculant formulations performed significantly better over vermiculite-based formulations. In field experiments the talcum-based consortium formulation of pseudomonad R81 and Pi was most effective. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggests that the formulations of pseudomonad strains (R62 and R81) and Pi can be used as bioinoculants for improving the productivity of tomato plants. The application of such formulations is a step forward towards sustainable agriculture. PMID- 21624019 TI - Usefulness of real-time PCR as a complementary tool to the monitoring of Legionella spp. and Legionella pneumophila by culture in industrial cooling systems. AB - AIMS: This study was designed to evaluate the usefulness of quantification by real-time PCR as a management tool to monitor concentrations of Legionella spp. and Legionella pneumophila in industrial cooling systems and its ability to anticipate culture trends by the French standard method (AFNOR T90-431). METHODS AND RESULTS: Quantifications of Legionella bacteria were achieved by both methods on samples from nine cooling systems with different water qualities. Proportion of positive samples for L. pneumophila quantified by PCR was clearly lower in deionized or river waters submitted to a biocide treatment than in raw river waters, while positive samples for Legionella spp. were quantified for almost all the samples. For some samples containing PCR inhibitors, high quantification limits (up to 4.80 * 10(5) GU l(-1) ) did not allow us to quantify L. pneumophila, when they were quantified by culture. Finally, the monitoring of concentrations of L. pneumophila by both methods showed similar trends for 57 100% of the samples. CONCLUSIONS: These results suggest that, if some methodological steps designed to reduce inhibitory problems and thus decrease the quantification limits, could be developed to quantify Legionella in complex waters, the real-time PCR could be a valuable complementary tool to monitor the evolution of L. pneumophila concentrations. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows the possibility of using real-time PCR to monitor L. pneumophila proliferations in cooling systems and the importance to adapt nucleic acid extraction and purification protocols to raw waters. PMID- 21624020 TI - Growth optimization of algae for biodiesel production. AB - AIMS: Algae are favourable as a biofuel source because of the potential high oil content and fast generation of biomass. However, one of the challenges for this technology is achieving high oil content while maintaining exponential or high growth of the organism. Introducing a two-stage reactor to optimize both growth and oil content of the algae could be a solution to this hurdle. The aim of this study was to determine the reactor design parameters of the first-stage reactor, which would optimize growth of two algal strains, Oocystis sp. and Amphora sp. METHODS AND RESULTS: Growth kinetics were monitored by in vivo fluorescence and correlated to dry mass for both cultures under several environmental conditions during exponential growth. Temperatures of 25 and 30 degrees C and light intensities of 150 and 80 MUmol m(-2) s(-1) provided the most robust growth for Oocystis sp. and Amphora sp., respectively. Both strains showed optimized growth at a light : dark cycle of 16 : 08. At these conditions, the doubling rate for Oocystis sp. was 0.333 d(-1) and for Amphora sp. was 0.179 d(-1) . CONCLUSIONS: For both cultures, growth rate was more dependent on light : dark cycle and temperature than light intensity. Both strains grew slower in this work than data reported in the literature, however agitation and air/CO(2) sparging were not incorporated in the system under study. The highest doubling rate for Amphora sp. was observed near the maximum tolerable temperature, and it is suggested to grow this strain at 30 degrees C for a consistent high growth rate. SIGNIFICANCE AND IMPACT OF STUDY: Optimized growth conditions were determined for two lipid producing strains identified in the Aquatic Species Program summary report. An optimized, first-stage growth reactor operating at these conditions would thus offer the maximum productivity for an algal biomass feed stream into a lipid optimized second-stage reactor. PMID- 21624021 TI - Effects of environmental factors on microbial induced calcium carbonate precipitation. AB - AIMS: To gain an understanding of the environmental factors that affect the growth of the bacterium Sporosarcina pasteurii, the metabolism of the bacterium and the calcium carbonate precipitation induced by this bacterium to optimally implement the biological treatment process, microbial induced calcium carbonate precipitation (MICP), in situ. METHODS AND RESULTS: Soil column and batch tests were used to assess the effect of likely subsurface environmental factors on the MICP treatment process. Microbial growth and mineral precipitation were evaluated in freshwater and seawater. Environmental conditions that may influence the ureolytic activity of the bacteria, such as ammonium concentration and oxygen availability, as well as the ureolytic activities of viable and lysed cells were assessed. Treatment formulation and injection rate, as well as soil particle characteristics are other factors that were evaluated for impact on uniform induction of cementation within the soils. CONCLUSIONS: The results of the study presented herein indicate that the biological treatment process is equally robust over a wide range of soil types, concentrations of ammonium chloride and salinities ranging from distilled water to full seawater; on the time scale of an hour, it is not diminished by the absence of oxygen or lysis of cells containing the urease enzyme. SIGNIFICANCE AND IMPACT OF STUDY: This study advances the biological treatment process MICP towards field implementation by addressing key environmental hurdles faced with during the upscaling process. PMID- 21624022 TI - Imagined futures: how experiential knowledge of disability affects parents' decision making about fetal abnormality. AB - BACKGROUND: Knowledge of disability is considered key information to enable informed antenatal screening decisions by expectant parents. However, little is known about the role of experiential knowledge of disability in decisions to terminate or continue with a pregnancy diagnosed with a fetal abnormality. OBJECTIVE: To explore the role that expectant parents' experiential knowledge of disabilities and conditions can play in real-life decisions to continue or end a pregnancy with a fetal abnormality. DESIGN: Secondary analysis of qualitative narrative interview data informed by contextual systems framework. SETTING: Participants were recruited throughout the United Kingdom and interviewed between 2004 and 2006. PARTICIPANTS: Twenty-four women and four of their male partners who had direct or indirect experience of disability or illness and who had proceeded with or ended a pregnancy diagnosed with a fetal abnormality. FINDINGS: Most respondents recounted using their experiential knowledge of disability, whether of their unborn baby's condition or of a different condition, to try to imagine the future for their unborn child, themselves and their family when making their decision. Some, who were considering continuing their pregnancy and had little or no experience of their unborn baby's specific disability, sought out others' experiences of the condition following antenatal diagnosis.The nature of a parent's experiential knowledge did not predict whether they continued with or terminated their pregnancy. DISCUSSION: Prospective parents may find it helpful to discuss their existing knowledge of their unborn baby's condition with health professionals who are aware of the influence this might have on parents' decisions. PMID- 21624023 TI - Fathers' contributions to the management of their child's long-term medical condition: a narrative review of the literature. AB - CONTEXT: Fathers' contributions to the management of long-term childhood medical conditions are under-represented in the literature; therefore, the full extent of their involvement is poorly understood by practitioners and researchers, so strategies for promoting their involvement have not yet been fully considered. OBJECTIVE: To review studies of fathers' actual contributions in a wide range of conditions, the potential to optimize their contribution through additional interventions by health professionals and a direction for future research. DESIGN: Narrative review of the literature. METHODS: CINAHL, Medline, PsychInfo and ERIC databases were searched electronically between the years 1995-2008. The terms adherence, adjustment, child, chronic, compliance, concordance, condition, coping, disease, father, illness, information, long-term, management/intervention, mother, role, self-care and treatment were searched for separately and in combination. English language papers reporting primary research were selected and supplemented by hand-searching reference lists. Thirty-five papers (arising from 29 studies) met criteria and were selected for narrative review. RESULTS: Five themes were identified: (i) the impact of long-term conditions on fathers' ability to promote their child's well-being, (ii) factors influencing fathers' involvement in health care, (iii) personal growth/beneficial effects for fathers, (iv) the impact of father's involvement on family functioning and (v) strategies that increase fathers' participation in their child's health care and in research investigating fathers' participation. CONCLUSIONS: The review suggests that fathers' involvement in children's health care can positively impact on fathers', mothers' and children's well-being and family functioning. A range of strategies are identified to inform the promotion of fathers' contributions and future research investigating their input. PMID- 21624024 TI - How should patients behave to facilitate shared decision making--the doctors' view. AB - OBJECTIVE: To study how physicians feel about patients' efforts to be engaged in shared decision making (SDM). STUDY SETTING AND DESIGN: Survey of physicians from distinctly different medical disciplines (238 psychiatrists and 169 vascular surgeons). Participants were requested to judge which patient behaviours they find helpful and which behaviours detrimental for SDM. RESULTS: Psychiatrists and surgeons had rather positive attitudes about active patient behaviours. However, there were quite a few patient behaviours (e.g. searching the Internet, being assertive towards the doctor) which provoked ambivalent or negative attitudes. DISCUSSION AND CONCLUSIONS: Physicians are generally quite open towards active patient behaviour in the consultation. They, however, do consider it as less helpful and become more annoyed if patients insist on their preferences and doubt their doctors' recommendations. Physicians must realize that SDM implies giving up decisional power and try to be more flexible in their interactions with patients. PMID- 21624025 TI - Defining and measuring patient-centred care: an example from a mixed-methods systematic review of the stroke literature. AB - BACKGROUND: Involving patients in the determination of their care is increasingly important, and health-care professionals worldwide have recognized a need for clinical outcome measures and interventions that facilitate patient centred care delivery in a range of settings. AIM: A mixed-methods review was conducted, which aimed to identify stroke-specific patient-centred outcome measures and patient-centred interventions. SEARCH STRATEGY: Databases searched included MEDLINE and PsycINFO; search strings were based on MeSH terms and keywords associated with the terms 'stroke' and 'patient-centred'. DATA EXTRACTION AND ANALYSIS: Descriptive statistics were used to report quantitative data; thematic analysis was also performed in the included studies. MAIN RESULTS: Three patient-centred outcome measures (Subjective Index of Physical and Social Outcomes, Stroke Impact Scale, Communication Outcome after Stroke scale) and four interventions were identified. Key elements of intervention design included delivery in people's own homes, involvement of families and tailoring to individual needs and priorities. Thematic analysis enabled description of three broad themes: meaningfulness and relevance, quality, and communication, which informed the development of a definition of patient centred care specific to the specialty of stroke. CONCLUSIONS: It is important for health-care professionals to ensure that their practice is relevant to patients and families. The review identified three stroke-specific patient centred outcome measures, key elements of patient-centred interventions, and informed the development of a definition of patient-centred care. These review derived outputs represent a useful starting point for health-care professionals, whatever their specialty, who are working to reconcile tensions between priorities of health-care professionals and those of patients and their families, to ensure delivery of patient-centred care. PMID- 21624026 TI - Putting the 'patient' in patient safety: a qualitative study of consumer experiences. AB - BACKGROUND: Although patient safety has been studied extensively, little research has directly examined patient and family (consumer) perceptions. Evidence suggests that clinicians define safety differently from consumers, e.g. clinicians focus more on outcomes, whereas consumers may focus more on processes. Consumer perceptions of patient safety are important for several reasons. First, health-care policy leaders have been encouraging patients and families to take a proactive role in ensuring patient safety; therefore, an understanding of how patients define safety is needed. Second, consumer perceptions of safety could influence outcomes such as trust and satisfaction or compliance with treatment protocols. Finally, consumer perspectives could be an additional lens for viewing complex systems and processes for quality improvement efforts. OBJECTIVES: To qualitatively explore acute care consumer perceptions of patient safety. DESIGN AND METHODS: Thirty-nine individuals with a recent overnight hospital visit participated in one of four group interviews. Analysis followed an interpretive analytical approach. RESULTS: Three basic themes were identified: Communication, staffing issues and medication administration. Consumers associated care process problems, such as delays or lack of information, with safety rather than as service quality problems. Participants agreed that patients need family caregivers as advocates. CONCLUSIONS: Consumers seem acutely aware of care processes they believe pose risks to safety. Perceptual measures of patient safety and quality may help to identify areas where there are higher risks of preventable adverse events. PMID- 21624027 TI - (Un)organizing equal collaboration between users and professionals: on management of patient education in Norway. AB - BACKGROUND: This is an article about how patient education is managed in Norway, but it also addresses a matter of broader relevance that of how an organization imbued with a request for rational choices is able to take on board a contradictory ideology. In Norway, patient education under the auspice of hospitals is to be conducted as an equal collaboration between users and professionals, posing challenges to the ethos of rationally justified choices within the hospital sector. This calls for an exploration of how the organization copes with the contradictory demands. METHODS: A theoretical approach on the basis of theories from Scandinavian institutional theory and science and technology studies, informed by documents, interviews and experiences from national, regional and local levels in Norway. DISCUSSION: The field of patient education is divided into three decoupled domains: one at management level, one at the practical level, and in the middle a domain that acts as an interface between management and practice. This interface mediates the relationship between ideas and practice, without making overt the fact that ideas might not be possible to put into practice and that practice might not reflect ideas. CONCLUSIONS: The decoupling of practice and management allows patient education as equal collaboration between users and professionals to thrive as an idea, not subjugated by practical challenges. Thus, it can exist as a guiding star that both management and practitioners can attune to, but this situation might now be threatened by the demand for quality assurance in the field. PMID- 21624028 TI - Ecosystem services altered by human changes in the nitrogen cycle: a new perspective for US decision making. AB - Human alteration of the nitrogen (N) cycle has produced benefits for health and well-being, but excess N has altered many ecosystems and degraded air and water quality. US regulations mandate protection of the environment in terms that directly connect to ecosystem services. Here, we review the science quantifying effects of N on key ecosystem services, and compare the costs of N-related impacts or mitigation using the metric of cost per unit of N. Damage costs to the provision of clean air, reflected by impaired human respiratory health, are well characterized and fairly high (e.g. costs of ozone and particulate damages of $28 per kg NO(x)-N). Damage to services associated with productivity, biodiversity, recreation and clean water are less certain and although generally lower, these costs are quite variable (<$2.2-56 per kg N). In the current Chesapeake Bay restoration effort, for example, the collection of available damage costs clearly exceeds the projected abatement costs to reduce N loads to the Bay ($8-15 per kg N). Explicit consideration and accounting of effects on multiple ecosystem services provides decision-makers an integrated view of N sources, damages and abatement costs to address the significant challenges associated with reducing N pollution. PMID- 21624029 TI - Stochastic trajectories of succession initiated by extreme climatic events. AB - Deterministic or rule-based succession is expected under homogeneous biotic and abiotic starting conditions. Effects of extreme climatic events such as drought, however, may alter these assembly rules by adding stochastic elements. We monitored the succession of species composition of 30 twin grassland communities with identical biotic and abiotic starting conditions in an initially sown diversity gradient between 1 and 16 species over 13 years. The stochasticity of succession, measured as the synchrony in the development of the species compositions of the twin plots, was strongly altered by the extreme warm and dry summer of 2003. Moreover, it was independent from past and present plant diversity and neighbourhood species compositions. Extreme climatic events can induce stochastic effects in community development and therefore impair predictability even under homogeneous abiotic conditions. Stochastic events may result in lasting shifts of community composition, as well as adverse and unforeseeable effects on the stability of ecological services. PMID- 21624030 TI - Promastigote to amastigote differentiation of Leishmania is markedly delayed in the absence of PERK eIF2alpha kinase-dependent eIF2alpha phosphorylation. AB - The parasitic protozoan Leishmania is the etiological agent of human leishmaniasis worldwide. It undergoes cellular differentiation from the sandfly promastigote form into amastigotes within mammalian macrophages, a process that is essential for its intracellular survival. Here, we characterized the Leishmania infantum PERK eIF2alpha kinase homologue and addressed its role in the parasite's cytodifferentiation. We show that Leishmania PERK is an endoplasmic reticulum (ER) transmembrane protein that largely colocalizes with the ER BiP chaperone. The Leishmania PERK catalytic kinase domain undergoes autohyperphosphorylation and phosphorylates the translation initiation factor 2 alpha subunit (eIF2alpha) in vitro at threonine 166. We also report that PERK is post-translationally regulated specifically in the intracellular stage of the parasite or under ER stress, most likely through extensive autohyperphosphorylation. We have generated a PERK dominant negative mutant overexpressing a truncated PERK protein lacking the N-terminal luminal domain and showed that this mutant is impaired in eIF2alpha phosphorylation in response to ER stress or during amastigote differentiation. Most importantly, we showed that lack of eIF2alpha phosphorylation markedly delays the Leishmania differentiation process towards amastigote forms both in parasites grown axenically or within macrophages. These data highlight the importance of PERK eIF2alpha kinase dependent eIF2alpha phosphorylation in the intracellular development of Leishmania. PMID- 21624031 TI - Human liver sinusoidal endothelial cells respond to interaction with Entamoeba histolytica by changes in morphology, integrin signalling and cell death. AB - Invasive infection with Entamoeba histolytica causes intestinal and hepatic amoebiasis. In liver, parasites cross the endothelial barrier before abscess formation in the parenchyma. We focussed on amoebae interactions with human hepatic endothelial cells, the latter potentially playing a dual role in the infection process: as a barrier and as modulators of host defence responses. We characterized early responses of a human liver sinusoidal endothelial cell line to virulent and virulence-attenuated E. histolytica. Within the first minutes human cells start to retract, enter into apoptosis and die. In the presence of virulent amoebae, expression of genes related to cell cycle, cell death and integrin-mediated adhesion signalling was modulated, and actin fibre, focal adhesion kinase and paxillin localizations changed. Effects of inhibitors and amoeba strains not expressing pathogenic factors amoebapore A and cysteine protease A5 indicated that cell death and cytoskeleton disorganization depend upon parasite adhesion and amoebic cysteine proteinase activities. The data establish a relation between cytotoxic effects of E. histolytica and altered human target cell adhesion and suggest that interference with adhesion signalling triggers endothelial cell retraction and death. Understanding the roles of integrin signalling in endothelial cells will provide clues to unravel host pathogen interactions during amoebic liver infection. PMID- 21624032 TI - Cardiovascular outcomes associated with a new once-weekly GLP-1 receptor agonist vs. traditional therapies for type 2 diabetes: a simulation analysis. AB - AIM: The effect of glucose-lowering agents on diabetes-related complications including cardiovascular (CV) events is of major importance. In the absence of a long-term study, we simulated such a trial using a mathematical model where subjects were given exenatide once-weekly (EQW), which has been shown to improve glycaemic control and reduce weight, systolic blood pressure (SBP) and lipids in patients with type 2 diabetes mellitus (T2DM). METHODS: Using the Archimedes Model, we followed a simulated population derived from individuals with T2DM in NHANES who were drug-naive or on oral agents only. We modelled the effects of four treatment strategies including standard care (SC, maintaining levels of control seen in NHANES), intensive glycaemic control (IGC, target HbA1c < 7% with conventional antidiabetic agents) and two versions of EQW added to SC: one with glycaemic and weight reduction only (EQW-1) and one with additional improvements in SBP and lipids (EQW-2). EQW strategies were derived from 52-week clinical trial data. Endpoints included macrovascular and microvascular outcomes. RESULTS: Simulated EQW treatment resulted in earlier benefit and 2-3 times greater relative reductions in major adverse CV events than IGC when compared to SC (6% relative reduction by year 20 for IGC vs. 12 and 17% for the EQW strategies). For microvascular complications, EQW showed comparable benefit to IGC for neuropathy but significantly greater impact on renal complications. CONCLUSIONS: This analysis shows that the novel drug EQW has the potential to greatly reduce CV events through its combined effects on glycaemia, weight and other CV risk factors. PMID- 21624033 TI - Introduction of the ZmDof1 gene into rice enhances carbon and nitrogen assimilation under low-nitrogen conditions. AB - The excessive application of nitrogen fertilizer to maximize crop yields causes negative environmental effects such as pollution and ecological imbalance. To overcome this problem, researchers have attempted to improve the nitrogen assimilation capacity of crops. Maize Dof1 (ZmDof1) is a plant-specific transcription factor shown to promote nitrogen assimilation in Arabidopsis thaliana (Arabidopsis) even under nitrogen-deficient conditions. The present study examines the effect of the introduction of the ZmDof1 gene on carbon and nitrogen assimilation in rice. ZmDof1 induced the expression of phosphoenolpyruvate carboxylase (PEPC) genes in transgenic rice plants and transactivated the PEPC promoters in protoplast transient assays, showing similar effects in rice as in Arabidopsis. Transgenic rice expressing ZmDof1 and grown in the presence of 360 MUm (nitrogen-sufficient) or 90 MUm (nitrogen-deficient) of nitrogen concentrations showed modulation of metabolite content and gene expression associated with the anaplerotic pathway for the TCA cycle, suggesting an increased carbon flow towards nitrogen assimilation. Furthermore, increases in carbon and nitrogen amounts per seedling were found in Dof1 rice grown under nitrogen-deficient conditions. Nitrogen deficiency also resulted in the predominant distribution of nitrogen to roots, accompanied by significant increases in root biomass and modification of the shoot-to-root ratio. Measurement of the CO2 gas exchange rate showed a significant increase in the net photosynthesis rate in Dof1 rice under nitrogen-deficient conditions. Taken these together, the present study displayed that ZmDof1 expression in rice could induce gene expressions such as PEPC genes, modulate carbon and nitrogen metabolites, increase nitrogen assimilation and enhance growth under low-nitrogen conditions. PMID- 21624034 TI - Incidence of cervical intraepithelial neoplasia grade 2 or worse in colposcopy negative/human papillomavirus-positive women with low-grade cytological abnormalities. AB - OBJECTIVE: To determine the risk of incident high-grade cervical intraepithelial neoplasia (CIN) in human papillomavirus (HPV) -positive women with low-grade cytological abnormalities who had a satisfactory normal colposcopy. DESIGN: A retrospective follow-up study within the NHS HPV/LBC pilot studies. SETTING: The NHS Cervical Screening Programme in England. POPULATION: A total of 1063 HPV positive women with borderline or mild dyskaryosis who were negative at colposcopy from three sites within the NHS HPV/liquid-based cytology (LBC) pilot studies. METHODS: HPV triage took place in 2001/02. In 2009 all information on additional management on HPV-positive/colposcopy-negative women was requested. The rate of disease following a negative colposcopy was calculated, and survival analysis was used to determine whether the grade of referral cytology impacted on risk of subsequent disease. Results were compared with those in women from the same population who had not been HPV triaged. MAIN OUTCOME MEASURES: Incident CIN2 or worse during follow up. RESULTS: Of 1063 eligible women 965 had documented follow up. The cumulative rate of CIN2+ at 3 years in these women was 4.4% (95% CI 4.0-7.0%); the median time from normal colposcopy to final result was 27 months. There was no significant increase in the risk of future disease associated with age or initial cytology result. CONCLUSIONS: The rate of subsequent high-grade CIN among colposcopically negative triaged women was sufficiently low to justify return to routine recall. PMID- 21624035 TI - Outcomes following hysterectomy or endometrial ablation for heavy menstrual bleeding: retrospective analysis of hospital episode statistics in Scotland. AB - OBJECTIVE: To determine the risk of further gynaecological surgery and gynaecological cancer following hysterectomy and endometrial ablation in women with heavy menstrual bleeding. DESIGN: Population-based retrospective cohort study. SETTING: Scottish hospitals between 1989 and 2006. Population or sample Scottish women treated with hysterectomy or endometrial ablation for heavy menstrual bleeding between 1989 and 2006. METHODS: Anonymised data collected by the Scottish Information Services Division were analysed using appropriate methods across the hysterectomy and endometrial ablation groups. Cox proportional hazards regression analysis was used to examine the survival experience for different surgical outcomes after adjustment for age, year of primary operation and Carstairs quintile. MAIN OUTCOME MEASURES: Further gynaecological surgery and gynaecological cancer in women. RESULTS: A total of 37,120 women had a hysterectomy, 11,299 women underwent endometrial ablation without a subsequent hysterectomy and 2779 women underwent endometrial ablation followed by a subsequent hysterectomy. The median (interquartile range) duration of follow-up was 11.6 years (7.9, 14.8) and 6.2 years (2.7, 10.8) in the hysterectomy and endometrial ablation (without hysterectomy) cohorts, respectively. Compared with women who underwent hysterectomy, those who underwent ablation were less likely to need pelvic floor repair [adjusted hazards ratio, 0.62; 95% confidence interval (95% CI), 0.50, 0.77] or tension-free vaginal tape surgery for stress urinary incontinence (adjusted hazards ratio, 0.55; 95% CI, 0.41, 0.74). Abdominal hysterectomy was associated with a lower chance than vaginal hysterectomy of pelvic floor repair surgery (hazards ratio, 0.54; 95% CI, 0.45, 0.64). Overall, the number of women diagnosed with cancer was small, the largest group being breast cancer (n = 584, 1.57% and n = 130, 1.15% in the hysterectomy and endometrial ablation groups respectively; adjusted hazards ratio, 1.14; 95% CI, 0.93-1.39). CONCLUSIONS: Hysterectomy is associated with a higher risk than endometrial ablation of surgery for pelvic floor repair and stress urinary incontinence. Surgery for pelvic floor prolapse is more common after vaginal than abdominal hysterectomy. PMID- 21624036 TI - Risk of adverse perinatal outcomes with antithyroid treatment during pregnancy: a nationwide population-based study. AB - OBJECTIVE: To compare, using two large nationwide population-based data sets, the risk of adverse pregnancy outcomes (low birthweight [LBW], preterm birth, small for gestational age [SGA] and congenital anomalies) among pregnant women with hyperthyroidism classified into three groups: receiving propylthiouracil (PTU) treatment during pregnancy, receiving methimazole/carbimazole (MMI) treatment, and no antithyroid treatment during pregnancy. DESIGN: A matched case-control study. SETTING: Taiwan. SAMPLE: A total of 2830 mothers with hyperthyroidism and 14,150 age-matched randomly selected mothers without hyperthyroidism were included. METHODS: Conditional logistic regression analyses were performed to examine the risk of adverse pregnancy outcomes (LBW, preterm birth, SGA and major congenital anomalies) among these three groups. MAIN OUTCOME MEASURES: LBW, preterm birth, SGA and major congenital anomalies. RESULTS: Women receiving PTU treatment during pregnancy had a higher risk of giving birth to LBW infants than those not receiving antithyroid treatment (odds ratio = 1.40; 95% CI 1.00-1.96), after adjusting for maternal education, anaemia, hyperlipidaemia, pregestational diabetes, pregestational hypertension, hyperemesis gravidarum and infant's gender and birth order. However, children of women receiving MMI treatment did not have increased risks of any adverse fetal outcome relative to mothers not receiving antithyroid treatment. CONCLUSIONS: Our study finds an increased risk of LBW among babies of mothers with hyperthyroidism receiving PTU treatment during pregnancy relative to untreated mothers with hyperthyroidism. PMID- 21624037 TI - Mitochondrial DNA copy number and function decrease with age in the short-lived fish Nothobranchius furzeri. AB - Among vertebrates that can be kept in captivity, the annual fish Nothobranchius furzeri possesses the shortest known lifespan. It also shows typical signs of aging and is therefore an ideal model to assess the role of different physiological and environmental parameters on aging and lifespan determination. Here, we used Nothobranchius furzeri to study whether aging is associated with mitochondrial DNA (mtDNA) alterations and changes in mitochondrial function. We sequenced the complete mitochondrial genome of N. furzeri and found an extended control region. Large-scale mtDNA deletions have been frequently described to accumulate in other organisms with age, but there was no evidence for the presence of detectable age-related mtDNA deletions in N. furzeri. However, mtDNA copy number significantly decreased with age in skeletal muscle, brain, liver, skin and dorsal fin. Consistent with this finding, expression of Pgc-1alpha that encodes a transcriptional coactivator of mitochondrial biogenesis and expression of Tfam and mtSsbp both encoding mtDNA binding factors was downregulated with age. The investigation of possible changes in mitochondrial function revealed that the content of respiratory chain complexes III and IV was reduced in skeletal muscle with age. In addition, ADP-stimulated and succinate-dependent respiration was decreased in mitochondria of old fish. These findings suggest that despite the short lifespan, aging in N. furzeri is associated with a decline in mtDNA copy number, the downregulation of mtDNA-associated genes and an impairment of mitochondrial function. PMID- 21624038 TI - Insulin-activated Akt rescues Abeta oxidative stress-induced cell death by orchestrating molecular trafficking. AB - Increasing evidence indicates that Alzheimer's disease, one of the most diffused aging pathologies, and diabetes may be related. Here, we demonstrate that insulin signalling protects LAN5 cells by amyloid-beta42 (Abeta)-induced toxicity. Abeta affects both activation of insulin receptors and the levels of phospho-Akt, a critical signalling molecule in this pathway. In contrast, oxidative stress induced by Abeta can be antagonized by active Akt that, in turn, inhibits Foxo3a, a pro-apoptotic transcription factor activated by reactive oxygen species generation. Insulin cascade protects against mitochondrial damage caused by Abeta treatment, restoring the mitochondrial membrane potential. Moreover, we show that the recovery of the organelle integrity recruits active Akt translocation to the mitochondrion. Here, it plays a role both by maintaining unimpaired the permeability transition pore through increase in HK-II levels and by blocking apoptosis through phosphorylation of Bad, coming from cytoplasm after Abeta stimulus. Together, these results indicate that the Akt survival signal antagonizes the Abeta cell death process by balancing the presence and modifications of common molecules in specific cellular environments. PMID- 21624039 TI - Anaphylactic reaction induced by a polysulfone/polyvinylpyrrolidone membrane in the 10th session of hemodialysis with the same dialyzer. AB - The majority of severe hypersensitivity reactions in hemodialysis patients have occurred due to sensitization to ethylene oxide or to nonbiocompatible membrane dialyzers. The use of polysulfone dialyzers rarely causes hypersensitivity reactions. In the present study, we describe a case of severe life-threatening reactions induced by polysulfone dialyzers (from different manufacturers subjected to a variety of sterilization methods), which occurred after 9 sessions of hemodialysis with the same prescription, exemplifying the complexity of such reactions. PMID- 21624040 TI - Elastase release during the hemodialysis procedure seems to induce changes in red blood cell membrane proteins. PMID- 21624041 TI - Adding citrates to bicarbonate concentrates in the preparation of citrate enriched hemodialyzates. PMID- 21624042 TI - Carotid-jugular arteriovenous fistula: a case report of an iatrogenic complication following internal jugular vein catheterization for hemodialysis access. AB - We report a case of diabetic end-stage renal disease patient who presented with a right common carotid artery jugular arteriovenous fistula as a complication of the insertion of a polyurethane double-lumen hemodialysis catheter into the right internal jugular vein .On physical examination of the neck, a pulsating mass with a palpable thrill and a bruit was noted in the right subclavicular region. The diagnosis was confirmed by color doppler ultrasonography of the neck and carotid angiography. The review of the literature suggests the occurrence of this complication as rather rare. The fistula was successfully repaired surgically. It is emphasized that while securing the access, a thorough physical examination with a special emphasis on seeking any neck swellings, thrill, and bruit along with routine use of vascular doppler for securing dialysis access is recommended. PMID- 21624043 TI - Comparison of B-type natriuretic peptide and NT proBNP as predictors of survival in patients on high-flux hemodialysis and hemodiafiltration. AB - End stage renal failure is associated with very high risk of cardiovascular disease. Serum levels of B-type natriuretic peptide (BNP) and NT proBNP reflect cardiovascular risk but it is unknown which of these peptides is a better predictor of survival in this population. BNP and NT proBNP levels and other relevant parameters were measured in 103 patients on high-flux hemodialysis (HD) and hemodiafiltration. Patients were followed for 4 years or until transplantation or death. Median BNP level was 262 pg/mL while the corresponding NT proBNP level was 362 pg/mL. Levels of these peptides were significantly lower in patients receiving hemodiafiltration than in those on high-flux HD. Only 1 of the 26 patients with normal NT proBNP died during follow-up while 3 of the 33 patients with normal BNP levels died in the same period. Both median BNP and NT proBNP levels were higher in those who died during follow-up than in those who survived 4 years. Cox Proportional Hazard models showed that both logBNP and log NT proBNP were independent predictors of survival. The area under the receiver operating characteristic curve was very similar for BNP and NT proBNP (0.779 vs. 0.781) for predicting 4-year survival. Net reclassification improvement analysis showed that adding NT proBNP to the baseline model lead to improved prediction of 4-year survival. BNP and NT proBNP levels were markedly elevated in HD patients and were highly predictive of survival. NT proBNP may have marginal advantage over BNP in predicting survival in this population. PMID- 21624044 TI - A case of end-stage renal disease initially manifested with visual loss caused by uremic optic neuropathy. AB - Uremic optic neuropathy (UON) is extremely rare, although sporadic cases have been reported. UON is sometimes regarded as a manifestation of uremic neuropathy. Here, we report a case of end-stage renal disease presenting as UON. A 22-year old male was transferred to our nephrology department due to azotemia. Sudden deterioration of his vision occurred 3 days before his transfer. The patient's blood pressure was 150/90 mmHg, and he had no symptoms or signs of uremia, except for the visual disturbance. Blood tests showed an elevated serum creatinine of 6.0 mg/dL and serum BUN of 53.6 mg/dL. Both kidneys were decreased in size on ultrasound. His best-corrected vision was 20/62.5 in both the eyes. Ophthalmoscopy revealed hyperemia, swelling of both optic nerve heads, and blurred margins of both optic disks. These findings are compatible with UON. The patient's visual acuity and visual field improved following hemodialysis and corticosteroid treatment. The swelling of the patient's optic disks was also resolved. The patient is currently undergoing hemodialysis with the goal of vision restoration. Uremic optic neuropathy should be considered when patients with advanced chronic kidney disease complain of deterioration of their vision. PMID- 21624045 TI - Successful treatment of carbamazepine poisoning with hemodialysis: a case report and review of the literature. AB - A 48-year-old man was brought to the emergency room after ingesting an unknown amount of carbamazepine. He was unconscious and not responding to the noxious stimuli. He was intubated and was placed on mechanical ventilation because of respiratory insufficiency. Primary detoxification was performed with a gastric lavage and charcoal instillation. His serum carbamazepine level was 25.6 mcg/mL at the time of admission. His computed tomography of the brain was normal. He was managed conservatively but there was no improvement in his neurological status in the next 24 hours. Serum carbamazepine level was repeated and reported to be 28.3 mcg/mL. As there was no improvement in his sensorium and the serum carbamazepine levels remained persistently high, extracorporeal removal of carbamazepine was attempted. As the facility to carry out hemoperfusion was not available immediately, the decision to initiate hemodialysis was taken. After 3 sessions of hemodialysis, his sensorium improved markedly and the carbamazepine level at this time was within the therapeutic range. He was discharged after psychiatry consultation and counseling. We review the literature regarding extra corporeal techniques for the removal of carbamazepine and discuss them in this article. PMID- 21624046 TI - Elevated levels of PAI-1 activity and t-PA antigen are associated with newly diagnosed abnormal glucose regulation in patients with ST-elevation myocardial infarction. AB - BACKGROUND: Both Type 2 diabetes and cardiovascular disease have been associated with enhanced coagulation and suppressed fibrinolysis. OBJECTIVES: To investigate a possible relationship between selected hemostatic variables and abnormal glucose regulation (AGR) in patients with acute ST-elevation myocardial infarction (STEMI) without known diabetes and to study changes in selected hemostatic variables from baseline to follow-up in STEMI patients with or without AGR. METHODS: Plasminogen activator inhibitor-1 (PAI-1) activity, tissue plasminogen activator (t-PA) antigen, prothrombin fragment 1+2 (F(1+2)) and von Willebrand factor (vWF) were measured in fasting blood samples from 199 STEMI patients 16.5 h (median time) after admission and 3 months later. All patients were classified into normal glucose regulation (NGR) or AGR based on an oral glucose tolerance test at follow-up, according to the WHO criteria. RESULTS: High PAI-1 activity (>= 75th percentile) measured in-hospital was associated with AGR (n = 49) with an adjusted odds ratio of 2.2 (95% confidence interval, 1.1, 4.4). In addition, high levels of t-PA antigen (>= 75th percentile) were associated with AGR (adjusted odds ratio, 3.5; 95% confidence inteval, 1.5, 8.2), but only in men. Changes in the levels of F(1+2) were significantly more pronounced in patients with AGR compared with NGR (adjusted P = 0.04). CONCLUSION: Elevated levels of PAI-1 activity and t-PA antigen measured in-hospital in STEMI patients were associated with AGR classified at 3-month follow-up. Additionally, changes in the levels of F(1+2) were more pronounced in patients with AGR compared with NGR. The data suggest an enhanced prothrombotic state after an acute STEMI in patients with AGR without known diabetes. PMID- 21624047 TI - Bleeding risk in 'real world' patients with atrial fibrillation: comparison of two established bleeding prediction schemes in a nationwide cohort. AB - BACKGROUND: Oral anticoagulation (OAC) in patients with atrial fibrillation (AF) is a double-edged sword, because it decreases the risk of stroke at the cost of an increased risk of bleeding. We compared the performance of a new bleeding prediction scheme, HAS-BLED, with an older bleeding prediction scheme, HEMORR(2)HAGES, in a cohort of 'real-world' AF patients. METHODS: By individual level-linkage of nationwide registers, we identified all patients (n = 118,584) discharged with non-valvular AF in Denmark during the period 1997-2006, with and without OAC. Major bleeding rates during 1 year of follow-up were determined, and the predictive capabilities of the two schemes were compared by c-statistics. The risk of bleeding associated with individual risk factors composing HAS-BLED was estimated using Cox proportional-hazard analyses. RESULTS: Of AF patients receiving OAC (n = 44,771), 34.8% and 47.3% were categorized as 'low bleeding risk' by HAS-BLED and HEMORR(2)HAGES, respectively, and the bleeding rates per 100 person-years were 2.66 (95% confidence interval [CI], 2.40-2.94) and 3.06 (2.83-3.32), respectively. C-statistics for the two schemes were 0.795 (0.759 0.829) and 0.771 (0.733-0.806), respectively. The risk factors composing HAS-BLED were associated with varying risks, with a history of bleeding (hazard ratio [HR] 2.98; 95% CI 2.68-3.31) and being elderly (HR 1.93; 95% CI 1.71-2.18) being associated with the highest risks. Comparable results were found in AF patients not receiving OAC (n = 77,813). CONCLUSIONS: In an unselected nationwide cohort of hospitalized patients with atrial fibrillation, the HAS-BLED score performs similarly to HEMORR(2)HAGES in predicting bleeding risk but HAS-BLED is much simpler and easier to use in everyday clinical practise. PMID- 21624049 TI - The prevalence of maxillofacial fractures due to domestic violence--a retrospective study in a hospital in Tehran, Iran. AB - BACKGROUND/AIM: Domestic violence has been identified as a cause of maxillofacial fractures especially among women. The present study investigated the maxillofacial fractures in Tehran, Iran, with special focus on injuries related to domestic violence. MATERIAL AND METHODS: Records of patients with maxillofacial fractures who were referred to Shariati hospital, Tehran, from June 2004 to June 2006 were considered to extract required data. The extracted data included the patients' gender, age, cause of fracture, and type of fracture. For patients with domestic violence recorded as their fracture cause, complementary data were also recorded. Chi-square test served for statistical analysis. RESULTS: Totally, the records of 257 patients (188 men, 69 women) were considered. Of the 257 records studied, 188 records (73%) belonged to men. The most common fracture cause was vehicle accidents (55.3%) among both men (52.1%) and women (63.8%). The domestic violence stood for 3.5% of cases (all women). It was the fracture cause among 13% of women. One-third of women with domestic violence as fracture cause had previous history of fracture. The spouses of one third of domestic violence victims were drug addicted. CONCLUSION: Prevalence of domestic violence as a cause for maxillofacial fracture is relatively high among women. Because these fractures can be life-threatening, appropriate strategies at both community and family levels should be implemented to prevent and reduce these types of fractures. PMID- 21624050 TI - Maternal nutrition in rural Kenya: health and socio-demographic determinants and its association with child nutrition. AB - High levels of food insecurity and human immunodeficiency virus (HIV) infection place most breastfeeding mothers in Kenya at high risk of malnutrition. We examined the role of selected socio-economic, demographic and health factors as determinants of nutritional status among HIV-infected and HIV-uninfected mothers in rural Kenya and further examined the interrelationship between maternal nutritional and child nutritional status within this population. A cross sectional design was used to collect data from non-pregnant mothers with children ages 4-24 months in Kisumu District, Kenya. Over 80% of the mothers were breastfeeding at the time of the study. Mean maternal body mass index (BMI) (21.60 +/- 3.15) and percent body fat (22.29 +/- 4.86) values were lower than among lactating mothers in other Sub-Sahara African countries. Maternal HIV status was not significantly associated with any of the maternal nutritional indicators assessed in the study. Breastfeeding, recent severe illness and having multiple children below 2 years of age were negatively associated with maternal nutritional status, while higher maternal age, socio-economic status and household food security were each positively associated with maternal nutritional status. Significant positive association was reported between maternal weight, height, BMI, mid-upper arm circumference (MUAC), body fat and fat-free mass estimates, and children's height-for-age, weight-for-age, weight-for-height and MUAC-for-age z-score. This analysis identifies determinants of maternal nutritional status in rural Kenya and highlights the importance of interventions that address malnutrition in both HIV-infected and HIV-uninfected mothers in rural Kenya. Significant association between maternal and child nutritional status stresses the importance of addressing maternal and young child nutritional status as interrelated factors. PMID- 21624051 TI - Insights into the Rrf2 repressor family--the structure of CymR, the global cysteine regulator of Bacillus subtilis. AB - The global regulator CymR represses the transcription of a large set of genes involved in cystine uptake and cysteine biosynthesis in Bacillus subtilis and Staphylococcus aureus. This repressor belongs to the widespread and poorly characterized Rrf2 family of regulators. The crystal structure of CymR from B. subtilis reveals a biologically active dimer, where each monomer folds into two tightly packed domains: a DNA-binding domain, which houses a winged helix-turn helix (wHTH) motif; and a long dimerization domain, which places the wHTH motifs at the extremes. This architecture explains how these small regulators can span 23-27-bp DNA targets. The wHTH motif of CymR resembles those of the GntR superfamily of regulators, such as FadR and HutC. Superimposing the FadR wHTH motifs bound to their DNA fragments onto the wHTH motifs of the CymR dimer structure suggests that the DNA target and/or the protein must undergo some conformational changes upon binding. The CymR structure also hints at a possible location of the Fe-S centre associated with several Rrf2-type regulators. PMID- 21624052 TI - Exchange of the H(CC) domain mediating double receptor recognition improves the pharmacodynamic properties of botulinum neurotoxin. AB - The four-domain structure of botulinum neurotoxins (BoNTs) reflects their multistep intoxication process. The high toxicity of BoNTs primarily results from specific binding and uptake into neurons mediated by their 50-kDa cell-binding fragment (H(C) ). X-ray crystallography data have revealed that the H(C) fragment consists of two domains of equal size, named the 25-kDa N-terminal half of H(C) (H(CN) ) and the 25-kDa C-terminal half of H(C) (H(CC) ). In recent years, the ganglioside-binding sites of all seven BoNT serotypes have been allocated to the H(CC) domain. For BoNT/A, BoNT/B and BoNT/G, the protein receptor-binding site has been also been localized to the H(CC) domain. Here, we demonstrate that the H(CC) serotype can modulate the affinity of the H(C) fragment for neuronal membranes as well as the potency of full-length BoNT by replacing the BoNT/A H(CC) domain with the BoNT/B H(CC) , BoNT/C H(CC) and BoNT/E H(CC) domains, which exhibit higher affinity for synaptosomes. Indeed, the hybrids H(C) AB and H(C) AC display a higher affinity than wild-type H(C) A. Furthermore, the potency of a BoNT/A-based full-length hybrid containing the H(CC) B domain (AAAB; letters represent the serotype origin of the four domains) was quadrupled as compared with wild-type BoNT/A. Analogously, exchange of the H(C) fragment (AABB) yielded a neurotoxin with four-fold higher potency. As BoNT/A and BoNT/B are extensively used to treat neurological disorders, thereby facing the problem of BoNT neutralizing antibody formation, a BoNT with increased potency would lower the repeatedly administered protein dosage while maintaining the clinical benefit. Such a lowered protein load will delay the onset of neurotoxin antibody formation in patients. PMID- 21624053 TI - Enzymatic actions of Pasteurella multocida toxin detected by monoclonal antibodies recognizing the deamidated alpha subunit of the heterotrimeric GTPase Gq. AB - Pasteurella multocida toxin (PMT) is a virulence factor responsible for the pathogenesis of some Pasteurellosis. PMT exerts its toxic effects through the activation of heterotrimeric GTPase (G(q), G(12/13) and G(i))-dependent pathways, by deamidating a glutamine residue in the alpha subunit of these GTPases. However, the enzymatic characteristics of PMT are yet to be analyzed in detail because the deamidation has only been observed in cell-based assays. In the present study, we developed rat monoclonal antibodies, specifically recognizing the deamidated Galpha(q), to detect the actions of PMT by immunological techniques such as western blotting. Using the monoclonal antibodies, we found that the toxin deamidated Galpha(q) only under reducing conditions. The C terminal region of PMT, C-PMT, was more active than the full-length PMT. The C3 domain possessing the enzyme core catalyzed the deamidation in vitro without any other domains. These results not only support previous observations on toxicity, but also provide insights into the enzymatic nature of PMT. In addition, we present several lines of evidence that Galpha(11), as well as Galpha(q), could be a substrate for PMT. PMID- 21624054 TI - Solution parameters modulating DNA binding specificity of the restriction endonuclease EcoRV. AB - The DNA binding stringency of restriction endonucleases is crucial for their proper function. The X-ray structures of the specific and non-cognate complexes of the restriction nuclease EcoRV are considerably different suggesting significant differences in the hydration and binding free energies. Nonetheless, the majority of studies performed at pH 7.5, optimal for enzymatic activity, have found a < 10-fold difference between EcoRV binding constants to the specific and nonspecific sequences in the absence of divalent ions. We used a recently developed self-cleavage assay to measure EcoRV-DNA competitive binding and to evaluate the influence of water activity, pH and salt concentration on the binding stringency of the enzyme in the absence of divalent ions. We find the enzyme can readily distinguish specific and nonspecific sequences. The relative specific-nonspecific binding constant increases strongly with increasing neutral solute concentration and with decreasing pH. The difference in number of associated waters between specific and nonspecific DNA-EcoRV complexes is consistent with the differences in the crystal structures. Despite the large pH dependence of the sequence specificity, the osmotic pressure dependence indicates little change in structure with pH. The large osmotic pressure dependence means that measurement of protein-DNA specificity in dilute solution cannot be directly applied to binding in the crowded environment of the cell. In addition to divalent ions, water activity and pH are key parameters that strongly modulate binding specificity of EcoRV. PMID- 21624055 TI - Unique modifications of translation elongation factors. AB - Covalent modifications of proteins often modulate their biological functions or change their subcellular location. Among the many known protein modifications, three are exceptional in that they only occur on single proteins: ethanolamine phosphoglycerol, diphthamide and hypusine. Remarkably, the corresponding proteins carrying these modifications, elongation factor 1A, elongation factor 2 and initiation factor 5A, are all involved in elongation steps of translation. For diphthamide and, in part, hypusine, functional essentiality has been demonstrated, whereas no functional role has been reported so far for ethanolamine phosphoglycerol. We review the biosynthesis, attachment and physiological roles of these unique protein modifications and discuss common and separate features of the target proteins, which represent essential proteins in all organisms. PMID- 21624056 TI - Unravelling the twists and turns of the serpinopathies. AB - Members of the serine protease inhibitor (serpin) superfamily are found in all branches of life and play an important role in the regulation of enzymes involved in proteolytic cascades. Mutants of the serpins result in a delay in folding, with unstable intermediates being cleared by endoplasmic reticulum-associated degradation. The remaining protein is either fully folded and secreted or retained as ordered polymers within the endoplasmic reticulum of the cell of synthesis. This results in a group of diseases termed the serpinopathies, which are typified by mutations of alpha(1)-antitrypsin and neuroserpin in association with cirrhosis and the dementia familial encephalopathy with neuroserpin inclusion bodies, respectively. Current evidence strongly suggests that polymers of mutants of alpha(1)-antitrypsin and neuroserpin are linked by the sequential insertion of the reactive loop of one molecule into beta-sheet A of another. The ordered structure of the polymers within the endoplasmic reticulum stimulates nuclear factor-kappa B by a pathway that is independent of the unfolded protein response. This chronic activation of nuclear factor-kappa B may contribute to the cell toxicity associated with mutations of the serpins. We review the pathobiology of the serpinopathies and the development of novel therapeutic strategies for treating the inclusions that cause disease. These include the use of small molecules to block polymerization, stimulation of autophagy to clear inclusions and stem cell technology to correct the underlying molecular defect. PMID- 21624057 TI - Raloxifene reduces blood pressure in hypertensive animals after ovarian hormone deprivation. AB - Raloxifene is a selective oestrogen receptor modulator that has been approved for the prevention and treatment of osteoporosis in post-menopausal women. Studies have revealed several effects of raloxifene on the cardiovascular system, which might contribute to the blood pressure regulatory mechanisms, particularly in the systemic arterial hypertension. Therefore, the aim of this study was to investigate the effects of raloxifene on the blood pressure, renal excretion of water and Na(+) and plasma nitrite/nitrate levels in 2-kidney-1-clip (2K1C) hypertensive female rats. The groups were as follows: hypertensive (2K1C), hypertensive ovariectomized (2K1C + OVX) and hypertensive ovariectomized treated with raloxifene (2K1C + OVX + R). Seven days after the surgery that produced menopause, 2K1C hypertension was produced in anaesthetized animals. Seven days after the clip application, the rats were put into metabolic cages to allow for the measurement of water ingestion and diuresis, and raloxifene was administered (2 mg/kg/day i.p., for 7 more days). We found a large reduction (p < 0.01) in mean arterial pressure (197 +/- 6 to 164 +/- 2 mmHg), an increase in renal excretion of sodium and water (p < 0.05) and an increase in plasma levels of nitrite/nitrate in 2K1C + OVX + R animals, when compared with the 2K1C (23.4 +/- 1 versus 14 +/- 0.5 nmol/mL; p < 0.01, respectively). These findings suggest that raloxifene exerted its antihypertensive effect, at least in part, by improving the renal excretion of sodium and water. PMID- 21624058 TI - Synergistic effects of irinotecan and flavonoids on Ehrlich ascites tumour bearing mice. AB - Swiss albino mice were given Ehrlich ascites tumour cells (1 * 10(6)) intraperitoneally. For survival analysis and tumour growth analysis, the mice were administered quercetin and naringin (100 mg/kg) daily for 3 consecutive days, beginning on the third day after intraperitoneal (i.p.) injection of Ehrlich ascites tumour cells (1 * 10(6)). Irinotecan was administered ip at a dose of 50 mg/kg on days 1, 13 and 19. For the analysis of cell types and differential count of cells present in the peritoneal cavity, peripheral whole blood leucocyte count and the comet assay, the mice were treated therapeutically with quercetin and naringin (100 mg/kg) and irinotecan (50 mg/kg) daily for 3 consecutive days beginning on third day after i.p. injection of Ehrlich ascites tumour cells (1 * 10(6)). We observed the synergistic anti-tumour effect expressed as the median survival time of mice treated with naringin in combination with irinotecan. All test components inhibited tumour growth and increased lifespan of mice except quercetin. The total number of cells present in the peritoneal cavity of mice significantly decreased in all treatments except quercetin. Single irinotecan and irinotecan combined with naringin had the highest DNA-damaging potential on peripheral blood leucocytes and lowest primary DNA damage, both in the kidney and liver cells as measured by the alkaline comet assay. Our results showed enhanced anti-tumour activity of irinotecan in combined treatment with flavonoids to reduce the deteriorating reaction of cytostatic drugs. PMID- 21624059 TI - Effects of fluoxetine on the oxidative status of peripheral blood leucocytes of restraint-stressed mice. AB - Emotional stress can be viewed as a cause of adverse circumstances that induces a wide range of biochemical and behavioural changes. Oxidative stress is a critical route of damage in various psychological stress-induced disorders such as depression. Antidepressants are widely prescribed to treat these conditions; however, no animal study has investigated the effect of selective serotonin reuptake inhibitors (SSRIs) on the levels of intracellular reactive oxygen species in peripheral blood leucocytes of stressed mice. In this study, mice were immobilized for a period of 6 hr. Fluoxetine (5 mg/kg of body-weight) was administered 30 min. before subjecting the animals to acute stress. The level of intracellular reactive oxygen species in leucocytes of the peripheral blood of stressed mice was investigated using a 2',7'-dichlorofluorescein diacetate probe, and the antioxidant response of fluoxetine was evaluated by superoxide dismutase, diaphorase, catalase and reduced glutathione. Our results show that restraint stress significantly increases the generation of reactive oxygen species in the peripheral defence cells. Treatment with fluoxetine partially reverses the adverse effects of stress. The improvement in cellular oxidative status may be an important mechanism underlying the protective pharmacological effects of fluoxetine, which are clinically observed in the treatment of depressive disorders. PMID- 21624060 TI - Comparison of reproductive toxicity caused by cisplatin and novel platinum-N heterocyclic carbene complex in male rats. AB - Cisplatin and other platinum complexes are important chemotherapeutic agents and useful in the treatment for several cancers such as prostate, ovarian and testis. However, severe side effects including reproductive toxicity of cisplatin and other platinum complex cause limitations in their clinical usage. In this context, we aimed to compare the damage in testis caused by cisplatin and a novel platinum-N-heterocyclic carbene complex (Pt-NHC). To this end, 35 Sprague-Dawley rats were divided randomly into five equal groups (n = 7 in each group). Cisplatin and Pt-NHC were intraperitoneally administered as a single dose of 5 mg/kg or 10 mg/kg, and the rats were then killed 10 days after this treatment. The testicular tissues and serum samples were taken from all rats for the determination of reproductive toxicity. The results showed that cisplatin and Pt NHC caused toxicity on the reproductive system via increased oxidative and histological damage, decreased serum testosterone levels and negatively altered sperm characteristics in a dose-dependent manner (p < 0.05). At the same dose levels, cisplatin generally caused lower toxicity on the reproductive system compared with Pt-NHC. In conclusion, these results suggest that Pt-NHC has more toxic effects on the male reproductive system than cisplatin, and in terms of clinical usage, Pt-NHC may be unsafe compared with cisplatin. PMID- 21624061 TI - Tumour necrosis factor alpha -308G/A polymorphism and risk of the four most frequent cancers: a meta-analysis. AB - The latest data show that breast, prostate, lung and colorectal cancer are the four most frequent cancers in both sexes worldwide. A number of molecular epidemiological studies have been conducted to examine the association between TNF alpha -308G/A and the risk of those cancers. However the results have been inconclusive or inconsistent. We then performed a meta-analysis to derive a precise estimation of this association. We carried out a comprehensive search in Medline, EMBASE, OVID and Chinese Biomedical Literature Database for studies using related keywords. The inclusion criteria were (i) in English or Chinese; (ii) case-control study on this association; (iii) provide usable genotype frequencies; and (iv) sufficient published data for estimating an odds ratio (OR) with 95% confidence interval (CI). ORs and 95% CIs were calculated to assess the strength of this association under homozygote comparison (AA vs GG), heterozygote comparison (GA vs GG), dominant (AA/GA vs GG) and recessive (AA vs GA/GG) genetic model comparison. Thirty case-control studies with a total number of 16,507 cases and 19,749 controls were selected for analysis. Overall, no significant association was found between this polymorphism and the risk of total four cancers (GA vs GG: OR=1.02, 95% CI=0.91-1.14, P=0.78). However, there was a significant association between this polymorphism and breast cancer risk in western populations (GA vs GG: OR=0.91, 95% CI=0.85-0.96, P=0.002). This meta analysis also revealed that this polymorphism was not associated with susceptibility to the other three cancers. PMID- 21624066 TI - Report on the symposium celebrating the 40th anniversary of chronic dialysis and kidney transplantation in Slovenia. PMID- 21624067 TI - Slovenian Renal Replacement Therapy Registry: excerpts from the 2008 Annual Report. AB - This report provides a summary of the 2008 Slovenian Renal Replacement Therapy Registry Annual Report. Data on renal replacement therapy (RRT) were obtained from 20 dialysis centers and one transplant center, referring 31 December 2008, with 100% response rate to individual patient questionnaires. Slovenia has a population of approximately two million. The total number of patients treated by RRT was 1967, i.e. 968 per million of population (pmp), a 3.7% increase compared to 2007. In total, 1343 (68.3%) were treated by hemodialysis, 105 (5.3%) by peritoneal dialysis, and 519 (26.4%) had a functioning kidney graft. A total of 235 incident patients, 116 pmp (at day one), started RRT: their median age was 67 years, 61.8% were men, and 28% were diabetics. Regarding hemodialysis patients, 77% were dialyzed in hospital and 23% in private centers, 48% were treated with on-line hemodiafiltration and 74% with ultrapure dialysis fluid. The median weekly duration of hemodialysis was 13.5 hours, median dry body weight 68 kg, mean blood flow 283 +/- 51 mL/min, and 9.2% were dialyzed using a single-needle mode. Vascular access was provided by a native arteriovenous fistula in 82%, a polytetrafluoroethylene graft in 6%, and a catheter in 12%. The crude death rate was 12.4% in dialysis patients, 1.4% in transplant recipients, and 9.4% in all RRT patients (both dialysis and transplant, incident patients at day 1 included). The longest survival on RRT is 38 years (with hemodialysis only). Slovenia has been a member of Eurotransplant since 2000; 52 kidney transplantations were performed in 2008, all from deceased donors. PMID- 21624068 TI - Association of pretransplant renal replacement therapy duration with outcome in kidney transplant recipients: a prevalent cohort study in Slovenia. AB - The effect of renal replacement therapy (RRT) duration on kidney transplant outcome is controversial. The aim of this study was to analyze the association between pretransplant RRT duration versus patient and graft survival. The study cohort included 445 recipients of a deceased-donor kidney transplant between January 2000 and December 2009. Pretransplant RRT duration as a continuous variable and divided into time categories was the risk factor of interest. Patient and death-censored graft survival were the outcomes. Survival since the onset of RRT was calculated to avoid lead-time bias. Median pretransplant RRT duration was 4.7 years. The duration of RRT was longer in 33 patients who died (median 6.8 vs. 4.6 years; P = 0.022) and 56 patients who lost their graft (5.7 vs. 4.6 years; P = 0.035). Pretransplant RRT duration, as a continuous variable, was associated with a non-significant increase in the risk of recipient death (hazard ratio [HR] 1.01 per year of RRT; P = 0.09) and death-censored graft loss (HR 1.02; P = 0.12). When RRT was studied as a categorical variable, the mortality risk reached statistical significance when the patient had been on RRT for more than 4.7 years (HR 2.12; P = 0.042). Pretransplant RRT duration was not associated with an increased risk for recipient death if patient survival was calculated since the onset of RRT (HR 0.98 per year; P = 0.21). This study suggests that a longer RRT duration negatively impacts on post-transplant patient and graft survival; however, when pretransplant patient survival is accounted for, RRT duration has no significant effect on patient outcome. PMID- 21624069 TI - Dialysis patients after kidney graft failure: Slovenian experience. AB - Increased mortality has been reported in patients starting dialysis after kidney graft failure. In this study we analyzed this subgroup of dialysis patients based on the data from the Slovenian Renal Replacement Therapy Registry. Patients starting dialysis after graft failure in the period between 2004 and 2008 were identified from the registry. Demographic, clinical and treatment data, as well as survival were compared to incident dialysis patients, who were on the waiting list or preparing for enrollment. There were 49 patients starting dialysis after 7.9 +/- 6.4 years spent with a functioning graft and a total of 13.7 +/- 7.4 years on renal replacement therapy. Their mean age was 48.3 +/- 11.0 years (vs. 48.2 +/- 13.9 years in incident patients, P = 0.96), 53% were male, and all were on hemodialysis. By the end of 2008, 8 (16%) patients had been re-transplanted (after a median of 27.5 months) and 11 (23%) had died (after a median of 1.4 months of dialysis). The cause of death was infection in five patients, a cardiovascular event in three, malignancy in two, and a cerebrovascular event in one patient. Deceased patients were significantly older, but similar to survivors in other parameters. Unadjusted one- and three-year survival rates after graft failure were both 77%, which was significantly worse than in incident patients (P < 0.001). To conclude, patients after graft failure have increased mortality in the first year after starting dialysis, but patients surviving the first year have good survival thereafter. Studies focusing on the early period after graft failure are necessary to improve outcomes. PMID- 21624070 TI - Dialysis patients refusing kidney transplantation: data from the Slovenian Renal Replacement Therapy Registry. AB - Kidney transplantation is considered the best renal replacement therapy (RRT) for patients with end-stage renal disease; nevertheless, some dialysis patients refuse to be transplanted. The aim of our registry-based, cross-sectional study was to compare kidney transplant candidates to dialysis patients refusing transplantation. Data were collected from the Slovenian Renal Replacement Therapy Registry database, as of 31 December 2008. Demographic and some RRT data were compared between the groups. There were 1448 dialysis patients, of whom 1343 were treated by hemodialysis and 105 by peritoneal dialysis (PD); 132 (9%) were on the waiting list for transplantation, 208 (14%) were preparing for enrollment (altogether 340 [23%] dialysis patients were kidney transplant candidates); 200 (13.7%) patients were reported to refuse transplantation, all <= 65 years of age; 345 (24%) were not enrolled due to medical contraindications, 482 (33%) due to age, and 82 (6%) due to other or unknown reasons. No significant difference was found in age, gender, or presence of diabetes between kidney transplant candidates vs. patients refusing transplantation (mean age 50.5 +/- 13.9 vs. 51.3 +/- 9.6 years, males 61% vs. 63%, diabetics 18% vs. 17%). The proportion of patients <= 65 years old who were refusing transplantation was 28% (187/661) for hemodialysis and 17% (13/79) for PD patients (P = 0.03). There is a considerable group of dialysis patients in Slovenia refusing kidney transplantation. Compared to the kidney transplant candidates, they are similar in age, gender and prevalence of diabetes. Patients treated by peritoneal dialysis refuse kidney transplantation less often than hemodialysis patients. PMID- 21624071 TI - Severe peritonitis in patients treated with peritoneal dialysis: a case series study. AB - Severe peritonitis causing death and/or technique termination (catheter explanted) is one of the most devastating complications of peritoneal dialysis (PD). The aim of this case series study was to reveal the predictors of risk and clinical characteristics of these cases. We included 38 patients with either peritonitis causing death (18 patients, 47%) or catheter removal (20 patients, 53%) in the period 1996-2006. Their last clinical, laboratory and peritoneal equilibration test data before the peritonitis episode and hospitalization data after the start of peritonitis were reviewed. Their median (range) age was 66 (25 85) years, 61% were male, and the median PD duration was 60 (1-144) months. Baseline C-reactive protein (17.5 +/- 19.1 mg/L) was substantially higher than in contemporary stable controls from our unit (3.5 +/- 4.2 mg/L, P = 0.002). For 14 patients (37%), this was their first episode, with a significantly lower mortality of 14% as opposed to 47% across the whole group (P = 0.002). Almost half the patients (42%) had a causative abdominal condition identified, such as diverticulitis or cholecystitis. Clinical and laboratory data at presentation were variable and not different according to survival. Non-surviving cases had a significantly larger proportion of fast transporters (83 vs. 45%, P = 0.03), a significantly lower estimate of daily protein intake (0.72 vs. 0.88 g/kg/day, P = 0.007), and a significantly higher proportion of non-Gram-positive causative microorganisms (72 vs. 40%, P = 0.019). The patients with severe peritonitis were characterized as older with a longer PD duration, and a higher baseline C reactive protein. Fast peritoneal transport, lower normalized protein catabolic rate, and non-Gram-positive causative bacteria were associated with mortality. PMID- 21624072 TI - Treatment of anemia with epoetin in kidney transplant recipients. AB - The aim of this study was to analyze the prevalence and efficacy of renal anemia treated with epoetin in maintenance kidney transplant recipients in Slovenia. By the end of 2009, 107 out of 537 patients (19.9%) had been treated with epoetin. A cohort of 49 patients (45.8%) were analyzed in detail: 11 patients received epoetin alpha, 18 epoetin beta, 10 darbepoetin alpha, and 10 patients received methoxy polyethylene glycol-epoetin beta. The median epoetin dose was 0.36 ug/kg body weight per week. The median serum laboratory parameters were as follows: hemoglobin 120 g/L, hematocrit 0.36, ferritin 332 ng/mL, transferrin saturation 34%, serum creatinine 145 umol/L, serum albumin 41 g/L, intact parathyroid hormone 79 ng/L, and C-reactive protein 3 mg/L. We concluded that the prevalence of renal anemia in kidney transplant recipients treated with epoetin was approximately 20%, and laboratory parameters suggested that the treatment of renal anemia in this study cohort was optimal. PMID- 21624073 TI - Simple cystatin C formula compared to sophisticated CKD-EPI formulas for estimation of glomerular filtration rate in the elderly. AB - Despite the fact that the serum creatinine level is notoriously unreliable for the estimation of glomerular filtration rate (GFR) in the elderly, the serum creatinine concentration and serum creatinine-based formulas, such as the Modification of Diet in Renal Disease study equation (MDRD) are the most commonly used markers to estimate GFR. Recently, serum cystatin C-based formulas, the newer creatinine formula (the Chronic Kidney Disease Epidemiology Collaboration formula (CKD-EPI creatinine formula), and an equation that uses both serum creatinine and cystatin C (CKD-EPI creatinine and cystatin formula) were proposed as new GFR markers. The aim of our study was to compare the MDRD formula, CKD-EPI creatinine formula, CKD-EPI creatinine and cystatin formula, and simple cystatin C formula (100/serum cystatin C) against (51) Cr-EDTA clearance in the elderly. A total of 317 adult Caucasian patients aged >65 years were enrolled. In each patient, (51) Cr-EDTA clearance, serum creatinine, and serum cystatin C were determined, and the GFR was calculated using the MDRD formula, CKD-EPI formulas, and simple cystatin C formula. Statistically significant correlations between (51) Cr-EDTA clearance and all formulas were found. In the receiver operating characteristic (ROC) curve analysis with a cut-off of GFR 45 mL/min/1.73 m(2), a higher diagnostic accuracy was achieved with the equation that uses both serum creatinine and cystatin C (CKD-EPI creatinine and cystatin formula) than the MDRD formula (P < 0.013) or CKD-EPI creatinine formula (P < 0.01), but it was not higher than that achieved for the simple cystatin C formula (P = 0.335). Bland and Altman analysis for the same cut-off value showed that the creatinine formulas underestimated and the simple cystatin C formula overestimated measured GFR. All equations lacked precision. The accuracy within 30% of estimated (51) Cr EDTA clearance values differ according to the stage of CKD. Analysis of the ability to correctly predict GFR below and above 45 mL/min/1.73 m(2) showed a high prediction for all formulas. Our results indicate that the simple cystatin C formula, which requires just one variable (serum cystatin C concentration), is a reliable marker of GFR in the elderly and comparable to the creatinine formulas, including the CKD-EPI formulas. PMID- 21624074 TI - Partial remission of resistant nephrotic syndrome after oral galactose therapy. AB - Focal segmental glomerulosclerosis is sometimes associated with a circulating permeability factor. It was proposed that this factor interacts with the sugars of the glycocalyx, and its high affinity for galactose was shown on the basis of chromatographic studies. Galactose inactivates it and seems to lead to its clearance from plasma. A toddler with a nephrotic syndrome resistant to corticosteroids was admitted. A renal biopsy revealed minimal change disease with deposition of immunoglobulin M. Immunosuppressive therapy with pulses of cyclophosphamide, low-dose combination immunosuppressive therapy, and later with mycophenolate mofetil failed to induce remission. A renal biopsy six years later showed transformation to FSGS. After unsuccessful treatment with monthly pulses of cyclophosphamide, we began therapy with tacrolimus, which showed no effect. After two months, we added oral galactose to tacrolimus for one month, after which proteinuria decreased by 50%. Seven months later, galactose was again added for six months, after which proteinuria remained below 2 g/24 h and the plasma albumin and cholesterol concentrations normalized. An adolescent girl with a nephrotic syndrome resistant to corticosteroids was admitted. A renal biopsy revealed mesangioproliferative glomerulonephritis with C1q nephropathy. Therapy with tacrolimus failed to induce remission. After six months, we added galactose for three months, which reduced proteinuria to 0.76 g/24 h. After the discontinuation of galactose therapy, proteinuria increased to 2.48 g/24 h, despite further treatment with tacrolimus. It seems that oral galactose at a dose of 0.2 g/kg twice a day could be a promising new and nontoxic therapy for the treatment of resistant nephrotic syndrome. PMID- 21624075 TI - Forty-eight-hour ambulatory blood pressure and carotid-femoral pulse wave velocity in hemodialysis patients. AB - Carotid-femoral pulse wave velocity (cfPWV) is an important predictor of cardiovascular events in the general population and also in hemodialysis (HD) patients. In the general population, cfPWV is strongly associated with age and blood pressure (BP). The best timing and method of BP measurement in HD patients is uncertain. Ambulatory blood pressure measurements (ABPM) have been used to better define the relationship between BP, target organ damage, and outcomes in HD patients. The aim of this study was to determine the possible association between cfPWV, cardiovascular risk factors, single BP measurements, and 48-hour ABPM in chronic HD patients. Thirty-three HD patients (22 men, 11 women) were included. After the end of the mid-week HD session, BP was measured, arterial stiffness was estimated by cfPWV, and 48-hour ABPM was performed. The mean systolic and diastolic BP readings before cfPWV measurement were 136/79 mmn Hg, and the mean 48-hour systolic and diastolic BP readings were 131/76 mm Hg. The mean and range of the cfPWV measurements were 8.31 +/- 2.35 m/s and 5.18-16.53 m/s, respectively. Using regression analysis, no association between cfPWV and BP before PWV measurements was found. A statistically significant correlation between cfPWV and 48-hour systolic and diastolic ABPM was found. Using multiple regression analysis (including age, sex, smoking, diabetes, body mass index, total cholesterol, low- and high-density lipoprotein cholesterol, triglycerides, C-reactive protein, albumin, phosphorus, calcium, and iPTH) 48-hour systolic (P < 0.001) and diastolic ABPM (P < 0.005) still remain significantly associated with cfPWV. Only 48-hour ABPM was associated with cfPWV in HD patients in our study. We found no relationship between cfPWV and other cardiovascular risk factors. PMID- 21624076 TI - Long-term citrate anticoagulation in chronic hemodialysis patients. AB - In some cases, long-term (>3 months) citrate anticoagulation is needed in maintenance hemodialysis patients due to a persistent bleeding risk. In this retrospective observational study, we present our experience and assess its safety and effects on mineral and bone disorder parameters. Sixteen patients (mean age 67 +/- 15 years) were treated with long-term citrate anticoagulation. The indications were: recurrent gastrointestinal bleeding in nine patients, heparin-induced thrombocytopenia, retroperitoneal hematoma, chronic subdural hematoma, proliferative diabetic retinopathy, vascular malformations in the brain in one patient, and others in two patients. Metabolic complications and intact parathyroid hormone (iPTH) were analyzed. Citrate anticoagulation was performed for 4 months to 6.3 years (median 12 months). Ionized calcium was stable during the procedures; hypocalcemia (<0.9 mmol/L) was rare (2.1% of procedures), and there was one case of severe symptomatic hypocalcemia. There were no clinically significant acid-base disturbances and no clotting problems. In the short term (1 3 months after starting citrate), the iPTH increased in 73% of patients (from 325 +/- 310 to 591 +/- 793 pg/L, P = 0.11, N = 11). In the long term (1-2 years), an increase in iPTH was observed in 3/6 patients. The time period (before/after starting citrate) was a significant predictor of iPTH using main-effects anova (P < 0.001). To conclude, long-term citrate anticoagulation in chronic hemodialysis patients is safe. Mild hypocalcemia during dialysis with citrate anticoagulation may contribute to a short- and long-term increase in iPTH in these patients. Further studies on long-term citrate anticoagulation are necessary. PMID- 21624077 TI - Citrate anticoagulation during post-dilution hemodiafiltration with a high cut off (Theralite) membrane. AB - Citrate anticoagulation has not yet been described for hemodiafiltration (HDF) with high cut-off (HCO) membranes, which can be used in the treatment of cast nephropathy secondary to multiple myeloma. A 57-year-old male patient with multiple myeloma and acute renal failure was treated with HDF using a HCO membrane (Theralite) each or every other day. Due to thrombocytopenia, citrate anticoagulation was done for the first 7 h, and anticoagulant-free HDF was performed for the last hour to avoid citrate accumulation. Magnesium, phosphate, and albumin were measured after 3, 6, and 8 h, and were replaced as necessary. Thirty-two post-dilution HDF procedures (8 h each, infusate 24 L) were performed with blood flow at 300-330 mL/h; sodium citrate 4% was infused at 300 mL/h and 1 mol/L calcium chloride was infused at a mean rate of 14.6 +/- 1.1 mL/h. Calcium free dialysate/infusate was used. Ionized calcium was stable (1.10 +/- 0.06 before and 1.08 +/- 0.06 mmol/L after HDF). Magnesium was stable (0.67 +/- 0.12 before and 0.68 +/- 0.05 mmol/L after HDF), with an average 390 +/- 180 mg per procedure, substituted orally. There was no metabolic alkalosis or hypernatremia after the procedures, and no significant clotting was noted. The total/ionized calcium ratio (1.87 +/- 0.22 before vs. 1.56 +/- 0.20 after 6 h) and the corrected/ionized calcium ratio (2.02 +/- 0.21 before vs. 1.88 +/- 0.27 after 6 h) decreased during HDF, indicating no citrate accumulation. Citrate anticoagulation was effectively performed during 8 h of HCO membrane HDF. There were no side effects of citrate anticoagulation, nor were any signs of citrate accumulation noted. PMID- 21624078 TI - High cut-off membrane hemodiafiltration in myoglobinuric acute renal failure: a case series. AB - Acute renal failure is a major complication of rhabdomyolysis. New membranes for hemodialysis have been developed with a high cut-off pore size allowing efficient removal of myoglobin. We report on six patients treated by hemodiafiltration with a high cut-off membrane (HCO-HDF) for myoglobinuric acute renal failure. Rhabdomyolysis was caused by infection in two patients, by a statin in one patient and a non-traumatic crush in another, and followed cardiovascular surgery in two others. Ten HCO-HDF procedures were performed. A high cut-off hemofilter was used, with citrate anticoagulation and postdilutional fluid substitution of 2 3 L/h, dialysate flow 500 mL/min, and blood flow within 250-300 mL/min. Albumin losses were replaced by infusion of human albumin solution, and the mean myoglobin reduction ratio was 77% (range, 62-89%). An excellent clearance of 81 mL/min (range 42-131 mL/min) was achieved. Nearly 5 g of myoglobin was removed into the dialysate collected in one of the procedures. A high rebound in serum myoglobin, on average to 244% of the post-procedure myoglobin level, was observed. The four patients alive at the time remained anuric for a week. Slow myoglobin elimination with a mean half-time of 39 h (range 19-59 h) was observed in that period. Highly efficient myoglobin removal by high cut-off membrane hemodiafiltration was demonstrated in our patients. Rapid redistribution from the extracellular fluid and sustained myoglobin release were suggested by the high rebound observed. Elimination of myoglobin within the body was shown in our study to occur slowly during the period of anuria. PMID- 21624079 TI - Vascular access in children on chronic hemodialysis: a Slovenian experience. AB - The aim of our study was to report our experience with arteriovenous fistulas (AVFs) and non-cuffed central venous catheters (CVCs) in children and adolescents with end-stage renal disease (ESRD) on hemodialysis (HD). The children with ESRD (18 years or younger) who were hemodialyzed at the Center of Dialysis and Transplantation, Children's Hospital, Ljubljana, in the period between December 1998 and December 2010 were included in our retrospective study. We recorded the data considering the CVCs and AVFs used for HD. Thirty-one children (13 females, 18 males) with ESRD received HD treatment. The mean patient age when HD was started was 13.3 +/- 3.4 years. Altogether, 35 AVFs were created, and the primary failure rate was 25.7% (9/35). The time to maturation was 4.0 +/- 2.5 months. The mean patency of the AVF was 42.5 +/- 51.9 months. Seventy-seven CVCs (non-cuffed) were inserted in the observation period; 89.6% of the CVCs were inserted in the jugular vein, and citrate locking was used in the interdialysis period. The CVCs were removed after 0.1-17.4 months (3.6 +/- 3.7 months). The incidence of bacteremia was 0.9 episodes per 1000 catheter days. The preferred vascular accesses for pediatric hemodialysis are native AVFs; however, a single lumen, non cuffed, citrate-locked CVC placed in a jugular vein can be acceptable as a long term vascular access when AVF cannot be constructed or used. PMID- 21624080 TI - Detection of dialysis access induced limb ischemia by infrared thermography in children. AB - High arteriovenous fistula (AV fistula) blood flow may impair distal limb perfusion and cause irreversible ischemic damage. Since tissue temperature reflects blood perfusion, we tried to assess distal blood flow using an infrared camera. We examined all 12 patients with an AV fistula in our dialysis unit. Seven were pediatric patients aged 11.0-18.9 years (mean 14.9 years) and five were adults aged 26.9-62.1 years (mean 38.6 years). Infrared thermal imaging (thermography) of their hands was performed after the completion of their regular dialysis sessions. In each patient, the spot temperature of each fingertip on both hands was assessed separately, with three measurements being performed for each measuring point. The mean spot temperature of all fingertips was calculated for each hand and the results compared. A statistically significant difference (P < 0.05) indicated distal perfusion insufficiency. Perfusion of the hands was also assessed by inspecting the visualized temperature distribution on the thermal image. Finally, we compared the results to the clinical findings in relevant patients. In 8/12 patients (66.7%), the mean spot temperature of the fingertips was statistically significantly lower on the fistula side (P < 0.05). Only 4/12 patients (33.3%) had clinical symptoms, and all were detected by thermography. Abnormal findings were more frequent in elderly patients. Although we realize that the diagnosis of steal syndrome is primarily clinical, thermography might be a safe, noninvasive, cheap tool for the timely detection of children at risk of developing symptoms of hand ischemia. PMID- 21624081 TI - Surgical thrombectomy of thrombosed arteriovenous grafts by interventional nephrologists. AB - The aim of our retrospective study was to present the success of surgical thrombectomy in acutely thrombosed, arteriovenous (AV), expanded polytetrafluoroethylene (ePTFE) grafts. Patients from hemodialysis centers in Slovenia were admitted to our dialysis center after acute thrombosis of their AV graft to undergo surgical thrombectomy under local anesthesia as an outpatient procedure. In 55 chronic hemodialysis patients, of whom 26 were men (47.3%), with a mean age of 63 +/- 13 years (range 35-84 years), and diabetes mellitus in 11 patients (20%), 59 ePTFE AV grafts thrombosed. A total of 129 thrombectomies were performed, on average 2.2 +/- 2.1 per graft (range 1-13). Primary patency (defined as the time from AV graft creation to first thrombosis) was 638 +/- 633 days (range 10-2586, median 418 days), secondary patency (defined as the time from first thrombectomy to abandonment) was 451 +/- 472 days (range 0-1994, median 305 days), and cumulative patency (defined as the time from creation to abandonment) was 1089 +/- 685 days (range 25-3020, median 1031 days). In 46 (78%) of the AV grafts, the first thrombectomy was successful. The secondary patency rates after 1, 2, 3, 4, and 5 years were 76%, 66%, 54%, 14%, and 14%, respectively (13/59 unsuccessfully thrombectomized grafts were excluded). Cumulative patency after 1, 2, 3, 4, and 5 years was 88.1%, 67.8%, 44.7%, 27.1%, and 16.9%, respectively. In conclusion, surgical thrombectomy after thrombosis of an AV graft in the arm or thigh, performed by interventional nephrologists and followed, if required, by angioplasty, significantly prolonged the patency of the majority of thrombosed AV grafts. PMID- 21624082 TI - Two single-lumen noncuffed catheters in the jugular vein as long-term vascular access: a preliminary report. AB - Two single-lumen, noncuffed catheters in the same jugular vein have been used as preferred vascular access in our hemodialysis (HD) and apheresis patients in past years. The aim of this retrospective study was to analyze the clinical outcome of such a vascular access and the reasons for catheter removal. In 129 adult patients, aged 69 +/- 13 years, 56% males, treated by HD (121 patients) or apheresis (8 patients), two single lumen, pre-curved 8 Fr catheters (Medcomp, Harleysville, PA, USA) inserted into the same jugular vein were used as vascular access between January 2009 and April 2010. The catheters were inserted into the left jugular vein in 21 patients, and into the right jugular vein in 108 patients. A 30% solution of trisodium-citrate was used as a locking solution, and 2% mupirocin ointment was routinely applied to the exit site. The catheters were removed in 86 patients after 1-288 days, median 17.5 days. In 74 patients, there was either no need for further dialysis or an arteriovenous fistula was constructed (17 patients). In 10 patients, wire exchange was performed for correction of a displaced functional catheter (after 6-201 days), and in only two patients the catheters were removed due to infection on days 10 and 184. The longest period of a catheter functioning without intervention was 387 days. Fifteen patients died with functional catheters left in place (duration 1-387 days). In four patients, the catheters were still functional at the time of analysis (duration 198-268 days). Another nine patients were transferred to other dialysis centers (after they had been followed up at our center lasting for 1-63 days), with no data on their outcome after transfer. Fifteen patients were lost to follow up after insertion. Two single-lumen, noncuffed catheters in the same jugular vein, locked with 30% citrate, seem to be a safe and long lasting method of vascular access for hemodialysis and apheresis in some patients, but further prospective studies are needed to evaluate the clinical outcome of this type of vascular access. PMID- 21624083 TI - Value of beta2-microglobulin in the serum of healthy subjects older than 40 years. AB - The aim of this research was to determine if the age of healthy subjects older than 40 years of age has an influence on the concentration of beta(2) microglobulin in the serum of subjects of different populations. We examined the values of beta(2) -microglobulin in the serum of 51 healthy subjects aged 40-86 years using the microparticle enzyme immunoassay AxSYM beta(2) -microglobulin test. The reference values of beta(2) -microglobulin according to the nonparametric statistical method is 0.95-2.73 mg/L. A correlation was found between beta(2) -microglobulin and age: 40-50 years (0.94-1.54 mg/L), 51-65 years (0.96-2.62 mg/L), and >65 years (1.13-2.84 mg/L). There was no significant statistical difference of beta(2) -microglobulin between genders (P > 0.05); however, there was a statistically significant difference between the concentration of beta(2) -microglobulin and the subjects' age. (Spearman's rank correlation coefficient rho = 0.66; P < 0.01). A direct correlation between age and the concentration of beta(2) -microglobulin was observed. This research is a contribution to determining reference values of beta(2) -microglobulin in subjects of different populations. PMID- 21624084 TI - Evaluation of a new method for measuring vascular access recirculation. AB - Together with Nikkiso in Shizuoka, Japan, we developed a new method for measuring the rate of vascular access recirculation by the blood volume monitor. This measurement is performed via a method of dilution that employs a marker produced by rapid ultrafiltration using a dialysis machine. In this paper, we evaluate the reliability and safety of this machine, in vitro and in vivo. The safety of this method was evaluated by investigating hemolysis after rapid ultrafiltration. The measurement of free hemoglobin, potassium and haptoglobin in the circulating blood were performed before and after rapid ultrafiltration. No data was found to indicate hemolysis in vivo, detected by an increase in potassium or a decrease in haptoglobin. Evaluation of reliability in an experimental system was also performed on an in vitro recirculation system at a rate of 0, 10, 25, 50, and 70%. Almost all of the measured rates were within +/- 10% of the theoretical rate. We performed 20 hemodialysis experiments with vascular access recirculation applying this monitor and simultaneous urea and creatinine dilution methods, which were the recommended standard measurements for vascular recirculation. In 53 measurements of standard vascular shunts with no postural change, differences of the results between the monitor and both dilution methods were only 4.0% and 3.2%, respectively. Regression analysis showed a significant and positive correlation between them (P < 0.0001). We conclude that this new method for measuring vascular access recirculation is applicable in terms of both accuracy and ease of operation. PMID- 21624085 TI - Removal of doripenem during hemodialysis and the optimum dosing regimen for patients undergoing hemodialysis. AB - The removal of doripenem by hemodialysis was studied in six hemodialysis patients. Following an intravenous drip infusion of 0.5 g of doripenem, plasma concentrations of the drug were measured. The decrease in drug concentrations in plasma was observed during various periods of non-hemodialysis, and hemodialysis accelerated the elimination of doripenem. For example, the calculated mean half life during hemodialysis was significantly shorter than that during non hemodialysis periods (P = 0.002). The calculated pharmacokinetic parameters indicated that the mean rate of decrease in plasma concentration due to hemodialysis alone was 56.12 +/- 8.11%. Upon obtaining these results and several pharmacokinetic parameters, we attempted to optimize the dosing regimen of doripenem for hemodialysis patients. We recommend the use of 0.25 g of doripenem once a day in patients infected with viable bacteria, and in patients who are infected with Pseudomonas aeruginosa, 0.5 g twice a day on the first day of administration, followed by 0.5 g once a day. PMID- 21624086 TI - Lessons from Japan's March 2011 earthquake regarding dialysis patients. PMID- 21624087 TI - Ocular tuberculosis in a patient undergoing hemodialysis therapy. PMID- 21624088 TI - Refractory acquired hemophilia: successful treatment by immunoadsorption with single-use columns. PMID- 21624090 TI - Comprehensive analysis of single- and multi-target activity cliffs formed by currently available bioactive compounds. AB - Activity cliffs are formed by structurally similar compounds having large potency differences. Their study is a focal point of SAR analysis. We present a first systematic survey of single- and multitarget activity cliffs contained in currently available bioactive compounds. Approximately 12% of all active compounds were involved in the formation of activity cliffs. Perhaps unexpectedly, activity cliffs were found to be similarly distributed over different protein target families. Moreover, only approximately 5% of all activity cliffs were multitarget cliffs. Importantly, we also found that only very few multitarget cliffs were formed by compounds having different target selectivity. In addition, 'polypharmacological cliffs', i.e., multitarget activity cliffs involving targets from different protein families, were also only rarely found. Taken together, our findings reveal that only approximately 2% of all pairs of structurally similar compounds sharing the same biological activity form activity cliffs but that, on average, approximately one of 10 active compounds is involved in the formation of one or two single-target cliffs of large magnitude (with at least 100-fold difference in potency). These compounds provide a rich source of SAR information and can be identified across many different target families. PMID- 21624091 TI - Knowledge-based identification of the ERK2/STAT3 signal pathway as a therapeutic target for type 2 diabetes and drug discovery. AB - Many existing agents for diabetes therapy are unable to restore or maintain normal glucose homeostasis or prevent the eventual emergence of hyperglycemia related complication. Therefore, agents based on novel mechanisms are sought to complement and extend the current therapeutic approaches. Based on the initial paper research, we focused on active STAT3 as an attractive pharmacological target for type 2 diabetes. The subsequent text mining with a unique query to identify suppressors but not activators of STAT3 revealed the ERK2/STAT3 pathway as a novel diabetes target. The description of ERK2 inhibitors as diabetes target had not been found in our text mining research at present. The mechanism-based peptide inhibitor for ERK2 was identified using the knowledge of the KIM sequence, which has an important role in the recognition of cognate kinases, phosphatases, scaffold proteins, and substrates. The peptide inhibitor was confirmed to exert effects in vitro and in vivo. The peptide inhibitor conferred a significant decrease in HOMA-IR levels on Day 28 compared with that in the vehicle group. Besides lowering the fasting blood glucose level, the peptide inhibitor also attenuated the blood glucose increment in the fed state, as compared with the vehicle group. PMID- 21624092 TI - How do they do Wnt they do?: regulation of transcription by the Wnt/beta-catenin pathway. AB - Wnt/beta-catenin signalling is known to play many roles in metazoan development and tissue homeostasis. Misregulation of the pathway has also been linked to many human diseases. In this review, specific aspects of the pathway's involvement in these processes are discussed, with an emphasis on how Wnt/beta-catenin signalling regulates gene expression in a cell and temporally specific manner. The T-cell factor (TCF) family of transcription factors, which mediate a large portion of Wnt/beta-catenin signalling, will be discussed in detail. Invertebrates contain a single TCF gene that contains two DNA-binding domains, the high mobility group (HMG) domain and the C-clamp, which increases the specificity of DNA binding. In vertebrates, the situation is more complex, with four TCF genes producing many isoforms that contain the HMG domain, but only some of which possess a C-clamp. Vertebrate TCFs have been reported to act in concert with many other transcription factors, which may explain how they obtain sufficient specificity for specific DNA sequences, as well as how they achieve a wide diversity of transcriptional outputs in different cells. PMID- 21624093 TI - The Wnt/Frizzled pathway as a therapeutic target for cardiac hypertrophy: where do we stand? AB - Cardiac hypertrophy is an enlargement of the heart muscle in response to wall stress. This hypertrophic response often leads to heart failure. In recent years, several studies have shown the involvement of Wnt signalling in hypertrophic growth. In this review, the role of Wnt signalling and the possibilities for therapeutic interventions are discussed. In healthy adult heart tissue, Wnt signalling is very low. However, under pathological condition such as hypertension, Wnt signalling is activated. In recent years, it has become clear that both beta-catenin-dependent signalling and beta-catenin-independent signalling are involved in hypertrophic growth. Several studies, both in vitro and in vivo, have shown that genetic interventions in Wnt signalling at different levels resulted in an attenuated or diminished hypertrophic response. Therefore, inhibition of Wnt signalling could provide a new therapeutic strategy for cardiac hypertrophy, but further research on the Wnts and Frizzleds involved in the different forms of cardiac hypertrophy will be needed to achieve this goal. PMID- 21624094 TI - Extracellular nucleotides affect pericyte-mediated regulation of rat in situ vasa recta diameter. AB - AIM: We hypothesized that extracellular nucleotides, established as being released from renal tubular epithelial cells, act at pericytes to regulate vasa recta capillary diameter. METHODS: A rat live kidney slice model and video imaging techniques were used to investigate the effects of extracellular nucleotides on in situ (subsurface) vasa recta diameter at pericyte and non pericyte sites. In addition, RT-qPCR was used to quantify P2 receptor mRNA expression in isolated vasa recta. RESULTS: Extracellular ATP, UTP, benzylbenzyl ATP (BzATP) or 2-methylthioATP (2meSATP) evoked a significantly greater vasoconstriction of subsurface vasa recta at pericytes than at non-pericyte sites. The rank order of agonist potency was BzATP = 2meSATP > ATP = UTP. The vasoconstriction evoked at pericyte sites by ATP was significantly attenuated by the P2 receptor antagonists suramin, pyridoxal phosphate-6-azo(benzene-2,4 disulfonic acid) (PPADS) or Reactive Blue-2 (RB-2). UTP-evoked vasoconstriction at pericytes was attenuated by suramin or RB-2 but not PPADS. Interestingly, suramin or PPADS, when applied in the absence of a P2 receptor agonist, evoked a weak but significant vasoconstriction of vasa recta at pericyte sites, suggesting tonic vasodilation by nucleotides. Significant levels of P2X(1, 3 and 7) and P2Y(4 and 6) receptor mRNA were detected in vasa recta. CONCLUSION: Extracellular nucleotides act at pericytes to cause vasoconstriction of in situ vasa recta. Pharmacological characterization, supported by RT-qPCR data, suggests that P2X(1 and 7) and P2Y(4) receptors mediate nucleotide-evoked vasoconstriction of vasa recta by pericytes. We propose that nucleotides released from renal tubular epithelial cells, in close proximity to vasa recta capillaries, are key in regulating renal medullary blood flow. PMID- 21624096 TI - Writer's cramp: cortical excitability in tasks involving proximo-distal coordination. AB - AIM: The aim of this work was to analyse how writer's cramp patients coordinate each element of the proximal to distal upper arm muscle chain during voluntary movement. METHODS: Using transcranial magnetic stimulation, we have assessed motor cortex excitability properties in patients by recording motor-evoked potentials and silent periods in both the extensor carpi radialis (ECR) and the first dorsal interosseus muscles (FDI), activated either in isolation, or in conjunction with voluntary medial deltoid (MD) co-activation during performance of precise tasks. Ten dystonic patients and ten healthy controls were tested. RESULTS: In both test groups, the ECR muscle displayed a similar active motor threshold, but the excitability curves reached higher plateau values, when the proximal MD muscle was co-activated. In the dystonic group, the FDI muscle excitability curves reached higher plateau values when the MD was co-activated, whereas co-activation had no effect on the control group. In the control group, silent periods, in both the ECR and the FDI were longer when the MD was co activated. This effect was not observed in the dystonic group. CONCLUSION: In the dystonic group, facilitation of the FDI was observed during a task involving proximo-distal coordination. No differences in silent periods were observed when the muscle was activated alone. Our results suggest that such abnormal facilitation is not only an impairment of the central inhibitory mechanisms reported for dystonic patients, but, in addition, represents true abnormality in cortical muscle activation strategies. PMID- 21624095 TI - Integration of transient receptor potential canonical channels with lipids. AB - Transient receptor potential canonical (TRPC) channels are the canonical (C) subset of the TRP proteins, which are widely expressed in mammalian cells. They are thought to be primarily involved in determining calcium and sodium entry and have wide-ranging functions that include regulation of cell proliferation, motility and contraction. The channels are modulated by a multiplicity of factors, putatively existing as integrators in the plasma membrane. This review considers the sensitivities of TRPC channels to lipids that include diacylglycerols, phosphatidylinositol bisphosphate, lysophospholipids, oxidized phospholipids, arachidonic acid and its metabolites, sphingosine-1-phosphate, cholesterol and some steroidal derivatives and other lipid factors such as gangliosides. Promiscuous and selective lipid sensing have been detected. There appear to be close working relationships with lipids of the phospholipase C and A(2) enzyme systems, which may enable integration with receptor signalling and membrane stretch. There are differences in the properties of each TRPC channel that are further complicated by TRPC heteromultimerization. The lipids modulate activity of the channels or insertion in the plasma membrane. Lipid microenvironments and intermediate sensing proteins have been described that include caveolae, G protein signalling, SEC14-like and spectrin-type domains 1 (SESTD1) and podocin. The data suggest that lipid sensing is an important aspect of TRPC channel biology enabling integration with other signalling systems. PMID- 21624097 TI - Inflammatory cytokines in paraventricular nucleus modulate sympathetic activity and cardiac sympathetic afferent reflex in rats. AB - AIM: This study was to determine the roles of inflammatory cytokines in paraventricular nucleus (PVN) in modulating sympathetic activity, blood pressure and cardiac sympathetic afferent reflex (CSAR). METHODS: Renal sympathetic nerve activity (RSNA) and mean arterial pressure (MAP) were recorded in anaesthetized rats with bilateral sinoaortic denervation and vagotomy. The CSAR was evaluated by the RSNA response to epicardial application of bradykinin (BK). The levels of inflammatory cytokines were measured with ELISA. RESULTS: The PVN microinjection of pro-inflammatory cytokines (PIC), tumour necrosis factor (TNF)-alpha or interleukin (IL)-1beta, increased the baseline MAP and RSNA, and enhanced the CSAR. Anti-inflammatory cytokines (AIC), IL-4 or IL-13, in the PVN only increased the baseline MAP. In the rats pretreated with TNF-alpha or IL-1beta but not in the rats pretreated with IL-4 or IL-13, sub-response dose of angiotensin II caused significant increases in the MAP and RSNA and enhancement in the CSAR. AT(1) receptor antagonist losartan in the PVN attenuated the effects of angiotensin II, TNF-alpha and IL-1beta, but not the effects of IL-4 and IL-13. Stimulation of cardiac sympathetic afferents with epicardial application of BK increased the levels of TNF-alpha, IL-1beta but not IL-4 in the PVN. CONCLUSION: TNF-alpha or IL-1beta in the PVN increases blood pressure and sympathetic outflow and enhances the CSAR, which is partially dependent on the AT(1) receptors, while IL-4 or IL-13 in the PVN only increases blood pressure. There is a synergetic effect of Ang II with TNF-alpha or IL-1beta on blood pressure, sympathetic activity and CSAR. PMID- 21624098 TI - Integration of baroreflex and autoregulation control of bronchial blood flow in awake dogs. AB - AIM: Baroreflex control of the bronchial circulation is unresolved. Early studies suggested that baroreflexes dilate or have no effect, but recent studies in awake dogs suggested baroreflexes did not normally engage tonic vasoconstrictor efferents but during excitement systemic pressure rises may also trigger local sensory-motor dilator reflexes. We examined the postulate that bronchial flow is normally regulated at rest during controlled changes in pressure gradient (Pg) by integration of tonic autonomic activity with autoregulation. METHODS: Twelve greyhounds were instrumented under general anaesthesia by surgical implantation of pulsed Doppler flow transducers on the right bronchial artery (BA). After recovery baroreflex effects were evoked by raising and lowering aortic pressure using a lower thoracic aortic balloon in 11 animals, and in six of these after cholinoceptor plus adrenoceptor blockade. RESULTS: The right BA bed showed pressure-passive responses and the time-dependent bronchial bed effects in the autonomically intact state (INT) were largely similar to those in the blocked state (TAB). When results were replotted as pressure-flow relationships and analysed using covariance, the regression line over the pressure range 70-135 mmHg for TAB demonstrated a significant slope (P < 0.05), a linear regression elevated 120% (P = 0.006) above and parallel to INT (Y(Q) = 0.034 + 0.00033(X(Pg) - 104.6). The regression fell on the line of equal proportional change. CONCLUSION: Baroreflexes do not functionally engage the autonomic outflow to the bronchial circulation. Under controlled conditions of systemic Pg change, the bronchial circulation is normally controlled by the integration of resting autonomic tone, myogenic autoregulation and pressure-passive effects. PMID- 21624099 TI - Gas exchange in frogs and turtles: how ectothermic vertebrates contributed to solving the controversy of pulmonary oxygen secretion. AB - The mechanisms governing pulmonary gas exchange were heavily debated at the start of the 20th century when Christian Bohr provided measurements of lung and blood gases as well as rational arguments in favour of oxygen being secreted actively from the lung gas to the blood within vertebrate lungs. The concept of active transport was studied by August Krogh in his doctoral dissertation on the partitioning of gas exchange in frogs. In later studies, where Marie and August Krogh provided conclusive evidence that pulmonary gas exchange occurs by diffusion and diffusion alone, the turtle lungs provided an important tool to investigate the role of perfusion in pulmonary gas exchange. Here, I review the early Bohr and Krogh studies on pulmonary and cutaneous gas exchange in frogs as well as the experimental studies on gas exchange and its possible autonomic regulation in turtles. The results are discussed within the context of recent studies on the cardiorespiratory physiology of frogs and turtles. PMID- 21624100 TI - Effects of adrenaline on lactate, glucose, lipid and protein metabolism in the placebo controlled bilaterally perfused human leg. AB - AIM: Adrenaline has widespread metabolic actions, including stimulation of lipolysis and induction of insulin resistance and hyperlactatemia. Systemic adrenaline administration, however, generates a very complex hormonal and metabolic scenario. No studies employing regional, placebo controlled and adrenaline infusion exist. Our study was designed to test the hypothesis that local placebo controlled leg perfusion with adrenaline directly increases local lactate release, stimulates lipolysis, induces insulin resistance and leaves protein metabolism unaffected. METHODS: We studied seven healthy volunteers with bilateral femoral vein and artery catheters during 3-h basal and 3-h hyperinsulinemic (0.6 mU kg(-1) min(-1) ) euglycemic clamp conditions. One femoral artery was perfused with saline and the other with adrenaline (0.4 MUg min m(-2) ). Lipid metabolism was quantified with [9,10-(3) H] palmitate and amino acid metabolism with (15) N-phenylalanine and lactate and glucose by raw arterio-venous differences. RESULTS: Femoral vein plasma adrenaline increased ~eightfold in the perfused leg with unaltered blood flows. Adrenaline perfusion significantly increased local leg lactate release from 0.01 to 0.25 mmol min(-1) per leg, palmitate release in the basal state 11.5-16.9 MUmol min(-1) per leg and during the clamp 2.62-8.44 MUmol min(-1) per leg. Glucose uptake decreased during the clamp from ~180 to 30 MUmol min(-1) per leg. Phenylalanine kinetics was not affected by adrenaline. CONCLUSION: Adrenaline directly increases lactate release and lipolysis and inhibits insulin-stimulated glucose uptake in the perfused human leg. Adrenaline has no direct effects on peripheral amino acid metabolism. Adrenaline-induced lactate release from striated muscle may be an important mechanism underlying hyperlactatemia in the critically ill. PMID- 21624101 TI - Health-related quality of life scores in long-term head and neck cancer survivors predict subsequent survival: a prospective cohort study. AB - OBJECTIVE: To examine the survival prediction of long-term health-related quality of life in patients with head and neck squamous cell carcinoma. DESIGN: Patients with head and neck squamous cell carcinoma diagnosed in the period between July 1992 and October 2001, who had been disease free for a minimum of 1 year following therapy, responded to structured interviews including several validated questionnaires in the period from October 2002 to March 2004. The study ended in June 2009 with a mean observation time of 75 +/- 4 months among the survivors. Twenty-four deaths were observed. SETTING: University hospital, referral centre of the Western Norway. PARTICIPANTS: One hundred and thirty-nine cognitive functioning patients. MAIN OUTCOME MEASUREMENTS: Overall survival as of June 2009. This was correlated with various clinical factors and the EORTC QLQ-C30 questionnaire, the Eysenck Personality Inventory and the Coping inventory completed between October 2001 and March 2004. RESULTS: A general symptom sum score was significantly predictive of survival directly and after sequential adjustment for self-reported levels of neuroticism, avoidance focused coping, coping by suppression of competing activity, alcohol consumption, smoking status and heart/lung disease, as well as gender, age, time between diagnosis and inclusion, tumour node metastasis (TNM) stage and tumour site. Similar results were found for the health-related quality of life indices 'fatigue', 'dyspnoea' and 'sleep disturbance'. A dichotomised variable based on the general symptom sum score was calculated, and a high risk group, as to mortality, including less than a quintile of the total patient population was established. A hazard ratio of 5.15 was found for the dichotomised general symptom sum score. CONCLUSION: We have shown a unique and independent survival prediction from long-term EORTC QLQ C30 scores in successfully treated and cognitive functioning head and neck squamous cell carcinoma patients. PMID- 21624102 TI - First evidence of mineralization of petroleum asphaltenes by a strain of Neosartorya fischeri. AB - A fungal strain isolated from a microbial consortium growing in a natural asphalt lake is able to grow in purified asphaltenes as the only source of carbon and energy. The asphaltenes were rigorously purified in order to avoid contamination from other petroleum fractions. In addition, most of petroporphyrins were removed. The 18S rRNA and beta-tubulin genomic sequences, as well as some morphologic characteristics, indicate that the isolate is Neosartorya fischeri. After 11 weeks of growth, the fungus is able to metabolize 15.5% of the asphaltenic carbon, including 13.2% transformed to CO(2) . In a medium containing asphaltenes as the sole source of carbon and energy, the fungal isolate produces extracellular laccase activity, which is not detected when the fungus grow in a rich medium. The results obtained in this work clearly demonstrate that there are microorganisms able to metabolize and mineralize asphaltenes, which is considered the most recalcitrant petroleum fraction. PMID- 21624103 TI - A novel screening system for secretion of heterologous proteins in Bacillus subtilis. AB - High-level production of secretory proteins in Bacillus subtilis leads to a stress response involving the two-component system CssRS and its target genes htrA and htrB. Here, we used this sensing system in a reporter strain in which gfp is under control of P(htrA) , the secretion stress responsive promoter of htrA. Overexpression of heterologous secretory proteins in this strain results in green fluorescent cells, which can be separated from non-secreting, low fluorescent cells using a fluorescence-activated cell sorter (FACS). Using this principle, genomic libraries of uncharacterized prokaryotic organisms, expressed in the reporter strain, can be screened for genes encoding secretory proteins. PMID- 21624104 TI - Optimal target refraction for implantation of monofocal intraocular lenses. PMID- 21624105 TI - White spot syndrome virus (WSSV) concentrations in crustacean tissues: a review of data relevant to assess the risk associated with commodity trade. AB - We have reviewed the available peer reviewed literature on pathogen load for white spot syndrome virus (WSSV) in species susceptible to infection. Data on pathogen load in traded commodities are relevant for undertaking import risk assessments for a specific pathogen. Data were available for several of the major penaeid shrimp species farmed for aquaculture and for one crab and crayfish species. Most data are based on experimental infection, but some data were available for farmed or wild shrimp. Owing to the unavailability of immortal cell lines to determine viral load of viable virus, quantitative PCR was the main method used for quantification. The viral loads measured in shrimp at the onset of mortality events were extremely high (in the order of 10(9) -10(10) copy numbers gram(-1) of tissue). In a farm setting, the onset of increased mortalities will often trigger emergency harvests. Therefore, shrimp obtained from emergency harvests are likely to carry substantial concentrations of viral particles. Viral load did not vary greatly with tissue type. The WSSV load in wild crustaceans, farmed crustaceans not undergoing a mortality event or survivors of a mortality event was significantly lower (usually by multiple logs). Studies have also been undertaken in 'vaccinated' shrimp. One of the 'vaccines' led to a significant reduction of viral load in WSSV-exposed animals. The data obtained from the literature review are put into context with published information on minimal infectious dose and WSSV survival in frozen commodity shrimp. PMID- 21624106 TI - On-farm study of human contact networks to document potential pathways for avian influenza transmission between commercial poultry farms in Ontario, Canada. AB - Human movements associated with poultry farming create contact networks that might facilitate transmission of avian influenza (AI) between farms during outbreaks. In Canada, no information is available about how these networks connect poultry farms. The purpose of this study was to document human contacts between commercial poultry farms in Ontario, Canada, to learn how AI might be transmitted during outbreaks. We used face-to-face interviews with people entering the farm biosecurity perimeter on four layer, one turkey and three broiler breeder poultry farms in Ontario to collect information on between-farm contacts and biosecurity practices. Over a four-day study period on each farm, a median of 10.5 people entered the farm biosecurity perimeter (range 2-31). Ninety six per cent (111/118) of people consented to be interviewed. Of these, fifty three per cent (59/111) had contact with one or more (median 2, degree range 1 14) other poultry farms within 72 h. A median of 25 (range 7-65) human contacts linked study farms to other poultry farms. The mean distance of between-farm contacts was 53 km. Eighty-six per cent of people who answered the biosecurity questions (94/109) reported using one or more biosecurity practices. However, on 7/8 farms, at least one person reported that they did not use any biosecurity practices. Fifty per cent of social visitors used biosecurity, whereas 96% of all other people used biosecurity. Ninety-two per cent of people that entered the poultry barns (46/50) used one or more biosecurity practices, whereas 81% of people (48/59) that did not enter the poultry barns used one or more biosecurity practices. Because our study documented farm visitors who did not use any biosecurity practices and moved between commercial poultry farms, we suggest that rapid trace-out of human movements is as important as containment zoning to limiting disease spread during an outbreak of highly pathogenic AI in Ontario. PMID- 21624107 TI - Measurement of gastric accommodation: a reappraisal of conventional and emerging modalities. AB - Impaired gastric accommodation is an important cause of functional dyspepsia. Currently available tests that evaluate gastric accommodation provide relevant physiological information, but they pose technical difficulties and their clinical impact remains controversial. Gastric barostat remains the gold standard, but it is an invasive procedure. In recent years, emerging modalities including single photon emission computed tomography (SPECT), three-dimensional ultrasound and magnetic resonance imaging have been developed to measure gastric volumes and hold promise as alternative methods of assessing gastric accommodation non-invasively. Studies are underway to validate these techniques with recent data proving the performance characteristics of SPECT. The non invasive nutrient drink test measures satiety scores as a surrogate marker of gastric accommodation and remains controversial. More recently, intragastric monitoring has been proposed as yet another non-invasive modality to assess gastric accommodation. Each of these different modalities brings its associated advantages and disadvantages, as is discussed in this review. Ongoing studies to validate these new techniques are in progress and are likely to lead to further progress in neurogastroenterology. PMID- 21624108 TI - Predicting outcome of rethoracotomy for suspected pericardial tamponade following cardio-thoracic surgery in the intensive care unit. AB - OBJECTIVES: Pericardial tamponade after cardiac surgery is difficult to diagnose, thereby rendering timing of rethoracotomy hard. We aimed at identifying factors predicting the outcome of surgery for suspected tamponade after cardio-thoracic surgery, in the intensive care unit (ICU). METHODS: Twenty-one consecutive patients undergoing rethoracotomy for suspected pericardial tamponade in the ICU, admitted after primary cardio-thoracic surgery, were identified for this retrospective study. We compared patients with or without a decrease in severe haemodynamic compromise after rethoracotomy, according to the cardiovascular component of the sequential organ failure assessment (SOFA) score. RESULTS: A favourable haemodynamic response to rethoracotomy was observed in 11 (52%) of patients and characterized by an increase in cardiac output, and less fluid and norepinephrine requirements. Prior to surgery, the absence of treatment by heparin, a minimum cardiac index < 1.0 L/min/m2 and a positive fluid balance (> 4,683 mL) were predictive of a beneficial haemodynamic response. During surgery, the evacuation of clots and > 500 mL of pericardial fluid was associated with a beneficial haemodynamic response. Echocardiographic parameters were of limited help in predicting the postoperative course, even though 9 of 13 pericardial clots found at surgery were detected preoperatively. CONCLUSION: Clots and fluids in the pericardial space causing regional tamponade and responding to surgical evacuation after primary cardio-thoracic surgery, are difficult to diagnose preoperatively, by clinical, haemodynamic and even echocardiographic evaluation in the ICU. Only absence of heparin treatment, a large positive fluid balance and low cardiac index predicted a favourable haemodynamic response to rethoracotomy. These data might help in deciding and timing of reinterventions after primary cardio-thoracic surgery. PMID- 21624109 TI - Improvement of platelet dysfunction in chronic myelogenous leukemia following treatment with imatinib: a case report. AB - INTRODUCTION: In patients with chronic myeloid leukemia, tyrosine kinase inhibitors suppress the BCR-ABL+ clone and often induce complete molecular remissions. Megakaryocytes in such patients have been shown to be derived from the BCR-ABL+ clone, and abnormal platelet function is frequent in chronic myeloid leukemia. However, little is known about the influence of modern targeted therapy on chronic myeloid leukemia-associated platelet disorders. CASE PRESENTATION: We report the case of a massive hemorrhage in a 32-year-old Caucasian man caused by chronic myeloid leukemia-associated platelet dysfunction, which improved after treatment with imatinib. CONCLUSION: This report demonstrates that platelet dysfunction and bleeding disorder in BCR-ABL+ chronic myeloid leukemia can successfully be treated with imatinib. We suggest the monitoring of platelet function in future studies using imatinib to treat patients with chronic myeloid leukemia. PMID- 21624110 TI - MDM2 antagonist Nutlin-3a potentiates antitumour activity of cytotoxic drugs in sarcoma cell lines. AB - BACKGROUND: Frequent failure and severe side effects of current sarcoma therapy warrants new therapeutic approaches. The small-molecule MDM2 antagonist Nutlin-3a activates the p53 pathway and efficiently induces apoptosis in tumours with amplified MDM2 gene and overexpression of MDM2 protein. However, the majority of human sarcomas have normal level of MDM2 and the therapeutic potential of MDM2 antagonists in this group is still unclear. We have investigated if Nutlin-3a could be employed to augment the response to traditional therapy and/or reduce the genotoxic burden of chemotherapy. METHODS: A panel of sarcoma cell lines with different TP53 and MDM2 status were treated with Nutlin-3a combined with Doxorubicin, Methotrexate or Cisplatin, and their combination index determined. RESULTS: Clear synergism was observed when Doxorubicin and Nutlin-3a were combined in cell lines with wild-type TP53 and amplified MDM2, or with Methotrexate in both MDM2 normal and amplified sarcoma cell lines, allowing for up to tenfold reduction of cytotoxic drug dose. Interestingly, Nutlin-3a seemed to potentiate the effect of classical drugs as Doxorubicin and Cisplatin in cell lines with mutated TP53, but inhibited the effect of Methotrexate. CONCLUSION: The use of Nutlin in combination with classical sarcoma chemotherapy shows promising preclinical potential, but since clear biomarkers are still lacking, clinical trials should be followed up with detailed tumour profiling. PMID- 21624111 TI - Clinical and cognitive correlates of employment among patients with schizophrenia: a cross-sectional study in Malaysia. AB - BACKGROUND: Gainful employment is one major area of functioning which is becoming an important goal in psychiatric rehabilitation of patients with schizophrenia. Studies in western countries are pointing to evidence that certain sociodemographic and clinical factors may contribute to employment outcomes in this group of people. However, the area is still largely unexplored in Malaysia. The aim of this study was to examine the sociodemographic, clinical and cognitive correlates of employment status among patients with Schizophrenia. METHODS: This was a cross-sectional study. All participants who fulfilled the requirements of the study according to the inclusion and exclusion criteria were enrolled. Study instruments included a demographic data questionnaire, Positive and Negative Symptom Scale (PANSS), Trail Making Tests, Rey's Auditory Verbal Learning Test (RAVLT) and Digit Span. Bivariate analyses were done using chi-square for categorical data and t-test for continuous data and multiple logistic regression analysis was done to identify predictors of employment status. RESULTS: A total of 95 participants who fulfilled the inclusion criteria were enrolled into the study. Among the sociodemographic, clinical and cognitive variables studied marital status, educational level, mean scores of negative symptoms, Digit Span and RAVLT and Trail Making Tests were found to show significant association with employment status on bivariate analyses. However, when entered into a logistic regression model, only cognitive variables ie. Trail A and B, Digit Span and RAVLT were significant predictors of employment status. CONCLUSIONS: The results from this study support the role of cognitive function, particularly, attention, working memory and executive functioning on attaining and maintaining employment in persons with schizophrenia as measured by the RAVLT, Digit Span and Trail Making Tests. These findings may act as preliminary evidence suggesting the importance of integrating cognitive rehabilitation in the psychosocial rehabilitation program for patients with schizophrenia in Malaysia. PMID- 21624112 TI - Most bowel cancer symptoms do not indicate colorectal cancer and polyps: a systematic review. AB - BACKGROUND: Bowel symptoms are often considered an indication to perform colonoscopy to identify or rule out colorectal cancer or precancerous polyps. Investigation of bowel symptoms for this purpose is recommended by numerous clinical guidelines. However, the evidence for this practice is unclear. The objective of this study is to systematically review the evidence about the association between bowel symptoms and colorectal cancer or polyps. METHODS: We searched the literature extensively up to December 2008, using MEDLINE and EMBASE and following references. For inclusion in the review, papers from cross sectional, case control and cohort studies had to provide a 2*2 table of symptoms by diagnosis (colorectal cancer or polyps) or sufficient data from which that table could be constructed. The search procedure, quality appraisal, and data extraction was done twice, with disagreements resolved with another reviewer. Summary ROC analysis was used to assess the diagnostic performance of symptoms to detect colorectal cancer and polyps. RESULTS: Colorectal cancer was associated with rectal bleeding (AUC 0.66; LR+ 1.9; LR- 0.7) and weight loss (AUC 0.67, LR+ 2.5, LR- 0.9). Neither of these symptoms was associated with the presence of polyps. There was no significant association of colorectal cancer or polyps with change in bowel habit, constipation, diarrhoea or abdominal pain. Neither the clinical setting (primary or specialist care) nor study type was associated with accuracy.Most studies had methodological flaws. There was no consistency in the way symptoms were elicited or interpreted in the studies. CONCLUSIONS: Current evidence suggests that the common practice of performing colonoscopies to identify cancers in people with bowel symptoms is warranted only for rectal bleeding and the general symptom of weight loss. Bodies preparing guidelines for clinicians and consumers to improve early detection of colorectal cancer need to take into account the limited value of symptoms. PMID- 21624113 TI - Co-expression of ubiquitin gene and capsid protein gene enhances the potency of DNA immunization of PCV2 in mice. AB - A recombinant plasmid that co-expressed ubiquitin and porcine circovirus type 2 (PCV2) virus capsid protein (Cap), denoted as pc-Ub-Cap, and a plasmid encoding PCV2 virus Cap alone, denoted as pc-Cap, were transfected into 293T cells. Indirect immunofluorescence (IIF) and confocal microscopy were performed to measure the cellular expression of Cap. Three groups of mice were then vaccinated once every three weeks for a total of three doses with pc-Ub-Cap, pc-Cap or the empty vector pCAGGS, followed by challenging all mice intraperitoneally with 0.5 mL 106.5 TCID50/mL PCV2. To characterize the protective immune response against PCV2 infection in mice, assays of antibody titer (including different IgG isotypes), flow cytometric analysis (FCM), lymphocyte proliferation, cytokine production and viremia were evaluated. The results showed that pc-Ub-Cap and pc Cap were efficiently expressed in 293T cells. However, pc-Ub-Cap-vaccinated animals had a significantly higher level of Cap-specific antibody and induced a stronger Th1 type cellular immune response than did pc-Cap-vaccinated animals, suggesting that ubiquitin conjugation improved both the cellular and humoral immune responses. Additionally, viral replication in blood was lower in the pc-Ub Cap-vaccinated group than in the pc-Cap and empty vector groups, suggesting that the protective immunity induced by pc-Ub-Cap is superior to that induced by pc Cap. PMID- 21624114 TI - Study of polymorphisms in tir, eae and tccP2 genes in enterohaemorrhagic and enteropathogenic Escherichia coli of serogroup O26. AB - BACKGROUND: Enteropathogenic (EPEC) and enterohaemorrhagic (EHEC) Escherichia coli are responsible for food poisoning (enteritis and enterotoxaemia) in humans in developed countries. Cattle are considered to be an important reservoir of EHEC and EPEC strains for humans. Moreover, some of the strains, belonging to the O26, O111, O118 serogroups, for example, are also responsible for digestive disorders in calves. The Translocated intimin receptor (Tir), the intimin (Eae) and the Tir-cytoskeleton coupling protein (TccP) represent three virulence factors implicated in the intimate attachment of the bacteria to the eukaryotic cell. Major variants have already been described for these genes among the different serogroups but minor variations have not often been studied. In this study, we examined the polymorphisms of the tir, eae and tccP2 genes of O26 strains (EPEC and EHEC isolated from bovines and from humans) with the aim to determine whether these polymorphisms are host specific or not. RESULTS: Of the 70 tested strains, 10 strains (14% of the strains) presented one or several polymorphisms in the tir and eae genes, which have never previously been described. Concerning tccP2 detection, 47 of the 70 strains (67% of the strains) were found to be positive for this gene. Most of the strains were found to possess tccP2 variants described in strains of serogroup O26. Nevertheless, three strains had tccP2 genes respectively described in strains of serogroup O111, O103 and O55. Moreover, none of the polymorphisms was statistically specific to the bovine or the human isolates. Nevertheless, the two major variants of tccP2 were statistically associated with the pathotype (EPEC or EHEC). CONCLUSIONS: In conclusion, tir and eae gene polymorphisms were found not to be numerous and not to be predominantly synonymous. Moreover, no difference was observed between human and bovine strains regarding the presence of polymorphisms. Finally, some tccP2 variants appeared to be pathotype specific. Further investigations need to be performed on a larger number of strains in order to confirm this specificity. PMID- 21624115 TI - Systematic quantitative characterization of cellular responses induced by multiple signals. AB - BACKGROUND: Cells constantly sense many internal and environmental signals and respond through their complex signaling network, leading to particular biological outcomes. However, a systematic characterization and optimization of multi-signal responses remains a pressing challenge to traditional experimental approaches due to the arising complexity associated with the increasing number of signals and their intensities. RESULTS: We established and validated a data-driven mathematical approach to systematically characterize signal-response relationships. Our results demonstrate how mathematical learning algorithms can enable systematic characterization of multi-signal induced biological activities. The proposed approach enables identification of input combinations that can result in desired biological responses. In retrospect, the results show that, unlike a single drug, a properly chosen combination of drugs can lead to a significant difference in the responses of different cell types, increasing the differential targeting of certain combinations. The successful validation of identified combinations demonstrates the power of this approach. Moreover, the approach enables examining the efficacy of all lower order mixtures of the tested signals. The approach also enables identification of system-level signaling interactions between the applied signals. Many of the signaling interactions identified were consistent with the literature, and other unknown interactions emerged. CONCLUSIONS: This approach can facilitate development of systems biology and optimal drug combination therapies for cancer and other diseases and for understanding key interactions within the cellular network upon treatment with multiple signals. PMID- 21624116 TI - The synthetic peptide P111-136 derived from the C-terminal domain of heparin affin regulatory peptide inhibits tumour growth of prostate cancer PC-3 cells. AB - BACKGROUND: Heparin affin regulatory peptide (HARP), also called pleiotrophin, is a heparin-binding, secreted factor that is overexpressed in several tumours and associated to tumour growth, angiogenesis and metastasis. The C-terminus part of HARP composed of amino acids 111 to 136 is particularly involved in its biological activities and we previously established that a synthetic peptide composed of the same amino acids (P111-136) was capable of inhibiting the biological activities of HARP. Here we evaluate the ability of P111-136 to inhibit in vitro and in vivo the growth of a human tumour cell line PC-3 which possess an HARP autocrine loop. METHODS: A total lysate of PC-3 cells was incubated with biotinylated P111-136 and pulled down for the presence of the HARP receptors in Western blot. In vitro, the P111-136 effect on HARP autocrine loop in PC-3 cells was determined by colony formation in soft agar. In vivo, PC-3 cells were inoculated in the flank of athymic nude mice. Animals were treated with P111-136 (5 mg/kg/day) for 25 days. Tumour volume was evaluated during the treatment. After the animal sacrifice, the tumour apoptosis and associated angiogenesis were evaluated by immunohistochemistry. In vivo anti-angiogenic effect was confirmed using a mouse MatrigelTM plug assay. RESULTS: Using pull down experiments, we identified the HARP receptors RPTPbeta/zeta, ALK and nucleolin as P111-136 binding proteins. In vitro, P111-136 inhibits dose dependently PC-3 cell colony formation. Treatment with P111-136 inhibits significantly the PC-3 tumour growth in the xenograft model as well as tumour angiogenesis. The angiostatic effect of P111-136 on HARP was also confirmed using an in vivo MatrigelTM plug assay in mice CONCLUSIONS: Our results demonstrate that P111-136 strongly inhibits the mitogenic effect of HARP on in vitro and in vivo growth of PC-3 cells. This inhibition could be linked to a direct or indirect binding of this peptide to the HARP receptors (ALK, RPTPbeta/zeta, nucleolin). In vivo, the P111-136 treatment significantly inhibits both the PC-3 tumour growth and the associated angiogenesis. Thus, P111-136 may be considered as an interesting pharmacological tool to interfere with tumour growth that has now to be evaluated in other cancer types. PMID- 21624117 TI - Community perceptions of malaria and vaccines in the South Coast and Busia regions of Kenya. AB - BACKGROUND: Malaria is a leading cause of morbidity and mortality in children younger than 5 years in Kenya. Within the context of planning for a vaccine to be used alongside existing malaria control methods, this study explores sociocultural and health communications issues among individuals who are responsible for or influence decisions on childhood vaccination at the community level. METHODS: This qualitative study was conducted in two malaria-endemic regions of Kenya--South Coast and Busia. Participant selection was purposive and criterion based. A total of 20 focus group discussions, 22 in-depth interviews, and 18 exit interviews were conducted. RESULTS: Participants understand that malaria is a serious problem that no single tool can defeat. Communities would welcome a malaria vaccine, although they would have questions and concerns about the intervention. While support for local child immunization programs exists, limited understanding about vaccines and what they do is evident among younger and older people, particularly men. Even as health care providers are frustrated when parents do not have their children vaccinated, some parents have concerns about access to and the quality of vaccination services. Some women, including older mothers and those less economically privileged, see themselves as the focus of health workers' negative comments associated with either their parenting choices or their children's appearance. In general, parents and caregivers weigh several factors--such as personal opportunity costs, resource constraints, and perceived benefits--when deciding whether or not to have their children vaccinated, and the decision often is influenced by a network of people, including community leaders and health workers. CONCLUSIONS: The study raises issues that should inform a communications strategy and guide policy decisions within Kenya on eventual malaria vaccine introduction. Unlike the current practice, where health education on child welfare and immunization focuses on women, the communications strategy should equally target men and women in ways that are appropriate for each gender. It should involve influential community members and provide needed information and reassurances about immunization. Efforts also should be made to address concerns about the quality of immunization services--including health workers' interpersonal communication skills. PMID- 21624118 TI - Health related quality of life among Iraqi immigrants settled in Malaysia. AB - BACKGROUND: Migrants everywhere face several demands for health and maintaining good health and quality of life could be challenging. Iraqis are the second largest refugee group that has sought refuge in the recent years, yet little is known about their health related quality of life (HRQOL). The study aims at assessing the HRQOL among Iraqis living in Malaysia. METHODS: A self-administered Arabic version of Sf-36 questionnaire was distributed among 300 Iraqi migrants in Malaysia. The questionnaire taps eight concepts of physical and mental health to assess the HRQOL. Univariate analysis was performed for group analysis (t test, ANOVA) and Multiple Linear Regression was used to control for confounding effects. RESULTS: Two hundred and fifty three participants ranging in age from 18 to 67 years (Mean = 33.6) returned the completed questionnaire. The majority was males (60.1%) and more than half of the respondents (59.5%) were married. Less than half (45.4%) and about a quarter (25.9%) reported bachelor degree and secondary school education respectively and the remaining 28.7% had either a master or a PhD degree.Univariate analysis showed that the HRQOL scores among male immigrants were found to be higher than those of females in physical function (80.0 vs. 73.5), general health (72.5 vs. 60.7) and bodily pain (87.9 vs. 72.5) subscales. The youngest age group had significantly higher physical function (79.32) and lower mental health scores (57.62).The mean score of physical component summary was higher than the mental component summary mean score (70.22 vs. 63.34).Stepwise multiple linear regression, revealed that gender was significantly associated with physical component summary (beta = - 6.06, p = 0.007) and marital status was associated with mental component summary (beta = 7.08, p = 0.003). CONCLUSIONS: From the data it appears that Iraqi immigrants living in Malaysia have HRQOL scores that might be considered to indicate a relatively moderate HRQOL. The HRQOL is significantly affected by gender and marital status. Further studies are needed to explore determinants of HRQOL consequent to immigration. The findings could be worthy of further exploration. PMID- 21624119 TI - Effect of clone selection, nitrogen supply, leaf damage and mycorrhizal fungi on stilbene and emodin production in knotweed. AB - BACKGROUND: Fallopia japonica and its hybrid, F. xbohemica, due to their fast spread, are famous as nature threats rather than blessings. Their fast growth rate, height, coverage, efficient nutrient translocation between tillers and organs and high phenolic production, may be perceived either as dangerous or beneficial features that bring about the elimination of native species or a life supporting source. To the best of our knowledge, there have not been any studies aimed at increasing the targeted production of medically desired compounds by these remarkable plants. We designed a two-year pot experiment to determine the extent to which stilbene (resveratrol, piceatannol, resveratrolosid, piceid and astringins) and emodin contents of F. japonica, F. sachalinensis and two selected F. xbohemica clones are affected by soil nitrogen (N) supply, leaf damage and mycorrhizal inoculation. RESULTS: 1) Knotweeds are able to grow on substrates with extremely low nitrogen content and have a high efficiency of N translocation. The fast-spreading hybrid clones store less N in their rhizomes than the parental species. 2) The highest concentrations of stilbenes were found in the belowground biomass of F. japonica. However, because of the high belowground biomass of one clone of F. xbohemica, this hybrid produced more stilbenes per plant than F. japonica. 3) Leaf damage increased the resveratrol and emodin contents in the belowground biomass of the non-inoculated knotweed plants. 4) Although knotweed is supposed to be a non-mycorrhizal species, its roots are able to host the fungi. Inoculation with mycorrhizal fungi resulted in up to 2% root colonisation. 5) Both leaf damage and inoculation with mycorrhizal fungi elicited an increase of the piceid (resveratrol-glucoside) content in the belowground biomass of F. japonica. However, the mycorrhizal fungi only elicited this response in the absence of leaf damage. Because the leaf damage suppressed the effect of the root fungi, the effect of leaf damage prevailed over the effect of the mycorrhizal fungi on the piceid content in the belowground biomass. CONCLUSIONS: Two widely spread knotweed species, F. japonica and F. xbohemica, are promising sources of compounds that may have a positive impact on human health. The content of some of the target compounds in the plant tissues can be significantly altered by the cultivation conditions including stress imposed on the plants, inoculation with mycorrhizal fungi and selection of the appropriate plant clone. PMID- 21624120 TI - Relationship between time-integrated disease activity estimated by DAS28-CRP and radiographic progression of anatomical damage in patients with early rheumatoid arthritis. AB - BACKGROUND: The main aim of the study was to investigate the relationship between persistent disease activity and radiographic progression of joint damage in early rheumatoid arthritis (ERA). METHODS: Forty-eight patients with active ERA was assessed every 3 months for disease activity for 3 years. Radiographic damage was measured by the Sharp/van der Heijde method (SHS). The cumulative inflammatory burden was estimated by the time-integrated values (area under the curve-AUC) of Disease Activity Score 28 joint based on C-reactive protein (DAS28-CRP) in rapid progressors versus non-progressors. Bland and Altman's 95% limits of agreement method were used to estimate the smallest detectable difference (SDD) of radiographic progression. The relationship between clinical and laboratory predictors of radiographic progression and their interactions with time was analysed by logistic regression model. RESULTS: After 3-years of follow-up, radiographic progression was observed in 54.2% (95%CI: 39.8% to 67.5%) of patients and SDD was 9.5 for total SHS. The percentage of patients with erosive disease increased from 33.3% at baseline to 76% at 36 months. The total SHS of the progressors worsened from a median (interquartile range) of 18.5 (15-20) at baseline to 38.5 (34-42) after 3 years (p < 0.0001) whereas non-progressors worsened from a median of 14.5 (13-20) at baseline to 22.5 (20-30) after 3 years (p < 0.001). In the regression model, time-integrated values of DAS28-CRP and anti-CCP positivity have the highest positive predictive value for progression (both at level of p < 0.0001). Radiographic progression was also predicted by a positive IgM-RF (p0.0009), and a high baseline joint damage (p = 0.0044). CONCLUSIONS: These data indicate that the level of disease activity, as measured by time-integrated DAS28-CRP, anti-CCP and IgM-RF positivity and a high baseline joint damage, affects subsequent progression of radiographic damage in ERA. PMID- 21624121 TI - Prognostic value of the expression of C-Chemokine Receptor 6 and 7 and their ligands in non-metastatic breast cancer. AB - BACKGROUND: Chemokines and chemokine receptors are major actors of leukocytes trafficking and some have been shown to play an important role in cancer metastasis. Chemokines CCL19, CCL20 and CCL21 and their receptors CCR6 and CCR7, were assessed as potential biomarkers of metastatic dissemination in primary breast cancer. METHODS: Biomarker expression levels were evaluated using immunohistochemistry on paraffin-embedded tissue sections of breast cancer (n = 207). RESULTS: CCR6 was expressed by tumor cells in 35% of cases. CCR7 was expressed by spindle shaped stromal cells in 43% of cases but not by tumor cells in this series. CCL19 was the only chemokine found expressed in a significant number of breast cancers and was expressed by both tumor cells and dendritic cells (DC). CCR6, CCL19 and CCR7 expression correlated with histologic features of aggressive disease. CCR6 expression was associated with shorter relapse-free survival (RFS) in univariate and but not in multivariate analysis (p = 0.0316 and 0.055 respectively), and was not associated with shorter overall survival (OS). Expression of CCR7 was not significantly associated with shorter RFS or OS. The presence of CCL19-expressing DC was associated with shorter RFS in univariate and multivariate analysis (p = 0.042 and 0.020 respectively) but not with shorter OS. CONCLUSION: These results suggest a contribution of CCR6 expression on tumor cells and CCL19-expressing DC in breast cancer dissemination. In our series, unlike what was previously published, CCR7 was exclusively expressed on stromal cells and was not associated with survival. PMID- 21624122 TI - Ethnic variation in validity of the estimated obesity prevalence using self reported weight and height measurements. AB - BACKGROUND: We examined ethnic differences between levels of body mass index (BMI) based on self-reported and measured body height and weight and the validity of self-reports used to estimate the prevalence of obesity (BMI>=30 kg/m2) in Turkish, Moroccan, and Dutch people in the Netherlands. Furthermore, we investigated whether BMI levels and the prevalence of obesity in Turkish and Moroccan people with incomplete self-reports (missing height or weight) differ from those with complete self-reports. METHODS: Data on self-reported and measured height and weight were collected in a population-based survey among 441 Dutch, 414 Turks and 344 Moroccans aged 18 to 69 years in Amsterdam, the Netherlands in 2004. BMI and obesity were calculated from self-reported and measured height and weight. RESULTS: The difference between measured and estimated BMI was larger in Turkish and Moroccan women than in Dutch women, which was explained by the higher BMI of the Turkish and Moroccan women. In men we found no ethnic differences between measured and estimated BMI. Sensitivity to detect obesity was low and specificity was high. In participants with available self-reported and measured height and weight, self-reports produced a similar underestimation of the obesity prevalence in all ethnic groups. However, many obese Turkish and Moroccan women had incomplete self-reports, missing height or weight, resulting in an additional underestimation of the prevalence of obesity. Among men (all ethnicities) and Dutch women, the availability of height or weight by self-report did not differ between obese and non obese participants. CONCLUSIONS: BMI based on self-reports is underestimated more by Turkish and Moroccan women than Dutch women, which is explained by the higher BMI of Turkish and Moroccan women. Further, in women, ethnic differences in the estimation of obesity prevalence based on self-reports do exist and are due to incomplete self reports in obese Turkish and Moroccan women. In men, ethnicity is not associated with discrepancies between levels of BMI and obesity prevalence based on measurements and self-reports. Hence, our results indicate that using measurements to accurately determine levels of BMI and obesity prevalence in public health research seems even more important in Turkish and Moroccan migrant women than in other populations. PMID- 21624123 TI - Differential roles of Smad2 and Smad3 in the regulation of TGF-beta1-mediated growth inhibition and cell migration in pancreatic ductal adenocarcinoma cells: control by Rac1. AB - BACKGROUND: Progression of pancreatic ductal adenocarcinoma (PDAC) is largely the result of genetic and/or epigenetic alterations in the transforming growth factor beta (TGF-beta)/Smad signalling pathway, eventually resulting in loss of TGF-beta mediated growth arrest and an increase in cellular migration, invasion, and metastasis. These cellular responses to TGF-beta are mediated solely or partially through the canonical Smad signalling pathway which commences with activation of receptor-regulated Smads (R-Smads) Smad2 and Smad3 by the TGF-beta type I receptor. However, little is known on the relative contribution of each R-Smad, the possible existence of functional antagonism, or the crosstalk with other signalling pathways in the control of TGF-beta1-induced growth inhibition and cell migration. Using genetic and pharmacologic approaches we have inhibited in PDAC cells endogenous Smad2 and Smad3, as well as a potential regulator, the small GTPase Rac1, and have analysed the consequences for TGF-beta1-mediated growth inhibition and cell migration (chemokinesis). RESULTS: SiRNA-mediated silencing of Smad3 in the TGF-beta responsive PDAC cell line PANC-1 reduced TGF beta1-induced growth inhibition but increased the migratory response, while silencing of Smad2 enhanced growth inhibition but decreased chemokinesis. Interestingly, siRNA-mediated silencing of the small GTPase Rac1, or ectopic expression of a dominant-negative Rac1 mutant largely mimicked the effect of Smad2 silencing on both TGF-beta1-induced growth inhibition, via upregulation of the cdk inhibitor p21WAF1, and cell migration. Inhibition of Rac1 activation reduced both TGF-beta1-induction of a Smad2-specific transcriptional reporter and Smad2 C-terminal phosphorylation in PDAC cells while Smad3-specific transcriptional activity and Smad3 C-terminal phosphorylation appeared increased. Disruption of autocrine TGF-beta signalling in PANC-1 cells rendered cells less susceptible to the growth-suppressive effect of Rac1 inhibition, suggesting that the decrease in "basal" proliferation upon Rac1 inhibition was caused by potentiation of autocrine TGF-beta growth inhibition. CONCLUSIONS: In malignant cells with a functional TGF-beta signalling pathway Rac1 antagonizes the TGF beta1 growth inhibitory response and enhances cell migration by antagonistically regulating Smad2 and Smad3 activation. This study reveals that Rac1 is prooncogenic in that it can alter TGF-beta signalling at the R-Smad level from a tumour-suppressive towards a tumour-promoting outcome. Hence, Rac1 might represent a viable target for therapeutic intervention to inhibit PDAC progression. PMID- 21624124 TI - Parasites in pet reptiles. AB - Exotic reptiles originating from the wild can be carriers of many different pathogens and some of them can infect humans. Reptiles imported into Slovenia from 2000 to 2005, specimens of native species taken from the wild and captive bred species were investigated. A total of 949 reptiles (55 snakes, 331 lizards and 563 turtles), belonging to 68 different species, were examined for the presence of endoparasites and ectoparasites. Twelve different groups (Nematoda (5), Trematoda (1), Acanthocephala (1), Pentastomida (1) and Protozoa (4)) of endoparasites were determined in 26 (47.3%) of 55 examined snakes. In snakes two different species of ectoparasites were also found. Among the tested lizards eighteen different groups (Nematoda (8), Cestoda (1), Trematoda (1), Acanthocephala (1), Pentastomida (1) and Protozoa (6)) of endoparasites in 252 (76.1%) of 331 examined animals were found. One Trombiculid ectoparasite was determined. In 563 of examined turtles eight different groups (Nematoda (4), Cestoda (1), Trematoda (1) and Protozoa (2)) of endoparasites were determined in 498 (88.5%) animals. In examined turtles three different species of ectoparasites were seen. The established prevalence of various parasites in reptiles used as pet animals indicates the need for examination on specific pathogens prior to introduction to owners. PMID- 21624125 TI - Cetrorelix suppresses the preovulatory LH surge and ovulation induced by ovulation-inducing factor (OIF) present in llama seminal plasma. AB - BACKGROUND: The purpose of the study was to determine if the effect of llama OIF on LH secretion is mediated by stimulation of the hypothalamus or pituitary gland. METHODS: Using a 2-by-2 factorial design to examine the effects of OIF vs GnRH with or without a GnRH antagonist, llamas with a growing ovarian follicle greater than or equal to 8 mm were assigned randomly to four groups (n = 7 per group) and a) pre-treated with 1.5 mg of GnRH antagonist (cetrorelix acetate) followed by 1 mg of purified llama OIF, b) pre-treated with 1.5 mg of cetrorelix followed by 50 micrograms of GnRH, c) pre-treated with a placebo (2 ml of saline) followed by 1 mg of purified llama OIF or d) pre-treated with a placebo (2 ml of saline) followed by 50 micrograms of GnRH. Pre-treatment with cetrorelix or saline was given as a single slow intravenous dose 2 hours before intramuscular administration of either GnRH or OIF. Blood samples for LH measurement were taken every 15 minutes from 1.5 hours before to 8 hours after treatment. The ovaries were examined by ultrasonography to detect ovulation and CL formation. Blood samples for progesterone measurement were taken every-other-day from Day 0 (day of treatment) to Day 16. RESULTS: Ovulation rate was not different (P = 0.89) between placebo+GnRH (86%) and placebo+OIF groups (100%); however, no ovulations were detected in llamas pre-treated with cetrorelix. Plasma LH concentrations surged (P < 0.01) after treatment in both placebo+OIF and placebo+GnRH groups, but not in the cetrorelix groups. Maximum plasma LH concentrations and CL diameter profiles did not differ between the placebo-treated groups, but plasma progesterone concentrations were higher (P < 0.05), on days 6, 8 and 12 after treatment, in the OIF- vs GnRH-treated group. CONCLUSION: Cetrorelix (GnRH antagonist) inhibited the preovulatory LH surge induced by OIF in llamas suggesting that LH secretion is modulated by a direct or indirect effect of OIF on GnRH neurons in the hypothalamus. PMID- 21624126 TI - Moving pictures of the human microbiome. AB - BACKGROUND: Understanding the normal temporal variation in the human microbiome is critical to developing treatments for putative microbiome-related afflictions such as obesity, Crohn's disease, inflammatory bowel disease and malnutrition. Sequencing and computational technologies, however, have been a limiting factor in performing dense time series analysis of the human microbiome. Here, we present the largest human microbiota time series analysis to date, covering two individuals at four body sites over 396 timepoints. RESULTS: We find that despite stable differences between body sites and individuals, there is pronounced variability in an individual's microbiota across months, weeks and even days. Additionally, only a small fraction of the total taxa found within a single body site appear to be present across all time points, suggesting that no core temporal microbiome exists at high abundance (although some microbes may be present but drop below the detection threshold). Many more taxa appear to be persistent but non-permanent community members. CONCLUSIONS: DNA sequencing and computational advances described here provide the ability to go beyond infrequent snapshots of our human-associated microbial ecology to high-resolution assessments of temporal variations over protracted periods, within and between body habitats and individuals. This capacity will allow us to define normal variation and pathologic states, and assess responses to therapeutic interventions. PMID- 21624127 TI - Mutation analysis of the WNT4 gene in Han Chinese women with premature ovarian failure. AB - BACKGROUND: The WNT4 gene plays an important role in female sex determination and differentiation. It also contributes to maintaining of the ovaries and the survival of follicles. METHODS: We sequenced the coding region and splice sites of WNT4 in 145 Han Chinese women with premature ovarian failure (POF) and 200 healthy controls. RESULTS: Only one novel variation, in Exon 2 (195C > T), was detected among the women with POF. However, this synonymous variation did not result in a change in amino acid sequence (65 Asp > Asp). No further variants were found in any of the samples. CONCLUSION: Although we cannot provide any evidence that it is a possible disease-causing gene, this study is the first attempt to investigate the possible role of WNT4 in Han Chinese women with POF. PMID- 21624128 TI - Costunolide causes mitotic arrest and enhances radiosensitivity in human hepatocellular carcinoma cells. AB - PURPOSE: This work aimed to investigate the effect of costunolide, a sesquiterpene lactone isolated from Michelia compressa, on cell cycle distribution and radiosensitivity of human hepatocellular carcinoma (HCC) cells. METHODS: The assessment used in this study included: cell viability assay, cell cycle analysis by DNA histogram, expression of phosphorylated histone H3 (Ser 10) by flow cytometer, mitotic index by Liu's stain and morphological observation, mitotic spindle alignment by immunofluorescence of alpha-tubulin, expression of cell cycle-related proteins by Western blotting, and radiation survival by clonogenic assay. RESULTS: Our results show that costunolide reduced the viability of HA22T/VGH cells. It caused a rapid G2/M arrest at 4 hours shown by DNA histogram. The increase in phosphorylated histone H3 (Ser 10)-positive cells and mitotic index indicates costunolide-treated cells are arrested at mitosis, not G2, phase. Immunofluorescence of alpha-tubulin for spindle formation further demonstrated these cells are halted at metaphase. Costunolide up-regulated the expression of phosphorylated Chk2 (Thr 68), phosphorylated Cdc25c (Ser 216), phosphorylated Cdk1 (Tyr 15) and cyclin B1 in HA22T/VGH cells. At optimal condition causing mitotic arrest, costunolide sensitized HA22T/VGH HCC cells to ionizing radiation with sensitizer enhancement ratio up to 1.9. CONCLUSIONS: Costunolide could reduce the viability and arrest cell cycling at mitosis in hepatoma cells. Logical exploration of this mitosis-arresting activity for cancer therapeutics shows costunolide enhanced the killing effect of radiotherapy against human HCC cells. PMID- 21624129 TI - Dual function of polycomb group proteins in differentiated murine T helper (CD4+) cells. AB - BACKGROUND: Following antigen recognition, naive T helper (Th; CD4+) cells can differentiate toward one of several effector lineages such as Th1 and Th2; each expressing distinctive transcriptional profiles of cytokine genes. These cytokines eventually instruct the strategy of the immune response. In our search for factors that propagate the transcriptional programs of differentiated Th cells, we previously found that Polycomb group (PcG) proteins, which are known as epigenetic regulators that maintain repressive chromatin states, bind differentially the signature cytokine genes. Unexpectedly, their binding to the Ifng (Interferon-g) in Th1 cells and Il4 (Interleukin-4) in Th2 cells, was correlated with transcriptional activation. Therefore, in this study we aimed to determine the functional role of PcG proteins in the regulation of the expression of the signature cytokine genes. METHODS: PcG proteins were knocked down in primary and established murine Th cells using transduction of lentiviruses encoding short hairpin RNAs (shRNAs) directed to Mel-18, Ezh2, Eed and Ring1A, representative of two different PcG complexes. The chromatin structure and the binding activity of PcG proteins and transcription factors at the Ifng promoter were assessed by chromatin immunoprecipitation (ChIP) assays. RESULTS: Downregulation of PcG proteins was consistent with their function as positive regulators of the signature cytokine genes in primary and established Th1 and Th2 cells. Moreover, the PcG protein Mel-18 was necessary to recruit the Th1-lineage specifying transcription factor T-bet, and the T cell receptor (TCR)-inducible transcription factor NFAT1 to the Ifng promoter in Th1 cells. Nevertheless, our results suggest that PcG proteins can function also as conventional transcriptional repressors in Th cells of their known target the Hoxa7 gene. CONCLUSIONS: Our data support a model whereby the non-differentially expressed PcG proteins are recruited in a Th-lineage specific manner to their target genes to enforce the maintenance of specific transcriptional programs as transcriptional repressors or activators. Although our results suggest a direct effect of PcG proteins in the regulation of cytokine gene expression, indirect functions cannot be excluded. PMID- 21624130 TI - A novel candidate HIV vaccine vector based on the replication deficient Capripoxvirus, Lumpy skin disease virus (LSDV). AB - BACKGROUND: The Capripoxvirus, Lumpy skin disease virus (LSDV) has a restricted host-range and is being investigated as a novel HIV-1 vaccine vector. LSDV does not complete its replication cycle in non-ruminant hosts. METHODS: The safety of LSDV was tested at doses of 104 and 106 plaque forming units in two strains of immunocompromised mice, namely RAG mice and CD4 T cell knockout mice. LSDV expressing HIV-1 subtype C Gag, reverse transcriptase (RT), Tat and Nef as a polyprotein (Grttn), (rLSDV-grttn), was constructed. The immunogenicity of rLSDV grttn was tested in homologous prime-boost regimens as well as heterologous prime boost regimes in combination with a DNA vaccine (pVRC-grttn) or modified vaccinia Ankara vaccine (rMVA-grttn) both expressing Grttn. RESULTS: Safety was demonstrated in two strains of immunocompromised mice.In the immunogenicity experiments mice developed high magnitudes of HIV-specific cells producing IFN gamma and IL-2. A comparison of rLSDV-grttn and rMVA-grttn to boost a DNA vaccine (pVRC-grttn) indicated a DNA prime and rLSDV-grttn boost induced a 2 fold (p < 0.01) lower cumulative frequency of Gag- and RT-specific IFN-gamma CD8 and CD4 cells than a boost with rMVA-grttn. However, the HIV-specific cells induced by the DNA vaccine prime rLSDV-grttn boost produced greater than 3 fold (p < 0.01) more IFN- gamma than the HIV-specific cells induced by the DNA vaccine prime rMVA grttn boost. A boost of HIV-specific CD4 cells producing IL-2 was only achieved with the DNA vaccine prime and rLSDV-grttn boost. Heterologous prime-boost combinations of rLSDV-grttn and rMVA-grttn induced similar cumulative frequencies of IFN- gamma producing Gag- and RT-specific CD8 and CD4 cells. A significant difference (p < 0.01) between the regimens was the higher capacity (2.1 fold) of Gag-and RT-specific CD4 cells to produce IFN-gamma with a rMVA-grttn prime - rLSDV-grttn boost. This regimen also induced a 1.5 fold higher (p < 0.05) frequency of Gag- and RT-specific CD4 cells producing IL-2. CONCLUSIONS: LSDV was demonstrated to be non-pathogenic in immunocompromised mice. The rLSDV-grttn vaccine was immunogenic in mice particularly in prime-boost regimens. The data suggests that this novel vaccine may be useful for enhancing, in particular, HIV specific CD4 IFN- gamma and IL-2 responses induced by a priming vaccine. PMID- 21624131 TI - Analysis of timeliness of infectious disease reporting in the Netherlands. AB - BACKGROUND: Timely reporting of infectious disease cases to public health authorities is essential to effective public health response. To evaluate the timeliness of reporting to the Dutch Municipal Health Services (MHS), we used as quantitative measures the intervals between onset of symptoms and MHS notification, and between laboratory diagnosis and notification with regard to six notifiable diseases. METHODS: We retrieved reporting data from June 2003 to December 2008 from the Dutch national notification system for shigellosis, EHEC/STEC infection, typhoid fever, measles, meningococcal disease, and hepatitis A virus (HAV) infection. For each disease, median intervals between date of onset and MHS notification were calculated and compared with the median incubation period. The median interval between date of laboratory diagnosis and MHS notification was similarly analysed. For the year 2008, we also investigated whether timeliness is improved by MHS agreements with physicians and laboratories that allow direct laboratory reporting. Finally, we investigated whether reports made by post, fax, or e-mail were more timely. RESULTS: The percentage of infectious diseases reported within one incubation period varied widely, between 0.4% for shigellosis and 90.3% for HAV infection. Not reported within two incubation periods were 97.1% of shigellosis cases, 76.2% of cases of EHEC/STEC infection, 13.3% of meningococcosis cases, 15.7% of measles cases, and 29.7% of typhoid fever cases. A substantial percentage of infectious disease cases was reported more than three days after laboratory diagnosis, varying between 12% for meningococcosis and 42% for shigellosis. MHS which had agreements with physicians and laboratories showed a significantly shorter notification time compared to MHS without such agreements. CONCLUSIONS: Over the study period, many cases of the six notifiable diseases were not reported within two incubation periods, and many were reported more than three days after laboratory diagnosis. An increase in direct laboratory reporting of diagnoses to MHS would improve timeliness, as would the use of fax rather than post or e-mail. Automated reporting systems have to be explored in the Netherlands. Development of standardised and improved measures for timeliness is needed. PMID- 21624132 TI - The role of traditional healers in tooth extractions in Lekie Division, Cameroon. AB - BACKGROUND: The extraction of the teeth by traditional healers in Cameroon is an established cultural practice in the central region of the Cameroon. Traditional healers (TH) use herbs and crude un-sterilized instruments and tools for the tooth extraction procedure. The present study investigates the knowledge and practices of traditional healers regarding tooth extraction and the management of its complications. METHODS: A cross sectional design utilizing semi-structured questionnaires was used to collect the data from traditional healers and their patients. RESULTS: Sixteen traditional healers (TH) were interviewed. All were male and the majority were between 25-35 years old. The most important reason given for the removal of a tooth was "if it has a hole". All reported using herbs to control bleeding and pain after extractions. Only 20% used gloves between patients when extracting a tooth and just over a third (31.3%) gave post operative instructions. Eighty seven percent managed complications with herbs and 62.5% reported that they would refer their patients to a dentist whenever there are complications. Only a third (31.3%) was familiar with the basic anatomy of a tooth and more than half (56.3%) reported that tooth extractions are the only treatment for dental problems.One hundred and fifty patients were interviewed with a mean age of 29 years. More than two thirds were in the 21-30 year age group and just over half were male. Sixty six percent reported that they visited the TH because it is cheap, 93.3% were satisfied with the treatment they received while 95.3% reported said they never had a problem after an extraction. CONCLUSIONS: Tooth extractions using medicinal plants is well established in Lekie division, Cameroon. Infection control during extraction is not the norm. Traditional healers are willing to co-operate with oral health workers in improving the oral health of their patients. Mutual cooperation, collaboration and integrating TH into primary oral health care services need to be increased. PMID- 21624133 TI - Alteration of T cell cytokine production in PLPp-139-151-induced EAE in SJL mice by an immunostimulatory CpG oligonucleotide. AB - Experimental autoimmune encephalomyelitis (EAE) is--in certain aspects--regarded as an animal model of the human CNS autoimmune disease multiple sclerosis (MS). While in EAE CNS-autoantigen-specific immunity is induced in a defined way, the initial processes leading to CNS autoimmunity in humans are so far unknown. Despite essential restrictions, which exist regarding the interpretation of EAE data towards MS, EAE might be a useful model to study certain basic aspects of CNS autoimmunity. Studies in MS have demonstrated that established autoimmune pathology can be critically influenced by environmental factors, in particular viral and bacterial infections. To investigate this interaction, EAE as an instrument to study CNS autoimmunity under defined conditions appears to be a suitable experimental tool. For this reason, we here investigated the influence of the Toll-like-receptor (TLR) ligand CpG oligonucleotide (CpG) on already established CNS autoimmunity in murine proteolipid protein (PLP)-induced EAE in SJL mice. CpG were found to co-stimulate PLPp-specific IFN-gamma production in the peripheral immune system and in the CNS. However, CpG induced Interleukin (IL)-17 production in the inflamed CNS both alone and in combination with additional PLPp stimulation. These findings might indicate a mechanism by which systemic infections and the microbial stimuli associated with them may influence already existing CNS autoimmune pathology. PMID- 21624134 TI - Meta-analysis of microarray data using a pathway-based approach identifies a 37 gene expression signature for systemic lupus erythematosus in human peripheral blood mononuclear cells. AB - BACKGROUND: A number of publications have reported the use of microarray technology to identify gene expression signatures to infer mechanisms and pathways associated with systemic lupus erythematosus (SLE) in human peripheral blood mononuclear cells. However, meta-analysis approaches with microarray data have not been well-explored in SLE. METHODS: In this study, a pathway-based meta analysis was applied to four independent gene expression oligonucleotide microarray data sets to identify gene expression signatures for SLE, and these data sets were confirmed by a fifth independent data set. RESULTS: Differentially expressed genes (DEGs) were identified in each data set by comparing expression microarray data from control samples and SLE samples. Using Ingenuity Pathway Analysis software, pathways associated with the DEGs were identified in each of the four data sets. Using the leave one data set out pathway-based meta-analysis approach, a 37-gene metasignature was identified. This SLE metasignature clearly distinguished SLE patients from controls as observed by unsupervised learning methods. The final confirmation of the metasignature was achieved by applying the metasignature to a fifth independent data set. CONCLUSIONS: The novel pathway based meta-analysis approach proved to be a useful technique for grouping disparate microarray data sets. This technique allowed for validated conclusions to be drawn across four different data sets and confirmed by an independent fifth data set. The metasignature and pathways identified by using this approach may serve as a source for identifying therapeutic targets for SLE and may possibly be used for diagnostic and monitoring purposes. Moreover, the meta-analysis approach provides a simple, intuitive solution for combining disparate microarray data sets to identify a strong metasignature. PMID- 21624135 TI - Histone deacetylase turnover and recovery in sulforaphane-treated colon cancer cells: competing actions of 14-3-3 and Pin1 in HDAC3/SMRT corepressor complex dissociation/reassembly. AB - BACKGROUND: Histone deacetylase (HDAC) inhibitors are currently undergoing clinical evaluation as anti-cancer agents. Dietary constituents share certain properties of HDAC inhibitor drugs, including the ability to induce global histone acetylation, turn-on epigenetically-silenced genes, and trigger cell cycle arrest, apoptosis, or differentiation in cancer cells. One such example is sulforaphane (SFN), an isothiocyanate derived from the glucosinolate precursor glucoraphanin, which is abundant in broccoli. Here, we examined the time-course and reversibility of SFN-induced HDAC changes in human colon cancer cells. RESULTS: Cells underwent progressive G2/M arrest over the period 6-72 h after SFN treatment, during which time HDAC activity increased in the vehicle-treated controls but not in SFN-treated cells. There was a time-dependent loss of class I and selected class II HDAC proteins, with HDAC3 depletion detected ahead of other HDACs. Mechanism studies revealed no apparent effect of calpain, proteasome, protease or caspase inhibitors, but HDAC3 was rescued by cycloheximide or actinomycin D treatment. Among the protein partners implicated in the HDAC3 turnover mechanism, silencing mediator for retinoid and thyroid hormone receptors (SMRT) was phosphorylated in the nucleus within 6 h of SFN treatment, as was HDAC3 itself. Co-immunoprecipitation assays revealed SFN-induced dissociation of HDAC3/SMRT complexes coinciding with increased binding of HDAC3 to 14-3-3 and peptidyl-prolyl cis/trans isomerase 1 (Pin1). Pin1 knockdown blocked the SFN induced loss of HDAC3. Finally, SFN treatment for 6 or 24 h followed by SFN removal from the culture media led to complete recovery of HDAC activity and HDAC protein expression, during which time cells were released from G2/M arrest. CONCLUSION: The current investigation supports a model in which protein kinase CK2 phosphorylates SMRT and HDAC3 in the nucleus, resulting in dissociation of the corepressor complex and enhanced binding of HDAC3 to 14-3-3 or Pin1. In the cytoplasm, release of HDAC3 from 14-3-3 followed by nuclear import is postulated to compete with a Pin1 pathway that directs HDAC3 for degradation. The latter pathway predominates in colon cancer cells exposed continuously to SFN, whereas the former pathway is likely to be favored when SFN has been removed within 24 h, allowing recovery from cell cycle arrest. PMID- 21624136 TI - Donnan effect on chloride ion distribution as a determinant of body fluid composition that allows action potentials to spread via fast sodium channels. AB - Proteins in any solution with a pH value that differs from their isoelectric point exert both an electric Donnan effect (DE) and colloid osmotic pressure. While the former alters the distribution of ions, the latter forces water diffusion. In cells with highly Cl--permeable membranes, the resting potential is more dependent on the cytoplasmic pH value, which alters the Donnan effect of cell proteins, than on the current action of Na/K pumps. Any weak (positive or negative) electric disturbances of their resting potential are quickly corrected by chloride shifts.In many excitable cells, the spreading of action potentials is mediated through fast, voltage-gated sodium channels. Tissue cells share similar concentrations of cytoplasmic proteins and almost the same exposure to the interstitial fluid (IF) chloride concentration. The consequence is that similar intra- and extra-cellular chloride concentrations make these cells share the same Nernst value for Cl-.Further extrapolation indicates that cells with the same chloride Nernst value and high chloride permeability should have similar resting membrane potentials, more negative than -80 mV. Fast sodium channels require potassium levels >20 times higher inside the cell than around it, while the concentration of Cl- ions needs to be >20 times higher outside the cell.When osmotic forces, electroneutrality and other ions are all taken into account, the overall osmolarity needs to be near 280 to 300 mosm/L to reach the required resting potential in excitable cells. High plasma protein concentrations keep the IF chloride concentration stable, which is important in keeping the resting membrane potential similar in all chloride-permeable cells. Probable consequences of this concept for neuron excitability, erythrocyte membrane permeability and several features of circulation design are briefly discussed. PMID- 21624137 TI - Formant analysis in dysphonic patients and automatic Arabic digit speech recognition. AB - BACKGROUND AND OBJECTIVE: There has been a growing interest in objective assessment of speech in dysphonic patients for the classification of the type and severity of voice pathologies using automatic speech recognition (ASR). The aim of this work was to study the accuracy of the conventional ASR system (with Mel frequency cepstral coefficients (MFCCs) based front end and hidden Markov model (HMM) based back end) in recognizing the speech characteristics of people with pathological voice. MATERIALS AND METHODS: The speech samples of 62 dysphonic patients with six different types of voice disorders and 50 normal subjects were analyzed. The Arabic spoken digits were taken as an input. The distribution of the first four formants of the vowel /a/ was extracted to examine deviation of the formants from normal. RESULTS: There was 100% recognition accuracy obtained for Arabic digits spoken by normal speakers. However, there was a significant loss of accuracy in the classifications while spoken by voice disordered subjects. Moreover, no significant improvement in ASR performance was achieved after assessing a subset of the individuals with disordered voices who underwent treatment. CONCLUSION: The results of this study revealed that the current ASR technique is not a reliable tool in recognizing the speech of dysphonic patients. PMID- 21624138 TI - Goal-directed haemodynamic therapy during elective total hip arthroplasty under regional anaesthesia. AB - INTRODUCTION: Total hip replacement is one of the most commonly performed major orthopaedic operations. Goal-directed therapy (GDT) using haemodynamic monitoring has previously demonstrated outcome benefits in high-risk surgical patients under general anaesthesia. GDT has never been formally assessed during regional anaesthesia. METHODS: Patients undergoing total hip replacement while under regional anaesthesia were randomised to either the control group (CTRL) or the protocol group (GDT). Patients in the GDT group, in addition to standard monitoring, were connected to the FloTrac sensor/Vigileo monitor haemodynamic monitoring system, and a GDT protocol was used to maximise the stroke volume and target the oxygen delivery index to > 600 mL/minute/m2. RESULTS: Patients randomised to the GDT group were given a greater volume of intravenous fluids during the intraoperative period (means +/- standard deviation (SD): 6,032 +/- 1,388 mL vs. 2,635 +/- 346 mL; P < 0.0001), and more of the GDT patients received dobutamine (0 of 20 CTRL patients vs. 11 of 20 GDT patients; P < 0.0003). The GDT patients also received more blood transfused during the intraoperative period (means +/- SD: 595 +/- 316 mL vs. 0 +/- 0 mL; P < 0.0001), although the CTRL group received greater volumes of blood replacement postoperatively (CTRL patients 658 +/- 68 mL vs. GDT patients 198 +/- 292 mL; P < 0.001). Overall blood consumption (intraoperatively and postoperatively) was not different between the two groups. There were an increased number of complications in the CTRL group (20 of 20 CTRL patients (100%) vs. 16 of 20 GDT patients (80%); P = 0.05). These outcomes were predominantly due to a difference in minor complications (20 of 20 CTRL patients (100%) vs. 15 of 20 GDT patients (75%); P = 0.047). CONCLUSIONS: GDT applied during regional anaesthesia in patients undergoing elective total hip replacement changes intraoperative fluid management and may improve patient outcomes by decreasing postoperative complications. Larger trials are required to confirm our findings. PMID- 21624139 TI - Temperature and time stability of whole blood lactate: implications for feasibility of pre-hospital measurement. AB - BACKGROUND: To determine the time and temperature stability of whole blood lactate using experimental conditions applicable to the out-of-hospital environment. FINDINGS: We performed a prospective, clinical laboratory-based study at an academic hospital. Whole blood lactate was obtained by venipuncture from five post-prandial, resting subjects. Blood was stored in lithium heparinized vacutainers in three temperature conditions: 1) room temperature (20 degrees C), 2) wrapped in a portable, instant ice pack (0 degrees C), or 3) wet ice (0 degrees C). Lactate concentrations (mmol/L) were measured at 0, 5, 10, 20, and 30 minutes after sampling, and compared using repeated measures analysis of variance.Mean baseline lactate among resting subjects (N = 5) was 1.24 mmol/L (95%CI: 0.49,1.98 mmol/L). After 30 minutes, lactate concentration increased, on average, by 0.08 mmol/L (95%CI: 0.02,0.13 mmol/L), 0.18 mmol/L (95%CI: 0.07,0.28 mmol/L), and 0.36 mmol/L (95%CI: 0.24,0.47 mmol/L) when stored in wet ice, ice pack, and room temperature, respectively. The increase in lactate was similar in samples wrapped in portable ice pack or stored in wet ice at all time points (p > 0.05), and met criteria for equivalence at 30 minutes. However, lactate measurements from whole blood stored at room temperature were significantly greater, on average, than wet ice or portable ice pack within five and ten minutes, respectively (p < 0.05). CONCLUSIONS: Whole blood lactate measurements using samples stored in a portable ice pack are similar to wet ice for up to 30 minutes. These conditions are applicable to the out-of-hospital environment, and should inform future studies of pre-hospital measurement of lactate. PMID- 21624141 TI - A robust approach based on Weibull distribution for clustering gene expression data. AB - BACKGROUND: Clustering is a widely used technique for analysis of gene expression data. Most clustering methods group genes based on the distances, while few methods group genes according to the similarities of the distributions of the gene expression levels. Furthermore, as the biological annotation resources accumulated, an increasing number of genes have been annotated into functional categories. As a result, evaluating the performance of clustering methods in terms of the functional consistency of the resulting clusters is of great interest. RESULTS: In this paper, we proposed the WDCM (Weibull Distribution based Clustering Method), a robust approach for clustering gene expression data, in which the gene expressions of individual genes are considered as the random variables following unique Weibull distributions. Our WDCM is based on the concept that the genes with similar expression profiles have similar distribution parameters, and thus the genes are clustered via the Weibull distribution parameters. We used the WDCM to cluster three cancer gene expression data sets from the lung cancer, B-cell follicular lymphoma and bladder carcinoma and obtained well-clustered results. We compared the performance of WDCM with k-means and Self Organizing Map (SOM) using functional annotation information given by the Gene Ontology (GO). The results showed that the functional annotation ratios of WDCM are higher than those of the other methods. We also utilized the external measure Adjusted Rand Index to validate the performance of the WDCM. The comparative results demonstrate that the WDCM provides the better clustering performance compared to k-means and SOM algorithms. The merit of the proposed WDCM is that it can be applied to cluster incomplete gene expression data without imputing the missing values. Moreover, the robustness of WDCM is also evaluated on the incomplete data sets. CONCLUSIONS: The results demonstrate that our WDCM produces clusters with more consistent functional annotations than the other methods. The WDCM is also verified to be robust and is capable of clustering gene expression data containing a small quantity of missing values. PMID- 21624140 TI - Chronic granulomatous disease: a review of the infectious and inflammatory complications. AB - Chronic Granulomatous Disease is the most commonly encountered immunodeficiency involving the phagocyte, and is characterized by repeated infections with bacterial and fungal pathogens, as well as the formation of granulomas in tissue. The disease is the result of a disorder of the NADPH oxidase system, culminating in an inability of the phagocyte to generate superoxide, leading to the defective killing of pathogenic organisms. This can lead to infections with Staphylococcus aureus, Psedomonas species, Nocardia species, and fungi (such as Aspergillus species and Candida albicans). Involvement of vital or large organs can contribute to morbidity and/or mortality in the affected patients. Major advances have occurred in the diagnosis and treatment of this disease, with the potential for gene therapy or stem cell transplantation looming on the horizon. PMID- 21624142 TI - HEART: heart exercise and remote technologies: a randomized controlled trial study protocol. AB - BACKGROUND: Cardiovascular disease (CVD) is the leading cause of death worldwide. Cardiac rehabilitation (CR) is aimed at improving health behaviors to slow or reverse the progression of CVD disease. Exercise is a central element of CR. Technologies such as mobile phones and the Internet (mHealth) offer potential to overcome many of the psychological, physical, and geographical barriers that have been associated with lack of participation in exercise-based CR. We aim to trial the effectiveness of a mobile phone delivered exercise-based CR program to increase exercise capacity and functional outcomes compared with usual CR care in adults with CVD. This paper outlines the rationale and methods of the trial. METHODS: A single-blinded parallel two-arm randomized controlled trial is being conducted. A total of 170 people will be randomized at 1:1 ratio either to receive a mHealth CR program or usual care. Participants are identified by CR nurses from two metropolitan hospitals in Auckland, New Zealand through outpatient clinics and existing databases. Consenting participants are contacted to attend a baseline assessment. The intervention consists of a theory-based, personalized, automated package of text and video message components via participants' mobile phones and the Internet to increase exercise behavior, delivered over six months. The control group will continue with usual CR. Data collection occurs at baseline and 24 weeks (post-intervention). The primary outcome is change in maximal oxygen uptake from baseline to 24 weeks. Secondary outcomes include post-intervention measures on self-reported physical activity (IPAQ), cardiovascular risk factors (systolic blood pressure, weight, and waist to hip ratio), health related quality of life (SF-36), and cost-effectiveness. DISCUSSION: This manuscript presents the protocol for a randomized controlled trial of a mHealth exercise-based CR program. Results of this trial will provide much needed information about physical and psychological well-being, and cost effectiveness of an automated telecommunication intervention. If effective, this intervention has enormous potential to improve the delivery of CR and could easily be scaled up to be delivered nationally (and internationally) in a very short time, enhancing the translational aspect of this research. It also has potential to extend to comprehensive CR (nutrition advice, smoking cessation, medication adherence). TRIAL REGISTRATION: ACTRN12611000117910. PMID- 21624143 TI - SO2426 is a positive regulator of siderophore expression in Shewanella oneidensis MR-1. AB - BACKGROUND: The Shewanella oneidensis MR-1 genome encodes a predicted orphan DNA binding response regulator, SO2426. Previous studies with a SO2426-deficient MR-1 strain suggested a putative functional role for SO2426 in the regulation of iron acquisition genes, in particular, the siderophore (hydroxamate) biosynthesis operon so3030-3031-3032. To further investigate the functional role of SO2426 in iron homeostasis, we employed computational strategies to identify putative gene targets of SO2426 regulation and biochemical approaches to validate the participation of SO2426 in the control of siderophore biosynthesis in S. oneidensis MR-1. RESULTS: In silico prediction analyses revealed a single 14-bp consensus motif consisting of two tandem conserved pentamers (5'-CAAAA-3') in the upstream regulatory regions of 46 genes, which were shown previously to be significantly down-regulated in a so2426 deletion mutant. These genes included so3030 and so3032, members of an annotated siderophore biosynthetic operon in MR 1. Electrophoretic mobility shift assays demonstrated that the SO2426 protein binds to its motif in the operator region of so3030. A "short" form of SO2426, beginning with a methionine at position 11 (M11) of the originally annotated coding sequence for SO2426, was also functional in binding to its consensus motif, confirming previous 5' RACE results that suggested that amino acid M11 is the actual translation start codon for SO2426. Alignment of SO2426 orthologs from all sequenced Shewanella spp. showed a high degree of sequence conservation beginning at M11, in addition to conservation of a putative aspartyl phosphorylation residue and the helix-turn-helix (HTH) DNA-binding domain. Finally, the so2426 deletion mutant was unable to synthesize siderophores at wild type rates upon exposure to the iron chelator 2,2'-dipyridyl. CONCLUSIONS: Collectively, these data support the functional characterization of SO2426 as a positive regulator of siderophore-mediated iron acquisition and provide the first insight into a coordinate program of multiple regulatory schemes controlling iron homeostasis in S. oneidensis MR-1. PMID- 21624144 TI - Engineering Klebsiella sp. 601 multicopper oxidase enhances the catalytic efficiency towards phenolic substrates. AB - BACKGROUND: Structural comparison between bacterial CueO and fungal laccases has suggested that a charged residue Glu (E106) in CueO replaces the corresponding residue Phe in fungal laccases at the gate of the tunnel connecting type II copper to the protein surface and an extra alpha-helix (L351-G378) near the type I copper site covers the substrate binding pocket and might compromise the electron transfer from substrate to type I copper. To test this hypothesis, several mutants were made in Klebsiella sp. 601 multicopper oxidase, which is highly homologous to E. coli CueO with a similarity of 90% and an identity of 78%. RESULTS: The E106F mutant gave smaller K(m) (2.4-7 fold) and k(cat) (1-4.4 fold) values for all three substrates DMP, ABTS and SGZ as compared with those for the wild-type enzyme. Its slightly larger k(cat)/K(m) values for three substrates mainly come from the decreased K(m). Deleting alpha-helix (L351-G378) resulted in the formation of inactive inclusion body when the mutant (Delta)alpha351-378 was expressed in E. coli. Another mutant alpha351-380M was then made via substitution of seven amino acid residues in the alpha-helix (L351 G378) region. The alpha351-380M mutant was active, and displayed a far-UV CD spectrum markedly different from that for wild-type enzyme. Kinetic studies showed the alpha351-380M mutant gave very low K(m) values for DMP, ABTS and SGZ, 4.5-, 1.9- and 7-fold less than those for the wild type. In addition, k(cat)/K(m) values were increased, 9.4-fold for DMP, similar for ABTS and 3-fold for SGZ. CONCLUSION: The Glu residue at position 106 appears not to be the only factor affecting the copper binding, and it may also play a role in maintaining enzyme conformation. The alpha-helix (L351-G378) may not only block access to the type I copper site but also play a role in substrate specificities of bacterial MCOs. The alpha351-380M mutant catalyzing oxidation of the phenolic substrate DMP effectively would be very useful in green chemistry. PMID- 21624145 TI - Chemical modification of L-glutamine to alpha-amino glutarimide on autoclaving facilitates Agrobacterium infection of host and non-host plants: A new use of a known compound. AB - BACKGROUND: Accidental autoclaving of L-glutamine was found to facilitate the Agrobacterium infection of a non host plant like tea in an earlier study. In the present communication, we elucidate the structural changes in L-glutamine due to autoclaving and also confirm the role of heat transformed L-glutamine in Agrobacterium mediated genetic transformation of host/non host plants. RESULTS: When autoclaved at 121 degrees C and 15 psi for 20 or 40 min, L-glutamine was structurally modified into 5-oxo proline and 3-amino glutarimide (alpha-amino glutarimide), respectively. Of the two autoclaved products, only alpha-amino glutarimide facilitated Agrobacterium infection of a number of resistant to susceptible plants. However, the compound did not have any vir gene inducing property. CONCLUSIONS: We report a one pot autoclave process for the synthesis of 5-oxo proline and alpha-amino glutarimide from L-glutamine. Xenobiotic detoxifying property of alpha-amino glutarimide is also proposed. PMID- 21624146 TI - Introduction: Mentholated cigarettes and public health. PMID- 21624147 TI - Epidemiology of menthol cigarette use in the United States. AB - BACKGROUND: Approximately one-fourth of all cigarettes sold in the United States have the descriptor "menthol" on the cigarette pack. It is important to determine what socio-demographic factors are associated with smoking menthol cigarettes if indeed these types of cigarettes are related to smoking initiation, higher exposure to smoke constituents, nicotine dependence, or reduced smoking cessation. METHODS: The National Cancer Institute (NCI) conducted a review of the scientific literature on this topic which we completed by adding more recently published articles via PubMed. We also conducted further data analyses using the National Survey on Drug Use and Health, the National Youth Tobacco Survey, the Monitoring the Future Survey, and the National Health and Nutrition Examination Survey to provide up-to-date information on this topic. RESULTS: Menthol cigarettes are disproportionately smoked by adolescents, blacks/African Americans, adult females, those living in the Northeast of the United States and those with family incomes lower than $50,000. Based on self-reports of menthol cigarette use, menthol cigarette use among smokers have increased from 2004 to 2008. However, no increase was observed during these years for predominantly menthol brands like NewportTM, Kool,TM and SalemTM, however, this lack of significant trend may be due, at least in part, due to smaller numbers of smokers of specific brands or sub-brands, which provide estimates which are less precise. CONCLUSION: Menthol cigarettes are disproportionately smoked by groups of U.S. cigarette smokers. It is likely that other disparities in menthol cigarette use exist that we have not covered or have not been studied yet. PMID- 21624148 TI - Marketing of menthol cigarettes and consumer perceptions. AB - In order to more fully understand why individuals smoke menthol cigarettes, it is important to understand the perceptions held by youth and adults regarding menthol cigarettes. Perceptions are driven by many factors, and one factor that can be important is marketing. This review seeks to examine what role, if any, the marketing of menthol cigarettes plays in the formation of consumer perceptions of menthol cigarettes. The available literature suggests that menthol cigarettes may be perceived as safer choices than non-menthol cigarettes. Furthermore, there is significant overlap between menthol cigarette advertising campaigns and the perceptions of these products held by consumers. The marketing of menthol cigarettes has been higher in publications and venues whose target audiences are Blacks/African Americans. Finally, there appears to have been changes in cigarette menthol content over the past decade, which has been viewed by some researchers as an effort to attract different types of smokers. PMID- 21624149 TI - Sensory properties of menthol and smoking topography. AB - Although there is a great deal known about menthol as a flavoring agent in foods and confections, less is known about the particular sensory properties of menthol cigarette smoke. Similarly, although smoking topography (the unique way an individual smokes a cigarette) has been well studied using non-menthol cigarettes, there is relatively less known about how menthol affects smoking behavior. The objective of this review is to assess the sensory properties of menthol tobacco smoke, and smoking topography associated with menthol cigarettes. The cooling, analgesic, taste, and respiratory effects of menthol are well established, and studies have indicated that menthol's sensory attributes can have an influence on the positive, or rewarding, properties associated smoking, including ratings of satisfaction, taste, perceived smoothness, and perceived irritation. Despite these sensory properties, the data regarding menthol's effect on smoking topography are inconsistent. Many of the topography studies have limitations due to various methodological issues. PMID- 21624150 TI - Menthol and initiation of cigarette smoking. AB - The use of tobacco products would not continue without the initiation of their use by youth and adults. Since the vast majority of cigarette smokers begin smoking by age 25, understanding the role of menthol cigarettes in the initiation of smoking in youth (under the age of 18) and young adults (aged 18-25) is especially relevant. Data demonstrate that menthol cigarettes are disproportionately used by youth and young adults. This review seeks to examine what role, if any, menthol plays in the initiation of cigarette smoking. Overall, there is a paucity of data on this topic. The data that do exist suggests that youth who have smoked for less than 1 year are more likely to smoke menthol cigarettes than youth who have smoked for more than 1 year. A lack of data prevents further conclusions on the role of menthol cigarettes in the initiation of smoking. PMID- 21624151 TI - Menthol cigarette smoking and nicotine dependence. AB - Since tobacco use is driven by dependence on nicotine, the primary addictive substance in tobacco, much research has focused on nicotine dependence. Less well understood, however, is the role that menthol plays in nicotine dependence. This review seeks to examine what role, if any, menthol plays in nicotine addiction in adults and youth. Based on research examining several indicators of heaviness of nicotine addiction, including time to first cigarette upon waking, night waking to smoke, as well as some other indications of dependence, it is suggested that menthol cigarette smokers are more heavily dependent on nicotine. Although other indicators of nicotine dependence, including number of cigarettes per day and the Fagerstrom Test of Nicotine Dependence, failed to consistently differentiate menthol and non-menthol smokers, these indicators are thought to be less robust than time to first cigarette. Therefore, though limited, the existing literature suggests that menthol smokers may be more dependence on nicotine. PMID- 21624152 TI - Menthol cigarettes and smoking cessation behavior. AB - Although much is known about smoking cessation behavior, the vast majority of research has not assessed menthol as an independent factor. The objective of this review is to assess the effects, if any, that use of menthol cigarettes has on smoking cessation success in adults and youth. A total of 20 articles are included in this review. Although some studies have found that menthol smokers have less success in quitting smoking, others fail to find significant differences between menthol and non-menthol smokers. Some clinical trials evaluating the efficacy of various cessation treatments have suggested that menthol smokers have poorer outcomes, however two secondary data analysis studies (which used the same original dataset) failed to find any difference in success rate associated with particular treatments. Although there is some suggestion that smoking menthol cigarettes is associated with worse cessation outcomes, differences are not always found. However, if there was a difference, it was always in the direction of worse outcomes for menthol smokers. Given that Black/African American smokers prefer menthol cigarettes more than White smokers, possible interactions with race/ethnicity are discussed. PMID- 21624153 TI - The health effects of menthol cigarettes as compared to non-menthol cigarettes. AB - Since the 1920s, menthol has been added to cigarettes and used as a characterizing flavor. The health effects of cigarette smoking are well documented, however the health effects of menthol cigarettes as compared to non menthol cigarettes is less well studied. This review discusses menthol's effects on 1) biomarkers of tobacco smoke exposure, 2) toxicity and cellular effects, 3) lung function and respiration, 4) pulmonary and/or vascular function, 5) allergic reactions and inflammation, and 6) tobacco-related diseases. It is concluded that menthol is a biologically active compound that has effects by itself and in conjunction with nicotine, however much of the data on the other areas of interest are inconclusive and firm conclusions cannot be drawn. PMID- 21624154 TI - Selected papers from the 13th Annual Bio-Ontologies Special Interest Group Meeting. AB - Over the years, the Bio-Ontologies SIG at ISMB has provided a forum for discussion of the latest and most innovative research in the application of ontologies and more generally the organisation, presentation and dissemination of knowledge in biomedicine and the life sciences. The ten papers selected for this supplement are extended versions of the original papers presented at the 2010 SIG. The papers span a wide range of topics including practical solutions for data and knowledge integration for translational medicine, hypothesis based querying , understanding kidney and urinary pathways, mining the pharmacogenomics literature; theoretical research into the orthogonality of biomedical ontologies, the representation of diseases, the representation of research hypotheses, the combination of ontologies and natural language processing for an annotation framework, the generation of textual definitions, and the discovery of gene interaction networks. PMID- 21624155 TI - The Translational Medicine Ontology and Knowledge Base: driving personalized medicine by bridging the gap between bench and bedside. AB - BACKGROUND: Translational medicine requires the integration of knowledge using heterogeneous data from health care to the life sciences. Here, we describe a collaborative effort to produce a prototype Translational Medicine Knowledge Base (TMKB) capable of answering questions relating to clinical practice and pharmaceutical drug discovery. RESULTS: We developed the Translational Medicine Ontology (TMO) as a unifying ontology to integrate chemical, genomic and proteomic data with disease, treatment, and electronic health records. We demonstrate the use of Semantic Web technologies in the integration of patient and biomedical data, and reveal how such a knowledge base can aid physicians in providing tailored patient care and facilitate the recruitment of patients into active clinical trials. Thus, patients, physicians and researchers may explore the knowledge base to better understand therapeutic options, efficacy, and mechanisms of action. CONCLUSIONS: This work takes an important step in using Semantic Web technologies to facilitate integration of relevant, distributed, external sources and progress towards a computational platform to support personalized medicine. AVAILABILITY: TMO can be downloaded from http://code.google.com/p/translationalmedicineontology and TMKB can be accessed at http://tm.semanticscience.org/sparql. PMID- 21624156 TI - Integration and publication of heterogeneous text-mined relationships on the Semantic Web. AB - BACKGROUND: Advances in Natural Language Processing (NLP) techniques enable the extraction of fine-grained relationships mentioned in biomedical text. The variability and the complexity of natural language in expressing similar relationships causes the extracted relationships to be highly heterogeneous, which makes the construction of knowledge bases difficult and poses a challenge in using these for data mining or question answering. RESULTS: We report on the semi-automatic construction of the PHARE relationship ontology (the PHArmacogenomic RElationships Ontology) consisting of 200 curated relations from over 40,000 heterogeneous relationships extracted via text-mining. These heterogeneous relations are then mapped to the PHARE ontology using synonyms, entity descriptions and hierarchies of entities and roles. Once mapped, relationships can be normalized and compared using the structure of the ontology to identify relationships that have similar semantics but different syntax. We compare and contrast the manual procedure with a fully automated approach using WordNet to quantify the degree of integration enabled by iterative curation and refinement of the PHARE ontology. The result of such integration is a repository of normalized biomedical relationships, named PHARE-KB, which can be queried using Semantic Web technologies such as SPARQL and can be visualized in the form of a biological network. CONCLUSIONS: The PHARE ontology serves as a common semantic framework to integrate more than 40,000 relationships pertinent to pharmacogenomics. The PHARE ontology forms the foundation of a knowledge base named PHARE-KB. Once populated with relationships, PHARE-KB (i) can be visualized in the form of a biological network to guide human tasks such as database curation and (ii) can be queried programmatically to guide bioinformatics applications such as the prediction of molecular interactions. PHARE is available at http://purl.bioontology.org/ontology/PHARE. PMID- 21624157 TI - How orthogonal are the OBO Foundry ontologies? AB - BACKGROUND: Ontologies in biomedicine facilitate information integration, data exchange, search and query of biomedical data, and other critical knowledge intensive tasks. The OBO Foundry is a collaborative effort to establish a set of principles for ontology development with the eventual goal of creating a set of interoperable reference ontologies in the domain of biomedicine. One of the key requirements to achieve this goal is to ensure that ontology developers reuse term definitions that others have already created rather than create their own definitions, thereby making the ontologies orthogonal. METHODS: We used a simple lexical algorithm to analyze the extent to which the set of OBO Foundry candidate ontologies identified from September 2009 to September 2010 conforms to this vision. Specifically, we analyzed (1) the level of explicit term reuse in this set of ontologies, (2) the level of overlap, where two ontologies define similar terms independently, and (3) how the levels of reuse and overlap changed during the course of this year. RESULTS: We found that 30% of the ontologies reuse terms from other Foundry candidates and 96% of the candidate ontologies contain terms that overlap with terms from the other ontologies. We found that while term reuse increased among the ontologies between September 2009 and September 2010, the level of overlap among the ontologies remained relatively constant. Additionally, we analyzed the six ontologies announced as OBO Foundry members on March 5, 2010, and identified that the level of overlap was extremely low, but, notably, so was the level of term reuse. CONCLUSIONS: We have created a prototype web application that allows OBO Foundry ontology developers to see which classes from their ontologies overlap with classes from other ontologies in the OBO Foundry (http://obomap.bioontology.org). From our analysis, we conclude that while the OBO Foundry has made significant progress toward orthogonality during the period of this study through increased adoption of explicit term reuse, a large amount of overlap remains among these ontologies. Furthermore, the characteristics of the identified overlap, such as the terms it comprises and its distribution among the ontologies, indicate that the achieving orthogonality will be exceptionally difficult, if not impossible. PMID- 21624158 TI - HyQue: evaluating hypotheses using Semantic Web technologies. AB - BACKGROUND: Key to the success of e-Science is the ability to computationally evaluate expert-composed hypotheses for validity against experimental data. Researchers face the challenge of collecting, evaluating and integrating large amounts of diverse information to compose and evaluate a hypothesis. Confronted with rapidly accumulating data, researchers currently do not have the software tools to undertake the required information integration tasks. RESULTS: We present HyQue, a Semantic Web tool for querying scientific knowledge bases with the purpose of evaluating user submitted hypotheses. HyQue features a knowledge model to accommodate diverse hypotheses structured as events and represented using Semantic Web languages (RDF/OWL). Hypothesis validity is evaluated against experimental and literature-sourced evidence through a combination of SPARQL queries and evaluation rules. Inference over OWL ontologies (for type specifications, subclass assertions and parthood relations) and retrieval of facts stored as Bio2RDF linked data provide support for a given hypothesis. We evaluate hypotheses of varying levels of detail about the genetic network controlling galactose metabolism in Saccharomyces cerevisiae to demonstrate the feasibility of deploying such semantic computing tools over a growing body of structured knowledge in Bio2RDF. CONCLUSIONS: HyQue is a query-based hypothesis evaluation system that can currently evaluate hypotheses about the galactose metabolism in S. cerevisiae. Hypotheses as well as the supporting or refuting data are represented in RDF and directly linked to one another allowing scientists to browse from data to hypothesis and vice versa. HyQue hypotheses and data are available at http://semanticscience.org/projects/hyque. PMID- 21624159 TI - An open annotation ontology for science on web 3.0. AB - BACKGROUND: There is currently a gap between the rich and expressive collection of published biomedical ontologies, and the natural language expression of biomedical papers consumed on a daily basis by scientific researchers. The purpose of this paper is to provide an open, shareable structure for dynamic integration of biomedical domain ontologies with the scientific document, in the form of an Annotation Ontology (AO), thus closing this gap and enabling application of formal biomedical ontologies directly to the literature as it emerges. METHODS: Initial requirements for AO were elicited by analysis of integration needs between biomedical web communities, and of needs for representing and integrating results of biomedical text mining. Analysis of strengths and weaknesses of previous efforts in this area was also performed. A series of increasingly refined annotation tools were then developed along with a metadata model in OWL, and deployed for feedback and additional requirements the ontology to users at a major pharmaceutical company and a major academic center. Further requirements and critiques of the model were also elicited through discussions with many colleagues and incorporated into the work. RESULTS: This paper presents Annotation Ontology (AO), an open ontology in OWL-DL for annotating scientific documents on the web. AO supports both human and algorithmic content annotation. It enables "stand-off" or independent metadata anchored to specific positions in a web document by any one of several methods. In AO, the document may be annotated but is not required to be under update control of the annotator. AO contains a provenance model to support versioning, and a set model for specifying groups and containers of annotation. AO is freely available under open source license at http://purl.org/ao/, and extensive documentation including screencasts is available on AO's Google Code page: http://code.google.com/p/annotation-ontology/ . CONCLUSIONS: The Annotation Ontology meets critical requirements for an open, freely shareable model in OWL, of annotation metadata created against scientific documents on the Web. We believe AO can become a very useful common model for annotation metadata on Web documents, and will enable biomedical domain ontologies to be used quite widely to annotate the scientific literature. Potential collaborators and those with new relevant use cases are invited to contact the authors. PMID- 21624160 TI - Automating generation of textual class definitions from OWL to English. AB - BACKGROUND: Text definitions for entities within bio-ontologies are a cornerstone of the effort to gain a consensus in understanding and usage of those ontologies. Writing these definitions is, however, a considerable effort and there is often a lag between specification of the main part of an ontology (logical descriptions and definitions of entities) and the development of the text-based definitions. The goal of natural language generation (NLG) from ontologies is to take the logical description of entities and generate fluent natural language. The application described here uses NLG to automatically provide text-based definitions from an ontology that has logical descriptions of its entities, so avoiding the bottleneck of authoring these definitions by hand. RESULTS: To produce the descriptions, the program collects all the axioms relating to a given entity, groups them according to common structure, realises each group through an English sentence, and assembles the resulting sentences into a paragraph, to form as 'coherent' a text as possible without human intervention. Sentence generation is accomplished using a generic grammar based on logical patterns in OWL, together with a lexicon for realising atomic entities. We have tested our output for the Experimental Factor Ontology (EFO) using a simple survey strategy to explore the fluency of the generated text and how well it conveys the underlying axiomatisation. Two rounds of survey and improvement show that overall the generated English definitions are found to convey the intended meaning of the axiomatisation in a satisfactory manner. The surveys also suggested that one form of generated English will not be universally liked; that intrusion of too much 'formal ontology' was not liked; and that too much explicit exposure of OWL semantics was also not liked. CONCLUSIONS: Our prototype tools can generate reasonable paragraphs of English text that can act as definitions. The definitions were found acceptable by our survey and, as a result, the developers of EFO are sufficiently satisfied with the output that the generated definitions have been incorporated into EFO. Whilst not a substitute for hand-written textual definitions, our generated definitions are a useful starting point. AVAILABILITY: An on-line version of the NLG text definition tool can be found at http://swat.open.ac.uk/tools/. The questionaire and sample generated text definitions may be found at http://mcs.open.ac.uk/nlg/SWAT/bio-ontologies.html. PMID- 21624161 TI - Scalable representations of diseases in biomedical ontologies. AB - BACKGROUND: The realm of pathological entities can be subdivided into pathological dispositions, pathological processes, and pathological structures. The latter are the bearer of dispositions, which can then be realized by their manifestations - pathologic processes. Despite its ontological soundness, implementing this model via purpose-oriented domain ontologies will likely require considerable effort, both in ontology construction and maintenance, which constitutes a considerable problem for SNOMED CT, presently the largest biomedical ontology. RESULTS: We describe an ontology design pattern which allows ontologists to make assertions that blur the distinctions between dispositions, processes, and structures until necessary. Based on the domain upper-level ontology BioTop, it permits ascriptions of location and participation in the definition of pathological phenomena even without an ontological commitment to a distinction between these three categories. An analysis of SNOMED CT revealed that numerous classes in the findings/disease hierarchy are ambiguous with respect to process vs. disposition. Here our proposed approach can easily be applied to create unambiguous classes. No ambiguities could be defined regarding the distinction of structure and non-structure classes, but here we have found problematic duplications. CONCLUSIONS: We defend a judicious use of disjunctive, and therefore ambiguous, classes in biomedical ontologies during the process of ontology construction and in the practice of ontology application. The use of these classes is permitted to span across several top-level categories, provided it contributes to ontology simplification and supports the intended reasoning scenarios. PMID- 21624162 TI - Developing a kidney and urinary pathway knowledge base. AB - BACKGROUND: Chronic renal disease is a global health problem. The identification of suitable biomarkers could facilitate early detection and diagnosis and allow better understanding of the underlying pathology. One of the challenges in meeting this goal is the necessary integration of experimental results from multiple biological levels for further analysis by data mining. Data integration in the life science is still a struggle, and many groups are looking to the benefits promised by the Semantic Web for data integration. RESULTS: We present a Semantic Web approach to developing a knowledge base that integrates data from high-throughput experiments on kidney and urine. A specialised KUP ontology is used to tie the various layers together, whilst background knowledge from external databases is incorporated by conversion into RDF. Using SPARQL as a query mechanism, we are able to query for proteins expressed in urine and place these back into the context of genes expressed in regions of the kidney. CONCLUSIONS: The KUPKB gives KUP biologists the means to ask queries across many resources in order to aggregate knowledge that is necessary for answering biological questions. The Semantic Web technologies we use, together with the background knowledge from the domain's ontologies, allows both rapid conversion and integration of this knowledge base. The KUPKB is still relatively small, but questions remain about scalability, maintenance and availability of the knowledge itself. AVAILABILITY: The KUPKB may be accessed via http://www.e-lico.eu/kupkb. PMID- 21624163 TI - Mining of vaccine-associated IFN-gamma gene interaction networks using the Vaccine Ontology. AB - BACKGROUND: Interferon-gamma (IFN-gamma) is vital in vaccine-induced immune defense against bacterial and viral infections and tumor. Our recent study demonstrated the power of a literature-based discovery method in extraction and comparison of the IFN-gamma and vaccine-mediated gene interaction networks. The Vaccine Ontology (VO) contains a hierarchy of vaccine names. It is hypothesized that the application of VO will enhance the prediction of IFN-gamma and vaccine mediated gene interaction network. RESULTS: In this study, 186 specific vaccine names listed in the Vaccine Ontology (VO) and their semantic relations were used for possible improved retrieval of the IFN-gamma and vaccine associated gene interactions. The application of VO allows discovery of 38 more genes and 60 more interactions. Comparison of different layers of IFN-gamma networks and the example BCG vaccine-induced subnetwork led to generation of new hypotheses. By analyzing all discovered genes using centrality metrics, 32 genes were ranked high in the VO-based IFN-gamma vaccine network using four centrality scores. Furthermore, 28 specific vaccines were found to be associated with these top 32 genes. These specific vaccine-gene associations were further used to generate a network of vaccine-vaccine associations. The BCG and LVS vaccines are found to be the most central vaccines in the vaccine-vaccine association network. CONCLUSION: Our results demonstrate that the combined usages of biomedical ontologies and centrality-based literature mining are able to significantly facilitate discovery of gene interaction networks and gene-concept associations. AVAILABILITY: VO is available at: http://www.violinet.org/vaccineontology; and the SVM edit kernel for gene interaction extraction is available at: http://www.violinet.org/ifngvonet/int_ext_svm.zip. PMID- 21624164 TI - Representation of research hypotheses. AB - BACKGROUND: Hypotheses are now being automatically produced on an industrial scale by computers in biology, e.g. the annotation of a genome is essentially a large set of hypotheses generated by sequence similarity programs; and robot scientists enable the full automation of a scientific investigation, including generation and testing of research hypotheses. RESULTS: This paper proposes a logically defined way for recording automatically generated hypotheses in machine amenable way. The proposed formalism allows the description of complete hypotheses sets as specified input and output for scientific investigations. The formalism supports the decomposition of research hypotheses into more specialised hypotheses if that is required by an application. Hypotheses are represented in an operational way - it is possible to design an experiment to test them. The explicit formal description of research hypotheses promotes the explicit formal description of the results and conclusions of an investigation. The paper also proposes a framework for automated hypotheses generation. We demonstrate how the key components of the proposed framework are implemented in the Robot Scientist "Adam". CONCLUSIONS: A formal representation of automatically generated research hypotheses can help to improve the way humans produce, record, and validate research hypotheses. AVAILABILITY: http://www.aber.ac.uk/en/cs/research/cb/projects/robotscientist/results/ PMID- 21624165 TI - Comparison of analyses of the QTLMAS XIV common dataset. I: genomic selection. AB - BACKGROUND: For the XIV QTLMAS workshop, a dataset for traits with complex genetic architecture has been simulated and released for analyses by participants. One of the tasks was to estimate direct genomic values for individuals without phenotypes. The aim of this paper was to compare results of different approaches used by the participants to calculate direct genomic values for quantitative trait (QT) and binary trait (BT). RESULTS: Participants applied 26 approaches for QT and 15 approaches for BT. Accuracy for QT was between 0.26 and 0.89 for males and between 0.31 and 0.89 for females, and for BT ranged from 0.27 to 0.85. For QT, percentage of lost response to selection varied from 8% to 83%, whereas for BT the loss was between 15% and 71%. CONCLUSIONS: Bayesian model averaging methods predicted breeding values slightly better than GBLUP in a simulated data set. The methods utilizing genomic information performed better than traditional pedigree based BLUP analyses. Bivariate analyses was slightly advantageous over single trait for the same method. None of the methods estimated the non-additivity of QTL affecting the QT, which may be one of the constrains in accuracy observed in real data. PMID- 21624166 TI - Haplotype inference based on Hidden Markov Models in the QTL-MAS 2010 multi generational dataset. AB - BACKGROUND: We have previously demonstrated an approach for efficient computation of genotype probabilities, and more generally probabilities of allele inheritance in inbred as well as outbred populations. That work also included an extension for haplotype inference, or phasing, using Hidden Markov Models. Computational phasing of multi-thousand marker datasets has not become common as of yet. In this communication, we further investigate the method presented earlier for such problems, in a multi-generational dataset simulated for QTL detection. RESULTS: When analyzing the dataset simulated for the 14th QTLMAS workshop, the phasing produced showed zero deviations compared to original simulated phase in the founder generation. In total, 99.93% of all markers were correctly phased. 97.68% of the individuals were correct in all markers over all 5 simulated chromosomes. Results were produced over a weekend on a small computational cluster. The specific algorithmic adaptations needed for the Markov model training approach in order to reach convergence are described. CONCLUSIONS: Our method provides efficient, near-perfect haplotype inference allowing the determination of completely phased genomes in dense pedigrees. These developments are of special value for applications where marker alleles are not corresponding directly to QTL alleles, thus necessitating tracking of allele origin, and in complex multi generational crosses. The cnF2freq codebase, which is in a current state of active development, is available under a BSD-style license. PMID- 21624167 TI - A comparison of random forests, boosting and support vector machines for genomic selection. AB - BACKGROUND: Genomic selection (GS) involves estimating breeding values using molecular markers spanning the entire genome. Accurate prediction of genomic breeding values (GEBVs) presents a central challenge to contemporary plant and animal breeders. The existence of a wide array of marker-based approaches for predicting breeding values makes it essential to evaluate and compare their relative predictive performances to identify approaches able to accurately predict breeding values. We evaluated the predictive accuracy of random forests (RF), stochastic gradient boosting (boosting) and support vector machines (SVMs) for predicting genomic breeding values using dense SNP markers and explored the utility of RF for ranking the predictive importance of markers for pre-screening markers or discovering chromosomal locations of QTLs. METHODS: We predicted GEBVs for one quantitative trait in a dataset simulated for the QTLMAS 2010 workshop. Predictive accuracy was measured as the Pearson correlation between GEBVs and observed values using 5-fold cross-validation and between predicted and true breeding values. The importance of each marker was ranked using RF and plotted against the position of the marker and associated QTLs on one of five simulated chromosomes. RESULTS: The correlations between the predicted and true breeding values were 0.547 for boosting, 0.497 for SVMs, and 0.483 for RF, indicating better performance for boosting than for SVMs and RF. CONCLUSIONS: Accuracy was highest for boosting, intermediate for SVMs and lowest for RF but differed little among the three methods and relative to ridge regression BLUP (RR-BLUP). PMID- 21624168 TI - Pre-selection of markers for genomic selection. AB - BACKGROUND: Accurate prediction of genomic breeding values (GEBVs) requires numerous markers. However, predictive accuracy can be enhanced by excluding markers with no effects or with inconsistent effects among crosses that can adversely affect the prediction of GEBVs. METHODS: We present three different approaches for pre-selecting markers prior to predicting GEBVs using four different BLUP methods, including ridge regression and three spatial models. Performances of the models were evaluated using 5-fold cross-validation. RESULTS AND CONCLUSIONS: Ridge regression and the spatial models gave essentially similar fits. Pre-selecting markers was evidently beneficial since excluding markers with inconsistent effects among crosses increased the correlation between GEBVs and true breeding values of the non-phenotyped individuals from 0.607 (using all markers) to 0.625 (using pre-selected markers). Moreover, extension of the ridge regression model to allow for heterogeneous variances between the most significant subset and the complementary subset of pre-selected markers increased predictive accuracy (from 0.625 to 0.648) for the simulated dataset for the QTL MAS 2010 workshop. PMID- 21624169 TI - Genomic breeding value prediction and QTL mapping of QTLMAS2010 data using Bayesian Methods. AB - BACKGROUND: Bayesian methods allow prediction of genomic breeding values (GEBVs) using high-density single nucleotide polymorphisms (SNPs) covering the whole genome with effective shrinkage of SNP effects using appropriate priors. In this study we applied a modification of the well-known BayesA and BayesB methods to estimate the proportion of SNPs with zero effects (pi) and a common variance for non-zero effects. The method, termed BayesCpi, was used to predict the GEBVs of the last generation of the QTLMAS2010 data. The accuracy of GEBVs from various methods was estimated by the correlation with phenotypes in the last generation. The methods were BayesCPi and BayesB with different pi values, both with and without polygenic effects, and best linear unbiased prediction using an animal model with a genomic or numerator relationship matrix. Positions of quantitative trait loci (QTLs) were identified based on the variances of GEBVs for windows of 10 consecutive SNPs. We also proposed a novel approach to set significance thresholds for claiming QTL in this specific case by using pedigree-based simulation of genotypes. All analyses were focused on detecting and evaluating QTL with additive effects. RESULTS: The accuracy of GEBVs was highest for BayesCpi, but the accuracy of BayesB with pi equal to 0.99 was similar to that of BayesCpi. The accuracy of BayesB dropped with a decrease in pi. Including polygenic effects into the model only had marginal effects on accuracy and bias of predictions. The number of QTL identified was 15 when based on a stringent 10% chromosome-wise threshold and increased to 21 when a 20% chromosome-wise threshold was used. CONCLUSIONS: The BayesCpi method without polygenic effects was identified to be the best method for the QTLMAS2010 dataset, because it had highest accuracy and least bias. The significance criterion based on variance of 10-SNP windows allowed detection of more than half of the QTL, with few false positives. PMID- 21624170 TI - Hierarchical likelihood opens a new way of estimating genetic values using genome wide dense marker maps. AB - BACKGROUND: Genome-wide dense markers have been used to detect genes and estimate relative genetic values. Among many methods, Bayesian techniques have been widely used and shown to be powerful in genome-wide breeding value estimation and association studies. However, computation is known to be intensive under the Bayesian framework, and specifying a prior distribution for each parameter is always required for Bayesian computation. We propose the use of hierarchical likelihood to solve such problems. RESULTS: Using double hierarchical generalized linear models, we analyzed the simulated dataset provided by the QTLMAS 2010 workshop. Marker-specific variances estimated by double hierarchical generalized linear models identified the QTL with large effects for both the quantitative and binary traits. The QTL positions were detected with very high accuracy. For young individuals without phenotypic records, the true and estimated breeding values had Pearson correlation of 0.60 for the quantitative trait and 0.72 for the binary trait, where the quantitative trait had a more complicated genetic architecture involving imprinting and epistatic QTL. CONCLUSIONS: Hierarchical likelihood enables estimation of marker-specific variances under the likelihoodist framework. Double hierarchical generalized linear models are powerful in localizing major QTL and computationally fast. PMID- 21624171 TI - Genomic selection for QTL-MAS data using a trait-specific relationship matrix. AB - BACKGROUND: The genomic estimated breeding values (GEBV) of the young individuals in the XIV QTL-MAS workshop dataset were predicted by three methods: best linear unbiased prediction with a trait-specific marker-derived relationship matrix (TABLUP), ridge regression best linear unbiased prediction (RRBLUP), and BayesB. METHODS: The TABLUP method is identical to the conventional BLUP except that the numeric relationship matrix is replaced with a trait-specific marker-derived relationship matrix (TA). The TA matrix was constructed based on both marker genotypes and their estimated effects on the trait of interest. The marker effects were estimated in a reference population consisting of 2 326 individuals using RRBLUP and BayesB. The GEBV of individuals in the reference population as well as 900 young individuals were estimated using the three methods. Subsets of markers were selected to perform low-density marker genomic selection for TABLUP method. RESULTS: The correlations between GEBVs from different methods are over 0.95 in most scenarios. The correlations between BayesB using all markers and TABLUP using 200 or more selected markers to construct the TA matrix are higher than 0.98 in the candidate population. The accuracy of TABLUP is higher than 0.67 with 100 or more selected markers, which is nearly equal to the accuracy of BayesB with all markers. CONCLUSIONS: TABLUP method performed nearly equally to BayesB method with the common dataset. It also provides an alternative method to predict GEBV with low-density markers. TABLUP is therefore a promising method for genomic selection deserving further exploration. PMID- 21624172 TI - Comparison of analyses of the QTLMAS XIV common dataset. II: QTL analysis. AB - BACKGROUND: A quantitative and a binary trait for the 14th QTLMAS 2010 workshop were simulated under a model which combined additive inheritance, epistasis and imprinting. This paper aimed to compare results submitted by the participants of the workshop. METHODS: The results were compared according to three criteria: the success rate (ratio of mapped QTL to the total number of simulated QTL), and the error rate (ratio of false positives to the number of reported positions), and mean distance between a true mapped QTL and the nearest submitted position. RESULTS: Seven groups submitted results for the quantitative trait and five for the binary trait. Among the 37 simulated QTL 17 remained undetected. Success rate ranged from 0.05 to 0.43, error rate was between 0.00 and 0.92, and the mean distance ranged from 0.26 to 0.77 Mb. CONCLUSIONS: Our comparison shows that differences among methods used by the participants increases with the complexity of genetic architecture. It was particularly visible for the quantitative trait which was determined partly by non-additive QTL. Furthermore, an imprinted QTL with a large effect may remain undetected if the applied model tests only for Mendelian genes. PMID- 21624173 TI - QTLMAS 2010: simulated dataset. AB - BACKGROUND: Objective was to simulate the data for the QTLMAS 2010 Workshop under a model that includes major additive, epistatic and parent-of-origin effects. RESULTS: Data were simulated for 3226 individuals in 5 generations. Genomic data for 5 chromosomes were simulated using coalescent model. In total, the data included 10,031 SNPs, 30 additive QTLs, 2 interacting QTL pairs, and 3 imprinted loci. The density was 20 SNPs/1Mb, whereas mean linkage disequilibrium between adjacent SNPs was 0.1. One quantitative and one binary trait were simulated with heritability of 0.39-0.52 and additive correlation of 0.59. The data can be used as a benchmark for comparison of QTL mapping methods and models for genomic breeding value estimation under complex genetic architecture. PMID- 21624174 TI - A Bayesian approach to detect QTL affecting a simulated binary and quantitative trait. AB - BACKGROUND: We analyzed simulated data from the 14th QTL-MAS workshop using a Bayesian approach implemented in the program iBay. The data contained individuals genotypes for 10,031 SNPs and phenotyped for a quantitative and a binary trait. RESULTS: For the quantitative trait we mapped 8 out of 30 additive QTL, 1 out of 3 imprinted QTL and both epistatic pairs of QTL successfully. For the binary trait we mapped 11 out of 22 additive QTL successfully. Four out of 22 pleiotropic QTL were detected as such. CONCLUSIONS: The Bayesian variable selection method showed to be a successful method for genome-wide association. This method was reasonably fast using dense marker maps. PMID- 21624175 TI - Estimating genomic breeding values and detecting QTL using univariate and bivariate models. AB - BACKGROUND: Genomic selection is particularly beneficial for difficult or expensive to measure traits. Since multi-trait selection is an important tool to deal with such cases, an important question is what the added value is of multi trait genomic selection. METHODS: The simulated dataset, including a quantitative and binary trait, was analyzed with four univariate and bivariate linear models to predict breeding values for juvenile animals. Two models estimated variance components with REML using a numerator (A), or SNP based relationship matrix (G). Two SNP based Bayesian models included one (BayesA) or two distributions (BayesC) for estimated SNP effects. The bivariate BayesC model sampled QTL probabilities for each SNP conditional on both traits. Genotypes were permuted 2,000 times against phenotypes and pedigree, to obtain significance thresholds for posterior QTL probabilities. Genotypes were permuted rather than phenotypes, to retain relationships between pedigree and phenotypes, such that polygenic effects could still be estimated. RESULTS: Correlations between estimated breeding values (EBV) of different SNP based models, for juvenile animals, were greater than 0.93 (0.87) for the quantitative (binary) trait. Estimated genetic correlation was 0.71 (0.66) for model G (A). Accuracies of breeding values of SNP based models were for both traits highest for BayesC and lowest for G. Accuracies of breeding values of bivariate models were up to 0.08 higher than for univariate models.The bivariate BayesC model detected 14 out of 32 QTL for the quantitative trait, and 8 out of 22 for the binary trait. CONCLUSIONS: Accuracy of EBV clearly improved for both traits using bivariate compared to univariate models. BayesC achieved highest accuracies of EBV and was also one of the methods that found most QTL. Permuting genotypes against phenotypes and pedigree in BayesC provided an effective way to derive significance thresholds for posterior QTL probabilities. PMID- 21624176 TI - Genotype imputation for the prediction of genomic breeding values in non genotyped and low-density genotyped individuals. AB - BACKGROUND: There is wide interest in calculating genomic breeding values (GEBVs) in livestock using dense, genome-wide SNP data. The general framework for genomic selection assumes all individuals are genotyped at high-density, which may not be true in practice. Methods to add additional genotypes for individuals not genotyped at high density have the potential to increase GEBV accuracy with little or no additional cost. In this study a long haplotype library was created using a long range phasing algorithm and used in combination with segregation analysis to impute dense genotypes for non-genotyped dams in the training dataset (S1) and for non-genotyped or low-density genotyped individuals in the prediction dataset (S2), using the 14th QTL-MAS Workshop dataset. Alternative low-density scenarios were evaluated for accuracy of imputed genotypes and prediction of GEBVs. RESULTS: In S1, females in the training population were not genotyped and prediction individuals were either not genotyped or genotyped at low-density (evenly spaced at 2, 5 or 10 Mb). The proportion of correctly imputed genotypes for training females did not change when genotypes were added for individuals in the prediction set whereas the number of correctly imputed genotypes in the prediction set increased slightly (S1). The S2 scenario assumed the complete training set was genotyped for all SNPs and the prediction set was not genotyped or genotyped at low-density. The number of correctly imputed genotypes increased with genotyping density in the prediction set. Accuracy of genomic breeding values for the prediction set in each scenario were the correlation of GEBVs with true breeding values and were used to evaluate the potential loss in accuracy with reduced genotyping. For both S1 and S2 the GEBV accuracies were similar when the prediction set was not genotyped and increased with the addition of low density genotypes, with the increase larger for S2 than S1. CONCLUSIONS: Genotype imputation using a long haplotype library and segregation analysis is promising for application in sparsely-genotyped pedigrees. The results of this study suggest that dense genotypes can be imputed for selection candidates with some loss in genomic breeding value accuracy, but with levels of accuracy higher than traditional BLUP estimated breeding values. Accurate genotype imputation would allow for a single low-density SNP panel to be used across traits. PMID- 21624177 TI - Partial least square regression applied to the QTLMAS 2010 dataset. AB - BACKGROUND: Partial least square regression (PLSR) was used to analyze the data of the QTLMAS 2010 workshop to identify genomic regions affecting either one of the two traits and to estimate breeding values. PLSR was appropriate for these data because it enabled to simultaneously fit several traits to the markers. RESULTS: A preliminary analysis showed phenotypic and genetic correlations between the two traits. Consequently, the data were analyzed jointly in a PLSR model for each chromosome independently. Regression coefficients for the markers were used to calculate the variance of each marker and inference of quantitative trait loci (QTL) was based on local maxima of a smoothed line traced through these variances. In this way, 25 QTL for the continuous trait and 22 for the discrete trait were found. There was evidence for pleiotropic QTL on chromosome 1. The 2000 most important markers were fitted in a second PLSR model to calculate breeding values of the individuals. The accuracies of these estimated breeding values ranged between 0.56 and 0.92. CONCLUSIONS: Results showed the viability of PLSR for QTL analysis and estimating breeding values using markers. PMID- 21624178 TI - Association analyses of the MAS-QTL data set using grammar, principal components and Bayesian network methodologies. AB - BACKGROUND: It has been shown that if genetic relationships among individuals are not taken into account for genome wide association studies, this may lead to false positives. To address this problem, we used Genome-wide Rapid Association using Mixed Model and Regression and principal component stratification analyses. To account for linkage disequilibrium among the significant markers, principal components loadings obtained from top markers can be included as covariates. Estimation of Bayesian networks may also be useful to investigate linkage disequilibrium among SNPs and their relation with environmental variables.For the quantitative trait we first estimated residuals while taking polygenic effects into account. We then used a single SNP approach to detect the most significant SNPs based on the residuals and applied principal component regression to take linkage disequilibrium among these SNPs into account. For the categorical trait we used principal component stratification methodology to account for background effects. For correction of linkage disequilibrium we used principal component logit regression. Bayesian networks were estimated to investigate relationship among SNPs. RESULTS: Using the Genome-wide Rapid Association using Mixed Model and Regression and principal component stratification approach we detected around 100 significant SNPs for the quantitative trait (p<0.05 with 1000 permutations) and 109 significant (p<0.0006 with local FDR correction) SNPs for the categorical trait. With additional principal component regression we reduced the list to 16 and 50 SNPs for the quantitative and categorical trait, respectively. CONCLUSIONS: GRAMMAR could efficiently incorporate the information regarding random genetic effects. Principal component stratification should be cautiously used with stringent multiple hypothesis testing correction to correct for ancestral stratification and association analyses for binary traits when there are systematic genetic effects such as half sib family structures. Bayesian networks are useful to investigate relationships among SNPs and environmental variables. PMID- 21624179 TI - Applying different genomic evaluation approaches on QTLMAS2010 dataset. AB - BACKGROUND: With the availability of high throughput genotyping, genomic selection, the evaluation of animals based on dense SNP genotyping, is receiving more and more attention. Several statistical methods have been suggested for genomic selection. Compared to traditional selection, genomic selection can be more accurate which can lead to higher efficiency in terms of time and cost. Herein we applied different genomic evaluation methods on the 14th QTLMAS dataset. METHODS: Four different approaches were used for the estimation of EBV of animals for the Quantitative and the Binary Trait (QT and BT respectively). It included two Bayes B types of approaches (BB): using only SNP information (GBB) or SNP and Pedigree information (GPBB); and two genomic BLUP, GBLUP and GPBLUP. Traditional BLUP was also used only for comparison. When using BB methodology, the probability of SNP having an effect on the traits (which include a quantitative and a binary trait) were also estimated. We also performed "standard" QTL mapping approaches including linkage and association analyses to compare them with BB results as a potential QTL mapping tools. RESULTS: For QT, the best accuracy of EBV (correlation between EBVs and TBVs) for young animals, was obtained by BB methods (r = 0.68). Genomic BLUP estimations (GBLUP and GPBLUP) were less accurate (r = 0.60 and 0.61 respectively). Similar results were obtained for the BT: r were estimated at 0.82, 0.82, 0.71 and 0.70 for GPBB, GBB, GPBLUP and GBLUP respectively. Using traditional BLUP, r was at 0.39 and 0.47 for QT and BT respectively. The genetic correlation between the two traits (approximated by the correlation between EBVs for BT and QT using GBB method) was as high as 0.58. CONCLUSIONS: Better accuracies were obtained using BB methods, compared to BLUP analyses. Compared to the traditional BLUP, the accuracy of the EBVs was improved about 70% and 50% using BB and GBLUP methods respectively. The benefit of genomic selection was the same for both the QT and BT. Models with and without polygenic effect led to similar accuracies in the estimation of breeding values. The BT and QT were genetically correlated (r=0.58) which suggested that bivariate analyses may be of advantages. Signal profile by GBB followed well the true QTL patterns, which was consistent with good estimation of EBVs by this method, suggesting its potential value for QTL mapping. PMID- 21624180 TI - Anti-TNF therapy: what have we learned in 12 years? PMID- 21624181 TI - Infliximab: 12 years of experience. AB - Rheumatoid arthritis (RA), ankylosing spondylitis (AS) and psoriatic arthritis (PsA) are immune-mediated conditions that share an inflammatory mechanism fuelled by excessive cytokines, particularly TNF. Control of inflammation and rapid suppression of cytokines are important in treating these diseases. With this understanding and the corresponding advent of TNF inhibitors, RA patients, AS patients and PsA patients have found more choices than ever before and have greater hope of sustained relief. As a widely used TNF inhibitor, infliximab has a deep and established record of efficacy and safety data. Extensive evidence - from randomised controlled clinical trials, large registries and postmarketing surveillance studies - shows that infliximab effectively treats the signs and symptoms, provides rapid and prolonged suppression of inflammation, prevents radiologically observable disease progression and offers an acceptable safety profile in RA, AS and PsA. In very recent studies, investigators have observed drug-free remission in some patients. Additionally, infliximab may interfere with rapidly progressing disease in RA by early addition to methotrexate in patients with signs of an aggressive course. Finally, infliximab has been shown to reduce PsA clinical manifestations such as nail involvement. With our current understanding, substantial data and increasing confidence regarding use in practice, infliximab can be considered a well-known drug in our continued campaign against inflammatory rheumatic diseases. PMID- 21624182 TI - Understanding emerging treatment paradigms in rheumatoid arthritis. AB - Treatment strategies for rheumatoid arthritis (RA) will continue to evolve as new drugs are developed, as new data become available, and as our potential to achieve greater and more consistent outcomes becomes more routine. Many patients will find both symptom relief and modest control of their disease with disease modifying antirheumatic drugs (DMARDs), yet this course of therapy is clearly not effective in all patients. In fact, despite strong evidence that intensive treatment in the early stages of RA can slow or stop disease progression and may prevent disability, many patients continue to be managed in a stepwise manner and are treated with an ongoing monotherapy regimen with DMARDs. There is now a large body of evidence demonstrating the success of treating RA patients with anti-TNF therapy, usually in combination with methotrexate. As a result of the increased use of anti-TNF therapy, treatment paradigms have changed - and our practice is beginning to reflect this change. In the present review, we summarize the salient points of several recently proposed and emerging treatment paradigms with an emphasis on how these strategies may impact future practice. PMID- 21624183 TI - Structural damage in rheumatoid arthritis, psoriatic arthritis, and ankylosing spondylitis: traditional views, novel insights gained from TNF blockade, and concepts for the future. AB - Structural changes of bone and cartilage are a hallmark of inflammatory joint diseases such as rheumatoid arthritis (RA), psoriatic arthritis (PsA), and ankylosing spondylitis (AS). Despite certain similarities - in particular, inflammation as the driving force for structural changes - the three major inflammatory joint diseases show considerably different pathologies. Whereas RA primarily results in bone and cartilage resorption, PsA combines destructive elements with anabolic bone responses, and AS is the prototype of a hyper responsive joint disease associated with substantial bone and cartilage apposition. In the present review we summarize the clinical picture and pathophysiologic processes of bone and cartilage damage in RA, PsA, and AS, we describe the key insights obtained from the introduction of TNF blockade, and we discuss the future challenges and frontiers of structural damage in arthritis. PMID- 21624185 TI - UN convention on the rights of persons with disability, eligibility criteria and the International Classification of Functioning Disability and Health. PMID- 21624184 TI - Advances in rheumatology: new targeted therapeutics. AB - Treatment of inflammatory arthritides - including rheumatoid arthritis, ankylosing spondylitis, and psoriatic arthritis - has seen much progress in recent years, partially due to increased understanding of the pathogenesis of these diseases at the cellular and molecular levels. These conditions share some common mechanisms. Biologic therapies have provided a clear advance in the treatment of rheumatological conditions. Currently available TNF-targeting biologic agents that are licensed for at east one of the above-named diseases are etanercept, infliximab, adalimumab, golimumab, and certolizumab. Biologic agents with a different mechanism of action have also been approved in rheumatoid arthritis (rituximab, abatacept, and tocilizumab). Although these biologic agents are highly effective, there is a need for improved management strategies. There is also a need for education of family physicians and other healthcare professionals in the identification of early symptoms of inflammatory arthritides and the importance of early referral to rheumatologists for diagnosis and treatment. Also, researchers are developing molecules - for example, the Janus kinase inhibitor CP-690550 (tofacitinib) and the spleen tyrosine kinase inhibitor R788 (fostamatinib) - to target other aspects of the inflammatory cascade. Initial trial results with new agents are promising, and, in time, head-to-head trials will establish the best treatment options for patients. The key challenge is identifying how best to integrate these new, advanced therapies into daily practice. PMID- 21624186 TI - "Evaluating the model of classification and valuation of disabilities used in Brazil and defining the elaboration and adoption of a unique model for all the country": Brazilian Interministerial Workgroup Task. AB - The President of Brazil established an Interministerial Work Group in order to "evaluate the model of classification and valuation of disabilities used in Brazil and to define the elaboration and adoption of a unique model for all the country". Eight Ministries and/or Secretaries participated in the discussion over a period of 10 months, concluding that a proposed model should be based on the United Nations Convention on the Rights of Person with Disabilities, the International Classification of Functioning, Disability and Health, and the 'support theory', and organizing a list of recommendations and necessary actions for a Classification, Evaluation and Certification Network with national coverage. PMID- 21624187 TI - Description of the person-environment interaction: methodological issues and empirical results of an Italian large-scale disability assessment study using an ICF-based protocol. AB - BACKGROUND: There is a connection between the definition of disability in a person-environment framework, the development of appropriate assessment strategies and instruments, and the logic underpinning the organization of benefits and services to confront disability. METHODS: The Italian Ministry of Health and Ministry of Labor and Social Policies supported a three-year project for the definition of a common framework and a standardised protocol for disability evaluation based on ICF. The research agenda of the project identified 6 phases: 1) adoption of a definition of disability; 2) analytical breakdown of the contents of disability definition, so as to indicate as clearly as possible the core information essential to guide the evaluation process; 3) definition of a data collection protocol; 4) national implementation of the protocol and collection of approximately 1,000 profiles; 5) proposal of a profile analysis and definition of groups of cases with similar functioning profiles; 6) trial of the proposal with the collected data. The data was analyzed in different ways: descriptive analysis, application of the person-environment interactions classification tree, and cluster analysis. RESULTS: A sample of 1,051 persons from 8 Italian regions was collected that represented different functioning conditions in all the phases of the life cycle. The aggregate result of the person-environment interactions was summarized. The majority of activities resulted with no problems in all of the A&P chapters. Nearly 50.000 facilitators codes were opened. The main frequent facilitators were family members, health and social professionals, assistive devices and both health and social systems, services and politics. The focus of the person-environment interaction evaluation was on the A&P domains, differentiating those in which performance presented limitations and restrictions from those in which performance had no or light limitations and restrictions. Communication(d3) and Learning and Applying Knowledge(d1) appeared as the more problematic A&P areas. Self Care(d5) was the domain in which facilitators were more effective in supporting functioning, suggesting that the Italian welfare system is mainly focused on providing care services for activities of daily living, jointly with the family. The cluster analysis was limited to those categories that were common to all age classes (38 categories out of 55). For a final representation, a solution with 6 clusters was chosen. CONCLUSIONS: An example is provided of how it is possible to plan empirical studies in which theoretical advances and operative goals on disability in a person-environment framework can support the definition of a research design, measurement strategies, and data analysis. The description of functioning and disability at population level is no more based on individual deficits or limitations. Personal profiles may be elaborated and groups created based on the characteristics of the person-environment interactions. Personal profiles may also be used as a "rationale" for defining personalized intervention programs. PMID- 21624188 TI - From codes to language: is the ICF a classification system or a dictionary? PMID- 21624189 TI - Use of The International Classification of Functioning, Disability and Health (ICF) as a conceptual framework and common language for disability statistics and health information systems. AB - A common framework for describing functional status information is needed in order to make this information comparable and of value. The World Health Organization's International Classification of Functioning, Disability and Health (ICF), which has been approved by all its member states, provides this common language and framework. The article provides an overview of ICF taxonomy, introduces the conceptual model which underpins ICF and elaborates on how ICF is used at population and clinical level. Furthermore, the article presents key features of the ICF tooling environment and outlines current and future developments of the classification. PMID- 21624190 TI - Measuring disability and monitoring the UN Convention on the Rights of Persons with Disabilities: the work of the Washington Group on Disability Statistics. AB - The Washington Group on Disability Statistics is a voluntary working group made up of representatives of over 100 National Statistical Offices and international, non-governmental and disability organizations that was organized under the aegis of the United Nations Statistical Division. The purpose of the Washington Group is to deal with the challenge of disability definition and measurement in a way that is culturally neutral and reasonably standardized among the UN member states. The work, which began in 2001, took on added importance with the passage and ratification of the UN Convention on the Rights of Persons with Disabilities since the Convention includes a provision for monitoring whether those with and without disabilities have equal opportunities to participate in society and this will require the identification of persons with disabilities in each nation. The International Classification of Functioning, Disability and Health (ICF) developed by the World Health Organization provided a framework for conceptualizing disability. Operationalizing an ICF-based approach to disability has required the development of new measurement tools for use in both censuses and surveys. To date, a short set of six disability-related questions suitable for use in national censuses has been developed and adopted by the Washington Group and incorporated by the United Nations in their Principles and Recommendations for Population and Housing Censuses. A series of extended sets of questions is currently under development and some of the sets have been tested in several countries. The assistance of many National and International organizations has allowed for cognitive and field testing of the disability questionnaires in multiple languages and locations. This paper will describe the work of the Washington Group and explicate the applicability of its approach and the questions developed for monitoring the UN Convention on the Rights of Persons with Disabilities. PMID- 21624191 TI - Using the ICF in Ireland. AB - This paper reflects on the use of ICF in Ireland, taking as a case study the experience of the first National Disability Survey (NDS). There were four clear effects in Ireland of using ICF as a framework for the NDS: a) that a broader range of people with disabilities was encompassed; b) that the environmental factors included from the ICF were comprehensive and policy relevant; c) that both barriers and facilitators were incorporated into the model; and d) that a focus on research ethics was encouraged. Some general conclusions regarding the benefits and limitations of ICF based on this experience are also drawn. PMID- 21624192 TI - Eligibility, the ICF and the UN Convention: Australian perspectives. AB - The UN Convention on the Rights of Persons with Disabilities, in Australia, acts as a philosophical and moral statement and framework guiding integrated and strategic policy across the nation. Broad policy agreement has been reached by governments, and both the government and non-government sectors are developing strategies for implementation or evaluation. There is however a need for a more integrated approach to disability policy and information, reflecting all three components of the Italian project: legislation and a high level philosophical framework and policy guide; a technical framework that can underpin specific policies and programs aiming to achieve the major goals; and , a language and set of tools, relating to both the above, that provide infrastructure for assessment methods and information systems. The International Classification of Functioning, Disability and Health (ICF) is the ideal tool to support the latter two components, consistent with the UN Convention. While the ICF has been used as the basis for national data standards, in population surveys and in the national data collection on disability support services, there is considerable scope for greater use of it, including using all domains of the Activities and Participation and the Environmental Factors component for policy, information and service provision, to advance a disability-inclusive society. Information available from the income support system and from generic services could be enhanced by reference to the ICF components. It would be of significant national value in Australia, especially as a 'continuum of care' is desired, if consistency of concepts and information were expanded across health and social welfare sectors. It would then be possible to obtain consistent data from health, aged care, disability and community services systems about key aspects of health and functioning, building a consolidated picture of access and experience across these sectors. Without attention to all three components of the Italian project and continuing effort to meet the challenges identified in this paper, it will not be possible to determine whether the goals of Australia's National Disability Agreement or the ambitions of the Convention are achieved. PMID- 21624193 TI - Development of an ICF-based eligibility procedure for education in Switzerland. AB - Starting in January 2011, Switzerland will implement a multidimensional, context sensitive procedure to establish eligibility in education systems. This paper provides a brief overview of the different eligibility-related practices with a special focus on children with disabilities. The paper then outlines the philosophical and conceptual framework of the eligibility procedure based on the International Classification of Functioning, Disability and Health, and the UN Convention on the Rights of Persons with Disability. The different components and methodology applied to organise information in the process towards establishing eligibility are also presented. Finally, some observations are made regarding transparent and just applications of the eligibility procedure, and the implementation of this new eligibility procedure. PMID- 21624194 TI - Monitoring the United Nation's Convention on the Rights of Persons with Disabilities: data and the International Classification of Functioning, Disability and Health. AB - This paper approaches the general issue of the complex challenges in the relationship between those who generate data--researchers, scientists, and state statistical offices--and those who use data--researchers and policy-makers--in light of the more specific policy challenges created by the monitoring requirement of the United Nation's Convention on the Rights of Persons with Disabilities (CRPD: Article 33). International Conventions and Treaties standardly suffer from being persistently ineffectual primarily because of the absence of implementation mechanisms. The CRPD, by contrast, explicitly requires State Parties who have ratified it to institute data generation and monitoring mechanisms for its implementation. This paper argues that WHO's International Classification of Functioning, Disability and Health (ICF) can be brought into the service of the CRPD data generation and monitoring mandate, both in the shaping of relevant data streams and in the creation of relevant indicators, and concludes by reviewing the challenges that remain. PMID- 21624195 TI - The WHO construct of health-related functioning (HrF) and its implications for health policy. AB - BACKGROUND: The International Classification of Functioning (ICF) has acquired a central role in the WHO Family of International Classifications and it has been extensively adopted as the reference framework for health-related functioning (HrF). This review aims to provide a description of the ICF/HrF to contextualise ICF/HrF in relation to other approaches to health functioning and to describe its application in policy and legislation with a special focus on Spain. METHODS: Narrative review based on the scientific literature and prior expert knowledge. RESULTS: ICF is both a coding system and a conceptual framework of HrF, which is framed as a unidimensional, bipolar and asymmetric construct with a negative pole (disability) and a positive pole (good functioning) with higher complexity. Other models of HrF include health promotion, quality of life and activities of daily living (ADL). The curtailed taxonomy of ICF and its unclear distinction from other approaches have had significant implications for research, policy and legislation, as illustrated by the case of the legislation and services for functional dependency in Spain and other examples. CONCLUSIONS: The ICF model of functioning is more comprehensive and usable than previous alternatives, but a full taxonomy of the HrF construct is needed to avoid further confusions in this field. This should also comprise harmonization with other classifications of the WHO Family of International Classifications and other models of health functioning. PMID- 21624196 TI - [Pay attention to diagnostic strategies for inflammatory bowel diseases in children]. PMID- 21624197 TI - [Alerts from "super bug"]. PMID- 21624198 TI - [Effect of zinc deficiency on intestinal mucosal morphology and digestive enzyme activity in growing rat]. AB - OBJECTIVE: In this study, a growing rat model of zinc deficiency was established to investigate the effect of zinc deficiency on intestinal mucosal morphology and digestive enzyme activity as well as to provide a scientific basis for zinc supplementation therapy in patients with diarrhea. METHOD: Three-week-old weaned Sprague-Dawley male rats (n = 30) were randomly divided into 3 groups with 10 in each: rats in the control group (ZA) were fed with a normal diet containing 30 ug/g zinc; rats in the zinc deficient group (ZD) were fed with a zinc-deficient diet containing 0.4 ug/g zinc (refer to AIN-76 formula); and rats in the paired fed group (PF) were fed with a normal diet, but the food intake was limited to intake of rats in ZD group in the previous day. All rats were provided with deionized water for drinking. Their body weight was measured and the food intake during the previous day was recorded early in the morning of the following day. Symptoms of zinc deficiency, such as anorexia, diarrhea, dermatitis, and growth retardation, were observed. Two weeks later, the rats were sacrificed and serum zinc concentration was measured. Jejunal mucosa was taken for biopsy and was stained with hematoxylin and eosin (HE). The height ratio of the jejunal mucosal villi and crypts was measured. In addition, the activity of lactase in the jejunal mucosal brush border, gamma-glutamyl peptidase (GGT), and aminopeptidase N (APN) were measured. RESULT: The average weight of the rats in the ZA, ZD, and PF groups at the beginning of the experiment was (67.4 +/- 5.3) g, (64.7 +/- 4.8) g, and (66.5 +/- 4.1) g, respectively, and the average daily food intake was (11.2 +/- 1.0) g, (11.6 +/- 1.6) g, and (11.2 +/- 1.4) g, respectively. The intergroup differences were not significant. On the 7(th) day of experiment, no significant differences in average food intake were observed between the ZD group and the ZA and PF groups, but the average body weight in the ZD group was significantly lower than that in the ZA and PF groups (P < 0.01). At the end of the experiment (2 weeks), the average weight in the ZD group (112.0 +/- 11.5) g was significantly lower than that in the ZA (164.0 +/- 15.9) g and PF groups (137.5 +/- 16.2) g. The average food intake in the ZD group (13.4 +/- 5.1) g was significantly lower than that in the ZA group (18.2 +/- 2.4) g (P < 0.01). Serum zinc level in the ZD group (733 +/- 231) ug/L was significantly lower than that in the ZA (1553 +/- 159) ug/L and PF groups (1457 +/- 216) ug/L (P < 0.01). The height ratio of jejunal mucosa villus and crypt in the ZA, ZD, and PF groups was 2.98 +/- 0.5, 2.77 +/- 0.5, and 2.81 +/- 0.7, respectively, and lactase activity was (26.1 +/- 15.0) U/mg, (27.4 +/- 12.8) U/mg, and (40.8 +/- 18.5) U/mg, respectively, without significant intergroup differences. The GGT activity in the jejunal mucosa in the ZD group (12.7 +/- 6.5) U/g was significantly lower than that in the ZA (19.1 +/- 10.4) U/g and PF groups (18.5 +/- 7.7) U/g, but the difference was not significant. The activity of APN in the jejunal mucosa in the ZD group (25.5 +/- 7.5) U/g was significantly lower than that in the ZA (48.7 +/- 16.8) U/g and PF groups (43.9 +/- 14.5) U/g (P < 0.01). CONCLUSION: Zinc deficiency can cause loss of appetite, weight loss, and decreased activity of peptidase in the jejunal mucosal brush border. Zinc deficiency has little effect on the height ratio of the villus and crypt and lactase activity, thereby indicating that zinc deficiency may first affect protein digestion and absorption. PMID- 21624199 TI - [Motilin receptor expression in the interstitial cells of Cajal]. AB - OBJECTIVE: To look for the evidences of motilin receptor expression on interstitial cells of Cajal (ICC) of the rabbit. METHOD: Smooth muscle segments with ICC were isolated from the small intestine of 10-day old rabbits. The tissue segments equilibrated in Ca(2+)-free Hanks' solution were dispersed with an enzyme solution containing collagenase type II and then Ficoll density centrifugation was used to dissociate ICC. The cells were suspended and cultured in the M199 medium. The c-kit antibody was applied to distinguish the cultured ICC. The motilin receptor was identified by immunocytochemical assay with GPR38 antibody, c-kit antibody and hoechst 33342 combined to label ICC. Cells cultured for a few days were sorted for ICC with c-kit stained green fluorescent through flow cytometry. The total RNA and proteins extracted from the sorted ICC were respectively used to verify motilin receptor on the ICC by reverse-transcriptase polymerase chain reaction (RT-PCR) and Western blotting. RESULT: We had successfully dissociated and cultured ICC of rabbit small intestine in vitro. Fluorescent staining with c-kit antibody confirmed that the culture ICC was successful. Triple-labeled immunofluorescent staining had detected the motilin receptor on membrane of ICC. Flow cytometry analysis showed that the ratio of c kit positive cell in the cultured cells was 64.3%. The number of sorted ICC was 6.7 * 10(5) and 5.6 * 10(6). The results of RT-PCR and Western blot confirmed that the ICC had motilin receptor expression. CONCLUSION: Our study demonstrated presence of motilin receptor on ICC of the rabbit. The present results may suggest that ICC play an important role in gastrointestinal movement induced by motilin. PMID- 21624200 TI - [Expression of zinc finger protein A20 in pediatric inflammatory bowel disease]. AB - OBJECTIVE: It is demonstrated that excessive activation of NF-kappaB is central to the pathogenesis of inflammatory bowel disease (IBD). Zinc finger protein A20 (A20) is a key player in the negative feedback regulation of NF-kappaB signaling in response to multiple stimuli and has been described as central gatekeeper in inflammation and immunity. Mice genetically deficient in A20 develop severe intestinal inflammation and have increased susceptibility to dextran sodium sulfate (DSS)-induced colitis. Few studies have been done to explore the role of A20 in the pathogenesis of IBD. To clarify the relationship between intestinal inflammation and the expression level of A20 in IBD patients, the expression level of A20 and a series of inflammatory cytokines, such as NF-kappaB, IL-6, and IL-8, in children with IBD and controls were examined. METHOD: Terminal ileal mucosal samples were obtained via endoscopy. Fifty-seven mucosal samples were divided into 4 groups: normal control group (n = 16), IBD remission group (n = 12), IBD active group (n = 13) and non-IBD enteritis group (n = 16). According to disease activity index scores, the IBD patients were divided into IBD remission group and IBD active group. Normal control group was consisted of patients with functional bowel disorders or intestinal polyps. Non-IBD enteritis was defined as changes in which endoscopy and histological examination showed inflammatory changes but could not be diagnosed as IBD. Real-time PCR was adopted for detecting the mRNA levels of A20, IL-6 and IL-8. Meanwhile immunohistochemistry was performed to measure the expression of A20 and NF-kappaB. RESULT: (1) The expression of A20 and NF-kappaB were very low in normal control group, but significantly up-regulated in IBD active group and non-IBD enteritis group (P < 0.01 for both); (2) Compared with normal control group, expression of NF-kappaB [(9.35 +/- 4.84)% vs. (0.57 +/- 0.44)%, P < 0.01], IL-6 (t' = 1.34, P > 0.05), IL 8 (t = 1.38, P > 0.05) increased in IBD remission group, while the expression of A20 in both mRNA (t = 1.03, P > 0.05) and protein levels [(0.36 +/- 0.18)% vs. (0.87 +/- 0.29)%, P < 0.01] decreased; (3) Compared with non-IBD enteritis group, although the expression of NF-kappaB [(24.17 +/- 11.27)% vs. (55.29 +/- 21.84)%, P < 0.01], IL-6 (t = 2.22, P < 0.05), IL-8 (t = 2.97, P < 0.01) were highly increased in IBD active group, the expression of A20 in both mRNA(t = 2.26, P < 0.05) and protein levels [(29.23 +/- 11.70)% vs. (16.81 +/- 5.90)%, P < 0.01]significantly decreased; (4) The expression of IL-6, IL-8 were similar in IBD remission group and non-IBD enteritis group (both P > 0.05), but the expression of A20 was much lower in both mRNA (t = 4.42, P < 0.01) and protein levels [(29.23 +/- 11.70)% vs. (0.47 +/- 0.25)%, P < 0.01] in IBD remission group. CONCLUSION: The results demonstrate that there is an excessive inflammatory response but insufficient up-regulation of A20 expression in IBD patients. Low levels expression of A20 may play an important role in the pathogenesis of IBD. PMID- 21624202 TI - [Present status and prospect of diagnosis and treatment of inflammatory bowel diseases in children]. PMID- 21624201 TI - [Etiology and pathogenesis of inflammatory bowel diseases in children]. PMID- 21624203 TI - [Advances in molecular genetics of intestinal polyps in children]. PMID- 21624204 TI - [Influence of journals indexed by Science Citation Index (SCI) on Chinese medical journals based on the data of published articles by Chinese authors from 2000 - 2009]. AB - OBJECTIVE: This study was designed to investigate the influence of journals indexed by Science Citation Index (SCI) on Chinese medical journals. METHOD: Articles on medicine written by Chinese and the journals that published these articles from 2000 to 2009 were searched using Science Citation Index Expanded (SCI-E) database, and the status and variation tendency of the impact factors (IF) of these journals were analyzed. Data of articles on medicine included Chinese Scientific and Technical Paper and Citations Data (CSTPCD) from 2000 to 2008 were searched (the data of 2009 have not been released). The included articles and the time-dependent changing profile were studied. These outcomes were evaluated as the fixed base relative or link relative when compared with the data of 2000 or those of last year, respectively. Geometric mean was used when mean increase was calculated and IF distribution was described with median. RESULT: Totally 3774 articles from China were published by journals indexed by SCI-E in 2000, and the number of articles published by Chinese authors increased every year. In 2008, 16 714 articles were indexed by SCI-E, 442.87% higher than those of 2000. The increment was 161.54% higher than that of articles published in the journals indexed by CSTPCD (281.33%) during the same period. From 2000 to 2009, the geometric mean of increase in the number of published articles from China in journals indexed by SCI-E was 20.87% but it was 18.21% in CSTPCD. From 2000 to 2009, the median of IF of SCI-E indexed journals that published Chinese medical articles was 1.866, 2.073, 2.390, 2.702, 2.409, 2.496, 2.380, 2.218, 2.280 and 2.331, respectively, and they did not increase or even decreased. CONCLUSION: The number of the articles indexed by SCI-E increased year by year, much faster than that of CSTPCD. However, it does not necessarily mean the increase in impact. PMID- 21624206 TI - [Evaluation of the clinical and imaging examination in high-risk children with vesicoureteral reflux]. AB - OBJECTIVE: To analyze the clinical data and result of voiding cystourethrography (VCUG) in high-risk children with vesicoureteral reflux (VUR) for better awareness of VUR, and to assess the usefulness of non-radioactive voiding ultrasonography (VUS) in the diagnosis of VUR. METHOD: Ninety-three high-risk children with VUR who were hospitalized from July 2007 to April 2010 were studied. The study included 58 cases of urinary tract infection (UTI) and 35 cases of fetal or postnatal hydronephrosis detected on a B ultrasound scan. The results of urinalysis, urine culture, renal function, B ultrasound and VCUG were evaluated. Part of patients underwent VUS followed by VCUG immediately. RESULT: (1) Sixty-two boys and 31 girls (aged 1 month to 11.5 years, mean age 2 years) were included. VUR was detected in 26 patients (28%) by VCUG. In terms of kidney ureter units, VUR was detected in 36 of 186 kidney-ureter units, including 6 grade I, 3 grade II, 6 grade III, 15 grade IV and 6 grade V. (2) VUR was detected in 20 of 58 UTI patients (34.5%) by VCUG. The proportion of VUR in recurrent UTI group was 61.1%, much higher than that in first UTI group (22.5%). Thirteen of 20 VUR (65%) occurred in UTI patients under 1 year of age (M/F 10/3), with more bilateral VUR and severe grades of VUR than the older group. VUR was detected in 6 of 35 fetal or postnatal hydronephrosis patients (17.1%) by VCUG. (3) Twenty two patients underwent both VUS and VCUG. VUR was detected in 4 patients and 6 kidney-ureter units by VCUG, while in 6 patients and 9 kidney-ureter units by VUS. Taking VCUG as the reference standard, VUS had a sensitivity of 100%, specificity of 92.1%, positive predictive value of 66.7%, and negative predictive value of 100%. There was a concordance rate of 93.2% between VUS and VCUG. CONCLUSION: It is important to early screen VUR in UTI, fetal or postnatal hydronephrosis patients. There are more VUR, especially more bilateral VUR and severe grades of VUR, occurred in UTI patients under 1 year of age compared to older children. The incidence of VUR in recurrent UTI group was much higher than that in first UTI group. VUS is an accurate, reliable and radiation-free technique for the detection of VUR. It could be used to screen high-risk children for VUR and do the evaluation in the follow-up of VUR. PMID- 21624207 TI - [Meta-analysis of calcineurin inhibitor in the treatment of lupus nephritis]. AB - OBJECTIVE: To systematically evaluate the clinical effects of cyclosporine A (CsA) and tacrolimus, which are calcineurin inhibitors, on lupus nephritis. METHOD: In this study, the clinical trials on treatment of lupus nephritis with cyclosporine A and tacrolimus published until May 2010 were searched at www.guideline.gov, www.nice.org.uk, mdm.ca/cpgsnew/cpgs/index.asp, www.show.scot.nhs.uk, www.nzgg.org.nz, www.eguid elines.co.uk, www.gin.net, Cochrane library, EMBASE, MEDLINE, Wanfang database, Chinese Journal full-text Database, Chongqing Weipu Database by using the methods of Cochrane systematic review. At the same time the information from related journals, professional data and network were hand-searched. The homogeneous evaluation was performed by meta analysis.Statistical analysis of clinical data was performed by using RevMan 4.2 software provided by the Cochrane Collaboration. RESULT: A total of 214 reports were found, while only 7 randomized controlled trials met the inclusion criteria, 4 of them were on the treatment with CsA (treatment group) and cyclosporine (CTX) group (control group), and 3 of them were the on treatment with FK506 (treatment group) and CTX group (control group). There were 148 reports in the treatment of CsA and CTX group, while 185 reports in the treatment of FK506 and CTX group. Both CsA and tacrolimus group could decrease daily urinary protein. Tacrolimus group was good at reducing daily urinary protein as compared with CTX group, and the difference was statistically significant (Z = 2.8, P = 0.005), but there was no significant difference between CsA and CTX groups (Z = 1.08, P = 0.28). Tacrolimus group was good at complete remission as compared with CTX group (Z = 3.64, P = 0.0003), partial remission was similar in both groups (Z = 0.53, P = 0.6), and tacrolimus group was good at total remission (Z = 2.2, P = 0.03). There was no significant difference between CsA and CTX group in side effect within a short period, while FK506 had less side-effect than CTX group. CONCLUSION: Compared with the treatment with CTX, tacrolimus was good at reducing daily urinary protein. CsA and CTX were similar in reducing daily urinary protein in the treatment of lupus nephritis. Tacrolimus resulted in better total remission than CTX and had less side effect. CsA and CTX groups were similar in side effect. On the whole, calcineurin inhibitor could significantly decrease daily urinary protein, and tacrolimus was better in treatment and had less side-effect than CTX. However, large scale, multicenter, well-designed clinical trials should be adopted to further confirm the conclusions. PMID- 21624208 TI - [Blood lead status and influencing factors among preschool children in urban areas of China]. AB - OBJECTIVE: To investigate the blood lead status and influencing factors among preschool children in the sampling city. METHOD: Stratified-clustered-random sampling was used. Standardized questionnaire and peripheral blood samples were obtained from 69 968 children aged 0-6 years in fixed kindergartens and communities of Yinchuan, Xi'an, Chengdu, Wuhan, Hefei, Beijing, Harbin, Zhengzhou, Huhhot, Shijiazhuang, Haikou, Dalian, Qingdao, Guangzhou, Nanning and Changsha from 2004 to 2008, respectively. Tungsten atomic absorbtion spectrophotometry was employed to determine the blood lead level of children. RESULT: The proportion of children with blood lead level >= 100 ug/L was 7.57% (among which the proportion of high blood lead level, mild lead poisoning, moderate lead poisoning, severe lead poisoning were 91.0%, 2.76%, 3.32%, 2.93%, respectively) and the blood lead level was lower than those of the past studies. The proportion of high blood lead level has steadily declined from 2004 to 2008 [the proportions were 10.03%, 7.85%, 7.40%, 6.91% and 4.78%, respectively (chi(2) = 297.36, P < 0.0001)]. The proportion of children with blood lead level >= 100 ug/L in Haikou, Zhengzhou, Guangzhou, Shijiazhuang, Changsha, Xi'an, Wuhan, Hefei, Chengdu, Yinchuan, Harbin, Beijing, Dalian, Huhhot, Nanning and Qingdao were 12.15%, 10.49%, 10.37%, 9.69%, 9.53%, 9.46%, 9.40%, 8.50%, 7.99%, 7.98%, 7.51%, 6.10%, 3.25%, 2.89%, 2.46% and 2.39%, respectively (chi(2) = 768.21, P < 0.0001). By multiple regression method, the risk factors which influenced blood lead status of children were education status of mother, older children, behavior and dietary habit of children, boy, stay for long time in traffic busy areas, the type of housing, taking traditional Chinese and herbal medicine. The protective factors against lead poisoning in children mainly included scattered living, the nutritional status of calcium, iron, zinc, frequent intake of milk, and older mother. CONCLUSION: The blood lead level of children has decreased, but is still higher than those in developed countries. Lead exposure remains a public health issue which affects children most. The blood lead level of children is affected by multiple factors. Government and the whole society should pay attention to interrupt the lead pollutant and to promote nutritional health education. With all these efforts, it is possible to stop the progress of lead exposure and reduce its hazardous effects on the growth and development of children. PMID- 21624209 TI - [Two novel EIF2AK3 mutations in a Chinese boy with Wolcott-Rallison syndrome]. AB - OBJECTIVE: Wolcott-Rallison syndrome (WRS) is a rare autosomal recessive disorder characterized by the association of permanent neonatal or early-infancy insulin dependent diabetes, multiple epiphyseal dysplasia and growth retardation, and other variable multisystem clinical manifestations. Here we describe a Chinese boy affected by WRS. Genetic testing of his EIF2AK3 gene was performed in order to elucidate molecular variations and subsequently to provide credible genetic counseling for prenatal diagnosis in his family. METHOD: Based on analysis of a nine-year-old boy's clinical symptoms associated with biochemical examination and imaging, the diagnosis of WRS was therefore made. Genomic DNAs were extracted from peripheral blood leukocytes from the boy and his parents with their informed consent for genetic studies. All EIF2AK3 exons and intron-exon boundaries were amplified by Touch-down polymerase chain reaction (Touch-down PCR) and sequenced. RESULT: Direct sequencing of PCR products revealed the presence of a heterozygous T insertion (c.1408_1409insT) in exon 8 of the EIF2AK3 gene leading to frameshifting and termination, and another heterozygous T to A exchange (c.1596T > A) in exon 9 of the EIF2AK3 gene resulting in nonsense C532X mutation. CONCLUSION: Combining mutation screening of EIF2AK3 gene with clinical manifestations and effective examination may provide a reliable diagnostic method for patients. In this research, two novel mutations identified in the Chinese boy locate in the catalytic domain of the EIF2AK3 gene, disrupting the ability of autophosphorylation, leading to the truncated proteins that are unable to phosphorylate the natural substrate, which are responsible for the phenotype of Wolcott-Rallison syndrome. PMID- 21624210 TI - [Mutation analysis and prenatal diagnosis of 2 cases with mucopolysaccharidosis type I]. AB - OBJECTIVE: Mucopolysaccharidosis type I (MPS I; MIM# 252800) is an autosomal recessive disease that results from the deficiency in the lysosomal enzyme alpha L-iduronidase(IDUA). IDUA is one of the enzymes involved in degradation of glycosaminoglycans heparan sulphate and dermatan sulphate. The deficiency of IDUA leads to widespread accumulation of partially degraded mucopolysaccharides inside lysosomes, resulting in progressive cellular and multiorgan dysfunction. Up to now there is no definitely effective treatment for this disorder, therefore it is important to provide an accurate genetic diagnosis and prenatal diagnosis for the MPSI families. This study was conducted to detect IDUA gene mutation in patients with MPSIand make a definite diagnosis of homozygote or heterozygote and make first trimester prenatal diagnosis. METHOD: The 2 male probands included in this study were diagnosed as MPSI patients in Peking Union Medical College Hospital, case 1 was 2 years old and case 2 was 5 years old. Genomic DNA was extracted from leucocytes in the 2 patients and 2 mothers' cultured amniocytes. IDUA gene DNA sequence was amplified by polymerase chain reaction (PCR) and the PCR products were sequenced directly. Novel mutations were analyzed in 100 normal chromosomes. RESULT: The genotype of case 1 was p.L238R/c.883InsC, while of case 2 was c.531InsT/p.L346R. The fetal case 1 did not inherit the same pathogenic mutations as proband 1, the activity of the IDUA in amniocytes was 9.0 nmol/(h.mg pr). The fetal case 2 inherited the same pathogenic mutations with the proband, the genotype of fetal 2 was c.531InsT/p.L346R, the activity of the IDUA in amniocytes was 0.5 nmol/(h.mg pr). CONCLUSION: Of the 4 mutations found in 2 MPS I patients, p. L238R, c.883InsC, c.531InsT were novel. The fetal case 1 was diagnosed as normal fetus while the fetus 2 was diagnosed as affected. The results of the two kinds of prenatal diagnostic methods were correspondent with each other. PMID- 21624211 TI - [Clinicopathological feature and treatment of idiopathic membranous nephropathy in children]. AB - OBJECTIVE: To investigate the clinicopathological feature and treatment of idiopathic membranous nephropathy (IMN) in children. METHOD: A retrospective analysis of 25 cases of biopsy-proven IMN seen between January 2004 and December 2009. RESULT: The incidence of IMN was 3.81% in all the children patients who underwent renal biopsy. Of 25 patients with IMN, nine were boys and sixteen were girls. The mean age at onset was (9.4 +/- 3.4) years with a range of 2 - 14 years. Renal biopsies were performed at a median 2.5 months (range 0.4 - 11 months) after onset. The clinical manifestations included nephrotic syndrome (NS) nephritic type in 21 cases (84%) and glomerulonephritis in 4 cases. All patients presented with hematuria, and 7 had macroscopic hematuria. Hypertension was noted in 4 patients. Two patients were complicated with thrombosis. One patient was in a chronic renal insufficiency(CRI)state. According to the MN staging criteria, 21 cases were in stage II IMN (84%). Six patients showed moderate or severe tubulointerstitial lesion. Focal segmental glomerulosclerosis (FSGS) was found in two patients. Of the 22 patients with NS and nephrotic proteinuria, 21 cases were treated with prednisone initially and in 20 of them the efficacy of corticosteroid therapy was evaluated:one of them was steroid sensitive (became steroid-resistant after relapse) and all the others were steroid-resistant (95%). The subsequent treatment: eight of them were treated with prednisone followed by a taper to alternate-day therapy. Five of them had complete remission and three partial remission. Twelve cases were treated with combined therapy of prednisone and immunosuppressive agents. Of these 12 cases together with one case who received initially combined treatment with prednisone and immunosuppressive agent and one case treated with prednisone initially for five weeks then with combined therapy contained another immunosuppressive agent, totally 14 cases, 5 had complete remission, 2 partial remission, 3 did not achieve remission, and 3 had unknown response. CONCLUSION: Of the patient cohort, the predominant presenting feature was nephrotic syndrome, and with different degree hematuria. Almost all of them were steroid resistant, but followed by a taper to alternate-day therapy, some could achieve remission. The effect of a combination of prednisone and immunosuppressive agent is needed to be further proven in children. PMID- 21624212 TI - [Clinical detection of 22q11 microdeletion in the patients with congenital heart disease by multiplex ligation dependent probe amplification]. AB - OBJECTIVE: To detect 22q11 microdeletion in the children and fetuses affected by congenital heart defects. METHOD: MLPA P250 kit was used to detect 22q11 microdeletion in 100 cases of sporadic congenital heart defects including 40 fetuses and 60 patients diagnosed by ultrasound. RESULT: Two cases from the fetuses and 1 case from the patients were found to have 22q11 microdeletion. CONCLUSION: Three cases had 22q11 microdeletion in the congenital heart defects. PMID- 21624214 TI - [To advocate early intensive enteral nutrition for severe acute pancreatitis patients]. PMID- 21624213 TI - [Enlightenment from the gut-liver axis]. PMID- 21624215 TI - [Factors for the recurrence after endoscopic resection of early gastric cancer]. AB - OBJECTIVE: To investigate the related factors of recurrence of early gastric cancer (EGC) after endoscopic resection. METHODS: Clinicopathologic data of 169 patients with EGC who underwent endoscopic resection and periodically followed up by the Chinese PLA General hospital were analyzed retrospectively. RESULTS: During a follow-up of 13 - 57 months (median time 24.5 months), 12 patients had gastric cancer again and the recurrence rate was 7.1% (12/169). The recurrence time varied from 3 to 36 (28 +/- 23) months and the median time was 18 months. The recurrence rates of 0.5 year, 1(st) year, 2(nd) year and 3(rd) year were 1.18% (2/169), 3.55% (6/169), 9.91% (11/111) and 12.24% (12/98), respectively. Eleven patients had gastric cancer again within 2 years after resection. Undifferentiated histology (including poorly differentiated carcinoma and signet ring cell carcinoma), submucosal infiltration and lymphovascular invasion of the primary lesion of EGC were related to the postsurgical recurrence (all P < 0.05). CONCLUSION: Most recurrence of EGC occurred within 2 years after endoscopic resection and is related with undifferentiated histology, submucosal infiltration and lymphovascular invasion. It is important for these patients to receive endoscopy follow up. PMID- 21624216 TI - [The effects of enteral nutrition versus total parenteral nutrition on gut barrier function in severe acute pancreatitis]. AB - OBJECTIVE: To evaluate the effect of enteral nutrition (EN) versus total parenteral nutrition (TPN) on gut barrier function in patients with severe acute pancreatitis (SAP). METHODS: Sixty-three patients with SAP enrolled from 4 hospitals were randomly assigned into EN group (29 cases) and TPN group (34 cases). EN group patients were fed via a spiral nasojejunal feeding tube placed routinely by endoscopy or fluoroscopy, and TPN group patients were nourished intravenously with TPN during the same period. The changes of serum endotoxin, diamine oxidase, and urinary excretion of lactulose and mannitol ratio (L/M) were observed. RESULTS: Plasma concentration of endotoxin were markedly decreased in EN group as compared with that in TPN group at the 7(th), 14(th), 21(th) day of entry trial [(39.30 +/- 15.82) EU/L vs (73.05 +/- 21.16) EU/L, (22.64 +/- 14.31) EU/L vs (49.34 +/- 24.54) EU/L, (14.81 +/- 10.93) EU/L vs (30.08 +/- 14.10) EU/L, P < 0.05]. Plasma concentration of diamine oxidase were markedly decreased in EN group as compared with that in TPN group at the 7(th), 14(th) day of entry trial [(9.97 +/- 3.84) U/L vs (19.89 +/- 9.89) U/L, (5.42 +/- 1.84) U/L vs (8.79 +/- 4.08) U/L, both P < 0.05]. The urinary L/M decreased significantly in EN group than those in TPN group at the 7(th), 14(th), 21(th) day of entry trial (0.28 +/- 0.25 vs 0.65 +/- 0.45, 0.21 +/- 0.18 vs 0.54 +/- 0.41, 0.08 +/- 0.04 vs 0.29 +/- 0.06, all P < 0.05). CONCLUSION: EN has better effect on improving intestinal barrier function than TPN in treatment of patients with SAP. PMID- 21624217 TI - [The associations of methylenetetrahydrofolate reductase gene C677T and A1298C polymorphisms and ulcerative colitis]. AB - OBJECTIVE: To investigate the association between the genetic polymorphisms of methylenetetrahydrofolate reductase (MTHFR) and ulcerative colitis (UC) of Han ethnic population in Zhejiang, China. METHODS: Two hundred and seventy-four consecutive patients with UC and 726 healthy controls (HC) were studied. The genetic polymorphisms of MTHFR (C677T and A1298C) were genotyped using PCR-RELP methods. RESULTS: The frequencies of variant allele and genotype in MTHFR A1298C gene were higher in UC patients than in the HC (35.77% vs 29.96%, P = 0.013; 52.19% vs 44.90%, P = 0.039; respectively). However, there were no significant discrepancies of the allele and genotype frequencies in the MTHFR C677T gene between the UC patients and the HC (P > 0.05). In addition, the MTHFR 677TT homozygote, T allele and 677CT/1298AC compound genotype were more prevalent in patients with extensive colitis than in those with distal colitis (37.66% vs 14.72%, P = 0.0002; 49.35% vs 32.99%, P = 0.0004; 29.87% vs 15.23%, P = 0.006; respectively). Furthermore, the variant allele in the MTHFR A1298C gene (C) in severe UC patients was significantly lower than in mild and moderate UC patients (18.97% vs 33.88%, P = 0.022). CONCLUSION: The genetic polymorphisms of MTHFR C677T and A1298C are obviously associated with Han ethnic population with UC in Zhejiang province. PMID- 21624218 TI - [An analysis of intervention outcome in non-ST segment elevation acute coronary syndrome in elderly patients]. AB - OBJECTIVE: To investigate the effect and safety of early intervention and delayed intervention therapy on elderly patients and younger patients with non-ST segment elevation acute coronary syndrome. METHODS: The patients with non-ST segment elevation acute coronary syndrome were randomly divided into early intervention group (coronary angiography taken within 24 hours after grouping) and delayed intervention group (coronary angiography taken after 36 hours after grouping). The primary endpoint was a composite endpoint of death, myocardial infarction and stroke during 180 days follow-up. RESULTS: A total of 815 patients were enrolled, including 198 elderly patients aged 75 years and above, and 617 younger patients aged below 75 years. The elderly patients had a greater incidence of the primary endpoint than that of younger patients (P = 0.00). The primary endpoint of early intervention group were obviously lower than that of delayed intervention group of younger patients (P = 0.01). There was no significant difference in primary endpoint incidence of early intervention group and delayed intervention group of the elderly patients (P = 0.39). CONCLUSIONS: The elderly patients with non-ST segment elevation acute coronary syndrome who underwent intervention had greater incidence of death and myocardial infarction. Early intervention reduced the rate of myocardial infarction for the younger patients. There was no significant difference in primary endpoint incidence between early intervention and delayed intervention among elderly patients. PMID- 21624219 TI - [Monitoring of early Epstein-Barr virus reactivation and preemptive therapy after allogeneic hematopoietic stem cell transplantation]. AB - OBJECTIVE: To investigate early Epstein-Barr virus (EBV) reactivation and the outcome of preemptive therapy after allogeneic hematopoietic stem cell transplantation (allo-HSCT). METHODS: From January 2007 to January 2009, totally 277 patients after allo-HSCT were studied (haploidentical 116, unrelated 75, matched sibling 86). Conditioning regimens were mainly busulfan (BU) + cyclophosphamide (CY)/fludarabine (Flu) or total body irradiation (TBI) + CY/Flu. Antihuman thymocyte globulin (ATG) was added in haploidentical and unrelated transplants. Plasma EBV DNA was monitored once to twice weekly in the first 3 months after allo-HSCT with real time quantitative polymerase chain reaction (RQ PCR). EBV viremia was diagnosed when EBV DNA was more than 5 * 10(2) copies/ml but without symptoms. Acyclovir (10 mg/kg, intravenous drip, 8 h) was used for preemptive therapy and immuno-suppressants were decreased if possible. RESULTS: Totally 33 patients (11.9%) developed EBV viremia with a median time at day 44 (day 19 to day 84). The incidences of EBV viremia in the transplants from matched sibling, haploidentical, unrelated donors were 0, 15.5%, 20.0%, respectively. There was no significant difference between haploidentical and unrelated transplants (P = 0.09), but much less EBV viremia was seen in matched sibling transplant (P = 0.001). Twenty of 33 patients (60.6%) had complete response to preemptive therapy. The median time to reach EBV DNA negative in plasma was 11 (4 - 56) d. The median duration of preemptive therapy was 21 (14 - 60) d. Both univariate and multivariate analysis indicated that haploidentical and unrelated transplants, acute graft versus host disease (GVHD) were the risk factors for EBV viremia. Two-year overall survival in the patients with EBV viremia was significantly lower than that without EBV viremia (54.2% vs 72.1%, P = 0.006). CONCLUSIONS: Our large clinical study has demonstrated that preemptive therapy with acyclovir that is guided by EBV viremia is effective in majority of the patients with high-risk for EBV reactivation after allo-HSCT, which may further decrease the risk for developing life-threatening EBV disease or post transplantation lymphoproliferative disorder. Haploidentical and unrelated transplants, acute GVHD are the risk factors for EBV viremia which has negative impact on survival. PMID- 21624220 TI - [The relationship between obesity and microalbuminuria among general population in Beijing area]. AB - OBJECTIVE: To investigate the correlation between microalbuminuria (MAU) and obesity and its indexes, including BMI, waist circumference (WC), and waist-to hip ratio (WHR), among partial community population in Beijing. METHODS: A total of 2080 subjects who took physical examination in Beijing, including 810 men and 1270 women with a mean age of (50.9 +/- 13.1) years, were enrolled. The informed consent has been achieved from each patients. BMI and WHR were calculated based on collected data of height, weight, WC, and hipline. Urine albumin-creatinine ratio (ACR) within the range of 30 - 300 mg/g was classified as MAU. The subjects were divided into normal albuminuria (NAU) group and MAU group. The correlations between MAU and different obesity indexes including BMI, WC and WHR, were analyzed. RESULTS: Among the 2080 subjects, there was a positive correlation between BMI (r = 0.1276, P < 0.01) and ACR, and WC (r = 0.0840, P < 0.01) and ACR. WHR and ACR was irrelevant (P > 0.05). In univariate analysis, there was significant difference in BMI >= 28 kg/m(2) (OR = 2.02) and WC >= 85 cm (male) or >= 80 cm (female) (OR = 1.69) between NAU group and MAU group (P < 0.05). There was no significant difference in BMI 24- < 28 kg/m(2), and WHR >= 0.90 (male) or >= 0.85 (female) between NAU group and MAU group (P >= 0.05). Multivariate logistic regression analysis revealed that BMI (OR = 1.06) was an isolated independent risk factor of MAU from age (OR = 1.01), female (OR = 1.42), systolic blood pressure (OR = 1.01), TC (OR = 1.93) and HDL-C (OR = 0.54). CONCLUSIONS: Obesity is an independent risk factor of MAU among partial community population in Beijing. The correlation between different obesity indexes and MAU also differs. PMID- 21624221 TI - [Research on the effect of statins on insulin secretion from pancreatic islet in rats and its mechanisms]. AB - OBJECTIVE: To evaluate the inhibitory effect of statins on glucose-stimulated insulin secretion (GSIS) of pancreatic islet in rat and to explore its mechanisms. METHODS: According to the average volume, freshly isolated or 24-hour cultured pancreatic islets were randomly divided into control group (incubated with Kreb-Ringer bicarbonate buffer), the atorvastatin group (incubated with 100 umol/L atorvastatin), the fluvastatin group (incubated with 100 umol/L fluvastatin) and the pravastatin group (incubated with 100 umol/L pravastatin). Stimulated by 2.8, 5.5, 11.1, 16.7 mmol/L and 25.0 mmol/L glucose respectively, the effect of 100 umol/L statins on ATP content and GSIS was compared in the four groups. GSIS was performed by the 37 degrees C bath incubation method and ATP content was measured by chemiluminescence method. RESULTS: Incubated with 100 umol/L atorvastatin for 30 minutes, in the presence of 16.7 mmol/L glucose, the ATP content [(9.54 +/- 1.64) pmol/islet vs (12.33 +/- 1.89) pmol/islet] and GSIS (1.60 +/- 0.21 vs 2.39 +/- 0.30) were significantly reduced in comparison with the control group (P < 0.05). Cultured with 100 umol/L fluvastatin for 24 hours, the ATP content [(10.24 +/- 2.01) pmol/islet vs (12.31 +/- 2.16) pmol/islet] and GSIS (3.12 +/- 0.32 vs 4.17 +/- 0.37) were all significantly decreased at the higher glucose concentration of 16.7 mmol/L (P < 0.05). CONCLUSION: Atorvastatin and fluvastatin may inhibit GSIS by decreasing ATP content in pancreatic islet and the inhibitory effect is related to the strength of its lipophilicity. PMID- 21624222 TI - [The clinical analysis of 35 patients with cutaneous sarcoidosis]. AB - OBJECTIVE: To investigate clinical features of cutaneous sarcoidosis. METHODS: A retrospective analysis was carried out based on the clinic data of 35 patients with cutaneous sarcoidosis who were hospitalized in Peking Union Medical College Hospital during 1980 - 2009. They were divided into two groups, the group without systemic involvement (skin group) and the group with systemic involvement (systemic group). RESULTS: (1) The ratio of men and women with cutaneous sarcoidosis was 1:3.38, and the average incident age was (47.5 +/- 10.0) years old. The average incident age of skin group and systemic group were (41.8 +/- 12.5) years old and (50.5 +/- 7.1) years old, respectively. (2) The most common skin manifestation was subcutaneous nodule, followed by maculopapular and erythema nodosa. The most common involved sites were limbs. (3) The common involved systems extra-skin included the lung, joints and lymph nodes. The involvement rate of lung in cutaneous sarcoidosis of our present data was lower than those of foreign reports. However, the involvement rates of joints, lymph nodes, kidney, muscles and nervous system showed higher in our data. (4) The incidences of fatigue and weight loss in systemic group were higher than those in skin group (P < 0.05). The indexes of erythrocyte sedimentation rate, C-reactive protein and rheumatoid factor in systemic group were higher than those in skin group (P < 0.05). More patients in systemic group were treated with corticosteroid than that in skin group (95.7% vs 66.7%, P < 0.05). CONCLUSIONS: Subcutaneous nodules are the most common and the involvement rate of lung is lower in cutaneous sarcoidosis of our present data. Compared to the patients without systemic involvement, the average incident age of systemic ones is older, the indexes of inflammation markers and the usage of corticosteroid are higher. PMID- 21624223 TI - [The efficacy and safety of 2 cycles' high-dose dexamethasone treatment adult primary immune thrombocytopenia]. AB - OBJECTIVE: To investigate the efficacy and safety of a schedule of 2 cycles' high dose dexamethasone (HD-DXM) as an initial therapy in adults immune thrombocytopenia (ITP), and compare with conventional dose prednisone therapy. METHOD: A total of 59 newly diagnosed ITP patients were divided into 2 groups randomly. In 30 patients (Dexamethasone group), oral HD-DXM was administered at 40 mg/d for 4 consecutive days, repeated one week later, and then failed to maintain. In the remaining 29 patients (Prednisone group), prednisone was administered orally at 1.0 - 1.5 mg*kg(-1)*d(-1) for 4 weeks, and then gradually tapered. RESULTS: For short-term efficacy, after 1 and 2 weeks of treatment, the response rate in Dexamethasone group was significantly higher than that in Prednisone group (50.0% vs 24.1%, P < 0.01; 73.3% vs 55.2%, P < 0.05), while 3 weeks later, there was no remarkable difference between the two groups (83.3% vs 68.9%, P > 0.05), though the response rate in Dexamethasone group remained higher. For long-term effect, at the end of the 2nd and 3rd months of follow-up, the relapse rate in Dexamethasone group was significantly lower than that in Prednisone group (24.0% vs 40.0%, P < 0.05; 32.0% vs 65.0%, P < 0.01), while at the end of the 1(st) month of follow-up, there was no significant difference (16.0% vs 20.0%, P > 0.05). In addition, it's well tolerated and no complications such as severe infection or Cushing syndrome were complained in Dexamethasone group. CONCLUSION: HD-DXM possesses an advantage over conventional dose prednisone therapy in efficacy and safety. PMID- 21624224 TI - [The clinical significance of serum free light chain in primary systemic amyloidosis]. AB - OBJECTIVE: To evaluate the diagnostic and therapeutic significance of serum free light chain (sFLC) in primary systemic(AL) amyloidosis. METHODS: Twenty-five patients with AL amyloidosis, including 18 men and 7 women with a mean age of 54 (47 - 77) years old, were enrolled from October, 2005 to May, 2010. sFLC was measured by immunoturbidimetric assay. The type of monoclonal light chain was judged upon sFLC kappa/lambda and its sensibility was compared with serum immunofixation and immunohistochemical analysis. Four patients were treated with M(T)D (melphalan/thalidomide,and dexamethasone), one with VD (velcade and dexamethasone) and four with high-dose melphalan followed by autologous stem cell support. The changes of sFLC were serially determined before and after treatment. RESULTS: Among the 25 patients with AL amyloidosis, two were kappa light chains of precursor protein and 23 were lambda light chains. Mean plasma cell in bone marrow was 3.5% (0 - 15%). Nineteen (76%) patients had abnormal elevated sFLC and abnormal kappa/lambda ratios, and 17(68%) patients with immunofixation positive. The sFLC test had similar sensitivity as serum immunofixation (P = 0.727). Twenty one (84%) patients were shown to have either kappa or lambda immunoreactive amyloid deposits on biopsied tissues. The sFLC test combined with serum immunofixation allowed the M protein to be detected in 22 (88%) patients. The positive rates of immunohistochemical analysis combined with sFLC test and/or serum immunofixation were 96%. Four patients with hematologic response showed obvious improvement in visceral organ involvement, but illness of 5 patients without hematologic response kept stable or progressed. CONCLUSIONS: sFLC test is a sensitive qualitative and quantitative method to detect M protein. Preliminary data show the patients with obvious sFLC level decrease and/or kappa/lambda recovery to normal may have a high percentage of improved organs function. sFLC is critical index in diagnosing AL amyloidosis, which might help efficacy assessment. PMID- 21624225 TI - [The clinical analysis of nonaneurysmal subarachnoid hemorrhage]. AB - OBJECTIVE: To enhance recognition of the clinical and radiological features of nonaneurysmal subarachnoid hemorrhage (SAH) and its prognosis, and to provide guidance for the diagnosis and treatment in clinical practice. METHODS: Patients with spontaneous SAH, whose initial 3-dimensional digital subtraction angiography (DSA) were negative, received a second DSA after 2 to 3 weeks. Nonaneurysmal was diagnosed as SAH when both DSA were negative. All subjects were divided into 2 subgroups, perimesencephalic nonaneurysmal subarachnoid hemorrhage (PNSAH) group and nonperimesencephalic nonaneurysmal subarachnoid hemorrhage (n-PNSAH) group. RESULTS: Among 49 patients with nonaneurysmal SAH, 24 patients were PNSAH and 25 patients were n-PNSAH. Two patients died and 47 patients recovered and discharged with an average follow-up of 26 months. CONCLUSION: The clinical course and prognosis of patients with PNSAH were good, better than that of patients with n PNSAH. PMID- 21624226 TI - [Functional MRI and cognition assessment in subcortical ischemic vascular disease]. AB - OBJECTIVE: To evaluate the imaging features of patients with vascular cognitive impairment (VCI) induced by Subcortical Ischemic Vascular Disease (SIVD), through magnetic resonance diffusion tensor imaging (DTI) and proton spectroscopy (MRS) technology. METHODS: A total of 52 patients with SIVD were enrolled. After analysis of scale score, 32 patients with cognitive impairment were assigned to VCI group and 20 patients with no cognitive impairment were assigned to control group. Both group received DTI and MRS examination. The mean values of apparent diffusion coefficient (ADC) and fractional anisotropy (FA) of the bilateral temporal, frontal, parietal and occipital white matter regions as well as in the bilateral centrum semiovale were calculated. The peak value of MRS of N-acetyl aspartate (NAA), choline (Cho), creatine (Cr) and phaseomannite (mI) were calculated. RESULTS: Compared with control group, FA decreased in the region of temporal, frontal, parietal as well as in the centrum semiovale, and ADC increased in VCI group (P < 0.05). In the frontal regions and centrum semiovale, the VCI patients had a significant FA decrease. The ADC value increased obviously in the temporal lobe. Spectrum analysis results showed, NAA/Cr was lower than control group in VCI group in the frontal lobe (1.43 +/- 0.08 vs 1.53 +/- 0.92), while mI/Cr was higher than control group in the temporal lobe (0.51 +/- 0.06 vs 0.46 +/- 0.07) (P < 0.05). CONCLUSION: FA in the temporal and centrum semiovale regions of VCI group and NAA in the temporal white matter regions decreased obviously. DTI and MRS could provide a reference value for early diagnosis and assessment of VCI. PMID- 21624227 TI - [A study of serum cartilage oligomeric matrix protein and matrix metalloproteinase-3 concentration in osteoarthritic rabbit models]. AB - OBJECTIVE: To study the levels of cartilage oligomeric matrix protein (COMP) and matrix metalloproteinase-3 (MMP-3) in the serum fluid of osteoarthritic rabbit models and their relationships with the severity of pathological changes, so as to investigate their correlation with osteoarthritis (OA). METHODS: The osteoarthritic animal models were get from immobilizing the right knees of 18 rabbits in full extension using plaster cast. Knee joint pathological changes of 2, 6 weeks were examined for pathological severity of OA; ELISA sandwich method was used to measure the levels of COMP and MMP-3 in serum before and after modeling (at 2, 6 weeks respectively); X ray of model keens was also obtained in different period. Correlation analysis was performed to demonstrate the relationship between the levels of COMP, MMP-3 in the serum and the pathological severity of OA. RESULTS: (1) Morphological observations:immobilizing the right knees of rabbits in full extension using plaster cast was a reliable method for osteoarthritic animal models and the typical histopathologic character was seen; the severity of osteoarthritis gradually increased with time extended. (2) The levels of COMP [(3.64 +/- 0.18) ug/L], MMP-3 [(1.99 +/- 0.81) ug/L] in the serum of 2 weeks osteoarthritic animal models were higher than those before immobilizing with plaster cast [COMP (3.35 +/- 0.20) ug/L, MMP-3 (1.61 +/- 0.71) ug/L]. The levels of COMP [(3.96 +/- 0.44) ug/L], MMP-3 [(3.44 +/- 0.91) ug/L] of 6 weeks were much higher, with a significant difference (P < 0.05). The levels of COMP, MMP-3 in serum had a linear correlation with the pathological severity of OA (r > 0.710, and P < 0.05). CONCLUSION: The levels of COMP and MMP-3 in serum can help to predict and evaluate the progression of OA. PMID- 21624228 TI - [The effects of Berberine on growth of Immorto-Min colonic epithelial cell line]. AB - OBJECTIVE: To investigate the effects of Berberine on growth of Immorto-Min colonic epithelial cell line (IMCE) and explore its possible mechanisms. METHODS: IMCE cells were treated with Berberine in the absence or presence of epidermal growth factor (EGF) and TNFalpha. Ki-67 staining and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay were used to identify the cell proliferation and apoptosis respectively. Furthermore, Western blot analysis was performed to detect the epidermal growth factor receptor (EGFR), protein kinase B (Akt) and their phosphorylation. RESULTS: (1) Proliferating activity of IMCE cells was increased after adding EGF and the proportion of cell proliferation was (10.64 +/- 1.41)%. The proportion was significantly lowed in EGF plus Berberine group [(1.81 +/- 0.85)%] compared to the EGF group (P < 0.01), while the lowest was the Berberine group [(0.49 +/- 0.42)%]. (2) The proportions of cell apoptosis were (8.47 +/- 2.52)% and (9.39 +/ 2.13)% in the Berberine group and TNFalpha group respectively which were significantly higher compared to the normal control [(0.27 +/- 0.30)%], both P < 0.01. (3) The phosphorylation of EGFR was significantly increased after adding EGF and p-EGFR was decreased in EGF plus Berberine group at a concentration dependent manner. (4) Moreover, the phosphorylation of Akt was enhanced after addition of TNFalpha, while the phosphorylation in the TNFalpha and Berberine group was inhibited compared to the TNFalpha group. CONCLUSIONS: Berberine may suppress the proliferation and promote the apoptosis of IMCE cells. The mechanisms may relate to the inhibition of the phosphorylation of EGFR and Akt. PMID- 21624229 TI - [To raise awareness on the importance of multiple etiology joint detection in acute respiratory infections]. PMID- 21624230 TI - [Characteristics and the prevalence of respiratory viruses and the correlation with climatic factors of hospitalized children in Suzhou children's hospital]. AB - OBJECTIVE: To study the epidemiological characteristics of respiratory virus infection and its relations to climatic factors in Suzhou. METHODS: From 2006 to 2009, viral etiology surveillance was conducted among 6655 children hospitalized with acute respiratory tract infections (ARIs). Direct immunofluorescence method was used to test respiratory secretion samples for respiratory syncytial virus (RSV), influenza viruses A and B (Inf-A, Inf-B), parainfluenza virus types I, II, and III (Pinf-I, Pinf-II, Pinf-III) and adenovirus. Samples were tested for human metapneumovirus (hMPV) with reverse transcription polymerase chain reaction (RT PCR). Samples from Jan 2006 to Dec 2009 were also tested for human bocavirus (HBoV). Climatic factors, including mean temperature, relative humidity, rainfall amount, sum of sunshine and mean wind velocity were collected monthly. The relationship between activity of each virus and climatic factors were analyzed by linear regression and stepwise regression analysis. RESULTS: From 2006 to 2009, in the total virus detection rate was 32.2% (2142/6655) in Suzhou. RSV was the most common virus and the average detection rate was 15.7% (1048/6655), followed by hMPV 8.9% (596/6655), HBoV 7.8% (148/1883), Pinf-III 2.7% (183/6655), Inf-A 2.4% (161/6655), ADV 1.3% (89/6655), Pinf-I 0.4% (29/6655), Inf-B 0.37% (25/6655) and Pinf-II 0.16% (11/6655). The positive rates of RSV, hMPV and ADV were significantly different in four years (chi(2) = 17.71, 33.23, 8.42, all P values < 0.05). Different virus has different epidemiological characteristics and distinct seasonality. The detection rate of RSV, hMPV, Inf-A were higher in Winter as 37.2%, 13.2%, 4.4%, respectively. ADV and Pinf-III were higher in summer as 2.3% and 4.6% respectively. The peak of HBoV existed in Autumn as 3.3%. The total virus detection rate showed significant inverse correlation with month average temperature (r = -0.732, P < 0.001) and a weak inverse correlation with average wind velocity was also found (r = -0.36, 0.01 < P < 0.05). The highest month total virus detection rate was from 47.6% to 84.4% when average temperature was from 3.2 degrees C to 9.4 degrees C and mean wind velocity was from 1.2 - 1.9 m/s. The associations of average temperature, sum of sunshine and wind velocity with RSV activity were statistical significant (r = -0.88, P < 0.001; r = -0.43, P < 0.01; r = -0.47, P < 0.01). The highest rate was from 24.3% to 58.2%, when mean temperature was from 5.3 degrees C to 19.9 degrees C, mean wind velocity was from 1.3 - 2.4 m/s and sum of sunshine was 61.0 to 153.4 hours. hMPV detection rate was inversely correlated with mean temperature and rain account (r = -0.43, P < 0.01; r = -0.29, P < 0.05). The rate was highest from 11.7% to 31.6% when mean temperature was from 5.3 degrees C to 21.9 degrees C and rain account was from 27.5 millimeter to 150.9 millimeter. Only mean temperature was positively correlated with Pinf-III (r = 0.53, P < 0.001). The rate was from 2.8% to 7.2% when mean temperature was between 11.9 degrees C and 30.4 degrees C. ADV detection rate was positively correlated with mean temperature and sum of sunshine, but negatively correlated with wind velocity (r = 0.35, P < 0.05; r = 0.30, P < 0.05; r = -0.32, P < 0.05). The rate was from 2.2% to 6.6% when mean temperature was between 15.9 degrees C and 30.4 degrees C, and sum of sunshine between 93 hours to 240.7 hours and mean wind velocity was from 1.1 - 2.8 m/s. Average temperature and relative humidity showed interactions on the detection rate of ADV (r = 0.36, P = 0.0093; r = -0.34, P = 0.016), but temperature showed higher effect on ADV detection rate. ADV detection rate was high at higher temperature (15.9 - 30.4 degrees C) and low humidity (56% - 71%). CONCLUSION: RSV was one of the most common viruses among hospitalized children in Suzhou, and hMPV and HBoV also played an important role in respiratory tract infection of children. Different virus has different cycle and seasonality. Climatic factors, especially mean temperature, was the main factor affecting the virus prevalence. PMID- 21624231 TI - [Comparative studies on the composition and antibiotic-resistance of pathogenic bacteria between children with community-acquired and hospital-acquired pneumonia]. AB - OBJECTIVE: This research was to explore the difference between children with community-acquired pneumonia (CAP) and hospital-acquired pneumonia (HAP) in the composition and antibiotic-resistance of pathogenic bacteria. METHODS: 241 CAP and 116 HAP with positive sputum culture who were hospitalized from January to December in 2008 in Children's Hospital Affiliated to Suzhou University were selected in this study. The bacteria were identified by traditionally manual method and antibiotic sensitivity tests were performed by K-B method. The chi square or Fisher's exact test were used for statistical test. RESULTS: In 241 CAP, Streptococcus pneumoniae and haemophilus influenza accounted for (42.2%, 106/251) and (12.4%, 31/251) infection, respectively; however in 116 HAP, Enterobacteriaceae and Non-fermenters accounted for (88.2%, 127/144). In addition, methicillin-resistant Staphylococcus aureus weren't isolated, however, its detection rate was 66.7% in HAP. The drug resistance was 1.5 times higher in HAP than that in CAP for several types of antibiotics, such as ceftazidime (37.5% (6/16) vs 75.6% (31/41)), cefepime (37.5% (6/16) vs 78.0% (32/41)), aztreonam (50.0% (8/16) vs 90.2% (37/41)), cefoperazone/sulbactam (12.5% (2/16) vs 51.2% (21/41)) and piperacillin/tazobactam (12.5% (2/16) vs 56.0% (23/41)). Klebsiella pneumoniae isolated from HAP had higher drug resistance than that isolated from CAP against some antibiotics, for example, gentamicin (0 vs 63.6% (7/11)), SMZ + TMP (20.0% (1/5) vs 63.6% (7/11)) and cefoperazone/sulbactam (0 vs 54.5% (6/11)). We also found Enterobacter cloacae isolated from HAP showed high drug resistance than that isolated from CAP against imipenem (0 vs 46.7% (7/15)), aztreonam (9.1% (1/11) vs 60.0% (9/15)) and cefoperazone (18.2% (2/11) vs 80.0% (12/15)) and Pseudomonas aeruginosa from HAP had higher resistance than that from CAP against gentamicin (0 vs 50.0% (9/18)), amikacin (0 vs 38.9% (7/18)), ceftazidime (0 vs 55.6% (10/18)), cefepime (0 vs 50.0% (9/18)) and cefoperazone (33.3% (2/6) vs 94.4% (17/18)). The detection rates of ESBLs for Escherichia coli were 84.6% (11/13) and 93.3% (14/15) in CAP and HAP, respectively (chi(2) = 0.553, P > 0.05); while for Klebsiella pneumoniae, they were 81.3% (13/16) and 95.1% (39/41), respectively (chi(2) = 2.767, P > 0.05). CONCLUSION: CAP was mainly comprised of Streptococcus pneumoniae and haemophilus influenza; while HAP was mainly comprised of Enterobacteriaceae and Non-fermenters. The drug resistance of gram-negative bacilli was higher in HAP than that in CAP. PMID- 21624232 TI - [Analysis of antibiotic resistance and genotypes on extended spectrum beta lactamase and AmpC beta-lactamase producing strains isolated from Uygur and Han newborns]. AB - OBJECTIVE: This study aimed to investigate drug resistance and genotypes of the extended spectrum beta-lactamase (ESBLs) and AmpC beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae isolated from Uygur and Han newborns in Urumqi. METHODS: Disk diffusion test (Kirby-Bauer) was used for detecting drug resistance of 299 strains to twenty two kinds of antibiotics. Resistance genes of the ESBLs and AmpC beta-lactamase-producing strains were amplified by multiplex PCR and subtypes were confirmed by DNA sequence analysis. Total 148 strains were selected with random number table and sequenced, which included TEM-, SHV-, CTX-M 1-, or CTX-M-9-positive ESBLs-producing strains and DHA-, or CIT-positive AmpC beta-lactamase-producing strains. Antibiotic resistant rates were analyzed by Whonet 5.4 and statistic analysis was performed by chi-square (chi(2)) test with PEMS 3.1. RESULTS: The antibiotic resistant rates between Uygur and Han newborns significantly differ in ESBLs-producing Klebsiella pneumoniae to Sulfamethoxazole Trimethoprim (80.0% (40/50) and 56.0% (28/50), chi(2) = 6.6176, P = 0.0101), in ESBLs-producing Escherichia coli to Sulbactam and Cefoperazone (54.2% (32/59) and 94.0% (47/50), chi(2) = 21.4512, P = 0.0000), and in AmpC beta-lactamase producing Klebsiella pneumoniae to Sulbactam and Cefoperazone (100.0% (20/20) and 72.2% (26/36), chi(2) = 6.7633, P = 0.0093) and to Amikacin (65.0% (13/20) and 25.0% (9/36), chi(2) = 8.6246, P = 0.0033). Although SHV gene of ESBLs-producing Escherichia coli was detected from Uygur newborns at only 3.4% (2/59) and not detectable from Han newborns, TEM, CTX-M-1, and CTX-M-9 group genes were all detected over 38.0% (19/50). Among the detected strains, the subtypes of TEM and CTX-M-1 were mainly TEM-1 and CTX-M-15, respectively; whereas the subtypes of SHV and CTX-M-9 included SHV-1, 2, 11, 12, 27, 61, 99 and CTX-M-9, 14, 24, 27, 65, respectively. The strains of Escherichia coli and Klebsiella pneumoniae carrying two or more kinds of ESBLs genotypes were 56.7% (42/74) - 90.0% (63/70). Two species carrying the AmpC gene in two kinds of newborns were only grouped in the subtypes of DHA-1 and CMY-44, and other subtypes were not detected at all. Moreover, TEM-positive ESBLs-producing Escherichia coli were detected from Uygur newborns at the higher rate than that from Han newborns (71.2% (42/59) and 50.0% (25/50), chi(2) = 5.1291, P = 0.0235), while there was no difference in other genotypes detected between two kinds of newborns (chi(2) < 3.7780, P > 0.05). CONCLUSION: There were significant differences in antibiotic resistance and genotype distribution of Klebsiella pneumoniae and Escherichia coli between two nationality newborns, and these two bacteria detected in this study carried multi resistance genes and showed high resistant to beta-lactamase antibiotics. PMID- 21624233 TI - [Analyzing the mutations of rpoB gene in Mycobacterium tuberculosis clinical isolates by probe melting analysis assay]. AB - OBJECTIVE: To evaluate the clinical performance of a probe melting analysis (PMA) based real-time PCR detection kit in rapid detection of rifampin-resistant mutations in Mycobacterium tuberculosis (MTB). METHODS: The specificity of the assay was evaluated by detecting 37 non-tuberculous mycobacteria (NTM), and the detection limit of the method was evaluated by genomic DNA of a standard strain H37Rv. Finally, 962 clinical isolates were analyzed with the PMA assay by detecting mutations in rifampin resistance-determining region (RRDR) of rpoB gene, and results were verified with DNA sequencing. RESULTS: Among 37 NTM strains, three strains showed drug resistant mutation signals. The PMA method could detect down to 30 bacteria per reaction. Sample analysis showed that 186 of 962 isolates were mutants, 751 isolates were wild type and 25 isolates failed to give amplification signals. Among the mutant samples detected, 112 samples from November 2009 to April 2010 were further analyzed by sequencing, as well as 200 wild-type samples. The results showed a complete agreement with the PMA assay except for 5 samples failed in sequence analysis. CONCLUSION: The PMA assay is rapid, accurate and easy-to-use, and thus can be used for detection of rifampin resistant in clinical isolate samples. PMID- 21624234 TI - [Preliminary study on bacteroides as the potential fecal contamination indicator bacteria]. AB - OBJECTIVE: To explore the possibility of Bacteroides spp. as fecal contamination indicator bacteria with real-time quantitative PCR (RT-PCR) assay through analyzing the correlation between Bacteroides spp. and coliform group in external environment. METHODS: Quantity of coliform group and Bacteroides in water samples were detected by most-probable-number method (MPN) and RT-PCR, respectively, and their detection correlation was evaluated with linear correlation analysis. Both methods were also applied to detect the contaminated time limits and river water samples collected at four sampling sites in three different times. RESULTS: Seventy two hours were needed for the numeration of coliform group with MPN method, while RT-PCR could detect Bacteroides within 3 hours. The contaminated time limit of indoor and outdoor water samples of coliform group was more than 40 days and 9 days, and Bacteroides 13 days and 5 days, respectively. Also, the positive correlation between the quantity of Bacteroides and coliform group in outdoor water samples was obtained, the quantity of Bacteroides was from 8.3 * 10(6) copies/ml to less than 10(4) copies/ml during the first day to the fifth day, while coliform group was 4.3 * 10(6) MPN/100 ml to 2.4 * 10(3) MPN/100 ml. A 100% coincidence rate of the detection results with both methods was also observed. These results indicated that the detection results of both methods had perfect consistency. CONCLUSION: Bacteroides spp. can be potentially used as fecal contamination indicator bacteria with RT-PCR rapid detection. PMID- 21624235 TI - [A study on indicator system for early-warning on hemorrhagic fever with renal syndrome epidemic]. AB - OBJECTIVE: To establish an early-warning indicator system on outbreak of hemorrhagic fever with renal syndrome by Delphi method seeking expert advices. METHODS: Firstly, the literature review and the experts meeting method were used to formulate the initiator frame work and indicators. A two-round consultation was used to filter indicators, discuss the boundary of indicators and determine the weighting coefficient among 25 experts from 14 provinces, municipalities and autonomous regions. The relative weightiless of indicators was determined by the weight coefficients method. RESULTS: The experts' average length of service in prevention and control of hemorrhagic fever with renal syndrome was (23.80 +/- 11.70) years. The positivity coefficients of the two-round experts were 100% and 72%. Kendall's coefficients of the two-round consultation were 0.50 (chi(2)(R) = 148.95, P < 0.01) and 0.54 (chi(2)(R) = 212.63, P < 0.01) and opinions among experts became consistent and the consultation had achieved the need of forecast. Four first-class indicators (host animals, risk population, social environment and case-related indicators) and 14 second-class indicators were filtered to develop the indicators system. The weight coefficients of the first-class indicators were 0.28, 0.23, 0.23 and 0.26. CONCLUSION: The early-warning index system of hemorrhagic fever with renal syndrome has been established and it could provide a reference for the forest and warning of HFRS outbreak. PMID- 21624236 TI - [Relationship between genetic polymorphism in microRNAs precursor and genetic predisposition of hepatocellular carcinoma]. AB - OBJECTIVE: To investigate the relationship between genetic polymorphism in microRNAs (miRNAs) precursor and genetic predisposition of hepatocellular carcinoma (HCC) in Chinese population. METHODS: A case-control study including 963 HCC cases and 829 HBsAg positive controls and 852 HBsAg negative controls was conducted. hsa-mir-146a rs2910164 C->G and hsa-mir-196-a2 rs11614913 T->C were selected, where the genotypes were determined by the primer introduced restriction analysis-PCR (PIRA-PCR) assay. Odd ratios (ORs) and 95% confidence intervals (CIs) were evaluated by logistic regression analysis to investigate the relationship between onset risk of HCC and different genotypes. RESULTS: The genotype frequencies of CC, CG and GG at rs2910164 gene locus were separately 34.5% (319/925), 48.6% (450/925) and 16.9% (156/925) in cases; 36.4% (274/753), 45.0% (339/753) and 18.6% (140/753) in HBsAg positive controls; and 36.1% (303/840), 46.0% (386/840) and 18.0% (151/840) in HBsAg negative controls. The genotype frequencies of TT, CT and CC at rs11614913 were respectively 29.7% (277/934), 48.1% (449/934) and 22.3% (208/934) in cases; 30.3% (238/785), 51.0% (400/785) and 18.7% (147/785) in HBsAg positive controls; and 28.6% (239/837), 49.8% (417/837) and 21.6% (181/837) in HBsAg negative controls. No significant relationships were observed between these two single nucleotide polymorphisms (SNPs) and onset risk of HCC after adjusting the factors as age, gender, smoking and drinking status in comparison with HBsAg positive controls: hsa-mir-146a rs2910164 (CG + GG vs CC): adjusting OR = 1.10, 95%CI: 0.90 - 1.36; hsa-mir-196 a2 rs11614913 (CC + CT vs TT): adjusting OR = 1.01, 95%CI: 0.81 - 1.25; as well as in comparison with HBsAg negative controls: hsa-mir-146a rs2910164 (CG + GG vs CC): adjusting OR = 1.06, 95%CI: 0.87 - 1.29; hsa-mir-196-a2 rs11614913 (CC + CT vs TT): adjusting OR = 0.94, 95%CI: 0.76 - 1.16. As well, no significant relationships were observed between these two SNPs and onset risk of HCC in the subgroups stratified by age, gender, smoking and drinking status. CONCLUSION: hsa mir-146a rs2910164 C->G and hsa-mir-196-a2 rs11614913 T->C may not play an important role in the HCC predisposition among Chinese populations. PMID- 21624237 TI - [Analysis of risk factors for upper gastrointestinal cancer in China: a multicentric population-based case-control study]. AB - OBJECTIVE: To explore the major risk factors for upper gastrointestinal cancer in high occurrence areas of esophageal and gastric cancer in China. METHODS: Four high occurrence areas of esophageal cancer, namely Cixian and Shexian from Hebei province, Linxian from Henan province, Feicheng from Shandong province, and Zhuanghe from Liaoning province, which is a high occurrence area of gastric cancer, were selected for the study. The newly-diagnosed cases whose date of onset were after January 1st, 2009 were selected from the Cancer Registration Database in each district, and 751 cases diagnosed as cancers in lower segment of esophagus, cardiac and other subsite of stomach were randomly recruited. 2253 matched controls were selected to pair the cases at the ratio of 3:1. The relative information of the study objects were collected from the face-to-face interviews with trained staff by designed questionnaires, and the data was input by EpiData software. Statistic software SPSS 13.0 was applied to conduct both univariate and multivariate logistic regression analysis to evaluate odd ratios (OR) and 95% confident interval (CI). RESULTS: As univariate analysis shown, 66 objects in case group had irregular diet habit; while 90 in control group had (OR = 3.177; 95%CI: 2.127 - 4.745). A higher percentage in case group (83 objects) preferred fried food in comparison with only 214 in control group did (OR = 3.190; 95%CI: 2.061 - 4.927). 369 objects in case group, but only 119 in control group had history of gastrointestinal diseases (OR = 14.660; 95%CI: 11.342 - 18.948). 282 objects in case group had history of gastroesophageal reflux disease (GERD), which was much higher than the percentage in control group (432 objects), with OR = 3.137 (95%CI: 2.546 - 3.864). All the above factors could increase the risk for upper gastrointestinal cancer. 387 objects in case group and 1278 in control group reported they preferred fresh vegetables in daily diet, which was found to be a protective factor (OR = 0.609; 95%CI: 0.473 - 0.785). As multivariate analysis shown, history of gastrointestinal tract diseases (OR = 21.420; 95%CI: 15.484 - 29.632), irregular food diet (OR = 3.097; 95%CI: 1.740 - 5.514), pickled food (OR = 3.005; 95%CI: 1.873 - 4.819), and GERD (OR = 2.261; 95%CI: 1.673 - 3.057) were found to be risk factors for upper gastrointestinal cancer; while frequent fresh-vegetable diet was a protective factor (OR = 0.562; 95%CI: 0.396 - 0.800). CONCLUSION: Irregular lifestyle and unhealthy diet habit could be the major risk factors for upper gastrointestinal cancers among the residents from high occurrence areas of esophageal cancer and gastric cancer in China. PMID- 21624238 TI - [Incidence trends and pathological characteristics of lung cancer in urban Beijing during period of 1998 - 2007]. AB - OBJECTIVE: To describe the incidence trends and pathological characteristics of lung cancer in urban Beijing, China. METHODS: A total of 32 845 medical records of the residents diagnosed as lung cancer in urban Beijing from 1998 to 2007 were retrieved through the cancer registry system of Beijing Cancer Registry. Crude incidence rate, age-specific incidence rate, adjusted incidence rate by world standardized population, annual percentage change (APC) and histological categorized incidence rate by world standardized population were calculated in order to compare the differences of the incidence trends in different time periods, or among different gender and age groups. RESULTS: A total of 32 845 newly diagnosed lung cancer patients between 1998 and 2007 were included in our study. The crude incidence rate was 47.81/100 000 (32 845/68 704 429), increasing by 38.80% from 39.30/100 000 in 1998 to 54.55/100 000 in 2007 with APC at 3.35% in urban Beijing (Z = 9.984, P < 0.001). While it changed to 28.95/100 000 with an APC at 0.27% (Z = 0.846, P = 0.422) when adjusted by world standardized population. For male, the crude incidence rate was 58.28/100 000 (20 342/34 906 580, adjusted rate at 37.03/100 000, APC at 0.38%, Z = 1.008, P = 0.343); while for female, the crude incidence rate was 36.99/100 000 (12 503/33 797 849, adjusted rate at 21.48/100 000, APC at 0.14%, Z = 0.431, P = 0.678). 17 920 lung cancer patients being diagnosed according to histological evidence, accounted for 54.56%. The respective proportion of the patients with histological diagnosis was 43.14% (1095/2538) in 1998 and 65.55% (2641/4029) in 2007, with a 51.95% increase (chi(2) = 859.152, P < 0.001) in decade. In terms of subtypes of lung cancer, the proportion of squamous cell carcinoma decreased annually, from 30.41% (333/1095) in 1998 to 24.16% (638/2641) in 2007; while the proportion of adenocarcinoma increased from 42.83% (469/1095) to 46.80% (1236/2641). As a result, the squamous cell carcinoma to adenocarcinoma ratio declined from 0.71 (333/469) to 0.52 (638/1236) (chi(2) = 50.214, P < 0.001). For women, the ratio declined more significantly and the proportion of the squamous cell carcinoma and adenocarcinoma were 14.77% (925/6262) and 60.83% (3809/6262), respectively in the period between 1998 and 2007. CONCLUSION: No significant change was found in the incidence trend of lung cancer after the incidence rate adjusted by world standard population, but the proportion of the subtypes of lung cancer categorized by histological evaluation changed apparently. PMID- 21624239 TI - [Case-control study of the relationship between children's moderate-severe asthma and development indicators]. AB - OBJECTIVE: To investigate the relationship between moderate-severe asthma and development in children. METHODS: A total of 389 children in the state of moderate-severe persistent asthma were enrolled, which were divided into moderate (226 cases) and severe group (163 cases). According to age, each group was divided into three groups: 3-year-old (85 cases and 63 cases), 5-year-old (76 cases and 52 cases) and 7 to 10 year-old (65 cases and 48 cases). Meanwhile, 298 healthy children in the same age group were enrolled as control, of which 3-year old were 96 cases, 5-year-old were 92 cases and 7 to 10 year-old were 110 cases. Height, weight and lung function were measured respectively. RESULTS: 3-year-old of severe group, the value of height, the value of weight, the percentage of height, the percentage of weight, the SDS of height, the SDS of weight [(98.54 +/ 7.75) cm, (14.87 +/- 2.46) kg, 50.30% +/- 16.31%, 50.27% +/- 18.29%, 0.11 +/- 0.66, 0.06 +/- 0.49, respectively] were lower than the moderate group of the same age group ((103.58 +/- 5.48) cm, (16.60 +/- 2.21) kg, 65.80% +/- 18.54%, 65.10% +/- 18.92%, 0.46 +/- 0.53, 0.45 +/- 0.54, respectively) and the control group ((105.60 +/- 6.29) cm, (17.82 +/- 2.82) kg, 72.37% +/- 11.37%, 71.92% +/- 2.82%, 0.66 +/- 0.62, 0.66 +/- 0.52), the difference was significant (F values were 7.295, 8.034, 15.246, 10.745, 8.026, 10.864, respectively, P < 0.05).5-years of severe group, the value of height, the value of weight, the percentage of height, the percentage of weight, the SDS of height, the SDS of weight ((110.10 +/- 7.36) cm, (18.76 +/- 3.20) kg, 45.86% +/- 18.92%, 41.69% +/- 12.50%, -0.95 +/- 0.42, 0.23 +/- 0.34, respectively) were lower than the moderate group of the same age group ((117.76 +/- 6.35) cm, (21.63 +/- 2.75) kg, 61.81% +/- 20.75%, 61.79% +/- 18.92%, 0.36 +/- 0.62, 0.38 +/- 0.56) and the control group ((119.90 +/- 5.78) cm, (22.80 +/- 3.07) kg, 68.97% +/- 18.59%, 66.27% +/- 18.35%, 0.57 +/- 0.65, 0.48 +/- 0.63), the difference was significant (F values were 8.351, 7.864, 15.037, 13.921, 12.116, 11.725, respectively, P < 0.05).7 to 10 years-old of severe group, the value of height, the value of weight, the percentage of height, the percentage of weight, the SDS of height, the SDS of weight ((123.50 +/- 9.52) cm, (23.82 +/- 5.72) kg, 45.81% +/- 15.51%, 42.63% +/- 14.91%, -0.06 +/- 0.48, 0.02 +/- 0.61, respectively) were lower than the moderate group of the same age group ((129.1 +/- 8.41) cm, (26.70 +/- 5.72) kg, 66.84% +/- 16.09%, 64.07% +/- 18.58%, 0.48 +/- 0.46, 0.42 +/- 0.49) and the control group ((131.87 +/- 7.71) cm, (28.06 +/- 6.01) kg, 71.44% +/- 12.70%, 69.64% +/- 16.20%, 0.60 +/- 0.43, 0.60 +/- 0.51), the difference was significant(F values were 6.136, 6.678, 57.316, 37.893, 37.210, 34.152, respectively, P < 0.05). 3-, 5-, 7 to 10 year-old of moderate group, the value of height, the value of weight, the percentage of height, the percentage of weight, the SDS of height, the SDS of weight dropped compared to the control group of the same age, but no significant difference was found (t values were -2.008, -1.988, -1.810, -1.879, -1.713, -1.844, -1.904, 2.019, -1.605, -1.017, -1.411, -0.713, -1.881, -1.896, -1.746, -1.906, -1.523, 1.864, respectively, P > 0.05). CONCLUSION: The height and weight of children with severe asthma were lower than those of normal children or with moderate asthma. PMID- 21624240 TI - [Interventional effect of vitamin A supplementation on re-vaccination to hepatitis B virus among rural infants and young children in China]. AB - OBJECTIVE: The objective of this study was to observe the interventional effect of cod liver oil supplementation on re-vaccination to hepatitis B virus (HBV) among infants and young children. METHODS: All 7-36 months old infants and young children, who had been vaccinated with obligatory HBV vaccines routinely by the national technical and administrative procedures for HBV vaccination on children of China, were convened among villages in Linyi, Shandong province, from October 2008 to March 2009. After detection of serum anti-HBV, one hundred children with lower serum anti-HBV were picked out for the randomized, double blinded, placebo controlled vitamin A supplementation study. The children in the intervention group (50 subjects) took 0.5 g condensed cod liver oil (containing 25 000 IU vitamin A and 2500 IU vitamin D(2)) every 15 days for six times. The children in the control group (50 subjects) were given corn oil with same volume. All children were re-vaccinated at the 30th and the 60th day of the experiment. The serum samples were collected from each child at the 90th day of the experiment. Retinol concentration in serum samples was analyzed with HPLC method before and after the intervention. The levels of serum anti-HBs were detected by the electro chemi-luminescence immunoassay (ECLIA). RESULTS: Total 74 children finished the supplemental experiment and blood collection, 37 subjects in each group, respectively. After intervention, the serum retinol level in the experimental and control group were (404.1 +/- 123.1) and (240.8 +/- 92.8) ug/L (t = 6.441, P < 0.01), respectively. The serum anti-HBs levels in the experimental and control group were (2737.2 +/- 2492.6) and (1199.7 +/- 2141.6) U/L (t = 2.846, P < 0.01), respectively. The rate of weak or no-answer case in experimental and control groups was 0.00% (0/37) and 10.81% (4/37) (chi(2) = 4.229, P = 0.040), respectively. CONCLUSION: The results showed that vitamin A supplementation might enhance the re-vaccination reaction against HB vaccine in infants and young children. PMID- 21624241 TI - [The future of voice medicine in China]. PMID- 21624242 TI - [A few noteworthy points in clinical voice studies]. PMID- 21624243 TI - [Vocal fold augmentation by injection of autologous fascia and fat]. AB - OBJECTIVE: To evaluate the effect of combination of autologous fascia and fat injection into vocal fold for the treatment of patients with unilateral vocal fold paralysis and to observe the long-term effectiveness of this procedure. METHODS: A total of 26 unilateral vocal fold paralysis patients underwent vocal fold injection under general anesthesia, meanwhile, the mucosa of the injected point was sutured through laryngoscope under direct vision. There were 6 patients underwent autologous fat injection into vocal fold (group A), and 20 patients underwent autologous anterior rectus sheath fascia and fat injection (group B). Therapeutic efficacy were evaluated by videostroboscopy, voice-related parameters analysis and voice evaluation before and after treatment. Clinical analysis of this procedure was retrospectively performed in this serial of patients. RESULTS: All patients were followed up for 24 months. On the third day after operation, there was an acute inflammatory reaction induced by the graft. This reaction disappeared three months later. In all 20 cases, videolaryngostroboscopy showed significant improvement of the glottic closure, the improvement in acoustical parameters was statistically significant (P < 0.01). Perceptual evaluation of GRBAS scale showed significant improvement of phonatory function on G, B, A scale. The results remained stable 6 - 24 months after operation and were not changed by the length of follow-up. And in the 6 cases, videolaryngostroboscopy showed significant improvement of the glottic closure at 3 months compared with preoperative observation, a little spindle-shaped disclosure. The improvement in acoustical parameters was significant statistically at 3, 6 and 24 months (P < 0.05 or < 0.01), the voice quality decreased significantly at 6 and 24 months compared with 3 months (P < 0.05 or < 0.01). The significant differences were not observed between 6 and 24 months (P > 0.05). No complications were observed in all patients perioperatively or during the follow-up period. Voice-related parameters jitter, normalized noise energy and maximum phonation time showed significant differences between Group A and Group B on 24 months (P < 0.05 or < 0.01). CONCLUSION: The combination of autologous fascia and fat vocal fold injection is an effective procedure for the treatment of unilateral vocal fold paralysis, and the stable results can be achieved during the follow-up period for 24 months. PMID- 21624244 TI - [Analysis of the causes of 117 infants with persistent hoarseness]. AB - OBJECTIVE: To explore the causes of persistent hoarseness in infants. METHODS: One hundred and seventeen infants with persistent hoarseness treated in the department of otorhinolaryngology in Children's Hospital of Fudan University between June 2008 and July 2010 were retrospectively analyzed (all patients received antibiotic therapy for 2 weeks and the symptoms were not relieved after that). The patients were divided into three groups according to the age at first visit: 22 newborns, < 6 months old in 60 cases, < 12 months old in 35 cases. All patients had video laryngoscope examinations. Some of them received CT scan, cardiac ultrasonography and pathological examination in additional. The diagnosis was established by clinical history and imaging modalities, and the causes were analyzed subsequently. RESULTS: Among the 117 patients, 45 cases were vocal hypertrophy and hyperplasia (37.81%), 39 cases were vocal cord paralyses (32.78%), 7 cases were laryngeal hemangiomas (5.89%), 4 cases were laryngeal webs and cyst (3.36%), 2 cases were vocal cord polyps (1.68%), 2 cases were glottic incompetence (1.68%), 1 case was laryngeal papillomas(0.84%), 1 case was vocal code granulomas (0.84%), 1 case was glottis restricted by neck lymphangioma (0.84%); 4 cases were undetermined and 13 cases were no abnormalities. The percentage of patients with congenital heart diseases (19 cases) in vocal cord paralysis was 48.72%. The proportion of vocal cord paralysis in younger group was higher than that in elder one, their percentage were 50.00%, 36.67% and 17.14% respectively (chi(2) = 7.18, P < 0.05). CONCLUSIONS: A variety of causes can lead to persistent hoarseness in infants. The majority of them are vocal hypertrophy and hyperplasia, followed by vocal cord paralyze. Vocal cord paralysis is more common in younger infants than in elder ones, and the main causes are post cardiac surgery and congenital heart disease. PMID- 21624245 TI - [Effectiveness of voice training for professional voice users with voice disorders]. AB - OBJECTIVE: To explore the effectiveness of voice training on professional voice users with voice disorders. METHODS: Thirty one professional voice users suffering from voice symptoms without organic disorder were treated with a four week voice training program, including vocal hygiene education, respiratory training and resonant therapy. Voice handicap index, maximum phonation time, noise to harmonic ratio, jitter and shimmer were compared before and after the training. RESULTS: The voice handicap index (x(-) +/- s) decreased from 33.7 +/- 19.2 to 18.8 +/- 18.4 after the training and the difference was significant (t = 6.14, P < 0.05). Maximum phonation time (x(-) +/- s) increased significantly from (15.5 +/- 5.8) s to (18.6 +/- 6.0) s (t = -3.43, P < 0.05). jitter (M [P(25); P(75)]) decreased significantly after the training from 0.42% [0.36%; 0.62%] to 0.35% [0.29%; 0.47%] (Z = -2.51, P < 0.05), while there was no significant change in shimmer and noise to harmonic ratio. CONCLUSION: Voice training can effectively reduce the vocal symptoms in professional voice users, decrease the handicap degree and increase the respiratory support during phonation, thus improve the voice quality. PMID- 21624246 TI - [Experimental study on adipose-derived stem cells combined with acellular dermal matrix particles for vocal fold injection]. AB - OBJECTIVE: To explore the feasibility of adipose-derived stem cells (ASC) combined with micronized acellular dermal matrix (MADM) for vocal cord injection. METHODS: The adipose-deprived stem cells were harvested from rabbit adipose tissue in vitro. The 3rd generation of ASC was labeled with DiI (1,1-dioctadecyl 3,3,3,3-tetramethylindocarbocyanine perchlorate) and cultured with MADM to form a complex. The adhesion of ASC to MADM was observed by fluorescence microscope and electron microscope. The proliferation of ASC on MADM was evaluated by 3-(4,5 dimethylthiazol-2-yl)-5-(3-carboxy methoxyphenyl)-2-(4-sulfonyl)-2H-tetrazolium, inner salt (MTS). Three days after the culture, the complex was mixed with appropriate amount of collagen, and then injected into the unilateral vocal cord of the rabbit. The animals were sacrificed 2, 4, 8 weeks after injection, the survival time and distribution of ASC in vocal fold were tested, and the responses of vocal cord to ASC-MADM and the degradation of MADM were observed. RESULTS: The ASC adhered to MADM and grew well (P < 0.05 or < 0.01), showing good compatibility with MADM in vocal cord tissue. The complex of ASC-MADM could be injected into the rabbit vocal cords, while no adverse reactions was observed in the vocal cord by endoscope, frozen section and HE staining. ASC could survive for 8 weeks in vocal cords, and no inflammatory cell infiltration was observed. CONCLUSIONS: MADM is an ideal scaffold material and shows perfect compatibility with ASC which can adhere and proliferate well on it. The complex of ASC-MADM can be injected into the vocal cord and can survive. There is no adverse reaction in vocal cords. PMID- 21624247 TI - [Preliminary research on the regeneration of injured rabbit vocal folds after the transplantation of human amniotic epithelial cells]. AB - OBJECTIVE: To investigate the survival, growth and distribution of human amniotic epithelial cells (hAEC) after injected into injured rabbit vocal folds, in addition, to assess the ability of hAEC to affect the components of lamina propria extracellular matrix (ECM) and prevent vocal fold scarring. METHODS: hAEC were isolated from human amnion and marked by Lenti-EGFP. Fifteen New Zealand rabbits were used for this experiment. EGFP-hAEC was injected into the left injured vocal folds in thirteen rabbits, and the contralateral thirteen vocal folds experienced an injured procedure only ("injured untreated control"), and four vocal folds were left as untreated controls. The survival, distribution, differentiation potential and secretion function of hAEC were examined by immunofluorescence method. HE staining and immunohistochemical staining were performed for the evaluation of collagen and fibronectin respectively. RESULTS: hAEC showed a cobblestone-like growth. After implanted into the injured vocal folds, hAEC could survive in vocal fold lamina propria for 2 months. The immunofluorescence analysis showed the evidence of hAEC differentiation into muscle cells as well as secretion the ECM protein. Three months postoperatively, the density of collagen was higher in the injured untreated control folds than that in the injured vocal folds injected with hAEC and the untreated controls. Besides, the content of fibronectin in the injured untreated control group was significantly increased. CONCLUSIONS: hAEC survived in the vocal folds lamina propria, and had the potentiality to differentiate into vocal folds tissue and secret some ECM components. The histological improvement caused by the injected cells demonstrate that hAEC had the ability to promote the repairment and regeneration of injured vocal folds. PMID- 21624248 TI - [Activity of bilateral posterior cricoarytenoid muscle satellite cell after denervation or reinnervation with ansa in dogs]. AB - OBJECTIVE: To investigate the activity of bilateral posterior cricoarytenoid muscle satellite cell after denervation or reinnervation with ansa cervicalis. METHODS: Twenty four dogs were randomly divided into 3 groups. The bilateral laryngeal recurrent nerves were cut in group one in all dogs. The bilateral laryngeal recurrent nerves were anastomosed with ansa cervicalis after incision in group two in all dogs. The dogs in group three were used as control. Nine weeks after surgery, the electromyography was used to test the regeneration of the nerve. The posterior cricoarytenoid muscles biopsy were collected. The expression of mRNA of Myogenin, Myf5, and Pax7 was assayed by realtime RT-PCR after total RNA isolation. RESULTS: Two dogs died after surgery in incision and anastomose group. The electromyography suggested that the RLN of all dogs had denervated in the incision group and had reinnervated in the anastomose group after 9 weeks. Myogenin mRNA from RLN incision dogs PCA muscles had greater expression versus controls (Z = 1.42, P < 0.01) or anastomosed dogs (Z = 1.38, P < 0.01). Myf5 mRNA expression from RLN incision dogs PCA muscles had significant increase versus control dogs (Z = 1.66, P < 0.01) or anastomosed dogs (Z = 1.69, P < 0.01). Pax7 mRNA expression from RNL incision dogs had significant increase compared with control (Z = 1.66, P < 0.01) or anastomosed animals (Z = 1.42, P < 0.05). There was no significant difference in Myogenin (Z = 1.34, P > 0.05), Myf5 (Z = 0.54, P > 0.05) and Pax (Z = 0.54, P > 0.05) mRNA expression between controls and anastomosed animals. CONCLUSIONS: The bilateral denervation of RLN cause significantly increasing in dog PCA muscle satellite cell proliferation and differentiation. The bilateral reinnervation of RLN cause PCA muscle satellite cell come back nonproliferative, quiescent state in dog. PMID- 21624249 TI - [Expression and significance of matrix metalloproteinase and tissue inhibitor of metalloproteinase in nasal cavity squamous cell carcinoma]. AB - OBJECTIVE: To study the expression and significance of matrix metalloproteinase (MMP) 2, MMP-9, tissue inhibitor of metalloproteinase (TIMP)1 and TIMP-2 in nasal cavity squamous cell carcinoma and their signification. METHODS: Fifty cases of epithelial carcinoma tissue and 50 cases of normal nasal tissue were detected for MMP-2, MMP-9, TIMP-1, TIMP-2 by immunohistochemistry technique (S-P), and their relationship between the expression of MMP-2, MMP-9, TIMP-1, TIMP-2 and some clinical symptoms were analyzed. The SPSS 12.0 software was used to analyze the data. RESULTS: The positive ratio of expression of MMP-2 in 50 cases of epithelial carcinoma was 52.0% (26/50), which was significantly higher (chi(2) = 6.00, P < 0. 05) than those [28.0% (14/50)] in the normal nasal tissue. The positive ratio of expression of MMP-9 in 50 cases of epithelial carcinoma was 58.0% (29/50), which was significantly higher (chi(2) = 12.8, P < 0.05) than those [10.0% (5/50)] in the normal nasal tissue. The positive ratio of expression of TIMP-1 in two groups was 74.0% (37/50), 56.0% (28/50) respectively. There was no difference between two groups (chi(2) = 0.51, P > 0.05). The positive ratio of expression of TIMP-2 in two groups was 26.0% (13/50), 20% (10/50) respectively. There was no difference between two groups (chi(2) = 3.35, P > 0.05). CONCLUSIONS: There was a close relationship between pathogenesis and development of nasal epithelial carcinoma and the expression of MMP-2, MMP-9 in the epithelial carcinoma tissues. Both MMP and TIMP, especially the unbalance of MMP and TIMP, have prognostic value in nasal cavity squamous cell carcinoma. PMID- 21624250 TI - [Correlation between patient-based questionnaires and computer tomography staging in chronic rhinosinusitis]. AB - OBJECTIVE: To investigate the relationship between the patient-based questionnaires and the computed tomography (CT) staging in patients with chronic rhinosinusitis (CRS). METHODS: Quantitative data of 121 preoperative recruits with CRS were collected by using the Lund-Mackay CT staging system, a visual analogue scale (VAS), sino-nasal outcome test-20 (SNOT-20), and the medical outcome study short-form 36 items (SF-36). The patients were classified into several subgroups according to whether CRS was associated with nasal polyps (NP) or not, sex, duration of disease, and educational background. Correlation between the patient-based questionnaires and the CT staging were analyzed in the total cohort patients and subgroups. RESULTS: In the total cohort patients, there were significant correlations between SNOT-20 and SF-36 (r = -0.561, P < 0.01), SNOT 20 and VAS (r = 0.743, P < 0.01), and SF-36 and VAS (r = -0.504, P < 0.01), however, the CT staging did not correlate with the patient-based questionnaires (P > 0.05). Significant but weak correlations were found between the CT staging and the patient-based questionnaires in the CRS with NP subgroup (CT vs SNOT-20, r = 0.318, P = 0.005; CT vs SF-36, r = -0.358, P = 0.002; CT vs VAS, r = 0.358, P = 0.002). Compared between CRS with NP and without NP subgroup, there were statistic differences on the Lund-Mackay CT stage and the SNOT-20 and VAS scores (t value was 3.249, -2.409, -2.957, respectively, all P < 0.05). CONCLUSIONS: The patient-based questionnaires correlate well with each other, but CT staging correlated significantly but weakly with the patient-based questionnaires only in the CRS with NP subgroup. Nasal polyps do not appear to be responsible for the adverse effects of CRS on quality of life. PMID- 21624251 TI - [DNA methylation and histone modification relate to RASSF1A gene deletion in laryngeal carcinoma tissues]. AB - OBJECTIVE: To investigate the relationship between RASSF1A gene expression and DNA methylation or histone modification in laryngeal carcinoma tissues. METHODS: Chromatin immunoprecipitation (ChIP), methylation specific polymerase chain reaction (MSP) and realtime quantitative reverse transcription polymerase chain reaction (realtime RT-PCR) were used to analyze RASSF1A gene promoter region histone H3 lysine 9 methylation, H3 lysine 4 methylation, H3 lysine 9 acetylation, DNA methylation, and RASSF1A gene expression in laryngeal carcinoma tissue of 50 cases. RESULTS: DNA methylation rate of gene RASSF1A was 62% in 50 cases of laryngeal carcinoma, but no DNA methylation was found in normal control group, with a significant difference (chi(2) = 15.381, P < 0.05). DNA methylation had no correlation with age, gender, differentiation degree, T stage, pathological type and lymph node metastasis (P > 0.05). The affection of DNA methylation group was more than unmethylation group to expression of gene RASSF1A (t = -3.108, P < 0.01). There was positive correlation between RASSF1A deletion and gene hypermethylation or between H3 lysine 9 methylation of RASSF1A gene promoter and DNA methylation in laryngeal carcinoma tissue(r = 0.816, P < 0.05), but there was negative correlation between H3 lysine 4 methylation of RASSF1A gene promoter and DNA methylation (r = -0.837, P < 0.05) and no correlation between H3 lysine 9 acetylation and DNA methylation (r = -0.383, P > 0.05). CONCLUSIONS: Laryngeal tumor suppressor gene RASSF1A promoter methylation is a key factor down-regulating the gene expression, and histone modifications also plays an important role in tumor development. PMID- 21624252 TI - [Isolation, culture and growth characteristics of human muscle stem cells]. AB - OBJECTIVE: To establish the methods for purification, culture, and identification of adult human skeletal muscle stem cells in vitro and to explore the biological properties of the cells. METHODS: Muscle stem cells were obtained by reformed enzymatic digestion of muscle tissue from the consenting donors and cultured in serum-free medium. The morphology was inspected by an inverted phase contrast microscope. Phenotypic characteristics of the cells and expression of cell specific markers were determined using reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry. The growth of single cells in suspension culture was observed and recorded continuously. The cells were analyzed for their multi-lineage differentiation potential into osteoblastic, adipocyte, and smooth muscle cell lineages. RESULTS: Primary cultured human skeletal muscle stem cells proliferated and formed the big spheres when cultured with serum free medium. Immunofluorescence staining displayed Pax7 and ALDH1 positive expression in the cell spheres. Furthermore, Myod and Desmin showed positive expression in the monolayer cells derived from the spheres. The gene expressions of Oct3/4, Nanog, Sox2 and Pax7 in the cells were determined by RT PCR. The cell clones formed from single cells grew well. In addition, they were capable of spontaneous differentiation into myotubes in differentiation medium and into other mesodermal cell lineages in induction medium. CONCLUSIONS: Human muscle stem cells with properties of self-renewal capacity and multi differentiation could be successfully isolated and expanded in vitro. PMID- 21624253 TI - [Origin of acoustically evoked short latency negative response in guinea pigs]. AB - OBJECTIVE: To establish a model of acoustically evoked short latency negative response (ASNR) in guinea pigs, a model of profound hearing loss with normal saccular functions, and verify the correlation between ASNR and vestibular evoked myogenic potential (VEMP). METHODS: Thirty-two healthy guinea pigs were employed in the experiment, which were randomly divided into control group (16 subjects) and deafened group (16 subjects). Each animal experienced auditory and vestibular tests including auditory brainstem response (ABR), VEMP and caloric test. A quick treatment was employed for deafened group consisting of a subcutaneous injection of kanamycin at a dose of 400 mg/kg followed by a jugular vein injection of ethacrynic acid at a dose of 40 mg/kg one hour later. The animals were received ABR, VEMP and caloric test 7 - 10 days following the drug administration. The deafened group was further divided into ASNR group and non-ASNR group, based on the presence of ASNR. RESULTS: In deafened group, five subjects died postoperatively, 11 subjects (22 ears) provided full data, ASNR was elicited in eight ears (36.4%), the threshold was 120 - 130 dB SPL with mean of (124.4 +/- 4.96) dB SPL. Its latency range was 1.75 - 2.60 ms with mean of (2.15 +/- 0.27) ms. The mean latency of threshold was (2.34 +/- 0.18) ms. All eight ASNR ears presented with VEMP. The VEMP threshold, positive and negative potential latencies proved no statistical difference (P > 0.05) between ASNR group and control group. Significant difference was detected between the VEMP presence of ASNR group and non-ASNR group (P = 0.002). There was no statistically significant correlation between VEMP and caloric test neither between ASNR and caloric test in deafened group. CONCLUSIONS: This study evoked ASNR in an ototoxicity guinea pig model which has profound hearing loss with normal saccular functions. The presence of ASNR correlated with VEMP, however, not correlated with caloric test, suggesting that ASNR and VEMP are both originated from the saccule. PMID- 21624254 TI - [Role and mechanism of the expression of matrix metalloproteinase 9 and tumor necrosis factor alpha in upper and lower respiratory tract inflammation in rats]. AB - OBJECTIVE: To investigate the role and mechanism of matrix metalloproteinase 9 (MMP-9) and tumor necrosis factor alpha (TNF-alpha) in the pathogenesis of allergic rhinitis (AR) and asthma(AS). METHODS: The rat models of AR and AS were made by injecting ovalbumin. The infiltration of eosinophils and mast cells were detected by hematoxylin-eosin (HE) staining and toluidine blue staining respectively, and the expression of MMP-9 and TNF-alpha in nasal mucosa and lung tissue were examined by immunohistochemical staining (SP method). The relationship of their expression with upper and lower respiratory tract inflammation was analyzed. SPSS 13.0 software was used to analyze the data. RESULTS: The numbers of MMP-9 positive inflammatory cells in nasal mucosa and lung tissue of AR were (154.8 +/- 12.0) and (124.0 +/- 8.2), (43.2 +/- 7.6) and (34.5 +/- 5.0) in the control groups, the difference was significant (t value were 24.260, 29.525 respectively, all P < 0.05). The numbers of MMP-9 positive inflammatory cells in nasal mucosa and lung tissue of AS were (149.9 +/- 11.7) and (120.1 +/- 7.3), (48.6 +/- 7.6) and (39.1 +/- 5.2) in control groups, the difference was significant (t value were 22.929 and 28.530 respectively, all P < 0.05). The numbers of TNF-alpha positive inflammatory cells in nasal mucosa and lung tissue of AR were (188.8 +/- 17.0), and (134.8 +/- 7.9), (57.6 +/- 23.3) and (40.3 +/- 8.2) in control groups, the difference was significant (t value were 13.836 and 26.220, all P < 0.05). The numbers of TNF-alpha positive inflammatory cells in nasal mucosa and lung tissue of AS were (179.2 +/- 15.4) and (153.5 +/- 10.1), (70.5 +/- 33.1) and (33.8 +/- 14.0) in control groups, the difference was significant (t value were 9.412 and 21.858, all P < 0.05). There was a correlation between the expression of MMP-9 and TNF-alpha in nasal mucosa and lung tissue of AR (r values were 0.893 and 0.700 respectively, P values were 0.001 and 0.024, respectively) and AS (r values were 0.692 and 0.644 respectively, P values were 0.027 and 0.044 respectively) groups. CONCLUSIONS: The inflammation is similar between AR and AS. The MMP-9 and TNF-alpha may play an important role in the pathogenesis of upper and lower respiratory tract inflammation. PMID- 21624255 TI - [Biomechanical study on decellularized laryngeal scaffold in dogs]. AB - OBJECTIVE: To evaluate the biomechanical characteristics of the decellularized laryngeal scaffold. METHODS: Ten Chinese adult dogs were randomly divided into two groups: perfusion group (n = 5) and control group (n = 5). The acellular larynx scaffold was obtained from dogs through cranial thyroid arteries perfusion with detergents. Comparative examinations were performed by the macroscopic view, histological view (hematoxylin and eosin stain, Alcian blue stain and Masson stain), scanning electron microscope (SEM) and biomechanical properties between perfusion group and control group. RESULTS: Macroscopic view showed that the decellularized laryngeal scaffold appeared pale asphyxia. HE stain indicated that there were little acellular traces of muscle and mucosa. Alcian blue stain, Masson stain and scanning electron microscope (SEM) suggested that there were no obvious changes about glycosaminoglycan and collagen. The compressive modulus of thyroid cartilage was (1.06 +/- 0.07) MPa (x(-) +/- s) in experimental groups and (1.15 +/- 0.11) MPa in control group, showing no significant difference (t = 1.424, P > 0.05), neither in compressive modulus of annular cartilage (1.68 +/- 0.11) MPa in experimental groups and (1.67 +/- 0.09) MPa in control group (t = 0.185, P > 0.05). The tensile strength of thyroid cartilage between experimental (5.74 +/- 0.88) MPa and control groups (6.18 +/- 1.33) MPa did not have the statistical significance (t = 0.627, P > 0.05). CONCLUSION: These results indicate that perfusion method can construct a perfect biomechanical acellular larynx scaffold which could be a better selection for laryngeal reconstruction with tissue engineering method. PMID- 21624256 TI - [Coblation treatment for laryngeal papilloma in adult]. AB - OBJECTIVE: To evaluate the effect and feasibility of coblation treatment for laryngeal papilloma in adult. METHODS: A total of 18 patients with laryngeal papilloma treated by coblation from April 2008 to June 2010 was retrospectively analysed. There were 4 cases of multiple laryngeal papilloma and 14 cases of single laryngeal papilloma. All of these patients were treated with CoblatorTM Reflex #7070 under general anesthesia without tracheotomy. RESULTS: The volumes of blood loss during surgery varied from 1 ml to 10 ml, 2 ml on average. There was no postoperative bleeding or other complications. Follow-ups ranged from 6 months to 33 months after surgery, median 18 months. Recurrence occurred 4 months and 8 months after surgery in two patients. They were treated with coblation for the second time and followed up for 4 months and 12 months respectively. Recurrence occurred again in one patient. CONCLUSION: Coblation under suspension laryngoscope for laryngeal papilloma in adult was an ideal way due to less bleeding and less damage. PMID- 21624257 TI - [Characteristic ultrasonographic features of the encapsulated papillary thyroid carcinoma]. AB - OBJECTIVE: To evaluate the ultrasound (US) features of the encapsulated papillary thyroid carcinoma (EPTC). METHODS: Based on ultrasonographic features including shape, size, border, echogenicity, hypoechoic halo and microcalcification, 33 cases of EPTC were classified into two groups: 21 cases in irregular shape group and 12 cases in spherical or oval shape group. RESULTS: EPTC in the irregular shape group showed some ultrasonographic features including jagged border, irregular tumor shape and marked hypoechogenicity, while the ultrasonographic features of EPTC in the spherical or oval shape group included smooth border, regular shape, isoechogenicity and hypoechoic halo. Hypoechoic halo and isoechogenicity were found more frequently in EPTC of spherical or oval shape group than those in EPTC of irregular shape group. The size of EPTC in the spherical or oval group was commonly larger than that of EPTC in the irregular shape group. CONCLUSIONS: The findings indicate that EPTC have some ultrasonographic features similar to benign follicular thyroid tumors. PMID- 21624258 TI - [A Jarisch-Herxheimer reaction misdiagnosed as pneumonia after an operation for laryngeal papillary lymphoma]. PMID- 21624259 TI - [Neurofibromatosis I style in the salivary gland and spheno-maxillaris fossa]. PMID- 21624260 TI - [A case of foreign body in esophagus of a child]. PMID- 21624261 TI - [Neurofibroma in nasal cavity]. PMID- 21624262 TI - [Nasal vestibule cutaneous metastase of cancer: a case report]. PMID- 21624263 TI - [Current practice in injection laryngoplasty]. PMID- 21624264 TI - [Temporary balloon occlusion test in the assessment of cerebral collateral circulation]. PMID- 21624265 TI - [Korean doctor Che-ma Yi and his Sasang Constitutional Medicine]. AB - Tongui suse powon, written by Chi-ma Yi (1837 - 1900), divided people into four types according to their constitution and disposition-Taiyang, Taiyin, Shaoyin and Shaoyang. He discussed the appearance, viscera, temperament and disease each type of people probably have. The formation of Sasang Consititutional Medicine is influenced by Sasang Theory in Yi Jing and 25-mode Personality Theory in Ling Shu and has nothing to do with Huangji Jingshishu written by SHAO Yong. PMID- 21624266 TI - [Hanke Zhiyan-a vivid reflection of medical exchanges history between Japan and Korea]. AB - Hanke Zhiyan (written by Dooyo Higuchi and published in 1749), held in the Fujikawa collection of Kyoto University Library, recorded in detail the process by which Higuchi was assigned by the government to heal Korean envoys. The name, age, symptoms, diagnosis and medicine of 14 patients were described in detail in this book. It could be known from the recording that Korean envoys suffered from a variety of diseases and Dooyo Higuchi (a Traditional Chinese Medicine doctor of Japan), with a solid academic foundation and plenty of experience, could give specific diagnoses and flexible therapies. Hanke Zhiyan described full and accurate medical exchanges between Japan and Korea, and was a vivid reflection of medical exchanges in the history of the two countries. PMID- 21624267 TI - [Statistical analysis of existing ancient medical works in Japan and Korea and the development value]. AB - The category, number, date of writing, circulated copies and library collection situation of 12275 Japanese medical works before 1912 and 416 Korean medical works before 1910 were analyzed. Most of these medical books are clinical books, which reflected the emphasis on clinical practice and conclusions in the two countries. Japanese medical books were mainly completed between the 17 - 19th century and nearly all of them are conserved as copies, while in Korea the number is 39.6% and we don't know the time that more than half of Korean medical books were completed. The existing books in Japan are collected in more than 500 libraries, especially in 19 public and private libraries., such as Kyoto University Library, Tokyo University Library, Japan Library of Congress. In Korea, most of the books are conserved in Kyujanggak Institute for Korean Studies in Seoul National University. It is necessary for scholars of China, Japan and Korea to work together to study and utilize the ancient medical books of the three countries and to make them play a more important role in the development of Eastern traditional medicine. PMID- 21624268 TI - [Status and prospect of protection of intangible cultural heritage-traditional medicine in the international social community]. AB - In the 1970s, the World Health Organization (WHO) began to focus on traditional medicine and realized the cultural foundation of it. The United Nations Educational, Scientific and Cultural Organization (UNESCO) began the work of standardization of intangible cultural heritage in 1973 and in 2003 the Convention for the Safeguarding of Intangible Cultural Heritage was approved. The categories of intangible cultural heritage kept on increasing and the adoption of traditional medicine began in 2003. Till now, two traditional medical items have been included in The Representative List of the Intangible Cultural Heritage of Humanity. Now intangible cultural heritage has been emphasized and supported by many countries rich in cultural resources. The number of member states and items in the list increased rapidly. The aim of The Convention for the Safeguarding of Intangible Cultural Heritage, protecting the cultural foundation of traditional medicine and promoting the inheritance and revitalization of core cultural conception in traditional medicine, is a new way for sustainable development of traditional medicine in the future. PMID- 21624269 TI - [Demotion and promotion of CAI Jing and the Medicine School's establishment and abolition three times in the North Song dynasty]. AB - CAI Jing was appointed the prime minister in the Chongning period of Song Hui tsung. The Medical School was moved to the Imperial College from Taichang Temple. It used a 3-year education system and divided graduates into three grades. The preferential policies promised top students as 8 or 9-rank official position, which attracted a lot of intellectuals into the field of medicine. CAI Jing was demoted three times respectively in the fifth year of the Chongning period, the third year of the Daguan period and the second year of the Zhenghe period, and was promoted again after each demotion. Influenced by changes of CAI Jing's position and relative policies, the Medical School was also established and abolished three times. PMID- 21624270 TI - [Analysis of governmental administration on medical advertisements in modern China]. AB - The huge medical market in modern China brought about prosperity and then an uncontrolled medical advertising industry. In order to standardize the market, the health institution resorted to laws and regulations. Shanghai Municipal Health Bureau began the work in 1928 and Shanghai Provisional Rules of Banning Obscene Drug Advertising was issued in 1929. In 1930, Provisional Rules of Chinese and Western Medicine Advertising of the Beiping Health Bureau was carried out and then Rules of Chinese and Western Medicine Advertising of Beiping in 1935. From then on, various parts of the country kept on developing and perfecting their own rules on the basis of Rules of Chinese and Western Medicine Advertising of Beiping and the local situation. Among the rules, Provisional Rules of Chinese and Western Medicine Advertising of Shanghai (1936), Rules of Administrating and Correcting Chinese and Western Medicine Advertising of Beiping (1946) and Rules of Medicine Advertising of Shanghai Municipal Health Bureau (1947) were representatives. Those rules embodied the modern government's principle on administrating advertising by legal means and restraining excess illegal medicine advertising. There were also some weak points in the supervision of medicine advertising, such as sluggish supervision, ineffective supervision and light punishment. PMID- 21624271 TI - [Early Activities of China Medical Board's in China]. AB - China Medical Board (abbreviated as CMB), originated from Rockefeller Foundation, gives support specifically to the medical development and health promotion in China and other underdeveloped areas in Asia. At its beginning, CMB's vision is to lay a solid and systematic foundation with high standard for the long-term development of medicine in China. With this vision, CMB of Rockefeller Foundation built up Peking Union Medical College (PUMC), the best one at that time in Asia and the world.. Besides this main task, from 1914 to 1919, CMB also devoted its effort and funding to the development of other missionary medical colleges in China, sponsored missionary hospitals, provided fellowship, scholarship and grants to medical professionals and staff in China, which greatly enhanced the development of not only PUMC but also the advancement of medicine and promotion of health in China. PMID- 21624272 TI - [Human factors caused the third plague epidemic in Harbin]. AB - The third plague epidemic in Harbin broke out in 1946 and ended in 1954. Different from the first two plague epidemics (imported), the third prevalence was both imported and idiopathic infectious disease which was caused by human factors. Japanese troops set forbidden zones to build a biological weapon center, which destroyed the natural environment and offered a good growth condition for Citellus Undulatus. In 1945, on the eve of surrender, the Japanese blew up the Unit 731 germ factory located in a bungalow district, which caused diffusion of infected plague fleas. Mice of the district were infected and a man-made plague focus was created. During the prevalence of the third plague, prevention departments at all levels took a series of actions and with people's efforts, the plague was effectively controlled. PMID- 21624273 TI - [Study of the Korean medical book-Suwen Dayao]. AB - The Korean, Gyoo-joon Lee (1855 - 1923, also named Shigu Shanren), was born in Yingri County, Qingbeidao and was familiar with literature and history and Chinese medicine. In 1906, he completed the book Suwen Dayao, selecting 25 chapters from Suwen and deleted WAND Bing's notes but made some own notes with his excellent ideas. The book is in a block-printed edition (in 1906, few and held in Xinglin Academy) and a hand copy edition (by his disciple Won-se Lee in 1921). Gyoo-joon Lee was familiar with Suwen and was good at clinical practice and his medical theory could be seen in his papers-Suwen Fushuo and Baibing Zongkuo in Chapter 4 of Suwen Dayao which embodied the close combination of clinical practice and medical theory. Baibing Zongkuo was written in seven character regulated verses which reflected Gyoo-joon Lee's ability of reading and writing in Chinese. There are many outstanding ideas in Suwen Dayao. The Fangzhi Bafa written by his disciple Jong-sun Lee was attached to the end of the book, which are valuable for the combination of the theory in Suwen, decoctions in Shanghan Lun and current decoctions. Suwen Dayao expanded the influence of Huangdi Neijing in Korea and fostered a lot of talent. PMID- 21624274 TI - [The Korean and Japanese editions of the Korean medical book-Dongyi Baojian]. AB - From 1613, Dongyi Baojian (written by Huh Joon) has been published in more than 30 editions in Korea, China and Japan. There are 13 kinds of editions in Korea movable type edition in the Kuichou year; block printed Lingying edition in the Jihai year; the first Lingying edition in Jiaxu year; the edited Lingying edition in the Jiaxu year and the Wanying reprint edition in the Jiaxu year. There are 2 kinds of editions in Japan-the first edition in the Xiangbao period and the reprint edition in the Kuanzheng period. PMID- 21624275 TI - [Study of dates of birth and death of WANG Tao]. AB - The dates of birth and death of WANG Tao, a famous doctor in Tang dynasty, are not found in literature and scholars have different points upon this subject. It could be sure that WANG Tao was appointed mayor of Huayuan county in the ninth year of Kaiyuan period (721) and in December of the fourteenth year of Tianbao period (756) he the revolt in Hejian. The recording after 756 could not be found in literature. According to the official system in Tang dynasty that officers retired at their age of 70, it could be sure that WANG Tao was less than 70 year old in 756. Considering his age when he was the county mayor in 721, it could be speculated that WANG Tao was born during the third year of Chuigong period (687) to the second year of Changshou period (693), and his death date should be some date after 756. But it couldn't be validated. PMID- 21624276 TI - [Study on teachers and disciples of DOU Hanqing]. AB - DOU Hanqing, a doctor in the Jin and Yuan dynasty, was good at acupuncture. In 1218, the 23-year-old DOU Hanqing married a daughter of medical family in the Qingliuhe area. DOU Hanqing began to learn medicine from the family. From then on, he learned acupuncture from SONG Zihua, LI Hao and his son (famous doctors in Shandong), XIE Xianzi, QIU Changsheng and others, and his achievements were great. DOU Hanqing was not only good at acupuncture but also was pleased to teach students. WANG Renzheng and his son, ZHENG Yu, ZHU Yanhui, XU Kan, LIU Zhizhong and LUO Tianyi were all his students and a few his hometown people also learned from him. PMID- 21624277 TI - [The first nursing school of Guangxi in modern times]. AB - In 1903, the Sida Anniversary Hospital was established in Wuzhou, Guangxi, by the American Baptist Convention. In 1909, the head nurse Leonora Swrlett was sent to Wuzhou from USA to take charge of nursing work. Considering the lack of nursing staff, Leonora Swrlett established the Primary Vocational School of Nursing Affiliated to Guangxi Sida Hospital. As the beginning of women's medical education, the establishment of the Primary Vocational School of Nursing enriched the variety of education, and introduced advanced education modes and systems, which were of great value for establishing nursing schools and had huge influence on modernization of women's education in Guangxi. PMID- 21624278 TI - [ZHENG Guanying's ways of the health protection from Zhongwai Weisheng Yaozhi]. AB - ZHENG Guanying (1842 - 1922) was a famous industrialist and educator of modern times. He is interested in medicine and dabbled in health preserving of traditional Chinese medicine and health care of western medicine which could be reflected in his books Zhongwai Weisheng Yaozhi. His opinions on mental regulation, healthy diet and daily life could be seen from this book. PMID- 21624279 TI - [Study of pediatric allergic diseases by using an interdisciplinary path]. PMID- 21624280 TI - [Historical development and challenges of the child health care in China]. PMID- 21624281 TI - [Diagnostic capacity of skin prick test in egg and cow's milk allergic infants]. AB - OBJECTIVE: Mean diameter is the most common used parameter for wheal response assessment after skin prick test. This study aimed to investigate the diagnostic capacity of mean diameter according to the outcome of oral food challenge, and to determine the cut-off points that could render food challenges unnecessary. METHOD: Data of 173 children referred to the Division of Primary Child Health Care for the evaluation of suspected food allergy were prospectively studied. All children underwent skin prick test and open food challenge to the relevant food(s) in clinic. The mean wheal diameter of skin prick test was measured, and open food challenge was performed to confirm food allergy. The SPSS software package version 13.0 for windows (SPSS, Chicago, IL, USA) was used for all statistical analysis. Open food challenge was taken as the gold standard for diagnosis. Diagnostic capacity of skin prick test, including the sensitivity, specificity, positive predictive value, negative predictive value, was calculated by cross-table. In addition, receiver operating characteristic curve (ROC) was plotted and area under the curve (AUC) was calculated to quantify the accuracy of the parameter. RESULT: For the 173 children, 271 open food challenges were performed with egg white, egg yolk and cow's milk, In which 123 were positive, 99 children were diagnosed as food allergy. Cutaneous symptoms (87.0%) were most common, followed by gastrointestinal symptoms (9.8%). The AUC of mean diameter was 0.794 for egg white, 0.804 for egg yolk and 0.904 for cow's milk. The sensitivity of skin prick test with a cut-off value of >= 3 mm was ranged from 71% to 87%, while the specificity was between 31% and 57%. The authors also defined food specific skin prick test mean diameters that were 100% diagnostic for allergy to egg white (>= 8.5 mm), egg yolk (>= 5.5 mm), cow's milk (>= 5.5 mm). CONCLUSION: Predictive decision points for a positive outcome of food challenges can be calculated for egg and cow's milk using mean diameter. It may help to simplify the diagnostic procedure of food allergy. PMID- 21624282 TI - [Follow-up study of school-age children's sleep quality in Shanghai]. AB - OBJECTIVE: China has undergone massive socioeconomic change during the past several years, and its impact on children's sleep is still unrecognized. Shanghai, as one of typical economically fast-developing cities, was chosen as observational city in this study, which was designed to explore trends in sleep quality in Shanghai school-aged children and related high risk factors on sleep quality. METHOD: Totally 884 fifth grade school-aged students were sampled by stratified cluster random sampling method from 10 primary schools of Shanghai in 2005, then four years later in 2009, 2161 same grade students were sampled from the same schools. Chinese version of Children's Sleep Habits Questionnaire (CSHQ) was used to evaluate 8 sleep quality parameters among those children, and high risk factors on school-aged children's sleep quality were investigated as well. RESULT: The prevalence of poor sleep quality decreased from 29.2% in 2005 to 27.9% in 2009, and among 8 sleep quality parameters, bedtime resistance decreased from 33.1% to 28.7%, sleep anxiety from 50.6% to 39.8%, night waking from 25.2% to 21.5%, and parasomnia from 51.2% to 45.8%. The factors, such as heavier homework burden, longer daily computer use, bright light during sleep, cosleeping, existence of chronic disease and irregular sleep habits of parents, were associated with poor sleep quality of school-aged children after adjusting for children's age, gender, and family social-economic status. CONCLUSION: Part of sleep quality parameters improved during the past 4 years, but current situation is still tough with more than 1/4 poor sleep quality children. Helping children to develop good sleep hygiene as well as educating parents how to shape children's regular sleep habits might be effective methods to improve children's sleep quality. PMID- 21624283 TI - [Variation of maternal milk adiponectin and its correlation with infant growth]. AB - OBJECTIVE: To investigate the variation of human breast milk adiponectin (APN) concentration during lactation, analyze the relationship of APN concentrations in human breast milk with APN in infant serum, determine the association between maternal milk APN and infant body proportionality in the first year of life, and the period of greatest human milk exposure. METHOD: Subjects included 73 mother infant pairs from Shanghai International Peace Maternity & Child Health Hospital. Maternal milk was collected at 2, 4, 13, 26 weeks and infant serum was collected at 26 weeks. The concentration of APN in maternal milk and infant serum were measured with commercially available ELISAs (R&D Systems). Weight-for-age Z scores (WAZ), length-for-age Z-scores (LAZ), weight-for-length Z-scores (WLZ) and body mass index Z-scores (zBMI) of infants up to 1 year of age were calculated using World Health Organization (WHO) standards. Then the variation of maternal milk APN was investigated, the correlation between maternal milk APN and infant serum APN was analyzed; multivariate regression analysis was used to analyze covariates correlated with infant body proportionality, meanwhile determined the association between maternal milk APN and infant anthropometrics. RESULT: The concentration of APN in maternal milk at 2, 4, 13, 26 weeks was respectively (14.62 ug/L, 5.93 - 140.40 ug/L), (7.32 ug/L, 2.04 - 29.35 ug/L), (6.84 ug/L, 2.72 - 15.65 ug/L) and (4.88 ug/L, 1.12 - 13.38 ug/L). Higher milk APN in postpartum 2 weeks was associated with lower infant WAZ at 13, 26 and 52 weeks (P < 0.05). Milk APN in postpartum 4 weeks was inversely related to infant WAZ, WLZ, zBMI at 13, 26 and 52 weeks (all P < 0.05). Meanwhile, milk APN in postpartum 13 weeks had significantly negative correlation with infant WAZ, WLZ, zBMI at 13, 26 and 52 weeks (P < 0.01). Milk APN in postpartum 26 weeks also was inversely associated with infant WAZ, WLZ and zBMI at 52 weeks (P < 0.05). After adjusting covariates like birth weight using multivariate regression analysis, milk APN level is the most important influence factor to infant WAZ, WLZ, zBMI at 13, 26 and 52 weeks. There was a positive correlation between maternal milk APN and infant serum APN at 13, 26 weeks. CONCLUSION: Milk APN concentration declines throughout lactation. Maternal milk APN may play a vital role in the growth and development of breastfed infants, particularly closely associated with infant weight. Infant serum APN clearly correlated with maternal milk APN. PMID- 21624285 TI - [A brief description of the 18th International Conference on Prevention of Child Abuse and Neglect]. PMID- 21624284 TI - [Recommendations for the diagnosis and management of food allergy in infants and young children]. PMID- 21624286 TI - [Early prediction of the injured regions in neonatal brain with hypoxic-ischemic encephalopathy by diffusion weighted imaging and measuring their apparent diffusion coefficient]. AB - OBJECTIVE: To elucidate that diffusion weighted imaging (DWI) can be used to predict the injured regions of neonatal brain with hypoxic-ischemic encephalopathy (HIE) in the early phase of injury, and to measure the apparent diffusion coefficient (ADC) values in the multiple regions of the brain. METHOD: The participants in this study were twenty-six infants with HIE from neonatology ward hospitalized between July 2006 and July 2009. Nineteen patients had severe HIE, and seven had moderate HIE. DWI and conventional magnetic resonance imaging (MRI) were performed for each case within the first 72 hrs. The ADC values of eight regions of interest (ROIs) were measured in ten cases with severe HIE (ADC values group). ROIs included posterior limb of internal capsule (PLIC), ventrolateral thalami, basal ganglia, perirolandic cortex, occipital cortex, centrum semiovale, brainstem, and frontal white matter. Twelve neonates were enrolled as the control subjects. RESULTS: During the first 72 hrs, the conventional MRI of 26 patients showed subarachnoid hemorrhage in 5, subdural hemorrhage in 2, and mild high signal intensity in the cortex of only one patient. In the 19 cases with severe HIE, abnormal signal intensities were seen in ventrolateral thalami and perirolandic cortex of 17 patients (89%), and the remaining 2 infants showed abnormal cortex and subcortical white matter. In 7 cases with moderate HIE, 4 had abnormal signal intensity in the cortex and subcortical white matter, 2 had abnormal periventricular white matter, and only one showed abnormal signal intensity in the ventrolateral thalami and perirolandic cortex. In the ADC values group, the average ADC values of posterior limb of internal capsule (PLIC), ventrolateral thalami, basal ganglia, perirolandic cortex, occipital cortex, centrum semiovale, brainstem, and frontal white matter respectively were 0.68 (0.56 - 0.88), 0.73 +/- 0.13, 0.67 +/- 0.11, 0.78 +/- 0.22, 0.90 +/- 0.16, 0.87 +/- 0.21, 0.73 +/- 0.19, 1.32 +/- 0.22 * 10( 3) mm(2)/S. In the control group, the average ADC values of posterior limb of internal capsule (PLIC), ventrolateral thalami, basal ganglia, perirolandic cortex, occipital cortex, centrum semiovale, brainstem, and frontal white matter respectively were 0.96 (0.95 - 1.02), 1.02 +/- 0.90, 1.15 +/- 0.99, 1.08 +/- 0.07, 1.09 +/- 0.08, 1.39 +/- 0.20, 0.96 +/- 0.05, 1.58 +/- 0.18* 10(-3) mm(2)/S. There was statistically significant difference in the average ADC values between each of 8 ROIs of infants with HIE and healthy neonates (P < 0.01). CONCLUSION: In the first days after birth, the major injured regions of severe HIE were ventrolateral thalami and perirolandic cortex, the minor injured regions were cortex and subcortical white matter. Multiple regions of moderate HIE were injured, including cortex with subcortical white matter, periventricular white matter, and ventrolateral thalami with perirolandic cortex. The ADC values of the regions with abnormal signal intensity decreased, also some regions with the normal signal intensity. PMID- 21624287 TI - [Analysis of clinical features, biochemical analysis and gene mutations in one Chinese pedigree with neonatal-onset ornithine transcarbamylase deficiency]. AB - OBJECTIVE: This study aimed at understanding clinical features, biochemistry and gene mutation in one Chinese pedigree which had a neonatal-onset ornithine transcarbamylase deficiency (OTCD) boy, and exploring the significance of ornithine transcarbamylase analysis in prenatal diagnosis. METHOD: The clinical and biochemical data of one case were analyzed. The amino acids in blood and organic acids in urine were analyzed by mass spectrum technology. The OTC gene mutation was detected using polymerase chain reaction (PCR) and DNA direct sequencing for the case, his parents and the fetus amniocyte and her blood after birth. RESULT: The age of onset was 3 days after birth, he began to have poor reaction, difficulty to feed, high blood ammonia, infection, slight metabolic acidosis, which were consistent with the clinical diagnosis of urea cycle disorders. The boy died at the age of 9 days. Citrulline of blood was detected twice, and were 0.86 um and 1.06 um, respectively. The orotic acid was elevated (124 um/M Creatinine), and urine lactic acid was significantly elevated. The citrulline and orotic acid in his parents and their second baby were normal in DBS and urine. One nonsense mutation in the OTC gene was found at the exon 9 (C. 958 C > T) and his mother was the heterozygote, which caused an arginine to terminate the code at position 320 of the protein (R320X). Two other mutations were also detected at intron 9 (C.1005 + 132 InsT) and intron 5 (C.542 + 134 G > G/A). But the analysis of his father's DNA, the fetus amniocyte and her blood was normal. CONCLUSION: The mutation of C. 958 C > T in OTC gene may occur during neonatal period. This mutation would result in a very severe symptom, even die suddenly several days after birth, if it was a boy. It needs more researches to discuss whether the C.1005 + 132 InsT in intron 9 and C.542 + 134 G > G/A in intron 5 were associated with the neonatal-onset OTCD. The DNA analysis of OTC gene could be utilized for the prenatal diagnosis. PMID- 21624288 TI - [Effects of hyperoxia on erythropoietin receptor expression in lung development of neonatal rats]. AB - OBJECTIVE: Oxygen toxicity is thought to be a major contributing factor in the pathogenesis of bronchopulmonary dysplasia (BPD). Animal experiments reveal that erythropoietin (EPO) may have protective effects against hyperoxic lung injury, but the mechanisms remain unknown. The aim of this study was to evaluate effects of hyperoxia on erythropoietin receptor expression in lung development of neonatal rats. METHODS: Several litters of Wistar pups were pooled together within 12 hours after birth and randomly divided into two groups (n = 24 in each): air-exposed control group and hyperoxia-exposed group. In hyperoxia exposed group, the rats were exposed to 85% oxygen. Pups (n = 8) from each group were sacrificed on postnatal days 3, 7, and 14. The pulmonary histological and morphometric changes were observed after hematoxylin-eosin (HE) staining under light microscope. Radical alveolar counts (RAC) were compared between the two groups to evaluate the differences of alveolarization. Expressions of platelet endothelial cell adhesion molecule-1 (PECAM-1) and erythropoietin receptor (EPOR) in lung tissue were measured by immunohistochemistry. Expressions of EPOR mRNA and EPOR protein were measured by RT-PCR and Western blotting. RESULTS: In hyperoxia-exposed group, there were a few inflammatory cells infiltration in interstitium on day 3 and inflammatory response worsened on day 7. Alveolar and capillary hypoplasia and interstitial fibrosis were evident on day 14. RAC increased in air-exposed control group along with the age in days. RAC decreased from day 7 in hyperoxia-exposed group compared with air-exposed control group [(6.85 +/- 1.04) vs. (7.33 +/- 1.0), P < 0.01], which was more evident on day 14 [(6.20 +/- 1.58) vs. (9.07 +/- 0.69), P < 0.001]. Expression of PECAM-1 protein increased in air-exposed control group along with the age in days. But in hyperoxia-exposed group, it decreased on day 7 and 14 [(15.14 +/- 1.51) vs. (31.47 +/- 2.43), (11.04 +/- 1.76) vs. (41.41 +/- 3.83), P < 0.001] compared with air-exposed control group. Expression of EPOR on day 3 in air-exposed control group was the strongest and weakened gradually with the increase of postnatal days. Expression of EPOR in hyperoxia-exposed group decreased on day 3 and became more evident on day 7 and day 14 compared with air-exposed control group [(1.62 +/- 0.04) vs. (1.82 +/- 0.06), P < 0.05; (0.48 +/- 0.01) vs. (1.10 +/- 0.07), (0.39 +/- 0.04) vs. (0.87 +/- 0.03), P < 0.001]. Expression of EPOR mRNA on day 3 in air-exposed control group was the strongest and was decreased significantly in hyperoxia-exposed group compared with air-exposed control group at all time points [(0.87 +/- 0.07) vs. (1.1 +/- 0.17), (0.18 +/- 0.07) vs. (0.36 +/- 0.08), P < 0.01;(0.14 +/- 0.05) vs. (0.36 +/- 0.09), P < 0.001]. CONCLUSIONS: EPOR may participate in the modulation of normal lung development. Depressed expression of EPOR and PECAM-1 may be involved in the pathogenesis of alveolar and capillary hypoplasia induced by hyperoxia. PMID- 21624289 TI - [Value of serum cytosolic beta-glucosidase in diagnosis of neonatal necrotizing enterocolitis]. AB - OBJECTIVE: This study was conducted to compare serum cytosolic beta-glucosidase (CBG) levels of age-matched control patients with those of infants with necrotizing enterocolitis (NEC) thereby to determine the eventual association between serum CBG levels with extensive disease in infants with NEC. METHOD: A total of 96 premature infants were divided into the early NEC group (n = 25), confirmed NEC group (n = 23) and the control group (n = 48). Serum CBG concentration, C-reactive protein (CRP) and peripheral blood white blood cells (WBC) were measured at the onset of the disease in patients in early NEC or confirmed NEC groups and at weeks 2-3 in control infants. Data were analyzed using descriptive statistics, non-parametric tests, Student's t-test, linear correlation, Spearman correlation analysis, receiver operating characteristic (ROC) curve were used for statistical analysis. RESULTS: The median birth weights (mean +/- SE) in the three groups were not statistically significant (P > 0.05). Serum CBG concentration in the 3 groups were (112.369 +/- 108.539) nmol/L, (693.013 +/- 211.614) nmol/L and (36.478 +/- 28.31) nmol/L, respectively. The infants in the confirmed NEC group had highest CBG levels, compared with the other 2 groups (P < 0.05). When the levels of CBG >= 65 ng/ml, CRP >= 2 mg/L and WBC < 5 * 10(9)/L within 3 days after birth or > 20 * 10(9)/L 3 days after birth were considered as positive parameters, the sensitivity of CBG and CRP was higher than that of WBC (P < 0.05). Among these indices, CBG had the highest specificity (87.4%), positive predictive (95.6%) and Youden's index (81.3%). CBG is correlated with CRP (the Spearman correlation coefficient was 0.379, P < 0.01). CONCLUSION: Serum CBG concentration increases early in NEC. Serum CBG level was associated with extensive disease in infants with NEC. Therefore CBG can be used as a marker in the early diagnosis of NEC. PMID- 21624290 TI - [Childhood-onset myasthenia gravis: the analysis of influencing factors of therapeutic effect and prognosis]. AB - OBJECTIVE: Though myasthenia gravis (MG) is a typical autoimmune disorder, there was some controversy on the treatment of the childhood-onset MG. By observing the efficacy of different therapies, the authors analyzed the affecting factors of prognosis in childhood-onset MG. METHOD: The retrospective data of 155 patients with childhood-onset MG (age of MG onset was less than 15 years) were collected from Department of Neurology, Beijing Tongren Hospital (January 2000 - February 2010). The patients were non-randomly divided according to their treatment into 3 groups (glucocorticoid, thymectomy and glucocorticoid combined with thymectomy groups). Postintervention status meeting the criteria of Myasthenia Gravis Foundation of America (MGFA) "complete stable remission, CSR", "pharmacologic remission, PR", "minimal manifestations, MM", or "Improved, I" was regarded as desirable response, which was used as primary indicator of observation. The authors assessed the efficacy of three therapies and analyzed the influencing factors of prognosis by using Chi-square test and Logistic regression. RESULT: At 3 months of treatment, glucocorticoid group showed the highest effective rate. At the end of 1 year or 2 years of treatment, glucocorticoid combined with thymectomy group showed the highest effective rate respectively. The generalization rate of MG at 2 years, 10 years and 20 years in childhood-onset ocular MG patients were 4.3%, 10.7%, and 41.5%, respectively. Of patients with generalization of MG, 48.1% occurred within 2 years, 92.6% within 20 years. Univariate analysis showed that in childhood-onset ocular MG patients, variables such as age at onset (> 10 years), LG-MG and with chronic fatigue were significantly associated with general MG conversion. Whereas multivariate analysis showed that patients with age at onset (> 10 years) and chronic muscle fatigue were apt to convert to generalized MG. CONCLUSION: Glucocorticoid appeared to have an effect that leads to early remission of symptoms in childhood onset MG patients and glucocorticoid combined with thymectomy appeared to have better long-term effect. For those childhood-onset ocular MG patients with longer course of disease, older age of onset, chronic fatigue, or LG-MG, physicians should try to prevent the generalization of MG by immunosuppressive therapies. PMID- 21624291 TI - [Effects of carvedilol on cardiomyocyte apoptosis in autoimmune myocarditis in mice]. AB - OBJECTIVE: To observe the effects of carvedilol on the expression of Bcl-2, Bax and Fas in autoimmune myocarditis (AM). METHODS: A total of 60 inbred male BALB/C mice 4 - 5 weeks of age were divided at random into 3 groups as follows: AM group (n = 20), carvedilol group (n = 20) and control group (n = 20). The mice were sacrificed after gathering blood specimens by taking out the eyeballs and hearts tissue. The histological and ultrastructural changes were observed under light microscope and electron microscope. The concentrations of cardiac troponin I (cTn I) were detected by chemiluminescence immunoassay (CLIA). Immunohistochemistry (IHC) was performed to analyze the contents of Bcl-2, Bax and Fas, TUNEL to detect the apoptotic index in myocardial cells. RESULTS: There were large number of lymphocyte and monocyte infiltrates under light microscope and karyopyknosis and chromatin gathered along the nuclear membrane under electron microscope in AM group. There were no inflammations and chromatin gathering in group C. Compared with control group, the Bcl-2, Bax and Fas protein expression significantly elevated in AM group (23.48 +/- 2.24 vs. 6.64 +/- 1.60, 26.15 +/- 2.02 vs. 5.09 +/- 0.85, 21.22 +/- 3.62 vs. 5.86 +/- 1.37, P < 0.01). The histopathologic scores (2.60 +/- 0.31 vs. 2.02 +/- 0.26, P < 0.05) and karyopyknosis of carvedilol group decreased as compared with AM group. The Bcl-2, Bax and Fas protein expression (17.13 +/- 1.94 vs. 23.48 +/- 2.24, 17.66 +/- 2.62 vs. 26.15 +/- 2.02, 16.79 +/- 2.83 vs. 21.22 +/- 3.62, P < 0.05), AI [(16.61 +/- 4.67)% vs. (24.51 +/- 4.70)%, P < 0.05] and contents of cTnI [(1.878 +/- 0.48) ng/ml vs. (1.102 +/- 0.23) ng/ml, P < 0.05] also decreased in carvedilol group compared with AM group. CONCLUSION: Carvedilol could protect against AM by alleviating cardiomyocyte apoptosis. PMID- 21624292 TI - [A case of coloboma of optic nerve in newborn]. PMID- 21624293 TI - [Non-drug treatment for hypertrophic obstructive cardiomyopathy in children]. AB - OBJECTIVE: To retrospectively summarize the effect of non-medical therapies for pediatric patients with hypertrophic obstructive cardiomyopathy (HOCM). METHODS: From Nov. 2008 to Jun. 2010, 4 children with drug-refractory HOCM were admitted to our hospital. Their ages were 14, 7, 9 and 6 years old, respectively. Their body weights were 38, 17, 21.5 and 17 kg, respectively. Before operation, the pressure gradients over left ventricular outflow tract (LVOTG) were 60, 147, 58 and 114 mm Hg (1 mm Hg = 0.133 kPa), respectively. And mitral regurgitation (MR) areas were 2.2, 7.3 cm(2) and 2.9 cm(2), respectively, except that it was trivial in one case. Percutaneous transluminal septal myocardial ablation (PTSMA) was performed in case 1 and 2. Septal myectomy (SM) was performed in case 3 and 4. Follow-up was first performed right after operation or before discharge, then 1 month, 3 months, 6 months, and 12 months after operation, and then once a year. The follow-up period was 1 - 18 (9.3 +/- 8.1) months. RESULTS: All patients experienced relieved symptoms. Three of them had their NYHA functional class improved except case 2. Echocardiography revealed that LVOTGs right after operations were 38, 79, 20 and 0 mm Hg, respectively, suggesting significant improvement of left ventricular outflow tract obstruction (LVOTO) in all patients. During follow-up, case 2 suffered from recurrence of LVOTO, while the other 3 cases showed sustained relief. In the last follow-up, the LVOTGs of the four patients were 19, 168, 16 and 0 mm Hg, respectively. Echocardiography also revealed that MRs of all patients were significantly reduced, even in case 2 whose LVOTG rebounded, with no recurrence during follow-up. Severe complications were absent, such as ventricular septum perforation, cardiac tamponade, ventricular tachycardia or ventricular fibrillation. No one suffered from complete heart block. Transient complete right bundle branch block (CRBBB) was observed in case 1 after PTSMA and converted to intraventricular block after 1 month. Complete left bundle branch block (CLBBB) was present in both case 3 and 4, who received SM. In case 4, it converted to intraventricular block after 1 month while in case 3 CLBBB persisted. CONCLUSIONS: The initial experience showed that PTSMA and SM were safe and effective for drug-refractory symptomatic HOCM children, with satisfactory short-term results. Further studies are needed to evaluate the long-term results and complications. PMID- 21624294 TI - [Dyspnea complicated with pleural effusion in a newborn infant]. PMID- 21624295 TI - [The issue of methylprednisolone sodium succinate dosage for use]. PMID- 21624296 TI - [A case of neonatal protein C deficiency]. PMID- 21624297 TI - [Analysis of two cases with childhood intestinal Behcet's disease]. PMID- 21624298 TI - [Advances in arrhythmogenic right ventricular cardiomyopathy]. PMID- 21624299 TI - [Endothelial progenitor cells and the neovascularization in patients with coronary aneurysms due to Kawasaki disease]. PMID- 21624300 TI - [Interpretation of task force recommendations for prevention and treatment of acquired long-QT syndrome]. PMID- 21624301 TI - [Clinical characteristics of 52 patients with torsade de points associated with acquired QT prolongation]. AB - OBJECTIVE: Risk factors, ECG characteristics and treatment options of patients with Torsade de Points associated with acquired QT prolongation are summarized in this study. METHOD: Using "torsade de points" and "QT prolongation" as the keywords to search the inpatients database from 1990 - 2010 of Fuwai hospital, 52 eligible patients were included in this analysis. RESULTS: Structural heart diseases were found in 67.3% and electrolyte disorders in 59.6% patients, 36.5% patients received diuretic therapy and 28.8% received antiarrhythmic drugs which might induce prolonged QT interval. The mean QTc was (571 +/- 93) ms and (456 +/- 50) ms before and after treatment. All patients received potassium and magnesium supplement. Isoproterenol was used in 32.7% patients. 13.5% patients received temporary pacing therapy. CONCLUSIONS: Torsade de points and acquired QT interval prolongation was often associated with electrolyte disorders and drugs causing QT prolongation. ECG and QTc should be intensively monitored for high risk patients. Early awareness of the warning signs might contribute to early recognition and proper treatment of patients with Torsade de Points associated with acquired QT prolongation. PMID- 21624302 TI - [Clinical characteristics and outcome of 32 patients with long-QT syndrome accompanied with torsade de pointes]. AB - OBJECTIVE: To summarize the clinical characteristics and outcome of patients with long-QT syndrome (LQTs) accompanied with torsade de pointes. METHODS: Thirty-two eligible patients were included in this study. Clinical and electrocardiographic data were analyzed and telephone or out-patient follow-up were made in all patients. RESULTS: There were 15 patients with inherited LQTs (h-LQTs) and 17 patients with acquired LQTs (a-LQTs). There are more women (n = 24) than men (n = 8). beta blockers, potassium and magnesium supplement were the basic therapy for h-LQTs patients, bivent pacemaker was implanted in 2 patients and implantable cardioverter defibrillator was implanted in 5 patients. Ventricular tachyarrhythmias and syncope occurred in 4 patients during (39.4 +/- 25.1) months follow-up. In 17 a-LQTs patients, one patient with dilated cardiomyopathy died suddenly and another patient with implanted cardioverter defibrillator experienced one ventricular tachycardia during (30.9 +/- 13.3) months follow-up. CONCLUSIONS: The prognosis in h-LQTs and a-LQTs patients with structure heart disease is poor. ICD or CRT-D therapy is suggestive for a-LQTs patients with structure heart disease. PMID- 21624303 TI - [Effect of antiarrhythmic peptide on ventricular arrhythmia induced by lysophosphatidic acid]. AB - OBJECTIVE: To investigate the effect and potential mechanism of lysophosphatidic acid (LPA) and antiarrhythmic peptide (AAP10) on rabbit ventricular arrhythmia. METHODS: Twenty-four rabbits were randomly divided into three groups (n = 8 each): control group, LPA group and AAP10 + LPA group. Using arterially perfused rabbit ventricular wedge preparations, transmural ECG and action potentials from both endocardium and epicardium were simultaneously recorded in the whole process of all experiments with two separate floating microeletrodes. The incidence of ventricular arrhythmia post S1S2 stimulation was recorded. Protein levels of nonphosphorylated Cx43 and total Cx43 were evaluated by Western blot. The distribution of nonphosphorylated Cx43 was observed by confocal immunofluorescence microscopy. RESULTS: Compared with the control group, the QT interval, endocardial action potential duration, transmural repolarization dispersion (TDR) and incidence of ventricular arrhythmia were significantly increased and nonphosphorylated Cx43 expression was significantly upregulated in the LPA group. Compared with the LPA group, cotreatment with AAP10 can reduce the QT interval, endocardial action potential duration, TDR and incidence of ventricular arrhythmia (25.0% vs 62.5%, P < 0.01) and downregulate nonphosphorylated Cx43. CONCLUSIONS: LPA could promote the arrhythmia possibly by upregulating nonphosphorylated Cx43 and subsequent gap junction transmission inhibition. Gap junction enhancer AAP10 could attenuate the pro-arrhythmic effect of LPA probably by downregulating myocardial nonphosphorylated Cx43 expression. PMID- 21624304 TI - [Efficacy and safety of intravenous recombinant human brain natriuretic peptide in patients with decompensated acute heart failure: a multicenter, randomized, open label, controlled study]. AB - OBJECTIVE: To explore the safety of intravenous recombinant human brain natriuretic peptide (rhBNP) in treating acute decompensated heart failure and acute exacerbation of chronic heart failure, and to compare the differences in efficacy with different dosage and administration time. METHODS: A total of 2160 patients characterized of acute decompensated heart failure and acute exacerbation of chronic heart failure were enrolled in this multicenter, randomized, open, dose-control study. The patients were randomly allocated to four groups with different doses and administration time on top of standard therapy. RESULTS: In the safety respect, the rate of hypotension is 1.44% at 5 - 7 days after treatment, the serum creatinine level was reduced compared to baseline (P values were 0.0437, 0.0087 and 0.0116) except in the group of 0.015 ug at 24 h (P = 0.7054). The rate of 30-day readmission is 5.65%, mortality rate is 9.44%. In terms of efficacy, dyspnea was significantly improved at 30 min after administration, and at 24 h after administration (all P < 0.01). Urine output and LEVF were also significantly increased by 76.59% (P < 0.01) and 12.08% respectively (all P < 0.01) compared to baseline. Plasma NT-proBNP decreased by 40.29% at 5 - 7 days after administration (P < 0.01). CONCLUSION: The clinical application of intravenous rhBNP is safe and effective for treatment of acute decompensated heart failure and acute exacerbation of chronic heart failure in this large patient cohort. PMID- 21624305 TI - [Modeling of systolic blood pressure reaction to antihypertensive agents in people with hypertension]. AB - OBJECTIVE: We used the individual patient data from clinical trials, pooled in the INDANA data set, to explore whether blood pressure reduction was related to the baseline individual characteristics, and quantify the potential associations. METHODS: We used the data from 31 140 patients with essential hypertension recruited in four randomized placebo-controlled clinical trials, MRC35-64, MRC65 74, STEP and SYST-EU. Thiazide diuretics, beta-blocker, and calcium channel blocker, three of six major BP lowering drugs were analyzed. Patients were all with the same first dosage of the drug in each trial. Age, body weight, height, level of total cholesterin (TC), systolic blood pressure (SBP) and diastolic blood pressure (DBP) when initialed and at first visit of follow-up, pharmacological treatment, gender, status of smoking, history of myocardium infarction were factors taken into model. Data were managed by software SAS((r)). Statistical analyses were performed with SAS((r)) and R. Model was developed to evaluate the relationship between decrease of SBP and characteristics of patients. RESULTS: Initial SBP is the only modifier of treatment effect on SBP response in the 3 BP lowering drug classes (beta = 0.09, 0.37 and 0.18, respectively). Age and initial DBP were factors significantly correlated with SBP fall for diuretic (beta = 0.17 and 0.14), and age was one of factors significantly correlated with SBP fall for beta-blocker (beta = -0.17). Smokers would receive less SBP fall compare to non-smokers in beta-blocker active treated group (beta = -2.07). There is converse effect of age between the diuretic and beta-blocker; older people seem sensitive to diuretic, while young people are sensitive to beta-blocker. As to calcium channel antagonist class, body weight is another modifier (beta = 0.06) (All P value are 0.000 except 0.050 for body weight in calcium channel antagonist class). CONCLUSION: We identified 5 significant modifiers (baseline SBP and DBP, age, smoking status and body weight) for SBP response to treatment effect, while gender, TC and history of myocardial infarction are not modifiers for SBP response to treatment effect. PMID- 21624306 TI - [Risk factors affecting in-hospital mortality of the arterial switch operation for transposition of the great arteries]. AB - OBJECTIVE: To analyze the in-hospital mortality and factors affecting in-hospital mortality for patients with transposition of the great arteries (TGA) undergoing arterial switch operation (ASO). METHODS: Between January 2004 and December 2007, ASO was performed in 169 patients [129 male, 40 female; mean age (11.71 +/- 26.3) months] with TGA. The patients were divided in intact ventricular septum group (n = 56): TGA with intact ventricular septum and ventricular septal defect group (n = 113): TGA with ventricular septal defect. Multiple logistic regression analysis was performed to identify the risk factors of in-hospital mortality. RESULTS: The overall in-hospital mortality was 11.24% (19/169). The yearly in-hospital mortality was similar between intact ventricular septum group and ventricular septal defect group. With the improvement of perioperative treatment, the in hospital mortality decreased from 16.67% in 2004 to 3.92% in 2007. The multivariate analysis revealed that body weight <= 3 kg (OR: 4.571, P = 0.0409), complicating ventricular septal defect (OR: 4.444, P = 0.0406), complex TGA (OR: 4.321, P = 0.0140), coronary anomalies (OR: 4.867, P = 0.0104) and non-type A coronary arteries (OR: 3.045, P = 0.0243) were independent predictors for poor early postoperative survival. CONCLUSION: Body weight <= 3 kg, complicating ventricular septal defect, complex TGA, coronary anomalies are independent predictors for increased in-hospital mortality in patients with transposition of TGA and undergoing arterial switch operation. PMID- 21624307 TI - [Association between serum amyloid protein A1 polymorphisms and carotid intima media thickness in Han Chinese]. AB - OBJECTIVE: To explore the association between genetic polymorphism of serum amyloid protein A1 (SAA1) with carotid intima media thickness in a healthy Han Chinese population of Xinjiang. METHODS: A total of 449 healthy Han Chinese participating the cardiovascular risk survey between June 2007 and September 2009 were included, the genotypes of the SAA1 were detected by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). The mean IMT of the right and left common carotid arteries were measured by B-mode ultrasonography. RESULTS: (1) There was strong linkage disequilibrium between rs12218 and rs2229338 (D' = 0.89). (2) The carotid common IMT (CC-IMT) and the carotid bulb IMT (CB-IMT) were similar between the AA genotype (wild genotype) and the GGFAG genotype (mutational genotype) in rs2229338 of SAA1 gene. (3) CC IMT [(0.081 +/- 0.071) cm vs (0.068 +/- 0.019) cm, P = 0.01] was significantly thicker in CC + CT genotype (mutational genotype) group than in TT genotype (wild genotype) of rs12218 group and the difference remains significant after adjustment for age, gender, blood pressure, waist circumference, creatinine and high density lipoprotein cholesterol. CB-IMT [(0.085 +/- 0.038) cm vs. (0.081 +/- 0.052) cm, P = 0.36] was similar between CC + CT genotype and TT genotype of rs12218 groups. CONCLUSION: Our results suggested that the genetic polymorphism of SAA1 might be linked with IMT and rs12218 mutation could serve as a promoting factor for IMT in Han Chinese people. PMID- 21624308 TI - [Implication of N-terminal pro-B-type natriuretic peptide and Tei index of left ventricle changes in children with ventricular septal defect treated by transcatheter interventional therapy]. AB - OBJECTIVE: To explore the implication of the dynamic changes of plasma N-terminal pro-B-type natriuretic peptide (NT-proBNP) level and Tei index of left ventricle (LV) in children with ventricular septal defect (VSD) treated by transcatheter closure. METHODS: Sixty children with VSD treated by transcatheter closure with VSD occluder (Group VSD) and 30 healthy children (Group C) were included in this study. The plasma concentration of NT-proBNP, Tei index of LV and left ventricle ejection fraction (LVEF) were measured in Group C and at before, 5th minute, 4th hour, 1st month, 3rd month and 6th month after VSD closure in Group VSD. RESULTS: (1) The concentration of plasma NT-proBNP was significantly increased in children with VSD before transcatheter closure compared with Group C [(229.45 +/- 57.75) ng/L vs. (99.21 +/- 46.86) ng/L, P < 0.01], significantly increased at 5th minute and 24th hour after transcatheter closure [(356.27 +/- 96.78) ng/L and (356.38 +/ 91.95) ng/L vs. (229.45 +/- 57.75) ng/L, all P < 0.01], and significantly decreased at 1st month, 3rd months and 6th months after transcatheter closure [(131.33 +/- 34.79) ng/L, (96.56 +/- 31.55) ng/L and (93.39 +/- 29.46) ng/L vs. (229.45 +/- 57.75) ng/L, P < 0.05 or P < 0.01]. (2) The Tei indexes of LV in Group VSD before transcatheter closure were significantly higher than in Group C (0.45 +/- 0.05 vs. 0.33 +/- 0.08, P < 0.01) and Tei index was significantly increased at 24th hour, 1st month after transcatheter closure (P < 0.01) while significantly decreased at 3rd and 6th month compared with those before transcatheter closure (0.34 +/- 0.07 and 0.34 +/- 0.06 vs. 0.45 +/- 0.05, all P < 0.01). (3) There is a positive correlation between the changes of the plasma concentration of NT-proBNP and the change of Tei index of LV before and after transcatheter closure (r = 0.653, P < 0.05). CONCLUSION: Tei index of LV and NT proBNP can monitor cardiac function changes in children with VSD before and after transcatheter closure. PMID- 21624309 TI - [Clinical application and value of 3 Tesla contrast enhanced whole-heart coronary magnetic resonance angiography]. AB - OBJECTIVE: To investigate the clinical application, feasibility and value of 3 T whole-heart contrast enhanced free-breathing navigator-gated three-dimensional coronary magnetic resonance angiography (CE-CMRA). METHODS: 3 T CE-CMRA was used to examine patients with suspected coronary heart disease (CAD). Gd-BOPTA (0.2 mmol/kg) was injected intravenously with slow infusion rate (0.3 ml/s) to perform enhancement. Data were post-processed to obtain principal branches of coronary artery and picture quality was evaluated. According to results of selective coronary arteriography (SCAG), the diagnostic accuracy of CE-CMRA for diagnosing CAD was judged by means of detecting significant stenosis (> 50%) of the principal branches based on the 9 segments of coronary artery. RESULTS: Twenty three out of 26 patients successfully completed the examination. The mean scanning time was (10.4 +/- 2.1) minutes, 178 out of 202 (88.1%) SCAG demonstrated segments could be evaluated by CE-CMRA. The imaging quality was superior in proximal and middle segments of coronary artery principal branches than in distal segments. Based on patient-level, there were 9 positive cases and 14 negative cases examined by CE-CMRA compared with 11 positive cases and 12 negative cases examined by SCAG, respectively. The whole diagnose accordance rate of CE-CMRA was 91.3% (21/23) compared with SCAG. The sensitivity, specificity and negative predictive values were 81.8% (9/11), 88.5% (169/191) and 98.8% (9/31) respectively. CONCLUSIONS: 3 T CE-CMRA is a feasible non-invasive imaging modality for diagnosing CAD, especially to detect significant stenosis in proximal and middle segments of coronary artery principal branches. However, the detecting efficacy is limited in assessing stenosis of distal segment and small branches of coronary artery. PMID- 21624310 TI - [Effects of sarcoplasmic reticulum Ca(2+)-ATPase gene transfer in a minipig model of chronic ischemic heart failure]. AB - OBJECTIVE: Chronic myocardial ischemia (CMI) has become the most important cause of heart failure (HF) all over the world. The aim of the current study was to investigate the effects of Sarco-endoplasmic reticulum calcium ATPase 2a (SERCA2a) gene transfer on cardiac function and endoplasmic reticulum stress (ERS) associated myocardial apoptosis in a minipig HF animal model induced by CMI. METHODS: HF was induced in minipigs by implantation of ameroid constrictor in the initial segment of left anterior descending (LAD) branch of coronary artery. After confirmation of myocardial perfusion defects and cardiac function impairment by myocardial perfusion imaging and echocardiography, animals were divided into 4 groups (n = 4 each): HF group, HF + enhanced green fluorescent protein (EGFP) group, HF + SERCA2a group, and shamed animals as control group. A total amount of 1 * 10(12) v.g. of rAAV1-EGFP or rAAV1-SERCA2a were injected intramyocardially to each animal of HF + EGFP and HF + SERCA2a groups. Sixty days after gene transfer, protein level and activity of SERCA2a were examined, cardiac functions and changes of serum inflammatory and neuro-hormonal factors were determined. Apoptotic index of the ischemic myocardium, protein levels of ER stress marker glucose regulated protein 78 (GRP 78) and ER stress specific apoptotic marker caspase-12 were also assayed. RESULTS: At the study end, echocardiographic and hemo dynamic measurements indicated a significant improvement of both cardiac systolic and diastolic function in HF + SERCA2a group compared with HF/HF + EGFP groups [LVEF (60.2 +/- 8.6)% vs (44.2 +/- 7.1)% and (46.8 +/- 6.7)%, Ev/Av 1.28 +/- 0.24 vs 0.77 +/- 0.17 and 0.80 +/- 0.21, +dp/dt(max) (2713.9 +/- 434.0) mm Hg/s (1 mm Hg = 0.133 kPa) vs (1892.3 +/- 434.2) mm Hg/s and (1931.2 +/- 397.4) mm Hg/s, -dp/dt(max) (1422.1 +/- 334.4) mm Hg/s vs (848.3 +/- 308.3) mm Hg/s and (849.5 +/- 278.3) mm Hg/s, P < 0.05], along with increase in both SERCA2a protein level (1.13 +/- 0.26 vs 0.73 +/- 0.17 and 0.64 +/- 0.18, P < 0.05) and activity [(16.2 +/- 5.5) IU/ml vs (7.9 +/- 3.1) IU/ml and (7.5 +/- 2.8) IU/ml, P < 0.05] compared with HF/HF + EGFP groups. Serum concentrations of inflammatory factor tumor necrotic factor alpha [(382.3 +/- 114.4) ng/L vs (732.3 +/- 201.4) ng/L and (689.8 +/- 192.5) ng/L, P < 0.05], neural-hormonal factors brain natriuretic peptide [(142.6 +/- 45.3) ng/L vs (422.3 +/- 113.6) ng/L and (393.7 +/- 103.3) ng/L, P < 0.01], endothelin-1 [(111.4 +/- 37.5) ng/L vs (193.5 +/- 54.3) ng/L and (201.0 +/- 72.1) ng/L, P < 0.05] and angiotensin II [(189.7 +/- 65.2) ug/L vs (538.3 +/- 135.2) ug/L and (525.5 +/- 144.1) ug/L, P < 0.01] were also significantly decreased in HF + SERCA2a group compared with HF/HF + EGFP groups. The apoptotic index [(12.71 +/- 4.11)% vs (23.22 +/- 7.23)% and (24.31 +/- 6.38)%, P < 0.05], protein levels of GRP 78 (1.27 +/- 0.33 vs 3.23 +/- 1.14 and 4.18 +/- 1.13, P < 0.05) and protein level ratios of cleaved caspase-12 to total caspase-12 [(4.62 +/- 1.93)% vs (9.71 +/- 2.70)% and (10.14 +/- 2.81)%, P < 0.05] were also significantly reduced in the ischemic myocardium of HF + SERCA2a group compared with the HF/HF + EGFP groups. CONCLUSION: Overexpression of SERCA2a significantly improved cardiac systolic and diastolic function in this HF model partly through attenuation of ER stress related myocardial apoptosis, suggesting its therapeutic potential for CMI related heart failure. PMID- 21624311 TI - [Predictors of vulnerable atherosclerotic plaques induced by cholesterol and balloon injury in rabbits]. AB - OBJECTIVE: To detect the optimal predictors of vulnerable atherosclerotic plaques. METHODS: Forty New Zealand white rabbits underwent balloon-induced abdominal aortic wall injury and were fed a high cholesterol and saturated fat diet containing 1% cholesterol for 8 weeks. Rabbits were then randomly divided into two groups: group A (n = 20, the aortic segments rich in plaques were incubated transluminally with recombinant adenovirus carrying p53) and group B [n = 20, incubated transluminally with beta galactosidase (LacZ) genes]. Two weeks later, rabbits underwent pharmacological triggering with injection of Chinese Russell's viper venom (CRVV) and histamine. Before pharmacologically triggering, concentrations of hs-CRP, sVCAM-1 and sICAM-1 were measured by means of Enzyme linked-immunosorbent assay (ELISA). Fibrinogen was analyzed by nephelometer. Ultrasound imaging, accuracy densitometry (AD) examination and intravascular ultrasound (IVUS) were performed to analyze the in vivo features of vulnerable plaques. Logistic regression was used to detect the predictors for vulnerable plaques. RESULTS: The ratio of plaque rupture after pharmacological triggering was significantly higher in group A (89.5%, 17/19) than in group B (22.2%, 4/18). Serum hs-CRP level was significantly higher in plaque rupture group than in non rupture group before pharmacological triggering (P < 0.05). In the meantime, parameters derived from ultrasound imaging [intima-media thickness (IMT) and peak velocity (VP), values of accuracy densitometry], measurements of IVUS [external elastic membrance area (EEMA), plaque area (PA), plaque burden (PB), eccentric index (EI) and remodeling index (RI)] were significantly larger in plaque rupture group than in non-rupture group. Logistic regression showed that EI (OR = 26.917), PA (OR = 19.301), sVCAM-1 (OR = 1.339) and AII-c% (OR = 0.458) were independent predictors for plaque rupture (all P < 0.05). CONCLUSION: The major predictors of vulnerable plaques were eccentric index (EI) and plaque area (PA), sVCAM-1 and AII-c% in this model. PMID- 21624312 TI - [Mechanism related to docosahexaenoic acid induced large conductance calcium activated potassium channel currents increase in coronary smooth muscle cells]. AB - OBJECTIVE: To investigate the mechanism of enhanced large conductance calcium activated potassium channel currents (BK) in coronary smooth muscle cells (SMCs) by docosahexaenoic acid (DHA). METHODS: Coronary SMCs were isolated by enzyme digestion. Potassium channels in coronary SMCs were identified by applications of different potassium blockers. Effects of DHA and its metabolite 16, 17 epoxydocosapentaenoic acid (16, 17-EDP) on BK channels in the absence and presence of cytochrome P450 epoxygenase inhibitor SKF525A were studied by patch clamp in whole-cell configuration. RESULTS: BK channels were widely distributed in SMCs, and BK currents in normal SMCs accounted for (64.2 +/- 2.7)% of total potassium currents (n = 20). DHA could activate BK channels, and its 50% effective concentration (EC(50)) was (0.23 +/- 0.03) umol/L, however, the effect of DHA on BK channels was abolished after SMCs were incubated with cytochrome P450 epoxygenase inhibitor SKF525A. 16, 17-EDP, a metabolite of DHA, could reproduce the effects of DHA on BK channels, and its EC(50) was (19.7 +/- 2.8) nmol/L. CONCLUSION: DHA and metabolites can activate BK channels and dilate coronary arteries through activating cytochrome P450 epoxygenase pathway. PMID- 21624313 TI - [MicroRNA-144 over-expression induced myocytes apoptosis]. AB - OBJECTIVE: To investigate the effects of microRNA-144 (miR-144) expression on H9C2 (2-1) myocytes. METHODS: MiR-144 was up-regulated in primary cultured H9C2 (2-1) myocytes through transfection. Cells transfected with Lipofectamine(TM) 2000 and its mixture with miRNA synthesized randomly as blank control and negative control respectively. The up-regulation of miR-144 was confirmed by real time PCR. Cell apoptosis was evaluated by means of CCK-8, Caspase-3 and flow cytometry. RESULTS: Real-time PCR results showed that the miR-144 expression was obviously increased in miR-144 up-regulation group (2178.84 +/- 838.52) compared with negative (2.06 +/- 0.73) and blank (1.00 +/- 0.00) control group (all P < 0.01). The proliferation was lower, the activity of Caspase-3 was elevated and the apoptosis rates were significantly increased in miR-144 up-regulation group compared with negative and blank control group, while no significant difference was found between the latter 2 groups. CONCLUSION: MiR-144 mimics may selectively up-regulate the expression of miR-144 in myocardial cells and consequently promote apoptosis and inhibit proliferation in myocardial cells. PMID- 21624314 TI - [Metabolic syndrome is an independent risk factor for cardiovascular disease events in patients with ischemic stroke]. AB - OBJECTIVE: To explore the association between metabolic syndrome (MS) and risk of cardiovascular disease events (CVD) in patients with ischemic stroke. METHOD: A total of 1087 patients with ischemic stroke were enrolled from 5 community-based medical centres and underwent baseline evaluation on risk factors of stroke during the period of Jan. 2003 to Dec. 2006. After baseline survey, all patients were followed up until Dec 31, 2008 and new CVD events were recorded. MS was defined using CDS criteria. Proportional hazard models were used to assess the HRs and 95% CI of CVD events associated with MS and other components. RESULTS: The prevalence of MS was 40.4% at baseline. During an average follow-up of 3.5 years, 178 patients developed new CVD events. After adjusted for age, gender, smoking, drinking, marriage status, education level, hospitalization, recurrence of stroke, stroke duration, depression, cognition impairment and ADL, MS remains the independent predictor for the risk of CVD events. Compared with patients with non-MS, the risk of CVD events increased by 44% (HR: 1.44, 95%CI: 1.06 - 1.95). The risk of CVD also increased with the number of MS components. Compared with patients with 1 or less than 1 components of MS, the risk of CVD events increased by 30% (HR: 1.30, 95% CI: 0.83 - 2.04) in those with 2 components and by 69% (HR: 1.69, 95%CI: 1.11 - 2.56) in those with 3 or more components of MS. Hypertension and hyperglycemia and impaired fasting glucose also served as independent risk factors for CVD event (all P < 0.001). CONCLUSIONS: MS was independently associated with increased risk of CVD events in patients with ischemic stroke. There was a dose-response relationship between the numbers of MS components and the risk of CVD event. PMID- 21624315 TI - [Pacemaker/ICD lead extraction with the excimer laser sheath: a single center experience]. AB - OBJECTIVE: The aim of this study is to investigate the safety and effectiveness of Excimer laser-assisted lead extraction. METHODS: Consecutive lead extraction in 16 patients using the excimer laser sheath from Veterans Administration Medical Center, University of Minnesota between March 2008 and December 2009 were analyzed retrospectively. The clinical characteristics including basic disease, cardiac function, indication and reason for lead removal, lead characteristic, and complication were summarized. RESULTS: Thirty one leads (laser sheaths were used in 25 leads) in 16 patients [mean age (67.4 +/- 9.1) years, range 54 - 82 years] were extracted. All leads were extracted successfully without complication. Most patients had severe heart diseases with left ventricular ejection fraction of 20% - 60% (mean 36.4% +/- 13.4%). Mean implant time was (73.9 +/- 62.9) months (range 3 - 251 months). Nine patients had Class I indications for lead extraction, 7 patients for Class II indications. Atrial lead fractured in one patient during lead extraction, the remaining part of the lead was taken out from femoral vein by a snare-catheter apparatus. Another patient developed CS dissection and small pericardium perfusion during LV lead implantation (upgrade to BiV-ICD) post successful lead extraction and patient discharged without complication. CONCLUSION: Our single center experience suggests that Excimer Laser Sheath Lead Removal System is safe and effective in extracting pacemaker and ICD leads. PMID- 21624316 TI - [A case report of Kearns-Sayre syndrome]. PMID- 21624317 TI - [Two cases of giant Epsilon wave in patients with arrhythmogenic right ventricular dysplasia]. PMID- 21624318 TI - [A case report of primary amyloidosis manifested by bilateral lower extremity edema and dyspnea]. PMID- 21624319 TI - [Evaluation on stepwise ablation approach for long-standing persistent atrial fibrillation]. PMID- 21624320 TI - [Role of circulating microRNA on clinical diagnosis of cardiovascular diseases]. PMID- 21624321 TI - [Progress of cohort study and its inspiration to China Health and Nutrition Survey]. PMID- 21624322 TI - [Municipal solid waste incinerator and cancer incidences among the residents in the vicinity]. PMID- 21624323 TI - [Milk consumption and its changing trend of Chinese adult aged 18 - 44 in nine provinces (autonomous region) from 1991 to 2006]. AB - OBJECTIVE: To investigate the age, gender and region specified milk consumption and its changing trends of Chinese 18 - 44 years old adults in 9 provinces (autonomous region) from 1991 to 2006. METHODS: The data of 18 - 44 aged old adults investigated in Chinese Health and Nutrition Survey from 1991 to 2006 in 9 provinces (autonomous region) were utilized. The numbers of the subjects selected in 1991, 1993, 1997, 2000, 2004 and 2006 were 5593, 5491, 5334, 4188 and 3908, respectively. Changes of the percentage of milk consumption, average intake of daily milk and dietary calcium intake were analyzed across different groups of gender, income levels, and areas. RESULTS: There was an increasing trend in the percentage of milk consumption and daily intake of milk from 1991 to 2006. The average percentage of Chinese adult milk consumption went up from 2.2% (123/5593) in 1991 to 7.6% (297/3908) in 2006 (Z = -22.16, P < 0.01). Specifically, urban adult and rural adult showed significant increase in the percentages from 9.7% (80/823) to 23.7% (112/472) (Z = -12.81, P < 0.01) and 0.1% (2/2937) to 1.8% (36/1987) (Z = -9.21, P < 0.01) respectively. Percentages of adult who ever drank milk at top income level and bottom income level increased from 4.7% (95/2006) to 14.6% (200/1373) (Z = -17.36, P < 0.01) and from 0.1% (2/1748) to 2.4% (28/1171) (Z = -9.58, P < 0.01), respectively. Meanwhile, the average daily intake of milk increased from (3.6 +/- 24.6) g/d in 1991 to (11.8 +/- 51.4) g/d in 2006 (F = 67.07, P < 0.01). The daily milk consumption among top income level adults and bottom income level adults increased from (8.5 +/- 50.9) g/d to (23.1 +/- 71.5) g/d and from (0.2 +/- 6.5) g/d to (3.3 +/- 23.5) g/d (F = 13.90, P < 0.01), respectively. From 1991 to 2006, milk consumers significantly promoted their percentage of achieving the Chinese dietary reference intake on calcium (800 mg/d) from 6.5% (8/123) to 12.8% (38/297) (Z = -1.67, P < 0.05), whereas this percentage increased from 1.6% (38/5470) to 3.6% (130/3611) among milk non consumers (Z = -5.66, P < 0.01). CONCLUSION: Chinese adult milk intake still remains in quite low level, and the situation of milk consumption among rural adults requires further promotion. PMID- 21624324 TI - [The status of dietary sodium intake of Chinese population in nine provinces (autonomous region) from 1991 to 2006]. AB - OBJECTIVE: To analyze the status of dietary sodium intake of 18 - 65 years old Chinese people in nine provinces (autonomous region) in 1991 and 2006. METHODS: In 1991 and 2006, China Health and Nutrition Survey was carried out in nine provinces, including Liaoning, Heilongjiang, Shandong, Jiangsu, Henan, Hunan, Hubei, Guangxi and Guizhou, Subjects were selected by multi-stage stratified cluster random sampling method from 18 - 65 years old people and 8235 subjects in 1991 and 7788 subjects in 2006. Sodium intake analysis was conducted across gender, regions and rural(urban) residences. RESULTS: On average, sodium intake decreased from (6750.6 +/- 3771.5) mg/d in 1991 to (4949.7 +/- 2829.4) mg/d in 2006 (Z = -35.5, P < 0.01) among the 18 - 65 years old people. In 1991 and 2006, the main sources of dietary sodium were salt, soy sauce and vegetable. In 1991, the proportion were 79.6% ((5375.6 +/- 3532) mg/d), 8.7% ((586.7 +/- 721.7) mg/d) and 7.5% ((506.9 +/- 906.1) mg/d), respectively. In 2006, the proportion were 71.5% ((3539.8 +/- 2545) mg/d), 8.3% ((410.7 +/- 547.9) mg/d) and 6.2% ((304.6 +/ 610.7) mg/d), respectively. CONCLUSIONS: Among 18 - 65 years old residents, sodium intake in 2006 was significantly lower than that in 1991, but still far beyond the 2200 mg/d of adequate intake level (2200 mg/d). The major source of sodium intake in Chinese diet was condiments. PMID- 21624325 TI - [Milk consumption and effects on dietary nutrients and growth status among Chinese children in nine provinces (autonomous region) from 1991 to 2006]. AB - OBJECTIVE: To investigate milk consumption and effects on dietary nutrients and growth status among 7 - 17 years-old Chinese children in 9 provinces (autonomous region) from 1991 to 2006. METHODS: Data was collected in 1991, 1993, 1997, 2000, 2004 and 2006 China Health and Nutrition Survey, and 11 691 subjects aged 7 to 17 years old were included. The subjects were divided into the following groups according to milk consumption per day, 0 (no milk drinking), 1 - 99, 100 - 199, 200 - 299 and >= 300 g/d. Variables from different years were compared, such as milk categories, energy, protein, calcium intake, body mass index (BMI), as well as that dietary nutrients, height and weight among different milk consumption groups. RESULTS: From 1991 to 2006, milk drinking rate and milk consumption among Chinese children had great improvement: milk drinking rate reached to 14.1% (161/1145) in 2006, while that was 3.0% (73/2441) in 1991; milk consumption reached to (26.7 +/- 85.0) g/d in 2006, which was 6.8 times of that in 1991 (3.9 +/- 31.9) g/d (chi(2) = 474.5, P < 0.01). BMI increased from (17.3 +/- 2.9) kg/m(2) in 1991 to (18.5 +/- 6.0) kg/m(2) in 2006 (chi(2) = 123.5, P < 0.01). Meanwhile, energy intake decreased from (8653.3 +/- 2789.2) kJ/d in 1991 to (8058.3 +/- 2866.6) kJ/d in 2006 (chi(2) = 72.6, P < 0.01); protein intake decreased from (63.0 +/- 22.8) g/d in 1991 to (57.3 +/- 23.1) g/d in 2006 (chi(2) = 71.9, P < 0.01). Average height of 0 (no milk drinking), 1 - 99, 100 - 199, 200 - 299 and >= 300 g/d groups were (142.9 +/- 16.9), (146.9 +/- 18.8), (147.6 +/- 16.2), (148.8 +/- 16.0), (149.9 +/- 15.3) cm, respectively (chi(2) = 29.4, P < 0.01); average weight were (37.1 +/- 13.2), (40.3 +/- 13.9), (41.3 +/- 16.9), (41.7 +/- 14.6), (41.4 +/- 12.2) kg, respectively (chi(2) = 25.7, P < 0.01). CONCLUSION: In the period of 1991 to 2006, milk drinking rate and milk consumption has improved greatly. Main nutrient intake, height and weight increased with milk consumption. PMID- 21624326 TI - [Trends of the dietary fiber intake among Chinese aged 18 - 45 in nine provinces (autonomous region) from 1989 to 2006]. AB - OBJECTIVE: To analyze the status and trend of dietary fiber intake and its food sources among Chinese residents aged 18 - 45 years old in 9 provinces (autonomous region), from 1989 to 2006. METHODS: Twenty-four-hour dietary recall data of three consecutive days from China Health and Nutrition Survey (1989, 1991, 1993, 1997, 2000, 2004 and 2006) were used. And the 18 - 45 years old subjects with complete information on dietary survey were included. The sample size of these 7 surveys were 5597, 5596, 5323, 5485, 5308, 4144 and 3889, respectively. The trends of dietary fiber intake and food sources by time and sex were studied. RESULTS: From 1989 to 2000, the total of daily dietary fiber intake of 18 - 45 years old healthy residents decreased from 15.1 g/d to 11.6 g/d, insoluble dietary fiber intake decreased from 22.6 g/d to 17.8 g/d. In villages, from 1989 to 2000, the insoluble dietary fiber intake decreased 5.1 g/d in males and 5.2 g/d in females(13.2, 11.8 g in 2000, respectively;18.3, 17.0 g in 1989, respectively). In 2006, the insoluble dietary intake was 12.6 g/d in males and 11.2 g/d in females in villages. In cities, from 1989 to 2006, the insoluble dietary fiber intake was 9.4 - 11.8 g/d in males and 8.3 - 10.8 g/d in females. The total dietary fiber intake was 15.7 - 17.6 g/d in males and 13.5 - 16.4 g/d in females. The difference of insoluble and total dietary fiber intake between city and village was decreasing. In 2006, 70.6% (8.4/11.9) insoluble dietary fiber came from vegetables ((4.1 +/- 3.1) g/d), wheat flour and its products ((2.6 +/- 3.6) g/d), rice and its products ((1.7 +/- 1.3) g/d). CONCLUSION: Dietary fiber intake among Chinese aged 18 - 45 in 9 provinces (autonomous region) decreased from 1989 to 2006. Vegetables, wheat flour, rice and their products were the major food sources of insoluble dietary fiber. PMID- 21624327 TI - [Status and trend of alcohol consumption among adults in nine provinces (antonomous region) of China from 1993 to 2006]. AB - OBJECTIVE: To understand the status and trend of alcohol consumption among adult people living in 9 provinces (autonomous region) of China from 1993 to 2006. METHODS: The data was collected at the China Health and Nutrition Survey in 1993, 1997, 2000, 2004 and 2006, the subjects with complete drinking data of adult residents were involved. The sample of subjects were 7994, 8625, 9283, 9780 and 9746 among 5 round surveys respectively. Alcohol intake, drinking frequency, drinking type, drinking amount and trends of changing was analyzed. RESULTS: The average drinking rates were 60.2% (13 201/21 919) and 9.7% (2269/23 509) among male and female respectively. The average alcohol intake was (32.2 +/- 35.6) g in male and (14.0 +/- 19.4) g in female drinkers. Male drinkers with primary and secondary schools or below education had the lowest level of drinking rate, which was 59.7% (985/1651), 58.7% (941/1604), 57.0% (889/1560), 55.4% (885/1599) and 53.1% (812/1529) in the 5 round surveys respectively. However, the average daily alcohol intake was the highest, which was (31.1 +/- 38.2) g, (39.8 +/- 42.1) g, (38.9 +/- 41.3) g and (38.4 +/- 43.3) g at 1997, 2000, 2004, 2006 respectively. The male drinkers with college or above education had the lowest alcohol intake, which was (21.9 +/- 24.5) g, (28.8 +/- 30.9) g, (24.6 +/- 22.0) g and (25.0 +/- 26.4) g in 1997, 2000, 2004 and 2006 respectively. Female drinkers with college or above education had the highest drinking rate, which was 21.0% (13/62), 24.5% (26/106), 16.6% (27/163), 19.7% (38/193) and 18.8% (46/245) at 5 round surveys respectively. But their average daily alcohol intake was lowest, which was (4.7 +/- 3.3) g, (12.5 +/- 31.9) g, (9.9 +/- 18.7) g and (8.2 +/- 8.5) g at 1997, 2000, 2004, 2006 respectively. And the average daily alcohol intake was the highest among female drinkers with primary and secondary schools or below education, which was (13.4 +/- 16.9) g, (16.3 +/- 22.4) g, (19.9 +/- 26.1) g and (16.4 +/- 18.0) g respectively. According to the regional distribution, the highest drinking rate of female drinker was occurred at urban female, which was 17.4% (110/631), 19.9% (147/740), 17.4% (129/741), 14.4% (117/814) and 11.3% (88/781) at 5 round surveys. Drinking every day was most popular in male drinkers, the proportion was 32.6% (3533/10 844). Drinking 1 to 2 times per week, 1 to 2 times per month, no more than 1 time per month was most popular in female drinkers, the proportion was 23.0% (410/1786), 24.6% (440/1786) and 21.2% (379/1786) respectively. Male drinkers liked liquor more (drinking rate was 81.8%, 8875/10 844). The wine drinking rate of female (22.3%, 399/1786) is far higher than that of male's (9.0%, 976/10 844) (chi(2) = 280.4, P < 0.01). The daily alcohol intake equal to or greater than 25 g of male drinkers was 42.0% (4058/9654), and the daily alcohol intake equal to or greater than 15 g of female drinkers was 25.3% (349/1378). CONCLUSION: The drinking rate of adults among 9 provinces (autonomous region) was higher than that of national average level. The drinking rate and the average alcohol consumption among male and female was significantly different. Same differences was observed among education level and different regions. PMID- 21624328 TI - [Food consumption trend of Chinese adults in nine provinces (autonomous region) from 1989 to 2006]. AB - OBJECTIVE: To analyze the trends of food consumption among Chinese residents aged from 18 to 59 years old in 9 provinces (autonomous region) from 1989 to 2006. METHODS: The paper was based on data collected in 1991, 1993, 1997, 2000, 2004 and 2006 China Health and Nutrition Survey of Chinese residents of 24 h in three consecutive day dietary recall data. The healthy residents aged 18 - 59 years with twenty-four-hour dietary recall data were selected as study subjects. The sample sizes of these seven surveys were 5766, 7296, 7019, 7599, 8143, 7168 and 6827, respectively. The trends of their food consumption in each year were analyzed through Kruskal-wallis method. RESULTS: The intake of cereals decreased significantly from (528.5 +/- 192.9) g/d in 1989 to (427.7 +/- 173.8) g/d in 2006 (chi(2) = 2833.13, P < 0.01). The minor variation of soybean food intake was observed. The average intake was (22.3 +/- 39.4) g/d in 1989 and (18.9 +/- 30.5) g/d in 2006 which was still at a low level. The trend of vegetables intake showed a small decrease. It was (374.5 +/- 205.9) g/d in males and (346.4 +/- 187.6) g/d in females in 2006, respectively. Both quantity and rate of fruits consumption showed an increase trend. Its consumption rate reached at 22.5% (741/3296) in males and 27.7% (1004/3619) in females in 2006, respectively. The average intake increased from (122.4 +/- 126.9) g/d in 1989 to (228.5 +/- 267.3) g/d in 2006 (chi(2) = 539.66, P < 0.01). At the same time, the daily intake of meat and egg showed a large increase, the average meat intake increased from (97.0 +/- 111.5) g/d in males and (80.2 +/- 97.0) g/d in females in 1989 to (141.5 +/- 125.2) g/d in males and (118.4 +/- 111.8) g/d in females in 2006, respectively. The oil consumption showed a significant increase. The average intake increased from (30.4 +/- 37.9) g/d in 1989 to (42.8 +/- 32.3) g/d in 2006 (chi(2) = 1646.94, P < 0.01). CONCLUSIONS: There have been noticeable changes in the Chinese dietary pattern in the past 17 years. Some changes would be helpful for nutritional improvement in Chinese people, but it also caused some adverse effects on health because of improper food consumption. PMID- 21624329 TI - [Effects of soybean isoflavone on liver lipid metabolism in nonalcoholic fatty liver rats]. AB - OBJECTIVE: To study the effects of soybean isoflavone on liver lipid, serum lipid, antioxidant index and hepatic lipid metabolism associated factors in nonalcoholic fatty liver rats. METHODS: Thirty-six male rats (SD) were randomly divided into four groups by weight: normal control group, nonalcoholic fatty liver disease (NAFLD) model control group, low-dose isoflavone treatment group (10 mg/kg) and high-dose isoflavone group (20 mg/kg), 9 rats in each group. Normal control rats were fed with D12450B (10% fat energy), model control and isoflavone intervention rats were fed with D12492 (60% fat energy). Twelve weeks later, liver lipid, serum lipid and antioxidant index were observed. Liver sterol regulatory element binding protein-1c (SREBP-1c), fatty acid synthase (FAS) and peroxisome proliferators activated receptor alpha (PPAR alpha) were detected by western blotting. RESULTS: Liver triglyceride (TG) in normal control group, NAFLD model control group, low-dose isoflavone group and high-dose isoflavone group were (8.11 +/- 4.13), (57.06 +/- 16.95), (31.26 +/- 10.48), (31.38 +/- 13.25) mmol/mg protein, respectively (F = 22.569, P < 0.01); liver free fatty acid (FFA) were (0.030 +/- 0.007), (0.042 +/- 0.009), (0.038 +/- 0.009), (0.032 +/- 0.005) umol/mg protein, respectively (F = 4.857, P < 0.01); liver superoxide dismutase (SOD) activity were (502.29 +/- 23.71), (201.83 +/- 16.99), (228.93 +/- 21.71), (238.08 +/- 15.96) U/mg protein, respectively (F = 9.555, P < 0.01); liver malondialdehyde (MDA) were (1.29 +/- 0.29), (2.85 +/- 0.73), (2.07 +/- 0.49), (2.03 +/- 0.37) nmol/mg protein, respectively (F = 13.449, P < 0.01); SREBP-1c protein expression were 0.45 +/- 0.16, 1.42 +/- 0.30, 1.02 +/- 0.31, 0.47 +/- 0.27, respectively (F = 24.515, P < 0.01); FAS protein expression were 0.27 +/- 0.08, 1.97 +/- 0.47, 1.35 +/- 0.30, 0.49 +/- 0.12, respectively (F = 60.361, P < 0.01); PPARalpha protein expression were 2.03 +/- 0.56, 0.41 +/- 0.17, 0.81 +/- 0.27, 0.66 +/- 0.16, respectively (F = 37.97, P < 0.01). CONCLUSION: Soy isoflavone can reduce the hepatic lipid deposition and increase antioxidant capacity, the mechanism may be related to inhibition of SREBP-1c and activation of PPARalpha expression in liver. PMID- 21624330 TI - [Effects of lanthanum chloride on the expression of immediate early genes in the hippocampus of rats]. AB - OBJECTIVE: To study influence of lanthanum chloride (LaCl(3)) on the expression of immediate early genes (IEGs) including c-jun, early growth response gene 1 (Egr1) and activity-regulated cytoskeletal gene (Arc) in the hippocampus of rats, and discuss the mechanism of LaCl(3) undermining learning and memory capability. METHODS: Forty female Wistar adult rats were divided into control group, low LaCl(3)-contaminated group (0.25%), medium LaCl(3)-contaminated group (0.50%), and high LaCl(3)-contaminated group (1.00%) by randomized design. Each group had ten female rats along with five male rats and mated by the ratio of 2:1. The amounts of pups in the above four groups were 80, 83, 78 and 75 separately. The pups in respective group were La-dyed by lactation, and then the pups in LaCl(3) contaminated groups drank 0.25%, 0.50% and 1.00% LaCl(3) separately for one month. Learning and memory capability of pups were measured in jumping stairs experiment. Hippocampal lanthanum content was determined by inductively coupled plasma mass spectrometry (ICP-MS). Hippocampal c-jun, Egr1 and Arc mRNA expression was detected by RT-PCR, and corresponding protein expression was measured by Western blotting method. RESULTS: In the jumping stairs experiment, pups in 0.25%, 0.50% and 1.00% LaCl(3)-contaminated groups respectively made (1.75 +/- 0.71), (2.38 +/- 0.92) and (3.00 +/- 0.76) mistakes; significantly higher than control group (1.25 +/- 0.46) (q values were 4.386, 6.793, P < 0.05). However, the incubation period of 0.25%, 0.50% and 1.00% LaCl(3)-contaminated groups were (174.13 +/- 33.72), (139.25 +/- 45.83) and (75.50 +/- 18.56) respectively, which were all significantly lower than that of control group (206.75 +/- 20.47) (q values were 2.958, 6.121, 11.902, P < 0.05). Hippocampal c jun mRNA expression were (0.89 +/- 0.08), (0.77 +/- 0.12), (0.58 +/- 0.14) and (0.29 +/- 0.10); while the c-jun protein expression were (0.72 +/- 0.13), (0.64 +/- 0.11), (0.43 +/- 0.11) and (0.31 +/- 0.14), and the Egr1 mRNA expression were (0.78 +/- 0.09), (0.61 +/- 0.13), (0.53 +/- 0.10) and (0.22 +/- 0.08), Egr1 protein expression were (0.65 +/- 0.18), (0.40 +/- 0.15), (0.32 +/- 0.13) and (0.14 +/- 0.09) in 0.25%, 0.50% and 1.00% LaCl(3)-contaminated groups; and all of which presented a dose-effect relationship that the correlation coefficients of these parameters with dose were -0.900 (t = 11.309, P = 0.000), -0.969 (t = 7.058, P = 0.000), -0.898 (t = 11.179, P = 0.000) and -0.962 (t = 6.739, P = 0.000). CONCLUSION: LaCl(3) undermines the learning and memory capability of rats, which is possibly related to lower expression of c-jun and Egr1 gene and protein induced by lanthanum in hippocampus. PMID- 21624331 TI - [A correlative study on Bisphenol A and recurrent spontaneous abortion]. AB - OBJECTIVE: This study was to investigate the association of Bisphenol A and unexplained recurrent spontaneous abortion. METHODS: A hospital-based 1:1 matched case-control study was conducted. Sixty patients with unexplained recurrent abortion were included. Each case was matched with one normal control by age (+/- 2 years), living district and the same gestational age. The levels of Bisphenol A in urine for 60 cases and 60 controls were detected using high performance liquid chromatography after fluorescent derivatization. The levels of urinary Bisphenol A in case was compared with that in control in education levels, occupation, smoking history. Data was analyzed by means of Wilcoxon-test, Student-Newman Keuls after rank transform, univariate and multivariate conditional Logistic regression analysis. The software used was SAS 9.1.3. RESULTS: The values of urinary Bisphenol A in cases and controls were (0.10 +/- 0.21) ug/ml, (0.03 +/- 0.08) ug/ml, respectively. The level of urinary Bisphenol A in cases was significantly higher than that in controls (Z = 3.988, P < 0.0001). The urinary Bisphenol A levels in cases were significant higher than those in controls from senior middle school (the levels were 0.10, 0.06 ug/ml respectively, Z = 1.996, P = 0.0459), college (the levels were 0.14, 0.03 ug/ml respectively, Z = 2.586, P = 0.0097), workers or farmers (the levels were 0.08, 0.03 ug/ml respectively, Z = 2.265, P = 0.0235), businessmen (the levels were 0.10, 0.03 ug/ml respectively, Z = 2.544, P = 0.0109), and no passive smokers (the levels were 0.09, 0.03 ug/ml respectively, Z = 3.767, P = 0.0002). After adjustment by age, body mass index, marital status during pregnancy and other factors, compared to Bisphenol A below 0.06 ug/ml, the adjusted OR was 4.03 (1.67 - 9.74) for Bisphenol A levels between 0.06 ug/ml and 0.20 ug/ml, and was 5.46 (1.95 - 15.27) for Bisphenol A over 0.20 ug/ml. The risk of unexplained recurrent spontaneous abortion increased progressively with the growth of urinary Bisphenol A levels (chi(2) = 13.042, trend test P = 0.0003). There were significant differences on Bisphenol A among controls, two abortions, and three or more abortions (the levels were 0.03 ug/ml, 0.09 ug/ml, 0.21 ug/ml respectively, F = 9.04, P = 0.0002). CONCLUSION: Exposure to Bisphenol A may be associated with unexplained recurrent spontaneous abortion. PMID- 21624332 TI - [The relationship between secular trend of road traffic injuries and gross domestic product per capita in China]. AB - OBJECTIVE: To explore the relationship between secular trend of road traffic injuries (RTI) and gross domestic product (GDP) per capita in China. METHODS: Statistical description was used in the data about cases, injuries, deaths, mileage mortality and 10 million population mortality from 1970 to 2009. Cluster analysis was used to classify the 31 provinces, municipalities and autonomous regions in China. Ecological study was used to explore the relationship between RTI and GDP per capita. RESULTS: There were three stages of RTI in China. It grew rapidly in 1970 - 2002 (from 1.16 to 8.52 per 10 million population), kept steady in 2003 - 2004 (from 8.08 to 8.24 per 10 million population), and decreased obviously in 2005 - 2009 (from 7.55 to 5.08 per 10 million population). The ecological study showed that the population mortality of RTI rose along with the GDP per capita's growth. When the GDP per capita reached to 14 053 yuan (equivalent to 1716 US dollar, in 2005), the mortality began to decrease obviously, the average annual decreasing rate was 10.16%(8.14% - 10.52%)in the following five years. According to the GDP per capita during the period of 1999 - 2009, the 31 provinces, municipalities and autonomous regions in China were divided into three categories of region. The curves of population mortality of RTI and GDP per capita in different category possessed the same ecological trend. That was the population mortality early rose and then fell along with the GDP per capita's growth. All of they started to decrease obviously in 2005. The GDP per capita among three categories of region was different (45 281 yuan, 22 243 yuan and 10 475 yuan respectively) in the same period. CONCLUSION: In the early stage of economic development, the mortality of RTI increased along with the economic development. When the economic development reached a certain level, the mortality decreased along with the GDP per capita's growth. PMID- 21624333 TI - [A follow-up study on the post-traumatic stress disorders among middle school students in Wenchuan earthquake region]. AB - OBJECTIVE: This study was to identify the post-traumatic stress disorder (PTSD) changes and the relative risk factors within one year after Wenchuan earthquake among middle school students in the disaster area. METHODS: A total of 1966 students from 3 schools in Wenchuan earthquake region were selected as the target population. For each student, personal basic information and standard psychological scale (PCL-C, PSSS) were investigated by a self-administrated questionnaire in the 3rd, the 6th, the 9th and the 12th month after the earthquake, respectively. PTSD trends over the time and the associated risk factors were analyzed through the establishment of multi-level random coefficient model. RESULTS: There were 1677 middle school students fully participated in the PTSD follow-up study by turning in the valid questionnaires. The averaged scores of PTSD at the time of the 3rd, the 6th, the 9th and the 12th month after the earthquake were 35.14 +/- 11.08, 32.90 +/- 11.03, 30.67 +/- 11.28 and 29.75 +/- 11.22, respectively. Meanwhile, the general incidences of PTSD were 36.6% (613/1677), 30.7% (515/1677), 24.8% (416/1677)and 22.2% (373/1677), respectively. The median score of perceived social support system was 60.00 and the general incidences of PSS was 17.20% (289/1677). The PTSD scores for the students had a decreasing trend during the period of our observation (beta(time) = -1.879, chi(2) = 47.03, P < 0.05). The averaged scores for boys for the 4 follow-up studies were 33.71, 31.61, 29.66, 28.83; for girls were 36.33, 33.98, 31.51, 30.52; for junior school students were 35.46, 33.28, 30.18, 29.22; for senior school students were 34.89, 32.62, 31.04, 30.15. Moreover, two factors, gender and grade, were related with the decreasing trend (the trend for girls and senior school students was sharper than that for boys and junior school students) (beta(gender-time) = -0.354, chi(2) = 4.83, P < 0.05; beta(grade-time) = 0.622, chi(2) = 11.30, P < 0.05). CONCLUSIONS: The prevalence of PTSD was high. Meanwhile, there was a trend of self-recovery for adolescent's PTSD during the first year of post-earthquake, but boys and junior school students recovered more slowly. PMID- 21624334 TI - Estrogen regulates the expression and activity of epithelial sodium channel in mouse osteoblasts. AB - Estrogen plays an important role in bone metabolism and only high dose can stimulate osteoblast bone formation. However, the underlying mechanisms have not been elucidated. The epithelial sodium channel (ENaC) is a key pathway for sodium transport in epithelia, vascular endothelium, and other tissues; although the expressions of alpha and gamma ENaC mRNA were found in osteoblasts, the regulation of ENaC by estrogen in osteoblasts has not been studied. Our recent data confirmed the ENaC expression in mouse primary osteoblasts by immunocytofluorescence, RT-PCR, western blot, and patch clamp. Furthermore, we found estrogen (10(-5)M) increased the expression of alpha and gamma ENaC subunits at both the mRNA and protein levels in osteoblasts. On the other hand, 17beta estradiol (20 nM) increased inward Na+ currents which were inhibited by amiloride. The estrogen dose used in patch clamp is much lower than those of mRNA and protein analysis, which means single cell ENaC electrophoretic mobility is much more sensitive to estrogen than the mRNA and protein production by estrogen stimulation. Our results suggest that estrogen regulates expression and function of ENaC in osteoblasts may provide a new clue that the mechanism of high dose of estrogen influence osteoblast bone formation via ENaC activity. PMID- 21624335 TI - Cytotoxic effects of Argentinean plant extracts on tumour and normal cell lines. AB - In the search for possible new anti-cancer agents, we investigated the effects of 75 aqueous and methanol extracts from 41 Argentinean plant species. The effect in cell growth was evaluated in the LM2 mammary adenocarcinoma cells. In a second stage, the highly active selected extracts were assayed in 3 other tumour cell lines: melanoma B16, bladder MB49 and lung A549; and 3 normal cell lines: mammary Hb4a and keratinocytes PAM212 and HaCat. Eight methanol extracts were found to be highly cytotoxic: Collaea argentina leaf, Iochroma australe leaf, Ipomoea bonariensis flower, Jacaranda mimosifolia flower, Solanum amygdalifolium flower, Solanum chacoense leaf, Solanum sisymbriifolium flower and Solanum verbascifolium flower. However, extract inhibition on cell growth was highly dependent on cell type. In general, except for the highly resistant cell lines, the inhibitory concentrations 50% were in the range of 10-150 MUg/ml The eight extracts highly inhibited cell growth in a concentration-dependent manner, and in general the methanol extracts were always more active than the aqueous. Murine cells appear to be more sensitive than human cells to the cytotoxic action of the plant extracts. The human melanoma B16 line was the most resistant to four of the extracts. In terms of selectivity, S. verbascifolium was the species which showed most selectivity for tumour cells. Overall, this is one of the first studies focusing on southern South American native plants and their biological effects. Since some species of 5 genera analyzed have been reported to possess different degrees of alkaloid content, we examined microtubule structures after extract treatments. The eight extracts induced destabilization, condensation and aggregation of microtubules in LM2 cells, although no depolarization, typical of Vinca alkaloids damage was observed. In a near future, antitumour activity of purified fractions of the extracts administered at non-toxic doses will be assayed in transplantable murine tumour models. PMID- 21624336 TI - LevRad software as a tool to learn how to proceed with an evaluation of barriers. AB - We developed the software LevRad with the objective of teaching how to proceed in an analysis of barriers shielding against x-rays to minimize the contact of the professional or the student with x-rays and also to prevent wearing out of the x ray equipment. Some tests of the software were made, and preliminary results indicate that LevRad is efficient as a complementary tool for the development of professionals related to diagnostic radiology. In the case of education, an advantage is gained when the beginner uses the software before his or her first contact with x-ray equipment in locu. The software introduces a basic knowledge about evaluation of barriers, prevents wearing out of the x-ray tube, reinforces teaching of evaluation of barriers, and reduces the collective effective dose by avoiding unnecessary exposures when possible. PMID- 21624337 TI - [Aspects and relevant relationship in the nursing workload conceptualization: literature review]. AB - The authors describe how the definition of work-load in nursing is susceptible to many variables such as the number of patients cared for and their specific needs, the mix of skills required and the activities performed by nurses not strictly pertinent to care. These features make it difficult to create a global method for allocation of nursing resources . The authors have reviewed the literature from 2000 to 2009 in an attempt to identify the relationships between the concepts at the basis of the definition of work-load in nursing. The concepts of complexity and intensity of care are analyzed together with the various systems used for allocation of resources in different countries. Their results show that work-load is an important element of nursing organization and can influence the outcome of care. PMID- 21624338 TI - [Creating a nursing record system: implementation of assessment forms]. PMID- 21624339 TI - [A new resource for the bibliography research: project experience ILISI(r) Index of Italian Literature on Nursing Sciences]. AB - Since July 2008 the ILISI (Index of Italian Literature on Nursing Sciences) elaborated by the IPASVI nursing college of Rome has been available on-line at the page http://www.ipasvi.roma.it/ita/ILISI/ . The aim of this is to make Italian nursing literature more available and to favor bibliographic research. About 3000 articles and 30 nursing journals are available : the necessary software is open source (free) and has been adapted to allow searches by author, topic or word content. Indexation has been carried out by a group of volunteer nurses using a Thesaurus created by the project group. This article describes the aims of the project , how it has been created , the resources employed and the potential of the database. Use of the latter is on the increase: in fact, during the first 12 months of availability , the number of consultations reached 9000. PMID- 21624340 TI - [Stress and nursing : study to evaluation the level of satisfaction in nurses]. AB - Not unfrequently stress in nurses is related to work organization. The literature shows that the nursing profession has a high risk of psychological stress. Stress is one of the factors that contributes to inefficiency, increasing staff turnover and sick leave, and reduces the quality and quantity of care , affecting health costs and diminishing work satisfaction. This article presents the results of a study performed to evaluate 34 variables, constituting the Nursing Stress Scale (NSS) , and their level in the nursing staff of a Rome specialistic hospital. Of the 80 nurses who received the questionaire , 49 responded, a sufficient number to make the results of the study significant. The authors illustrate the methods used together with some socio-demographic considerations and how some of these can be correlated to the 7 factors adopted for multi-variate analysis. The results obtained were concordant with those described in the literature, confirming that poor organization and conflictual or ambiguous work roles negatively affect well-being and that nursing directors should bear this in mind when aiming to create a harmonious work environment. PMID- 21624341 TI - [Sexual dimorphism and facial cavities: a 3D imaging volumetric study]. AB - INTRODUCTION: The aim of this study was to investigate the sexual dimorphism in facial cavity's volumes of untreated young adults. METHODS: Sixty subjects, ages between 18 to 30 years, were selected and divided in two groups according to the gender. Volumes of orbital, nasal, sinusal and buccal cavities were measured from CT scans with AMIRA((r))software. The statistical studies were performed using Kolmogorov-Smirnov test for normality followed by Student "t" test (p < 0.05). RESULTS: The gender comparison reflected statistically significant differences between the volumes of orbital, nasal, sinusal and buccal cavities, volumes being larger in males. Sexual dimorphism did not appear in the ratio of each cavity volume on the total facial volume. CONCLUSION: Absolute volume of facial cavities are larger in males, but no sexual dimorphism is detected comparing volume ratios. A difference in size, but not in shape, between genders might exist. PMID- 21624342 TI - [A classification of cranio-facial syndromes]. AB - How to manage craniofacial malformative cases? It seems to be very difficult, especially in orthodontics because of the lack of consensus. The authors' aim is to propose a physiopathologic classification of these craniofacial syndromes in order to simplify the medical practice when we meet these patients. More than fifty cases are actually treated and followed in our hospital; we have described all of these cases before to choose the most representative in each category. These syndromes are classified in four categories, organ abnormalities of one or many functional matrix, localized abnormalities of the anatomical structures, general abnormalities of the connective tissue, mixed syndromes. PMID- 21624343 TI - A robust RNA integrity-preserving staining protocol for laser capture microdissection of endometrial cancer tissue. AB - Laser capture microdissection of frozen tissue sections allows homogeneous cell populations to be isolated for expression profiling. However, this requires striking a balance between retaining adequate morphology for accurate microdissection and maintaining RNA integrity. Various staining protocols were applied to frozen endometrial carcinoma tissue sections. Although alcohol-based methods were superior to aqueous stains for maintaining RNA integrity, they suffered from irreproducible staining intensity. We developed a modified alcohol based, buffered cresyl violet staining protocol that provides reproducible staining with minimal RNA degradation suitable for tissues with moderate to high levels of intrinsic RNase activity. PMID- 21624344 TI - A cost-effective method for simultaneous homo-oligomeric size determination and monodispersity conditions for membrane proteins. AB - The use of blue native polyacrylamide gel electrophoresis (BN-PAGE) has been reported in the literature to retain both water-soluble and membrane protein complexes in their native hetero-oligomeric state and to determine the molecular weight of membrane proteins. However, membrane proteins show abnormal mobility when compared with water-soluble markers. Although one could use membrane proteins as markers or apply a conversion factor to the observed molecular weight to account for the bound Coomassie blue dye, when one just wants to assess homo oligomeric size, these methods appear to be too time-consuming or might not be generally applicable. Here, during detergent screening studies to identify the best detergent for achieving a monodisperse sample, we observed that under certain conditions membrane proteins tend to form ladders of increasing oligomeric size. Although the ladders themselves contain no indication of which band represents the correct oligomeric size, they provide a scale that can be compared with a single band, representing the native homo-oligomeric size, obtained in other conditions of the screen. We show that this approach works for three membrane proteins: CorA (42 kDa), aquaporin Z (25 kDa), and small hydrophobic (SH) protein from respiratory syncytial virus (8 kDa). In addition, polydispersity results and identification of the most suitable detergent correlate optimally not only with size exclusion chromatography (SEC) but also with results from sedimentation velocity and equilibrium experiments. Because it involves minute quantities of sample and detergent, this method can be used in high-throughput approaches as a low-cost technique. PMID- 21624345 TI - Evaluation of anti-depressant-like activity of linezolid, an oxazolidinone class derivative - an investigation using behavioral tests battery of depression. AB - Linezolid, an oxazolidinone class derivative is a reversible and nonselective inhibitor of monoamine oxidase (MAO), predominantly for MAO-A type. MAO-A is a key enzyme regulating the catabolism of catecholamine neurotransmitters in the brain. It is well known that the catecholaminergic neuronal systems are associated with depression and inhibition of MAO-A level in the brain could be used to treat depression. Hence, the objective of this study was to evaluate the anti-depressant-like effect of linezolid, a MAO-A inhibitor in the animal models of depression. In the present study, linezolid (10 & 20mg/kg, i.p.), exhibited anti-depressant-like effects in forced swim test (FST) and tail suspension test (TST) in mice without influencing the baseline locomotion. Moreover, linezolid (10 & 20mg/kg, i.p.), potentiated the 5-hydroxytryptophan (5-HTP)-induced head twitch responses in mice and antagonized the reserpine-induced hypothermia in rats. In conclusion, the behavioral investigation revealed the anti-depressant like effect of linezolid in rodent's behavioral model. PMID- 21624346 TI - Keratan sulfate and related murine glycosylation can suppress murine cartilage damage in vitro and in vivo. AB - Keratan sulfate (KS) proteoglycan side chains are abundant in the human cartilage matrix, but these chains have been said to be absent in murine skeletal tissues. We previously showed that KS suppresses cartilage damage and ameliorates inflammation in mice arthritis model. Because mice deficient of N acetylglucosamine 6-O-sulfotransferase-1 (GlcNAc6ST-1) (KS biosynthesis enzyme) are now available, we decided to do further examinations. We examined, in culture, the difference between GlcNAc6ST-1(-/-) and wild-type (WT) mice for interleukin (IL)-1alpha-induced glycosaminoglycan (GAG) release from the articular cartilage. Arthritis was induced by intravenous administration of an anti-type II collagen antibody cocktail and subsequent intraperitoneal injection of lipopolysaccharide. We examined the differences in arthritis severities in the two genotypes. After intraperitoneal KS administration in phosphate-buffered saline (PBS) or PBS alone, we evaluated the potential of KS in ameliorating arthritis and protecting against cartilage damage in deficient mice. GAG release induced by IL-1alpha in the explants, and severity of arthritis were greater in GlcNAc6ST-1(-/-) mice than their WT littermates. Intraperitoneal KS administration effectively suppressed arthritis induction in GlcNAc6ST-1(-/-) mice. Thus, GlcNAc6ST-1(-/-) mice cartilage is more fragile than WT mice cartilage, and exogenous KS can suppress arthritis induction in GlcNAc6ST-1(-/-) mice. Vestigial KS chain or altered glycosylation in articular cartilage in GlcNAc6ST-1(-/-) mice may be protective against arthritis and associated cartilage damage as well as cartilage damage in culture. KS may offer therapeutic opportunities for chondroprotection and suppression of joint damage in inflammatory arthritis and may become a therapeutic agent for treating rheumatoid arthritis. PMID- 21624347 TI - Inhibition of selenocysteine tRNA[Ser]Sec aminoacylation provides evidence that aminoacylation is required for regulatory methylation of this tRNA. AB - There are two isoforms of selenocysteine (Sec) tRNA([Ser]Sec) that differ by a single methyl group, Um34. The non-Um34 isoform supports the synthesis of a subclass of selenoproteins, designated housekeeping, while the Um34 isoform supports the expression of another subclass, designated stress-related selenoproteins. Herein, we investigated the relationship between tRNA([Ser]Sec) aminoacylation and Um34 synthesis which is the last step in the maturation of this tRNA. Mutation of the discriminator base at position 73 in tRNA([Ser]Sec) dramatically reduced aminoacylation with serine, as did an inhibitor of seryl tRNA synthetase, SB-217452. Although both the mutation and the inhibitor prevented Um34 synthesis, neither precluded the synthesis of any other of the known base modifications on tRNA([Ser]Sec) following microinjection and incubation of the mutant tRNA([Ser]Sec) transcript, or the wild type transcript along with inhibitor, in Xenopus oocytes. The data demonstrate that Sec tRNA([Ser]Sec) must be aminoacylated for Um34 addition. The fact that selenium is required for Um34 methylation suggests that Sec must be attached to its tRNA for Um34 methylation. This would explain why selenium is essential for the function of Um34 methylase and provides further insights into the hierarchy of selenoprotein expression. PMID- 21624348 TI - MAPK/ERK and Wnt/beta-Catenin pathways are synergistically involved in proliferation of Sca-1 positive hepatic progenitor cells. AB - Hepatic progenitor cells (HPCs) persist in adulthood and have the potential to play a major role in regenerating diseased liver. However, the signaling pathways that both directly and indirectly regulate HPCs' self-renewal and differentiation remain elusive. Previously, we identified a bipotent, stem cell antigen-1 (Sca-1) positive HPC population from naive adult liver tissue. In the present study, we aimed to investigate the involvement of various signaling pathways in Sca-1(+) HPC proliferation. Epidermal growth factor (EGF) supplementation shows a significant increase in Sca-1(+) HPC proliferation and colony formation while stimulating phosphorylation of ERK1/2 and activating the induction of Cyclin D1. There were no demonstrable effects of EGF on Akt. The MEK inhibitor, PD0325901, inhibits proliferation and ERK1/2 phosphorylation while also suppressing the expression of Cyclin D1. In addition, activation of either IL-6/STAT3 or Wnt/beta Catenin pathway did not independently support cell proliferation and colony formation of HPCs. The Wnt/beta-Catenin pathway can cooperate with EGF to significantly promote HPC colony formation ratio and maintain long-term HPC in vitro. The data indicates that the MAPK/ERK pathway is both essential and critical for HPC proliferation, and the Wnt signaling pathway is not sufficient, while it works synergistically with the MAPK/ERK signaling pathway to promote HPC proliferation. PMID- 21624349 TI - Light responsiveness of clock genes, Per1 and Per2, in the olfactory bulb of mice. AB - The olfactory bulb (OB) of rodents has been suggested to possess a self sustaining circadian oscillator which functions independent from the master circadian clock in the suprachiasmatic nucleus (SCN) of the hypothalamus. However, neither histology nor physiology of this extra-SCN clock is studied yet. In the present study, we examined circadian variation of major clock gene expressions in the OB and responsiveness to single photic stimuli. Here we show significant circadian variation in the expression of clock genes, Per1, Per2 and Bmal1 in the OB. Per1 and PER2 were mainly expressed in the mitral cell and granular cell layers of the OB. Light responsiveness of Per1 and Per2 expression was different in the OB from that in the parietal cortex. Both Per1 and Per2 are expressed in the OB only by l000 lux light pulse, whereas 100 lux light was enough to induce Per1 mRNA in the parietal cortex. Interestingly, even 1000 lux light failed to induce Per2 mRNA in the parietal cortex. These clock gene specific and brain region-dependent responses to lights in the OB and parietal cortex suggest that single light stimulus induces various physiological functions in different brain areas via specific clock gene. PMID- 21624350 TI - ALCAPs induce mitochondrial apoptosis and activate DNA damage response by generating ROS and inhibiting topoisomerase I enzyme activity in K562 leukemia cell line. AB - Endemic Alkanna cappadocica was used to isolate novel antitumor molecules from Turkish landscapes in our previous studies. In this study, deoxyalkannin (ALCAP1), beta,beta-dimethylacrylalkannin (ALCAP2), acetylalkannin (ALCAP3), and alkannin (ALCAP4) as well as the novel isolated compounds 5-methoxydeoxyalkannin (ALCAP5), 8-methoxydeoxyalkannin (ALCAP6), 5-methoxyacetylalkannin (ALCAP7), 5 methoxy-beta,beta-dimethylacrylalkannin (ALCAP8) were characterized. The topoisomerase I (topo I) inhibitory activity of ALCAPs was investigated using in vitro plasmid relaxation assay and found that ALCAP2, 3, 4 and 7 were potent inhibitors at 2-6MUM concentrations. Further, DNA damage response to ALCAP treatments was also studied by measuring the H2AX((S139)) and ATM((S1981)) phosphorylations. ALCAP2, 7 and 8 induced the DNA damage and apoptosis, consistently resulted in PARP cleavage at nanomolar concentrations in K562 leukemia cells. Moreover, when the free radical (ROS) generating capacity of the compounds was studied by 2',7'-dichlorofluorescein-diacetate assay using flow cytometry, we found that a known antioxidant N-acetyl-cysteine almost completely abrogated the H2AX((S139)) phosphorylations and the caspase 3 cleavage and activation. Thus, gammaH2AX((S139)) foci formation remained higher than the control, and an increase in CHK2((T68)) phosphorylation was observed by ALCAP2 and 7 treatments suggested that, these compounds can be potential therapeutics against tumor cell growth because of their unique DNA damaging abilities additional to enzyme inhibition similar to those of doxorubicin. PMID- 21624351 TI - Fluoride enhances transfection activity of carbonate apatite by increasing cytoplasmic stability of plasmid DNA. AB - Intracellular delivery of a functional gene or a nucleic acid sequence to specifically knockdown a harmful gene is a potential approach to precisely treat a critical human disease. The intensive efforts in the last few decades led to the development of a number of viral and non-viral synthetic vectors. However, an ideal delivery tool in terms of the safety and efficacy has yet to be established. Recently, we have developed pH-sensing inorganic nanocrystals of carbonate apatite for efficient and cell-targeted delivery of gene and gene silencing RNA. Here we show that addition of very low level of fluoride to the particle-forming medium facilitates a robust increase in transgene expression following post-incubation of the particles with HeLa cells. Confocal microscopic observation and Southern blotting prove the cytoplasmic existence of plasmid DNA delivered by likely formed fluoridated carbonate apatite particles while degradation of plasmid DNA presumably by cytoplasmic nucleases was noticed following delivery with apatite particles alone. The beneficial role of fluoride in enhancing carbonate apatite-mediated gene expression might be due to the buffering potential of generated fluoridated apatite in endosomal acidic environment, thereby increasing the half-life of delivered plasmid DNA. PMID- 21624352 TI - NeuA O-acetylesterase activity is specific for CMP-activated O-acetyl sialic acid in Streptococcus suis serotype 2. AB - Several bacteria causing meningitis, such as Escherichia coli K1, Streptococcus suis, Neisseria meningitidis, and group B Streptococci (GBS), produce sialic acid (Neu5Ac)-containing capsular polysaccharide (CPS). Biosynthesis of the Neu5Ac containing CPS requires CMP-Neu5Ac as substrate, which is synthesized by CMP Neu5Ac synthetase from CTP and Neu5Ac. In E. coli or GBS, the NeuA protein encoded by the neuA gene has been known encoding a bifunctional enzyme that possesses both CMP-Neu5Ac synthetase and O-acetylesterase activity. In this report, we found that the S. suis NeuA (SsNeuA) was also a bifunctional CMP Neu5Ac synthetase/O-acetylesterase. Biochemical analyses revealed that the SsNeuA strictly de-O-acetylated CMP-O-acetyl-Neu5Ac, whereas the E. coli NeuA (EcNeuA) preferentially de-O-acetylated CMP-O-acetyl-Neu5Ac. E. coli devoid of NeuA O acetylesterase activity was unable to produce capsule and only CMP-Neu5Ac synthetase activity of the EcNeuA or SsNeuA could not restore its ability to produce capsule. These results suggest that the O-acetylesterase is essential for the synthesis of capsular Neu5Ac in E. coli, probably in S. suis and GBS as well. Our findings are key to understanding the biosynthesis of capsular Neu5Ac in E. coli, S. suis and GBS. PMID- 21624353 TI - Evidence of tricellulin expression by immune cells, particularly microglia. AB - Tight junctions (TJs) are elaborate structures located on the apical region of epithelial cells that limit paracellular permeability. Tricellulin is a recently discovered TJ protein, which is concentrated at the structurally specialized tricellular TJs but also present at bicellular contacts between epithelial cells, namely in the stomach. Interestingly, several TJ proteins have been found in other than epithelial cells, as astrocytes, and tricellulin mRNA expression was reported in mature dendritic cells. These findings prompted us to look for tricellulin expression in both epithelial and immune cells in the stomach, as well as in microglia, the brain resident immunocompetent cells. Immunohistochemical analysis of human stomach tissue sections revealed peroxidase staining at three-corner contact sites, as well as at the contact between two adjacent epithelial cells, thus evidencing the expression of tricellulin not only at tricellullar but at bicellular junctions as well. Such analysis, further revealed tricellulin immunostaining in cells of the monocyte/macrophage lineage, scattered throughout the lamina propria. Cultured rat microglia exhibited a notorious tricellulin staining, consistent with an extensive expression of the protein along the cell, which was not absolutely coincident with the lysosomal marker CD68. Detection of mRNA expression by real-time PCR provided supportive evidence for the expression of the TJ protein in microglia. These data demonstrate for the first time that microglia express a TJ protein. Moreover, the expression of tricellulin both in microglia and in the stomach immune cells point to a possible role of this new TJ protein in the immune system. PMID- 21624354 TI - The Akt/GSK-3beta pathway mediates flurbiprofen-induced neuroprotection against focal cerebral ischemia/reperfusion injury in rats. AB - Apoptosis is one of the major mechanisms of cell death during cerebral ischemia and reperfusion injury. Flurbiprofen has been shown to reduce cerebral ischemia/reperfusion injury in both focal and global cerebral ischemia models, but the mechanism remains unclear. This study aimed to investigate the potential association between the neuroprotective effect of flurbiprofen and the apoptosis inhibiting signaling pathways, in particularly the Akt/GSK-3beta pathway. A focal cerebral ischemia rat model was subjected to middle cerebral artery occlusion (MCAO) for 120 min and then treated with flurbiprofen at the onset of reperfusion. The infarct volume and the neurological deficit scores were evaluated at 24h after reperfusion. Cell apoptosis, apoptosis-related proteins and the levels of p-Akt and p-GSK-3beta in ischemic penumbra were measured using TUNEL and western blot. The results showed that administration of flurbiprofen at the doses of 5 and 10mg/kg significantly attenuated brain ischemia/reperfusion injury, as shown by a reduction in the infarct volume, neurological deficit scores and cell apoptosis. Moreover, flurbiprofen not only inhibited the expression of Bax protein and p-GSK-3beta, but also increased the expression of Bcl-2 protein, the ratio of Bcl-2/Bax as well as the P-Akt level. Taken together, these results suggest that flurbiprofen protects the brain from ischemia/reperfusion injury by reducing apoptosis and this neuroprotective effect may be partly due to the activation of Akt/GSK-3beta signaling pathway. PMID- 21624355 TI - The association of gamma-glutamyltransferase and C-reactive protein with IFG/IGT in Chinese adults in Qingdao, China. AB - BACKGROUND: Serum gamma-glutamyltransferase (GGT) and C-reactive protein(CRP) have been previously shown to be associated with impaired fasting glucose/impaired glucose tolerance (IFG/IGT), but such an association has not been well verified, and is examined in a non-diabetic Chinese population. METHODS: A population-based cross-sectional study was conducted in 2006 in Qingdao, China. Data of 1143 men and 1689 women aged 35-74 years and free of diabetes at baseline were analyzed. Multivariable logistic regression analysis was performed to estimate the odds ratio (OR) and its 95% confidence interval (CI). RESULTS: Compared with the lowest quartile, the ORs (95%CI) for IFG/IGT corresponding to the highest quartile were 0.89(0.61,1.28) in men and 0.87(0.64,1.18) in women for CRP and 2.12(1.40,3.38) and 1.87(1.32,2.62) for GGT, when the two were fitted simultaneously in a model adjusting for age, school years, alcohol-drinking, smoking, family history of diabetes, systolic blood pressure, waist circumference, triglycerides and high-density lipoprotein. CONCLUSIONS: The elevated GGT, but not CRP, was independently associated with the presence of the IFG/IGT in both genders in this Chinese population. PMID- 21624356 TI - Simple and precise detection of UGT1A1 polymorphisms with a modified loop-hybrid mobility shift assay using Cy5-labeled loop probes. AB - BACKGROUND: Irinotecan, an inhibitor of topoisomerase I, has been widely used as an important anti-cancer therapeutic drug. Deleterious effects of the drug in hypersensitive patients are known to be associated with genetic polymorphisms of the UGT1A1 gene, namely the polymorphic variants, *28 and *6. METHODS: A modified form of loop-hybrid mobility shift assay using a Cy5-tagged loop-hybrid probe was proposed as a precise and easy method of determining TA repeat polymorphisms at the *28 locus. RESULTS: In this modified method, only loop-hybrid bands were detected by a Cy5-fluorescent signal, despite several irregular electrophoretic bands due to TA repeats in the PCR product. CONCLUSIONS: When a loop-hybrid using a Cy5-tagged probe for the *28 locus and *6 locus were combined and used for mobility shift assay, simultaneous typing of the *28 and *6 variants was achieved in a single lane. PMID- 21624357 TI - Associations of atherosclerotic risk factors with oxidized low-density lipoprotein evaluated by LOX-1 ligand activity in healthy men. AB - BACKGROUND: The aim of this study was to determine the relationships of risk factors for atherosclerosis with oxidized low-density lipoprotein (OxLDL) evaluated by a new enzyme immunoassay for measurement of LOX-1 (lectin-like OxLDL receptor) ligand. METHODS: Subjects were 236 healthy men aged 33-62 years. LOX-1 ligand containing apoB (LAB) was measured by an enzyme-immunoassay using immobilized recombinant LOX-1 and anti-ApoB monoclonal antibody. RESULTS: In simple regression analysis, log-converted LAB showed significant positive correlations with history of smoking, waist circumference, diastolic blood pressure, LDL cholesterol, log-converted triglycerides, uric acid and white blood cell count and showed a significant negative correlation with HDL cholesterol. In multiple regression analysis using history of smoking, history of drinking, waist circumference, diastolic blood pressure, HDL cholesterol, log-converted triglycerides and uric acid as explanatory variables, log-converted LAB showed significant correlations only with history of smoking and log-converted triglycerides. Log-converted LAB was significantly higher in heavy smokers (>=20 cigarettes per day) than in nonsmokers and light smokers (<20 cigarettes per day), while no difference in log-converted LAB was found between nonsmokers and light smokers. Log-converted LAB was significantly higher in subjects with hypertriglyceridemia (>=150 mg/dl), large waist circumference (>=85 cm), high diastolic blood pressure (>=85 mmHg), or metabolic syndrome defined by the NCEP ATP III criteria than in subjects without each risk factor or metabolic syndrome. CONCLUSIONS: Hypertriglyceridemia and smoking are determinants of LOX-1 ligand activity in healthy men and are thus thought to be crucial risk factors for initiation of atherosclerotic progression through generation of OxLDL. PMID- 21624358 TI - Differentiation of the ductal epithelium and smooth muscle in the prostate gland are regulated by the Notch/PTEN-dependent mechanism. AB - We have shown previously that during branching morphogenesis of the mouse prostate gland, Bone morphogenetic protein 7 functions to restrict Notch1 positive progenitor cells to the tips of the prostate buds. Here, we employed prostate-specific murine bi-genic systems to investigate the effects of gain and loss of Notch function during prostate development. We show that Nkx3.1(Cre) and Probasin(Cre) alleles drive expression of Cre recombinase to the prostate epithelium and periepithelial stroma. We investigated the effects of gain of Notch function using the Rosa(NI1C) conditional allele, which carries a constitutively active intracellular domain of Notch1 receptor. We carried out the analysis of loss of Notch function in Nkx3.1(Cre/+);RBP-J(flox/flox) prostates, where RBP-J is a ubiquitous transcriptional mediator of Notch signaling. We found that gain of Notch function resulted in inhibition of the tumor suppressor PTEN, and increase in cell proliferation and progenitor cells in the basal epithelium and smooth muscle compartments. In turn, loss of Notch/RBP-J function resulted in decreased cell proliferation and loss of epithelial and smooth muscle progenitors. Gain of Notch function resulted in an early onset of benign prostate hyperplasia by three months of age. Loss of Notch function also resulted in abnormal differentiation of the prostate epithelium and stroma. In particular, loss of Notch signaling and increase in PTEN promoted a switch from myoblast to fibroblast lineage, and a loss of smooth muscle. In summary, we show that Notch signaling is necessary for terminal differentiation of the prostate epithelium and smooth muscle, and that during normal prostate development Notch/PTEN pathway functions to maintain patterned progenitors in the epithelial and smooth muscle compartments. In addition, we found that both positive and negative modulation of Notch signaling results in abnormal organization of the prostate tissue, and can contribute to prostate disease in the adult organ. PMID- 21624359 TI - The E3 ubiquitin ligase Cullin 4A regulates meiotic progression in mouse spermatogenesis. AB - The Cullin-RING ubiquitin-ligase CRL4 controls cell cycle and DNA damage checkpoint response and ensures genomic integrity. Inactivation of the Cul4 component of the CRL4 E3 ligase complex in Caenorhabditis elegans by RNA interference results in massive mitotic DNA re-replication in the blast cells, largely due to failed degradation of the DNA licensing protein, CDT-1, and premature spermatogenesis. Here we show that inactivation of Cul4a by gene targeting in mice only affected male but not female fertility. This male infertility phenotype resulted from a combination of decreased spermatozoa number, reduced sperm motility and defective acrosome formation. Agenesis of the mutant germ cells was accompanied by increased cell death in pachytene/diplotene cells with markedly elevated levels of phospho-p53 and CDT-1. Despite apparent normal assembly of synaptonemal complexes and DNA double strand break repair, dissociation of MLH1, a component of the late recombination nodule, was delayed in Cul4a -/- diplotene spermatocytes, which potentially led to subsequent disruptions in meiosis II and spermiogenesis. Together, our study revealed an indispensable role for Cul4a during male germ cell meiosis. PMID- 21624360 TI - Neurogenesis in the water flea Daphnia magna (Crustacea, Branchiopoda) suggests different mechanisms of neuroblast formation in insects and crustaceans. AB - Within euarthropods, the morphological and molecular mechanisms of early nervous system development have been analysed in insects and several representatives of chelicerates and myriapods, while data on crustaceans are fragmentary. Neural stem cells (neuroblasts) generate the nervous system in insects and in higher crustaceans (malacostracans); in the remaining euarthropod groups, the chelicerates (e.g. spiders) and myriapods (e.g. millipedes), neuroblasts are missing. In the latter taxa, groups of neural precursors segregate from the neuroectoderm and directly differentiate into neurons and glial cells. In all euarthropod groups, achaete-scute homologues are required for neuroblast/neural precursor group formation. In the insects Drosophila melanogaster and Tribolium castaneum achaete-scute homologues are initially expressed in clusters of cells (proneural clusters) in the neuroepithelium but expression becomes restricted to the future neuroblast. Subsequently genes such as snail and prospero are expressed in the neuroblasts which are required for asymmetric division and differentiation. In contrast to insects, malacostracan neuroblasts do not segregate into the embryo but remain in the outer neuroepithelium, similar to vertebrate neural stem cells. It has been suggested that neuroblasts are present in another crustacean group, the branchiopods, and that they also remain in the neuroepithelium. This raises the questions how the molecular mechanisms of neuroblast selection have been modified during crustacean and insect evolution and if the segregation or the maintenance of neuroblasts in the neuroepithelium represents the ancestral state. Here we take advantage of the recently published Daphnia pulex (branchiopod) genome and identify genes in Daphnia magna that are known to be required for the selection and asymmetric division of neuroblasts in the fruit fly D. melanogaster. We unambiguously identify neuroblasts in D. magna by molecular marker gene expression and division pattern. We show for the first time that branchiopod neuroblasts divide in the same pattern as insect and malacostracan neuroblasts. Furthermore, in contrast to D. melanogaster, neuroblasts are not selected from proneural clusters in the branchiopod. Snail rather than ASH is the first gene to be expressed in the nascent neuroblasts suggesting that ASH is not required for the selection of neuroblasts as in D. melanogaster. The prolonged expression of ASH in D. magna furthermore suggests that it is involved in the maintenance of the neuroblasts in the neuroepithelium. Based on these and additional data from various representatives of arthropods we conclude that the selection of neural precursors from proneural clusters as well as the segregation of neural precursors represents the ancestral state of neurogenesis in arthropods. We discuss that the derived characters of malacostracans and branchiopods - the absence of neuroblast segregation and proneural clusters - might be used to support or reject the possible groupings of paraphyletic crustaceans. PMID- 21624361 TI - Ceramide 1-phosphate induces neointimal formation via cell proliferation and cell cycle progression upstream of ERK1/2 in vascular smooth muscle cells. AB - Ceramide 1-phosphate (C1P) is a novel bioactive sphingolipid formed by ceramide kinase (CERK)-catalyzed phosphorylation of ceramide. It has been implicated in the regulation of such vital pathophysiological functions as phagocytosis and inflammation, but there have been no reports ascribing a biological function to CERK in vascular disorders. Here the potential role of CERK/C1P in neointimal formation was investigated using rat aortic vascular smooth muscle cells (VSMCs) in primary culture and a rat carotid injury model. Exogenous C8-C1P stimulated cell proliferation, DNA synthesis, and cell cycle progression of rat aortic VSMCs in primary culture. In addition, wild-type CERK-transfected rat aortic VSMCs induced a marked increase in rat aortic VSMC proliferation and [(3)H]-thymidine incorporation when compared to empty vector transfectant. C8-C1P markedly activated extracellular signal-regulated kinase 1 and 2 (ERK1/2) within 5min, and the activation could be prevented by U0126, a MEK inhibitor. Also, K1, a CERK inhibitor, decreased the ERK1/2 phosphorylation and cell proliferation on platelet-derived growth factor (PDGF)-stimulated rat aortic VSMCs. CERK expression and C1P levels were found to be potently increased during neointimal formation using a rat carotid injury model. However, ceramide levels decreased during the neointimal formation process. These findings suggest that C1P can induce neointimal formation via cell proliferation through the regulation of the ERK1/2 protein in rat aortic VSMCs and that CERK/C1P may regulate VSMC proliferation as an important pathogenic marker in the development of cardiovascular disorders. PMID- 21624362 TI - ZNF689 suppresses apoptosis of hepatocellular carcinoma cells through the down regulation of Bcl-2 family members. AB - ZNF689, a C2H2-type of zinc finger transcription factor, was suggested to play a key role in hepatocarcinogenesis. However, none of the target genes or potential roles of ZNF689 in hepatocellular carcinoma (HCC) have been elucidated. Here, we investigated the role of ZNF689 in HCC cell lines focusing on cell viability and apoptosis.We found that the knockdown of ZNF689 by its specific siRNA decreased cell viability of Huh7. Cell cycle analysis revealed that the ZNF689 knockdown increased the proportion of the sub-G1 population, accompanied by an increase of annexin V- and TUNEL-positive cells.Western blot analysis revealed that ZNF689 knockdown induced the expression of pro-apoptotic factors of Bcl-2 family, Bax, Bak and jBid. There was a correlation between the expression of ZNF689 and an anticancer drug 5-fluorouracil (5-FU) resistance of HCC cells. In vivo, ZNF689 siRNA reduced tumor viability in HepG2-bearing mice with statistical significance. Furthermore, immunohistochemical analysis demonstrated that nuclei of a significant portion of human HCC surgical specimens were positive for ZNF689. Taken together, our results indicate that ZNF689 blocks pro-apoptotic signaling by suppressing the Bak/Bax/Bid pathway, resulting in the progression of liver cancer and resistance to 5-FU. ZNF689 may be a promising chemotherapeutic target against liver cancer. PMID- 21624363 TI - Heregulin activation of ErbB2/ErbB3 signaling potentiates the integrity of airway epithelial barrier. AB - BACKGROUND: Members of the ErbB family of the receptor protein tyrosine kinase superfamily mediate heregulin (HRG)-induced cell responses. Here we investigated HRG activation of ErbB receptors, and the role of this activation in the development of the permeability barrier in airway epithelial cells (AECs). METHODS: Two airway epithelial-like cell lines, Calu-3 and 16HBE were exposed to HRG or no stimulus and were evaluated with respect to their paracellular permeability as determined by transepithelial electric resistance (TER) and fluorescein isothiocyanate (FITC)-dextran flux. Tight junctions (TJs) were assessed by immunocytochemical localization of occludin and zonula occludens-1. RESULTS: HRG promoted the development of the permeability barrier and TJ formation by monolayers of Calu-3 and 16HBE cells. Calu-3 cells expressed ErbB1, ErbB2, and ErbB3, but not ErbB4, on their surface. ErbB3 knockdown by small interference RNA (siRNA) blunted the effects of HRG on the permeability barrier. ErbB3 is known as a kinase-dead receptor and relies on other members of the family for its phosphorylation. To identify its heterodimerization partner, we knocked down the expression of other ErbB family receptors. We found that HRG's effect on the permeability barrier could be significantly attenuated by transfecting cells with ErbB2 siRNA but not with EGFR siRNA. CONCLUSION: These results indicate that HRG activation of ErbB2/ErbB3 heterodimers is essential for regulation of the permeability barrier in AECs. PMID- 21624364 TI - Cimetidine enhances the protective effect of GST DNA vaccine against Schistosoma japonicum. AB - Cimetidine (CIM), a histamine-2-receptor antagonist, has a long history of safe use in gastric acid-mediated gastrointestinal disorders. In this study, we used CIM, as an adjuvant, with pEGFP-Sj26 GST (the recombinant plasmid containing enhanced green fluorescent protein gene and the gene encoding 26 kDa glutathione S-transferase of Schistosoma japonicum) DNA vaccine to immunized mice and attempted to enhance the protective effect against S. japonicum. The results showed that the reduction rate of worm and egg burdens in the pEGFP-Sj26GST plus CIM group were 79.0% and 68.4%, respectively, significantly higher than that in pEGFP-Sj26GST alone group (27.0% and 22.5%, P<0.01). Compared with the pEGFP Sj26GST alone group, mice immunized with pEGFP-Sj26GST plus CIM showed an elevated level of IFN-gamma and IL-12 and a low level of IL-10 in splenocytes, while the levels of IL-4 and IL-5 showed no difference between the two groups. Our data also demonstrated that the percentage of CD4(+)CD25(+) regulatory T cells (Tregs) was significantly decreased in the spleens of mice immunized with pEGFP-Sj26GST plus CIM. All these findings suggest that CIM as a potential schistosome DNA vaccine adjuvant can enhance the protective effect of pEGFP Sj26GST vaccine. PMID- 21624365 TI - Allosteric nucleotide-binding site in the mitochondrial NADH:ubiquinone oxidoreductase (respiratory complex I). AB - The rotenone-insensitive NADH:hexaammineruthenium III (HAR) oxidoreductase reactions catalyzed by bovine heart and Yarrowia lipolytica submitochondrial particles or purified bovine complex I are stimulated by ATP and other purine nucleotides. The soluble fraction of mammalian complex I (FP) and prokaryotic complex I homolog NDH-1 in Paracoccus denitrificans plasma membrane lack stimulation of their activities by ATP. The stimulation appears as a decrease in apparent K(m) values for NADH and HAR. Thus, the "accessory" subunits of eukaryotic complex I bear an allosteric ATP-binding site. PMID- 21624366 TI - Innate immune responses in central nervous system inflammation. AB - In autoimmune diseases of the central nervous system (CNS), innate glial cell responses play a key role in determining the outcome of leukocyte infiltration. Access of leukocytes is controlled via complex interactions with glial components of the blood-brain barrier that include angiotensin II receptors on astrocytes and immunoregulatory mediators such as Type I interferons which regulate cellular traffic. Myeloid cells at the blood-brain barrier present antigen to T cells and influence cytokine effector function. Myelin-specific T cells interact with microglia and promote differentiation of oligodendrocyte precursor cells in response to axonal injury. These innate responses offer potential targets for immunomodulatory therapy. PMID- 21624367 TI - p53 regulation by ubiquitin. AB - The ubiquitination pathway is a highly dynamic and coordinated process that regulates degradation as well as numerous processes of proteins within a cell. The p53 tumor suppressor and several factors in the pathway are regulated by ubiquitin as well as ubiquitin-like proteins. These modifications are critical for the function of p53 and control both the degradation of the protein as well as localization and activity. Importantly, more recent studies have identified deubiquitination enzymes that can specifically remove ubiquitin moieties from p53 or other factors in the pathway, and the reversible nature of this process adds yet another layer of regulatory control of p53. This review highlights the recent advances in our knowledge of ubiquitin and the p53 pathway. PMID- 21624368 TI - Locust phase polyphenism: Does epigenetic precede endocrine regulation? AB - The morphological, physiological and behavioural differences between solitarious and gregarious desert locusts are so pronounced that one could easily mistake the two phases as belonging to different species, if one has no knowledge of the phenomenon of phenotypic plasticity. A number of phase-specific features are hormonally controlled. Juvenile hormone promotes several solitarious features, the green cuticular colour being the most obvious one. The neuropeptide corazonin elicits the dark cuticular colour that is typical for the gregarious phase, as well as particular gregarious behavioural characteristics. However, it had to be concluded, for multiple reasons, that the endocrine system is not the primary phase-determining system. Our observation that longevity gets imprinted in very early life by crowding of the young hatchlings, and that it cannot be changed thereafter, made us consider the possibility that, perhaps, epigenetic control of gene expression might be, if not the missing, a primary phase-determining mechanism. Imprinting is likely to involve DNA methylation and histone modification. Analysis of a Schistocerca EST database of nervous tissue identified the presence of several candidate genes that may be involved in epigenetic control, including two DNA methyltransferases (Dnmts). Dnmt1 and Dnmt2 are phase-specifically expressed in certain tissues. In the metathoracic ganglion, important in the serotonin pathway for sensing mechanostimulation, their expression is clearly affected by crowding. Our data urge for reconsidering the role of the endocrine system as being sandwiched in between genetics and epigenetics, involving complementary modes of action. PMID- 21624369 TI - Salinity-dependent in vitro effects of homologous natriuretic peptides on the pituitary-interrenal axis in eels. AB - We examined the effects of atrial, B-type, ventricular and C-type natriuretic peptides (ANP, BNP, VNP and CNP1, 3, 4) on cortisol secretion from interrenal tissue in vitro in both freshwater (FW) and seawater (SW)-acclimated eels. We first localized the interrenal and chromaffin cells in the eel head kidney using cell specific markers (cholesterol side-chain cleavage enzyme (P450ssc) and tyrosine hydroxylase (TH), respectively) and established the in vitro incubation system for eel interrenal tissue. Unexpectedly, none of the NPs given alone to the interrenal tissue of FW and SW eels stimulated cortisol secretion. However, ANP and VNP, but not BNP and three CNPs, enhanced the steroidogenic action of ACTH in SW interrenal preparations, while CNP1 and CNP4, but not ANP, BNP, VNP and CNP3, potentiated the ACTH action in FW preparations. These salinity dependent effects of NPs are consistent with the previous in vivo study in the eel where endogenous ACTH can act with the injected NPs. 8-Br-cGMP also enhanced the ACTH action in both FW and SW eel preparations, suggesting that the NP actions were mediated by the guanylyl cyclase-coupled NP receptors (GC-A and B) that were localized in the eel interrenal. Further, ANP and CNP1 stimulated ACTH secretion from isolated pituitary glands of SW and/or FW eels. In summary, the present study revealed complex mechanisms of NP action on corticosteroidogenesis through the pituitary-interrenal axis in eels, thereby providing a deeper insight into the role of the NP family in the acclimation of this euryhaline teleost to diverse salinity environments. PMID- 21624370 TI - Environmental influences on Adelie penguin breeding schedules, endocrinology, and chick survival. AB - To understand how the social and physical environment influences behaviour, reproduction and survival, studies of underlying hormonal processes are crucial; in particular, interactions between stress and reproductive responses may have critical influences on breeding schedules. Several authors have examined the timing of breeding in relation to environmental stimuli, while others have independently described endocrine profiles. However, few studies have simultaneously measured endocrine profiles, breeding behaviour, and offspring survival across seasons. We measured sex and stress hormone concentrations (oestrogens, testosterone, and corticosterone), timing of breeding, and chick survival, in Adelie penguins (Pygoscelis adeliae) at two colonies in two different years. Clutch initiation at Cape Bird South (CBS; year 1, ~14,000 pairs) occurred later than at Cape Crozier East (CCE; year 2, ~ 25,000 pairs); however, breeding was more synchronous at CBS. This pattern was probably generated by the persistence of extensive sea ice at CBS (year 1). Higher corticosterone metabolite and lower sex hormone concentrations at CBS correlated with later breeding and lower chick survival compared to at CCE - again, a likely consequence of sea ice conditions. Within colonies, sub-colony size (S, 50-100; M, 200-300; L, 500-600; XL, >1000 pairs) did not influence the onset or synchrony of breeding, chick survival, or hormone concentrations. We showed that the endocrine profiles of breeding Adelie penguins can differ markedly between years and/or colonies, and that combining measures of endocrinology, behaviour, and offspring survival can reveal the mechanisms and consequences that different environmental conditions can have on breeding ecology. PMID- 21624371 TI - The effects of experimentally infecting Australian tree frogs with lungworms (Rhabdias pseudosphaerocephala) from invasive cane toads. AB - Invasive species may transmit novel pathogens to native taxa, and lacking a history of coevolutionary interactions with the pathogen, the new hosts may be severely affected. Cane toads (Rhinella marina) were introduced to Australia in 1935, bringing with them a lungworm (Rhabdias pseudosphaerocephala) not found in Australian frogs. Previous studies suggest that most frog species are unaffected by this parasite, but one tree-frog (Litoria caerulea) can harbour high numbers of lungworm. More detailed laboratory studies confirm and extend the earlier results on L. caerulea and show that Rhabdias infection severely depresses the viability of metamorphs of an allied tree-frog species, Litoria splendida. Parasitic larvae infected both of these two closely related tree-frog species, but the two anurans differed in the consequences of infection. Parasitism reduced the survivorship of L. splendida and the stamina of both species. Lungworms did not consistently reduce growth rates or affect heart rates in either tree-frog species. Although L. splendida is potentially vulnerable to the arrival of toad transported lungworms, rates of host-switching may be reduced by low levels of habitat overlap between the frogs (which are rock-dwelling and arboreal) and the toads (which are terrestrial and most abundant in disturbed habitats). PMID- 21624372 TI - Absence of thrombospondin-2 increases cardiomyocyte damage and matrix disruption in doxorubicin-induced cardiomyopathy. AB - Clinical use of the antineoplastic agent doxorubicin (DOX) is limited by its cardiomyocyte toxicity. Attempts to decrease cardiomyocyte injury showed promising results in vitro, but failed to reduce the adverse effects of DOX in vivo, suggesting that other mechanisms contribute to its cardiotoxicity as well. Evidence that DOX also induces cardiac injury by compromising extracellular matrix integrity is lacking. The matricellular protein thrombospondin-2 (TSP-2) is known for its matrix-preserving function, and for modulating cellular function. Here, we investigated whether TSP-2 modulates the process of doxorubicin-induced cardiomyopathy (DOX-CMP). TSP-2-knockout (TSP-2-KO) and wild type (WT) mice were treated with DOX (2 mg/kg/week) for 12 weeks to induce DOX CMP. Mortality was significantly increased in TSP-2-KO compared to WT mice. Surviving DOX-treated TSP-2-KO mice had depressed cardiac function compared to WT animals, accompanied by increased cardiomyocyte apoptosis and matrix damage. Enhanced myocyte damage in the absence of TSP-2 was associated with impaired activation of the Akt signaling pathway in TSP-2-KO compared to WT. The absence of TSP-2, in vivo and in vitro, reduced Akt activation both under non-treated conditions and after DOX. Importantly, inhibition of Akt phosphorylation in cardiomyocytes significantly reduced TSP-2 expression, unveiling a unique feedback loop between Akt and TSP-2. Finally, enhanced matrix disruption in DOX treated TSP-2-KO hearts went along with increased matrix metalloproteinase-2 levels. Taken together, this study is the first to provide evidence for the implication of the matrix element TSP-2 in protecting against DOX-induced cardiac injury and dysfunction. PMID- 21624373 TI - Three independent mechanisms contribute to tetracaine inhibition of cardiac calcium release channels. AB - Tetracaine is a tertiary amine local anaesthetic which inhibits ryanodine receptors (RyRs), the calcium release channels of the sarcoplasmic reticulum (SR). Tetracaine has been extensively used to study the role of the SR Ca(2+) fluxes in muscle cells, yet a detailed understanding of tetracaine action on RyR channels is lacking. Here we investigate tetracaine effects in single channel recording of sheep cardiac RyRs in lipid bilayers. Tetracaine decreased channel conductance (block) and open probability (inhibition). The IC(50) for inhibition had complex dependencies on membrane voltage and cytoplasmic [ATP], [Ca(2+)] and pH. We identify three mechanisms underlying these actions. First, a voltage dependent, slow inhibition in which luminal and cytoplasmic tetracaine compete for a common neutral/cation binding site within the trans-membrane RyR domain to induce long closed events (~100 ms). The apparent binding rate is proportional to the RyR closed probability, indicating that it only operates on closed channels. Second, a voltage-independent, pH sensitive fast inhibition in which cytoplasmic and luminal tetracaine compete for a site located on the cytoplasmic domain of the RyR to induce fast closed events (~2 ms). Its IC(50) is not dependent on the open/closed conformation of RyR. Finally, a voltage-dependent block of the channel by cytoplasmic tetracaine reduced channel conductance. We develop a model for tetracaine inhibition which predicts that under diastolic conditions, i.e. when RyRs are mainly closed, the slow mechanism has the highest potency (IC(50)~200 MUM) of the three mechanisms and is therefore the dominant form of inhibition. However, during periods of Ca(2+) release, i.e. when RyRs are open, the slow mechanism becomes ineffective, leaving the fast inhibition (IC(50)~2 mM) as the dominant effect. Because of this closed state inhibition property, tetracaine loses its efficacy when RyRs open. This has the effect of increasing the feedback on SR Ca(2+) release generated by cytoplasmic and luminal Ca(2+). PMID- 21624374 TI - Rapid pyrophosphate release from transcriptional elongation complexes appears to be coupled to a nucleotide-induced conformational change in E. coli core polymerase. AB - In the nucleotide addition cycle, pyrophosphate is generated upon incorporation of each nucleotide. Rapid release of pyrophosphate is essential for facile transcription elongation. Stopped-flow kinetic studies involving alterations in the intrinsic protein fluorescence of the core polymerase upon the binding of pyrophosphate to well-defined elongation complexes (ECs) indicate that the intrinsic off-rate of pyrophosphate (k=5.7-8.1 s(-1)) is too slow to account for the rapid rate of nucleotide incorporation that occurs during processive transcription elongation. Stopped-flow kinetic studies on UTP binding followed by UMP incorporation into an EC as monitored by alterations in the intrinsic protein fluorescence of the core polymerase resulted in a set of first-order rate constants that varied in a hyperbolic manner as a function of UTP concentration. This is consistent with a binding step (K(UTP)=17+/-6 MUM) followed by a conformational change (k=623+/-54 s(-1)) in the core polymerase. In comparable studies on ATP binding and AMP incorporation into an EC, the data were also consistent with a binding step (K(ATP)=44+/-6 MUM) followed by a conformational change (k=411+/-51 s(-1)) in the core polymerase. In stopped-flow kinetic studies with alpha,beta-methyleneadenosine 5' triphosphate, which can bind to the EC but cannot lead to nucleotide incorporation, the analysis of the hyperbolic dependence of the observed first-order rate constant on alpha,beta methyleneadenosine 5' triphosphate concentration yielded a value of 20+/-13 MUM for the apparent dissociation constant and a value of 221+/-36 s(-1) for the first-order rate constant for the associated conformational change in the core polymerase. This indicates that the conformational change in the core polymerase precedes chemistry. In conjunction with previously reported results on the increase in the rate of pyrophosphate release in the presence of the next cognate nucleotide for incorporation, the data are consistent with a model in which rapid pyrophosphate release is coupled to a conformational change in the core polymerase that precedes chemistry and that occurs upon the binding of the next cognate nucleotide for incorporation. PMID- 21624375 TI - Structure of the BamC two-domain protein obtained by Rosetta with a limited NMR data set. AB - The CS-RDC-NOE Rosetta program was used to generate the solution structure of a 27-kDa fragment of the Escherichia coli BamC protein from a limited set of NMR data. The BamC protein is a component of the essential five-protein beta-barrel assembly machine in E. coli. The first 100 residues in BamC were disordered in solution. The Rosetta calculations showed that BamC101-344 forms two well-defined domains connected by an ~18-residue linker, where the relative orientation of the domains was not defined. Both domains adopt a helix-grip fold previously observed in the Bet v 1 superfamily. 15N relaxation data indicated a high degree of conformational flexibility for the linker connecting the N-terminal domain and the C-terminal domain in BamC. The results here show that CS-RDC-NOE Rosetta is robust and has a high tolerance for misassigned nuclear Overhauser effect restraints, greatly simplifying NMR structure determinations. PMID- 21624376 TI - A generalization of Hamilton's rule--love others how much? AB - According to Hamilton's (1964a, b) rule, a costly action will be undertaken if its fitness cost to the actor falls short of the discounted benefit to the recipient, where the discount factor is Wright's index of relatedness between the two. We propose a generalization of this rule, and show that if evolution operates at the level of behavior rules, rather than directly at the level of actions, evolution will select behavior rules that induce a degree of cooperation that may differ from that predicted by Hamilton's rule as applied to actions. In social dilemmas there will be less (more) cooperation than under Hamilton's rule if the actions are strategic substitutes (complements). Our approach is based on natural selection, defined in terms of personal (direct) fitness, and applies to a wide range of pairwise interactions. PMID- 21624377 TI - A sensitivity study of conductivity values in the passive bidomain equation. AB - There is a complex interplay between the four conductivity values used in the bidomain equation and the resulting electric potential distribution in cardiac tissue arising from subendocardial ischaemia. Based on the three commonly used experimentally derived conductivity data sets, a non-dimensional formulation of the passive bidomain equation is derived, which gives rise naturally to several dimensionless conductivity ratios. The data sets are then used to define a parameter space of these ratios, which is studied by considering the correlation coefficients between different epicardial potential distributions. From this study, it is shown that the ratio of the intracellular longitudinal conductivity to the intracellular transverse conductivity is the key parameter in explaining the differences between the epicardial potential distributions observed with these three data sets. PMID- 21624378 TI - A dual negative regulation model of Toll-like receptor 4 signaling for endotoxin preconditioning in human endotoxemia. AB - We discuss a model illustrating how the outcome of repeated endotoxin administration experiments can emerge as a natural consequence of the tightly regulated signaling pathways and also highlight the importance of a dual negative feedback regulation including PI3K/Akt and IRAK-M (IRAK3). We identify the relative time scales of the onset and the magnitude of the stimulus as key determinants of outcome in repeated administration experiments. The results of our simulations involve potentiated response, tolerance, and protective tolerance. Moreover, the knockout of negative regulators shows that IRAK-M is a necessary and sufficient factor for generation of endotoxin tolerance (ET). The effects of the knockout of IRAK-M gene or administration of PI3K inhibitor do yield predictions that have been verified experimentally. Finally, the pretreatment with PI3K inhibitor reveals the interaction between these two negative regulations. PMID- 21624379 TI - Robust synchronization analysis in nonlinear stochastic cellular networks with time-varying delays, intracellular perturbations and intercellular noise. AB - Naturally, a cellular network consisted of a large amount of interacting cells is complex. These cells have to be synchronized in order to emerge their phenomena for some biological purposes. However, the inherently stochastic intra and intercellular interactions are noisy and delayed from biochemical processes. In this study, a robust synchronization scheme is proposed for a nonlinear stochastic time-delay coupled cellular network (TdCCN) in spite of the time varying process delay and intracellular parameter perturbations. Furthermore, a nonlinear stochastic noise filtering ability is also investigated for this synchronized TdCCN against stochastic intercellular and environmental disturbances. Since it is very difficult to solve a robust synchronization problem with the Hamilton-Jacobi inequality (HJI) matrix, a linear matrix inequality (LMI) is employed to solve this problem via the help of a global linearization method. Through this robust synchronization analysis, we can gain a more systemic insight into not only the robust synchronizability but also the noise filtering ability of TdCCN under time-varying process delays, intracellular perturbations and intercellular disturbances. The measures of robustness and noise filtering ability of a synchronized TdCCN have potential application to the designs of neuron transmitters, on-time mass production of biochemical molecules, and synthetic biology. Finally, a benchmark of robust synchronization design in Escherichia coli repressilators is given to confirm the effectiveness of the proposed methods. PMID- 21624381 TI - An introduction to the special issue on the neuropsychology of prospective memory. PMID- 21624380 TI - Focal adhesion kinase and endothelial cell apoptosis. AB - Focal adhesion kinase (FAK) is a key component of cell-substratum adhesions, known as focal adhesion complexes. Growing evidence indicates that FAK is important in maintenance of normal cell survival and that disruption of FAK signaling results in loss of substrate adhesion and anoikis (apoptosis) of anchorage-dependent cells, such as endothelial cells. Basal FAK activity in non stimulated endothelial cells is important in maintaining cell adhesion to integrins via PI3 kinase/Akt signaling. FAK activity is dependent upon small GTPase signaling. FAK also appears to be important in cardiomyocyte hypertrophy and hypoxia/reoxygenation-induced cell death. This review summarizes the signaling pathways of FAK in prevention of apoptosis and the role of FAK in mediating adenosine and homocysteine-induced endothelial cell apoptosis and in cardiovascular diseases. PMID- 21624382 TI - Multi-walled carbon nanotube-induced gene expression in the mouse lung: association with lung pathology. AB - Due to the fibrous shape and durability of multi-walled carbon nanotubes (MWCNT), concerns regarding their potential for producing environmental and human health risks, including carcinogenesis, have been raised. This study sought to investigate how previously identified lung cancer prognostic biomarkers and the related cancer signaling pathways are affected in the mouse lung following pharyngeal aspiration of well-dispersed MWCNT. A total of 63 identified lung cancer prognostic biomarker genes and major signaling biomarker genes were analyzed in mouse lungs (n=80) exposed to 0, 10, 20, 40, or 80MUg of MWCNT by pharyngeal aspiration at 7 and 56days post-exposure using quantitative PCR assays. At 7 and 56days post-exposure, a set of 7 genes and a set of 11 genes, respectively, showed differential expression in the lungs of mice exposed to MWCNT vs. the control group. Additionally, these significant genes could separate the control group from the treated group over the time series in a hierarchical gene clustering analysis. Furthermore, 4 genes from these two sets of significant genes, coiled-coil domain containing-99 (Ccdc99), muscle segment homeobox gene-2 (Msx2), nitric oxide synthase-2 (Nos2), and wingless-type inhibitory factor-1 (Wif1), showed significant mRNA expression perturbations at both time points. It was also found that the expression changes of these 4 overlapping genes at 7days post-exposure were attenuated at 56days post-exposure. Ingenuity Pathway Analysis (IPA) found that several carcinogenic-related signaling pathways and carcinogenesis itself were associated with both the 7 and 11 gene signatures. Taken together, this study identifies that MWCNT exposure affects a subset of lung cancer biomarkers in mouse lungs. PMID- 21624383 TI - Attention to faces: effects of face inversion. AB - Attention may be biased towards faces but a face advantage may be linked to the upright orientation of a face. Three experiments, employing a flanker and a cuing paradigm, investigated effects of face orientation, perceptual load and allocation of attention. Experiment 1 demonstrated that, irrespective of load, attention is biased towards upright face distractors while inverted face distractors are easy to ignore. Experiment 2 verified that inverted face distractors can interfere provided that they are attended to volitionally, likely because the volitional allocation of attention promotes face processing and gender classification (Experiment 3). PMID- 21624384 TI - Dopamine D1/5 and D2/3 agonists differentially attenuate somatic signs of nicotine withdrawal in rats. AB - Abrupt tobacco/nicotine cessation after chronic use causes various withdrawal symptoms/signs. There is evidence that dysfunction of brain dopaminergic system might be responsible for some nicotine withdrawal symptoms. The hypothesis for the present study was that different dopaminergic agonists would relieve different nicotine withdrawal signs. Adult male Sprague-Dawley rats were used. ( )-Nicotine bitartrate (9 mg/kg/day, salt content) or equimolar sodium tartrate was infused into each rat via a subcutaneous (s.c.) osmotic minipump for 7 days. To assess nicotine withdrawal signs, several somatic abstinence signs including teeth-chattering/chews, stretches/gasps, ptosis, shakes, and yawns were counted one day after removal of pumps. These signs were attenuated by the s.c. injection of 0.4 mg/kg nicotine bitartrate. Both a dopamine D(1/5) agonist (SKF81297) and a D(2/3) agonist (pramipexole) relieved abstinence signs dose-dependently but differentially. SKF81297 (0.32 mg/kg, s.c.) reduced teeth-chattering/chews but not shakes. Pramipexole (1mg/kg, s.c.) decreased both teeth-chattering/chews and shakes. A low dose of pramipexole (0.1mg/kg, s.c.) significantly increased yawns, consistent with previous studies that the stimulation of D(3) receptors induces yawning. These results indicate that a D(2)-selective agonist should be considered a candidate to relieve nicotine withdrawal symptoms. PMID- 21624385 TI - Concentration- and time-dependent behavioral changes in Caenorhabditis elegans after exposure to nicotine. AB - Nicotine induces profound behavioral responses in the model organism Caenorhabditis elegans. We tested the effect of a broad range of concentrations of nicotine (from 0.001 mM to 30 mM in nematode growth medium) on C. elegans locomotor behavior. We also followed the time-course influence on the sensitivity of C. elegans to nicotine (from 0 min to 300 min). A low concentration (0.001 mM) of this alkaloid causes a reduction of the speed of movement. By contrast, moderate concentrations (0.01 and 0.1 mM) induced acceleration of the mean speed of locomotion of C. elegans. High doses of nicotine (above 1 mM) induced slowing down of the movements and, finally, paralysis. Time-dependent analysis revealed that the stimulating effect of nicotine abolished the slowing down of C. elegans in control experiments after 30 min in the presence of 0.001, 0.1 and 10 mM nicotine. In the presence of 0.1 mM nicotine, the stimulation phase lasted up to 70 min. The evidence indicates that nicotine can have dual effects on the speed of locomotion, which is dependent on differences in its dosage and treatment time. PMID- 21624386 TI - Beneficial effects of resveratrol on scopolamine but not mecamylamine induced memory impairment in the passive avoidance and Morris water maze tests in rats. AB - Resveratrol (3,5,4-trihydroxy-trans-stilbene), which is found in grapes and red wine has been shown to protect neuronal cells with its antioxidant activity, improve memory function in dementia and reverse acetylcholine esterase (AChE) activity. The aim of this study was to investigate the effect of resveratrol on emotional and spatial memory in naive rats, as well as on scopolamine- and mecamylamine-induced memory impairment in the passive avoidance and Morris water maze (MWM) tests. Resveratrol (12.5, 25 and 50 mg/kg), scopolamine (0.6 mg/kg) and mecamylamine (10mg/kg) were administered to male Wistar rats. In the passive avoidance test, there was no significant difference in the first day latency between all groups, whereas scopolamine and mecamylamine significantly shortened the second day latency compared to the control group. Resveratrol reversed the effect of scopolamine at all doses used, but it had no effect on mecamylamine induced memory impairment in the passive avoidance test. Both scopolamine and mecamylamine significantly decreased the time spent in the escape platform quadrant during the probe trial of the MWM test compared to the control group. Resveratrol reversed the effect of scopolamine at all doses, but did not change the effect of mecamylamine in the MWM test. There were no significant differences in the locomotor activities of any of the groups. In conclusion, we suggested that resveratrol had improving effects on learning and memory by acting on muscarinic cholinergic receptors and at least in part, may reverse AChE activity. PMID- 21624387 TI - Stimulus control by 5-methoxy-N,N-dimethyltryptamine in wild-type and CYP2D6 humanized mice. AB - In previous studies we have observed that, in comparison with wild type mice, Tg CYP2D6 mice have increased serum levels of bufotenine [5-hydroxy-N,N dimethyltryptamine] following the administration of 5-MeO-DMT. Furthermore, following the injection of 5-MeO-DMT, harmaline was observed to increase serum levels of bufotenine and 5-MeO-DMT in both wild-type and Tg-CYP2D6 mice. In the present investigation, 5-MeO-DMT-induced stimulus control was established in wild type and Tg-CYP2D6 mice. The two groups did not differ in their rate of acquisition of stimulus control. When tested with bufotenine, no 5-MeO-DMT appropriate responding was observed. In contrast, the more lipid soluble analog of bufotenine, acetylbufotenine, was followed by an intermediate level of responding. The combination of harmaline with 5-MeO-DMT yielded a statistically significant increase in 5-MeO-DMT-appropriate responding in Tg-CYP2D6 mice; a comparable increase occurred in wild-type mice. In addition, it was noted that harmaline alone was followed by a significant degree of 5-MeO-DMT-appropriate responding in Tg-CYP2D6 mice. It is concluded that wild-type and Tg-CYPD2D6 mice do not differ in terms of acquisition of stimulus control by 5-MeO-DMT or in their response to bufotenine and acetylbufotenine. In both groups of mice, harmaline was found to enhance the stimulus effects of 5-MeO-DMT. PMID- 21624388 TI - Guamanian Suncus murinus responsiveness to emetic stimuli and the antiemetic effects of 8-OH-DPAT. AB - The Japanese Suncus murinus, the house musk shrew, is a small insectivore commonly used in emetic research. The Guamanian S. murinus has not had extensive testing as an emetic model, but it is readily available for use in emetic experiments in the United States, unlike the Japanese Suncus. This study determined that Guamanian S. murinus is an acceptable model for emesis research and its differences from the Japanese strain were examined. Motion and nicotine were used as emetic stimuli and comparable doses of 8-OH-DPAT were used to compare emetic susceptibility to the Japanese strain. The Guamanian strain had decreased susceptibility to motion and increased susceptibility to nicotine as compared to the Japanese, as well as increased sensitivity to 8-OH-DPAT, with lower doses of the recovery drug eliminating retching episodes. The study also determined that Guamanian S. murinus are smaller and more aggressive than the Japanese strain, but just as effective as a model for emetic research. PMID- 21624389 TI - Hesperidin induces antinociceptive effect in mice and its aglycone, hesperetin, binds to MU-opioid receptor and inhibits GIRK1/2 currents. AB - This paper extended the evaluation of the depressant and antinociceptive activities of hesperidin in order to determine its effectiveness by the intraperitoneal and oral routes, its pharmacological interaction with diverse pathways of neurotransmission and the role of its aglycone, hesperetin. The capacity of hesperidin and hesperetin to bind to MU-opioid receptor and their actions on MU-opioid receptor co-expressed with GIRK1/GIRK2 channels (G protein activated inwardly rectifying K+ channels) in Xenopus laevis oocytes were also determined. Hesperidin exhibited a depressant activity in the hole board and locomotor activity tests, antinociceptive activities in the abdominal writhing and hot plate tests and no motor incoordination in the inverted screen and rotarod assays, only by the intraperitoneal route. Hesperetin did not show any effects in vivo in mice in these models, but in vitro it displaced the [3H]DAMGO binding with low-affinity and inhibited inward currents through the expressed GIRK1/2 channels. Although hesperidin actions in vivo demonstrated to be mediated by an opioid mechanism of action, it failed to directly bind to and activate the MU-opioid receptor or produce any change on inward GIRK1/2 currents in vitro. However, it should be considered that hesperidin may be metabolized, possibly resulting in crucial changes in its biological activity. PMID- 21624390 TI - N-methyl-D-aspartate receptor antagonist MK-801 suppresses glial pro-inflammatory cytokine expression in morphine-tolerant rats. AB - Chronic opioid therapy induces tolerance and hyperalgesia, which hinders the efficacy of opioid treatment. Previous studies have shown that inhibition of neuroinflammation and glutamatergic receptor activation prevents the development of morphine tolerance. The aim of the present study was to examine whether N Methyl-D-aspartate receptors are involved in the regulation of chronic morphine induced neuroinflammation in morphine-tolerant rats. Morphine tolerance was induced in male Wistar rats by intrathecal infusion of morphine (15 MUg/h) for 5 days. Tail-flick latency was measured to estimate the antinociceptive effect of morphine. Morphine challenge (15 MUg, intrathecally) on day 5 at 3h after discontinuation of morphine infusion produced a significant antinociceptive effect in saline-infused rats, but not in morphine-tolerant rats. Pretreatment with MK-801 (20 MUg, intrathecally) 30 min before morphine challenge preserved its antinociceptive effect in morphine-tolerant rats. Morphine-tolerant rats expressed high levels of the pro-inflammatory cytokines interleukin-1beta, interleukin-6, and tumor necrosis factor-alpha and the increase in interleukin 1beta and interleukin-6, and tumor necrosis factor-alpha levels was prevented by MK-801 pre-treatment at both the protein and mRNA levels. The results show that a single dose of MK-801 reduces the increase in pro-inflammatory cytokines in the spinal cord, thus re-sensitizing neurons to the antinociceptive effect of morphine in morphine-tolerant rats. This study provides a piece of theoretical evidence that NMDA antagonist can be a therapeutic adjuvant in treating morphine tolerant patients for pain relief. PMID- 21624391 TI - Low dose, short-term rivastigmine administration does not affect neurocognition in methamphetamine dependent individuals. AB - Neurocognitive impairment is a well-documented consequence of methamphetamine addiction. Not surprising, methamphetamine-associated neurocognitive impairment has been identified as an important target of treatment. Thus, this study sought to determine whether rivastigmine, an acetylcholinesterase inhibitor and cognition enhancing agent, could improve neurocognitive performance in a sample of long-term, high-dose methamphetamine addicts who were not seeking treatment at the time of enrollment in the study. This double-blind, placebo-controlled study evaluated whether a daily dose 0, 3, or 6 mg of rivastigmine, administered over six consecutive days, would enhance performance on measures of attention/information processing speed, episodic memory, and executive/frontal lobe functioning relative to test performance at baseline. The results revealed that rivastigmine did not alter neurocognition in this cohort. There are a number of factors that may have mitigated the effects of rivastigmine in this particular study, including especially the short-term, low-dose treatment regimen utilized. The negative findings notwithstanding, the study serves as a springboard for future investigations that will examine whether other medications can alter neurocognition in methamphetamine dependent study participants. PMID- 21624392 TI - A high fructose diet does not affect amphetamine self-administration or spatial water maze learning and memory in female rats. AB - High energy diets can have a detrimental effect on brain plasticity. For example, a high fructose diet impairs spatial memory in male rats. The aim of the present study was to determine whether a high fructose diet impairs another form of learning and memory: drug reinforcement learning. Female Sprague-Dawley rats were fed a high fructose diet (60%) from weaning at postnatal day (PND) 21, then allowed to acquire lever-pressing maintained by intravenous (i.v.) amphetamine at PND 68, 109, or 165. Acquisition was tested on a fixed ratio one (FR1) schedule of reinforcement (0.025 mg/kg/infusion, 1h daily sessions, 10 sessions over 14 days), followed by testing for reinforcing efficacy on a progressive ratio (PR) schedule (0.025, 0.01, and 0.1mg/kg/infusion), 14 days of abstinence, and within session extinction and reinstatement tests. Subsequently, water maze acquisition and retention were tested in these subjects as well as a separate cohort tested in the water maze only. The diet had no effect on acquisition, reinforcing efficacy, extinction, or reinstatement of amphetamine seeking. Nor did the diet alter any measures of spatial memory. The high fructose diet did decrease body mass and increase relative liver and spleen mass, but did not affect plasma triglyceride concentrations consistently. Together with prior research on males, these results suggest that the metabolism of fructose and the effects of a high fructose diet on learning and memory may be sex-dependent. PMID- 21624393 TI - The antinociceptive potency of N-arachidonoyl-dopamine (NADA) and its interaction with endomorphin-1 at the spinal level. AB - The endogenous N-arachidonoyl-dopamine (NADA) activates both transient receptor potential vanilloid1 (TRPV1) and cannabinoid-1 (CB(1)) receptors. The goal of this study was to characterize the antinociceptive potential of NADA on inflammatory thermal hyperalgesia in rats at spinal level, and to determine its interaction with endomorphin-1 (EM) at the spinal level. The effects of NADA and EM on thermal hyperalgesia were evaluated in rats with a unilateral hind paw carrageenan-induced inflammation. Intrathecal injection of either EM (0.03-10 MUg) or NADA (1.5-50 MUg) caused dose-dependent antihyperalgesia, but NADA was 5.4 times less potent than EM. The antihyperalgesia caused by 15 MUg NADA was inhibited by the TRPV1 antagonist AMG9810, but not by CB(1) antagonist/inverse agonist AM 251, whereas the effect of 50 MUg NADA was decreased by both drugs. Co administration of EM with NADA in 1:15 and 1:50 ratios produced a short-lasting potentiation, but isobolographic analysis for the whole investigated period revealed additive interaction between the two endogenous ligands. The results show that both TRPV1 and CB(1) receptor activation play a substantial role in the antinociceptive effects of NADA at spinal level, while co-administration of NADA with EM did not show potentiation. PMID- 21624394 TI - The effect of neighborhood, socioeconomic status and a community-based program on multi-disease health screening in an Asian population: a controlled intervention study. AB - OBJECTIVES: We studied whether individual socioeconomic and neighborhood factors such as living in a poor community independently affected health screening participation. METHODS: We studied 3 blocks of public-rental flats (the poorer neighborhood) adjacent to 3 blocks of owner-occupied public housing (the better off neighborhood) in a precinct in Taman Jurong, Singapore. Demographic details and reasons for not having regular hypertension, diabetes mellitus, hyperlipidemia and colorectal cancer screening were collected from 2009 to 2010. An access-enhancing intervention was implemented in both neighborhoods to raise health screening rates. RESULTS: Participation rates for rental flats and owner occupied flats were 89.0% (356/400) and 70.2% (351/500) respectively. Living in a better-off neighborhood was independently associated with diabetes mellitus (66% vs. 35%, adjusted odds ratio (AOR)=2.12, p<0.01), hyperlipidemia (53% vs. 26%, AOR=4.34, p<0.01) and colorectal cancer screening (17% vs. 6%, AOR=15.43, p<0.01), as were individual socioeconomic factors such as employment, need for financial aid and household income. Uptake of all screening modalities significantly increased in the poorer neighborhood post-intervention (all p<0.05). Cost was cited more commonly as a barrier to health screening in the poorer neighborhood. CONCLUSION: Differing neighborhoods within one geographical location, as well as individual socioeconomic factors, were independently associated with differences in health screening. PMID- 21624395 TI - Structural analysis of alpha-helical proteins from wool using cysteine labelling and mass spectrometry. AB - A simple reduction/labelling/extraction protocol has been developed to fractionate cortical matrix proteins from filament proteins in wool. Through differential labelling of cysteine residues their relative accessibility in the wool fibre has been investigated. This has allowed the preliminary development of a map of the chemical functionality that is accessible within wool fibres under native conditions. Protein analyses of wool subjected to mechanical action, wet chemical permonosulphate/sulphite treatment and dry argon plasma treatment revealed that none of these detectably improved the accessibility of functional groups at the wool cortex. It is anticipated that this analytical method can be extended to improve the sensitivity and scope with which chemical functionality within native fibres can be mapped and lead to a better understanding of the potential limits/opportunities for fibre modification. PMID- 21624396 TI - Fucoidan from Sargassum sp. and Fucus vesiculosus reduces cell viability of lung carcinoma and melanoma cells in vitro and activates natural killer cells in mice in vivo. AB - Fucoidan is known to exhibit crucial biological activities, including anti-tumor activity. In this study, we examined the influence of crude fucoidan extracted from Sargassum sp. (MTA) and Fucus vesiculosus (SIG) on Lewis lung carcinoma cells (LCC) and melanoma B16 cells (MC). In vitro studies were performed using cell viability analysis and showed that SIG and MTA fucoidans significantly decreased the viable number of LCC and MC cells in a dose-response fashion. Histochemical staining showed morphological changes of melanoma B16 cells after exposure to fucoidan. The observed changes were indicative of crude fucoidan induced apoptosis. Male C57BL/6JJCL mice were subjected to daily i.p. injections over 4 days with either SIG or MTA fucoidan (50mg/kg body wt.). The cytolytic activity of natural killer (NK) cells was enhanced by crude fucoidan in a dose dependent manner as indicated by (51)Cr labeled YAC-1 target cell release. This study provides substantial indications that crude fucoidan exerts bioactive effects on lung and skin cancer model cells in vitro and induces enhanced natural killer cell activity in mice in vivo. PMID- 21624397 TI - Central effects of a local inflammation in three commonly used mouse strains with a different anxious phenotype. AB - As in humans, genetic background in rodents may influence a peculiar set of behavioural traits such as sensitivity to pain and stressors or anxiety-related behaviours. Therefore, we tested the hypothesis that mice with different genetic backgrounds [outbred (CD1), inbred (C57BL/6J) and hybrid (B6C3F1) adult male mice] display altered reactivity to pain, stress and anxiety related behaviours. We demonstrated that B6C3F1 mice displayed the more anxious phenotype with respect to C57BL/6J or CD1 animals, with the latter being the less anxious strain when tested in an open field and on an elevated plus maze. No difference was observed across strains in thermal sensitivity to a radiant heat source. Mice were then treated with a sub-plantar injection of the inflammatory agent Complete Freund's Adjuvant (CFA), 24h later they were hyperalgesic with respect to saline exposed animals, irrespective of strain. We then measured intra-strain differences and CFA-induced inter-strain effects on the expression of various genes with a recognized role in pain and anxiety: BDNF, IL-6, IL-1beta, IL-18 and NMDA receptor subunits in the mouse thalamus, hippocampus and hypothalamus. The more anxious phenotype observed in B6C3F1 hybrid mice displayed lower levels of BDNF mRNA in the hippocampus and hypothalamus when compared to outbred CD1 and C57BL/6J inbred mice. CFA led to a general decrease in central gene expression of the evaluated targets especially in CD1 mice, while BDNF hypothalamic downregulation stands out as a common effect of CFA in all three strains evaluated. PMID- 21624398 TI - Adult neurogenesis: optimizing hippocampal function to suit the environment. AB - Numerous studies have attempted to determine the function of adult neurogenesis in the hippocampus using methods to deplete new neurons and examine changes in behaviors associated with this brain region. This approach has produced a set of findings that, although not entirely consistent, suggest new neurons are associated with improved learning and reduced anxiety. This paper attempts to synthesize some of these findings into a model that proposes adaptive significance to experience-dependent alterations in new neuron formation. We suggest that the modulation of adult neurogenesis, as well as of the microcircuitry associated with new neurons, by experience prepares the hippocampus to meet the specific demands of an environment that is predictably similar to one that existed previously. Reduced neurogenesis that occurs with persistent exposure to a high threat environment produces a hippocampus that is more likely to respond with behavior that maximizes the chance of survival. Conversely, enhanced neurogenesis that occurs with continual exposure to a rewarding environment leads to behavior that optimizes the chances of successful reproduction. The persistence of this form of plasticity throughout adulthood may provide the neural substrate for adaptive responding to both stable and dynamic environmental conditions. PMID- 21624399 TI - A transcriptomic study on the pepsin-activated infective larvae of Angiostrongylus cantonensis. AB - To investigate the pepsin-activated infective (third-stage) larvae of Angiostrongylus cantonensis at the transcriptomic level, 1496 ESTs were generated from a cDNA library and clustered into 161 contigs and 757 singletons. Among these unigenes, 54.5% had significant sequence homology with known proteins. The most abundantly expressed transcripts were cathepsin B-like cysteine protease 1 and 2, metalloprotease I, metalloprotease 1 precursor, and extracellular superoxide dismutase. Protein complex was the most common Gene Ontology classification within the 'cellular component' category, embryonic development ending in birth or egg hatching within 'biological process', and protein binding within 'molecular function'. Moreover, 280 clusters were mapped to 158 KEGG pathways and 134 had unique EC numbers. These findings suggest that treatment with pepsin-HCl not only digests the tissues of the snail host but also activates the infective larvae. PMID- 21624400 TI - Quantitation of Helicobacter pylori ureC gene and its comparison with different diagnostic techniques and gastric histopathology. AB - Numerous diagnostic assays for Helicobacter pylori detection are available. However, these techniques have their own advantages as well as limitations. Here we tried to develop a real-time quantitative (Q) PCR assay to measure ureC copy number to detect H. pylori, based on the fact that there is only one copy of the ureC gene per bacterium. We enrolled 120 adult patients [non-ulcer dyspepsia (NUD) 60, peptic ulcer disease (PUD) 20, gastric cancer (GC) 40] undergoing upper gastrointestinal endoscopies. During each endoscopic examination, antral biopsies from normal region of the antrum were obtained and subjected to the following tests: RUT, culture, histopathology, H. pylori-specific ureC PCR and ureC Q-PCR. Calculation of H. pylori copy number was based on the standard curve generated using 10-fold dilutions of DNA extracted from the H. pylori control strain varying from 10(5) to 10(1) copies. The prevalence of H. pylori infection in our study population was 54% with no significant difference among disease and control population. The sensitivity of Q-PCR was found to be 100% which was highest among all diagnostic tests. The established Q-PCR is around 10 times more sensitive than the conventional PCR method. The copy number of H. pylori DNA was significantly increased when overall gastritis, H. pylori density, chronic inflammation and intestinal metaplasia were present. In summary, we developed a rapid and sensitive Q-PCR method for detecting H. pylori. This technique offers a significant improvement over other available methods for detecting H. pylori in clinical and research samples. PMID- 21624401 TI - Differential expression of voltage-gated K+ currents in medial septum/diagonal band complex neurons exhibiting distinct firing phenotypes. AB - The medial septum/diagonal band complex (MSDB) controls hippocampal excitability, rhythms and plastic processes. Medial septal neuronal populations display heterogeneous firing patterns. In addition, some of these populations degenerate during age-related disorders (e.g. cholinergic neurons). Thus, it is particularly important to examine the intrinsic properties of theses neurons in order to create new agents that effectively modulate hippocampal excitability and enhance memory processes. Here, we have examined the properties of voltage-gated, K(+) currents in electrophysiologically-identified neurons. These neurons were taken from young rat brain slices containing the MS/DB complex. Whole-cell, patch recordings of outward currents were obtained from slow firing, fast-spiking, regular-firing and burst-firing neurons. Slow firing neurons showed depolarization-activated K(+) current peaks and densities larger than in other neuronal subtypes. Slow firing total current exhibited an inactivating A-type current component that activates at subthreshold depolarization and was reliably blocked by high concentrations of 4-AP. In addition, slow firing neurons expressed a low-threshold delayed rectifier K(+) current component with slow inactivation and intermediate sensitivity to tetraethylammonium. Fast-spiking neurons exhibited the smaller I(K) and I(A) current densities. Burst and regular firing neurons displayed an intermediate firing phenotype with I(K) and I(A) current densities that were larger than the ones observed in fast-spiking neurons but smaller than the ones observed in slow-firing neurons. In addition, the prevalence of each current differed among electrophysiological groups with slow firing and regular firing neurons expressing mostly I(A) and fast spiking and bursting neurons exhibiting mostly delayer rectifier K(+) currents with only minimal contributions of the I(A). The pharmacological or genetic modulations of these currents constitute an important target for the treatment of age-related disorders. PMID- 21624402 TI - Presynaptic nicotinic potentiation of a frog retinotectal transmission evoked by discharge of a single retina ganglion cell. AB - It was demonstrated in our previous studies of the frog retinotectal transmission that retinotectal synaptic potentials are enhanced by a factor of 1.5 due to the tonic presynaptic nicotinic potentiation, caused by the ambient level of the acetylcholine in the frog tectum. Furthermore, the results of those studies have indicated that the mechanism of the nicotinic potentiation is only partially exploited, because the application of the cholinergic agonist had increased the retinotectal transmission more than 2 times above the level of the tonic potentiation. The purpose of the present study was to explore this additional potentiation. We have shown that: (1) Bursts of 4-10 action potentials of a frog retina ganglion cell gave rise to an increase (phasic potentiation) of the retinotectal transmission 1.4-2.2 times, depending on the burst strength, that lasted tens of seconds. (2) This increase has been mediated through the presynaptic nicotinic acetylcholine receptors activated by the endogenous acetylcholine released into the tectum during relatively strong bursts of the retina ganglion cell. (3) Two types of the nicotinic acetylcholine receptors are co-localized in the presynaptic terminals of the individual retinotectal input to the tectum layer F--high-affinity (tonic) and low-affinity (phasic) nicotinic receptors. PMID- 21624403 TI - Protein aggregate spreading in neurodegenerative diseases: problems and perspectives. AB - Progressive accumulation of specific protein aggregates is a defining feature of many major neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, fronto-temporal dementia, Huntington's disease, and Creutzfeldt-Jakob disease (CJD). Findings from several recent studies have suggested that aggregation-prone proteins, such as tau, alpha-synuclein, polyglutamine containing proteins, and amyloid-beta, can spread to other cells and brain regions, a phenomenon considered unique to prion disorders, such as CJD and bovine spongiform encephalopathy. Cell-to-cell propagation of protein aggregates may be the general underlying principle for progressive deterioration of neurodegenerative diseases. This may also have significant implications in cell replacement therapies, as evidenced by the propagation of alpha-synuclein aggregates from host to grafted cells in long-term transplants in Parkinson's patients. Here, we review recent progress in protein aggregate propagation in experimental model systems and discuss outstanding questions and future perspectives. Understanding the mechanisms of this pathological spreading may open the way to unique opportunities for development of diagnostic techniques and novel therapies for protein misfolding-associated neurodegenerative diseases. PMID- 21624404 TI - Differential vasoactive effects of oestrogen, oestrogen receptor agonists and selective oestrogen receptor modulators in rat middle cerebral artery. AB - Cerebrovascular disorders are less common in pre-menopausal than post-menopausal women and in females than males. This protection may be due, in part at least, to direct effects of oestrogens on blood vessels. Oestrogen's vasodilatory mechanisms have been reported to be via the endothelium, vascular smooth muscle and extracellular matrix, depending on the vascular bed studied. Herein we investigated the vasoactive effects of oestrogen, oestrogen receptor (ER) and GPR30 agonists and selective ER modulators (SERMs) in the rat middle cerebral artery(MCA), an artery affected in focal ischaemia. MCAs isolated from male Sprague Dawley rats were mounted on a wire myograph. Concentration response curves were constructed to 17beta-oestradiol, ERalpha agonist-PPT, ERbeta agonist DPN, GPR30 agonist-G1 and novel SERMs (LY362321 and LY2120310) in pre-constricted vessels, in the presence and absence of endothelium, blocking agents for nitric oxide synthase (L-NAME), classic ER antagonist (ICI182,780) or plasma membrane specific ERalpha (ERalpha-36) antibody. 17beta-oestradiol induced rapid vasorelaxation of the MCA which was not affected by endothelium removal, L-NAME or ICI182,780. Vasorelaxation was mimicked by PPT, DPN and G1 but not by the SERMs. Using ERalpha-36 antibody, effects of oestrogen were partially blocked. PPT had a greater vasorelaxation, while DPN and G1 had a lesser effect than 17beta-oestradiol. These findings indicate that activation of plasma membrane bound ERalpha, beta and GPR30 elicits rapid, endothelial-nitric oxide-independent relaxation of the rat MCA. PMID- 21624405 TI - Analysis of the cell cycle regulatory protein (E2F1) after infection of cultured cells with bovine herpesvirus 1 (BHV-1) or herpes simplex virus type 1 (HSV-1). AB - The E2F family of cellular transcription factors controls cell cycle progression and cell death. During cell cycle progression, activated cyclin-dependent kinases phosphorylate the retinoblastoma (Rb) protein, causing the release and activation of E2F family members. Previous studies demonstrated that bovine herpes virus 1 (BHV-1) productive infection increases E2F1 protein levels, the bICP0 early promoter is activated more than 100 fold by E2F1 or E2F2, and silencing E2F1 reduced the efficiency of productive infection. In this study, the effect of herpes simplex virus type 1 (HSV-1) productive infection on E2F protein levels and regulation of E2F dependent transcription was compared to BHV-1 infection in the same permissive cell line, rabbit skin (RS) cells. Silencing E2F1 with a specific siRNA reduced HSV-1 productive infection approximately 10 fold in RS cells, and total E2F1 protein levels increased during productive infection. In contrast to RS cells infected with BHV-1, a fraction of total E2F1 protein was localized to the cytoplasm in HSV-1 infected RS cells. Furthermore, E2F1 did not efficiently trans-activate the HSV-1 ICP0 or ICP4 promoter. When RS cells were transfected with an E2F reporter construct or the cyclin D1 promoter and then infected with BHV-1, promoter activity increased after infection. In contrast, HSV-1 infection of RS cells had little effect on E2F dependent transcription and cyclin D1 promoter activity was reduced. In summary, these studies indicated that silencing E2F1 reduced the efficiency of HSV-1 and BHV-1 productive infection. However, only BHV-1 productive infection induced E2F dependent transcription. PMID- 21624406 TI - Development of tripartite polyion micelles for efficient peptide delivery into dendritic cells without altering their plasticity. AB - For many years, a great deal of interest has been focusing on the optimization of peptide presentation by dendritic cells (DCs) using peptide-encapsulated particles, in order to enhance the immune response. Nowadays, DCs are also known to be involved in peripheral tolerance, inducing anergy or regulatory T lymphocytes. To preserve the plasticity of DCs, we formulated non-cytotoxic pH sensitive polyion complex micelles based on an original tripartite association of polymethacrylic acid-b-polyethylene oxide, poly-L-lysine and fluorescent-peptide: OVAFITC peptide, as a model drug. We demonstrated that the OVAFITC peptide was successfully entrapped into the micelles, released into DC endosomes thanks to the pH-sensitivity property of the micelles, and efficiently loaded onto MHC class II molecules. The phenotype as well as the cytokinic secretion profile of the mature and immature DCs loaded with peptide-encapsulated micelles was unaltered by the tripartite polyion micelles. The efficient loading of the peptide by immature and mature DCs was shown by the in vitro proliferation of OVA specific transgenic T cells. Therefore, the present results show that the tripartite polyion complex micelles can be used as efficient peptide vectors immunogically inert for ex vivo DCs engineering without modifying their intrinsic immune plasticity. PMID- 21624407 TI - Compliance with Mediterranean Diet Quality Index (KIDMED) and nutrition knowledge levels in adolescents. A case study from Turkey. AB - Adopting an eating pattern complying with the Mediterranean diet not only decreases body fat mass and obesity risk, but also reduces development of various health problems. This study investigated the nutritional awareness and diet quality Mediterranean Diet Quality Index (KIDMED) of Turkish adolescents. The study was conducted with 890 voluntary participants (464 boys and 426 girls) aged 10-14 years. A questionnaire form was used to learn demographic characteristics of the participants. Participants' nutritional awareness was determined through a 20-item knowledge form and their nutritional habits through a 16-item Mediterranean Diet Quality Index (KIDMED). The average nutritional knowledge score was X=82.22 +/- 0.42. Results indicated that 17.9% of the participants had a low quality diet (<= 3 points), 59.2% had a mid-quality/needs-improvement diet (4-7 points) and 22.9% had an optimal quality diet (>= 8 points). The study results showed that the subjects' diet quality was low and that their nutrition knowledge levels were related to their nutritional habits. PMID- 21624408 TI - The new kappa-KTx 2.5 from the scorpion Opisthacanthus cayaporum. AB - The kappa-KTx family of peptides, which is the newest K+-channel blocker family from scorpion venom, is present in scorpions from the families Scorpionidae and Liochelidae. Differently from the other scorpion KTx families, the three dimensional structure of the known kappa-KTxs toxins is formed by two parallel alpha-helices linked by two disulfide bridges. Here, the characterization of a new kappa-KTx peptide, designated kappa-KTx 2.5, derived from the Liochelidae scorpion Opisthacanthus cayaporum, is described. This peptide was purified by HPLC and found to be identical to OcyC8, a predicted mature sequence precursor (UniProtKB C5J89) previously described by our group. The peptide was chemically synthesized and the circular dichroism (CD) spectra of both, native and synthetic, conducted at different temperatures in water and water/trifluoroethanol (TFE), showed a predominance of alpha-helices. The kappa KTx 2.5 is heat stable and was shown to be a blocker of K+-currents on hKv1.1, and hKv1.4, with higher affinity for Kv1.4 channels (IC50= 71 MUM). Similarly to the other kappa-KTxs, the blockade of K+-channels occurred at micromolar concentrations, leading to uncertainness about their proper molecular target, and consequently their pharmacologic effect. In order to test other targets, kappa KTx2.5 was tested on other K+-channels, on Na+-channels, on bacterial growth and on smooth muscle tissue, a known assay to identify possible bradykinin potentiating peptides, due to the presence of two contiguous prolines at the C terminal sequence. It has no effect on the targets used except on hKv1.1, and hKv1.4 expressed in Chinese hamster ovary cells. Since the only plausible function found for kappa-KTx2.5 seems to be the blockade of K+-channels, a discussion regarding the analysis of structure-function relationships is included in this communication, based on sequence alignments of members of the kappa-KTx toxin family, and on computational simulation of a structural model of the kappa KTx2.5-Kv1.2 complex. PMID- 21624409 TI - Selective activation of metabotropic glutamate receptor 7 induces inhibition of cellular proliferation and promotes astrocyte differentiation of ventral mesencephalon human neural stem/progenitor cells. AB - Expression of group III metabotropic glutamate receptors (mGluR) was established by RT-PCR and immunocytochemistry on a cultured clonal human neural stem/progenitor cell (hNSPC) line derived from fetal ventral mesencephalon (VM). Selective activation of these receptors by the group III mGluR agonist L-(+)-2 amino-4-phosphonobutyric acid (L-AP4) prevented increases in cAMP levels following forskolin stimulation, suggesting these receptors are coupled to their canonical G-protein coupled signal transduction pathway. Tonic exposure of undifferentiated cultures to L-AP4 resulted in a decrease in cellular metabolism and proliferation in the absence of toxicity, as measured by MTT and LDH assays, in a dose-dependent manner. This was confirmed by a reduction in BrdU incorporation into nuclear DNA, suggestive of an anti-proliferative effect of L AP4. This effect was rescued by co-addition of the broad-spectrum group III mGluR competitive antagonist (RS)-a-cyclopropyl-4-phosphonophenylglycine (CPPG), demonstrating a receptor-mediated mechanism, but not mimicked by application of the cell permeable cAMP analogue dibutyrl cAMP (db-cAMP). The potency of these effects of L-AP4 indicates that this is an mGlu7 subtype-mediated effect. Tonic exposure of undifferentiated cultures to the mGlu7 selective allosteric agonist N,N'-bis(diphenylmethyl)-1,2-ethanediamine dihydrochloride (AMN082), but not the mGlu4 selective allosteric agonist (+/-)-cis-2-(3,5 dicholorphenylcarbamoyl)cyclohexanecarboxylic acid (VU0155041), or the mGlu8 selective agonist (S)-3,4-dicarboxyphenylglycine ((S)-3,4-DCPG) resulted in an identical anti-proliferative effect to L-AP4, confirming the involvement of the mGlu7 subtype. In differentiating cultures, tonic exposure to L-AP4 or AMN082 resulted in a significant shift towards an astrocyte cell fate. The mGlu7 receptor therefore provides a new opportunity to influence the proliferation and differentiation of ventral mesencephalon-derived hNSPC. PMID- 21624410 TI - Extended periods of neural induction and propagation of embryonic stem cell derived neural progenitors with EGF and FGF2 enhances Lmx1a expression and neurogenic potential. AB - Neural stem (NS) cells are multipotent cells defined by their capacity to proliferate and differentiate into all neuronal and glial phenotypes. NS cells can be obtained from specific regions of the adult brain, or generated from embryonic stem cells (ESCs). NS cells differentiate into neural progenitor (NP) cells and subsequently neural precursors, as transient steps towards terminal differentiation into specific mature neuronal or glial phenotypes. When cultured in EGF and FGF2, ESC-derived NS cells have been reported to be stable and multipotent. Conditions that enable differentiation of NS cells through the committed progenitor and precursor stages to specific neuronal subtypes have not been fully established. In this study we investigated, using Lmx1a reporter ESCs, whether the length of neural induction (NI) dictated the phenotypic potential of cultures of ESC-derived NS cells or NP cells. Following 4, 7 or 10 day periods of NI, ESCs in monolayer culture were harvested and cultured as neurospheres, prior to replating as monolayer cultures for several passages in EGF and FGF2. The NS/NP cultures were then directed towards mature neuronal fates over 16-17 days. 4 and 7-day NS cell cultures could not be differentiated towards dopaminergic, serotonergic or cholinergic fates as determined by the absence of tyrosine hydroxylase, 5-HT or choline acetyltransferase (ChAT) immunolabelling. In contrast NS/NP cultures derived after 10 days of NI were able to generate tyrosine hydroxylase and 5-HT positive neurons (24+/-6 and 13+/-1% of the betaIII tubulin positive population, respectively, n=3). Our data suggest that extended periods of neural induction enhanced the potential of mouse ESC-derived NS/NP cells to generate specific subtypes of neurons. NS/NP cells derived after shorter periods of NI appeared to be lineage-restricted in relation to the neuronal subtypes observed after removal of EGF. PMID- 21624411 TI - Immunogenicity and safety of inactivated influenza vaccines in primed populations: a systematic literature review and meta-analysis. AB - Several inactivated influenza vaccine formulations for systemic administration in man are currently available for annual (seasonal) immunization: split virus and subunit (either plain-aqueous, or virosomal, or adjuvanted by MF59). From a literature search covering the period 1978-2009, 33 articles could be identified, which described randomized clinical trials comparing at least two of the four vaccine formulations with respect to serum hemagglutination inhibition (HI) antibody response, local and systemic vaccine reactions and serious adverse events after vaccination, and employing seasonal vaccine components and doses. In total, 9121 vaccinees of all ages, either healthy or with underlying diseases, were involved. Most vaccinees were primed or had been vaccinated in previous years. For immunogenicity, homologous post-vaccination geometric mean HI titers (GMTs) were analyzed by a random effects model for continuous data. Unreported standard deviations (SD) were addressed by imputing assumed SD-values. Age and health state of the vaccinees appeared to have little influence on the outcome. The immunogenicity of split, aqueous and virosomal subunit formulations were similar, with geometric mean ratio values (GMR, quotient of paired GMT-values) varying around one (0.93-1.24). The MF59-adjuvanted subunit vaccine induced, on average, larger antibody titers than the non-adjuvanted vaccine formulations, but the absolute increase was small (GMR-values varying between 1.25 and 1.40). Vaccine reactions were analyzed using a random effects model for binary data. Local and systemic reactogenicity was similar among non-adjuvanted formulations. The adjuvanted subunit formulation was more frequently associated with local reactions than the non-adjuvanted formulations (rate ratio: 2.12, significant). Systemic reactions were similar among all vaccine formulations. The original articles emphasized the mild and transient character of the vaccine reactions and the absence of serious vaccine-related adverse events. This adequate amount of evidence led to the conclusion that all the currently available inactivated influenza vaccine formulations are safe, well tolerated and similarly effective to control seasonal influenza outbreaks across primed populations and age ranges. PMID- 21624412 TI - Persistence of antibodies in 4-8 year old Austrian children after vaccination with hexavalent DTaP-HBV-IPV/Hib and MMR vaccines. AB - To determine the proficiency of the Austrian childhood vaccination schedule to induce long lasting seroprotection against vaccine preventable diseases a seroepidemiological study in 348 children between four and eight years of age was conducted. Antibodies against diphtheria, tetanus, pertussis, hepatitis B, measles, mumps and rubella antigens were assessed in children, who had been vaccinated with hexavalent DTaP-HBV-IPV/Hib vaccines at three, four, five months and in the second year of life and/or MMR vaccines in the second year of life at least once, but mostly twice. High seroprotection rates (SPRs) were detected for tetanus (96%) and measles (90%). SPRs regarding diphtheria and mumps were 81% and 72%, respectively. Rubella-SPRs were 68% in females and 58% in males. Hepatitis B antibody levels >=10 mIU/mL were present in 52%; antibodies against pertussis were detected in 27% of the children. SPRs for measles and rubella depended on the interval since last vaccination; mumps-antibodies were significantly lower after one MMR-vaccination only. Antibodies against diphtheria, tetanus and pertussis depended on the interval since last vaccination while HBs-antibodies did not. The low levels of antibodies 1-7 years after vaccination against pertussis, rubella and mumps after only one vaccination should be considered when recommending new vaccination schedules. PMID- 21624413 TI - Improvement of H5N1 influenza vaccine viruses: influence of internal gene segments of avian and human origin on production and hemagglutinin content. AB - The H5N1-clade 1 influenza vaccine strain NIBRG-14 produces exceptionally low amounts of antigen, a problem recently encountered also for initial pandemic H1N1 2009 vaccine seeds. Here, we report on a strategy that may contribute to overcome this obstacle. Influenza vaccine viruses usually consist of two segments coding for the antigenic HA and NA proteins of a wild-type strain and the six residual internal gene segments of the vaccine donor strain A/PR/8/34 (PR8). To enhance the antigen yield from H5N1 vaccine virus we generated by reverse genetics a set of PR8-based reassortant viruses expressing the HA and NA segments of the prototypic strain A/Vietnam/1203/2004 and additional replacements of the internal M or PB1 genes of PR8. The reassortants were compared to the parental PR8 and H5N1 viruses in terms of growth in embryonated chicken eggs and the amount of incorporated antigenic HA protein. Compared to NIBRG-14, three out of six viruses displayed an increased replication in embryonated chicken eggs and higher HA content that was also maintained after ether/detergent extraction of virions. Electron microscopic analysis showed that the reassortment hardly affected particle shape and size. Two selected H5N1 reassortant viruses were investigated concerning their pathogenicity in ferrets and found to behave as low pathogenic as the PR8 donor strain. In conclusion, this study shows that replication and antigen content of PR8-derived H5N1 influenza vaccine viruses can be improved by incorporation of heterologous internal gene segments without compromising their attenuated character. PMID- 21624414 TI - Antigen-specific enhancement of natural human IgG antibodies to phosphatidylcholine, phosphatidylglycerol, phosphatidylinositol-4-phosphate, cholesterol, and lipid A by a liposomal vaccine containing lipid A. AB - Natural IgG antibodies (NA) to lipids are ubiquitously distributed in sera of healthy humans and are believed to serve beneficial functions. Although NA to lipids generally exhibit germ line or near germ line binding specificities, the antibodies commonly increase transiently in the acute phases of most, if not all, infectious diseases and may serve as a first line of defense. In order to determine whether similar anti-lipid antibodies can be induced by a vaccine in humans, we examined stored sera obtained from volunteers who had previously received a candidate vaccine to Plasmodium falciparum. The vaccine had consisted of liposomes that contained both the recombinant protein antigen and also contained monophosphoryl lipid A (MPLA) as an adjuvant. All of the pre-immune sera contained NA to one or more of the liposomal lipids in the vaccine: dimyristol phosphatidylcholine (DMPC), dimyristoyl phosphatidylglycerol (DMPG), cholesterol, and MPLA. After initial immunization, followed by a boost, increased levels of IgG antibodies to all of the liposomal lipids, especially DMPG and MPLA, were observed by ELISA. Antibodies to phosphatidylinositol-4-phosphate (PIP) above the normal pre-immune NA to PIP were also observed. Although PIP was not present in the immunizing liposomes, based on the adsorption of anti-PIP antibodies by DMPG the anti-PIP antibodies were thought to represent cross reacting anti-DMPG antibodies. The immune response was apparently antigen specific in that NA to unrelated lipids, other than PIP, that were not present in the liposomes, galactosyl ceramide and ganglioside GM1, were not increased by the immunization. We conclude that antibodies to DMPC, DMPG, PIP, cholesterol, and MPLA can be induced in humans by immunization with liposomes containing MPLA. PMID- 21624415 TI - Development of live attenuated Bordetella pertussis strains expressing the universal influenza vaccine candidate M2e. AB - The attenuated Bordetella pertussis BPZE1 vaccine strain represents an attractive platform for the delivery of heterologous vaccine candidates via the nasal route. The filamentous hemagglutinin (FHA) has been used to secrete or expose the foreign antigens at the bacterial surface. In this study, one, two and three copies of the Cys-containing ectodomain of matrix protein 2 (M2e) from influenza A virus were genetically fused to full length FHA and expressed in BPZE1. The secretion efficacy of the FHA-(M2e)(1,2,3) chimera in the extracellular milieu and the ability of the recombinant bacteria to colonize the mouse lungs inversely correlated with the number of M2e copies fused to FHA. Nevertheless FHA-(M2e)(3) producing bacteria (BPLR3) triggered the highest systemic anti-M2e antibody response upon nasal administration to BALB/c mice. Nasal immunization with BPLR3 bacteria resulted in a significant reduction in the viral loads upon challenge with H1N1/PR8 influenza A virus, but did not improve the survival rate compared to BPZE1-immunized mice. Furthermore, since previous work reported that disulfide bond formation in Cys-containing passenger antigens affects the secretion efficacy of the FHA chimera, the dsbA gene encoding a periplasmic disulfide isomerase was deleted in the FHA-(M2e)(3)-producing strain. Despite improving significantly the secretion efficacy of the FHA-(M2e)(3) chimera, the dsbA deletion did not result in higher anti-M2e antibody titers in mice, due to impaired bacterial fitness and colonization ability. PMID- 21624416 TI - Safety and immunogenicity of a recombinant M2e-flagellin influenza vaccine (STF2.4xM2e) in healthy adults. AB - BACKGROUND: The ectodomain of matrix protein 2 (M2e) is a promising candidate for a broadly protective influenza A vaccine because it is highly conserved and antibodies to M2e are protective in animal models. STF2.4xM2e (VAX102) is a recombinant fusion protein that links four tandem copies of the M2e antigen to Salmonella typhimurium flagellin, a TLR5 ligand used as an adjuvant. The objectives of this first-in-human study were to assess the safety and immunogenicity of VAX102 given as a prime-boost regimen to healthy adults. METHODS: Sixty subjects 18-49 years old were enrolled in a multicenter, double blind, randomized, placebo-controlled trial (Study 1). Based on pre-clincial data, initial design included doses starting at 10 MUg, with an escalation plan. After reactogenicity was noted at the 10 MUg dose, the trial was redesigned to evaluate 0.3, 1.0, and 3 MUg doses. Following this study, 16 subjects were enrolled in Study 2, an open label, low dose study, to evaluate doses of 0.03 and 0.1 MUg. In both trials, vaccine or placebo was given intramuscularly (i.m.) at 0 and 28 days. Clinical and laboratory safety assessments took place 1 and 7 days after immunization. Immune responses to M2e and flagellin were assessed by ELISA at 7, 14 and 28 days after each dose. Seroconversion was defined as a serum IgG anti-M2e antibody value >=0.174 MUg/ml and a fourfold rise in concentration. RESULTS: Doses of 0.03-1 MUg were safe and well tolerated in all subjects. Doses of 0.03 and 0.1 MUg produced limited immunogenicity (38% and 75% respectively), after the second dose of vaccine. Doses of 0.3 and 1.0 MUg were immunogenic in 18 (75%) of 24 vaccinees after the first dose and 23 (96%) after the second dose. In the 1.0 MUg group, the geometric mean M2e antibody concentration was 0.4 MUg/ml after the first dose and 1.7 MUg/ml after the booster dose. M2e antibody concentrations and seroconversion rates were not significantly different at higher doses (p>0.05). Immune response to flagellin was robust but did not appear to interfere with M2e antibody responses after the booster dose. Following the first injection of VAX102 at higher doses (3 and 10 MUg), self-limited but severe symptoms were noted in some subjects and were associated with elevated levels of C-reactive protein. Although not directly measured, this reaction was believed to be mediated by cytokine release. CONCLUSIONS: VAX102 was safe and induced high antibody levels to M2e at 0.3 and 1.0 MUg doses. The TLR5 ligand, S. typhimurium flagellin, is a novel approach to adjuvant-like activity through activation of innate immunity, and when fused to multiple copies of the M2e protein, the vaccine was able to induce a fourfold rise in antibody in humans, to a previously non-immunogenic, highly-conserved portion of the influenza virus. Clinical correlates of protection that may be afforded by M2e antibody in humans are a future focus of investigation. PMID- 21624417 TI - Difficulties in the prevention of cervical cancer: adults' attitudes towards HPV vaccination 3 years after introducing the vaccine in Hungary. AB - Cervical cancer is one of the most prevalent gynaecological malignancies worldwide. The Hungarian incidence and mortality of this disease take the 4th-5th places within the European Union. A survey including 785 male and female adults was conducted to assess the knowledge and attitudes concerning HPV vaccination. We focused on the difficulties of the primary and secondary prevention of cervical cancer and examined some potential sociodemographic predictors of HPV vaccine acceptability. Our findings have identified some important issues like: incomplete knowledge, intense distrust and financial concerns. Almost half of the college students (45.6%) are unaware of HPV infections. We confirmed previous findings that older age and female gender correlates with better knowledge on STDs, including HPV. We found that greater exposure to health information comes with better knowledge and more positive attitudes towards vaccination. One quarter of survey respondents do not believe that cervical cancer may be prevented by vaccination. More than half of the adults do not trust national health care system and the preparedness of Hungarian doctors. General attitudes towards vaccination are broadly positive, 80% of survey participants had expressed desire towards HPV vaccination, however if there was a need to pay for the vaccination the willingness would decrease by half. Primary prevention through HPV-focused educational programs, clear communication and financial support would be important for public health to reduce the high incidence and mortality of cervical cancer in Hungary in the future. PMID- 21624418 TI - Marked enhancement of the immune response to BioThrax(r) (Anthrax Vaccine Adsorbed) by the TLR9 agonist CPG 7909 in healthy volunteers. AB - Immunization with BioThrax((r)) (Anthrax Vaccine Adsorbed) is a safe and effective means of preventing anthrax. Animal studies have demonstrated that the addition of CpG DNA adjuvants to BioThrax can markedly increase the immunogenicity of the vaccine, increasing both serum anti-protective antigen (PA) antibody and anthrax toxin-neutralizing antibody (TNA) concentrations. The immune response to CpG-adjuvanted BioThrax in animals was not only stronger, but was also more rapid and led to higher levels of protection in spore challenge models. The B-class CpG DNA adjuvant CPG 7909, a 24-base synthetic, single-strand oligodeoxynucleotide, was evaluated for its safety profile and adjuvant properties in a Phase 1 clinical trial. A double-blind study was performed in which 69 healthy subjects, age 18-45 years, were randomized to receive three doses of either: (1) BioThrax alone, (2) 1 mg of CPG 7909 alone or (3) BioThrax plus 1 mg of CPG 7909, all given intramuscularly on study days 0, 14 and 28. Subjects were monitored for IgG to PA by ELISA and for TNA titers through study day 56 and for safety through month 6. CPG 7909 increased the antibody response by 6-8-fold at peak, and accelerated the response by 3 weeks compared to the response seen in subjects vaccinated with BioThrax alone. No serious adverse events related to study agents were reported, and the combination was considered to be reasonably well tolerated. The marked acceleration and enhancement of the immune response seen by combining BioThrax and CPG 7909 offers the potential to shorten the course of immunization and reduce the time to protection, and may be particularly useful in the setting of post-exposure prophylaxis. PMID- 21624419 TI - The optimal number of routine vaccines to order at health clinics in low or middle income countries. AB - In a low or middle income country, determining the correct number of routine vaccines to order at a health clinic can be difficult, especially given the variability in the number of patients arriving, minimal vaccination days and resource (e.g., information technology and refrigerator space) constraints. We developed a spreadsheet model to determine the potential impact of different ordering policies, basing orders on the arrival rates seen in the previous 1, 3, 6, or 12 sessions, or on long-term historical averages (where these might be available) along with various buffer stock levels (range: 5-50%). Experiments varied patient arrival rates (mean range: 1-30 per session), arrival rate distributions (Poisson, Normal, and Uniform) and vaccine vial sizes (range: 1 dose to 10-dose vials). It was found that when the number of doses per vial is small and the expected number of patients is low, the ordering policy has a more significant impact on the ability to meet demand. Using data from more prior sessions to determine arrival rates generally equates to a better ability to meet demand, although the marginal benefit is relatively small after more than 6 sessions are averaged. As expected, the addition of more buffer is helpful in obtaining better performance; however, this advantage also has notable diminishing returns. In general, the long-term demand rate, the vial sizes of the vaccines used and the method of determining the patient arrival rate all have an effect on the ability of a clinic to maximize the demand that is met. PMID- 21624420 TI - Midwife attitudes: an important determinant of maternal postpartum pertussis booster vaccination. AB - BACKGROUND: The study was designed to determine the feasibility of implementing routine dTpa vaccination in the maternity ward to new mothers and to assess midwives' attitudes toward pertussis booster vaccination, their perceived susceptibility and severity of pertussis in their patients' communities, the perceived barriers and benefits of their patients' vaccinations, and their cues to action and self-efficacy in delivering the vaccine. METHOD: A self-completed questionnaire was developed to evaluate constructs of the Health Belief Model as well as to measure midwife demographic information. Questionnaires were completed by midwives during in-services at both a public hospital and a private hospital in New South Wales, Australia. RESULTS: Midwives who perceived ease in integrating booster vaccination into their workload were more likely to have high self-efficacy in delivering booster vaccination, measured through perceived importance of the role as part of their job (r = .449, p<.01), perceived confidence in delivering vaccination as part of their role (r = .608, p<.01), and perceived sufficient level of skills to deliver booster vaccination (r = .528, p<.01). CONCLUSIONS: These results suggest that, of the factors measured, the most important to midwives in terms of providing pertussis booster vaccination to mothers was their own perceived self-efficacy of providing the vaccination. To increase midwives' desire and confidence to provide pertussis booster to mothers, educational materials and skills workshops could be offered. PMID- 21624421 TI - Cost-effectiveness of rotavirus vaccination in Bolivia from the state perspective. AB - BACKGROUND: In Bolivia, in 2008, the under-five mortality rate is 54 per 1000 live births. Diarrhea causes 15% of these deaths, and 40% of pediatric diarrhea related hospitalizations are caused by rotavirus illness (RI). Rotavirus vaccination (RV), subsidized by international donors, is expected to reduce morbidity, mortality, and economic burden to the Bolivian state. Estimates of illness and economic burden of RI and their reduction by RV are essential to the Bolivian state's policies on RV program financing. The goal of this report is to estimate the economic burden of RI and the cost-effectiveness of the RV program. METHODS: To assess treatment costs incurred by the healthcare system, we abstracted medical records from 287 inpatients and 6751 outpatients with acute diarrhea between 2005 and 2006 at 5 sentinel hospitals in 4 geographic regions. RI prevalence rates were estimated from 4 years of national hospital surveillance. We used a decision-analytic model to assess the potential cost effectiveness of universal RV in Bolivia. RESULTS: Our model estimates that, in a 5-year birth cohort, Bolivia will incur over US$3 million in direct medical costs due to RI. RV reduces, by at least 60%, outpatient visits, hospitalizations, deaths, and total direct medical costs associated with rotavirus diarrhea. Further, RV was cost-savings below a price of US$3.81 per dose and cost-effective below a price of US$194.10 per dose. Diarrheal mortality and hospitalization inputs were the most important drivers of rotavirus vaccine cost-effectiveness. DISCUSSION: Our data will guide Bolivia's funding allocation for RV as international subsidies change. PMID- 21624422 TI - A protein-based pneumococcal vaccine protects rhesus macaques from pneumonia after experimental infection with Streptococcus pneumoniae. AB - Infections caused by Streptococcus pneumoniae are a major cause of mortality throughout the world. Protein-based pneumococcal vaccines are envisaged to replace or complement the current polysaccharide-based vaccines. In this context, detoxified pneumolysin (dPly) and pneumococcal histidine triad protein D (PhtD) are two potential candidates for incorporation into pneumococcal vaccines. In this study, the protective efficacy of a PhtD-dPly vaccine was evaluated in a rhesus macaque (Macaca mulatta) model of pneumonia. The animals were immunized twice with 10 MUg of PhtD and 10 MUg of dPly formulated in the Adjuvant System AS02 or with AS02 alone, before they were challenged with a 19F pneumococcal strain. The survival was significantly higher in the protein-vaccinated group and seemed to be linked to the capacity to greatly reduce bacterial load within the first week post-challenge. Vaccination elicited high concentrations of anti-PhtD and anti-Ply antibodies and a link was found between survival and antibody levels. In conclusion, AS02-adjuvanted PhtD-dPly vaccine protects against S. pneumoniae-induced pneumonia. It is probable that the protection is at least partially mediated by PhtD- and Ply-specific antibodies. PMID- 21624423 TI - Response of MUTZ-3 dendritic cells to the different components of the Haemophilus influenzae type B conjugate vaccine: towards an in vitro assay for vaccine immunogenicity. AB - Potency testing is mandatory for vaccine registration and batch release. Due to various limitations to in vivo potency testing, there is need for relevant in vitro alternatives. These alternative tests should preferably comprise cells from the target (human) species. The whole suite of immune responses to vaccination that occur in vivo in humans cannot be tested in vitro using a single cell type. Even so, dendritic cells (DC) form an important candidate cell type since they are pivotal in inducing and orchestrating immune responses. Cell lines are preferred over ex vivo cells for reasons of safety, accessibility, and reproducibility. In this first feasibility study we used the human cell line MUTZ 3, because it most closely resembles ex vivo human DC, and compared its response to monocyte-derived DC (moDC). Haemophilus influenzae type B (HiB) vaccine was chosen because its components exert different effects in vivo: while the HiB antigen, polyribosyl ribitol phosphate (PRP) fails to induce sufficient protection in children below 2 years of age, conjugation of this polysaccharide antigen to outer membrane protein (OMP) of Neisseria meningitides, results in sufficient protection. Effects of PRP, OMP, conjugated PRP-OMP, and adjuvanted vaccine (PedVax HiB), on cytokine production and surface marker expression were established. PRP induced no effects on cytokine production and the effect on surface marker expression was limited to a minor decrease in CD209 (DC-SIGN). In both MUTZ-3 and moDC, OMP induced the strongest response both in cytokine production and surface marker expression. Compared to OMP alone conjugated PRP OMP generally induced a weaker response in cytokine production and surface marker expression. The effects of PedVax HiB were comparable to conjugated PRP-OMP. While moDC showed a larger dynamic range than MUTZ-3 DC, these cells also showed considerable variability between donors, with MUTZ-3 DC showing a consistent response between the replicate assays. In our view, this makes MUTZ-3 DC the cells of choice. In conclusion, our results demonstrate that the MUTZ-3 DC assay allows discrimination between compounds with different immunogenicity. The potential of this cell line as (part of) an in vitro immunogenicity assay should be further explored. PMID- 21624424 TI - A meta-analysis of whole-brain diffusion tensor imaging studies in bipolar disorder. AB - OBJECTIVES: White matter abnormalities are one of the most consistently reported findings in neuroimaging studies of bipolar disorder (BD). We conducted an anatomical likelihood estimation meta-analysis of BD whole-brain diffusion tensor imaging (DTI) studies, with the aim of identifying statistically consistent fractional anisotropy (FA) changes reflecting microstructural modifications to white matter in BD. METHODS: We performed online searches of the PUBMED and EMBASE databases in January 2011. Studies were considered for inclusion if they used diffusion tensor MRI, compared a group of subjects with BD with healthy controls and involved whole-brain white matter analysis of FA. The analyses were conducted in Talairach space, using the activation likelihood estimation technique. We carried out a meta-analysis restricted to studies reporting a lower FA in patients with BD than in healthy controls. RESULTS: Ten studies were included. We identified two significant clusters of decreased FA on the right side of the brain. The first was located in the right white matter, close to the parahippocampal gyrus. Four of the ten studies included contributed to this cluster. The second cluster was located close to the right anterior and subgenual cingulate cortex. These two clusters of decreased FA in BD are crossed by several white matter tracts. CONCLUSIONS: These two clusters of altered FA may underlie the abnormal emotional processing and altered functional limbic connectivity in BD. Explorations based on DTI-based tractography are required to identify the tracts involved in the pathophysiology of BD. PMID- 21624425 TI - Neonatal hypothyroidism caused by maternal nicotine exposure is reversed by higher T3 transfer by milk after nicotine withdraw. AB - Maternal nicotine exposure leads to neonatal hypothyroidism that can be returned to euthyroidism after nicotine withdrawal. Here, we examined the transfer of iodine through milk, deiodinase activities (D1 and D2), and serum T3, T4 and TSH in rat offspring after maternal exposure to nicotine. One day after birth, a minipump was implanted to dams releasing nicotine (NIC), 6 mg/kg/day for 13 days or vehicle saline. Animals were killed at the day 15 and 21 of lactation. At day 15, NIC-treated dams showed decreased T4 and mammary 2h-radioiodine uptake (RAIU) and increase of TSH, thyroid 2h-RAIU, liver D1 and mammary D2. At the cessation of NIC-exposure, pups displayed decreased T3, T4 and thyroid 2h-RAIU and increased TSH. At weaning (21-postnatal day), NIC-treated dams recovered their T4 and TSH, but increased deiodinase level in the liver and mammary gland. Milk T3 content in NIC-treated dams was higher at both day 15 and 21, and thyroid function was recovered at the day 21. Thus, thyroid function was affected by nicotine in both mothers and pups, suggesting a primary hypothyroidism. After nicotine withdrawal, pups recovered thyroid function probably due to the increased lactational transfer of T3 in relation with increased mammary gland deiodinase activities. PMID- 21624426 TI - Kasstasin: A novel potent vasoconstrictor peptide from the skin secretion of the African red-legged running frog, Kassina maculata. AB - Amphibian skin secretions are established sources of bioactive peptides. Here we describe the isolation, structural and pharmacological characterisation of a novel vasoconstrictor peptide from the skin secretion of the African hyperoliid frog, Kassina maculata, which exhibits no structural similarity to any known class of amphibian skin peptide. The peptide consists of 21 amino acid residues, FIKELLPHLSGIIDSVANAIK, and is C-terminally amidated. The provisional structure was obtained by MS/MS fragmentation using an Orbitrap mass spectrometer and L/I ambiguities were resolved following molecular cloning of biosynthetic precursor encoding cDNA. A synthetic replicate of the peptide was found to possess weak antimicrobial and haemolytic activities but was exceptionally effective in constricting the smooth muscle of rat tail artery (EC(50) of 25 pM). In reflection of its exceptional potency in constricting rat arterial smooth muscle, the peptide was named kasstasin, a derivation of Kassina and "stasis" (stoppage of flow). These data illustrate the continuing potential of amphibian skin secretions to provide novel natural peptide templates for biological evaluation. PMID- 21624427 TI - In and out of the niche: perspectives in mobilization of hematopoietic stem cells. AB - Several stem cell mobilization strategies have been employed in the past 2 decades, including chemotherapy, hematopoietic growth factors, and chemotherapy plus growth factors. Granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage CSF are standard agents approved for peripheral blood stem cell mobilization since the early 1990s. Between 5% and 20% of patients, however, fail to mobilize a sufficient numbers of peripheral blood stem cells in response to G-CSF with or without chemotherapy. Recent advances in defining the basic mechanisms regulating the interactions between hematopoietic stem cells and their marrow niche had led to the discovery that CXCR4 and stromal-cell-derived factor 1alpha axis play a significant role. Plerixafor, an antagonist of the CXCR4 stromal-cell-derived factor 1alpha axis has been shown to result in a significant mobilization of hematopoietic stem cells. Numerous clinical trials have demonstrated that the combination of G-CSF and AMD3100 (G+A) resulted in a significant increase in CD34(+) cell yield as compared to the administration of G CSF alone. In particular, the progenitors mobilized have been shown to comprise a significantly higher proportion of primitive and possibly more potent CD34(+)/CD38(-) subpopulation. Transplantation of PBSC mobilized by G+A administration have led to a rapid and sustained neutrophil and platelet engraftment. Another prospective role of this new class of agents might lie in the mobilization of dormant leukemia stem cells that are well protected by the niche. The future role of CXCR4 antagonists in treatment of hematologic malignancies includes mobilization of hematopoietic stem cells for transplantation and mobilization of leukemia-initiating cells for long-term cure. PMID- 21624428 TI - Decrease of prestin expression by increased potassium concentration in organotypic cultures of the organ of Corti of newborn rats. AB - Prestin is the motor protein of the outer hair cells of the organ of Corti and a key factor in ensuring a high sensitivity level of mammalian hearing. In the present study, we examined the effects of increased extracellular potassium (K(+)) concentration on the expression of prestin mRNA and the transcription factors Gata-3 and Carf in the organotypic culture of the organ of Corti of newborn rats. Mannitol and NaCl were used to analyze possible effects of hyperosmotic stress or ion-specific changes, respectively. An increase in prestin expression by a factor of 1.5-2.0 was seen in cultures grown in the presence of 5mM K(+). Potassium concentration of 35 and 55 mM induced a parallel decrease in prestin and Carf expression, but Gata-3 expression increased. Mannitol had no effect on gene expression whereas increased NaCl concentrations decreased prestin, but not Carf expression. The data suggest that chronic depolarization might decrease the prestin expression and possibly contribute to hearing loss and tinnitus. PMID- 21624429 TI - Silencing of Nogo-A in rat oligodendrocyte cultures enhances process branching. AB - The myelin-associated protein Nogo-A is a well-known inhibitor for axonal regeneration and compensatory plasticity, yet functions of endogenous Nogo-A in oligodendrocyte differentiation are not as clear. As oligodendrocyte matures, its processes branch and eventually form lamellae that ensheath target axons. The present study examined the effects of decreased levels of Nogo-A on the development of oligodendrocytes. The siRNA mediated Nogo-A silencing in these cells did not change their proliferation rates identified by BrdU incorporation assay and neither the expression of stage specific oligodendrocyte makers as identified by qRT-PCR and immunostaining. But knockdown the expression of Nogo-A significantly enhances the process branching complexity by Sholl analysis. Current results suggest a novel role for Nogo-A in maintaining a restricted branching phenotype in oligodendrocytes process outgrowth, which is a key step towards myelin membrane sheet formation and myelination. PMID- 21624430 TI - Connectivity brain networks based on wavelet correlation analysis in Parkinson fMRI data. AB - Recent studies have shown that aging, psychiatric and neurologic diseases, and dopaminergic blockade all result in altered brain network efficiency. We investigated the efficiency of human brain functional networks as measured by fMRI in individuals with idiopathic Parkinson's disease (N=14) compared to healthy age-matched controls (N=15). Functional connectivity between 116 cortical and subcortical regions was estimated by wavelet correlation analysis in the frequency interval of 0.06-0.12 Hz. Efficiency of the associated network was analyzed, comparing PD to healthy controls. We found that individuals with Parkinson's disease had a marked decrease in nodal and global efficiency compared to healthy age-matched controls. Our results suggest that algorithmic approach and graph metrics might be used to identify and track neurodegenerative diseases, however more studies will be needed to evaluate utility of this type of analysis for different disease states. PMID- 21624431 TI - Muscarinic receptor immunoreactivity in the superior salivatory nucleus neurons innervating the salivary glands of the rat. AB - The superior salivatory nucleus (SSN) contains preganglionic parasympathetic neurons to the submandibular and sublingual salivary glands. Cevimeline, a muscarinic acetylcholine receptor agonist, stimulates the salivary glands and is presently used as sialogogue in the treatment of dry mouth. Since cevimeline passes through the blood-brain barrier, it is also able to act on muscarinic acetylcholine receptors in the central nervous system. Our preliminary experiment using the whole-cell patch-clamp technique has shown that cevimeline excites SSN neurons in rat brain slices, suggesting that SSN neurons have muscarinic acetylcholine receptors; however, it is unclear which subtypes of muscarinic acetylcholine receptors exist in SSN neurons. In the present study, we investigated immunohistochemically muscarinic acetylcholine receptor subtypes, M1 receptor (M1R), M2R, M3R, M4R, and M5R in SSN neurons. SSN neurons innervating the salivary glands, retrogradely labeled with a fluorescent tracer from the chorda-lingual nerve, mostly expressed M3R immunoreactivity (-ir) (92.3%) but not M1R-ir. About half of such SSN neurons also showed M2R- (40.1%), M4R- (54.0%) and M5R-ir (46.0%); therefore, it is probable that SSN neurons co-express M3R-ir with at least two of the other muscarinic receptor subtypes. This is the first report to show that SSN neurons contain muscarinic acetylcholine receptors. PMID- 21624432 TI - Post-learning REM sleep deprivation impairs long-term memory: reversal by acute nicotine treatment. AB - Rapid eye movement sleep deprivation (REM-SD) is associated with spatial learning and memory impairment. During REM-SD, an increase in nicotine consumption among habitual smokers and initiation of tobacco use by non-smokers have been reported. We have shown recently that nicotine treatment prevented learning and memory impairments associated with REM-SD. We now report the interactive effects of post learning REM-SD and/or nicotine. The animals were first trained on the radial arm water maze (RAWM) task, then they were REM-sleep deprived using the modified multiple platform paradigm for 24h. During REM-SD period, the rats were injected with saline or nicotine (1mg/kg s.c. every 12h: a total of 3 injections). The animals were tested for long-term memory in the RAWM at the end of the REM-SD period. The 24h post-learning REM-SD significantly impaired long-term memory. However, nicotine treatment reversed the post-learning REM-SD-induced impairment of long-term memory. On the other hand, post-learning treatment of normal rats with nicotine for 24h enhanced long-term memory. These results indicate that post learning acute nicotine treatment prevented the deleterious effect of REM-SD on cognitive abilities. PMID- 21624433 TI - Experience with novel actions modulates frontal alpha EEG desynchronization. AB - There is growing interest in the effects of experience on the neural processes linking action execution and action perception. We tested whether experience with unfamiliar actions can alter desynchronization of alpha-range power in the EEG upon re-observation of those actions. In a training session, participants (N=21) watched videos of novel drawing movements. Half of the movements were imitated after each viewing, and half of the movements were seen but not imitated, thus forming two training conditions: visual plus motor experience (V+M), and visual experience only (VO). In a testing session the next day, participants were shown the same videos of both sets of movements, and were also shown a third, completely novel, set of movements. Imitative performance was better for both training conditions than for novel actions. Event-related EEG desynchronization in the upper alpha band during action observation differed between conditions at frontal electrode sites, with novel actions being associated with less frontal desynchronization compared to V+M and VO actions. Differences between conditions were not noted over other regions. This suggests that moderate amounts of initial experience with novel actions can alter the neural processing of these actions when viewed again, particularly over frontal regions. PMID- 21624434 TI - Memory of fearful events: the role of fibroblast growth factor-2 in fear acquisition and extinction. AB - Research during the past decade has led to a tremendous growth in our understanding of how fear memories are acquired and subsequently inhibited on a neural and molecular level. Such research has contributed to significant developments in the treatment of anxiety disorders, and has considerably advanced our understanding of the neurobiology of learning and memory in general. A number of recent studies have examined the role of growth factors in the formation of long-term memory for fearful events, due to their ability to cause morphological neural changes in response to environmental stimulation. In this review we first describe physiological evidence that fibroblast growth factor-2 (FGF2) receptors are highly expressed in the neural circuitry regulating fear acquisition and extinction, and that FGF2 modulates the molecular signals known to be involved in the formation of fear memories. Then we present emerging behavioral research that demonstrates that exogenous FGF2 can enhance the formation of fear conditioning and extinction memories. Finally, we briefly discuss how research into the role of FGF2 in learning and memory may be of clinical benefit, particularly in the treatment of anxiety disorders. PMID- 21624436 TI - The role of oxidative stress in neuromelanin synthesis in PC12 cells. AB - Previous research has indicated that neuromelanin (NM) is involved in the pathogenesis of Parkinson's disease (PD). Increased reactive oxygen species (ROS) generation in PD sufferers is thought to be related to enhanced tyrosine hydroxylase (TH) activity and NM production. However, few reports have confirmed this hypothesis. In this study, PC12 cells of all experiments were exposed to 50 MUmol/L levodopa (l-DOPA) to generate a model for NM synthesis. Meanwhile, PC12 cells were treated with glucose oxidase (GO) at different concentrations to generate oxidative stress. Finally, cell viability, TH activity, and NM generation in PC12 cells were measured. The results showed that GO dose dependently stimulated oxidative stress generation in PC12 cells. Moderate increases in oxidative stress enhanced the viability of PC12 cells. However, an excessive level of oxidative stress can lead to the degeneration of PC12 cells. Notably, in the surviving PC12 cells, ROS significantly increased the TH activity, and the NM production was also upregulated. Thus, oxidative stress may upregulate the synthesis of NM, which may be a result of the increased TH activity observed in response to the elevated ROS in l-DOPA-treated PC12 cells. PMID- 21624435 TI - Neuroanatomic and behavioral traits for autistic disorders in age-specific restricted index selection mice. AB - The pathogenesis of neurodevelopmental disorders such as autism is believed to be influenced by interactions between genetic and environmental factors, and appropriate animal models are needed to assess the influence of such factors on relevant neurodevelopmental phenotypes. A set of inbred mouse strains (Atchley strains) including A12 (E+L0) and A22 (E-L0) were generated by age-specific restricted index selection from a baseline random-bred ICR mouse population obtained from Harlan Sprague-Dawley [Atchley et al. (1997) Genetics 146(2):629 640; Indianapolis, IN, USA). As compared with the A22 strain, A12 mice had significantly increased early (P0-P10) body weight gain with minimal changes in late (P28-P56) body weight gain. We found that these strains also differed in brain weight, brain volume, cell proliferation, and FGF-2 levels in certain brain regions. Specifically, brain weight and volume were significantly greater in A12 mice than that in A22 mice at P10 and P28. Quantitative analysis of bromodeoxyuridine (BrdU) labeling of proliferating cells showed that the number of BrdU-positive cells in the A12 strain were significantly greater in the frontal cortex and lesser in the dentate gyrus than that in the A22 strain at P28. Western blot revealed that fibroblast growth factors-2 (FGF-2), but not brain-derived neurotrophic factor (BDNF), expression was significantly increased in the frontal cortex of A12 strain at P28. Also, A12 mice exhibited decreased intra-strain social interaction and increased repetitive stereotyped behaviors at P28. Our study suggests that A12 mice may partially mimic the anatomic and behavioral traits of patients with neurodevelopmental disorders such as autism spectrum disorders, and therefore may yield insights into the developmental mechanisms involved in their pathogenesis. PMID- 21624437 TI - What the nose knows, what the eyes see, how we feel, how we learn, how we understand motor acts, why "YY" is essential for ion transport, how epigenetics meet neurobiology in Rett syndrome: seven topics at the 2010 Kavli Prize Symposium on Neuroscience. PMID- 21624438 TI - F-actin depolymerization accelerates clasmatodendrosis via activation of lysosome derived autophagic astroglial death. AB - Clasmatodendrosis is an irreversible astroglial degenerative change, which includes extensive swelling and vacuolization of cell bodies and disintegrated and beaded processes. Since alteration in F-actin level influences on the formation of vacuoles/vesicles during exocytosis/endocytosis in astrocytes, we investigated whether F-actin polymerization involves clasmatodendrosis in the rat hippocampus following status epilepticus (SE). In the present study, vacuoles in clasmatodendrotic astrocytes showed LAMP-1 and LC3-II (a marker for autophagy) immunoreactivity. These findings reveal that clasmatodendrosis may be lysosome derived autophagic astroglial death. Jasplakinolide (an F-actin stabilizer) infusion significantly decreased the size and the number of medium/large-sized vacuoles in each clasmatodendritic astrocyte accompanied by enhancement of phalloidin signals, as compared to vehicle-infusion. In contrast, latrunculin A (an F-actin-depolymerizing agent) infusion increased the size and the number of medium/large-sized vacuoles, which were dissociated adjacent to cell membrane. Therefore, our findings suggest that F-actin stabilization may inhibit lysosome derived autophagic astroglial death during clasmatodendrosis. PMID- 21624439 TI - Brief neck restraint stress enhances long-term potentiation and suppresses long term depression in the dentate gyrus of the mouse. AB - We studied the effects of brief (10 min) neck restraint on long-term potentiation (LTP) and long-term depression (LTD) in mouse dentate gyrus (DG) slices. Brain slices were prepared immediately after neck restraint and LTP/LTD induction was attempted 3.5h later. LTP enhancement and LTD suppression was observed in slices prepared from stressed animals. The corticosterone plasma concentration was elevated approx. fourfold just after the neck restraint session. To examine the role of glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) activation in mediating the effects of neck restraint on LTP and LTD, the animals were pretreated with either the specific GR antagonist RU38486, the specific MR antagonist spironolactone, the corticosterone synthesis inhibitor metyrapone, or vehicle Tween 80. Injection with Tween 80 and subsequent neck restraint did not influence LTP enhancement or LTD suppression, but the effects of neck restraint were prevented by RU38486, spironolactone and metyrapone pretreatment. These data indicate that acute stress caused by brief neck restraint enhances LTP and impairs LTD in the dentate gyrus and that these effects are mediated by activation of GRs and MRs. PMID- 21624440 TI - Alterations of emotion, cognition and firing activity of the basolateral nucleus of the amygdala after partial bilateral lesions of the nigrostriatal pathway in rats. AB - Although increasing evidence indicates that psychiatric symptoms are crucial characteristic of the early stage of Parkinson's disease (PD) and precede motor impairments, the neuronal firing activity of the basolateral nucleus of the amygdala (BLA) in the psychiatric symptom of PD and the involved mechanism are still unclear. In the present study, we examined the changes in emotional and cognitive tests not focused on motor fluency and firing activity of projection neurons in the BLA rats with 6-hydroxydopamine (6-OHDA) injected bilaterally into dorsal striatum, and the effects of apomorphine and the medial prefrontal cortex (mPFC) on these changes. Injection of 6-OHDA (10.5 MUg) into the dorsal striatum produced 18-22% and 26-30% loss of tyrosine hydroxylase immunoreactive neurons in the ventral tegmental area and substantia nigra pars compacta of rats, respectively. The striatal lesions induced anxiety-like responses in the rats but did not result in depressive-like behavior or cognitive impairments. In the lesioned rats, the firing rate of BLA projection neurons decreased significantly compared with sham-operated rats, and the firing pattern of BLA projection neurons was not changed. No significant differences were observed either in behaviors or firing activity of BLA projection neurons by further ibotenic acid lesions of the mPFC in the lesioned rats. Systemic administration of cumulative apomorphine (10-160 MUg/kg) inhibited the firing rate of BLA projection neurons in sham-operated, 6-OHDA-lesioned and combined 6-OHDA- and mPFC-lesioned rats, but the latter needed more apomorphine stimulation. These data suggest that the anxiety in early stage of PD is possibly related to the decrease in firing activity of BLA projection neurons, which may be regulated by the activation of dopamine receptor in the mPFC. PMID- 21624441 TI - Selective cognitive impairment in the YAC128 Huntington's disease mouse. AB - People with HD have a demonstrated early extra-dimensional set-shifting deficit. In the present study, we use a novel water T-maze set-shifting procedure and demonstrate its validity as a set-shifting task in a mouse model of Huntington's disease. Three groups of YAC128 mice of different ages (27, 69 and 117 weeks) were run on the task, which incorporated six distinct stages in which the mice must learn a rule and then switch to a different rule. The six stages were: directional learning, directional learning reversal, light discrimination, light discrimination reversal, return to place learning and a maze rotation spatial learning test. Rule changes from place learning to light discrimination and back constitute extra-dimensional shifts. The results of the study demonstrate robust light/dark discrimination reversal learning deficits in transgenic mice from 27 weeks of age, and a directional learning to light discrimination extra dimensional set-shifting deficit from 69 weeks of age. The extra-dimensional shift deficit was confirmed with control trials demonstrating the validity of the deficit and the task. The onset of reversal learning and extra-dimensional shift deficits corresponded with the development of mutant huntingtin N-terminal fragment aggregates in neurons of relevant forebrain regions. PMID- 21624442 TI - Suppression of beta-naphthoflavone induced CYP1A expression and lipid peroxidation by berberine. AB - Impacts of berberine, a major isoquinoline alkaloid in herbal plants, on beta naphthoflavone (BNF)-induced CYP1A expression were determined both in primary mouse hepatocytes and in vivo. Berberine concentration-dependently suppressed BNF induced CYP1A expression in mouse hepatocyte and it significantly down-regulated BNF-induced CYP1A in mouse liver via suppression of mRNA and protein expression, and decreases of EROD and MROD activities. Moreover, berberine showed significant potential on suppression of BNF-induced lipid peroxidation in mouse hepatic microsome. Therefore, using berberine as a health supplement or an alternative medication might provide extra-benefit due to its inhibitory regulation on CYP1A expression and anti-lipid peroxidation activity. PMID- 21624443 TI - Antitrypanosomal sesquiterpene lactones from Saussurea costus. AB - In the course of a larger screen of 1800 plant and fungal extracts, the ethyl acetate extract of Saussurea costus roots potently inhibited the growth of Trypanosoma brucei rhodesiense. Subsequent HPLC based activity profiling led to the identification of the sesquiterpene lactones arbusculin B (1), alpha cyclocostunolide (2), costunolide (3), and dehydrocostuslactone (4). They were tested for in vitro antitrypanosomal activities and cytotoxicity alongside the structurally related sesquiterpene lactones parthenolide (5), zaluzanin D (6), and eupatoriopicrin (7), and had IC(50)s between 0.8 and 22 MUM. Cytotoxic IC(50)s were from 1.6 to 19 MUM, and selectivity indices from 0.5 to 6.5. PMID- 21624444 TI - Oleanene triterpenes from Sedum lineare Thunb. AB - One new olean-13(18)-ene-3,12,19-trione (1), and two known oleanene triterpenes delta-amyrone (2), and delta-amyrine acetate (3) were isolated from the petroleum ether fraction from an alcoholic extract of the whole plant of Sedum linear Thunb. The new compound was characterized by means of spectroscopic methods including 1D, 2D NMR and HR-ESI-MS, and was further confirmed by X-ray diffraction analysis. The known ones were established on the basis of comparing their NMR data with those of the corresponding compounds in the literature. In addition, the inhibitory effects of the compounds isolated on the TNF-alpha and NO production were examined in vitro. PMID- 21624445 TI - Acute inhalation toxicity of cerium oxide nanoparticles in rats. AB - The aim of the present study was to assess the acute toxic potential of cerium oxide nanoparticles (CeO(2) NPs) in rats when exposed through the head and nose inhalation route. The rats were exposed to CeO(2) NPs and the resultant effects if any, to cause cytotoxicity, oxidative stress and inflammation in the lungs were evaluated on a 24h, 48h and 14 day post exposure period. Our results showed a significant decrease in the cell viability, with the increase of lactate dehydogenase, total protein and alkaline phosphatase levels in the bronchoalveolar lavage fluid (BALF) of the exposed rats. Total leukocyte count and the percentage of neutrophils in BALF were elevated within 24h of post exposure. The concentrations of pro-inflammatory cytokines (IL-1beta, TNF-alpha, and IL-6) were significantly increased in the BALF and in the blood throughout the observation period. The level of malondialdehyde was elevated with the decreased levels of intracellular reduced glutathione (GSH) in the lung after exposure. The alveolar macrophages (AMs) and neutrophils overloaded with phagocytosed CeO(2) NPs were observed along with non-phagocytosed free CeO(2) NPs that were deposited over the epithelial surfaces of the bronchi, bronchiole and alveolar regions of lungs within 24h of post exposure and were consistent throughout the observation period. A well distributed, multifocal pulmonary microgranulomas due to impairment of clearance mechanism leading to biopersistence of CeO(2) NPs for an extended period of time were observed at the end of the 14 day post exposure period. These results suggest that acute exposure of CeO(2) NPs through inhalation route may induce cytotoxicity via oxidative stress and may lead to a chronic inflammatory response. PMID- 21624446 TI - The study on venom proteins of Lapemis hardwickii by cDNA phage display. AB - In this study, a cDNA T7 phage display library was constructed from sea snake Lapemis hardwickii venom gland mRNA to analyze the components in venom and find new toxins. All the venom gland cDNA-encoding proteins, including housekeeping proteins and venom proteins were expressed on the surface of bacteriophage T7. This library was then panned with rabbit anti-sea snake venom IgG. Phage particles displaying venom-components with interaction with the antibodies were enriched. Thus, phage-carrying venom proteins, such as short chain neurotoxin, long chain neurotoxin, PLA2-like toxin, and c-type lectin-like protein were found, some of them were never reported previously. Four different short chain neurotoxins (SNT-1, 2, 3, 4) were cloned and expressed in Escherichia coli. The LD(50) and analgesic activities of their purified forms were determined. Their structure-function relationship were studied with the aid of a toxin-nAChR complex model constructed by ourselves. Among them, SNT-4 was a new neurotoxin identified in this study and showed potential as pain killer. These results prove that cDNA phage display technique has great advantage than traditional cDNA library method because of the linkage between phenotype and genotype of phage, which provided an effective means to study unknown proteins with target functions. PMID- 21624447 TI - Drawdown of floating solids in stirred tanks: scale-up study using CFD modeling. AB - This work shows development of a scale up correlation using computational fluid dynamic (CFD) simulations for floating solids drawdown operation in stirred tanks. Discrete phase modeling (DPM) simulations were used in conjunction with the lab scale experimental measurements to develop a semi-empirical correlation for the prediction of rate of drawdown of floating solid particles. The rate was correlated to average liquid velocity at the free liquid surface. Since, this correlation is based on a fundamental hydrodynamic parameter, velocity, rather than an operating parameters such as the impeller speed, it can be used for a variety of impeller types and tank geometries. The correlation was developed based on the data obtained from the 2L tank using four different tank designs and was validated against the data obtained from the 10L scale tank. The correlation was further extended to the pilot and the commercial scale tanks ranging from 40L to 4000L scale based solely on the CFD model. PMID- 21624448 TI - Radial tensile strength prediction of relaxing and relaxed compacts by near infrared chemical imaging. AB - The prediction of radial tensile strength (RTS) of relaxing solid dosage forms by near-infrared hyperspectral chemical imaging was studied. Compacts consisting of starch, lactose, and a mixture of four components were created at different compression forces to develop density models. Predicted density distribution parameters were subsequently used to estimate RTS. Chemical images were collected shortly after compression, repeated every 30 min for 2h, and a final image was collected after 2 weeks. A two step process, involving first the prediction of compact density at each pixel (using a partial least squares model) and second the relationship between compact density distributions and RTS was implemented. Among the parameters with a significant relationship with RTS, the median of the distribution of density predictions in an image was identified as a robust parameter. Coefficients of determinations for this prediction ranged from 0.96 to 0.99 were obtained with a maximum error in validation of 0.10 MPa for the four component formulation compacts. The prediction of RTS of fully relaxed compacts from spectral data collected on relaxing compacts was demonstrated. These results demonstrate the potential to use near-infrared chemical imaging in real-time to predict RTS values of fully relaxed compacts. PMID- 21624449 TI - Understanding deformation mechanisms during powder compaction using principal component analysis of compression data. AB - Principal component analysis (PCA) was applied to pharmaceutical powder compaction. A solid fraction parameter (SF(c/d)) and a mechanical work parameter (W(c/d)) representing irreversible compression behavior were determined as functions of applied load. Multivariate analysis of the compression data was carried out using PCA. The first principal component (PC1) showed loadings for the solid fraction and work values that agreed with changes in the relative significance of plastic deformation to consolidation at different pressures. The PC1 scores showed the same rank order as the relative plasticity ranking derived from the literature for common pharmaceutical materials. The utility of PC1 in understanding deformation was extended to binary mixtures using a subset of the original materials. Combinations of brittle and plastic materials were characterized using the PCA method. The relationships between PC1 scores and the weight fractions of the mixtures were typically linear showing ideal mixing in their deformation behaviors. The mixture consisting of two plastic materials was the only combination to show a consistent positive deviation from ideality. The application of PCA to solid fraction and mechanical work data appears to be an effective means of predicting deformation behavior during compaction of simple powder mixtures. PMID- 21624450 TI - Forward and reverse transfer functions of the middle ear based on pressure and velocity DPOAEs with implications for differential hearing diagnosis. AB - Recently it was shown that distortion product otoacoustic emissions (DPOAEs) can be measured as vibration of the human tympanic membrane in vivo, and proposed to use these vibration DPOAEs to support a differential diagnosis of middle-ear and cochlear pathologies. Here, we investigate how the reverse transfer function (r TF), defined as the ratio of DPOAE-velocity of the umbo to DPOAE-pressure in the ear canal, can be used to diagnose the state of the middle ear. Anaesthetized guinea pigs served as the experimental animal. Sound was delivered free-field and the vibration of the umbo measured with a laser Doppler vibrometer (LDV). Sound pressure was measured 2-3 mm from the tympanic membrane with a probe-tube microphone. The forward transfer function (f-TF) of umbo velocity relative to ear canal pressure was obtained by stimulating with multi-tone pressure. The r-TF was assembled from DPOAE components generated in response to acoustic stimulation with two stimulus tones of frequencies f(1) and f(2); f(2)/f(1) was constant at 1.2. The r-TF was plotted as function of DPOAE frequencies; they ranged from 1.7 kHz to 23 kHz. The r-TF showed a characteristic shape with an anti-resonance around 8 kHz as its most salient feature. The data were interpreted with the aid of a middle-ear transmission-line model taken from the literature for the cat and adapted to the guinea pig. Parameters were estimated with a three-step fitting algorithm. Importantly, the r-TF is governed by only half of the 15 independent, free parameters of the model. The parameters estimated from the r-TF were used to estimate the other half of the parameters from the f-TF. The use of r-TF data - in addition to f-TF data - allowed robust estimates of the middle-ear parameters to be obtained. The results highlight the potential of using vibration DPOAEs for ascertaining the functionality of the middle ear and, therefore, for supporting a differential diagnosis of middle-ear and cochlear pathologies. PMID- 21624451 TI - Biomass pretreatment: fundamentals toward application. AB - Development of sustainable energy systems based on renewable biomass feedstocks is now a global effort. Lignocellulosic biomass contains polymers of cellulose, hemicellulose, and lignin, bound together in a complex structure. Liquid biofuels, such as ethanol, can be made from biomass via fermentation of sugars derived from the cellulose and hemicellulose within lignocellulosic materials, but the biomass must be subjected to pretreatment processes to liberate the sugars needed for fermentation. Production of value-added co-products along-side biofuels through integrated biorefinery processes creates the need for selectivity during pretreatment. This paper presents a survey of biomass pretreatment technologies with emphasis on concepts, mechanism of action and practicability. The advantages and disadvantages, and the potential for industrial applications of different pretreatment technologies are the highlights of this paper. PMID- 21624452 TI - Identification and characterization of wblA-dependent tmcT regulation during tautomycetin biosynthesis in Streptomyces sp. CK4412. AB - Tautomycetin (TMC) is an unusual linear polyketide compound esterified with a cyclic anhydride. It exhibits novel activated T cell-specific immunosuppressant as well as anti-cancer activities. Previously, we isolated and characterized the entire TMC biosynthetic gene cluster from Streptomyces sp. CK4412, including a TMC pathway-specific gene, tmcN, the over-expression of which led to a significant increase in TMC productivity. In addition, we also reported that WblA acts as a global down-regulator of antibiotic biosynthesis through pathway specific regulation in Streptomyces species. Here, we confirm that TmcT acts as another TMC pathway-specific regulator within the TMC biosynthetic cluster. Specifically, tmcT deletion resulted in the complete loss of TMC production, whereas complementation with a tmcT-carrying integrative plasmid significantly restored TMC biosynthesis. We also identified a 0.39kb wblA ortholog (named wblA(tmc)) from Streptomyces sp. CK4412 via genomic DNA library screening that showed 96% amino acid identity compared to a previously-known S. coelicolor wblA. Targeted gene disruption of wblA(tmc) in Streptomyces sp. CK4412 exhibited approximately 3-fold higher TMC productivity than that in the wild-type strain. Moreover, transcription analyses of the TMC biosynthetic and regulatory genes revealed that the expression of tmcT was strongly down-regulated by wblA(tmc). These results imply that the TMC biosynthetic regulation network is controlled by two pathway-specific positive regulator, WblA(tmc)-dependent TmcT as well as WblA(tmc)-independent TmcN in Streptomyces sp. CK4412. PMID- 21624453 TI - Harnessing the power of enzymes for environmental stewardship. AB - Enzymes are versatile catalysts with a growing number of applications in biotechnology. Their properties render them also attractive for waste/pollutant treatment processes and their use might be advantageous over conventional treatments. This review highlights enzymes that are suitable for waste treatment, with a focus on cell-free applications or processes with extracellular and immobilized enzymes. Biological wastes are treated with hydrolases, primarily to degrade biological polymers in a pre-treatment step. Oxidoreductases and lyases are used to biotransform specific pollutants of various nature. Examples from pulp and paper, textile, food and beverage as well as water and chemical industries illustrate the state of the art of enzymatic pollution treatment. Research directions in enzyme technology and their importance for future development in environmental biotechnology are elaborated. Beside biological and biochemical approaches, i.e. enzyme prospection and the design of enzymes, the review also covers efforts in adjacent research fields such as insolubilization of enzymes, reactor design and the use of additives. The effectiveness of enzymatic processes, especially when combined with established technologies, is evident. However, only a limited number of enzymatic field applications exist. Factors like cost and stability of biocatalysts need to be addressed and the collaboration and exchange between academia and industry should be further strengthened to achieve the goal of sustainability. PMID- 21624454 TI - The clinically available NMDA receptor antagonist, memantine, exhibits relative safety in the developing rat brain. AB - The N-methyl-d-aspartate glutamate receptor (NMDAR) has been implicated in preterm brain injury (periventricular leukomalacia (PVL)) and represents a potential therapeutic target. However, the antagonist dizocilpine (MK-801) has been reported to increase constitutive neuronal apoptosis in the developing rat brain, limiting its clinical use in the developing brain. Memantine is another use-dependent NMDAR antagonist with shorter binding kinetics and has been demonstrated to be protective in a rat model of PVL, without effects on normal myelination or cortical growth. To further evaluate the safety of memantine in the developing brain, we demonstrate here that, in contrast to MK-801, memantine at neuroprotective doses does not increase neuronal constitutive apoptosis. In addition, there are no long-term alterations in the expression of NMDAR subunits, AMPAR subunits, and two markers of synaptogenesis, Synapsin-1 and PSD95. Evaluating clinically approved drugs in preclinical neonatal animal models of early brain development is an important prerequisite to considering them for clinical trial in preterm infants and early childhood. PMID- 21624455 TI - A discussion of the impact of US chemical regulation legislation on the field of toxicity testing. AB - Proposals for revising the principal United States law governing industrial chemicals, the Toxic Substances Control Act, have been under consideration in the US Congress for the past several years, and some version of such legislation may be passed in the near future. Concurrently, a desire to move away from current testing methods for ethical, scientific, and practical reasons has led to multi million dollar investments in in vitro and computational toxicology methods and programs. Legislative language has the potential to endorse this transition and facilitate its fruition, or conversely enshrine in vivo testing methods for the foreseeable future. New legislation also has the potential to substantially increase the numbers of animals used in toxicity tests in the near term. However, there are a number of policies that, used effectively, can reduce the overall number of animals used in new toxicity tests. We present recent legislative proposals in the context of current testing programs and discuss their potential impacts on animal use, test method innovation, and achievement of desired legislative objectives. Discussions like these are essential to judiciously select policies that reduce the use of animals in toxicity testing and protect human health and the environment. PMID- 21624456 TI - Genistein impairs early testosterone production in fetal mouse testis via estrogen receptor alpha. AB - The widespread consumption of soy-based products raises the issue of the reproductive toxicity of phytoestrogens. Indeed, it is well known that genistein, an isoflavone found in soybeans and soy products, mimics the actions of estrogens and that the fetal testis is responsive to estrogens. Therefore we investigated whether genistein could have deleterious effects on fetal testis. Using organ cultures of fetal testes from wild type and ERalpha or ERbeta knock-out mice we show that genistein inhibits testosterone secretion by fetal Leydig cells during early fetal development (E12.5), within the "masculinization programming window". This effect occurs through an ERalpha-dependent mechanism and starting at 10 nM genistein, a concentration which is compatible with human consumption. No effect of genistein on the number of gonocytes was detected at any of the studied developmental stages. These results suggest that fetal exposure to phytoestrogens can affect the development and function of the male reproductive system. PMID- 21624457 TI - Looking ultra deep: short identical sequences and transcriptional slippage. AB - Studying transcriptomes by ultra deep sequencing provides an in-depth picture of transcriptional regulation and it facilitates the detection of rare transcriptional events. Using ultra deep sequencing of amplicons we identified known isoforms and also various new low frequency variants. Most of these variants likely involve the splicing machinery except for two events that we named variations affecting multiple exons, which are mainly deletions affecting parts of adjacent exons and intra-exonic deletions. Both events involve short identical sequences of 1 to 8 nucleotides at the junction and canonical splice sites are missing. They were identified in different genes and species at very low frequencies. We excluded that they are an artifact of PCR, sequencing, or reverse transcription. We propose that these variants represent intramolecular slippage events that require short identical sequences for reannealing of dissociated transcripts. PMID- 21624458 TI - The Trypanosoma cruzi genome; conserved core genes and extremely variable surface molecule families. AB - The protozoan parasite Trypanosoma cruzi is an important but neglected pathogen that causes chagas disease, which affects millions of people, mainly in latin America. The population structure and epidemiology of the parasite are complex, with much variability among strains. The genome sequence of a reference strain, CL Brener, was published in 2005, and the availability of this sequence has both revealed the complexity of the parasite genome and greatly facilitated research into parasite biology and pathogenesis, by making the sequences of more than 8000 core genes available. The T. cruzi genome is highly repetitive, which has resulted in inaccuracies in the genome sequence, and attempts have been made to provide a deeper analysis of repeated genes as a complement to the genome sequence. The genome was found to be organized in stable core regions containing housekeeping and other genes, surrounded by highly repetitive, often sub telomeric highly variable regions containing multiple members of large families of surface molecule genes. Comparative sequencing of T. cruzi strains has been initiated and the results show that the core gene content of the parasite is highly conserved, but that much sequence variability, 3-4% difference at the DNA level on average between strains in coding regions, is present. The additional genomes will improve the understanding of parasite biology and epidemiology. PMID- 21624459 TI - Tetrahymena thermophila, a unicellular eukaryote with separate germline and somatic genomes. AB - Tetrahymena thermophila is a ciliate--a unicellular eukaryote. Remarkably, every cell maintains differentiated germline and somatic genomes: one silent, the other expressed. Moreover, the two genomes undergo diverse processes, some as extreme as life and death, simultaneously in the same cytoplasm. Conserved eukaryotic mechanisms have been modified in ciliates to selectively deal with the two genomes. We describe research in several areas of Tetrahymena biology, including meiosis, amitosis, genetic assortment, selective nuclear pore transport, somatic RNAi-guided heterochromatin formation, DNA excision and programmed nuclear death by autophagy, which has enriched and broadened knowledge of those mechanisms. PMID- 21624460 TI - Downregulation of pre-rRNA processing gene Mamrd1 decreases growth, conidiation and virulence in the entomopathogenic fungus Metarhizium acridum. AB - Mrd1 is one of the trans-acting proteins and plays an important role in precursor ribosomal RNA processing. Here we characterized the Mamrd1 gene from Metarhizium acridum and studied its function in growth, conidiation and virulence using RNA interference. The Mamrd1 gene was identified as participating in the processing of pre-rRNA in M. acridum and was highly upregulated during the infection process. A Mamrd1-RNAi strain exhibited an appearance of fluffy mycelia, a defective branching pattern and delayed conidiation compared to the wild-type strain. Downregulation of Mamrd1 in M. acridum suppressed growth both on artificial medium and inside the insect, and significantly reduced hyphal biomass, conidium production and virulence against Locusta migratoria manilensis. These results demonstrated that Mamrd1 plays an important role in growth, conidiation and virulence in M. acridum. PMID- 21624461 TI - An immune-induced reeler protein is involved in the Bombyx mori melanization cascade. AB - In this study, we isolated two reeler cDNAs from bacteria-challenged larval fat bodies of the silkworm, Bombyx mori. A reeler domain spanned most of the coding regions of these two cDNAs, and their expression patterns were different in B. mori larval tissues. The reeler1 gene was strongly induced by Escherichia coli K12 and Bacillus subtilis in B. mori larval hemocytes, fat bodies and midguts, but reeler2 was expressed at extremely low levels in these tissues. We focused on the reeler1 gene for functional analysis. Interference by double-stranded reeler1 RNA in vivo led to reduced nodule formation in bacteria-injected larvae, while the injection of recombinant Reeler1 promoted nodule formation in reeler1 gene silenced larvae, indicating that Reeler1 is involved in the nodulation response. Knockdown of the reeler1 gene significantly decreased phenoloxidase activity in bacteria-challenged larval hemolymph, while injection of recombinant Reeler1 enhanced phenoloxidase activity, suggesting that Reeler1 is involved in the prophenoloxidase activation cascade. Our results provide new mechanistic evidence about the melanization cascade in the insect immunity. PMID- 21624462 TI - A polymorphism of NADPH oxidase p22 phox is associated with reduced susceptibility to acute rejection in renal allograft recipients. AB - BACKGROUND: Reactive oxygen species (ROS) such as superoxide (O(2)(-)) play important roles in inflammatory processes. By altering the redox environment, ROS modulate the activation of transcription factors and cytokine genes involved in acute cellular rejection. The NAD(P)H oxidase is a multi-subunit enzyme present in leucocytes and endothelial cells, and is a key source of O(2)(-). 3 single nucleotide polymorphisms (SNPs) of the p22 phox subunit were investigated in a large cohort of renal allograft recipients. METHODS: The C242T, A640G and A-930G SNPs were studied in 244 Caucasian patients with end stage renal failure (ESRF) (148 renal allograft recipients and 96 dialysis patients) using standard PCR. Acute rejection was diagnosed by renal biopsy in 66 allograft recipients (44.6%). Normal controls were DNA samples extracted from 131 umbilical cord bloods following uncomplicated obstetric delivery. RESULTS: A highly significant increase in the frequency of the T242 allele in patients with ESRF compared to controls (31.3% vs 16.7%, p<0.0001) and in allograft recipients without acute rejection compared to those with rejection (37.8% vs 27.3%, p<0.0001) was demonstrated. CONCLUSION: These results show that the T242 allele may predispose to the development of ESRF, but paradoxically reduce susceptibility to acute rejection through reduced NAD(P)H oxidase activity. PMID- 21624463 TI - Serotonergic mediated body mass index changes in Parkinson's disease. AB - More than 50% of patients with Parkinson's disease (PD) are expected to show abnormalities with their weight in a process that starts several years before the diagnosis. The serotonergic (5-HT) system has been proposed to regulate appetite and the 5-HT transporter (SERT) is a key modulator of 5-HT metabolism. Here, we hypothesized that a dysfunctional 5-HT system could be responsible for alterations of weight in PD and we sought to investigate this in vivo. Thirty four PD patients had Body Mass Index (BMI) changes monitored over a 12-month period and one positron emission tomography (PET) brain scan with (11)C-DASB, a selective marker of SERT availability, during their second clinical assessment. Results were compared with those of a group of 10 normal controls. Half (17) of the PD patients showed abnormal BMI changes over the 12-month period; 12 lost while 5 gained weight. PD patients with abnormal BMI changes showed significantly raised (11)C-DASB binding in rostral raphe nuclei, hypothalamus, caudate nucleus and ventral striatum compared to cases with no significant BMI changes. (11)C DASB binding in other regions was similarly decreased in the PD BMI subgroups compared to normal controls. BMI gainers showed significantly raised (11)C-DASB binding in anterior cingulate cortex (ACC) compared to BMI losers. Our findings suggest that abnormal BMI changes over a 12-month period are linked with relatively raised SERT availability in PD on an overall background of decreased 5 HT function. The regions implicated are the rostral raphe nuclei and its connections to limbic and cognitive areas. It is conceivable that 5-HT agents could help alleviate abnormal changes in BMI in PD. PMID- 21624464 TI - Bcl2-A1 interacts with pro-caspase-3: implications for amyotrophic lateral sclerosis. AB - Expression of mutant SOD1 typical of familial amyotrophic lateral sclerosis (ALS) induces the expression of Bcl2-A1, a member of the Bcl2 family of proteins, specifically in motor neurons of transgenic mice. In this work, we have used immortalized motor neurons (NSC-34) and transgenic mice expressing mutant SOD1 to unravel the molecular mechanisms and the biological meaning of this up regulation. We report that up-regulation of Bcl2-A1 by mutant SOD1 is mediated by activation of the redox sensitive transcription factor AP1 and that Bcl2-A1 interacts with pro-caspase-3 via its C-terminal helix alpha9. Furthermore, Bcl2 A1 inhibits pro-caspase-3 activation in immortalized motor neurons expressing mutant SOD1 and thus induction of Bcl2-A1 in ALS mice represents a pro-survival strategy aimed at counteracting the toxic effects of mutant SOD1. These data provide significant new insights on how molecular signaling, driven by expression of the ALS-causative gene SOD1, affects regulation of apoptosis in motor neurons and thus may have implications for ALS therapy, where prevention of motor neuronal cell death is one of the major aims. PMID- 21624465 TI - Rescue of a dystrophin-like protein by exon skipping normalizes synaptic plasticity in the hippocampus of the mdx mouse. AB - Duchenne muscular dystrophy (DMD) is caused by the absence of dystrophin, a protein that fulfills important functions in both muscle and brain. The mdx mouse model of DMD, which also lacks dystrophin, shows a marked reduction in gamma aminobutyric acid type A (GABA(A))-receptor clustering in central inhibitory synapses and enhanced long-term potentiation (LTP) at CA3-CA1 synapses of the hippocampus. We have recently shown that U7 small nuclear RNAs modified to encode antisense sequences and expressed from recombinant adeno-associated viral (rAAV) vectors are able to induce skipping of the mutated exon 23 and to rescue expression of a functional dystrophin-like product both in the muscle and nervous tissue in vivo. In the brain, this rescue was accompanied by restoration of both the size and number of hippocampal GABA(A)-receptor clustering. Here, we report that 25.2+/-8% of re-expression two months after intrahippocampal injection of rAAV reverses the abnormally enhanced LTP phenotype at CA3-CA1 synapses of mdx mice. These results suggests that dystrophin expression indirectly influences synaptic plasticity through modulation of GABA(A)-receptor clustering and that re expression of the otherwise deficient protein in the adult can significantly alleviate alteration of neural functions in DMD. PMID- 21624466 TI - Interleukin 4 induces the apoptosis of mouse microglial cells by a caspase dependent mechanism. AB - Microglial cells are resident macrophages in the central nervous system (CNS) and become activated in many pathological conditions. Activation of microglial cells results in reactive microgliosis, manifested by an increase in cell number in the affected CNS regions. The control of microgliosis may be important to prevent pathological damage to the brain. The type 2 cytokine IL-4 has been reported to be protective in brain inflammation. However, its effect on microglial cell survival was not well understood. In this study, we report a dual effect of IL-4 on the survival of mouse microglial cells. In a 6h short term culture, IL-4 reduced the death of microglial cells induced by staurosporine. In contrast, in long term treatment (more than 48h), IL-4 increased the apoptotic death of both primary mouse microglial cells and a microglial cell line N9. Mechanistic studies revealed that, in microglial cells, IL-4 increased the levels of cleaved caspase 3 and PARP, which is down-stream of activated caspase 3. In addition, IL-4 down regulated the autophagy and the antiapoptotic protein Bcl-xL in microglial cells. On the other hand, the pre-incubation of microglial cells with IL-4 for 24h, attenuated the cell death induced by the neurotoxic peptide amyloid beta 1-42 (Abeta42). Our observations demonstrate a novel function of IL-4 in regulating the survival of microglial cells, which may have important significance in reduction of undesired inflammatory responses in the CNS. PMID- 21624467 TI - Systemic delivery of NEMO binding domain/IKKgamma inhibitory peptide to young mdx mice improves dystrophic skeletal muscle histopathology. AB - The activation of nuclear factor kappaB (NF-kappaB) contributes to muscle degeneration that results from dystrophin deficiency in human Duchenne muscular dystrophy (DMD) and in the mdx mouse. In dystrophic muscle, NF-kappaB participates in inflammation and failure of muscle regeneration. Peptides containing the NF-kappaB Essential Modulator (NEMO) binding domain (NBD) disrupt the IkappaB kinase complex, thus blocking NF-kappaB activation. The NBD peptide, which is linked to a protein transduction domain to achieve in vivo peptide delivery to muscle tissue, was systemically delivered to mdx mice for 4 or 7 weeks to study NF-kappaB activation, histological changes in hind limb and diaphragm muscle and ex vivo function of diaphragm muscle. Decreased NF-kappaB activation, decreased necrosis and increased regeneration were observed in hind limb and diaphragm muscle in mdx mice treated systemically with NBD peptide, as compared to control mdx mice. NBD peptide treatment resulted in improved generation of specific force and greater resistance to lengthening activations in diaphragm muscle ex vivo. Together these data support the potential of NBD peptides for the treatment of DMD by modulating dystrophic pathways in muscle that are downstream of dystrophin deficiency. PMID- 21624468 TI - Gastrointestinal dysfunction contributes to weight loss in Huntington's disease mice. AB - Weight loss is the most important non-neurological complication of Huntington's disease (HD). It correlates with disease progression and affects the quality of life of HD patients, suggesting that it could be a valuable target for therapeutic intervention. The mechanism underlying weight loss in HD is unknown. Mutant huntingtin, the protein that causes the disease, is not only expressed in the brain, but also along the gastrointestinal (GI) tract. Here we demonstrate that the GI tract of HD mice is affected. At the anatomical level we observed loss of enteric neuropeptides, as well as decreased mucosal thickness and villus length. Exploring the functions of the GI system we found impaired gut motility, diarrhea, and malabsorption of food. The degree of malabsorption was inversely associated with body weight, suggesting that GI dysfunction plays an important role in weight loss in HD mice. In summary, these observations suggest that the GI tract is affected in HD mice and that GI dysfunction contributes to nutritional deficiencies and weight loss. PMID- 21624469 TI - Temporal changes in mRNA expression of the brain nutrient transporters in the lithium-pilocarpine model of epilepsy in the immature and adult rat. AB - The lithium-pilocarpine model mimics most features of human temporal lobe epilepsy. Following our prior studies of cerebral metabolic changes, here we explored the expression of transporters for glucose (GLUT1 and GLUT3) and monocarboxylates (MCT1 and MCT2) during and after status epilepticus (SE) induced by lithium-pilocarpine in PN10, PN21, and adult rats. In situ hybridization was used to study the expression of transporter mRNAs during the acute phase (1, 4, 12 and 24h of SE), the latent phase, and the early and late chronic phases. During SE, GLUT1 expression was increased throughout the brain between 1 and 12h of SE, more strongly in adult rats; GLUT3 increased only transiently, at 1 and 4h of SE and mainly in PN10 rats; MCT1 was increased at all ages but 5-10-fold more in adult than in immature rats; MCT2 expression increased mainly in adult rats. At all ages, MCT1 and MCT2 up-regulation was limited to the circuit of seizures while GLUT1 and GLUT3 changes were more widespread. During the latent and chronic phases, the expression of nutrient transporters was normal in PN10 rats. In PN21 rats, GLUT1 was up-regulated in all brain regions. In contrast, in adult rats GLUT1 expression was down-regulated in the piriform cortex, hilus and CA1 as a result of extensive neuronal death. The changes in nutrient transporter expression reported here further support previous findings in other experimental models demonstrating rapid transcriptional responses to marked changes in cerebral energetic/glucose demand. PMID- 21624470 TI - Pharmacokinetic and chemoprevention studies on tea in humans. AB - Green tea and its major polyphenols constituents, tea catechins, have been shown to have many health benefits including cancer prevention. Tea catechins and tea catechin metabolites/catabolites are bioavailable in the systemic circulation after oral intake of green tea or green tea catechins. The metabolites/catabolites identified in humans include glucuronide/sulfate conjugates, methylated tea catechin conjugates, and microflora-mediated ring fission products and phenolic acid catabolites. Plasma levels of unchanged tea catechins in humans are mostly in the sub-MUM or nM concentration range, which is much lower than the effective concentrations determined in most in vitro studies. However, some of the catechin metabolites/catabolites are present in the systemic circulation at levels much higher than those of the parent catechins. The contribution of catechin derived metabolites/catabolites to the biological effects associated with green tea is yet to be defined. A limited number of chemoprevention trials of green tea or green tea catechins have been conducted to date and have observed potential preventive activity for oral, prostate, and colorectal cancer. Emerging data from multiple ongoing intervention trials will further contribute to defining the cancer preventive activity of green tea or green tea catechins. PMID- 21624471 TI - Matrix metalloproteinase inhibition with tetracyclines for the treatment of coronary artery disease. AB - Coronary artery disease is caused by atherosclerosis - a progressive arterial inflammatory disease that is responsible for significant global mortality and morbidity through the development of the acute coronary syndromes: sudden cardiac death, acute myocardial infarction and unstable angina. These clinical entities share a common pathophysiology: rupture of atherosclerotic plaque resulting in abrupt complete or partial thrombotic obstruction of coronary blood flow. Matrix metalloproteinases (MMP), through their central role in tissue remodeling and inflammation, are secreted by inflammatory cells of the atherosclerotic plaque and are capable of degrading all the extracellular matrix components of the fibrous cap that separates the atherosclerotic lesion from blood flow in the arterial lumen. Plaque rupture occurs when the circumferential tensile stresses in the artery overwhelm the structural integrity of the progressively degraded, thinned and weakened fibrous cap of the atherosclerotic lesion. Tetracyclines inhibit MMPs through their ability to chelate zinc. Subantimicrobial doses of doxycycline have been shown to reduce inflammation and inhibit MMP activity in patients with coronary artery disease. Further investigation is warranted to assess the potential clinical risks and benefits of MMP inhibition with tetracyclines or other agents in the treatment of coronary artery disease. PMID- 21624472 TI - Gut CaVP is an innate immune protein against bacterial challenge in amphioxus Branchiostoma belcheri. AB - The importance of calcium-binding proteins in immune response of vertebrates is determined, but whether they have the role in invertebrates is largely unknown. In the present study, phylogenetic analysis indicated that calcium vector protein (CaVP), a protein unique to amphioxus, shared 68% similarity in amino acid sequence with human and mouse calmodulin (CaM). CaVP cDNA was cloned into a bacterial vector pET-32a, and its His-tagged fusion protein was produced in Eschherichia coli cells (BL21). The recombinant CaVP was purified by Ni-NTA column and SDS-PAGE, and then utilized for antibody preparing. The prepared antibodies could recognize amphioxus CaVP with high specificity. Further analysis by Western blotting showed that CaVP was detected in muscle and humoral fluid of normal animals and appeared in gut of bacterial immunized or challenged amphioxus. Interestingly, gut CaVP was significantly higher in a healthy sub group than a wounded sub-group post bacterial challenge. This response was detected strongly in immunization and challenge by the same Gram-negative bacterium Vibro parahaemolyticus and weakly in immunization by V. parahaemolyticus and then challenge by Gram-negative Aeromonas hydrophila, whereas no any feedback was found in immunization by V. parahaemolyticus and challenge by Gram-positive Staphylococcus aureus. These findings indicate the importance of gut CaVP in response to bacterial challenge. PMID- 21624473 TI - In vivo imaging of mitochondrial function in methamphetamine-treated rats. AB - Abuse of the powerfully addictive psychostimulant, methamphetamine, occurs worldwide. Recent studies have suggested that methamphetamine-induced dopaminergic neurotoxicity is related to oxidative stress. In response to nerve activation, the mitochondrial respiratory chain is rapidly activated. The enhancement of mitochondrial respiratory chain activation may induce oxidative stress in the brain. However, there is little experimental evidence regarding the mitochondrial function after methamphetamine administration in vivo. Here, we evaluated whether a single administration of methamphetamine induces ATP consumption and overactivation of mitochondria. We measured mitochondrial function in two different ways: by monitoring oxygen partial pressure using an oxygen-selective electrode, and by imaging of redox reactions using a nitroxyl radical (i.e., nitroxide) coupled with Overhauser-enhanced magnetic resonance imaging (OMRI). A single administration of methamphetamine to Wistar rats induced dopaminergic nerve activation, ATP consumption and an increase in mitochondrial respiratory chain function in both the striatum and cortex. Furthermore, antioxidant TEMPOL prevented the increase in mitochondrial oxidative damage and methamphetamine-induced sensitization. These findings suggest that energy supplying reactions after dopaminergic nerve activation are associated with oxidative stress in both the striatum and cortex, leading to abnormal behavior. PMID- 21624474 TI - Effect of external auditory pacing on the neural activity of stuttering speakers. AB - External auditory pacing, such as metronome sound and speaking in unison with others, has a fluency-enhancing effect in stuttering speakers. The present study investigated the neural mechanism of the fluency-enhancing effect by using functional magnetic resonance imaging (fMRI). 12 stuttering speakers and 12 nonstuttering controls were scanned while performing metronome-timed speech, choral speech, and normal speech. Compared to nonstuttering controls, stuttering speakers showed a significantly greater increase in activation in the superior temporal gyrus under both metronome-timed and choral speech conditions relative to a normal speech condition. The caudate, globus pallidus, and putamen of the basal ganglia showed clearly different patterns of signal change from rest among the different conditions and between stuttering and nonstuttering speakers. The signal change of stuttering speakers was significantly lower than that of nonstuttering controls under the normal speech condition but was raised to the level of the controls, with no intergroup difference, in metronome-timed speech. In contrast, under the chorus condition the signal change of stuttering speakers remained lower than that of the controls. Correlation analysis further showed that the signal change of the basal ganglia and motor areas was negatively correlated with stuttering severity, but it was not significantly correlated with the stuttering rate during MRI scanning. These findings shed light on the specific neural processing of stuttering speakers when they time their speech to auditory stimuli, and provide additional evidence of the efficacy of external auditory pacing. PMID- 21624475 TI - Simultaneous monitoring of intracellular pH changes and hemodynamic response during cortical spreading depression by fluorescence-corrected multimodal optical imaging. AB - Cortical spreading depression (CSD) plays an important role in trauma, migraine and ischemia. CSD could induce pronounced hemodynamic changes and the disturbance of pH homeostasis which has been postulated to contribute to cell death following ischemia. In this study, we described a fluorescence-corrected multimodal optical imaging system to simultaneously monitor CSD associated intracellular pH (pH(i)) changes and hemodynamic response including hemoglobin concentrations and cerebral blood flow (CBF). CSD was elicited by application of KCl on rat cortex and direct current (DC) potential was recorded as a typical characteristic of CSD. The pH(i) shift was mapped by neutral red (NR) fluorescence which was excited at 516-556 nm and emitted at 625 nm. The changes in hemoglobin concentrations were determined by dual-wavelength optical intrinsic signal imaging (OISI) at 550 nm and 625 nm. Integration of fluorescence imaging and dual-wavelength OISI was achieved by a time-sharing camera equipped with a liquid crystal tunable filter (LCTF). CBF was visualized by laser speckle contrast imaging (LSCI) through a separate camera. Besides, based on the dual-wavelength optical intrinsic signals (OISs) obtained from our system, NR fluorescence was corrected according to our method of fluorescence correction. We found that a transient intracellular acidification followed by a small alkalization occurred during CSD. After CSD, there was a prolonged intracellular acidification and the recovery of pH(i) from CSD took much longer time than those of hemodynamic response. Our results suggested that the new multimodal optical imaging system had the potential to advance our knowledge of CSD and might work as a useful tool to exploit neurovascular coupling under physiological and pathological conditions. PMID- 21624476 TI - Ventral striatal and medial prefrontal BOLD activation is correlated with reward related electrocortical activity: a combined ERP and fMRI study. AB - Functional magnetic resonance imaging (fMRI) research suggests that the ventral striatum (VS)/nucleus accumbens, medial prefrontal cortex (mPFC), and broader mesocorticolimbic dopamine system mediate aspects of reward processing from expectation of reward to pleasantness experienced upon reward attainment. In parallel, research utilizing event-related potentials (ERP) indicates that the feedback negativity (FN) is sensitive to reward vs. non-reward feedback and outcome expectation. The FN has been source localized to the mPFC and dorsal striatum, and converging evidence suggests that the FN reflects reward processing in the mesocorticolimbic system. However, the extent to which ERP and fMRI measures of reward processing are correlated has yet to be explored within the same individuals. The primary aim of the current study was to examine the convergence between fMRI (i.e., VS and mPFC) and ERP (i.e., FN) measures of reward processing in forty-two participants who completed counterbalanced fMRI and ERP sessions while performing the same monetary gambling task. For the Win>Loss comparison, fMRI activation in the mesocorticolimbic reward circuit including the VS and mPFC was positively correlated with the FN. Here, we demonstrate that monetary gains activate the VS, mPFC, caudate, amygdala, and orbital frontal cortex, enhance the FN ERP component within 300 ms post feedback, and that these measures are related. Thus, fMRI and ERP measures provide complementary information about mesocorticolimbic activity during reward processing, which may be useful in assessing pathological reward sensitivity. PMID- 21624477 TI - The relationship between fibrogenic TGFbeta1 signaling in the joint and cartilage degradation in post-injury osteoarthritis. AB - OBJECTIVE: To review the literature on modulation of chondrocyte activities in the osteoarthritic joint, and to discuss these changes in relation to established hard and soft tissue repair paradigms, with an emphasis on transforming growth factor beta (TGFbeta1)-mediated signaling which can promote either a chondrogenic or fibrogenic phenotype. METHODS: Papers addressing the close relationship between repair in general, and the specific post-injury response of joint tissues are summarized. Different interpretations of the role of TGFbeta1 in the emergence of an "osteoarthritic" chondrocyte are compared and the phenotypic plasticity of "reparative" progenitor cells is examined. Lastly, emerging data on a central role for A-Disintegrin-And-Metalloproteinase-with-Thrombospondin-like Sequences-5 (ADAMTS5) activity in modulating TGFbeta1 signaling through activin receptor-like kinase 1 (ALK1) and activin receptor-like kinase 5 (ALK5) pathways is discussed. RESULTS: The review illustrates how a transition from ALK5-mediated fibrogenic signaling to ALK1-mediated chondrogenic signaling in joint cells represents the critical transition from a non-reparative to a reparative cell phenotype. Data from cell and in vivo studies illustrates the mechanism by which ablation of ADAMTS5 activity allows the transition to reparative chondrogenesis. Multiple large gene expression studies of normal and osteoarthritis (OA) human cartilages (CAs) also support an important role for TGFbeta1-mediated pro fibrogenic activities during disease progression. CONCLUSIONS: We conclude that progressive articular CA damage in post-injury OA results primarily from biomechanical, cell biologic and mediator changes that promote a fibroblastic phenotype in joint cells. Since ADAMTS5 and TGFbeta1 appear to control this process, agents which interfere with their activities may not only enhance endogenous CA repair in vivo, but also improve the properties of tissue engineered CA for implantation. PMID- 21624478 TI - Snorc is a novel cartilage specific small membrane proteoglycan expressed in differentiating and articular chondrocytes. AB - OBJECTIVE: Maintenance of chondrocyte phenotype is a major issue in prevention of degeneration and repair of articular cartilage. Although the critical pathways in chondrocyte maturation and homeostasis have been revealed, the in-depth understanding is deficient and novel modifying components and interaction partners are still likely to be discovered. Our focus in this study was to characterize a novel cartilage specific gene that was identified in mouse limb cartilage during embryonic development. METHODS: Open access bioinformatics tools and databases were used to characterize the gene, predicted protein and orthologs in vertebrate species. Immunohistochemistry and mRNA expression methodology were used to study tissue specific expression. Fracture callus and limb bud micromass culture were utilized to study the effects of BMP-2 during experimental chondrogenesis. Fusion protein with C-terminal HA-tag was expressed in Cos7 cells, and the cell lysate was studied for putative glycosaminoglycan attachment by digestion with chondroitinase ABC and Western blotting. RESULTS: The predicted molecule is a small, 121 amino acids long type I single-pass transmembrane chondroitin sulfate proteoglycan, that contains ER signal peptide, lumenal/extracellular domain with several threonines/serines prone to O-N acetylgalactosamine modification, and a cytoplasmic tail with a Yin-Yang site prone to phosphorylation or O-N-acetylglucosamine modification. It is highly conserved in mammals with orthologs in all vertebrate subgroups. Cartilage specific expression was highest in proliferating and prehypertrophic zones during development, and in adult articular cartilage, expression was restricted to the uncalcified zone, including chondrocyte clusters in human osteoarthritic cartilage. Studies with experimental chondrogenesis models demonstrated similar expression profiles with Sox9, Acan and Col2a1 and up-regulation by BMP-2. Based on its cartilage specific expression, the molecule was named Snorc, (Small NOvel Rich in Cartilage). CONCLUSION: A novel cartilage specific molecule was identified which marks the differentiating chondrocytes and adult articular chondrocytes with possible functions associated with development and maintenance of chondrocyte phenotype. PMID- 21624479 TI - Noradrenergic enhancement of associative fear memory in humans. AB - Ample evidence in animals and humans supports the noradrenergic modulation in the formation of emotional memory. However, in humans the effects of stress on emotional memory are traditionally investigated by declarative memory tests (e.g., recall, recognition) for non-associative emotional stimuli (e.g., stories, pictures). Given that anxiety disorders are thought to originate from associative learning processes and are characterized by distressing emotional responses, the existing literature seems to be inconclusive for the understanding of these disorders. Here, we tested whether noradrenaline strengthens the emotional expression of associative fear memory by using a differential fear conditioning procedure in humans. Stimulation of the noradrenergic system by the administration of yohimbine HCl (20mg) during memory formation did not directly augment the differential startle fear response 48 h later. Yet, the other retention tests uncovered that the administration of yohimbine HCl contrary to placebo pill extensively delayed the process of extinction learning and generated a superior recovery of fear (i.e., reinstatement and reacquisition). Conversely, the yohimbine HCl manipulation did not affect the skin conductance responding and the US expectancy ratings, emphasizing the concept of multiple memory systems. To our knowledge this is the first demonstration in humans that increased noradrenaline release during or shortly after a stressful event strengthens the formation of associative fear memory traces. The present findings suggest that noradrenaline may play an important role in the etiology and maintenance of anxiety disorders. PMID- 21624480 TI - Cognitive deficits in Rett syndrome: what we know and what we need to know to treat them. AB - Rett syndrome is an autism spectrum disorder and a leading cause of severe mental retardation in girls. The nature of the cognitive abnormalities in Rett, as described in humans and other animal models, and its potential reversibility and treatment are the subject of this review. Rett syndrome is associated with severe mental retardation and a host of impairments that include social and motor deficits, and respiratory and bone abnormalities. More than 80% of Rett girls have mutations in the gene that encodes MeCP2, which is a protein with a complex set of functions that include transcriptional repression and activation. The complex phenotype associated with Rett and the knowledge of the causal genetic mutation provide a unique opportunity within the autism spectrum to explore the relationship between transcriptional control, brain abnormalities and specific behavioral functions, importantly the elusive cognitive dysfunctions associated with mental retardation. The nature of the cognitive abnormalities related to Rett and the potential reversibility and treatment of these abnormalities have not been studied as extensively as some of the other aspects of the Rett phenotype. The cognitive phenotype associated with Rett is also less well studied relative to that in other well known developmental disorders, such as Down syndrome and Fragile X. Nevertheless, some recent studies provide hope that the cognitive impairments, as well as other symptoms of Rett, can be rescued. PMID- 21624481 TI - Contribution of the parafascicular nucleus in the spontaneous object recognition task. AB - The parafascicular (PF) nucleus, a posterior component of the intralaminar nuclei of the thalamus, is considered to be an essential structure in the feedback systems of basal ganglia-thalamo-cortical circuits critically involved in cognitive processes. The specific role played by multimodal information encoded in PF neurons in learning and memory processes is still unclear. We conducted two experiments to investigate the role of the PF in the spontaneous object recognition (SOR) task. The behavioral effects of pretraining rats with bilateral lesions of PF with N-methyl-D-aspartate (NMDA) were compared to vehicle controls. In the first experiment, rats were tested on their ability to remember the association immediately after training trials and in the second experiment after a 24h delay. Our findings provide evidence that PF lesions critically affect both SOR tests and support its role in that non-spatial form of relational memory. PMID- 21624482 TI - Towards mouse models of perseveration: a heritable component in extinction of operant behavior in fourteen standard and recombinant inbred mouse lines. AB - Extinction of instrumental responses is an essential skill for adaptive behavior such as foraging. So far, only few studies have focused on extinction following appetitive conditioning in mice. We studied extinction of appetitive operant lever-press behavior in six standard inbred mouse strains (A/J, C3H/HeJ, C57BL/6J, DBA/2J, BALB/cByJ and NOD/Ltj) and eight recombinant inbred mouse lines. From the response rates at the end of operant and extinction training we computed an extinction index, with higher values indicating better capability to omit behavioral responding in absence of reward. This index varied highly across the mouse lines tested, and the variability was partially due to a significant heritable component of 12.6%. To further characterize the relationship between operant learning and extinction, we calculated the slope of the time course of extinction across sessions. While many strains showed a considerable capacity to omit responding when lever pressing was no longer rewarded, we found a few lines showing an abnormally high perseveration in lever press behavior, showing no decay in response scores over extinction sessions. No correlation was found between operant and extinction response scores, suggesting that appetitive operant learning and extinction learning are dissociable, a finding in line with previous studies indicating that these forms of learning are dependent on different brain areas. These data shed light on the heritable basis of extinction learning and may help develop animal models of addictive habits and other perseverative disorders, such as compulsive food seeking and eating. PMID- 21624483 TI - Probiotic (symbiotic) bacterial languages. AB - Symbiotic gut microorganisms release of various soluble low molecular weight (LMW)molecules of different chemical nature (surface and exogenous proteins, nucleases, serpins, sirtuines, other enzymes, lectins, peptides, amines, bacteriocines, fatty and amino acids, lactones, furanons, miRNA, NO, etc). These LMW molecules are able to sense environment, interact with corresponding cell surface, membrane, cytoplasm and nucleic acid receptors, to reply quickly and coordinately by induction of special sets of genes, to support stability of host genome and microbiome, to modulate epigenomic regulation of gene phenotypic expression, to ensure the information exchange in numerous bacterial and bacteria host systems playing an important role in the control for many genetic and physiological functions, biochemical and behaviour reactions, in supporting host health in general. Various symbiotic (probiotic) strains produce different spectrum of such LMW molecules. There is chemical and functional similarity between LMW molecules synthesized by host eukaryotic cells, indigenous and probiotic microorganisms and some micronutrients. It means many LMW compounds of different origin may be the universal regulators contributing to the transmission of information, quorum sensing effects, metagenome stability and epigenomic control for cell growth and development as well as phenotypic expression of different genes. Knowledge accumulated concerning molecular languages of symbiotic microorganisms allows us to better understand the mode of action of known probiotics and to design in principle novel probiotics (metabiotics) with increased health effectiveness. Now we are only at the beginning of a new era of molecular personal biotherapy and nutrition. Soon we can successfully manipulate both the host and its microbiota through interfering in their cross talk, stability and epigenomic regulation of expression of genes using various types of eukaryotic, prokaryotic and nutrition origin LMW molecules are capable to modulate genetic, metabolic and physiological activities. PMID- 21624484 TI - In vitro assay of the antimicrobial activity of kephir against bacterial and fungal strains. AB - Kephir is a fermented carbonated refreshing milk, with a slightly acidic aromatic taste and creamy foam composition which contains lactobacilli, leuconostocci, acetic acid bacteria, lactostreptococci and yeasts. Recent studies have demonstrated its antibacterial, immunostimulating, antitumoral and cholesterol lowering activities. PURPOSE: The purpose of this study was to investigate the antimicrobial activity of kephir against Bacillus subtilis spp. spizizenii ATCC 6633, Staphylococcus aureus ATCC 6538, Enterococcus faecalis ATCC 29212, Escherichia coli ATCC 8739, Salmonella enteritidis ATCC 13076, Pseudomonas aeruginosa ATCC 9027 and Candida albicans ATCC 10231. The kephir fermented for 24 h and 48 h, as well and after 7 days preservation at 4-8 degrees C was tested by in vitro disk diffusion method. The intensity of the antimicrobial activity was interpreted by comparison with two antibiotics, i.e. ampicillin and neomycin. RESULTS: The antimicrobial activity of 24 h as well as 48 fermented kephir, fresh or after 7 days preservation at 4-8 degrees C was similar and observed against B. subtilis, S. aureus, E. coli, E. faecalis and S. enteritidis. For E. coli, E. faecalis and S. enteritidis the antimicrobial activity was superior to both tested antibiotics and for B. subtilis and S. aureus to one antibiotic. The tested products exhibited no activity against P. aeruginosa and C. albicans. CONCLUSION: Kephir is exhibiting large spectrum and strong antibacterial activity probably due to the complex viable probiotic strains association producing antimicrobial substances. PMID- 21624485 TI - Surface-enhanced laser desorption ionization/time-of-flight (SELDI-TOF) mass spectrometry (MS) as a phenotypic method for rapid identification of antibiotic resistance. AB - Based on experiments with 10 defined strains of Escherichia coli, we present a new method for bacterial phenotyping using SELDI-TOF mass spectrometry. Changes in bacterial protein profiles in the context of the time of cultivation and the antibiotic environment were minimal. Proteom subprofiling may further distinguish between strains with specific susceptibility to antimicrobials. Mass spec-based methods may become common in the future of bacterial pathogen identification in clinical microbiology diagnostics. PMID- 21624486 TI - Effect of obesity on outcomes after autologous hematopoietic stem cell transplantation for multiple myeloma. AB - Obesity has implications for chemotherapy dosing and selection of patients for therapy. Autologous hematopoietic stem cell transplant (AutoHCT) improves outcomes for patients with multiple myeloma, but optimal chemotherapy dosing for obese patients is poorly defined. We analyzed the outcomes of 1087 recipients of AutoHCT for myeloma reported to the CIBMTR between 1995 and 2003 who received high-dose melphalan conditioning, with or without total body irradiation (TBI). We categorized patients by body mass index (BMI) as normal, overweight, obese, or severely obese. There was no overall effect of BMI on progression-free survival (PFS), overall survival (OS), progression, or nonrelapse mortality (NRM). In patients receiving melphalan and TBI conditioning, obese and severely obese patients had superior PFS and OS compared with normal and overweight patients, but the clinical significance of this finding is unclear. More obese patients were more likely to receive a reduced dose of melphalan, but there was no evidence that melphalan or TBI dosing variability affected PFS. Therefore, current common strategies of dosing melphalan do not impair outcomes for obese patients, and obesity should not exclude patients from consideration of autologous transplantation. Further research is necessary to optimize dosing of both chemotherapy and radiation in obese patients. PMID- 21624487 TI - Tubulogenesis during blood vessel formation. AB - The ability to form and maintain a functional system of contiguous hollow tubes is a critical feature of vascular endothelial cells (ECs). Lumen formation, or tubulogenesis, occurs in blood vessels during both vasculogenesis and angiogenesis in the embryo. Formation of vascular lumens takes place prior to the establishment of blood flow and to vascular remodeling which results in a characteristic hierarchical vessel organization. While epithelial lumen formation has received intense attention in past decades, more recent work has only just begun to elucidate the mechanisms controlling the initiation and morphogenesis of endothelial lumens. Studies using in vitro and in vivo models, including zebrafish and mammals, are beginning to paint an emerging picture of how blood vessels establish their characteristic morphology and become patent. In this article, we review and discuss the molecular and cellular mechanisms driving the formation of vascular tubes, primarily in vivo, and we compare and contrast proposed models for blood vessel lumen formation. PMID- 21624488 TI - Cell culture models using rat primary alveolar type I cells. AB - There is a lack of cell culture models using primary alveolar type I (AT I) cells. The purpose of this study was to develop cell culture models using rat AT I cells and microvascular endothelial cells from the lung (MVECL). Two types of model systems were developed: single and co-culture systems; additionally a 3 dimensional model system was developed. Pure AT I cell (96.3 +/- 2.7%) and MVECL (97.9 +/- 1.1%) preparations were used. AT I cell morphology, mitochondrial number and distribution, actin filament arrangement and number of apoptotic cells at confluence, and telomere attrition were characterized. AT I cells maintained their morphometric characteristics through at least population doubling (PD) 35, while demonstrating telomere attrition through at least PD 100. Furthermore, AT I cells maintained the expression of their specific markers, T1alpha and AQ-5, through PD 42. For the co-cultures, AT I cells were grown on the top and MVECL were grown on the bottom of fibronectin-coated 24-well Transwell FluroblokTM filter inserts. Neither cell type transmigrated the 1 MUm pores. Additionally, AT I cells were grown in a thick layer of Matrigel((r)) to create a 3-dimensional model in which primary AT I cells form ring-like structures that resemble an alveolus. The development of these model systems offers the opportunities to investigate AT I cells and their interactions with MVECL in response to pharmacological interventions and in the processes of disease, repair and regeneration. PMID- 21624489 TI - L-NAME and L-arginine differentially ameliorate cigarette smoke-induced emphysema in mice. AB - Nitric oxide (NO) represents one of the most important intra- and extracellular mediators and takes part in both biologic and pathologic processes. This study aimed to verify the treatment with an NO inhibitor and an NO substrate in pulmonary emphysema induced by cigarette smoke (CS) in a murine model. We compared N-acetylcysteine (NAC), a precursor of glutathione, to G-nitro-L arginine-methyl ester or L-NAME (LN), which is an NO inhibitor, and to l-arginine (LA), which is a substrate for NO formation. Mice were divided into several groups: control, CS, CS + LN, CS + LA, and CS + NAC. Control and CS groups were treated daily with a vehicle, while CS + LN, CS + LA, and CS + NAC groups were treated daily with LN (60 mg/kg), LA (120 mg/kg) and NAC (200 mg/kg), respectively. The bronchoalveolar lavage was analyzed and the lungs were removed for histological and biochemical analysis. CS increases neutrophil number. Neutrophil number was lowest in CS + LN, followed by CS + LA. The lungs of CS + LN, CS + LA and CS + NAC mice were protected compared to the lungs of CS mice, but not equal to the quality of lungs in control mice. The CS group also exhibited increased oxidative stress, which was also present in the CS + LN group and to a lesser extent in the CS + LA group. Tissue inhibitor of metalloproteinase 1 and 2 increased in the CS + LN group and to a lesser extent in the CS + LA group relative to the control group. These results suggest that LN and LA treatment protected the mouse lung from CS. However, NAC treatment was more than LN and LA. We suggest that the protection conferred by LN treatment requires a balance between proteases and antiproteases, and that protection conferred by LA treatment involves the balance between oxidants and antioxidants. PMID- 21624490 TI - Comparison of budesonide/formoterol Turbuhaler with fluticasone/salmeterol Diskus for treatment effects on small airway impairment and airway inflammation in patients with asthma. AB - BACKGROUND: A course of combination therapy with an inhaled corticosteroid (ICS) and a long-acting beta(2) agonist (LABA) for asthma can improve lung function, asthma symptoms and reduce exacerbations. Because both medicinal substance and inhalation devices are associated with clinical efficacy, each ICS/LABA combination may have different features. This study aimed to compare the effects of two widely available formulations, budesonide/formoterol (BUD/FM) delivered by a Turbuhaler((r)), and fluticasone/salmeterol (FP/SM) delivered by a Diskus((r)), on small airway function and airway inflammation. METHODS: Asthmatic patients (n = 40) treated twice daily with FP/SM 250/50 MUg with forced expiratory volume in 1 s values controlled above 80% of the predicted normal but with suspected persistent airway inflammation and small airway impairment were enrolled in the study. Patients were randomized into two groups, receiving either twice daily BUD/FM 320/9 MUg or FP/SM 250/50 MUg, and treatment efficacy was compared after 4 weeks. Outcomes included impulse oscillometry (IOS), fractional exhaled nitric oxide (FeNO), spirometry and Asthma Control Questionnaire (ACQ) scores. RESULTS: Patients in the BUD/FM group showed significant improvements in their IOS and spirometry parameters of small airway function, FeNO values and ACQ scores, compared with the FP/SM group. There were good correlations between IOS parameters, FeNO and ACQ score changes over the course of the treatment. CONCLUSIONS: BUD/FM twice daily significantly improved small airway impairment and airway inflammation in asthmatic patients, leading to a reduction in asthma symptoms and achievement of good asthma control. In addition, improvement of small airway function may improve airway inflammation and/or lead to better controlled asthma. PMID- 21624491 TI - Safety and tolerability of an oral MMP-9 and -12 inhibitor, AZD1236, in patients with moderate-to-severe COPD: a randomised controlled 6-week trial. AB - BACKGROUND: Matrix metalloproteinases (MMPs) have been implicated in the pathogenesis of chronic obstructive pulmonary disease (COPD). This phase IIa study investigated the safety and tolerability of oral AZD1236, an MMP-9 and MMP 12 inhibitor, in patients with COPD. Efficacy analyses were included on an exploratory basis. METHODS: This was a multinational, randomised, double-blind, parallel-group study conducted in 74 men and women aged >=40 years with stable moderate-to-severe COPD. After a 2-week run-in period, patients received oral AZD1236 75 mg, or matching placebo, twice daily for 6 weeks (on top of background medication with short-acting bronchodilators and/or inhaled corticosteroids, if applicable). In addition to safety and tolerability endpoints (AEs, vital signs and laboratory assessments) efficacy was assessed as a secondary objective (spirometry, 6MWT, BODE index and biomarkers in blood and urine. Clinical COPD Questionnaire (CCQ), peak expiratory flow (PEF) and daily diary cards of symptom severity were completed by the patients. RESULTS: The incidence of adverse events (AEs) was similar in AZD1236 and placebo recipients; 28 in 13 patients (37%) and 21 in 17 patients (44%), respectively; the difference in actual number reported accounted for primarily by mild AEs in the AZD1236 group. The most commonly experienced AEs in both groups were COPD exacerbations, headache and viral infections. One patient in the AZD1236 group experienced a serious AE of interstitial nephritis (comprising of acute renal failure, rash, fever and blood eosinophilia) considered to be related to treatment. After 6 weeks, AZD1236 had demonstrated no significant effect on lung function, 6MWT, BODE index or biomarkers compared with placebo. No meaningful differences were observed in patient-reported CCQ score, PEF, COPD symptoms or use of rescue medication. CONCLUSIONS: For most of these COPD patients, with the particular exception of one who experienced a serious AE, AZD1236 at 75 mg twice daily was generally well tolerated and had an acceptable safety profile. Therapeutic efficacy could not be demonstrated, possibly due to the stable disease and background medications of the patients enrolled in this small, short-term study. PMID- 21624492 TI - Association of the variants in AGT gene with modified drug response in Korean aspirin-intolerant asthma patients. AB - The angiotensinogen (AGT) gene enhances the effect of several bronchoconstrictors and produces a peptide that is accumulated in the airways of asthma patients; events that may underpin the pathogenesis of aspirin-intolerant asthma (AIA). To carry out a case-control analysis between AGT and aspirin-induced bronchospasm following treatment with an anti-asthma drug, montelukast (MLK), 38 single nucleotide polymorphisms (SNPs) in AGT were genotyped in 56 AIA cohort. Genotyping was performed with TaqMan assay and haplotypes were inferred using PHASE algorithm ver. 2.0. Statistical analyses of each SNPs and haplotypes were performed using SAS version 9.1. Among 13 variants displaying significant signals, two SNPs (+2401C>G and +2476C>T) in the intronic region of AGT were significantly associated with modification of drug response even after correction for multiple testing (P=0.0009-0.002; P(corr)=0.02-0.03). Furthermore, the two variants also exhibited associations with MLK response rate (P=0.0003-0.0006; P(corr)=0.006-0.01). Although our results are preliminary and further replication in a larger-scale group of subjects should be warranted, these observations provide evidence that AGT variants might be one of genetic factors involved in the response of anti-asthma drugs in AIA patients. PMID- 21624493 TI - The influence of dietary supplementation of arachidonic acid on prostaglandin production and oxidative stress in the Pacific oyster Crassostrea gigas. AB - In a previous study, dietary supplementation with arachidonic acid (ARA) to oysters Crassostrea gigas increased haemocyte numbers, phagocytosis, and production of reactive oxygen species level (ROS) by haemocytes (Delaporte et al., 2006). To assess if the observed stimulation of these cellular responses resulted from changes of ARA-related prostaglandin (PG) production, we analysed prostaglandin E2 metabolite (PGEM) content on the same oysters fed three levels of ARA. Dietary supply of polyunsaturated fatty acids (PUFA) could also induce an oxidative stress that could similarly increase cellular responses; therefore, two indicators of oxidative stress were analysed: peroxidation level and antioxidant defence status. Together the observed positive correlation between ARA and PGEM levels and the absence of lipid peroxidation and antioxidant activity changes supports the hypothesis of an immune stimulation via PG synthesis. Although ARA proportion in oyster tissues increased by up to 7-fold in response to ARA dietary supplementation, peroxidation index did not change because of a compensatory decrease in n-3 fatty acid proportion, mainly 22:6n-3. To further confirm the involvement of PG in the changes of haemocyte count, phagocytosis and ROS production upon ARA supplementation, it would be interesting to test cyclooxygenase and lipooxygenase inhibitors in similar experiments. PMID- 21624494 TI - Multiple pathways from three types of sugar receptor sites to metabotropic transduction pathways of the blowfly: study by the whole cell-clamp experiments. AB - Multiple pathways from three types of multiple receptor sites to three types of metabotropic signal transduction pathways were investigated in the whole cell clamp experiments using isolated labellar sugar receptor neurons (cells) of the adult blowfly, Phormia regina. First, the concentration-response curves of three types of sweet taste components specialized to multiple receptor sites were obtained: sucrose for the pyranose sites (P-sites), fructose for the furanose sites (F-sites), and l-valine for the alkyl sites (R-sites). Next, the effects of inhibitors such as 2', 5'-dideoxyadenosine on adenylyl cyclase in the cAMP pathway, LY 83583 on guanylyl cyclase in the cGMP pathway, and U-73122 on phospholipase C in the IP3 pathway were examined. The results showed that all of the inhibitors affected each specific target in the second-messenger transduction pathways. The obtained results verified that the P-site corresponded to the cAMP, the F-site to the cGMP, and the R-site to the IP3 transduction pathway, and that these three signal pathways did not have crossing points. PMID- 21624495 TI - Arginine kinase in Phytomonas, a trypanosomatid parasite of plants. AB - Phytomonas are trypanosomatid plant parasites closely related to parasites that cause several human diseases. Little is known about the biology of these organisms including aspects of their metabolism. Arginine kinase (E.C. 2.7.3.3) is a phosphotransferase which catalyzes the interconversion between the phosphagen phosphoarginine and ATP. This enzyme is present in some invertebrates and is a homolog of another widely distributed phosphosphagen kinase, creatine kinase. In this work, a single canonical arginine kinase isoform was detected in Phytomonas Jma by enzymatic activity assays, PCR, and Western Blot. This arginine kinase is very similar to the canonical isoforms found in T. cruzi and T. brucei, presenting about 70% of amino acid sequence identity and a very similar molecular weight (40kDa). The Phytomonas phosphagen system seems to be very similar to T. cruzi, which has only one isoform, or T. brucei (three isoforms); establishing a difference with other trypanosomatids, such as Leishmania, which completely lacks phosphagen kinases, probably by the presence of the arginine-consuming enzyme, arginase. Finally, phylogenetic analysis suggests that Kinetoplastids' arginine kinase was acquired, during evolution, from the arthropod vectors by horizontal gene transfer. PMID- 21624496 TI - Expression analysis and response of Penaeus monodon 14-3-3 genes to salinity stress. AB - Two isoforms of 14-3-3 protein, namely 14-3-3A and 14-3-3B, from the shrimp Penaeus monodon were investigated for their potential role in adaptation to salinity stress. Transcripts of 14-3-3A were found in various shrimp tissues whilst expression of 14-3-3B transcripts was more specific being observed in the osmoregulatory tissues, that is in the gills and epipodites. In shrimp gills, the 14-3-3A transcript levels slightly changed after transfer of the shrimp from 3 to 25 or to 40 ppt. In contrast, significant change in the mRNA levels in response to salinity stress was detected for 14-3-3B, where a significant decrease of 14-3 3B transcript was observed in gills of shrimp transferred from hypo-osmotic (3 ppt) salinity to iso-osmotic or hyper-osmotic (25 and 40ppt, respectively) salinity. On the other hand, shrimp transferred from 40 ppt to 3 ppt showed a strong induction of 14-3-3B mRNA expression in the gills. These transcript expression analyses suggest that 14-3-3B is likely to be involved in the hyper osmotic regulation in P. monodon. In addition, 14-3-3B appeared to regulate ATPase function since suppression of the gene by RNA interference resulted in a significant decrease in the total ATPase activity. PMID- 21624497 TI - Enhanced glycoprotein production in HEK-293 cells expressing pyruvate carboxylase. AB - There is an imperative need for expression systems allowing the efficient and robust manufacturing of high quality glycoproteins. In the present work, HEK-293 cells stably expressing interferon-alpha2b were further engineered with the insertion of the yeast pyruvate carboxylase 2 gene. In batch cultures, marked reductions in lactate and ammonia production were observed compared to the parental cell clone. Although the maximum specific growth rate remained unchanged, the altered metabolism led to a 2-fold increase in maximum cell density and 33% increase in the integral of viable cell concentration and interferon production yield. The underlying metabolic changes were further investigated using various (13)C-labeled substrates and measuring the resulting lactate mass isotopomer distributions. Simultaneous metabolite and isotopomer balancing allowed the accurate determination of key intracellular fluxes. Such detailed and quantitative knowledge about the central carbon metabolism of the cells is instrumental to further support the development of high-yield fed-batch processes. PMID- 21624498 TI - Pharmaceutical drug transport: the issues and the implications that it is essentially carrier-mediated only. AB - All cells necessarily contain tens, if not hundreds, of carriers for nutrients and intermediary metabolites, and the human genome codes for more than 1000 carriers of various kinds. Here, we illustrate using a typical literature example the widespread but erroneous nature of the assumption that the 'background' or 'passive' permeability to drugs occurs in the absence of carriers. Comparison of the rate of drug transport in natural versus artificial membranes shows discrepancies in absolute magnitudes of 100-fold or more, with the carrier containing cells showing the greater permeability. Expression profiling data show exactly which carriers are expressed in which tissues. The recognition that drugs necessarily require carriers for uptake into cells provides many opportunities for improving the effectiveness of the drug discovery process. PMID- 21624499 TI - Electronic health records: Implications for drug discovery. AB - Electronic health records (EHRs) have increased in popularity in many countries. Pushed by legal mandates, EHR systems have seen substantial progress recently, including increasing adoption of standards, improved medical vocabularies and enhancements in technical infrastructure for data sharing across healthcare providers. Although the progress is directly beneficial to patient care in a hospital or clinical setting, it can also aid drug discovery. In this article, we review three specific applications of EHRs in a drug discovery context: finding novel relationships between diseases, re-evaluating drug usage and discovering phenotype-genotype associations. We believe that in the near future EHR systems and related databases will impact significantly how we discover and develop safe and efficacious medicines. PMID- 21624500 TI - FDA-approved drug labeling for the study of drug-induced liver injury. AB - Drug-induced liver injury (DILI) is a leading cause of drugs failing during clinical trials and being withdrawn from the market. Comparative analysis of drugs based on their DILI potential is an effective approach to discover key DILI mechanisms and risk factors. However, assessing the DILI potential of a drug is a challenge with no existing consensus methods. We proposed a systematic classification scheme using FDA-approved drug labeling to assess the DILI potential of drugs, which yielded a benchmark dataset with 287 drugs representing a wide range of therapeutic categories and daily dosage amounts. The method is transparent and reproducible with a potential to serve as a common practice to study the DILI of marketed drugs for supporting drug discovery and biomarker development. PMID- 21624501 TI - mTOR as a multifunctional therapeutic target in HIV infection. AB - Patients undergoing long-term highly active antiretroviral therapy treatment are probably at a higher risk of various HIV-related complications. Hyperactivation of The mammalian target of rapamycin (mTOR) has been found to contribute to dysregulated apoptosis and autophagy which determine CD4(+)-T-cell loss, impaired function of innate immunity and development of neurocognitive disorders. Dysregulated mTOR activation has also been shown to play a key part in the development of nephropathy and in the pathogenesis of HIV-associated malignancies. These studies strongly support a multifunctional key role for mTOR in the pathogenesis of HIV-related disorders and suggest that specific mTOR inhibitors could represent a novel approach for the prevention and treatment of these pathologies. PMID- 21624502 TI - A capsule or a tablet. PMID- 21624503 TI - Genetic diversity of Plasmodium vivax malaria in China and Myanmar. AB - Genetic diversity and population structure of Plasmodium vivax parasites are valuable to the prediction of the origin and spread of novel variants within and between populations, and to the program evaluation of malaria control measures. Using two polymorphic genetic markers, the merozoite surface protein genes PvMSP 3alpha and PvMSP-3beta, we investigated the genetic diversity of four Southeast Asian P. vivax populations, representing both subtropical and temperate strains with dramatically divergent relapse patterns. PCR amplification of PvMSP-3alpha and PvMSP-3beta genes detected three and four major size polymorphisms among the 235 infections examined, respectively, while restriction analysis detected 15 and 19 alleles, respectively. Samples from different geographical areas differed dramatically in their PvMSP-3alpha and PvMSP-3beta allele composition and frequency. Samples tended to cluster on the basis of their PCR-RFLP polymorphism. These results indicated that different parasite genotypes were circulating in each endemic area, and that geographic isolation may exist. Multiple infections were detected in all four parasite populations, ranging from 20.5% to 31.8%, strongly indicating that P. vivax populations were highly diverse and multiple clonal infections are common in these malaria-hypoendemic regions of Southeast Asia. PMID- 21624504 TI - The active contribution of Toll-like receptors to allergic airway inflammation. AB - Epithelia lining the respiratory tract represent a major portal of entry for microorganisms and allergens and are equipped with innate and adaptive immune signaling receptors for host protection. These include Toll-like receptors (TLRs) that recognize microbial components and evoke diverse responses in cells of the respiratory system. TLR stimulation by microorganism-derived molecules activates antigen presenting cells, control T helper (Th) 1, Th2, and Th17 immune cell differentiation, cytokine production by mast cells, and activation of eosinophils. It is clear that TLR are involved in the pathophysiology of allergic airway diseases such as asthma. Dendritic cells (DCs), a kind of antigen presenting cells, which play a key role in the induction of allergic airway inflammation, are privileged targets for pathogen associated molecular patterns (PAMPs). During the allergic responses, engagement of TLRs on DCs determines the Th2 polarization of the T cells. TLR signaling in mast cells increases the release of IL-5, and TLR activation of airway epithelial cells forces the generation of proallergic Th2 type of cytokines. Although these responses aim to protect the host, they may also result in inflammatory tissue damage in the airway. Under certain conditions, stimulation of TLRs, in particular, TLR9, may reduce Th2-dependent allergic inflammation by induction of Th1 responses. Therefore, understanding the complex regulatory roles of TLRs in the pathogenesis of allergic airway inflammation should facilitate the development of preventive and therapeutic measures for asthmatic patients. PMID- 21624506 TI - Neurophysiological and epigenetic effects of physical exercise on the aging process. AB - Aging is a gradual process during which molecular and cellular processes deteriorate progressively, often leading to such pathological conditions as vascular and metabolic disorders and cognitive decline. Although the mechanisms of aging are not yet fully understood, inflammation, oxidative damage, mitochondrial dysfunction, functional alterations in specific neuronal circuits and a restricted degree of apoptosis are involved. Physical exercise improves the efficiency of the capillary system and increases the oxygen supply to the brain, thus enhancing metabolic activity and oxygen intake in neurons, and increases neurotrophin levels and resistance to stress. Regular exercise and an active lifestyle during adulthood have been associated with reduced risk and protective effects for mild cognitive impairment and Alzheimer's disease. Similarly, studies in animal models show that physical activity has positive physiological and cognitive effects that correlate with changes in transcriptional profiles. According to numerous studies, epigenetic events that include changes in DNA methylation patterns, histone modification and alterations in microRNA profiles seem to be a signature of aging. Hence, insight into the epigenetic mechanisms involved in the aging process and their modulation through lifestyle interventions such as physical exercise might open new avenues for the development of preventive and therapeutic strategies to treat aging-related diseases. PMID- 21624505 TI - Toll like receptors in diseases of the lung. AB - The lung is in continuous contact with a diverse array of infectious agents, foreign antigens, and host-derived danger signals. To sample this expansive internal and external milieu, both resident myeloid and stromal/structure cells of the lung express a full complement of toll like receptors (TLRs) which recognize pathogen-associated molecular patterns (PAMPs) and endogenous danger associated molecular patterns (DAMPs). TLRs play a vital role in immune host defense against bacterial, mycobacterial, fungal, and viral pathogens of the lung. Additionally, TLRs contribute to disease pathogenesis in non-infectious pulmonary disorders, including airway disease, acute lung injury, and interstitial lung disease. In this review, TLR biology in the context of experimental infectious and non-infectious lung disease is discussed, and correlates to human lung disease, including therapeutic implications of these findings, are defined. PMID- 21624507 TI - A clear look at the neuroimmunology of multiple sclerosis and beyond. AB - The term neuroimmunology was first coined to refer to a generic involvement of the immune system in the pathogenesis of neurological diseases, particularly of the central nervous system. Since then, the neuroimmunology spectrum has steadily grown and currently spans from classical autoimmune diseases of the central and peripheral nervous systems to previously unsuspected conditions such as autism spectrum disorders or chronic fatigue syndrome. Multiple sclerosis remains the predominant entity in terms of research efforts and social pressure as well as a good model of organ-specific autoimmune disease with limited therapeutic options. While the fast-pace genome-wide association studies reported a number of genes to be significantly associated with multiple sclerosis, these currently explain only a minor part of disease susceptibility. Further, clinicians are continuously challenged with the clinical classifications of immune-mediated or autoimmune central and peripheral conditions and with other pragmatic questions such as the roles of vaccination and physical therapy. For these reasons the present collection of Autoimmunity Reviews is timely as it will address these major issues related to neuroimmunology. PMID- 21624508 TI - Expression and characterization of an extremely thermostable beta-glycosidase (mannosidase) from the hyperthermophilic archaeon Pyrococcus furiosus DSM3638. AB - Genomic analysis of the hyperthermophilic archaeon Pyrococcus furiosus revealed the presence of an open reading frame (ORF PF0356) similar to the enzymes in glycoside hydrolase family 1. This beta-glycosidase, designated PFTG (P. furiosus thermostable glycosidase), was cloned and expressed in Escherichia coli. The expressed enzyme was purified by heat treatment and Ni-NTA affinity chromatography. The gene was composed of 1,452 bp encoding 483 amino acids for a protein with a predicted molecular mass of 56,326 Da. The temperature and pH optima were 100 degrees C and 5.0 in sodium citrate buffer, respectively. The substrate specificity of PFTG suggests that it possesses characteristics of both beta-galactosidase and beta-mannosidase activities. However, through kinetic studies by ITC (Isothermal Titration Colorimetry) which is very sensitive method for enzyme kinetics, PF0356 enzyme revealed the highest catalytic efficiency toward p-nitrophenyl-beta-d-mannopyranoside (3.02 k(cat)/K(m)) and mannobiose (4.32 k(cat)/K(m)). The enzyme showed transglycosylation and transgalactosylation activities toward cellobiose, lactose and mannooligosaccharides that could produce GOS (galactooligosaccharides) and MOS (maltooligosaccharides). This novel hyperthermostable beta-glycosidase may be useful for food and pharmaceutical applications. PMID- 21624509 TI - Hepatoprotection of quercetin against oxidative stress by induction of metallothionein expression through activating MAPK and PI3K pathways and enhancing Nrf2 DNA-binding activity. AB - Flavonoids are natural phenolic substances widely found in fruit, vegetables, grains, and wine. Most of these compounds exert health-promoting effects seem to attribute to their antioxidant activity. Metallothioneins (MT) has been suggested to protect against acute heavy metal toxicity in the liver, and the proteins of MT can be induced by various stimuli including antioxidant. Measuring the induction of MT genes may provide an efficient approach to understand the chemopreventive mechanisms of flavonoids. The antioxidant activity of eight flavonoids was determined by TEAC and ORAC assays and their effects on MT protein were also measured. HepG2 cells were employed to explore the mechanisms underlying flavonoid-induced MT induction. Statistical analysis revealed a positive correlation between the antioxidant activity of flavonoids and MT expression. Quercetin-induced MT expression may function by activating the phosphorylation of JNK, p38 and PI3K/Akt as well as by enhancing Nrf2 DNA-binding activity. Moreover, quercetin exhibited a potential protective effect on t-BHP caused injury in hepatocytes through the induction of MT. These results suggest that quercetin is a natural antioxidant in the diet and the consumption of foods that are rich in quercetin could be beneficial for the prevention of environmental oxidant-induced liver damage. PMID- 21624510 TI - Verification of a biomarker discovery approach for detection of Down syndrome in amniotic fluid via multiplex selected reaction monitoring (SRM) assay. AB - Prenatal screening test for Down syndrome (DS) can be improved by discovery of novel biomarkers. A multiplex selected reaction monitoring (SRM) assay was developed to test previously identified thirteen candidate proteins in amniotic fluid (AF). One unique peptide was selected for each protein based on discovery data, while three MS/MS transitions were selected based on intelligent SRM results. For one of the candidates, matrix metalloproteinase-2 (MMP2), ELISA was also performed to validate SRM results in AF and to test serum samples. Comparison of AF samples from DS versus controls via SRM assay revealed five proteins that were differentially expressed. Bile salt-activated lipase, mucin 13, carboxypeptidase A1, and dipeptidyl peptidase 4 showed a decrease in DS affected AF, and MMP2 showed an increase, in comparison to controls (P<0.05). Discovery-based spectral counting ratios and SRM ratios showed a strong correlation, and MMP2 ELISA further confirmed the validity of the SRM data. Potential implications of differentially expressed proteins during fetal development are proposed. Our data also shows that SRM can provide a high throughput and accurate platform for biomarker verification. PMID- 21624511 TI - Influencing safe perioperative practice through communication. PMID- 21624512 TI - Accountable care: is it solely cost-conscious? PMID- 21624513 TI - Clarification on mask use during droplet precautions. PMID- 21624514 TI - Apply or sponsor someone for an AORN Award. PMID- 21624516 TI - Understanding our link in the chain of instrument care. PMID- 21624518 TI - Clarification on information technology roles. PMID- 21624525 TI - Perioperative pharmacology: blood coagulation modifiers. AB - Blood coagulation is the process that results in the formation of a blood clot to stop bleeding from a damaged blood vessel. Various pharmacologic agents can affect the coagulation process. The American College of Chest Physicians' evidence-based practice guidelines for perioperative management of antithrombotic therapy provide guidance for anticoagulant or antiplatelet therapy and bridge therapy. Perioperative nurses must understand the pharmacologic principles of the most common blood coagulation modifiers related to perioperative use. The perioperative nurse's responsibilities regarding administration of blood coagulation modifiers include reviewing the patient's pertinent laboratory results (eg, prothrombin time, partial thromboplastin time, international normalized ratio), recognizing the underlying conditions that require blood coagulation therapy, and documenting all pertinent information. Perioperative nurses also should participate in development of detailed storage and retrieval policies related to heparin. PMID- 21624526 TI - Transformational leadership: implications for nursing leaders in facilities seeking magnet designation. AB - A perioperative nurse leader's ability to effect positive change and inspire others to higher levels of achievement is related to his or her leadership style in the practice setting and the leadership style that is present across the organization. The American Nurses Credentialing Center's MagnetTM designation and redesignation process requires the demonstration of transformational leadership as one of the components of excellence. Transformational leadership can increase nurses' job satisfaction and commitment to the organization and organizational culture. Engaging staff members in the transition to transformational leadership and developing a common mission, vision, and goals are keys to success in the surgical setting. Bass's four interrelated leadership components-idealized influence, inspirational motivation, intellectual stimulation, and individual consideration-and associated behaviors were used by surgical services leaders in an East Coast, two-hospital system to successfully achieve redesignation as a Magnet facility. PMID- 21624527 TI - Warming blanket head drapes and trapped anesthetic gases: understanding the fire risk. AB - Polyethylene warming blanket head drapes are widely used to help surgical patients maintain normothermia. The OR quality management team at Froedtert Hospital, Milwaukee, Wisconsin, designed a quality improvement project using an intubation mannequin to determine whether a head drape used on an intubated patient would enhance the risk of ignition in the presence of an undetected anesthesia system gas leak. The team revealed several significant factors, including higher oxygen flow rate, the presence and application of the head drape, and not using the warming blanket blower, in the accumulation of oxidizers. Considerations for the surgical team when using the head drape include cutting a fenestration in the drape around the endotracheal tube or otherwise venting the drape, using the blanket blower, and having the anesthesia care provider frequently lift the head drape, thus minimizing the accumulation of trapped gases. PMID- 21624528 TI - AORN and University of Michigan School of nursing research alliance. AB - Research related to perioperative care requires advanced training and is well suited to take place at a research-intensive university. A recent research alliance established between AORN and the University of Michigan School of Nursing, Ann Arbor, uses the strengths of both a robust perioperative professional organization and a research-intensive university to make progress toward improving patient safety and transforming the perioperative work environment. Research activities undertaken by this alliance include investigating nurse staffing characteristics and patient outcomes, as well as evaluating the congruence and definitions of data elements contained in AORN's SYNTEGRITYTM Standardized Perioperative Framework. Disseminating the findings of the alliance is expected to facilitate the communication and application of new knowledge to nursing practice and help advance the perioperative nursing profession. PMID- 21624529 TI - AORN Ergonomic Tool 4: Solutions for Prolonged Standing in Perioperative Settings. AB - Prolonged standing during surgical procedures poses a high risk of causing musculoskeletal disorders, including back, leg, and foot pain, which can be chronic or acute in nature. Ergonomic Tool 4: Solutions for Prolonged Standing in Perioperative Settings provides recommendations for relieving the strain of prolonged standing, including the use of antifatigue mats, supportive footwear, and sit/stand stools, that are based on well-accepted ergonomic safety concepts, current research, and access to new and emerging technology. PMID- 21624530 TI - Tablet and e-reader technology in health care and education. PMID- 21624531 TI - Perioperative care of the patient with diastolic heart failure. PMID- 21624532 TI - Path to green: practice improvement in the OR. PMID- 21624533 TI - The normalization of deviance: what are the perioperative risks? PMID- 21624534 TI - Stepping up state certification priorities. PMID- 21624536 TI - Tacit handover, overt mishap. PMID- 21624537 TI - Prognostic usefulness of free fatty acids in patients with stable coronary heart disease. AB - Circulating nonesterified or free fatty acids (FFAs) may contribute to the development of cardiovascular pathology and correlate with ischemia in acute cardiovascular conditions. The aim of this study was to assess whether serum levels of FFAs are associated with long-term prognosis in subjects with stable coronary heart disease. This observational prospective cohort study included 1,206 participants in 3-weeks inpatient rehabilitation programs after acute myocardial infarction, coronary syndromes, or coronary intervention at 2 rehabilitation clinics in Germany (1999 to 2000). Eight-year prognosis (time to a secondary fatal or nonfatal cardiovascular disease event including myocardial infarction and stroke [n = 153] and time to death from any cause [n = 124]) was examined according to FFA quartiles and in spline regression. FFAs were correlated with established serum markers of cardiovascular risk and strongly related to secondary cardiovascular events and all-cause mortality in age- and gender-adjusted analysis. When additionally controlling for multiple established risk factors and risk markers, the hazard ratio in the fourth versus first quartile was 1.34 (95% confidence interval 0.79 to 2.24) for secondary cardiovascular events and 1.09 (95% confidence interval 0.62 to 1.91) for all cause mortality. Dose-response modeling suggested that very high FFAs might predict an increased risk for mortality (hazard ratio 1.98, 95% confidence interval 0.98 to 4.02, for 95th percentile vs first quartile). In conclusion, FFAs are closely correlated with cardiovascular risk markers, and in particular, very high FFA might identify patients with stable coronary heart disease with worse prognoses. PMID- 21624538 TI - Thirty-year (1975 to 2005) trends in the incidence rates, clinical features, treatment practices, and short-term outcomes of patients <55 years of age hospitalized with an initial acute myocardial infarction. AB - Sparse data are available describing recent trends in the magnitude, clinical features, treatment practices, and outcomes of comparatively young adults hospitalized with acute myocardial infarction (AMI). The objectives of this population-based study were to describe 3 decade-long trends (1975 to 2005) in these end points in adults <55 years old who were hospitalized with an initial AMI. The study population consisted of 1,703 residents of the Worcester (Massachusetts) metropolitan area 25 to 54 years of age who were hospitalized with initial AMIs at all central Massachusetts medical centers during 15 annual periods from 1975 through 2005. Overall hospital incidence rate (per 100,000 residents) of initial AMI in our study population was 66 (95% confidence interval 63 to 69) and incidence rates of AMI decreased inconsistently over time. Patients hospitalized during the most recent study years were more likely to have important cardiovascular risk factors and co-morbidities present but were less likely to have developed heart failure during their index hospitalization. In hospital and 30-day death rates decreased by approximately 50% (p = 0.04) during the years under study concomitant with increasing use of effective cardiac therapies. In conclusion, the results of this community-wide investigation provide insights into the magnitude, changing characteristics, and short-term outcomes of comparatively young patients hospitalized with a first AMI. Decreasing odds of developing or dying from an initial AMI during the 30 years under study likely reflect increased primary and secondary prevention and treatment efforts. PMID- 21624539 TI - Relation of maternal anti-Ro/La antibodies to aortic dilation in patients with congenital complete heart block. AB - An association between congenital complete atrioventricular block (cCAVB) and aortic dilation during childhood has recently been reported. We sought to further explore this relation with particular emphasis on the natural history of aortic abnormalities over time. The relation of maternal anti-Ro/La antibody status to the aortic size of children affected with cCAVB was also assessed. The patients were evaluated longitudinally with serial echocardiography. During a 15-year period, 62 patients at our institution were diagnosed with cCAVB, of whom 40% were exposed to maternal autoimmune antibodies and 35% were not. The antibody status in the remaining patients was unknown. The patients underwent 9.3 +/- 6.5 echocardiograms during the follow-up period. Dilation of the ascending aorta, defined as a z score >2.0, was present on the initial echocardiogram in all patients exposed to maternal antibodies and persisted during long-term follow-up in 96% of these patients. In contrast, 5% and 10% of patients without exposure to maternal autoimmune antibodies had aortic dilation on the initial and follow-up studies, respectively (p <0.001 and p <0.001, respectively). In conclusion, patients with autoimmune-mediated cCAVB merit periodic echocardiographic monitoring into adulthood to assess persistent or progressive aortic dilation and its attendant complications. PMID- 21624540 TI - Comparison of prevalence, clinical course, and pathological findings of left ventricular systolic impairment versus normal systolic function in patients with hypertrophic cardiomyopathy. AB - Impaired left ventricular systolic function (ILVSF) in hypertrophic cardiomyopathy (HC) is a risk factor for sudden death and a determinant of high mortality. We determined its prevalence, clinical parameters, long-term outcome, and pathologic findings of explanted hearts. We retrospectively analyzed 382 patients with HC; ILVSF was characterized by LV ejection fraction <50% at rest and was identified in 24 patients (6.3%). Patients with ILVSF were younger than patients with normal SF (43.5 +/- 14.1 vs 55.3 +/- 20.4 years, p = 0.001) and had larger LV end-diastolic cavity diameter (53.2 +/- 12.2 vs 43.8 +/- 6.2 mm, p = 0.001), larger left atrium (51.2 +/- 6.5 vs 44.3 +/- 8 mm, p <0.001), and lower fractional shortening (30.7 +/- 11.1% vs 45.5% +/- 10.3%, p <0.001). A combined end point (heart failure death or heart transplantation) was considered. Median follow-up was 3 years (1.2 to 6.3). Fourteen patients with ILVSF (58.3%) had the end point compared to 3 (0.8%) with normal SF (p <0.001). In explanted hearts, fibrosis represented 30.5 +/- 12.5% of the left ventricle; we observed a direct correlation between fibrosis and ventricular dilation (r = 0.794, p = 0.001) and an inverse correlation between fibrosis and ejection fraction (r = -0.623, p = 0.023). Number and length density of small arterioles (<50 MUm in diameter) were significantly decreased. In conclusion, ILVSF in HC has a poor prognosis and is associated with fibrosis and selective decreased development of small arterioles. PMID- 21624541 TI - Effects of chronic omega-3 polyunsaturated fatty acid supplementation on human pulmonary vein and left atrial electrophysiology in paroxysmal atrial fibrillation. AB - Omega-3 polyunsaturated fatty acids in fish oils may have antifibrillatory effects; however, their electrophysiologic effects in paroxysmal atrial fibrillation (PAF) remain unknown. The aim of this study was to investigate the effects of chronic fish oil supplementation on human pulmonary vein (PV) and left atrial electrophysiology in PAF. Patients with PAF undergoing PV isolation were randomized >=1 month before their procedure into a control group (n = 18) or a fish oil group (n = 18) in an unblinded fashion. The latter were supplemented with fish oil 6 g/day for a mean of 40 +/- 12 days. Pulmonary venous and left atrial effective refractory periods (ERPs), PV conduction, and susceptibility to AF initiated within PVs were assessed. Compared to the control group, the fish oil group had (1) longer left-sided (p = 0.002) and right-sided (p = 0.001) pulmonary venous ERPs; (2) less dispersion of pulmonary venous ERPs (left PVs p = 0.001, right PVs p = 0.07); (3) longer left atrial ERPs (p = 0.02); (4) no difference in pulmonary venous conduction; (5) lower incidence of AF initiated from PVs during ERP testing (77% vs 31%, p = 0.02); and (6) prolongation of mean AF cycle length (p = 0.009) and shortest AF cycle length in PVs (p = 0.04). In conclusion, patients with PAF chronically supplemented with fish oils exhibit distinctive electrophysiologic properties including prolonged pulmonary venous and left atrial ERPs and decreased susceptibility to initiation AF from within PVs. These changes may in part explain the antifibrillatory effect of chronic omega-3 polyunsaturated fatty acid supplementation in patients with PAF. PMID- 21624542 TI - Differences in cardiovascular risk profile between electrocardiographic hypertrophy versus strain in asymptomatic patients with aortic stenosis (from SEAS data). AB - Electrocardiograms are routinely obtained in clinical follow-up of patients with asymptomatic aortic stenosis (AS). The association with aortic valve, left ventricular (LV) response to long-term pressure load, and clinical covariates is unclear and the clinical value is thus uncertain. Data from clinical examination, electrocardiogram, and echocardiogram in 1,563 patients in the Simvastatin and Ezetimibe in Aortic Stenosis (SEAS) study were used. Electrocardiograms were Minnesota coded for arrhythmias and atrioventricular and intraventricular blocks; LV hypertrophy was assessed by Sokolow-Lyon voltage and Cornell voltage-duration criteria; and strain by T-wave inversion and ST-segment depression. Degree of AS severity was evaluated by echocardiography as peak aortic jet velocity and LV mass was indexed by body surface area. After adjustment for age, gender, LV mass index, heart rate, systolic and diastolic blood pressures, blood glucose, digoxin, antiarrhythmic drugs, drugs acting on the renin-angiotensin system, diuretics, beta blockers and calcium receptor blockers; peak aortic jet velocity was significantly greater in patients with electrocardiographic strain (mean difference 0.13 m/s, p <0.001) and LV hypertrophy by Sokolow-Lyon voltage criteria (mean difference 0.12 m/s, p = 0.004). After similar adjustment, LV mass index was significantly greater in patients with electrocardiographic strain (mean difference 14.8 g/cm(2), p <0.001) and LV hypertrophy by Sokolow-Lyon voltage criteria and Cornell voltage-duration criteria (mean differences 8.8 and 17.8 g/cm(2), respectively, p <0.001 for the 2 comparisons). In multiple comparisons patients with electrocardiographic strain had increased peak aortic jet velocity, blood glucose, and uric acid, whereas patients with LV hypertrophy by Sokolow-Lyon voltage criteria were younger and patients with LV hypertrophy by Cornell voltage-duration criteria more often were women. In conclusion, electrocardiographic criteria for LV hypertrophy and strain are independently associated with peak aortic jet velocity and LV mass index. Moreover, clinical covariates differ significantly between patients with electrocardiographic strain and those with LV hypertrophy by Sokolow-Lyon voltage criteria and Cornell voltage-duration criteria. PMID- 21624543 TI - Changes in emerging cardiac biomarkers after an intensive lifestyle intervention. AB - The present study evaluated the changes in emerging cardiac biomarkers, cognitive function, and social support measures after a comprehensive lifestyle intervention that included a low-fat, whole-foods, plant-based diet, exercise, stress management, and group support meetings. We conducted a prospective cohort study of 131 participants (59.2% women and 43.1% with diabetes mellitus), 56 with coronary heart disease (CHD) (37.5% women and 27.3% diabetes mellitus), and 75 at high risk with >=3 CHD risk factors and/or diabetes mellitus (76% women and 54.7% diabetes mellitus). The measurements were taken at baseline and 3 months after the intervention. Improvement in all targeted health behaviors was seen in both high-risk and CHD groups (all p <0.001) at 3 months, with reductions in body mass index, systolic and diastolic blood pressure, waist/hip ratio, C-reactive protein, insulin, low-density lipoprotein, high-density and total cholesterol, apolipoproteins A1 and B (all p <0.009) were observed. Nuclear magnetic resonance spectroscopy analysis of lipoprotein subclass particle concentrations and diameters showed a reduction in large very-low-density lipoprotein particles, size of the very-low-density lipoprotein particles, total low-density lipoprotein particles; total, large, and small high-density lipoprotein particles (all p <0.009) and small very-low-density lipoprotein particles (p <0.02). Increases in fibrinogen (p <0.03) and B-type natriuretic peptide (p <0.001) were seen, and these changes correlated inversely with the changes in the body mass index. The observed increase in B-type natriuretic peptide can be explained by the metabolic changes related to adipose tissue lipolysis. The quality of life, cognitive functioning, and social support measures significantly improved. In conclusion, lifestyle changes can be followed by favorable changes in traditional and emerging coronary heart disease biomarkers, quality of life, social support, and cognitive function among those with, or at high risk, of CHD. PMID- 21624544 TI - Low-dose electrocardiography synchronized nonenhanced computed tomography for assessing left atrium and pulmonary veins before radiofrequency catheter ablation for atrial fibrillation. AB - Recently contrast-enhanced cardiac computed tomography (CT) was found to be useful for imaging the left atrium and pulmonary veins (PVs) before radiofrequency catheter ablation in patients with atrial fibrillation. However, the risks of contrast agent in patients with impaired renal function must be considered. We investigated the accuracy of low-dose electrocardiographically synchronized nonenhanced cardiac CT (NECT) for identifying PV anatomy. One hundred eight consecutive patients who underwent cardiac CT before radiofrequency catheter ablation of atrial fibrillation were included. Nonenhanced cardiac computed tomogram was retrospectively evaluated for each patient by 2 radiologists for the following PV anatomy: conventional pattern, conjoined ostium, and accessory PVs with number and location. Sensitivity and specificity for variations in PVs were calculated using contrast-enhanced cardiac computed tomogram as the reference standard. Detection rates for each variation were also calculated. Twenty-one right PV (RPV) variations and 11 left PV (LPV) variations were observed. NECT showed a high diagnostic performance in detecting variations in PVs for the 2 observers. For RPV variations overall sensitivity was 97.6% and specificity was 96.6%. For LPV variations overall sensitivity was 90.9% and specificity was 97.9%. Overall detection rates for variation between the 2 observers were 97.1% for accessory RPV from the right middle lobe, 100% for 4 ostia with accessory RPV from the right middle lobe and accessory RPV from the superior segment of the right lower lobe, 100% for accessory RPV from the superior segment of the right lower lobe, 88.9% for conjoined ostium of the LPV, and 100% for accessory LPV from the left lingular segment. In conclusion, variations in PV anatomy were detected with great accuracy by NECT. PMID- 21624545 TI - Safety and results of cryoablation in patients <5 years old and/or <15 kilograms. AB - Current recommendations discourage elective radiofrequency ablation in patients <5 years old and/or weighing <15 kg, primarily because of the greater complication rate. To describe the current use, complications, and immediate outcomes of cryoablation in this patient population, a multicenter retrospective review of all patients <5 years old and/or weighing <15 kg who were treated with cryoablation for arrhythmia was performed. Eleven centers contributed data for 68 procedures on 61 patients. Of those, 34% were elective and 24% (n = 16) were both cryoablation and radiofrequency ablation. The median age and weight at ablation was 3.5 years (range 8 days to 9.9 years) and 15.2 kg (range 2.3 to 23), respectively. Congenital heart disease was present in 23% of the patients. The immediate success rate of cryoablation alone was 74%. No major complications occurred with cryoablation only; however, 2 of the 16 patients who underwent cryoablation and radiofrequency ablation had major complications. Of the 50 patients receiving cryoablation, 8 (16%) had variable degrees of transient atrioventricular block. The recurrence rate was 20% after cryoablation and 30% after cryoablation plus radiofrequency ablation. In conclusion, cryoablation appears to have a high safety profile in these patients. Compared to older and larger patients, the efficacy of cryoablation in this small, young population was lower and the recurrence rates were higher. Cryoablation's effect on the coronary arteries has not been fully elucidated and requires additional research. PMID- 21624546 TI - Impact of coronary calcium on outcome following sirolimus-eluting stent implantation. AB - There remain a small but sizable number of patients who develop restenosis after sirolimus-eluting stent (SES) implantation. However, the cause of SES restenosis has not been fully elucidated. The study population consisted of 52 patients with 69 lesions who underwent noninvasive coronary imaging by 64-slice multidetector computed tomography before SES deployment. Agatston calcium scores in target lesions were measured. All patients underwent follow-up coronary angiography at 8 months. Three coronary segments (in stent, proximal edge, and distal edge) were analyzed by quantitative coronary angiography. Agatston calcium score in target lesions averaged 214.7. Late lumen losses in the proximal edge, stent, and distal edge were 0.16 +/- 0.45, 0.47 +/- 0.58, and 0.07 +/- 0.29 mm, respectively. Lesions with restenosis at follow-up showed a trend to produce higher preprocedural calcium scores (629) compared to those without restenosis (153, p = 0.08). There was a significant positive correlation between lesion calcium score and in-stent late lumen loss (r = 0.47, p <0.01). In conclusion, assessment of coronary calcium by multidetector computed tomography might be useful to predict outcomes after SES implantation. PMID- 21624547 TI - Prenatal diagnosis of hypoplastic left heart syndrome in current era. AB - We sought to evaluate the relation of a prenatal diagnosis (preDx) with morbidity and mortality during the initial hospitalization in a contemporary cohort of patients with hypoplastic left heart syndrome (HLHS). A retrospective study of patients with HLHS presenting from 1999 to 2010 was performed. Patients with genetic disorders or a gestational age <34 weeks or who had intentionally received comfort care only were excluded. Of the 81 patients meeting the study criteria, 49 had a preDx and 32 were diagnosed postnatally (postDx). Birth weight (median 3.0 vs 3.4 kg; p = 0.007) and gestational age (median 38 vs 39 weeks; p <0.001) were lower in the preDx than in the postDx patients. Preoperatively, the postDx patients were intubated more frequently (97% vs 71%, p = 0.004) and ventilated longer (median 96 vs 24 hours, p = 0.005) than the preDx patients. They also had more preoperative acidosis, multiorgan failure, tricuspid valve regurgitation, and right ventricular dysfunction. Of the 73 patients undergoing surgery, no difference in survival was seen between the preDx and postDx groups (91% vs 89%). The median duration of postoperative ventilation was 7 days and the median length of stay was 36 days for the 66 survivors, with no difference between the 2 groups. Postoperative morbidities, including chylothorax and infection, were also similar in the preDx and postDx patients. No studied preoperative factor was associated with death, duration of postoperative ventilation, or length of stay. In conclusion, our recent experience has shown that preDx of HLHS was not associated with a survival advantage, fewer postoperative complications, or shorter length of stay. Improved preoperative status was observed in the preDx patients; however, they were born earlier with a lower birthweight. What effect these factors might have on longer term morbidity remains unknown. PMID- 21624548 TI - Usefulness of pericardial effusion as new diagnostic criterion for noninvasive detection of myocarditis. AB - Cardiovascular magnetic resonance (CMR) imaging holds promise for diagnosing myocarditis in vivo. The CMR diagnosis of myocarditis is determined by the ventricular morphology/function, late gadolinium enhancement, and T(2)-weighted imaging for myocardial edema. However, in routine clinical practice, we encounter patients with suspected myocarditis in the absence of left ventricular dysfunction, myocardial edema, or late gadolinium enhancement. In the present study, we sought to determine whether the presence of pericardial effusion could serve as a new diagnostic criterion and improve the sensitivity of CMR imaging to detect myocarditis. A total of 35 consecutive patients with biopsy proven virus associated myocarditis, onset of clinical symptoms within the past 3 months, and normal left ventricular function were enrolled in the present study. All patients underwent echocardiography, CMR imaging, and endomyocardial biopsy for workup of myocarditis. Late gadolinium enhancement was present in 16 patients (46%). Myocardial edema on T(2)-weighted imaging was present in 4 patients, but in just 1, it was the only abnormal finding. Pericardial effusion was present in 14 patients (40%). In 7 patients with myocarditis (20%), pericardial effusion was the only abnormal finding. Pericardial effusion, used as an additional diagnostic criterion, improved the sensitivity of CMR imaging for myocarditis from 46% to 66% (p = 0.023). In conclusion, pericardial effusion detected by CMR imaging might serve as a new diagnostic criterion for the noninvasive diagnosis of myocarditis in patients with recent onset of clinical symptoms and normal left ventricular function. PMID- 21624549 TI - Importance of diffuse atherosclerosis in the functional evaluation of coronary stenosis in the proximal-mid segment of a coronary artery by myocardial fractional flow reserve measurements. AB - The objective of this study was to evaluate the impact of diffuse coronary atherosclerosis on the functional evaluation of moderate coronary lesions in the proximal-mid segment of a coronary artery and its clinical implications. This was a prospective study including 100 consecutive patients with a moderate lesion (45 +/- 9% diameter stenosis) in the proximal-mid coronary segment who were evaluated with fractional flow reserve (FFR) measurement. No patient had any other angiographic stenosis distal to the evaluated coronary stenosis. FFR measurements were obtained just distal (~2 to 3 cm) to the lesion (FFR proximal measurement [FFR-PM]) and as distally as possible in the artery (FFR distal measurement [FFR DM]) after administration of the same dose of intracoronary adenosine. Thirty nine patients underwent dipyridamole or exercise myocardial single-photon emission computed tomography within 3 months of the FFR study. Mean FFR-PM was significantly higher compared to FFR-DM (0.84 +/- 0.08 vs 0.78 +/- 0.09, median gradient 0.06, 25th to 75th interquartile range 0.02 to 0.10, p <0.0001). FFR-DM was <0.75 in 33% of patients with FFR-PM >=0.75, leading to the decision of revascularization in these patients. Performing FFR measurement in the left main/left anterior descending artery predicted a higher gradient between FFR-DM and FFR-PM (odds ratio 4.58, 95% confidence interval 1.4 to 15.03, p = 0.007). FFR-DM exhibited a better correlation with results of myocardial single-photon emission computed tomography compared to FFR-PM (kappa 0.33 vs 0.22, p <0.0001). In conclusion, significant differences between FFR-DM and FFR-PM were observed in patients with moderate coronary stenosis in the proximal-mid segment of a coronary artery, with FFR-DM exhibiting a better correlation with results of noninvasive functional tests. These differences influenced the treatment decision in about 1/3 of patients and highlight the potential clinical relevance of coronary pressure wire positioning for functional evaluation of lesions in the proximal-mid segment of the coronary arteries. PMID- 21624550 TI - The link between erectile and cardiovascular health: the canary in the coal mine. AB - Lifestyle and nutrition have been increasingly recognized as central factors influencing vascular nitric oxide (NO) production and erectile function. This review underscores the importance of NO as the principal mediator influencing cardiovascular health and erectile function. Erectile dysfunction (ED) is associated with smoking, excessive alcohol intake, physical inactivity, abdominal obesity, diabetes, hypertension, and decreased antioxidant defenses, all of which reduce NO production. Better lifestyle choices; physical exercise; improved nutrition and weight control; adequate intake of or supplementation with omega-3 fatty acids, antioxidants, calcium, and folic acid; and replacement of any testosterone deficiency will all improve vascular and erectile function and the response to phosphodiesterase-5 inhibitors, which also increase vascular NO production. More frequent penile-specific exercise improves local endothelial NO production. Excessive intake of vitamin E, calcium, l-arginine, or l-citrulline may impart significant cardiovascular risks. Interventions discussed also lower blood pressure or prevent hypertension. Certain angiotensin II receptor blockers improve erectile function and reduce oxidative stress. In men aged <60 years and in men with diabetes or hypertension, erectile dysfunction can be a critical warning sign for existing or impending cardiovascular disease and risk for death. The antiarrhythmic effect of omega-3 fatty acids may be particularly crucial for these men at greatest risk for sudden death. In conclusion, by better understanding the complex factors influencing erectile and overall vascular health, physicians can help their patients prevent vascular disease and improve erectile function, which provides more immediate motivation for men to improve their lifestyle habits and cardiovascular health. PMID- 21624551 TI - Postnatal left ventricular diastolic function after fetal aortic valvuloplasty. AB - Fetal aortic balloon valvuloplasty (FAV) has shown promise in altering in utero progression of aortic stenosis to hypoplastic left heart syndrome. In patients who achieve a biventricular circulation after FAV, left ventricular (LV) compliance may be impaired. Echocardiographic indexes of diastolic function were compared between patients with biventricular circulation after FAV, congenital aortic stenosis (AS), and age-matched controls. In the neonatal period, patients with FAV had similar LV, aortic, and mitral valve dimensions but more evidence of endocardial fibroelastosis than patients with AS. Patients with FAV underwent more postnatal cardiac interventions than patients with AS (p = 0.007). Mitral annular early diastolic tissue velocity (E') was lower in patients with FAV and those with AS and controls in the neonatal period and over follow-up (p <0.001). Septal E' was similar among all 3 groups in the neonatal period. In follow-up patients, with FAV had lower septal E' than patients with AS or controls (p <0.001). Early mitral inflow velocity/E' was higher in patients with FAV as neonates and at follow-up (p <0.001). Mitral inflow pulse-wave Doppler-derived indexes of diastolic function were similar between groups. In conclusion, echocardiographic evidence of LV diastolic dysfunction is common in patients with biventricular circulation after FAV and persists in short-term follow-up. LV diastolic dysfunction in this unique population may have important implications on long-term risk of left atrial and subsequent pulmonary hypertension. PMID- 21624552 TI - Coronary artery distensibility assessed by 3.0 Tesla coronary magnetic resonance imaging in subjects with and without coronary artery disease. AB - Coronary vessel distensibility is reduced with atherosclerosis and normal aging, but direct measurements have historically required invasive measurements at cardiac catheterization. Therefore, we sought to assess coronary artery distensibility noninvasively using 3.0 Telsa coronary magnetic resonance imaging (MRI) and to test the hypothesis that this noninvasive technique can detect differences in coronary distensibility between healthy subjects and those with coronary artery disease (CAD). A total of 38 healthy, adult subjects (23 men, mean age 31 +/- 10 years) and 21 patients with CAD, diagnosed using x-ray angiography (11 men, mean age 57 +/- 6 years) were studied using a commercial whole-body MRI system. In each subject, the proximal segment of a coronary artery was imaged for the cross-sectional area measurements using cine spiral MRI. The distensibility (mm Hg(-1) * 10(3)) was determined as (end-systolic lumen area - end-diastolic lumen area)/(pulse pressure * end-diastolic lumen area). The pulse pressure was calculated as the difference between the systolic and diastolic brachial blood pressure. A total of 34 healthy subjects and 19 patients had adequate image quality for coronary area measurements. Coronary artery distensibility was significantly greater in the healthy subjects than in those with CAD (mean +/- SD 2.4 +/- 1.7 mm Hg(-1) * 10(3) vs 1.1 +/- 1.1 mm Hg(-1) * 10(3), respectively, p = 0.007; median 2.2 vs 0.9 mm Hg(-1) * 10(3)). In a subgroup of 10 patients with CAD, we found a significant correlation between the coronary artery distensibility measurements assessed using MRI and x-ray coronary angiography (R = 0.65, p = 0.003). In a group of 10 healthy subjects, the repeated distensibility measurements demonstrated a significant correlation (R = 0.80, p = 0.006). In conclusion, 3.0-Tesla MRI, a reproducible noninvasive method to assess human coronary artery vessel wall distensibility, is able to detect significant differences in distensibility between healthy subjects and those with CAD. PMID- 21624553 TI - Everolimus-eluting versus paclitaxel-eluting stents for treatment of bare metal stent restenosis. AB - First-generation drug-eluting stents have been proved to be very effective for the treatment of bare metal stent in-stent restenosis (BMS ISR). The efficacy of second-generation drug-eluting stents in this setting remains less well defined. The present study compared the long-term clinical outcome after treatment of BMS ISR using the second-generation everolimus-eluting stent (EES) to that after treatment using the paclitaxel-eluting stent (PES). A total of 174 patients with BMS ISR underwent percutaneous coronary intervention using a PES (95 patients) or an EES (79 patients) from 2003 to 2010. The patients in the PES and EES groups were followed up for 42.2 +/- 22.2 and 18.3 +/- 8.2 months, respectively. The primary end point of the study was survival free of major adverse cardiac events at 1 year. The secondary end points were survival free of the need for revascularization of the target lesion and definite stent thrombosis. The baseline clinical and angiographic parameters were comparable between the 2 groups. The freedom from major adverse cardiac event rate at 1 year of follow-up was 4.5% and 13.6% (p = 0.0663) for the EES and PES groups, respectively. The target lesion revascularization (TLR) rates were greater in the PES group at 1 year of follow-up compared to the EES group (1% vs 11.5%, p = 0.0193). The rate of myocardial infarction, death, and definite stent thrombosis for the EES and PES groups at 1 year of follow-up was 0% versus 4.2% (p = 0.0984), 3% versus 2.1% (p = 0.6855), and 0% versus 2.1% (p = 0.2382), respectively. The use of a PES for treatment of ISR was the only independent predictor of recurrent TLR at 1 year of follow-up (odds ratios 1.11, 95% confidence interval 1.05 to 1.18; p = 0.0193). During the complete follow-up period, the rates of TLR, myocardial infarction, death, major adverse cardiac events, and definite stent thrombosis were not different between the 2 treatment groups. In conclusion, EES resulted in reduced rates of TLR at 1 year of follow-up compared to PES when used for treatment of BMS ISR. However, at long-term follow-up, the event rates between EES and PES were comparable after treatment of BMS ISR. PMID- 21624554 TI - Meta-analysis of randomized trials focusing on prevention of the postpericardiotomy syndrome. AB - The natural history of postpericardiotomy syndrome (PPS), a relatively common complication of cardiac surgery, varies from mild self-limited episodes to cases with protracted courses, recurrences, and readmissions. Preventive strategies may be valuable to decrease morbidity and management costs. We thus aimed to conduct a comprehensive systematic review on available data for pharmacologic primary prevention of PPS. Controlled clinical studies were searched in several databases and were included provided they focused on pharmacologic primary prevention of PPS. Random-effect odds ratios (ORs) were computed for occurrence of PPS. From the initial sample of 343 citations, 4 controlled clinical trials for primary prevention of PPS were finally included (894 patients); 3 studies were double blind randomized controlled trials (RCTs). Treatment comparisons were colchicine versus placebo (2 RCTs enrolling 471 patients), methylprednisolone versus placebo (1 RCT on 246 pediatric patients), and aspirin versus historical controls (1 nonrandomized study on 177 pediatric patients). Meta-analytic pooling showed that colchicine was associated with decreased risk of PPS (OR 0.38, 0.22 to 0.65). Data on methylprednisolone (OR 1.13, 0.57 to 2.25) or aspirin (OR 1.00, 0.16 to 6.11) were negative but inconclusive because these were based on 1 study and/or a nonrandomized design. In conclusion, clinical evidence for primary prevention of PPS is still limited to few studies of variable quality. Nevertheless, available data suggest a beneficial profile for colchicine and open a new therapeutic strategy for prevention of PPS. PMID- 21624555 TI - Cost analysis of percutaneous pulmonary valve replacement. AB - Percutaneous pulmonary valve implantation is currently being used to treat right ventricular outflow tract obstruction or insufficiency in congenital heart disease. Presumably this alternative to surgical conduit replacement may result in cost savings owing to shorter hospital stays; however, a formal cost comparison has not been undertaken. Total hospital costs of percutaneous pulmonary valve implantation were compared to costs of surgical conduit replacement. Midterm cost-savings analysis was then modeled over 5 years using initial costs and reintervention rates. Need for surgical or transcatheter reintervention was derived from published data (5-year freedom from reintervention was assumed to be 53% for percutaneous pulmonary valves and 90% for surgical conduits). Cost of Melody valve and delivery device ($30,500) was higher than the conduit cost ($8,700), but total procedural costs were nearly identical at just less $50,000 for each procedure. When considering the increased need for reintervention in patients with Melody valves, surgical conduit revision results in moderate cost savings at 5 years after the initial procedure ($19,928 per patient). In conclusion, Melody valve implantation compares reasonably well to surgical conduit revision despite the added midterm costs, but ongoing analysis including the impact of nonsurgical options on quality-of-life measurements and improvement of reintervention rates for percutaneously placed valves needs to be considered. PMID- 21624556 TI - Clinical excellence in cardiology. AB - A recent study identified 7 domains of clinical excellence on the basis of interviews with "clinically excellent" physicians at academic institutions in the United States: (1) communication and interpersonal skills, (2) professionalism and humanism, (3) diagnostic acumen, (4) skillful negotiation of the health care system, (5) knowledge, (6) taking a scholarly approach to clinical practice, and (7) having passion for clinical medicine. What constitutes clinical excellence in cardiology has not previously been defined. The author discusses clinical excellence in cardiology using the framework of these 7 domains and also considers the additional domain of clinical experience. Specific aspects of the domains of clinical excellence that are of greatest relevance to cardiology are highlighted. In conclusion, this discussion characterizes what constitutes clinical excellence in cardiology and should stimulate additional discussion of the topic and an examination of how the domains of clinical excellence in cardiology are related to specific patient outcomes. PMID- 21624557 TI - Microbial keratitis after penetrating keratoplasty: impact of sutures. AB - PURPOSE: To determine the impact of presence or absence of sutures in cases with post-penetrating keratoplasty (PKP) microbial keratitis. DESIGN: A 10-year retrospective chart review of post-PKP patients admitted with microbial keratitis at the Royal Victorian Eye and Ear Hospital, Melbourne, between January 1998 and December 2008 was undertaken. METHODS: Patients were categorized in 2 groups, "sutures present" and "sutures absent." Main parameters evaluated were clinical and microbiological profile and treatment outcome. RESULTS: One hundred and twenty-two episodes of microbial keratitis were noted in 101 patients: 71 (58.2%) with sutures present and 51 (41.8%) with sutures absent. Overall, pseudophakic bullous keratopathy was the most common indication for keratoplasty (P=.92). Ocular surface disorder was the commonest risk factor associated with the occurrence of infection in both groups (P=.17). Infections caused by Moraxella sp. (P=.001) were significantly more common in the "sutures absent" group. Surgical interventions were required for 47 episodes (39%), with corneal gluing performed in significantly higher number of cases in the "sutures absent" group (40% vs 15%; P=.05). Multivariate analyses did not reveal any significant associations. Final mean visual acuity outcome was poorer in the "sutures absent" group (logMAR 2.10 +/- 0.92 vs 1.76 +/- 0.96; P=.04). CONCLUSIONS: Corneal graft infections, in the presence and absence of sutures, share similar indications and risk factors. However, infections caused by indolent microorganisms were more prevalent in grafts without sutures. This group of patients required a higher number of surgical interventions in the form of corneal gluing and the overall visual outcome was poor. PMID- 21624558 TI - Letter from the editors: positron emission tomography radiopharmaceuticals current status. PMID- 21624559 TI - Guest Editorial: new PET radiopharmaceuticals as molecular imaging probes. PMID- 21624560 TI - A broad overview of positron emission tomography radiopharmaceuticals and clinical applications: what is new? AB - Positron emission tomography (PET)/computed tomography (CT) is a rapidly expanding imaging modality, thanks to the availability of compact medical cyclotrons and automated chemistry synthesis modules for the production of PET radiopharmaceuticals. Despite the availability of many radiotracers, [(18)F]fluorodeoxyglucose (FDG) is currently the most widely used radiopharmaceutical in PET, and the field of molecular imaging is anxiously awaiting the introduction of new PET radiopharmaceuticals for routine clinical use. During the last five years, several proprietary PET radiopharmaceuticals have been developed by major companies, and these new agents are in different stages of clinical evaluation. These new PET drugs are designed for imaging brain beta amyloid, myocardial perfusion, amino acid transport, angiogenesis, and tumor antigen expression. In addition, the National Cancer Institute, Society of Nuclear Medicine Clinical Trials Network, and the American College of Radiology Imaging Network have been conducting multicenter clinical trials with several nonproprietary PET drugs such as sodium [(18)F]fluoride, [(18)F]fluorothymidine, [(18)F]fluoromisonidazole, and (64)Cu-labeled diacetyl-bis (N(4) methylthiosemicarbazone. All new PET radiopharmaceuticals, like any other drugs, must be manufactured under current good manufacturing practices as required by the Food and Drug Administration before clinical evaluation (phases I, II, and III) and submission of new drug application. This review briefly describes the chemistry, mechanisms(s) of localization, and clinical application of both proprietary and nonproprietary new PET drugs under multicenter clinical evaluation. PMID- 21624562 TI - Positron emission tomography radiopharmaceuticals for imaging brain Beta-amyloid. AB - Alzheimer's disease (AD) is defined histologically by the presence of extracellular beta-amyloid (Abeta) plaques and intraneuronal neurofibrillary tangles in the cerebral cortex. The diagnosis of dementia, along with the prediction of who will develop dementia, has been assisted by magnetic resonance imaging and positron emission tomography (PET) by using [(18)F]fluorodeoxyglucose (FDG). These techniques, however, are not specific for AD. Based on the chemistry of histologic staining dyes, several Abeta-specific positron-emitting radiotracers have been developed to image neuropathology of AD. Among these, [(11)C]PiB is the most studied Abeta-binding PET radiopharmaceutical in the world. The histologic and biochemical specificity of PiB binding across different regions of the AD brain was demonstrated by showing a direct correlation between Abeta-containing amyloid plaques and in vivo [(11)C]PiB retention measured by PET imaging. Because (11)C is not ideal for commercialization, several (18)F-labeled tracers have been developed. At this time, [(18)F]3'-F-PiB (Flutemetamol), (18)F AV-45 (Florbetapir), and (18)F-AV-1 (Florbetaben) are undergoing extensive phase II and III clinical trials. This article provides a brief review of the amyloid biology and chemistry of Abeta-specific (11)C and (18)F-PET radiopharmaceuticals. Clinical trials have clearly documented that PET radiopharmaceuticals capable of assessing Abeta content in vivo in the brains of AD subjects and subjects with mild cognitive impairment will be important as diagnostic agents to detect in vivo amyloid brain pathology. In addition, PET amyloid imaging will also help test the amyloid cascade hypothesis of AD and as an aid to assess the efficacy of antiamyloid therapeutics currently under development in clinical trials. PMID- 21624561 TI - The next generation of positron emission tomography radiopharmaceuticals in oncology. AB - Although (18)F-fluorodeoxyglucose ((18)F-FDG) is still the most widely used positron emission tomography (PET) radiotracer, there are a few well-known limitations to its use. The last decade has seen the development of new PET probes for in vivo visualization of specific molecular targets, along with important technical advances in the production of positron-emitting radionuclides and their related labeling methods. As such, a broad range of new PET tracers are in preclinical development or have recently entered clinical trials. The topics covered in this review include labeling methods, biological targets, and the most recent preclinical or clinical data of some of the next generation of PET radiopharmaceuticals. This review, which is by no means exhaustive, has been separated into sections related to the PET radionuclide used for radiolabeling: fluorine-18, for the labeling of agents such as FACBC, FDHT, choline, and Galacto RGD; carbon-11, for the labeling of choline; gallium-68, for the labeling of peptides such as DOTATOC and bombesin analogs; and the long-lived radionuclides iodine-124 and zirconium-89 for the labeling of monoclonal antibodies cG250, and J591 and trastuzumab, respectively. PMID- 21624563 TI - Florbetapir f-18: a histopathologically validated Beta-amyloid positron emission tomography imaging agent. AB - Florbetapir F-18 is a molecular imaging agent combining high affinity for beta amyloid, pharmacokinetic properties that allow positron emission tomography (PET) imaging within a convenient time after dose administration, and the wide availability of the radionuclide fluorine-18. Florbetapir F-18 is prepared by nucleophilic radiofluorination in approximately 60 minutes with a decay-corrected yield of 20%-40% and with a specific activity typically exceeding 100 Ci/mmol. The florbetapir F-18 dissociation constant (K(d)) for binding to beta-amyloid in brain tissue from Alzheimer's disease (AD) patients was 3.7 +/- 0.3 nmol/L, and the maximum binding capacity (B(max)) was 8800 +/- 1600 fmol/mg protein. Autoradiography studies have shown that florbetapir F-18 selectively binds to beta-amyloid aggregates in AD patient brain tissue, and the binding intensity is correlated with the density of beta-amyloid quantified by standard neuropathologic techniques. Studies in animals revealed no safety concerns and rapid and transient normal brain uptake (6.8% injected dose/g at 2 minutes and 1.9% injected dose/g at 60 minutes in the mouse). Florbetapir F-18 has been well tolerated in studies of more than 2000 human subjects. Biodistribution studies in humans revealed predominantly hepatobiliary excretion. The whole body effective dose was 7 mSv from a dose of 370 MBq. The pharmacokinetic of florbetapir F-18 make it possible to obtain a PET image with a brief (10 minutes) acquisition time within a convenient time window of 30-90 minutes after dose administration. Clinical studies have demonstrated a clear correlation between in vivo PET imaging with florbetapir F-18 and postmortem histopathologic quantitation of beta amyloid in the brain. PMID- 21624564 TI - The next generation of cardiac positron emission tomography imaging agents: discovery of flurpiridaz F-18 for detection of coronary disease. AB - Myocardial perfusion imaging (MPI) with thallium 201 ((201)Tl) or (99m)Tc based imaging agents has become a major tool for noninvasive identification of coronary artery disease (CAD). However, single photon emission computed tomography (SPECT) imaging with the current agents is vulnerable to artifacts associated with soft tissue attenuation, proximal gastrointestinal activity, image quality, and suboptimal sensitivity and is limited by the degree of first-pass myocardial extraction. The development of (18)F-based flurpiridaz F-18 takes advantage of positron emission tomography (PET) to overcome many of the imaging issues and structural design to achieve an ideal MPI agent profile. Flurpiridaz F-18 was designed to bind to mitochondrial complex I with high affinity and demonstrates high heart uptake in multiple species with clear delineation of perfusion deficits. It exhibits rapid uptake in the myocardium, prolonged retention, and superior extraction versus flow profiles compared with (201)Tl and (99m)Tc sestamibi. A first in man study has established the safety and dosimetry of flurpiridaz F-18 and confirmed high sustained cardiac uptake. Subsequent studies performed in CAD patients established the dose and timing needed to detect perfusion deficits when the agent is administered under rest and stress conditions. This review compares the current preclinical and clinical data with an ideal MPI agent profile. The assessment indicates flurpiridaz F-18 represents a new generation of PET MPI agents and demonstrates significantly improved molecular and imaging characteristics. PMID- 21624565 TI - (68)Ga-labeled DOTA-peptides and (68)Ga-labeled radiopharmaceuticals for positron emission tomography: current status of research, clinical applications, and future perspectives. AB - In this review we give an overview of current knowledge of (68)Ga-labeled pharmaceuticals, with focus on imaging receptor-mediated processes. A major advantage of a (68)Ge/(68)Ga generator is its continuous source of (68)Ga, independently from an on-site cyclotron. The increase in knowledge of purification and concentration of the eluate and the complex ligand chemistry has led to (68)Ga-labeled pharmaceuticals with major clinical impact. (68)Ga-labeled pharmaceuticals have the potential to cover all today's clinical options with (99m)Tc, with the concordant higher resolution of positron emission tomography (PET) in comparison with single photon emission computed tomography. (68)Ga labeled analogs of octreotide, such as DOTATOC, DOTANOC, and DOTA-TATE, are in clinical application in nuclear medicine, and these analogs are now the most frequently applied of all (68)Ga-labeled pharmaceuticals. All the above-mentioned items in favor of successful application of (68)Ga-labeled radiopharmaceuticals for imaging in patients are strong arguments for the development of a (68)Ge/(68)Ga generator with Marketing Authorization and thus to provide pharmaceutical grade eluate. Moreover, now not one United States Food and Drug Administration-approved or European Medicines Agency-approved (68)Ga radiopharmaceutical is available. As soon as these are achieved, a whole new radiopharmacy providing PET radiopharmaceuticals might develop. PMID- 21624566 TI - Subliminal presentation of other faces (but not own face) primes behavioral and evoked cortical processing of empathy for pain. AB - Current research on empathy for pain emphasizes the overlap in the neural response between the first-hand experience of pain and its perception in others. However, recent studies suggest that the perception of the pain of others may reflect the processing of a threat or negative arousal rather than an automatic pro-social response. It can thus be suggested that pain processing of other related, but not self-related, information could imply danger rather than empathy, due to the possible threat represented in the expressions of others (especially if associated with pain stimuli). To test this hypothesis, two experiments considering subliminal stimuli were designed. In Experiment 1, neutral and semantic pain expressions previously primed with own or other faces were presented to participants. When other-face priming was used, only the detection of semantic pain expressions was facilitated. In Experiment 2, pictures with pain and neutral scenarios previously used in ERP and fMRI research were used in a categorization task. Those pictures were primed with own or other faces following the same procedure as in Experiment 1 while ERPs were recorded. Early (N1) and late (P3) cortical responses between pain and no-pain were modulated only in the other-face priming condition. These results support the threat value of pain hypothesis and suggest the necessity for the inclusion of own- versus other-related information in future empathy for pain research. PMID- 21624567 TI - Lifespan aging and belief reasoning: Influences of executive function and social cue decoding. AB - Older adults often perform poorly on Theory of Mind (ToM) tests that require understanding of others' beliefs and intentions. The course and specificity of age changes in belief reasoning across the adult lifespan is unclear, as is the cause of the age effects. Cognitive and neuropsychological models predict that two types of processing might influence age differences in belief reasoning: executive functioning and social cue detection. In the current study we assessed 129 adults aged between 18 and 86 on novel measures of ToM (video clips and verbal vignettes), which manipulated whether true or false belief reasoning was required. On both video and verbal tasks, older adults (aged 65-88) had specific impairments in false belief reasoning, but showed no such problem in performing true belief tasks. Middle-aged adults (aged 40-64) generally performed as well as the younger adults (aged 18-39). Difficulties in updating information in working memory (but not inhibitory problems) partially mediated the age differences in false belief reasoning. Also, the ability to decode biological motion, indexing social cue detection, partially mediated age-related variance in the ability to interpret false beliefs. These results indicate that age differences in decoding social cues and updating information in memory may be important influences on the specific problems encountered when reasoning about false beliefs in old age. PMID- 21624568 TI - Sexual arousal and related concepts: an introduction. PMID- 21624569 TI - James Halperin's The truth machine: a fictional representation of neurologically based lie detection. PMID- 21624570 TI - Psychiatric classification in the setting of medical disease: comparing the clinical value of different proposals. PMID- 21624571 TI - Differing perspectives on diagnostic proposals for the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition. PMID- 21624572 TI - Chronic fatigue syndrome: neurological, mental or both. PMID- 21624573 TI - Chronic fatigue syndrome: labels, meanings and consequences. AB - In this month's issue, we report a survey of members of the Association of British Neurologists, which asked if they viewed chronic fatigue syndrome (CFS) as a neurological condition--84% of respondents did not. This is at odds with current classification in ICD-10. We discuss the difficulties of classifying CFS and myalgic encephalopmeylitis (ME), including historical and sociological factors, the pitfalls of the physical/psychological dichotomy and why classification matters to doctors and patients. PMID- 21624574 TI - Biomarkers in burnout: a systematic review. AB - BACKGROUND: Burnout is a stress state characterized by symptoms of mental exhaustion and physical fatigue, detachment from work, and feelings of diminished competence. Several biomarkers have been tested for association with burnout, but the results are conflicting. AIM: The objective of this review was to identify potential biomarkers for burnout. METHODS: We carried out a systematic review of studies comparing biomarkers in individuals with burnout and healthy controls, or individuals with low scores and those with high scores on burnout questionnaires. Literature searches in MEDLINE and EMBASE were performed. We describe biomarkers on which at least three studies were available. Where appropriate, a meta analysis was carried out. RESULTS: We identified 31 studies on 38 biomarkers involved in the hypothalamus-pituitary-adrenal axis, autonomic nervous system, immune system, metabolic processes, antioxidant defense, hormones, and sleep. At least 3 studies were available for cortisol in saliva and blood, blood pressure, heart rate, cholesterol, dehydroepiandrosterone sulfate, (numbers or activity of) natural killer cells, C-reactive protein, and prolactin. The comparability of studies was limited, due to differences in the methods used to characterize patients and controls, to assess biomarkers, and to control for confounders. Furthermore, burnout was operationalized in different ways. Meta-analyses showed no differences for cortisol awakening response and cortisol awakening response after administration of dexamethasone, cortisol in blood, and blood pressure. CONCLUSIONS: No potential biomarkers for burnout were found, largely due to the incomparability of studies. We emphasize the need for a dimensional and longitudinal approach in future research to account for the heterogeneity of burnout. PMID- 21624575 TI - Differential effects of alexithymia subscales on autonomic reactivity and anxiety during social stress. AB - OBJECTIVES: Alexithymia is characterized by a difficulty in identifying and describing one's emotions. Recent research has associated differential effects of the alexithymia facets to hypothalamic-pituitary-adrenal (HPA) axis markers during stress. This study aimed to analyze how the facets of alexithymia interact with autonomic reactivity as well as self- and observer-rated anxiety during a social stress task. METHODS: With the use of a public-speaking paradigm, skin conductance levels (SCLs) and heart rate (HR) during the defined periods of baseline, preparation, stress, and recovery were assessed in 60 volunteers (42 females, mean age 22.8) categorized as having either high (HDA) or low (LDA) degrees of alexithymia. RESULTS: We found smaller SCLs during preparation and speech in the HDA group. Regression analyses indicated that only the alexithymia facet "difficulty in describing feelings" (DDF) was associated with smaller electrodermal responses. In the HDA group, self- and observer-rated anxiety was higher in the HDA than in the LDA group, which was attributable to higher scores in the subscales "difficulty in identifying feelings" (DIF) and "externally oriented thinking" (EOT). CONCLUSIONS: Our data support and specify the decoupling hypothesis of alexithymia by showing that the facets of alexithymia are differentially related to autonomic reactivity as well as self- and observer rated anxiety during social stress. PMID- 21624576 TI - Goal conflict, distress, and pain in women with fibromyalgia: a daily diary study. AB - OBJECTIVES: A chronic illness such as fibromyalgia can interfere with daily activities and goals by limiting available resources, including time and energy. This leads to competition between goals, known as goal conflict. The purpose of this study was to determine if goal conflict increases symptoms in women with fibromyalgia and whether symptoms lead to perceptions of goal conflict. METHODS: Women with fibromyalgia (N=27) recorded their pain, emotional distress, and fatigue each morning and evening for five consecutive days. Each evening, they listed that day's goals, rating goals on their level of conflict. Goal conflict was also rated by independent raters, and a difference score reflected goal conflict discrepancy. RESULTS: On days with higher goal conflict, pain increased more from morning to evening (gamma=1.71, 95% confidence interval=0.32-3.09, P<.05). On days with higher morning emotional distress, goal conflict was overestimated (gamma=0.075, 95% confidence interval=0.035-0.116, P<.05). Women who had a higher symptom burden also typically overestimated their goal conflict relative to those with fewer symptoms (P<.05 for all). CONCLUSIONS: Goal pursuit may deplete psychological and physical resources in this vulnerable population, resulting in higher pain. Conversely, emotional distress may affect perception of goal conflict, resulting in less ambitious goal pursuit. Understanding the dynamic relationship between goal conflict and fibromyalgia symptoms may lead to more effective management of limited resources and pursuit of daily goals with fibromyalgia. PMID- 21624577 TI - Prevalence of and factors associated with psychiatric morbidity in chronic pain patients. AB - OBJECTIVE: The reported prevalence of psychiatric morbidity in chronic pain patients (CCPs) was high, although it varied tremendously since structured diagnostic instruments were seldom used for diagnosis in previous studies. Study in this area after the launching of the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) was scarce. This study serves to estimate the prevalence of psychiatric morbidity in patients attending a chronic pain clinic by using the Structured Clinical Interview for DSM-IV (SCID) Axis I disorders and to identify factors highly associated with psychiatric disorders, particularly depression, which is treatable. METHOD: Consecutive patients attending a chronic pain clinic were recruited during a 6-month period. Psychiatric diagnoses were made by using the SCID. Logistic regression was used to identify factors predicting overall psychiatric morbidity and depression. RESULTS: Prevalence of psychiatric disorders in this 89-patient sample was 62.9%. Current major depressive disorder was present in 31.5% and somatoform disorders in 33.7%. Anxiety disorders and current substance use disorders each constituted 18. "Younger age of onset of pain" (odds ratio [OR]=0.956, P<.05) and "higher pain intensity" (OR=1.544, P<.001) were independently associated with presence of psychiatric disorders. "Higher pain intensity" (OR=13.7, P<.05), "negative pain cognition" (OR=0.967, P<.05) and "problems with social and leisure activities" (OR=38.5, P<.05) were associated with depression. CONCLUSION: Prevalence of psychiatric disorders in this Chinese chronic pain clinic sample with reference to the DSM-IV was similar to that reported in previous studies. Specific factors were identified to alert pain physicians to underlying psychiatric disorders. PMID- 21624578 TI - Comorbid subjective health complaints in patients with sciatica: a prospective study including comparison with the general population. AB - OBJECTIVE: Chronic nonspecific low back pain is accompanied by high rates of comorbid mental and physical conditions. The aims of this study were to investigate if patients with specific back pain, that is, sciatica caused by lumbar herniation, report higher rates of subjective health complaints (SHCs) than the general population and if there is an association between change in sciatica symptoms and change in SHCs over a 12-month period. METHODS: A multicenter cohort study of 466 sciatica patients was conducted with follow-up at 3 months and 1 year. Comorbid SHCs were measured by 27 items of the SHC inventory. Odds ratios (ORs) for each SHC were calculated with comparison to a general population sample (n=928) by logistic regression. The SHC number was calculated by summing all complaints present. RESULTS: At baseline, the ORs for reporting SHCs for the sciatica patients were significantly elevated in 15 of the 27 items with a mean (S.D.) SHC number of 7.5 (4.4), compared to 5.2 (4.4) in the general population (P<.01). Among those who during the 1-year follow-up period fully recovered from their sciatica, the SHC number was reduced to normal levels. Among those with persisting or worsening sciatica, the number increased to a level almost double that of the general population. CONCLUSION: Compared to the general population, the prevalence of subjective health complaints in sciatica is increased. During follow-up, the number of health complaints increased in patients with persisting or worsening sciatica. PMID- 21624579 TI - Hyperventilation complaints in music performance anxiety among classical music students. AB - OBJECTIVE: Despite the importance of respiration and hyperventilation in anxiety disorders, research on breathing disturbances associated with hyperventilation is rare in the field of music performance anxiety (MPA, also known as stage fright). The only comparable study in this area reported a positive correlation between negative feelings of MPA and hyperventilation complaints during performance. The goals of this study were (a) to extend these previous findings to the period before performance, (b) to test whether a positive correlation also exists between hyperventilation complaints and the experience of stage fright as a problem, (c) to investigate instrument-specific symptom reporting, and (d) to confirm gender differences in negative feelings of MPA and hyperventilation complaints reported in other studies. METHODS: We assessed 169 university students of classical music with a questionnaire comprising: the State-Trait Anxiety Inventory for negative feelings of MPA, the Nijmegen Questionnaire for hyperventilation complaints, and a single item for the experience of stage fright as a problem. RESULTS: We found a significant positive correlation between hyperventilation complaints and negative feelings of MPA before performance and a significant positive correlation between hyperventilation complaints and the experience of stage fright as a problem. Wind musicians/singers reported a significantly higher frequency of respiratory symptoms than other musicians. Furthermore, women scored significantly higher on hyperventilation complaints and negative feelings of MPA. CONCLUSION: These results further the findings of previous reports by suggesting that breathing disturbances associated with hyperventilation may play a role in MPA prior to going on stage. Experimental studies are needed to confirm whether hyperventilation complaints associated with negative feelings of MPA manifest themselves at the physiological level. PMID- 21624580 TI - Advice on total-score reliability issues in psychosomatic measurement. AB - OBJECTIVE: This article addresses three reliability issues that are problematic in the construction of scales intended for use in psychosomatic research, illustrates how these problems may lead to errors, and suggests solutions. METHODS: We used psychometric results and present five computational studies. The first, third, and fourth studies are based on the generation of artificial data from psychometric models in combination with distributions for scale scores, as is common in psychometric research, whereas the second and fifth studies are analytical. RESULTS: The power of Student's t test depends more on sample size than on total-score reliability, but reliability must be high when one estimates correlations involving test scores. Short scales often do not allow total scores to be significantly different from a cutoff score. Coefficient alpha is uninformative about the factorial structure of questionnaires and is one of the weakest estimators of total-score reliability. CONCLUSIONS: The relationship between questionnaire length/reliability and statistical power is complex. Both in research and individual diagnostics, we recommend the use of highly reliable scales so as to reduce the chance of faulty decisions. The conclusion calls for profound statistical research producing hands-on rules for researchers to act upon. Factor analysis should be used to assess the internal consistency of questionnaires. As a reliability estimator, alpha should be replaced by better and readily available methods. PMID- 21624581 TI - Is chronic fatigue syndrome a neurological condition? A survey of UK neurologists. PMID- 21624582 TI - On the history of the European Association of Consultation-Liaison Psychiatry and Psychosomatics. PMID- 21624583 TI - Melena from jejunal mucosal varices caused by esophageal variceal sclerotherapy induced splenic arteriovenous fistula. PMID- 21624584 TI - Gastrocolic fistula related to NSAID-induced gastric ulcer. PMID- 21624585 TI - Editorial comment. PMID- 21624587 TI - Editorial comment. PMID- 21624588 TI - Editorial comment. PMID- 21624590 TI - Nocturia in interstitial cystitis/painful bladder syndrome. AB - OBJECTIVES: To explore the roles of pain and urgency in the nocturia of patients with interstitial cystitis/painful bladder syndrome (IC/PBS). METHODS: In a longitudinal study of incident IC/PBS cases, we assessed the associations of the presence and severity of nocturia with the presence and severity of pain and urgency, using multivariate analyses when necessary. Additionally, we simply asked patients with IC/PBS what awakens them at night. RESULTS: The multivariate analyses revealed associations of urgency with the presence and severity of nocturia and of bladder pain with the severity of nocturia. Direct queries of patients with IC/PBS about urgency and the reasons for awakening demonstrated that bladder pain might have played a twofold role: directly in awakening a large minority of patients and possibly indirectly in the majority by generating the sensation of urgency. CONCLUSIONS: These findings are consistent with urinary urgency and bladder pain each being in the causal pathway leading to nocturia in patients with IC/PBS. PMID- 21624591 TI - Editorial comment. PMID- 21624593 TI - Editorial comment. PMID- 21624594 TI - Editorial comment. PMID- 21624595 TI - Electroejaculatory stimulation and its implications for male infertility in spinal cord injury: a short history through four decades of sperm retrieval (1975 2010). PMID- 21624596 TI - Editorial comment. PMID- 21624597 TI - Editorial comment. PMID- 21624598 TI - Editorial comment. PMID- 21624599 TI - Editorial comment. PMID- 21624601 TI - Editorial comment. PMID- 21624602 TI - Editorial comment. PMID- 21624604 TI - Editorial comment. PMID- 21624606 TI - Editorial comment. PMID- 21624607 TI - Kindlin-2 expression in arsenite- and cadmium-transformed bladder cancer cell lines and in archival specimens of human bladder cancer. AB - OBJECTIVE: To confirm a microarray study that suggested that Kindlin-2 might play a role in the development and progression of bladder cancer. There has been no previous examination of Kindlin-2 expression in human bladder cancer. METHODS: A combination of real-time polymerase chain reaction, Western analysis, and immunohistochemistry was used to characterize Kindlin-2 expression in arsenite (As(+3))- and cadmium (Cd(+2))-transformed human cell lines, their tumor transplants in immunocompromised mice, and in archival specimens of human bladder and bladder cancer. RESULTS: The results show that the Kindlin-2 expression patterns in the cell lines were not duplicated in the tumor tissues. However, it was shown that Kindlin-2 was expressed in the stromal element of all the transplanted tumors and archival specimens of human bladder cancer. It was also shown that a small number of high-grade invasive urothelial cancers have focal expression of Kindlin-2 in the tumor cells. CONCLUSION: Kindlin-2 is expressed in the stromal component of most, if not all, human bladder cancers. Kindlin-2 is not expressed in normal urothelium. Kindlin-2 is expressed in a small subset of high-grade invasive bladder cancers and may have potential as a prognostic marker for tumor progression. PMID- 21624608 TI - Effect of the phytotherapeutic agent Eviprostat on inflammatory changes and cytokine production in a rat model of nonbacterial prostatitis. AB - OBJECTIVES: To examine the effect of the phytotherapeutic agent Eviprostat on the stromal-to-epithelial (S/E) ratio, level of macrophage infiltration, expression of the macrophage inhibitory cytokine-1 (Mic1) gene, and tumor necrosis factor alpha (TNF-alpha) and interleukin-8 (IL-8) concentrations in prostate tissues in a rat model of nonbacterial prostatitis (NBP). MATERIALS AND METHODS: Ten-month old Wistar rats were divided into 4 groups of 10: (1) NBP non-mixed feed (prostatitis control group); (2) NBP Eviprostat (0.1%) mixed feed (prostatitis Eviprostat group); (3) non-NBP non-mixed feed (nonprostatitis control group); and (4) non-NBP Eviprostat mixed-feed (nonprostatitis Eviprostat group). NBP was induced by castration followed by daily subcutaneous injection of 17beta estradiol for 30 days. Ventral prostate lobes were histopathologically examined with Masson's trichrome staining or immunostaining with antimacrophage antibody. Mic1 mRNA levels were quantified by real-time reverse transcriptase polymerase chain reaction. Tissue concentrations of TNF-alpha and IL-8 were determined by enzyme-linked immunosorbent assay. RESULTS: Stroma was the most abundant in prostatitis control rats. The mean S/E ratio in prostatitis Eviprostat rats was significantly lower than in prostatitis control rats (P < .0001). The high levels of macrophage infiltration found in prostatitis control rats were significantly reduced in prostatitis Eviprostat rats (P < .0001). The up-regulation of the Mic1 gene observed in prostatitis control rat prostates was significantly suppressed in prostatitis Eviprostat rats (P < .0001). A marked suppression of TNF-alpha and IL-8 secretion was also observed in prostatitis Eviprostat rats (P < .05). CONCLUSIONS: Eviprostat significantly suppressed the S/E ratio, level of macrophage infiltration, Mic1 gene expression, and proinflammatory cytokines/chemokines in the prostate in a rat NBP model. PMID- 21624609 TI - Lovastatin causes diminished PSA secretion by inhibiting AR expression and function in LNCaP prostate cancer cells. AB - OBJECTIVES: To investigate why statin users display a noticeable decline in prostate specific antigen (PSA) as revealed in recent clinical trials, we tested the effects of lovastatin on the androgen signaling cascade in lymph node carcinoma of the prostate (LNCaP) prostate cancer cells. METHODS: Effects of lovastatin alone or in combination with a small interference RNA to inhibit AR expression on cell proliferation and induction of apoptosis were assessed by [(3)H] thymidine incorporation and caspase-3 activity assay. PSA levels were measured in the cell culture supernatant by immunoassay. In addition, expression and activity of AR and Akt/protein kinase B (Akt) were determined by Western blotting and real-time polymerase chain reaction as well as by luciferase reporter gene assay. RESULTS: Our results show that lovastatin significantly reduces AR expression and activity, resulting in decreased PSA levels. These effects were associated with inhibition of cell proliferation and induction of apoptosis. In addition, we observed that the Akt signaling pathway plays a pivotal role in lovastatin-mediated regulation of AR signaling. CONCLUSIONS: Our data suggest that the regular use of statins may have beneficial effects in statin users by preventing prostate cancer growth through inhibition of androgen activation and expression, resulting in diminished PSA levels. PMID- 21624611 TI - Re: Endo et al.: Anteroposterior dissection HoLEP: a modification to prevent transient stress urinary incontinence (Urology 2010;76:1451-1456). PMID- 21624612 TI - Re: Tollefson et al.: Serum calcium is not predictive of aggressive prostate cancer after radical prostatectomy (Urology 2011;77:1161-1165). PMID- 21624614 TI - Does sensitivity to criticism mediate the relationship between theory of mind and academic achievement? AB - This study adds to the growing research on school outcomes associated with individual differences in preschoolers' theory of mind skills by considering whether "costs" of theory of mind (e.g., sensitivity to criticism) actually help to foster children's academic achievement. A group of 60 Italian children was tested during the last year of preschool (Time 1, mean age=5years 6months) and in their first year (Time 2, mean age=6years 5months) and second year (Time 3, mean age=7years 5months) of primary school. Children's theory of mind, verbal ability, and social skills were evaluated at each time point. In addition, children's sensitivity to teacher criticism was assessed at Time 2 and teachers' ratings of children's academic achievement were gathered at Time 3. Mediation analyses showed that, independent of verbal ability and social skills, sensitivity to criticism at Time 2 mediated the association between theory of mind at Time 1 and academic achievement at Time 3. These findings highlight the importance of investigating the educational consequences of individual differences in preschoolers' theory of mind. PMID- 21624615 TI - Gene therapy for primary adaptive immune deficiencies. AB - Gene therapy has become an option for the treatment of 2 forms of severe combined immunodeficiency (SCID): X-linked SCID and adenosine deaminase deficiency. The results of clinical trials initiated more than 10 years ago testify to sustained and reproducible correction of the underlying T-cell immunodeficiency. Successful treatment is based on the selective advantage conferred on T-cell precursors through their expression of the therapeutic transgene. However, "first generation" retroviral vectors also caused leukemia in some patients with X linked SCID because of the constructs' tendency to insert into active genes (eg, proto-oncogenes) in progenitor cells and transactivate an oncogene through a viral element in the long terminal repeat. These elements have been deleted from the vectors now in use. Together with the use of lentiviral vectors (which are more potent for transducing stem cells), these advances should provide a basis for the safe and effective extension of gene therapy's indications in the field of primary immunodeficiencies. Nevertheless, this extension will have to be proved by examining the results of the ongoing clinical trials. PMID- 21624616 TI - Neonatal screening for severe combined immunodeficiency caused by an adenosine deaminase defect: a reliable and inexpensive method using tandem mass spectrometry. AB - BACKGROUND: Adenosine deaminase (ADA)-severe combined immunodeficiency (SCID) is an SCID caused by a defect in the enzyme adenosine deaminase. It is usually fatal in infancy because of severe recurrent infections. When diagnosis is made, permanent damage caused by infections or by metabolites is often present. Gene therapy, bone marrow transplantation, or enzyme therapy might be effective if performed early. ADA-SCID complies with all the criteria for inclusion in a newborn screening program. However, screening methods are still expensive or provide a non-negligible number of indeterminate results. OBJECTIVE: The aim of the present study was to develop a simple, reliable, and inexpensive method for diagnosis of ADA-SCID by using dried blood spot (DBS) samples taken at birth. Cost per test was calculated, including the cost for reagents, equipment, and operators. METHODS: DBS samples from 4 patients with genetically confirmed ADA SCID and 12,020 DBS samples from healthy newborns were examined. Adenosine and 2' deoxyadenosine were tested by using tandem mass spectrometry (PCT EP2010/070517). RESULTS: The mean levels of adenosine and 2'-deoxyadenosine were 7.8 +/- 3.1 and 8.5 +/- 6.0 MUmol/L, respectively, in affected children; adenosine was found at 0.23 +/- 0.09 MUmol/L, whereas 2'-deoxyadenosine was never detected in healthy control subjects (adenosine: P < 10(-6) [95% confidence limit, 7.59-7.78] and 2' deoxyadenosine: P < 10(-6) [95% confidence limit, 8.65-8.82] for control subjects vs patients with ADA-SCID). No indeterminate or false-positive results were found. Cost per test was ?0.01 ($0.013). A pilot population-based newborn screening for ADA-SCID has started in Tuscany, Italy. CONCLUSION: Tandem mass spectrometry can be used for diagnosis of one of the most frequent form of SCID at a negligible cost. PMID- 21624618 TI - Obesity and asthma: an association modified by age of asthma onset. AB - BACKGROUND: Studies of asthma phenotypes have identified obesity as a component of a group characterized by a high proportion of subjects with adult-onset asthma. However, whether age of asthma onset modifies the association between obesity and asthma is unknown. OBJECTIVES: We sought to compare the associations between body mass index (BMI) categories with physiological, inflammatory, and clinical parameters across age of asthma onset phenotypes; and to compare the rate of BMI change in relation to asthma duration, by age of onset asthma phenotypes. METHODS: From the Severe Asthma Research Program, we defined age of asthma onset as early (<12 years of age) and late (>=12 years of age). Comparisons of BMI categories were done within age-of-onset groups, and obesity was also compared across these groups. Multivariable logistic regression analysis was done to evaluate the association between BMI categories with health care use and respiratory symptoms and multivariable linear regression for the association between duration of asthma and weight gain (BMI change per year). An interaction between obesity and age of asthma onset was included in the multivariable analyses. RESULTS: The study population consisted of 1049 subjects, and the median age for asthma onset was 10 years (interquartile range, 4-25 years); 48% had late-onset asthma (>=12 years of age), and 52% had early-onset asthma (<12 years of age). Compared with obese subjects with late-onset asthma, obese subjects with early-onset asthma had more airway obstruction, bronchial hyperresponsiveness, and higher odds ratios of ever having 3 or more previous oral steroid tapers per year or intensive care unit admissions for asthma per preceding year (interactions between obesity and age of asthma onset were P = .055 and P = .02, respectively). In subjects with early-onset asthma but not in subjects with late-onset asthma, there was a significant association between increasing BMI and duration of asthma after adjusting for confounders. The interaction between asthma duration and age of asthma onset was a P value of less than .01. CONCLUSION: Asthmatic subjects are differentially affected by obesity based on whether they had asthma early (<12 years of age) or later in life. These results highlight the need to understand obesity as a comorbidity that affects specific clinical phenotypes and not all asthma subjects alike. PMID- 21624619 TI - Distinct immune effector pathways contribute to the full expression of peanut induced anaphylactic reactions in mice. AB - BACKGROUND: Food-induced anaphylaxis is often a severe allergic reaction characterized by multiorgan dysfunction and a potentially fatal outcome. OBJECTIVES: We sought to investigate the relative contribution of immunoglobulin dependent effector pathways to anaphylactic responses to food (ie, peanut). METHODS: Wild-type and various mutant mice were sensitized with peanut protein and cholera toxin by means of oral gavage weekly for 4 weeks. Mice were subjected to different cellular depletion and Fc receptor blocking strategies before challenge with peanut 1 week after the last sensitization. RESULTS: Our data indicate that pathways other than the classical mast cell (MC)-IgE pathway contribute to the full spectrum of anaphylactic reactions to peanut. We show that the single deletion of MCs, basophils, or phagocytes (ie, macrophages) prevents the most significant clinical outcome: death. Remarkably, the combined deficiency of MCs and phagocytes, but not MCs and basophils, averted nearly all clinical and physiological signs of anaphylaxis. Furthermore, blockade of both IgE and IgG1 signaling was necessary to abolish anaphylactic responses to peanut. Although MC responses occurred through IgE and IgG1, phagocyte responses were fully mediated through IgG1. CONCLUSIONS: Peanut-induced anaphylaxis is a process that involves the concerted action of multiple immune effector pathways, and thus interventions targeting a single pathway (eg, MC-IgE) might not be sufficient to fully prevent anaphylactic responses. PMID- 21624620 TI - Claudin-1 expression in airway smooth muscle exacerbates airway remodeling in asthmatic subjects. AB - BACKGROUND: Increased airway smooth muscle (ASM) mass is an essential component of airway remodeling and asthma development, and there is no medication specifically against it. Tight junction (TJ) proteins, which are expressed in endothelial and epithelial cells and affect tissue integrity, might exist in other types of cells and display additional functions in the asthmatic lung. OBJECTIVE: The aim of this study was to investigate the existence, regulation, and function of TJ proteins in ASM in asthmatic patients. METHODS: The expression and function of TJ proteins in primary ASM cell lines, human bronchial biopsy specimens, and a murine model of asthma were analyzed by means of RT-PCR, multispectral imaging flow cytometry, immunohistochemistry, Western blotting, 5 (and-6)-carboxyfluorescein diacetate succinimidyl ester staining, tritiated thymidine incorporation, wound-healing assay, and luminometric bead array. RESULTS: Increased claudin-1 expression was observed in ASM of asthmatic patients, as well as in a murine model of asthma-like airway inflammation. Whereas IL-1beta and TNF-alpha upregulated claudin-1 expression, it was downregulated by the T(H)2 cytokines IL-4 and IL-13 in primary human ASM cells. Claudin-1 was localized to the nucleus and cytoplasm but not to the cell surface in ASM cells. Claudin-1 played a central role in ASM cell proliferation, as demonstrated by increased ASM cell proliferation seen with overexpression and decreased proliferation seen with small interfering RNA knockdown of claudin-1. Overexpression of claudin-1 induced vascular endothelial growth factor and downregulated IL-6, IL-8, and IFN-gamma-induced protein 10 production by ASM cells. Claudin-1 upregulation by IL-1beta or TNF-alpha was suppressed by dexamethasone but not by rapamycin, FK506, or salbutamol. CONCLUSION: These results demonstrate that claudin-1 might play a role in airway remodeling in asthmatic patients by means of regulation of ASM cell proliferation, angiogenesis, and inflammation. PMID- 21624622 TI - Histopathologic evaluation of basilar artery atherosclerosis. AB - INTRODUCTION: There has been limited attention to pathological features of basilar artery atherosclerosis. It has been assumed that pathology of basilar artery atherosclerosis mimics that of other vascular beds. METHODS: To define the nature of the basilar artery atherosclerotic lesions, we analyzed postmortem intracranial artery samples from eight subjects with history of stroke. RESULTS: Atherosclerotic lesions were present in 7/8 arteries examined, with a mean estimated stenosis of 34%. Lumen thrombus with a disrupted fibrous cap was seen in 1 lesion; the remaining 6 lesions revealed a thick fibrous cap. Neovascularity and calcification were seen in 1 lesion and mild to moderate inflammation was seen in 3 lesions. Necrotic core was present in 4/7 lesions, and was associated with plaque rupture in the only disrupted lesion. CONCLUSIONS: Basilar artery atherosclerotic lesions were relatively benign in this series of patients presenting with stroke. While confirmation is needed with larger sample size, the relative paucity of neovascularity suggests a possibly distinctive histopathological profile. PMID- 21624623 TI - Admission hyperglycemia and serial infarct volume after t-PA therapy in patients with and without early recanalization. AB - BACKGROUND AND PURPOSE: The present study examined the effects of admission hyperglycemia and early recanalization (ER) after t-PA administration on infarct volume and patient outcome. METHODS: Acute ischemic stroke patients with major artery occlusion treated with t-PA within 3h of onset were studied prospectively. Hyperglycemia was identified as admitting blood glucose value>=130 mg/dl. We compared serial infarct volume and patient outcome between normoglycemic and hyperglycemic groups, and assessed correlation between admitting blood glucose value and ?infarct volume (7 days-baseline) between patients with and without ER. RESULTS: 97 patients (ICA occlusion in 30, M1 in 44, and M2 in 23 patients) were enrolled in the present study; 52 had hyperglycemia, and 40 had ER. The initial infarct volume did not differ between the normoglycemic and hyperglycemic groups. However, infarct volume at 7 days was larger in the hyperglycemic group than in the normoglycemic group (156.2+/-157.1cm(3), vs. 85.4+/-140.7 cm(3), P=0.0061) and the baseline admitting blood glucose value was correlated with Deltainfarct volume (7 days-baseline) (r=0.340, P=0.0014). Regarding ER, Deltainfarct volume (7 days-baseline) in patients without ER was correlated with admitting blood glucose value(r=0.372, P=0.0078). However, in patients with ER, Deltainfarct volume was not associated with admitting blood glucose value (r=0.225, P=0.1173). Good outcome (mRS 0-2) at 3 months was more frequent in normoglycemic patients than hyperglycemic patients (43.2% vs. 22.2%, P=0.0418). CONCLUSION: Admission hyperglycemia was associated with infarct volume expansion and patient outcome in t-PA patients. However, if ER occurs, hyperglycemia should not adversely affect infarct volume. PMID- 21624624 TI - Shape analysis of subcortical nuclei in Huntington's disease, global versus local atrophy--results from the TRACK-HD study. AB - Huntington's disease (HD) is characterized by brain atrophy. Localized atrophy of a specific structure could potentially be a more sensitive biomarker reflecting neuropathologic changes rather than global volume variation. We examined 90 TRACK HD participants of which 30 were premanifest HD, 30 were manifest HD and 30 were controls. Using FMRIB's Integrated Registration and Segmentation Tool, segmentations were obtained for the pallidum, caudate nucleus, putamen, thalamus, accumbens nucleus, amygdala, and hippocampus and overall volumes were calculated. A point distribution model of each structure was obtained using Growing and Adaptive Meshes. Permutation testing between groups was performed to detect local displacement in shape between groups. In premanifest HD overall volume loss occurred in the putamen, accumbens and caudate nucleus. Overall volume reductions in manifest HD were found in all subcortical structures, except the amygdala, as compared to controls. In premanifest HD shape analysis showed small areas of displacement in the putamen, pallidum, accumbens and caudate nucleus. When the premanifest group was split into two groups according to predicted disease onset, the premanifest HD group close to expected disease onset showed more pronounced displacements in caudate nucleus and putamen compared to premanifest HD far from disease onset or the total premanifest group. Analysis of shape in manifest HD showed widespread shape differences, most prominently in the caudal part of the accumbens nucleus, body of the caudate nucleus, putamen and dorsal part of the pallidum. We conclude that shape analysis provides new insights in localized intrastructural atrophy patterns in HD, but can also potentially serve as specific target areas for disease tracking. PMID- 21624625 TI - On the spectrum of acute dengue virus myositis. PMID- 21624626 TI - Structural health monitoring using polymer-based capacitive micromachined ultrasonic transducers (CMUTs). AB - Transducers based on a capacitive micromachined ultrasonic transducer (CMUT) design have been fabricated using a rapid prototyping technique. This results in a device that is constructed principally from polymers, in a process which is simple and inexpensive. The resultant devices can be attached to the surfaces of solids. Their peak sensitivity is in the 80-100 kHz range, making them ideal for applications such as acoustic emission and structural health monitoring. Good low frequency sensitivity leads to applications in vibration monitoring. PMID- 21624627 TI - The effect of UV/H2O2 treatment on disinfection by-product formation potential under simulated distribution system conditions. AB - Advanced oxidation with ultraviolet light and hydrogen peroxide (UV/H(2)O(2)) produces hydroxyl radicals that have the potential to degrade a wide-range of organic micro-pollutants in water. Yet, when this technology is used to reduce target contaminants, natural organic matter can be altered. This study evaluated disinfection by-product (DBP) precursor formation for UV/H(2)O(2) while reducing trace organic contaminants in natural water (>90% for target pharmaceuticals, pesticides and taste and odor producing compounds and 80% atrazine degradation). A year-long UV/H(2)O(2) pilot study was conducted to evaluate DBP precursor formation with varying water quality. The UV pilot reactors were operated to consistently achieve 80% atrazine degradation, allowing comparison of low pressure (LP) and medium pressure (MP) lamp technologies for DBP precursor formation. Two process waters of differing quality were used as pilot influent, i.e., before and after granular activated carbon adsorption. DBP precursors increased under most of the conditions studied. Regulated trihalomethane formation potential increased through the UV/H(2)O(2) reactors from 20 to 118%, depending on temperature and water quality. When Post-GAC water served as reactor influent, less DBPs were produced in comparison to conventionally treated water. Haloacetic acid (HAA5) increased when conventionally treated water served as UV/H(2)O(2) pilot influent, but only increased slightly (MP lamp) when GAC treated water served as pilot influent. No difference in 3-day simulated distribution system DBP concentration was observed between LP and MP UV reactors when 80% atrazine degradation was targeted. PMID- 21624628 TI - Oxygen consumption by a sediment bed for stagnant water: comparison to SOD with fluid flow. AB - A model of sedimentary oxygen demand (SOD) for stagnant water in a lake or a reservoir is presented. For the purposes of this paper, stagnant water is defined as the bottom layer of stratified water columns in relatively unproductive systems that are underlain by silt and sand-dominated sediments with low-organic carbon (C) and nitrogen (N). The modeling results are compared to those with fluid flow to investigate how flow over the sediment surface raises SOD compared to stagnant water, depending on flow velocity and biochemical activity in the sediment. SOD is found to be substantially limited by oxygen transfer in the water column when water is stagnant. When flow over the sediment surface is present, SOD becomes larger than that for stagnant water, depending on flow velocity and the biochemical oxygen uptake rate in the sediment. Flow over the sediment surface causes an insignificant raise in SOD when the biochemical oxygen uptake rate is small. The difference between SOD with fluid flow and SOD for stagnant water becomes significant as the biochemical oxygen uptake rate becomes larger, i.e. SOD is 10-100 times larger when flow over the sediment surface is present. PMID- 21624629 TI - Influences of thermal decontamination on mercury removal, soil properties, and repartitioning of coexisting heavy metals. AB - Thermal treatment is a useful tool to remove Hg from contaminated soils. However, thermal treatment may greatly alter the soil properties and cause the coexisting contaminants, especially trace metals, to transform and repartition. The metal repartitioning may increase the difficulty in the subsequent process of a treatment train approach. In this study, three Hg-contaminated soils were thermally treated to evaluate the effects of treating temperature and duration on Hg removal. Thermogravimetric analysis was performed to project the suitable heating parameters for subsequent bench-scale fixed-bed operation. Results showed that thermal decontamination at temperature>400 degrees C successfully lowered the Hg content to<20 mg kg(-1). The organic carbon content decreased by 0.06 0.11% and the change in soil particle size was less significant, even when the soils were thermally treated to 550 degrees C. Soil clay minerals such as kaolinite were shown to be decomposed. Aggregates were observed on the surface of soil particles after the treatment. The heavy metals tended to transform into acid-extractable, organic-matter bound, and residual forms from the Fe/Mn oxide bound form. These results suggest that thermal treatment may markedly influence the effectiveness of subsequent decontamination methods, such as acid washing or solvent extraction. PMID- 21624630 TI - Exposure to hazardous air pollutants along Oba Akran road, Lagos-Nigeria. AB - We measured toxic air pollutants along Oba Akran road in Lagos to evaluate pedestrian exposure. PM10, CO, O3, NO2, SO2, CH4, noise, wind velocity and temperature were measured simultaneously with portable analyzers. Our results showed that pedestrian exposure to PM10 (with an average of 274.6 MUg m(-3) for all samples) and CO (with an average of 19.27 ppm for all samples) was relatively high. CO is a traffic-related pollutant, so the influence of the local traffic emissions on CO levels is strong. The high concentration of the PM10 measured at the three environments also suggests that the traffic is a major source of ultrafine particles. The overall average concentrations for the 72-day experimental period for SO2, NO2 and O3 are 101.2, 62.5 and 0.32 ppb respectively, all of which are below the US national ambient air quality standards. Strong traffic impacts can be observed from the concentrations of some of these pollutants measured in these three environments. Most clear is a reflection of diesel truck traffic activity rich in black carbon concentrations. The diurnal variation of O3 and NO2 also showed that NO2 was depleted by photochemically formed O3 during the day and replenished at night as O3 was destroyed. A multivariate statistical analysis (Principal Component Analysis, Factor Analysis) has been applied to a set of data in order to determine the contribution of different sources. It was found that the main principal components, extracted from the air pollution data, were related to gasoline combustion, oil combustion and ozone interactions. PMID- 21624631 TI - Is the reproduction of Donax trunculus affected by their sites of origin contrasted by their level of contamination? AB - The reproductive cycle of bivalves is regulated by several natural environmental factors but exposure to chemical pollutants can also interfere and may result in advanced or delayed spawning season. To our knowledge, the gametogenic cycle of the suspension-feeder bivalve Donax trunculus has not yet been used as biomonitoring tool in ecotoxicological surveys. The aim of this study was to examine over a year physiological reproductive endpoints (sex-ratio, gametogenic and energy reserve cycles) and biological indices (condition index, allometry) in D. trunculus originating from two sites differing by their level of contamination. Specimens were collected bimonthly from November 2008 to October 2009 from a polluted site (Rades Meliane) and a comparatively reference site (Sidi Jehmi) in the Gulf of Tunis (Tunisia). Five stages were depicted by histological examination of gonads: undifferentiated, developing, mature, spawn and spent. Differences in the gametogenic cycle according to the site of origin of bivalves were observed. The spawning period began in March and was maximum in May in bivalves from both sites, but the percentage of spawning animals was higher in the polluted site vs the reference site. The spawning period was shorter in animals from the polluted site comparatively to the reference site. Energy reserves (glycogen, lipids) were higher in March and May comparatively to the other studied periods in bivalves from both sites. Lower energy reserves levels were usually observed in animals from the polluted site compared to the reference site. Seasonal variations of the condition index were associated to the reproductive and nutritive status of bivalves. Differences in allometry were depicted between bivalves from both studied sites. If we try to link allometry, energy reserves and reproduction, it can be hypothesized that for bivalves from the reference site, energy reserves are allocated to gametogenesis and length growth. For bivalves from the polluted site, energy reserves could be devoted to tolerance to chemical stress and to reproductive processes. Therefore, D. trunculus appears as a suitable sentinel species for the assessment of the ecotoxicological risk of contaminants such as endocrine disruptors. PMID- 21624632 TI - Phototransformation of ibuprofen and ketoprofen in aqueous solutions. AB - The UV (254 nm) and UV/VUV (254/185 nm) photolysis of two anti-inflammatory drugs, ibuprofen and ketoprofen, have been studied in aqueous solutions as a possible process for the removal of non-biodegradable compounds. We have examined the effects of dissolved oxygen and initial target concentration. Upon irradiation at 254 nm, the decomposition rate of ketoprofen is almost forty times higher as it of ibuprofen whilst VUV irradiation only increased the ibuprofen decomposition rate. The presence of dissolved oxygen accelerated the photodegradation of ibuprofen, whereas no effect was observed on the degradation of ketoprofen. The maximum quantum yield for the phototransformation was 0.2. The rate of mineralization in both cases was ~60%, even after 1h of treatment and this suggests the formation of stable by-products which were identified using GC MS and HPLC-MS, respectively. PMID- 21624633 TI - Concentrations, distribution, and bioaccumulation of synthetic musks in the Haihe River of China. AB - Seven typical synthetic musks (SMs) in the samples from the surface water, sediment and fish of the Haihe River were measured. The SM concentrations in the sediment and surface water of the Haihe River were significantly lower than those in the Dagu Drainage River and Chentaizi Drainage River (p<0.05). Along the flow direction, the SM concentrations in surface water and sediment tended to increase from the upstream to the downstream of Dagu Drainage River. The Bioaccumulation factors (BAFs) of galaxolide (HHCB) and tonalide (AHTN) were calculated at high levels in the muscles of crucian carp, common carp, and silver carp. Most of the biota-sediment accumulation factors (BSAFs) for HHCB and AHTN were higher than 1.7, suggesting magnification possibly exist in the musk bioaccumulations of the three fishes in the Haihe River. No significant differences in HHCB/AHTN ratios were observed among the water, fish, and sediment samples (p>0.05). However, the HHCB/AHTN values in the Haihe River were much lower than those in the Dagu Drainage River and Chentaizi Drainage River (p<0.05). Compared with several typical persistent organic pollutants (POPs), the musk concentrations were higher or comparable in the Haihe River. PMID- 21624634 TI - Emissions of unintentional persistent organic pollutants from open burning of municipal solid waste from developing countries. AB - Open burning of waste is the most significant source of polychlorinated dibenzo para-dioxins and dibenzofurans (PCDD/PCDF) in many national inventories prepared pursuant to the Stockholm Convention on Persistent Organic Pollutants. This is particularly true for developing countries. Emission factors for POPs such as PCDD/PCDF, dioxin-like polychlorinated biphenyls (dl-PCB) and penta- and hexachlorobenzenes (PeCBz/HCB) from open burning of municipal solid waste in China and Mexico are reported herein. Six different waste sources were studied varying from urban-industrial to semi-urban to rural. For PCDD/PCDF, the emission factors to air ranged from 3.0 to 650 ng TEQ kg(-1)waste and for dl-PCB from 0.092 to 54 ng TEQ kg(-1)waste. Emission factors for PeCBz (17-1200 ng kg( 1)waste) and HCB (24-1300 ng kg(-1)waste) spanned a wide but similar range. Within the datasets there is no indication of significant waste composition effect on emission factor with the exception of significantly higher Mexico rural samples. PMID- 21624635 TI - Do mobile phones of patients, companions and visitors carry multidrug-resistant hospital pathogens? AB - A cross-sectional study was conducted to determine bacterial colonization on the mobile phones (MPs) used by patients, patients' companions, visitors, and health care workers (HCWs). Significantly higher rates of pathogens (39.6% vs 20.6%, respectively; P = .02) were found in MPs of patients' (n = 48) versus the HCWs' (n = 12). There were also more multidrug pathogens in the patents' MPs including methicillin-resistant Staphylococcus aureus, extended-spectrum beta-lactamase producing Escherichia coli, and Klebsiella spp, high-level aminoglycoside resistant Enterococcus spp, and carabepenem-resistant Acinetobacter baumanii. Our findings suggest that mobile phones of patients, patients' companions, and visitors represent higher risk for nosocomial pathogen colonization than those of HCWs. Specific infection control measures may be required for this threat. PMID- 21624636 TI - Health care-associated infections studies project: an American Journal of Infection Control and National Healthcare Safety Network data quality collaboration case study 5. PMID- 21624637 TI - Randomized double-blind crossover trial of ultraviolet light-sanitized keyboards in a pediatric hospital. AB - The effectiveness of ultraviolet light disinfection of keyboards was assessed in the intensive care unit and emergency department of a pediatric hospital. Ultraviolet light disinfection was 67% effective (95% confidence interval, 46% 87%) in eliminating bacterial contamination as measured by quantitative bacterial culture. PMID- 21624638 TI - Comparison of dental measurement systems for taxonomic assignment of Neanderthal and modern human lower second deciduous molars. AB - Traditional morphometric approaches for taxonomic assignment of Neanderthal and modern human dental remains are mainly characterized by caliper measurements of tooth crowns. Several studies have recently described differences in dental tissue proportions and enamel thickness between Neanderthal and modern human teeth. At least for the lower second deciduous molar (dm(2)), a three-dimensional lateral relative enamel thickness index has been proposed for separating the two taxa. This index has the advantage over other measurements of being applicable to worn teeth because it ignores the occlusal aspect of the crown. Nevertheless, a comparative evaluation of traditional crown dimensions and lateral dental tissue proportion measurements for taxonomic assignment of Neanderthal and modern human dm(2)s has not yet been performed. In this study, we compare various parameters gathered from the lateral aspects of the crown. These parameters include crown diameters, height of the lateral wall of the crown (lateral crown height = LCH), lateral enamel thickness, and dentine volume of the lateral wall, including the volume of the coronal pulp chamber (lateral dentine plus pulp volume = LDPV), in a 3D digital sample of Neanderthal and modern human dm(2)s to evaluate their utility in separating the two taxa. The LDPV and the LCH allow us to discriminate between Neanderthals and modern humans with 88.5% and 92.3% accuracy, respectively. Though our results confirm that Neanderthal dm(2)s have lower relative enamel thickness (RET) index compared with modern humans (p = 0.005), only 70% of the specimens were correctly classified on the basis of the RET index. We also emphasize that results of the lateral enamel thickness method depend on the magnitude of the interproximal wear. Accordingly, we suggest using the LCH or the LDPV to discriminate between Neanderthal and modern human dm(2)s. These parameters are more independent of interproximal wear and loss of lateral enamel. PMID- 21624639 TI - Midwives count: achieving effective use of a scarce resource. PMID- 21624640 TI - Evaluation of selenium in biological sample of arsenic exposed female skin lesions and skin cancer patients with related to non-exposed skin cancer patients. AB - The antagonistic effects between selenium (Se) and arsenic (As) suggest that low Se status plays an important role in arsenism development. The objective of present study was to assess Se contents in biological samples of As exposed females have skin lesions and cancer with related to non-exposed skin cancer patients. The biological samples (blood and scalp hair) of As exposed group comprises, female skin cancer (ESC) patients admitted in cancer hospitals have skin lesions (ESL) and exposed referents have not both diseases (ER), belongs to As exposed area of Pakistan. For comparative purposes, age matched female skin cancerous patient (RP) and non-cancerous females (NER) belong to non-exposed areas were also selected. The As and Se in acid digests of biological samples were pre-concentrated by complexing with chelating agent (ammonium pyrrolidinedithiocarbamate), and resulted complexes were extracted into non-ionic extractant (Triton X-114), prior to analysis by electrothermal atomic absorption spectrometry. The enhancement factor of about 25 was obtained by pre concentrating 10 mL of sample solutions. The accuracy of the optimized procedure was evaluated by using certified reference material (BCR 397) with certified values for Se and As and standard addition method at three concentration levels in real samples. No significant differences was observed (p>0.05) when comparing the values obtained by the proposed method, added and certified values of both elements. The biological samples of ESC patients had 2-3 folds higher As and lower Se levels as compared to RP (p<0.001). Understudied exposed referents have high level of As and lower Se contents as compared to referents subjects of non exposed area (p<0.01). The higher concentration of As and lower levels of Se in biological samples of cancerous patients are consisted with reported studies. PMID- 21624641 TI - Infection of the Sunagoke moss panels with fungal pathogens hampers sustainable greening in urban environments. AB - Drought and heat tolerance of the Sunagoke moss (Racomitrium japonicum) and the low thermal conductivity of the dry moss tissue offer novel greening and insulation possibilities of roofs and walls to mitigate the heat island phenomenon in urban environments. However, damage may appear in the moss panels under humid conditions in Japan. In this study we characterized fungi associated with the damaged areas of the Sunagoke moss panels. Fungi were identified by morphology and internal transcribed spacer (ITS) sequence analysis and tested for pathogenicity on R. japonicum (Grimmiaceae) and an unrelated moss species (Physcomitrella patens; Funariaceae) under controlled conditions. Alternaria alternata, Fusarium avenaceum and Fusarium oxysporum caused severe necrosis and death, whereas Cladosporium oxysporum and Epicoccum nigrum caused milder discoloration or chlorosis in both moss species. The fungi pathogenic on moss were closely related to fungal pathogens described from cultivated vascular plants. Ammonium increased severity of fungal diseases in moss. This study demonstrated that fungi can cause economically significant diseases in cultivated moss and hamper commercial use of the moss panels unless appropriate control methods are developed. Use of a single moss clone to cover large surfaces and the air pollutants such as ammonium may increase the risk for fungal disease problems. PMID- 21624642 TI - Changes in water quality of the River Frome (UK) from 1965 to 2009: is phosphorus mitigation finally working? AB - The water quality of the River Frome, Dorset, southern England, was monitored at weekly intervals from 1965 until 2009. Determinands included phosphorus, nitrogen, silicon, potassium, calcium, sodium, magnesium, pH, alkalinity and temperature. Nitrate-N concentrations increased from an annual average of 2.4 mg l-1 in the mid to late 1960s to 6.0 mg l-1 in 2008-2009, but the rate of increase was beginning to slow. Annual soluble reactive phosphorus (SRP) concentrations increased from 101 MUg l-1 in the mid 1960s to a maximum of 190 MUg l-1 in 1989. In 2002, there was a step reduction in SRP concentration (average=88 MUg l-1 in 2002-2005), with further improvement in 2007-2009 (average=49 MUg l-1), due to the introduction of phosphorus stripping at sewage treatment works. Phosphorus and nitrate concentrations showed clear annual cycles, related to the timing of inputs from the catchment, and within-stream bioaccumulation and release. Annual depressions in silicon concentration each spring (due to diatom proliferation) reached a maximum between 1980 and 1991, (the period of maximum SRP concentration) indicating that algal biomass had increased within the river. The timing of these silicon depressions was closely related to temperature. Excess carbon dioxide partial pressures (EpCO2) of 60 times atmospheric CO2 were also observed through the winter periods from 1980 to 1992, when phosphorus concentration was greatest, indicating very high respiration rates due to microbial decomposition of this enhanced biomass. Declining phosphorus concentrations since 2002 reduced productivity and algal biomass in the summer, and EpCO2 through the winter, indicating that sewage treatment improvements had improved riverine ecology. Algal blooms were limited by phosphorus, rather than silicon concentration. The value of long-term water quality data sets is discussed. The data from this monitoring programme are made freely available to the wider science community through the CEH data portal (http://gateway.ceh.ac.uk/). PMID- 21624643 TI - Thrombin generation in chronic obstructive pulmonary disease: dependence on plasma factor composition. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is associated with an increased risk for thromboembolic events. We investigated thrombin generation profiles in COPD patients and their dependence on plasma factor/inhibitor composition. METHODS: Factors (f) (fII, fV, fVII, fVIII, fIX, fX), antithrombin, protein C (PC) and free tissue factor pathway inhibitor (fTFPI) from 60 COPD patients (aged 64.2 +/- 10.1 years; a mean forced expiratory volume in 1 second [FEV(1)], 55.6 +/- 15.8% of predicted values) were compared with those for 43 controls matched for age, sex, weight and smoking. Patients receiving anticoagulation were excluded. Using each individual's plasma coagulation protein composition, tissue factor-initiated thrombin generation was assessed computationally. RESULTS: COPD patients had higher fII (115 +/- 16 vs 102 +/- 10%, p < 0.0001), fV (114 +/- 19 vs 102 +/- 12%, p = 0.0002), fVII (111 +/- 15 vs 102 +/- 17%, p = 0.002), fVIII (170 +/- 34 vs 115 +/- 27%, p < 0.0001), and fIX (119 +/- 21 vs 107 +/- 17%, p = 0.003), and lower fTFPI (17.7 +/- 3.2 vs 18.9 +/- 3.2 ng/ml, p = 0.047) compared with controls, while fX, antithrombin, and PC were similar in both groups. Computational thrombin generation profiles showed that compared with controls, COPD patients had higher maximum thrombin levels (+28.3%, p < 0.0001), rates of thrombin generation (+46.1%, p < 0.0001) and total thrombin formation (+14.4%, p < 0.001), together with shorter initiation phase of thrombin generation (p < 0.0001) and the time to maximum thrombin levels (p < 0.0001). Thrombin generation profiles in COPD patients can be normalized via correction of fII, fVIII , fIX and TFPI. The severity of COPD and inflammatory markers were not associated with thrombin generation profiles. CONCLUSIONS: Prothrombotic phenotype in COPD patients is largely driven by increased prothrombin, fVIII, fIX, and lower fTFPI. PMID- 21624644 TI - Epoprostenol inhibits human platelet-leukocyte mixed conjugate and platelet microparticle formation in whole blood. AB - Circulating platelet-leukocyte mixed conjugates and platelet microparticles are potential markers of inflammation in the atherothrombotic disease. Epoprostenol is a synthetic salt of PGI2 (prostacyclin) clinically used in pulmonary hypertension and transplantation as a potent inhibitor of platelet aggregation. In this study the in vitro effect of this drug was investigated on the interaction of platelets with leukocytes and on markers of leukocyte and platelet activation, including platelet microparticle formation. The analyses were performed by flow cytometry on citrated whole blood collected from healthy subjects and challenged by a mixture of collagen-ADP. Preliminarily, the epoprostenol antiplatelet effect was confirmed by both aggregometry and PFA-100 and by evaluation of intraplatelet VASP phosphorylation. Epoprostenol, at nanomolar concentrations, prevented the formation of platelet mixed conjugates with PMN or monocytes, platelet PAC-1 and P-selectin expression and platelet microparticle generation. The reference drugs PGE1, aspirin and the novel ADP receptor antagonist, cangrelor, were only effective at micromolar concentrations. No effect of epoprostenol was detected on leukocyte activation markers. Our data suggest a possible additional mechanism of action of epoprostenol in reducing the inflammatory cell contribution to pulmonary hypertension and thrombosis. PMID- 21624645 TI - Local regulation of neutrophil elastase activity by endogenous alpha1-antitrypsin in lipopolysaccharide-primed hematological cells. AB - Neutrophil elastase released from activated neutrophils contributes in combating bacterial infection. While chronic inflammation results in anemia and decreased bone marrow activities, little is known about the effect of neutrophil elastase on hematological cell growth in severe inflammatory states. Here, we demonstrated that alpha1-antitrypsin, a physiological inhibitor of neutrophil elastase, functions as a regulator for cell growth by neutralizing neutrophil elastase activity in lipopolysaccharide-primed hematological cells. HL-60 cells were resistant to neutrophil elastase, as they also expressed alpha1-antitrypsin. The growth of HL-60 cells transduced with a LentiLox-short hairpin alpha1-antitrypsin vector was significantly suppressed by neutrophil elastase or lipopolysaccharide. When CD34(+) progenitor cells were differentiated towards a granulocytic lineage, they concomitantly expressed neutrophil elastase and alpha1-antitrypsin and prevented neutrophil elastase-induced growth inhibition. These results suggest that granulocytes might protect themselves from neutrophil elastase-induced cellular damage by efficiently neutralizing its activity through the simultaneous secretion of endogenous alpha1-antitrypsin. PMID- 21624646 TI - Intraoperative frozen section analysis of urethral margin biopsies during radical prostatectomy. AB - OBJECTIVES: To evaluate the accuracy and potential clinical value of intraoperative frozen section analysis (FSA) on urethral margin (UM) tissue during radical prostatectomy. Positive surgical margins increase the risk of post operative cancer recurrence. Positive surgical margins are frequently found at the apex. The utility of intraoperative FSA of the margins is controversial. METHODS: We reviewed a consecutive series of radical prostatectomy cases (n = 1669) performed at our institution, in which UMs were routinely evaluated by intraoperative FSA. RESULTS: The submitted UM tissue contained cancer glands in 111 cases (6.7%). On FSA, the pathologists detected cancer in 55 cases (3.3%), missed cancer in 38 (2.3%), and reported atypical glands in 18 (1.1%). FSA of the UMs had a sensitivity of 59.1%, specificity of 99.8%, and positive and negative predictive value of 94.8% and 97.6%, respectively. The low sensitivity resulted from a substantial false-negative rate (n = 38), which was largely attributed to limited sampling on FSA (n = 31). Of the 55 patients (3.3%) whose positive UMs were detected by FSA, 20 (1.2%) had cancer-free margins after tissue re-excision. A positive final UM was associated with greater biochemical recurrence (P = .0073). However, the few patients limited the statistical analysis of the benefit of margin conversion through tissue re-excision (P = .35). CONCLUSIONS: Although experienced pathologists can evaluate the UMs on FSA with good accuracy, FSA has a relatively low sensitivity. Our data have indicated a low yield and a questionable value of routine FSA during radical prostatectomy. PMID- 21624647 TI - Functional, metabolic, and morphologic characteristics of a novel rat model of type 2 diabetes-associated erectile dysfunction. AB - OBJECTIVES: To conduct a pilot study to investigate functional, metabolic, and penile morphologic changes in a novel model of lean DM2. Erectile dysfunction (ED) is a frequent sequela in patients with type 2 diabetes mellitus (DM2). METHODS: Eight rats received a high-fat diet and 2 weeks later, 2 intraperitoneal injections of streptozotocin (STZ, 30 mg/kg). Five age-matched rats served as controls. Insulin challenge tests were performed at 6 and 12 weeks after induction of DM2. At 12 weeks, erectile function was tested by measurement of intracavernous pressure (ICP) increase upon cavernous nerve stimulation. Penile tissue and serum samples were harvested for histology and biochemistry, respectively. RESULTS: A lean DM2 model was established as demonstrated by decreased insulin resistance, elevated nonfasting plasma glucose levels, hyperlipidemia, and decreased insulin concentration in the absence of obesity. ICP/mean arterial pressure was significantly decreased in DM2 animals (0.29) compared with controls (0.81). Expression of neuronal nitric oxide synthase and rat endothelial cell antigen-1, and the smooth muscle/collagen ratio were significantly decreased in the penis of DM2 animals. CONCLUSIONS: We propose an inexpensive nongenetic animal model of lean DM2-associated ED. Microanatomical changes in the erectile tissue that reflect an advanced stage of the disease were observed. PMID- 21624648 TI - Anaphylaxis to diphtheria, tetanus, and pertussis vaccines among children with cow's milk allergy. PMID- 21624649 TI - Efficacy and tolerability of systemic methylprednisolone in children and adolescents with chronic rhinosinusitis: a double-blind, placebo-controlled randomized trial. AB - BACKGROUND: The place of systemic corticosteroids in the treatment of children with chronic rhinosinusitis (CRS) remains unclear. OBJECTIVE: We sought to assess the effectiveness and tolerability of oral methylprednisolone as an anti inflammatory adjunct in the treatment of CRS in children. METHODS: Forty-eight children (age, 6-17 years) with clinically and radiologically proved CRS were included. Patients were randomly assigned to either oral amoxicillin/clavulanate (AMX/C) and methylprednisolone or AMX/C and placebo twice daily for 30 days. Oral methylprednisolone was administered for the first 15 days with a tapering schedule. Primary parameters were mean change in symptom and sinus computed tomographic (CT) scan scores after treatment. Secondary study parameters were mean changes in individual symptom scores after treatment, relapse rate, and tolerability. RESULTS: Forty-five patients completed the study: 22 received AMX/C and methylprednisolone, and 23 received AMX/C and placebo. Both groups demonstrated significant improvements in symptom and sinus CT scores when comparing baseline values with end-of-treatment values (P < .001). Methylprednisolone as an adjunct was significantly more effective than placebo in reducing CT scores (P = .004), total rhinosinusitis symptoms (P = .001), and individual symptoms of nasal obstruction (P = .001), postnasal discharge (P = .007), and cough (P = .009). At the end of treatment, 48% of the children in the placebo group still had abnormal findings on CT scans versus 14% in the methylprednisolone group (P = .013). Therapy-related adverse events were not different between groups. Although insignificant, the incidence of clinical relapses was also less in the methylprednisolone group (25%) compared with that in the placebo group (43%, P = .137). CONCLUSION: Oral methylprednisolone is well tolerated and provides added benefit to treatment with antibiotics for children with CRS. PMID- 21624650 TI - Receptor-targeted liposome-peptide nanocomplexes for siRNA delivery. AB - RNA interference induced by double-stranded, small interfering RNA (siRNA) molecules has attracted great attention as a genetic therapeutic approach. Despite major advances in this field, new nanoparticle formulations are required for in vivo delivery of siRNA, particularly for tissue-specific delivery of siRNA reagents. We have developed and optimized LYR nanocomplex formulations for siRNA delivery that consist of a liposome (DOTMA/DOPE; L) and a targeting peptide (K16GACYGLPHKFCG; Y) which self-assemble on mixing at optimal ratios with siRNA (R). Biophysical measurements indicated that LYR nanocomplexes were strongly cationic, mainly spherical particles of less than 100 nm. These formulations packaged and protected siRNA on incubation with RNAseA with >90% intact siRNA recovery. In addition, intact siRNA was recovered from LYRs upon heparin treatment. A critical synergy was observed between the lipid and peptide components for LYR particle stability and transfection efficiency. To evaluate targeting, transfections were compared with non-targeted formulations containing K16 with no targeting ligand. Gene knockdown efficiencies with targeted formulations were more than two-fold better in all cell lines tested (p < 0.01). LYR formulations with liposomes containing DOTMA, which has an 18-carbon (C18) alkyl tail, were significantly better in silencing than formulations containing cationic lipids with shorter alkyl tails. LYRs with siRNA against endogenous luciferase and GAPDH were successful in silencing these genes in 3 cell lines (1HAEo- human airway epithelial, B104 rat neuroblastoma, Neuro2A-Luc mouse neuroblastoma) in vitro with 80% efficiency, similar in efficiency to Lipofectamine 2000. Confocal microscopy analysis with LYRs containing fluorescently labelled siRNA (Cy3) showed that the siRNA was located in the perinuclear region of the cytoplasm, where the RNA-induced silencing complex (RISC) is likely to be found. The LYR formulations may have applications for the further development of siRNA-based therapeutics. PMID- 21624651 TI - Targeted delivery of non-viral vectors to cartilage in vivo using a chondrocyte homing peptide identified by phage display. AB - Gene therapy is a promising method for osteoarthritis and cartilage injury. However, specifically delivering target genes into chondrocytes is a great challenge because of their non-vascularity and the dense extracellular matrix of cartilage. In our study, we identified a chondrocyte-affinity peptide (CAP, DWRVIIPPRPSA) by phage display technology. Subsequent analysis suggests that the peptide can efficiently interact specifically with chondrocytes without any species specificity. Polyethylenimine (PEI) was covalently modified with CAP to construct a non-viral vector for cartilage-targeted therapy. To investigate the cartilage-targeting property of the CAP-modified vector, FITC-labeled CAP conjugated PEI/DNA particles were injected into rabbit knee joints, and visualized under confocal microscope. Higher concentrations of CAP-modified vector were detected in the cartilage and specifically taken up by chondrocytes compared with a randomly scrambled peptide (SP)-modified vector. To evaluate cartilage-targeting transfection efficiency, the GFP and luciferase genes were delivered into knee joints using CAP- and SP-modified PEI. Cartilage transfections mediated by CAP-modified PEI were much more efficient and specific than those by SP-modified PEI. This result suggests that CAP-modified PEI could be used as a specific cartilage-targeting vector for cartilage disorders. PMID- 21624652 TI - [Rediscovering the macula of high myopia in the XXI century]. PMID- 21624653 TI - [Impact of Nd: YAG capsulotomy on quality of life in pseudophakic patients]. AB - OBJECTIVE: To determine whether visual acuity (VA) improvement in patients with posterior capsule opacification (PCO), accepted after the capsulotomy, is followed by positive changes in the perception of their quality of life. METHOD: A total of 130 patients with PCO were examined before and after capsulotomy. In addition to the ophthalmic examination, the EuroQol questionnaire was completed. We analyzed the influence of demographic characteristics on the outcome of the test. RESULTS: The mean binocular visual acuity (BVA) and the mean scores in the EuroQol dimensions and EQ-VAS improved significantly after capsulotomy. VA did not have the same effect on daily common tasks. After treatment, the problems in the usual activities dimension have seen the largest reduction. Mobility is still the dimension with more problems regarding patients after capsulotomy. CONCLUSIONS: Decreased VA induced by PCO is associated with difficulties performing daily living activities. Capsulotomy results in a gain in VA and functional ability of the majority of patients, improving the perception of their quality of life. PMID- 21624654 TI - [Experimental study of pig dislocated intraocular lens shift after Nd: YAG laser capsulotomy]. AB - OBJECTIVES: To evaluate the movement from their initial set position of subluxated intraocular lenses (IOL) in pig eyes following Nd: YAG laser capsulotomy. METHODS: An extracapsular surgical intervention of the transparent crystalline lens was performed on 15 pig eyes. After the introduction of an intraocular lens into the capsular sac, a zonular dialysis was carried out to observe the shift of the IOL. We carried out a high intensity Nd: YAG laser capsulotomy in a 3.5mm cross shape in the central optical axis and then measured the final displacement of the IOL using a millimetre ruler. RESULTS: We did not observe a significant shift of the lens in any of the eyes studied. CONCLUSIONS: Nd: YAG laser capsulotomy is a safe short-term operation for static and subluxated intraocular lenses. PMID- 21624655 TI - [Comparative study of the pressure lowering efficacy and variations in the ocular pulse amplitude between fixed combinations of dorzolamide/timolol and brinzolamide/timolol]. AB - OBJECTIVE: To determine possible differences in the intraocular pressure (IOP) and ocular pulse amplitude (OPA) lowering capacity of the fixed drug combinations dorzolamide/timolol and brinzolamide/timolol. METHODS: In this cross-sectional study, one of the eyes of 25 healthy subjects was randomly assigned to treatment with dorzolamide/timolol and the other eye with brinzolamide/timolol. After instilling the drops, possible adverse effects (e.g., blurred vision, itching) were assessed in each eye. This assessment was repeated 30 minutes later. IOP and OPA were determined In each eye by dynamic contour tonometry at baseline and two hours following treatment. RESULTS: Both fixed drug combinations significantly reduced IOP and OPA with no differences detected between treatment groups. Among the adverse effects recorded, itching was significantly greater in the first assessment in the eyes treated with dorzolamide/timolol (P = .011). This difference was no longer apparent in the second assessment. CONCLUSIONS: Both fixed combinations were similarly effective in reducing intraocular pressure and ocular pulse amplitude. Adverse effects related to both treatments were mild and well-tolerated, though itching occurred most frequently in the eyes treated with dorzolamide/timolol. PMID- 21624656 TI - [Interferon alpha 2beta, partial keratectomy and amniotic membrane transplant for the treatment of a recurrent conjunctival squamous carcinoma]. AB - CASE REPORT: An 80-year-old woman diagnosed with a recurrent squamous conjuctival carcinoma treated with surgical excision of the lesion, zonal reconstruction and topical Interferon alpha 2-beta. DISCUSSION: Squamous conjunctival carcinoma is the most frequent neoplasm of the ocular surface. Surgical removal of the lesion is the traditional treatment, but this technique has a high recurrence rate. Interferons are glycoproteins that trigger intracellular pathways with antiviral and antitumoral properties. Recent studies have proven their activity against conjunctival carcinoma. PMID- 21624657 TI - [Giant optic disk melanocytoma complicated with massive intraocular seeding]. AB - CASE REPORT: A 33 year-old female with a large, unilateral, dark tumour of the optic disk with a presumed diagnosis of melanocytoma. After a follow-up of nineteen years, the tumour produced massive intraocular seeding with cataract, secondary glaucoma (pigmentary, melanocytomalytic, inflammatory and pupillary seclusion glaucoma) and amaurosis. The eye is enucleated and the pathological examination confirmed the diagnosis. DISCUSSION: Optic disk melanocytoma is a benign stationary tumour. However, sometimes that are to complications, including visual loss. Our case confirms that vitreous seeding is due to tumour necrosis with dissemination of tumoral debris and melanin, which are phagocytized by macrophages and produce an inflammatory reaction, trabecular plugging and ocular hypertension. PMID- 21624658 TI - [Rituximab for treatment of graves orbitopathy]. PMID- 21624659 TI - [Evaluation of the improvement in sensitivity with the new IOL-Master((r))]. PMID- 21624660 TI - [The enigmatic eye of Nefertiti]. PMID- 21624661 TI - Corticotropin-releasing factor binding protein enters the regulated secretory pathway in neuroendocrine cells and cortical neurons. AB - Corticotropin releasing factor binding protein (CRF-BP) is a 37kDa glycoprotein that binds CRF with high affinity. CRF-BP controls CRF levels within plasma during human pregnancy. It has also been shown that CRF-BP is expressed in various brain nuclei. Main actions that have been proposed for brain CRF-BP are either decreasing available CRF or facilitating CRF ligand-induced activation of CRF-R2 receptors. For both actions, it is necessary the release of CRF-BP from CRF-BP expressing neurons. However, the secretion mode of CRF-BP is currently unknown. We used heterologous expression of CRF-BP-Flag in PC12 cells and in primary culture of rat cortical neurons to study CRF-BP secretion mode. We observed that CRF-BP-Flag immunoreactivity presents the typical cytoplasmatic punctuate pattern that has been described for neuropeptides and proteins that enter the regulated secretory pathway in PC12 cells. Quantitative analysis of double immunofluorescence confocal images showed that CRF-BP-Flag colocalizes with secretogranin II, marker of secretory granules, both in PC12 and in primary cultured rat neurons. Furthermore, CRF-BP-Flag is released from PC12 cells upon high K(+)-depolarization. Thus, our results show that CRF-BP is efficiently sorted to the regulated secretory pathway in two cellular contexts, suggesting that the extracellular levels of CRF-BP in the central nervous system depends on neuronal activity. PMID- 21624662 TI - Molecular profiling--a tool for addressing emerging gaps in the comparative risk assessment of GMOs. AB - Assessing the risks of genetically modified organisms (GMOs) is required by both international agreement and domestic legislation. Many view the use of the "omics" tools for profiling classes of molecules as useful in risk assessment, but no consensus has formed on the need or value of these techniques for assessing the risks of all GMOs. In this and many other cases, experts support case-by-case use of molecular profiling techniques for risk assessment. We review the latest research on the applicability and usefulness of molecular profiling techniques for GMO risk assessment. As more and more kinds of GMOs and traits are developed, broader use of molecular profiling in a risk assessment may be required to supplement the comparative approach to risk assessment. The literature-based discussions on the use of profiling appear to have settled on two findings: 1. profiling techniques are reliable and relevant, at least no less so than other techniques used in risk assessment; and 2. although not required routinely, regulators should be aware of when they are needed. The dismissal of routine molecular profiling may be confusing to regulators who then lack guidance on when molecular profiling might be worthwhile. Molecular profiling is an important way to increase confidence in risk assessments if the profiles are properly designed to address relevant risks and are applied at the correct stage of the assessment. PMID- 21624663 TI - Complement deficiency states and associated infections. AB - A major function of the immune system is to protect the host from microbial infections. The complement system plays important roles in both the innate and the adaptive immune defense and also acts as a bridge between these arms of immunity. This is obvious from complement deficiencies which in varying degree, depending on which factor is missing, are associated with increased infection susceptibility and also increased risk for other, mainly autoimmune diseases. Genetically determined deficiencies are described for almost all complement proteins but the consequences show a wide variation. Here the genetic defects and molecular abnormalities in complement deficient persons, related clinically relevant infections and the options for prevention and therapy are reviewed. The roles of complement in host defense against common infections are also discussed. PMID- 21624664 TI - Reversion rates of QuantiFERON-TB Gold are related to pre-treatment IFN-gamma levels. AB - OBJECTIVE: The quantitative interferon (IFN)-gamma in response to Mycobacterium tuberculosis-specific antigens declines in tuberculosis patients after starting treatment, however, in some cases remains high despite clinical improvements. Our aim was to evaluate clinical parameters associated with remaining QuantiFERON-TB Gold (QFT-G) positive after treatment. METHODS: A prospective cohort study of 101 culture-positive, positive QFT-G, HIV-uninfected patients with pulmonary tuberculosis. QFT-G was performed at diagnosis, at the end of intensive phase, at treatment completion, and 5-7 months post-treatment completion. RESULTS: There were 80 patients with complete results, 34 (43%) remaining QFT-G positive and 46 (58%) reverting to QFT-G negative at the 5-7 month post-treatment time point. There was a significant decline in IFN-gamma levels in response to both CFP-10 and ESAT-6 with tuberculosis treatment. Multivariate analysis revealed significant associations between IFN-gamma levels detected before treatment and remaining QFT-G positive post-treatment after adjustment for smear status, presence of cavitation, and positive sputum culture two months after starting treatment. CONCLUSIONS: Quantitative QFT-G responses drop significantly in active tuberculosis patients undergoing treatment, with almost 60% becoming test negative. Reversion to a negative QFT-G result was closely associated with the magnitude of the IFN-gamma response prior to treatment and increasing age. PMID- 21624665 TI - Let us go to the video: part II. PMID- 21624666 TI - Comments on "comparison of 2 surgical techniques for reconstructing posterolateral corner of the knee: a cadaveric study evaluated by navigation system". PMID- 21624668 TI - Accuracy of intra-articular glenohumeral injections: the anterosuperior technique with arthroscopic documentation. AB - PURPOSE: Our objective was to assess the accuracy rate of needle placement with the anterosuperior technique of glenohumeral joint injection that uses familiar palpable superficial landmarks as a guide instead of diagnostic imaging. METHODS: Between April 2007 and October 2007 at our institution, 42 patients met the study inclusion criteria of being aged 18 years or older and undergoing shoulder arthroscopy. For the injection (performed by 1 surgeon), anesthetized patients were placed in the beach-chair position with the arm in adduction and internal rotation. The surgeon was allowed to redirect the needle only once without withdrawing the needle from the entry site. After injection, arthroscopic confirmation of needle position in the joint and the presence of backflow from the posterior portal cannula were used to determine accuracy and the relation of the needle to adjacent anatomy. RESULTS: Of the 42 injections, 38 needles were inserted accurately into the glenohumeral joint (91% accuracy rate), most through the rotator interval (21) or the long head of the biceps tendon (9). Four needles were placed inaccurately into the anterior synovium and subacromial space. Adhesive capsulitis was the diagnosis in 3 of those 4 shoulders but in only 5 of the 38 shoulders in the group with accurate placement (P < .05). Body mass index was not statistically different between the accurate and inaccurate injection groups (P > .05). CONCLUSIONS: Anterosuperior glenohumeral joint injection without image guidance provides an accuracy rate of 91%. The anterosuperior technique for glenohumeral injections yields an accuracy rate higher than that of the standard anterior techniques and comparable to that of posterior injection. LEVEL OF EVIDENCE: Level IV, diagnostic study. PMID- 21624669 TI - The effect on loop elongation and stress relaxation during longitudinal loading of FiberWire in shoulder arthroscopic knots. AB - PURPOSE: This study examined the viscoelastic properties of 6 common arthroscopic sliding knots (Tennessee slider, Roeder knot, SMC knot, Duncan loop, Weston knot, and Nicky's knot) with 3 reversing half-hitches on alternating posts, tied with No. 2 FiberWire (Arthrex, Naples, FL). Knot configuration was designed to simulate a double-row rotator cuff repair with suture bridges. METHODS: Constructs were loaded in 20-N increments to 100 N and held for 2 minutes to monitor the viscoelastic behavior in tension. Suture was also tested without tying a knot. RESULTS: Stress relaxation increased with loading but did not differ between knot configurations. Initial elongation was highest during the first loading to 20 N. Relaxation was greater for the Roeder knot at 20 N and for the Roeder and SMC knots at 80 N (P < .05) when compared with the loop with no knot. Elongation was greatest for the Roeder knot throughout all loads. This difference was significant at 60 N compared with the knotless loop. At 100 N, all knots showed greater elongation than the knotless loop (P < .05). Testing of suture, without any knots, accounted for more than 75% of the overall stress relaxation and loop elongation of the suture-knot construct. CONCLUSIONS: In our in vitro evaluation of the Tennessee slider, Roeder knot, SMC knot, Duncan loop, Weston knot, and Nicky's knot in a simulated suture bridge construct, knot configuration was not a variable that influenced elongation or stress relaxation. Overall response was primarily due to the suture itself. With the exception of the Roeder knot, relaxation was similar provided that a secure knot was formed at the time of original tying. CLINICAL RELEVANCE: With the evolution of surgical devices, the reliance on knots is decreasing. The results of this study suggest that using knotless techniques for securing the rotator cuff will not change the stress relaxation characteristics of the suture bridge. PMID- 21624670 TI - Do iatrogenic punctures of the labrum affect the clinical results of hip arthroscopy? AB - PURPOSE: To document the effect of iatrogenic labral punctures (ILPs) on the clinical results of hip arthroscopy. METHODS: From a database of 250 consecutive patients who had their hip arthroscopy performed by the senior author, 50 patients with ILPs were identified. Results of hip arthroscopy in these patients were compared with those of a matched group of 50 patients who did not have labral puncture (NLP). All hips were assessed with Byrd's 100-point modified Harris hip scoring system before arthroscopy and at 3, 6, 12, and 24 months after surgery. RESULTS: The mean age of the ILP and NLP patients was 40 and 36 years, respectively, and their preoperative scores averaged 36 points. Mean joint distraction was 13 mm and 15 mm in the ILP and NLP patients, respectively, and a positive "Byrd's sign" was seen in both the ILP (84%) and NLP (42%) patients. After surgery, the ILP and NLP patients had 6-month scores that averaged 85 and 88 points, respectively. At 12 months, their scores averaged 88 and 90 points, respectively, and after 24 months, their scores averaged 88 and 89 points, respectively. At all follow-up intervals, there were no significant differences between the scores of the 2 groups (P > .05). CONCLUSIONS: ILPs did not affect the 1- and 2-year clinical results of patients who sustained these injuries during hip arthroscopy. LEVEL OF EVIDENCE: Level III, case-control study. PMID- 21624671 TI - Treatment of athletes with symptomatic intra-articular hip pathology and athletic pubalgia/sports hernia: a case series. AB - PURPOSE: The purpose of the study was to evaluate the results of surgical treatment in athletes with associated intra-articular hip pathology and extra articular sports pubalgia. METHODS: Between December 2003 and September 2009, 37 hips (mean patient age, 25 years) were diagnosed with both symptomatic athletic pubalgia and symptomatic intra-articular hip joint pathology. There were 8 professional athletes, 15 collegiate athletes, 5 elite high school athletes, and 9 competitive club athletes. Outcomes included an evaluation regarding return to sports and modified Harris Hip Score, Short Form 12 score, and visual analog scale score. RESULTS: We evaluated 37 hips at a mean of 29 months (range, 12 to 78 months) after the index surgery. Thirty-one hips underwent thirty-five athletic pubalgia surgeries. Hip arthroscopy was performed in 32 hips (30 cases of femoroacetabular impingement treatment, 1 traumatic labral tear, and 1 borderline dysplasia). Of 16 hips that had athletic pubalgia surgery as the index procedure, 4 (25%) returned to sports without limitations, and 11 (69%) subsequently had hip arthroscopy at a mean of 20 months after pubalgia surgery. Of 8 hips managed initially with hip arthroscopy alone, 4 (50%) returned to sports without limitations, and 3 (43%) had subsequent pubalgia surgery at a mean of 6 months after hip arthroscopy. Thirteen hips had athletic pubalgia surgery and hip arthroscopy at one setting. Concurrent or eventual surgical treatment of both disorders led to improved postoperative outcomes scores (P < .05) and an unrestricted return to sporting activity in 89% of hips (24 of 27). CONCLUSIONS: When surgery only addressed either the athletic pubalgia or intra-articular hip pathology in this patient population, outcomes were suboptimal. Surgical management of both disorders concurrently or in a staged manner led to improved postoperative outcomes scoring and an unrestricted return to sporting activity in 89% of hips. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 21624672 TI - Tunnel widening after anterior cruciate ligament reconstruction: a prospective randomized computed tomography--based study comparing 2 different femoral fixation methods for hamstring graft. AB - PURPOSE: We prospectively sought to compare the incidence and properties of tunnel widening in patients undergoing anterior cruciate ligament reconstruction with quadrupled hamstring graft by use of either EndoButton CL (Smith & Nephew Endoscopy, Andover, MA) or Transfix (Arthrex, Naples, FL) on the femoral side with a bioabsorbable interference screw in the tibial tunnel by computed tomography scan. METHODS: We included 34 patients in the study and randomized them into 2 groups--EndoButton and Transfix groups. An anteromedial portal technique was used to create the femoral tunnels in the EndoButton group, whereas a transtibial technique was used in the Transfix group. A bioabsorbable screw was used on the tibial side in both groups. Thirty patients completed the study protocol, and assessment was done at regular intervals until 12 months of follow up was reached. The patients underwent computed tomography scans at 2 weeks, 3 months, and 6 months postoperatively. The diameters of the tunnels were measured perpendicular to the long axis of the tunnels on oblique coronal and oblique sagittal planes at 3 levels: aperture, midway, and suspension point. We performed functional scoring with the International Knee Documentation Committee 2000 Subjective Knee Evaluation score and Lysholm score. RESULTS: Femoral tunnel widening at the aperture and at midway was significantly greater in the EndoButton group compared with the Transfix group. A decrease in the loop length in the EndoButton group was associated with lesser tunnel widening, although this was not found to be statistically significant with the numbers available. A trend toward decreased tunnel widening at the aperture on the tibial side was observed when the tip of the screw was 10 to 15 mm away from the aperture. CONCLUSIONS: Femoral tunnel widening was significantly less in the Transfix group compared with the EndoButton group. LEVEL OF EVIDENCE: Level II, prospective comparative study. PMID- 21624673 TI - Outcomes after arthroscopic excision of the bony prominence in the treatment of tibial spine avulsion fractures. AB - PURPOSE: The purpose of this study was to determine the outcomes after arthroscopic excision of the bony prominence after a tibial spine avulsion fracture. METHODS: This study included 7 subjects (5 female and 2 male subjects; mean age, 21.4 years). All subjects underwent preoperative rehabilitation focused on range of motion (ROM) and swelling control. Postoperative rehabilitation focused on regaining symmetric knee hyperextension and flexion. Objective examinations and subjective surveys were obtained at least 1 year after surgery. RESULTS: All subjects achieved normal knee extension; 6 patients achieved normal knee flexion, whereas 1 patient had nearly normal flexion. Physical examination showed a negative Lachman test with a firm end point in all patients, and the mean side-to-side difference for the KT-1000 manual maximum test (MEDmetric, San Diego, CA) was 1.3 mm. No subjects required subsequent anterior cruciate ligament reconstruction. All subjects returned to their previous level of activity without instability symptoms. At a mean of 5.7 years after surgery, the mean International Knee Documentation Committee subjective survey score was 90.6 points overall, with 4.7 out of 5 possible points for the instability question. At latest follow-up, the mean ROM was from 6 degrees of hyperextension to 147 degrees of flexion in the involved knee, compared with 6 degrees of hyperextension to 148 degrees of flexion for the noninvolved knee. CONCLUSIONS: The results of arthroscopic excision of the bony fragment after type II, III, or III+ tibial spine avulsion fracture are positive, with good stability, symmetric ROM, and high subjective scores. Most importantly, this procedure allows patients to regain full, symmetric hyperextension of the knee, avoiding the complications associated with extension loss. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 21624674 TI - What effect does anterior cruciate ligament tibial guide orientation have on tibial tunnel length? AB - PURPOSE: To evaluate the effects of alteration in tibial guide pin insertion angle and external starting point on tibial tunnel length for anterior cruciate ligament (ACL) reconstruction. METHODS: Ten cadaveric tibial specimens were used. One pin was placed at each of variable insertion angles (55 degrees , 50 degrees , and 45 degrees ) of the tibial targeting device aimed at the center of the tibial ACL footprint. These 3 pins started externally along the anterior border of the superficial medial collateral ligament. A fourth pin at 50 degrees was placed at a different external tibial starting point 1.5 cm anterior to the anterior border of the superficial medial collateral ligament. The intraosseous length of each pin was measured. Statistic analyses were performed with the Kruskal-Wallis test, with significance set at P < .05. RESULTS: The mean length for the 55 degrees tibial tunnel was 50.3 mm (range, 42 to 56 mm); for the 50 degrees tunnel, it was 48.9 mm (range, 44 to 55 mm); for the 50 degrees anterior tunnel, it was 47.6 mm (range, 39 to 55 mm); and for the 45 degrees tunnel, it was 47.3 mm (range, 41 to 52 mm). Changing the angle of the tibial guide did not significantly affect the length of the tibial tunnel (P = .18). Changing the external tibial starting point did not affect the length of the tibial tunnel (P = .39). CONCLUSIONS: Changing the tibial guide angle between 45 degrees , 50 degrees , and 55 degrees does not appreciably change tibial tunnel length. Moving the starting point anterior 1.5 cm toward the tibial tubercle also has no effect on the tibial tunnel length. The lack of significant changes in tunnel length with these interventions may reflect the associated changes that occur in proximal tibial morphometry with change in external tibial starting position. CLINICAL RELEVANCE: Changing tibial tunnel length in ACL reconstruction likely requires more distalization of the external tibial starting point than is achieved simply by altering the tibial aiming guide angle by 10 degrees or less. PMID- 21624675 TI - Arthroscopic fixation of matrix-associated autologous chondrocyte implantation: importance of fixation pin angle on joint compression forces. AB - PURPOSE: The aim of the study was to investigate the effect of pin fixation perpendicular and 30 degrees tilted to the matrix surface on the joint compression forces. METHODS: In a porcine knee model, joint compression forces were recorded with a digital pressure sensor above the medial meniscus and with axial compression of 100 N by use of a material testing machine. The forces were recorded for an intact femoral condyle, as well as a standardized cartilage defect of 25 * 20 mm, after matrix-associated autologous chondrocyte implantation (m-ACI) (BioSeed C; Biotissue Technologies, Freiburg, Germany), fixed by use of a conventional suture technique and pin fixation with a biodegradable pin perpendicular and 30 degrees tilted to the matrix surface. RESULTS: In knees with cartilage defects, the peak compression forces (mean, 824 kPa) were significantly increased compared with the intact knee joint (564 kPa). After m ACI implantation with a chondral suture (581.3 kPa) and perpendicular pin fixation, the joint compression forces of the cartilage defect were significantly decreased (630.7 kPa). There were no significant differences compared with the intact knee. After 30 degrees tilted pin insertion, mean joint compression forces were significantly increased (1,740 kPa). CONCLUSIONS: This study shows that after chondral suture and perpendicular pin fixation, there are no increased compression forces in the knee joint in comparison to an intact knee. Thirty degree tilted pin insertion contributes to increased joint compression forces. CLINICAL RELEVANCE: A tilted insertion during pin fixation in m-ACI should be avoided because it may lead to increased joint compression forces, especially after cartilage defect lesions on the tibial side. PMID- 21624676 TI - Effectiveness of a footprint guide to establish an anatomic femoral tunnel in anterior cruciate ligament reconstruction: computed tomography evaluation in a cadaveric model. AB - PURPOSE: To compare drilling the femoral tunnel with an offset aimer and BullsEye guide (ConMed Linvatec, Largo, FL) to perform an anatomic single-bundle reconstruction of the anterior cruciate ligament (ACL) through the anteromedial portal. METHODS: Seven matched pairs of cadaveric knees were studied. The intent was to drill the femoral tunnel anatomically in all cases. In group A the femoral tunnel was drilled arthroscopically with an offset aimer. In group B the femoral tunnel was drilled arthroscopically with the BullsEye guide. Two tunnels were drilled through the same entry point in each knee. One was done at 110 degrees of knee flexion and the other at 130 degrees . They were scanned by computed tomography and reconstructed 3-dimensionally. Volume-rendering software was used to document relations of the drilled tunnel to the bony anatomy and tunnel length. RESULTS: In group B the femoral tunnel was placed at the center of the femoral insertion site. The center of the tunnel was 9.4 mm from the high cartilage margin and 8.6 mm from the low cartilage margin. In group A the tunnels were placed deeper (5.4 mm and 12.6 mm, respectively) (P = .018). There were no differences in tunnel length for either knee flexion degree. Three of the tunnels drilled at 110 degrees in group A compromised the posterior tunnel wall and measured less than 25 mm in length. CONCLUSIONS: Accurate placement in the center of the femoral footprint of the ACL is better accomplished with the BullsEye guide rather than 5-mm offset aimers. Five-millimeter offset aimers might cause posterior tunnel blowout and present the risk of obtaining short tunnels when performing oblique femoral tunnel placement through the anteromedial portal at 110 degrees of knee flexion. CLINICAL RELEVANCE: The BullsEye guide might be better than standard offset aimers in the performance of anatomic single-bundle ACL reconstruction. PMID- 21624677 TI - Effect of varying the length of soft-tissue grafts in the tibial tunnel in a canine anterior cruciate ligament reconstruction model. AB - PURPOSE: To investigate the effect of graft length within the bone tunnel on tendon-bone healing at an early stage after anterior cruciate ligament (ACL) reconstruction using Achilles tendon autograft in a canine model. METHODS: We divided 40 adult dogs into 4 groups (n = 10 per group). Each dog underwent ACL reconstruction with Achilles tendon autograft in both knees. In groups I, II, III, and IV, the graft length within the tibia tunnel was 5 mm, 10 mm, 15 mm, and 20 mm, respectively. Five dogs in each group were killed 6 and 12 weeks postoperatively, with 3 knees used for histologic observation and 7 knees for mechanical testing. RESULTS: Six weeks after surgery, the histologic scores in group I were lower than those in the other groups (P < .01) and those in group II were lower than those in group III and group IV (P < .01). However, there was no difference between group III and group IV (P > .05). At 12 weeks, there were no significant differences in histologic scores between groups (P > .05). The biomechanical test at 6 weeks showed that the mean graft pullout strength improved from group I to group IV. Except between groups III and IV (P = .142), there was a significant difference in the pullout strength among other groups. At week 12, the failure points were in the midsubstance of most specimens and more tibial-side graft pullout was found in group I than in the other groups (P < .001). CONCLUSIONS: The histologic maturity and biomechanical strength of the tendon-bone junction after ACL reconstruction in dogs will be delayed at an early stage if the graft length in the bone tunnel is less than 15 mm. CLINICAL RELEVANCE: There is a minimal acceptable amount of intratunnel tendon graft to allow satisfactory early tendon-bone healing, and attempts to save tendon graft length that lower the amount of intratunnel graft below a minimum of 15 mm should be avoided. PMID- 21624678 TI - Bacterial adherence to high--tensile strength sutures. AB - PURPOSE: The purpose of this study was to evaluate the bacterial adherence to high--tensile strength suture materials using a bioluminescent in vitro model. METHODS: Eleven strands each of No. 2 MaxBraid (Arthrotek [Biomet], Warsaw, IN), FiberWire (Arthrex, Naples, FL), Ethibond (Ethicon, Somerville, NJ), Orthocord (DePuy Mitek, Raynham, MA), and silk (Ethicon) sutures were immersed in a broth of bioluminescent Staphylococcus aureus, which is genetically engineered to emit photons. After 12 hours in the broth, the suture strands were individually irrigated with 10 mL of low-pressure normal saline solution and imaged with a photon-capturing camera system that yields a total photon count that correlates directly with residual bacterial counts. RESULTS: MaxBraid had the greatest adherence, followed by FiberWire, Ethibond, Orthocord, and silk. Orthocord had only 25% of the bacterial adherence of MaxBraid (P < .001). Ethibond and FiberWire had 53% (P < .001) and 75% (P = .003) of the adherence of MaxBraid, respectively. Differences between each suture were also statistically significant, with Ethibond and Orthocord having 71% (P = .007) and 33% (P < .001) of the adherence of FiberWire, respectively, and Orthocord having 47% (P < .001) of the adherence of Ethibond. The adherence to silk was statistically lower than all of the high-tensile strength sutures. CONCLUSIONS: Among high--tensile strength sutures, Orthocord has significantly less bacterial adherence than MaxBraid and FiberWire. Although infections in arthroscopic shoulder surgery are rare, the physical properties of surgical implants should be known by surgeons. In addition, bacterial adherence may contribute to suture selection in a patient prone to infection or to the use of suture in other body areas at greater risk for contamination. CLINICAL RELEVANCE: Bacterial adherence to high--tensile strength sutures may be a useful factor in implant selection in a patient with predisposition for contamination or infection. PMID- 21624679 TI - Innovation: back to our roots. PMID- 21624680 TI - Prevention and management of stiffness after arthroscopic rotator cuff repair: systematic review and implications for rotator cuff healing. AB - PURPOSE: To define the incidence of stiffness after arthroscopic rotator cuff repair (ARCR) and address the prevention and management of postoperative stiffness. METHODS: A PubMed search was performed using the combined terms "rotator cuff and stiffness," "shoulder and capsular release," and "capsular release and stiffness." Inclusion criteria were (1) primary outcome of postoperative shoulder stiffness, (2) Levels I to IV evidence, and (3) entirely arthroscopic technique for rotator cuff repair. RESULTS: Seven articles met our inclusion criteria. Two articles discussed the management of preoperative stiffness and ARCR. These articles managed preoperative stiffness differently, preventing an adequate comparison to provide recommendations. Three articles described both the incidence of and risk factors for postoperative stiffness after ARCR. The incidence of transient stiffness responsive to nonoperative management was 10%. The incidence of resistant stiffness that was permanent or required capsular release was 3.3%. Several risk factors for stiffness were identified in this review. Two articles focused on postoperative rehabilitation protocols and stiffness after ARCR. Overall, resistant postoperative stiffness was reported in 1.5% of patients with an immediate passive range-of-motion protocol, 4.5% of patients in a 6-week sling-immobilization protocol, and 0% of patients with a modified protocol. Three articles, all among the above mentioned articles, reported that arthroscopic capsular release for stiffness after ARCR improved motion to a level comparable to that in patients who did not require reoperation. CONCLUSIONS: The studies in this systematic review indicate that with ARCR, postoperative stiffness resistant to nonoperative management is uncommon despite an initial immobilization period. In the setting of resistant postoperative stiffness, arthroscopic capsular release can successfully restore range of motion. ARCR allows a delayed mobilization protocol that may be important in achieving rotator cuff healing. The optimal management of preoperative stiffness in patients undergoing ARCR is inconclusive based on this review. LEVEL OF EVIDENCE: Level IV, systematic review of Level III and IV studies. PMID- 21624681 TI - In vitro and intraoperative laxities after single-bundle and double-bundle anterior cruciate ligament reconstructions. AB - PURPOSE: The purpose of this study was to objectively evaluate whether double bundle anterior cruciate ligament (ACL) reconstruction can better restore the normal translational and rotational laxities than the conventional single-bundle ACL reconstruction among the reported biomechanical studies. METHODS: A systematic literature search was conducted to identify in vitro and in vivo (intraoperative) biomechanical studies that compared the laxities (anterior or anteroposterior or rotational) between single- and double-bundle ACL reconstructions. Because of large variability among the loading conditions and testing methods used to determine the rotational laxities among the studies, a meta-analysis of rotational laxities was not feasible. RESULTS: Seven in vitro and three in vivo studies were included in this analysis based on the predefined inclusion criteria. The overall mean differences calculated by the random effects model in anteroposterior laxity between the single-bundle and double-bundle ACL reconstruction techniques at 0 degrees , 30 degrees , 60 degrees , and 90 degrees of flexion were 0.99 mm, 0.38 mm, 0.34 mm, and 0.07 mm, respectively. No statistically significant difference was noted between the 2 treatments at all flexion angles. Among the 9 studies that compared the rotational laxity of single bundle and double-bundle ACL reconstructions, 4 reported that double-bundle reconstruction can provide better rotational control than single-bundle reconstruction. The other 5 studies could not identify any significant difference between the 2 reconstructions in terms of rotational laxity. CONCLUSIONS: Both single- and double-bundle treatment options for ACL injury result in similar anteroposterior knee joint laxity at time 0. No conclusive evidence on the superiority of 1 reconstruction technique over the other in terms of rotation laxity can be obtained because of several variations in the experimental protocol and the kinematics used to measure the rotational laxity among the studies. LEVEL OF EVIDENCE: Level III, meta-analysis. PMID- 21624682 TI - Type A behavior pattern and hyperthymic temperament: possible association with bipolar IV disorder. AB - BACKGROUND: Type A behavior pattern (TABP) has traditionally been reported to be associated with coronary heart disease and, more recently, several researchers have examined its association with depression and bipolar disorder. According to Akiskal and Pinto (1999), there are 2 subtypes of bipolar spectrum which are not associated with manic or hypomanic state. These are bipolar II1/2 (depression in those who have cyclothymic temperament) and bipolar IV (depression in those who have hyperthymic temperament). Our hypothesis is that individuals with hyperthymic temperament may have a tendency towards TABP. OBJECTIVES: The purposes of the present study are to investigate the association between TABP and hyperthymic temperament and to determine other biological factors associated with TABP. METHODS: Fifty healthy subjects were assessed for TABP and hyperthymic temperament by self-rating scales, daily activity, sleep time and illuminance by actigraphy, and central serotonergic function via the neuroendocrine challenge test. Serum brain-derived neurotrophic factor (BDNF) levels were also measured. RESULTS: Stepwise regression analysis indicated that hyperthymic temperament score was positively associated with TABP scores and both sleep time and snooze time were negatively associated with TABP scores. BDNF levels were not associated with TABP scores. CONCLUSIONS: These findings suggest that individuals with hyperthymic temperament may have a tendency towards TABP, and TABP persons may have short sleep time and short snooze time. Although further studies are required to investigate the association between TABP and affective disorders, the present findings clearly indicate the association between TABP and hyperthymic temperament, which may be associated with bipolar IV disorder. Taking TABP as a risk factor of cardiovascular events into consideration, this association between TABP and bipolar IV disorder may account for the well-known cardiovascular mortality in bipolar disorder. PMID- 21624683 TI - Psychological resilience in OEF-OIF Veterans: application of a novel classification approach and examination of demographic and psychosocial correlates. AB - A growing number of studies have examined the prevalence and correlates of psychopathology in Veterans of Operations Enduring Freedom and Iraqi Freedom (OEF OIF), but few have examined determinants of resilience in this population. This study employed a novel approach to classify psychological resilience in a cross sectional sample of OEF-OIF Veterans. A total of 272 predominantly older reserve/National Guard OEF/OIF Veterans completed a mail survey that assessed combat exposure, psychopathology, psychosocial functioning, and aspects of social support. Cluster analysis of scores on measures of combat exposure and PTSD symptoms revealed that a three-group solution best fit the data: Controls (low combat exposure, low PTSD symptoms); PTSD (high combat exposure, high PTSD symptoms); and Resilient (high combat exposure, low PTSD symptoms). Compared to the PTSD group, the Resilient group was more likely to be in a relationship and active duty; they also scored lower on a measure of psychosocial dysfunction, and higher on measures of psychological resilience and postdeployment social support. Logistic regression analysis revealed that being in a relationship, having fewer psychosocial difficulties, and reporting greater perceptions of purpose/control and family support and understanding were significantly associated with resilient group membership. Results of this study demonstrate a novel approach to classifying psychological resilience and suggest that interventions to mitigate psychosocial difficulties, enhance perceptions of purpose and control, and bolster family support and understanding may help promote resilience to combat related PTSD in OEF-OIF Veterans. PMID- 21624684 TI - Evaluating the CANSAS self-report (CANSAS-P) as a screening instrument for care needs in people with psychotic and affective disorders. AB - We examined agreement between the CANSAS self-report version (CANSAS-P) and the Camberwell Assessment of Need (CAN) interview in 200 long-term patients with affective and psychotic disorders. Intra-class correlations were fair to good for unmet needs. Overall, more unmet needs were reported on the CANSAS-P than in the CAN interview. No differences were found for patients with psychotic versus affective disorders. We conclude from this that the CANSAS-P is a promising screening instrument to detect unmet needs in people with severe mental illnesses. PMID- 21624685 TI - Switching patients from warfarin to dabigatran therapy: to RE-LY or not to rely. PMID- 21624686 TI - Postnatal regression of hypertrophic cardiomyopathy in infants of diabetic mothers: a crosstalk between Hox genes and epidermal growth factor (EGF) gene polymorphism? PMID- 21624687 TI - Preface. PMID- 21624688 TI - The evolution of pediatric critical care nursing: past, present, and future. AB - Although current nursing literature is overflowing with information related to the history of nursing in general, and even pediatric nursing, very little is published about PICU nursing. The evolution of pediatric critical care nursing is presented based on a historical context, the current state, and future projections. More specifically, this treatise focuses on the environment, the patient and family, and of course, the PICU nurse. Concluding remarks provide an insight into how health care reforms and how the use of clinical information technology will affect the role of the pediatric critical care nurse in the future. PMID- 21624689 TI - Acute cardiac arrhythmias following surgery for congenital heart disease: mechanisms, diagnostic tools, and management. AB - This article focuses on the management of those cardiac arrhythmias most commonly seen in the immediate postoperative period. They include ventricular tachycardia, ventricular fibrillation, atrial flutter, junctional ectopic tachycardia, bradycardia, and atrioventricular block. The mechanisms of cardiac arrhythmias are reviewed followed by a brief overview of the predominant acute arrhythmias, tools used for the diagnostic evaluation of these arrhythmias, management strategies, and, finally, nursing considerations. PMID- 21624690 TI - Use of continuous glucose monitoring systems in children with type 1 diabetes. AB - Continuous glucose monitoring systems (CGMS) offer a recent technological solution to fear of hypoglycemia and reduction in hyperglycemia. With glucose readings every 5 minutes, and the alarm capabilities to signal rapid increases or decreases in blood glucose, they afford a sense of security for both children and parents. Although children and parents must continue to be vigilant in the treatment of diabetes, CGMS may effectively allow tight diabetes control with earlier detection of hypoglycemia onset. This article provides an overview of CGMS available at the time of publication; new generations of systems have been introduced annually with improvements in sensitivity and user friendliness. PMID- 21624691 TI - Pain management in the pediatric population. AB - Pain management is an important part of health care. During childhood, the prevalence of pain depends on the child's ongoing health status. Most healthy children receive more than 20 immunizations before 2 years of age. If the child is born with health concerns and is required to spend time in the neonatal intensive care unit, the number of painful encounters can reach into the hundreds. To optimally treat children with pain, nurses must realize that appropriately assessing and treating pain in children is a necessary part of their care. PMID- 21624692 TI - The 2010 American Heart Association guidelines for cardiopulmonary resuscitation and emergency cardiac care: an overview of the changes to pediatric basic and advanced life support. AB - The American Heart Association (AHA) has a strong commitment to implementing scientific research-based interventions for cardiopulmonary resuscitation and emergency cardiovascular care. This article presents the 2010 AHA major guideline changes to pediatric basic life support (BLS) and pediatric advanced life support (PALS) and the rationale for the changes. The following topics are covered in this article: (1) current understanding of cardiac arrest in the pediatric population, (2) major changes in pediatric BLS, and (3) major changes in PALS. PMID- 21624693 TI - Young adults with risk factors for chronic disease: transition needs for survivors of childhood cancer. AB - This article includes a definition of transition, the current state of transition, a review of transition research, an overview of chronic disease in survivors of childhood cancer (SCC), and the transition of SCC. In addition, models of transition are discussed, and the barriers to transition as well as principles for successful transition are identified. PMID- 21624694 TI - Pediatric sepsis and multiorgan dysfunction syndrome: progress and continued challenges. AB - Sepsis, septic shock, systemic inflammatory response syndrome, and Multiorgan dysfunction syndrome (MODS) remain clinical challenges in pediatric critical care. Understanding of the immune response has increased greatly over the past decade, which has certainly increased the understanding of the pathophysiology and treatment of these conditions. The future promises more exciting discoveries as we understand cellular physiology, immunity, and host responses even better. This article reviews the current knowledge about sepsis and MODS in pediatric patients and discusses the best treatment modalities while highlighting the critical aspects of nursing care for this vulnerable population. PMID- 21624695 TI - Transcatheter device closure for atrial septal defects: safety, efficacy, complications, and costs. AB - Atrial septal defects (ASDs) have traditionally been repaired by surgical closure. Recently, transcatheter device closure has increasingly been used with excellent results. Comparative research evaluating long-term outcomes of transcatheter technique data reveal significantly fewer complications and shorter hospital stays than those reported for surgical repairs. This article reviews relevant literature comparing safety and efficacy, costs, and complications of transcatheter device procedures with surgical closure of ASDs. PMID- 21624696 TI - Hematopoietic stem cell transplantation in children. AB - Medical and nursing care of the hematopoietic stem cell transplantation (HSCT) recipient are complex because of the pathophysiology, HSCT process, pre-HSCT conditioning regimens, numerous medications and therapies, acute and chronic complications, adverse effects, resources involved, and environmental considerations. The HSCT process and therapies may affect any body system, requiring proficient and prioritized nursing care, possibly in an intensive care setting. Understanding the timing of potential adverse effects and complications based on engraftment will help provide competent, high-acuity care. Although autogenic and allogeneic HSCT are curative treatment options, there are numerous morbidity and/or mortality risks throughout the HSCT journey. PMID- 21624697 TI - Ethical considerations in pediatric critical care research. AB - Pediatric critical care nurses are exposed to research in the critical care environment on a routine basis and should be knowledgeable about the ethical considerations inherent in this process. The following discussion includes information that centers on the ethical issues of conducting research with children. First, children as a vulnerable population is explored, followed by selected ethical principles that pertain to research, the role of the technological imperative in research, the process of informed consent, and finally, nursing considerations. PMID- 21624698 TI - Superficial doses in breast cancer radiotherapy using conventional and IMRT techniques: a film-based phantom study. AB - BACKGROUND AND PURPOSE: Superficial doses in radiotherapy are affected by the treatment technique. The implications for breast cancer treatments were evaluated. MATERIAL AND METHODS: Four treatment techniques relevant for breast cancer irradiation were evaluated; tangential standard, tangential IMRT, 7-field IMRT (arc-like field arrangement) and hybrid IMRT (an IMRT plan mixed with non modulated fields). Only 6MV photons were used. GafChromic EBT film was used for dose measurements at the surface, in the skin (0-5mm depth) and in the superficial parts of CTV (5-10mm depth) of an anthropomorphic thorax phantom. RESULTS: Only small differences in superficial doses were observed between tangential standard and tangential IMRT. Compared to the tangential standard plan, the surface and skin doses were reduced with the 7-field IMRT plan, on average by 20% and 5%, respectively, while hybrid IMRT reduced the surface and skin doses medially (by 44% and 8%, respectively) and increased the surface and skin dose laterally (by 40% and 15%, respectively). Minimum superficial CTV doses varied between regions, but were mainly between 90% and 95% of the target dose for all plans, only the hybrid IMRT plan resulted in a region with minimum dose below 90%. CONCLUSIONS: Compared to tangential irradiation, skin sparing was achieved by the 7-field IMRT plan. The minimum dose in the superficial parts of the CTV was below 95% of the target dose for all plans investigated. PMID- 21624699 TI - Can we decrease the skin reaction in breast cancer patients using hyaluronic acid during radiation therapy? Results of phase III randomised trial. AB - PURPOSE: Radio-induced early skin reactions still remain a clinical challenge. Preliminary results with Hyaluronic acid, one of the most recent topical products used in this indication are proving interesting. To evaluate the efficacy of Hyaluronic acid compared to placebo. MATERIAL AND METHODS: Breast cancer patients with grade 1-2 radio-induced dermatitis during postoperative radiotherapy were eligible. They were randomised to receive either hyaluronic acid (A) or a simple emollient (B). The primary endpoint was the clinical evaluation of the erythema (success versus failure). Secondary endpoints were the evaluation of skin colorimetry, pain, and quality of life. RESULTS: Two-hundred patients were enrolled (A=99, B=101). Ninety-five patients per treatment arm could be evaluated. Failures occurred in 23 patients (24%) in the hyaluronic acid arm, and 32 (34%) in the emollient arm (p=0.15). Seventy-three patients (36.5%) prematurely stopped the treatment without any ensuing difference between the two arms. Body mass index and the size of the epithelitis were both independently associated with the failure of the local treatment. The relative reduction of colorimetric levels was 20% in the hyaluronic acid group, and 13% in the emollient group (p=0.46). Concerning the quality of life assessment, there was a trend towards a lower level of pain in patients receiving hyaluronic acid (p=0.053). CONCLUSIONS: The present study showed no significant difference between hyaluronic acid and simple emollient in the treatment of acute radio induced dermatitis. There was however a trend towards an improvement in both pain level and skin colorimetry. PMID- 21624700 TI - Ex vivo evaluation of radical sun protection factor in popular sunscreens with antioxidants. AB - BACKGROUND: UVA induces tissue damage via the production of radical oxygen species. Adding antioxidants to UV filters in sunscreens is a novel photoprotective strategy. The topical application of antioxidants in sunscreen can potentially neutralize the UVA-induced free radicals. OBJECTIVES: We sought to assess the degree of free radical protection offered by sunscreens with antioxidants and attempted to differentiate the contribution of free radical protection from that of the UV filters. METHOD: Twelve sunscreen products were purchased. The degree of UVA protection (UVA-PF) was measured via an in vitro assay according to a European guideline (Colipa). In addition, an electron spin resonance (ESR) spectroscopy-based assay was used to measure the radical skin protection factor (RSF) and antioxidant power (AP) of each product. RESULTS: The sun protection factor (SPF) values of the sunscreens ranged from 15 to 55, and the UVA-PF values ranged from 2.4 to 28.2. The RSF values ranged from 2.4 to 27.1. There is a high correlation between RSF and UVA-PF. The AP values for nearly all the products were 0, and two products (#4 and #9) had very low AP values of 16 and 12, respectively. LIMITATIONS: The study only evaluated a small number of sunscreen products, and only ex vivo and in vitro methods were used to assess the products. CONCLUSIONS: The idea of combining UV filters with antioxidants is appealing. Current sunscreen products on the market offer free radical protection, but the majority of the radical protection is from UV filters rather than antioxidants. PMID- 21624701 TI - Risk of squamous cell carcinoma in junctional epidermolysis bullosa, non-Herlitz type: report of 7 cases and a review of the literature. AB - BACKGROUND: Squamous cell carcinoma (SCC) is the most severe complication and most common cause of death in patients with recessive dystrophic epidermolysis bullosa. The risk of developing SCC among patients with junctional epidermolysis bullosa (JEB) is unclear from the literature; however, in our center we noticed an unexpected number of SCCs among adult patients with JEB. OBJECTIVE: To review all documented patients with JEB in whom an SCC developed, both from our epidermolysis bullosa (EB) center and those reported in the literature. METHODS: A search in our EB registry documenting all JEB patients visiting our EB referral center from 1990 through 2010 revealed 7 JEB patients who developed 1 or more SCCs. A systematic literature search revealed 8 relevant articles documenting a total of 7 patients who developed an SCC. RESULTS: In our EB registry we found 7 patients with JEB who developed an SCC; these were all adults classified with non Herlitz type JEB. The frequency of developing an SCC among adult JEB patients (n = 28) in our center was therefore 25%. In the literature, we found 7 case reports of JEB complicated by SCC (also classified as JEB, non-Herlitz type), bringing the total number of documented cases to 14. The first SCC in JEB patients developed at an average age of 50 years (median, 52 years; range, 28-70 years). In 9 of 14 cases, multiple primary SCCs occurred, with a total of 45 SCCs. The SCCs are most often located on the lower extremities, in areas of chronic blistering, long-standing erosions, or atrophic scarring. Three patients (21%) developed metastases and died on average 8.9 years after diagnosis of the initial SCC. LIMITATIONS: This study was retrospective and the statistical analyses were based on a small number of patients. CONCLUSIONS: From their third decade, adult patients with JEB have an increased risk (1:4) of developing SCC on their lower extremities. The SCCs have a high recurrence rate and follow an aggressive course that results in death in 1 of 5 patients. We recommend annual checks of all JEB patients for SCC starting at 25 years of age. PMID- 21624702 TI - A window on professionalism in the emergency department through medical student narratives. AB - STUDY OBJECTIVE: We use medical students' reflections to deepen understanding of professionalism in the emergency department (ED), including the ideals that students wish to model and the lapses they hope to avoid. METHODS: Fourth-year students in a mandatory ED clerkship were required to write 2 narrative reflections during the month. The authors conducted a qualitative analysis to determine professionalism themes. RESULTS: Sixty-one of 150 student reflections contained professionalism themes. Positive behaviors included compassion, tension between respecting diversity and respecting other core values, and balance between patient-centered care and effective care. In addition, the students wrote about commitment to excellent medical care and ethical principles. Many students struggled with how to deal with patients who might be "drug seekers" and when to prescribe narcotics. They were concerned about the balance of compassion, often noting differences in the patients' backgrounds compared with their own. On the other hand, many students observed unprofessional behaviors. Particularly concerning were the absence of compassion, physicians lying, and a lack of teamwork. Students reflected on how their own professional behavior could improve according to their experiences in the ED. CONCLUSION: Students' reflective narratives are a rich source of information about good professional behavior, as well as threats to professionalism. Their experiences shaped the students' perceptions of emergency medicine and its values. Such reflections may constitute an important resource for faculty, student, and resident development. PMID- 21624703 TI - Variables associated with discordance between emergency physician and neurologist diagnoses of transient ischemic attacks in the emergency department. AB - STUDY OBJECTIVE: Transient ischemic attack is a common clinical diagnosis in emergency department (ED) patients with acute neurologic complaints. Accurate diagnosis of transient ischemic attack is essential to help guide evaluation and avoid treatment delays. We seek to determine the prevalence of discordant diagnosis for patients receiving an ED diagnosis of transient ischemic attack compared with neurologist final diagnosis. Secondary goals are to evaluate the influence of atypical transient ischemic attack symptoms, the ABCD2 score, and emergency physician experience on discordant diagnoses. METHODS: We performed a retrospective cohort study evaluating all ED patients receiving a diagnosis of transient ischemic attack during a 4-year period. The emergency physician diagnosis was compared with that of the neurologist. The neurologist's final diagnosis was considered the criterion standard diagnosis. Subject demographic and clinical information was collected with a structured instrument. The following atypical symptoms present at the ED evaluation were evaluated with logistic regression: headache, tingling, involuntary movement, seeing flashing lights or wavy lines, dizziness, confusion, incontinence, and ABCD2 score of 4 or greater. Bivariate analysis was used to evaluate the influence of emergency physician experience (<=6 years versus >6 years) on discordant diagnosis. Odds ratios (ORs) and proportions are reported with 95% confidence intervals (CIs), interquartile range was used where appropriate. RESULTS: We evaluated 436 subjects, of whom 7 were excluded, allowing 429 subjects for evaluation. Of these individuals, 156 (36%; 95% CI 32% to 41%) received a discordant diagnosis. The median emergency physician time in clinical practice was 6 years (interquartile range 2 to 12 years). Features associated with a discordant transient ischemic attack diagnosis included headache (OR 2.52; 95% CI 1.59 to 3.99), involuntary movement (OR 3.19; 95% CI 1.35 to 7.54), and dizziness (OR 1.92; 95% CI 1.22 to 3.02). Incontinence, confusion, and seeing wavy lines or flashing lights were not significantly associated with a discordant diagnosis. Patients with tingling and a high ABCD2 score had an increased odds of concordant transient ischemic attack diagnosis (OR 0.54, 95% CI 0.32 to 0.92; OR 0.53, 95% CI 0.35 to 0.82, respectively). CONCLUSION: Discordant diagnoses between emergency physicians and neurologists were observed in 36% of patients. The presence of headache, involuntary movement, and dizziness predicted discordant diagnoses, whereas the presence of tingling and an increased ABCD2 score predicted concordant transient ischemic attack diagnosis. PMID- 21624704 TI - Sutton's Law need not apply. PMID- 21624705 TI - Evidence-based emergency medicine. Are tissue adhesives an acceptable alternative for simple lacerations? PMID- 21624706 TI - [Cerebral reorganization in chronic low back pain and neurostimulation to improve motor control]. AB - Chronic recurrent pain results in brain reorganization for cortical sensory and motor representation of muscles. This review supports the hypothesis that maladaptive plasticity in chronic low back pain patients could be associated with disorders of volitional activation of trunk/pelvis muscles and alterations of their anticipatory motor patterns for postural control. This is actually observed for the transversus abdominis muscle, an abdominal muscle that stabilizes the lumbosacral spine against external and internal postural perturbations. Indeed, voluntary activation of this muscle is more difficult in low back pain and a posterolateral translation of its cortical motor representation is observed in association with a delay of its anticipatory activation (usually observed in any focal limb movement). During physiotherapy, low back pain patients are trained to hollowing for contracting the transversus abdominis muscle, which normalizes both activation delays and cortical motor representation. This motor rehabilitation could be potentiated by peripheral repetitive magnetic stimulation directly over the nerves/muscles. This review introduces the magnetic stimulation technique and proposes to combine peripheral theta-burst stimulation to conventional therapy for increasing the functional gain. This may facilitate the activation of sensory afferents and modulate cortical plasticity to improve (or even re-activate) the sensorimotor control and possibly influence pain. Peripheral magnetic stimulation as an ancillary treatment adjuvant for promoting motor control in low back pain might also be relevant in any other neurological or musculoskeletal disorder with underlying maladaptive brain reorganization. PMID- 21624707 TI - [Effects of unilateral fatigue of triceps surae on undisturbed stance]. AB - OBJECTIVE: Fatigue of lower limb presents various impacts on postural control. The purpose of this study was to explore the effects of a unilateral lower-limb muscle fatigue on undisturbed stance. METHODS: Nine healthy men performed until exhaustion a fatiguing exercise of the limb extensor muscles of one leg (exercised leg). Just before and after the fatiguing exercise, postural sway measures (centre of pressure) were recorded simultaneously from a double force platform and an in-shoe pressure system in quiet standing conditions. Maximal plantar pressure was computed from each sensor of the in-shoe system, before being averaged for the medial and lateral heel, the external mid-foot, the external and internal foreparts. RESULTS: In undisturbed stance, the fatiguing exercise induced postural destabilisation for the non-exercised leg (P < 0.001). Changes in antero-posterior mean position of the exercised leg were also observed reflecting a mean loading in a more rear foot stance (P < 0.01). The in-shoe system parameters revealed for the exercised foot, an increase of the maximal pressure value for the heel region (P < 0.05) and a decrease for the mid-foot and forepart (P < 0.05). For the non-exercised foot, the maximal pressure decreased only in forepart (P < 0.05). CONCLUSIONS: Unilateral triceps surae fatigue revealed an immediate destabilisation of undisturbed stance and the observed postural strategy appears similar to these developed by patients who presented pain pathology and/or sensory deficits in lower limb. PMID- 21624708 TI - Changes in the cutaneous silent period by paired stimulation. AB - OBJECTIVE: The cutaneous silent period (CSP) corresponds to the inhibition of motor neuronal activity that is induced by electrical cutaneous stimulation. This motor neuronal inhibition might be useful as a therapeutic strategy for modulating the excitability of motor neurons. Therefore, we investigated the CSP changes that can be observed using the paired-stimulation method. METHODS: Fifteen healthy adults were recruited. The digital cutaneous nerve of the right index finger was stimulated, and the CSP was recorded at the right thenar muscle. During the stimulation, contraction of the opposing right thumb and third finger was maintained at 20% of maximal voluntary contraction. A single stimulation was applied at the right index finger, and the duration and latency of the CSP (CSP1) was recorded. Paired electrical stimulations were then delivered with 60-, 80-, 100-, 120-, 140-, 160-, 180-, and 200-ms interstimulus intervals (ISI), and the latency and duration of a second CSP (CSP2) was measured and compared with that for the single stimulation. RESULTS: The CSP2 onset latencies were delayed in the 60-, 80-, and 100-ms ISI when compared to CSP1. CSP2 durations were shorter in the 60-, 80-, and 100-ms ISI. No significant differences in the latencies and durations between CSP1 and CSP2 were observed for ISI durations greater than 120 ms. CONCLUSIONS: We found that repetitive electrical stimulation changed the latency and duration of the CSP. These results suggest that the refractory period of the spinal inhibitory circuit in CSP is less than 100 ms. PMID- 21624709 TI - Cortical excitability of amyotrophic lateral sclerosis: transcranial magnetic stimulation study. AB - OBJECTIVE: The primary purpose of this study was to provide insight into the central changes that occur in amyotrophic lateral sclerosis (ALS) with a view to understanding how these could contribute to symptoms. MATERIAL AND METHODS: Seventeen patients with definite ALS and 17 control healthy volunteers were included in the study. Clinical examination, amyotrophic lateral sclerosis severity score (ALSSS) and TMS investigations including measurement of resting and active motor threshold (RMT and AMT), motor evoked potential (MEP), input output curve, contralateral silent period, and transcallosal inhibition (CSP and TI, postulated markers of GABAb function) were measured for each participant. RESULTS: There were no significant differences in RMT or AMT in either hemisphere between patients and the control group. Despite this there was a significant negative correlation between ALSSS and RMT and AMT meaning that increased severity was associated with higher thresholds. MEPs were significantly smaller in ALS patients in comparison to the control group (P = 0.03). There was a significant decrease in the slope of the I/O relationship of MEP amplitude to TMS intensity in patients group in comparison to controls. ALS patients had a significant prolongation of CSP and TI for both hemispheres. There was a tendency for a significant negative correlation between left TI and ALSSS (P = 0.051). CONCLUSION: Measurements of cortical motor excitatory changes in ALS confirm the presence of corticospinal hypoexcitability. Additionally we found increased excitability of presumed intracortical GABAb circuits that correlated with the severity of ALS. We postulate that the disease results in an imbalance between excitation and inhibition in the cortex that can contribute to clinical symptoms. PMID- 21624710 TI - Absent median nerve P14 far-field somatosensory evoked potential with persistent tibial nerve P30 component in a patient with ischemic pontine lesion. AB - In a patient with an ischemic lesion of the right paramedian region of the pons, somatosensory evoked potential (SEP) recording to median nerve stimulation showed an absent P14 response with still preserved P13 and N18 potentials. The tibial nerve P30 and N33 SEP components were normal. Our results suggest that the median nerve P14 potential, absent in our patient, has a different origin from the tibial nerve P30 response, normal in the present case. PMID- 21624711 TI - Block of the hERG channel by bupivacaine: Electrophysiological and modeling insights towards stereochemical optimization. AB - The hERG voltage-gated potassium channel mediates the cardiac I(Kr) current, which is crucial for the duration of the cardiac action potential. Undesired block of the channel by certain drugs may prolong the QT interval and increase the risk of malignant ventricular arrhythmias. Although the molecular determinants of hERG block have been intensively studied, not much is known about its stereoselectivity. Levo-(S)-bupivacaine was the first drug reported to have a higher affinity to block hERG than its enantiomer. This study strives to understand the principles underlying the stereoselectivity of bupivacaine block with the help of mutagenesis analyses and molecular modeling simulations. Electrophysiological measurements of mutated hERG channels allowed for the identification of residues involved in bupivacaine binding and stereoselectivity. Docking and molecular mechanics simulations for both enantiomers of bupivacaine and terfenadine (a non-stereoselective blocker) were performed inside an open state model of the hERG channel. The predicted binding modes enabled a clear depiction of ligand-protein interactions. Estimated binding affinities for both enantiomers were consistent with electrophysiological measurements. A similar computational procedure was applied to bupivacaine enantiomers towards two mutated hERG channels (Tyr652Ala and Phe656Ala). This study confirmed, at the molecular level, that bupivacaine stereoselectively binds the hERG channel. These results help to lay the foundation for structural guidelines to optimize the cardiotoxic profile of drug candidates in silico. PMID- 21624712 TI - Synthesis of new chalcone derivatives bearing 2,4-thiazolidinedione and benzoic acid moieties as potential anti-bacterial agents. AB - A series of chalcone derivatives bearing the 2,4-thiazolidinedione and benzoic acid moieties (8a-s) were synthesized, characterized, and evaluated for their anti-bacterial activity. Among the tested compounds, the most effective were 8a, 8h, 8k, 8n and 8q with MIC value in the range of 0.5-4 MUg/mL against six Gram positive bacteria (including multidrug-resistant clinical isolates). None of the compounds exhibited any activity against the Gram-negative bacteria Escherichia coli 1356 and E. coli 1682 at 64 MUg/mL. PMID- 21624713 TI - Dedication: Gary Purdue. PMID- 21624714 TI - Foreword. Burn rehabilitation. PMID- 21624715 TI - Preface: Burn rehabilitation. PMID- 21624716 TI - Acute assessment and management of burn injuries. AB - Burns are ubiquitous injuries in modern society, with virtually all adults having sustained a burn at some point in their lives. The skin is the largest organ of the body, basically functioning to protect self from non-self. Burn injury to the skin is painful, resource-intensive, and often associated with scarring, contracture formation, and long-term disability. Larger burns are associated with morbidity and mortality disproportionate to their initial appearance. Electrical and chemical burns are less common injuries but are often associated with significant morbidity. PMID- 21624717 TI - Burn wound care. AB - The goals of burn wound care are removal of nonviable tissue, prevention of infection, and facilitation of wound healing, while controlling pain and maximizing outcome. This article reviews the basic pathophysiology of burn wounds; describes the evaluation of the depth, location, and extent of the wound; and discusses the myriad of wound care products on the market including their strengths and weaknesses. This article guides the reader through wound assessment and designing the appropriate treatment plan. PMID- 21624718 TI - Positioning, splinting, and contracture management. AB - Whether a patient with burn injury is an adult or child, contracture management should be the primary focus of burn rehabilitation throughout the continuum of care. Positioning and splinting are crucial components of a comprehensive burn rehabilitation program that emphasizes contracture prevention. The emphasis of these devices throughout the phases of rehabilitation fluctuates to meet the changing needs of patients with burn injury. Early, effective, and consistent use of positioning devices and splints is recommended for successful management of burn scar contracture. PMID- 21624719 TI - Hand burns. AB - Burn rehabilitation cannot be reviewed without a significant focus on the hand. Although the surface area of the hand is only 3%, the functional consequences cause extreme impairment. A comprehensive team approach from initial evaluation through long-term follow up is essential to maximize the functional outcome in this population. PMID- 21624720 TI - Neurologic and musculoskeletal complications of burn injuries. AB - As more people survive burn injuries, there is an increasing focus on managing the complications of burn injuries with the ultimate goal of improving survivors' quality of life. Musculoskeletal and neurologic sequelae are significant complications of burn injury. Electrical injury is a subcategory of burns with multiple musculoskeletal and neurologic complications. Knowledge of these complications helps clinicians provide optimal long-term care for burn survivors and enables survivors to attain maximal recovery. PMID- 21624721 TI - Prosthetic management of the burn amputation. AB - Amputations as a result of thermal or electrical burns present a unique challenge that requires a coordinated effort by the rehabilitation team. Complications of surface and deep tissue injury require adjustment to the more standard approaches to prosthetic provision. Accommodations for the complications of joint contracture, skin adhesions, sensation compromise, skin grafts, and muscle flaps must be made by the treating clinicians. Differences in the treatment timelines, socket designs, material options, and component choices are discussed to familiarize the team for optimization of function in this difficult treatment scenario. PMID- 21624722 TI - Hypertrophic scar. AB - Hypertrophic scars are common complications of burn injury and other soft tissue injuries. Excessive extracellular matrix combined with inadequate remodeling of scar tissue results in an aesthetically and functionally unsatisfactory, painful, pruritic scar that can impair function. Treatment options are available to rehabilitation practitioners, but none are entirely satisfactory. An interdisciplinary clinical program is necessary for best outcomes. Challenges to be met by the rehabilitation community include research into the quantification of burn scar measurement, the effects of mechanical forces on wound healing and scar management, and the best combination of surgical, pharmacologic, and therapy interventions to maximize outcome from reconstructive procedures. PMID- 21624723 TI - Burn reconstruction. AB - The evolving shift in focus of burn care and research toward optimizing the long term outcomes of persons with burn injuries has certainly increased the emphasis on burn reconstruction. There are an increasing number of persons surviving extensive injury who may have long-term reconstructive needs. Burn reconstruction, just as acute burn care, requires a coordinated team approach from initial consultation through recovery and rehabilitation. Clearly, in the future, one can expect evolution in surgical techniques and technologies that can improve the function and appearance of persons with burn injury. PMID- 21624724 TI - Psychosocial recovery, pain, and itch after burn injuries. AB - Burn patients face complex rehabilitation issues including physical, emotional, social, and vocational challenges. Problems with anxiety, depression, sleep, pruritis, and body image can affect the burn patient's ability to return to an acceptable quality of life. Burn patients require not only help in the hospital setting but also expert care as an outpatient from vocational counselors, social workers, physical and occupational therapists, psychologists, professionally monitored support groups, and peer counselors. This article reviews the nonpharmacologic treatments available and discusses their scope and limitations. More research needs to focus on treatment options and prevention of long term issues. PMID- 21624725 TI - Exercise following burn injury. AB - Fatigue is a major barrier to recovery for burned individuals. Studies indicate that a slow return to normal or near-normal muscle strength is the natural course of recovery. With no special interventions, other than the "usual care" tailored to the needs of the individual, postburn patients will make gradual improvement in strength and aerobic capacity. Using the principle of initial condition (the worse the initial condition, the greater the response to exercise intervention) the authors outline an augmented exercise program that should result in a robust improvement in aerobic capacity. PMID- 21624726 TI - Community integration outcome after burn injury. AB - It is important to focus on community integration, including return to work and school, early during treatment after burn injuries. A careful analysis of the potential barriers to return to activities can help focus a treatment team and provide appropriate support for a return to work or school plan. Psychological intervention is often an important component of a return to work or school plan. Vocational rehabilitation counselors and school reentry coordinators are valuable assets to coordinating with a treatment team and communicating with a workplace or school. A successful return to work or school is often achieved with a coordinated and supportive approach. PMID- 21624727 TI - Geochronology of anthropogenic radionuclides in Ribeira Bay sediments, Rio de Janeiro, Brazil. AB - Ribeira Bay is located approximately 130 km south of the city of Rio de Janeiro and receives discharges of liquid effluent from the Angra dos Reis nuclear power plant (NPP) site, where two pressurized water reactors are located. To test whether the presence of anthropogenic radionuclides in sediments in Ribeira Bay could be correlated to the NPP operations, we sampled seven sediment cores and determined accumulation rates and chronologies. Only one sediment core did not exhibit a superficial mixing layer; this sample was used for dating purposes. Cesium-137 and (207)Bi were observed in this sediment profile, but their presence was associated with atmospheric fall-out rather than the nearby NPP. The exponential decay of (210)Pb concentration with sediment layer depth was verified below a superficial mixing layer for all other sediment cores. Calculated accumulation rates ranged from 1.2 mm y(-1) in the inner bay to 6.2 mm y(-1) close to its entrance. PMID- 21624728 TI - Informing health? Negotiating the logics of choice and care in everyday practices of 'healthy living'. AB - This paper reports on a qualitative study examining everyday practices of healthy living (HL). Forty-four semi-structured interviews were undertaken with Canadian and UK citizens, aged 45 - 70, in April-May 2010. The research sits within the now substantial literature concerned with how health information is mediated, both by people and technologies, and employed in the context of 'good' health citizenship. Throughout this work, notions of 'choice' and 'empowerment' have been interrogated, theoretically and empirically, to reveal both the knowledge/power relationships integral to 'informing' processes and the shifting relationship between information and care in contemporary health encounters. In this paper, we analyse how people make sense of what it means to live healthily and how they know if they are doing so by focussing on three ways in which study participants become informed about healthy living: through their engagement with universal HL messages, through their own information searches, and through their attempts to measure their 'healthiness'. Following Mol's (2008) critique of the "logic of choice" in contemporary healthcare, we understand healthy living as a "situation of choice" where complex problems are framed as simple matters of choice and where information and technologies are understood as neutral aids to decision-making in support of 'correct' choices. Our analysis builds on and extends Mol's work by exploring how participants negotiate between this "logic of choice" and her alternative "logic of care" in their accounts of everyday HL informing practices and how the two logics "interfere" with one another. These accounts show resistance to the logic of choice through 'calls for care' but they also show clearly how the disciplining logic of choice works to (re)present such calls for care as failed attempts at healthy living, undermining the very practices the logic of choice seeks to encourage. PMID- 21624729 TI - Feasibility, acceptability, and initial efficacy of a knowledge-contact program to reduce mental illness stigma and improve mental health literacy in adolescents. AB - The purpose of this school-based cluster-randomized trial was to determine the initial acceptability, feasibility, and efficacy of an existing community-based intervention, In Our Own Voice, in a sample of US adolescent girls aged 13-17 years (n = 156). In Our Own Voice is a knowledge-contact intervention that provides knowledge about mental illness to improve mental health literacy and facilitates intergroup contact with persons with mental illness as a means to reduce mental illness stigma. This longitudinal study was set in two public high schools located in a southern urban community of the U.S. Outcomes included measures of mental illness stigma and mental health literacy. Findings support the acceptability and feasibility of the intervention for adolescents who enrolled in the study. Findings to support the efficacy of In Our Own Voice to reduce stigma and improve mental health literacy are mixed. The intervention did not reduce mental illness stigma or improve mental health literacy at one week follow up. The intervention did not reduce mental illness stigma at 4 and 8 weeks follow up. The intervention did improve mental health literacy at 4 and 8 weeks follow up. Previous studies have assessed the preliminary efficacy In Our Own Voice among young adults; rarely has In Our Own Voice been investigated longitudinally and with adolescents in the United States. This study provides initial data on the effects of In Our Own Voice for this population and can be used to further adapt the intervention for adolescents. PMID- 21624731 TI - The year in quality of care in heart failure. PMID- 21624732 TI - Human serum albumin in the clinical syndrome of heart failure. AB - Hypoalbuminemia is common in patients with heart failure, and this condition becomes more prevalent with increasing age and illness. Hypoalbuminemia is thought to result mainly from malnutrition, inflammation and cachexia. Other causal factors include hemodilution, liver dysfunction, protein-losing enteropathy, increased transcapillary escape rate, and nephrotic syndrome. According to Starling's law, low plasma oncotic pressure related to hypoalbuminemia induces a fluid shift from the intravascular to the interstitial space, and there is now clinical evidence that hypoalbuminemia facilitates the onset of cardiogenic pulmonary edema. Hypoalbuminemia has emerged as an independent predictor of incident heart failure in end-stage renal disease and elderly patients. Recent data also suggest that hypoalbuminemia provides prognostic information incremental to the usual clinical and biochemical variables in patients with heart failure regardless of clinical presentation. The presence of hypoalbuminemia in patients with heart failure may have potential therapeutic consequence in clinical practice. If present, subclinical excess of fluid must be removed. A dietary survey should also be performed, and renutrition may be indicated. It is unknown whether targeted nutritional intervention and albumin administration confer benefits to hypoalbuminemic patients with heart failure, and further research is warranted in this setting. PMID- 21624733 TI - Risk of sudden death in end-stage hypertrophic cardiomyopathy. AB - BACKGROUND: It remains unclear whether end-stage hypertrophic cardiomyopathy (HCM) is associated with as high a rate of sudden death as occurs among HCM patients with preserved left ventricular (LV) systolic function. The purpose of this study was to evaluate the incidence of sudden death among patients with end stage HCM and to identify high-risk end-stage patients. METHODS AND RESULTS: A total of 490 consecutive patients with HCM, who were diagnosed and followed-up at our hospital, were analyzed retrospectively. End-stage HCM was defined by an LV ejection fraction <50% on echocardiography during follow-up. Among the 490 HCM patients, 43 patients (8.8%) were diagnosed as having end-stage HCM during a mean follow-up period of 12 +/- 7 years after the initial diagnosis. During a mean follow-up period of 5 +/- 3 years after progression to end-stage HCM, sudden death occurred in 21 of 43 patients (47%). Cox proportional hazards analysis identified syncope as an independent predictor of sudden death (hazard ratio = 6.15; 95% confidence interval, 2.40-15.75; P < .001). CONCLUSIONS: This study demonstrated that patients with end-stage HCM have a high incidence of sudden death. Therefore, it is suggested that an aggressive therapeutic strategy to counter sudden death should be considered for patients with end-stage HCM. PMID- 21624734 TI - Plasma TIMP-4 predicts left ventricular remodeling after acute myocardial infarction. AB - BACKGROUND: Alterations in the balance between matrix metalloproteinases and their endogenous tissue inhibitors (TIMPs) are associated with left ventricular (LV) remodeling after acute myocardial infarction (AMI). No relationships have been identified between TIMPs and serial postinfarction change in LV function. METHODS AND RESULTS: Plasma concentrations of TIMP-1, -2, -4 were measured at baseline (mean 46 h) and at 24 weeks in 100 patients (age 58.9 +/- 12 years, 77% male) admitted with AMI and LV dysfunction, with cardiac magnetic resonance imaging at each time point. TIMP-1 concentration was reduced, whereas TIMP-2 and 4 concentrations were elevated at baseline compared with a reference control population. TIMP-1 decreased and TIMP-2 increased significantly over time; there was an incremental trend in TIMP-4 concentration. Baseline TIMP-4 correlated with change in LV end-systolic volume index (?LVESVI; r = 0.24; P = .023) and change in LV end-diastolic volume index (?LVEDVI; r = 0.25; P = .015). ?TIMP-4 also correlated with ?LVESVI and with ?LVEDVI, as did ?TIMP-2. On multivariable analysis, baseline TIMP-4 concentration was an independent predictor of ?LVESVI. CONCLUSIONS: Plasma TIMP-4 concentration, measured early after AMI, may assist in the prediction of LV remodeling and therefore in the assessment of prognosis. Further study of the role of the TIMPs in the pathophysiology of postinfarction remodeling is warranted. PMID- 21624735 TI - Renal dysfunction is a stronger determinant of systemic neutrophil gelatinase associated lipocalin levels than myocardial dysfunction in systolic heart failure. AB - BACKGROUND: Neutrophil gelatinase-associated lipocalin (NGAL) is released by renal tubular cells in response to inflammation and injury. Recent studies have demonstrated that NGAL is up-regulated in cardiomyocytes within the failing myocardium. However, the overall relationship between systemic NGAL levels and myocardial structure and performance has not been established. METHODS AND RESULTS: We measured systemic NGAL levels in 130 subjects with chronic systolic heart failure (HF) and comprehensive echocardiographic evaluation, as well as 69 subjects with acute decompensated systolic HF and hemodynamic evaluation. In the chronic HF cohort, higher plasma NGAL levels were modestly associated with increasing age (r = 0.18; P = .035), higher New York Heart Association functional class (rank sums: P = .022) and impaired renal function (eGFR: r = -0.53; P < .0001; cystatin C: r = 0.60; P < .0001). Plasma NGAL levels were modestly associated with indices of diastolic dysfunction (mitral E/Ea: r = 0.27; P = .002; LAVi: r = 0.25; P = .011; tricuspid E/Ea: r = 0.20; P = .029), but not after adjustment for renal function (P > .10 for all). In Cox proportional hazards analysis, plasma NGAL predicted cardiac death or transplantation after adjustment for age, gender, left ventricular ejection fraction, and mitral E/Ea (hazard ratio 1.68, 95% confidence interval 1.08-2.57; P = .022), but not after adjustment for renal function (P = .83). In the acute HF cohort, we did not observe any relationship between NGAL and hemodynamic indices, but NGAL strongly correlated with renal function. CONCLUSIONS: Systemic NGAL levels are largely determined by underlying impairment of renal rather than myocardial function. Our findings did not support any prognostic significance or relationship between systemic NGAL levels and indices of cardiac structure and function after adjustment for underlying renal function. PMID- 21624736 TI - Amino-terminal pro-B-type natriuretic peptide (NT-proBNP) levels 3 months after myocardial infarction are more strongly associated with magnetic resonance determined ejection fraction than NTproBNP levels in the acute phase. AB - BACKGROUND: The relationship between levels of amino-terminal pro-B-type natriuretic peptide (NT-proBNP) and left ventricular function determined by magnetic resonance imaging (MRI) in ST-segment-elevation myocardial infarction (STEMI) is largely unknown. METHODS AND RESULTS: This was a substudy of the Norwegian Study on District Treatment of STEMI, in which patients received thrombolysis followed by early or late invasive strategy. NT-proBNP was measured at 3 days and 3 months after the myocardial infarction, and magnetic resonance imaging was performed after 3 months (n = 160). Log NT-proBNP levels at both time points were significantly associated with ejection fraction (EF) (r(2) values 0.25 and 0.42, respectively) as well as infarct size (r(2) values 0.38 and 0.47, respectively; P < .0001 for all, adjusted for confounders). Furthermore, receiver operating characteristic (ROC) curves used to analyze the ability of NT-proBNP to discriminate long-term low EF (<=40%) and large infarct size (>=15.7%), were significant at both time points (P < .001 for all). Pairwise comparison of the ROC curves showed a significantly better performance of NT-proBNP at 3 months compared with 3 days for discrimination of low EF (P = .023). CONCLUSION: Repeated measurements of NT-proBNP in STEMI patients showed that NT-proBNP levels at 3 months were more strongly associated with long-term EF and infarct size than NT-pro BNP levels after 3 days. Our data suggest that measurement of NT-proBNP 3 months after myocardial infarction is a better indicator of left ventricular function compared with NT-proBNP in the acute phase. PMID- 21624737 TI - Effectiveness of mechanical circulatory support in children with acute fulminant and persistent myocarditis. AB - BACKGROUND: Acute fulminant myocarditis is a life-threatening disease in children. A limited number of reports suggest that mechanical circulatory support (MCS) may be used to successfully bridge children with acute fulminant myocarditis to recovery or transplantation. We evaluated the effectiveness of MCS in children with myocarditis and identified risk factors associated with adverse outcomes. METHODS AND RESULTS: Between 2001 and 2009, 16 children were treated for myocarditis at our institution; each child received MCS provided by extracorporeal membrane oxygenation, ventricular assist device(s), or both. Of these patients, 75% (12/16) survived: 7 recovered ventricular function, and 5 underwent successful orthotopic heart transplantation. In patients who were bridged to recovery, mean left ventricular ejection fraction significantly improved from initiation to termination of MCS (20 +/- 9.3% to 62 +/- 5%; P = .0004). Viral pathogens were detected in 11 patients by polymerase chain reaction, and viral presence was associated with death or need for transplantation (P = .011). Upon histologic analysis, absence of viral infection and lack of myocardial inflammation were associated with recovery (P values .011 and .044, respectively). CONCLUSIONS: In children with acute fulminant and persistent myocarditis, MCS is a life-saving treatment strategy, particularly in the absence of viral infection. PMID- 21624738 TI - Torsion and dyssynchrony differences between chronically paced and non-paced heart failure patients. AB - BACKGROUND: Chronic right ventricular pacing may lead to left ventricular dyssynchrony, systolic dysfunction, remodeling, and heart failure. Cardiac mechanics may differ between paced and nonpaced heart failure patients, and their optimal treatment may also differ. METHODS AND RESULTS: Echocardiograms were analyzed using tissue Doppler imaging and speckle tracking echocardiography in 20 patients with chronic right ventricular pacing for complete heart block (RVP group), 29 nonpaced patients with different heart failure etiologies but ejection fractions similar to the RVP group (HF group), and 25 control subjects without pacemakers or heart failure (control group). Left ventricle volumes were smaller in RVP than HF (end-diastolic volume = 93.6 +/- 25.1 mL vs. 112.1 +/- 22.8 mL), but intraventricular longitudinal and radial dyssynchrony were similar. Dyssynchrony within the septum was greater (number of segments lengthening during systole = 1.9 +/- 1.7 vs. 0.9 +/- 1.8), systolic torsion was lower (6.2 +/- 7.3 degrees vs. 10.6 +/- 4.2 degrees ), untwisting was delayed (time from peak torsion to peak untwist rate = 188 +/- 141 ms vs. 102 +/- 73 ms), and apical rotation was reversed in more subjects (35% vs 0%) in RVP than HF groups (P < .05 for all). CONCLUSIONS: Intraventricular dyssynchrony was similar between RVP and HF groups with similar ejection fraction. However, RVP subjects had smaller ventricles, greater dyssynchrony within the septum, lower torsion, altered apical rotation, and delayed untwisting. PMID- 21624739 TI - The effects of depression on the course of functional limitations in patients with chronic heart failure. AB - BACKGROUND: This study examined whether depressive symptoms are associated with persistent functional limitations and severity classified according to the course of functional limitations in chronic heart failure (CHF) patients after discharge. METHODS AND RESULTS: The Performance Measure for Activities of Daily Living 8 (PMADL-8) was used to measure the course of functional limitations at 1, 3, and 5 months after discharge in a cohort of 148 patients. Depressive symptoms were assessed using the Hospital Anxiety and Depression Scale at 1 month after discharge. Repeated-measures logistic regression adjusting for potential confounders demonstrated that the depression groups had different persistent functional limitations (PMADL-8 scores >=20 at 1, 3, and 5 months after discharge [chi(2) = 5.3; P < .05]). Using cluster analysis, we identified 4 distinctive courses of functional limitations, and there was a graded relationship between the severity of the course and depressive symptoms (chi(2) = 26.1; P < .001). CONCLUSIONS: In this prospective study, depression was associated with poorer functional limitations in CHF patients after discharge. The findings of this study suggest that depression may be a treatment target for improving functional limitations in CHF patients during the recovery phase. PMID- 21624740 TI - Use of the aortoatrial continuity as means of providing left ventricular assist support without entering the ventricle: a feasibility study. AB - BACKGROUND: Left ventricular assist devices (LVADs) are increasingly used to treat patients with refractory heart failure. Current-generation LVADs have major limitations, including the need for open chest surgery, limiting their widespread use. We hypothesized that the aortoatrial continuity could be used as a unique anatomic vantage point for entirely percutaneous LVAD placement. METHODS AND RESULTS: Forty human autopsied hearts were examined to ascertain the presence and define the dimensions of the continuity between the posterior aortic wall and the left atrium. In all cases, a "septum" between the aorta and left atrium was identified. In 3 animal experiments, a custom mechanical shunt was deployed in the wall between the left atrium and noncoronary cusp. With continuous intracardiac ultrasound imaging, and at necropsy, there was no evidence of device dislodgement, pericardial effusion, or aortic or coronary artery trauma noted. CONCLUSIONS: It is feasible to use the wall between the aorta and left atrium as an access route for a potentially entirely percutaneous LVAD. Such a system obviates the need for accessing the left ventricle, minimizing complications. In the future, such devices may allow widespread treatment of heart failure, malignant cardiac arrhythmia, and severe aortic and mitral valvular disease. PMID- 21624741 TI - Baroreflex sensitivity impairment is associated with cardiac diastolic dysfunction in rats. AB - BACKGROUND: Studies have shown that the autonomic dysfunction accompanied by impaired baroreflex sensitivity was associated with higher mortality. However, the influence of decreased baroreflex sensitivity on cardiac function, especially in diastolic function, is not well understood. This study evaluated the morphofunctional changes associated with baroreflex impairment induced by chronic sinoaortic denervation (SAD). METHODS AND RESULTS: Animals were divided into sinoaortic denervation (SAD) and control (C) groups. Baroreflex sensitivity was evaluated by tachycardic and bradycardic responses, induced by vasoactive drugs. Cardiac function was studied by echocardiography and by left ventricle (LV) catheterization. LV collagen content and the expression of regulatory proteins involved in intracellular Ca(2+) homeostasis were quantified. Results showed higher LV mass in SAD versus C animals. Furthermore, an increase in deceleration time of E-wave in the SAD versus the C group (2.14 +/- 0.07 ms vs 1.78 +/- 0.03 ms) was observed. LV end-diastolic pressure was increased and the minimum dP/dt was decreased in the SAD versus the C group (12 +/- 1.5 mm Hg vs 5.3 +/- 0.2 mm Hg and 7,422 +/- 201 vs 4,999 +/- 345 mm Hg/s, respectively). SERCA/NCX ratio was lower in SAD than in control rats. The same was verified in SERCA/PLB ratio. CONCLUSIONS: The results suggest that baroreflex dysfunction is associated with cardiac diastolic dysfunction independently of the presence of other risk factors. PMID- 21624743 TI - Human exhaled breath analysis. AB - OBJECTIVE: To review the fast-developing topic of assessment of exhaled breath components to improve the diagnosis and monitoring of respiratory and systemic diseases. DATA SOURCES: Review of the literature available in monographs and journals. STUDY SELECTION: Articles and overviews on the broad spectrum of existing experimental and routinely applied methods to assess different aspects of human exhaled breath analysis were selected for presentation in this review. RESULTS: Exhaled breath constitutes more than 3,500 components, the bulk of which are volatile organic compounds in miniature quantities. Many of these characterize the functioning of the organism as a whole (systemic biomarkers), but some are related to processes taking place in the respiratory system and the airways in particular (lung biomarkers). Assessment of lung biomarkers has proven useful in airway inflammatory diseases. It involves direct measurement of gases such as nitric oxide and inflammatory indicators in exhaled breath condensate such as oxidative stress markers (eg, hydrogen peroxide and isoprostanes), nitric oxide derivatives (eg, nitrate and nitrates), arachidonic acid metabolites (eg, prostanoids, leukotrienes, and epoxides), adenosine, and cytokines. Integral approaches have also been suggested, such as exhaled breath temperature measurement and devices of the "electronic nose" type, which enable the capture of approaches have also been suggested, such as exhaled breath temperature measurementexhaled molecular fingerprints (breath prints). Technical factors related to standardization of the different techniques need to be resolved to reach the stage of routine applicability. CONCLUSIONS: Examination of exhaled breath has the potential to change the existing routine approaches in human medicine. The rapidly developing new analytical and computer technologies along with novel, unorthodox ideas are prerequisites for future advances in this field. PMID- 21624744 TI - Relationship between bronchial hyperreactivity and bronchodilation in patients with allergic rhinitis. AB - BACKGROUND: Allergic rhinitis (AR) may be considered a risk factor for the onset of asthma. Recently, it has been reported that forced expiratory flow between 25% and 75% of vital capacity (FEF(25%-75%)) may predict a positive response to bronchodilation test in asthmatic children. Moreover, bronchial hyperreactivity (BHR) is frequently detected in AR patients. OBJECTIVE: To evaluate the possible relationship between the response to bronchodilation test and methacholine challenge, also considering the FEF(25%-75%) values in a large group of patients with persistent allergic rhinitis. METHODS: Three hundred sixty-five consecutive AR patients were evaluated. Clinical examination, spirometry, methacholine bronchial challenge, and bronchodilation test were performed in all patients. RESULTS: Two hundred forty-one patients (66%) had "positive" results for bronchodilation test; FEF(25%-75%) was abnormal in 78 patients (21.4%), and 76 patients (20.8%) had severe BHR. An FEF(25%-75%) cutoff value of less than 58.5% of predicted may optimally (AUC 0.97) discriminate patients with both severe BHR and reversibility. CONCLUSION: This study confirms previous studies and increases the strength of the role of FEF(25%-75%) as a marker of early bronchial involvement in patients suffering from persistent allergic rhinitis. Moreover, an FEF(25%-75%) value less than 58.5% of predicted may suggest the co-existence of severe BHR and reversibility. PMID- 21624745 TI - High asthma prevalence and increased morbidity among rural children in a Medicaid cohort. AB - BACKGROUND: Urban children represent a group at high risk for asthma development and adverse asthma outcomes. Although rural children also encounter sociodemographic disparities that might be expected to worsen asthma, asthma in the rural United States is poorly studied. OBJECTIVES: To determine rural-urban differences in childhood asthma diagnosis and morbidity. METHODS: We studied a statewide population of 117,080 children continuously enrolled in Tennessee Medicaid from birth through the sixth year of life, using linked Tennessee Medicaid, vital records, and pharmacy claims databases to determine asthma diagnosis and residence. RESULTS: The cohort was 45% urban, 23% suburban, and 33% rural. Compared with urban children, rural children were more likely to be white, have a history of bronchiolitis, and have mothers who smoked. Eleven percent of urban, 12% of suburban, and 13% of rural children met study criteria for asthma diagnosis (adjusted odds ratio for rural children, 1.16; 95% confidence interval, 1.09-1.24; adjusted odds ratio for suburban children, 1.22; 95% confidence interval, 1.14-1.30; with urban as the referent; P < .001). Rural children had greater use of outpatient asthma care, whereas urban children had greater use of inhaled corticosteroids. Compared with urban children, rural children had fewer asthma emergency department visits but were hospitalized for asthma at similar rates and had similar use of asthma rescue medications. CONCLUSION: In this pediatric Medicaid population, rural children had increased asthma prevalence and similar asthma morbidity compared with urban children but differences in patterns of asthma care and resource use, suggesting that optimal interventions for asthma may differ in rural compared with urban populations. PMID- 21624746 TI - Association between genes encoding components of the IL-10/IL-0 receptor pathway and asthma in children. AB - BACKGROUND: Asthma is a chronic inflammation of the airways associated with recurrent symptoms that range from mild to debilitating. Interleukin-10 (IL-10) is a cytokine that displays pleiotropic effects in asthma and allergy. OBJECTIVE: To determine whether polymorphisms of IL-10/IL-10R pathway contribute to asthma susceptibility in Egyptian children. METHODS: The IL-10 (-1082G/A), IL-10R1 (G330R), and signal transducer and activator of transcription 3 (STAT3) rs2293452 single-nucleotide polymorphism (SNP) were genotyped in 110 atopic children with asthma, 110 non-atopic children with asthma, and 110 healthy children. Serum IL 10 and immunoglobulin E (IgE) levels were determined by enzyme-linked immunosorbent assay. RESULTS: A significant association was observed between the IL-10 polymorphism and asthma in both atopic (P = .03) and non-atopic asthma groups (P = .04). The genotype frequencies of IL-10R1 polymorphisms did not differ between all groups. We identified a significant association between STAT3 polymorphism and asthma susceptibility in atopic asthma (P < .001), whereas no such association was observed in the non-atopic asthma group (P = .9). No evidence of gene interactions was found. CONCLUSION: Polymorphisms of IL-10 and STAT3 may be useful as a new DNA-based diagnostic biomarker for identifying high risk children susceptible to asthma. PMID- 21624747 TI - Effects of aqueous extract of Echinodorus grandiflorus on the immune response in ovalbumin-induced pulmonary allergy. AB - BACKGROUND: Asthma is a disease characterized by intermittent obstruction of the airways and chronic inflammation that affects approximately 300 million people worldwide. The immune response in asthma is predominantly T(H)2, with high levels of total and allergen-specific IgE and bronchial eosinophilia. Asthma treatment is aimed at controlling the disease, and the drugs used currently have systemic adverse effects and generally are not effective in difficult-to-control cases. OBJECTIVE: To investigate the effect of aqueous extract of Echinodorus grandiflorus, a plant used in folk medicine for its diuretic and anti inflammatory properties, in a model of pulmonary allergy. METHODS: BALB/c mice were intraperitoneally sensitized and nasally challenged with ovalbumin. Aqueous extract and dexamethasone treatments (0.1 mL/d per mouse) were initiated on day 32 and concluded on day 40. Eight hours after the last challenge evaluations, of serum, bronchoalveolar lavage, and lung tissue were performed. RESULTS: Oral treatment with the extract markedly reduced the number of total cells and eosinophils in bronchoalveolar lavage. The eosinophil peroxidase activity in lung tissue, the levels of ovalbumin-specific IgE in serum, the levels of CCL11, and the gene expression of interleukin 4 and interleukin 13 in lung tissue were also lower after treatment. CONCLUSIONS: These results suggest that the aqueous extract of E grandiflorus is able to modulate allergic pulmonary inflammation and may be useful as a potential therapeutic agent for asthma. PMID- 21624748 TI - Prescriptions for self-injectable epinephrine in emergency department angioedema management. AB - BACKGROUND: Angioedema is often treated in the emergency department (ED). Few studies have evaluated self-injectable epinephrine (SIE) prescribing patterns for angioedema. OBJECTIVES: To describe presentation and management of ED patients with angioedema and determine factors associated with epinephrine administration, hospital admission and SIE prescription. METHODS: We conducted a retrospective cohort study of all ED patients with angioedema between January 2005 and December 2006. RESULTS: Of 63 patients, 39 (61.9%) were female. Median age was 49 years. Precipitating factors were identified in 36 (57.1%) patients. History of other allergic conditions was seen in 37 (58.7%) patients. Seventeen (27.0%) patients received epinephrine, 55 (87.3%) received antihistamines, and 51 (81.0%) received steroids. Epinephrine was administered more commonly in patients with edema of the tongue (risk ratio [RR], 5.28, 95% confidence interval [CI] 1.95-14.33, P = .0003), tightness/fullness of throat (RR, 3.31, 95% CI 1.62-6.76, P = .006), and dyspnea/wheeze (RR, 3.04, 95% CI 1.41-6.59, P = .005). Hospitalization was more common in patients with dyspnea/wheeze (P = .028) and allergic history (P = .006). Thirteen patients (22.0%) were discharged with SIE. An SIE prescription was associated with younger patients (median age, 26 years [interquartile range (IQR) 15-50] vs a median age 57.5 years [IQR 43-68], P = .004) and patients with throat tightness/fullness (RR, 4.2, 95% CI 1.8-9.8, P = .005). CONCLUSION: Patients with respiratory symptoms and allergic history were likely to be admitted. Epinephrine use was more frequent in patients with signs and symptoms of oropharyngeal edema. Younger patients and those with tightness/fullness of throat were likely to be prescribed SIE. Further studies are needed to determine who would benefit from epinephrine use and SIE prescription. PMID- 21624749 TI - Seafood allergy in children: a descriptive study. AB - BACKGROUND: Food allergy and seafood (fish, mollusk, and crustacean) consumption have increased considerably over the past 40 years. Seafood allergy is now a leading cause of anaphylaxis in both the United States and Australia. However, there is only limited published data describing the clinical presentation and management of seafood allergy. OBJECTIVES: To describe the characteristics of a large cohort of children with seafood allergy. METHODS: Using a retrospective chart review, we collected data on all children presenting to our Tertiary Allergy Service with an allergic reaction to seafood between 2006 and 2009. RESULTS: 167 children had a history of definite clinical reaction to seafood and/or positive food challenge (103 male, 62%). 94% had evidence of co-existent atopic disease. Prawn/shrimp was the most common seafood implicated. One-fifth presented with a history of anaphylaxis to seafood. Over 50% of crustacean allergic children could tolerate non-crustacean fish. Sensitization to other fish species was very common in fish-allergic children, with one third reporting clinical reactions to at least two species; 16% developed symptoms to fish vapours. In children with allergy to tuna and/or salmon, at least 21% were able to tolerate the fish in a tinned form. CONCLUSIONS: Seafood is a relatively common and important cause of food allergy in Australian children, presenting with a high rate of anaphylaxis. PMID- 21624750 TI - Changes in gene expression caused by insect venom immunotherapy responsible for the long-term protection of insect venom-allergic patients. AB - BACKGROUND: Insect venom immunotherapy (VIT) is the only causative treatment of insect venom allergy (IVA). The immunological mechanism(s) responsible for long term protection achieved by VIT are largely unknown. A better understanding is relevant for improving the diagnosis, prediction of anaphylaxis, and monitoring and simplifying treatment of IVA. OBJECTIVE: To find genes that are differentially expressed during the maintenance phase of VIT and after stopping, to get clues about the pathways involved in the long-term protective effect of immunotherapy. METHODS: Whole genome gene expression analysis was performed on RNA samples from 50 patients treated with VIT and 43 healthy controls. Patients were divided into three groups: (1) before the start of VIT; (2) on maintenance phase of VIT for at least 3 years still receiving injections; and (3) after VIT. RESULTS: Of all 48,804 probes present in the array, 48,773 transcripts had sufficient data for further analysis. The list of genes that were differentially expressed (at least log2 FC > 2; P < .05 corrected for multiple testing) during the maintenance phase of VIT as well as after successful VIT contains 89 entities. The function of these genes affects cell signaling, cell differentiation, and ion transport. CONCLUSION: This study shows that a group of genes is differentially expressed both during and after VIT in comparison with gene expression in patients before VIT. Although the results of this study should be confirmed prospectively, the relevance of these findings is supported by the fact that they are related to putative mechanisms of immunotherapy. PMID- 21624751 TI - The opposing role of two prostaglandin D2 receptors, DP and CRTH2, in human eosinophil migration. AB - BACKGROUND: Prostaglandin D2 (PGD2) regulates various immunological responses via two distinct PGD2 receptors, prostaglandin D receptor (DP), and chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2). Recent studies have demonstrated that PGD2 induces the migration of eosinophils through CRTH2. Although human eosinophils express both DP and CRTH2, it is unclear whether the function of DP is involved in eosinophil migration. OBJECTIVE: In this study, we investigated the roles of DP and CRTH2 in eosinophil migration by using selective agonists and antagonists. METHODS: Eosinophils were isolated from human subjects with mild eosinophilia by modified CD16-negative selection. After stimulation with or without DP receptor agonist, eosinophil migration was measured by Boyden chamber. The effect of DP agonists on CRTH2-induced eosinophil migration was studied in terms of CRTH2 expression, Ca2+ mobilization, ERK/MAPK phosphorylation, and cyclic AMP (cAMP) production. RESULTS: Treatment with DP agonists inhibited CRTH2-induced chemotaxis of eosinophils. Furthermore, we showed that DP agonists enhanced cAMP production in CRTH2 agonist stimulation without increasing CRTH2 expression. DP mediates eosinophils through the elevation of intracellular cAMP production but does not change CRTH2 expression. CONCLUSION: Taken together, the balance between DP and CRTH2 could influence the degree of PGD2-induced eosinophil migration and DP agonist might regulate eosinophil activation. PMID- 21624752 TI - Intermittent or daily montelukast versus placebo for episodic asthma in children. AB - BACKGROUND: No standard, optimal treatment exists for severe intermittent (ie, episodic) asthma in children. However, evidence suggests that both daily and episode-driven montelukast are effective for this phenotype. OBJECTIVE: To assess the regimen-related efficacy of montelukast in treating pediatric episodic asthma. METHODS: A multicenter, randomized, double-blind, double-dummy, parallel group, 52-week study was performed in children 6 months to 5 years of age comparing placebo with two regimens of montelukast 4 mg: (1) daily; or (2) episode-driven for 12 days beginning with signs/symptoms consistent with imminent cold or breathing problem. The main outcome measure was the number of asthma episodes (symptoms requiring treatment) culminating in an asthma attack (symptoms requiring physician visit, emergency room visit, corticosteroids, or hospitalization). RESULTS: Five hundred eighty-nine patients were randomized to daily montelukast, 591 to intermittent montelukast, and 591 to placebo. Compared with placebo, no significant difference was seen between daily montelukast (P = .510) or intermittent montelukast (P = .884) in the number of asthma episodes culminating in an asthma attack over 1 year. Daily montelukast reduced symptoms over the 12-day treatment period of asthma episodes compared with placebo (P = .045). Beta-agonist use was reduced with both daily (P = .048) and intermittent montelukast (P = .028) compared with placebo. However, because of prespecified rules for multiplicity adjustments (requiring a positive primary endpoint), statistical significance for secondary endpoints cannot be concluded. All treatments were well tolerated. CONCLUSIONS: Montelukast did not reduce the number of asthma episodes culminating in an asthma attack over 1 year in children 6 months to 5 years of age, although numerical improvements occurred in some endpoints. PMID- 21624753 TI - Alteration of brain activation patterns in nonallergic rhinitis patients using functional magnetic resonance imaging before and after treatment with intranasal azelastine. AB - BACKGROUND: Although nonallergic rhinitis (NAR) patients tend to be more sensitive to chemical/olfactory stimuli, a suprathreshold olfactory response or the presence of specific olfactory receptor genes do not explain why their symptoms are triggered by such exposures. OBJECTIVE: To investigate differential neurogenic responses to azelastine in NAR patients, using functional magnetic resonance imaging (fMRI) in response to specific olfactory triggers. METHODS: A longitudinal study design on 12 subjects with a physician diagnosis of NAR previously demonstrated to be clinically responsive to intranasal azelastine (Astelin) was performed. Subjects underwent fMRI during exposure to unpleasant (hickory smoke) and pleasant (vanilla) odorants while off and then on azelastine for 2 weeks. The olfactory fMRI paradigm consisted of a visually triggered sniff every 21 seconds with synchronized delivery of a 4 second pulse of odorant. Each odorant was presented 18 times over 4-6-minute fMRI runs. Continuous fresh air was presented to wash out each odorant after presentation. RESULTS: Nonallergic rhinitis patients exhibited increased blood flow to several regions of the brain in response to both pleasant and unpleasant odorants, specifically in odor sensitive regions, while off intranasal azelastine. Treatment with intranasal azelastine significantly attenuated blood flow to regions of the brain relevant to either olfactory sensation or sensory processing in response to these odorants compared with fresh air. CONCLUSION: The general reduction compared with increase in brain activation in NAR patients on versus off azelastine suggests that a possible effect of this medication may be reduction of brain responses to odorants. PMID- 21624754 TI - Systemic reactions associated with subcutaneous allergen immunotherapy: timing and risk assessment. AB - BACKGROUND: Subcutaneous allergen immunotherapy (SCIT) is associated with risk of systemic reaction. Although risk factors have been identified, the incidence of immunotherapy-related systemic reactions has not changed in recent years. OBJECTIVES: To examine patterns of systemic reaction and determine whether risk of systemic reaction from SCIT is associated with patterns of response to skin tests to inhalant allergens recorded before receiving SCIT. METHODS: We carried out a retrospective review from January 2001 to December 2007. Patterns of systemic reaction from immunotherapy were examined. Cases were matched with controls by age (+/-10 years), sex, and time of injection (+/-1 week) to determine whether a pattern of more than 33% 3+ and 4+ skin test responses is associated with elevated risk for systemic reaction. RESULTS: Rate of systemic reaction from SCIT was 0.28% (46/16,375) per injection visit. Twenty patients had 46 systemic reactions. All severe reactions occurred within 30 minutes. The estimated odds of systemic reaction were almost 6 times higher for patients with more than 33% 3 to 4+ positive skin tests (OR = 5.83; 95%CI: 1.23-27.59, P = .026). For each additional 4+ skin test, the estimated odds for systemic reaction increased by 17% (P = .020). CONCLUSIONS: A small number of patients receiving SCIT account for a large proportion of systemic reactions. Skin test patterns demonstrating a greater number of larger skin tests responses to inhalant skin testing are associated with significantly elevated risk for systemic reaction in patients receiving SCIT. PMID- 21624755 TI - Fatal gas gangrene related to self-injection treatment of anaphylaxis. PMID- 21624756 TI - Anaphylaxis to Mopane worms (Imbrasia belina). PMID- 21624757 TI - Does the EASI score reflect itch severity? PMID- 21624758 TI - Successful desensitization with temozolomide. PMID- 21624759 TI - A novel method of desensitization for fluconazole hypersensitivity in a patient with AIDS. PMID- 21624760 TI - Identification of mold on seasonal indoor coniferous trees. PMID- 21624761 TI - Enterovirus-specific IgG in intravenous immunoglobulin preparations. PMID- 21624762 TI - Glyceraldehyde-3-phosphate dehydrogenase as a major allergen in rambutan-induced anaphylaxis. PMID- 21624763 TI - Adalimumab desensitization after anaphylactic reaction. PMID- 21624764 TI - The American Board of Allergy and Immunology maintenance of certification program. PMID- 21624765 TI - Allostery in the lac operon: population selection or induced dissociation? AB - Allostery, the modulation of function of a protein at one site by the binding of a ligand at a different site, is a property of many proteins. Two kinetically distinct models have been proposed: i) The induced fit model in which the ligand binds to the protein and then induces the conformational change. ii) The population selection model, in which the protein spontaneously undergoes a conformational change, which is then 'captured' by the ligand. Using measured kinetic constants for the lac repressor the contribution of population selection vs. induced dissociation is quantified by simulating the kinetics of allostery. At very low inducer concentration, both mechanisms contribute significantly. Total induction, though, is small under these conditions. At increasing levels of induction the induced dissociation mechanism soon dominates, first due to binding of one inducer, and then from two inducers binding. PMID- 21624766 TI - Needs-based sewerage prioritization: alternative to conventional cost-benefit analysis. AB - This paper presents an empirical approach to select and prioritize sewerage projects within set budgetary limitations. The methodology includes a model which quantifies benefits of a sewerage project as an index or dimensionless number. The index considers need and urgency of sewerage and other project goals. Benefit is defined as the difference in anticipated impact between the current condition (without the project) and the expected condition with the project. Anticipated benefits primarily include reduction in environmental pollution, reduction of human diseases and morbidity, and other tangible and intangible improvement. This approach is a powerful decision tool for sewerage prioritization and an effective alternative to conventional cost-benefit analysis. Unlike conventional analysis, this approach makes no attempt to convert project benefits and other impacts into a monetary measure. This work recognizes that the decision to provide sewerage based solely on net benefits is not practical. Instead, benefit-cost ratios (B/C) are calculated utilizing cost-effectiveness approach. Using these ratios, 16 unserviced areas of Ensenada, Mexico are ranked. The prioritization rankings produced by this method must be further scrutinized and carefully reviewed for logic, accuracy of input data, and practicality of implementation. A similar framework may also be useful for prioritizing other public works projects. PMID- 21624767 TI - Suppression of SOCS3 in macrophages prevents cancer metastasis by modifying macrophage phase and MCP2/CCL8 induction. AB - Inflammation has been demonstrated to play important roles in tumorigenesis, tumor progression, and metastasis. STAT3 has been shown to be frequently activated in a variety of human cancer cells and STAT3 signaling promotes the growth and survival of tumor cells. However, the role of STAT3 of myeloid cells associated with tumors is currently unknown. Suppressor of cytokine signaling-3 (SOCS3) has been shown to be a negative regulator of STAT3. In this study, we used macrophage specific SOCS3 conditional knockout (cKO) mice to investigate the effect of the hyperactivation of STAT3 in macrophages on tumor development and metastasis. In a subcutaneous transplantation model of B16F10 melanoma cells, although tumor sizes were not significantly different, SOCS3-cKO mice survived longer than wild-type (WT) mice did. SOCS3-cKO mice exhibited fewer lung and liver metastatic tumor nodules than WT mice when B16F10 was challenged intravenously. SOCS3(-/-) macrophages stimulated with tumor lysates in vitro exhibited prolonged STAT3 phosphorylation and produced less amount of TNFalpha and IL-6, and higher amount of MCP2/CCL8 than WT macrophages. MCP/CCL8 was induced via STAT3 and exhibited anti-tumor metastatic effect in WT mice. These data suggest that hyperactivation of STAT3 in myeloid cells simultaneously exerted an anti-inflammatory as well as anti-tumor effects. Thus, the targeted inhibition of SOCS3 activity in macrophages may be therapeutic for the suppression of tumor metastasis. PMID- 21624768 TI - Stromal cells do not share KRAS mutations that the epithelial component harbors in colorectal carcinomas. AB - Epithelia and stroma are the main components of carcinomas that have impact on carcinogenesis. Many of the genetic changes harboring in the epithelia may possibly be seen in stromal cells during neoplastic transformation. We intended to investigate weather KRAS mutations are shared by the stromal cells in colorectal adenocarcinomas. Forty cases with KRAS mutation were studied. Glandular/epithelial and the stromal components of each primary tumor were collected and KRAS mutation analysis was performed. None of the cases revealed KRAS mutations in stromal integral. We concluded that stromal cells in colorectal carcinoma do not share KRAS mutations that the epithelial component harbors. PMID- 21624769 TI - Mutual enhancement between heparanase and vascular endothelial growth factor: a novel mechanism for melanoma progression. AB - Heparanase is closely related to growth factors in the role of promoting tumor progression. Among them, vascular endothelial growth factor (VEGF) is necessary for tumor vascularity and metastasis. Release of VEGF by heparanase can initiate relative signaling pathways, resulting in an up-regulation of transcriptional factors related with heparanase. Therefore, VEGF likely has a potential function as a regulator of heparanase expression in melanoma. We hypothesized that a novel mechanism exists where heparanase and VEGF are mutually enhanced in melanoma. Our study was conducted to validate the hypothetical mutual enhancement and elucidate its effect on melanoma progression. We found that the addition of exogenous VEGF and its cDNA transfection induce heparanase over-expression by means of western blot and real-time RT-PCR, while anti-VEGF siRNA reduces heparanase expression in A2058 and WM793 melanoma cell lines. Likewise, VEGF expression is also regulated by heparanase in these two cell lines. Additionally, the cells with mutual enhancement phenotypes exhibit higher proliferation and transmigration capacity. PD98059, a specific inhibitor of the MEK/ERK signaling pathway, is involved in this mutual enhancement. These data are the first to show that heparanase and VEGF have a mutual enhancement in melanoma cells, which may be a novel mechanism for promoting melanoma progression. PMID- 21624770 TI - Reducing cancer risk in homosexual men. PMID- 21624771 TI - Optimum chemotherapy regimen for early breast cancer. PMID- 21624773 TI - Tuberculosis and cancer: a complex and dangerous liaison. PMID- 21624774 TI - The effects of total and REM sleep deprivation on laser-evoked potential threshold and pain perception. AB - We investigated the effects of total and rapid eye movement (REM) sleep deprivation on the thermal nociceptive threshold and pain perception using the objective laser-evoked potential (LEP) and the subjective visual analogue scale (VAS). Twenty-eight male adult volunteers were assigned into Control (CTRL), Total (T-SD), and REM (REM-SD) Sleep Deprivation groups. The T-SD and REM-SD volunteers were totally or selectively deprived of sleep for 2 and 4 consecutive nights, respectively. Pain parameters were measured daily during the experimental period. Volunteers were stimulated on the back of the hand by blocks of 50 diode laser pulses. Intensities increased between successive blocks, ranging from nonnoxious to noxious levels, and the LEP threshold was identified based on the evoked-response onset. Both the LEP threshold and VAS ratings were significantly increased after the second night of T-SD. No significant variations were observed in the REM-SD group, suggesting a predominant role for slow wave sleep rather than selective REM-SD in pain perception. Also, for both sleep-deprived groups, the mean values of the LEP threshold and VAS ratings showed a gradual increase that was proportional to the SD deprivation time, followed by a decrease after 1 night of sleep restoration. These findings demonstrate a hyperalgesic modification to pain perception (as reflected by the augmented VAS) and a concomitant increase in the LEP threshold following T-SD, an apparently contradictory effect that can be explained by differences in the ways that attention affects these pain measurements. PMID- 21624775 TI - The DrugCheck Problem List: a new screen for substance use disorders in people with psychosis. AB - Despite considerable recent interest in the issue of comorbid substance use disorders in people with serious mental illness, there remains a need to refine approaches to screening. This paper describes the development and testing of a new screen for substance-related comorbidity, the 12-item DrugCheck Problem List (PL). Exploratory factor analysis with inpatient samples suggested a single factor structure, although confirmatory factor analysis in a further sample found similar fit from a two-factor model. Sensitivity and specificity in detecting DSM IV substance use disorders were both high and comparable to performances of the Severity of Dependence Scale and Alcohol Use Disorders Identification Test (Australian version). The list of problem areas provided by the PL has utility in driving further assessment and treatment planning, and offers suggested foci for motivational interviewing. While further testing is indicated, these data provide strong initial support for its use. PMID- 21624776 TI - [Lung transplantation in children: challenges and realities]. PMID- 21624777 TI - How backyard poultry flocks influence the effort required to curtail avian influenza epidemics in commercial poultry flocks. AB - This paper summarizes the evidence that the contribution of backyard poultry flocks to the on-going transmission dynamics of an avian influenza epidemic in commercial flocks is modest at best. Nevertheless, while disease control strategies need not involve the backyard flocks, an analysis of the contribution of each element of the next generation matrix to the basic reproduction number indicates that models which ignores the contribution of backyard flocks in estimating the effort required of strategies focused one host type (e.g. commercial flocks only) necessarily underestimate the level of effort to an extent that may matter to policy makers. PMID- 21624778 TI - Genetic tracking of the raccoon variant of rabies virus in eastern North America. AB - To gain insight into the incursion of the raccoon variant of rabies into the raccoon population in three Canadian provinces, a collection of 192 isolates of the raccoon rabies virus (RRV) strain was acquired from across its North American range and was genetically characterized. A 516-nucleotide segment of the non coding region between the G and L protein open reading frames, corresponding to the most variable region of the rabies virus genome, was sequenced. This analysis identified 119 different sequences, and phylogenetic analysis of the dataset supports the documented history of RRV spread. Three distinct geographically restricted RRV lineages were identified. Lineage 1 was found in Florida, Alabama and Georgia and appears to form the ancestral lineage of the raccoon variant of rabies. Lineage 2, represented by just two isolates, was found only in Florida, while the third lineage appears broadly distributed throughout the rest of the eastern United States and eastern Canada. In New York State, two distinct spatially segregated variants were identified; the one occupying the western and northern portions of the state was responsible for an incursion of raccoon rabies into the Canadian province of Ontario. Isolates from New Brunswick and Quebec form distinct, separate clusters, consistent with their independent origins from neighboring areas of the United States. The data are consistent with localized northward incursion into these three separate areas with no evidence of east-west viral movement between the three Canadian provinces. PMID- 21624779 TI - Declining HIV prevalence and incidence in perinatal women in Harare, Zimbabwe. AB - BACKGROUND: In several recent papers it has been suggested that HIV prevalence and incidence are declining in Zimbabwe as a result of changing sexual behavior. We provide further support for these suggestions, based on an analysis of more extensive, age-stratified, HIV prevalence data from 1990 to 2009 for perinatal women in Harare, as well as data on incidence and mortality. METHODOLOGY/PRINCIPAL FINDINGS: Pooled prevalence, incidence and mortality were fitted using a simple susceptible-infected (SI) model of HIV transmission; age stratified prevalence data were fitted using double-logistic functions. We estimate that incidence peaked at 5.5% per year in 1991 declining to 1% per year in 2010. Prevalence peaked in 1998/9 [35.9% (CI95: 31.3-40.7)] and decreased by 67% to 11.9% (CI95: 10.1-13.8) in 2009. For women <20y, 20-24y, 25-29y, 30-34y and >=35y, prevalence peaked at 25.4%, 34.2%, 47.1%, 44.0% and 33.5% in 1993, 1996, 1997, 1998 and 1999, respectively, declining thereafter in every age group. Among women <25y, prevalence peaked in 1994 at 28.8% declining thereafter by 69% to 8.9% (CI95: 6.8-11.5) in 2009. CONCLUSION/SIGNIFICANCE: HIV prevalence declined substantially among perinatal women in Harare after 1998 consequent upon a decline in incidence starting in the early 1990s. Our model suggests that this was primarily a result of changes in behavior which we attribute to a general increase in awareness of the dangers of AIDS and the ever more apparent increases in mortality. PMID- 21624780 TI - An individual-based simulation of pneumonic plague transmission following an outbreak and the significance of intervention compliance. AB - The existence of primary pneumonic plague outbreaks raises concerns over the use of the causative bacteria as an aerosol-based bioweapon. We employed an individual-based model, parameterised using published personal contact information, to assess the severity of a deliberate release in a discrete community, under the influence of two proposed intervention strategies. We observed that the severity of the resulting epidemic is determined by the degree of personal compliance with said strategies, implying that prior preparedness activities are essential in order that public awareness and willingness to seek treatment is achieved quickly. PMID- 21624781 TI - The impact of school holidays on the social mixing patterns of school children. AB - School holidays are recognised to be of great epidemiological importance for a wide range of infectious diseases; this is postulated to be because the social mixing patterns of school children - a key population group - change significantly during the holiday period. However, there is little direct quantitative evidence to confirm this belief. Here, we present the results of a prospective survey designed to provide a detailed comparison of social mixing patterns of school children during school terms and during the school holidays. Paired data were collected, with participants recording their social contacts once during term time and once during the holiday period. We found that the daily number of recorded encounters approximately halved during the holidays, and that the number of close contact encounters fell by approximately one third. The holiday period also saw a change in the age structure of children's social contacts, with far fewer contacts of their own age, but an increase in the number of encounters with adults, particularly older adults. A greater amount of mixing between children at different schools was recorded during the holiday. We suggest, therefore, that whilst infections may spread rapidly within schools during term time, in the holiday period there are increased opportunities for transmission to other schools and other age groups. PMID- 21624782 TI - The distribution of Plasmodium falciparum infection durations. AB - OBJECTIVES: The duration of untreated Plasmodium falciparum infections in naturally exposed human populations is of interest for rational planning of malaria control interventions as it is related to the duration of infectivity. The extent of variability in duration is relevant where transmission is seasonal, and for the planning of elimination efforts. Methods for measuring these quantities from genotyping data have been restricted to exponential models of infection survival, as implied by constant clearance rates. Such models have greatly improved the understanding of infection dynamics on a population level but likely misrepresent the within-host dynamics of many pathogens. Conversely, the statistical properties of the distribution of infection durations, and how these are affected by exposure, should contain information on within-host dynamics. METHODS AND RESULTS: We extended existing methods for the analysis of longitudinal genotyping data on P. falciparum infections. Our method simultaneously estimates force of infection, detectability, and the distribution of infection durations. Infection durations are modeled using parametric survival distributions. The method is validated using simulated data, and applied to data from a cohort study in Navrongo, Northern Ghana. Distribution estimates from exponential, Weibull, lognormal, and gamma models are compared with the distribution of durations in malariatherapy data. CONCLUSIONS: The Weibull model fitted the data best. It estimated a shorter mean duration than the exponential model, which gave the worst fit. The distribution estimates appeared positively skewed when compared with the distribution of durations in malariatherapy data, suggesting that a significant proportion of infections is cleared shortly after inoculation. We conclude that malariatherapy data, the most important source of information on P. falciparum within-host dynamics, may not be representative of the actual processes in natural populations, and should be used with care. Further, conclusions from transmission models assuming exponential infection survival may be biased. PMID- 21624783 TI - The epidemiological dynamics of infectious trachoma may facilitate elimination. AB - INTRODUCTION: Trachoma programs use mass distributions of oral azithromycin to treat the ocular strains of Chlamydia trachomatis that cause the disease. There is debate whether infection can be eradicated or only controlled. Mass antibiotic administrations clearly reduce the prevalence of chlamydia in endemic communities. However, perfect coverage is unattainable, and the World Health Organization's goal is to control infection to a level where resulting blindness is not a public health concern. Here, we use mathematical models to assess whether more ambitious goals such as local elimination or even global eradication are possible. METHODS: We fit a class of non-linear, stochastic, susceptible infectious-susceptible (SIS) models which allow positive or negative feedback, to data from a recent community-randomized trial in Ethiopia, and make predictions using model averaging. RESULTS: The models predict that reintroduced infection may not repopulate the community, or may do so sufficiently slowly that surveillance might be effective. The preferred model exhibits positive feedback, allowing a form of stochastic hysteresis in which infection returns slowly after mass treatment, if it returns at all. Results for regions of different endemicity suggest that elimination may be more feasible than earlier models had predicted. DISCUSSION: If trachoma can be eradicated with repeated mass antibiotic distributions, it would encourage similar strategies against other bacterial diseases whose only host is humans and for which effective vaccines are not available. PMID- 21624784 TI - Different transmission patterns in the early stages of the influenza A(H1N1)v pandemic: a comparative analysis of 12 European countries. AB - Following the emergence of a novel strain of influenza A(H1N1) in Mexico and the United States in April 2009, its epidemiology in Europe during the summer was limited to sporadic and localised outbreaks. Only the United Kingdom experienced widespread transmission declining with school holidays in late July. Using statistical modelling where applicable we explored the following causes that could explain this surprising difference in transmission dynamics: extinction by chance, differences in the susceptibility profile, age distribution of the imported cases, differences in contact patterns, mitigation strategies, school holidays and weather patterns. No single factor was able to explain the differences sufficiently. Hence an additive mixed model was used to model the country-specific weekly estimates of the effective reproductive number using the extinction probability, school holidays and weather patterns as explanatory variables. The average extinction probability, its trend and the trend in absolute humidity were found to be significantly negatively correlated with the effective reproduction number - although they could only explain about 3% of the variability in the model. By comparing the initial epidemiology of influenza A (H1N1) across different European countries, our analysis was able to uncover a possible role for the timing of importations (extinction probability), mixing patterns and the absolute humidity as underlying factors. However, much uncertainty remains. With better information on the role of these epidemiological factors, the control of influenza could be improved. PMID- 21624785 TI - Acute ethanol exposure prevents PMA-mediated augmentation of N-methyl-D-aspartate receptor function in primary cultured cerebellar granule cells. AB - Many intracellular proteins and signaling cascades contribute to the ethanol sensitivity of native N-methyl-D-aspartate receptors (NMDARs). One putative protein is the serine/threonine kinase, protein kinase C (PKC). The purpose of this study was to assess if PKC modulates the ethanol sensitivity of native NMDARs expressed in primary cultured cerebellar granule cells (CGCs). With the whole-cell patch-clamp technique, we assessed if ethanol inhibition of NMDA induced currents (I(NMDA)) (100 MUM NMDA plus 10 MUM glycine) were altered in CGCs in which the novel and classical PKC isoforms were activated by phorbol-12 myristate-13-acetate (PMA). Percent inhibition by 10, 50, or 100 mM ethanol of NMDA-induced steady-state current amplitudes (I(SS)) or peak current amplitudes (I(Pk)) of NMDARs expressed in CGCs in which PKC was activated by a 12.5 min, 100 nM PMA exposure at 37 degrees C did not differ from currents obtained from receptors contained in control cells. However, PMA-mediated augmentation of I(Pk) in the absence of ethanol was abolished after brief applications of 10 or 1 mM ethanol coapplied with agonists, and this suppression of enhanced receptor function was observed for up to 8 min post-ethanol exposure. Because we had previously shown that PMA-mediated augmentation of I(NMDA) of NMDARs expressed in these cells is by activation of PKCalpha, we assessed the effect of ethanol (1, 10, 50, and 100 mM) on PKCalpha activity. Ethanol decreased PKCalpha activity by 18% for 1 mM ethanol and activity decreased with increasing ethanol concentrations with a 50% inhibition observed with 100 mM ethanol. The data suggest that ethanol disruption of PMA-mediated augmentation of I(NMDA) may be due to a decrease in PKCalpha activity by ethanol. However, given the incomplete blockade of PKCalpha activity and the low concentration of ethanol at which this phenomenon is observed, other ethanol-sensitive signaling cascades must also be involved. PMID- 21624786 TI - Lymphocyte measures in treatment-naive 13-15-year old adolescents with alcohol use disorders. AB - Many adolescents have chronic exposure to hazardous levels of alcohol. This is likely to be a significant predictor of health outcomes, including those related to immunity. We assessed substance use and biochemical immunological parameters in heavy drinking adolescents (meeting DSM-IV criteria for alcohol dependence) and light/nondrinking control adolescents in Cape Town. Lifetime alcohol dose, measured in standard units of alcohol, was orders of magnitude higher in alcohol dependent (AD) participants than controls. All adolescent AD had a "weekends only" style of alcohol consumption. The AD group was chosen to represent relatively "pure" AD, with minimal other drug use and no psychiatric diagnoses. With these narrow parameters in place, we found that AD adolescents were lymphopenic compared with controls, with significantly lower mean numbers of absolute circulating CD3+, CD4+, and CD8+ T-lymphocytes. On conclusion, we found that adolescent AD individuals with excessive alcohol intake, in a weekend binge drinking style but without comorbid drug or psychiatric disorders, may be at increased risk of lymphopenia. This alcohol misuse may increase infectious disease susceptibility (including TB and HIV) by reducing immune system capabilities. Complex interactions of alcohol with other documented high-risk activities may further compound health risks. PMID- 21624787 TI - Assessment of pulmonary hypertension during exercise: ready for clinical prime time? PMID- 21624788 TI - Thrombus in normal coronary arteries: retrospective study and review of case reports. AB - BACKGROUND: Myocardial infarction is rarely caused by non-occlusive thrombus in angiographically normal coronary arteries. The cases reported in the literature are scarce and follow-up was usually short. The efficacy and tolerability of the exclusively medical treatment strategy used in most cases remain unknown. AIMS: To evaluate efficacy of medical treatment and long-term prognosis in these patients. METHODS: We retrospectively selected and analysed patients hospitalized in our centre between 1998 and 2008 for myocardial infarction caused by non occlusive thrombus in angiographically normal coronary arteries (defined as stenosis<30%), who were exclusively medically treated. A long-term follow-up was performed. A review of the literature regarding such cases was carried out. RESULTS: Sixteen patients were identified; apart from smoking, they had few conventional cardiovascular risk factors. Two patients died in hospital. The 14 survivors were followed up for an average of 4.9 years and only one death (non cardiac cause) and one stroke (related to supraventricular arrhythmia) occurred in this period. Medical treatment included the use of glycoprotein IIb/IIIa inhibitors in 75% of cases. The literature review revealed 36 similar cases due to multiple aetiologies-particularly coronary artery spasm and prothrombotic coagulopathies. CONCLUSION: Patients with myocardial infarction secondary to non occlusive thrombus in angiographically normal coronary arteries seem to have a good long-term prognosis after the acute phase when treated with an exclusively medical strategy. However, initial clinical presentation was often severe, leading to early in-hospital death. PMID- 21624789 TI - Inflammation abnormalities and inducibility of atrial fibrillation after epicardial ganglionated plexi ablation. AB - BACKGROUND: Epicardial ganglionated plexi (GP) ablation can prevent atrial fibrillation inducibility. However, the long-term effects of GP ablation on atrial fibrillation have not been elucidated. METHODS: Thirteen adult dogs of either sex, weighing 13-17kg, were randomly assigned to a sham-operated group (n=6) or a GP ablation group (n=7). After right thoracotomy, the atrial effective refractory period (AERP) was measured and atrial fibrillation was induced by right atrial rapid burst pacing. Atrial fibrillation and AERP were remeasured after anterior right and inferior right GP ablation in the GP ablation group. The animals were allowed to recover for 8 weeks, after which atrial fibrillation and AERP were measured again. Concentrations of C-reactive protein, tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 were measured in the blood and atrial tissues. RESULTS: After 8 weeks, atrial fibrillation was induced in all animals in the GP ablation group. AERP and dispersion of AERP (dAERP; maximum AERP minus minimum AERP) were increased after GP ablation but AERP recovered after 8 weeks. There were no significant differences in the concentrations of C-reactive protein, TNF-alpha or interleukin-6 in venous blood between the two groups and the concentration of C-reactive protein in the atrium did not change before and after GP ablation. However, the concentrations of TNF-alpha and interleukin-6 in the atrium increased significantly 8 weeks after GP ablation (P<0.05). CONCLUSION: Increased concentrations of TNF-alpha and interleukin-6 in the atrium after GP ablation provide a new causative factor in terms of atrial fibrillation vulnerability. PMID- 21624790 TI - Impact of bifurcation lesions on angiographic characteristics and procedural success in primary percutaneous coronary intervention for ST-segment elevation myocardial infarction. AB - BACKGROUND: Bifurcation lesions (BFLs) remain a challenging lesion subset, often associated with lower success rates than less complex lesions. There are few data regarding the impact of BFLs in the setting of ST-segment elevation myocardial infarction (STEMI). AIMS: To assess the impact of BFLs on angiographic characteristics and procedural success in primary percutaneous coronary interventions (PCIs). METHODS: Out of 1070 primary PCIs performed between November 2006 and December 2008, 114 patients (10.7%) with a BFL (side branch >=2.0mm) were identified and matched with 114 patients without a BFL, according to age, sex and infarct-related artery. RESULTS: Baseline characteristics were similar in both groups. Using the Medina classification, true BFLs ([1,1,1]; [1,0,1]; [0,1,1]) were found in 46.5% of cases. Mean contrast volume (265+/-91 and 207+/-68mL), procedural time (51.0+/-26.6 vs 35.3+/-11.5min) and fluoroscopy time (16.2+/-11.2 vs 9.8+/-5.1min) were significantly higher in the BFL group than the non-BFL group (p<0.0001). However, time to reperfusion and angiographic success rates (residual stenosis <= 30% and Thrombolysis in Myocardial Infarction flow grade 3 in main branch) were similar in BFL and non-BFL patients (13.7+/-7.9 vs 12.1+/-5.7min, respectively, p=0.087; 96.5 vs 99.1%, respectively, p=0.18), with no periprocedural events (in-hospital death, emergent coronary artery bypass graft or repeat PCI<24h). CONCLUSION: Despite being challenging lesions, BFLs in STEMI were associated with similar time to reperfusion and procedural success but led to significantly greater contrast use and prolonged procedural time compared with non-BFLs. PMID- 21624791 TI - Patent foramen ovale and obstructive sleep apnoea: from pathophysiology to diagnosis of a potentially dangerous association. AB - Patent foramen ovale and obstructive sleep apnoea are frequently encountered in the general population. Owing to their prevalence, they may coexist fortuitously; however, the prevalence of patent foramen ovale seems to be higher in patients with obstructive sleep apnoea. We have reviewed the epidemiological data, pathophysiology, and the diagnostic and therapeutic options for both patent foramen ovale and obstructive sleep apnoea. We focus on the interesting pathophysiological links that could explain a potential association between both pathologies and their implications, especially on the risk of stroke. PMID- 21624792 TI - An unusual case of hypertensive cardiomyopathy. PMID- 21624793 TI - Endovascular treatment of superior vena cava obstruction helping permanent pacing. PMID- 21624794 TI - Platypnoea-orthodeoxia syndrome due to a patent foramen oval with marked lipomatous hypertrophy. PMID- 21624795 TI - Value of cardiac MRI in peripartum cardiomyopathy. PMID- 21624797 TI - The value of connections. PMID- 21624798 TI - Notch and basal cells take center stage during airway epithelial regeneration. AB - Airway epithelium is maintained by basal cells, but the mechanisms responsible for repair are poorly characterized. In this issue of Cell Stem Cell, Rock et al. (2011) show that Notch signals regulate differentiation, but not self-renewal of basal cells after injury, and that this role appears conserved in mouse and human lung. PMID- 21624799 TI - Small RNAs loom large during reprogramming. AB - In two independent Cell Stem Cell reports, the Morrisey and Mori groups show that human and mouse somatic cells can be reprogrammed to produce induced pluripotent stem cells by expressing microRNAs, completely eliminating the need for ectopic protein expression (Anokye-Danso et al., 2011; Miyoshi et al., 2011). PMID- 21624800 TI - Taking HSCs down a Notch in leukemia. AB - The Notch signaling pathway is activated in the majority of T cell acute lymphoblastic leukemias (T-ALL). Adding to the complexity of Notch signaling in hematopoiesis, recently in Nature, Klinakis et al. (2011) demonstrate a tumor suppressor function for the Notch pathway in myeloid malignancy. PMID- 21624801 TI - Epigenetic modifications in distinction: histone versus DNA methylation in ESCs. AB - In this issue of Cell Stem Cell, Karimi et al. (2011) show that DNA methylation and histone H3 lysine 9 trimethylation (H3K9me3) have distinct genomic targets in mouse ESCs. In particular, loss of H3K9me3 leads to derepression of select endogenous retroviruses and subsequent ectopic transcription of adjacent genes. PMID- 21624802 TI - The nomenclature system should be sustainable, but also practical. PMID- 21624803 TI - Reaching for consensus on a naming convention for pluripotent cells. PMID- 21624804 TI - Hematopoietic stem cell transplantation and implications for cell therapy reimbursement. AB - As costly stem cell treatments progress from experimental concepts toward licensed products and routine procedures, governmental and private payers grapple with shrinking budgets to cover more lives. We describe efforts underway in the US to create mechanisms for reimbursement of cell therapies and discuss other reimbursement-related issues for the stem cell community. PMID- 21624805 TI - Researching the relationships between tissue providers, clinicians, and stem cell scientists. AB - Access to human reproductive tissue is essential for many forms of stem cell research. We identify questions for future studies of tissue providers, procurers, and end-user scientists, and suggest that international comparative studies of all three parties, and of the relationships between them, will improve the ethical supply of tissue. PMID- 21624806 TI - Assessing the safety of stem cell therapeutics. AB - Unprecedented developments in stem cell research herald a new era of hope and expectation for novel therapies. However, they also present a major challenge for regulators since safety assessment criteria, designed for conventional agents, are largely inappropriate for cell-based therapies. This article aims to set out the safety issues pertaining to novel stem cell-derived treatments, to identify knowledge gaps that require further research, and to suggest a roadmap for developing safety assessment criteria. It is essential that regulators, pharmaceutical providers, and safety scientists work together to frame new safety guidelines, based on "acceptable risk," so that patients are adequately protected but the safety "bar" is not set so high that exciting new treatments are lost. PMID- 21624807 TI - Reflections of an ISSCR president, 2010-2011. PMID- 21624809 TI - Notch-dependent differentiation of adult airway basal stem cells. AB - The epithelium lining the airways of the adult human lung is composed of ciliated and secretory cells together with undifferentiated basal cells (BCs). The composition and organization of this epithelium is severely disrupted in many respiratory diseases. However, little is known about the mechanisms controlling airway homeostasis and repair after epithelial damage. Here, we exploit the mouse tracheobronchial epithelium, in which BCs function as resident stem cells, as a genetically tractable model of human small airways. Using a reporter allele we show that the low level of Notch signaling at steady state is greatly enhanced during repair and the generation of luminal progenitors. Loss-of-function experiments show that Notch signaling is required for the differentiation, but not self-renewal, of BCs. Moreover, sustained Notch activation in BCs promotes their luminal differentiation, primarily toward secretory lineages. We also provide evidence that this function of Notch signaling is conserved in BCs from human airways. PMID- 21624810 TI - Functional crosstalk between Bmi1 and MLL/Hoxa9 axis in establishment of normal hematopoietic and leukemic stem cells. AB - Bmi1 is required for efficient self-renewal of hematopoietic stem cells (HSCs) and leukemic stem cells (LSCs). In this study, we investigated whether leukemia associated fusion proteins, which differ in their ability to activate Hox expression, could initiate leukemia in the absence of Bmi1. AML1-ETO and PLZF RARalpha, which do not activate Hox, triggered senescence in Bmi1(-/-) cells. In contrast, MLL-AF9, which drives expression of Hoxa7 and Hoxa9, readily transformed Bmi1(-/-) cells. MLL-AF9 could not initiate leukemia in Bmi1(-/ )Hoxa9(-/-) mice, which have further compromised HSC functions. But either gene could restore the ability of MLL-AF9 to establish LSCs in the double null background. As reported for Bmi1, Hoxa9 regulates expression of p16(Ink4a)/p19(ARF) locus and could overcome senescence induced by AML1-ETO. Together, these results reveal an important functional interplay between MLL/Hox and Bmi1 in regulating cellular senescence for LSC development, suggesting that a synergistic targeting of both molecules is required to eradicate a broader spectrum of LSCs. PMID- 21624811 TI - Transcription factor-induced lineage selection of stem-cell-derived neural progenitor cells. AB - The generation of specific types of neurons from stem cells offers important opportunities in regenerative medicine. However, future applications and proper verification of cell identities will require stringent ways to generate homogeneous neuronal cultures. Here we show that transcription factors like Lmx1a, Phox2b, Nkx2.2, and Olig2 can induce desired neuronal lineages from most expressing neural progenitor cells by a mechanism resembling developmental binary cell-fate switching. Such efficient selection of cell fate resulted in remarkable cellular enrichment that enabled global gene-expression validation of generated neurons and identification of previously unrecognized features in the studied cell lineages. Several sources of stem cells have a limited competence to differentiate into specific neuronal cell types; e.g., dopamine neurons. However, we show that the combination of factors that normally promote either regional or dedicated neuronal specification can overcome limitations in cellular competence and also promote efficient reprogramming in more remote neural contexts, including human neural progenitor cells. PMID- 21624812 TI - DNA methylation and SETDB1/H3K9me3 regulate predominantly distinct sets of genes, retroelements, and chimeric transcripts in mESCs. AB - DNA methylation and histone H3 lysine 9 trimethylation (H3K9me3) play important roles in silencing of genes and retroelements. However, a comprehensive comparison of genes and repetitive elements repressed by these pathways has not been reported. Here we show that in mouse embryonic stem cells (mESCs), the genes upregulated after deletion of the H3K9 methyltransferase Setdb1 are distinct from those derepressed in mESC deficient in the DNA methyltransferases Dnmt1, Dnmt3a, and Dnmt3b, with the exception of a small number of primarily germline-specific genes. Numerous endogenous retroviruses (ERVs) lose H3K9me3 and are concomitantly derepressed exclusively in SETDB1 knockout mESCs. Strikingly, ~15% of upregulated genes are induced in association with derepression of promoter-proximal ERVs, half in the context of "chimeric" transcripts that initiate within these retroelements and splice to genic exons. Thus, SETDB1 plays a previously unappreciated yet critical role in inhibiting aberrant gene transcription by suppressing the expression of proximal ERVs. PMID- 21624813 TI - miR-371-3 expression predicts neural differentiation propensity in human pluripotent stem cells. AB - The use of pluripotent stem cells in regenerative medicine and disease modeling is complicated by the variation in differentiation properties between lines. In this study, we characterized 13 human embryonic stem cell (hESC) and 26 human induced pluripotent stem cell (hiPSC) lines to identify markers that predict neural differentiation behavior. At a general level, markers previously known to distinguish mouse ESCs from epiblast stem cells (EPI-SCs) correlated with neural differentiation behavior. More specifically, quantitative analysis of miR-371-3 expression prospectively identified hESC and hiPSC lines with differential neurogenic differentiation propensity and in vivo dopamine neuron engraftment potential. Transient KLF4 transduction increased miR-371-3 expression and altered neurogenic behavior and pluripotency marker expression. Conversely, suppression of miR-371-3 expression in KLF4-transduced cells rescued neural differentiation propensity. miR-371-3 expression level therefore appears to have both a predictive and a functional role in determining human pluripotent stem cell neurogenic differentiation behavior. PMID- 21624814 TI - Trajectories of approaching death in the emergency department: clinician narratives of patient transitions to the end of life. AB - CONTEXT: Transitions near the end of life have not been well articulated and the end-of-life (EOL) phase is not well understood in the emergency department (ED). The sudden and unforeseen is common in the ED. OBJECTIVES: The purpose of this qualitative research project is to identify different trajectories of approaching death in an effort to describe the EOL experience in the ED. METHODS: An interpretive phenomenological approach was used to assess the results of interviews with and observations of ED staff who were intimately involved in caring for patients approaching death and dying. RESULTS: Seven trajectories of approaching death in the ED emerged from the data: 1) dead on arrival; 2) prehospital resuscitation with subsequent ED death; 3) prehospital resuscitation with survival until admission; 4) terminally ill and comes to the ED; 5) frail and hovering near death; 6) alive and interactive on arrival, but arrests in the ED; and 7) potentially preventable death by omission or commission. CONCLUSION: A descriptive articulation of the various trajectories will help clinicians be more astute in their recognition of the clinical situation and react appropriately, will help identify the transitions to the EOL phase, and will help to explore the possibilities open to the patient, family, and clinicians. In addition, understanding the trajectories and discussion of the clinicians' actions and communication strategies can elucidate which of the trajectories could benefit from anticipatory planning. PMID- 21624815 TI - Analysis of speech fluency in Williams syndrome. AB - Williams syndrome (WS) is a neurodevelopmental genetic disorder, often referred as being characterized by dissociation between verbal and non-verbal abilities, although the number of studies disputing this proposal is emerging. Indeed, although they have been traditionally reported as displaying increased speech fluency, this topic has not been fully addressed in research. In previous studies carried out with a small group of individuals with WS, we reported speech breakdowns during conversational and autobiographical narratives suggestive of language difficulties. In the current study, we characterized the speech fluency profile using an ecologically based measure--a narrative task (story generation) was collected from a group of individuals with WS (n = 30) and typically developing group (n = 39) matched in mental age. Oral narratives were elicited using a picture stimulus--the cookie theft picture from Boston Diagnosis Aphasia Test. All narratives were analyzed according to typology and frequency of fluency breakdowns (non-stuttered and stuttered disfluencies). Oral narratives in WS group differed from typically developing group, mainly due to a significant increase in the frequency of disfluencies, particularly in terms of hesitations, repetitions and pauses. This is the first evidence of disfluencies in WS using an ecologically based task (oral narrative task), suggesting that these speech disfluencies may represent a significant marker of language problems in WS. PMID- 21624816 TI - Delayed motor skill acquisition in kindergarten children with language impairment. AB - The acquisition and consolidation of a new grapho-motor symbol into long-term memory was studied in 5-year-old children with language impairment (LI) and peers matched for age and visual-motor integration skills. The children practiced the production of a new symbol and were tested 24h and two weeks post-practice day. Differences in performance speed emerged between the groups: children with LI showed a later onset of rapid learning in the practice phase, and only the comparison group exhibited delayed, consolidation, gains 24h post-training. At two weeks post-training, children with LI improved, closing the gap in performance speed. Speed-accuracy trade-off was characteristic of speed improvements in LI. These results indicate atypical and delayed acquisition in children with LI, and support the view that deficient skill acquisition in LI goes beyond the language system. PMID- 21624817 TI - Cross-national invariance of Attention-Deficit/Hyperactivity Disorder factors in Japanese and U.S. university students. AB - Prior research with children generally supports the two-dimensional structure of Attention-Deficit/Hyperactivity Disorder (ADHD; inattentive and hyperactive/impulsive factors) of the DSM-IV-TR as well as invariance of the two factor structure across nations and cultures. Research with adults supports either a two-factor or three-factor structure depending on reporting source and breadth of symptoms assessed. However, research with adults is limited and there are few studies addressing cross-national invariance in adults. The purposes of this study were to (1) assess relative fit of two- versus three-factor solutions for self-report of childhood and recent ADHD symptoms in adults; and (2) further establish cross-national invariance of factors. Participants included 271 U.S. and 712 Japanese university students who completed a rating scale assessing the 18 DSM-IV-TR ADHD symptoms. Confirmatory factor analysis using Mplus (Version 6) and the mean and variance-adjusted weighted least squares (WLSMV) procedure showed invariance of two- and three-factor models across U.S. and Japanese samples. The two- and three-factor models showed similar fit indices. Neither a two-factor or three-factor model was clearly superior. The two-factor model was favored, however, because it is more parsimonious and consistent with current theory, and because of high correlations between hyperactive and impulsive factors in the three-factor models. Invariance across nations is consistent with previous studies and supports ADHD as a universally valid syndrome rather than a cultural construct. These results add to the limited knowledge of assessment of ADHD symptoms in Japan. PMID- 21624818 TI - Sit-to-stand movement in children with cerebral palsy: a critical review. AB - Sit-to-stand (STS) movement is widely performed in daily life and an important pre requisite for acquisition of functional abilities. However, STS is a biomechanical demanding task which requires high levels of neuromuscular coordination, muscle strength and postural control. As children with cerebral palsy (CP) exhibit a series of impairments in body structures and functions, STS movement performance could be impaired in this population. Thus, this article aimed to review studies that had described how STS movement is performed by children with CP, the factors that influence it and the methodological procedures adopted in it analyses. A search was performed by one reviewer in relevant databases. In all, 12 articles were identified and 9 were selected for the present review. It was detected a large variation in sample characteristics and methodological issues among studies. In fact, standardization of the method applied to STS movement analysis is not fully established. With regard to STS performance, children with CP exhibited variations among them and also when compared with their typical peers. Moreover extrinsic factors appear to influence STS movement performance in these children and its manipulation could be incorporated into rehabilitation protocols. Moreover, the relationship between STS movement and functionality in reviewed articles was not reported. Therefore the review allowed to observe that STS movement has been under-explored in children with CP, with a lack of standardized methodologies and a not well established relationship between this movement and functionality. Thus, further studies about STS movement in CP are necessary. PMID- 21624819 TI - Classification of topographical pattern of spasticity in cerebral palsy: a registry perspective. AB - This study used data from a population-based cerebral palsy (CP) registry and systematic review to assess the amount of heterogeneity between registries in topographical patterns when dichotomised into unilateral (USCP) and bilateral spastic CP (BSCP), and whether the terms diplegia and quadriplegia provide useful additional epidemiological information. From the Victorian CP Register, 2956 individuals (1658 males, 1298 females), born 1970-2003, with spastic CP were identified. The proportions with each topographical pattern were analysed overall and by gestational age. Binary logistic regression analysis was used to assess temporal trends. For the review, data were systematically collected on topographical patterns from 27 registries. Estimates of heterogeneity were obtained, overall and by region, reporting period and definition of quadriplegia. Among individuals born <32 weeks, 48% had diplegia, whereas the proportion for children born >= 32 weeks was 24% (p < 0.001). Evidence was weak for a temporal trend in the relative proportions of USCP and BSCP (p = 0.038), but much clearer for an increase in the proportion of spastic diplegia relative to quadriplegia (p < 0.001). The review revealed wide variations across studies in the proportion of diplegia (range 34-90%) and BSCP (range 51-86%). These findings argue against a topographical classification based solely on laterality. PMID- 21624821 TI - The impact of background antiepileptic drugs on the efficacy and safety of pregabalin in treating partial-onset seizures: a post hoc analysis of combined clinical trials. AB - Pregabalin is used as adjunctive treatment for partial-onset seizures and is often combined with multiple antiepileptic drugs (AEDs) from different classes. The objectives of this post hoc analysis were to evaluate the efficacy and safety of pregabalin when added to different AED regimens and to identify specific AED combinations that in conjunction with pregabalin yield high responder rates. Data from six double-blind, randomized studies of pregabalin in patients with partial onset seizures were pooled for analysis (N=1775). When the treatment groups (placebo, 150mg, 300mg, 600mg, and flexible dose) were stratified by the number of concomitant AEDs (one, two or three or more), modeling results suggested that the magnitude of improvement on either >=50% responder rate or mean response ratio remained consistent regardless of the number of concomitant AEDs. Adverse events were typical of pregabalin and, in general, did not vary as the number of concomitant AEDs increased. A cluster analysis was performed to identify possible combinations of AEDs that yielded high >=50% responder rates. The majority of patients (>90%) fell within two clusters that yielded high responder rates, while <10% of the patients fell within two clusters that yielded low responder rates. Numerous AED combinations, ranging from 6 to 11, occurred within each cluster. In summary, pregabalin provided a consistent improvement in seizure reduction and comparable tolerability in patients with partial-onset epilepsy regardless of the number of concomitant AEDs. PMID- 21624820 TI - Autoantibodies with enzymatic properties in human autoimmune diseases. AB - Immunoglobulins (Ig) or antibodies are heavy plasma proteins, with sugar chains added to amino acid residues by N-linked glycosylation and occasionally by O linked glycosylation. The versatility of antibodies is demonstrated by the various functions that they mediate such as neutralization, agglutination, fixation with activation of complement and activation of effector cells. In addition to this plethora of functions, some antibodies express enzymatic activity. Antibodies endowed with enzymatic properties have been described in human autoimmune manifestations in a variety of disorders such as autoimmune thyroiditis, systemic erythematosus (SLE), scleroderma, rheumatoid arthritis (RA), multiple sclerosis (MS) and acquired hemophilia (AH). Antibodies isolated from these conditions were able to specifically hydrolyze thyroglobulin, DNA, RNA, myelin basic protein (MBP), and factor VIII (FVIII) or factor IX (FIX), respectively. The therapeutic relevance of these findings is discussed. PMID- 21624822 TI - Acute psychopathology as a predictor of global functioning in patients with ICD 10 non-affective psychosis: a prospective study in 11 European countries. AB - This prospective analysis aimed to study the influence of psychopathological dimensions on the global functioning of persons suffering from psychotic disorders, taking into account the role of a broad range of potential confounders. A large international cohort (n=1888) with ICD-10 non-affective psychosis was evaluated both at baseline during a hospital admission and three months after discharge. Trained interviewers administered a global functioning scale (GAF) and a psychopathological scale (BPRS) at baseline and follow-up). Baseline BPRS psychopathological dimensions were extracted using Principal Component Analysis. Results of multiple linear regression analyses demonstrated that affective symptoms (depressive or manic) prospectively predict a better global functioning, whilst agitation/cognitive symptoms determined poorer global functioning. Other predictors showing an independent effect on better global functioning were medication compliance, country of residence, female gender, married or coupled status, younger age and having a diagnosis of schizoaffective disorder rather than schizophrenia or other ICD-10 psychosis. A predicting model for global functioning in patients with psychosis is provided, showing that assessment of affective and agitation/cognitive symptoms should be emphasised during admission as they can be more informative than positive/negative symptoms in prospectively planning follow-up care that is geared towards a better functional recovery. PMID- 21624823 TI - Differential effects of chronic cannabis use on preattentional cognitive functioning in abstinent schizophrenic patients and healthy subjects. AB - INTRODUCTION: A number of studies indicate a higher risk for psychosis as well as for neurocognitive deficits in healthy cannabis users. However, little is known about the impact of cannabis use on outcome in schizophrenia. In fact, there is growing evidence that cannabis-using schizophrenic patients may show preserved or even better neurocognitive performance compared to schizophrenic non-users. METHODS: We measured mismatch negativity (MMN) to investigate preattentional neurocognitive functioning in long-term abstinent chronic cannabis users with (SZCA n=27) and without schizophrenia (COCA n=32) compared to schizophrenic patients (SZ n=26) and healthy controls (CO n=34) without any chronic drug use. RESULTS: Healthy cannabis users showed reduced frontal MMN compared to controls (p=0.036). In contrast, cannabis-using schizophrenic patients showed increased frontal MMN compared to schizophrenic patients without cannabis use (p=0.038). Comparing non-cannabis users, schizophrenic patients showed reduced frontal MMN (p=0.001). No significant differences were found between CO and SZCA (p=0.27), and COCA and SZCA (p=0.50). CONCLUSION: Results suggest that chronic cannabis use may have different effects on preattentional neurocognitive functioning in schizophrenic patients when compared to healthy subjects. This may be related to preexisting differences in the endocannabinoid system between schizophrenic patients and healthy subjects. However, due to the naturalistic design of the study, the results must be interpreted with caution. PMID- 21624824 TI - The evolution of antipsychotic switch and polypharmacy in natural practice--a longitudinal perspective. AB - OBJECTIVE: Most patients with schizophrenia first start with a single antipsychotic, and yet most finally end up 'switching' or using 'polypharmacy'. The objective of this study was to examine the evolution of antipsychotic switch and polypharmacy in the real-world from a longitudinal perspective. METHODS: A systematic review of longitudinal antipsychotic prescriptions in 300 patients with schizophrenia (ICD-10) for up to 2 years after their first visit to one of the 4 participating psychiatric clinics in Tokyo, Japan between January, 2007 and June, 2008, was conducted. Reasons for prescription change were also examined. The evolution of switching and polypharmacy was studied, and prescribed doses were compared to suggested dose ranges by the Texas Medication Algorithm Project (TMAP). RESULTS: 208 patients started their antipsychotic treatment with monotherapy. 34.1% of the patients gave up monotherapy with an initial antipsychotic to move to antipsychotic switch (27.4%) and/or polypharmacy (17.8%) within 2 years. The main reason for antipsychotic switch was 'ineffectiveness'; interestingly, this happened despite the fact that the monotherapy dose was below the recommended range in 47.4% of the antipsychotic switch. In a subgroup of 100 patients who started as antipsychotic-free, 2-year prevalence rates of switching and antipsychotic polypharmacy were 27.0% and 18.0%, respectively, and polypharmacy was resorted to after a median of 1 antipsychotic had been tried for 84 days (median). CONCLUSIONS: These findings raise a concern that physicians may perform an antipsychotic switch without exploring the entire dose range and resort to antipsychotic polypharmacy without trying an adequate number of antipsychotics. PMID- 21624825 TI - Time trends in occupational asthma in Belgium. AB - OBJECTIVES: There is little information on the changes in the pattern of occupational asthma (OA) over time. The objective of this study was to evaluate the time trends in the incidence and causes of immunological OA in Belgium using workers' compensation data. METHODS: Cases of OA were identified through a retrospective review of all claims submitted by salaried workers to the Belgian Workers' Compensation Board from 1993 to 2002. The likelihood of OA was categorized as definite, probable, or possible based upon the results of from diagnostic procedures. Time trends were evaluated using Spearman's rank correlation for crude numbers of cases and through the negative binomial regression for incidence rates of OA per industry. RESULTS: From a total of 1852 claims, 971 were categorized as having definite, probable, or possible OA. There was a downward trend in the annual number of cases due to the main causes of OA (i.e. flour, isocyanates, woods, and enzymes) and an apparent increase in cases of latex-induced OA. The estimated average annual incidence of OA was 29.4 (95% CI: 27.6-31.3) new cases per million salaried workers during the 1993-2002 period. There was a significant decline in the overall incidence rate of OA throughout the study period from 35.5 new cases per million salaried workers in 1993 to 25.8 in 2002. CONCLUSIONS: These compensation-based data indicate that there has been a global downward trend in the incidence of OA during the nineties, although the factors that determined this reduced incidence should be further investigated. PMID- 21624826 TI - Contextual modulation of behavioral choice. AB - We review the influence of context on behavioral choice. Context can refer to external (environmental) factors such as the season or presence of predators and it can also refer to the internal or behavioral state of an animal. Usually, animals make decisions in the midst of other ongoing behaviors. We discuss recent findings on the impact of both types of contexts, focusing on how context gets encoded at the intersection between the sensory and motor systems, emphasizing the role of neuromodulators. We also review recent technological advances that have made feasible the exploration of neural correlates of decision making in freely moving, behaving animals. PMID- 21624827 TI - Identification of crucial yeast inhibitors in bio-ethanol and improvement of fermentation at high pH and high total solids. AB - Compounds inhibitory to enzymatic hydrolysis and fermentation are generated from neutral steam exploded corn stover in the process of producing bio-ethanol. In this study, weak acids were identified as main yeast inhibitors, while phenols and aldehyde contribute to the inhibition to a lower degree. Main weak acids in hydrolysates are acetic acid and formic acid, for which critical levels for yeast inhibition are 6 and 4g/L, respectively. The inhibitory effect of these compounds can be greatly overcome by increasing pH of hydrolysates to 6.0-9.0, but there is a risk of bacterial contamination when fermenting at high pH. The relationship of pH, total solids of hydrolysates, fermentation and contamination was studied in detail. Results indicate that the contamination by bacteria when fermenting at high pH can be prevented effectively using hydrolysates with total solids of more than 20%. Meanwhile, ethanol yield is improved significantly. PMID- 21624828 TI - Competitive biosorption of Acid Blue 25 and Acid Red 337 onto unmodified and CDAB modified biomass of Aspergillus oryzae. AB - The performance of unmodified and cetyldimethylethyl ammonium bromide (CDAB) modified nonviable Aspergillus oryzae for removal of Acid Blue 25 (AB 25) and Acid Red 337 (AR 337) was investigated in single and binary systems. In single system, the biosorption capacities of CDAB-modified biosorbent reached 160.36 and 280.39mg g(-1) for AB 25 and AR 337, respectively, which were 1.52 and 1.66 times higher than that of unmodified biosorbent. In binary system, the biosorption capacities of unmodified and CDAB-modified biosorbents for both dyes decreased significantly compared to that in single system. Relative competitiveness analysis demonstrated that there existed critical initial concentration ratio which determined the predominance of dyes during biosorption process. The biosorption of AB 25 was found to be in dominant position at initial concentration ratio of [AB 25]/[AR 337] above 0.63. Kinetic analysis indicated that intraparticle diffusion was the limiting step for biosorption of two dyes onto biosorbents. PMID- 21624829 TI - A two-stage ultrafiltration and nanofiltration process for recycling dairy wastewater. AB - A two-stage ultrafiltration and nanofiltration (UF/NF) process for the treatment of model dairy wastewater was investigated to recycle nutrients and water from the wastewater. Ultracel PLGC and NF270 membranes were found to be the most suitable for this purpose. In the first stage, protein and lipid were concentrated by the Ultracel PLGC UF membrane and could be used for algae cultivation to produce biodiesel and biofuel, and the permeate from UF was concentrated by the NF270 membrane in the second stage to obtain lactose in retentate and reusable water in permeate, while the NF retentate could be recycled for anaerobic digestion to produce biogas. With this approach, most of dairy wastewater could be recycled to produce reusable water and substrates for bioenergy production. Compared with the single NF process, this two-stage UF/NF process had a higher efficiency and less membrane fouling. PMID- 21624830 TI - Furfural production from xylose using sulfonic ion-exchange resins (Amberlyst) and simultaneous stripping with nitrogen. AB - The aim of this work deals with the development of new approaches to the production of furfural from xylose. It combines relatively cheap heterogeneous catalysts (Amberlyst 70) with simultaneous furfural stripping using nitrogen under semi-batch conditions. Nitrogen, compared to steam, does not dilute the vapor phase stream when condensed. This system allowed stripping 65% of the furfural converted from xylose and almost 100% of selectivity in the condensate. Moreover, high initial xylose loadings led to the formation of two water-furfural phases, which could reduce further purification costs. Constant liquid-vapor equilibrium along stripping could be maintained for different xylose loadings. The modeling of the experimental data was carried out in order to obtain a liquid vapor mass-transfer coefficient. This value could be used for future studies under steady-state continuous conditions in similar reaction-systems. PMID- 21624831 TI - Leaching behavior of iron from simulated landfills with different operational modes. AB - The aim of the present study was to investigate the leaching behavior of iron from simulated landfills with different operation modes, with an emphasis on the variation of iron in different oxidation state, ferrous Fe(II) and ferric Fe(III) percentage and the distribution of iron content in different landfill leachate fractions. The leaching behavior and accumulated amounts of iron leached out by leachate from conventional landfill (CL) and leachate recirculated landfill (RL) exhibited decidedly different trends except for the initial 28 days. In addition, the percentage of iron leached from CL and RL accounted 1.00% and 0.14% for the total amount in landfills, respectively. No correlations between iron and selected characteristics in leachate were found were observed in the two simulated landfills. Significant positive correlations between particulate bound iron and Fe(III) were found in the leachates from RL (R(2)=0.748) and CL (R(2)=0.833). PMID- 21624832 TI - Effects of different pretreatment strategies on corn stalk acidogenic fermentation using a microbial consortium. AB - The effects of sulfuric acid, acetic acid, aqueous ammonia, sodium hydroxide, and steam explosion pretreatments of corn stalk on organic acid production by a microbial consortium, MC1, were determined. Steam explosion resulted in a substrate that was most favorable for microbial growth and organic acid productions. The total amounts of organic acids produced by MC1 on steam exploded, sodium hydroxide, sulfuric acid, acetic acid, and aqueous ammonia pretreated corn stalk were 2.99, 2.74, 1.96, 1.45, and 2.21g/l, respectively after 3days of fermentation at 50 degrees C. The most prominent organic products during fermentation of steam-exploded corn stalks were formic (0.86g/l), acetic (0.59g/l), propanoic (0.27g/l), butanoic (0.62g/l), and lactic acid (0.64g/l) after 3days of fermentation; ethanol (0.18g/l), ethanediol (0.68g/l), and glycerin (3.06g/l) were also produced. These compounds would be suitable substrates for conversion to methane by anaerobic digestion. PMID- 21624833 TI - Reflector based chlorophyll production by Spirulina platensis through energy save mode. AB - The present study suggests that the effect of silver coated polyester film fixed in culture racks serves as a reflector of light intensity on Spirulina platensis cultivation, using of KNO(3) and urea as nitrogen sources. The use of light reflector (LR) gave light intensity of 4.8-6.0klux and the reduction in number of tube light with reflector gave 2.5klux of light intensity and its effect was studied on S. platensis. Total chlorophyll productions were observed for the cultivation at light intensity of two lights with reflector. This improvement is simple, inexpensive and saves 50% electric energy by reducing the number of lights, and thus contributes to energy conservation. PMID- 21624834 TI - Bioreactor and process design for biohydrogen production. AB - Biohydrogen is regarded as an attractive future clean energy carrier due to its high energy content and environmental-friendly conversion. It has the potential for renewable biofuel to replace current hydrogen production which rely heavily on fossil fuels. While biohydrogen production is still in the early stage of development, there have been a variety of laboratory- and pilot-scale systems developed with promising potential. This work presents a review of advances in bioreactor and bioprocess design for biohydrogen production. The state-of-the art of biohydrogen production is discussed emphasizing on production pathways, factors affecting biohydrogen production, as well as bioreactor configuration and operation. Challenges and prospects of biohydrogen production are also outlined. PMID- 21624835 TI - Odontogenic myxoma: diagnostic and therapeutic challenges in paediatric and adult patients--a case series and review of the literature. AB - INTRODUCTION: Odontogenic myxomas are benign but locally invasive tumours originating from primordial mesenchymal tooth forming tissues which do not metastasise. We present a series of two paediatric and two adult cases and focus on differences in diagnostic and therapeutic approaches between children and adults based on our own experience and a critical review of the literature. PMID- 21624836 TI - Upper airway changes in syndromic craniosynostosis patients following midface or monobloc advancement: correlation between volume changes and respiratory outcome. AB - In syndromic craniosynostosis patients, respiratory insufficiency may be a pressing indication to surgically increase the patency of the upper airway by midface or monobloc advancement. In this study the volume changes of the upper airway and the respiratory outcome following midface (Le Fort I or III) or monobloc advancement in ten syndromic craniosynostosis patients are evaluated. Pre- and postoperatively, the airway volume was measured using a semi-automatic region growing method. Respiratory data were correlated to the volume measurements. In nine patients the outcome of upper airway volume measurements correlated well to the respiratory outcome. Three of these patients showed a minimal airway volume gain or even volume loss, and no respiratory improvement was found. In one monobloc patient improvement of the respiratory outcome without an evident volume gain of the upper airway was found. The majority of patients with Le Fort III advancement showed respiratory improvement, which for the greater part correlated to the results of the volume analysis. In monobloc patients the respiratory outcomes and volume measurements were less obvious. Preoperative endoscopy of the upper airway is advocated to identify the level of obstruction in patients with residual obstructive sleep apnoea. PMID- 21624837 TI - Arterial versus portal venous embolization for induction of hepatic hypertrophy before extended right hemihepatectomy in hilar cholangiocarcinomas: a prospective randomized study. AB - PURPOSE: To assess the efficacy and safety of portal vein (PV) embolization versus hepatic artery embolization (HAE) for induction of hepatic hypertrophy before extended right hemihepatectomy in patients with hilar cholangiocarcinoma. MATERIALS AND METHODS: Fifty patients (female, n = 15; male, n = 35; age range, 37-80 y) with hilar cholangiocarcinomas who were planned to undergo extended right hemihepatectomy were prospectively included in 2003-2006. In addition to biliary decompression of the left liver, patients were randomized to undergo embolization of the right hepatic artery (with transfemoral access and polyvinyl alcohol [PVA] particles plus coils) or right PV branches (with computed tomography [CT]-guided transhepatic access and PVA particles). CT was performed before and approximately 3 weeks after embolization for volumetric assessment of the liver. RESULTS: In the HAE group, median growth of the left lateral segments was 40 mL (P < .01), with a median reduction of the whole liver of 10 mL (P = .41); adverse events were observed in two of 25 patients (8%), who each developed an abscess in the right liver lobe. In the PV embolization group, median growth of the left lateral segments was 110 mL (P < .01), with a median growth of the whole liver of 10 mL (P = .92); a subcapsular seroma occurred in one of 25 patients (4%). The median growth of the left lateral segments after PV embolization was significantly greater than after HAE (P = .004). CONCLUSIONS: Compared with HAE, PV embolization was significantly superior regarding induction of hepatic hypertrophy of the left lateral segments. PMID- 21624838 TI - High incidence of asymptomatic renal cancer in late survivors after heart transplantation. PMID- 21624839 TI - Fibroblast phenotypes and their activity are changed in the wound healing process after lung transplantation. AB - BACKGROUND: Lung transplantation (LTx) is established as a life-saving treatment in end-stage lung disease. However, long-term survival is hampered by the development of chronic rejection, almost synonymous with bronchiolitis obliterans syndrome (BOS). The rejection is characterized by deposition of extracellular matrix in small airways. Fibroblasts/myofibroblasts are the main producers of extracellular matrix molecules such as proteoglycans. This study compared fibroblast phenotype and activity in the wound healing process at different points after LTx in patients who later did, or did not, develop BOS. METHODS: Distally derived fibroblasts from patients 6 and 12 months after LTx and from healthy controls were analyzed for production of the proteoglycans versican, perlecan, biglycan, and decorin, with and without transforming growth factor (TGF)-beta(1). Fibroblast migration and proliferation were also studied. RESULTS: At 6 and 12 months after LTx, versican production was higher in fibroblasts from LTx patients (p < 0.01 p < 0.01) than from controls. Fibroblasts from patients who later developed BOS were more responsive to TGF-beta(1)-induced synthesis of versican and biglycan than patients without signs of rejection (p < 0.05). Production of perlecan and decorin was negatively correlated with fibroblast proliferation in fibroblasts at 6 months after LTx. In a more detailed case study of 2 patients, one with and one without BOS, the altered proteoglycan profile was associated with impaired lung function. CONCLUSIONS: LTx changes the phenotype of fibroblasts to a non-proliferative but extracellular matrix-producing cell due to wound healing involving TGF-beta(1). If not controlled, this may lead to development of BOS. PMID- 21624840 TI - Laparoscopic liver resection: an examination of our first 300 patients. AB - BACKGROUND: Laparoscopic liver resection is a procedure in evolution. In the last decade it has evolved from a novel procedure to a standard part of the hepatic surgeon's armamentarium. Few data exist on the development of a laparoscopic resection program. STUDY DESIGN: With IRB approval, a retrospective review of 300 consecutive laparoscopic liver resections was undertaken. To determine changing results and patterns of practice, the cohort was divided into 3 consecutive groups of 100 patients. Patient demographics, indications for operation, operative factors, and in-hospital outcomes were examined. Continuous variables were analyzed with the Kruskal-Wallis test; continuous variables were compared with Fisher's exact test. Univariate and multivariate analyses of major complications (>=grade 3) were performed using logistic regression. RESULTS: Of the 300 patients, 173 (61.6%) were female, with a median age of 54 years. There were 133 (44.3%) major resections. The median number of segments resected increased (3 vs 2, p = 0.015), as did the percentage of repeat hepatectomies (13.0% vs 2.0%, p = 0.001). At the same time, median operative time decreased (2.25 vs 3.0 hours, p < 0.001).and estimated blood loss was similar (150 mL vs 150 mL, p = 0.635). Morbidity was similar (11% vs 14%, p = 0.300), as was mortality (1% vs 3%, p = 0.625). CONCLUSIONS: Laparoscopic liver resection has evolved from a novel procedure to a vital technique in liver surgery. Our group has demonstrated the ability over time to perform more difficult resections with similar morbidity and decreased operative length. PMID- 21624841 TI - Trends in use of bariatric surgery, 2003-2008. AB - BACKGROUND: During the past decade, the field of bariatric surgery has changed dramatically. This study was intended to determine trends in the use of bariatric surgery in the United States. Data used were from the Nationwide Inpatient Sample from 2003 through 2008. STUDY DESIGN: We used ICD-9 diagnosis and procedural codes to identify all hospitalizations during which a bariatric procedure was performed for the treatment of morbid obesity between 2003 and 2008. Data were reviewed for patient characteristics, annual number of bariatric procedures, and proportion of laparoscopic cases. US Census data were used to calculate the population-based annual rate of bariatric surgery per 100,000 adults. The number of surgeons performing bariatric surgery was estimated by the number of members in the American Society for Metabolic and Bariatric Surgery. RESULTS: For the period between 2003 and 2008, the number of bariatric operations peaked in 2004 at 135,985 cases and plateaued at 124,838 cases in 2008. The annual rate of bariatric operations peaked at 63.9 procedures per 100,000 adults in 2004 and decreased to 54.2 procedures in 2008. The proportion of laparoscopic bariatric operations increased from 20.1% in 2003 to 90.2% in 2008. The number of bariatric surgeons with membership in the American Society for Metabolic and Bariatric Surgery increased from 931 to 1,819 during the 6 years studied. The in-hospital mortality rate decreased from 0.21% in 2003 to 0.10% in 2008. CONCLUSIONS: In the United States, the number of bariatric operations peaked in 2004 and plateaued thereafter. Use of the laparoscopic approach to bariatric surgery has increased to >90% of bariatric operations. In-hospital mortality continually decreased throughout the 6-year period. PMID- 21624842 TI - IGF-I and IGFBP-3 in healthy 9 month old infants from the SKOT cohort: breastfeeding, diet, and later obesity. AB - BACKGROUND: High IGF-I concentrations in infancy have been associated with later obesity but the interactions between diet, IGF-I concentrations and growth in early life are complex and involve programming of the IGF-I axis. OBJECTIVE: This paper examines how IGF-I and IGFBP-3 concentrations measured at age 9 months are related to diet and growth in infancy. DESIGN: In the Danish SKOT cohort healthy term infants were included at age 9 months with follow-up at age 18 months. Total 252 infants had a full data set and were included in the analysis. Measurements include weight, length, skinfold thickness, waist circumference, 7-d food records, and blood analysis of IGF-I, and IGFBP-3. RESULTS: Infants not being breastfed at 9 months of age (46%) had higher median IGF-I concentration than breastfed infants (51.6 vs. 44.2 ng/mL, P=0.0005) and there was a negative dose response effect of daily numbers of breastfeedings on IGF-I concentration. IGF-I concentration was negatively associated with birth weight and positively related to increase in weight, length and BMI between birth and 9 months. Between 9 months and 18 months of age increase in length was positively and increase in BMI was negatively related to IGF-I concentration. CONCLUSION: Breastfeeding has a strong negative effect on IGF-I concentrations in late infancy. Although IGF-I concentrations at 9 months of age were negatively associated with change in BMI during the following 9 months we speculate that this could reflect an early adiposity rebound and thereby an increased risk of obesity later in life. PMID- 21624843 TI - Effect of tetrahydrobiopterin on Phe/Tyr ratios and variation in Phe levels in tetrahydrobiopterin responsive PKU patients. AB - BACKGROUND: Whilst a reduction in blood phenylalanine (Phe) levels is essential in patients with PKU, a decrease in Phe/Tyrosine (Tyr) ratio and fluctuations in blood Phe levels over time have been recently associated with improved neuropsychological outcome. The aim of this study was to identify if Tetrahydrobiopterin (BH(4)) offers additional benefit based on the assumption that these 2 factors are beneficial. METHOD: Since 2002, 9 patients identified through NBS as BH(4) responsive (BH(4) group) and 25 non-responsive patients (non BH(4) group) produced a total of 1384 and 4415 samples, respectively, for analysis. Statistical analysis was performed to compare mean and median Phe levels, Tyr levels and Phe/Tyr ratios in BH(4) and non BH(4) responsive patients. RESULTS: Variations in blood Phe levels were greater in the non-BH(4) group (BH(4): median 338 MUmol/L, 95% Confidence Interval (CI) 329-346, mean: 358 MUmol/L, CI 350-366; non-BH(4): median 338 MUmol/L CI 332-344, mean: 370 MUmol/L CI 364-376). Variations in blood Tyr levels were slightly greater in the non BH(4) group: (BH(4): median 59 MUmol/L CI 58-61, mean 67 CI 66-69; non-BH(4): median 62 MUmol/L CI 61-63, mean 70 CI 69-71). The variation in Phe/Tyr ratios was greater in the non-BH(4) group (mean 6.12, CI 5.9-6.3) than in the BH(4) group (Mean 5.44, CI 5.3-5.6), particularly at blood Phe levels >600 MUmol/L. CONCLUSION: BH(4) responsive patients have smaller variations in blood Phe levels and tighter Phe/Tyr ratios than non-BH(4) responsive patients, particularly at high blood Phe levels. If decreased fluctuations in Phe levels and a decreased Phe/Tyr ratio are indeed neuro-protective, then BH(4) responsiveness is advantageous over diet alone in PKU. Neuropsychological testing in patients who have been treated with BH(4) long term may be able to ascertain the clinical benefit of these biochemical findings. PMID- 21624844 TI - Monte Carlo derivation of AAPM TG-43 dosimetric parameters for GZP6 Co-60 HDR sources. AB - Cobalt 60 source is generally available on high dose rate (HDR) afterloading equipment especially for treatment of gynecological lesions. The GZP6 remote afterloader (Nuclear Power Institute of China) utilizes (60)Co sources for treatment of intracavitary and intraluminal malignancies. In this study, the AAPM TG-43 dosimetric parameters of three sources in GZP6 system have been studied using MCNP4C Monte Carlo (MC) code; and the results are compared with other available (60)Co HDR sources. The presented parameters consist of air kerma strength, dose rate constant, radial dose function and anisotropy function. They show less than 1% uncertainty. The TG-43 based dosimetry data can be used not only to validate the dedicated treatment planning software (TPS), but also to introduce new complementary software to enhance the system performance in gynecological treatments. PMID- 21624845 TI - Registration of 3D trans-esophageal echocardiography to X-ray fluoroscopy using image-based probe tracking. AB - Two-dimensional (2D) X-ray imaging is the dominant imaging modality for cardiac interventions. However, the use of X-ray fluoroscopy alone is inadequate for the guidance of procedures that require soft-tissue information, for example, the treatment of structural heart disease. The recent availability of three dimensional (3D) trans-esophageal echocardiography (TEE) provides cardiologists with real-time 3D imaging of cardiac anatomy. Increasingly X-ray imaging is now supported by using intra-procedure 3D TEE imaging. We hypothesize that the real time co-registration and visualization of 3D TEE and X-ray fluoroscopy data will provide a powerful guidance tool for cardiologists. In this paper, we propose a novel, robust and efficient method for performing this registration. The major advantage of our method is that it does not rely on any additional tracking hardware and therefore can be deployed straightforwardly into any interventional laboratory. Our method consists of an image-based TEE probe localization algorithm and a calibration procedure. While the calibration needs to be done only once, the GPU-accelerated registration takes approximately from 2 to 15s to complete depending on the number of X-ray images used in the registration and the image resolution. The accuracy of our method was assessed using a realistic heart phantom. The target registration error (TRE) for the heart phantom was less than 2mm. In addition, we assess the accuracy and the clinical feasibility of our method using five patient datasets, two of which were acquired from cardiac electrophysiology procedures and three from trans-catheter aortic valve implantation procedures. The registration results showed our technique had mean registration errors of 1.5-4.2mm and 95% capture range of 8.7-11.4mm in terms of TRE. PMID- 21624846 TI - Color treatment in endoscopic image classification using multi-scale local color vector patterns. AB - In this work we propose a novel method to describe local texture properties within color images with the aim of automated classification of endoscopic images. In contrast to comparable Local Binary Patterns operator approaches, where the respective texture operator is almost always applied to each color channel separately, we construct a color vector field from an image. Based on this field the proposed operator computes the similarity between neighboring pixels. The resulting image descriptor is a compact 1D-histogram which we use for a classification using the k-nearest neighbors classifier. To show the usability of this operator we use it to classify magnification-endoscopic images according to the pit pattern classification scheme. Apart from that, we also show that compared to previously proposed operators we are not only able to get competitive classification results in our application scenario, but that the proposed operator is also able to outperform the other methods either in terms of speed, feature compactness, or both. PMID- 21624847 TI - Fatigue in patients undergoing chemotherapy, their self-care and the role of health professionals: a qualitative study. AB - PURPOSE: Fatigue affects most chemotherapy recipients, who commonly experience it as distressing. Previous publications showed that cancer patients frequently lacked information and education regarding fatigue; and while most engaged in self-care activities, it remained unclear why they chose specific activities or how effective those were. We explored cancer chemotherapy recipients' current perspectives regarding fatigue, with particular attention to related communication with health professionals, self-care activities, and perceived effectiveness of these measures. This study was part of a larger project that explored patients' symptom experiences during chemotherapy over three months. METHOD: In this qualitative study, 19 patients with lymphomas, breast, lung or colorectal cancer participated concurrently with treatment at a Swiss tertiary care hospital's oncology outpatient clinic. Data on patients' fatigue experiences were collected via individual interviews following their third cycle of chemotherapy. Grounded Theory methodology guided data analysis. RESULTS: At the start of their chemotherapy, health professionals informed patients that common side effects included fatigue. While all participants experienced different dimensions of fatigue, then, all were willing to endure it for the sake of an expected improvement in their conditions. Individuals' fatigue experiences depended largely on their particular life and illness circumstances. Most engaged in fatigue-related self-care activities and managed the symptom on their own. Communication with or input from health professionals was virtually absent during chemotherapy. CONCLUSIONS: Adequate and systematic information regarding fatigue and related self-care strategies need to be implemented at the beginning of chemotherapy, along with continuous assessment and individual guidance of patients throughout their treatment. PMID- 21624848 TI - Novel mutations in RAG1/2 and ADA genes in Israeli patients presenting with T-B SCID or Omenn syndrome. AB - The relative frequency of the different forms of SCID may vary in different countries. The most frequent form in Israel is the autosomal-recessive T-B- SCID or Omenn syndrome while X-linked SCID is rare. We report our immunological and genetic analyses in multicentre study of patients presenting with either T-B- SCID or Omenn syndrome. Among 16 patients, we identified 7 novel mutations in 6 patients. In the RAG1 gene we detected two novel mutations: L454Q and 469 fs 4bpdel. In the RAG 2 gene: 3 novel mutations: D65Y, G157V, and E480X. One T-B- SCID patient was found to be a compound heterozygote for new mutations in the ADA gene: W264X and R235W. Prenatal diagnosis was performed in 8 families while others refused due to religious reasons. Identification of the new mutations expands our knowledge regarding the unique features of SCID phenotype in Israel and may help the families seeking for genetic counseling. PMID- 21624849 TI - A new Italian instrument for the assessment of irritability in patients with epilepsy. AB - The purpose of the work described in this article was to analyze the psychometric properties of a new Italian instrument for the assessment of irritability in adult patients with epilepsy (I-Epi). Five hundred four patients from nine secondary and tertiary Italian centers for the care of epilepsy were recruited and interviewed. Each patient was evaluated on a series of demographic and clinical variables recorded before administration of the I-Epi and the AQ (Aggression Questionnaire), used for external validity. The final results supported the reliability and validity of the I-Epi as a measure of irritability in the adult epilepsy population. The psychometric characteristics of the I-Epi seemed fairly good. We believe that adoption of this new instrument could be very useful in both clinical and research management of patients with epilepsy. PMID- 21624850 TI - An evidence-based approach to proper diagnostic use of the electroencephalogram for suspected seizures. AB - The EEG has strengths and limitations, which must be known to use it properly. An important reason to order an EEG is to help differentiate seizures from other etiologies of paroxysmal events that may or may not be associated with loss of consciousness. Like any other diagnostic test, the EEG refines the probability of a condition, and is normally used to support the clinical diagnosis of seizures, which relies mainly on a patient's history. The EEG itself neither proves nor excludes a diagnosis of seizures, but is nevertheless essential in the diagnostic workup and proper classification of epileptic seizures and syndromes. On the basis of the most relevant articles available in the literature, this article describes an evidence-based approach to proper use of the EEG in the diagnosis of seizures in an ambulatory care setting and presents concepts such as pretest and posttest probability and sensitivity and specificity with respect to EEG as a diagnostic test. PMID- 21624851 TI - Outcomes after abdominal wall reconstruction using acellular dermal matrix: a systematic review. AB - BACKGROUND: Complex abdominal wall defects can present a significant challenge to the reconstructive surgeon. In 2003, acellular dermal matrix (ADM) was introduced as an alternative to synthetic materials with suggestions that it has improved capacity to integrate with surrounding tissues with less inclination towards infection, erosion, extrusion, adhesion formation and rejection compared with synthetic materials. This systematic review was conducted to evaluate the existing literature describing the use of ADM for abdominal wall reconstruction in an attempt to identify factors that may affect outcomes. METHODS: A review of the MEDLINE database using the search terms 'dermal matrix' and 'abdomen' or 'hernia' for prospective and retrospective human studies in English was performed. Exclusion criteria were animal studies, case reports, reviews and articles that dealt only with ADM for repair of congenital abdominal wall defects, hiatal, parastomal or inguinal hernias and enterocutaneous fistulae. Two independent reviewers performed the systematic review with the same a priori criteria, with discrepancies reconciled by the senior author. RESULTS: In October 2010, 3394 articles were identified as potentially inclusive based on the search term 'dermal matrix'. When filtered for 'abdomen' or 'hernia', 83 articles were found. Ultimately, 30 articles met criteria. No other systematic reviews, meta analyses or randomised controlled trials were identified in the existing literature. CONCLUSIONS: At this current time, there is a paucity of high-level evidence comparing ADM with other methods interfering with the ability of physicians to make data-driven recommendations on clinical indications, surgical techniques and outcomes following ADM-assisted abdominal wall reconstruction. PMID- 21624852 TI - Long-term results of finger reconstruction with microvascular toe transfers after trauma. AB - Amputation of all or most of the fingers severely disturbs the gripping function of the hand. The purpose of this study was to evaluate the long-term functional results of finger-amputation patients rehabilitated with microvascular toe transfers. Fifteen such patients (10 males, median age at injury 26 years (range 5-49 years)) were examined after a median follow-up of 18 years. Eight patients had no fingers spared by the initial trauma and the rest had at least two fingers amputated. The function of the hand was accessed subjectively (questionnaires) and objectively (tests). Further, physical parameters were measured and compared to the other healthy hand. Patients scored consistently well in the test measuring function (the Sollerman hand function test and the modified Tamai score). Activities of daily living presented on average minor difficulties. Patients regained on average 42% of grip and 84% of key pinch strength compared to the other hand. The average movement of the transfer was 28 degrees . One transfer was lost due to inability to restore permanent circulation. In addition, there were one donor and one recipient site superficial infections. We conclude that microvascular toe transfer is a reliable way to improve gripping function after amputation of fingers. Patient approval is generally good and the achieved function satisfactory. Two toe transfers should be considered for patients with no fingers left. Work-related injury may be related to decreased occupational capability. PMID- 21624853 TI - Unusual presentation of hemifacial microsomia. AB - Hemifacial microsomia is a well known yet rare collection of developmental facial defects. Almost half of all known cases exhibit additional anomalies beyond the classic malformations of the first and second brachial arches. This case is unique in that it describes a child with hemifacial microsomia and an associated oropharyngeal stenosis, nasal stenosis and a v-shaped mandibular deformity. PMID- 21624854 TI - Thumb salvage with skin grafted medial femoral corticoperiosteal free flap. AB - A 23-year-old man suffered a severe crush injury with fracture of his left thumb base phalanx and destruction of his extensor pollicis longus tendon. Immediate plate stabilisation and soft tissue coverage was performed; however, a recalcitrant pseudarthrosis developed in this compliant non-smoker despite three revisions with avascular iliac crest grafts for interphalangeal joint arthrodesis, corticocancellous bone application and hardware exchange. An attempt to improve thumb vascularity and soft tissue cover with a pedicled Foucher flap as well as through extracorporeal shock wave therapy failed. Bone healing and subsequent thumb salvage were finally achieved with a free vascularised medial femoral condyle (MFC) bone flap, which was covered directly with a full-thickness skin graft. Both bony and soft tissue healing went well and after 3 months the patient returned to work. This case demonstrates that skin grafting the periosteal surface of the MFC flap is safe and results in a thin skin-bone compositae transplant which also might be very useful for indications other than hand surgery. PMID- 21624855 TI - [Update in the diagnosis and treatment of pulmonary hypertension]. AB - The term pulmonary hypertension includes a heterogeneous group of disorders which produce an obstructive remodeling of the lung vessels characterized by a steady rise in pulmonary arterial pressure. The ensuing morbidity and mortality of this disease result from the failure of the right ventricle to compensate this increase in the after-load. The conditions to achieve an optimum treatment of pulmonary hypertension patients are: a clinical presumption and early diagnosis, an adequate stratification based on the underlying causes and the different vascular territories involved, the response to therapy and the degree of severity, the selection of the different therapeutic alternatives and their timely indication. These recommendations are aimed at the general practitioner and their objective is to facilitate the early detection, diagnosis and treatment of pulmonary hypertension patients, representing a major change in the prognosis. Moreover, as the diagnostic evaluation requires complementary tests that are not usually available in all medical centers, they can also be used as the basis to request a timely referral to institutions with the adequate complexity. PMID- 21624856 TI - The need to be aware and beware of the genetic information nondiscrimination act. AB - Genetic advancements have presented numerous discrimination predicaments to individuals, the healthcare community, and legislators at state and federal levels. Oncology nurses should be knowledgeable about the Genetic Information Nondiscrimination Act of 2008 (GINA) and its applications to clinical practice. GINA is the first federal law passed to protect U.S. citizens with inherited disorders from being treated unfairly because of their genetic makeup. Understanding the legislation known as GINA, including how it modifies existing federal laws governing health insurance coverage and employment discrimination, can assist oncology nurses in providing important education and advocating for patients and their extended families. Federal agencies that govern and enforce GINA's provisions are identified. Case situations are included to demonstrate how to apply information concerning GINA to patients with cancer and their families who are considering or have already completed genetic testing. Privacy of genetic information is a timely issue but difficult to understand; therefore, provisions of GINA should be addressed and evaluated carefully. PMID- 21624857 TI - Evidence-based practice: challenging what we think we know. PMID- 21624858 TI - Metastasis of hepatocellular carcinoma into the right atrium. PMID- 21624859 TI - Assessment and management of delirium in the older adult with cancer. AB - Recognizing delirium and its impact on older patients and their caregivers is an important aspect of oncology nursing. Delirium is the most common complication of hospital admissions for older adults and is associated with a significant amount of morbidity and mortality, prolonged hospital stays, functional decline, and decreased quality of life. Oncology nurses caring for older patients with cancer have the opportunity to recognize the onset of delirium, provide ongoing patient assessment and monitoring, and implement pharmacologic and nonpharmacologic interventions. PMID- 21624860 TI - An intervention to manage patient-reported symptoms during cancer treatment. AB - Patients with cancer receiving therapy may face a variety of complicated and stressful symptoms. Oncology nurses can advocate for patients by performing their roles as educators and comanagers of cancer-related side effects. In addition, symptom-focused education provided by oncology nurses can enable patients to administer self-care more effectively. PMID- 21624861 TI - Supporting quality and patient safety in cancer clinical trials. AB - Adverse event (AE) reporting is a critical component of all cancer clinical trials, and the National Cancer Institute's Common Terminology Criteria for Adverse EventsTM (CTCAE) is the primary system used by clinicians to describe the severity of AEs. The National Cancer Institute's Patient-Reported Outcomes version of the CTCAE (PRO-CTCAE) assesses patient self-reports of symptoms using a Web-based system that can be incorporated into all cancer clinical trials. Oncology clinical trial nurses are responsible for the protection and safety of patients enrolled in cancer trials and, therefore, should develop an understanding of PRO-CTCAE. PMID- 21624862 TI - Sleep disturbance in hospitalized recipients of stem cell transplantation. AB - Disrupted sleep is considered a patient outcome sensitive to oncology nursing care and can lead to a variety of physical and psychologic dysfunctions, such as insomnia, chronic pain, respiratory distress, obesity, stress, and anxiety. Although sleep disturbances have been studied in recipients of hematopoietic stem cell transplantations (HSCTs), these studies have not examined the acute phase of transplantation. The current study aimed to identify the level of sleep disturbance in this patient population, identify factors contributing to decreased ability to sleep for hospitalized recipients of HSCT, and compare the differences in sleep disturbance between age, gender, type of transplantation, and initial stem cell transplantation versus readmission for transplantation associated complications. Among the 69 patients studied, 26% reported clinical insomnia, as measured by the Insomnia Severity Index, and 74% had some degree of insomnia. Patient characteristics were not significantly associated with insomnia scores. Patients reported bathroom use as the most frequent reason for sleep disruption (85%). These findings suggest that sleep disturbances are common in hospitalized patients undergoing HSCT, and strategies to reduce disruptions are needed to improve patient outcomes. PMID- 21624863 TI - Cultural influences on health care in Palestine. AB - International cancer care efforts are focusing increasingly on why cancer typically is diagnosed at later stages in the developing world. In Middle Eastern countries, cancer usually is diagnosed at a more advanced stage of disease. For example, in Palestine, 42% of cases were reported stage III and 18% were stage IV. Receiving a cancer diagnosis and seeking treatment is influenced by cultural values and how the community views cancer. Healthcare providers need to understand the disparities and the influence of those disparities on health outcomes. This article is constructed using the Culture Care Theory,which depicts the importance of culture on the health behavior of the individual, and will focus on how cultural values of Palestinian patients with cancer and their families affect attitudes toward and decisions about cancer care. PMID- 21624864 TI - Glioblastoma multiforme: enhancing survival and quality of life. AB - A diagnosis of a malignant brain tumor is devastating to patients and their families. The patients' inevitable loss of independence, which can occur suddenly or gradually, is tragic, and the eventual complete dependence can be overwhelming to the family and caregivers. Glioblastoma multiforme (GBM) is the most common type of primary malignant brain tumor in adults and is associated with a disproportionately high mortality rate. The highly malignant tumor grows rapidly and has a tendency to recur through treatment. The brain itself presents a multitude of barriers to treatment, such as tumor location, accessibility for surgery, and the blood-brain barrier's natural protection. Despite access to optimal multimodality treatment, patients diagnosed with GBM have a low survival rate. Patients and families need emotional and practical support throughout the continuum of this devastating disease. Astute neurologic assessment skills and immediate and appropriate interventions are required to maintain the patient's functional status. This article provides an overview of the treatment of GBM and reviews how oncology nurses can intervene to positively improve the quality of life of patients and their families. PMID- 21624865 TI - The shift to early palliative care: a typology of illness journeys and the role of nursing. AB - For the current study, clinical observations of communication between patients, families, and clinicians during chronic, serious, or terminal illness in a cancer care trajectory were examined for patterns and trends. Five communication characteristics were concluded, which informed a typology of illness journeys experienced by patients with cancer and their families. The isolated journey characterizes an illness path in which communication about terminal prognosis and end-of-life care options are not present; communication is restricted by a curative-only approach to diagnosis as well as the structure of medical care. The rescued journey signifies a transition between curative care (hospital narrative) to noncurative care (hospice narrative), challenging patients and their families with an awareness of dying. The rescued journey allows communication about prognosis and care options, establishes productive experiences through open awareness, and affords patients and families opportunities to experience end-of life care preferences. Finally, palliative care prior to hospice provides patients and families with an illness journey more readily characterized by open awareness and community, which facilitates a comforted journey. Nurses play a pivotal role in communicating about disease progression and plans of care. The typology presented can inform a structured communication curriculum for nurses and assist in the implementation of early palliative care. PMID- 21624866 TI - Anticipatory coping: taking control of hair loss. AB - Many women consider hair loss to be one of the most difficult and feared side effects of chemotherapy treatments. On learning they will likely lose their hair, women immediately begin anticipating the event and its impact on themselves and others. Anticipation of an unwanted event can lead to greater anxiety, fear, or depression, particularly if women see or find no options for gaining some sense of control. Anticipatory coping is the process of anticipation and preparation for an expected altered appearance. By researching options, making deliberate choices, and taking specific actions to determine their appearance without hair, women gain a greater sense of control of their changing appearance. This greater sense of control may ease or lessen feelings of fear and depression and help women to take control of other aspects of their lives that contribute to a greater quality of life during cancer treatment. Oncology nurses are in a unique position to help women turn their anxiety about hair loss into an anticipatory coping process, one that increases women's sense of control over some of the outward changes taking place in their bodies and empowers them to make proactive choices regarding their overall response to cancer. PMID- 21624867 TI - Are patients with breast cancer satisfied with their decision making?: a comparison over time. AB - Choosing between lumpectomy with radiation versus mastectomy is difficult for women with early-stage breast cancer, and doubt can decrease women's confidence and satisfaction. As a result, the current study surveyed satisfaction before and after surgery in a convenience sample of women with early-stage breast cancer from a single practice. All women received either total mastectomy or lumpectomy plus radiation based on their informed choice of surgical options. The surgeon and the principal investigator educated patients about both surgeries at the time of consent. Participants answered a survey about satisfaction with their decision making before their chosen surgical procedure and again by telephone six months later. Participants felt that they had made an informed choice at the time of decision (87%) and at follow-up (93%). In addition, most women were satisfied with their choice of surgical procedure at time of decision (87%) as well as six months after surgery (96%). This study allowed women to significantly participate in their care through surgical decision making, which improved satisfaction. Nurses are uniquely positioned to support women with early-stage breast cancer in their decision-making process. PMID- 21624868 TI - Accessing implanted ports: still a source of controversy. AB - In delivering safe care to their patients, oncology nurses should be familiar with the proper use and maintenance of implanted ports. Yet, despite the ubiquitous presence of implanted ports, accessing techniques and patency maintenance remain controversial. Additional research is needed to establish practices based on evidence. PMID- 21624869 TI - Esophageal cancer and palliation of dysphagia. AB - Progressive dysphagia is the most common presenting symptom of esophageal cancer but also may occur as a side effect of treatment. Evaluation of patients' dysphagia includes determining its cause and assessing the severity. Several palliative options are available for dysphagia; the clinical situation, local expertise, and cost effectiveness help determine the appropriate treatment modality. PMID- 21624870 TI - Genetic information and discrimination: a policy analysis. AB - Because of the Human Genome Project, nurses increasingly must understand how genetics impact health and treatment decisions. Although the sequencing of the human genome was crucial, the next step is advancing the understanding of genomics, interpreting genetic information, and using that information to improve health care, particularly for patients with cancer. Oncology nurses are in a position to educate and provide appropriate counseling to patients unsure about genetic testing. PMID- 21624871 TI - Carbamazepine for acute and chronic pain in adults. PMID- 21624872 TI - Nursing's healing touch. PMID- 21624873 TI - Revision of the 'missed pill' rules. PMID- 21624874 TI - Development and validation of a pictorial nausea rating scale for children. AB - OBJECTIVE: The lack of a widely used, validated measure limits pediatric nausea management. The goal of this study was to create and validate a pictorial scale with regular incremental levels between scores depicting increasing nausea intensity. METHODS: A pictorial nausea scale of 0 to 10 with 6 faces (the Baxter Retching Faces [BARF] scale) was developed in 3 stages. The BARF scale was validated in emergency department patients with vomiting and in healthy patients undergoing day surgery procedures. Patients were presented with visual analog scales for nausea and pain, the pictorial Faces Pain Scale-Revised, and the BARF scale. Patients receiving opioid analgesics or antiemetic agents had their pain and nausea assessed before and 30 minutes after therapy. Spearman's rho correlation coefficients were calculated. A Wilcoxon matched-pair rank test compared pain and nausea scores before and after antiemetic therapy. RESULTS: Thirty oncology patients and 15 nurses participated in the development of the scale, and 127 patients (52, emergency department; 75, day surgery) ages 7 to 18 years participated in the validation. The Spearman rho correlation coefficient of the first paired BARF and visual analog scale for nausea scores was 0.93. Visual analog scales for nausea and BARF scores were significantly higher in patients requiring antiemetic agents (P = .0001) and decreased significantly after treatment (P = .0002), while posttreatment VAS (P = .20) and FPSR scores (P =.47) for patients receiving only antiemetic agents did not [corrected]. CONCLUSIONS: We describe the development of a pictorial scale with beginning evidence of construct validity for a self-report assessment of the severity of pediatric nausea. The scale had convergent and discriminant validity, along with an ability to detect change after treatment. PMID- 21624875 TI - Association of interpersonal violence with self-reported history of head injury. AB - OBJECTIVE: The purpose of this study was to examine differences in interpersonal violence among individuals who reported a head injury compared with those who did not report a head injury. METHODS: We used data from an 8-year longitudinal study of youth selected by their grade point average to study those at-risk for high school dropout in 4 public high schools in a Midwestern city (N = 850). Participants were followed up from mid-adolescence to the transition into young adulthood. One-way analyses of variance were used to test for differences in levels of interpersonal violence, and repeated measures multivariate analyses of variance were used to assess differences in levels of violence over time among participants based on reports of head injury. A series of multivariate regression analyses examined whether head injury was associated with subsequent violent behavior. RESULTS: Participants who had ever experienced a head injury before young adulthood reported more interpersonal violence in young adulthood than participants who had never had a head injury. In multivariate analyses, respondents who had a head injury in the past year reported more subsequent interpersonal violence than respondents who had not had a head injury. CONCLUSION: Our findings support other studies that link history of head injury to later interpersonal violence. PMID- 21624876 TI - Decision-making around gastrostomy-feeding in children with neurologic disabilities. AB - OBJECTIVE: The objective of this study was to understand the decision-making experiences, perspectives, and beliefs of parents of children with a neurologic disability around gastrostomy tube-feeding. DESIGN AND METHODS: We conducted a systematic review of qualitative studies to explore the experiences of parents of children with a neurologic disability around gastrostomy-feeding. We searched 5 electronic databases from inception to July 2010. Two authors independently selected articles and extracted data. Concepts and themes relevant to decision making were constructed by using thematic analysis. RESULTS: Eleven studies were selected in which experiences relevant to decision-making were reported. The decision-making process was characterized by decisional conflict. Concepts important to understanding conflict were categorized under 3 themes: values; context; and process. The value and meaning of feeding by mouth and, in contrast, with a gastrostomy tube was the dominant theme that led to internal distress for parents in decision. Feeding by mouth was seen as an enjoyable activity, an important social process, but also a struggle. Gastrostomy-feeding represented a loss of normality, a sign of disability, and a disruption of maternal nurturing and bonding. Context (child and family characteristics) and process (information sharing and support) modified the decision-making experience. CONCLUSIONS: Values associated with gastrostomy-feeding and feeding by mouth, the context of child and family, and the process of decision-making facilitated by the health care system shape parental experiences and decisional conflict. This framework will help guide interventions, such as patient decision aids, that are aimed at improving parental decision-making. PMID- 21624877 TI - Newborn screening for fragile x syndrome: do we care what parents think? PMID- 21624879 TI - Policy statement--principles of pediatric patient safety: reducing harm due to medical care. AB - Pediatricians are rendering care in an environment that is increasingly complex, which results in multiple opportunities to cause unintended harm. National awareness of patient safety risks has grown in the 10 years since the Institute of Medicine published its report To Err Is Human, and patients and society as a whole continue to challenge health care providers to examine their practices and implement safety solutions. The depth and breadth of harm incurred by the practice of medicine is still being defined as reports continue to uncover a variety of avoidable errors, from those that involve specific high-risk medications to those that are more generalizable, such as patient misidentification. Pediatricians in all venues must have a working knowledge of patient-safety language, advocate for best practices that attend to risks that are unique to children, identify and support a culture of safety, and lead efforts to eliminate avoidable harm in any setting in which medical care is rendered to children. PMID- 21624878 TI - The effect of maternity leave length and time of return to work on breastfeeding. AB - OBJECTIVE: We investigated the effect of maternity leave length and time of first return to work on breastfeeding. METHODS: Data were from the Early Childhood Longitudinal Study-Birth Cohort. Restricting our sample to singletons whose biological mothers were the respondents at the 9-month interview and worked in the 12 months before delivery (N = 6150), we classified the length of total maternity leave (weeks) as 1 to 6, 7 to 12, >= 13, and did not take; paid maternity leave (weeks) as 0, 1 to 6, >= 7, and did not take; and time of return to work postpartum (weeks) as 1 to 6, 7 to 12, >= 13, and not yet returned. Analyses included chi(2) tests and multiple logistic regressions. RESULTS: In our study population, 69.4% initiated breastfeeding with positive variation by both total and paid maternity leave length, and time of return to work. In adjusted analyses, neither total nor paid maternity leave length had any impact on breastfeeding initiation or duration. Compared with those returning to work within 1 to 6 weeks, women who had not yet returned to work had a greater odds of initiating breastfeeding (odds ratio [OR]: 1.46 [1.08-1.97]; risk ratios [RR]: 1.13 [1.03-1.22]), continuing any breastfeeding beyond 6 months (OR: 1.41 [0.87 2.27]; RR: 1.25 [0.91-1.61]), and predominant breastfeeding beyond 3 months (OR: 2.01 [1.06-3.80]; RR: 1.70 [1.05-2.53]). Women who returned to work at or after 13 weeks postpartum had higher odds of predominantly breastfeeding beyond 3 months (OR: 2.54 [1.51-4.27]; RR: 1.99 [1.38-2.69]). CONCLUSION: If new mothers delay their time of return to work, then duration of breastfeeding among US mothers may lengthen. PMID- 21624880 TI - Early administration of intratracheal surfactant (calfactant) after hydrocarbon aspiration. AB - Hydrocarbon ingestions account for a substantial number of accidental poisonings; when aspirated, it can lead to severe pneumonitis. Treatment for severe pneumonitis is generally supportive, and outcomes are frequently poor. We report here the case of a 19-month-old girl who was treated successfully with early administration of exogenous surfactant for acute respiratory distress syndrome secondary to aspiration of lamp oil. Approximately 7 hours after aspiration, she required mechanical ventilation and had an oxygenation index (OI) of 13.2. Approximately 10 hours after ingestion, exogenous surfactant (calfactant) was instilled intratracheally, after which her OI improved markedly to 4.3. She received a second dose ~19 hours after ingestion, after which her OI remained at <5 and she was progressively weaned from mechanical ventilation. She was extubated 64 hours after the ingestion with no residual lung disease. This case illustrates the importance of considering exogenous surfactant therapy early in the course of acute respiratory failure secondary to hydrocarbon aspiration. Because of the putative mechanisms of lung injury involved in hydrocarbon aspiration, surfactant-replacement therapy is a reasonable therapeutic intervention based on pathophysiologic rationale. PMID- 21624881 TI - Parents' decisions to screen newborns for FMR1 gene expansions in a pilot research project. AB - OBJECTIVE: The goal of this study was to document rates of parental consent in a pilot study of newborn screening for FMR1 gene expansions, examine demographic characteristics of mothers who consented or declined, describe the reasons for their decision, and discuss ethical and social aspects of the consent process. METHODS: A brief survey was used to record basic demographic data from mothers and an open-ended question was used to elicit parents' reasons for accepting or declining screening. A descriptive analysis was conducted on the number of mothers who consented to or declined screening, and a logistic regression model predicted mothers' likelihood to agree to screening based on demographic characteristics. Reasons for decisions were analyzed using content analysis. The study was conducted at University of North Carolina Hospitals. A total of 2137 mothers were approached. RESULTS: The uptake rate for couples was 63%. Acceptance rates varied by race/ethnicity, with black respondents being less likely to accept screening. Primary reasons for accepting were "to know," "belief in research," and "the test was minimal/no risk." Reasons for declining included not wanting to know or worry, not being a good time, and issues with testing children or with genetic tests. CONCLUSIONS: Findings demonstrate that a majority of parents accepted newborn screening for FMR1 gene expansions, but decision rates and reasons for accepting or declining varied in part as a function of race/ethnicity and in part as a function of what parents most valued or feared in their assessment of risks and benefits. PMID- 21624882 TI - Sports drinks and energy drinks for children and adolescents: are they appropriate? AB - Sports and energy drinks are being marketed to children and adolescents for a wide variety of inappropriate uses. Sports drinks and energy drinks are significantly different products, and the terms should not be used interchangeably. The primary objectives of this clinical report are to define the ingredients of sports and energy drinks, categorize the similarities and differences between the products, and discuss misuses and abuses. Secondary objectives are to encourage screening during annual physical examinations for sports and energy drink use, to understand the reasons why youth consumption is widespread, and to improve education aimed at decreasing or eliminating the inappropriate use of these beverages by children and adolescents. Rigorous review and analysis of the literature reveal that caffeine and other stimulant substances contained in energy drinks have no place in the diet of children and adolescents. Furthermore, frequent or excessive intake of caloric sports drinks can substantially increase the risk for overweight or obesity in children and adolescents. Discussion regarding the appropriate use of sports drinks in the youth athlete who participates regularly in endurance or high-intensity sports and vigorous physical activity is beyond the scope of this report. PMID- 21624883 TI - Immunogenicity and safety of H influenzae type b-N meningitidis C/Y conjugate vaccine in infants. AB - BACKGROUND: Meningococcal disease incidence is highest in children younger than 2 years of age, yet there is no US-licensed vaccine for this age group. A phase III study evaluated the immunogenicity and safety of an investigational Haemophilus influenzae type b (Hib)-Neisseria meningitidis serogroups C and Y-tetanus toxoid conjugate vaccine (HibMenCY). MATERIALS AND METHODS: A total of 4180 infants were randomly assigned to receive the HibMenCY at the ages of 2, 4, 6, and 12 to 15 months or the licensed Hib tetanus toxoid conjugate vaccine (ActHIB) at 2, 4, and 6 months and Hib conjugated to N meningitidis outer membrane protein (PedvaxHIB) at 12 to 15 months. Routinely scheduled vaccines were coadministered. Serum bactericidal activity using human complement and anti-polyribosylribitol phosphate antibodies were assessed in 991 subjects. Local and systemic adverse reactions were recorded for 4 days after each dose. RESULTS: The percentage of HibMenCY recipients with serum bactericidal assay using human complement titers of 1:8 or higher after dose 3 was 98.8% for N meningitidis serogroup C (MenC) and 95.8% for N meningitidis serogroup Y (MenY). After dose 4, the percentages were 98.5% and 98.8%, respectively. The percentage of HibMenCY recipients with postdose 3 anti-polyribosylribitol phosphate antibody levels of >= 1.0 MUg/mL was noninferior to that of control (96.3% vs 91.2%). After dose 4, MenC and MenY serum bactericidal assay using human complement antibody titers increased 12-fold over pre-dose 4 levels. Incidence of pain, redness, and swelling at the HibMenCY injection sites tended to be lower than with Hib type b after the first 3 doses and after the fourth dose. Rates of systemic symptoms were similar across groups. CONCLUSIONS: The HibMenCY was immunogenic against MenC and MenY and induced anti polyribosylribitol phosphate antibody levels noninferior to those of licensed Hib conjugate vaccine. The safety profile of the HibMenCY was clinically acceptable and comparable to Hib conjugate vaccine. PMID- 21624884 TI - Early childhood caries in indigenous communities. AB - The oral health of Indigenous children of Canada (First Nations, Inuit, and Metis) and the United States (American Indian, Alaska Native) is a major child health issue: there is a high prevalence of early childhood caries (ECC) and resulting adverse health effects in this community, as well as high rates and costs of restorative and surgical treatments under general anesthesia. ECC is an infectious disease that is influenced by multiple factors, including socioeconomic determinants, and requires a combination of approaches for improvement. This statement includes recommendations for preventive oral health and clinical care for young infants and pregnant women by primary health care providers, community-based health-promotion initiatives, oral health workforce and access issues, and advocacy for community water fluoridation and fluoride varnish program access. Further community-based research on the epidemiology, prevention, management, and microbiology of ECC in Indigenous communities would be beneficial. PMID- 21624885 TI - Early childhood development of late-preterm infants: a systematic review. AB - CONTEXT: Late-preterm infants (LPIs) born at 34 to 36 weeks' gestation are increasingly regarded as being at risk for adverse developmental outcomes. To date, the early childhood development of LPIs has not been systematically considered. OBJECTIVE: To undertake a broad examination of literature relating to early childhood development at the ages of 1 to 7 years of LPIs born at 34 to 36 weeks' gestation. METHODS: We conducted a systematic review of early childhood outcomes in LPIs by using 9 electronic databases (January 1980 to March 2010). Bibliographies were reviewed. After examination of abstracts, ineligible studies were excluded. A specifically designed data-extraction form was used. The methodologic quality of included studies was assessed by using well-documented quality-appraisal guidelines. RESULTS: Of 4581 studies, 10 (3 prospective and 7 retrospective cohort) were included. Studies were heterogeneous, and poorer outcomes were reported among LPIs in relation to neurodevelopmental disabilities, educational ability, early-intervention requirements, medical disabilities, and physical growth in comparison to term-born children. No identified study used healthy nonadmitted LPIs as a comparison group for admitted LPIs. CONCLUSIONS: Evidence suggests that LPIs are at increased risk of adverse developmental outcomes and academic difficulties up to 7 years of age in comparison to term infants. An infant control group matched for gestational age has not been used; thus, for LPIs, the effect of neonatal admission on longer-term outcomes has not been fully explored. Systematic measurement of early childhood outcomes is lacking, and focused long-term follow-up studies are needed to investigate early childhood development after late-preterm birth. PMID- 21624886 TI - Using video simulated presence to reduce resistance to care and increase participation of adults with dementia. AB - BACKGROUND: Cognitive and functional decline in dementia generally impairs performance of basic care activities. Staff assistance during these activities frequently results in confusion, anxiety, and distress, expressed through resistance to care (RTC). METHODS: A single-system ABA withdrawal design (n = 1) evaluated the effect of video-simulated presence (VSP) for decreasing RTC and increasing participation. A family member pre-recorded videos for use during episodes of RTC, in which the family member spoke directly to the participant to encourage participation. RESULTS: Introduction of the VSP significantly reduced RTC during the basic care tasks of feeding and talking medication. This effect was reversed when the intervention was withdrawn. Participation increased following VSP, demonstrating clear trends toward clinical significance. CONCLUSIONS: This person-centered intervention, based on VSP of a family member, provides encouraging results for reducing RTC and increasing participation of adults with dementia in basic care tasks. PMID- 21624887 TI - BlastR--fast and accurate database searches for non-coding RNAs. AB - We present and validate BlastR, a method for efficiently and accurately searching non-coding RNAs. Our approach relies on the comparison of di-nucleotides using BlosumR, a new log-odd substitution matrix. In order to use BlosumR for comparison, we recoded RNA sequences into protein-like sequences. We then showed that BlosumR can be used along with the BlastP algorithm in order to search non coding RNA sequences. Using Rfam as a gold standard, we benchmarked this approach and show BlastR to be more sensitive than BlastN. We also show that BlastR is both faster and more sensitive than BlastP used with a single nucleotide log-odd substitution matrix. BlastR, when used in combination with WU-BlastP, is about 5% more accurate than WU-BlastN and about 50 times slower. The approach shown here is equally effective when combined with the NCBI-Blast package. The software is an open source freeware available from www.tcoffee.org/blastr.html. PMID- 21624888 TI - SwissDock, a protein-small molecule docking web service based on EADock DSS. AB - Most life science processes involve, at the atomic scale, recognition between two molecules. The prediction of such interactions at the molecular level, by so called docking software, is a non-trivial task. Docking programs have a wide range of applications ranging from protein engineering to drug design. This article presents SwissDock, a web server dedicated to the docking of small molecules on target proteins. It is based on the EADock DSS engine, combined with setup scripts for curating common problems and for preparing both the target protein and the ligand input files. An efficient Ajax/HTML interface was designed and implemented so that scientists can easily submit dockings and retrieve the predicted complexes. For automated docking tasks, a programmatic SOAP interface has been set up and template programs can be downloaded in Perl, Python and PHP. The web site also provides an access to a database of manually curated complexes, based on the Ligand Protein Database. A wiki and a forum are available to the community to promote interactions between users. The SwissDock web site is available online at http://www.swissdock.ch. We believe it constitutes a step toward generalizing the use of docking tools beyond the traditional molecular modeling community. PMID- 21624889 TI - R3D-BLAST: a search tool for similar RNA 3D substructures. AB - R3D-BLAST is a BLAST-like search tool that allows the user to quickly and accurately search against the PDB for RNA structures sharing similar substructures with a specified query RNA structure. The basic idea behind R3D BLAST is that all the RNA 3D structures deposited in the PDB are first encoded as 1D structural sequences using a structural alphabet of 23 distinct nucleotide conformations, and BLAST is then applied to these 1D structural sequences to search for those RNA substructures whose 1D structural sequences are similar to that of the query RNA substructure. R3D-BLAST takes as input an RNA 3D structure in the PDB format and outputs all substructures of the hits similar to that of the query with a graphical display to show their structural superposition. In addition, each RNA substructure hit found by R3D-BLAST has an associated E-value to measure its statistical significance. R3D-BLAST is now available online at http://genome.cs.nthu.edu.tw/R3D-BLAST/ for public access. PMID- 21624890 TI - ADGO 2.0: interpreting microarray data and list of genes using composite annotations. AB - ADGO 2.0 is a web-based tool that provides composite interpretations for microarray data comparing two sample groups as well as lists of genes from diverse sources of biological information. Some other tools also incorporate composite annotations solely for interpreting lists of genes but usually provide highly redundant information. This new version has the following additional features: first, it provides multiple gene set analysis methods for microarray inputs as well as enrichment analyses for lists of genes. Second, it screens redundant composite annotations when generating and prioritizing them. Third, it incorporates union and subtracted sets as well as intersection sets. Lastly, users can upload their own gene sets (e.g. predicted miRNA targets) to generate and analyze new composite sets. The first two features are unique to ADGO 2.0. Using our tool, we demonstrate analyses of a microarray dataset and a list of genes for T-cell differentiation. The new ADGO is available at http://www.btool.org/ADGO2. PMID- 21624891 TI - GLAMM: Genome-Linked Application for Metabolic Maps. AB - The Genome-Linked Application for Metabolic Maps (GLAMM) is a unified web interface for visualizing metabolic networks, reconstructing metabolic networks from annotated genome data, visualizing experimental data in the context of metabolic networks and investigating the construction of novel, transgenic pathways. This simple, user-friendly interface is tightly integrated with the comparative genomics tools of MicrobesOnline [Dehal et al. (2010) Nucleic Acids Research, 38, D396-D400]. GLAMM is available for free to the scientific community at glamm.lbl.gov. PMID- 21624892 TI - EBP1 and DRBP76/NF90 binding proteins are included in the major histocompatibility complex class II RNA operon. AB - Major histocompatibility complex class II mRNAs encode heterodimeric proteins involved in the presentation of exogenous antigens during an immune response. Their 3'UTRs bind a protein complex in which we identified two factors: EBP1, an ErbB3 receptor-binding protein and DRBP76, a double-stranded RNA binding nuclear protein, also known as nuclear factor 90 (NF90). Both are well-characterized regulatory factors of several mRNA molecules processing. Using either EBP1 or DRBP76/NF90-specific knockdown experiments, we established that the two proteins play a role in regulating the expression of HLA-DRA, HLA-DRB1 and HLA-DQA1 mRNAs levels. Our study represents the first indication of the existence of a functional unit that includes different transcripts involved in the adaptive immune response. We propose that the concept of 'RNA operon' may be suitable for our system in which MHCII mRNAs are modulated via interaction of their 3'UTR with same proteins. PMID- 21624893 TI - Prevalence and risk factors for coronary artery calcification following kidney transplantation for systemic lupus erythematosus. AB - OBJECTIVES: Patients with SLE who undergo kidney transplantation are at increased risk of premature cardiovascular disease. The current study aimed to investigate the prevalence of coronary artery calcification in transplanted SLE patients without coronary symptoms and to explore risk factors associated with coronary atherosclerosis. METHODS: This was a cross-sectional study in transplanted SLE patients with a functioning graft. Evaluation included laboratory tests, SLE disease activity indices, multi-slice CT for quantification of coronary calcification (Agatston score >400 = high calcification, <400 = mild to moderate calcification). Arterial and aortic stiffness was assessed by carotid-femoral pulse wave velocity (PWV). RESULTS: Thirty-nine patients were analysed, three of whom had a prior history of coronary events. Coronary artery calcification was present in 82% of patients (n = 32), with 36% (n = 14) with a high level of arterial calcification. Multivariate regression analysis showed the following factors to be independently associated with high coronary calcification: PWV (0.41, 95% CI 0.17, 0.66, P = 0.001), time since diagnosis of LN (0.66, 95% CI 0.43, 0.71, P < 0.001) and BMI (0.39, 95% CI 0.15, 0.63, P = 0.002). Immunosuppression regimen was not significantly different between groups. CONCLUSIONS: Coronary artery calcification is widespread in transplanted SLE patients despite a normal profile of conventional cardiovascular risk factors. The risk of calcification increases with disease duration, BMI and PWV in this population. PMID- 21624894 TI - Transient acute lateral wall ST elevation myocardial infarction caused by 'benign' circunflex coronary artery anomaly originating from the right coronary artery. PMID- 21624895 TI - Characterization and financial impact of implantable cardioverter-defibrillator patients without interventions 5 years after implantation. AB - BACKGROUND: Implantable cardioverter defibrillators (ICD's) are increasingly used for primary and secondary prevention of sudden cardiac death. However, data on how many ICD patients indeed receive appropriate ICD therapy during long-term follow-up is scarce. AIM: The aim of our study was to determine the number of patients without appropriate ICD therapy 5 years after ICD implantation, to identify predicting factors, to assess the occurrence of late first ICD therapy and to quantify the financial impact of ICD therapy in a real-world setting. DESIGN: Prospective observational study. METHODS: We prospectively enrolled 322 consecutive ICD patients. Baseline data were collected at implantation and patients were followed for a median of 7.3 years (IQR 5.8-9.2 years). Time to first appropriate ICD therapy (either antitachycardia pacing or cardioversion) was documented. RESULTS: Five years after implantation, 139 patients (43%) had not received appropriate ICD therapy. In multivariable analysis, a primary prevention indication and negative electrophysiological studies prior to ICD implantation were independent predictors of freedom from ICD therapy. Of the patients without ICD therapy, 5 years after implantation, 25% had experienced inappropriate ICD shocks. Two hundred and seven devices (1.5 devices per patient) were needed for the 139 patients without ICD intervention within 5 years, accounting for ? 31,784 per patient. During an additional follow-up of 3 years, 12% of the patients with unused ICD received a late first appropriate ICD therapy. CONCLUSION: About half of the ICD patients receive appropriate ICD therapy within 5 years after implantation. Furthermore, there is a significant proportion of patients receiving late first shocks after five initially uneventful years. PMID- 21624896 TI - MaizeGDB: curation and outreach go hand-in-hand. AB - First released in 1991 with the name MaizeDB, the Maize Genetics and Genomics Database, now MaizeGDB, celebrates its 20th anniversary this year. MaizeGDB has transitioned from a focus on comprehensive curation of the literature, genetic maps and stocks to a paradigm that accommodates the recent release of a reference maize genome sequence, multiple diverse maize genomes and sequence-based gene expression data sets. The MaizeGDB Team is relatively small, and relies heavily on the research community to provide data, nomenclature standards and most importantly, to recommend future directions, priorities and strategies. Key aspects of MaizeGDB's intimate interaction with the community are the co-location of curators with maize research groups in multiple locations across the USA as well as coordination with MaizeGDB's close partner, the Maize Genetics Cooperation--Stock Center. In this report, we describe how the MaizeGDB Team currently interacts with the maize research community and our plan for future interactions that will support updates to the functional and structural annotation of the B73 reference genome. PMID- 21624897 TI - Arabidopsis thaliana mutants lacking ADP-glucose pyrophosphorylase accumulate starch and wild-type ADP-glucose content: further evidence for the occurrence of important sources, other than ADP-glucose pyrophosphorylase, of ADP-glucose linked to leaf starch biosynthesis. AB - It is widely considered that ADP-glucose pyrophosphorylase (AGP) is the sole source of ADP-glucose linked to bacterial glycogen and plant starch biosynthesis. Genetic evidence that bacterial glycogen biosynthesis occurs solely by the AGP pathway has been obtained with glgC- AGP mutants. However, recent studies have shown that (i) these mutants can accumulate high levels of ADP-glucose and glycogen, and (ii) there are sources other than GlgC, of ADP-glucose linked to glycogen biosynthesis. In Arabidopsis, evidence showing that starch biosynthesis occurs solely by the AGP pathway has been obtained with the starchless adg1-1 and aps1 AGP mutants. However, mounting evidence has been compiled previewing the occurrence of more than one important ADP-glucose source in plants. In attempting to solve this 20-year-old controversy, in this work we carried out a judicious characterization of both adg1-1 and aps1. Both mutants accumulated wild-type (WT) ADP-glucose and approximately 2% of WT starch, as further confirmed by confocal fluorescence microscopic observation of iodine-stained leaves and of leaves expressing granule-bound starch synthase fused with GFP. Introduction of the sex1 mutation affecting starch breakdown into adg1-1 and aps1 increased the starch content to 8-10% of the WT starch. Furthermore, aps1 leaves exposed to microbial volatiles for 10 h accumulated approximately 60% of the WT starch. aps1 plants expressing the bacterial ADP-glucose hydrolase EcASPP in the plastid accumulated normal ADP-glucose and reduced starch when compared with aps1 plants, whereas aps1 plants expressing EcASPP in the cytosol showed reduced ADP-glucose and starch. Moreover, aps1 plants expressing bacterial AGP in the plastid accumulated WT starch and ADP-glucose. The overall data show that (i) there occur important source(s), other than AGP, of ADP-glucose linked to starch biosynthesis, and (ii) AGP is a major determinant of starch accumulation but not of intracellular ADP glucose content in Arabidopsis. PMID- 21624898 TI - A growing molecular toolbox for the functional analysis of microRNAs in Caenorhabditis elegans. AB - With the growing number of microRNAs (miRNAs) being identified each year, more innovative molecular tools are required to efficiently characterize these small RNAs in living animal systems. Caenorhabditis elegans is a powerful model to study how miRNAs regulate gene expression and control diverse biological processes during development and in the adult. Genetic strategies such as large scale miRNA deletion studies in nematodes have been used with limited success since the majority of miRNA genes do not exhibit phenotypes when individually mutated. Recent work has indicated that miRNAs function in complex regulatory networks with other small RNAs and protein-coding genes, and therefore the challenge will be to uncover these functional redundancies. The use of miRNA inhibitors such as synthetic antisense 2'-O-methyl oligoribonucleotides is emerging as a promising in vivo approach to dissect out the intricacies of miRNA regulation. PMID- 21624899 TI - SVA: software for annotating and visualizing sequenced human genomes. AB - SUMMARY: Here we present Sequence Variant Analyzer (SVA), a software tool that assigns a predicted biological function to variants identified in next-generation sequencing studies and provides a browser to visualize the variants in their genomic contexts. SVA also provides for flexible interaction with software implementing variant association tests allowing users to consider both the bioinformatic annotation of identified variants and the strength of their associations with studied traits. We illustrate the annotation features of SVA using two simple examples of sequenced genomes that harbor Mendelian mutations. AVAILABILITY AND IMPLEMENTATION: Freely available on the web at http://www.svaproject.org. PMID- 21624900 TI - Implications of variability in definition and reporting of major bleeding in randomized trials of oral P2Y12 inhibitors for acute coronary syndromes. AB - AIMS: Various definitions of major bleeding have been used to evaluate safety in randomized controlled trials of antiplatelet therapy. We compared the definitions and rates of major bleeding in phase III randomized controlled trials of oral P2Y(12) inhibitors in the management of patients with acute coronary syndromes (ACS). METHODS AND RESULTS: Electronic searches identified six phase III randomized controlled oral P2Y(12) inhibitor trials published between 2001 and 2010 involving 119 020 patients with ACS. The trials compared clopidogrel standard-dose (300-mg loading dose, 75-mg daily thereafter) vs. placebo (CURE, CLARITY-TIMI 28, COMMIT), clopidogrel standard-dose vs. prasugrel (TRITON-TIMI 38) or ticagrelor (PLATO) and clopidogrel standard-dose vs. clopidogrel double dose (600-mg loading dose, 150-mg daily for 6-days, 75-mg daily thereafter) (CURRENT-OASIS 7). Using the trial definition, major bleeding rates in patients treated with standard-dose clopidogrel ranged from 0.6% in COMMIT to 11.2% in PLATO. The contrast in bleeding rates of standard-dose clopidogrel among the trials was attenuated when using the thrombolysis in myocardial infarction (TIMI) definition for major bleeding (range 1.1-7.7%) and bleeding rates in all the trials were less than 2% when comparing 30 day rates of non-coronary artery bypass graft surgery-related TIMI major bleeding (range 0.3-1.9%). CONCLUSION: Differences in major bleeding rates between trials of P2Y(12) inhibitors in patients with ACS are minimized after standardization of bleeding definitions, timing of reporting of bleeding outcomes, and procedure rates. Interpretation of the risk of bleeding associated with different P2Y(12) inhibitors would be facilitated by a consistent approach to the definition and reporting of bleeding. PMID- 21624901 TI - An unusual heart. PMID- 21624902 TI - Non-invasive estimation of pulmonary vascular resistance with cardiac magnetic resonance. AB - AIM: To develop a cardiac magnetic resonance (CMR) method for non-invasive estimation of pulmonary vascular resistance (PVR). METHODS AND RESULTS: The study comprised 100 consecutive patients with known or suspected pulmonary hypertension (PH; 53 +/- 16 years, 73% women) who underwent same-day right heart catheterization (RHC) and CMR. Increased PVR was defined from RHC as >3 WU (n = 66, 66%). From CMR cine and phase-contrast images, right ventricular (RV) volumes and ejection fraction (RVEF), pulmonary artery (PA) flow velocities and areas, and cardiac output were quantified. The best statistical model to estimate PVR was obtained from a derivation cohort (n = 80) based on physiological plausibility and statistical criteria. Validity of the model was assessed in the remaining 20 patients (validation cohort). The CMR-derived model was: estimated PVR (in WU) = 19.38 - [4.62 * Ln PA average velocity (in cm/s)] - [0.08 * RVEF (in %)]. In the validation cohort, the correlation between invasively quantified and CMR-estimated PVR was 0.84 (P < 0.001). The mean bias between the RHC-derived and CMR-estimated PVR was -0.54 (agreement interval -6.02 to 4.94 WU). The CMR model correctly classified 18 (90%) of patients as having normal or increased PVR (area under the receiver operator characteristics curve 0.97; 95% confidence interval: 0.89-1.00). CONCLUSIONS Non-invasive estimation of PVR using CMR is feasible and may be valuable for PH diagnosis and/or follow-up. PMID- 21624903 TI - Long-term treatment with nifedipine suppresses coronary hyperconstricting responses and inflammatory changes induced by paclitaxel-eluting stent in pigs in vivo: possible involvement of Rho-kinase pathway. AB - AIMS: Accumulating evidence indicates that coronary vasoconstricting responses are enhanced at the edges of coronary segment implanted with a drug-eluting stent (DES) compared with a bare-metal stent (BMS) in humans. We have recently demonstrated that Rho-kinase pathway plays an important role in DES-induced coronary hyperconstricting responses associated with inflammatory changes in pigs in vivo. This study examined whether long-term treatment with calcium channel blocker suppresses DES-induced coronary hyperconstricting responses in pigs in vivo. METHODS AND RESULTS: Paclitaxel-eluting stent (PES) and a BMS were randomly implanted in the left coronary arteries in male domestic pigs with and without long-acting nifedipine (NIF, 4 mg/kg/day) for 4 weeks (n = 7 each). Coronary vasomotion was evaluated by quantitative coronary angiography at least 24 h after withdrawal of NIF to avoid its direct effects on coronary vasomotion. In the control group (without NIF), coronary vasoconstricting responses to serotonin (10 and 100 ug/kg, i.c.) were significantly enhanced at the PES site compared with the BMS site (P = 0.009), which were abolished by hydroxyfasudil (90 and 300 ug/kg, i.c.), a selective Rho-kinase inhibitor. The PES-induced vasoconstricting responses were significantly inhibited in the NIF group (P = 0.019). Histological examination showed that inflammatory cell accumulation and microthrombus formation were enhanced at the PES site compared with the BMS site (P < 0.05), both of which were significantly suppressed by NIF associated with reduced Rho kinase expression and activity (P < 0.05). CONCLUSION: These results indicate that long-term treatment with NIF suppresses PES-induced coronary abnormalities partly through Rho-kinase pathway inhibition in vivo. PMID- 21624904 TI - Erythema migrans in Lyme disease. PMID- 21624905 TI - United States officials propose further retreat from first-come, first-served organ donation. PMID- 21624906 TI - Defining acute renal failure. PMID- 21624907 TI - Attitudes of cancer patients, family caregivers, oncologists and members of the general public toward critical interventions at the end of life of terminally ill patients. AB - BACKGROUND: Whereas most studies have focused on euthanasia and physician assisted suicide, few have dealt comprehensively with other critical interventions administered at the end of life. We surveyed cancer patients, family caregivers, oncologists and members of the general public to determine their attitudes toward such interventions. METHODS: We administered a questionnaire to four groups about their attitudes toward five end-of-life interventions--withdrawal of futile life-sustaining treatment, active pain control, withholding of life-sustaining measures, active euthanasia and physician assisted suicide. We performed multivariable analyses to compare attitudes and to identify sociodemographic characteristics associated with the attitudes. RESULTS: A total of 3840 individuals--1242 cancer patients, 1289 family caregivers and 303 oncologists from 17 hospitals, as well as 1006 members of the general Korean population--participated in the survey. A large majority in each of the groups supported withdrawal of futile life-sustaining treatment (87.1%-94.0%) and use of active pain control (89.0%-98.4%). A smaller majority (60.8%-76.0%) supported withholding of life-sustaining treatment. About 50% of those in the patient and general population groups supported active euthanasia or physician-assisted suicide, as compared with less than 40% of the family caregivers and less than 10% of the oncologists. Higher income was significantly associated with approval of the withdrawal of futile life-sustaining treatment and the practice of active pain control. Older age, male sex and having no religion were significantly associated with approval of withholding of life-sustaining measures. Older age, male sex, having no religion and lower education level were significantly associated with approval of active euthanasia and physician-assisted suicide. INTERPRETATION: Although the various participant groups shared the same attitude toward futile and ameliorative end-of-life care (the withdrawal of futile life sustaining treatment and the use of active pain control), oncologists had a more negative attitude than those in the other groups toward the active ending of life (euthanasia and physician-assisted suicide). PMID- 21624908 TI - New Delhi metallo-beta-lactamase-1: local acquisition in Ontario, Canada, and challenges in detection. AB - New Delhi metallo-beta-lactamase-1 (NDM-1) is a recently identified metallo-beta lactamase that confers resistance to carbapenems and all other beta-lactam antibiotics, with the exception of aztreonam. NDM-1 is also associated with resistance to many other classes of antibiotics. The enzyme was first identified in organisms isolated from a patient in Sweden who had previously received medical treatment in India, but it is now recognized as endemic throughout India and Pakistan and has spread worldwide. The gene encoding NDM-1 has been found predominantly in Escherichia coli and Klebsiella pneumoniae. We describe the isolation NDM-1-producing organisms from two patients in Toronto, Ontario. To the best of our knowledge, this is the first report of an organism producing NDM-1 that was locally acquired in Canada. We also discuss the evidence that NDM-1 can affect bacterial species other than E. coli and K. pneumoniae, the limited options for treatment and the difficulty laboratories face in detecting organisms that produce NDM-1. PMID- 21624910 TI - European health systems under siege. PMID- 21624911 TI - Paramedic program reducing emergency room congestion. PMID- 21624912 TI - Drug shortage registry under discussion. PMID- 21624913 TI - Maintenance of certification. PMID- 21624914 TI - Infectious diseases of animals and plants: an interdisciplinary approach. AB - Animal and plant diseases pose a serious and continuing threat to food security, food safety, national economies, biodiversity and the rural environment. New challenges, including climate change, regulatory developments, changes in the geographical concentration and size of livestock holdings, and increasing trade make this an appropriate time to assess the state of knowledge about the impact that diseases have and the ways in which they are managed and controlled. In this paper, the case is explored for an interdisciplinary approach to studying the management of infectious animal and plant diseases. Reframing the key issues through incorporating both social and natural science research can provide a holistic understanding of disease and increase the policy relevance and impact of research. Finally, in setting out the papers in this Theme Issue, a picture of current and future animal and plant disease threats is presented. PMID- 21624915 TI - A historical synopsis of farm animal disease and public policy in twentieth century Britain. AB - The diseases suffered by British livestock, and the ways in which they were perceived and managed by farmers, vets and the state, changed considerably over the course of the twentieth century. This paper documents and analyses these changes in relation to the development of public policy. It reveals that scientific knowledge and disease demographics cannot by themselves explain the shifting boundaries of state responsibility for animal health, the diseases targeted and the preferred modes of intervention. Policies were shaped also by concerns over food security and the public's health, the state of the national and livestock economy, the interests and expertise of the veterinary profession, and prevailing agricultural policy. This paper demonstrates how, by precipitating changes to farming and trading practices, public policy could sometimes actually undermine farm animal health. Animal disease can therefore be viewed both as a stimulus to, and a consequence of, twentieth century public policy. PMID- 21624916 TI - The changing role of veterinary expertise in the food chain. AB - This paper analyses how the changing governance of animal health has impacted upon veterinary expertise and its role in providing public health benefits. It argues that the social sciences can play an important role in understanding the nature of these changes, but also that their ideas and methods are, in part, responsible for them. The paper begins by examining how veterinary expertise came to be crucial to the regulation of the food chain in the twentieth century. The relationship between the veterinary profession and the state proved mutually beneficial, allowing the state to address the problems of animal health, and the veterinary profession to become identified as central to public health and food supply. However, this relationship has been gradually eroded by the application of neoliberal management techniques to the governance of animal health. This paper traces the impact of these techniques that have caused widespread unease within and beyond the veterinary profession about the consequences for its role in maintaining the public good of animal health. In conclusion, this paper suggests that the development of the social sciences in relation to animal health could contribute more helpfully to further changes in veterinary expertise. PMID- 21624917 TI - Learning from history, predicting the future: the UK Dutch elm disease outbreak in relation to contemporary tree disease threats. AB - Expanding international trade and increased transportation are heavily implicated in the growing threat posed by invasive pathogens to biodiversity and landscapes. With trees and woodland in the UK now facing threats from a number of disease systems, this paper looks to historical experience with the Dutch elm disease (DED) epidemic of the 1970s to see what can be learned about an outbreak and attempts to prevent, manage and control it. The paper draws on an interdisciplinary investigation into the history, biology and policy of the epidemic. It presents a reconstruction based on a spatial modelling exercise underpinned by archival research and interviews with individuals involved in the attempted management of the epidemic at the time. The paper explores what, if anything, might have been done to contain the outbreak and discusses the wider lessons for plant protection. Reading across to present-day biosecurity concerns, the paper looks at the current outbreak of ramorum blight in the UK and presents an analysis of the unfolding epidemiology and policy of this more recent, and potentially very serious, disease outbreak. The paper concludes by reflecting on the continuing contemporary relevance of the DED experience at an important juncture in the evolution of plant protection policy. PMID- 21624918 TI - Endemic cattle diseases: comparative epidemiology and governance. AB - Cattle are infected by a community of endemic pathogens with different epidemiological properties that invoke different managerial and governmental responses. We present characteristics of pathogens that influence their ability to persist in the UK, and describe a qualitative framework of factors that influence the political response to a livestock disease. We develop simple transmission models for three pathogens (bovine viral diarrhoea virus, bovine herpesvirus and Mycobacterium avium spp. paratuberculosis) using observed cattle movements, and compare the outcomes to an extensive dataset. The results demonstrate that the epidemiology of the three pathogens is determined by different aspects of within- and between-farm processes, which has economic, legal and political implications for control. We consider how these pathogens, and Mycobacterium bovis (the agent of bovine tuberculosis), may be classified by the process by which they persist and by their political profile. We further consider the dynamic interaction of these classifications with pathogen prevalence and with the action taken by the government. PMID- 21624919 TI - The development, regulation and use of biopesticides for integrated pest management. AB - Over the past 50 years, crop protection has relied heavily on synthetic chemical pesticides, but their availability is now declining as a result of new legislation and the evolution of resistance in pest populations. Therefore, alternative pest management tactics are needed. Biopesticides are pest management agents based on living micro-organisms or natural products. They have proven potential for pest management and they are being used across the world. However, they are regulated by systems designed originally for chemical pesticides that have created market entry barriers by imposing burdensome costs on the biopesticide industry. There are also significant technical barriers to making biopesticides more effective. In the European Union, a greater emphasis on Integrated Pest Management (IPM) as part of agricultural policy may lead to innovations in the way that biopesticides are regulated. There are also new opportunities for developing biopesticides in IPM by combining ecological science with post-genomics technologies. The new biopesticide products that will result from this research will bring with them new regulatory and economic challenges that must be addressed through joint working between social and natural scientists, policy makers and industry. PMID- 21624920 TI - The relationship between lay and technical views of Escherichia coli O157 risk. AB - Here, we bring together and contrast lay (accessible primarily through social science methodologies) and technical (via risk assessment and epidemiological techniques) views of the risk associated with the Escherichia coli O157 pathogen using two case study areas in the Grampian region of Scotland, and North Wales. Epidemiological risk factors of contact with farm animals, visiting farms or farm fields and having a private water supply were associated with postcode districts of higher than average disease incidence in the human population. However, this was not the case for the epidemiological risk factor of consumption of beef burgers, which was independent of disease incidence in the postcode district of residence. The proportion of the population expressing a high knowledge of E. coli O157 was greatest in high-incidence disease districts compared with low incidence areas (17% cf. 7%). This supports the hypothesis that in high-disease incidence areas, residents are regularly exposed to information about the disease through local cases, the media, local social networks, etc. or perhaps that individuals are more likely to be motivated to find out about it. However, no statistically significant difference was found between high- and low-incidence postcode districts in terms of the proportion of the population expressing a high likelihood of personal risk of infection (10% cf. 14%), giving a counterintuitive difference between the technical (epidemiological and quantitative microbiological risk assessment (QMRA)) and the lay assessment of E. coli O157 risk. This suggests that lay evaluations of E. coli O157 risk reflect intuitive and experience-based estimates of the risk rather than probabilistic estimates. A generally strong correspondence was found in terms of the rank order given to potential infection pathways, with environment and foodborne infection routes dominating when comparing public understanding with technical modelling results. Two general conclusions follow from the work. First, that integrative research incorporating both lay and technical views of risk is required in order that informed decisions can be made to handle or treat the risk by the groups concerned (e.g. the public, policy makers/risk managers, etc.). Second, when communicating risk, for example, through education programmes, it is important that this process is two-way with risk managers (e.g. including Food Standards Agency officials and communications team, public health infection control and environmental health officers) both sharing information with the public and stakeholder groups, as well as incorporating public knowledge, values and context (e.g. geographical location) into risk-management decisions. PMID- 21624921 TI - Frameworks for risk communication and disease management: the case of Lyme disease and countryside users. AB - Management of zoonotic disease is necessary if countryside users are to gain benefit rather than suffer harm from their activities, and to avoid disproportionate reaction to novel threats. We introduce a conceptual framework based on the pressure-state-response model with five broad responses to disease incidence. Influencing public behaviour is one response and requires risk communication based on an integration of knowledge about the disease with an understanding of how publics respond to precautionary advice. A second framework emphasizes how risk communication involves more than information provision and should address dimensions including points-of-intervention over time, place and audience. The frameworks are developed by reference to tick-borne Lyme borreliosis (also known as Lyme disease), for which informed precautionary behaviour is particularly relevant. Interventions to influence behaviour can be directed by knowledge of spatial and temporal variation of tick abundance, what constitutes risky behaviour, how people respond to information of varying content, and an understanding of the social practices related to countryside use. The frameworks clarify the response options and help identify who is responsible for risk communication. These aspects are not consistently understood, and may result in an underestimation of the role of land-based organizations in facilitating appropriate precautionary behaviour. PMID- 21624922 TI - Uncertainties in the governance of animal disease: an interdisciplinary framework for analysis. AB - Uncertainty is an inherent feature of strategies to contain animal disease. In this paper, an interdisciplinary framework for representing strategies of containment, and analysing how uncertainties are embedded and propagated through them, is developed and illustrated. Analysis centres on persistent, periodic and emerging disease threats, with a particular focus on cryptosporidiosis, foot and mouth disease and avian influenza. Uncertainty is shown to be produced at strategic, tactical and operational levels of containment, and across the different arenas of disease prevention, anticipation and alleviation. The paper argues for more critically reflexive assessments of uncertainty in containment policy and practice. An interdisciplinary approach has an important contribution to make, but is absent from current real-world containment policy. PMID- 21624923 TI - Integrating natural and social science perspectives on plant disease risk, management and policy formulation. AB - Plant diseases threaten both food security and the botanical diversity of natural ecosystems. Substantial research effort is focused on pathogen detection and control, with detailed risk management available for many plant diseases. Risk can be assessed using analytical techniques that account for disease pressure both spatially and temporally. We suggest that such technical assessments of disease risk may not provide an adequate guide to the strategies undertaken by growers and government to manage plant disease. Instead, risk-management strategies need to account more fully for intuitive and normative responses that act to balance conflicting interests between stakeholder organizations concerned with plant diseases within the managed and natural environments. Modes of effective engagement between policy makers and stakeholders are explored in the paper, together with an assessment of such engagement in two case studies of contemporary non-indigenous diseases in one food and in one non-food sector. Finally, a model is proposed for greater integration of stakeholders in policy decisions. PMID- 21624924 TI - How to make predictions about future infectious disease risks. AB - Formal, quantitative approaches are now widely used to make predictions about the likelihood of an infectious disease outbreak, how the disease will spread, and how to control it. Several well-established methodologies are available, including risk factor analysis, risk modelling and dynamic modelling. Even so, predictive modelling is very much the 'art of the possible', which tends to drive research effort towards some areas and away from others which may be at least as important. Building on the undoubted success of quantitative modelling of the epidemiology and control of human and animal diseases such as AIDS, influenza, foot-and-mouth disease and BSE, attention needs to be paid to developing a more holistic framework that captures the role of the underlying drivers of disease risks, from demography and behaviour to land use and climate change. At the same time, there is still considerable room for improvement in how quantitative analyses and their outputs are communicated to policy makers and other stakeholders. A starting point would be generally accepted guidelines for 'good practice' for the development and the use of predictive models. PMID- 21624925 TI - Successful treatment of hepatitis B virus associated nephrotic syndrome with oral Lamivudine in a Nigerian child: a case report. AB - Hepatitis B virus is a well described cause of nephrotic syndrome (NS) worldwide, the typical lesion being membranous glomerulonephropathy. HBV associated NS has been successfully treated with intravenous alpha interferon (IFN), an anti-viral agent. In recent times there have been reports of treatment with lamivudine, an orally administered nucleoside analogue inhibitor of HBV DNA polymerase in Caucasian children. Data is however limited and it's actual efficacy and safety in children is yet to be determined. We present the case of an 8-year-old Nigerian boy with NS and active hepatitis B virus infection. He went into remission 3 months after commencing oral lamivudine which he had for a year with no significant side effects observed. He remains in remission 3 years later. This, to our knowledge is the first report in literature of successful treatment in an African child. PMID- 21624926 TI - The mismatch negativity: an index of cognitive decline in neuropsychiatric and neurological diseases and in ageing. AB - Cognitive impairment is a core element shared by a large number of different neurological and neuropsychiatric diseases. Irrespective of their different aetiologies and symptomatologies, most appear to converge at the functional deficiency of the auditory-frontal cortex network of auditory discrimination, which indexes cognitive impairment shared by these abnormalities. This auditory frontal cortical deficiency, and hence cognitive decline, can now be objectively measured with the mismatch negativity and its magnetic equivalent. The auditory frontal cortical network involved seems, therefore, to play a pivotal, unifying role in the different abnormalities. It is, however, more likely that the dysfunction that can be detected with the mismatch negativity and its magnetoencephalographic equivalent manifests a more widespread brain disorder, namely, a deficient N-methyl-D-aspartate receptor function, shared by these abnormalities and accounting for most of the cognitive decline. PMID- 21624927 TI - Spatial orienting by left hemisphere language areas: a relict from the past? AB - During evolution, the human brain developed remarkable functional differences between left and right hemispheres. Due to this lateralization, disorders of spatial orienting occur predominantly after right brain damage and disorders of language after left brain damage. In contrast to this general pattern, few individuals show disturbed spatial orienting (spatial neglect) after left brain damage. Using a voxel-based lesion analysis approach, we found that neglect after acute left brain damage is represented in areas typically serving language functions, namely the superior and middle temporal gyri, inferior parietal lobule and insula. Since all except one of these patients also suffered from aphasia, we conclude that lateralization is not just reversed but that both functions (language and spatial orienting) rather are represented in the same left hemisphere regions. We speculate that a representation of spatial orienting in left hemisphere language areas might be a phylogenetic relict in humans, though this representation stays subdominant in the vast majority of individuals. PMID- 21624928 TI - A review of the measurement of grip strength in clinical and epidemiological studies: towards a standardised approach. AB - BACKGROUND: the European Working Group on Sarcopenia in Older People has developed a clinical definition of sarcopenia based on low muscle mass and reduced muscle function (strength or performance). Grip strength is recommended as a good simple measure of muscle strength when 'measured in standard conditions'. However, standard conditions remain to be defined. METHODS: a literature search was conducted to review articles describing the measurement of grip strength listed in Medline, Web of Science and Cochrane Library databases up to 31 December 2009. RESULTS: there is wide variability in the choice of equipment and protocol for measuring grip strength. The Jamar hand dynamometer is the most widely used instrument with established test-retest, inter-rater and intra-rater reliability. However, there is considerable variation in how it is used and studies often provide insufficient information on the protocol followed making comparisons difficult. There is evidence that variation in approach can affect the values recorded. Furthermore, reported summary measures of grip strength vary widely including maximum or mean value, from one, two or three attempts, with either hand or the dominant hand alone. CONCLUSIONS: there is considerable variation in current methods of assessing grip strength which makes comparison between studies difficult. A standardised method would enable more consistent measurement of grip strength and better assessment of sarcopenia. Our approach is described. PMID- 21624930 TI - Perspective: matching, mate choice, and speciation. AB - Matching was developed in the 1960s to match such entities as residents and hospitals, colleges and students, or employers and employees. This approach is based on "preference lists," whereby each participant ranks potential partners according to his/her preferences and tries to match with the highest-ranking partner available. Here, we discuss the implications of matching for the study of mate choice and speciation. Matching differs from classic approaches in several respects, most notably because under this theoretical framework, the formation of mating pairs is context-dependant (i.e., it depends on the configuration of pairings in the entire population), because the stability of mating pairs is considered explicitly, and because mate choice is mutual. The use of matching to study mate choice and speciation is not merely a theoretical curiosity; its application can generate counter-intuitive predictions and lead to conclusions that differ fundamentally from classic theories about sexual selection and speciation. For example, it predicts that when mate choice is mutual and the stability of mating pairs is critical for successful reproduction, sympatric speciation is a robust evolutionary outcome. Yet the application of matching to the study of mate choice and speciation has been largely dominated by theoretical studies. We present the hamlets, a group of brightly colored Caribbean coral reef fishes in the genus Hypoplectrus (Serranidae), as a particularly apt system to test empirically specific predictions generated by the application of matching to mate choice and speciation. PMID- 21624931 TI - Beyond maternal effects in birds: responses of the embryo to the environment. AB - Embryonic growth and development are impacted by environmental conditions. In avian systems, parents tightly control these environments through provisioning of nutrients to the egg and through incubation. Parents can influence embryonic development through egg size, eggshell conductance, hormones, or other substances deposited in eggs and through the onset and temperature of incubation. In addition to these parental influences, evidence suggests that avian embryos are able to perceive and actively respond to their environment during incubation and adjust their own development. Evolution of embryos' responses to developmental environments in birds can be understood in the context of parent-offspring conflicts. When parental investments favor future reproduction over current reproduction, current offspring pay fitness costs, which result in strong selection for offspring that can respond to developmental environments independent of their parents. Here, we review literature indicating that avian embryos actively respond to maternally derived components of the egg, vocalizations, and differences in day length, and we explore these responses in the context of three situations where the consequence of these environments to the fitnesses of offspring and parents differ: the degree of synchrony in hatching, the deposition of hormones in yolks, and seasonal timing of breeding. However, the adaptive significance of responses of embryos to developmental environments arising from parent-offspring conflict has not been adequately explored in birds. PMID- 21624932 TI - Is informal child care associated with childhood obesity? Evidence from Hong Kong's "Children of 1997" birth cohort. AB - BACKGROUND: In Western populations, informal child care is associated with childhood obesity. However, informal child care and obesity share social patterning making evidence from other settings valuable. METHODS: We used multivariable linear and logistic regression models to estimate the association of child care at 6 months and at 3, 5 and 11 years with body mass index (BMI) z score and overweight (including obesity) at 11 years in a Hong Kong Chinese birth cohort. We assessed whether associations varied with sex or socio-economic position (SEP). We used multiple imputation for missing exposures and confounders. RESULTS: Of the original 8327 cohort members, 7933 are alive, participating and living in Hong Kong. At ~11 years, 6796 had their BMI clinically assessed. Higher SEP was associated with informal care. After imputation, informal care at each of 3, 5 or 11 years was separately associated with higher BMI z-score [3 years 0.10, 95% confidence interval (CI) 0.03-0.18, 5 years 0.12, 95% CI 0.04-0.21, 11 years 0.17, 95% CI 0.04-0.31] and with the presence of overweight [odds ratio (OR) 3 years 1.19, 95% CI 1.03-1.37, 5 years OR 1.20, 95% CI 1.03-1.40, 11 years OR 1.21, 95% CI 1.02-1.45], adjusted for sex, SEP and birth weight z-score. Current informal care had the strongest association. However, informal child care at 5 years also contributed. There was no evidence of differences by sex or SEP. CONCLUSIONS: In a developed, non Western setting, informal child care was associated with childhood obesity. Modifiable attributes of informal child care warrant investigation for obesity prevention. PMID- 21624929 TI - Detection of drug resistance mutations at low plasma HIV-1 RNA load in a European multicentre cohort study. AB - BACKGROUND AND OBJECTIVES: Guidelines indicate a plasma HIV-1 RNA load of 500 1000 copies/mL as the minimal threshold for antiretroviral drug resistance testing. Resistance testing at lower viral load levels may be useful to guide timely treatment switches, although data on the clinical utility of this remain limited. We report here the influence of viral load levels on the probability of detecting drug resistance mutations (DRMs) and other mutations by routine genotypic testing in a large multicentre European cohort, with a focus on tests performed at a viral load <1000 copies/mL. METHODS: A total of 16 511 HIV-1 reverse transcriptase and protease sequences from 11 492 treatment-experienced patients were identified, and linked to clinical data on viral load, CD4 T cell counts and antiretroviral treatment history. Test results from 3162 treatment naive patients served as controls. Multivariable analysis was employed to identify predictors of reverse transcriptase and protease DRMs. RESULTS: Overall, 2500/16 511 (15.14%) test results were obtained at a viral load <1000 copies/mL. Individuals with viral load levels of 1000-10000 copies/mL showed the highest probability of drug resistance to any drug class. Independently from other measurable confounders, treatment-experienced patients showed a trend for DRMs and other mutations to decrease at viral load levels <500 copies/mL. CONCLUSIONS: Genotypic testing at low viral load may identify emerging antiretroviral drug resistance at an early stage, and thus might be successfully employed in guiding prompt management strategies that may reduce the accumulation of resistance and cross-resistance, viral adaptive changes, and larger viral load increases. PMID- 21624933 TI - The sequence of cytoreductive nephrectomy: glass half empty or glass half full? PMID- 21624935 TI - Invariant NKT cells are required for airway inflammation induced by environmental antigens. AB - Invariant NKT cells (iNKT cells) are a unique subset of T lymphocytes that rapidly carry out effector functions. In this study, we report that a majority of sterile house dust extracts (HDEs) tested contained antigens capable of activating mouse and human iNKT cells. HDEs had adjuvant-like properties in an ovalbumin (OVA)-induced asthma model, which were dependent on Valpha14i NKT cells, as vaccinated animals deficient for iNKT cells displayed significantly attenuated immune responses and airway inflammation. Furthermore, the administration of HDEs together with OVA mutually augmented the synthesis of cytokines by Valpha14i NKT cells and by conventional CD4(+) T cells in the lung, demonstrating a profound immune response synergy for both Th2 cytokines and IL 17A. These data demonstrate that iNKT cell antigens are far more widely dispersed in the environment than previously anticipated. Furthermore, as the antigenic activity in different houses varied greatly, they further suggest that iNKT cell responses to ambient antigens, particular to certain environments, might promote sensitization to conventional respiratory allergens. PMID- 21624936 TI - Hierarchical organization and early hematopoietic specification of the developing HSC lineage in the AGM region. AB - The aorta-gonad-mesonephros region plays an important role in hematopoietic stem cell (HSC) development during mouse embryogenesis. The vascular endothelial cadherin+ CD45+ (VE-cad+CD45+) population contains the major type of immature pre HSCs capable of developing into long-term repopulating definitive HSCs. In this study, we developed a new coaggregation culture system, which supports maturation of a novel population of CD45-negative (VE-cad+CD45-CD41+) pre-HSCs into definitive HSCs. The appearance of these pre-HSCs precedes development of the VE cad+CD45+ pre-HSCs (termed here type I and type II pre-HSCs, respectively), thus establishing a hierarchical directionality in the developing HSC lineage. By labeling the luminal surface of the dorsal aorta, we show that both type I and type II pre-HSCs are distributed broadly within the endothelial and subendothelial aortic layers, in contrast to mature definitive HSCs which localize to the aortic endothelial layer. In agreement with expression of CD41 in pre-HSCs, in vivo CD41-Cre-mediated genetic tagging occurs in embryonic pre-HSCs and persists in all lymphomyeloid lineages of the adult animal. PMID- 21624937 TI - NKAP is required for T cell maturation and acquisition of functional competency. AB - Newly generated T cells are unable to respond to antigen/MHC. Rather, post selection single-positive thymocytes must undergo T cell maturation to gain functional competency and enter the long-lived naive peripheral T cell pool. This process is poorly understood, as no gene specifically required for T cell maturation has been identified. Here, we demonstrate that loss of the transcriptional repressor NKAP results in a complete block in T cell maturation. In CD4-cre NKAP conditional knockout mice, thymic development including positive selection occurs normally, but there is a cell-intrinsic defect in the peripheral T cell pool. All peripheral naive CD4-cre NKAP conditional knockout T cells were found to be functionally immature recent thymic emigrants. This defect is not simply in cell survival, as the T cell maturation defect was not rescued by a Bcl 2 transgene. Thus, NKAP is required for T cell maturation and the acquisition of functional competency. PMID- 21624938 TI - Hemophagocytosis causes a consumptive anemia of inflammation. AB - Cytopenias of uncertain etiology are commonly observed in patients during severe inflammation. Hemophagocytosis, the histological appearance of blood-eating macrophages, is seen in the disorder hemophagocytic lymphohistiocytosis and other inflammatory contexts. Although it is hypothesized that these phenomena are linked, the mechanisms facilitating acute inflammation-associated cytopenias are unknown. We report that interferon gamma (IFN-gamma) is a critical driver of the acute anemia observed during diverse microbial infections in mice. Furthermore, systemic exposure to physiologically relevant levels of IFN-gamma is sufficient to cause acute cytopenias and hemophagocytosis. Demonstrating the significance of hemophagocytosis, we found that IFN-gamma acts directly on macrophages in vivo to alter endocytosis and provoke blood cell uptake, leading to severe anemia. These findings define a unique pathological process of broad clinical and immunological significance, which we term the consumptive anemia of inflammation. PMID- 21624939 TI - PLZF induces an intravascular surveillance program mediated by long-lived LFA-1 ICAM-1 interactions. AB - Innate-like NKT cells conspicuously accumulate within the liver microvasculature of healthy mice, crawling on the luminal side of endothelial cells, but their general recirculation pattern and the mechanism of their intravascular behavior have not been elucidated. Using parabiotic mice, we demonstrated that, despite their intravascular location, most liver NKT cells failed to recirculate. Antibody blocking experiments established that they were retained locally through constitutive LFA-1-intercellular adhesion molecule (ICAM) 1 interactions. This unprecedented lifelong intravascular residence could be induced in conventional CD4 T cells by the sole expression of promyelocytic leukemia zinc finger (PLZF), a transcription factor specifically expressed in the NKT lineage. These findings reveal the unique genetic and biochemical pathway that underlies the innate intravascular surveillance program of NKT cells. PMID- 21624940 TI - Cigarette smoking, body mass index, and physical fitness changes among male navy personnel. AB - INTRODUCTION: Cigarette smoking has been reported to be higher among deployed military men than among similarly aged civilian or nondeployed men, but the short term effect of smoking on physical fitness among these young healthy men is unclear. This study examined self-reported smoking status and change in objectively measured fitness over 1-4 years while controlling for body mass index (BMI). METHODS: This study included a large sample of male U.S. navy personnel who deployed to Iraq or Kuwait between 2005 and 2008. A mixed modeling procedure was used to determine factors contributing to longitudinal changes in both BMI and fitness (measured by run/walk times, curl-ups, and push-ups). RESULTS: Of the total sample (n = 18,537), the 20% current smokers were more likely than nonsmokers to be enlisted, younger, and have lower BMI measurements at baseline. In addition, smokers had slower 1.5-mile run/walk times and could do fewer curl ups and push-ups compared with nonsmokers. The run/walk time model indicated that over 4 years, smokers (compared with nonsmokers) experienced a significantly greater rate of decrease in cardiorespiratory fitness, even after controlling for changes in BMI. CONCLUSIONS: These results call for continued attention to the problem of nicotine use among young healthy men. PMID- 21624941 TI - Factors predicting recruitment to a UK wide primary care smoking cessation study (the ESCAPE trial). AB - BACKGROUND: Recruiting smokers to smoking cessation trials is challenging and participation rates are often low. Consequently, the interventions evaluated may fail to reach a broad spectrum of the target population, thus compromising the generalizability of the findings. Brief interventions, using proactive recruitment, are likely to attract a broader and more representative proportion of the population. OBJECTIVE: We explored the factors that influenced recruitment into a trial evaluating computer-tailored feedback reports that aimed to help smokers to quit [the ESCAPE (Effectiveness of computer-tailored Smoking Cessation Advice in Primary Care) study] in order to investigate the possibilities for increasing recruitment into smoking cessation trials. METHODS: Current cigarette smokers, identified from GP records, were invited to participate in the study. The main outcome measure was the recruitment rate, i.e. the proportion of participants who responded and were randomized to one of the intervention groups. Predictor variables included geographical region, level of deprivation, practice characteristics and the number and timing of mailings of questionnaires. RESULTS: The recruitment rate varied by practice (2.5-19.8%) and differed significantly between regions (from 16.3% in Scotland to 8.4% in London, P < 0.001). Recruitment decreased significantly by 1.1% between the lowest and highest quintiles of deprivation (P = 0.012), measured by Index of Multiple Deprivation scores, and decreased by 1.33% for every extra 10% smokers identified within a practice population (P = 0.010). Sending reminders increased recruitment by 7.5% (P < 0.001). Multivariable analysis showed region and length of time between mailings were the main predictors of recruitment. CONCLUSIONS: Proactive recruitment methods can increase participation in smoking cessation trials and weighting the target sample in favour of more deprived areas will recruit a more representative sample. The number and timing of mailings to potential participants can also increase recruitment. PMID- 21624942 TI - Spontaneous remission of nephrotic syndrome in membranous nephropathy with chronic renal impairment. AB - BACKGROUND: Spontaneous remission (SR) of nephrotic syndrome, in the absence of immunosuppressive treatment, is relatively common among patients with idiopathic membranous nephropathy (IMN) and normal renal function. However, it has not been reported in patients with chronic renal impairment. METHODS: All patients with IMN who had developed SR in the presence of chronic renal insufficiency were identified among the nephrology departments that belong to the Spanish Group for the Study of Glomerular Diseases (GLOSEN). Their characteristics and outcome after SR were studied. RESULTS: Eleven patients were identified. All of them showed renal insufficiency and nephrotic syndrome at the time of renal biopsy. Serum creatinine (Scr) continued to increase in the following months, reaching a peak value of 2.6 +/- 1.5 mg/dL (range 1.7-6.5). Angiotensin converting enzyme inhibitors or spironolactone were prescribed in 10/11 patients at renal biopsy or shortly after it. Nephrotic proteinuria persisted during the first months of follow-up, but it started to spontaneously decrease 12 +/- 7 months (2-30 months) after renal biopsy. Finally, complete (nine patients) or partial (two patients) remission of nephrotic syndrome was observed. Coinciding with proteinuria remission, renal function tended to improve. Nephrotic syndrome relapsed in two patients, accompanied by a rapid deterioration of renal function. In the remaining nine patients, remission persisted throughout a follow-up of 146 +/- 64 months. Mean Scr at the last visit was 1.9 +/- 0.9 mg/dL and proteinuria 0.2 g/24 h. CONCLUSION: SR of nephrotic syndrome can also be observed in membranous nephropathy patients exhibiting chronic renal impairment. PMID- 21624943 TI - Event-related distress in kidney disease patients. AB - BACKGROUND: Non-dialysis-dependent chronic kidney disease (CKD) and dialysis dependent Stage 5 CKD (CKD5) are associated with a significant physical and psychosocial burden. Little is known, however, about the impact of stressful life events on CKD and CKD5 patients. This study aimed to determine the prevalence of stressful life events in CKD and CKD5 patients and identify the factors correlated with high levels of event-related distress. METHODS: This cross sectional study's sample consisted of 181 patients (91 with non-dialysis dependent CKD Stages 4 and 5, 90 with CKD5) who filled out the Impact of Event Scale (IES), which measures subjective distress related to stressful life events. Other measures included scores from the Medical Outcomes Study Short Form-36, Patient Health Questionnaire-9 (PHQ-9) and Dialysis Symptom Index (DSI). RESULTS: One hundred and three subjects reported stressors on the IES. Almost half the stressors (49.5%) related to personal health; the rest fell into other categories. There were significant differences between the no stressor, low event related distress and high event-related distress groups in age (P < 0.001), PHQ-9 score (P < 0.001) and DSI score (P = 0.002). After adjustment, PHQ-9 score was associated with high event-related distress [odds ratio (OR) 1.20, 95% confidence interval (CI) 1.10-1.32], as was DSI score (OR 1.04, 95% CI 1.02-1.07) in a separate model. CONCLUSIONS: Event-related distress is common in CKD and CKD5 patients. High event-related distress is associated with worse depressive symptoms and greater somatic and emotional symptom burden, even with adjustments for age and gender. The renal practitioner may need to address patients' event related distress in order to provide optimal care. PMID- 21624944 TI - Perspectives on: ion selectivity: design principles for K+ selectivity in membrane transport. PMID- 21624945 TI - Lens intracellular hydrostatic pressure is generated by the circulation of sodium and modulated by gap junction coupling. AB - We recently modeled fluid flow through gap junction channels coupling the pigmented and nonpigmented layers of the ciliary body. The model suggested the channels could transport the secretion of aqueous humor, but flow would be driven by hydrostatic pressure rather than osmosis. The pressure required to drive fluid through a single layer of gap junctions might be just a few mmHg and difficult to measure. In the lens, however, there is a circulation of Na(+) that may be coupled to intracellular fluid flow. Based on this hypothesis, the fluid would cross hundreds of layers of gap junctions, and this might require a large hydrostatic gradient. Therefore, we measured hydrostatic pressure as a function of distance from the center of the lens using an intracellular microelectrode based pressure-sensing system. In wild-type mouse lenses, intracellular pressure varied from ~330 mmHg at the center to zero at the surface. We have several knockout/knock-in mouse models with differing levels of expression of gap junction channels coupling lens fiber cells. Intracellular hydrostatic pressure in lenses from these mouse models varied inversely with the number of channels. When the lens' circulation of Na(+) was either blocked or reduced, intracellular hydrostatic pressure in central fiber cells was either eliminated or reduced proportionally. These data are consistent with our hypotheses: fluid circulates through the lens; the intracellular leg of fluid circulation is through gap junction channels and is driven by hydrostatic pressure; and the fluid flow is generated by membrane transport of sodium. PMID- 21624946 TI - A kinetic analysis of protein transport through the anthrax toxin channel. AB - Anthrax toxin is composed of three proteins: a translocase heptameric channel, (PA(63))(7), formed from protective antigen (PA), which allows the other two proteins, lethal factor (LF) and edema factor (EF), to translocate across a host cell's endosomal membrane, disrupting cellular homeostasis. (PA(63))(7) incorporated into planar phospholipid bilayer membranes forms a channel capable of transporting LF and EF. Protein translocation through the channel can be driven by voltage on a timescale of seconds. A characteristic of the translocation of LF(N), the N-terminal 263 residues of LF, is its S-shaped kinetics. Because all of the translocation experiments reported in the literature have been performed with more than one LF(N) molecule bound to most of the channels, it is not clear whether the S-shaped kinetics are an intrinsic characteristic of translocation kinetics or are merely a consequence of the translocation in tandem of two or three LF(N)s. In this paper, we show both in macroscopic and single-channel experiments that even with only one LF(N) bound to the channel, the translocation kinetics are S shaped. As expected, the translocation rate is slower with more than one LF(N) bound. We also present a simple electrodiffusion model of translocation in which LF(N) is represented as a charged rod that moves subject to both Brownian motion and an applied electric field. The cumulative distribution of first-passage times of the rod past the end of the channel displays S-shaped kinetics with a voltage dependence in agreement with experimental data. PMID- 21624947 TI - An electrostatic potassium channel opener targeting the final voltage sensor transition. AB - Free polyunsaturated fatty acids (PUFAs) modulate the voltage dependence of voltage-gated ion channels. As an important consequence thereof, PUFAs can suppress epileptic seizures and cardiac arrhythmia. However, molecular details for the interaction between PUFA and ion channels are not well understood. In this study, we have localized the site of action for PUFAs on the voltage-gated Shaker K channel by introducing positive charges on the channel surface, which potentiated the PUFA effect. Furthermore, we found that PUFA mainly affects the final voltage sensor movement, which is closely linked to channel opening, and that specific charges at the extracellular end of the voltage sensor are critical for the PUFA effect. Because different voltage-gated K channels have different charge profiles, this implies channel-specific PUFA effects. The identified site and the pharmacological mechanism will potentially be very useful in future drug design of small-molecule compounds specifically targeting neuronal and cardiac excitability. PMID- 21624948 TI - Ion access pathway to the transmembrane pore in P2X receptor channels. AB - P2X receptors are trimeric cation channels that open in response to the binding of adenosine triphosphate (ATP) to a large extracellular domain. The x-ray structure of the P2X4 receptor from zebrafish (zfP2X4) receptor reveals that the extracellular vestibule above the gate opens to the outside through lateral fenestrations, providing a potential pathway for ions to enter and exit the pore. The extracellular region also contains a void at the central axis, providing a second potential pathway. To investigate the energetics of each potential ion permeation pathway, we calculated the electrostatic free energy by solving the Poisson-Boltzmann equation along each of these pathways in the zfP2X4 crystal structure and a homology model of rat P2X2 (rP2X2). We found that the lateral fenestrations are energetically favorable for monovalent cations even in the closed-state structure, whereas the central pathway presents strong electrostatic barriers that would require structural rearrangements to allow for ion accessibility. To probe ion accessibility along these pathways in the rP2X2 receptor, we investigated the modification of introduced Cys residues by methanethiosulfonate (MTS) reagents and constrained structural changes by introducing disulfide bridges. Our results show that MTS reagents can permeate the lateral fenestrations, and that these become larger after ATP binding. Although relatively small MTS reagents can access residues in one of the vestibules within the central pathway, no reactive positions were identified in the upper region of this pathway, and disulfide bridges that constrain movements in that region do not prevent ion conduction. Collectively, these results suggest that ions access the pore using the lateral fenestrations, and that these breathe as the channel opens. The accessibility of ions to one of the chambers in the central pathway likely serves a regulatory function. PMID- 21624949 TI - Ligand-binding domain subregions contributing to bimodal agonism in cyclic nucleotide-gated channels. AB - Cyclic nucleotide-gated (CNG) channels bind cGMP or cAMP in a cytoplasmic ligand binding domain (BD), and this binding typically increases channel open probability (P(o)) without inducing desensitization. However, the catfish CNGA2 (fCNGA2) subtype exhibits bimodal agonism, whereby steady-state P(o) increases with initial cGMP-binding events ("pro" action) up to a maximum of 0.4, but decreases with subsequent cGMP-binding events ("con" action) occurring at concentrations >3 mM. We sought to clarify if low pro-action efficacy was either necessary or sufficient for con action to operate. To find BD residues responsible for con action or low pro-action efficacy or both, we constructed chimeric CNG channels: subregions of the fCNGA2 BD were substituted with corresponding sequence from the rat CNGA4 BD, which does not support con action. Constructs were expressed in frog oocytes and tested by patch clamp of cell-free membranes. For nearly all BD elements, we found at least one construct where replacing that element preserved robust con action, with a ratio of steady-state conductances, g((10 mM cGMP))/g((3 mM cGMP)) < 0.75. When all of the BD sequence C terminal of strand beta6 was replaced, g((10 mM cGMP))/g((3 mM cGMP)) was increased to 0.95 +/- 0.05 (n = 7). However, this apparent attenuation of con action could be explained by an increase in the efficacy of pro action for all agonists, controlled by a conserved "phosphate-binding cassette" motif that contacts ligand; this produces high P(o) values that are less sensitive to shifts in gating equilibrium. In contrast, substituting a single valine in the N terminal helix alphaA abolished con action (g((30 mM cGMP))/g((3 mM cGMP)) increased to 1.26 +/- 0.24; n = 7) without large increases in pro-action efficacy. Our work dissociates the two functional features of low pro-action efficacy and con action, and moreover identifies a separate structural determinant for each. PMID- 21624950 TI - Valerie Weaver: overcoming cancer's stiff resistance. Interview by Caitlin Sedwick. PMID- 21624951 TI - Disturbed flow: p53 SUMOylation in the turnover of endothelial cells. AB - Disturbed blood flow induces apoptosis of vascular endothelial cells, which causes atherosclerosis. In this issue, Heo et al. (2011. J. Cell Biol. doi:10.1083/jcb.201010051) sheds light on p53's role in this phenomenon. Disturbed flow induces peroxynitrite production, which activates protein kinase C zeta and it's binding to the E3 SUMO (small ubiquitin-like modifier) ligase PIASy (protein inhibitor of activated STATy). This leads to p53 SUMOylation and its export to the cytosol, where it binds to the antiapoptotic protein Bcl-2 to induce apoptosis. PMID- 21624952 TI - Real-time imaging of cotranscriptional splicing reveals a kinetic model that reduces noise: implications for alternative splicing regulation. AB - Splicing is a key process that expands the coding capacity of genomes. Its kinetics remain poorly characterized, and the distribution of splicing time caused by the stochasticity of single splicing events is expected to affect regulation efficiency. We conducted a small-scale survey on 40 introns in human cells and observed that most were spliced cotranscriptionally. Consequently, we constructed a reporter system that splices cotranscriptionally and can be monitored in live cells and in real time through the use of MS2-GFP. All small nuclear ribonucleoproteins (snRNPs) are loaded on nascent pre-mRNAs, and spliceostatin A inhibits splicing but not snRNP recruitment. Intron removal occurs in minutes and is best described by a model where several successive steps are rate limiting. Each pre-mRNA molecule is predicted to require a similar time to splice, reducing kinetic noise and improving the regulation of alternative splicing. This model is relevant to other kinetically controlled processes acting on few molecules. PMID- 21624953 TI - Caveolin-1-eNOS signaling promotes p190RhoGAP-A nitration and endothelial permeability. AB - Endothelial barrier function is regulated by adherens junctions (AJs) and caveolae-mediated transcellular pathways. The opening of AJs that is observed in caveolin-1(-/-) (Cav-1(-/-)) endothelium suggests that Cav-1 is necessary for AJ assembly or maintenance. Here, using endothelial cells isolated from Cav-1(-/-) mice, we show that Cav-1 deficiency induced the activation of endothelial nitric oxide synthase (eNOS) and the generation of nitric oxide (NO) and peroxynitrite. We assessed S-nitrosylation and nitration of AJ-associated proteins to identify downstream NO redox signaling targets. We found that the GTPase-activating protein (GAP) p190RhoGAP-A was selectively nitrated at Tyr1105, resulting in impaired GAP activity and RhoA activation. Inhibition of eNOS or RhoA restored AJ integrity and diminished endothelial hyperpermeability in Cav-1(-/-) mice. Thrombin, a mediator of increased endothelial permeability, also induced nitration of p120-catenin-associated p190RhoGAP-A. Thus, eNOS-dependent nitration of p190RhoGAP-A represents a crucial mechanism for AJ disassembly and resultant increased endothelial permeability. PMID- 21624954 TI - RNA polymerase III drives alternative splicing of the potassium channel interacting protein contributing to brain complexity and neurodegeneration. AB - Alternative splicing generates protein isoforms that are conditionally or differentially expressed in specific tissues. The discovery of factors that control alternative splicing might clarify the molecular basis of biological and pathological processes. We found that IL1-alpha-dependent up-regulation of 38A, a small ribonucleic acid (RNA) polymerase III-transcribed RNA, drives the synthesis of an alternatively spliced form of the potassium channel-interacting protein (KCNIP4). The alternative KCNIP4 isoform cannot interact with the gamma-secretase complex, resulting in modification of gamma-secretase activity, amyloid precursor protein processing, and increased secretion of beta-amyloid enriched in the more toxic Abeta x-42 species. Notably, synthesis of the variant KCNIP4 isoform is also detrimental to brain physiology, as it results in the concomitant blockade of the fast kinetics of potassium channels. This alternative splicing shift is observed at high frequency in tissue samples from Alzheimer's disease patients, suggesting that RNA polymerase III cogenes may be upstream determinants of alternative splicing that significantly contribute to homeostasis and pathogenesis in the brain. PMID- 21624955 TI - PKCzeta mediates disturbed flow-induced endothelial apoptosis via p53 SUMOylation. AB - Atherosclerosis is readily observed in regions of blood vessels where disturbed blood flow (d-flow) is known to occur. A positive correlation between protein kinase C zeta (PKCzeta) activation and d-flow has been reported, but the exact role of d-flow-mediated PKCzeta activation in atherosclerosis remains unclear. We tested the hypothesis that PKCzeta activation by d-flow induces endothelial cell (EC) apoptosis by regulating p53. We found that d-flow-mediated peroxynitrite (ONOO(-)) increased PKCzeta activation, which subsequently induced p53 SUMOylation, p53-Bcl-2 binding, and EC apoptosis. Both d-flow and ONOO(-) increased the association of PKCzeta with protein inhibitor of activated STATy (PIASy) via the Siz/PIAS-RING domain (amino acids 301-410) of PIASy, and overexpression of this domain of PIASy disrupted the PKCzeta-PIASy interaction and PKCzeta-mediated p53 SUMOylation. En face confocal microscopy revealed increases in nonnuclear p53 expression, nitrotyrosine staining, and apoptosis in aortic EC located in d-flow areas in wild-type mice, but these effects were significantly decreased in p53(-/-) mice. We propose a novel mechanism for p53 SUMOylation mediated by the PKCzeta-PIASy interaction during d-flow-mediated EC apoptosis, which has potential relevance to early events of atherosclerosis. PMID- 21624956 TI - SH3YL1 regulates dorsal ruffle formation by a novel phosphoinositide-binding domain. AB - Reversible interactions between cytosolic proteins and membrane lipids such as phosphoinositides play important roles in membrane morphogenesis driven by actin polymerization. In this paper, we identify a novel lipid-binding module, which we call the SYLF domain (after the SH3YL1, Ysc84p/Lsb4p, Lsb3p, and plant FYVE proteins that contain it), that is highly conserved from bacteria to mammals. SH3YL1 (SH3 domain containing Ysc84-like 1) strongly bound to phosphatidylinositol 3,4,5-triphosphate (PI(3,4,5)P(3)) and several D5 phosphorylated phosphoinositides through its SYLF domain and was localized to circular dorsal ruffles induced by platelet-derived growth factor stimulation. Interestingly, SHIP2 (the PI(3,4,5)P(3) 5-phosphatase, src-homology 2-containing inositol-5-phosphatase 2) was identified as a binding partner of SH3YL1, and knockdown of these proteins significantly suppressed dorsal ruffle formation. Phosphatidylinositol 3,4-bisphosphate (PI(3,4)P(2)), which is mainly synthesized from PI(3,4,5)P(3) by the action of SHIP2, was enriched in dorsal ruffles, and PI(3,4)P(2) synthesis strongly correlated with formation of the circular membrane structure. These results provide new insight into the molecular mechanism of dorsal ruffle formation and its regulation by phosphoinositide metabolism. PMID- 21624957 TI - Relation of urinary calcium and magnesium excretion to blood pressure: The International Study Of Macro- And Micro-nutrients And Blood Pressure and The International Cooperative Study On Salt, Other Factors, And Blood Pressure. AB - Data indicate an inverse association between dietary calcium and magnesium intakes and blood pressure (BP); however, much less is known about associations between urinary calcium and magnesium excretion and BP in general populations. The authors assessed the relation of BP to 24-hour excretion of calcium and magnesium in 2 cross-sectional studies. The International Study of Macro- and Micro-Nutrients and Blood Pressure (INTERMAP) comprised 4,679 persons aged 40-59 years from 17 population samples in China, Japan, the United Kingdom, and the United States, and the International Cooperative Study on Salt, Other Factors, and Blood Pressure (INTERSALT) comprised 10,067 persons aged 20-59 years from 52 samples around the world. Timed 24-hour urine collections, BP measurements, and nutrient data from four 24-hour dietary recalls (INTERMAP) were collected. In multiple linear regression analyses, urinary calcium excretion was directly associated with BP. After adjustment for multiple confounders (including weight, height, alcohol intake, calcium intake, urinary sodium level, and urinary potassium intake), systolic BP was 1.9 mm Hg higher per each 4.1 mmol per 24 hours (2 standard deviations) of higher urinary calcium excretion (associations were smaller for diastolic BP) in INTERMAP. Qualitatively similar associations were observed in INTERSALT analyses. Associations between magnesium excretion and BP were small and nonsignificant for most of the models examined. The present data suggest that altered calcium homoeostasis, as exhibited by increased calcium excretion, is associated with higher BP levels. PMID- 21624958 TI - The Life Journeys Of Young Women Project: objectives, design, and recruitment results. AB - The Life Journeys of Young Women Project is the first population-based study to examine the role of economic uncertainty throughout early adulthood on age at first childbirth. A retrospective cross-sectional component was added to an existing cohort study that is based on a birth cohort of women born during 1973 1975 in Adelaide, South Australia (n ~ 1,000). An event history calendar instrument was used to obtain data regarding a range of life domains including partnering, educational attainment, home ownership, higher education debt, employment, and pregnancies over a 20-year period (sometimes as detailed as at monthly intervals). Interviews were conducted between 2007 and 2009. An analysis framework applying time-varying and time-constant survival analysis techniques within a life-course framework was developed that will guide analyses to examine the role of duration and life-course timing of economic uncertainty on age at first childbirth. This paper discusses study objectives and design, fieldwork procedures, planned statistical analyses, and recruitment outcomes, focusing on novel features that would facilitate analogous epidemiologic research. PMID- 21624959 TI - Increased risk of cataract among 28,000 patients with celiac disease. AB - Vitamin deficiencies are prevalent in celiac disease (CD) and are associated with cataract formation, but it is unknown whether persons with CD are at increased risk of cataract. The authors' objective in this population-based cohort study was to determine the risk of cataract among persons with biopsy-verified CD. Data on CD were collected from reports on small intestinal biopsies performed between July 1969 and February 2008 in the 28 regional pathology departments in Sweden. The authors identified 28,756 persons with CD (villous atrophy, Marsh pathology stage 3). For each person with CD, Statistics Sweden selected up to 5 controls matched for age and sex from the Total Population Register. Data on cataract were obtained from the Swedish National Hospital Discharge Register and the National Day-Surgery Register. Cox regression analysis was used to estimate the risk of cataract. During a median follow-up period of 9 years, the authors identified 1,159 cataracts among persons with CD (909 were expected) (hazard ratio = 1.28, 95% confidence interval: 1.19, 1.36). The absolute risk of cataract was 397/100,000 person-years in CD, with an excess risk of 86/100,000 person-years. In conclusion, this study found an increased risk of developing cataract in patients with CD. PMID- 21624960 TI - Predicted changes in vegetation structure affect the susceptibility to invasion of bryophyte-dominated subarctic heath. AB - BACKGROUND AND AIMS: A meta-analysis of global change experiments in arctic tundra sites suggests that plant productivity and the cover of shrubs, grasses and dead plant material (i.e. litter) will increase and the cover of bryophytes will decrease in response to higher air temperatures. However, little is known about which effects these changes in vegetation structure will have on seedling recruitment of species and invasibility of arctic ecosystems. METHODS: A field experiment was done in a bryophyte-dominated, species-rich subarctic heath by manipulating the cover of bryophytes and litter in a factorial design. Three phases of seedling recruitment (seedling emergence, summer seedling survival, first-year recruitment) of the grass Anthoxanthum alpinum and the shrub Betula nana were analysed after they were sown into the experimental plots. KEY RESULTS: Bryophyte and litter removal significantly increased seedling emergence of both species but the effects of manipulations of vegetation structure varied strongly for the later phases of recruitment. Summer survival and first-year recruitment were significantly higher in Anthoxanthum. Although bryophyte removal generally increased summer survival and recruitment, seedlings of Betula showed high mortality in early August on plots where bryophytes had been removed. CONCLUSIONS: Large species-specific variation and significant effects of experimental manipulations on seedling recruitment suggest that changes in vegetation structure as a consequence of global warming will affect the abundance of grasses and shrubs, the species composition and the susceptibility to invasion of subarctic heath vegetation. PMID- 21624961 TI - Screening for type 2 diabetes and dysglycemia. AB - Type 2 diabetes mellitus (T2DM) and dysglycemia (impaired glucose tolerance and/or impaired fasting glucose) are increasingly contributing to the global burden of diseases. The authors reviewed the published literature to critically evaluate the evidence on screening for both conditions and to identify the gaps in current understanding. Acceptable, relatively simple, and accurate tools can be used to screen for both T2DM and dysglycemia. Lifestyle modification and/or medication (e.g., metformin) are cost-effective in reducing the incidence of T2DM. However, their application is not yet routine practice. It is unclear whether diabetes-prevention strategies, which influence cardiovascular risk favorably, will also prevent diabetic vascular complications. Cardioprotective therapies, which are cost-effective in preventing complications in conventionally diagnosed T2DM, can be used in screen-detected diabetes, but the magnitude of their effects is unknown. Economic modeling suggests that screening for both T2DM and dysglycemia may be cost-effective, although empirical data on tangible benefits in preventing complications or death are lacking. Screening for T2DM is psychologically unharmful, but the specific impact of attributing the label of dysglycemia remains uncertain. Addressing these gaps will inform the development of a screening policy for T2DM and dysglycemia within a holistic diabetes prevention and control framework combining secondary and high-risk primary prevention strategies. PMID- 21624962 TI - Measuring mortality reductions in cancer screening trials. AB - Randomized trials involving large numbers of people and long follow-up have helped measure the mortality reductions achievable by screening for cancer. However, in many of these trials, the reported reductions have been modest. Part of the reason is the inappropriate way the reductions have been calculated. Analyses have largely ignored the fact that there is a time window in the first several years after screening begins in which there cannot be a sizable mortality reduction, followed by one in which the reductions become evident, and-unless screening is continued-a third window in which mortality rates in the screened group revert to those in the unscreened group. This review uses time-specific mortality ratios to address the timing and extent of the reductions achieved in trials of screening for prostate, breast, and colorectal cancer. The author finds that the mortality reductions reported in the literature have substantially underestimated what might be accomplished with continued screening. The natural history of the disease, the frequency of screening, and the duration of follow-up determine the time patterns in the reductions observed in trials. Without appropriate analyses, results from cancer screening trials will be distorted. PMID- 21624963 TI - Predisease: when does it make sense? AB - Screening often leads to finding conditions that are not at the stage or level that would classify them as disease but, at the same time, are not at a stage or level at which people can be declared entirely disease free. These "in-between" states have sometimes been designated as "predisease." Examples include precancerous lesions, increased intraocular pressure ("preglaucoma"), prediabetes, and prehypertension. When the goal of preventing adverse health outcomes is kept in mind, this review poses the idea that "predisease" as a category on which to act makes sense only if the following 3 conditions are met. First, the people designated as having predisease must be far more likely to develop disease than those not so designated. Second, there must be a feasible intervention that, when targeted to people with predisease, effectively reduces the likelihood of developing disease. Third, the benefits of intervening on predisease must outweigh the harms in the population. A systematic review of screening guidelines (published in 2003-2010) for 4 sample conditions (cervical cancer, glaucoma, diabetes, and hypertension) is included to assess whether they address these issues, followed by a discussion of the framework questions as they pertain to each condition. PMID- 21624964 TI - Optimizing intravenous drug administration by applying pharmacokinetic/pharmacodynamic concepts. AB - This review discusses the ways in which anaesthetists can optimize anaesthetic analgesic drug administration by utilizing pharmacokinetic and pharmacodynamic information. We therefore focus on the dose-response relationship and the interactions between i.v. hypnotics and opioids. For i.v. hypnotics and opioids, models that accurately predict the time course of drug disposition and effect can be applied. Various commercial or experimental drug effect measures have been developed and can be implemented to further fine-tune individual patient-drug titration. The development of advisory and closed-loop feedback systems, which combine and integrate all sources of pharmacological and effect monitoring, has taken the existing kinetic-based administration technology forwards closer to total coverage of the dose-response relationship. PMID- 21624965 TI - Malignant hyperthermia: pharmacology of triggering. AB - Over the past 50 yr, many drugs have been implicated as triggers of malignant hyperthermia (MH), a potentially fatal pharmacogenetic disorder of skeletal muscle calcium regulation. This review discusses the potent inhalation agents as the principal triggers and evidence that the modern agents, desflurane, sevoflurane, and isoflurane, can cause florid MH reactions in the same way as halothane but also are associated with reactions whose onset is delayed for several hours into anaesthesia. There is evidence that the triggering of MH by drugs is dose-dependent but the minimum dose that will trigger the condition is unknown. This has implications for the preparation of anaesthetic machines when used for known or suspected MH patients. While succinylcholine enhances the response of potent inhalation anaesthetics, its role as an inherent trigger of the condition is controversial. Non-depolarizing neuromuscular blocking drugs appear to protect against the development of MH and this may be by blocking excitation-coupled calcium entry-a recently described route of skeletal muscle calcium entry that may also explain the mechanism of the effect of succinylcholine in MH. Another mechanism for extracellular calcium influx, store operated calcium entry, is activated in MH muscle and may explain how a triggered reaction is sustained. Finally, reports of drugs that have been implicated as additional triggers of MH over the past 10 yr are discussed. PMID- 21624966 TI - An update on analgesics. AB - Recent introduction of new analgesics into the clinic is best described as a slow process with activity classified into two main areas: improving analgesic efficacy/potency and reducing side-effect profile. This review article describes some of the recent advances with an emphasis on use in the acute setting. In this respect, opioids continue to be the mainstay (but not the only) analgesic and there have been important improvements in their clinical effect profile. For example, tapentadol has been introduced as a mixed opioid and norepinephrine uptake inhibitor which, unlike tramadol, does not require metabolic activation and does not suffer from isomer-dependent pharmacodynamics. Opioid antagonists have received much attention recently either used alone, methylnaltrexone (s.c) or alvimopan (p.o), or in combination, Targinact (oxycodone/naloxone), and appear to be effective in reducing opioid side-effects such as those in the gastrointestinal tract. Other agents where there has been recent development include the use of gabapentin, methylxanthines, and local anaesthetics. An interesting area of translation of basic research is in the inhibition of breakdown of endogenous opioids with opiorphin, targeting of the endocannabinoid system, and the use of ampakines to obtund opioid-induced side-effects. It is clear that there is still much work to be done, but the need for highly efficacious analgesics with good side-effect profile remains. PMID- 21624967 TI - The role of hyperpolarization-activated cationic current in spike-time precision and intrinsic resonance in cortical neurons in vitro. AB - Hyperpolarization-activated cyclic nucleotide modulated current (I(h)) sets resonance frequency within the theta-range (5-12 Hz) in pyramidal neurons. However, its precise contribution to the temporal fidelity of spike generation in response to stimulation of excitatory or inhibitory synapses remains unclear. In conditions where pharmacological blockade of I(h) does not affect synaptic transmission, we show that postsynaptic h-channels improve spike time precision in CA1 pyramidal neurons through two main mechanisms. I(h) enhances precision of excitatory postsynaptic potential (EPSP)--spike coupling because I(h) reduces peak EPSP duration. I(h) improves the precision of rebound spiking following inhibitory postsynaptic potentials (IPSPs) in CA1 pyramidal neurons and sets pacemaker activity in stratum oriens interneurons because I(h) accelerates the decay of both IPSPs and after-hyperpolarizing potentials (AHPs). The contribution of h-channels to intrinsic resonance and EPSP waveform was comparatively much smaller in CA3 pyramidal neurons. Our results indicate that the elementary mechanisms by which postsynaptic h-channels control fidelity of spike timing at the scale of individual neurons may account for the decreased theta-activity observed in hippocampal and neocortical networks when h-channel activity is pharmacologically reduced. PMID- 21624968 TI - Augmented skeletal muscle hyperaemia during hypoxic exercise in humans is blunted by combined inhibition of nitric oxide and vasodilating prostaglandins. AB - Exercise hyperaemia in hypoxia is augmented relative to the same level of exercise in normoxia. At moderate exercise intensities, the mechanism(s) underlying this augmented response are currently unclear. We tested the hypothesis that endothelium-derived nitric oxide (NO) and vasodilating prostaglandins (PGs) contribute to the augmented muscle blood flow during hypoxic exercise relative to normoxia. In 10 young healthy adults, we measured forearm blood flow (FBF; Doppler ultrasound) and calculated the vascular conductance (FVC) responses during 5 min of rhythmic handgrip exercise at 20% maximal voluntary contraction in normoxia (NormEx) and isocapnic hypoxia (HypEx; O2 saturation ~85%) before and after local intra-brachial combined blockade of NO synthase (NOS; via N(G)-monomethyl-L-arginine: L-NMMA) and cyclooxygenase (COX; via ketorolac). All trials were performed during local alpha- and beta adrenoceptor blockade to eliminate sympathoadrenal influences on vascular tone and thus isolate local vasodilatation. Arterial and deep venous blood gases were measured and oxygen consumption (VO2) was calculated. In control (saline) conditions, FBF after 5 min of exercise in hypoxia was greater than in normoxia (345 +/- 21 ml min(-1) vs. 297 +/- 18 ml min(-1); P < 0.05). After NO-PG block, the compensatory increase in FBF during hypoxic exercise was blunted ~50% and thus was reduced compared with control hypoxic exercise (312 +/- 19 ml min(-1); P < 0.05), but this was not the case in normoxia (289 +/- 15 ml min(-1); P = 0.33). The lower FBF during hypoxic exercise was associated with a compensatory increase in O2 extraction, and thus VO2 was maintained at normal control levels (P = 0.64 0.99). We conclude that under the experimental conditions employed, NO and PGs have little role in normoxic exercise hyperaemia whereas combined NO-PG inhibition reduces hypoxic exercise hyperaemia and abolishes hypoxic vasodilatation at rest. Additionally, VO2 of the tissue was maintained in hypoxic conditions at rest and during exercise, despite attenuated oxygen delivery following NO-PG blockade, due to an increase in O2 extraction at the level of the muscle. PMID- 21624969 TI - Dietary composition programmes placental phenotype in mice. AB - Dietary composition during pregnancy influences fetal and adult phenotype but its effects on placental phenotype remain largely unknown. Using molecular, morphological and functional analyses, placental nutrient transfer capacity was examined in mice fed isocaloric diets containing 23%, 18% or 9% casein (C) during pregnancy. At day 16, placental transfer of glucose, but not methyl aminoisobutyric acid (MeAIB), was greater in C18 and C9 than C23 mice, in association with increased placental expression of the glucose transporter Slc2a1/GLUT1, and the growth factor Igf2. At day 19, placental glucose transport remained high in C9 mice while MeAIB transfer was less in C18 than C23 mice, despite greater placental weights in C18 and C9 than C23 mice. Placental System A amino acid transporter expression correlated with protein intake at day 19. Relative growth of transport verses endocrine zones of the placenta was influenced by diet at both ages without changing the absolute volume of the transport surface. Fetal weight was unaffected by diet at day 16 but was reduced in C9 animals by day 19. Morphological and functional adaptations in placental phenotype, therefore, occur to optimise nutrient transfer when dietary composition is varied, even subtly. This has important implications for the intrauterine programming of life expectancy. PMID- 21624970 TI - Contributions of descending and ascending pathways to corticomuscular coherence in humans. AB - Corticomuscular coherence in the beta frequency band (15-30 Hz) has been demonstrated in both humans and monkeys, but its origin and functional role are still unclear. Phase-frequency plots produced by traditional coherence analysis are often complex. Some subjects show a clear linear phase-frequency relationship (indicative of a fixed delay) but give shorter delays than expected; others show a constant phase across frequencies. Recent evidence suggests that oscillations may be travelling around a peripheral sensorimotor loop. We recorded sensorimotor EEGs and EMGs from three intrinsic hand muscles in human subjects performing a precision grip task, and applied directed coherence (Granger causality) analysis to explore this system. Directed coherence was significant in both descending (EEG -> EMG) and ascending(EMG -> EEG) directions at beta frequencies. Average phase delays of 26.4 ms for the EEG -> EMG direction and 29.5 ms for the EMG -> EEG direction were closer to the expected conduction times for these pathways than the average delays estimated from coherence phase (7.9 ms). Subjects were sub-divided into different groups, based on the sign of the slope of the linear relation between corticomuscular coherence phase and frequency (positive, negative or zero). Analysis separated by these groups suggested that different relative magnitudes of EEG -> EMG and EMG -> EEG directed coherence might underlie the observed inter-individual differences in coherence phase.These results confirm the complex nature of corticomuscular coherence with contributions from both descending and ascending pathways. PMID- 21624971 TI - Oligogenic heterozygosity in individuals with high-functioning autism spectrum disorders. AB - Autism spectrum disorders (ASDs) are a heterogeneous group of neuro-developmental disorders. While significant progress has been made in the identification of genes and copy number variants associated with syndromic autism, little is known to date about the etiology of idiopathic non-syndromic autism. Sanger sequencing of 21 known autism susceptibility genes in 339 individuals with high-functioning, idiopathic ASD revealed de novo mutations in at least one of these genes in 6 of 339 probands (1.8%). Additionally, multiple events of oligogenic heterozygosity were seen, affecting 23 of 339 probands (6.8%). Screening of a control population for novel coding variants in CACNA1C, CDKL5, HOXA1, SHANK3, TSC1, TSC2 and UBE3A by the same sequencing technology revealed that controls were carriers of oligogenic heterozygous events at significantly (P < 0.01) lower rate, suggesting oligogenic heterozygosity as a new potential mechanism in the pathogenesis of ASDs. PMID- 21624973 TI - Molecular mechanisms of MLC1 and GLIALCAM mutations in megalencephalic leukoencephalopathy with subcortical cysts. AB - Megalencephalic leukoencephalopathy with subcortical cysts (MLC) is a rare leukodystrophy caused by mutations in MLC1 or GLIALCAM. The GLIALCAM gene product functions as an MLC1 beta-subunit. We aim to further clarify the molecular mechanisms of MLC caused by mutations in MLC1 or GLIALCAM. For this purpose, we analyzed a human post-mortem brain obtained from an MLC patient, who was homozygous for a missense mutation (S69L) in MLC1. We showed that this mutation affects the stability of MLC1 in vitro and reduces MLC1 protein levels in the brain to almost undetectable. However, the amount of GlialCAM and its localization were nearly unaffected, indicating that MLC1 is not necessary for GlialCAM expression or targeting. These findings were supported by experiments in primary astrocytes and in heterologous cells. In addition, we demonstrated that MLC1 and GlialCAM form homo- and hetero-complexes and that MLC-causing mutations in GLIALCAM mainly reduce the formation of GlialCAM homo-complexes, leading to a defect in the trafficking of GlialCAM alone to cell junctions. GLIALCAM mutations also affect the trafficking of its associated molecule MLC1, explaining why GLIALCAM and MLC1 mutations lead to the same disease: MLC. PMID- 21624972 TI - Pathogenity of some limb girdle muscular dystrophy mutations can result from reduced anchorage to myofibrils and altered stability of calpain 3. AB - Calpain 3 (CAPN3) is a muscle-specific, calcium-dependent proteinase that is mutated in Limb Girdle Muscle Dystrophy type 2A. Most pathogenic missense mutations in LGMD2A affect CAPN3's proteolytic activity; however, two mutations, D705G and R448H, retain activity but nevertheless cause muscular dystrophy. Previously, we showed that D705G and R448H mutations reduce CAPN3s ability to bind to titin in vitro. In this investigation, we tested the consequence of loss of titin binding in vivo and examined whether this loss can be an underlying pathogenic mechanism in LGMD2A. To address this question, we created transgenic mice that express R448H or D705G in muscles, on wild-type (WT) CAPN3 or knock-out background. Both mutants were readily expressed in insect cells, but when D705G was expressed in skeletal muscle, it was not stable enough to study. Moreover, the D705G mutation had a dominant negative effect on endogenous CAPN3 when expressed on a WT background. The R448H protein was stably expressed in muscles; however, it was more rapidly degraded in muscle extracts compared with WT CAPN3. Increased degradation of R448H was due to non-cysteine, cellular proteases acting on the autolytic sites of CAPN3, rather than autolysis. Fractionation experiments revealed a significant decrease of R448H from the myofibrillar fraction, likely due to the mutant's inability to bind titin. Our data suggest that R448H and D705G mutations affect both CAPN3s anchorage to titin and its stability. These studies reveal a novel mechanism by which mutations that spare enzymatic activity can still lead to calpainopathy. PMID- 21624974 TI - Small molecules for big tasks. PMID- 21624975 TI - Different developmental potential of pluripotent stem cells generated by different reprogramming strategies. PMID- 21624976 TI - Reflections on food security under water scarcity. AB - Forecasts on population growth and economic development indicate that there will be substantial increases in food demand for the forthcoming decades. We focus here on the water requirements of food production, on the issue of whether there would be enough water to produce sufficient food in the future, and we offer options to face this challenge based on recent trends observed in some agricultural systems. Given the competition for water faced by the agricultural sector, and the uncertainties associated with climate change, improving the efficiency of water use in both rain-fed and irrigated systems is the main avenue to face the challenge. In rain-fed agriculture, managing the risk associated with rainfall variability is a promising option to increase productivity. In irrigated systems, a case study on the improvements in water productivity in Andalusia, Spain, is used to illustrate some of the opportunities to make progress. Progress in reducing irrigation water use in recent decades has been substantial, but decreasing the consumptive use of crops is a much more difficult challenge. The need for more research and technology transfer on improving water-limited crop production is highlighted, and emphasis is placed on interdisciplinary approaches to gain the insight needed to achieve new breakthroughs that would help in tackling this complex problem. PMID- 21624977 TI - Hydrogen sulphide enhances photosynthesis through promoting chloroplast biogenesis, photosynthetic enzyme expression, and thiol redox modification in Spinacia oleracea seedlings. AB - Hydrogen sulphide (H(2)S) is emerging as a potential messenger molecule involved in modulation of physiological processes in animals and plants. In this report, the role of H(2)S in modulating photosynthesis of Spinacia oleracea seedlings was investigated. The main results are as follows. (i) NaHS, a donor of H(2)S, was found to increase the chlorophyll content in leaves. (ii) Seedlings treated with different concentrations of NaHS for 30 d exhibited a significant increase in seedling growth, soluble protein content, and photosynthesis in a dose-dependent manner, with 100 MUM NaHS being the optimal concentration. (iii) The number of grana lamellae stacking into the functional chloroplasts was also markedly increased by treatment with the optimal NaHS concentration. (iv) The light saturation point (Lsp), maximum net photosynthetic rate (Pmax), carboxylation efficiency (CE), and maximal photochemical efficiency of photosystem II (F(v)/F(m)) reached their maximal values, whereas the light compensation point (Lcp) and dark respiration (Rd) decreased significantly under the optimal NaHS concentration. (v) The activity of ribulose-1,5-bisphosphate carboxylase (RuBISCO) and the protein expression of the RuBISCO large subunit (RuBISCO LSU) were also significantly enhanced by NaHS. (vi) The total thiol content, glutathione and cysteine levels, internal concentration of H(2)S, and O acetylserine(thiol)lyase and L-cysteine desulphydrase activities were increased to some extent, suggesting that NaHS also induced the activity of thiol redox modification. (vii) Further studies using quantitative real-time PCR showed that the gene encoding the RuBISCO large subunit (RBCL), small subunit (RBCS), ferredoxin thioredoxin reductase (FTR), ferredoxin (FRX), thioredoxin m (TRX-m), thioredoxin f (TRX-f), NADP-malate dehydrogenase (NADP-MDH), and O acetylserine(thiol)lyase (OAS) were up-regulated, but genes encoding serine acetyltransferase (SERAT), glycolate oxidase (GYX), and cytochrome oxidase (CCO) were down-regulated after exposure to the optimal concentration of H(2)S. These findings suggest that increases in RuBISCO activity and the function of thiol redox modification may underlie the amelioration of photosynthesis and that H(2)S plays an important role in plant photosynthesis regulation by modulating the expression of genes involved in photosynthesis and thiol redox modification. PMID- 21624978 TI - Allelic interaction of F1 pollen sterility loci and abnormal chromosome behaviour caused pollen sterility in intersubspecific autotetraploid rice hybrids. AB - The intersubspecific hybrids of autotetraploid rice has many features that increase rice yield, but lower seed set is a major hindrance in its utilization. Pollen sterility is one of the most important factors which cause intersubspecific hybrid sterility. The hybrids with greater variation in seed set were used to study how the F(1) pollen sterile loci (S-a, S-b, and S-c) interact with each other and how abnormal chromosome behaviour and allelic interaction of F(1) sterility loci affect pollen fertility and seed set of intersubspecific autotetraploid rice hybrids. The results showed that interaction between pollen sterility loci have significant effects on the pollen fertility of autotetraploid hybrids, and pollen fertility further decreased with an increase in the allelic interaction of F(1) pollen sterility loci. Abnormal ultra-structure and microtubule distribution patterns during pollen mother cell (PMC) meiosis were found in the hybrids with low pollen fertility in interphase and leptotene, suggesting that the effect-time of pollen sterility loci interaction was very early. There were highly significant differences in the number of quadrivalents and bivalents, and in chromosome configuration among all the hybrids, and quadrivalents decreased with an increase in the seed set of autotetraploid hybrids. Many different kinds of chromosomal abnormalities, such as chromosome straggling, chromosome lagging, asynchrony of chromosome disjunction, and tri fission were found during the various developmental stages of PMC meiosis. All these abnormalities were significantly higher in sterile hybrids than in fertile hybrids, suggesting that pollen sterility gene interactions tend to increase the chromosomal abnormalities which cause the partial abortion of male gametes and leads to the decline in the seed set of the autotetraploid rice hybrids. PMID- 21624979 TI - Integrated functions among multiple starch synthases determine both amylopectin chain length and branch linkage location in Arabidopsis leaf starch. AB - This study assessed the impact on starch metabolism in Arabidopsis leaves of simultaneously eliminating multiple soluble starch synthases (SS) from among SS1, SS2, and SS3. Double mutant ss1- ss2- or ss1- ss3- lines were generated using confirmed null mutations. These were compared to the wild type, each single mutant, and ss1- ss2- ss3- triple mutant lines grown in standardized environments. Double mutant plants developed similarly to the wild type, although they accumulated less leaf starch in both short-day and long-day diurnal cycles. Despite the reduced levels in the double mutants, lines containing only SS2 and SS4, or SS3 and SS4, are able to produce substantial amounts of starch granules. In both double mutants the residual starch was structurally modified including higher ratios of amylose:amylopectin, altered glucan chain length distribution within amylopectin, abnormal granule morphology, and altered placement of alpha(1 >6) branch linkages relative to the reducing end of each linear chain. The data demonstrate that SS activity affects not only chain elongation but also the net result of branch placement accomplished by the balanced activities of starch branching enzymes and starch debranching enzymes. SS3 was shown partially to overlap in function with SS1 for the generation of short glucan chains within amylopectin. Compensatory functions that, in some instances, allow continued residual starch production in the absence of specific SS classes were identified, probaby accomplished by the granule bound starch synthase GBSS1. PMID- 21624980 TI - Overexpression of EVE1, a novel ubiquitin family protein, arrests inflorescence stem development in Arabidopsis. AB - In Arabidopsis, inflorescence stem formation is a critical process in phase transition from the vegetative to the reproductive state. Although inflorescence stem development has been reported to depend on the expression of a variety of genes during floral induction and repression, little is known about the molecular mechanisms involved in the control of inflorescence stem formation. By activation T-DNA tagging mutagenesis of Arabidopsis, a dominant gain-of-function mutation, eve1-D (eternally vegetative phase1-Dominant), which has lost the ability to form an inflorescence stem, was isolated. The eve1-D mutation exhibited a dome-shaped primary shoot apical meristem (SAM) in the early vegetative stage, similar to that seen in the wild-type SAM. However, the SAM in the eve1-D mutation failed to transition into an inflorescence meristem (IM) and eventually reached senescence without ever leaving the vegetative phase. The eve1-D mutation also displayed pleiotropic phenotypes, including lobed and wavy rosette leaves, short petioles, and an increased number of rosette leaves. Genetic analysis indicated that the genomic location of the EVE1 gene in Arabidopsis thaliana corresponded to a bacterial artificial chromosome (BAC) F4C21 from chromosome IV at ~17cM which encoded a novel ubiquitin family protein (At4g03350), consisting of a single exon. The EVE1 protein is composed of 263 amino acids, contains a 52 amino acid ubiquitin domain, and has no glycine residue related to ubiquitin activity at the C-terminus. The eve1-D mutation provides a way to study the regulatory mechanisms that control phase transition from the vegetative to the reproductive state. PMID- 21624981 TI - Clinical reasoning: A young adult presents with focal weakness and hemorrhagic brain lesions. PMID- 21624982 TI - Teaching NeuroImages: Unilateral prosopometamorphopsia as a dominant hemisphere specific disconnection sign. PMID- 21624983 TI - Patient page. The influence of lacunes on cognitive function. PMID- 21624985 TI - Stress and the risk of multiple sclerosis. AB - OBJECTIVE: Several studies have shown that stressful life events are associated with a subsequent significant increase in risk of multiple sclerosis (MS) exacerbations. We wanted to study prospectively whether stress can increase the risk of developing the disease itself. METHODS: We studied 2 cohorts of female nurses: the Nurses' Health Study (NHS) (n = 121,700) followed from 1976 and the Nurses' Health Study II (NHS II) (n = 116,671) followed from 1989. The risk of MS after self-report on general stress at home and at work in the NHS in 1982 was studied prospectively using Cox regression. Logistic regression was used to retrospectively estimate the effects of physical and sexual abuse in childhood and adolescence collected in the NHS II 2001. We identified 77 cases of MS in the NHS by 2005 and 292 in the NHS II by 2004. All analyses were adjusted for age, ethnicity, latitude of birth, body mass index at age 18, and smoking. RESULTS: We found no increased risk of MS associated with severe stress at home in the NHS (hazard ratio 0.85 [95% confidence interval (CI)] 0.32-2.26). No significantly increased risk of MS was found among those who reported severe physical abuse during childhood (odds ratio [OR] 0.68, 95% CI 0.41-1.14) or adolescence (OR 0.77, 95% CI 0.46-1.28) or those having been repeatedly forced into sexual activity in childhood (OR 1.47, 95% CI 0.87-2.48) or adolescence (OR 1.21, 95% CI 0.68-2.17). CONCLUSIONS: These results do not support a major role of stress in the development of the disease, but repeated and more focused measures of stress are needed to firmly exclude stress as a potential risk factor for MS. PMID- 21624986 TI - Predicting survival in frontotemporal dementia with motor neuron disease. AB - OBJECTIVE: To determine whether clinical and demographic features are associated with prognosis in patients with frontotemporal dementia and motor neuron disease (FTD-MND). METHODS: This was a case series of FTD-MND categorized according to behavioral- or language-dominant symptoms at presentation and throughout the disease course. Demographic, clinical, imaging, and survival data were analyzed with respect to dominant FTD-MND type. Voxel-based morphometry was used to assess and compare regional patterns of atrophy in behavioral- and language-dominant FTD MND types. RESULTS: Of the 56 patients with FTD-MND who were identified, 31 had dominant behavioral symptoms and 25 had dominant language symptoms; 53 patients had died. A survival difference was present between types, with patients with behavioral-dominant symptoms surviving 506 days longer than patients with language-dominant symptoms (mean 1,397 vs 891 days; p = 0.002). There was also a difference in time from diagnosis to death (p = 0.02) between groups. Patients with language-dominant disease were more likely to have bulbar-onset than limb onset motor neuron disease (MND) (p = 0.01). There was a similar pattern of frontal and temporal lobe atrophy in both types, although there was some evidence for the behavioral type to have more frontal atrophy and the language type to have more left temporal atrophy. CONCLUSIONS: In our series of patients with FTD MND, language-dominant FTD-MND was associated with bulbar-onset MND and a shorter survival. There was also evidence that the dominant FTD-MND type is related to differences in brain atrophy patterns. PMID- 21624988 TI - Real-life driving outcomes in Parkinson disease. AB - OBJECTIVE: To determine the incidence of and risk factors for driving outcomes in drivers with Parkinson disease (PD). METHODS: In a prospective cohort study, we ascertained the time until driving cessation, a crash, or a traffic citation using self-report and state Department of Transportation records in 106 licensed, active drivers with PD and 130 controls. RESULTS: Drivers with PD stopped driving earlier than controls, hazard ratio (95% confidence interval) = 7.09 (3.66 13.75), p < 0.001. Cumulative incidence of driving cessation at 2 years after baseline was 17.6% (11.5%-26.5%) for PD and 3.1% (1.2%-8.1%) for controls. No significant differences between groups on times to first crash or citation were detected. However, the number of observed crashes was low. Cox proportional hazards models showed that significant baseline risk factors for driving cessation in PD were older age, preference to be driven by somebody else, positive crash history, use of compensatory strategies, low driving exposure, impairments in visual perception (especially visual processing speed and attention) and cognitive abilities, parkinsonism (especially activities of daily living score and total daily dose of antiparkinsonian medications), and higher error counts on a road test. Within PD, crashes were associated with poorer postural stability and history of driving citations, and citations were associated with younger age and road errors at baseline. CONCLUSIONS: Drivers with PD are at a higher risk of driving cessation than elderly control drivers. A battery evaluating motor and nonmotor aspects of PD, driving record, and performance can be useful in assessing future driving outcomes in PD. PMID- 21624989 TI - Sensory neuropathy as part of the cerebellar ataxia neuropathy vestibular areflexia syndrome. AB - OBJECTIVE: The syndrome of cerebellar ataxia with bilateral vestibulopathy was delineated in 2004. Sensory neuropathy was mentioned in 3 of the 4 patients described. We aimed to characterize and estimate the frequency of neuropathy in this condition, and determine its typical MRI features. METHODS: Retrospective review of 18 subjects (including 4 from the original description) who met the criteria for bilateral vestibulopathy with cerebellar ataxia. RESULTS: The reported age at onset range was 39-71 years, and symptom duration was 3-38 years. The syndrome was identified in one sibling pair, suggesting that this may be a late-onset recessive disorder, although the other 16 cases were apparently sporadic. All 18 had sensory neuropathy with absent sensory nerve action potentials, although this was not apparent clinically in 2, and the presence of neuropathy was not a selection criterion. In 5, the loss of pinprick sensation was virtually global, mimicking a neuronopathy. However, findings in the other 11 with clinically manifest neuropathy suggested a length-dependent neuropathy. MRI scans showed cerebellar atrophy in 16, involving anterior and dorsal vermis, and hemispheric crus I, while 2 were normal. The inferior vermis and brainstem were spared. CONCLUSIONS: Sensory neuropathy is an integral component of this syndrome. It may result in severe sensory loss, which contributes significantly to the disability. The MRI changes are nonspecific, but, coupled with loss of sensory nerve action potentials, may aid diagnosis. We propose a new name for the condition: cerebellar ataxia with neuropathy and bilateral vestibular areflexia syndrome (CANVAS). PMID- 21624990 TI - Clinical and MRI characteristics of acute migrainous infarction. AB - OBJECTIVE: Migrainous infarction is considered a rare complication of migraine. Although several studies reported silent brain lesions on neuroimaging in patients with migraine with aura, knowledge about lesion patterns in acute migrainous infarction is scarce. We investigated clinical and MRI characteristics in a series of patients with migraine-associated acute cerebral ischemia. METHODS: Seventeen patients among 8,137 stroke patients over an 11-year period were included. All had undergone a dedicated stroke workup including diffusion weighted imaging (DWI) and a detailed assessment of clinical features and of vascular risk factors. RESULTS: The majority of patients presented with prolonged aura symptoms (visual aura 82.3%, sensory dysfunction 41.2%, and aphasia 5.9%; median NIH Stroke Scale score 2). Presentation at hospital was significantly delayed after symptom onset (mean 33 hours). A total of 70.6% had acute ischemic lesions in the posterior circulation; the middle cerebral artery territory was affected in 29.4%. Small lesions were present in 64.7%; multiple lesions were found in 41.2%. No overlapping ischemic lesions of different vascular territories were found. The prevalence of a patent foramen ovale was high (64.7%). CONCLUSIONS: This study supports previous observations that migrainous infarction mostly occurs in the posterior circulation, and in younger women with a history of migraine with aura. Acute ischemic lesions were often multiple and located in distinct arterial territories. As there were no overlapping ischemic lesions, hemodynamic compromise during the development of migraine is unlikely the cause of infarction. Differentiation between migrainous infarction and prolonged migraine aura is difficult and associated with delayed admission of patients. PMID- 21624991 TI - Potential utility of conventional MRI signs in diagnosing pseudoprogression in glioblastoma. AB - OBJECTIVE: To examine the potential utility of conventional MRI signs in differentiating pseudoprogression (PsP) from early progression (EP). METHODS: This retrospective study reviewed initial postradiotherapy MRI scans of 321 patients with glioblastoma undergoing chemotherapy and radiotherapy. A total of 93 patients were found to have new or increased enhancing mass lesions, raising the possibility of PsP. Final diagnosis of PsP or EP was established upon review of surgical specimens from a second resection or by clinical and radiologic follow-up. A total of 11 MRI signs potentially helpful in the differentiation between PsP and EP were examined on the initial post-RT MRI and were correlated with the final diagnosis through chi(2) or Fisher exact test. RESULTS: Sixty three (67.7%) of the 93 patients had EP, of which 22 (34.9%) were diagnosed by pathology. Thirty patients (32.3%) had PsP; 6 (16.7% of the 30) were diagnosed by pathology. Subependymal enhancement was predictive for EP (p = 0.001) with 38.1% sensitivity, 93.3% specificity, and 41.8% negative predictive value. The other 10 signs had no predictive value (p = 0.06-1.0). CONCLUSIONS: Conventional MRI signs have limited utility in diagnosing PsP in patients with recently treated glioblastomas and worsening enhancing lesions. We did not find a sign with a high negative predictive value for PsP that would have been the most useful for the clinical physician. When present, subependymal spread of the enhancing lesion is a useful MRI marker in identifying EP rather than PsP. PMID- 21624992 TI - Prognostic importance of serial postoperative EEGs after anterior temporal lobectomy. AB - OBJECTIVE: To assess the value of postoperative EEG in predicting seizure outcome and seizure recurrence following antiepileptic drug (AED) withdrawal in patients with mesial temporal lobe epilepsy with hippocampal sclerosis (MTLE-HS). METHODS: We studied 262 consecutive patients with MTLE-HS with serial EEGs at 3 months, and at 1, 2, and 3 years after anterior temporal lobectomy (ATL), and considered the presence of interictal epileptiform discharges (IED) as abnormal. We attempted AED withdrawal in all seizure-free patients. We defined favorable outcome as freedom from seizures/auras during the entire follow-up period (outcome 1) and during terminal 1-year follow-up (outcome 2). RESULTS: During mean follow-up period of 7.6 (range 5-12) years, 129 (49.2%) patients had favorable outcome 1 and 218 (83.2%) had favorable outcome 2. Of 225 (85.9%) patients in whom AED withdrawal was attempted, 61 (27.1%) had seizure recurrence. Compared to patients with normal EEG, those with IED on 1-year post-ATL EEG had a 3-fold increased risk for unfavorable outcome 1 and 7-fold increased risk for unfavorable outcome 2. The patients in whom all the 4 EEGs were abnormal had 9 fold odds for unfavorable outcome 1 and 26-fold odds for unfavorable outcome 2. An abnormal EEG at 1 year increased the risk of seizure recurrence following AED withdrawal by 2.6-fold. CONCLUSIONS: Post-ATL EEG predicts seizure outcome and seizure recurrence following AED withdrawal. Serial EEGs predict outcome better than single EEG. This information will be helpful in counseling of patients after ATL, and in making rational decisions on AED withdrawal. PMID- 21624993 TI - Diabetes insipidus as a first manifestation in multiple sclerosis. PMID- 21624994 TI - JC virus granule cell neuronopathy and hyper-IgE in HIV disease. PMID- 21624995 TI - Giant thoracic meningocele associated with neurofibromatosis 1. PMID- 21624996 TI - The MoCA: well-suited screen for cognitive impairment in Parkinson disease. PMID- 21624997 TI - Local selection across a latitudinal gradient shapes nucleotide diversity in balsam poplar, Populus balsamifera L. AB - Molecular studies of adaptive evolution often focus on detecting selective sweeps driven by positive selection on a species-wide scale; however, much adaptation is local, particularly of ecologically important traits. Here, we look for evidence of range-wide and local adaptation at candidate genes for adaptive phenology in balsam poplar, Populus balsamifera, a widespread forest tree whose range extends across environmental gradients of photoperiod and growing season length. We examined nucleotide diversity of 27 poplar homologs of the flowering-time network a group of genes that control plant developmental phenology through interactions with environmental cues such as photoperiod and temperature. Only one gene, ZTL2, showed evidence of reduced diversity and an excess of fixed replacement sites, consistent with a species-wide selective sweep. Two other genes, LFY and FRI, harbored high levels of nucleotide diversity and exhibited elevated differentiation between northern and southern accessions, suggesting local adaptation along a latitudinal gradient. Interestingly, FRI has also been identified as a target of local selection between northern and southern accessions of Arabidopsis thaliana, indicating that this gene may be commonly involved in ecological adaptation in distantly related species. Our findings suggest an important role for local selection shaping molecular diversity and reveal limitations of inferring molecular adaptation from analyses designed only to detect species-wide selective sweeps. PMID- 21624998 TI - Regulation of conidiation by light in Aspergillus nidulans. AB - Light regulates several aspects of the biology of many organisms, including the balance between asexual and sexual development in some fungi. To understand how light regulates fungal development at the molecular level we have used Aspergillus nidulans as a model. We have performed a genome-wide expression analysis that has allowed us to identify >400 genes upregulated and >100 genes downregulated by light in developmentally competent mycelium. Among the upregulated genes were genes required for the regulation of asexual development, one of the major biological responses to light in A. nidulans, which is a pathway controlled by the master regulatory gene brlA. The expression of brlA, like conidiation, is induced by light. A detailed analysis of brlA light regulation revealed increased expression after short exposures with a maximum after 60 min of light followed by photoadaptation with longer light exposures. In addition to brlA, genes flbA-C and fluG are also light regulated, and flbA-C are required for the correct light-dependent regulation of the upstream regulator fluG. We have found that light induction of brlA required the photoreceptor complex composed of a phytochrome FphA, and the white-collar homologs LreA and LreB, and the fluffy genes flbA-C. We propose that the activation of regulatory genes by light is the key event in the activation of asexual development by light in A. nidulans. PMID- 21624999 TI - The relation between reproductive value and genetic contribution. AB - What determines the genetic contribution that an individual makes to future generations? With biparental reproduction, each individual leaves a "pedigree" of descendants, determined by the biparental relationships in the population. The pedigree of an individual constrains the lines of descent of each of its genes. An individual's reproductive value is the expected number of copies of each of its genes that is passed on to distant generations conditional on its pedigree. For the simplest model of biparental reproduction (analogous to the Wright-Fisher model), an individual's reproductive value is determined within ~10 generations, independent of population size. Partial selfing and subdivision do not greatly slow this convergence. Our central result is that the probability that a gene will survive is proportional to the reproductive value of the individual that carries it and that, conditional on survival, after a few tens of generations, the distribution of the number of surviving copies is the same for all individuals, whatever their reproductive value. These results can be generalized to the joint distribution of surviving blocks of the ancestral genome. Selection on unlinked loci in the genetic background may greatly increase the variance in reproductive value, but the above results nevertheless still hold. The almost linear relationship between survival probability and reproductive value also holds for weakly favored alleles. Thus, the influence of the complex pedigree of descendants on an individual's genetic contribution to the population can be summarized through a single number: its reproductive value. PMID- 21625000 TI - Cell polarity in Saccharomyces cerevisiae depends on proper localization of the Bud9 landmark protein by the EKC/KEOPS complex. AB - In diploid Saccharomyces cerevisiae cells, bud-site selection is determined by two cortical landmarks, Bud8p and Bud9p, at the distal and proximal poles, respectively. Their localizations depend on the multigenerational proteins Rax1p/Rax2p. Many genes involved in bud-site selection were identified previously by genome-wide screening of deletion mutants, which identified BUD32 that causes a random budding in diploid cells. Bud32p is an atypical kinase involved in a signaling cascade of Sch9p kinase, the yeast homolog of Akt/PKB, and a component of the EKC/KEOPS (endopeptidase-like, kinase, chromatin-associated/kinase, putative endopeptidase, and other proteins of small size) complex that functions in telomere maintenance and transcriptional regulation. However, its role in bipolar budding has remained unclear. In this report, we show that the Sch9p kinase cascade does not affect bipolar budding but that the EKC/KEOPS complex regulates the localization of Bud9p. The kinase activity of Bud32p, which is essential for the functions of the EKC/KEOPS complex but is not necessary for the Sch9p signaling cascade, is required for bipolar bud-site selection. BUD9 is necessary for random budding in each deletion mutant of EKC/KEOPS components, and RAX2 is genetically upstream of EKC/KEOPS genes for the regulation of bipolar budding. The asymmetric localization of Bud9p was dependent on the complex, but Bud8p and Rax2p were not. We concluded that the EKC/KEOPS complex is specifically involved in the regulation of Bud9p localization downstream of Rax1p/Rax2p. PMID- 21625001 TI - Insight into the mechanism of nucleosome reorganization from histone mutants that suppress defects in the FACT histone chaperone. AB - FACT (FAcilitates Chromatin Transcription/Transactions) plays a central role in transcription and replication in eukaryotes by both establishing and overcoming the repressive properties of chromatin. FACT promotes these opposing goals by interconverting nucleosomes between the canonical form and a more open reorganized form. In the forward direction, reorganization destabilizes nucleosomes, while the reverse reaction promotes nucleosome assembly. Nucleosome destabilization involves disrupting contacts among histone H2A-H2B dimers, (H3 H4)(2) tetramers, and DNA. Here we show that mutations that weaken the dimer:tetramer interface in nucleosomes suppress defects caused by FACT deficiency in vivo in the yeast Saccharomyces cerevisiae. Mutating the gene that encodes the Spt16 subunit of FACT causes phenotypes associated with defects in transcription and replication, and we identify histone mutants that selectively suppress those associated with replication. Analysis of purified components suggests that the defective version of FACT is unable to maintain the reorganized nucleosome state efficiently, whereas nucleosomes with mutant histones are reorganized more easily than normal. The genetic suppression observed when the FACT defect is combined with the histone defect therefore reveals the importance of the dynamic reorganization of contacts within nucleosomes to the function of FACT in vivo, especially to FACT's apparent role in promoting progression of DNA replication complexes. We also show that an H2B mutation causes different phenotypes, depending on which of the two similar genes that encode this protein are altered, revealing unexpected functional differences between these duplicated genes and calling into question the practice of examining the effects of histone mutants by expressing them from a single plasmid-borne allele. PMID- 21625002 TI - Genetic draft and quasi-neutrality in large facultatively sexual populations. AB - Large populations may contain numerous simultaneously segregating polymorphisms subject to natural selection. Since selection acts on individuals whose fitness depends on many loci, different loci affect each other's dynamics. This leads to stochastic fluctuations of allele frequencies above and beyond genetic drift-an effect known as genetic draft. Since recombination disrupts associations between alleles, draft is strong when recombination is rare. Here, we study a facultatively outcrossing population in a regime where the frequency of outcrossing and recombination, r, is small compared to the characteristic scale of fitness differences sigma. In this regime, fit genotypes expand clonally, leading to large fluctuations in the number of recombinant offspring genotypes. The power law tail in the distribution of the latter makes it impossible to capture the dynamics of draft by an effective neutral model. Instead, we find that the fixation time of a neutral allele increases only slowly with the population size but depends sensitively on the ratio r/sigma. The efficacy of selection is reduced dramatically and alleles behave "quasi-neutrally" even for Ns?1, provided that |s| < s(c), where s(c) depends strongly on r/sigma, but only weakly on population size N. In addition, the anomalous fluctuations due to draft change the spectrum of (quasi)-neutral alleles from f(nu) ~ nu(-1), corresponding to drift, to ~ nu(-2). Finally, draft accelerates the rate of two-step adaptations through deleterious intermediates. PMID- 21625003 TI - Anterior-posterior axis specification in Drosophila oocytes: identification of novel bicoid and oskar mRNA localization factors. AB - The Drosophila melanogaster anterior-posterior axis is established during oogenesis by the localization of bicoid and oskar mRNAs to the anterior and posterior poles of the oocyte. Although genetic screens have identified some trans-acting factors required for the localization of these transcripts, other factors may have been missed because they also function at other stages of oogenesis. To circumvent this problem, we performed a screen for revertants and dominant suppressors of the bicaudal phenotype caused by expressing Miranda-GFP in the female germline. Miranda mislocalizes oskar mRNA/Staufen complexes to the oocyte anterior by coupling them to the bicoid localization pathway, resulting in the formation of an anterior abdomen in place of the head. In one class of revertants, Miranda still binds Staufen/oskar mRNA complexes, but does not localize to the anterior, identifying an anterior targeting domain at the N terminus of Miranda. This has an almost identical sequence to the N terminus of vertebrate RHAMM, which is also a large coiled-coil protein, suggesting that it may be a divergent Miranda ortholog. In addition, we recovered 30 dominant suppressors, including multiple alleles of the spectroplakin, short stop, a lethal complementation group that prevents oskar mRNA anchoring, and a female sterile complementation group that disrupts the anterior localization of bicoid mRNA in late oogenesis. One of the single allele suppressors proved to be a mutation in the actin nucleator, Cappuccino, revealing a previously unrecognized function of Cappuccino in pole plasm anchoring and the induction of actin filaments by Long Oskar protein. PMID- 21625004 TI - The Rho1 GTPase acts together with a vacuolar glutathione S-conjugate transporter to protect yeast cells from oxidative stress. AB - Maintenance of redox homeostasis is critical for the survival of all aerobic organisms. In the budding yeast Saccharomyces cerevisiae, as in other eukaryotes, reactive oxygen species (ROS) are generated during metabolism and upon exposure to environmental stresses. The abnormal production of ROS triggers defense mechanisms to avoid the deleterious consequence of ROS accumulation. Here, we show that the Rho1 GTPase is necessary to confer resistance to oxidants in budding yeast. Temperature-sensitive rho1 mutants (rho1(ts)) are hypersensitive to oxidants and exhibit high accumulation of ROS even at a semipermissive temperature. Rho1 associates with Ycf1, a vacuolar glutathione S-conjugate transporter, which is important for heavy metal detoxification in yeast. Rho1 and Ycf1 exhibit a two-hybrid interaction with each other and form a bimolecular fluorescent complex on the vacuolar membrane. A fluorescent-based complementation assay suggests that the GTP-bound Rho1 associates with Ycf1 and that their interaction is enhanced upon exposure to hydrogen peroxide. The rho1(ts) mutants also exhibit hypersensitivity to cadmium, while cells carrying a deletion of YCF1 or mutations in a component of the Pkc1-MAP kinase pathway exhibit little or minor sensitivity to oxidants. We thus propose that Rho1 protects yeast cells from oxidative stress by regulating multiple downstream targets including Ycf1. Since both Rho1 and Ycf1 belong to highly conserved families of proteins, similar mechanisms may exist in other eukaryotes. PMID- 21625006 TI - Homologous pairing and the role of pairing centers in meiosis. AB - Homologous pairing establishes the foundation for accurate reductional segregation during meiosis I in sexual organisms. This Commentary summarizes recent progress in our understanding of homologous pairing in meiosis, and will focus on the characteristics and mechanisms of specialized chromosome sites, called pairing centers (PCs), in Caenorhabditis elegans and Drosophila melanogaster. In C. elegans, each chromosome contains a single PC that stabilizes chromosome pairing and initiates synapsis of homologous chromosomes. Specific zinc-finger proteins recruited to PCs link chromosomes to nuclear envelope proteins--and through them to the microtubule cytoskeleton--thereby stimulating chromosome movements in early prophase, which are thought to be important for homolog sorting. This mechanism appears to be a variant of the 'telomere bouquet' process, in which telomeres cluster on the nuclear envelope, connect chromosomes through nuclear envelope proteins to the cytoskeleton and lead chromosome movements that promote homologous synapsis. In Drosophila males, which undergo meiosis without recombination, pairing of the largely non-homologous X and Y chromosomes occurs at specific repetitive sequences in the ribosomal DNA. Although no other clear examples of PC-based pairing mechanisms have been described, there is evidence for special roles of telomeres and centromeres in aspects of chromosome pairing, synapsis and segregation; these roles are in some cases similar to those of PCs. PMID- 21625005 TI - Assessment of the safety and biodistribution of a regulated AAV2 gene transfer vector after delivery to murine submandibular glands. AB - Clinical gene transfer holds promise for the treatment of many inherited and acquired disorders. A key consideration for all clinical gene transfer applications is the tight control of transgene expression. We have examined the safety and biodistribution of a serotype 2, recombinant adeno-associated viral (AAV2) vector that encodes a rapamycin-responsive chimeric transcription factor, which regulates the expression of a therapeutic transgene (human erythropoietin [hEpo]). The vector, AAV2-TF2.3w-hEpo (2.5 * 10(7)-2.5 * 10(10) particles), was administered once to a single submandibular gland of male and female mice and mediated hEpo expression in vivo following a rapamycin injection but not in its absence. Control (saline treated) and vector-treated animals maintained their weight, and consumed food and water, similarly. Vector delivery led to no significant toxicological effects as judged by hematology, clinical chemistry, and gross and microscopic pathology evaluations. On day 3 after vector delivery, vector copies were not only abundant in the targeted right submandibular gland but also detected in multiple other tissues. Vector was cleared from the targeted gland much more rapidly in female mice than in male mice. Overall, our results are consistent with the notion that administration of the AAV2-TF2.3w-hEpo vector to salivary glands posed no significant risk in mice. PMID- 21625007 TI - Redistribution of caveolae during mitosis. AB - Caveolae form a specialized platform within the plasma membrane that is crucial for an array of important biological functions, ranging from signaling to endocytosis. Using total internal reflection fluorescence (TIRF) and 3D fast spinning-disk confocal imaging to follow caveola dynamics for extended periods, and electron microscopy to obtain high resolution snapshots, we found that the vast majority of caveolae are dynamic with lifetimes ranging from a few seconds to several minutes. Use of these methods revealed a change in the dynamics and localization of caveolae during mitosis. During interphase, the equilibrium between the arrival and departure of caveolae from the cell surface maintains the steady-state distribution of caveolin-1 (Cav1) at the plasma membrane. During mitosis, increased dynamics coupled to an imbalance between the arrival and departure of caveolae from the cell surface induces a redistribution of Cav1 from the plasma membrane to intracellular compartments. These changes are reversed during cytokinesis. The observed redistribution of Cav1 was reproduced by treatment of interphase cells with nocodazole, suggesting that microtubule rearrangements during mitosis can mediate caveolin relocalization. This study provides new insights into the dynamics of caveolae and highlights precise regulation of caveola budding and recycling during mitosis. PMID- 21625008 TI - The histone deacetylase Rpd3 regulates the heterochromatin structure of Drosophila telomeres. AB - Telomeres are specialized structures at the end of eukaryotic chromosomes that are required to preserve genome integrity, chromosome stability and nuclear architecture. Telomere maintenance and function are established epigenetically in several eukaryotes. However, the exact chromatin enzymatic modifications regulating telomere homeostasis are poorly understood. In Drosophila melanogaster, telomere length and stability are maintained through the retrotransposition of specialized telomeric sequences and by the specific loading of protecting capping proteins, respectively. Here, we show that the loss of the essential and evolutionarily conserved histone deacetylase Rpd3, the homolog of mammalian HDAC1, causes aberrant telomeric fusions on polytene chromosome ends. Remarkably, these telomere fusion defects are associated with a marked decrease of histone H4 acetylation, as well as an accumulation of heterochromatic epigenetic marks at telomeres, including histone H3K9 trimethylation and the heterochromatic protein HP2. Our work suggests that Drosophila telomere structure is epigenetically regulated by the histone deacetylase Rpd3. PMID- 21625009 TI - A Cyp2a polymorphism predicts susceptibility to NNK-induced lung tumorigenesis in mice. AB - Lung tumors from smokers as well as lung tumors from mice exposed to tobacco carcinogens such as 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), often carry mutations in K-ras, which activates downstream-signaling pathways such as PI3K/AKT/mTOR pathway. Mice with genetic deletion of one of three isoforms of AKT were used to investigate the role of AKT in mutant K-ras-induced lung tumorigenesis in mice. Although deletion of Akt1 or Akt2 decreased NNK-induced lung tumor formation by 90%, deletion of Akt2 failed to decrease lung tumorigenesis in two other mouse models driven by mutant K-ras. Genetic mapping showed that Akt2 was tightly linked to the cytochrome P450 Cyp2a locus on chromosome 7. Consequently, targeted deletion of Akt2 created linkage to a strain specific Cyp2a5 polymorphism that decreased activation of NNK in vitro. Mice with this Cyp2a5 polymorphism had decreased NNK-induced DNA adduct formation in vivo and decreased NNK-induced lung tumorigenesis. These studies support human epidemiological studies linking CYP2A polymorphisms with lung cancer risk in humans and highlight the need to confirm phenotypes of genetically engineered mice in multiple mouse strains. PMID- 21625010 TI - Imagining being somewhere else: neural basis of changing perspective in space. AB - The capacity to imagine being somewhere else and seeing the environment from a different point of view is crucial for spatial planning in daily life and for understanding the intentions, actions, and state of mind of other people. The neural bases of spatial updating of multiple object locations were investigated using functional magnetic resonance imaging. Healthy volunteers saw an array of objects on a table in a virtual reality environment and imagined movement of their own viewpoint or rotation of the array. Their memory for the locations of the objects was then tested with a change-detection task. Behavioral results confirmed the advantage for imagined viewpoint change compared with imagined array rotation of equivalent size. Encoding of object locations was associated with a network of areas, including bilateral superior and inferior parietal cortices. The precuneus was additionally activated by the demands of both viewpoint- and array rotation. The parieto-occipital sulcus/retrosplenial cortex and hippocampus were additionally activated by the demands of viewpoint rotation, while array rotation was associated with activation of the right intraparietal sulcus. These findings support a computational model of spatial memory in which parieto-occipital sulcus/retrosplenial cortex mediates spatial updating as part of a process of translation between "egocentric" and "allocentric" reference frames. PMID- 21625011 TI - On the variability of the McGurk effect: audiovisual integration depends on prestimulus brain states. AB - The McGurk effect demonstrates the influence of visual cues on auditory perception. Mismatching information from both sensory modalities can fuse to a novel percept that matches neither the auditory nor the visual stimulus. This illusion is reported in 60-80% of trials. We were interested in the impact of ongoing brain oscillations-indexed by fluctuating local excitability and interareal synchronization-on upcoming perception of identical stimuli. The perception of the McGurk effect is preceded by high beta activity in parietal, frontal, and temporal areas. Beta activity is pronounced in the left superior temporal gyrus (lSTG), which is considered as a site of multimodal integration. This area is functionally (de)coupled to distributed frontal and temporal regions in illusion trials. The disposition to fuse multisensory information is enhanced as the lSTG is more strongly coupled to frontoparietal regions. Illusory perception is accompanied by a decrease in poststimulus theta-band activity in the cuneus, precuneus, and left superior frontal gyrus. Event-related activity in the left middle temporal gyrus is pronounced during illusory perception. Thus, the McGurk effect depends on fluctuating brain states suggesting that functional connectedness of left STS at a prestimulus stage is crucial for an audiovisual percept. PMID- 21625012 TI - Emotional voice areas: anatomic location, functional properties, and structural connections revealed by combined fMRI/DTI. AB - We determined the location, functional response profile, and structural fiber connections of auditory areas with voice- and emotion-sensitive activity using functional magnetic resonance imaging (fMRI) and diffusion tensor imaging. Bilateral regions responding to emotional voices were consistently found in the superior temporal gyrus, posterolateral to the primary auditory cortex. Event related fMRI showed stronger responses in these areas to voices-expressing anger, sadness, joy, and relief, relative to voices with neutral prosody. Their neural responses were primarily driven by prosodic arousal, irrespective of valence. Probabilistic fiber tracking revealed direct structural connections of these "emotional voice areas" (EVA) with ipsilateral medial geniculate body, which is the major input source of early auditory cortex, as well as with the ipsilateral inferior frontal gyrus (IFG) and inferior parietal lobe (IPL). In addition, vocal emotions (compared with neutral prosody) increased the functional coupling of EVA with the ipsilateral IFG but not IPL. These results provide new insights into the neural architecture of the human voice processing system and support a crucial involvement of IFG in the recognition of vocal emotions, whereas IPL may subserve distinct auditory spatial functions, consistent with distinct anatomical substrates for the processing of "how" and "where" information within the auditory pathways. PMID- 21625013 TI - Early postnatal migration and development of layer II pyramidal neurons in the rodent cingulate/retrosplenial cortex. AB - The cingulate and retrosplenial regions are major components of the dorsomedial (dm) limbic cortex and have been implicated in a range of cognitive functions such as emotion, attention, and spatial memory. While the structure and connectivity of these cortices are well characterized, little is known about their development. Notably, the timing and mode of migration that govern the appropriate positioning of late-born neurons remain unknown. Here, we analyzed migratory events during the early postnatal period from ventricular/subventricular zone (VZ/SVZ) to the cerebral cortex by transducing neuronal precursors in the VZ/SVZ of newborn rats/mice with Tomato/green fluorescent protein-encoding lentivectors. We have identified a pool of postmitotic pyramidal precursors in the dm part of the neonatal VZ/SVZ that migrate into the medial limbic cortex during the first postnatal week. Time-lapse imaging demonstrates that these cells migrate on radial glial fibers by locomotion and display morphological and behavioral changes as they travel through the white matter and enter into the cortical gray matter. In the granular retrosplenial cortex, these cells give rise to a Satb2+ pyramidal subtype and develop dendritic bundles in layer I. Our observations provide the first insight into the patterns and dynamics of cell migration into the medial limbic cortex. PMID- 21625014 TI - A to Z of nutritional supplements: dietary supplements, sports nutrition foods and ergogenic aids for health and performance--Part 21. PMID- 21625015 TI - A novel insight to the functional role of Stathmin 1 in IgE-mediated activation of RBL-2H3 cells. AB - IgE-mediated cell signaling, induced by cross-linking of high affinity receptor for IgE (FcepsilonRI) in the presence of antigen (Ag), is a well known mechanism described for mast cell activation in allergy and hypersensitivity reactions, which induces a spectrum of cellular responses such as secretion and up regulation of cell surface FcepsilonRI. Although for several years IgE binding to FcepsilonRI was considered to be a passive sensitization process, the outcomes of several recent studies have revealed a variety of different cellular responses to IgE binding compared to IgE plus Antigen binding. The present study applied a functional proteomics-based approach to investigate mast cell signaling events and provided new insights to FcepsilonRI-mediated cell signaling in RBL-2H3.1 cells, and may point to the activation of alternative signaling pathways in response to IgE or IgE plus Ag. Comparative analysis by 2-D PAGE of RBL cells activated with IgE plus Ag for three and four hours compared to non-activated cells was followed by mass spectrometric protein identification and provided evidence for the induction of Stathmin 1 (STMN1) gene expression in response to IgE plus Ag activation.Complementary SDS-PAGE analysis showed a distinct up regulation of STMN1 induction in response to challenge with IgE plus Ag compared to sensitization with IgE only. Phosphoproteomics analysis gave evidence for significant increase at phosphorylation of STMN1 on ser16 after 1min, though a slight rise at 5 min, and on ser38 after 1 and 5min sensitization with IgE and a similar result was observed for 1min IgE plus Ag-activation. IgE plus Ag activation was also found to induce the phosphorylation of ser38 to a greater extent than sensitization with IgE. In contrast, IgE alone was more effective than IgE plus Ag at inducing phosphorylation of ser16. Collectively this study provides further insights into the role of stathmin 1 in FcRI-mediated activation of cells of mast cell lineage and might shed light on the diverse response of these cells to IgE or IgE plus Ag. PMID- 21625016 TI - Expression of IL-17 and COX2 gene in peripheral blood leukocytes of vitiligo patients. AB - Vitiligo is a pigmentation disorder in which inflammatory mediators such as cytokines have a pivotal role in disease's pathogenesis. Interleukin 17 (IL-17A) is a proinflammatory cytokine which is involved in the induction of several proinflamatory mediators such as cyclooxygenase 2 (COX2). The aim of this study was to investigate the gene expression of IL-17 and COX2 in peripheral blood leukocytes of vitiligo's patients.Peripheral blood leukocytes from 15 patients with vitiligo and 15 healthy controls were separated using a gradient density centrifugation method. After total RNA isolation and cDNA synthesis, IL-17 and COX2 gene expression were quantified by real-time polymerase chain reaction (PCR). There were no significant differences in IL-17 and COX2 gene expression in lymphocytes of patients with vitiligo compared with control group (p<0.05). However there was an increased IL-17 and COX2 gene expression in neutrophils of patients compared to controls, but it did not reach statistical significance (p=0.05). We could not find any differences in IL-17 and Cox2 gene expression between clinical diseases subtypes, sex and age. There was a significant correlation between IL-17 and COX2 genes expression in the neutrophils of patients (p=0.00, r=0.80). Our results showed an increased expression in IL-17 and Cox-2 genes in neurophils of patients with vitiligo. This suggested that these two factors are involved in the inflammatory process. Further studies with a larger sample size might help to establish the role of these factors in the pathogenesis of diseases. PMID- 21625017 TI - Variant Toll-like receptor4 (Asp299Gly and Thr399Ile alleles) and Toll-like receptor2 (Arg753Gln and Arg677Trp alleles) in colorectal cancer. AB - The innate immune system recognizes the presence of bacterial products through the expression of a family of membrane receptors known as Toll-like receptors (TLRs). Polymorphisms in TLRs have been shown to be associated with increased susceptibility to diseases such as inflammatory bowel disease. The aim of this study was to determine whether there was a correlation between polymorphisms of TLR4 (Asp299Gly; Thr399Ile) and TLR2 (Arg677Trp; Arg753Gln) genes and risk of colorectal cancer. DNA from 60 colorectal carcinoma patients from 3 major races in Malaysia (22 Malays, 20 Chinese and 18 Indians) and blood from 50 apparently healthy individuals were evaluated. Control group were matched to study group by race and age. The polymorphisms were determined by Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCR-RFLP). Genotyping results showed two out of sixty tumour specimens (3.3%) harbored both variant TLR4 Asp299Gly and Thr399Ile alleles. In contrast, DNA isolated from blood cells of 50 apparently healthy individuals harbored wild type TLR4. In the case of TLR2 Arg753Gln genotyping, all of the fifty normal and 60 tumours were of the wild type genotype. TLR2 Arg677Trp genotyping showed a heterozygous pattern in all samples. However, this may not be a true polymorphism of the TLR2 gene as it is likely due to a variation of a duplicated ( pseudogene) region. There was only a low incidence (2/60; 3.3%) of TLR4 polymorphism at the Asp299Gly and Thr399Ile alleles in colorectal cancer patients. All normal and tumour samples harbored the wild type TLR2 Arg753 allele. Our study suggests that variant TLR4 (Asp299Gly and Thr399Ile alleles) as well as TLR2 (Arg753Gln allele) are not associated with risk of colorectal cancer. PMID- 21625018 TI - Co-administration of chenopodium album allergens and CpG oligodeoxynucleotides effects on peripheral blood mononuclear cells of patients with Allergic Rhinitis treated with intranasal corticosteroids and antihistamines. AB - Allergic Rhinitis (AR) is one of the most common chronic diseases in the developed countries. This study was performed to investigate the effect of CpG ODN in alteration of T-helper (Th)1/Th2 balance of patients with AR treated with intranasal corticosteroids (INCs) and antihistamines. Peripheral blood mononuclear cells (PBMCs) of 20 patients with AR were isolated before and after 45 days therapy. Cytokine production (IL-4, IL-10, IL-13, IFN-gamma) and specific Ch.a IgE in response to CpG co-administration of natural chenopodium album (CpG/Ch.a) or recombinant Ch.a (CpG/rCh.a) allergen were investigated in supernatants.of cultured PBMCs using ELISA Intracellular IL-10 was also assessed in CD4+ cells using flow cytometry. Significant increase in production of IFN gamma and IL-10 and decrease in production of IL-4 were found in supernatants of cultured PBMCs activated with CPG/ch.a and CPG/rch.a. of both CpG/Ch.a and CpG/rCh.a compared to allergens alone, before and after therapy. After therapy, IFN-gamma production with CpG/Ch.a was significantly increased in comparison with before (237 vs. 44 pg/ml, p=0.001). IFN-gamma and IL-10 production with CpG/rCh.a was significantly increased after therapy compared to before (407.6 vs. 109 pg/ml, p=0.01 for IFN-gamma; 171.7 vs. 52.6 pg/ml, p=0.008 for IL-10), whilst IL 4 was significantly decreased (2.1 vs. 5.8 pg/ml, p=0.02). Intracellular IL-10 expression was also significantly increased in response to either CpG/Ch.a or CpG/rCh.a that showed intracellular assay could be more sensitive than ELISA. Also, treatment with intranasal corticosteroids and antihistamines could enhance this CpG effect, in vitro. PMID- 21625019 TI - Comparison between sensitivity of autologous skin serum test and autologous plasma skin test in patients with Chronic Idiopathic Urticaria for detection of antibody against IgE or IgE receptor (FcepsilonRIalpha). AB - Intradermal injection of autologous serum and plasma elicit a cutaneous reactivity in almost 45-60% of patients with Chronic Idiopathic Urticaria (CIU). This reactivity is associated with the presence of auto antibodies against IgE or IgE receptors. This study was carried out to compare the cutaneous reactivity of autologous serum and plasma skin tests in a series of patients with CIU for diagnosis of auto antibodies against IgE or IgE receptor. Fifty eight patients with CIU were injected intradermally with autologous serum and plasma (anticoagulated by citrate). Histamine was used as positive control and normal saline as negative control. The study group was checked by routine laboratory tests (CBC, U/A etc), allergens with skin prick tests, and serum IgE level, and auto antibodies against thyroid as well. Duration of urticaria was another factor which was assessed.There was no significant difference between positive ASST and positive APST patients for the above mentioned tests. 77.6% of the patients were Positive for APST and 65.5% were ASST positive. Duration of urticaria was longer in patients with positive ASST and APST than ASST and APST negative patients, although the difference was not statistically significant.Autologus serum skin test (ASST) and autologous plasma skin test (APST) could be used for estimation of duration and severity of urticaria and planning for the treatment. PMID- 21625020 TI - Associations between HLA-C alleles and definite Meniere's disease. AB - Both genetic and environmental factors seem to play role in the etiology of Meniere's disease (MD). Several genes may be involved in susceptibility of MD including Human Leukocyte Antigens (HLA). The associations between MD and HLA alleles have been previously studied in other populations and certain HLA alleles were shown to be predisposing. The aim of this study was to determine the association between HLA-C allele frequencies and definite MD in patients who refer to Amir-Alam otolaryngology tertiary referral center in Tehran. Patients with definite MD (N=22) enrolled according to the diagnostic criteria of American Academy of Otolaryngology-Head and Neck Surgery (AAO-HNS). Cases with all 3 symptoms of MD (Vertigo, Tinnitus and lower frequency of sensory-neural hearing loss) were included and those with suspected MD were excluded from study. HLA-Cw allele frequencies were determined in patients non-related healthy controls (N=91) using PCR -SSP. We found that the frequency of HLACw*04 was significantly higher in patients compared to the controls [P = 0.0015, OR; 20, 95% CI (3.7 196.9)]. Our results revealed that HLA-C is a genetic predisposing factor in definite MD in patients who refer to Amir-Alam otolaryngology tertiary referral center. PMID- 21625021 TI - Pet exposure and the symptoms of asthma, allergic rhinitis and eczema in 6-7 years old children. AB - Allergic diseases are frequent in children and their prevalence and severity differ in the different regions of the world. The association between pet ownership in childhood and subsequent asthma and sensitization is very controversial.In our survey conducted with standardized method (International Study of Asthma and Allergies in Childhood), 3200 children 6-7 years old were questioned regarding asthma, allergic rhinitis and eczema. The prevalence of Attacks and shortness of breath with wheezing during last 12 months in the children who had exposure to pets in the first year of life was 34.3% 'that was less than children who had not exposure (OR=3.06, 95% confidence interval [CI] 1.14-8.21, P=0.021). Also during the past 12 months the prevalence of night dry coughs, allergic rhinitis symptoms and eczema symptoms in those who had pet exposure in the first year of their life was lower than the children did not have it. However there was no significant difference in some other symptoms of asthma in two groups.Our findings suggest that pet exposure in the first year of life can have a protective effect on asthma, allergic rhinitis and eczema. PMID- 21625022 TI - IL7R and RAG1/2 genes mutations/polymorphisms in patients with SCID. AB - SCID disorder is major failure of the immune system, usually genetic. The aim of this study was on mutations detection of RAG1, RAG2, and IL7RG genes in SCID cases. Mutation detection was performed by PCR sequencing. Our results indicated that 13 mutations were found through cases which include 4 mutations in IL7R gene (T661I, I138V, T56A, C57W), 7 mutations in RAG1 (W896X, W204R, M324V, T731I, M1006V, K820R, and R249H), and 2 mutations in RAG2 gene (R229W, DeltaT251). PMID- 21625023 TI - Giant aneurysm of thoracic and proximal abdominal aorta in a patient with common variable immunodeficiency. AB - Common variable immunodeficiency (CVID) is the most common symptomatic primary immunodeficiency disease, predisposing the patients to various tissue involvement and organ damage. Here a 16-year-old boy is presented who was referred to our center with cough, dyspnea, cyanosis, and history of recurrent pneumonia. The diagnosis of CVID was made according to reduction all serum immunoglobulin levels, normal numbers of T, B and NK lymphocyte subpopulations, poor antibodies responses. Considering abnormality in heart examination and chest X-ray, echocardiography and computed tomography angiography were performed which showed large thoraco-abdominal aortic aneurysm in this patient. Although there are some reports of cardiovascular disease associated with primary antibody deficiencies, this is the first time that such large thoraco-abdominal aortic aneurysm is reported in CVID. This may be secondary to recurrent pulmonary infections or an unknown mutation process. Cardiovascular abnormalities are an entity that should be kept in mind in patients with primary immunodeficiency diseases. PMID- 21625024 TI - Quercetin as a potential anti-allergic drug: which perspectives? AB - Flavonoids polyphenolic compounds that exert many anti-inflammatory and anti microbial effects, and exhibit an anti-allergic action. Quercetin is a flavonoids that recently has raised many issues and shown evidence about its action as a potential drug to allergy. A Chinese herbal formula, known as Food Allergy Herbal Formula (FAHF) has been related with blocking of anaphylaxis to peanuts (PNA) in mouse models. Quercetin appears to possess the same potential of FAHF as a safe anti-allergic substance but it opens only a wide perspective, at the moment, due to several complex issues that hamper the possibility to use natural medicine and phytochemicals as true drugs. PMID- 21625026 TI - Editorial: European Journal of Mass Spectrometry, the Division of Mass Spectrometry of the Italian Chemical Society and the International Year of Chemistry. PMID- 21625025 TI - Investigation of matched and mismatched duplex DNA by electrospray ionization mass spectrometry. AB - In this research, the gas-phase stabilities of matched and mismatched duplex DNA were investigated by electrospray ionization-mass spectrometry (ESI-MS). The wild type p53 duplex DNA [ds1, perfectly-matched (PM) DNA] was successfully distinguished from its three mutated DNAs [double-base mismatched DNA (DM)]. Moreover, the three DM DNAs were also well discriminated from each other using ESI-MS. Results show that the gas-phase thermodynamic stability of the DM DNAs decreased as the two mismatch spots moved closer. This implies that the dissociation of DM duplexes into two single strands prefers the mode "from middle to terminals". PMID- 21625027 TI - Review: multistage mass spectrometry in quality, safety and origin of foods. AB - Quality and safety control and the validation of origin are hot issues in the production of food and its distribution, and are of primary concern to food and agriculture organization. Modern mass spectrometry (MS) provides unique, reliable and affordable methodologies to approach with a high degree of scientificity any problem which may be posed in this field. In this review the contribution of mass spectrometry to food analysis is presented aiming at providing clues on the fundamental role of the basic principles of gas-phase ion chemistry in applied research fields. Applications in proteomics, allergonomics, glycomics, metabolomics, lipidomics, food safety and traceability have been surveyed. The high level of specificity and sensitivity of the MS approach allows the characterization of food components and contaminants present at ultra-trace levels, providing a distinctive and safe validation of the products. PMID- 21625028 TI - Structure and energetics of poly(ethylene glycol) cationized by Li(+), Na(+), K(+) and Cs(+): a first-principles study. AB - Density functional theoretical methods, including several basis sets and two functional, were used to collect information on the structure and energetic parameters of poly(ethylene glycol) (PEG), also referred to as poly(ethylene oxide) (PEO), coordinated by alkali metal ions. The oligomer chain is found to form a spiral around the alkali cation, which grows to roughly two helical turns when the oligomer size increases to about the decamer for each alkali ion. Above this size, the additional monomer units do not build the spiral further for Li(+) and Na(+); instead, they form less organized segments outside or next to the initial spiral. The distance of the first layer of co-ordinating O atoms from the alkali cation is 1.9-2.15 A for Li(+), 2.3-2.5 A for Na(+), 2.75-3.2 A for K(+) and 3.5-3.8 A for Cs(+) complexes. The number of O atoms in the innermost shell is five, six, seven and eleven for Li(+), Na(+), K(+) and Cs(+). The collision cross sections with He increase linearly with the oligomer to a very good approximation. No sign of leaning towards the 2/3 power dependence characterizing spherical particles is observed. The binding energy of the cation to the oligomer increases up to polymerization degree of about 10, where it levels off for each alkali-metal ion, indicating that this is approximately the limit of the oligomer size that can be influenced by the alkali cation. The binding energy-degree of polymerization curves are remarkably parallel for the four cations. The limiting binding energy at large polymerization degrees is about 544 kJ mol(-1), 460 kJ mol(-1), 356 kJ mol(-1) and 314 kJ mol(-1) for Li, Na, K and Cs, respectively. The geometrical features are compared with the X-ray and neutron diffraction data on crystalline and amorphous phases of conducting polymers formed by alkali-metal salts and PEG. The implications of the observations concerning collision cross sections and binding energies to ion mobility spectroscopy and mass spectrometry are discussed. PMID- 21625029 TI - Gas-phase study of molecular switches based on tautomeric proton transfer. AB - The keto-enol tautomeric equilibrium in two newly developed molecular switches was studied by using various mass spectral techniques. In these two compounds, namely 4-(phenyldiazenyl)-2-(piperidin-1- ylmethyl)naphthalen-1-ol and 2 [(1,4,7,10-tetraoxa-13-azacyclopentadecan-13-yl)methyl]-4- (phenyldiazenyl)naphthalen-1-ol, the switching on/off states are achieved by a controlled shift of the tautomeric equilibrium. In the first compound, electron impact-mass spectrometry confirms that the unprotonated dye exists as an enol tautomer, while the electrospray ionization tandem mass spectrometry (ESI-MS/MS) experiment proves the clear shift to the keto tautomeric form under switching with acid addition. In the second compound, the addition of the alkali metal ions causes transition of the tautomeric equilibrium from the pure enol to the pure keto form. The ESI-MS study demonstrated better sensitivity towards lithium ions. PMID- 21625030 TI - Gas chromatography-electron ionization-mass spectrometry analysis of O,O'- dialkyl methylphosphonites for verification analysis of the Chemical Weapons Convention. AB - Gas chromatography-mass spectrometry (GC-MS) analysis of O,O'-dialkyl methylphosphonites (DAMPs) was carried out with a view to developing a database and understanding the mechanism of fragmentation. DAMPs are included in the list of schedule 2B4 chemicals of the Chemical Weapons Convention. GC-MS analysis of DAMPs and their deuterated analogs revealed that their fragmentations were dominated by alpha-cleavages, alkenyl radical loss and hydrogen rearrangements. Based on fragment ions of deuterated analogs and density functional theory calculations, the fragmentation routes were rationalized. PMID- 21625031 TI - Matrix-assisted laser desorption/ionization-post source decay fragmentation of cystine- containing amphibian peptides with novel cysteine tags. AB - Long disulphide-containing peptides brevinins 1E and 2Ec from the skin secretion of the frog Rana ridibunda were reduced and alkylated with ten novel and three known derivatizing agents. Nine of novel reagents are maleimide derivatives. The peptides were also reduced with DTT directly onto the MALDI target without alkylation. Modified samples were subjected to MALDI-PSD study. Procedures, fragmentation patterns, fragment ion signal abundances and sequence coverage for two peptides modified with thirteen tags (or on-plate reduced) are described. The fast on-plate procedure for reduction/alkylation was applied to Rana ridibunda crude secretion, providing intensive signals of derivatized peptides. The corresponding ions may be used for the MS/MS sequencing procedure. PMID- 21625032 TI - Structural and thermal properties of LaMnO3 from neutron diffraction and first principles studies. AB - Neutron diffraction experiments have been performed on powder samples of LaMnO(3) below and above the Jahn-Teller transition temperature of 750 K. Experimental investigations are assisted by density functional theory calculations. Theoretical studies are carried out for the orbitally ordered state of LaMnO(3) which allows one to compare the behavior of the orbitally ordered and disordered structures as a function of temperature. The temperature dependences of the structural parameters characterizing the Jahn-Teller distortions are reported and discussed. A gradual departure of the experimental data from theoretical predictions is observed above 650 K. In this range of temperatures, anions surrounding the Jahn-Teller active cations perform more isotropic thermal motion. The onset of structural phase transition induces a reduction of the crystal volume by about 0.4% which follows from the structural transformations yielding more regular oxygen octahedra formed above the phase transformation. It is found that above the Jahn-Teller transition the distortions of the MnO(6) octahedra are not completely removed. The non-vanishing distortions are accompanied by the lifted degeneracy of the Mn e(g) states. Weak residual distortions can be assigned to the short-range orbital order that persists within a local scale but it seems quenched on average giving rise to a disappearance of the long-range order coherency of the Jahn-Teller effect. PMID- 21625033 TI - Strain induced ferroelectricity in GdN: first-principles calculations. AB - Using first-principles density functional calculations and the generalized gradient approximation functional including the on-site Coulomb interaction of 4f orbitals, we show that ferroelectricity can be induced by appropriate epitaxial tensile strain in GdN with a simple rock-salt structure, and that the polarization is sensitive to the strain. The calculated phonon spectra of strained GdN also confirm the existence of ferroelectric polarization. In addition, the electronic structure and magnetic properties of strained GdN as a function of strain are investigated. The present work opens up the possibility of epitaxially tensioned GdN thin films as potential multiferroics. PMID- 21625034 TI - Elastic and antiferromagnetic anomalies in Pr0.48Ca0.52MnO3 as determined by resonant ultrasonic spectroscopy. AB - The Pnma incommensurate phase transition in the perovskite Pr(0.48)Ca(0.52)MnO(3) at ~ 235 K is accompanied by shear strains of up to ~ 2.5% (from neutron diffraction) and changes in the shear modulus of up to ~ 40% (from resonant ultrasound spectroscopy, RUS), indicating strong coupling between the structural order parameter and strain. In contrast, the antiferromagnetic (AFM) ordering transition at ~ 180 K displays no detectable static strain, implying that there is either no coupling or only very weak coupling between the magnetic order parameter and strain. Conventional analysis of RUS data, based on measurements of resonance peak frequencies and peak widths, also failed to detect any anomaly in elastic or anelastic properties due to the AFM ordering. A new approach to the analysis the RUS data, based on autocorrelation and convolution of the entire spectra, however, has revealed that the AFM order does indeed affect the elastic behaviour in an unexpected manner. The new analysis shows, firstly, that dynamical fluctuations of the charge density ordering at T > T(c) = 237 K lead to an increase of the RUS amplitude and of the spectral convolution function. Secondly, fluctuations and convolution effects peak at the transition point and decrease with decreasing temperatures. Below 180 K the stripe structure is essentially static. Finally, AFM ordering leads to an increase in the damping of the elastic resonances. PMID- 21625035 TI - Universal transport signatures in two-electron molecular quantum dots: gate tunable Hund's rule, underscreened Kondo effect and quantum phase transitions. AB - We review here some universal aspects of the physics of two-electron molecular transistors in the absence of strong spin-orbit effects. Several recent quantum dot experiments have shown that an electrostatic backgate could be used to control the energy dispersion of magnetic levels. We discuss how the generally asymmetric coupling of the metallic contacts to two different molecular orbitals can indeed lead to a gate-tunable Hund's rule in the presence of singlet and triplet states in the quantum dot. For gate voltages such that the singlet constitutes the (non-magnetic) ground state, one generally observes a suppression of low voltage transport, which can yet be restored in the form of enhanced cotunneling features at finite bias. More interestingly, when the gate voltage is controlled to obtain the triplet configuration, spin S = 1 Kondo anomalies appear at zero bias, with non-Fermi liquid features related to the underscreening of a spin larger than 1/2. Finally, the small bare singlet-triplet splitting in our device allows fine-tuning with the gate between these two magnetic configurations, leading to an unscreening quantum phase transition. This transition occurs between the non-magnetic singlet phase, where a two-stage Kondo effect occurs, and the triplet phase, where the partially compensated (underscreened) moment is akin to a magnetically 'ordered' state. These observations are put theoretically into a consistent global picture by using new numerical renormalization group simulations, tailored to capture sharp finite voltage cotunneling features within the Coulomb diamonds, together with complementary out-of-equilibrium diagrammatic calculations on the two-orbital Anderson model. This work should shed further light on the complicated puzzle still raised by multi-orbital extensions of the classic Kondo problem. PMID- 21625036 TI - Toward a theory for low-frequency spin dynamics in plane copper oxide superconductors: crossover from localized spins to weak coupling charge carriers with doping. AB - We explore for all wavevectors through the Brillouin zone the dynamic spin susceptibility chi(total)(+,-)(omega, q) that takes into account the interplay of localized and itinerant charge carriers. The imaginary part, Imchi(total)(+, )(omega, q), has peaks at the antiferromagnetic wavevector Q = (pi, pi) and a diffusive-like, extremely narrow and sharp peak (symmetric ring of maxima |q| = q(0)) at very small wavevectors Q(0) is proportional to w/J ~ 10(-6) with the nuclear magnetic/quadrupole resonance frequency omega and the superexchange coupling constant J. We demonstrate the capability of Imchi(total)(+,-)(omega, q) for plane copper (63)(1/T(1)) and oxygen (17)(1/T(1)) nuclear spin-lattice relaxation rate calculations from carrier free right up to optimally doped La(2 - x)Sr(x)CuO(4) and obtain the basic features of temperature and doping behavior for (63)(1/T(1)) in agreement with experimental observations. PMID- 21625037 TI - Helical magnetism and structural anomalies in triangular lattice alpha-SrCr2O4. AB - alpha-SrCr(2)O(4) has a triangular planar lattice of d(3) Cr(3+) made from edge sharing CrO(6) octahedra; the plane shows a very small orthorhombic distortion from hexagonal symmetry. With a Weiss temperature of - 596 K and a three dimensional magnetic ordering temperature of 43 K, the magnetic system is quasi two-dimensional and frustrated. Neutron powder diffraction shows that the ordered state is an incommensurate helical magnet, with an in-plane propagation vector of k = (0, 0.3217(8), 0). Temperature dependent synchrotron powder diffraction characterization of the structure shows an increase in the inter-plane spacing on cooling below 100 K and an inflection in the cell parameters at the magnetic ordering temperature. These anomalies indicate the presence of a moderate degree of magnetostructural coupling. PMID- 21625038 TI - One-dimensional pattern of Au nanodots by ion-beam sputtering: formation and mechanism. AB - Highly ordered one-dimensional arrays of nanodots, or nanobeads, are fabricated by forming nanoripples and nanodots in sequence, entirely by ion-beam sputtering (IBS) of Au(001). This demonstrates the capability of IBS for the fabrication of sophisticated nanostructures via hierarchical self-assembly. The intricate nanobead pattern ideally serves to identify the governing mechanisms for the pattern formation: nonlinear effects, especially local redeposition and surface confined transport, are essential both for the formation and the preservation of the one-dimensional order of the nanobead pattern. PMID- 21625039 TI - Surfactant free rapid synthesis of hydroxyapatite nanorods by a microwave irradiation method for the treatment of bone infection. AB - Mesoporous nanocrystalline hydroxyapatite (nHAp) rods of size 40-75 nm long and 25 nm wide (resembling bone mineral) were synthesized under microwave irradiation without using any surfactants or modifiers. The surface area and average pore size of the nHAp were found to be 32 m(2) g(-1) and 4 nm, respectively. Rifampicin (RIF) and ciprofloxacin (CPF) loaded nHAp displayed an initial burst followed by controlled release (zero order kinetics). Combination of CPF and RIF loaded nHAp showed enhanced bacterial growth inhibition against Staphylococcus aureus (S. aureus), Staphylococcus epidermidis (S. epidermidis) and Escherichia coli (E. coli) compared to individual agent loaded nHAp and pure nHAp. In addition, decreased bacterial adhesion (90%) was observed on the surface of CPF plus RIF loaded nHAp. The biocompatibility test toward MG63 cells infected with micro-organisms showed better cell viability and alkaline phosphatase activity (ALP) for the combination of CPF and RIF loaded nHAp. The influence on cell viability of infected MG63 cells was attributed to the simultaneous and controlled release of CPF and RIF from nHAp, which prevented the emergence of subpopulations that were resistant to each other. Hence, apart from the issue of the rapid synthesis of nHAp without surfactants or modifiers, the simultaneous and controlled release of dual drugs from nHAp would be a simple, non-toxic and cost-effective method to treat bone infections. PMID- 21625040 TI - ZnO and conjugated polymer bulk heterojunction solar cells containing ZnO nanorod photoanode. AB - Hybrid solar cells based on poly(3-hexylthiophene) (P3HT) and ZnO nanoparticle bulk heterojunctions (BHJ) combined with ZnO nanorod arrays were fabricated and analyzed. The dispersion of ZnO nanoparticles in P3HT is assisted by dye molecules, which function as a surface modifier for ZnO nanoparticles to improve compatibility between ZnO nanoparticles and P3HT. Compared to the ZnO nanorod/P3HT devices, the optimized cells with the ZnO nanoparticles dispersed in P3HT can significantly increase the short-circuit current and the overall power conversion efficiency from 1.36 mA cm(-2) to 2.51 mA cm(-2) and from 0.18% to 0.45% with 625 nm long ZnO nanorod arrays, respectively. The novel scheme of using the light-absorbing dye molecules both as light absorber and as surfactant for ZnO nanoparticles presents a facile route towards forming bulk heterojunction hybrid solar cells based on semiconducting nanomaterials and conjugated polymers. PMID- 21625041 TI - Colorimetric and dynamic light scattering detection of DNA sequences by using positively charged gold nanospheres: a comparative study with gold nanorods. AB - We introduce a new genosensing approach employing CTAB (cetyltrimethylammonium bromide)-coated positively charged colloidal gold nanoparticles (GNPs) to detect target DNA sequences by using absorption spectroscopy and dynamic light scattering. The approach is compared with a previously reported method employing unmodified CTAB-coated gold nanorods (GNRs). Both approaches are based on the observation that whereas the addition of probe and target ssDNA to CTAB-coated particles results in particle aggregation, no aggregation is observed after addition of probe and nontarget DNA sequences. Our goal was to compare the feasibility and sensitivity of both methods. A 21-mer ssDNA from the human immunodeficiency virus type 1 HIV-1 U5 long terminal repeat (LTR) sequence and a 23-mer ssDNA from the Bacillus anthracis cryptic protein and protective antigen precursor (pagA) genes were used as ssDNA models. In the case of GNRs, unexpectedly, the colorimetric test failed with perfect cigar-like particles but could be performed with dumbbell and dog-bone rods. By contrast, our approach with cationic CTAB-coated GNPs is easy to implement and possesses excellent feasibility with retention of comparable sensitivity--a 0.1 nM concentration of target cDNA can be detected with the naked eye and 10 pM by dynamic light scattering (DLS) measurements. The specificity of our method is illustrated by successful DLS detection of one-three base mismatches in cDNA sequences for both DNA models. These results suggest that the cationic GNPs and DLS can be used for genosensing under optimal DNA hybridization conditions without any chemical modifications of the particle surface with ssDNA molecules and signal amplification. Finally, we discuss a more than two-three-order difference in the reported estimations of the detection sensitivity of colorimetric methods (0.1 to 10-100 pM) to show that the existing aggregation models are inconsistent with the detection limits of about 0.1-1 pM DNA and that other explanations should be developed. PMID- 21625042 TI - Transparent conducting films from NbSe3 nanowires. AB - We have developed methods to disperse and partially size separate NbSe(3) nanowires in aqueous surfactant solutions. These dispersions can easily be formed into thin films. Optical and electrical studies show these films to display sheet resistances and transmittances ranging from (460 Omega/?, 22%) to (12 kOmega/?, 79%) depending on thickness. For thicker films, we measured the transparent conducting figure of merit to be sigma(DC, B)/sigma(Op) = 0.32, similar to graphene networks. Thickness measurements gave individual values of sigma(Op) = 17,800 S m(-1) and sigma(DC, B) = 5700 S m(-1). Films thinner than ~ 70 nm displayed reduced DC conductivity due to percolative effects. PMID- 21625043 TI - TiO2 nanotube membranes on transparent conducting glass for high efficiency dye sensitized solar cells. AB - Crack-free TiO(2) nanotube (NT) membranes were obtained by short time re anodization of a sintered TiO(2) NT array on Ti foil, followed by dilute HF etching at room temperature. The resulting freestanding TiO(2) membranes were opaque with a slight yellow color having one end open and another end closed. The membranes were then fixed on transparent fluorine-tin-oxide glass using a thin layer of screen-printed TiO(2) nanoparticles (NPs) as a binding medium. It was found that low temperature treatment of the resulting NT/NP film under appropriate pressure before sintering at 450 degrees C was critical for successful fixation of the NT membrane on the NP layer. The resulting films with open-ends of NT membranes facing the NP layer (open-ends down, OED, configuration) exhibited better interfacial contact between NTs and NPs than those with closed-ends facing the NP layer (closed-ends down, CED, configuration). The cells with an OED configuration exhibit higher external quantum efficiency, greater charge transfer resistance from FTO/TiO(2) to electrolyte, and better dye loading compared to CED configurations. The solar cells with the OED configuration gave 6.1% energy conversion efficiency under AM1.5G condition when the commercial N719 was used as a dye and I(-)/I(3)(-) as a redox couple, showing the promise of this method for high efficiency solar cells. PMID- 21625044 TI - JMNI special issue--tendons: the connection between muscle and bone. PMID- 21625045 TI - Involvement of proteoglycans in tendinopathy. AB - A major feature of chronic tendinopathy is a change in the nature and organisation of the extracellular matrix of tendon. Increased levels of proteoglycans have been shown in the extracellular matrix of tendinopathic tendons and these appear to influence the increased hydration and swelling of the tissue that is a feature of this condition. There is a paucity of knowledge about proteoglycans in normal and tendinopathic tendons. This review sets out to describe the nature, function and metabolism of proteoglycans present in normal tendon and in tendinopathy and outlines how changes in proteoglycan metabolism may contribute to the development and progression of this disease. PMID- 21625046 TI - The interface of mechanical loading and biological variables as they pertain to the development of tendinosis. AB - Different tendons are designed to withstand different mechanical loads in their individual environments. Variable physiologic loading ranges and correspondingly different injury thresholds lead to tendon heterogeneity. Also, tendon heterogeneity is evident when examining how different tendons regulate their response to changes in mechanical loading (over- and under-loading). The response of tendons to changes in mechanical loading plays an important role in the induction and progression of tendinosis which is tendon degeneration without inflammation. Tendon overuse injury is likely related to abnormal mechanical loading that deviates from normal mechanical loading in magnitude, frequency, duration and/or direction. Mechanical loading that results in tendon overuse injury can initiate a repair process but, after failed initial repair, non resolving chronic attempted repair appears to lead to a "smoldering" fibrogenesis. Contributions of regulatory components, including minor components in the "nerve-mast cell-myofibroblast axis", are key features in the development and progression of tendinosis. Hormonal and genetic factors may also influence risk for tendinosis. Further understanding of how tendinosis induction is related to mechanical use/overuse, how tendinosis progression is related to abnormal regulation of attempted repair, and how induction and/or progression are modulated by other risk factors may lead to interventions that mitigate risk and enhance functional repair. PMID- 21625047 TI - Tendon fatigue in response to mechanical loading. AB - Tendinopathies are commonly attributable to accumulation of sub-rupture fatigue damage from repetitive use. Data is limited to late stage disease from patients undergoing surgery, motivating development of animal models, such as ones utilizing treadmill running or repetitive reaching, to investigate the progression of tendinopathies. We developed an in vivo model using the rat patellar tendon that allows control of the loading directly applied to the tendon. This manuscript discusses the response of tendons to fatigue loading and applications of our model. Briefly, the fatigue life of the tendon was used to define low, moderate and high levels of fatigue loading. Morphological assessment showed a progression from mild kinks to fiber disruption, for low to high level fatigue loading. Collagen expression, 1 and 3 days post loading, showed more modest changes for low and moderate than high level fatigue loading. Protein and mRNA expression of Ineterleukin-1beta and MMP-13 were upregulated for moderate but not low level fatigue loading. Moderate level (7200 cycles) and 100 cycles of fatigue loading resulted in a catabolic and anabolic molecular profile respectively, at both 1 and 7 days post loading. Results suggest unique mechanisms for different levels of fatigue loading that are distinct from laceration. PMID- 21625048 TI - In vivo investigation of tendon responses to mechanical loading. AB - Tendons transmit skeletal muscle forces to bone and are essential in all voluntary movement. In turn, movement appears to affect tendon properties, and in recent years considerable effort has been put into discovering how tendon tissue responds to mechanical stimuli in vivo. Months and years of mechanical loading can influence the gross morphology of tendon, seen as an increase tendon cross sectional area (CSA). Similarly, tendon stiffness appears to be affected by weeks to months of loading. Increased stiffness can relate to changes in CSA and/or tendon material properties (modulus), though the relative contribution of these parameters is largely unclear. The possible mechanisms behind alterations in tendon material properties include changes in collagen fibril morphology and levels of cross-linking between collagen molecules. Furthermore, increased levels of collagen synthesis and expression are seen as a response to acute exercise and training, and may be a central parameter in tendon adaptation to loading. There are indications that this collagen-induction relates to the auto-/paracrine action of collagen-stimulating growth factors, such as TGFbeta-1 and IGF-I, which are expressed in response to mechanical stimuli. PMID- 21625049 TI - Mechanical force modulates scleraxis expression in bioartificial tendons. AB - Following tendon injury, cartilage, bone and fat metaplasia are often observed, making the optimization of tenocyte differentiation an important clinical goal. In this study we examined the effect of static and cyclic mechanical loading on the expression of genes which play a role in tenocyte differentiation and function, namely scleraxis (Scx) and Type I collagen (Col1a1), and determined the effect of varying mechanical parameters including (1) static vs dynamic load, (2) increasing strain magnitude, (3) inclusion of 10 s rest periods, and (4) increasing cycle number. Cyclic loading resulted in a greater increase of tenocyte gene expression than static loading over 3 weeks in culture. Increasing strain levels potentiated the induction of tenocyte genes. The insertion of a 10 s rest periods further enhanced tenocyte gene expression, as did increasing repetition numbers. These results suggest that mechanical signaling exerts an important influence on the expression of genes which play a role in determining the tendon phenotype. Further work is required to confirm and extend these findings in primary cells such as resident tendon progenitor/stem cells, in order to provide an improved understanding of biology from which optimized rehabilitation programs can be developed. PMID- 21625050 TI - Endogenous substance P production in the Achilles tendon increases with loading in an in vivo model of tendinopathy-peptidergic elevation preceding tendinosis like tissue changes. AB - OBJECTIVES: To quantify the intratendinous levels of substance P (SP) at different stages of overload in an established model for Achilles tendinopathy (rabbit). Also, to study the distribution of the SP-receptor, the NK-1R, and the source of SP, in the tendon. METHODS: Animals were subjected to the overuse protocol for 1, 3 or 6 weeks. One additional group served as unexercised controls. Immunoassay (EIA), immunohistochemistry (IHC), and in situ hybridisation (ISH) were performed. RESULTS: EIA revealed increased SP-levels in the Achilles tendon of the exercised limb in all the experimental groups as compared to in the controls (statistically significant; p=0.01). A similar trend in the unexercised Achilles tendon was observed but was not statistically significant (p=0.14). IHC and in ISH illustrated reactions of both SP and NK-1R mainly in blood vessel walls, but the receptor was also found on tenocytes. CONCLUSIONS: Achilles tendon SP-levels are elevated already after 1 week of loading. This shows that increased SP-production precedes tendinosis, as tendinosis-like changes occur only after a minimum of 3 weeks of exercise, as shown in a recent study using this model. We propose that central neuronal mechanism may be involved as similar trends were observed in the contralateral Achilles tendon. PMID- 21625051 TI - Models for the study of tendinopathy. AB - Tendinopathy refers to the clinical presentation of activity-related pain, focal tendon tenderness, and intratendinous imaging changes. The underlying pathology was once thought to be due to inflammation ('tendinitis'), but is now considered to predominantly result from degeneration ('tendinosis'). While some progress has been made in understanding tendinosis, the condition remains poorly understood and a need exists for suitable exploratory preclinical models. It is unlikely that one suitable model exists because of the complexity of the underlying pathology and myriad of possible causes. This paper provides an overview of current models utilized in tendinopathy research. It progresses hierarchically from in vitro and ex vivo models to in vivo models. For each model, rationale for use, pertinent findings, and advantages and disadvantages are discussed. By improving on these models, new methods for the prevention and treatment of tendinopathy may be explored with the ultimate outcome being a reduction in the occurrence and effects of the condition in humans. PMID- 21625052 TI - Rotator cuff tears: what have we learned from animal models? AB - Rotator cuff tendon tears are among the most common soft tissue injuries that occur at the shoulder. Despite advancements in surgical repair techniques, rotator cuff repairs experience a high rate of failure. The common occurrence of tears and the frequency of re-tears require a further understanding of the mechanisms associated with injuries, healing, and regeneration of the rotator cuff. This paper reviews in vivo studies using the various animal shoulder models of the rat, rabbit, sheep, canine, and primate. These animal models have been used to study intrinsic and extrinsic factors leading to shoulder degeneration, various suture techniques, effects of post-surgical treatment, numerous biologic and synthetic scaffolds, and an assortment of biologic augmentations used to accelerate healing. These effects can be examined in a controlled manner using animal models without other confounding factors that sometimes limit clinical studies. The clinically impactful results will be explained to highlight the specific knowledge gained from using animal models in rotator cuff research. PMID- 21625054 TI - Injectable agents derived from or targeting vascularity: has clinical acceptance in managing tendon disorders superseded scientific evidence? AB - OBJECTIVES: To report outcomes after sclerosing, platelet-rich plasma (PRP) and autologous blood injection therapies as a treatment for tendinopathy. METHODS: We searched Pubmed for clinical trials on sclerosing, PRP and autologous blood injections for tendinopathy. We scored the quality of the studies using a modified Coleman Methodological Score (CMS) with 9 criteria, which results in a final score between 0 and 90. RESULTS: We included 14 studies involving 328 tendons on sclerosing (mean CMS: 52; range 31-77), 6 studies involving 143 tendons on PRP (CMS: 57; 43-73) and 5 studies involving 160 tendons on autologous blood injections (CMS: 58, 50-68). Across treatments, the results appear promising, but as reflected by the low methodology scores, the majority of studies are non-randomized, retrospective, with small sample size or of short duration. Two of three RTCs on sclerosing injections reported better outcomes in the treatment group, while two RCTs on PRP injections show conflicting results. The only available RTC on autologous blood injections has only 8 weeks follow-up. CONCLUSIONS: There is a need for large-scale RTCs with appropriate follow-up and study size to determine the efficacy of sclerosing, platelet-rich plasma and autologous blood injection therapies as a treatment for tendinopathy. PMID- 21625053 TI - Tendon tissue engineering: progress, challenges, and translation to the clinic. AB - The tissue engineering field has made great strides in understanding how different aspects of tissue engineered constructs (TECs) and the culture process affect final tendon repair. However, there remain significant challenges in developing strategies that will lead to a clinically effective and commercially successful product. In an effort to increase repair quality, a better understanding of normal development, and how it differs from adult tendon healing, may provide strategies to improve tissue engineering. As tendon tissue engineering continues to improve, the field needs to employ more clinically relevant models of tendon injury such as degenerative tendons. We need to translate successes to larger animal models to begin exploring the clinical implications of our treatments. By advancing the models used to validate our TECs, we can help convince our toughest customer, the surgeon, that our products will be clinically efficacious. As we address these challenges in musculoskeletal tissue engineering, the field still needs to address the commercialization of products developed in the laboratory. TEC commercialization faces numerous challenges because each injury and patient is unique. This review aims to provide tissue engineers with a summary of important issues related to engineering tendon repairs and potential strategies for producing clinically successful products. PMID- 21625055 TI - A long term study on the role of exogenous human recombinant basic fibroblast growth factor on the superficial digital flexor tendon healing in rabbits. AB - BACKGROUND: This study was designed to investigate the effects of basic fibroblast growth factor on the remodeling phase of the tenotomized superficial digital flexor tendon in rabbits. METHODS: Forty white New Zealand mature male rabbits were divided randomly into two equal groups of treated and control. After tenotomy and surgical repair, using modified Kessler technique and running pattern, the injured legs were casted for 14 days. Human recombinant basic fibroblast growth factor (bFGF) was injected subcutaneously over the lesion on days 3, 7 and 10 post injuries. The control animals received normal saline injection similarly. The weight of the animals, tendon diameter, radiographic and ultrasonographic evaluations was conducted at weekly intervals. The animals were euthanized 84 days post-injury and the tendons were evaluated at macroscopic, histopathologic and ultrastructural level and were also assessed for biomechanical and percentage dry weight parameters. RESULTS: Treatment significantly reduced the diameter and increased the echogenicity and dry weight content of the injured tendons. Treatment also significantly enhanced the maturation of the tenoblasts, fibrillogenesis, the collagen fibrils' diameter, fibrillar density, stiffness, and ultimate and yield strength. CONCLUSIONS: Subcutaneous administration of human recombinant bFGF is effective in restoring the morphological and biomechanical properties of the injured SDFT in rabbits. PMID- 21625056 TI - Children with low muscle strength are at an increased risk of fracture with exposure to exercise. AB - OBJECTIVES: To use objective measures of physical fitness and muscle function to assess the interplay between exercise, muscle and fractures during childhood. METHODS: A cross-sectional analysis was performed using The Young Hearts Project, a population-based cohort recruited from Northern Ireland. Grip strength was assessed with a hand-held dynamometer. Aerobic fitness was assessed using the 20 metre endurance shuttle run. The outcome of interest was reported fractures. Data were also collected on other potential confounders. RESULTS: There were 787 boys (49.5%) and 803 girls aged 13.9+/-1.5 years. 414 (26.0%) children reported a fracture at anytime since birth. There was a positive association between higher aerobic fitness and reported fracture (OR 1.23, 95%CI 1.05 to 1.45, P=0.012) greatest in those with lowest grip strength (OR 2.10, 95%CI 1.23 to 3.31, P=0.005). Conversely, in those with highest grip strength, no association was seen between aerobic fitness and reported fractures. CONCLUSION: In children, higher levels of aerobic fitness are associated with an increased risk of fractures, with the greatest risk seen in those with low muscle strength. Our results suggest that there is the potential for exercise protocols that aim to strengthen forearm musculature to reduce upper limb fractures in adolescents. PMID- 21625057 TI - Treatment of children scoliosis by corrective brace with regulated force effect. AB - Rigid structural spine scoliosis of a child and even non progressive congenital scoliosis (e.g. isolated hemivertebra) can be treated by hypercorrective brace in full day regime. The article shows the new type of corrective brace with adjustable force effect. The brace consists of 3 stiff parts connected by joints and telescopes. The parts of brace are made from plastic according to plaster form of child trunk. The joints allow only mutual turning brace parts at frontal plane. The special telescopes were developed which operated with prescribed forces, it means the brace and trunk parts are mutually turned at prescribed moments. The article shows the algorithm for calculation of spine stress state, and spine curve correction for given brace with adjusted telescope forces. The second algorithm calculates the telescope forces for demanded spine curve correction. The computer program can be used for computer aid design of brace forces. The force effect of the new type of brace is demonstrated on a 14 months old boy with congenital scoliosis of lumbar spine (hemivertebra L1 and L3 on the right side). Curvature measured according to Coob was changed after application of this brace from 47.5 degrees to 32.0 degrees . PMID- 21625058 TI - Atypical fractures of the femur and ulna and complications of fracture healing in a 76-year-old woman with Sjogren's syndrome. PMID- 21625059 TI - Adult case of Langerhans cell histiocytosis with single site vertebral involvement. PMID- 21625060 TI - Perinatal environment and endometriosis. AB - BACKGROUND/AIMS: Perinatal environmental exposure may affect fetal development and reprogram the developing organism for adult-onset disease. In this case control study, we aimed at assessing this pathogenetic model in endometriosis. METHODS: Consecutive patients with a first laparoscopic diagnosis of endometriosis were selected as cases. Controls were women who underwent laparoscopy during the same study period, but who were found to be free of the disease. Selected women and their mothers were interviewed. RESULTS: Ninety-one women with endometriosis and 82 controls were selected. Handedness, a variable believed to be determined prenatally by hormonal environment in utero significantly differed between the study groups. Women with the disease were less likely to be left- or mixed-handed (adjusted OR: 0.24, 95% CI: 0.08-0.71). In contrast, we failed to detect any association with birth order, maternal age, smoking, nausea, weight gain, prematurity, birth weight and breast-feeding. CONCLUSIONS: Our results generally do not support the view that in utero exposure may play a major role in the pathogenesis of endometriosis. The association with handedness, however, is intriguing in this regard and deserves further investigation. PMID- 21625061 TI - Evolutionary origins of animal skeletal biomineralization. AB - The evolutionary history of biomineralization in animals is crucial to our understanding of modern mineralized tissues. Traditional methods of unravelling this history have aimed to derive a theory of the development of biomineralization through evolution by the comparison of mineralized systems in model organisms. This has led to the recognition of the 'biomineralization toolkit' and raised the question of the homology of mineralized tissues versus convergent or parallel evolution. The 'new animal phylogeny' reveals that many of the groups known to biomineralize sit among close relatives that do not, and it favours an interpretation of convergent or parallel evolution for biomineralization in animals. In addition, the fossil record of the earliest mineralized skeletons presents a rapid proliferation of biomineralization across a range of animal phyla with fossil representatives of many modern biomineralizing phyla. A synthesis of molecular, developmental, phylogenetic and fossil evidence demonstrates the convergent or parallel evolution of biomineralization in animals at the phylum level. The fossil record of the Cambrian explosion not only provides vital evidence for the evolution of animal mineralized tissues but also suggests a mechanism for its rapid and synchronous convergent origin. PMID- 21625062 TI - Eliminating exposure to aqueous solvents is necessary for the early detection and ultrastructural elemental analysis of sites of calcium and phosphorus enrichment in mineralizing UMR106-01 osteoblastic cultures. AB - The mechanism underlying the mineralization of bone is well studied and yet it remains controversial. Inherent difficulties of imaging mineralized tissues and the aqueous solubility of calcium and phosphate, the 2 ions which combine to form bone mineral crystals, limit current analyses of labile diffusible, amorphous, and crystalline intermediates by electron microscopy. To improve the retention of calcium and phosphorus, we developed a pseudo nonaqueous processing approach and used it to characterize biomineralization foci, extracellular sites of hydroxyapatite deposition in osteoblastic cell cultures. Since mineralization of UMR106-01 osteoblasts is temporally synchronized and begins 78 h after plating, we used these cultures to evaluate the effectiveness of our method when applied to cells just prior to the formation of the first mineral crystals. Our approach combines for the first time 3 well-established methods with a fourth one, i.e. dry ultrathin sectioning. Dry ultrathin sectioning with an oscillating diamond knife was used to produce electron spectroscopic images of mineralized biomineralization foci which were high-pressure frozen and freeze substituted. For comparison, cultures were also treated with conventional processing and wet sectioning. The results show that only the use of pseudo nonaqueous processing was able to detect extracellular sites of early calcium and phosphorus enrichment at 76 h, several hours prior to detection of mineral crystals within biomineralization foci. PMID- 21625063 TI - Transcriptional regulation of MSX1 natural antisense transcript. AB - Msx homeogenes play an important role in the epithelial-mesenchymal interactions leading development. Msx1 is relevant for dental and craniofacial morphogenesis, as suggested by phenotypes of Msx1 mutations in human and Msx1 KO mice. Our group showed that Msx1 gene expression can be regulated by a bidirectional transcription generating long noncoding antisense (AS) RNA the expression which is linked to the Msx1 sense (S) RNA level. Thus, the aim of the present study was to analyze the synthesis of Msx1 (AS) RNA. In vivo Msx1 AS expression analysis showed that (i) the putative promoter sequence but not the transcribed sequence was important and necessary for its expression, (ii) 2 different areas of alveolar bone can be distinguished depending on Msx1 S and AS expression, and (iii) Msx1 presence was necessary for Msx1 AS RNA full expression. In silico analysis of the Msx1 AS putative promoter region showed the presence of 4 Msx response elements possibly involved in Msx1 regulation. Msx1 constitutes an example of a bidirectionally transcribed gene giving rise to an S/AS RNA pair included in the big and growing family of AS noncoding RNAs. Our results contribute to defining a link between Msx1 S and AS RNAs resulting in a fine tuned regulatory loop of Msx1 expression. The significance of this finding is that disturbance of the balance between Msx1 S and AS RNA status may be associated with tooth agenesis and bone loss. PMID- 21625064 TI - Comprehensive skeletal phenotyping and linkage mapping in an intercross of recombinant congenic mouse strains HcB-8 and HcB-23. AB - Bone biomechanical performance is a complex trait or, more properly, an ensemble of complex traits. Biomechanical performance incorporates flexibility under loading, yield and failure load, and energy to failure; all are important measures of bone function. To date, the vast majority of work has focused on yield and failure load and its surrogate, bone mineral density. We performed a reciprocal intercross of the mouse strains HcB-8 and HcB-23 to map and ultimately identify genes that contribute to differences in biomechanical performance. Mechanical testing was performed by 3-point bending of the femora. We measured femoral diaphysis cross-sectional anatomy from photographs of the fracture surfaces. We used beam equations to calculate material level mechanical properties. We performed a principal component (PC) analysis of normalized whole bone phenotypes (17 input traits). We measured distances separating mandibular landmarks from calibrated digital photographs and performed linkage analysis. Experiment-wide alpha = 0.05 significance thresholds were established by permutation testing. Three quantitative trait loci (QTLs) identified in these studies illustrate the advantages of the comprehensive phenotyping approach. A pleiotropic QTL on chromosome 4 affected multiple whole bone phenotypes with LOD scores as large as 17.5, encompassing size, cross-sectional ellipticity, stiffness, yield and failure load, and bone mineral density. This locus was linked to 3 of the PCs but unlinked to any of the tissue level phenotypes. From this pattern, we infer that the QTL operates by modulating the proliferative response to mechanical loading. On this basis, we successfully predicted that this locus also affects the length of a specific region of the mandible. A pleiotropic locus on chromosome 10 with LOD scores displays opposite effects on failure load and toughness with LOD scores of 4.5 and 5.5, respectively, so that the allele that increases failure load decreases toughness. A chromosome 19 QTL for PC2 with an LOD score of 4.8 was not detected with either the whole bone or tissue level phenotypes. We conclude that first, comprehensive, system-oriented phenotyping provides much information that could not be obtained by focusing on bone mineral density alone. Second, mechanical performance includes inherent trade-offs between strength and brittleness. Third, considering the aggregate phenotypic data allows prediction of novel QTLs. PMID- 21625065 TI - Polyphosphates affect biological apatite nucleation. AB - While biological apatite (bone mineral) resorption is understood, from the perspective of crystallization theory, nucleation is not. The degree of saturation (Omega) describes the chemical driving force for mineral dissolution (Omega <1) or formation (Omega > 1). Omega is the ratio of the ion activity product (IAP) of free apatite (available for reaction) component ion concentrations (predominately [Ca2+] and [PO3/4-)] for apatite) and its solubility product (K(sp)). Free ion concentrations can be less than total ion concentrations if the ions form complexes, or if the ion speciation changes. Within the acidic bone resorption pit, free [PO3/4-] is reduced due to speciation into H2PO4-. This reduces IAP(bio-Ap), and Omega(bio-Ap); at Omega(bio-Ap) <1, apatite dissolves. Apatite nucleation requires Omega(bio-Ap) >1, and bioaccumulation of molar total [Ca2+] and [PO3/4-] to form the 60-70 weight percent mineral in bone tissue. This is possible with the polymerization of PO3/4 into polyphosphate [polyP: (PO3-)(n)] which reduces free [PO3/4-] while leaving total [P] unchanged. polyP forms neutral complexes by chelation with Ca2+, which further reduces free [Ca2+] and Omega(bio-Ap), yet total [Ca2+] is unchanged. In vitro experiments demonstrate reduction in free [Ca2+], free [PO3/4-], and Omega(bio-Ap) by Ca-polyP formation, while total [Ca2+] and total [P] are constant. polyP depolymerization restores free [Ca2+] and [PO3/4-] to total [Ca] and [P], and increases Omega(bio-Ap), favouring apatite nucleation. PMID- 21625066 TI - Obstructive jaundice activates nitroxidergic neurons of the vago-vagal neural circuit that regulates the hepatobiliary system in rabbits. AB - In this study, we investigated the expression of neuronal nitric oxide synthase (nNOS) and nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d), two specific enzymes for nitric oxide (NO) synthesis, in the development of liver fibrosis induced by chronic bile duct ligation (BDL) in the rabbit. We specifically studied the liver-innervated nitroxidergic neurons that originate in the nodose ganglion (NG), nucleus of the solitary tract (NTS) and dorsal motor vagal nucleus (DMV). Our data showed that BDL resulted in overexpression of NADPH d/nNOS in the NG, NTS and DMV neurons. Using densitometric analysis, we found a significant increase in NADPH-d expression as a result of BDL in the NG, NTS and DMV (72.6, 79.4 and 57.4% increase, respectively). These findings were corroborated by serum biochemistry and hepatic histopathological examination, which were influenced by NADPH-d/nNOS-generated NO in the liver following BDL. Upregulation of NADPH-d/nNOS expression may have important implications, including (1) facilitation of extrahepatic biliary parasympathetic tone that promotes gallbladder emptying of excess stagnant bile; (2) relaxation of smooth muscles of bile canaliculi thus participating in the pathogenesis of cholestasis; (3) dilation of hepatic sinusoids to counter BDL-induced intrahepatic portal hypertension in which endothelia may be damaged, and (4) alterations in hepatic metabolism, such as glycogenesis, bile formation and secretion, and bilirubin clearance. PMID- 21625067 TI - Ethnic and gender inequities in the evaluation of referred short children. AB - AIMS: To examine ethnicity and gender differences in the evaluation of referred children with short stature and to investigate adherence of the primary care evaluation to published guidelines. METHODS: Cross-sectional study in a referral center. 371 short patients aged 2-18 years were included. Outcome measures were patient's growth characteristics, final diagnosis, and prevalence of pre-referral patient data. RESULTS: The study population was composed of 239 Bedouin children and 132 Jewish children (p < 0.0001). More males (61%) than females were evaluated (p < 0.0001). There were no significant differences between males and females in age and growth parameters at the time of referral. Bedouins, males and females, were significantly shorter than their Jewish counterparts at the time of referral: height SD -2.44 +/- 0.73 and -2.62 +/- 1.05 versus -2.13 +/- 0.55 and 2.21 +/- 0.57, respectively (p < 0.05). There were no significant ethnic or gender differences in the final diagnosis. Significant deficiencies in the primary care evaluation of referred short children were found. CONCLUSIONS: We demonstrated novel ethnic- and gender-based inequities in the evaluation of referred short children. We found that the current evaluation of short stature in our area does not comply with existing guidelines. PMID- 21625068 TI - Diagnosis of glucocorticoid-remediable aldosteronism in hypertensive children. AB - OBJECTIVE: Glucocorticoid-remediable aldosteronism (GRA) is caused by the presence of a chimeric gene originating from an unequal cross-over between the CYP11B1 and CYP11B2 genes. Aldosterone suppression by dexamethasone and high 18 hydroxycortisol (18-OHF) levels have been used to differentiate GRA from the other forms of primary aldosteronism. METHODS: A dexamethasone suppression test including serum 18-OHF determination and the measurement of urinary excretion of aldosterone, its metabolites and 18-OHF were performed in 3 children of a family with primary aldosteronism. Polymerase chain reactions were performed to identify the chimeric gene. RESULTS: The chimeric gene was identified in 2 children, their mother and grandmother. The affected children had an aldosterone-to-plasma renin activity ratio >30, elevated serum 18-OHF concentration and increased urinary excretion of aldosterone, its metabolites, and 18-OHF. Post-dexamethasone concentrations of serum aldosterone and 18-OHF concentrations were suppressed. CONCLUSION: Although very rare, the possible diagnosis of GRA should be considered in all children or young adults with low-renin hypertension. Since genetic testing is more specific than biochemical testing, a definitive diagnosis can only be obtained by identification of the CYP11B1/CYP11B2 chimeric gene. PMID- 21625069 TI - Percentiles of inspiratory capacity in healthy nonsmokers: a pilot study. AB - BACKGROUND: Over the past decade inspiratory capacity (IC) has received increasing attention, especially in studies on chronic obstructive pulmonary disease. Though a few recent studies provide reference values for IC, still little is known on the distribution of its values in healthy subjects. OBJECTIVE: To estimate percentiles of IC in a sample of healthy nonsmokers by applying quantile regression. METHODS: We applied quantile regression to estimate seven percentiles of IC from 5 to 95% for a given age, height and body mass index, separately in males and females, in a sample of 411 healthy nonsmokers, aged 8-73 years, drawn from the general population in Northern Italy. RESULTS: The magnitude and statistical significance of the effect of age, height and body mass on IC varied across percentiles and between genders. When the 50th percentile of IC was compared with the predicted values of several studies, including those published by the European Community for Coal and Steel, the differences were as large as 0.86 liters in males and 0.90 liters in females. The possible advantage of the 5th percentile predicted from quantile regression over the 'traditional 5th percentile' as lower limit of normal was also illustrated. CONCLUSIONS: Inference on percentiles, rather than just the mean, of IC and other lung function measures may help in the future to better understand the effect of various risk factors, model growth curves and derive more accurate reference values. PMID- 21625070 TI - Decreased expression of liver X receptor-alpha in macrophages infected with Chlamydia pneumoniae in human atherosclerotic arteries in situ. AB - In in vitro experiments, Chlamydia pneumoniae has been shown to infect macrophages and to accelerate foam cell formation. It has been hypothesized that the C. pneumoniae infection affects foam cell formation by suppressing the expression of liver X receptors (LXR), but whether such an event occurs in human atherosclerosis is not known. In this study we examined carotid artery segments, obtained by endarterectomy, in which the presence of C. pneumoniae was confirmed by both polymerase chain reaction and immunohistochemistry. The expression of LXR alpha in macrophages infected with C. pneumoniae and macrophages that were not infected was compared using a quantitative immunohistochemical analysis. The analysis revealed a 2.2-fold reduction in the expression of LXR-alpha in C. pneumoniae-infected cells around the lipid cores in atherosclerotic plaques. In the cytoplasm of laser-capture microdissected cells that were immunopositive for C. pneumoniae, electron microscopy demonstrated the presence of structures with the appearance of elementary, reticulate and aberrant bodies of C. pneumoniae. We conclude that LXR-alpha expression is reduced in C. pneumoniae-infected macrophages in human atherosclerotic lesions which supports the hypothesis that C. pneumoniae infection might suppress LXR expression in macrophages transforming into foam cells. PMID- 21625071 TI - Pre- and postnatal imaging of a congenital hepatoblastoma. AB - Prenatal ultrasound, magnetic resonance imaging and matching postnatal computer tomography imaging findings are presented in a neonate with histologically proven congenital hepatoblastoma. Familiarity with the prenatal imaging findings of a hepatoblastoma are essential to make a reliable, specific diagnosis facilitating decision-making about the pre-, peri- and postnatal management. PMID- 21625072 TI - Mediastinal bronchial artery aneurysm mimicking a subcarinal tumor. PMID- 21625073 TI - Lung cancer in chronic hypersensitivity pneumonitis. AB - BACKGROUND: So far, the association of lung cancer with chronic hypersensitivity pneumonitis (CHP) has not been studied. OBJECTIVE: We examined the prevalence and revealed clinical features of lung cancer in CHP. METHODS: We retrospectively reviewed the medical records from 1994 through 2005 and identified 11 patients (15 lesions) with lung cancer among 104 patients with CHP. Their clinical features and histopathological findings were analyzed. RESULTS: Ten men and 1 woman with a median age of 68.9 years were included. All patients had a smoking history. The most prevalent histopathological type of lung cancer was squamous cell carcinoma (53%), and all tumors were located in the peripheral region of the lung. Four patients suffered from lung cancer after the diagnosis of CHP and 1 patient had lung cancer before the diagnosis of CHP. The histological pattern of CHP showed a predominantly usual interstitial pneumonia-like lesion. Tumors were located adjacent to honeycombing in 7 (47%) of 15 lesions, bullae in 4 (27%) lesions, and relatively normal lung in 4 lesions. CONCLUSIONS: Since the prevalence of lung cancer in CHP seems to be high (10.6%) as seen in idiopathic pulmonary fibrosis, physicians should be aware of the possible complication of lung cancer in CHP. PMID- 21625074 TI - Pseudo-Meigs' syndrome caused by a uterine leiomyosarcoma: a new clinical condition. AB - INTRODUCTION: Pseudo-Meigs' syndrome is a rare condition characterized by female genital cancer, ascites, hydrothorax and tumors other than benign ovarian tumors. CASE REPORT: A 30-year-old woman arrived at our clinic with pelvic pain and dyspnea. She underwent an exploratory laparotomy for a large pelvic mass complicated by ascites and hydrothorax. Cytological examination of the effusions was negative for malignant cells. Histological analysis of the tumor mass revealed a uterine epithelioid leiomyosarcoma. After surgery, we observed resolution of the effusions. CONCLUSION: Uterine leiomyosarcoma and pseudo-Meigs' syndrome are two rare entities. To our knowledge, there are no similar reports in the literature, and therefore we present this new clinical condition due to its high scientific evidence. PMID- 21625075 TI - Facial blaschkitis: case and review. AB - We report on a 15-year-old female with a 3-month history of a pruritic, erythematous cutaneous eruption on the right cheek and perioral area. The lesion had a linear distribution following the lines of Blaschko. Histopathological findings and direct immunofluorescence were compatible with chronic cutaneous lupus erythematosus (LE). Treatment with topical steroids and systemic antimalarial agents over 2 months showed hardly any improvement contrary to similar cases reported in the literature in the past. Histological findings move this case close to LE. However, the unusual clinical presentation as well as the resistance to antimalarial drugs do not fully allow to confirm this suspicion. Therefore, we recommend to call this new entity LE-like facial Blaschkitis of the adult. PMID- 21625076 TI - Gynecomastia as a first clinical sign of nonseminomatous germ cell tumor. AB - INTRODUCTION: Gynecomastia is a common clinical sign in several diseases. In this report we present a case of gynecomastia with underlying testicular tumor which remained misdiagnosed for a prolonged period of time. CASE REPORT: A 16-year-old adolescent noticed unilateral painless swelling of the left breast. He was referred to the Department of General Surgery and examined by a breast surgeon. A diagnosis of mastitis was made and a treatment with an oral antibiotic drug began. After failure of the initial antibiotic treatment, the patient was referred to the Department of Endocrinology and left testicular cancer was diagnosed. Unilateral high inguinal orchidectomy and subsequent chemotherapeutic treatment were performed. CONCLUSION: Primary care physicians should be aware of the possibility of a concomitant presence of gynecomastia and testicular cancer. We suggest a physical examination as well as a laboratory investigation, and testicular ultrasonography of the testes in all patients with gynecomastia. PMID- 21625077 TI - Intralesional infliximab in noninfectious cutaneous granulomas: three cases of necrobiosis lipoidica. AB - BACKGROUND: Necrobiosis lipoidica is a rare granulomatous noninfectious skin disease. Treatment of this chronic debilitating disease is of importance because ulceration of the plaques may induce important psychological and physical morbidity. OBJECTIVE: Infliximab, an anti-TNF-alpha chimeric monoclonal antibody used intravenously and intralesionally for other extradermatological granulomatous diseases including Crohn's disease and sarcoidosis, was administered by intradermal injection in necrobiosis lipoidica. The aim of this study was to evaluate the efficacy and safety profile of a locally delivered drug compared to its systemic use. PATIENTS AND METHODS: Weekly injections of intralesional infliximab for 3 weeks were followed by a 1-week treatment interruption. This treatment schedule was repeated thrice. RESULTS: Two patients who benefitted from complete treatment experienced almost complete remission for up to 18 months. The third patient, who had treatment interruptions, showed partial improvement. No serious side effects were noticed, although the injections caused pain. CONCLUSIONS: This is the first report about the efficacy and safety of a therapy consisting of intralesional injections of infliximab for a granulomatous skin disease. Although this approach was clearly effective for necrobiosis lipoidica, the disease recurred several months after treatment interruption, raising the question of the need for maintenance therapy. Further controlled long-term trials are thus necessary. PMID- 21625078 TI - Breakdown of the balanced lethals in Rhoeo: the structure of the Alethal Renner complex of the homozygotic stock of Rhoeo. AB - Fluorescence in situ hybridization, base-specific fluorescence, C-banding and silver-staining were performed to reveal the cyto-molecular constitution of the karyotype in the bivalent-forming Rhoeo spathacea concolor. It was shown that the genome of this form is almost identical to the beta-complex of the ring-forming rhoeos. In spite of some modifications in the arrangement of a few distal rDNA sites and in the amount of pericentromeric AT-rich heterochromatin, the alethal genome of the bivalent-forming Rhoeo seems segmentally unaltered. Thus, the breakdown of balanced lethals in Rhoeo is most likely uncoupled from creating new chromosome arm combinations. PMID- 21625079 TI - Association between fibroblast growth factor receptor-4 gene polymorphism and risk of prostate cancer: a meta-analysis. AB - OBJECTIVES: To examine the association between fibroblast growth factor receptor 4 (FGFR-4) genetic polymorphisms (Gly-388Arg) and prostate cancer susceptibility. METHODS: A comprehensive search was conducted to identify all case-control studies of FGFR-4 polymorphisms and prostate cancer risk. Statistical analysis was performed with the software program Stata, version 8.0, and Review Manage, version 4.2. RESULTS: A total of 4 eligible studies, including 2,618 cases and 2,305 controls, relating the FGFR-4 polymorphism to the risk of prostate cancer were identified. The overall results indicated a significant association between FGFR-4 polymorphisms and prostate cancer. In the subgroup analysis by ethnicity, a significant association was found between European descent for recessive model (random effects OR 0.82, 95% CI 0.72-0.93) and co-dominant genetic model (random effects OR 0.83, 95% CI 0.68-1.00). Sensitivity analysis also found a significant association in the recessive (random effects OR 0.68, 95% CI 0.49-0.95) or the co dominant (random effects OR 0.68, 95% CI 0.49-0.96) models. Egger's test showed that publication bias was not present in any of the comparisons. CONCLUSIONS: Gly 388Arg polymorphism of FGFR-4 most likely contributes to susceptibility to prostate cancer, especially in men of European descent. PMID- 21625080 TI - 2009 influenza A infection and acute kidney injury: incidence, risk factors, and complications. AB - BACKGROUND: 2009 influenza A has spread globally. Respiratory complications and renal failure have been the leading causes for hospitalization and critical illness. We describe the risk factors and complications of acute kidney injury (AKI) in patients with influenza A. METHODS: Observational study of adult patients tested for influenza A. Outcome measures include AKI [AKI Network (AKIN) criteria] and mortality. RESULTS: From August through December 2009, 17% (89/515) of hospitalized subjects were tested positive for influenza A. The incidence of AKI (AKIN(I-III)) was 42% (37/89) in subjects with influenza A; the majority (65%, 24/37) of whom were critically ill. Risk factors for AKI included obesity, chronic kidney disease (CKD), and elevated creatine kinase. Positive influenza A status was associated with lower AKI (AKIN(I-III)) risk compared to seronegative subjects (OR 0.5, CI 0.3-0.9). Mortality in patients with influenza A and AKI requiring dialysis was 50%. CONCLUSIONS: Obesity, CKD, and elevated creatine kinase are associated with AKI in patients with influenza A. Influenza A is not independently associated with higher incidence of AKI in hospitalized patients. AKI is an independent risk factor for mortality in patients with influenza A. PMID- 21625081 TI - Combined administration of kanamycin and furosemide does not result in loss of vestibular function in Guinea pigs. AB - Aminoglycoside antibiotics are known to damage the vestibular and auditory sensory epithelia. Although loop diuretics enhance the cochleotoxic effect of aminoglycosides, it is not known whether concomitant administration of an aminoglycoside and a loop diuretic affects the vestibular system. The aim of our study was to investigate the effect of co-administration of kanamycin and furosemide upon the otolith organs and to compare it to the known vestibulotoxic effect of gentamicin. Five guinea pigs were injected with a single dose of both kanamycin (400 mg/kg, s.c.) and furosemide (100 mg/kg, i.v.), 5 animals received gentamicin (100 mg/kg, i.p.) for 10 days, and 5 untreated animals served as controls. After 7 days, vestibular function was assessed by measuring vestibular short-latency evoked potentials (VsEPs) to linear acceleration stimuli and cochlear function by auditory brainstem responses (ABRs) to clicks. Hair cell densities were determined in phalloidin-stained whole mounts of the utricles and saccules, and in midmodiolar sections of resin-embedded cochleae. Co administration of kanamycin and furosemide had no significant effect on VsEPs and hair cell densities in the utricles and saccules were not reduced. ABR thresholds were increased to a great extent (by ~60 dB), and histologically a severe loss of cochlear hair cells was observed. The effect of gentamicin, both on vestibular and cochlear function, was just the opposite. VsEP thresholds to horizontal stimulation were elevated and suprathreshold amplitudes showed a decrease, whereas cochlear function was not reduced. With this protocol, we have a tool to selectively induce cochlear or vestibular damage, which may be of interest to researchers and clinicians alike. PMID- 21625082 TI - Secondary hyperparathyroidism in chronic dialysis patients: results of the Italian FARO survey on treatment and mortality. AB - INTRODUCTION: Vitamin D receptor activator (VDRA) therapy has been shown to be associated with reduced mortality rates in chronic kidney disease (CKD) patients with secondary hyperparathyroidism (SHPT). However, differences between VDRAs in their ability to reduce both all-cause and cardiovascular-related mortality rates are not yet fully elucidated. METHODS: The objective of the current analysis was to determine the effect of VDRA therapy on mortality in an Italian dialysis population, observed prospectively every 6 months for 18 months. Patients were investigated for all-cause and cardiovascular-related mortality risk adjusted for various demographic, clinical, and/or SHPT treatment variables. RESULTS: The cumulative probabilities of all-cause and cardiovascular-related mortality were lower for patients who received any VDRA treatment compared with those who did not (p < 0.001) regardless of all measured variables. Additionally, patients who received paricalcitol and/or cinacalcet (with or without VDRAs) compared with calcitriol showed a significant improvement in both all-cause and cardiovascular related mortality (p < 0.001). Cinacalcet with or without VDRAs was not associated with a further decrease of mortality hazard ratios compared with paricalcitol monotherapy. CONCLUSIONS: VDRA therapy (associated or not with cinacalcet) was associated with improved survival in dialysis patients, independent of demographic and clinical variables. PMID- 21625083 TI - Outcomes of patients with planned initiation of hemodialysis in the IDEAL trial. AB - BACKGROUND: In clinical practice there is considerable variation in the timing of initiation of dialysis. The IDEAL trial (Initiating Dialysis Early and Late study) showed that planned early initiation of dialysis in patients with stage 5 chronic kidney disease (CKD) was not associated with an improvement in clinical outcome, but was associated with increased costs. The predominant dialysis modality worldwide is hemodialysis (HD). This subanalysis of the IDEAL trial examined whether the timing of the initiation of dialysis in those who had chosen HD influenced survival and the occurrence of complications. METHODS: Patients on the IDEAL trial were older than 18 years and had progressive advanced CKD. They were randomly assigned to commence dialysis at an estimated glomerular filtration rate (eGFR) of 10-14 ml/min (early start) or when the eGFR was 5-7 ml/min (late start). The primary outcome was death from any cause. RESULTS: Between 2000 and 2008, 362 of the 828 patients (43.7%) randomized in the trial planned to commence HD. 322 (88.9%) of these subsequently commenced HD and 17 (4.7%) commenced peritoneal dialysis, with a median time to the initiation of dialysis of 1.63 months in the early-start group and 6.93 months in the late- start group. During a median follow-up time of 3.81 years, 50 of 171 patients in the early-start group (29.2%) and 59 in the late-start group (30.1%) died (hazard ratio with early initiation=0.97: 95% CI: 0.66-1.41; p=0.86). There was no significant difference in the frequency of cardiovascular events, infections, or access related events, but there was a significantly higher frequency of fluid and electrolyte events in the late-start group (p=0.02). CONCLUSION: In this subanalysis of the IDEAL trial, patients commencing dialysis early with stage 5 CKD for whom the planned dialysis modality was HD did not have an improvement in survival or any reduction in most clinical outcomes apart from fluid and electrolyte events. PMID- 21625084 TI - More frequent hemodialysis: what do we know? Where do we stand? AB - The high mortality of hemodialysis patients has served as a strong incentive to increasing knowledge of the possible approaches to optimizing treatment strategies in this patient population. The majority of patients worldwide usually receive hemodialysis thrice-weekly, according to calculations of dialysis adequacy. An abundance of literature and studies, starting as early as the late 1960s, has shown that an increase in dialysis frequency has beneficial impacts on outcomes such as left ventricular hypertrophy, anemia, calcium and phosphorus metabolism, and health- related quality of life. Reductions in left ventricular mass and improvements in quality of life (as measured by RAND-36) by frequent hemodialysis were recently confirmed by a large-scale randomized trial. This is the first trial on this topic which was sufficiently powered and compared the findings in patients receiving frequent hemodialysis to an adequate control group. The significant findings of this trial thus confirmed the evidence in the literature which had been accumulating over decades. This manuscript reviews previous studies on more frequent hemodialysis, including published results of the Frequent Hemodialysis Network Daily Trial. PMID- 21625085 TI - Nocturnal home hemodialysis: which of your patients should choose this modality? AB - Nocturnal home hemodialysis (HD) was conceived in the 1960s, but fell out of favor in the 1970s, only to be resurrected in a modified form in the 1990s. This modality is now used by patients on 4 continents, but still accounts for only a very small number of patients who receive chronic HD therapy. Nocturnal home HD provides a weekly Kt/V urea in the range of 4.5-6, and provides superior clearance of a number of middle molecules as well as some protein-bound and charged molecules. The first randomized trials of nocturnal home HD compared to conventional 3 times per week HD were performed in the first decade of the 21st century. In the Alberta trial, 52 patients were randomized to either nocturnal home HD conducted 5-6 times per week or conventional HD conducted 3 times per week; the patients were followed for 6 months. In this trial, subjects who received nocturnal home HD had a decrease in left ventricular mass, and improvement in the management of hypertension and hyperphosphatemia, but no significant benefit in either quality of life or anemia management. There was a decrease in systolic blood pressure, despite a decrease in the number of antihypertensive medications prescribed. There was also a decrease in serum phosphorus levels as well as a marked reduction or elimination of phosphate binders in more than two thirds of the nocturnal subjects. In the Frequent Hemodialysis Network study of Nocturnal Hemodialysis, 87 patients were randomized to either nocturnal home HD (6 times per week) or conventional (3 times per week) dialysis, and were followed for 12 months. Results from this trial will be published in 2011. It is likely that the results from these 2 trials will influence the update to the National Kidney Foundation-Kidney Disease Outcomes Quality Initiative HD guidelines that will be published in 2012. PMID- 21625086 TI - Perspectives in home hemodialysis therapy. AB - Currently, there is renewed interest in home hemodialysis. However, home hemodialysis is not for every patient--the majority will continue to need treatment away from their home in hospital or satellite units. Those who are capable of undertaking treatment at home should be strongly encouraged to do so. Treatment at home permits improved quality of life and rehabilitation, and is cost-effective. However, it needs nephrologists versed in the techniques and benefits of home hemodialysis as well as nursing staff capable of undertaking the training, underpinned by a well-functioning predialysis program geared towards identifying and recruiting patients suitable for home hemodialysis well before dialysis is required. This review explores the origins, decline and reawakened interest in this treatment modality, and discusses novel technological approaches to ease the patient's treatment burden. PMID- 21625087 TI - Lessons from recent trials on hemodialysis. AB - Today, hemodialysis (HD) represents a rescue therapy for an increasing number of patients worldwide. Thanks to continuous improvements, it is now better tolerated; thus, allowing patients relief from uremic symptoms and increasing survival. However, many questions regarding the best way of ameliorating the outcomes of chronic kidney disease patients requiring dialysis are still open. Recently, 2 randomized controlled clinical trials tried to give some answers to the current debates around dialysis. The first one--the IDEAL trial--evaluated the effects of beginning early or late dialysis on patient mortality and morbidity, and it did not find any significant difference between the 2 groups, suggesting that starting dialysis on the basis of an estimate of GFR alone is not suitable. The second one--the FHN daily trial--compared in-center conventional (3 times per week) with in-center frequent (6 times per week) HD. It found that daily dialysis is associated with improvements in left ventricular mass, physical health composite scores and some secondary outcomes (hypertension and hyperphosphatemia) - although it also discovered there had been more frequent interventions related to vascular access. Despite the fact that both studies presented some unavoidable limitations, they gave important information which is useful in everyday clinical practice. According to evidence-based medicine, such well-designed and well-conducted randomized controlled trials are the best way to improve our knowledge. PMID- 21625088 TI - The quest to standardize hemodialysis care. AB - A large global dialysis provider's core activities include providing dialysis care with excellent quality, ensuring a low variability across the clinic network and ensuring strong focus on patient safety. In this article, we summarize the pertinent components of the quality assurance and safety program of the Diaverum Renal Services Group. Concerning medical performance, the key components of a successful quality program are setting treatment targets; implementing evidence based guidelines and clinical protocols; consistently, regularly, prospectively and accurately collecting data from all clinics in the network; processing collected data to provide feedback to clinics in a timely manner, incorporating information on interclinic and intercountry variations; and revising targets, guidelines and clinical protocols based on sound scientific data. The key activities for ensuring patient safety include a standardized approach to education, i.e. a uniform education program including control of theoretical knowledge and clinical competencies; implementation of clinical policies and procedures in the organization in order to reduce variability and potential defects in clinic practice; and auditing of clinical practice on a regular basis. By applying a standardized and systematic continuous quality improvement approach throughout the entire organization, it has been possible for Diaverum to progressively improve medical performance and ensure patient safety. PMID- 21625089 TI - The burden of cardiovascular risk in chronic kidney disease and dialysis patients (cardiorenal syndrome type 4). AB - Type 4 cardiorenal syndrome is a growing societal problem as the aging population, with increasing incidence of chronic kidney disease (CKD), diabetes, obesity, hypertension, and other cardiovascular risks, leads to higher numbers of individuals suffering the complications of this bidirectional disorder. In this review, the immense burden of cardiovascular risk faced by patients on dialysis, as well as lesser degrees of CKD, will be examined, with emphasis on rates of acute coronary syndrome/coronary insufficiency, congestive heart failure/left ventricular hypertrophy, and cardiac arrhythmia/sudden cardiac death. PMID- 21625090 TI - CD14+CD16+ monocytes from chronic kidney disease patients exhibit increased adhesion ability to endothelial cells. AB - Chronic kidney disease (CKD) patients present an inflammatory process that induces endothelial damage and therefore plays a role in the high rates of cardiovascular morbidity and mortality reported in these patients. Although new therapies have reduced the elevated serum levels of inflammatory mediators such as cytokines and CRP in CKD patients, the rise in the level of activated immunocompetent cells is maintained in peripheral blood, which appears to play a prominent role in the endothelial damage suffered by these patients. CD14+CD16+ monocytes are a subset of activated monocytes that are found in greater numbers in the peripheral blood of CKD patients. The increased presence of these cells is related to the endothelial damage suffered by these patients. However, the mechanism through which these cells damage the vascular endothelium is still unclear. One of the characteristics that differentiate CD14+CD16+ monocytes is their powerful ability to produce inflammatory cytokines, which may be responsible for causing damage to endothelial cells. However, it is difficult to imagine that the cytokines produced by a relatively small proportion of these cells are capable of damaging the endothelium. For this reason, we have suggested that these cells do not release their cytokines into the bloodstream, but that they possess cellular mechanisms that lead them to produce and release cytokines after adhering to the layer of endothelial cells. This hypothesis is based on the fact that unlike the CD14++CD16- monocytes found in healthy subjects, CD14+CD16+ monocytes in CKD patients show a high level of expression of chemokines that favors their migration to the vascular wall, and a low level of chemokines such as CCR2 that would prevent such migration. Furthermore, these CD14+CD16+ monocytes express a large number of adhesion molecules, which helps them attach to endothelial cells. In view of this scenario, it is easy to suggest that a moderate number of CD14+CD16+ monocytes might well be capable of producing endothelial damage; therefore, the rise in the number of these cells in CKD patients may play an important role in the development of vascular disease. PMID- 21625091 TI - Uric acid as a cardiometabolic risk factor: to be or not to be. AB - Most authorities do not consider hyperuricemia to be an important risk factor for cardiovascular or renal disease. However, emerging data suggest that elevated uric acid (UA) is actually one of the most important factors for cardiovascular disease and that it also plays a significant role in the development of renal disease and metabolic syndrome. In the current review, we will analyze the role of cardiometabolic disorders related to UA and briefly explain the pathogenetic mechanisms behind these relationships. PMID- 21625092 TI - Treatment disparities in acute coronary syndromes, heart failure, and kidney disease. AB - It has been consistently observed that patients with renal dysfunction have more premature, severe, complicated, and fatal cardiovascular disease than age- and sex-matched individuals with normal renal function. There have been 4 major explanations for this finding: (1) positive confounding by third variables associated with chronic kidney disease (CKD), including diabetes mellitus and hypertension; (2) therapeutic nihilism or lesser use of beneficial therapies in CKD; (3) greater toxicities of therapies, such as bleeding from anticoagulants or contrast-induced kidney injury; (4) biological factors which result directly from CKD that work to promote and accelerate cardiovascular disease. In this paper, we focus on the issue of treatment disparities or therapeutic nihilism and its contribution to poor outcomes in the setting of acute coronary syndromes and acutely decompensated heart failure. This issue is important because if we can overcome barriers to the utilization of beneficial treatments, then clinical outcomes should improve over time. PMID- 21625093 TI - Hypervolemic and optivolemic natriuretic peptides in acute heart failure. AB - Acute heart failure has emerged as the leading diagnosis among hospitalized patients, and challenges in accurate diagnosis, risk stratification and optimized management still remain. Here, biomarkers--with their low cost, objectivity and widespread availability--can play an indispensible role. Among the biomarkers available, natriuretic peptides (NPs) are the most validated and accepted for risk stratification and treatment guidance. The physiological basis for this lies in the strong correlation between NP levels and pulmonary capillary wedge pressure. The ability to classify individuals on the basis of risk could allow clinicians to tailor therapies to fit individual patient needs, thus reducing morbidity, mortality and costs. PMID- 21625094 TI - Do catheters harm the patient? AB - There is an increasing use of central venous catheters (CVCs) as a first-line vascular access in incident hemodialysis (HD) patients. CVCs are associated with short-term complications related to the catheter insertion procedure and with long-term complications as a consequence of the intravascular interaction of catheter material with the venous endothelium. Removal of CVCs is also associated with specific risks, such as pulmonary or air embolism. Bacteremia and sepsis, however, are the most important risks associated with CVCs. In incident HD patients, the mortality risk from infection is highest during the first 6 months after dialysis initiation, and has steadily increased over the last decade. The relative risk for infection from CVCs in HD patients now clearly exceeds the risk for catheter-related infections in peritoneal dialysis (PD) patients. CVCs should therefore always be the last preference for vascular access in HD. The option for PD should always be evaluated before using CVCs as a permanent vascular access in end-stage renal disease patients. PMID- 21625095 TI - The evils of intradialytic sodium loading. AB - Increased salt intake is related to extracellular fluid expansion and a rise in blood pressure, and has been linked to cardiovascular disease. Several studies have also suggested that sodium can exert detrimental effects via blood pressure independent mechanisms. Chronic kidney disease patients are particularly susceptible to the negative consequences of sodium loading. While individuals with normal kidney function are able to regulate their sodium balance, hemodialysis patients have to rely virtually entirely on the dialysis procedure for sodium elimination. Tragically, the dialysis procedure has, in many instances, turned into a de facto source of sodium loading rather than a means for sodium removal. The main sources of sodium related to the dialysis procedure are (1) diffusive influx from the dialysate, including inappropriate use of sodium profiling; (2) the use of saline solution for priming and rinsing; and (3) the treatment of intradialytic hypotension and cramps with saline solution. Creating a positive intradialytic sodium balance is effective in acutely reducing the incidence of intradialytic symptoms, but it also sustains a vicious cycle hampering the attainment of dry weight and predisposes the patient to an increased risk of intradialytic complications during the following dialysis session. Avoiding sodium loading in hemodialysis patients is a cornerstone of blood pressure and fluid status management and, therefore, deserves a conscious effort, bearing in mind not only short-term effects but also long-term goals. In the absence of routine means of quantifying sodium balance, sodium profiling should be viewed critically, as it has been shown to induce a positive sodium balance in the majority of cases. A preferred approach under these circumstances may be simple sodium alignment. In combination with the abdication of saline solution for priming, rinsing, and intradialytic infusions, these measures would go a long way to help reduce sodium overload and achieve a more physiologic sodium balance in this patient population. PMID- 21625096 TI - Recent trials on hemodiafiltration. AB - The theoretical advantages of high-flux hemodialysis (HD) in treating patients with chronic kidney disease (CKD) stage 5 are related to the higher toxin removal (especially 'middle molecules'), including sodium and water, and to the better biocompatibility of the treatment, including membrane and water quality. Several observational studies have shown that high-flux HD has positive effects on the survival and morbidity of uremic patients when compared with low-flux HD. The primary analysis of the prospective randomized HEMO (Hemodialysis Outcomes) study showed that high-flux HD was associated with an 8% nonsignificant reduction of mortality in comparison with low-flux HD. However, a secondary analysis pointed to an advantage for high-flux HD in subgroups of patients. More recently, the MPO (Membrane Permeability Outcome) study found that survival could be significantly improved by using high-flux HD compared with low-flux HD in high-risk patients as identified by serum albumin <=4 g/dl and, in a post hoc analysis, in diabetic patients as a whole. On-line hemodiafiltration (HDF) is considered the most efficient technique of using high-flux membranes. Clearance of small solutes like urea are higher than in hemofiltration, and clearance of middle solutes like beta(2)-microglobulin are higher than in high-flux HD. Since there is only a very limited number of randomized prospective trials comparing HDF and high-flux HD, no conclusive data are available about the effect of increased convection of on line HDF on survival and morbidity of CKD patients. The suggested advantages of HDF must be confirmed by a large randomized controlled study. PMID- 21625097 TI - Antioxidant dialytic approach with vitamin E-coated membranes. AB - Oxidative stress is prevalent in dialysis patients and has been implicated in the pathogenesis of anemia and cardiovascular disease. Vitamin E-coated membranes are low-flux dialyzers consisting of a multilayer membrane with liposoluble vitamin E on the blood surface allowing direct contact with free oxygen radicals to be scavenged on the membrane site. The antioxidant properties of these membranes have an important clinical benefit because of reducing oxygen stress and inflammation may contribute to an improvement of hemoglobin levels, lower recombinant human erythropoietin dose and better anemia management, and at the same time may have a favorable impact on cardiovascular complications. PMID- 21625098 TI - Dynamic hemodialysis: a potential solution for middle molecule removal. AB - Better clearance of middle molecular weight solutes has been associated with improved patient outcomes. However, blood-membrane interaction during dialysis results in the development of secondary protein layers on the dialysis membrane surface, leading to impairments in middle molecular weight solute removal. The shear rate at the surface of the dialysis membrane is one of the important factors that influence blood-membrane surface interaction. In this paper, we discuss the concept of 'dynamic hemodialysis', i.e. using mechanical vibration to increase shear rate at the surface of dialysis membrane. For longitudinal and transverse vibratory motions, hemodynamic changes inside the membrane and their consequent effects on membrane morphology and dialysis efficiency are discussed. Longitudinal vibration generates a reverse flow by relative membrane motion, and transverse vibration generates a symmetric swirling flow inside the hollow fiber, which increases shear rate and enhances flow mixing. Additionally, the impulse induced by the vibration could sustain the absorption capacity of the membrane to specific uremic toxins. In conclusion, dynamic hemodialysis could enhance solute removal by minimizing the morphological changes in the dialysis membrane during hemodialysis. However, the operating conditions of dynamic hemodialysis (i.e. the amplitude and frequency of mechanical vibration) need to be optimized in terms of hemolysis and noise from the mechanical vibration. PMID- 21625099 TI - Metabolic derangements seen in chronic kidney disease and end-stage renal disease patients. AB - The National Health and Nutrition Examination Survey (NHANES) shows increasing trends in the prevalence of chronic kidney disease (CKD) and end-stage renal disease (ESRD) in the US population. There is an inverse relationship between a declining glomerular filtration rate and mortality and morbidity. A wide spectrum of complex metabolic derangements contributes to a large extent for this increased mortality and morbidity. An overview of some of these metabolic derangements--like uremic dyslipidemia, uremic inflammation, and endocrine abnormalities affecting thyroid and sexual functioning of CKD patients--is provided. Understanding some of these metabolic derangements may help develop new and affective strategies to help reduce mortality and morbidity among CKD patients. PMID- 21625100 TI - Recent insights in inflammation-associated wasting in patients with chronic kidney disease. AB - In patients with end-stage renal disease (ESRD), inflammation, and protein energy wasting (PEW) are two highly prevalent and interconnected entities, jointly exerting a deleterious effect on multiple other ESRD-specific pathological processes and eventually on patient outcome. With respect to the pathophysiology underlying this strong association, knowledge has been actively expanded over the past few years. As such, it is nowadays recognized that inflammation acts via direct, as well as indirect, pathways in its contribution to PEW. Directly, inflammation causes alterations in amino acid utilization, translating into increased catabolism and decreased anabolism of muscle tissue. Indirectly, inflammation may act via altered ghrelin and adipokine metabolism, adipose tissue distribution, and pathological neuroendocrine signaling, as well as coexistent depression in inducing anorexia and PEW. In addition, two relatively new inflammatory markers (pentraxin-3 and TNF-like weak inducer of apoptosis) have gained attention with respect to their roles in this specific context. The current review deals with recent updates in the literature on the aforementioned pathways connecting inflammation to PEW and subsequent mortality. PMID- 21625101 TI - Factors determining insulin resistance in chronic hemodialysis patients. AB - Insulin resistance (IR) is common in chronic hemodialysis (CHD) patients and is associated with excess mortality. The gold standard for assessment of insulin sensitivity is hyperinsulinemic euglycemic clamp studies which provide the precision and accuracy necessary, especially for mechanistic studies. However, clamp studies are labor-intensive and complicated for more practical use. Accordingly, additional indices such as homeostatic model assessment of insulin resistance (HOMA), quantitative insulin sensitivity check index, and adipokine based measurements represent appropriate alternatives for large epidemiological and interventional studies. The etiology of IR in the CHD population is complex and multifactorial. The predominant pathophysiological mechanism of 'uremic insulin resistance' is a post-receptor defect in the skeletal muscle; however, other glucose metabolism abnormalities are also present. Some of the proposed determinants of IR in CHD patients include chronic inflammation, excess visceral fat, adipokine deregulation and accumulation, metabolic acidosis, oxidative stress, vitamin D deficiency, anemia, decreased physical activity, and accumulation of uremic toxins. The relative importance of each of these abnormalities is not well-defined, although excess visceral fat and inflammation seem to be the most important correlates of IR in this patient population. There are only few interventional studies targeted at improving insulin resistance in CHD patients. Insulin sensitizers such as metformin and PPAR-gamma agonists are either contraindicated or sparingly used due to their potential side effects, even in CHD patients with overt diabetes mellitus. More novel approaches to improving IR in this patient population might lead to potential strategies for preventing excess mortality. PMID- 21625102 TI - Lipid abnormalities in patients with chronic kidney disease. AB - Cardiovascular disease is increased in patients with chronic kidney disease (CKD) and is the principle cause of morbidity and mortality in these patients. Dyslipidemia, while common in these patients, is usually not characterized by elevated cholesterol, except in those patients with massive proteinuria. Qualitatively, increased triglycerides and reduced high-density lipoproteins (HDL) are most frequently described. Extensive abnormalities in the metabolism of apolipoprotein (apo) B-containing lipoproteins have been demonstrated, including those derived from the gut (apoB-48) as well as those derived from hepatic synthesis (apoB-100). Decreased enzymatic delipidation, in addition to reduced receptor removal of these lipoproteins, results in increased concentrations of these apoB-containing moieties, and in particular, their atherogenic remnants. Abnormalities in apoA-containing lipoproteins are also present and these changes may contribute not only to the lower levels of HDL seen, but also to the proinflammatory state that is frequently present in CKD patients. As a result, therapeutic strategies designed to modify atherosclerotic-caused outcomes in CKD may require multiple approaches. PMID- 21625103 TI - Statins and lipid-lowering strategies in cardiorenal patients. AB - The prevalence of chronic kidney disease (CKD) is increasing alarmingly mainly as a result of an ongoing epidemic of obesity, metabolic syndrome, and diabetes mellitus. CKD is a well-recognized risk multiplier for development of cardiovascular disease (CVD), and it is widely known that CVD is the leading cause of morbidity and mortality in patients with CKD. Cardiovascular (CV) morbidity and mortality is significantly increased along the continuum of CKD, and it is more than 10 times higher in end-stage renal disease populations than in the general population. Lipid metabolism is profoundly disturbed in CKD, and there is a gradual shift to the uremic lipid profile as kidney function deteriorates, which is further modified by the presence of comorbidities such as diabetes and obesity. Apart from quantitative differences, major qualitative changes in lipoproteins can be observed, such as oxidization and modification to small and dense low-density lipoprotein (LDL), which render the particles more atherogenic. It has been noted that these abnormalities contribute to the development of CV events, and they may lead to the progression of CKD. Lipid lowering treatment with statins in the general population has achieved important benefits both in reducing CV risk and in the prevention of CVD. Similarly, data from secondary analyses of CKD subgroups of larger prospective trials using statins also suggest an important benefit on CV outcomes and, with more conflicting evidence, on the progression of kidney disease. Preliminary results from a large randomized controlled trial of lipid-lowering therapy in CKD confirm similar benefits of treatment for dyslipidemia in patients with CKD and ESRD. The safety profile of lipid- lowering therapy with statins in CKD is not different from that observed in people with normal renal function. Hence, lipid- lowering therapy with statins should be part of the standard treatment of patients with CKD. PMID- 21625104 TI - Restoring the physiology of vitamin D receptor activation and the concept of selectivity. AB - Abnormalities of bone mineral parameters (calcium, phosphate, vitamin D, and parathyroid hormone) are nearly always present in patients with chronic kidney disease (CKD). These typically consist of hypocalcemia, hyperphosphatemia, abnormalities of vitamin D metabolism, and secondary hyperparathyroidism, and are now defined as CKD mineral bone disorders (CKD-MBD). Currently, emerging evidence indicates that deficiencies in vitamin D receptor (VDR) activation play crucial roles in adversely affecting the cardiovascular health of CKD patients. VDRs are not restricted to skeletal tissue, but are instead widely expressed throughout the body at several sites, such as in cardiac tissue, vascular smooth muscle cells, endothelial cells, renal tissue, and cells of the immune system. Restoring the physiology and modulation of VDR activator levels results in correlative regulatory effects on mineral homeostasis, hypertension, cardiovascular disease, and vascular calcification, as well as a number of other endpoints in cardiac and renal pathology. Among the compounds available for treatment of CKD-MBD, paricalcitol is a selective VDR activator. The term 'selective' refers to paricalcitol being more selective in affecting VDR pathways in the parathyroid gland compared with bone and intestine. As such, paricalcitol's selectivity allows for a wider therapeutic window with effects beyond parathyroid hormone control and mineral management, and may explain, in part, the increased survival advantage with paricalcitol treatment. PMID- 21625105 TI - Unmet need in renal protection--do we need a more comprehensive approach? AB - Progressive renal function loss is still a threat to health despite the presence and use of renal protective drugs such as ACEi and ARB. This unmet need appears to be related to factors like a residual high blood pressure and residual albuminuria. Strategies that enhance the efficacy of RAAS blockade, such as increasing the dose of ACEi or ARB, combining ACEi and ARB, or new RAAS inhibitor drugs like the direct renin inhibitors may help us in improving renal protection. However, it is likely that renal disease progression is caused by multiple risk factors and that we will need multiple approaches to tackle this residual risk. Recent trials testing the effect of vitamin D receptor activators and the effect of statins indicate that drugs in these classes may become valuable extra tools (in addition to ACEi or ARB) to tackle the unmet need in renal protection. PMID- 21625106 TI - The impact of paricalcitol on left ventricular hypertrophy. AB - Cardiovascular (CV) morbidity and mortality are significantly higher in patients with chronic kidney disease (CKD). Mineral metabolism disorders, such as hyperphosphatemia, hypocalcemia, and vitamin D deficiency, have been deeply associated not only with bone disease, but also with vascular calcification and CV disease. In addition, the decrease in vitamin D production stimulates the renin-angiotensin-aldosterone system, resulting in vasoconstriction and salt and water retention, which further promotes arterial stiffening. Several studies have shown that supplementation with vitamin D ameliorates some of these issues and is associated with improved survival. However, vitamin D also elevates serum levels of calcium and phosphorus. Selective vitamin D receptor (VDR) activators, such as paricalcitol, provide similar efficacy but are not associated with elevated serum concentrations of calcium and phosphorus. By selectively activating VDR, paricalcitol should enhance cardiorenal protection and provide significant clinical benefit. Therefore, paricalcitol may offer a novel and interesting approach to supplement and potentially enhance the standard of care in CKD patients. PMID- 21625107 TI - Vitamin D receptor activation: clinical outcomes. AB - Vitamin D has been recognized for years as a key player in the control of bone metabolism through the regulation of calcium and phosphate homeostasis. Vitamin D deficiency appears to be common in the general population as a result of decreased dietary intake, diminished absorption, or limited exposure to sunlight. Vitamin D deficiency results in increased circulating levels of parathyroid hormone, which has several disadvantageous effects on bone metabolism. The intake of ordinary doses of vitamin D supplements appears to be associated with a decrease in total mortality rates. In the chronic kidney disease (CKD) population, vitamin D receptor (VDR) activation deficiency is even more severe than in a non-CKD population, with a 25-(OH) vitamin D level being an independent predictor of all-cause mortality. Similarly to nonrenal populations, the corrections of vitamin D deficiency/treatment with vitamin D is associated with better survival. Selective VDR activators appears to be better than calcitriol in terms of survival or adverse effects. PMID- 21625108 TI - Vitamin D receptor activation: implications for daily practice. AB - Many clinical studies have focused on the relationship between outcome events and serum 25-OH vitamin D levels, despite the fact that most effects are mediated by vitamin D receptor (VDR) activation, the most proximate ligand for which is 1,25 OH(2) vitamin D. Evaluation of the separate components of this axis are well described, but an integrated view of the components, and the distinctions between the effects and consequences of substrate deficiency (25-OH vitamin D), and the direct effects of VDR activation, is the center of focus and ultimately impacts on the clinical approach to patents with chronic kidney disease (CKD). The focus of this contribution is on the specially defined need for vitamin D treatment in patients with CKD, and the importance of VDR activation in CKD, especially in patients with proteinuria. With the addition of the fibroblast growth factor-23 loop, and the realization that albuminuria represents several critical integrated levels of renal function, there is an emerging emphasis on the proximal tubule, interstitial inflammation and fibrosis. These interacting factors reflect fundamental mechanisms that culminate in progressive loss of kidney function(s), with loss of the glomerular filtration rate the most prominent feature. New therapeutic approaches could potentially intervene along these pathways, but they will have to be rigorously evaluated with defined outcome studies and cost benefit ratio analyses. The ultimate impact of chronic renal replacement therapy and cardiorenal death are the measures by which all such innovations must be judged. PMID- 21625109 TI - Body composition and heart rate variability to achieve dry weight and tolerance. AB - Autonomic dysfunction in patients with end- stage renal disease is associated with poor prognosis. Heart rate variability (HRV), determined by the standard deviation of the normal R- R interval, has been reported to be a useful evaluation of cardiac autonomic modulation. The relationship between HRV and hydration status (HS) can be analyzed by whole body bioimpedance spectroscopy. This allows a classification of patients according the combination of HS with predialysis systolic blood pressure. Differences in HRV can be studied in patients with high over hydration, but normal or low blood pressure, with respect to fluid-overloaded/hypertensive patients and normohydrated/normotensive patients. In conclusion, the assessment of the autonomic nervous system response to the hemodialysis treatment in end- stage renal disease patients, classified according to a reliable and quantitative measurement of their fluid overload, could permit better management of both arterial blood pressure and HS. PMID- 21625110 TI - Noninvasive volume assessment in the emergency department: a look at B-type natriuretic peptide and bioimpedance vector analysis. AB - B-type natriuretic peptide (BNP) and bioimpedance vector analysis (BIVA) are potential methods of assessing fluid status in patients. Their speed and ease of implementation allows them to be used at the point of care. Currently, BNP is pivotal in the diagnosis and prognosis of heart failure, and is starting to be implemented in other diseases (e.g. acute coronary syndrome and end-stage renal disease). Although it can be elevated from volume-overload-induced cardiac stress, when assessing volume this method is ultimately unreliable since it lacks adequate specificity and accuracy. Alternatively, BIVA is accurate and sensitive in the detection of fluid status, but cannot be used to tell the etiology of the volume aberration. Despite being relatively new, BIVA is becoming recognized as a superior method to assess volume. PMID- 21625111 TI - Continuous real-time urine output monitoring for early detection of acute kidney injury. AB - In critically ill patients, acute kidney injury (AKI) is a common complication. In some cases, oliguria may be the only sign verifying this condition. The consensus definitions of RIFLE and AKIN are based on changes in creatinine and urine output and define classes of severity within AKI. While meaningful change in serum creatinine is often not detectable until 48 h after deterioration in kidney function, urine output is a more rapid physiological parameter and detectable at the patient's bedside. Although urine output is a critical parameter in the intensive care unit, routine urine output measurements are performed manually. As a result, they may not be done timely and may be subject to inaccuracies due to human factors. The URINFO((r)) system is an innovative digital urine meter that provides continuous minute-to-minute monitoring of urine output, thereby enhancing kidney monitoring and the acquisition of more reliable urine output information in realtime. Consequently, monitoring of urine output with URINFO may enable rapid therapeutic interventions and can be incorporated into patient data systems, thereby improving therapy management. PMID- 21625112 TI - Extracorporeal ultrafiltration for congestive heart failure patients. AB - In the US and Europe, approximately 90% of heart failure (HF) hospitalizations are due to symptoms and signs of sodium and fluid excess. Congestion is associated with HF progression. According to data from large national registries, approximately 40% of hospitalized HF patients are discharged with unresolved congestion, which may contribute to unacceptably high rehospitalization rates. Diuretics reduce the symptoms and signs of fluid overload, but their effectiveness can be reduced by excess salt intake, underlying chronic kidney disease, renal adaptation to their action, and neurohormonal activation. In addition, the production of hypotonic urine limits the ability of loop diuretics to reduce total body sodium. Ultrafiltration is the mechanical removal of fluid from the vasculature. Clinical studies of ultrafiltration have shown that removal of isotonic fluid relieves symptoms of congestion, improves cardiac filling pressures and exercise capacity, and restores diuretic responsiveness in patients with diuretic resistance, concomitantly with favorable effects on pulmonary function, ventilatory efficiency, and neurohormonal activation. Ultrafiltration has been shown to reduce rehospitalizations in a randomized controlled trial of patients with decompensated HF. Future larger controlled clinical trials should evaluate the effect of ultrafiltration on survival. PMID- 21625113 TI - Acute kidney injury biomarkers and endpoints for clinical trials. AB - Acute kidney injury (AKI) is associated with a heavy burden of morbidity and mortality, despite advances in intensive care and the management of high-risk patients. Numerous clinical trials have failed to ameliorate the outcomes of AKI. The proposal and validation of standardized definitions and staging criteria for AKI has been an important step in improving the conduct of clinical trials in this population. Similarly, a multidisciplinary dialogue is making progress towards standardization of the clinical trial endpoints to prove efficacy and effectiveness in AKI research. Taken together with the increasing availability of timely, sensitive, and specific novel biomarkers of kidney damage, we are poised to use these tools to conduct successful clinical trials of agents for the prevention and treatment of this devastating clinical syndrome. PMID- 21625114 TI - Impact of acute kidney injury on chronic kidney disease and its progression. AB - Acute kidney injury (AKI) is a devastating clinical problem that affects a growing number of patients, especially elderly ones, and is associated with high morbidity and mortality. It was previously thought that patients who survive an episode of AKI recover renal function without further sequelae; however, recent population- based studies suggest that this may not be the case. New clinical studies suggest that a strikingly large percentage of patients who have AKI do not fully recover renal function or require permanent renal replacement therapy, and that this population has an important impact on the epidemiology of chronic kidney disease (CKD) and end-stage renal disease. These clinical studies verify animal studies that have established a link between AKI and CKD progression. Future clinical studies are underway to prospectively characterize the natural history of AKI and CKD progression and to identify predictive biomarkers. PMID- 21625115 TI - Acute kidney injury and its management. AB - Acute kidney injury (AKI) is a life-threatening disorder, but one which is potentially reversible. This syndrome is a frequent and serious complication of hospitalized patients. When severe enough to require renal replacement therapy, hospital mortality approaches 60% and recovery among survivors may be as poor as 50%. Moreover, recent evidence using uniform definitions and classifications have revealed that even less severe forms of AKI are associated with reduced survival and other long-term adverse consequences, including progression of chronic kidney disease. Promising new biomarkers are becoming available and new strategies for prevention of AKI in specific situations are being developed. Advanced treatment options, including adsorptive therapy, the renal tubular assist device and stem cell therapy, are also on the horizon. Increasing knowledge in this field is beginning to fill in the missing jigsaw puzzle pieces and a more coherent picture is emerging. Challenges in management of AKI to reduce mortality are sill daunting, however, and more research is urgently needed. The combination of education, risk stratification, prevention, early detection, prompt therapeutic intervention, quality of supportive care, and innovative therapies offer the promise of improving outcomes in patients afflicted with this serious condition. PMID- 21625116 TI - Wearable dialysis: what is missing? AB - In this chapter, unanswered questions regarding the safety and usability of the wearable artificial kidney (WAK) are discussed. This will cover issues such as safe blood access, risks related to the operation of the WAK, temporary disconnection or separation during sleep, and reloading dialysate regeneration cartridges. PMID- 21625117 TI - Current technological approaches for a wearable artificial kidney. AB - A wearable artificial kidney (WAK) is a device that continuously supports renal function during ambulation or social activities out of hospital. With the aim of improving dialysis patients' quality of life, WAK systems have been in development for several decades. Technological evolutions in dialysis membrane and dialysate regeneration have been paving the way to wearability, and the possibility of implantation, for renal replacement therapies. However, at present, there are many technical issues confronting the attempts to apply WAK systems in clinical practice. Here, we have reviewed the necessary technical requirements and the WAKs currently being developed that are trying to meet these. Aside from technical issues, ethical, legal and economic aspects should be also considered together, in order to minimize trial and error in the development of the WAK. Continuous follow- up, integration with emerging new technologies, and multidisciplinary approaches involving clinicians, engineers, economists and social scientists are required for the realization of WAK in clinical practice. PMID- 21625118 TI - From wearable ultrafiltration device to wearable artificial kidney. AB - INTRODUCTION: Since the inception of hemodialysis as a treatment for patients with chronic kidney disease, there have been designs created for portable, wearable or implantable devices to improve both quality of therapy delivered and patient quality of life. METHODS: We conducted a pilot proof- of- concept study in 8 stable adult hemodialysis patients using a wearable hemodialysis device with a sorbent-based regeneration dialysate system. RESULTS: All patients were safely treated for 4-8 h. Ultrafiltration was successfully performed without any cardiovascular effects, with a reduction in extracellular fluid to total body fluid ratio (from 0.339 +/- 0.003 to 0.335 +/- 0.003 after treatment; p=0.002). Respective clearances (ml/min) were: 22.6 +/- 1.8 for urea, 20.7 +/- 1.8 for creatinine, 21.7 +/- 1.8 for phosphate and 11.6 +/- 1.8 for beta(2) microglobulin. Although the small solute clearances are much lower than for conventional intermittent hemodialysis, this device has been designed to operate for sustained periods, and therefore would be predicted to have equivalent small solute clearances to continuous dialysis treatments (CRRT) in the intensive setting. Safety mechanisms were shown to operate promptly in a case of a venous needle disconnection, and in cases of circuit clotting. CONCLUSION: This was a pilot study designed to demonstrate the safety of a wearable hemodialysis device. In addition to confirming safety, we were able to confirm that the device not only had similar clearances to CRRT used in the intensive-care setting, but also increased clearances of middle molecules, such as beta(2)-microglobulin and phosphate. PMID- 21625119 TI - Remote monitoring for the wearable artificial kidney. AB - Recent technological advances have led to the construction of a truly wearable artificial kidney (WAK). This device offers the promise of regulation of volume status and provision of adequate solute clearances, while at the same time allowing its user to receive continuous therapy while going on with the normal activities of daily life. An important component of such a therapy is the ability to remotely monitor the treatment. This remote monitoring allows assurance of patient safety through surveillance of critical portions of the treatment as well as the ability to monitor compliance with prescribed therapy and automated collection of treatment data. Recent advances in telemedicine, telemonitoring, remote network access and sensor technologies have made such remote monitoring of the WAK feasible. Furthermore, experience with remote monitoring for nocturnal hemodialysis, which shares some similarities with the needs of the WAK for monitoring, can guide implementation of such a system. PMID- 21625120 TI - Anemia trials in CKD and clinical practice: refining the approach to erythropoiesis-stimulating agents. AB - As our knowledge of managing renal anemia with recombinant human erythropoietin has grown, new clinical challenges have emerged. Although initial observational studies produced important favorable results, troubling observations later came from 4 randomized controlled trials that assigned CKD patients to intervention with an erythropoiesis-stimulating agent (ESA) to achieve high hemoglobin levels (Hb). The difference between the observational studies and these randomized controlled trials comes from the observation of higher cardiovascular events when targeting higher Hb, but lower events once patients had achieved these Hb targets. In all of the 4 RCTs, the primary intervention was 'targeting' a higher Hb, and using higher dosages of ESA as the means. Therefore, the question to be solved is: what is harmful with recombinant human erythropoietin--targeting higher hemoglobin or achieving higher hemoglobin? Evidence supporting a relationship between ESA exposure and worse outcomes is not universal, and these comorbidities would require the administration of high doses. However, increasing ESA doses in hyporesponsive patients to achieve a specific target is not reasonable. Managing anemia in CKD patients is complex. It is affected by the underlying disease, comorbid conditions, the environment and several other factors that differ among patients. Thus, anemia management in these patients needs an individualized approach. Each patient should be treated according to a Hb target with the lowest effective ESA dose, while avoiding large fluctuations in Hb levels or prolonged periods outside the target. This strategy may necessitate changes to the ESA dose, dosing frequency and iron supplementation over the course of a patient's treatment, and proactive management of conditions that can affect ESA responsiveness. While all ESAs effectively increase Hb levels, differences with respect to route of administration, pharmacokinetics, and dosing frequency and efficiency should be considered to maximize the benefits of ESA treatment for the individual patient. PMID- 21625121 TI - New erythropoiesis-stimulating agents and new iron formulations. AB - Today, erythropoiesis-stimulating agents (ESAs), together with iron supplementation, are the main tool for anemia correction in chronic kidney disease patients. Over the past decades, a number of attempts have been made to modify the erythropoietin molecule in order to improve its pharmacokinetic and pharmacodynamic properties. More recently, small peptides, which are unrelated to erythropoietin but bind to the same receptor, have been developed. In addition to this, other strategies to stimulate erythropoiesis have been followed, such as activin inhibition or stabilization of hypoxia-inducible transcription factors. Interestingly, the latter have the advantage of being administered orally. New iron molecules, such as ferumoxytol, ferric carboxymaltose and iron isomaltoside 1000, have recently been marketed. These new agents can administered at high doses while releasing minimal free iron. Their safety profile is good, but long- term post- marketing data are still needed to evaluate the occurrence of rare adverse events. PMID- 21625122 TI - Impact of dialysis technique on renal anemia. AB - Cardiovascular disease is a significant complication in chronic kidney disease (CKD) and a major cause of death in dialysis patients. Clinical studies have shown that anemia is associated with reduced survival in patients undergoing chronic hemodialysis. Furthermore, an association between anemia and adverse cardiovascular outcomes has also been observed in patients with earlier stages of CKD not yet requiring dialysis. Although this fact still remains controversial, high-efficiency on-line hemodiafiltration (HDF) has been shown to improve anemia and to reduce the need for erythropoietin-stimulating agents in hemodialysis (HD) patients. This positive effect has been attributed to the fact that the convective methods might remove some protein-bound erythropoietic inhibitor substances. Moreover, in HD patients, renal anemia is linked to the inflammatory state of uremic syndrome. It is also worth nothing that the improvement in anemia is associated with a reduced inflammatory state in patients undergoing on-line HDF. Here, we have reviewed the current knowledge of the effect of dialysis technique on renal anemia. PMID- 21625123 TI - Cardio-renal anemia syndrome. AB - Heart failure (HF) is a systemic disease that also involves organs and tissues other than the heart and the vasculature. Between 25 and 50% of patients with HF are also affected by some degree of kidney disease. Anemia may be present in patients with HF, particularly if the kidney is also affected. Anemia is observed in about 20% of patients with ambulatory HF, but its prevalence may increase to 60% or more in patients with advanced disease or significant co-morbidities. Cardio-renal anemia syndrome (CRAS) represents a pathological triangle in which the primary failing organ is either the heart or the kidney, and the dysfunction of one leads to dysfunction of the other. Mortality rates increase with only anemia or kidney disease being present in patients with HF. The full clinical picture of CRAS is present in about 20% of all patients hospitalized for HF. In such patients, a steep increase in mortality rates has been observed. This article describes the suggested classification systems of CRAS, its clinical significance, and potential therapeutic avenues. PMID- 21625124 TI - Transition m.3308T>C in the ND1 gene is associated with left ventricular hypertrabeculation/noncompaction. AB - Though left ventricular hypertrabeculation/noncompaction (LVNC) is frequently associated with mitochondrial DNA (mtDNA) mutations, it has not been reported in association with the transition m.3308T>C of the NADH dehydrogenase subunit 1 (ND1) gene. The index patient is a 16-year-old Tunisian female who was investigated for a systolic murmur and cardiomegaly. Echocardiography revealed tricuspid insufficiency, moderate left ventricular dilatation, Ebstein's anomaly, a superior caval vein draining into the coronary sinus and, surprisingly, LVNC of the apex and the lateral wall. LVNC was absent in all other cardiologically investigated siblings. RNA and mtDNA sequence analysis revealed the known homoplasmic mutation m.3308T>C resulting in the replacement of the first amino acid methionine by threonine in the ND1 subunit of respiratory chain complex I. The m.3308T>C mutation was also present in the patient's mother and several other family members but absent in 350 controls. Additionally, the index patient carried the polymorphisms m.8248A>G in the COX2 gene and m.8468C>T in the ATP8 gene. It is concluded that LVNC may be associated with the known homoplasmic m.3308T>C mutation in the ND1 gene. However, the pathogenetic role of this mutation in the development of LVNC remains elusive. PMID- 21625125 TI - Validity of an information and communication technology system for data capture in epidemiological studies. AB - The Belgian National Institute of Health Insurance is implementing an oral health data registration and surveillance system. This study aimed to develop and validate a system of electronic data capture for oral health surveys at a national level - Oral Survey-B - and to identify the advantages and disadvantages of the electronic system in comparison with the traditional handwritten data capture. Six series of full-mouth recordings simulating the clinical examination of 6 patients were set up in a Powerpoint presentation. The validation was undertaken by 52 general practitioners. A randomized one-period crossover design was used with two formats of data capture, i.e. electronic followed by handwritten or handwritten followed by electronic system. Further, 6 benchmarked handwritten forms were transferred to the electronic format. For the electronic data capture, 86.5% of the practitioners had a correct completion rate of >=95%. The corresponding value for the handwritten data capture and transfer was 78.8% (p = 0.25, McNemar test). The overall accuracy of forms without any error was 73.4% for the electronic and 62.5% for the handwritten data capture (p < 0.001, signed-rank test). Significantly lower percentages of errors and less time were observed for the electronic data capture (p < 0.001, signed-rank test). Practitioners considered the electronic data capture as being much more difficult to carry out (p < 0.001). As information technology has turned into an ever more necessary working tool in epidemiology, there should be an important potential for uptake of further improvements in electronic data capture in the future. PMID- 21625126 TI - Clinical performance of two fluorescence-based methods in detecting occlusal caries lesions in primary teeth. AB - This in vivo study aimed to evaluate the performance of 2 fluorescence-based methods in detecting occlusal caries lesions in primary teeth, compared with the performance of visual inspection and radiographic methods, and to propose a mathematic correction of the diagnostic parameters due to the imperfect reference standard method used in the study. Two examiners assessed the occlusal surfaces of 407 primary teeth (62 children) using visual inspection (ICDAS), radiographic, DIAGNOdent pen (pen type laser fluorescence; LFpen), and fluorescence camera (FC) methods. At the noncavitated threshold (NC) the reference standard method was the results of ICDAS, and at the dentine caries threshold (D3) teeth diagnosed with dentine caries by ICDAS or radiographic methods were subjected to operative treatment to confirm the presence of lesion. Reproducibility, sensitivity, specificity, accuracy, and the area under the ROC curve were calculated for the methods at both thresholds. At the NC threshold, LFpen had a slightly better performance compared to the FC and radiographic methods. However, at the D3 threshold, both fluorescence-based methods performed similarly. Visual inspection and radiographic methods presented higher specificities but lower sensitivities than fluorescence methods. After corrections, there was a significant decrease in some parameters. In conclusion, both fluorescence-based methods presented similar performance in detecting occlusal dentine caries lesions in primary teeth, but they usually gave more false-positive results than did the visual and radiographic methods. The correction proposed shows that the performance of the methods can be overestimated, and the correction should be validated and considered in further studies that use an imprecise reference standard method. PMID- 21625127 TI - Methodology and models in erosion research. Foreword. PMID- 21625128 TI - Dental erosion--an overview with emphasis on chemical and histopathological aspects. AB - The quality of dental care and modern achievements in dental science depend strongly on understanding the properties of teeth and the basic principles and mechanisms involved in their interaction with surrounding media. Erosion is a disorder to which such properties as structural features of tooth, physiological properties of saliva, and extrinsic and intrinsic acidic sources and habits contribute, and all must be carefully considered. The degree of saturation in the surrounding solution, which is determined by pH and calcium and phosphate concentrations, is the driving force for dissolution of dental hard tissue. In relation to caries, with the calcium and phosphate concentrations in plaque fluid, the 'critical pH' below which enamel dissolves is about 5.5. For erosion, the critical pH is lower in products (e.g. yoghurt) containing more calcium and phosphate than plaque fluid and higher when the concentrations are lower. Dental erosion starts by initial softening of the enamel surface followed by loss of volume with a softened layer persisting at the surface of the remaining tissue. Dentine erosion is not clearly understood, so further in vivo studies, including histopathological aspects, are needed. Clinical reports show that exposure to acids combined with an insufficient salivary flow rate results in enhanced dissolution. The effects of these and other interactions result in a permanent ion/substance exchange and reorganisation within the tooth material or at its interface, thus altering its strength and structure. The rate and severity of erosion are determined by the susceptibility of the dental tissues towards dissolution. Because enamel contains less soluble mineral than dentine, it tends to erode more slowly. The chemical mechanisms of erosion are also summarised in this review. Special attention is given to the microscopic and macroscopic histopathology of erosion. PMID- 21625129 TI - Methods for the measurement and characterization of erosion in enamel and dentine. AB - The advantages, limitations and potential applications of available methods for studying erosion of enamel and dentine are reviewed. Special emphasis is placed on the influence of histological differences between the dental hard tissue and the stage of the erosive lesion. No method is suitable for all stages of the lesion. Factors determining the applicability of the methods are: surface condition of the specimen, type of the experimental model, nature of the lesion, need for longitudinal measurements and type of outcome. The most suitable and most widely used methods are: chemical analyses of mineral release and enamel surface hardness for early erosion, and surface profilometry and microradiography for advanced erosion. Morphological changes in eroded dental tissue have usually been characterised by scanning electron microscopy. Novel methods have also been used, but little is known of their potential and limitations. Therefore, there is a need for their further development, evaluation, consolidation and, in particular, validation. PMID- 21625130 TI - Screening and prediction of erosive potential. AB - The literature on the erosive potential of drinks and other products is summarised, and aspects of the conduct of screening tests as well as possible correlations of the erosive potential with various solution parameters are discussed. The solution parameters that have been suggested as important include pH, acid concentration (with respect to buffer capacity and concentration of undissociated acid), degree of saturation, calcium and phosphate concentrations, and inhibitors of erosion. Based on the available data, it is concluded that the dominant factor in erosion is pH. The effect of buffer capacity seems to be pH dependent. The degree of saturation probably has a non-linear relationship with erosion. While calcium at elevated concentrations is known to reduce erosion effectively, it is not known whether it is important at naturally occurring concentrations. Fluoride at naturally occurring concentrations is inversely correlated with erosive potential, but phosphate is probably not. Natural plant gums, notably pectin, do not inhibit erosion, so they are unlikely to interfere with the prediction of erosive potential. The non-linearity of some solution factors and interactions with pH need to be taken into account when developing multivariate models for predicting the erosive potential of different solutions. Finally, the erosive potential of solutions towards enamel and dentine might differ. PMID- 21625131 TI - Initial erosion models. AB - Initial dental erosion - the very first tooth surface changes caused by short term acidic exposures before any tissue loss is observed - has been studied using in vitro and in situ/in vivo protocols. This paper describes the events that should be considered when modelling initial erosion, the available protocols and their strengths and limitations. In vitro initial erosion models can provide relevant information about the erosive outcome. However, the published studies vary considerably regarding the erosive parameters used, and few of these studies have been validated. On the other hand, relevant in situ/in vivo studies are available that have studied the effect of salivary pellicle and preventive treatments on initial erosion, as well as the changes in salivary pH following exposure to acid solutions. Guidelines for good methodology in modelling initial erosion are proposed. PMID- 21625132 TI - In vitro and in situ erosion models for evaluating tooth substance loss. AB - Because of the difficulties in measuring erosion in vivo, a number of in vitro and in situ models have been developed and validated. These models are flexible and informative, allowing single as well as multiple variables to be examined under specific conditions using accurate measurement techniques over defined timelines, thus yielding useful data without harmful effects on individuals. This information, together with clinical findings, is essential for clinicians advising susceptible patients appropriately regarding the management of their condition. Little guidance is available, however, on the standardisation of in vitro and in situ protocols for erosive tooth wear studies, so it is difficult to make meaningful comparisons between investigations as experimental variables differ widely from study to study. The aim of this review was to collate the available data on models designed to assess erosive challenges which are severe enough to cause tissue loss as opposed to just softening of the surface structure. The different types of models, with their merits and pitfalls, are documented. Test substrates, disinfecting regimens and ethical considerations are discussed. The aims of this paper are to give guidance to the researcher on evidence-based in vitro and in situ erosive tooth wear methodology and to suggest best practice given current knowledge. PMID- 21625133 TI - Design of erosion/abrasion studies--insights and rational concepts. AB - In vitro and in situ studies modelling the wear of dental hard tissues due to erosion and abrasion are characterised by a high variation in study designs and experimental parameters. Based on a summary of the existing protocols, the present review aimed to describe and discuss the parameters which must be carefully considered in erosion-abrasion research, especially when it is intended to simulate clinical conditions. Experimental characteristics and parameters were retrieved from a total of 42 in vitro and 20 in situ studies. The key experimental characteristics included parameters of erosion (duration and pH) and abrasion (duration, kinds of toothbrush and toothpaste, brushing force, and time point) as well as co-factors (e.g. dental hard tissue). The majority of studies used models with alternating erosion/abrasion treatments intended to simulate clinical conditions, while other studies exaggerated clinical conditions intentionally, often using only a single erosion/abrasion treatment. Both in vitro and in situ models shared a high level of standardisation, but several studies showed a trend to severe erosion (e.g. >5 min/cycle) or extensive brushing (e.g. >100 brushing strokes/cycle) at a high frequency and repetition rate. Thus, studies often tend to produce a higher amount of wear than in the clinical situation, especially as modifying biological factors (e.g. the dilution of the erosive solution by saliva and the protective effect of the pellicle) cannot be simulated adequately. With respect to the existing models, it seems advisable to diminish duration and frequency of erosion and abrasion to more realistic clinical conditions when the everyday situation is to be simulated. Experimental parameters must be chosen with care to ensure that the problem is investigated in an appropriate mode at standardised conditions and with adequate measuring systems to allow prediction of clinical outcomes. PMID- 21625134 TI - Clinical studies of dental erosion and erosive wear. AB - We define erosion as a partial demineralisation of enamel or dentine by intrinsic or extrinsic acids and erosive tooth wear as the accelerated loss of dental hard tissue through the combined effect of erosion and mechanical wear (abrasion and attrition) on the tooth surface. Most experts believe that during the last decade there has been a significant increase in the prevalence and severity of erosive tooth wear, particularly in adolescents. Even when erosive wear occurs in its milder forms, this is a matter of concern, as it may compromise the integrity of an otherwise healthy dentition in later life. The erosive wear process is complicated and modified by many chemical, behavioural and associated processes in the mouth. If interventions are to be developed it is therefore important that in vivo methods are developed to assess the outcomes of the erosion and erosive wear processes and the effects of interventions upon them. This paper discusses potential methods of investigating erosion and erosive wear in vivo and the difficulties associated with clinical studies. PMID- 21625135 TI - Methodology and models in erosion research: discussion and conclusions. AB - This paper summarises the discussions which took place at the Workshop on Methodology in Erosion Research in Zurich, 2010, and aims, where possible, to offer guidance for the development and application of both in vitro and in situ models for erosion research. The prospects for clinical trials are also discussed. All models in erosion research require a number of choices regarding experimental conditions, study design and measurement techniques, and these general aspects are discussed first. Among in vitro models, simple (single- or multiple-exposure) models can be used for screening products regarding their erosive potential, while more elaborate pH cycling models can be used to simulate erosion in vivo. However, in vitro models provide limited information on intra oral erosion. In situ models allow the effect of an erosive challenge to be evaluated under intra-oral conditions and are currently the method of choice for short-term testing of low-erosive products or preventive therapeutic products. In the future, clinical trials will allow longer-term testing. Possible methodologies for such trials are discussed. PMID- 21625136 TI - Effect of shunt operation on idiopathic normal pressure hydrocephalus patients in reducing caregiver burden: evidence from SINPHONI. AB - BACKGROUND/AIMS: Patients with idiopathic normal pressure hydrocephalus (iNPH) are often given shunt operations to reduce the triad symptoms (cognitive impairment, gait disturbance and urinary disturbance). We examined whether they also reduce caregiver burden. METHODS: The personal strain (PS) and role strain (RS) factors, which are related to the stress and constraints, respectively, on the caregivers of 81 iNPH patients were evaluated with the Zarit burden interview (ZBI) and each of the triad symptoms was evaluated with the iNPH grading scale (iNPHGS) before and 1 year after the shunt operation. RESULTS: Each of the iNPHGS scores, the total ZBI score and PS factor significantly improved after the shunt operation, but the RS factor did not. The improvement of cognitive impairment was the major factor in reducing caregiver burden. CONCLUSION: Shunt operations reduced the caregiver burden of iNPH patients. PMID- 21625138 TI - The missing link between obesity and hepatocellular carcinoma: IL-6-mediated STAT 3 activation as a key player in hepatocarcinogenesis. PMID- 21625137 TI - Visual rating versus volumetry to detect frontotemporal dementia. AB - BACKGROUND/AIMS: Automated, volumetrically defined atrophy in the left anterior cingulate (LAC) and anterior temporal regions (LAT) on MRI can be used to distinguish most patients with frontotemporal dementia (FTD) from controls. FTD and Alzheimer's disease (AD) can differ in the degree of anterior temporal atrophy. We explored whether clinicians can visually detect this atrophy pattern and whether they can use it to classify the 2 groups of dementia patients with the same accuracy. METHODS: Four neurologists rated atrophy in the LAC and LAT regions on MRI slices from 21 FTD, 21 controls, and 14 AD participants. Inter rater reliability and diagnostic accuracy were assessed. RESULTS: All 4 raters agreed on the presence of clinically significant atrophy, and their atrophy scoring correlated with the volumes, but without translation into high inter rater diagnostic agreement. CONCLUSIONS: Volumetric analyses are difficult to translate into routine clinical practice. PMID- 21625139 TI - Push-back technique facilitates ultra-low anterior resection without nerve injury in total mesorectal excision for rectal cancer. AB - AIMS: To describe our push-back approach to ultra-low anterior resection using the concept of the mucosal stump. METHODS: We mobilize the rectum using an abdominal approach, and perform mucosal cutting circumferentially at the dentate line. The mucosal stump is closed, and the internal sphincteric muscle resected partially or totally according to tumor location. Perianal dissection is performed along the medial plane of the external sphincteric muscles, and the hiatal ligament is dissected posteriorly. To resect the entire rectum, the closed rectal stump is pushed back to the abdominal cavity using composed gauze. This prevents injury to the autonomic nerve. RESULTS: We performed colonic J-pouch anal anastomosis using our mucosal stump approach in 58 patients with rectal cancer located <4 cm from the anal verge. According to the Wexner score, 7% of patients were fully continent, 71% had acceptable function with minor continence problems, and 22% were incontinent. No patients required intermittent self catheterization during follow-up. After a median follow-up of 49 months, there was only 1 case of local recurrence after surgery. CONCLUSION: Our push-back approach for internal sphincter resection produces satisfactory functional and oncological results in ultra-low anterior rectal cancer. PMID- 21625140 TI - European Association of Young Neurologists and Trainees: position paper on teaching courses for Generation Y. AB - The European Association of Young Neurologists and Trainees (EAYNT) is a non profit organization which acts on behalf of young neurologists in Europe and concertedly exerts influence on the formation of a new generation of neurologists [Struhal et al.: Eur J Neurol 2009;16:e146-e148]. This concerns particularly the Generation Y (Gen Y), also known as Millennial Generation, Digital Natives or Generation Next, a demographic cohort defined by birth between 1981 and 1999 [Elkind: Neurology 2009;72:657-663]. A unifying feature is the increased use and familiarity with online media and digital technologies. Online social networks and interactive communication have not only shaped this cohort but necessitate a different approach towards educational matters. This position paper aims to address the changing needs for Gen Yers in the context of education. PMID- 21625141 TI - Initial MRI findings predict progressive lacunar infarction in the territory of the lenticulostriate artery. AB - BACKGROUND/AIMS: The mechanisms underlying neurological deterioration in patients with acute lacunar infarction in the lenticulostriate artery (LSA) territory are currently unclear. We aimed to identify predictors for progressive neurological deficits using diffusion-weighted imaging (DWI). METHODS: We studied 40 consecutive patients who were classified into two groups based on their NIHSS scores: progressive infarction (PI) and non-progressive infarction (NPI). We calculated the size of DWI abnormalities and evaluated clinical characteristics on admission for both groups. RESULTS: 19 patients (47.5%) exhibited neurological deterioration. Time from onset to MRI was 8.2 +/- 6.7 h for all patients. DWI area was significantly larger in the PI relative to the NPI group (1.1 +/- 0.5 cm(2) for PI vs. 0.7 +/- 0.3 cm(2) for NPI; p = 0.002), although patients' NIHSS scores at admission were not significantly different between both groups. The optimal cut-off value of infarct area between PI and NPI was 0.98 cm(2). Multivariate analysis revealed that an infarct area on DWI >=0.98 cm(2) (odds ratio 10.57; 95% confidence interval 2.24-68.32; p = 0.006) was a significant independent predictor of PI. CONCLUSIONS: A large infarct area on initial DWI was an independent predictor of neurological deterioration in patients with acute lacunar infarctions in the LSA territory. PMID- 21625142 TI - An exploratory study on facial emotion recognition capacity in beginning Alzheimer's disease. AB - OBJECTIVE: It was the aim of this study to investigate facial emotion recognition (FER) in the elderly with cognitive impairment. METHOD: Twelve patients with Alzheimer's disease (AD) and 12 healthy control subjects were asked to name dynamic or static pictures of basic facial emotions using the Multimodal Emotion Recognition Test and to assess the degree of their difficulty in the recognition task, while their electrodermal conductance was registered as an unconscious processing measure. RESULTS: AD patients had lower objective recognition performances for disgust and fear, but only disgust was accompanied by decreased subjective FER in AD patients. The electrodermal response was similar in all groups. No significant effect of dynamic versus static emotion presentation on FER was found. CONCLUSION: Selective impairment in recognizing facial expressions of disgust and fear may indicate a nonlinear decline in FER capacity with increasing cognitive impairment and result from progressive though specific damage to neural structures engaged in emotional processing and facial emotion identification. Although our results suggest unchanged unconscious FER processing with increasing cognitive impairment, further investigations on unconscious FER and self-awareness of FER capacity in neurodegenerative disorders are required. PMID- 21625143 TI - Unusual dopaminergic depletion in variant Creutzfeldt-Jakob disease with early and rapid cognitive decline. PMID- 21625144 TI - The phonatory deviation diagram: a novel objective measurement of vocal function. AB - AIM: To identify the discriminative characteristics of the phonatory deviation diagram (PDD) in rough, breathy and tense voices. METHODS: One hundred and ninety six samples of normal and dysphonic voices from adults were submitted to perceptual auditory evaluation, focusing on the predominant vocal quality and the degree of deviation. Acoustic analysis was performed with the VoxMetria (CTS Informatica). RESULTS: Significant differences were observed between the dysphonic and normal groups (p < 0.001), and also between the breathy and rough samples (p = 0.044) and the breathy and tense samples (p < 0.001). All normal voices were positioned in the inferior left quadrant, 45% of the rough voices in the inferior right quadrant, 52.6% of the breathy voices in the superior right quadrant and 54.3% of the tense voices in the inferior left quadrant of the PDD. In the inferior left quadrant, 93.8% of voices with no deviation were located and 72.7% of voices with mild deviation; voices with moderate deviation were distributed in the inferior and superior right quadrants, the latter ones containing the most deviant voices and 80% of voices with severe deviation. CONCLUSION: The PDD was able to discriminate normal from dysphonic voices, and the distribution was related to the type and degree of voice alteration. PMID- 21625145 TI - Orangutans (Pongo pygmaeus) do not form expectations based on their partner's outcomes. AB - Several primate species form expectations based on others' outcomes, responding negatively when their outcomes differ from their partners'. The function and evolutionary pathway of this behavior are unknown, in part because all of the species which have been tested thus far share traits related to a gregarious lifestyle, intelligence, and cooperativeness. Our goal was to test whether inequity is a homology among primates or a convergence by comparing one species known to show social comparisons, the chimpanzee, to another great ape which differs on several of these life history characteristics. Using a protocol identical to one used previously with chimpanzees, we tested whether orangutans, an intelligent but predominantly solitary species with few opportunities to cooperate, responded similarly. To allow for a strong comparison with chimpanzees (and other species), we used socially housed adults of both sexes, tested with members of their social group. Orangutans did not respond negatively to inequity, supporting previous findings and indicating that inequity responses in apes are likely a convergence based on either sociality or cooperative tendency. These results in such closely related species highlight the need for additional comparative studies to understand better the function and evolution of social behaviors. PMID- 21625146 TI - Advances in the planning and delivery of radiotherapy: new expectations, new standards of care. AB - The practice of radiation therapy continues to build on rapid advancements in treatment planning and delivery technology, which brings real potential for improving treatment outcomes. Manufacturers have employed advanced computer and imaging technology to produce treatment planning/delivery systems capable of precise shaping of dose distributions, conformal target volume coverage for even the most complex shapes and conformal avoidance of specified sensitive normal structures. However, these new systems have led to a more complex, less intuitive planning and treatment delivery process that presents great challenges for quality assurance/treatment verification. Advances in planning and delivery technologies will continue to occur at record paces, pushing the field toward even higher expectations for radiotherapy accuracy, reliability and applicability and leading the field to new standards of care. However, this optimism must be tempered with the realizations that for this to happen, progress is urgently needed in 3 areas, (1) accuracy in specification of gross tumor volume and clinical target volume, (2) radiation oncology informatics and (3) quality assurance, if we are to keep pace with these rapid planning and delivery developments. PMID- 21625147 TI - Advanced technologies in the radiotherapy clinic: system fundamentals. AB - The radiotherapy treatment process is undergoing rapid development at every step from planning through delivery, and each step is increasingly automated and assisted by new imaging, positioning, contouring and treatment tools. Plan delivery and verification is now aided using an increasing range of image guidance technologies, and imaging at treatment now brings broad opportunities for dose guidance and adaptation for improving overall treatment quality. While these many tools bring exciting opportunities for exact, reliable and efficient targeting of radiation dose, a consistently high level of accuracy must be achieved at every step to achieve the desired results. This level of workmanship requires thorough understanding of the basic methods involved in each step, including the opportunities and limitations, by both the clinicians and the planning/delivery staff alike. These processes and their clinical implementation are discussed in depth throughout this volume. Here, we overview their integration and guiding background concepts, as well as a range of workday efficiencies for clinical practice. PMID- 21625148 TI - Controversies in the adoption of new healthcare technologies. AB - Healthcare economists generally agree that the development and rapid introduction of new technologies and the expanding utilization of existing ones in national healthcare systems have been significant factors in the dramatic and potentially unsustainable growth in healthcare spending. Creating a rational system for evaluation of emerging technologies in this country has been complicated by 3 broad issues: the often conflicting needs and expectations of the variety of stakeholders; an arcane and often illogical system of service valuation and payment; and the lack of a standardized, transparent and validated approach to the measurement of 'value.' Recent discussions on reforming the elements of healthcare delivery have increased focus on these systemic shortcomings and conflicts. As a specialty that is clinically wedded to modern and increasingly expensive technology, radiation oncology has often been singled out for scrutiny. A thorough examination and understanding of the various factors and controversies involved in technology development, implementation and valuation analysis is essential to rational growth and development of the specialty. PMID- 21625149 TI - Clinical implementation of intensity-modulated arc therapy. AB - Intensity-modulated arc therapy (IMAT) is a rotational approach to radiation therapy delivered on a conventional linear accelerator using a conventional multileaf collimator. There are 2 key advantages of IMAT. First, the rotational nature of the delivery provides great flexibility in shaping each dose distribution. As a result, IMAT can provide dosimetric advantages relative to fixed-field intensity-modulated radiation therapy (IMRT). The second advantage is the highly efficient nature of the delivery. For centers with an active IMRT program, the clinical implementation of IMAT should be relatively straightforward. For clinical implementation of IMAT, it is important to fully characterize the accuracy of the dose model used, and the performance of the quality assurance equipment. PMID- 21625150 TI - 4D imaging and 4D radiation therapy: a New Era of therapy design and delivery. AB - Recently developed 4D CT imaging technologies have shown that significant organ motion can occur within radiotherapy fields during treatment. Most often a result of respiration, this motion can cause dose delivery errors that are clinically significant when unmanaged, as demonstrated in many recent investigations. Motion during the regular breathing cycling is important, but day-to-day breathing variations, as may be caused by changes in residual tidal volume, can cause systematic shifts in tumor position. These may cause delivery misalignments because the tumor is not in the same average location at each treatment. Approaches to management of this motion may involve motion-inclusive planning, gating or tracking. 4D CT has been instrumental in most of these approaches. Given the state of treatment planning software, it is not possible to preplan whether a specific patient would benefit from one or another of these methods. Daily imaging (or use of a nonimage-based system such as Calypso) is necessary to locate the tumor, and the location must be correlated with measurements from a system that tracks breathing motion during treatment delivery. This is typically done using an independent metric that characterizes the breathing cycle (e.g. the height of the abdomen). Only then can the treatment plan be accurately implemented. There are many methods to manage tumor motion, though most are challenging to implement and remain poorly supported by vendors. When determining which system to use, an important distinction between competing approaches is whether they are amplitude- or phase-based. Some implementations may use different approaches for different parts of the treatment planning and delivery process, potentially introducing errors in the characterization of breathing motion. While many advances have been achieved and are discussed here, the development of solid, stable and robust processes to effectively manage breathing motion remains a foremost and continuing challenge in radiotherapy. PMID- 21625151 TI - Locating and targeting moving tumors with radiation beams. AB - 3D knowledge of the tumor position during abdominal and thoracic radiotherapy is an important component of motion management in radiation therapy. A wide variety of real-time position monitoring systems are available or under development. These are based on a diversity of modalities including radiofrequency, radioisotopes, ultrasound and MRI in addition to the optical, kilovoltage and megavoltage imaging systems available on conventional accelerators. These systems are also providing new insights into the magnitude and complexity of target and normal tissue motion during a course of therapy, and are driving the development of real-time targeting systems. Real-time targeting devices to align the tumor and the radiation beam have built upon technologies of robots, multileaf collimators, and couch-based and gimbaled positioning systems. The integration and widespread dissemination of systems that locate and target moving tumors are ongoing developments in the early 21st century, and future systems are likely to include the functionality of targeting temporally changing tumors and normal tissue physiology as well as anatomy. PMID- 21625152 TI - Technologies of image guidance and the development of advanced linear accelerator systems for radiotherapy. AB - As advanced radiotherapy approaches for targeting the tumor and sparing the normal tissues have been developed, the image guidance of therapy has become essential to directing and confirming treatment accuracy. To approach these goals, image guidance devices now include kV on-board imagers, kV/MV cone-beam CT systems, CT-on-rails, and mobile and in-room radiographic/fluoroscopic systems. Nonionizing sources, such as ultrasound and optical systems, and electromagnetic devices have been introduced to monitor or track the patient and/or tumor positions during treatment. In addition, devices have been designed specifically for monitoring and/or controlling respiratory motion. Optimally, image-guided radiation therapy systems should possess 3 essential elements: (1) 3D imaging of soft tissues and tumors, (2) efficient acquisition and comparison of the 3D images, and (3) an efficacious process for clinically meaningful intervention. Understanding and using these tools effectively is central to current radiotherapy practice. The implementation and integration of these devices continue to carry practical challenges, which emphasize the need for further development of the technologies and their clinical applications. PMID- 21625153 TI - Helical tomotherapy: image-guided and adaptive radiotherapy. AB - Helical tomotherapy is a treatment device that is designed to deliver intensity modulated radiation therapy treatments. Helical tomotherapy systems have been used to treat a wide spectrum of anatomical sites. In addition to its unique delivery technique, the capability to obtain megavoltage-based CT (MVCT) images is highly integrated into the system's image guidance. The introduction of MVCT imaging into clinical practice has prompted a range of technical and clinical investigations. The image quality, image dose and use of MVCT images for dose calculation have been investigated. At the same time, routine clinical use of MVCT imaging has provided a wealth of clinical experience. Both technical and clinical experiences with the MVCT system will be reviewed in this chapter. PMID- 21625154 TI - The CyberKnife in clinical use: current roles, future expectations. AB - The CyberKnife system deploys a linac mounted on an agile robot and directed under image guidance for stereotactic radiotherapy using nonisocentric beam delivery. A design advantage of the CyberKnife system is its method of active image guidance during treatment delivery. Recent developments in the hardware and software of the system have significantly enhanced its functionality: (a) an optimized path traversal process significantly reduces the robot motion time, resulting in reductions of overall treatment times of at least 5-10 min; (b) to optimize the accuracy of dose calculation in CyberKnife planning/delivery, Monte Carlo algorithms have been introduced; (c) the new IRIS collimator reduces the monitor units required, increases treatment speed and improves conformality and homogeneity of treatment plans; (d) XSight lung tracking, an algorithm for fiducial-less lung tracking, has been developed for peripheral, radio-dense lung tumors with diameters >15 mm; and (e) a sequential optimization planning process incorporates a more flexible approach to optimize the multiple, complex treatment planning criteria used today. The clinical efficacy of CyberKnife radiosurgery for brain/head lesions such as metastases, arteriovenous malformations, acoustic neuromas and meningiomas is well established. Since there is no need for skeletal fixation with the CyberKnife, radiosurgery can be applied to targets beyond the brain, and the technology has been extensively used for stereotactic body radiotherapy, treating targets in many anatomic sites. Currently, clinical studies have been completed or are ongoing for common malignancies including tumors involving the spine, lung, pancreas, liver and prostate. PMID- 21625155 TI - Image guidance and the new practice of radiotherapy: what to know and use from a decade of investigation. AB - Over the past decade, fundamental advances in image-guided radiation therapy (IGRT) have been made that are now being implemented in clinical practice. Imaging technologies to direct and confirm beam accuracy at the time of radiotherapy delivery have been intensively researched and developed. More recently, these imaging data have been used to evaluate and even modify the daily dose delivery of intended treatment plans. The rationale for the use of IGRT, to improve tumor control while limiting normal tissue toxicity, is a universal goal in radiotherapy. Avoidance of unexpected under- or overdosing during treatment is the most important benefit of IGRT, and has led to its integration into the use of advanced radiotherapy planning/delivery technologies for many clinical applications. Evidence-based strategies to effectively use IGRT in the clinic are still emerging. The evolving role of IGRT and some proposed strategies to exploit its potential benefits in the clinic will be presented, emphasizing the perspective of the radiation clinician. Practical strategies will be proposed to exploit the potential benefits of IGRT technologies in the clinic. PMID- 21625156 TI - Intensity-modulated and image-guided radiation therapy for head and neck cancers. AB - Radiation therapy is a key component of the multidisciplinary treatment of head and neck cancers (HNC), which are ideal tumors for intensity-modulated radiation therapy (IMRT) because of their location and intimate relationship to the surrounding critical structures. Several institutional studies have suggested that IMRT is superior to conventional radiation therapy in salivary preservation and holds promises for improved locoregional control of these tumors. Small randomized studies have supported the role of IMRT in reducing xerostomia and possibly improving quality of life. Target delineation for IMRT in these tumors is complex and requires detailed knowledge of head and neck anatomy and pathways of tumor spread. The advent of image-guided radiation therapy offers a new innovation that can refine IMRT delivery even further. This article focuses on the issues surrounding IMRT target delineation for typical HNC presentations and a discussion on the role of FDG-PET imaging in HNC treatment planning. PMID- 21625157 TI - Delineating neck targets for intensity-modulated radiation therapy of head and neck cancer. AB - Experience with intensity-modulated radiation therapy (IMRT) for head and neck cancer is building greater understanding of the requirements for therapy planning. Delineation of the lymphatic targets for IMRT of the head and neck is a crucial step in this planning, and often determines the risks of marginal or out of-field local/regional tumor recurrence. Definition of the gross tumor volumes needs to take into account both radiological (CT, MRI, PET) and clinical findings. Understanding of the appropriate CTVs is developing based on: (a) established knowledge of the natural history and spread patterns of head and neck cancer, (b) the accruing experience of clinicians using IMRT, and (c) evaluations of patient outcomes following consistent treatment approaches as determined by institution practice patterns and prospective clinical studies. This chapter will outline the important steps in lymphatic target definition for head and neck cancer, and will discuss several special clinical concerns for these patients and their management. PMID- 21625158 TI - Motion management and image guidance for thoracic tumor radiotherapy: clinical treatment programs. AB - Managing target motion first requires understanding the nature of the motion characteristic of the tumor in the individual patient. It is important to have effective immobilization and patient training strategies to help reduce motion, and then to design appropriate margins and compensation for the residual motion that is quantified. Especially when considering complex, technically demanding treatments that require a degree of patient cooperation, careful patient selection is needed to ensure that the potential benefits of the treatment design are actually realized. Finally, accurate treatment hinges critically on verification - of overall positioning, of target and organ motion at the time of treatment, and of the performance of the selected treatment strategy. Properly selected imaging methods are central to this verification process. This discussion will present practical solutions for motion management and image guidance of radiotherapy for thoracic tumors, and most of these concepts are widely applicable to treatment of other tumor sites as well. PMID- 21625159 TI - Intensity-modulated radiotherapy for breast cancer: advances in whole and partial breast treatment. AB - Intensity-modulated radiotherapy (IMRT) can improve dose distributions through the treated breast and also reduce radiation doses to adjacent normal tissues including the contralateral breast, heart and lung with appropriate planning. Analyses demonstrate that the quality of radiation dose distribution does affect clinical results, and that outcomes are enhanced through improved planning and dose delivery methods. To achieve these results, it is essential to carefully define tissue volumes for treatment or avoidance, select technologies that can potentially conform fields to those volumes, use comprehensive planning methods, and assess their results in terms of objective dose constraints. IMRT can also be used to boost the region of tumor excision concurrently with whole breast treatment, an approach now being evaluated in on-going clinical studies. Partial breast irradiation (PBI) has been proposed as an alternative to irradiation of the entire breast for early-stage breast cancer patients undergoing breast conservation treatment. Numerous single institution phase II studies have demonstrated promising results, and the American Society of Radiation Oncology (ASTRO) has defined a suitable group of low-risk patients for PBI treatment off protocol at this time. IMRT has been proposed as an alternative to 3D conformal radiotherapy (3DCRT) for external beam PBI to improve the dose conformality to target volumes and the sparing of normal tissues. There are an increasing number of institutions evaluating and using IMRT instead of 3DCRT for PBI because of the potential treatment advantages for the breast cancer patient. PMID- 21625160 TI - Image-guided radiotherapy strategies in upper gastrointestinal malignancies. AB - Organ motion due to breathing, peristalsis and deformation presents challenging problems for the delivery of highly conformal radiotherapy to upper abdominal targets, despite the many advancements in the technology of radiation planning and delivery. It is important to understand and account for this motion to avoid treatment inaccuracies, especially systematic errors that could potentially impact the probability of tumor control or increase the risk of normal tissue toxicity. Various image guidance tools can be utilized from the outset of radiation planning through treatment to minimize introducing such errors. These strategies include: assessment of breathing motion (with or without breath hold) prior to simulation, 4D CT simulation and cine MRI to evaluate tumor/organ motion, and image guidance on the treatment unit using kV fluoroscopy and 4D cone beam CT. Together, image guidance methods can provide greater assurance that concordance exists between planned and delivered doses during a course of radiotherapy. PMID- 21625161 TI - Radiotherapy planning for the lymphomas: expanding roles for biologic imaging. AB - Radiotherapy planning now uses advanced technologies to accurately image and assess the extent of disease for treatment. PET scanning has become established as perhaps the most important imaging study for patients with Hodgkin's disease. With respect to initial staging, FDG-PET is more sensitive overall than CT scanning. PET can detect disease at sites that do not meet size criteria by CT. Also, PET is more specific than CT alone because of the functional information that it provides. However, some disease may still escape PET imaging, and false negative results can occur. With respect to treatment response, PET has now become accepted as the most important response measure for the lymphomas. Current protocols are investigating the benefit of this information for radiotherapy planning, and even the possible elimination of radiotherapy in patients completely responding to chemotherapy. For radiotherapy planning, PET/CT should be obtained prior to and after chemotherapy; both scans give important information for the design of the radiation treatment. This chapter will review specific guidelines for planning radiotherapy based on these new imaging capabilities. PMID- 21625162 TI - Image-guided, adaptive radiotherapy of prostate cancer: toward new standards of radiotherapy practice. AB - The development and acceptance of new image-guided radiotherapy (IGRT) technologies have often been initiated with the treatment of prostate cancer. Imaging and tracking of the prostate during a treatment course has yielded a great deal of information about the motion and deformation of the gland during radiotherapy, and has led the way toward the development of more accurate treatment methods including dose-guided and adaptive strategies. Now, there is long-term experience with the use of fiducials and electromagnetic implantable beacons that give high-quality tracking of prostate motion. From analyzing these extensive tracking data sets, a clear understanding of prostate motion and its dosimetric significance has developed. This knowledge can now be used to define current expectations and guidelines for clinical care. The random nature of prostate motion requires daily localization if treatment is to be delivered with small margins. Interfraction motion can have a significant impact on prostate gland dosimetry, and even more of an impact on the seminal vesicles and possibly intraprostatic tumor areas. The dosimetric impact on normal structures (bladder/rectum) is less clear, and there are significant individual variations. Interfraction and intrafraction rotations and deformations of the prostate are routinely detected. The dosimetric impact of these motions of the prostate gland is minimal when daily localization is used, even when the treatment margins are small. However, deformations of the seminal vesicles, rectum and bladder are much more pronounced. The dosimetric impact of deformation of the rectum and bladder is highly variable among patients, and the clinical consequences remain unclear. Daily volumetric imaging and dosimetry may become quite important for these volumes. Due to the random nature of motion/deformation during prostate radiotherapy, adaptive radiotherapy ideally would be performed as an on-line process. On-line adaptive radiotherapy requires robust deformable registration and replanning programs. These are beginning to emerge in useful clinic applications. PMID- 21625163 TI - The expanding roles of stereotactic body radiation therapy and oligofractionation: toward a new practice of radiotherapy. AB - The range of clinical applications for stereotactic body radiation therapy (SBRT) continues to expand based on clinical outcomes data from prospective trials and carefully analyzed institutional experiences. As a result of this strong scientific foundation, there has been burgeoning implementation of SBRT and other forms of hypofractionated radiation therapy in the practice of radiation oncology worldwide. In spite of the clinical successes achieved thus far - or, perhaps, because of them - fundamental questions about SBRT remain and have come into greater focus. Where and when is SBRT optimally integrated into the range of evolving modern multidisciplinary cancer treatment programs? What scientific insights (biological, technical and medical) might lead to further improvements in the efficacy of SBRT? What efficiencies are needed to achieve greater availability of SBRT? These and many other questions, fueled by the clinical accomplishments of SBRT to date, provide compelling directions for further exploration in scientific and clinical studies and further contributes to discoveries already transforming the practice of radiation oncology. PMID- 21625164 TI - Stereotactic body radiation therapy: normal tissue and tumor control effects with large dose per fraction. AB - Stereotactic body radiation therapy (SBRT) is a potent noninvasive means of administering high radiation doses to demarcated tumor deposits in extracranial locations. The treatments use image guidance and related advanced treatment delivery technologies for the purpose of escalating the radiation dose to the tumor, while sharply minimizing the radiation doses to surrounding normal tissues. The local tumor control outcomes for SBRT have been higher than any previously published for the radiotherapy of frequently occurring carcinomas. In addition, the pattern, timing and severity of the toxicities have been very different than from those seen with conventional radiotherapy. These issues pose challenges to our understanding of the radiobiological mechanisms and the optimal uses of SBRT. In this review, the clinical characteristics and outcomes of SBRT are presented in the context of their possible underlying mechanisms. While some of these considerations remain theoretical, they may outline at least qualitative understandings of the observed clinical effects, and motivate continuing research into the effects of SBRT that guide its most effective use in the clinic. PMID- 21625165 TI - Stereotactic body radiation therapy for thoracic cancers: recommendations for patient selection, setup and therapy. AB - Advanced technologies have facilitated the development of stereotactic body radiation therapy (SBRT) programs capable of delivering ablative radiation doses for the control of lung cancers. To date, experience with these programs has been highly favorable, as reflected in the results of careful clinical trials. The medically inoperable lung cancer patient, lacking more effective options, has served as the initial clinical base to test SBRT; the therapeutic outcomes have confirmed a significant role for this approach. For many patient groups, SBRT may become a noninvasive alternative to some thoracic surgeries, especially ones with more limited therapeutic goals such as wedge resection. Despite these results, long-term evaluation of the cases treated is required to allow greater understanding of the limitations and contributions of this new modality. The successful delivery of SBRT requires the development of a comprehensive, specialized clinical program providing advanced technology and the technical expertise of physicians, physicists and therapists specially trained in SBRT applications. To achieve successful clinical outcomes, careful patient selection and attention to therapy design and delivery are required since exacting clinical procedures are involved. This chapter will outline many details essential for establishing an effective SBRT program in clinical practice. PMID- 21625166 TI - Stereotactic body radiation therapy for gastrointestinal malignancies. AB - Stereotactic body radiotherapy (SBRT) is an emerging treatment for pancreas cancer and liver tumors. Early data suggest excellent control rates for locally advanced pancreas cancer. However, due to the close proximity of the duodenum and stomach, steps to effectively minimize toxicities must be taken through image guidance of treatments. SBRT for liver tumors has also shown high rates of local control with low risks for hepatic toxicity. Careful selection of cases for SBRT is essential to achieve disease control and to minimize toxicity for patients. In treatment, attention must be paid to minimizing exposure of nearby normal tissues, including ribs, skin and bowel as well as the functioning organs surrounding the tumors. There is no accepted standard for the SBRT dose/fractionation schedule for these cases and the optimal strategy will likely depend on the size, number and location of lesions for each patient. However, the published data seem to suggest an overall dose-response effect. To realize the clinical potential of SBRT for these tumors, investigations are needed to determine optimum fractionation schedules and to integrate its use with systemic chemotherapy programs. PMID- 21625167 TI - Stereotactic body radiotherapy for prostate cancer: current results of a phase II trial. AB - The hypofractionation of stereotactic body radiotherapy (SBRT) for prostate cancer has become a broad topic, and there are many aspects to consider before accepting this treatment into our clinics. Among the considerations are the data from the Stanford phase II trial, a seminal investigation into this area, which will be presented and reviewed here. A single-arm, prospective phase II trial was initiated at Stanford in December of 2003. This trial uses SBRT as monotherapy for 'low-risk' prostate cancer patients, and 69 patients have been entered to date. We have analyzed the patient data for the first 5 years of this study. For study entry, patients were required to have clinical stage T1c or T2a disease, prostate-specific antigen (PSA) <= 10 and a Gleason score of 3 + 3 (or 3 + 4 if the higher grade portion was of small volume, usually <25% of the cores involved). No prior treatment was permitted, including the use of transurethral resections or androgen deprivation therapies. A low urinary IPSS score of < 20 was required for study entry as well. The prescription dose was 7.25 Gy for 5 fractions for a total dose of 36.25 Gy. This was normalized to cover >= 95% of the planning target volume with 100% of the prescription dose. Patients were treated using CyberKnife technology. To date, excellent PSA responses have been observed in patients with lower-risk disease selected for treatment and receiving 36.25 Gy in 5 fractions. To date, sexual quality of life outcomes have also been approximately comparable to other radiotherapy approaches. Rates of late GI and GU toxicity have been relatively low and generally comparable to dose-escalated approaches using conventional fractionation. PMID- 21625168 TI - Proton therapy: clinical gains through current and future treatment programs. AB - Proton beams can provide a substantial dosimetric advantage because of their unique depth-dose characteristics, which can be exploited to achieve significant reductions in normal tissue doses proximal and distal to the target volume. These may allow escalation of tumor doses, potentially improving local control and survival while at the same time reducing toxicity and improving quality of life. While many of the steps in proton and photon treatment planning processes are similar, there are also significant differences. Some of these arise from the unique physical characteristics of protons, while others are the result of their greater vulnerability to uncertainties, especially from inter- and intrafractional variations in anatomy. These factors must be considered in designing margins and field-shaping devices, as well as in designing treatment plans as a whole and in evaluating them. Ongoing research is aimed at better estimation of these uncertainties and their impact on proton therapy, and reducing these uncertainties through image guidance, adaptive radiotherapy and the development of novel imaging devices and dose computation algorithms. For proton therapy delivery, intensity modulation techniques are already in use, and will continue to be developed and utilized increasingly. The advantages include greater flexibility in dose shaping for improved target coverage and reduced normal tissue dose, potential improvement in plan robustness, and improvement in clinical efficiency. A spectrum of imaging techniques can now be used to assist our understanding of proton dosimetry in the patient, and PET imaging is the one that is furthest developed toward the goal of in vivo dose imaging. To decrease the cost of proton therapy and increase its availability, many technical improvements and practical delivery technologies are being developed, including compact proton machines that will soon become clinically available. PMID- 21625169 TI - Proton therapy in the clinic. AB - The clinical advantage for proton radiotherapy over photon approaches is the marked reduction in integral dose to the patient, due to the absence of exit dose beyond the proton Bragg peak. The integral dose with protons is approximately 60% lower than that with any external beam photon technique. Pediatric patients, because of their developing normal tissues and anticipated length of remaining life, are likely to have the maximum clinical gain with the use of protons. Proton therapy may also allow treatment of some adult tumors to much more effective doses, because of normal tissue sparing distal to the tumor. Currently, the most commonly available proton treatment technology uses 3D conformal approaches based on (a) distal range modulation, (b) passive scattering of the proton beam in its x- and y-axes, and (c) lateral beam-shaping. It is anticipated that magnetic pencil beam scanning will become the dominant mode of proton delivery in the future, which will lower neutron scatter associated with passively scattered beam lines, reduce the need for expensive beam-shaping devices, and allow intensity-modulated proton radiotherapy. Proton treatment plans are more sensitive to variations in tumor size and normal tissue changes over the course of treatment than photon plans, and it is expected that adaptive radiation therapy will be increasingly important for proton therapy as well. While impressive treatment results have been reported with protons, their cost is higher than for photon IMRT. Hence, protons should ideally be employed for anatomic sites and tumors not well treated with photons. While protons appear cost-effective for pediatric tumors, their cost-effectiveness for treatment of some adult tumors, such as prostate cancer, is uncertain. Comparative studies have been proposed or are in progress to more rigorously assess their value for a variety of sites. The utility of proton therapy will be enhanced by technological developments that reduce its cost. Combinations of 3D protons with IMRT photons may offer improved treatment plans at lower cost than pure proton plans. Hypofractionation with proton therapy appears to be safe and cost-effective for many tumor sites, such as for selected liver, lung and pancreas cancers, and may yield significant reduction in the cost of a therapy course. Together, these offer practical strategies for expanding the clinical availability of proton therapy. PMID- 21625171 TI - To give or not to give? Lessons from the arginine paradox. AB - Arginine is one of the 20 amino acids (AA) found in proteins and synthesized by human cells. However, arginine is also the substrate for a series of reactions leading to the synthesis of other AA and is an obligatory substrate for two enzymes with diverging actions, arginases and nitric oxide synthases (NOS), giving origin to urea and NO, respectively. NO is a very potent vasodilator when produced by endothelial NOS (eNOS). The 'arginine paradox' is the fact that, despite intracellular physiological concentration of arginine being several hundred micromoles per liter, far exceeding the ~5 MUM K(M) of eNOS, the acute provision of exogenous arginine still increases NO production. Clinically, an additional paradox is that the largest controlled study on chronic oral arginine supplementation in patients after myocardial infarction had to be interrupted for excess mortality in treated patients. Expression and activity of arginases, which produce urea and divert arginine from NOS, are positively related to exogenous arginine supplementation. Therefore, the more arginine is introduced, the more it is destroyed, eventually leading to impaired NO production. In this review, conditions influencing the low arginine concentrations found in plasma will be reviewed, revising the paradigm that simple replenishment of what is lacking will always produce beneficial consequences. PMID- 21625170 TI - Nutrigenetics and nutrigenomics: viewpoints on the current status and applications in nutrition research and practice. AB - Nutrigenetics and nutrigenomics hold much promise for providing better nutritional advice to the public generally, genetic subgroups and individuals. Because nutrigenetics and nutrigenomics require a deep understanding of nutrition, genetics and biochemistry and ever new 'omic' technologies, it is often difficult, even for educated professionals, to appreciate their relevance to the practice of preventive approaches for optimising health, delaying onset of disease and diminishing its severity. This review discusses (i) the basic concepts, technical terms and technology involved in nutrigenetics and nutrigenomics; (ii) how this emerging knowledge can be applied to optimise health, prevent and treat diseases; (iii) how to read, understand and interpret nutrigenetic and nutrigenomic research results, and (iv) how this knowledge may potentially transform nutrition and dietetic practice, and the implications of such a transformation. This is in effect an up-to-date overview of the various aspects of nutrigenetics and nutrigenomics relevant to health practitioners who are seeking a better understanding of this new frontier in nutrition research and its potential application to dietetic practice. PMID- 21625172 TI - The basis of differential responses to folic acid supplementation. AB - BACKGROUND/AIMS: Elevated levels of total homocysteine (tHcy) are associated with an increased risk of many common diseases. Supplementation with folic acid has been shown to significantly reduce tHcy levels. We used the classical twin model to partition the variability in changes in plasma tHcy levels through folic acid supplementation into genetic, environmental, and confounding epidemiological factors. METHODS: We carried out an intervention study of folic acid using 101 healthy, female, identical and non-identical twins aged 50-80 years. Each twin was administered folic acid (0.8 mg/day) for 6 weeks. Total plasma folate, cobalamin and tHcy were measured at both baseline and after dosing. We calculated the heritability and tested for associations between the MTHFR C677T functional variant and response to folic acid supplementation. RESULTS: Supplementation with folic acid led to a significant reduction in tHcy levels. The mean tHcy changed from 12.14 to 10.42 MUmol/l after supplementation (p < 10(-5)). Moreover, the change in tHcy levels was highly heritable (64%), not associated with the C677T functional variant at MTHFR and not confounded by age, BMI or diet. CONCLUSIONS: Our results highlight the need to identify genetic factors associated with biomarkers of response to folate supplementation. PMID- 21625173 TI - Determinants of aspirin metabolism in healthy men and women: effects of dietary inducers of UDP-glucuronosyltransferases. AB - BACKGROUND/AIMS: Interindividual variation in aspirin (ASA) metabolism is attributed to concomitant use of drugs or alcohol, urine pH, ethnicity, sex, and genetic variants in UDP-glucuronosyltransferases (UGT). Little is known about the effects of diet. METHODS: We evaluated cross-sectionally whether urinary excretion of ASA and its metabolites [salicylic acid (SA), salicyluric acid (SUA) phenolic glucuronide (SUAPG), salicylic acid acyl glucuronide (SAAG) and salicylic acid phenolic glucuronide (SAPG)] differed by UGT1A6 genotype and dietary factors shown to induce UGT. Following oral treatment with 650 mg ASA, urine was collected over 8 h in 264 men and 264 women (21-45 years old). RESULTS: There were statistically significant differences in metabolites excreted between sexes and ethnicities. Men excreted more SUA; women more ASA (p = 0.03), SA, SAAG and SAPG (p <= 0.001 for all). Compared to Caucasians, Asians excreted more ASA, SA and SAAG, and less SUA and SUAPG (p <= 0.03 for all); African-Americans excreted more SAAG and SAPG and less SUA (p <= 0.04). There was no effect of UGT1A6 genotypes. Increased ASA and decreased SUAPG excretion was observed with increased servings of vegetables (p = 0.008), specifically crucifers (p = 0.05). CONCLUSION: Diet may influence the pharmacokinetics of ASA, but effects may be through modulation of glycine conjugation rather than glucuronidation. PMID- 21625174 TI - Aging is associated with a proapoptotic endothelial progenitor cell phenotype. AB - The aim of this study was to determine if aging is associated with enhanced endothelial progenitor cell (EPC) sensitivity to apoptosis. Cells with phenotypic EPC characteristics were isolated from healthy, nonobese young (age 25 +/- 1 years) and older (61 +/- 1 years) men. Intracellular active caspase-3 concentrations in response to staurosporine stimulation were approximately 35% higher (p < 0.05) in EPCs from older (3.15 +/- 0.29 pg/ml) compared with young (2.33 +/- 0.24 pg/ml) men. Protein expression of Akt, p70 S6-kinase and Bcl-2 was markedly lower (approx. 35, 75 and 60%, respectively, all p < 0.05) in EPCs from older compared with young men, whereas there were no age-related differences in either 14-3-3epsilon or Bax expression. Additionally, EPC telomerase activity was 57% lower (p < 0.05) in older (0.18 +/- 0.11 AU) versus young (0.43 +/- 0.11 AU) men. These results indicate that aging is associated with a proapoptotic EPC phenotype characterized by decreased expression of key antiapoptotic proteins associated with the PI-3-kinase signaling pathway and reduced telomerase activity. These age-related changes likely contribute, in part, to the diminished ability of EPCs to resist an apoptotic stimulus in older men. Increased susceptibility to apoptosis may contribute to the numerical and functional impairments observed in EPCs with aging. PMID- 21625175 TI - Functional characterization of embryonic stem cell-derived endothelial cells. AB - Endothelial cells (EC) derived from embryonic stem cells (ESC) require additional functional characterization before they are used as a cell therapy in order to enhance their potential for engraftment and proliferation. We explore several physiologically relevant functions of ESC-derived EC (ESC-EC), such as its capacity to produce nitric oxide (NO), regulate permeability, activate and express surface molecules for the recruitment of leukocytes in response to inflammatory stimuli, migrate and grow new blood vessels, lay down extracellular matrix, and take up low-density lipoproteins. We also examined the ESC-EC ability to upregulate NO in response to shear stress and downregulate NO in response to pro-inflammatory TNF-alpha activation. Functional responses of ESC-EC were compared with those of cultured mouse aortic ECs. The ESC-EC exhibit most aspects of functional endothelium, but interesting differences remain. The ESC-EC produced less NO on a per cell basis, but the same amount of NO if quantified based on the area of endothelial tissue. They also exhibit increased angiogenic sprouting and are more resistant to inflammatory signals. We further characterized the subphenotype of our ESC-EC and observed both venous and arterial markers on individual cells with a larger percentage of the cells exhibiting a venous phenotype. These data support the hypothesis that the developmental default pathway is toward a venous EC, and that refinement of methods for differentiation towards arterial EC is required to maintain a homogeneous population. PMID- 21625176 TI - Cellular and molecular responses of the basilar terminus to hemodynamics during intracranial aneurysm initiation in a rabbit model. AB - BACKGROUND/AIMS: Hemodynamics constitute a critical factor in the formation of intracranial aneurysms. However, little is known about how intracranial arteries respond to hemodynamic insult and how that response contributes to aneurysm formation. We examined early cellular responses at rabbit basilar termini exposed to hemodynamic insult that initiates aneurysmal remodeling. METHODS: Flow in the basilar artery was increased by bilateral carotid artery ligation. After 2 and 5 days, basilar terminus tissue was examined by immunohistochemistry and quantitative PCR. RESULTS: Within 2 days of flow increase, internal elastic lamina (IEL) was lost in the periapical region of the bifurcation, which experienced high wall shear stress and positive wall shear stress gradient. Overlying endothelium was still largely present in this region. IEL loss was associated with localized apoptosis and elevated expression of matrix metalloproteinases (MMPs) 2 and 9. A small number of inflammatory cells were sporadically scattered in the bifurcation adventitia and were not concentrated in regions of IEL loss and MMP elevation. Elevated MMP expression colocalized with smooth muscle alpha-actin in the media. CONCLUSION: The initial vascular response to aneurysm-initiating hemodynamic insult includes localized matrix degradation and cell apoptosis. Such destructive remodeling arises from intrinsic mural cells, rather than through inflammatory cell infiltration. PMID- 21625177 TI - Characterization of human late outgrowth endothelial progenitor-derived cells under various flow conditions. AB - BACKGROUND: Endothelial progenitor-derived cells (EPC) are a cell therapy tool in peripheral arterial disease and for re-endothelialization of bypasses and stents. OBJECTIVE: To assess EPC behavior under flow conditions normally found in vivo. RESULTS: EPC were isolated from human cord blood, cultured on compliant tubes and exposed in an in vitro flow system mimicking hemodynamic environments normally found in medium and large arteries. EPC exposed for 24 h to unidirectional (0.3 +/- 0.1 or 6 +/- 3 dynes/cm(2)) shear stress oriented along flow direction, while those exposed to bidirectional shear stress (0.3 +/- 3 dynes/cm(2)) or static conditions had random orientation. Under bidirectional flow, tissue factor (TF) activity and mRNA expression were significantly increased (2.5- and 7.0-fold) compared to static conditions. Under low shear unidirectional flow TF mRNA increased 4.9 +/- 0.5-fold. Similar flow-induced increases were observed for TF in mature umbilical vein-derived endothelial cells. Expression of tissue-type plasminogen activator (t-PA), urokinase (u-PA) and monocyte chemotactic protein 1 (MCP1) were reduced by 40-60% in late outgrowth endothelial progenitor-derived cells (LO-EPC) exposed to any flow environment, while MCP1, but not t-PA or u-PA, was decreased in HUVEC. CONCLUSIONS: Flow, in particular bidirectional, modifies the hemostatic balance in LO-EPC with increased TF and decreased plasminogen activator expression. PMID- 21625178 TI - A mother and daughter with unexplained renal failure. PMID- 21625179 TI - A phase I study of the safety and pharmacokinetics of the hypoxia-activated prodrug TH-302 in combination with doxorubicin in patients with advanced soft tissue sarcoma. AB - PURPOSE: The purpose of this study was to determine the dose-limiting toxicities (DLT), maximum tolerated dose (MTD), safety, pharmacokinetics and preliminary activity of TH-302, a hypoxia-activated prodrug, in combination with doxorubicin in patients with advanced soft tissue sarcoma. PATIENTS AND METHODS: TH-302 was administered intravenously on days 1 and 8 and doxorubicin 75 mg/m2 on day 1 (2 h after TH-302) of every 3-week cycle. TH-302 starting dose was 240 mg/m2 with a classic 3 + 3 dose escalation. Pharmacokinetics were assessed on days 1 and 8 of cycle 1. Tumor assessments were performed after every second cycle. RESULTS: Sixteen patients enrolled. Prophylactic growth factor support was added due to grade 4 neutropenia. The MTD was 300 mg/m2. DLTs at 340 mg/m2 were neutropenia associated infection and grade 4 thrombocytopenia. Common adverse events included fatigue, nausea and skin rash. There was no evidence of pharmacokinetic interaction between TH-302 and doxorubicin. Five of 15 (33%) evaluable patients had a partial response by RECIST (Response Evaluation Criteria in Solid Tumors) criteria. CONCLUSIONS: The hematologic toxicity of doxorubicin is increased when combined with TH-302. This can be mitigated by prophylactic growth factor support. Toxicities were manageable and there was evidence of antitumor activity. PMID- 21625180 TI - Prognostic relevance of circulating tumor cells in metastatic uveal melanoma. AB - OBJECTIVE: Uveal melanoma primarily metastasizes hematogenously with metastases often confined to the liver. The aim of this study was to investigate the presence of circulating tumor cells (CTC) in patients with metastatic disease as a marker for systemic disease and to determine their prognostic relevance. METHODS: Blood samples from 68 patients were collected at the time of initial treatment of metastases. mRNA expression of tyrosinase and MelanA/MART1 as a surrogate marker for the presence of CTC was analyzed by real-time RT-PCR and compared with patient characteristics. RESULTS: CTC were detected in 63% of all patients and in 67% of the 48 patients with only liver metastases. Univariate and multivariate analyses revealed PCR results and serum lactate dehydrogenase as independent prognostic factors for progression-free (hazard ratios 2.2/3.5) and overall survival (hazard ratios 4.0/6.5). Combination of PCR and lactate dehydrogenase divided the patient cohort into 3 groups with distinct prognosis. CONCLUSION: CTC as evidence for systemic disease can be found in the majority of patients with metastatic uveal melanoma, including patients with visible disease confined to the liver. Detection of CTC-specific mRNA transcripts for tyrosinase and MelanA/MART1 by PCR is a poor prognostic factor for progression-free and overall survival. Characterization of CTC could improve the understanding of their biology. PMID- 21625181 TI - Efficacy of sunitinib rechallenge in kidney cancer: are mTOR inhibitors involved or is it only a matter of time? PMID- 21625182 TI - Predictive value of F-18 FDG PET/CT for malignant pleural effusion in non-small cell lung cancer patients. AB - BACKGROUND: The purpose of this study was to evaluate the accuracy of F-18 fluorodeoxyglucose positron emission tomography/computed tomography (F-18 FDG PET/CT) imaging in the detection of malignant pleural effusion and pleural metastasis in patients with non-small cell lung cancer (NSCLC). MATERIALS AND METHODS: We analyzed F-18 FDG PET/CT images of 33 lung cancer patients with pleural effusion. We used 2 categorical parameters to differentiate malignant from benign pleural effusion: i) quantitative parameters using maximum standardized uptake value (SUVmax of effusion and pleura, and the following ratios: lesion to aorta (L/Ao), to cerebellum (L/Cbl), to liver (L/Liv), to nonlesion (L/NL), and to primary lung cancer (L/Prim)) and ii) various parameters determined by PET and CT scans (uptake at the pleural region, Hounsfield unit, size, and morphology of any solid abnormality). RESULTS: Malignant pleural effusions showed significantly higher L/Prim values than benign pleural effusions. The presence of pleural abnormality on CT and pleural region uptake on PET images were found to be significantly more frequent in cases of malignant pleural disease. These parameters could differentiate malignant and benign pleural effusion according to receiver operating characteristic (ROC) analyses. There were no statistical differences between L/Prim, pleural abnormality on CT, and pleural region uptake on PET images. Abnormal pleural region uptake on PET images was the most accurate parameter identifying malignant pleural effusion by logistic regression analysis. CONCLUSIONS: Our results suggest that F-18 FDG PET/CT can be used as a reliable and noninvasive method for the differentiation of malignant and benign pleural disease in patients with NSCLC. PMID- 21625183 TI - Management of patients with brain metastases receiving trastuzumab treatment for metastatic breast cancer. AB - BACKGROUND: With the more effective control of visceral metastases in patients with metastatic breast cancer (MBC), an increasing number of patients face brain metastases (BM). The aim of this retrospective analysis was to investigate the incidence and factors affecting the prognosis of patients with BM under trastuzumab treatment for MBC. PATIENTS AND METHODS: A total of 75 HER2positive patients treated with trastuzumab for MBC were included. RESULTS are discussed in the context of the current literature. RESULTS: Patients who developed BM (n = 29) had longer median progression-free survival (PFS) during first-line chemotherapy and longer overall survival (OS) after diagnosis of MBC than 46 patients without BM (PFS: 27 vs. 14 months, p = 0.039; OS: 46 vs. 18 months, p = 0.067). Median survival of patients with continuation of trastuzumab after diagnosis of BM was longer than survival of patients with discontinuation of trastuzumab treatment after BM (18 vs. 3 months, p = 0.006). Survival of patients who were treated with surgery and radiotherapy for BM was better compared with radiotherapy alone (9 vs. 5 months, p = not significant) or best supportive care (9 vs. 2 months, p = 0.049). CONCLUSIONS: Continuation of trastuzumab treatment as well as resection of BM seem to give further benefit in the treatment of patients with HER2-overexpressing MBC. PMID- 21625184 TI - Efficacy of sunitinib re-exposure after failure of an mTOR inhibitor in patients with metastatic RCC. AB - BACKGROUND: The sequential use of tyrosine kinase inhibitors (TKI) followed by mTOR inhibitors (mTORi) has been recently established for the systemic treatment of metastatic renal cell carcinoma (mRCC). However, subsequent treatment in mTORi refractory disease remains undetermined. We analyzed the efficacy of sunitinib re challenge after failure of an mTORi at 2 German centers. PATIENTS AND METHODS: Thirteen patients who failed both sunitinib and an mTORi were analyzed, and all patients were re-exposed to sunitinib. Tumor assessment was performed every 2nd cycle of sunitinib or every 3 months. Tumor response was assessed according to RECIST criteria. RESULTS: Initial treatment with sunitinib was associated with a median progression free survival (PFS) of 21 months. Objective response consisted of 2 (15%) complete remissions and 7 (54%) partial remissions (PR) as best response. At the time of re-exposure, 12 of 13 (92%) patients again showed clinical benefit which was associated with a median PFS of 6.9 months and consisted of 2 (15%) PR and 10 (77%) disease stabilizations. CONCLUSIONS: In sunitinib-responsive patients, re-challenge with sunitinib has been successfully introduced after mTORi-refractory disease, underscoring the at least partially transient nature of TKI resistance in mRCC. PMID- 21625185 TI - Very late and isolated leptomeningeal relapse of a pulmonary adenocarcinoma presenting as dementia. PMID- 21625186 TI - Rapid-onset, prolonged bone marrow failure following rituximab therapy of follicular lymphoma. PMID- 21625187 TI - Long-term survival in a smoking caucasian male patient treated with gefitinib for spinal cord compression secondary to lung cancer. PMID- 21625188 TI - Lymph node collision tumor of non-small cell lung carcinoma and chronic lymphocytic leukemia. PMID- 21625189 TI - Potential pitfalls in the management of primary pancreatic cystic neoplasms. AB - Primary pancreatic cystic neoplasms (PCN) have been increasingly recognized over the last 2 decades, mainly as a result of the widespread use of modern imaging techniques in patients with abdominal complaints. Biological behavior and management greatly differ between subgroups of PCN. Accurate preoperative diagnosis is required to select the optimal management strategy for each individual patient with PCN. An in-depth knowledge of the biological, imaging, macroscopic/microscopic features, and laboratory findings of each subtype of PCN is required on the part of the clinician, in order to select a cost-effective diagnostic evaluation of the patient. Clinical judgment is also required on the part of the surgeon to select the optimal procedure and to avoid severe pitfalls in the surgical management of these neoplasms. PMID- 21625190 TI - 'Doc, is palliative care an option for me?'. PMID- 21625191 TI - Impact of intravitreal injection of tissue plasminogen activator on full-field electroretinogram in patients with macular oedema secondary to retinal vein occlusion. AB - PURPOSE: To evaluate the retinal toxicity of intravitreal tissue plasminogen activator (tPA) injection for branch and central retinal vein occlusion (BRVO and CRVO) using the electroretinogram (ERG). PROCEDURES: Ten BRVO patients and 5 CRVO patients were enrolled. A complete examination including full-field ERG, visual acuity, central retinal thickness (CRT), and evaluation of systemic and ocular complications was performed before and after intravitreal tPA injection. RESULTS: No significant differences were found in the amplitude or implicit time of any ERG component after tPA injection, and no systemic or ocular complication was observed. The improvement of visual acuity was significant at month 3 in the BRVO group (p < 0.05) but not in the CRVO group. CRT significantly decreased over the course of 3 months in both groups (p < 0.01). CONCLUSION: Intravitreal tPA injection seems to be a safe and effective treatment option for macular oedema caused by retinal vein occlusions. PMID- 21625192 TI - Oral squamous cell carcinoma: clinicopathological features in patients with and without recurrence. AB - AIM: To compare the clinicopathological profile of oral squamous cell carcinoma (OSCC) in groups with and without recurrence. METHODS: Records of all patients who underwent surgery for primary OSCC at a single institution during 1999 were identified. Patient demographics, lesion site, clinical and pathologic stage, pathologic grading, pattern of invasion, lymphocytic infiltrate, perineural invasion, and treatment and survival data were collected. Descriptive statistics were calculated for each variable and survival was calculated using Kaplan-Meier and Cox models. Patients were divided into 2 groups: with (n = 25) and without (n = 28) recurrence. RESULTS: Tongue (p = 0.02) and poorly differentiated (p = 0.04) tumors were associated with recurrence. Kaplan-Meier and Cox models revealed tobacco use and the absence of lymphocytic infiltrate to be associated with the poorest survival in recurrent OSCC. CONCLUSION: The tumor site, tobacco use, and pathological features were involved in the recurrence of OSCC and should be taken into account for OSCC treatment and follow-up. PMID- 21625193 TI - Comparison of ultrasound-guided biopsy technique for thyroid nodules with respect to adequacy of cytological material. AB - BACKGROUND: Ultrasound-guided fine-needle aspiration cytology (FNAC) sampling of the thyroid represents a standard diagnostic procedure in the evaluation of thyroid nodules. The specimen can be acquired using either of two different techniques. In the first, the short axis is used with observation only of the tip of the needle whilst in the nodule. In the second technique, the long axis is used with the observation of the entire length of the needle. The decision to sample utilizing either technique was done randomly. This study is a retrospective review performed to compare these two techniques with regard to specimen adequacy. METHODS: Ultrasound-guided FNACs were performed in 80 thyroid nodules between May 2008 and February 2009. One physician acquired the cytology specimens using one of these two methods after localization. Data on the type of technique and its diagnostic accuracy were collected. RESULTS: Forty-nine of 80 thyroid nodules were sampled using the long-axis technique. The overall and deep lesion diagnostic adequacies of specimens were significantly higher using this technique (93.9 and 95.1%, respectively, p < 0.01) than the short-axis technique. When comparing the long and short axes for superficial lesions, there was no significant difference in adequacy of the samples (p = 0.92). CONCLUSIONS: This is the first study to compare long- and short-axis techniques with regard to specimen adequacy for thyroid nodules. The long-axis technique decreased the rate of inadequate material and provided more accurate cytological evaluation for deeper lesions. PMID- 21625194 TI - Correlation of routine examinations for the diagnosis of house dust mite allergic rhinitis. AB - OBJECTIVE: To evaluate the correlation between the routine diagnostic tests for house dust mite (HDM) sensitization in Chinese allergic rhinitis (AR) patients, including the skin prick test (SPT), serum HDM-specific immunoglobulin E (sIgE) and nasal smear eosinophilia. METHODS: The ImmunoCAP assay was employed to detect serum total IgE and sIgE. The SPT and nasal smear eosinophil counts were performed using standard procedures. RESULTS: Of 82 subjects with HDM-related symptoms and positive SPT results, 55 individuals showed an sIgE level equal to or greater than CAP class 1 for both Dermatophagoides pteronyssinus (Der p) and D. farinae (Der f), and 62 individuals demonstrated positive eosinophils in nasal smears. sIgE of Der p and Der f presented a linear correlation, and the level of serum HDM-sIgE was also positively correlated with total IgE. An increasing sIgE level was correlated with higher SPT grades in both Der p and Der f. Meanwhile, the correlation coefficient exhibited an increase in value with aging, and the 31 to 40-year age group demonstrated the highest value for both Der p and Der f. CONCLUSION: The positive correlation between SPT and sIgE reaction to HDM varied according to the age of patients with AR. PMID- 21625195 TI - Morphologic patterns of autoimmune pancreatitis in CT and MRI. AB - BACKGROUND/AIMS: To retrospectively evaluate the morphologic characteristics of autoimmune pancreatitis (AIP) using MRI and CT. METHODS: 86 dynamic contrast enhanced CT and MRI scans in 36 AIP patients were evaluated regarding: different enlargement types, abnormalities of the main pancreatic duct (MPD), morphology of the parenchyma and other associated findings. RESULTS: 3 types of enlargement were found: (1) a focal type (28%), (2) a diffuse type (involving the entire pancreas, 11%) and (3) a combined type (56%). The MPD was usually dilated together with focal or diffuse narrowing in 67% (24/36). Unenhanced MRI showed AIP area in 56% (mostly T(1) hypo- and T(2) hyperattenuating), and CT in 10% (hypoattenuating). The arterial phase depicted similar patterns for CT and MRI (hypoattenuating in 58 and 52%, respectively). Venous and late venous phase patterns were usually hyperattenuating in MRI (65 and 74%, late enhancement), while CT mostly showed no signal differences (isoattenuating in 57 and 75%), yielding significant differences between CT and MRI for the venous (p < 0.0001) and the late phase (p = 0.025). Miscellaneous findings were: rim sign (25%), pseudocysts (8%) and infiltration of large vessels (11%). CONCLUSIONS: The 'late enhancement' sign seems to be a key feature and is best detectable with MRI. MRI may be recommended in the diagnostic workup of AIP patients. and IAP. PMID- 21625196 TI - The role of phosphatidylinositol 3-kinase signaling pathways in pancreatic cancer. AB - BACKGROUND: Pancreatic cancer is a highly malignant cancer and the fourth leading cause of cancer-related death. It is characterized by a rapid disease progression, a highly invasive tumor phenotype, and frequently resistance to chemotherapy. Despite significant advances in diagnosis, staging, and surgical management of the disease during the past decade, prognosis of pancreatic cancer is still dismal. METHODS AND RESULTS: The phosphatidylinositol 3-kinase (PI3K) signaling pathways regulate cellular growth, metabolism, survival, and motility in pancreatic cancer. Pancreatic cancer is associated with a high degree of genetic alterations that can result in aberrant activation of the PI3K signaling pathway. Elucidating the role of the PI3K signaling pathway in pancreatic cancer may thus be both meaningful and necessary. CONCLUSION: Improved knowledge of the PI3K signaling pathway in pancreatic cancer would furthermore be helpful in understanding mechanisms of tumor initiation and progression, and in identifying appropriate targeted anticancer treatment in pancreatic cancer. and IAP. PMID- 21625197 TI - Autoimmune pancreatitis in Hungary: a multicenter nationwide study. AB - BACKGROUND: To date, most cases of autoimmune pancreatitis (AIP) have been reported from Japan. The aim of the present study was to assess the clinical features and management of AIP cases in Hungary. METHODS: The demographics, clinical presentation, laboratory and imaging findings, extrapancreatic involvement, treatment response and recurrence were evaluated in the first 17 patients diagnosed with AIP in Hungary. RESULTS: The mean age at presentation was 42.7 years (range: 16-74); 47% of the patients were women. New-onset mild abdominal pain (76%), weight loss (41%) and jaundice (41%) were the most common symptoms, with inflammatory bowel disease being the most frequent (36%) extrapancreatic manifestation. Diffuse pancreatic swelling was seen in 7 patients (41%) and a focal pancreatic mass in 8 (47%). Endoscopic retrograde cholangiopancreatography revealed pancreatic duct strictures in all study patients. The serum IgG4 level at presentation was elevated in 62% of the 8 patients in whom it was measured. All the percutaneous core biopsies (5 patients) and surgical specimens (2 patients), and 2 of the 4 biopsies of the papilla of Vater revealed the typical characteristic findings of AIP: a diffuse lymphoplasmacytic infiltration, marked interstitial fibrosis and obliterative phlebitis. Immunostaining indicated IgG4-positive plasma cells in 62% of the 8 patients in whom it was performed. Granulocytic epithelial lesions (GEL) were present in 3 patients. The patients without GELs were older (mean age 59 years), while those with GEL were younger (mean age 34 years), and 2 of 3 were female and had ulcerative colitis. A complete response to steroid treatment was achieved in all 15 patients. Because of the suspicion of a pancreatic tumor, 2 patients with focal AIP underwent partial pancreatectomy. One patient relapsed, but responded to azathioprine. CONCLUSIONS: This first Hungarian series has confirmed several previously reported findings on AIP. AIP with GEL was relatively frequent among our patients: these patients tended to be younger than in earlier studies and displayed a female preponderance with a high coincidence of ulcerative colitis. Performance of a percutaneous biopsy is strongly recommended. The response to immunosuppressive therapy was excellent. and IAP. PMID- 21625198 TI - Vasonatrin peptide, a new regulator of adiponectin and interleukin-6 production in adipocytes. AB - BACKGROUND: In addition to lipolytic function, ANP plays regulatory roles in the production of various adipokines including adiponectin, leptin, and interleukins. However, the adipose effects of vasonatrin peptide (VNP), a new manmade natriuretic peptide, are largely unknown. AIM: The aim of the present study was to identify the roles of VNP on adipokines production, as well as signaling pathways involved. MATERIAL, SUBJECTS, AND METHODS: 3T3-L1 cells were differentiated into adipocytes and exposed to various concentrations of VNP. Quantitative PCR and immunoassays were performed to determine the mRNA and protein levels of adiponectin and interleukin-6 (IL-6), respectively. The involved signaling pathway was identified by radioimmunoassay to detect the levels of intracellular cyclic GMP (cGMP), mimicking experiments using 8-brcGMP (a membrane-permeable cGMP analog). Also, blocking experiments were performed using HS-142-1, an antagonist of particulate guanylyl cyclase-coupled natriuretic peptide receptor (NPR), or KT-5823, the cGMP-dependent protein kinase (PKG) inhibitor. RESULTS: VNP markedly enhanced adiponectin mRNA expression, as well as protein secretion, however, suppressed IL-6 production in mature adipocytes. In addition, VNP significantly increased the intracellular levels of cGMP. The effects of VNP were mimicked by 8-br-cGMP, whereas inhibited by HS-142-1, or KT 5823. CONCLUSIONS: Taken together, VNP regulates adiponectin and IL-6 production in adipocytes via guanylyl cyclase-coupled NPR/cGMP/PKG pathway. PMID- 21625200 TI - Transplantation: Low serum albumin associated with worse transplant outcome. PMID- 21625199 TI - Chronic kidney disease: Use of urine albumin and cystatin C levels improves risk stratification in chronic kidney disease. PMID- 21625202 TI - Pediatrics: Sirolimus may stunt growth of kidney-transplanted children. PMID- 21625204 TI - Risk factors: Lowering homocysteine levels may have no 'FAVORITable' effect on cardiovascular outcomes. PMID- 21625205 TI - Acute kidney injury: Fluid therapy in acute kidney injury: the FACTTs. PMID- 21625206 TI - Chronic kidney disease: Staging CKD: can we rely on estimated GFRs? PMID- 21625207 TI - Dialysis: Peritoneal dialysis vs hemodialysis: time to end the debate? PMID- 21625208 TI - Dialysis: Two for the price of one: is rt-PA ready for prime time? PMID- 21625209 TI - Role of eIF3a in regulating cisplatin sensitivity and in translational control of nucleotide excision repair of nasopharyngeal carcinoma. AB - Translational control at the initiation step has been recognized as a major and important regulatory mechanism of gene expression. Eukaryotic initiation factor 3a (eIF3a), a putative subunit of the eIF3 complex, has recently been shown to have an important role in regulating the translation of a subset of mRNAs and is found to correlate with the prognosis of cancers. In this study, using nasopharyngeal carcinoma (NPC) cells as a model system, we tested the hypothesis that eIF3a negatively regulates the synthesis of nucleotide excision repair (NER) proteins, and, in turn, cellular response to treatments with DNA-damaging agents such as cisplatin (cis-dichlorodiammine platinum(II) (CDDP)). We found that a CDDP-sensitive sub-clone S16 isolated through limited dilution from an NPC cell line CNE-2 has increased eIF3a expression. Knocking down its expression in S16 cells increased cellular resistance to CDDP, NER activity and synthesis of the NER proteins XPA, XPC, RAD23B and RPA32. Altering eIF3a expression also changed the cellular response to CDDP and UV treatment in other NPC cell lines. Taken together, we conclude that eIF3a has an important role in the CDDP response and in NER activity of NPCs by suppressing the synthesis of NER proteins. PMID- 21625210 TI - LIN28B fosters colon cancer migration, invasion and transformation through let-7 dependent and -independent mechanisms. AB - Lin28b is an RNA-binding protein that inhibits biogenesis of let-7 microRNAs. LIN28B is overexpressed in diverse cancers, yet a specific role in the molecular pathogenesis of colon cancer has to be elucidated. We have determined that human colon tumors exhibit decreased levels of mature let-7 isoforms and increased expression of LIN28B. To determine LIN28B's mechanistic role in colon cancer, we expressed LIN28B in immortalized colonic epithelial cells and human colon cancer cell lines. We found that LIN28B promotes cell migration, invasion and transforms immortalized colonic epithelial cells. In addition, constitutive LIN28B expression increases expression of intestinal stem cell markers LGR5 and PROM1 in the presence of let-7 restoration. This may occur as a result of Lin28b protein binding LGR5 and PROM1 mRNA, suggesting that a subset of LIN28B functions is independent of its ability to repress let-7. Our findings establish a new role for LIN28B in human colon cancer pathogenesis, and suggest LIN28B post transcriptionally regulates LGR5 and PROM1 through a let-7-independent mechanism. PMID- 21625211 TI - COP9 signalosome subunit 6 stabilizes COP1, which functions as an E3 ubiquitin ligase for 14-3-3sigma. AB - 14-3-3sigma, a gene upregulated by p53 in response to DNA damage, exists as part of a positive-feedback loop, which activates p53 and is a human cancer epithelial marker downregulated in various cancer types. 14-3-3sigma levels are critical for maintaining p53 activity in response to DNA damage and regulating signal mediators such as Akt. In this study, we identify mammalian constitutive photomorphogenic 1 (COP1) as a novel E3 ubiquitin ligase for targeting 14-3 3sigma through proteasomal degradation. We show for the first time that COP9 signalosome subunit 6 (CSN6) associates with COP1 and is involved in 14-3-3sigma ubiquitin-mediated degradation. Mechanistic studies show that CSN6 expression leads to stabilization of COP1 through reducing COP1 self-ubiquitination and decelerating COP1's turnover rate. We also show that CSN6-mediated 14-3-3sigma ubiquitination is compromised when COP1 is knocked down. Thus, CSN6 mediates 14-3 3sigma ubiquitination through enhancing COP1 stability. Subsequently, we show that CSN6 causes 14-3-3sigma downregulation, thereby activating Akt and promoting cell survival. Also, CSN6 overexpression leads to increased cell growth, transformation and promotes tumorigenicity. Significantly, 14-3-3sigma expression can correct the abnormalities mediated by CSN6 expression. These data suggest that the CSN6-COP1 axis is involved in 14-3-3sigma degradation, and that deregulation of this axis will promote cell growth and tumorigenicity. PMID- 21625212 TI - Evidence for the transmission of neoplastic properties from transformed to normal human stem cells. AB - The in vivo relationship between human tumor cells and interacting normal cells in their local environment is poorly understood. Here, using a uniquely developed in vitro co-culture system for human embryonic stem cells (hESCs), we examined the interactions between transformed and normal human stem cells. Co-culture of transformed-hESCs (t-hESCs) with normal hESCs led to enhanced self-renewal and niche independence in normal hESCs. Global gene expression analysis of normal hESCs after timed exposure to t-hESCs indicated a transition of the molecular network controlling the hESC state, which included epigenetic changes, towards neoplastic features. These included enhanced pluripotent marker expression and a differentiation blockade as major hallmark changes. Functional studies revealed a loss in normal terminal differentiation programs for both hematopoiesis and neural lineages after normal stem cell co-culture with transformed variants. This transmission of neoplastic properties from t-hESCs to normal hESCs was dependent on direct cell-cell contact. Our study indicates that normal human stem cells can co-opt neoplastic cancer stem cell properties, raising the possibility that assimilation of healthy cells towards neoplastic behavior maybe a contributing feature of sustained tumorigenesis in vivo. PMID- 21625213 TI - HPV16 E5 affects the KGFR/FGFR2b-mediated epithelial growth through alteration of the receptor expression, signaling and endocytic traffic. AB - The E5 oncoprotein of the human papillomavirus type 16 (HPV16 E5) cooperates in cervical carcinogenesis and in epithelial transformation deregulating cell growth, survival and differentiation through the modulation of growth factor receptors. Among the epithelial receptor tyrosine kinases, the keratinocyte growth factor receptor/fibroblast growth factor receptor 2b (KGFR/FGFR2b) is a major paracrine mediator of epithelial homeostasis and appears to have an unique and unusual role in epithelial tissues, exerting a tumor-suppressive function in vitro and in vivo. With the aim to better elucidate the molecular events involved in the pathological activity of 16E5, we investigated if the viral protein would be able to affect the KGFR expression, signaling and turnover by interference with its degradative and recycling endocytic pathways. Quantitative reverse transcriptase-PCR and biochemical approaches on human keratinocytes transfected with 16E5-HA showed that E5 protein is able to induce KGFR down-modulation at both transcript and protein levels. Immunofluorescence microscopy in double transfected cells expressing both E5 and KGFR revealed that the viral protein alters the receptor endocytic trafficking and triggers its endosomal sorting to the indirect juxtanuclear recycling pathway. The shift from lysosomal degradation to recycling at the plasma membrane correlates with a reduced phosphorylation of the fibroblast growth factor receptor substrate-2alpha tyrosine 196, the major docking site for Grb2-Cbl complexes responsible for receptor ubiquitination and degradation. 5'-Bromo-deoxyuridine incorporation assay demonstrated that expression of 16E5 induces a decrease in the growth response to the receptor ligands as a consequence of KGFR down-modulation, suggesting that 16E5 might have a role on HPV infection in perturbing the KGFR-mediated physiological behavior of confluent keratinocytes committed to differentiation. PMID- 21625214 TI - The Ron receptor promotes prostate tumor growth in the TRAMP mouse model. AB - The Ron receptor tyrosine kinase (TK) is overexpressed in many cancers, including prostate cancer. To examine the significance of Ron in prostate cancer in vivo, we utilized a genetically engineered mouse model, referred to as TRAMP mice, that is predisposed to develop prostate tumors. In this model, we show that prostate tumors from 30-week-old TRAMP mice have increased Ron expression compared with age-matched wild-type prostates. Based on the upregulation of Ron in human prostate cancers and in this murine model of prostate tumorigenesis, we hypothesized that this receptor has a functional role in the development of prostate tumors. To test this hypothesis, we crossed TRAMP mice with mice that are deficient in Ron signaling (TK-/-). Interestingly, TK-/- TRAMP+ mice show a significant decrease in prostate tumor mass relative to TRAMP mice containing functional Ron. Moreover, TK-/- TRAMP+ prostate tumors exhibited decreased tumor vascularization relative to TK+/+ TRAMP+ prostate tumors, which correlated with reduced levels of the angiogenic molecules vascular endothelial growth factor and CXCL2. Although Ron loss did not alter tumor cell proliferation, a significant decrease in cell survival was observed. Similarly, murine prostate cancer cell lines containing a Ron deficiency exhibited decreased levels of active nuclear factor-kappaB, suggesting that Ron may be important in regulating prostate cell survival at least partly through this pathway. In total, our data show for the first time that Ron promotes prostate tumor growth, prostate tumor angiogenesis and prostate cancer cell survival in vivo. PMID- 21625215 TI - microRNA-29 can regulate expression of the long non-coding RNA gene MEG3 in hepatocellular cancer. AB - The human genome is replete with long non-coding RNAs (lncRNA), many of which are transcribed and likely to have a functional role. Microarray analysis of >23,000 lncRNAs revealed downregulation of 712 (~3%) lncRNA in malignant hepatocytes, among which maternally expressed gene 3 (MEG3) was downregulated by 210-fold relative to expression in non-malignant hepatocytes. MEG3 expression was markedly reduced in four human hepatocellular cancer (HCC) cell lines compared with normal hepatocytes by real-time PCR. RNA in situ hybridization showed intense cytoplasmic expression of MEG3 in non-neoplastic liver with absent or very weak expression in HCC tissues. Enforced expression of MEG3 in HCC cells significantly decreased both anchorage-dependent and -independent cell growth, and induced apoptosis. MEG3 promoter hypermethylation was identified by methylation-specific PCR and MEG3 expression was increased with inhibition of methylation with either 5-Aza-2-Deoxycytidine, or siRNA to DNA Methyltransferase (DNMT) 1 and 3b in HCC cells. MiRNA-dependent regulation of MEG3 expression was studied by evaluating the involvement of miR-29, which can modulate DNMT 1 and 3. Overexpression of mir 29a increased expression of MEG3. GTL2, the murine homolog of MEG3, was reduced in liver tissues from hepatocyte-specific miR-29a/b1 knock-out mice compared with wild-type controls. These data show that methylation-dependent tissue-specific regulation of the lncRNA MEG3 by miR-29a may contribute to HCC growth and highlight the inter-relationship between two classes of non-coding RNA, miRNAs and lncRNAs, and epigenetic regulation of gene expression. PMID- 21625216 TI - EPLIN downregulation promotes epithelial-mesenchymal transition in prostate cancer cells and correlates with clinical lymph node metastasis. AB - Epithelial-mesenchymal transition (EMT) is a crucial mechanism for the acquisition of migratory and invasive capabilities by epithelial cancer cells. By conducting quantitative proteomics in experimental models of human prostate cancer (PCa) metastasis, we observed strikingly decreased expression of EPLIN (epithelial protein lost in neoplasm; or LIM domain and actin binding 1, LIMA-1) upon EMT. Biochemical and functional analyses demonstrated that EPLIN is a negative regulator of EMT and invasiveness in PCa cells. EPLIN depletion resulted in the disassembly of adherens junctions, structurally distinct actin remodeling and activation of beta-catenin signaling. Microarray expression analysis identified a subset of putative EPLIN target genes associated with EMT, invasion and metastasis. By immunohistochemistry, EPLIN downregulation was also demonstrated in lymph node metastases of human solid tumors including PCa, breast cancer, colorectal cancer and squamous cell carcinoma of the head and neck. This study reveals a novel molecular mechanism for converting cancer cells into a highly invasive and malignant form, and has important implications in prognosis and treating metastasis at early stages. PMID- 21625217 TI - Mitochondrial respiratory chain complexes: apoptosis sensors mutated in cancer? AB - Mutations in cancer cells affecting subunits of the respiratory chain (RC) indicate a central role of oxidative phosphorylation for tumourigenesis. Recent studies have suggested that such mutations of RC complexes impact apoptosis induction. We review here the evidence for this hypothesis, which in particular emerged from work on how complex I and II mediate signals for apoptosis. Both protein aggregates are specifically inhibited for apoptosis induction through different means by exploiting with protease activation and pH change, two widespread but independent features of dying cells. Nevertheless, both converge on forming reactive oxygen species for the demise of the cell. Investigations into these mitochondrial processes will remain a rewarding area for unravelling the causes of tumourigenesis and for discovering interference options. PMID- 21625218 TI - Hypoxia-driven cell motility reflects the interplay between JMY and HIF-1alpha. AB - Junction-mediating and regulatory protein (JMY) is a novel p53 cofactor that regulates p53 activity during stress. JMY interacts with p300/CBP, which are ubiquitous transcriptional co-activators that interact with a variety of sequence specific transcription factors, including hypoxia-inducible factor-1alpha (HIF 1alpha). In addition, JMY is an actin-nucleating protein, which, through its WH2 domains, stimulates cell motility. In this study, we show that JMY is upregulated during hypoxia in a HIF-1alpha-dependent manner. The JMY gene contains HIF responsive elements in its promoter region and HIF-1alpha is recruited to its promoter during hypoxia. HIF-1alpha drives transcription of JMY, which accounts for its induction under hypoxia. Moreover, the enhanced cell motility and invasion that occurs during hypoxia requires JMY, as depleting JMY under hypoxic conditions causes decreased cell motility. Our results establish the interplay between JMY and HIF-1alpha as a new mechanism that controls cell motility under hypoxic stress. PMID- 21625219 TI - Cancer cells promote survival through depletion of the von Hippel-Lindau tumor suppressor by protein crosslinking. AB - Nuclear factor-kappaB (NF-kappaB) and insulin-like growth factor-1 (IGF-1) mediated signaling is associated with different tumors including renal cell carcinoma. NF-kappaB- and IGF-1-mediated signaling is found to be inhibited in the presence of wild-type von Hippel-Lindau (VHL) tumor suppresser gene. Therefore, negative regulator of VHL may be a good target for regulating NF kappaB and IGF-1R. In this study, we found that VHL, a tumor suppressor protein that downregulates the NF-kappaB activity and the stability of IGF-1R was depleted by TGase 2 through polymerization via crosslinking and proteasomal degradation in kidney, breast and ovary cancer cell lines. We also found that TGase 2 knockdown promotes hypoxia-inducible factor 1alpha (HIF-1alpha) degradation, and thereby decrease HIF-1alpha transcriptional activity. Importantly, VHL expression was decreased in vivo in TGase-2-transgenic mice, and this was associated with increased NF-kappaB activity and the levels of expression of IGF-1R, HIF-1alpha and erythropoietin in kidney tissue. These results suggest a novel mechanism of regulation of the VHL tumor suppressor by TGase 2 that appears to be independent of the known cancer regulatory mechanisms. PMID- 21625220 TI - Overexpression of GalNAc-transferase GalNAc-T3 promotes pancreatic cancer cell growth. AB - O-linked glycans of secreted and membrane-bound proteins have an important role in the pathogenesis of pancreatic cancer by modulating immune responses, inflammation and tumorigenesis. A critical aspect of O-glycosylation, the position at which proteins are glycosylated with N-acetyl-galactosamine on serine and threonine residues, is regulated by the substrate specificity of UDP GalNAc:polypeptide N-acetylgalactosaminyl-transferases (GalNAc-Ts). Thus, GalNAc Ts regulate the first committed step in O-glycosylated protein biosynthesis, determine sites of O-glycosylation on proteins and are important for understanding normal and carcinoma-associated O-glycosylation. We have found that one of these enzymes, GalNAc-T3, is overexpressed in human pancreatic cancer tissues and suppression of GalNAc-T3 significantly attenuates the growth of pancreatic cancer cells in vitro and in vivo. In addition, suppression of GalNAc T3 induces apoptosis of pancreatic cancer cells. Our results indicate that GalNAc T3 is likely involved in pancreatic carcinogenesis. Modification of cellular glycosylation occurs in nearly all types of cancer as a result of alterations in the expression levels of glycosyltransferases. We report guanine the nucleotide binding protein, alpha-transducing activity polypeptide-1 (GNAT1) as a possible substrate protein of GalNAc-T3. GalNAc-T3 is associated with O-glycosylation of GNAT1 and affects the subcellular distribution of GNAT1. Knocking down endogenous GNAT1 significantly suppresses the growth/survival of PDAC cells. Our results imply that GalNAc-T3 contributes to the function of O-glycosylated proteins and thereby affects the growth and survival of pancreatic cancer cells. Thus, substrate proteins of GalNAc-T3 should serve as important therapeutic targets for pancreatic cancers. PMID- 21625221 TI - NF-kappaB suppresses ROS levels in BCR-ABL(+) cells to prevent activation of JNK and cell death. AB - Elevated levels of reactive oxygen species (ROS) are found in most oncogenically transformed cells and are proposed to promote cellular transformation through mechanisms such as inhibition of phosphatases. BCR-ABL, the oncoprotein associated with the majority of chronic myeloid leukemias (CMLs), induces accumulation of intracellular ROS, causing enhanced signaling downstream of PI3K. Previously we have shown that the transcription factor nuclear factor-kappa B (NF kappaB) is activated by BCR-ABL expression and is required for BCR-ABL-mediated cellular transformation. Inhibition of IkappaB kinase (IKKbeta) and NF-kappaB leads to cell death through an unknown mechanism. Here, we analyze the potential involvement of NF-kappaB in moderating BCR-ABL-induced ROS levels to protect from death. The data confirm that BCR-ABL promotes ROS and demonstrate that NF-kappaB prevents excessive levels. Inhibition of NF-kappaB leads to an increase in ROS levels and to cell death, which is at least partially controlled through ROS induced c-Jun N-terminal kinase activity. The data demonstrate that one function for NF-kappaB in oncogenesis is the suppression of oncoprotein-induced ROS levels and that inhibition of NF-kappaB in some cancers, including CML, will increase ROS levels and promote cell death. PMID- 21625222 TI - ABCG2 is a direct transcriptional target of hedgehog signaling and involved in stroma-induced drug tolerance in diffuse large B-cell lymphoma. AB - Successful treatment of diffuse large B-cell lymphoma (DLBCL) is frequently hindered by the development of resistance to conventional chemotherapy resulting in disease relapse and high mortality. High expression of antiapoptotic and/or drug transporter proteins induced by oncogenic signaling pathways has been implicated in the development of chemoresistance in cancer. Previously, our studies showed that high expression of adenosine triphosphate-binding cassette drug transporter ABCG2 in DLBCL correlated inversely with disease- and failure free survival. In this study, we have implicated activated hedgehog (Hh) signaling pathway as a key factor behind high ABCG2 expression in DLBCL through direct upregulation of ABCG2 gene transcription. We have identified a single binding site for GLI transcription factors in the ABCG2 promoter and established its functionality using luciferase reporter, site-directed mutagenesis and chromatin-immunoprecipitation assays. Furthermore, in DLBCL tumor samples, significantly high ABCG2 and GLI1 levels were found in DLBCL tumors with lymph node involvement in comparison with DLBCL tumor cells collected from pleural and/or peritoneal effusions. This suggests a role for the stromal microenvironment in maintaining high levels of ABCG2 and GLI1. Accordingly, in vitro co-culture of DLBCL cells with HS-5 stromal cells increased ABCG2 mRNA and protein levels by paracrine activation of Hh signaling. In addition to ABCG2, co culture of DLBCL cells with HS-5 cells also resulted in increase expression of the antiapoptotic proteins BCL2, BCL-xL and BCL2A1 and in induced chemotolerance to doxorubicin and methotrexate, drugs routinely used for the treatment of DLBCL. Similarly, activation of Hh signaling in DLBCL cell lines with recombinant Shh N terminal peptide resulted in increased expression of BCL2 and ABCG2 associated with increased chemotolerance. Finally, functional inhibition of ABCG2 drug efflux activity with fumitremorgin C or inhibition of Hh signaling with cyclopamine-KAAD abrogated the stroma-induced chemotolerance suggesting that targeting ABCG2 and Hh signaling may have therapeutic value in overcoming chemoresistance in DLBCL. PMID- 21625223 TI - Tp53 deletion in B lineage cells predisposes mice to lymphomas with oncogenic translocations. AB - Inactivating Tp53 mutations are frequent genetic lesions in human tumors that harbor genomic instability, including B lineage lymphomas with IG translocations. Antigen receptor genes are assembled and modified in developing lymphocytes by RAG/AID-initiated genomic rearrangements that involve the induction of DNA double strand breaks (DSBs). Although TP53 inhibits the persistence of DSBs and induces apoptosis to protect cells from genomic instability and transformation, the development of spontaneous tumors harboring clonal translocations has not been reported in mice that only lack wild-type Tp53 protein or express Tp53 mutants. Tp53-deficient (Tp53(-/-)) mice succumb to T lineage lymphomas lacking clonal translocations but develop B lymphoid tumors containing immunoglobulin (Ig) translocations upon combined inactivation of DSB repair factors, RAG mutation or AID overexpression; mice expressing apoptosis-defective Tp53 mutants develop B cell lymphomas that have not been characterized for potential genomic instability. As somatic rather than germline inactivating mutations of TP53 are typically associated with human cancers and Tp53 deletion has cellular context dependent effects upon lymphocyte transformation, we generated mice with conditional Tp53 deletion in lineage-committed B lymphocytes to avoid complications associated with defective Tp53 responses during embryogenesis and/or in multi-lineage potential cells and, thereby, directly evaluate the potential physiological role of Tp53 in suppressing translocations in differentiated cells. These mb1-cre:Tp53(flox/flox) mice succumbed to lymphoid tumors containing Ig gene rearrangements and immunophenotypes characteristic of B cells from various developmental stages. Most mb1-cre:Tp53(flox/flox) tumors harbored clonal translocations, including Igh/c-myc or other oncogenic translocations generated by the aberrant repair of RAG/AID-generated DSBs. Our data indicate that Tp53 serves critical functions in B lineage lymphocytes to prevent transformation caused by translocations in cell populations experiencing physiological levels of RAG/AID-initiated DSB intermediates, and provide evidence that the somatic TP53 mutations found in diffuse large B-cell lymphoma and Burkitt's lymphoma may contribute to the development of these human malignancies. PMID- 21625225 TI - Assessing causal relationships between treatments and clinical outcomes: always read the fine print. AB - Changes in clinical practice should be driven by relevant and reliable evidence. Hence, adoption of a new therapy requires demonstrating that it provides (causes) benefit. Such evidence is generally obtained from intent-to-treat analyses of randomized clinical trials (RCTs). In this paper, we review other approaches to assessing the causal relationship between treatments and outcomes: (1) inference from non-randomized (observational) studies, (2) analysis of randomized studies where patients received treatments other than those to which they were randomized and (3) analysis of studies where the outcome of interest is sometimes unobservable because of a competing event (competing risks). We conclude that for the practice-changing demonstration of a favorable benefit-to-risk ratio, the gold standard is the intent-to-treat analysis of RCTs. At the same time, we illustrate how careful application of special statistical methods for assessment of treatment-outcome causation can be instrumental in complementing existing randomized evidence and guiding design of future research. PMID- 21625224 TI - Proposed definition of 'poor mobilizer' in lymphoma and multiple myeloma: an analytic hierarchy process by ad hoc working group Gruppo ItalianoTrapianto di Midollo Osseo. AB - Many lymphoma and myeloma patients fail to undergo ASCT owing to poor mobilization. Identification of poor mobilizers (PMs) would provide a tool for early intervention with new mobilization agents. The Gruppo italianoTrapianto di Midollo Osseo working group proposed a definition of PMs applicable to clinical trials and clinical practice. The analytic hierarchy process, a method for group decision making, was used in setting prioritized criteria. Lymphoma or myeloma patients were defined as 'proven PM' when: (1) after adequate mobilization (G-CSF 10 MUg/kg if used alone or >=5 MUg/kg after chemotherapy) circulating CD34(+) cell peak is <20/MUL up to 6 days after mobilization with G-CSF or up to 20 days after chemotherapy and G-CSF or (2) they yielded <2.0 * 10(6) CD34(+) cells per kg in <=3 apheresis. Patients were defined as predicted PMs if: (1) they failed a previous collection attempt (not otherwise specified); (2) they previously received extensive radiotherapy or full courses of therapy affecting SC mobilization; and (3) they met two of the following criteria: advanced disease (>=2 lines of chemotherapy), refractory disease, extensive BM involvement or cellularity <30% at the time of mobilization; age >=65 years. This definition of proven and predicted PMs should be validated in clinical trials and common clinical practice. PMID- 21625226 TI - Testicular function in patients with type 1 diabetes treated with high-dose CY and autologous hematopoietic SCT. PMID- 21625227 TI - Clinical features of calcineurin inhibitor-induced pain syndrome after allo-SCT. PMID- 21625228 TI - Candida kefyr fungal enteritis following autologous BMT. PMID- 21625229 TI - Interferon consensus sequence binding protein-induced cell proliferation is mediated by TGF-beta signaling and p38 MAPK activation. AB - Interferon consensus sequence binding protein (ICSBP), also known as interferon regulatory factor (IRF)-8, is a member of the interferon (IFN)-gamma regulatory transcription factors. Studies have suggested a connection between TGF-beta signaling and IRFs. Thus, we investigated the effect of ICSBP on transforming growth factor (TGF)-beta signaling in HL-60, an acute promyelocytic leukemia cell line. Stable expression of ICSBP in HL-60 cells resulted in strong induction of TGF-beta receptor expression and activation of non-Smad as well as Smad pathways. ICSBP expression also augmented cell growth. ICSBP knockdown with small interfering RNA (siRNA) attenuated cell growth and decreased TGF-beta receptor I (TGF-betaRI) expression. In addition, reduction of TGF-betaRI using siRNA or pharmacological inhibitor reduced growth of ICSBP-expressing cells. ICSBP expression also led to increased phosphorylation and activation of Akt and p38 MAPK. However, p38 MAPK, but not PI3K-Akt, inhibition abrogated ICSBP-mediated proliferation. Furthermore, siRNA knockdown of either ICSBP or TGF-betaRI resulted in decreased p38 activation. Intriguingly, TGF-beta-activated kinase 1 (TAK-1), which phosphorylates p38, was activated in ICSBP-expressing cells and its activity was reduced by TGF-betaRI inhibition. Finally, siRNA knockdown of ICSBP or TGF-betaRI reduced TAK-1 phosphorylation. This study identifies a novel role for ICSBP in regulating cell growth via TGF-beta receptor upregulation and subsequent activation of the TGF-beta receptor/TAK-1/p38 pathway. PMID- 21625230 TI - Neuroimaging as endpoints in clinical trials: are we there yet? Perspective from the first Provence workshop. PMID- 21625231 TI - Deep brain stimulation and the role of astrocytes. AB - Deep brain stimulation (DBS) has emerged as a powerful surgical therapy for the management of treatment-resistant movement disorders, epilepsy and neuropsychiatric disorders. Although DBS may be clinically effective in many cases, its mode of action is still elusive. It is unclear which neural cell types are involved in the mechanism of DBS, and how high-frequency stimulation of these cells may lead to alleviation of the clinical symptoms. Neurons have commonly been a main focus in the many theories explaining the working mechanism of DBS. Recent data, however, demonstrates that astrocytes may be active players in the DBS mechanism of action. In this review article, we will discuss the potential role of reactive and neurogenic astrocytes (neural progenitors) in DBS. PMID- 21625232 TI - An eight-gene expression signature for the prediction of survival and time to treatment in chronic lymphocytic leukemia. PMID- 21625233 TI - Tumor suppressor TNFAIP3 (A20) is frequently deleted in Sezary syndrome. AB - Despite recent therapeutic improvements, the prognosis for patients suffering from Sezary syndrome (SS), a disseminated form of cutaneous T-cell lymphomas, is still poor. We identified bi- and monoallelic deletions of the tumor necrosis factor-alpha-induced protein 3 gene (TNFAIP3; A20) in a high proportion of SS patients as well as biallelic A20 deletion in the SS-derived cell line SeAx. Furthermore, we demonstrate that inhibition of A20 activates the NF-kappaB pathway thereby increasing the proliferation of normal T lymphocytes. On the other hand, the reconstitution of A20 expression slowed down the cell cycle in SeAx cells. Recently A20 inactivation has been reported in various B-cell lymphomas. In this study, we show that A20 is also a putative tumor suppressor in the T-cell malignancy-SS. PMID- 21625234 TI - Genomic 5-hydroxymethylcytosine levels correlate with TET2 mutations and a distinct global gene expression pattern in secondary acute myeloid leukemia. PMID- 21625235 TI - A phase 1 trial dose-escalation study of tipifarnib on a week-on, week-off schedule in relapsed, refractory or high-risk myeloid leukemia. AB - Inhibition of farnesyltransferase (FT) activity has been associated with in vitro and in vivo anti-leukemia activity. We report the results of a phase 1 dose escalation study of tipifarnib, an oral FT inhibitor, in patients with relapsed, refractory or newly diagnosed (if over age 70) acute myelogenous leukemia (AML), on a week-on, week-off schedule. Forty-four patients were enrolled, two patients were newly diagnosed, and the rest were relapsed or refractory to previous treatment, with a median age of 61 (range 33-79). The maximum tolerated dose was determined to be 1200 mg given orally twice daily (b.i.d.) on this schedule. Cycle 1 dose-limiting toxicities were hepatic and renal. There were three complete remissions seen, two at the 1200 mg b.i.d. dose and one at the 1000 mg b.i.d. dose, with minor responses seen at the 1400 mg b.i.d. dose level. Pharmacokinetic studies performed at doses of 1400 mg b.i.d. showed linear behavior with minimal accumulation between days 1-5. Tipifarnib administered on a week-on, week-off schedule shows activity at higher doses, and represents an option for future clinical trials in AML. PMID- 21625236 TI - Redundancy and specificity of the metalloprotease system mediating oncogenic NOTCH1 activation in T-ALL. AB - Oncogenic mutations in NOTCH1 are present in over 50% of T-cell lymphoblastic leukemias (T-ALLs). Activation of NOTCH1 requires a double proteolytic processing in the extracellular region of the receptor (S2) and in the transmembrane domain (S3). Currently, anti-NOTCH1 therapies based on the inhibition of S3 processing via small molecule gamma-secretase inhibitors are in development. Here we report on the characterization of the protease system responsible for S2 processing of NOTCH1 in T-ALL. Analysis of NOTCH1 heterodimerization (HD) class I, NOTCH1 HD class II and NOTCH1 JME alleles characterized by increased and aberrant S2 processing shows that both ADAM10 (a disintegrin and metalloprotease 10), a metalloprotease previously implicated in activation of wild-type NOTCH1 in mammalian cells, and ADAM17, a closely related protease capable of processing NOTCH1 in vitro, contribute to the activation of oncogenic forms of NOTCH1. However, and despite this apparent functional redundancy, inhibition of ADAM10 is sufficient to blunt NOTCH1 signaling in T-ALL lymphoblasts. These results provide further insight on the mechanisms that control the activation of oncogenic NOTCH1 mutants and identify ADAM10 as potential therapeutic target for the inhibition of oncogenic NOTCH1 in T-ALL. PMID- 21625237 TI - A novel ABL1 fusion to the SH2 containing inositol phosphatase-1 (SHIP1) in acute lymphoblastic leukemia (ALL). PMID- 21625238 TI - Evidence-based management of upper tract urolithiasis in the spinal cord-injured patient. AB - OBJECTIVE: The objective is to review the published literature on the aetiology and evidence-based management of stone disease in the spinal cord-injured patient. METHODS: A PubMed and Medline search was performed using the terms 'spinal cord injury', 'paraplegia', 'stone', 'nephrolithiasis', 'urolithiasis', 'calculus', 'spinal cord injury' or 'paraplegia' with 'SWL', 'ureteroscopy', 'chemolysis' and 'PCNL.' The Cochrane database, the National Institute for Clinical Excellence guidelines and the Scottish Intercollegiate guidelines were searched using the terms 'spinal cord injury' and 'urolithiasis' and 'nephrolithiasis'. RESULTS: A total of 32 papers were identified, mainly case series or case-cohort studies with few contemporary papers. The risk of developing a renal stone after spinal cord injury (SCI) is between 7 and 20% over a period of 8-10 years. Stone formation may be related to early demineralisation of bone or chronic infection. Biochemical abnormalities are not significantly different between stone-forming and non-stone forming patients, though these patients differ from healthy controls. Presentation may be atypical, but is most commonly recurrent urinary tract infection. Treatment may be complicated by lower limb contractures limiting retrograde access. Several case series report success with shock wave lithotripsy varying from 50 to 70%, though comparisons are limited by heterogeneous indications and reporting. Percutaneous nephrolithotomy remains the gold standard for stones measuring 2 cm and above. Stone-free rates of 90% have been reported, though surgery was often complex with higher complication rates. CONCLUSION: Management of upper urinary tract stones in patients with SCI is complex regarding surgical technique, post-operative care and recurrence rates. Further contemporary case series must use standardised reporting tools to allow valid comparisons. PMID- 21625239 TI - Locomotor training using a robotic device in patients with subacute spinal cord injury. AB - STUDY DESIGN: Quasi experiment; single experimental group with matched historical control. OBJECTIVES: To evaluate the effect of an additive robotic-assisted gait training (RAGT) using the Lokomat system on the neurological and functional outcomes of patients with subacute spinal cord injury (SCI). SETTING: Department of Physical Medicine and Rehabilitation. METHODS: A total of 28 subacute SCI patients were treated by RAGT, 2-3 times a week, 30-45 min every treatment, concomitantly with regular physiotherapy. As control, for each patient, we matched a comparable patient treated in the same department in previous years, according to age, severity of injury, level of injury and cause. The main outcomes were: the AIS (American Spinal Injury Association impairment scale) the spinal cord independence measurement (SCIM) score, the walking index for SCI II (WISCI II) and functional ambulation category scale (FAC). RESULTS: At the end of rehabilitation, both groups showed a significant improvement in both the FAC score and the WISCI score (P<0.01) without differences between the groups. Functional abilities, according to the SCIM score, were also improved, with a significant interaction effect; the RAGT patients improve by 30+/-20 points, which was significantly greater gain as compared with the controls, 21+/-14 points (P=0.05). This improvement was mainly due to the change in the SCIM motor subscales. CONCLUSION: RAGT is an important additional treatment to improve the functional outcome of subacute SCI patients. Larger, controlled studies are still required to determine the optimal timing and protocol design for the maximal efficacy of RAGT in SCI patients. PMID- 21625240 TI - Serum hsCRP and visfatin are elevated and correlate to carotid arterial stiffness in spinal cord-injured subjects. AB - STUDY DESIGN: Cross-sectional comparison, control group. OBJECTIVES: To investigate the relationship between carotid arterial stiffness and circulating markers for cardiovascular disease (CVD) in spinal cord-injured (SCI) subjects compared with able-bodied (AB) individuals. SETTING: University Research Laboratory, University of Louisville. METHODS: SCI (n=14) and AB (n=13) subjects between 20-52 years of age were recruited to participate in the study. B-mode Doppler ultrasound was used to obtain carotid artery diameter measurements. Arterial stiffness was assessed via the stiffness index and distensibility coefficient. Markers of CVD risk were obtained by fasting blood draw. RESULTS: Carotid arterial stiffness index (P=0.061) and distensibility coefficient (P=0.370) were not different between the SCI and AB groups. The SCI group had higher high-sensitivity C-reactive protein (hsCRP) (P=0.046), triglycerides (P=0.017), leptin (P=0.040) and visfatin (P<0.001) compared with the control group. Visfatin (r=0.559, P=0.047), hsCRP (r=0.633, P=0.037), insulin (r=0.637, P=0.019) and HOMA (r=0.614, P=0.026) significantly correlated with carotid arterial stiffness index in the SCI group. CONCLUSION: This study demonstrated that SCI subjects are at a high cardiovascular risk as indicated by elevated hsCRP levels. Elevations in hsCRP and visfatin may contribute to accelerated atherogenic processes in the SCI population. PMID- 21625241 TI - Spinal cord injury sequelae alter drug pharmacokinetics: an overview. AB - STUDY DESIGN: Literature review. OBJECTIVES: Critical review of the literature published on the physiological alterations caused by spinal cord injury (SCI) and their effect on the pharmacokinetic parameters of commonly employed drugs. The review introduces the most recent treatment protocols of a variety of drugs, enabling the modern clinician to apply efficacious and cost-effective solutions to the pharmacological treatment of SCI patients. METHODS: Studies published in international indexed journals up to January 2011 were selected from the PubMed database. RESULTS: The review evaluated the sequelae of SCI and their effect on pharmacological processes. The results demonstrated that these alterations affected the pharmacokinetics of drugs commonly administered to SCI patients, such as antibiotics, muscle relaxants, immunosuppressants and analgesics. CONCLUSION: There are multiple etiologies to SCI and patients present with varying degrees of impairment. Factors such as level of injury and completeness of the injury create a very heterogeneous population within the SCI community. The heterogeneity of this population creates a problem when trying to standardize pharmacokinetic (PK) parameters. It is because of this that specific physiological alterations must be linked to changes in PK and be identified within the clinical setting. This relationship between physiology and PK enables the clinician to be alert for possible pharmacological complications in individual patients based on their clinical manifestations. Future research should aim to develop rigorous therapeutic guidelines tailored to the diverse manifestations of SCI so as to provide effective, affordable and safe pharmacotherapy. PMID- 21625242 TI - ToCUEST: a task-oriented client-centered training module to improve upper extremity skilled performance in cervical spinal cord-injured persons. AB - OBJECTIVES: Given (a) the importance of upper extremity skill performance in persons with cervical spinal cord injury (C-SCI), (b) the limited number of studies on arm hand functioning in C-SCI and (c) the discrepancy between the importance of client centeredness and the lack of a theoretical framework and practical guidelines on how to apply client centeredness into practice, the aim of the present study is to (1) develop a training concept in which rehabilitation of the upper extremity focuses on patient's needs and wishes and (2) to put this concept into practice for tetraplegic persons. METHODS: The training concept contains two main components. The client-centered component includes both extracting individual goals and defining and objectifying these goals. The task oriented component incorporates a task analysis and the composition of an individual training program. RESULTS: A task-oriented client-centered training aimed at improving upper extremity skilled performance in tetraplegic persons is described step by step. The Canadian Occupational Performance Measure is used to identify patient's specific needs. The Goal Attainment Scale is applied to define and objectify individual goals. A task analysis is essential to map limiting and facilitating factors of the person, the task and the environment and to identify which aspects should be trained. An individual task-oriented training program is composed, based on principles of motor learning and training physiology and the use of assistive devices. Each step is explained and illustrated using a case example. PMID- 21625243 TI - Stimulation of defecation in spinal cord-injured rats by a centrally acting ghrelin receptor agonist. AB - STUDY DESIGN: Animal proof of principle study. OBJECTIVES: To determine whether capromorelin, a compound that causes defecation by stimulating ghrelin receptors within the lumbosacral defecation centers, is effective after spinal cord injury (SCI), and whether SCI significantly alters sensitivity to the compound. SETTING: University of Melbourne and Austin Hospital, Melbourne, Australia. METHODS: Rats were subjected to spinal cord contusion injury or were sham-operated. At 6 weeks after surgery, effects of capromorelin on blood pressure, heart rate and propulsive contractions of the colorectum were investigated. RESULTS: Capromorelin caused robust propulsive activity in the colorectum soon after its application. The compound was similarly effective in naive, sham-operated and spinal cord-injured rats. Blood pressure increases caused by capromorelin were not exaggerated after SCI, and there was no evidence of phasic blood pressure increases when the colon was contracted by the compound. CONCLUSION: Capromorelin is a therapeutic compound that could potentially be used to relieve constipation by triggering defecation in spinal cord-injured patients. PMID- 21625245 TI - Microbial ecology: Bacteria reinforce plant defences. PMID- 21625246 TI - Immunology: NLRP6 keeps the bad bacteria at bay. PMID- 21625248 TI - Microbial carbon pump: additional considerations. PMID- 21625249 TI - Parasitology: Adding insult to injury. PMID- 21625250 TI - Regulation of mucosal mast cell activation by short interfering RNAs targeting syntaxin4. AB - Mucosal mast cells (MMCs) have an important role in allergic inflammation, and effective antagonists are required for their regulation. To discover a possible mechanism of controlling the activation of MMCs, we investigated the expression and function of syntaxin4, one of the soluble membrane N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins, in RBL-2H3 cells, which is a rat mucosal mast cell line. Syntaxin4 silencing was induced by transfection of short interfering RNAs (siRNAs). Syntaxin4 was knocked down in mast cells at both the mRNA and protein levels. The release of granule contents that are involved in inflammation, such as histamine and hexosaminidase, was significantly suppressed by the gene silencing of syntaxin4. Silencing of this gene was also induced in the trachea and bronchi of rats by intratracheal application of the siRNAs using an atelocollagen delivery system. The activation of MMCs, which was monitored by the level of rat mast cell protease-II (RMCPII) in the bronchoalveolar lavage fluid (BALF), was inhibited, and asthmatic airway constriction was prevented by administration of the syntaxin/atelocollagen complex. These results indicate that siRNAs targeting syntaxin4 can stabilize mucosal mast cells and may have beneficial therapeutic effects on the asthmatic response. PMID- 21625251 TI - Sclerodermatous chronic graft-versus-host disease induced by host T-cell-mediated autoimmunity. AB - Despite a long-standing hypothesis that chronic graft-versus-host disease (cGVHD) is an autoimmune disorder, most mouse models of cGVHD have been developed on the assumption that donor T cells are essential for its development. Here we show that cGVHD may be caused by autoreactive host T cells in mice that have been lethally irradiated and grafted with T-cell-depleted allogeneic bone marrow cells. In this chimera, host T cells derived from radioresistant intrathymic T cell precursors caused dermal fibrosis and periportal inflammation, without the requirement for donor T cells. The lack of host DCs within the thymus after high dose irradiation allowed autoreactive host T cells to escape thymic negative selection. Moreover, the homeostatic expansion of these T cells may augment their autoreactivity. These findings indicate that host T-cell-mediated cGVHD is an autoimmune process that occurs following the grafting of T-cell-depleted BM cells into hosts with functioning thymuses. We propose, based on the present data, that host T-cell-dependent autoimmunity is a potential mechanism by which cGVHD is induced. PMID- 21625252 TI - Cytidine deaminase single-nucleotide polymorphism is predictive of toxicity from gemcitabine in patients with pancreatic cancer: RTOG 9704. AB - The aim of this study is to validate the prognostic and predictive value of the non-synonymous cytidine deaminase (CDA) Lys27Gln polymorphism for hematological toxicity and survival using a randomized phase III adjuvant trial (Radiation Therapy Oncology Group (RTOG) 9704) in pancreatic cancer in which one treatment arm received gemcitabine. CDA is involved in gemcitabine inactivation, and there is conflicting data on the role of the non-synonymous CDA Lys27Gln polymorphism in predicting toxicity and survival in cancer patients treated with gemcitabine. RTOG 9704 randomized 538 patients after pancreatic resection to receive radiotherapy with either 5-fluorouracil (5-FU) or gemcitabine. CDA Lys27Gln polymorphism genotype was analyzed. We tested an association between CDA single nucleotide polymorphism genotype and the survival outcome by the Cox proportional hazard model adjusting for other covariates, as well as toxicity by the logistic regression model. There is statistically significant more severe hematological toxicity in patients treated with gemcitabine with either the homozygote wild type genotype (Lys/Lys) alone (odds ratio (OR)=0.06, P=0.01), or in combination with the heterozygote (Lys/Gln; OR=0.14, P=0.03) when compared with homozygote variant genotype (Gln/Gln) when adjusted for other covariates. This was not seen in the non-gemcitabine treated arm. There are no genotype differences with respect to survival outcome. In conclusion, in this prospective randomized adjuvant study of patients with pancreatic cancer, the CDA Lys27Gln polymorphism is validated as a predictive marker of gemcitabine hematological toxicity, but not with treatment response or survival. PMID- 21625253 TI - An update on ABCB1 pharmacogenetics: insights from a 3D model into the location and evolutionary conservation of residues corresponding to SNPs associated with drug pharmacokinetics. AB - The human ABCB1 protein, (P-glycoprotein or MDR1) is a membrane-bound glycoprotein that harnesses the energy of ATP hydrolysis to drive the unidirectional transport of substrates from the cytoplasm to the extracellular space. As a large range of therapeutic agents are known substrates of ABCB1 protein, its role in the onset of multidrug resistance has been the focus of much research. This role has been of particular interest in the field of pharmacogenomics where genetic variation within the ABCB1 gene, particularly in the form of single nucleotide polymorphisms (SNPs), is believed to contribute to inter-individual variation in ABCB1 function and drug response. In this review we provide an update on the influence of coding region SNPs within the ABCB1 gene on drug pharmacokinetics. By utilizing the crystal structure of the mouse ABCB1 homolog (Abcb1a), which is 87% homologous to the human sequence, we accompany this discussion with a graphical representation of residue location for amino acids corresponding to human ABCB1 coding region SNPs. Also, an assessment of residue conservation, which is calculated following multiple sequence alignment of 11 confirmed sequences of ABCB1 homologs, is presented and discussed. Superimposing a 'heat map' of residue homology to the Abcb1a crystal structure has permitted additional insights into both the conservation of individual residues and the conservation of their immediate surroundings. Such graphical representation of residue location and conservation supplements this update of ABCB1 pharmacogenetics to help clarify the often confounding reports on the influence of ABCB1 polymorphisms on drug pharmacokinetics and response. PMID- 21625255 TI - The dilemma of transvenous cardiac rhythm devices in hemodialysis patients: time to consider the epicardial approach? PMID- 21625257 TI - Strengths and weaknesses of renal markers as risk factors and surrogate markers. AB - The reports of associations between albuminuria and estimated glomerular filtration rate and mortality are a major advance in understanding the progression of renal disease and its sequelae. These findings reinforce the importance of population risk profiling. However, application is limited by validity concerns due to substantial unexplained heterogeneity in many meta analyses. Application to treatment decisions is premature given insufficient analysis of randomized trials of the association of by-arm differences in laboratory measures and by-arm differences in patient-relevant outcomes. PMID- 21625258 TI - PPAR-alpha transcriptional activity is required to combat doxorubicin-induced podocyte injury in mice. AB - Immunosuppressants and inhibitors of the renin angiotensin system are major reagents to treat nephrotic syndrome but their clinical effects are not necessarily satisfactory. Injection of doxorubicin in several strains of mice causes nephrotic syndrome-like disorder. Zhou et al. report that PPAR-alpha expression is downregulated in murine doxorubicin nephropathy and a PPAR-alpha agonist, fenofibrate, partially ameliorates the disorder induced likely through stabilization of nephrin expression and suppression of apoptosis in podocytes, providing a new preventive strategy. PMID- 21625260 TI - Sirolimus: not so sparing in the Spare-the-Nephron trial. PMID- 21625261 TI - Does renalase degrade catecholamines? PMID- 21625263 TI - Angiotensin II-induced phosphorylation of the sodium chloride cotransporter. PMID- 21625265 TI - A circuitous detour. PMID- 21625266 TI - The Case ? The smoker and the nephrologist. PMID- 21625267 TI - Inhibitory member of the apoptosis-stimulating protein of p53 (ASPP) family promotes growth and tumorigenesis in human p53-deficient prostate cancer cells. AB - iASPP is a member of the apoptosis-stimulating protein of p53 (ASPP) family and an evolutionarily conserved inhibitor of p53. Higher levels of iASPP proteins were examined in paraffin-embedded sections collected from 30 patients with prostate cancer using an immunohistochemical method. We found that specially knocking down iASPP with lentivirus-mediated small interfering RNA inhibited the growth, in vitro colony-forming capacity and in vivo tumorigenesis of p53 defective prostate cancer cells. Importantly, inhibition of iASPP induced cell apoptosis, which confers the inhibitory effect on cell survival. We conclude that iASPP is essential for prostate cancer cellular proliferation and survival and may be a potential target for the gene therapy for prostate cancer. PMID- 21625268 TI - Putting muscle in DNA methylation. PMID- 21625269 TI - The methyl-CpG-binding protein CIBZ suppresses myogenic differentiation by directly inhibiting myogenin expression. AB - Postnatal growth and regeneration of skeletal muscle are carried out mainly by satellite cells, which, upon stimulation, begin to express myogenin (Myog), the critical determinant of myogenic differentiation. DNA methylation status has been associated with the expression of Myog, but the causative mechanism remains almost unknown. Here, we report that the level of CIBZ, a methyl-CpG-binding protein, decreases upon myogenic differentiation of satellite-derived C2C12 cells, and during skeletal muscle regeneration in mice. We present data showing that the loss of CIBZ promotes myogenic differentiation, whereas exogenous expression of CIBZ impairs it, in cultured cells. CIBZ binds to a Myog promoter proximal region and inhibits Myog transcription in a methylation-dependent manner. These data suggest that the suppression of myogenic differentiation by CIBZ is dependent, at least in part, on the regulation of Myog. Our data show that the methylation status of this proximal Myog promoter inversely correlates with Myog transcription in cells and tissues, and during postnatal growth of skeletal muscle. Notably, induction of Myog transcription by CIBZ suppression is independent of the demethylation of CpG sites in the Myog promoter. These observations provide the first reported molecular mechanism illustrating how Myog transcription is coordinately regulated by a methyl-CpG-binding protein and the methylation status of the proximal Myog promoter. PMID- 21625270 TI - Selective serotonin reuptake inhibitors for the treatment of hypersensitive esophagus: a randomized, double-blind, placebo-controlled study. AB - OBJECTIVES: Ambulatory 24-h pH-impedance monitoring can be used to assess the relationship of persistent symptoms and reflux episodes, despite proton pump inhibitor (PPI) therapy. Using this technique, we aimed to identify patients with hypersensitive esophagus and evaluate the effect of selective serotonin reuptake inhibitors (SSRIs) on their symptoms. METHODS: Patients with normal endoscopy and typical reflux symptoms (heartburn, chest pain, and regurgitation), despite PPI therapy twice daily, underwent 24-h pH-impedance monitoring. Distal esophageal acid exposure (% time pH <4) was measured and reflux episodes were classified into acid or non-acid. A positive symptom index (SI) was declared if at least half of the symptom events were preceded by reflux episodes. Patients with a normal distal esophageal acid exposure time, but with a positive SI were classified as having hypersensitive esophagus and were randomized to receive citalopram 20 mg or placebo once daily for 6 months. RESULTS: A total of 252 patients (150 females (59.5%); mean age 55 (range 18-75) years) underwent 24-h pH impedance monitoring. Two hundred and nineteen patients (86.9%) recorded symptoms during the study day, while 105 (47.9%) of those had a positive SI (22 (20.95%) with acid, 5 (4.76%) with both acid and non-acid, and 78 (74.29%) with non-acid reflux). Among those 105 patients, 75 (71.4%) had normal distal esophageal acid exposure time and were randomized to receive citalopram 20 mg (group A, n=39) or placebo (group B, n=36). At the end of the follow-up period, 15 out of the 39 patients of group A (38.5%) and 24 out of the 36 patients of group B (66.7%) continue to report reflux symptoms (P=0.021). CONCLUSIONS: Treatment with SSRIs is effective in a select group of patients with hypersensitive esophagus. PMID- 21625271 TI - Effectiveness of combined pharmacologic and ligation therapy in high-risk patients with acute esophageal variceal bleeding. AB - OBJECTIVES: After an acute variceal bleeding, early decision for aggressive management of patients with worse prognosis may improve outcomes. The effectiveness of currently recommended standard therapy (drugs plus endoscopic ligation) for different risk subgroups and the validity of available risk criteria in clinical practice are unknown. METHODS: We analyzed data of 301 consecutive cirrhotic patients admitted with esophageal variceal bleeding. All patients received antibiotics, somatostatin, and in 263 early endoscopic therapy. A stratified 6-week mortality assessment according to risk (low-risk: Child-Pugh B without active bleeding or Child-Pugh A; high-risk: Child-Pugh B with active bleeding or Child-Pugh C) was performed. A multivariate analysis was conducted to elaborate a new risk classification rule. RESULTS: Among the 162 patients receiving emergency ligation, 14% rebled and 16% died. Standard therapy was very effective in all risk strata, even in high-risk patients, specially if eligible for therapeutic trials (child <14, age <=75 years, creatinine <=3.0 mg/dl, no hepatocellular carcinoma, or portal thrombosis), showing this stratum a 10% mortality. In patients receiving ligation, Child-Pugh C patients with baseline creatinine <1.0 mg/dl showed similar mortality to Child-Pugh A or B patients (8% vs. 7%, respectively). Only Child-Pugh C patients with creatinine >=1.0 were at a significant higher risk (Child-Pugh C: 46% mortality if creatinine >=1.0 vs. 8% if creatinine <1.0, P=0.006). CONCLUSIONS: The combination of somatostatin, antibiotics, and endoscopic ligation after an acute variceal bleeding in a real life situation is associated with very low mortality. Child-Pugh C patients with baseline creatinine >=1.0 mg/dl should be considered high-risk patients in this setting. PMID- 21625272 TI - Repercussions of early versus late initiation of dialysis. AB - Despite the widespread use of chronic dialysis, there remains a lack of consensus about the optimal time for initiation of renal replacement therapy. Recommendations from the National Kidney Foundation Kidney Disease Outcome Quality Initiative are generally used as the guideline. This has resulted in a trend in the past decade toward earlier initiation of dialysis, especially in the elderly. The associated burden of comorbidities in the elderly population has resulted in greatly reduced survival outcomes. Here, the data obtained from retrospective cohort studies have been corroborated with a recent randomized control trial conducted in Australia and New Zealand (IDEAL study). There are limitations to consider from the IDEAL study that originate from different confounding factors that intervene in the test population. The present paper is an evidence-based review of the literature, focusing on diminution of survival outcomes following the early initiation of dialysis. PMID- 21625273 TI - The birth of an open access multidisciplinary online journal. PMID- 21625274 TI - Provocative discography: Current status. AB - Low back pain is a common clinical problem that may be due to a variety of causes, including disc disease. Provocative discography is an imaging-guided procedure in which a contrast agent is injected into the nucleus pulposus of the disc. Despite its controversial history, it remains the only imaging technique that provides both anatomical and functional information about a diseased disc. Disc morphology is usually assessed on either radiographs or computed tomography (CT), or both. Functional evaluation of the disc consists of pain provocation and careful assessment of the patient's response to pain. As provocative discography is an invasive procedure, it should not be used as a screening study in patients with back pain. It should instead be reserved for carefully- selected patients whose painful symptoms cannot be explained by findings on non-invasive imaging modalities such as magnetic resonance imaging or CT, and who are not responsive to conservative measures. Discography is helpful in selection of patients and disc levels to be operated upon. Careful application of indications and meticulous technique are however required if a successful outcome is to be expected. PMID- 21625275 TI - Improved fracture detection using the mammographic film-screen combination. AB - AIM: The single emulsion or single screen system is usually reserved for mammography since its use in general radiography is limited. The purpose of this study is to compare the mammographic film-screen combination (MFC) and the standard film-screen combination (SFC) in terms of fracture and soft tissue injuries detection. PATIENTS, METHODS AND MATERIALS: In this prospective study, 41 patients from Accident and Emergency suspected of having injury in the hands, wrists, ankles and feet regions were radiographed using both MFC and SFC. These were compared in terms of image quality, presence of fractures and soft tissue injuries. The two different film-screen combinations were also compared in terms of detection of bony fragments, film characteristics such as film speed, contrast and spatial resolution, dose and cost. RESULTS: The MFC gives statistically better image quality compared to SFC. In 10% of patients, fractures were detected only in the MFC, which also detects tiny bone fragments that may not be resolved by the SFC. The spatial resolution of the MFC is greater than the SFC. The film speed and contrast of the MFC are lower than that of the SFC. The doses of MFC were higher compared to SFC. CONCLUSIONS: The MFC detects fractures better compared with SFC. However, the entrance skin dose for the mammographic film screen combination was about 35% to 55% higher than the standard film-screen combination. PMID- 21625276 TI - Phytobezoar: an unusual cause of intestinal obstruction. AB - Small bowel phytobezoars are rare and almost always obstructive. There have been previously reported cases of phytobezoars in the literature, however there are few reports on radiological findings for small bowel bezoars. Barium studies characteristically show an intraluminal filling defect of variable size that is not fixed to the bowel wall with barium filling the interstices giving a mottled appearance. On CT scan, the presence of a round or ovoid intraluminal mass with a 'mottled gas' pattern is believed to be pathognomonic. Since features on CT scans are characteristics and physical findings are of little assistance in the diagnosis of bezoar, the diagnostic value of CT needs to be emphasised. PMID- 21625277 TI - Intracystic papillary carcinoma of the breast. PMID- 21625278 TI - Malignant ovarian mixed germ cell tumour: a rare combination. AB - Ovarian germ cell tumours are very rare and affect mainly young girls and women. Due to this, the conservation of reproductive potential is of great concern. One of the most remarkable advances in oncology is in the treatment of malignant ovarian germ cell tumours. Two histological groups are distinguished: dygerminomas, equivalent to testicular seminomas, and non-dysgerminomatous tumours. We report a case of a 30-year-old nulliparous woman who presented with persistent per vaginal bleeding and was found to have a malignant mixed germ cell tumour comprising of both embryonal carcinoma and choriocarcinoma. PMID- 21625279 TI - Necrotising epididymo-orchitis with scrotal abscess. PMID- 21625280 TI - Practical guidelines for radiographers to improve computed radiography image quality. AB - Computed Radiography (CR) has become a major digital imaging modality in a modern radiological department. CR system changes workflow from the conventional way of using film/screen by employing photostimulable phosphor plate technology. This results in the changing perspectives of technical, artefacts and quality control issues in radiology departments. Guidelines for better image quality in digital medical enterprise include professional guidelines for users and the quality control programme specifically designed to serve the best quality of clinical images. Radiographers who understand technological shift of the CR from conventional method can employ optimization of CR images. Proper anatomic collimation and exposure techniques for each radiographic projection are crucial steps in producing quality digital images. Matching image processing with specific anatomy is also important factor that radiographers should realise. Successful shift from conventional to fully digitised radiology department requires skilful radiographers who utilise the technology and a successful quality control program from teamwork in the department. PMID- 21625281 TI - Sustaining and strengthening biij. PMID- 21625282 TI - In vivo molecular targeted radiotherapy. AB - Unsealed radionuclides have been in clinical therapeutic use for well over half a century. Following the early inappropriate clinical administrations of radium salts in the early 20th century, the first real clinical benefits became evident with the use of (131)I-sodium iodide for the treatment of hypothyroidism and differentiated thyroid carcinoma and (32)P-sodium phosphate for the treatment of polycythaemia vera. In recent years the use of bone seeking agents (89)Sr, (153)Sm and (186)Re for the palliation of bone pain have become widespread and considerable progress has been evident with the use of (131)I-MIBG and (90)Y somatostatin receptor binding agents. Although the use of monoclonal antibody based therapeutic products has been slow to evolve, the start of the 21st century has witnessed the first licensed therapeutic antibody conjugates based on (90)Y and (131)I for the treatment of non-Hodgkin's lymphoma. The future clinical utility of this form of therapy will depend upon the development of radiopharmaceutical conjugates capable of selective binding to molecular targets. The availability of some therapeutic radionuclides such as (188)Re produced from the tungsten generator system which can produce activity as required over many months, may make this type of therapy more widely available in some remote and developing countries.Future products will involve cytotoxic radionuclides with appropriate potency, but with physical characteristics that will enable the administration of therapeutic doses with the minimal need for patient isolation. Further developments are likely to involve molecular constructs such as aptamers arising from new developments in biotechnology.Patient trials are still underway and are now examining new methods of administration, dose fractionation and the clinical introduction of alpha emitting radiopharmaceutical conjugates. This review outlines the history, development and future potential of these forms of therapy. PMID- 21625283 TI - Epstein-Barr virus and Hodgkin's lymphoma in Cairo, Egypt. AB - Fifty-five consecutive cases of Hodgkin's lymphoma (HL), collected between 1996 and 1998 from Cairo, Egypt, were histologically subtyped, phenotyped, and then studied for the presence of Epstein-Barr virus (EBV). We used immunohistochemical stains for EBV latent membrane protein 1 (LMP-1) and in situ hybridization stains for EBV-encoded small RNA (EBER-1) transcripts. Forty-five cases (82%) had classic HL (cHL), and ten cases (18%) had nodular lymphocyte predominant HL (NLPHL), with each group expressing its typical phenotype. LMP-1 stains were positive in 63% and 0% of cHL and NLPHL cases, respectively. EBER-positive Reed Sternberg cells and variants were also present in 62% and 0% of each group, respectively. The cHL cases showed variable EBER positivity: nodular sclerosis, 58%; mixed cellularity, 100%; lymphocyte depletion, 100%; and unclassifiable, 67%. Our findings are similar to those from other developing countries and point towards a pathogenic role of EBV in cHL. PMID- 21625284 TI - Host Cell Preference of Toxoplasma gondii Cysts in Murine Brain: A Confocal Study. AB - Toxoplasma gondii is a protozoan parasite that is widely prevalent in humans and typically results in a chronic infection characterized by cysts located predominantly in the central nervous system. In immunosuppressed hosts, such as patients with HIV infection, the infection can be reactivated from the cysts in the brain resulting in a severe and potentially fatal encephalitis. Studies suggest that the chronic infection may also have neuropathological and behavioral effects in immune competent hosts. An improved understanding of tissue cyst behavior is of importance for understanding both the reactivation as well as the neurophysiological consequences of chronic infection. In vivo studies have identified neurons as host cells for cysts but in vitro studies have found that astrocytes can also foster development of the cysts. In this study we have addressed the question of which neural cell tissue cysts of T. gondii reside during chronic infection using a mouse model. Mice were infected with Me49 Strain T. gondii and the intracellular localization of the cysts analyzed during the development and establishment of a chronic infection at 1, 2, and 6 months post infection. Brains were fixed, cryosectioned, and stained with FITC-Dolichos biflorans to identify the Toxoplasma cysts and they were labeled with cell specific antibodies to neurons or astrocytes and then analyzed using confocal fluorescence microscopy. Cysts were found to occur almost exclusively in neurons throughout chronic infection. No cysts were identified in astrocytes, using the astrocyte marker, GFAP. Astrocyte interactions with neuronal-cysts, however, were frequently observed. PMID- 21625285 TI - Securely and Flexibly Sharing a Biomedical Data Management System. AB - Biomedical database systems need not only to address the issues of managing complex data, but also to provide data security and access control to the system. These include not only system level security, but also instance level access control such as access of documents, schemas, or aggregation of information. The latter is becoming more important as multiple users can share a single scientific data management system to conduct their research, while data have to be protected before they are published or IP-protected. This problem is challenging as users' needs for data security vary dramatically from one application to another, in terms of who to share with, what resources to be shared, and at what access level. We develop a comprehensive data access framework for a biomedical data management system SciPort. SciPort provides fine-grained multi-level space based access control of resources at not only object level (documents and schemas), but also space level (resources set aggregated in a hierarchy way). Furthermore, to simplify the management of users and privileges, customizable role-based user model is developed. The access control is implemented efficiently by integrating access privileges into the backend XML database, thus efficient queries are supported. The secure access approach we take makes it possible for multiple users to share the same biomedical data management system with flexible access management and high data security. PMID- 21625286 TI - The Impact of Job Stress on Smoking and Quitting: Evidence from the HRS. AB - Job-related stress might affect smoking behavior because smoking may relieve stress and stress can make individuals more present-focused. Alternatively, individuals may both self-select into stressful jobs and choose to smoke based on unobserved factors. We use data from the Health and Retirement Study to examine how job stress affects the probability that smokers quit and the number of cigarettes smoked for current smokers. To address the potential endogeneity of job stress based on time invariant factors, we include individual fixed effects, which control for factors such as ability to handle stress. Occupational fixed effects are also included to control for occupational characteristics other than stress; time dummies control for the secular decline in smoking rates. Using a sample of people who smoked in the previous wave, we find that job stress is positively related to continuing to smoke among recent smokers. The results indicate that the key impact of stress is on the extensive margin of smoking, as opposed to the number of cigarettes smoked. PMID- 21625287 TI - Continuous monitoring of changes in plasma nitrite following cerebral ischemia in a rabbit embolic stroke model. AB - In this proof-of-concept study, we investigated direct, continuous monitoring of plasma nitrite as an indicator of cerebral ischemia following clot embolization of rabbits via an indwelling carotid catheter. Two groups of rabbits were studied to compare the effects of embolization on nitrite levels. In the control group, blood was continuously obtained from a jugular venous catheter. The blood was immediately passed through an ultrafiltration filter; the filtrate was chemically reduced to convert free nitrite to nitric oxide (NO) and then measured using a NO specific electrode. In the embolized group, after a baseline nitrite level was achieved, blood clots were injected into the brain via the carotid artery catheter, and then nitrite levels were continuously measured from jugular venous blood. The stroke group showed a significantly greater increase in nitrite as compared to controls (p=0.017). Using the area-under-the-curve (AUC) method, results reached statistical significance (p<0.05) within 3 min of embolization. In embolized rabbits, NO(2) levels increased 424+/-256% compared to baseline. This study shows that nitrite can be measured immediately following a stroke and that our system measures nitrite independent of the extent of the stroke. This study provides evidence for the feasibility of using nitrite as a marker of ischemic stroke. PMID- 21625288 TI - High-throughput single-molecule fluorescence spectroscopy using parallel detection. AB - Solution-based single-molecule fluorescence spectroscopy is a powerful new experimental approach with applications in all fields of natural sciences. The basic concept of this technique is to excite and collect light from a very small volume (typically femtoliter) and work in a concentration regime resulting in rare burst-like events corresponding to the transit of a single-molecule. Those events are accumulated over time to achieve proper statistical accuracy. Therefore the advantage of extreme sensitivity is somewhat counterbalanced by a very long acquisition time. One way to speed up data acquisition is parallelization. Here we will discuss a general approach to address this issue, using a multispot excitation and detection geometry that can accommodate different types of novel highly-parallel detector arrays. We will illustrate the potential of this approach with fluorescence correlation spectroscopy (FCS) and single-molecule fluorescence measurements obtained with different novel multipixel single-photon counting detectors. PMID- 21625289 TI - A Graphical Model to Determine the Subcellular Protein Location in Artificial Tissues. AB - Location proteomics is concerned with the systematic analysis of the subcellular location of proteins. In order to perform comprehensive analysis of all protein location patterns, automated methods are needed. With the goal of extending automated subcellular location pattern analysis methods to high resolution images of tissues, 3D confocal microscope images of polarized CaCo2 cells immunostained for various proteins were collected. A three-color staining protocol was developed that permits parallel imaging of proteins of interest as well as DNA and the actin cytoskeleton. The collection is composed of 11 to 21 images for each of the 9 proteins that depict major subcellular patterns. A classifier was trained to recognize the subcellular location pattern of segmented cells with an accuracy of 89.2%. Using the Prior Updating method allowed improvement of this accuracy to 99.6%. This study demonstrates the benefit of using a graphical model approach for improving the pattern classification in tissue images. PMID- 21625291 TI - fgui: A Method for Automatically Creating Graphical User Interfaces for Command Line R Packages. AB - The fguiR package is designed for developers of R packages, to help rapidly, and sometimes fully automatically, create a graphical user interface for a command line R package. The interface is built upon the Tcl/Tk graphical interface included in R. The package further facilitates the developer by loading in the help files from the command line functions to provide context sensitive help to the user with no additional effort from the developer. Passing a function as the argument to the routines in the fgui package creates a graphical interface for the function, and further options are available to tweak this interface for those who want more flexibility. PMID- 21625292 TI - MDPs with Non-Deterministic Policies. AB - Markov Decision Processes (MDPs) have been extensively studied and used in the context of planning and decision-making, and many methods exist to find the optimal policy for problems modelled as MDPs. Although finding the optimal policy is sufficient in many domains, in certain applications such as decision support systems where the policy is executed by a human (rather than a machine), finding all possible near-optimal policies might be useful as it provides more flexibility to the person executing the policy. In this paper we introduce the new concept of non-deterministic MDP policies, and address the question of finding near-optimal non-deterministic policies. We propose two solutions to this problem, one based on a Mixed Integer Program and the other one based on a search algorithm. We include experimental results obtained from applying this framework to optimize treatment choices in the context of a medical decision support system. PMID- 21625293 TI - Image Atlas Construction via Intrinsic Averaging on the Manifold of Images. AB - In this paper, we propose a novel algorithm for computing an atlas from a collection of images. In the literature, atlases have almost always been computed as some types of means such as the straightforward Euclidean means or the more general Karcher means on Riemannian manifolds. In the context of images, the paper's main contribution is a geometric framework for computing image atlases through a two-step process: the localization of mean and the realization of it as an image. In the localization step, a few nearest neighbors of the mean among the input images are determined, and the realization step then proceeds to reconstruct the atlas image using these neighbors. Decoupling the localization step from the realization step provides the flexibility that allows us to formulate a general algorithm for computing image atlas. More specifically, we assume the input images belong to some smooth manifold M modulo image rotations. We use a graph structure to represent the manifold, and for the localization step, we formulate a convex optimization problem in R(k) (k the number of input images) to determine the crucial neighbors that are used in the realization step to form the atlas image. The algorithm is both unbiased and rotation-invariant. We have evaluated the algorithm using synthetic and real images. In particular, experimental results demonstrate that the atlases computed using the proposed algorithm preserve important image features and generally enjoy better image quality in comparison with atlases computed using existing methods. PMID- 21625294 TI - Physically-based Surface Texture Synthesis Using a Coupled Finite Element System. AB - This paper describes a stable and robust finite element solver for physically based texture synthesis over arbitrary manifold surfaces. Our approach solves the reaction-diffusion equation coupled with an anisotropic diffusion equation over surfaces, using a Galerkin based finite element method (FEM). This method avoids distortions and discontinuities often caused by traditional texture mapping techniques, especially for arbitrary manifold surfaces. Several varieties of textures are obtained by selecting different values of control parameters in the governing differential equations, and furthermore enhanced quality textures are generated by fairing out noise in input surface meshes. PMID- 21625290 TI - Mechanisms of Kaposi's Sarcoma-Associated Herpesvirus Latency and Reactivation. AB - The life cycle of Kaposi's sarcoma-associated herpesvirus (KSHV) consists of latent and lytic replication phases. During latent infection, only a limited number of KSHV genes are expressed. However, this phase of replication is essential for persistent infection, evasion of host immune response, and induction of KSHV-related malignancies. KSHV reactivation from latency produces a wide range of viral products and infectious virions. The resulting de novo infection and viral lytic products modulate diverse cellular pathways and stromal microenvironment, which promote the development of Kaposi's sarcoma (KS). The mechanisms controlling KSHV latency and reactivation are complex, involving both viral and host factors, and are modulated by diverse environmental factors. Here, we review the cellular and molecular basis of KSHV latency and reactivation with a focus on the most recent advancements in the field. PMID- 21625295 TI - A Regions of Confidence Based Approach to Enhance Segmentation with Shape Priors. AB - We propose an improved region based segmentation model with shape priors that uses labels of confidence/interest to exclude the influence of certain regions in the image that may not provide useful information for segmentation. These could be regions in the image which are expected to have weak, missing or corrupt edges or they could be regions in the image which the user is not interested in segmenting, but are part of the object being segmented. In the training datasets, along with the manual segmentations we also generate an auxiliary map indicating these regions of low confidence/interest. Since, all the training images are acquired under similar conditions, we can train our algorithm to estimate these regions as well. Based on this training we will generate a map which indicates the regions in the image that are likely to contain no useful information for segmentation. We then use a parametric model to represent the segmenting curve as a combination of shape priors obtained by representing the training data as a collection of signed distance functions. We evolve an objective energy functional to evolve the global parameters that are used to represent the curve. We vary the influence each pixel has on the evolution of these parameters based on the confidence/interest label. When we use these labels to indicate the regions with low confidence; the regions containing accurate edges will have a dominant role in the evolution of the curve and the segmentation in the low confidence regions will be approximated based on the training data. Since our model evolves global parameters, it improves the segmentation even in the regions with accurate edges. This is because we eliminate the influence of the low confidence regions which may mislead the final segmentation. Similarly when we use the labels to indicate the regions which are not of importance, we will get a better segmentation of the object in the regions we are interested in. PMID- 21625296 TI - Theoretical Analysis of Heuristic Search Methods for Online POMDPs. AB - Planning in partially observable environments remains a challenging problem, despite significant recent advances in offline approximation techniques. A few online methods have also been proposed recently, and proven to be remarkably scalable, but without the theoretical guarantees of their offline counterparts. Thus it seems natural to try to unify offline and online techniques, preserving the theoretical properties of the former, and exploiting the scalability of the latter. In this paper, we provide theoretical guarantees on an anytime algorithm for POMDPs which aims to reduce the error made by approximate offline value iteration algorithms through the use of an efficient online searching procedure. The algorithm uses search heuristics based on an error analysis of lookahead search, to guide the online search towards reachable beliefs with the most potential to reduce error. We provide a general theorem showing that these search heuristics are admissible, and lead to complete and epsilon-optimal algorithms. This is, to the best of our knowledge, the strongest theoretical result available for online POMDP solution methods. We also provide empirical evidence showing that our approach is also practical, and can find (provably) near-optimal solutions in reasonable time. PMID- 21625297 TI - A New Mechanism for Tracking Publicly Available Study Volunteer Demographics. AB - The importance of gathering and monitoring aggregate demographic data on the annual population of study volunteers in FDA-regulated clinical trials is widely acknowledged. To date, no formal mechanism exists to capture this information. The Tufts Center for the Study of Drug Development identified and tested a publicly available source of information on clinical trial participant data, NDA Reviews stored in the FDA's drugs@FDA database, to determine its accuracy, reliability, and feasibility. Thirty-seven new drug applications approved between 2006 and 2008 were evaluated and compared with published sources of demographic data. The authors conclude that the approach described here-NDA review extraction provides reasonably reliable and conservative estimates of study volunteer demographics and can serve as a useful baseline until Clinicaltrials.gov or other, more complete, public sources become available. PMID- 21625298 TI - Phasor-based single-molecule fluorescence lifetime imaging using a wide-field photon-counting detector. AB - Fluorescence lifetime imaging (FLIM) is a powerful approach to studying the immediate environment of molecules. For example, it is used in biology to study changes in the chemical environment, or to study binding processes, aggregation, and conformational changes by measuring Forster resonance energy transfer (FRET) between donor and acceptor fluorophores. FLIM can be acquired by time-domain measurements (time-correlated single-photon counting) or frequency-domain measurements (with PMT modulation or digital frequency domain acquisition) in a confocal setup, or with wide-field systems (using time-gated cameras). In the best cases, the resulting data is analyzed in terms of multicomponent fluorescence lifetime decays with demanding requirements in terms of signal level (and therefore limited frame rate). Recently, the phasor approach has been proposed as a powerful alternative for fluorescence lifetime analysis of FLIM, ensemble, and single-molecule experiments. Here we discuss the advantages of combining phasor analysis with a new type of FLIM acquisition hardware presented previously, consisting of a high temporal and spatial resolution wide-field single-photon counting device (the H33D detector). Experimental data with live cells and quantum dots will be presented as an illustration of this new approach. PMID- 21625299 TI - Hepatitis C-related arthropathy: Diagnostic and treatment considerations. AB - Hepatitis C-related arthropathy is one of the most common extrahepatic manifestations of hepatitis C virus (HCV) infection. Although symptoms can be disabling, the prognosis typically is benign. Patients who have atypical chronic inflammatory arthritis with an unknown cause should be evaluated for HCV infection. Testing for antibodies against cyclic citrullinated peptide is useful for distinguishing HCV-related arthropathy from rheumatoid arthritis. Early recognition of HCV infection greatly influences the selection of drug therapy. Although no clinical guidelines are available, many liver disease specialists favor using a stepwise approach to treatment. Future research efforts that focus on the pathogenesis of HCV-related arthropathy and novel therapeutic approaches are needed. (J Musculoskel Med. 2010;27:351-354). PMID- 21625300 TI - Perception of "Elliptical Speech" Following Cochlear Implantation: Use of Broad Phonetic Categories in Speech Perception. AB - This study investigated the perception of elliptical speech (Miller & Nicely, 1955) in an adult cochlear implant patient. A group of 20 adult listeners with normal hearing were used for comparison. Two experiments were conducted using sets of meaningful and anomalous English sentences. Two versions of each set of sentences were constructed: One set contained correct place of articulation cues; the other was transformed into elliptical speech using a procedure in which different places of articulation were all converted to alveolar place of articulation. The patient, "Mr. S," completed a same-different discrimination task and a sentence transcription task. The listeners with normal hearing completed both tasks under masking noise and low-pass filtering. In the same different task, under both conditions of signal degradation, listeners with normal hearing labeled a sentence with intact place of articulation cues and its elliptical version as the same. Mr. S also showed the same pattern. These findings support the claim by Miller and Nicely (1955) that under conditions of signal degradation, ellipsis can no longer be detected. In the sentence transcription task, however, subjects with normal hearing showed better transcription performance for sentences with intact place of articulation cues than for elliptical speech sentences, which was unexpected given the findings from the sentence discrimination experiment. Mr. S also showed the same pattern of performance. These new findings on the perception of elliptical speech suggest that cochlear implant users perceive speech and recognize spoken words using broad phonetic categories. PMID- 21625301 TI - Gender-Specific Barriers to Self-Sufficiency among Former Supplemental Security Income Drug Addiction and Alcoholism Beneficiaries: Implications for Welfare-To Work Programs and Services. AB - This study examines barriers to economic self-sufficiency among a panel of 219 former Supplemental Security Income (SSI) drug addiction and alcoholism (DA&A) recipients following elimination of DA&A as an eligibility category for SSI disability benefits. Study participants were comprehensively surveyed at six measurement points following the policy change. Generalized estimating equations were used to examine full-sample and gender-specific barriers to economic self sufficiency. Results indicate that access to transportation, age, and time are the strongest predictors of achieving self-sufficiency for both men and women leaving the welfare system. Gender-specific barriers are also identified. Future research needs to assess the generalizability of these results to other public assistance recipients. PMID- 21625302 TI - M=E and M=E Complexes of Iron and Cobalt that Emphasize Three-fold Symmetry (E = O, N, NR). AB - Mid-to-late transition metal complexes that feature terminal, multiply bonded ligands such as oxos, imides, and nitrides have been invoked as intermediates in several catalytic transformations of synthetic and biological significance. Until about ten years ago, isolable examples of such species were virtually unknown. Over the past decade or so, numerous chemically well-defined examples of such species have been discovered. In this context, the presentreview summarizes the development of 4- and 5-coordinate Fe(E) and Co(E) species under local three-fold symmetry. PMID- 21625303 TI - Spontaneous Pneumothorax in an Allogeneic Cell Transplant Recipient with Invasive Pulmonary Aspergillosis and Antecedent RSV Pneumonitis. AB - We report a case of invasive pulmonary aspergillosis (IPA) following respiratory syncytial virus infection in an allogeneic hematopoietic stem cell transplant (HSCT) recipient with chronic graft-versus-host disease. Delayed diagnosis of IPA resulted in the development of a pneumothorax, a rare consequence of fungal pneumonia. Respiratory virus infections are often harbingers of other infective organisms in HSCT recipients. More aggressive diagnostic investigations such as computed tomography scans of the thorax and bronchoscopy with bronchoalveolar lavage should be considered early in any HSCT patient presenting with respiratory virus pneumonia, particularly if atypical features are present or recovery is delayed. PMID- 21625304 TI - What are fungal infections? AB - Yeasts and moulds now rank amongst the 10 most frequently isolated pathogens in febrile patients with an impaired immune system. Fungi are mainly opportunistic pathogens that only invade the body if a severely weakened natural defense permits them to do so. Most factors facilitating an invasive fungal infection are unavoidable because they are directly connected to the underlying diseases as well as to their treatment.Modern aggressive treatment modalities jeopardize the defense mechanisms to an extent that even fungi with a low virulence may enter the body. PMID- 21625305 TI - Epidemiology of invasive fungal infections in the mediterranean area. AB - Although Candida species remain the relevant cause of IFI, other fungi (especially moulds) have become increasingly prevalent. In particular, Aspergillus species are the leading cause of mould infections but also Glomeromycota (formerly Zygomycetes) and Fusarium species are increasing in frequency, and are associated with high mortality rates. Many of these emerging infections occur as breakthrough infections in patients treated with new antifungal drugs. The causative pathogens, incidence rate and severity are dependent on the underlying condition, as well as on the geographic location of the patient population. France and Italy show the highest incident rates of Fusarium infections in Europe, following the US, where numbers are still increasing. Scedosporium prolificans, which primarily is found in soil in Spain and Australia, is most frequently isolated from blood cultures in a Spanish hospital. Geotrichum capitatum represents another species predominantly found in Europe with especially high rates in Mediterranean countries. The increasing resistance to antifungal drugs especially of these new emerging pathogens is a severe problem for managing these IFIs. PMID- 21625306 TI - Update on the laboratory diagnosis of invasive fungal infections. AB - Recent advances in the management of patients with haematological malignancies and transplant recipients have paralleled an increase in the incidence of fungal diseases due to pathogenic genera such as Candida and Aspergillus and the emergence of less common genera including Fusarium and Zygomycetes. Despite availability of new antifungal agents these opportunistic infections have high mortality. Rapid and reliable species identification is essential for antifungal treatment, but detection of the increasing diversity of fungal pathogens by conventional phenotypic methods remains difficult and time-consuming, and the results may sometimes be inconclusive, especially for unusual species. New diagnostic techniques (e.g., 1,3-beta-d-glucan detection) could improve this scenario, although further studies are necessary to confirm their usefulness in clinical practice. PMID- 21625307 TI - Fungal infections of the ear in immunocompromised host: a review. AB - Otomycosis is a fungal infection of the external ear; middle ear and open mastoid cavity.1 Meyer first described the fungal infection of the external ear canal in 1884. External ear canal has an ideal warm humid environment for the proliferation of fungus.2 Although this disease is rarely life threatening, it can presents a challenging and frustrating situation for the otologist and patients due to long term treatment and high rate of recurrence.3 Otomycosis is seen more frequently in immunocompromised patients as compared to immunocompetent persons. Recurrence rate is high in immunocompromised patients and they need longer duration treatment and complications are more frequent in immunocompromised patients.In the recent years; opportunistic fungal infections are gaining greater importance in human medicine as a result of possibly huge number of immunocompromised patients.4 In immunocompromised patients, it is important that the treatment of otomycosis be vigorous, to minimize complications such as hearing loss, tympanic membrane perforations and invasive temporal bone infection.5 Fungal cultures are essential to confirm the diagnosis.Hematological investigations play a very important role in confirming the diagnosis and immunity status of the patients. In diabetic patients with otomycosis, along with antifungal therapy, blood sugar levels should be controlled with medical therapy to prevent complications. PMID- 21625308 TI - Toxic epidermal necrolysis associated with severe cytomegalovirus infection in a patient on regular hemodialysis. AB - Primary illness with cytomegalovirus leads to latent infection with possible reactivations especially in the immunocompromised patients. Toxic epidermal necrolysis is an immune mediated cytotoxic reaction.A fifty years old female diabetic hypertensive patient with end stage renal disease was admitted with fever of unknown origin, constitutional symptoms, vague upper gastrointestinal symptoms and skin rash. Upper gastrointestinal endoscopic biopsy confirmed her diagnosis with cytomegalovirus esophagitis and duodenitis. Cytomegalovirus immunoglobulin M and immunoglobulin G levels were negative but polymerase chain reaction showed fulminant viremia. Biopsy of the skin rash was consistent with toxic epidermal necrolysis. Despite treatment with Ganciclovir, intravenous immunoglobulins, and granulocyte colony stimulating factor the patient's condition rapidly deteriorated and she died due to multiorgan failure, disseminated intravascular coagulopathy and overwhelming sepsis.Probably there is a true association linking toxic epidermal necrolysis to fulminant reactivation of cytomegalovirus. The aim of this anecdote is reporting a newly recognized presentation of cytomegalovirus. PMID- 21625309 TI - Too many mouldy joints - marijuana and chronic pulmonary aspergillosis. AB - Chronic pulmonary aspergillosis is a progressive debilitating disease with multiple underlying pulmonary diseases described. Here we report the association of chronic pulmonary aspergillosis and long term marijuana smoking in 2 patients and review the literature related to invasive and allergic aspergillosis. PMID- 21625310 TI - Brief Tale of a Bacteraemia by Rhodococcus equi, With Concomitant Lung Mass: What Came First, the Chicken or The Egg? AB - Rhodococcus equi is an uncommon Gram positive, variably acid-fast pathogen, that appears as hard to treat mostly owing to the establishment of intracellular niches. Lack of interpretive criteria for susceptibility testing may lead to under-reporting or overestimation of resistances, whereas knowledge about this pathogen's clinical impact may be affected by erroneous phenotype-based characterization at a genus and species level.We present the case of a bacteraemia with a concomitant lung mass in a lymphoma patient, that further highlights the emergence of rhodococcal diseases as a matter for concern in the fields of infectious diseases and haematology. PMID- 21625311 TI - Invasive candidiasis in non-hematological patients. AB - Candida is one of the most frequent pathogens isolated in bloodstream infections, and is associated with significant morbidity and mortality. In addition to haematological patients, there are several other populations with a substantial risk of developing invasive candidiasis (IC). These include patients undergoing prolonged hospitalisation with the use of broad-spectrum antibiotics, those fitted with intravascular catheters, admitted to both adult and neonate intensive care units (ICU) or gastrointestinal surgery wards and subjects with solid tumours undergoing cytotoxic chemotherapy. As a general rule, every immunocompromised patient might be at risk of Candida infection, including, for example, diabetic patients.The epidemiology of species responsible for IC has been changing, both at local and worldwide level, shifting from C. albicans to non-albicans species, that can be intrinsically resistant to fluconazole (C. krusei and, to some extent, C. glabrata), difficult to eradicate because of biofilm production (C. parapsilosis) or than might acquire resistance to azole during therapy.Delaying the specific therapy has been shown to increase morbidity and mortality, but traditional microbiological diagnosis is poorly sensitive and slow. Thus, culture-based treatment may result in therapy started too late. In order to reduce the mortality in IC, several management strategies have been developed: prophylaxis, empirical and pre-emptive therapy. Compared to prophylaxis, the latter approaches allow to reduce the use of antifungals by targeting only patients at very high risk of IC. Non-invasive serological markers and scores based on clinical prediction rules such as the presence of risk factors or Candida colonisation, have been developed with the aim of allowing prompt initiation of treatment. Although the use of these diagnostic tools in pre emptive strategies is promising, the performance and cost-effectiveness should be tested in large trials.Agents recommended for initial treatment of candidemia in severely ill patients include echinocandins and lipid formulations of amphotericin B, while stable patients without risk factors for azole-resistance might be treated with fluconazole. PMID- 21625312 TI - Therapy Outcome of a T-Cell-Rich-B-Cell Lymphoma (TCRBCL) Patient with R-CHOP in Ibadan, Nigeria: a Case Report. AB - T-cell-rich B-cell lymphoma (TCRBCL) is considered a rare variant of aggressive B cell lymphoma characterized by few neoplastic B cells and a large reactive infiltrate with striking similarities to nodular lymphocyte predominant Hodgkin's lymphoma.A case of a 46 year old man referred with a 5 months history of generalized lymphadenopathy, weight loss, low grade pyrexia and two separately reported lymph node histology consistent with TCRBCL is described.The clinical course was indeed aggressive because in spite of initial treatment with four cycles of CHOP combination chemotherapy, followed by R+CHOP(x 6 cycles), signs of tumor re-growth/infiltration were frequently observed. Also, recurrent infection was frequent, troublesome and eventually became overwhelming resulting to the loss of the patient.This case, being the first case of TCRBCL diagnosed by immunohistochemical confirmation and managed at this centre with R-CHOP, is presented to highlight the dilemma in making diagnosis, clinical challenges faced and rituximab therapy outcome especially in resource poor country. It will also serve to increase our index of suspicion and the need reinforce immunohistochemistry in the diagnosis of lymphoma. PMID- 21625313 TI - Severe eosinophilia in a case of giardiasis. PMID- 21625314 TI - Severe acute hepatitis B treated with entecavir. AB - Hepatitis B virus (HBV) infection constitutes a serious global health problem. Nowadays there are divergent data regarding the use of antiviral drugs to treat acute hepatitis B. We present here a case of a 62-year-old man affected by severe acute hepatitis B with progressive worsening of clinical and hepatic function. The patient was treated with entecavir without critical side effects. We observed rapid clinical and laboratory improvements and the disappearance of hepatitis B surface antigen (HBsAg). The treatment with entecavir was protracted until 17(th) week when the antibody anti-HBs appeared. Entecavir should be carefully considered for the treatment of severe acute hepatitis B cases. PMID- 21625315 TI - 5-azacytidine in chronic myelomonocytic leukemia: case report and review of literature. AB - Hypomethylating drugs are useful in the management of Myelodysplastic syndromes, but there are only few reports on chronic myelomonocycitic (CMML) leukemia patients. We describe our experience in 3 CMML patients treated with azacitidine. Two patients obtained partial response after 4 treatment cycles with only minor toxicity and are in continuous partial response, with stable peripheral blood counts, at 29 and 30 cycles from treatment start. PMID- 21625317 TI - Invasive Candida infections in patients with haematological malignancies and hematopoietic stem cell transplant recipients: current epidemiology and therapeutic options. AB - In the last decades, the global epidemiological impact of invasive candidiasis (IC) in patients with hematologic malignancies (HM) and in hematopoietic stem cell transplant (HSCT) recipients has decreased and the incidence of invasive aspergillosis exceeded that of Candida infections. The use of prevention strategies, first of all antifungal prophylaxis with triazoles, contributed to the reduction of IC in these populations as demonstrated by several epidemiological studies. However, relatively little is known about the current epidemiological patterns of IC in HM and HSCT populations, because recent epidemiological data almost exclusively derive from retrospective experiences and few prospective data are available. Several prospective, controlled studies in the prophylaxis of invasive fungal diseases have been conducted in both the HM and HSCT setting. On the contrary, most of the prospective controlled trials that demonstrated the efficacy of the antifungal drugs echinocandins and voriconazole in the treatment of candidemia and invasive candidiasis mainly involved patients with underlying conditions other than HM or HSCT. For these reasons, international guidelines provided specific indications for the prophylaxis strategies in HM and HSCT patients, whereas the recommendations on therapy of documented Candida infections are based on the results observed in the general population and should be considered with caution. PMID- 21625318 TI - Common parasite with uncommon associations. PMID- 21625316 TI - Zygomycosis in Immunocompromised non-Haematological Patients. AB - Zygomycoses caused by fungi of the mucorales order (mucormycoses) are emerging fungal diseases with a high fatality rate. The most important risk factors include neutropenia or functional neutropenia, diabetic ketoacidosis, iron overload, major trauma, prolonged use of corticosteroids, illicit intravenous drug (ID) use, neonatal prematurity, malnourishment, and maybe a previous exposure to antifungal agents with no activity against zygomycetes, such as voriconazole and echinocandins.A high index of suspicion is crucial for the diagnosis, as prompt and appropriate management can considerably reduce morbidity and mortality. Suspicion index can be increased through recognition of the differential patterns of clinical presentation. In the non- haematological immunocompromised patients, mucormycosis can manifest in various clinical forms, depending on the underlying condition: mostly as rhino-orbital or rhino-cerebral in diabetes patients, pulmonary infection in patients with malignancy or solid organ transplantation, disseminated infection in iron overloaded or deferoxamine treated patients, cerebral - with no sinus involvement - in ID users, gastrointestinal in premature infants or malnourishment, and cutaneous after direct inoculation in immunocompetent individuals with trauma or burns.Treating a patient's underlying medical condition and reducing immunosuppression are essential to therapy. Rapid correction of metabolic abnormalities is mandatory in cases such as uncontrolled diabetes, and corticosteroids or other immunosuppressive drugs should be discontinued where feasible. AmphotericinB or its newer and less toxic lipid formulations are the drugs of choice regarding antifungal chemotherapy, while extensive surgical debridement is essential to reduce infected and necrotic tissue. A high number of cases could be prevented through measures including diabetes control programmes and proper pre- and post surgical hygiene. PMID- 21625319 TI - Blood Vessel Bank: Organization and Function. PMID- 21625320 TI - Computer-Aided Grading of Lymphangioleiomyomatosis (LAM) using HRCT. AB - Lymphangioleiomyomatosis (LAM) is a multisystem disorder associated with proliferation of smooth muscle-like cells, which leads to destruction of lung parenchyma. Subjective grading of LAM on HRCT is imprecise and can be arduous especially in cases with severe involvement. We propose a computer-aided evaluation system that grades LAM involvement based on analysis of lung texture patterns. A committee of support vector machines is employed for classification. The system was tested on 36 patients. The computer grade demonstrates good correlation with subjective radiologist grade (R=0.91, p<0.0001) and pulmonary functional tests (R=0.85, p<0.0001). The grade also provides precise progression assessment of disease over time. PMID- 21625321 TI - Robust Biological Parametric Mapping: An Improved Technique for Multimodal Brain Image Analysis. AB - Mapping the quantitative relationship between structure and function in the human brain is an important and challenging problem. Numerous volumetric, surface, region of interest and voxelwise image processing techniques have been developed to statistically assess potential correlations between imaging and non-imaging metrics. Recently, biological parametric mapping has extended the widely popular statistical parametric approach to enable application of the general linear model to multiple image modalities (both for regressors and regressands) along with scalar valued observations. This approach offers great promise for direct, voxelwise assessment of structural and functional relationships with multiple imaging modalities. However, as presented, the biological parametric mapping approach is not robust to outliers and may lead to invalid inferences (e.g., artifactual low p-values) due to slight mis-registration or variation in anatomy between subjects. To enable widespread application of this approach, we introduce robust regression and robust inference in the neuroimaging context of application of the general linear model. Through simulation and empirical studies, we demonstrate that our robust approach reduces sensitivity to outliers without substantial degradation in power. The robust approach and associated software package provides a reliable way to quantitatively assess voxelwise correlations between structural and functional neuroimaging modalities. PMID- 21625322 TI - ? PMID- 21625323 TI - New insights into the prevention of staphylococcal infections and toxic shock syndrome. AB - Staphylococcus aureus is a major human pathogen capable of causing various diseases, from skin infections to life-threatening pneumonia and toxic shock syndrome. S. aureus exoproteins, including superantigens, contribute significantly to the pathogenesis of this organism. Antibiotics inhibit growth, but often provide no protection from S. aureus exoproteins. With the emergence of antibiotic-resistant S. aureus, new therapeutic options to treat or prevent S. aureus-associated diseases are critical. Most S. aureus infections begin on the skin or mucosal surfaces from direct inflammatory or cytotoxic effects of exotoxins. Therefore, antitoxin therapies that prevent toxin production and prevent their effects on host cells are being researched. Current treatments for staphylococcal diseases and recent developments in antitoxin therapeutic agents and vaccines are reviewed. PMID- 21625324 TI - Role of abnormal sarcoplasmic reticulum function in atrial fibrillation. AB - Atrial fibrillation (AF) is the most common cardiac arrhythmia, and is a cause of significant morbidity and mortality if left untreated. AF has been associated with profound changes in sarcoplasmic reticulum Ca(2+) homeostasis, which might contribute to both reduced contractile function and increased arrhythmogenesis in atria. Studies in human tissue samples and various animal models of AF have revealed changes in both expression levels and posttranslational modifications of key Ca(2+) handling proteins, which may contribute to arrhythmogenesis. In this review, we will focus on the molecular basis of alterations in sarcoplasmic reticulum Ca(2+) handling in AF and their potential therapeutic implications. PMID- 21625325 TI - Psychiatric molecular genetics and the ethics of social promises. AB - A recent literature review of commentaries and 'state of the art' articles from researchers in psychiatric genetics (PMG) offers a consensus about progress in the science of genetics, disappointments in the discovery of new and effective treatments, and a general optimism about the future of the field. I argue that optimism for the field of psychiatric molecular genetics (PMG) is overwrought, and consider progress in the field in reference to a sample estimate of US National Institute of Mental Health funding for this paradigm for the years 2008 and 2009. I conclude that the amounts of financial investment in PMG is questionable from an ethical perspective, given other research and clinical needs in the USA. PMID- 21625326 TI - TELEMEDICINA: UN DESAFIO PARA AMERICA LATINA. AB - La telemedicina es una tendencia creciente en la prestacion de los servicios medicos. Aunque la eficacia de esta practica no ha estado bien establecida, es probable que los paises en desarrollo compartiran este nuevo paradigma con los desarrollados. Los defensores de la telemedicina en America Latina sostienen que sera una herramienta util para reducir las disparidades y mejorar la accesibilidad de atencion de salud. Aunque America Latina quiza se convierta en un lugar para la investigacion e investigacion de estos procedimientos, no esta claro como la telemedicina podria contribuir a mejorar la accesibilidad para las poblaciones desfavorecidas, o coexistir con sistemas de atencion de salud publicos cronicamente enfermos.Telemedicine is a growing trend in the provision of medical services. Although the effectiveness of this practice has not been well established, it is likely that developing countries will share this new paradigm with developed ones. Supporters of telemedicine in Latin America maintain that it will be a useful tool for reducing disparities and improving health care accessibility. Although Latin America might become a place for research and investigation of these procedures, it is not clear how telemedicine could contribute to improving accessibility for disadvantaged populations, or coexist with chronically ill-funded public healthcare systems. PMID- 21625327 TI - Tripodal Binding Units for Self-Assembled Monolayers on Gold: A Comparison of Thiol and Thioether Headgroups. AB - Whereas thiols and thioethers are frequently used as binding units of oligodentate precursor molecules to fabricate self-assembled monolayers (SAMs) on coinage metal and semiconductor surfaces, their use for tridentate bonding configuration is still questionable. Against this background, novel tridentate thiol ligands, PhSi(CH(2)SH)(3) (PTT) and p-Ph-C(6)H(4)Si(CH(2)SH)(3) (BPTT), were synthesized and used as tripodal adsorbate molecules for the fabrication of SAMs on Au(111). These SAMs were characterized by X-ray photoelectron spectroscopy (XPS) and near edge X-ray absorption fine structure (NEXAFS) spectroscopy. The PTT and BPTT films were compared with the analogous systems comprised of same tripodal ligands with thioether instead of thiol binding units (anchors). XPS and NEXAFS data suggest that the binding uniformity, packing density, and molecular alignment of the thiol-based ligands in the respective SAMs is superior to their thioether counterparts. In addition, the thiol-based films showed significantly lower levels of contamination. Significantly, the quality of the PTT SAMs on Au(111) was found to be even higher than that of the films formed from the respective monodentate counterpart, benzenethiol. The results obtained allow for making some general conclusions on the specific character of molecular self-assembly in the case of tridentate ligands. PMID- 21625329 TI - Stability and Morphology of Gold Nanoisland Arrays Generated from Layer-by-Layer Assembled Nanoparticle Multilayer Films: Effects of Heating Temperature and Particle Size. AB - This article reports the effects of heating temperature and composition of nanoparticle multilayer films on the morphology, stability, and optical property of gold nanoisland films prepared by nanoparticle self-assembly/heating method. First, nanoparticle-polymer multilayer films are prepared by the layer-by-layer assembly. Nanoparticle multilayer films are then heated at temperature ranging from 500 degrees C to 625 degrees C in air to induce an evaporation of organic matters from the films. During the heating process, the nanoparticles on the solid surface undergo coalescence, resulting in the formation of nanostructured gold island arrays. Characterization of nanoisland films using atomic force microscopy and UV-vis spectroscopy suggests that the morphology and stability of gold island films change when different heating temperatures are applied. Stable gold nanoisland thin film arrays can only be obtained after heat treatments at or above 575 degrees C. In addition, the results show that the use of nanoparticles with different sizes produces nanoisland films with different morphologies. Multilayer films containing smaller gold nanoparticles tend to produce more monodisperse and smaller island nanostructures. Other variables such as capping ligands around nanoparticles and molecular weight of polymer linkers are found to have only minimal effects on the structure of island films. The adsorption of streptavidin on the biotin-functionalized nanoisland films is studied for examining the biosensing capability of nanoisland arrays. PMID- 21625328 TI - Etiology, Treatment and Prevention of Obesity in Childhood and Adolescence: A Decade in Review. AB - Childhood obesity has become an epidemic on a worldwide scale. This article gives an overview of the progress made in childhood and adolescent obesity research in the last decade, with a particular emphasis on the transdisciplinary and complex nature of the problem. The following topics are addressed: 1) current definitions of childhood and adolescent overweight and obesity; 2) demography of childhood and adolescent obesity both in the US and globally; 3) current topics in the physiology of fat and obesity; 4) psychosocial correlates of childhood and adolescent overweight and obesity; 5) the three major obesity-related behaviors, i.e. dietary intake, physical activity and sleep; 6) genes components of childhood and adolescent obesity; 7) environment and childhood and adolescent obesity; and 8) progress in interventions to prevent and treat childhood obesity. The article concludes with recommendations for future research, including the need for large-scale, high dose and long-term interventions that take into account the complex nature of the problem. PMID- 21625330 TI - ESTIMATION AND TESTING FOR PARTIALLY LINEAR SINGLE-INDEX MODELS. AB - In partially linear single-index models, we obtain the semiparametrically efficient profile least-squares estimators of regression coefficients. We also employ the smoothly clipped absolute deviation penalty (SCAD) approach to simultaneously select variables and estimate regression coefficients. We show that the resulting SCAD estimators are consistent and possess the oracle property. Subsequently, we demonstrate that a proposed tuning parameter selector, BIC, identifies the true model consistently. Finally, we develop a linear hypothesis test for the parametric coefficients and a goodness-of-fit test for the nonparametric component, respectively. Monte Carlo studies are also presented. PMID- 21625332 TI - Stability of Sodium Nitroprusside and Sodium Thiosulfate 1:10 Intravenous Admixture. AB - PURPOSE: Thiosulfate has been shown to reduce the risk of cyanide toxicity during nitroprusside administration. Admixtures containing both agents may provide a safe and effective alternative to more expensive agents used to reduce blood pressure in the critically ill patient. This study determined the physical and chemical stability of a 1:10 nitroprusside:thiosulfate admixture, stored up to 48 hours. The economic consequences of a shift toward using thiosulfate and nitroprusside, and away from higher cost alternatives, are considered. METHODS: Seven samples of 50 mg nitroprusside and 500 mg thiosulfate were prepared and stored away from light, at room temperature, and in a refrigerator prepared in D5W and NS. Each sample was analyzed via a novel high-performance liquid chromatographic (HPLC) method at time 0, 8, 24, and 48 hours. The method was tested and passed specifications for linearity, reproducibility, and accuracy. A visual inspection by 9 licensed pharmacists was used to demonstrate physical stability. A cost evaluation comparing nitroprusside and thiosulfate to alternative agents was completed. RESULTS: The concentration of both nitroprusside and thiosulfate remain greater than 95% of the initial concentration through 48 hours. Physical compatibility was confirmed in all samples tested through 72 hours. CONCLUSION: The combination of nitroprusside and thiosulfate is chemically and physically stable as a single compounded dose for up to 48 hours when stored at room temperature and protected from light. The admixture represents an inexpensive option to other higher cost alternatives such as nicardipine or clevidipine. PMID- 21625331 TI - Expanding the Antimalarial Drug Arsenal-Now, But How? AB - The number of available and effective antimalarial drugs is quickly dwindling. This is mainly because a number of drug resistance-associated mutations in malaria parasite genes, such as crt, mdr1, dhfr/dhps, and others, have led to widespread resistance to all known classes of antimalarial compounds. Unfortunately, malaria parasites have started to exhibit some level of resistance in Southeast Asia even to the most recently introduced class of drugs, artemisinins. While there is much need, the antimalarial drug development pipeline remains woefully thin, with little chemical diversity, and there is currently no alternative to the precious artemisinins. It is difficult to predict where the next generation of antimalarial drugs will come from; however, there are six major approaches: (i) re-optimizing the use of existing antimalarials by either replacement/rotation or combination approach; (ii) repurposing drugs that are currently used to treat other infections or diseases; (iii) chemically modifying existing antimalarial compounds; (iv) exploring natural sources; (v) large-scale screening of diverse chemical libraries; and (vi) through parasite genome-based ("targeted") discoveries. When any newly discovered effective antimalarial treatment is used by the populus, we must maintain constant vigilance for both parasite-specific and human-related factors that are likely to hamper its success. This article is neither comprehensive nor conclusive. Our purpose is to provide an overview of antimalarial drug resistance, associated parasite genetic factors (1. Introduction; 2. Emergence of artemisinin resistance in P. falciparum), and the antimalarial drug development pipeline (3. Overview of the global pipeline of antimalarial drugs), and highlight some examples of the aforementioned approaches to future antimalarial treatment. These approaches can be categorized into "short term" (4. Feasible options for now) and "long term" (5. Next generation of antimalarial treatment-Approaches and candidates). However, these two categories are interrelated, and the approaches in both should be implemented in parallel with focus on developing a successful, long-lasting antimalarial chemotherapy. PMID- 21625333 TI - The modified-bead stretched sample method: development and application to MALDI MS imaging of protein localization in the spinal cord. AB - Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) has been used to create spatial distribution maps from lipids, peptides, and proteins in a variety of biological tissues. MALDI-MSI often involves trade-offs between the extent of analyte extraction and desired spatial resolution, compromises that can adversely affect detectability. For example, increasing the extraction time can lead to unwanted analyte spatial redistribution. With the stretched sample method (SSM), the extraction period can be extended, resulting in reduced analyte redistribution while suppressing detection of cationic salt adducts. The SSM involves thaw-mounting a thin tissue section onto a substrate of small glass beads embedded in Parafilm M and then stretching the membrane to fragment the tissue into thousands of bead-sized pieces. Here, we applied the SSM method to MALDI-MSI using rat spinal cord as a model. We used surface-modified beads coated with trypsin or chymotrypsin in order to facilitate controlled digestion and detection of proteins. The enzymatic reactions were maintained by repeatedly condensing water on the stretched sample surface. As a result, new peptides formed by tryptic or chymotryptic protein digestion were detected and identified using a combination of MALDI-MSI and offline liquid chromatography tandem mass spectrometric analysis. Localization of these peptides indicated the distribution of their proteins of origin, including myelin basic protein, actin beta, and tubulin alpha chain. Additionally, we used uncoated beads to create distribution maps of many endogenous lipids and small peptides. The extension of the SSM using modified beads resulted in the creation of mosaic bead surfaces where adjacent beads were coated with different enzymes or other reactive chemicals, permitting investigation of the distributions of a wider range of analytes in biological samples within a single experiment. PMID- 21625334 TI - "To Numb Out and Start to Feel Nothing": Experiences of Stress Among Crack Cocaine Using Women in a Midwestern City. AB - The study uses qualitative interviews conducted with 19 crack using women to explore their experiences of stress and their views regarding the relationship between stress and drug use. Fifteen of the women participated in follow-up interviews conducted 5-7 years after the baseline. Life history interviews unveiled a pattern of close connection between the intensity of women's drug use and the level of stress they experienced in relation to their past adversities and current life circumstances. The majority of the women viewed stress as an important causal explanation of their drug use. Tensions related to romantic relationships, traumatic childhood, motherhood failures, unabated grief, and humiliating experiences of "crack life" were discussed as the most common sources of psychosocial stress. Most women had very limited positive coping resources and skills. Crack use was perceived as a very common, although highly maladaptive, way to deal with stress. Implications for interventions are discussed. PMID- 21625335 TI - Sleep-active cells in the cerebral cortex and their role in slow-wave activity. AB - We recently identified neurons in the cerebral cortex that become activated during sleep episodes with high slow-wave activity (SWA). The distinctive properties of these neurons are the ability to produce nitric oxide and their long-range projections within the cortex. In this review, we discuss how these characteristics of sleep-active cells could be relevant to SWA production in the cortex. We also discuss possible models of the role of nNOS cells in SWA production. PMID- 21625336 TI - Difference in Energy Metabolism of Annulus Fibrosus and Nucleus Pulposus Cells of the Intervertebral Disc. AB - Low back pain is associated with intervertebral disc degeneration. One of the main signs of degeneration is the inability to maintain extracellular matrix integrity. Extracellular matrix synthesis is closely related to production of adenosine triphosphate (i.e. energy) of the cells. The intervertebral disc is composed of two major anatomical regions: annulus fibrosus and nucleus pulposus, which are structurally and compositionally different, indicating that their cellular metabolisms may also be distinct. The objective of this study was to investigate energy metabolism of annulus fibrosus and nucleus pulposus cells with and without dynamic compression, and examine differences between the two cell types. Porcine annulus and nucleus tissues were harvested and enzymatically digested. Cells were isolated and embedded into agarose constructs. Dynamically loaded samples were subjected to a sinusoidal displacement at 2 Hz and 15% strain for 4 h. Energy metabolism of cells was analyzed by measuring adenosine triphosphate content and release, glucose consumption, and lactate/nitric oxide production. A comparison of those measurements between annulus and nucleus cells was conducted. Annulus and nucleus cells exhibited different metabolic pathways. Nucleus cells had higher adenosine triphosphate content with and without dynamic loading, while annulus cells had higher lactate production and glucose consumption. Compression increased adenosine triphosphate release from both cell types and increased energy production of annulus cells. Dynamic loading affected energy metabolism of intervertebral disc cells, with the effect being greater in annulus cells. PMID- 21625337 TI - Rab small GTPase emerges as a regulator of TOR complex 2. AB - In diverse eukaryotic species from yeast to human, TOR (Target Of Rapamycin) protein kinase operates in signaling pathways that link extracellular stimuli to the control of cell growth and metabolism. TOR kinase functions in two distinct protein complexes, TOR complex 1 (TORC1) and 2 (TORC2). While TORC1 is known to be under the control of the Ras-like small GTPase Rheb, our knowledge about TORC2 regulation is very limited. We thus set out to identify TORC2 activators through genetic approaches in the fission yeast Schizosaccharomyces pombe. Here we briefly review our study that has identified a Rab-family GTPase, Ryh1 and its GEF (guanine nucleotide exchange factor) as positive regulators of TORC2 signaling in S. pombe. Considering the evolutionary conservation of the TOR pathways, it is conceivable that Rabfamily GTPases also play a role in the regulation of human TORC2 in cellular proliferation and insulin signaling. PMID- 21625339 TI - "I No Like Get Caught Using Drugs": Explanations for Refusal as a Drug Resistance Strategy for Rural Native Hawaiian Youth. AB - This exploratory study examined the use of explanations for refusal as a drug resistance strategy for rural Native Hawaiian youth. Fourteen gender-specific focus groups were conducted within seven middle or intermediate schools on the Island of Hawai'i (N = 64). Participants were asked to describe drug resistance strategies in response to 15 hypothetical culturally specific drug-related problem scenarios developed from earlier research. The findings indicate that variations in the types of explanations used for refusal were based on the type of drug offerer in the associated scenario (i.e., peer/friend, cousin, or parent). Participants also described the rationales for the use of different explanations with different drug offerers. The findings suggest that culturally grounded drug prevention programs for Hawaiian youth should incorporate the use of specific types of explanations for refusal, depending on the youths' relationship to the drug offerer. PMID- 21625338 TI - An Update on the Biology of RAS/RAF Mutations in Colorectal Cancer. AB - Deaths caused by colorectal cancer (CRC) are among the leading causes of cancer related death in the United States and around the world. Approximately 150,000 Americans are diagnosed with CRC each year and around 50,000 will die from it. Mutations in many key genes have been identified that are important to the pathogenesis of CRC. Among the genes mutated in CRC, RAS and RAF mutations are common events. Both RAS and RAF are critical mediators of the mitogen-activated protein kinase (MAPK) pathway that is involved in regulating cellular homeostasis, including proliferation, survival, and differentiation. In this review, we provide a historical perspective and update on RAS/RAF mutations as related to colorectal cancer. Additionally, we will review recent mouse models of RAS and RAF mutations that have an impact on CRC research. PMID- 21625340 TI - A Comparative Assessment of Zootherapeutic Remedies From Selected Areas in Albania, Italy, Spain and Nepal. AB - Zootherapy is the treatment of human ailments with remedies derived from animals and their products. Despite its prevalence in traditional medical practices worldwide, research on this phenomenon has often been neglected in comparison to medicinal plant research. Interviews regarding zootherapeutic traditions were conducted with informants from Albania, Italy, Nepal and Spain. We identified 80 species used in zootherapeutic remedies, representing 4 phyla in the animal kingdom: Annelida, Arthropoda, Chordata, and Mollusca. Remedies were ranked by consensus indices. Our studies show that the selection of medicinal fauna is mediated by human subsistence patterns. Concepts of health and disease differ among our study sites in the Mediterranean and Asia, and these differences also play a substantive role in the selection and use of animal-based remedies. PMID- 21625341 TI - Maxillary sinus floor augmentation on humans: Packing simulations and 8 months histomorphometric comparative study of anorganic bone matrix and beta-tricalcium phosphate particles as grafting materials. AB - The present study compares the behaviour of an anorganic bone matrix material and a synthetic beta-Tricalcium phosphate employed as grafting materials in a sinus floor augmentation two step protocol in humans. In order to estimate the initial occupation level for the two materials, an 'in vitro' simulation has been performed to analyse macroporosity created due to particle packing in terms of porosity and interparticle distances. Grafting in the sinus floor augmentation was performed by filling the defects only with pure grafting materials without autogenous bone addition. The new-bone generated is 100% based on the osteoconductive properties of the grafted materials in contact with physiological fluids. The implants were placed 8 months after the grafting procedure. All the implanted positions were biopsied and embedded in methacrylate resin. Histomorphometric analyses were done over thin film undecalcified sections. Packing simulations allow establishing a comparison of the resorbed volumes related to the initial occupancy of the grafting materials inside the defect. The nature of this interconnected pore network is very alike for either material so new-bone generated was similar (~35 vol.%). PMID- 21625342 TI - Hospital Financial Conditions and the Provision of Unprofitable Services. AB - Increases in hospital financial pressure resulting from public and private payment policy may substantially reduce a hospital's ability to provide certain services that are not well compensated or are frequently used by the uninsured. The objective of this study is to examine the impact of hospital financial condition on the provision of these unprofitable services for the insured and uninsured. Economic theory provides the conceptual underpinnings for the analysis, and a longitudinal empirical analysis is conducted for an eight-year study period. The results indicate that not-for-profit hospitals with strong financial performance provide more unprofitable services for the insured and uninsured than do not-for-profit hospitals with weaker condition. For-profit hospital provision of these services is not influenced by their financial condition and instead may reflect actions to meet community expectations or to offer a sufficiently broad service array to maintain the business of insured patients. PMID- 21625343 TI - Investigations of the {ReO} core: A '2+2' complex from bidentate and potentially trident ligands: [ReO(eta-HOC(6)H(4)-2-CH(2)NC(6)H(4)S)(eta-SC(5)H(4)N)(PPh(3))]. AB - The reaction of [ReOCl(3)(PPh(3))(2)] with N-(2-hydroxybenzyl)-2-mercaptoaniline (H(3)hbma) (2) and 2-mercaptopyridine in hot CHCl yields [ReO(eta(2)-HOC(6)H(4)-2 CH(2)NC(6)H(4)S)(eta(2)-SC(5)H(4)N)(PPh(3))] (3). The structure of 3 consists of distorted octahedral Re(V) monomers. The coordination geometry at the rhenium is defined by a terminal oxo-group, the nitrogen and sulfur donors of the chelating mercaptopyridine, the nitrogen and sulfur donors of a bidentate (Hhbma)(2-) ligand, and the phosphorus of the PPh(3) group. The -C(6)H(4)OH arm of (Hhbma)(2 ) is pendant, and the coordinated nitrogen of this ligand is present as a deprotonated amido nitrogen. PMID- 21625344 TI - Bio-imaging, detection and analysis by using nanostructures as SERS substrates. AB - Surface-enhanced Raman scattering (SERS) is a phenomenon that occurs on nanoscale roughed metallic surface. The magnitude of the Raman scattering signal can be greatly enhanced when the scatterer is placed in the very close vicinity of the surface, which enables this phenomenon to be a highly sensitive analytical technique. SERS inherits the general strongpoint of conventional Raman spectroscopy and overcomes the inherently small cross section problem of a Raman scattering. It is a sensitive and nondestructive spectroscopic method for biological samples, and can be exploited either for the delivery of molecular structural information or for the detection of trace levels of analytes. Therefore, SERS has long been regarded as a powerful tool in biomedical research. Metallic nanostructure plays a key role in all the biomedical applications of SERS because the enhanced Raman signal can only be obtained on the surface of a finely divided substrate. This review focuses on progress made in the use of SERS as an analytical technique in bio-imaging, analysis and detection. Recent progress in the fabrication of SERS active nanostructures is also highlighted. PMID- 21625345 TI - Genetic Determinants of Virulence in the Marine Fish Pathogen Vibrio anguillarum. AB - One of the most studied fish pathogens is Vibrio anguillarum. Development of the genetics and biochemistry of the mechanisms of virulence in this fish pathogen together with clinical and ecologic studies has permitted the intensive development of microbiology in fish diseases. It is the intention of this review to compile the exhaustive knowledge accumulated on this bacterium and its interaction with the host fish by reporting a complete analysis of the V. anguillarum virulence factors and the genetics of their complexity. PMID- 21625346 TI - Viewing Low-Income Fathers' Ties to Families through a Cultural Lens: Insights for Research and Policy. AB - Policy makers have become increasingly interested in addressing the cultural dimensions of child support, "responsible fatherhood," and marriage in poor communities. However, policy studies have primarily focused on identifying economic determinants of these issues, with a substantial amount of variation in their statistical models left unexplained. This article draws on in-depth interviews the author conducted with disadvantaged mothers and fathers to illustrate how a systematic investigation into the meaning of low-income men's ties to families may fill in or provide alternative explanations for some important questions related to paternal involvement. In particular, it suggests that analyzing fathers' relationships through a cultural lens may not only reveal new information about the meaning of their emotional involvement, informal support, care of children, and conflicts with mothers which future policy studies should consider but may also inform policy initiatives by reducing the risk that they will be misdirected or have unintended consequences for poor families. PMID- 21625347 TI - A Constant-Volume Ventilator and Gas Recapture System for Hyperpolarized Gas MRI of Mouse and Rat Lungs. PMID- 21625348 TI - STATISTICAL INFERENCE OF TRANSMISSION FIDELITY OF DNA METHYLATION PATTERNS OVER SOMATIC CELL DIVISIONS IN MAMMALS. AB - We develop Bayesian inference methods for a recently-emerging type of epigenetic data to study the transmission fidelity of DNA methylation patterns over cell divisions. The data consist of parent-daughter double-stranded DNA methylation patterns with each pattern coming from a single cell and represented as an unordered pair of binary strings. The data are technically difficult and time consuming to collect, putting a premium on an efficient inference method. Our aim is to estimate rates for the maintenance and de novo methylation events that gave rise to the observed patterns, while accounting for measurement error. We model data at multiple sites jointly, thus using whole-strand information, and considerably reduce confounding between parameters. We also adopt a hierarchical structure that allows for variation in rates across sites without an explosion in the effective number of parameters. Our context-specific priors capture the expected stationarity, or near-stationarity, of the stochastic process that generated the data analyzed here. This expected stationarity is shown to greatly increase the precision of the estimation. Applying our model to a data set collected at the human FMR1 locus, we find that measurement errors, generally ignored in similar studies, occur at a non-trivial rate (inappropriate bisulfite conversion error: 1.6% with 80% CI: 0.9-2.3%). Accounting for these errors has a substantial impact on estimates of key biological parameters. The estimated average failure of maintenance rate and daughter de novo rate decline from 0.04 to 0.024 and from 0.14 to 0.07, respectively, when errors are accounted for. Our results also provide evidence that de novo events may occur on both parent and daughter strands: the median parent and daughter de novo rates are 0.08 (80% CI: 0.04-0.13) and 0.07 (80% CI: 0.04-0.11), respectively. PMID- 21625349 TI - Caveolae and lipid trafficking in adipocytes. AB - The abundance of caveolae in adipocytes suggests a possible cell-specific role for these structures, and because these cells take up and release fatty acids as their quantitatively most robust activity, modulation of fatty acid movement is one such role that is supported by substantial in vitro and in vivo data. In addition, caveolae are particularly rich in cholesterol and sphingolipids, and indeed, fat cells harbor more cholesterol than any other tissue. In this article, we review the role of adipocyte caveolae with regard to these important lipid classes. PMID- 21625350 TI - Assimilation and Individual Differences in Emotion: The Dynamics of Anger and Approach Motivation. AB - Individuals who cross cultural boundaries face many challenges when trying to adapt to a receiving culture. Adaptation challenges such as learning to maneuver across societal domains may become increasingly complex if structural level factors such as discrimination are present. Researchers have conceptualized acculturation as a relatively autonomous decision indicating that four acculturation strategies exist: assimilation, separation, integration, and marginalization. Moreover, researchers have also long debated the link between acculturation strategy, adaptation hassles and negative health outcomes. However, models seeking to explain how individual difference and structural level variables may influence each other and subsequently influence acculturation and adaptation are needed. The purpose of this study is to lay the foundation for the conceptualization of such a model. We propose that temperamental predispositions to negative emotionality, anger, and impulsivity may highlight discrimination which in turn may lead to increases in acculturative stress and negative markers of psychosocial well-being. We used SEM to test our hypothesized model. Results supported a modified model. Implications for the measurement of adaptation and interventions are discussed. PMID- 21625351 TI - Applicability of the Social Development Model to Urban Ethnic Minority Youth: Examining the Relationship between External Constraints, Family Socialization, and Problem Behaviors. AB - The development of preventive interventions targeting adolescent problem behaviors requires a thorough understanding of risk and protective factors for such behaviors. However, few studies examine whether different cultural and ethnic groups share these factors. This study is an attempt to fill a gap in research by examining similarities and differences in risk factors across racial and ethnic groups. The social development model has shown promise in organizing predictors of problem behaviors. This article investigates whether a version of that model can be generalized to youth in different racial and ethnic groups (N = 2,055, age range from 11 to 15), including African American (n = 478), Asian Pacific Islander (API) American (n = 491), multiracial (n = 442), and European American (n = 644) youth. The results demonstrate that common risk factors can be applied to adolescents, regardless of their race and ethnicity. The findings also demonstrate that there are racial and ethnic differences in the magnitudes of relationships among factors that affect problem behaviors. Further study is warranted to develop a better understanding of these differential magnitudes. PMID- 21625352 TI - To Model or Not to Model? A Dialogue on the Role of Computational Modeling in Developmental Science. AB - All sciences use models of some variety to understand complex phenomena. In developmental science, however, modeling is mostly limited to linear, algebraic descriptions of behavioral data. Some researchers have suggested that complex mathematical models of developmental phenomena are a viable (even necessary) tool that provide fertile ground for developing and testing theory as well as for generating new hypotheses and predictions. This paper explores the concerns, attitudes, and historical trends that underlie the tension between two cultures: one in which computational simulations of behavior are an important complement to observation and experimentation, and another which emphasizes evidence from behavioral experiments and linear models enhanced by verbal descriptions. This tension is explored as a dialogue between three characters: Ed (Experimental Developmentalist), Mira (Modeling Inclusive Research Advocate), and Phil (Philosopher of Science). PMID- 21625353 TI - Kidney/Bone Marrow Transplantation. PMID- 21625354 TI - THE USE OF HETEROLOGOUS ANTILYMPHOCYTE GLOBULIN (ALG) IN HUMAN RENAL AND LIVER TRANSPLANTATION. PMID- 21625355 TI - Hepatic Disorders in Renal Homograft Recipients. PMID- 21625356 TI - Modeling of Anatomical Information in Clustering of White Matter Fiber Trajectories Using Dirichlet Distribution. AB - In this work, we describe a white matter trajectory clustering algorithm that allows for incorporating and appropriately weighting anatomical information. The influence of the anatomical prior reflects confidence in its accuracy and relevance. It can either be defined by the user or it can be inferred automatically. After a detailed description of our novel clustering framework, we demonstrate its properties through a set of preliminary experiments. PMID- 21625357 TI - Outcome after Liver Transplantation for Cystic Fibrosis. PMID- 21625358 TI - Bias in Hotelling Observer Performance Computed from Finite Data. AB - An observer performing a detection task analyzes an image and produces a single number, a test statistic, for that image. This test statistic represents the observers "confidence" that a signal (e.g., a tumor) is present. The linear observer that maximizes the test-statistic SNR is known as the Hotelling observer. Generally, computation of the Hotelling SNR, or Hotelling trace, requires the inverse of a large covariance matrix. Recent developments have resulted in methods for the estimation and inversion of these large covariance matrices with relatively small numbers of images. The estimation and inversion of these matrices is made possible by a covariance-matrix decomposition that splits the full covariance matrix into an average detector-noise component and a background-variability component. Because the average detector-noise component is often diagonal and/or easily estimated, a full-rank, invertible covariance matrix can be produced with few images. We have studied the bias of estimates of the Hotelling trace using this decomposition for high-detector-noise and low-detector noise situations. In extremely low-noise situations, this covariance decomposition may result in a significant bias. We will present a theoretical evaluation of the Hotelling-trace bias, as well as extensive simulation studies. PMID- 21625359 TI - Evaluation of hardware in a small-animal SPECT system using reconstructed images. AB - Evaluation of imaging hardware represents a vital component of system design. In small-animal SPECT imaging, this evaluation has become increasingly difficult with the emergence of multi-pinhole apertures and adaptive, or patient-specific, imaging. This paper will describe two methods for hardware evaluation using reconstructed images. The first method is a rapid technique incorporating a system-specific non-linear, three-dimensional point response. This point response is easily computed and offers qualitative insight into an aperture's resolution and artifact characteristics. The second method is an objective assessment of signal detection in lumpy backgrounds using the channelized Hotelling observer (CHO) with 3D Laguerre-Gauss and difference-of-Gaussian channels to calculate area under the receiver-operating characteristic curve (AUC). Previous work presented at this meeting described a unique, small-animal SPECT system (M(3)R) capable of operating under a myriad of hardware configurations and ideally suited for image quality studies. Measured system matrices were collected for several hardware configurations of M(3)R. The data used to implement these two methods was then generated by taking simulated objects through the measured system matrices. The results of these two methods comprise a combination of qualitative and quantitative analysis that is well-suited for hardware assessment. PMID- 21625360 TI - "Yes, I Do But Not With You"-Qualitative Analyses of Sexual/Romantic Overture related Aggression in Bars and Clubs. AB - Assaultive and aggressive behaviors related to sexual overtures are common in commercial drinking establishments (bars, pubs and clubs). In this paper, we examined the thematic content of 251 incidents of verbal and physical aggression related to sexual/romantic overtures documented by researcher-observers in a study of Toronto bar and clubs. Aggression was examined as it emerged in the following stages of the social interaction process: (a) sexual/romantic overtures that began aggressively; (b) initiators of sexual/romantic overtures who became aggressive later in the social interaction process; (c) aggression by targets of overtures; and (d) aggression by third parties. From these thematic analyses, we identify the distinctions between predatory and genuine overtures and explore the potential role of the effects of alcohol.In these social overtures, aggression occurred as part of the initial overture, during the interaction following the overture (i.e., aggression by the person who made the initial overture, by the target or third parties) and in response to rejection by the target. Targets of overtures responded aggressively to perceived inappropriate overtures; third parties played important aggressive and nonaggressive roles; and alcohol intoxication was identified as contributing to aggression in a number of ways. The theoretical significance and practical implications for prevention of the findings are discussed. PMID- 21625361 TI - On the Binding Strength Sequence for Nucleic Acid Bases and C(60) with Density Functional and Dispersion-corrected Density Functional Theories: Whether C(60) could protect nucleic acid bases from radiation-induced damage? AB - The major objective of this paper is to address a controversial binding sequence between nucleic acid bases (NABs) and C(60) by investigating adsorptions of NABs and their cations on C(60) fullerene with a variety of density functional theories including two novel hybrid meta-GGA functionals, M05-2x and M06-2x, as well as a dispersion-corrected density functional, PBE-D. The M05-2x/6-311++G** provides the same binding sequence as previously reported, guanine(G) > cytosine(C) > adenine (A) > thymine (T); however, M06-2x switches the binding strengths of A and C, and PBE-D eventually results in the following sequence, G>A>T>C, which is the same as the widely accepted hierarchy for the stacking of NABs on other carbon nanomaterials such as single-walled carbon nanotube and graphite. The results indicate that the questionable relative binding strength is due to insufficient electron correlation treatment with the M05-2x or even the M06-2x method. The binding energy of G@C(60) obtained with the M06-2x/6 311++G(d,p) and the PBE-D/cc-pVDZ is -7.10 and -8.07 kcal/mol, respectively, and the latter is only slightly weaker than that predicted by the MP2/6-31G(d,p) ( 8.10kca/mol). Thus, the PDE-D performs better than the M06-2x for the observed NAB@C(60) pi-stacked complexes. To discuss whether C(60) could prevent NABs from radiation-induced damage, ionization potentials of NABs and C(60), and frontier molecular orbitals of the complexes NABs@C(60) and (NABs@C(60))(+) are also extensively investigated. These results revealed that when an electron escapes from the complexes, a hole was preferentially created in C(60) for T and C complexes, while for G and A the hole delocalizes over the entire complex, rather than a localization on the C(60) moiety. The interesting finding might open a new strategy for protecting DNA from radiation-induced damage and offer a new idea for designing C(60)-based antiradiation drugs. PMID- 21625362 TI - Specificity and signaling in the Drosophila immune response. AB - The Drosophila immune response is characterized by the rapid and robust production of a battery of antimicrobial peptides immediately following infection. The genes encoding these antimicrobial peptides are controlled by two NF-kappaB signaling pathways that respond to microbial infection. The IMD pathway is triggered by DAP-type peptidoglycan, from the cell wall of most Gram-negative and certain Gram-positive bacteria, and activates the NF-kappaB precursor protein Relish. The Toll pathway, on the other hand, is stimulated by lysine-type peptidoglycan from many Gram-positive bacteria, beta 1,3 glucans from many fungi, as well as by microbial proteases. Toll signaling leads to the activation and nuclear translocation of DIF or Dorsal, two other NF-kappaB homologs. This review presents our current understanding of the molecular mechanisms involved in microbial recognition and signal transduction in these two innate immune pathways. PMID- 21625363 TI - On the origin of a domesticated species: Identifying the parent population of Russian silver foxes (Vulpes vulpes). AB - The foxes at Novosibirsk, Russia, are the only population of domesticated foxes in the world. These domesticated foxes originated from farm-bred silver foxes (Vulpes vulpes), whose genetic source is unknown. In this study we examined the origin of the domesticated strain of foxes and two other farm-bred fox populations (aggressive and unselected) maintained in Novosibirsk. To identify the phylogenetic origin of these populations we sequenced two regions of mtDNA, cytochrome b and D-loop, from 24 Novosibirsk foxes (8 foxes from each population) and compared them with corresponding sequences of native red foxes from Europe, Asia, Alaska and Western Canada, Eastern Canada, and the Western Mountains of the USA. We identified seven cytochrome b - D-loop haplotypes in Novosibirsk populations, four of which were previously observed in Eastern North America. The three remaining haplotypes differed by one or two base change from the most common haplotype in Eastern Canada. Phi(ST) analysis showed significant differentiation between Novosibirsk populations and red fox populations from all geographic regions except Eastern Canada. No haplotypes of Eurasian origin were identified in the Novosibirsk populations. These results are consistent with historical records indicating that the original breeding stock of farm-bred foxes originated from Prince Edward Island, Canada. Mitochondrial DNA data together with historical records indicate two stages in the selection of domesticated foxes: the first includes captive breeding for ~50 years with unconscious selection for behaviour; the second corresponds to over 50 further years of intensive selection for tame behaviour. PMID- 21625364 TI - A Scanning Transmission Electron Microscopy Method for Determining Manganese Composition in Welding Fume as a Function of Primary Particle Size. AB - Increasing evidence suggests that the physicochemical properties of inhaled nanoparticles influence the resulting toxicokinetics and toxicodynamics. This report presents a method using scanning transmission electron microscopy (STEM) to measure the Mn content throughout the primary particle size distribution of welding fume particle samples collected on filters for application in exposure and health research. Dark field images were collected to assess the primary particle size distribution and energy-dispersive X-ray and electron energy loss spectroscopy were performed for measurement of Mn composition as a function of primary particle size. A manual method incorporating imaging software was used to measure the primary particle diameter and to select an integration region for compositional analysis within primary particles throughout the size range. To explore the variation in the developed metric, the method was applied to 10 gas metal arc welding (GMAW) fume particle samples of mild steel that were collected under a variety of conditions. The range of Mn composition by particle size was 0.10 to 0.19 %/nm, where a positive estimate indicates greater relative abundance of Mn increasing with primary particle size and a negative estimate conversely indicates decreasing Mn content with size. However, the estimate was only statistically significant (p<0.05) in half of the samples (n=5), which all had a positive estimate. In the remaining samples, no significant trend was measured. Our findings indicate that the method is reproducible and that differences in the abundance of Mn by primary particle size among welding fume samples can be detected. PMID- 21625365 TI - Kaempferol induces apoptosis in ovarian cancer cells through activating p53 in the intrinsic pathway. AB - Ovarian cancer is a significant malignancy for women in the western world, and its death rate has remained unchanged over the past 50 years, leaving room for proper chemoprevention. Kaempferol is a natural flavonoid widely distributed in fruits and vegetables, and epidemiological studies have found a negative correlation between kaempferol consumption and ovarian cancer risk. To understand the mechanism behind this negative correlation, we investigated kaempferol's ability to induce apoptosis in A2780/CP70, A2780/wt, and OVCAR-3 ovarian cancer cell lines. Kaempferol inhibited cell proliferation but did not cause necrosis in all 3 cell lines. For the apoptosis, caspase 3/7 levels were induced in a concentration-dependent manner by kaempferol treatment, with A2780/wt cells being the most responsive. This induction can be diminished by pre-treatment with a caspase-9 inhibitor, indicating an intrinsic apoptosis pathway. Western blot analysis revealed that protein levels of Bcl-x(L) were decreased in ovarian cancer cells, while p53, Bad, and Bax proteins were up-regulated by kaempferol treatment. Our data indicate that kaempferol induces apoptosis in ovarian cancer cells through regulating pro-apoptotic and anti-apoptotic protein expressions in the intrinsic apoptosis pathways, and is a good candidate for the chemoprevention of ovarian cancers in humans. Further studies in animal models and clinical trials are therefore warranted. PMID- 21625366 TI - Sparse Regulatory Networks. AB - In many organisms the expression levels of each gene are controlled by the activation levels of known "Transcription Factors" (TF). A problem of considerable interest is that of estimating the "Transcription Regulation Networks" (TRN) relating the TFs and genes. While the expression levels of genes can be observed, the activation levels of the corresponding TFs are usually unknown, greatly increasing the difficulty of the problem. Based on previous experimental work, it is often the case that partial information about the TRN is available. For example, certain TFs may be known to regulate a given gene or in other cases a connection may be predicted with a certain probability. In general, the biology of the problem indicates there will be very few connections between TFs and genes. Several methods have been proposed for estimating TRNs. However, they all suffer from problems such as unrealistic assumptions about prior knowledge of the network structure or computational limitations. We propose a new approach that can directly utilize prior information about the network structure in conjunction with observed gene expression data to estimate the TRN. Our approach uses L(1) penalties on the network to ensure a sparse structure. This has the advantage of being computationally efficient as well as making many fewer assumptions about the network structure. We use our methodology to construct the TRN for E. coli and show that the estimate is biologically sensible and compares favorably with previous estimates. PMID- 21625367 TI - Heterogeneity in returns to college education: selection bias in contemporary taiwan. AB - The causal impact of higher education on earnings may be heterogeneous across different members of a population. Using a newly developed instrumental-variable method in economics, we illustrate heterogeneous treatment effects of higher education on earnings resulting from sorting mechanisms that select individuals with certain unobserved attributes into college education. The setting of our empirical work is contemporary Taiwan -- a transitional economy that has recently experienced a rapid expansion in higher education. We find distinct patterns by gender, with selection bias most clearly shown among women but not among men: the college return to earnings is on average greater for women who actually attended college than women who did not attend college. PMID- 21625368 TI - Metropolitan influences on migration into poor and nonpoor neighborhoods. AB - Data from the Panel Study of Income Dynamics and three decennial U.S. censuses are used to examine the influence of metropolitan-area characteristics on black and white households' propensity to move into poor versus nonpoor neighborhoods. We find that a nontrivial portion of the variance in the odds of moving to a poor rather to a nonpoor neighborhood exists between metropolitan areas. Net of established individual-level predictors of inter-neighborhood migration, black and white households are more likely to move to a poor or extremely poor tract rather than to a nonpoor tract in metropolitan areas containing many poor neighborhoods and a paucity of recently-built housing in nonpoor areas. Blacks are especially likely to move to a poor tract in metropolitan areas characterized by high levels of racial residential segregation and in which poor tracts have a sizeable concentration of blacks. White households are more likely to move to a poor than to a nonpoor tract in metropolitan areas that have comparatively few African Americans. PMID- 21625369 TI - New method for separation of electrode polarization impedance from measured tissue impedance. AB - In this paper we have shown that electrode polarization impedance (EPI) can be separated from measured tissue impedance as long as the characteristic frequencies of EPI and tissue are not too close, so that the EPI is largely displayed as a separate dispersion. In 2-electrode measurements the EPI and sample are physically connected in series, and commonly modelled by equivalent components in series. We have calculated the parallel equivalent elements and converted the series connected EPI and sample to a parallel admittance model. By curve fitting on the converted model we have shown that this provides a new method for estimating the EPI with enhanced accuracy compared to similar techniques used on the impedance model. PMID- 21625371 TI - Conventions for sign and speech transcription of child bimodal bilingual corpora in ELAN. AB - This article extends current methodologies for the linguistic analysis of sign language acquisition to cases of bimodal bilingual acquisition. Using ELAN, we are transcribing longitudinal spontaneous production data from hearing children of Deaf parents who are learning either American Sign Language (ASL) and American English (AE), or Brazilian Sign Language (Libras, also referred to as Lingua de Sinais Brasileira/LSB in some texts) and Brazilian Portuguese (BP). Our goal is to construct corpora that can be mined for a wide range of investigations on various topics in acquisition. Thus, it is important that we maintain consistency in transcription for both signed and spoken languages. This article documents our transcription conventions, including the principles behind our approach. Using this document, other researchers can chose to follow similar conventions or develop new ones using our suggestions as a starting point. PMID- 21625370 TI - Barriers to predicting the mechanisms and risk factors of non-contact anterior cruciate ligament injury. AB - High incidences of non-contact anterior cruciate ligament (ACL) injury, frequent requirements for ACL reconstruction, and limited understanding of ACL mechanics have engendered considerable interest in quantifying the ACL loading mechanisms. Although some progress has been made to better understand non-contact ACL injuries, information on how and why non-contact ACL injuries occur is still largely unavailable. In other words, research is yet to yield consensus on injury mechanisms and risk factors. Biomechanics, video analysis, and related study approaches have elucidated to some extent how ACL injuries occur. However, these approaches are limited because they provide estimates, rather than precise measurements of knee - and more specifically ACL - kinematics at the time of injury. These study approaches are also limited in their inability to simultaneously capture many of the contributing factors to injury.This paper aims at elucidating and summarizing the key challenges that confound our understanding in predicting the mechanisms and subsequently identifying risk factors of non contact ACL injury. This work also appraise the methodological rigor of existing study approaches, review testing protocols employed in published studies, as well as presents a possible coupled approach to better understand injury mechanisms and risk factors of non-contact ACL injury. Three comprehensive electronic databases and hand search of journal papers, covering numerous full text published English articles were utilized to find studies on the association between ACL and injury mechanisms, ACL and risk factors, as well as, ACL and investigative approaches. This review unveils that new research modalities and/or coupled research methods are required to better understand how and why the ACL gets injured. Only by achieving a better understanding of ACL loading mechanisms and the associated contributing factors, one will be able to develop robust prevention strategies and exercise regimens to mitigate non-contact ACL injuries. PMID- 21625372 TI - A Rank-Based Test for Comparison of Multidimensional Outcomes. AB - For comparison of multiple outcomes commonly encountered in biomedical research, Huang et al. (2005) improved O'Brien's (1984) rank-sum tests through the replacement of the ad hoc variance by the asymptotic variance of the test statistics. The improved tests control the Type I error rate at the desired level and gain power when the differences between the two comparison groups in each outcome variable fall into the same direction. However, they may lose power when the differences are in different directions (e.g., some are positive and some are negative). These tests and the popular Bonferroni correction failed to show important significant difference when applied to compare heart rates from a clinical trial to evaluate the effect of a procedure to remove the cardioprotective solution HTK. We propose an alternative test statistic, taking the maximum of the individual rank-sum statistics, which controls the type I error and maintains satisfactory power regardless of the directions of the differences. Simulation studies show the proposed test to be of higher power than other tests in certain alternative parameter space of interest. Furthermore, when used to analyze the heart rates data the proposed test yields more satisfactory results. PMID- 21625373 TI - Interactions of human endothelial and multipotent mesenchymal stem cells in cocultures. AB - Current strategies for tissue engineering of bone rely on the implantation of scaffolds, colonized with human mesenchymal stem cells (hMSC), into a recipient. A major limitation is the lack of blood vessels. One approach to enhance the scaffold vascularisation is to supply the scaffolds with endothelial cells (EC).The main goal of this study was to establish a coculture system of hMSC and EC for the purposes of bone tissue engineering. Therefore, the cell behaviour, proliferation and differentiation capacity in various cell culture media as well as cell interactions in the cocultures were evaluated.The differentiation capacity of hMSC along osteogenic, chondrogenic, and adipogenic lineage was impaired in EC medium while in a mixed EC and hMSC media, hMSC maintained osteogenic differentiation. In order to identify and trace EC in the cocultures, EC were transduced with eGFP. Using time-lapse imaging, we observed that hMSC and EC actively migrated towards cells of their own type and formed separate clusters in long term cocultures. The scarcity of hMSC and EC contacts in the cocultures suggest the influence of growth factor-mediated cell interactions and points to the necessity of further optimization of the coculture conditions. PMID- 21625374 TI - Pharmacokinetic-Pharmacodynamic Modeling of Metformin for the Treatment of Type II Diabetes Mellitus. AB - Metformin is an antihyperglycemic agent commonly used for the treatment of Type II diabetes mellitus. However, its effects on patients are derived usually from clinical experiments. In this study, a dynamic model of Type II diabetes mellitus with the treatment of metformin is proposed. The Type II diabetic model is a modification of an existing compartmental diabetic model. The dynamic simulation of the metformin effect for a Type II diabetic patient is based on the pharmacokinetic and pharmacodynamic relationship with a human body. The corresponding model parameters are estimated by optimization using clinical data from published reports. Then, the effect of metformin in both intravenous and oral administration on a Type II diabetes mellitus model are compared. The combination treatment of insulin infusion plus oral metformin is shown to be superior than the monotherapy with oral metformin only. These results are consistent with the clinical understanding of the use of metformin. For further work, the model can be analyzed for evaluating the treatment of diabetes mellitus with different pharmacological agents. PMID- 21625375 TI - Nano-material aspects of shock absorption in bone joints. AB - This theoretical study is based on a nano-technological evaluation of the effect of pressure on the composite bone fine structure. It turned out, that the well known macroscopic mechano-elastic performance of bones in combination with muscles and tendons is just one functional aspect which is critically supported by additional micro- and nano- shock damping technology aimed at minimising local bone material damage within the joints and supporting spongy bone material. The identified mechanisms comprise essentially three phenomena localised within the three-dimensional spongy structure with channels and so called perforated flexible tensulae membranes of different dimensions intersecting and linking them. Kinetic energy of a mechanical shock may be dissipated within the solid liquid composite bone structure into heat via the generation of quasi-chaotic hydromechanic micro-turbulence. It may generate electro-kinetic energy in terms of electric currents and potentials. And the resulting specific structural and surface electrochemical changes may induce the compressible intra-osseal liquid to build up pressure dependent free chemical energy. Innovative bone joint prostheses will have to consider and to be adapted to the nano-material aspects of shock absorption in the operated bones. PMID- 21625376 TI - Comparison of TCP and TCP/HA Hybrid Scaffolds for Osteoconductive Activity. AB - Two types of porous ceramic scaffolds were prepared, consisting of beta tricalcium phosphate (TCP) or the mixed powder of TCP and hydroxyapatite (HA) at a 2:1 mass ratio. A variety of methods have been used to fabricate bone scaffolds, while the sintering approach was adopted in this work. An extremely high temperature was used on sintering that proposed to consolidate the ceramic particles. As revealed by SEM, a well opened pore structure was developed within the scaffolds. The theta-values were measured to be of 73.3 degrees and 6.5 degrees for the composite scaffold and TCP sample, respectively. According to XRD patterns, the existence of grains coalescence and partial bonding between HA and TCP powders was demonstrated. Scaffold mechanical property in the term of flexural strength was also determined. The result showed decreasing of the strength by HA supplement, suggesting the more brittle characteristic of HA in comparison with TCP. By soaking the composite scaffold in PBS for a period of 2 weeks, transformation from particles to flank-like crystalline was clearly observed. Such change was found to be favorable for cell attachment, migration, and growth. By implanting cell-seeded scaffolds into nude mice, an abundant osseous extracellular matrix was identified for the composite implants. In contrast, the matrix was minimally detected in TCP implanted samples. Thus, the composite scaffold was found superior for hard tissue regeneration. PMID- 21625377 TI - Evaluation of madurahydroxylactone as a slow release antibacterial implant coating. AB - Madurahydroxylactone (MHL), a secondary metabolite with antibacterial activity was evaluated for its suitability to generate controlled drug release coatings on medical implant materials. A smooth and firmly attached layer could be produced from a precursor solution on various metallic implant materials. In physiological salt solutions these coatings dissolved within a time period up to one week. A combination of MHL with a broad spectrum fluoroquinolone antibiotic was used to create a coating that was active against all bacterial strains tested. The time period during which the coating remained active against Pseudomonas aeruginosa was investigated. The results indicated a delayed drug release from single layer coatings in the course of seven days. MHL was biocompatible in cell culture assays and could after a delay even serve as a cell adhesion substrate for human or murine cells. The findings indicate a potential for MHL for the generation of delayed release antimicrobial implant coatings. PMID- 21625378 TI - Identifying risk factors for marijuana use among veterans affairs patients. AB - OBJECTIVES: Cannabis is the most widely used drug in the United States, and its use carries negative health consequences; however, universal screening for cannabis use is cumbersome. If data commonly collected in the primary care setting (eg, use of alcohol, smoking status, and depression symptoms) could predict cannabis use, then providers can implement targeted marijuana screening in high-risk groups. METHODS: We reviewed Behavioral Health Laboratory data collected between 2003 and 2006 from 5512 patients referred by Veterans Affairs primary care clinics for potential mental health needs. Logistic regression was used to determine the predictors of past year marijuana use. RESULTS: A total of 11.5% of the sample reported using marijuana in the past year. Age, gender, other drug use, presence of alcohol use disorders, smoking status, depressive disorders, posttraumatic stress disorder, anxiety disorders, and psychotic symptoms, individually, were associated with the patients' use of marijuana during the past year. When controlling for age, race, and gender in a logistic regression analyses, only other drug use, alcohol use disorder, and smoking status were linked to past year marijuana use. Patients were 5.4 (95% confidence interval [CI] 4.3-6.7) times more likely to have used marijuana during the past year if they used another illicit drug during the past year. Those with alcohol use disorder diagnosis or current smokers were 2.3 (95% CI 1.9-2.8) and 1.5 times (95% CI 1.3-1.7), respectively, more likely to have used marijuana during the past year. Receiver operating characteristic curve (area under curve = 0.79) represents good sensitivity and specificity of the model, correctly classifying 88.4% of the past year marijuana users. CONCLUSION: Identifying patients at high risk for cannabis use may facilitate targeted screening and provision of interventions in primary care. Patients who screen positive for cigarette use, alcohol abuse or dependence, or have evidence of other illicit drug use could be considered for cannabis screening. PMID- 21625379 TI - Competitive Binding to Cuprous Ions of Protein and BCA in the Bicinchoninic Acid Protein Assay. AB - Although Bicinchoninic acid (BCA) has been widely used to determine protein concentration, the mechanism of interaction between protein, copper ion and BCA in this assay is still not well known. Using the Micro BCA protein assay kit (Pierce Company), we measured the absorbance at 562 nm of BSA solutions with different concentrations of protein, and also varied the BCA concentration. When the concentration of protein was increased, the absorbance exhibited the known linear and nonlinear increase, and then reached an unexpected plateau followed by a gradual decrease. We introduced a model in which peptide chains competed with BCA for binding to cuprous ions. Formation of the well-known chromogenic complex of BCA-Cu(1+)-BCA was competed with the binding of two peptide bonds (NTPB) to cuprous ion, and there is the possibility of the existence of two new complexes. A simple equilibrium equation was established to describe the correlations between the substances in solution at equilibrium, and an empirical exponential function was introduced to describe the reduction reaction. Theoretical predictions of absorbance from the model were in good agreement with the measurements, which not only validated the competitive binding model, but also predicted a new complex of BCA-Cu(1+)-NTPB that might exist in the final solution. This work provides a new insight into understanding the chemical bases of the BCA protein assay and might extend the assay to higher protein concentration. PMID- 21625380 TI - Safer Sexual Practices and HIV Screening Behavior among Rural California American Indians. AB - This paper reports on safer sexual practices and HIV screening behavior among rural California American Indians. Thirteen Indian health clinic registries formed the random household survey sampling frame (N=457). Measures included socio-demographics, safer sexual practices, HIV testing, high-risk behaviors, perception of wellness, general health status, neglect, physical and sexual abuse history. Statistical tests included chi-square and Fisher's exact tests, as well as multiple logistic regression analysis. Respondents who practiced safer sex were younger, not married, reported lower annual income, changed sexual behavior in the last 12 months, were not in a monogamous relationship, and had a higher perception of wellness. Those who were tested for HIV were younger, had at least a high school education, consumed alcohol, had significantly higher suicide ideation and attempts, changed their sexual behavior in the last 12 months, and reported a history of sexual abuse as a child and adult. Very little is known about HIV prevention and screening behaviors in rural American Indian populations. Identifying predictors of unsafe sexual practices and non-HIV testing tendencies help to plan for measures to protect the population from this devastating disease. We make recommendations for implementing HIV prevention education and screenings at American Indian clinics. PMID- 21625381 TI - California Wellness Study: American Indians and Obesity. AB - This paper identifies the prevalence and predictors of obesity among California's American Indian adults. A cross-sectional study was conducted at 13 rural sites. Indian healthcare clinics served as the sampling frame and were selected because of their proximity and access to the target population. Four-hundred and fifty adult American Indians participated; 74 percent were female and 26 percent were male. The average age was 40, ranging from 18-74. Measures included socio demographics, general health, BMI, type 2 diabetes, exercise and dietary habits. Eighty-two percent were overweight, obese or morbidly obese. Chi-square tests revealed three variables significantly associated with BMI categories: having type 2 diabetes, female gender and poor general health status. A logistic regression model for obese/morbidly obese (BMI > 30) versus overweight/normal (BMI < 30) persons found gender and diabetes status as significant predictors, while general health status showed trend. Females had 1.59 greater odds of being obese than males (p=0.04). Those that do not have diabetes are less likely to be obese (p=0.02). Those that do not have good general health were 2.5 times more likely to be obese than those that have good general health (p=0.06). Overall goodness of fit was significant (p=0.0009). It is important to identify individuals and population who are normal/overweight, obese/morbidly obese so support and interventions can be planned and implemented. PMID- 21625382 TI - Use of Videophone Technology to Address Medication Adherence Issues in Persons with HIV. AB - Adherence to HAART medication regimens is vital to maintaining suppression of HIV, but persons with HIV face many challenges to adhering consistently to HIV medication regimens. This is particularly true for persons who live in geographically-isolated areas or who have significant levels of cognitive compromise. A videophone-based version of Reynolds' HAART CARE (HC) telephone intervention for medication adherence was pilot-tested with 23 persons living with HIV residing in both urban and non-urban communities. The purpose of the pilot study was to evaluate the feasibility and acceptance of an adherence improvement intervention administered via videophones. Furthermore, the feasibility and acceptability of conducting HIV pill counts through videophones were assessed. The videophone version of HC produced significant increases in self-reported rates of medication adherence and was generally well-received by interventionists and participants. Pill counts conducted via videophone were also well-accepted by participants. Self-reported adherence levels were higher than videophone-based pill count adherence levels. Challenges to the use of videophones included the requirement that only analog landlines be used, poor quality of video and audio transmissions, and high cost for equipment. Methods to overcome these challenges are discussed. PMID- 21625384 TI - Evaluating the fidelity of de novo short read metagenomic assembly using simulated data. AB - A frequent step in metagenomic data analysis comprises the assembly of the sequenced reads. Many assembly tools have been published in the last years targeting data coming from next-generation sequencing (NGS) technologies but these assemblers have not been designed for or tested in multi-genome scenarios that characterize metagenomic studies. Here we provide a critical assessment of current de novo short reads assembly tools in multi-genome scenarios using complex simulated metagenomic data. With this approach we tested the fidelity of different assemblers in metagenomic studies demonstrating that even under the simplest compositions the number of chimeric contigs involving different species is noticeable. We further showed that the assembly process reduces the accuracy of the functional classification of the metagenomic data and that these errors can be overcome raising the coverage of the studied metagenome. The results presented here highlight the particular difficulties that de novo genome assemblers face in multi-genome scenarios demonstrating that these difficulties, that often compromise the functional classification of the analyzed data, can be overcome with a high sequencing effort. PMID- 21625383 TI - Glioblastoma models reveal the connection between adult glial progenitors and the proneural phenotype. AB - BACKGROUND: Tumor heterogeneity is a major obstacle for finding effective treatment of Glioblastoma (GBM). Based on global expression analysis, GBM can be classified into distinct subtypes: Proneural, Neural, Classical and Mesenchymal. The signatures of these different tumor subtypes may reflect the phenotypes of cells giving rise to them. However, the experimental evidence connecting any specific subtype of GBM to particular cells of origin is lacking. In addition, it is unclear how different genetic alterations interact with cells of origin in determining tumor heterogeneity. This issue cannot be addressed by studying end stage human tumors. METHODOLOGY/PRINCIPAL FINDINGS: To address this issue, we used retroviruses to deliver transforming genetic lesions to glial progenitors in adult mouse brain. We compared the resulting tumors to human GBM. We found that different initiating genetic lesions gave rise to tumors with different growth rates. However all mouse tumors closely resembled the human Proneural GBM. Comparative analysis of these mouse tumors allowed us to identify a set of genes whose expression in humans with Proneural GBM correlates with survival. CONCLUSIONS/SIGNIFICANCE: This study offers insights into the relationship between adult glial progenitors and Proneural GBM, and allows us to identify molecular alterations that lead to more aggressive tumor growth. In addition, we present a new preclinical model that can be used to test treatments directed at a specific type of GBM in future studies. PMID- 21625385 TI - Comparison of LFP-based and spike-based spectro-temporal receptive fields and cross-correlation in cat primary auditory cortex. AB - Multi-electrode array recordings of spike and local field potential (LFP) activity were made from primary auditory cortex of 12 normal hearing, ketamine anesthetized cats. We evaluated 259 spectro-temporal receptive fields (STRFs) and 492 frequency-tuning curves (FTCs) based on LFPs and spikes simultaneously recorded on the same electrode. We compared their characteristic frequency (CF) gradients and their cross-correlation distances. The CF gradient for spike-based FTCs was about twice that for 2-40 Hz-filtered LFP-based FTCs, indicating greatly reduced frequency selectivity for LFPs. We also present comparisons for LFPs band pass filtered between 4-8 Hz, 8-16 Hz and 16-40 Hz, with spike-based STRFs, on the basis of their marginal frequency distributions. We find on average a significantly larger correlation between the spike based marginal frequency distributions and those based on the 16-40 Hz filtered LFP, compared to those based on the 4-8 Hz, 8-16 Hz and 2-40 Hz filtered LFP. This suggests greater frequency specificity for the 16-40 Hz LFPs compared to those of lower frequency content. For spontaneous LFP and spike activity we evaluated 1373 pair correlations for pairs with >200 spikes in 900 s per electrode. Peak correlation coefficient space constants were similar for the 2-40 Hz filtered LFP (5.5 mm) and the 16-40 Hz LFP (7.4 mm), whereas for spike-pair correlations it was about half that, at 3.2 mm. Comparing spike-pairs with 2-40 Hz (and 16-40 Hz) LFP-pair correlations showed that about 16% (9%) of the variance in the spike-pair correlations could be explained from LFP-pair correlations recorded on the same electrodes within the same electrode array. This larger correlation distance combined with the reduced CF gradient and much broader frequency selectivity suggests that LFPs are not a substitute for spike activity in primary auditory cortex. PMID- 21625386 TI - PTHrP induces autocrine/paracrine proliferation of bone tumor cells through inhibition of apoptosis. AB - Giant Cell Tumor of Bone (GCT) is an aggressive skeletal tumor characterized by local bone destruction, high recurrence rates and metastatic potential. Previous work in our lab has shown that the neoplastic cell of GCT is a proliferating pre osteoblastic stromal cell in which the transcription factor Runx2 plays a role in regulating protein expression. One of the proteins expressed by these cells is parathyroid hormone-related protein (PTHrP). The objectives of this study were to determine the role played by PTHrP in GCT of bone with a focus on cell proliferation and apoptosis. Primary stromal cell cultures from 5 patients with GCT of bone and one lung metastasis were used for cell-based experiments. Control cell lines included a renal cell carcinoma (RCC) cell line and a human fetal osteoblast cell line. Cells were exposed to optimized concentrations of a PTHrP neutralizing antibody and were analyzed with the use of cell proliferation and apoptosis assays including mitochondrial dehydrogenase assays, crystal violet assays, APO-1 ELISAs, caspase activity assays, flow cytometry and immunofluorescent immunohistochemistry. Neutralization of PTHrP in the cell environment inhibited cell proliferation in a consistent manner and induced apoptosis in the GCT stromal cells, with the exception of those obtained from a lung metastasis. Cell cycle progression was not significantly affected by PTHrP neutralization. These findings indicate that PTHrP plays an autocrine/paracrine neoplastic role in GCT by allowing the proliferating stromal cells to evade apoptosis, possibly through non-traditional caspase-independent pathways. Thus PTHrP neutralizing immunotherapy is an intriguing potential therapeutic strategy for this tumor. PMID- 21625387 TI - The role of miR-103 and miR-107 in regulation of CDK5R1 expression and in cellular migration. AB - CDK5R1 encodes p35, a specific activator of the serine/threonine kinase CDK5, which plays crucial roles in CNS development and maintenance. CDK5 activity strongly depends on p35 levels and p35/CDK5 misregulation is deleterious for correct CNS function, suggesting that a tightly controlled regulation of CDK5R1 expression is needed for proper CDK5 activity. Accordingly, CDK5R1 expression was demonstrated to be controlled at both transcriptional and post-transcriptional levels, but a possible regulation through microRNAs (miRNAs) has never been investigated. We predicted, within the large CDK5R1 3'UTR several miRNA target sites. Among them, we selected for functional studies miR-103 and miR-107, whose expression has shown a strong inverse correlation with p35 levels in different cell lines. A significant reduction of CDK5R1 mRNA and p35 levels was observed after transfection of SK-N-BE neuroblastoma cells with the miR-103 or miR-107 precursor (pre-miR-103 or pre-miR-107). Conversely, p35 levels significantly increased following transfection of the corresponding antagonists (anti-miR-103 or anti-miR-107). Moreover, the level of CDK5R1 transcript shifts from the polysomal to the subpolysomal mRNA fraction after transfection with pre-miR-107 and, conversely, from the subpolysomal to the polysolmal mRNA fraction after transfection with anti-miR-107, suggesting a direct action on translation efficiency. We demonstrate, by means of luciferase assays, that miR-103 and miR 107 are able to directly interact with the CDK5R1 3'-UTR, in correspondence of a specific target site. Finally, miR-103 and miR-107 overexpression, as well as CDK5R1 silencing, caused a reduction in SK-N-BE migration ability, indicating that these miRNAs affect neuronal migration by modulating CDK5R1 expression. These findings indicate that miR-103 and miR-107 regulate CDK5R1 expression, allowing us to hypothesize that a miRNA-mediated mechanism may influence CDK5 activity and the associated molecular pathways. PMID- 21625388 TI - Connectivity within and among a network of temperate marine reserves. AB - Networks of marine reserves are increasingly being promoted as a means of conserving marine biodiversity. One consideration in designing systems of marine reserves is the maintenance of connectivity to ensure the long-term persistence and resilience of populations. Knowledge of connectivity, however, is frequently lacking during marine reserve design and establishment. We characterise patterns of genetic connectivity of 3 key species of habitat-forming macroalgae across an established network of temperate marine reserves on the east coast of Australia and the implications for adaptive management and marine reserve design. Connectivity varied greatly among species. Connectivity was high for the subtidal macroalgae Ecklonia radiata and Phyllospora comosa and neither species showed any clear patterns of genetic structuring with geographic distance within or among marine parks. In contrast, connectivity was low for the intertidal, Hormosira banksii, and there was a strong pattern of isolation by distance. Coastal topography and latitude influenced small scale patterns of genetic structure. These results suggest that some species are well served by the current system of marine reserves in place along this temperate coast but it may be warranted to revisit protection of intertidal habitats to ensure the long-term persistence of important habitat-forming macroalgae. Adaptively managing marine reserve design to maintain connectivity may ensure the long-term persistence and resilience of marine habitats and the biodiversity they support. PMID- 21625389 TI - Own song selectivity in the songbird auditory pathway: suppression by norepinephrine. AB - BACKGROUND: Like human speech, birdsong is a learned behavior that supports species and individual recognition. Norepinephrine is a catecholamine suspected to play a role in song learning. The goal of this study was to investigate the role of norepinephrine in bird's own song selectivity, a property thought to be important for auditory feedback processes required for song learning and maintenance. METHODOLOGY/PRINCIPAL FINDINGS: Using functional magnetic resonance imaging, we show that injection of DSP-4, a specific noradrenergic toxin, unmasks own song selectivity in the dorsal part of NCM, a secondary auditory region. CONCLUSIONS/SIGNIFICANCE: The level of norepinephrine throughout the telencephalon is known to be high in alert birds and low in sleeping birds. Our results suggest that norepinephrine activity can be further decreased, giving rise to a strong own song selective signal in dorsal NCM. This latent own song selective signal, which is only revealed under conditions of very low noradrenergic activity, might play a role in the auditory feedback and/or the integration of this feedback with the motor circuitry for vocal learning and maintenance. PMID- 21625390 TI - Interleukin-22 suppresses the growth of A498 renal cell carcinoma cells via regulation of STAT1 pathway. AB - BACKGROUND: Renal cell carcinoma (RCC) is one of the most common kidney cancers and is highly resistant to chemotherapy. Accumulating evidence suggests that interleukin-22 (IL-22) may mediate host defense against varietal pathogens as a proinflammatory and anti-inflammatory cytokine. The purpose of this study is to assess the inhibitory effects of IL-22 on human RCC cell line A498 and to investigate the possible mechanisms underlying the anti-tumor effects of this cytokine. METHODOLOGY: A498 cells, a RCC cell line, were used to assess the inhibitory growth effects of IL-22 using the MTT assay and flow cytometric analysis in vitro. BALB/C nude mice bearing A498 cell xenografts were used to examine the antitumor efficacy of IL-22 in vivo. Western blotting assay was performed to detect the regulation of the intracellular signaling pathway of IL 22. PRINCIPAL FINDINGS: We found that IL-22 suppressed the growth of A498 cells in a dose-dependent manner and inhibited the growth of A498 xenografts. We also observed that IL-22 produced a dose-dependent inhibitory effect on A498 cells that involved the induction of G2/M cell cycle arrest without cell apoptosis. Moreover, we showed that the phosphorylation of STAT1 was increased and the phosphorylation of ERK1/2 was attenuated in A498 cells exposed to IL-22. The growth inhibition of A498 cells was partially revised after IL-22 treatment as the expression of STAT1 was knocked down. And inflammatory cytokines, interferon alpha and tumor necrosis factor-alpha (TNF-alpha) were barely involved in the suppression of A498 cell xenografts treated with IL-22. CONCLUSIONS: IL-22 dose dependently suppresses RCC cell line A498 cells in vitro and induces growth inhibition of A498 cell-bearing mouse xenografts. These results suggest that the anti-RCC effects of IL-22 are at least partially mediated through regulation of STAT1 signaling pathways and G2/M cell cycle arrest, rather than by inducing apoptosis and inflammatory cytokines. PMID- 21625391 TI - Functional characterization of a lipoprotein-encoding operon in Campylobacter jejuni. AB - BACKGROUND: Bacterial lipoproteins have important functions in bacterial pathogenesis and physiology. In Campylobacter jejuni, a major foodborne pathogen causing gastroenteritis in humans, the majority of lipoproteins have not been functionally characterized. Previously, we showed by DNA microarray that CmeR, a transcriptional regulator repressing the expression of the multidrug efflux pump CmeABC, modulates the expression of a three-gene operon (cj0089, cj0090, and cj0091) encoding a cluster of lipoproteins in C. jejuni. METHODOLOGY/PRINCIPAL FINDINGS: In this work, we characterized the function and regulation of the cj0089-cj0090-cj0091 operon. In contrast to the repression of cmeABC, CmeR activates the expression of the lipoprotein genes and the regulation is confirmed by immunoblotting using anti-Cj0089 and anti-Cj0091 antibodies. Gel mobility shift assay showed that CmeR directly binds to the promoter of the lipoprotein operon, but the binding is much weaker compared with the promoter of cmeABC. Analysis of different cellular fractions indicated that Cj0089 was associated with the inner membrane, while Cj0091 was located on the outer membrane. Inactivation of cj0091, but not cj0089, significantly reduced the adherence of C. jejuni to INT 407 cells in vitro, indicating that Cj0091 has a function in adherence. When inoculated into chickens, the Cj0091 mutant also showed a defect in early colonization of the intestinal tract, suggesting that Cj0091 contributes to Campylobacter colonization in vivo. It was also shown that Cj0091 was produced and immunogenic in chickens that were naturally infected by C. jejuni. CONCLUSION/SIGNIFICANCE: These results indicate that the lipoprotein operon is subject to direct regulation by CmeR and that Cj0091 functions as an adhesion mechanism in C. jejuni and contributes to Campylobacter colonization of the intestinal tract in animal hosts. PMID- 21625392 TI - Evidence-Based Strategies Improve Assessment of Pediatric Bipolar Disorder by Community Practitioners. AB - The misdiagnosis of pediatric bipolar disorder (PBD) has become a major public health concern. Would available evidence-based assessment (EBA) strategies help improve diagnostic accuracy, and are clinicians willing to consider these strategies in practice? The purpose of the present study was to document the extent to which using an EBA decision tool--a probability nomogram--improves the interpretation of family history and test data by clinicians and to examine the acceptability of the nomogram technique to clinicians. Over 600 clinicians across the US and Canada attending continuing education seminars were trained to use the nomogram. Participants estimated the probability that a youth in a clinical vignette had bipolar disorder, first using clinical judgment and then using the nomogram. Brief training of clinicians (less than 30 minutes) in using the nomogram for assessing PBD improved diagnostic accuracy, consistency, and agreement. The majority of clinicians endorsed using the nomogram in practice. EBA decision aids, such as the nomogram, may lead to a significant decrease in overdiagnosis and help clinicians detect true cases of PBD. PMID- 21625393 TI - Iso-migrastatin Titer Improvement in the Engineered Streptomyces lividans SB11002 Strain by Optimization of Fermentation Conditions. AB - The heterologous production of iso-migrastatin (iso-MGS) was successfully demonstrated in an engineered S. lividans SB11002 strain, which was derived from S. lividans K4-114, following introduction of pBS11001, which harbored the entire mgs biosynthetic gene cluster. However, under similar fermentation conditions, the iso-MGS titer in the engineered strain was significantly lower than that in the native producer - Streptomyces platensis NRRL 18993. To circumvent the problem of low iso-MGS titers and to expand the utility of this heterologous system for iso-MGS biosynthesis and engineering, systematic optimization of the fermentation medium was carried out. The effects of major components in the cultivation medium, including carbon, organic and inorganic nitrogen sources, were investigated using a single factor optimization method. As a result, sucrose and yeast extract were determined to be the best carbon and organic nitrogen sources, resulting in optimized iso-MGS production. Conversely, all other inorganic nitrogen sources evaluated produced various levels of inhibition of iso MGS production. The final optimized R2YE production medium produced iso-MGS with a titer of 86.5 mg/L, about 3.6-fold higher than that in the original R2YE medium, and 1.5 fold higher than that found within the native S. platensis NRRL 18993 producer. PMID- 21625394 TI - Removal of Trace Arsenic to Meet Drinking Water Standards Using Iron Oxide Coated Multiwall Carbon Nanotubes. AB - This study presents the removal of trace level arsenic to meet drinking water standards using an iron oxide-multi-walled carbon nanotube (Fe-MWCNT) hybrid as a sorbent. The synthesis was facilitated by the high degree of nanotube functionalization using a microwave assisted process, and a controlled assembly of iron oxide was possible where the MWCNT served as an effective support for the oxide. In the final product, 11 % of the carbon atoms were attached to Fe. The Fe MWCNT was effective in arsenic removal to below the drinking water standard levels of 10 ug L(-1). The absorption capacity of the composite was 1723 ug g(-1) and 189 ug g(-1) for As(III) and As(V) respectively. The adsorption of As(V) on Fe-MWCNT was faster than that of As(III). The pseudo-second order rate equation was found to effectively describe the kinetics of arsenic adsorption. The adsorption isotherms for As(III) and As(V) fitted both the Langmuir and Freundlich models. PMID- 21625395 TI - Patient-Specific Geometry Modeling and Mesh Generation for Simulating Obstructive Sleep Apnea Syndrome Cases by Maxillomandibular Advancement. AB - The objective of this paper is the reconstruction of upper airway geometric models as hybrid meshes from clinically used Computed Tomography (CT) data sets in order to understand the dynamics and behaviors of the pre- and postoperative upper airway systems of Obstructive Sleep Apnea Syndrome (OSAS) patients by viscous Computational Fluid Dynamics (CFD) simulations. The selection criteria for OSAS cases studied are discussed because two reasonable pre- and postoperative upper airway models for CFD simulations may not be created for every case without a special protocol for CT scanning. The geometry extraction and manipulation methods are presented with technical barriers that must be overcome so that they can be used along with computational simulation software as a daily clinical evaluation tool. Eight cases are presented in this paper, and each case consists of pre- and postoperative configurations. The results of computational simulations of two cases are included in this paper as demonstration. PMID- 21625396 TI - Characterization of PM(2.5) collected during broadcast and slash-pile prescribed burns of predominately ponderosa pine forests in northern Arizona. AB - Prescribed burning, in combination with mechanical thinning, is a successful method for reducing heavy fuel loads from forest floors and thereby lowering the risk of catastrophic wildfire. However, an undesirable consequence of managed fire is the production of fine particulate matter or PM(2.5) (particles <=2.5 um in aerodynamic diameter). Wood-smoke particulate data from 21 prescribed burns are described, including results from broadcast and slash-pile burns. All PM(2.5) samples were collected in situ on day 1 (ignition) or day 2. Samples were analyzed for mass, polycyclic aromatic hydrocarbons (PAHs), inorganic elements, organic carbon (OC), and elemental carbon (EC). Results were characteristic of low intensity, smoldering fires. PM(2.5) concentrations varied from 523 to 8357 ug m(-3) and were higher on day 1. PAH weight percents (19 PAHs) were higher in slash-pile burns (0.21 +/- 0.08% OC) than broadcast burns (0.07 +/- 0.03% OC). The major elements were K, Cl, S, and Si. OC and EC values averaged 66 +/- 7 and 2.8 +/- 1.4% PM(2.5), respectively, for all burns studied, in good agreement with literature values for smoldering fires. PMID- 21625398 TI - Intercellular cancer collisions generate an ejected crystal comet tail effect with fractal interface embryoid body reassembly transformation. AB - We have documented self-assembled geometric triangular chiral crystal complexes (GTCHC) and a framework of collagen vascular invariant geometric attractors in cancer tissues. This article shows how this system evolves in time. These structures are incorporated together and evolve in different ways. When the geometric core is stable, and the tissue architecture collapses, fragmented components emerge, which reveal a hidden interior identifying how each molecule is reassembled into the original mold, using one common connection, ie, a fractal self-similarity that guided the system from the beginning. GTCHC complexes generate ejected crystal comet tail effects and produce strange helicity states that arise in the form of spin domain interactions. As the crystal growth vibration stage progresses, biofractal echo images converge in a master-built construction of embryoid bodies with enolase-selective immunopositivity in relation to clusters of triangular chiral cell organization. In our electro-optic collision model, we were able to predict and replicate all the characteristics of this complex geometry that connects a physical phenomenon with the signal patterns that generate biologic chaos. Intrinsically, fractal geometry makes spatial correction errors embrace the chaotic system in a way that permits new structures to emerge, and as a result, an ordered self-assembly of embryoid bodies with neural differentiation at the final stage of cancer development is a predictable process. We hope that further investigation of these structures will lead not only to a new way of thinking about physics and biology, but also to a rewarding area in cancer research. PMID- 21625397 TI - Interpreting clinical assays for histone deacetylase inhibitors. AB - As opposed to genetics, dealing with gene expressions by direct DNA sequence modifications, the term epigenetics applies to all the external influences that target the chromatin structure of cells with impact on gene expression unrelated to the sequence coding of DNA itself. In normal cells, epigenetics modulates gene expression through all development steps. When "imprinted" early by the environment, epigenetic changes influence the organism at an early stage and can be transmitted to the progeny. Together with DNA sequence alterations, DNA aberrant cytosine methylation and microRNA deregulation, epigenetic modifications participate in the malignant transformation of cells. Their reversible nature has led to the emergence of the promising field of epigenetic therapy. The efforts made to inhibit in particular the epigenetic enzyme family called histone deacetylases (HDACs) are described. HDAC inhibitors (HDACi) have been proposed as a viable clinical therapeutic approach for the treatment of leukemia and solid tumors, but also to a lesser degree for noncancerous diseases. Three epigenetic drugs are already arriving at the patient's bedside, and more than 100 clinical assays for HDACi are registered on the National Cancer Institute website. They explore the eventual additive benefits of combined therapies. In the context of the pleiotropic effects of HDAC isoforms, more specific HDACi and more informative screening tests are being developed for the benefit of the patients. PMID- 21625399 TI - Dose study of the multikinase inhibitor, LY2457546, in patients with relapsed acute myeloid leukemia to assess safety, pharmacokinetics, and pharmacodynamics. AB - BACKGROUND: Acute myeloid leukemia (AML) is a life-threatening malignancy with limited treatment options in chemotherapy-refractory patients. A first-in-human dose study was designed to investigate a safe and biologically effective dose range for LY2457546, a novel multikinase inhibitor, in patients with relapsed AML. METHODS: In this nonrandomized, open-label, dose escalation Phase I study, LY2457546 was administered orally once a day. Safety, pharmacokinetics, changes in phosphorylation of target kinases in AML blasts, and risk of drug-drug interactions (DDI) were assessed. RESULTS: Five patients were treated at the starting and predicted minimal biologically effective dose of 50 mg/day. The most commonly observed adverse events were febrile neutropenia, epistaxis, petechiae, and headache. The majority of adverse events (81%) were Grade 1 or 2. One patient had generalized muscle weakness (Grade 3), which was deemed to be a dose-limiting toxicity. Notably, the pharmacokinetic profile of LY2457546 showed virtually no elimination of LY2457546 within 24 hours, and thus prevented further dose escalation. No significant DDI were observed. Ex vivo flow cytometry studies showed downregulation of the phosphoproteins, pcKIT, pFLT3, and pS6, in AML blasts after LY2457546 administration. No medically relevant responses were observed in the five treated patients. CONCLUSION: No biologically effective dose could be established for LY2457546 in chemotherapy-resistant AML patients. Lack of drug clearance prevented safe dose escalation, and the study was terminated early. Future efforts should be made to develop derivatives with a more favorable pharmacokinetic profile. PMID- 21625400 TI - Clinical significance of interleukin (IL)-6 in cancer metastasis to bone: potential of anti-IL-6 therapies. AB - Metastatic events to the bone occur frequently in numerous cancer types such as breast, prostate, lung, and renal carcinomas, melanoma, neuroblastoma, and multiple myeloma. Accumulating evidence suggests that the inflammatory cytokine interleukin (IL)-6 is frequently upregulated and is implicated in the ability of cancer cells to metastasize to bone. IL-6 is able to activate various cell signaling cascades that include the STAT (signal transducer and activator of transcription) pathway, the PI3K (phosphatidylinositol-3 kinase) pathway, and the MAPK (mitogen-activated protein kinase) pathway. Activation of these pathways may explain the ability of IL-6 to mediate various aspects of normal and pathogenic bone remodeling, inflammation, cell survival, proliferation, and pro-tumorigenic effects. This review article will discuss the role of IL-6: 1) in bone metabolism, 2) in cancer metastasis to bone, 3) in cancer prognosis, and 4) as potential therapies for metastatic bone cancer. PMID- 21625401 TI - Real Time FRET Based Detection of Mechanical Stress in Cytoskeletal and Extracellular Matrix Proteins. AB - A molecular force sensing cassette (stFRET) was incorporated into actinin, filamin, and spectrin in vascular endothelial cells (BAECs) and into collagen-19 in Caenorhabditis elegans. To estimate the stress sensitivity of stFRET in solution, we used DNA springs. A 60-mer loop of single stranded DNA was covalently linked to the external cysteines of the donor and acceptor. When the complementary DNA was added it formed double stranded DNA with higher persistence length, stretching the linker and substantially reducing FRET efficiency. The probe stFRET detected constitutive stress in all cytoskeletal proteins tested, and in migrating cells the stress was greater at the leading edge than the trailing edge. The stress in actinin, filamin and spectrin could be reduced by releasing focal attachments from the substrate with trypsin. Inhibitors of actin polymerization produced a modest increase in stress on the three proteins suggesting they are mechanically in parallel. Local shear stress applied to the cell with a perfusion pipette showed gradients of stress leading from the site of perfusion. Transgenic C. elegans labeled in collagen-19 produced a behaviorally and anatomically normal animal with constitutive stress in the cuticle. Stretching the worm visibly stretched the probe in collagen showing that we can trace the distribution of mean tissue stress in specific molecules. stFRET is a general purpose dynamic sensor of mechanical stress that can be expressed intracellularly and extracellularly in isolated proteins, cells, tissues, organs and animals. PMID- 21625402 TI - Composition and consistency of the desired affective state: The role of personality and motivation. AB - Using longitudinal and experience sampling designs, the consistency and composition, and personality and motivational predictors, of the desired affective state are explored. Findings indicate that, while the desired affect is relatively malleable throughout one semester, it is relatively stable throughout 1 week. Personality and motivations/goals were related to the content of the desired affective state. Extraversion, neuroticism, agreeableness, and conscientiousness were related to the content of the desired affective state. In addition, higher-order goals predicted the content of the desired affective state. Our results suggest that the content of the desired affective state may be largely dependent on personality, motivation, and, potentially, an interaction between personality and motivation. PMID- 21625403 TI - A Qualitative Assessment of South African Adolescents' Motivations For and Against Substance Use and Sexual Behavior. AB - Focus groups (N = 15 groups; eight with girls, seven with boys) with adolescents in high schools near Cape Town, South Africa were used to conduct a qualitative investigation of reported reasons for using and not using substances, and for having and not having sex. Adolescents reported Enhancement, Negative States, Social, and Aversive Social motivations for both substance use and sexual behavior. In addition, being addicted as a reason for using drugs and rape as a context for sexual behavior were frequently reported. Motivations against behaviors included Physical/Behavioral Consequences, Ethical Objections, Social Disapproval, and Activities or Future Orientation reasons. Preventive interventions should address existing motivations for and against substance use and sexual behavior to acknowledge adolescents' experiences in context. PMID- 21625404 TI - Beyond Fast Mapping. AB - Since the seminal 1957 studies of word learning by Roger Brown, most experimental studies of lexical acquisition have concerned fast mapping: the process through which a new lexical entry is established, and through which representations of the linguistic context of a newly heard word interact with representations of its nonlinguistic context to fix an initial partial meaning. Here I focus on the subsequent extended process through which the adult meaning is approximated. Two factors lead to an extended learning process; the size of the hypothesis space and the need, sometimes, for the creation of new semantic primitives. Sometimes lexical learning requires conceptual change. I sketch a learning mechanism through which this can be achieved. A case study of learning the meanings of verbal numerals illustrates the argument. PMID- 21625405 TI - The Effects of Source Credibility in the Presence or Absence of Prior Attitudes: Implications for the Design of Persuasive Communication Campaigns. AB - Most theories of persuasion predict that limited ability and motivation to think about communications should increase the impact of source credibility on persuasion. Furthermore, this effect is assumed to occur, regardless of whether or not the recipients have prior attitudes. In this study, the effects of source credibility, ability, and motivation (knowledge, message repetition, relevance) on persuasion were examined meta-analytically across both attitude formation and change conditions. Findings revealed that the Source Credibility * Ability/Motivation interaction emerged only when participants lacked prior attitudes and were unable to form a new attitude based on the message content. In such settings, the effects of source credibility decayed rapidly. The implications of these findings for applied communication campaigns are discussed. PMID- 21625406 TI - Designing Neural Networks in Culture: Experiments are described for controlled growth, of nerve cells taken from rats, in predesigned geometrical patterns on laboratory culture dishes. AB - Technology has advanced to where it is possible to design and grow-with predefined geometry and surprisingly good fidelity-living networks of neurons in culture dishes. Here we overview the elements of design, emphasizing the lithographic techniques that alter the cell culture surface which in turn influences the attachment and growth of the neural networks. Advanced capability in this area makes it possible to design networks of desired complexity. Other issues addressed include the influence of glial cells and media on activity and the potential for extending the designs into three dimensions. Investigators are advancing the art and science of analyzing and controlling through stimulation the function of the neural networks, including the ability to take advantage of their geometric form in order to influence functional properties. PMID- 21625407 TI - Complex home care: Part I--Utilization and costs to families for health care services each year. AB - As many as 120 persons per million people in the United States are dependent on the lifelong, complex, technology-based care of home parenteral nutrition (HPN) infusions. However, data for costs paid by families for HPN-related health care services and for non-reimbursed expenditures are rarely tabulated and most often underestimated. The goals of this study were to describe health care services used by families to manage HPN, report the frequency of each service used annually, and estimate the average annual non-reimbursed costs to families for these health services. The numerous and varied types of services reported and the time required to coordinate and access HPN services illustrates the challenges faced by patients and their family caregivers. The lack of a coordinated and efficient system for delivering complex chronic care results in poorer outcomes for HPN patients and their families on-reimbursed costs and the extensive amount of time required to coordinate multi-professional services negatively impacts the clinical outcomes and quality of life of complex chronic home care. PMID- 21625408 TI - Pb-207 NMR spectroscopy reveals that Pb(II) coordinates with glutathione (GSH) and tris cysteine zinc finger proteins in a PbS3 coordination environment. AB - 207Pb NMR spectroscopy can be used to monitor the binding of Pb(II) to thiol rich biological small molecules such as glutathione and to zinc finger proteins. The UV/visible (UV/Vis) absorption band centered at 334 nM and the observed 207Pb signal in 207Pb NMR (delta~5750 ppm) indicate that glutathione binds Pb(II) in a trigonal pyramidal geometry (PbS3) at pH 7.5 or higher with a 1:3 molar ratio of Pb(II) to GSH. While previous studies using UV/Vis and extended X-ray absorption fine structure (EXAFS) spectroscopy were interpreted to show that the zinc binding domain from HIV nucleocapsid protein (HIV-CCHC) binds Pb(II) in a single PbS3 environment, the more sensitive 207Pb NMR spectra (at pH 7.0, 1:1 molar ratio) provide compelling evidence for the presence of two PbS3 structures (delta=5790 and 5744 ppm), one of which is more stable at high temperatures. It has previously been proposed that the HIV-CCHH peptide does not fold properly to afford a PbS2N motif, because histidine does not bind to Pb(II). These predictions are confirmed by the present studies. These results demonstrate the applicability of 207Pb NMR to biomolecular structure determination in proteins with cysteine binding sites for the first time. PMID- 21625409 TI - Modular Ontology Techniques and their Applications in the Biomedical Domain. AB - In the past several years, various ontologies and terminologies such as the Gene Ontology have been developed to enable interoperability across multiple diverse medical information systems. They provide a standard way of representing terms and concepts thereby supporting easy transmission and interpretation of data for various applications. However, with their growing utilization, not only has the number of available ontologies increased considerably, but they are also becoming larger and more complex to manage. Toward this end, a growing body of work is emerging in the area of modular ontologies where the emphasis is on either extracting and managing "modules" of an ontology relevant to a particular application scenario (ontology decomposition) or developing them independently and integrating into a larger ontology (ontology composition). In this paper, we investigate state-of-the-art approaches in modular ontologies focusing on techniques that are based on rigorous logical formalisms as well as well-studied graph theories. We analyze and compare how such approaches can be leveraged in developing tools and applications in the biomedical domain. We conclude by highlighting some of the limitations of the modular ontology formalisms and put forward additional requirements to steer their future development. PMID- 21625410 TI - Risk Factors of Early and Late Onset Preeclampsia among Thai Women. AB - BACKGROUND: Little research has been conducted to specifically identify risk factors of early and late onset preeclampsia among Thai women. OBJECTIVE: To examine risk factors of early and late onset of preeclampsia among Thai women. METHODS: A case-control study of 150 preeclampsia cases with an equal number of normotensive controls was conducted among women who delivered live born singleton infants at King Chulalongkorn Memorial Hospital, Rajavithi Hospital, and Police General Hospital in Bangkok, Thailand from July 2006 to November 2007. Multivariable logistic regression analysis procedures were used to calculate odds ratios (OR) and 95% confidence intervals (CI) of potential risk factors associated with preeclampsia. RESULTS: Pre-pregnancy body mass index >30 kg/m2 (OR=5.25, 95%CI: 1.80, 15.32) and failure to use prenatal care services (OR=6.37, 95%CI: 1.26, 32.27) were associated with increased risk of preeclampsia. OR's of similar magnitude were observed when risk factors of early and late onset preeclampsia were assessed separately. CONCLUSION: Advanced maternal age, obesity, and no utilization of prenatal care were covariates identified as risk factors for preeclampsia. PMID- 21625411 TI - The impact of food price increases on caloric intake in China. AB - World food prices have increased dramatically in recent years. We use panel data from 2006 to examine the impact of these increases on the consumption and nutrition of poor households in two Chinese provinces. We find that households in Hunan suffered no nutrition declines. Households in Gansu experienced a small decline in calories, though the decline is on par with usual seasonal effects. The overall nutritional impact of the world price increase was small because households were able to substitute to cheaper foods and because the domestic prices of staple foods remained low due to government intervention in grain markets. PMID- 21625412 TI - Adopting Graph Traversal Techniques for Context-Driven Value Sets Extraction from Biomedical Knowledge Sources. AB - The ability to model, share and re-use value sets across multiple medical information systems is an important requirement. However, generating value sets semi-automatically from a terminology service is still an unresolved issue, in part due to the lack of linkage to clinical context patterns that provide the constraints in defining a concept domain and invocation of value sets extraction. Towards this goal, we develop and evaluate an approach for context-driven automatic value sets extraction based on a formal terminology model. The crux of the technique is to identify and define the context patterns from various domains of discourse and leverage them for value set extraction using two complementary ideas based on (i) local terms provided by the subject matter experts (extensional) and (ii) semantic definition of the concepts in coding schemes (intensional). We develop algorithms based on well-studied graph traversal and ontology segmentation techniques for both the approaches and implement a prototype demonstrating their applicability on use cases from, SNOMED CT rendered, in the LexGrid terminology model. We also present preliminary evaluation of our approach and report investigation results done by subject matter experts at the Mayo Clinic. PMID- 21625413 TI - Association of Godoy & Godoy contention with mechanism with apparatus-assisted exercises in patients with arm lymphedema after breast cancer. AB - AIM: The aim of the current study was to evaluate the reduction in the volume of the upper limbs with lymphedema after exercises using the apparatus-assisted program associated with contention mechanism. PATIENTS AND METHOD: Twenty-eight female patients were selected and referred for evaluation and treatment of lymphedema after breast cancer therapy. The ages of the women ranged from 42 to 72 years with a mean age of 57 years. Inclusion criteria were treatment of cancer associated to a difference of at least 200 mL between the edematous and the contralateral limbs. Patients with active infections, skin lesions, and active disease were not included in the study. Four series of exercises using devices based on pedals, pulleys, a horizontal reflexion bar, and an elevation bar were selected. The participants were advised about the form of exercise: 15 minutes for each device, low intensity (less than 10 movements per minute), in the seated position, and the use of contention. Water displacement volumetry was performed before and after the 60-minute exercise session. The paired t-test was utilized with an alpha error of 5% considered acceptable (P value < 0.05). RESULTS: The mean difference between the volumetric measures before and after exercise was significant, with all the participants having reductions in the volume of the limbs using the four selected devices over time and at an intensity determined by this study. CONCLUSION: Association of a Godoy and Godoy contention during apparatus-assisted exercise reduced the edema in patients with lymphedema of the upper limbs. PMID- 21625414 TI - Diagnosis of depression by MRI scans with the use of VSRAD - a promising auxiliary means of diagnosis: a report of 10 years research. AB - OBJECTIVES: This study was designed to evaluate the usefulness of assessing subgenual anterior cingulate cortex (sACC) volume reduction by magnetic resonance imaging (MRI) as an objective auxiliary means of diagnosis of depression. The study was additionally designed to analyze the association of sACC volume reduction with the effectiveness of treatments for depression and other diseases presenting with similar symptoms, and to examine the possibility of using sACC volume reduction in the distinction between depression and bipolar disorder and determining optimum medication for these conditions. METHODS: Three-dimensional T1-weighted sagittal images, taken with Achieva 1.5T NOVA (Philips), were analyzed with VSRAD plus((r)) to evaluate a reduction in sACC volume. The finding from this analysis was compared with the clinical data, including the longitudinal course follow-up data based on the treatment algorithm. SUBJECTS: The study involved 88 patients aged over 54 who received MRI during 2010, ie, 71 patients with major depressive disorder (MDD), 11 patients bipolar disorder, and 6 patients in whom the initial diagnosis (MDD) was later modified. Thirty-three normal individuals served as controls. RESULTS: sACC volume reduction was noted in 66 of the 71 patients receiving treatment of MDD, with sensitivity of 93%, specificity of 85%, and accuracy of 90%. In the 66 patients diagnosed as having MDD and exhibiting sACC volume reduction, the disease showed remission in response to treatment with antidepressants, but medication needed to be continued after achievement of remission. In cases initially diagnosed as having MDD but not exhibiting sACC volume reduction, the necessity of modifying the diagnosis was considered. Typical cases of bipolar disorder did not exhibit sACC volume reduction. CONCLUSION: That patients receiving treatment of MDD often showed reduction in sACC volume suggests the usefulness of this parameter as an objective auxiliary means of diagnosis for MDD. PMID- 21625415 TI - Dendritic cell populations in patients with self-reported food hypersensitivity. AB - Self-reported hypersensitivity to food is a common condition and many of these patients have indications of intestinal immune activation. Dendritic cells (DCs) are recognized as the most potent antigen-presenting cells involved in both initiating immune responses and maintaining tolerance. The aims of this study were to evaluate the DC populations with their phenotype and T cell stimulatory capacity in patients with food hypersensitivity and to study its relationship with atopic disease. Blood samples from 10 patients with self-reported food hypersensitivity, divided into atopic and nonatopic subgroups, and 10 gender- and age-matched healthy controls were analyzed by flow cytometry using the Miltenyi Blood Dendritic cells kit. Monocyte-derived DCs (moDCs) were evaluated concerning their phenotype and T cell stimulatory capacity. DC populations and cell surface markers were not significantly different between patients and healthy controls, but moDCs from atopic patients expressed significantly more CD38 compared to moDCs from nonatopic patients. Moreover, lipopolysaccharide stimulated moDCs from atopic patients produced significantly more interleukin-10 compared to nonatopic patients. CD38 expression was correlated to total serum immunoglobulin E levels. These findings support the notion of immune activation in some patients with self reported food hypersensitivity. They need to be confirmed in a larger cohort. PMID- 21625416 TI - Development of oral immunomodulatory agents in the management of multiple sclerosis. AB - The emergence of oral disease-modifying therapies in multiple sclerosis (MS) will have a significant impact on the evolving scenario of immunomodulatory treatments in MS where current therapies are all injectable. Reducing relapses in trials translates for individuals with MS into a therapeutic aim of stopping future events. Thus the possible absence of any perceived benefits to the individual together with the long disease course, variable outcome, and a younger age group affected in MS makes side effects the major issue. The use of disease-modifying therapies as a whole needs to be placed in the context of a widening therapeutic indication where the use of these therapies is being justified at an increasingly early stage and in pre-MS syndromes such as clinically isolated and radiologically isolated syndromes where no fixed disability is likely to have accumulated. The five oral therapies discussed (cladribine, fingolimod, laquinimod, BG-12, and teriflunomide) have just completed Phase III studies and some have just been licensed. New oral drugs for MS need to be placed within this evolving marketplace where ease of delivery together with efficacy and side effects needs to be balanced against the known issues but also the known long term safety of standard injectables. PMID- 21625417 TI - Current difficulties and recent advances in bypass therapy for the management of hemophilia with inhibitors: a new and practical formulation of recombinant factor VIIa. AB - Bypassing agents are the mainstay of treatment for patients with hemophilia with high-titer inhibitors. Whereas the availability of these agents has greatly advanced the management of bleeding episodes in this population, timely administration of bypassing agents continues to be hampered by a number of practical limitations, including the need for refrigerated storage of the agent and its reconstitution at room temperature prior to administration, among others. In this review, the importance of early treatment of bleeds and factors that influence this more timely therapeutic approach are highlighted, together with the advantages offered by the use of a new formulation of recombinant activated factor VII that permits improved storage and portability, potentially optimizing timely bypassing agent administration. PMID- 21625418 TI - Place of pitavastatin in the statin armamentarium: promising evidence for a role in diabetes mellitus. AB - Inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, known as statins, have revolutionized the treatment of hypercholesterolemia and coronary artery disease prevention. However, there are considerable issues regarding statin safety and further development of residual risk control, particularly for diabetic and metabolic syndrome patients. Pitavastatin is a potent statin with low-density lipoprotein (LDL) cholesterol-lowering effects comparable to those of atorvastatin or rosuvastatin. Pitavastatin has a high-density lipoprotein (HDL) cholesterol raising effect, may improve insulin resistance, and has little influence on glucose metabolism. Considering these factors along with its unique pharmacokinetic properties, which suggest minimal drug-drug interaction, pitavastatin could provide an alternative treatment choice, especially in patients with glucose intolerance or diabetes mellitus. Many clinical trials are now underway to test the clinical efficacy of pitavastatin in various settings and are expected to provide further information. PMID- 21625419 TI - Flavonoids targeting of IkappaB phosphorylation abrogates carcinogen-induced MMP 9 and COX-2 expression in human brain endothelial cells. AB - Brain endothelial cells play an essential role as structural and functional components of the blood-brain barrier (BBB). Increased BBB breakdown and brain injury are associated with neuroinflammation and are thought to trigger mechanisms involving matrix metalloproteinase upregulation. Emerging evidence also indicates that cyclooxygenase (COX) inhibition limits BBB disruption, but the mechanisms linking metalloproteinase to COX remain unknown. In this study, we sought to investigate the nuclear factor-kappa B (NF-kappaB) signaling pathway, a common pathway in both the regulation of matrix metalloproteinase-9 (MMP-9) and COX-2 expression, and the inhibitory properties of several chemopreventive flavonoids. Human brain microvascular endothelial cells were treated with a combination of phorbol 12-myristate 13-acetate (PMA), a carcinogen documented to increase MMP-9 and COX-2 through NF-kappaB, and several naturally occurring flavonoids. Among the molecules tested, we found that fisetin, apigenin, and luteolin specifically and dose-dependently antagonized PMA-induced COX-2 and MMP 9 gene and protein expressions as assessed by qRT-PCR, immunoblotting, and zymography respectively. We further demonstrate that flavonoids impact on IkappaK mediated phosphorylation activity as demonstrated by the inhibition of PMA induced IkappaB phosphorylation levels. Our results suggest that BBB disruption during neuroinflammation could be pharmacologically reduced by a specific class of flavonoids acting as NF-kappaB signal transduction inhibitors. PMID- 21625420 TI - PDlim2 selectively interacts with the PDZ binding motif of highly pathogenic avian H5N1 influenza A virus NS1. AB - The multi-functional NS1 protein of influenza A virus is a viral virulence determining factor. The last four residues at the C-terminus of NS1 constitute a type I PDZ domain binding motif (PBM). Avian influenza viruses currently in circulation carry an NS1 PBM with consensus sequence ESEV, whereas human influenza viruses bear an NS1 PBM with consensus sequence RSKV or RSEV. The PBM sequence of the influenza A virus NS1 is reported to contribute to high viral pathogenicity in animal studies. Here, we report the identification of PDlim2 as a novel binding target of the highly pathogenic avian influenza virus H5N1 strain with an NS1 PBM of ESEV (A/Chicken/Henan/12/2004/H5N1, HN12-NS1) by yeast two hybrid screening. The interaction was confirmed by in vitro GST pull-down assays, as well as by in vivo mammalian two-hybrid assays and bimolecular fluorescence complementation assays. The binding was also confirmed to be mediated by the interaction of the PDlim2 PDZ domain with the NS1 PBM motif. Interestingly, our assays showed that PDlim2 bound specifically with HN12-NS1, but exhibited no binding to NS1 from a human influenza H1N1 virus bearing an RSEV PBM (A/Puerto Rico/8/34/H1N1, PR8-NS1). A crystal structure of the PDlim2 PDZ domain fused with the C-terminal hexapeptide from HN12-NS1, together with GST pull-down assays on PDlim2 mutants, reveals that residues Arg16 and Lys31 of PDlim2 are critical for the binding between PDlim2 and HN12-NS1. The identification of a selective binding target of HN12-NS1 (ESEV), but not PR8-NS1 (RSEV), enables us to propose a structural mechanism for the interaction between NS1 PBM and PDlim2 or other PDZ-containing proteins. PMID- 21625421 TI - Biomarkers in natural fish populations indicate adverse biological effects of offshore oil production. AB - BACKGROUND: Despite the growing awareness of the necessity of a sustainable development, the global economy continues to depend largely on the consumption of non-renewable energy resources. One such energy resource is fossil oil extracted from the seabed at offshore oil platforms. This type of oil production causes continuous environmental pollution from drilling waste, discharge of large amounts of produced water, and accidental spills. METHODS AND PRINCIPAL FINDINGS: Samples from natural populations of haddock (Melanogrammus aeglefinus) and Atlantic cod (Gadus morhua) in two North Sea areas with extensive oil production were investigated. Exposure to and uptake of polycyclic aromatic hydrocarbons (PAHs) were demonstrated, and biomarker analyses revealed adverse biological effects, including induction of biotransformation enzymes, oxidative stress, altered fatty acid composition, and genotoxicity. Genotoxicity was reflected by a hepatic DNA adduct pattern typical for exposure to a mixture of PAHs. Control material was collected from a North Sea area without oil production and from remote Icelandic waters. The difference between the two control areas indicates significant background pollution in the North Sea. CONCLUSION: It is most remarkable to obtain biomarker responses in natural fish populations in the open sea that are similar to the biomarker responses in fish from highly polluted areas close to a point source. Risk assessment of various threats to the marine fish populations in the North Sea, such as overfishing, global warming, and eutrophication, should also take into account the ecologically relevant impact of offshore oil production. PMID- 21625422 TI - Parkin mediates apparent E2-independent monoubiquitination in vitro and contains an intrinsic activity that catalyzes polyubiquitination. AB - BACKGROUND: Mutations in the parkin gene, which encodes a ubiquitin ligase (E3), are a major cause of autosomal recessive parkinsonism. Although parkin-mediated ubiquitination was initially linked to protein degradation, accumulating evidence suggests that the enzyme is capable of catalyzing multiple forms of ubiquitin modifications including monoubiquitination, K48- and K63-linked polyubiquitination. In this study, we sought to understand how a single enzyme could exhibit such multifunctional catalytic properties. METHODS AND FINDINGS: By means of in vitro ubiquitination assays coupled with mass spectrometry analysis, we were surprised to find that parkin is apparently capable of mediating E2 independent protein ubiquitination in vitro, an unprecedented activity exhibited by an E3 member. Interestingly, whereas full length parkin catalyzes solely monoubiquitination regardless of the presence or absence of E2, a truncated parkin mutant containing only the catalytic moiety supports both E2-independent and E2-dependent assembly of ubiquitin chains. CONCLUSIONS: Our results here suggest a complex regulation of parkin's activity and may help to explain how a single enzyme like parkin could mediate diverse forms of ubiquitination. PMID- 21625423 TI - The antioxidative effect of electro-acupuncture in a mouse model of Parkinson's disease. AB - Accumulating evidence indicates that oxidative stress plays a critical role in Parkinson's disease (PD). Our previous work has shown that 100 Hz electro acupuncture (EA) stimulation at ZUSANLI (ST36) and SANYINJIAO (SP6) protects neurons in the substantia nigra pars compacta from 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) toxicity in male C57BL/6 mice, a model of PD. In the present study we administered 100 Hz EA stimulation at the two acupoints to MPTP lesioned mice for 12 sessions starting from the day prior to the first MPTP injection. We found that in the striatum of MPTP treated mice 100 Hz EA stimulation effectively inhibited the production of hydrogen peroxide and malonaldehyde, and increased glutathione concentration and total superoxide dismutase activity through biochemical methods. However, it decreased glutathione peroxidase activity via biochemical analysis and did not affect the level of 1 methyl-4-phenylpyridinium in the striatum revealed by high performance liquid chromatography with ultraviolet detection. These data suggest that 100 Hz EA stimulation at ST36 and SP6 has antioxidative effects in the MPTP model of PD. This data, along with our previous work, indicates that 100 Hz EA stimulation at ST36 and SP6 protects the nigrostriatal system by multiple mechanisms including antioxidation and antiapoptosis, and suggests that EA stimulation is a promising therapy for treating PD. PMID- 21625424 TI - Polyene macrolide antifungal drugs trigger interleukin-1beta secretion by activating the NLRP3 inflammasome. AB - The use of antimycotic drugs in fungal infections is based on the concept that they suppress fungal growth by a direct killing effect. However, amphotericin and nystatin have been reported to also trigger interleukin-1beta (IL-1beta) secretion in monocytes but the molecular mechanism is unknown. Here we report that only the polyene macrolides amphotericin B, nystatin, and natamycin but none of the tested azole antimycotic drugs induce significant IL-1beta secretion in vitro in dendritic cells isolated from C57BL/6 mouse bone marrow. IL-1beta release depended on Toll-like receptor-mediated induction of pro-IL-1beta as well as the NLRP3 inflammasome, its adaptor ASC, and caspase-1 for enzymatic cleavage of pro-IL-1beta into its mature form. All three drugs induced potassium efflux from the cells as a known mechanism for NLRP3 activation but the P2X7 receptor was not required for this process. Natamycin-induced IL-1beta secretion also involved phagocytosis, as cathepsin activation as described for crystal-induced IL-1beta release. Together, the polyene macrolides amphotericin B, nystatin, and natamycin trigger IL-1beta secretion by causing potassium efflux from which activates the NLRP3-ASC-caspase-1. We conclude that beyond their effects on fungal growth, these antifungal drugs directly activate the host's innate immunity. PMID- 21625425 TI - Fitness trade-offs in the evolution of dihydrofolate reductase and drug resistance in Plasmodium falciparum. AB - BACKGROUND: Patterns of emerging drug resistance reflect the underlying adaptive landscapes for specific drugs. In Plasmodium falciparum, the parasite that causes the most serious form of malaria, antifolate drugs inhibit the function of essential enzymes in the folate pathway. However, a handful of mutations in the gene coding for one such enzyme, dihydrofolate reductase, confer drug resistance. Understanding how evolution proceeds from drug susceptibility to drug resistance is critical if new antifolate treatments are to have sustained usefulness. METHODOLOGY/PRINCIPAL FINDINGS: We use a transgenic yeast expression system to build on previous studies that described the adaptive landscape for the antifolate drug pyrimethamine, and we describe the most likely evolutionary trajectories for the evolution of drug resistance to the antifolate chlorcycloguanil. We find that the adaptive landscape for chlorcycloguanil is multi-peaked, not all highly resistant alleles are equally accessible by evolution, and there are both commonalities and differences in adaptive landscapes for chlorcycloguanil and pyrimethamine. CONCLUSIONS/SIGNIFICANCE: Our findings suggest that cross-resistance between drugs targeting the same enzyme reflect the fitness landscapes associated with each particular drug and the position of the genotype on both landscapes. The possible public health implications of these findings are discussed. PMID- 21625426 TI - Granulocyte-colony stimulating factor (G-CSF) in stroke patients with concomitant vascular disease--a randomized controlled trial. AB - BACKGROUND: G-CSF has been shown in animal models of stroke to promote functional and structural regeneration of the central nervous system. It thus might present a therapy to promote recovery in the chronic stage after stroke. METHODS: Here, we assessed the safety and tolerability of G-CSF in chronic stroke patients with concomitant vascular disease, and explored efficacy data. 41 patients were studied in a double-blind, randomized approach to either receive 10 days of G-CSF (10 ug/kg body weight/day), or placebo. Main inclusion criteria were an ischemic infarct >4 months prior to inclusion, and white matter hyperintensities on MRI. Primary endpoint was number of adverse events. We also explored changes in hand motor function for activities of daily living, motor and verbal learning, and finger tapping speed, over the course of the study. RESULTS: Adverse events (AEs) were more frequent in the G-CSF group, but were generally graded mild or moderate and from the known side-effect spectrum of G-CSF. Leukocyte count rose after day 2 of G-CSF dosing, reached a maximum on day 8 (mean 42/nl), and returned to baseline 1 week after treatment cessation. No significant effect of treatment was detected for the primary efficacy endpoint, the test of hand motor function. CONCLUSIONS: These results demonstrate the feasibility, safety and reasonable tolerability of subcutaneous G-CSF in chronic stroke patients. This study thus provides the basis to explore the efficacy of G-CSF in improving chronic stroke related deficits. TRIAL REGISTRATION: ClinicalTrials.gov NCT00298597. PMID- 21625427 TI - Loss of the SPHF homologue Slr1768 leads to a catastrophic failure in the maintenance of thylakoid membranes in Synechocystis sp. PCC 6803. AB - BACKGROUND: In cyanobacteria the photosystems are localised to, and maintained in, specialist membranes called the thylakoids. The mechanism driving the biogenesis of the thylakoid membranes is still an open question, with only two potential biogenesis factors, Vipp1 and Alb3 currently identified. METHODOLOGY/PRINCIPAL FINDINGS: We generated a slr1768 knockout using the pGEM T easy vector and REDIRECT. By comparing growth and pigment content (chlorophyll a fluoresence) of the Deltaslr1768 mutant with the wild-type, we found that Deltaslr1768 has a conditional phenotype; specifically under high light conditions (130 umol m(-2) s(-1)) thylakoid biogenesis is disrupted leading to cell death on a scale of days. The thylakoids show considerable disruption, with loss of both structure and density, while chlorophyll a density decreases with the loss of thylakoids, although photosynthetic efficiency is unaffected. Under low light (30 umol m(-2) s(-1)) the phenotype is significantly reduced, with a growth rate similar to the wild-type and only a low frequency of cells with evident thylakoid disruption. CONCLUSIONS/SIGNIFICANCE: This is the first example of a gene that affects the maintenance of the thylakoid membranes specifically under high light, and which displays a phenotype dependent on light intensity. Our results demonstrate that Slr1768 has a leading role in acclimatisation, linking light damage with maintenance of the thylakoids. PMID- 21625428 TI - Do postures of distal effectors affect the control of actions of other distal effectors? Evidence for a system of interactions between hand and mouth. AB - The present study aimed at determining whether, in healthy humans, postures assumed by distal effectors affect the control of the successive grasp executed with other distal effectors. In experiments 1 and 2, participants reached different objects with their head and grasped them with their mouth, after assuming different hand postures. The postures could be implicitly associated with interactions with large or small objects. The kinematics of lip shaping during grasp varied congruently with the hand posture, i.e. it was larger or smaller when it could be associated with the grasping of large or small objects, respectively. In experiments 3 and 4, participants reached and grasped different objects with their hand, after assuming the postures of mouth aperture or closure (experiment 3) and the postures of toe extension or flexion (experiment 4). The mouth postures affected the kinematics of finger shaping during grasp, that is larger finger shaping corresponded with opened mouth and smaller finger shaping with closed mouth. In contrast, the foot postures did not influence the hand grasp kinematics. Finally, in experiment 5 participants reached-grasped different objects with their hand while pronouncing opened and closed vowels, as verified by the analysis of their vocal spectra. Open and closed vowels induced larger and smaller finger shaping, respectively. In all experiments postures of the distal effectors induced no effect, or only unspecific effects on the kinematics of the reach proximal/axial component. The data from the present study support the hypothesis that there exists a system involved in establishing interactions between movements and postures of hand and mouth. This system might have been used to transfer a repertoire of hand gestures to mouth articulation postures during language evolution and, in modern humans, it may have evolved a system controlling the interactions existing between speech and gestures. PMID- 21625429 TI - Short-snouted toothless ichthyosaur from China suggests Late Triassic diversification of suction feeding ichthyosaurs. AB - BACKGROUND: Ichthyosaurs were an important group of Mesozoic marine reptiles and existed from the Early Triassic to the early Late Cretaceous. Despite a great diversity in body shapes and feeding adaptations, all share greatly enlarged eyes, an elongated rostrum with numerous conical teeth, and a streamlined body. METHODOLOGY/PRINCIPAL FINDINGS: Based on new material from China and the restudy of Shastasaurus pacificus, we here reinterpret the classical large-bodied Late Triassic ichthyosaur genus Shastasaurus to differ greatly from the standard ichthyosaurian body plan, indicating much greater morphological diversity and range of feeding adaptations in ichthyosaurs than previously recognized. Phylogenetic analysis indicates a monophyletic clade consisting of the giant Shonisaurus sikanniensis, Guanlingsaurus liangae, and Shastasaurus pacificus to which the genus name Shastasaurus is applied. Shastasaurus liangae comb. nov. is from the Late Triassic (Carnian) Xiaowa Formation of Guizhou Province, southwestern China. The species combines a diminutive head with an entirely toothless and greatly reduced snout. The species also has by far the highest vertebral count among ichthyosaurs (86 presacral vertebrae and >110 caudal vertebrae), a count that is also very high for tetrapods in general. A reduced toothless snout and a diminutive head is also apparently present in the giant S. sikanniensis and presumably in S. pacificus. CONCLUSIONS/SIGNIFICANCE: In analogy to many modern odontocetes, Shastasaurus is interpreted as a specialized suction feeder on unshelled cephalopods and fish, suggesting a unique but widespread Late Triassic diversification of toothless, suction-feeding ichthyosaurs. Suction feeding has not been hypothesized for any of the other diverse marine reptiles of the Mesozoic before, but in Shastasaurus may be linked to the Late Triassic minimum in atmospheric oxygen. PMID- 21625430 TI - Reorganization of functional networks in mild cognitive impairment. AB - Whether the balance between integration and segregation of information in the brain is damaged in Mild Cognitive Impairment (MCI) subjects is still a matter of debate. Here we characterize the functional network architecture of MCI subjects by means of complex networks analysis. Magnetoencephalograms (MEG) time series obtained during a memory task were evaluated by synchronization likelihood (SL), to quantify the statistical dependence between MEG signals and to obtain the functional networks. Graphs from MCI subjects show an enhancement of the strength of connections, together with an increase in the outreach parameter, suggesting that memory processing in MCI subjects is associated with higher energy expenditure and a tendency toward random structure, which breaks the balance between integration and segregation. All features are reproduced by an evolutionary network model that simulates the degenerative process of a healthy functional network to that associated with MCI. Due to the high rate of conversion from MCI to Alzheimer Disease (AD), these results show that the analysis of functional networks could be an appropriate tool for the early detection of both MCI and AD. PMID- 21625431 TI - Current and future patterns of global marine mammal biodiversity. AB - Quantifying the spatial distribution of taxa is an important prerequisite for the preservation of biodiversity, and can provide a baseline against which to measure the impacts of climate change. Here we analyse patterns of marine mammal species richness based on predictions of global distributional ranges for 115 species, including all extant pinnipeds and cetaceans. We used an environmental suitability model specifically designed to address the paucity of distributional data for many marine mammal species. We generated richness patterns by overlaying predicted distributions for all species; these were then validated against sightings data from dedicated long-term surveys in the Eastern Tropical Pacific, the Northeast Atlantic and the Southern Ocean. Model outputs correlated well with empirically observed patterns of biodiversity in all three survey regions. Marine mammal richness was predicted to be highest in temperate waters of both hemispheres with distinct hotspots around New Zealand, Japan, Baja California, the Galapagos Islands, the Southeast Pacific, and the Southern Ocean. We then applied our model to explore potential changes in biodiversity under future perturbations of environmental conditions. Forward projections of biodiversity using an intermediate Intergovernmental Panel for Climate Change (IPCC) temperature scenario predicted that projected ocean warming and changes in sea ice cover until 2050 may have moderate effects on the spatial patterns of marine mammal richness. Increases in cetacean richness were predicted above 40 degrees latitude in both hemispheres, while decreases in both pinniped and cetacean richness were expected at lower latitudes. Our results show how species distribution models can be applied to explore broad patterns of marine biodiversity worldwide for taxa for which limited distributional data are available. PMID- 21625432 TI - The interplay between NF-kappaB and E2F1 coordinately regulates inflammation and metabolism in human cardiac cells. AB - Pyruvate dehydrogenase kinase 4 (PDK4) inhibition by nuclear factor-kappaB (NF kappaB) is related to a shift towards increased glycolysis during cardiac pathological processes such as cardiac hypertrophy and heart failure. The transcription factors estrogen-related receptor-alpha (ERRalpha) and peroxisome proliferator-activated receptor (PPAR) regulate PDK4 expression through the potent transcriptional coactivator PPARgamma coactivator-1alpha (PGC-1alpha). NF kappaB activation in AC16 cardiac cells inhibit ERRalpha and PPARbeta/delta transcriptional activity, resulting in reduced PGC-1alpha and PDK4 expression, and an enhanced glucose oxidation rate. However, addition of the NF-kappaB inhibitor parthenolide to these cells prevents the downregulation of PDK4 expression but not ERRalpha and PPARbeta/delta DNA binding activity, thus suggesting that additional transcription factors are regulating PDK4. Interestingly, a recent study has demonstrated that the transcription factor E2F1, which is crucial for cell cycle control, may regulate PDK4 expression. Given that NF-kappaB may antagonize the transcriptional activity of E2F1 in cardiac myocytes, we sought to study whether inflammatory processes driven by NF kappaB can downregulate PDK4 expression in human cardiac AC16 cells through E2F1 inhibition. Protein coimmunoprecipitation indicated that PDK4 downregulation entailed enhanced physical interaction between the p65 subunit of NF-kappaB and E2F1. Chromatin immunoprecipitation analyses demonstrated that p65 translocation into the nucleus prevented the recruitment of E2F1 to the PDK4 promoter and its subsequent E2F1-dependent gene transcription. Interestingly, the NF-kappaB inhibitor parthenolide prevented the inhibition of E2F1, while E2F1 overexpression reduced interleukin expression in stimulated cardiac cells. Based on these findings, we propose that NF-kappaB acts as a molecular switch that regulates E2F1-dependent PDK4 gene transcription. PMID- 21625433 TI - Transplantation of neuronal-primed human bone marrow mesenchymal stem cells in hemiparkinsonian rodents. AB - Bone marrow-derived human mesenchymal stem cells (hMSCs) have shown promise in in vitro neuronal differentiation and in cellular therapy for neurodegenerative disorders, including Parkinson' disease. However, the effects of intracerebral transplantation are not well defined, and studies do not agreed on the optimal neuronal differentiation method. Here, we investigated three growth factor-based neuronal differentiation procedures (using FGF-2/EGF/PDGF/SHH/FGF-8/GDNF), and found all to be capable of eliciting an immature neural phenotype, in terms of cell morphology and gene/protein expression. The neuronal-priming (FGF-2/EGF) method induced neurosphere-like formation and the highest NES and NR4A2 expression by hMSCs. Transplantation of undifferentiated and neuronal-primed hMSCs into the striatum and substantia nigra of 6-OHDA-lesioned hemiparkinsonian rats revealed transient graft survival of 7 days, despite the reported immunosuppressive properties of MSCs and cyclosporine-immunosuppression of rats. Neither differentiation of hMSCs nor induction of host neurogenesis was observed at injection sites, and hMSCs continued producing mesodermal fibronectin. Strategies for improving engraftment and differentiation post-transplantation, such as prior in vitro neuronal-priming, nigral and striatal grafting, and co transplantation of olfactory ensheathing cells that promote neural regeneration, were unable to provide advantages. Innate inflammatory responses (Iba-1-positive microglia/macrophage and GFAP-positive astrocyte activation and accumulation) were detected around grafts within 7 days. Our findings indicate that growth factor-based methods allow hMSC differentiation toward immature neuronal-like cells, and contrary to previous reports, only transient survival and engraftment of hMSCs occurs following transplantation in immunosuppressed hemiparkinsonian rats. In addition, suppression of host innate inflammatory responses may be a key factor for improving hMSC survival and engraftment. PMID- 21625434 TI - Non-integrative lentivirus drives high-frequency cre-mediated cassette exchange in human cells. AB - Recombinase mediated cassette exchange (RMCE) is a two-step process leading to genetic modification in a specific genomic target sequence. The process involves insertion of a docking genetic cassette in the genome followed by DNA transfer of a second cassette flanked by compatible recombination signals and expression of the recombinase. Major technical drawbacks are cell viability upon transfection, toxicity of the enzyme, and the ability to target efficiently cell types of different origins. To overcome such drawbacks, we developed an RMCE assay that uses an integrase-deficient lentivirus (IDLV) vector in the second step combined with promoterless trapping of double selectable markers. Additionally, recombinase expression is self-limiting as a result of the exchangeable reaction, thus avoiding toxicity. Our approach provides proof-of-principle of a simple and novel strategy with expected wide applicability modelled on a human cell line with randomly integrated copies of a genetic landing pad. This strategy does not present foreseeable limitations for application to other cell systems modified by homologous recombination. Safety, efficiency, and simplicity are the major advantages of our system, which can be applied in low-to-medium throughput strategies for screening of cDNAs, non-coding RNAs during functional genomic studies, and drug screening. PMID- 21625435 TI - Growth differentiation factor 15 is induced by hepatitis C virus infection and regulates hepatocellular carcinoma-related genes. AB - Liver fibrosis, cirrhosis, and hepatocellular carcinoma (HCC) are commonly induced by chronic hepatitis C virus (HCV) infection. We aimed to identify and characterize the involvement of previously screened cytokine GDF15 in HCV pathogenesis. We examined the GDF15 expression after HCV infection both in vitro and in vivo. Cultured JFH-1 HCV was used to determine the GDF15 function on virus propagation. GDF15 overexpression and RNA interference were employed to profile the GDF15-regulated genes, signaling pathways and cell biology phenotypes. The mRNA expression and protein secretion of GDF15 was dramatically increased in HCV infected hepatoma cells, which maybe a host response to viral proteins or infection-induced cell stress. Patients infected with HCV had an average 15-fold higher blood GDF15 level than that of healthy volunteers. Three HCC individuals in the HCV cohort showed extremely high GDF15 concentrations. Transfection or exogenously supplied GDF15 enhanced HCV propagation, whereas knockdown of endogenous GDF15 resulted in inhibition of virus replication. Overexpressed GDF15 led to Akt activation and the phosphorylation of Akt downstream targeted GSK 3beta and Raf. Several HCC-related molecules, such as E-cadherin, beta-catenin, Cyclin A2/B1/D1, were up-regulated by GDF15 stimulation in vitro. Overexpression of GDF15 in hepatoma cells resulted in increased DNA synthesis, promoted cell proliferation, and importantly enhanced invasiveness of the cells. In conclusion, these results suggest that an elevated serum GDF15 level is a potential diagnostic marker for viral hepatitis, and GDF15 may contribute to HCV pathogenesis by altering the signaling and growth of host cells. PMID- 21625436 TI - NOA1 functions in a temperature-dependent manner to regulate chlorophyll biosynthesis and Rubisco formation in rice. AB - NITRIC OXIDE-ASSOCIATED1 (NOA1) encodes a circularly permuted GTPase (cGTPase) known to be essential for ribosome assembly in plants. While the reduced chlorophyll and Rubisco phenotypes were formerly noticed in both NOA1-suppressed rice and Arabidopsis, a detailed insight is still necessary. In this study, by using RNAi transgenic rice, we further demonstrate that NOA1 functions in a temperature-dependent manner to regulate chlorophyll and Rubisco levels. When plants were grown at 30 degrees C, the chlorophyll and Rubisco levels in OsNOA1 silenced plants were only slightly lower than those in WT. However, at 22 degrees C, the silenced plants accumulated far less chlorophyll and Rubisco than WT. It was further revealed that the regulation of chlorophyll and Rubisco occurs at the anabolic level. Etiolated WT seedlings restored chlorophyll and Rubisco accumulations readily once returned to light, at either 30 degrees C or 15 degrees C. Etiolated OsNOA1-silenced plants accumulated chlorophyll and Rubisco to normal levels only at 30 degrees C, and lost this ability at low temperature. On the other hand, de-etiolated OsNOA1-silenced seedlings maintained similar levels of chlorophyll and Rubisco as WT, even after being shifted to 15 degrees C for various times. Further expression analyses identified several candidate genes, including OsPorA (NADPH: protochlorophyllide oxidoreductase A), OsrbcL (Rubisco large subunit), OsRALyase (Ribosomal RNA apurinic site specific lyase) and OsPuf4 (RNA-binding protein of the Puf family), which may be involved in OsNOA1-regulated chlorophyll biosynthesis and Rubisco formation. Overall, our results suggest OsNOA1 functions in a temperature-dependent manner to regulate chlorophyll biosynthesis, Rubisco formation and plastid development in rice. PMID- 21625437 TI - Intermittent hypoxia-induced cognitive deficits are mediated by NADPH oxidase activity in a murine model of sleep apnea. AB - BACKGROUND: In rodents, exposure to intermittent hypoxia (IH), a hallmark of obstructive sleep apnea (OSA), is associated with neurobehavioral impairments, increased apoptosis in the hippocampus and cortex, as well as increased oxidant stress and inflammation. Excessive NADPH oxidase activity may play a role in IH induced CNS dysfunction. METHODS AND FINDINGS: The effect of IH during light period on two forms of spatial learning in the water maze and well as markers of oxidative stress was assessed in mice lacking NADPH oxidase activity (gp91phox(_/Y)) and wild-type littermates. On a standard place training task, gp91phox(_/Y) displayed normal learning, and were protected from the spatial learning deficits observed in wild-type littermates exposed to IH. Moreover, anxiety levels were increased in wild-type mice exposed to IH as compared to room air (RA) controls, while no changes emerged in gp91phox(_/Y) mice. Additionally, wild-type mice, but not gp91phox(_/Y) mice had significantly elevated levels of NADPH oxidase expression and activity, as well as MDA and 8-OHDG in cortical and hippocampal lysates following IH exposures. CONCLUSIONS: The oxidative stress responses and neurobehavioral impairments induced by IH during sleep are mediated, at least in part, by excessive NADPH oxidase activity, and thus pharmacological agents targeting NADPH oxidase may provide a therapeutic strategy in sleep-disordered breathing. PMID- 21625438 TI - PIF genes mediate the effect of sucrose on seedling growth dynamics. AB - As photoautotrophs, plants can use both the form and amount of fixed carbon as a measure of the light environment. In this study, we used a variety of approaches to elucidate the role of exogenous sucrose in modifying seedling growth dynamics. In addition to its known effects on germination, high-resolution temporal analysis revealed that sucrose could extend the number of days plants exhibited rapid hypocotyl elongation, leading to dramatic increases in ultimate seedling height. In addition, sucrose changed the timing of daily growth maxima, demonstrating that diel growth dynamics are more plastic than previously suspected. Sucrose-dependent growth promotion required function of multiple phytochrome-interacting factors (PIFs), and overexpression of PIF5 led to growth dynamics similar to plants exposed to sucrose. Consistent with this result, sucrose was found to increase levels of PIF5 protein. PIFs have well-established roles as integrators of response to light levels, time of day and phytohormone signaling. Our findings strongly suggest that carbon availability can modify the known photomorphogenetic signaling network. PMID- 21625439 TI - Cleavage of kininogen and subsequent bradykinin release by the complement component: mannose-binding lectin-associated serine protease (MASP)-1. AB - Bradykinin (BK), generated from high-molecular-weight kininogen (HK) is the major mediator of swelling attacks in hereditary angioedema (HAE), a disease associated with C1-inhibitor deficiency. Plasma kallikrein, activated by factor XIIa, is responsible for most of HK cleavage. However other proteases, which activate during episodes of angioedema, might also contribute to BK production. The lectin pathway of the complement system activates after infection and oxidative stress on endothelial cells generating active serine proteases: MASP-1 and MASP-2. Our aim was to study whether activated MASPs are able to digest HK to release BK. Initially we were trying to find potential new substrates of MASP-1 in human plasma by differential gel electrophoresis, and we identified kininogen cleavage products by this proteomic approach. As a control, MASP-2 was included in the study in addition to MASP-1 and kallikrein. The proteolytic cleavage of HK by MASPs was followed by SDS-PAGE, and BK release was detected by HPLC. We showed that MASP-1 was able to cleave HK resulting in BK production. MASP-2 could also cleave HK but could not release BK. The cleavage pattern of MASPs is similar but not strictly identical to that of kallikrein. The catalytic efficiency of HK cleavage by a recombinant version of MASP-1 and MASP-2 was about 4.0*10(2) and 2.7*10(2) M(-1) s(-1), respectively. C1-inhibitor, the major inhibitor of factor XIIa and kallikrein, also prevented the cleavage of HK by MASPs. In all, a new factor XII- and kallikrein-independent mechanism of bradykinin production by MASP 1 was demonstrated, which may contribute to the pro-inflammatory effect of the lectin pathway of complement and to the elevated bradykinin levels in HAE patients. PMID- 21625440 TI - Study of histopathological and molecular changes of rat kidney under simulated weightlessness and resistance training protective effect. AB - To explore the effects of long-term weightlessness on the renal tissue, we used the two months tail suspension model to simulate microgravity and investigated the simulated microgravity on the renal morphological damages and related molecular mechanisms. The microscopic examination of tissue structure and ultrastructure was carried out for histopathological changes of renal tissue morphology. The immunohistochemistry, real-time PCR and Western blot were performed to explore the molecular mechanisms associated the observations. Hematoxylin and eosin (HE) staining showed severe pathological kidney lesions including glomerular atrophy, degeneration and necrosis of renal tubular epithelial cells in two months tail-suspended rats. Ultrastructural studies of the renal tubular epithelial cells demonstrated that basal laminas of renal tubules were rough and incrassate with mitochondria swelling and vacuolation. Cell apoptosis in kidney monitored by the expression of Bax/Bcl-2 and caspase-3 accompanied these pathological damages caused by long-term microgravity. Analysis of the HSP70 protein expression illustrated that overexpression of HSP70 might play a crucial role in inducing those pathological damages. Glucose regulated protein 78 (GRP78), one of the endoplasmic reticulum (ER) chaperones, was up regulated significantly in the kidney of tail suspension rat, which implied that ER-stress was associated with apoptosis. Furthermore, CHOP and caspase-12 pathways were activated in ER-stress induced apoptosis. Resistance training not only reduced kidney cell apoptosis and expression of HSP70 protein, it also can attenuate the kidney impairment imposed by weightlessness. The appropriate optimization might be needed for the long term application for space exploration. PMID- 21625441 TI - Screen for IDH1, IDH2, IDH3, D2HGDH and L2HGDH mutations in glioblastoma. AB - Isocitrate dehydrogenases (IDHs) catalyse oxidative decarboxylation of isocitrate to alpha-ketoglutarate (alpha-KG). IDH1 functions in the cytosol and peroxisomes, whereas IDH2 and IDH3 are both localized in the mitochondria. Heterozygous somatic mutations in IDH1 occur at codon 132 in 70% of grade II-III gliomas and secondary glioblastomas (GBMs), and in 5% of primary GBMs. Mutations in IDH2 at codon 172 are present in grade II-III gliomas at a low frequency. IDH1 and IDH2 mutations cause both loss of normal enzyme function and gain-of-function, causing reduction of alpha-KG to D-2-hydroxyglutarate (D-2HG) which accumulates. Excess hydroxyglutarate (2HG) can also be caused by germline mutations in D- and L-2 hydroxyglutarate dehydrogenases (D2HGDH and L2HGDH). If loss of IDH function is critical for tumourigenesis, we might expect some tumours to acquire somatic IDH3 mutations. Alternatively, if 2HG accumulation is critical, some tumours might acquire somatic D2HGDH or L2HGDH mutations. We therefore screened 47 glioblastoma samples looking for changes in these genes. Although IDH1 R132H was identified in 12% of samples, no mutations were identified in any of the other genes. This suggests that mutations in IDH3, D2HGDH and L2HGDH do not occur at an appreciable frequency in GBM. One explanation is simply that mono-allelic IDH1 and IDH2 mutations occur more frequently by chance than the bi-allelic mutations expected at IDH3, D2HGDH and L2HGDH. Alternatively, both loss of IDH function and 2HG accumulation might be required for tumourigenesis, and only IDH1 and IDH2 mutations have these dual effects. PMID- 21625442 TI - Methylation profile of single hepatocytes derived from hepatitis B virus-related hepatocellular carcinoma. AB - BACKGROUND: With the development of high-throughput screening, a variety of genetic alterations has been found in hepatocellular carcinoma (HCC). Although previous studies on HCC methylation profiles have focused on liver tissue, studies using isolated hepatocytes are rare. The heterogeneity of liver composition may impact the genuine methylation status of HCC; therefore, it is important to clarify the methylation profile of hepatocytes to aid in understanding the process of tumorigenesis. METHODS AND FINDINGS: The global methylation profile of single hepatocytes isolated from liver tissue of hepatitis B virus (HBV) related HCC (HBHC) was analyzed using Illumina Infinium Human Methylation27 BeadChips, and combined bisulfite restriction analysis (COBRA) and bisulfite sequencing were used to validate the 20 significant hypermethylated genes identified. In this study, we found many noteworthy differences in the genome-wide methylation profiles of single hepatocytes of HBHC. Unsupervised hierarchical clustering analysis showed that hepatocyte methylation profiles could be classified according to three cell types: hepatocytes of HCC, adjacent hepatocytes and normal hepatocytes. Among the 20 most hypermethylated genes in the hepatocytes of HBHC, 7 novel genes (WNK2, EMILIN2, TLX3, TM6SF1, TRIM58, HIST1H4Fand GRASP) were found to be hypermethylated in HBHC and hypomethylated in paired adjacent liver tissues; these findings have not been reported in previous studies on tissue samples. CONCLUSION: The genome-wide methylation profile of purified single hepatocytes of HBHC was aided in understanding the process of tumorigenesis, and a series of novel methylated genes found in this study have the potential to be biomarkers for the diagnosis and prognosis of HBHC. PMID- 21625443 TI - A locus on chromosome 5 is associated with dilated cardiomyopathy in Doberman Pinschers. AB - Dilated cardiomyopathy (DCM) is a heterogeneous group of heart diseases with a strong genetic background. Currently, many human DCM cases exist where no causative mutation can be identified. DCM also occurs with high prevalence in several large dog breeds. In the Doberman Pinscher a specific DCM form characterized by arrhythmias and/or echocardiographic changes has been intensively studied by veterinary cardiologists. We performed a genome-wide association study in Doberman Pinschers. Using 71 cases and 70 controls collected in Germany we identified a genome-wide significant association to DCM on chromosome 5. We validated the association in an independent cohort collected in the United Kingdom. There is no known DCM candidate gene under the association signal. Therefore, DCM in Doberman Pinschers offers the chance of identifying a novel DCM gene that might also be relevant for human health. PMID- 21625444 TI - Short-term synaptic plasticity in the dentate gyrus of monkeys. AB - The hippocampus plays an important role in learning and memory. Synaptic plasticity in the hippocampus, short-term and long-term, is postulated to be a neural substrate of memory trace. Paired-pulse stimulation is a standard technique for evaluating a form of short-term synaptic plasticity in rodents. However, evidence is lacking for paired-pulse responses in the primate hippocampus. In the present study, we recorded paired-pulse responses in the dentate gyrus of monkeys while stimulating to the medial part of the perforant path at several inter-pulse intervals (IPIs) using low and high stimulus intensities. When the stimulus intensity was low, the first pulse produced early strong depression (at IPIs of 10-30 ms) and late slight depression (at IPIs of 100-1000 ms) of field excitatory postsynaptic potentials (fEPSPs) generated by the second pulse, interposing no depression IPIs (50-70 ms). When the stimulus intensity was high, fEPSPs generated by the second pulse were depressed by the first pulse at all IPIs except for the longest one (2000 ms). Population spikes (PSs) generated by the second pulse were completely blocked or strongly depressed at shorter IPIs (10-100 or 200 ms, respectively), while no depression or slight facilitation occurred at longer IPIs (500-2000 ms). Administration of diazepam slightly increased fEPSPs, while it decreased PSs produced by the first pulse. It also enhanced the facilitation of PSs produced by the second stimulation at longer IPIs. The present results, in comparison with previous studies using rodents, indicate that paired-pulse responses of fEPSPs in the monkey are basically similar to those of rodents, although paired-pulse responses of PSs in the monkey are more delayed than those in rodents and have a different sensitivity to diazepam. PMID- 21625445 TI - Cortical processing of swallowing in ALS patients with progressive dysphagia--a magnetoencephalographic study. AB - Amyotrophic lateral sclerosis (ALS) is a rare disease causing degeneration of the upper and lower motor neuron. Involvement of the bulbar motor neurons often results in fast progressive dysphagia. While cortical compensation of dysphagia has been previously shown in stroke patients, this topic has not been addressed in patients suffering from ALS. In the present study, we investigated cortical activation during deglutition in two groups of ALS patients with either moderate or severe dysphagia. Whole-head MEG was employed on fourteen patients with sporadic ALS using a self-paced swallowing paradigm. Data were analyzed by means of time-frequency analysis and synthetic aperture magnetometry (SAM). Group analysis of individual SAM data was performed using a permutation test. We found a reduction of cortical swallowing related activation in ALS patients compared to healthy controls. Additionally a disease-related shift of hemispheric lateralization was observed. While healthy subjects showed bilateral cortical activation, the right sensorimotor cortex was predominantly involved in ALS patients. Both effects were even stronger in the group of patients with severe dysphagia. Our results suggest that bilateral degeneration of the upper motor neuron in the primary motor areas also impairs further adjusted motor areas, which leads to a strong reduction of 'swallowing related' cortical activation. While both hemispheres are affected by the degeneration a relatively stronger activation is seen in the right hemisphere. This right hemispheric lateralization of volitional swallowing observed in this study may be the only sign of cortical plasticity in dysphagic ALS patients. It may demonstrate compensational mechanisms in the right hemisphere which is known to predominantly coordinate the pharyngeal phase of deglutition. These results add new aspects to our understanding of the pathophysiology of dysphagia in ALS patients and beyond. The compensational mechanisms observed could be relevant for future research in swallowing therapies. PMID- 21625446 TI - Four small puzzles that Rosetta doesn't solve. AB - A complete macromolecule modeling package must be able to solve the simplest structure prediction problems. Despite recent successes in high resolution structure modeling and design, the Rosetta software suite fares poorly on small protein and RNA puzzles, some as small as four residues. To illustrate these problems, this manuscript presents Rosetta results for four well-defined test cases: the 20-residue mini-protein Trp cage, an even smaller disulfide-stabilized conotoxin, the reactive loop of a serine protease inhibitor, and a UUCG RNA tetraloop. In contrast to previous Rosetta studies, several lines of evidence indicate that conformational sampling is not the major bottleneck in modeling these small systems. Instead, approximations and omissions in the Rosetta all atom energy function currently preclude discriminating experimentally observed conformations from de novo models at atomic resolution. These molecular "puzzles" should serve as useful model systems for developers wishing to make foundational improvements to this powerful modeling suite. PMID- 21625447 TI - Secretome-based identification of ULBP2 as a novel serum marker for pancreatic cancer detection. AB - BACKGROUND: To discover novel markers for improving the efficacy of pancreatic cancer (PC) diagnosis, the secretome of two PC cell lines (BxPC-3 and MIA PaCa-2) was profiled. UL16 binding protein 2 (ULBP2), one of the proteins identified in the PC cell secretome, was selected for evaluation as a biomarker for PC detection because its mRNA level was also found to be significantly elevated in PC tissues. METHODS: ULBP2 expression in PC tissues from 67 patients was studied by immunohistochemistry. ULBP2 serum levels in 154 PC patients and 142 healthy controls were measured by bead-based immunoassay, and the efficacy of serum ULBP2 for PC detection was compared with the widely used serological PC marker carbohydrate antigen 19-9 (CA 19-9). RESULTS: Immunohistochemical analyses revealed an elevated expression of ULPB2 in PC tissues compared with adjacent non cancerous tissues. Meanwhile, the serum levels of ULBP2 among all PC patients (n = 154) and in early-stage cancer patients were significantly higher than those in healthy controls (p<0.0001). The combination of ULBP2 and CA 19-9 outperformed each marker alone in distinguishing PC patients from healthy individuals. Importantly, an analysis of the area under receiver operating characteristic curves showed that ULBP2 was superior to CA 19-9 in discriminating patients with early-stage PC from healthy controls. CONCLUSIONS: Collectively, our results indicate that ULBP2 may represent a novel and useful serum biomarker for pancreatic cancer primary screening. PMID- 21625448 TI - The metabolic consequences of hepatic AMP-kinase phosphorylation in rainbow trout. AB - AMP-activated protein kinase (AMPK), a phylogenetically conserved serine/threonine protein kinase, is proposed to function as a "fuel gauge" to monitor cellular energy status in response to nutritional environmental variations. However, in fish, few studies have addressed the metabolic consequences related to the activation of this kinase. This study demonstrates that the rainbow trout (Oncorhynchus mykiss) possesses paralogs of the three known AMPK subunits that co-diversified, that the AMPK protein is present in the liver and in isolated hepatocytes, and it does change in response to physiological (fasting-re-feeding cycle) and pharmacological (AICAR and metformin administration and incubations) manipulations. Moreover, the phosphorylation of AMPK results in the phosphorylation of acetyl-CoA carboxylase, a main downstream target of AMPK in mammals. Other findings include changes in hepatic glycogen levels and several molecular actors involved in hepatic glucose and lipid metabolism, including mRNA transcript levels for glucokinase, glucose-6 phosphatase and fatty acid synthase both in vivo and in vitro. The fact that most results presented in this study are consistent with the recognized role of AMPK as a master regulator of energy homeostasis in living organisms supports the idea that these functions are conserved in this piscine model. PMID- 21625449 TI - Theiler's murine encephalomyelitis virus as a vaccine candidate for immunotherapy. AB - The induction of sterilizing T-cell responses to tumors is a major goal in the development of T-cell vaccines for treating cancer. Although specific components of anti-viral CD8+ immunity are well characterized, we still lack the ability to mimic viral CD8+ T-cell responses in therapeutic settings for treating cancers. Infection with the picornavirus Theiler's murine encephalomyelitis virus (TMEV) induces a strong sterilizing CD8+ T-cell response. In the absence of sterilizing immunity, the virus causes a persistent infection. We capitalized on the ability of TMEV to induce strong cellular immunity even under conditions of immune deficiency by modifying the virus to evaluate its potential as a T-cell vaccine. The introduction of defined CD8+ T-cell epitopes into the leader sequence of the TMEV genome generates an attenuated vaccine strain that can efficiently drive CD8+ T-cell responses to the targeted antigen. This virus activates T-cells in a manner that is capable of inducing targeted tissue damage and glucose dysregulation in an adoptive T-cell transfer model of diabetes mellitus. As a therapeutic vaccine for the treatment of established melanoma, epitope-modified TMEV can induce strong cytotoxic T-cell responses and promote infiltration of the T-cells into established tumors, ultimately leading to a delay in tumor growth and improved survival of vaccinated animals. We propose that epitope-modified TMEV is an excellent candidate for further development as a human T-cell vaccine for use in immunotherapy. PMID- 21625450 TI - Time-ordered networks reveal limitations to information flow in ant colonies. AB - BACKGROUND: An important function of many complex networks is to inhibit or promote the transmission of disease, resources, or information between individuals. However, little is known about how the temporal dynamics of individual-level interactions affect these networks and constrain their function. Ant colonies are a model comparative system for understanding general principles linking individual-level interactions to network-level functions because interactions among individuals enable integration of multiple sources of information to collectively make decisions, and allocate tasks and resources. METHODOLOGY/FINDINGS: Here we show how the temporal and spatial dynamics of such individual interactions provide upper bounds to rates of colony-level information flow in the ant Temnothorax rugatulus. We develop a general framework for analyzing dynamic networks and a mathematical model that predicts how information flow scales with individual mobility and group size. CONCLUSIONS/SIGNIFICANCE: Using thousands of time-stamped interactions between uniquely marked ants in four colonies of a range of sizes, we demonstrate that observed maximum rates of information flow are always slower than predicted, and are constrained by regulation of individual mobility and contact rate. By accounting for the ordering and timing of interactions, we can resolve important difficulties with network sampling frequency and duration, enabling a broader understanding of interaction network functioning across systems and scales. PMID- 21625451 TI - Support of unrelated stem cell donor searches by donor center-initiated HLA typing of potentially matching donors. AB - Large registries of potential unrelated stem cell donors have been established in order to enable stem cell transplantation for patients without HLA-identical related donors. Donor search is complicated by the fact that the stored HLA information of many registered donors is incomplete. We carried out a project that was aimed to improve chances of patients with ongoing donor searches to find an HLA-matched unrelated donor. For that purpose, we carried out additional donor center-initiated HLA-DRB1 typing of donors who were only typed for the HLA loci A and B so far and were potential matches for patients in need of a stem cell transplant. In total, 8,861 donors were contacted for donor center-initiated HLA DRB1 typing within 1,089 donor searches. 12 of these donors have donated stem cells so far, 8 thereof for their respective target patients. We conclude that chances of patients with ongoing donor searches to find an HLA-matched unrelated donor can indeed be improved by donor-center initiated typing that is carried out in addition to the standard donor search process. Our results also raise questions regarding the appropriate use of incompletely typed donors within unrelated donor searches. PMID- 21625452 TI - Engineering of insulin receptor isoform-selective insulin analogues. AB - BACKGROUND: The insulin receptor (IR) exists in two isoforms, A and B, and the isoform expression pattern is tissue-specific. The C-terminus of the insulin B chain is important for receptor binding and has been shown to contact the IR just adjacent to the region where the A and B isoforms differ. The aim of this study was to investigate the importance of the C-terminus of the B chain in IR isoform binding in order to explore the possibility of engineering tissue-specific/liver specific insulin analogues. METHODOLOGY/PRINCIPAL FINDINGS: Insulin analogue libraries were constructed by total amino acid scanning mutagenesis. The relative binding affinities for the A and B isoform of the IR were determined by competition assays using scintillation proximity assay technology. Structural information was obtained by X-ray crystallography. Introduction of B25A or B25N mutations resulted in analogues with a 2-fold preference for the B compared to the A isoform, whereas the opposite was observed with a B25Y substitution. An acidic amino acid residue at position B27 caused an additional 2-fold selective increase in affinity for the receptor B isoform for analogues bearing a B25N mutation. Furthermore, the combination of B25H with either B27D or B27E also resulted in B isoform-preferential analogues (2-fold preference) even though the corresponding single mutation analogues displayed no differences in relative isoform binding affinity. CONCLUSIONS/SIGNIFICANCE: We have discovered a new class of IR isoform-selective insulin analogues with 2-4-fold differences in relative binding affinities for either the A or the B isoform of the IR compared to human insulin. Our results demonstrate that a mutation at position B25 alone or in combination with a mutation at position B27 in the insulin molecule confers IR isoform selectivity. Isoform-preferential analogues may provide new opportunities for developing insulin analogues with improved clinical benefits. PMID- 21625453 TI - TGF-beta suppresses beta-catenin-dependent tolerogenic activation program in dendritic cells. AB - The mechanisms that underlie the critical dendritic cell (DC) function in maintainance of peripheral immune tolerance are incompletely understood, although the beta-catenin signaling pathway is critical for this role. The molecular details by which beta-catenin signaling is regulated in DCs are unknown. Mechanical disruption of murine bone marrow-derived DC (BMDC) clusters activates DCs while maintaining their tolerogenic potential and this activation is associated with beta-catenin signaling, providing a useful model with which to explore tolerance-associated beta-catenin signaling in DCs. In this report, we demonstrate novel molecular features of the signaling events that control DC activation in response to mechanical stimulation. Non-canonical beta-catenin signaling is an essential component of this tolerogenic activation and is modulated by adhesion molecules, including integrins. This unique beta-catenin dependent signaling pathway is constitutively active at low levels, suggesting that mechanical stimulation is not necessarily required for induction of this unique activation program. We additionally find that the immunomodulatory cytokine TGF-beta antagonizes beta-catenin in DCs, thereby selectively suppressing signaling associated with tolerogenic DC activation while having no impact on LPS-induced, beta-catenin-independent immunogenic activation. These findings provide new molecular insight into the regulation of a critical signaling pathway for DC function in peripheral immune tolerance. PMID- 21625454 TI - Mechanism of estradiol-induced block of voltage-gated K+ currents in rat medial preoptic neurons. AB - The present study was conducted to characterize possible rapid effects of 17-beta estradiol on voltage-gated K(+) channels in preoptic neurons and, in particular, to identify the mechanisms by which 17-beta-estradiol affects the K(+) channels. Whole-cell currents from dissociated rat preoptic neurons were studied by perforated-patch recording. 17-beta-Estradiol rapidly (within seconds) and reversibly reduced the K(+) currents, showing an EC(50) value of 9.7 uM. The effect was slightly voltage dependent, but independent of external Ca(2+), and not sensitive to an estrogen-receptor blocker. Although 17-alpha-estradiol also significantly reduced the K(+) currents, membrane-impermeant forms of estradiol did not reduce the K(+) currents and other estrogens, testosterone and cholesterol were considerably less effective. The reduction induced by estradiol was overlapping with that of the K(V)-2-channel blocker r-stromatoxin-1. The time course of K(+) current in 17-beta-estradiol, with a time-dependent inhibition and a slight dependence on external K(+), suggested an open-channel block mechanism. The properties of block were predicted from a computational model where 17-beta estradiol binds to open K(+) channels. It was concluded that 17-beta-estradiol rapidly reduces voltage-gated K(+) currents in a way consistent with an open channel block mechanism. This suggests a new mechanism for steroid action on ion channels. PMID- 21625455 TI - High throughput determination of TGFbeta1/SMAD3 targets in A549 lung epithelial cells. AB - BACKGROUND: Transforming growth factor beta 1 (TGFbeta1) plays a major role in many lung diseases including lung cancer, pulmonary hypertension, and pulmonary fibrosis. TGFbeta1 activates a signal transduction cascade that results in the transcriptional regulation of genes in the nucleus, primarily through the DNA binding transcription factor SMAD3. The objective of this study is to identify genome-wide scale map of SMAD3 binding targets and the molecular pathways and networks affected by the TGFbeta1/SMAD3 signaling in lung epithelial cells. METHODOLOGY: We combined chromatin immunoprecipitation with human promoter region microarrays (ChIP-on-chip) along with gene expression microarrays to study global transcriptional regulation of the TGFbeta1/SMAD3 pathway in human A549 alveolar epithelial cells. The molecular pathways and networks associated with TGFbeta1/SMAD3 signaling were identified using computational approaches. Validation of selected target gene expression and direct binding of SMAD3 to promoters were performed by quantitative real time RT-PCR and electrophoretic mobility shift assay on A549 and human primary lung epithelial cells. RESULTS AND CONCLUSIONS: Known TGFbeta1 target genes such as SERPINE1, SMAD6, SMAD7, TGFB1 and LTBP3, were found in both ChIP-on-chip and gene expression analyses as well as some previously unrecognized targets such as FOXA2. SMAD3 binding of FOXA2 promoter and changed expression were confirmed. Computational approaches combining ChIP-on-chip and gene expression microarray revealed multiple target molecular pathways affected by the TGFbeta1/SMAD3 signaling. Identification of global targets and molecular pathways and networks associated with TGFbeta1/SMAD3 signaling allow for a better understanding of the mechanisms that determine epithelial cell phenotypes in fibrogenesis and carcinogenesis as does the discovery of the direct effect of TGFbeta1 on FOXA2. PMID- 21625456 TI - Continuum molecular simulation of large conformational changes during ion-channel gating. AB - A modeling framework was developed to simulate large and gradual conformational changes within a macromolecule (protein) when its low amplitude high frequency vibrations are not concerned. Governing equations were derived as alternative to Langevin and Smoluchowski equations and used to simulate gating conformational changes of the Kv7.1 ion-channel over the time scale of its gating process (tens of milliseconds). The alternative equations predict the statistical properties of the motion trajectories with good accuracy and do not require the force field to be constant over the diffusion length, as assumed in Langevin equation. The open probability of the ion-channel was determined considering cooperativity of four subunits and solving their concerted transition to the open state analytically. The simulated open probabilities for a series of voltage clamp tests produced current traces that were similar to experimentally recorded currents. PMID- 21625457 TI - Variation in NOD2 augments Th2- and Th17 responses to myelin basic protein in multiple sclerosis. AB - Variations in the gene for the nucleotide-binding oligomerisation domain (NOD) 2 have been associated with Crohn's disease but not multiple sclerosis (MS). Here we investigate the effect of three polymorphisms in the NOD2 gene (rs5743277, rs2066842 and rs5743291) on cytokine production and CD4+ T cell proliferation elicited by human myelin basic protein (MBP) in blood mononuclear cell (MNC) cultures from 29 patients with MS. No polymorphism was observed at rs5743277. No associations with the rs2066842 polymorphism were found. Concerning rs5743291, none were homozygous for the minor allele. Seven of 29 (24%) patients were heterozygous, and five of these (71%) exhibited increased MBP-induced CD4+ T cell proliferation versus four of 22 (18%), who were homozygous for the major allele (p<0.04). Interleukin (IL)-5 was induced by MBP in MNC from the same five carriers versus two (9%) homozygotes (p<0.004); four carriers (57%) versus three non-carriers (14%) exhibited IL-17 responses to MBP (p<0.04). By contrast, we found no association between the polymorphisms investigated and interferon-gamma , tumor necrosis factor-alpha-, IL-2, -4- or IL-10 responses to MBP. These results indicate that the rs5743291 polymorphism influences T helper (Th) cell 2- and Th17 cell responses in MNC from MS patients. PMID- 21625458 TI - Thermostable direct hemolysin downregulates human colon carcinoma cell proliferation with the involvement of E-cadherin, and beta-catenin/Tcf-4 signaling. AB - BACKGROUND: Colon cancers are the frequent causes of cancer mortality worldwide. Recently bacterial toxins have received marked attention as promising approaches in the treatment of colon cancer. Thermostable direct hemolysin (TDH) secreted by Vibrio parahaemolyticus causes influx of extracellular calcium with the subsequent rise in intracellular calcium level in intestinal epithelial cells and it is known that calcium has antiproliferative activity against colon cancer. KEY RESULTS: In the present study it has been shown that TDH, a well-known traditional virulent factor inhibits proliferation of human colon carcinoma cells through the involvement of CaSR in its mechanism. TDH treatment does not induce DNA fragmentation, nor causes the release of lactate dehydrogenase. Therefore, apoptosis and cytotoxicity are not contributing to the TDH-mediated reduction of proliferation rate, and hence the reduction appears to be caused by decrease in cell proliferation. The elevation of E-cadherin, a cell adhesion molecule and suppression of beta-catenin, a proto-oncogene have been observed in presence of CaSR agonists whereas reverse effect has been seen in presence of CaSR antagonist as well as si-RNA in TDH treated cells. TDH also triggers a significant reduction of Cyclin-D and cdk2, two important cell cycle regulatory proteins along with an up regulation of cell cycle inhibitory protein p27(Kip1) in presence of CaSR agonists. CONCLUSION: Therefore TDH can downregulate colonic carcinoma cell proliferation and involves CaSR in its mechanism of action. The downregulation occurs mainly through the involvement of E-cadherin-beta-catenin mediated pathway and the inhibition of cell cycle regulators as well as upregulation of cell cycle inhibitors. PMID- 21625459 TI - Effect of anthropogenic landscape features on population genetic differentiation of Przewalski's gazelle: main role of human settlement. AB - Anthropogenic landscapes influence evolutionary processes such as population genetic differentiation, however, not every type of landscape features exert the same effect on a species, hence it is necessary to estimate their relative effect for species management and conservation. Przewalski's gazelle (Procapra przewalskii), which inhabits a human-altered area on Qinghai-Tibet Plateau, is one of the most endangered antelope species in the world. Here, we report a landscape genetic study on Przewalski's gazelle. We used skin and fecal samples of 169 wild gazelles collected from nine populations and thirteen microsatellite markers to assess the genetic effect of anthropogenic landscape features on this species. For comparison, the genetic effect of geographical distance and topography were also evaluated. We found significant genetic differentiation, six genetic groups and restricted dispersal pattern in Przewalski's gazelle. Topography, human settlement and road appear to be responsible for observed genetic differentiation as they were significantly correlated with both genetic distance measures [F(ST)/(1-F(ST)) and F'(ST)/(1-F'(ST))] in Mantel tests. IBD (isolation by distance) was also inferred as a significant factor in Mantel tests when genetic distance was measured as F(ST)/(1-F(ST)). However, using partial Mantel tests, AIC(c) calculations, causal modeling and AMOVA analysis, we found that human settlement was the main factor shaping current genetic differentiation among those tested. Altogether, our results reveal the relative influence of geographical distance, topography and three anthropogenic landscape-type on population genetic differentiation of Przewalski's gazelle and provide useful information for conservation measures on this endangered species. PMID- 21625460 TI - Sequence diversities of serine-aspartate repeat genes among Staphylococcus aureus isolates from different hosts presumably by horizontal gene transfer. AB - BACKGROUND: Horizontal gene transfer (HGT) is recognized as one of the major forces for bacterial genome evolution. Many clinically important bacteria may acquire virulence factors and antibiotic resistance through HGT. The comparative genomic analysis has become an important tool for identifying HGT in emerging pathogens. In this study, the Serine-Aspartate Repeat (Sdr) family has been compared among different sources of Staphylococcus aureus (S. aureus) to discover sequence diversities within their genomes. METHODOLOGY/PRINCIPAL FINDINGS: Four sdr genes were analyzed for 21 different S. aureus strains and 218 mastitis associated S. aureus isolates from Canada. Comparative genomic analyses revealed that S. aureus strains from bovine mastitis (RF122 and mastitis isolates in this study), ovine mastitis (ED133), pig (ST398), chicken (ED98), and human methicillin-resistant S. aureus (MRSA) (TCH130, MRSA252, Mu3, Mu50, N315, 04 02981, JH1 and JH9) were highly associated with one another, presumably due to HGT. In addition, several types of insertion and deletion were found in sdr genes of many isolates. A new insertion sequence was found in mastitis isolates, which was presumably responsible for the HGT of sdrC gene among different strains. Moreover, the sdr genes could be used to type S. aureus. Regional difference of sdr genes distribution was also indicated among the tested S. aureus isolates. Finally, certain associations were found between sdr genes and subclinical or clinical mastitis isolates. CONCLUSIONS: Certain sdr gene sequences were shared in S. aureus strains and isolates from different species presumably due to HGT. Our results also suggest that the distributional assay of virulence factors should detect the full sequences or full functional regions of these factors. The traditional assay using short conserved regions may not be accurate or credible. These findings have important implications with regard to animal husbandry practices that may inadvertently enhance the contact of human and animal bacterial pathogens. PMID- 21625461 TI - Molecular structure of amyloid fibrils controls the relationship between fibrillar size and toxicity. AB - BACKGROUND: According to the prevailing view, soluble oligomers or small fibrillar fragments are considered to be the most toxic species in prion diseases. To test this hypothesis, two conformationally different amyloid states were produced from the same highly pure recombinant full-length prion protein (rPrP). The cytotoxic potential of intact fibrils and fibrillar fragments generated by sonication from these two states was tested using cultured cells. METHODOLOGY/PRINCIPAL FINDINGS: For one amyloid state, fibril fragmentation was found to enhance its cytotoxic potential, whereas for another amyloid state formed within the same amino acid sequence, the fragmented fibrils were found to be substantially less toxic than the intact fibrils. Consistent with the previous studies, the toxic effects were more pronounced for cell cultures expressing normal isoform of the prion protein (PrP(C)) at high levels confirming that cytotoxicity was in part PrP(C)-dependent. Silencing of PrP(C) expression by small hairpin RNAs designed to silence expression of human PrP(C) (shRNA-PrP(C)) diminished the deleterious effects of the two amyloid states to a different extent, suggesting that the role of PrP(C)-mediated and PrP(C)-independent mechanisms depends on the structure of the aggregates. CONCLUSIONS/SIGNIFICANCE: This work provides a direct illustration that the relationship between an amyloid's physical dimension and its toxic potential is not unidirectional but is controlled by the molecular structure of prion protein (PrP) molecules within aggregated states. Depending on the structure, a decrease in size of amyloid fibrils can either enhance or abolish their cytotoxic effect. Regardless of the molecular structure or size of PrP aggregates, silencing of PrP(C) expression can be exploited to reduce their deleterious effects. PMID- 21625462 TI - Whole-genome immunoinformatic analysis of F. tularensis: predicted CTL epitopes clustered in hotspots are prone to elicit a T-cell response. AB - The cellular arm of the immune response plays a central role in the defense against intracellular pathogens, such as F. tularensis. To date, whole genome immunoinformatic analyses were limited either to relatively small genomes (e.g. viral) or to preselected subsets of proteins in complex pathogens. Here we present, for the first time, an unbiased bacterial global immunoinformatic screen of the 1740 proteins of F. tularensis subs. holarctica (LVS), aiming at identification of immunogenic peptides eliciting a CTL response. The very large number of predicted MHC class I binders (about 100,000, IC(50) of 1000 nM or less) required the design of a strategy for further down selection of CTL candidates. The approach developed focused on mapping clusters rich in overlapping predicted epitopes, and ranking these "hotspot" regions according to the density of putative binding epitopes. Limited by the experimental load, we selected to screen a library of 1240 putative MHC binders derived from 104 top ranking highly dense clusters. Peptides were tested for their ability to stimulate IFNgamma secretion from splenocytes isolated from LVS vaccinated C57BL/6 mice. The majority of the clusters contained one or more CTL responder peptides and altogether 127 novel epitopes were identified, of which 82 are non redundant. Accordingly, the level of success in identification of positive CTL responders was 17-25 fold higher than that found for a randomly selected library of 500 predicted MHC binders (IC(50) of 500 nM or less). Most proteins (ca. 2/3) harboring the highly dense hotspots are membrane-associated. The approach for enrichment of true positive CTL epitopes described in this study, which allowed for over 50% increase in the dataset of known T-cell epitopes of F. tularensis, could be applied in immunoinformatic analyses of many other complex pathogen genomes. PMID- 21625463 TI - Expression and purification of recombinant hemoglobin in Escherichia coli. AB - BACKGROUND: Recombinant DNA technologies have played a pivotal role in the elucidation of structure-function relationships in hemoglobin (Hb) and other globin proteins. Here we describe the development of a plasmid expression system to synthesize recombinant Hbs in Escherichia coli, and we describe a protocol for expressing Hbs with low intrinsic solubilities. Since the alpha- and beta-chain Hbs of different species span a broad range of solubilities, experimental protocols that have been optimized for expressing recombinant human HbA may often prove unsuitable for the recombinant expression of wildtype and mutant Hbs of other species. METHODOLOGY/PRINCIPAL FINDINGS: As a test case for our expression system, we produced recombinant Hbs of the deer mouse (Peromyscus maniculatus), a species that has been the subject of research on mechanisms of Hb adaptation to hypoxia. By experimentally assessing the combined effects of induction temperature, induction time and E. coli expression strain on the solubility of recombinant deer mouse Hbs, we identified combinations of expression conditions that greatly enhanced the yield of recombinant protein and which also increased the efficiency of post-translational modifications. CONCLUSION/SIGNIFICANCE: Our protocol should prove useful for the experimental study of recombinant Hbs in many non-human animals. One of the chief advantages of our protocol is that we can express soluble recombinant Hb without co-expressing molecular chaperones, and without the need for additional reconstitution or heme-incorporation steps. Moreover, our plasmid construct contains a combination of unique restriction sites that allows us to produce recombinant Hbs with different alpha- and beta chain subunit combinations by means of cassette mutagenesis. PMID- 21625464 TI - Evaluating nuclei concentration in amyloid fibrillation reactions using back calculation approach. AB - BACKGROUND: In spite of our extensive knowledge of the more than 20 proteins associated with different amyloid diseases, we do not know how amyloid toxicity occurs or how to block its action. Recent contradictory reports suggest that the fibrils and/or the oligomer precursors cause toxicity. An estimate of their temporal concentration may broaden understanding of the amyloid aggregation process. METHODOLOGY/PRINCIPAL FINDINGS: Assuming that conversion of folded protein to fibril is initiated by a nucleation event, we back-calculate the distribution of nuclei concentration. The temporal in vitro concentration of nuclei for the model hormone, recombinant human insulin, is estimated to be in the picomolar range. This is a conservative estimate since the back-calculation method is likely to overestimate the nuclei concentration because it does not take into consideration fibril fragmentation, which would lower the amount of nuclei CONCLUSIONS: Because of their propensity to form aggregates (non-ordered) and fibrils (ordered), this very low concentration could explain the difficulty in isolating and blocking oligomers or nuclei toxicity and the long onset time for amyloid diseases. PMID- 21625465 TI - Changes in parasite virulence induced by the disruption of a single member of the 235 kDa rhoptry protein multigene family of Plasmodium yoelii. AB - Invasion of the erythrocyte by the merozoites of the malaria parasite is a complex process involving a range of receptor-ligand interactions. Two protein families termed Erythrocyte Binding Like (EBL) proteins and Reticulocyte Binding Protein Homologues (RH) play an important role in host cell recognition by the merozoite. In the rodent malaria parasite, Plasmodium yoelii, the 235 kDa rhoptry proteins (Py235) are coded for by a multigene family and are members of the RH. In P. yoelii Py235 as well as a single member of EBL have been shown to be key mediators of virulence enabling the parasite to invade a wider range of host erythrocytes. One member of Py235, PY01365 is most abundantly transcribed in parasite populations and the protein specifically binds to erythrocytes and is recognized by the protective monoclonal antibody 25.77, suggesting a key role of this particular member in virulence. Recent studies have indicated that overall levels of Py235 expression are essential for parasite virulence. Here we show that disruption of PY01365 in the virulent YM line directly impacts parasite virulence. Furthermore the disruption of PY01365 leads to a reduction in the number of schizonts that express members of Py235 that react specifically with the mcAb 25.77. Erythrocyte binding assays show reduced binding of Py235 to red blood cells in the PY01365 knockout parasite as compared to YM. While our results identify PY01365 as a mediator of parasite virulence, they also confirm that other members of Py235 are able to substitute for PY01365. PMID- 21625466 TI - Life-cycle switching and coexistence of species with no niche differentiation. AB - The increasing evidence of coexistence of cryptic species with no recognized niche differentiation has called attention to mechanisms reducing competition that are not based on niche-differentiation. Only sex-based mechanisms have been shown to create the negative feedback needed for stable coexistence of competitors with completely overlapping niches. Here we show that density dependent sexual and diapause investment can mediate coexistence of facultative sexual species having identical niches. We modelled the dynamics of two competing cyclical parthenogens with species-specific density-dependent sexual and diapause investment and either equal or different competitive abilities. We show that investment in sexual reproduction creates an opportunity for other species to invade and become established. This may happen even if the invading species is an inferior competitor. Our results suggests a previously unnoticed mechanism for species coexistence and can be extended to other facultative sexual species and species investing in diapause where similar density-dependent life-history switches could act to promote coexistence. PMID- 21625467 TI - Unfaithful maintenance of methylation imprints due to loss of maternal nuclear Dnmt1 during somatic cell nuclear transfer. AB - The low success rate of somatic cell nuclear transfer (SCNT) in mammalian cloning is largely due to imprinting problems. However, little is known about the mechanisms of reprogramming imprinted genes during SCNT. Parental origin-specific DNA methylation regulates the monoallelic expression of imprinted genes. In natural fertilization, methylation imprints are established in the parental germline and maintained throughout embryonic development. However, it is unclear whether methylation imprints are protected from global changes of DNA methylation in cloned preimplantation embryos. Here, we demonstrate that cloned porcine preimplantation embryos exhibit demethylation at differentially methylated regions (DMRs) of imprinted genes; in particular, demethylation occurs during the first two cell cycles. By RNAi-mediated knockdown, we found that Dnmt1 is required for the maintenance of methylation imprints in porcine preimplantation embryos. However, no clear signals were detected in the nuclei of oocytes and preimplantation embryos by immunofluorescence. Thus, Dnmt1 is present at very low levels in the nuclei of porcine oocytes and preimplantation embryos and maintains methylation imprints. We further showed that methylation imprints were rescued in nonenucleated metaphase II (MII) oocytes. Our results indicate that loss of Dnmt1 in the maternal nucleus during SCNT significantly contributes to the unfaithful maintenance of methylation imprints in cloned embryos. PMID- 21625468 TI - Interaction between axons and specific populations of surrounding cells is indispensable for collateral formation in the mammillary system. AB - BACKGROUND: An essential phenomenon during brain development is the extension of long collateral branches by axons. How the local cellular environment contributes to the initial sprouting of these branches in specific points of an axonal shaft remains unclear. METHODOLOGY/PRINCIPAL FINDINGS: The principal mammillary tract (pm) is a landmark axonal bundle connecting ventral diencephalon to brainstem (through the mammillotegmental tract, mtg). Late in development, the axons of the principal mammillary tract sprout collateral branches at a very specific point forming a large bundle whose target is the thalamus. Inspection of this model showed a number of distinct, identified cell populations originated in the dorsal and the ventral diencephalon and migrating during development to arrange themselves into several discrete groups around the branching point. Further analysis of this system in several mouse lines carrying mutant alleles of genes expressed in defined subpopulations (including Pax6, Foxb1, Lrp6 and Gbx2) together with the use of an unambiguous genetic marker of mammillary axons revealed: 1) a specific group of Pax6-expressing cells in close apposition with the prospective branching point is indispensable to elicit axonal branching in this system; and 2) cooperation of transcription factors Foxb1 and Pax6 to differentially regulate navigation and fasciculation of distinct branches of the principal mammillary tract. CONCLUSIONS/SIGNIFICANCE: Our results define for the first time a model system where interaction of the axonal shaft with a specific group of surrounding cells is essential to promote branching. Additionally, we provide insight on the cooperative transcriptional regulation necessary to promote and organize an intricate axonal tree. PMID- 21625469 TI - Aspartoacylase-lacZ knockin mice: an engineered model of Canavan disease. AB - Canavan Disease (CD) is a recessive leukodystrophy caused by loss of function mutations in the gene encoding aspartoacylase (ASPA), an oligodendrocyte-enriched enzyme that hydrolyses N-acetylaspartate (NAA) to acetate and aspartate. The neurological phenotypes of different rodent models of CD vary considerably. Here we report on a novel targeted aspa mouse mutant expressing the bacterial beta Galactosidase (lacZ) gene under the control of the aspa regulatory elements. X Gal staining in known ASPA expression domains confirms the integrity of the modified locus in heterozygous aspa lacZ-knockin (aspa(lacZ/+)) mice. In addition, abundant ASPA expression was detected in Schwann cells. Homozygous (aspa(lacZ/lacZ)) mutants are ASPA-deficient, show CD-like histopathology and moderate neurological impairment with behavioural deficits that are more pronounced in aspa(lacZ/lacZ) males than females. Non-invasive ultrahigh field proton magnetic resonance spectroscopy revealed increased levels of NAA, myo inositol and taurine in the aspa(lacZ/lacZ) brain. Spongy degeneration was prominent in hippocampus, thalamus, brain stem, and cerebellum, whereas white matter of optic nerve and corpus callosum was spared. Intracellular vacuolisation in astrocytes coincides with axonal swellings in cerebellum and brain stem of aspa(lacZ/lacZ) mutants indicating that astroglia may act as an osmolyte buffer in the aspa-deficient CNS. In summary, the aspa(lacZ) mouse is an accurate model of CD and an important tool to identify novel aspects of its complex pathology. PMID- 21625470 TI - Testing the efficacy of a multi-component DNA-prime/DNA-boost vaccine against Trypanosoma cruzi infection in dogs. AB - BACKGROUND: Trypanosoma cruzi, the etiologic agent of Chagas Disease, is a major vector borne health problem in Latin America and an emerging infectious disease in the United States. METHODS: We tested the efficacy of a multi-component DNA prime/DNA-boost vaccine (TcVac1) against experimental T. cruzi infection in a canine model. Dogs were immunized with antigen-encoding plasmids and cytokine adjuvants, and two weeks after the last immunization, challenged with T. cruzi trypomastigotes. We measured antibody responses by ELISA and haemagglutination assay, parasitemia and infectivity to triatomines by xenodiagnosis, and performed electrocardiography and histology to assess myocardial damage and tissue pathology. RESULTS: Vaccination with TcVac1 elicited parasite-and antigen specific IgM and IgG (IgG2>IgG1) responses. Upon challenge infection, TcVac1 vaccinated dogs, as compared to non-vaccinated controls dogs, responded to T. cruzi with a rapid expansion of antibody response, moderately enhanced CD8(+) T cell proliferation and IFN-gamma production, and suppression of phagocytes' activity evidenced by decreased myeloperoxidase and nitrite levels. Subsequently, vaccinated dogs controlled the acute parasitemia by day 37 pi (44 dpi in non vaccinated dogs), and exhibited a moderate decline in infectivity to triatomines. TcVac1-immunized dogs did not control the myocardial parasite burden and electrocardiographic and histopatholgic cardiac alterations that are the hallmarks of acute Chagas disease. During the chronic stage, TcVac1-vaccinated dogs exhibited a moderate decline in cardiac alterations determined by EKG and anatomo-/histo-pathological analysis while chronically-infected/non-vaccinated dogs continued to exhibit severe EKG alterations. CONCLUSIONS: Overall, these results demonstrated that TcVac1 provided a partial resistance to T. cruzi infection and Chagas disease, and provide an impetus to improve the vaccination strategy against Chagas disease. PMID- 21625471 TI - Revisiting the simplified bernoulli equation. AB - BACKGROUND: The assessment of the severity of aortic valve stenosis is done by either invasive catheterization or non-invasive Doppler Echocardiography in conjunction with the simplified Bernoulli equation. The catheter measurement is generally considered more accurate, but the procedure is also more likely to have dangerous complications. OBJECTIVE: The focus here is on examining computational fluid dynamics as an alternative method for analyzing the echo data and determining whether it can provide results similar to the catheter measurement. METHODS: An in vitro heart model with a rigid orifice is used as a first step in comparing echocardiographic data, which uses the simplified Bernoulli equation, catheterization, and echocardiographic data, which uses computational fluid dynamics (i.e., the Navier-Stokes equations). RESULTS: For a 0.93cm(2) orifice, the maximum pressure gradient predicted by either the simplified Bernoulli equation or computational fluid dynamics was not significantly different from the experimental catheter measurement (p > 0.01). For a smaller 0.52cm(2) orifice, there was a small but significant difference (p < 0.01) between the simplified Bernoulli equation and the computational fluid dynamics simulation, with the computational fluid dynamics simulation giving better agreement with experimental data for some turbulence models. CONCLUSION: For this simplified, in vitro system, the use of computational fluid dynamics provides an improvement over the simplified Bernoulli equation with the biggest improvement being seen at higher valvular stenosis levels. PMID- 21625472 TI - Standard operation procedures for conducting the on-the-road driving test, and measurement of the standard deviation of lateral position (SDLP). AB - This review discusses the methodology of the standardized on-the-road driving test and standard operation procedures to conduct the test and analyze the data. The on-the-road driving test has proven to be a sensitive and reliable method to examine driving ability after administration of central nervous system (CNS) drugs. The test is performed on a public highway in normal traffic. Subjects are instructed to drive with a steady lateral position and constant speed. Its primary parameter, the standard deviation of lateral position (SDLP), ie, an index of 'weaving', is a stable measure of driving performance with high test retest reliability. SDLP differences from placebo are dose-dependent, and do not depend on the subject's baseline driving skills (placebo SDLP). It is important that standard operation procedures are applied to conduct the test and analyze the data in order to allow comparisons between studies from different sites. PMID- 21625473 TI - A novel requirement for Janus kinases as mediators of drug resistance induced by fibroblast growth factor-2 in human cancer cells. AB - The development of resistance to chemotherapy is a major cause of cancer-related death. Elucidating the mechanisms of drug resistance should thus lead to novel therapeutic strategies. Fibroblast growth factor (FGF)-2 signaling induces the assembly of a multi-protein complex that provides tumor cells with the molecular machinery necessary for drug resistance. This complex, which involves protein kinase C (PKC) epsilon, v-raf murine sarcoma viral oncogene homolog B1 (B-RAF) and p70 S6 kinase beta (S6K2), enhances the selective translation of anti apoptotic proteins such as B-cell leukaemia/lymphoma-2 (BCL-2) and inhibitors of apoptosis protein (IAP) family members and these are able to protect multiple cancer cell types from chemotherapy-induced cell death. The Janus kinases (JAKs) are most noted for their critical roles in mediating cytokine signaling and immune responses. Here, we show that JAKs have novel functions that support their consideration as new targets in therapies aimed at reducing drug resistance. As an example, we show that the Janus kinase TYK2 is phosphorylated downstream of FGF-2 signaling and required for the full phosphorylation of extracellular signal regulated kinase (ERK) 1/2. Moreover, TYK2 is necessary for the induction of key anti-apoptotic proteins, such as BCL-2 and myeloid cell leukemia sequence (MCL) 1, and for the promotion of cell survival upon FGF-2. Silencing JAK1, JAK2 or TYK2 using RNA interference (RNAi) inhibits FGF2-mediated proliferation and results in the sensitization of tumor cells to chemotherapy-induced killing. These effects are independent of activation of signal transducer and activator of transcription (STAT) 1, STAT3 and STAT5A/B, the normal targets of JAK signaling. Instead, TYK2 associates with the other kinases previously implicated in FGF-2 mediated drug resistance. In light of these findings we hypothesize that TYK2 and other JAKs are important modulators of FGF-2-driven cell survival and that inhibitors of these kinases will likely improve the effectiveness of other cancer therapies. PMID- 21625474 TI - Visual genome-wide RNAi screening to identify human host factors required for Trypanosoma cruzi infection. AB - The protozoan parasite Trypanosoma cruzi is the etiologic agent of Chagas disease, a neglected tropical infection that affects millions of people in the Americas. Current chemotherapy relies on only two drugs that have limited efficacy and considerable side effects. Therefore, the development of new and more effective drugs is of paramount importance. Although some host cellular factors that play a role in T. cruzi infection have been uncovered, the molecular requirements for intracellular parasite growth and persistence are still not well understood. To further study these host-parasite interactions and identify human host factors required for T. cruzi infection, we performed a genome-wide RNAi screen using cellular microarrays of a printed siRNA library that spanned the whole human genome. The screening was reproduced 6 times and a customized algorithm was used to select as hits those genes whose silencing visually impaired parasite infection. The 162 strongest hits were subjected to a secondary screening and subsequently validated in two different cell lines. Among the fourteen hits confirmed, we recognized some cellular membrane proteins that might function as cell receptors for parasite entry and others that may be related to calcium release triggered by parasites during cell invasion. In addition, two of the hits are related to the TGF-beta signaling pathway, whose inhibition is already known to diminish levels of T. cruzi infection. This study represents a significant step toward unveiling the key molecular requirements for host cell invasion and revealing new potential targets for antiparasitic therapy. PMID- 21625475 TI - Elevated serum uric acid is associated with high circulating inflammatory cytokines in the population-based Colaus study. AB - BACKGROUND: The relation of serum uric acid (SUA) with systemic inflammation has been little explored in humans and results have been inconsistent. We analyzed the association between SUA and circulating levels of interleukin-6 (IL-6), interleukin-1beta (IL-1beta), tumor necrosis factor- alpha (TNF-alpha) and C reactive protein (CRP). METHODS AND FINDINGS: This cross-sectional population based study conducted in Lausanne, Switzerland, included 6085 participants aged 35 to 75 years. SUA was measured using uricase-PAP method. Plasma TNF-alpha, IL 1beta and IL-6 were measured by a multiplexed particle-based flow cytometric assay and hs-CRP by an immunometric assay. The median levels of SUA, IL-6, TNF alpha, CRP and IL-1beta were 355 umol/L, 1.46 pg/mL, 3.04 pg/mL, 1.2 mg/L and 0.34 pg/mL in men and 262 umol/L, 1.21 pg/mL, 2.74 pg/mL, 1.3 mg/L and 0.45 pg/mL in women, respectively. SUA correlated positively with IL-6, TNF-alpha and CRP and negatively with IL-1beta (Spearman r: 0.04, 0.07, 0.20 and 0.05 in men, and 0.09, 0.13, 0.30 and 0.07 in women, respectively, P<0.05). In multivariable analyses, SUA was associated positively with CRP (beta coefficient +/- SE = 0.35+/-0.02, P<0.001), TNF-alpha (0.08+/-0.02, P<0.001) and IL-6 (0.10+/-0.03, P<0.001), and negatively with IL-1beta (-0.07+/-0.03, P = 0.027). Upon further adjustment for body mass index, these associations were substantially attenuated. CONCLUSIONS: SUA was associated positively with IL-6, CRP and TNF-alpha and negatively with IL-1beta, particularly in women. These results suggest that uric acid contributes to systemic inflammation in humans and are in line with experimental data showing that uric acid triggers sterile inflammation. PMID- 21625476 TI - TRAIL-R4 promotes tumor growth and resistance to apoptosis in cervical carcinoma HeLa cells through AKT. AB - BACKGROUND: TRAIL/Apo2L is a pro-apoptotic ligand of the TNF family that engages the apoptotic machinery through two pro-apoptotic receptors, TRAIL-R1 and TRAIL R2. This cell death program is tightly controlled by two antagonistic receptors, TRAIL-R3 and TRAIL-R4, both devoid of a functional death domain, an intracellular region of the receptor, required for the recruitment and the activation of initiator caspases. Upon TRAIL-binding, TRAIL-R4 forms a heteromeric complex with the agonistic receptor TRAIL-R2 leading to reduced caspase-8 activation and apoptosis. METHODOLOGY/PRINCIPAL FINDINGS: We provide evidence that TRAIL-R4 can also exhibit, in a ligand independent manner, signaling properties in the cervical carcinoma cell line HeLa, through Akt. Ectopic expression of TRAIL-R4 in HeLa cells induced morphological changes, with cell rounding, loss of adherence and markedly enhanced cell proliferation in vitro and tumor growth in vivo. Disruption of the PI3K/Akt pathway using the pharmacological inhibitor LY294002, siRNA targeting the p85 regulatory subunit of phosphatidylinositol-3 kinase, or by PTEN over-expression, partially restored TRAIL-mediated apoptosis in these cells. Moreover, the Akt inhibitor, LY294002, restituted normal cell proliferation index in HeLa cells expressing TRAIL-R4. CONCLUSIONS/SIGNIFICANCE: Altogether, these results indicate that, besides its ability to directly inhibit TRAIL-induced cell death at the membrane, TRAIL-R4 can also trigger the activation of signaling pathways leading to cell survival and proliferation in HeLa cells. Our findings raise the possibility that TRAIL-R4 may contribute to cervical carcinogenesis. PMID- 21625477 TI - Cumulative viral load and virologic decay patterns after antiretroviral therapy in HIV-infected subjects influence CD4 recovery and AIDS. AB - BACKGROUND: The impact of viral load (VL) decay and cumulative VL on CD4 recovery and AIDS after highly-active antiretroviral therapy (HAART) is unknown. METHODS AND FINDINGS: Three virologic kinetic parameters (first year and overall exponential VL decay constants, and first year VL slope) and cumulative VL during HAART were estimated for 2,278 patients who initiated HAART in the U.S. Military HIV Natural History Study. CD4 and VL trajectories were computed using linear and nonlinear Generalized Estimating Equations models. Multivariate Poisson and linear regression models were used to determine associations of VL parameters with CD4 recovery, adjusted for factors known to correlate with immune recovery. Cumulative VL higher than the sample median was independently associated with an increased risk of AIDS (relative risk 2.38, 95% confidence interval 1.56-3.62, p<0.001). Among patients with VL suppression, first year VL decay and slope were independent predictors of early CD4 recovery (p = 0.001) and overall gain (p<0.05). Despite VL suppression, those with slow decay during the first year of HAART as well as during the entire therapy period (overall), in general, gained less CD4 cells compared to the other subjects (133 vs. 195.4 cells/uL; p = 0.001) even after adjusting for potential confounders. CONCLUSIONS: In a cohort with free access to healthcare, independent of established predictors of AIDS and CD4 recovery during HAART, cumulative VL and virologic decay patterns were associated with AIDS and distinct aspects of CD4 reconstitution. PMID- 21625478 TI - Anti-influenza activity of marchantins, macrocyclic bisbibenzyls contained in liverworts. AB - The H1N1 influenza A virus of swine-origin caused pandemics throughout the world in 2009 and the highly pathogenic H5N1 avian influenza virus has also caused epidemics in Southeast Asia in recent years. The threat of influenza A thus remains a serious global health issue and novel drugs that target these viruses are highly desirable. Influenza A possesses an endonuclease within its RNA polymerase which comprises PA, PB1 and PB2 subunits. To identify potential new anti-influenza compounds in our current study, we screened 33 different types of phytochemicals using a PA endonuclease inhibition assay in vitro and an anti influenza A virus assay. The marchantins are macrocyclic bisbibenzyls found in liverworts, and plagiochin A and perrottetin F are marchantin-related phytochemicals. We found from our screen that marchantin A, B, E, plagiochin A and perrottetin F inhibit influenza PA endonuclease activity in vitro. These compounds have a 3,4-dihydroxyphenethyl group in common, indicating the importance of this moiety for the inhibition of PA endonuclease. Docking simulations of marchantin E with PA endonuclease suggest a putative "fitting and chelating model" as the mechanism underlying PA endonuclease inhibition. The docking amino acids are well conserved between influenza A and B. In a cultured cell system, marchantin E was further found to inhibit the growth of both H3N2 and H1N1 influenza A viruses, and marchantin A, E and perrotein F showed inhibitory properties towards the growth of influenza B. These marchantins also decreased the viral infectivity titer, with marchantin E showing the strongest activity in this assay. We additionally identified a chemical group that is conserved among different anti-influenza chemicals including marchantins, green tea catechins and dihydroxy phenethylphenylphthalimides. Our present results indicate that marchantins are candidate anti-influenza drugs and demonstrate the utility of the PA endonuclease assay in the screening of phytochemicals for anti influenza characteristics. PMID- 21625479 TI - The role of perfusion computed tomography in the prediction of cerebral hyperperfusion syndrome. AB - BACKGROUND: Hyperperfusion syndrome (HPS) following carotid angioplasty with stenting (CAS) is associated with significant morbidity and mortality. At present, there are no reliable parameters to predict HPS. The aim of this study was to clarify whether perfusion computed tomography (CT) is a feasible and reliable tool in predicting HPS after CAS. METHODOLOGY/PRINCIPAL FINDINGS: We performed a retrospective case-control study of 54 patients (11 HPS patients and 43 non-HPS) with unilateral severe stenosis of the carotid artery who underwent CAS. We compared the prevalence of vascular risk factors and perfusion CT parameters including regional cerebral blood volume (rCBV), regional cerebral blood flow (rCBF), and time to peak (TTP) within seven days prior to CAS. Demographic information, risk factors for atherosclerosis, and perfusion CT parameters were evaluated by multivariable logistic regression analysis. The rCBV index was calculated as [(ipsilateral rCBV - contralateral rCBV)/contralateral rCBV], and indices of rCBF and TTP were similarly calculated. We found that eleven patients had HPS, including five with intracranial hemorrhages (ICHs) of whom three died. After a comparison with non-HPS control subjects, independent predictors of HPS included the severity of ipsilateral carotid artery stenosis, 3 hour mean systolic blood pressure (3 h SBP) after CAS, pre-stenting rCBV index >0.15 and TTP index >0.22. CONCLUSIONS/SIGNIFICANCE: The combination of severe ipsilateral carotid stenosis, 3 h SBP after CAS, rCBV index and TTP index provides a potential screening tool for predicting HPS in patients with unilateral carotid stenosis receiving CAS. In addition, adequate management of post-stenting blood pressure is the most important treatable factor in preventing HPS in these high risk patients. PMID- 21625480 TI - Emergence of epidemic Neisseria meningitidis serogroup X meningitis in Togo and Burkina Faso. AB - Serogroup X meningococci (NmX) historically have caused sporadic and clustered meningitis cases in sub-Saharan Africa. To study recent NmX epidemiology, we analyzed data from population-based, sentinel and passive surveillance, and outbreak investigations of bacterial meningitis in Togo and Burkina Faso during 2006-2010. Cerebrospinal fluid specimens were analyzed by PCR. In Togo during 2006-2009, NmX accounted for 16% of the 702 confirmed bacterial meningitis cases. Kozah district experienced an NmX outbreak in March 2007 with an NmX seasonal cumulative incidence of 33/100,000. In Burkina Faso during 2007-2010, NmX accounted for 7% of the 778 confirmed bacterial meningitis cases, with an increase from 2009 to 2010 (4% to 35% of all confirmed cases, respectively). In 2010, NmX epidemics occurred in northern and central regions of Burkina Faso; the highest district cumulative incidence of NmX was estimated as 130/100,000 during March-April. Although limited to a few districts, we have documented NmX meningitis epidemics occurring with a seasonal incidence previously only reported in the meningitis belt for NmW135 and NmA, which argues for development of an NmX vaccine. PMID- 21625481 TI - Peripheral, central and behavioral responses to the cuticular pheromone bouquet in Drosophila melanogaster males. AB - Pheromonal communication is crucial with regard to mate choice in many animals including insects. Drosophila melanogaster flies produce a pheromonal bouquet with many cuticular hydrocarbons some of which diverge between the sexes and differently affect male courtship behavior. Cuticular pheromones have a relatively high weight and are thought to be -- mostly but not only -- detected by gustatory contact. However, the response of the peripheral and central gustatory systems to these substances remains poorly explored. We measured the effect induced by pheromonal cuticular mixtures on (i) the electrophysiological response of peripheral gustatory receptor neurons, (ii) the calcium variation in brain centers receiving these gustatory inputs and (iii) the behavioral reaction induced in control males and in mutant desat1 males, which show abnormal pheromone production and perception. While male and female pheromones induced inhibitory-like effects on taste receptor neurons, the contact of male pheromones on male fore-tarsi elicits a long-lasting response of higher intensity in the dedicated gustatory brain center. We found that the behavior of control males was more strongly inhibited by male pheromones than by female pheromones, but this difference disappeared in anosmic males. Mutant desat1 males showed an increased sensitivity of their peripheral gustatory neurons to contact pheromones and a behavioral incapacity to discriminate sex pheromones. Together our data indicate that cuticular hydrocarbons induce long-lasting inhibitory effects on the relevant taste pathway which may interact with the olfactory pathway to modulate pheromonal perception. PMID- 21625482 TI - Human wavelength discrimination of monochromatic light explained by optimal wavelength decoding of light of unknown intensity. AB - We show that human ability to discriminate the wavelength of monochromatic light can be understood as maximum likelihood decoding of the cone absorptions, with a signal processing efficiency that is independent of the wavelength. This work is built on the framework of ideal observer analysis of visual discrimination used in many previous works. A distinctive aspect of our work is that we highlight a perceptual confound that observers should confuse a change in input light wavelength with a change in input intensity. Hence a simple ideal observer model which assumes that an observer has a full knowledge of input intensity should over-estimate human ability in discriminating wavelengths of two inputs of unequal intensity. This confound also makes it difficult to consistently measure human ability in wavelength discrimination by asking observers to distinguish two input colors while matching their brightness. We argue that the best experimental method for reliable measurement of discrimination thresholds is the one of Pokorny and Smith, in which observers only need to distinguish two inputs, regardless of whether they differ in hue or brightness. We mathematically formulate wavelength discrimination under this wavelength-intensity confound and show a good agreement between our theoretical prediction and the behavioral data. Our analysis explains why the discrimination threshold varies with the input wavelength, and shows how sensitively the threshold depends on the relative densities of the three types of cones in the retina (and in particular predict discriminations in dichromats). Our mathematical formulation and solution can be applied to general problems of sensory discrimination when there is a perceptual confound from other sensory feature dimensions. PMID- 21625483 TI - DNA dynamics is likely to be a factor in the genomic nucleotide repeats expansions related to diseases. AB - Trinucleotide repeats sequences (TRS) represent a common type of genomic DNA motif whose expansion is associated with a large number of human diseases. The driving molecular mechanisms of the TRS ongoing dynamic expansion across generations and within tissues and its influence on genomic DNA functions are not well understood. Here we report results for a novel and notable collective breathing behavior of genomic DNA of tandem TRS, leading to propensity for large local DNA transient openings at physiological temperature. Our Langevin molecular dynamics (LMD) and Markov Chain Monte Carlo (MCMC) simulations demonstrate that the patterns of openings of various TRSs depend specifically on their length. The collective propensity for DNA strand separation of repeated sequences serves as a precursor for outsized intermediate bubble states independently of the G/C content. We report that repeats have the potential to interfere with the binding of transcription factors to their consensus sequence by altered DNA breathing dynamics in proximity of the binding sites. These observations might influence ongoing attempts to use LMD and MCMC simulations for TRS-related modeling of genomic DNA functionality in elucidating the common denominators of the dynamic TRS expansion mutation with potential therapeutic applications. PMID- 21625485 TI - The organophosphate Chlorpyrifos interferes with the responses to 17beta estradiol in the digestive gland of the marine mussel Mytilus galloprovincialis. AB - BACKGROUND: Many pesticides have been shown to act as endocrine disrupters. Although the potencies of currently used pesticides as hormone agonists/antagonists are low compared with those of natural ligands, their ability to act via multiple mechanisms might enhance the biological effect. The organophosphate Chlorpyrifos (CHP) has been shown to be weakly estrogenic and cause adverse neurodevelopmental effects in mammals. However, no information is available on the endocrine effects of CHP in aquatic organisms. In the digestive gland of the bivalve Mytilus galloprovincialis, a target tissue of both estrogens and pesticides, the possible effects of CHP on the responses to the natural estrogen 17beta-estradiol (E(2)) were investigated. METHODOLOGY/PRINCIPAL FINDINGS: Mussels were exposed to CHP (4.5 mg/l, 72 hrs) and subsequently injected with E(2) (6.75 ng/g dw). Responses were evaluated in CHP, E(2) and CHP/E(2) treatment groups at 24 h p.i. by a biomarker/transcriptomic approach. CHP and E(2) induced additive, synergistic, and antagonistic effects on lysosomal biomarkers (lysosomal membrane stability, lysosome/cytoplasm volume ratio, lipofuscin and neutral lipid accumulation). Additive and synergistic effects were also observed on the expression of estrogen-responsive genes (GSTpi, catalase, 5 HTR) evaluated by RT-Q-PCR. The use of a 1.7K cDNA Mytilus microarray showed that CHP, E(2) and CHP/E(2), induced 81, 44, and 65 Differentially Expressed Genes (DEGs), respectively. 24 genes were exclusively shared between CHP and CHP/E(2), only 2 genes between E(2) and CHP/E(2). Moreover, 36 genes were uniquely modulated by CHP/E(2). Gene ontology annotation was used to elucidate the putative mechanisms involved in the responses elicited by different treatments. CONCLUSIONS: The results show complex interactions between CHP and E(2) in the digestive gland, indicating that the combination of certain pesticides and hormones may give rise to unexpected effects at the molecular/cellular level. Overall, these data demonstrate that CHP can interfere with the mussel responses to natural estrogens. PMID- 21625484 TI - A comprehensive evaluation of potential lung function associated genes in the SpiroMeta general population sample. AB - RATIONALE: Lung function measures are heritable traits that predict population morbidity and mortality and are essential for the diagnosis of chronic obstructive pulmonary disease (COPD). Variations in many genes have been reported to affect these traits, but attempts at replication have provided conflicting results. Recently, we undertook a meta-analysis of Genome Wide Association Study (GWAS) results for lung function measures in 20,288 individuals from the general population (the SpiroMeta consortium). OBJECTIVES: To comprehensively analyse previously reported genetic associations with lung function measures, and to investigate whether single nucleotide polymorphisms (SNPs) in these genomic regions are associated with lung function in a large population sample. METHODS: We analysed association for SNPs tagging 130 genes and 48 intergenic regions (+/ 10 kb), after conducting a systematic review of the literature in the PubMed database for genetic association studies reporting lung function associations. RESULTS: The analysis included 16,936 genotyped and imputed SNPs. No loci showed overall significant association for FEV(1) or FEV(1)/FVC traits using a carefully defined significance threshold of 1.3*10(-5). The most significant loci associated with FEV(1) include SNPs tagging MACROD2 (P = 6.81*10(-5)), CNTN5 (P = 4.37*10(-4)), and TRPV4 (P = 1.58*10(-3)). Among ever-smokers, SERPINA1 showed the most significant association with FEV(1) (P = 8.41*10(-5)), followed by PDE4D (P = 1.22*10(-4)). The strongest association with FEV(1)/FVC ratio was observed with ABCC1 (P = 4.38*10(-4)), and ESR1 (P = 5.42*10(-4)) among ever-smokers. CONCLUSIONS: Polymorphisms spanning previously associated lung function genes did not show strong evidence for association with lung function measures in the SpiroMeta consortium population. Common SERPINA1 polymorphisms may affect FEV(1) among smokers in the general population. PMID- 21625486 TI - Intranasal immunization with recombinant HA and mast cell activator C48/80 elicits protective immunity against 2009 pandemic H1N1 influenza in mice. AB - BACKGROUND: Pandemic influenza represents a major threat to global health. Vaccination is the most economic and effective strategy to control influenza pandemic. Conventional vaccine approach, despite being effective, has a number of major deficiencies including limited range of protection, total dependence on embryonated eggs for production, and time consuming for vaccine production. There is an urgent need to develop novel vaccine strategies to overcome these deficiencies. METHODOLOGY/PRINCIPAL FINDINGS: The major objective of this work was to develop a novel vaccine strategy combining recombinant haemagglutinin (HA) protein and a master cell (MC) activator C48/80 for intranasal immunization. We demonstrated in BALB/c mice that MC activator C48/80 had strong adjuvant activity when co-administered with recombinant HA protein intranasally. Vaccination with C48/80 significantly increased the serum IgG and mucosal surface IgA antibody responses against HA protein. Such increases correlated with stronger and durable neutralizing antibody activities, offering protection to vaccinated animals from disease progression after challenge with lethal dose of A/California/04/2009 live virus. Furthermore, protected animals demonstrated significant reduction in lung virus titers, minimal structural alteration in lung tissues as well as higher and balanced production of Th1 and Th2 cytokines in the stimulated splenocytes when compared to those without C48/80. CONCLUSIONS/SIGNIFICANCE: The present study demonstrates that the novel vaccine approach of combining recombinant HA and mucosal adjuvant C48/80 is safe and effective in eliciting protective immunity in mice. Future studies on the mechanism of action of C48/80 and potential combination with other vaccine strategies such as prime and boost approach may help to induce even more potent and broad immune responses against viruses from various clades. PMID- 21625487 TI - Unexpected course of nonlinear cardiac interbeat interval dynamics during childhood and adolescence. AB - The fluctuations of the cardiac interbeat series contain rich information because they reflect variations of other functions on different time scales (e.g., respiration or blood pressure control). Nonlinear measures such as complexity and fractal scaling properties derived from 24 h heart rate dynamics of healthy subjects vary from childhood to old age. In this study, the age-related variations during childhood and adolescence were addressed. In particular, the cardiac interbeat interval series was quantified with respect to complexity and fractal scaling properties. The R-R interval series of 409 healthy children and adolescents (age range: 1 to 22 years, 220 females) was analyzed with respect to complexity (Approximate Entropy, ApEn) and fractal scaling properties on three time scales: long-term (slope beta of the power spectrum, log power vs. log frequency, in the frequency range 10(-4) to 10(-2) Hz) intermediate-term (DFA, detrended fluctuation analysis, alpha(2)) and short-term (DFA alpha(1)). Unexpectedly, during age 7 to 13 years beta and ApEn were higher compared to the age <7 years and age >13 years (beta: -1.06 vs. -1.21; ApEn: 0.88 vs. 0.74). Hence, the heart rate dynamics were closer to a 1/f power law and most complex between 7 and 13 years. However, DFA alpha(1) and alpha(2) increased with progressing age similar to measures reflecting linear properties. In conclusion, the course of long-term fractal scaling properties and complexity of heart rate dynamics during childhood and adolescence indicates that these measures reflect complex changes possibly linked to hormonal changes during pre-puberty and puberty. PMID- 21625488 TI - Potent antioxidant and genoprotective effects of boeravinone G, a rotenoid isolated from Boerhaavia diffusa. AB - BACKGROUND AND AIMS: Free radicals are implicated in the aetiology of some gastrointestinal disorders such as gastric ulcer, colorectal cancer and inflammatory bowel disease. In the present study we investigated the antioxidant and genoprotective activity of some rotenoids (i.e. boeravinones) isolated from the roots of Boerhaavia diffusa, a plant used in the Ayurvedic medicine for the treatment of diseases affecting the gastrointestinal tract. METHODS/PRINCIPAL FINDINGS: Antioxidant activity has been evaluated using both chemical (Electron Spin Resonance spectroscopy, ESR) and Caco-2 cells-based (TBARS and ROS) assays. DNA damage was evaluated by Comet assay, while pERK(1/2) and phospho-NF-kB p65 levels were estimated by western blot. Boeravinones G, D and H significantly reduced the signal intensity of ESR induced by hydroxyl radicals, suggesting a scavenging activity. Among rotenoids tested, boeravinone G exerted the most potent effect. Boeravinone G inhibited both TBARS and ROS formation induced by Fenton's reagent, increased SOD activity and reduced H(2)O(2)-induced DNA damage. Finally, boeravinone G reduced the levels of pERK(1) and phospho-NF-kB p65 (but not of pERK(2)) increased by Fenton's reagent. CONCLUSIONS: It is concluded that boeravinone G exhibits an extraordinary potent antioxidant activity (significant effect in the nanomolar range). The MAP kinase and NF-kB pathways seem to be involved in the antioxidant effect of boeravinone G. Boeravinone G might be considered as lead compound for the development of drugs potentially useful against those pathologies whose aetiology is related to ROS-mediated injuries. PMID- 21625489 TI - Cost-effectiveness of HIV screening in STD clinics, emergency departments, and inpatient units: a model-based analysis. AB - BACKGROUND: Identifying and treating persons with human immunodeficiency virus (HIV) infection early in their disease stage is considered an effective means of reducing the impact of the disease. We compared the cost-effectiveness of HIV screening in three settings, sexually transmitted disease (STD) clinics serving men who have sex with men, hospital emergency departments (EDs), settings where patients are likely to be diagnosed early, and inpatient diagnosis based on clinical manifestations. METHODS AND FINDINGS: We developed the Progression and Transmission of HIV/AIDS model, a health state transition model that tracks index patients and their infected partners from HIV infection to death. We used program characteristics for each setting to compare the incremental cost per quality adjusted life year gained from early versus late diagnosis and treatment. We ran the model for 10,000 index patients for each setting, examining alternative scenarios, excluding and including transmission to partners, and assuming HAART was initiated at a CD4 count of either 350 or 500 cells/uL. Screening in STD clinics and EDs was cost-effective compared with diagnosing inpatients, even when including only the benefits to the index patients. Screening patients in STD clinics, who have less-advanced disease, was cost-effective compared with ED screening when treatment with HAART was initiated at a CD4 count of 500 cells/uL. When the benefits of reduced transmission to partners from early diagnosis were included, screening in settings with less-advanced disease stages was cost-saving compared with screening later in the course of infection. The study was limited by a small number of observations on CD4 count at diagnosis and by including transmission only to first generation partners of the index patients. CONCLUSIONS: HIV prevention efforts can be advanced by screening in settings where patients present with less-advanced stages of HIV infection and by initiating treatment with HAART earlier in the course of infection. PMID- 21625490 TI - Genome-wide association study for atopy and allergic rhinitis in a Singapore Chinese population. AB - Allergic rhinitis (AR) is an atopic disease which affects about 600 million people worldwide and results from a complex interplay between genetic and environmental factors. However genetic association studies on known candidate genes yielded variable results. The aim of this study is to identify the genetic variants that influence predisposition towards allergic rhinitis in an ethnic Chinese population in Singapore using a genome-wide association study (GWAS) approach. A total of 4461 ethnic Chinese volunteers were recruited in Singapore and classified according to their allergic disease status. The GWAS included a discovery stage comparing 515 atopic cases (including 456 AR cases) and 486 non allergic non-rhinitis (NANR) controls. The top SNPs were then validated in a replication cohort consisting of a separate 2323 atopic cases (including 676 AR cases) and 511 NANR controls. Two SNPs showed consistent association in both discovery and replication phases; MRPL4 SNP rs8111930 on 19q13.2 (OR = 0.69, P(combined) = 4.46*10(-05)) and BCAP SNP rs505010 on chromosome 10q24.1 (OR = 0.64, P(combined) = 1.10*10(-04)). In addition, we also replicated multiple associations within known candidates regions such as HLA-DQ and NPSR1 locus in the discovery phase. Our study suggests that MRPL4 and BCAP, key components of the HIF-1alpha and PI3K/Akt signaling pathways respectively, are two novel candidate genes for atopy and allergic rhinitis. Further study on these molecules and their signaling pathways would help in understanding of the pathogenesis of allergic rhinitis and identification of targets for new therapeutic intervention. PMID- 21625491 TI - Adipose inflammation initiates recruitment of leukocytes to mouse femoral artery: role of adipo-vascular axis in chronic inflammation. AB - BACKGROUND: Although inflammation within adipose tissues is known to play a role in metabolic syndrome, the causative connection between inflamed adipose tissue and atherosclerosis is not fully understood. In the present study, we examined the direct effects of adipose tissue on macro-vascular inflammation using intravital microscopic analysis of the femoral artery after adipose tissue transplantation. METHODS AND RESULTS: We obtained subcutaneous (SQ) and visceral (VIS) adipose tissues from C57BL/6 mice fed normal chow (NC) or a high fat diet (HF), then transplanted the tissues into the perivascular area of the femoral artery of recipient C57/BL6 mice. Quantitative intravital microscopic analysis revealed an increase in adherent leukocytes after adipose tissue transplantation, with VIS found to induce significantly more leukocyte accumulation as compared to SQ. Moreover, adipose tissues from HF fed mice showed significantly more adhesion to the femoral artery. Simultaneous flow cytometry demonstrated upregulation of CD11b on peripheral granulocyte and monocytes after adipose tissue transplantation. We also observed dominant expressions of the inflammatory cytokine IL-6, and chemokines MCP-1 and MIP-1beta in the stromal vascular fraction (SVF) of these adipose tissues as well as sera of recipient mice after transplantation. Finally, massive accumulations of pro-inflammatory and dendritic cells were detected in mice with VIS transplantation as compared to SQ, as well as in HF mice as compared to those fed NC. CONCLUSION: Our in vivo findings indicate that adipose tissue stimulates leukocyte accumulation in the femoral artery. The underlying mechanisms involve upregulation of CD11b in leukocytes, induction of cytokines and chemokines, and accumulation of pro-inflammatory cells in the SVF. PMID- 21625492 TI - Perception of biological motion in schizophrenia and healthy individuals: a behavioral and FMRI study. AB - BACKGROUND: Anomalous visual perception is a common feature of schizophrenia plausibly associated with impaired social cognition that, in turn, could affect social behavior. Past research suggests impairment in biological motion perception in schizophrenia. Behavioral and functional magnetic resonance imaging (fMRI) experiments were conducted to verify the existence of this impairment, to clarify its perceptual basis, and to identify accompanying neural concomitants of those deficits. METHODOLOGY/FINDINGS: In Experiment 1, we measured ability to detect biological motion portrayed by point-light animations embedded within masking noise. Experiment 2 measured discrimination accuracy for pairs of point light biological motion sequences differing in the degree of perturbation of the kinematics portrayed in those sequences. Experiment 3 measured BOLD signals using event-related fMRI during a biological motion categorization task. Compared to healthy individuals, schizophrenia patients performed significantly worse on both the detection (Experiment 1) and discrimination (Experiment 2) tasks. Consistent with the behavioral results, the fMRI study revealed that healthy individuals exhibited strong activation to biological motion, but not to scrambled motion in the posterior portion of the superior temporal sulcus (STSp). Interestingly, strong STSp activation was also observed for scrambled or partially scrambled motion when the healthy participants perceived it as normal biological motion. On the other hand, STSp activation in schizophrenia patients was not selective to biological or scrambled motion. CONCLUSION: Schizophrenia is accompanied by difficulties discriminating biological from non-biological motion, and associated with those difficulties are altered patterns of neural responses within brain area STSp. The perceptual deficits exhibited by schizophrenia patients may be an exaggerated manifestation of neural events within STSp associated with perceptual errors made by healthy observers on these same tasks. The present findings fit within the context of theories of delusion involving perceptual and cognitive processes. PMID- 21625493 TI - Receptor sorting within endosomal trafficking pathway is facilitated by dynamic actin filaments. AB - Early endosomes (EEs) are known to be a sorting station for internalized molecules destined for degradation, recycling, or other intracellular organelles. Segregation is an essential step in such sorting, but the molecular mechanism of this process remains to be elucidated. Here, we show that actin is required for efficient recycling and endosomal maturation by producing a motile force. Perturbation of actin dynamics by drugs induced a few enlarged EEs containing several degradative vacuoles and also interfered with their transporting ability. Actin repolymerization induced by washout of the drug caused the vacuoles to dissociate and individually translocate toward the perinuclear region. We further elucidated that cortactin, an actin-nucleating factor, was required for transporting contents from within EEs. Actin filaments regulated by cortactin may provide a motile force for efficient sorting within early endosomes. These data suggest that actin filaments coordinate with microtubules to mediate segregation in EEs. PMID- 21625494 TI - Drosophila sperm swim backwards in the female reproductive tract and are activated via TRPP2 ion channels. AB - BACKGROUND: Sperm have but one purpose, to fertilize an egg. In various species including Drosophila melanogaster female sperm storage is a necessary step in the reproductive process. Amo is a homolog of the human transient receptor potential channel TRPP2 (also known as PKD2), which is mutated in autosomal dominant polycystic kidney disease. In flies Amo is required for sperm storage. Drosophila males with Amo mutations produce motile sperm that are transferred to the uterus but they do not reach the female storage organs. Therefore Amo appears to be a mediator of directed sperm motility in the female reproductive tract but the underlying mechanism is unknown. METHODOLOGY/PRINCIPAL FINDINGS: Amo exhibits a unique expression pattern during spermatogenesis. In spermatocytes, Amo is restricted to the endoplasmic reticulum (ER) whereas in mature sperm, Amo clusters at the distal tip of the sperm tail. Here we show that flagellar localization of Amo is required for sperm storage. This raised the question of how Amo at the rear end of sperm regulates forward movement into the storage organs. In order to address this question, we used in vivo imaging of dual labelled sperm to demonstrate that Drosophila sperm navigate backwards in the female reproductive tract. In addition, we show that sperm exhibit hyperactivation upon transfer to the uterus. Amo mutant sperm remain capable of reverse motility but fail to display hyperactivation and directed movement, suggesting that these functions are required for sperm storage in flies. CONCLUSIONS/SIGNIFICANCE: Amo is part of a signalling complex at the leading edge of the sperm tail that modulates flagellar beating and that guides a backwards path into the storage organs. Our data support an evolutionarily conserved role for TRPP2 channels in cilia. PMID- 21625495 TI - Molecular characterizations of a novel putative DNA-binding protein LvDBP23 in marine shrimp L. vannamei tissues and molting stages. AB - BACKGROUND: Litopenaeus Vannamei, well known as pacific white shrimp, is the most popular shrimp in the world shrimp market. Identification and characterization of shrimp muscle regulatory genes are not only important for shrimp genetic improvement, but also facilitate comparative genomic tools for understanding of muscle development and regeneration. METHODOLOGY/PRINCIPAL FINDINGS: A novel mRNA encoding for a putative DNA-binding protein LvDBP23 was identified from Litopenaeus vannamei abdominal muscle cDNA library. The LvDBP23 cDNA contains 639 nucleotides of protein-coding sequence with deduced 212 amino acids of predicted molecular mass 23.32 kDa with glycine-rich domain at amino acid position 94-130. The mRNA sequence is successfully used for producing LvDBP23 recombinant protein in sf9 insect cell expression system. The expression of LvDBP23 mRNA is presented in abdominal muscle and swimming leg muscle, as well as other tissues including intestine, lymphoid and gill. The mRNA expression has the highest level in abdominal muscle in all tested tissues. LVDBP23 transcript during the molt cycle is highly expressed in the intermolt stage. In vitro nucleic acid-binding assays reveal that LvDBP23 protein can bind to both ssDNA and dsDNA, indicating its possible role of regulation of gene transcription. CONCLUSIONS/SIGNIFICANCE: We are the first to report a DNA-binding protein identified from the abdominal muscle tissue of marine shrimp L. Vannamei. Its high-level specific expression during the intermot stage suggests its role in the regulation of muscle buildup during the growth phase of shrimp molt cycle. PMID- 21625496 TI - Molecular design, functional characterization and structural basis of a protein inhibitor against the HIV-1 pathogenicity factor Nef. AB - Increased spread of HIV-1 and rapid emergence of drug resistance warrants development of novel antiviral strategies. Nef, a critical viral pathogenicity factor that interacts with host cell factors but lacks enzymatic activity, is not targeted by current antiviral measures. Here we inhibit Nef function by simultaneously blocking several highly conserved protein interaction surfaces. This strategy, referred to as "wrapping Nef", is based on structure-function analyses that led to the identification of four target sites: (i) SH3 domain interaction, (ii) interference with protein transport processes, (iii) CD4 binding and (iv) targeting to lipid membranes. Screening combinations of Nef interacting domains, we developed a series of small Nef interacting proteins (NIs) composed of an SH3 domain optimized for binding to Nef, fused to a sequence motif of the CD4 cytoplasmic tail and combined with a prenylation signal for membrane association. NIs bind to Nef in the low nM affinity range, associate with Nef in human cells and specifically interfere with key biological activities of Nef. Structure determination of the Nef-inhibitor complex reveals the molecular basis for binding specificity. These results establish Nef-NI interfaces as promising leads for the development of potent Nef inhibitors. PMID- 21625497 TI - Novel two-component systems implied in antibiotic production in Streptomyces coelicolor. AB - The abundance of two-component systems (TCSs) in Streptomyces coelicolor A3(2) genome indicates their importance in the physiology of this soil bacteria. Currently, several TCSs have been related to antibiotic regulation, and the purpose in this study was the characterization of five TCSs, selected by sequence homology with the well-known absA1A2 system, that could also be associated with this important process. Null mutants of the five TCSs were obtained and two mutants (DeltaSCO1744/1745 and DeltaSCO4596/4597/4598) showed significant differences in both antibiotic production and morphological differentiation, and have been renamed as abr (antibiotic regulator). No detectable changes in antibiotic production were found in the mutants in the systems that include the ORFs SCO3638/3639, SCO3640/3641 and SCO2165/2166 in any of the culture conditions assayed. The system SCO1744/1745 (AbrA1/A2) was involved in negative regulation of antibiotic production, and acted also as a negative regulator of the morphological differentiation. By contrast, the system SCO4596/4597/4598 (AbrC1/C2/C3), composed of two histidine kinases and one response regulator, had positive effects on both morphological development and antibiotic production. Microarray analyses of the DeltaabrC1/C2/C3 and wild-type transcriptomes revealed downregulation of actII-ORF4 and cdaR genes, the actinorhodin and calcium dependent antibiotic pathway-specific regulators respectively. These results demonstrated the involvement of these new two-component systems in antibiotic production and morphological differentiation by different approaches. One is a pleiotropic negative regulator: abrA1/A2. The other one is a positive regulator composed of three elements, two histidine kinases and one response regulator: abrC1/C2/C3. PMID- 21625498 TI - Differential expression of CD163 on monocyte subsets in healthy and HIV-1 infected individuals. AB - CD163, a haptoglobin-hemoglobin (Hp-Hb) scavenger receptor, expressed by monocytes and macrophages, is important in resolution of inflammation. Age related non-AIDS co-morbidities in HIV-infected individuals, particularly dementia and cardiovascular disease, result in part from effects of HIV-1 infection on monocyte and macrophage biology. CD163 co-expression on CD14+CD16++ monocytes has been proposed as a useful biomarker for HIV-1 disease progression and the presence of HIV associated dementia. Here we investigated CD163 expression on monocyte subsets ex vivo, on cultured macrophages, and soluble in plasma, in the setting of HIV-1 infection. Whole blood immunophenotyping revealed CD163 expression on CD14++CD16- monocytes but not on CD14+CD16++ monocytes (P = 0.004), supported by CD163 mRNA levels. Incubation with M-CSF induced CD163 protein expression on CD14+CD16++ monocytes to the same extent as CD14++CD16- monocytes. CD163 expression on CD14++CD16+ monocytes from HIV-infected subjects was significantly higher than from uninfected individuals, with a trend towards increased expression on CD14++CD16- monocytes (P = 0.019 and 0.069 respectively), which is accounted for by HIV-1 therapy including protease inhibitors. Shedding of CD163 was shown to predominantly occur from the CD14++CD16- subset after Ficoll isolation and LPS stimulation. Soluble CD163 concentration in plasma from HIV-1 infected donors was similar to HIV-1 uninfected donors. Monocyte CD163 expression in HIV-1 infected patients showed a complicated relationship with classical measures of disease progression. Our findings clarify technical issues regarding CD163 expression on monocyte subsets and further elucidates its role in HIV-associated inflammation by demonstrating that CD163 is readily lost from CD14++CD16- monocytes and induced in pro-inflammatory CD14+CD16++ monocytes by M CSF. Our data show that all monocyte subsets are potentially capable of differentiating into CD163-expressing anti-inflammatory macrophages given appropriate stimuli. Levels of CD163 expression on monocytes may be a potential biomarker reflecting efforts by the immune system to resolve immune activation and inflammation in HIV-infected individuals. PMID- 21625499 TI - Real-time PCR improves Helicobacter pylori detection in patients with peptic ulcer bleeding. AB - BACKGROUND AND AIMS: Histological and rapid urease tests to detect H. pylori in biopsy specimens obtained during peptic ulcer bleeding episodes (PUB) often produce false-negative results. We aimed to examine whether immunohistochemistry and real-time PCR can improve the sensitivity of these biopsies. PATIENTS AND METHODS: We selected 52 histology-negative formalin-fixed paraffin-embedded biopsy specimens obtained during PUB episodes. Additional tests showed 10 were true negatives and 42 were false negatives. We also selected 17 histology positive biopsy specimens obtained during PUB to use as controls. We performed immunohistochemistry staining and real-time PCR for 16S rRNA, ureA, and 23S rRNA for H. pylori genes on all specimens. RESULTS: All controls were positive for H. pylori on all PCR assays and immunohistochemical staining. Regarding the 52 initially negative biopsies, all PCR tests were significantly more sensitive than immunohistochemical staining (p<0.01). Sensitivity and specificity were 55% and 80% for 16S rRNA PCR, 43% and 90% for ureA PCR, 41% and 80% for 23S rRNA PCR, and 7% and 100% for immunohistochemical staining, respectively. Combined analysis of PCR assays for two genes were significantly more sensitive than ureA or 23S rRNA PCR tests alone (p<0.05) and marginally better than 16S rRNA PCR alone. The best combination was 16S rRNA+ureA, with a sensitivity of 64% and a specificity of 80%. CONCLUSIONS: Real-time PCR improves the detection of H. pylori infection in histology-negative formalin-fixed paraffin-embedded biopsy samples obtained during PUB episodes. The low reported prevalence of H. pylori in PUB may be due to the failure of conventional tests to detect infection. PMID- 21625500 TI - Epigenetic alterations are critical for fear memory consolidation and synaptic plasticity in the lateral amygdala. AB - Epigenetic mechanisms, including histone acetylation and DNA methylation, have been widely implicated in hippocampal-dependent learning paradigms. Here, we have examined the role of epigenetic alterations in amygdala-dependent auditory Pavlovian fear conditioning and associated synaptic plasticity in the lateral nucleus of the amygdala (LA) in the rat. Using Western blotting, we first show that auditory fear conditioning is associated with an increase in histone H3 acetylation and DNMT3A expression in the LA, and that training-related alterations in histone acetylation and DNMT3A expression in the LA are downstream of ERK/MAPK signaling. Next, we show that intra-LA infusion of the histone deacetylase (HDAC) inhibitor TSA increases H3 acetylation and enhances fear memory consolidation; that is, long-term memory (LTM) is enhanced, while short term memory (STM) is unaffected. Conversely, intra-LA infusion of the DNA methyltransferase (DNMT) inhibitor 5-AZA impairs fear memory consolidation. Further, intra-LA infusion of 5-AZA was observed to impair training-related increases in H3 acetylation, and pre-treatment with TSA was observed to rescue the memory consolidation deficit induced by 5-AZA. In our final series of experiments, we show that bath application of either 5-AZA or TSA to amygdala slices results in significant impairment or enhancement, respectively, of long term potentiation (LTP) at both thalamic and cortical inputs to the LA. Further, the deficit in LTP following treatment with 5-AZA was observed to be rescued at both inputs by co-application of TSA. Collectively, these findings provide strong support that histone acetylation and DNA methylation work in concert to regulate memory consolidation of auditory fear conditioning and associated synaptic plasticity in the LA. PMID- 21625501 TI - Major-effect alleles at relatively few loci underlie distinct vernalization and flowering variation in Arabidopsis accessions. AB - We have explored the genetic basis of variation in vernalization requirement and response in Arabidopsis accessions, selected on the basis of their phenotypic distinctiveness. Phenotyping of F2 populations in different environments, plus fine mapping, indicated possible causative genes. Our data support the identification of FRI and FLC as candidates for the major-effect QTL underlying variation in vernalization response, and identify a weak FLC allele, caused by a Mutator-like transposon, contributing to flowering time variation in two N. American accessions. They also reveal a number of additional QTL that contribute to flowering time variation after saturating vernalization. One of these was the result of expression variation at the FT locus. Overall, our data suggest that distinct phenotypic variation in the vernalization and flowering response of Arabidopsis accessions is accounted for by variation that has arisen independently at relatively few major-effect loci. PMID- 21625502 TI - Visual personal familiarity in amnestic mild cognitive impairment. AB - BACKGROUND: Patients with amnestic mild cognitive impairment are at high risk for developing Alzheimer's disease. Besides episodic memory dysfunction they show deficits in accessing contextual knowledge that further specifies a general concept or helps to identify an object or a person. METHODOLOGY/PRINCIPAL FINDINGS: Using functional magnetic resonance imaging, we investigated the neural networks associated with the perception of personal familiar faces and places in patients with amnestic mild cognitive impairment and healthy control subjects. Irrespective of stimulus type, patients compared to control subjects showed lower activity in right prefrontal brain regions when perceiving personally familiar versus unfamiliar faces and places. Both groups did not show different neural activity when perceiving faces or places irrespective of familiarity. CONCLUSIONS/SIGNIFICANCE: Our data highlight changes in a frontal cortical network associated with knowledge-based personal familiarity among patients with amnestic mild cognitive impairment. These changes could contribute to deficits in social cognition and may reduce the patients' ability to transition from basic to complex situations and tasks. PMID- 21625503 TI - Hierarchical plant responses and diversity loss after nitrogen addition: testing three functionally-based hypotheses in the Inner Mongolia grassland. AB - BACKGROUND: Numerous studies have shown that nitrogen (N) deposition decreases biodiversity in terrestrial ecosystems. To explain the N-induced species loss, three functionally based hypotheses have been proposed: the aboveground competition hypothesis, the belowground competition hypothesis, and the total competition hypothesis. However, none of them is supported sufficiently by field experiments. A main challenge to testing these hypotheses is to ascertain the role of shoot and root competition in controlling plant responses to N enrichment. Simultaneously examining both aboveground and belowground responses in natural ecosystems is logistically complex, and has rarely been done. METHODOLOGY/PRINCIPAL FINDINGS: In a two-year N addition experiment conducted in a natural grassland ecosystem, we investigated both above- and belowground responses of plants at the individual, species, and community levels. Plants differed significantly in their responses to N addition across the different organizational levels. The community-level species loss was mainly due to the loss of perennial grasses and forbs, while the relative abundance of plant species was dependent mainly on individual-level responses. Plasticity in biomass allocation was much smaller within a species than between species, providing a biological basis for explaining the functionally based species loss. All species increased biomass allocation to aboveground parts, but species with high belowground allocations were replaced by those with high aboveground allocations, indicating that the increased aboveground competition was the key process responsible for the observed diversity loss after N addition in this grassland ecosystem. CONCLUSIONS/SIGNIFICANCE: Our findings shed new light on the validity of the three competing hypotheses concerning species loss in response to N enrichment. They also have important implications for predicting the future impacts of N deposition on the structure and functioning of terrestrial ecosystems. In addition, we have developed a new technique for ascertaining the roles of aboveground and belowground competition in determining plant responses to N fertilization. PMID- 21625504 TI - Lack of the delta subunit of RNA polymerase increases virulence related traits of Streptococcus mutans. AB - The delta subunit of the RNA polymerase, RpoE, maintains the transcriptional specificity in gram-positive bacteria. Lack of RpoE results in massive changes in the transcriptome of the human dental caries pathogen Streptococcus mutans. In this study, we analyzed traits of the DeltarpoE mutant which are important for biofilm formation and interaction with oral microorganisms and human cells and performed a global phenotypic analysis of its physiological functions. The DeltarpoE mutant showed higher self-aggregation compared to the wild type and coaggregated with other oral bacteria and Candida albicans. It formed a biofilm with a different matrix structure and an altered surface attachment. The amount of the cell surface antigens I/II SpaP and the glucosyltransferase GtfB was reduced. The DeltarpoE mutant displayed significantly stronger adhesion to human extracellular matrix components, especially to fibronectin, than the wild type. Its adhesion to human epithelial cells HEp-2 was reduced, probably due to the highly aggregated cell mass. The analysis of 1248 physiological traits using phenotype microarrays showed that the DeltarpoE mutant metabolized a wider spectrum of carbon sources than the wild type and had acquired resistance to antibiotics and inhibitory compounds with various modes of action. The reduced antigenicity, increased aggregation, adherence to fibronection, broader substrate spectrum and increased resistance to antibiotics of the DeltarpoE mutant reveal the physiological potential of S. mutans and show that some of its virulence related traits are increased. PMID- 21625505 TI - Influence of a brief episode of anesthesia during the induction of experimental brain trauma on secondary brain damage and inflammation. AB - It is unclear whether a single, brief, 15-minute episode of background anesthesia already modulates delayed secondary processes after experimental brain injury. Therefore, this study was designed to characterize three anesthesia protocols for their effect on molecular and histological study endpoints. Mice were randomly separated into groups that received sevoflurane (sevo), isoflurane (iso) or an intraperitoneal anesthetic combination (midazolam, fentanyl and medetomidine; comb) prior to traumatic brain injury (controlled cortical impact, CCI; 8 m/s, 1 mm impact depth, 3 mm diameter). Twenty-four hours after insult, histological brain damage, neurological function (via neurological severity score), cerebral inflammation (via real-time RT-PCR for IL6, COX-2, iNOS) and microglia (via immunohistochemical staining for Iba1) were determined. Fifteen minutes after CCI, the brain contusion volume did not differ between the anesthetic regimens (sevo = 17.9+/-5.5 mm(3); iso = 20.5+/-3.7 mm(3); comb = 19.5+/-4.6 mm(3)). Within 24 hours after injury, lesion size increased in all groups (sevo = 45.3+/ 9.0 mm(3); iso = 31.5+/-4.0 mm(3); comb = 44.2+/-6.2 mm(3)). Sevo and comb anesthesia resulted in a significantly larger contusion compared to iso, which was in line with the significantly better neurological function with iso (sevo = 4.6+/-1.3 pts.; iso = 3.9+/-0.8 pts.; comb = 5.1+/-1.6 pts.). The expression of inflammatory marker genes was not significantly different at 15 minutes and 24 hours after CCI. In contrast, significantly more Iba1-positive cells were present in the pericontusional region after sevo compared to comb anesthesia (sevo = 181+/-48/mm(3); iso = 150+/-36/mm(3); comb = 113+/-40/mm(3)). A brief episode of anesthesia, which is sufficient for surgical preparations of mice for procedures such as delivering traumatic brain injury, already has a significant impact on the extent of secondary brain damage. PMID- 21625506 TI - The C-terminal domain of the novel essential protein Gcp is critical for interaction with another essential protein YeaZ of Staphylococcus aureus. AB - Previous studies have demonstrated that the novel protein Gcp is essential for the viability of various bacterial species including Staphylococcus aureus; however, the reason why it is required for bacterial growth remains unclear. In order to explore the potential mechanisms of this essentiality, we performed RT PCR analysis and revealed that the gcp gene (sa1854) was co-transcribed with sa1855, yeaZ (sa1856) and sa1857 genes, indicating these genes are located in the same operon. Furthermore, we demonstrated that Gcp interacts with YeaZ using a yeast two-hybrid (Y2H) system and in vitro pull down assays. To characterize the Gcp-YeaZ interaction, we performed alanine scanning mutagenesis on the residues of C-terminal segment of Gcp. We found that the mutations of the C-terminal Y317 F322 region abolished the interaction of Gcp and YeaZ, and the mutations of the D324-N329 and S332-Y336 regions alleviated Gcp binding to YeaZ. More importantly, we demonstrated that these key regions of Gcp are also necessary for the bacterial survival since these mutated Gcp could not complement the depletion of endogenous Gcp. Taken together, our data suggest that the interaction of Gcp and YeaZ may contribute to the essentiality of Gcp for S. aureus survival. Our findings provide new insights into the potential mechanisms and biological functions of this novel essential protein. PMID- 21625507 TI - Distinctive responsiveness to stromal signaling accompanies histologic grade programming of cancer cells. AB - Whether stromal components facilitate growth, invasion, and dissemination of cancer cells or suppress neoplastic lesions from further malignant progression is a continuing conundrum in tumor biology. Conceptualizing a dynamic picture of tumorigenesis is complicated by inter-individual heterogeneity. In the post genomic era, unraveling such complexity remains a challenge for the cancer biologist. Towards establishing a functional association between cellular crosstalk and differential cancer aggressiveness, we identified a signature of malignant breast epithelial response to stromal signaling. Proximity to fibroblasts resulted in gene transcript alterations of >2-fold for 107 probes, collectively designated as Fibroblast Triggered Gene Expression in Tumor (FTExT). The hazard ratio predicted by the FTExT classifier for distant relapse in patients with intermediate and high grade breast tumors was significant compared to routine clinical variables (dataset 1, n = 258, HR--2.11, 95% CI 1.17-3.80, p value 0.01; dataset 2, n = 171, HR--3.07, 95% CI 1.21-7.83, p-value 0.01). Biofunctions represented by FTExT included inflammatory signaling, free radical scavenging, cell death, and cell proliferation. Unlike genes of the 'proliferation cluster', which are overexpressed in aggressive primary tumors, FTExT genes were uniquely repressed in such cases. As proof of concept for our correlative findings, which link stromal-epithelial crosstalk and tumor behavior, we show a distinctive differential in stromal impact on prognosis-defining functional endpoints of cell cycle progression, and resistance to therapy-induced growth arrest and apoptosis in low vs. high grade cancer cells. Our experimental data thus reveal aspects of 'paracrine cooperativity' that are exclusively contingent upon the histopathologically defined grade of interacting tumor epithelium, and demonstrate that epithelial responsiveness to the tumor microenvironment is a deterministic factor underlying clinical outcome. In this light, early attenuation of epithelial-stromal crosstalk could improve the management of cases prone to be clinically challenging. PMID- 21625508 TI - MoSfl1 is important for virulence and heat tolerance in Magnaporthe oryzae. AB - The formation of appressoria, specialized plant penetration structures of Magnaporthe oryzae, is regulated by the MST11-MST7-PMK1 MAP kinase cascade. One of its downstream transcription factor, MST12, is important for penetration and invasive growth but dispensable for appressorium formation. To identify additional downstream targets that are regulated by Pmk1, in this study we performed phosphorylation assays with a protein microarray composed of 573 M. oryzae transcription factor (TF) genes. Three of the TF genes phosphorylated by Pmk1 in vitro were further analyzed by coimmunoprecipitation assays. One of them, MoSFL1, was found to interact with Pmk1 in vivo. Like other Sfl1 orthologs, the MoSfl1 protein has the HSF-like domain. When expressed in yeast, MoSFL1 functionally complemented the flocculation defects of the sfl1 mutant. In M. oryzae, deletion of MoSFl1 resulted in a significant reduction in virulence on rice and barley seedlings. Consistent with this observation, the Mosfl1 mutant was defective in invasive growth in penetration assays with rice leaf sheaths. In comparison with that of vegetative hyphae, the expression level of MoSFL1 was increased in appressoria and infected rice leaves. The Mosfl1 mutant also had increased sensitivity to elevated temperatures. In CM cultures of the Mosfl1 and pmk1 mutants grown at 30 degrees C, the production of aerial hyphae and melanization were reduced but their growth rate was not altered. When assayed by qRT-PCR, the transcription levels of the MoHSP30 and MoHSP98 genes were reduced 10- and 3-fold, respectively, in the Mosfl1 mutant. SFL1 orthologs are conserved in filamentous ascomycetes but none of them have been functionally characterized in non-Saccharomycetales fungi. MoSfl1 has one putative MAPK docking site and three putative MAPK phosphorylation sites. Therefore, it may be functionally related to Pmk1 in the regulation of invasive growth and stress responses in M. oryzae. PMID- 21625509 TI - Presentation and outcome of tuberculous meningitis in a high HIV prevalence setting. AB - BACKGROUND: Mycobacterium tuberculosis is a common, devastating cause of meningitis in HIV-infected persons. Due to international rollout programs, access to antiretroviral therapy (ART) is increasing globally. Starting patients with HIV-associated tuberculous meningitis (TBM) on ART during tuberculosis (TB) treatment may increase survival in these patients. We undertook this study to describe causes of meningitis at a secondary-level hospital in a high HIV/TB co infection setting and to determine predictors of mortality in patients with TBM. METHODS: A retrospective review of cerebrospinal fluid findings and clinical records over a six-month period (March 2009-August 2009). Definite, probable and possible TBM were diagnosed according to published case definitions. RESULTS: TBM was diagnosed in 120/211 patients (57%) with meningitis. In 106 HIV-infected patients with TBM, six-month all-cause mortality was lower in those who received antiretroviral therapy (ART) during TB treatment; hazard ratio = 0.30 (95% CI = 0.08-0.82). Factors associated with inpatient mortality in HIV-infected patients were 1) low CD4(+) count at presentation; adjusted odds ratio (AOR) = 1.4 (95% confidence interval [CI] = 1.03-1.96) per 50 cells/uL drop in CD4(+) count and, 2) higher British Medical Research Council TBM disease grade (2 or 3 versus 1); AOR = 4.8 (95% CI = 1.45-15.87). INTERPRETATION: Starting ART prior to or during TB treatment may be associated with lower mortality in patients with HIV associated TBM. Advanced HIV and worse stage of TBM disease predict in-hospital mortality in patients presenting with TBM. PMID- 21625510 TI - Secretor genotype (FUT2 gene) is strongly associated with the composition of Bifidobacteria in the human intestine. AB - Intestinal microbiota plays an important role in human health, and its composition is determined by several factors, such as diet and host genotype. However, thus far it has remained unknown which host genes are determinants for the microbiota composition. We studied the diversity and abundance of dominant bacteria and bifidobacteria from the faecal samples of 71 healthy individuals. In this cohort, 14 were non-secretor individuals and the remainders were secretors. The secretor status is defined by the expression of the ABH and Lewis histo-blood group antigens in the intestinal mucus and other secretions. It is determined by fucosyltransferase 2 enzyme, encoded by the FUT2 gene. Non-functional enzyme resulting from a nonsense mutation in the FUT2 gene leads to the non-secretor phenotype. PCR-DGGE and qPCR methods were applied for the intestinal microbiota analysis. Principal component analysis of bifidobacterial DGGE profiles showed that the samples of non-secretor individuals formed a separate cluster within the secretor samples. Moreover, bifidobacterial diversity (p<0.0001), richness (p<0.0003), and abundance (p<0.05) were significantly reduced in the samples from the non-secretor individuals as compared with those from the secretor individuals. The non-secretor individuals lacked, or were rarely colonized by, several genotypes related to B. bifidum, B. adolescentis and B. catenulatum/pseudocatenulatum. In contrast to bifidobacteria, several bacterial genotypes were more common and the richness (p<0.04) of dominant bacteria as detected by PCR-DGGE was higher in the non-secretor individuals than in the secretor individuals. We showed that the diversity and composition of the human bifidobacterial population is strongly associated with the histo-blood group ABH secretor/non-secretor status, which consequently appears to be one of the host genetic determinants for the composition of the intestinal microbiota. This association can be explained by the difference between the secretor and non secretor individuals in their expression of ABH and Lewis glycan epitopes in the mucosa. PMID- 21625511 TI - Clinical Clostridium difficile: clonality and pathogenicity locus diversity. AB - Clostridium difficile infection (CDI) is an important cause of mortality and morbidity in healthcare settings. The major virulence determinants are large clostridial toxins, toxin A (tcdA) and toxin B (tcdB), encoded within the pathogenicity locus (PaLoc). Isolates vary in pathogenicity from hypervirulent PCR-ribotypes 027 and 078 with high mortality, to benign non-toxigenic strains carried asymptomatically. The relative pathogenicity of most toxigenic genotypes is still unclear, but may be influenced by PaLoc genetic variant. This is the largest study of C. difficile molecular epidemiology performed to date, in which a representative collection of recent isolates (n = 1290) from patients with CDI in Oxfordshire, UK, was genotyped by multilocus sequence typing. The population structure was described using NeighborNet and ClonalFrame. Sequence variation within toxin B (tcdB) and its negative regulator (tcdC), was mapped onto the population structure. The 69 Sequence Types (ST) showed evidence for homologous recombination with an effect on genetic diversification four times lower than mutation. Five previously recognised genetic groups or clades persisted, designated 1 to 5, each having a strikingly congruent association with tcdB and tcdC variants. Hypervirulent ST-11 (078) was the only member of clade 5, which was divergent from the other four clades within the MLST loci. However, it was closely related to the other clades within the tcdB and tcdC loci. ST-11 (078) may represent a divergent formerly non-toxigenic strain that acquired the PaLoc (at least) by genetic recombination. This study focused on human clinical isolates collected from a single geographic location, to achieve a uniquely high density of sampling. It sets a baseline of MLST data for future comparative studies investigating genotype virulence potential (using clinical severity data for these isolates), possible reservoirs of human CDI, and the evolutionary origins of hypervirulent strains. PMID- 21625512 TI - Expression of a malarial Hsp70 improves defects in chaperone-dependent activities in ssa1 mutant yeast. AB - Plasmodium falciparum causes the most virulent form of malaria and encodes a large number of molecular chaperones. Because the parasite encounters radically different environments during its lifecycle, many members of this chaperone ensemble may be essential for P. falciparum survival. Therefore, Plasmodium chaperones represent novel therapeutic targets, but to establish the mechanism of action of any developed therapeutics, it is critical to ascertain the functions of these chaperones. To this end, we report the development of a yeast expression system for PfHsp70-1, a P. falciparum cytoplasmic chaperone. We found that PfHsp70-1 repairs mutant growth phenotypes in yeast strains lacking the two primary cytosolic Hsp70s, SSA1 and SSA2, and in strains harboring a temperature sensitive SSA1 allele. PfHsp70-1 also supported chaperone-dependent processes such as protein translocation and ER associated degradation, and ameliorated the toxic effects of oxidative stress. By introducing engineered forms of PfHsp70-1 into the mutant strains, we discovered that rescue requires PfHsp70-1 ATPase activity. Together, we conclude that yeast can be co-opted to rapidly uncover specific cellular activities mediated by malarial chaperones. PMID- 21625513 TI - Sour taste responses in mice lacking PKD channels. AB - BACKGROUND: The polycystic kidney disease-like ion channel PKD2L1 and its associated partner PKD1L3 are potential candidates for sour taste receptors. PKD2L1 is expressed in type III taste cells that respond to sour stimuli and genetic elimination of cells expressing PKD2L1 substantially reduces chorda tympani nerve responses to sour taste stimuli. However, the contribution of PKD2L1 and PKD1L3 to sour taste responses remains unclear. METHODOLOGY/PRINCIPAL FINDINGS: We made mice lacking PKD2L1 and/or PKD1L3 gene and investigated whole nerve responses to taste stimuli in the chorda tympani or the glossopharyngeal nerve and taste responses in type III taste cells. In mice lacking PKD2L1 gene, chorda tympani nerve responses to sour, but not sweet, salty, bitter, and umami tastants were reduced by 25-45% compared with those in wild type mice. In contrast, chorda tympani nerve responses in PKD1L3 knock-out mice and glossopharyngeal nerve responses in single- and double-knock-out mice were similar to those in wild type mice. Sour taste responses of type III fungiform taste cells (GAD67-expressing taste cells) were also reduced by 25-45% by elimination of PKD2L1. CONCLUSIONS/SIGNIFICANCE: These findings suggest that PKD2L1 partly contributes to sour taste responses in mice and that receptors other than PKDs would be involved in sour detection. PMID- 21625514 TI - An extensive network of information flow through the B1b/c intersubunit bridge of the yeast ribosome. AB - Yeast ribosomal proteins L11 and S18 form a dynamic intersubunit interaction called the B1b/c bridge. Recent high resolution images of the ribosome have enabled targeting of specific residues in this bridge to address how distantly separated regions within the large and small subunits of the ribosome communicate with each other. Mutations were generated in the L11 side of the B1b/c bridge with a particular focus on disrupting the opposing charge motifs that have previously been proposed to be involved in subunit ratcheting. Mutants had wide ranging effects on cellular viability and translational fidelity, with the most pronounced phenotypes corresponding to amino acid changes resulting in alterations of local charge properties. Chemical protection studies of selected mutants revealed rRNA structural changes in both the large and small subunits. In the large subunit rRNA, structural changes mapped to Helices 39, 80, 82, 83, 84, and the peptidyltransferase center. In the small subunit rRNA, structural changes were identified in helices 30 and 42, located between S18 and the decoding center. The rRNA structural changes correlated with charge-specific alterations to the L11 side of the B1b/c bridge. These analyses underscore the importance of the opposing charge mechanism in mediating B1b/c bridge interactions and suggest an extensive network of information exchange between distinct regions of the large and small subunits. PMID- 21625515 TI - PrP(Sc)-specific antibodies with the ability to immunodetect prion oligomers. AB - The development of antibodies with binding capacity towards soluble oligomeric forms of PrPSc recognised in the aggregation process in early stage of the disease would be of paramount importance in diagnosing prion diseases before extensive neuropathology has ensued. As blood transfusion appears to be efficient in the transmission of the infectious prion agent, there is an urgent need to develop reagents that would specifically recognize oligomeric forms of the abnormally folded prion protein, PrPSc.To that end, we show that anti-PrP monoclonal antibodies (called PRIOC mAbs) derived from mice immunised with native PrP-coated microbeads are able to immunodetect oligomers/multimers of PrPSc. Oligomer-specific immunoreactivity displayed by these PRIOC mAbs was demonstrated as large aggregates of immunoreactive deposits in prion-permissive neuroblastoma cell lines but not in equivalent non-infected or prn-p(0/0) cell lines. In contrast, an anti-monomer PrP antibody displayed diffuse immunoreactivity restricted to the cell membrane. Furthermore, our PRIOC mAbs did not display any binding with monomeric recombinant and cellular prion proteins but strongly detected PrPSc oligomers as shown by a newly developed sensitive and specific ELISA. Finally, PrioC antibodies were also able to bind soluble oligomers formed of Abeta and alpha-synuclein. These findings demonstrate the potential use of anti-prion antibodies that bind PrPSc oligomers, recognised in early stage of the disease, for the diagnosis of prion diseases in blood and other body fluids. PMID- 21625516 TI - Significance of thymosin beta4 and implication of PINCH-1-ILK-alpha-parvin (PIP) complex in human dilated cardiomyopathy. AB - Myocardial remodeling is a major contributor in the development of heart failure (HF) after myocardial infarction (MI). Integrin-linked kinase (ILK), LIM-only adaptor PINCH-1, and alpha-parvin are essential components of focal adhesions (FAs), which are highly expressed in the heart. ILK binds tightly to PINCH-1 and alpha-parvin, which regulates FA assembly and promotes cell survival via the activation of the kinase Akt. Mice lacking ILK, PINCH or alpha-parvin have been shown to develop severe defects in the heart, suggesting that these proteins play a critical role in heart function. Utilizing failing human heart tissues (dilated cardiomyopathy, DCM), we found a 2.27-fold (p<0.001) enhanced expression of PINCH, 4 fold for alpha-parvin, and 10.5 fold (p<0.001) for ILK as compared to non-failing (NF) counterparts. No significant enhancements were found for the PINCH isoform PINCH-2 and parvin isoform beta-parvin. Using a co immunoprecipitation method, we also found that the PINCH-1-ILK-alpha-parvin (PIP) complex and Akt activation were significantly up-regulated. These observations were further corroborated with the mouse myocardial infarction (MI) and transaortic constriction (TAC) model. Thymosin beta4 (Tbeta4), an effective cell penetrating peptide for treating MI, was found to further enhance the level of PIP components and Akt activation, while substantially suppressing NF-kappaB activation and collagen expression--the hallmarks of cardiac fibrosis. In the presence of an Akt inhibitor, wortmannin, we show that Tbeta4 had a decreased effect in protecting the heart from MI. These data suggest that the PIP complex and activation of Akt play critical roles in HF development. Tbeta4 treatment likely improves cardiac function by enhancing PIP mediated Akt activation and suppressing NF-kappaB activation and collagen-mediated fibrosis. These data provide significant insight into the role of the PIP-Akt pathway and its regulation by Tbeta4 treatment in post-MI. PMID- 21625517 TI - Interictal functional connectivity of human epileptic networks assessed by intracerebral EEG and BOLD signal fluctuations. AB - In this study, we aimed to demonstrate whether spontaneous fluctuations in the blood oxygen level dependent (BOLD) signal derived from resting state functional magnetic resonance imaging (fMRI) reflect spontaneous neuronal activity in pathological brain regions as well as in regions spared by epileptiform discharges. This is a crucial issue as coherent fluctuations of fMRI signals between remote brain areas are now widely used to define functional connectivity in physiology and in pathophysiology. We quantified functional connectivity using non-linear measures of cross-correlation between signals obtained from intracerebral EEG (iEEG) and resting-state functional MRI (fMRI) in 5 patients suffering from intractable temporal lobe epilepsy (TLE). Functional connectivity was quantified with both modalities in areas exhibiting different electrophysiological states (epileptic and non affected regions) during the interictal period. Functional connectivity as measured from the iEEG signal was higher in regions affected by electrical epileptiform abnormalities relative to non-affected areas, whereas an opposite pattern was found for functional connectivity measured from the BOLD signal. Significant negative correlations were found between the functional connectivities of iEEG and BOLD signal when considering all pairs of signals (theta, alpha, beta and broadband) and when considering pairs of signals in regions spared by epileptiform discharges (in broadband signal). This suggests differential effects of epileptic phenomena on electrophysiological and hemodynamic signals and/or an alteration of the neurovascular coupling secondary to pathological plasticity in TLE even in regions spared by epileptiform discharges. In addition, indices of directionality calculated from both modalities were consistent showing that the epileptogenic regions exert a significant influence onto the non epileptic areas during the interictal period. This study shows that functional connectivity measured by iEEG and BOLD signals give complementary but sometimes inconsistent information in TLE. PMID- 21625518 TI - Identification of metabolites in the normal ovary and their transformation in primary and metastatic ovarian cancer. AB - In this study, we characterized the metabolome of the human ovary and identified metabolic alternations that coincide with primary epithelial ovarian cancer (EOC) and metastatic tumors resulting from primary ovarian cancer (MOC) using three analytical platforms: gas chromatography mass spectrometry (GC/MS) and liquid chromatography tandem mass spectrometry (LC/MS/MS) using buffer systems and instrument settings to catalog positive or negative ions. The human ovarian metabolome was found to contain 364 biochemicals and upon transformation of the ovary caused changes in energy utilization, altering metabolites associated with glycolysis and beta-oxidation of fatty acids--such as carnitine (1.79 fold in EOC, p<0.001; 1.88 fold in MOC, p<0.001), acetylcarnitine (1.75 fold in EOC, p<0.001; 2.39 fold in MOC, p<0.001), and butyrylcarnitine (3.62 fold, p<0.0094 in EOC; 7.88 fold, p<0.001 in MOC). There were also significant changes in phenylalanine catabolism marked by increases in phenylpyruvate (4.21 fold; p = 0.0098) and phenyllactate (195.45 fold; p<0.0023) in EOC. Ovarian cancer also displayed an enhanced oxidative stress response as indicated by increases in 2 aminobutyrate in EOC (1.46 fold, p = 0.0316) and in MOC (2.25 fold, p<0.001) and several isoforms of tocopherols. We have also identified novel metabolites in the ovary, specifically N-acetylasparate and N-acetyl-aspartyl-glutamate, whose role in ovarian physiology has yet to be determined. These data enhance our understanding of the diverse biochemistry of the human ovary and demonstrate metabolic alterations upon transformation. Furthermore, metabolites with significant changes between groups provide insight into biochemical consequences of transformation and are candidate biomarkers of ovarian oncogenesis. Validation studies are warranted to determine whether these compounds have clinical utility in the diagnosis or clinical management of ovarian cancer patients. PMID- 21625519 TI - The Salmonella enterica PhoP directly activates the horizontally acquired SPI-2 gene sseL and is functionally different from a S. bongori ortholog. AB - To establish a successful infection within the host, a pathogen must closely regulate multiple virulence traits to ensure their accurate temporal and spatial expression. As a highly adapted intracellular pathogen, Salmonella enterica has acquired during its evolution various virulence genes via numerous lateral transfer events, including the acquisition of the Salmonella Pathogenicity Island 2 (SPI-2) and its associated effectors. Beneficial use of horizontally acquired genes requires that their expression is effectively coordinated with the already existing virulence programs and the regulatory set-up in the bacterium. As an example for such a mechanism, we show here that the ancestral PhoPQ system of Salmonella enterica is able to regulate directly the SPI-2 effector gene sseL (encoding a secreted deubiquitinase) in an SsrB-independent manner and that PhoP plays a part in a feed-forward regulatory loop, which fine-tunes the cellular level of SseL. Additionally, we demonstrate the presence of conserved cis regulatory elements in the promoter region of sseL and show direct binding of purified PhoP to this region. Interestingly, in contrast to the S. enterica PhoP, an ortholog regulator from a S. bongori SARC 12 strain was found to be impaired in promoting transcription of sseL and other genes from the PhoP regulon. These findings have led to the identification of a previously uncharacterized residue in the DNA-binding domain of PhoP, which is required for the transcriptional activation of PhoP regulated genes in Salmonella spp. Collectively our data demonstrate an interesting interface between the acquired SsrB regulon and the ancestral PhoPQ regulatory circuit, provide novel insights into the function of PhoP, and highlight a mechanism of regulatory integration of horizontally acquired genes into the virulence network of Salmonella enterica. PMID- 21625520 TI - Isolation and characterization of porcine amniotic fluid-derived multipotent stem cells. AB - The aim of this study was to isolate and characterize porcine amniotic fluid derived multipotent stem cells (pAF-MSC). The porcine amniotic fluid (AF) from the amniotic cavity of pregnant gilts in the early stages of gestation (at E35) was collected and centrifuged for 5-10 min at 400 g to pellet cells. The primary culture of AF showed the multiple cell types, including the epithelial-like cells and fibroblast-like cells. By culturing in AMM medium for 6 to 8 days, the epithelial-like cells disappeared and the remaining cells presented the fibroblastoid morphology. The doubling time of pAF-MSCs was about 34.6 h, and the cells had been continually cultured over 60 passages in vitro. The flow cytometry results showed that pAF-MSCs were positive for CD44, CD117 and CD166, but negative for CD34, CD45 and CD54. Meanwhile, pAF-MSCs expressed ES cell markers, such as Oct4, Nanog, SSEA4, Tra-1-60 and Tra-1-81. The ratio of CD117(+) CD44(+) cells accounted for 98% of pAF-MSCs population. Three germ layer markers, including FGF5 (ectodermal marker), AFP (endodermal marker) and Bra (mesodermal marker), were detected in embryoid bodies derived from pAF-MSCs. Under the different induction conditions, the pAF-MSCs were capable of differentiating into neurocytes, adipocytes and beating cardiomyocytes. Furthermore, the pAF-MSCs didn't form teratoma when injected into immunodeficiency mice. These optimal features of pAF-MSCs provide an excellent alternative stem cell resource for potential cell therapy in regenerative medicine and transgenic animals. PMID- 21625521 TI - Long-lasting inhibitory effects of fetal liver mesenchymal stem cells on T lymphocyte proliferation. AB - Human bone marrow mesenchymal stem cells (BM-MSC) are multipotent progenitor cells that have transient immunomodulatory properties on Natural Killer (NK) cells, Dendritic Cells (DC), and T cells. This study compared the use of MSC isolated from bone marrow and fetal liver (FL-MSC) to determine which displayed the most efficient immunosuppressive effects on T cell activation. Although both types of MSC exhibit similar phenotype profile, FL-MSC displays a much more extended in vitro life-span and immunomodulatory properties. When co-cultured with CD3/CD28-stimulated T cells, both BM-MSC and FL-MSC affected T cell proliferation by inhibiting their entry into the cell cycle, by inducing the down regulation of phospho-retinoblastoma (pRb), cyclins A and D1, as well as up regulating p27(kip1) expression. The T cell inhibition by MSC was not due to the soluble HLA-G5 isoform, but to the surface expression of HLA-G1, as shown by the need of cell-cell contact and by the use of neutralizing anti-HLA-G antibodies. To note, in a HLA-G-mediated fashion, MSC facilitated the expansion of a CD4(low)/CD8(low) T subset that had decreased secretion of IFN-gamma, and an induced secretion of the immunomodulatory cytokine IL-10. Because of their longer lasting in vitro immunosuppressive properties, mainly mediated by HLA-G, and their more efficient induction of IL-10 production and T cell apoptosis, fetal liver MSC could be considered a new tool for MSC therapy to prevent allograft rejection. PMID- 21625522 TI - Dynamics of the STAT3 transcription factor: nuclear import dependent on Ran and importin-beta1. AB - The signal transducer and activator of transcription-3 (STAT3) induces transcription of genes that control differentiation, inflammation, proliferation, and tumor cell invasion. Cytokines such as interleukin-6 and interferon stimulate the specific tyrosine phosphorylation of STAT3, which confers its ability to bind consensus DNA targets. In addition, unphosphorylated STAT3 has been demonstrated to induce specific gene expression. STAT3 must gain entrance to the nucleus to impact transcription, however access to the nucleus is a tightly regulated process. Because nuclear trafficking is critical to the function of STAT3, we investigated the molecular mechanisms by which STAT3 is imported to the nucleus. Live cell imaging techniques were used with STAT3 tagged with green fluorescence protein (GFP) or photoactivatable GFP to follow the cellular dynamics of both unphosphorylated and tyrosine phosphorylated forms. Cytokine activation did not alter the rate of STAT3 nuclear import or nuclear export. In addition, Forster resonance energy transfer experiments revealed homomeric interaction of unphosphorylated STAT3 dependent on its amino terminus, but this dimerization is not necessary for its nuclear import. Previous work demonstrated the adapter importin-alpha3 binds to STAT3 and is required for nuclear import. To determine whether STAT3 nuclear import is mediated by the importin-alpha/importin-beta1 heterodimer, the effects of siRNA to importin-beta1 were evaluated. Results indicate STAT3 nuclear import is dependent on the function of importin-beta1. Since the Ran GTPase is necessary to bind importin-beta1 in the nucleus for release of importin-alpha-cargo, the effect of a GTPase deficient mutant of Ran was tested. Expression of the Ran interfering mutant inhibited STAT3 nuclear import. This study defines importin-alpha/importin-beta1/Ran as the molecular mechanism by which STAT3 traffics to the nucleus. PMID- 21625523 TI - Expression of OATP family members in hormone-related cancers: potential markers of progression. AB - The organic anion transporting polypeptide (OATP) family of transporters has been implicated in prostate cancer disease progression probably by transporting hormones or drugs. In this study, we aimed to elucidate the expression, frequency, and relevance of OATPs as a biomarker in hormone-dependent cancers. We completed a study examining SLCO1B3, SLCO1B1 and SLCO2B1 mRNA expression in 381 primary, independent patient samples representing 21 cancers and normal tissues. From a separate cohort, protein expression of OATP1B3 was examined in prostate, colon, and bladder tissue. Based on expression frequency, SLCO2B1 was lower in liver cancer (P = 0.04) which also trended lower with decreasing differentiation (P = 0.004) and lower magnitude in pancreatic cancer (P = 0.05). SLCO2B1 also had a higher frequency in thyroid cancer (67%) than normal (0%) and expression increased with stage (P = 0.04). SLCO1B3 was expressed in 52% of cancerous prostate samples and increased SLCO1B3 expression trended with higher Gleason score (P = 0.03). SLCO1B3 expression was also higher in testicular cancer (P = 0.02). SLCO1B1 expression was lower in liver cancer (P = 0.04) which trended lower with liver cancer grade (P = 0.0004) and higher with colon cancer grade (P = 0.05). Protein expression of OATP1B3 was examined in normal and cancerous prostate, colon, and bladder tissue samples from an independent cohort. The results were similar to the transcription data, but showed distinct localization. OATPs correlate to differentiation in certain hormone-dependent cancers, thus may be useful as biomarkers for assessing clinical treatment and stage of disease. PMID- 21625524 TI - Chronic oral infection with Porphyromonas gingivalis accelerates atheroma formation by shifting the lipid profile. AB - BACKGROUND: Recent studies have suggested that periodontal disease increases the risk of atherothrombotic disease. Atherosclerosis has been characterized as a chronic inflammatory response to cholesterol deposition in the arteries. Although several studies have suggested that certain periodontopathic bacteria accelerate atherogenesis in apolipoprotein E-deficient mice, the mechanistic link between cholesterol accumulation and periodontal infection-induced inflammation is largely unknown. METHODOLOGY/PRINCIPAL FINDINGS: We orally infected C57BL/6 and C57BL/6.KOR-Apoe(shl) (B6.Apoeshl) mice with Porphyromonas gingivalis, which is a representative periodontopathic bacterium, and evaluated atherogenesis, gene expression in the aorta and liver and systemic inflammatory and lipid profiles in the blood. Furthermore, the effect of lipopolysaccharide (LPS) from P. gingivalis on cholesterol transport and the related gene expression was examined in peritoneal macrophages. Alveolar bone resorption and elevation of systemic inflammatory responses were induced in both strains. Despite early changes in the expression of key genes involved in cholesterol turnover, such as liver X receptor and ATP-binding cassette A1, serum lipid profiles did not change with short-term infection. Long-term infection was associated with a reduction in serum high-density lipoprotein (HDL) cholesterol but not with the development of atherosclerotic lesions in wild-type mice. In B6.Apoeshl mice, long-term infection resulted in the elevation of very low-density lipoprotein (VLDL), LDL and total cholesterols in addition to the reduction of HDL cholesterol. This shift in the lipid profile was concomitant with a significant increase in atherosclerotic lesions. Stimulation with P. gingivalis LPS induced the change of cholesterol transport via targeting the expression of LDL receptor-related genes and resulted in the disturbance of regulatory mechanisms of the cholesterol level in macrophages. CONCLUSIONS/SIGNIFICANCE: Periodontal infection itself does not cause atherosclerosis, but it accelerates it by inducing systemic inflammation and deteriorating lipid metabolism, particularly when underlying hyperlidemia or susceptibility to hyperlipidemia exists, and it may contribute to the development of coronary heart disease. PMID- 21625525 TI - IgE immune complexes stimulate an increase in lung mast cell progenitors in a mouse model of allergic airway inflammation. AB - Mast cell numbers and allergen specific IgE are increased in the lungs of patients with allergic asthma and this can be reproduced in mouse models. The increased number of mast cells is likely due to recruitment of mast cell progenitors that mature in situ. We hypothesized that formation of IgE immune complexes in the lungs of sensitized mice increase the migration of mast cell progenitors to this organ. To study this, a model of allergic airway inflammation where mice were immunized with ovalbumin (OVA) in alum twice followed by three daily intranasal challenges of either OVA coupled to trinitrophenyl (TNP) alone or as immune complexes with IgE-anti-TNP, was used. Mast cell progenitors were quantified by a limiting dilution assay. IgE immune complex challenge of sensitized mice elicited three times more mast cell progenitors per lung than challenge with the same dose of antigen alone. This dose of antigen challenge alone did not increase the levels of mast cell progenitors compared to unchallenged mice. IgE immune complex challenge of sensitized mice also enhanced the frequency of mast cell progenitors per 10(6) mononuclear cells by 2.1-fold. The enhancement of lung mast cell progenitors by IgE immune complex challenge was lost in FcRgamma deficient mice but not in CD23 deficient mice. Our data show that IgE immune complex challenge enhances the number of mast cell progenitors in the lung through activation of an Fc receptor associated with the FcRgamma chain. This most likely takes place via activation of FcepsilonRI, although activation via FcgammaRIV or a combination of the two receptors cannot be excluded. IgE immune complex-mediated enhancement of lung MCp numbers is a new reason to target IgE in therapies against allergic asthma. PMID- 21625526 TI - Var2CSA minimal CSA binding region is located within the N-terminal region. AB - Var2CSA, a key molecule linked with pregnancy-associated malaria (PAM), causes sequestration of Plasmodium falciparum infected erythrocytes (PEs) in the placenta by adhesion to chondroitin sulfate A (CSA). Var2CSA possesses a 300 kDa extracellular region composed of six Duffy-binding like (DBL) domains and a cysteine-rich interdomain region (CIDRpam) module. Although initial studies implicated several individual var2CSA DBL domains as important for adhesion of PEs to CSA, new studies revealed that these individual domains lack both the affinity and specificity displayed by the full-length extracellular region. Indeed, recent evidence suggests the presence of a single CSA-binding site formed by a higher-order domain organization rather than several independent binding sites located on the different domains. Here, we search for the minimal binding region within var2CSA that maintains high affinity and specificity for CSA binding, a characteristic feature of the full-length extracellular region. Accordingly, truncated recombinant var2CSA proteins comprising different domain combinations were expressed and their binding characteristics assessed against different sulfated glycosaminoglycans (GAGs). Our results indicate that the smallest region within var2CSA with similar binding properties to those of the full-length var2CSA is DBL1X-3X. We also demonstrate that inhibitory antibodies raised in rabbit against the full-length DBL1X-6epsilon target principally DBL3X and, to a lesser extent, DBL5epsilon. Taken together, our results indicate that efforts should focus on the DBL1X-3X region for developing vaccine and therapeutic strategies aimed at combating PAM. PMID- 21625527 TI - Transition of Plasmodium sporozoites into liver stage-like forms is regulated by the RNA binding protein Pumilio. AB - Many eukaryotic developmental and cell fate decisions that are effected post transcriptionally involve RNA binding proteins as regulators of translation of key mRNAs. In malaria parasites (Plasmodium spp.), the development of round, non motile and replicating exo-erythrocytic liver stage forms from slender, motile and cell-cycle arrested sporozoites is believed to depend on environmental changes experienced during the transmission of the parasite from the mosquito vector to the vertebrate host. Here we identify a Plasmodium member of the RNA binding protein family PUF as a key regulator of this transformation. In the absence of Pumilio-2 (Puf2) sporozoites initiate EEF development inside mosquito salivary glands independently of the normal transmission-associated environmental cues. Puf2- sporozoites exhibit genome-wide transcriptional changes that result in loss of gliding motility, cell traversal ability and reduction in infectivity, and, moreover, trigger metamorphosis typical of early Plasmodium intra-hepatic development. These data demonstrate that Puf2 is a key player in regulating sporozoite developmental control, and imply that transformation of salivary gland resident sporozoites into liver stage-like parasites is regulated by a post transcriptional mechanism. PMID- 21625528 TI - Perioperative use of herbal, complementary, and over the counter medicines in plastic surgery patients. AB - OBJECTIVE: Over the last 50 years, there has been a surge of interest by both the public and medical practitioners in therapies and disciplines that are not considered part of mainstream medical care. The title given to these is complementary and alternative medicine. Of all these branches, our interest is the increasing use of herbal medicines, traditional medicines (such as Chinese or Indian), homeopathy and "dietary supplements," and the influence they may have on our practice. Our objective was to examine the prevalence and reasons for use of complementary and alternative medicines, the current regulations, and proposed policy changes affecting the licensing of these products. In addition, we highlight some of the problems that have been experienced with herbal and traditional medicines. METHODS: A prospective analysis of herbal and over the counter medicines used by elective plastic surgery patients. RESULTS: Of 100 elective plastic surgery patients undergoing procedures at St Andrew's Centre for Burns and Plastic Surgery, 44% of patients were taking a dietary supplement, herbal, or homeopathic remedy. In none of the patients was this documented in the notes by either the surgeon or anesthetist. CONCLUSIONS: We recommend that clear documentation of the use of nonprescribed medicines becomes part of standard practice and, furthermore, that patients stop all such medications 2 weeks prior to surgery until the efficacy, interactions, and safety profiles are clearly established. PMID- 21625529 TI - Systematic exploitation of multiple receptor conformations for virtual ligand screening. AB - The role of virtual ligand screening in modern drug discovery is to mine large chemical collections and to prioritize for experimental testing a comparatively small and diverse set of compounds with expected activity against a target. Several studies have pointed out that the performance of virtual ligand screening can be improved by taking into account receptor flexibility. Here, we systematically assess how multiple crystallographic receptor conformations, a powerful way of discretely representing protein plasticity, can be exploited in screening protocols to separate binders from non-binders. Our analyses encompass 36 targets of pharmaceutical relevance and are based on actual molecules with reported activity against those targets. The results suggest that an ensemble receptor-based protocol displays a stronger discriminating power between active and inactive molecules as compared to its standard single rigid receptor counterpart. Moreover, such a protocol can be engineered not only to enrich a higher number of active compounds, but also to enhance their chemical diversity. Finally, some clear indications can be gathered on how to select a subset of receptor conformations that is most likely to provide the best performance in a real life scenario. PMID- 21625530 TI - Large-scale analysis of acute ethanol exposure in zebrafish development: a critical time window and resilience. AB - BACKGROUND: In humans, ethanol exposure during pregnancy causes a spectrum of developmental defects (fetal alcohol syndrome or FAS). Individuals vary in phenotypic expression. Zebrafish embryos develop FAS-like features after ethanol exposure. In this study, we ask whether stage-specific effects of ethanol can be identified in the zebrafish, and if so, whether they allow the pinpointing of sensitive developmental mechanisms. We have therefore conducted the first large scale (>1500 embryos) analysis of acute, stage-specific drug effects on zebrafish development, with a large panel of readouts. METHODOLOGY/PRINCIPAL FINDINGS: Zebrafish embryos were raised in 96-well plates. Range-finding indicated that 10% ethanol for 1 h was suitable for an acute exposure regime. High-resolution magic angle spinning proton magnetic resonance spectroscopy showed that this produced a transient pulse of 0.86% concentration of ethanol in the embryo within the chorion. Survivors at 5 days postfertilisation were analysed. Phenotypes ranged from normal (resilient) to severely malformed. Ethanol exposure at early stages caused high mortality (>=88%). At later stages of exposure, mortality declined and malformations developed. Pharyngeal arch hypoplasia and behavioral impairment were most common after prim-6 and prim-16 exposure. By contrast, microphthalmia and growth retardation were stage-independent. CONCLUSIONS: Our findings show that some ethanol effects are strongly stage-dependent. The phenotypes mimic key aspects of FAS including craniofacial abnormality, microphthalmia, growth retardation and behavioral impairment. We also identify a critical time window (prim-6 and prim-16) for ethanol sensitivity. Finally, our identification of a wide phenotypic spectrum is reminiscent of human FAS, and may provide a useful model for studying disease resilience. PMID- 21625531 TI - The indoleamine-2,3-dioxygenase (IDO) inhibitor 1-methyl-D-tryptophan upregulates IDO1 in human cancer cells. AB - 1-methyl-D-tryptophan (1-D-MT) is currently being used in clinical trials in patients with relapsed or refractory solid tumors with the aim of inhibiting indoleamine-2,3-dioxygenase (IDO)-mediated tumor immune escape. IDO is expressed in tumors and tumor-draining lymph nodes and degrades tryptophan (trp) to create an immunsuppressive micromilieu both by depleting trp and by accumulating immunosuppressive metabolites of the kynurenine (kyn) pathway. Here we show that proliferation of alloreactive T-cells cocultured with IDO1-positive human cancer cells paradoxically was inhibited by 1-D-MT. Surprisingly incubation with 1-D-MT increased kyn production of human cancer cells. Cell-free assays revealed that 1 D-MT did not alter IDO1 enzymatic activity. Instead, 1-D-MT induced IDO1 mRNA and protein expression through pathways involving p38 MAPK and JNK signalling. Treatment of cancer patients with 1-D-MT has transcriptional effects that may promote rather than suppress anti-tumor immune escape by increasing IDO1 in the cancer cells. These off-target effects should be carefully analyzed in the ongoing clinical trials with 1-D-MT. PMID- 21625532 TI - The sensory consequences of speaking: parametric neural cancellation during speech in auditory cortex. AB - When we speak, we provide ourselves with auditory speech input. Efficient monitoring of speech is often hypothesized to depend on matching the predicted sensory consequences from internal motor commands (forward model) with actual sensory feedback. In this paper we tested the forward model hypothesis using functional Magnetic Resonance Imaging. We administered an overt picture naming task in which we parametrically reduced the quality of verbal feedback by noise masking. Presentation of the same auditory input in the absence of overt speech served as listening control condition. Our results suggest that a match between predicted and actual sensory feedback results in inhibition of cancellation of auditory activity because speaking with normal unmasked feedback reduced activity in the auditory cortex compared to listening control conditions. Moreover, during self-generated speech, activation in auditory cortex increased as the feedback quality of the self-generated speech decreased. We conclude that during speaking early auditory cortex is involved in matching external signals with an internally generated model or prediction of sensory consequences, the locus of which may reside in auditory or higher order brain areas. Matching at early auditory cortex may provide a very sensitive monitoring mechanism that highlights speech production errors at very early levels of processing and may efficiently determine the self-agency of speech input. PMID- 21625533 TI - The SOS pilot study: a RCT of routine oxygen supplementation early after acute stroke--effect on recovery of neurological function at one week. AB - Mild hypoxia is common after stroke and associated with poor long-term outcome. Oxygen supplementation could prevent hypoxia and improve recovery. A previous study of routine oxygen supplementation showed no significant benefit at 7 and 12 months. This pilot study reports the effects of routine oxygen supplementation for 72 hours on oxygen saturation and neurological outcomes at 1 week after a stroke. METHODS: Patients with a clinical diagnosis of acute stroke were recruited within 24 h of hospital admission between October 2004 and April 2008. Participants were randomized to oxygen via nasal cannulae (72 h) or control (room air, oxygen given only if clinically indicated). Clinical outcomes were assessed by research team members at 1 week. Baseline data for oxygen (n = 148) and control (n = 141) did not differ between groups. RESULTS: The median (interquartile range) National Institutes of Health Stroke Scale (NIHSS) score for the groups at baseline was 6 (7) and 5 (7) respectively. The median Nocturnal Oxygen Saturation during treatment was 1.4% (0.3) higher in the oxygen than in the control group (p<0.001) during the intervention. At 1 week, the median NIHSS score had reduced by 2 (3) in the oxygen and by 1 (2) in the control group. 31% of participants in the oxygen group and 14% in the control group had an improvement of >=4 NIHSS points at 1 week doubling the odds of improvement in the oxygen group (OR: 2.9). CONCLUSION: Our data show that routine oxygen supplementation started within 24 hours of hospital admission with acute stroke led to a small, but statistically significant, improvement in neurological recovery at 1 week. However, the difference in NIHSS improvement may be due to baseline imbalance in stroke severity between the two groups and needs to be confirmed in a larger study and linked to longer-term clinical outcome. TRIAL REGISTRATION: Controlled-Trials.com ISRCTN12362720; European Clinical Trials Database 2004-001866-41. PMID- 21625534 TI - AAV exploits subcellular stress associated with inflammation, endoplasmic reticulum expansion, and misfolded proteins in models of cystic fibrosis. AB - Barriers to infection act at multiple levels to prevent viruses, bacteria, and parasites from commandeering host cells for their own purposes. An intriguing hypothesis is that if a cell experiences stress, such as that elicited by inflammation, endoplasmic reticulum (ER) expansion, or misfolded proteins, then subcellular barriers will be less effective at preventing viral infection. Here we have used models of cystic fibrosis (CF) to test whether subcellular stress increases susceptibility to adeno-associated virus (AAV) infection. In human airway epithelium cultured at an air/liquid interface, physiological conditions of subcellular stress and ER expansion were mimicked using supernatant from mucopurulent material derived from CF lungs. Using this inflammatory stimulus to recapitulate stress found in diseased airways, we demonstrated that AAV infection was significantly enhanced. Since over 90% of CF cases are associated with a misfolded variant of Cystic Fibrosis Transmembrane Conductance Regulator (DeltaF508-CFTR), we then explored whether the presence of misfolded proteins could independently increase susceptibility to AAV infection. In these models, AAV was an order of magnitude more efficient at transducing cells expressing DeltaF508-CFTR than in cells expressing wild-type CFTR. Rescue of misfolded DeltaF508-CFTR under low temperature conditions restored viral transduction efficiency to that demonstrated in controls, suggesting effects related to protein misfolding were responsible for increasing susceptibility to infection. By testing other CFTR mutants, G551D, D572N, and 1410X, we have shown this phenomenon is common to other misfolded proteins and not related to loss of CFTR activity. The presence of misfolded proteins did not affect cell surface attachment of virus or influence expression levels from promoter transgene cassettes in plasmid transfection studies, indicating exploitation occurs at the level of virion trafficking or processing. Thus, we surmised that factors enlisted to process misfolded proteins such as DeltaF508-CFTR in the secretory pathway also act to restrict viral infection. In line with this hypothesis, we found that AAV trafficked to the microtubule organizing center and localized near Golgi/ER transport proteins. Moreover, AAV infection efficiency could be modulated with siRNA-mediated knockdown of proteins involved in processing DeltaF508-CFTR or sorting retrograde cargo from the Golgi and ER (calnexin, KDEL R, beta-COP, and PSMB3). In summary, our data support a model where AAV exploits a compromised secretory system and, importantly, underscore the gravity with which a stressed subcellular environment, under internal or external insults, can impact infection efficiency. PMID- 21625535 TI - Mitochondrial ubiquitin ligase MARCH5 promotes TLR7 signaling by attenuating TANK action. AB - The signaling of Toll-like receptors (TLRs) is the host's first line of defense against microbial invasion. The mitochondrion is emerging as a critical platform for antiviral signal transduction. The regulatory role of mitochondria for TLR signaling remains to be explored. Here, we show that the mitochondrial outer membrane protein MARCH5 positively regulates TLR7 signaling. Ectopic expression or knockdown of MARCH5 enhances or impairs NF-kappaB-mediated gene expression, respectively. MARCH5 interacts specifically with TANK, and this interaction is enhanced by R837 stimulation. MARCH5 catalyzes the K63-linked poly-ubiquitination of TANK on its Lysines 229, 233, 280, 302 and 306, thus impairing the ability of TANK to inhibit TRAF6. Mislocalization of MARCH5 abolishes its action on TANK, revealing the critical role of mitochondria in modulating innate immunity. Arguably, this represents the first study linking mitochondria to TLR signaling. PMID- 21625536 TI - Reactive oxygen species hydrogen peroxide mediates Kaposi's sarcoma-associated herpesvirus reactivation from latency. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) establishes a latent infection in the host following an acute infection. Reactivation from latency contributes to the development of KSHV-induced malignancies, which include Kaposi's sarcoma (KS), the most common cancer in untreated AIDS patients, primary effusion lymphoma and multicentric Castleman's disease. However, the physiological cues that trigger KSHV reactivation remain unclear. Here, we show that the reactive oxygen species (ROS) hydrogen peroxide (H2O2) induces KSHV reactivation from latency through both autocrine and paracrine signaling. Furthermore, KSHV spontaneous lytic replication, and KSHV reactivation from latency induced by oxidative stress, hypoxia, and proinflammatory and proangiogenic cytokines are mediated by H2O2. Mechanistically, H2O2 induction of KSHV reactivation depends on the activation of mitogen-activated protein kinase ERK1/2, JNK, and p38 pathways. Significantly, H2O2 scavengers N-acetyl-L-cysteine (NAC), catalase and glutathione inhibit KSHV lytic replication in culture. In a mouse model of KSHV induced lymphoma, NAC effectively inhibits KSHV lytic replication and significantly prolongs the lifespan of the mice. These results directly relate KSHV reactivation to oxidative stress and inflammation, which are physiological hallmarks of KS patients. The discovery of this novel mechanism of KSHV reactivation indicates that antioxidants and anti-inflammation drugs could be promising preventive and therapeutic agents for effectively targeting KSHV replication and KSHV-related malignancies. PMID- 21625537 TI - Amyloid-associated nucleic acid hybridisation. AB - Nucleic acids promote amyloid formation in diseases including Alzheimer's and Creutzfeldt-Jakob disease. However, it remains unclear whether the close interactions between amyloid and nucleic acid allow nucleic acid secondary structure to play a role in modulating amyloid structure and function. Here we have used a simplified system of short basic peptides with alternating hydrophobic and hydrophilic amino acid residues to study nucleic acid - amyloid interactions. Employing biophysical techniques including X-ray fibre diffraction, circular dichroism spectroscopy and electron microscopy we show that the polymerized charges of nucleic acids concentrate and enhance the formation of amyloid from short basic peptides, many of which would not otherwise form fibres. In turn, the amyloid component binds nucleic acids and promotes their hybridisation at concentrations below their solution K(d), as shown by time resolved FRET studies. The self-reinforcing interactions between peptides and nucleic acids lead to the formation of amyloid nucleic acid (ANA) fibres whose properties are distinct from their component polymers. In addition to their importance in disease and potential in engineering, ANA fibres formed from prebiotically-produced peptides and nucleic acids may have played a role in early evolution, constituting the first entities subject to Darwinian evolution. PMID- 21625538 TI - Septation of infectious hyphae is critical for appressoria formation and virulence in the smut fungus Ustilago maydis. AB - Differentiation of hyphae into specialized infection structures, known as appressoria, is a common feature of plant pathogenic fungi that penetrate the plant cuticle. Appressorium formation in U. maydis is triggered by environmental signals but the molecular mechanism of this hyphal differentiation is largely unknown. Infectious hyphae grow on the leaf surface by inserting regularly spaced retraction septa at the distal end of the tip cell leaving empty sections of collapsed hyphae behind. Here we show that formation of retraction septa is critical for appressorium formation and virulence in U. maydis. We demonstrate that the diaphanous-related formin Drf1 is necessary for actomyosin ring formation during septation of infectious hyphae. Drf1 acts as an effector of a Cdc42 GTPase signaling module, which also consists of the Cdc42-specific guanine nucleotide exchange factor Don1 and the Ste20-like kinase Don3. Deletion of drf1, don1 or don3 abolished formation of retraction septa resulting in reduced virulence. Appressorium formation in these mutants was not completely blocked but infection structures were found only at the tip of short filaments indicating that retraction septa are necessary for appressorium formation in extended infectious hyphae. In addition, appressoria of drf1 mutants penetrated the plant tissue less frequently. PMID- 21625539 TI - The OXI1 kinase pathway mediates Piriformospora indica-induced growth promotion in Arabidopsis. AB - Piriformospora indica is an endophytic fungus that colonizes roots of many plant species and promotes growth and resistance to certain plant pathogens. Despite its potential use in agriculture, little is known on the molecular basis of this beneficial plant-fungal interaction. In a genetic screen for plants, which do not show a P. indica- induced growth response, we isolated an Arabidopsis mutant in the OXI1 (Oxidative Signal Inducible1) gene. OXI1 has been characterized as a protein kinase which plays a role in pathogen response and is regulated by H2O2 and PDK1 (3-PHOSPHOINOSITIDE-DEPENDENT PROTEIN KINASE1). A genetic analysis showed that double mutants of the two closely related PDK1.1 and PDK1.2 genes are defective in the growth response to P. indica. While OXI1 and PDK1 gene expression is upregulated in P. indica-colonized roots, defense genes are downregulated, indicating that the fungus suppresses plant defense reactions. PDK1 is activated by phosphatidic acid (PA) and P. indica triggers PA synthesis in Arabidopsis plants. Under beneficial co-cultivation conditions, H2O2 formation is even reduced by the fungus. Importantly, phospholipase D (PLD)alpha1 or PLDdelta mutants, which are impaired in PA synthesis do not show growth promotion in response to fungal infection. These data establish that the P. indica stimulated growth response is mediated by a pathway consisting of the PLD-PDK1 OXI1 cascade. PMID- 21625540 TI - Co-chaperone HSJ1a dually regulates the proteasomal degradation of ataxin-3. AB - Homo sapiens J domain protein (HSJ1) is a J-domain containing co-chaperone that is known to stimulate ATPase activity of HSP70 chaperone, while it also harbors two ubiquitin (Ub)-interacting motifs (UIMs) that may bind with ubiquitinated substrates and potentially function in protein degradation. We studied the effects of HSJ1a on the protein levels of both normal and the disease--related polyQ-expanded forms of ataxin-3 (Atx3) in cells. The results demonstrate that the N-terminal J-domain and the C-terminal UIM domain of HSJ1a exert opposite functions in regulating the protein level of cellular overexpressed Atx3. This dual regulation is dependent on the binding of the J-domain with HSP70, and the UIM domain with polyUb chains. The J-domain down-regulates the protein level of Atx3 through HSP70 mediated proteasomal degradation, while the UIM domain may alleviate this process via maintaining the ubiquitinated Atx3. We propose that co chaperone HSJ1a orchestrates the balance of substrates in stressed cells in a Yin Yang manner. PMID- 21625541 TI - Severe and persistent depletion of circulating plasmacytoid dendritic cells in patients with 2009 pandemic H1N1 infection. AB - BACKGROUND: Dysregulation of host immune responses plays a critical role in the pathogenesis of severe 2009 pandemic H1N1 infection. Whether H1N1 virus could escape innate immune defense in vivo remains to be investigated. The aim of this study was to evaluate the pattern of innate immune response during human 2009 H1N1 infection. We performed the enumeration of circulating myeloid dendritic cells (mDC) and plasmacytoid DC (pDC) in blood from patients with H1N1 pneumonia shortly after the onset of symptoms and during follow-up at different intervals of time. The analysis of CD4 and CD8 count, CD38 T-cell activation marker and serum cytokine/chemokine plasma levels was also done. METHODOLOGY/PRINCIPAL FINDINGS: Blood samples were collected from 13 hospitalized patients with confirmed H1N1-related pneumonia at time of admission and at weeks 1, 4, and 16 of follow-up. 13 healthy donors were enrolled as controls. In the acute phase of the disease, H1N1-infected patients exhibited a significant depletion in both circulating pDC and mDC in conjunction with a decrease of CD4 and CD8 T cell count. In addition, we found plasmatic hyperproduction of IP-10 and RANTES, whereas increase in T-cell immune activation was found at all time points. When we assessed the changes in DC count over time, we observed a progressive normalization of mDC number. On the contrary, H1N1-infected patients did not achieve a complete recovery of pDC count as values remained lower than healthy controls even after 16 weeks of follow-up. CONCLUSIONS: H1N1 disease is associated with a profound depletion of DC subsets. The persistence of pDC deficit for several weeks after disease recovery could be due to H1N1 virus itself or to a preexisting impairment of innate immunity. PMID- 21625543 TI - Rapid screening of complex DNA samples by single-molecule amplification and sequencing. AB - Microbial cloning makes Sanger sequencing of complex DNA samples possible but is labor intensive. We present a simple, rapid and robust method that enables laboratories without special equipment to perform single-molecule amplicon sequencing, although in a low-throughput manner, from sub-picogram quantities of DNA. The method can also be used for quick quality control of next-generation sequencing libraries, as was demonstrated for a metagenomic sample. PMID- 21625542 TI - Negative associations between corpus callosum midsagittal area and IQ in a representative sample of healthy children and adolescents. AB - Documented associations between corpus callosum size and cognitive ability have heretofore been inconsistent potentially owing to differences in sample characteristics, differing methodologies in measuring CC size, or the use of absolute versus relative measures. We investigated the relationship between CC size and intelligence quotient (IQ) in the NIH MRI Study of Normal Brain Development sample, a large cohort of healthy children and adolescents (aged six to 18, n = 198) recruited to be representative of the US population. CC midsagittal area was measured using an automated system that partitioned the CC into 25 subregions. IQ was measured using the Wechsler Abbreviated Scale of Intelligence (WASI). After correcting for total brain volume and age, a significant negative correlation was found between total CC midsagittal area and IQ (r = -0.147; p = 0.040). Post hoc analyses revealed a significant negative correlation in children (age<12) (r = -0.279; p = 0.004) but not in adolescents (age>=12) (r = -0.005; p = 0.962). Partitioning the subjects by gender revealed a negative correlation in males (r = -0.231; p = 0.034) but not in females (r = 0.083; p = 0.389). Results suggest that the association between CC and intelligence is mostly driven by male children. In children, a significant gender difference was observed for FSIQ and PIQ, and in males, a significant age-group difference was observed for FSIQ and PIQ. These findings suggest that the correlation between CC midsagittal area and IQ may be related to age and gender. PMID- 21625544 TI - Expansion of CD4(+) CD25(+) and CD25(-) T-Bet, GATA-3, Foxp3 and RORgammat cells in allergic inflammation, local lung distribution and chemokine gene expression. AB - Allergic asthma is associated with airway eosinophilia, which is regulated by different T-effector cells. T cells express transcription factors T-bet, GATA-3, RORgammat and Foxp3, representing Th1, Th2, Th17 and Treg cells respectively. No study has directly determined the relative presence of each of these T cell subsets concomitantly in a model of allergic airway inflammation. In this study we determined the degree of expansion of these T cell subsets, in the lungs of allergen challenged mice. Cell proliferation was determined by incorporation of 5 bromo-2'-deoxyuridine (BrdU) together with 7-aminoactnomycin (7-AAD). The immunohistochemical localisation of T cells in the lung microenvironments was also quantified. Local expression of cytokines, chemokines and receptor genes was measured using real-time RT-PCR array analysis in tissue sections isolated by laser microdissection and pressure catapulting technology. Allergen exposure increased the numbers of T-bet(+), GATA-3(+), RORgammat(+) and Foxp3(+) cells in CD4(+)CD25(+) and CD4(+)CD25(-) T cells, with the greatest expansion of GATA-3(+) cells. The majority of CD4(+)CD25(+) T-bet(+), GATA-3(+), RORgammat(+) and Foxp3(+) cells had incorporated BrdU and underwent proliferation during allergen exposure. Allergen exposure led to the accumulation of T-bet(+), GATA-3(+) and Foxp3(+) cells in peribronchial and alveolar tissue, GATA-3(+) and Foxp3(+) cells in perivascular tissue, and RORgammat(+) cells in alveolar tissue. A total of 28 cytokines, chemokines and receptor genes were altered more than 3 fold upon allergen exposure, with expression of half of the genes claimed in all three microenvironments. Our study shows that allergen exposure affects all T effector cells in lung, with a dominant of Th2 cells, but with different local cell distribution, probably due to a distinguished local inflammatory milieu. PMID- 21625545 TI - Outcome assessment of a dedicated HIV positive health care worker clinic at a central hospital in Malawi: a retrospective observational study. AB - BACKGROUND: Malawi has one of the world's lowest densities of Health Care Workers (HCW) per capita. This study evaluates outcomes of a dedicated HCW HIV clinic in Malawi, created at Zomba Central Hospital in January 2007. METHODS AND FINDINGS: Retrospective cohort data was analyzed comparing HCW clinic patient baseline characteristics and treatment outcomes at 18 months after inception, against those attending the general HIV clinic. In-depth interviews and focus group discussions were conducted to explore perceptions of patients and caregivers regarding program value, level of awareness and barriers for uptake amongst HCW. 306 patients were enrolled on antiretroviral therapy (ART) in the HCW HIV clinic, 6784 in the general clinic. Significantly (p<0.01) more HCW clients were initiated on ART on the basis of CD4 as opposed to WHO Stage 3/4 (36% vs.23%). Significantly fewer HCW clients defaulted (6% vs.17%), and died (4% vs.12%). The dedicated HCW HIV clinic was perceived as important and convenient in terms of reduced waiting times, and prompt and high quality care. Improved confidentiality was an appreciated quality of the HCW clinic however barriers included fear of being recognized. CONCLUSIONS/SIGNIFICANCE: Outcomes at the HCW clinic appear better compared to the general HIV clinic. The strategy of dedicated clinics to care for health providers is a means of HIV impact mitigation within human resource constrained health systems in high prevalence settings. PMID- 21625546 TI - The participation of calponin in the cross talk between 20-hydroxyecdysone and juvenile hormone signaling pathways by phosphorylation variation. AB - 20-hydroxyecdysone (20E) and juvenile hormone (JH) signaling pathways interact to mediate insect development, but the mechanism of this interaction is poorly understood. Here, a calponin homologue domain (Chd) containing protein (HaCal) is reported to play a key role in the cross talk between 20E and JH signaling by varying its phosphorylation. Chd is known as an actin binding domain present in many proteins including some signaling proteins. Using an epidermal cell line (HaEpi), HaCal was found to be up-regulated by either 20E or the JH analog methoprene (JHA). 20E induced rapid phosphorylation of HaCal whereas no phosphorylation occurred with JHA. HaCal could be quickly translocated into the nuclei through 20E or JH signaling but interacted with USP1 only under the mediation of JHA. Knockdown of HaCal by RNAi blocked the 20E inducibility of USP1, PKC and HR3, and also blocked the JHA inducibility of USP1, PKC and JHi. After gene silencing of HaCal by ingestion of dsHaCal expressed by Escherichia coli, the larval development was arrested and the gene expression of USP1, PKC, HR3 and JHi were blocked. These composite data suggest that HaCal plays roles in hormonal signaling by quickly transferring into nucleus to function as a phosphorylated form in the 20E pathway and as a non-phosphorylated form interacting with USP1 in the JH pathway to facilitate 20E or JH signaling cascade, in short, by switching its phosphorylation status to regulate insect development. PMID- 21625547 TI - Circulating KCNH2 current-activating factor in patients with heart failure and ventricular tachyarrhythmia. AB - BACKGROUND: It is estimated that approximately half of the deaths in patients with HF are sudden and that the most likely causes of sudden death are lethal ventricular tachyarrhythmias such as ventricular tachycardia (VT) or fibrillation (VF). However, the precise mechanism of ventricular tachyarrhythmias remains unknown. The KCNH2 channel conducting the delayed rectifier K(+) current (I(Kr)) is recognized as the most susceptible channel in acquired long QT syndrome. Recent findings have revealed that not only suppression but also enhancement of I(Kr) increase vulnerability to major arrhythmic events, as seen in short QT syndrome. Therefore, we investigated the existence of a circulating KCNH2 current modifying factor in patients with HF. METHODOLOGY/PRINCIPAL FINDINGS: We examined the effects of serum of HF patients on recombinant I(Kr) recorded from HEK 293 cells stably expressing KCNH2 by using the whole-cell patch-clamp technique. Study subjects were 14 patients with non-ischemic HF and 6 normal controls. Seven patients had a history of documented ventricular tachyarrhythmias (VT: 7 and VF: 1). Overnight treatment with 2% serum obtained from HF patients with ventricular arrhythmia resulted in a significant enhancement in the peaks of I(Kr) tail currents compared to the serum from normal controls and HF patients without ventricular arrhythmia. CONCLUSIONS/SIGNIFICANCE: Here we provide the first evidence for the presence of a circulating KCNH2 channel activator in patients with HF and ventricular tachyarrhythmias. This factor may be responsible for arhythmogenesis in patients with HF. PMID- 21625548 TI - Differential regulation of PDE5 expression in left and right ventricles of feline hypertrophy models. AB - BACKGROUND: Though long known to affect smooth muscle biology, recent studies indicate that phosphodiesterase 5 (PDE5) is also expressed in myocardium. Recognizing that the regulation of PDE5 in hypertrophy is not well understood, we assessed the response of PDE5 expression and the level of cGMP-dependent kinase I (cGKI) in the left and right ventricles of feline hypertrophy models. METHODOLOGY/PRINCIPAL FINDINGS: Using a cDNA library of feline aortic smooth muscle cells, we identified and cloned PDE5 cDNA for the first time in this species. The sequence shares 98% identity with its human orthologue at the amino acid level. E. coli expression of the cloned allele allowed selection of antibodies with appropriate specificity, facilitating the analysis of PDE5 expression in feline models created by selective proximal aortic (Ao) or pulmonary artery (PA) banding that resulted in hypertrophy of the left ventricle (LV) and right ventricle (RV), respectively. We demonstrated that PDE5 expression responded differentially with a decreased expression in the LV and an increased expression in the RV in the Ao-banded model. Similarly, in the PA-banded model, LV showed reduced expression while the RV expression was unaltered. In addition, the expression of cGKI was significantly decreased in the RV of Ao-banded group, correlating inversely with the increase in PDE5 expression. CONCLUSIONS/SIGNIFICANCE: The differential regulation of PDE5 and cGKI expression suggests that the mechanisms involved in hypertrophy could be different in RV vs. LV. Reciprocal PDE5 and cGKI expression in the RV of Ao-banded model suggests functional significance for PDE5 up-regulation. PMID- 21625549 TI - Ethnic differences in body composition and obesity related risk factors: study in Chinese and white males living in China. AB - The purpose of this cross-sectional observational study was to identify ethnic differences in body composition and obesity-related risk factors between Chinese and white males living in China. 115 Chinese and 114 white male pilots aged 28-63 years were recruited. Fasting body weight, height and blood pressure were measured following standard procedures. Whole-body and segmental body composition were measured using an 8-contact electrode bioimpedance analysis (BIA) system. Fasting serum glucose, fasting plasma total cholesterol (TC), high-density lipoprotein (HDL) cholesterol, and triglycerides (TG) were assessed using automatic biochemistry analyzer. After adjusting for age and body mass index (BMI), Chinese males had significantly higher percentage of body fat (PBF) both with respect to whole body (Chinese: 23.7%+/-0.2% vs. Whites: 22.4%+/-0.2%) and the trunk area (Chinese: 25.0%+/-0.3% vs. Whites: 23.2%+/-0.3%) compared to their white counterparts. At all BMIs, Chinese males had significantly higher fasting glucose levels (Chinese: 5.7+/-1.0 mmol/L vs. Whites: 5.2+/-1.0 mmol/L) but lower high-density lipoprotein levels (Chinese: 0.8+/-1.0 mmol/L vs. Whites: 1.0+/-1.0 mmol/L) than white males. In addition, a marginally significantly higher diastolic blood pressure was found among Chinese men than that among white men (Chinese: 80+/-1.0 mmHg vs. Whites: 77+/-1.0 mmHg). Chinese males had more body fat and a greater degree of central fat deposition pattern than that seen in white males in the present study. Furthermore, data on blood pressure, fasting glucose and blood lipids suggest that Chinese men may be more prone to obesity related risk factors than white men. PMID- 21625550 TI - Lycopene inhibits NF-kB-mediated IL-8 expression and changes redox and PPARgamma signalling in cigarette smoke-stimulated macrophages. AB - Increasing evidence suggests that lycopene, the major carotenoid present in tomato, may be preventive against smoke-induced cell damage. However, the mechanisms of such a prevention are still unclear. The aim of this study was to investigate the role of lycopene on the production of the pro-inflammatory cytokine IL-8 induced by cigarette smoke and the possible mechanisms implicated. Therefore, human THP-1 macrophages were exposed to cigarette smoke extract (CSE), alone and following a 6-h pre-treatment with lycopene (0.5-2 uM). CSE enhanced IL 8 production in a time- and a dose-dependent manner. Lycopene pre-treatment resulted in a significant inhibition of CSE-induced IL-8 expression at both mRNA and protein levels. NF-kB controlled the transcription of IL-8 induced by CSE, since PDTC prevented such a production. Lycopene suppressed CSE-induced NF-kB DNA binding, NF-kB/p65 nuclear translocation and phosphorylation of IKKalpha and IkBalpha. Such an inhibition was accompanied by a decrease in CSE-induced ROS production and NOX-4 expression. Lycopene further inhibited CSE-induced phosphorylation of the redox-sensitive ERK1/2, JNK and p38 MAPKs. Moreover, the carotenoid increased PPARgamma levels which, in turn, enhanced PTEN expression and decreased pAKT levels in CSE-exposed cells. Such effects were abolished by the PPARgamma inhibitor GW9662. Taken together, our data indicate that lycopene prevented CSE-induced IL-8 production through a mechanism involving an inactivation of NF-kB. NF-kB inactivation was accompanied by an inhibition of redox signalling and an activation of PPARgamma signalling. The ability of lycopene in inhibiting IL-8 production, NF-kB/p65 nuclear translocation, and redox signalling and in increasing PPARgamma expression was also found in isolated rat alveolar macrophages exposed to CSE. These findings provide novel data on new molecular mechanisms by which lycopene regulates cigarette smoke driven inflammation in human macrophages. PMID- 21625551 TI - Trade-off between toxicity and signal detection orchestrated by frequency- and density-dependent genes. AB - Behaviors in insects are partly highly efficient Bayesian processes that fulfill exploratory tasks ending with the colonization of new ecological niches. The foraging (for) gene in Drosophila encodes a cGMP-dependent protein kinase (PKG). It has been extensively described as a frequency-dependent gene and its transcripts are differentially expressed between individuals, reflecting the population density context. Some for transcripts, when expressed in a population at high density for many generations, concomitantly trigger strong dispersive behavior associated with foraging activity. Moreover, genotype-by-environment interaction (GEI) analysis has highlighted a dormant role of for in energetic metabolism in a food deprivation context. In our current report, we show that alleles of for encoding different cGMP-dependent kinase isoforms influence the oxidation of aldehyde groups of aromatic molecules emitted by plants via Aldh-III and a phosphorylatable adaptor. The enhanced efficiency of oxidation of aldehyde odorants into carboxyl groups by the action of for lessens their action and toxicity, which should facilitate exploration and guidance in a complex odor environment. Our present data provide evidence that optimal foraging performance requires the fast metabolism of volatile compounds emitted by plants to avoid neurosensory saturation and that the frequency-dependent genes that trigger dispersion influence these processes. PMID- 21625552 TI - Assembly of designer TAL effectors by Golden Gate cloning. AB - Generation of customized DNA binding domains targeting unique sequences in complex genomes is crucial for many biotechnological applications. The recently described DNA binding domain of the transcription activator-like effectors (TALEs) from Xanthomonas consists of a series of repeats arranged in tandem, each repeat binding a nucleotide of the target sequence. We present here a strategy for engineering of TALE proteins with novel DNA binding specificities based on the 17.5 repeat-containing AvrBs3 TALE as a scaffold. For each of the 17 full repeats, four module types were generated, each with a distinct base preference. Using this set of 68 repeat modules, recognition domains for any 17 nucleotide DNA target sequence of choice can be constructed by assembling selected modules in a defined linear order. Assembly is performed in two successive one-pot cloning steps using the Golden Gate cloning method that allows seamless fusion of multiple DNA fragments. Applying this strategy, we assembled designer TALEs with new target specificities and tested their function in vivo. PMID- 21625553 TI - Global biodiversity and phylogenetic evaluation of remipedia (crustacea). AB - Remipedia is one of the most recently discovered classes of crustaceans, first described in 1981 from anchialine caves in the Bahamas Archipelago. The class is divided into the order Enantiopoda, represented by two fossil species, and Nectiopoda, which contains all known extant remipedes. Since their discovery, the number of nectiopodan species has increased to 24, half of which were described during the last decade. Nectiopoda exhibit a disjunct global distribution pattern, with the highest abundance and diversity in the Caribbean region, and isolated species in the Canary Islands and in Western Australia. Our review of Remipedia provides an overview of their ecological characteristics, including a detailed list of all anchialine marine caves, from which species have been recorded. We discuss alternative hypotheses of the phylogenetic position of Remipedia within Arthropoda, and present first results of an ongoing molecular phylogenetic analysis that do not support the monophyly of several nectiopodan taxa. We believe that a taxonomic revision of Remipedia is absolutely essential, and that a comprehensive revision should include a reappraisal of the fossil record. PMID- 21625554 TI - A gene-phenotype network for the laboratory mouse and its implications for systematic phenotyping. AB - The laboratory mouse is the pre-eminent model organism for the dissection of human disease pathways. With the advent of a comprehensive panel of gene knockouts, projects to characterise the phenotypes of all knockout lines are being initiated. The range of genotype-phenotype associations can be represented using the Mammalian Phenotype ontology. Using publicly available data annotated with this ontology we have constructed gene and phenotype networks representing these associations. These networks show a scale-free, hierarchical and modular character and community structure. They also exhibit enrichment for gene coexpression, protein-protein interactions and Gene Ontology annotation similarity. Close association between gene communities and some high-level ontology terms suggests that systematic phenotyping can provide a direct insight into underlying pathways. However some phenotypes are distributed more diffusely across gene networks, likely reflecting the pleiotropic roles of many genes. Phenotype communities show a many-to-many relationship to human disease communities, but stronger overlap at more granular levels of description. This may suggest that systematic phenotyping projects should aim for high granularity annotations to maximise their relevance to human disease. PMID- 21625555 TI - Modeling a new water channel that allows SET9 to dimethylate p53. AB - SET9, a protein lysine methyltransferase, has been thought to be capable of transferring only one methyl group to target lysine residues. However, some reports have pointed out that SET9 can dimethylate Lys372 of p53 (p53-K372) and Lys4 of histone H3 (H3-K4). In order to understand how p53 can be dimethylated by SET9, we measured the radius of the channel that surrounds p53-K372, first on the basis of the crystal structure of SET9, and we show that the channel is not suitable for water movement. Second, molecular dynamic (MD) simulations were carried out for 204 ns on the crystal structure of SET9. The results show that water leaves the active site of SET9 through a new channel, which is made of G292, A295, Y305 and Y335. In addition, the results of molecular docking and MD simulations indicate that the new water channel continues to remain open when S adenosyl-L-methionine (AdoMet) or S-adenosyl-L-homocysteine (AdoHcy) is bound to SET9. The changes in the radii of these two channels were measured in the equilibrium phase at the constant temperature of 300 K. The results indicate that the first channel still does not allow water to get into or out of the active site, but the new channel is large enough to allow this water to circulate. Our results indicate that water can be removed from the active site, an essential process for allowing the dimethylation reaction to occur. PMID- 21625556 TI - New insights into placozoan sexual reproduction and development. AB - Unraveling animal life cycles and embryonic development is basic to understanding animal biology and often sheds light on phylogenetic relationships. A key group for understanding the evolution of the Metazoa is the early branching phylum Placozoa, which has attracted rapidly increasing attention. Despite over a hundred years of placozoan research the life cycle of this enigmatic phylum remains unknown. Placozoa are a unique model system for which the nuclear genome was published before the basic biology (i.e. life cycle and development) has been unraveled. Four organismal studies have reported the development of oocytes and one genetic study has nourished the hypothesis of sexual reproduction in natural populations at least in the past. Here we report new observations on sexual reproduction and embryonic development in the Placozoa and support the hypothesis of current sexual reproduction. The regular observation of oocytes and expressed sperm markers provide support that placozoans reproduce sexually in the field. Using whole genome and EST sequences and additional cDNA cloning we identified five conserved sperm markers, characteristic for different stages in spermatogenesis. We also report details on the embryonic development up to a 128 cell stage and new ultrastructural features occurring during early development. These results suggest that sperm and oocyte generation and maturation occur in different placozoans and that clonal lineages reproduce bisexually in addition to the standard mode of vegetative reproduction. The sum of observations is best congruent with the hypothesis of a simple life cycle with an alternation of reproductive modes between bisexual and vegetative reproduction. PMID- 21625557 TI - Better few than hungry: flexible feeding ecology of collared lemurs Eulemur collaris in littoral forest fragments. AB - BACKGROUND: Frugivorous primates are known to encounter many problems to cope with habitat degradation, due to the fluctuating spatial and temporal distribution of their food resources. Since lemur communities evolved strategies to deal with periods of food scarcity, these primates are expected to be naturally adapted to fluctuating ecological conditions and to tolerate a certain degree of habitat changes. However, behavioral and ecological strategies adopted by frugivorous lemurs to survive in secondary habitats have been little investigated. Here, we compared the behavioral ecology of collared lemurs (Eulemur collaris) in a degraded fragment of littoral forest of south-east Madagascar, Mandena, with that of their conspecifics in a more intact habitat, Sainte Luce. METHODOLOGY/PRINCIPAL FINDINGS: Lemur groups in Mandena and in Sainte Luce were censused in 2004/2007 and in 2000, respectively. Data were collected via instantaneous sampling on five lemur groups totaling 1,698 observation hours. The Shannon index was used to determine dietary diversity and nutritional analyses were conducted to assess food quality. All feeding trees were identified and measured, and ranging areas determined via the minimum convex polygon. In the degraded area lemurs were able to modify several aspects of their feeding strategies by decreasing group size and by increasing feeding time, ranging areas, and number of feeding trees. The above strategies were apparently able to counteract a clear reduction in both food quality and size of feeding trees. CONCLUSIONS/SIGNIFICANCE: Our findings indicate that collared lemurs in littoral forest fragments modified their behavior to cope with the pressures of fluctuating resource availability. The observed flexibility is likely to be an adaptation to Malagasy rainforests, which are known to undergo periods of fruit scarcity and low productivity. These results should be carefully considered when relocating lemurs or when selecting suitable areas for their conservation. PMID- 21625558 TI - Do epigeal termite mounds increase the diversity of plant habitats in a tropical rain forest in peninsular Malaysia? AB - The extent to which environmental heterogeneity can account for tree species coexistence in diverse ecosystems, such as tropical rainforests, is hotly debated, although the importance of spatial variability in contributing to species co-existence is well recognized. Termites contribute to the micro topographical and nutrient spatial heterogeneity of tropical forests. We therefore investigated whether epigeal termite mounds could contribute to the coexistence of plant species within a 50 ha plot at Pasoh Forest Reserve, Malaysia. Overall, stem density was significantly higher on mounds than in their immediate surroundings, but tree species diversity was significantly lower. Canonical correspondence analysis showed that location on or off mounds significantly influenced species distribution when stems were characterized by basal area. Like studies of termite mounds in other ecosystems, our results suggest that epigeal termite mounds provide a specific microhabitat for the enhanced growth and survival of certain species in these species-rich tropical forests. However, the extent to which epigeal termite mounds facilitate species coexistence warrants further investigation. PMID- 21625559 TI - Regulation of zebrafish hatching by tetraspanin cd63. AB - Tetraspanins cause the clustering of membrane proteins into a level of organisation essential for cellular function. Given the importance and complicated nature of this mechanism, we attempted a novel approach to identify the function of a single component in a biologically relevant context. A morpholino knockdown strategy was used to investigate the role of cd63, a membrane protein associated with intracellular transport and a melanoma marker, in embryonic zebrafish. By using three separate morpholinos targeting cd63, we were able to identify a specific phenotype. Strikingly, morphant fish failed to hatch due to the lack of secreted proteolytic enzymes required for chorion softening. The morphology of the hatching gland at both the cellular and intracellular levels was disorganised, suggesting a role for cd63 in the functioning of this organ. This work identifies a specific role for cd63 in the zebrafish embryo and provides evidence for the suitability of zebrafish as a model system for the investigation of tetraspanin enriched microdomains. PMID- 21625560 TI - Crystal structure of the PAC1R extracellular domain unifies a consensus fold for hormone recognition by class B G-protein coupled receptors. AB - Pituitary adenylate cyclase activating polypeptide (PACAP) is a member of the PACAP/glucagon family of peptide hormones, which controls many physiological functions in the immune, nervous, endocrine, and muscular systems. It activates adenylate cyclase by binding to its receptor, PAC1R, a member of class B G protein coupled receptors (GPCR). Crystal structures of a number of Class B GPCR extracellular domains (ECD) bound to their respective peptide hormones have revealed a consensus mechanism of hormone binding. However, the mechanism of how PACAP binds to its receptor remains controversial as an NMR structure of the PAC1R ECD/PACAP complex reveals a different topology of the ECD and a distinct mode of ligand recognition. Here we report a 1.9 A crystal structure of the PAC1R ECD, which adopts the same fold as commonly observed for other members of Class B GPCR. Binding studies and cell-based assays with alanine-scanned peptides and mutated receptor support a model that PAC1R uses the same conserved fold of Class B GPCR ECD for PACAP binding, thus unifying the consensus mechanism of hormone binding for this family of receptors. PMID- 21625561 TI - Contrasting codon usage patterns and purifying selection at the mating locus in putatively asexual alternaria fungal species. AB - Sexual reproduction in heterothallic ascomycete fungi is controlled by a single mating-type locus called MAT1 with two alternate alleles or idiomorphs, MAT1-1 and MAT1-2. These alleles lack sequence similarity and encode different transcriptional regulators. A large number of phytopathogenic fungi including Alternaria spp. are considered asexual, yet still carry expressed MAT1 genes. The molecular evolution of Alternaria MAT1 was explored using nucleotide diversity, nonsynonymous vs. synonymous substitution (dn/ds) ratios and codon usage statistics. Likelihood ratio tests of site-branch models failed to detect positive selection on MAT1-1-1 or MAT1-2-1. Codon-site models demonstrated that both MAT1-1-1 and MAT1-2-1 are under purifying selection and significant differences in codon usage were observed between MAT1-1-1 and MAT1-2-1. Mean GC content at the third position (GC3) and effective codon usage (ENC) were significantly different between MAT1-1-1 and MAT1-2-1 with values of 0.57 and 48 for MAT1-1-1 and 0.62 and 46 for MAT1-2-1, respectively. In contrast, codon usage of Pleospora spp. (anamorph Stemphylium), a closely related Dothideomycete genus, was not significantly different between MAT1-1-1 and MAT1-2-1. The purifying selection and biased codon usage detected at the MAT1 locus in Alternaria spp. suggest a recent sexual past, cryptic sexual present and/or that MAT1 plays important cellular role(s) in addition to mating. PMID- 21625562 TI - Post-embryonic nerve-associated precursors to adult pigment cells: genetic requirements and dynamics of morphogenesis and differentiation. AB - The pigment cells of vertebrates serve a variety of functions and generate a stunning variety of patterns. These cells are also implicated in human pathologies including melanoma. Whereas the events of pigment cell development have been studied extensively in the embryo, much less is known about morphogenesis and differentiation of these cells during post-embryonic stages. Previous studies of zebrafish revealed genetically distinct populations of embryonic and adult melanophores, the ectotherm homologue of amniote melanocytes. Here, we use molecular markers, vital labeling, time-lapse imaging, mutational analyses, and transgenesis to identify peripheral nerves as a niche for precursors to adult melanophores that subsequently migrate to the skin to form the adult pigment pattern. We further identify genetic requirements for establishing, maintaining, and recruiting precursors to the adult melanophore lineage and demonstrate novel compensatory behaviors during pattern regulation in mutant backgrounds. Finally, we show that distinct populations of latent precursors having differential regenerative capabilities persist into the adult. These findings provide a foundation for future studies of post-embryonic pigment cell precursors in development, evolution, and neoplasia. PMID- 21625563 TI - A nervous origin for fish stripes. PMID- 21625564 TI - The stb operon balances the requirements for vegetative stability and conjugative transfer of plasmid R388. AB - The conjugative plasmid R388 and a number of other plasmids carry an operon, stbABC, adjacent to the origin of conjugative transfer. We investigated the role of the stbA, stbB, and stbC genes. Deletion of stbA affected both conjugation and stability. It led to a 50-fold increase in R388 transfer frequency, as well as to high plasmid loss. In contrast, deletion of stbB abolished conjugation but provoked no change in plasmid stability. Deletion of stbC showed no effect, neither in conjugation nor in stability. Deletion of the entire stb operon had no effect on conjugation, which remained as in the wild-type plasmid, but led to a plasmid loss phenotype similar to that of the R388DeltastbA mutant. We concluded that StbA is required for plasmid stability and that StbA and StbB control conjugation. We next observed the intracellular positioning of R388 DNA molecules and showed that they localize as discrete foci evenly distributed in live Escherichia coli cells. Plasmid instability of the R388DeltaDeltastbA mutant correlated with aberrant localization of the plasmid DNA molecules as clusters, either at one cell pole, at both poles, or at the cell center. In contrast, plasmid molecules in the R388DeltaDeltastbB mutant were mostly excluded from the cell poles. Thus, results indicate that defects in both plasmid maintenance and transfer are a consequence of variations in the intracellular positioning of plasmid DNA. We propose that StbA and StbB constitute an atypical plasmid stabilization system that reconciles two modes of plasmid R388 physiology: a maintenance mode (replication and segregation) and a propagation mode (conjugation). The consequences of this novel concept in plasmid physiology will be discussed. PMID- 21625565 TI - deFuse: an algorithm for gene fusion discovery in tumor RNA-Seq data. AB - Gene fusions created by somatic genomic rearrangements are known to play an important role in the onset and development of some cancers, such as lymphomas and sarcomas. RNA-Seq (whole transcriptome shotgun sequencing) is proving to be a useful tool for the discovery of novel gene fusions in cancer transcriptomes. However, algorithmic methods for the discovery of gene fusions using RNA-Seq data remain underdeveloped. We have developed deFuse, a novel computational method for fusion discovery in tumor RNA-Seq data. Unlike existing methods that use only unique best-hit alignments and consider only fusion boundaries at the ends of known exons, deFuse considers all alignments and all possible locations for fusion boundaries. As a result, deFuse is able to identify fusion sequences with demonstrably better sensitivity than previous approaches. To increase the specificity of our approach, we curated a list of 60 true positive and 61 true negative fusion sequences (as confirmed by RT-PCR), and have trained an adaboost classifier on 11 novel features of the sequence data. The resulting classifier has an estimated value of 0.91 for the area under the ROC curve. We have used deFuse to discover gene fusions in 40 ovarian tumor samples, one ovarian cancer cell line, and three sarcoma samples. We report herein the first gene fusions discovered in ovarian cancer. We conclude that gene fusions are not infrequent events in ovarian cancer and that these events have the potential to substantially alter the expression patterns of the genes involved; gene fusions should therefore be considered in efforts to comprehensively characterize the mutational profiles of ovarian cancer transcriptomes. PMID- 21625566 TI - A toolbox model of evolution of metabolic pathways on networks of arbitrary topology. AB - In prokaryotic genomes the number of transcriptional regulators is known to be proportional to the square of the total number of protein-coding genes. A toolbox model of evolution was recently proposed to explain this empirical scaling for metabolic enzymes and their regulators. According to its rules, the metabolic network of an organism evolves by horizontal transfer of pathways from other species. These pathways are part of a larger "universal" network formed by the union of all species-specific networks. It remained to be understood, however, how the topological properties of this universal network influence the scaling law of functional content of genomes in the toolbox model. Here we answer this question by first analyzing the scaling properties of the toolbox model on arbitrary tree-like universal networks. We prove that critical branching topology, in which the average number of upstream neighbors of a node is equal to one, is both necessary and sufficient for quadratic scaling. We further generalize the rules of the model to incorporate reactions with multiple substrates/products as well as branched and cyclic metabolic pathways. To achieve its metabolic tasks, the new model employs evolutionary optimized pathways with minimal number of reactions. Numerical simulations of this realistic model on the universal network of all reactions in the KEGG database produced approximately quadratic scaling between the number of regulated pathways and the size of the metabolic network. To quantify the geometrical structure of individual pathways, we investigated the relationship between their number of reactions, byproducts, intermediate, and feedback metabolites. Our results validate and explain the ubiquitous appearance of the quadratic scaling for a broad spectrum of topologies of underlying universal metabolic networks. They also demonstrate why, in spite of "small-world" topology, real-life metabolic networks are characterized by a broad distribution of pathway lengths and sizes of metabolic regulons in regulatory networks. PMID- 21625567 TI - A decline in p38 MAPK signaling underlies immunosenescence in Caenorhabditis elegans. AB - The decline in immune function with aging, known as immunosenescence, has been implicated in evolutionarily diverse species, but the underlying molecular mechanisms are not understood. During aging in Caenorhabditis elegans, intestinal tissue deterioration and the increased intestinal proliferation of bacteria are observed, but how innate immunity changes during C. elegans aging has not been defined. Here we show that C. elegans exhibits increased susceptibility to bacterial infection with age, and we establish that aging is associated with a decline in the activity of the conserved PMK-1 p38 mitogen-activated protein kinase pathway, which regulates innate immunity in C. elegans. Our data define the phenomenon of innate immunosenescence in C. elegans in terms of the age dependent dynamics of the PMK-1 innate immune signaling pathway, and they suggest that a cycle of intestinal tissue aging, immunosenescence, and bacterial proliferation leads to death in aging C. elegans. PMID- 21625568 TI - SIV Nef proteins recruit the AP-2 complex to antagonize Tetherin and facilitate virion release. AB - Lentiviral Nef proteins have multiple functions and are important for viral pathogenesis. Recently, Nef proteins from many simian immunodefiency viruses were shown to antagonize a cellular antiviral protein, named Tetherin, that blocks release of viral particles from the cell surface. However, the mechanism by which Nef antagonizes Tetherin is unknown. Here, using related Nef proteins that differ in their ability to antagonize Tetherin, we identify three amino-acids in the C terminal domain of Nef that are critical specifically for its ability to antagonize Tetherin. Additionally, divergent Nef proteins bind to the AP-2 clathrin adaptor complex, and we show that residues important for this interaction are required for Tetherin antagonism, downregulation of Tetherin from the cell surface and removal of Tetherin from sites of particle assembly. Accordingly, depletion of AP-2 using RNA interference impairs the ability of Nef to antagonize Tetherin, demonstrating that AP-2 recruitment is required for Nef proteins to counteract this antiviral protein. PMID- 21625569 TI - Spatial learning and action planning in a prefrontal cortical network model. AB - The interplay between hippocampus and prefrontal cortex (PFC) is fundamental to spatial cognition. Complementing hippocampal place coding, prefrontal representations provide more abstract and hierarchically organized memories suitable for decision making. We model a prefrontal network mediating distributed information processing for spatial learning and action planning. Specific connectivity and synaptic adaptation principles shape the recurrent dynamics of the network arranged in cortical minicolumns. We show how the PFC columnar organization is suitable for learning sparse topological-metrical representations from redundant hippocampal inputs. The recurrent nature of the network supports multilevel spatial processing, allowing structural features of the environment to be encoded. An activation diffusion mechanism spreads the neural activity through the column population leading to trajectory planning. The model provides a functional framework for interpreting the activity of PFC neurons recorded during navigation tasks. We illustrate the link from single unit activity to behavioral responses. The results suggest plausible neural mechanisms subserving the cognitive "insight" capability originally attributed to rodents by Tolman & Honzik. Our time course analysis of neural responses shows how the interaction between hippocampus and PFC can yield the encoding of manifold information pertinent to spatial planning, including prospective coding and distance-to-goal correlates. PMID- 21625570 TI - Structural insights into viral determinants of nematode mediated Grapevine fanleaf virus transmission. AB - Many animal and plant viruses rely on vectors for their transmission from host to host. Grapevine fanleaf virus (GFLV), a picorna-like virus from plants, is transmitted specifically by the ectoparasitic nematode Xiphinema index. The icosahedral capsid of GFLV, which consists of 60 identical coat protein subunits (CP), carries the determinants of this specificity. Here, we provide novel insight into GFLV transmission by nematodes through a comparative structural and functional analysis of two GFLV variants. We isolated a mutant GFLV strain (GFLV TD) poorly transmissible by nematodes, and showed that the transmission defect is due to a glycine to aspartate mutation at position 297 (Gly297Asp) in the CP. We next determined the crystal structures of the wild-type GFLV strain F13 at 3.0 A and of GFLV-TD at 2.7 A resolution. The Gly297Asp mutation mapped to an exposed loop at the outer surface of the capsid and did not affect the conformation of the assembled capsid, nor of individual CP molecules. The loop is part of a positively charged pocket that includes a previously identified determinant of transmission. We propose that this pocket is a ligand-binding site with essential function in GFLV transmission by X. index. Our data suggest that perturbation of the electrostatic landscape of this pocket affects the interaction of the virion with specific receptors of the nematode's feeding apparatus, and thereby severely diminishes its transmission efficiency. These data provide a first structural insight into the interactions between a plant virus and a nematode vector. PMID- 21625571 TI - Cellularly-driven differences in network synchronization propensity are differentially modulated by firing frequency. AB - Spatiotemporal pattern formation in neuronal networks depends on the interplay between cellular and network synchronization properties. The neuronal phase response curve (PRC) is an experimentally obtainable measure that characterizes the cellular response to small perturbations, and can serve as an indicator of cellular propensity for synchronization. Two broad classes of PRCs have been identified for neurons: Type I, in which small excitatory perturbations induce only advances in firing, and Type II, in which small excitatory perturbations can induce both advances and delays in firing. Interestingly, neuronal PRCs are usually attenuated with increased spiking frequency, and Type II PRCs typically exhibit a greater attenuation of the phase delay region than of the phase advance region. We found that this phenomenon arises from an interplay between the time constants of active ionic currents and the interspike interval. As a result, excitatory networks consisting of neurons with Type I PRCs responded very differently to frequency modulation compared to excitatory networks composed of neurons with Type II PRCs. Specifically, increased frequency induced a sharp decrease in synchrony of networks of Type II neurons, while frequency increases only minimally affected synchrony in networks of Type I neurons. These results are demonstrated in networks in which both types of neurons were modeled generically with the Morris-Lecar model, as well as in networks consisting of Hodgkin-Huxley-based model cortical pyramidal cells in which simulated effects of acetylcholine changed PRC type. These results are robust to different network structures, synaptic strengths and modes of driving neuronal activity, and they indicate that Type I and Type II excitatory networks may display two distinct modes of processing information. PMID- 21625572 TI - Inhibition of both HIV-1 reverse transcription and gene expression by a cyclic peptide that binds the Tat-transactivating response element (TAR) RNA. AB - The RNA response element TAR plays a critical role in HIV replication by providing a binding site for the recruitment of the viral transactivator protein Tat. Using a structure-guided approach, we have developed a series of conformationally-constrained cyclic peptides that act as structural mimics of the Tat RNA binding region and block Tat-TAR interactions at nanomolar concentrations in vitro. Here we show that these compounds block Tat-dependent transcription in cell-free systems and in cell-based reporter assays. The compounds are also cell permeable, have low toxicity, and inhibit replication of diverse HIV-1 strains, including both CXCR4-tropic and CCR5-tropic primary HIV-1 isolates of the divergent subtypes A, B, C, D and CRF01_AE. In human peripheral blood mononuclear cells, the cyclic peptidomimetic L50 exhibited an IC(50) ~250 nM. Surprisingly, inhibition of LTR-driven HIV-1 transcription could not account for the full antiviral activity. Timed drug-addition experiments revealed that L-50 has a bi phasic inhibition curve with the first phase occurring after HIV-1 entry into the host cell and during the initiation of HIV-1 reverse transcription. The second phase coincides with inhibition of HIV-1 transcription. Reconstituted reverse transcription assays confirm that HIV-1 (-) strand strong stop DNA synthesis is blocked by L50-TAR RNA interactions in-vitro. These findings are consistent with genetic evidence that TAR plays critical roles both during reverse transcription and during HIV gene expression. Our results suggest that antiviral drugs targeting TAR RNA might be highly effective due to a dual inhibitory mechanism. PMID- 21625573 TI - A multiscale approach to characterize the early aggregation steps of the amyloid forming peptide GNNQQNY from the yeast prion sup-35. AB - The self-organization of peptides into amyloidogenic oligomers is one of the key events for a wide range of molecular and degenerative diseases. Atomic-resolution characterization of the mechanisms responsible for the aggregation process and the resulting structures is thus a necessary step to improve our understanding of the determinants of these pathologies. To address this issue, we combine the accelerated sampling properties of replica exchange molecular dynamics simulations based on the OPEP coarse-grained potential with the atomic resolution description of interactions provided by all-atom MD simulations, and investigate the oligomerization process of the GNNQQNY for three system sizes: 3-mers, 12 mers and 20-mers. Results for our integrated simulations show a rich variety of structural arrangements for aggregates of all sizes. Elongated fibril-like structures can form transiently in the 20-mer case, but they are not stable and easily interconvert in more globular and disordered forms. Our extensive characterization of the intermediate structures and their physico-chemical determinants points to a high degree of polymorphism for the GNNQQNY sequence that can be reflected at the macroscopic scale. Detailed mechanisms and structures that underlie amyloid aggregation are also provided. PMID- 21625574 TI - Type I interferon production induced by Streptococcus pyogenes-derived nucleic acids is required for host protection. AB - Streptococcus pyogenes is a Gram-positive human pathogen that is recognized by yet unknown pattern recognition receptors (PRRs). Engagement of these receptor molecules during infection with S. pyogenes, a largely extracellular bacterium with limited capacity for intracellular survival, causes innate immune cells to produce inflammatory mediators such as TNF, but also type I interferon (IFN). Here we show that signaling elicited by type I IFNs is required for successful defense of mice against lethal subcutaneous cellulitis caused by S. pyogenes. Type I IFN signaling was accompanied with reduced neutrophil recruitment to the site of infection. Mechanistic analysis revealed that macrophages and conventional dendritic cells (cDCs) employ different signaling pathways leading to IFN-beta production. Macrophages required IRF3, STING, TBK1 and partially MyD88, whereas in cDCs the IFN-beta production was fully dependent on IRF5 and MyD88. Furthermore, IFN-beta production by macrophages was dependent on the endosomal delivery of streptococcal DNA, while in cDCs streptococcal RNA was identified as the IFN-beta inducer. Despite a role of MyD88 in both cell types, the known IFN-inducing TLRs were individually not required for generation of the IFN-beta response. These results demonstrate that the innate immune system employs several strategies to efficiently recognize S. pyogenes, a pathogenic bacterium that succeeded in avoiding recognition by the standard arsenal of TLRs. PMID- 21625575 TI - Protective efficacy of serially up-ranked subdominant CD8+ T cell epitopes against virus challenges. AB - Immunodominance in T cell responses to complex antigens like viruses is still incompletely understood. Some data indicate that the dominant responses to viruses are not necessarily the most protective, while other data imply that dominant responses are the most important. The issue is of considerable importance to the rational design of vaccines, particularly against variable escaping viruses like human immunodeficiency virus type 1 and hepatitis C virus. Here, we showed that sequential inactivation of dominant epitopes up-ranks the remaining subdominant determinants. Importantly, we demonstrated that subdominant epitopes can induce robust responses and protect against whole viruses if they are allowed at least once in the vaccination regimen to locally or temporally dominate T cell induction. Therefore, refocusing T cell immune responses away from highly variable determinants recognized during natural virus infection towards subdominant, but conserved regions is possible and merits evaluation in humans. PMID- 21625576 TI - Extensive genome-wide variability of human cytomegalovirus in congenitally infected infants. AB - Research has shown that RNA virus populations are highly variable, most likely due to low fidelity replication of RNA genomes. It is generally assumed that populations of DNA viruses will be less complex and show reduced variability when compared to RNA viruses. Here, we describe the use of high throughput sequencing for a genome wide study of viral populations from urine samples of neonates with congenital human cytomegalovirus (HCMV) infections. We show that HCMV intrahost genomic variability, both at the nucleotide and amino acid level, is comparable to many RNA viruses, including HIV. Within intrahost populations, we find evidence of selective sweeps that may have resulted from immune-mediated mechanisms. Similarly, genome wide, population genetic analyses suggest that positive selection has contributed to the divergence of the HCMV species from its most recent ancestor. These data provide evidence that HCMV, a virus with a large dsDNA genome, exists as a complex mixture of genome types in humans and offer insights into the evolution of the virus. PMID- 21625577 TI - Attracting dynamics of frontal cortex ensembles during memory-guided decision making. AB - A common theoretical view is that attractor-like properties of neuronal dynamics underlie cognitive processing. However, although often proposed theoretically, direct experimental support for the convergence of neural activity to stable population patterns as a signature of attracting states has been sparse so far, especially in higher cortical areas. Combining state space reconstruction theorems and statistical learning techniques, we were able to resolve details of anterior cingulate cortex (ACC) multiple single-unit activity (MSUA) ensemble dynamics during a higher cognitive task which were not accessible previously. The approach worked by constructing high-dimensional state spaces from delays of the original single-unit firing rate variables and the interactions among them, which were then statistically analyzed using kernel methods. We observed cognitive epoch-specific neural ensemble states in ACC which were stable across many trials (in the sense of being predictive) and depended on behavioral performance. More interestingly, attracting properties of these cognitively defined ensemble states became apparent in high-dimensional expansions of the MSUA spaces due to a proper unfolding of the neural activity flow, with properties common across different animals. These results therefore suggest that ACC networks may process different subcomponents of higher cognitive tasks by transiting among different attracting states. PMID- 21625578 TI - Topological cluster analysis reveals the systemic organization of the Caenorhabditis elegans connectome. AB - The modular organization of networks of individual neurons interwoven through synapses has not been fully explored due to the incredible complexity of the connectivity architecture. Here we use the modularity-based community detection method for directed, weighted networks to examine hierarchically organized modules in the complete wiring diagram (connectome) of Caenorhabditis elegans (C. elegans) and to investigate their topological properties. Incorporating bilateral symmetry of the network as an important cue for proper cluster assignment, we identified anatomical clusters in the C. elegans connectome, including a body spanning cluster, which correspond to experimentally identified functional circuits. Moreover, the hierarchical organization of the five clusters explains the systemic cooperation (e.g., mechanosensation, chemosensation, and navigation) that occurs among the structurally segregated biological circuits to produce higher-order complex behaviors. PMID- 21625579 TI - Integrative analysis of transgenic alfalfa (Medicago sativa L.) suggests new metabolic control mechanisms for monolignol biosynthesis. AB - The entanglement of lignin polymers with cellulose and hemicellulose in plant cell walls is a major biological barrier to the economically viable production of biofuels from woody biomass. Recent efforts of reducing this recalcitrance with transgenic techniques have been showing promise for ameliorating or even obviating the need for costly pretreatments that are otherwise required to remove lignin from cellulose and hemicelluloses. At the same time, genetic manipulations of lignin biosynthetic enzymes have sometimes yielded unforeseen consequences on lignin composition, thus raising the question of whether the current understanding of the pathway is indeed correct. To address this question systemically, we developed and applied a novel modeling approach that, instead of analyzing the pathway within a single target context, permits a comprehensive, simultaneous investigation of different datasets in wild type and transgenic plants. Specifically, the proposed approach combines static flux-based analysis with a Monte Carlo simulation in which very many randomly chosen sets of parameter values are evaluated against kinetic models of lignin biosynthesis in different stem internodes of wild type and lignin-modified alfalfa plants. In addition to four new postulates that address the reversibility of some key reactions, the modeling effort led to two novel postulates regarding the control of the lignin biosynthetic pathway. The first posits functionally independent pathways toward the synthesis of different lignin monomers, while the second postulate proposes a novel feedforward regulatory mechanism. Subsequent laboratory experiments have identified the signaling molecule salicylic acid as a potential mediator of the postulated control mechanism. Overall, the results demonstrate that mathematical modeling can be a valuable complement to conventional transgenic approaches and that it can provide biological insights that are otherwise difficult to obtain. PMID- 21625580 TI - How structure determines correlations in neuronal networks. AB - Networks are becoming a ubiquitous metaphor for the understanding of complex biological systems, spanning the range between molecular signalling pathways, neural networks in the brain, and interacting species in a food web. In many models, we face an intricate interplay between the topology of the network and the dynamics of the system, which is generally very hard to disentangle. A dynamical feature that has been subject of intense research in various fields are correlations between the noisy activity of nodes in a network. We consider a class of systems, where discrete signals are sent along the links of the network. Such systems are of particular relevance in neuroscience, because they provide models for networks of neurons that use action potentials for communication. We study correlations in dynamic networks with arbitrary topology, assuming linear pulse coupling. With our novel approach, we are able to understand in detail how specific structural motifs affect pairwise correlations. Based on a power series decomposition of the covariance matrix, we describe the conditions under which very indirect interactions will have a pronounced effect on correlations and population dynamics. In random networks, we find that indirect interactions may lead to a broad distribution of activation levels with low average but highly variable correlations. This phenomenon is even more pronounced in networks with distance dependent connectivity. In contrast, networks with highly connected hubs or patchy connections often exhibit strong average correlations. Our results are particularly relevant in view of new experimental techniques that enable the parallel recording of spiking activity from a large number of neurons, an appropriate interpretation of which is hampered by the currently limited understanding of structure-dynamics relations in complex networks. PMID- 21625581 TI - The H-index as a quantitative indicator of the relative impact of human diseases. AB - Assessment of the relative impact of diseases and pathogens is important for agencies and other organizations charged with providing disease surveillance, management and control. It also helps funders of disease-related research to identify the most important areas for investment. Decisions as to which pathogens or diseases to target are often made using complex risk assessment approaches; however, these usually involve evaluating a large number of hazards as it is rarely feasible to conduct an in-depth appraisal of each. Here we propose the use of the H-index (or Hirsch index) as an alternative rapid, repeatable and objective means of assessing pathogen impact. H-index scores for 1,414 human pathogens were obtained from the Institute for Scientific Information's Web of Science (WOS) in July/August 2010. Scores were compared for zoonotic/non zoonotic, and emerging/non-emerging pathogens, and across taxonomic groups. H indices for a subset of pathogens were compared with Disability Adjusted Life Year (DALY) estimates for the diseases they cause. H-indices ranged from 0 to 456, with a median of 11. Emerging pathogens had higher H-indices than non emerging pathogens. Zoonotic pathogens tended to have higher H-indices than human only pathogens, although the opposite was observed for viruses. There was a significant correlation between the DALY of a disease and the H-index of the pathogen(s) that cause it. Therefore, scientific interest, as measured by the H index, appears to be a reflection of the true impact of pathogens. The H-index method can be utilized to set up an objective, repeatable and readily automated system for assessing pathogen or disease impact. PMID- 21625582 TI - Wolbachia infections are virulent and inhibit the human malaria parasite Plasmodium falciparum in Anopheles gambiae. AB - Endosymbiotic Wolbachia bacteria are potent modulators of pathogen infection and transmission in multiple naturally and artificially infected insect species, including important vectors of human pathogens. Anopheles mosquitoes are naturally uninfected with Wolbachia, and stable artificial infections have not yet succeeded in this genus. Recent techniques have enabled establishment of somatic Wolbachia infections in Anopheles. Here, we characterize somatic infections of two diverse Wolbachia strains (wMelPop and wAlbB) in Anopheles gambiae, the major vector of human malaria. After infection, wMelPop disseminates widely in the mosquito, infecting the fat body, head, sensory organs and other tissues but is notably absent from the midgut and ovaries. Wolbachia initially induces the mosquito immune system, coincident with initial clearing of the infection, but then suppresses expression of immune genes, coincident with Wolbachia replication in the mosquito. Both wMelPop and wAlbB significantly inhibit Plasmodium falciparum oocyst levels in the mosquito midgut. Although not virulent in non-bloodfed mosquitoes, wMelPop exhibits a novel phenotype and is extremely virulent for approximately 12-24 hours post-bloodmeal, after which surviving mosquitoes exhibit similar mortality trajectories to control mosquitoes. The data suggest that if stable transinfections act in a similar manner to somatic infections, Wolbachia could potentially be used as part of a strategy to control the Anopheles mosquitoes that transmit malaria. PMID- 21625583 TI - Partial netrin-1 deficiency aggravates acute kidney injury. AB - The netrin family of secreted proteins provides migrational cues in the developing central nervous system. Recently, netrins have also been shown to regulate diverse processes beyond their functions in the brain, incluing the ochrestration of inflammatory events. Particularly netrin-1 has been implicated in dampening hypoxia-induced inflammation. Here, we hypothesized an anti inflammatory role of endogenous netrin-1 in acute kidney injury (AKI). As homozygous deletion of netrin-1 is lethal, we studied mice with partial netrin-1 deletion (Ntn-1(+/-) mice) as a genetic model. In fact, Ntn-1(+/-) mice showed attenuated Ntn-1 levels at baseline and following ischemic AKI. Functional studies of AKI induced by 30 min of renal ischemia and reperfusion revealed enhanced kidney dysfunction in Ntn-1(+/-) mice as assessed by measurements of glomerular filtration, urine flow rate, urine electrolytes, serum creatinine and creatinine clearance. Consistent with these findings, histological studies indicated a more severe degree kidney injury. Similarly, elevations of renal and systemic inflammatory markers were enhanced in mice with partial netrin-1 deficiency. Finally, treatment of Ntn-1(+/-) mice with exogenous netrin-1 restored a normal phenotype during AKI. Taking together, these studies implicate endogenous netrin-1 in attenuating renal inflammation during AKI. PMID- 21625584 TI - Induction of HIV neutralizing antibodies against the MPER of the HIV envelope protein by HA/gp41 chimeric protein-based DNA and VLP vaccines. AB - Several conserved neutralizing epitopes have been identified in the HIV Env protein and among these, the MPER of gp41 has received great attention and is widely recognized as a promising target. However, little success has been achieved in eliciting MPER-specific HIV neutralizing antibodies by a number of different vaccine strategies. We investigated the ability of HA/gp41 chimeric protein-based vaccines, which were designed to enhance the exposure of the MPER in its native conformation, to induce MPER-specific HIV neutralizing antibodies. In characterization of the HA/gp41 chimeric protein, we found that by mutating an unpaired Cys residue (Cys-14) in its HA1 subunit to a Ser residue, the modified chimeric protein HA-C14S/gp41 showed increased reactivity to a conformation sensitive monoclonal antibody against HA and formed more stable trimers in VLPs. On the other hand, HA-C14S/gp41 and HA/gp41 chimeric proteins expressed on the cell surfaces exhibited similar reactivity to monoclonal antibodies 2F5 and 4E10. Immunization of guinea pigs using the HA-C14S/gp41 DNA or VLP vaccines induced antibodies against the HIV gp41 as well as to a peptide corresponding to a segment of MPER at higher levels than immunization by standard HIV VLPs. Further, sera from vaccinated guinea pigs were found to exhibit HIV neutralizing activities. Moreover, sera from guinea pigs vaccinated by HA-C14S/gp41 DNA and VLP vaccines but not the standard HIV VLPs, were found to neutralize HIV pseudovirions containing a SIV-4E10 chimeric Env protein. The virus neutralization could be blocked by a MPER-specific peptide, thus demonstrating induction of MPER-specific HIV neutralizing antibodies by this novel vaccine strategy. These results show that induction of MPER-specific HIV neutralizing antibodies can be achieved through a rationally designed vaccine strategy. PMID- 21625585 TI - Transcriptional activators of human genes with programmable DNA-specificity. AB - TAL (transcription activator-like) effectors are translocated by Xanthomonas bacteria into plant cells where they activate transcription of target genes. DNA target sequence recognition occurs in a unique mode involving a central domain of tandem repeats. Each repeat recognizes a single base pair in a contiguous DNA sequence and a pair of adjacent hypervariable amino acid residues per repeat specifies which base is bound. Rearranging the repeats allows the design of novel TAL proteins with predictable DNA-recognition specificities. TAL protein-based transcriptional activation in plant cells is mediated by a C-terminal activation domain (AD). Here, we created synthetic TAL proteins with designed repeat compositions using a novel modular cloning strategy termed "Golden TAL Technology". Newly programmed TAL proteins were not only functional in plant cells, but also in human cells and activated targeted expression of exogenous as well as endogenous genes. Transcriptional activation in different human cell lines was markedly improved by replacing the TAL-AD with the VP16-AD of herpes simplex virus. The creation of TAL proteins with potentially any desired DNA recognition specificity allows their versatile use in biotechnology. PMID- 21625586 TI - A general model for binary cell fate decision gene circuits with degeneracy: indeterminacy and switch behavior in the absence of cooperativity. AB - BACKGROUND: The gene regulatory circuit motif in which two opposing fate determining transcription factors inhibit each other but activate themselves has been used in mathematical models of binary cell fate decisions in multipotent stem or progenitor cells. This simple circuit can generate multistability and explains the symmetric "poised" precursor state in which both factors are present in the cell at equal amounts as well as the resolution of this indeterminate state as the cell commits to either cell fate characterized by an asymmetric expression pattern of the two factors. This establishes the two alternative stable attractors that represent the two fate options. It has been debated whether cooperativity of molecular interactions is necessary to produce such multistability. PRINCIPAL FINDINGS: Here we take a general modeling approach and argue that this question is not relevant. We show that non-linearity can arise in two distinct models in which no explicit interaction between the two factors is assumed and that distinct chemical reaction kinetic formalisms can lead to the same (generic) dynamical system form. Moreover, we describe a novel type of bifurcation that produces a degenerate steady state that can explain the metastable state of indeterminacy prior to cell fate decision-making and is consistent with biological observations. CONCLUSION: The general model presented here thus offers a novel principle for linking regulatory circuits with the state of indeterminacy characteristic of multipotent (stem) cells. PMID- 21625587 TI - Acyclic identification of aptamers for human alpha-thrombin using over represented libraries and deep sequencing. AB - BACKGROUND: Aptamers are oligonucleotides that bind proteins and other targets with high affinity and selectivity. Twenty years ago elements of natural selection were adapted to in vitro selection in order to distinguish aptamers among randomized sequence libraries. The primary bottleneck in traditional aptamer discovery is multiple cycles of in vitro evolution. METHODOLOGY/PRINCIPAL FINDINGS: We show that over-representation of sequences in aptamer libraries and deep sequencing enables acyclic identification of aptamers. We demonstrated this by isolating a known family of aptamers for human alpha-thrombin. Aptamers were found within a library containing an average of 56,000 copies of each possible randomized 15mer segment. The high affinity sequences were counted many times above the background in 2-6 million reads. Clustering analysis of sequences with more than 10 counts distinguished two sequence motifs with candidates at high abundance. Motif I contained the previously observed consensus 15mer, Thb1 (46,000 counts), and related variants with mostly G/T substitutions; secondary analysis showed that affinity for thrombin correlated with abundance (K(d) = 12 nM for Thb1). The signal-to-noise ratio for this experiment was roughly 10,000?1 for Thb1. Motif II was unrelated to Thb1 with the leading candidate (29,000 counts) being a novel aptamer against hexose sugars in the storage and elution buffers for Concanavilin A (K(d) = 0.5 uM for alpha-methyl-mannoside); ConA was used to immobilize alpha-thrombin. CONCLUSIONS/SIGNIFICANCE: Over-representation together with deep sequencing can dramatically shorten the discovery process, distinguish aptamers having a wide range of affinity for the target, allow an exhaustive search of the sequence space within a simplified library, reduce the quantity of the target required, eliminate cycling artifacts, and should allow multiplexing of sequencing experiments and targets. PMID- 21625588 TI - Calpain-catalyzed proteolysis of human dUTPase specifically removes the nuclear localization signal peptide. AB - BACKGROUND: Calpain proteases drive intracellular signal transduction via specific proteolysis of multiple substrates upon Ca(2+)-induced activation. Recently, dUTPase, an enzyme essential to maintain genomic integrity, was identified as a physiological calpain substrate in Drosophila cells. Here we investigate the potential structural/functional significance of calpain-activated proteolysis of human dUTPase. METHODOLOGY/PRINCIPAL FINDINGS: Limited proteolysis of human dUTPase by mammalian m-calpain was investigated in the presence and absence of cognate ligands of either calpain or dUTPase. Significant proteolysis was observed only in the presence of Ca(II) ions, inducing calpain action. The presence or absence of the dUTP-analogue alpha,beta-imido-dUTP did not show any effect on Ca(2+)-calpain-induced cleavage of human dUTPase. The catalytic rate constant of dUTPase was unaffected by calpain cleavage. Gel electrophoretic analysis showed that Ca(2+)-calpain-induced cleavage of human dUTPase resulted in several distinctly observable dUTPase fragments. Mass spectrometric identification of the calpain-cleaved fragments identified three calpain cleavage sites (between residues (4)SE(5); (7)TP(8); and (31)LS(32)). The cleavage between the (31)LS(32) peptide bond specifically removes the flexible N-terminal nuclear localization signal, indispensable for cognate localization. CONCLUSIONS/SIGNIFICANCE: Results argue for a mechanism where Ca(2+)-calpain may regulate nuclear availability and degradation of dUTPase. PMID- 21625589 TI - Dual function of the NK cell receptor 2B4 (CD244) in the regulation of HCV specific CD8+ T cells. AB - The outcome of viral infections is dependent on the function of CD8+ T cells which are tightly regulated by costimulatory molecules. The NK cell receptor 2B4 (CD244) is a transmembrane protein belonging to the Ig superfamily which can also be expressed by CD8+ T cells. The aim of this study was to analyze the role of 2B4 as an additional costimulatory receptor regulating CD8+ T cell function and in particular to investigate its implication for exhaustion of hepatitis C virus (HCV)-specific CD8+ T cells during persistent infection. We demonstrate that (i) 2B4 is expressed on virus-specific CD8+ T cells during acute and chronic hepatitis C, (ii) that 2B4 cross-linking can lead to both inhibition and activation of HCV-specific CD8+ T cell function, depending on expression levels of 2B4 and the intracellular adaptor molecule SAP and (iii) that 2B4 stimulation may counteract enhanced proliferation of HCV-specific CD8+ T cells induced by PD1 blockade. We suggest that 2B4 is another important molecule within the network of costimulatory/inhibitory receptors regulating CD8+ T cell function in acute and chronic hepatitis C and that 2B4 expression levels could also be a marker of CD8+ T cell dysfunction. Understanding in more detail how 2B4 exerts its differential effects could have implications for the development of novel immunotherapies of HCV infection aiming to achieve immune control. PMID- 21625590 TI - Viral CTL escape mutants are generated in lymph nodes and subsequently become fixed in plasma and rectal mucosa during acute SIV infection of macaques. AB - SIV(mac239) infection of rhesus macaques (RMs) results in AIDS despite the generation of a strong antiviral cytotoxic T lymphocyte (CTL) response, possibly due to the emergence of viral escape mutants that prevent recognition of infected cells by CTLs. To determine the anatomic origin of these SIV mutants, we longitudinally assessed the presence of CTL escape variants in two MamuA*01 restricted immunodominant epitopes (Tat-SL8 and Gag-CM9) in the plasma, PBMCs, lymph nodes (LN), and rectal biopsies (RB) of fifteen SIV(mac239)-infected RMs. As expected, Gag-CM9 did not exhibit signs of escape before day 84 post infection. In contrast, Tat-SL8 escape mutants were apparent in all tissues by day 14 post infection. Interestingly LNs and plasma exhibited the highest level of escape at day 14 and day 28 post infection, respectively, with the rate of escape in the RB remaining lower throughout the acute infection. The possibility that CTL escape occurs in LNs before RBs is confirmed by the observation that the specific mutants found at high frequency in LNs at day 14 post infection became dominant at day 28 post infection in plasma, PBMC, and RB. Finally, the frequency of escape mutants in plasma at day 28 post infection correlated strongly with the level Tat-SL8-specific CD8 T cells in the LN and PBMC at day 14 post infection. These results indicate that LNs represent the primary source of CTL escape mutants during the acute phase of SIV(mac239) infection, suggesting that LNs are the main anatomic sites of virus replication and/or the tissues in which CTL pressure is most effective in selecting SIV escape variants. PMID- 21625591 TI - A gamma interferon independent mechanism of CD4 T cell mediated control of M. tuberculosis infection in vivo. AB - CD4 T cell deficiency or defective IFNgamma signaling render humans and mice highly susceptible to Mycobacterium tuberculosis (Mtb) infection. The prevailing model is that Th1 CD4 T cells produce IFNgamma to activate bactericidal effector mechanisms of infected macrophages. Here we test this model by directly interrogating the effector functions of Th1 CD4 T cells required to control Mtb in vivo. While Th1 CD4 T cells specific for the Mtb antigen ESAT-6 restrict in vivo Mtb growth, this inhibition is independent of IFNgamma or TNF and does not require the perforin or FAS effector pathways. Adoptive transfer of Th17 CD4 T cells specific for ESAT-6 partially inhibited Mtb growth while Th2 CD4 T cells were largely ineffective. These results imply a previously unrecognized IFNgamma/TNF independent pathway that efficiently controls Mtb and suggest that optimization of this alternative effector function may provide new therapeutic avenues to combat Mtb through vaccination. PMID- 21625592 TI - Vertical distribution of epibenthic freshwater cyanobacterial Synechococcus spp. strains depends on their ability for photoprotection. AB - BACKGROUND: Epibenthic cyanobacteria often grow in environments where the fluctuation of light intensity and quality is extreme and frequent. Different strategies have been developed to cope with this problem depending on the distribution of cyanobacteria in the water column. PRINCIPAL FINDINGS: Here we provide an experimental proof that the light intensity plays an important role in the vertical distribution of seven, closely related, epibenthic Synechococcus spp. strains isolated from various water depths from the littoral zone of Lake Constance in Germany and cultivated under laboratory conditions. Pigment analysis revealed that the amount of chlorophyll a and total carotenoids decreased with the time of light stress exposure in three phycoerythrin-rich strains collected from 7.0 m water depth and remained low during the recovery phase. In contrast, a constant level of chlorophyll a and either constant or enhanced levels of carotenoids were assayed in phycocyanin-rich strains collected from 1.0 and 0.5 m water depths. Protein analysis revealed that while the amount of biliproteins remained constant in all strains during light stress and recovery, the amount of D1 protein from photosystem II reaction centre was strongly reduced under light stress conditions in strains from 7.0 m and 1.0 m water depth, but not in strains collected from 0.5 m depth. CONCLUSION: Based on these data we propose that light intensity, in addition to light quality, is an important selective force in the vertical distribution of Synechococcus spp. strains, depending on their genetically fixed mechanisms for photoprotection. PMID- 21625593 TI - Discovery of nuclear-encoded genes for the neurotoxin saxitoxin in dinoflagellates. AB - Saxitoxin is a potent neurotoxin that occurs in aquatic environments worldwide. Ingestion of vector species can lead to paralytic shellfish poisoning, a severe human illness that may lead to paralysis and death. In freshwaters, the toxin is produced by prokaryotic cyanobacteria; in marine waters, it is associated with eukaryotic dinoflagellates. However, several studies suggest that saxitoxin is not produced by dinoflagellates themselves, but by co-cultured bacteria. Here, we show that genes required for saxitoxin synthesis are encoded in the nuclear genomes of dinoflagellates. We sequenced >1.2*10(6) mRNA transcripts from the two saxitoxin-producing dinoflagellate strains Alexandrium fundyense CCMP1719 and A. minutum CCMP113 using high-throughput sequencing technology. In addition, we used in silico transcriptome analyses, RACE, qPCR and conventional PCR coupled with Sanger sequencing. These approaches successfully identified genes required for saxitoxin-synthesis in the two transcriptomes. We focused on sxtA, the unique starting gene of saxitoxin synthesis, and show that the dinoflagellate transcripts of sxtA have the same domain structure as the cyanobacterial sxtA genes. But, in contrast to the bacterial homologs, the dinoflagellate transcripts are monocistronic, have a higher GC content, occur in multiple copies, contain typical dinoflagellate spliced-leader sequences and eukaryotic polyA-tails. Further, we investigated 28 saxitoxin-producing and non-producing dinoflagellate strains from six different genera for the presence of genomic sxtA homologs. Our results show very good agreement between the presence of sxtA and saxitoxin synthesis, except in three strains of A. tamarense, for which we amplified sxtA, but did not detect the toxin. Our work opens for possibilities to develop molecular tools to detect saxitoxin-producing dinoflagellates in the environment. PMID- 21625594 TI - P130Cas attenuates epidermal growth factor (EGF) receptor internalization by modulating EGF-triggered dynamin phosphorylation. AB - BACKGROUND: Endocytosis controls localization-specific signal transduction via epidermal growth factor receptor (EGFR), as well as downregulation of that receptor. Extracellular matrix (ECM)-integrin coupling induces formation of macromolecular complexes that include EGFR, integrin, Src kinase and p130Cas, resulting in EGFR activation. In addition, cell adhesion to ECM increases EGFR localization at the cell surface and reduces EGFR internalization. The molecular mechanisms involved are not yet well understood. METHODOLOGY/PRINCIPAL FINDINGS: We investigated the molecular mechanism by which p130Cas affects the endocytic regulation of EGFR. Biochemical quantification revealed that cell adhesion to fibronectin (FN) increases total EGFR levels and its phosphorylation, and that p130Cas is required for this process. Measurements of Texas Red-labeled EGF uptake and cell surface EGFR revealed that p130Cas overexpression reduces EGF induced EGFR internalization, while p130Cas depletion enhances it. In addition, both FN-mediated cell adhesion and p130Cas overexpression reduce EGF-stimulated dynamin phosphorylation, which is necessary for EGF-induced EGFR internalization. Coimmunoprecipitation and GST pull-down assays confirmed the interaction between p130Cas and dynamin. Moreover, a SH3-domain-deleted form of p130Cas, which shows diminished binding to dynamin, inhibits dynamin phosphorylation and EGF uptake less effectively than wild-type p130Cas. CONCLUSIONS/SIGNIFICANCE: Our results show that p130Cas plays an inhibitory role in EGFR internalization via its interaction with dynamin. Given that the EGFR internalization process determines signaling density and specificity in the EGFR pathway, these findings suggest that the interaction between p130Cas and dynamin may regulate EGFR trafficking and signaling in the same manner as other endocytic regulatory proteins related to EGFR endocytosis. PMID- 21625595 TI - Phage encoded H-NS: a potential achilles heel in the bacterial defence system. AB - The relationship between phage and their microbial hosts is difficult to elucidate in complex natural ecosystems. Engineered systems performing enhanced biological phosphorus removal (EBPR), offer stable, lower complexity communities for studying phage-host interactions. Here, metagenomic data from an EBPR reactor dominated by Candidatus Accumulibacter phosphatis (CAP), led to the recovery of three complete and six partial phage genomes. Heat-stable nucleoid structuring (H NS) protein, a global transcriptional repressor in bacteria, was identified in one of the complete phage genomes (EPV1), and was most similar to a homolog in CAP. We infer that EPV1 is a CAP-specific phage and has the potential to repress up to 6% of host genes based on the presence of putative H-NS binding sites in the CAP genome. These genes include CRISPR associated proteins and a Type III restriction-modification system, which are key host defense mechanisms against phage infection. Further, EPV1 was the only member of the phage community found in an EBPR microbial metagenome collected seven months prior. We propose that EPV1 laterally acquired H-NS from CAP providing it with a means to reduce bacterial defenses, a selective advantage over other phage in the EBPR system. Phage encoded H-NS could constitute a previously unrecognized weapon in the phage host arms race. PMID- 21625596 TI - Clinical value of prognostic instruments to identify patients with an increased risk for osteoporotic fractures: systematic review. AB - BACKGROUND: With the broad availability of effective medications, identifying individuals bearing a higher risk for osteoporotic fractures has become an issue of major concern in modern medicine. In recent years various prognostic instruments have become available showing conflicting results regarding estimated risks for individual patients. OBJECTIVE: To provide an overview of current evidence and of opportunities for further research. METHODOLOGY/PRINCIPAL FINDINGS: Systematic review: We identified studies describing the development of instruments and all subsequent validations in electronic databases and reference lists of included studies. We screened for inclusion, read full papers and extracted data on salient clinical features, performance characteristics and quality in duplicate. Searches retrieved 5,275 records of which full texts of 167 papers were obtained after screening titles and abstract. We included 35 studies enrolling a total of 609,969 patients (median 2546) reporting on 31 derivations and 12 validations after assessing full texts. Median follow-up time was 4.1 years (IQR 3 to 7.7). Only four studies validated an instrument that was developed by another group. None of the existing instruments was validated more than once. The five most frequent included variables in the final model were age, body mass index, bone mass index, past history of falls, and maternal history of fractures. The methodological quality of the studies was moderate. CONCLUSION: There is a plethora of evidence available studying the association of risk profiles and the development of osteoporotic fractures. The small number of out of-sample validations, the large variety of study characteristics, outcomes and follow-up periods impedes from deriving robust summaries and from conclusions regarding the clinical performance of many tools. First and foremost, future activity in this field should aim at reaching a consensus among clinical experts in respect to the existing instruments. Then we call for careful validations and expedient adaptations for local circumstances of the most promising candidates. PMID- 21625597 TI - Two naturally occurring terpenes, dehydrocostuslactone and costunolide, decrease intracellular GSH content and inhibit STAT3 activation. AB - The main purpose of the present study is to envisage the molecular mechanism of inhibitory action of dehydrocostuslactone (DCE) and costunolide (CS), two naturally occurring sesquiterpene lactones, towards the activation of signal transducer and activator of transcription 3 (STAT3). We report that, in human THP 1 cell line, they inhibit IL-6-elicited tyrosine phosphorylation of STAT3 and its DNA binding activity with EC(50) of 10 uM with concomitant down-regulation of the phosphorylation of the tyrosine Janus kinases JAK1, JAK2 and Tyk2. Furthermore, these compounds that contain an alpha-beta-unsaturated carbonyl moiety and function as potent Michael reaction acceptor, induce a rapid drop in intracellular glutathione (GSH) concentration by direct interaction with it, thereby triggering S-glutathionylation of STAT3. Dehydrocostunolide (HCS), the reduced form of CS lacking only the alpha-beta-unsaturated carbonyl group, fails to exert any inhibitory action. Finally, the glutathione ethylene ester (GEE), the cell permeable GSH form, reverts the inhibitory action of DCE and CS on STAT3 tyrosine phosphorylation. We conclude that these two sesquiterpene lactones are able to induce redox-dependent post-translational modification of cysteine residues of STAT3 protein in order to regulate its function. PMID- 21625598 TI - Cell-intrinsic NF-kappaB activation is critical for the development of natural regulatory T cells in mice. AB - BACKGROUND: Naturally occurring CD4(+)CD25(+)Foxp3(+) regulatory T (Treg) cells develop in the thymus and represent a mature T cell subpopulation critically involved in maintaining peripheral tolerance. The differentiation of Treg cells in the thymus requires T cell receptor (TCR)/CD28 stimulation along with cytokine promoted Foxp3 induction. TCR-mediated nuclear factor kappa B (NF-kappaB) activation seems to be involved in differentiation of Treg cells because deletion of components of the NF-kappaB signaling pathway, as well as of NF-kappaB transcription factors, leads to markedly decreased Treg cell numbers in thymus and periphery. METHODOLOGY/PRINCIPAL FINDINGS: To investigate if Treg cell intrinsic NF-kappaB activation is required for thymic development and peripheral homeostasis of Treg cells we used transgenic (Tg) mice with thymocyte-specific expression of a stable IkappaBalpha mutant to inhibit NF-kappaB activation solely within the T cell lineage. Here we show that Treg cell-intrinsic NF-kappaB activation is important for the generation of cytokine-responsive Foxp3(-) thymic Treg precursors and their further differentiation into mature Treg cells. Treg cell development could neither be completely rescued by the addition of exogenous Interleukin 2 (IL-2) nor by the presence of wild-type derived cells in adoptive transfer experiments. However, peripheral NF-kappaB activation appears to be required for IL-2 production by conventional T cells, thereby participating in Treg cell homeostasis. Moreover, pharmacological NF-kappaB inhibition via the IkappaB kinase beta (IKKbeta) inhibitor AS602868 led to markedly diminished thymic and peripheral Treg cell frequencies. CONCLUSION/SIGNIFICANCE: Our results indicate that Treg cell-intrinsic NF-kappaB activation is essential for thymic Treg cell differentiation, and further suggest pharmacological NF-kappaB inhibition as a potential therapeutic approach for manipulating this process. PMID- 21625599 TI - C-terminus glycans with critical functional role in the maturation of secretory glycoproteins. AB - The N-glycans of membrane glycoproteins are mainly exposed to the extracellular space. Human tyrosinase is a transmembrane glycoprotein with six or seven bulky N glycans exposed towards the lumen of subcellular organelles. The central active site region of human tyrosinase is modeled here within less than 2.5 A accuracy starting from Streptomyces castaneoglobisporus tyrosinase. The model accounts for the last five C-terminus glycosylation sites of which four are occupied and indicates that these cluster in two pairs--one in close vicinity to the active site and the other on the opposite side. We have analyzed and compared the roles of all tyrosinase N-glycans during tyrosinase processing with a special focus on the proximal to the active site N-glycans, s6:N337 and s7:N371, versus s3:N161 and s4:N230 which decorate the opposite side of the domain. To this end, we have constructed mutants of human tyrosinase in which its seven N-glycosylation sites were deleted. Ablation of the s6:N337 and s7:N371 sites arrests the post translational productive folding process resulting in terminally misfolded mutants subjected to degradation through the mannosidase driven ERAD pathway. In contrast, single mutants of the other five N-glycans located either opposite to the active site or into the N-terminus Cys1 extension of tyrosinase are temperature-sensitive mutants and recover enzymatic activity at the permissive temperature of 31 degrees C. Sites s3 and s4 display selective calreticulin binding properties. The C-terminus sites s7 and s6 are critical for the endoplasmic reticulum retention and intracellular disposal. Results herein suggest that individual N-glycan location is critical for the stability, regional folding control and secretion of human tyrosinase and explains some tyrosinase gene missense mutations associated with oculocutaneous albinism type I. PMID- 21625600 TI - Association of HbA1c values with mortality and cardiovascular events in diabetic dialysis patients. The INVOR study and review of the literature. AB - BACKGROUND: Improved glycemic control reduces complications in patients with diabetes mellitus (DM). However, it is discussed controversially whether patients with diabetes mellitus and end-stage renal disease benefit from strict glycemic control. METHODS: We followed 78 patients with DM initiating dialysis treatment of the region of Vorarlberg in a prospective cohort study applying a time dependent Cox regression analysis using all measured laboratory values for up to more than seven years. This resulted in 880 HbA(1c) measurements (with one measurement every 3.16 patient months on average) during the entire observation period. Non-linear P-splines were used to allow flexible modeling of the association with mortality and cardiovascular disease (CVD) events. RESULTS: We observed a decreased mortality risk with increasing HbA(1c) values (HR = 0.72 per 1% increase, p = 0.024). Adjustment for age and sex and additional adjustment for other CVD risk factors only slightly attenuated the association (HR = 0.71, p = 0.044). A non-linear P-spline showed that the association did not follow a fully linear pattern with a highly significant non-linear component (p = 0.001) with an increased risk of all-cause mortality for HbA(1c) values up to 6-7%. Causes of death were associated with HbA(1c) values. The risk for CVD events, however, increased with increasing HbA(1c) values (HR = 1.24 per 1% increase, p = 0.048) but vanished after extended adjustments. CONCLUSIONS: This study considered the entire information collected on HbA(1c) over a period of more than seven years. Besides the methodological advantages our data indicate a significant inverse association between HbA(1c) levels and all-cause mortality. However, for CVD events no significant association could be found. PMID- 21625601 TI - Exploiting nucleotide composition to engineer promoters. AB - The choice of promoter is a critical step in optimizing the efficiency and stability of recombinant protein production in mammalian cell lines. Artificial promoters that provide stable expression across cell lines and can be designed to the desired strength constitute an alternative to the use of viral promoters. Here, we show how the nucleotide characteristics of highly active human promoters can be modelled via the genome-wide frequency distribution of short motifs: by overlapping motifs that occur infrequently in the genome, we constructed contiguous sequence that is rich in GC and CpGs, both features of known promoters, but lacking homology to real promoters. We show that snippets from this sequence, at 100 base pairs or longer, drive gene expression in vitro in a number of mammalian cells, and are thus candidates for use in protein production. We further show that expression is driven by the general transcription factors TFIIB and TFIID, both being ubiquitously present across cell types, which results in less tissue- and species-specific regulation compared to the viral promoter SV40. We lastly found that the strength of a promoter can be tuned up and down by modulating the counts of GC and CpGs in localized regions. These results constitute a "proof-of-concept" for custom-designing promoters that are suitable for biotechnological and medical applications. PMID- 21625602 TI - The Arabidopsis thaliana homeobox gene ATHB12 is involved in symptom development caused by geminivirus infection. AB - BACKGROUND: Geminiviruses are single-stranded DNA viruses that infect a number of monocotyledonous and dicotyledonous plants. Arabidopsis is susceptible to infection with the Curtovirus, Beet severe curly top virus (BSCTV). Infection of Arabidopsis with BSCTV causes severe symptoms characterized by stunting, leaf curling, and the development of abnormal inflorescence and root structures. BSCTV induced symptom development requires the virus-encoded C4 protein which is thought to interact with specific plant-host proteins and disrupt signaling pathways important for controlling cell division and development. Very little is known about the specific plant regulatory factors that participate in BSCTV induced symptom development. This study was conducted to identify specific transcription factors that are induced by BSCTV infection. METHODOLOGY/PRINCIPAL FINDINGS: Arabidopsis plants were inoculated with BSCTV and the induction of specific transcription factors was monitored using quantitative real-time polymerase chain reaction assays. We found that the ATHB12 and ATHB7 genes, members of the homeodomain-leucine zipper family of transcription factors previously shown to be induced by abscisic acid and water stress, are induced in symptomatic tissues of Arabidopsis inoculated with BSCTV. ATHB12 expression is correlated with an array of morphological abnormalities including leaf curling, stunting, and callus-like structures in infected Arabidopsis. Inoculation of plants with a BSCTV mutant with a defective c4 gene failed to induce ATHB12. Transgenic plants expressing the BSCTV C4 gene exhibited increased ATHB12 expression whereas BSCTV-infected ATHB12 knock-down plants developed milder symptoms and had lower ATHB12 expression compared to the wild-type plants. Reporter gene studies demonstrated that the ATHB12 promoter was responsive to BSCTV infection and the highest expression levels were observed in symptomatic tissues where cell cycle genes also were induced. CONCLUSIONS/SIGNIFICANCE: These results suggest that ATHB7 and ATHB12 may play an important role in the activation of the abnormal cell division associated with symptom development during geminivirus infection. PMID- 21625603 TI - Are algae relevant to the detritus-based food web in tank-bromeliads? AB - We assessed the occurrence of algae in five species of tank-bromeliads found in contrasting environmental sites in a Neotropical, primary rainforest around the Nouragues Research Station, French Guiana. The distributions of both algal abundance and biomass were examined based on physical parameters, the morphological characteristics of bromeliad species and with regard to the structure of other aquatic microbial communities held in the tanks. Algae were retrieved in all of the bromeliad species with mean densities ranging from ~10(2) to 10(4) cells/mL. Their biomass was positively correlated to light exposure and bacterial biomass. Algae represented a tiny component of the detrital food web in shaded bromeliads but accounted for up to 30 percent of the living microbial carbon in the tanks of Catopsis berteroniana, located in a highly exposed area. Thus, while nutrient supplies are believed to originate from wind-borne particles and trapped insects (i.e., allochtonous organic matter), our results indicate that primary producers (i.e., autochtonous organic matter) are present in this insectivorous bromeliad. Using a 24-h incubation of size-fractionated and manipulated samples from this plant, we evaluated the impact of mosquito foraging on algae, other microorganisms and rotifers. The prey assemblages were greatly altered by the predation of mosquito larvae. Grazing losses indicated that the dominant algal taxon, Bumilleriopsis sp., like protozoa and rotifers, is a significant part of the diet of mosquito larvae. We conclude that algae are a relevant functional community of the aquatic food web in C. berteroniana and might form the basis of a complementary non-detrital food web. PMID- 21625604 TI - Advanced autumn migration of sparrowhawk has increased the predation risk of long distance migrants in Finland. AB - Predation affects life history traits of nearly all organisms and the population consequences of predator avoidance are often larger than predation itself. Climate change has been shown to cause phenological changes. These changes are not necessarily similar between species and may cause mismatches between prey and predator. Eurasian sparrowhawk Accipiter nisus, the main predator of passerines, has advanced its autumn phenology by about ten days in 30 years due to climate change. However, we do not know if sparrowhawk migrate earlier in response to earlier migration by its prey or if earlier sparrowhawk migration results in changes to predation risk on its prey. By using the median departure date of 41 passerine species I was able to show that early migrating passerines tend to advance, and late migrating species delay their departure, but none of the species have advanced their departure times as much as the sparrowhawk. This has lead to a situation of increased predation risk on early migrating long-distance migrants (LDM) and decreased the overlap of migration season with later departing short-distance migrants (SDM). Findings highlight the growing list of problems of declining LDM populations caused by climate change. On the other hand it seems that the autumn migration may become safer for SDM whose populations are growing. Results demonstrate that passerines show very conservative response in autumn phenology to climate change, and thus phenological mismatches caused by global warming are not necessarily increasing towards the higher trophic levels. PMID- 21625605 TI - Human glycolipid transfer protein (GLTP) expression modulates cell shape. AB - Glycolipid transfer protein (GLTP) accelerates glycosphingolipid (GSL) intermembrane transfer via a unique lipid transfer/binding fold (GLTP-fold) that defines the GLTP superfamily and is the prototype for GLTP-like domains in larger proteins, i.e. phosphoinositol 4-phosphate adaptor protein-2 (FAPP2). Although GLTP-folds are known to play roles in the nonvesicular intracellular trafficking of glycolipids, their ability to alter cell phenotype remains unexplored. In the present study, overexpression of human glycolipid transfer protein (GLTP) was found to dramatically alter cell phenotype, with cells becoming round between 24 and 48 h after transfection. By 48 h post transfection, ~70% conversion to the markedly round shape was evident in HeLa and HEK-293 cells, but not in A549 cells. In contrast, overexpression of W96A-GLTP, a liganding-site point mutant with abrogated ability to transfer glycolipid, did not alter cell shape. The round adherent cells exhibited diminished motility in wound healing assays and an inability to endocytose cholera toxin but remained viable and showed little increase in apoptosis as assessed by poly(ADP-ribose) polymerase cleavage. A round cell phenotype also was induced by overexpression of FAPP2, which binds/transfers glycolipid via its C-terminal GLTP-like fold, but not by a plant GLTP ortholog (ACD11), which is incapable of glycolipid binding/transfer. Screening for human protein partners of GLTP by yeast two hybrid screening and by immuno-pulldown analyses revealed regulation of the GLTP-induced cell rounding response by interaction with delta-catenin. Remarkably, while delta-catenin overexpression alone induced dendritic outgrowths, coexpression of GLTP along with delta-catenin accelerated transition to the rounded phenotype. The findings represent the first known phenotypic changes triggered by GLTP overexpression and regulated by direct interaction with a p120-catenin protein family member. PMID- 21625606 TI - Comparative transcriptome analysis of Bacillus subtilis responding to dissolved oxygen in adenosine fermentation. AB - Dissolved oxygen (DO) is an important factor for adenosine fermentation. Our previous experiments have shown that low oxygen supply in the growth period was optimal for high adenosine yield. Herein, to better understand the link between oxygen supply and adenosine productivity in B. subtilis (ATCC21616), we sought to systematically explore the effect of DO on genetic regulation and metabolism through transcriptome analysis. The microarrays representing 4,106 genes were used to study temporal transcript profiles of B. subtilis fermentation in response to high oxygen supply (agitation 700 r/min) and low oxygen supply (agitation 450 r/min). The transcriptome data analysis revealed that low oxygen supply has three major effects on metabolism: enhance carbon metabolism (glucose metabolism, pyruvate metabolism and carbon overflow), inhibit degradation of nitrogen sources (glutamate family amino acids and xanthine) and purine synthesis. Inhibition of xanthine degradation was the reason that low oxygen supply enhanced adenosine production. These provide us with potential targets, which can be modified to achieve higher adenosine yield. Expression of genes involved in energy, cell type differentiation, protein synthesis was also influenced by oxygen supply. These results provided new insights into the relationship between oxygen supply and metabolism. PMID- 21625607 TI - Identification of Mycobacterium tuberculosis-specific Th1, Th17 and Th22 cells using the expression of CD40L in tuberculous pleurisy. AB - Important advances have been made in the immunodiagnosis of tuberculosis (TB) based on the detection of Mycobacterium tuberculosis (MTB)-specific T cells. However, the sensitivity and specificity of the immunological approach are relatively low because there are no specific markers for antigen-specific Th cells, and some of the Th cells that do not produce cytokines can be overlooked using this approach. In this study, we found that MTB-specific peptides of ESAT 6/CFP-10 can stimulate the expression of CD40L specifically in CD4(+) T cells but not other cells from pleural fluid cells (PFCs) in patients with tuberculous pleurisy (TBP). CD4(+)CD40L(+) but not CD4(+)CD40L(-) T cells express IFN-gamma, IL-2, TNF-alpha, IL-17 or IL-22 after stimulation with MTB-specific peptides. In addition, CD4(+)CD40L(+) T cells were found to be mostly polyfunctional T cells that simultaneously produce IFN-gamma, IL-2 and TNF-alpha and display an effector or effector memory phenotype (CD45RA(-)CD45RO(+)CCR7(-)CD62L(-)ICOS(-)). To determine the specificity of CD4(+)CD40L(+) T cells, we incubated PFCs with ESTA 6/CFP-10 peptides and sorted live CD4(+)CD40L(+) and CD4(+)CD40L(-) T cells by flow cytometry. We further demonstrated that sorted CD4(+)CD40L(+), but not CD4(+)CD40L(-) fractions, principally produced IFN-gamma, IL-2, TNF-alpha, IL-17 and IL-22 following restimulation with ESTA-6/CFP-10 peptides. Taken together, our data indicate that the expression of CD40L on MTB-specific CD4(+) T cells could be a good marker for the evaluation and isolation of MTB-specific Th cells and might also be useful in the diagnosis of TB. PMID- 21625608 TI - Dendritic cells exposed to MVA-based HIV-1 vaccine induce highly functional HIV-1 specific CD8(+) T cell responses in HIV-1-infected individuals. AB - Currently, MVA virus vectors carrying HIV-1 genes are being developed as HIV 1/AIDS prophylactic/therapeutic vaccines. Nevertheless, little is known about the impact of these vectors on human dendritic cells (DC) and their capacity to present HIV-1 antigens to human HIV-specific T cells. This study aimed to characterize the interaction of MVA and MVA expressing the HIV-1 genes Env-Gag Pol-Nef of clade B (referred to as MVA-B) in human monocyte-derived dendritic cells (MDDC) and the subsequent processes of HIV-1 antigen presentation and activation of memory HIV-1-specific T lymphocytes. For these purposes, we performed ex vivo assays with MDDC and autologous lymphocytes from asymptomatic HIV-infected patients. Infection of MDDC with MVA-B or MVA, at the optimal dose of 0.3 PFU/MDDC, induced by itself a moderate degree of maturation of MDDC, involving secretion of cytokines and chemokines (IL1-ra, IL-7, TNF-alpha, IL-6, IL-12, IL-15, IL-8, MCP-1, MIP-1alpha, MIP-1beta, RANTES, IP-10, MIG, and IFN alpha). MDDC infected with MVA or MVA-B and following a period of 48 h or 72 h of maturation were able to migrate toward CCL19 or CCL21 chemokine gradients. MVA-B infection induced apoptosis of the infected cells and the resulting apoptotic bodies were engulfed by the uninfected MDDC, which cross-presented HIV-1 antigens to autologous CD8(+) T lymphocytes. MVA-B-infected MDDC co-cultured with autologous T lymphocytes induced a highly functional HIV-specific CD8(+) T cell response including proliferation, secretion of IFN-gamma, IL-2, TNF-alpha, MIP 1beta, MIP-1alpha, RANTES and IL-6, and strong cytotoxic activity against autologous HIV-1-infected CD4(+) T lymphocytes. These results evidence the adjuvant role of the vector itself (MVA) and support the clinical development of prophylactic and therapeutic anti-HIV vaccines based on MVA-B. PMID- 21625609 TI - An exploration of the control of micturition using a novel in situ arterially perfused rat preparation. AB - Our goal was to develop and refine a decerebrate arterially perfused rat (DAPR) preparation that allows the complete bladder filling and voiding cycle to be investigated without some of the restrictions inherent with in vivo experimentation [e.g., ease and speed of set up (30 min), control over the extracellular milieu and free of anesthetic agents]. Both spontaneous (naturalistic bladder filling from ureters) and evoked (in response to intravesical infusion) voids were routinely and reproducibly observed which had similar pressure characteristics. The DAPR allows the simultaneous measurement of bladder intra-luminal pressure, external urinary sphincter-electromyogram (EUS EMG), pelvic afferent nerve activity, pudendal motor activity, and permits excellent visualization of the entire lower urinary tract, during typical rat filling and voiding responses. The voiding responses were modulated or eliminated by interventions at a number of levels including at the afferent terminal fields (intravesical capsaicin sensitization-desensitization), autonomic (ganglion blockade with hexamethonium), and somatic motor (vecuronium block of the EUS) outflow and required intact brainstem/hindbrain-spinal coordination (as demonstrated by sequential hindbrain transections). Both innocuous (e.g., perineal stimulation) and nociceptive (tail/paw pinch) somatic stimuli elicited an increase in EUS-EMG indicating intact sensory feedback loops. Spontaneous non micturition contractions were observed between fluid infusions at a frequency and amplitude of 1.4 +/- 0.9 per minute and 1.4 +/- 0.3 mmHg, respectively and their amplitude increased when autonomic control was compromised. In conclusion, the DAPR is a tractable and useful model for the study of neural bladder control showing intact afferent signaling, spinal and hindbrain co-ordination and efferent control over the lower urinary tract end organs and can be extended to study bladder pathologies and trial novel treatments. PMID- 21625610 TI - Computational detection of alternative exon usage. AB - BACKGROUND: With the advent of the GeneChip Exon Arrays, it is now possible to extract "exon-level" expression estimates, allowing for detection of alternative splicing events, one of the primary mechanisms of transcript diversity. In the context of (1) a complex trait use case and (2) a human cerebellum vs. heart comparison on previously validated data, we present a transcript-based statistical model and validation framework to allow detection of alternative exon usage (AEU) between different groups. To illustrate the approach, we detect and confirm differences in exon usage in the two of the most widely studied mouse genetic models (the C57BL/6J and DBA/2J inbred strains) and in a human dataset. RESULTS: We developed a computational framework that consists of probe level annotation mapping and statistical modeling to detect putative AEU events, as well as visualization and alignment with known splice events. We show a dramatic improvement (~25 fold) in the ability to detect these events using the appropriate annotation and statistical model which is actually specified at the transcript level, as compared with the transcript cluster/gene-level annotation used on the array. An additional component of this workflow is a probe index that allows ranking AEU candidates for validation and can aid in identification of false positives due to single nucleotide polymorphisms. DISCUSSION: Our work highlights the importance of concordance between the functional unit interrogated (e.g., gene, transcripts) and the entity (e.g., exon, probeset) within the statistical model. The framework we present is broadly applicable to other platforms (including RNAseq). PMID- 21625611 TI - Cortical potentials evoked by deep brain stimulation in the subthalamic area. AB - Deep brain stimulation (DBS) of the subthalamic nucleus (STN) has been used since the mid-1990s as a treatment for patients with Parkinson's disease, and more recently also in other conditions, such as dystonia or obsessive compulsive disorder. Non-invasive studies of cortical evoked potentials (EPs) that follow individual STN-DBS stimuli has provided us with insights about the conduction of the DBS pulses to the cortex. Such EPs have multiple components of different latencies, making it possible to distinguish short-latency and long-latency responses (3-8 ms and 18-25 ms latency, respectively). The available evidence indicates that these short- and long-latency EPs correspond to conduction from the STN stimulation site to the cortical recording location via anti- and orthodromic pathways, respectively. In this review we survey the literature from recording studies in human patients treated with STN-DBS for Parkinson's disease and other conditions, as well as recent animal studies (including our own) that have begun to elucidate details of the pathways, frequency dependencies, and other features of EPs. In addition, we comment on the possible clinical utility of this knowledge. PMID- 21625612 TI - Combining confocal laser scanning microscopy with serial section reconstruction in the study of adult neurogenesis. AB - Current advances in imaging techniques have extended the possibility of visualizing small structures within large volumes of both fixed and live specimens without sectioning. These techniques have contributed valuable information to study neuronal plasticity in the adult brain. However, technical limits still hamper the use of these approaches to investigate neurogenic regions located far from the ventricular surface such as parenchymal neurogenic niches, or the scattered neuroblasts induced by brain lesions. Here, we present a method to combine confocal laser scanning microscopy (CLSM) and serial section reconstruction in order to reconstruct large volumes of brain tissue at cellular resolution. In this method a series of thick sections are imaged with CLSM and the resulting stacks of images are registered and 3D reconstructed. This approach is based on existing freeware software and can be performed on ordinary laboratory personal computers. By using this technique we have investigated the morphology and spatial organization of a group of doublecortin (DCX)+ neuroblasts located in the lateral striatum of the late post-natal guinea pig. The 3D study unraveled a complex network of long and poorly ramified cell processes, often fascicled and mostly oriented along the internal capsule fiber bundles. These data support CLSM serial section reconstruction as a reliable alternative to the whole mount approaches to analyze cyto-architectural features of adult germinative niches. PMID- 21625613 TI - Necrotizing fasciitis versus pyoderma gangrenosum: securing the correct diagnosis! A case report and literature review. AB - OBJECTIVE: To highlight the key differences in history, examination, and management of pyoderma gangrenosum and necrotizing fasciitis and to outline the importance of distinguishing these 2 conditions. METHOD: We present a case report of a gentleman with a background of ulcerative colitis having a 1-week history of an erythematous wound and localized abscess to the right leg that failed to respond to antibiotic treatment and later on to surgical debridement of a presumed necrotizing fasciitis. Following referral to our plastic surgery unit, a diagnosis of pyoderma gangrenosum was made and this was confirmed following a response to steroid therapy within 48 hours. A literature review of pyoderma gangrenosum cases misdiagnosed for necrotizing fasciitis was carried out to compare and contrast pitfalls in misdiagnosing these 2 conditions. RESULTS: Literature review of 10 cases confirmed the association of pyoderma gangrenosum with inflammatory bowel disease, hematological disease, and surgical trauma. The presence of necrotic tissue in a pyoderma gangrenosum lesion can be a diagnostic pitfall; although blood and tissue culture investigations are usually negative in pyoderma gangrenosum, this may not always be the case. Inflammatory markers can be significantly high in pyoderma gangrenosum and pyrexia is not a feature limited to necrotizing fasciitis. CONCLUSIONS: Inappropriate surgical debridement of pyoderma gangrenosum can cause rapid extension of the lesion by enhancing the posttraumatic response and lead to potential reconstructive challenges with psychological repercussions. On the contrary, treating necrotizing fasciitis with immunosuppressive therapy may worsen the condition. The importance of understanding the pathogenesis, clinical features, and management of both conditions cannot be overemphasized. PMID- 21625614 TI - Upper lip Basal cell carcinoma reconstruction based on anatomical characteristics using skin-mucosa double opposing v-y advancement flap. AB - BACKGROUND: Upper lip defects less than one-fourth of total upper lip length are typically closed directly, but larger defects require reconstruction. Established techniques, such as the Abbe/Estlander flap, often require multiple procedures. This report discusses a single-step method, involving a skin-mucosa double opposing V-Y advancement flap, which was utilized to reconstruct the upper lip after basal cell carcinoma extirpation. METHODS: The patient is a 54-year-old woman who presented with a 2.5-cm basal cell carcinoma in the right upper lip. Two parallel V-Y flaps were designed, on "parallel" planes of the skin and mucosa, and were advanced following wedge resection of tumor and portion of the adjacent orbicularis oris muscle. Skin/mucosa flap edges were brought to create a new skin-vermilion border, which was then sutured onto existing skin-vermilion border edges. RESULTS: The patient was evaluated 6 months after the operation, and examination showed well-healed flaps with excellent aesthetic and functional results. There was a thin, marginal scar, where reconstructed vermilion (medial) edge was sutured to native vermilion. CONCLUSIONS: It was considered that this method could reconstruct anatomical features and function of the lip precisely. It seems that within certain limits (probably between one-fourth and one-third of total upper lip length), skin-mucosa double opposing V-Y advancement flaps appear to be the preferred method for upper lip reconstruction. PMID- 21625615 TI - CD38 exacerbates focal cytokine production, postischemic inflammation and brain injury after focal cerebral ischemia. AB - BACKGROUND: Converging evidence suggests that inflammatory processes significantly influence brain injury and clinical impairment in ischemic stroke. Although early studies suggested a key role of lymphocytes, recent data has emphasized the orchestrating function of innate immunity, i.e., macrophages and microglia. The bifunctional receptor and ectoenzyme CD38 synthesizes calcium mobilizing second messengers (e.g., cyclic ADP-ribose), which have been shown to be necessary for activation and migration of myeloid immune cells. Therefore, we investigated the dynamics of CD38 in stroke and the impact of CD38-deficiency on cytokine production, inflammation and cerebral damage in a mouse model of cerebral ischemia-reperfusion. METHODOLOGY/PRINCIPAL FINDINGS: We show that the local expression of the chemokine MCP-1 was attenuated in CD38-deficient mice compared with wildtype mice after focal cerebral ischemia and reperfusion. In contrast, no significant induction of MCP-1 expression was observed in peripheral blood after 6 hours. Flow cytometry analysis revealed less infiltrating macrophages and lymphocytes in the ischemic hemisphere of CD38-deficient mice, whereas the amount of resident microglia was unaltered. An up-regulation of CD38 expression was observed in macrophages and CD8(+) cells after focal cerebral ischemia in wildtype mice, whereas CD38 expression was unchanged in microglia. Finally, we demonstrate that CD38-deficiency decreases the cerebral ischemic injury and the persistent neurological deficit after three days of reperfusion in this murine temporary middle cerebral artery occlusion (tMCAO) model. CONCLUSION/SIGNIFICANCE: CD38 is differentially regulated following stroke and its deficiency attenuates the postischemic chemokine production, the immune cell infiltration and the cerebral injury after temporary ischemia and reperfusion. Therefore CD38 might prove a therapeutic target in ischemic stroke. PMID- 21625616 TI - Depilatory wax burns: experience and investigation. AB - OBJECTIVES: To retrospectively collect data on patients with burn injury due to hot depilatory wax. To investigate the effect of varying microwave output power on wax temperature. To determine whether instructions provided by manufacturers allow safe domestic use. METHODS: Data from the RAH burns database was collected for patients with wax-induced burns between January 1991 and January 2010. Wax temperatures were tested in a pilot study (4 wax products heated in microwave with power outputs of 800 W, 900 W, and 1100 (W) and a definitive study (5 wax products, 3 of each, heated in microwave with power outputs of 800 W, 1000 W, and 1200 (W). A number of different heating regimens were employed and temperatures were recorded using an infrared thermometer. RESULTS: Twenty-one patients were studied. Mean age was 26.5 years. The majority of burns were superficial (33.3%) or partial thickness (25.8%). The right hand was most commonly affected (38.1%), the mean total body surface area was 1%. The pilot study revealed an increase in wax temperature with the number of times the wax was heated. During definitive wax temperature testing, the maximum wax temperature recorded was 108.5 degrees C. Seventeen of 60 wax surface temperatures recorded exceeded 90 degrees C, 9 exceeded 100 degrees C. Ninety-three percent of the stirred wax temperatures showed an increase in wax temperature with an increase in microwave power output. CONCLUSION: Microwave-heated hair-removal wax has the potential to reach unsafe temperatures and cause burn injury, even when manufacture's heating instructions are followed. Safe use in domestic setting requires improvements in instructions provided by the manufacturer. PMID- 21625617 TI - Inactivation of chk2 and mus81 leads to impaired lymphocytes development, reduced genomic instability, and suppression of cancer. AB - Chk2 is an effector kinase important for the activation of cell cycle checkpoints, p53, and apoptosis in response to DNA damage. Mus81 is required for the restart of stalled replication forks and for genomic integrity. Mus81(Deltaex3-4/Deltaex3-4) mice have increased cancer susceptibility that is exacerbated by p53 inactivation. In this study, we demonstrate that Chk2 inactivation impairs the development of Mus81(Deltaex3-4/Deltaex3-4) lymphoid cells in a cell-autonomous manner. Importantly, in contrast to its predicted tumor suppressor function, loss of Chk2 promotes mitotic catastrophe and cell death, and it results in suppressed oncogenic transformation and tumor development in Mus81(Deltaex3-4/Deltaex3-4) background. Thus, our data indicate that an important role for Chk2 is maintaining lymphocyte development and that dual inactivation of Chk2 and Mus81 remarkably inhibits cancer. PMID- 21625618 TI - Prion formation and polyglutamine aggregation are controlled by two classes of genes. AB - Prions are self-perpetuating aggregated proteins that are not limited to mammalian systems but also exist in lower eukaryotes including yeast. While much work has focused around chaperones involved in prion maintenance, including Hsp104, little is known about factors involved in the appearance of prions. De novo appearance of the [PSI+] prion, which is the aggregated form of the Sup35 protein, is dramatically enhanced by transient overexpression of SUP35 in the presence of the prion form of the Rnq1 protein, [PIN+]. When fused to GFP and overexpressed in [ps-] [PIN+] cells, Sup35 forms fluorescent rings, and cells with these rings bud off [PSI+] daughters. We investigated the effects of over 400 gene deletions on this de novo induction of [PSI+]. Two classes of gene deletions were identified. Class I deletions (bug1Delta, bem1Delta, arf1Delta, and hog1Delta) reduced the efficiency of [PSI+] induction, but formed rings normally. Class II deletions (las17Delta, vps5Delta, and sac6Delta) inhibited both [PSI+] induction and ring formation. Furthermore, class II deletions reduced, while class I deletions enhanced, toxicity associated with the expanded glutamine repeats of the huntingtin protein exon 1 that causes Huntington's disease. This suggests that prion formation and polyglutamine aggregation involve a multi-phase process that can be inhibited at different steps. PMID- 21625619 TI - A -436C>A polymorphism in the human FAS gene promoter associated with severe childhood malaria. AB - Human genetics and immune responses are considered to critically influence the outcome of malaria infections including life-threatening syndromes caused by Plasmodium falciparum. An important role in immune regulation is assigned to the apoptosis-signaling cell surface receptor CD95 (Fas, APO-1), encoded by the gene FAS. Here, a candidate-gene association study including variant discovery at the FAS gene locus was carried out in a case-control group comprising 1,195 pediatric cases of severe falciparum malaria and 769 unaffected controls from a region highly endemic for malaria in Ghana, West Africa. We found the A allele of c. 436C>A (rs9658676) located in the promoter region of FAS to be significantly associated with protection from severe childhood malaria (odds ratio 0.71, 95% confidence interval 0.58-0.88, p(empirical) = 0.02) and confirmed this finding in a replication group of 1,412 additional severe malaria cases and 2,659 community controls from the same geographic area. The combined analysis resulted in an odds ratio of 0.71 (95% confidence interval 0.62-0.80, p = 1.8*10-7, n = 6035). The association applied to c.-436AA homozygotes (odds ratio 0.47, 95% confidence interval 0.36-0.60) and to a lesser extent to c.-436AC heterozygotes (odds ratio 0.73, 95% confidence interval 0.63-0.84), and also to all phenotypic subgroups studied, including severe malaria anemia, cerebral malaria, and other malaria complications. Quantitative FACS analyses assessing CD95 surface expression of peripheral blood mononuclear cells of naive donors showed a significantly higher proportion of CD69+CD95+ cells among persons homozygous for the protective A allele compared to AC heterozygotes and CC homozygotes, indicating a functional role of the associated CD95 variant, possibly in supporting lymphocyte apoptosis. PMID- 21625620 TI - COL4A1 mutations cause ocular dysgenesis, neuronal localization defects, and myopathy in mice and Walker-Warburg syndrome in humans. AB - Muscle-eye-brain disease (MEB) and Walker Warburg Syndrome (WWS) belong to a spectrum of autosomal recessive diseases characterized by ocular dysgenesis, neuronal migration defects, and congenital muscular dystrophy. Until now, the pathophysiology of MEB/WWS has been attributed to alteration in dystroglycan post translational modification. Here, we provide evidence that mutations in a gene coding for a major basement membrane protein, collagen IV alpha 1 (COL4A1), are a novel cause of MEB/WWS. Using a combination of histological, molecular, and biochemical approaches, we show that heterozygous Col4a1 mutant mice have ocular dysgenesis, neuronal localization defects, and myopathy characteristic of MEB/WWS. Importantly, we identified putative heterozygous mutations in COL4A1 in two MEB/WWS patients. Both mutations occur within conserved amino acids of the triple-helix-forming domain of the protein, and at least one mutation interferes with secretion of the mutant proteins, resulting instead in intracellular accumulation. Expression and posttranslational modification of dystroglycan is unaltered in Col4a1 mutant mice indicating that COL4A1 mutations represent a distinct pathogenic mechanism underlying MEB/WWS. These findings implicate a novel gene and a novel mechanism in the etiology of MEB/WWS and expand the clinical spectrum of COL4A1-associated disorders. PMID- 21625621 TI - A deficiency of ceramide biosynthesis causes cerebellar purkinje cell neurodegeneration and lipofuscin accumulation. AB - Sphingolipids, lipids with a common sphingoid base (also termed long chain base) backbone, play essential cellular structural and signaling functions. Alterations of sphingolipid levels have been implicated in many diseases, including neurodegenerative disorders. However, it remains largely unclear whether sphingolipid changes in these diseases are pathological events or homeostatic responses. Furthermore, how changes in sphingolipid homeostasis shape the progression of aging and neurodegeneration remains to be clarified. We identified two mouse strains, flincher (fln) and toppler (to), with spontaneous recessive mutations that cause cerebellar ataxia and Purkinje cell degeneration. Positional cloning demonstrated that these mutations reside in the Lass1 gene. Lass1 encodes (dihydro)ceramide synthase 1 (CerS1), which is highly expressed in neurons. Both fln and to mutations caused complete loss of CerS1 catalytic activity, which resulted in a reduction in sphingolipid biosynthesis in the brain and dramatic changes in steady-state levels of sphingolipids and sphingoid bases. In addition to Purkinje cell death, deficiency of CerS1 function also induced accumulation of lipofuscin with ubiquitylated proteins in many brain regions. Our results demonstrate clearly that ceramide biosynthesis deficiency can cause neurodegeneration and suggest a novel mechanism of lipofuscin formation, a common phenomenon that occurs during normal aging and in some neurodegenerative diseases. PMID- 21625622 TI - Reading with a simulated 60-channel implant. AB - First generation retinal prostheses containing 50-60 electrodes are currently in clinical trials. The purpose of this study was to evaluate the theoretical upper limit (best possible) reading performance attainable with a state-of-the-art 60 channel retinal implant and to find the optimum viewing conditions for the task. Four normal volunteers performed full-page text reading tasks with a low resolution, 60-pixel viewing window that was stabilized in the central visual field. Two parameters were systematically varied: (1) spatial resolution (image magnification) and (2) the orientation of the rectangular viewing window. Performance was measured in terms of reading accuracy (% of correctly read words) and reading rates (words/min). Maximum reading performances were reached at spatial resolutions between 3.6 and 6 pixels/char. Performance declined outside this range for all subjects. In optimum viewing conditions (4.5 pixels/char), subjects achieved almost perfect reading accuracy and mean reading rates of 26 words/min for the vertical viewing window and of 34 words/min for the horizontal viewing window. These results suggest that, theoretically, some reading abilities can be restored with actual state-of-the-art retinal implant prototypes if "image magnification" is within an "optimum range." Future retinal implants providing higher pixel resolutions, thus allowing for a wider visual span might allow faster reading rates. PMID- 21625623 TI - Defective CFTR-dependent CREB activation results in impaired spermatogenesis and azoospermia. AB - Cystic fibrosis (CF) is the most common life-limiting recessive genetic disease among Caucasians caused by mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) with over 95% male patients infertile. However, whether CFTR mutations could affect spermatogenesis and result in azoospermia remains an open question. Here we report compromised spermatogenesis, with significantly reduced testicular weight and sperm count, and decreased cAMP responsive element binding protein (CREB) expression in the testes of CFTR knockout mice. The involvement of CFTR in HCO(3) (-) transport and the expression of the HCO(3) (-) sensor, soluble adenylyl cyclase (sAC), are demonstrated for the first time in the primary culture of rat Sertoli cells. Inhibition of CFTR or depletion of HCO(3) (-) could reduce FSH-stimulated, sAC-dependent cAMP production and phosphorylation of CREB, the key transcription factor in spermatogenesis. Decreased CFTR and CREB expression are also observed in human testes with azoospermia. The present study reveals a previously undefined role of CFTR and sAC in regulating the cAMP-CREB signaling pathway in Sertoli cells, defect of which may result in impaired spermatogenesis and azoospermia. Altered CFTR-sAC-cAMP-CREB functional loop may also underline the pathogenesis of various CF-related diseases. PMID- 21625624 TI - "Studying injured minds" - the Vietnam head injury study and 40 years of brain injury research. AB - The study of those who have sustained traumatic brain injuries (TBI) during military conflicts has greatly facilitated research in the fields of neuropsychology, neurosurgery, psychiatry, neurology, and neuroimaging. The Vietnam Head Injury Study (VHIS) is a prospective, long-term follow-up study of a cohort of 1,221 Vietnam veterans with mostly penetrating brain injuries, which has stretched over more than 40 years. The scope of this study, both in terms of the types of injury and fields of examination, has been extremely broad. It has been instrumental in extending the field of TBI research and in exposing pressing medical and social issues that affect those who suffer such injuries. This review summarizes the history of conflict-related TBI research and the VHIS to date, as well as the vast range of important findings the VHIS has established. PMID- 21625625 TI - Tensor decomposition reveals concurrent evolutionary convergences and divergences and correlations with structural motifs in ribosomal RNA. AB - Evolutionary relationships among organisms are commonly described by using a hierarchy derived from comparisons of ribosomal RNA (rRNA) sequences. We propose that even on the level of a single rRNA molecule, an organism's evolution is composed of multiple pathways due to concurrent forces that act independently upon different rRNA degrees of freedom. Relationships among organisms are then compositions of coexisting pathway-dependent similarities and dissimilarities, which cannot be described by a single hierarchy. We computationally test this hypothesis in comparative analyses of 16S and 23S rRNA sequence alignments by using a tensor decomposition, i.e., a framework for modeling composite data. Each alignment is encoded in a cuboid, i.e., a third-order tensor, where nucleotides, positions and organisms, each represent a degree of freedom. A tensor mode-1 higher-order singular value decomposition (HOSVD) is formulated such that it separates each cuboid into combinations of patterns of nucleotide frequency variation across organisms and positions, i.e., "eigenpositions" and corresponding nucleotide-specific segments of "eigenorganisms," respectively, independent of a-priori knowledge of the taxonomic groups or rRNA structures. We find, in support of our hypothesis that, first, the significant eigenpositions reveal multiple similarities and dissimilarities among the taxonomic groups. Second, the corresponding eigenorganisms identify insertions or deletions of nucleotides exclusively conserved within the corresponding groups, that map out entire substructures and are enriched in adenosines, unpaired in the rRNA secondary structure, that participate in tertiary structure interactions. This demonstrates that structural motifs involved in rRNA folding and function are evolutionary degrees of freedom. Third, two previously unknown coexisting subgenic relationships between Microsporidia and Archaea are revealed in both the 16S and 23S rRNA alignments, a convergence and a divergence, conferred by insertions and deletions of these motifs, which cannot be described by a single hierarchy. This shows that mode-1 HOSVD modeling of rRNA alignments might be used to computationally predict evolutionary mechanisms. PMID- 21625626 TI - Rational design of protein stability: effect of (2S,4R)-4-fluoroproline on the stability and folding pathway of ubiquitin. AB - BACKGROUND: Many strategies have been employed to increase the conformational stability of proteins. The use of 4-substituted proline analogs capable to induce pre-organization in target proteins is an attractive tool to deliver an additional conformational stability without perturbing the overall protein structure. Both, peptides and proteins containing 4-fluorinated proline derivatives can be stabilized by forcing the pyrrolidine ring in its favored puckering conformation. The fluorinated pyrrolidine rings of proline can preferably stabilize either a C(gamma)-exo or a C(gamma)-endo ring pucker in dependence of proline chirality (4R/4S) in a complex protein structure. To examine whether this rational strategy can be generally used for protein stabilization, we have chosen human ubiquitin as a model protein which contains three proline residues displaying C(gamma)-exo puckering. METHODOLOGY/PRINCIPAL FINDINGS: While (2S,4R)-4-fluoroproline ((4R)-FPro) containing ubiquitinin can be expressed in related auxotrophic Escherichia coli strain, all attempts to incorporate (2S,4S)-4-fluoroproline ((4S)-FPro) failed. Our results indicate that (4R)-FPro is favoring the C(gamma)-exo conformation present in the wild type structure and stabilizes the protein structure due to a pre-organization effect. This was confirmed by thermal and guanidinium chloride-induced denaturation profile analyses, where we observed an increase in stability of -4.71 kJ.mol(-1) in the case of (4R)-FPro containing ubiquitin ((4R)-FPro-ub) compared to wild type ubiquitin (wt-ub). Expectedly, activity assays revealed that (4R)-FPro-ub retained the full biological activity compared to wt-ub. CONCLUSIONS/SIGNIFICANCE: The results fully confirm the general applicability of incorporating fluoroproline derivatives for improving protein stability. In general, a rational design strategy that enforces the natural occurring proline puckering conformation can be used to stabilize the desired target protein. PMID- 21625629 TI - Ethical issues in deep brain stimulation. AB - Deep brain stimulation (DBS) is currently used to treat neurological disorders like Parkinson's disease, essential tremor, and dystonia, and is explored as an experimental treatment for psychiatric disorders like major depression and obsessive compulsive disorder. This mini review discusses ethical issues in DBS treatment and research, as they have been discussed in the medical and ethical literature. With regard to DBS treatment, the most important issues are balancing risks and benefits and ensuring respect for the autonomous wish of the patient. This implies special attention to patient selection, psycho-social impact of treatment, effects on personal identity, and treatment of children. Moreover, it implies a careful informed consent process in which unrealistic expectations of patients and their families are addressed and in which special attention is given to competence. In the context of research, the fundamental ethical challenge is to promote high-quality scientific research in the interest of future patients, while at the same time safeguarding the rights and interests of vulnerable research subjects. Several guidelines have been proposed to ensure this. One of the preconditions to further development of responsible and transparent research practices is the establishment of a comprehensive registry. PMID- 21625630 TI - Is attentional blink a byproduct of neocortical attractors? AB - This study proposes a computational model for attentional blink or "blink of the mind," a phenomenon where a human subject misses perception of a later expected visual pattern as two expected visual patterns are presented less than 500 ms apart. A neocortical patch modeled as an attractor network is stimulated with a sequence of 14 patterns 100 ms apart, two of which are expected targets. Patterns that become active attractors are considered recognized. A neocortical patch is represented as a square matrix of hypercolumns, each containing a set of minicolumns with synaptic connections within and across both minicolumns and hypercolumns. Each minicolumn consists of locally connected layer 2/3 pyramidal cells with interacting basket cells and layer 4 pyramidal cells for input stimulation. All neurons are implemented using the Hodgkin-Huxley multi compartmental cell formalism and include calcium dynamics, and they interact via saturating and depressing AMPA/NMDA and GABA(A) synapses. Stored patterns are encoded with global connectivity of minicolumns across hypercolumns and active patterns compete as the result of lateral inhibition in the network. Stored patterns were stimulated over time intervals to create attractor interference measurable with synthetic spike traces. This setup corresponds with item presentations in human visual attentional blink studies. Stored target patterns were depolarized while distractor patterns where hyperpolarized to represent expectation of items in working memory. Simulations replicated the basic attentional blink phenomena and showed a reduced blink when targets were more salient. Studies on the inhibitory effect of benzodiazepines on attentional blink in human subjects were compared with neocortical simulations where the GABA(A) receptor conductance and decay time were increased. Simulations showed increases in the attentional blink duration, agreeing with observations in human studies. In addition, sensitivity analysis was performed on key parameters of the model, including Ca(2+)-gated K(+) channel conductance, synaptic depression, GABA(A) channel conductance and the NMDA/AMPA ratio of charge entry. PMID- 21625631 TI - Immune responses accelerate ageing: proof-of-principle in an insect model. AB - The pathology of many of the world's most important infectious diseases is caused by the immune response. Additionally age-related disease is often attributed to inflammatory responses. Consequently a reduction in infections and hence inflammation early in life has been hypothesized to explain the rise in lifespan in industrialized societies. Here we demonstrate experimentally for the first time that eliciting an immune response early in life accelerates ageing. We use the beetle Tenebrio molitor as an inflammation model. We provide a proof of principle for the effects of early infection on morbidity late in life and demonstrate a long-lasting cost of immunopathology. Along with presenting a proof of-principle study, we discuss a mechanism for the apparently counter-adaptive persistence of immunopathology in natural populations. If immunopathology from early immune response only becomes costly later in life, natural selection on reducing self-harm would be relaxed, which could explain the presence of immune self-harm in nature. PMID- 21625632 TI - Physical confirmation and mapping of overlapping rat mammary carcinoma susceptibility QTLs, Mcs2 and Mcs6. AB - Only a portion of the estimated heritability of breast cancer susceptibility has been explained by individual loci. Comparative genetic approaches that first use an experimental organism to map susceptibility QTLs are unbiased methods to identify human orthologs to target in human population-based genetic association studies. Here, overlapping rat mammary carcinoma susceptibility (Mcs) predicted QTLs, Mcs6 and Mcs2, were physically confirmed and mapped to identify the human orthologous region. To physically confirm Mcs6 and Mcs2, congenic lines were established using the Wistar-Furth (WF) rat strain, which is susceptible to developing mammary carcinomas, as the recipient (genetic background) and either Wistar-Kyoto (WKy, Mcs6) or Copenhagen (COP, Mcs2), which are resistant, as donor strains. By comparing Mcs phenotypes of WF.WKy congenic lines with distinct segments of WKy chromosome 7 we physically confirmed and mapped Mcs6 to ~33 Mb between markers D7Rat171 and gUwm64-3. The predicted Mcs2 QTL was also physically confirmed using segments of COP chromosome 7 introgressed into a susceptible WF background. The Mcs6 and Mcs2 overlapping genomic regions contain multiple annotated genes, but none have a clear or well established link to breast cancer susceptibility. Igf1 and Socs2 are two of multiple potential candidate genes in Mcs6. The human genomic region orthologous to rat Mcs6 is on chromosome 12 from base positions 71,270,266 to 105,502,699. This region has not shown a genome-wide significant association to breast cancer risk in pun studies of breast cancer susceptibility. PMID- 21625633 TI - Dynamical patterns of cattle trade movements. AB - Despite their importance for the spread of zoonotic diseases, our understanding of the dynamical aspects characterizing the movements of farmed animal populations remains limited as these systems are traditionally studied as static objects and through simplified approximations. By leveraging on the network science approach, here we are able for the first time to fully analyze the longitudinal dataset of Italian cattle movements that reports the mobility of individual animals among farms on a daily basis. The complexity and inter relations between topology, function and dynamical nature of the system are characterized at different spatial and time resolutions, in order to uncover patterns and vulnerabilities fundamental for the definition of targeted prevention and control measures for zoonotic diseases. Results show how the stationarity of statistical distributions coexists with a strong and non-trivial evolutionary dynamics at the node and link levels, on all timescales. Traditional static views of the displacement network hide important patterns of structural changes affecting nodes' centrality and farms' spreading potential, thus limiting the efficiency of interventions based on partial longitudinal information. By fully taking into account the longitudinal dimension, we propose a novel definition of dynamical motifs that is able to uncover the presence of a temporal arrow describing the evolution of the system and the causality patterns of its displacements, shedding light on mechanisms that may play a crucial role in the definition of preventive actions. PMID- 21625634 TI - Hepatitis B infection is associated with asymptomatic malaria in the Brazilian Amazon. AB - BACKGROUND: Areas that are endemic for malaria are also highly endemic for hepatitis B virus (HBV) infection. Nevertheless, it is unknown whether HBV infection modifies the clinical presentation of malaria. This study aimed to address this question. METHODOLOGY AND FINDINGS: An observational study of 636 individuals was performed in Rondonia, western Amazon, Brazil between 2006 and 2007. Active and passive case detections identified Plasmodium infection by field microscopy and nested Polymerase Chain Reaction (PCR). HBV infections were identified by serology and confirmed by real-time PCR. Epidemiological information and plasma cytokine profiles were studied. The data were analyzed using adjusted multinomial logistic regression. Plasmodium-infected individuals with active HBV infection were more likely to be asymptomatic (OR: 120.13, P<0.0001), present with lower levels of parasitemia and demonstrate a decreased inflammatory cytokine profile. Nevertheless, co-infected individuals presented higher HBV viremia. Plasmodium parasitemia inversely correlated with plasma HBV DNA levels (r = -0.6; P = 0.0003). CONCLUSION: HBV infection diminishes the intensity of malaria infection in individuals from this endemic area. This effect seems related to cytokine balance and control of inflammatory responses. These findings add important insights to the understanding of the factors affecting the clinical outcomes of malaria in endemic regions. PMID- 21625635 TI - Mesenchymal stem cells in a transgenic mouse model of multiple system atrophy: immunomodulation and neuroprotection. AB - BACKGROUND: Mesenchymal stem cells (MSC) are currently strong candidates for cell based therapies. They are well known for their differentiation potential and immunoregulatory properties and have been proven to be potentially effective in the treatment of a large variety of diseases, including neurodegenerative disorders. Currently there is no treatment that provides consistent long-term benefits for patients with multiple system atrophy (MSA), a fatal late onset alpha-synucleinopathy. Principally neuroprotective or regenerative strategies, including cell-based therapies, represent a powerful approach for treating MSA. In this study we investigated the efficacy of intravenously applied MSCs in terms of behavioural improvement, neuroprotection and modulation of neuroinflammation in the (PLP)-alphasynuclein (alphaSYN) MSA model. METHODOLOGY/PRINCIPAL FINDINGS: MSCs were intravenously applied in aged (PLP)-alphaSYN transgenic mice. Behavioural analyses, defining fine motor coordination and balance capabilities as well as stride length analysis, were performed to measure behavioural outcome. Neuroprotection was assessed by quantifying TH neurons in the substantia nigra pars compacta (SNc). MSC treatment on neuroinflammation was analysed by cytokine measurements (IL-1alpha, IL-2, IL-4, IL-5, IL-6, IL-10, IL-17, GM-CSF, INFgamma, MCP-1, TGF-beta1, TNF-alpha) in brain lysates together with immunohistochemistry for T-cells and microglia. Four weeks post MSC treatment we observed neuroprotection in the SNc, as well as downregulation of cytokines involved in neuroinflammation. However, there was no behavioural improvement after MSC application. CONCLUSIONS/SIGNIFICANCE: To our knowledge this is the first experimental approach of MSC treatment in a transgenic MSA mouse model. Our data suggest that intravenously infused MSCs have a potent effect on immunomodulation and neuroprotection. Our data warrant further studies to elucidate the efficacy of systemically administered MSCs in transgenic MSA models. PMID- 21625636 TI - Design and synthesis of a quintessential self-transmissible IncX1 plasmid, pX1.0. AB - DNA exchange in bacteria via conjugative plasmids is believed to be among the most important contributing factors to the rapid evolution- and diversification rates observed in bacterial species. The IncX1 plasmids are particularly interesting in relation to enteric bacteria, and typically carry genetic loads like antibiotic resistance genes and virulence factors. So far, however, a "pure" version of these molecular parasites, without genetic loads, has yet to be isolated from the environment. Here we report the construction of pX1.0, a fully synthesized IncX1 plasmid capable of horizontal transfer between different enteric bacteria. The designed pX1.0 sequence was derived from the consensus gene content of five IncX1 plasmids and three other, more divergent, members of the same phylogenetic group. The pX1.0 plasmid was shown to replicate stably in E. coli with a plasmid DNA per total DNA ratio corresponding to approximately 3-9 plasmids per chromosome depending on the growth phase of the host. Through conjugation, pX1.0 was able to self-transfer horizontally into an isogenic strain of E. coli as well as into two additional species belonging to the family Enterobacteriaceae. Our results demonstrate the immediate applicability of recent advances made within the field of synthetic biology for designing and constructing DNA systems, previously existing only in silica. PMID- 21625637 TI - Selective serotonin reuptake inhibitors and gastrointestinal bleeding: a case control study. AB - BACKGROUND: Selective serotonin reuptake inhibitors (SSRIs) have been associated with upper gastrointestinal (GI) bleeding. Given their worldwide use, even small risks account for a large number of cases. This study has been conducted with carefully collected information to further investigate the relationship between SSRIs and upper GI bleeding. METHODS: We conducted a case-control study in hospitals in Spain and in Italy. Cases were patients aged >=18 years with a primary diagnosis of acute upper GI bleeding diagnosed by endoscopy; three controls were matched by sex, age, date of admission (within 3 months) and hospital among patients who were admitted for elective surgery for non-painful disorders. Exposures to SSRIs, other antidepressants and other drugs were defined as any use of these drugs in the 7 days before the day on which upper gastrointestinal bleeding started (index day). RESULTS: 581 cases of upper GI bleeding and 1358 controls were considered eligible for the study; no differences in age or sex distribution were observed between cases and controls after matching. Overall, 4.0% of the cases and 3.3% of controls used an SSRI antidepressant in the week before the index day. No significant risk of upper GI bleeding was encountered for SSRI antidepressants (adjusted odds ratio, 1.06, 95% CI, 0.57-1.96) or for whichever other grouping of antidepressants. CONCLUSIONS: The results of this case-control study showed no significant increase in upper GI bleeding with SSRIs and provide good evidence that the magnitude of any increase in risk is not greater than 2. PMID- 21625638 TI - Perceptual training prevents the emergence of the other race effect during infancy. AB - Experience plays a crucial role in the development of the face processing system. At 6 months of age infants can discriminate individual faces from their own and other races. By 9 months of age this ability to process other-race faces is typically lost, due to minimal experience with other-race faces, and vast exposure to own-race faces, for which infants come to manifest expertise [1]. This is known as the Other Race Effect. In the current study, we demonstrate that exposing Caucasian infants to Chinese faces through perceptual training via picture books for a total of one hour between 6 and 9 months allows Caucasian infants to maintain the ability to discriminate Chinese faces at 9 months of age. The development of the processing of face race can be modified by training, highlighting the importance of early experience in shaping the face representation. PMID- 21625639 TI - Sphingosine kinase 1 regulates the Akt/FOXO3a/Bim pathway and contributes to apoptosis resistance in glioma cells. AB - The aim of this study was to investigate the mechanism through which Sphingosine kinase-1 (SPHK1) exerts its anti-apoptosis activity in glioma cancer cells. We here report that dysregulation of SPHK1 alters the sensitivity of glioma to apoptosis both in vitro and in vivo. Further mechanistic study examined the expression of Bcl-2 family members, including Bcl-2, Mcl-1, Bax and Bim, in SPHK1 overexpressing glioma cells and revealed that only pro-apoptotic Bim was downregulated by SPHK1. Moreover, the transcriptional level of Bim was also altered by SPHK1 in glioma cells. We next confirmed the correlation between SPHK1 and Bim expression in primary glioma specimens. Importantly, increasing SPHK1 expression in glioma cells markedly elevated Akt activity and phosphorylated inactivation of FOXO3a, which led to downregulation of Bim. A pharmacological approach showed that these effects of SPHK1 were dependent on phosphatidylinositol 3-kinase (PI3K). Furthermore, effects of SPHK1 on Akt/FOXO3a/Bim pathway could be reversed by SPHK1 specific RNA interference or SPHK1 inhibitor. Collectively, our results indicate that regulation of the Akt/FOXO3a/Bim pathway may be a novel mechanism by which SPHK1 protects glioma cells from apoptosis, thereby involved in glioma tumorigenesis. PMID- 21625640 TI - The central clock neurons regulate lipid storage in Drosophila. AB - A proper balance of lipid breakdown and synthesis is essential for achieving energy homeostasis as alterations in either of these processes can lead to pathological states such as obesity. The regulation of lipid metabolism is quite complex with multiple signals integrated to control overall triglyceride levels in metabolic tissues. Based upon studies demonstrating effects of the circadian clock on metabolism, we sought to determine if the central clock cells in the Drosophila brain contribute to lipid levels in the fat body, the main nutrient storage organ of the fly. Here, we show that altering the function of the Drosophila central clock neurons leads to an increase in fat body triglycerides. We also show that although triglyceride levels are not affected by age, they are increased by expression of the amyloid-beta protein in central clock neurons. The effect on lipid storage seems to be independent of circadian clock output as changes in triglycerides are not always observed in genetic manipulations that result in altered locomotor rhythms. These data demonstrate that the activity of the central clock neurons is necessary for proper lipid storage. PMID- 21625641 TI - Dysfunctional CFTR alters the bactericidal activity of human macrophages against Pseudomonas aeruginosa. AB - Chronic inflammation of the lung, as a consequence of persistent bacterial infections by several opportunistic pathogens represents the main cause of mortality and morbidity in cystic fibrosis (CF) patients. Mechanisms leading to increased susceptibility to bacterial infections in CF are not completely known, although the involvement of cystic fibrosis transmembrane conductance regulator (CFTR) in microbicidal functions of macrophages is emerging. Tissue macrophages differentiate in situ from infiltrating monocytes, additionally, mature macrophages from different tissues, although having a number of common activities, exhibit variation in some molecular and cellular functions. In order to highlight possible intrinsic macrophage defects due to CFTR dysfunction, we have focused our attention on in vitro differentiated macrophages from human peripheral blood monocytes. Here we report on the contribution of CFTR in the bactericidal activity against Pseudomonas aeruginosa of monocyte derived human macrophages. At first, by real time PCR, immunofluorescence and patch clamp recordings we demonstrated that CFTR is expressed and is mainly localized to surface plasma membranes of human monocyte derived macrophages (MDM) where it acts as a cAMP-dependent chloride channel. Next, we evaluated the bactericidal activity of P. aeruginosa infected macrophages from healthy donors and CF patients by antibiotic protection assays. Our results demonstrate that control and CF macrophages do not differ in the phagocytic activity when infected with P. aeruginosa. Rather, although a reduction of intracellular live bacteria was detected in both non-CF and CF cells, the percentage of surviving bacteria was significantly higher in CF cells. These findings further support the role of CFTR in the fundamental functions of innate immune cells including eradication of bacterial infections by macrophages. PMID- 21625642 TI - Mth10b, a unique member of the Sac10b family, does not bind nucleic acid. AB - The Sac10b protein family is regarded as a group of nucleic acid-binding proteins that are highly conserved and widely distributed within archaea. All reported members of this family are basic proteins that exist as homodimers in solution and bind to DNA and/or RNA without apparent sequence specificity in vitro. Here, we reported a unique member of the family, Mth10b from Methanobacterium thermoautotrophicum DeltaH, whose amino acid sequence shares high homology with other Sac10b family proteins. However, unlike those proteins, Mth10b is an acidic protein; its potential isoelectric point is only 4.56, which is inconsistent with the characteristics of a nucleic acid-binding protein. In this study, Mth10b was expressed in Escherichia coli and purified using a three-column chromatography purification procedure. Biochemical characterization indicated that Mth10b should be similar to typical Sac10b family proteins with respect to its secondary and tertiary structure and in its preferred oligomeric forms. However, an electrophoretic mobility shift analysis (EMSA) showed that neither DNA nor RNA bound to Mth10b in vitro, indicating that either Mth10b likely has a physiological function that is distinct from those of other Sac10b family members or nucleic acid-binding ability may not be a fundamental factor to the actual function of the Sac10b family. PMID- 21625643 TI - Self-organized discrimination of resources. AB - When selecting a resource to exploit, an insect colony must take into account at least two constraints: the resource must be abundant enough to sustain the whole group, but not too large to limit exploitation costs, and risks of conflicts with other colonies. Following recent results on cockroaches and ants, we introduce here a behavioral mechanism that satisfies these two constraints. Individuals simply modulate their probability to switch to another resource as a function of the local density of conspecifics locally detected. As a result, the individuals gather at the smallest resource that can host the whole group, hence reducing competition and exploitation costs while fulfilling the overall group's needs. Our analysis reveals that the group becomes better at discriminating between similar resources as it grows in size. Also, the discrimination mechanism is flexible and the group readily switches to a better suited resource as it appears in the environment. The collective decision emerges through the self-organization of individuals, that is, in absence of any centralized control. It also requires a minimal individual cognitive investment, making the proposed mechanism likely to occur in other social species and suitable for the development of distributed decision making tools. PMID- 21625644 TI - Modulation of the CD95-induced apoptosis: the role of CD95 N-glycosylation. AB - Protein modifications of death receptor pathways play a central role in the regulation of apoptosis. It has been demonstrated that O-glycosylation of TRAIL receptor (R) is essential for sensitivity and resistance towards TRAIL-mediated apoptosis. In this study we ask whether and how glycosylation of CD95 (Fas/APO 1), another death receptor, influences DISC formation and procaspase-8 activation at the CD95 DISC and thereby the onset of apoptosis. We concentrated on N glycostructure since O-glycosylation of CD95 was not found. We applied different approaches to analyze the role of CD95 N-glycosylation on the signal transduction: in silico modeling of CD95 DISC, generation of CD95 glycosylation mutants (at N136 and N118), modulation of N-glycosylation by deoxymannojirimycin (DMM) and sialidase from Vibrio cholerae (VCN). We demonstrate that N deglycosylation of CD95 does not block DISC formation and results only in the reduction of the procaspase-8 activation at the DISC. These findings are important for the better understanding of CD95 apoptosis regulation and reveal differences between apoptotic signaling pathways of the TRAIL and CD95 systems. PMID- 21625645 TI - Automated Risk Identification of Myocardial Infarction Using Relative Frequency Band Coefficient (RFBC) Features from ECG. AB - Various structural and functional changes associated with ischemic (myocardial infarcted) heart cause amplitude and spectral changes in signals obtained at different leads of ECG. In order to capture these changes, Relative Frequency Band Coefficient (RFBC) features from 12-lead ECG have been proposed and used for automated identification of myocardial infarction risk. RFBC features reduces the effect of subject variabilty in body composition on the amplitude dependent features. The proposed method is evaluated on ECG data from PTB diagnostic database using support vector machine as classifier. The promising result suggests that the proposed RFBC features may be used in the screening and clinical decision support system for myocardial infarction. PMID- 21625646 TI - Smart sensors and virtual physiology human approach as a basis of personalized therapies in diabetes mellitus. AB - Diabetes mellitus (DM) has a growing incidence and prevalence in modern societies, pushed by the aging and change of life styles. Despite the huge resources dedicated to improve their quality of life, mortality and morbidity rates, these are still very poor. In this work, DM pathology is revised from clinical and metabolic points of view, as well as mathematical models related to DM, with the aim of justifying an evolution of DM therapies towards the correction of the physiological metabolic loops involved. We analyze the reliability of mathematical models, under the perspective of virtual physiological human (VPH) initiatives, for generating and integrating customized knowledge about patients, which is needed for that evolution. Wearable smart sensors play a key role in this frame, as they provide patient's information to the models.A telehealthcare computational architecture based on distributed smart sensors (first processing layer) and personalized physiological mathematical models integrated in Human Physiological Images (HPI) computational components (second processing layer), is presented. This technology was designed for a renal disease telehealthcare in earlier works and promotes crossroads between smart sensors and the VPH initiative. We suggest that it is able to support a truly personalized, preventive, and predictive healthcare model for the delivery of evolved DM therapies. PMID- 21625647 TI - Entropy and Complexity Analyses in Alzheimer's Disease: An MEG Study. AB - Alzheimer's disease (AD) is one of the most frequent disorders among elderly population and it is considered the main cause of dementia in western countries. This irreversible brain disorder is characterized by neural loss and the appearance of neurofibrillary tangles and senile plaques. The aim of the present study was the analysis of the magnetoencephalogram (MEG) background activity from AD patients and elderly control subjects. MEG recordings from 36 AD patients and 26 controls were analyzed by means of six entropy and complexity measures: Shannon spectral entropy (SSE), approximate entropy (ApEn), sample entropy (SampEn), Higuchi's fractal dimension (HFD), Maragos and Sun's fractal dimension (MSFD), and Lempel-Ziv complexity (LZC). SSE is an irregularity estimator in terms of the flatness of the spectrum, whereas ApEn and SampEn are embbeding entropies that quantify the signal regularity. The complexity measures HFD and MSFD were applied to MEG signals to estimate their fractal dimension. Finally, LZC measures the number of different substrings and the rate of their recurrence along the original time series. Our results show that MEG recordings are less complex and more regular in AD patients than in control subjects. Significant differences between both groups were found in several brain regions using all these methods, with the exception of MSFD (p-value < 0.05, Welch's t-test with Bonferroni's correction). Using receiver operating characteristic curves with a leave-one-out cross-validation procedure, the highest accuracy was achieved with SSE: 77.42%. We conclude that entropy and complexity analyses from MEG background activity could be useful to help in AD diagnosis. PMID- 21625648 TI - Digit ratios have poor indicator value in a wild bird population. AB - Early androgen exposure is known to have long-lasting effects on phenotype, behaviour and even fitness, but difficulties in measuring the exposure hinders the study of its importance in evolutionary context. Digit ratios have been highlighted as a potential easy-to-measure indicator of early steroid exposure, as they have been suggested to reflect steroid, mainly testosterone levels during prenatal development. However, evidence for digit ratios reflecting early steroid levels is weak, as experimental studies, especially in wild populations, are scarce. We studied the association between maternally derived yolk androgens and digit ratios (2D:4D, 2D:3D and 3D:4D) using both correlative data and a rather high level of experimental elevation of yolk androgens in a passerine bird, the pied flycatcher (Ficedula hypoleuca). We also examined whether digit ratios have indicator value in an evolutionary context by studying correlations between digit ratios and reproductive traits, secondary sexual traits and exploratory behaviour. We did not find any association between digit ratios and yolk androgen level either in correlative or experimental data. Digit ratios were neither related to any of the reproductive and secondary sexual traits or exploratory behaviour measured. There was, however, a sex difference in 2D:3D and 3D:4D of adult birds (due to second and fourth digits being shorter in females), which was not apparent in fledglings or captivity-raised juveniles. This suggests that either the sex difference may develop as late as during the sexual maturation for breeding. These results indicate that, in this species, digit ratios are not reliable markers of maternally derived yolk androgen exposure and that they bear little relevance as correlates of the adaptive traits we measured. PMID- 21625649 TI - Temperature, age of mating and starvation determine the role of maternal effects on sex allocation in the mealybug Planococcus citri. AB - Environmental effects on sex allocation are common, yet the evolutionary significance of these effects remains poorly understood. Environmental effects might influence parents, such that their condition directly influences sex allocation by altering the relative benefits of producing sons versus daughters. Alternatively, the environment might influence the offspring themselves, such that the conditions they find themselves in influence their contribution to parental fitness. In both cases, parents might be selected to bias their sex ratio according to the prevailing environmental conditions. Here, we consider sex allocation in the citrus mealybug Planococcus citri, a species with an unusual genetic system in which paternal genes are lost from the germline in males. We test environmental factors that may influence either female condition directly (rearing temperature and food restriction) or that may be used as cues of the future environment (age at mating). Using cytological techniques to obtain primary sex ratios, we show that high temperature, older age at mating and starvation all affect sex allocation, resulting in female-biased sex ratios. However, the effect of temperature is rather weak, and food restriction appears to be strongly associated with reduced longevity and a truncation of the usual schedule of male and offspring production across a female's reproductive lifetime. Instead, facultative sex allocation seems most convincingly affected by age at mating, supporting previous work that suggests that social interactions experienced by adult P. citri females are used when allocating sex. Our results highlight that, even within one species, different aspects of the environment may have conflicting effects on sex allocation. PMID- 21625650 TI - Nestmate recognition in social insects: overcoming physiological constraints with collective decision making. AB - Social insects rank among the most abundant and influential terrestrial organisms. The key to their success is their ability to form tightly knit social groups that perform work cooperatively, and effectively exclude non-members from the colony. An extensive body of research, both empirical and theoretical, has explored how optimal acceptance thresholds could evolve in individuals, driven by the twin costs of inappropriately rejecting true nestmates and erroneously accepting individuals from foreign colonies. Here, in contrast, we use agent based modeling to show that strong nestmate recognition by individuals is often unnecessary. Instead, highly effective nestmate recognition can arise as a colony level property from a collective of individually poor recognizers. Essentially, although an intruder can get by one defender when their odor cues are similar, it is nearly impossible to get past many defenders if there is the slightest difference in cues. The results of our models match observed rejection rates in studies of ants, wasps, and bees. We also show that previous research in support of the optimal threshold theory approach to the problem of nestmate recognition can be alternatively viewed as evidence in favor of the collective formation of a selectively permeable barrier that allows in nestmates (at a significant cost) while rejecting non-nestmates. Finally, this work shows that nestmate recognition has a stronger task allocation component than previously thought, as colonies can nearly always achieve perfect nestmate recognition if it is cost effective for them to do so at the colony level. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00265-010-1094-x) contains supplementary material, which is available to authorized users. PMID- 21625651 TI - Within-group behavioral variation promotes biased task performance and the emergence of a defensive caste in a social spider. AB - The social spider Anelosimus studiosus exhibits a behavioral polymorphism where colony members express either a passive, tolerant behavioral tendency (social) or an aggressive, intolerant behavioral tendency (asocial). Here we test whether asocial individuals act as colony defenders by deflecting the suite of foreign (i.e., heterospecific) spider species that commonly exploit multi-female colonies. We (1) determined whether the phenotypic composition of colonies is associated with foreign spider abundance, (2) tested whether heterospecific spider abundance and diversity affect colony survival in the field, and (3) performed staged encounters between groups of A. studiosus and their colony-level predator Agelenopsis emertoni (A. emertoni)to determine whether asocial females exhibit more defensive behavior. We found that larger colonies harbor more foreign spiders, and the number of asocial colony members was negatively associated with foreign spider abundance. Additionally, colony persistence was negatively associated with the abundance and diversity of foreign spiders within colonies. In encounters with a colony-level predator, asocial females were more likely to exhibit escalatory behavior, and this might explain the negative association between the frequency of asocial females and the presence of foreign spider associates. Together, our results indicate that foreign spiders are detrimental to colony survival, and that asocial females play a defensive role in multi-female colonies. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00265-010-1112-z) contains supplementary material, which is available to authorized users. PMID- 21625652 TI - A strategy to obtain axenic cultures of Arthrospira spp. cyanobacteria. AB - A strategy to obtain axenic cultures of the cyanobacterium Arthrospira sp. ('platensis') Lefevre 1963/M-132-1 strain, consisting of a series of physical and chemical procedures, and the application of an optimized pool of antibiotics, is described in this paper. This strategy, which is an inexpensive and fast way to obtain axenic cultures, can be applied to Arthrospira spp. from culture collections or samples from their natural habitats to eliminate a wide spectrum of contaminants. A high alkaline treatment (pH 12, using KOH) of 72 h is a determinant initial procedure applied to eliminate protozoa and Microcystis sp. Bacteria were eliminated by an optimal antibiotic pool treatment, and Chroococcus sp. residuals were discarded by serial dilution. Optimal concentrations of the antibiotics composing the pool were obtained by a 2(4) factorial central composite rotatable design (CCRD) and Response Surface Methodology (RSM), resulting in: ampicillin 61.6 MUg/ml, penicillin 85.8 MUg/ml, cefoxitin 76.9 MUg/ml, and meropenem 38.9 MUg/ml. The results also indicate that cefoxitin was the most effective antibiotic of this pool. After obtaining the axenic culture, identification of Lefevre 1963/M-132-1 strain was performed using amplification and sequencing of the ITS region (including part of 16S rRNA, tRNA Ile, ITS, tRNA Ala and part of 23S rRNA region) and fatty acid composition data. Data base comparison revealed that Lefevre strain is closely related to A. platensis species (99% identity), while fatty acid composition data suggested A. maxima. These seemingly contradictory results are discussed. PMID- 21625654 TI - Morality, science and the law. PMID- 21625653 TI - Fascin 2b is a component of stereocilia that lengthens actin-based protrusions. AB - Stereocilia are actin-filled protrusions that permit mechanotransduction in the internal ear. To identify proteins that organize the cytoskeleton of stereocilia, we scrutinized the hair-cell transcriptome of zebrafish. One promising candidate encodes fascin 2b, a filamentous actin-bundling protein found in retinal photoreceptors. Immunolabeling of zebrafish hair cells and the use of transgenic zebrafish that expressed fascin 2b fused to green fluorescent protein demonstrated that fascin 2b localized to stereocilia specifically. When filamentous actin and recombinant fusion protein containing fascin 2b were combined in vitro to determine their dissociation constant, a K(d)~0.37 uM was observed. Electron microscopy showed that fascin 2b-actin filament complexes formed parallel actin bundles in vitro. We demonstrated that expression of fascin 2b or espin, another actin-bundling protein, in COS-7 cells induced the formation of long filopodia. Coexpression showed synergism between these proteins through the formation of extra-long protrusions. Using phosphomutant fascin 2b proteins, which mimicked either a phosphorylated or a nonphosphorylated state, in COS-7 cells and in transgenic hair cells, we showed that both formation of long filopodia and localization of fascin 2b to stereocilia were dependent on serine 38. Overexpression of wild-type fascin 2b in hair cells was correlated with increased stereociliary length relative to controls. These findings indicate that fascin 2b plays a key role in shaping stereocilia. PMID- 21625655 TI - The young at risk of CVD are the least likely to receive preventive cardiovascular medications in New Zealand. PMID- 21625656 TI - Under-utilisation of preventive medication in patients with cardiovascular disease is greatest in younger age groups (PREDICT-CVD 15). AB - INTRODUCTION: Blood pressure-lowering (BPL) and lipid-lowering (LL) medications together reduce estimated absolute five-year cardiovascular disease (CVD) risk by >40%. International studies indicate that the proportion of people with CVD receiving pharmacotherapy increases with advancing age. AIM: To compare BPL and LL medications, by sociodemographic characteristics, for patients with known CVD in primary care settings. METHODS: The study population included patients aged 35 74 with known CVD assessed in primary care from July 2006 to October 2009 using a web-based computerised decision support system (PREDICT) for risk assessment and management. Clinical data linked anonymously to national sociodemographic and pharmaceutical dispensing databases. Differences in dispensing BPL and LL medications in six months before first PREDICT assessment was analysed according to age, sex, ethnicity and deprivation. RESULTS: Of 7622 people with CVD, 1625 <55 years old, 2862 were women and 4609 lived in deprived areas (NZDep quintiles 4/5). The study population included 4249 European, 1556 Maori, 1151 Pacific and 329 Indian peoples. BPL medications were dispensed to 81%, LL medications to 73%, both BPL and LL medications to 67%, and 87% received either class of medication. Compared with people aged 65-75, people aged 35-44 were 30-40% less likely and those aged 45-54 were 10-15% less likely to be dispensed BPL, LL medications or both. There were minimal differences in likelihood of dispensing according to sex, ethnicity or deprivation. DISCUSSION: BPL and LL medications are under utilised in patients with known CVD in New Zealand. Only two-thirds of patients in this cohort are on both. Younger patients are considerably less likely to be on recommended medications. PMID- 21625657 TI - Language barriers in the community pharmacy: a survey of northern and western Auckland. AB - INTRODUCTION: Community pharmacists play an important role in increasing patient understanding of medication use. Lack of resources to facilitate communication with non-English speaking (NES) patients may be a communication barrier. AIM: To identify obstacles and coping strategies of community pharmacists when counselling NES patients in Auckland's North Shore and West Auckland. METHODS: A cross-sectional survey of 46 community pharmacies in West Auckland and the northern Auckland region was carried out in February 2009. RESULTS: Community pharmacists frequently counsel NES patients (65% reported at least once a week). Use of bilingual staff was the most commonly employed strategy (78% of respondents) to communicate with these customers. Pharmacies that reported serving NES clients at least daily all had bilingual staff, compared with 70% of pharmacies with less frequent NES contact (p=0.017). No pharmacists reported using professional interpreting services. In our sample, telephone interpreting was the most preferred (63% of respondents) method of communicating with such patients, assuming that further services were made available. DISCUSSION: Community pharmacists frequently serve NES patients, with limited access to interpreting services or translated resources. Although pharmacists have, in some way, adapted to the needs of their patients, our survey suggests that accessible professional interpreting services would further improve pharmacist/NES client interaction. PMID- 21625658 TI - Development, validation (diagnostic accuracy) and audit of the Auckland Sleep Questionnaire: a new tool for diagnosing causes of sleep disorders in primary care. AB - INTRODUCTION: Sleep disorders are common in the community and in primary care populations. Epidemiological surveys generally report insomnia rather than specific diagnoses. AIM: Our aim was to develop a questionnaire that could diagnose common sleep disorders in primary care in order to be able to make a diagnosis of primary insomnia by excluding other causes. Having created such a questionnaire, we then validated it (assessed the diagnostic accuracy). METHODS: The questionnaire was developed from the International Classification of Sleep Disorders using the criteria to create operational criteria. This was used in a primary care survey. A sub-sample of 36 primary care patients (aged over 15 years) was chosen to give a spectrum of disorders. A second sample of 85 patients was taken from a sleep disorder private practice to act as an extra test of validity. RESULTS: The response rate was 73% (36/49) for the primary care validation. The sensitivity and specificity of primary insomnia was 0.78 and 0.77, mood disorders 0.67 and 0.97, obstructive sleep apnoea 0.8 and 0.94, delayed sleep phase disorder was 0.8 and 0.97 and for health problems affecting sleep 0.92 and 0.76. There were a wider range of findings in the private practice audit. DISCUSSION: The validity of the Auckland Sleep Questionnaire is promising. The second version of the questionnaire will use this study to improve its functionality. PMID- 21625659 TI - Factors influencing cigarette access behaviour among 14-15-year-olds in New Zealand: a cross-sectional study. AB - INTRODUCTION: Young people access tobacco from both retail and social sources such as family or friends. Both social influences and density of tobacco retail outlets may be associated with frequency of youth smoking. AIM: To update New Zealand data on demographic factors and social influences associated with retail access and social sources. METHODS: The sample consisted of 14-15-year-old New Zealand youth who self-reported as current smokers. Outcome measures were participants' reporting of three different methods of cigarette access. Descriptive data was presented and multiple logistic regressions were used to examine associations between demographic and social influence factors and cigarette sources. RESULTS: Current smoking habits was found to be the strongest predictor of cigarette source, with daily smokers much more likely to report retail purchase than less than monthly smokers (adjusted OR 11.23, 95% CI 10.10 12.47). The second strongest predictor was parental smoking habits-students with both parents smoking being much more likely to obtain from family than students with neither parent (adjusted OR 2.10, 95% CI 1.95-2.26). Socioeconomic status and living in highly populated areas were also factors significantly associated with particular sources of tobacco. DISCUSSION: Though this study is cross sectional, many potential confounders were controlled for, and results are consistent with the notion that financial means and urban proximity to tobacco retailers are enabling some students to use retailers as a cigarette source. Increased taxation and persuading adult family members to quit and to be more possessive about their cigarettes will help protect youth from smoking. PMID- 21625660 TI - The value of te reo in primary care. AB - INTRODUCTION: The influence of indigeneity is widely recognised as a health determinant; however the impact of the utilisation of the indigenous language on health care has not been closely examined. AIM: To explore the Maori language (te reo) as a determinant of health from a Maori patient's perspective. METHODS: Maori patients were recruited through Maori health networks and the snowballing technique. Thirty participants participated in one of three focus group interviews. A semistructured interview explored the utilisation of health services, comfortability with service delivery and perceptions of general practice surgeries' cultural competency. Thematic analysis was utilised to interpret the data. RESULTS: Te reo was recognised as an important cultural competency, noted by participants as contributing to the development of appropriate doctor-patient relationships and their feelings of being valued within a practice. Patient-led use of te reo was identified as most appropriate, an indicator of quality of care. DISCUSSION: The training of primary care staff in te reo should be encouraged. Developed as a competency, this will see primary care settings better able to respond to Maori patients and in turn support Maori health gains. PMID- 21625661 TI - Patients' and health professionals' perceptions of teamwork in primary care. AB - INTRODUCTION: Effective teamwork in primary care settings is integral to the ongoing health of those with chronic conditions. This study compares patient and health professional perceptions about teams, team membership, and team members' roles. This study aimed to test both the feasibility of undertaking a collaborative method of enquiry as a means of investigating patient perceptions about teamwork in the context of their current health care, and also to compare and contrast these views with those of their usual health professionals in New Zealand suburban general practice settings. METHODS: Using a qualitative methodology, 10 in-depth interviews with eight informants at two practices were conducted and data analysed using inductive thematic analysis. FINDINGS: The methodology successfully elicited confidential interviews with both patients and the health professionals providing their care. Perceptions of the perceived value of team care and qualities facilitating good teamwork were largely concordant. Patient and health professionals differed in their knowledge and understanding about team roles and current chronic care programmes, and had differing perceptions about health care team leadership. CONCLUSIONS: This study supports the consensus that team-based care is essential for those with chronic conditions, but suggests important differences between patient and health professional views as to who should be in a health care team and what their respective roles might be in primary care settings. These differences are worthy of further exploration, as a lack of common understanding has the potential to consistently undermine otherwise well-intentioned efforts to achieve best possible health for patients with chronic conditions. PMID- 21625662 TI - Making sense of chronic illness--a therapeutic approach. AB - INTRODUCTION: A diagnosis of any chronic progressive illness can be a traumatic experience. People wonder how they will be able to cope and health care professionals wonder how they can help those so affected. The aim of the study was to discover how people find meaning when they are diagnosed with chronic illness. The research question asked is: How do people make sense of living with chronic progressive illness? METHOD: This is a qualitative study using a phenomenological approach to apply what is learned to developing therapeutic strategies in order to help those so diagnosed to find the meaning they need in order to live with resilience. Semi-structured interviews with seven people were held in order to determine how they cope with living with chronic progressive illness. The results were then used to develop some suggestions for health professionals as they seek to assist people with chronic progressive illness. FINDINGS: All participants displayed much resilience and determination which was found to emerge from three main themes: memory, hope and meaning. Memory was seen to be the link between all the themes. These are described and, arising out of the results of this study, some suggestions are made in order to assist in management. CONCLUSION: It is possible for health care professionals to assist patients to make sense of chronic illness by helping them to view their illness as part of life, and therefore a challenge to be faced rather than seeing life as dominated by illness. PMID- 21625663 TI - Educational needs of practice nurses in mental health. AB - INTRODUCTION: Large numbers of patients see practice nurses (PNs) daily for their health care. Many of these patients will have a mental health need. International research suggests that practice nurses are undertaking mental health assessment and interventions without the requisite skills and knowledge. AIM: To describe the needs of PNs in mental health education and to explore any involvement with patients with mental health concerns. METHODS: Postal survey of PNs in Hawkes Bay and Tairawhiti regions. Analysis was by descriptive, correlation and inferential statistics and content analysis for open questions. RESULTS: Fifty-two respondents completed the survey (response rate 36%) and the results demonstrate that these PNs are caring for patients with an extensive range of mental health concerns daily. Most common are people with depression and anxiety. The nurses perform a variety of mental health interventions such as counselling and advice on medication and have minimal confidence in their skill level. Their expressed learning needs included education on many mental health conditions including suicidal ideation, all types of depression and bipolar disorder, and of therapies such as cognitive behavioural therapy and family therapy. DISCUSSION: PNs require education and support specifically designed to meet their identified needs in mental health to help improve care to patients. This will require collaboration between secondary mental health services, primary mental health nurses and tertiary institutions. With targeted education these nurses should become more confident and competent in their dealings with people who present to their practice with a mental health concern. PMID- 21625664 TI - (Preventing) two birds with one stone: improving vitamin D levels in the elderly. AB - BACKGROUND AND CONTEXT: A majority of adults have sub-optimal vitamin D levels in the winter in southern New Zealand. This is associated with an increased risk of falls and fragility fractures in the elderly, with long-term adverse outcomes likely. Vitamin D supplementation decreases the risks of both falls and fractures. ASSESSMENT OF PROBLEM: An intervention was undertaken by a small urban general practice to increase the number of elderly patients receiving vitamin D supplementation by linking vitamin D prescription to the annual flu vaccination campaign. RESULTS: Uptake of the supplementation was high and costs to the practice low. Thirty-eight patients were identified for whom long-term supplementation with vitamin D was indicated. STRATEGIES FOR IMPROVEMENT: The study could have been strengthened by incorporating a more formal method of evaluating uptake. LESSONS: Encouraging patients to take supplements as a population-based strategy is a realistic intervention, and linking it to the flu vaccination campaign is both seasonally appropriate and efficient. PMID- 21625665 TI - Repeat prescribing--reducing errors. AB - BACKGROUND AND CONTEXT: Prescribing errors account for a significant proportion of overall error in general practice. Repeat prescribing occurs commonly in New Zealand and is a likely cause of error in practice. ASSESSMENT OF PROBLEM: This paper reports on two related aspects of repeat prescribing; an audit of adherence to a repeat prescribing protocol and self-reported repeat prescribing incidents in a network of 97 general practices. RESULTS: The audit of adherence to the repeat prescribing protocol revealed that some issues persist. In particular, prescribing medication outside an approved list and exceeding specified time limits or maximal scripts before clinical review were problematic. Repeat prescribing encompassed a range of departures of process from minor (such as prescription not available on time) to major (wrong medication). Corrective measures highlighted the importance of both the pharmacist and the patient in error detection. STRATEGIES FOR IMPROVEMENT: Repeat prescribing needs to be recognised as a process potentially fraught with error. Effective practice systems, patient involvement and enhanced pharmacy communication are important contributing factors in reducing error. LESSONS: There is need for robust data regarding error rates in prescribing and the impact of changing prescribing protocols on error rates. PMID- 21625666 TI - The triad of retinal haemorrhage, subdural haemorrhage and encephalopathy in an infant unassociated with evidence of physical injury is not the result of shaking, but is most likely to have been caused by a natural disease: Yes. PMID- 21625667 TI - The triad of retinal haemorrhage, subdural haemorrhage and encephalopathy in an infant unassociated with evidence of physical injury is not the result of shaking, but is most likely to have been caused by a natural disease: No. PMID- 21625668 TI - Lemon balm--Melissa officinalis; also known as lemon balm, bee balm, garden balm, Melissa, melissengeist. PMID- 21625669 TI - Pacific peoples: our health and wellbeing. PMID- 21625671 TI - The frail elderly and their bitter pills. PMID- 21625670 TI - Medical ethics: four principles, two decisions, two roles and no reasons. AB - The 'four principle' view of medical ethics has a strong international pedigree. Despite wide acceptance, there is controversy about the meaning and use of the principles in clinical practice as a checklist for moral behaviour. Recent attempts by medical regulatory authorities to use the four principles to judge medical practitioner behaviour have not met with success in clarifying how these principles can be incorporated into a legal framework. This may reflect the philosophical debate about the relationship between law and morals. In this paper, legal decisions from two cases in which general practitioners have been charged with professional shortcomings are discussed. Difficulties with the application of the four principles (autonomy, beneficence, nonmaleficence and justice) to judge medical practitioner behaviour are highlighted. The four principles are relevant to medical practitioner behaviour, but if applied as justifications for disciplinary decisions without explanation, perverse results may ensue. Solutions are suggested to minimise ambiguities in the application of the four principles: adjudicators should acknowledge the difference between professional and common morality and the statutory requirement to give decisions with reasons. PMID- 21625672 TI - Rapid synthesis of flower-like Cu2O architectures in ionic liquids by the assistance of microwave irradiation with high photochemical activity. AB - A novel and facile protocol for the rapid synthesis of flower-like Cu(2)O architectures is reported in the presence of ionic liquid 1-n-butyl-3-methyl imidazolium tetrafluoroborate ([BMIM]BF(4)) with the assistance of microwave irradiation. The hierarchical structures are assembled from many thin nanosheets with tunable sizes by adjusting the amount of [BMIM]BF(4) in the reaction solution. Noticeably, the flower-like Cu(2)O architectures present a high surface area of 65.77 cm(2) g(-1) with a band gap of about 2.25 eV, and exhibit high and stable photochemical activity for the reduction of Cr(VI) to Cr(III) under visible light irradiation. A reasonable model of an absorption and diffusion limited aggregation process is proposed for explaining the possible formation mechanism of the flower-like Cu(2)O. The approach described in this study provides a feasible and rapid method to synthesize flower-like Cu(2)O with a hierarchical structure that is ready for application in the fields of photocatalytic hazard pollutants. PMID- 21625673 TI - Multivariate analysis of emission decay matrices for distinguishing ground state heterogeneity and excited state reactions of tryptophan. AB - The amino acid tryptophan displays emission solvatochromism, an emission maximum that shifts with solvent polarity, which is often used in protein studies to indicate local environment hydrophobicity. Use of tryptophan solvatochromism in time-resolved protein studies has traditionally been complicated due to the undescribed photokinetics that result in a characteristic multiexponential emission decay. For the first time, by application of the photokinetic matrix decomposition (PMD) multivariate curve resolution method to time-resolved emission decay (TRED) data, a distinguishment between ground state heterogeneous (GSH) and excited state reaction (ESR) type photokinetics of tryptophan in solution is made possible. It is found that molecular tryptophan displays two emission spectra that decay independently, suggesting GSH type photokinetics, one at 347 nm with a lifetime of 0.5 ns and one at 363 nm with a lifetime of 3.1 ns. When tryptophan is incorporated into a peptide, mastoparan X, the data similarly contain two emission spectra that decay independently, but are shifted in wavelength. Photobleaching experiments confirm that the PMD method is sensitive to tryptophan emission quenching, and therefore may be applied to determine the photokinetics of tryptophan that occur in proteins. Future applications of PMD analysis of tryptophan TRED data as a bioanalytical tool for further characterizing dynamic protein processes are discussed. PMID- 21625674 TI - Fluorous ligand capture (FLC): a chemoselective solution-phase strategy for isolating 99mTc-labelled compounds in high effective specific activity. AB - A new approach for preparing (99m)Tc-labelled compounds in high effective specific activity was developed by utilizing a novel fluorous ligand capture (FLC) agent and a chemoselective filtration strategy. This paradigm eliminates the need to use HPLC to obtain technetium(I) based molecular imaging probes free from residual precursor. PMID- 21625675 TI - Single-walled carbon nanohorns coated with Fe2O3 as a superior anode material for lithium ion batteries. AB - A novel composite of Fe(2)O(3) and single-walled carbon nanohorns (SWCNHs) was firstly developed via a simple hydrothermal method. As an anode material for lithium ion batteries, a Fe(2)O(3)/SWCNHs composite shows excellent rate performance and cycle stability, even at a high current density of 1000 mA g(-1). PMID- 21625676 TI - Generation of a mesoporous silica MSU shell onto solid core silica nanoparticles using a simple two-step sol-gel process. AB - Silica core-shell nanoparticles with a MSU shell have been synthesized using several non-ionic poly(ethylene oxide) based surfactants via a two step sol-gel method. The materials exhibit a typical worm-hole pore structure and tunable pore diameters between 2.4 nm and 5.8 nm. PMID- 21625677 TI - Fabrication of mesoporous polymer monolith: a template-free approach. AB - Mesoporous polyacrylonitrile (PAN) monolith has been fabricated by a template free approach using the unique affinity of PAN towards a water/dimethyl sulfoxide (DMSO) mixture. A newly developed Thermally Induced Phase Separation Technique (TIPS) has been used to obtain the polymer monoliths and their microstructures have been controlled by optimizing the concentration and cooling temperature. PMID- 21625678 TI - Enhanced stability and activity of Pt-Y alloy catalysts for electrocatalytic oxygen reduction. AB - We report Pt-based alloys with early transition metals. Significant electrocatalysis occurs during oxygen reduction reaction (ORR) at the Pt-Y alloy electrodes, and the extent depends on the alloy composition. The Pt-Y alloy electrode activity is related to the d-band center position, and the lattice strain and stability for oxygen reduction reaction. PMID- 21625679 TI - Calix[4]arene derivative functionalized lanthanide (Eu, Tb) SBA-15 mesoporous hybrids with covalent bonds: assembly, characterization and photoluminescence. AB - Three kinds of novel macrocylic calix[4]arene derivatives functionalized SBA-15 type of mesoporous hybrids (Calix-S15, Calix-NO(2)-S15 and Calix-NH(2)-S15) are synthesized by co-condensation of tetraethoxysilane (TEOS) and modified organic ligand (Calix-Si, Calix-NO(2)-Si and Calix-NH(2)-Si) in the presence of Pluronic P123 surfactant as a template. The structural preservation of these three parent materials is confirmed by FTIR spectra, (29)Si MAS NMR spectra, XRD pattern, and N(2) adsorption-desorption measurements. The ternary mesoporous luminescent hybrids containing Ln(3+) (Eu(3+), Tb(3+)) complexes covalently attached to the functionalized ordered mesoporous SBA-15, which are designated as Ln(Calix S15)phen, Ln(Calix-NO(2)-S15)phen and Ln(Calix-NH(2)-S15)phen, are obtained by introducing lanthanide ions and 1,10-phenanroline into the corresponding parent material via covalent bond assembling methods. XRD pattern, TEM and N(2) adsorption-desorption measurements are employed to characterize the mesostrcture of the resulting lanthanide mesoporous hybrids. The photoluminescent behavior (luminescence, lifetime, quantum efficiency, and energy transfer) for these chemically bonded mesoporous hybrids is studied in detail. Also, their quantum efficiencies are determined, which indicates that the different mesoporous hybrid material systems derived from different functionalized calix[4]arene derivative bridges present different luminescence behavior. PMID- 21625680 TI - Electrochemical stability of self-assembled monolayers on nanoporous Au. AB - Desorption of thiolate self-assembled monolayers (SAMs) seriously limits the fabrication of thiol-based devices. Here we demonstrate that nanoporous Au produced by dealloying Au-Ag alloys exhibits high electrochemical stability against thiolate desorption. Nanoporous Au has many defective sites, lattice strain and residual Ag on the ligament surface. First-principles calculations indicate that these surface aspects increase the binding energy between a SAM and the surface of nanoporous Au. PMID- 21625681 TI - Irreversible solvent-driven conversion in cyanometalate {Fe2Ni}n (n=2, 3) single molecule magnets. AB - Two cyano-bridged single-molecule magnets of {Fe(III)(4)Ni(II)(2)} and {Fe(III)(6)Ni(II)(3)} stoichiometry are described via their magnetic properties described in the frame of geometrical core distortions and orientations of their local anisotropy axes. PMID- 21625682 TI - Phosphites as ligands in ruthenium-benzylidene catalysts for olefin metathesis. AB - The use of phosphites in second generation, ruthenium-based olefin metathesis pre catalysts leads to an improvement in catalyst stability and activity at low catalyst loadings. PMID- 21625683 TI - Synthesis of titania embedded silica hollow nanospheres via sonication mediated etching and re-deposition. AB - Titania embedded silica hollow nanospheres were synthesized from sonication mediated etching and re-deposition of silica/titania core/shell nanospheres. The designed structure of the hollow nanospheres was proved to be a key factor for the charge trapping/detrapping and resulting bistability in non-volatile organic bistable memory devices (OBDs). PMID- 21625684 TI - 3D vision of human lysozyme adsorbed onto a SBA-15 nanostructured matrix. AB - TEM images of human lysozyme loaded on SBA-15 mesoporous silica were obtained through the immunogold staining (IGS) method. IGS is based on the specific interaction between proteins and colloidal gold-conjugated antibodies. Clear evidence that protein molecules are adsorbed both on the external and on the inner pore surface is presented. PMID- 21625685 TI - A luminescent oxygen channeling biosensor that measures small GTPase activation. AB - We established a homogeneous luminescent oxygen channeling sensor for measuring activation states of small GTPases. The assay quantifies activated GTPases in cell lysates, can be applied to different GTPases, and can be used for multiplex screening. The study will provide guidelines for determining activation states of diverse GTPases in various biological contexts. PMID- 21625686 TI - Opportunistic use of tetrachloroaurate photolysis in the generation of reductive species for the production of gold nanostructures. AB - The photolysis of gold salts is rarely viewed as the initiation for gold nanoparticle (AuNP) formation. Yet, photolysis of AuCl(4)(-) generates chlorine atoms whose rich hydrogen transfer chemistry can readily generate strongly reducing radicals. Interesting precursors include hydrogen peroxide, 2-propanol, 1,4-cyclohexadiene and tetrahydrofuran; all of them yield strongly reducing radicals. Further, this group of substrates has been selected because of the innocuous and volatile nature of reagents and products, thus allowing a remarkably clean synthesis of gold nanostructures. In the case of H(2)O(2) the by products are water and oxygen. The methodology reported here opens the door to particles that can be modified in situ or post-synthesis with custom surface covering without concern for chemical debris from the nanostructure synthesis. PMID- 21625688 TI - UV-resistant superhydrophobic BiOCl nanoflake film by a room-temperature hydrolysis process. AB - We report the fabrication of BiOCl nanoflake film by a simple and reliable hydrolysis of BiCl(3) ethanol solution at room temperature. The as-prepared film, when modified with 1H,1H,2H,2H,-perfluorodecyltrichlorosilane (FDTS), exhibits excellent superhydrophobicity and long-term durability under UV light irradiation for the first time. PMID- 21625687 TI - Investigations into whole water, prototropic and amide proton exchange in lanthanide(III) DOTA-tetraamide chelates. AB - Lanthanide(III) chelates of DOTA-tetraamide ligands have been an area of particular interest since the discovery that water exchange kinetics are dramatically affected by the switch from acetate to amide side-chain donors. More recently these chelates have attracted interest as potential PARACEST agents for use in MRI. In this paper we report the results of studies using chemical exchange saturation transfer (CEST) and some more recently reported chelates to re-examine the exchange processes in this class of chelate. We find that the conclusions of Parker and Aime are, for the most part, solid; water exchange is slow and a substantial amount of prototropic exchange occurs in aqueous solution. The extent of prototropic exchange increases as the pH increases above 8, leading to higher relaxivities at high pH. However, amide protons are found to contribute only a small amount to the relaxivity at high pH. PMID- 21625689 TI - Building of a novel Mn12 Single Molecule Magnet by assembly of anisotropic {Mn3(MU3-O)(salox)3}+ triangles. AB - Assembly of triangular {Mn(III)(3)(O)(salox)(3)}(+) fragments mediated by azido ligands, results in the dodecanuclear [Mn(12)O(4)(salox)(12)(N(3))(4)(MeOH)(4)(H(2)O)(2)] complex with S = 8 ground state and SMM response. PMID- 21625690 TI - Peroxidase activity enhancement of horse cytochrome c by dimerization. AB - The peroxidase activity of horse cytochrome c was enhanced by its dimerization, where its Compound III (oxy-form) and Compound I (oxoferryl porphyrin pi-cation radical) species were detected in the reactions with hydrogen peroxide and meta chloroperbenzoic acid, respectively. These results show that oligomeric cytochrome c can contribute as a proapoptotic conformer by the increased peroxidase activity. PMID- 21625691 TI - Electronic structure and absorption spectra of supramolecular complexes of a fullerene crown ether with a pi-extended TTF derivative. AB - The present work is a theoretical investigation on supramolecular complexes of a fullerene crown ether (A and B isomers) with a derivative of pi-extended tetrathiafulvalene (T). The geometry and the electronic structure of seven different conformers of the complex of dibenzo-18-crown-6 ether of fullero-N methylpyrrolidine with a N-benzyl-N-(4-{[9,10-bis(1,3-dithiol-2-ylidene)-9,10 dihydroanthracen-2-yl]ethynyl}benzyl)ammonium cation were determined. We calculated the complexation energies and the absorption spectra, i.e., the lowest 50 excited electronic states of the complexes have been determined at the ground state optimum geometry. All calculations were carried out employing the density functional theory (DFT) and the time-dependent DFT, using the B3LYP, CAM-B3LYP, omegaB97X-D, and M06-2X functionals in conjunction with the 6-31G(d,p) basis set. Various types of van der Waals interactions are observed in the complexes. Conformer complexation energies (CE) range from 2.54 to 2.14 eV in the gas phase and from 1.75 to 1.34 eV in CHCl(3) solvent at the omegaB97X-D/6-31G(d,p)//M06 2X/6-31G(d,p) level of theory. There are three major features at about 390, 330, and 290 nm in the calculated absorption spectra of all the conformers. The major peaks correspond to T->T, T->T/F (electron density in both T and the fullerene F of B) and to T->F transitions, depending on the particular conformer. Other charge transfer T->F transitions are observed close to the T->T transition, indicating the possibility of photoinduced electron transfer in all these complexes. PMID- 21625692 TI - A comparative first-principles study of orbital hybridization in two-dimensional C, Si, and Ge. AB - Information on orbital hybridization is very important to understand the structural, physical, and chemical properties of a material. Results of a comparative first-principles study on the behaviours of orbital hybridization in the two-dimensional single-element phases by carbon, silicon, and germanium are presented. From the well-known three-dimensional hexagonal lonsdaleite structure, in which the atoms are in ideal sp(3)-bonding, the layer spacing along c-axis is gradually stretched to simulate the evolutions of structural and electronic properties from three-dimensional to two-dimensional lattice configurations in the three materials. A turning point of the total system energy due to the sp(3) to sp(2) transition is observed during this process in carbon. In contrast, no such phenomenon is found in silicon and germanium. The differences in electronic structure and bonding behaviour are further examined through comparative investigation of atomic angular-momentum projected density of states and electronic energy band spectrums of these materials. We demonstrate that the valence electronic orbital in the two-dimensional hexagonal crystals of Si and Ge shows sp(3)-like behaviour for the partial hybridization of s and p(z), which leads to their different lattice configurations to graphene. The role of pi-bonds in stabilizing the flat configuration of graphene is also discussed. PMID- 21625693 TI - Time-resolved emission of flavin adenine dinucleotide in water and water-methanol mixtures. AB - Time-resolved fluorescence decay of flavin adenine dinucleotide (FAD) was studied at room temperature in water and water-methanol mixtures by a fluorescence upconversion technique. The observations were focused on the most initial decay phase (200 ps), before the residual fluorescence assumes a single exponential decay, typical for an extended conformation of the fluorophore. Within the first few picoseconds, where most of the electron transfer coupled quenching takes place, the emission decay curves could be fitted by a stretched exponent, compatible with the inhomogeneous distance dependent electron transfer model. This implies that the population of the excited FAD molecules exhibits a large number of non-identical states, each with its own separation between the donor (adenine) and acceptor (isoalloxazine) moieties, having its own rate of electron transfer. To evaluate the distribution of the separation between the donor acceptor pair, we carried out molecular dynamics simulations of closed conformation of the FAD in water and water-methanol mixtures, sampling the structure at 10 fs intervals. The analysis of the dynamics reveals that within the 4 ps time frame, where most of the nonexponential fluorescence relaxation takes place, the relative motion of the donor-acceptor pair is consistent with a one-dimensional Brownian motion, where the diffusion coefficient and the shape of the confining potential well are solvent dependent. The presence of methanol enhances the diffusion constant and widens the width of the potential well. On the basis of these parameters, the relaxation dynamics was accurately reconstructed as an electron transfer reaction in an inhomogeneous system where the reactants are diffusing within the time frame of the observation. PMID- 21625694 TI - Identification of 3,6-di-O-acetyl-1,2,4-O-orthoacetyl-alpha-D-glucopyranose as a direct evidence for the 4-O-acyl group participation in glycosylation. AB - The formation of 3,6-di-O-acetyl-1,2,4-O-orthoacetyl-alpha-D-glucopyranose was observed in the gold(I)-catalyzed glycosidation of peracetyl glucopyranosyl ortho hexynylbenzoate; experiments with substrates bearing deuterium labeled 2-O-acetyl or 4-O-acetyl groups indicated that the orthoacetate was derived from the 4-O acetyl group, which provided a direct evidence for the remote participation of the 4-O-acyl group in glycosylation. PMID- 21625695 TI - Helical periodic mesoporous 1,4-phenylene-silica nanorods with chiral crystalline walls. AB - Helical periodic mesoporous 1,4-phenylene-silica nanorods with chiral crystalline walls were prepared using the self-assembly of a pair of enantiomers as templates. PMID- 21625696 TI - Facile functionalization of polypeptides by thiol-yne photochemistry for biomimetic materials synthesis. AB - Rapid and highly efficient side-chain functionalization of polypeptides was achieved via combination of ring-opening polymerization of a new clickable monomer of gamma-propargyl-L-glutamate N-carboxyanhydride (PLG-NCA) and thiol-yne photochemistry, which provides a convenient and universal route to prepare diverse polypeptide-based biomimetic hybrid materials. PMID- 21625697 TI - Aptamer-based highly sensitive electrochemiluminescent detection of thrombin via nanoparticle layer-by-layer assembled amplification labels. AB - The preparation and use of a new class of signal amplification label, the CdTe quantum dot layer-by-layer assembled polystyrene microbead composite, for amplified ultrasensitive electrochemiluminescent detection of thrombin is described. PMID- 21625698 TI - Polyethylenes bearing a terminal porphyrin group. AB - An alpha-[Cu(II)-porphyrin]-polyethylene was synthesized for the first time using copper catalyzed 1,3-dipolar azide-alkyne Huisgen cycloaddition yielding highly colored moiety-substituted polyethylene. PMID- 21625699 TI - Uncovering the role of the ZnS treatment in the performance of quantum dot sensitized solar cells. AB - Among the third-generation photovoltaic devices, much attention is being paid to the so-called Quantum Dot sensitized Solar Cells (QDSCs). The currently poor performance of QDSCs seems to be efficiently patched by the ZnS treatment, increasing the output parameters of the devices, albeit its function remains rather unclear. Here new insights into the role of the ZnS layer on the QDSC performance are provided, revealing simultaneously the most active recombination pathways. Optical and AFM characterization confirms that the ZnS deposit covers, at least partially, both the TiO(2) nanoparticles and the QDs (CdSe). Photoanodes submitted to the ZnS treatment before and/or after the introduction of colloidal CdSe QDs were studied by electrochemical impedance spectroscopy, cyclic voltammetry and photocurrent experiments. The corresponding results prove that the passivation of the CdSe QDs rather than the blockage of the TiO(2) surface is the main factor leading to the efficiency improvement. In addition, a study of the ultrafast carrier dynamics by means of the Lens-Free Heterodyne Detection Transient Grating technique indicates that the ZnS shell also increases the rate of electron transfer. The dual role of the ZnS layer should be kept in mind in the quest for new modifiers for enhancing the performance of QDSCs. PMID- 21625700 TI - Simulations of photoemission and equilibrium redox processes of ionic liquids: the role of ion pairing and long-range polarization. AB - We have studied oxidation and reduction of ionic liquids using methods of theoretical chemistry. In particular, we have modeled photoelectron spectra of 1 ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ([emim][Tf(2)N]) and 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ([bmim][Tf(2)N]) ionic liquids and their components. We have considered ion pairs in the gas phase and solvated in the water represented by a dielectric continuum. We have also characterized the isolated cation and anion of the ionic liquids. The calculated quantities have been compared with available experiments. The photoelectron spectra were modeled within linearized reflection approximation, using a composite ab initio approach based on combination of the MP2 method for the ground state, the PMP2 method for the first ionized electronic state and the TDDFT method to estimate the higher ionization energies. We have also briefly explored energetics of processes following the vertical electron attachment/detachment. Our calculations were able to reproduce the measured photoelectron spectra. We have shown that the valence photoelectron spectrum results from simultaneous ionization from many molecular orbitals. In the threshold region, the electron is ejected from the anion in the gas phase ion pair while the cationic moiety is ionized when the ion pair is solvated. We have investigated the effect of both short-range and long-range interactions on the ionization characteristics, showing that the long-range polarization of the solvent effectively screens the specific short-range interactions. The large difference between the HOMO-LUMO gap and the electrochemical window has been rationalized in terms of relaxation following the redox processes, i.e. solute and solvent geometric relaxation and reactions, e.g. dimerization of the imidazolium radical formed during the reduction. PMID- 21625701 TI - Phase transitions of an ionic liquid self-assembled monolayer on Au. AB - The properties of a surface modified with an ionic liquid self-assembled monolayer (IL-SAM) can be tuned by simply changing the deposition temperature. Mid-IR, SERS, and molecular modelling demonstrated that 1-(12-mercaptododecyl)-3 methylimidazolium bromide (MDMIBr) exhibited a crystalline monolayer for deposition temperatures below 25 degrees C. Above 25 degrees C, the aliphatic chain collapsed into a disordered conformation. At 40 degrees C, another phase transition occurs due to the imidazolium group tilting parallel to the surface. Consequently, the wettability of IL-SAM was tuned over a broad range of contact angle (from 20 degrees to nearly 40 degrees ) by varying the deposition temperature. Permeation of redox mediators to a Au electrode coated with MDMIBr strongly depends on the net charge of the redox mediator. Electron transfer was excellent for neutral and negatively charged redox mediators on electrodes coated with IL-SAM regardless of deposition temperature. PMID- 21625702 TI - Multipolar symmetric squaraines with large two-photon absorption cross-sections in the NIR region. AB - The two-photon absorption (TPA) properties of two extended symmetric squaraine dyes are thoroughly characterized from the experimental and quantum-chemical point of view. The two molecules are specially engineered to have a multipolar structure with increasing complexity, D-pi-A-pi-D and A'-pi-D-pi-A-pi-D-pi-A', respectively. The experimental TPA spectra, measured by means of the Z-scan technique in the femtoseconds regime, display considerably high values of TPA cross sections (sigma(TPA)) for both molecules. In particular, the squaraine with the more extended structure shows the highest value of sigma(TPA) ever reported for this class of molecules. CIS and TDDFT calculations of the one and two-photon absorption properties are carried out to clarify the origin of the observed TPA properties and fully characterize the electronic properties of these compounds. The calculations, in good agreement with the experimental data, suggest that the origin of this exceptionally high sigma(TPA) can be ascribed to the presence of a peripheral A' group, that increases the density of excited states involved in the TPA process. PMID- 21625703 TI - Profiling the tuneable R-SMS-Phos structure in the rhodium(I)-catalyzed hydrogenation of olefins: the last stand? AB - A diversified family of enantiopure P-stereogenic "R-SMS-Phos" {R-SMS-Phos = 1,2 bis[(o-RO-phenyl)(phenyl)phosphino]ethane} ligands wherein R = branched or heteroatom-substituted alkyl, aralkyl, silyl, acyl, sulfonyl, etc. was screened for the Rh(I)-catalyzed hydrogenation of a representative set of olefinic substrates. This systematic and detailed investigation revealed a marked beneficial impact on enantioselectivity and catalyst activity in comparison to Knowles' ultimate DiPAMP {DiPAMP = 1,2-bis[(o-anisyl)(phenyl)phosphino]ethane} design. Mutant ligands with highly enhanced properties possessing particular features wherein the DiPAMP structure is found embedded were identified. PMID- 21625705 TI - Molecular dynamics simulation study on the isomerization and molecular orientation of liquid crystals formed by azobenzene and (1 cyclohexenyl)phenyldiazene. AB - Atomistic molecular dynamics simulations have are used to investigate the liquid crystal systems based on [4-pentyl-(1-cyclohexenyl)]-(4-cyanophenyl)diazene (5CPDCN) and 4-cyano-4'-pentylazobenzene (5AZCN). The results show the growth process of a nematic phase from a disordered phase. Then the phase transition caused by isomerization reaction is studied based on a temporary modification of the dihedral potential. The properties of 5AZCN and 5CPDCN are compared, showing that the orientation of trans-5CPDCN is more highly ordered than trans-5AZCN. This can be attributed to the more extended dihedral angles phi(2) (i.e. the dihedral angle between the ring system and the terminal chain) in trans-5CPDCN enhance the rod-like conformation of the molecules. The orientational correlation functions g(l)(r) (l = 1, 2) are also calculated, by which we find that both 5CPDCN and 5AZCN systems in nematic phase present parallel and anti-parallel dipole correlations. The anti-parallel dipole correlation is localized for the 5CPDCN system; on the contrary, the parallel dipole correlation is weakly localized for the 5AZCN system. PMID- 21625704 TI - Rapid preparation of triazolyl substituted NH-heterocyclic kinase inhibitors via one-pot Sonogashira coupling-TMS-deprotection-CuAAC sequence. AB - The one-pot, three-component Sonogashira coupling-TMS-deprotection-CuAAC ("click") sequence is the key reaction for the rapid synthesis of triazolyl substituted N-Boc protected NH-heterocycles, such as indole, indazole, 4-, 5-, 6 , and 7-azaindoles, 4,7-diazaindole, 7-deazapurines, pyrrole, pyrazole, and imidazole. Subsequently, the protective group was readily removed to give the corresponding triazolyl derivatives of these tremendously important NH heterocycles. All compounds have been tested in a broad panel of kinase assays. Several compounds, 8f, 8h, 8k, and 8l, have been shown to inhibit the kinase PDK1, a target with high oncology relevance, and thus they are promising lead structures for the development of more active derivatives. The X-ray structure analysis of compound 8f in complex with PDK1 has revealed the detailed binding mode of the molecule in the kinase. PMID- 21625706 TI - Phosphonated calix[4]arene-based amphiphiles as scaffolds for fluorescent nano fibres. AB - p-Phosphonic acid calix[4]arene bearing lower rim O-C(18)H(37) alkyl chains assemble into 6 nm diameter fibres, which deposit from toluene onto mica and graphite, as characterised using Atomic Force Microscopy (AFM). Molecular simulations support a micelle-like arrangement of calixarenes with the alkyl chains directed inwards, and they form a composite material with a fluorescent molecule. PMID- 21625707 TI - A new Kagome lattice coordination polymer based on bismuth and pyridine-2,5 dicarboxylate: structure and photoluminescent properties. AB - A new Kagome lattice topology assembled from ML(4) metal-organic polyhedra prepared using bismuth nitrate and pyridine-2,5-dicarboxylate has been obtained via a solvo-thermal reaction. Bi(pydc)(2).(H(3)O(+))(H(2)O)(0.83) is, to the best of our knowledge, the first example of a bismuth-based coordination polymer to form with a Kagome topology. Its structure and photoluminescence properties are reported. PMID- 21625708 TI - Crystallisation kinetics of some archetypal ionic liquids: isothermal and non isothermal determination of the Avrami exponent. AB - The properties of ionic liquids give rise to applications in diverse technology areas including mechanical engineering, mining, aerospace and defence. The arbitrary physical property that defines an ionic liquid is a melting point below 100 degrees C, and as such, an understanding of crystallisation phenomena is extremely important. This is the first report dealing with the mechanism of crystallisation in ionic liquids. Assuming crystallisation of the ionic liquids is a thermal or mass diffusion-controlled process, the values of the isothermal Avrami exponent obtained from three different ionic liquids with three different anions and cations all indicate that growth occurs with a decreasing nucleation rate (n=1.8-2.2). For one of the ionic liquids it was possible to avoid crystallisation by fast cooling and then observe a devitrification upon heating through the glass transition. The isothermal Avrami exponent of devitrification suggested growth with an increasing nucleating rate (n=4.1), compared to a decreasing nucleation rate when crystallisation occurs on cooling from the melt (n=2.0). Two non-isothermal methods were employed to determine the Avrami exponent of devitrification. Both non-isothermal Avrami exponents were in agreement with the isothermal case (n=4.0-4.15). The applicability of JMAK theory suggests that the nucleation event in the ionic liquids selected is a random stochastic process in the volume of the material. Agreement between the isothermal and non-isothermal techniques for determining the Avrami exponent of devitrification suggests that the pre-exponential factor and the activation energy are independent of thermal history. The heating rate dependence of the glass transition enabled the calculation of the fragility index, which suggests that the ionic liquid is a "strong" glass former. This suggests that the temperature dependence of the rate constant could be close to Arrhenius, as assumed by JMAK theory. More generally, therefore, it can be concluded that there is nothing unusual about the crystallisation mechanism of the ionic liquids studied here. PMID- 21625709 TI - Nature of anion-templated pi(+)-pi(+) interactions. AB - Interaction between positively charged aromatic groups (pi(+)-pi(+)) is characterized by anti-parallel, displaced-stacked structures in the presence of counteranions. Binding energies of pyridinium, N-methylpyridinium and N methylimidazolium dimers are much larger than that of benzene-pyridine (pi-pi) and pyridinium-benzene (pi(+)-pi). Stabilization is attributed to attractive electrostatic interaction with significant dispersion contribution. PMID- 21625710 TI - On the physisorption of water on graphene: a CCSD(T) study. AB - The electronic structure of the zero-gap two-dimensional graphene has a charge neutrality point exactly at the Fermi level that limits the practical application of this material. There are several ways to modify the Fermi-level-region of graphene, e.g. adsorption of graphene on different substrates or different molecules on its surface. In all cases the so-called dispersion or van der Waals interactions can play a crucial role in the mechanism, which describes the modification of electronic structure of graphene. The adsorption of water on graphene is not very accurately reproduced in the standard density functional theory (DFT) calculations and highly-accurate quantum-chemical treatments are required. A possibility to apply wavefunction-based methods to extended systems is the use of local correlation schemes. The adsorption energies obtained in the present work by means of CCSD(T) are much higher in magnitude than the values calculated with standard DFT functional although they agree that physisorption is observed. The obtained results are compared with the values available in the literature for binding of water on the graphene-like substrates. PMID- 21625711 TI - Measuring acetic acid dimer modes by ultrafast time-domain Raman spectroscopy. AB - Acetic acid is capable of forming strong multiple hydrogen bonds and therefore different dimeric H-bonded structures in neat liquid phase and in solutions. The low frequency Raman spectra of acetic acid (neat, in aqueous solution and as a function of temperature) were obtained by ultrafast time and polarization resolved optical Kerr effect (OKE) measurements. Isotropic OKE measurements clearly reveal a specific totally symmetric mode related to the dimeric structure H-bond stretching mode. The effects of isotope substitution, water dilution and temperature on this mode were investigated. These results together with anisotropic OKE measurements and density functional theory calculations for a number of possible dimers provide strong evidence for the cyclic dimer structure being the main structure in liquid phase persisting down to acetic acid concentrations of 10 M. Some information about the dimer structure and concentration dependence was inferred. PMID- 21625712 TI - Aqueous synthesis of CdTe nanocrystals: progresses and perspectives. AB - As an alternative choice to CdSe nanocrystals synthesized in organic phase, CdTe nanocrystals synthesized in aqueous solution, have attracted increasing research interests in recent years. This feature article summarizes the aqueous solution based syntheses, some optoelectronic applications, and bioapplications of CdTe nanocrystals, with emphasis on the recent progresses in related fields. PMID- 21625713 TI - Aqueous polymeric sensors based on temperature-induced polymer phase transitions and solvatochromic dyes. AB - This feature article provides, for the first time, an overview of the research that guided the way from fundamental studies of the thermo-responsive phase separation of aqueous polymer solutions to polymeric sensor systems. The incorporation of solvatochromic dyes into thermoresponsive polymers as well as the concepts of polymeric sensors are presented and discussed in detail. PMID- 21625714 TI - A selective near-infrared fluorescent probe for singlet oxygen in living cells. AB - A selective near-infrared fluorescent probe (His-Cy), which features a fast response to (1)O(2) with high sensitivity and selectivity, was designed, synthesized and applied to bioimaging. PMID- 21625715 TI - Rotational-state-specific guiding of large molecules. AB - A beam of polar molecules can be focused and transported through an ac electric quadrupole guide. At a given ac frequency, the transmission of the guide depends on the mass-to-dipole-moment (m/MU) ratio of the molecular quantum state. Here we present a detailed characterization of the m/MU selector, using a pulsed beam of benzonitrile (C(6)H(5)CN) molecules in combination with rotational quantum state resolved detection. The arrival time distribution as well as the transverse velocity distribution of the molecules exiting the selector are measured as a function of ac frequency. The MU/DeltaMU resolution of the selector can be controlled by the applied ac waveforms and a value of up to 20 can be obtained with the present setup. This is sufficient to exclusively transmit benzonitrile molecules in quantum states with the same m/MU value as the absolute ground state. The operation characteristics of the m/MU selector are in quantitative agreement with the outcome of trajectory simulations. PMID- 21625716 TI - DFT characterization of key intermediates in thiols oxidation catalyzed by amavadin. AB - Amavadin is an unusual octa-coordinated V(IV) complex isolated from Amanita muscaria mushrooms. The outer-sphere catalytic properties of such a complex toward several oxidation reactions are well known. Nevertheless, a remarkable example exists, in which the V(V) (d(0)) oxidized form of amavadin is able to electro-catalyze the oxidation of some thiols to the corresponding disulfides through an inner-sphere mechanism (Guedes da Silva et al. J. Am. Chem. Soc.1996, 118, 7568-7573.) The reaction mechanism implies the formation of an amavadin substrate intermediate, whose half-life is about 0.3 s. By means of Density Functional Theory (DFT) computations and Quantum Theory of Atoms in Molecules (QTAIM) analysis of the electron density, we have first characterized the stereoelectronic features of the V(IV) (inactive) and V(V) (active) states of amavadin. Then, the formation of the V(V) complex with methyl mercaptoacetate (MMA), which has been chosen as a prototypical substrate, has been characterized both thermodynamically and kinetically. DFT results reveal that protonation of V(V) amavadin at a carboxylate oxygen not directly involved in the V coordination, favors MMA binding into the first coordination sphere of vanadium, by substitution of the amavadin carboxylate oxygen with that of the substrate and formation of an S-HO hydrogen bond interaction. The latter interaction can promote SH deprotonation and binding of the thiolate group to vanadium. The kinetic and thermodynamic feasibility of the V(V)-MMA intermediates formation is in agreement, along with electrochemical experimental data, also with the biological role exerted by amavadin. Finally, the presence of an ester functional group as an essential requisite for MMA oxidation has been rationalized. PMID- 21625717 TI - Tandem allylic oxidation-condensation/esterification catalyzed by silica gel: an expeditious approach towards antimalarial diaryldienones and enones from natural methoxylated phenylpropenes. AB - A new one-pot strategy has been developed, wherein abundantly available methoxylated phenylpropenes are directly transformed into corresponding dienones (1,5-diarylpenta-2,4-dien-1-ones) and enones (chalcones and cinnamic esters) via allylic oxidation-condensation or allylic oxidation-esterification sequences. Preliminary antimalarial activity studies of the above synthesized diaryldienones and enones against Plasmodium falciparum (Pf3D7) have shown them to be promising lead candidates for developing newer and economical antimalarial agents. In particular, two enones (12b and 13b) were found to possess comparatively better activity (IC(50) = 4.0 and 3.4 MUM, respectively) than licochalcone (IC(50) = 4.1 MUM), a well known natural antimalarial compound. PMID- 21625718 TI - Receptor design and extraction of inorganic fluoride ion from aqueous medium. AB - A receptor with acidic methylene hydrogens is found to act as an efficient binding mode for F(-). This reagent could as well be used for selective and quantitative extraction of F(-) from the aqueous solution of NaF and sea water. PMID- 21625719 TI - Tuning the moisture stability of metal-organic frameworks by incorporating hydrophobic functional groups at different positions of ligands. AB - The introduction of hydrophobic groups (e.g. methyl) at the most adjacent sites of each and every coordinating nitrogen atom of the bipyridine pillar linker in a carboxylate-based bridging MOF could shield the metal ions from attack by water molecules, and thus enhance the water resistance of the MOF structure significantly. PMID- 21625720 TI - A zinc(II) metal-organic framework based on triazole and dicarboxylate ligands for selective adsorption of hexane isomers. AB - A three-dimensional (3D) metal-organic framework {[Zn(2)(HBDC)(2)(dmtrz)(2)].guest}(n) with pcu net has been solvothermally synthesized, which shows selective adsorption of linear and monobranched hexane isomers over a dibranched one. PMID- 21625721 TI - Designed copper-amine complex as an efficient template for one-pot synthesis of Cu-SSZ-13 zeolite with excellent activity for selective catalytic reduction of NOx by NH3. AB - Low-cost copper-amine complex was rationally designed to be a novel template for one-pot synthesis of Cu-SSZ-13 zeolites. Proper confirmation and appropriate size make this complex fit well with CHA cages as an efficient template. The products exhibit superior catalytic performance on NH(3)-SCR reaction. PMID- 21625722 TI - Investigating N-methoxy-N'-aryl ureas in oxidative C-H olefination reactions: an unexpected oxidation behaviour. AB - Herein, we report a urea derived directing group for mild and highly selective oxidative C-H bond olefination. Subsequent intramolecular Michael addition affords dihydroquinazolinones in good yields. The N-O bond of the urea substrate exhibits superior oxidative behaviour compared to a variety of other external oxidants. PMID- 21625723 TI - Hydrogenation of imino bonds with half-sandwich metal catalysts. AB - Imines can be reduced to afford synthetically important amines via a number of means, of which half-sandwich metal complex-effected reduction has gained particular prominence in the past one decade or so. This Feature Article aims to summarise the progress made with such metal catalysts, placing emphasis on our own work. The article covers transfer hydrogenation and hydrogenation, and finishes with a brief account of catalyst immobilisation and mechanistic understanding. PMID- 21625724 TI - Tropospheric ozone. PMID- 21625725 TI - Prodrug design for the potent cardiovascular agent Nomega-hydroxy-L-arginine (NOHA): synthetic approaches and physicochemical characterization. AB - N(omega)-Hydroxy-L-arginine (NOHA)--the physiological nitric oxide precursor--is the intermediate of NO synthase (NOS) catalysis. Besides the important fact of releasing NO mainly at the NOS-side of action, NOHA also represents a potent inhibitor of arginases, making it an ideal therapeutic tool to treat cardiovascular diseases that are associated with endothelial dysfunction. Here, we describe an approach to impart NOHA drug-like properties, particularly by wrapping up the chemically and metabolically instable N-hydroxyguanidine moiety with different prodrug groups. We present synthetic routes that deliver several more or less highly substituted NOHA derivatives in excellent yields. Versatile prodrug strategies were realized, including novel concepts of bioactivation. Prodrug candidates were primarily investigated regarding their hydrolytic and oxidative stabilities. Within the scope of this work, we essentially present the first prodrug approaches for an interesting pharmacophoric moiety, i.e., N hydroxyguanidine. PMID- 21625726 TI - Nucleobase modification as redox DNA labelling for electrochemical detection. AB - Basic aspects of DNA electrochemistry with a strong focus on the use of modified nucleobases as redox probes for electrochemical bioanalysis are reviewed. Intrinsic electrochemical properties of nucleobases in combination with artificial redox-active nucleobase modifications are frequently applied in this field. Synthetic approaches (both chemical and enzymatic) to base-modified nucleic acids are briefly summarized and their applications in redox labelling are discussed. Finally, analytical applications including DNA hybridization, primer extension, PCR, SNP typing, DNA damage and DNA-protein interaction analysis are presented (critical review, 91 references). PMID- 21625727 TI - Planar chiral (eta5-cyclohexadienyl)- and (eta6-arene)-tricarbonylmanganese complexes: synthetic routes and application. AB - Planar chiral arenetricarbonylchromium complexes have been intensively investigated and they have been applied as valuable building blocks for asymmetric synthesis and as ligands for asymmetric catalysis. In contrast, in the field of the isoelectronic cationic [(eta(6)-arene)Mn(CO)(3)](+) complexes, until these last 10 years, very few studies were published involving nonracemic planar chiral cationic complexes and their potential applications, certainly because of the difficult access to enantiopure starting material. In 2009, however, the discovery of the first resolution of such compounds opened a new area for their application in the field of organic as well as of organometallic enantioselective syntheses. We felt it important to write a review on this subject to give an up to-date summary of the methodologies used to prepare enantiomerically pure planar chiral neutral [(eta(5)-cyclohexadienyl)Mn(CO)(3)] and cationic [(eta(6) arene)Mn(CO)(3)](+) complexes as well as their potential in enantioselective synthesis. PMID- 21625728 TI - Tracking sewage derived contamination in riverine settings by analysis of synthetic surfactants. AB - A study has been made of the presence and reactivity of the most commonly used surfactants, both anionic (linear alkylbenzene sulfonates, LAS, and alkyl ethoxysulfates, AES) and non-ionic (alcohol polyethoxylates, AEOs, and nonylphenol polyethoxylates, NPEOs), in water and surface sediments from the middle stretch of the Guadalete River in SW Spain (12 stations). Average values were between 0.1 and 3.7 mg kg(-1) in sediment, and between 0.2 and 37 MUg L(-1) in water. The sorption of surfactants was dominated by hydrophobic mechanisms, so those homologues having longer alkyl chains (e.g. C(18)AEO) showed higher relative percentages and concentrations in sediments compared with water. Local and sharply higher concentrations of these compounds were observed at three sampling stations (7, 9 and 12), indicating the occurrence of wastewater discharges into the river. By analysing the distributions of different surfactant homologues and their metabolites we were able to distinguish between sewage contamination from sources discharging treated and untreated wastewaters. Upstream (stations 1-2), LAS concentrations were below 30 MUg L(-1) and the composition of their degradation intermediates (sulfophenyl carboxylic acids, SPCs) (160 MUg L(-1)) was dominated by short-chain homologues (C(6)-C(9)SPCs), indicating that the degradation of this surfactant is at an advanced stage. The highest concentration (487 MUg L(-1)) of SPCs was detected near the effluent outlet of a sewage treatment plant (STP) (station 12). Sampling stations (7 and 9) affected by untreated wastewater discharges were the only ones showing the presence of the most reactive and biodegradable SPC isomers and homologues (e.g. C(11)SPC). Here, LAS reached the highest concentration values measured (>2 mg L( 1)), and showed a homologue distribution closer to that of commercial mixtures than LAS found at the other stations. PMID- 21625729 TI - The construction of an individually addressable cell array for selective patterning and electroporation. AB - In basic cell biology research and drug discovery, it is important to rapidly introduce genes, proteins or drug compounds into cells without permanent damage. Here, we report a three dimensional SU-8 micro-well structure sandwiched with an indium tin oxide (ITO) electrode-covered slide from the top and an individually addressable array of microelectrodes on the bottom to allow parallel delivery of exogenous molecules into various cells in a spatially specific manner. A positive dielectrophoretic force was selectively applied by energizing appropriate electrodes to capture the dispersed cells at the bottom electrode, while the micro-wells were designed to confine cells in situ when the positive dielectrophoretic force is removed. The combination of spatial positive dielectrophoresis (pDEP) and micro-wells made it possible to construct cell microarrays with specific patterns. Once the cells become attached to the electrodes, different plasmids can be introduced sequentially for selective electroporation. The present cell arraying-assisted electroporation chip integrates a pDEP-assisted cell positioning function with selective electroporation to provide a simple and efficient method for gene transfer. This platform is ideal for high throughput screening of compounds in parallel and thus holds promise for applications in cellular and molecular research. PMID- 21625731 TI - Pulp tissue from primary teeth: new source of stem cells. AB - SHED (stem cells from human exfoliated deciduous teeth) represent a population of postnatal stem cells capable of extensive proliferation and multipotential differentiation. Primary teeth may be an ideal source of postnatal stem cells to regenerate tooth structures and bone, and possibly to treat neural tissue injury or degenerative diseases. SHED are highly proliferative cells derived from an accessible tissue source, and therefore hold potential for providing enough cells for clinical applications. In this review, we describe the current knowledge about dental pulp stem cells and discuss tissue engineering approaches that use SHED to replace irreversibly inflamed or necrotic pulps with a healthy and functionally competent tissue that is capable of forming new dentin. PMID- 21625732 TI - Effect of incorporation of 2-tert-butylaminoethyl methacrylate on flexural strength of a denture base acrylic resin. AB - Polymethyl methacrylate (PMMA) resins have commonly been used as a denture base material. However, denture bases may act as a reservoir for microorganisms and contribute to oral diseases in denture wearers. It is hypothesized that the 2 tert-butylaminoethyl methacrylate (TBAEMA) incorporated to acrylic resins should have antimicrobial activity related to the presence of amino groups on acrylic resin surface. OBJECTIVES: The objectives of this study were to evaluate the presence of amino groups on acrylic resin surface and the influence on flexural strength after incorporation of TBAEMA. MATERIAL AND METHODS: Six groups were divided according to the concentration of TBAEMA incorporated to acrylic resin (Lucitone 550): 0, 0.5, 1.0, 1.5, 1.75 and 2%. Specimens surface were evaluated by Electron Spectroscopy for Chemical Analysis (ESCA) to detect the presence of amino groups, represented by nitrogen ratios. Flexural strength of the specimens was tested and results were analyzed by ANOVA and Tukey's test (alpha=0.05). RESULTS: Different nitrogen ratios were observed on specimen surfaces: 0, 0.13, 0.74, 0.66, 0.92 and 0.33% for groups 0, 0.5, 1.0, 1.5, 1.75, and 2%, respectively. Significant differences were found for flexural strength (p<0.001). The mean flexural strength values were 98.3+/-3.9, 93.3+/-3.2, 83.9+/-2.1, 82.8+/ 5.2, 71.2+/-5.1 and 17.3+/-3.2 MPa for groups 0, 0.5, 1.0, 1.5, 1.75, and 2%, respectively. CONCLUSION: Within the limitations of this study, the incorporation of TBAEMA results in the presence of the potentially antimicrobial amino groups on specimen surfaces, but affect the flexural strength, depending on the concentration of TBAEMA. PMID- 21625733 TI - Evaluation of the color stability of two techniques for reproducing artificial irides after microwave polymerization. AB - The use of ocular prostheses for ophthalmic patients aims to rebuild facial aesthetics and provide an artificial substitute to the visual organ. Natural intemperate conditions promote discoloration of artificial irides and many studies have attempted to produce irides with greater chromatic paint durability using different paint materials. OBJECTIVES: The present study evaluated the color stability of artificial irides obtained with two techniques (oil painting and digital image) and submitted to microwave polymerization. MATERIAL AND METHODS: Forty samples were fabricated simulating ocular prostheses. each sample was constituted by one disc of acrylic resin N1 and one disc of colorless acrylic resin with the iris interposed between the discs. The irides in brown and blue color were obtained by oil painting or digital image. The color stability was determined by a reflection spectrophotometer and measurements were taken before and after microwave polymerization. Statistical analysis of the techniques for reproducing artificial irides was performed by applying the normal data distribution test followed by 2-way ANOVA and Tukey HSD test (alpha=.05). RESULTS: Chromatic alterations occurred in all specimens and statistically significant differences were observed between the oil-painted samples and those obtained by digital imaging. There was no statistical difference between the brown and blue colors. Independently of technique, all samples suffered color alterations after microwave polymerization. CONCLUSION: The digital imaging technique for reproducing irides presented better color stability after microwave polymerization. PMID- 21625734 TI - Composite resin color stability: influence of light sources and immersion media. AB - OBJECTIVE: This study evaluated the influence of light sources and immersion media on the color stability of a nanofilled composite resin. MATERIAL AND METHODS: Conventional halogen, high-power-density halogen and high-power-density light-emitting diode (LED) units were used. There were 4 immersion media: coffee, tea, Coke(r) and artificial saliva. A total of 180 specimens (10 mm x 2 mm) were prepared, immersed in artificial saliva for 24 h at 37+/-1oC, and had their initial color measured with a spectrophotometer according to the CIELab system. Then, the specimens were immersed in the 4 media during 60 days. Data from the color change and luminosity were collected and subjected to statistical analysis by the Kruskall-Wallis test (p<0.05). For immersion time, the data were subjected to two-way ANOVA test and Fisher's test (p<0.05). RESULTS: High-power-density LED (DeltaE=1.91) promoted similar color stability of the composite resin to that of the tested halogen curing units (Jet Lite 4000 plus--DeltaE=2.05; XL 3000- DeltaE=2.28). Coffee (DeltaE=8.40; DeltaL=-5.21) showed the highest influence on color stability of the studied composite resin. CONCLUSION: There was no significant difference in color stability regardless of the light sources, and coffee was the immersion medium that promoted the highest color changes on the tested composite resin. PMID- 21625735 TI - Dental caries experience, oral health status and treatment needs of dental patients with autism. AB - OBJECTIVES: Autism is a lifelong neurodevelopmental disorder. The aims of this study were to investigate whether children with autism have higher caries prevalence, higher periodontal problems, or more treatment needs than children of a control group of non-autistic patients, and to provide baseline data to enable comparison and future planning of dental services to autistic children. MATERIAL AND METHODS: 61 patients with autism aged 6-16 years (45 males and 16 females) attending Dubai and Sharjah Autism Centers were selected for the study. The control group consisted of 61 non-autistic patients chosen from relatives or friends of autistic patients in an attempt to have matched age, sex and socioeconomic status. Each patient received a complete oral and periodontal examination, assessment of caries prevalence, and caries severity. Other conditions assessed were dental plaque, gingivitis, restorations and treatment needs. Chi-square and Fisher's exact test of significance were used to compare groups. RESULTS: The autism group had a male-to-female ratio of 2.8:1. Compared to controls, children with autism had significantly higher decayed, missing or filled teeth than unaffected patients and significantly needed more restorative dental treatment. The restorative index (RI) and Met Need Index (MNI) for the autistic children were 0.02 and 0.3, respectively. The majority of the autistic children either having poor 59.0% (36/61) or fair 37.8% (23/61) oral hygiene compared with healthy control subjects. Likewise, 97.0% (59/61) of the autistic children had gingivitis. CONCLUSIONS: Children with autism exhibited a higher caries prevalence, poor oral hygiene and extensive unmet needs for dental treatment than non-autistic healthy control group. Thus oral health program that emphasizes prevention should be considered of particular importance for children and young people with autism. PMID- 21625736 TI - Apoptosis and survivability of human dental pulp cells under exposure to Bis-GMA. AB - OBJECTIVE: In the present study, we examined whether 2, 2-bis [4-(2-hydroxy-3 methacryloxypropoxy) phenyl] propane (Bis-GMA) has effects on LSC2 cells, human dental pulp cell line. MATERIAL AND METHODS: The viability, cell cycle, and morphology of LSC2 cells were analyzed after exposure to several different concentrations of Bis-GMA. The recovery of viability of Bis-GMA exposed cells was also analyzed in the condition without Bis-GMA. Further, penetration of Bis-GMA to dentin disc was examined using isocratic high-performance liquid chromatography. RESULTS: There was a concentration-dependent decrease in cell proliferation and an increase in cell number in the sub-G1 population after exposure to Bis-GMA. Furthermore, the cells showed typical characteristics of apoptotic cells after the exposure to high concentration of Bis-GMA. In contrast, cells exposed to lower concentrations of Bis-GMA recovered their viability after being cultured without Bis-GMA. We also found that Bis-GMA is capable of penetrating 1-mm-thick dentin discs, though the penetrated concentration was lower than that showing cytotoxicity. CONCLUSION: These results suggest that Bis GMA has cytotoxic effects, though dental pulp exposed to lower concentrations is able to recover their viability when Bis-GMA is removed. PMID- 21625737 TI - Surface roughness of orthodontic band cements with different compositions. AB - OBJECTIVES: The present study evaluated comparatively the surface roughness of four orthodontic band cements after storage in various solutions. MATERIAL AND METHODS: Eight standardized cylinders were made from 4 materials: zinc phosphate cement (ZP), compomer (C), resin-modified glass ionomer cement (RMGIC) and resin cement (RC). Specimens were stored for 24 h in deionized water and immersed in saline (pH 7.0) or 0.1 M lactic acid solution (pH 4.0) for 15 days. Surface roughness readings were taken with a profilometer (Surfcorder SE1200) before and after the storage period. Data were analyzed by two-way ANOVA and Tukey's test (comparison among cements and storage solutions) or paired t-test (comparison before and after the storage period) at 5% significance level. RESULTS: The values for average surface roughness were statistically different (p<0.001) among cements at both baseline and after storage. The roughness values of cements in a decreasing order were ZP>RMGIC>C>R (p<0.001). After 15 days, immersion in lactic acid solution resulted in the highest surface roughness for all cements (p<0.05), except for the RC group (p>0.05). Compared to the current threshold (0.2 um) related to biofilm accumulation, both RC and C remained below the threshold, even after acidic challenge by immersion in lactic acid solution. CONCLUSIONS: Storage time and immersion in lactic acid solution increased the surface roughness of the majority of the tested cements. RC presented the smoothest surface and it was not influenced by storage conditions. PMID- 21625738 TI - Radiopacity evaluation of Portland and MTA-based cements by digital radiographic system. AB - OBJECTIVE: The aim of the present study was to evaluate the radiopacity of Portland and MTA-based cements using the Digora TM digital radiographic system. MATERIAL AND METHODS: The performed tests followed specification number 57 from the American National Standard Institute/American Dental Association (2000) for endodontic sealing materials. The materials were placed in 5 acrylic plates, especially designed for this experiment, along with a graduated aluminum stepwedge varying from 1 to 10 mm in thickness. The set was radiographed at a 30 cm focus-object distance and with 0.2 s exposure time. After the radiographs were taken, the optical laser readings of radiographs were performed by Digora TM system. Five radiographic density readings were performed for each studied material and for each step of the aluminum scale. RESULTS: White ProRoot MTA (155.99+/-8.04), gray ProRoot MTA (155.96+/-16.30) and MTA BIO (143.13+/-16.94) presented higher radiopacity values (p<0.05), while white non-structural Portland (119.76+/-22.34), gray Portland (109.71+/-4.90) and white structural Portland (99.59+/-12.88) presented lower radiopacity values (p<0.05). CONCLUSIONS: It was concluded that MTA-based cements were the only materials presenting radiopacity within the ANSI/ADA specifications. PMID- 21625739 TI - Effects of silica coating and silane surface conditioning on the bond strength of rebonded metal and ceramic brackets. AB - OBJECTIVE: The aim of this study was to evaluate the effects of tribochemical silica coating and silane surface conditioning on the bond strength of rebonded metal and ceramic brackets. MATERIAL AND METHODS: Twenty debonded metal and 20 debonded ceramic brackets were randomly assigned to receive one of the following surface treatments (n=10 for each group): (1) sandblasting (control); (2) tribochemical silica coating combined with silane. Brackets were rebonded to the enamel surface on the labial and lingual sides of premolars with a light polymerized resin composite. All specimens were stored in distilled water for 1 week and then thermocycled (5,000 cycles) between 5-55oC. Shear bond strength values were measured using a universal testing machine. Student's t-test was used to compare the data (alpha=0.05). Failure mode was assessed using a stereomicroscope, and the treated and non-treated bracket surfaces were observed by scanning electron microscopy. RESULTS: Rebonded ceramic brackets treated with silica coating followed by silanization had significantly greater bond strength values (17.7+/-4.4 MPa) than the sandblasting group (2.4+/-0.8 MPa, P<0.001). No significant difference was observed between the rebonded metal brackets treated with silica coating with silanization (15+/-3.9 MPa) and the sandblasted brackets (13.6+/-3.9 MPa). Treated rebonded ceramic specimens primarily exhibited cohesive failure in resin and adhesive failure at the enamel-adhesive interface. CONCLUSIONS: In comparison to sandblasting, silica coating with aluminum trioxide particles followed by silanization resulted in higher bond strengths of rebonded ceramic brackets. PMID- 21625740 TI - Visibility of the mandibular canal on CBCT cross-sectional images. AB - The identification of the mandibular canal (MC) is an important prerequisite for surgical procedures involving the posterior mandible. Cone beam computed tomography (CBCT) represents an advance in imaging technology, but distinguishing the MC from surrounding structures may remain a delicate task. OBJECTIVES: The aim of this study was to assess the visibility of the MC in different regions on CBCT cross-sectional images. MATERIAL AND METHODS: CBCT cross-sectional images of 58 patients (116 hemi-mandibles) were analyzed, and the visibility of the MC in different regions was assessed. RESULTS: The MC was clearly visible in 53% of the hemi-mandibles. Difficult and very difficult visualizations were registered in 25% and 22% of the hemi-mandibles, respectively. The visibility of the MC on distal regions was superior when compared to regions closer to the mental foramen. No differences were found between edentulous and tooth-bearing areas. CONCLUSIONS: The MC presents an overall satisfactory visibility on CBCT cross sectional images in most cases. However, the discrimination of the canal from its surrounds becomes less obvious towards the mental foramen region when cross sectional images are individually analyzed. PMID- 21625741 TI - Validation of an experimental polyurethane model for biomechanical studies on implant supported prosthesis--tension tests. AB - OBJECTIVES: The complexity and heterogeneity of human bone, as well as ethical issues, frequently hinder the development of clinical trials. The purpose of this in vitro study was to determine the modulus of elasticity of a polyurethane isotropic experimental model via tension tests, comparing the results to those reported in the literature for mandibular bone, in order to validate the use of such a model in lieu of mandibular bone in biomechanical studies. MATERIAL AND METHODS: Forty-five polyurethane test specimens were divided into 3 groups of 15 specimens each, according to the ratio (A/B) of polyurethane reagents (PU-1: 1/0.5, PU-2: 1/1, PU-3: 1/1.5). RESULTS: Tension tests were performed in each experimental group and the modulus of elasticity values found were 192.98 MPa (SD=57.20) for PU-1, 347.90 MPa (SD=109.54) for PU-2 and 304.64 MPa (SD=25.48) for PU-3. CONCLUSION: The concentration of choice for building the experimental model was 1/1. PMID- 21625742 TI - Flexural strength of acrylic resin repairs processed by different methods: water bath, microwave energy and chemical polymerization. AB - Denture fractures are common in daily practice, causing inconvenience to the patient and to the dentists. Denture repairs should have adequate strength, dimensional stability and color match, and should be easily and quickly performed as well as relatively inexpensive. OBJECTIVE: The aim of this study was to evaluate the flexural strength of acrylic resin repairs processed by different methods: warm water-bath, microwave energy, and chemical polymerization. MATERIAL AND METHODS: Sixty rectangular specimens (31x10x2.5 mm) were made with warm water bath acrylic resin (Lucitone 550) and grouped (15 specimens per group) according to the resin type used to make repair procedure: 1) specimens of warm water-bath resin (Lucitone 550) without repair (control group); 2) specimens of warm water bath resin repaired with warm water-bath; 3) specimens of warm water-bath resin repaired with microwave resin (Acron MC); 4) specimens of warm water-bath resin repaired with autopolymerized acrylic resin (Simplex). Flexural strength was measured with the three-point bending in a universal testing machine (MTS 810 Material Test System) with load cell of 100 kgf under constant speed of 5 mm/min. Data were analyzed statistically by Kruskal-Wallis test (p<0.05). RESULTS: The control group showed the best result (156.04+/-1.82 MPa). Significant differences were found among repaired specimens and the results were decreasing as follows: group 3 (43.02+/-2.25 MPa), group 2 (36.21+/-1.20 MPa) and group 4 (6.74+/-0.85 MPa). CONCLUSION: All repaired specimens demonstrated lower flexural strength than the control group. Repairs with autopolymerized acrylic resin showed the lowest flexural strength. PMID- 21625743 TI - Characterization of aqueous silver nitrate solutions for leakage tests. AB - OBJECTIVES: To determine the pH over a period of 168 h and the ionic silver content in various concentrations and post-preparation times of aqueous silver nitrate solutions. Also, the possible effects of these factors on microleakage test in adhesive/resin restorations in primary and permanent teeth were evaluated. MATERIAL AND METHODS: A digital pHmeter was used for measuring the pH of the solutions prepared with three types of water (purified, deionized or distilled) and three brands of silver nitrate salt (Merck, Synth or Cennabras) at 0, 1, 2, 24, 48, 72, 96 and 168 h after preparation, and storage in transparent or dark bottles. Ionic silver was assayed according to the post-preparation times (2, 24, 48, 72, 96, 168 h) and concentrations (1, 5, 25, 50%) of solutions by atomic emission spectrometry. For each sample of each condition, three readings were obtained for calculating the mean value. Class V cavities were prepared with enamel margins on primary and permanent teeth and restored with the adhesive systems OptiBond FL or OptiBond SOLO Plus SE and the composite resin Filtek Z 250. After nail polish coverage, the permanent teeth were immersed in 25% or 50% AgNO3 solution and the primary teeth in 5% or 50% AgNO3 solutions for microleakage evaluation. ANOVA and the Tukey's test were used for data analyses (alpha=5%). RESULTS: The mean pH of the solutions ranged from neutral to alkaline (7.9+/-2.2 to 11.8+/-0.9). Mean ionic silver content differed depending on the concentration of the solution (4.75+/-0.5 to 293+/-15.3 ppm). In the microleakage test, significant difference was only observed for the adhesive system factor (p=0.000). CONCLUSIONS: Under the tested experimental conditions and based on the obtained results, it may be concluded that the aqueous AgNO3 solutions: have neutral/alkaline pH and service life of up to 168 h; the level of ionic silver is proportional to the concentration of the solution; even at 5% concentration, the solutions were capable of indicating loss of marginal seal in the composite restorations; the 3-step conventional adhesive system had better performance regarding microleakage in enamel on primary and permanent teeth. PMID- 21625744 TI - Experimental alveolitis in rats: microbiological, acute phase response and histometric characterization of delayed alveolar healing. AB - The pathogenesis of alveolitis is not well known and therefore experimental situations that mimic some features of this disease should be developed. OBJECTIVE: In this study, the evolution of the experimentally induced infection in rat sockets is characterized, which leads to clinical signs of suppurative alveolitis with remarkable wound healing disturbs. MATERIAL AND METHODS: Non infected (Group I) and experimentally infected sockets in Rattus novergicus (Group II) were histometrically evaluated regarding the kinetics of alveolar healing. In addition, the characterization of the present bacteria in inoculation material and the serum levels of C-reactive protein (CRP) were performed. The detected species were Capnocytophaga ochracea, Fusobacterium nucleatum ss nucleatum, Prevotella melaninogenica, Streptococcus anginosus, Treponema socranskii and Streptococcus sanguis. RESULTS: All experimentally infected rats developed suppurative alveolitis, showing higher levels of CRP in comparison to those non-infected ones. Furthermore, infected rats presented a significant delayed wound healing as measured by the histometric analysis (higher persistent polymorphonuclear infiltrate and lower density of newly formed bone). CONCLUSION: These findings indicate that rat sockets with experimentally induced infection produced higher levels of serum CRP, showing the potential of disseminated infection and a disturb in the alveolar repair process in an interesting experimental model for alveolitis studies. PMID- 21625745 TI - Cell block technique as a complementary method in the clinical diagnosis of cyst like lesions of the jaw. AB - OBJECTIVE: To evaluate the applicability of the cell block technique as a complementary method for presumptive diagnosis in the analysis of cyst-like aspirates from jaw lesions prior to histopathological diagnosis. MATERIAL AND METHODS: The sample was made up of 17 cyst-like jaw lesions. After puncture, the aspirates were centrifuged, fixed in formalin, embedded in paraffin and processed. All lesions were biopsied and submitted to histopathological examination. RESULTS: In 9 cases, the cytopathological analysis using the cell block method showed a predominant presence of erythrocytes, inflammatory cells and few epithelial cells. In the other 8 cases, the cell block technique demonstrated the presence of parakeratin, whose histopathological analysis confirmed the occurrence of keratocystic odontogenic tumors (KOTs). CONCLUSIONS: According to the studied cases, the cell block method was proven to be a simple, fast and low-cost technique to effectively differentiate KOTs from other lesions with similar clinical and radiographic features. The cell block technique comprises cellular preparations useful to determine a clinical diagnosis and help to develop a therapeutic plan for those lesions. PMID- 21625746 TI - Effect of gender, facial dimensions, body mass index and type of functional occlusion on bite force. AB - OBJECTIVE: Some factors such as gender, age, craniofacial morphology, body structure, occlusal contact patterns may affect the maximum bite force. Thus, the purposes of this study were to determine the mean maximum bite force in individuals with normal occlusion, and to examine the effect of gender, facial dimensions, body mass index (BMI), type of functional occlusion (canine guidance and group function occlusion) and balancing side interferences on it. MATERIAL AND METHODS: Thirty-four individuals aged 19-20 years-old were selected for this study. Maximum bite force was measured with strain-gauge transducers at first molar region. Facial dimensions were defined by standardized frontal photographs as follows: anterior total facial height (ATFH), bizygomathic facial width (BFW) and intergonial width (IGW). BMI was calculated using the equation weight/height2. The type of functional occlusion and the balancing side interferences of the subjects were identified by clinical examination. RESULTS: Bite force was found to be significantly higher in men than women (p<0.05). While there was a negative correlation between the bite force and ATFH/BFW, ATFH/IGW ratios in men (p<0.05), women did not show any statistically significant correlation (p>0.05). BMI and bite force correlation was not statistically significant (p>0.05). The average bite force did not differ in subjects with canine guidance or group function occlusion and in the presence of balancing side interferences (p>0.05). CONCLUSIONS: Data suggest that bite force is affected by gender. However, BMI, type of functional occlusion and the presence of balancing side interferences did not exert a meaningful influence on bite force. In addition, transverse facial dimensions showed correlation with bite force in only men. PMID- 21625747 TI - Histologic grading and nucleolar organizer regions in oral squamous cell carcinomas. AB - OBJECTIVE: The purposes of this study were to histologically assess different types of oral squamous cell carcinoma and the silver-binding nucleolar organizer region (AgNOR) morphology in neoplastic cells, as well as to quantify the number of AgNORs in each type of carcinoma in order to relate AgNOR count and histologic grading. MATERIAL AND METHODS: Twenty-eight cases of oral squamous cell carcinoma were divided into 4 groups, namely well-differentiated, moderately differentiated, poorly differentiated, and undifferentiated. For NOR study, 3-um thick sections were stained with 50% aqueous silver nitrate solution. The predominant microscopic pattern of NORs was determined. Quantitative analyses of NORs were obtained of all cells present on each histological field using a 0.025 mm2 eyepiece graticule. Different histological fields were analyzed until the total number of NORs was 120 cells for each tumor. Kruskall-Wallis test was applied to compare the groups of sample data at a significance level of p=0.05. RESULTS: The mean number of AgNORs per nucleus was 3.20 for the well differentiated group, 5.33 for the moderately differentiated one, 8.27 for the poorly differentiated one, and 10.08 for the undifferentiated one. AgNOR count was significantly different (p<0.05) among all of the studied groups. CONCLUSION: AgNOR staining technique seems to be a useful diagnostic tool since differences in AgNOR numeric values can be identified in the different types of oral squamous cell carcinoma. This technique is easy to handle and inexpensive, thus justifying its large use in histopathology. PMID- 21625748 TI - Effect of reduced exposure times on the cytotoxicity of resin luting cements cured by high-power led. AB - OBJECTIVE: Applications of resin luting agents and high-power light-emitting diodes (LED) light-curing units (LCUs) have increased considerably over the last few years. However, it is not clear whether the effect of reduced exposure time on cytotoxicity of such products have adequate biocompatibility to meet clinical success. This study aimed at assessing the effect of reduced curing time of five resin luting cements (RLCs) polymerized by high-power LED curing unit on the viability of a cell of L-929 fibroblast cells. MATERIAL AND METHODS: Disc-shaped samples were prepared in polytetrafluoroethylene moulds with cylindrical cavities. The samples were irradiated from the top through the ceramic discs and acetate strips using LED LCU for 20 s (50% of the manufacturer's recommended exposure time) and 40 s (100% exposure time). After curing, the samples were transferred into a culture medium for 24 h. The eluates were obtained and pipetted onto L-929 fibroblast cultures (3x10(4) per well) and incubated for evaluating after 24 h. Measurements were performed by dimethylthiazol diphenyltetrazolium assay. Statistical significance was determined by two-way ANOVA and two independent samples were compared by t-test. RESULTS: Results showed that eluates of most of the materials polymerized for 20 s (except Rely X Unicem and Illusion) reduced to a higher extent cell viability compared to samples of the same materials polymerized for 40 s. Illusion exhibited the least cytotoxicity for 20 s exposure time compared to the control (culture without samples) followed by Rely X Unicem and Rely X ARC (90.81%, 88.90%, and 83.11%, respectively). For Rely X ARC, Duolink and Lute-It 40 s exposure time was better (t=-1.262 p=0,276; t=-9.399 p=0.001; and t=-20.418 p<0.001, respectively). CONCLUSION: The results of this study suggest that reduction of curing time significantly enhances the cytotoxicity of the studied resin cement materials, therefore compromising their clinical performance. PMID- 21625749 TI - Multicenter studies to shed light on fibromuscular displasia and cervical artery dissection. PMID- 21625750 TI - Global aphasia as a predictor of mortality in the acute phase of a first stroke. AB - OBJECTIVE: To establish whether vascular aphasic syndromes can predict stroke outcomes. METHOD: Thirty-seven adults were evaluated for speech and language within 72 hours after a single first-ever ischemic brain lesion, in blind association to CT and/or MR. RESULTS: Speech or language disabilities were found in seven (87.5%) of the eight deceased patients and twenty-six (89.7%) of the twenty-nine survivors. Global aphasia was identified in eleven patients, all with left hemisphere lesions (nine mute; five deceased), consisting on a risk factor for death in the acute stroke phase (rho=0.022). Age (z=1.65; rho>0.09), thrombolysis (rho=0.591), infarct size (rho=0.076) and side (rho=0.649) did not significantly influence survival. Absence of aphasia did not predict a better evolution, regardless of the affected hemisphere. Prevalence of cardiovascular risk factors was similar for all patient groups. CONCLUSION: Global aphasia in acute stroke can adversely affect prognosis, translated into impairment of dominant perisylvian vascular territories, with mutism as an important semiological element. PMID- 21625751 TI - Association study between genetic monoaminergic polymorphisms and OCD response to clomipramine treatment. AB - In the present paper, we investigated the 5HTTLPR and STin2 polymorphisms in the promoter region of the serotonin transporter gene (SLC6A4), the G861C polymorphism (rs6296) of the serotonin receptor 1D beta (HTR1B), the T102C (rs6113) and C516T (rs6305) polymorphisms of the serotonin receptor gene subtype 2A (HTR2A), the DAT UTR, DAT intron 8 and DAT intron 14 of the dopamine transporter gene (SLC6A3), the Val-158-Met (rs4680) polymorphism of the COMT and the silent mutation G1287A (rs5569) in the norepinephrine transporter gene (SLC6A2). We genotyped 41 obsessive-compulsive disorder (OCD) outpatients, classified as good-responders (n=27) and poor-responders (n=14) to treatment with clomipramine according to the Yale Brown Obsessive-Compulsive Scale (YBOCS). Patients who achieved a reduction in symptoms of 40% or more in YBOCS after 14 weeks of treatment were considered good-responders. Genotypes and alleles distribution of the investigated polymorphisms were compared between both groups. We did not find association between the studied polymorphisms and clomipramine response in our sample. PMID- 21625752 TI - Autosomal recessive spastic ataxia of Charlevoix-Saguenay (ARSACS): typical clinical and neuroimaging features in a Brazilian family. AB - Autosomal recessive spastic ataxia of Charlevoix-Saguenay (ARSACS) is a neurodegenerative disorder characterized by late-infantile onset spastic ataxia and other neurological features. ARSACS has a high prevalence in northeastern Quebec, Canada. Several ARSACS cases have been reported outside Canada in recent decades. This is the first report of typical clinical and neuroimaging features in a Brazilian family with probable diagnosis of ARSACS. PMID- 21625753 TI - Brain stem and cerebellum volumetric analysis of Machado Joseph disease patients. AB - Machado-Joseph disease, or spinocerebellar ataxia type 3(MJD/SCA3), is the most frequent late onset spinocerebellar ataxia and results from a CAG repeat expansion in the ataxin-3 gene. Previous studies have found correlation between atrophy of cerebellum and brainstem with age and CAG repeats, although no such correlation has been found with disease duration and clinical manifestations. In this study we test the hypothesis that atrophy of cerebellum and brainstem in MJD/SCA3 is related to clinical severity, disease duration and CAG repeat length as well as to other variables such as age and ICARS (International Cooperative Ataxia Rating Scale). Whole brain high resolution MRI and volumetric measurement with cranial volume normalization were obtained from 15 MJD/SCA3 patients and 15 normal, age and sex-matchedcontrols. We applied ICARS and compared the score with volumes and CAG number, disease duration and age. We found significant correlation of both brain stem and cerebellar atrophy with CAG repeat length, age, disease duration and degree of disability. The Spearman rank correlation was stronger with volumetric reduction of the cerebellum than with brain stem. Our data allow us to conclude that volumetric analysis might reveal progressive degeneration after disease onset, which in turn is linked to both age and number of CAG repeat expansions in SCA 3. PMID- 21625754 TI - Quantitative electroencephalography power and coherence measurements in the diagnosis of mild and moderate Alzheimer's disease. AB - OBJECTIVE: To evaluate the contribution of quantitative electroencephalographic (qEEG) analyses in the diagnosis of Alzheimer's disease (AD). METHOD: Thirty-five patients from the Neurology Outpatients Clinic of PUC-Campinas, diagnosed with AD according to the NINCDS/ADRDA were evaluated, and compared with a control group consisting of 30 individuals with no cognitive deficit. The procedures consisted of clinical-neurological, cognitive and behavioral analyses and the qEEG (absolute power and coherence). RESULTS: The AD group presented greater absolute power values in the delta and theta bands, greater theta/alpha indices and less frontal alpha and beta coherence. Logistic multiple regression models were constructed and those only showing variations in the qEEG (frontal alpha coherence and left frontal absolute theta power) showed an accuracy classification (72.3%) below that obtained in the mini-mental state examination (93%). CONCLUSION: The study of coherence and power in the qEEG showed a relatively limited accuracy with respect to its application in routine clinical practice. PMID- 21625755 TI - gamma band oscillations in parietooccipital areas during performance of a sensorimotor integration task: a qEEG coherence study. AB - This study aimed to elucidate cortical mechanisms involved in anticipatory actions when 23 healthy right-handed subjects had to catch a free falling object through quantitative electroencephalogram (qEEG). For this reason, we used coherence that represents a measurement of linear covariation between two signals in the frequency domain. In addition, we investigated gamma-band (30-100 Hz) activity that is related to cognitive and somatosensory processes. We hypothesized that gamma coherence will be increase in both parietal and occipital areas during moment after ball drop, due to their involvement in manipulation of objects, visuospatial processing, visual perception, stimuli identification and attention processes. We confirmed our hypothesis, an increase in gamma coherence on P3-P4 (t= -2.15; p=0.033) and PZ-OZ (t= -2.16; p=0.034) electrode pairs was verified for a paired t-test. We conclude that to execute tasks involving anticipatory movements (feedforward mechanisms), like our own task, probably, there is no need of a strong participation of visual areas in the process of information organization to manipulate objects and to process visuospatial information regarding the contact hand-object. PMID- 21625756 TI - Cross-cultural translation of the INSPIRIT-R for Brazil and its applicability among epilepsy patients. AB - OBJECTIVE: To describe the cross-cultural adaptation of the INSPIRIT-R instrument for evaluation of religious and spiritual experiences into a Brazilian Portuguese version and its applicability among epileptic patients. METHOD: After the translation and back-translation phases, a multidisciplinary committee compared the back-translation with the original text in order to evaluate its content, comprehensibility, conceptual equivalence, cultural and contextual adjustment for Brazilian population. Lastly, the final version was tested on 50 long-term followed-up outpatients with a confirmed diagnosis of epilepsy in Florianopolis, SC, Brazil. RESULTS: The patients' mean age was 33.7 years (18-55) and 26 (52%) were women. They had attended school for a mean of 8.0 years (3-17) years. Most of them (80%) were Catholics and 82% had a confirmed diagnosis of temporal lobe epilepsy. In the final Portuguese version, questions 3, 7C and 7E required slight modifications, along with the layout of question 7. CONCLUSION: The Brazilian Portuguese version of the INSPIRIT-R instrument was easily understood by most of the patients, after minimal modifications. PMID- 21625757 TI - Cross-cultural adaptation of the Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS) to Brazilian Portuguese. AB - Cervical dystonia (CD) is a prevalent and incapacitating movement disorder which needs a thorough clinical evaluation of every patient to better tailor treatment strategies. In Brazil, there are no validated CD scales that measure the burden of dystonia. The aim of our study was to translate and adapt the Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS) to Brazilian Portuguese. After translation and back-translation according to international methods, a pre-test was carried out with 30 patients. Patients under 8 years of formal schooling had severe difficulty in understanding the whole scale. The scale went through a remodeling process, without loss of its conceptual and semantic properties. The new scale was tested in 15 patients, with good understanding scores. We are now in the process of validation of the adapted scale. PMID- 21625758 TI - Validation of the EuroQol quality of life questionnaire on stroke victims. AB - OBJECTIVE: To validate a quality of life scale, EuroQoL, on stroke patients. METHOD: 67 patients were scored simultaneously for EuroQoL-5 Dimensions (EQ-5D), NIH Stroke Scale (NIHSS) and modified Barthel Index (mBI). Pearson test was used to correlate each scale. Additionally, 31 patients were examined by two independent evaluators on the same day through application of EQ-5D. Kappa statistics were used to evaluate interobserver agreement. RESULTS: EQ-5D showed good correlation with both stroke severity (NIHSS, r= -0.404, P<0.001) and degree of impairment on activities of daily living (mBI, r=0.512, P<0.001). We noticed a good interobserver agreement (k>0.60) in all dimensions evaluated (P<0.01). CONCLUSION: We demonstrated that EQ-5D is reproducible and valid on evaluation of quality of life in patients post stroke in Brazil. PMID- 21625759 TI - Odors as triggering and worsening factors for migraine in men. AB - OBJECTIVE: To assess the role of odors in triggering or worsening migraine in men. METHOD: Ninety-eight male migraineurs from the general population were assessed individually through questionnaires. Environmental factors relating to their migraine were reported, with special focus on the role of odors. RESULTS: Odors were the second most frequent triggering factor for migraine attacks (48%), behind stressful situations (59%). Likewise, odors were the second most frequent worsening factor (73%), just behind excessive light (74%). Thirty-three individuals (33.4%) stated that odors were both triggering and worsening factors for their migraine attacks. Perfume, cigarette smoke and cleaning products were the most frequent migraine-related odors reported by these male migraineurs. CONCLUSION: This was the first study to assess the role of odors in migraine exclusively in men. There was a high degree of odor-related migraine among these men, thus suggesting that patient education could alert such individuals to gender-related factors, since different triggering and worsening factors have been reported by males and females. PMID- 21625760 TI - Extraneural metastases in medulloblastoma. AB - Medulloblastoma is the most common childhood malignant tumor of central nervous system, but it may also occur in adults. It presents high invasive growth with spreading of tumor cells into the leptomeningeal space along the neuroaxis early in the course of the disease. Extraneural metastases are rare but frequently lethal, occurring only in 1 to 5% of patients, and are related, in the most of cases, to the presence of ventriculoperitoneal shunt. Here we characterize the clinical profile of five cases of medulloblastoma with systemic spreading of tumor cells, also comparing them to cases already described in the literature. PMID- 21625761 TI - Epilepsy and occupational accidents in Brazil: a national statistics study. AB - Epilepsy may restrict the patient's daily life. It causes lower quality of life and increased risk for work-related accidents (WRA). The aim of this study is to analyze the implantation of the Epidemiologic and Technical Security System Nexus (ETSSN) and WRA patterns among patients with epilepsy. Data regarding WRA, between 1999 and 2008, on the historical database of WRA Infolog Statistical Yearbook from Brazilian Ministry of Social Security were reviewed. There was a significant increase of reported cases during the ten year period, mainly after the establishment of the ETSSN. The increased granted benefits evidenced the epidemiologic association between epilepsy and WRA. ETSSN possibly raised the registration of occupational accidents and granted benefits. However, the real number of WRA may remain underestimated due to informal economy and house workers' accidents which are usually not included in the official statistics in Brazil. PMID- 21625762 TI - Quality of life in pregnant women with epilepsy versus women with epilepsy. AB - It is assumed that 25% of patients with epilepsy are women of fertile age and 0.3% to 0.6% of all children are born of mothers with epilepsy. The aim of this study was to evaluate the quality of life on pregnant with epilepsy and compare with non-pregnant women with epilepsy. We evaluated two groups (Experimental Group--29 pregnant women with epilepsy and Control Group--30 women with epilepsy); they were attended at the HC/UNICAMP. The patients had three meetings to carry out and implement the anamnesis and the application of QQV-65. There were no significant differences in the measurement of quality of life when comparing both groups. However, when we analyzed individually in the pre- and post-partum periods, we observed significant differences in health aspects (p=0.0495), physical (p=0.02868) and emotional (p=0.0253) dimensions in QQV-65. This study shows that pregnancy could be interpreted as a stressor. In late pregnancy when this stressor was removed, women with epilepsy had improvement in their quality of life. PMID- 21625763 TI - Depression and anxiety in a community sample with epilepsy in Brazil. AB - OBJECTIVE: To estimate the frequency of depression/anxiety and to establish the social, epilepsy and psychiatric characteristics in individuals with epilepsy. METHOD: A cross-sectional study was employed to evaluate 153 subjects with epilepsy who were identified in a previous community-based survey. First, a structured interview was conducted, followed by a psychiatric evaluation. Subjects with depression were compared to those without, and subjects with anxiety were compared to those without. RESULTS: The prevalence of anxiety and depression was 39.4 and 24.4%, respectively. Both were associated with low schooling (OR 3.8, 95% CI 1.6 to 9.0 and OR 2.8, 95% CI 1.2 to 6.5 for depression and anxiety, respectively), lifetime suicidal thoughts (OR 4.4, 95% CI 1.9 to 10.3 and OR 3.6, 95% CI 1.7 to 7.7) and lifetime suicide attempts (OR 9.3, 95% CI 2.6 to 32.8 and OR 6.9, 95% CI 1.8 to 26.4). CONCLUSION: The high rates of depression and anxiety reinforced the need for recognition and treatment of mental disorders in epilepsy. PMID- 21625764 TI - The role of magnesium sulfate in prevention of seizures induced by pentylenetetrazole in rats. AB - Magnesium sulfate (MgSO4) has been used to prevent seizures in eclampsia. This study examined the central effects of MgSO4 on different types of pentylenetetrazole (PTZ)-induced seizures. Male Wistar rats were submitted to intracerebroventricular (ICV) administration of MgSO4 at different doses followed by intraperitoneal administration of PTZ. The latency to the onset of the first seizure induced by PTZ was significantly increased by ICV administration of MgSO4 at a dose of 100 ug compared to the control treatment. In addition, the total period during which animals presented with seizures was significantly reduced at this dose of MgSO4. Furthermore, the latency to the onset of the first partial complex seizure was significantly increased by the lowest dose of MgSO4. However, a high dose of MgSO4 had no effect or even potentiated the effect of PTZ. These results suggest that, depending on the dose, MgSO4 may be important in prevention of epileptic seizures. PMID- 21625765 TI - Low-level laser therapy, at 830 nm, for pain reduction in experimental model of rats with sciatica. AB - Chronic pain, resulting from nerve compression, is a common clinical presentation. One means of conservative treatment is low-level laser therapy, although controversial. The aim of this study was to evaluate the effects of two doses of low-level laser, at 830 nm, on pain reduction in animals subjected to sciatica. Eighteen rats were used, divided into three groups: GS (n=6), sciatica and simulated treatment; G4J (n=6), sciatica and treatment with 4 J/cm2; and G8J (n=6), sciatica and irradiation with 8 J/cm2. The right sciatic nerve was exposed and compressed using catgut thread. Five days of treatment were started on the third postoperative day. Pain was assessed by means of the paw elevation time during gait: before sciatica, before and after the first and second therapies, and the end of the fifth therapy. Low-level laser was effective in reducing the painful condition. PMID- 21625766 TI - Lipoic acid effects on glutamate and taurine concentrations in rat hippocampus after pilocarpine-induced seizures. AB - Pilocarpine-induced seizures can be mediated by increases in oxidative stress and by cerebral amino acid changes. The present research suggests that antioxidant compounds may afford some level of neuroprotection against the neurotoxicity of seizures in cellular level. The objective of the present study was to evaluate the lipoic acid (LA) effects in glutamate and taurine contents in rat hippocampus after pilocarpine-induced seizures. Wistar rats were treated intraperitoneally (i.p.) with 0.9% saline (Control), pilocarpine (400 mg/kg, Pilocarpine), LA (10 mg/kg, LA), and the association of LA (10 mg/kg) plus pilocarpine (400 mg/kg), that was injected 30 min before of administration of LA (LA plus pilocarpine). Animals were observed during 24 h. The amino acid concentrations were measured using high-performance liquid chromatograph (HPLC). In pilocarpine group, it was observed a significant increase in glutamate content (37%) and a decrease in taurine level (18%) in rat hippocampus, when compared to control group. Antioxidant pretreatment significantly reduced the glutamate level (28%) and augmented taurine content (32%) in rat hippocampus, when compared to pilocarpine group. Our findings strongly support amino acid changes in hippocampus during seizures induced by pilocarpine, and suggest that glutamate-induced brain damage plays a crucial role in pathogenic consequences of seizures, and imply that strong protective effect could be achieved using lipoic acid through the release or decrease in metabolization rate of taurine amino acid during seizures. PMID- 21625767 TI - Effects of eliminating tension by means of epineural stitches: a comparative electrophysiological and histomorphometrical study using different suture techniques in an animal model. AB - Epineural stitches are a means to avoid tension in a nerve suture. We evaluate this technique, relative to interposed grafts and simple neurorraphy, in a rat model. METHOD: Twenty rats were allocated to four groups. For Group 1, sectioning of the sciatic nerve was performed, a segment 4 mm long discarded, and epineural suture with distal anchoring stitches were placed resulting in slight tension neurorraphy. For Group 2, a simple neurorraphy was performed. For Group 3, a 4 mm long graft was employed and Group 4 served as control. Ninety days after, reoperation, latency of motor action potentials recording and axonal counts were performed. Inter-group comparison was done by means of ANOVA and the non parametric Kruskal-Wallis test. RESULTS: The mean motor latency for the simple suture (2.27+/-0.77 ms) was lower than for the other two surgical groups, but lower than among controls (1.69+/-0.56 ms). Similar values were founding in both group 1 (2.66+/-0.71 ms) and group 3 (2.64+/-0.6 ms). When fibers diameters were compared a significant difference was identified between groups 2 and 3 (p=0.048). CONCLUSION: Good results can be obtained when suturing a nerve employ with epineural anchoring stitches. However, more studies are needed before extrapolating results to human nerve sutures. PMID- 21625768 TI - Factors associated with health-related quality of life for adults with stroke sequelae. AB - OBJECTIVE: To assess health-related quality of life (HRQoL) among adults with stroke sequelae and to identify the factors that influenced the domains of HRQoL. METHOD: The subjects were adults with stroke who were admitted to a rehabilitation program between March and August 2008. They were investigated using the following instruments: Medical Outcomes 36-Item Short-Form Health Survey (SF-36), Functional Independence Measure (FIM) and Beck Depression Inventory (BDI). Sixty-four adults of mean age 58.8 years were evaluated. RESULTS: The individuals who had had their injuries for longer times and who were more independent and younger had better HRQoL regarding physical functioning. Social participation was associated with better HRQoL, and there was a difference between groups with and without depressive symptoms. CONCLUSION: HRQoL was compromised primarily with regard to functional characteristics. Moreover, the presence of depressive symptoms contributed towards a decline in HRQoL. Rehabilitation needs to focus on improving functionality and social participation, which may stimulate reductions in depressive symptoms and improvement of HRQoL. PMID- 21625769 TI - Semi-quantitative analysis of the effects of cyclosporine on remyelination following gliotoxic injection in the brainstem. AB - The use of cyclosporine (CsA) has shown to induce an increase in the density of oligodendrocytes near remyelinating areas following the injection of ethidium bromide (EB), a demyelinating agent, in the rat brainstem. This study was designed in order to evaluate if CsA has the capacity of increasing remyelination. In this context, a comparison between the final balance of myelin repair in CsA treated and non-treated rats was assessed using a semi-quantitative method developed for documenting the extent and nature of remyelination in gliotoxic lesions. Wistar rats were submitted to intracisternal injection of 10 microliters of 0.1% EB. Some were treated during 31 days with CsA (group III--10 mg/kg/day by 7 days and, thereafter, 3 times a week, with a minimal interval of 48 hours) by intraperitonial route. Others were not treated with CsA (group I). A control group was planned receiving into the cisterna pontis 10 microliters of 0.9% saline solution and following after that the same CsA administration protocol (group II). Results clearly demonstrate that in vivo administration of CsA after EB-demyelinating lesions stimulated oligodendrocyte remyelination (mean remyelination scores of 3.72+/-0.25 for oligodendrocytes and 1.04+/-0.39 for Schwann cells) compared to non-treated animals (3.13+/-0.71 and 1.31+/-0.62, respectively), although the mechanisms by which this positive CsA effect occurs are unclear. PMID- 21625770 TI - When your child with epilepsy dies suddenly: febrile seizures are part of the process? AB - Febrile seizures (FS) affect almost 2-5% of children and factors related to an increase susceptibility of children to FS may involve an imbalance of inflammatory cytokines and genetic factors. FS had low morbidity, but may be associated with the occurrence of late chronic epilepsy. Here we describe factors related to FS and its possible correlation with SUDEP. PMID- 21625771 TI - The assessment and management of pain in the demented and non-demented elderly patient. AB - Persistent pain is a frequent health problem in the elderly. Its prevalence ranges from 45% to 80%. Chronic diseases, such as depression, cardiovascular disease, cancer and osteoporosis have a higher prevalence in aged individuals and increase the risk of developing chronic pain. The presence of pain is known to be associated with sleep disorders in these patients, as well as functional impairment, decreased sociability and greater use of the health system, with consequent increase in costs. Alzheimer's disease patients seem to have a normal pain discriminative capacity and they may probably have weaker emotional and affective experience of pain when compared to other types of dementia. Many patients have language deficits and thus cannot properly describe its characteristics. In more advanced cases, it becomes even difficult to determine whether pain is present or not. Therefore, the evaluation of these patients should be performed in a systematic way. There are three ways to measure the presence of pain: by direct questioning (self-report), by direct behavioral observation and by interviews with caregivers or informants. In recent years, many pain scales and questionnaires have been published and validated specifically for the elderly population. Some are specific to patients with cognitive decline, allowing pain evaluation to be conducted in a structured and reproducible way. The next step is to determine the type of painful syndrome and discuss the bases of the pharmacological management, the use of multiple medications and the presence of comorbidities demand the use of smaller doses and impose contra-indications against some drug classes. A multiprofessional approach is the rule in the management of these patients. PMID- 21625772 TI - Musical hallucination associated with hearing loss. AB - In spite of the fact that musical hallucination have a significant impact on patients' lives, they have received very little attention of experts. Some researchers agree on a combination of peripheral and central dysfunctions as the mechanism that causes hallucination. The most accepted physiopathology of musical hallucination associated to hearing loss (caused by cochlear lesion, cochlear nerve lesion or by interruption of mesencephalon or pontine auditory information) is the disinhibition of auditory memory circuits due to sensory deprivation. Concerning the cortical area involved in musical hallucination, there is evidence that the excitatory mechanism of the superior temporal gyrus, as in epilepsies, is responsible for musical hallucination. In musical release hallucination there is also activation of the auditory association cortex. Finally, considering the laterality, functional studies with musical perception and imagery in normal individuals showed that songs with words cause bilateral temporal activation and melodies activate only the right lobe. The effect of hearing aids on the improvement of musical hallucination as a result of the hearing loss improvement is well documented. It happens because auditory hallucination may be influenced by the external acoustical environment. Neuroleptics, antidepressants and anticonvulsants have been used in the treatment of musical hallucination. Cases of improvement with the administration of carbamazepine, meclobemide and donepezil were reported, but the results obtained were not consistent. PMID- 21625773 TI - Where was Joseph Babinski born? AB - There is controversy in the neurological literature about where Joseph Babinski was born, including a myth propounded by various important authors that he was born in Lima, Peru. However, according to the most consistent biographical data, he was in fact born in Paris, France, and became a medical celebrity there and in Poland as well as around the world. PMID- 21625774 TI - Fibromuscular dysplasia as a rare cause of severe ischemic stroke in a young man. PMID- 21625775 TI - SCA2 presenting as an ataxia-parkinsonism-motor neuron disease syndrome. PMID- 21625776 TI - Facial grimacing as a clue for the diagnosis of GM1 type 3 gangliosidosis. PMID- 21625777 TI - Huntington's disease presenting as posterior cortical atrophy. PMID- 21625778 TI - Fluorodeoxyglucose positron emission tomography findings in NMDA receptor antibody encephalitis. PMID- 21625779 TI - Myasthenia gravis complicated with cryptococcal meningitis after thymectomy and long-term immunosuppressive therapy. PMID- 21625780 TI - Postpartum angiopathy with reversible cerebral vasoconstriction syndrome (RCVS): imaging in diagnosis and follow-up. PMID- 21625781 TI - Intracerebral amyloidoma: imaging findings might support preoperative diagnosis. PMID- 21625787 TI - [A dietary counseling program and the quality of a prenatal care service]. PMID- 21625788 TI - [Impact of a dietary counseling program on the gain weight speed of pregnant women attended in a primary care service]. AB - PURPOSE: to evaluate the impact of dietary counseling on controlling weight gain in pregnant women, who were served in a public health service facility. METHODS: the study was conducted at a known health unit located in the metropolitan region of the city of Porto Alegre, in Rio Grande do Sul, Brazil. Three hundred and fifteen pregnant women between the 10th and 29th week of gestation were randomized to Control and Intervention Groups. The Intervention Group received dietary counseling according to nutritional status, and pregnant women in the Control Group were instructed to follow the routine of the health service facility. Weight and height were measured, and the body mass index (BMI) was calculated. The pre-gestational nutritional status was determined according to the following BMI criteria: low weight (<18.5 kg/m2), eutrophy (18.5 to 24.9 kg/m2), overweight (25.0 to 29.9 kg/m2), and obesity (>30 kg/m2). The nutritional status during pregnancy was determined according to the BMI curve for gestational age adopted by the Health Ministry of Brazil. Data were analyzed by the relative risk and respective 95% confidence interval, and by the Student's t-test and chi2 test. Statistical significance was set at p<0.05. RESULTS: the assessment of nutritional status before pregnancy showed that 28.0% of the women were overweight and 4.1% were underweight. In the first and last interview during pregnancy, the rates of prevalence of excessive weight were 36.2 and 46.0%, respectively. The intervention proved to be effective in reducing the rate of weekly weight gain of pregnant women with excess weight (342.2 versus 420.2; p=0.015) and the prevalence of clinical complications (9.2 versus 24.85; p<0.001). CONCLUSIONS: dietary counseling was effective in decreasing the weight gain of pregnant women who were overweight and reducing clinical complications, such as gestational diabetes, preeclampsia, infant low weight, and prematurity in the Intervention Group. PMID- 21625789 TI - [Congenital defects and exposure to pesticides in Sao Francisco Valley]. AB - PURPOSE: to evaluate associations between parental exposure to pesticides and births with congenital defects in Sao Francisco Valley, as well as the demographic profile and the defects found. METHODS: in this case-control study, each case (newborns with congenital defects) had two controls (healthy newborns). The subjects were born in the city of Petrolina, in Sao Francisco Valley, in 2009. The sample consisted of 42 cases and 84 controls. Data were gathered by a structured questionnaire adapted from Latin-American Collaborative Study of Congenital Malformations (ECLAMC), with the addition of questions related to exposure to pesticides, analysis of the medical records and contact with the hospital's pediatrician. The chi2 test was performed with a significance level of 5% to identify the variables with the greatest differences between case and control groups. Odds Ratio (OR) for the sample was calculated, as well as the OR obtained by logistic regression analysis, and finally, multivariate logistic regression analysis was performed. RESULTS: there was a greater exposure to pesticides during pregnancy in infants with congenital defects compared to healthy subjects. Increased risk was observed when at least one parent was exposed to pesticides (adjusted OR = 1.3; 95%CI = 0.4 - 3.9). The sociodemographic variables associated with congenital defects were: low school level, low weight, prematurity, young parents, chronic diseases, and physical factors. Multiple malformations and defects of the musculoskeletal and nervous systems were more frequently found. CONCLUSIONS: the present study suggests an association between exposure to pesticides and the occurrence of congenital defects, although the data were not significant. PMID- 21625790 TI - [Ultrasound-guided core needle biopsy for the diagnosis of fibroepithelial breast tumors]. AB - PURPOSE: to evaluate the concordance rate of ultrasound-guided core needle biopsy followed by excisional biopsy in palpable breast lumps, suggestive of fibroepithelial tumors. METHOD: a retrospective study included 70 biopsies with a histological diagnosis of fibroepithelial tumor in 67 out of 531 patients with breast lesions submitted to ultrasound-guided core needle biopsy with a high frequency (7.5 MHz) linear transducer, using an automatic Bard-Magnum gun and a 14-gauge needle. Cases with a diagnosis of fibroepithelial tumor by core needle biopsy or excisional biopsy and with a diagnosis of fibrosclerosis were included in the study. The agreement between the two biopsy methods was assessed using the Kappa coefficient. RESULTS: excisional biopsy revealed 40 cases of fibroadenoma (57.1%), 19 cases of phyllodes tumor (27.2%), and 11 cases of fibrosclerosis (15.7%). The concordance rate for fibroadenoma was substantial (k = 0.68, 95%CI = 0.45 - 0.91), almost perfect for the phyllodes tumor (k = 0.81, 95%CI = 0.57 - 1.0), and moderate for fibrosclerosis (k = 0.58, 95%CI = 0.36 - 0.90). CONCLUSIONS: the core needle biopsy is a minimally invasive method that has "substantial" to "almost perfect" concordance rate with excisional biopsy. Fibrosclerosis should be considered in the differential diagnosis of fibroepithelial tumors. PMID- 21625791 TI - [Elevated blood pressure in women with polycystic ovary syndrome: prevalence and associated risk factors]. AB - PURPOSE: to investigate the prevalence of elevated blood pressure (BP) in patients with polycystic ovary syndrome (PCOS) and to correlate the BP levels with other cardiovascular risk factors. METHODS: a cross-sectional study was conducted on 113 PCOS women (26.2+/-4.3 years) and on a Control Group of 242 healthy women from the general population (26.8+/-5.0 years). The variables considered were: systolic and diastolic BP, anthropometric parameters and plasma levels of glucose, total cholesterol, HDL-cholesterol, and triglycerides. The BP values were classified according to the V Brazilian Guidelines of Hypertension. Statistical analysis was performed by intergroup comparison with the Student's t test and chi2 test, and correlation analysis was performed using Pearson's coefficient. RESULTS: the PCOS Group showed a significantly higher prevalence of altered BP (>130/85 mmHg) than the Control Group (18.6 versus 9.9%, respectively; p<0.05). PCOS women had higher mean systolic BP, body mass index (BMI), waist circumference (WC), triglycerides and fasting glucose, and lower HDL-cholesterol, compared to the Control Group (p<0.01). In the PCOS Group, the values of systolic and diastolic BP showed a significant positive correlation with age, BMI, WC, and triglycerides (p<0.05). CONCLUSIONS: according to the results obtained, it is possible to conclude that the frequency of women with BP values above the normal limit was significantly higher in the PCOS Group than in the Control Group. Additionally, the BP values also correlated with other cardiovascular risk factors. These findings underscore the importance of preventive strategies in PCOS women, in order to prevent pathological events related to the cardiovascular system. PMID- 21625792 TI - [Analysis of codon 72 polymorphism of the TP53 gene in infertile women with and without endometriosis]. AB - PURPOSE: to evaluate the frequency of TP53 codon 72 polymorphism in infertile women with endometriosis, women with idiopathic infertility, controls and its relation to the disease. METHODS: a case-control study that included 198 infertile women with endometriosis, 70 women with idiopathic infertility and 169 fertile women without endometriosis as control. Detection of TP53 codon 72 gene polymorphism (rs1042522, Arg/C:Pro/G), that promotes a C/G exchange in the coding region of the gene, was performed by real time Polymerase Chain Reaction (PCR), using the TaqMan system of primers, that flank the implicated region and probes labeled with different fluorescent dyes, one for allele C and other for allele G. When two dyes were observed, the patient was considered to be heterozygous CG. In the presence of only one dye, the individual was considered to be homozygous CC or GG. The chi2 test was used to compare allele and genotype frequencies between groups. All p-values were two-tailed and a p-value <0.05 was considered to be statistically significant. RESULTS: we found no statistically significant difference in the distribution of TP53 codon 72 polymorphism genotypes CC, CG or GG (p=0.7) and alleles C or G (p=0.4) between infertile patients with endometriosis and controls (p=0.4), regardless of the stage of the disease. In relation to infertility, no statistically significant difference in the genotype or allele distribution (p=1.0 and p=0.9, respectively) was observed between idiopathic infertile women and controls. Considering the dominant inheritance model, again, no statistically significant difference was found even in the endometriosis (p=0.5) or the idiopathic infertility group (p=0.9) when compared to controls. Regarding the recessive inheritance model no statistically significant difference was found, with p=0.6 and p=1.0, respectively, for the endometriosis and idiopathic infertility groups. CONCLUSION: the results suggest that the TP53 codon 72 polymorphism does not confer genetic susceptibility to endometriosis and/or infertility in the Brazilian population, not even the severe form of the disease. PMID- 21625793 TI - [Neoadjuvant chemotherapy followed by radical surgery in pregnant patient with invasive cervical cancer: case report and literature review]. AB - The treatment options for pregnant patients with invasive cervical cancer (ICC) depend on gestational age, clinical stage and the patient's wishes. Some authors have reported cases of neoadjuvant chemotherapy followed by radical surgery in these patients. The aim of this paper was to revisit this subject and to add a new case and review the literature. We report the case of a 30 year-old woman in the 24th week of gestation. She was diagnosed with ICC (squamous cell carcinoma grade 2), stage IIB (International Federation of Gynecology and Obstetrics - FIGO). Nulliparous, the patient refused to interrupt the pregnancy. After meticulous counseling, the patient accepted treatment with neoadjuvant chemotherapy (cisplatin 75 mg/m2 and vincristine 1 mg/m2) and subsequent evaluation of radical surgery concomitant to a cesarean section. Four complete cycles of chemotherapy were administered without delays or significant adverse effects. A few days before the date scheduled for surgery, the patient was admitted in advanced labor (37th week of gestation). Due to tumor clinical response, the obstetric team decided to monitor the labor, and the patient gave birth to a male newborn (2,450 g) uneventfully. Radical surgery was performed three days after birth, and histopathology analysis revealed carcinoma confined to the cervix without lymphatic involvement. Mother and son are in good general condition 12 months after delivery. Cisplatin-based chemotherapy during the second or third trimester of pregnancy appears to be a safe option for patients who do not wish to interrupt a pregnancy while awaiting fetal maturity. However, additional studies are needed to confirm the prognosis and assure the safety of newborns and patients. PMID- 21625794 TI - [Obstetric ultrasound between the 11th and 14th weeks: beyond the screening for chromosomal abnormalities]. AB - This is a traditional (narrative) review with the objective of highlighting the contribution of obstetric ultrasonography (US) between the 11th and 14th week of pregnancy, commonly called first trimester anomaly scan. In addition to being used for the screening of chromosomal anomalies, US can be employed during this period to confirm or determine gestational age, evaluate fetal anatomy, diagnose malformations, screen major structural abnormalities and genetic syndromes, define the prognosis of pregnancy, diagnose and characterize multiple pregnancies, and screen preeclampsia and intrauterine growth restriction. The most important studies about this subject published between 1990 and 2010 in the Cochrane and PubMed libraries were included. The selected studies can be classified with scientific levels I to III. PMID- 21625795 TI - [Impact of a dietary counseling program on the gain weight speed of pregnant women attended in a primary care service]. PMID- 21625797 TI - Assessment of the scientific-technological production in molecular biology in Brazil (1996-2007): the contribution of genomics programs. AB - Several genome sequencing programs were launched in Brazil by the end of the nineties and the early 2000s.The most important initiatives were supported by the ONSA program (http://watson.fapesp.br/onsa/Genoma3.htm) and aimed at gaining domain in genomic technology and bringing molecular biology to the state of art. Two main sets of data were collected in the 1996-2007 period to evaluate the results of these genome programs: the scientific production (Scopus and Web of Science databases) and the register of patents (US Patent and Trademark Office), both related to the progress of molecular biology along this period. In regard to the former, Brazil took a great leap in comparison to 17 other developed and developing countries, being only surpassed by China. As to the register of patents in the area of molecular biology, Brazil's performance lags far behind most of the countries focused in the present study, confirming the Brazilian long standing tendency of poor achievements in technological innovations when compared with scientific production. Possible solutions to surpass this inequality are discussed. PMID- 21625798 TI - Birth weight patterns by gestational age in Brazil. AB - BACKGROUND AND OBJECTIVES: We present an updated birth weight-for-gestational-age portrait, based on nearly 8 million observations of an ethnic-mixed population. It comprises the first comprehensive charts with Brazilian data. This contribution intends to assist clinicians in classifying fetal growth, to provide a reference for investigations of predictors and to show the consequences of small and large patterns for gestational age delivery. Most of the reference data for assessing birth weight for gestational age deal with insufficient sample size, especially at low gestational age. Population-based studies with considerably large sample size refer to data collected more than 15 years ago. METHODS: We accomplished a population-based study on births in all the Brazilian states from 2003 to 2005. Results were based on 7,993,166 singletons. We constructed the 3(rd), 5(th), 10(th), 25(th), 50(th), 90(th), 95(th) and 97(th) smoothed percentiles curves and gender-specific tables from 22 to 43 completed weeks. RESULTS: The resulting tables and graphical representation provide a gender-specific reference to access the birth weights distribution according to the gestational age in the Brazilian population. CONCLUSIONS: This is the first population-based reference constructed on a developing country data. These charts could provide an important tool to improve clinical assessment of growth in newborns. PMID- 21625799 TI - Production, purification and characterization of a thermostable beta-1,3 glucanase produced by Moniliophthora perniciosa. AB - The enzyme glucanase from Moniliophthora perniciosa was produced in liquid medium and purified from the culture supernatant. A multivariate statistical approach (Response Surface Methodology - RSM) was employed to evaluate the effect of variables, including inducer (yeast extract) and fermentation time, on secreted glucanase activities M. perniciosa detected in the culture medium. The crude enzyme present in the supernatant was purified in two steps: precipitation with ammonium sulfate (70%) and gel filtration chromatography on Sephacryl S-200. The best inducer and fermentation time for glucanase activities were 5.9 g L(-1) and 13 days, respectively. The results revealed three different isoforms (GLUI, GLUII and GLUIII) with purification factors of 4.33, 1.86 and 3.03, respectively. The partially purified enzymatic extract showed an optimum pH of 5.0 and an optimum temperature of 40 degrees C. The enzymatic activity increased in the presence of KCl at all concentrations studied. The glucanase activity was highest in the presence of 0.2 M NaCl. The enzyme showed high thermal stability, losing only 10.20% of its specific activity after 40 minutes of incubation at 90 degrees C. A purified enzyme with relatively good thermostability that is stable at low pH might be used in future industrial applications. PMID- 21625801 TI - Growth ring analysis of fossil coniferous woods from early cretaceous of Araripe Basin. AB - Growth ring analysis on silicified coniferous woods from the Missao Velha Formation (Araripe Basin - Brazil) has yielded important information about periodicity of wood production during the Early Cretaceous in the equatorial belt. Despite warm temperatures, dendrological data indicate that the climate was characterized by cyclical alternation of dry and rainy periods influenced by cyclical precipitations, typical of tropical wet and dry or savanna climate. The abundance of false growth rings can be attributed to both occasional droughts and arthropod damage. The present climate data agree with palaeoclimatic models that inferred summer-wet biomes for the Late Jurassic/Early Cretaceous boundary in the southern equatorial belt. PMID- 21625800 TI - Anti-inflammatory, antinociceptive, and antipyretic effects of methanol extract of Cariniana rubra stem bark in animal models. AB - Cariniana rubra Miers (Lecythidaceae), popularly known as "jequitiba-vermelho'', is a large Brazilian tree whose bark is used in infusion and decoction for the treatment of inflammatory conditions. This study aims to assess the anti inflammatory, antinociceptive, and antipyretic effects of Cariniana rubra methanolic stem bark extract (EM Cr) using experimental animals. Anti inflammatory activity of EM Cr was tested on carrageenan and dextran-induced rat paw edema, carrageenan-induced pleurisy in rats and acetic acid-increase vascular permeability in mice. Antinociceptive and antipyretic activities were evaluated using acetic acid-induced writhing, formalin and hot-plate tests in mice, as well as brewer's yeast-induced pyrexia in rats. The extract inhibitied carrageenan and dextran-induced edema, reduced exudate volume and leukocyte migration on the carrageenan-induced pleurisy and on the vascular permeability increase induced by acetic acid. The EM Cr inhibited nociception on the acetic acid-induced writhing and in the second phase of formalin test, and decreased rectal temperature. It was, however, inactive against thermal nociception.Phytochemical analysis with EM Cr showed the occurrence of saponins, triterpenes, sterols and phenolic compounds. Phytosterols (beta-sitosterol, stigmasterol), pentacyclic triterpenes (alpha- and beta-amyrin as a mixture), arjunolic acid, a phytosterol glycoside (sitosterol 3-O-beta-D-glucopyranoside), and triterpenoid saponins (28-beta glucopyranosyl-23-O-acetyl arjunolic acid; 3-O-beta-glucopyranosyl arjunolic acid and 28-O-[alpha-L-Rhamnopyranosyl-(1->2)-beta-glucopyranosyl]-23- O-acetyl arjunolic acid) were the main identified compounds. It can be presumed that EM Cr caused their effects by inhibiting the liberation and/or action of different inflammatory mediators. These findings support the traditional use of Cariniana rubra preparations to treat inflammation. PMID- 21625802 TI - [Associated antimicrobial resistance in Enterococcus spp. clinical isolates]. AB - INTRODUCTION: The increasing prevalence of enterococci strains in hospitals, particularly among isolates of vancomycin-resistant enterococci (VRE), poses important problems because of the limited effect of antimicrobial therapy for enterococcal infections. METHODS: This work presents a retrospective investigation of quantitative in vitro susceptibility data for the range of antimicrobials against Enterococcus spp. isolates and evaluation of the association of resistance between antimicrobial agents recommended as the treatment of choice for infections caused by VRE through calculation of the relative risk. RESULTS: Of the 156 enterococci isolates, 40 (25.6%) were resistant to 3 or more antimicrobials, including 7.7% (n = 12/156) vancomycin resistant. The association of elevated resistance was more pronounced among VRE isolates against alternative and primary antimicrobials for the treatment of infections caused by these pathogens, including ampicillin (100%, RR = 7.2), streptomycin (90.9%, RR = 4.9), rifampin (91.7%, RR = 3.1) and linezolid (50%, RR = 11.5), despite high susceptibility to this drug (94.9%). CONCLUSIONS: The significant associated resistance to alternative and first choice antimicrobials used in the treatment of serious infections of strains with the VRE phenotype and that require a combined therapeutic regime, revealed even more limited therapeutic alternatives in the institution analyzed. PMID- 21625803 TI - [Alternative transmission mechanisms of Trypanosoma cruzi in Brazil and proposals for their prevention]. AB - INTRODUCTION: Following advances in the control of vector and blood transfusion transmission of Chagas disease, alternative mechanisms of transmission have become more relevant. This article discusses the importance of each one of these alternative mechanisms and the measures to prevent them. METHODS: A review was conducted of the scientific literature concerning alternative transmission mechanisms of Trypanosoma cruzi occurring in Brazil and the measures to prevent them. PubMed and BVS databases were consulted. RESULTS: Twenty-five publications describing alternative mechanisms of transmission of Chagas disease were identified. CONCLUSIONS: Oral transmission, through ingestion of contaminated food items has been the most frequent mode of transmission in Brazil in recent years. Other alternative mechanisms of transmission occur less frequently. It is important to understand these occurrences, especially now that vector transmission of the parasite is under control. Preventive measures have been presented, according to each of the situations considered, in line with current knowledge. PMID- 21625804 TI - Comparison of the performance of polymerase chain reaction and pp65 antigenemia for the detection of human cytomegalovirus in immunosuppressed patients. AB - INTRODUCTION: Human cytomegalovirus (HCMV) is often reactive in latently infected immunosuppressed patients. Accordingly, HCMV remains one of the most common infections following solid organ and hemopoietic stem cell transplantations, resulting in significant morbidity, graft loss and occasional mortality. The early diagnosis of HCMV disease is important in immunosuppressed patients, since in these individuals, preemptive treatment is useful. The objective of this study was to compare the performance of the in-house qualitative polymerase chain reaction (PCR) and pp65 antigenemia to HCMV infection in immunosuppressed patients in the Hospital de Clinicas of Porto Alegre (HCPA). METHODS: A total of 216 blood samples collected between August 2006 and January 2007 were investigated. RESULTS: Among the samples analyzed, 81 (37.5%) were HCMV-positive by PCR, while 48 (22.2%) were positive for antigenemia. Considering antigenemia as the gold standard, sensitivity, specificity, positive predictive values and negative predictive values for PCR were 87.5%, 76.8%, 51.8% and 95.5% respectively. CONCLUSIONS: These results demonstrated that qualitative PCR has high sensitivity and negative predictive value (NPV). Consequently PCR is especially indicated for the initial diagnosis of HCMV infection. In the case of preemptive treatment strategy, identification of patients at high-risk for HCMV disease is fundamental and PCR can be useful tool. PMID- 21625805 TI - [Intrauterine growth retardation, low birth weight and prematurity in neonates of pregnant women with malaria in Colombia]. AB - INTRODUCTION: Association between malaria and pregnancy complications, such as prematurity, intrauterine growth restriction, low birthweight and infant mortality has been reported. These effects have been studied widely in areas hyperendemic for malaria, but studies in low-endemic areas are scarce. The study investigated the relation between gestational malaria and low birthweight and intrauterine growth retardation in neonates of a malarial endemic region in Colombia, between 1993 and 2007. METHODS: The pattern of development in 1,716 neonates of women with and without malaria infection during pregnancy was evaluated in a cohort study. A total of 394 infected (27% by P. falciparum and 73% by P. vivax) and 1,322 noninfected pregnant women were followed. RESULTS: Exposure to gestational malaria was associated with increased risk of low birth weight (RR = 1.37; 1.03-1.83), short height (RR = 1.52; 1.25-1.85), intrauterine growth retardation (RR = 1.29; 1.0-1.66) and prematurity (RR = 1.68; 1.3-2.17). Prematurity was 77% higher in infants of mothers with malaria by P. falciparum than infants of mothers with malaria by P. vivax (RR = 1.77; 1.2-2.6). CONCLUSIONS: Low birth weight and intrauterine growth retardation were associated with malaria during pregnancy. Infection with P. vivax was related with adverse effects on the newborn, similar to that reported for P. falciparum. PMID- 21625806 TI - [Risks of transmitting rabies virus from captive domiciliary common marmoset (Callithrix jacchus) to human beings, in the metropolitan region of Fortaleza, state of Ceara, Brazil]. AB - INTRODUCTION: In the State of Ceara, a new variant of the rabies virus was identified associated with cases of human rabies transmitted by common marmosets (Callithrix jacchus), which are frequently kept as pets. This new variant does not present antigenic proximity or genetic relationship to variants of the virus isolated from bats and terrestrial mammals from the American continent. The present study aimed to evaluate the risk factors of rabies virus transmission from common marmosets (C. jacchus) maintained as pets in the metropolitan region of Fortaleza, State of Ceara, Brazil, to human beings. METHODS: A questionnaire focusing on animal management and interaction between humans and primates was applied to individuals who had marmosets in the municipalities of Aquiraz and Maranguape. In order to evaluate the presence of rabies antigens by direct immunofluorescence test (DIF), samples of saliva were collected from domiciliary captive marmosets. Based on the detection of rabies antigens, biopsy samples of central nervous system (CNS) were analyzed. RESULTS: Analysis of questionnaire data verified that a close relation exists between humans and their pet marmosets, especially during management practices. Additionally, these people showed minimal knowledge regarding rabies, which represents a greater risk of infection. Of the 29 saliva samples evaluated, one (3.4%) was positive for DIF reaction and of the 11 CNS samples, three (27.3%) were positive. CONCLUSIONS: Laboratory data are in agreement with the questionnaire findings, which confirm an increased risk of rabies virus transmission due to the close relation between humans and marmosets. PMID- 21625808 TI - Considerations about the 2011 consensus of the Brazilian Society of Rheumatology for diagnosis and early assessment of rheumatoid arthritis. PMID- 21625809 TI - 2011 Consensus of the Brazilian Society of Rheumatology for diagnosis and early assessment of rheumatoid arthritis. AB - OBJECTIVE: Develop guidelines for management of rheumatoid arthritis (RA) in Brazil, focusing on diagnosis and early assessment of the disease. METHOD: Literature review and expert opinions of RA Committee members of the Brazilian Society of Rheumatology. RESULTS AND CONCLUSIONS: The following ten reccommendations were established: 1) RA diagnosis should be established considering clinical findings and complementary test results; 2) Special attention should be given to the differential diagnosis of arthritis; 3) Rheumatoid factor (RF) is an important diagnostic test, but has limited sensitivity and specificity, mainly in early RA; 4) Anti-CCP (anti-cyclic citrullinated peptide antibody) is a marker with sensitivity similar to that of the RF, but with higher specificity, mainly in the initial phase of disease; 5) Although unspecific, acute-phase reactants should be measured in patients with clinical suspicion of RA; 6) Conventional radiography should be performed for diagnostic and prognostic assessment of the disease. When necessary and available, ultrasound and magnetic resonance may be used; 7) Rheumatoid arthritis classification criteria (ACR/EULAR 2010), although not yet validated, may be used as a guide to aid in diagnosing patients with early RA; 8) One of the combined disease activity indices should be used to assess disease activity; 9) At least one of the functional capacity assessment instruments, such as mHAQ or HAQ-DI, should be regularly used; 10) At the early assessment of the disease, the presence of worse prognostic factors, such as polyarticular involvement, high titers of RF and/or anti-CCP, and early joint erosion, should be investigated. PMID- 21625810 TI - Use of biological therapies in rheumatoid arthritis management: a comparison between the main worldwide and brazilian recommendations. AB - INTRODUCTION: Rheumatoid arthritis (RA) is a chronic systemic and progressive inflammatory disorder of the joints, which can result in deformity and functional disability. The diagnosis, treatment, and follow-up of patients with RA vary worldwide. The major societies of rheumatology, as well as governmental agencies in most countries, have tried to establish recommendations addressing diagnosis, treatment, and follow-up of RA. Despite the rapid advance in discovering new drugs, with increasingly efficient therapeutic responses, these recommendations have not been updated accordingly. Thus, efforts should be focused on standardizing the procedures established. OBJECTIVE: Compare the main international recommendations for treatment of RA with the Brazilian protocols of the Brazilian Society of Rheumatology and Ministry of Health. METHODS: The protocols of the following entities for treating RA were assessed: Brazilian Ministry of Health, Brazilian Society of Rheumatology, PANLAR/GLADAR, American College of Rheumatology, European League Against Rheumatism (EULAR), and Mexican College of Rheumatology. RESULTS: Significant differences were identified between the several recommendations, especially regarding the criteria for beginning biological therapies, hierarchic sequence for using available biological drugs, and for suspending or switching them. CONCLUSIONS: The recommendations for treatment of RA should be more frequently updated. The worldwide standardization of criteria for elaborating recommendations would be of great value to provide similar guidance to rheumatologists in countries and regions throughout the world. PMID- 21625811 TI - Incidence and clinical-laboratory aspects of systemic lupus erythematosus in a Southern brazilian city. AB - INTRODUCTION: Brazilian epidemiological studies on systemic lupus erythematosus (SLE) are scarce, and currently available data originate almost entirely from international literature. OBJECTIVES: To determine the incidence and some clinical and laboratory characteristics of patients with SLE in the municipality of Cascavel, state of Parana, Brazil. PATIENTS AND METHODS: Data were collected from August 2007 to July 2008 in all health services of Cascavel providing health care in rheumatology: a university-affiliated hospital, a public outpatient clinic, and three private clinics. RESULTS: The study identified 14 patients diagnosed with SLE, which resulted in an estimated incidence of 4.8 cases/100,000 inhabitants/year. All patients were female, and the mean age was 41.5 years. The highest incidence of disease occurred between 30 and 39 years of age, and 92.8% of patients met at least four of the 11 American College of Rheumatology (ACR) criteria for diagnosis of SLE. The drug treatment of patients was also assessed and proved to be in accordance with the Brazilian Consensus for Treatment of SLE. CONCLUSION: The incidence obtained in the municipality of Cascavel is close to those reported in international studies. PMID- 21625812 TI - Musculoskeletal system assessment in an emergency room. AB - OBJECTIVE: Musculoskeletal conditions have an enormous and growing impact worldwide. In spite of that, some clinicians are not confident in their own musculoskeletal examination skills. This study aimed to evaluate the prevalence of musculoskeletal symptoms in an emergency room, and the frequency of musculoskeletal physical examination description on those cases. METHODS: This was a cross-sectional study. We performed a systematic analysis of medical files at the emergency room of the University Hospital of the Federal University of Santa Catarina, Brazil, from April 24th to 30th, 2009. RESULTS: We analyzed 392 files, where 41.5% of patients were male and mean age was 38.7 +/- 17.2 years old. Sixty nine out of 392 patients (17.6%) presented with a musculoskeletal complaint. The most common musculoskeletal complaint was low back pain (33/69). Only 49.2% of patients with a musculoskeletal chief complaint had a specific physical examination registered on the files. Patients with musculoskeletal complaints had lower registrations of abdominal examination (46% versus 62%, P = 0.01) and vital signs (46% versus 66%, P = 0.002), but a higher frequency of musculoskeletal examination registration (49% versus 0.6%, P = 0.00). CONCLUSIONS: Our study confirms other observations worldwide. Musculoskeletal complaints are frequent in a emergency room setting and in spite of that it is suggested that musculoskeletal symptoms are poorly evaluated, which is probably related to an insufficient musculoskeletal education. It is essential that medical schools place more emphasis on these conditions so that young physicians will be more prepared to deal with these common diseases. PMID- 21625813 TI - Swimming training and functional assessment of sciatic nociception in rats. AB - INTRODUCTION: The protective effect of exercise on individuals with peripheral neuropathy is controversial. OBJECTIVE: To assess the influence of physical training of moderate and progressive intensity on nociception of rats in an experimental sciatica model. MATERIALS AND METHODS: The study assessed 18 rats divided into the following three groups: control group (CG); low-intensity swimming group (LISG); progressive-time swimming group (PTSG). To meet the physical training protocol, the animals were placed in a water tank for six weeks, three alternate days per week. In LISG group, the animals swam ten minutes per session, and, in PTSG group, the animals began swimming for ten minutes, and had a ten-minute increase per week. In CG group, the animals swam less than one minute per day, only to get used to the water environment. After finishing the swimming training, the experimental sciatica model protocol was started with constrictive ligature of the sciatic nerve with chromic catgut. Nociception was assessed using the functional disability test, which measures, in a one-minute interval, the time during which the animal holds its hind paw (THHP) in a guarded position. Assessments were performed preoperatively, and on the third, sixth, and tenth post-operative (PO) days. RESULTS: All groups showed a significant increase in THHP on third, sixth, and tenth PO days, as compared to preoperative values. In CG, no significant change was observed in PO assessments, unlike what happened in LISG and PTSG groups. CONCLUSION: Exercise produced greater hyperalgesia. PMID- 21625814 TI - Rheumatoid arthritis and metabolic syndrome. AB - In the past 20 years, the life expectancy of patients with rheumatoid arthritis (RA) has been shown to be reduced by three to ten years as compared to that of the general population. Currently, cardiovascular disease (CVD) is the major cause of death in patients with RA, and acute myocardial infarction can be up to four times more frequent in these patients. The autoimmune systemic inflammatory response, along with the presence of metabolic syndrome (MetS), doubles the risk for fatal or non-fatal CVD and coronary atherosclerosis, regardless of age and sex. Rheumatoid arthritis has been associated with increased prevalence of MetS, but its role in the different characteristics of the disease, such as disease duration, activity, and treatment with glucocorticoids, is not well defined. This study aimed at reviewing the prevalence of MetS and the factors implicated in the development of atherosclerosis in RA patients, assessing the clinical aspects of RA and its association with the development of MetS. PMID- 21625815 TI - Non-pharmacological therapy and complementary and alternative medicine in fibromyalgia. AB - Fibromyalgia is a chronic painful syndrome that affects up to 5% of the world population. It is associated with sleep and mood disorders, fatigue, and functional disability. Its pathogenesis involves a disorder of the central modulation of pain, impairment of the descending inhibitory system, and hyperactivity of substance P. Because of the extensive symptomatology of patients with fibromyalgia and its multifactorial pathogenesis, its ideal treatment requires a multidisciplinary approach including the association of pharmacological and non-pharmacological therapies. The pharmacological therapy currently recommended for the syndrome includes antidepressants, calcium-channel modulators, muscle relaxants, and analgesics. In most cases, the non pharmacological treatment consists of patient education, supervised aerobic physical activity, and cognitive-behavioral therapy. However, many patients do not respond satisfactorily, or have side effects associated with the long-term use of drugs, in addition to reporting difficulties in adhering to a therapy based on exercises and physical medicine. Thus, physicians and patients are increasingly interested in an alternative and complementary therapy for fibromyalgia. This review approaches the different therapeutic modalities used in fibromyalgia, emphasizing the evidence of non-pharmacological therapy and use of alternative and complementary medicine for these patients. PMID- 21625816 TI - Posterior ankle impingement syndrome: a diagnosis rheumatologists should not forget. Two case reports. AB - The ankle is a common site of painful symptoms in athletes and nonathletes. Posterior ankle pain can be the end result of several pathologies, and a diagnostic challenge for rheumatologists. The posterior ankle impingement syndrome, also known as os trigonum syndrome and posterior tibiotalar compression syndrome, is a clinical disorder characterized by acute or chronic posterior ankle pain triggered by forced plantar flexion, which causes chronic repetitive microtrauma. Pathology of the os trigonum-talar process is the most common cause of this syndrome, but there are other causes, such as tenosynovitis of the flexor hallucis longus, ankle osteochondritis, subtalar joint disease, and fracture. Diagnosis is based on clinical history and physical examination, and complemented by findings on plain radiography (RX), ultrasound (US), scintigraphy, computed tomography (CT), and magnetic resonance imaging (MRI). It is worth noting that RX has low cost and good sensitivity, US can provide guidance to therapeutic infiltrations, and MRI allows the assessment of surrounding soft tissues. PMID- 21625817 TI - Systemic Lupus Erythematosus with muscle weakness due to Myasthenia Gravis. AB - Systemic lupus erythematosus (SLE) and myasthenia gravis (MG) are autoimmune diseases, whose association in the same patient is rarely reported. Both pathologies share the following characteristics: affect mainly young women; alternate exacerbation and remission periods; and have positive antinuclear antibody (ANA) test. This case report assesses possible diagnostic hypotheses for the clinical findings of eyelid ptosis and proximal muscle weakness in a female patient recently diagnosed with SLE, who evolved with associated MG. PMID- 21625818 TI - Associated factors to total cholesterol: school based study in southern Brazil. AB - BACKGROUND: Evidence has suggested that a significant proportion of children and adolescents has high levels of total cholesterol. OBJECTIVE: To estimate the prevalence of hypercholesterolemia and associated factors in 07-12 year-old school children. METHODS: School based cross sectional study of a random sample of 1,294 07-12 year-old school children from Caxias do Sul/RS, Brazil. The students answered an interview with information on socioeconomic status, food habits, and physical and leisure activities. Total cholesterol, cardiorespiratory fitness, body mass and height were measured to calculate body mass index. For the data treatment, univariate, bivariate, and multivariate analyzes were used. RESULTS: The multivariate analysis identified that individuals from high socioeconomic level (OR: 1.70; CI: 1.05-2.75), of female gender (OR: 1.32; CI: 1.03-1.67), and overweight (OR: 1.40; IC: 1.10-1.77) had increased chances of having increased total cholesterol (> 3rd tercile). CONCLUSION: High total cholesterol levels on 07-12 year-old school children are associated with high socioeconomic level, female gender, and overweight. Encouraging an active life style and appropriate dietary habits can help control cholesterol levels and reduce risk factors. PMID- 21625819 TI - Pulse wave velocity in young adults: study of Rio de Janeiro. AB - BACKGROUND: Pulse wave velocity (PWV) can be a marker of cardiovascular impairment, but there are few studies in young adults. OBJECTIVE: To evaluate the association between blood pressure (BP), current anthropometric and metabolic variables and those obtained 13 years earlier, in childhood and adolescence, with PWV. METHODS: Sixty individuals were followed longitudinally and split into two groups according to the percentile of blood pressure (BP) obtained 13 years earlier: Group 1 (G1): BP percentile < 50 (n = 25, 11M, 26.4 years old) and Group 2 (G2): BP > 95 percentile (n = 35, 19M, 25.4 years old). The individuals underwent clinical evaluation, laboratory analysis and measurements of PWV through the Complior method. RESULTS: G1 showed higher mean age; G2 showed greater mean weight, systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), PWV and blood glucose, and lower mean HDL cholesterol. SBP, MAP and heart rate (HR) obtained during childhood and adolescence significantly correlated with PWV. Current weight, height, waist-hip ratio, SBP, DBP, pulse pressure (PP), MAP and creatinine presented a positive and significant correlation with PWV. A comparison of the average PWV adjusted for SBP, DBP, SBP and DBP, MAP and PP showed no statistically significant difference between groups. CONCLUSION: The percentile of BP in childhood/adolescence related to arterial distensibility assessed by PWV 13 years later. Changes in PWV can be identified in young individuals suggesting that early vascular impairment may be present in this age group, also related to blood pressure, anthropometric and metabolic variables. PMID- 21625820 TI - Upregulation of mRNA myocardium calcium handling in rats submitted to exercise and food restriction. AB - BACKGROUND: Chronic exercise and food restriction (FR) have directionally opposite changes in transcription of molecular structures of calcium handling and thyroid hormone (TH) status. OBJECTIVE: Evaluate the association of chronic exercise and FR on serum thyroid hormones and gene transcription of molecular structures of intracellular calcium transients and thyroid receptors in myocardium of rats. METHODS: Male Wistar Kyoto rats, divided into two groups: control (C, n = 7), FR (R50, n = 7), chronic exercise (EX, n = 7) and chronic exercise + FR (EX50, n = 7). FR was of 50% and exercise was swimming (1 hour/day, 5 days/week, during 12 weeks). Serum concentrations of T3, T4 and TSH were determined. The mRNA gene expression of the sarcoplasmatic reticulum calcium pump (SERCA2a), phospholamban (PLB), Na+/Ca+2 exchanger (NCX), calcium channel L-type (L-channel), ryanodine (RYR), calsequestrin (CQS) and HT receptor (TRalpha1 and TRbeta1) of the myocardium was performed by PCR real-time. RESULTS: FR reduced serum levels of T4 and TSH and TRalpha1 mRNA and increased the expression of PLB, NCX and L-channel. Exercise increased the TRbeta1 receptor, L-channel and NCX. The association of exercise and FR reduced plasma T4 and TSH, TRbeta1 mRNA increase, SERCA2a, NCX and PLB, and there was a significant correlation of TRbeta1 with CQS and NXC. CONCLUSION: Chronic exercise and food restriction increased the mRNA of transient Ca2+ proteins; however, TH-receptor axis cannot participate in the transcription of mRNA of myocardial calcium transient proteins. PMID- 21625821 TI - Aging alters the production of iNOS, arginase and cytokines in murine macrophages. AB - The limited amount of information on the primary age-related deficiencies in the innate immune system led us to study the production of inducible nitric oxide synthase (iNOS), arginase, and cytokines in macrophages of young (8 weeks old) and old (72 weeks old) female BALB/c mice. We first evaluated iNOS and arginase inducers on peritoneal (PMPhi) and bone marrow-derived (BMMPhi) macrophages of young BALB/c and C57BL/6 mice, and then investigated their effects on macrophages of old mice. Upon stimulation with lipopolysaccharide (LPS), resident and thioglycolate-elicited PMPhi from young mice presented higher iNOS activity than those from old mice (54.4%). However, LPS-stimulated BMMPhi from old mice showed the highest NO levels (50.1%). Identical NO levels were produced by PMPhi and BMMPhi of both young and old mice stimulated with interferon-gamma. Arginase activity was higher in resident and elicited PMPhi of young mice stimulated with LPS (48.8 and 32.7%, respectively) and in resident PMPhi stimulated with interleukin (IL)-4 (64%). BMMPhi of old mice, however, showed higher arginase activity after treatment with IL-4 (46.5%). In response to LPS, PMPhi from old mice showed the highest levels of IL-1alpha (772.3 +/- 51.9 pg/mL), whereas, those from young mice produced the highest amounts of tumor necrosis factor (TNF) alpha (937.2 +/- 132.1 pg/mL). Only TNF-alpha was expressed in LPS-treated BMMPhi, and cells from old mice showed the highest levels of this cytokine (994.1 +/- 49.42 pg/mL). Overall, these results suggest that macrophages from young and old mice respond differently to inflammatory stimuli, depending on the source and maturity of the cell donors. PMID- 21625822 TI - Immunological and biochemical parameters of patients with metabolic syndrome and the participation of oxidative and nitroactive stress. AB - Metabolic syndrome (MS) is a multifactorial disease involving inflammatory activity and endothelial dysfunction. The aim of the present study was to evaluate the relationship between the changes in lipoperoxidation, in immunological and biochemical parameters and nitric oxide metabolite (NOx) levels in MS patients. Fifty patients with MS (4 males/46 females) and 50 controls (3 males/47 females) were studied. Compared to control (Mann-Whitney test), MS patients presented higher serum levels (P < 0.05) of fibrinogen: 314 (185-489) vs 262 (188-314) mg/dL, C-reactive protein (CRP): 7.80 (1.10-46.50) vs 0.70 (0.16 5.20) mg/dL, interleukin-6: 3.96 (3.04-28.18) vs 3.33 (2.55-9.63) pg/mL, uric acid: 5.45 (3.15-9.65) vs 3.81 (2.70-5.90) mg/dL, and hydroperoxides: 20,689 (19,076-67,182) vs 18,636 (15,926-19,731) cpm. In contrast, they presented lower (P < 0.05) adiponectin: 7.11 (3.19-18.22) vs 12.31 (9.11-27.27) ug/mL, and NOx levels: 5.69 (2.36-8.18) vs 6.72 (5.14-12.43) uM. NOx was inversely associated (Spearman's rank correlation) with body mass index (r = -0.2858, P = 0.0191), insulin resistance determined by the homeostasis model assessment (r = -0.2530, P = 0.0315), CRP (r = -0.2843, P = 0.0171) and fibrinogen (r = -0.2464, P = 0.0413), and positively correlated with hydroperoxides (r = 0.2506, P = 0.0408). In conclusion, NOx levels are associated with obesity, insulin resistance, oxidative stress, and inflammatory markers. The high uric acid levels together with reactive oxygen species generation may be responsible for the reduced NO levels, which in turn lead to endothelial dysfunction. The elevated plasma chemiluminescence reflecting both increased plasma oxidation and reduced antioxidant capacity may play a role in the MS mechanism. PMID- 21625823 TI - Molecular and cellular pathogenesis of autosomal dominant polycystic kidney disease. AB - Autosomal dominant polycystic kidney disease (ADPKD) is one of the most common human life-threatening monogenic disorders. The disease is characterized by bilateral, progressive renal cystogenesis and cyst and kidney enlargement, often leading to end-stage renal disease, and may include extrarenal manifestations. ADPKD is caused by mutation in one of two genes, PKD1 and PKD2, which encode polycystin-1 (PC1) and polycystin-2 (PC2), respectively. PC2 is a non-selective cation channel permeable to Ca(2+), while PC1 is thought to function as a membrane receptor. The cyst cell phenotype includes increased proliferation and apoptosis, dedifferentiation, defective planar polarity, and a secretory pattern associated with extracellular matrix remodeling. The two-hit model for cyst formation has been recently extended by the demonstration that early gene inactivation leads to rapid and diffuse development of renal cysts, while inactivation in adult life is followed by focal and late cyst formation. Renal ischemia/reperfusion, however, can function as a third hit, triggering rapid cyst development in kidneys with Pkd1 inactivation induced in adult life. The PC1-PC2 complex behaves as a sensor in the primary cilium, mediating signal transduction via Ca(2+) signaling. The intracellular Ca(2+) homeostasis is impaired in ADPKD, being apparently responsible for the cAMP accumulation and abnormal cell proliferative response to cAMP. Activated mammalian target for rapamycin (mTOR) and cell cycle dysregulation are also significant features of PKD. Based on the identification of pathways altered in PKD, a large number of preclinical studies have been performed and are underway, providing a basis for clinical trials in ADPKD and helping the design of future trials. PMID- 21625824 TI - Impact of dermoscopy on the management of high-risk patients from melanoma families: a prospective study. AB - Few studies have investigated the impact of dermoscopy on the management of relatives from melanoma families. The objective of this study was to assess the impact of dermoscopy on clinical diagnosis and management decisions in high-risk familial melanoma patients. In a prospective study 132 consecutive patients were recruited from the pigmented lesions clinic of a tertiary reference centre for familial melanoma. Dermatologists expert in dermoscopy identified 49 suspicious pigmented lesions and recorded pre- and post-dermoscopy diagnoses and management decisions. Dermoscopy was performed in 37% of the patients. Two melanomas were identified. Dermoscopy did not influence sensitivity (1.0), but resulted in 42% fewer excisions, increasing specificity from 0.53 to 0.74 (p = 0.031). Dermoscopy resulted in a large reduction in the number of unnecessary excisions. These results suggest that the main effect of dermoscopy in clinical practice for this high risk population is a significant increase in specificity, rather than sensitivity. PMID- 21625825 TI - Food for Thought ... on mapping the human toxome. AB - The report by the National Research Council of the US National Academy of Sciences, Toxicity Testing in the 21st Century: A Vision and a Strategy, has prompted a discussion about renewing regulatory toxicology - especially for chemicals - by harnessing in vitro tests, in silico approaches, and testing in lower organisms. The key change is basing the assessments on mechanisms and toxicant modes of action. Identifying pathways of toxicity (PoT), especially on a larger scale, evidently requires omics technologies. When the PoT is known, a test battery allowing higher throughput than the current approach can be constructed. Here, we propose an extension of this concept to mapping the entirety of PoT in humans: the human toxome. Mapping the human toxome will allow us, for the first time, to conclusively identify substances as nontoxic or to identify nontoxic concentrations of substances (i.e., concentrations at which no relevant PoT are triggered). The concept is explained, and opportunities and obstacles are discussed, aiming to promote an initiative which will form the core of a Human Toxicology Project to implement Toxicology for the 21st Century. PMID- 21625826 TI - Examining the regulatory value of multi-route mammalian acute systemic toxicity studies. AB - Regulatory information requirements for pesticides call for submission of acute systemic toxicity data for up to three different exposure routes (oral, dermal, inhalation) for both active ingredients and formulated products. Similar multi route testing is required in the European Union and elsewhere for industrial chemicals. To determine the value of acute toxicity testing by more than one route, oral-dermal and oral-inhalation concordances among regulatory classifications were examined for large data sets of chemicals and pesticide active ingredients. Across all sectors examined, oral acute toxicity classifications for pure active substances were more severe than those derived from dermal data in more than 98% of cases, which calls into question the value of routine dermal route testing for acute toxicity. Oral classifications were equivalent to or more severe than for the inhalation route for 83% of industrial chemicals and for 48% of pesticides examined. PMID- 21625827 TI - Systematic review of chimpanzee use in monoclonal antibody research and drug development: 1981-2010. AB - This survey examines the extent to which live chimpanzees have been used in monoclonal antibody (mAb) research and the drug approval process. The survey covers 193 scientific articles published during the years 1981-2010, as well as preclinical studies leading to the approval of mAb drugs by the Food and Drug Administration of the United States. The frequency of the articles has decreased by more than two- thirds from their highs in the late 1980's, and the aggregate number of chimpanzees used in these studies has decreased by more than 90%. The experimental protocols ranged from single or multiple blood draws to extraction of body fluids and tissue samples, and to multiple, repeated organ biopsies. Many studies involved infecting the chimpanzee(s) with pathogenic organisms and immunization and infusion protocols. Addressing the health history and status of the chimpanzees was an exception rather than the rule, and anesthesia and analgesia were mentioned only in a small minority of the surveyed articles. In the past two decades, the FDA has approved 32 mAb drugs, but only three of those drugs could be determined to involve the chimpanzee in the preclinical stage. Two of those three drugs have been withdrawn from the market due to their severe adverse effects in human patients. Available alternatives, together with ethical and economic reasons, suggest that the use of the chimpanzee in this manner may not be necessary or appropriate. PMID- 21625828 TI - Local lymph node assay: how testing laboratories apply OECD TG 429 for REACH purposes. AB - The Local Lymph Node Assay (LLNA) is the official method for assessing the allergic contact dermatitis potential of chemicals for the purposes of REACH regulation. The LLNA went through a validation process that allowed the delineation of a robust protocol for performing new tests. The OECD accepted this method in 2002 and published OECD TG 429. The European Chemical Agency (ECHA) recently published data that were submitted in the registration dossiers of chemicals. This database was analysed to determine how testing laboratories apply OECD TG 429. This analysis comes after a detailed analysis of four full study reports that were also prepared for REACH purposes. Although the majority of the tests are fully compliant with OECD TG 429, some showed major deviations, and a number of others used more animals than necessary. This suggests that in vivo tests need to be planned more carefully and consciously to obtain meaningful results with the minimum animal number necessary. PMID- 21625829 TI - A critique of the EC's expert (draft) reports on the status of alternatives for cosmetics testing to meet the 2013 deadline. AB - The 7th Amendment to the EU's Cosmetic Directive (now recast as Regulation 1223/2009) bans the testing of cosmetic ingredients and products on animals, effective 2009. An extension until 2013 was granted, for marketing purposes only, for three endpoints: repeated dose, toxicokinetics, and reproductive toxicity. If the European Commission determines that alternatives for these endpoints are not likely to be available, it can propose a further extension. To this end, the Commission has instructed experts to produce reports on the status of alternatives for the 2013 deadline. We criticized the draft reports on a number of issues. First, the experts fell into the "high fidelity fallacy trap," i.e. asserting that full replication of the in vivo response, as opposed to high predictivity, is required before an animal test can be considered useful for regulatory purposes. Second, the experts' reports were incomplete, omitting various methods and failing to provide data on the validity, reliability, and applicability of all the methods discussed, regardless of whether the methods were in vivo, in vitro, or in silico. In this paper we provide a summary of our criticisms and provide some of the missing data in an alternative proposal for replacement of animal tests by 2013. It is our belief that use of the Threshold of Toxicological Concern (TTC) will be a useful method to mitigate much animal testing. Alternative approaches for carcinogenicity and skin sensitization could be considered sufficient in the very near future, even though these tests are not listed under the 2013 extension. For repeated dose, toxicokinetics, and reproductive toxicity a combination of in vitro methods may be able to provide appropriate protection for consumers, especially when viewed in the context of the poor predictivity of the animal models they replace. We hope the revised report will incorporate these comments, since a more thorough and positive review is required if the elimination of animal testing for cosmetics in Europe and beyond is to be achieved. PMID- 21625830 TI - Joint line is well restored when navigation surgery is performed for total knee arthroplasty. AB - PURPOSE: The incorrect restoration of the joint line during TKA can result in joint instability, anterior knee pain, limited range of motion, and joint stiffness. The joint line level is usually measured only on pre- and post operative radiographs. Current knee navigation systems can now potentially support intra-operatively joint line restoration by controlling the exact amount of the bone-cartilage removed and the corresponding overall thickness of the components implanted. The aim of this study was to assess how well the joint line level is restored and the tibiofemoral overstuffing prevented when standard knee surgical navigation is used carefully also with these purposes. Intra-operative measurements during navigated TKA were taken. METHODS: Sixty-seven primary TKAs were followed prospectively. The variation before and after prosthesis component implantation of the joint line level, both in the femoral and tibial reference, was measured intra-operatively by an instrumented probe. Overstuffing was measured as the difference between the overall craniocaudal thickness of the femoral and tibial prosthesis components inserted and the thickness of the bone cartilage removed. RESULTS: A significant elevation in the joint line level after prosthesis implantation was found with respect to the tibial reference (1.9 +/- 2.4 mm, mean +/- SD), very little to the femoral reference (0.3 +/- 2.1 mm), perhaps accounted for the femur-first operative technique utilized. Overstuffing was on the average of 2.2 +/- 3.0 mm. CONCLUSIONS: These results suggest that a knee navigation system can also support well a proper restoration of the joint line level and limit the risk of overstuffing when relevant measurements are taken carefully during operation. LEVEL OF EVIDENCE: III. PMID- 21625831 TI - Approach to a child with lower airway obstruction and bronchiolitis. AB - Lower airway obstruction can occur at the level of trachea, bronchi or bronchioles. It is characterized clinically by wheeze and hyperinflated chest, apart from other signs of respiratory distress. Common causes include bronchiolitis, asthma, pneumonia, laryngotracheo-bronchitis, congenital malformations and foreign body inhalation. Bronchiolitis usually occurs in children aged 2 months to 2 years. It is most commonly caused by respiratory syncytial virus infection. The diagnosis is mainly clinical, and investigations have a very limited role. Humidified oxygen and supportive therapy are the mainstays of treatment. A trial of inhaled epinephrine or parenteral steroids may be considered for non-responders. It is usually associated with good outcome. PMID- 21625832 TI - Clinical Pearls in pediatric infections. AB - This series of Clinical Pearls presents four cases presenting with infection. Each of these cases had clinical clues to the correct diagnosis, which could be picked up on meticulous history, clinical examination, or basic laboratory investigations. The authors highlight the important lessons to be learnt from each case. The first is a 7 year old boy with recurrent respiratory tract infections since early life. Clinical examination revealed the presence of dextrocardia and situs inversus and bronchiectasis leading to a diagnosis of Primary Ciliary Dyskinesia. The second case is a 1.5-month-old infant who presented with meningitis and increasing head size since birth. CSF examination and CT scanning led to the correct diagnosis of congenital Toxoplasmosis. The next case is an infant with high grade fever and neck swelling. He had the rare Lemierre's syndrome comprising of oro-pharyngeal infection, suppurative thrompbophlebitis of the internal jugular vein and systemic dissemination of septic emboli. The fourth case is a 2-year-old infant with recurrent respiratory tract infections and discharging neck swellings from early life. Repeated testing for tuberculosis was negative. The diagnosis was Chronic granulomatous disease. The authors describe the clinical approach and investigations in these cases; along with an outline of the management. PMID- 21625833 TI - Successful treatment of neonatal chylothorax with octreotide. AB - Chylothorax is a relatively uncommon, but a common form of pleural effusion in the neonates. It may be either congenital or acquired. The efficacy of octreotide therapy for chylothorax is controversial. Herein the authors report successful suppression of chylothorax by octreotide in a newborn who had undergone thoracostomy tube. PMID- 21625834 TI - Juvenile idiopathic arthritis-its rare presentation. AB - The authors report a case of systemic onset juvenile idiopathic arthritis who presented with fever, excruciating neck pain and stiffness followed by respiratory difficulty after few days of onset of illness. The unusual features of the case were: cervical spondylitis without any peripheral joint involvement, myocarditis leading to congestive cardiac failure, though pericarditis being the common cardiovascular manifestation of SOJIA was also associated. PMID- 21625835 TI - Prenatal and perinatal burden on dentoalvelor development: awareness of need for especial dentistry follow up program for premature neonates. PMID- 21625836 TI - Presence of severe iodine deficiency in areas with adequate salt iodization. PMID- 21625837 TI - Assessment of glucose tolerance in multi-transfused adolescent and young adult patients with beta thalassemia. PMID- 21625838 TI - Evaluation of pulmonary hypertension in a child: role of computed tomography. AB - Unexplained pulmonary hypertension in pediatric patient is a diagnostic challenge. The natural history as well as management depends upon etiology of pulmonary hypertension. Despite newer insights in pathophysiology and management strategies, the outcome of pulmonary veno-occlusive disease remains dismal. The author's report a case of 12-year-old girl who presented with severe pulmonary hypertension. Stepwise evaluation and CT angiographic assessment in the index case led to definitive diagnosis of pulmonary veno-occlusive disease. This case highlights the role of cardiac imaging in localising site of pulmonary hypertension. It is important for health care professionals involved in care of patients with pulmonary hypertension to make correct diagnosis of pulmonary veno occlusive disease for better prognostication as well as to avoid therapeutic mishaps. PMID- 21625839 TI - Clinical pearls in pediatric cardiology. AB - In this section, the authors present four interesting clinical cases that presented with common symptoms, but required careful clinical examination and investigations to arrive at a correct diagnosis and institute appropriate management. The first case is a 3-month-old infant who presented with hypertonia, bulging anterior fontanelle and tachycardia. ECG revealed supraventricular tachycardia which was managed appropriately. The clinical suspicion of meningitis was not borne out by the CSF findings. The second is a pre-school girl who presented with recurrent, episodic wheezing that did not respond to standard asthma therapy. The authors discuss the management approach in such cases; careful examination of radiographs and clinical course led to the correct diagnosis of dilated cardiomyopathy. The next two cases are older children; one of them presented with massive bilateral pleural effusion; this was found to be caused by congestive cardiac failure. The last case is a 10-year-old child presenting with features of right sided heart failure without obvious cardiac abnormality on clinical examination. Advanced investigations confirmed a rare diagnosis of arrhythmogenic right ventricular dysplasia; retrospective ECG examination showed the characteristic findings of this condition. PMID- 21625840 TI - Glucose tolerance in overweight and obese North Indian adolescents. AB - The present analytic study was planned to study the impact of overweight and obesity on glucose tolerance in adolescents. Thirty overweight (BMI analogue >= 25-29.9 kg/m(2)) and 30 obese (BMI analogue >= 30 kg/m(2)) adolescents were included as cases and 30 healthy age and sex matched adolescents comprised the control group. All the study participants were subjected to oral glucose tolerance test (OGTT) as a measure of glucose tolerance.Means of fasting blood glucose levels in normal, overweight and obese groups were 84.5 mg/dl, 86.7 mg/dl and 94.8 mg/dl respectively. Means of two hourly blood glucose levels in normal, overweight and obese groups were 105.8 mg/dl, 117 mg/dl and 127.1 mg/dl respectively. Oral Glucose tolerance test (OGTT) was normal in all the participants from control group but was impaired in four overweight adolescents. In obese group, OGTT was impaired in eight subjects and one obese child had fasting and 2 hourly blood glucose levels in diabetic range. PMID- 21625841 TI - Circumcision in children. PMID- 21625842 TI - Color of meconium and interleukin-6. AB - OBJECTIVE: To test the hypothesis that the color of meconial fluid is associated with inflammatory biomarkers, by determining C-reactive protein (CRP) and Interleukin-6 (IL-6) in serum from the umbilical cord. METHODS: In this prospective study, the authors selected 30 newborns with meconium-stained amniotic fluid (MSAF): 14 with green/brown 656 R color and 16 with brown/cinnamon 654 R color, and 20 newborns which showed clear amniotic fluid without MSAF (non MSAF); all newborns were from mothers without risk factors for neonatal sepsis. RESULTS: IL-6 concentration from umbilical cord blood, [median of 12.9 pg/mL (interquartile range {IQR} 8.7-31.0)] of MSAF-green/brown 656 R increased significantly (p < 0.05) when compared with IL-6 concentration, [median of 9.2 pg/mL (IQR 7.2-12.2)] of newborns with clear amniotic fluid and without meconium. CRP from MSAF-green/brown 656 R was median of 0.5 mg/mL (IQR 0.0-2.7), and median of 1.0 mg/mL (IQR 0.0-5.5) from clear amniotic fluid, without meconium. CONCLUSIONS: Significant association was found between MSAF-green/brown 656 R and increase in IL-6, with normal CRP values. PMID- 21625843 TI - Magnesium sulfate tocolysis and intraventricular hemorrhage in very preterm infants. AB - OBJECTIVE: To estimate the contributory effect of tocolytic magnesium sulfate (MgSO4) exposure to intraventricular hemorrhage (IVH) in preterm infants born at 23-31 wks gestation to mothers without evidence of pregnancy induced hypertension and/or preeclampsia. METHODS: Cases with IVH and controls without IVH were selected from a population-based cohort of preterm infants admitted from January 2004 through May 2008 to the Level III Neonatal Intensive Care Unit (NICU) at Robert Wood Johnson University Hospital. Cases and controls were matched primarily by exact gestational age in completed weeks and secondarily by the birth weight that was same or similar (+/-100 g). The odds of tocolytic MgSO(4) exposure among the cases and controls was tested in a regression model to control the difference in demographic and clinical factors between the IVH cases (IVH+) and controls without IVH (IVH-). RESULTS: Eighty-nine IVH cases and 89 controls were comparable for parity, mode of delivery, antenatal corticosteroid exposure, and surfactant administration. IVH cases were less likely to have preterm premature rupture of membranes and were more likely to be born with low Apgar scores and require ventilation. Among the IVH cases, 30.3% of infants were exposed to tocolytic MgSO4 as compared to 47.2% of controls (Odds Ratio adjusted 0.471, 95% Confidence Interval 0.241, 0.906). CONCLUSIONS: Among the preterm born infants with gestational age 23-31 wks and IVH, tocolytic MgSO4 exposure was less likely to be observed than in neonates with similar clinical characteristics but without IVH, thereby suggesting that antenatal exposure to MgSO(4) may have a protective effect against IVH. PMID- 21625844 TI - Outcome of in-hospital pediatric cardiopulmonary arrest from a single center in Pakistan. AB - OBJECTIVE: To review the incidence and outcome of in-hospital pediatric cardiopulmonary arrest (CPA). METHODS: This retrospective six-year case series was carried out at the PICU and Pediatric Units of Aga Khan University Hospital (AKUH). All children aged 1 month to 14 years who underwent cardiopulmonary resuscitation from January 2001 through December 2006 were included. Data were recorded according to the Utstein style. The outcome variables were sustained return of spontaneous circulation (initial survival) and hospital discharge (final survival). Factors associated with survival were evaluated using logistic regression analysis. RESULTS: The incidence of CPA was 0.4% of all the admissions. Most of the CPR attempts took place in pediatric intensive care unit (53%) and the most frequent etiology was shock (78%). After initial CPR, the sustained return of spontaneous circulation was achieved in 58 patients (55%). Only 12 patients (11%) were discharged alive from the hospital. The most common initial documented rhythm was bradycardia (78%). Multivariate logistic regression analysis revealed that prolonged duration of CPR (>20 min) was the best predictor of initial and final mortality (p < 0.001). CONCLUSIONS: The survival rate of in hospital pediatric cardiopulmonary arrest in the present report is low. PMID- 21625845 TI - Comparison of a self-rated and a clinician-rated measure for identifying depression among adolescents in a primary-care setting. AB - OBJECTIVE: To compare the diagnostic accuracy of a self-rated and a clinician rated measure of depression for primary care use in school setting by pediatricians. METHODS: Two tools for screening depression were administered to early adolescents in three schools. These included the self-rated Beck Depression Inventory (BDI), pediatrician rated Children's Depression Rating Scale-Revised (CDRS-R), and ICD-10 clinical interview by a psychiatrist as reference standard. These tools were compared for their overall performance using Areas Under the Curve (AUC) of Receiver Operating Characteristic (ROC) curves. The optimal screening threshold score for both tools were identified from their sensitivity and specificity plotted for all threshold scores. For the optimal cut-off scores, the diagnostic accuracy parameters like sensitivity, specificity, predictive values, likelihood ratio and diagnostic odds ratio were calculated using contingency table. RESULTS: The area under the curve for BDI was 0.67 and CDRS was 0.50 suggesting that BDI as a screening tool has better diagnostic accuracy. The optimal screening threshold score for BDI was 18 with a sensitivity of 63 and specificity of 70. For the CDRS-R cut-off score of 59, the sensitivity was 36 and specificity was 82 respectively. Using both tools concurrently improved the diagnostic accuracy. CONCLUSIONS: Using the ROC characteristics and various validity indices, the authors showed that BDI has better sensitivity and CDRS-R a better specificity. It might be prudent to use both these instrument simultaneously to improve the identification of depression in primary care settings like school health clinic. PMID- 21625846 TI - Menstrual disorders and menstrual hygiene practices in higher secondary school girls. AB - OBJECTIVE: To study the menstrual problems and menstrual hygiene practices of adolescent girls in Thiruvananthapuram City Corporation. METHODS: Students of class XI and XII in the age group 15-19 years, belonging to ten Higher Secondary Schools within the Thiruvananthapuram City Corporation area were selected for the study by multistage sampling procedure and screened using a pretested self evaluation questionnaire. RESULTS: Menstrual disorders were reported in 21.1%. The most frequently reported problem during menstruation was dysmenorrhoea (72.4%) followed by oligomenorrhoea (11.3%). Only 11.5% of the girls who had menstrual problems sought treatment and majority from a gynecologist. Out of 81.5% girls who reported vaginal discharge, only 5.7% had abnormal discharge. Menstrual hygiene was adequate in the majority of girls. CONCLUSIONS: Menstrual disorders are common in adolescence and can have significant consequences on future reproductive health. PMID- 21625847 TI - Targeting NF-kappaB and HIF-1 pathways for the treatment of cancer: part II. AB - Hypoxia that originates from disturbed growth of solid tumors initiates a cascade of intracellular events engaging hypoxia-inducible factors, HIF-1 and HIF-2. Overexpression of HIF has been confirmed in solid tumors and was unfortunately accompanied with chemo- and radioresistance observed in many patients. Multiple cellular pathways resulting in HIF activation could be successfully inhibited by use of different kinds of drugs (e.g. topotecan, heat shock protein 90 and mTOR inhibitors, YC-1, pleurotin or 2-methoxyestradiol), which are being subjected into intensive investigation in clinical trials. PMID- 21625848 TI - Targeting NF-kappaB and HIF-1 pathways for the treatment of cancer: part I. AB - The process of chronic inflammation is a common link which connects different kinds of environmental pollutants and infections with tumorigenesis. Transcription factor NF-kappaB is a common final target for many inflammatory and cell proliferation pathways, independent of the source of stimuli (e.g., cytokines, growth factors, environmental carcinogens, radiation, hypoxia, bacteria, and viruses). Over-activation of NF-kappaB has been confirmed in many tumors, resulting in worse prognosis for patient survival. Therefore, inhibition of cellular pathways for NF-kappaB activation is nowadays considered as a promising anti-cancer therapy and is extensively studied in clinical trials, or even has been adopted as an approved therapy in some kinds of cancer. PMID- 21625849 TI - Genome-wide analysis of the heat shock transcription factors in Populus trichocarpa and Medicago truncatula. AB - Research has provided substantial evidences that heat shock proteins (HSPs) play essential roles in extreme physiological conditions. Heat shock transcription factors (HSFs) are important HSPs regulators, but their functions are poorly understood, particularly in Populus and Medicago. In this study, a comprehensive bioinformatics analysis of the HSFs was performed in Populus trichocarpa and Medicago truncatula. Twenty-eight Populus HSFs and 16 Medicago HSFs were identified, and comparative analyzes of the two plants were carried out subsequently. HSFs were divided into three different classes and they were diverse and complicated transcription factors. The results of semi-quantitative RT-PCR in Populus suggested six genes (PtHSF-03, PtHSF-13, PtHSF-15, PtHSF-21, PtHSF-22 and PtHSF-23) were markedly increased by heat stress. The results presented here provide an important clue for cloning, expression and functional studies of the HSFs in Populus and Medicago. PMID- 21625850 TI - Probiotic Dahi containing Lactobacillus acidophilus and Bifidobacterium bifidum alleviates age-inflicted oxidative stress and improves expression of biomarkers of ageing in mice. AB - The potential benefiting effects of probiotic Dahi on age-inflicted accumulation of oxidation products, antioxidant enzymes and expression of biomarkers of ageing were evaluated in mice. Probiotic Dahi were prepared by co-culturing in buffalo milk (3% fat) Dahi bacteria (Lactococcus lactis ssp. cremoris NCDC-86 and Lactococcus lactis ssp. lactis biovar diacetylactis NCDC-60) along with selected strain of Lactobacillus acidophilus LaVK2 (La-Dahi) or combined L. acidophilus and Bifidobacterium bifidum BbVK3 (LaBb-Dahi). Four groups of 12 months old mice (6 each) were fed for 4 months supplements (5 g/day) of buffalo milk (3% fat), Dahi, La-Dahi and LaBb-Dahi, respectively, with basal diet. The activities of catalase (CAT) and glutathione peroxidase (GPx) declined and the contents of oxidation products, thiobarbituric acid reactive substances (TBARS) and protein carbonyls, increased in red blood corpuscles (RBCs), liver, kidney and heart tissues and superoxide dismutase (SOD) activity increased in RBCs and hepatic tissues during ageing of mice. Feeding ageing mice with La-Dahi or LaBb-Dahi increased CAT activity in all the four tissues, and GPx activity in RBCs and hepatic tissue, and a significant decline in TBARS in plasma, kidney and hepatic tissues and protein carbonyls in plasma. Feeding mice with probiotic Dahi also reversed age related decline in expression of biomarkers of ageing, peroxisome proliferators activated receptor-alpha, senescence marker protein-30 (SMP-30) and klotho in hepatic and kidney tissues. The present study suggests that probiotic Dahi containing selected strains of bacteria can be used as a potential nutraceutical intervention to combat oxidative stress and molecular alterations associated with ageing. PMID- 21625851 TI - Isolation and cloning of microRNAs from recalcitrant plant tissues with small amounts of total RNA: a step-by step approach. AB - MicroRNAs (miRNAs) are small RNAs (sRNAs) with approximately 21-24 nucleotides in length. They regulate the expression of target genes through the mechanism of RNA silencing. Conventional isolation and cloning of miRNAs methods are usually technical demanding and inefficient. These limitations include the requirement for high amounts of starting total RNA, inefficient ligation of linkers, high amount of PCR artifacts and bias in the formation of short miRNA-concatamers. Here we describe in detail a method that uses 80 MUg of total RNA as the starting material. Enhancement of the ligation of sRNAs and linkers with the use of polyethylene glycol (PEG8000) was described. PCR artifacts from the amplification of reverse-transcribed sRNAs were greatly decreased by using lower concentrations of primers and reducing the number of amplification cycles. Large concatamers with up to 1 kb in size with around 20 sRNAs/concatamer were obtained by using an optimized reaction condition. This protocol provide researchers with a rapid, efficient and cost-effective method for the construction of miRNA profiles from plant tissues containing low amounts of total RNA, such as fruit flesh and senescent leaves. PMID- 21625852 TI - The role of PRKCH gene variants in coronary artery disease in a Chinese population. AB - The aim of the present study was to assess the influences of PRKCH gene variants (1425G/A and _15) on the risk of coronary artery disease (CAD) in a Chinese population. Our study population consisted of 470 CAD patients and 434 control subjects. The alleles frequencies of the two variants were significantly higher among CAD patients than control subjects (P = 0.001 for 1425G/A and P = 0.001 for _15, respectively). In the CAD group, the A allele carriers of 1425G/A and _15 polymorphisms had higher low-density lipoprotein cholesterol (LDL-C) levels than homozygote G allele carriers (P = 0.001 and P = 0.021, respectively). In a multiple logistic regression model adjusted for age, sex, body mass index (BMI), etc., a markedly increased risk of developing CAD was found in subjects carrying GA or AA genotype (P = 0.005 and P = 0.018, respectively). In conclusion, we observed that there was a remarkable association of minor alleles (1425G/A and _15) in the PRKCH gene with an elevated risk of CAD and increased levels of LDL-C in this Chinese population. PMID- 21625853 TI - A trypsin homolog in amphioxus: expression, enzymatic activity and evolution. AB - Trypsin has been documented in a variety of species including both vertebrates and invertebrates, but little is known about it in amphioxus, a model organism for insights into the origin and evolution of vertebrates. Here we identified a trypsin gene in Branchiostoma japonicum. The cDNA was 978 bp long with an ORF encoding a deduced protein of 272 amino acids. The deduced protein had an N terminal signal peptide of 15 amino acids, a 16 activation peptide with the typical cleavage site Arg/Ile, a Tryp_SPc domain with the catalytic triad His(72) Asp(118)-Ser(215) and the S1 substrate binding residue Asp(209), which are all characteristic of trypsinogens. The recombinant trypsin protein was able to hydrolyse the trypsin prototypic substrate BAEE, which was inhibited by the trypsin-specific inhibitor soybean trypsin inhibitor. Both northern blotting and tissue-section in situ hybridization demonstrated that trypsin gene was expressed in a tissue-specific manner, with most abundant levels in the hepatic caecum, mid gut and ovary. And the whole mount in situ hybridization showed that it began to express in the middle third of the full-length primitive gut in 2-day larvae, where the hepatic caecum will form later during development. Phylogenetic analysis indicated that both amphioxus and ascidian trypsins are more closer to each other than to vertebrate trypsins, suggesting a continuous evolutionary divergence of vertebrate trypsins after split from protochordate/vertebrate common ancestor. PMID- 21625854 TI - Implication of genetic variants near TMEM18, BCDIN3D/FAIM2, and MC4R with coronary artery disease and obesity in Chinese: a angiography-based study. AB - Coronary artery disease (CAD) is multifactorial disease which occurs as a result of the interaction of genetic and environmental factors. Obesity is an independent risk factor for cardiovascular disease. Recent genome-wide association studies have identified several genes associated with obesity in Europeans. We wondered whether these genetic variants were associated with CAD. Three single nucleotide polymorphisms (SNPs) rs7561317 near TMEM18, rs7138803 near BCDIN3D/FAIM2 and rs12970134 near MC4R were examined in 930 Han Chinese subjects based on coronary angiography, using polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP) analysis. There were no significant differences in genotypes and allele distributions of three SNPs between CAD and CAD-free groups. The AA genotype of SNP rs12970134 near MC4R was associated to obesity both in CAD group and CAD-free group in Han Chinese population (P < 0.001, OR = 2.96, 95% CI 2.01-3.73; and P = 0.003, OR = 2.59, 95% CI 1.86-3.19, respectively). Our observations suggest that the polymorphism rs12970134 near MC4R may be associated to the risk of obesity in Han Chinese population. PMID- 21625855 TI - Effects of different elicitors on yield of tropane alkaloids in hairy roots of Anisodus acutangulus. AB - The four tropane alkaloids have played a pivotal role in controlling diseases such as the toxic and septic shock, the organophosphorus poison and the acute lung injury. Here, the elicitation effect of different elicitors on the production of tropane alkaloids and the molecular mechanism of enzyme genes in the pathway was firstly demonstrated in hairy roots of Anisodus acutangulus. The results showed ethanol, methyl jasmonate and Ag(+) could improve the accumulation of tropane alkaloids up to 1.51, 1.13 and 1.08 times after 24 h treatment, respectively (P < 0.05), whereas salicylic acid decreased the average content of tropane alkaloids. Furthermore, expression profile analysis results revealed that up-regulation of hyoscyamine-6b-hydroxylase (AaH6H) and little regulation of tropinone reducase II (AaTR2) elicited by ethanol, increased expression of putrescine N-methyltransferase I (AaPMT1) elicited by Ag(+), elevated expression of tropinone reducase I (AaTR1) elicited by methyl jasmonate, respectively, resulted in tropane alkaloids improvement. Our results showed that hairy root culture of A. acutangulus in combination with elicitors was a promising way for production of tropane alkaloids in the future. PMID- 21625856 TI - Revealing epigenetic patterns in gene regulation through integrative analysis of epigenetic interaction network. AB - Chromatin components and DNA methylation play important roles in regulation of gene expression in mammalian genomes. However, the mechanism underlying how they regulate gene transcription, independently or synergistically, remains largely unknown. We constructed an epigenetic interaction network (EIN) of chromatin components, DNA methylation and gene expression by combining partial correlation coefficient with Pearson correlation coefficient. In EIN, we identified nine direct factors for gene expression. They constitute three interaction modules which synergistically affect gene expression. We introduced a new combination strategy to test how these direct factors in each module regulate gene expression synergistically. We found two inter-attracted patterns and one inter-repulsed patterns among the three modules. Furthermore, we identified 22 indirect factors for gene expression which have effect on gene expression via direct factors. DNA methylation, for example, could regulate gene expression through H3K4me3 and Pol II. Our approach has the potential to help in uncovering inherent relationships between epigenetic factors and gene transcription and guiding experiment. PMID- 21625857 TI - Association of 1347 G/A cytochrome P450 4F2 (CYP4F2) gene variant with hypertension and stroke. AB - Genetic variants of cytochrome P450 4F2 (CYP4F2) gene have been suggested to be risk factors for hypertension, cardiovascular diseases and stroke. In the present case-control study we investigated the association of 1347 G/A polymorphism (rs2108622) in the 11th exon region of CYP4F2 gene with hypertension, ischemic stroke and stroke subtypes classified according to TOAST (Trial of Org 10172 in Acute Stroke Treatment) classification. Five hundred and seven stroke patients (hypertensives: normotensives = 279:228) and four hundred and eighty seven, age and sex matched controls (males: females = 356:131) (hypertensives: normotensives = 148:339) were involved in the study. The genotypes were determined by PCR-RFLP technique. Genotypes were confirmed by subjecting the PCR products to sequencing. Significant difference was observed in the genotypic distribution and allelic frequency between the stroke patients and healthy controls. AA genotype and A allele associated significantly with stroke and hypertension [P = 0.009; OR = 1.59 (95% CI = 1.119-2.283) and P = 0.010; OR = 1.26 (95% CI = 1.056-1.502); P = 0.01; OR = 1.58 (95% CI = 1.11-2.272) and P = 0.010; OR = 1.25(95% CI = 1.054 1.504) respectively]. A stepwise logistic regression analysis confirmed these findings. To establish that this polymorphism is associated with stroke independent of hypertension; we compared stroke patients without hypertension with normotensive controls. Significant difference was observed in genotypic distribution and allelic frequency between the two groups (P = 0.001 and 0.002 respectively). Evaluating the association of this polymorphism with stroke subtypes we found significant associations with cardioembolic stroke (P < 0.001). In conclusion our study suggests that 1347A allele of CYP4F2 gene is an important risk factor for hypertension and ischemic stroke. PMID- 21625858 TI - Molecular cloning and its expression of trachealess gene (As-trh) during development in brine shrimp, Artemia sinica. AB - Basic helix-loop-helix-PAS (bHLH-PAS) family transcription factors are implicated in multiple developmental and physiological regulatory processes. Herein, a full length cDNA encoding a bHLH-PAS domain transcription factor trachealess gene (designated as As-trh) was cloned and characterized from brine shrimp (Artemia sinica) for the first time. The full-length cDNA of As-trh was 2,698 bp with a 2,319 bp open reading frame encoding a deduced protein of 772 amino acid polypeptide with a calculated molecular mass of 86.02 kDa and an isoelectric point of 5.87. Sequence alignment revealed that As-trh had high homology with other species trh gene, including the D-trh gene in Drosophila melanogaster and Bm-trh in Bombyx mori. The early and persistent expression of As-trh in the naupliar stages by whole-mount embryonic in situ hybridization and immunohistochemistry suggest that As-trh functions very early in the salt gland and may be required continuously in this tissue. Later in development, expression of As-trh begins to decrease and disappear in salt gland of the older nauplius and appears in the thoracic epipods of the sub-adult Artemia. These results indicated that As-trh might play an important role in osmoregulatiory organ development from the larvae stages through adult stages. PMID- 21625859 TI - Association between type 2 diabetes and CDKN2A/B: a meta-analysis study. AB - Cyclin-dependent kinase inhibitor-2A/B (CDKN2A/B) has been reported as a candidate gene of type 2 diabetes (T2D) based on its chromosomal position and its important role in beta-cell function and regeneration. However, studies to date have reported inconsistent findings regarding the association between T2D and CDKN2A/B. To clarify this inconsistence, we conducted a meta-analysis based on alleles and genotypes prevalence of rs10811661 and rs564398 in CDKN2A/B. The PubMed, EMBASE, and Medline databases were systematically reviewed for studies published between January, 2006, and November, 2010. A total of 35 reports were collected, among of them only 16 studies (including 24,407 cases and 33,937 controls) match the inclusion criteria and were selected for the statistical test. In the meta-analysis of published data, our results suggest that the rs10811661 T allele (OR 1.28, 95% CI 1.21-1.36, P < 1 * 10(-5)) and TT genotype (OR 1.32, 95% CI 1.22-1.43, P < 1 * 10(-5)) of CDKN2A/B were associated with type 2 diabetes respectively, but rs564398 was not (for allele only: OR 0.96, 95% CI 0.88-1.05, P = 0.35). The association between rs10811661 T allele and T2D was observed both in Asia (P < 1 * 10(-4)) and Europe ethnicity groups (P = 0.002). This meta-analysis yielded evidence that rs10811661 of CDKN2A/B confers risk for T2D. Larger studies with mixed ethnicity subjects are required to validate our findings. PMID- 21625860 TI - Selection of reference genes for normalization of qRT-PCR analysis of differentially expressed genes in soybean exposed to cadmium. AB - Accurate normalization of gene expression with qRT-PCR depends on the use of appropriate reference genes (RGs) for the species under a given set of experimental conditions. Multiple RGs for gene expression analysis of soybean exposed to heavy metal stress treatment have not been reported in the literature. In this study, we evaluated the expression stability of ten candidate RGs in leaves, roots and stems of two soybean cultivars exposed to cadmium (Cd). Based on the geNorm and NormFinder analysis, ACT3, PP2A, ELF1B and F-box were the most stable RGs in these gene expression studies. In contrast, G6PD, UBC2, TUB, and ELF1A were the most variable ones and should not be used as RGs in these experimental conditions. PMID- 21625861 TI - Comprehensive gene and microRNA expression profiling reveals the crucial role of hsa-let-7i and its target genes in colorectal cancer metastasis. AB - Accumulating evidence has demonstrated that miRNAs play important roles in the occurrence and development of colorectal cancer (CRC). However, whether miRNAs are associated with the metastasis of CRC remains largely unexplored. The aim of the current study is to profile miRNAs in different CRC metastatic cell lines to identify the biomarkers in CRC metastasis. Gene and miRNA expression profiling was performed to analyze the global expression of mRNAs and miRNAs in the four human CRC cell lines (LoVo, SW480, HT29 and Caco-2) with different potential of metastasis. Expression patterns of mRNAs and miRNAs were altered in different CRC cell lines. By developing an integrated bioinformatics analysis of gene and miRNA expression patterns, hsa-let-7i was identified to show the highest degree in the microRNA-GO-network and microRNA-Gene-network. The expression level of hsa-let-7i was further validated by qRT-PCR in CRC cells. In addition, the targets of hsa let-7i were predicted by two programs TargetScan and PicTar, and target genes were validated by expression profiling in the most epresentative LoVo and Caco-2 cell lines. Eight genes including TRIM41, SOX13, SLC25A4, SEMA4F, RPUSD2, PLEKHG6, CCND2, and BTBD3 were identified as hsa-let-7i targets. Our data showed the power of comprehensive gene and miRNA expression profiling and the application of bioinformatics tools in the identification of novel biomarkers in CRC metastasis. PMID- 21625862 TI - Genetic variation and population structure of swimming crab (Portunus trituberculatus) inferred from mitochondrial control region. AB - Genetic variation and population structure in Portunus trituberculatus along the coast of China were revealed according to 617 bp of mitochondrial DNA control region. 90 polymorphic sites defined 53 distinct haplotypes, showing a moderately high diversity among 72 individuals sampled from eight localities. Neighbor joining tree, statistics analyses of gene flow and genetic differentiation index indicated two populations from Beihai and Laizhou had differentiated. The population from Yingkou, Dandong, Laizhou and Beihai had smaller genetic diversity compared to that from Ningbo, Lianyungang, Qingdao and Japan according to the genetic distance. And mantel test showed significant positive correlation between genetic distance and geographic distance for P. trituberculatus. TCS parsimony network suggested that all the animals sampled were probably the result of recent divergence from a common ancestral haplotype but for Laizhou population. Moreover, the haplotype distribution appeared to correlate with a recent colonization followed by localized genetic differentiation. Mismatch distribution results suggested that Ningbo, Yingkou, Qingdao, Lianyungang and Japan populations, particularly Dandong population had experienced a sudden demographic or spatial expansion. The Pleistocene glaciations might contribute to this process. PMID- 21625863 TI - CDH1 promoter polymorphism and stomach cancer susceptibility. AB - The relationship of stomach cancer susceptibility and the presence of E-cadherin (CDH1) promoter -160 C/A polymorphism had been reported with conflicting results. To further explore the association of this polymorphism with stomach cancer susceptibility, we performed an extensive search of relevant studies and carried out a meta-analysis to obtain a more precise estimate. A total of 16 studies including 2,611 cases and 3,788 controls were involved in this meta-analysis. When all studies involved, the meta-analysis results suggest no statistically significant association between CDH1 -160 C/A polymorphism and stomach cancer risk (CA vs. CC: OR = 1.01, 95% CI: 0.85-1.19; AA vs. CC: OR = 1.05, 95% CI: 0.75 1.46; dominant model: OR = 1.02, 95% CI: 0.86-1.20; recessive model: OR = 1.04, 95% CI: 0.76-1.41). When subgroup analyses were performed by ethnicity, the A allele carriers conferred a decreased stomach cancer risk in Asians (AA vs. CC: OR = 0.67, 95% CI: 0.47-0.96; dominant model: OR = 0.85, 95% CI: 0.72-0.99), but no statistically significant association was found in Caucasians. In conclusion, this meta-analysis suggests that CDH1 -160 A-allele may play a protective role of stomach cancer development in Asians but not in Caucasians. PMID- 21625864 TI - Forensic species identification of large macaws using DNA barcodes and microsatellite profiles. AB - Using mitochondrial and nuclear markers species identification was conducted in the case of seized feathers. Earlier, we had sequenced cytochrome c oxidase subunit I (COI) both from 10 seized specimens and 43 validation specimens from captive macaws belonging to 4 Ara species (A. macao, A. chloropterus, A. ararauna, and A. ambiguus) and identified 19 haplotypes based on COI sequences. Species-level identification using Barcode of Life Data Systems showed that seized feathers shared the highest similarity with scarlet macaws (A. macao), and this result was supported by the tree-base identification with high bootstrap values. Moreover, microsatellite profiles in AgGT17 locus showed that patterns of allelic distribution in the seized feathers were apparently distinct from those of red-and-green macaw (A. chloropterus), but were overlapped with those of A. macao, suggesting that all of seized feathers were derived from several individuals of A. macao. We also determined the parentage of hybrid macaws by the combination of COI barcodes and microsatellite profiles. The technique presented here will contribute to forensic identification and future conservation of large macaws that have been lost due to deforestation. PMID- 21625865 TI - A genetic variant in microRNA-196a2 is associated with increased cancer risk: a meta-analysis. AB - MicroRNAs (miRNAs) are small non-coding RNA molecules that function as negative regulators of gene expression. Common genetic variants (single nucleotide polymorphisms, SNPs) in miRNA genes may alter their expression or maturation resulting in varied functional consequences. Until now, several studies had evaluated the association between the polymorphisms in the hsa-miR-196a2 rs11614913 and cancer risk in diverse populations and in multiple types of cancer, with contradictory outcomes. Therefore, here we performed a meta-analysis to address the association between this polymorphism and cancer risk. A total of nine studies involving 6,540 cases and 7,562 controls were retrieved based on PubMed. Our analysis demonstrated that hsa-miR-196a2 rs11614913 CC genotype significantly increased the cancer risk in homozygote comparison model compared to TT genotype (OR=1.18; 95% CI, 1.01-1.68). Moreover, significant association of this polymorphism with breast cancer was found based on homozygote comparison model (OR=1.30; 95% CI, 1.01-1.26) and dominant model (OR=1.11; 95% CI, 1.01 1.23). In addition, hsa-miR-196a2 rs11614913 CC genotype was significantly associated with cancer risk in Chinese and Indian (OR=1.21; 95% CI, 1.05-1.40), but not in Caucasians (OR=1.03; 95% CI, 0.89-1.19). Taken together, our results indicate that the polymorphism of hsa-miR-196a2 rs11614913 is associated with cancer susceptibility, especially with breast cancer and in Chinese and Indian populations. PMID- 21625867 TI - Gigantic intrapericardial bronchogenic cyst. PMID- 21625866 TI - Crystal structure of a putative transcriptional regulator SCO0520 from Streptomyces coelicolor A3(2) reveals an unusual dimer among TetR family proteins. AB - A structure of the apo-form of the putative transcriptional regulator SCO0520 from Streptomyces coelicolor A3(2) was determined at 1.8 A resolution. SCO0520 belongs to the TetR family of regulators. In the crystal lattice, the asymmetric unit contains two monomers that form an Omega-shaped dimer. The distance between the two DNA-recognition domains is much longer than the corresponding distances in the known structures of other TetR family proteins. In addition, the subunits in the dimer have different conformational states, resulting in different relative positions of the DNA-binding and regulatory domains. Similar conformational modifications are observed in other TetR regulators and result from ligand binding. These studies provide information about the flexibility of SCO0520 molecule and its putative biological function. PMID- 21625868 TI - Optimal gradient reduction after alcohol septal ablation: a case report with anatomical and practical determinants. PMID- 21625869 TI - [Zenker's diverticulum: pro operation]. AB - In case of Zenker's diverticulum, treatment is indicated as soon as the diagnosis is established. Therapy should aim at the elimination of dysphagia and the symptoms of food retention and should reliably prevent recurrence. Currently, three different therapeutic approaches are applied: the classical option is open transcervical myotomy and diverticulectomy/diverticulopexy and alternatively stapled diverticulostomy with a linear stapler or flexible endoscopic diverticulostomy is propagated. As compared to the surgical (open) approach, rigid or flexible endotherapy is less invasive. However, endotherapy is not always feasible for all types of Zenker's diverticulum and the recurrence rate is high. Accordingly, open diverticulectomy is recommended in patients with an adequate life expectancy and good general operability. PMID- 21625870 TI - Heterologous expression characteristics of Trichoderma viride endoglucanase V in the silkworm, Bombyx mori L. AB - Efficient degradation of cellulose needs a synergistic reaction of the cellulolytic enzymes, which include exoglucanases, endoglucanases, and beta-1,4 glucosidase. In this study, we used an improved Bac-to-Bac/BmNPV baculovirus expression system, which lacks the virus-encoded chitinase cathepsin (v-cath) genes of Bombyx mori nucleopolyhedrovirus (BmNPV), to express the endoglucanase V (EG V) gene from Trichoderma viride in silkworm BmN cells and silkworm larvae, and analyzed the characteristics of the recombinant enzyme in silkworm larvae. The result showed that an around 36-kDa protein was visualized in BmN cells at 48 h after the second-generation recombinant mBacmid/BmNPV/EG V baculovirus infection. The crude enzyme extract from the recombinant baculoviruses-infected silkworms exhibited a significant maximum activity at the environmental condition of pH 5.0 and a temperature of 50 degrees C, and increased 39.86% and 37.76% compared with that from blank mBacmid/BmNPV baculovirus-infected silkworms and normal silkworms, respectively. It was stable at pH range from 5.0 to 10.0 and at temperature range from 40 to 60 degrees C. The availability of large quantities of EG V that the silkworm provides might greatly facilitate the future research and the potential application in industries. PMID- 21625871 TI - Biosolubilization of rock phosphate by three stress-tolerant fungal strains. AB - Three stress-tolerant phosphate-solubilizing fungal strains identified as Aspergillus niger, Aspergillus japonicus, and Penicillium simplicissimum were isolated from wheat rhizospheric soil. The strains demonstrated different capabilities of phosphate solubilization in National Botanical Research Institute's phosphate medium containing rock phosphate (RP) as sole phosphorus (P) source, and the solubilization of RP by P. simplicissimum was the most effective among these strains, followed by A. niger and A. japonicus. All the strains exhibited high levels of stress tolerance like 10~45 degrees C temperature, 4~11 pH, 0~3.5% NaCl, and 0~35% PEG 10000. The strains also differed in their abilities to survive and release soluble P from RP under different stresses. A. niger showed significantly higher tolerance to temperature and pH over the other two strains. Higher amount of spores and content of soluble P in the medium were observed in the presence of 3.5% NaCl with P. simplicissimum, followed by A. niger and A. japonicus. P. simplicissimum could not solubilize RP in the presence of 35% PEG 10000, which exhibited the lowest tolerance to desiccation stress among the three strains. PMID- 21625872 TI - Impact of high pyruvate concentration on kinetics of rabbit muscle lactate dehydrogenase. AB - In order to evaluate the effectiveness of L: -lactate dehydrogenase (LDH) from rabbit muscle as a regenerative catalyst of the biologically important cofactor nicotinamide adenine dinucleotide (NAD), the kinetics over broad concentrations were studied to develop a suitable kinetic rate expression. Despite robust literature describing the intricate complexations, the mammalian rabbit muscle LDH lacks a quantitative kinetic rate expression accounting for simultaneous inhibition parameters, specifically at high pyruvate concentrations. Product inhibition by L: -lactate was observed to reduce activity at concentrations greater than 25 mM, while expected substrate inhibition by pyruvate was significant above 4.3 mM concentration. The combined effect of ternary and binary complexes of pyruvate and the coenzymes led to experimental rates as little as a third of expected activity. The convenience of the statistical software package JMP allowed for effective determination of experimental kinetic constants and simplification to a suitable rate expression: [formula: see text] where the last three terms represent the inhibition complex terms for lactate, pyruvate, and pyruvate-NAD, respectively. The corresponding values of K (I-Lac), K (I-Pyr), and K (I-Pyr-NAD) for rabbit muscle LDH are 487.33 mM(-1) and 29.91 mM and 97.47 mM at 22 degrees C and pH 7.8. PMID- 21625873 TI - Production and characterization of a novel extracellular metalloproteinase by a newly isolated moderate halophile, Halobacillus sp. LY6. AB - A moderately halophilic bacterium LY6 with high proteolytic activity was isolated. Biochemical and physiological characterization, along with 16S rDNA sequence analysis placed the isolate in the genus Halobacillus. The salinity of the culture medium strongly influenced the proteinase production of LY6. Maximum enzyme production was observed in the medium containing 5% Na(2)SO(4) or 10% NaCl. Proteinase production was synchronized with bacterial growth and reached a maximum level during the mid-stationary phase. Enzyme purification was carried out by a simple approach including a combination of ammonium sulfate precipitation and Sephacryl S-100 gel filtration chromatography. SDS-PAGE and gelatin zymography analysis revealed it was a monomer with high molecular weight of 69 kDa. Optimal proteinase activity was obtained at pH 10.0, 40 degrees C, and 10% NaCl. It was high active over broad temperature (30-80 degrees C), pH (6.0 12.0), and NaCl concentration (0-25%) ranges, indicating its thermostable, alkali stable, and halotolerant nature. Moreover, the enzyme activity was markedly enhanced by Ca(2+) and Cu(2+), but strongly inhibited by EDTA, PAO, and DEPC, indicating that it probably was a metalloproteinase with cysteine and histidine residues located in its active site. PMID- 21625874 TI - Cloning and characterization of a novel CoA-ligase gene from Penicillium chrysogenum. AB - A novel phenylacetic acid (PAA)-induced CoA-ligase-encoding gene, designated as phlC, has been cloned from penicillin-producing fungus Penicillium chrysogenum. The open reading frame of phlC cDNA was 1671 bp and encoded a 556 amino acid residues protein with the consensus AMP binding site and a peroxisomal targeting signal 1 on its C terminus. The deduced amino acid sequence showed 37% and 38% identity with characterized P. chrysogenum Phl and PhlB protein, respectively. Functional recombinant PhlC protein was overexpressed in Escherichia coli. The purified recombinant enzyme was capable to convert PAA into its corresponding CoA ester with a specific activity of 129.5 +/- 3.026 pmol/min per mg protein. Similar to Phl and PhlB, PhlC displayed broad substrate spectrum and showed higher activities to medium- and long-chain fatty acids. The catalytic properties of PhlC have been determined and compared to those of Phl and PhlB. PMID- 21625875 TI - Screening of Staphylococcus aureus nasal strains isolated from medical students for toxin genes. AB - Three hundred twenty-one students (156 students with no clinical exposure and 165 students with clinical exposure) were screened for nasal colonization by Staphylococcus aureus; 20.9% of students were S. aureus nasal carriers, and 40.3% of S. aureus isolates harbored toxin genes. The most prevalent genes were tst (15.0 %) and sec (13.4 %). Isolates with multiple genes were only found among clinical students (p = 0.045). Six of 11 PFGE clones were positive for toxin genes. Methicillin-resistant (MRSA) isolates were only detected in the clinical students (4.5 %). The exposure of students to the hospital environment neither radically increased S. aureus nasal carriage, nor the frequency of clinically important toxin gene presence, but it could have influenced the positive selection of toxigenic MRSA strains. PMID- 21625876 TI - The influence of fermentation conditions and recycling on the phospholipid and fatty acid composition of the brewer's yeast plasma membranes. AB - Phospholipid (PL) and fatty acid (FA) compositions of the plasma membrane (PM), as well as the FA composition of the PM phosphatidylcholine (PtdCho) and phosphatidylethanolamine (PtdEtn) in the pure culture (zero generation) and the first three recycled generations of the bottom-fermenting brewer's yeast, have been determined. The PL composition differed markedly among the generations; in the zero generation, phosphatidylinositol (PtdIns) was the main PL, accounting for 27% of total PLs, followed by phosphatidic acid and PtdCho. In all recycled generations, the main PL was PtdCho with a marked increase in the first generation compared with the zero (32% and 20%, respectively), followed by PtdIns in the first and second generations. In the FA composition of the PM, 22 FAs were identified, ranging from C(10) to C(26). The compositions of the PM FAs, as well as those of PtdCho and PtdEtn, were characterised by a high preponderance of C(16) acids. Saturated FAs prevailed in the zero generation, whilst unsaturated prevailed in the first and second generation. Although the profiles of FAs in PtdCho and PtdEtn were similar, some marked differences were observed, pointing out to their specific functions in the regulation of membrane properties. PMID- 21625877 TI - The influence of external factors on bacteriophages--review. AB - The ability of bacteriophages to survive under unfavorable conditions is highly diversified. We summarize the influence of different external physical and chemical factors, such as temperature, acidity, and ions, on phage persistence. The relationships between a phage's morphology and its survival abilities suggested by some authors are also discussed. A better understanding of the complex problem of phage sensitivity to external factors may be useful not only for those interested in pharmaceutical and agricultural applications of bacteriophages, but also for others working with phages. PMID- 21625878 TI - Isolation and phylogenetic classification of culturable psychrophilic prokaryotes from the Collins glacier in the Antarctica. AB - Culturable psychrophilic prokaryotes were obtained of samples of glacier sediment, seaside mud, glacier melted ice, and Deschampsia antarctica rhizosphere from Collins glacier, Antarctica. The taxonomic classification was done by a culture-dependent molecular approach involving the Amplified Ribosomal DNA Restriction Analysis. Two hundred sixty colonies were successfully isolated and sub-cultivated under laboratory conditions. The analysis showed a bacterial profile dominated by Beta-proteobacteria (35.2%) followed by Gamma-proteobacteria (18.5%), Alpha-proteobacteria (16.6%), Gram-positive with high GC content (13%), Cytophaga-Flavobacterium-Bacteroides (13%) and Gram-positive with low GC content (3.7%). Eleven of the isolates have been reported previously and the others microorganisms remain uncharacterized. The isolated microorganisms here could be a potential source for biotechnological products, such as cold-active enzymes and secondary metabolites. PMID- 21625880 TI - Renal function and nonvertebral fracture risk in multiethnic women: the Women's Health Initiative (WHI). AB - To examine the association between renal function and fracture in multiethnic women, we studied postmenopausal women enrolled in the Women's Health Initiative. Postmenopausal White women with mild renal dysfunction were at increased risk of nonvertebral fracture; this association was at least partially explained by effects of renal dysfunction on chronic inflammation. Reduced renal function appeared to increase fracture risk among Black women, but there was little evidence to support this association among other racial/ethnic groups. INTRODUCTION: The purpose of this study was to determine whether renal function is associated with fracture risk within racial/ethnic groups. METHODS: A nested case-control study was conducted among 93,673 postmenopausal women; incident nonvertebral fractures were identified in 362 Black, 183 Hispanic, 110 Asian, and 45 American-Indian women. A random sample of 395 White women with incident nonvertebral fracture was chosen. One nonfracture control for each case was selected (matched on age, race/ethnicity, and blood draw date). Cystatin C levels were measured using baseline serum, and estimated glomerular filtration rate calculated (eGFR(cys-c)). RESULTS: Each 1 SD increase in cystatin C was associated with a 1.2-fold increased risk of fracture among White women (adjusted odds ratios [OR], 1.23; 95% confidence intervals [CI], 1.04-1.46). The OR of fracture was 1.16 (95% CI, 0.85-1.58) among women with eGFR(cys-c) 60-90 mL/min/1.73 m(2) and 2.46 (95% CI, 1.16-5.21) among those with eGFR(cys-c) <60 mL/min/1.73 m(2) compared to the reference group (eGFR(cys-c) >90 mL/min/1.73 m(2)) (p trend = 0.05). The association was reduced after adjustment for cytokine TNFalpha soluble receptors (OR, 1.62; 95% CI, 0.59-4.46 for eGFR(cys-c) <60 mL/min/1.73 m(2)). Among Blacks, there was an association between cystatin C and fracture risk (OR per 1 SD increase, 1.15; 95% CI, 1.00-1.32); after adjustment, this association was only modestly attenuated, but no longer statistically significant. There was no evidence of significant associations among Hispanic, Asian, or American-Indian women. CONCLUSION: Postmenopausal White women with mild renal dysfunction are at increased risk of nonvertebral fracture. Effects of renal function on chronic inflammation may mediate this association. Reduced renal function may increase fracture risk among Black women, but there was little evidence to support this association among other racial/ethnic groups. PMID- 21625881 TI - Exercise and fractures in postmenopausal women: 12-year results of the Erlangen Fitness and Osteoporosis Prevention Study (EFOPS). AB - This trial is the first exercise study that focuses on fracture incidence as a primary study endpoint. Although we marginally failed to determine significant effects on "overall" fracture risk (p = .074) or rate ratio (p = .095), our findings further increased the evidence that exercise relevantly prevents fractures in the elderly. INTRODUCTION: The purpose of this study is to determine the effect of strictly supervised long-term exercise training on "overall" fracture incidence and bone mineral density (BMD) in postmenopausal osteopenic women. METHODS: Eighty-five early postmenopausal (1-8 years), osteopenic women living in the area of Erlangen-Nuremberg, Germany without any medication or diseases affecting bone metabolism were assessed after 12 years of supervised exercise (EG) or unvarying lifestyle (control, CG). Exercisers were encouraged to perform two group sessions/week and two home training sessions/week. Calcium and vitamin D supplementation was provided for both groups. "Overall" fractures were determined by questionnaires and structured interviews. The BMD was assessed at lumbar spine and proximal femur by dual-energy X-ray absorptiometry. RESULTS: "Overall" fracture risk ratio in the EG was 0.32 (95% confidence interval (CI), 0.08 to 1.05; p = .074), and the rate ratio for "overall" fractures was 0.38 (95% CI, 0.11 to 1.15; p = .095). BMD changes at lumbar spine (EG, -0.8%; 95% CI, 0.8% to -2.7% vs. CG, -4.0%; 95% CI, -2.4% to -5.7%; p = .011) and femoral neck (EG, 3.7%; 95% CI, -2.4% to -5.0% vs. CG, -6.7%; 95% CI, -5.3% to -8.2%; p = .003) significantly differed between both groups. CONCLUSION: Although we marginally failed to determine significant effects on overall fracture risk or rate ratio, our study increased the body of evidence for the fracture prevention efficiency of exercise programs, with special regard on bone strength (as assessed by bone mineral density measurement). Future studies should focus on subjects more prone to fractures to generate enough statistical power to clearly determine this issue. PMID- 21625882 TI - Comparative trends in hospitalizations for osteoporotic fractures and other frequent diseases between 2000 and 2008. AB - In Switzerland, the number, incidence, and cost of acute hospitalizations for major osteoporotic fractures (MOF) and major cardiovascular events (MCE) increased in both women and men between 2000 and 2008, although the mean length of stay (LOS) was significantly reduced. Similar trend patterns were observed for hip fractures and strokes (decrease) and nonhip fractures and acute myocardial infarctions (increase). INTRODUCTION: The purpose of this study was to compare the trends and epidemiological characteristics of hospitalizations for MOF and other frequent diseases between years 2000 and 2008 in Switzerland. METHODS: Trends in the number, age-standardized incidence, mean LOS, and cost of hospitalized MOF and MCE (acute myocardial infarction, stroke, and heart failure) were compared in women and men aged >= 45 years, based on data from the Swiss Federal Statistical Office. RESULTS: Between 2000 and 2008, the incidence of acute hospitalizations for MOF increased by 3.4% in women and 0.3% in men. In both sexes, a significant decrease in hip fractures (-15.0% and -11.0%) was compensated by a concomitant, significant increase in nonhip fractures (+24.8% and +13.8%). Similarly, the incidence of acute hospitalizations for MCE increased by 4.4% in women and 8.2% in men, as an aggregated result from significantly increasing acute myocardial infarctions and significantly decreasing strokes. While the mean LOS in the acute inpatient setting decreased almost linearly between years 2000 and 2008 in all indications, the inpatient costs increased significantly (p < 0.001) for MOF (+30.1% and +42.7%) and MCE (+22.6% and +47.1%) in women and men, respectively. CONCLUSIONS: Between years 2000 and 2008, the burden of hospitalized osteoporotic fractures to the Swiss healthcare system has continued to increase in both sexes. In women, this burden was significantly higher than that of MCE and the gap widened over time. PMID- 21625883 TI - The rate of hip osteoarthritis in patients with proximal femoral fractures versus hip contusion. AB - We found no difference in the rate of radiological hip osteoarthritis in the injured hip when comparing 349 patients with proximal femoral fractures and 112 patients with hip contusion. There was, however, a tendency for more osteoarthritis in patients with trochanteric fractures than in patients with femoral neck fractures. INTRODUCTION: Osteoarthritis (OA) and osteoporotic fractures are two age-related disorders associated with considerable morbidity. There is a clinical impression of an inverse relation between osteoarthritis and osteoporosis, and a protective effect of OA against osteoporotic fractures has been proposed. METHODS: We performed a case-control study in 461 subjects. Cases (n = 349) were patients aged 50 years or above who sustained a proximal femoral fracture from November 2003 to October 2004, registered prospectively in the department's fracture register. Controls (n = 112) were patients aged 50 years or above with the diagnosis of hip contusion, recruited from the hospital's discharge register. Radiographic OA was scored according to Kellgren and Lawrence (K&L), and minimal joint space (MJS) was measured in both hips when possible. A K&L grade II or higher or an MJS less than 2.5 mm was defined as OA. RESULTS: Both in the hip fracture group and in the contusion group mean, the MJS was 3.5 mm on the injured side (p = 0.79). In the fracture group, 31/250 (12%) had MJS <2.5 mm and 16/112 (14%) in the contusion group (p = 0.18). In the fracture group, 40/250 (16%) had a K&L OA grade II or higher, and in the contusion group 20/112 (18%) persons had a K&L OA grade II or higher (p = 0.66). There was a tendency for a higher incidence of OA in patients with trochanteric fractures compared with patients with cervical fractures. CONCLUSIONS: We found no differences on the injured side in the rate of hip OA between hip fracture patients and hip contusion patients. PMID- 21625884 TI - Genetic variants in the SOX6 gene are associated with bone mineral density in both Caucasian and Chinese populations. AB - SUMMARY: Given the biological function of SOX6 and recent genome-wide association finding, we performed a fine-mapping association analyses to investigate the relationship between SOX6 and BMD both in Caucasian and Chinese populations. We identified many single-nucleotide polymorphisms (SNPs) within or near the SOX6 gene to be significantly associated with hip bone mineral density (BMD). INTRODUCTION: SOX6 gene is an essential transcription factor in chondrogenesis and cartilage formation. Recent genome-wide association studies (GWAS) detected a SNP (rs7117858) located at the downstream of SOX6 significantly associated with hip BMD. METHODS: Given the biological function of SOX6 and the GWAS finding, we considered SOX6 as a new candidate for BMD and osteoporosis. Therefore, in this study, we performed a fine-mapping association analyses to investigate the relationship between SNPs within and near the SOX6 gene and BMD at both hip and spine. A total of 301 SNPs were tested in two independent US Caucasian populations (2,286 and 1,000 unrelated subjects, respectively) and a Chinese population (1,627 unrelated Han subjects). RESULTS: We confirmed that the previously reported rs7117858-A was associated with reduced hip BMD, with combined P value of 2.45 * 10(-4). Besides this SNP, we identified another 19 SNPs within or near the SOX6 gene to be significantly associated with hip BMD after false discovery rate adjustment. The most significant SNP was rs1347677 located at the intron 3 (P = 3.15 * 10(-7)). Seven additional SNPs in high linkage disequilibrium with rs1347677 were also significantly associated with hip BMD. SNPs in SOX6 showed significant skeletal site specificity since no SNP was detected to be associated with spine BMD. CONCLUSION: Our study identified many SNPs in the SOX6 gene associated with hip BMD even across different ethnicities, which further highlighted the importance of the SOX6 gene influencing BMD variation and provided more information to the understanding of the genetic architecture of osteoporosis. PMID- 21625885 TI - Changes in femur neck bone density in US adults between 1988-1994 and 2005-2008: demographic patterns and possible determinants. AB - SUMMARY: This analysis compares femur neck bone mineral density (FNBMD) and bone determinants in adults between National Health and Nutrition Examination Survey (NHANES) III (1988-1994) and NHANES 2005-2008. FNBMD was higher in NHANES 2005 2008 than in NHANES III, but between-survey differences varied by age, sex, and race/ethnicity. The likelihood that FNBMD has improved appears strongest for older white women. INTRODUCTION: Recent data on hip fracture incidence and femur neck osteoporosis suggest that the skeletal status of older US adults has improved since the 1990s, but the explanation for these changes remains uncertain. METHODS: The present study compares mean FNBMD of adults ages 20 years and older between the third (NHANES III, 1988-1994) and NHANES 2005-2008. Dual energy X-ray absorptiometry systems (pencil beam in NHANES III, fan beam in NHANES 2005-2008) were used to measure hip BMD, and several bone determinants are compared between surveys to assess their potential role in explaining observed FNBMD differences. RESULTS: FNBMD was higher overall in NHANES 2005-2008 than in NHANES III, but between-survey differences varied by age, sex, and race/ethnicity. Although FNBMD differences in several groups were small enough (<=3%) to be attributable to use of different dual-energy X-ray absorptiometry (DXA) systems in the two surveys, variability in size and direction of the differences does not support artifactual differences in DXA methodology as the sole explanation. Several FNBMD determinants (body size, smoking, selected bone active medications, self-reported health status, calcium intake, and caffeine consumption) changed in a bone-improving direction in older adults, but FNBMD in older non-Hispanic white women remained significantly higher in 2005-2008 even after adjusting for DXA methodology or for the selected bone determinants. CONCLUSION: The likelihood that FNBMD has improved appears strongest for older white women, but the reason for the improvement in this group remains unclear. PMID- 21625886 TI - Treatment satisfaction and persistence among postmenopausal women on osteoporosis medications: 12-month results from POSSIBLE USTM. AB - SUMMARY: Women in POSSIBLE USTM who expressed greater treatment satisfaction at study entry were more likely to persist with osteoporosis therapy over a 1-year period. Lower satisfaction among women with moderate/severe side effects increased the risk of discontinuation/switching by 67%. Treatment satisfaction and side effect experience influence osteoporosis medication adherence. INTRODUCTION: Non-adherence is common among women using postmenopausal osteoporosis (PMO) medications. We describe the association between treatment satisfaction, measured with the Treatment Satisfaction Questionnaire for Medication (TSQM), and the risk of discontinuation/switching PMO medications using patient-reported data from a large, longitudinal cohort study. METHODS: Data from 2,405 participants in the Prospective Observational Scientific Study Investigating Bone Loss Experience (POSSIBLE USTM) Study were evaluated. Cox proportional hazards regression was used to estimate hazard ratios (HR) for the association between treatment satisfaction at study entry and self-reported discontinuation/switching of pharmacologic PMO medications over a 1-year follow up period. Logistic regression was used to evaluate relationships between treatment satisfaction, lifestyle behaviors, and compliance with bisphosphonate dosing instructions. RESULTS: Median TSQM scores were highest (indicating greatest satisfaction) for the side effects domain [n = 1,182; median = 87.5 (Q1 = 75.0, Q3 = 100.0)] and lowest for global satisfaction [n = 2,340; median = 64.0 (Q1 = 55.7, Q3 = 77.7)]. Median scores decreased for the side effects and global satisfaction domains as patient-reported side effect severity increased. Women with higher satisfaction were less likely to discontinue/switch medications than women with lower scores (adjusted HRs for convenience 0.73, 95% CI = 0.63-0.85; effectiveness 0.82, 95% CI = 0.70-0.97; and global satisfaction 0.73, 95% CI = 0.63-0.85). Lower treatment satisfaction was particularly influential among women who reported moderate/severe side effects (HR = 0.60, 95% CI = 0.37-0.97). CONCLUSIONS: Lower treatment satisfaction was associated with a 22% (1/0.82) to 67% (1/0.60) increased risk of discontinuation/switching osteoporosis medication during 1 year of follow-up. PMID- 21625887 TI - Suppression of bone turnover by B-cell depletion in patients with rheumatoid arthritis. AB - The role of B cells in inflammatory bone formation and resorption is controversial. We investigated this in patients with rheumatoid arthritis (RA) treated with rituximab, a B-cell depleting antibody. We found a significant suppression in bone turnover, possibly a direct effect or as a consequence of a reduction in inflammation and disease activity. INTRODUCTION: RA is the most prevalent inflammatory joint disease, in which B cells play an important role. However, the role of B cells in bone turnover is controversial and RA subjects treated with rituximab, a B-cell depleting monoclonal antibody, provide an ideal model for determining the role of B cells in inflammatory bone resorption. METHODS: Serum from 46 RA patients, collected pre- and post-rituximab therapy, was analysed for biomarkers of bone turnover (procollagen type I amino-terminal propeptide [P1NP], osteocalcin, beta-isomerised carboxy-terminal telopeptide of type 1 collagen [betaCTX] and osteoprotegerin [OPG]). RESULTS: A significant decrease in bone resorption was observed 6 months after rituximab (median change betaCTX -50 ng/L, 95%CI -136, -8 p < 0.001, this equates to -37%; 95%CI -6, -49), mirrored by a reduction in disease activity. Similarly, there was a significant increase in P1NP, a marker of bone formation (median change P1NP 5.0 MUg/L, 95%CI -1.0, 11.2, p = 0.02; 13%; 95%CI -3, 39), but no significant change in osteocalcin or OPG levels. The percentage change from baseline of betaCTX in a subgroup of patients (not on prednisolone or bisphosphonate) was significantly correlated with the percentage reduction in DAS28 score (r (s) = 0.570, p = 0.014). CONCLUSIONS: In conclusion, we have found that B-cell depletion increases bone formation and decreases bone resorption in RA patients; this may be a direct effect on osteoblasts and osteoclasts, respectively, and be at least partially explained by the decreased inflammation and disease activity. PMID- 21625888 TI - High prevalence of vitamin D deficiency among healthy Saudi Arabian men: relationship to bone mineral density, parathyroid hormone, bone turnover markers, and lifestyle factors. AB - SUMMARY: In this cross-sectional study, the prevalence of vitamin D deficiency [serum 25-hydroxyvitamin D (25(OH)D) <50 nmol/L] was 87.8% among Saudi Arabian men. There was a linear inverse relationship between serum 25(OH)D and intact parathyroid hormone (PTH) levels, but without a threshold of 25(OH)D at which intact PTH values plateaued. INTRODUCTION: Vitamin D insufficiency and/or deficiency has now reached epidemic proportions and has been linked to low bone mineral density (BMD), some lifestyle factors, and obesity in adults. This relationship is not well documented in Saudi Arabian men. This study examines the relationship between vitamin D status, intact parathyroid hormone (intact PTH), and lifestyle factors among Saudi Arabian men. METHODS: This cross-sectional study involved 834 men aged 20-74 years living in Jeddah area who were randomly selected and medically examined. Men had their BMD (lumbar spine (L1-L4) and neck femur), 25(OH)D, intact PTH, and other parameters measured according to detailed inclusion criteria. RESULTS: Deficiency (25(OH)D<50 nmol/L) and insufficiency (>=50-75 nmol/L) were present in 87.8% and 9.7%, respectively. Deficiency was common among older and obese men with no education and sedentary lifestyle sampled during summer and spring. Serum 25(OH)D showed an inverse linear relationship with intact PTH, but there was no threshold of serum 25(OH)D at which PTH levels plateaued. There was a positive correlation between BMD values at both lumbar spine (L1-L4) (P < 0.023) and neck femur (P < 0.036) and serum 25(OH)D levels, respectively. CONCLUSIONS: Functionally significant vitamin D deficiency affects BMD and bone turnover markers among Saudi Arabian men and is largely attributed to older age, obesity, sedentary lifestyle, no education, poor exposure to sunlight, smoking, and poor dietary vitamin D supplementation. The data suggest that an increase in PTH cannot be used as a marker for vitamin D deficiency. PMID- 21625889 TI - Validation of the adherence evaluation of osteoporosis treatment (ADEOS) questionnaire for osteoporotic post-menopausal women. AB - SUMMARY: We developed and validated a specific 12-item questionnaire to evaluate adherence to oral antiresorptive medication by post-menopausal osteoporotic women in everyday practice. Over the following 9 months, an index of <=16 was associated with an increase in the risk of treatment discontinuation of 1.69 and of 2.10 for new patients who had started treatment within the previous year. INTRODUCTION: Adherence to medication in osteoporosis is poor. The goal of this study was to develop and validate a disease-specific questionnaire to evaluate adherence to treatment of women with post-menopausal osteoporosis taking oral antiresorptive medication. METHODS: A prototype adherence questionnaire with 45 items developed from patient interview, literature review, and physician opinion was evaluated in a sample of 350 post-menopausal women with osteoporosis treated in primary care. Item responses were matched against scores on the Morisky Medication Adherence Scale (MMAS). The most discriminant items were retained in the final questionnaire. Concurrent and predictive validity were assessed. RESULTS: Twelve items were associated with MMAS score at a probability level of 0.05. These were retained in the final questionnaire which provided an adherence index ranging from 0 to 22. An index of >=20 was associated with a high probability of persistence and an index <= 16 with a high probability of treatment discontinuation in the following 9 months. CONCLUSIONS: The ADEOS-12 is a simple patient-reported measure to determine adherence to osteoporosis treatments with good concurrent and discriminant validity. This is the first disease-specific adherence measure to have been developed for osteoporosis. PMID- 21625890 TI - Physiological relevance and contribution to metal balance of specific and non specific Metallothionein isoforms in the garden snail, Cantareus aspersus. AB - Variable environmental availability of metal ions represents a constant challenge for most organisms, so that during evolution, they have optimised physiological and molecular mechanisms to cope with this particular requirement. Metallothioneins (MTs) are proteins that play a major role in metal homeostasis and as a reservoir. The MT gene/protein systems of terrestrial helicid snails are an invaluable model for the study of metal-binding features and MT isoform specific functionality of these proteins. In the present study, we characterised three paralogous MT isogenes and their expressed products in the escargot (Cantareus aspersus). The metal-dependent transcriptional activation of the three isogenes was assessed using quantitative Real Time PCR. The metal-binding capacities of the three isoforms were studied by characterising the purified native complexes. All the data were analysed in relation to the trace element status of the animals after metal feeding. Two of the three C. aspersus MT (CaMT) isoforms appeared to be metal-specific, (CaCdMT and CaCuMT, for cadmium and copper respectively). A third isoform (CaCd/CuMT) was non-specific, since it was natively recovered as a mixed Cd/Cu complex. A specific role in Cd detoxification for CaCdMT was revealed, with a 80-90% contribution to the Cd balance in snails exposed to this metal. Conclusive data were also obtained for the CaCuMT isoform, which is involved in Cu homeostasis, sharing about 30-50% of the Cu balance of C. aspersus. No apparent metal-related physiological function was found for the third isoform (CaCd/CuMT), so its contribution to the metal balance of the escargot may be, if at all, of only marginal significance, but may enclose a major interest in evolutionary studies. PMID- 21625891 TI - Fanconi-Bickel syndrome in a 3-year-old Indian boy with a novel mutation in the GLUT2 gene. AB - Fanconi-Bickel syndrome is a rare autosomal recessive disorder characterized by hepatorenal glycogen accumulation, proximal renal tubular dysfunction and impaired utilization of glucose and galactose. Most cases have been reported from Europe, Japan, Turkey and the Mediterranean belt. We report a 3-year-old boy from southern India who presented with doll-like facies, florid rickets, massive hepatomegaly, growth retardation, renomegaly and laboratory evidence of proximal renal tubular dysfunction. Liver biopsy demonstrated evidence of glycogenosis. Direct sequencing of genomic DNA confirmed a diagnosis of Fanconi-Bickel syndrome, revealing a G-to-A substitution at position -1 of the splicing acceptor site in intron 1 of the GLUT2 gene in a homozygous pattern (c.16-1G>A or IVS1 1G>A). This novel mutation has not been described in earlier studies. The child was treated with oral potassium citrate, oral phosphorus supplementation, and alpha-calcitriol, on which metabolic derangements were corrected. PMID- 21625892 TI - Long-term beneficial effects of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker therapy for patients with advanced immunoglobulin A nephropathy and impaired renal function. AB - BACKGROUND: There are few reports analyzing the effects of angiotensin-converting enzyme inhibitors (ACEIs) and/or angiotensin receptor blockers (ARBs) on the long term renal survival of advanced immunoglobulin A nephropathy (IgAN) patients. PATIENTS AND METHODS: In this retrospective cohort analysis, we divided 66 IgAN patients with an estimated glomerular filtration rate (eGFR) <60 ml/min into three groups: ACEI group (n = 20, treated with ACEIs), ARB group (n = 23, treated with ARBs), and control group (n = 23, treated with antiplatelet agents), and analyzed the clinical and histological background, renal survival rate until the primary endpoint of 50% decrease of eGFR from baseline, and the secondary endpoint of progression to end-stage renal disease, and the risk factors for progression. RESULTS: The clinical and histological background without serum IgA and C3 were not significantly different among the three groups. The renal survival rate until the primary and secondary endpoints was significantly higher in the ACEI and ARB groups than in the control group. The independent risk factors for progression were higher mean blood pressure (hazard ratio [HR] 1.76, P = 0.04), higher histological grade (HR 2.54, P = 0.0184) at baseline, and without ACEIs or ARBs (HR 7.09, P = 0.001), but decreased proteinuria and blood pressure. The risk factors with resistance to ACEIs or ARBs were higher blood pressure and lower eGFR at baseline. There was no difference regarding the survival rate and the risk for progression between ACEI s and ARBs. CONCLUSION: ACEIs or ARBs were effective for long-term renal survival of advanced IgAN, although proteinuria and blood pressure did not decrease. PMID- 21625893 TI - Safety of erlotinib treatment in outpatients with previously treated non-small cell lung cancer in Japan. AB - PURPOSE: Erlotinib is the first epidermal growth factor receptor-tyrosine kinase inhibitor shown to provide a survival benefit for advanced non-small-cell lung cancer (NSCLC) patients. Adverse drug reactions of erlotinib in Japanese, which may be very different from those in Caucasians because of differences in genetic background, have not been fully reported. Therefore, we aimed to clarify the safety profile of erlotinib. METHODS: Forty-eight patients with pretreated NSCLC were treated with erlotinib between March 2008 and January 2009 in this historical cohort study at Kyoto University Hospital Outpatients Oncology Unit. Erlotinib 150 mg/day was administered until progressive disease or discontinuation due to adverse events. The primary endpoint was frequency and degree of adverse events, and secondary endpoints were clinical efficacy including response rate, disease control rate, progression-free survival and overall survival. RESULTS: Of 48 patients, 3 patients experienced erlotinib induced interstitial pneumonitis, which appeared on day 15 and 70 in 2 patients who recovered and on day 8 in 1 patient who died. The incidences of pruritus, dry skin, diarrhea and stomatitis rapidly increased within 14 days after the start of medication with erlotinib. However, these adverse events were well controllable in outpatients treated with erlotinib. Overall response rate was 10% and disease control rate was 68%. The median progression-free survival was 58 days (95% confidence interval 30-118) and the median overall survival was 229 days (95% confidence interval 135-not available). CONCLUSIONS: Outpatients with NSCLC can be treated with initial administration of erlotinib by careful management. PMID- 21625894 TI - Role of lymph node dissection in managing urologic cancers. PMID- 21625895 TI - NMR and NQR parameters of the SiC-doped on the (4,4) armchair single-walled BPNT: a computational study. AB - The structural properties, NMR and NQR parameters in the pristine and silicon carbide (SiC) doped boron phosphide nanotubes (BPNTs) were calculated using DFT methods (BLYP, B3LYP/6-31G) in order to evaluate the influence of SiC-doped on the (4,4) armchair BPNTs. Nuclear magnetic resonance (NMR) parameters including isotropic (CS(I)) and anisotropic (CS(A)) chemical shielding parameters for the sites of various (13)C, (29)Si, (11)B, and (31)P atoms and quadrupole coupling constant (C ( Q )), and asymmetry parameter (eta ( Q )) at the sites of various (11)B nuclei were calculated in pristine and SiC- doped (4,4) armchair boron phosphide nanotubes models. The calculations indicated that doping of (11)B and (31)P atoms by C and Si atoms had a more significant influence on the calculated NMR and NQR parameters than did doping of the B and P atoms by Si and C atoms. In comparison with the pristine model, the SiC- doping in Si(P)C(B) model of the (4,4) armchair BPNTs reduces the energy gaps of the nanotubes and increases their electrical conductance. The NMR results showed that the B and P atoms which are directly bonded to the C atoms in the SiC-doped BPNTs have significant changes in the NMR parameters with respect to the B and P atoms which are directly bonded to the Si atoms in the SiC-doped BPNTs. The NQR results showed that in BPNTs, the B atoms at the edges of nanotubes play dominant roles in determining the electronic behaviors of BPNTs. Also, the NMR and NQR results detect that the Fig. 1b (Si(P)C(B)) model is a more reactive material than the pristine and the Fig. 1a (Si(B)C(p)) models of the (4,4) armchair BPNTs. PMID- 21625896 TI - The role of CS2 in CS2/NMP mixed solvent in weakening the hydrogen bond of OH-N in coal: a DFT investigation. AB - The interaction processes of trace amounts of N-methyl-2-pyrrolidinone (NMP), CS(2)/NMP (1:1 by volume) and pure NMP solvent with the hydrogen bond of OH?N in coal were constructed and simulated by density functional theory methods. The distances and bond orders between the main related atoms, and the hydrogen bond energy of OH?N were calculated. The calculated results show that pure NMP solvent does not weaken the hydrogen bond of OH?N in coal. However, trace amounts of NMP and CS(2)/NMP (1:1 by volume) have a strong capacity to weaken the hydrogen bond of OH?N in coal. The H2-N3 distances are elongated from 1.87 A to 3.80 A and 3.44 A, the bond orders of H2-N3 all disappear, and the corresponding hydrogen bond energies of OH?N in coal decrease from 45.72 kJ mol(-1) to 7.06 and 11.24 kJ mol( 1), respectively. These results show that CS(2) added to pure NMP solvent plays an important role in releasing the original capacity of NMP to weaken the hydrogen bond of OH?N in coal, in agreement with experimental observations. PMID- 21625897 TI - Topological properties of some PhSeX compounds. AB - A theoretical study on the series of compounds "PhSeX", where Ph = phenyl, Se = selenium and X = Cl, Br, I, CN or SCN, is reported and compared with previously reported experimental data. The molecular geometry for these PhSeX compounds was studied at the DFT/B3LYP level of calculation by means of the 6-311G(d,p) basis set. The equilibrium structures of the molecules were dependent on the method employed to compare the known solid structures. A topological study of the calculated PhSeX species, based on the AIM theory, was carried out to gain a deeper insight into the bonding nature and to find an explanation for the structural diversity exhibited by these PhSeX compounds. The results reported herein illustrate the subtle differences in the solid-state structures of PhSeX compounds. PMID- 21625898 TI - Theoretical study on the structural, vibrational, and thermodynamic properties of the (Br2GaN3)n (n = 1-4) clusters. AB - The molecular geometries, vibrational properties, and thermodynamic properties of the clusters (Br(2)GaN(3))( n )(n = 1-4) were studied at the B3LYP/6-311+G* level. The optimized clusters (Br(2)GaN(3))( n )(n = 2-4) were all found to possess a cyclic structure consisting of Ga atoms bridged by the alpha-nitrogen of the azide groups. A discussion of the relationships between the geometrical parameters and the degree of oligomerization n is provided. Features in the IR spectra were assigned by vibrational analysis. Trends in thermodynamic properties with temperature and degree of oligomerization n are discussed. Thermodynamic analysis of the gas-phase reaction showed that the formation of the clusters (Br(2)GaN(3))( n )(n = 2-4) is thermodynamically favorable considering the enthalpies at 298.2 K. The calculated results for the Gibbs free energies were negative, which indicates that the oligomerizations can occur spontaneously at 298.2 K. PMID- 21625899 TI - Ab initio simulation of the effect of the potential of water on the electronic structure of arginine. AB - As a part of the task of constructing the equivalent potential of water in order to obtain a reliable electronic structure for a protein, the equivalent potential of water for an arginine molecule was constructed by performing first-principles, all-electron, ab initio calculations. The process consisted of three steps. First, the electronic structure of arginine was calculated using a free cluster calculation. Then, the minimum-energy geometric structure of the system Arg(+)+9H(2)O was found using free cluster calculations. Then, based on the optimized geometric structure of the Arg(+)+9H(2)O system, the electronic structure of Arg(+) in the potential of water was calculated using the SCCE method. Finally, by performing SCCE calculations, the effect of water on the electronic structure of Arg(+) was simulated with dipoles. The results show that the effect of water on the electronic structure of Arg(+) is to broaden the energy gap tenfold, and to increase the eight eigenvalues below the HOMO by about 0.0546 Ry on average. The water potential can be accurately simulated using dipoles. PMID- 21625900 TI - Architecture, electronic structure and stability of TM@Ge(n) (TM = Ti, Zr and Hf; n = 1-20) clusters: a density functional modeling. AB - The present study reports the geometry, electronic structure and properties of neutral and anionic transition metal (TM = Ti, Zr and Hf)) doped germanium clusters containing 1 to 20 germanium atoms within the framework of linear combination of atomic orbitals density functional theory under spin polarized generalized gradient approximation. Different parameters, like, binding energy (BE), embedding energy (EE), energy gap between the highest occupied and lowest unoccupied molecular orbitals (HOMO-LUMO), ionization energy (IP), electron affinity (EA), chemical potential etc. of the energetically stable clusters (ground state cluster) in each size are calculated. From the variation of these parameters with the size of the clusters the most stable cluster within the range of calculation is identified. It is found that the clusters having 20 valence electrons turn out to be relatively more stable in both the neutral and the anionic series. The sharp drop in IP as the valence electron count increases from 20 to 21 in neutral cluster is in agreement with predictions of shell models. To study the vibrational nature of the clusters, IR and Raman spectrum of some selected TM@Ge(n) (n = 15,16,17) clusters are also calculated and compared. In the end, relevance of calculated results to the design of Ge-based super-atoms is discussed. PMID- 21625901 TI - Density functional theory studies on the inclusion complexes of cyclic decapeptide with 1-phenyl-1-propanol enantiomers. AB - Cyclic peptides are exciting novel hosts for chiral and molecular recognition. In this work, the inclusion complexes of cyclic decapeptide (CDP) with the 1-phenyl 1-propanol enantiomers (E-PP) are firstly studied using the density functional theory (DFT) B3LYP method. Our calculated results indicated that S(-)-1-phenyl-1 propanol (S-PP) could form a more stable inclusion complex with CDP than that of R(+)-1-phenyl-1-propanol (R-PP). The obvious differences in binding energy and thermodynamics data suggest that the cyclic decapeptide could differentiate the two enantiomers. Furthermore, molecular dynamics simulation results have supported the conclusions obtained by DFT. The current investigation shows that cyclic peptide is a desirable host molecule for chiral and molecular recognition. PMID- 21625902 TI - Problems with molecular mechanics implementations on the example of 4-benzoyl-1 (4-methyl-imidazol-5-yl)-carbonylthiosemicarbazide. AB - Results from force fields implemented in HyperChem, a program frequently used in studies of bioactive compounds, have been compared using the example of the conformational analysis of a 1-carbonylthiosemicarbazide that exhibits strong antibacterial activity. By comparing these results with the original force fields and the experimental NMR ROESY spectrum, it was shown that these implementations can lead to erroneous results. PMID- 21625903 TI - Influence of stereochemistry on proton transfer in protonated tripeptide models. AB - Vectorial proton transfer among carbonyl oxygen atoms was studied in two models of tripeptide via quantum chemical calculations using the hybrid B3LYP functional and the 6-31++G basis set. Two principal proton transfer pathways were found: a first path involving isomerization of the proton around the double bond of the carbonyl group, and a second based on the large conformational flexibility of the tripeptide model where all carbonyl oxygen atoms cooperate. The latter pathway has a rate-determining step energy barrier that is only around half of that for the first pathway. As conformational flexibility plays a crucial role in second pathway, the effect of attaching methyl groups to the alpha carbon atoms was studied. The results obtained are presented for all four possible stereochemical configurations. PMID- 21625904 TI - Unique example of amyloid aggregates stabilized by main chain H-bond instead of the steric zipper: molecular dynamics study of the amyloidogenic segment of amylin wild-type and mutants. AB - Most proteins do not aggregate while in their native functional states. However, they may be disturbed from their native conformation by certain change in the environment, and form unwanted oligomeric or polymeric aggregates. Recent experimental data demonstrate that soluble oligomers of amyloidogenic proteins are responsible for amyloidosis and its cytotoxicity. Human islet amyloid polypeptide (IAPP or amylin) is a 37-residue hormone found as fibrillar deposits in pancreatic extracts of nearly all type II diabetics. In this study we performed in silico mutation analysis to examine the stability of the double layer five strand aggregates formed by heptapeptide NNFGAIL segment from amyline peptide. This segment is one of the shortest fragments that can form amyloid fibrils similar to those formed by the full length peptide. The mutants obtained by single glycine replacement were also studied to investigate the specificity of the dry self-complementary interface between the neighboring beta-sheet layers. The molecular dynamics simulations of the aggregates run for 20 ns at 330 K, the degree of the aggregate disassembly was investigated using several geometry analysis tools: the root mean square deviations of the C(alpha) atoms, root mean square fluctuations per residue, twist angles, interstrand distances, fraction of the secondary structure elements, and number of H-bonds. The analysis shows that most mutations make the aggregates unstable, and their stabilities were dependent to a large extent on the position of replaced residues. Our mutational simulations are in agreement with the pervious experimental observations. We also used free binding energy calculations to determine the role of different components: nonpolar effects, electrostatics and entropy in binding. Nonpolar effects remained consistently more favorable in wild type and mutants reinforcing the importance of hydrophobic effects in protein-protein binding. While entropy systematically opposed binding in all cases, there was no clear trend in the entropy difference between wildtype and glycine mutants. Free energy decomposition shows residues situated at the interface were found to make favorable contributions to the peptide-peptide association. The study of the wild type and mutants in an explicit solvent could provide valuable insight into the future computer guided design efforts for the amyloid aggregation inhibitor. PMID- 21625905 TI - Molecular electrostatic potentials of DNA base-base pairing and mispairing. AB - An understanding of why adenine (A) pairs with thymine (T) and cytosine (C) with guanine (G) in DNA is very useful in the design of sensors and other related devices. We report the use of dissociation energies, geometries and molecular electrostatic potentials (MEPs) to justify the canonical (AT and CG) Watson-Crick pairs. We also analyze all mismatches in both configurations-cis and trans-with respect to their glycoside bonds. As expected, we found that the most stable pair configuration corresponds to CG, providing an energy criterion for that preferred configuration. The reason why A gets together with T is much more difficult to explain as the energy of this pair is smaller than the energy of some other mismatched pairs. We tested MEPs to see if they could shed light on this problem. Interestingly, MEPs yield a unique pattern (shape) for the two canonical cases but different shapes for the mismatches. A tunnel of positive potential surrounded by a negative one is found interconnecting the three H-bonds of CG and the two of AT. This MEP tunnel, assisted partially by energetics and geometrical criteria, unambiguously determine a distinctive feature of the affinity between A and T as well as that between G and C. PMID- 21625906 TI - Role of nitric oxide in amphetamine-induced sensitization of schedule-induced polydipsic rats. AB - RATIONALE: Repeated injections of amphetamine (AMPH) can progressively augment behavioral responses, a phenomenon known as behavioral sensitization. AMPH induced behavioral sensitization can be demonstrated in a rat model of schedule induced polydipsia (SIP). OBJECTIVES: The aim of this study was to investigate the effects of a nonspecific nitric oxide (NO) synthase inhibitor, N(G)-nitro arginine methyl ester (L: -NAME), on the AMPH sensitization effects in SIP. MATERIALS AND METHODS: Rats were trained to establish SIP. The induction of AMPH sensitization in SIP was tested in either the home cage or a test cage. L: -NAME (50 mg/kg, i.p.) was co-administered with AMPH for five consecutive days, and its effect on induction was examined 3 and 14 days after withdrawal. The effect of L: -NAME on expression was examined on the final testing day by co-administration with AMPH in previously sensitized SIP rats. RESULTS: AMPH-induced behavioral sensitization in SIP rats only occurred when AMPH was injected in the home cage. Pretreatment with L: -NAME successfully blocked the induction of this sensitization following both short and long-term withdrawal. Once sensitization had been established, L: -NAME had no further influence. CONCLUSIONS: These results suggest that SIP behavior observed after repeated AMPH treatment is mediated by an NO-associated mechanism. PMID- 21625907 TI - Occupancy of dopamine D2 and D3 and serotonin 5-HT1A receptors by the novel antipsychotic drug candidate, cariprazine (RGH-188), in monkey brain measured using positron emission tomography. AB - RATIONALE: Cariprazine is a novel antipsychotic drug candidate that exhibits high selectivity and affinity to dopamine D(3) and D(2) receptors and moderate affinity to serotonin 5-HT(1A) receptors. Targeting receptors other than D(2) may provide a therapeutic benefit for both positive and negative symptoms associated with schizophrenia. Positron emission tomography (PET) can be used as a tool in drug development to assess the in vivo distribution and pharmacological properties of a drug. OBJECTIVES: The objective of this study was to determine dopamine D(2)/D(3) and serotonin 5-HT(1A) receptor occupancy in monkey brain after the administration of cariprazine. METHODS: We examined three monkeys using the following PET radioligands: [(11)C]MNPA (an agonist at D(2) and D(3) receptors), [(11)C]raclopride (an antagonist at D(2) and D(3) receptors), and [(11)C]WAY-100635 (an antagonist at 5-HT(1A) receptors). During each experimental day, the first PET measurement was a baseline study, the second after a low dose of cariprazine, and the third after the administration of a high dose. RESULTS: We found that cariprazine occupied D(2)/D(3) receptors in a dose-dependent and saturable manner, with the lowest dose occupying ~5% of receptors and the highest dose showing more than 90% occupancy. 5-HT(1A) receptor occupancy was considerably lower compared with D(2)/D(3) occupancy at the same doses, with a maximal value of ~30% for the raphe nuclei. CONCLUSIONS: We conclude that cariprazine binds preferentially to dopamine D(2)/D(3) rather than to serotonin 5 HT(1A) receptors in monkey brain. These findings can be used to guide the selection of cariprazine dosing in humans. PMID- 21625908 TI - Laparoscopic sleeve gastrectomy in patients over 59 years: early recovery and 12 month follow-up. AB - BACKGROUND: Bariatric surgery has shown to be safe for patients over 60 years with good results especially considering resolving of comorbidities. Sleeve gastrectomy is considered to be safer than gastric bypass (GBP) and more effective than gastric banding with less adverse symptoms. Weight loss may be more modest than after GBP, but the effect on vitamins may also be milder. METHODS: Since 2007, we collected prospectively 12-month follow-up data from 55 sleeve gastrectomy patients of whom 12 were over 59 years of age. Vitamin and calcium supplements were used postoperatively. The recovery from the operation was recorded during hospital stay, at 1- and 12-month follow-up visits using a standard protocol including laboratory tests. The results between patients over and under 59 years were compared. RESULTS: The preoperative weight and weight loss were comparable between the groups. Operation time was shorter and hospital stay was longer for older patients, p = ns. There was no operative mortality. Early major complications were seen more often in the older age group, 42% vs 9% (p = 0.02), but late complications were more common in younger patients, 17% vs 44%, p = ns. Early complications were mostly bleedings, which did not lengthen the hospital stay, neither were re-operations nor endoscopic procedures needed. Excess weight loss and resolving of comorbidities after 12 months was comparable between the groups. However, vitamin deficiencies and hypoalbuminemia were more common in the older age group, 42% and 23% for vitamins and 44% and 29% for proteins, p = ns. The older patients had more adverse effects related to surgery, 25% vs 9%, and younger had more adverse psychiatric effects, p = ns. CONCLUSIONS: Sleeve gastrectomy is effective and safe for older bariatric patients. Weight loss is comparable to younger patients and enough to resolve the comorbidities in most of the patients. With standardized nutritional supplementation, the older patients had more often vitamin deficiencies and hypoalbuminemia. Although operative treatment of older bariatric patients is safe, their postoperative care is demanding considering vitamins and protein. PMID- 21625909 TI - Influence of activity levels and energy intake on percent excess weight loss after Roux-en-Y gastric bypass. AB - BACKGROUND: Gastric bypass is a successful medical intervention for weight loss for obesity. Weight loss is substantial after this surgery. Predictors of the most successful weight loss are not yet fully known. The purpose of this study was to define variables that improve percent excess weight loss (%EWL) in this post-surgical population. METHODS: All patients who underwent the Roux-en-Y gastric bypass (RYGB) during the 5 years preceding the study in one surgical practice (n = 805; respondents = 265; 33%) received the Arizona Activity Frequency Questionnaire and the Arizona Food Frequency Questionnaire. Analysis through ANOVA testing to determine relationships between selected behaviors and %EWL was performed. RESULTS: Comparisons of differences in mean %EWL were analyzed using the variables of energy consumption/day (energy consumption), hours of activity/day (hours in activity), and energy expended in activity/day (energy expended). Patients with more energy expended, and hours in activity demonstrated significantly better %EWL (p = 0.05) when compared to those with less energy expended or hours in activity. Reported energy consumption did not significantly influence %EWL in this study. CONCLUSIONS: This research suggests that in this sample of post-RYGB patients, energy expended in activity, as either energy expended or hours in activity improved their %EWL over those persons not expending as much energy in activity. Increasing the hours in activity improved the maintenance of %EWL in these respondents. Energy consumption did not have a statistically significant effect on %EWL or maintenance of %EWL in these subjects. PMID- 21625910 TI - Reduced cardiovascular risk following bariatric surgeries is related to a partial recovery from "adiposopathy". AB - BACKGROUND: Altered cytokine secretion from dysfunctional adipose tissue or "adiposopathy" is implicated in obesity related inflammation and may mediate reduced cardiovascular disease (CVD) risk in response to weight loss after bariatric surgery. We hypothesized that bariatric surgery reduces CVD risk by favorably altering the pro-inflammatory profile of adipose tissue as a result of weight loss. METHODS: In this observational study with repeated measures, 142 patients underwent bariatric surgery of which 45 returned for follow-up at ~6 months. At both time-points, lipid profiles and levels of plasma adiponectin, leptin, and TNF-alpha were obtained. Ratios of various adipokine parameters were related to pre- and post- surgical (gastric bypass vs. other restrictive bariatric procedures) lipid ratios. RESULTS: Prior to surgery, circulating adiponectin and the adiponectin/TNF-alpha ratio was strongly associated with CVD risk characterized by levels of triglycerides, HDL, and the TC/HDL, LDL/HDL, and TG/HDL ratios (all P < 0.05). Following bariatric surgery, BMI was decreased by 22%, adiponectin was increased by 93%, and leptin decreased by 50% as compared to baseline (all P < 0.01). TNF-alpha levels increased by 120% (P < 0.01) following surgery. Post-surgical changes in adiponectin and the leptin/adiponectin ratio were strongly associated with incremental improvements in triglycerides, HDL, and TC/HDL, LDL/HDL and TG/HDL ratios (all P < 0.05). Roux-en-y gastric bypass surgery (RYGB) as compared to other bariatric procedures was associated with more robust improvements in BMI, HDL, and leptin/adiponectin ratio than other gastric restrictive procedures (P < 0.05). CONCLUSIONS: Thus, bariatric surgery, especially RYGB, ameliorates CVD risk through a partial recovery from "adiposopathy", distinctively characterized by improved adiponectin and the leptin/adiponectin ratio. PMID- 21625912 TI - Welcome and invitation to IFSO 2011 to Hamburg and Germany. PMID- 21625911 TI - The long-term risk of venous thromboembolism following bariatric surgery. AB - BACKGROUND: Venous thromboembolism (VTE) is a leading cause of morbidity and mortality following bariatric surgery. The exact duration and magnitude of post surgery risk for VTE, however, is unclear. We analyzed a large administrative database to determine the long-term risk and predictors for VTE in patients undergoing bariatric surgery. METHODS: A private insurance claims database was used to identify 17,434 patients who underwent bariatric surgery. Longitudinal data were available for each patient for up to 12 months post-surgery. We used logistic regression to identify independent predictors for VTE events. RESULTS: The incidence of VTE during the index surgical hospitalization was .88%. This cumulative rate rose to 2.17% at 1 month and 2.99% by 6 months post-surgery. Over 74% of VTE events occurred after discharge. Risk factors identified for VTE developing by 6 months post-surgery included male sex (odds ratio (OR) = 1.68; confidence limits (CL) = 1.37-2.07), age >= 55 years (OR = 2.18; CL = 1.56-3.03), smoking (OR = 1.86; CL = 1.06-3.27), and previous VTE (OR = 7.48; CL = 5.78 9.67). The laparoscopic adjustable gastric band was less likely to result in VTE compared to open or laparoscopic gastric bypass (OR = .31; CL = .13-.75). CONCLUSIONS: The period of increased risk for VTE following bariatric surgery extends well beyond the initial hospital discharge and 30 days after surgery. The high frequency of VTE up to 6 months following bariatric surgery suggests that more aggressively extended prophylaxis should be considered in patients at higher risk for VTE. PMID- 21625913 TI - Barriers to rehabilitative care for young breast cancer survivors: a qualitative understanding. AB - PURPOSE: The goal of this study was to assess the rehabilitation needs of young women breast cancer survivors under the age of 50 and to identify factors that may impact or prevent cancer rehabilitation utilization. METHODS: Utilizing a grounded theory methodology, 35 young breast cancer survivors were interviewed twice in four Atlantic Canadian provinces. RESULTS: A considerable number of barriers exist to receiving rehabilitative care post-treatment for young breast cancer survivors. The systemic barriers include the lack of availability of services, travel issues, cost of services, and the lack of support to address the unique needs for this age group. However, the most complicated barriers to accessing rehabilitative care were personal barriers which related more to choice and circumstances, such as the lack of time due to family responsibilities and appointment fatigue. Many of these personal barriers were rooted in the complex set of gender roles of young women as patients, mothers, workers, and caregivers. CONCLUSIONS: The contexts of young women's lives can have a substantial impact on their decisions to seek and receive rehabilitative care after breast cancer treatment. The systemic barriers can be reduced by introducing more services or financial assistance; however, the personal barriers to rehabilitation services are difficult to ameliorate due to the complex set of roles within and outside the family for this group of young breast cancer survivors. Health care providers need to take into consideration the multiple contexts of women's lives when developing and promoting breast cancer rehabilitation services and programs. PMID- 21625914 TI - Mindfulness groups for men with advanced prostate cancer: a pilot study to assess feasibility and effectiveness and the role of peer support. AB - PURPOSE: Men diagnosed with advanced prostate cancer experience higher psychological distress and greater unmet supportive care needs than men with localized disease. A mindfulness-based cognitive therapy group intervention was pilot tested for acceptability and effectiveness in this patient group. METHODS: Nineteen men were initially recruited to three groups and 12 completed final assessments. Outcomes assessed included anxiety, depression, cancer-related distress, prostate cancer-specific quality of life, and mindfulness skills at baseline, immediately, and 3 months post-intervention. Satisfaction measures and in-depth interviews were undertaken post-intervention to describe men's personal experiences of the groups. RESULTS: Significant improvements were observed for anxiety (p = 0.027), avoidance (p = 0.032), and mindfulness skills (p = 0.019), with a trend for a reduction in fear of cancer recurrence (p = 0.062). Effect sizes were moderate to large. A shared group identity, acceptance of, and learning from other group members were key aspects of the group context that contributed to acceptance of progressive disease. CONCLUSIONS: Mindfulness-based group interventions appear to have utility in this patient group and show promise for reducing anxiety, avoidance, and fear of cancer recurrence. Peer learning appeared to be helpful in generating acceptance of advancing disease. PMID- 21625915 TI - Effect of boron on osteogenic differentiation of human bone marrow stromal cells. AB - Bone marrow stromal cells (BMSCs) have been well established as an ideal source of cell-based therapy for bone tissue engineering applications. Boron (B) is a notable trace element in humans; so far, the effects of boron on the osteogenic differentiation of BMSCs have not been reported. The aim of this study was to evaluate the effects of boron (0, 1, 10,100, and 1,000 ng/ml) on osteogenic differentiation of human BMSCs. In this study, BMSCs proliferation was analyzed by cell counting kit-8 (CCK8) assay, and cell osteogenic differentiation was evaluated by alkaline phosphatase (ALP) activity assay, Von Kossa staining, and real-time PCR. The results indicated that the proliferation of BMSCs was no different from the control group when added with B at the concentration of 1, 10, and 100 ng/ml respectively (P > 0.05); in contrast, 1,000 ng/ml B inhibited the proliferation of BMSCs at days 4, 7, and 14 (P < 0.05). By ALP staining, we discovered that BMSCs treated with 10 and 100 ng/ml B presented a higher ALP activity compared with control (P < 0.05). By real-time PCR, we detected the messenger RNA expression of ALP, osteocalcin, collagen type I, and bone morphogenetic proteins 7 were also increased in 10 and 100 ng/ml B treatment groups (P < 0.05). The calcium depositions were increased in 1 and 10 ng/ml B treatment groups (P < 0.05). Taken all together, it was the first time to report that B could increase osteogenic effect by stimulating osteogenic differentiation related marker gene synthesis during the proliferation and differentiation phase in human BMSCs and could be a promising approach for enhancing osteogenic capacity of cell-based construction in bone tissue engineering. PMID- 21625916 TI - Enhanced iron and zinc accumulation in genetically engineered pineapple plants using soybean ferritin gene. AB - Pineapple (Ananas comosus L. Merr., cv. "Queen") leaf bases were transformed with Agrobacterium tumefaciens strain EHA 105 harboring the pSF and pEFESF plasmids with soybean ferritin cDNA. Four to eight percent of the co-cultivated leaf bases produced multiple shoots 6 weeks after transfer to Murashige and Skoog's medium supplemented with alpha-naphthalene acetic acid 1.8 mg/l, indole-3-butyric acid 2.0 mg/l, kinetin 2.0 mg/l, cefotaxime 400 mg/l, and kanamycin 50 mg/l. Putatively transformed shoots (1-2 cm) were selected and multiplied on medium of the same composition and elongated shoots (5 cm) were rooted on liquid rooting medium supplemented with cefotaxime 400 mg/l and kanamycin 100 mg/l. The rooted plants were analyzed through PCR, genomic Southern analysis, and reverse transcription PCR. The results clearly confirmed the integration and expression of soybean ferritin gene in the transformed plants. Atomic absorption spectroscopic analysis carried out with six independently transformed lines of pSF and pEFE-SF revealed a maximum of 5.03-fold increase in iron and 2.44-fold increase in zinc accumulation in the leaves of pSF-transformed plants. In pEFE-SF transformed plants, a 3.65-fold increase in iron and 2.05-fold increase in zinc levels was observed. Few of the transgenic plants were hardened in the greenhouse and are being grown to maturity to determine the enhanced iron and zinc accumulation in the fruits. To the best of our knowledge this is the first report on the transformation of pineapple with soybean ferritin for enhanced accumulation of iron and zinc content in the transgenic plants. PMID- 21625917 TI - Investigations on the interaction between cuprous oxide nanocubes and bovine serum albumin with comprehensive spectroscopic methods. AB - The interaction between cuprous oxide (Cu(2)O) nanocubes and bovine serum albumin (BSA) was investigated from a spectroscopic angle under simulative physiological conditions. Under pH 7.4, Cu(2)O could effectively quench the intrinsic fluorescence of BSA via static quenching. The apparent binding constant (K(A)) was 3.23, 1.91, and 1.20 * 10(4) M(-1) at 298, 304, and 310 K, respectively, and the number of binding sites was 1.05. According to the Van't Hoff equation, the thermodynamic parameters (DeltaH degrees = -63.39 kJ mol(-1), DeltaS degrees = -126.45 J mol(-1) K(-1)) indicated that hydrogen bonds and van der Waals forces played a major role in stabilizing the BSA-Cu(2)O complex. Besides, the average binding distance (r(0) = 2.76 nm) and the critical energy transfer distance (R(0) = 2.35 nm) between Cu(2)O and BSA were also evaluated according to Forster's non-radioactive energy transfer theory. Furthermore, UV-visible and circular dichroism results showed that the addition of Cu(2)O changed the secondary structure of BSA and led to a decrease in alpha-helix. All results showed that BSA underwent substantial conformational changes induced by Cu(2)O, which can be very helpful in the study of nanomaterials in the application of biomaterials. PMID- 21625918 TI - Effects of aluminum exposure on the adherence, chemotaxis, and phagocytosis capacity of peritoneal macrophages in rats. AB - To investigate the effects of aluminum (Al) exposure on peritoneal macrophages of Wistar rats, four groups of ten rats each were orally exposed to 0, 13, 26, and 52 mg kg(-1) Al(3+) in form of aluminum trichloride (AlCl(3)) in drinking water for 120 days. At the end of the experimental period, the Al concentration in serum, the adherence, chemotaxis, and phagocytosis capacity of peritoneal macrophages were determined. The results showed that the Al concentration in serum significantly increased in a dose-dependent manner; the adherence, chemotaxis, and phagocytosis capacity of peritoneal macrophages decreased with the increase of Al dose, and present a dose-effective relationship. Further, they were significantly lower in the high-dose groups (P < 0.01) compared with the control group. It indicates that Al was toxic to peritoneal macrophages of rats, and the adherence, chemotaxis, and phagocytosis capacity of peritoneal macrophages in rats were significantly suppressed by exposure to 52 mg kg(-1) day Al(3+). PMID- 21625919 TI - Development of high-throughput perfusion-based microbioreactor platform capable of providing tunable dynamic tensile loading to cells and its application for the study of bovine articular chondrocytes. AB - Mammalian cells are sensitive to extracellular microenvironments. In order to precisely explore the physiological responses of cells to tensile loading, a stable and well-defined culture condition is required. In this study, a high throughput perfusion-based microbioreactor platform capable of providing dynamic equibiaxial tensile loading to the cultured cells under a steady culture condition was proposed. The mechanism of generating tensile stimulation to cells is based on the pneumatically-driven deformation of an elastic polydimethylsiloxan (PDMS) membrane which exerts tensile loading to the attached cells. By modulating the magnitude and frequency of the applied pneumatic pressure, various tensile loading can be generated in a controllable manner. In this study, the microbioreactor platform was designed with the aid of the experimentally-validated finite element (FE) analysis to ensure the loading of tensile strain to cells is uniform and definable. Based on this design, the quantitative relationship between the applied pneumatic pressure and the generated tensile strain on the PDMS membrane was established via FE analysis. Results demonstrated that the proposed device was able to generate the tensile strain range (0~0.12), which covers the physiological condition that articular chondrocytes experience tensile strain under human walking condition. In this study, moreover, the effect of tensile loading on the metabolic, biosynthetic and proliferation activities of articular chondrocytes was investigated. Results disclosed that the dynamic tensile loading of 0.12 strain at 1 Hz might significantly up-regulate the synthesis of glycosaminoglycans while such stimulation was found no significant influence on the metabolic activity, the synthesis of collagen, and the proliferation of chondrocytes. Overall, this study has presented a high throughput perfusion micro cell culture device that is suitable for precisely exploring the effect of tensile loading on cell physiology. PMID- 21625920 TI - Mercury concentration in the muscle of seven fish species from Chagan Lake, Northeast China. AB - Chagan Lake is located downstream of the Second Songhua River basin in Northeast China. It is one of the top ten inland freshwater lakes, and an important aquatic farm in China. The lake has been receiving large amounts (currently at 1.5 * 10(8) m(3)/a) of water from the river since 1984. This would pose a threat to the aquatic system of the lake because the river was seriously polluted with mercury in 1970s-1980s. The current study is the first to report the total mercury concentrations in fish found in the lake. Mercury concentrations in seven fish species collected from the lake in January 2009 were determined. The related human health risk from fish consumption was also assessed. The average concentration of mercury in the fish was 18.8 MUg/kg of wet weight, ranging from 4.5 to 37.6 MUg/kg of wet weight. A large difference in the mercury concentrations among the fish species was found. The mercury concentration was found to be higher in carnivorous species and lower in omnivorous and herbivorous species. This demonstrates greater mercury bioaccumulation in fish species at higher trophic levels. Mercury concentrations in fish showed significant positive correlations with age, length, and weight. No significant relationship was found between mercury concentrations in fish and the habitat preferences. Mercury concentrations in fish from the lake were within the limits of the international and national standards of China established for mercury. According to the reference doses established by the United States Environmental Protection Agency, the maximum safe consuming quantity considering all the fish was 297.3 g/day/person, which was more than five times as much as the current quantity (50 g/day/person) consumed by the local residents. This investigation indicates that the historical pollution of the Second Songhua River has not caused mercury bioaccumulation in fish muscle tissue of Chagan Lake. The present consumption of fish from the lake in the local area does not pose a threat to human health. PMID- 21625921 TI - Evaluating the difference between the normalized difference vegetation index and net primary productivity as the indicators of vegetation vigor assessment at landscape scale. AB - Both the net primary productivity (NPP) and the normalized difference vegetation index (NDVI) are commonly used as indicators to characterize vegetation vigor, and NDVI has been used as a surrogate estimator of NPP in some cases. To evaluate the reliability of such surrogation, here we examined the quantitative difference between NPP and NDVI in their outcomes of vegetation vigor assessment at a landscape scale. Using Landsat ETM+ data and a process model, the Boreal Ecosystem Productivity Simulator, NPP distribution was mapped at a resolution of 90 m, and total NDVI during the growing season was calculated in Heihe River Basin, Northwest China in 2002. The results from a comparison between the NPP and NDVI classification maps show that there existed a substantial difference in terms of both area and spatial distribution between the assessment outcomes of these two indicators, despite that they are strongly correlated. The degree of difference can be influenced by assessment schemes, as well as the type of vegetation and ecozone. Overall, NDVI is not a good surrogate of NPP as the indicators of vegetation vigor assessment in the study area. Nonetheless, NDVI could serve as a fairish surrogate indicator under the condition that the target region has low vegetation cover and the assessment has relatively coarse classification schemes (i.e., the class number is small). It is suggested that the use of NPP and NDVI should be carefully selected in landscape assessment. Their differences need to be further evaluated across geographic areas and biomes. PMID- 21625922 TI - Assessment of background levels of trace metals in water and soil from a remote region of Himalaya. AB - Selected trace metals were estimated by atomic absorption spectrometry in the water and soil samples collected from the remote region of Himalaya. The soil samples were analysed for soluble and acid extractable fraction of trace metals. In water samples, Ca, Na, Mg and K emerged as dominant contributors, whereas, Ca, Na, K, Mg, Fe and Pb were estimated at comparatively higher levels in the water extract of the soil. In case of acid extract of the soil samples, Ca, K, Fe, Mg, Mn and Na were found at elevated concentrations. Based on mean levels of the metals, following decreasing concentration order was observed in water samples: Ca > Na > Mg > K > Pb > Co > Cu > Zn > Mn > Cr > Fe > Cd > Li, however, in the acid extract of the soil, following order was noted: Ca > K > Fe > Mg > Mn > Na > Pb > Zn > Cr > Li > Cu > Co > Cd. The correlation study revealed appreciably diverse mutual relationships of trace metals in the water and soil samples. The multivariate cluster analyses exhibited divergent apportionment of trace metals in water and soil samples. Among the trace metals, Cd, Pb, Li, Zn, Cr, Cu, Mn and Co exhibited extreme to significant anthropogenic enrichment in the soil samples, while the rest of the metals were mostly contributed by the natural processes. PMID- 21625923 TI - RETRACTED ARTICLE: Influence of local and regional sources on the observed variability in the concentration levels of fine and coarse atmospheric aerosol water-soluble species in Athens. PMID- 21625924 TI - Environmental study of two significant solid samples: gravitation dust sediment and soil. AB - In this work are presented results of the complex study of two significant solid environmental samples: gravitation dust sediments (industrial pollutants, potential source of risk elements input to soils) and soils (component of the environment, potential source of risk elements input to food web). The first phase of this study was focused on the study of the significant chemical properties (phase composition, content of organic and inorganic carbon) of the dust and soil samples. In the second phase, the fractionation analysis was used on the evaluation of the mobility of chosen risk elements (Cu, Ni, Pb, Zn) in the studied samples. The single-step extractions were applied in the order of the isolation of the element forms (fractions), with different mobilities during defined ecological conditions by utilization of the following reagents: 1 mol dm( 3) NH(4)NO(3) for isolation of the "mobile" fraction, 0.05 mol dm(-3) ethylenediaminetetraacetic acid and 0.43 mol dm(-3) CH(3)COOH for isolation of the "mobilizable" fraction, and 2 mol dm(-3) HNO(3) for isolation of all releasable forms. On the basis of the results obtained in this study, it is possible to state that different origins and positions of solid environmental samples in the environment reflect in different chemical properties of their matrix. The different properties of the sample matrix result in different mobilities of risk elements in these kinds of samples. The fractionation analysis with single-step extraction for isolation element fractions is the method most suitable for easy checking of environmental pollution and for evaluation of risk elements cycle in the environment. PMID- 21625925 TI - Indoor radon measurement and effective dose assessment of 150 apartments in Mashhad, Iran. AB - Environmental monitoring and indoor radon measurement are important for public health, to estimate the cancer risk of respiratory system and, if necessary, to suggest proper methods that reduce indoor radon level. In this research, indoor radon concentration in the air has been measured in 150 apartments in Mashhad city. The result demonstrates about 94.7% of apartments have radon concentration less than 100 Bq/m(3), taken by WHO as the action level, and 5.3% have the concentration higher than this level. As well as, annual radon dose has been assessed using the equation for annual effective dose calculation introduced by United Nations Scientific Committee on the Effects of Atomic Radiation. PMID- 21625926 TI - Accumulation and tolerance characteristics of cadmium in Chlorophytum comosum: a popular ornamental plant and potential Cd hyperaccumulator. AB - The effects on the growth, physiological indexes and the cadmium (Cd) accumulation in Chlorophytum comosum under Cd stress were examined by pot planting. The results showed that the tolerance index (TI) of C. comosum were all above 100 in soil Cd concentration of 100 mg kg(-1). The O(2)- production rate and electrical conductivity of C. comosum were significantly positively correlated to Cd adding-concentration while the MDA content increased and had significant differences with the control. The activities of SOD, CAT, and POD all rose significantly in lower Cd concentration and the Cd threshold of them were around 10, 50 and 20 mg kg(-1), respectively. The Cd in C. comosum root and aboveground part reached 1,522 and 865.5 mg kg(-1), respectively, in Cd concentration of soil up to 200 mg kg(-1). For the advantages of high tolerance, high accumulation, and high ornamental value, C. comosum may have tremendous application value in the treatment of Cd-contaminated soils. PMID- 21625927 TI - A spectrophotometric assay method for vanadium in biological and environmental samples using 2,4-dinitrophenylhydrazine with imipramine hydrochloride. AB - A simple, rapid, and sensitive method involving the interaction of 2,4 dinitrophenylhydrazine with imipramine hydrochloride in presence of vanadium (V) in sulfuric acid medium has been proposed for the determination of vanadium. The purple-colored product developed showed an absorption maximum at 560 nm and was stable for 24 h. The working curve was linear over the concentration range of 0.1 2.8 MUg ml( - 1), with sensitivity of detection of 0.0124 MUg ml( - 1). Molar absorptivity and Sandell's sensitivity were found to be 2.6 * 10(4) l/mol cm and 0.0039 MUg cm( - 1), respectively. The accuracy of the proposed method was assessed by Student's t test and variance ratio F test, and the results were on par with the reported method. The method was successfully used in the determination of V in water, human urine, soil, and plant samples, and it was free from interference by various concomitant ions. PMID- 21625928 TI - Re Panoff et al., Risk of locoregional recurrence by receptor status in breast cancer patients receiving modern systemic therapy and post-mastectomy radiation. PMID- 21625929 TI - Interaction between goserelin and tamoxifen in a prospective randomised clinical trial of adjuvant endocrine therapy in premenopausal breast cancer. AB - Ovarian ablation improves survival in premenopausal early breast cancer, but the potential added value by luteinizing hormone-releasing hormone (LHRH) agonists to tamoxifen is still not clear. The purpose of our study is to examine the efficacy of the LHRH agonist goserelin for adjuvant therapy of premenopausal breast cancer, the role of interaction between goserelin and tamoxifen and the impact of estrogen receptor (ER) content. A total of 927 patients were included in the Stockholm part of the Zoladex in Premenopausal Patients (ZIPP) trial. They were randomly allocated in a 2 * 2 factorial study design to goserelin, tamoxifen, the combination of goserelin and tamoxifen or no endocrine therapy for 2 years, with or without chemotherapy. This is formally not a preplanned subset analysis presenting the end point first event. In this Stockholm sub-study, at a median follow-up of 12.3 years, goserelin reduced the risk of first event by 32% (P = 0.005) in the absence of tamoxifen, and tamoxifen reduced the risk by 27% (P = 0.018) in the absence of goserelin. The combined goserelin and tamoxifen treatment reduced the risk by 24% (P = 0.021) compared with no endocrine treatment. In highly ER-positive tumours, there were 29% fewer events among goserelin treated (P = 0.044) and a trend towards greater risk reduction depending on the level of ER content. The greatest risk reduction from goserelin treatment was observed among those not receiving tamoxifen (HR: 0.52, P = 0.007). In conclusion, goserelin as well as tamoxifen reduces the risk of recurrence in endocrine responsive premenopausal breast cancer. Women with strongly ER-positive tumours may benefit more from goserelin treatment. The combination of goserelin and tamoxifen is not superior to either modality alone. With the limitations of a subset trial, these data have to be interpreted cautiously. PMID- 21625930 TI - Occult breast carcinoma in breast reduction specimens in European women. AB - Breast reduction is a common surgical procedure performed by plastic and oncoplastic breast surgeons. The authors report on the incidence and management of cancer and atypical hyperplasia in breast reduction specimens from one institution over a 10-year period. All patients who underwent breast reduction surgery at Northern General Hospital, Sheffield were identified from an electronic prospective database. The histopathology reports were analyzed. Case records of all patients with significant abnormalities were retrieved and examined to identify their management and follow-up. Between October 1999 and April 2010, 1,588 patients underwent breast reduction. Nine specimens showed atypical hyperplasia (0.57%). Five cancers were detected (0.31%). Four of the five patients had normal screening mammograms 1-3 years before the reduction operation. Of these cancers, four were invasive (three lobular, one ductal) (0.25%) and one was DCIS (0.06%). A lump was felt macroscopically by the pathologist in two of the four patients with invasive cancer. The patients with DCIS did not undergo further surgery, whereas those with invasive disease underwent mastectomy (three patients) and axillary nodal staging (four patients). None of the patients with normal post-reduction breast imaging had residual cancer on histology. The incidence of occult carcinoma in breast reduction specimens is low. Patients should be counseled with regards to the possible consequences preoperatively. PMID- 21625931 TI - Magnetic resonance imaging of the brain in adenylosuccinate lyase deficiency: a report of seven cases and a review of the literature. AB - Adenylosuccinate lyase (ADSL) deficiency is a rare autosomal recessive disorder of purine metabolism. Patients may present with a wide range of neurological symptoms. Head imaging abnormalities have been reported only rarely in the scientific literature and include atrophy of the cerebral cortex, corpus callosum, cerebellar vermis, lack of myelination, delayed myelination, anomalies of the white matter, and lissencephaly. The pathogenesis of abnormalities remains unknown. To further the understanding of the spectrum of brain abnormalities associated with ADSL deficiency, we examined the magnetic resonance findings in seven Polish patients with different clinical phenotypes and genotypes. Head MRI showed impaired white matter myelination with various degrees of global supra- and infratentorial white matter loss including widening of the lateral ventricles, enlargement of the subarachnoid spaces, atrophy of the cerebrum, hypoplasia of the cerebellar hemispheres and enlargement of the cisterna magna, and white matter abnormal hyperintense signal on T(2)-weighted sequences. We recommend performing a detailed analysis of urine and plasma purine metabolites in patients who have neurological findings, including developmental delay, microcephaly, autistic features, neonatal encephalopathy, and seizures especially if MRI findings such as delayed or lack of myelination, white matter abnormal signal, and atrophy of the cerebrum and/or cerebellum are also present. Greater awareness of adenylosuccinate lyase deficiency among general pediatricians, neonatologists, pediatric neurologists, and also radiologists is the key to identifying the disorder at an early stage. PMID- 21625932 TI - Life-threatening disseminated tuberculosis as a complication of TNF-alpha blockade in an adolescent. AB - Life-threatening disseminated tuberculosis developed in a 17-year-old girl who was treated with the TNF-alpha blocker adalimumab for refractory SAPHO syndrome. The patient presented to the emergency department with dyspnea and somnolence and within 2 h developed the clinical picture of a septic shock. In addition to this unusual presentation, she showed a complicated course with increasing cerebral granuloma formation in spite of adequate antimycobacterial treatment. Immune reconstitution after discontinuation of TNF blockade may contribute to this "paradoxical reaction." Possible implications for screening, diagnosis, and treatment of tuberculosis in children and adolescents receiving anti-TNF treatment are discussed. PMID- 21625933 TI - Management of ligneous conjunctivitis in a child with plasminogen deficiency. AB - Plasminogen deficiency, a rare autosomal recessive disorder, is classified as type I (hypoplasminogenemia) or type II (dysplasminogenemia). Hypoplasminogenemia is characterized by impaired wound healing while ligneous conjunctivitis (LC) is its main manifestation presenting with redness of the conjunctivae and progression to pseudomembranes' formation on the palpebral surfaces. A 4-year-old girl with LC in her left eye and impaired vision was referred to our unit. The conjunctival membranes had been already excised twice, followed by recurrences. Soon after the third recurrence, a probable diagnosis of LC was suggested, confirmed by a reduced plasminogen activity at 20% (normal values 80-120%). Both of her parents have slightly reduced plasminogen levels (50-60%) without any relevant symptom. Fresh frozen plasma (FFP) was administered systemically and topically, initiating 2 days before surgical removal of pseudomembranes with electrocautery under general anaesthesia. Systemic FFP was administered for 12 days postoperatively, along with topical use; the later was continued thereafter for 3 months. No recurrence was noticed. The vision was improved. Two weeks after cessation of the topical treatment, pseudomembranes reappeared. Topical application of FFP was reinitiated soon thereafter, and the girl underwent a second operation to have the conjunctival pseudomembranes removed. The perioperative therapeutic management was as previously described. Systemic treatment was stopped at the end of the tenth day while topical application of FFP was being continued until now, 10 months postoperatively. No recurrence has been observed and the vision remains at 9/10. CONCLUSION: Since surgical excision of the conjunctival pseudomembranes alone in patients with LC does not protect from recurrences, the perioperative administration of FFP, both systemically and topically improves the outcome. Furthermore, the long-term application of topical FFP preparations seems to prevent recurrences and has a protective effect on the vision of these patients. PMID- 21625934 TI - Phenotype analysis in patients with early onset Parkinson's disease with and without parkin mutations. AB - The data regarding whether parkin genotype attributes phenotypic variation are conflicting. Since the incidence of parkin mutations is very low in patients with an age at onset (AAO) of >40 years, previous studies have unfairly compared phenotypes of two early onset Parkinson's disease (EOPD) groups with different AAOs. Thus, we compared the clinical features between patients with and without parkin mutations in EOPD with an AAO of <=40 years. Of the 124 patients with EOPD with an AAO of <=40 years who were recruited and screened for parkin mutations, 84 completed assessments for comparison of the phenotype according to parkin genotype. Fourteen of the 84 subjects carried two parkin mutations; 6, a single mutation; and 64, no mutations. Patients with two mutations had significantly younger AAOs, longer duration of PD, and more common family history than patients without parkin mutations. Otherwise, motor and nonmotor symptoms did not differ between them. Subgroup analysis of EOPD with an AAO of <=35 years revealed similar results. Phenotype of EOPD may depend on early AAOs rather than presence of parkin mutations. PMID- 21625935 TI - Structural and metabolic damage in brains of patients with SPG11-related spastic paraplegia as detected by quantitative MRI. AB - The goal of this work was to assess brain structural and metabolic abnormalities of subjects with SPG11 and their relevance to clinical disability by using quantitative magnetic resonance (MR) metrics. Autosomal recessive hereditary spastic paraplegia (AR-HSP) with thin corpus callosum and cognitive decline is a complex neurological disorder caused by mutations in the SPG11 gene in most cases. Little is known about the process leading to corticospinal and white matter degeneration. We performed conventional MRI/MR spectroscopic imaging ((1)H MRSI) examinations in 10 HSP patients carrying an SPG11 mutation and in 10 demographically matched healthy controls (HC). We measured in each subject cerebral white matter hyperintensities (WMHs), normalized global and cortical brain volumes, and (1)H-MRSI-derived central brain levels of N-acetylaspartate (NAA) and choline (Cho) normalized to creatine (Cr). Clinical disability was assessed according to patients' autonomy in walking. Conventional MRI showed WMHs in all patients. Global brain volumes were lower in patients than in HC (p < 0.001). Decreased values were diffusely found also in cortical regions (p < 0.01). On (1)H-MRSI, NAA/Cr values were lower in SPG11 patients than in HC (p = 0.002). Cho/Cr values did not differ between patients and HC. Cerebral volume decreases and NAA/Cr in the corona radiata correlated closely with increasing disability scores (p < 0.05). Quantitative MR measures propose that widespread structural and metabolic brain damage occur in SPG11 patients. The correlation of these MR metrics with measures of patients' disease severity suggests that they might represent adequate surrogate markers of disease outcome. PMID- 21625936 TI - From the stroke unit to the stroke competence center: corresponding beneficial clinical and financial effects. AB - The introduction of the diagnosis related groups (DRG) system in Germany has radically influenced the organization of in-hospital patient treatment. Case-mix index and duration of treatment in a stroke unit (SU) play a central role. Our SU started in 1998 and was gradually extended to the current "Stroke Competence Center" (SCC), with a total capacity of 29 patients. The SCC combines acute treatment, work-up and post-stroke management by the same specialized team. We aimed primarily at demonstrating the financial effects of this concept. Data from stroke patients treated in our SU/SCC between 2004 and 2009 were analyzed. We analyzed the number of treated stroke patients, number of thrombolytic treatments, the number of cases coded with procedure codes OPS 8-981.x and the ratio of OPS 8-981.0 (24-72 h on SU) to the higher remunerated OPS 8-981.1 (>72 h on SU). The number of treated patients increased by 118.3% (from 469 in 2004 to 1024 in 2009). The number of thrombolyses per year has more than quadrupled (2004: 46, 2009: 253, i.e. 25% of SU patients). The introduction of the stroke center concept lead to a great increase in the ratio of the higher rewarded OPS 8 981.1 to OPS 8-981.0 (from 1.5 in 2005 to 5.21 in 2009). Our data demonstrates that the SCC concept leads to a greater financial potential, while offering considerable medical advantages concerning more effective stroke treatment and work-up as well as improved flow of information and enhanced individual patient physician relationship. PMID- 21625937 TI - Hereditary sensory and autonomic neuropathy II due to novel mutation in the HSN2 gene in Mexican families. PMID- 21625938 TI - Pulsed steroids followed by glatiramer acetate to prevent inflammatory activity after cessation of natalizumab therapy: a prospective, 6-month observational study. AB - In this study, the tolerability and safety of treatment with pulsed steroids and glatiramer acetate and the occurrence of clinical and radiological activity after natalizumab (NTZ) cessation in multiple sclerosis (MS) patients were assessed. MS patients with NTZ were discontinued after 2 years of treatment, or if adverse events or disease progressed during NTZ. They were offered as alternative treatment 1 g methylprednisolone per month during 3 months followed by daily 20 mcg glatiramer acetate and were prospectively studied. Adverse events, occurrence of immune reconstitution inflammatory syndrome, clinical exacerbations, and gadolinium-enhancing lesions in MRI performed at 3 and 6 months after NTZ cessation were recorded. EDSS change during follow-up was also recorded. A total of 18 MS patients entered the study and were followed up for a mean of 10 months (range 6-18 months). There were no significant adverse events. At month 3, no patient had clinical or radiological disease activity. At month 6, 16.6% of patients had had a relapse and 55.5% of patients showed gadolinium-enhancing lesions in the MRI. After 6 months, 33.3% of patients had a further relapse. There was no IRIS, severe relapses, or significant difference between EDSS at NTZ discontinuation and after follow-up. The alternative treatment with monthly prednisolone followed by GA prevents the development of IRIS, but not the return to previous inflammatory activity, which occurs between 5 and 6 months after NTZ withdrawal. PMID- 21625939 TI - Involvement of ERK1/2/NF-kappaB signal transduction pathway in TF/FVIIa/PAR2 induced proliferation and migration of colon cancer cell SW620. AB - Our previous study has demonstrated that TF/FVIIa and protease-activated receptor 2 (PAR2) are closely related to the proliferation and migration of colon cancer cell line SW620. However, the detailed signaling cascades and underlying molecular mechanisms remain unclear. This study has investigated whether extracellular signal-regulated kinase 1 and 2 (ERK1/2) and nuclear factor kappaB (NF-kappaB) signaling pathways are involved in the events. The results revealed that PAR2-activating peptide (PAR2-AP) or FVIIa elicited time-dependent upregulation of ERK1/2 phosphorylation in SW620 cells, and the effect of FVIIa was significantly attenuated by anti-TF antibody. PAR2-AP or FVIIa also increased NF-kappaB (p65/RelA) levels among cell nuclear proteins and simultaneously decreased IkappaB-alpha levels in the cytoplasmic proteins. Such effects of FVIIa can be inhibited with anti-PAR2 or anti-TF antibodies. While ERK1/2 inhibitor (U0126) intervened with the regulatory effects of PAR2-AP and FVIIa on IkappaB alpha/NF-kappaB (p65/Rel) expression in the cells, NF-kappaB inhibitor (PDTC) partially blocked the enhancing effects of PAR2-AP and FVIIa on the proliferating and migratory ability of SW620 cells. Furthermore, the regulatory effects of PAR2 AP and FVIIa on expressions of certain proteins (IL-8, caspase-7, and TF) were also significantly abolished by PDTC. Collectively, the data in this study suggest that the interaction between FVIIa and TF induces PAR2 activation, thereby triggers the ERK1/2 and IkappaB-alpha/NF-kappaB signal transduction pathway to regulate the gene expression of IL-8, TF, and caspase-7, and ultimately promotes SW620 cell proliferation and migration. PMID- 21625940 TI - Active surveillance in prostate cancer: the need to standardize. AB - Active surveillance has been proposed as an option for patients with low-risk prostate cancer in order to reduce the effects caused by overdiagnosis. Delaying treatment and applying it only if there is evidence of progression requires a careful identification of these patients. Prostate-specific antigen (PSA) serum levels lower than 10 MUg/L and Gleason score lower than 7 are the main criteria used to select patients for active surveillance based on experience accumulated in the last two decades. In the selection of patients with active surveillance two points are taken into consideration: (a) Gleason score changes introduced by the Consensus Conference of 2005; (b) differences between assays in the measurement of PSA serum levels, in the selection of patients for active surveillance. Improving the accuracy of patient's selection for active surveillance requires that Gleason score reassignment must be taken into account, as well as the harmonization between PSA assays. The use of incorrect results leads to misclassification of patients, undermining the goals of active surveillance. PMID- 21625941 TI - Combination of CA125 and RECAF biomarkers for early detection of ovarian cancer. AB - Ovarian cancer can be cured in up to 90% of cases if diagnosed early. CA125, the most studied ovarian cancer biomarker, exhibits poor sensitivity for detecting early disease stages and low specificity to malignancy. RECAF, the alpha fetoprotein receptor, is a wide-spectrum oncofetal antigen with clinical potential for cancer diagnosis, screening, and monitoring. This study evaluated the performance of RECAF as a diagnostic tool and the sensitivity of a combination of RECAF and CA125 to detect early stages of ovarian cancer at a cutoff resulting in 100% specificity among healthy women. This retrospective case control study was designed to measure the serum levels of RECAF and CA125 in normal individuals (n=106) and cancer patients stages I/II (RECAF, n=32; CA125, n=35) and III/IV (RECAF, n=49; CA125, n=51). A competitive chemiluminescence assay was developed to measure the circulating RECAF. To eliminate any false positives, we classified as positive any patient with a RECAF or a CA125 value higher than their respective 100% specificity cutoff. We have shown that RECAF discriminated cancer and healthy donors better than CA125, particularly in the early stages (AUC(RECAF)=0.96 and AUC(CA125)=0.805). CA125 sensitivity was lower in the early stages than in the advance stages; RECAF sensitivity was high at all stages. A combination of CA125 and RECAF detected three out of four early-stage patients, with no false positives. In conclusion, the combination of RECAF and CA125 serum values provides the specificity and the sensitivity necessary to screen for ovarian cancer and in particular, to detect early stages of the disease. PMID- 21625942 TI - Estrogen receptor-alpha promoter methylation in sporadic basal-like breast cancer of Chinese women. AB - Basal-like breast cancer (BLBC) appears to be characterized by a relatively unfavorable prognosis and lack of a specific therapeutic target. Estrogen receptor-alpha (ERalpha) has been widely accepted as a prognostic marker and a predictor for endocrine therapy response of breast cancer. This study aimed to clarify the correlation of ERalpha methylation with the pathogenesis and clinicopathological significance of sporadic BLBC of Chinese women without a family history of the cancer. The methylation of ERalpha promoter was investigated in genomic DNA of 60 sporadic BLBC with 108 cases of non-BLBC as control by methylation-specific polymerase chain reaction. We also investigated the expression of p53, breast cancer gene (BRCA)-1, and BRCA-2 by immunohistochemistry and analyzed the correlation between ERalpha methylation and clinicopathological features of BLBC. ERalpha methylation was observed in 48 of 60 (80.0%) sporadic BLBC, which was significantly higher than in sporadic non BLBC cancer (47/108, 43.5%; chi2=20.89, p<0.01). No correlation was found between the ERalpha methylation and age and menopausal status, while it was significantly associated with lymph node metastasis, tumor stage, nuclear p53 accumulation, and BRCA-1 and BRCA-2 expression in sporadic BLBC. The ERalpha methylation status in basal-like breast cancer was significantly higher than in sporadic non-basal-like breast cancer. It was associated with the lymph node metastasis, tumor stage, p53 nuclear accumulation, and BRCA-1 and BRCA-2 expression in BLBC. It may play an important role in BLBC pathogenesis. PMID- 21625943 TI - Expression of ezrin correlates with poor prognosis of nasopharyngeal carcinoma. AB - A significant proportion of patients with nasopharyngeal carcinoma (NPC) will develop regional relapse or distant metastasis after treatment. So, to find valuable prognostic factors becomes very important. To elucidate the expression of ezrin in NPC and its correlation with patients' clinicopathologic characteristics, time to progression, and overall survival, immunohistochemistry stains for ezrin were applied to 200 tissue specimens of NPC. Ezrin expression had no significant correlation with sex, age, primary tumor stage (T), Epstein Barr virus (EBV) VCA-IgA antibody, EBV EA-IgA antibody, and EBV DNA copy except primary nodal status (N; p= .032). Increased ezrin expression was significantly related to higher lymph nodal metastatic rate (p< .004). Among ezrin (negative/weak), ezrin (moderate), and ezrin (intense) groups, there was a significant difference for median time to progression (not reached vs. 63.2 months vs. 46 months, P< .001) and median overall survival (100.7 months vs. 80.8 months vs. 70.6 months, P< .001). Ezrin expression status was an independent prognostic factor in multivariate (Cox) analysis. Ezrin, which was significantly correlated to prognosis of disease, may be involved in the tumorigenesis, progression, and invasion of NPC. Thus, our data offer the possibility of future therapeutic targets. PMID- 21625944 TI - Detection of viral DNA sequences in sporadic colorectal cancers in relation to CpG island methylation and methylator phenotype. AB - There is evidence that insertion of viral DNA into a mammalian genome can lead to alterations of methylation patterns. The aim of the present study was to examine the presence of DNA sequences of five human DNA viruses (assessed by PCR): JC polyoma virus (JCV), human adenovirus (AdV), Epstein-Barr virus (EBV), Kaposi sarcoma-associated herpesvirus (KSHV/HHV8) and human papillomavirus (HPV) in a cohort of 186 sporadic colorectal cancers (CRCs) and related these data with the methylation status of six CpG island methylator phenotype (CIMP)-specific genes (MLH1, CACNA1G, NEUROG1, IGF2, SOCS1, RUNX3) and seven cancer-related genes markers (p16, MINT1, MINT2, MINT31, EN1, SCTR and INHBB) assessed by methylation specific PCR in 186 and 134 CRC cases, respectively. The AdV, KSHV and HPV were detected in four (2%), two (1%) and zero CRC cases, respectively, and thus were excluded from further analyses. Although 19% and 9% of the CRCs were positive for EBV and JCV, respectively, no associations between virus presence and CpG island methylation were found after correction for multiple testing. Our results demonstrate that the presence of DNA sequences of JCV and EBV in CRC is unrelated to the methylation of the 13 cancer-related CpG islands and CIMP. PMID- 21625945 TI - Retinal vascular tortuosity in persons with diabetes and diabetic retinopathy. AB - AIM/HYPOTHESIS: The aim of this hypothesis was to examine the association of retinal vessel tortuosity with diabetes and diabetic retinopathy (DR). METHODS: A clinic-based study of 327 participants (224 with diabetes and 103 non-diabetic controls) aged >= 18 years. DR was graded from fundus photographs according to the modified Airlie House Classification system and categorised into mild non proliferative DR (NPDR), moderate NPDR and vision-threatening DR (VTDR). Retinal vessel tortuosity was measured from disc-centred retinal photographs. Measurements were taken, using a semi-automated computer program by a single grader, of arterioles and venules within 0.5 to 2 disc diameters away from the optic disc. RESULTS: There were 114 (44%) participants with DR. In the multivariate analysis, retinal arteriolar and venular tortuosity were increased in participants with diabetes without DR (mean difference 12.4 * 10(-5) and 13.3 * 10(-5), respectively; both p < 0.05) and in those with DR (mean difference 15.4 * 10(-5) and 15.0 * 10(-5), respectively; both p < 0.01) compared with non diabetic participants. Among participants with diabetes, increased arteriolar tortuosity was significantly associated with mild NPDR (OR 1.53, 95% CI 1.03 2.05, per SD increase in arteriolar tortuosity) and moderate NPDR (OR 1.67, 95% CI 1.10-2.55) but not VTDR (OR 0.91, 95% CI 0.54-1.54). No association with DR was found for venular tortuosity. CONCLUSIONS/INTERPRETATION: Persons with diabetes had more tortuous retinal vasculature than persons without diabetes. In persons with diabetes, increased arteriolar tortuosity was associated with mild and moderate stages of DR. This suggests that retinal vascular tortuosity might be an early indicator of microvascular damage in diabetes; thus, further investigation is indicated. PMID- 21625946 TI - Factors affecting podophyllotoxin yield in the ex situ grown Podophyllum hexandrum, an endangered alpine native of the western Himalayas. AB - This study reports an appreciable yield of podophyllotoxin (PDT) in P. hexandrum plants grown ex situ under polyhouse conditions of a temperate locale. The PDT content of below-ground parts was affected by both plant age and growth period. However, only the effect of plant age on PDT content was significant. Thus, the highest amounts of PDT were recorded in the below-ground parts of 2-year-old plants harvested during the late-growth period (LGP). High total soluble sugars in the below-ground parts during the early growth period (EGP) and the highest nitrate and nitrate reductase in the leaves of 2-year-old plants during the peak growth period (PGP) indicated higher mobilization and assimilation of starch and nitrate. Probably the surplus carbon and nitrogen gained during the PGP were diverted from aerial parts to below-ground parts during the LGP and in turn contributed to the synthesis of higher amounts of PDT. This study shows that commercial cultivation of P. hexandrum is possible under ex situ temperate conditions. PMID- 21625947 TI - Methanol extract from mycelium of endophytic fungus Rhizoctonia sp. induces antinociceptive and anti-inflammatory activities in mice. AB - The present study aimed to elucidate the antinociceptive and anti-inflammatory properties of the methanol extract from the mycelium of the endophytic fungus Rhizoctonia sp. (MEMRh) in mice. The antinociceptive activity was assessed using the abdominal constriction, hot plate, and formalin tests. The anti-inflammatory activity was assessed using a murine model of paw edema. Intraperitoneal administration of MEMRh (0.1, 1, 10 and 100 mg/kg, i.p.) produced an inhibition of acetic acid-induced writhing in mice for at least 8 h. In addition, all doses tested of the methanol extract were able to prevent thermal nociception in the hot-plate test. Furthermore, treatment with MEMRh (10 mg/kg, i.p.) inhibited both the early and late phases of formalin-induced nociception. This antinociceptive effect exhibited by MEMRh in the formalin test was reversed by the systemic administration of naloxone. MEMRh produced inhibition in a carrageenan-induced edema model at a dose of 10 mg/kg. The same extract also displayed significant activity against a histamine- or PGE(2)-induced edema model. The experimental data demonstrated that MEMRh showed remarkable anti-inflammatory and antinociceptive activities. Further studies are warranted to define and isolate the active anti-inflammatory and antinociceptive components from this endophytic fungus, which may yield effective agents for the treatment of inflammatory disorders. PMID- 21625948 TI - Free radical scavenging and hepatoprotective potential of Ficus microcarpa L. fil. bark extracts. AB - Successive extracts of Ficus microcarpa L. fil. bark (FMB) were tested for antioxidant and hepatoprotective activity against carbon tetrachloride- and paracetamol-induced hepatotoxicities in rats. The ethyl acetate extract of FMB exhibited significant antioxidant and hepatoprotective activity by reducing carbon tetrachloride- and paracetamol-induced changes in biochemical parameters as evidenced by enzymatic and histological examination. Pretreatment with ethyl acetate extract of FMB significantly shortened the duration of pentobarbitone induced necrosis in mice, indicating its hepatoprotective potential. Phytochemical studies confirmed the presence of the phenolic compound, catechin, in FMB, which may interfere with free-radical formation and may account for its significant hepatoprotective effects. The present study thus provides a scientific rationale for the traditional use of this plant in the management of liver disorders. PMID- 21625949 TI - Acoschimperoside P, 2'-acetate: a Hedgehog signaling inhibitory constituent from Vallaris glabra. AB - Complete (1)H and (13)C NMR assignments of acoschimperoside P, 2'-acetate (1) and a new cardiac glycoside (2), isolated from the leaves of Vallaris glabra, are described. Compound 1 was active in the assay for Hedgehog signaling inhibition. In further experiments, this compound showed a strong cytotoxicity against human pancreatic (PANC1) and human prostate (DU145) cancer cells. The expression of GLI related proteins (PTCH and BCL-2) in a dose-dependent manner was also inhibited by 1. PMID- 21625950 TI - Human amniotic membrane-derived mesenchymal stem cells labeled with superparamagnetic iron oxide nanoparticles: the effect on neuron-like differentiation in vitro. AB - Mesenchymal stem cells (MSCs) have the potential for self-renewal and multipotential differentiation to regenerate damaged tissues or recover functional absence in diseases. Superparamagnetic iron oxide nanoparticles (SPIONs) are used as contrast agents in magnetic resonance imaging (MRI) for labeling cells in vitro and for tracking SPION-labeled cells after transplantation in vivo. Human amniotic membrane-derived mesenchymal stem cells (hAM-dMSCs) have the capacity for neuron-like differentiation that could be used to cure central nervous system (CNS) diseases. The study investigated the impacts of cytotoxicity of SPIONs on neuron-like differentiation of hAM-dMSCs in both single (1*) and multiple (4*) SPIONs-labeled methods. hAM-dMSCs could be efficiently labeled at safe concentrations of SPIONs (<=14 MUg/ml) without significantly affecting their viability (>80% after a MTT assay), special surface antigens (CD29, CD44, CD90, CD105 through flow cytometry), and neuron-like differentiation (nestin and neuron-specific enolase through immunocytochemistry and reverse transcription polymerase chain reaction). Compared with multiple (4*) SPION-labeled methods, a single (1*) SPION-labeled method avoided multiple SPION labeled hAM-dMSCs and minimized the impact of cytotoxicity of SPIONs on neuron like differentiation of hAM-dMSCs. Under safe concentrations of SPIONs, a single (1*) SPION-labeled method provided appropriate viability for SPIONs-labeled hAM dMSCs and facilitated the MRI evaluation of hAM-dMSCs after transplantation. PMID- 21625951 TI - Comparison of glycation of glutathione S-transferase by methylglyoxal, glucose or fructose. AB - Glycation is a process closely related to the aging and pathogenesis of diabetic complications. In this process, reactive alpha-dicarbonyl compounds (e.g., methylglyoxal) cause protein modification accompanied with potential loss of their biological activity and persistence of damaged molecules in tissues. We suppose that glutathione S-transferases (GSTs), a group of cytosolic biotransformation enzymes, may be modified by glycation in vivo, which would provide a rationale of its use as a model protein for studying glycation reactions. Glycation of GST by methylglyoxal, fructose, and glucose in vitro was studied. The course of protein glycation was evaluated using the following criteria: enzyme activity, formation of advanced glycation end-products using fluorescence and western blotting, amine content, protein conformation, cross linking and aggregation, and changes in molecular charge of GST. The ongoing glycation by methylglyoxal 2 mM resulted in pronounced decrease in the GST activity. It also led to the loss of 14 primary amino groups, which was accompanied by changes in protein mobility during native polyacrylamide gel electrophoresis. Formation of cross links with molecular weight of 75 kDa was observed. Obtained results can contribute to understanding of changes, which proceed in metabolism of xenobiotics during diabetes mellitus and ageing. PMID- 21625952 TI - Macrophage migration inhibitory factor: a regulator of MMP13 and inflammation in titanium particles-stimulated air pouch in vivo. AB - Macrophage migration inhibitory factor (MIF) is a significant regulator of inflammatory diseases, and local inflammation plays an important role in the aseptic loosening of failed total hip arthroplasty (THA). A high-level MIF expression was found in the interfacial membrane around implants. However, the cause of increased MIF expression and the action of MIF in the process of aseptic loosening implant is still unknown. This study is to investigate MIF expression and its upregulating effect on matrix metalloproteinases (MMPs) expression in the particles-stimulated air pouches in mice that appear to closely resemble the interfacial membranes. A total of 48 murine air pouches were divided into four groups, and were injected with PBS, titanium particles suspensions, titanium particles suspensions with neutralizing antibody of MIF, and titanium particles suspensions with normal IgG, respectively. Histological and cytokine responses were evaluated. The inflammatory reaction of air pouch membranes induced by titanium particles was significantly suppressed by neutralizing antibody. The levels of MIF protein and mRNA were significantly increased in the titanium particles-stimulated air pouch membranes compared with the control groups. So were the levels of MMP13 protein and mRNA. However, the levels of MMP13 protein and mRNA were significantly reduced by neutralizing antibody. Our study demonstrates that titanium particles can cause the air pouch membranes to increase the expression of MIF, which upregulates the production of MMP13 and induces inflammatory reaction in vivo. The results indicate that MIF may play an important role in the process of aseptic-loosening implants after THA. PMID- 21625953 TI - Gallic acid-induced lung cancer cell death is accompanied by ROS increase and glutathione depletion. AB - Gallic acid (GA) is generally distributed in a variety of plants and foods, and its various biological effects have been reported. Here, we investigated the effects of GA and/or caspase inhibitors on Calu-6 and A549 lung cancer cells in relation to cell death and reactive oxygen species (ROS). The growths of Calu-6 and A549 cells were diminished with an IC(50) of approximately 30 and 150 MUM GA at 24 h, respectively. GA also inhibited the growth of primary human pulmonary fibroblast (HPF) cells with an IC(50) of about 300 MUM. GA induced apoptosis and/or necrosis in lung cancer cells, which was accompanied by the loss of mitochondrial membrane potential (MMP, DeltaPsi(m)). The percents of MMP (DeltaPsi(m)) loss and death cells by GA were lower in A549 cells than in Calu-6 cells. Caspase inhibitors did not significantly rescued lung cancer cells from GA induced cell death. GA increased ROS levels including O(2) (*-) and induced GSH depletion in both lung cancer cells. Z-VAD (pan-caspase inhibitor) did not decrease ROS levels and GSH depleted cell number in GA-treated lung cancer cells. In conclusion, GA inhibited the growth of lung cancer and normal cells. GA induced lung cancer cell death was accompanied by ROS increase and GSH depletion. PMID- 21625954 TI - HFE, MTHFR, and FGFR4 genes polymorphisms and breast cancer in Brazilian women. AB - Genetic factors related to cancer have been extensively studied and several polymorphisms have been associated to breast cancer. The FGFR4, MTHFR, and HFE genes have been associated with neoplastic diseases development. The current report outlines the analysis of the polymorphisms G388A (FGFR4), C677T (MTHFR), C282Y, and H63D (HFE) in Brazilian breast cancer patients. We studied 68 patients with invasive ductal and operable breast carcinoma and 85 women as a control group. The polymorphism frequencies in the breast cancer and control groups were analyzed, but no significant difference was observed by comparing the two groups. The presence of each polymorphism was analyzed according to the clinical features and markers already established as prognostic in the breast cancer group. The C677T, H63D, and G388A polymorphisms were not associated to histological grade, age of diagnosis, expression of HER2 receptor, or estrogen and progesterone receptor. The H63D polymorphism showed a significant association (P = 0.02) with the presence of p53 mutations, and C667T showed association to lymph node involvement (P = 0.05). Lymph node involvement, G388A polymorphism, and histological grade were independently associated to metastasis/death. Our data suggests that the polymorphisms G388A, C677T, and H63D are not useful in breast cancer diagnosis, but they may be significant additional prognostic markers related to breast cancer survival. PMID- 21625955 TI - Chronic hypoxia-induced alterations of key enzymes of glucose oxidative metabolism in developing mouse liver are mTOR dependent. AB - Hypoxia is a potent regulator of gene expression and cellular energy metabolism and known to interfere with post-natal growth and development. Although hypoxia can induce adaptive changes in the developing liver, the mechanisms underlying these changes are poorly understood. To elucidate some of the adaptive changes chronic hypoxia induces in the developing liver, we studied the expression of the genes of mammalian target of rapamycin (mTOR) signaling and glucose metabolism, undertook proteomic examination with 2D gel-MS/MS of electron transport chain, and determined activities and protein expression of several key regulatory enzymes of glucose oxidative metabolism. To gain insight into the molecular mechanism underlying hypoxia-induced liver metabolic adaptation, we treated a subset of mice with rapamycin (0.5 mg/kg/day) to inhibit mTOR postnatally. Rapamycin-treated mice showed lower birth weight, lower body weight, and liver growth retardation in a pattern similar to that observed in the hypoxic mice at P30. Rapamycin treatment led to differential impact on the cytoplasmic and mitochondrial pathways of glucose metabolism. Our results suggest a decrease in mTOR activity as part of the mechanisms underlying hypoxia-induced changes in the activities of glycolytic and TCA cycle enzymes in liver. Chronic postnatal hypoxia induces mTOR-dependent differential effects on liver glycolytic and TCA cycle enzymes and as such should be studied further as they have pathophysiological implications in hepatic diseases and conditions in which hypoxia plays a role. PMID- 21625956 TI - Eryptosis and oxidative damage in type 2 diabetic mellitus patients with chronic kidney disease. AB - It has been suggested that oxidative stress may participate in the progression of diabetes and its complications. Long-term complications of type 2 diabetes mellitus (T2DM) include retinopathy, atherosclerosis, shortened life span of erythrocytes, nephropathy, and chronic kidney disease (CKD). Oxidative damage has been associated with erythrocyte apoptosis induction in other pathological conditions. Our aim was to study the presence of eryptosis and its possible relationship with oxidative damage in patients with T2DM without CKD (T2DM/CKD( )) and in patients with T2DM and CKD (T2DM/CKD(+)).Oxidative damage of lipids erythrocytes were increased in diabetic patients. The highest lipoperoxidation was found in T2DM/CKD(+). Likewise, the lower plasma total antioxidant capacity, GSH/GSSG ratio, and GSH in erythrocytes were found in T2DM/CKD(+) patients. A negative correlation was found between plasma total antioxidant capacity and oxidative damage. Phosphatidylserine (PS) externalization was measured in erythrocytes to evaluate eryptosis. Annexin binding in erythrocytes of T2DM/CKD(+) patients was higher than in healthy subjects and T2DM/CKD(-) patients. A positive correlation between lipoperoxidation and PS externalization in erythrocytes was found. This work showed that the erythrocytes of diabetic patients have increased oxidative damage, a reduction of antioxidant systems and more erythrocyte PS externalization. The duration of diabetes and the presence of CKD increase both oxidative damage and eryptosis. It is possible that a longer time of evolution induces an increase in erythrocyte oxidative damage and the consumption of blood antioxidant systems, adding to the osmotic stress in CKD and so contributes to an increase in PS externalization in diabetic patients. PMID- 21625957 TI - Role of FAT/CD36 in novel PKC isoform activation in heart of spontaneously hypertensive rats. AB - Disruption to the sensitive balance of long-chain fatty acids and glucose in the heart could cause cardiovascular diseases. Searching for a possible role of novel protein kinase C (nPKC) in heart with disrupted energy balance, we compared the insulin-resistant spontaneously hypertensive rats (SHR), which carry a nonfunctional variant of the fatty acid transporter FAT/CD36, with the less insulin-resistant congenic strain SHR-4 that is genetically identical except for a segment on chromosome 4 including a wild-type gene for a functional FAT/CD36. We analyzed expression of the nPKC-delta and -epsilon isoforms plus triacylglycerols (TAG) content in the myocardium of both FAT/CD36 strains and after a high sucrose diet (HSD). Two weeks before killing, males of both strains were randomly divided into two groups and fed either a standard laboratory chow or an HSD. PKC was determined by Western blotting in particulate and cytosolic fractions from left ventricles. The SHR-4 rats exhibited lower serum levels of insulin and free fatty acids than did SHR rats and higher amounts of PKC-epsilon in the heart particulate fraction. HSD caused accumulation of heart TAG in SHR but not in SHR-4. HSD increased PKC-delta and decreased PKC-epsilon expression in particulate fraction from left ventricles of SHR-4 while having no effects in SHR. These results demonstrate that reduced insulin resistance in SHR-4 rats with wild-type FAT/CD36 is associated with the insulin signaling pathway involving nPKCs. PMID- 21625958 TI - Stress-induced alternations in CuZnSOD and MnSOD activity in cellular compartments of rat liver. AB - Exposure to different stressors initiates generation of reactive oxygen species (ROS), which create harmful environment for cellular macromolecules. Superoxide dismutases (SODs) represent the first line of antioxidant defense. Hence, any alternation in their function might be potentially damaging. To better define the role of SODs, we investigated the CuZnSOD activity in cytosolic and the nuclear fraction as well as mitochondrial MnSOD activity in the liver of Wistar male rats after exposure to 2 h of acute immobilization (IM) or cold (4 degrees C) stress, 21 days of chronic social isolation (IS) or their combination (chronic stress followed by acute stress). Serum corticosterone (CORT) was monitored as an indicator of the stress response. Acute IM stress, with elevated CORT level, led to increased hepatic CuZnSOD activity in the nuclear fraction. Chronic isolation stress, where CORT was close to control value, did not change the CuZnSOD activity either in nuclei or in cytosolic fraction, while combined stress IS+Cold led to increased cytosolic CuZnSOD activity. MnSOD activity in mitochondrial fraction was decreased in all treated groups. Data have shown that different stressors have diverse effect on hepatic CuZnSOD and MnSOD activity as well as on serum CORT level. Increased nuclear CuZnSOD activity after acute stress represents physiological response since the named activity protects cells against oxidative stress, while chronic IS stress compromises CuZnSOD function, suggesting an inefficient defense against ROS. Observed decrease of MnSOD activities indicate inadequate elimination of ROS after acute or chronic stress, which is characteristic of the oxidative stress. PMID- 21625959 TI - hnRNP F directs formation of an exon 4 minus variant of tumor-associated NADH oxidase (ENOX2). AB - HUVEC or mouse 3T3 cells infected with SV-40 generate within 3 to 5 days post infection an ENOX2 species corresponding to the exon-4 minus splice variant of a tumor-associated NADH oxidase (ENOX2 or tNOX) expressed at the cancer cell surface. This study was to seek evidence for splicing factors that might direct formation of the exon 4 minus ENOX2 splice variant. To determine if silencing of ENOX2 exon 4 occurs because of motifs located in exon 4, transfections were performed on MCF-10A (mammary non-cancer), BT-20 (mammary cancer), and HeLa (cervical cancer) cells using a GFP minigene construct containing either a constitutively spliced exon (albumin exon 2) or the alternatively spliced ENOX2 exon 4 between the two GFP halves. Removal of exon 4 from the processed RNA of the GFP minigene construct occurred with HeLa and to a lesser extent with BT-20 but not in non-cancer MCF-10A cells. The Splicing Rainbow Program was used to identify all of the possible hnRNPs binding sites of exon 4 of ENOX2. There are 8 Exonic Splicing Silencers (ESSs) for hnRNP binding in the exon 4 sequences. Each of these sites were mutated by site-directed mutagenesis to test if any were responsible for the splicing skip. Results showed MutG75 ESS mutation changed the GFP expression which is a sign of splicing silence, while other mutations did not. As MutG75 changed the ESS binding site for hnRNP F, this result suggests that hnRNP F directs formation of the exon 4 minus variant of ENOX2. PMID- 21625960 TI - DNA immunization against amyloid beta 42 has high potential as safe therapy for Alzheimer's disease as it diminishes antigen-specific Th1 and Th17 cell proliferation. AB - The pathogenesis of Alzheimer's disease (AD) has been strongly associated with the accumulation of amyloid beta (Abeta) peptides in brain, and immunotherapy targeting Abeta provides potential for AD prevention. A clinical trial in which AD patients were immunized with Abeta42 peptide was stopped when 6% of participants showed meningoencephalitis, apparently due to an inflammatory Th1 immune response. Previously, we and other have shown that Abeta42 DNA vaccination via gene gun generates a Th2 cellular immune response, which was shown by analyses of the respective antibody isotype profiles. We also determined that in vitro T cell proliferation in response to Abeta42 peptide re-stimulation was absent in DNA Abeta42 trimer-immunized mice when compared to Abeta42 peptide immunized mice. To further characterize this observation prospectively and longitudinally, we analyzed the immune response in wild-type mice after vaccination with Abeta42 trimer DNA and Abeta42 peptide with Quil A adjuvant. Wild-type mice were immunized with short-term (1-3* vaccinations) or long-term (6* vacinations) immunization strategies. Antibody titers and isotype profiles of the Abeta42 specific antibodies, as well as cytokine profiles and cell proliferation studies from this longitudinal study were determined. Sufficient antibody titers to effectively reduce Abeta42, but an absent T cell proliferative response and no IFNgamma or IL-17 secretion after Abeta42 DNA trimer immunization minimizes the risk of inflammatory activities of the immune system towards the self antigen Abeta42 in brain. Therefore, Abeta42 DNA trimer immunization has a high probability to be effective and safe to treat patients with early AD. PMID- 21625961 TI - Neuronal functions associated with endo- and exocytotic events-cum-molecular trafficking may be cell maturation-dependent: lessons learned from studies on botulism. AB - The passion in the scientific endeavors of Marshall Warren Nirenberg had been his quest for knowledge regarding the storage, retrieval, and processing of information in the cell. After deciphering the genetic code for which he shared the Nobel Prize in Physiology and Medicine in 1968, Nirenberg devoted his attention to unraveling the mysteries in the most complex cellular organization in the body, i.e., the nervous system, especially those governing neuronal development, plasticity, and synaptogenesis. During the tenure of the primary author (RR) as a postdoctoral Staff Fellow in the Nirenberg laboratory in the late seventies to early eighties, he had the opportunity of working on projects related to what Nirenberg used to broadly define as the "synaptic code." The major aspects of these projects dealt with the functional macromolecules relevant to neuronal growth, organization, lineage, selectivity, stabilization, synaptogenesis, and functions such as neuroexocytosis. This author's emphasis was particularly on voltage-gated calcium channels that regulate stimulus-induced neurotransmitter release. One central as well as crucial theme in these studies was the fact that the neurons had to be mature and differentiated in order to study these entities (Science 222: 794-799, 1983; Cold Spring Harb Symp Quant Biol 48: 707-715, 1983). In this communication, we illustrate how did this basic knowledge, i.e., cell maturation-dependent properties being essential for neuronal functions, led to a successful experimental design and demonstration of the validity of the targeted neurologic therapeutic delivery approach based on recombinant botulinum toxin serotype A (BoNT/A) heavy chain (rHC) serving as a neuron-specific targeting molecule (BMC Pharmacol 9: 12, 2009). PMID- 21625962 TI - Acute hemorrhagic rectal ulcer: an important cause of lower gastrointestinal bleeding in the critically ill patients. AB - BACKGROUND AND AIM: The occurrence of acute hemorrhagic rectal ulcer (AHRU) in patients in the intensive care unit (ICU) has not been well investigated. The aims of this study were to evaluate the clinical manifestations and outcomes in these patients. METHOD: The patients developing significant acute lower gastrointestinal (LGI) bleeding after ICU admission from July 2002 to December 2007 were retrospectively reviewed. Bedside colonoscopy was performed within 24 h of bleeding, and those patients with bleeding from AHRU were studied. Ulcers with stigmata of recent bleeding were treated with endoscopic hemostasis, and the outcome of these patients was analyzed. RESULTS: AHRU occurred in 36 of 114 patients (31.6%) and was the most common cause of acute LGI bleeding after ICU admission. Most patients had comorbidities, such as respiratory failure, renal failure, diabetes mellitus, or atherosclerosis. Fourteen patients (38.9%) developed hypovolemic shock after the onset of bleeding. Endoscopic therapy was performed in 29 patients with 97.2% success rate for hemostasis. Fourteen patients (48.3%) developed rebleeding within 4 weeks. This was controlled by repeated endoscopic intervention. Anticoagulant use was the risk factor for rebleeding after treatment. The survival rate at 4 weeks after bleeding was 52.8%. Logistic regression analysis revealed thrombocytopenia (platelet count <150,000/mm(3)) and more than one comorbidity were independent predictors for mortality. CONCLUSIONS: AHRU is an important etiology of acute LGI bleeding in the patients with critical illness. Bedside colonoscopy is helpful for early diagnosis and treatment. The underlying comorbidities of the patients influence the outcome after bleeding. PMID- 21625963 TI - Gastric emptying and intestinal transit of various enteral feedings following severe burn injury. AB - BACKGROUND: Burn-induced delayed gastric emptying and intestinal transit limits enteral feeding/resuscitation. AIMS: To study (1) the effects of burn injury on gastric emptying and intestinal transit at different time points following enteral feeding/fluids, and (2) the effects of enteral resuscitative fluids on gastric emptying, intestinal transit, and plasma volume expansion. METHODS: Rats were randomized into sham-burn and burn groups. They were either enterally untreated or treated by a gavage of one or multiple doses of oral rehydration solution (ORS) or, Vivonex((r)), all mixed with phenol red as a marker, at different time points from 1 to 6 h after burn. Gastric emptying, intestinal transit and hematocrit values were assessed. Gastric emptying of a semi-solid methylcellulose meal served as a standard control for gastric emptying studies. RESULTS: We found that (1) burn did not alter the gastric emptying of ORS, but delayed its intestinal transit at all time points; (2) burn delayed the gastric emptying of both methylcellulose or Vivonex and the intestinal transit of Vivonex, 6 h after burn; and (3) multiple doses of ORS normalized the elevated post-burn hematocrit values. The percentage of plasma volume expansion at 6 h resulting from the multiple-dose ORS was superior to that of Vivonex by 50%. Addition of Erythromycin to Vivonex improved its gastric emptying, intestinal transit, and plasma volume expansion. CONCLUSIONS: Burn delays the gastric emptying of semi-solids, but not the ORS. Enteral electrolyte solution (ORS) and feeding (Vivonex) provided plasma volume expansion. Prokinetic drugs may be able to maximize the effectiveness of early post-burn feeding. PMID- 21625964 TI - Generalized transit delay on wireless motility capsule testing in patients with clinical suspicion of gastroparesis, small intestinal dysmotility, or slow transit constipation. AB - BACKGROUND: The prevalence of generalized transit delay and relation to symptoms in suspected gastroparesis, intestinal dysmotility, or slow transit constipation are unknown. AIMS: The aims of this study were (1) to define prevalence of generalized dysmotility using wireless motility capsules (WMC), (2) to relate to symptoms in suspected regional delay, (3) to compare results of WMC testing to conventional transit studies to quantify new diagnoses, and (4) to assess the impact of results of WMC testing on clinical decisions. METHODS: WMC transits were analyzed in 83 patients with suspected gastroparesis, intestinal dysmotility, or slow transit constipation. RESULTS: Isolated regional delays were observed in 32% (9% stomach, 5% small bowel, 18% colon). Transits were normal in 32% and showed generalized delays in 35%. Symptom profiles were similar with normal transit, isolated delayed gastric, small intestinal, and colonic transit, and generalized delay (P = NS). Compared to conventional tests, WMC showed discordance in 38% and provided new diagnoses in 53%. WMC testing influenced management in 67% (new medications 60%; modified nutritional regimens 14%; surgical referrals 6%) and eliminated needs for testing not already done including gastric scintigraphy (17%), small bowel barium transit (54%), and radioopaque colon marker tests (68%). CONCLUSIONS: WMC testing defines localized and generalized transit delays with suspected gastroparesis, intestinal dysmotility, or slow transit constipation. Symptoms do not predict the results of WMC testing. WMC findings provide new diagnoses in >50%, may be discordant with conventional tests, and can influence management by changing treatments and eliminating needs for other tests. These findings suggest potential benefits of this method in suspected dysmotility syndromes and mandate prospective investigation to further define its clinical role. PMID- 21625965 TI - Percutaneous gastrostomy tube placement to perform transgastrostomy endoscopic retrograde cholangiopancreaticography in patients with Roux-en-Y anatomy. AB - BACKGROUND: Roux-en-Y gastric bypass (RYGB) surgery is one of the most commonly performed bariatric surgeries in the United States. Patients with prior RYGB are not amenable to conventional endoscopic retrograde cholangiopancreaticography (ERCP). Surgical gastrostomy (SG) tube placement enables transgastrostomy ERCP (TG-ERCP). MATERIALS AND METHODS: Eleven patients with RYGB anatomy received open Stamm gastrostomy after which the tract was then allowed to mature for an average of 45 days before therapeutic TG-ERCP. The success rate and procedure-related complications of both gastrostomy and ERCP were assessed. RESULTS: TG-ERCP was performed on eleven patients (median age 52 years, range 37-61 years) with prior RYGB and pancreatobiliary diseases. Indications for ERCP in these patients included suspected gallstone pancreatitis (n = 4), ampullary/biliary strictures (n = 5), pancreas divisum (n = 1), and common bile duct clipping as a result of RYGB surgery (n = 1). Two individuals developed post surgical complications with stomal-related infections. TG-ERCP with therapeutic intervention was successfully performed in all patients. Intervention included stone extractions (n = 11), biliary stricture dilation (n = 11), biliary sphincterotomy (n = 11), biliary (n = 3) and pancreatic (n = 1) stent placement, ampullary biopsies (n = 3), choledochoscopy (n = 1), and pseudocyst drainage (n = 1). Complications included post-ERCP pancreatitis (n = 2), post-sphincterotomy bleeding (n = 1), gastrostomy site bleed (n = 1), and gastric perforation (n = 1). The total number of ERCP sessions for the eleven patients was 15 (1 or 2 per patient). Median follow-up was 42 days (range 7-123 days). CONCLUSION: Surgical open gastrostomy followed by TG-ERCP enables therapeutic intervention but is associated with significant complications. PMID- 21625966 TI - The impact of obesity on egg quality. AB - Obesity in women is a concern in many countries. This causes numerous health issues; however, this review focuses on the impact of obesity on women's reproduction, and in particular the oocyte. Data from infertility clinics and experimental animal models that address the effects of obesity are presented. Bidirectional communication and metabolic support from the surrounding cumulus cells are critical for oocyte development, and the impact of obesity on these cells is also addressed. Both oocyte maturation and metabolism are impaired due to obesity, negatively impacting further development. In addition to reproductive hormones, obesity induced elevations in insulin, glucose, or free fatty acids, and changes in adipokines appear to impact the developmental competence of the oocyte. The data indicate that any one of these hormones or metabolites can impair oocyte developmental competence in vivo, and the combination of all of these factors and their interactions are the subject of ongoing investigations. PMID- 21625967 TI - A meta-analysis of the impact of IVF and ICSI on major malformations after adjusting for the effect of subfertility. AB - OBJECTIVE: To estimate the effect of assisted reproductive technology (ART) on major malformation (MM) rate in ART offspring independent of the effect of subfertility on MM. DESIGN: Meta-analysis. METHODS: This meta-analysis is based on our previously published meta-analysis of observational studies evaluating the relationship between ART treatment and MM rates, as well as recent research by Zhu et al. to estimate the impact of subfertility alone on MM in subfertile couples conceiving spontaneously. RESULTS: The overall odds ratio for MM in our original meta-analysis, in which all studies used apparently inappropriate control groups of "normal" populations, was 1.29 (95% CI 1.01-1.67). Here we attempted to estimate the risk of subfertility and used this estimate to perform an adjusted meta-analysis. Zhu et al. found that about 40% of the odds of MM was due to subfertility. When we took Zhu's finding into account, the adjusted odds ratio in the meta-analysis was 1.01 (95% CI 0.82-1.23). CONCLUSIONS: Our study suggests ART does not increase the risk of MM as much as previously reported. More research is needed to quantify the underlying risk of subfertility and separate it from the risk associated with ART. Physicians who counsel subfertile couples should recognize that previous studies of MM rates in ART patients probably overestimated the risk. PMID- 21625969 TI - Epidural anesthesia for Cesarean delivery in a patient with post-traumatic cervical syringomyelia. AB - PURPOSE: To illustrate the successful management of a patient with post-traumatic syringomyelia (PTS) and chronic pain syndrome who presented for elective Cesarean delivery under epidural anesthesia. CLINICAL FINDINGS: A 30-yr-old gravida 3 para 1 woman, with a known diagnosis of cervical PTS secondary to a whiplash injury sustained three years earlier, presented to the labour and delivery unit at 31 weeks' gestation. She had severe pain in the cervical and lumbar spine, motor and sensory deficits in the upper extremities, tender mass in her left trapezius muscle, and history of dizziness and syncopal episodes. She was taking oxycodone 120 mg.day(-1) Magnetic resonance imaging of her spine revealed a syrinx of 2 mm in diameter extending from C4 to T1 levels with disc protrusions in the C4-C6 region. There was no evidence of Arnold-Chiari malformation or elevated intracranial pressure. On airway examination, her Mallampati score appeared normal, but there was a limitation in the range of her neck movements in all directions. An elective Cesarean delivery was planned at 39 weeks' gestation. An epidural catheter was placed using ultrasound guidance, and the procedure was performed without complications. CONCLUSIONS: The successful management of this case suggests that epidural can be considered in women with cervical PTS presenting for a Cesarean delivery. PMID- 21625968 TI - Inhibition of macrophage activation and suppression of graft rejection by DTCM glutarimide, a novel piperidine derived from the antibiotic 9 methylstreptimidone. AB - OBJECTIVE: We have previously synthesized a novel piperidine compound, 3 [(dodecylthiocarbonyl)methyl]glutarimide (DTCM-glutarimide), that inhibits LPS induced NO production, and in the present research we studied further the anti inflammatory activity of DTCM-glutarimide in a macrophage cell line and in mice bearing transplanted hearts. MATERIALS AND METHODS: Mouse macrophage-like RAW264.7 cells were employed for the evaluation of cellular inflammatory activity. DTCM-glutarimide was synthesized in our laboratory. The AP-1 activity was measured by nuclear translocation and phosphorylation. For the heart transplantation experiment, male C57BL/6 (H-2b) and BALB/c (H-2d) mice were used as donor and recipient, respectively. DTCM-glutarimide was administered intraperitoneally. RESULTS: DTCM-glutarimide inhibited the LPS-induced expression of iNOS and COX-2 in macrophages; but, unexpectedly, it did not inhibit LPS induced NF-kappaB activation. Instead, it inhibited the nuclear translocation of both c-Jun and c-Fos. It also inhibited LPS-induced c-Jun phosphorylation. Moreover, it inhibited the mixed lymphocyte reaction in primary cultures of mouse spleen cells; and furthermore, in mice it prolonged the graft survival in heart transplantation experiments. CONCLUSION: The novel piperidine compound, DTCM glutarimide, was found to be a new inhibitor of macrophage activation, inhibiting AP-1 activity. It also inhibited graft rejection in mice, and thus may be a candidate for an anti-inflammatory agent. PMID- 21625970 TI - Donation after cardiocirculatory death: back to the future? PMID- 21625971 TI - Endophytes of grapevine flowers, berries, and seeds: identification of cultivable bacteria, comparison with other plant parts, and visualization of niches of colonization. AB - Endophytic bacteria can colonize various plants and organs. However, endophytes colonizing plant reproductive organs have been rarely analyzed. In this study, endophytes colonizing flowers as well as berries and seeds of grapevine plants grown under natural conditions were investigated by cultivation as well as by fluorescence in situ hybridization. For comparison, bacteria were additionally isolated from other plant parts and the rhizosphere and characterized. Flowers, fruits, and seeds hosted various endophytic bacteria. Some taxa were specifically isolated from plant reproductive organs, whereas others were also detected in the rhizosphere, endorhiza or grape inflo/infructescence stalk at the flowering or berry harvest stage. Microscopic analysis by fluorescence in situ hybridization of resin-embedded samples confirmed the presence of the isolated taxa in plant reproductive organs and enabled us to localize them within the plant. Gammaproteobacteria (including Pseudomonas spp.) and Firmicutes (including Bacillus spp.) were visualized inside the epidermis and xylem of ovary and/or inside flower ovules. Firmicutes, mainly Bacillus spp. were additionally visualized inside berries, in the intercellular spaces of pulp cells and/or xylem of pulp, but also along some cell walls inside parts of seeds. Analysis of cultivable bacteria as well as microscopic results indicated that certain endophytic bacteria can colonize flowers, berries, or seeds. Our results also indicated that some specific taxa may not only derive from the root environment but also from other sources such as the anthosphere. PMID- 21625972 TI - Impact of high-concentrate feeding and low ruminal pH on methanogens and protozoa in the rumen of dairy cows. AB - Non-lactating dairy cattle were transitioned to a high-concentrate diet to investigate the effect of ruminal pH suppression, commonly found in dairy cattle, on the density, diversity, and community structure of rumen methanogens, as well as the density of rumen protozoa. Four ruminally cannulated cows were fed a hay diet and transitioned to a 65% grain and 35% hay diet. The cattle were maintained on an high-concentrate diet for 3 weeks before the transition back to an hay diet, which was fed for an additional 3 weeks. Rumen fluid and solids and fecal samples were obtained prior to feeding during weeks 0 (hay), 1, and 3 (high concentrate), and 4 and 6 (hay). Subacute ruminal acidosis was induced during week 1. During week 3 of the experiment, there was a significant increase in the number of protozoa present in the rumen fluid (P=0.049) and rumen solids (P=0.004), and a significant reduction in protozoa in the rumen fluid in week 6 (P=0.003). No significant effect of diet on density of rumen methanogens was found in any samples, as determined by real-time PCR. Clone libraries were constructed for weeks 0, 3, and 6, and the methanogen diversity of week 3 was found to differ from week 6. Week 3 was also found to have a significantly altered methanogen community structure, compared to the other weeks. Twenty-two unique 16S rRNA phylotypes were identified, three of which were found only during high-concentrate feeding, three were found during both phases of hay feeding, and seven were found in all three clone libraries. The genus Methanobrevibacter comprised 99% of the clones present. The rumen fluid at weeks 0, 3, and 6 of all the animals was found to contain a type A protozoal population. Ultimately, high concentrate feeding did not significantly affect the density of rumen methanogens, but did alter methanogen diversity and community structure, as well as protozoal density within the rumen of nonlactating dairy cattle. Therefore, it may be necessary to monitor the rumen methanogen and protozoal communities of dairy cattle susceptible to depressed pH when methane abatement strategies are being investigated. PMID- 21625973 TI - The potential influence of short-term environmental variability on the composition of testate amoeba communities in Sphagnum peatlands. AB - Testate amoebae are a group of moisture-sensitive, shell-producing protozoa that have been widely used as indicators of changes in mean water-table depth within oligotrophic peatlands. However, short-term environmental variability (i.e., sub annual) also probably influences community composition. The objective of this study was to assess the potential influence of short-term environmental variability on the composition of testate amoeba communities in Sphagnum dominated peatlands. Testate amoebae and environmental conditions, including hourly measurements of relative humidity within the upper centimeter of the peatland surface, were examined throughout the 2008 growing season at 72 microsites within 11 peatlands of Pennsylvania and Wisconsin, USA. Relationships among testate amoeba communities, vegetation, depth to water table, pH, and an index of short-term environmental variability (EVI), were examined using nonmetric multidimensional scaling and correlation analysis. Results suggest that EVI influences testate amoeba communities, with some taxa more abundant under highly variable conditions (e.g., Arcella discoides, Difflugia pulex, and Hyalosphenia subflava) and others more abundant when environmental conditions at the peatland surface were relatively stable (e.g., Archerella flavum and Bullinularia indica). The magnitude of environmental variability experienced at the peatland surface appears to be primarily controlled by vegetation composition and density. In particular, sites with dense Sphagnum cover had lower EVI values than sites with loose-growing Sphagnum or vegetation dominated by vascular plants and/or non-Sphagnum bryophytes. Our results suggest that more environmental information may be inferred from testate amoebae than previously recognized. Knowledge of relationships between testate amoebae and short-term environmental variability should lead to more detailed and refined environmental inferences. PMID- 21625974 TI - Genetic variation and evolutionary analysis of broad bean wilt virus 2. AB - The genetic variation and evolutionary mechanisms of broad bean wilt virus 2 (BBWV-2) were studied by nucleotide sequence analysis of four genomic regions of 30 isolates from different countries. Nucleotide diversity was high (0.198) for a plant virus. Phylogenetic and genetic structure analyses showed low population subdivision, suggesting a significant gene flow between distant geographic areas. Analysis of synonymous and nonsynonymous substitutions showed different negative selection pressures for different parts of the coding regions, but no positive selection was found. Several recombination detection methods showed that some BBWV-2 genomes might have originated from recombination or reassortment. PMID- 21625975 TI - Caspase- and p38-MAPK-dependent induction of apoptosis in A549 lung cancer cells by Newcastle disease virus. AB - Newcastle disease virus (NDV) has a potential oncolytic effect due to its ability to induce apoptosis in tumor cells. However, previous studies have indicated discrepancies regarding the apoptosis signaling pathways induced by NDV in tumor cells. Here, we show that NDV infection induces simultaneous activation of intrinsic and extrinsic death pathways in A549 human lung cancer cells. In contrast, endoplasmic reticulum (ER) stress is not activated in NDV-induced apoptosis. We demonstrate for the first time that mitogen-activated protein kinase (MAPK) pathways are activated in NDV-infected A549 cells, and p38 MAPK is involved in NDV-induced cell death. Together, our findings provide novel insights into the underlying mechanisms by which NDV induces apoptosis in tumor cells. PMID- 21625976 TI - Combinations of a host resistance gene and the CI gene of turnip mosaic virus differentially regulate symptom expression in Brassica rapa cultivars. AB - In the pathosystem of Brassica rapa and turnip mosaic virus (TuMV), the type of symptoms expressed by susceptible plants are determined by the gene combinations between the host cultivar and virus strain. In this study, we found that the resistance reaction and symptoms such as systemic lethal necrosis, leaf malformation and mosaic were differentially determined, depending on the combinations of the genotypes for a host locus or two closely linked host loci and the viral CI gene. Systemic necrosis caused by TuMV-UK1 on some B. rapa subsp. pekinensis cultivars is induced in conjunction with a recessive gene, rnt1 2 (resistance and necrosis to tumv 1-2), which is allelic or closely linked to TuMV resistance gene Rnt1-1 on chromosome R6. rnt1-2 is incompletely recessive to rnt1-3, which does not cause any necrotic responses. The genotype rnt1-2/rnt1-3 caused a mild necrosis along leaf veins of severely malformed leaves. A spontaneous mutant, TuMV-UK1 (UK1m), with the amino acid substitution V1827E in CI, broke Rnt1-1 resistance and altered the systemic necrosis and leaf malformation induced by rnt1-2. This single amino acid in the CI protein of UK1 was also associated with severe mosaic and abnormal leaf development, perhaps interacting with unknown host factors. To clarify the relationship between Rnt1-1 and TuRB01b, which was previously reported as a TuMV-UK1 resistance gene on chromosome R6, the B. rapa cultivar Tropical Delight carrying TuRB01b was inoculated with UK1m or the infectious UK1 clone with the CI V1827E mutation. Because Tropical Delight showed resistance to both mutants, Rnt1-1 might be different from TuRB01b. PMID- 21625977 TI - First report of the complete sequence of Sida golden yellow vein virus from Jamaica. AB - Begomoviruses are phytopathogens that threaten food security [18]. Sida spp. are ubiquitous weed species found in Jamaica. Sida samples were collected island wide, DNA was extracted via a modified Dellaporta method, and the viral genome was amplified using degenerate and sequence-specific primers [2, 11]. The amplicons were cloned and sequenced. Sequence analysis revealed that a DNA-A molecule isolated from a plant in Liguanea, St. Andrew, was 90.9% similar to Sida golden yellow vein virus-[United States of America:Homestead:A11], making it a strain of SiGYVV. It was named Sida golden yellow vein virus-[Jamaica:Liguanea 2:2008] (SiGYVV-[JM:Lig2:08]). The cognate DNA-B, previously unreported, was successfully cloned and was most similar to that of Malvastrum yellow mosaic Jamaica virus (MaYMJV). Phylogenetic analysis suggested that this virus was most closely related to begomoviruses that infect malvaceous hosts in Jamaica, Cuba and Florida in the United States. PMID- 21625978 TI - Human cytomegalovirus latency-associated protein LUNA is expressed during HCMV infections in vivo. AB - Human cytomegalovirus (HCMV) latency is poorly understood. We previously described a novel HCMV latency-associated transcript, UL81-82ast, coding for a protein designated LUNA (latency unique natural antigen). The aim of this study was to confirm the presence of LUNA in HCMV-seropositive donors. Standard co immunoprecipitation and ELISA assays were used to detect antibodies against the LUNA protein in the sera of HCMV-seropositive donors. Specific antibodies against LUNA were detected in all HCMV-seropositive donors but in none of the seronegative donors. These data confirm that LUNA is expressed during in vivo infections and is capable of eliciting an immune response. PMID- 21625979 TI - Nitrogen uptake and preference in a forest understory following invasion by an exotic grass. AB - Plant-soil interactions have been proposed as a causative mechanism explaining how invasive plant species impact ecosystem processes. We evaluate whether an invasive plant influences plant and soil-microbe acquisition of nitrogen to elucidate the mechanistic pathways by which invaders might alter N availability. Using a (15)N tracer, we quantify differences in nitrogen uptake and allocation in communities with and without Microstegium vimineum, a shade-tolerant, C(4) grass that is rapidly invading the understories of eastern US deciduous forests. We further investigate if plants or the microbial biomass exhibit preferences for certain nitrogen forms (glycine, nitrate, and ammonium) to gain insight into nitrogen partitioning in invaded communities. Understory native plants and M. vimineum took up similar amounts of added nitrogen but allocated it differently, with native plants allocating primarily to roots and M. vimineum allocating most nitrogen to shoots. Plant nitrogen uptake was higher in invaded communities due primarily to the increase in understory biomass when M. vimineum was present, but for the microbial biomass, nitrogen uptake did not vary with invasion status. This translated to a significant reduction (P < 0.001) in the ratio of microbial biomass to plant biomass nitrogen uptake, which suggests that, although the demand for nitrogen has intensified, microbes continue to be effective nitrogen competitors. The microbial biomass exhibited a strong preference for ammonium over glycine and nitrate, regardless of invasion status. By comparison, native plants showed no nitrogen preferences and M. vimineum preferred inorganic nitrogen species. We interpret our findings as evidence that invasion by M. vimineum leads to changes in the partitioning of nitrogen above and belowground in forest understories, and to decreases in the microbial biomass, but it does not affect the outcome of plant-microbe-nitrogen interactions, possibly due to functional shifts in the microbial community as a result of invasion. PMID- 21625980 TI - The variable effects of soil nitrogen availability and insect herbivory on aboveground and belowground plant biomass in an old-field ecosystem. AB - Nutrient availability and herbivory can regulate primary production in ecosystems, but little is known about how, or whether, they may interact with one another. Here, we investigate how nitrogen availability and insect herbivory interact to alter aboveground and belowground plant community biomass in an old field ecosystem. In 2004, we established 36 experimental plots in which we manipulated soil nitrogen (N) availability and insect abundance in a completely randomized plot design. In 2009, after 6 years of treatments, we measured aboveground biomass and assessed root production at peak growth. Overall, we found a significant effect of reduced soil N availability on aboveground biomass and belowground plant biomass production. Specifically, responses of aboveground and belowground community biomass to nutrients were driven by reductions in soil N, but not additions, indicating that soil N may not be limiting primary production in this ecosystem. Insects reduced the aboveground biomass of subdominant plant species and decreased coarse root production. We found no statistical interactions between N availability and insect herbivory for any response variable. Overall, the results of 6 years of nutrient manipulations and insect removals suggest strong bottom-up influences on total plant community productivity but more subtle effects of insect herbivores on aspects of aboveground and belowground production. PMID- 21625981 TI - Evidence for positive density-dependent emigration in butterfly metapopulations. AB - A positive effect of (meta)population density on emigration has been predicted by many theoretical models and confirmed empirically in various organisms. However, in butterflies, the most popular species for dispersal studies, the evidence for its existence has so far been equivocal, with negative relationships between density and emigration being reported more frequently. We analysed dispersal in sympatric metapopulations of two Maculinea butterflies, intensively surveyed with mark-release-recapture methods for 7 years. Dispersal parameters, derived using the virtual migration model, were assessed against butterfly densities, which fluctuated strongly over the study period. Emigration was positively correlated with density, and this effect was particularly strong at densities above carrying capacity, when emigration increased up to threefold in females and twofold in males compared with the normal levels. In turn, density had little impact on other dispersal parameters analysed. Our findings provide good evidence for positive density-dependence of emigration in butterflies. Emigrating at high densities is particularly beneficial for females, because it gives them a chance to lay part of their egg-load in less crowded patches, where offspring survival is higher due to lower intraspecific competition. Even though the rise in emigration becomes considerable at densities exceeding carrying capacity, i.e. relatively infrequently, it still has serious implications for many ecological phenomena, such as species range expansions, gene flow, and metapopulation persistence. Consequently, instead of treating emigration as a fixed trait, it is worth allowing for its density-dependence in applications such as population viability analyses, genetic models or metapopulation models. PMID- 21625982 TI - Patch size matters more than dispersal distance in a mainland-island metacommunity. AB - Micro-arthropods in moss patches have been used as a model system to investigate the effects of habitat destruction and fragmentation on population viability and ecosystem functioning. Previous assessments of the sensitivity to fragmentation and the effectiveness of mitigating landscape structures have to some extent been contradictory, one possible reason being a lack of knowledge of the realised dispersal distances of the species involved. We investigated the dispersal capabilities of oribatid mites (Acari: Oribatida) and springtails (Collembola) in an experimentally fragmented system consisting of bryophytes on a bare rock surface. We used defaunated patches that were recolonized from populated patches nearby as well as from a mainland surrounding the experimental arena, during 10 weeks in summer. We measured within-mainland, mainland-to-island, and island-to island dispersal, and found that: (1) Oribatid mites were severely dispersal limited within the time frame of the experiment, even at isolation distances of only 5 cm; (2) springtails did not show any dispersal limitation over distances as far as 300 cm; (3) despite the observed dispersal limitation, the mainland had a relatively large influence on microarthropod occurrence, even at 300 cm distance; and (4) the dispersal rates were high enough for both species sorting and-in the case of collembolans-mass effects processes to occur. Our results indicate that fragmentation can strongly influence species occurrence and abundance in natural systems that are limited by dispersal. They also show that the presence of a distant mainland can override the influence of nearby habitat patches on local diversity and abundance. PMID- 21625983 TI - The effect of food limitation on immunity factors and disease resistance in the western tent caterpillar. AB - Epizootics of nucleopolyhedrovirus characterize declines of cyclic populations of western tent caterpillars, Malacosoma pluviale californicum. In field populations, infection can be apparently lacking in one generation and high in the next. This may suggest an increase in the susceptibility to infection of larvae at peak density or the triggering of a vertically transmitted virus. Here, we test the hypothesis that reduced food availability, as may occur during population outbreaks of tent caterpillars, influences the immunocompetence of larvae and increases their susceptibility to viral infection. We compared immunity factors, hemolymph phenoloxidase and hemocyte numbers, and the susceptibility to nucleopolyhedroviral infection of fifth instar larvae that were fully or partially fed as fourth instars. To determine if maternal or transgenerational influences occurred, we also determined the susceptibility of the offspring of the treated parents to viral infection. Food limitation significantly reduced larval survival, development rate, larval and pupal mass, moth fecundity and levels of hemolymph phenoloxidase, but not the numbers of hemocytes. Neither the food-reduced larvae nor their offspring were more susceptible to viral infection and were possibly even less susceptible at intermediate viral doses. Food reduction did not activate latent or covert viral infection of larvae as might be expected as a response to stress. We conclude that reducing the food intake of fourth instar larvae to an extent that had measurable and realistic impacts on their life history characteristics was not translated into increased susceptibility to viral infection. PMID- 21625984 TI - An ecological genomic approach challenging the paradigm of differential plant responses to specialist versus generalist insect herbivores. AB - A general prediction of the specialist/generalist paradigm indicates that plant responses to insect herbivores may depend on the degree of ecological specialization of the insect attacker. However, results from a single greenhouse experiment evaluating the responses of the model plant Arabidopsis thaliana to three specialist (Plutella xylostella, Pieris rapae, and Brevicoryne brassicae) and three generalist (Trichoplusia ni, Spodoptera exigua, and Myzus persicae) insect species did not support the previous prediction. Using an ecological genomic approach, we assessed plant responses in terms of herbivore-induced changes in genome-wide gene expression, defense-related pathways, and concentrations of glucosinolates (i.e., secondary metabolites that are ubiquitously present in cruciferous plants). Our results showed that plant responses were not influenced by the degree of specialization of insect herbivores. In contrast, responses were more strongly shaped by insect taxa (i.e., aphid vs. lepidopteran species), likely due to their different feeding modes. Interestingly, similar patterns of plant responses were induced by the same insect herbivore species in terms of defense signaling (jasmonic acid pathway), aliphatic glucosinolate metabolism (at both the gene expression and phenotypic levels) and genome-wide responses. Furthermore, plant responses to insect herbivores belonging to the same taxon (i.e., four lepidopteran species) were not explained by herbivore specialization or phylogenetic history. Overall, this study suggests that different feeding modes of insect taxa as well as herbivore-specific plant responses, which may result from distinct ecological/evolutionary interactions between A. thaliana (or a close relative) and each of the lepidopteran species, may explain why observed responses deviate from those predicted by the specialist/generalist paradigm. PMID- 21625986 TI - Dimensions and latent classes of episodic mania-like symptoms in youth: an empirical enquiry. AB - The dramatic increase in diagnostic rates of bipolar disorder in children and adolescents in the USA has led to an intense interest in the phenomenology of the disorder. Here we present data from a newly-developed instrument to assess episodic mania-like symptoms in youth in a large population-based sample (N = 5326) using parent- and self-report. We found that a substantial proportion of children screened positive for having episodes of "going high" and were at an increased risk for morbidity and impairment. Using factor analysis, we identified that episodic mania-like symptoms comprised two dimensions: An under-controlled dimension that was associated with significant impairment, and a low-risk exuberant dimension. Using latent class analysis, we identified a small group of children scoring high on a range of manic symptoms and suffering from severe psychosocial impairment and morbidity. Our results carry implications for the nosology and psychosocial impairment associated with episodic mood changes in young people. PMID- 21625987 TI - Molecular cloning and expression analysis of porcine ghrelin o-acyltransferase. AB - The peptide hormone ghrelin is secreted in the stomach, with unique N octanoylation at serine 3, which is a requirement for its functionality. These functions include growth hormone release, appetite stimulation, gastrointestinal motility, glucose regulation, and cell proliferation. The enzyme responsible for ghrelin acylation was recently identified as ghrelin O-acyltransferase (GOAT). In this study, porcine GOAT was cloned and characterized. A full-length cDNA of GOAT of 2013 bp was obtained, which included a 70-bp 5' UTR, a 635-bp 3' UTR, and a 1308-bp open reading frame encoding a protein of 415 amino acids. The GOAT and ghrelin mRNAs are co-expressed in stomach, pancreas, and duodenum at high levels. GOAT was also detected in liver, lung, brain, testis, spleen, kidney, heart, muscle, lipid, and ovary. Our results provide an important basis for further research on GOAT function and the relationship between ghrelin and GOAT. PMID- 21625988 TI - Clonidine extended release in the treatment of attention-deficit/hyperactivity disorder. PMID- 21625989 TI - Changes in susceptibility signs on serial T2*-weighted single-shot echo-planar gradient-echo images in acute embolic infarction: comparison with recanalization status on 3D time-of-flight magnetic resonance angiography. AB - INTRODUCTION: The present study compares changes in susceptibility signs on follow-up single-shot echo-planar gradient-echo T2*-weighted images (GRE-EPI) with vascular status on follow-up magnetic resonance angiography (MRA) in acute embolic infarction. METHODS: Twenty consecutive patients with acute embolic infarction repeatedly underwent MR imaging including GRE-EPI and MRA using a 1.5 T MR superconducting system. All patients underwent initial MR examination within 24 h of onset and follow-up MR imaging within 1 month after onset. RESULTS: Changes in susceptibility signs on follow-up GRE-EPI were compatible with vascular status on follow-up MRA in 19 of the 20 patients. Susceptibility signs disappeared with complete recanalization in 13 patients, migrated with partial recanalization in 3, did not change together with the absence of recanalization in 2, and became extended together with the absence of recanalization in 1. Cerebral hemorrhage obscured susceptibility signs in the one remaining patient. CONCLUSION: Susceptibility signs on follow-up GRE-EPI can reflect changes in an acute embolus, such as recanalization or migration, in this study. Serial GRE-EPI in acute embolism complements the diagnostic certainty of MRA by directly detecting an embolus as a susceptibility sign. PMID- 21625990 TI - Intravenous flat-detector CT angiography in acute ischemic stroke management. AB - INTRODUCTION: In the settings of stroke, a non-invasive high-resolution imaging modality to visualize the arterial intracranial circulation in the interventional lab is a helpful mean to plan the endovascular recanalization procedure. We report our initial experience with intravenously enhanced flat-detector CT (IV FDCT) technology in the detection of obstructed intracranial arteries. METHODS: Fourteen consecutive patients elected for endovascular stroke therapy underwent IV FDCT. The scans were intravenously enhanced and acquired in accordance with the previously calculated bolus arrival time. Images were processed on a commercially available workstation for reconstructions and 3D manipulation. Occlusion level and clot length, the quality of collateral vessels, and the patency of anterior and posterior communicating arteries were assessed. RESULTS: IV FDCT was performed successfully in all the cases and allowed for clot location and length visualization, assessment of communicating arteries patency, and evaluation of vessel collateral grade. Information obtained from this technique was considered useful for patients treated by endovascular approach. Retrospective review of the images by two independent readers was considered accurate and reproducible. CONCLUSIONS: IV FDCT technology provided accurate delineation of obstructed vessel segments in acute ischemic stroke disease. It gave a significant help in the interventional strategy. This new technology available in the operating room might provide a valuable tool in emerging endovascular stroke therapy. PMID- 21625991 TI - A model for linkage analysis with apomixis. AB - Apomixis, or asexual reproduction through seeds, occurs in over 400 species of angiosperms. Although apomixis can favorably perpetuate desired genotypes through successive seed generation, it may also bring about some difficulty for linkage analysis and quantitative trait locus mapping. In this article, we explore the issue of how apomixis affects the precision and power of linkage analysis with molecular markers. We derive a statistical model for estimating the linkage between different markers when some progeny are derived from apomixis. The model was constructed within the maximum likelihood framework and implemented with the EM algorithm. A series of procedures are formulated to test the linkage of markers, the rate of apomixis, and the degree of genetic interference during meiosis. The model was examined and validated through simulation studies. The model will provide a tool for linkage mapping and evolutionary studies for plant species that undergo apomixis. PMID- 21625992 TI - Prevalence of segregation distortion in diploid alfalfa and its implications for genetics and breeding applications. AB - Segregation distortion (SD) is often observed in plant populations; its presence can affect mapping and breeding applications. To investigate the prevalence of SD in diploid alfalfa (Medicago sativa L.), we developed two unrelated segregating F(1) populations and one F(2) population. We genotyped all populations with SSR markers and assessed SD at each locus in each population. The three maps were syntenic and largely colinear with the Medicago truncatula genome sequence. We found genotypic SD for 24 and 34% of markers in the F(1) populations and 68% of markers in the F(2) population; distorted markers were identified on every linkage group. The smaller percentage of genotypic SD in the F(1) populations could be because they were non-inbred and/or due to non-fully informative markers. For the F(2) population, 60 of 90 mapped markers were distorted, and they clustered into eight segregation distortion regions (SDR). Most SDR identified in the F(1) populations were also identified in the F(2) population. Genotypic SD was primarily due to zygotic rather than allelic distortion, suggesting zygotic not gametic selection is the main cause of SD. On the F(2) linkage map, distorted markers in all SDR except two showed heterozygote excess. The severe SD in the F(2) population likely biased genetic distances among markers and possibly also marker ordering and could affect QTL mapping of agronomic traits. To reduce the effects of SD and non-fully informative markers, we suggest constructing linkage maps and conducting QTL mapping in advanced generation populations. PMID- 21625993 TI - In silico mining for simple sequence repeat loci in a pineapple expressed sequence tag database and cross-species amplification of EST-SSR markers across Bromeliaceae. AB - A collection of 5,659 expressed sequence tags (ESTs) from pineapple [Ananas comosus (L.) Merr.] was screened for simple sequence repeats (EST-SSRs) with motif lengths between 1 and 6 bp. Lower thresholds of 15, 7 and 5 repeat units were used to define microsatellites of the mono-, di-, and tri- to hexanucleotide repeat type, respectively. Based on these criteria, 696 SSRs were identified among 3,389 EST unigenes, together representing 2,840 kb. This corresponds to an average density of one SSR every 4.1 kb of non-redundant EST sequences. Dinucleotide repeats were most abundant (38.4% of all SSRs) followed by trinucleotide repeats (38.1%). Flanking primer pairs were designed for 537 EST SSR loci, and 49 of these were screened for their functionality in 12 accessions of A. comosus, 14 accessions of 5 additional Ananas species and 1 species of Pseudananas. Distinct PCR products of the expected size range were obtained with 36 primer pairs. Eighteen loci analyzed in more detail were all polymorphic in pineapple, and primer pairs flanking these loci also generated PCR products from a wide range of genera and species from six subfamilies of the Bromeliaceae. The potential to reveal polymorphism in a heterologous target species was demonstrated in Deuterocohnia brevifolia (subfamily Pitcairnioideae). PMID- 21625994 TI - Expression of staphylococcal superantigens during nasal colonization is not sufficient to induce a systemic neutralizing antibody response in humans. AB - Staphylococcus aureus carriers have high-titer serum antibodies against non enterotoxin gene cluster (egc) superantigens, whereas they lack anti-egc antibodies, suggesting different superantigen expression profiles in vivo. We measured the superantigen transcripts in S. aureus directly isolated from the nose of persistent carriers and correlated them with the superantigen neutralizing antibody response. While neutralizing serum antibodies against the staphylococcal enterotoxins A and C (SEA and SEC) were found in carriers, antibodies against the egc-encoded staphylococcal enterotoxin-like toxin O (SElO) were rare. Surprisingly, the transcription of selo was comparable to sea and sec during nasal colonization. Thus, egc superantigens are transcribed during nasal colonization, but this is not sufficient to induce a serum antibody response. PMID- 21625996 TI - Anaplastic lymphoma kinase (ALK) inhibitor response in neuroblastoma is highly correlated with ALK mutation status, ALK mRNA and protein levels. AB - BACKGROUND: In pediatric neuroblastoma (NBL), high anaplastic lymphoma kinase (ALK) levels appear to be correlated with an unfavorable prognosis, regardless of ALK mutation status. This suggests a therapeutic role for ALK inhibitors in NBL patients. We examined the correlation between levels of ALK, phosphorylated ALK (pALK) and downstream signaling proteins and response to ALK inhibition in a large panel of both ALK mutated and wild type (WT) NBL cell lines. METHODS: We measured protein levels by western blot and ALK inhibitor sensitivity (TAE684) by viability assays in 19 NBL cell lines of which 6 had a point mutation and 4 an amplification of the ALK gene. RESULTS: ALK 220 kDa (p = 0.01) and ALK 140 kDa (p = 0.03) protein levels were higher in ALK mutant than WT cell lines. Response to ALK inhibition was significantly correlated with ALK protein levels (p < 0.01). ALK mutant cell lines (n = 4) were 14,9 fold (p < 0,01) more sensitive to ALK inhibition than eight WT cell lines. CONCLUSION: NBL cell lines often express ALK at high levels and are responsive to ALK inhibitors. Mutated cell lines express ALK at higher levels, which may define their superior response to ALK inhibition. PMID- 21625997 TI - Gene expression in oligodendroglial tumors. AB - BACKGROUND: Oligodendroglial tumors with 1p/19q loss are more likely to be chemosensitive and have longer survival than those with intact 1p/19q, but not all respond to chemotherapy, warranting investigation of the biological basis of chemosensitivity. METHODS: Gene expression profiling was performed using amplified antisense RNA from 28 oligodendroglial tumors treated with chemotherapy [26 serial stereotactic biopsy, 2 resection]. Expression of differentially expressed genes was validated by real-time PCR. RESULTS: Unsupervised hierarchical clustering showed clustering of multiple samples from the same case in 14/17 cases and identified subgroups associated with tumor grade and 1p/19q status. 176 genes were differentially expressed, 164 being associated with 1p/19q loss (86% not on 1p or 19q). 94 genes differed between responders and non responders to chemotherapy; 12 were not associated with 1p/19q loss. Significant differential expression was confirmed in 11/13 selected genes. Novel genes associated with response to therapy included SSBP2, GFRA1, FAP and RASD1. IQGAP1, INA, TGIF1, NR2F2 and MYCBP were differentially expressed in oligodendroglial tumors with 1p/19q loss. CONCLUSION: Gene expression profiling using serial stereotactic biopsies indicated greater homogeneity within tumors than between tumors. Genes associated with 1p/19q status or response were identified warranting further elucidation of their role in oligodendroglial tumors. PMID- 21625998 TI - Very short-term lenalidomide treatment associated with durable resolution of anemia in a patient with myelodysplastic syndrome with chromosome 5q deletion. PMID- 21625999 TI - Hodgkin's lymphoma--long-term outcome: an experience from a tertiary care cancer center in North India. AB - Limited information is available from developing countries on long-term outcome of patients with Hodgkin's lymphoma (HL). Between January 1998 and December 2005, 262 patients (age >=15 years) underwent treatment. Patients' median age was 30 years, ranging from 15 to 72 years. Male to female ratio was 2.8:1. B symptoms were present in 64% of patients. Seventy percent of patients had stage III and IV disease. Mixed cellularity (52.3%) was the most common histology followed by nodular sclerosis (38%). ABVD chemotherapy was used in 85% of the patients, and 50% received radiotherapy as consolidation. Following treatment 92% of patients achieved complete response. Five-year freedom from treatment failure (FFTF) and overall survival rate are 78.3% and 86.6% +/- 0.02% (95% CI 80.0-93.2%), respectively. Stage at presentation, number of lymph node regions involved (>=3 vs <=2), presence of B symptoms, and serum albumin (>=40 vs <40 g/L) were important determinants of FFTF. In a subset analysis of stage I and II HL patients, presence of bulky disease and pure infradiaphragmatic disease was associated with inferior outcome. On multivariate analysis involvement of three or more number of lymph node regions was a significant predictor of inferior freedom from treatment failure survival (hazard ratio 2.2, p < 0.01). Our analysis confirms excellent outcome for patients of Hodgkin's lymphoma with results comparable to developed countries. PMID- 21626000 TI - Paroxysmal non-kinesigenic dyskinesia due to spinal cord infiltration of low grade B cell non-Hodgkin's lymphoma. PMID- 21626001 TI - [Motor neuron diseases]. AB - Motor neuron diseases (MND) are a group of neurodegenerative disorders which are present in clinical, prognostic and genetic diversity. The most common MND are amyotrophic lateral sclerosis (ALS), proximal spinal muscular atrophy (SMA) and various forms of hereditary and sporadic lower motor neuron syndromes including hereditary motor neuropathies (HMN). Familial and "sporadic" forms of ALS and lower motor neuron syndromes are known. The essential pathogenic findings in MND have emerged from molecular biological examinations of the hereditary forms of MND. In ALS, one consistent neuropathological feature is intraneuronal protein inclusions which arise from TDP-43, FUS, SOD1 or ataxin-2 aggregations. TDP-43, FUS, SOD1 and ataxin-2 are multifunctional DNA/RNA-binding proteins which are involved in transcription regulation. SMA and HMN are associated with different genes whose gene products may also be involved in RNA processing. A disturbance in the regulation of RNA possibly represents an overlapping pathophysiological characteristic in MND. The elucidation of common pathways in the cascade of motor neuron degeneration is an essential point of departure for molecular genetically defined treatment strategies both in ALS and in hereditary and sporadic lower motor neuron syndromes. PMID- 21626002 TI - Protein histidine [de]phosphorylation in insulin secretion: abnormalities in models of impaired insulin secretion. AB - In the majority of cell types, including the islet beta-cell, transduction of extracellular signals involves ligand binding to a receptor, often followed by the activation G proteins and their effector modules. The islet beta-cell is unusual in that glucose lacks an extracellular receptor. Instead, events consequent to glucose metabolism promote insulin secretion via the generation of diffusible second messengers and mobilization of calcium. A selective increase in intracellular calcium has been shown to regulate the phosphorylation status key islet proteins thereby facilitating insulin secretion. In addition to classical protein kinases [e.g., protein kinases A and C], recent studies from our laboratory have focused on the expression and function of various forms of NDPK/nm23-like histidine kinases in clonal beta-cells, normal rodent, and human islets. Further, we recently reported localization of a cytosolic protein histidine phosphatase [PHP] in INS 832/13 cells, normal rat islets, and human islets. siRNA-mediated knock down of nm23-H1 and PHP in insulin-secreting INS 832/13 cells significantly attenuated glucose-induced insulin secretion. We also observed significant alterations in the expression and function of nm23-H1/PHP in beta-cells chronically exposed to elevated levels of glucose and saturated fatty acids, such as palmitate (i.e., glucolipotoxicity). Similar changes were also noted in islets from the Goto-Kakizaki and Zucker Diabetic Fatty rats, two known models for type 2 diabetes. It is concluded that protein histidine phosphorylation-dephosphorylation cycles play novel regulatory roles in G protein mediated physiological insulin secretion and that abnormalities in this signaling axis lead to impaired insulin secretion in glucolipotoxicity and type 2 diabetes. PMID- 21626004 TI - Depressed skull fracture by a three-pin head holder: a case illustration. PMID- 21626005 TI - Diagnostic and management of pediatric brain stem abscess, a case-based update. AB - PURPOSE: Authors report their experience of a pediatric brain stem abscess with a literature review. METHODS: A 2.5-year-old girl first displayed bacterial otitis 3 months before admission. Diagnosis of brain stem abscess was delayed, despite plethoric neurological signs. She complained of right hemiparesis, headache, squint, dysphagia, and false way inhalation pneumopathy. Brain magnetic resonance imaging revealed presence of an abscess in the pons, prompting for a microsurgical pus aspiration. Postoperative course was regular with a total neurological recovery. Clinical and radiological screening showed progressive regression of the pontine lesion after 4 months, despite persistence of a mild contrast enhancement. Present case is the 18th pediatric brain stem abscess reported in literature. RESULTS: Mean age of occurrence is 8.1 +/- 4 years (range 0.75-16) without gender predominance. High morbidity is typical and essentially involves motor functions. The most common clinical signs are raised intracranial pressure, motor limbs deficit, and cranial nerve palsies. Fever was found in 10 out of 18 cases (56%). Three patients received conservative medical treatment. Three patients underwent stereotactic aspiration and 12 others underwent microsurgical aspiration or excision. No mortality was reported since the eighties. However, morbidity is still high. CONCLUSION: High clinical index of suspicion is necessary to improve functional outcome. Stereotactic-guided or microsurgical aspiration are still good therapeutic options. The rarity of pediatric brain stem abscesses and the need of optimal management make this case based update very peculiar. PMID- 21626006 TI - Emanuel Swedenborg (1688-1772): pioneer of neuroanatomy. AB - Emanuel Swedenborg is widely accredited for his religious fervor and devout dedication to his spirituality. He spearheaded the creation of what is known today as the New Jerusalem Church. However, Swedenborg also served as a prominent figure in the European Enlightenment, making noteworthy strides in the fields of mathematics and science. His acumen for science instilled in the medical world groundbreaking ideas that would forever innovate the practice of medicine. Not only did Swedenborg describe intricacies of the cerebral cortex but he also discovered the perivascular spaces, the foramen of Magendie, and the cerebrospinal fluid. He noted the importance of the pituitary gland or "arch gland" in maintaining normal neurological function. Lastly, in a period where the cortex was given no significant function, Swedenborg developed the idea of somatotopic organization, and this was almost 100 years prior to Fritsch and Hitzig. It is on the shoulders of such great pioneers as Emanuel Swedenborg that we base our current understanding of the nervous system. PMID- 21626007 TI - The first posterior fossa decompression for Chiari malformation: the contributions of Cornelis Joachimus van Houweninge Graftdijk and a review of the infancy of "Chiari decompression". AB - Over the past several decades, our understanding of the pathophysiology of hindbrain hernias has markedly increased. Additionally, with the advent of MRI, diagnosis of these entities is common. Although the history of the discovery of what are now known as the Chiari malformations is well known, publications regarding the historical surgical treatment of these is, to our knowledge, not extant. Many have attributed the first successful patient series to Gardner in the 1950s. However, and unknown to many, the first description of a hindbrain decompression was in 1930 by the Dutchman Cornelis Joachimus van Houweninge Graftdijk. This neurosurgeon also added to our understanding the pathophysiology of hindbrain herniation and its relationship to raised intracranial pressure. The present paper reviews the contributions of this early pioneer of neurosurgery. PMID- 21626008 TI - Molecular profiles of EGFR, K-ras, c-met, and FGFR in pulmonary pleomorphic carcinoma, a rare lung malignancy. AB - BACKGROUND: Pulmonary pleomorphic carcinoma (PPC) is a rare type of lung cancer characterized by the poor response to conventional chemotherapy and subsequent disappointing outcomes. Therefore, it is paramount to delineate the molecular characteristics of this disease entity. METHODS: In this study, we retrospectively examined the surgical specimens of 61 patients who underwent lung surgery. Mutational or gene amplification statuses of epidermal growth factor receptor (EGFR), k-ras, c-kit, c-met, and fibroblast growth factor receptor (FGFR) were examined using genomic DNA sequencing, real-time PCR and/or fluorescence in situ hybridization (FISH). RESULTS: The median age was 61 years, and 50 patients were men and 11 were women. In the histologic review of epithelial component, adenocarcinoma were in 44 cases (72%), squamous cell carcinoma in 15 (25%) and large cell carcinoma in 2 patients (3%). Overall, 30 cases (49%) had any molecular alterations. Nine patients (15%) possessed EGFR deletion in exon 19 (n = 8) or L858R mutations in exon 21 (n = 1), while 3 other cases having atypical EGFR mutations. Six patients (9.8%) had k-ras mutations in exon 12, and 3 had c-kit mutations. High gene copy number of c-met was found in 11 patients (18.0%) and that of FGFR was in 6 patients (9.8%). No significant relationships were identified among the occurrence and type of mutations and patient survival or any other clinicopathological variables. CONCLUSIONS: Given the diverse repertoire of mutational profiles observed in PPC samples, clinical trials based on accurate cancer-genotyping should be considered as a legitimate treatment scheme for this rare disease entity in the future. PMID- 21626009 TI - Sequence and expression divergence of the AOC gene family in soybean: insights into functional diversity for stress responses. AB - As a signaling molecule, jasmonate plays a crucial role in orchestrating plant defense responses to a variety of biotic and abiotic stresses. Allene oxide cyclase (AOC: EC5.3.99.6) catalyzes the most important step in the jasmonate biosynthesis pathway. Six AOC genes were isolated from soybean, randomly distributed on chromosomes 1, 2, 8, 13, 18 and 19. The six AOC proteins were clustered into three groups with similarity values ranging from 55 to 95%. Real time PCR revealed that the AOC genes have specific and complex expression patterns in multiple organs and under several stresses. Overexpression of GmAOC1 and GmAOC5 gene in transgenic tobacco, respectively enhanced tolerance to salinity and oxidative stresses. Such a large diversity within the AOC gene family might be an adaptive mechanism that developed during soybean genome evolution. PMID- 21626011 TI - Comparison of three concentrations of simplex lidocaine in local anesthesia for inguinal hernia mesh-repairs. AB - PURPOSE: The aim of the present randomized clinical study was to assess the efficacy of simplex lidocaine in local anesthesia for inguinal hernia mesh repairs, compare analgesia of three different concentrations of lidocaine, and explore use of lower concentrations of lidocaine in local anesthesia for inguinal hernia mesh-repairs. METHODS: A total of 102 consecutive patients undergoing inguinal hernia repairs were randomized to three groups: group A (n = 34) received solution with a lidocaine concentration of 8 mg/mL, group B (n = 34) received a lidocaine concentration of 5 mg/mL, and in group C (n = 34) the lidocaine level was reduced to 3.3 mg/mL. Intraoperative pain and pain at 24 h and 48 h postoperatively were assessed by means of a visual analogue scale. Volume and doses of lidocaine used in local anesthesia were strictly recorded. RESULTS: The efficacy of simplex lidocaine in local anesthesia for inguinal hernia mesh-repairs was excellent, no patient required conversion to general anesthesia. The mean pain scores were not significantly different among the three groups. CONCLUSIONS: The local anesthesia technique was good with lidocaine alone in local anesthesia for inguinal hernia mesh-repairs. A concentration of 3.3 mg/mL lidocaine provided similar analgesia as 5 or 8 mg/mL lidocaine. PMID- 21626010 TI - Human genetics as a tool to identify progranulin regulators. AB - Frontotemporal lobar degeneration (FTLD) is a common neurodegenerative disorder that predominantly affects individuals under the age of 65. It is known that the most common pathological subtype is FTLD with TAR DNA-binding protein 43 inclusions (FTLD-TDP). FTLD has a strong genetic component with about 50% of cases having a positive family history. Mutations identified in the progranulin gene (GRN) have been shown to cause FTLD-TDP as a result of progranulin haploinsufficiency. These findings suggest a progranulin-dependent mechanism in this pathological FTLD subtype. Thus, identifying regulators of progranulin levels is essential for new therapies and treatments for FTLD and related disorders. In this review, we discuss the role of genetic studies in identifying progranulin regulators, beginning with the discovery of pathogenic GRN mutations and additional GRN risk variants. We also cover more recent genetic advances, including the detection of variants in the transmembrane protein 106 B gene that increase FTLD-TDP risk presumably by modulating progranulin levels and the identification of a potential progranulin receptor, sortilin. This review highlights the importance of genetic studies in the context of FTLD and further emphasizes the need for future genetic and cell biology research to continue the effort in finding a cure for progranulin-related diseases. PMID- 21626012 TI - Cosmetic and functional outcome after stoma site skin closure in children. AB - PURPOSE: To evaluate the functional and cosmetic outcome of stoma closure in children after straight closure (SC) versus purse-string closure (PSC). METHODS: The patients (n = 33, age 16-159 months), operated at a median age of 6 months (1 121 months) between 2007 and 2009 in our hospital, were studied to evaluate whether the proposed superiority of the PSC technique is applicable in children. The patients were operated with SC or PSC. The most common causes of the temporary stomas were necrotizing enterocolitis, Hirschsprung's disease and anal atresia. A validated scoring-system questionnaire (patient and observer scar assessment scale),POSAS was sent to the parents containing questions concerning pain, itchiness, colour, stiffness, thickness and irregularity of the scar completed by a visual analogue scale to evaluate an overall opinion. RESULTS: 25 families (SC; n = 12, PSC;n = 13) participated. The differences between the two groups are largest, although not statistically significant, for discoloration, stiffness, thickness and irregularity, with better scores in the PSC group. There was a better total POSAS score in the PSC group whilst the VAS shows very modest differences. CONCLUSION: Our study indicates advantages of the PSC technique after stoma closure with better cosmetic and functional outcome. To be able to show statistically significant differences between PSC and SC a larger study would be useful. PMID- 21626013 TI - Contemporary pediatric splenectomy: continuing controversies. AB - PURPOSE: We undertook the current study to update the literature on pediatric splenectomy in the age of minimally invasive proficiency among pediatric surgeons. The study is designed to address specific concerns among surgeons about the suitability of the laparoscopic approach in specific situations and among hematologists about the relative benefits and risks of splenectomy in children. METHODS: Retrospective analysis of clinicopathologic data for 118 children who underwent open (OS) or laparoscopic (LS) splenectomy at an urban tertiary children's hospital from January 2000 to July 2008. RESULTS: One hundred and three cases (87%) were started as LS. Operative times were equivalent for LS and OS (P = 0.8). In the LS group, there were four conversions (3.9%) from LS to OS and five early post-operative complications (4.9%). Median length of stay was 2 days for LS and 4 days for both OS and LS converted to OS (P < 0.0001). The ten largest spleens removed by LS had greater mass (P = 0.02) and tended to have greater volume (P = 0.1) than those removed by OS. Children with hereditary spherocytosis, ITP, and hemoglobinopathy had favorable clinical outcomes, regardless of operative approach. There were no cases of overwhelming post splenectomy sepsis in this series. CONCLUSIONS: Laparoscopic splenectomy is the preferred approach for splenectomy in children with hematological diseases, with or without splenomegaly. Compared to open splenectomy, laparoscopic splenectomy has equivalent operative time and improved length of stay. Both approaches have excellent therapeutic outcomes for appropriate indications. PMID- 21626014 TI - Diagnosis of gastroesophageal reflux disease in infants. AB - Gastroesophageal reflux is commonly encountered in the infant population. Most children will outgrow their reflux but some develop pervasive disease and require medical or surgical treatment. Many tools exist for use in the workup of pediatric gastroesophageal reflux disease; however, the most effective method of diagnosis is not clear. Delineating which patients will benefit from more definitive therapy is a remarkable challenge in this group, often borrowing tools and principles from the adult patient population. Therefore, we reviewed the available literature to critically evaluate the merits and limitations of the current diagnostic modalities available for the evaluation of infantile gastroesophageal reflux. PMID- 21626015 TI - Does propofol prevent testicular ischemia-reperfusion injury due to torsion in the long term? AB - OBJECTIVES: Our aim was to investigate the long-term preventive effect of propofol on testicular ischemia-reperfusion injury in a rat model. METHODS: Twenty-four adult male Sprague-Dawley rats were randomly divided into four groups (n = 6 for each group), control, sham-operated, torsion/detorsion (T/D) and T/D + propofol. Testicular ischemia was achieved by twisting the left testis 720 degrees clockwise for 2 h. Half an hour before detorsion, 50 mg/kg propofol was given intraperitoneally to the T/D + propofol group. Ipsilateral orchiectomies to determine mean testicular weights and histopathological examination according to Johnsen's mean testicular biopsy score criteria were performed 30 days after surgical procedure in all groups. RESULTS: Mean testicular weights were 1.57 +/- 0.12 g in group I, 1.59 +/- 0.36 g in group II, 0.84 +/- 0.20 g in group III and 0.87 +/- 0.29 g in group IV. Mean testicular weights decreased significantly in the T/D groups, but no improvement in testicular weight was observed with propofol administration (p 0.9372). Similarly, the Johnsen's mean testicular biopsy scores of the T/D groups were lower than those of the control and sham operated groups, but no positive effect was determined with the administration of propofol in the T/D groups (p 0.1797). CONCLUSIONS: Our results showed that there is no apparent long-term therapeutic potential attendant on using propofol in the treatment of testicular ischemia-reperfusion injury caused by testis torsion. PMID- 21626016 TI - Toxicity of polyfluorinated and perfluorinated compounds to lettuce (Lactuca sativa) and green algae (Pseudokirchneriella subcapitata). AB - Recently, polyfluorinated and perfluorinated compounds (PFCs) have been detected in most surface waters around the world. Because some PFCs are persistent and tend to accumulate in surface waters, their potential adverse effects to aquatic organisms have received increasing attention. Nevertheless, currently available toxicity information is limited. The aim of this study was to evaluate the toxicity effects of seven PFCs on root elongation of lettuce (Lactuca sativa) and photosynthesis of green algae (Pseudokirchneriella subcapitata). It was found that the toxicity profiles of both species tested were similar and had good relations with the fluorinated carbon-chain length of the PFCs investigated. One of the compounds tested, perfluorobutanoic acid, was found to be more toxic than expected in the algae test, which may be related with acidification of the test solution. It was concluded that because short-chained PFCs are becoming the predominant PFC pollutants in surface waters, their long-term toxicity and mixture toxicity with other PFCs should be studied in greater detail. PMID- 21626017 TI - The effect of two phototherapy protocols on pain control in orthodontic procedure -a preliminary clinical study. AB - Phototherapy with low-level coherent light (laser) has been reported as an analgesic and anti-inflammatory as well as having a positive effect in tissue repair in orthodontics. However, there are few clinical studies using low-level LED therapy (non-coherent light). The aim of the present study was to analyze the pain symptoms after orthodontic tooth movement associated with and not associated with coherent and non-coherent phototherapy. Fifty-five volunteers (mean age = 24.1 +/- 8.1 years) were randomly divided into four groups: G1 (control), G2 (placebo), G3 (protocol 1: laser, InGaAlP, 660 nm, 4 J/cm(2), 0.03 W, 25 s), G4 (protocol 2: LED, GaAlAs, 640 nm with 40 nm full-bandwidth at half-maximum, 4 J/cm(2), 0.10 W, 70 s). Separators were used to induce orthodontic pain and the volunteers pain levels were scored with the visual analog scale (VAS) after the separator placement, after the therapy (placebo, laser, or LED), and after 2, 24, 48, 72, 96, and 120 h. The laser group did not have statistically significant results in the reduction of pain level compared to the LED group. The LED group had a significant reduction in pain levels between 2 and 120 h compared to the control and the laser groups. The LED therapy showed a significant reduction in pain sensitivity (an average of 56%), after the orthodontic tooth movement when compared to the control group. PMID- 21626018 TI - Triptans attenuate capsaicin-induced CREB phosphorylation within the trigeminal nucleus caudalis: a mechanism to prevent central sensitization? AB - The c-AMP-responsive element binding protein (CREB) and its phosphorylated product (P-CREB) are nuclear proteins expressed after stimulation of pain producing areas of the spinal cord. There is evidence indicating that central sensitization within dorsal horn neurons is dependent on P-CREB transcriptional regulation. The objectives of the study were to investigate the expression of P CREB in cells in rat trigeminal nucleus caudalis after noxious stimulation and to determine whether pre-treatment with specific anti-migraine agents modulate this expression. CREB and P-CREB labelling was investigated within the trigeminal caudalis by immunohistochemistry after capsaicin stimulation. Subsequently, the effect of i.v. pre-treatment with either sumatriptan (n = 5), or naratriptan (n = 7) on P-CREB expression was studied. Five animals pre-treated with i.v. normal saline were served as controls. CREB and P-CREB labelling was robust in all animal groups within Sp5C. Both naratriptan and sumatriptan decreased P-CREB expression (p = 0.0003 and 0.0013) within the Sp5C. Triptans attenuate activation of CREB within the central parts of the trigeminal system, thereby leading to potential inhibition of central sensitization. P-CREB may serve as a new marker for post-synaptic neuronal activation within Sp5C in animal models relevant to migraine. PMID- 21626019 TI - Headache during airplane travel ("airplane headache"): first case in Greece. AB - Headache related to airplane flights is rare. We describe a 37-year-old female patient with multiple intense, jabbing headache episodes over the last 3 years that occur exclusively during airplane flights. The pain manifests during take off and landing, and is located always in the left retro-orbital and frontotemporal area. It is occasionally accompanied by dizziness, but no additional symptoms occur. Pain intensity diminishes and disappears after 15-20 min. Apart from occasional dizziness, no other symptoms occur. The patient has a history of tension-type headache and polycystic ovaries. Blood tests and imaging revealed no abnormalities. Here, we present the first case in Greece. We review the current literature on this rare syndrome and discuss on possible pathophysiology and the investigation of possible co-factors such as anxiety and depression. PMID- 21626020 TI - Biochemical characterization of Magnaporthe oryzae beta-glucosidases for efficient beta-glucan hydrolysis. AB - beta-Glucosidases designated MoCel3A and MoCel3B were successfully overexpressed in Magnaporthe oryzae. MoCel3A and MoCel3B showed optimal activity at 50 degrees C and pH 5.0-5.5. MoCel3A exhibited higher activity on higher degree of polymerization (DP) oligosaccharides and on beta-1,3-linked oligosaccharides than on beta-1,4-linked oligosaccharides. Furthermore, MoCel3A could liberate glucose from polysaccharides such as laminarin, 1,3-1,4-beta-glucan, phosphoric acid swollen cellulose, and pustulan, of which laminarin was the most suitable substrate. Conversely, MoCel3B preferentially hydrolyzed lower DP oligosaccharides such as cellobiose, cellotriose, and laminaribiose. Furthermore, the synergistic effects of combining enzymes including MoCel3A and MoCel3B were investigated. Depolymerization of 1,3-1,4-beta-glucan by M. oryzae cellobiohydrolase (MoCel6A) enhanced the production of glucose by the actions of MoCel3A and MoCel3B. In these reactions, MoCel3A hydrolyzed higher DP oligosaccharides, resulting in the release of glucose and cellobiose, and MoCel3B preferentially hydrolyzed lower DP oligosaccharides including cellobiose. On the other hand, MoCel3A alone produced glucose from laminarin at levels equivalent to 80% of maximal hydrolysis obtained by the combined action of MoCel3A, MoCel3B, and endo-1,3-beta-glucanase. Therefore, MoCel3A and MoCel3B activities yield glucose from not only cellulosic materials but also hemicellulosic polysaccharides. PMID- 21626021 TI - Flavonoids biotransformation by bacterial non-heme dioxygenases, biphenyl and naphthalene dioxygenase. AB - This review details recent progresses in the flavonoid biotransformation by bacterial non-heme dioxygenases, biphenyl dioxygenase (BDO), and naphthalene dioxygenase (NDO), which can initially activate biphenyl and naphthalene with insertion of dioxygen in stereospecfic and regiospecific manners. Flavone, isoflavone, flavanone, and isoflavanol were biotransformed by BDO from Pseudomonas pseudoalcaligenes KF707 and NDO from Pseudomonas sp. strain NCIB9816 4, respectively. In general, BDO showed wide range of substrate spectrum and produced the oxidized products, whereas NDO only metabolized flat two-dimensional substrates of flavone and isoflavone. Furthermore, biotransformation of B-ring skewed substrates, flavanone and isoflavanol, by BDO produced the epoxide products, instead of dihydrodiols. These results support the idea that substrate driven reactivity alteration of the Fe-oxo active species may occur in the active site of non-heme dioxygenases. The study of flavonoid biotransformation by structurally-well defined BDO and NDO will provide the substrate structure and reactivity relationships and eventually establish the production of non-plant originated flavonoids by means of microbial biotechnology. PMID- 21626022 TI - PCR methods for the rapid detection and identification of four pathogenic Legionella spp. and two Legionella pneumophila subspecies based on the gene amplification of gyrB. AB - A total of 25 gyrB gene sequences from 20 Legionella pneumophila subsp. pneumophila strains and five L. pneumophila subsp. fraseri strains were obtained and analyzed, and a multiplex PCR for the simultaneous detection of Legionella bozemanae, Legionella longbeachae, Legionella micdadei and Legioenella pneumophila, and two single PCRs for the differentiation of L. pneumophila subsp. pneumophila and L. pneumophila subsp. fraseri were established. The multiplex PCR method was shown to be highly specific and reproducible when tested against 41 target strains and 17 strains of other bacteria species. The sensitivity of the multiplex PCR was also analyzed and was shown to detect levels as low as 1 ng of genomic DNA or 10 colony-forming units (CFUs) per milliliter in mock water samples. Sixty-three air conditioner condensed water samples from Shanghai City were examined, and the result was validated using 16S rRNA sequencing. The data reported here demonstrate that the multiplex PCR method described is efficient and convenient for the detection of Legionella species in water samples. Twenty L. pneumophila subsp. pneumophila strains and five L. pneumophila subsp. fraseri strains were used for the validation of the two L. pneumophila subspecies specific PCR methods, and the results indicated that the two PCR methods were both highly specific and convenient for the identification of L. pneumophila at the subspecies level. PMID- 21626023 TI - Production of dihydrodaidzein and dihydrogenistein by a novel oxygen-tolerant bovine rumen bacterium in the presence of atmospheric oxygen. AB - The original bovine rumen bacterial strain Niu-O16, capable of anaerobically bioconverting isoflavones daidzein and genistein to dihydrodaidzein (DHD) and dihydrogenistein (DHG), respectively, is a rod-shaped obligate anaerobic bacterium. After a long-term domestication, an oxygen-tolerant bacterium, which we named Aeroto-Niu-O16 was obtained. Strain Aeroto-Niu-O16, which can grow in the presence of atmospheric oxygen, differed from the original obligate anaerobic bacterium Niu-O16 by various characteristics, including a change in bacterial shape (from rod to filament), in biochemical traits (from indole negative to indole positive and from amylohydrolysis positive to negative), and point mutations in 16S rRNA gene (G398A and G438A). We found that strain Aeroto-Niu-O16 not only grew aerobically but also converted isoflavones daidzein and genistein to DHD and DHG in the presence of atmospheric oxygen. The bioconversion rate of daidzein and genistein by strain Aeroto-Niu-O16 was 60.3% and 74.1%, respectively. And the maximum bioconversion capacity for daidzein was 1.2 and 1.6 mM for genistein. Furthermore, when we added ascorbic acid (0.15%, m/v) in the cultural medium, the bioconversion rate of daidzein was increased from 60.3% to 71.7%, and that of genistein from 74.1% to 89.2%. This is the first reported oxygen-tolerant isoflavone biotransforming pure culture capable of both growing and executing the reductive activity under aerobic conditions. PMID- 21626024 TI - Effects of cryoprotectants on viability of Lactobacillus reuteri CICC6226. AB - Freeze-drying is commonly used to preserve probiotics, but it could cause cell damage and loss of viability. The cryoprotectants play an important role in the conservation of viability during freeze-drying. In this study, we investigated the survival rates of Lactobacillus reuteri CICC6226 in the presence of cryoprotectants such as sucrose, trehalose, and reconstituted skim milk (RSM). In addition, we determined the activities of hexokinase (HK), pyruvate kinase (PK), lactate dehydrogenase (LDH), and ATPases immediately following the freeze-drying. The results showed that the differences in HK and PK activities with and without the cryoprotectants during freeze-drying were not significant, but cell viability and activities of LDH and ATPase were significantly different (P<0.01) prior to and after freeze-drying. Meanwhile, the results showed that the maintenance of the membrane integrity and fluidity was improved in the presence of the 10% trehalose or 10% RSM than other treatments during freeze-drying. These results have provided direct biochemical and metabolic evidence of injured cell during freeze-drying. Freeze-drying damaged membrane structure and function of cell and inactivated enzymes (LDH and ATPases). The results imply that LDH and ATPases are key markers and could be used to evaluate the effect of cryoprotectants on viability and metabolic activities of L. reuteri CICC6226 during freeze-drying. PMID- 21626025 TI - Reductive dehalogenation mediated initiation of aerobic degradation of 2-chloro-4 nitrophenol (2C4NP) by Burkholderia sp. strain SJ98. AB - Burkholderia sp. strain SJ98 (DSM 23195) was previously isolated and characterized for degradation and co-metabolic transformation of a number nitroaromatic compounds. In the present study, we evaluated its metabolic activity on chlorinated nitroaromatic compounds (CNACs). Results obtained during this study revealed that strain SJ98 can degrade 2-chloro-4-nitrophenol (2C4NP) and utilize it as sole source of carbon, nitrogen, and energy under aerobic conditions. The cells of strain SJ98 removed 2C4NP from the growth medium with sequential release of nearly stoichiometric amounts of chloride and nitrite in culture supernatant. Under aerobic degradation conditions, 2C4NP was transformed into the first intermediate that was identified as p-nitrophenol by high performance liquid chromatography, LCMS-TOF, and GC-MS analyses. This transformation clearly establishes that the degradation of 2C4NP by strain SJ98 is initiated by "reductive dehalogenation"; an initiation mechanism that has not been previously reported for microbial degradation of CNAC under aerobic conditions. PMID- 21626026 TI - Myositis proliferans: diagnosis and therapy of a pseudosarcomatous soft tissue lesion. AB - Myositis proliferans is a soft tissue neoplasia with rare incidence. In the most cases, it is localized in the region of the neck, shoulder, pelvis and thigh. Due to its rapid growth and histological picture, the tumour may appear as a malignant neoplasia. We report the case of a 29-year-old woman suffering from an increasing painful swelling of the left proximal lower leg. Performed biopsies and histological examinations provided the diagnosis of myositis proliferans adjacent to the fibula, which responded to local resection and did not recur after 2 years. We show the importance of adequate diagnostic and therapeutic approach to avoid unnecessary and probably radical overtherapy of the patient. PMID- 21626027 TI - Association of CD4 enhancer gene polymorphisms with rheumatoid arthritis in Egyptian female patients. AB - CD4 is a candidate gene in autoimmune diseases, including rheumatoid arthritis (RA). Because the CD4 receptor is crucial for appropriate antigen responses of CD4+ T cells, changes in CD4 expression and CD4+ T-cell activity may influence tolerance or tissue destruction in autoimmune diseases and contribute to their risk. We analyzed two polymorphisms of the CD4 in 172 female Egyptian patients with RA and in 112 matched healthy control. Genotyping of CD4-11743 and CD4-10845 was determined by restriction fragment length polymorphism-polymerase chain reaction (PCR-RFLP). Subjects with the CC genotype of CD4-11743 were significantly more likely to develop RA (OR = 2.7, P = 0.03) and more likely to have sever RA (OR = 2.7, P = 0.024). Carrier of A allele of CD4-10845 was significantly more likely to develop sever RA (OR = 3.7, P = 0.000). CD4-11743 genetic polymorphisms are associated with the susceptibility and severity of RA, and CD4-10845 genetic polymorphisms are associated with the severity of RA. PMID- 21626028 TI - Changes and significance of IL-25 in chicken collagen II-induced experimental arthritis (CIA). AB - Rheumatoid arthritis (RA) is an autoimmune inflammatory disease. It is a systemic inflammatory disease, characterized by chronic, symmetrical, multi-articular synovial arthritis. IL-25 (IL-17E) is a member of the recently emerged cytokine family (IL-17s), which is expressed in Th2 cells and bone marrow-derived mast cells. Unlike the other members of this family, IL-25 is capable of inducing Th2 associated cytokines (IL-4, IL-5, and IL-13) and also promotes the release of some pro-immune factors (IL-6 and IL-8). IL-25 is also a pleiotropic factor, which constitutes a tissue-specific pathological injury and chronic inflammation. In this study, we used chicken collagen II-induced experimental arthritis (CIA) model in DBA/1 mice to investigate the relationship between IL-25 and other inflammatory factors, revealing the possible mechanism in CIA. Our results showed that the expression level of IL-25 was enhanced in the late stage of CIA, and IL 17 was increased in the early stage of the disease. It is well known that IL-17 has a crucial role in the development of RA pathogenesis, and IL-25 plays a significant role in humoral immune. For reasons given above, we suggested that the IL-25 inhibited IL-17 expression to some extent, while enhancing the production of IL-4. It was confirmed that IL-25 not only regulated the cellular immune, but also involved the humoral immune in rheumatoid arthritis. PMID- 21626029 TI - Tumor immunotherapy using adenovirus vaccines in combination with intratumoral doses of CpG ODN. AB - The combination of viral vaccination with intratumoral (IT) administration of CpG ODNs is yet to be investigated as an immunotherapeutic treatment for solid tumors. Here, we show that such a treatment regime can benefit survival of tumor challenged mice. C57BL/6 mice bearing ovalbumin (OVA)-expressing EG.7 thymoma tumors were therapeutically vaccinated with adenovirus type 5 encoding OVA (Ad5 OVA), and the tumors subsequently injected with the immunostimulatory TLR9 agonist, CpG-B ODN 1826 (CpG), 4, 7, 10, and 13 days later. This therapeutic combination resulted in enhanced mean survival times that were more than 3.5* longer than naive mice, and greater than 40% of mice were cured and capable of resisting subsequent tumor challenge. This suggests that an adaptive immune response was generated. Both Ad5-OVA and Ad5-OVA + CpG IT treatments led to significantly increased levels of H-2 K(b)-OVA-specific CD8+ lymphocytes in the peripheral blood and intratumorally. Lymphocyte depletion studies performed in vivo implicated both NK cells and CD8+ lymphocytes as co-contributors to the therapeutic effect. Analysis of tumor infiltrating lymphocytes (TILs) on day 12 post-tumor challenge revealed that mice treated with Ad5-OVA + CpG IT possessed a significantly reduced percentage of regulatory T lymphocytes (Tregs) within the CD4+ lymphocyte population, compared with TILs isolated from mice treated with Ad5-OVA only. In addition, the proportion of CD8+ TILs that were OVA-specific was reproducibly higher in the mice treated with Ad5-OVA + CpG IT compared with other treatment groups. These findings highlight the therapeutic potential of combining intratumoral CpG and vaccination with virus encoding tumor antigen. PMID- 21626030 TI - MAGE-A1, MAGE-A3, and NY-ESO-1 can be upregulated on neuroblastoma cells to facilitate cytotoxic T lymphocyte-mediated tumor cell killing. AB - Approximately half of patients with stage IV neuroblastoma are expected to relapse despite current therapy, and when this occurs, there is little likelihood of achieving a cure. Very few clinical trials have been conducted to determine whether cellular immune responses could be harnessed to fight this tumor, largely because potential tumor antigens for cytotoxic T lymphocytes (CTL) are limited. MAGE-A1, MAGE-A3, and NY-ESO-1 are cancer-testis (CT) antigens expressed on a number of malignant solid tumors, including neuroblastoma, but many tumor cell lines down-regulate the expression of CT antigens as well as major histocompatibility (MHC) antigens, precluding recognition by antigen-specific T cells. If expression of cancer antigens on neuroblastoma could be enhanced pharmacologically, CT antigen-specific immunotherapy could be considered for this tumor. We have demonstrated that the expression of MAGE-A1, MAGE-A3, and NY-ESO-1 can be upregulated on neuroblastoma cells following exposure to pharmacologic levels of the demethylating agent 5-aza-2'-deoxycytidine (decitabine, DAC). Expression of NY-ESO-1, MAGE-A1, or MAGE-A3 was induced in 10/10 neuroblastoma cell lines after 5 days of exposure to DAC. Culture of neuroblastoma cell lines with IFN-gamma was also associated with an increased expression of either MHC Class I or II by cytofluorometry, as reported by other groups. MAGE-A1, MAGE-A3, and NY-ESO-1-specific CTL were cultured from volunteer donors by stimulating peripheral blood mononuclear cells with dendritic cells pulsed with overlapping peptide mixes derived from full-length proteins, and these CTL preferentially lysed HLA partially matched, DAC-treated neuroblastoma and glioblastoma cell lines. These studies show that demethylating chemotherapy can be combined with IFN-gamma to increase the expression of CT antigens and MHC molecules on neuroblastoma cells, and pre-treatment with these agents makes tumor cell lines more susceptible to CTL-mediated killing. These data provide a basis to consider the use of demethylating chemotherapy in neuroblastoma patients, in conjunction with immune therapies that facilitate the expansion of CT antigen-specific CTL. PMID- 21626031 TI - CTL recognition of a novel HLA-A*0201-binding peptide derived from glioblastoma multiforme tumor cells. AB - Genetic instability of tumor cells can result in translation of proteins that are out of frame, resulting in expression of neopeptides. These neopeptides are not self-proteins and therefore should be immunogenic. By eluting peptides from human glioblastoma multiforme (GBM) tumor cell surfaces and subjecting them to tandem mass spectrometry, we identified a novel peptide (KLWGLTPKVTPS) corresponding to a frameshift in the 3' beta-hydroxysteroid dehydrogenase type 7 (HSD3B7) gene. HLA-binding algorithms predicted that a 9-amino acid sequence embedded in this peptide would bind to HLA-A*0201. We confirmed this prediction using an HLA A*0201 refolding assay followed by live cell relative affinity assays, but also showed that the 12-mer binds to HLA-A*0201. Based on the 9-mer sequence, optimized peptide ligands (OPL) were designed and tested for their affinities to HLA-A*0201 and their abilities to elicit anti-peptide and CTL capable of killing GBM in vitro. Wild-type peptides as well as OPL induced anti-peptide CTL as measured by IFN-gamma ELISPOTS. These CTL also killed GBM tumor cells in chromium 51 release assays. This study reports a new CTL target in GBM and further substantiates the concept that rational design and testing of multiple peptides for the same T-cell epitope elicits a broader response among different individuals than single peptide immunization. PMID- 21626032 TI - Camouflage and sabotage: tumor escape from the immune system. AB - The field of tumor immunology has made great progress in understanding tumor immune interactions. As a consequence a number of immuno-therapeutic approaches have been successfully introduced into the clinic and a large number of promising therapeutic strategies are investigated in ongoing clinical trials. Evaluation of anti-tumor immunity in such trials as well as in animal models has shown that tumor escape from immune recognition and tumor-mediated suppression of anti-tumor immunity can pose a significant obstacle to successful cancer therapy. Here, we review mechanisms of tumor immune escape and immune-subversion with a focus on the research interests in our laboratory: loss of MHC class I on tumor cells, increased oxidative stress, recruitment of myeloid-derived suppressor cells, and regulatory T cells. PMID- 21626033 TI - Targeting the extrinsic apoptosis signaling pathway for cancer therapy. AB - The extrinsic apoptosis pathway is triggered by the binding of death ligands of the tumor necrosis factor (TNF) family to their appropriate death receptors (DRs) on the cell surface. One TNF family member, TNF-related apoptosis-inducing ligand (TRAIL or Apo2L), seems to preferentially cause apoptosis of transformed cells and can be systemically administered in the absence of severe toxicity. Therefore, there has been enthusiasm for the use of TRAIL or agonist antibodies to the TRAIL DR4 and DR5 in cancer therapy. Nonetheless, many cancer cells are very resistant to TRAIL apoptosis in vitro. Therefore, there is much interest in identifying compounds that can be combined with TRAIL to amplify its apoptotic effects. In this review, I will provide a brief overview of apoptosis signaling by TRAIL and discuss apoptosis-sensitizing agents, focusing mainly on the proteasome inhibitor bortezomib (VELCADE) and some novel sensitizers that we have recently identified. Alternative ways to administer TRAIL or DR agonist antibodies as therapeutic agents will also be described. Finally, I will discuss some of the gaps in our understanding of TRAIL apoptosis signaling and suggest some research directions that may provide additional information for optimizing the targeting of the extrinsic apoptosis pathway for future cancer therapy. PMID- 21626034 TI - The neuropathological basis of clinical progression in multiple sclerosis. AB - Multiple sclerosis is the major inflammatory condition affecting the central nervous system (CNS) and is characterised by disseminated focal immune-mediated demyelination. Demyelination is accompanied by variable axonal damage and loss and reactive gliosis. It is this pathology that is thought to be responsible for the clinical relapses that often respond well to immunomodulatory therapy. However, the later secondary progressive stage of MS remains largely refractory to treatment and it is widely suggested that accumulating axon loss is responsible for clinical progression. Although initially thought to be a white matter (WM) disease, it is increasingly apparent that extensive pathology is also seen in the grey matter (GM) throughout the CNS. GM pathology is characterised by demyelination in the relative absence of an immune cell infiltrate. Neuronal loss is also seen both in the GM lesions and in unaffected areas of the GM. The slow progressive nature of this later stage combined with the presence of extensive grey matter pathology has led to the suggestion that neurodegeneration might play an increasing role with increasing disease duration. However, there is a paucity of studies that have correlated the pathological features with clinical milestones during secondary progressive MS. Here, we review the contributions that the various types of pathology are likely to make to the increasing neurological deficit in MS. PMID- 21626035 TI - Vasculitis-like neuropathy in amyotrophic lateral sclerosis unresponsive to treatment. AB - Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron disease with variable involvement of other systems. A pathogenetic role of immune-mediated mechanisms has been suggested. We retrospectively analyzed sural nerve pathology and the clinical course in 18 patients with ALS. These patients had undergone sural nerve biopsy because of clinical or neurophysiological signs indicating sensory involvement (ALS+). Eleven of the 18 ALS+ patients had inflammatory cell infiltrates (ALS(vasc)) resembling infiltrates seen in patients with vasculitic neuropathy. Data were compared with the 7 patients without vasculitic infiltrates (ALS(nonvasc)) and with those of 16 patients with isolated peripheral nerve vasculitis (NP(vasc)). Biopsy specimens were processed with standard histological stains and with immunohistochemistry for a panel of inflammatory markers, with the hypothesis that the composition of infiltrates should differ between ALS(vasc) and NP(vasc). Immunoreactive cells were quantified in a blinded manner. Unlike patients with NP(vasc), those with ALS(vasc) had only minor neurophysiological abnormalities in the sural nerve and, except for the infiltrates, almost normal nerve morphology on semithin sections. The difference in epineurial T cell count was significant between ALS(vasc) and ALS(nonvasc) (p = 0.031). Surprisingly, the cellular composition of epineurial infiltrates in sural nerve biopsies was indistinguishable between ALS(vasc) and NP(vasc) despite a significant difference in fiber pathology (p < 0.0001). Standard immunosuppressive treatment did not prevent clinical progression of the motor neuron disease in any of the patients with ALS(vasc). ALS(vasc) appears as a neuropathological subtype in ALS+ suggesting immune-mediated disease components but without response to standard immunosuppressive treatment. PMID- 21626036 TI - Familial amyloidotic polyneuropathy (Portuguese type variant I) and female pelvic floor dysfunction: a tribute to Magellan. PMID- 21626037 TI - Vaginal evisceration in a patient with previous sacrocolpopexy. AB - Vaginal evisceration is a rare gynecologic complication with less than 100 cases reported in the literature. We present a case of vaginal evisceration in a woman with multiple previous surgeries including a sacrocolpopexy. Vaginal mesh grafts have to be applied properly during sacrocolpopexy. PMID- 21626038 TI - Imaging in urogynaecology. AB - The assessment of patients with pelvic floor dysfunction necessitates a combination of clinical skills and adjunct investigations, including detailed imaging. This article reviews a variety of static and dynamic imaging modalities available in the field of urogynaecology, with an emphasis on their clinical implication in identifying the structural and functional causes of pelvic floor disorders. A number of different modalities have been used including X-rays, ultrasound and magnetic resonance imaging. Their place and value are discussed with comments on the validity of the various techniques. PMID- 21626039 TI - Is cesarean section a real panacea to prevent pelvic organ disorders? AB - INTRODUCTION AND HYPOTHESIS: The aim of this study is to compare the pelvic floor muscle strength after vaginal delivery vs. after cesarean section. METHODS: Five groups of 50 cases each were designated as follows: nullipara (control group), spontaneous vaginal delivery (SVD), repeat SVD (SVD-R), cesarean section (CS), and repeat CS (CS-R). Perineometric measurements, stress urinary incontinence (SUI) symptoms, pelvic organ prolapse quantification examinations, and Incontinence-Specific Quality of Life Questionnaire (IQOL) were evaluated. RESULTS: In all delivery groups, pelvic muscle strength was significantly lower than the control group (65.10 +/- 15.69, 56.29 +/- 17.39, 54.28 +/- 20.08, 56.82 +/- 17.62, and 57.92 +/- 16.45 (cm H(2) +/- SD) for the control, SVD, SVD-R, CS, and CS-R groups, respectively; p < 0.05). However, no significant difference was found among the delivery groups. There were significant differences in SUI symptoms between the control (2%) and SVD-R (26%) groups and between the SVD (10%) and SVD-R groups (p < 0.001 and p = 0.037, respectively). No statistically significant correlations between IQOL and perineometric measurements were noted (r = 0.097 and p = 0.598). CONCLUSIONS: Pregnancy increases postpartum muscle weakness independent of the mode of delivery. Increased SUI symptoms are associated with vaginal delivery. PMID- 21626040 TI - Control of bipedal posture following localised muscle fatigue of the plantar flexors and finger-flexors. AB - The present experiment investigated the control of bipedal posture following localised muscle fatigue of the plantar-flexors and finger-flexors. Twelve young healthy adults voluntarily participated in this study. They were asked to stand upright as still as possible with their eyes closed in two randomly ordered experimental sessions. Each session consisted of pre- and post-fatigue bipedal static postural control measurements immediately before and after a designated fatiguing protocol for plantar-flexor and finger-flexor muscles. Centre of foot pressure (CoP) displacements were recorded using a force platform. The results showed that the postural effects of localised muscle fatigue differed between the muscles targeted by the fatiguing procedures. Indeed, localised muscle fatigue of the plantar-flexors yielded increased CoP displacements, whereas localised muscle fatigue of the finger-flexors had no significant effect on the CoP displacements. In other words, fatigue localised to muscles which are involved in the performance of the postural task (plantar-flexors) degraded postural control, whereas fatigue localised to muscles which are not involved in the performance of the postural task did not. Taken together, the present findings support the recent conclusions that the effects of localised muscle fatigue on upright postural control is joint- and/or muscle-specific, and suggest that localised muscles fatigue of the plantar-flexors could mainly affect bipedal postural control via sensorimotor rather than cognitive processes. PMID- 21626041 TI - Artificial gravity training reduces bed rest-induced cardiovascular deconditioning. AB - We studied 15 men (8 treatment, 7 control) before and after 21 days of 6o head down tilt to determine whether daily, 1-h exposures to 1.0 G(z) (at the heart) artificial gravity (AG) would prevent bed rest-induced cardiovascular deconditioning. Testing included echocardiographic analysis of cardiac function, plasma volume (PV), aerobic power (VO(2)pk) and cardiovascular and neuroendocrine responses to 80o head-up tilt (HUT). Data collected during HUT were ECG, stroke volume (SV), blood pressure (BP) and blood for catecholamines and vasoactive hormones. Heart rate (HR), cardiac output (CO), total peripheral resistance, and spectral power of BP and HR were calculated. Bed rest decreased PV, supine and HUT SV, and indices of cardiac function in both groups. Although PV was decreased in control and AG after bed rest, AG attenuated the decrease in orthostatic tolerance [pre- to post-bed rest change; control: -11.8 +/- 2.0, AG: -6.0 +/- 2.8 min (p = 0.012)] and VO(2)pk [pre- to post-bed rest change; control: -0.39 +/- 0.11, AG: -0.17 +/- 0.06 L/min (p = 0.041)]. AG prevented increases in pre-tilt levels of plasma renin activity [pre- to post-bed rest change; control: 1.53 +/- 0.23, AG: -0.07 +/- 0.34 ng/mL/h (p = 0.001)] and angiotensin II [pre- to post bed rest change; control: 3.00 +/- 1.04, AG: -0.63 +/- 0.81 pg/mL (p = 0.009)] and increased HUT aldosterone [post-bed rest; control: 107 +/- 30 pg/mL, AG: 229 +/- 68 pg/mL (p = 0.045)] and norepinephrine [post-bed rest; control: 453 +/- 107, AG: 732 +/- 131 pg/mL (p = 0.003)]. We conclude that AG can mitigate some aspects of bed rest-induced cardiovascular deconditioning, including orthostatic intolerance and aerobic power. Mechanisms of improvement were not cardiac mediated, but likely through improved sympathetic responsiveness to orthostatic stress. PMID- 21626042 TI - Major determinants for the uncovered stent struts on optical coherence tomography after drug-eluting stent implantation. AB - There have been little data regarding major determinants for the uncovered stent struts after drug-eluting stent (DES) implantation on optical coherence tomography (OCT). We investigated the major determinants of incomplete neointimal coverage of DES struts on OCT after implantation in a large cohort of patients. A total of 261 patients with 279 lesions who were treated with various DESs were selected from the OCT registry database. The lesions were divided into two groups based on the ratio of uncovered struts to total struts in all OCT cross-sections; an uncovered group (highest quartile with % uncovered struts >=5.4%, n = 70), and covered group (the remaining lower quartiles with % uncovered struts <5.4%, n = 209). The uncovered group was more likely to have complex lesions, smaller reference vessel and stent diameter, and longer stent, more use of sirolimus eluting stents, and less use of zotarolimus-eluting stents compared with the covered group. Of these variables, the most significant determinant of uncovered stent struts was DES type (odds ratio [OR] = 2.75, 95% confidence interval [CI] = 1.94-3.89, P < 0.001). The use of sirolimus-eluting stents (OR = 2.44, 95% CI, 1.15-5.47, P = 0.023) and zotarolimus-eluting stents (OR = 0.02, 95% CI = 0.01 0.25, P = 0.002) were the only significant risk and protective factors for uncovered stent struts, respectively. This study demonstrated that DES type might be associated with the most important determinants of uncovered struts compared to any other clinical or angiographic factor. PMID- 21626043 TI - Reproducibility of coronary plaque detection and characterization using low radiation dose coronary computed tomographic angiography in patients with intermediate likelihood of coronary artery disease (ReSCAN study). AB - The purpose of this study is to evaluate the interscan, interobserver and intraobserver agreement for coronary plaque detection, and characterization using low radiation dose high-pitch spiral acquisition coronary CT angiography (CTA). Two experienced observers independently evaluated coronary CTA datasets from 50 consecutive patients undergoing two 128-slice dual source CT scans within 12 days. Mean (+/-SD) estimated radiation exposure was 1.5 +/- 0.2 mSv per scan. Observers recorded the presence and characterization of coronary plaques as non calcified or calcified. A "segment involvement score" (SIS) was computed by summing the numbers of segments with any coronary plaque per patient. Reproducibility was assessed using kappa (kappa) statistics, paired t test and Bland-Altman analyses. Interscan, interobserver, and intraobserver agreement (kappa-values) for detection of any or calcified plaques were 83-94% (kappa values 0.57-0.85), and 67-84% (0.31-0.67) for non-calcified plaques on a patient level. No significant difference was observed in mean interscan or interobserver SIS. Mean (95% CI) intraobserver SIS difference was -0.88 (-1.25; -0.51), P < 0.001, with limits of agreement from -4.7 to 2.9. Low radiation dose high-pitch coronary CTA permits detection of any or calcified plaques with high interscan, interobserver, intraobserver agreement. However, variability for the detection of non-calcified plaque is substantial. PMID- 21626044 TI - Image orientation for three-dimensional echocardiography of congenital heart disease. AB - To date there has been little discussion about image orientation for three dimensional (3D) echocardiography when applied to congenital heart lesions. Anatomic relations cannot be assumed in congenital heart disease and image cropping during post processing may by necessity remove external or even internal anatomic references. We present an approach to consistent anatomic orientation which is both intuitive and consistent with regard to superior-inferior, anterior posterior and left-right axes. Such anatomic orientation is also concordant with other common 3D imaging modes such as cardiac magnetic resonance imaging and computed tomography. Views derived from standard cross sectional echocardiography have such universal familiarity that analogous 3D projections of these views may be retained but novel hitherto unavailable views such as en face views of the cardiac septums or atrioventricular valves may be projected using anatomic orientation. PMID- 21626045 TI - Increased mortality in patients with conflicting diastolic parameters. AB - The aim of this study was to describe the clinical characteristics and mortality of patients with conflicting diastolic function during follow-up. Up to 30% of patients have conflicting diastolic function by echo Doppler and therefore cannot be classified into a distinct diastolic dysfunction category of stage 1, 2 or 3. Using our established echocardiography data base, we studied a cohort of 250 subjects with conflicting diastolic function. Each individual was compared to two controls with normal diastolic and systolic function. The pre-specified goal of the analysis was a 6-year follow-up. Patients with conflicting diastolic function were more likely to have diabetes, hypertension, and established coronary artery disease. The Cox proportional hazards model determined that the risk of death was significantly higher for conflicting patients compared to patients with normal diastolic parameters (HR: 1.83; 95% CI: 1.32-2.53), P < 0.001. After adjustment for covariates, the risk of death remained elevated for the conflicting group (HR: 1.56; 95% CI: 1.11-2.18), P = 0.009. Conflicting diastolic dysfunction is associated with an increased risk of death compared to individuals with normal function. In conclusion, this emphasizes the need to attain a more precise characterization and categorization for patients with diastolic dysfunction. PMID- 21626046 TI - Use of the Distress Thermometer to discern clinically relevant quality of life differences in women with breast cancer. AB - PURPOSE: The purpose of this study was to explore the ability of Distress Thermometer (DT) scores to discern important differences in quality of life scores among women with breast cancer. METHODS: The National Comprehensive Cancer Network's DT, the Functional Assessment of Cancer Therapy-Breast (FACT-B), and a demographic questionnaire were completed by 111 women recently diagnosed with breast cancer. RESULTS: Patients considered moderately to severely distressed (score >= 4 on DT) scored significantly lower on FACT-B QOL scales and subscales when compared to those in the group scoring 3 or below. For those scales for which minimally important differences (MIDs) have been established, differences between the two groups were 2-3 and a half times the established MID. CONCLUSIONS: Moderately to severely distressed patients have significantly lower QOL than those with expected or mild distress. The DT provides a quick and easy screening tool to alert the healthcare team to clinically relevant alterations in patients' QOL. PMID- 21626047 TI - Anaphylactic shock and lethal anaphylaxis caused by compound amino acid solution, a nutritional treatment widely used in China. AB - Compound amino acid solution (CAAS) is a large class of solution of amino acids' mixture and was widely used in China. Its extensive nutritional treatment was accompanied by a substantial incidence of adverse reactions, especially life threatening anaphylaxis. However, the adverse reactions were reported in isolated case reports only, and the reasons behind this needed further investigation. The Chinese language papers were searched from China National Knowledge Infrastructure and Wanfang database published in China from 1985 to 2010. The search terms "anaphylactic", "anaphylaxis", "allergic", "allergy", "shock", and "adverse reaction" combined with the term "amino acid" were used. Totally 71 episodes of anaphylactic shock and seven deaths in 38 articles were analyzed. Chest distress and cool extremities were the most common clinical manifestations. Almost all patients suffered from significant hypotension. The vast majority of patients were not found to be allergic to certain substances. CAAS was inappropriately administrated in more than one-third of patients. The life threatening anaphylaxis was prominently prevalent in pregnant women, the elderly and patients with hypersensitivity such as asthma, and patients without medicinal indication. Innovation of processing technique and establishment of more strict supervision system are an urgent need for CAAS to control its production quality and thus improve its safety in China. PMID- 21626048 TI - Pulmonary fibrosis in youth treated with radioiodine for juvenile thyroid cancer and lung metastases after Chernobyl. AB - PURPOSE: The objective of this project was to systematically determine the prevalence and consequences of pulmonary fibrosis in youth with thyroid carcinoma and lung metastases from Belarus who were treated with radioiodine ((131)I). METHODS: A total of 69 patients treated for juvenile thyroid carcinoma and lung metastasis with (131)I were assessed. A group of 29 patients without lung metastases and prior (131)I treatment served as controls. The assessments included a CT scan of the lungs, extensive pulmonary function testing and an incremental cycle test to volitional fatigue with measurements of oxygen uptake (VO(2)), oxygen saturation and alveolar-arterial difference in oxygen partial pressure (DeltaaaO(2)). RESULTS: Five patients with lung metastases showed advanced pulmonary fibrosis on CT scans and also had poorer lung functions compared with the 62 patients with none or minor signs of fibrosis and the 29 controls. Furthermore, these five patients showed lower peak VO(2), lower oxygen saturation at peak exercise and higher exercise DeltaaaO(2). They were younger at the time of cancer diagnosis and had received chemotherapy more frequently than youth with pulmonary metastases who did not develop fibrosis. One of the five patients subsequently died from pulmonary fibrosis. CONCLUSION: Following the Chernobyl catastrophe, about 7% of children treated with radioiodine for thyroid carcinoma and lung metastases displayed pulmonary fibrosis which was associated with functional impairments. Based on the characteristics of affected individuals, the number of radioiodine courses may have to be limited, especially in young children, and chemotherapy should be avoided. PMID- 21626049 TI - A randomized phase II trial of two different 4-drug combinations in advanced pancreatic adenocarcinoma: cisplatin, capecitabine, gemcitabine plus either epirubicin or docetaxel (PEXG or PDXG regimen). AB - PURPOSE: PEFG regimen (P:cisplatin, E:epirubicin, F:5-fluorouracil, G:gemcitabine) significantly prolonged progression-free (PFS) and overall survival (OS) of patients with advanced pancreatic adenocarcinoma (PA) with respect to standard gemcitabine. The current trial was aimed at assessing whether the replacement of E with docetaxel (D) may improve 6 months PFS (PFS6). METHODS: Chemo-naive patients with stage III or metastatic PA received P (30 mg/m(2) day 1 and 15), G (800 mg/m(2) day 1 and 15), and capecitabine (1,250 mg/m(2)/day days 1 28, without a break) and were randomized to receive either D at 25-30 mg/m(2) day 1 and 15 (arm A: PDXG regimen) or E at 30 mg/m(2) day 1 and 15 (arm B: PEXG regimen). Cycles were repeated every 28 days for a maximum of 6 months. The Fleming design was used to calculate the sample size on the probability of being PFS6. Assuming P0 = 40% and P1 = 60%, alpha = 0.05 and beta = 0.10; the study was to enroll 52 patients per arm. RESULTS: Between July 2005 and September 2008, 105 patients were enrolled, stratified by stage and randomized. Patients' characteristics were (A/B) the following: median age 61/59, PS >70 92/88%, metastatic disease 66/65%. PFS6 was 58%, and median OS was 11 months in both arms. A partial response was observed in 60/37% of patients. Main per cycle G3-4 toxicity was the following: neutropenia 4/13%, thrombocytopenia 2/4%, anemia 4/4%, and fatigue 6/3%. CONCLUSIONS: The inclusion of D instead of E yielded more objective response and less G3-4 neutropenia but did not improve PFS and OS. The present trial confirms the relevant impact on outcome of advanced PA of 4-drug regimens. PMID- 21626050 TI - Docetaxel pharmacokinetics and its correlation with two in vivo probes for cytochrome P450 enzymes: the C(14)-erythromycin breath test and the antipyrine clearance test. AB - BACKGROUND: Docetaxel has marked inter-patient PK variability, and metabolic phenotypic probes may enable individualised dosing. This is the first report directly comparing the erythromycin breath test (EBT) (a CYP3A4 probe) with the antipyrine clearance test (ACT), (a general CYP-P450/predominant CYP3A4 probe) for the correlation with docetaxel PK and toxicity. METHODS: Patients pretherapy underwent: (A) EBT: IV C(14)[N-methyl]-erythromycin was administered and breath samples analysed for (14)CO(2), derived parameters included (1) (14)CO(2) flux at 10-min (CO(2)f(10)), (2) 20-min (CO(2)f(20)), (3) terminal rate constant k(CO2) and (4) AUC(CO2,(0-infinity)) and AUC(CO2,(0-60).) (B) ACL test: patients were given oral antipyrine 10 mg/kg, blood samples were taken for PK, and the clearance (CL(Ant)) was derived. Docetaxel was then given at 75 mg/m(2)/3-weekly or 35 mg/m(2)/weekly. Samples taken for docetaxel PK in first course on day 1 and PK parameters included clearance (CL(Doc)). RESULTS: Twenty patients accrued, docetaxel: 3-weekly/weekly = 13:7. EBT parameters (N = 19) (mean, [CV%]): CO(2)f(10) (%/min) 0.051 (106), CO(2)f(20) 0.052 (82), k(CO2) (min(-1)) 0.007 (22), AUC(CO2,(0-infinity)) 7.9 (85), AUC(CO2,(0-60)) 2.64 (81). CL(Ant) (N = 19) (ml/min); 35.8 (37). Docetaxel PK parameters (N = 19): CL(Doc) (l/h) = 57.2 (36), t(Doc1/2) (h) = 12.7 (33). No correlations were observed between the docetaxel PK and EBT parameters. For docetaxel weekly patients, a significant linear relationship was observed between CL(Doc) and CL(Ant) (P = 0.007, R (2) = 79.47%). CONCLUSIONS: The utility of EBT for the prediction of docetaxel PK was not confirmed in this study. The antipyrine clearance test may be superior in this regard for docetaxel, but regimen dependent and hence warrants further evaluation. PMID- 21626051 TI - A drug interaction study evaluating the pharmacokinetics and toxicity of sorafenib in combination with capecitabine. AB - PURPOSE: To address tolerability and a possible pharmacologic interaction of capecitabine with sorafenib. METHODS: Patients with advanced solid tumors (ECOG PS 0-1) were included. Cohort A received capecitabine 750 mg/m(2) BID and Cohort B received capecitabine 1,000 mg/m(2) BID, both for 14 days of a 21-day cycle. Steady-state PK was obtained for capecitabine alone, sorafenib alone, and in combination. Cohort C explored an alternate schedule of 7-day on/7-day off flat dose capecitabine 1,000 mg BID with continuous dosing of sorafenib 400 mg BID. RESULTS: A total of 32 patients were enrolled between February 08 and April 09. Hand-foot skin reaction (HFSR) was the primary toxicity with 16 (50%) of the 32 patients experiencing grade 3 events (75% occurring during cycles 1-2). Grade 3 HFSR defined the maximum tolerated dose (MTD) of Cohort C at 1,000 mg BID flat dose of capecitabine. Other grade 3/4 toxicities were rare (diarrhea 6%, mucositis 3%, and fatigue 3%). Capecitabine did not change the C (max) or AUC((0 12)) of sorafenib. Co-administration of sorafenib with capecitabine 750 mg/m(2) (n = 6 patients) increased capecitabine AUC((0-12)) 15% and produced no change in the 5FU AUC((0-12)). At the capecitabine 1,000 mg/m(2) dose level (n = 12 pts), there was a 16% increase in capecitabine AUC((0-12)) and an 8% increase in 5FU AUC((0-12)). However, these trends were not statistically significant. CONCLUSIONS: Co-administration of sorafenib resulted in a mild increase in capecitabine AUC, although not statistically significant. Capecitabine did not affect the exposure of sorafenib. The rate of grade 3 HFSR is concerning and limits the feasibility of prolonged dosing of sorafenib with capecitabine 1,000 mg/m(2) on the 21-day schedule. PMID- 21626052 TI - "Guardarse las cosas adentro" (keeping things inside): Latina violence survivors' perceptions of depression. AB - BACKGROUND: Latinos are less likely than non-Hispanic whites to be adequately treated for depression. Intimate partner violence (IPV) is strongly associated with depression. Less is known about how Latina IPV survivors understand depression. OBJECTIVE: To understand Latina women's beliefs, attitudes, and recommendations regarding depression and depression care, with a special focus on the impact of gender, ethnicity, violence, and social stressors. DESIGN: Focus group study. PARTICIPANTS: Spanish-speaking Latina women with a lifetime history of IPV and moderate to severe depressive symptoms. APPROACH: We used a community based participatory research (CBPR) approach to conduct a thematic analysis using an inductive approach. RESULTS: Thirty-one women participated in five focus groups. Women felt depression is caused by "keeping things inside". They also felt that keeping things inside could lead to physical illness or an inability to function. Their inability to talk was fueled by issues such as stigma, fear, isolation, cultural norms, or simply "not having the words". They felt that the key to treating depression was finding a way to talk about the things that they had kept inside. They greatly valued information about depression and appreciated learning from providers that their physical symptoms were caused by depression. They wanted confidential depression care programs that not only helped them deal with their depression, but also addressed the violence in their lives, gave them practical skills, and attended to practical issues such as childcare. They had negative attitudes toward antidepressants, primarily due to experiences with side effects. Negative experiences with the health care system were primarily attributed to lack of good healthcare insurance. CONCLUSIONS: The concept of "keeping things inside" was key to participants' understanding of the cause of depression and other health problems. Clinicians and depression care programs can potentially use such information to provide culturally-appropriate depression care to Latina women. PMID- 21626054 TI - The Autism-Spectrum Quotient--Italian version: a cross-cultural confirmation of the broader autism phenotype. AB - The Autism Spectrum Quotient (AQ) has been used to define the 'broader' (BAP), 'medium' (MAP) and 'narrow' autism phenotypes (NAP). We used a new Italian version of the AQ to test if difference on AQ scores and the distribution of BAP, MAP and NAP in autism parents (n = 245) versus control parents (n = 300) were replicated in a Sicilian sample. Parents of children with autism spectrum conditions scored higher than the control parents on total AQ, social skills and communication subscales, and exhibited higher rates of BAP, MAP and NAP. We conclude that the Italian AQ is a cross-culturally reliable measure of these different phenotypes, and can be used to identify a phenotypic gradient of severity of autistic traits in families. To understand the molecular basis of these phenotypes will require its use in genetic association studies. PMID- 21626055 TI - Analysis of an Egyptian study on the socioeconomic distribution of depressive symptoms among undergraduates. AB - BACKGROUND: Worldwide population surveys show that prevalence of mental disorders in developing countries was as common as in industrialized countries. College life is an important stage in every person's life; unfortunately, it may be the time to experience depression for the first time. Latterly, there has been a steady increase in the number of university students diagnosed and treated as depressed patients. This paper presents the results of a study carried out on a representative sample of undergraduate students of an Egyptian University, which aimed to study the relationship between social factors and the occurrence of depression. It was assumed that the poorer the socio-economic background the higher the rates of depression. METHOD: Data were collected in 2004 from students at Assiut University using the Zagazig Depression scale (ZDS) and an Egyptian socio-economic status measure. This analysis, based on a refined version of the ZDS used STATA (v. 10.1.). Descriptive statistics, univariate and multivariate regression models were used to test for associations with the dimensions of SES and depression. RESULT: Thirty-seven percent of the students had symptoms scored above the threshold for moderate depression. In the fully adjusted model: faculty of study, father's occupation, family income and number of persons per room were associated with depression in this sample. CONCLUSION: The high prevalence of depressive symptoms among Egyptian students found in this study call attention to the need for a student counseling service offering mental health support. This service should particularly discover an approach for low affluence students. PMID- 21626056 TI - Labels used by young people to describe mental disorders: which ones predict effective help-seeking choices? AB - PURPOSE: Mental disorders are common in young people, yet many do not seek help. Being able to label the problem may facilitate effective help-seeking, but it is not clear which labels are best. This study aims to examine which labels commonly used by young people are associated with a preference for recommended sources of help and treatment. METHOD: A national telephone survey was conducted with a randomly selected sample of 2,802 Australian young people aged 12-25 years. Respondents were read out one of three vignettes describing symptoms of a mental disorder, and asked a series of questions regarding labelling of the problem described and related help-seeking preferences and beliefs. Binary logistic regression analyses were used to measure the association between type of label used and help-seeking preferences and beliefs. RESULTS: Use of the accurate label to describe the problem in the vignette predicted a preference for recommended sources of help with greater consistency than any other labels commonly used by young people. Inaccurate mental health labels did predict some preferences for recommended sources of help and treatment, but not to the extent of the accurate label. Lay labels such as "stress", "paranoid" and "shy" predicted less intention to seek any help for the problem described in the vignette. CONCLUSIONS: Labelling a disorder accurately does predict a preference for recommended sources of help and a belief in the helpfulness of recommended treatments. Importantly, it is also apparent that some commonly used lay labels cannot do this and indeed may limit appropriate help-seeking and treatment acceptance. PMID- 21626057 TI - Service user attachments to psychiatric key workers and teams. AB - PURPOSE: The first aim of the study was to evaluate whether self-reported attachment styles of individuals with psychosis are consistent with their self reported attachment in therapeutic relationships with both key workers and mental health teams. The second aim was to evaluate the level of concordance in attachment ratings given by different raters (self-report, key worker informant report and team informant-report). METHODS: Three self-report versions of the Psychosis Attachment Measure (PAM; attachment in general relationships, attachment towards key worker and attachment in relation to the mental health team) were administered to 24 individuals with a diagnosis of psychosis in psychiatric rehabilitation settings. Key worker and 'team' informant versions of the PAM were also completed. RESULTS: There were strong, significant correlations among the three self-reported attachment measures. There was less consistent evidence of correlations between key worker ratings of attachment and self-report attachment ratings. The majority of the correlations between team ratings of attachment and self-report attachment were small and non-significant. CONCLUSIONS: Strong correlations among the self-reported PAM scales suggest that self-reported attachment in specific therapeutic relationships is consistent with self-reported attachment in general relationships. The self-ratings were not consistently correlated with informant-ratings and team informant ratings were particularly poorly correlated with self-ratings. This suggests that it is vital that teams consult service users themselves when making decisions about their care. PMID- 21626058 TI - Generating small-area prevalence of psychological distress and alcohol consumption: validation of a spatial microsimulation method. AB - PURPOSE: Public mental health surveillance data are rarely available at a fine geographic scale. This study applies a spatial microsimulation procedure to generate small-area (lower super outputs areas [LSOA]) estimates of psychological distress and alcohol consumption. The validity of LSOA estimates and their associations with proximal and broader socioeconomic conditions are examined. METHODS: A deterministic reweighting methodology assigns prevalence estimates for psychological distress and heavy alcohol consumption through a process of matching individuals from a large, population-representative dataset (Health Survey for England) to known LSOA populations (from the 2001 population Census). 'goodness-of-fit' of LSOA estimates is assessed by their comparison to observed prevalence of these health indicators at higher levels of aggregation (local authority districts [LAD]). Population prevalence estimates are correlated to the mental health needs index (MINI) and other health indicators; ordered logistic regression is applied to investigate their associations with proximal and broader socioeconomic conditions. RESULTS: Performance of microsimulation models is high with no more than 10% errors in at least 90% of LAD for psychological distress and moderate and heavy alcohol consumption. The MINI is strongly correlated with psychological distress (r = 0.910; p value < 0.001) and moderately with heavy drinking (r = 0.389; p value < 0.001). Psychological distress and heavy alcohol consumption are differently associated with socioeconomic and rurality indicators at the LSOA level. Associations further vary at the LAD level and regional variations are apparent. CONCLUSION: Spatial microsimulation may be an appropriate methodological approach for replicating social and demographic health patterns at the local level. PMID- 21626059 TI - Design of dry nanosuspension with highly spontaneous dispersible characteristics to develop solubilized formulation for poorly water-soluble drugs. AB - PURPOSE: The powderization of the aqueous nanosuspension of a poorly water soluble drug, which was prepared by wet-milling technique developed by authors, was investigated to apply to the development of solid dosage forms. METHODS: Drug particles were suspended and milled in the aqueous medium using the oscillating beads-milling apparatus. The recovered nanosuspension was spray-dried 1) with no additive or 2) with co-dissolving mannitol as a nanoparticle carrier. As a control, the solution of the drug and additives with the same formulation as nanosuspension was also spray-dried. RESULTS: SEM observation and X-ray powder diffraction analysis revealed that the dried products from suspension formed a spherical particle with single-micron diameter, which was composed of thousands of nano-sized crystalline drug fragment. It was also found that the dried products from suspension could be spontaneously redispersed in water, transforming into nanosuspension with the original size distribution. Such dried powder with high dispersibility was named "dry nanosuspension." The dry nanosuspension had immediate release behaviors in gastrointestinal buffered media, whereas the dried product from solution showed the poor dispersion and dissolution properties even if same content of additives were loaded. CONCLUSIONS: The present technique with combination of wet nano-milling and spray drying processes would be a novel approach to develop the pharmaceutical products with poorly water-soluble and oral-absorbable drugs. PMID- 21626060 TI - Establishing a link between amino acid sequences and self-associating and viscoelastic behavior of two closely related monoclonal antibodies. AB - PURPOSE: To investigate the underlying cause for the observed differences in self associating and viscoelastic behavior between two monoclonal antibodies, MAb1, and MAb2. METHODS: Several mutants were designed by swapping charged residues in MAb1 with those present in MAb2 at their respective positions and vice versa. Rheological analysis was done at low and high shear rates. Dynamic light scattering quantified intermolecular interactions in dilute solutions; sedimentation equilibrium analysis determined the corrected weight average molecular weight (M (wc)) to assess the self-associating behavior in high concentration. The molecular charge was estimated from electrophoretic mobility measurements. RESULTS: Replacing the charged residues in the CDR of MAb1 resulted in a lower M (wc) and solution viscosity. The corresponding changes in either just the variable light (VL) or variable heavy (VH) chain showed only a partial decrease in viscosity, whereas changes in both VL and VH chains resulted in a dramatic reduction in viscosity. The converse case where the VL and VH chains of MAb2 were made to look like MAb1 did not self-associate or show increased viscosity. CONCLUSIONS: Exposed charged residues in the CDR of MAb1 are critical in determining the self-associating and highly viscous behavior observed at high concentrations. PMID- 21626061 TI - PLGA microparticles encapsulating prostaglandin E1-hydroxypropyl-beta cyclodextrin (PGE1-HPbetaCD) complex for the treatment of pulmonary arterial hypertension (PAH). AB - PURPOSE: To test the efficacy and viability of poly (lactic-co-glycolic acid) (PLGA) microspheres encapsulating an inclusion complex of prostaglandin E(1) (PGE(1)) and 2-hydroxypropyl-beta-cyclodextrin (HPbetaCD) for pulmonary delivery of PGE(1) for treatment of pulmonary arterial hypertension (PAH), a disease of pulmonary circulation. METHODS: PLGA-based microparticulate formulations of PGE(1)-HPbetaCD inclusion complex or plain PGE(1) were prepared by a double emulsion solvent evaporation method. HPbetaCD was used as a complexing agent to increase the aqueous solubility of PGE(1), act as a porosigen to produce large porous particles, and promote absorption of PGE(1). Particles were characterized for micromeritic properties, in vivo absorption, metabolic degradation, and acute safety. RESULTS: Incorporation of HPbetaCD in the microparticles resulted in development of large particles with internal pores, which, despite large mean diameters, had aerodynamic diameters in the inhalable range of 1 to 5 MUm. HPbetaCD incorporation also resulted in a significant increase in the amount of drug released in vitro in simulated interstitial lung fluid, showing a desirable burst release profile required for immediate hemodynamic effects. Compared to plain PLGA microparticles, entrapment efficiency was decreased upon complexation with HPbetaCD. In vivo absorption profile indicated prolonged availability of PGE(1) in circulation following pulmonary administration of the optimized microparticulate formulations, with an extended half-life of almost 4 hours. Metabolic degradation and acute toxicity studies suggested that microparticulate formulations were stable under physiological conditions and safe for the lungs and respiratory epithelium. CONCLUSIONS: This study demonstrates the feasibility of PGE(1)-HPbetaCD complex encapsulated in PLGA microparticles as a potential delivery system for controlled release of inhaled PGE(1). PMID- 21626062 TI - Isolation and molecular characterization of canine distemper virus from India. AB - Ocular swabs from canine distemper virus (CDV) suspected live or brain tissue from dead dogs were tested for the presence of CDV nucleoprotein (N) gene using reverse transcriptase polymerase chain reaction (RT-PCR). Partial "N" gene sequencing of the RT-PCR-positive samples and the local vaccine virus revealed that the Ind/Andaman 01/07 virus was highly divergent from the rest of the CDV isolates and from the vaccine strain. Quantitative real-time PCR (qRT-PCR) using SYBR Green I chemistry for CDV haemagglutinin "H" gene quantification showed C(t) values ranging from 29.76-30.67 in the RT-PCR-positive samples. Two of the positive samples, designated Ind/TN 01/07 and Ind/Andaman 01/07 were used for virus isolation in B95a cell line. Characteristic cytopathic changes such as rounding of cells, syncytia formation, and ballooning were seen from the first passage onwards. Specific cytoplasmic fluorescence was seen in infected cells with a commercial reference serum against CDV. To the best of our knowledge, this is the first report of CDV isolation from clinical cases in India. PMID- 21626063 TI - Immunohistochemical and histopathological findings of ovine pulmonary adenocarcinoma (Jaagsiekte) in Egyptian sheep. AB - Ovine pulmonary adenocarcinoma (OPA) is a naturally occurring retrovirus-induced transmissible lung cancer in sheep. Lungs and associated (bronchial and mediastinal) lymph nodes of seven sheep with OPA were examined. Lungs had few multifocal consolidated slightly elevated gray to white masses ranging from 0.5 to 3 cm in diameter. Histopathologically, these masses appeared as well differentiated acinar adenocarcinoma with little evidence of anaplasia. The acini composed of well-differentiated cuboidal to low columnar epithelium with clear or vacuolated cytoplasm and low mitotic index. No metastases were observed in the bronchial and mediastinal lymph nodes of any animal. The presence of Jaagsiekte sheep retrovirus (JSRV) was demonstrated in the lungs by immunohistochemistry. JSRV protein was detected in all tumor epithelial cells, histologically normal alveolar type II cells, and few bronchiolar epithelial cells, alveolar macrophages, lymphocytes, and plasma cells. This study is the first to confirm the presence of natural OPA in Egypt. PMID- 21626064 TI - Prevalence and risk factors for subclinical coccidiosis in broiler chicken farms in Mazandaran province, Iran. AB - Coccidiosis is a disease of almost universal importance in Poultry production. The subclinical form of coccidiosis has most significant economic impact due to impaired growth rate and feed conversion. Some factors such as: age, size of flock, season, etc., may effect on severity of this disease. In this study, the relationships between some major risk factors and prevalence of subclinical coccidiosis were investigated. This study was done in 120 broiler farms in five different cities of Mazandaran province, north of Iran. Five chicks (3-8 weeks of ages) were taken randomly from every 120 farms and post-mortem and parasitological examinations were performed. Five Eimeria spp. were recognized: Eimeria tenella, Eimeria maxima, Eimeria acervulina, Eimeria brunetti, and Eimeria necatrix. The prevalence rate of subclinical coccidiosis among them is 75% (90 farms out of 120). E. acervulina was the most prevalent species (65.5%) followed by E. maxima (17.7%), E. tenella (15.5%), E. brunetti (10%), and E. necatrix (5.5%). According to what the results approve, the occurrence of subclinical coccidiosis is significantly related to the age and size of flock, whereas the other factors such as the season of year, industrial strains, chicken's keeping system, and anticoccidial drugs do not affect this phenomenon remarkably. PMID- 21626065 TI - In vitro predatory activity of nematophagous fungi Duddingtonia flagrans on infective larvae of Oesophagostomum spp. after passing through gastrointestinal tract of pigs. AB - One isolate of predator fungi Duddingtonia flagrans (AC001) was assessed in vitro regarding the capacity of supporting the passage through pigs' gastrointestinal tract without loss of the ability of preying infective larvae Oesophagostomum spp. Fungal isolates survived the passage and were efficient in preying L(3) since the first 8 h of collection (p < 0.01) in relation to the control group (without fungus). Compared with control, there was a significant decrease (p < 0.01) of 59.6% (8 h), 71.7% (12 h), 76.8% (24 h), 81.0% (36 h), 78.0% (48 h), 76.1% (72 h), and 82.7% (96 h) in means of infective larvae Oesophagostomum spp. recovered from treatments with isolate AC001. Linear regression coefficients of L(3) of recovered Oesophagostomum spp. regarding the collections due to time were -0.621 for control, -1.40 for AC001, and -2.64 for NF34. Fungi D. flagrans (AC001) had demonstrated to be promising for use in the biological control of pig parasite Oesophagostomum spp. PMID- 21626066 TI - Serological and molecular detection of Mycobacterium avium subsp. paratuberculosis in cattle of dairy herds in Colombia. AB - The objective of this study is the detection of Mycobacterium avium subsp. paratuberculosis (MAP) by serum enzyme-linked immunosorbent assay (ELISA), fecal polymerase chain reaction (PCR), and fecal culture in Colombian dairy herds. Serum and fecal samples from asymptomatic cows (n = 307) of 14 dairy herds were tested for MAP by an unabsorbed ELISA test (ELISA-A). Serum and fecal samples from positive ELISA-A animals (n = 31) were further tested by an absorbed ELISA test (ELISA-B) and PCR. Fecal samples from animals of herds positive by ELISA-A and PCR (n = 105) were inoculated onto three different culture media. ELISA-A produced positive results in 10% of the serum samples and 71% of the herds. ELISA B and PCR results were positive in two and six serum and fecal samples from positive ELISA-A animals, respectively. Fecal samples were negative for MAP on all culture media. The results of this study confirmed the presence of MAP in local dairy herds and the difficulties of MAP detection in asymptomatic animals by ELISA, PCR, and fecal culture. PMID- 21626067 TI - Are two heuristics better than one? The fluency and distinctiveness heuristics in recognition memory. AB - Four experiments were conducted to test the impact of having multiple heuristics (distinctiveness and fluency) available during a recognition test. Recent work by Gallo, Perlmutter, Moore, and Schacter (Memory & Cognition 36:461-466, 2008) suggested that fluency effects are reduced when the distinctiveness heuristic can be applied to a recognition decision. In Experiment 1, we used a response reversal paradigm (Van Zandt & Maldonado-Molina Journal of Experimental Psychology: Learning, Memory, and Cognition 30:1147-1166, 2004) to demonstrate that participants transitioned from an early response strategy that was largely reliant on fluency to a later strategy in which the influences of fluency and distinctiveness were both observable. Experiments 2a, 2b, and 3 showed no evidence for reduction of the fluency heuristic after picture study when the test required a delayed response (Exp. 2a), confidence ratings (Exp. 2b), or the application of conceptual fluency (Exp. 3). The results are consistent with models of memory that assume that familiarity and recollection influence individual memory decisions Wixted (Psychological Review, 114:152-176, 2007). PMID- 21626068 TI - Traces of times past: representations of temporal intervals in memory. AB - Theories of time perception typically assume that some sort of memory represents time intervals. This memory component is typically underdeveloped in theories of time perception. Following earlier work that suggested that representations of different time intervals contaminate each other (Grondin, 2005; Jazayeri & Shadlen, 2010; Jones & Wearden, 2004), an experiment was conducted in which subjects had to alternate in reproducing two intervals. In two conditions of the experiment, the duration of one of the intervals changed over the experiment, forcing subjects to adjust their representation of that interval, while keeping the other constant. The results show that the adjustment of one interval carried over to the other interval, indicating that subjects were not able to completely separate the two representations. We propose a temporal reference memory that is based on existing memory models (Anderson, 1990). Our model assumes that the representation of an interval is based on a pool of recent experiences. In a series of simulations, we show that our pool model fits the data, while two alternative models that have previously been proposed do not. PMID- 21626069 TI - Primary intraocular lymphoma: a review. AB - Primary intraocular lymphoma (PIOL) is a rare, non-Hodgkin lymphoma considered to be a subtype of primary central nervous system lymphoma. We describe a 65-year old woman who presented to the Hematology/Oncology Clinic at Scripps Clinic, La Jolla, California, who was diagnosed with bilateral PIOL without systemic disease. She enjoyed a 16-month remission but ultimately recurred in the brain. We reviewed the literature and present a discussion of the diagnostic criteria for PIOL and current strategies for treating PIOL in immunocompetent patients. PMID- 21626070 TI - Outcome and prognostic features in anaplastic ganglioglioma: analysis of cases from the SEER database. AB - Anaplastic ganglioglioma (AGG) are rare central nervous system tumours. Patient and treatment factors associated with outcome are poorly defined and limited to small retrospective case series and single case reports. Using the Surveillance, Epidemiology, and End Results (SEER) cancer registry, we investigated potential clinicopathological factors that can affect outcome in patients with anaplastic ganglioglioma. Patients with anaplastic ganglioglioma diagnosed between 1973 and 2007 were identified from the SEER database. Kaplan-Meier survival analysis and Cox models were used to examine the effect of variables on overall survival. The variables analysed included patient age at diagnosis, gender, race, tumour location, uni-focal or multi-focal tumour, surgical resection and the use of adjuvant radiotherapy. Fifty-eight patients were identified, with a median age at diagnosis of 25.5 years. Ninety-three percent of patients underwent surgery and 36% received adjuvant radiotherapy. The median overall survival was 28.5 months. The most common tumour site was the temporal lobe (27%). Univariate and multivariate analysis identified surgery and uni-focal disease as important predictors of overall survival. Adjuvant radiotherapy did not influence overall survival. This study represents the largest analysis of anaplastic ganglioglioma to date. Furthermore it also emphasises the role of national tumour databases for furthering our understanding of rare brain tumours and determining management options. PMID- 21626071 TI - Salvage therapy with single agent bendamustine for recurrent glioblastoma. AB - The treatment of recurrent glioblastoma (GBM) remains challenging notwithstanding the recent approval of bevacizumab for this indication. Bendamustine has a bifunctional mechanism of action including alkylation, penetrates the CNS and does not show cross resistance to other alkylator chemotherapies. In a single institution phase 2 trial, patients with recurrent GBM were treated with bendamustine (100 mg/m(2)/day administered intravenously for two consecutive days every 4 weeks). The primary study endpoint was 6-month progression free survival (PFS-6). An interim analysis for futility was conducted according to a Simon two stage minimax design. Complete blood counts were obtained bimonthly, clinical evaluations and brain imaging every month for the first cycle and bimonthly thereafter. Treatment responses were based upon MacDonald criteria. Sixteen patients were enrolled (nine men; seven women), with a median age of 53 years (range 36-68) and a median Karnofsky performance status of 90 (range 70-100). Nine patients were treated at first relapse and seven at second relapse (five patients were bevacizumab failures). A total of 25 cycles of bendamustine were administered with a median of 1 (range 1-6). Bendamustine-related toxicity was seen in eight patients; lymphopenia in seven (5 grade 3; 2 Grade 4), thrombocytopenia in two (1 Grade 3; 1 Grade 4), and neutropenia in one (1 Grade 3). Fourteen patients have died due to disease progression, two patients are alive and on alternative therapies. Only one patient was progression-free at 6 months, triggering the stopping rule for futility. Bendamustine was reasonably well tolerated but failed to meet the study criteria for activity in adults with recurrent GBM. PMID- 21626072 TI - Leptomeningeal dissemination of anaplastic glioma: prolonged survival in two patients treated with temozolomide. PMID- 21626073 TI - Assessment of MGMT promoter methylation status in pleomorphic xanthoastrocytoma. AB - Although pleomorphic xanthoastrocytoma (PXA) is currently designed as a grade II glioma according to the WHO classification, a significant percentage of the tumors undergo malignant progression. In this study, the MGMT methylation status was examined in 11 PXA patients to determine if a biologic rationale exists to support the use of temozolomide (TMZ) for treatment of aggressive PXA. There were 9 cases of PXA grade II and 2 cases of PXA with anaplastic features. In the MGMT methylation study, only 2 (18.1%) of the 11 tumor samples tested by MS-qLNAPCR were positive for MGMT promoter methylation. In contrast, other cases, including PXAs with anaplastic features, were unmethylated. In addition, a tumor recurrence was found to be unmethylated. Thus, MGMT promoter methylation is not frequent in PXA and our results raise doubts about the benefits of treating indistinctly aggressive PXA with TMZ. PMID- 21626075 TI - A patient with rheumatoid arthritis treated with tocilizumab together with lamivudine prophylaxis after remission of infliximab-reactivated hepatitis B. AB - A 59-year-old woman with rheumatoid arthritis was treated quite successfully with infliximab, but her serum aminotransferase levels were markedly elevated; this was diagnosed as acute exacerbation of hepatitis B and she was treated with lamivudine, and infliximab was discontinued. The rheumatoid arthritis disease activity was uncontrollable after the discontinuation of infliximab, and we therefore initiated tocilizumab treatment (after obtaining the patient's informed consent) together with lamivudine prophylaxis. After tocilizumab administration her rheumatoid arthritis disease activity was significantly attenuated, and the activity has remained low, without re-exacerbation of the hepatitis, for more than 2 years since the initiation of the tocilizumab. PMID- 21626076 TI - Critically ill patients and end-of-life decision-making: the senior medical resident experience. AB - In order to improve the understanding of educational needs among residents caring for the critically ill, narrative accounts of 19 senior physician trainees participating in level of care decision-making were analyzed. In this multicentre qualitative study involving 9 university centers in Canada, in-depth interviews were conducted in either English or French, and the transcripts then underwent a hermeneutic phenomenological analysis. The resident was the central figure in the narrated incident, along with the patients' relatives and other attending physicians. The vast majority of interviews recounted negative experiences that involved delivering bad news to patients' families and managing difficult communications with them and with physician colleagues. Emotional distress and suffering were often part of their decision-making process. Narrating their experiences was viewed as a positive event. Data analysis uncovered 6 general themes that were organized into 2 categories, the first one grouping together themes related to interactions with the patients' families and the second comprising themes related to interactions with physician coworkers. The findings suggest that physician trainees' narratives are a rich source of data regarding what constitutes meaningful training experiences and what they learn from them. Educational approaches that incorporate the telling of stories would allow students to express their feelings, doubts, and opinions about their work experiences and could thus foster personal and emotional learning in critical care. PMID- 21626077 TI - Antihypertensive Treatment of Acute Cerebral Hemorrhage (ATACH) II: design, methods, and rationale. AB - The December 2003 report from the National Institute of Neurological Disorders and Stroke (NINDS) Workshop on priorities for clinical research in intracerebral hemorrhage (ICH) recommended clinical trials for evaluation of blood pressure management in acute ICH as a leading priority. The Special Writing Group of the Stroke Council of the American Heart Association in 1999 and 2007 emphasized the need for clinical trials to ensure evidence-based treatment of acute hypertensive response in ICH. To address important gaps in knowledge, we conducted a pilot study funded by the NINDS, Antihypertensive Treatment of Acute Cerebral Hemorrhage (ATACH) I Trial, during 2004-2008 to determine the appropriate level of systolic blood pressure (SBP) reduction. We now have initiated a multi-center, randomized Phase III trial, the ATACH II Trial, to definitively determine the efficacy of early, intensive, antihypertensive treatment using intravenous (IV) nicardipine initiated within 3 h of onset of ICH and continued for the next 24 h in subjects with spontaneous supratentorial ICH. The primary hypothesis of this large (N = 1,280), streamlined, and focused trial is that SBP reduction to <=140 mm Hg reduces the likelihood of death or disability at 3 months after ICH, defined by modified Rankin scale score of 4-6, by at least 10% absolute compared to standard SBP reduction to <=180 mm Hg. The ATACH II trial is a natural extension of numerous case series, the subsequent ATACH I pilot trial, and a preliminary, randomized, and controlled trial in this patient population funded by the Australian National Health and Medical Research Council. Both trials recently confirmed the safety and tolerability of both the regimen and goals of antihypertensive treatment in acutely hypertensive patients with ICH, as proposed in the present trial. The underlying mechanism for this expected beneficial effect of intensive treatment is presumably mediated through reduction of the rate and magnitude of hematoma expansion observed in approximately 73% of the patients with acute ICH. The Australian trial provided preliminary evidence of attenuation of hematoma expansion with intensive SBP reduction. The ATACH II trial will have important public health implications by providing evidence of, or lack thereof, regarding the efficacy and safety of acute antihypertensive treatment in subjects with ICH. This treatment represents a strategy that can be made widely available without the need for specialized equipment and personnel, and therefore, can make a major impact upon clinical practice for treating patients with ICH. PMID- 21626078 TI - Locoregional therapy-induced tumor necrosis as a predictor of recurrence after liver transplant in patients with hepatocellular carcinoma. AB - BACKGROUND: Patients with hepatocellular carcinoma (HCC) often undergo locoregional therapy before liver transplant either to downstage the tumor or as bridge therapy. Our goal was to assess the risk factors for posttransplant tumor recurrence, specifically the extent of necrosis induced by locoregional therapy. METHODS: We conducted a hospital-based retrospective analysis of 100 patients with HCC who received a liver transplant, 86 of whom had received pretransplant locoregional therapy. We evaluated various patient- and tumor-related parameters to determine the risk factors for recurrence. Furthermore, we grouped patients by the degree of tumor necrosis after locoregional therapy and identified the factors that were associated with a favorable tumor response. RESULTS: Initial tumor extent beyond the University of San Francisco (UCSF) criteria, microvascular invasion, and attainment of less than 90% tumor necrosis after locoregional therapy were independent risk factors for tumor recurrence. In addition, there was a significant correlation between the tumor necrosis percentage and disease-specific survival rate. Among patients whose tumors initially exceeded the UCSF criteria, those with extensive locoregional therapy induced tumor necrosis had lower recurrence rates. All recurrences after transplant occurred within 3 years, and recurrence rates in patients with extensive tumor necrosis at 1, 2, and 3 years were 3%, 6%, and 10%, respectively. Female gender and a solitary tumor were independently associated with extensive tumor necrosis. CONCLUSIONS: In HCC patients who are transplant candidates and undergo pretransplant locoregional therapy, the degree of induced tumor necrosis affects both tumor recurrence and survival rate. PMID- 21626079 TI - Sentinel lymph node pressure in breast cancer. AB - BACKGROUND: Leakiness of angiogenic tumor vessels results in elevated pressure in primary breast cancers and increased lymphatic flow to sentinel lymph node(s) (SLNs). We hypothesized that a similar pathophysiology in metastatic axillary SLNs would result in increased intranodal pressure (INP). METHODS: SLNs were "hot" and "blue" after intramammary injection of dilute methylene blue and filtered Tc99 sulfur colloid. Intraoperative pressure was measured in SLNs by a noncoring needle and recording device in 114 breast cancer patients. Excised axillary SLNs were examined by standard pathological techniques and metastases measured, recorded, and compared with INP measurements for SLN #1 and sometimes #2. RESULTS: INP in 131 SLNs with no tumor (SLN #1, n = 93; SLN #2, n = 38) was 9.1 +/- 6.2 (SD; range -2, 35) mmHg and 21.4 +/- 15.4 mmHg (range 0-50) in 35 tumor-containing SLNs (SLN #1, n = 29; SLN #2, n = 6) (P = 0.0066). Elevated INPs significantly correlated with SLN tumor metastasis sizes (P = 0.0038; r = 0.4904). In two patients, tumor-laden SLNs with high INP were not blue or "hot" while a blue lymphatic bypassed these nodes and was traced to the next echelon tumor-free blue and "hot" nodes with low INP. CONCLUSIONS: Breast cancer metastasis in axillary SLNs was associated with significantly higher INP than in tumor-free lymph nodes. When "true" SLNs were replaced by tumor, and the INP levels were very high, lymph flow direction changed; lymphophilic particles (blue dye and radiocolloid) were redirected to the next echelon of nodes, where the pressures were much lower. Mechanical factors may increase the likelihood of metastasis to neighboring lymph nodes with lower INP. PMID- 21626080 TI - Simultaneous medullary and papillary thyroid cancer: a novel entity? AB - BACKGROUND: The causes underlying the phenomenon of simultaneous medullary (MTC) and papillary thyroid cancer (PTC) are unclear. METHODS: This study of 26 patients with simultaneous MTC and PTC aimed at clarifying clinical and histopathologic characteristics and trends of this unusual condition among MTC and PTC patients who were operated on at a tertiary referral center in Germany. RESULTS: A total of 26 patients revealed simultaneous MTC and PTC, yielding the following rates: 2.6% (26 patients) among all 1019 PTC patients, 2.6% (6 patients) among all 235 hereditary MTC patients, 4.1% (20 patients) among all 492 sporadic MTC patients, and 3.6% (26 patients) among all 727 MTC patients. Simultaneous MTC and PTC were consistently smaller than nonsimultaneous MTC (6 vs. 13 mm for hereditary MTC, P = .16; 12 vs. 23 mm for sporadic MTC, P = .009; and 11 vs. 20 mm for any MTC, P = .008) and PTC (8 vs. 20 mm, P < .001). Simultaneous MTC and PTC increased among MTC and PTC patients over time: from 0% to 4.3% (PTC), from 0% to 4.6% (hereditary MTC), from 0% to 8.1% (sporadic MTC), and from 0% to 7.0% (any MTC). For sporadic MTC, these rates virtually doubled every 5 years. Of 6 patients with simultaneous hereditary MTC and PTC, 5 revealed late-onset REarranged during Transfection (RET) mutations (1 L790F carrier; 2 V804L and 2 S891A carriers). CONCLUSIONS: Greater pathologic scrutiny, in addition to environmental changes, explains the surge of simultaneous MTC and PTC in Germany. More data are needed from additional geographic areas and populations to delineate individual contributions of these factors. PMID- 21626081 TI - Office ductoscopy for surgical selection in women with pathologic nipple discharge. AB - BACKGROUND: Pathologic nipple discharge (PND) is diagnosed clinically and managed by diagnostic duct excision (DDE). Mammary ductoscopy in the office setting may change this standard. We performed a prospective study to assess the utility of office ductoscopy for surgical selection in women with nipple discharge. METHODS: Women with nipple discharge meeting at least 2 of 3 criteria of PND (spontaneous, single duct, bloody or serous) underwent office ductoscopy. Those showing papillomatous lesions underwent DDE in the operating room (surgical group, n = 38); if no lesion was present, women were followed clinically (observation group, n = 21). RESULTS: A papillomatous lesion was identified in 79% of women with 3 criteria PND and in 21% with 2 criteria (P = .001). DDE yielded a proliferative lesion in 35 of 38 women (92%). Of the 38, 27 (71%) had papillomata, 2 (5%) had florid hyperplasia, and 6 (16%) had ductal carcinoma in situ (DCIS) on final pathology. Also, 11 women with papilloma and 1 with DCIS presented with 2 criteria PND. Ductoscopy findings were a better predictor of the presence of intraductal neoplasia (area under curve [AUC] 0.9, 95% confidence interval [95% CI] 0.8-0.98) compared with 3-criteria PND (AUC 0.7, 95% CI 0.6-0.8). The 21 women in the observation group did not develop signs of malignancy or need biopsy during a 48-month follow-up period. CONCLUSIONS: Our findings suggest that office ductoscopy provides accurate surgical selection of women with nipple discharge and should be considered for women with 2 criteria of PND, and those with negative ductoscopy can be safely observed. These findings need confirmation in a larger study with longer follow-up. PMID- 21626082 TI - Circulating angiogenic factors, tumor cells, and outcome after resection of metastatic colorectal cancer--what does it mean? PMID- 21626083 TI - A prospective study of religiousness and psychological distress among female survivors of Hurricanes Katrina and Rita. AB - This prospective study examined the pathways by which religious involvement affected the post-disaster psychological functioning of women who survived Hurricanes Katrina and Rita. The participants were 386 low-income, predominantly Black, single mothers. The women were enrolled in the study before the hurricane, providing a rare opportunity to document changes in mental health from before to after the storm, and to assess the protective role of religious involvement over time. Results of structural equation modeling indicated that, controlling for level of exposure to the hurricanes, pre-disaster physical health, age, and number of children, pre-disaster religiousness predicted higher levels of post disaster (1) social resources and (2) optimism and sense of purpose. The latter, but not the former, was associated with better post-disaster psychological outcome. Mediation analysis confirmed the mediating role of optimism and sense of purpose. PMID- 21626084 TI - The test of time in school-based mentoring: the role of relationship duration and re-matching on academic outcomes. AB - The influence of match length and re-matching on the effectiveness of school based mentoring was studied in the context of a national, randomized study of 1,139 youth in Big Brothers Big Sisters programs. The sample included youth in grades four through nine from diverse racial and ethnic backgrounds. At the end of the year, youth in intact relationships showed significant academic improvement, while youth in matches that terminated prematurely showed no impact. Those who were re-matched after terminations showed negative impacts. Youth, mentor, and program characteristics associated with having an intact match were examined. Youth with high levels of baseline stress and those matched with college student mentors were likely to be in matches that terminated prematurely, while rejection-sensitive youth and mentors who had previous mentoring experience were more likely to be in intact relationships. Implications for research and practice are discussed. PMID- 21626085 TI - Partial tibial nerve transfer to the tibialis anterior motor branch to treat peroneal nerve injury after knee trauma. AB - BACKGROUND: Injuries to the deep peroneal nerve result in tibialis anterior muscle paralysis and associated loss of ankle dorsiflexion. Nerve grafting of peroneal nerve injuries has led to poor function; therefore, tendon transfers and ankle-foot orthotics have been the standard treatment for foot drop. QUESTIONS/PURPOSES: We (1) describe an alternative surgical technique to obtain ankle dorsiflexion by partial tibial nerve transfer to the motor branch of the tibialis anterior muscle; (2) evaluate ankle dorsiflexion strength using British Medical Research Council grading after nerve transfer; and (3) qualitatively determine factors that influence functional success of surgery. METHODS: We retrospectively reviewed 11 patients treated with partial tibial nerve transfers after peroneal nerve injury. Pre- and postoperative motor strength was measured. Patients completed questionnaires regarding pre- and postoperative gait and disability. RESULTS: One patient regained Grade 4 ankle dorsiflexion, three patients regained Grade 3, one patient regained Grade 2, and two patients regained Grade 1 ankle dorsiflexion. Four patients did not regain any muscle activity. Clinically apparent motor recovery occurred an average 7.6 months postoperatively. A majority of patients (nine) could walk and participate in activities. Seven patients did not wear ankle-foot orthotics and four patients did not limp. The donor deficits included weak toe flexion (two patients) and reduced calf circumference (seven patients). CONCLUSION: Our observations suggest nerve transfers to the deep peroneal nerve provide inconsistent ankle dorsiflexion strength, possibly related to the mechanism of peroneal nerve injury or delays in surgery. Despite variable strength, four patients achieved M3 or greater motor recovery, which enabled them to walk without assistive devices. LEVEL OF EVIDENCE: Level IV, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence. PMID- 21626086 TI - Forensic validation of the PowerPlex(r) ESI 16 STR Multiplex and comparison of performance with AmpFlSTR(r) SGM Plus(r). AB - We describe the forensic validation of Promega's PowerPlex(r) European Standard Investigator 16 (ESI 16) multiplex kit and compare results generated with the AmpFlSTR(r) SGM Plus(r) (SGM+) multiplex. ESI 16 combines the loci contained within the SGM+ multiplex with five additional loci: D2S441, D10S1248, D22S1045, D1S1656, and D12S391. A relative reduction in amplicon size of the SGM+ loci facilitates an increased robustness and amplification success of these amplicons with degraded DNA samples. Tests performed herein supplement ESI 16 data published previously with sensitivity, profile quality, mock casework, inhibitor and mixture study data collected in our laboratories in alignment with our internal technical and quality guidelines and those issued by the Scientific Working Group on DNA Analysis Methods (SWGDAM), the DNA Advisory Board (DAB) and the DNA working group (DNAWG) of the European Network of Forensic Science Institutes (ENFSI). Full profiles were routinely generated from a fully heterozygous single source DNA template using 62.5 pg for ESI 16 and 500 pg for SGM+. This increase in sensitivity has a consequent effect on mixture analyses and the detection of minor mixture components. The improved PCR chemistry confers enhanced tolerance to high levels of laboratory prepared inhibitors compared with SGM+ results. In summary, our results demonstrate that the ESI 16 multiplex kit is more robust and sensitive compared with SGM+ and will be a suitable replacement system for the analysis of forensic DNA samples providing compliance with the European standard set of STR loci. PMID- 21626087 TI - Potential forensic application of DNA methylation profiling to body fluid identification. AB - DNA analysis of various body fluid stains at crime scenes facilitates the identification of individuals but does not currently determine the type and origin of the biological material. Recent advances in whole genome epigenetic analysis indicate that chromosome pieces called tDMRs (tissue-specific differentially methylated regions) show different DNA methylation profiles according to the type of cell or tissue. We examined the potential of tissue specific differential DNA methylation for body fluid identification. Five tDMRs for the genes DACT1, USP49, HOXA4, PFN3, and PRMT2 were selected, and DNA methylation profiles for these tDMRs were produced by bisulfite sequencing using pooled DNA from blood, saliva, semen, menstrual blood, and vaginal fluid. The tDMRs for DACT1 and USP49 showed semen-specific hypomethylation, and the tDMRs for HOXA4, PFN3, and PRMT2 displayed varying degrees of methylation according to the type of body fluid. Preliminary tests using methylation-specific PCR for the DACT1 and USP49 tDMRs showed that these two markers could be used successfully to identify semen samples including sperm cells. Body fluid-specific differential DNA methylation may be a promising indicator for body fluid identification. Because DNA methylation profiling uses the same biological source of DNA for individual identification profiling, the determination of more body fluid specific tDMRs and the development of convenient tDMR analysis methods will facilitate the broad implementation of body fluid identification in forensic casework. PMID- 21626088 TI - How music-inspired weeping can help terminally ill patients. AB - Music's power to improve the 'human condition' has been acknowledged since ancient times. Something as counter-intuitive as weeping in response to music can ameliorate suffering for a time even for terminally ill patients. Several benefits-including catharsis, communication, and experiencing vitality-can be associated with grieving in response to "sad" music. In addressing the potential rewards of such an activity for terminally ill patients, this author combines concepts from philosopher Jerrold R. Levinson's article, entitled "Music and Negative Emotion," an illustration from a major motion picture, and supporting research from medical reports and aesthetic writings. Carefully offering this experience is recommended for patients who retain the capacity to express preference. PMID- 21626089 TI - Hip bone strength indices in overweight and control adolescent boys. AB - The influence of being overweight on bone strength in adolescents remains controversial. The aim of this study was to compare hip bone strength indices in overweight and control adolescent boys using hip structure analysis (HSA). This study included 25 overweight adolescent boys [body mass index (BMI) >25 kg/m(2)] and 31 maturation-matched controls (BMI <25 kg/m(2)). Body composition and bone mineral density were assessed by dual-energy X-ray absorptiometry (DXA). To evaluate hip bone strength, DXA scans were analyzed at femoral neck, intertrochanteric, and femoral shaft by the HSA program. Cross-sectional area (CSA), an index of axial compression strength, section modulus (Z), an index of bending strength, cross-sectional moment of inertia (CSMI), cortical thickness (CT), and buckling ratio (BR) were measured from bone mass profiles. Body weight, lean mass, fat mass, and BMI were higher in overweight boys compared to controls (P < 0.001). CSA, CSMI, and Z of the three sites (femoral neck, intertrochanteric, and femoral shaft) were higher in overweight boys compared to controls (P < 0.01). BR was not significantly different between the two groups at the three sites. After adjustment for either body weight, BMI, or fat mass, using a one-way analysis of covariance, there were no differences between the two groups regarding the HSA variables (CSA, Z, CSMI, CT, and BR). After adjusting for lean mass, overweight boys displayed higher values of femoral shaft CSA, CSMI, and Z in comparison to controls (P < 0.05). In conclusion, this study suggests that overweight adolescent boys have greater indices of bone axial and bending strength in comparison to controls at the femoral neck, the intertrochanteric, and the femoral shaft. PMID- 21626090 TI - Regadenoson is a safe and well-tolerated pharmacological stress agent for myocardial perfusion imaging in post-heart transplant patients. AB - BACKGROUND: The safety and tolerability of regadenoson (REG), a newer adenosine A(2a) receptor agonist, has not been tested in orthotopic heart transplant (OHT) patients. METHODS: Retrospective review of a tertiary care center experience of OHT patients who underwent a REG single-photon emission computed tomography (SPECT) study as part of the work-up for cardiac allograft vasculopathy. The control group included those same patients who had prior adenosine-based SPECT. RESULTS: A total of 40 patients met the above criteria. Mean time from OHT to adenosine-SPECT and REG-SPECT was 8.2 +/- 4.8 years vs 9.8 +/- 4.5 years, respectively (P < .001). Both vasodilators had similar side effect profiles (P = .10), produced significant heart rate acceleration and asymptomatic hypotension (P < .001). There were no episodes of bradycardia and/or AV block with REG. Despite adjustment for medication status, adenosine was still associated with more conduction abnormalities (8 vs 1 event with REG, P = .02) including five episodes of 2nd degree AV block (Mobitz type II) and three episodes of sinus pause. CONCLUSION: This is the first reported use of REG in OHT patients. REG appears to be safe and well tolerated without significant cardiovascular adverse events. PMID- 21626091 TI - Incremental prognostic value of coronary flow reserve assessed with single-photon emission computed tomography. AB - BACKGROUND: We assessed the prognostic value of coronary flow reserve (CFR) estimated by single-photon emission computed tomography (SPECT) in patients with suspected myocardial ischemia. METHODS AND RESULTS: Myocardial perfusion and CFR were assessed in 106 patients using dipyridamole/rest Tc-99m sestamibi SPECT and follow-up was obtained in 103 (97%) patients. Four early revascularized patients were excluded and 99 were assigned to normal (summed stress score <3) vs abnormal myocardial perfusion and to normal (>=2.0) vs abnormal CFR. During the follow-up (5.8 +/- 2.1 years), 28 patients experienced a cardiac event (cardiac death, nonfatal myocardial infarction, and late revascularization). Abnormal perfusion (P < .01) and abnormal CFR (P < .05) were independent predictors of cardiac events at Cox proportional hazard regression analysis. Also in patients with normal perfusion, abnormal CFR was associated with a higher annual event rate compared with normal CFR (5.2% vs 0.7%; P < .05). CFR data improved the prognostic power of the model including clinical and myocardial perfusion data increasing the global chi-square from 18.6 to 22.8 (P < .05). Finally, at parametric survival analysis, in patients with normal perfusion the time to achieve >=2% risk of events was >60 months in those with normal and <12 months in those with abnormal CFR. CONCLUSIONS: Myocardial perfusion findings and CFR at SPECT imaging are both independent predictors of cardiac events. Estimated CFR provides incremental prognostic information over those obtained from clinical and myocardial perfusion data, particularly in patients with normal perfusion findings. PMID- 21626092 TI - Absence of repellents in Ustilago maydis induces genes encoding small secreted proteins. AB - The rep1 gene of the maize pathogen Ustilago maydis encodes a pre-pro-protein that is processed in the secretory pathway into 11 peptides. These so-called repellents form amphipathic amyloid fibrils at the surface of aerial hyphae. A SG200 strain in which the rep1 gene is inactivated (?rep1 strain) is affected in aerial hyphae formation. We here assessed changes in global gene expression as a consequence of the inactivation of the rep1 gene. Microarray analysis revealed that only 31 genes in the ?rep1 SG200 strain had a fold change in expression of >=2. Twenty-two of these genes were up-regulated and half of them encode small secreted proteins (SSPs) with unknown functions. Seven of the SSP genes and two other genes that are over-expressed in the ?rep1 SG200 strain encode proteins that can be classified as secreted cysteine-rich proteins (SCRPs). Interestingly, most of the SCRPs are predicted to form amyloids. The SCRP gene um00792 showed the highest up-regulation in the ?rep1 strain. Using GFP as a reporter, it was shown that this gene is over-expressed in the layer of hyphae at the medium-air interface. Taken together, it is concluded that inactivation of rep1 hardly affects the expression profile of U. maydis, despite the fact that the mutant strain has a strong reduced ability to form aerial hyphae. PMID- 21626093 TI - Development and validation of a scale measuring satisfaction with maternal and newborn health care following childbirth. AB - To demonstrate the validity and internal consistency of a multi-item scale measuring women's satisfaction with health care received in the weeks following childbirth for both themselves and their newborns. Data are from 1,154 women delivering healthy singletons or twins recruited for a randomized trial. Satisfaction with care items were selected from prior research, including the previously validated Primary Care Satisfaction Survey for Women (PCSSW) and studies of postpartum care. After randomly splitting the sample (1:1) for cross validation purposes, Exploratory (EFA) and Confirmatory factor analysis (CFA) on ordinal data using the WLSMV estimator available in the MPLUS statistical modeling program were conducted. A CFA was also conducted on available data at 2 weeks and 2 months after discharge in order to examine internal consistency at follow-up. A one factor model with 11 items was selected, with the main factor explaining 83% of total 11-item variation in the overall sample. The Cronbach's alpha for the final 11-item scale at baseline and follow-up time periods was 0.96. High correlations with overall trust in provider (Spearman rho = 0.78) and quality of healthcare (0.75) supported convergent validity. The baseline mean score was 47.9 with a standard deviation of 7.13 and a possible range of 11 (low) to 55 (high satisfaction). This validated scale is a new tool for measuring satisfaction with health care received during the postpartum period for mothers and their newborns. This tool will be useful in studies assessing quality of care and the outcomes of postpartum health care interventions, and it is the first tool to focus on care for the mother-baby unit. PMID- 21626094 TI - The potential contribution of marital-cohabitation status to racial, ethnic, and nativity differentials in birth outcomes in Texas. AB - Disparities in infant mortality by race/ethnicity and nativity are widely known. Patterns of marriage and union formation also vary by race, ethnicity and nativity and may contribute to disparities in birth outcomes. Using population level data, we build on previous research of race/ethnic disparities in birth outcomes by investigating the role of union status. Data come from the 2006 Birth Record from Texas Vital Statistics. The final sample size included 369,839 births to Texas women aged 18 and older. Birth outcomes were constructed from indicators of low birth weight and preterm birth. Logistic regression estimates odds of low birth weight and preterm birth by race/ethnicity and nativity and union status. Race/ethnicity/nativity and union status are significant and independent predictors of birth outcomes. US born Black and Mexican Origin mothers had higher odds of preterm birth and low birth weight babies compared to US born White mothers. Unmarried mothers had higher odds of adverse birth outcomes compared to married women. There was only modest support that the association between race/ethnicity/nativity status and birth outcomes could be explained by divergent patterns in union status. Though disparities in birth outcomes are persistent across race, ethnicity and nativity, the results suggest that union status at birth is a very weak factor in accounting for these disparities. Differing patterns in union status did not account for the Black-White and Mexican Origin White gaps in infant health outcomes. Additional research aimed at uncovering the processes that put these mothers and infants at higher risk is needed. PMID- 21626096 TI - Effects of hyperbaric oxygen on the expression of claudins after cerebral ischemia-reperfusion in rats. AB - The malfunction of tight junctions (TJs) between endothelial cells in the blood brain barrier (BBB) is the pathophysiological basis for cerebral ischemia reperfusion (IR) injury. Claudins, major molecular elements of the TJs, play a key role in the paracellular permeability of the BBB. Although several studies have demonstrated the impact of hyperbaric oxygenation (HBO) on boosting oxygen supply and reducing infarct size, its effect and underlying mechanism on the integrity of the BBB is unknown. To study the function of HBO on claudins and the permeability of the BBB, we replicated the animal model of local cerebral IR. Using Evans blue dye, permeability of the BBB was examined. Transmission electron microscopy (TEM), immunohistochemistry, western blot, and gelatin zymography were used to detect the integrity of the BBB, the expression of claudin-1 and claudin 5, and the activity of matrix metalloproteinases (MMPs) in brain microvessel endothelium. Our data indicate that compared with the sham-operated group, IR increased permeability of the BBB to Evans blue dye (P < 0.01), peaking at 4 h. The BBB ultrastructure was disrupted and the expression of claudin-5 and claudin 1 decreased (P < 0.01) in the 4 and 72 h IR group, respectively. Increased claudin-5 and claudin-1 expression and decreased permeability of the BBB were observed in the HBO + IR group (P < 0.01) via the suppression of MMP-2 and MMP-9, respectively. Our study provides direct evidence that HBO decreases the permeability of the BBB by reducing the enzymatic activity of MMPs and augmenting the expression of claudins at different stages in cerebral IR injury. PMID- 21626097 TI - Experience of a patient with an extracorporeal ventricular assist system who participated in a sleepover program. AB - A 19-year-old woman suffered fulminant myocarditis owing to a mycoplasma infection and was inserted with an intra-aortic balloon pump and a percutaneous cardiopulmonary support. Antibiotics and gamma globulin were administered, however, the patient's cardiac function did not recover, and the TOYOBO ventricular assist device (VAD) was implanted. She had rehabilitation training such as maintaining a standing position at the bedside and walking in the hospital, and a hospital outing program to a family restaurant was conducted two times with the VAD. The patient wished to attend the coming-of-age ceremony in Tachikawa city, which is 3 h away from our hospital by car. Therefore, we planned the program including a night stay at her home. The patient and her family fully understood the risks and wished to participate in the sleepover program. In preparing for the sleepover, the patient and her family learned to operate the VAD, and she was able to move to the lavatory and through the house with the help of only her family. A physician and a clinical engineer stayed at her house for infusion of antibiotics and management of sudden changes. There was no adverse event. In Japan, the community support of patients with VAD is not yet established, and we hope that our experience becomes a help to support return to society for patients with VAD. PMID- 21626098 TI - Continuous monitoring of glucose levels in the hepatic vein and systemic circulation during the Pringle maneuver in beagles. AB - Intraoperative continuous glucose monitoring revealed that liver ischemia/reperfusion causes a rapid and profound transition in glucose concentration. We hypothesized that the washout of the glucose stored in the liver leads to a rapid transition in blood glucose concentration. Six female beagles were studied. A portosystemic shunt was established, and the glucose levels in the jugular, hepatic, and portal veins were continuously monitored. All beagles were stabilized for 30 min, and, subsequently, the hepatic artery and portal vein were clamped (the Pringle maneuver). After 30 min of warm hepatic ischemia, the clamp was removed in order to initiate hepatic reperfusion. The endpoint of the experiment was 60 min after the onset of reperfusion. The glucose levels in the abovementioned veins were recorded continuously. The glucose level in the hepatic vein increased 10 min after the start of the Pringle maneuver and was significantly higher than that in the jugular vein and portal vein just before unclamping. The glucose level in the hepatic vein peaked at 2 min after unclamping and that in the portal and jugular veins started to increase after reperfusion. The glucose level in the hepatic vein was significantly higher than that in the jugular and portal veins between 9 min after clamping and 8 min after reperfusion. In conclusion, our study among beagles showed that glucose release from the hepatic vein and sinusoid leads to a rapid elevation in systemic blood glucose levels after liver ischemia/reperfusion. This knowledge might help in the development of new strategies for blood glucose management during hepatectomy. PMID- 21626099 TI - A novel counterpulsation mode of rotary left ventricular assist devices can enhance myocardial perfusion. AB - The effect of rotary left ventricular assist devices (LVADs) on myocardial perfusion has yet to be clearly elucidated, and several studies have shown decreased coronary flow under rotary LVAD support. We have developed a novel pump controller that can change its rotational speed (RS) in synchronization with the native cardiac cycle. The aim of our study was to evaluate the effect of counterpulse mode, which increases the RS in diastole, during coronary perfusion. Experiments were performed on ten adult goats. The EVAHEART LVAD was installed by the left ventricular uptake and the descending aortic return. Ascending aortic flow, pump flow, and coronary flow of the left main trunk were monitored. Coronary flow was compared under four conditions: circuit-clamp, continuous mode (constant pump speed), counterpulse mode (increased pump speed in diastole), and copulse mode (increased pump speed in systole). There were no significant baseline changes between these groups. In counterpulse mode, coronary flow increased significantly compared with that in continuous mode. The waveform analysis clearly revealed that counterpulse mode mainly resulted in increased diastolic coronary flow. In conclusion, counterpulse mode of rotary LVADs can enhance myocardial perfusion. This novel drive mode can provide great benefits to the patients with end-stage heart failure, especially those with ischemic etiology. PMID- 21626103 TI - Influence of insulin on glutamine synthetase in the Muller glial cells of retina. AB - Glutamine synthetase (GS), a Muller cell specific enzyme in the retina, is the key enzyme involve in glutamate metabolism. The goal of this study was to investigate the expression and regulation of GS by insulin in the cultured rat retinal Muller cells. Immunocytochemical and immunoblotting experiments showed that the cultured Muller cells express GS protein under normal cell culture conditions. Insulin treatments decreased the GS expression both in a time and dose dependent manner. Insulin also decreased the hydrocortisone induced GS expression. Furthermore, we investigated the expression and regulation of two other Muller cell specific enzymes known to be involved in glutamate metabolism, the mitochondrial branched chain aminotransferase (BCATm) and pyruvate carboxylase (PC). Immunoblotting experiments showed that Muller cells expressed both BCATm and PC. Treatments of cells with hydrocortisone or insulin did not influence the BCATm expression level. Hydrocortisone treatment of cells increased the PC expression but this induced expression was suppressed by insulin treatment. Muller cells expressed insulin receptor proteins (IRbeta and IRS-1) and insulin activation induced the phosphotyrosine level of insulin receptor proteins. Moreover, hydrocortisone did not influence the expression or activation of these receptor proteins. The data suggests that insulin modulates the GS synthesis and may influence glutamate metabolism in the cultured retinal Muller cells but not by influencing the insulin signaling pathway. PMID- 21626104 TI - Rebound enlargement of an ectopic cervical thymus mimicking relapse of lymphoblastic lymphoma. PMID- 21626105 TI - Autoimmune lymphoproliferative syndrome mimicking chronic active Epstein-Barr virus infection. AB - Chronic active Epstein-Barr virus infection (CAEBV) is defined as a systemic EBV associated lymphoproliferative disease characterized by fever, lymphadenopathy, and splenomegaly in apparently immunocompetent persons. Recent studies have revealed that EBV infects T or natural killer cells in most patients with CAEBV; the etiology of CAEBV, however, remains unknown. Autoimmune lymphoproliferative disorder (ALPS) is an inherited disorder associated with defects in apoptosis, and clinically characterized by lymphadenopathy, splenomegaly, hypergammaglobulinemia, and autoimmune disease. ALPS is most often associated with mutations in the FAS gene, which is an apoptosis-signaling receptor important for homeostasis of the immune system. Based on the clinical similarity between ALPS and CAEBV with respect to lymphoproliferation, we have examined the possibility of the co-occurrence of ALPS in patients with a diagnosis of CAEBV. In this study, we have identified FAS gene mutations in three Japanese patients with lymphadenopathy, hepatosplenomegaly, and unusual EBV infection, who were diagnosed with CAEBV. These observations, which indicate that the clinical development of ALPS may be associated with EBV infection, alert us to a potential diagnostic pitfall of CAEBV. PMID- 21626107 TI - Rebalancing the risks of Computed Tomography and Magnetic Resonance imaging. PMID- 21626106 TI - Utility of laboratory tests in B-CLL patients in different clinical stages. AB - The study objective was to analyse the utility of laboratory tests performed in 30 patients with B-cell chronic lymphocytic leukaemia (B-CLL) at different clinical stages. Laboratory tests included automated and microscopic assessment of peripheral blood and bone marrow counts as well as evaluation of leukaemic cells. Apart from the diagnostic and prognostic value of laboratory abnormalities such as clonal lymphocytosis with CD5+CD19+CD23+ phenotype, reduced erythrocyte parameters, thrombocytopenia or bone marrow infiltration by the neoplastic clone as well as low percentage of Gumprecht's shadows, low apoptotic activity of peripheral blood lymphocytes, and increased percentage of CD38- and ZAP-70 +/- cells markedly correlate with the stage of disease progression. These results seem to confirm the diagnostic and prognostic significance of these parameters determined in routine laboratory tests in B-CLL patients. PMID- 21626108 TI - Nrf2-dependent gene expression is affected by the proatherogenic apoE4 genotype studies in targeted gene replacement mice. AB - An apoE4 genotype is an important risk factor for cardiovascular and other chronic diseases. The higher cardiovascular disease risk of apoE4 carriers as compared to the apoE3 genotype has been mainly attributed to the differences in blood lipids between the two genotype subgroups. Recently, a potential protective role of the transcription factor Nrf2 in cardiovascular disease prevention has been suggested. In this study, we show that Nrf2-dependent gene expression is affected by the apoE genotype. ApoE4 vs. apoE3 mice exhibited lower hepatic Nrf2 nuclear protein levels. Furthermore, mRNA and protein levels of Nrf2 target genes including glutathione-S-transferase, heme oxygenase-1 and NAD(P)H dehydrogenase, quinone 1 were significantly lower in apoE4 as compared to apoE3 mice. Lower hepatic mRNA levels of phase II enzymes, as observed in apoE4 vs. apoE3 mice, were accompanied by higher mRNA levels of phase I enzymes including Cyp26a1 and Cyp3a16. Furthermore, miRNA-144, miRNA-125b, and miRNA-29a involved in Nrf2 signaling, inflammation, and regulation of phase I enzyme gene expression were affected by the apoE genotype. We provide first evidence that Nrf2 is differentially regulated in response to the apoE genotype. PMID- 21626109 TI - Response of denitrification genes nirS, nirK, and nosZ to irrigation water quality in a Chinese agricultural soil. AB - PURPOSE: Denitrification is an important biochemical process in global nitrogen cycle, with a potent greenhouse gas product N(2)O. Wastewater irrigation can result in the changes of soil properties and microbial communities of agricultural soils. The purpose of this study was to examine how the soil denitrification genes responded to different irrigation regimes. MATERIALS AND METHODS: Soil samples were collected from three rural districts of Beijing (China) with three different irrigation regimes: clean groundwater (CW), reclaimed water (RW), and wastewater (WW). The abundance and diversity of three denitrification microbial genes (nirS, nirK, and nosZ) were examined by real-time polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE) molecular approaches. RESULTS AND DISCUSSION: The abundance of nirS in the WW treatment was higher than that in the CW treatment, and no significant difference was found between the RW and CW or WW treatments. The abundance of nirK gene of the RW and WW treatments was higher than that of the CW treatment. There was no difference for nosZ gene among the three treatments. Correspondence analysis based on the DGGE profiles showed that there was no obvious difference in the nosZ gene composition, but nirS and nirK genes changed with different irrigation regimes. CONCLUSIONS: Irrigation with unclean water sources enhanced the soil NO (3) (-) content and changed the abundance and composition of soil denitrifiers, and different functional genes had different responses. Irrigation with unclean water sources increased the abundance of nirK gene and changed the community structures of nirS and nirK genes, while nosZ gene was relatively stable in the soil. These results could be helpful to explore the mechanisms of the variation of denitrification processes under long-term wastewater irrigation and partially explain the reason of more N(2)O output in the field with wastewater irrigation. PMID- 21626110 TI - Levels and congener profiles of particle-bound polybrominated dibenzo-p dioxins/furans (PBDD/Fs) in ambient air around Guangzhou, China. AB - The mean particulate polybrominated dibenzo-p-dioxins/furans (PBDD/Fs) levels for Guangzhou were in the range of 225-2244 fg/m(3) and showed the spatial trend of background < suburb areas < urban area < industrial area, suggesting the industrial and urban sources of PBDD/Fs. The most abundant congeners were 2,3,7,8 TBDF, 1,2,3,7,8-PeBDF and 2,3,4,7,8-PeBDF. The mean toxicity equivalency quantities (TEQs) of PBDD/Fs were in the range of 89.3-456 fg I-TEQ/m(3) for Guangzhou, which were several times higher than that of the background area (49.1 fg I-TEQ/m(3)). The rough inhalation risk evaluation showed that the residents in Guangzhou were in relatively higher exposure levels of PBDD/Fs. PMID- 21626111 TI - Toxicity prediction of dioxins and dioxins-like compounds based on the molecular fragments variable connectivity index. AB - The toxicity of 95 doxins and dioxin-like compounds was investigated by quantitative structure-activity relationship (QSAR) with the molecular fragments variable connectivity index (mfVCI). For each of the four homologues, the models have good fitting (R2 > 0.80) and predictive (Q2EXT > 0.80) ability. The models developed from more than one homologues are also satisfactory with R2 > 0.80 and Q2EXT > 0.77. The molecular fragments weights have the ability to diagnose the contribution of the molecular fragments to the toxicity of the compounds. The mfVCI may play an important role in the development of molecular descriptors in further QSAR research. PMID- 21626112 TI - Pilot investigation of perfluorinated compounds in river water, sediment, soil and fish in Tianjin, China. AB - Detectable PFCs could be found in all samples. Perfluorooctanoic acid (PFOA) was the major PFC in river water, while perfluorooctane sulfonate (PFOS) was dominant in sediment and were 17- to 153-fold higher than those in water. PFCs concentrations in soil were little higher than those in sediment. In fish muscles PFOS showed the highest concentrations. Generally, PFC concentrations in fish were in the following rank order: crucian carp > silver carp> common carp. We suggested that there may be constant and diffuse pollution sources in Tianjin. Point sources also appeared to make significant contribution in the present study. PMID- 21626113 TI - Biphasic bone substitute and fibrin sealant for treatment of benign bone tumours and tumour-like lesions. AB - PURPOSE: Bone defects resulting from tumour resection or curettage are most commonly reconstructed with autologous bone graft which is associated with limited availability and donor site morbidity. Recent research has focussed on synthetic biomaterials as bone graft substitutes. The aim of this study was to assess the safety and efficiency of a bone substitute as an alternative for autologous bone in the treatment of benign bone tumours and tumour-like lesions. METHODS: In the present study, a biphasic ceramic (60% HA and 40% beta-TCP) combined with a fibrin sealant was used to reconstruct defects in 51 patients after curettage of benign bone tumours or tumour-like lesions. Patient age ranged from eight to 68 years (mean 29.7), defect size from 2 cm(3) to 35 cm(3) (mean 12.1), and time of follow-up from one to 56 months (mean 22.7). RESULTS: Radiologic analysis showed complete bony defect consolidation in 50 of 51 patients after up to 56 months. No postoperative fractures were observed. Revision surgery had to be performed in one case. Histological analysis showed new bone formation and good biocompatibility and osseointegration of the implanted material. CONCLUSION: In summary, the biphasic ceramic in combination with fibrin sealant was proven an effective alternative to autologous bone grafts eliminating the risk of donor site morbidity for the patient. PMID- 21626114 TI - Nutlin-3 differentially modulates miRNA34a and miRNA181 versus miR26a and miR155 in p53 proficient and p53 deficient B chronic lymphocytic leukemia (B-CLL) samples. PMID- 21626115 TI - Two-stage model-based design of cancer phase I dose escalation trials: evaluation using the phase I program of barasertib (AZD1152). AB - INTRODUCTION: Modeling and simulation of pharmacokinetics and pharmacodynamics has previously been shown to be potentially useful in designing Phase I programs of novel anti-cancer agents that show hematological toxicity. In this analysis, a two-stage model-based trial design was evaluated retrospectively using data from the Phase I program with the aurora kinase inhibitor barasertib. METHODS: Data from two Phase I trials and four regimens were used (n = 79). Using barasertib hydroxy QPA plasma concentrations and neutrophil count data from only study 1A, a PKPD model was developed and subsequently used to predict the MTD and a safe starting dose for the other trials. RESULTS: The PKPD model based on data from the first study adequately described the time course of neutrophil count fluctuation. The two-stage model-based design provided safe starting doses for subsequent phase I trials for barasertib. Predicted safe starting dose levels were higher than those used in two subsequent trials, but lower than used in the other trial. DISCUSSION: The two-stage approach could have been applied safely to define starting doses for alternative dosing strategies with barasertib. The limited improvement in efficiency for the phase I program of barasertib may have been due to the fact that starting doses for the studied phase I trials were already nearly optimal. CONCLUSION: Application of the two-stage model-based trial design in Phase I programs with novel anti-cancer drugs that cause haematological toxicity is feasible, safe, and may lead to a reduction in the number of patient treated at sub-therapeutic dose-levels. PMID- 21626116 TI - What is the added value of combined core biopsy and fine needle aspiration in the diagnostic process of renal tumours? AB - PURPOSE: Non-diagnostic results still hinder the routine use of core biopsy (CB) and fine needle aspiration (FNA) in the diagnostic process of renal tumours. Furthermore, substantial interobserver variability has been reported. We assessed the added value of combining the results of CB and FNA by five pathologists in the ex vivo diagnosis of renal mass. METHODS: Two ex vivo core biopsies were taken followed by two FNA passes from extirpated tumours. All samples were evaluated by five blinded pathologists. A consensus diagnosis of the surgical specimen was the index for comparison. For each pathologist, the number of non diagnostic (non-conclusive or undetermined biology and failed biopsies), correct and incorrect scored cases of each technique was assessed. When a non-diagnostic CB or FNA had a correct diagnostic counterpart, this was considered as of added value. RESULTS: Of the 57 assessed tumours, 53 were malignant. CB was non diagnostic in 4-10 cases (7-17.5%). FNA established the correct diagnosis in 1-7 of these cases. FNA was non-diagnostic in 2-6 cases (3.5-10.5%), and the counterpart CB established the correct diagnosis in 1-6 of these cases. For the 5 pathologists, accuracy of CB and FNA varied between 82.5-93% and 89.5-96.5%, respectively. Combination of both types of biopsy resulted in 55-57 correct results (accuracy 96.5-100%), i.e., an increase in accuracy of 3.5-14%. CONCLUSION: Combining the result of CB and FNA in renal mass biopsy leads to a higher diagnostic accuracy. Recommendations on which technique used should be adapted to local expertise and logistic possibilities. PMID- 21626117 TI - Extracellular matrix remodeling in the human gubernaculum during fetal testicular descent and in cryptorchidic children. AB - PURPOSE: We evaluated extracellular matrix remodeling in human fetal and cryptorchidic gubernacula. METHODS: Gubernacula were obtained from 40 normal human fetuses aged 15-29 weeks postconception (WPC) and from 39 children aged 1.3 10 years who had been submitted to orchiopexy. Total collagen and glycosaminoglycan (GAG) concentrations were assessed as MUg hydroxyproline and MUg hexuronic acid per mg dry gubernacular tissue, respectively, and proportions of sulfated GAG were determined by agarose gel electrophoresis. RESULTS: Fetal age correlated with collagen (r = 0.77, P < 0.001) and GAG (r = -0.83, P < 0.01) concentrations, which varied from 10 to 50 MUg/mg and 7.1-2.5 MUg/mg, respectively. Collagen and GAG concentrations in cryptorchidic gubernacula varied from 80.0 to 120.0 MUg/mg and from 0.8 to 1.0 MUg/mg, respectively. These values did not correlate with patient's age, but when the testis was located more proximally, collagen content was lower. At 15-18 WPC, GAG consisted of 57.3% +/- 13.5% chondroitin sulfate and 28.2% +/- 10.1% dermatan sulfate, and at 25-28 WPC, 42.7% +/- 8.7% and 71.8% +/- 11.3%, respectively. GAG in postnatal gubernacula consisted mostly of dermatan sulfate. CONCLUSIONS: From the 15th to the 29th WPC, the extracellular matrix of the gubernaculum undergoes extensive remodeling and this may contribute to testicular descent. Cryptorchidic gubernacula are markedly more fibrous than the corresponding fetal tissue, change little after birth, and have a lower collagen concentration when the undescended testis is abdominal in position. PMID- 21626118 TI - Can the intestinal dysmotility of critical illness be differentiated from postoperative ileus? AB - Gastrointestinal dysmotility is commonly noted in the intensive care unit and postoperative settings. Characterized by delayed passage of stool and flatus, nausea, vomiting, and abdominal distention, the condition is associated with nutritional deficiencies, risk of aspiration, and considerable allocation of health care resources. Knowledge of gastrointestinal function in health and illness continues to expand. While the factors that precipitate ileus differ between postoperative and critically ill patients, the two clinical scenarios seem to have similar mechanisms and share many of the same pathophysiologic patterns. By reviewing and comparing the literature on the respective mechanisms and contributing factors generated in these separate clinical settings, a common more comprehensive management strategy may be derived with the potential for newer innovative therapeutic options. PMID- 21626119 TI - Helicobacter pylori: is it a complete villain? PMID- 21626120 TI - Obstructive sleep apnea in Treacher Collins syndrome. AB - The aim of the present study was to investigate the prevalence of obstructive sleep apnea syndrome (OSAS) among the Norwegian population with Treacher Collins syndrome (TCS). A secondary aim was to establish whether TCS phenotype severity is associated with OSAS severity. A prospective case study design was used. Individuals who were 5 years old and above with a known diagnosis of TCS in Norway were invited to participate in a study. The study included genetic testing, medical and dental examinations and polysomnography. All participants demonstrated disturbed respiration during sleep; 18/19 met the diagnostic criteria for OSAS. Subjectively evaluated snoring was not a reliable predictor of OSAS. We found no significant association between TCS phenotype severity and the severity of OSAS. OSAS is common in TCS, but there is no association with the phenotype severity. Individuals diagnosed with TCS must undergo sleep studies to identify the presence of OSAS. PMID- 21626121 TI - High-dosage betahistine dihydrochloride between 288 and 480 mg/day in patients with severe Meniere's disease: a case series. AB - The objective of this study was to evaluate the clinical benefit and the side effects of high dosages of betahistine dihydrochloride (288-480 mg/day) in patients with severe Meniere's disease (MD). In this case series 11 patients with MD who had not responded sufficiently to a dosage of 144 mg/day of betahistine dihydrochloride were treated on an individual basis with daily dosages between 288 and 480 mg of betahistine dihydrochloride. The number of attacks per month and the side effects were monitored. Non-parametric tests were used for statistical analysis. As a result, the frequency and the severity of vertigo were significantly reduced in all patients. The side effects were mild, self-limiting, and did not require any change in the treatment strategy. Despite the considerable limitations of an observational study--in particular in MD--high dosages of betahistine dihydrochloride between 288 and 480 mg/day seem to be effective in patients who do not sufficiently respond to lower dosages. Moreover, such dosages are well tolerated. PMID- 21626122 TI - Rarely reported ocular complications following surgery to correct chronic suppurative otitis media. AB - The objective of this study was to describe the rarely reported ocular complications following surgery to correct chronic suppurative otitis media. A retrospective analysis was made of 2,318 cases of chronic suppurative otitis media treated with surgery between January 2004 and December 2009 at West China Hospital, Sichuan University, a medical center in Western China. Ninety-two cases of ocular complications were identified, giving an incidence of 3.97%. In 63 of the 92 cases (68.48%), the patients complained of blurred vision and the condition was classified as grade 1. In 24 cases (26.09%), the patients complained of mild periorbital edema and the condition was classified as grade 2. Four cases (4.35%) were classified as grade 3 because of severe edema with erythema on eyelids. One case (1.09%) was classified as grade 4 because of periorbital ecchymosis in addition to edema. The clinical features and the possible etiology were analyzed and are discussed. Ocular complications following chronic suppurative otitis media surgery are rare. Preauricular incision, disturbance of venous and lymphatic drainage, and preseptal cellulitis were the possible etiological factors. Eye injury during surgery and use of an anticholinergic agent might be risk factors for blurred vision after this surgery. PMID- 21626123 TI - Does intratympanic gadolinium injection predict efficacy of gentamicin partial chemolabyrinthectomy in Meniere's disease patients? AB - Using actual diagnostic criteria, the diagnosis of certain Meniere's disease remains impossible during life without histopathologic confirmation. Assessing the value of a diagnostic test is difficult due to the lack of a gold standard. Recent studies reported on the use of MRI after intratympanic gadolinium injection to demonstrate endolymphatic hydrops in vivo. We evaluate whether MRI after intratympanic gadolinium administration is useful for predicting the effect and outcome of intratympanic gentamicin therapy. The correlation between transtympanic electrocochleographic (TT-ECoG) results and hydrops grade on MRI images is also investigated. Twelve definite Meniere's disease patients with incapacitating vertigo attacks, not responding to drug and behavioral treatment, were selected for partial chemolabyrinthectomy with intratympanic gentamicin. All patients underwent transtympanic electrocochleography followed by surgical middle ear inspection, partial chemolabyrinthectomy (gentamicin solution 40 mg/ml applied during 60 min) and intratympanic gadolinium injection with clear exposure of the round window membrane. The MR images were reviewed and a hydrops grade was assigned. Correlation between the hydrops grade and the electrocochleographic data was assessed. Only 5 of 12 patients showed gadolinium enhancement in the inner ear. However, 6 of the 7 patients that did not show postoperative intracochlear or intralabyrinthine gadolinium distribution did report the clinical improvement after intratympanic gentamicin therapy. Hydrops grade correlated with the result of transtympanic electrocochleography in four of five cases that showed gadolinium enhancement. We conclude that the use of intratympanic gadolinium has no added value in predicting the clinical outcome of intratympanic gentamicin application. However, based on these data, a correlation between the result of TT-ECoG and hydrops grading on MRI images can be suggested. PMID- 21626124 TI - Multilevel radiofrequency ablation for snoring and OSAHS patients therapy: long term outcomes. AB - Our objective is to evaluate the outcomes of the Radio-Frequency (RF) energy for tissue thermo-ablation therapy in sleep-disordered breathing patients and retrospective evaluation of the RF therapy after a 5-year follow-up period, in terms of snoring and apnea reduction. From June 1999 to June 2009, we enrolled patients suffering from simple snoring and patients with obstructive apnea hypopnoea syndrome (OSAHS). A visual analog scale (VAS) questionnaire was used to evaluate the level of snoring and was filled out in short- and long-term periods, whereas in OSAHS patients an unattended polysomnography was performed before and after a minimum of 6 months from the last RF therapy treatment session. The presence of post-operative pain was assessed by means of a specific VAS. Results stated that 187/250 patients finished the RF therapy. In the simple snoring group, mean snoring VAS decreased from 7.48 to 3.7 (P < 0.0001). In the post operative snoring group, mean snoring VAS decreased from 7.6 to 3.6 (P < 0.0001). In the mild-to-moderate grade OSAHS group, AHI decreased from a mean value of 18.1 to a mean value of 12.9 (P < 0.0001). Furthermore, we recorded a mean post operative pain VAS of one in each group of patients. Our results suggest an important role of RF therapy in the improvement of snoring solution, but not for a significant AHI reduction. Level of evidence 2c. PMID- 21626125 TI - SIV/macaque model of HIV infection in cocaine users: minimal effects of cocaine on behavior, virus replication, and CNS inflammation. AB - Studies of the effects of drugs of abuse on HIV immune status, disease progression, and neuroAIDS have produced conflicting data and have not definitively shown whether this combination promotes cognitive impairment or disease progression. Using a consistent SIV-macaque model, we investigated the effects of cocaine on behavior, virologic parameters, and CNS inflammation. Macaques received either vehicle or chronic administration of behaviorally active doses of cocaine (1.7 or 3.2 mg/kg/day). Chronic cocaine administration reduced CD8+ T cell counts during acute and late stage infection but had no effect on CD4+ T cell counts. Low-dose cocaine-treated animals had lower CSF vRNA levels late in infection, but cocaine did not alter plasma viral load or vRNA or protein in brain. There were no differences in CSF CCL-2 or interleukin (IL)-6 levels or severity of encephalitis in cocaine-treated as compared to vehicle-treated macaques. There were no differences in brain inflammation or neurodegeneration markers, as determined by interferon (IFN)-beta, MxA, CCL2, IL-6, TNFalpha, IFNgamma, and indolamine 2,3-deoxygenase mRNA levels. APP levels also were not altered. The executive function of inhibitory control was not impaired in cocaine treated or control animals following SIV infection. However, animals receiving 3.2 mg/kg/day cocaine performed more slowly in a bimanual motor test. Thus, chronic administration of cocaine produced only minor changes in behavior, encephalitis severity, CNS inflammation/neurodegeneration, and virus replication in SIV-infected pigtailed macaques, suggesting that cocaine would have only modest effects on the progression of neuroAIDS in HIV-infected individuals. PMID- 21626126 TI - Arachidonic acid accumulates in the stromal macrophages during thymus involution in diabetes. AB - Diabetes is a debilitating disease with chronic evolution that affects many tissues and organs over its course. Thymus is an organ that is affected early after the onset of diabetes, gradually involuting until it loses most of its thymocyte populations. We show evidence of accumulating free fatty acids with generation of eicosanoids in the diabetic thymus and we present a possible mechanism for the involution of the organ during the disease. Young rats were injected with streptozotocin and their thymuses examined for cell death by flow cytometry and TUNEL reaction. Accumulation of lipids in the diabetic thymus was investigated by histology and electron microscopy. The identity and quantitation of accumulating lipids was done with gas chromatography-mass spectrometry and liquid chromatography. The expression and dynamics of the enzymes were monitored via immunohistochemistry. Diabetes causes thymus involution by elevating the thymocyte apoptosis. Exposure of thymocytes to elevated concentration of glucose causes apoptosis. After the onset of diabetes, there is a gradual accumulation of free fatty acids in the stromal macrophages including arachidonic acid, the substrate for eicosanoids. The eicosanoids do not cause thymocyte apoptosis but administration of a cyclooxygenase inhibitor reduces the staining for ED1, a macrophage marker whose intensity correlates with phagocytic activity. Diabetes causes thymus involution that is accompanied by accumulation of free fatty acids in the thymic macrophages. Excess glucose is able to induce thymocyte apoptosis but eicosanoids are involved in the chemoattraction of macrophage to remove the dead thymocytes. PMID- 21626127 TI - Segregation during cleavage in the mammalian embryo? A critical comparison of whole-mount/CLSM and section immunohistochemistry casts doubts on segregation of axis-relevant leptin domains in the rabbit. AB - Segregation of certain cytoplasmic molecules during cleavage and blastocyst formation that was previously reported to occur in the human and the mouse (Antczak and Van Blerkom Mol Hum Reprod 3:1067-1086, 1997; Antczak and Van Blerkom Hum Reprod 14:429-447, 1999) has been reinvestigated in the rabbit model. Additional methodology was used and two approaches were compared: (1) whole-mount immunohistochemistry followed by confocal laser scanning microscopy (WM-IHC/CLSM) versus (2) IHC performed on histological sections of resin-embedded material (S IHC). This study concentrates on leptin and cytoskeletal proteins (actin and cytokeratins). With S-IHC, leptin was localized predominantly on the surface of blastomeres which is facing the perivitelline space, and in the extracellular embryonic coats, without any polar asymmetry being detectable along (presumptive) embryonic axes. A polar distribution of leptin with a pattern that could be interpreted as predictive of the prospective embryonic-abembryonic axis was seen only with WM-IHC/CLSM, not with S-IHC, although the latter gave excellent resolution. With both techniques, no differences between blastomeres were detected with respect to actin and cytokeratin patterns, an increased expression of cytokeratin in trophoblast cells occurring no earlier than at blastocyst formation. Artifacts that can occur with the two methodological approaches are critically discussed, as is the possible significance of the findings for theories on the differentiation of trophoblast versus embryoblast and on axis formation in early mammalian development. It is concluded that these data call for cautioning when studying distribution patterns of diffusible molecules with WM-IHC/CLSM technology, whereas patterns obtained with S-IHC are more reliable. Specifically these data cast doubts on previous claims that leptin IHC would allow to monitor cytoplasmic domain segregation occurring during cleavage as an element of early embryonic pattern/axis formation. PMID- 21626128 TI - Eye movements as a gatekeeper for memorization: evidence for the persistence of attentional sets in visual memory search. AB - Attention is known to serve multiple goals, including the selection of information for further perceptual analysis (selection for perception) and for goal-directed behavior (selection for action). Here, we study the role of overt attention (i.e., eye movements) as a gatekeeper for memorization processes (selection for memorization). Subjects memorized complex multidimensional stimulus displays and subsequently indicated whether a specific (probe) item was present. In Experiment 1 we utilized an incidental learning setting where in the beginning only a subset of display stimuli was relevant, whereas in a transfer block all stimuli were possible probe items. In Experiment 2, we used an explicit learning setting within a between-group design. Response times and gaze patterns indicated that subjects learned to ignore irrelevant stimuli while forming memory representations. The findings suggest that complex feature binding processes in peripheral vision may serve to guide overt selective attention, which eventually contributes to filtering out irrelevant information even in highly complex environments. Gaze patterns suggested that attentional control settings persisted even when they were no longer required. PMID- 21626129 TI - Response to modeling and notation of DEA with strong and weak disposable outputs. AB - In this brief communiction we respond to comments on our earlier publication, which contained some notational errors. PMID- 21626130 TI - Clinical cause of presumed acute tubular necrosis requiring renal replacement therapy and outcome of critically ill patients: post hoc analysis of a prospective 7-year cohort study. AB - BACKGROUND: Acute kidney injury (AKI) secondary to acute tubular necrosis (ATN) is common in critically ill patients, and causes significant morbidity and mortality. The underlying etiology of ATN can be divided into pure ischemic, pure nephrotoxic, and mixed causes. This post-hoc analysis of a prospective cohort study aimed to investigate whether the cause (pure vs. mixed) of ATN affects the short- and long-term outcome of critically-ill patients. METHODS: A total of 425 critically-ill patients with AKI secondary to clinically diagnosed ATN were divided into three groups according to the cause of ATN. Of these patients, 215 had mixed ATN, 203 had pure ischemic ATN, and seven had pure nephrotoxic ATN. All patients had one episode of AKI only. No patient had pre-existing chronic kidney disease. Patients were followed throughout their hospital stay (mortality rate, recovery of renal function at discharge) and up to 7 years thereafter. RESULTS: The three patient groups differed in their demographic and clinical characteristics. The in-hospital mortality rates were 55% in the presumably mixed cause ATN group, 39% in the pure ischemic group, and 29% in the pure nephrotoxic group. Complete renal recovery at discharge was documented in five out of five surviving patients with nephrotoxic ATN (100%) and in 92 out of 124 surviving patients with pure ischemic ATN (74%), but only in 29 out of 97 patients with mixed ATN (30%). None of the surviving patients was lost during the 7-year follow up. At the end of the observation period, 60% of the survivors of pure ATN, compared with 22% of the survivors of mixed ATN, were alive. After 7 years, 6% of the living patients with pure ATN had mild-to-moderate chronic kidney disease, whereas 38% of the mixed group patients had advanced CKD or end-stage renal disease. CONCLUSIONS: The cause of presumed ATN has a profound impact on short- as well as long-term outcomes of critically-ill patients with AKI requiring renal replacement therapy. The challenge for intensivists is to avoid further injury to the kidneys of these patients. PMID- 21626131 TI - Size of the transrectal ultrasound probe makes no difference in pain perception during TRUS-Bx under adequate local anesthesia. AB - INTRODUCTION AND OBJECTIVES: In the great majority of the cases, transrectal ultrasound guided prostate biopsy (TRUS-Bx) is the definitive step in the diagnosis of prostate cancer (CaP). Although this procedure is well tolerated by most patients, it can result in considerable discomfort that can effectively be overcome with local injection anesthesia. In this study, we evaluated the effect of the size (i.e., circumference) of the transrectal probe on pain during TRUS Bx. MATERIAL AND METHODS: One hundred and seventy eligible patients who had elevated total prostate specific antigen (tPSA) and/or abnormal digital rectal examination (DRE) were included in this study. Patients (pts.) were divided into three TRUS-Bx groups; Group I: 60 pts. underwent TRUS-Bx with newer B-K Type 8808 probe (circumference 58 mm) under injectable periprostatic anesthesia, Group II: 60 pts. underwent TRUS-Bx with B-K Type 8551 probe (circumference 74 mm) under injectable periprostatic anesthesia, and Group III: 50 pts. underwent TRUS-Bx with B-K Type 8551 probe (circumference 74 mm) without local anesthesia. Periprostatic injection anesthesia was performed with 10 cc, 1% lidocaine (5 cc on each side) 10 min before TRUS-Bx. Pain was assessed using a 10-point modified visual analog scale (VAS) 15 min after the biopsy procedure. RESULTS: Three groups were homogeneous with respect to age and tPSA, and no statistically significant difference was observed in terms of mean biopsy duration between the 3 groups. Most of the patients experienced no pain to slight pain in Groups I and II, but 66% of the patients had more than moderate pain (VAS >= 5) in Group III with mean VAS score statistically higher than the other two groups (Group I vs. III, P = 0.0001; Group II vs. III, P = 0.0001). Mean VAS score was not statistically different between Group I and II (P = 0.126). No statistically significant difference in VAS pain perception was observed between different age categories within the Group I, II, and III. CONCLUSION: In the absence of injectable local anesthesia, larger probe (74 mm) results in much higher VAS pain perception than same size and smaller (58 mm) probe used under injectable local anesthesia. However, under injectable local anesthesia, the size (circumference) of the transrectal probe (58 mm vs. 74 mm) does not result in any different pain perception during TRUS-Bx. PMID- 21626132 TI - Beneficial effect of aliskiren combined with olmesartan in reducing urinary protein excretion in patients with chronic kidney disease. AB - BACKGROUND: Blockade of the renin-angiotensin-aldosterone system is a therapeutic mainstay in patients with chronic kidney disease (CKD). However, the renoprotective effect of the novel direct renin inhibitor aliskiren is unknown. MATERIALS AND METHODS: We performed a prospective study in 10 CKD patients. All 10 patients with persistent proteinuria (urinary protein-to-creatinin ratio 0.3 3.5 g/g), despite good blood pressure control (<130/80 mmHg) with olmesartan, were started on 150 mg/day aliskiren. Clinical parameters were examined before and after 4, 8, 12, and 16 weeks of treatment. RESULTS: Urinary protein-to creatinine ratio significantly decreased by about 40% at 16 weeks from baseline (P = 0.0002), although estimated glomerular filtration rate and blood pressure did not change throughout the study period. Plasma renin activity also decreased significantly from baseline (P = 0.019), although plasma aldosterone concentration did not change. CONCLUSIONS: Aliskiren combined with olmesartan reduces proteinuria in CKD patients. PMID- 21626133 TI - Prediction of prognosis after radical cystectomy for pathologic node-negative bladder cancer. AB - OBJECTIVE: To evaluate risk factors related to bladder cancer recurrence in patients with pathologically negative pelvic lymph nodes. METHODS: We reviewed 192 patients who underwent radical cystectomy for muscle invasive bladder cancer between 1999 and 2009. Patients with pathologically positive pelvic lymph nodes and <6-month follow-up were excluded. Clinicopathological parameters including grade, stage, lymphadenectomy type (standard vs. extended), number of nodes removed, margin status, lymphovascular invasion, perineural invasion, presence of carcinoma in situ, ureteral invasion, prostatic urethral invasion, and longest tumor dimension were evaluated. Kaplan-Meier and multivariate Cox analyses were used to evaluate these parameters for 5-year recurrence-free survival (5RFS) and 5-year overall survival (5OS). RESULTS: One hundred and fifty-five patients were included in this study. Mean follow-up duration was 36.6 months (12-141 months). Nuclear grade according to 2004 WHO classification (P = 0.006, P = 0.001), perineural invasion (P = 0.041, P = 0.048), lymphovascular invasion (P = 0.005, P = 0.027), and number of nodes removed (P < 0.001, P < 0.001) demonstrated differences in 5RFS and 5OS, respectively. In multivariate analysis, 5RFS was related to perineural invasion (P = 0.008; HR: 10.951; 95% CI: 1.891-63.407) and the number of nodes removed (P = 0.038; HR: 1.087; 95% CI: 1.004-1.176), and 5OS was correlated with lymphovascular invasion (P = 0.035; HR: 6.694; 95% CI: 1.143 39.217). CONCLUSIONS: Perineural invasion and the number of nodes removed were independent risk factors for 5RFS, and lymphovascular invasion was for 5OS in patients with node-negative bladder cancer. PMID- 21626134 TI - Leclercia adecarboxylata, an unusual hemodialysis catheter-related infection. PMID- 21626135 TI - Fluoride content and distribution pattern in groundwater of eastern Yunnan and western Guizhou, China. AB - For study, the fluoride (F) content and distribution pattern in groundwater of eastern Yunnan and western Guizhou fluorosis area in southwestern China, the F content of 93 water samples [groundwater (fissure water, cool spring, and hot springs), rivers water] and 60 rock samples were measured. The result shows the F content of the fissure water and cold spring water is 0.027-0.47 mg/L, and river water is 0.048-0.224 mg/L. The F content of hot spring water is 1.02-6.907 mg/L. The drinking water supplied for local resident is mainly from fissure water, cool spring, and river water. And the F content in all of them is much lower than the Chinese National Standard (1.0 mg/L), which is the safe intake of F in drinking water. The infected people in eastern Yunnan and western Guizhou fluorosis area have very little F intake from the drinking water. The hot spring water in fluorosis area of eastern Yunnan and western Guizhou, southwest China has high F content, which is not suitable for drinking. PMID- 21626136 TI - Developmental changes in mesenteric artery reactivity in embryonic and newly hatched chicks. AB - At birth, the intestine becomes the sole site for nutrient absorption requiring a dramatic increase in blood flow. The vascular changes accompanying this transition have been partly characterized in mammals. We investigated, using wire myography, the developmental changes in chick mesenteric artery (MA) reactivity. Rings of the MA from 15-day (E15) and 19-day (E19) chicken embryos (total incubation 21 days) as well as non-fed 0-3-h-old (NH3h) and first-fed 1-day-old (NH1d) newly hatched chicks contracted in response to KCl, norepinephrine (NE), U46619, and endothelin (ET)-1 and relaxed in response to acetylcholine (ACh), sodium nitroprusside (SNP), and forskolin indicating the presence of electro- and pharmaco-mechanical coupling as well as cGMP- and cAMP-mediated relaxation. In ovo development and transition to ex ovo life was accompanied by alterations in the response of the MAs, but a different developmental trajectory was observed for each reactivity pathway tested. Thus, the contractile efficacy of KCl underwent a linear increase (E15 < E19 < NH3h < NH1d). The efficacy of NE and U46619 increased in ovo, but not ex ovo (E15 < E19 = NH3h = NH1d) and the efficacy of ET-1 peaked at E19 (E15 < E19 > NH3h = NH1d). The relaxations elicited by ACh (endothelium-dependent), SNP, and forskolin did not undergo significant developmental changes. In conclusion, the ability of chick MAs to constrict in response to pharmacological stimuli increases during the embryonic period, but no dramatic changes are induced by hatching or the first feeding. Maturation of vasodilator mechanisms precedes that of vasoconstrictor mechanisms. Alterations of the delicate balance between vasoconstrictors and vasodilators may play an important role in perinatal intestinal diseases. PMID- 21626137 TI - Two-marker association tests yield new disease associations for coronary artery disease and hypertension. AB - It has been postulated that multiple-marker methods may have added ability, over single-marker methods, to detect genetic variants associated with disease. The Wellcome Trust Case Control Consortium (WTCCC) provided the first successful large genome-wide association studies (GWAS) which included single-marker association analyses for seven common complex diseases. Of those signals detected, only one was associated with coronary artery disease (CAD), and none were identified for hypertension (HTN). Our objective was to find additional genetic associations and pathways for cardiovascular disease by examining the WTCCC data for variants associated with CAD and HTN using two-marker testing methods. We applied two-marker association testing to the WTCCC dataset, which includes ~2,000 affected individuals with each disorder, and a shared pool of ~3,000 controls, all genotyped using Affymetrix GeneChip 500 K arrays. For CAD, we detected single nucleotide polymorphisms (SNP) pairs in three genes showing genome-wide significance: HFE2, STK32B, and DIPC2. The most notable SNP pairs in a non-protein-coding region were at 9p21, a known major CAD-associated region. For HTN, we detected SNP pairs in five genes: GPR39, XRCC4, MYO6, ZFAT, and MACROD2. Four further associated SNP pair regions were at least 70 kb from any known gene. We have shown that novel, multiple-marker, statistical methods can be of use in finding variants in GWAS. We describe many new, associated variants for both CAD and HTN and describe their known genetic mechanisms. PMID- 21626139 TI - In search of the unknown: introduction to the special issue. AB - The problem of aesthetic perception occupied Vygotsky throughout his life. Working in different research collectives or networks he worked out different answers but never reached a final solution. Inadequate and incomplete access to his writings unfortunately hinders us from understanding Vygotsky's ideas and his personal motives. Publication of his notebooks and unadulterated versions of his writings plus an analysis of his research networks will deepen our understanding. PMID- 21626138 TI - Xist regulation and function explored. AB - X chromosome inactivation (XCI) is a process in mammals that ensures equal transcript levels between males and females by genetic inactivation of one of the two X chromosomes in females. Central to XCI is the long non-coding RNA Xist, which is highly and specifically expressed from the inactive X chromosome. Xist covers the X chromosome in cis and triggers genetic silencing, but its working mechanism remains elusive. Here, we review current knowledge about Xist regulation, structure, function and conservation and speculate on possible mechanisms by which its action is restricted in cis. We also discuss dosage compensation mechanisms other than XCI and how knowledge from invertebrate species may help to provide a better understanding of the mechanisms of mammalian XCI. PMID- 21626140 TI - To Moscow with love: partial reconstruction of Vygotsky's trip to London. AB - The Russian psychologist Lev Vygotsky (1896-1934) left the Soviet Union only once to attend a conference on the education of the deaf in London. So far almost nothing was known about this trip, which took place in a period when Vygotsky was still completely unknown as a psychologist, both inside his own country and abroad. Making use of a newly discovered notebook, it proved possible to partially reconstruct Vygotsky's journey and stay in London. Vygotsky's very personal remarks show him to have been a very sensitive and spirited man, who was prey to strong emotions during the conference and afterwards. Rather surprisingly, Vygotsky's own paper about the education of the deaf was never presented during the conference and the stay in London appears to have had a limited value for his own scientific development. PMID- 21626141 TI - Vygotsky in english: what still needs to be done. AB - At present readers of English have still limited access to Vygotsky's writings. Existing translations are marred by mistakes and outright falsifications. Analyses of Vygotsky's work tend to downplay the collaborative and experimental nature of his research. Several suggestions are made to improve this situation. New translations are certainly needed and new analyses should pay attention to the contextual nature of Vygotsky's thinking and research practice. PMID- 21626142 TI - Hypericin and pseudohypericin concentrations of a valuable medicinal plant Hypericum perforatum L. are enhanced by arbuscular mycorrhizal fungi. AB - Hypericum perforatum L. (St. John's-wort, Hypericaceae) is a valuable medicinal plant species cultivated for pharmaceutical purposes. Although the chemical composition and pharmacological activities of H. perforatum have been well studied, no data are available concerning the influence of arbuscular mycorrhizal fungi (AMF) on this important herb. A laboratory experiment was therefore conducted in order to test three AMF inocula on H. perforatum with a view to show whether AMF could influence plant vitality (biomass and photosynthetic activity) and the production of the most valuable secondary metabolites, namely anthraquinone derivatives (hypericin and pseudohypericin) as well as the prenylated phloroglucinol-hyperforin. The following treatments were prepared: (1) control-sterile soil without AMF inoculation, (2) Rhizophagus intraradices (syn. Glomus intraradices), (3) Funneliformis mosseae (syn. Glomus mosseae), and (4) an AMF Mix which contained: Funneliformis constrictum (syn. Glomus constrictum), Funneliformis geosporum (syn. Glomus geosporum), F. mosseae, and R. intraradices. The application of R. intraradices inoculum resulted in the highest mycorrhizal colonization, whereas the lowest values of mycorrhizal parameters were detected in the AMF Mix. There were no statistically significant differences in H. perforatum shoot mass in any of the treatments. However, we found AMF species specificity in the stimulation of H. perforatum photosynthetic activity and the production of secondary metabolites. Inoculation with the AMF Mix resulted in higher photosynthetic performance index (PI(total)) values in comparison to all the other treatments. The plants inoculated with R. intraradices and the AMF Mix were characterized by a higher concentration of hypericin and pseudohypericin in the shoots. However, no differences in the content of these metabolites were detected after the application of F. mosseae. In the case of hyperforin, no significant differences were found between the control plants and those inoculated with any of the AMF applied. The enhanced content of anthraquinone derivatives and, at the same time, better plant vitality suggest that the improved production of these metabolites was a result of the positive effect of the applied AMF strains on H. perforatum. This could be due to improved mineral nutrition or to AMF-induced changes in the phytohormonal balance. Our results are promising from the biotechnological point of view, i.e. the future inoculation of H. perforatum with AMF in order to improve the quality of medicinal plant raw material obtained from cultivation. PMID- 21626143 TI - Edible sporocarp production by age class in a Scots pine stand in Northern Spain. AB - With the aim of increasing knowledge of community structure, dynamics and production of ectomycorrhizal fungi, edible sporocarp yields were monitored between 1995 and 2004 in a Pinus sylvestris stand in the northeast zone of the Iberian Peninsula. A random sampling design was performed by stand age class according to the forest management plan: 0-15, 16-30, 31-50, 51-70 and over 71 years-old. Eighteen 150 m plots were established and sampled weekly every year from September to December. One hundred and nineteen taxa belonging to 51 genera were collected, 40 of which were edible and represented 74% of the total biomass. Boletus edulis, Lactarius deliciosus, Cantharellus cibarius and Tricholoma portentosum sporocarps, which are considered to be of high commercial value, represented 34% of the total production. B. edulis and L. deliciosus were the most remarkable and abundant species, and both were collected in more than 60% of the samplings. B. edulis fructified every year of the experiment; its mean production was 40 kg/ha and year and its maximum productivity was more than 94 kg/ha in 1998. The age class with the largest production of this taxa was the fourth (51-70 years), with 70 kg/ha. L. deliciosus only failed to fructify one autumn (2000); its mean production was almost 10 kg/ha and its maximum productivity close to 30 kg/ha in 1997. The maximum productivity of this species was found in the second (16-30 years) and fifth (71-90 years) stand age classes, with 18 and 16 kg/ha, respectively. Advances in this field can certainly offer new insights into factors affecting sporocarp production. PMID- 21626144 TI - Up-regulation of bradykinin B2 receptor by Pseudomonas aeruginosa via the NF kappaB pathway. AB - As the first line of host defense, inflammatory responses in response to bacterial infection are initiated by the production of a range of mediators. Infection of Pseudomonas aeruginosa has been shown to stimulate the production of bradykinin (BK), which is known as a universal mediator for the induction of inflammatory reaction via the predominant interaction with the bradykinin B2 receptor (B2R). Thus, the interaction between BK and B2R represents an important host innate response against invading P. aeruginosa. However, the contribution of P. aeruginosa to the up-regulation of B2R expression remains unclear. Here, we report that P. aeruginosa is potent in inducing the expression of B2R at the mRNA and protein levels in a dose- and time-dependent manner. Components produced and secreted from P. aeruginosa could play an essential role in inducing B2R expression, and the secreted components are not under the control of Type III secretion system or quorum sensing. B2R expression in response to P. aeruginosa is mediated by the induction of cellular signaling that leads to the activation of transcription factor NF-kappaB. Thus, this study demonstrates that P. aeruginosa is able to up-regulate the expression of B2R during infection via the NF-kappaB signaling pathway. PMID- 21626145 TI - Analysis of Escherichia coli isolated from patients affected by Crohn's disease. AB - The etiopathogenesis of Crohn's disease (CD) is still controversial: several genetic, immunologic, and environmental factors, including some bacteria, have been implicated. This study has been devised to assess the involvement of Escherichia coli in CD. Seven E. coli strains were isolated from 14 biopsies obtained from ileocolic ulcers of patients affected by inflammatory bowel disease (IBD), including six with ulcerative colitis and eight with CD. Five strains, exclusively isolated from CD patients, were found inside mucosal cells. Different PCR techniques (for chuA, yjaA, TspE4.C2, escV, and bfpB genes) were performed and PFGE was carried out to characterize these bacteria in comparison with other E. coli strains isolated from non-IBD specimens. The correlation of these characters with bacterial invasiveness on intestinal (Caco-2) and phagocytic (U937) cells was assessed. Overall our pilot data suggest that five among eight strains isolated from CD patients belonged to the adherent-invasive E. coli (AIEC) group, and were invasive on Caco-2 cells and resistant to phagocytosis. These findings suggest that these bacteria could be considered target organisms whose elimination could reduce the intestinal inflammatory process and CD progression. PMID- 21626146 TI - Kanagawa-negative, tdh- and trh-positive Vibrio parahaemolyticus isolated from fresh oysters marketed in Fortaleza, Brazil. AB - Between October 2008 and June 2009, 15 samples of 10 live oysters each (Crassostrea rhizophorae) measuring 8.31-10.71 cm were purchased from a restaurant on the seashore of Fortaleza, Brazil. The Vibrio count ranged from 75 (estimated) to 43,500 CFU/g. Fourteen species were identified among the 56 isolated Vibrio strains, with V. parahaemolyticus as the most prevalent. Two of the 17 V. parahaemolyticus strains were urease-positive and tdh- and trh-positive on multiplex PCR, but neither produced beta-hemolysis halos in Wagatsuma agar. Thus, fresh oysters served in natura in Fortaleza, Brazil, were found to contain Vibrio strains known to cause gastroenteritis in humans. PMID- 21626147 TI - Identification of SACE_7040, a member of TetR family related to the morphological differentiation of Saccharopolyspora erythraea. AB - SACE_7040 is presumed to be a member of the TetR family of transcriptional regulators in Saccharopolyspora erythraea, but its biological function is unknown. It was shown that the SACE_7040 gene knockout mutant formed aerial mycelium earlier than its original strain, and this phenotype could be restored by complementation of a single copy of SACE_7040 gene, demonstrating that SACE_7040 is an important regulator of the morphological differentiation of Sac. erythraea. When SACE_7040 gene was disrupted in the bldD mutant, we intriguingly found that the defect in aerial development exhibited by the bldD mutant could be overcome, suggesting a crosstalk between SACE_7040 and BldD in Sac. erythraea morphogenesis. These findings provide novel insights toward the Sac. erythraea developmental biology. PMID- 21626148 TI - Bellis perennis: a useful tool for protein localization studies. AB - Fluorescent fusion proteins together with transient transformation techniques are commonly used to investigate intracellular protein localisation in vivo. Biolistic transfection is reliable, efficient and avoids experimental problems associated with producing and handling fragile protoplasts. Onion epidermis pavement cells are frequently used with this technique, their excellent properties for microscopy resulting from their easy removal from the underlying tissues and large size. They also have advantages over mesophyll cells for fluorescence microscopy, as they are devoid of chloroplasts whose autofluorescence can pose problems. The arrested plastid development is peculiar to epidermal cells, however, and stands in the way of studies on protein targeting to plastids. We have developed a system enabling studies of in vivo protein targeting to organelles including chloroplasts within a photosynthetically active plant cell with excellent optical properties using a transient transformation procedure. We established biolistic transfection in epidermal pavement cells of the lawn daisy (Bellis perennis L., cultivar "Galaxy red") which unusually contain a moderate number of functional chloroplasts. These cells are excellent objects for fluorescence microscopy using current reporters, combining the advantages of the ease of biolistic transfection, the excellent optical properties of a single cell layer and access to chloroplast protein targeting. We demonstrate chloroplast targeting of plastid-localised heme oxygenase, and two further proteins whose localisation was equivocal. We also demonstrate unambiguous targeting to mitochondria, peroxisomes and nuclei. We thus propose that the Bellis system represents a valuable tool for protein localisation studies in living plant cells. PMID- 21626149 TI - Expression, subcellular localization, and cis-regulatory structure of duplicated phytoene synthase genes in melon (Cucumis melo L.). AB - Carotenoids perform many critical functions in plants, animals, and humans. It is therefore important to understand carotenoid biosynthesis and its regulation in plants. Phytoene synthase (PSY) catalyzes the first committed and rate-limiting step in carotenoid biosynthesis. While PSY is present as a single copy gene in Arabidopsis, duplicated PSY genes have been identified in many economically important monocot and dicot crops. CmPSY1 was previously identified from melon (Cucumis melo L.), but was not functionally characterized. We isolated a second PSY gene, CmPSY2, from melon in this work. CmPSY2 possesses a unique intron/exon structure that has not been observed in other plant PSYs. Both CmPSY1 and CmPSY2 are functional in vitro, but exhibit distinct expression patterns in different melon tissues and during fruit development, suggesting differential regulation of the duplicated melon PSY genes. In vitro chloroplast import assays verified the plastidic localization of CmPSY1 and CmPSY2 despite the lack of an obvious plastid target peptide in CmPSY2. Promoter motif analysis of the duplicated melon and tomato PSY genes and the Arabidopsis PSY revealed distinctive cis-regulatory structures of melon PSYs and identified gibberellin-responsive motifs in all PSYs except for SlPSY1, which has not been reported previously. Overall, these data provide new insights into the evolutionary history of plant PSY genes and the regulation of PSY expression by developmental and environmental signals that may involve different regulatory networks. PMID- 21626150 TI - High resolution scanning electron microscopy of plasmodesmata. AB - Symplastic transport occurs between neighbouring plant cells through functionally and structurally dynamic channels called plasmodesmata (PD). Relatively little is known about the composition of PD or the mechanisms that facilitate molecular transport into neighbouring cells. While transmission electron microscopy (TEM) provides 2-dimensional information about the structural components of PD, 3 dimensional information is difficult to extract from ultrathin sections. This study has exploited high-resolution scanning electron microscopy (HRSEM) to reveal the 3-dimensional morphology of PD in the cell walls of algae, ferns and higher plants. Varied patterns of PD were observed in the walls, ranging from uniformly distributed individual PD to discrete clusters. Occasionally the thick walls of the giant alga Chara were fractured, revealing the surface morphology of PD within. External structures such as spokes, spirals and mesh were observed surrounding the PD. Enzymatic digestions of cell wall components indicate that cellulose or pectin either compose or stabilise the extracellular spokes. Occasionally, the PD were fractured open and desmotubule-like structures and other particles were observed in their central regions. Our observations add weight to the argument that Chara PD contain desmotubules and are morphologically similar to higher plant PD. PMID- 21626151 TI - Co-occurrence of obesity and patterns of alcohol use associated with elevated serum hepatic enzymes in US adults. AB - The aim of this cross-sectional study was to present nationally representative findings on the co-occurrence of obesity and specific patterns of alcohol use associated with elevated serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and gamma-glutamyltransferase (GGT) among adults in the United States. We analyzed data from 8,373 adults aged >= 20 years who participated in the 2005-2008 National Health and Nutrition Examination Survey. We produced prevalence ratios by using the co-occurrence of obesity (i.e., body mass index >= 30.0 kg/m(2) or waist circumference >= 102 cm in men and >= 88 cm in women) and specific patterns of alcohol use (i.e., non-drinkers, non-excessive drinkers, and excessive drinkers) as a predictor; elevations in serum ALT, AST, and GGT were used as an outcome variable while adjusting for covariates in multivariate regression models. Approximately 34.7% of adult men and 38.6% of adult women in the United States had co-occurrence of obesity and any alcohol use, including 16.4% of men and 9.8% of women who had co-occurrence of obesity and excessive drinking during 2005-2008. When compared to male non-drinkers without obesity after multivariate adjustment, male excessive drinkers with obesity were 3.08 (95% CI: 1.80-5.28), 2.42 (95% CI: 1.80-3.26), and 3.15 (95% CI: 1.82-5.46) times more likely to exhibit elevated serum ALT, AST, and GGT, respectively. Similarly, when compared to female non-drinkers without obesity, female excessive drinkers with obesity were 2.36 (95% CI: 1.38-4.04), 3.27 (95% CI: 1.85-5.78), and 3.43 (95% CI: 2.19-5.40) times more likely to have elevated serum ALT, AST, and GGT, respectively. The co-occurrence of obesity and excessive drinking may place adults at an increased risk for potential liver injury. Our study findings provide support for evidence-based clinical and population-based interventions that integrate health behavior change among adults who have these co-occurring risk factors. PMID- 21626152 TI - Origin of sexual isolation in Drosophila ananassae due to founder effects. AB - The origin of sexual isolation is the central event in the evolution of biological species and plays a key role in maintaining biological diversity. Three mass culture stocks of D. ananassae originating from different geographic localities showing no isolation with each other were subjected to different degrees of bottlenecks i.e. one pair, five pairs and ten pairs. These drift lines were passed through flush-crash cycle at every generation with same initial number of founders, and maintained for twenty-seven generations and then the pattern of matings was tested among these nine drift lines involving 36 crosses in total. In 23 of 36 crosses, the difference between homogamic and heterogamic matings was significant and isolation indices were significantly more than zero in one direction only providing evidence for asymmetrical sexual isolation. Further, when Bonferroni test for pair-wise analysis was employed, significant differences between homogamic and heterogamic matings were found in 25 crosses. These findings provide evidence for origin of sexual isolation by founder effects in D. ananassae. PMID- 21626153 TI - Effects of artemether, artesunate and dihydroartemisinin administered orally at multiple doses or combination in treatment of mice infected with Schistosoma japonicum. AB - Artemether and artesunate, derivatives of the antimalarial artemisinin, as well as their main metabolite, dihydroartemisinin, all exhibit antischistosomal activities. The purpose of the current study was to compare the effects of artemether, artesunate and dihydroartemisinin administered orally at multiple doses or combination in treatment of mice infected with Schistosoma japonicum. We carried out experiments with mice, infected with 40 cercariae of S. japonicum, and treated with artemether, artesunate and dihydroartemisinin (all at a single dose of 300 mg/kg, and the dose of the mixed three drugs is also 300 mg/kg) at multiple doses or combination therapy on days 6-8 or 34-36 post-infection. Administration with artemether, artesunate or dihydroartemisinin for 3 successive days reduced total worm burdens by 79.5-86% (30.86 +/- 4.98 of mean total worm burden in control), female worm burdens by 79.4-86.7% (11.29 +/- 2.63 of mean female worm burden in control) (all P values <0.01 vs. control), depending on different treatment protocols given on days 6-8 post-infection. However, no differences were seen between each treatment group (all P > 0.05). While the same treatment was given on days 34-36 post-infection, total worm burden reductions of 73.8-75.8% were achieved (29.44 +/- 3.36 of mean total worm burden in control), which were significant when compared with the untreated control group (all P values <0.01). In all different treatment groups, female worm reductions (ranging from 88.7% to 93.1%, while the mean female worm burden in control is 10.33 +/- 1.80) were consistently higher than the total worm reductions, resulting always in significantly lower female worm burdens when compared to the corresponding control (all P values < 0.01). However, there were no significant differences found between each treatment group (all P values >0.05). It is concluded that artemether, artesunate and dihydroartemisinin can be used to control schistosomiasis japonica, as a strategy to prevent S. japonicum infection. Administration with artemether, artesunate and dihydroartemisinin at multiple doses or in combined treatment damages both juvenile and adult S. japonicum, without statistically significant differences among the three drugs at the same dose. PMID- 21626154 TI - In vitro amoebicidal activity of four Peucedanum species on Acanthamoeba castellanii cysts and trophozoites. AB - Amoebic keratitis is difficult to treat without total efficacy in some patients because of cysts, which is less susceptible than trophozoites to the usual treatments. The aim of this study is to evaluate the in vitro amoebicidal activity of the methanolic extracts of Peucedanum caucasicum, Peucedanum palimbioides, Peucedanum chryseum, and Peucedanum longibracteolatum, which are endemic in Turkish flora except P. caucasicum. Extracts were evaluated for their amoebicidal activities using an inverted light microscope. In the presence of methanolic extracts (ranging from 1.0 to 32.0 mg/ml), numbers of the viable Acanthamoeba castellani trophozoites and cysts were determined during the experimental process (72nd hour). All of the extracts showed a time and dose dependent amoebicidal action on the trophozoites and cysts. Among the extracts tested, P. longibracteolatum showed the strongest amoebicidal effect on the trophozoites and cysts. In the case of 32 mg/ml concentration of extract, no viable trophozoites or cysts were determined between 24th and 72nd hour. Similar results were obtained from the extract at 16.0 mg/ml concentration against trophozoites. At this concentration value, number of viable cysts was determined as 10.6 +/- 2.1 in the 24th hour. In the presence of 8.0 mg/ml extract solution, no viable trophozoites were determined in the 48th hour. At the same concentration, 51% of the cysts were killed by the extract in the 72nd hour. As expected, cysts were found more resistant to the extracts than the trophozoites. PMID- 21626155 TI - The secretions products from invading cercariae of S. japonicum (0-3hRP) restrain mouse dendritic cells to mature. AB - Schistosomiasis is a water-borne infection caused mainly by human schistosomes including Schistosoma mansoni (S. mansoni) and Schistosoma japonicum (S. japonicum). In the infected host, immune events in the skin appear to have an important initiating role in determining the type, and guiding the magnitude, of the ensuing acquired immune response. The previous studies of S. mansoni showed that dendritic cells (DCs) prime Th2 response after activating with products released by schistosome larvae (material released in the first 3 h after transformation, 0-3hRP). Therefore, it is interesting to know whether 0-3hRP from S. japonicum also activate DC and induce Th2 immune response. Here, we report 0 3hRP of S. japonicum failed to stimulate the maturation of bone marrow-derived DCs (BM-DCs), while soluble antigen of the body of cercariae preparation (SCAP) induced BM-DCs to mature which can be weakened by 0-3hRP. Moreover, using an in vitro ovalbumin peptide-restricted priming assay, DCs treated with 0-3hRP failed to induce T cells to secrete more cytokines than the negative control group by CD4(+) T cells from DO11.10 transgenic mice, while DCs treated with SCAP upregulated secretions of IFN-gamma and IL-17A, but downregulated IL-4. Importantly, BM-DCs treated with 0-3hRP plus SCAP induced BM-DCs selective mature which drive Th2-type polarized response. Our in vitro results agree with the findings of in vivo studies by inoculation of DO11.10 mice with different stimulus-activated DCs pulsed with ovalbumin peptide. Our data demonstrate that the secretions from invading cercariae of S. japonicum (0-3hRP) impaired DCs to mature, which is potentially allowing the parasite to negotiate the immune recognition and attack. PMID- 21626156 TI - Highly debilitating natural Trypanosoma vivax infections in Brazilian calves: epidemiology, pathology, and probable transplacental transmission. AB - Clinical, epidemiological, and pathological aspects of trypanosomiasis caused by Trypanosoma vivax in calves were reported for the first time in northeast Brazil. Clinical and epidemiological data, packed cell volumes (PCV), and parasitemia were assessed in 150 calves in May 2009 (rainy season-survey 1) and in 153 calves in November 2009 (dry season-survey 2) in three farms (A, B, and C). Prevalence of T. vivax in calves examined in the survey 1 was 63.3%, 65.0%, and 80.0% in farms A, B, and C, respectively. Morbidity varied from 63.3% to 80%, mortality from 15% to 30% and lethality from 23% to 37.5%. In survey 1, for all farms, high parasitemia (from 30.3 to 26.2 * 10(6) parasites/mL), fever (from 39.8 to 40.3 degrees C), low PCV (from 15.7% to 18.1%), and body score (from 2.5 to 3.5) were detected. Calves showed depression, weight loss, pale mucous membranes, enlarged lymph nodes, edema of the dewlap, cough, coryza, and diarrhea. The animals from farms A and B were treated with diminazene aceturate. Six months after, in survey 2, non-treated calves from farm C showed values for prevalence (81.82), morbidity (81.82), mortality (12.73), and lethality (15.55) similar to those in survey 1 (P > 0.05). Also in survey 2, four calves aging merely 1-3 days old presented high parasitemia levels (from 32 * 10(6) to 74 * 10(6) parasites/mL), suggesting transplacental transmission. In conclusion, trypanosomiasis by T. vivax constitutes high prevalent disease for calves raised in Brazilian semiarid and may have transplacental transmission. PMID- 21626157 TI - Resistance differences between chlorpyrifos and synthetic pyrethroids in Cimex lectularius population from Denmark. AB - Bed bug, Cimex lectularius L., populations were investigated for resistance against permethrin and chlorpyrifos in a topical application bioassay, after an initial establishment of a discriminating dose with a susceptible population. For both insecticides, ca. two times the lethal dose LD(99) was selected: 2,560 ng of permethrin and 200 ng of chlorpyrifos per bed bug, respectively. Bed bugs were collected from infested homes in Denmark at ten locations and bred in the laboratory. The frequency of permethrin-resistant individuals was high in Danish bed bug populations as susceptible individuals were only found in three of ten populations. In contrast, the frequency of chlorpyrifos-resistant individuals was low in Danish bed bug populations, but resistant individuals were found in five of ten populations. To test the significance of the observed resistance, we performed tarsal contact test with commercially available insecticides. The test indicated that both a permethrin and a deltamethrin product had very low efficacy against the field-collected bed bug populations. Despite the reduced sensitivity to synthetic pyrethroids, all populations tested in the tarsal test on the commercial product with micro-encapsulated chlorpyrifos resulted in close to 100% mortality. PMID- 21626158 TI - Structural insights on two hypothetical secretion chaperones from Xanthomonas axonopodis pv. citri. AB - Several Gram-negative bacterial pathogens have developed type III secretion systems (T3SSs) to deliver virulence proteins directly into eukaryotic cells in a process essential for many diseases. The type III secretion processes require customized chaperones with high specificity for binding partners, thus providing the secretion to occur. Due to the very low sequence similarities among secretion chaperones, annotation and discrimination of a great majority of them is extremely difficult and a task with low scores even if genes are encountered that codify for small (<20 kDa) proteins with low pI and a tendency to dimerise. Concerning about this, herein, we present structural features on two hypothetical T3SSs chaperones belonging to plant pathogen Xanthomonas axonopodis pv. citri and suggest how low resolution models based on Small Angle X-ray Scattering patterns can provide new structural insights that could be very helpful in their analysis and posterior classification. PMID- 21626159 TI - Thermal denaturation of beta-glucosidase B from Paenibacillus polymyxa proceeds through a Lumry-Eyring mechanism. AB - beta-glucosidase B (BglB), 1,4-beta-D: -glucanohydrolase, is an enzyme with various technological applications for which some thermostable mutants have been obtained. Because BglB denatures irreversibly with heating, the stabilities of these mutants are assessed kinetically. It, therefore, becomes relevant to determine whether the measured rate constants reflect one or several elementary kinetic steps. We have analyzed the kinetics of heat denaturation of BglB from Paenibacillus polymyxa under various conditions by following the loss of secondary structure and enzymatic activity. The denaturation is accompanied by aggregation and an initial reversible step at low temperatures. At T >= T ( m ), the process follows a two-state irreversible mechanism for which the kinetics does not depend on the enzyme concentration. This behavior can be explained by a Lumry-Eyring model in which the difference between the rates of the irreversible and the renaturation steps increases with temperature. Accordingly, at high scan rates (>=1 degrees C min(-1)) or temperatures (T >= T ( m )), the measurable activation energy involves only the elementary step of denaturation. PMID- 21626160 TI - Teriparatide in bisphosphonate-resistant osteoporosis: microarchitectural changes and clinical results after 6 and 18 months. AB - A number of osteoporotic patients under bisphosphonate treatment present persistent fragility fractures and bone loss despite good compliance. The objective of this 18-month prospective study was to investigate the effect of teriparatide [rhPTH(1-34)] in 25 female osteoporotics who were inadequate responders to oral bisphosphonates and to correlate microarchitectural changes in three consecutive iliac crest biopsies measured by micro-computed tomography (MUCT) with bone mineral density (BMD) and bone serum markers. Scanned biopsies at baseline (M0), 6 months (M6), and 18 months (M18) demonstrated early significant (P < 0.01) increases in bone volume per tissue volume (+34%) and trabecular number (+14%) at M6 with only moderate changes in most MUCT structural parameters between M6 and M18. MUCT-measured bone tissue density was significantly decreased at M18, expressing an overall lower degree of tissue mineralization characteristic for new bone formation despite unchanged trabecular thickness due to increased intratrabecular tunneling at M18. MUCT results were consistent with serum bone turnover markers, reaching maximal levels of bone alkaline phosphatase and serum beta-crosslaps at M6, with subsequent decline until M18. BMD assessed by DXA demonstrated persistent increases at the lumbar spine until M12, whereas no significant change was observed at the hip. Type (alendronate/risedronate) and duration (3.5 +/- 4 years) of prior bisphosphonate treatment did not influence outcome on MUCT, BMD, or bone marker results. The overall results indicate a positive ceiling effect of teriparatide on bone microarchitecture and bone markers after 6 and 12 months for lumbar spine BMD, with no additional gain until M18 in bisphosphonate nonresponders. PMID- 21626161 TI - Introduction: Immunopathology and the promise of stem cell research. PMID- 21626162 TI - Prognostic impact of anticancer immune responses: an introduction. AB - Cancer cells escape innate and adaptive immune responses by selection of non immunogenic tumor cell variants (immunoediting) or by active suppression of the immune response (immunosubversion). One common strategy that employs tumor cells to elude a T-cell-mediated immune response is the downregulation or loss of expression of HLA class I molecules, often associated with an induction of the surface expression of HLA class II antigen, nonclassical HLA class I molecules, and/or NK cell-activating ligands such as MICA, MICB, or ULBP. These changes in the surface characteristics of tumor cells result from the selection pressure exerted by immune cells from the innate and adaptive compartment. The tumor antigen characteristics are "immunoedited" in the course of the disease, resulting in the survival of tumor variants with defective antigen presentation, mostly at the level of HLA class I. PMID- 21626164 TI - Xanthogranulomatous variant of immunoglobulin G4 sclerosing disease presenting as ptosis, proptosis and eyelid skin plaques. AB - A 70-year-old male was referred to the oculoplastic clinic with left-sided ptosis and floppy eyelids. During follow-up, bilateral upper lid xanthelasma developed with worsening ptosis and proptosis, which was worse on the left side. A left orbital biopsy showed xanthogranulomatous inflammation of the orbit. The patient was treated with a variety of immune modulator regimes but due to a variety of side-effects, treatment was discontinued. The left orbit was surgically debulked twice and histology revealed xanthogranulomatous inflammation, with the additional features of sclerosis, lymphoid aggregates and a prominent population of plasma cells. Around 80% of the plasma cells expressed immunoglobulin G4 (IgG4). This case report reveals an association between xanthogranulomatous inflammation of the orbit and a prominent population of IgG4-positive plasma cells. We propose that the overall disease is a novel variant of IgG4 sclerosing disease of the orbit and suggest that cases of histologically proven xanthogranulomatous inflammation should be stained for IgG4 if there is an accompanying plasma cell population. PMID- 21626165 TI - New ophthalmic features in a family with triple A syndrome. AB - We report three subjects of a Greek family affected by triple A syndrome (AAAS). All patients underwent complete ophthalmic examination, full-field electroretinogram (ERG), visual evoked responses (VER), optical coherence tomography (OCT) and molecular analysis of the AAA gene. All patients had alacrima. In two of them, the proband and her brother, bilateral optic atrophy was assessed and the VER were pathological. In contrast, the ERG was normal. OCT showed a decrease of the retinal nerve fiber layer. The third case had only alacrima and the optic nerves were normal. The molecular genetic study of the AAAS gene revealed a homozygous missense mutation p.Ala167Val. To our knowledge this is the first time a family with AAAS has been investigated using OCT, VER and ERG. Our findings illustrate that the retina is not involved. There is also an interfamilial variability concerning the involvement of the optic nerves. PMID- 21626163 TI - Diabetic cardiomyopathy: understanding the molecular and cellular basis to progress in diagnosis and treatment. AB - Diabetes mellitus is an important and prevalent risk factor for congestive heart failure. Diabetic cardiomyopathy has been defined as ventricular dysfunction that occurs in diabetic patients independent of a recognized cause such as coronary artery disease or hypertension. The disease course consists of a hidden subclinical period, during which cellular structural insults and abnormalities lead initially to diastolic dysfunction, later to systolic dysfunction, and eventually to heart failure. Left ventricular hypertrophy, metabolic abnormalities, extracellular matrix changes, small vessel disease, cardiac autonomic neuropathy, insulin resistance, oxidative stress, and apoptosis are the most important contributors to diabetic cardiomyopathy onset and progression. Hyperglycemia is a major etiological factor in the development of diabetic cardiomyopathy. It increases the levels of free fatty acids and growth factors and causes abnormalities in substrate supply and utilization, calcium homeostasis, and lipid metabolism. Furthermore, it promotes excessive production and release of reactive oxygen species, which induces oxidative stress leading to abnormal gene expression, faulty signal transduction, and cardiomyocytes apoptosis. Stimulation of connective tissue growth factor, fibrosis, and the formation of advanced glycation end-products increase the stiffness of the diabetic hearts. Despite all the current information on diabetic cardiomyopathy, translational research is still scarce due to limited human myocardial tissue and most of our knowledge is extrapolated from animals. This paper aims to elucidate some of the molecular and cellular pathophysiologic mechanisms, structural changes, and therapeutic strategies that may help struggle against diabetic cardiomyopathy. PMID- 21626166 TI - Triamcinolone acetonide-assisted vitrectomy for stage 4 retinopathy of prematurity. PMID- 21626168 TI - Herpes zoster optic neuritis. AB - Herpes zoster (HZ) is an acute infection caused by reactivation of the latent varicella-zoster virus [1]. Herpes zoster ophthalmicus (HZO) occurs when inflammation spreads from the ganglion of Gasser to the ophthalmic branch of the trigeminal nerve. Optic neuritis, a very rare sequela of HZO [2-4], can occur simultaneously to the acute vesicular skin eruption or, more frequently, as a postherpetic complication. We report on a 74-year-old woman who presented with HZ optic neuritis 45 days after developing an incompletely treated bout of trigeminal HZ, characterized only by pruritus. It is important to value the non specific manifestations of cutaneous HZ in the prodromal phase, so as to offer timely and appropriate treatment. PMID- 21626167 TI - Classical familial homocystinuria in an adult presenting as an isolated lens subluxation. AB - To report a case of late diagnosis of a classical familial homocystinuria based on an ophthalmologic examination. A 35-year-old male with Marfan-like phenotype complained of a progressive increase of myopia during the previous 2 years. Ophthalmologic exploration showed a bilateral subluxation of the lens with inferior and nasal displacement. Biochemical study detected a profile of increased amino acid levels (homocysteinemia) consistent with suspected homocystinuria. Vascular and skeletal studies ruled out Marfan syndrome. Response to treatment demonstrated B(6)-non-responsive homocystinuria. Molecular study showed compound heterozygous T353 N and D444 N mutations of the cystathionine beta-synthase (CBS) gene, and also a C667T homozygous mutation of the methylenetetrahydrofolate-reductase (MTHFR) gene. Family study showed classical homocystinuria in his father and sister, although they did not present any systemic or ocular features of the disease. Homocystinuria is a metabolic disease usually presenting at an early age as vascular, skeletal and neuropsychiatric abnormalities, as well as ectopia lentis. Our case is atypical because of the absence of thromboembolism and the mild phenotype, in spite of being B(6)-non responsive, and the association of a rare compound heterozygous mutation of the CBS gene and also an homozygous mutation of the MTHFR gene. It is necessary to rule out homocystinuria in patients with ectopia lentis, even in the absence of systemic symptoms. PMID- 21626169 TI - Neurochemical plasticity of nitric oxide synthase isoforms in neurogenic detrusor overactivity after spinal cord injury. AB - Nitric oxide (NO) participates in the neural pathways controlling the lower urinary tract (LUT). Expression of NO synthase (NOS) can be upregulated after spinal cord injury (SCI), and altered NOS activity may participate in resulting LUT dysfunction. To investigate distribution of NOS-immunoreactivity (NOS-IR) in neurons of rats following SCI and the possible effects of NOS inhibitors. Expression of neuronal and inducible NOS-IR in lumbosacral spinal cord was assessed in rats. Cystometry was performed to examine effects of intrathecal injection of NOS inhibitor. There was increased expression of neuronal NOS-IR after trauma. Maximum bladder capacity was increased by neuronal NOS (nNOS) inhibitors. Upregulation of nNOS may facilitate emergence of the spinal micturition reflex following SCI; nNOS inhibitor suppressed SCI-induced urinary incontinence by increasing bladder capacity. Our results indicate manipulation of NO production could help treat LUT dysfunction after SCI. PMID- 21626170 TI - Castration attenuates myelin repair following lysolecithin induced demyelination in rat optic chiasm: an evaluation using visual evoked potential, marker genes expression and myelin staining. AB - Multiple sclerosis (MS) is a demyelinating disease that affects the central nervous system. MS is the most common neurological disorder in young adults with a greater incidence among females. Male gonadal hormones have a protective effect on neural system development and myelin maturation. In this study, we investigate the effect of castration on lysolecithin-induced demyelination and remyelination processes using visual evoked potentials, in addition to measuring the expressions of Olig2, MBP, Nogo-A and GFAP mRNAs as oligodendrocyte or astrocyte markers; and histological assessments by myelin-specific staining. We observed more expanded demyelination with delayed repair process in castrated rats. Expression levels of the aforementioned marker genes confirmed histological and electrophysiological observations. Our results showed a pivotal role for endogenous male hormones in the context of demyelinating insults. It may also account for the different prognosis of MS between male and female genders and provide new insights for therapeutic treatments. PMID- 21626171 TI - Rare presentation of juvenile xanthogranuloma in the thoracic spine of an adult patient: case report and literature review. AB - Isolated spinal involvement of juvenile xanthogranuloma (JXG) is extremely rare. There are only seven prior published cases of spinal JXG, of which only one has been reported in an adult. We report here the eighth case of spinal JXG and the second in an adult. The patient, a 22-year-old female, presented with progressive upper backache. Radiological examination revealed a well-defined osteolytic hypointense mass in the T7 vertebral body, with a large soft tissue paraspinal extension causing cord compression. Complete resection of the mass was performed, with resolution of symptoms. Histology showed a histiocytic tumour with numerous Touton, foreign body and osteoclastic giant cells, immunopositive for CD68 and vimentin and negative for S100 and CD1a, corresponding to a diagnosis of JXG. Literature regarding spinal JXG is reviewed and discussed. PMID- 21626172 TI - The possible role of neuropeptide Y after spontaneous subarachnoid hemorrhage. AB - OBJECTIVE: Neuropeptide Y (NPY), a highly potent vasoconstrictive neuropeptide, is widely expressed in the human brain, regulating vessel diameter and cerebral blood flow. Earlier studies focusing on the possible role of NPY in the context of aneurismal subarachnoid hemorrhage (SAH) and vasospasm have produced conflicting results. However, despite extensive research efforts, the pathophysiological mechanisms underlying the SAH-related vasospasm and delayed cerebral ischemia (DCI) have not been clarified. We, therefore, attempted to investigate the role of NPY in SAH-induced vasospasm in a larger, well documented patient population utilizing modern analytical tools. We focused on the release of the potent vasoconstrictor NPY in cerebrospinal fluid (CSF) and blood, and its correlation to vasospasm and stroke in the early clinical stage. METHODS: Thirty seven patients with SAH and a control group consisting of 29 patients were included. Eighteen patients developed stroke, 21 patients met the Doppler sonographical criteria for vasospasm. Twenty-nine patients had aneurysms of the anterior circulation and four patients of the posterior circulation. All patients had ventricular drainage inserted and an arterial catheter. Blood and CSF were drawn daily for NPY analysis during a 10-day interval. RESULTS: The levels of NPY in CSF and plasma were significantly higher after SAH than in the control group (p = 0.001). The vasospasm group showed NPY levels in CSF which continuously ranged above the NPY levels of the non-vasospasm group (p = 0.001). Patients with stroke caused by vasospasm had significantly higher levels of NPY (p = 0.001). DISCUSSION: NPY is released excessively into blood and CSF following SAH. Patients with cerebral infarction caused by vasospasm had significantly higher levels of NPY. Our results indicate a certain role for NPY in the pathophysiology of vasospasm due to SAH and justify further studies in this area of research. PMID- 21626173 TI - Impact of postpartum depressive and anxiety symptoms on mothers' emotional tie to their infants 2-3 months postpartum: a population-based study from rural Bangladesh. AB - The purpose of this study was to investigate the impact of depressive and anxiety symptoms on maternal bonding to the infant 2-3 months postpartum and the influence of the mother's bonding to the infant during pregnancy and to her own caregiver during her childhood on maternal bonding 2-3 months postpartum. This study originated from a community-based cohort study carried out in rural Bangladesh. Trained staff collected data and administrated the questionnaires during the third trimester of pregnancy, at childbirth and 2-3 months postpartum. Maternal depressive and anxiety symptoms were assessed with the Edinburgh Postnatal Depression Scale and the State Anxiety Inventory and the mother's emotional bonding to the infant with the Postpartum Bonding Questionnaire. The results showed that 11% of the women reported depressive symptoms, 35% anxiety symptoms, 3.4% both depressive and anxiety symptoms and 51% neither depressive nor anxiety symptoms. Mothers with depressive symptoms were older, were poorer, fewer were literate, reported more intimate partner violence and showed lower emotional bonding to their infants 2-3 months postpartum compared to mentally well and anxious mothers. Approximately 11% of the mothers reported mild bonding disturbances and nearly one third of them showed depressive symptoms. Depressive symptoms and giving birth to a girl were negatively associated to a mother's emotional bonding to her infant, while maternal anxiety symptoms and high bonding to the foetus during pregnancy were positively associated to the mother's emotional bonding to the infant 2-3 months postpartum. PMID- 21626174 TI - Genetic and functional evaluation of MITF as a candidate gene for cutaneous melanoma predisposition in pigs. AB - Cutaneous melanoma arises from transformed melanocytes and is caused mainly by environmental effects such as ultraviolet radiation and to a lesser extent by predisposing genetic variants. Only a few susceptibility genes for cutaneous melanoma have been identified so far in human; therefore, animal models represent a valuable alternative for genetic studies of this disease. In a previous quantitative trait locus (QTL) study, several susceptibility regions were identified in a swine biomedical model, the MeLiM (Melanoblastoma-bearing Libechov minipig) pigs. This article details the fine-mapping of a QTL located on SSC13 (Sus scrofa chromosome 13) through an increase in marker density. New microsatellites were used to confirm the results of the first analysis, and MITF (microphthalmia-associated transcription factor) was selected as a candidate gene for melanoma development. A single-marker association analysis was performed with single-nucleotide polymorphisms (SNPs) spread over the locus, but it did not reveal a significant association with diverse melanoma-related traits. In parallel, MITF alternative transcripts were characterized and their expression was investigated in different porcine tissues. The obtained results showed a complex transcriptional regulation concordant with the one present in other mammals. Notably, the ratio between MITF+ and MITF- isoforms in melanoma samples followed the same pattern as in human tumors, which highlights the adequacy of the MeLiM pig as a model for human melanoma. In conclusion, although MITF does not seem to be the causal gene of the QTL initially observed, we do not exclude a prominent role of its transcription and function in the outbreak and evolution of the tumors observed in pigs. PMID- 21626175 TI - Coexistence of NtCENH3 and two retrotransposons in tobacco centromeres. AB - Although a centromeric DNA fragment of tobacco (Nicotiana tabacum), Nt2-7, has been reported, the overall structure of the centromeres remains unknown. To characterize the centromeric DNA sequences, we conducted a chromatin immunoprecipitation assay using anti-NtCENH3 antibody and chromatins isolated from two ancestral diploid species (Nicotiana sylvestris and Nicotiana tomentosiformis) of N. tabacum and isolated a 178-pb fragment, Nto1 from N. tomentosiformis, as a novel centromeric DNA. Fluorescence in situ hybridization (FISH) showed that Nto1 localizes on 24 out of 48 chromosomes in some cells of a BY-2 cell line. To identify the origins of the Nt2-7 and Nto1, a tobacco bacterial artificial chromosome (BAC) library was constructed from N. tabacum, and then screened by polymerase chain reaction (PCR) with primer sets designed from the Nt2-7 and Not1 DNA sequences. Twelve BAC clones were found to localize on the centromeric regions by FISH. We selected three BAC clones for sequencing and identified two centromeric retrotransposons, NtCR and NtoCR, the DNA sequences of which are similar to that of Nt2-7 and Nto1, respectively. Quantitative PCR analysis using coprecipitated DNA with anti-NtCENH3 clearly showed coexistence of NtCENH3 with both retrotransposons. These results indicate the possibility that these two retrotransposons act as centromeric DNA sequences in tobacco. NtoCR was found to be specific to N. tomentosiformis and T genome of N. tabacum, and a NtCR-like centromeric retrotransposon (TGRIV) exists in tomato. This specificity suggests that the times of amplification of these centromeric retrotransposons were different. PMID- 21626178 TI - [Contrast-enhanced ultrasound and new techniques]. PMID- 21626179 TI - [Contrast-enhanced ultrasound in animal models]. AB - In the past the detection of tumor perfusion was achieved solely via invasive procedures, such as intravital microscopy or with the help of costly modalities, such as multidetector computed tomography (MDCT), magnetic resonance tomography (MRT) or the combined use of positron emission tomography and computed tomography (PET/CT). Ultrasound offers the non-invasive display of organs without usage of ionizing radiation and it is widely available. However, colour-coded ultrasound and power Doppler do not allow the detection of tumor microcirculation. The introduction of contrast-enhanced ultrasound (CEUS) as well as new high-frequency ultrasound probes made it possible to detect and quantify tumor microcirculation with high resolution. CEUS has been used clinically on human beings for more than 10 years. During the last years different tumor models in experimental animals were used for the establishment of this new technique, e.g. in rats, hamsters and mice. CEUS allows the detection of functional parameters, such as the angiogenetic metabolic status of tissue pretreatment and posttreatment. Further research is required to solve the problems of absolute quantification of these perfusion parameters to allow the comparison of CEUS with other modalities (e.g. MRT and CT). PMID- 21626180 TI - 36-year-old man with sudden severe headache. PMID- 21626181 TI - Advance in the treatment of aneurysmal bone cyst of the sacrum. AB - OBJECTIVE: The objective of this study was the review of 11 patients with two different treatment methods used historically for aneurysmal bone cyst in the sacrum. The outcome of both procedures is reported. In addition, the treatment technique of CT-guided percutaneous injections of demineralized bone matrix mixed with bone marrow concentrate is described. MATERIALS AND METHODS: From 1997 to 2008, 11 patients with sacral aneurysmal bone cyst were treated at the Rizzoli Institute, Bologna, Italy. The first seven patients had surgical curettage without bone grafting, chemical adjuvants, or arterial embolization. The last four patients had arterial embolization. The last patient did not respond to arterial embolization and was treated by CT-guided injection of demineralized bone matrix mixed with bone marrow concentrate. RESULTS: Curettage was successful in five out of seven patients. Two patients suffered complications, and two cases had recurrence. Arterial embolization was successful in three of four patients. The patient treated with injection had a good clinical and radiographic result. CONCLUSION: Both surgical and arterial embolization are effective for aneurysmal bone cyst in the sacrum. However, these treatments may lead to complications and recurrence. The use of CT-guided injections of demineralized bone matrix mixed with bone marrow concentrate may be a safe and effective alternative for treatment of these destructive and problematic lesions. PMID- 21626182 TI - Origin of life prize. PMID- 21626183 TI - Hexaplex PCR assay and liquid bead array for detection of stacked genetically modified cotton event 281-24-236*3006-210-23. AB - A hexaplex system based on multiplex polymerase chain reaction (PCR) coupled with liquid bead array was developed to assist detection of stacked genetically modified (GM) cotton event 281-24-236 * 3006-210-23 (Widestrike) expressing two kinds of endotoxin from Bacillus thuringiensis (Bt). The efficiency of this multiplex detection system was assessed. Specific primer sets for simultaneous detection of six targets in the stacked GM cotton event were constructed and used for the PCR assay. Each of the six targets was amplified, and the amplicons could be separated as discrete bands by agarose gel electrophoresis. A liquid bead array assay for the stacked GM cotton was performed using the hexaplex PCR products followed by hybridization between the biotinylated targets and anti tagged microsphere beads. The hybridization products produced fluorescent signals that were detected by the Luminex system. Signal strengths were analyzed by their median fluorescent intensity values. Comparison of the assays showed that results from the liquid bead array using specific probes agreed with those from the PCR, and detection of the different target elements was found to be very specific with no cross-reaction. Therefore, the combination of hexaplex PCR and liquid bead array for detection of stacked GM events can be a useful and efficient system for screening and analyzing multiple transgenes for simultaneous qualitative analysis. PMID- 21626184 TI - Liquid chromatography time-of-flight mass spectrometry following sorptive microextraction for the determination of fungicide residues in wine. AB - This work evaluates the suitability of sorptive microextraction, using disposable silicone sorbents, and liquid chromatography time-of-flight mass spectrometry (LC TOF-MS) for the determination of 15 fungicides in wine. Under optimized conditions, wine samples (10 mL) were diluted with the same volume of ultrapure water and poured in a glass vessel containing a magnetic stirrer and 4 g of sodium chloride. Extractions were performed at room temperature for 4 h, using an inexpensive silicone disk (12 MUL volume) exposed directly to the sample. Thereafter, analytes were recovered with 0.2 mL of acetonitrile. The electrospray ionization (ESI) source was operated in the fast polarity switching mode obtaining, in the same injection, selective LC-MS records (extracted with a mass window of 10 ppm) of compounds rendering [M + H](+) and [M-H](-) ions. The method provided limits of quantification (LOQs) between 0.1 and 2.2 ng mL(-1), linear response ranges up to 500 ng mL(-1), relative recoveries from 75% to 117% and an inter-day variability below 15% for all analytes in red and white wine samples. The feasibility of in situ sample enrichment followed by delayed desorption and analysis is also assessed. PMID- 21626185 TI - Detecting small lung tumors in mouse models by refractive-index microradiology. AB - Refractive-index (phase-contrast) radiology was able to detect lung tumors less than 1 mm in live mice. Significant micromorphology differences were observed in the microradiographs between normal, inflamed, and lung cancer tissues. This was made possible by the high phase contrast and by the fast image taking that reduces the motion blur. The detection of cancer and inflammation areas by phase contrast microradiology and microtomography was validated by bioluminescence and histopathological analysis. The smallest tumor detected is less than 1 mm(3) with accuracy better than 1 * 10(-3) mm(3). This level of performance is currently suitable for animal studies, while further developments are required for clinical application. PMID- 21626186 TI - Anti-doping analysis. PMID- 21626187 TI - Quantification of carcinogenic 4- to 6-ring polycyclic aromatic hydrocarbons in human urine by solid-phase microextraction gas chromatography-isotope dilution mass spectrometry. AB - Polycyclic aromatic hydrocarbons (PAHs) are pollutants found in living and working environments. The aim of this study was to develop a solid-phase microextraction (SPME) gas chromatography (GC)-isotope dilution mass spectrometry method for the quantification of 10 four- to six-ring PAHs in urine samples. Seven of the selected PAHs have been classified as carcinogenic. Under the final conditions, analytes were sampled with a 100-MUm polydimethylsiloxane SPME fibre for 60 min at 80 degrees C and desorbed in the injection port of the GC at 270 degrees C. Fluoranthene, pyrene, benz[a]anthracene, chrysene, benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[a]pyrene, dibenzo[a,h]anthracene, indeno[1,2,3-cd]pyrene and benzo[ghi]perylene were separated using a highly arylene-modified phase capillary column and quantified by MS using eight deuterated PAHs as surrogate internal standards. Limits of quantification (LOQ) were in the 0.5- to 2.2-ng/L range. Validation showed linear dynamic ranges up to 340 ng/L, inter- and intra-run precisions <20%, and accuracies within 20% of spiked concentrations. Matrix effect evaluation and the use of control charts to monitor process performances showed that the isotope dilution approach allowed for the control of bias sources. Urinary PAHs were above or equal to LOQ, depending on different compounds, in 58-100% (min-max), 40 100% and 5-39% of samples from coke oven workers (n = 12), asphalt workers (n = 10) and individuals not occupationally exposed to PAHs (n = 18), respectively. Chrysene was the most abundant PAH determined with median levels of 62.6, 6.9 and <0.6 ng/L, respectively. These results show that the method is suitable for quantifying carcinogenic PAHs in specimens from individuals with different levels of PAH exposure. PMID- 21626188 TI - Micro-algal biosensors. AB - Fighting against water pollution requires the ability to detect pollutants for example herbicides or heavy metals. Micro-algae that live in marine and fresh water offer a versatile solution for the construction of novel biosensors. These photosynthetic microorganisms are very sensitive to changes in their environment, enabling the detection of traces of pollutants. Three groups of micro-algae are described in this paper: chlorophyta, cyanobacteria, and diatoms. PMID- 21626189 TI - Absolute protein quantification by LC-ICP-MS using MeCAT peptide labeling. AB - Nowadays, the most common strategies used in quantitative proteomics are based on isotope-coded labeling followed by specific molecule mass spectrometry. The implementation of inductively coupled plasma mass spectrometry (ICP-MS) for quantitative purposes can solve important drawbacks such as lack of sensitivity, structure-dependent responses, or difficulties in absolute quantification. Recently, lanthanide-containing labels as metal-coded affinity tag (MeCAT) reagents have been introduced, increasing the interest and scope of elemental mass spectrometry techniques for quantitative proteomics. In this work one of the first methodologies for absolute quantification of peptides and proteins using MeCAT labeling is presented. Liquid chromatography (LC) interfaced to ICP-MS has been used to separate and quantify labeled peptides while LC coupled to electrospray ionization mass spectrometry served for identification tasks. Synthetic-labeled peptides were used as standards to calibrate the response of the detector with compounds as close as possible to the target species. External calibration was employed as a quantification technique. The first step to apply this approach was MeCAT-Eu labeling and quantification by isotope dilution ICP-MS of the selected peptides. The standards were mixed in different concentrations and subjected to reverse-phase chromatography before ICP-MS detection to consider the column effect over the peptides. Thus, the prepared multi-peptide mix allowed a calibration curve to be obtained in a single chromatographic run, correcting possible non-quantitative elutions of the peptides from the column. The quantification strategy was successfully applied to other labeled peptides and to standard proteins such as digested lysozyme and bovine serum albumin. PMID- 21626190 TI - Quality assurance and quality control for thermal/optical analysis of aerosol samples for organic and elemental carbon. AB - Accurate, precise, and valid organic and elemental carbon (OC and EC, respectively) measurements require more effort than the routine analysis of ambient aerosol and source samples. This paper documents the quality assurance (QA) and quality control (QC) procedures that should be implemented to ensure consistency of OC and EC measurements. Prior to field sampling, the appropriate filter substrate must be selected and tested for sampling effectiveness. Unexposed filters are pre-fired to remove contaminants and acceptance tested. After sampling, filters must be stored in the laboratory in clean, labeled containers under refrigeration (<4 degrees C) to minimize loss of semi-volatile OC. QA activities include participation in laboratory accreditation programs, external system audits, and interlaboratory comparisons. For thermal/optical carbon analyses, periodic QC tests include calibration of the flame ionization detector with different types of carbon standards, thermogram inspection, replicate analyses, quantification of trace oxygen concentrations (<100 ppmv) in the helium atmosphere, and calibration of the sample temperature sensor. These established QA/QC procedures are applicable to aerosol sampling and analysis for carbon and other chemical components. PMID- 21626191 TI - Anti-oxidative and inflammatory responses induced by fly ash particles and carbon black in lung epithelial cells. AB - Combustion-derived nanoparticles as constituents of ambient particulate matter have been shown to induce adverse health effects due to inhalation. However, the components inducing these effects as well as the biological mechanisms are still not fully understood. The fine fraction of fly ash particles collected from the electrostatic precipitator of a municipal solid waste incinerator was taken as an example for real particles with complex composition released into the atmosphere to study the mechanism of early biological responses of BEAS-2B human lung epithelial cells. The studies include the effects of the water-soluble and insoluble fractions of the fly ash and the well-studied carbon black nanoparticles were used as a reference. Fly ash induced reactive oxygen species (ROS) and increased the total cellular glutathione (tGSH) content. Carbon black also induced ROS generation; however, in contrast to the fly ash, it decreased the intracellular tGSH. The fly ash-induced oxidative stress was correlated with induction of the anti-oxidant enzyme heme oxygenase-1 and increase of the redox sensitive transcription factor Nrf2. Carbon black was not able to induce HO-1. ROS generation, tGSH increase and HO-1 induction were only induced by the insoluble fraction of the fly ash, not by the water-soluble fraction. ROS generation and HO-1 induction were markedly inhibited by pre-incubation of the cells with the anti-oxidant N-acetyl cysteine which confirmed the involvement of oxidative stress. Both effects were also reduced by the metal chelator deferoxamine indicating a contribution of bioavailable transition metals. In summary, both fly ash and carbon black induce ROS but only fly ash induced an increase of intracellular tGSH and HO-1 production. Bioavailable transition metals in the solid water-insoluble matrix of the fly ash mostly contribute to the effects. PMID- 21626192 TI - Modulation in comprehensive two-dimensional gas chromatography: 20 years of innovation. AB - With almost 20 years having passed since John B. Phillips described the first comprehensive two-dimensional gas chromatography (GC * GC) separation, much has occurred in this ever-expanding field of separation science. GC * GC is currently one of the most effective techniques for the separation and analysis of complex mixtures, offering significantly greater peak capacities than conventional chromatographic methods. The technique is generally based upon separations performed on two chromatographic columns characterized by considerably different selectivities, joined together through a modulating interface. The modulator periodically traps or samples the primary column effluent, usually refocuses it into a narrow chromatographic band and injects the focused fraction into the secondary column. The modulator is often referred to as the 'heart' of the instrument, since a GC * GC separation is impossible without its use. This article reviews major innovations in GC * GC modulator development since its first use by Phillips in 1991. Emphasis has been placed on modulator design and function. PMID- 21626193 TI - GC/MS-based metabolomic approach to validate the role of urinary sarcosine and target biomarkers for human prostate cancer by microwave-assisted derivatization. AB - A recent study showed that sarcosine may be potentially useful for the diagnosis and prognosis of prostate cancer (PCa). The aim of this study was to validate diagnostic value of sarcosine for PCa, to evaluate urine metabolomic profiles in patients with PCa in comparison of non-cancerous control, and to further explore the other potential metabolic biomarkers for PCa. Isotope dilution gas chromatography/mass spectrometry (ID GC/MS) metabolomic approach was applied to evaluate sarcosine using [methyl-D(3)]-sarcosine as an internal standard. Microwave-assisted derivatization (MAD) together with GC/MS was utilized to obtain the urinary metabolomic information in 20 PCa patients compared with eight patients with benign prostate hypertrophy and 20 healthy men. Acquired metabolomic data were analyzed using a two-sample t test. Diagnostic models for PCa were constructed using principal component analysis and were assessed with receiver-operating characteristic curves. Results showed that the urinary sarcosine level has no statistical difference between the PCa group and the control group. In addition, nine metabolomic markers between the PCa group and the healthy male group were selected, which constructed a diagnostic model with a high area under the curve value of 0.9425. We conclude that although urinary sarcosine value has limited potential in the diagnostic algorithm of PCa, urinary metabolomic panel based on GC/MS assay following MAD may potentially become a diagnostic tool for PCa. PMID- 21626194 TI - Analysis of natural red dyes (cochineal) in textiles of historical importance using HPLC and multivariate data analysis. AB - A new analytical approach based on high-performance liquid chromatography with diode array detector (HPLC-DAD) and multivariate data analysis was applied and assessed for analyzing the red dye extracted from cochineal insects, used in precious historical textiles. The most widely used method of analysis involves quantification of specific minor compounds (markers), using HPLC-DAD. However, variation in the cochineal markers concentration, use of aggressive dye extraction methods and poor resolution of HPLC chromatograms can compromise the identification of the precise insect species used in the textiles. In this study, a soft extraction method combined with a new dye recovery treatment was developed, capable of yielding HPLC chromatograms with good resolution, for the first time, for historical cochineal-dyed textiles. After principal components analysis (PCA) and mass spectrometry (MS), it was possible to identify the cochineal species used in these textiles, in contrast to the accepted method of analysis. In order to compare both methodologies, 7 cochineal species and 63 historical cochineal insect specimens were analyzed using the two approaches, and then compared with the results for 15 historical textiles in order to assess their applicability to real complex samples. The methodology developed here was shown to provide more accurate and consistent information than the traditional method. Almost all of the historical textiles were dyed with Porphyrophora sp. insects. These results emphasize the importance of adopting the proposed methodology for future research on cochineal (and related red dyes). Mild extraction methods and HPLC-DAD/MS(n) analysis yield distinctive profiles, which, in combination with a PCA reference database, are a powerful tool for identifying red insect dyes. PMID- 21626195 TI - A new aminopeptidase inhibitor from Streptomyces strain HCCB10043 found by UPLC MS. AB - The diversity of microbial metabolites has been of interest and concern for a long time, yet a suitable method for discovering these is still unavailable. In the work discussed in this report, ultra-performance liquid chromatography coupled with tandem quadrupole and time of flight high-resolution mass spectrometry (UPLC-Q-TOF-HRMS), with MS data analysis, was set up to study the metabolites of Streptomyces strain HCCB10043. It was found that besides antibacterial substances (A21978C complex) and two anti-aminopeptidase compounds (valistatin and bestatin), this strain can produce a new aminopeptidase inhibitor, identified as 3-amino-2-hydroxy-4-phenylbutanoylvalylisoleucine. This new compound had greater activity than valistatin or bestatin in aminopeptidase N (APN) inhibition assay. The results proved that combination of UPLC-Q-TOF-MS analysis and classic purification and identification steps as complementary strategies can provide a method with high reliability for research on microbial secondary metabolites. Furthermore, it has shown that the study of secondary metabolic profiling might be the key to discovering new drugs. PMID- 21626196 TI - Study of OSEM with different subsets in grating-based X-ray differential phase contrast imaging. AB - Impressive developments in X-ray imaging are associated with X-ray phase contrast computed tomography based on grating interferometry, a technique that provides increased contrast compared with conventional absorption-based imaging. A new "single-step" method capable of separating phase information from other contributions has been recently proposed. This approach not only simplifies data acquisition procedures, but, compared with the existing phase step approach, significantly reduces the dose delivered to a sample. However, the image reconstruction procedure is more demanding than for traditional methods and new algorithms have to be developed to take advantage of the "single-step" method. In the work discussed in this paper, a fast iterative image reconstruction method named OSEM (ordered subsets expectation maximization) was applied to experimental data to evaluate its performance and range of applicability. The OSEM algorithm with different subsets was also characterized by comparison of reconstruction image quality and convergence speed. Computer simulations and experimental results confirm the reliability of this new algorithm for phase-contrast computed tomography applications. Compared with the traditional filtered back projection algorithm, in particular in the presence of a noisy acquisition, it furnishes better images at a higher spatial resolution and with lower noise. We emphasize that the method is highly compatible with future X-ray phase contrast imaging clinical applications. PMID- 21626197 TI - [Clinical evaluation of PFNA(r) and relationship between the tip-apex distance and mechanical failure]. AB - The incidence of trochanteric fractures is increasing in Europe, and the economic impact and mortality is high. The aim of the study was to evaluate the PFNA(r) (proximal femoral nail antirotation) with respect to its clinical use and mechanical complications.All patients with a trochanteric fracture who had been treated with a PFNA(r) between 12/2004 and 12/2007 were identified and analysed regarding complications and radiological findings. The study included 195 patients; 61.2% of the patients were classified as Singh I und II. The mean duration of surgery was 57 min. In ten cases (5.1%) the blade migrated, four cases (2.1%) showed blade cut out and in one case the nail broke (0.5%). The mean TAD was 26.7 mm, in cases of cut out 41.3 mm and in blade migrations 38.6 mm. No failure could be documented when the TAD was less then 30 mm. There is a strong relationship between increasing TAD and mechanical failure (P<0.001); 84.6% of the patients have been followed up, and 30.2% died in the follow-up period.The PFNA(r) is an easy-to-use implant for the treatment of stable and instable proximal femur fractures. Mechanical failure depends on the TAD. PMID- 21626198 TI - Genetic architecture of body weight, condition factor and age of sexual maturation in Icelandic Arctic charr (Salvelinus alpinus). AB - The high commercial value from the aquaculture of salmonid fishes has prompted many studies into the genetic architecture of complex traits and the need to identify genomic regions that have repeatable associations with trait variation both within and among species. We searched for quantitative trait loci (QTL) for body weight (BW), condition factor (CF) and age of sexual maturation (MAT) in families of Arctic charr (Salvelinus alpinus) from an Icelandic breeding program. QTL with genome-wide significance were detected for each trait on multiple Arctic charr (AC) linkage groups (BW: AC-4, AC-20; CF: AC-7, AC-20, AC-23, AC-36; MAT: AC-13/34, AC-39). In addition to the genome-wide significant QTL for both BW and CF on AC-20, linkage groups AC-4, AC-7, AC-8, and AC-16 contain QTL for both BW and CF with chromosome-wide significance. These regions had effects (albeit weaker) on MAT with the exception of the region on AC-8. Comparisons with a North American cultured strain of Arctic charr, as well as North American populations of Atlantic salmon (Salmo salar), and rainbow trout (Oncorhynchus mykiss), reveal some conservation in QTL location and structure, particularly with respect to the joint associations of QTL influencing BW and CF. The detection of some differences in genetic architecture between the two aquaculture strains of Arctic charr may be reflective of the differential evolutionary histories experienced by these fishes, and illustrates the importance of including different strains to investigate genetic variation in a species where the intent is to use that variation in selective breeding programs. PMID- 21626199 TI - Current consequences of "Boolean" intuitions. PMID- 21626202 TI - Four-year follow-up of chronic neuronopathic Gaucher disease in Europeans using a modified severity scoring tool. AB - In 2007, the European Task Force for neuronopathic Gaucher disease (NGD) published a review of 55 patients across four countries. Although some observations were possible, analysis was difficult due to the absence of a systematic way of assessing patients. In response to this, a Severity Scoring Tool (SST) was devised to offer a systematic means of assessing the neurological presentation seen. The SST has been modified (mSST) and is a valid tool for monitoring neurological progression. This review describes disease status and progression of neurological manifestations in a cohort of 39 chronic NGD patients across three European countries over a period of 4 years, using the mSST. PMID- 21626203 TI - Reproducibility of fetal ductus venosus blood flow velocity waveforms during first stage of labor. AB - OBJECTIVES: Fetal ductus venosus (DV) blood flow velocity waveforms are significantly altered during contractions in first stage of labor. We have evaluated the reproducibility of these waveforms during and between contractions. METHODS: 90 women between 37th and 41st week of gestation were included in the study. The measurements of the DV were obtained by three different investigators during and between contractions. The pulsatility index for veins (PIV), peak velocity index for veins (PVIV), and fetal heart rate (FHR) were calculated offline. Furthermore, differences in PVIV (PVIVdiff) and PIV (PIVdiff) and a mean of FHR (FHRmean) were calculated. Statistical analysis was used to verify differences in the PIV, PVIV and FHR during and between contractions of each subgroup as well as for the PVIVdiff, PIVdiff and FHRmean between the subgroups. RESULTS: Investigator 1 examined 49 patients (group 1); investigator 2, 17 patients (group 2) and investigator 3, 24 patients (group 3), respectively. In all subgroups, there was a significant difference for the PVIV and PIV during and between contractions, but not for FHR. There was a correlation between gestational age and PVIVdiff, PIVdiff and FHRmean as well as for maternal age and PVIVdiff and PIVdiff. PVIVdiff and PIVdiff showed significant differences in group 1 compared to groups 2 and 3 (p < 0.001), but not between groups 2 and 3. CONCLUSIONS: Significant differences in the measurements in the three subgroups have been revealed, which may be due to different timing of measurement during the contraction. For future studies further standardization of the measurement protocol should be established. PMID- 21626204 TI - High glucose induced rat aorta vascular smooth muscle cell oxidative injury: involvement of protein tyrosine nitration. AB - The alteration and further damage of vascular smooth muscle function have been implicated in the development of vascular complications and diabetes. Little is known about protein tyrosine nitration in vascular smooth muscle cell injury induced by high glucose. In this article, vascular smooth muscle cell was exposed to 30 and 40 mM high glucose for 72 h, and then the cell injury in vascular smooth muscle cell induced by high glucose was studied. It was found that high glucose stimulated vascular smooth muscle cell injury in a dose-dependent manner, including decreasing intracellular and extracellular glutathione contents, increasing malondialdehyde and intracellular reactive oxygen species content, increasing the production of nitric oxide (increased nitrite content in cell and medium), as well as increasing protein tyrosine nitration. By comparing protein tyrosine nitration induced by high glucose conditions and extrinsic factors (hemin-nitrite-glucose oxidase system and 3-morpholinosydnonimine), it may be speculated that protein is nitrated selectively, and specific protein tyrosine nitration is involved in diabetic vascular complications. PMID- 21626205 TI - Age-related changes in xanthine oxidase activity and lipid peroxidation, as well as in the correlation between both parameters, in plasma and several organs from female mice. AB - Xanthine oxidase, a purine catabolism enzyme, has been implicated as an important source of oxidant production and plays an essential role in several inflammatory and oxidative stress-related diseases. It is known that the increasing levels of oxidants cause the chronic oxidative stress characteristic of the ageing process. The aim of the present work was to determine the changes in xanthine oxidase activity and oxidative damage to lipids in several organs (liver, kidney, spleen, lung and two different brain areas, namely cerebral cortex and brainstem) and plasma from two different age groups of BALB/c female mice: adult (7-month-old) and old (18-month-old) mice, as well as to analyse the possible correlation between both parameters. Xanthine oxidase activity was significantly increased in liver, cerebral cortex and plasma from old mice in comparison with adults. Similar results were obtained in the lipid peroxidation levels, in which old mice showed a high increment in liver and cerebral cortex. Moreover, the results show a significant and positive correlation between xanthine oxidase activity and lipid peroxidation levels in cerebral cortex. The age-related increase in the xanthine oxidase activity and lipid peroxidation in liver and cerebral cortex of mice seems to suggest that the xanthine oxidase plays a role in the acceleration of the oxidative damage in these organs with age and its possible contribution to the pathophysiological changes associated to the process of ageing. PMID- 21626207 TI - Production of xylanolytic enzymes by Aspergillus terricola in stirred tank and airlift tower loop bioreactors. AB - Fungi producing high xylanase levels have attracted considerable attention because of their potential industrial applications. Batch cultivations of Aspergillus terricola fungus were evaluated in stirred tank and airlift bioreactors, by using wheat bran particles suspended in the cultivation medium as substrate for xylanase and beta-xylosidase production. In the stirred tank bioreactor, in physical conditions of 30 degrees C, 300 rpm, and aeration of 1 vvm (1 l min-1), with direct inoculation of fungal spores, 7,475 U l-1 xylanase was obtained after 36 h of operation, remaining constant after 24 h. In the absence of air injection in the stirred tank reactor, limited xylanase production was observed (final concentration 740 U l-1). When the fermentation process was realized in the airlift bioreactor, xylanase production was higher than that observed in the stirred tank bioreactor, being 9,265 U l-1 at 0.07 vvm (0.4 l min 1) and 12,845 U l-1 at 0.17 vvm (1 l min-1) aeration rate. PMID- 21626208 TI - Engineering Saccharomyces cerevisiae to produce feruloyl esterase for the release of ferulic acid from switchgrass. AB - The Aspergillus niger feruloyl esterase gene (faeA) was cloned into Saccharomyces cerevisiae via a yeast expression vector, resulting in efficient expression and secretion of the enzyme in the medium with a yield of ~2 mg/l. The recombinant enzyme was purified to homogeneity by anion-exchange and hydrophobic interaction chromatography. The specific activity was determined to be 8,200 U/MUg (pH 6.5, 20 degrees C, 3.5 mM 4-nitrophenyl ferulate). The protein had a correct N terminal sequence of ASTQGISEDLY, indicating that the signal peptide was properly processed. The FAE exhibited an optimum pH of 6-7 and operated optimally at 50 degrees C using ground switchgrass as the substrate. The yeast clone was demonstrated to catalyze the release of ferulic acid continuously from switchgrass in YNB medium at 30 degrees C. This work represents the first report on engineering yeast for the breakdown of ferulic acid crosslink to facilitate consolidated bioprocessing. PMID- 21626209 TI - Intrinsic and inducible resistance to hydrogen peroxide in Bifidobacterium species. AB - Interest in, and use of, bifidobacteria as a probiotic delivered in functional foods has increased dramatically in recent years. As a result of their anaerobic nature, oxidative stress can pose a major challenge to maintaining viability of bifidobacteria during functional food storage. To better understand the oxidative stress response in two industrially important bifidobacteria species, we examined the response of three strains of B. longum and three strains of B. animalis subsp. lactis to hydrogen peroxide (H2O2). Each strain was exposed to a range of H2O2 concentrations (0-10 mM) to evaluate and compare intrinsic resistance to H2O2. Next, strains were tested for the presence of an inducible oxidative stress response by exposure to a sublethal H2O2 concentration for 20 or 60 min followed by challenge at a lethal H2O2 concentration. Results showed B. longum subsp. infantis ATCC 15697 had the highest level of intrinsic H2O2 resistance of all strains tested and B. animalis subsp. lactis BL-04 had the highest resistance among B. lactis strains. Inducible H2O2 resistance was detected in four strains, B. longum NCC2705, B. longum D2957, B. lactis RH-1, and B. lactis BL-04. Other strains showed either no difference or increased sensitivity to H2O2 after induction treatments. These data indicate that intrinsic and inducible resistance to hydrogen peroxide is strain specific in B. longum and B. lactis and suggest that for some strains, sublethal H2O2 treatments might help increase cell resistance to oxidative damage during production and storage of probiotic containing foods. PMID- 21626210 TI - Chloroplasts move towards the nearest anticlinal walls under dark condition. AB - Chloroplasts change their intracellular positions in response to their light environment. Under darkness, chloroplasts assume special positions that are different from those under light conditions. Here, we analyzed chloroplast dark positioning using Adiantum capillus-veneris gametophyte cells. When chloroplasts were transferred into darkness, during the first 1-5 h, they moved towards the anticlinal cell walls bordering the adjacent cells rather rapidly. Then, they slowed down and accumulated at the anticlinal walls gradually over the following 24-36 h. The chloroplast movements could be roughly classified into two different categories: initial rapid straight movement and later, slow staggering movement. When the chloroplast accumulation response was induced in dark-adapted cells by partial cell irradiation with a microbeam targeted to the center of the cells, chloroplasts moved towards the beam spot from the anticlinal walls. However, when the microbeam was switched off, they moved to the nearest anticlinal walls and not to their original positions if they were not the closest, indicating that they know the direction of the nearest anticlinal wall and do not have particular areas that they migrate to during dark positioning. PMID- 21626211 TI - Origin and evolution of genes related to ABA metabolism and its signaling pathways. AB - Since plants cannot move to avoid stress, they have sophisticated acclimation mechanisms against a variety of abiotic stresses. The phytohormone abscisic acid (ABA) plays essential roles in abiotic stress tolerances in land plants. Therefore, it is interesting to address the evolutionary origins of ABA metabolism and its signaling pathways in land plants. Here, we focused on 48 ABA related Arabidopsis thaliana genes with 11 protein functions, and generated 11 orthologous clusters of ABA-related genes from A. thaliana, Arabidopsis lyrata, Populus trichocarpa, Oryza sativa, Selaginella moellendorffii, and Physcomitrella patens. Phylogenetic analyses suggested that the common ancestor of these six species possessed most of the key protein functions of ABA-related genes. In two species (A. thaliana and O. sativa), duplicate genes related to ABA signaling pathways contribute to the expression variation in different organs or stress responses. In particular, there is significant expansion of gene families related to ABA in evolutionary periods associated with morphological divergence. Taken together, these results suggest that expansion of the gene families related to ABA signaling pathways may have contributed to the sophisticated stress tolerance mechanisms of higher land plants. PMID- 21626212 TI - AUTOBA: automation of backbone assignment from HN(C)N suite of experiments. AB - Development of efficient strategies and automation represent important milestones of progress in rapid structure determination efforts in proteomics research. In this context, we present here an efficient algorithm named as AUTOBA (Automatic Backbone Assignment) designed to automate the assignment protocol based on HN(C)N suite of experiments. Depending upon the spectral dispersion, the user can record 2D or 3D versions of the experiments for assignment. The algorithm uses as inputs: (i) protein primary sequence and (ii) peak-lists from user defined HN(C)N suite of experiments. In the end, one gets H(N), (15)N, C(alpha) and C' assignments (in common BMRB format) for the individual residues along the polypeptide chain. The success of the algorithm has been demonstrated, not only with experimental spectra recorded on two small globular proteins: ubiquitin (76 aa) and M-crystallin (85 aa), but also with simulated spectra of 27 other proteins using assignment data from the BMRB. PMID- 21626213 TI - Protein-protein HADDocking using exclusively pseudocontact shifts. AB - In order to enhance the structure determination process of macromolecular assemblies by NMR, we have implemented long-range pseudocontact shift (PCS) restraints into the data-driven protein docking package HADDOCK. We demonstrate the efficiency of the method on a synthetic, yet realistic case based on the lanthanide-labeled N-terminal epsilon domain of the E. coli DNA polymerase III (epsilon186) in complex with the HOT domain. Docking from the bound form of the two partners is swiftly executed (interface RMSDs < 1 A) even with addition of very large amount of noise, while the conformational changes of the free form still present some challenges (interface RMSDs in a 3.1-3.9 A range for the ten lowest energy complexes). Finally, using exclusively PCS as experimental information, we determine the structure of epsilon186 in complex with the HOT homologue theta subunit of the E. coli DNA polymerase III. PMID- 21626214 TI - A systematic mutagenesis-driven strategy for site-resolved NMR studies of supramolecular assemblies. AB - Obtaining sequence-specific assignments remains a major bottleneck in solution NMR investigations of supramolecular structure, dynamics and interactions. Here we demonstrate that resonance assignment of methyl probes in high molecular weight protein assemblies can be efficiently achieved by combining fast NMR experiments, residue-type-specific isotope-labeling and automated site-directed mutagenesis. The utility of this general and straightforward strategy is demonstrated through the characterization of intermolecular interactions involving a 468-kDa multimeric aminopeptidase, PhTET2. PMID- 21626215 TI - Knowledge-based nonuniform sampling in multidimensional NMR. AB - The full resolution afforded by high-field magnets is rarely realized in the indirect dimensions of multidimensional NMR experiments because of the time cost of uniformly sampling to long evolution times. Emerging methods utilizing nonuniform sampling (NUS) enable high resolution along indirect dimensions by sampling long evolution times without sampling at every multiple of the Nyquist sampling interval. While the earliest NUS approaches matched the decay of sampling density to the decay of the signal envelope, recent approaches based on coupled evolution times attempt to optimize sampling by choosing projection angles that increase the likelihood of resolving closely-spaced resonances. These approaches employ knowledge about chemical shifts to predict optimal projection angles, whereas prior applications of tailored sampling employed only knowledge of the decay rate. In this work we adapt the matched filter approach as a general strategy for knowledge-based nonuniform sampling that can exploit prior knowledge about chemical shifts and is not restricted to sampling projections. Based on several measures of performance, we find that exponentially weighted random sampling (envelope matched sampling) performs better than shift-based sampling (beat matched sampling). While shift-based sampling can yield small advantages in sensitivity, the gains are generally outweighed by diminished robustness. Our observation that more robust sampling schemes are only slightly less sensitive than schemes highly optimized using prior knowledge about chemical shifts has broad implications for any multidimensional NMR study employing NUS. The results derived from simulated data are demonstrated with a sample application to PfPMT, the phosphoethanolamine methyltransferase of the human malaria parasite Plasmodium falciparum. PMID- 21626216 TI - Elucidating slow binding kinetics of a protein without observable bound resonances by longitudinal relaxation NMR spectroscopy. AB - We developed a new method to elucidate the binding kinetics k(on) and k(off), and the dissociation constant K(D) (=k(off)/k(on)), of protein-protein interactions without observable bound resonances of the protein of interest due to high molecular weight in a complex with a large target protein. In our method, k(on) and k(off) rates are calculated from the analysis of longitudinal relaxation rates of free resonances measured for multiple samples containing different concentration ratios of (15)N-labeled protein and substoichiometric amounts of the target protein. The method is applicable to interactions that cannot be analyzed by relaxation dispersion spectroscopy due to slow interactions on millisecond to second timescale and/or minimal conformational (chemical shift) change upon binding. We applied the method to binding of the B1 domain of protein G (GB1) to immunoglobulin G, and derived the binding kinetics despite the absence of observable bound GB1 resonances. PMID- 21626217 TI - Polymorphisms of the beta adrenergic receptor predict left ventricular remodeling following acute myocardial infarction. AB - PURPOSE: Prior studies demonstrate an association between specific beta adrenergic receptor (beta-AR) polymorphisms and clinical outcomes in patients with chronic heart failure and following acute coronary syndromes. The underlying mechanism may be due to differences in left ventricular remodeling. This study was undertaken to explore the relationship between LV remodeling after myocardial infarction and polymorphisms in the cardiac beta1-AR and beta2-AR genes. METHODS: After first ST-segment elevation myocardial infarction (STEMI), 122 patients on chronic beta1 receptor antagonist therapy underwent baseline and 6-month LV volume evaluation. We assessed the relationships between changes in LV volumes and the polymorphisms in beta1-AR, beta1-Arg389Gly and beta1-Ser49Gly, and in beta2-AR, beta2-Gly16Arg and beta2-Gln27Glu. RESULTS: We found that patients homozygous for the beta2-Glu27 variant were 5.2 times more likely to be in the group with the highest end systolic volume (ESV) progression (OR 5.2, 95%CI 1.4 19.0). They were also more likely to have the largest progression of end diastolic volume (EDV) and decrease in LV ejection fraction (LVEF). For those with baseline LV dysfunction, being homozygous for Arg at amino acid position 389 in beta1-AR was associated with decreases in ESV (-46 mL, CI -3.1, -88) and EDV ( 40 mL, CI -1.1, -79) and an increase in LVEF (11%, CI 0.3, 22). CONCLUSION: We found that polymorphisms of the beta1-AR and beta2-AR genes are associated with differential LV remodeling in patients treated with a beta1 receptor antagonist following STEMI. PMID- 21626218 TI - Effects of omega-3 fatty acid for sudden cardiac death prevention in patients with cardiovascular disease: a contemporary meta-analysis of randomized, controlled trials. AB - PURPOSE: Experimental and epidemiological studies suggest that omega-3 fatty acids have an antiarrhythmic effect. However, evidence from randomized controlled trials (RCTs) for prevention of sudden cardiac death (SCD) remains controversial. This study sought to evaluate the efficacy of omega-3 fatty acids for secondary prevention of SCD in patients with cardiovascular disease (CVD) in the era of guidelines-based therapy. METHODS: We conducted a PubMed/EMBASE/CENTRAL search for RCTs evaluating omega-3 fatty acids for CVD secondary prevention with at least 6 months follow-up and with data on SCD. Primary outcome was SCD. Secondary outcomes were cardiovascular mortality and all-cause mortality. RESULTS: Ten randomized controlled trials were identified evaluating a total of 33,429 patients with CVD. In patients with guidelines-adjusted therapy, omega-3 fatty acids did not reduce the risk ratio (RR) of SCD (RR:0.96; 95% CI: 0.84-1.10). In patients with non- guidelines-adjusted therapy, omega-3 fatty acids reduced the RR of SCD (RR: 0.64; 95% CI: 0.51-0.80). Overall, RR for cardiac death and all cause mortality were 0.81 (95% CI: 0.69-0.95) and 0.89 (95% CI: 0.79-1.01), respectively. CONCLUSIONS: In the era of guidelines-adjusted treatment for CVD secondary prevention, omega-3 fatty acids do not appear to reduce SCD. PMID- 21626219 TI - More education, less administration: reflections of neuroimagers' attitudes to ethics through the qualitative looking glass. AB - In follow-up to a large-scale ethics survey of neuroscientists whose research involves neuroimaging, brain stimulation and imaging genetics, we conducted focus groups and interviews to explore their sense of responsibility about integrating ethics into neuroimaging and readiness to adopt new ethics strategies as part of their research. Safety, trust and virtue were key motivators for incorporating ethics into neuroimaging research. Managing incidental findings emerged as a predominant daily challenge for faculty, while student reports focused on the malleability of neuroimaging data and scientific integrity. The most frequently cited barrier was time and administrative burden associated with the ethics review process. Lack of scholarly training in ethics also emerged as a major barrier. Participants constructively offered remedies to these challenges: development and dissemination of best practices and standardized ethics review for minimally invasive neuroimaging protocols. Students in particular, urged changes to curricula to include early, focused training in ethics. PMID- 21626220 TI - Acetyl salicylic acid treatment in neonatal Bartter syndrome--a commentary letter. PMID- 21626221 TI - Intensive hemodialysis for cardiomyopathy associated with end-stage renal disease. AB - Heart and kidney dysfunction often coexist, and increasing evidence supports the interaction of these two organs, as demonstrated by the clinical condition known as cardiorenal syndrome (CRS). We report a pediatric patient with end-stage renal disease (ESRD) who developed a dilated cardiomyopathy and decompensated heart failure after undergoing unilateral nephrectomy and while on maintenance peritoneal dialysis. He showed marked improvement in his cardiac function with the addition of intensive hemodialysis. We discuss the pathophysiology of cardiorenal syndrome in patients with ESRD and suggest that intensive dialysis may be an effective therapy for this condition. PMID- 21626222 TI - Cyclosporine A vs. methylprednisolone for Henoch-Schonlein nephritis: a randomized trial. AB - Knowledge about how to treat severe Henoch-Schonlein nephritis (HSN) is scarce. The aim of our study is to compare cyclosporine A (CyA) and methylprednisolone pulses (MP) in the treatment of severe HSN. Out of 24 pediatric HSN patients with nephrotic-range proteinuria or crescentic HSN in kidney biopsy, seven were randomized to receive CyA for 12 months at an initial dose of 5 mg/kg and eight to receive 3 MP pulses of 30 mg/kg followed by prednisone for 4 months. The other nine patients received identical treatment without randomization. Kidney biopsies were performed at inclusion and after 2 years. The primary outcomes were the duration of proteinuria and hematuria, estimated glomerular filtration rate, and renal biopsy histology. All the 11 CyA-treated patients achieved resolution of nephrotic-range proteinuria within 3 months, while the MP-group response was slower, and in 6/13 was not achieved with the initial treatment. Additional immunosuppressive treatment was needed in none of the CyA-treated patients but in six patients treated with MP (difference in proportion 46%, p = 0.008). The 2 year control biopsies were similarly improved in both groups. After mean 6.1 years (2.2-10.4 years), 16 patients (eight CyA, eight MP) had no renal symptoms and six (three CyA, three MP) had persistent nephropathy but normal renal function. One MP-treated patient had reduced renal function and another had developed ESRD and received a renal transplant. CyA gave a 100% resolution of nephrotic-range proteinuria and a 100% renal survival rate without additional therapy after a mean follow-up of 6 years. Treatment of HSN with CyA is efficacious, safe and not inferior to MP. PMID- 21626223 TI - The impact of pediatric nephrotic syndrome on families. AB - The objective of our study was to assess the psychologic and economic effects of pediatric nephrotic syndrome (NS) on caregivers. Caregivers of 50 children with NS were compared with a control group of 50 families of children with minor illnesses attending the same outpatient facility. Beck's Depression Inventory (BDI) IA was used to assess the mental status of the primary caregiver. The socioeconomic status of the family was assessed using the modified Kuppuswamy scale. Expenditure for the illness was calculated during parent interviews. The difference between groups was analyzed using analysis of variance (ANOVA) and Duncan's multiple range test. BDI scores signified moderate to severe depression in 48% of NS caregivers compared with 12% controls. The mean BDI score was significantly higher in NS caregivers, correlating positively with disease severity and negatively with socioeconomic status. Expenditure for disease also was significantly higher in families with NS patients, varying between 30% and 60% of monthly income depending on disease severity compared with 6.9% in controls. In 10% of NS families, it was more than total income, forcing families to break into savings or go into debt. Although pediatric NS most commonly has an excellent long-term outcome, it causes significant mental and economic stress on families. Severe forms should be categorized as a chronic illness and be eligible for disability benefits and subsidized travel and medical care. Establishing support groups and supportive care at local levels would help reduce the burden on families of patients wtih NS. PMID- 21626224 TI - A framework to establish a mentoring programme in surgery. AB - INTRODUCTION: Mentoring programmes help to facilitate the process of continuous professional development in surgery, providing an organizational structure around a mentor-mentee relationship which helps to develop the mentee. The lack of guidelines outlining how to set up such mentoring programmes, the fragmented inter-relationships of existing schemes and the lack of a unified strategy for their implementation are obstacles to the creation of such initiatives within many surgical departments. METHODS: We draw upon previous research, the experiences of certain authors and our own reflections to identify the key features of a surgical mentoring programme. RESULTS: We propose a ten step process which aims to encourage the development of formalised mentoring programmes in surgery. CONCLUSION: This outline may improve the delivery and effectiveness of mentoring programmes, which may ultimately enhance surgical training and hence quality of patient care. PMID- 21626225 TI - Is the severity of adolescent conduct disorder associated with the level of nicotine dependence? AB - Our aim was to investigate the relationship between the number of Conduct Disorder (CD) symptoms and specific DSM-IV symptom subscales (i.e., aggression towards people and animals, destruction of property, deceitfulness or theft, and serious violations of rules) and nicotine dependence (ND) among adolescent psychiatric inpatients in Finland. A total of 171 adolescents with CD (99 boys and 72 girls; age 12-17 years) were admitted to inpatient psychiatric hospitalization between April 2001 and March 2006. Information on their psychiatric DSM-IV diagnoses was obtained using the Schedule for Affective Disorder and Schizophrenia for School-Age Children (K-SADS-PL). The level of ND was assessed with the modified Fagerstrom Tolerance Questionnaire (mFTQ). The total number of CD symptoms correlated with the level of ND among both the boys (adj. r = 0.31, p = 0.002) and the girls (adj. r = 0.324, p = 0.006). For the boys, the number of conduct symptoms correlated with the level of ND on all subscales except for aggression, while the only statistically significant correlation for the girls was found with the deceitfulness or theft subscale. A comorbid substance use disorder was statistically significant and associated with a high level of ND among the boys (p < 0.001) and the girls (p = 0.019). Our results suggest that both in adolescent boys and girls, the greater the number of CD symptoms, the higher the level of ND. Future studies are needed on the relationship between environmental factors, non-aggressive CD symptoms and the development of ND among adolescents with CD. PMID- 21626226 TI - Diurnal variations in arousal: a naturalistic heart rate study in children with ADHD. AB - Previous studies suggest an altered circadian regulation of arousal in children with attention-deficit hyperactivity disorder (ADHD) as measured by activity, circadian preference, and sleep-wake patterns. Although heart rate is an important measure to evaluate arousal profiles, to date it is unknown whether 24 h heart rate patterns differentiate between children with and without ADHD. In this study, 24-h heart rate data were collected in 30 non-medicated children with ADHD (aged 6-11) and 30 sex-, class-, and age-matched normal controls in their naturalistic home and school setting, during 5 days. Simultaneously, 24-h activity patterns were registered. Confounding effects of demographic variables (e.g., age, sex, BMI, pubertal stage) and comorbid internalizing and externalizing problems on heart rate levels were additionally assessed. Longitudinal analysis showed that heart rate levels were overall higher in the ADHD group (p < 0.01)--with the largest effects during afternoon and night--in a model controlling for age. Other factors did not significantly contribute to variations in heart rate levels. Compared to controls, children with ADHD showed higher activity levels during daytime (especially early afternoon), but not during nighttime (p < 0.05). Post hoc analyses showed that environmental effects might influence daytime variations. Findings suggest an autonomic imbalance in children with ADHD as compared to controls, with higher heart rate levels in the ADHD group. Nighttime tachycardia in this group could not be explained by nighttime activity levels or comorbid externalizing/internalizing problems. Further research on autonomic functioning in ADHD is recommended because of the major impact of higher resting heart rate on health outcomes. PMID- 21626228 TI - High prevalence of cholelithiasis in primary hyperparathyroidism: a retrospective analysis of 120 cases. PMID- 21626227 TI - Auditory nerve excitation via a non-traveling wave mode of basilar membrane motion. AB - Basilar membrane (BM) motion and auditory nerve fiber (ANF) tuning are generally very similar, but the ANF had appeared to be unresponsive to a plateau mode of BM motion that occurs at frequencies above an ANF's characteristic frequency (CF). We recorded ANF responses from the gerbil, concentrating on this supra-CF region. We observed a supra-CF plateau in ANF responses at high stimulus level, indicating that the plateau mode of BM motion can be excitatory. PMID- 21626229 TI - Factors associated with operative recurrence early after resection for Crohn's disease. AB - INTRODUCTION: Some Crohn's disease (CD) patients develop rapid disease recurrence requiring reoperation. Identification of factors associated with early operative recurrence of CD may help risk-stratify patients and prevent recurrence. METHODS: Prospectively collected data of CD patients undergoing bowel resection for CD with unequivocal evidence of recurrence at reoperation were retrieved. Patients with earlier recurrence (less than median time of recurrence of study cohort) were compared with those who developed later recurrence (greater than median time of recurrence) for patient and disease characteristics and risk factors for recurrence. A multivariate logistic regression model was performed to identify factors associated with earlier operative recurrence. RESULTS: Sixty-nine patients (45 female, 24 male) met the inclusion criteria. Median time to reoperation was 38 months (range, 3.3-236 months). One hundred six reoperations in the 69 patients were for abscess/fistula/perforation (n = 45), stricture/stenosis (n = 41), inflammation (n = 17), bleeding (n = 2), and dysplasia (n = 1). Factors associated with early rather than late reoperation included behavior of disease (stricturing, odds ratio (OR) 12.1; confidence interval (CI), 1.8-80.9; penetrating OR, 9.9; CI, 1.4-67.9 rather than nonstricturing nonpenetrating) and the development of postoperative complications at previous surgery (OR, 12.1; CI, 1.2-126.6). CONCLUSION: Earlier recurrence of CD requiring reoperation is associated with specific disease and potentially modifiable operation-related factors such as postoperative complications, i.e., anastomotic leak or intraabdominal abscess. Strategies to reduce recurrence in such patients include the identification of factors that may reduce postoperative complications. PMID- 21626230 TI - Absence of mutation at the 5'-upstream promoter region of the TPM4 gene from cardiac mutant axolotl (Ambystoma mexicanum). AB - Tropomyosins are a family of actin-binding proteins that show cell-specific diversity by a combination of multiple genes and alternative RNA splicing. Of the 4 different tropomyosin genes, TPM4 plays a pivotal role in myofibrillogenesis as well as cardiac contractility in amphibians. In this study, we amplified and sequenced the upstream regulatory region of the TPM4 gene from both normal and mutant axolotl hearts. To identify the cis-elements that are essential for the expression of the TPM4, we created various deletion mutants of the TPM4 promoter DNA, inserted the deleted segments into PGL3 vector, and performed promoter reporter assay using luciferase as the reporter gene. Comparison of sequences of the promoter region of the TPM4 gene from normal and mutant axolotl revealed no mutations in the promoter sequence of the mutant TPM4 gene. CArG box elements that are generally involved in controlling the expression of several other muscle specific gene promoters were not found in the upstream regulatory region of the TPM4 gene. In deletion experiments, loss of activity of the reporter gene was noted upon deletion which was then restored upon further deletion suggesting the presence of both positive and negative cis-elements in the upstream regulatory region of the TPM4 gene. We believe that this is the first axolotl promoter that has ever been cloned and studied with clear evidence that it functions in mammalian cell lines. Although striated muscle-specific cis-acting elements are absent from the promoter region of TPM4 gene, our results suggest the presence of positive and negative cis-elements in the promoter region, which in conjunction with positive and negative trans-elements may be involved in regulating the expression of TPM4 gene in a tissue-specific manner. PMID- 21626231 TI - Memory and metamemory for inverted words: illusions of competency and desirable difficulties. AB - Research regarding how people monitor their learning has shown that ease of processing strongly guides people's judgments of learning (JOLs). However, the desirable difficulties concept (Bjork, 1994) suggests that studying information that is less fluent can result in greater learning. Currently, it is unclear whether people are aware of the potential benefits of desirable difficulties during learning. To address this, in Experiment 1, participants studied inverted and upright words and also made JOLs. While participants' JOLs did not differ for inverted and upright words, recall was greater for inverted words. Experiment 2 used several study-test cycles in which participants could potentially learn about the beneficial effects of processing inverted words with task experience, and similar results were obtained. Thus, reading inverted words requires processing that enhances recall, but memory predictions do not differentiate between upright and inverted words. We interpret these results in terms of processing fluency, desirable difficulties, and theories of metacognitive monitoring. PMID- 21626232 TI - Discriminatory mass de-housing and low-weight births: scales of geography, time, and level. AB - Struening et al.1 demonstrated a widening disparity of low birthweight (LOB) rates among New York City health areas from 1980-1986, clearly a dynamic process. In contrast, the New York City Department of Health reported static citywide LOB rate in 1988-2008.2 Struening et al.1 is extended here at the health district level with mapping and regression analyses. Additionally, birthweight data are reported for babies born in 1998-2001 to a group of African-American and Dominican women in Upper Manhattan. The data reported in this paper indicate that both fetal programming of the mother herself (life course model) and stress during or shortly before pregnancy may play a role in LOB. Current stress may arise from past events. Intergenerational effects, thus, could arise from stresses on the grandmother and their residual impacts on the mother as well as new stresses on the mother as an adult. The average weight of babies born to the Upper Manhattan mothers who were born in 1970-1974 was 3,466 g, with 1.6% below 2,500 g; that of babies of mothers born in 1975-1979, 3,320 g, with 6% below 2,500 g. The latter group was born during the 1975-1979 housing destruction. Intergenerational impacts of that event may be reflected in this elevated rate of LOB. Health district maps of LOB incidence ranges show improvement from 1990-2000 and then deterioration in 2005 and 2008. Bivariate regressions of socioeconomic (SE) factors and LOB incidence showed many strong associations in 1990; but by 2000, the number and strength of these associations declined. In 1990, 2000, and 2008, black segregation was the SE factor most strongly associated with LOB. Black segregation and murder rate explained about 85% of the pattern of 1990 LOB. Regressing the 1970-1980 percent population change against the SE factors showed effects even in 2000. The 1990 murder rate and 1989 percentage of public assistance explained over half the 2008 LOB incidence pattern. The housing destruction of the 1970s continued to influence LOB incidence indirectly in 2008. The ability of community and individual to cope with current stressors may hinge on resilience status, which is shaped by past events and circumstances. The present interacts with the past in many ways. Serial displacement exemplifies this interaction of immense importance to public health. PMID- 21626233 TI - Old issues and new perspectives on prostate cancer hormonal therapy: the molecular substratum. AB - Secondary hormonal therapy is a treatment option in patients with castration resistant prostate cancer (CRPC); however, it is underutilized and is room for optimization and improvement. In this context, androgen receptor (AR) is the Achilles' heel, being critically important and various mechanisms ranging from receptor mutations to secondary signaling pathways are responsible for some of the biological heterogeneity, demanding a multimodal approach. A comprehensive review of the peer-reviewed literature is performed on the topic of molecular mechanisms supporting secondary hormonal therapies, including expanded alternative hormonal therapies for CRPC. Essential concepts in clinical treatment of patients with progression on primary hormonal therapy are maintaining the castrate state, accounting for the intermittency phenomenon and sequentially using oral antiandrogens and adrenolytics heading to androgen depletion microenvironment. Survival prolongation, pain relief or measurable improvement in tumor-related symptoms should be persecuted and are considered to be a tangible benefit of obvious worth to the patient. Understanding the underlying molecular substratum is of paramount importance to hormonal therapy optimization in this context once current androgen-depletion strategies are incomplete, and residual androgens as well as alternative routes contribute to sustained AR activity and disease progression to a lethal phenotype. One or many mechanisms may be playing a role, even within the same patient and lastly are potential targets for treatment. Five fundamental mechanisms mediated through the AR to promote tumor growth (three of which depend on ligand signaling) added to the stem cell pathway must be recognized in CRPC. They are persistence of intratumoral androgens as a result of in situ steroidogenesis or adrenal source; AR mutations that allow promiscuous activation by otherwise nonsignaling ligands; wild-type AR gene amplification; alterations in AR coactivator-to-corepressor ratio that impact transcription; outlaw AR pathways that bypass the need for androgens by signaling through crosstalk with other ligand-bound receptors, cytokines, or transactivation of activated tyrosine kinase receptors in the cytosol. PMID- 21626234 TI - Differential histomechanical response of carotid artery in relation to species and region: mathematical description accounting for elastin and collagen anisotropy. AB - The selection of a mathematical descriptor for the passive arterial mechanical behavior has been long debated in the literature and customarily constrained by lack of pertinent data on the underlying microstructure. Our objective was to analyze the response of carotid artery subjected to inflation/extension with phenomenological and microstructure-based candidate strain-energy functions (SEFs), according to species (rabbit vs. pig) and region (proximal vs. distal). Histological variations among segments were examined, aiming to explicitly relate them with the differential material response. The Fung-type model could not capture the biphasic response alone. Combining a neo-Hookean with a two-fiber family term alleviated this restraint, but force data were poorly captured, while consideration of low-stress anisotropy via a quadratic term allowed improved simulation of both pressure and force data. The best fitting was achieved with the quadratic and Fung-type or four-fiber family SEF. The latter simulated more closely than the two-fiber family the high-stress response, being structurally justified for all artery types, whereas the quadratic term was justified for transitional and muscular arteries exhibiting notable elastin anisotropy. Diagonally arranged fibers were associated with pericellular medial collagen, and circumferentially and longitudinally arranged fibers with medial and adventitial collagen bundles, evidenced by the significant correlations of SEF parameters with quantitative histology. PMID- 21626235 TI - The importance of 'social responsibility' in the promotion of health. AB - The publication of the Report of the International Bioethics Committee of Unesco on Social responsibility and health provides an opportunity to reshape the conceptual framework of the right to health care and its practical implications. The traditional distinctions between negative and positive, civil-political and economic-social, legal and moral rights are to be questioned and probably overcome if the goal is to pursue 'the highest attainable standard of health' as a fundamental human right, that should as such be guaranteed to every human being. What we are called upon to, is the commitment not to exclude now and forever anyone from having access to the 'excellence' of scientific and medical progress. Therefore, the addressees of this 'responsibility' cannot be just the governments and the states within the limits of their 'jurisdiction'. The challenge is to tackle at the same time the social and global determinants of health. PMID- 21626236 TI - BraSto, a Stowaway MITE from Brassica: recently active copies preferentially accumulate in the gene space. AB - We characterized a Brassica miniature inverted repeat transposable element (MITE) from the Stowaway superfamily, designated BraSto (Bra ssica Sto waway). BraSto copy number was assessed using real-time quantitative PCR in the two diploid species B. rapa (genome A) and B. oleracea (genome C) and the corresponding allotetraploid species B. napus (genome AC). Phylogenetic relationships among a set of 131 BraSto copies were then analyzed. BraSto appears to have been only moderately amplified in the Brassica genome and was still active recently with marks of proliferation in both diploid Brassica species, which diverged 3.75 million years ago, but also in the allotetraploid species after reuniting of the two differentiated genomes. We characterized insertion sites for low-divergence BraSto copies among the gene space of the B. rapa genome using bioinformatics approaches. For BraSto copies localized nearby or within genes, we observed frequent associations of BraSto with putative promoters and regulatory regions of genes, but exclusion from coding regions. In addition, BraSto was significantly similar to several Brassica expressed sequence tags (ESTs), including stress induced ESTs. We also demonstrated the enrichment of BraSto sequences in binding sites for transcription factors and other regulatory elements. Our results lead to the question of a role for BraSto in the regulation of gene expression: this putative role, if further confirmed experimentally, would help to obtain a new insight into the significance of MITEs in the functional plant genome. PMID- 21626237 TI - Cytotoxicity of methylsulfonylmethane on gastrointestinal (AGS, HepG2, and KEYSE 30) cancer cell lines. AB - PURPOSE: This study was conducted to assay cytotoxic effects of methylsulfonylmethane (MSM) on gastrointestinal cancer cell lines. METHODS: Human gastric carcinoma (AGS), human hepatocellular carcinoma (HepG2), and human esophageal squamous cell carcinoma (KYSE-30) cancer cell lines were treated by MSM and incubated for 24, 48, and 72 h. Cytotoxicity was examined through MTT, neutral red uptake, and protein measurement assays. Ethidium bromide/acridine orange (EB/AO) staining was used for apoptotic cell detection. A diamidino-2 phenylindole staining method was used to analysis cell cycle by flow cytometry. RESULTS: IC(50) of MSM on AGS, HepG2, and KYSE-30 cell lines were 28.04, 21.87 and 27.98 mg/ml after 72 h, respectively. The EB/AO staining showed an increase in apoptotic cells. Cell cycle analysis showed a significant increase in cell density at G2/M phase. CONCLUSION: MSM had cytotoxic effect on cancer cell lines but HepG2 cell line was more susceptible. This study suggests that MSM may induce cytotoxic effect on gastrointestinal cancer cell lines by apoptosis and cell cycle arrest. PMID- 21626238 TI - Hepato-biliary Cystadenoma with Intraductal Extension: Unusual Cause of Obstructive Jaundice. PMID- 21626239 TI - Person perception informs understanding of cognition during visual search. AB - Does person perception--the impressions we form from watching others--hold clues to the mental states of people engaged in cognitive tasks? We investigated this with a two-phase method: In Phase 1, participants searched on a computer screen (Experiment 1) or in an office (Experiment 2); in Phase 2, other participants rated the searchers' video-recorded behavior. The results showed that blind raters are sensitive to individual differences in search proficiency and search strategy, as well as to environmental factors affecting search difficulty. Also, different behaviors were linked to search success in each setting: Eye movement frequency predicted successful search on a computer screen; head movement frequency predicted search success in an office. In both settings, an active search strategy and positive emotional expressions were linked to search success. These data indicate that person perception informs cognition beyond the scope of performance measures, offering the potential for new measurements of cognition that are both rich and unobtrusive. PMID- 21626242 TI - Still in search of bisexual sexual arousal: comment on Cerny and Janssen (2011). PMID- 21626240 TI - Family members' experiences of "wait and see" as a communication strategy in end of-life decisions. AB - PURPOSE: The aim of this study is to examine family members' experiences of end of-life decision-making processes in Norwegian intensive care units (ICUs) to ascertain the degree to which they felt included in the decision-making process and whether they received necessary information. Were they asked about the patient's preferences, and how did they view their role as family members in the decision-making process? METHODS: A constructivist interpretive approach to the grounded theory method of qualitative research was employed with interviews of 27 bereaved family members of former ICU patients 3-12 months after the patient's death. RESULTS: The core finding is that relatives want a more active role in end of-life decision-making in order to communicate the patient's wishes. However, many consider their role to be unclear, and few study participants experienced shared decision-making. The clinician's expression "wait and see" hides and delays the communication of honest and clear information. When physicians finally address their decision, there is no time for family participation. Our results also indicate that nurses should be more involved in family-physician communication. CONCLUSIONS: Families are uncertain whether or how they can participate in the decision-making process. They need unambiguous communication and honest information to be able to take part in the decision-making process. We suggest that clinicians in Norwegian ICUs need more training in the knowledge and skills of effective communication with families of dying patients. PMID- 21626243 TI - Diagnostic reference levels for mammography in BreastScreen Queensland. AB - Diagnostic reference levels assist in the optimisation of radiation exposure parameters within a medical imaging facility. As no Australian DRLs currently exist, radiation doses from mammography in BreastScreen Queensland are analysed. Program-based DRLs of 1.1 and 1.4 mGy are proposed for digital radiography and computed radiography mammography systems, respectively. PMID- 21626245 TI - Choosing a teacher. PMID- 21626244 TI - HIV/AIDS prevention, faith, and spirituality among black/African American and Latino communities in the United States: strengthening scientific faith-based efforts to shift the course of the epidemic and reduce HIV-related health disparities. AB - Black/African American and Latino communities are disproportionately affected by the domestic HIV/AIDS epidemic. Blacks/African Americans and Latinos are also more likely to report a formal, religious, or faith affiliation when compared with non-Hispanic whites. As such, faith leaders and their institutions have been identified in the National HIV/AIDS Strategy as having a vital role to serve in reducing: (1) HIV-related health disparities and (2) the number of new HIV infections by promoting non-judgmental support for persons living with and at risk for HIV/AIDS and by serving as trusted information resources for their congregants and communities. We describe faith doctrines and faith-science partnerships that are increasing in support of faith-based HIV prevention and service delivery activities and discuss the vital role of these faith-based efforts in highly affected black/African American and Latino communities. PMID- 21626246 TI - For Jean with thanks. PMID- 21626248 TI - From a community mental-health center: Shared reflections. PMID- 21626247 TI - Dealing creatively with suffering - The living death. PMID- 21626249 TI - The problem-oriented record in pastoral counseling. PMID- 21626250 TI - The "three rs" of sex. AB - We have seen that sex has three functions, the reproductive, the relational, and the recreational. We have shown it probable that a great deal of unnecessary marital conflict comes from one party holding relational sex to be all-important while the other considers recreational sex to be of more importance. Finally, we have indicated that it is most likely that if they can compromise this difficulty by each meeting the other on his own ground a part of the time, much of the haggling will stop and many unnecessary divorces will be avoided. PMID- 21626251 TI - The cross and the seed: Active and receptive spiritualities. PMID- 21626252 TI - The rabbi and the psychiatrist: Effective counseling for youth. PMID- 21626253 TI - Jesus and assertiveness. PMID- 21626254 TI - Religious views of parents of retarded children. PMID- 21626256 TI - A systematic review and meta-analysis of applications of the Self-Report Habit Index to nutrition and physical activity behaviours. AB - BACKGROUND: Health behaviour models typically neglect habitual action. The Self Report Habit Index (SRHI) permits synthesis of evidence of the influence of habit on behaviour. PURPOSE: The purpose of this study is to review evidence around mean habit strength, habit-behaviour correlations, and habit x intention interactions, from applications of the SRHI to dietary, physical activity, and active travel behaviour. METHOD: Electronic database searches identified 126 potentially relevant papers. Twenty-two papers (21 datasets) passed eligibility screening. Mean scores and correlations were meta-analysed using fixed, random and mixed effects, and interactions were synthesised via narrative review. RESULTS: Twenty-three habit-behaviour correlations and nine habit x intention interaction tests were found. Typical habit strength was located around the SRHI midpoint. Weighted habit-behaviour effects were medium-to-strong (fixed: r + = 0.44; random: r + = 0.46). Eight tests found that habit moderated the intention behaviour relation. CONCLUSION: More comprehensive understanding of nutrition and activity behaviours will be achieved by accounting for habitual responses to contextual cues. PMID- 21626257 TI - Comparison of survival rate in primary non-small-cell lung cancer among elderly patients treated with radiofrequency ablation, surgery, or chemotherapy. AB - PURPOSE: We retrospectively compared the survival rate in patients with non-small cell lung cancer (NSCLC) treated with radiofrequency ablation (RFA), surgery, or chemotherapy according to lung cancer staging. MATERIALS AND METHODS: From 2000 to 2004, 77 NSCLC patients, all of whom had WHO performance status 0-2 and were >60 years old, were enrolled in a cancer registry and retrospectively evaluated. RFA was performed on patients who had medical contraindications to surgery/unsuitability for surgery, such as advanced lung cancer or refusal of surgery. In the RFA group, 40 patients with inoperable NSCLC underwent RFA under computed tomography (CT) guidance. These included 16 patients with stage I to II cancer and 24 patients with stage III to IV cancer who underwent RFA in an adjuvant setting. In the comparison group (n = 37), 13 patients with stage I to II cancer underwent surgery; 18 patients with stage III to IV cancer underwent chemotherapy; and 6 patients with stage III to IV cancer were not actively treated. The survival curves for RFA, surgery, and chemotherapy in these patients were calculated using Kaplan-Meier method. RESULTS: Median survival times for patients treated with (1) surgery alone and (2) RFA alone for stage I to II lung cancer were 33.8 and 28.2 months, respectively (P = 0.426). Median survival times for patients treated with (1) chemotherapy alone and (2) RFA with chemotherapy for stage III to IV cancer were 29 and 42 months, respectively (P = 0.03). CONCLUSION: RFA can be used as an alternative treatment to surgery for older NSCLC patients with stage I to II inoperable cancer and can play a role as adjuvant therapy with chemotherapy for patients with stage III to IV lung cancer. PMID- 21626258 TI - Percutaneous decompression of lumbar spinal stenosis with a new interspinous device. AB - OBJECTIVE: This study was designed to evaluate the feasibility of the implantation of a new interspinous device (Falena) in patients with lumbar spinal stenosis. The clinical outcomes and imaging results were assessed by orthostatic MR during an up to 6-month follow-up period. METHODS: Between October 2008 and February 2010, the Falena was implanted at a single level in 26 patients (17 men; mean age, 69 (range, 54-82) years) who were affected by degenerative lumbar spinal stenosis. All of the patients were clinically evaluated before the procedure and at 1 and 3 months. Furthermore, 20 patients have completed a 6 month follow-up. Pain was assessed before and after the intervention using the Visual Analogue Scale score and the Oswestry Disability Index questionnaire. Orthostatic MR imaging was performed before the implantation and at 3 months to assess the correlation with the clinical outcome. RESULTS: The mean ODI score decreased from 48.9 before the device implantation to 31.2 at 1 month (p < 0.0001). The mean VAS score decreased from 7.6 before to 3.9 (p < 0.0001) at 1 month and 3.6 at 3 months after the procedure (p = 0.0115). These values were stable at 6 months evaluation. No postimplantation major complications were recorded. MRI evaluation documented in most cases an increased size of the spinal canal area. Similarly a bilateral foraminal area improvement was found. The variation of the intervertebral space height measured on the posterior wall was not significant. CONCLUSIONS: In our preliminary experience with the Falena in a small cohort of patients, we obtained clinical and imaging results aligned to those reported with similar interspinous devices. PMID- 21626259 TI - Vibrational dynamics of bio- and nano-filaments in viscous solution subjected to ultrasound: implications for microtubules. AB - In this paper, using a new analytical method, we solved the beam equation for a uniform bio- and nano-filament in a viscous solution. The filament is assumed to be attached at its two ends and driven by ultrasound plane waves. To obtain analytical solutions, we converted the beam equation to an equation that allows us the use of the method of separation of variables. We then reconstructed the solution of the original beam equation from the solution of the converted equation. Subsequently, we have used the parametric equations derived in this paper to investigate the resonance condition for a microtubule (MT) in an aqueous solution. We show that by using ultrasound plane waves, one cannot satisfy a resonance condition for MTs treated as rigid rods. In order to achieve resonance, a single mode of the MT vibration must be excited with a harmonic number larger than a threshold value found here. Single mode excitation not only helps to transfer a minimum amount of energy to the surrounding medium compared with multi mode excitation, but it also allows for a simultaneous high amplitude and high mode quality that is impossible using plane waves. In order to overcome this difficulty, we propose to use an ultrasound generation device as a potential technical solution characterized by both frequency control and optimized energy transfer to the MT. Finally, the minimum required intensity of the ultrasound at the location of the MT in order to break it is shown to be on the order of 10(5) W/m(2), which corresponds to 170 dB. PMID- 21626260 TI - Characterization of spontaneous Parkinsonism in drug-naive patients with nonaffective psychotic disorders. AB - Spontaneous Parkinsonism (SP) in schizophrenia-related disorders is poorly characterized. The objective of this study was to examine the concordance and clinical validity of alternative definitions of SP in patients with nonaffective psychotic disorders. Two-hundred drug-naive patients with nonaffective psychotic disorders were examined for core parkinsonian signs, including bradykinesia, rigidity, and tremor, and diagnosed of SP according to the Simpson-Angus Scale (SAS) cutoff criterion, the UK Parkinson's disease brain bank (UKPDBB) criteria, the National Institute of Neurological Disorders and Stroke (NINDS) criteria, and criteria requiring the presence of all three core features (full syndrome criteria). Parkinsonian signs and criteria were examined in relation to a number of relevant clinical variables. The most frequent sign was rigidity (33.5%) followed by bradykinesia (16%) and tremor (12%). The prevalence rate of SP according to the SAS cutoff criterion, the UKPDBB criteria, the NINDS criteria for possible and probable SP, and the full syndrome criteria were 20.5, 13, 25.5, 18.5, and 4%, respectively. Bradykinesia was specifically related to negative symptoms, rigidity to neurological soft signs, and tremor to dyskinetic movements. The set of criteria showing more associations with clinical variables were the NINDS criteria for probable SP. Patients fulfilling these criteria had higher ratings for poor premorbid adjustment, negative symptoms, dyskinesia, neurological soft signs, and poor global treatment response than those without that diagnosis. The NINDS criteria for probable SP, i.e., presence of any two of the three core parkinsonian signs, seem to be the most suitable for clinical and research purposes. PMID- 21626261 TI - Anesthetic management of a patient with congenital diaphragmatic eventration. AB - Congenital diaphragmatic eventration is uncommon in adults and is caused by paralysis, aplasia or atrophy of the muscular fibers of the diaphragm. It may cause severe dyspnea, orthopnea and hypoxia in adult patients. Most symptomatic patients may be managed efficiently without the need for surgical correction, although any event that leads to an increase in intra-abdominal pressure puts them at the risk of spontaneous diaphragmatic rupture. This case report presents the successful anesthetic management of an adult female with congenital diaphragmatic eventration undergoing diagnostic laparoscopy and hysteroscopy using a total intravenous anesthesia technique. Essential steps to prevent any rise in intrathoracic and intra-abdominal pressures along with care to minimize intragastric volume were taken. PMID- 21626262 TI - In reply to comment on "Lidocaine spray 10 min prior to intubation: effects on postoperative sore throat". PMID- 21626263 TI - Anaphylaxis to pantoprazole during general anesthesia. AB - The most frequent causes of anaphylaxis during anesthesia are neuromuscular blocking agents, antibiotics, and latex. Proton pump inhibitors (PPI) are widely used during major surgery for the prevention of stress ulcers, but cases of perioperative anaphylactic reactions to these have rarely been reported. We present a 50-year-old male patient who experienced an episode of anaphylaxis with hypoxemia, hypotension, tachycardia, and generalized erythema after intravenous injection of pantoprazole 40 mg and methylprednisolone 1 g during general anesthesia. After resuscitation, the patient recovered without any sequelae. Six months after the surgery, a skin test was positive to pantoprazole. PMID- 21626264 TI - Overexpression of cinnamate 4-hydroxylase gene enhances biosynthesis of decursinol angelate in Angelica gigas hairy roots. AB - Angelica gigas is a medicinal plant that produces pyranocoumarins, including decursin (D) and decursinol angelate (DA), which have neuroprotective, anticancer, and antiandrogenic effects. In this study, the coumarin biosynthetic pathway was engineered to increase the production of DA. Specifically, a vector was constructed which contained the A. gigas phenylalanine ammonia-lyase (AgPAL) and cinnamate 4-hydroxylase (AgC4H) genes that were driven by the cauliflower mosaic virus (CaMV) 35S promoter. Transgenic hairy roots that overexpressed AgPAL or AgC4H genes were obtained by using an Agrobacterium rhizogenes-mediated transformation system. Among them, only AgC4H-transgenic hairy root lines produced more DA than control transgenic hairy root lines. The enhanced gene expression corresponded to elevated C4H activities. This study showed the importance of C4H in the production of DA in A. gigas hairy root culture. PMID- 21626265 TI - Patterns of information behavior and prostate cancer knowledge among African American men. AB - The purposes of this study are to explore cancer information acquisition patterns among African-American men and to evaluate relationships between information acquisition patterns and prostate cancer prevention and control knowledge. A random sample of 268 men participated in a statewide interviewer-administered, telephone survey. Men classified as non-seekers, non-medical source seekers, and medical source seekers of prostate cancer information differed on household income, level of education, and beliefs about personal risk for developing prostate cancer. Results from multiple regression analysis indicated that age, education, and information-seeking status were associated with overall levels of prostate cancer knowledge. Results from logistic regression analyses indicated that men who included physicians as one of many information resources (medical source seekers) had superior knowledge over non-seekers and non-medical source seekers on 33% of individual knowledge details. The findings emphasize the need to connect lower-income and lower-educated African-American men to physicians as a source of prostate cancer control information. PMID- 21626267 TI - APP and APLP1 are degraded through autophagy in response to proteasome inhibition in neuronal cells. AB - Amyloid beta (Abeta) precursor protein (APP) is a key protein in the pathogenesis of Alzheimer's disease (AD). Both APP and its paralogue APLP1 (amyloid beta precursor-like protein 1) have multiple functions in cell adhesion and proliferation. Previously it was thought that autophagy is a novel beta-amyloid peptide (Abeta)-generating pathway activated in AD. However, the protein proteolysis of APLP1 is still largely unknown. The present study shows that APLP1 is rapidly degraded in neuronal cells in response to stresses, such as proteasome inhibition. Activation of the endoplasmic reticulum (ER) stress by proteasome inhibitors induces autophagy, causing reduction of mature APLP1/APP. Blocking autophagy or JNK stress kinase rescues the protein expression for both APP and APLP1. Therefore, our results suggest that APP/APLP1 is degraded through autophagy and the APLP1 proteolysis is mainly mediated by autophagy-lysosome pathway. PMID- 21626268 TI - Retinoic acid inducible gene-I, more than a virus sensor. AB - Retinoic acid inducible gene-I (RIG-I) is a caspase recruitment domain (CARD) containing protein that acts as an intracellular RNA receptor and senses virus infection. After binding to double stranded RNA (dsRNA) or 5'-triphosphate single stranded RNA (ssRNA), RIG-I transforms into an open conformation, translocates onto mitochondria, and interacts with the downstream adaptor mitochondrial antiviral signaling (MAVS) to induce the production of type I interferon and inflammatory factors via IRF3/7 and NF-kappaB pathways, respectively. Recently, accumulating evidence suggests that RIG-I could function in non-viral systems and participate in a series of biological events, such as inflammation and inflammation related diseases, cell proliferation, apoptosis and even senescence. Here we review recent advances in antiviral study of RIG-I as well as the functions of RIG-I in other fields. PMID- 21626269 TI - Willingness to vaccinate or get vaccinated with an intradermal seasonal influenza vaccine: a survey of general practitioners and the general public in France and Germany. AB - INTRODUCTION: The elderly are at high risk of severe seasonal influenza and influenza-related death. Annual vaccination can effectively prevent influenza and its complications, and is recommended in the elderly. In the present study, surveys were undertaken in France and Germany to determine whether INTANZA (sanofi pasteur, Val-de-Reuil, France), the first intradermal influenza vaccine, administered using an innovative microneedle injection system, might influence physicians' likelihood of recommending influenza vaccination or the likelihood that the general public would seek influenza vaccination. METHODS: Physicians (France: n=260; Germany: n=223) and members of the general public aged >= 50 years (France: n=1706; Germany: n=1072) completed online surveys. Details of the INTANZA delivery system, and a "product profile" based on the properties of INTANZA, were presented. RESULTS: Most physicians and the general public found INTANZA and its microneedle injection system appealing. The main benefit of INTANZA, as perceived by physicians and the public, was the small needle size. Physicians also found the high immunogenicity compared with conventional intramuscular (IM) vaccines attractive. The majority of physicians believed that INTANZA would strongly help them to recommend vaccination to their unvaccinated patients (66% to 91%, depending upon patient characteristics); most (61% to 78%) would prefer to prescribe INTANZA rather than an IM vaccine. More than two-thirds of the unvaccinated general public would prefer INTANZA over IM vaccines, and the option of vaccination with INTANZA would encourage a large proportion of them to get vaccinated (60% to 74%), if it was recommended and they were given the choice. Physicians (>= 82%) agreed that INTANZA may help increase vaccination coverage rates. CONCLUSION: The results of these surveys indicate that the availability of INTANZA may encourage physicians to recommend influenza vaccination, and members of the general public to get vaccinated. INTANZA may help to improve seasonal influenza vaccination coverage rates. PMID- 21626270 TI - Visual discrimination, sequential learning and memory retrieval in the Australian desert ant Melophorus bagoti. AB - Bees, wasps and ants--so-called central-place foragers--need potent homing strategies to return to their nest. Path integration and view-based landmark guidance are the key strategies for the ants' navigation. For instance, they memorise different views in a sequence (sequential memory) but also have a step counter that informs them about the covered distance during each foraging trip (odometer). The sequential memory and the odometer information can act as contextual cues during travel for retrieving the appropriate stored view. When and which cue is used at different stages and lengths of the foraging trips is still unknown. In this study, we examined how the Australian desert ant Melophorus bagoti uses sequential memory and odometric information to retrieve visual memories. Using a set-up made out of channels and two-choice boxes (Y mazes), we demonstrate first that M. bagoti foragers are able to learn and discriminate a variety of visual stimuli in a sequence of views along the inbound trip back to the nest. We then forced the homing ants to encounter a fixed sequence of two visual patterns during their inbound trips. By manipulating the position and distance of the visual stimuli and decision boxes, we could set the two contextual cues (sequential memory and odometer) into conflict. After the short 4-m outbound distance, a preference for odometric information as a contextual cue was found, but after the long 8-m outbound distance, ants relied primarily on their sequential memory retrieval. Odometer precision deteriorates with increasing travel distance, and accordingly, our findings imply that desert ants may be relying on the most reliable contextual cue for retrieving visual memories. PMID- 21626272 TI - Histochemical localization of caldesmon isoforms in colon adenocarcinoma and lymph node metastases. AB - Alternative splicing of the caldesmon gene results in high (h-caldesmon) and low (l-caldesmon) molecular weight isoforms of the cytoskeleton-associated protein caldesmon. h-Caldesmon is predominantly expressed not only in smooth-muscle cells but also in pericryptal fibroblasts in colon. l-Caldesmon is widely expressed and localized in podosomes/invadopodia. Studies with transformed and cancer cell lines suggest that a reduction in l-caldesmon facilitates podosome/invadopodia formation and metastasis. We investigated caldesmon isoforms in colon adenocarcinoma and their lymph node metastases using immunohistochemistry. Caldesmon immunoreactivity of colon adenocarcinoma primary tumors and lymph node metastases was very similar. l-Caldesmon immunoreactivity of cancer epithelial cells in primary tumors and lymph node metastases varied ranging from reduced to stronger as compared to immunoreactivity of normal areas. The variation did not show a consistent relation to the tumor center or invasive margin. While h caldesmon immunoreactivity of pericryptal fibroblasts and blood vessels in the stroma of primary tumors and lymph node metastases was markedly reduced, cancer associated fibroblasts and blood vessels in the stroma were strongly immunoreactive for l-caldesmon. Our results show a differential behavior of h- and l-caldesmon isoforms in epithelium and stroma of colon adenocarcinoma and lymph node metastases. PMID- 21626271 TI - Experimental approaches to study functional recovery following cerebral ischemia. AB - Valid experimental models and behavioral tests are indispensable for the development of therapies for stroke. The translational failure with neuroprotective drugs has forced us to look for alternative approaches. Restorative therapies aiming to facilitate the recovery process by pharmacotherapy or cell-based therapy have emerged as promising options. Here we describe the most common stroke models used in cell-based therapy studies with particular emphasis on their inherent complications, which may affect behavioral outcome. Loss of body weight, stress, hyperthermia, immunodepression, and infections particularly after severe transient middle cerebral artery occlusion (filament model) are recognized as possible confounders to impair performance in certain behavioral tasks and bias the treatment effects. Inherent limitations of stroke models should be carefully considered when planning experiments to ensure translation of behavioral data to the clinic. PMID- 21626273 TI - The oblique occipital sinus: a review of anatomy and imaging characteristics. AB - Variations of the dural venous sinuses may result in inaccurate imaging interpretation or complications during surgical approaches. One variation of the dural venous sinuses reported infrequently in the literature is the oblique occipital sinus. The present paper reviews this anatomy and offers illustrations of the cadaveric and imaging findings seen with this venous variation. PMID- 21626274 TI - Intra-articular ossicle in interphalangeal joint of the great toe and clinical implication. AB - Failure of closed reduction for an interphalangeal joint dislocation of the great toe resulted from an intra-articular ossicle interposed between the articular cartilages of the phalanges. The knowledge concerning the intra-articular ossicle is unclear. A study was thus carried out on 100 fresh great toes to document the appearance, number, size, and location of the intra-articular ossicle found in the interphalangeal joint of the great toe. Roentgenographic studies of the joint revealed 86% of bony mass representing either the sesamoid bone or the intra articular ossicle. Anatomical studies revealed no sesamoid bone in the flexor hallucis longus tendon. There was 88% of intra-articular ossicle on the dorsal surface of the plantar capsule of the interphalangeal joint. A medial surgical approach to reduce the irreducible dislocation is thus suggested as easier and safer than other approaches. PMID- 21626275 TI - Anatomic study of fabella and its surrounding structures in a Chinese population. AB - PURPOSE: To explore the anatomic features of the fabella and its relationship with the common peroneal nerve and the fabellofibular ligament, so as to provide anatomical evidence for clinical diagnosis and treatment of fabella diseases in a Chinese population. METHODS: Sixty-one formalin-fixed knee specimens were obtained for anatomic dissection. Structural features of the fabella were investigated by radiological and histological tests. RESULTS: There were 53 cases (86.89%) with fabellae in the lateral head of the gastrocnemius muscle, including 34 bony ones (55.74%), whereas only 6 cases had fabellae in the medial head (9.84%). The fabellae were accompanied by common peroneal nerves on their surfaces in 11 cases (20.8%), and the presence of the fabella was not generally predictive of a fabellofibular ligament. As much as 57.9% of the cartilage fabellae were not visualized on radiograph. The structure of the ossified fabella is similar to a typical long bone. CONCLUSIONS: Fabellae were mainly present in the lateral head of the gastrocnemius muscle in a large proportion of the Chinese population. More than half of the cartilage fabellae were not visualized on radiograph. Its clinical significance could not be ignored by physicians and anatomists. PMID- 21626276 TI - Bilateral malrotation and lobulation of kidney with altered hilar anatomy: a rare congenital variation. AB - Variations of the structure and position of the kidney along with the variations of renal vessels are the most frequently reported. Rotational variations form a rare entity that is not cited in most of the embryology textbooks. During educational cadaveric dissection of a 57-year-old female, a complex picture of bilateral anatomical variants was encountered. Bilateral malrotation of kidney, bilateral lobulated kidneys along with open hilum of kidney was observed, so that the contents of the sinus were exposed. The renal pelvis was present anterior to the renal vessels instead of posterior position.The right kidney in addition showed lower lumbar position with three supplementary arteries and two veins. The right ovarian vein arched over the laterally rotated hilum of kidney and drained into superior renal vein instead of inferior vena cava. This is a rare case in which such numbers of congenital variations are seen. These variations of kidney have embryological base. Rotational variation though comparatively rare assumes great importance in view of the present day surgical procedures like percutaneous nephrectomy, renal transplantation, etc. PMID- 21626277 TI - Comparison of stability in the operative treatment of pelvic injuries in a finite element model. AB - PURPOSE OF THE STUDY: The comparison of the stability of four surgical methods for the treatment of vertically and rotationally unstable type C pelvic ring injuries. METHODS: We produced a type C pelvic ring injury (type Denis II fracture of the sacrum and symphysiolysis) on a finite element model, in the case of standing on both feet. We stabilized the symphysiolysis with a five-hole reconstruction plate; the sacrum fracture was fixed in the first experiment with two, two-hole reconstruction plates on the ventral surface, in the second one we applied dorsally the transsacral, narrow DC plate, in the third one with KFI-H plate, and in the last one with iliosacral screw. Finite element modeling was performed by the use of the ALGOR software. Not only bones and joints, but joints and mechanically important ligaments were modeled as well. We measured the shift between the two surfaces of the fracture gap, compared to the results of measurements accomplished on cadaver models. RESULTS: Larger shift could be elicited after transsacral plating than after direct plating. These results correspond to those of the parallel investigation of the bony ligamentous cadaver pelvis specimens. The shift values after KFI-H plating and iliosacral screw fixation are larger than after direct plating, but smaller than after transsacral plating. The tension created in the implants is less than the allowed values; therefore, the choice of operation should depend on the type of injury. CONCLUSIONS: The finite element model may be utilized for the comparison of different methods of osteosynthesis for the treatment of injuries described above. Due to several difficulties in investigations performed on cadaver specimens, this model has undoubted utility. PMID- 21626278 TI - Phase-based arterial input functions in humans applied to dynamic contrast enhanced MRI: potential usefulness and limitations. AB - OBJECT: Phase-based arterial input functions (AIFs) provide a promising alternative to standard magnitude-based AIFs, for example, because inflow effects are avoided. The usefulness of phase-based AIFs in clinical dynamic contrast enhanced MRI (DCE-MRI) was investigated, and relevant pitfalls and sources of uncertainty were identified. MATERIALS AND METHODS: AIFs were registered from eight human subjects on, in total, 21 occasions. AIF quality was evaluated by comparing AIFs from right and left internal carotid arteries and by assessing the reliability of blood plasma volume estimates. RESULTS: Phase-based AIFs yielded an average bolus peak of 3.9 mM and a residual concentration of 0.37 mM after 3 min, (0.033 mmol/kg contrast agent injection). The average blood plasma volume was 2.7% when using the AIF peak in the estimation, but was significantly different (p < 0.0001) and less physiologically reasonable when based on the AIF tail concentration. Motion-induced phase shifts and accumulation of contrast agent in background tissue regions were identified as main sources of uncertainty. CONCLUSION: Phase-based AIFs are a feasible alternative to magnitude AIFs, but sources of errors exist, making quantification difficult, especially of the AIF tail. Improvement of the technique is feasible and also required for the phase-based AIF approach to reach its full potential. PMID- 21626279 TI - Rescuing of deficient killing and phagocytic activities of macrophages derived from non-obese diabetic mice by treatment with geldanamycin or heat shock: potential clinical implications. AB - Diabetes mellitus type 1 (DMT1) is an autoimmune disease characterized by the destruction of insulin-producing cells in the pancreas. Diabetic patients are more susceptible to recurrent and uncontrolled infections, with worse prognoses than in healthy individuals. Macrophages (MPhis) derived from DMT1 individuals have compromised mounting of inflammatory and immune responses. The mechanisms responsible for these alterations remain unknown. It has been shown that the presence of extra- and intracellular heat shock proteins (hsp) positively modulates immune cell function. Using naive MPhis derived from non-obese diabetic (NOD) mice, a well-established mouse model for DMT1, we demonstrate that heat shock (HS) as well as treatment with geldanamycin (GA), significantly improves diabetic MPhi activation, resulting in increased phagocytosis and killing of bacteria. Induction of HS did not affect the aberrant NOD-MPhi cytokine profile, which is characterized by elevated IL-10 levels and normal tumor necrosis factor alpha. Our observations were consistent at pre-diabetic (normal random blood glucose) and diabetic (random blood glucose greater than 250 mg/dl) stages, suggesting that HS and GA treatment may compensate for intrinsic genetic alterations present in diabetic cells regardless of the stage of the disease. The mechanisms associated to this phenomenon are unknown, but they may likely be associated with the induction of hsp expression, a common factor between HS and GA treatment. Our results may open a new field for non-classical function of hsp and indicate that hsp expression may be used as a part of therapeutic approaches for the treatment of complications associated with DMT1 as well as other autoimmune diseases. PMID- 21626280 TI - TM4SF1: a tetraspanin-like protein necessary for nanopodia formation and endothelial cell migration. AB - Transmembrane-4-L-six-family-1 (TM4SF1) is a tetraspanin-like membrane protein that is highly and selectively expressed by cultured endothelial cells (EC) and, in vivo, by EC lining angiogenic tumor blood vessels. TM4SF1 is necessary for the formation of unusually long (up to a 50 MUm), thin (~100-300 nm wide), F-actin poor EC cell projections that we term 'nanopodia'. Immunostaining of nanopodia at both the light and electron microsopic levels localized TM4SF1 in a regularly spaced, banded pattern, forming TM4FS1-enriched domains. Live cell imaging of GFP transduced HUVEC demonstrated that EC project nanopodia as they migrate and interact with neighboring cells. When TM4SF1 mRNA levels in EC were increased from the normal ~90 mRNA copies/cell to ~400 copies/cell through adenoviral transduction, EC projected more and longer nanopodia from the entire cell circumference but were unable to polarize or migrate effectively. When fibroblasts, which normally express TM4SF1 at ~5 copies/cell, were transduced to express TM4SF1 at EC-like levels, they formed typical TM4SF1-banded nanopodia, and broadened, EC-like lamellipodia. Mass-spectrometry demonstrated that TM4SF1 interacted with myosin-10 and beta-actin, proteins involved in filopodia formation and cell migration. In summary, TM4SF1, like genuine tetraspanins, serves as a molecular organizer that interacts with membrane and cytoskeleton associated proteins and uniquely initiates the formation of nanopodia and facilitates cell polarization and migration. PMID- 21626281 TI - A simple bias correction in linear regression for quantitative trait association under two-tail extreme selection. AB - Selective genotyping can increase power in quantitative trait association. One example of selective genotyping is two-tail extreme selection, but simple linear regression analysis gives a biased genetic effect estimate. Here, we present a simple correction for the bias. PMID- 21626282 TI - Simplified MPN method for enumeration of soil naphthalene degraders using gaseous substrate. AB - We describe a simplified microplate most-probable-number (MPN) procedure to quantify the bacterial naphthalene degrader population in soil samples. In this method, the sole substrate naphthalene is dosed passively via gaseous phase to liquid medium and the detection of growth is based on the automated measurement of turbidity using an absorbance reader. The performance of the new method was evaluated by comparison with a recently introduced method in which the substrate is dissolved in inert silicone oil and added individually to each well, and the results are scored visually using a respiration indicator dye. Oil-contaminated industrial soil showed slightly but significantly higher MPN estimate with our method than with the reference method. This suggests that gaseous naphthalene was dissolved in an adequate concentration to support the growth of naphthalene degraders without being too toxic. The dosing of substrate via gaseous phase notably reduced the work load and risk of contamination. The result scoring by absorbance measurement was objective and more reliable than measurement with indicator dye, and it also enabled further analysis of cultures. Several bacterial genera were identified by cloning and sequencing of 16S rRNA genes from the MPN wells incubated in the presence of gaseous naphthalene. In addition, the applicability of the simplified MPN method was demonstrated by a significant positive correlation between the level of oil contamination and the number of naphthalene degraders detected in soil. PMID- 21626283 TI - Optimizing culture conditions of a porcine epithelial cell line IPEC-J2 through a histological and physiological characterization. AB - The high similarity between pigs and humans makes pigs a good gastrointestinal (GI) model for humans. Recently an epithelial cell line originating from the jejunum of pig (IPEC-J2) became available. Once validated, this model can be used to investigate the complex interactions occurring in the intestine. The advantages of using IPEC-J2 as in vitro model of the GI tract are the high resemblance between humans and pigs, and the ease of extrapolating in vitro to in vivo characteristics. In this study, the IPEC-J2 cells were functionally characterized by measuring the trans-epithelial electrical resistance (TEER), and by histological and ultrastructural studies. IPEC-J2 cells grown on six different permeable support systems, were investigated. The Transwell((r))-COL collagen coated membrane (1.12 cm(2)) showed the best results concerning time efficiency and TEER values. The optimum seeding density of 12 * 10(5) cells/mL ensured that after 9 days of differentiation a confluent monolayer was formed. The decrease in TEER values after a maximum had been reached, coincided with the ultrastructural development of apical microvilli. We conclude that IPEC-J2 cells grown on collagen-coated membranes represent a valuable in vitro model system for the small intestinal epithelium which can be of great interest for intestinal research. PMID- 21626284 TI - Desmopressin duration of antidiuretic action in patients with central diabetes insipidus. AB - The key question answered by this study is whether it is possible to deliver a pharmacokinetic and pharmacodynamic duration of antidiuretic action long enough to ensure adequate antidiuresis with two daily administrations of desmopressin in patients with central diabetes insipidus (CDI). We studied the efficacy and safety of desmopressin i.v. in 13 CDI patients using two 3-way crossover designs, in the doses 30, 60, 125 ng, and 125, 250 and 500 ng. Duration of action, minimum output rate, max osmolality and average osmolality during action (AUC osmolality) were measured every 30 min for the first 2 h during the infusion, and then every hour or every second hour until the urine output rate was greater than 2 ml/kg/30 min. The duration of antidiuretic action was 4, 8 and 11 h, respectively, for 125, 250, and 500 ng, increasing from 250 to 500 ng but for the remaining secondary dynamic efficacy parameters no difference could be detected based on descriptive statistics between the doses 250 and 500 ng, indicating that the upper plateau region of the dose-response curve had been reached. All treatment emergent adverse events were classified as unrelated or unlikely related to trial medication. No serious adverse events occurred. Data on duration of action indicates that it is possible to achieve antidiuretic control with 500 ng i.v. corresponding to 160 MUg orodispersible tablets twice daily in CDI patients. Today, the Minirin Melt label recommends the majority of CDI patients a dose of 60 to 120 MUg t.i.d. PMID- 21626285 TI - Unraveling the complexities of cannabinoid receptor 2 (CB2) immune regulation in health and disease. AB - It has become clear that the endocannabinoid system is a potent regulator of immune responses, with the cannabinoid receptor 2 (CB2) as the key component due to its high expression by all immune subtypes. CB2 has been shown to regulate immunity by a number of mechanisms including development, migration, proliferation, and effector functions. In addition, CB2 has been shown to modulate the function of all immune cell types examined to date. CB2 is a G(i) protein-coupled receptor and thus exhibits a complex pharmacology allowing both stimulatory and inhibitory signaling that depends on receptor expression levels, ligand concentration, and cell lineage specificities. Here, we discuss both in vitro and in vivo experimental evidence that CB2 is a potent regulator of immune responses making it a prime target for the treatment of inflammatory diseases. PMID- 21626286 TI - Is mandatory screening for obstructive sleep apnea with polysomnography in all severely obese patients indicated? AB - PURPOSE: The study aims to assess the risk factors for the presence and severity of obstructive sleep apnea (OSA) among severely obese patients evaluated for bariatric surgery. PATIENTS AND METHODS: Polysomnography recordings were performed in consecutive patients undergoing Roux-en-Y gastric bypass from January 2004 to January 2007. Sleep apnea was noted as present or absent and graded from mild to severe according to the apnea/hypopnea index. Patient gender, age, weight, height, body mass index, neck circumference, and waist circumference were recorded. RESULTS: A total of 132 patients were included in the study group, and 85 patients had a confirmed diagnosis of OSA (64.4%). The prevalence of OSA was 55.7% in female and 77.4% in male. The prevalence of moderate or severe sleep apnea was higher in males (71.6%) than in females (31.6%). In OSA patients, body mass index (p = 0.020), neck circumference (p < 0.001), and age (p = 0.003) were higher as compared with obese patients without OSA, whereas no differences were found in waist circumference between groups. After multiple regression analysis, body mass index, age, and male gender were independent predictors of sleep apnea. In the female group, age greater than 49 years was the only significant predictor of moderate or severe OSA (odds ratio 5.42 (95% confidence interval 1.61-18.1); p = 0.006). CONCLUSION: Males and females with age greater than 49 years are at greatest risk for OSA. Preoperative sleep studies should be mandatory in this group of severely obese patients. PMID- 21626287 TI - Alleviation of exogenous oligochitosan on wheat seedlings growth under salt stress. AB - Hydroponic experiments were carried out to study the role of oligochitosan in enhancing wheat (Triticum aestivum L.) resistance to salt stress. Data were collected on plant biomass, chlorophyll content, photosynthetic rate (P (n)), stomatal conductance (g (s)), proline content, antioxidant enzyme activities, and malondialdehyde (MDA) content. Under 150 mM salt stress, plant growth was significantly inhibited. Shoot length, root length, and dry weight were sharply reduced by 26%, 31%, and 20%, respectively, of the control. Superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) were increased by 28%, 13%, and 26%, respectively, of the control and MDA content largely accumulated, which was 1.5 fold of the control. However, 0.0625% oligochitosan pretreatment alleviated the adverse effects of salt stress, which was reflected by increasing root length, shoot length, dry weight, chlorophyll content, P (n,) and g (s). Furthermore, it also showed that oligochitosan pretreatment significantly increased antioxidant enzyme (SOD, CAT and POD) activities, and reduced MDA content in leaves. Meanwhile, the accumulation of proline was markedly accelerated. The results indicated that oligochitosan pretreatment ameliorated the adverse effects and partially protected the seedlings from salt stress during the following growth period. PMID- 21626288 TI - A classification study of human beta3-adrenergic receptor agonists using BCUT descriptors. AB - Experimental EC(50)s for 202 human beta(3)-AR agonists are used to develop classification models as a potential screening tool for a large library of target compounds before synthesis. A variable selection approach from random forests (VS RF) is used to extract the structural information most relevant to the human beta(3)-AR activation properties of the collected data set. The obtained results indicate that the VS-RF method can be used for variable selection with smallest sets of non-redundant descriptors with highly predictive accuracy (Q (ex)% = 96% for the external prediction set). Thus, the proposed VS-RF models should be helpful for screening of potential human beta(3)-AR agonists before chemical synthesis in drug development. PMID- 21626290 TI - Endogenous tissue engineering: PTH therapy for skeletal repair. AB - Based on its proven anabolic effects on bone in osteoporosis patients, recombinant parathyroid hormone (PTH(1-34)) has been evaluated as a potential therapy for skeletal repair. In animals, the effect of PTH(1-34) has been investigated in various skeletal repair models such as fractures, allografting, spinal arthrodesis and distraction osteogenesis. These studies have demonstrated that intermittent PTH(1-34) treatment enhances and accelerates the skeletal repair process via a number of mechanisms, which include effects on mesenchymal stem cells, angiogenesis, chondrogenesis, bone formation and resorption. Furthermore, PTH(1-34) has been shown to enhance bone repair in challenged animal models of aging, inflammatory arthritis and glucocorticoid-induced bone loss. This pre-clinical success has led to off-label clinical use and a number of case reports documenting PTH(1-34) treatment of delayed-unions and non-unions have been published. Although a recently completed phase 2 clinical trial of PTH(1-34) treatment of patients with radius fracture has failed to achieve its primary outcome, largely because of effective healing in the placebo group, several secondary outcomes are statistically significant, highlighting important issues concerning the appropriate patient population for PTH(1-34) therapy in skeletal repair. Here, we review our current knowledge of the effects of PTH(1-34) therapy for bone healing, enumerate several critical unresolved issues (e.g., appropriate dosing regimen and indications) and discuss the long-term potential of this drug as an adjuvant for endogenous tissue engineering. PMID- 21626289 TI - Transforming growth factor-beta and atherosclerosis: interwoven atherogenic and atheroprotective aspects. AB - Age-related progression of cardiovascular disease is by far the largest health problem in the US and involves vascular damage, progressive vascular fibrosis and the accumulation of lipid-rich atherosclerotic lesions. Advanced lesions can restrict flow to key organs and can trigger occlusive thrombosis resulting in a stroke or myocardial infarction. Transforming growth factor-beta (TGF-beta) is a major orchestrator of the fibroproliferative response to tissue damage. In the early stages of repair, TGF-beta is released from platelets and activated from matrix reservoirs; it then stimulates the chemotaxis of repair cells, modulates immunity and inflammation and induces matrix production. At later stages, it negatively regulates fibrosis through its strong antiproliferative and apoptotic effects on fibrotic cells. In advanced lesions, TGF-beta might be important in arterial calcification, commonly referred to as "hardening of the arteries". Because TGF-beta can signal through multiple pathways, namely the SMADs, a MAPK pathway and the Rho/ROCK pathways, selective defects in TGF-beta signaling can disrupt otherwise coordinated pathways of tissue regeneration. TGF-beta is known to control cell proliferation, cell migration, matrix synthesis, wound contraction, calcification and the immune response, all being major components of the atherosclerotic process. However, many of the effects of TGF-beta are essential to normal tissue repair and thus, TGF-beta is often thought to be "atheroprotective". The present review attempts to parse systematically the known effects of TGF-beta on both the major risk factors for atherosclerosis and to isolate the role of TGF-beta in the many component pathways involved in atherogenesis. PMID- 21626291 TI - Smad phosphoisoform signals in acute and chronic liver injury: similarities and differences between epithelial and mesenchymal cells. AB - Hepatocellular carcinoma (HCC) usually arises from hepatic fibrosis caused by chronic inflammation. In chronic liver damage, hepatic stellate cells undergo progressive activation to myofibroblasts (MFB), which are important extracellular matrix-producing mesenchymal cells. Concomitantly, perturbation of transforming growth factor (TGF)-beta signaling by pro-inflammatory cytokines in the epithelial cells of the liver (hepatocytes) promotes both fibrogenesis and carcinogenesis (fibro-carcinogenesis). Insights into fibro-carcinogenic effects on chronically damaged hepatocytes have come from recent detailed analyses of the TGF-beta signaling process. Smad proteins, which convey signals from TGF-beta receptors to the nucleus, have intermediate linker regions between conserved Mad homology (MH) 1 and MH2 domains. TGF-beta type I receptor and pro-inflammatory cytokine-activated kinases differentially phosphorylate Smad2 and Smad3 to create phosphoisoforms phosphorylated at the COOH-terminal, linker, or both (L/C) regions. After acute liver injury, TGF-beta-mediated pSmad3C signaling terminates hepatocytic proliferation induced by the pro-inflammatory cytokine-mediated mitogenic pSmad3L pathway; TGF-beta and pro-inflammatory cytokines synergistically enhance collagen synthesis by activated hepatic stellate cells via pSmad2L/C and pSmad3L/C pathways. During chronic liver disease progression, pre-neoplastic hepatocytes persistently affected by TGF-beta together with pro inflammatory cytokines come to exhibit the same carcinogenic (mitogenic) pSmad3L and fibrogenic pSmad2L/C signaling as do MFB, thereby accelerating liver fibrosis while increasing risk of HCC. This review of Smad phosphoisoform-mediated signals examines similarities and differences between epithelial and mesenchymal cells in acute and chronic liver injuries and considers Smad linker phosphorylation as a potential target for the chemoprevention of fibro-carcinogenesis. PMID- 21626292 TI - Decreased NKG2D expression on NK cells correlates with impaired NK cell function in patients with gastric cancer. AB - BACKGROUND: Although malignant diseases are known to be associated with immune suppression, the detailed mechanisms involved are still unknown. NKG2D is an activating cell surface receptor expressed by natural killer (NK) cells and CD8+ T cells, and the engagement of NKG2D is extremely important for NK cell activation. Although decreased NKG2D expression on NK cells is closely related to immune evasion by some cancers, the immunopathological importance of this phenomenon in gastric cancer patients remains unclear. METHODS: NKG2D expression on NK cells was determined, using multicolor flow cytometry, to investigate the mechanisms responsible for immune evasion in gastric cancer patients. RESULTS: NKG2D expression on NK cells from gastric cancer patients was significantly lower than that in healthy controls. Also, NKG2D expression in advanced gastric cancer was significantly lower than that in early gastric cancer. NK cells from patients with lymph node metastasis expressed significantly lower levels of NKG2D than the NK cells from those without lymph node metastasis, and NKG2D expression on NK cells in gastric cancer tissue was significantly lower than that of circulating NK cells. NKG2D expression on NK cells obtained from cancer patients was restored after 48 h in culture with RPMI containing 10% AB serum. Furthermore, NKG2D expression on NK cells obtained after surgery was significantly higher than that before surgery. CONCLUSIONS: Decreased NKG2D expression on NK cells may be one of the key mechanisms responsible for NK cell dysfunction in gastric cancer. PMID- 21626293 TI - Early warning system for financially distressed hospitals via data mining application. AB - The aim of this study is to develop a Financial Early Warning System (FEWS) for hospitals by using data mining. A data mining method, Chi-Square Automatic Interaction Detector (CHAID) decision tree algorithm, was used in the study for financial profiling and developing FEWS. The study was conducted in Turkish Ministry of Health's public hospitals which were in financial distress and in need of urgent solutions for financial issues. 839 hospitals were covered and financial data of the year 2008 was obtained from Ministry of Health. As a result of the study, it was determined that 28 hospitals (3.34%) had good financial performance, and 811 hospitals (96.66%) had poor financial performance. According to FEWS, the covered hospitals were categorized into 11 different financial risk profiles, and it was found that 6 variables affected financial risk of hospitals. According to the profiles of hospitals in financial distress, one early warning signal was detected and financial road map was developed for risk mitigation. PMID- 21626294 TI - Deriving the operational procedure for the Universal Thermal Climate Index (UTCI). AB - The Universal Thermal Climate Index (UTCI) aimed for a one-dimensional quantity adequately reflecting the human physiological reaction to the multi-dimensionally defined actual outdoor thermal environment. The human reaction was simulated by the UTCI-Fiala multi-node model of human thermoregulation, which was integrated with an adaptive clothing model. Following the concept of an equivalent temperature, UTCI for a given combination of wind speed, radiation, humidity and air temperature was defined as the air temperature of the reference environment, which according to the model produces an equivalent dynamic physiological response. Operationalising this concept involved (1) the definition of a reference environment with 50% relative humidity (but vapour pressure capped at 20 hPa), with calm air and radiant temperature equalling air temperature and (2) the development of a one-dimensional representation of the multivariate model output at different exposure times. The latter was achieved by principal component analyses showing that the linear combination of 7 parameters of thermophysiological strain (core, mean and facial skin temperatures, sweat production, skin wettedness, skin blood flow, shivering) after 30 and 120 min exposure time accounted for two-thirds of the total variation in the multi dimensional dynamic physiological response. The operational procedure was completed by a scale categorising UTCI equivalent temperature values in terms of thermal stress, and by providing simplified routines for fast but sufficiently accurate calculation, which included look-up tables of pre-calculated UTCI values for a grid of all relevant combinations of climate parameters and polynomial regression equations predicting UTCI over the same grid. The analyses of the sensitivity of UTCI to humidity, radiation and wind speed showed plausible reactions in the heat as well as in the cold, and indicate that UTCI may in this regard be universally useable in the major areas of research and application in human biometeorology. PMID- 21626295 TI - Both gas chromatography and an electronic nose reflect chemical polymorphism of juniper shrubs browsed or avoided by sheep. AB - Chemical polymorphism may contribute to variation in browsing damage by mammalian herbivores. Earlier, we demonstrated that essential oil concentration in juniper, Juniperus communis, was negatively associated with herbivore browsing. The aim of the present study was to characterize the volatile chemical composition of browsed and non-browsed J. communis. By using either gas chromatography with flame ionization detection (GC-FID) or an electronic nose device, we could separate sheep-browsed or non-browsed juniper shrubs by their essential oil pattern and complex odor matrix. The main components of the essential oil from J. communis were monoterpenes. We distinguished three chemotypes, dominated either by alpha-pinene, sabinene, or delta-3-carene. Shrubs belonging to the alpha pinene- or sabinene-dominated groups were browsed, whereas all individuals with the delta-3-carene chemotype were unused by the local herbivores. The electronic nose also separated the browsed and non-browsed shrubs indicating that their odor matrix could guide sheep browsing. Responses of sheep could integrate the post ingestive effects of plant secondary metabolites with sensory experience that stems from odor-phytotoxin interactions. Chemotype diversity could increase the survival rate in the present population of J. communis as certain shrubs could benefit from relatively better chemical protection against the herbivores. PMID- 21626296 TI - Probiotics and dietary counselling targeting maternal dietary fat intake modifies breast milk fatty acids and cytokines. AB - PURPOSE: Breast milk fatty acids possess immunomodulatory properties, and new intervention strategies beyond supplementation of maternal diet with single oils are called for. The objective of the present study was to evaluate the effect of dietary intervention during pregnancy and breastfeeding on breast milk fatty acid and cytokine composition. METHODS: Pregnant women were randomised into three study groups: dietary intervention with probiotics (diet/probiotic) or with placebo (diet/placebo) and a control group (control/placebo). Dietary intervention included dietary counselling and provision of rapeseed oil-based food products. The probiotics used were Lactobacillus rhamnosus GG and Bifidobacterium lactis Bb12 in combination. Dietary intake was evaluated by food records at every trimester of pregnancy and 1 month postpartum. Breast milk samples were collected after birth (colostrum) and 1 month after delivery for fatty acid and cytokine analysis (n = 125). RESULTS: Dietary intervention improved the quality of fat in the diet. In breast milk, the proportion of alpha linolenic acid and total n-3 fatty acids was higher in both dietary intervention groups compared with control group (p < 0.05). In the diet/probiotic group, the gamma-linolenic acid content was higher compared with the diet/placebo group (p < 0.05). The concentrations of TNF-alpha, IL-10, IL-4 and IL-2 were higher in both dietary intervention groups compared with controls, and furthermore, long-chain n 3 fatty acids were associated with several cytokines in colostrum samples. CONCLUSION: The present intervention demonstrated the possibility of modifying breast milk immunomodulatory factors by dietary means. PMID- 21626297 TI - Acculturation and cardiovascular behaviors among Latinos in California by country/region of origin. AB - Despite generally lower socioeconomic status and worse access to healthcare, Latinos have better overall health outcomes and longer life expectancy than non Latino Whites. This "Latino Health Paradox" has been partially attributed to healthier cardiovascular (CV) behaviors among Latinos. However, as Latinos become more acculturated, differences in some CV behaviors disappear. This study aimed to explore how associations between acculturation and CV behaviors among Latinos vary by country of origin. Combined weighted data from the 2005 and 2007 California Health Interview Survey (CHIS) were used to investigate associations between acculturation level and CV behaviors among Latinos by country of origin. Among all Latinos, increased acculturation was associated with more smoking, increased leisure-time physical activity, and greater consumption of fast foods, but no change in fruit/vegetable and less soda intake. These trends varied, however, by Latino sub-groups from different countries of origin. Country of origin appears to impact associations between acculturation and CV behaviors among Latinos in complex ways. PMID- 21626298 TI - Racial/ethnic differences in hormonally-active hair product use: a plausible risk factor for health disparities. AB - Estrogen and endocrine-disrupting chemicals (EDCs) that are associated with several health outcomes have been found in hair products. We evaluated the proportion, frequency, duration, and content of hair products in a racially/ethnically diverse population. We recruited n = 301 African-American, African-Caribbean, Hispanic, and white women from the New York metropolitan area. We collected data on hair oil, lotion, leave-in conditioner, root stimulator, perm, and other product use. Estrogen and EDC information was collected from commonly used hair products' labels (used by >3% of population). African-American and African-Caribbean women were more likely to use all types of hair products compared to white women (P < 0.0001). Among hair product users, frequency varied significantly by race/ethnicity, but not duration. More African-Americans (49.4%) and African-Caribbeans (26.4%) used products containing placenta or EDCs compared to whites (7.7%). African-American and African-Caribbean women were more likely to be exposed to hormonally-active chemicals in hair products. PMID- 21626299 TI - Analysis of DNA methylation in human BK virus. AB - Human BK virus may cause nephropathy due to viral replication in patients who have undergone renal transplantation. However, the mechanism regulating replication of BKV is still not clear. Previous studies have suggested that epigenetic modifications may play a crucial role in virus replication. In this study, the DNA methylation profiles of five CpG sites located within the promoter/enhancer regions and nine CpG sites located within the early and late coding regions of the replicating BKV genome were investigated. BKV genomic DNA from mature virions and from the early and late phases of replicating BKV were examined for DNA methylation by bisulfite sequencing that covered 14 CpG sites. Our results showed that none of the examined BKV DNA from the various different stages of replication was methylated. This is the first report to analyze the methylation of BKV genomic DNA during viral replication. The results seem to indicate that methylation is not involved in regulation of BKV replication. PMID- 21626300 TI - Association between Epstein-Barr virus infection and chemoresistance to docetaxel in gastric carcinoma. AB - Epstein-Barr virus (EBV) is associated with human cancers such as nasopharyngeal carcinoma, Burkitt's lymphoma, Hodgkin's disease, and gastric carcinoma (GC). EBV is associated with about 10% of all GC cases globally. EBV-associated GC has distinct features from EBV-negative GC. However, it is still unclear if EBV infection has any effect on GC chemoresistance. Cell proliferation assay, cell cycle analysis, and active caspase Western blot revealed that the EBV-positive GC cell line (AGS-EBV) showed chemoresistance to docetaxel compared to the EBV negative GC cell line (AGS). Docetaxel treatment increased expression of Bax similarly in AGS and AGS-EBV cell lines. However, Bcl-2 induction was markedly higher in AGS-EBV cells, after docetaxel treatment. Although docetaxel increased the expression of p53 to a similar extent in both cell lines, induction of p21 in AGS-EBV cells was lower than in AGS cells. Furthermore, expression of survivin was higher in AGS-EBV cells than in AGS cells following docetaxel treatment as well as at basal state. EBVlytic gene expression was induced by docetaxel treatment in AGS-EBV cells. The results suggest that EBV infection and lytic induction confers chemoresistance to GC, possibly by regulating cellular and EBV latent and lytic gene expression. PMID- 21626301 TI - A new multidimensional model of successful aging: perceptions of Japanese American older adults. AB - This study examined the concept of successful aging using an ethnographic grounded-theory approach. Seventy-seven Japanese American older adults participated in focus groups. Participants perceived successful aging as optimal functioning in the following areas: Physical health, psychological health, cognitive functioning, socialization, spirituality, and financial security. The content of each dimension represents both culture-specific and culturally universal elements. This new multidimensional model of successful aging was compared to Rowe and Kahn's (The Gerontologist 37:433-440, 1997) and Phelan et al.'s frameworks (Journal of the American Geriatric Society 52:211-216, 2004) of successful aging. The model of successful aging generated from this study appears to be more comprehensive than existing models and incorporates sociocultural experiences. PMID- 21626302 TI - Co-variation among major classes of LRR-encoding genes in two pairs of plant species. AB - NBS-LRR (nucleotide-binding site-leucine-rich repeat), LRR-RLK (LRR-receptor-like kinase), and LRR-only are the three major LRR-encoding genes. Owing to the crucial role played by them in plant resistance, development, and growth, extensive studies have been performed on the NBS-LRR and LRR-RLK genes. However, few studies have focused on these genes collectively; they may co-vary as all of them contain LRR motifs. To investigate their common evolutionary patterns, all major classes of LRR-encoding genes were identified in 12 plant species, and particularly compared in two pairs of close relatives, Arabidopsis thaliana-A. lyrata (At-Al) and Zea mays-Sorghum bicolor. Our results showed that these genes co-vary significantly in terms of their numbers between species and that the genes with certain evolutionary parameters are most likely to have similar functions. The development-related genes have clear orthologous relationships between closely related species, as well as lower nucleotide divergence, and Ka/Ks ratio. In contrast, resistance-related genes have exactly opposite characteristics and favor 11-15 LRRs per gene. This association could be very useful in predicting the function of LRR-encoding genes. The presence of co variation suggests that LRRs, combined with other domains, can work better in some common functions. In order to cooperate efficiently, there should be balanced gene numbers among the different gene classes. PMID- 21626303 TI - Degradation of interleukin 8 by the serine protease MucD of Pseudomonas aeruginosa. AB - We investigated the influence of the type III effector, ExoS, on the host epithelial cell response to Pseudomonas aeruginosa infection, and we found that disruption of the exoS gene caused a significant increase in the amount of interleukin-8 (IL-8) in the culture medium of Caco-2 cells. We show that IL-8 was degraded in the culture medium following infection of the cells with the wild type (PAO1), but not the exoS knock-out (the DeltaexoS) strain. Purified ExoS protein itself did not degrade IL-8. We next show that IL-8 degradation by PAO1 was inhibited by the addition of serine protease inhibitors. These results strongly suggest that a bacterial serine protease that degrades IL-8 is expressed and secreted into the culture medium of Caco-2 cells infected with PAO1, and that the expression of this protein is repressed in cells infected with the DeltaexoS strain. The PAO1 genome encodes 28 different protease genes, including two serine proteases: PA3535 and mucD. PA3535 and mucD gene knock-outs were constructed (DeltamucD and DeltaPA3535), and DeltamucD but not DeltaPA3535 showed reduced IL 8 degradation. To understand the significance of IL-8 degradation, we next evaluated neutrophil infiltration in lungs excised from mice intranasally infected with the P. aeruginosa strains. Increased neutrophil infiltration was observed in PAO1-infected mice, but not in DeltaexoS- or DeltamucD-infected mice. Taken together, our results suggest that P. aeruginosa escapes from phagocytic killing due to IL-8 degradation following the secretion of the MucD serine protease, whose expression appears to be influenced by ExoS. PMID- 21626304 TI - Assessment of narrow angles by gonioscopy, Van Herick method and anterior segment optical coherence tomography. AB - PURPOSE: To evaluate anterior chamber (AC) angles using gonioscopy, Van Herick technique and anterior segment optical coherence tomography (AS-OCT). METHODS: One hundred forty-eight consecutive subjects were enrolled. The agreement between any two of three diagnostic methods, gonioscopy, AS-OCT and Van Herick, was calculated in narrow-angle patients. The area under receiver-operating characteristic curves (AUC) for discriminating between narrow and open angles determined by gonioscopy was calculated in all participants for AS-OCT parameter angle opening distance (AOD), angle recess area, trabecular iris surface area and anterior chamber depth (ACD). As a subgroup analysis, capability of AS-OCT parameters for detecting angle closure defined by AS-OCT was assessed in narrow angle patients. RESULTS: The agreement between the Van Herick method and gonioscopy in detecting angle closure was excellent in narrow angles (kappa = 0.80, temporal; kappa = 0.82, nasal). However, agreement between gonioscopy and AS-OCT and between the Van Herick method and AS-OCT was poor (kappa = 0.11-0.16). Discrimination capability of AS-OCT parameters between open and narrow angles determined by gonioscopy was excellent for all AS-OCT parameters (AUC, temporal: AOD500 = 0.96, nasal: AOD500 = 0.99). The AUCs for detecting angle closure defined by AS-OCT image in narrow angle subjects was good for all AS-OCT parameters (AUC, 0.80-0.94) except for ACD (temporal: ACD = 0.70, nasal: ACD = 0.63). CONCLUSION: Assessment of narrow angles by gonioscopy and the Van Herick technique showed good agreement, but both measurements revealed poor agreement with AS-OCT. The angle closure detection capability of AS-OCT parameters was excellent; however, it was slightly lower in ACD. PMID- 21626305 TI - Striking neuronal thymosin beta 4 expression in the deep layers of the mouse superior colliculus after monocular deprivation. AB - Thymosin beta 4 (Tbeta4), a small acidic peptide isolated originally from calf thymus, is recognized as a regulator of actin polymerization. It is widely distributed in the brain, in neurons as well as in microglia, and it has been implicated in lesion-induced neuroplasticity. In the present study, we quantitatively analyzed the immunocytochemical expression pattern of Tbeta4 in the superior colliculus (SC) of the adult mouse following monocular enucleation. Immunocytochemistry revealed a remarkable neuronal profile in the SC of 2, 5 and 7-week enucleated animals. We investigated the morphology, number and layer specific localization of these immunoreactive neurons. Neuronal Tbeta4 immunoreactive cell bodies were only sparsely found in the deep layers of the SC of control mice and showed only few and short immunoreactive branches, whereas their number increased significantly with post-enucleation survival time, as well as the total extent of the neuritic tree and its degree of branching. Occasionally, some Tbeta4-immunoreactive neurons were also detected in the periaqueductal gray. As a regulator of actin metabolism, Tbeta4 may influence the remodeling of these collicular neuronal processes, possibly in the context of a re-alignment of the sensory and motor maps in the deep SC as a consequence of the visual deprivation paradigm. PMID- 21626306 TI - Optic flow estimation on trajectories generated by bio-inspired closed-loop flight. AB - We generated panoramic imagery by simulating a fly-like robot carrying an imaging sensor, moving in free flight through a virtual arena bounded by walls, and containing obstructions. Flight was conducted under closed-loop control by a bio inspired algorithm for visual guidance with feedback signals corresponding to the true optic flow that would be induced on an imager (computed by known kinematics and position of the robot relative to the environment). The robot had dynamics representative of a housefly-sized organism, although simplified to two-degree-of freedom flight to generate uniaxial (azimuthal) optic flow on the retina in the plane of travel. Surfaces in the environment contained images of natural and man made scenes that were captured by the moving sensor. Two bio-inspired motion detection algorithms and two computational optic flow estimation algorithms were applied to sequences of image data, and their performance as optic flow estimators was evaluated by estimating the mutual information between outputs and true optic flow in an equatorial section of the visual field. Mutual information for individual estimators at particular locations within the visual field was surprisingly low (less than 1 bit in all cases) and considerably poorer for the bio-inspired algorithms that the man-made computational algorithms. However, mutual information between weighted sums of these signals and comparable sums of the true optic flow showed significant increases for the bio-inspired algorithms, whereas such improvement did not occur for the computational algorithms. Such summation is representative of the spatial integration performed by wide-field motion-sensitive neurons in the third optic ganglia of flies. PMID- 21626307 TI - GABA and GAD expression in the X-organ sinus gland system of the Procambarus clarkii crayfish: inhibition mediated by GABA between X-organ neurons. AB - In crustaceans, the X-organ-sinus gland (XO-SG) neurosecretory system is formed of distinct populations of neurons that produce two families of neuropeptides: crustacean hyperglycemic hormone and adipokinetic hormone/red pigment concentrating hormone. On the basis of electrophysiological evidence, it has been proposed that gamma-aminobutyric acid (GABA) regulates both electrical and secretory activity of the XO-SG system. In this work we observed that depolarizing current pulses to neurons located in the external rim of the X-organ induced repetitive firing that suppressed the spontaneous firing of previously active X-organ neurons. Picrotoxin reversibly blocked this inhibitory effect suggesting that the GABA released from the stimulated neuron inhibited neighboring cells. Immunoperoxidase in X-organ serial sections showed co localization of GABA and glutamic acid decarboxylase (GAD) including the aforementioned neurons. Immunofluorescence in whole mount preparations showed that two subpopulations of crustacean hyperglycemic hormone-containing neurons colocalized with GABA. The expression of GAD mRNA was determined in crayfish tissue and X-organ single cells by RT-PCR. Bioinformatics analysis shows, within the amplified region, 90.4% consensus and 41.9% identity at the amino acid level compared with Drosophila melanogaster and Caenorhabditis elegans. We suggest that crustacean hyperglycemic hormone-GABA-containing neurons can regulate the excitability of other X-organ neurons that produce different neurohormones. PMID- 21626308 TI - Impact of hypertension on stroke. AB - Hypertension is the single most important risk factor for all types of stroke: ischemic stroke, intracerebral hemorrhage, and aneurysmal subarachnoid hemorrhage. Epidemiologic studies over the past 30 years have demonstrated a dramatic reduction in the incidence and mortality of all stroke types with good control of hypertension, and it appears that all effective antihypertensive agents have similar efficacy in their ability to reduce stroke risk. In addition, it appears that acute treatment of hypertension in the setting of intracerebral hemorrhage and subarachnoid hemorrhage is beneficial, but it is still uncertain in the setting of ischemic stroke what level of blood pressure will result in the best possible outcome. PMID- 21626309 TI - Determination of composition and structure of spongy bone tissue in human head of femur by Raman spectral mapping. AB - Biomechanical properties of bone depend on the composition and organization of collagen fibers. In this study, Raman microspectroscopy was employed to determine the content of mineral and organic constituents and orientation of collagen fibers in spongy bone in the human head of femur at the microstructural level. Changes in composition and structure of trabecula were illustrated using Raman spectral mapping. The polarized Raman spectra permit separate analysis of local variations in orientation and composition. The ratios of nu2PO43-/Amide III, nu4PO43-/Amide III and nu1CO32-/nu2PO43- are used to describe relative amounts of spongy bone components. The nu1PO43-/Amide I ratio is quite susceptible to orientation effect and brings information on collagen fibers orientation. The results presented illustrate the versatility of the Raman method in the study of bone tissue. The study permits better understanding of bone physiology and evaluation of the biomechanical properties of bone. PMID- 21626310 TI - Perivascular epithelioid cell tumors (PEComas) of the head and neck: report of three cases and review of the literature. AB - PEComas are a family of neoplastic lesions that share overlapping morphology, immunohistochemistry, and ultrastructure that include angiomyolipoma, lymphangioleiomyomatosis, clear cell "sugar" tumor of the lung as well as similar tumors occurring in a variety of visceral, cutaneous and soft tissue sites throughout the body. The defining histopathological features are epithelioid cells with a perivascular distribution containing clear to pale eosinophilic granular cytoplasm and a round-to-oval centrally located nucleus with an inconspicuous nucleolus. Immunohistochemically, coexpression of melanocytic (HMB 45 and/or Melan-A) and myoid markers are characteristic. In the present study, we describe three PEComas occurring in the head and neck (nasal cavity and larynx) and discuss the behavior of these distinctive tumors and review the literature of head and neck PEComas. The importance of recognizing this entity will ensure its consideration in the differential diagnosis of tumors of the head and neck with a similar morphology. The histogenesis of PEComas still remains elusive and additional cases with a prolonged follow up remain important to accurately determine the behavior of these distinctive tumors. Complete surgical excision still remains the treatment of choice for histologically benign PEComas. PMID- 21626311 TI - Association between effort-reward imbalance and glycosylated hemoglobin (HbA1c) among Chinese workers: results from SHISO study. AB - OBJECTIVE: The studies focusing on effort-reward imbalance and diabetes mellitus (DM)/glycosylated hemoglobin (HbA1c) are rare. We sought to examine the association between job stress evaluated by effort-reward imbalance (ERI) model and HbA1c in a Chinese population. METHODS: We analyzed 680 subjects (465 men and 215 women) without DM or impaired glucose tolerance from the stress and health in Shenzhen workers (SHISO) study. Job stress was evaluated by effort-reward imbalance (ERI) model. HbA1c was measured by an automatic analyzer by means of high-performance liquid chromatography. The association between job stress and HbA1c was explored by variance analysis, partial correlations and multiple linear regression analysis. RESULTS: For women, effort, and ERI were positively associated with HbA1c (r = 0.22, p = 0,003; r = 0.21, p = 0.006, respectively), in contrast, reward was negatively associated with HbA1c (r = -0.17, p = 0.021), after controlling age, BMI and physical exercise in the partial correlation analysis; the similar results were confirmed in the multiple linear regression. No significant correlations between job stress and HbA1c were found for men. CONCLUSION: Effort and ERI are positively associated with HbA1c, and reward is inversely related to HbA1c among Chinese women. The association is not accounted for by age, BMI, and physical exercise. More efforts should be made to improve the job stress status of Chinese working women for the purpose of DM prevention. PMID- 21626312 TI - Impaired work functioning due to common mental disorders in nurses and allied health professionals: the Nurses Work Functioning Questionnaire. AB - PURPOSE: Common mental disorders (CMD) negatively affect work functioning. In the health service sector not only the prevalence of CMDs is high, but work functioning problems are associated with a risk of serious consequences for patients and healthcare providers. If work functioning problems due to CMDs are detected early, timely help can be provided. Therefore, the aim of this study is to develop a detection questionnaire for impaired work functioning due to CMDs in nurses and allied health professionals working in hospitals. METHODS: First, an item pool was developed by a systematic literature study and five focus group interviews with employees and experts. To evaluate the content validity, additional interviews were held. Second, a cross-sectional assessment of the item pool in 314 nurses and allied health professionals was used for item selection and for identification and corroboration of subscales by explorative and confirmatory factor analysis. RESULTS: The study results in the Nurses Work Functioning Questionnaire (NWFQ), a 50-item self-report questionnaire consisting of seven subscales: cognitive aspects of task execution, impaired decision making, causing incidents at work, avoidance behavior, conflicts and irritations with colleagues, impaired contact with patients and their family, and lack of energy and motivation. The questionnaire has a proven high content validity. All subscales have good or acceptable internal consistency. CONCLUSION: The Nurses Work Functioning Questionnaire gives insight into precise and concrete aspects of impaired work functioning of nurses and allied health professionals. The scores can be used as a starting point for purposeful interventions. PMID- 21626313 TI - The interrelating dynamics of hypoxic tumor microenvironments and cancer cell phenotypes in cancer metastasis. AB - The interrelating dynamics of the primary tumor cells and their surrounding microenvironment might determine phenotypic characteristics of disseminated tumor cells and contribute to cancer metastasis. Cytoprotective mechanisms (e.g., energy metabolism control, DNA damage response, global translation control and unfolded protein response) exert selective pressure in the tumor microenvironment. In particular, adaptation to hypoxia is vital for survival of malignant cells in the tumor and at distant sites such as the bone marrow. In addition to the stress response, the ability of tumor cells to undergo certain cellular re-differentiation programmes like the epithelial-mesenchymal transition (EMT), which is linked to cancer stemness, appears to be important for successful cancer cell spread. Here we will discuss the selection pressures that eventually lead to the formation of overt metastases. We will focus the properties of the microenvironment including (i) metabolic and cytoprotective programs that ensure survival of disseminated tumor cells, (ii) blood vessel structure, and (iii) the hypoxia-normoxia switch as well as intrinsic factors affecting the evolvement of novel tumor cell populations. PMID- 21626317 TI - Esophageal reconstruction with colon tissue. AB - The present best practice for performing esophageal reconstruction using colon tissue was investigated in this review. The left colon has advantages in that it has less variation in blood supply and a smaller diameter than the right colon; however, the rate of graft necrosis is higher for the left colon. Additional microvascular anastomosis, which is unnecessary in most cases, may be able to resolve these issues. The colon graft should be reconstructed in an isoperistaltic fashion whenever possible in order to prevent regurgitation and improve food transit. The posterior mediastinum has the advantage of being the shortest route, but it also has the major disadvantage that graft necrosis can be severe or fatal if it occurs. In palliative or advanced cases, a retrosternal or subcutaneous route is preferred, because the posterior mediastinum is a tumor bed. However, in these cases partial excision of the manubrium and the left clavicula should be considered to release compression of the graft at the thoracic inlet. Consequently, the selection of the colon graft should be flexible and be based on the inspection of blood supply and the length needed, and thereafter microvessel anastomosis should be added in cases where graft ischemia might occur. PMID- 21626318 TI - Direct hemoperfusion with polymyxin B immobilized fiber for abdominal sepsis in Europe. AB - Since direct hemoperfusion with polymyxin B immobilized fiber (PMX-DHP) received its product certification for use in Europe in 1998, several prospective randomized controlled trials (RCTs) have been conducted in European countries. The first RCT, performed in six European academic medical centers in 2005, concluded that PMX-DHP is associated with improved hemodynamic status and cardiac function. Subsequently, a meta-analysis of PMX-DHP was presented in Italy in 2007. This systematic review found positive effects of PMX-DHP on mean arterial pressure and dopamine/ dobutamine use, PaO2/FiO2 ratio, endotoxin removal, and mortality. However, like most trials on extracorporeal therapies, none of the studies was double-blinded. The EUPHAS study, a multicenter RCT performed in ten Italian intensive care units in 2009, found that PMX-DHP improved 28-day survival, blood pressure, vasopressor requirement, and degree of organ failure. However, investigators in Belgium and Canada pointed out that there was no statistical difference in 28-day survival. Two more RCTs, the ABDO-MIX and EUPHRATES studies, the primary end points of which are 28-day mortality, were started in Europe and the United States at the end of 2010. We are hoping that these RCTs will resolve this issue. PMID- 21626319 TI - Which is the best technique for pancreaticoenteric reconstruction after pancreaticoduodenectomy? A critical analysis. AB - The definitive surgical management of periampullary tumors is a challenging endeavor. This article reviews the available data on the efficacy of various methods of pancreaticoenteric reconstruction designed for the prevention of pancreatic fistula (PF). A literature search of the Medline database was used to identify randomized controlled trials (RCTs) that compared pancreaticojejunostomy (PJ) with pancreaticogastrostomy (PG) after pancreaticoduodenectomy (PD). A total of two metaanalyses and four prospective RCTs were identified. Individual RCTs comparing PJ and PG allow the surgeons participating in the trial to choose technical modifications of one particular technique. As a result, there is no universal agreement as to whether one particular variation is safer and less prone to PF than the others. In addition, the majority of RCTs failed to stratify patient risk of PF. Further studies are therefore necessary to define the optimal technique of pancreatic reconstruction after PD conducted in high-volume centers by high-volume surgeons. PMID- 21626320 TI - Thyroidectomy with LigaSure. AB - PURPOSE: A new method for ensuring hemostasis during thyroid surgery has recently been introduced. This technique, electrothermal (LigaSure) and ultrasound blood vessel sealing, is still experimental. The aim of the present study was to evaluate the applications and efficacy of LigaSure by analyzing the duration of the surgery and the rate of complications of the surgical procedure. METHODS: A total of 23 patients who underwent the LigaSure operation (n(LS) group) were analyzed in a prospective nonrandomized/partly randomized clinical study, and were compared with patients who underwent treatment using the conventional (ligature) surgical technique. RESULTS: At our institutions, 187 patients were treated surgically for thyroid disease in 2006, and 23 of these patients were treated using LigaSure. The complication rate in the patients treated with LigaSure (n(LS): 4.3%) was lower than those who received conventional ligature surgery. However, given the small number of patients, this difference was not statistically significant (retrospective group n(1): 10.7%; nonrandomized group n(2): 9.8%; and randomized group n(3): 9.1%). The length of surgery in the LigaSure group (n(LS) = 65 +/- 3 min) was significantly shorter (P < 0.001) compared with both nonrandomized (n(2) = 71 +/- 6 min) and randomized (n(3) = 70 +/- 4 min) patients who received a conventional ligature. CONCLUSIONS: The application of LigaSure using meticulous surgical technique provides a new option for a safe thyroidectomy. Moreover, the duration of the LigaSure procedure is significantly shorter, and there are fewer complications as compared with the classic surgical thyroidectomy technique. PMID- 21626321 TI - Feasibility and safety of postoperative management without chest tube placement after thoracoscopic wedge resection of the lung. AB - PURPOSE: The aim of the present study was to assess the feasibility and safety of several improved criteria to avoid chest tube placement after thoracoscopic wedge resection of the lung. METHODS: From 2000 to 2009, 333 patients who underwent thoracoscopic wedge resections of the lung were reviewed. The patients were classified into two groups: (1) the no chest tube group (NCT), consisting of 132 patients in whom chest tubes were not placed because no air leakage or bleeding during intraoperative alternative sealing test was confirmed, and (2) the chest tube placement (CTP) group, consisting of 201 patients in whom chest tubes were placed because the criteria for the nonplacement of a chest tube were not met. The clinical data and postoperative morbidity were assessed between the two groups. RESULTS: The number of specimens (1.3 vs 1.5) and the endostapler cartridges used (2.5 vs 3.3), and the duration of the postoperative hospital stay (4.6 vs 6.7 days) in the NCT group were significantly lower than in the CTP group. One patient from the NCT group required chest tube insertion due to the development of late pneumothorax. However, no significant differences were found between the two groups. CONCLUSIONS: Our improved criteria are therefore considered to positively contribute to a safe and definite clinical decision regarding postoperative patient management. PMID- 21626322 TI - Selection of pulmonary resection procedures to reduce postoperative complications in 200 patients. AB - PURPOSE: We previously demonstrated in a pilot study that postoperative cardiopulmonary complications could be reduced by selecting pulmonary resection procedures based on the results of a combination of specific preoperative cardiopulmonary function tests. The present study reports a re-examination of the criteria for preoperative screening and prospectively assessed whether the selected surgical procedures were appropriate in 200 patients who underwent the planned extent of pulmonary resection. METHODS: In 200 patients requiring lung tumor resection, five preoperative parameters (forced expiratory volume in 1 s on the intact side, maximal oxygen uptake, ejection fraction, occluded pulmonary artery pressure, and occluded total pulmonary vascular resistance index) were used to assign each patient to one of five risk categories in order to select the optimal resection procedure. Thereafter, the postoperative course was investigated to determine the value of this selection method. RESULTS: Thoracotomy was performed in 195 of the 200 patients (97.5%). Two patients (1%) died; one patient succumbed to acute exacerbation of interstitial pneumonia and the other patient died from pulmonary embolism. Six patients (3.1%) developed major complications after surgery and 12 patients (6.2%) had mild complications, while 175 (89.7%) showed a good postoperative course. CONCLUSION: The use of five preoperative parameters to select the pulmonary resection procedure minimized postoperative death and major complications. PMID- 21626323 TI - Assessing a clinical pathway to improve the quality of care in pulmonary resections. AB - PURPOSE: To evaluate the efficacy of the current clinical pathway for pulmonary resections. METHODS: This study examined variances from expected clinical pathway outcomes for pulmonary resections performed between 2005 and 2009. Data on a total of 383 patients were retrospectively analyzed. RESULTS: The median length of hospital stay (LOS) using the clinical pathway was 12 days (range: 1-188 days); the mean LOS was 15.5 days. The cost per day with use of the clinical pathway was 102 726 yen. Poor control of pain from intercostal neuralgia was the most frequently observed variance from expected outcomes. It affected 119 of 168 electronic clinical pathway patients (70.8%). The clinical pathway was terminated in 3.9% of patients (15/383) due to serious or life-threatening complications. CONCLUSIONS: This study showed the single institutional experience of the clinical pathway for pulmonary resections. These findings indicate a need to revise certain aspects of the pathway, based on data from our analysis of variances. PMID- 21626324 TI - Efficacy of prophylactic treatment with selective serotonin reuptake inhibitors for depression after open-heart surgery. AB - PURPOSE: The aim of the present study was to evaluate the efficacy of prophylactic treatment with selective serotonin reuptake inhibitors (SSRIs) in female patients at high risk of suffering depression after cardiac surgery. METHODS: Female patients (n = 58; group I) who were over 70 years of age or who had undergone emergency surgery were administered prophylactic treatment with paroxetine immediately after surgery. The hospital mortality and morbidity data of these patients were compared with those of 59 patients (group II) without prophylactic medication. RESULTS: The Center for Epidemiological Studies Depression Scale (CES-D) score at 10 days after surgery was significantly lower in group I (15.2 +/- 7.8) than in group II (21.8 +/- 7.5, P = 0.0018). The incidence of depression (I: 12.1% vs II: 64.4%, P < 0.0001) and pneumonia (I: 0% vs II: 10.2%, P = 0.0127) were significantly lower in group I than in group II. In addition, the duration of postoperative hospital stay was significantly shorter for group I than for group II (I: 15.9 +/- 56.5 vs II: 23.4 +/- 20.5 days, P = 0.0102). The hospital mortality rates were similar. CONCLUSION: The quality of life of patients with depression after open-heart surgery is poor. The early administration of prophylactic medication is therefore necessary for those patients at risk for developing depression. PMID- 21626325 TI - Foreign bodies in the rectum: an analysis of 30 patients. AB - PURPOSE: A foreign body in the rectum is a rare clinical entity. These foreign bodies can be classified under two major groups: the ones that are inserted from the anus and the ones that are swallowed and thereafter become stuck in the rectum. METHODS: Data of patients admitted with a foreign body in the rectum between 1988 and 2008 were retrospectively analyzed. A total of 30 patients were reviewed. RESULTS: The most common reason for a foreign body in the rectum was insertion for erotic purposes, which was seen in 19 of the 30 patients. All these patients were male. The remaining 11 patients reported nonerotic causes. The most efficient and implemented therapeutic method was simple extraction during the first examination. CONCLUSIONS: The most common reason for a foreign body in the rectum is insertion for erotic purposes. The first target of therapy should be extraction of the foreign body using the simplest method possible. Meanwhile, protecting the integrity of the intestine is of the utmost importance. The care for maintaining the integrity of the rectum should include an evaluation of the patient's psychological status. PMID- 21626326 TI - Trends in the treatment outcomes for advanced colorectal cancer: an analysis at a single community hospital in Japan. AB - PURPOSE: To evaluate the trends in the treatment outcomes for patients with colorectal cancer in Japan. METHODS: We performed a retrospective analysis of patients undergoing surgery for primary colorectal cancer during a 16-year period (Group A [1992-2000, n = 258]; Group B [2001-2008, n = 258]) at Fukuoka City Hospital. Because no significant differences were found in the survival rates in stage 0, I, II, and III patients between the two groups, we concentrated on examining stage IV patients. RESULTS: The 3-year survival rate for stage IV patients in Group B (n = 26) was significantly higher than that in Group A (n = 31) (34.9% vs 3.9%, P < 0.05). The rate of curative resection for advanced liver metastases in Group B patients was also significantly higher than that of Group A patients (50.0% vs 13.3%, P < 0.05). As a result, the 2-year survival rate for the disease-free patients in Group B was significantly higher than that for the non-disease-free patients in Group B (46.0% vs 21.0%, P < 0.05). Group B had a greater proportion of patients receiving l-leucovorin/5-fluorouracil than Group A (8 patients vs none, P < 0.05). CONCLUSIONS: Recent advances in surgical innovations and the utilization of new chemotherapeutic agents may have led to significant improvements in the treatment outcomes for advanced colorectal cancer in Japan. PMID- 21626327 TI - Aluminum potassium sulfate and tannic acid (ALTA) injection as the mainstay of treatment for internal hemorrhoids. AB - PURPOSE: Aluminum potassium sulfate and tannic acid (ALTA) induce noninvasive sclerosis and the involution of hemorrhoids by initiating an inflammatory reaction. We assessed the mid-term outcome after ALTA sclerotherapy for symptomatic hemorrhoids. METHODS: Between May 2006 and July 2009, 1210 patients with grade III or IV hemorrhoids underwent surgery at Kunimoto Hospital. Our treatment strategy for internal hemorrhoids is first establishing whether ALTA therapy is possible for the type of hemorrhoid, and then performing either ALTA therapy or alternatively, ligation and excision (LE) for those types unsuitable for ALTA therapy. RESULTS: A total of 448 patients were treated with ALTA therapy alone (Group A), 706 patients were treated with a combination of ALTA and LE therapy (Group B), and 56 patients were treated with LE alone (Group C). The overall recurrence rates were 3.6% (16/448) and 0.3% (2/706) in Groups A and B, respectively. There was no recurrence in Group C. Rectal ulcers developed at the injection site in four (0.9%) patients from Group A, but they healed within a few months with conservative therapy. CONCLUSION: ALTA sclerotherapy is a simple and safe treatment for symptomatic hemorrhoids, with few complications. PMID- 21626328 TI - Combined lysosomal protein transmembrane 4 beta-35 and argininosuccinate synthetase expression predicts clinical outcome in hepatocellular carcinoma patients. AB - PURPOSE: The newly-identified lysosomal protein transmembrane 4 beta-35 (LAPTM4B 35) plays important roles in tumor progression and metastasis, while argininosuccinate synthetase (ASS) provides arginine as an indispensable nutrient for hepatocellular carcinoma (HCC). The present study investigated the clinical significance of the coexpression of LAPTM4B-35 and ASS in HCC patients on determining the prognosis. METHODS: Immunohistochemistry was used to evaluate the expression of LAPTM4B-35 and ASS in HCC tissues and paired noncancerous liver samples from 71 patients. The correlation of combined LAPTM4B-35 and ASS expression with selected clinicopathologic parameters was assessed with the chi squared test. Patient survival and differences in survival were determined by the Kaplan-Meier method and the log-rank test. A Cox regression analysis was adopted for a multivariate analysis of the prognostic factors. RESULTS: Combined LAPTM4B 35 and ASS expression was significantly associated with TNM stage and portal vein invasion. In addition, patients with HCCs expressing both LAPTM4B-35 and ASS exhibited both markedly poorer overall survival (OS) and disease-free survival (DFS) (both P < 0.001). According to the multivariate analyses, combined LAPTM4B 35 and ASS expression was found to be an independent prognostic factor for OS and DFS (P = 0.039 and P = 0.035, respectively). CONCLUSION: The overexpression of LAPTM4B-35 in combination with positive ASS expression is a negative prognostic marker for HCC. PMID- 21626329 TI - The influence of intracellular epidermal growth factor receptor (EGFR) signal activation on the outcome of EGFR tyrosine kinase inhibitor treatment for pulmonary adenocarcinoma. AB - PURPOSE: The epidermal growth factor receptor tyrosine kinase inhibitors (EGFR TKIs), such as gefitinib, exhibit up to a 70% response rate against non-small cell lung cancer (NSCLC) harboring somatic activating mutations of the EGFR gene (EGFR). The mechanism of intrinsic resistance of EGFR mutation-positive NSCLC against EGFR-TKIs is not known. The current study assesses the relationship between the molecular expression of EGFR signals and the response to gefitinib treatment in patients with pulmonary adenocarcinoma to elucidate the mechanism of intrinsic resistance to gefitinib. METHODS: The present study included 30 patients with pulmonary adenocarcinoma who were treated with gefitinib for a postoperative recurrence. The correlation between the response to gefitinib treatment and various clinical and molecular features was evaluated. RESULTS: EGFR mutations were detected in 20 (66.7%) of the 30 patients. The response to gefitinib treatment was a complete response in 1 case, partial response in 12 cases, stable disease in 4 cases, and progressive disease in 13 cases. Both univariate and multivariate analyses showed the presence of an EGFR mutation, and the expression of phospho-EGFR (p-EGFR) significantly correlated with a better response to gefitinib treatment. Ten of the 16 p-EGFR positive patients were disease controlled, but all 4 p-EGFR negative patients were intrinsically resistant to EGFR-TKIs (P = 0.025). Other factors including sex, smoking status, serum carcinoembryonic antigen and cytokeratin-19 fragment levels, EGFR, Met proto-oncogene, phospho-Met, and hepatocyte growth factor expression were not associated with the response to gefitinib treatment. CONCLUSION: These results suggest that, even if EGFR mutations were observed, a p-EGFR negative state might be a cause of intrinsic resistance to EGFR-TKIs. PMID- 21626330 TI - Mechanical intestinal cleansing and antibiotic prophylaxis for preventing bacterial translocation during the Pringle maneuver in rabbits. AB - PURPOSE: We investigated the effectiveness of mechanical intestinal cleansing and antibiotic prophylaxis in preventing bacterial translocation (BT) during the Pringle maneuver in rabbits. METHODS: Forty-eight rabbits were allocated to one of the following four groups: a control group (group 1); an antibiotic group, given 100 mg/kg intravenous ceftizoxime (group 2); a mechanical intestinal cleansing group, given a Fleet enema (group 3); and a mechanical intestinal cleansing plus antibiotic group (group 4). After performing laparotomy, we dissected the portal region and turned the portal triad, using tape. Pringle maneuver was applied for 30 min in all groups. Blood samples were collected from the portal vein for blood culture before the Pringle maneuver. All groups underwent relaparotomy 30 min after the Pringle maneuver, to obtain portal blood, mesenteric lymph nodes (MLNs), and splenic tissue for culture. RESULTS: All cultures from the portal vein specimens taken before the Pringle maneuver were negative. The rate of bacterial isolation in the portal vein (P < 0.001), MLNs (P < 0.01), and splenic (P < 0.001) cultures was significantly lower in group 4 than in the other groups. It was also lower in group 3 than in groups 1 and 2 (P < 0.05 for all). CONCLUSIONS: The combination of mechanical intestinal cleansing and preoperative broad-spectrum antibiotics was most effective for preventing BT during the Pringle maneuver. PMID- 21626331 TI - Primary neuroendocrine carcinoma of the breast: report of a case. AB - Primary neuroendocrine carcinoma (NEC) of the breast appears to be a rare neoplasm. Due to the limited number of the cases, a definitive therapeutic option for the disease has not yet been established. We herein report the case of a 57 year-old female patient with primary NEC of the breast who underwent a surgical resection and for whom the suitable adjuvant therapy is now being considered. PMID- 21626332 TI - Esthetic result of rhomboid flap repair after breast-conserving surgery for lower quadrant breast cancer lesion with skin invasion: report of two cases. AB - Breast-conserving surgery (BCS) has been increasingly performed as a standard operative strategy for patients with breast cancer. The primary purpose of BCS is to acquire both local control and good cosmetic results. An insignificant difference in cancer treatment results has been shown between BCS and total mastectomy. However, achieving sufficiently cosmetic results can be difficult, particularly in patients with tumors that are large or localized to the lower quadrant. To avoid breast deformities and asymmetries after BCS, immediate reconstruction using autologous tissue has been accepted as the standard option. Rhomboid skin and adipose flap repair is a simple, less invasive procedure than the myocutaneous flap, which has primarily been performed in patients with upper quadrant lesions. We herein report the cases of two patients with lower quadrant breast cancer with skin invasion, who underwent BCS with immediate breast repair using a rhomboid flap. This procedure is therefore worth considering as one of the first options for immediate repair after BCS. PMID- 21626333 TI - Sentinel lymph node biopsy in patients with male breast carcinoma: report of two cases. AB - The incidence of male breast cancer is low, and treatment strategies similar to those used for female breast cancer patients are frequently used for male patients. However, the safety and utility of sentinel lymph node biopsies (SLNBs) for male breast cancer have not been proven. Among the five cases of male breast cancer who received surgery at our hospital, mastectomy with SLNB was performed in two of the cases. The first patient was 77 years old and the second was 74 years old, and both presented as outpatients with chief complaints of a mammary mass. Clinical diagnoses were T1N0 in both cases, and mastectomies with SLNB were performed. The sentinel lymph node was identified using the dye method. Postoperatively, the patients were hormone receptor-positive, and they are now being followed while continuing to take oral tamoxifen. PMID- 21626334 TI - Pleomorphic carcinoma of the lung arising in a patient with Li-Fraumeni syndrome: report of a case. AB - We herein report the case of a patient with Li-Fraumeni syndrome (LFS) who developed lung pleomorphic carcinoma. A 28-year-old female patient with a family history of early-onset malignancies was diagnosed with lung carcinoma and treated by surgical resection. Histological examination revealed a heterogeneous tumor with epithelial and mesenchymal components. The final pathological diagnosis was pulmonary pleomorphic carcinoma. In this patient, a constitutional mutation at codon 213 in exon 6 of the p53 gene was identified in the peripheral lymphocytes and the resected tumor, and LFS was suspected. This mutation causes a nonsense mutation (Arg-to-Stop codon) that has been shown to attenuate p53 function. This is the first report of pulmonary pleomorphic carcinoma developing in an LFS patient, and may suggest a relationship between germline p53 mutation and carcinogenesis in pulmonary pleomorphic carcinoma. PMID- 21626335 TI - Arteriovenous hemangioma in the middle mediastinum: report of a case. AB - Hemangiomas in the mediastinum are uncommon, and the majority of these tumors are located in the anterior mediastinum. The present report describes an extremely rare case of a hemangioma in the middle mediastinum. A 40-year-old male patient presented with a mass in the left lower lung field on radiographic screening. Chest computed tomography and magnetic resonance imaging revealed a mass measuring 2.5 cm in diameter, which was close to the descending aorta in the left middle mediastinum. The tumor resection was performed by video-assisted thoracic surgery. The thoracoscopic findings revealed a blackish brown mass with a capsule, which had not invaded the descending aorta or cardiac sac. Histologically, the tumor was composed of typical thick-walled vessels and was therefore classified as an arteriovenous hemangioma. PMID- 21626336 TI - Pulmonary resection for lung cancer with impaired pulmonary function due to severe spinal deformity: report of a case. AB - A 58-year-old man underwent surgery for the treatment of lung adenocarcinoma at our hospital. He had a history of spinal caries at the age of 3 and frequent recurrent asthma-like symptoms thereafter. Chest plain radiography revealed an irregularly shaped mass in the left hilum and a thoracic cage severely deformed by kyphosis. A pulmonary function test demonstrated a severe restrictive ventilation pattern. He underwent a left upper lobectomy without mediastinal dissection. His asthma-like symptoms were exacerbated by difficulty in expectoration of sputum for several days after surgery, but the symptoms disappeared without any medical care. Dyspnea, hemoptysis, and intrathoracic hemorrhage were observed on the 8th postoperative day. Computed tomography revealed torsion of the residual left lower lobe, but the symptoms spontaneously disappeared in a week. The patient had complained of frequent coughing and mild dyspnea on exertion for years, but his quality of life was unchanged after the lobectomy. Bone metastasis was detected 3 years after the surgery, and the patient then underwent chemotherapy. However, he died of the disease 4 years 2 months after the surgery. PMID- 21626337 TI - Metastatic splenic alpha-fetoprotein-producing adenocarcinoma: report of a case. AB - Solitary carcinomatous metastases to the spleen are rare, and the presence of solitary splenic metastasis of hepatoid adenocarcinomas (HAC) has not yet been reported. We herein present the case of a 54-year-old male patient who developed a metastatic tumor to the spleen with enhanced levels of serum alpha-fetoprotein and carcinoembryonic antigen (CEA). The patient underwent a gastrectomy due to adenocarcinoma of the stomach 5 years previously. The tumor was diagnosed on the grounds of histopathology and immunohistochemical staining after splenectomy. The pathology slides showed both adenocarcinoma and hepatoid structures. alpha Fetoprotein antibody staining was positive in the tumor, suggesting hepatoid characteristics. After the operation, the patient recovered well and had no signs of recurrence at a follow-up examination after 9 months. Together with this case, several aspects of the disease are discussed after a review of the current literature. PMID- 21626338 TI - Small intestinal metastasis from intrahepatic cholangiocarcinoma: report of a case. AB - Metastasis of the small intestine that derives from a primary hepatic neoplasm is rare. We encountered a case of intrahepatic cholangiocarcinoma (ICC) with jejunal metastasis after resection of a primary lesion. A 61-year-old male patient was referred to us with a diagnosis of liver tumors. Partial hepatectomy was performed, and the pathological diagnosis was ICC. Seventeen months after surgery, the patient was found to have a mass in the jejunum and lymph node swelling by positron emission tomography-computed tomography. The jejunal tumor was preoperatively diagnosed as a metastasis of ICC from a biopsy specimen obtained by double balloon endoscopy, and the tumor was resected. The patient received systemic chemotherapy but succumbed with ICC recurrence 46 months after the primary surgery. To the best of our knowledge, this case is the first report of jejunal recurrence of ICC. In addition, this report suggests the usefulness of double balloon endoscopy to make the correct diagnosis of the jejunal tumor. PMID- 21626339 TI - Synchronous solid pseudopapillary neoplasm and intraductal papillary mucinous neoplasm of the pancreas: report of a case. AB - Solid pseudopapillary neoplasm (SPN) of the pancreas, most commonly found in young female subjects, is a rare neoplasm with low potential for malignancy. We report an unusual case of a 66-year-old male patient who had a simultaneous malignant SPN and an intraductal papillary mucinous adenoma (IPMA) of the pancreas. The patient was admitted to our department for the evaluation of the main solid tumor with calcification and small multilocular cystic lesions apart from the main tumor in the pancreatic head. We performed pylorus-preserving pancreaticoduodenectomy to treat the calcified tumor and multilocular cystic lesions. The diagnosis of malignant SPN was confirmed on the basis of histological invasion to the adjacent structures. The separate cystic lesions were diagnosed as a branch-type IPMA. The synchronous occurrence of IPMA and SPN in the present case did not demonstrate that there were tumors maintained through the common abnormal Wnt signaling pathway by immunohistochemical study. To our knowledge, this is the first known case of synchronous SPN and IPMA of the pancreas. PMID- 21626340 TI - Pancreatic adenosquamous carcinoma presenting as splenic rupture: report of a case. AB - A 58-year-old female patient presented with the sudden onset of left upper quadrant pain. The physical examination revealed the presence of shock status. Abdominal computed tomography revealed splenomegaly with a huge mass inside the spleen, and massive fluid collection in the abdominal cavity. After splenic artery embolization, laparotomy was performed. The operative findings revealed intra-abdominal hemorrhage and rupture of the lower pole of the spleen. Furthermore, a palpable solid mass was observed at the splenic hilum, and distal pancreatectomy with splenectomy was performed. The macroscopic findings revealed a pancreatic tail tumor at the splenic hilum directly invading the splenic parenchyma. Microscopic examinations showed the tumor to consist of squamous cell carcinoma. Furthermore, old and new thrombi were observed inside small splenic arteries. These findings were considered to represent invasion of pancreatic adenosquamous carcinoma to the spleen, and rupture of the spleen was attributed to splenic ischemia resulting from cancer invasion and splenic vein obstruction. PMID- 21626341 TI - Single-incision multiport laparoscopic cholecystectomy for a patient with situs inversus totalis: report of a case. AB - Laparoscopic cholecystectomy has become the standard treatment for symptomatic cholelithiasis in patients with situs inversus totalis (SIT). Nowadays, single incision multiport laparoscopic surgery is safe and feasible for treating benign gallbladder disease. We report a case of successful single-incision multiport laparoscopic cholecystectomy for a patient with SIT, and describe its technical advantages. PMID- 21626342 TI - Pigmented hepatocellular adenoma: report of a case. AB - Hepatocellular adenoma is a rare, benign neoplasm that is related to the use of contraceptives or androgenic-anabolic steroids. A 51-year-old man with no history of steroid use was found to have a liver tumor. Plain computed tomography showed a high-density tumor that was enhanced in the arterial phase, and the enhancement was prolonged in the delayed phase. Magnetic resonance imaging showed that the tumor was hyperintense on in- and opposed-phase T1-weighted gradient recalled echo images and hyperintense on T1-weighted spin echo images. A partial hepatic resection was performed. A histological examination revealed normal hepatocyte like tumor cells, composed of abundant dark brown granules of Dubin-Johnson-like pigment, which stained positively for Masson-Fontana stain. The tumor was diagnosed to be a very rare hepatocellular adenoma with the deposition of Dubin Johnson-like pigment. PMID- 21626343 TI - Gastro-pancreaticostomy using a tube stent as a less invasive palliative treatment for malignant obstructive pancreatitis. AB - Surgical palliation of malignant obstructive pancreatitis poses a considerable burden to patients, especially those with extensive disease. We herein present a novel technique for performing gastro-pancreaticostomy using a tube stent as a less invasive palliative treatment for malignant obstructive pancreatitis. The main pancreatic duct was punctured via the wall of the pyloric antrum. After the insertion of a guidewire, a double pigtail-type tube stent with side holes was inserted into the main pancreatic duct through the wall of the pyloric antrum. This technique was useful as a means of accomplishing less invasive palliative treatment, and has the potential to improve the quality of life for patients with malignant obstructive pancreatitis. PMID- 21626344 TI - [Conservative treatment methods]. PMID- 21626345 TI - Differential immunohistochemical expression of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 in cow uteri with adenomyosis during follicular phase. AB - The present investigation was intended to show a different immunohistochemical profile of matrix metalloproteinase-2 and Tissue inhibitor metalloproteinase-2 in bovine uteri with adenomyosis during follicular phase. Uterine samples of 32 cows in reproductive age were taken from the medial third of one of the uterine horns and grouped according to the adenomyosis degree (superficial and deep). Tissue sections (4 MUm) were incubated overnight at 4 degrees C with monoclonal antibody for matrix metalloproteinase-2 and Tissue inhibitor metalloproteinase-2. Staining intensities were evaluated in the luminal epithelium, ectopic and dystopic endometrial tissue (stroma, capillaries and glands), endometrial-myometrial border, myometrium, myometrial vessels (middle tunic and endothelium). The matrix metalloproteinase-2 expression was higher for deep adenomyosis samples, showing a differential mean reactivity in superficial endometrium, myometrial vessels, myometrium adjacent to adenomyotic focus and endometrial-myometrial border (P < 0.05). Moreover, matrix metalloproteinase-2 expression was higher in deep adenomyosis samples than that of Tissue inhibitor metalloproteinase-2 in almost all uterine structures analyzed (except for the endometrial and myometrial vessels and endometrial-myometrial border). The opposite was observed in the follicular phase, for both normal specimens and with superficial adenomyosis, where Tissue inhibitor metalloproteinase-2 expression was higher than that of matrix metalloproteinase-2. In conclusion, a differential pattern of matrix metalloproteinase-2 and Tissue inhibitor metalloproteinase-2 was observed in cow uteri with adenomyosis. PMID- 21626346 TI - The fluctuation property of blown sand particles and the wind-sand flow evolution studied by numerical method. AB - Sand particles blown by wind cause serious environmental problems and many researchers are trying to understand the dynamic properties of blown sand better. But the existing numerical approaches have not been able to simulate many important characteristics of wind-sand flow. In this paper, the evolution and fluctuation properties of blown sand at a dynamic steady state are investigated by using a more effective method. Using the LES (large eddy simulation) method for air phase movement and the DEM (discrete element method) for solid phase movement along with the existing particle-bed splashing function, we have characterized the whole movement property of the wind-sand system. The results indicate that the saturation time decreases with the inlet friction velocity, and it gradually reaches the shortest saturation time of about 1s; the saturation length, which is about 14 m at the usual wind velocity, first increases with wind velocity and then reaches a plateau; within the saturation length, the sand transport rate at different positions varies with time; the sand transport rate of the stable wind-sand flow is non-uniform with distance downwind and time, and has a notable correlation with the inflow friction velocity. PMID- 21626347 TI - Allergen-specific immunotherapy: towards combination vaccines for allergic and infectious diseases. AB - IgE-mediated allergies affect more than 25% of the population. Allergen-specific immunotherapy (SIT) is an antigen-specific and disease-modifying form of treatment. It is based on the therapeutic administration of the disease-causing allergens to allergic patients. However, the fact that only allergen extracts of insufficient quality are currently available and the possible occurrence of side effects during treatment limit the broad use of SIT and prophylactic vaccination is has not yet been performed. In the last 20 years the DNA sequences of the most common allergens have been isolated and the corresponding allergens have been produced as recombinant allergens. Based on the progress made in the field of allergen characterization it is possible to improve the quality and safety of allergy vaccines and to develop new, more effective strategies for a broad application of SIT and even for prophylactic treatment. Here we discuss the development of combination vaccines for allergy and infectious diseases. This approach is based on the selection of allergen-derived peptides with reduced IgE- and T cell reactivity in order to minimize IgE- and T cell-mediated side effects as well as the potential of the vaccine to induce allergic sensitization. These peptides are fused by recombinant technology onto a viral carrier protein to obtain a combination vaccine which induces protective immunity against allergy and viral infections. The application of such combination vaccines for therapy and prophylaxis of allergy and infectious diseases is discussed. PMID- 21626348 TI - Effects of homeodomain protein CDX2 expression on the proliferation and migration of lovo colon cancer cells. AB - The homeobox gene, CDX2, plays a major role in development, especially in the gut, and also functions as a tumor suppressor in the adult colon. In the present study, we investigated the effects of CDX2 expression on the proliferation, migration, and apoptosis of the human colon cancer cell line, Lovo. Lovo cells exogenously expressing CDX2 exhibited no significant differences in the percentage of cells in G1- and S-phase or in apoptosis, as determined by flow cytometry. MTT assay also confirmed that CDX2 expression had no effect on proliferation in these cells. Interestingly, conditioned medium collected from CDX2-overexpressing Lovo cells showed a significant decrease in secretion of MMP 2 and the invasive potential of these cells was significantly inhibited. Collectively, these data suggest that CDX2 may play a critical role in the migration and metastasis of colon carcinoma and over-expression of CDX2 in colon cancer cells markedly inhibits invasion. Based on these results, exogenous expression of CDX2 might be a promising option in the treatment of colon carcinoma. PMID- 21626349 TI - Posttraumatic growth in combat veterans. AB - Combat veterans and their families face significant challenges not only to their abilities to cope, but often to their fundamental belief systems. Traumatic events represent assaults on core beliefs, yet at times, produce cognitive processing that can ultimately result in personal transformations called posttraumatic growth (PTG). Clinicians can utilize a systematic therapeutic approach to facilitate PTG as they carry out a relationship of expert companionship. PTG in service members is described in this article, as well as the approach to facilitation of PTG. PMID- 21626350 TI - Discrete and recurrent traumatization in PTSD: fear vs. anxious misery. PMID- 21626351 TI - Military medicine meets behavioral pain science. PMID- 21626352 TI - Some thoughts on trauma, pain, posttraumatic stress disorder and traumatic brain injury. PMID- 21626353 TI - Implementation of a suicide nomenclature within two VA healthcare settings. AB - Suicide and suicide attempts are significant issues for military, Veterans Affairs (VA), and civilian healthcare systems. The lack of uniform terms related to self-directed violence (SDV) has inhibited epidemiological surveillance efforts, limited the generalizability of empirical studies of suicide and non lethal forms of SDV, and complicated the implementation of evidence-based assessment and treatment strategies for individuals with suicidal thoughts and/or behaviors. The Department of Veterans Affairs recently adopted the Centers for Disease Control and Prevention's (CDC) SDV Classification System (SDVCS). This paper describes an implementation study of the SDVCS in two VA Medical Centers. The Veterans Integrated Service Network (VISN) 19 Mental Illness Research, Education and Clinical Center (MIRECC) training program for the SDVCS, including the SDVCS Clinical Tool (CT), will be discussed. Although preliminary data suggest that the CT and SDVCS are generally perceived as being acceptable and useful, further work will likely be required to facilitate widespread adoption. Potential next steps in this process are presented. PMID- 21626354 TI - Complicating factors associated with mild traumatic brain injury: impact on pain and posttraumatic stress disorder treatment. AB - The nature of combat in Iraq and Afghanistan has resulted in high rates of comorbidity among chronic pain, posttraumatic stress disorder (PTSD), and mild traumatic brain injury (mTBI) in Veterans of Operations Enduring Freedom and Iraqi Freedom (OEF/OIF). Although separate evidence-based psychological treatments have been developed for chronic pain and PTSD, far less is known about how to approach treatment when these conditions co-occur, and especially when they co-occur with mTBI. To provide the best care possible for OEF/OIF Veterans, clinicians need to have a clearer understanding of how to identify these conditions, ways in which these conditions may interact with one another, and ways in which existing evidence-based treatments can be modified to meet the needs of individuals with mTBI. The purpose of the present paper is to review the comorbidity of pain, PTSD, and mTBI in OEF/OIF Veterans, and provide recommendations to clinicians who provide care to Veterans with these conditions. First, we will begin with an overview of the presentation, symptomatology, and treatment of chronic pain and PTSD. The challenges associated with mTBI in OEF/OIF Veterans will be reported and data will be presented on the comorbidity among all three of these conditions in OEF/OIF Veterans. Second, we will present recommendations for providing psychological treatment for chronic pain and PTSD when comorbid with mTBI. Finally, the paper concludes with a discussion of the need for a multidisciplinary treatment approach, as well as a call for continued research to further refine existing treatments for these conditions. PMID- 21626355 TI - Assessment and treatment of combat-related PTSD in returning war veterans. AB - Over the past 9 years approximately 2 million U.S. military personnel have deployed in support of Operation Iraqi Freedom in Iraq and Operation Enduring Freedom in and around Afghanistan. It has been estimated that 5-17% of service members returning from these deployments are at significant risk for combat related posttraumatic stress disorder (PTSD). Many of these returning war veterans will seek medical and mental health care in academic health centers. This paper reviews the unique stressors that are related to the development of combat-related PTSD. It also reviews evidence-based approaches to the assessment and treatment of PTSD, research needed to evaluate treatments for combat-related PTSD, and opportunities and challenges for clinical psychologists working in academic health centers. PMID- 21626356 TI - The evaluation and treatment of comorbid pain and PTSD in a military setting: an overview. AB - The present article reviews the growing prevalence of comorbid pain and post traumatic stress disorder (PTSD) in the military. This has been caused by the ongoing military conflicts in Iraq and Afghanistan, where new combat conditions/strategies are causing these comorbid conditions. Fortunately, comprehensive interdisciplinary treatment programs, originally developed for a civilian population and in academic settings, are being successfully "translated" or utilized in the military environment. Recent data demonstrating this translational clinical intervention are presented. Finally, challenges encountered when translating these interventions in a military environment are also discussed. PMID- 21626357 TI - Screening, brief intervention, and referral to treatment for military spouses experiencing alcohol and substance use disorders: a literature review. AB - This paper provides an overview of alcohol and substance use issues in military spouses, and explore how the screening, brief intervention, and referral to treatment (SBIRT) model may enable health care providers to identify individuals at risk for developing substance use related disorders. The information presented is based on a broad literature scan relating to the characteristics of the military lifestyle, health infrastructure, screening and intervention processes, and the uses of SBIRT in military and civilian settings. Current literature suggests that military spouses, and families, tend to be at different points in their life course than civilian families of similar ages. Marrying earlier and having children sooner coupled with military lifestyle stressors place them at increased risk for developing adverse coping mechanisms, particularly during deployment. SBIRT has been recognized as an effective method among civilian patients although there is limited research on the efficacy of SBIRT for military spouses at risk of or experiencing substance use problems. PMID- 21626358 TI - The need for child mental health services within a department of defense setting. PMID- 21626359 TI - An education and training programme for radiological institutes: impact on the reduction of the CT radiation dose. AB - OBJECTIVES: To establish an education and training programme for the reduction of CT radiation doses and to assess this programme's efficacy. METHODS: Ten radiological institutes were counselled. The optimisation programme included a small group workshop and a lecture on radiation dose reduction strategies. The radiation dose used for five CT protocols (paranasal sinuses, brain, chest, pulmonary angiography and abdomen) was assessed using the dose-length product (DLP) before and after the optimisation programme. The mean DLP values were compared with national diagnostic reference levels (DRLs). RESULTS: The average reduction of the DLP after optimisation was 37% for the sinuses (180 vs. 113 mGycm, P < 0.001), 9% for the brain (982 vs. 896 mGycm, P < 0.05), 24% for the chest (425 vs. 322 mGycm, P < 0.05) and 42% for the pulmonary arteries (352 vs. 203 mGycm, P < 0.001). No significant change in DLP was found for abdominal CT. The post-optimisation DLP values of the sinuses, brain, chest, pulmonary arteries and abdomen were 68%, 10%, 20%, 55% and 15% below the DRL, respectively. CONCLUSIONS: The education and training programme for radiological institutes is effective in achieving a substantial reduction in CT radiation dose. PMID- 21626360 TI - The uptake transporter OATP8 expression decreases during multistep hepatocarcinogenesis: correlation with gadoxetic acid enhanced MR imaging. AB - OBJECTIVES: To clarify the changes in organic anion-transporting polypeptide 8 (OATP8) expression and enhancement ratio on gadoxetic acid-enhanced MR imaging in hepatocellular nodules during multistep hepatocarcinogenesis. METHODS: In imaging analysis, we focused on 71 surgically resected hepatocellular carcinomas (well, moderately and poorly differentiated HCCs) and 1 dysplastic nodule (DN). We examined the enhancement ratio in the hepatobiliary phase of gadoxetic acid enhanced MR imaging [(1/postcontrast T1 value-1/precontrast T1 value)/(1/precontrast T1 value)], then analysed the correlation among the enhancement ratio, tumour differentiation grade and intensity of immunohistochemical OATP8 expression. In pathological analysis, we focused on surgically resected 190 hepatocellular nodules: low-grade DNs, high-grade DNs, early HCCs, well-differentiated, moderately differentiated and poorly differentiated HCCs, including cases without gadoxetic acid-enhanced MR imaging. We evaluated the correlation between the immunohistochemical OATP8 expression and the tumour differentiation grade. RESULTS: The enhancement ratio of HCCs decreased in accordance with the decline in tumour differentiation (P < 0.0001, R = 0.28) and with the decline of OATP8 expression (P < 0.0001, R = 0.81). The immunohistochemical OATP8 expression decreased from low-grade DNs to poorly differentiated HCCs (P < 0.0001, R = 0.15). CONCLUSIONS: The immunohistochemical expression of OATP8 significantly decreases during multistep hepatocarcinogenesis, which may explain the decrease in enhancement ratio on gadoxetic acid-enhanced MR imaging. PMID- 21626361 TI - Letter to the Editor re: "Outcome analysis in 3160 implantations of radiologically guided placements of totally implantable central venous port systems": observations about right internal jugular access. PMID- 21626362 TI - Imaging features of benign solid testicular and paratesticular lesions. AB - OBJECTIVE: The presence of an intratesticular solid lesion is usually highly suspicious for malignancy. Conversely, most extratesticular solid lesions including paratesticular lesions are benign. The characteristic imaging features of malignant solid testicular lesions are well known, but various unusual causes and imaging features of benign solid testicular lesions can be particularly misleading. Therefore, a careful assessment of solid testicular and paratesticular lesions is warranted. The purpose of this article is to present the clinical and imaging features of the spectrum of benign solid testicular and paratesticular lesions. METHODS: We demonstrate a variety of benign solid testicular and paratesticular lesions and correlate them with pathologic results. RESULTS: Specific the clinical and imaging features of the spectrum of benign solid testicular and paratesticular lesions have been described. CONCLUSIONS: Familiarity with the clinical setting and imaging features of benign solid testicular and paratesticular lesions should facilitate prompt, accurate diagnosis and treatment. PMID- 21626364 TI - How small are small mutation rates? AB - We consider evolutionary game dynamics in a finite population of size N. When mutations are rare, the population is monomorphic most of the time. Occasionally a mutation arises. It can either reach fixation or go extinct. The evolutionary dynamics of the process under small mutation rates can be approximated by an embedded Markov chain on the pure states. Here we analyze how small the mutation rate should be to make the embedded Markov chain a good approximation by calculating the difference between the real stationary distribution and the approximated one. While for a coexistence game, where the best reply to any strategy is the opposite strategy, it is necessary that the mutation rate MU is less than N (-1/2)exp[-N] to ensure that the approximation is good, for all other games, it is sufficient if the mutation rate is smaller than (N ln N)(-1). Our results also hold for a wide class of imitation processes under arbitrary selection intensity. PMID- 21626365 TI - The limiting behaviour of a mainland-island metapopulation. AB - Stochastic patch occupancy models (SPOMs) are a class of discrete time Markov chains used to model the presence/absence of a population in a collection of habitat patches. This class of model is popular with ecologists due to its ability to incorporate important factors of the habitat patch network such as connectivity and distance between patches as well as heterogeneity in patch characteristics. We present an asymptotic examination of a simple type of SPOM called the mainland-island model. In this model a single patch called the mainland is connected to a large number of smaller patches called islands and each island is only connected to the mainland. We discuss the limiting behaviour of the SPOM as the number of islands increases and the size of the islands decrease relative to the mainland. We demonstrate that a variety of limiting behaviours is possible depending on the scaling of the island size and on the heterogeneity of habitat quality. PMID- 21626363 TI - Patient acceptability of CT colonography compared with double contrast barium enema: results from a multicentre randomised controlled trial of symptomatic patients. AB - OBJECTIVES: To determine patient acceptability of barium enema (BE) or CT colonography (CTC). METHODS: After ethical approval, 921 consenting patients with symptoms suggestive of colorectal cancer who had been randomly assigned and completed either BE (N = 606) or CTC (N = 315) received a questionnaire to assess experience of the clinical episode including bowel preparation, procedure and complications. Satisfaction, worry and physical discomfort were assessed using an adapted version of a validated acceptability scale. Non-parametric methods assessed differences between the randomised tests and the effect of patient characteristics. RESULTS: Patients undergoing BE were significantly less satisfied (median 61, interquartile range [IQR] 54-67 vs. median 64, IQR 56-69; p = 0.003) and experienced more physical discomfort (median 40, IQR 29-52 vs. median 35.5, IQR 25-47; p < 0.001) than those undergoing CTC. Post-test, BE patients were significantly more likely to experience 'abdominal pain/cramps' (68% vs. 57%; p = 0.007), 'soreness' (57% vs. 37%; p < 0.001), 'nausea/vomiting' (16% vs. 8%; p = 0.009), 'soiling' (31% vs. 23%; p = 0.034) and 'wind' (92% vs. 84%; p = 0.001) and in the case of 'wind' to also rate it as severe (27% vs. 15%; p < 0.001). CONCLUSION: CTC is associated with significant improvements in patient experience. These data support the case for CTC to replace BE. PMID- 21626366 TI - Management of unstable arrhythmias in cardiogenic shock. AB - OPINION STATEMENT: Atrial and ventricular arrhythmias commonly arise in the setting of cardiogenic shock and often result in hemodynamic deterioration. Causative factors include myocardial ischemia, volume overload, and metabolic disturbances. Correcting these factors plays an important role in managing arrhythmias in this setting. Ventricular arrhythmias are more ominous compared to atrial arrhythmias but both require prompt intervention with electrical shock and anti-arrhythmic drug suppression. Coronary reperfusion is key to improving survival, including reducing the risk of sudden cardiac arrest, in acute myocardial infarction. Case series have also demonstrated the value of intra aortic balloon pump counter-pulsation in suppressing ventricular arrhythmias in cardiogenic shock. The mechanism of arrhythmia suppression may be due to improved coronary perfusion and afterload reduction. Percutaneous ventricular assist device placement may be effective in this setting; however, data addressing this specific endpoint are lacking. Anti-arrhythmic drug options for ventricular and atrial arrhythmia suppression, in the setting of cardiogenic shock, are relatively limited. Common class I agents are excluded due to the inherent abnormal cardiac structure and function in the setting of cardiogenic shock. Class III drug options include dofetilide and amiodarone. The other Class III agents, sotalol and dronedarone, are excluded due to associated mortality observed in the SWORD and ANDROMEDA trials, respectively. Dofetilide is renally excreted and causes QT interval prolongation. Care should be taken to avoid excessive drug accumulation due to poor kidney perfusion and function. Dofetilide is approved for use for atrial arrhythmias and has not been studied for ventricular arrhythmia suppression. The DIAMOND-CHF trial established its safety in the setting of heart failure. Amiodarone is very effective in suppressing both atrial and ventricular arrhythmias. It is often the drug of choice in heart failure. Its off-label use for atrial arrhythmias is very common. Care should be taken with intravenous amiodarone to avoid hypotension. PMID- 21626367 TI - Distribution of microfossils within polymetallic nodules: biogenic clusters within manganese layers. AB - Biomineralization is the process by which living organisms or organic matrices produced by them initiate and structure deposition of inorganic polymers/minerals. Deep-sea polymetallic nodules and crusts have recently been recognized as biominerals that are formed around bio-seeds; these deposits are of economic value. A detailed understanding of their formation will contribute to their sustainable exploitation in the future. Polymetallic nodules grow concentrically around discrete nuclei that have recently been described as bio seeds formed from microorganisms, diatoms, or coccoliths. In the present study, polymetallic nodules from the Clarion-Clipperton Zone have been analyzed. It is described that the approximately 5-cm large polymetallic nodules are composed of micronodules (size of 100-450 MUm) that aggregated to nests (2-3 mm). High resolution scanning electron microscopy (HR-SEM) and high-resolution energy dispersive X-ray (HR-EDX) spectroscopic analyses revealed that the micronodules are composed of discrete layers of Mn and Fe. Imprints of microorganisms/microbe like assemblies are found in the Mn-rich regions of the micronodules. HR-SEM/EDX analyses confirmed that these microorganisms are surrounded by a Mn-rich environment. These findings strongly suggest that those organisms acted as bio seeds that allowed the deposition of Mn(IV) minerals which in turn helped Fe minerals to associate. Hence, these data support the concept that the growth of the polymetallic nodules starts as a biomineral and is completed by genuine mineralic depositions. It is expected that these data will contribute to the development of strategies for a sustainable exploitation of the polymetallic nodules. PMID- 21626368 TI - Secondary structure formation of homopolymeric single-stranded nucleic acids including force and loop entropy: implications for DNA hybridization. AB - Loops are essential secondary structure elements in folded DNA and RNA molecules and proliferate close to the melting transition. Using a theory for nucleic acid secondary structures that accounts for the logarithmic entropy -c ln m for a loop of length m, we study homopolymeric single-stranded nucleic acid chains under external force and varying temperature. In the thermodynamic limit of a long strand, the chain displays a phase transition between a low-temperature/low-force compact (folded) structure and a high-temperature/high-force molten (unfolded) structure. The influence of c on phase diagrams, critical exponents, melting, and force extension curves is derived analytically. For vanishing pulling force, only for the limited range of loop exponents 2 < c ? 2.479 a melting transition is possible; for c <= 2 the chain is always in the folded phase and for 2.479 ? c always in the unfolded phase. A force-induced melting transition with singular behavior is possible for all loop exponents c < 2.479 and can be observed experimentally by single-molecule force spectroscopy. These findings have implications for the hybridization or denaturation of double-stranded nucleic acids. The Poland-Scheraga model for nucleic acid duplex melting does not allow base pairing between nucleotides on the same strand in denatured regions of the double strand. If the sequence allows these intra-strand base pairs, we show that for a realistic loop exponent c ~ 2.1 pronounced secondary structures appear inside the single strands. This leads to a lower melting temperature of the duplex than predicted by the Poland-Scheraga model. Further, these secondary structures renormalize the effective loop exponent [Formula: see text], which characterizes the weight of a denatured region of the double strand, and thus affect universal aspects of the duplex melting transition. PMID- 21626369 TI - [Legionella : how well is the problem understood, how well is it controlled, and how good are the regulations?]. PMID- 21626370 TI - [Legionella in drinking water : outcomes of an exchange of ideas on the prevention of waterborne legionellosis]. AB - Although much is known about Legionella and the illness they cause, the relationship between their concentration in water and the risk of infection remains unclear. A comprehensive body of experience shows that the growth of Legionella in heated water distribution systems can be effectively controlled by following the preventive measures described in the generally acknowledged codes of practice. As orientation to trigger action, a technical action level of 100 colony-forming units in 100 ml water has found wide acceptance as the maximally tolerable concentration of Legionella in drinking water hygiene. However, this value as well as the concept for controlling the growth of Legionella in installations has developed historically. In part, the focus differs between drinking water hygiene and hospital hygiene. Also, there is no harmonized European approach for controlling Legionella. In spite of such differences, there are important international parallels in the assessment of the occurrence of Legionella, in experience with controlling them, and in recommendations for prevention. There is a need particularly for adequate studies to clarify the risk of infection as well as for the publication of existing data and experience showing the efficacy of measures for prevention, disinfection and system upgrading. Such data are necessary to support evidence-based prevention of Legionella infections and to create a better epidemiological data base in Germany. One chance for reaching this target would be to improve practices in diagnosis, reporting and central data evaluation-not only of illness, but including also data on Legionella occurrence. More frequent Legionella testing of pneumonia patients is a prerequisite for this, but particularly also for the rapid identification and removal of the source of infection. Further requirements include better training of planers, plumbers, and operators of drinking water installations about the approach to preventing Legionella contamination of drinking water installations described in standards and guidelines. The further development of practicable concepts for effective Legionella prevention requires good collaboration between public authorities responsible for drinking water and hospital hygiene on the federal and state levels and experts for the prevention of infection, hygiene, and sanitary installations. PMID- 21626371 TI - [Epidemiology of Legionnaires' disease in Germany]. AB - Legionnaires' disease (LD) is a severe form of pneumonia which is caused by bacteria of the genus Legionella. They are widespread in fresh water and can also colonize technical water systems where they might present an infection risk. Since 2001, notification of laboratory-confirmed LD is mandatory in Germany. From 2001-2009, a total of 3672 cases of LD were registered. During the first 7 years, case numbers increased (2001: 127; 2007: 536 cases) but have remained stable during the past 2 years (2008: 525; 2009: 503 cases). In 2009, 49.6% of cases were attributed to an infection in the community, 33.2% were travel associated, and 13.6% were nosocomial. The average case fatality rate between 2001 and 2009 was 6.5%. However, the case fatality rate of nosocomial cases was three times as high compared to cases with non-nosocomial exposure. The network for community acquired Legionnaires' disease (CAPNETZ) estimates that there are 20,000 cases per year in Germany. Thus, the number of reported cases represents only a small proportion of the actually occurring cases. It is likely that specific LD diagnostics are insufficiently used. Hence, physicians should test more patients with pneumonia for LD. In particular, because of the high case fatality, nosocomial pneumonia cases need to be tested; identified LD cases require rigorous investigational and corrective action. In order to obtain evidence-based data on the relationship of water contamination and the risk for LD, it would be desirable, if (in addition to the patients' epidemiological data) the results of water texts relating to a given case were also reported systematically. PMID- 21626372 TI - [Community-acquired Legionella pneumonia : data from the CAPNETZ study]. AB - Legionella are present in the environment as well as in biofilms of water installation systems. Most Legionella live in amoebae. More than 51 different species of Legionella have been identified; however, most pneumonias are caused by Legionella pneumophila serogroup 1. Legionnaire's disease has an incidence of about 4% in Germany. Most cases of Legionnaire's disease are sporadic. Microbiological identification of Legionella can be achieved by cultivation of Legionella spp. on specific media, performing of Legionella-specific PCR from respiratory samples, or Legionella urinary antigen testing. Patients with severe underlying diseases, patients receiving immunosuppression, and patients who are heavy smokers have a predisposition to Legionnaire's disease. Men are significantly more often affected. Whereas outpatients show a mild clinical course, mortality for hospitalized patients is 11.2%. It can be assumed that only a minority of cases of Legionnaire's disease is recognized and reported in Germany. PMID- 21626373 TI - [Legionella pneumophila : genetic diversity of patients and environmental isolates]. AB - Legionellae are environmental bacteria that can be frequently isolated from technical water supply systems. The most prevalent species is Legionella pneumophila, especially serogroup 1. In the environment, legionellae multiply in amoebae. Since Legionella pneumonias cannot be distinguished from pneumonias caused by other microbial pathogens, special microbiological tests, e.g., urinary antigen assays, are essential to detect Legionella infections. All water supply systems to which the patient is exposed during the incubation time of 2 to 10 days might be the source of the infection. This can be confirmed or excluded by molecular typing of isolates from patients and the environment. The most commonly used techniques are monoclonal antibody typing and sequence-based typing (SBT). Some sequence types (ST) are frequently found among clinical strains but are seldom isolated from the environment, e.g., ST 23, 42, 47, 62, and 146. It is safe to assume that such strains are highly virulent. Conversely, it does not seem to be justified to dedicate the same awareness to all environmental Legionella strains. PMID- 21626374 TI - [Experience with prevention and control of legionellosis in Germany : plea for proactive risk management]. AB - Legionellosis is meanwhile the most important specific water-associated infectious disease in developed countries, which is completely preventable, if water distribution systems are correctly planned and operated. This assumes clear criteria for risk regulation and for verification, including microbiological monitoring for Legionella. There are different reactive and proactive strategies in the USA and in Europe. The common premises for prevention and control of legionellosis in Germany, relevant facts for risk regulation, experience in Germany toward proactive risk regulation, and the current approach of the amended drinking water ordinance are discussed. The article concludes with a short discussion of the controversial approaches for the prevention of legionellosis in Germany. PMID- 21626375 TI - [Practical and legal aspects on the presence of Legionella in drinking water installations]. AB - A drinking water installation is often only a small component of building engineering systems and in general practice it is rarely the focus of the work of local personnel. There are not only the specifications of manufacturers, but also numerous "generally accepted rules of technology" must be complied with when operating these building engineering systems. The foremost ones pertinent to this topic are discussed in this article. However, noncompliance with the "generally accepted rules of technology" can cause contamination with Legionella, and the actions that would then need to be taken must be based on each specific case. The prerequisite for permanently eliminating any contamination with Legionella is identifying the precise causes and basing all ensuing action on that. Correctly transforming the existing "generally accepted rules of technology" into reality would be a sufficient guarantee to reach a target of less than 100 cfu/100 ml for Legionella. This article illustrates that this target can be reached by renovation and without applying any disinfectant processes even in drinking water installations with an extremely high level of contamination. Furthermore, this article presents examples of the legal aspects of the topic of Legionella from the point of view of the engineer. Finally, the trend toward operating drinking water installations at a lower temperature to save energy while applying disinfection is viewed critically by the authors. PMID- 21626376 TI - [Generally accepted engineering standards and Legionella in drinking water : findings from Frankfurt am Main]. AB - To investigate the association between the engineering standards of drinking water systems and the extent of the Legionella colonization, we subjected our 2006 published data of the drinking water monitoring of the city's public health services for Frankfurt on Main, Germany, to closer analysis. A total of 413 records were available, with both technical data and results of the Legionella colonization. When comparing the classes of technical deficiencies of the drinking water installations with the Legionella colonization results, 93% of the Legionella data "not dedectable in 100 ml" were found in the group of drinking water installations that were technically assessed as "free of deficiencies". Thus, "good" technical engineering is associated with low or lack of Legionella colonization with a high probability even with hints for a dose-effect phenomenon the more engineering deficiencies that exist, the higher the Legionella contamination. PMID- 21626377 TI - Legionellosis prevention in France. AB - Since 1997, both the prevention of legionellosis and the control of Legionella in water networks and cooling towers have greatly improved in France. The epidemiological surveillance of Legionnaires' disease showed an increase, which was followed by a decrease in the incidence after a maximum was reached in 2005 (incidence rate of 2.5 per 10(5)). A steady decrease in the incidence rate has been observed since 2006, mainly due to the efficacy of control measures both for cooling towers and hospital water networks. In hospitals, a proactive approach was proposed in 1998 and quantitative limit values with systematic survey were put into force in 2002. The percentage of nosocomial cases has decreased dramatically and is now around 6-7% of all cases recorded in France. No nosocomial outbreak has been reported in recent years. However, this is not the case for community cases linked to diverse exposure sources, including cooling towers. PMID- 21626378 TI - [Campylobacteriosis of man : livestock as reservoir for Campylobacter species]. AB - Over the last few years, infections with Campylobacter have significantly increased in Europe and Germany and these bacteria have even surpassed Salmonella as the most prevalent bacteria, causing gastroenteritis. Especially contamination during the handling and consumption of meat products seems to be the most important risk factor which plays a prominent role for transmission to man. In addition, contact with pets and other animals, drinking raw or improperly pasteurized milk, and the tenacity of Campylobacter in different environments, especially water, have also to be considered for an adequate risk assessment. Besides gastroenteritis, arthralgia, and Guillain-Barre syndrome are important clinical complications of Campylobacter infections in man. At the same time, it is mostly unclear why the course of infection in man and in reservoir animals differs significantly, especially as only a few classical bacterial virulence factors have been identified so far. For these reasons, the development of efficient prevention strategies is of utmost importance in order to control campylobacteriosis. PMID- 21626379 TI - [Social living conditions and health among mothers in Germany : findings from a population sample]. AB - This paper reports findings from a population sample of mothers with underage children living in Germany (n=3,129). The objective of the study was to analyze whether social and familiar living conditions are associated with enhanced health risks for mothers. The sample is representative with respect to German federal states, school education, marital status, age of mothers, and number of children. Health problems were assessed in terms of physical disabilities and discomforts, anxiety and depression, and self-rated health. About 27% of mothers perceived physical disabilities and discomfort, 21% reported high levels of anxiety, and 22% high levels of depression. About 6% assessed their health as poor or very poor. Particularly single motherhood, unemployment, sole responsibility for household and family as well as low income and low school education increased the risk of poor health. The findings suggest that for specific subgroups of mothers parenting may be a vulnerable phase of life reflecting the need for a life stage orientation in health inequality research. PMID- 21626380 TI - [Monitoring and quality assurance of prevention and health promotion at the federal level]. AB - Monitoring and quality assurance are gaining in importance for the identification of needs and the effectiveness of prevention and health promotion activities. This paper presents examples of activities of monitoring and quality assurance at the federal level, carried out by the Federal Centre for Health Education and the Robert Koch Institute. Examples include the prevention issues "HIV/AIDS", "nutrition and physical activity" and "child health". They illustrate the roles of epidemiological surveillance, health monitoring, evaluation, and intervention reporting. The Robert Koch Institute and the Federal Centre for Health Education provide complementary information on health and intervention reporting at the federal level. With their reports, they provide essential information for health policy to formulate, to implement and to evaluate evidence-based national health goals and action plans. PMID- 21626381 TI - [Self-assessment of BMI data : verification of the practicability of a correction formula on a sample of 11- to 13-year-old girls]. AB - The decision to measure or to ask about data concerning height and weight in order to calculate body mass index (BMI) has an influence on the economy and validity of the measurements. Although self-reported information is less expensive, this information may possibly have a bias on the determined prevalences of different weight groups. Using representative data from the KiGGS study with a comparison of directly measured and self-reported BMI data, Kurth and Ellert (2010) developed two correction formulas for prevalences resulting from self-reported information. The aim of the study was to examine the practicability of the proposed correction formulas on our own data concerning self-reported BMI data of 11- to 13-year-old girls (n=1,271) and to assess the plausibility of the corrected measurements. As a result, the prevalences of our own data changed in the expected direction both for underweight and for overweight. Both formulas were found to be practicable, the consideration of the subjective weight status (formula 2) resulted in a greater change in prevalences compared to the first correction formula. PMID- 21626382 TI - [Report of the Central Ethics Committee for Stem Cell Research (ZES) : eighth report after the enactment of the Stem Cell Act (StZG), Reporting Period: 1 December 2009 to 31 December 2010]. PMID- 21626383 TI - [15th Notification of changes to the list of substances and disinfection treatment process according to AS 11 Drinking Water Ordinance 2,001 June 2011]. PMID- 21626384 TI - [Substance monograph for phthalates - New and updated reference and oxidized monoester metabolites in urine of children-and adults. Opinion of the Commission "Human-Biomonitoring" by the Federal Environment Agency]. PMID- 21626385 TI - [Classification of water-polluting substances]. PMID- 21626386 TI - Epidemiology and etiology of Parkinson's disease: a review of the evidence. AB - The etiology of Parkinson's disease (PD) is not well understood but likely to involve both genetic and environmental factors. Incidence and prevalence estimates vary to a large extent-at least partly due to methodological differences between studies-but are consistently higher in men than in women. Several genes that cause familial as well as sporadic PD have been identified and familial aggregation studies support a genetic component. Despite a vast literature on lifestyle and environmental possible risk or protection factors, consistent findings are few. There is compelling evidence for protective effects of smoking and coffee, but the biologic mechanisms for these possibly causal relations are poorly understood. Uric acid also seems to be associated with lower PD risk. Evidence that one or several pesticides increase PD risk is suggestive but further research is needed to identify specific compounds that may play a causal role. Evidence is limited on the role of metals, other chemicals and magnetic fields. Important methodological limitations include crude classification of exposure, low frequency and intensity of exposure, inadequate sample size, potential for confounding, retrospective study designs and lack of consistent diagnostic criteria for PD. Studies that assessed possible shared etiological components between PD and other diseases show that REM sleep behavior disorder and mental illness increase PD risk and that PD patients have lower cancer risk, but methodological concerns exist. Future epidemiologic studies of PD should be large, include detailed quantifications of exposure, and collect information on environmental exposures as well as genetic polymorphisms. PMID- 21626387 TI - [Hypothermia under olanzapine treatment: clinical case series and review of current literature]. AB - BACKGROUND: Antipsychotic drugs may lead to hypothermia as well as hyperthermia. Although known for decades and clinically highly relevant, the mechanisms by which antipsychotic drugs alter thermoregulatory processes in the human body are still far from being fully understood. In clinical practice, much attention is paid to antipsychotic drug-induced elevation of body core temperature as observed in the neuroleptic malignant syndrome (NMS). But also hypothermia is a clinically highly relevant adverse reaction to antipsychotic drugs. MATERIAL AND METHODS: Here we report a case series of three patients who developed severe hypothermia after administration of olanzapine. A review of the current literature is given with a focus on risk factors for the development of antipsychotic drug-induced hypothermia and its pathophysiologic mechanisms. RESULTS: A 51-year-old female patient suffering from catatonic schizophrenia, cachectic nutritional condition and hypothyroidism developed severe hypothermia of 30.0 degrees C body core temperature after administration of 30 mg olanzapine per day under comedication with lorazepam and L-thyroxine. A 48-year-old female patient with catatonic schizophrenia showed hypothermia of 31.0 degrees C (rectal measurement) after single-dose administration of olanzapine 10 mg orally and a total of 3 mg lorazepam (1-1-1 mg). The third case report describes a 69-year-old male patient with acute delusional disorder exhibiting hypothermia of 33.0 degrees C (rectal measurement) in combination with a reversible atrioventricular block grade III without any further comedication. CONCLUSION: A review of the current literature reveals that thermoregulatory disturbances as sequelae of antipsychotic drug administration depend on individual disposition as well as various independent risk factors such as environmental temperature, somatic comorbidities, endocrinological abnormalities (e.g. hypothyroidism) and structural damage of the brain. A complex interaction of dopaminergic regulatory mechanisms in the ventral hypothalamus and peripheral vaso- and sudomotor adjustments seems to be causative. Hypothermia following antipsychotic drug administration represents a serious adverse drug reaction and a potentially life-threatening event. PMID- 21626388 TI - [Socio-medical aspects of mental disorders: mental disorders in the German medical health care system and socio-medical therapeutic interventions]. AB - In the first article social medicine was defined. In this second article the health care system and special socio-medical treatments for the individual are described. The role of social medicine is to describe and optimize the health care system. The legal basis for the structure of the health care system is the Social Law. The individual encounter between therapists and patients is also regulated by law. This includes a contract of personal service. Liability of therapists is therefore restricted to correct treatment, which must be documented, and not depending on outcome. Therapists must cooperate with each other. There are special socio-medical treatments for individuals. This includes sickness certificates, enforced treatment, guardianship, support by counselling services, socio-medical support by physicians and psychotherapists in private practice, care and assessment in rehabilitation centres, and measures for reintegration into working life. PMID- 21626389 TI - Comment on Cheng et al.: Comparison study of two surgical options for distal tibia fracture-minimally invasive plate osteosynthesis vs. open reduction and internal fixation. PMID- 21626390 TI - Reconstruction of the patella with an autogenous iliac graft: clinical and radiologic results in thirteen patients. AB - PURPOSE: Extension lag, quadriceps weakness and subluxation of the extensor apparatus are known complications of patellectomy. In the case of total knee joint replacement with a nonconstrained system an instability may be encountered. Reconstruction of the patella allows restoration of the moment arm to improve quadriceps leverage. The goal of our study was to analyse the clinical and radiological results after reconstruction of the patella with an autogenous iliac graft. METHODS: 13 previously patellectomized patients had reconstruction of the patella with an autogenous iliac graft and were retrospectively studied by clinical and radiographic examination. For evaluation we used the scores of Feller and the Knee Society. Also, all complications were recorded. RESULTS: After an average follow-up of 40.1 months, nine patients had full strength of the quadriceps, while six had an improved function of the extensor apparatus. The mean Feller score was 21.8 and the Knee Society score was 67.3 for knee and 57.5 for function. Six complications occurred including three infections, two problems with the replaced patella and one fracture of the anterior superior iliac spine. CONCLUSIONS: Reconstruction of the patella with an autogenous iliac graft enables the strength of the extensor apparatus with restoration of the knee joint. PMID- 21626391 TI - Minimally invasive stabilisation of posterior pelvic-ring instabilities with a transiliac locked compression plate. AB - PURPOSE: Sacroiliac screw fixation is the method of choice for the definitive treatment of unstable posterior pelvic-ring injuries; however, this technique is demanding and associated with a high risk of iatrogenic neurovascular damage. This study evaluates whether minimally invasive transiliac locked compression plate stabilisation may be an alternative to sacroiliac screw fixation in unstable posterior pelvic-ring injuries. METHODS: We performed a retrospective analysis of patients with unstable pelvic-ring injuries treated with a transiliac locked compression plate at a level I trauma centre. Outcome evaluation was assessed using the Pelvic Outcome Score and analysis of complications, intraoperative fluoroscopic time, and duration of the surgical procedure. RESULTS: Twenty-one patients were available for follow-up after an average of 30 months. The main findings were as follows: Overall outcome for the Pelvic Outcome Score was excellent in 47.6% (ten patients), good in 19% (four patients), fair in 28.6% (six patients), and poor in 4.8% (one patient). Average operation time was 101 min and intraoperative fluoroscopic time averaged 74.2 s. No iatrogenic neurovascular injuries were observed. CONCLUSION: Minimally invasive transiliac locked compression plate stabilisation may be a good alternative to sacral screw fixation because it is quick, safe and associated with a good functional outcome. PMID- 21626392 TI - In search of the causes of Parkinson's disease, seasons 1 to 4. PMID- 21626394 TI - A multiscale mechanobiological modelling framework using agent-based models and finite element analysis: application to vascular tissue engineering. AB - Computational models of mechanobiological systems have been widely used to provide insight into these systems and also to predict their behaviour. In this context, vascular tissue engineering benefits from further attention given the challenges involved in developing functional low calibre vascular grafts with long-term patency. In this study, a novel multiscale mechanobiological modelling framework is presented, which takes advantage of lattice-free agent-based models coupled with the finite element method to investigate the dynamics of VSMC growth in vascular tissue engineering scaffolds. The results illustrate the ability of the mechanobiological modelling approach to capture complex multiscale mechanobiological phenomena. Specifically, the framework enabled the study of the influence of scaffold compliance and loading regime in regulating the growth of VSMCs in vascular scaffolds and their role in development of intimal hyperplasia (IH). The model demonstrates that low scaffold compliance compared to host arteries leads to increased luminal ingrowth and IH development. In addition, culture of a tissue-engineered blood vessel under a pulsatile luminal pressure reduced luminal ingrowth and enhanced collagen synthesis within the scaffold compared to non-pulsatile culture. The mechanobiological framework presented provides a robust platform for testing hypotheses in vascular tissue engineering and lends itself to use as an optimisation design tool. PMID- 21626393 TI - Association between the STin2 VNTR polymorphism and smoking behavior in oral cancer patients and healthy individuals. AB - The serotonergic system may be involved in smoking behavior since the intake of nicotine increases serotonin secretion in the CNS. Moreover, evidence supporting the beneficial effect of selective serotonin reuptake for quitting smoking suggesting that the serotonin transporter (5-HTT) is a plausible target for the understanding and elucidation of smoking behavior. The transcriptional activity of its human gene (SLC6A4) is modulated by a polymorphism described in the second intron, the STin2 VNTR, which thus may interfere with 5-HTT synthesis. In this study was analyzed the polymorphism STin2 VNTR of 60 smokers male patients diagnosed for oral carcinoma, 61 male smokers without cancer and 65 non-smoker healthy blood donors. The STin2. 9 allele carriers were more present in smoker groups (with cancer and without cancer, respectively) than in the non-smoker (OR = 7.11, 95% CI = 0.83-60.91 and OR = 24.73; IC 95% = 3.17-192.66). Conversely, individuals carrying allele 10 were more prevalent in non-smokers compared with smokers (oral cancer patients and individuals without cancer, respectively), showing a protective factor of this allele (OR = 0.56; 95% CI = 0.24-1.33 and OR = 0.46; 95% CI = 0.20-1.07). This is the first report of a study assessing the importance of STin2 VNTR smoking behavior in Brazilian individuals and the association of STin2. 9 allele carriers in nicotine dependence. It is suggested that individuals with low serotonin concentration in the central nervous system, probably due to the presence of the allele for high expression of 5 HTT,especially STin2. 9, were more susceptible to nicotine dependence. Moreover, individuals with the 10 allele might have less risk for nicotine dependence. PMID- 21626395 TI - Design and analysis of a head-mounted parallel kinematic device for skull surgery. AB - PURPOSE: Precision skull surgery requires specialized instrumentation to satisfy demanding requirements in cochlear array implantation, deep brain stimulation electrode placement, and related applications. A miniaturized reconfigurable parallel kinematic mechanism which can be directly mounted on a patient's skull was designed, built, and tested for precision skull surgery. METHODS: A Stewart Gough platform is attached to a patient's skull so no optical tracking affecting the overall accuracy in keyhole surgery is required. Six bone anchors comprising the mechanism base joints are implanted at positions with sufficient skull thickness. Since no fixation frame is required, intervention planning flexibility is increased. The centers of the spherical shaped bone anchors can be localized accurately in the image space. An implicit registration to the physical space is performed by assembling the kinematics. Registration error is minimized compared to common optical tracker-based approaches. Due to the reconfigurable mechanism, an optimization of the hexapod's configuration is needed to maximize accuracy and mechanical stability during the incision. Mathematical simulation was conducted to estimate system performance. RESULTS: Simulation results revealed significant improvement in accuracy and stability when exploiting the redundant degrees of freedom and the implemented reconfigurability. Inaccurate localization of base points in the image data set was identified as the main source of error. A first prototype with passive prismatic actuators, i.e. micrometer calipers, was successfully built. CONCLUSIONS: A head-mounted parallel kinematic device for high precision skull surgery was developed that provides submillimetric accuracy in straight-line incisions. The system offers enhanced flexibility due to the absence of a rigid fixation frame. PMID- 21626396 TI - MITK-ToF--range data within MITK. AB - PURPOSE: The time-of-flight (ToF) technique is an emerging technique for rapidly acquiring distance information and is becoming increasingly popular for intra operative surface acquisition. Using the ToF technique as an intra-operative imaging modality requires seamless integration into the clinical workflow. We thus aim to integrate ToF support in an existing framework for medical image processing. METHODS: MITK-ToF was implemented as an extension of the open-source C++ Medical Imaging Interaction Toolkit (MITK) and provides the basic functionality needed for rapid prototyping and development of image-guided therapy (IGT) applications that utilize range data for intra-operative surface acquisition. This framework was designed with a module-based architecture separating the hardware-dependent image acquisition task from the processing of the range data. RESULTS: The first version of MITK-ToF has been released as an open-source toolkit and supports several ToF cameras and basic processing algorithms. The toolkit, a sample application, and a tutorial are available from http://mitk.org. CONCLUSIONS: With the increased popularity of time-of-flight cameras for intra-operative surface acquisition, integration of range data supports into medical image processing toolkits such as MITK is a necessary step. Handling acquisition of range data from different cameras and processing of the data requires the establishment and use of software design principles that emphasize flexibility, extendibility, robustness, performance, and portability. The open-source toolkit MITK-ToF satisfies these requirements for the image guided therapy community and was already used in several research projects. PMID- 21626397 TI - cGMP modulates responses to queen mandibular pheromone in worker honey bees. AB - Responses to social cues, such as pheromones, can be modified by genotype, physiology, or environmental context. Honey bee queens produce a pheromone (queen mandibular pheromone; QMP) which regulates aspects of worker bee behavior and physiology. Forager bees are less responsive to QMP than young bees engaged in brood care, suggesting that physiological changes associated with behavioral maturation modulate response to this pheromone. Since 3',5'-cyclic guanosine monophosphate (cGMP) is a major regulator of behavioral maturation in workers, we examined its role in modulating worker responses to QMP. Treatment with a cGMP analog resulted in significant reductions in both behavioral and physiological responses to QMP in young caged workers. Treatment significantly reduced attraction to QMP and inhibited the QMP-mediated increase in vitellogenin RNA levels in the fat bodies of worker bees. Genome-wide analysis of brain gene expression patterns demonstrated that cGMP has a larger effect on expression levels than QMP, and that QMP has specific effects in the presence of cGMP, suggesting that some responses to QMP may be dependent on an individual bees' physiological state. Our data suggest that cGMP-mediated processes play a role in modulating responses to QMP in honey bees at the behavioral, physiological, and molecular levels. PMID- 21626398 TI - Managing repeat digital radiography images-a systematic approach and improvement. AB - Repeat analysis is an important issue for improving image quality in the field of radiology. However, the required data for repeat analysis is not easy to collect, and the accuracy of the analysis results remains controversial. The main purpose of this study is to introduce a systematic approach and, with the assistance of information technology, to improve the accuracy of data collection methods and repeat analysis in a fully digital environment. Another purpose of the study was to reduce the human resources required to maintain image quality on a daily basis. The main participant in this study is the radiology department of a medical center in Taiwan. The hospital had previously implemented a Picture Archiving and Communication Systems (PACS), which was seamlessly integrated with Radiology Information Systems (RIS) and Hospital Information Systems (HIS). A comprehensive mechanism was built for repeat analysis. The analysis was primarily achieved through comparing the difference between the amount of accumulated digital radiography (DR) images and uploaded PACS images with data mining tools. Initially, the radiologic technologists seemed to be resistant to the new quality assurance mechanism, which introduce inaccuracy into the collected data. However, after introducing the improved standard operating procedures with the proposed approach for radiologic technologists, the number of DR images generated became optimal for comparison with the number of PACS images, which made this mechanism feasible. Furthermore, information was collected regarding the reasons for repeat images and was used for improving image quality. The results revealed that the new mechanism was both effective and accurate in the analysis of repeat images. PMID- 21626399 TI - Efficient health information management systems using wireless communications technology to aid disaster victims. AB - Japan is an earthquake-prone country, and disasters have a devastating effect on the lives of residents in stricken areas. Shelters can be constructed in order to secure the physical safety of residents, but there are no such provisions for the shock of experiencing a disaster, losing property and friends, and transitioning to an unfamiliar life in a shelter, all of which can lead to mental disorders. Caretakers such as medical doctors and nurses who are dispatched to disaster sites also face difficulties in the disruption of communications and transportation, thus a system able to secure efficient health management in those facilities is also required. This paper proposes a health information management system that utilizes mobile phone cameras and mark-sensing cards to improve recovery conditions in disaster-stricken areas. PMID- 21626400 TI - Vitamin A and lipid metabolism: relationship between hepatic stellate cells (HSCs) and adipocytes. AB - Vitamin A or retinol plays a major role in the regulation of cellular homeostasis. Retinyl palmitate remains the main chemical form of vitamin A storage and is mainly located in hepatic stellate cells (HSCs) in lipid droplets resembling those found in adipose cells. White adipose tissue (WAT), is essentially involved in the regulation of lipid metabolism, through its role in lipid storage, and might also be considered as a vitamin A storage and metabolism site. WAT contains all the intracellular equipment for vitamin A metabolism and signaling pathways which allows retinol to be metabolized into retinoic acid, known to control genomic expression in WAT. The description of molecular mechanisms involved in the activation of HSCs and the differentiation of preadipocytes reveal similar cellular and molecular mechanisms. Indeed HSCs and adipocytes share a common expression of key transcription factors like PPAR-gamma and RXR known to influence perilipin expression, which play fundamental roles in lipid droplet metabolism. Both cells are also sources of important endocrine signaling secretions influencing the expression of these transcription factors. The morphological and functional characteristics of HSCs and adipocytes, including the metabolism of vitamin A and other lipids and their related signaling pathways, are summarized and compared in this review. We highlight the complexity of the interrelationship between lipids and vitamin A metabolism and the role of the complex communication existing between HSCs and WAT in diseases such as non-alcoholic fatty liver disease which is the hepatic manifestation of the metabolic syndrome. PMID- 21626402 TI - Dual effects of beta3 integrin subunit expression on human pancreatic cancer models. AB - BACKGROUND: Pancreatic cancer, the fifth leading cause of adult cancer death in Western countries, lacks early detection, and displays significant dissemination ability. Accumulating evidence shows that integrin-mediated cell attachment to the extracellular matrix induces phenotypes and signaling pathways that regulate tumor cell growth and migration. METHODS: In view of these findings, we examined the role of beta(3) in pancreatic cancer by generating two stable beta(3) expressing pancreatic human cell lines and characterizing their behavior in vitro and in vivo. RESULTS: Transduction of beta(3) selectively augmented the functional membrane alpha(v)beta(3) integrin levels, as evident from the enhanced adhesion and migration abilities related to active Rho GTPases. No effects on in vitro anchorage-dependent growth, but higher anoikis were detected in beta(3) overexpressing cells. Moreover, tumors expressing beta(3) displayed reduced growth. Interestingly, treatment of mice with an alpha(v)-blocking antibody inhibited the growth of beta(3)-expressing tumors to a higher extent. CONCLUSIONS: Our results collectively support the hypothesis that alpha(v)beta(3) integrin has dual actions depending on the cell environment, and provide additional evidence on the role of integrins in pancreatic cancer, which should eventually aid in improving prediction of the effects of therapies addressed to modulate integrin activities in these tumors. PMID- 21626403 TI - Rationale and design of the SPICE study-septal positioning of ventricular ICD electrodes. AB - AIM: Detrimental effects of right ventricular apical pacing have prompted the search for alternative pacing sites. Data from pacemaker studies suggest that the mid-septum may be a hemodynamically favorable pacing site within the right ventricle. However, data on the safety of implantable cardioverter defibrillator (ICD) lead placement in this location are limited. The aim of the SPICE study is to ascertain that implantation of a ventricular ICD lead at the mid-septum is not inferior to the traditional apical site in terms of feasibility and safety. METHODS: In total, 286 patients with the indication for ICD implantation will be included in the study and stratified in a 1:1 ratio to have the lead placed in the right ventricular apex or the mid-septum. Primary outcome measure of this randomized, prospective study is the event-free survival of lead-related complications at 3 months. The combined end point includes the feasibility to reach the randomized pacing site in combination with the achievement of adequate pacing, sensing, and defibrillation thresholds. Data analysis and sample size calculation are based on a non-inferiority design. CONCLUSION: This trial will primarily assess the feasibility, safety, and efficacy of right ventricular ICD lead positioning in the mid-septum compared to the traditional apical position. Documentation of non-inferiority of the novel lead position in terms of applicability and safety will be the prerequisite for potential further trials that may evaluate long-term clinical effects associated with mid-septal lead positions. PMID- 21626404 TI - Adsorption of ribose nucleotides on manganese oxides with varied mn/o ratio: implications for chemical evolution. AB - Manganese exists in different oxidation states under different environmental conditions with respect to redox potential. Various forms of manganese oxides, namely, Manganosite (MnO), Bixbyite (Mn(2)O(3)), Hausmannite (Mn(3)O(4)) and Pyrolusite (MnO(2)) were synthesized and their possible role in chemical evolution studied. Adsorption studies of ribose nucleotides (5'-AMP, 5'-GMP, 5' CMP and 5'-UMP) on these manganese oxides at neutral pH, revealed a higher binding affinity to manganosite (MnO) compared to the other manganese oxides. That manganese oxides having a lower Mn-O ratio show higher binding affinity for the ribonucleotides indirectly implies that such oxides may have provided a surface onto which biomonomers could have been concentrated through selective adsorption. Purine nucleotides were adsorbed to a greater extent compared to the pyrimidine nucleotides. Adsorption data followed Langmuir adsorption isotherms, and X( m ) and K( L ) values were calculated. The nature of the interaction and mechanism was elucidated by infrared spectral studies conducted on the metal oxide and ribonucleotide-metal-oxide adducts. PMID- 21626405 TI - Metabolomic analysis of sulfur-containing substances and polyamines in regenerating rat liver. AB - We studied the significance of alterations in the metabolomics of sulfur containing substances in rapidly regenerating rat livers. Male rats were subjected to two-thirds partial hepatectomy (PHx), and the changes in hepatic levels of major sulfur-containing amino acids and related substances were monitored for 2 weeks. Liver weight began to increase from 24 h after the surgery, and appeared to recover fully in 2 weeks. Serum alanine aminotransferase and aspartate aminotransferase activities were elevated immediately after the surgery and returned slowly to normal levels in 2 weeks. Methionine, S adenosylmethionine (SAM), cystathionine and cysteine were increased rapidly and remained elevated for longer than 1 week. Hepatic glutathione concentration was increased gradually for 24 h, and then decreased thereafter, whereas hypotaurine was elevated drastically right after the surgery. Hepatic concentrations of polyamines were altered significantly by PHx. In the hepatectomized livers putrescine concentration was elevated rapidly, reaching a level 40- to 50-fold greater than normal in 6-12 h. Ornithine, the metabolic substrate for putrescine synthesis, was also elevated markedly. Spermidine was increased significantly, whereas spermine was depressed below normal, which appeared to be due to the increased consumption of decarboxylated SAM for spermidine biosynthesis. The results show that the metabolomics of sulfur-containing amino acids and related substances is altered profoundly in regenerating rat livers until the original weight is recovered. Hepatic concentrations of polyamines after PHx are closely associated with the alteration in the metabolomics of sulfur-containing substances. The implication of these changes in the progression of liver regeneration is discussed. PMID- 21626406 TI - Lack of relationship between sedentary behaviour and vascular function in children. AB - Some evidence suggests that sedentary behaviour is independently associated with cardiovascular (CV) risk. Endothelial dysfunction is the earliest detectable manifestation of CVD and a strong independent predictor of CV events. No previous study has examined the relationship between sedentary behaviour and endothelial function. We assessed the basal association between conduit artery endothelial function and sedentary behaviour in children, along with the correlation between changes in sedentary behaviour and endothelial function. We studied 116 children (70?: 10.7 +/- 0.3; 46?: 10.7 +/- 0.3 years) on two occasions; in the summer (June) and late autumn (November). We assessed endothelial function via flow mediated dilation (FMD) using high-resolution Doppler ultrasound. Sedentary behaviour (SB) was assessed using objective uni-axial accelerometry. At baseline, there were no significant differences between girls and boys for any measured variables with the exception of total physical activity time. FMD was not associated with sedentary behaviour in either group or in the cohort as a whole. Although FMD decreased (10.0 +/- 4.3-7.9 +/- 3.9%, P < 0.001) and SB increased (499.1 +/- 103.5-559 +/- 81.6 min/day, P < 0.001) between the seasons, no relationship existed between changes in these variables. Our data suggest that sedentary behaviour and changes in sedentary behaviour are not associated with endothelial function in children. PMID- 21626407 TI - In vivo assessment of antiemetic drugs and mechanism of lycorine-induced nausea and emesis. AB - Lycorine is the main alkaloid of many Amaryllidaceae and known to cause poisoning with still unknown mechanisms. Longer lasting toxicological core symptoms of nausea and emesis may become a burden for human and animal patients and may result in substantial loss of water and electrolytes. To optimise the only empirical symptomatic antiemetic drug treatment at present, it is important to elucidate the causative involved targets of lycorine-induced emesis. Therefore, in the current study, we have tested the actions of a various antiemetic drugs with selective receptor affinities on lycorine-induced nausea and emesis in vivo in dogs. Beagle dogs were pre-treated in a saline vehicle-controlled crossover and random design with diphenhydramine, maropitant, metoclopramide, ondansetron or scopolamine prior lycorine administration (2 mg/kg subcutaneously). In vivo effects were assessed by a scoring system for nausea and emesis as well as by the number and lag time of emetic events for at least 3 h. Moreover, plasma pharmacokinetic analysis was carried out for ondansetron before and after lycorine injection. The data show that histaminergic (H1), muscarinic and dopaminergic (D2) receptors are presumably not involved in lycorine-induced emetic effects. While ondansetron significantly reduced the number of emetic events, lycorine-induced emesis was completely blocked by maropitant. Only ondansetron also significantly decreased the level of nausea and was able to prolong the lag time until onset of emesis suggesting a preferential participation of 5-HT3 receptors in lycorine-induced nausea. Thus, it is the first in vivo report evidencing that predominantly neurokinin-1 (NK1) and to a lesser extent 5-hydroxytryptamine 3 (5-HT3) receptors are involved in lycorine induced emesis facilitating a target-oriented therapy. PMID- 21626408 TI - Human body temperature and new approaches to constructing temperature-sensitive bacterial vaccines. AB - Many of the live human and animal vaccines that are currently in use are attenuated by virtue of their temperature-sensitive (TS) replication. These vaccines are able to function because they can take advantage of sites in mammalian bodies that are cooler than the core temperature, where TS vaccines fail to replicate. In this article, we discuss the distribution of temperature in the human body, and relate how the temperature differential can be exploited for designing and using TS vaccines. We also examine how one of the coolest organs of the body, the skin, contains antigen-processing cells that can be targeted to provoke the desired immune response from a TS vaccine. We describe traditional approaches to making TS vaccines, and highlight new information and technologies that are being used to create a new generation of engineered TS vaccines. We pay particular attention to the recently described technology of substituting essential genes from Arctic bacteria for their homologues in mammalian pathogens as a way of creating TS vaccines. PMID- 21626409 TI - Breaking T cell tolerance to beta cell antigens by merocytic dendritic cells. AB - In type 1 diabetes (T1D), a break in central and peripheral tolerance results in antigen-specific T cells destroying insulin-producing, pancreatic beta cells. Herein, we discuss the critical sub-population of dendritic cells responsible for mediating both the cross-presentation of islet antigen to CD8(+) T cells and the direct presentation of beta cell antigen to CD4(+) T cells. These cells, termed merocytic dendritic cells (mcDC), are more numerous in non-obese diabetic (NOD), and antigen-loaded mcDC rescue CD8(+) T cells from peripheral anergy and deletion, and stimulate islet-reactive CD4(+) T cells. When purified from the pancreatic lymph nodes of overtly diabetic NOD mice, mcDC can break peripheral T cell tolerance to beta cell antigens in vivo and induce rapid onset T cell mediated T1D in young NOD mouse. Thus, the mcDC subset appears to represent the long-sought critical antigen-presenting cell responsible for breaking peripheral tolerance to beta cell antigen in vivo. PMID- 21626410 TI - Substantia nigra echogenicity in Parkinson's disease: relation to serum iron and C-reactive protein. AB - In Parkinson's disease (PD), substantia nigra hyperechogenicity (SN-h) has been related to both, local iron accumulation and microglia activation. We analysed its relationship in PD patients with serum iron (n = 31) and C-reactive protein (CRP; n = 193). SN-h correlated with lower CRP and iron levels. Also, patients with a first-degree relative with PD had lower iron levels. Microglia activation, if reflected by SN-h, may be therefore unrelated to serum CRP. Findings support the idea that SN-h indicates inherited alteration of iron metabolism. PMID- 21626411 TI - Set shifting and working memory in adults with attention-deficit/hyperactivity disorder. AB - Compared to the high number of studies that investigated executive functions (EF) in children with attention-deficit/hyperactivity disorder (ADHD), a little is known about the EF performance of adults with ADHD. This study compared 37 adults with ADHD (ADHD(total)) and 32 control participants who were equivalent in age, intelligence quotient (IQ), sex, and years of education, in two domains of EF- set shifting and working memory. Additionally, the ADHD(total) group was subdivided into two subgroups: ADHD patients without comorbidity (ADHD(-), n = 19) and patients with at least one comorbid disorder (ADHD(+), n = 18). Participants fulfilled two measures for set shifting (i.e., the trail making test, TMT and a computerized card sorting test, CKV) and one measure for working memory (i.e., digit span test, DS). Compared to the control group the ADHD(total) group displayed deficits in set shifting and working memory. The differences between the groups were of medium-to-large effect size (TMT: d = 0.48; DS: d = 0.51; CKV: d = 0.74). The subgroup comparison of the ADHD(+) group and the ADHD( ) group revealed a poorer performance in general information processing speed for the ADHD(+) group. With regard to set shifting and working memory, no significant differences could be found between the two subgroups. These results suggest that the deficits of the ADHD(total) group are attributable to ADHD rather than to comorbidity. An influence of comorbidity, however, could not be completely ruled out as there was a trend of a poorer performance in the ADHD(+) group on some of the outcome measures. PMID- 21626412 TI - Biological and psychosocial environmental risk factors influence symptom severity and psychiatric comorbidity in children with ADHD. AB - Attention-deficit/hyperactivity disorder (ADHD) is a genetically as well as environmentally determined disorder with a high rate of psychiatric comorbidity. In this study, non-genetic biological and psychosocial risk factors for ADHD symptom severity and comorbid disorders were assessed in 275 children with ADHD, aged 5-13 years, mean age 9.7 (SD 1.9). Pre-/perinatal biological and lifetime psychosocial risk factors as well as data on parental ADHD were obtained. A different pattern of risk factors emerged for inattentive and hyperactive impulsive ADHD symptoms. Inattentive symptoms were strongly influenced by psychosocial risk factors, whereas for hyperactive-impulsive symptoms, predominantly biological risk factors emerged. Hyperactive-impulsive symptoms also were a strong risk factor for comorbid oppositional defiant (ODD) and conduct disorder (CD). Smoking during pregnancy was a risk factor for comorbid CD but not ODD and further differential risk factors were observed for ODD and CD. Comorbid anxiety disorder (AnxD) was not related to ADHD symptoms and additional biological and psychosocial risk factors were observed. This study adds to the body of evidence that non-genetic biological and psychosocial risk factors have an impact on ADHD symptom severity and differentially influence comorbid disorders in ADHD. The findings are relevant to the prevention and treatment of ADHD with or without comorbid disorders. PMID- 21626413 TI - Dietary nitrite and nitrate: a review of potential mechanisms of cardiovascular benefits. AB - PURPOSE: In the last decade, a growing scientific and medical interest has emerged toward cardiovascular effects of dietary nitrite and nitrate; however, many questions concerning their mode of action(s) remain unanswered. In this review, we focus on multiple mechanisms that might account for potential cardiovascular beneficial effects of dietary nitrite and nitrate. RESULTS: Beneficial changes to cardiovascular health from dietary nitrite and nitrate might result from several mechanism(s) including their reduction into nitric oxide, improvement in endothelial function, vascular relaxation, and/or inhibition of the platelet aggregation. From recently obtained evidence, it appears that the longstanding concerns about the toxicity of oral nitrite or nitrate are overstated. CONCLUSION: Dietary nitrite and nitrate may have cardiovascular protective effects in both healthy individuals and also those with cardiovascular disease conditions. A role for nitrite and nitrate in nitric oxide biosynthesis and/or in improving nitric oxide bioavailability may eventually provide a rationale for using dietary nitrite and nitrate supplementation in the treatment and prevention of cardiovascular diseases. PMID- 21626414 TI - Computing an optimal time window of audiovisual integration in focused attention tasks: illustrated by studies on effect of age and prior knowledge. AB - The concept of a "time window of integration" holds that information from different sensory modalities must not be perceived too far apart in time in order to be integrated into a multisensory perceptual event. Empirical estimates of window width differ widely, however, ranging from 40 to 600 ms depending on context and experimental paradigm. Searching for theoretical derivation of window width, Colonius and Diederich (Front Integr Neurosci 2010) developed a decision theoretic framework using a decision rule that is based on the prior probability of a common source, the likelihood of temporal disparities between the unimodal signals, and the payoff for making right or wrong decisions. Here, this framework is extended to the focused attention task where subjects are asked to respond to signals from a target modality only. Evoking the framework of the time-window-of integration (TWIN) model, an explicit expression for optimal window width is obtained. The approach is probed on two published focused attention studies. The first is a saccadic reaction time study assessing the efficiency with which multisensory integration varies as a function of aging. Although the window widths for young and older adults differ by nearly 200 ms, presumably due to their different peripheral processing speeds, neither of them deviates significantly from the optimal values. In the second study, head saccadic reactions times to a perfectly aligned audiovisual stimulus pair had been shown to depend on the prior probability of spatial alignment. Intriguingly, they reflected the magnitude of the time-window widths predicted by our decision theoretic framework, i.e., a larger time window is associated with a higher prior probability. PMID- 21626416 TI - Laser-guidance based cell detection for identifying malignant cancerous cells without any fluorescent markers. AB - Laser guidance technique employs the optical forces generated from a focused Gaussian laser beam incident on a biological cell to trap and guide the cell along the laser propagation direction. The optical force, which determines the guidance speed, is dependent on the cellular characteristics of the cell being guided, such as size, shape, composition and morphology. Different cell populations or subpopulations can be detected without any fluorescent markers by measuring their guidance speeds. We found that cell guidance speeds were sensitive enough to monitor the subtle changes during the progression of mouse fibroblast cells from normal to cancerous phenotype. The results also demonstrated that this technique can effectively distinguish mouse mammary cancerous cells with different metastatic competence. Laser guidance technique can be used as a label-free cell detection method for basic cell biological investigation and cancer diagnosis. PMID- 21626415 TI - Immunosuppressive tumor microenvironment in cervical cancer patients. AB - Cervical cancer is caused by Human papillomavirus (HPV) in virtually all cases. These HPV-induced cancers express the viral oncogenes E6 and E7 and are therefore potentially recognized by the immune system. Despite the abundant presence of these foreign antigens, the immune system is unable to cope with the tumor. Due to the constant immunological pressure, cervical cancers can evolve different immune evasion strategies, which will be described in the current review. Several approaches for immunotherapy of cervical cancer are currently under development, which aim at inducing strong HPV-specific immunity. Besides the reinforcement of potent anti-tumor immune responses, immunotherapy could also enhance HPV-specific T regulatory cells. Supplementary strategies that neutralize an immunosuppressive milieu may have great potential. These strategies are discussed as well. PMID- 21626417 TI - Simple control of fed-batch processes for recombinant protein production with E. coli. AB - A very simple but effective process control technique is proposed that leads to a high batch-to-batch reproducibility with respect to biomass concentration as well as the specific biomass growth rate profiles in E. coli fermentations performed during recombinant protein production. It makes use of the well-established temperature controllers in currently used fermenters, but takes its information from the difference between the controlled culture temperature T (cult) and the temperature T (coolin) of the coolant fed to the fermenter's cooling jacket as adjusted by the fermenter temperature controller. For process control purposes this measured difference is corrected regarding stirrer influences and cumulated before it is used as a new process control variable. As a spin-off of this control, it becomes possible to estimate online the oxygen mass transfer rates and the corresponding k(L)a values during the real cultivation process. PMID- 21626418 TI - Application of Fluolid-Orange-labeled probes for DNA microarray and immunological assays. AB - The usefulness of Fluolid-Orange, a novel fluorescent dye, for DNA microarray and immunological assays has been examined. Fluolid-Orange-labeled probes (DNA and IgG) were stable as examined by laser-photo-bleaching and under heat and dry conditions. Statistical analyses were performed to evaluate the reproducibility of the microarray assay, while stage-specific immunostaining of marker proteins, Kank1 and calretinin, was performed for renal cancers, both giving satisfactory results. The stability of the dye should provide advantages for storing fluorescently labeled probes and re-examining the specimens later in genetic and pathological diagnostics. PMID- 21626419 TI - Macrophages overexpressing VEGF, transdifferentiate into endothelial-like cells in vitro and in vivo. AB - Vascular endothelial growth factor (VEGF) induces endothelial cell differentiation. To investigate the role of VEGF overexpression in regulating the phenotype of macrophages, we transfected mouse macrophages with human VEGF(165) and examined the expression of the genes and proteins for various endothelial markers. Macrophages overexpressing VEGF significantly expressed fetal liver kinase 1 (FLK-1), vascular endothelial cadherin (VE-cadherin), CD31, Von Willebrand factor (vWF), endothelial nitric oxide synthase (eNOS) and CD105. Furthermore, in a model of myocardial infarction (MI), macrophages overexpressing VEGF incorporated into blood vessels. Thus macrophages overexpressing VEGF were transdifferentiate into endothelial-like cells (ELCs) both in vitro and in vivo. PMID- 21626420 TI - [Snake charmer NOTES-proctocolectomy in a male patient]. AB - Restricted ability of triangulation, a highly flexible intestine, as well as limited control and overview over the abdominal organs, complicated extensive abdominal interventions in the past. Thus, additional access sites were needed to allow sufficient interoperative control. The newly developed snake charmer NOTES technique enables the reduction of accesses to the umbilicus and rectum. We report the first case of total proctocolectomy with ileoanal pouch formation in hybrid NOTES technique in a male patient. PMID- 21626421 TI - Sequences and gene organization of the mitochondrial genomes of the liver flukes Opisthorchis viverrini and Clonorchis sinensis (Trematoda). AB - Opisthorchis viverrini and Clonorchis sinensis are important trematodes infecting humans and animals, belonging to the family Opisthorchiidae. In the present study, we sequenced the nearly complete mitochondrial (mt) DNA (mtDNA) sequences of O. viverrini from Laos, obtained the complete mtDNA sequences of C. sinensis from China and Korea, and revealed their gene annotations and genome organizations. The mtDNA sequences of O. viverrini, C. sinensis (China isolate), C. sinensis (Korea isolate) were 13,510, 13,879, and 13,877 bp in size, respectively. Each of the three mt genomes comprises 36 genes, consisting of 12 genes coding for proteins, two genes for rRNA, and 20 genes (O. viverrini) or 22 genes (C. sinensis) for tRNA. The gene content and arrangement are identical to that of Fasciola hepatica, and Paragonimus westermani, but distinct from Schistosoma spp. All genes are transcribed in the same direction and have a nucleotide composition high in T. The contents of A + T of the mt genomes were 59.39% for O. viverrini, 60.03% for C. sinensis (China isolate), and 59.99% for C. sinensis (Korea isolate). Phylogenetic analyses using concatenated amino acid sequences of the 12 protein-coding genes, with three different computational algorithms [maximum parsimony, maximum likelihood, and Bayesian analysis], all revealed distinct groups with high statistical support, indicating that O. viverrini and C. sinensis represent sister taxa. These data provide additional novel mtDNA markers for studying the molecular epidemiology and population genetics of the two liver flukes and should have implications for the molecular diagnosis, prevention, and control of opisthorchiasis and clonorchiasis in humans and animals. PMID- 21626422 TI - Studies on larvicidal and pupicidal activity of Leucas aspera Willd. (Lamiaceae) and bacterial insecticide, Bacillus sphaericus, against malarial vector, Anopheles stephensi Liston. (Diptera: Culicidae). AB - The efficacy of whole plant extracts of Leucas aspera and Bacillus sphaericus has been proven against larvicidal and pupicidal activities of the malarial vector, Anopheles stephensi. The present study investigated the larvicidal and pupicidal activity against the first to fourth instar lavae and pupae of the laboratory reared mosquitoes, A. stephensi. The medicinal plants were collected from the area around Maruthamalai hills, Coimbatore, Tamil Nadu, India. L. aspera whole plant was washed with tap water and shade dried at room temperature. The dried plant materials were powdered by an electric blender. From the powder, 100 g of the plant materials was extracted with 300 ml of organic solvents of ethanol for 8 h using a Soxhlet apparatus. The extracts were filtered through a Buchner funnel with Whatman number 1 filter paper. The crude plant extracts were evaporated to dryness in a rotary vacuum evaporator. The plant extract showed larvicidal and pupicidal effects after 24 h of exposure. All larval instars and pupae have considerably moderate mortality; however, the highest larval mortality was the ethanolic extract of whole plant L. aspera against the first to fourth instar larvae and pupae values of LC(50) = I instar was 9.695%, II instar was 10.272%, III instar was 10.823%, and IV instar was 11.303%, and pupae was 12.732%. B. spaericus against the first to fouth instar larvae and pupae had the following values: I instar was 0.051%, II instar was 0.057%, III instar was 0.062%, IV instar was 0.066%, and for the pupae was 0.073%. No mortality was observed in the control. The present results suggest that the ethanolic extracts of L. aspera and B. sphaericus provided an excellent potential for controlling of malarial vector, A. stephensi. PMID- 21626423 TI - Seroepidemiology of Toxoplasma gondii, Neospora caninum, and Leishmania spp. infections and risk factors for cats from Brazil. AB - The seroprevalence of infection by Toxoplasma gondii, Neospora caninum, and Leishmania spp. was detected through an indirect immunofluorescence in 70 cats from the Andradina Municipality, Sao Paulo State, Brazil. Anti-T. gondii antibodies (titer >64) were detected in 15.7% (11/70) of animals, whereas positivity for N. caninum (titer 16) was not observed in any animal. Of the cats from urban and rural areas, 10.4% (5/48) and 27.2% (6/22) were positive for T. gondii, respectively. Breed, age, food, and contact with animals of other species were significant for considering the positivity for T. gondii (P <= 0.0001). Cats having access to streets (17.1%, 11/64), cats cohabiting with rats (19.6%, 10/51), and cats feeding on homemade food and raw milk (27.2%, 6/22) were positive for T. gondii. In addition, 4.2% (3/70) of the cats were positive for Leishmania spp. by ELISA technique and negative by IFAT without coinfection with T. gondii and Leishmania spp. There was no serological positivity against feline immunodeficiency virus or feline leukemia virus. In conclusion, T. gondii infection in part of the feline population from Andradina is not linked to immunosuppressions or coinfections but probably to postnatal infection in association with the type of diet and presence of rats. PMID- 21626424 TI - Morphology, ultrastructure, genetics, and morphometrics of Diplostomum sp. (Digenea: Diplostomidae) metacercariae infecting the European flounder, Platichthys flesus (L.) (Teleostei: Pleuronectidae), off the northwest coast of Portugal. AB - The morphology, ultrastructure, genetics, and morphometrics of a species of Diplostomum von Nordmann 1832 (Digenea: Diplostomidae), isolated from the European flounder (Platichthys flesus (L.)) caught off the northwest coast of Portugal, are characterized. The metacercarial stage was found unencysted in the lens capsule of the eye. Light microscopical observations revealed the existence of some variability in specimen shape and size, with two morphotypes, referred to as "round" and "long", being apparent. Scanning electron microscopy revealed a smooth, unarmed tegument, with the lappet region being the most irregular and porose. Both the oral and ventral suckers were provided with a series of papillae, which presented very distinctive ultrastructural features and were particularly conspicuous in the case of the ventral sucker. The two morphotypes detected were found to have 100% genetic correspondence in the 18S + ITS1 + 5.8S region of the rDNA. Since the genetic data for this metacercaria differed from those of the species of Diplostomum available in GenBank, a description of a new genotype (accession number GQ370809) is provided. The molecular phylogenetic analyses, in conjunction with principal components and cluster analyses based on morphometric data, revealed the existence of consistent differences between the Diplostomum sp. metacercariae from flounder compared with Diplostomum spathaceum, Diplostomum mergi, Diplostomum pseudospathaceum, and Diplostomum paracaudum. The latter of these species was found to be the most similar to the present material. Our results do not support an evolutionary separation of the European and North American species of Diplostomum. PMID- 21626425 TI - Mosquito species abundance and diversity in Malindi, Kenya and their potential implication in pathogen transmission. AB - Mosquitoes (Diptera: Culicidae) are important vectors of human disease-causing pathogens. Mosquitoes are found both in rural and urban areas. Deteriorating infrastructure, poor access to health, water and sanitation services, increasing population density, and widespread poverty contribute to conditions that modify the environment, which directly influences the risk of disease within the urban and peri-urban ecosystem. The objective of this study was to evaluate the mosquito vector abundance and diversity in urban, peri-urban, and rural strata in Malindi along the Kenya coast. The study was conducted in the coastal district of Malindi between January and December 2005. Three strata were selected which were described as urban, peri-urban, and rural. Sampling was done during the wet and dry seasons. Sampling in the wet season was done in the months of April and June to cover the long rainy season and in November and December to cover the short rainy season, while the dry season was between January and March and September and October. Adult mosquito collection was done using Pyrethrum Spray Collection (PSC) and Centers for Disease Control and Prevention (CDC) light traps inside houses and specimens were identified morphologically. In the three strata (urban, peri-urban, and rural), 78.5% of the total mosquito (n = 7,775) were collected using PSC while 18.1% (n = 1,795) were collected using the CDC light traps. Using oviposition traps, mosquito eggs were collected and reared in the insectary which yielded 329 adults of which 83.8% (n = 276) were Aedes aegypti and 16.2% (n = 53) were Culex quinquefasciatus. The mosquito distribution in the three sites varied significantly in each collection site. Anopheles gambiae, Anopheles funestus and Anopheles coustani were predominant in the rural stratum while C. quinquefasciatus was mostly found in urban and peri-urban strata. However, using PSC and CDC light trap collection techniques, A. aegypti was only found in urban strata. In the three strata, mosquitoes were mainly found in high numbers during the wet season. Further, A. gambiae, C. quinquefasciatus, and A. aegypti mosquitoes were found occurring together inside the houses. This in turn exposes the inhabitants to an array of mosquito-borne diseases including malaria, bancroftian filariasis, and arboviruses (dengue fever, Yellow fever, Rift Valley fever, Chikungunya fever, and West Nile Virus). In conclusion, our findings provide useful information for the design of integrated mosquito and disease control programs in East African environments. PMID- 21626426 TI - Frailty and multiple comorbidities in the elderly patient with heart failure: implications for management. AB - Heart failure (HF) in the elderly is a major public health problem, and its prevalence is rising. Outcomes of HF in the elderly have not changed in the past 2 decades despite the introduction of novel HF therapies. This may be due to the combined impact of multiple comorbidities and frailty. The majority of elderly patients with HF are frail with multiple comorbidities. These comorbidities, along with frailty, contribute to the poor outcome of HF in the elderly and pose independent management challenges. More research is needed to better understand the interaction between frailty and multiple comorbidities and the mechanisms by which they impact HF and its management; develop prognostic tools that incorporate frailty and multiple comorbidities and provide more accurate prediction of outcomes; test available treatments in typical elderly patients; and develop and test novel interventions that directly address the adverse impact of multiple comorbidities and frailty. PMID- 21626427 TI - Plication of the right atrium in order to confront a right coronary artery under tension graft. AB - An under tension graft to the right coronary artery may result in graft spasm, hypoperfusion and myocardial infarction. We suggest plication of the right atrium in order to confront the under tension graft (either arterial or venous), if tension is due to a shorter (up to 2 cm) graft than one needed. We present this technique applied to 10 of our patients. The radial artery was the under tension graft in two cases and a saphenous vein in eight. Perioperative and postoperative arrhythmias, myocardial infarction, hemorrhage and jugular or hepatic congestion were not observed. Central vein pressure remained normal. Tricuspid valve- estimated by postoperative echo--was undisturbed. Plication of the right atrium in case of an under tension graft is a quick, simple and safe technique. PMID- 21626428 TI - Comparison of two models for predicting non-sentinel lymph node metastases in sentinel lymph node-positive breast cancer patients. AB - Several models for the prediction of non-sentinel lymph node (SLN) metastasis in SLN-positive breast cancer patients have been proposed. In this study, the authors evaluate the Memorial Sloan-Kettering Cancer Center (MSKCC) nomogram and an axilla scoring system from Tenon Hospital to predict the probability of having non-(SLN) involvement and to define their actual usefulness when applied to the subgroup of patients with micrometastasis in SLN. The study population consisted of 103 patients: 74 with macrometastasis and 29 with micrometastasis in the SLN. The receiver operating characteristic (ROC) curves were drawn and the area under the curves (AUCs) was calculated to assess the discriminative power of the nomograms. Both the ROCs and relative AUCs were calculated for all the patients in the study and for the two subgroups. The AUC for the entire study population was 0.712 in the MSKCC nomogram and 0.759 in the Tenon score. In 74 patients with macrometastasis in SLN, the AUC was 0.760 in MSKCC nomogram and 0.707 in Tenon score. Micrometastasis in SLN was found in t29 patients: AUC was 0.577 in the MSKCC nomogram and 0.738 in the Tenon score. Several institutions have tested the MSKCC nomogram, with AUC ranging from 0.58 to 0.84. It was not validated by four studies, which did not recommend its use in patients with micrometastasis. In our results, the validation of the Tenon score confirmed its relevance even in patients with micrometastasis in SLN. PMID- 21626429 TI - Massive upper gastrointestinal hemorrhage caused by an arterio-esophageal fistula. PMID- 21626430 TI - Acquired hemophilia: a rare but life-threatening potential cause of bleeding in the intensive care unit. AB - OBJECTIVE: There are a number of potential etiologies of severe bleeding encountered in the intensive care unit. Although rare, acquired hemophilia is one such etiology that often presents with major bleeding requiring intensive care. Despite the introduction of effective treatments, the reported mortality rate of patients with acquired hemophilia ranges from 6 to 8% and is in part attributable to sequential delays in diagnosis and appropriate treatment. The purpose of this review is to familiarize the intensive care specialist with this underrecognized cause of bleeding, with an emphasis on diagnosis and treatment. METHODS: As the objective of this article was to provide a concise overview of the diagnosis and management of acquired hemophilia, a directed search of English-language literature was undertaken using the PubMed database, targeting such topics as the differential diagnosis of bleeding in the intensive care unit and the epidemiology, diagnosis, and treatment of acquired hemophilia. Clinical study findings pertaining to the efficacy of specific treatments for acquired hemophilia were summarized. RESULTS AND CONCLUSION: Recognition of acquired hemophilia presents a clinical challenge, given the rarity of this condition, a general lack of familiarity with acquired hemophilia, and the potential for confusion with other more common causes of bleeding in the intensive care unit. Nevertheless, there are sentinel clinical and laboratory findings that should raise suspicion of this diagnosis. The treatment of acquired hemophilia is a multi-step, physiologically focused process aimed at controlling both active and recurrent bleeding. Therefore, prompt diagnosis is central to prognosis. Consultation with a hematologist may facilitate efficient diagnosis and management. PMID- 21626431 TI - Levosimendan attenuates pulmonary vascular remodeling. AB - PURPOSE: The calcium-sensitizing drug levosimendan increases myocardial contractility and, by activating K(+)-channels, dilates pulmonary vessels. In the acute setting, levosimendan is clinically used to treat right heart failure in pulmonary hypertension. As K(+)-channel activation elicits several beneficial effects in the vascular system, we hypothesized that levosimendan also attenuates the remodeling process in the monocrotaline model of rat pulmonary hypertension. METHODS AND RESULTS: Animal subgroups received levosimendan, the K(+)-channel opener nicorandil, or levosimendan together with the K(+)-adenosine triphosphate (ATP)-sensitive potassium channel (K(ATP)) blocker glibenclamide. Morphometric analyses revealed that levosimendan and nicorandil attenuated the increased pulmonary vascular medial wall thickness after monocrotaline challenge. Accordingly, in vivo BrdU assays revealed that levosimendan significantly diminished proliferation of pulmonary arterial smooth muscle cells (PASMCs), and this effect was attenuated by glibenclamide. Levosimendan also reduced right ventricular hypertrophy, but this effect was not glibenclamide sensitive and not recapitulated by nicorandil. In cell culture, levosimendan had a direct inhibitory effect on the platelet-derived growth factor (PDGF)-induced proliferation of PASMCs, which however required high concentrations of the compound, pointing towards an endothelial effect. Indeed, levosimendan increased cyclic guanosine monophosphate (cGMP) in human umbilical vein endothelial cells (HUVECs) and impaired the tumor necrosis factor-alpha (TNF-alpha)-induced inflammatory expression of E-selectin, intercellular adhesion molecule-1 (ICAM 1), cyclooxygenase-2 (COX-2), and monocyte chemotactic protein-1 (MCP-1). In luciferase reporter gene assays in HUVECs, levosimendan dose-dependently attenuated the TNF-alpha-stimulated increase of proinflammatory transcription factors activator protein 1 (AP1), hypoxia-inducible factor-1alpha (HIF-1alpha), and nuclear factor-kappaB (NF-kappaB). CONCLUSIONS: Levosimendan attenuates pulmonary vascular remodeling, presumably by an antiproliferative and anti inflammatory effect which is mediated by cellular hyperpolarization. The compound also has a direct inhibitory effect on cardiac hypertrophy, which is however K(+) channel independent. PMID- 21626432 TI - Continuous arterial pressure waveform monitoring in pediatric cardiac transplant, cardiomyopathy and pulmonary hypertension patients. AB - PURPOSE: A continuous cardiac output monitor based on arterial pressure waveform (FloTrac/Vigileo; Edwards Lifesciences, Irvine, CA) is now approved for use in adults but not in children. This device is minimally invasive, calculates cardiac output continuously and in real time, and is easy to use. Our study sought to validate the FloTrac with the pulmonary artery catheter (PAC) intermittent thermodilution technique in pediatric cardiac patients. METHODS: This was a prospective pilot study comparing cardiac output measurements obtained via the FloTrac and arterial pressure waveform analysis with intermittent thermodilution. Subjects carried the diagnosis of pulmonary hypertension or cardiomyopathy, or were in the postoperative course after orthotopic heart transplantation. RESULTS: Enrolled in the study were 31 subjects, and 136 data points were obtained. The age range was 8 months to 16 years. The mean body surface area (BSA) was 1.1 m(2). Bland-Altman plots for the mean cardiac outputs of all subjects with a BSA >= 1 m(2) showed limits of agreement of -2.7 to 8.0 l/min (+/- 5.4 l/min). Patients with a BSA <= 1 m(2) demonstrated even wider limits of agreement (+/- 8.5 l/min). The intraclass correlation for the PAC was 0.929 and 0.992 for the FloTrac. CONCLUSION: There was poor agreement between the PAC and FloTrac in measuring cardiac output in a population of children with pulmonary hypertension or cardiomyopathy, or after cardiac transplantation. This is in contrast to adult studies published thus far. This suggests that the utility of the FloTrac and measurements obtained from arterial pulse wave analysis in children is uncertain at this time. PMID- 21626433 TI - The Elliott Youth Development lecture. PMID- 21626434 TI - Invited commentary: childhood and adolescent obesity: psychological and behavioral issues in weight loss treatment. AB - The prevalence of childhood and adolescent obesity has tripled in the past three decades. This increase has been accompanied by a dramatic rise in obesity-related health complications among American youth. Thus, many obese youth are now experiencing illnesses that will threaten their life expectancy in the absence of significant weight loss. Despite these concerns, a relatively modest body of research has focused on the treatment of adolescent obesity. Results from trials investigating the efficacy of behavioral and pharmacological treatments, like studies of these interventions with adults, suggest that individuals typically lose 5-10% of their initial weight. Unfortunately, weight regain is common. Given the increase in the number of obese adolescents, coupled with the modest results from more conservative treatment approaches, it is not surprising that bariatric surgery for adolescents who suffer from extreme obesity has grown in popularity. The weight losses after surgery are impressive and many adolescents, like adults, experience significant improvements in their physical and mental health postoperatively. However, only a small fraction of adolescents and adults who are heavy enough for bariatric surgery present for surgical treatment. Among those who undergo surgery, a significant minority appear to struggle with a number of behavioral and psychosocial issues that threaten their lifelong success. With all of this in mind, the current obesity problem in the United States and other Westernized countries likely will present a significant challenge to both current and future medical and mental health professionals who work with adolescents and young adults. PMID- 21626435 TI - The molecular mechanism underlying morphine-induced Akt activation: roles of protein phosphatases and reactive oxygen species. AB - Although Akt is reported to play a role in morphine's cardioprotection, little is known about the mechanism underlying morphine-induced Akt activation. This study aimed to define the molecular mechanism underlying morphine-induced Akt activation and to determine if the mechanism contributes to the protective effect of morphine on ischemia/reperfusion injury. In cardiac H9c2 cells, morphine increased Akt phosphorylation at Ser(473), indicating that morphine upregulates Akt activity. Phosphatase and tensin homolog deleted on chromosome 10 (PTEN), a major regulator of the phosphatidylinositol 3-kinase (PI3K)/Akt signaling, was not involved in the action of morphine on Akt activity. Morphine decreased the activity of PP2A, a major protein Ser/Thr phosphatase, and inhibition of PP2A with okadaic acid (OA) mimicked the effect of morphine on Akt activity. The effects of morphine on PP2A and Akt activities were inhibited by the reactive oxygen species (ROS) scavenger N-(2-mercaptopropionyl)glycine (MPG) and the mitochondrial K(ATP) channel closer 5-hydroxydecanoate (5HD). In support, morphine could produce ROS and this was reversed by 5HD. Finally, the cardioprotective effect of morphine on ischemia-reperfusion injury was mimicked by OA but was suppressed by 5HD or MPG, indicating that protein phosphatases and ROS are involved in morphine's protection. In conclusion, morphine upregulates Akt activity by inactivating protein Ser/Thr phosphatases via ROS, which may contribute to the cardioprotective effect of morphine. PMID- 21626436 TI - Polymorphism of M307 of the FUT1 gene and its relationship with some immune indexes in Sutai pigs (Duroc x Meishan). AB - The alpha (1,2)fucosyltransferase (FUT1) gene has been identified as a candidate gene for controlling the expression of the enterotoxigenic Escherichia coli (ETEC) F18 receptor. Polymorphisms were detected at the M307 position in FUT1 of a breeding base population of Sutai pigs and their correlations to immune parameters analyzed. After digestion by Hin6I, three genotypes were identified at M307, AA (frequency 0.235), AG (0.609), and GG (0.156), with significant deviation from Hardy-Weinberg equilibrium (P < 0.01). The hemoglobin and white blood cell count of the AA genotype pigs were significantly higher than those of AG and GG pigs (P < 0.05). The results indicated that AA pigs not only are resistant to edema disease and post-weaning diarrhea in piglets but also have relatively strong resistance to disease in general. PMID- 21626437 TI - Impact of low percentage of data below the quantification limit on parameter estimates of pharmacokinetic models. AB - The objectives of the simulation study were to evaluate the impact of BQL data on pharmacokinetic (PK) parameter estimates when the incidence of BQL data is low (e.g. <=10%), and to compare the performance of commonly used modeling methods for handling BQL data such as data exclusion (M1) and likelihood-based method (M3). Simulations were performed by adapting the method of a recent publication by Ahn et al. (J Phamacokinet Pharmacodyn 35(4):401-421, 2008). The BQL data in the terminal elimination phase were created at frequencies of 1, 2.5, 5, 7.5, and 10% based on a one- and a two-compartment model. The impact of BQL data on model parameter estimates was evaluated based on bias and imprecision. The simulations demonstrated that for the one-compartment model, the impact of ignoring the low percentages of BQL data (<=10%) in the elimination phase was minimal. For the two compartment model, when the BQL incidence was less than 5%, omission of the BQL data generally did not inflate the bias in the fixed-effect parameters, whereas more pronounced bias in the estimates of inter-individual variability (IIV) was observed. The BQL data in the elimination phase had the greatest impact on the volume of distribution estimate of the peripheral compartment of the two compartment model. The M3 method generally provided better parameter estimates for both PK models than the M1 method. However, the advantages of the M3 over the M1 method varied depending on different BQL censoring levels, PK models and parameters. As the BQL percentages decreased, the relative gain of the M3 method based on more complex likelihood approaches diminished when compared to the M1 method. Therefore, it is important to balance the trade-off between model complexity and relative gain in model improvement when the incidence of BQL data is low. Understanding the model structure and the distribution of BQL data (percentage and location of BQL data) allows selection of an appropriate and effective modeling approach for handling low percentages of BQL data. PMID- 21626438 TI - Molecular imaging with 68Ga-SSTR PET/CT and correlation to immunohistochemistry of somatostatin receptors in neuroendocrine tumours. AB - PURPOSE: Somatostatin receptors (SSTR) are known for an overexpression in gastroenteropancreatic neuroendocrine tumours (GEP-NET). The aim of the present study was to find out if the receptor density predicted by the semi-quantitative parameters generated from the static positron emission tomography (PET/CT) correlated with the in vitro immunohistochemistry using a novel rabbit monoclonal anti-SSTR2A antibody (clone UMB-1) for specific SSTR2A immunohistochemistry and polyclonal antibodies for SSTR1 and 3-5. METHODS: Overall 14 surgical specimens generated from 34 histologically documented GEP-NET patients were correlated with the preoperative (68)Ga-DOTA-NOC PET/CT. Quantitative assessment of the receptor density was done using the immunoreactive score (IRS) of Remmele and Stegner; the additional 4-point IRS classification for immunohistochemistry and standardized uptake values (SUV(max) and SUV(mean)) were used for PET/CT. RESULTS: The IRS for SSTR2A and SSTR5 correlated highly significant with the SUV(max) on the PET/CT (p < 0.001; p < 0.05) and the IRS for SSTR2A with the SUV(mean) (p < 0.013). The level of SSTR2A score correlated significantly with chromogranin A staining and indirectly to the tumour grading. CONCLUSION: The highly significant correlation between SSTR2A and SSTR5 and the SUV(max) on the (68)Ga-DOTA-NOC PET/CT scans is concordant with the affinity profile of (68)Ga-DOTA-NOC to the SSTR subtypes and demonstrates the excellent qualification of somatostatin analogues in the diagnostics of NET. This study correlating somatostatin receptor imaging using (68)Ga-DOTA-NOC PET/CT with immunohistochemically analysed SSTR also underlines the approval of therapy using somatostatin analogues, follow-up imaging as well as radionuclide therapy. PMID- 21626439 TI - Distrust in the healthcare system and organ donation intentions among African Americans. AB - The purpose of this study is to further understanding of the association between distrust in the healthcare system and written and verbal expressions of donation intentions among African Americans. We hypothesize that distrust in the healthcare system will be significantly, positively associated with both verbal and written donation intentions. Five hundred and eighty five participants completed a 98-item survey that included scales on distrust in the healthcare system and donation intentions. Bivariate analyses (t-tests, ANOVA, chi-square tests and odds ratios) were used to explore the extent to which donation intentions and distrust in the healthcare system varied by demographic characteristics and the association between the distrust in the healthcare system scale and verbal and written donation intentions. Separate logistic regressions were performed with each of the dependent variables to see if significant associations remained while controlling for confounders. Findings based on the multiple regression indicate that when controlling the participant's education level, distrust in the healthcare system was not significantly related to written donation intentions (OR = 1.04; P = .12). When controlling for education level, health insurance status, Community Health Advocates group and marital status, distrust in the healthcare system was significantly associated with verbal donation intentions (OR = 1.08; P < 0.05). Our results suggest that distrust in the healthcare system varies in the way that it is associated with donation intentions. Future organ donation studies should be conducted to determine the pathways through which distrust in the healthcare system impacts different types of organ donation intentions. PMID- 21626440 TI - Chinese origin rhesus macaque major histocompatibility complex class I molecules promiscuously present epitopes from SIV associated with molecules of Indian origin; implications for immunodominance and viral escape. AB - The presentation of identical peptides by different major histocompatibility complex class I (MHC-I) molecules, termed promiscuity, is a controversial feature of T cell-mediated immunity to pathogens. The astounding diversity of MHC-I molecules in human populations, presumably to enable binding of equally diverse peptides, implies promiscuity would be a rare phenomenon. However, if it occurs, it would have important implications for immunity. We screened 77 animals for responses to peptides known to bind MHC-I molecules that were not expressed by these animals. Some cases of supposed promiscuity were determined to be the result of either non-identical optimal peptides or were simply not mapped to the correct MHC-I molecule in previous studies. Cases of promiscuity, however, were associated with alterations of immunodominance hierarchies, either in terms of the repertoire of peptides presented by the different MHC-I molecules or in the magnitude of the responses directed against the epitopes themselves. Specifically, we found that the Mamu-B*017:01-restricted peptides Vif HW8 and cRW9 were also presented by Mamu-A2*05:26 and targeted by an animal expressing that allele. We also found that the normally subdominant Mamu-A1*001:01 presented peptide Gag QI9 was also presented by Mamu-B*056:01. Both A2*05:26 and B*056:01 are molecules typically or exclusively expressed by animals of Chinese origin. These data clearly demonstrate that MHC-I epitope promiscuity, though rare, might have important implications for immunodominance and for the transmission of escape mutations, depending on the relative frequencies of the given alleles in a population. PMID- 21626441 TI - Primary esophageal small cell carcinoma with brain metastasis and with CD56, KIT, and PDGFRA expressions. PMID- 21626442 TI - Downregulated expression of inhibitor of growth 4 (ING4) in advanced colorectal cancers: a non-randomized experimental study. AB - Colorectal cancer has a high cure rate if it can be detected early. Identifying and understanding the genes involved may enable early diagnosis and reduce mortality. The aim of our study was to investigate the correlation between the expression of ING4 and the pathological features in patients with colorectal cancer. We assayed ING4 protein expression levels in tumor samples from 97 patients diagnosed with colorectal cancer between January 2001 and January 2002. The patients received no other treatments except surgery. ING4 protein expression was downregulated in adenoma relative to normal mucosa and further reduced in colorectal cancer tissues. Furthermore, the suppression of ING4 expression was also related to the more advanced Dukes' stages. We observed that ING4 expression levels in patients with lymphatic metastasis were lower than those without metastasis. Together, our results indicate that ING4 play a role in colorectal carcinoma progression. PMID- 21626443 TI - Comparison of initial hydrolysis of the three dimethyl phthalate esters (DMPEs) by a basidiomycetous yeast, Trichosporon DMI-5-1, from coastal sediment. AB - PURPOSE: Dimethyl phthalate esters (DMPEs) are a group of plasticizers commonly detected in the environment with potential adverse human health impact. The degradation of DMPEs by fungal systems has been studied to a limited extent, particularly by yeasts. In this study, a basidiomycetous yeast Trichosporon DMI-5 1 capable of degrading DMPEs was obtained and the degradation pathways were investigated. RESULTS: A DMPE-degrading yeast was isolated from costal sediment by enrichment culture technique and was identified as Trichosporon sp. DMI-5-1 based on microscopic morphology and 18S rDNA sequence. Comparative investigations on biodegradation of three isomers of DMPEs, namely dimethyl phthalate (DMP), dimethyl isophthalate (DMI), and dimethyl terephthalate (DMT), were carried out with this yeast strain. Trichosporon sp. DMI-5-1 could not mineralize DMPEs completely but transform them to respective monomethyl phthalate or phthalic acid. Biochemical degradation pathways for the three DMPE isomers by Trichosporon sp. DMI-5-1 were apparently different. The yeast carried out one-step ester hydrolysis of DMP and DMI to respective monoesters (monomethyl phthalate and monomethyl isophthalate, respectively) and no further metabolism of these two monoesters. Meanwhile, DMT was transformed by the yeast to monomethyl terephthalate and subsequently to terephthalic acid by stepwise hydrolysis of the two ester bonds. CONCLUSIONS: This study shows that different catalytic processes are involved in the transformation of DMPEs by the basidiomycetous yeast Trichosporon sp. DMI-5-1 and suggests that its esterases, responsible for the initial hydrolyzing the two ester bonds of DMPEs, are highly substrate specific. PMID- 21626444 TI - Training addiction counselors to implement CBT for depression. AB - Many clients in publicly funded substance abuse treatment programs suffer from depression yet lack access to effective mental health treatment. This study sought to examine whether addiction counselors could be effectively trained to deliver group CBT for depression and to ascertain client perceptions of the treatment. Five counselors were trained in the therapy and treated 113 clients with depression symptoms. Counselors demonstrated high fidelity to the therapy and client perceptions of the therapy were positive. Our results suggest that training addiction counselors to deliver group CBT for depression is a promising integrated treatment approach for co-occurring depression and substance disorders. PMID- 21626445 TI - Causes of headache in patients with a primary diagnosis of sinus headache. AB - Headache is a common occurrence among the general population. Although the pain could be a symptom of acute sinusitis, chronic sinusitis is not considered as a usual cause of headache. In addition, autonomic-related symptoms in the sinonasal region may be associated with vascular pain. Confusion regarding these symptoms could lead to an incorrect diagnosis of sinusitis. A prospective cross-sectional study was conducted at two tertiary referral centers with residency programs in otorhinolaryngology, head and neck surgery and neurology. The study included 58 patients with a diagnosis of "sinus headache" made by a primary care physician. Exclusion criteria were as follows: previous diagnosis of migraine or tension type headache; evidence of sinus infection during the past 6 months; and the presence of mucopurulent secretions. After comprehensive otorhinolaryngologic and neurologic evaluation, appropriate treatment was started according to the final diagnosis and the patient was assessed monthly for 6 months. The final diagnoses were migraine, tension-type headache and chronic sinusitis with recurrent acute episodes in 68, 27 and 5% of the patients, respectively. Recurrent antibiotic therapy was received by 73% of patients with tension-type headache and 66% with migraine. Sinus endoscopy was performed in 26% of the patients. Therapeutic nasal septoplasty was performed in 16% of the patients with a final diagnosis of migraine, and 13% with tension-type headache. Many patients with self-described or primary care physician labeled "sinus headache" have no sinonasal abnormalities. Instead, most of them meet the IHS criteria for migraine or tension-type headache. PMID- 21626446 TI - Tensor fascia lata flap reconstruction following groin dissection: is it worthwhile? AB - OBJECTIVE: To compare the morbidity of primary skin closure with elective Tensor Fascia Lata (TFL) flap cover in groin dissections. MATERIALS AND METHODS: This was a retrospective study between January 2007 and December 2009. All patients undergoing groin dissections without skin involvement were included. RESULTS: Of the twenty-five patients, who underwent groin dissections, 14 had primary skin closure (28 groin dissections)-group I. Eleven had TFL flap cover as a means of primary reconstruction (20 groin dissections)-group II. In group I, there were 16 (57%) inguinal dissections and 12 (43%) ilioinguinal block dissections, whereas 82% in group II underwent ilioinguinal dissections (p = 0.09). Wound infection requiring treatment with a culture specific antibiotic was required in 4 (14%) in group I (n = 28) and only 1 (5%) in group II (n = 20) (p = 0.38). In group I, 7 (25%) had major flap necrosis and minor necrosis was seen in another 7 (25%). Only three (15%) in group II developed minor flap necrosis (p = 0.01). Following an ilioinguinal dissection, flap necrosis occurred in 75% of groins that underwent primary closure and in 17% of those which were reconstructed with TFL (p = 0.001). Seroma formation was seen in 5 (18%) in group I and 3 (15%) in group II (p = 1.0). Lymphoedema occurred in equal numbers in both groups. The duration of hospital stay was 20 +/- 14 days in the primary closure group and 16 +/- 3 days in the TFL group. CONCLUSION: The TFL flap can reduce postoperative morbidity and decrease hospital stay. Prophylactic TFL flap reconstruction following ilioinguinal dissections is advisable. PMID- 21626447 TI - Changes in osteocyte density correspond with changes in osteoblast and osteoclast activity in an osteoporotic sheep model. AB - Histomorphometric assessment of trabecular bone in osteoporotic sheep showed that bone volume, osteoid surface area, bone formation rate, and osteocyte density were reduced. In contrast, eroded surface area and empty lacunae density were increased. Changes in osteocyte density correlated with changes in osteoblast and osteoclast activity. INTRODUCTION: Osteocytes contribute to the regulation of the activity of osteoclasts and osteoblasts that together control bone mass. Osteocytes therefore likely play a role in the loss of bone mass associated with osteoporosis. The purpose of this study was to investigate the relationships between osteocyte lacunar density and other bone histomorphometric parameters in the iliac crest (IC) and lumbar spine (LS) of osteoporotic sheep. METHODS: Osteoporosis was induced in ten mature ewes by an established protocol involving a combination of ovariectomy, dexamethasone injection, and low calcium diet for 6 months. Five ewes were used as controls. Post-mortem IC and LS biopsies were collected and processed for further histomorphometric assessment. RESULTS: Bone volume, osteoid surface, and bone formation rate in the IC and LS of osteoporotic sheep were reduced compared to those of the controls. In contrast, eroded surface area was increased in osteoporotic sheep. In the osteoporotic group, osteocyte density was reduced in the LS region and to a greater extent in the IC region. The empty osteocyte lacunae were increased 1.7-fold in LS and 2.1-fold in IC in the osteoporotic group. The osteocyte density correlated positively with markers of osteoblast activity and negatively with those of osteoclast activity. CONCLUSIONS: Depletion of osteocytes and an increase in the empty lacunae could be important factors contributing to bone loss in this model since they may adversely affect intercellular communication between osteoblasts and osteoclasts. The regional differences in histology suggest that there may be different pathological mechanisms operating at different anatomical sites. PMID- 21626448 TI - The state of cancer epidemiology curricula in postgraduate schools worldwide. AB - The study aimed to describe the cancer epidemiology curricula in postgraduate schools worldwide. Using a stepwise approach, information on the cancer epidemiology curricula were abstracted through an internet search of medical or public heath schools worldwide. The common scientific outline (a scholarly developed classification of cancer-related topics) was used to describe the extents that cancer epidemiology and its scientific domains are incorporated into postgraduate degrees in the epidemiology. Among the 120 studied schools, no school offered an explicitly doctoral degree in cancer epidemiology. Just eight schools offered cancer epidemiology as an area of concentration in their epidemiology curricula. The contents of the cancer epidemiology courses offered in different schools were related in 44% of times to topics of cancer control, 19% times to risk factors, and just 11% of times to biology of cancer. The need for comprehensive re-evaluation of the cancer epidemiology curricula in postgraduate teaching was concluded. PMID- 21626449 TI - Effects of a 1-week inpatient course including information, physical activity, and group sessions for prostate cancer patients. AB - This study aims to explore the effects of a 1-week inpatient course including information, physical activity (PA), and group sessions on physical and mental health-related outcomes for prostate cancer (PCa) patients. Further to assess the patients' satisfaction with the course. PCa patients completed a questionnaire assessing PA, fatigue, mental distress, and quality of life 1 month before (T0) and 3 months after (T1) the course. Total fatigue, physical fatigue, and PSA anxiety decreased significantly from T0 to T1. No significant changes were observed in the other measures. The majority of the participants were satisfied with the course. In spite of minor reductions in fatigue and PSA anxiety and satisfied patients, the findings indicate that a 1-week inpatient course does not influence substantially on most of the health-related outcomes in PCa patients 3 months after the course. PMID- 21626450 TI - Detection and characterization of a ST6 clone of vanB2-Enterococcus faecalis from three different hospitals in Spain. AB - Thirteen vancomycin-resistant and teicoplanin-susceptible Enterococcus faecalis isolates were recovered from unrelated patients in three Spanish hospitals from November 2009 to December 2010. All isolates carried the vanB2 gene, showed indistinguishable or closely-related PFGE patterns and were ascribed to the sequence type ST6 (included into the high-risk clonal-complex CC2). They showed a multiresistance phenotype (erythromycin, tetracycline, ciprofloxacin and high level-resistance to streptomycin, gentamicin and kanamycin) and harboured the aac(6')-aph(2"), ant(6)-Ia, and tet(M)+/-tet(L) genes. All isolates produced gelatinase and harboured the gelE gene, but not the esp or hyl genes. The inclusion of the vanB2 gene into the Tn5382 transposon was demonstrated in one isolate. Clonal dissemination of vanB2-containing the E. faecalis strain is demonstrated. PMID- 21626452 TI - Validation of the Japanese version of the quality of recovery score QoR-40. AB - PURPOSE: The quality of recovery score QoR-40 is a recovery-specific and patient rated questionnaire to assess the early postoperative health status of patients. However, the Japanese version of the QoR40 has not been established. The aim of this study was to validate the quality of recovery 40 Japanese version (the QoR 40J) according to the methods adopted by the International Quality of Life Assessment (IQOLA) project. METHODS: After obtaining ethics committee approval and consent, 192 general and otological surgical patients were studied. The QoR 40J was used to measure postoperative health status on day 3 and 1 month after surgery. The level of quality of life was also evaluated using a general, health related quality of life questionnaire (Short-Form Health Survey-36 sub-scales; SF 36), at 1 month after the surgery. Psychometric analysis including the following properties: test-retest reliability, internal consistency, predictive validity, and measurement of responsiveness, was performed to validate the QoR-40J. RESULTS: Test-retest reliability (Spearman's correlation coefficient) and internal consistency (Cronbach's alpha) of the QoR-40J were 0.887 and 0.91, respectively. A significant relationship was observed between the total QoR-40J score and duration of hospitalization (r = -0.291) and between the global QoR-40J score and postoperative scores of the SF-36 sub-scales (physical function, rho = 0.287; vital score, rho = 0.349). The standardized mean of the QoR-40J, a measurement of responsiveness, was 0.70. CONCLUSION: The results of the psychometric analysis indicated that the QoR-40J has characteristics of acceptable validity, reliability, and responsiveness in clinical practice in Japan. The QoR-40J may aid in evaluating the quality of recovery after surgery or the quality of methods of anesthesia. PMID- 21626451 TI - The diabetic pregnancy and offspring BMI in childhood: a systematic review and meta-analysis. AB - AIMS/HYPOTHESIS: Offspring of mothers with diabetes are at increased risk of metabolic disorders in later life. Increased offspring BMI is a plausible mediator. We performed a systematic review and meta-analysis of studies examining offspring BMI z score in childhood in relation to maternal diabetes. METHODS: Papers reporting BMI z scores for offspring of diabetic (all types, and pre- and during-pregnancy onset) and non-diabetic mothers were included. Citations were identified in PubMed; bibliographies of relevant articles were hand-searched and authors contacted for additional data where necessary. We compared offspring BMI z score with and without adjustment for maternal pre-pregnancy BMI. We performed fixed effect meta-analysis except where significant heterogeneity called for use of a random effects analysis. RESULTS: Data were available from nine studies. In the diabetic group unadjusted mean offspring BMI z score was 0.28 higher (all diabetic mothers vs controls (95% CI 0.09, 0.47; p = 0.004; nine studies; offspring of diabetic mothers n = 927, controls n = 26,384) and with adjustment for maternal pre-pregnancy BMI, 0.07 higher (95% CI -0.15, 0.28; p = 0.54; three studies; offspring of diabetic mothers n = 244, controls n = 11,206). There was no evidence of a difference in offspring BMI z score in relation to type of diabetes (gestational vs type 1, p = 0.95). CONCLUSIONS/INTERPRETATION: Maternal diabetes is associated with increased offspring BMI z score, although this is no longer apparent after adjustment for maternal pre-pregnancy BMI in the limited number of studies in which this is reported. Causal mediators of the effect of maternal diabetes on offspring outcomes remain to be established; we recommend that future research includes adjustment for maternal pre-pregnancy BMI. PMID- 21626453 TI - Change in left ventricular geometry during antihypertensive treatment in children with primary hypertension. AB - The pattern of the left ventricle (LV) has important significance in adults with hypertension. The aim of the present study was to analyze changes and determinants of LV geometry after 1 year of antihypertensive treatment in children with primary hypertension (PH) in relation to metabolic abnormalities and anthropometrical parameters. In 86 children (14.1 +/- 2.4 years) with newly diagnosed PH, LV geometry and biochemical parameters before and after 12 months of standard antihypertensive therapy were assessed. At baseline, normal LV geometry (NG) was found in 42 (48.9%), concentric remodeling (CR) in 4 (4.6%), concentric hypertrophy (CH) in 8 (9.3%), and eccentric hypertrophy (EH) in 32 (37.2%) patients. The prevalence of NG in patients with severe hypertension was significantly lower than in patients with ambulatory hypertension. There were no differences in dipping status in relation to LV geometry. Patients with CH and EH were more viscerally obese than patients with NG. Patients with CH had higher diastolic blood pressure in comparison with EH patients (p < 0.05). The main predictor of relative wall thickness (RWT) was the triglycerides to high density lipoprotein cholesterol (TG/HDL) ratio (R(2 ) = 0.319, beta = 0.246, p = 0.004). Patients received 12 months of antihypertensive treatment, either lifestyle modification only (n = 37) or lifestyle modification plus antihypertensive medications (n = 49) if severe ambulatory hypertension or target organ damage were present. After 12 months of treatment the prevalence of EH (37.2% vs 18.6%, p = 0.003) decreased but prevalence of CH did not change. Patients in whom RWT decreased also decreased waist circumference and TG/HDL; the main predictor of RWT decrease was a decrease of the TG/HDL ratio (beta = 0.496, R (2) = 0.329, p = 0.002). In adolescents with PH, LV geometry is related to central obesity and insulin resistance. Decrease of abdominal obesity and insulin resistance are the most important predictors of normalization of LV geometry, however CH has lower potential to normalize LV geometry. PMID- 21626454 TI - Investigation on the phenolic constituents in Hamamelis virginiana leaves by HPLC DAD and LC-MS/MS. AB - Aqueous and acetone/water extracts from Hamamelis virginiana leaves were investigated to obtain a thorough insight into their phenolic composition. To secure compound integrity, a gentle extraction method including the exclusion of light was used. Liquid chromatography tandem mass spectrometry (LC-MS/MS) analyses yielded a fingerprint including 27 phenolic constituents. Quantification of the key compounds on an equivalent basis by high-performance liquid chromatography diode-array detection (HPLC-DAD) showed that gallotannins consisting of six to 11 galloyl units constitute the main fraction, whereas procyanidins and catechin represented only a minor part. Closer inspection revealed that both extracts possess virtually the same galloyl hexose distribution, and the octagalloyl hexose represents the major tannin constituent. Additionally, eight flavonol glycosides and their corresponding aglycones quercetin and kaempferol, as well as three chlorogenic acid isomers and other hydroxycinnamic acids, were identified. Moreover, stability studies on the aqueous extract (5 degrees C, dark; room temperature, dark; room temperature, light) revealed that the phenolic profile underwent changes when exposed to light. Especially the gallotannins proved to be considerably unstable which may result in phytochemically altered Hamamelis leaf extracts upon transport and storage. PMID- 21626455 TI - The tumor necrosis factor type 2 receptor plays a protective role in tumor necrosis factor-alpha-induced bone resorption lacunae on mouse calvariae. AB - Tumor necrosis factor (TNF)-alpha exerts its biological function via TNF type 1 and type 2 receptors (TNFR1 and TNFR2). We have previously reported that bone resorption induced by lipopolysaccharide (LPS) in TNFR2-deficient mice is accelerated compared to that in wild-type (WT) mice. Although these results suggested that TNFR2 might have a protective role in bone resorption, we could not exclude the possibility that TNFR2 has no role in bone resorption. To clarify the role of TNFR2, we developed a TNF-alpha-induced bone resorption model using cholesterol-bearing pullulan nanogel as a TNF-alpha carrier to minimize the influence of inflammatory cytokines other than TNF-alpha. Injections of human TNF alpha (hTNF), an agonist of mouse TNFR1, stimulated bone resorption lacunae on the calvariae in WT mice, but mouse TNF-alpha (mTNF), an agonist of both mouse TNFR1 and TNFR2, could not. To eliminate the possibility that the TNFR1 agonistic effects of hTNF were stronger than those of mTNF, we used the same model in TNFR2 deficient mice. Injection of mTNF resulted in clear bone resorption lacunae to the same extent observed after using hTNF in the TNFR2-deficient mice. Histomorphometric analysis of osteoclast number supported the observed changes in bone resorption lacunae. These data suggest that TNFR2 has a protective role in TNF-alpha-induced bone resorption. PMID- 21626456 TI - Establishment of a simple test for iron absorption from the gastrointestinal tract. AB - Recent studies on iron metabolism have begun to reveal the molecular mechanisms underlying iron absorption, which is dramatically affected in several disorders. In the clinical setting, the ability to determine the status of iron absorption would aid in the diagnosis of pathological conditions. Efforts to develop an oral iron absorption test (OIAT) date back to at least 60 years. However, previous procedures were associated with a number of problems, such as radiation exposure and low reproducibility. In an attempt to resolve these issues, we employed sodium ferrous citrate (SFC), by which the influence of various nutrients and drugs on iron absorption was markedly reduced. We found that OIAT using SFC was simple to perform in both hospitalized patients and outpatients. The increment of serum iron and % transferrin saturation at 120 min after SFC administration was useful in distinguishing iron absorption between healthy volunteers, patients with iron-deficiency anemia, and patients with anemia secondary to chronic disorders, which are respectively characterized by normal, enhanced, and reduced iron absorption. Thus, the SFC-based OIAT may represent a viable screening test for the evaluation of gastrointestinal iron absorption. PMID- 21626457 TI - Current molecular markers for gastric progenitor cells and gastric cancer stem cells. AB - Gastric stem and progenitor cells (GPC) play key roles in the homeostatic renewal of gastric glands and are instrumental in epithelial repair after injury. Until very recently, the existence of GPC could only be inferred by indirect labeling strategies. The last few years have seen significant progress in the identification of biomarkers that allow prospective identification of GPC. The analysis of these unique cell populations is providing new insights into the molecular underpinnings of gastric epithelial homeostasis and repair. Of closely related interest is the potential to identify so-called cancer stem cells, a rare subpopulation of tumor-initiating cells. Here, we review the current useful biomarkers for GPC, including: (a) those that have been demonstrated by lineage tracing to give rise to all gastric cell lineages (e.g., the villin-transgene marker as well as Lgr5); (b) those that give rise to a subset of gastric lineages (e.g., TFF2); (c) markers that recognize cryptic progenitors for metaplasia (e.g., MIST1), and (d) markers that have not yet been analyzed by lineage tracing (e.g., DCKL1/DCAMKL1, CD133/PROM1, and CD44). The study of these markers has been mostly limited to the mouse model, but the hope is that the rapid pace of recent breakthroughs in this animal model will soon lead to a greater understanding of human gastric stem cell biology and to new insights into gastric cancer, the second leading cause of cancer-related death worldwide. PMID- 21626458 TI - Corporate social responsibility for nanotechnology oversight. AB - Growing public concern and uncertainties surrounding emerging technologies suggest the need for socially-responsible behavior of companies in the development and implementation of oversight systems for them. In this paper, we argue that corporate social responsibility (CSR) is an important aspect of nanotechnology oversight given the role of trust in shaping public attitudes about nanotechnology and the lack of data about the health and environmental risks of nanoproducts. We argue that CSR is strengthened by the adoption of stakeholder-driven models and attention to moral principles in policies and programs. In this context, we examine drivers of CSR, contextual and leadership factors that influence CSR, and strategies for CSR. To illustrate these concepts, we discuss existing cases of CSR-like behavior in nanotechnology companies, and then provide examples of how companies producing nanomedicines can exhibit morally-driven CSR behavior. PMID- 21626459 TI - Educational paper: imaging child abuse: the bare bones. AB - Fractures are reported to be the second most common findings in child abuse, after skin lesions such as bruises and contusions. This makes careful interpretation of childhood fractures in relation to the provided clinical history important. In this literature review, we address imaging techniques and the prevailing protocols as well as fractures, frequently seen in child abuse, and the differential diagnosis of these fractures. The use of a standardised protocol in radiological imaging is stressed, as adherence to the international guidelines has been consistently poor. As fractures are a relatively common finding in childhood and interpretation is sometimes difficult, involvement of a paediatric radiologist is important if not essential. Adherence to international guidelines necessitates review by experts and is therefore mandatory. As in all clinical differential diagnoses, liaison between paediatricians and paediatric radiologists in order to obtain additional clinical information or even better having joint review of radiological studies will improve diagnostic accuracy. It is fundamental to keep in mind that the diagnosis of child abuse can never be solely based on radiological imaging but always on a combination of clinical, investigative and social findings. The quality and interpretation, preferably by a paediatric radiologist, of radiographs is essential in reaching a correct diagnosis in cases of suspected child abuse. PMID- 21626460 TI - [Work related medical rehabilitation, MBOR - a process model in medical rehabilitation]. PMID- 21626461 TI - [Success factors of work-related orthopaedic rehabilitation]. AB - Work-related rehabilitation has for several years been gaining greater importance in orthopaedic rehabilitation. High-quality studies have confirmed that work related medical rehabilitation has favourable effects on earning capacity and work-life participation. This does however not hold true for all work-related rehabilitation programmes. In this context, 5 theses concerning success factors of work-related medical rehabilitation are developed. It is set out that the effects of work-related medical rehabilitation on work ability and work-life participation can be improved if programmes realize a needs-oriented assignment, include cognitive-behavioural components, follow a multimodal approach, step up treatment intensity, and if treatments are manualized. PMID- 21626462 TI - [Vocational orientation in medical rehabilitation: development of a practice handbook and a homepage as user-oriented media for rehabilitative practice]. AB - Vocational orientation in medical rehabilitation has become an important issue in rehabilitation science and practice in Germany. Although a variety of vocationally oriented interventions has been developed in recent years, there is still a lack of consensus with regard to the definition of specific types of interventions as well as relevant criteria (e. g., methods; instruments; duration/frequency; patient groups). Building on preliminary definitions, basic types of vocationally oriented measures/interventions were conceptually modified and refined in the context of a consensual process. 39 experts of various professions were contacted during a 2-stage Delphi survey to evaluate existing definitions of vocationally oriented interventions using key questions. Standardized descriptions of 5 vocationally oriented basic interventions were specified in cooperation with an interdisciplinary panel of experts. Descriptions were published in a workbook and a homepage for researchers and clinicians, which also contain information on the implementation of measures into rehabilitative care as well as good practice examples. The media developed in this project may contribute to the transfer of research results on vocationally oriented rehabilitation into health care practice. PMID- 21626463 TI - [A profile of requirements for the performance of work related medical rehabilitation from the perspective of the statutory pension insurance]. AB - OBJECTIVES: Work-related medical rehabilitation (Work MR) is a specific part of medical rehabilitation, which is well-established and broadly investigated. In almost all indications there is evidence of benefit of Work MR in patients with distinctive work-related problems due to chronic disease or disability. Nevertheless, there is a lack of implementation in clinical practice. This is due to a variety of reasons, mainly a missing definition of the appropriate Work MR interventions. The development of a Profile of Requirements (PoR) for defining contents and patterns of Work MR on behalf of the Statutory Pension Insurance (SPI) therefore is a fundamental step towards evidence-based, needs-focused rehabilitation management. The article introduces such a PoR - for the present focused on somatic indications. METHODS: The PoR was developed by an SPI expert group and revised in an extensive discourse within the SPI. A strategy involved in this context was the dissemination of work-related therapeutical concepts in medical rehabilitation. RESULTS: In the PoR, Work MR is defined as a concrete work-related designing of therapeutical interventions as well as an integrated "philosophy" in thinking and practicing of the actors involved. Work MR is differentiated in 3 intervention levels: While work-related basic interventions (Level A) are provided in all facilities for all patients, core interventions (Level B) as well as specific interventions (Level C) are explicitly concentrated on patient subgroups with distinct work-related problems. In the PoR the relevant diagnostic and therapeutic work-related interventions are attributed to the different intervention levels and described concerning the concrete arrangement. The 3 central core interventions "Work-related psychological group interventions", "Work-related internal load test" and "Work conditioning" are presented. CONCLUSIONS: The decisive benefit of the PoR is to substantiate the requirements of Work MR on behalf of the SPI often worded in a rather unspecific manner in the past. In the current pilot study called "Work MR Management" the feasibility of Work MR in clinical practice based on the PoR is tested. The resultant recommendations will provide the basis for disseminating Work MR throughout Germany independent of setting and indication. PMID- 21626464 TI - [Evidence based standards for rehabilitative therapy and work-related medical rehabilitation: are they compatible?: an expenditure analysis of treatment requirements]. AB - BACKGROUND: Medical rehabilitation carried out by the German Pension Insurance aims at maintaining or restoring the ability to work taking into account individual problems of the insured population. To ensure high-quality rehabilitation the German Pension Insurance uses a variety of quality assurance instruments. It is indispensable that requirements from different instruments constitute a conclusive and hence reproducible overall concept. This article looks at the requirements imposed on work-related medical rehabilitation (MBOR) and from the standards for rehabilitative therapy. METHODS: In an expenditure analysis we evaluated the extent to which the requirements from our standards for rehabilitative therapy in chronic back pain and work-related medical rehabilitation are compatible with each other, and their impact on the quantity of treatment to be delivered. In a first step the instruments were compared qualitatively. Next, we looked at the resulting cumulative duration of treatment. Finally, using an orthopaedic rehabilitation centre on the underlying assumptions as an example, we analysed whether existing staffing levels are sufficient to fulfil the requirements. RESULTS: MBOR and rehabilitation treatment standards both set requirements regarding vocational orientation; they, however, employ different methods and address different groups of rehabilitants. The duration of treatment for a rehabilitant who - owing to his work-related problems - is treated not only according to the rehabilitative treatment standards but also fulfilling the MBOR requirements profile increases on average from 10.4 to 14.2 h a week. Modelling of the staffing levels necessary to fulfil the requirements shows a possible shortage in the group of social workers only. DISCUSSION: The requirements from MBOR and rehabilitative treatment standards are compatible with each other regarding systematics, target group and scope of work-related treatment and are consistent with specifications for structural quality, especially with staffing levels. Rehabilitants are not exposed to undue strains, and staffing levels in rehabilitation centres are adequate - with the possible exception of social workers for MBOR. In future development of quality specifications for rehabilitation it remains pre-eminent to ensure an interlocking of the different requirements, in order to provide the rehabilitation centres with a comprehensible frame for the provision of care. PMID- 21626465 TI - [Who benefits from stepwise occupational reintegration provided under the statutory pension insurance scheme?]. AB - BACKGROUND: Stepwise occupational reintegration (SOR) - since law amendments in April 2004 also provided under the German pension insurance scheme (Deutsche Rentenversicherung, DRV) - is an instrument intended to support insurants on sick leave in reintegrating into work step by step after long-term illness. In 2008, the effectiveness of SOR regarding return to work was affirmed for the first time in a comprehensive study. However, in view of the growing amount of SOR, the question of differential effects of SOR in special subgroups is raised. METHODS: This paper presents a re-analysis of data collected in the 2008 study. A total of 696 patients after medical rehabilitation were included in the analyses, 348 with SOR provided by the DRV, and a control group of 348 patients without SOR matched on a multitude of different variables using the Propensity Scores. Successful outcome was measured using a combined criterion "Return to work in good health", that is, patients returning to gainful activity and with sick leave of under 6 weeks and no intention to retire within a one-year follow-period after medical rehabilitation. Differentiating criteria are age gender, rehab indication, periods of sick leave in the year before medical rehabilitation, kind of and access to medical rehabilitation. RESULTS: The data indicate especially good results of SOR for patients with mental disorders (OR=2.49), patients who were requested to participate in medical rehabilitation by a health insurance fund because of long-term sick leave (OR=2.71), and patients with longer periods of sick leave before medical rehabilitation (3 to <6 months: OR=2.41, 6 months and more: OR=2.23). In contrast, there are only minimal effects (statistically not significant) of SOR in patients with medical rehabilitation directly after a hospital stay ("Anschlussheilbehandlung"), patients with cardiac or oncological diseases, and in younger (age 19-34) and older patients (age 55-60). In-depth analyses show that SOR success is more marked in patients with poorer return to work prospects. DISCUSSION: The findings indicate differential effects of SOR after medical rehabilitation for subgroups, effects associated in particular with return to work problems, kind of disease, and age. There is evidence for greater benefits of SOR in groups of patients with a high risk of non-successful reintegration. Hence, SOR opens up new options after medical rehabilitation in patients with especially severe impairments. PMID- 21626466 TI - [Intensified rehabilitation aftercare (IRENA): utilization alongside work and changes in work-related parameters]. AB - BACKGROUND: An aftercare programme following medical rehabilitation may be beneficial in order to reinforce and stabilize the positive effects of rehabilitation and to encourage individual health-related modifications of behaviour and lifestyle. Medical rehabilitation and the aftercare programme of the German Pension Insurance Fund primarily are intended to sustain earning capacity. As part of an evaluation of the Intensified Rehabilitation Aftercare Programme (IRENA) established by the German Pension Insurance Fund, work-related aspects in orthopaedic patients were analyzed based on various data sources. Firstly, the significance of institutional and individual conditions for utilization of IRENA alongside work was of interest. Secondly, the IRENA participants' judgements of the changes of work-related parameters due to the programme were examined, differentiating specifically by extent of earning capacity impairments as well as by particular work problems. METHODS: The data set used for the analysis is composed of person-related routine data of the German Pension Insurance Fund relative to IRENA records of the year 2007 (n=30 663), interview data from orthopaedic rehabilitation centres providing IRENA (n=225), and questionnaires of IRENA participants (n=750) that were either collected during a broad evaluation of the IRENA programme or provided by the German Pension Insurance Fund. RESULTS: The results show that the compatibility of IRENA and work is facilitated by the institutional conditions. However, differences between inpatient and outpatient settings have to be recognized. The possibilities to participate in IRENA throughout the day frequently are more diverse in an outpatient setting. In contrast to inpatient centres, outpatient rehabilitation centres see clearly better chances for patients to return to work and to participate in IRENA alongside. With respect to the work-related parameters (work ability, periods of sick leave), clear improvements were reported by participants from the start of rehabilitation to the survey time after the end of IRENA. Particular work problems were reported by 33% of the IRENA participants. The work ability at the end of rehabilitation was found to have been the essential factor for improvement of work ability following IRENA. Particular work problems, however, had no influence, these individuals profited from IRENA to an equal extent. CONCLUSIONS: Institutional and individual view show that IRENA is compatible with utilization alongside work. Also, IRENA combined with prior medical rehabilitation will bring about subjective improvements in health and work-related parameters. PMID- 21626467 TI - [Adaptive testing in rehabilitation: a way to economic assessment of patients' attributes]. AB - For diagnostics and outcome measurement in clinical rehabilitation a multitude of questionnaires is used. In order to gain comparability of the diagnostic findings, generally, the same information is gathered of all patients, regardless of their state of health or how severely ill they are, by using identical groups of items. In this kind of assessment it is, however, problematic that (a) the assessment instrument usually only allows for adequate and reliable diagnostics of patients who suffer from injuries or illnesses of medium severity, and (b) that an uneconomic data collection is required because of an extensive set of items which may in individual cases be uninformative and unnecessary. This article shows how data assessment can be adjusted to the individual handicaps of the patient by using adaptive or tailored testing. Thus it can be ensured that the testing becomes more economical and results more precise. Furthermore, a high acceptance by the patients participating in the data collection can be achieved, as the test items submitted fit the ability of the patients adequately. Yet, adaptive test systems require a complex and very careful psychometrical development of the assessment procedure in such a way that in spite of presenting different items to patients, diagnostic outcomes are comparable between patients and for different points in time (e. g., pre-post comparison). This article shows the steps of development that have to be accomplished in order to set up a psychometrically sound adaptive test procedure. Moreover, it shows how its usage in the area of medical rehabilitation can be profitable for handlers and patients. PMID- 21626468 TI - [Influence on flux density of intraoral dental magnets during 1.5 and 3.0 tesla MRI]. AB - PURPOSE: When using dental duo-magnet systems, a mini-magnet remains in the jaw after removal of the prosthesis. In some cases, implant-borne magnets may be removed, whereas tooth-borne magnets are irreversibly fixed on a natural tooth root. The goal of this paper is to identify the impacts of the duration and orientation of exposure on these magnets in a 1.5 or 3 Tesla MRI. MATERIALS AND METHODS: For this study, 30 SmCo and 60 NdFeB magnets were used. During the first experiment, they were exposed with free orientation for 64 minutes. During the second experiment, the magnets were fixed in position and exposed at 1.5 and 3 Tesla while aligned in a parallel or antiparallel direction. RESULTS: While the duration of exposure in MRI is irrelevant, the orientation is not. The coercive field strength of these NdFeB and SmCo alloys is not sufficient to reliably withstand demagnetization in a 1.5 or 3 T MRI when aligned in an antiparallel direction. At 1.5 T neodymium magnets were reduced to approx. 34 % and samarium magnets to approx. 92 % of their initial values. At 3 T all magnets were reversed. CONCLUSION: As a precaution, the worst-case scenario, i. e. an antiparallel orientation, should be assumed when using a duo-magnet system. If an MRI can be postponed, the general dentist should remove implant-borne magnets. If there is a vital indication, irreversible damage to the magnets is acceptable in consultation with the patient since the replacement costs are irrelevant given the underlying disease. PMID- 21626469 TI - [Emotional childhood parentification and mental disorders in adulthood]. AB - BACKGROUND: Emotional parentification is considered harmful to a child's development. METHOD: A total of about 975 patients were examined at a Department of Psychosomatic Medicine and in the practices of general practitioners with regard to childhood adversities. RESULTS: Emotional parentification is a risk factor for 2 symptom groups: the patients with depression and the patients with somatoform pain. While the occurrence of depression is mainly predicted by maternal emotional parentification, paternal influences are also relevant in regard to the development of somatoform pain. CONCLUSION: Emotional parentification is an important risk factor for the occurrence of psychological and somatoform complaints in adulthood. This is especially apparent in combination with further risk factors, such as low reported values for love, sexual abuse, or being raised without a father. PMID- 21626470 TI - Oncolytic viruses for the treatment of neuroendocrine tumors. AB - Oncolytic viruses are emerging as anticancer agents, and they have also shown great promise for use against neuroendocrine tumors. Many viruses have a natural tropism for replication in tumor cells. Others can be genetically engineered to selectively kill tumor cells. Viruses have some advantages as therapeutic agents over current cytotoxic drugs and small molecules. They replicate in tumor cells and thereby increase in number over time leading to increased dosage. They are immunogenic and can alter the immunosuppressive tumor microenvironment and activate immune effector cells. They have also been shown to be able to kill drug resistant cancer stem cells. This article reviews the recent literature on oncolytic viruses used so far for neuroendocrine tumors and indicates important issues to focus on in the future. PMID- 21626471 TI - Effects of adiponectin in TNF-alpha, IL-6, and IL-10 cytokine production from coronary artery disease macrophages. AB - Adiponectin, an adipose tissue secreted protein, exhibits anti-inflammatory and antiatherogenic properties. We examined the effects of the globular and full length adiponectin on cytokine production in macrophages derived from Coronary Artery Disease (CAD) patients and control individuals. Adiponectin's effects in human macrophages upon lipopolysaccharide (LPS) treatment were also examined. Full length adiponectin acted differently on TNF-alpha and IL-6 production by upregulating TNF-alpha and IL-6 protein production, but not their mRNA expression. Additionally, full length adiponectin was unable to abrogate LPS proinflammatory effect in TNF-alpha and IL-6 mRNA expression in CAD and NON-CAD macrophages. In contrast, globular adiponectin appeared to have proinflammatory properties by potently upregulating TNF-alpha and IL-6 mRNA and protein secretion in human macrophages while subsequently rendered cells resistant to further proinflammatory stimuli. Moreover, both forms of adiponectin powerfully suppressed scavenger MSR-AI mRNA expression and augmented IL-10 protein release, both occurring independently of the presence of LPS or CAD. These data indicate that adiponectin could potentially protect human macrophages via the elevated IL 10 secretion and the suppression of MSR-AI expression. It can also be protective in CAD patients since the reduced adiponectin-induced IL-6 release in CAD macrophages compared to controls, could be beneficial in the development of inflammation related atherosclerosis. PMID- 21626472 TI - Risk of hyperamylasemia and acute pancreatitis after double-balloon enteroscopy: a prospective study. AB - BACKGROUND AND STUDY AIMS: There have been reports, mainly retrospective, of pancreatitis and hyperamylasemia after anterograde double-balloon enteroscopy (DBE). Our aim was to report the incidence of pancreatitis and hyperamylasemia after DBE and investigate possible risk factors associated with its occurrence. PATIENTS AND METHODS: In this single-center prospective cohort study, serum samples were taken for amylase and lipase before and 3 hours after anterograde DBE in consecutive patients. Multiple variables were recorded, including total procedure time, insertion depth, and number of passes. Patients were evaluated to 24 hours later for signs of pancreatitis. The main outcome measures were the occurrence of hyperamylasemia and pancreatitis. RESULTS: 92 patients were included in the analysis (58 women, 34 men; mean age 54 years, range 18-89). The mean total procedure time was 62 minutes (range 30-120). The mean post-procedure amylase and lipase levels were significantly higher in comparison with the baseline levels (165 U/L vs. 69 U/L and 144 U/L vs. 28 U/L respectively, P<.05); 36 patients (39%) showed hyperamylasemia after the procedure and three patients developed acute mild pancreatitis. Hyperamylasemia was associated more frequently with procedure duration greater than 60 minutes ( P<.001) and insertion depth greater than 25 cm ( P<.013). CONCLUSIONS: The incidence of hyperamylasemia after anterograde DBE is common and particularly associated with longer procedure time and insertion depth. The cumulative incidence of pancreatitis was 3%. We recommend the avoidance of both unnecessarily lengthy procedures and deep insertion distances in patients who undergo anterograde DBE. PMID- 21626473 TI - [Otologic diagnosis: powder technique]. AB - The otologic diagnostic powder test with grape - sugar simple will be performed with a powder - blower. It permits a good differentiation between dry squamous epithelium and wet mucosa. In our opinion this test therefore is unrenouncable in microscopic differentiation between mucosa in central perforations on one hand and atrophic scars, retraction pockets or adhesive epithelium of the drum or complete atelectasis of the tympanic cavity on the other hand. PMID- 21626474 TI - [Health valuations for patients with chronic ischemic heart disease]. AB - OBJECTIVE OF THE STUDY: The study deals with the question of how patients with chronic ischemic heart disease assess different health situations that can be achieved by rehabilitation. Furthermore it examines which factors influence these health valuations and whether the predictors vary depending on the level of education. METHODS: The health valuations of n = 331 patients with chronic ischemic heart disease are compiled using visual analogue scales (VAS). In addition to sociodemographic questions, generic and illness-specific scales (SF 12, MacNew) for the health-related quality of life (HRQOL) are used as potential predictors of the health valuations. Additional basic medical data were provided by the physician. Hierarchical regression analyses are conducted; the sociodemographic, medical and HRQOL variables are included stepwise. Since many variables are observed for the regression models, an imputation of missing values is made. RESULTS: The health dimensions "Self-care and domestic life" and "Mobility" are assigned the highest values on the VAS. The lowest preference is assigned to the dimensions "Reduction of symptoms" and "Information about the disease". The differences between the health dimensions are statistically significant. Sociodemographic variables explain up to 3.6% of the variance of health valuations, with level of education and living with a partner being the most important predictors. The medical variables included in the second step explain between 2.1 and 6.8% incremental variance; the most important predictor is the operation performed prior to rehabilitation (bypass, heart valve). The HRQOL variables in the third block provide 7.1-24.9% incremental explanation of variance, by far the highest percentage. This is mainly achieved using the 3 MacNew scales (emotional, social and physical functioning). The overall explanation of variance for the health valuations is 17.1-28.8%. For patients with a higher level of education, the total explanation of variance is about 9.2% higher on average than for the total sample and lies between 21.2 and 44.1%. DISCUSSION: The health dimensions examined are assessed quite differently by the patients. The most significant factor of influence is the HRQOL. In the subgroup of patients with a higher level of education, the predictive strength of the variables examined is considerably higher in comparison with the total sample. Since the health valuations can be predicted to a limited extent only using other data arising during routine care, it is necessary to implement special methods for compiling health valuations. PMID- 21626475 TI - The Early Rehabilitation Barthel Index (ERBI). AB - An extended version of the Barthel index, the so-called "Early Rehabilitation Barthel Index", or ERBI, is widely used in German early neurological rehabilitation centres and even was used in defining early rehabilitation procedures in the German DRG system. It contains highly relevant items like mechanical ventilation, tracheostomy, or dysphagia. This study presents an English version of the ERBI and examines its validity and reliability. Two samples of early neurological rehabilitation patients have been analyzed. In one sample (n=1,669), measures of morbidity and length of stay (LOS) have been compared between certain ERBI categories. In a second sample (n=273), inter-rater reliability (nurses vs. physicians) has been examined. Patients with low ERBI had a significantly longer LOS than those with high ERBI values (p<0.001). Further, parameters of morbidity (patient clinical complexity level, number of co diagnoses) were significantly higher in a low ERBI subgroup. Inter-rater reliability was r=0.849 (p<0.001). The findings suggest that the ERBI is a reliable and valid scale to assess early neurological rehabilitation patients. PMID- 21626476 TI - [Operationalization and feasibility of the early rehabilitation index ("lower saxony early rehabilitation index")]. AB - This paper suggests an operationalization of the items of the early rehabilitation Barthel-index (ERBI). In addition, it presents a modification of this instrument, called lower-saxony ERBI (lsERBI). In order to validate the lsERBI, it has been used in 273 neurological rehabilitants. Findings suggest that the lsERBI has a higher specificity but lower sensitivity than the conventional ERBI. Inter-rater-reliability was high (r=0.84). Further studies are welcome to validate the lsERBI. PMID- 21626477 TI - [Medically unexplained and somatoform complaints and disorders in the elderly: a systematic review of the literature]. AB - OBJECTIVE: To provide a systematic overview of the research concerning the frequency of medically unexplained, somatoform complaints and disorders in the elderly, their risk factors, comorbidity, course, management in primary and secondary care, and psychotherapy. METHOD: We evaluated 248 clinical and randomized trials, reviews, meta-analyses and practice guidelines for adult samples identified from "PubMed" and "PsycInfo" using the search terms "somatoform" OR "medically unexplained" OR "somatization" OR "somatisation" in combination with other terms for their relevance for the elderly. Other relevant trials were identified from the references from these publications. RESULTS: Medically unexplained, somatoform complaints and disorders occur in older persons and are more frequent in elderly women than in elderly men. Although many studies found no increased frequency of somatoform complaints in the elderly, based on present research we cannot draw final conclusions concerning the frequency of somatoform disorders and complaints in the elderly, and we cannot give evidence based recommendations for their treatment in primary and secondary care and psychotherapy. DISCUSSION: This situation results from the special problems encountered in the diagnosis of somatoform complaints and disorders in the elderly, from problems in conceptualisation also found in younger adults, and from the lack of research concerning treatment focussed solely on the old and very old. PMID- 21626478 TI - [Surviving the initial phase: subjective theories of illness in patients suffering from acute leukaemia at the end of initial inpatient treatment]. AB - OBJECTIVES: Studies concentrating on the temporal dependence of subjective concepts during oncological treatment are underrepresented. Subjective interpretation contexts develop in the course of illness. The study focuses on the ideal-typical gestalt of these contents. METHODS: In a follow-up study on coping, 12 patients with acute leukaemia (AL) were interviewed using a semistructured interview at the end of initial inpatient treatment. Using qualitative methodology, we inductively developed categories and assigned them to formal main categories. RESULTS: The following categories were developed: causal uncertainty as burden; discrepancy between subjective and objective assessment of degree of threat; knowledge of disease: conflict between information-seeking and information-avoiding behavior; dominance of medical approach to treatment; pursuit of normality; defense of emotions; orientation to workflows on the ward; adjustment as a coping strategy; positive attitude as a resource; life between hope and fear; limited future; latent fear of death. CONCLUSION: Themes of coping with the disease become visible. Some of these contents are tacit and latent, although of high subjective relevance to the patient. Their consideration could improve the patient-physician relationship. PMID- 21626479 TI - [Patients' perspectives on behavioral and cognitive changes after hospitalization: a 3-month follow-up cohort study using qualitative and quantitative research methods]. AB - OBJECTIVES: To describe patients' subjective perspectives on hospitalization at time of admission, to prospectively investigate patients' views on changes actually achieved after discharge, and to identify predictors of subjective changes 3 months after hospitalization. METHODS: We conducted a 3-month follow-up cohort study using qualitative and quantitative research methods. Consecutive adult inpatients of an internal medicine ward and a ward integrating internal and psychosomatic medicine were included. They answered open-ended questions concerning their motivation for change at the time of admission (N = 639) using self-report questionnaires and 3 months post hospitalization via a telephone interview (N = 389). Qualitative content analysis was performed. Predictors of subjective change 3 months after hospitalization were identified using multivariate logistic regression analyses. RESULTS: At admission, more psychosomatic patients than internal-medicine ward patients wanted to achieve change (58% vs. 44 %). Patients expressed desires that were assigned to three main categories: to pay more attention to one's physical health (40 %), to change one's perceptions (32 %), and to change one's way of life (16 %). Three months after discharge, 52 % of patients had experienced cognitive and behavioral changes they attributed to their hospitalization. 31 % of the patients reported continuing to implement changes that had begun during hospitalization. Predictors of subjective change 3 months after hospitalization included high educational level (OR = 2.04, 95 %-CI 1.27-3.26), young age (OR = 0.98, 95 %-CI 0.97-1.00), and length of hospitalization (OR = 1.03, 95 %-CI 1.00-1.05). CONCLUSIONS: There is considerable motivation for behavioral and cognitive change among inpatients following admission; this deserves the attention of healthcare professionals. An integrated psychosomatic setting would appear to support cognitive and behavioral changes more effectively than a classical internal medicine ward. PMID- 21626480 TI - [Age-, gender- and diagnosis-specific changes of body concept during stationary psychosomatic treatment]. AB - OBJECTIVE: This study investigates body concept with respect to psychiatric diagnosis, age and gender of inpatients before and after treatment. METHODS/SAMPLE: We examined 153 inpatients with 131 complete datasets. The examination was done at the beginning and at the end of the treatment using the Frankfurt Body Concept Scale (FKKS, Deusinger 1998). The results were calculated by t-test for dependent samples, posthoc Scheffe-tests and univariate variance analysis. RESULTS: The diagnoses indicated a more positive body concept at the end of the treatment (patients with somatoform disorder had effect sizes between 0.2 and -0.4, patients with depressions between -0.4 and -0.7, and patients with eating disorders between -0.3 and -0.7). Gender and age had only little influence on the positive changes. DISCUSSION: The improvement of body concept through inpatient treatment represents a challenge especially for somatoform disorders. The duration of this improvement after inpatient treatment needs further investigation. PMID- 21626481 TI - [Impact of commuting on partnership and family life. A literature review of the current state of research with special emphasis on commuter marriages]. AB - Because of modern working conditions, for many people commuting has increasingly become a necessity with many consequences for their health as well as for their family life and partnership. To date, little research has been done concerning the impact of this modern lifestyle on partnership and family life, particularly on dual-career couples and long-distance commuters. The results of a literature review are presented based on age, sex, family cycle and consequences for children; research questions are formulated. Sex-specific differences seem to be especially important, confirming the "household responsibility hypothesis." The commuter marriage as a new lifestyle seems to have become a coping mechanism for the increasing necessity in our society to commute. PMID- 21626482 TI - [The clinical relevance of defense mechanisms in inpatients and outpatients]. AB - OBJECTIVES: How clearcut is the clinical significance of mature and immature defense mechanisms in psychological symptoms? METHODS: Defense mechanisms, symptoms and well-being were assessed in 293 inpatients, 316 outpatients and 157 students. RESULTS: The factors "mature defense" and "immature defense" predict well-being/symptoms to a substantial degree. DISCUSSION: The assessment of defense mechanism via self-report is clinically useful. PMID- 21626483 TI - [The kidney transplant recipient]. PMID- 21626484 TI - [Piedmont: a region with a difference]. PMID- 21626485 TI - [Rare genomic variants and susceptibility to multifactorial diseases. The example of medullary sponge kidney]. PMID- 21626486 TI - [ER stress: a pivotal role for TNF-alpha]. PMID- 21626487 TI - [Malnutrition-inflammation syndrome in dialysis: in search of new biomarkers]. PMID- 21626488 TI - [Early hematopoietic response to ESA: an important prognostic factor?]. PMID- 21626489 TI - [Medical radiation exposure in dialyzed patients]. PMID- 21626490 TI - [Loop diuretics in acutely decompensated heart failure: high doses absolved but still doubts about continuous infusion?]. PMID- 21626491 TI - [Proteinuria: a risk factor not to be overlooked in SLE]. PMID- 21626492 TI - [Arterial hypertension in dialysis: up to what point should it be corrected? A lot!]. AB - Arterial hypertension is a powerful predictor of cardiovascular events and mortality in the general population. It is well known that the prevalence of hypertension is high in hemodialysis patients, but there is still debate on the existence of a cause-effect relationship between high blood pressure and mortality in patients with end-stage renal disease. Although some authors have suggested a positive association between low blood pressure values and mortality, various studies have shown that blood pressure reduction improves survival and reduces cardiovascular events in hemodialysis patients. Furthermore, a close relationship between blood pressure and mortality has been revealed recently by data obtained from ambulatory and home blood pressure monitoring, whereas blood pressure values recorded in dialysis units seem to have poor prognostic value. Even though a hemodialysis session generally induces a reduction in blood pressure, in part of the patients a rise in blood pressure may be observed. This phenomenon, known as intradialytic hypertension, is associated with a worse prognosis in terms of mortality and with an increased incidence of hospitalization. As the extracellular fluid level is the main factor influencing blood pressure values in hemodialysis patients, correct determination of dry weight is of fundamental importance in order to obtain blood volume and pressure control in this population. Regarding antihypertensive drug therapy, the few trials that are currently available have shown that angiotensin-II inhibitors and dihydropyridine calcium channel blockers exert a protective cardiovascular effect. In conclusion, high blood pressure levels in hemodialysis patients are associated with an increased risk of death and cardiovascular events. Reduction of blood pressure in these patients improves survival and reduces adverse events just like in hypertensive patients without end-stage renal disease. It is possible, however, that patients who have more comorbidities may not tolerate, especially during hemodialysis sessions, certain blood pressure levels that are known to be preferable in subjects that are less compromised. PMID- 21626493 TI - [Arterial hypertension in dialysis: up to what point should it be corrected? It depends]. AB - It is well known that blood pressure values <130/80 mmHg in high-risk patients (diabetics, heart patients) reduces the cardiovascular risk, and it is equally well known that the same target is advised for patients with renal insufficiency. In view of the elevated prevalence of postural hypotension, autonomic dysfunction and peripheral, cerebral and cardiac vasculopathy in kidney patients, the maintenance of this pressure target can, however, be harmful in this category of patients. Indeed, some studies have shown that lower pressure values are associated with more cardiovascular events in dialysis patients. The increased incidence of organ damage consequent to the hypotensive episodes that may occur in these patients should also be taken into account. Hypotension during dialysis, especially when occurring repeatedly, has indeed been indentified as the cause of the increased risk of intestinal, cardiac and cerebral ischemia in such patients. Antihypertensive treatment must therefore be seen in a different light compared with that of the general population, and we should consider the anatomic and physiopathological peculiarities of dialysis patients. It thus becomes of primary importance to assess the presence of comorbidities and cardiovascular risk factors that may favor the maintenance of pressure values that are not necessarily extremely low. A carefully reasoned use of antihypertensive drugs which takes these aspects into account could reduce the incidence of ischemic events with consequent organ damage that can increase dialysis patient mortality. PMID- 21626494 TI - [Arterial hypertension in dialysis: the need of a tailorized treatment]. AB - There is no doubt that arterial hypertension in dialysis patients as well as in patients without renal disease is deleterious for the cardiovascular system. Reducing the pressure values to 130/80 mmHg is what the guidelines suggest. The dialysis setting, however, presents a very different scenario from that of the general population. Dialysis per se is the most powerful of the pharmacological measures in blood pressure control. At the same time, dialysis is an interfering factor in pressure control and can even foster the onset of a perverse vicious circle with, in the same patient, the onset of severe intradialysis hypotension and hypertensive crises in the interdialysis period. An obstinate pursuit of normal pressure values may give rise to sudden reductions in the arterial pressure that can generate vascular damage in various districts (cerebral, cardiac, mesenteric). From these premises arises the need to individualize, as far as possible, not only the pharmacological therapy but also the dialysis prescription in ways that will avert hypotension during the session and at the same time correct the chronic water and salt overload that occurs in the dialysis patient. PMID- 21626495 TI - [Opioid analgesics in patients with chronic renal failure: principles for use and current guidelines]. AB - The treatment of pain in patients with impaired renal function may be problematic, especially when opioid drugs need to be used. In the presence of renal failure, significant changes occur in the metabolism and pharmacokinetics of these drugs, which can lead to adverse reactions due to the accumulation of parental compounds and active metabolites. This paper presents and discusses the available evidence concerning the optimal use of the most frequently employed opioid analgesics to treat pain in patients with renal impairment. PMID- 21626496 TI - [The impact of known and unknown dietary components to phosphorus intake]. AB - The pathogenesis of CKD-MBD is multifactorial but the tendency towards phosphorus retention due to an excessive dietary intake for the residual renal function plays a central role. The dietary phosphorus is absorbed in the intestine as inorganic free phosphorus. The share of intestinal absorption (about 60% on average) is negligible for plant phosphorus (in the form of phytate), while it is maximal for phosphate or polyphosphates contained in food additives. The latter represent a dangerous extra load of phosphorus because they are poorly recognized by patients and widely used in modern nutrition, in particular in low-cost food. In a free mixed diet, the phosphorus content is directly related to that of proteins. It follows that protein-rich foods are the main source of phosphorus. This is a favorable condition for CKD patients in conservative therapy when a low protein diet is implemented, while it represents a huge problem for dialysis patients, who need a high-protein diet. A simple and effective approach to reduce the load of dietary phosphorus without reducing protein intake is to educate patients to avoid foods high in phosphorus (cheese, egg yolk, nuts, etc.), and particularly those containing phosphorus additives. In addition, they should prefer boiling (resulting also in a decrease in sodium and potassium) to other methods of cooking. Counseling by a dietician is important for successful patient care. The dietician provides nutritional education, can help the patient with the choice of food, and may favor the adherence to dietary prescriptions, which is a crucial aspect in an integrated approach to CKD-MBD. PMID- 21626497 TI - [Bioincompatibility of acetate even at low concentrations]. AB - Over the last 20 years a large body of evidence has demonstrated that in chronic renal failure there is progressive chronic inflammation, which increases after the start of dialysis. In this phase a fundamental role is played by bioincompatibility reactions induced by contact with the different dialysis materials: membranes, plastic lines, dialysis fluids as well as contaminants present in water. Clinically evident symptoms induced by bioincompatibility reactions are usually taken into serious account by nephrologists, while more subtle chronic effects, noxious in the long term, are often underconsidered. Since the 1990s many efforts have been addressed to membrane improvement and water treatment, while there is still a lot to be done for better dialysates. Acetate dialysis is routinely used in only about 5% of patients worldwide but over 80% of patients are exposed to the lower acetate concentrations present in standard bicarbonate dialysate. These concentrations are not negligible and are able to induce chronic reactions mainly converging on the endothelium, stimulating and maintaining the atherogenesis process with important long-term implications for cardiovascular morbidity. This review presents and discusses the available data on the cellular and molecular effects induced by acetate, even at low concentrations. PMID- 21626498 TI - [Nephrotoxicity induced by chemotherapy]. AB - Abnormal renal function markedly influences the clinical management of cancer patients. While the neoplasm may cause renal damage in itself, the damage may be exacerbated by treatment with chemotherapeutic agents. Since many chemotherapeutic agents are metabolized and excreted through the kidneys, their use may represent a major risk factor for the development of renal abnormalities. Moreover, when renal failure is present before chemotherapy treatment, certain drugs need dose adjustments or are not indicated. Careful monitoring of renal function during chemotherapy is thus necessary and preventive measures should be adopted when possible to reduce the occurrence of renal dysfunction. PMID- 21626499 TI - [Hyponatremia: practical approach to diagnosis and treatment]. AB - Hyponatremia is one of the most frequent ion and water disorders. It is generally due to disproportionate renal water retention. Severe hyponatremia (<125 mEq/L) is associated with well-known clinical symptoms and manifestations. However, even mild reductions in sodium blood levels have been shown to be associated with increased mortality and with the risk of falls and fractures. The diagnosis of hyponatremia, although requiring simple clinical and laboratory tests, may be complex and difficult. Treatment should be tailored to the clinical manifestations and severity of hyponatremia. Vaptans may be used in euvolemic or hypervolemic hyponatremia and particularly in SIADH. PMID- 21626500 TI - [Immigrants and dialysis: a survey in Piedmont]. AB - The number of immigrants has been rapidly increasing in Italy in the last decade, with potentially profound effects on the national health care system. Yet, few data are available on the clinical and demographic features of these subjects, or on their need for nephrological care and dialysis treatment. A survey was conducted in 19 dialysis facilities of Piedmont (a northwestern Italian region) about immigrants on chronic dialysis treatment. Data on native country, administrative position, clinical and dialysis aspects were anonymously collected. Overall, 93 immigrant dialysis patients coming from 24 foreign countries were registered. Most of them were young (mean age 46?14 years) and on extracorporeal treatment (87%); late referral (38%) or starting dialysis in emergency (17%) were common modalities of presentation. Glomerular (33%) or unknown (31%) nephropathies were the most representative causes of end stage renal disease. No difference in incidence of HCV, HBV and HIV compared with native Italian patients was observed. Notably, more than 50% of the immigrant patients had low-level knowledge of Italian. As regards administrative position, 69% were regular foreign citizens, 19% were temporary foreign workers, and 9% had a residence permit. Our survey confirms the existence of a young immigrant population on dialysis in Piedmont, whose social and relational problems are more challenging than clinical aspects and call for new organizational models to manage this growing population on dialysis. PMID- 21626501 TI - [Cox regression analysis in epidemiological research]. PMID- 21626502 TI - [Role of the ultrasounds in the kidney biopsy: a case of postbiopsy hematoma linked to renal arteriovenous fistula]. AB - In the planning of a kidney biopsy procedure, ultrasound examination has a crucial role before percutaneous renal biopsy (PRB) in detecting renal abnormalities that could contraindicate the biopsy; during PRB as a method to locate the kidney; and after PRB to diagnose and monitor postbiopsy complications. The case of a 40-year-old woman who underwent ultrasound-guided PRB for urinary abnormalities is described. Careful renal assessment by ultrasonography before the kidney biopsy was not performed. The post-PRB ultrasound examination revealed a perinephric hematoma along with an arteriovenous fistula (AVF) at the lower pole of the biopsied kidney. Surprisingly, a later renal angiography showed AVFs in both kidneys and therefore a diagnosis of non-iatrogenic, idiopathic AVFs was made. Based on our unusual case report, we discuss the diagnostic approach and therapeutic strategies for renal AVFs and we emphasize the usefulness of ultrasound for initial pre-PRB evaluation. In addition, with respect to the post-biopsy hematoma, the advantages offered by ultrasound during and after the implementation of PRB are debated. Finally, the role of post-biopsy hematoma as a possible indicator of post-PRB complications is reported. PMID- 21626504 TI - [Interview with Prof. Francesco Locatelli: an international nephrologist of all seasons by Timio Mario]. PMID- 21626503 TI - [A case of arteriovenous fistula stenosis treated with percutaneous transluminal angioplasty]. AB - Arteriovenous fistula (AVF) is considered the gold standard for vascular access in hemodialysis, even in elderly patients. Malfunction of AVF is the first cause of hospitalization in the HD population, with a correlated increase in costs. For the monitoring and surveillance of fistulas, the 2006 K/DOQI workgroup recommends physical examination, direct flow measurement, Doppler ultrasonography and recirculation (by a non-urea-based dilutional method). We report the case of a 67 year-old woman affected by AVF stenosis. Combined AVF surveillance with recirculation and Doppler ultrasonography permitted early diagnosis and treatment with percutaneous angioplasty. PMID- 21626505 TI - The HOX gene network in hepatocellular carcinoma. AB - Liver organogenesis and cancerogenesis share common mechanisms. HOX genes control normal development, primary cellular processes and are characterized by a unique genomic network organization. Less is known about the involvement of HOX genes with liver cancerogenesis. The comparison of the HOX gene network expression between nontumorous livers and hepatocellular carcinomas (HCCs) highlights significant differences in the locus A HOX genes, located on chromosome 7, with a consistent overexpression of HOXA13 mRNA thus validating this gene deregulation as a feature of HCC. HOXA13 is a determinant of gut primordia and posterior body structures. Transcriptome analysis of HCC/nontumorous liver mRNAs, selected on the basis of HOXA13 overexpression, recognizes a set of deregulated genes. The matching of these genes with previously reported HCC transcriptome analysis identifies cell-cycle and nuclear pore-related HCC phenotype displaying poor prognosis. HOXA13 and HOXA7 homeoproteins share a consensus sequence that physically links eIF4E nuclear bodies acting on the export of specific mRNAs (c myc, FGF-2, vascular endothelial growth factor (VEGF), ornithine decarboxylase (ODC) and cyclin D1). We report the protein-protein interaction between HOXA13 and eIF4E in liver cancer cells and the deregulation of eIF4E mRNA and protein in cell cycle/nuclear pore HCC group phenotype and in T4 stage HCCs, respectively. Thus, transcriptional and post-transcriptional HOXA13 deregulation is involved in HCC possibly through the mRNA nuclear export of eIF4E-dependent transcripts. PMID- 21626506 TI - Risk of metachronous contralateral testicular germ cell tumors: a population based study of 7,102 Norwegian patients (1953-2007). AB - The purpose of the study was to identify overall incidence and risk of developing a metachronous contralateral testicular germ cell tumor (TGCT) and compare the risk for patients treated before and after 1980 (cisplatin became available for patients with metastatic TGCT). Our hypothesis was that the risk of metachronous TCGT would be reduced for patients with metastatic disease diagnosed after 1980. We included 7,102 men with unilateral TGCT, recorded in the Cancer Registry of Norway. Allowing for competing risk, cumulative incidence and adjusted hazard ratio (HR) were estimated for different subgroups, and the diagnostic periods 1953-1979 (I) and 1980-2007 (II). Relative risks were assessed by standardized incidence ratio (SIR). In Period I and Period II, 38 and 137 males, respectively, were diagnosed with metachronous contralateral TGCT. Corresponding 20-year cumulative incidences were 1.9% and 3.9%. In Period II, risk of a second TGCT was halved [HR = 0.5, 95% confidence interval (95% CI) = 0.33-0.77] for patients with metastatic compared to localized disease. For patients presenting with localized and metastatic disease, the SIRs for Period I were 14.6 (95% CI = 9.6-21.2) and 25.3 (95% CI = 12.1-46.5), respectively. In Period II, the corresponding numbers were 19.0 (95% CI = 15.6-22.9) and 9.8 (95% CI = 6.4-14.5). In conclusion, the risk of metachronous contralateral TGCT was halved for patients with metastatic compared to localized disease in Period II, whereas this protective effect of extent of disease lacked significance for Period I. These findings support our hypothesis that cisplatin-based chemotherapy reduced the risk of a second TGCT for patients with metastatic TGCT diagnosed after 1980. PMID- 21626507 TI - Fetal RHD genotype detection from circulating cell-free fetal DNA in maternal plasma in non-sensitized RhD negative women. AB - OBJECTIVE: To examine the performance of the SensiGene Fetal RHD Genotyping Laboratory Developed Test (RHD Genotyping LDT) using circulating cell-free fetal DNA (ccff DNA) extracted from maternal plasma. METHODS: ccff DNA was extracted from maternal blood from non-sensitized women with singleton pregnancies in two cohorts, one with a serotype reference (11-13 weeks' gestation) and one with the reference source (6-30 weeks' gestation). The presence of three RHD exon sequences (exons 4, 5, 7), the psi-pseudogene, three Y-chromosome sequences (SRY, DBY and TTTY2), and the X/Y-chromosome TGIF gene control were determined using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry-the RHD Genotyping LDT. RESULTS: The cohort with a serotype RhD reference showed correct classification in 201 of 207 patients, a test accuracy of 97.1%, with a sensitivity and specificity for prediction of RhD serotype of 97.2 and 96.8%, respectively. The cohort with a genotype RHD reference showed correct classification in 198 of 199 patients, indicating a test accuracy of 99.5% with a sensitivity and specificity for prediction of RHD genotype of 100.0 and 98.3%, respectively. CONCLUSION: Fetal RHD genotyping can accurately be determined using ccff DNA in the first and second trimesters of pregnancy. PMID- 21626508 TI - Mother-to-fetus transfer of antiviral drugs and the involvement of transporters at the placental barrier. AB - Administration of antiviral drugs to pregnant women with influenza, human immunodeficiency virus (HIV), and herpes simplex virus (HSV) infections is widely accepted as an effective treatment to safeguard the health of the mother and fetus. This review deals with the transfer of antiviral drugs to the fetus across the placental barrier, which is formed by an epithelial layer of syncytiotrophoblasts. First, the structure, function, and developmental change of the placenta and the placental barrier are briefly presented. We then review the transplacental permeability of antiviral drugs, such as oseltamivir for influenza, antiretrovirals (e.g., zidovudine, didanosine, and saquinavir) for HIV, and acyclovir for HSV, focusing on the involvement of ATP-binding cassette, organic anion/cation, and nucleoside transporters. The increasing evidence that is becoming available about transport mechanisms operating at the placental barrier is expected to be useful in the development of techniques to control fetal and placental transfer of antiviral drugs, and thereby to obtain maximum therapeutic benefit while minimizing potential fetal toxicity. PMID- 21626509 TI - Sugar ester J-1216 enhances percutaneous permeation of ionized lidocaine. AB - Percutaneous absorption enhancers affect not only the permeability of skin but also the thermodynamic properties of active ingredients in the vehicle. The present study examined the effect of J-1216, a sucrose laurate with hydrophilic lipophilic balance = 16, on the percutaneous permeation of lidocaine (LC) from this point of view. The percutaneous permeation of LC from aqueous vehicles (pH 6.0, 7.0, 8.0, and 10.0) with or without 1.5% J-1216 was examined with excised hairless mouse skin mounted on flow-through-type diffusion cell. The permeation of LC without J-1216 increased with an increase in the vehicle pH and could be basically explained by pH-partition theory. J-1216 increased the LC permeation at pH 6.0 and 7.0 but decreased it at pH 8.0 and 10.0. The interaction between LC and J-1216 was examined using an ultrafiltration technique. J-1216 micelles interacted predominantly with unionized LC. A theoretical calculation suggested that J-1216 enhances the permeability coefficient of ionized LC, whereas it has almost no effect on that of unionized free LC. J-1216 directly affects the skin to increase the permeation of ionized LC, whereas J-1216 micelles interact with unionized LC to decrease the permeation. The effect of J-1216 is therefore a function of vehicle pH and LC concentration. PMID- 21626510 TI - Gene cloning and expression of a novel hypoglycaemic peptide from Momordica charantia. AB - BACKGROUND: Momordica charantia (MC) is used in many Asian countries as a traditional functional food and medicine. Polypeptide-P, a 166 amino acid (AA) polypeptide isolated from MC seeds, has been reported to show hypoglycaemic effects in patients with type I or type II diabetes. The AA sequence of this peptide has been determined, but its gene sequence has yet to be published. RESULTS: In this study a gene-cloning strategy was employed to obtain the polypeptide-P gene sequence using degenerate reverse transcription polymer chain reaction and genome-walking methods. A complete 498 bp sequence encoding the polypeptide-P protein was cloned from MC seeds. Subsequent assays of the bioactivity of the expressed recombinant protein revealed that it had significant hypoglycaemic activity in alloxan-induced diabetic mice. This result suggests that recombinant polypeptide-P has hypoglycaemic effects. CONCLUSION: This is the first report of cloning and expression of the MC polypeptide-P gene. The cloned gene could be helpful for exploring the mechanisms of polypeptide-P gene expression and regulation in MC. Furthermore, this gene could be used as a potential tool both for screening MC varieties with high hypoglycaemically active substance content and for breeding new varieties of MC with high economic value, which could in turn be beneficial to farmers. PMID- 21626511 TI - Treatment outcome in the residually positive neck after definitive chemotherapy and irradiation. AB - OBJECTIVE: Determine prevalence of viable malignancy in patients undergoing neck dissection (ND) for residual neck disease following concomitant chemotherapy and irradiation (chemo/xrt) for upper aerodigestive squamous carcinoma. To determine survival in groups with a neck complete response to those who had residual disease requiring neck dissection. STUDY DESIGN: Retrospective chart review. METHODS: Retrospective chart review of 230 patients who underwent definitive chemo/xrt for primary squamous cell carcinoma cancer (SCCa) of the head and neck from 2005 to 2009 in one institution. RESULTS: Thirty-nine (17%) patients underwent ND for residual neck mass within 4 months posttreatment. Forty-nine percent (19/39) were pathologically positive for malignancy and 51% (20/39) were negative. The probability of a +ND based on original N-stage was not statistically significant (P = .368). Primary site did not yield significant probability of having +ND, except in the oral cavity (P = .02). Patients had similar overall 5-year survival, among those with a delayed complete response in the neck (66%), ND for residual disease (71%), or those with initial complete response (71%). Lower initial N-stage demonstrated improved survival in all outcome groups. Tonsil SCCa patients who underwent ND had improved survival compared to those with delayed complete response (87.5 vs. 75.8%), both of which had increased survival compared to initial complete responders (65%). CONCLUSIONS: This study supports the use of ND in the postchemo/xrt positive neck regardless of primary site or initial N-stage. ND in this setting conveys survival equal to patients with complete response in the neck after chemo/xrt. These survivorship implications of postchemo/xrt neck dissection extend to all sites, including tonsils. PMID- 21626512 TI - Value of contrast-enhanced sonography in the diagnosis of peripheral intrahepatic cholangiocarcinoma. AB - BACKGROUND: To illustrate contrast-enhanced harmonic ultrasonography (CEUS) findings of peripheral intrahepatic cholangiocarcinoma (PICC) and to assess the usefulness of CEUS in the diagnosis of this disease. METHODS: This study is a retrospective analysis of 46 patients (20 men, 26 women) with PICC who underwent both preoperative CEUS and pathologic examination. Sonographic examinations were performed with sonographic instruments equipped with low mechanical index contrast imaging software. The contrast agent SonoVue was used. The wash-in and wash-out features of the lesions were documented and the enhancement patterns were analyzed by visual inspection of the real-time cine loops. RESULTS: During the hepatic arterial phase of CEUS, the majority of PICC lesions (43/46, 93.5%) showed hypervascularity compared with the liver parenchyma, whereas three lesions (3/46, 6.5%) showed hypovascularity. In the portal phases, all lesions were either isoechoic or hypoechoic, while they were all hypoechoic in the late phase. Of all the hypervascular lesions, 48.8% (21/43) showed diffuse enhancement and 51.2% (22/43) showed rim-like or thick, rim-like enhancement. In 62.8% (27/43) of cases, progressive dendritic and centripetal fill-in of the contrast agent was observed. There were 83.7% (36/43) of hypervascular nodules showing heterogeneous enhancement during peak contrast enhancement of the lesions. CEUS provided clearer visualization of lesion margins than did conventional sonography. CONCLUSIONS: CEUS examination yields some specific findings that are useful in the diagnosis of PICC. PMID- 21626513 TI - Sonographic findings in a case of polyneuropathy associated with POEMS syndrome. AB - Polyneuropathy, organomegaly, endocrinopathy, M protein and skin changes (POEMS) syndrome is a rare multi-system disease. We describe the ultrasonography (US) and color Doppler ultrasonography (CDUS) findings of peripheral neuropathy in a patient with POEMS syndrome. In US examination, peripheral nerves were found to be diffusely thickned in both upper limbs. CDUS imaging showed arterial blood flow with low systolic peaks on the nerves. PMID- 21626514 TI - Sonographic visualization of a scapular body fracture: a case report. AB - We report the case of a scapular body fracture that was visualized with ultrasound and confirmed with three-dimensional CT. Ultrasound examination showed a comminuted fracture along the lateral border of the left scapula. We suggest scanning of the scapula, particularly the lateral scapula border, in patients with a history of shoulder trauma who continue to have symptoms in the shoulder girdle. PMID- 21626515 TI - Spontaneous thrombosis of uterine artery pseudoaneurysm: follow-up with Doppler ultrasonography and interventional management. PMID- 21626516 TI - Pararectal mass: an atypical location of splenosis. AB - Splenosis is the autotransplantation of splenic tissue resulting from the dissemination of cells from the pulp of the spleen after splenic injury or splenectomy. Implants can be found anywhere in the peritoneal cavity, especially on the serosal surfaces of small and large bowel, in the mesentery and diaphragm, implanted in visceral organs, within the thorax and brain, and in surgical scars and may vary in number, shape, and size. We described the sonographic, computed tomography and magnetic resonance imaging findings of pararectal splenosis in a 23-year-old man. The lesions appeared as multiple, well-circumscribed, small, round, homogenously solid masses of different sizes at the retrovesical and pelvic region detected during the imaging workup of Behcet disease. PMID- 21626517 TI - Research using blogs for data: public documents or private musings? AB - Nursing and other health sciences researchers increasingly find blogs to be valuable sources of information for investigating illness and other human health experiences. When researchers use blogs as their exclusive data source, they must discern the public/private aspects inherent in the nature of blogs in order to plan for appropriate protection of the bloggers' identities. Approaches to the protection of human subjects are poorly addressed when the human subject is a blogger and the blog is used as an exclusive source of data. Researchers may be assisted to protect human subjects via a decisional framework for assessing a blog author's intended position on the public/private continuum. PMID- 21626518 TI - In-line coupling SPE and CE for DNA preconcentration and separation. AB - An in-line SPE method coupled to CE was developed for the analysis of DNA. The amino silica monolith was prepared in situ by polymerization of tetraethoxysilane and N-(beta-aminoethyl)-gamma-aminopropyltriethoxysilane in ethanol aqueous solution at the inlet end of a 100 MUm id fused-silica capillary, and the remaining part of the capillary was used as separation channel. The procedure for this in-line SPE-CE method was constructed on the basis of investigation on operational conditions such as the introduction mode of sieving matrix, the composition of elution solvent and the elution time. Twenty millimolar ammonium hydroxide was demonstrated to be effective for DNA desorption from the monolith, and linear poly(N-isopropylacrylamide) was used as the separation matrix. The proposed method could achieve limits of detection of 0.065-0.123 ng/mL for six DNA fragments ranging 100-2000 bp. Compared with conventional CE, preconcentration factors of over 100 times were obtained. The applicability of the in-line SPE-CE method was further demonstrated by analyzing plasmid DNA from Escherichia coli crude lysate. PMID- 21626519 TI - Micropreparative isoelectric focusing protein separation in a suspended drop. AB - IEF protein binary separations were performed in a 12-MUL drop suspended between two palladium electrodes, using pH gradients created by electrolysis of simple buffers at low voltages (1.5-5 V). The dynamics of pH gradient formation and protein separation were investigated by computer simulation and experimentally via digital video microscope imaging in the presence and absence of pH indicator solution. Albumin, ferritin, myoglobin, and cytochrome c were used as model proteins. A drop containing 2.4 MUg of each protein was applied, electrophoresed, and allowed to evaporate until it splits to produce two fractions that were recovered by rinsing the electrodes with a few microliters of buffer. Analysis by gel electrophoresis revealed that anode and cathode fractions were depleted from high pI and low pI proteins, respectively, whereas proteins with intermediate pI values were recovered in both fractions. Comparable data were obtained with diluted bovine serum that was fortified with myoglobin and cytochrome c. PMID- 21626520 TI - Is metabolomics reachable? Different purification strategies of human colon cancer cells provide different CE-MS metabolite profiles. AB - In this work, four different metabolite purification approaches are investigated prior to metabolomics of human HT29 colon cancer cells. Namely, methanol deproteinization, ultrafiltration and two SPE methods using C18 and polymer-based cartridges were studied. The extracts were characterized via a metabolomic approach based on the application of CE TOF MS (CE-MS). CE-MS analysis time was less than 20 min per sample and allowed the simultaneous and reproducible analysis of more than 80 metabolites in a single run with a minimum consumption of sample and reagents. Metabolome analysis revealed in some cases important differences among the studied metabolite purification procedures. No significant differences were observed in the metabolite profile using C18 and polymer-based cartridges, or between ultrafiltration and methanol deproteinization. However, important differences were observed in the metabolomic profiles obtained from SPE and methanol deproteinization samples. These results demonstrate the crucial role of the metabolite purification strategy in metabolomics since it can bias (and in some cases mislead) the conclusions achieved by the metabolomic study. PMID- 21626521 TI - 2-DE and MALDI-TOF-MS analysis of therapeutic fusion protein abatacept. AB - 2-DE and MALDI-TOF MS are useful techniques for the quality evaluation of medicinal products derived from recombinant DNA technology. The principal objective of this study has been to evaluate the suitability of 2-DE in combination with MALDI-TOF MS for the quality study of the therapeutic recombinant protein, abatacept. 1-DE SDS-PAGE, under reducing and nonreducing conditions, and 2-DE analysis were used for the assessment of M(r) , pI, and enzymatic deglycosylation efficiency of abatacept. 2-DE allowed the assessment of product identity, purity, charge heterogeneity, isoform pattern, and post translational modifications. Furthermore, optimization of the deglycosylation procedure, charge heterogeneity, and sample preparation for the subsequent MALDI TOF MS analysis has been addressed. PMF analysis allowed rapid identity confirmation of abatacept. PMID- 21626522 TI - IRAP: An efficient retrotransposon-based electrophoretic technique for studying genetic variability among geographical isolates of Schistosoma japonicum. AB - In the present study, a inter-retrotransposon-amplified polymorphism (IRAP) technique, based on retrotransposons, was used to examine genetic variability among Schistosoma japonicum isolates from different provinces in mainland China. Of the 15 primers screened, 5 produced highly reproducible IRAP patterns. Using these primers, 54 discernible DNA fragments were generated with 40 (74.07%) being polymorphic, indicating considerable genetic variation among the examined S. japonicum isolates. The primer LTR-11 was found to be able to differentiate male and female parasites, producing one constant specific band for female S. japonicum isolates. The percentages of polymorphic bands (PPB) among all parasites, among isolates from mountainous provinces and among those from the lake/marshland areas were 74.07, 48.15, and 66.67%, respectively. UPGMA analysis revealed that the IRAP profiles could group S. japonicum isolates in mainland China into two clades (mountainous and lake/marshland types), and samples from the same geographical origins clustered together. These results demonstrated that the IRAP technique is suitable for studying genetic diversity and population structures, and also provides an effective technique for studying sex differentiation of S. japonicum. PMID- 21626523 TI - PCR-free digital minisatellite tandem repeat genotyping. AB - We demonstrated a proof-of-concept for novel minisatellite tandem repeat typing, called PCR-free digital VNTR (variable number tandem repeat) typing, which is composed of three steps: a ligation reaction instead of PCR thermal cycling, magnetic bead-based solid-phase capture for purification, and an elongated sample stacking microcapillary electrophoresis (MUCE) for sensitive digital coding of repeat number. We designed a 16-bp fluorescently labeled ligation probe which is complementary to a repeat unit of a biotinylated synthetic template mimicking the human D1S80 VNTR locus and is randomly hybridized with the minisatellite tandem repeats. A quick isothermal ligation reaction was followed to link the adjacent ligation probes on the DNA templates, and then the ligated products were purified by streptavidin-coated magnetic beads. After a denaturing step, a large amount of ligated products whose size difference was equivalent to the repeat unit were released and recovered. Through the elongated sample stacking MUCE separation on a microdevice, the fluorescence signal of the ligated products was generated in the electropherogram and the peak number was directly counted which was exactly matched with the repeat number of VNTR locus. We could successfully identify the minisatellite tandem repeat number with only 5 fmol of DNA template in 30 min. PMID- 21626525 TI - Limited use of interleukin 28B in the setting of response-guided treatment with detailed on-treatment virological monitoring. AB - A single-nucleotide polymorphism upstream of the interleukin-28B (IL28B) gene is associated with pegylated interferon-alfa-induced viral clearance in hepatitis C virus (HCV) genotype 1 patients. Using a well-characterized cohort of patients randomized to standard versus response-guided therapy, we studied whether the favorable CC type allows shortening of treatment duration. Association with viral kinetics, sustained viral response (SVR), and predictors of response were also analyzed. In the original study, 696 patients were randomized to either standard or variable therapy of 24, 48, or 72 weeks according to first undetectable HCV RNA. Association between IL28B determined by genotyping rs12979860 and end of treatment response and SVR by treatment arm was tested; baseline predictors of response were analyzed using multiple logistic regression. A total of 454 patients were evaluated. The frequency of IL28B type was CC = 29%, CT = 53%, TT = 18%. CC type was strongly associated with rapid virological response (RVR) as well as higher rates of week 8 and week 12 response. CC type was associated with SVR in both arms. In patients with RVR, SVR was high and IL28B type was not associated with SVR. In RVR patients, there was no significant difference in SVR or relapse rates after 24 or 48 weeks by IL28B type. Among non-RVR patients, CC type was associated with SVR at a higher rate than CT/TT, both in standard and variable analysis. However, when week 8 and week 12 responders were considered separately, IL28B type was no longer predictive of SVR. Few CC patients remained viremic beyond week 8 to allow the analysis of relationships between IL28B type and extended treatment. CONCLUSION: In HCV-1 patients, the favorable CC type strongly predicted higher rates of on-treatment virological milestones and SVR. However, achievement of on-treatment virological milestones was the critical factor in determining outcome. IL28B type appeared to have limited potential for response-guided treatment strategies. PMID- 21626524 TI - CXC chemokine signaling in the liver: impact on repair and regeneration. AB - The process of liver repair and regeneration following hepatic injury is complex and relies on a temporally coordinated integration of several key signaling pathways. Pathways activated by members of the CXC family of chemokines play important roles in the mechanisms of liver repair and regeneration through their effects on hepatocytes. However, little is known about the signaling pathways used by CXC chemokine receptors in hepatocytes. Here we review our current understanding of the pathways involved in both CXC chemokine receptor signaling in other cell types, most notably neutrophils, and similar pathways operant during hepatocyte proliferation/liver regeneration to formulate a basis for the function of CXC chemokine receptor signaling in hepatocytes. PMID- 21626526 TI - Compromising mitochondrial function with the antiretroviral drug efavirenz induces cell survival-promoting autophagy. AB - Hepatotoxicity is a very common side effect associated with the pharmacological treatment of human immunodeficiency virus (HIV) infection and its pathogenesis is poorly understood. Efavirenz (EFV) is the most widely used nonnucleoside reverse transcriptase inhibitor administered for the control of HIV and some of its toxic effects in hepatic cells have been recently shown to display features of mitochondrial dysfunction. Here we studied the activation of autophagy and, in particular, mitophagy, the main mitochondrial turnover mechanism, in human hepatic cells treated with clinically relevant concentrations of this drug. EFV treated cells had altered mitochondria, characterized by a relative increase in mitochondrial mass and defective morphology. This was followed by induction of autophagy as shown by the presence of autophagic vacuoles and the presence of the specific autophagic marker proteins microtubule-associated protein 1A/1B light chain 3 and Beclin-1. Importantly, whereas moderate levels of EFV activated autophagy, higher concentrations led to blockage in the autophagic flux, a condition that promotes "autophagic stress" and produces severe cellular damage. Finally, pharmacological inhibition of autophagy exacerbated the deleterious effect of EFV on cell survival/proliferation promoting apoptosis, which suggests that autophagy acts as an adaptive mechanism of cell survival. CONCLUSION: Clinical concentrations of EFV induce autophagy and, in particular, mitophagy in hepatic cells. Activation of this process promotes cell survival, but exceeding a certain threshold of mitochondrial dysfunction is associated with an autophagic overload or stress. This effect could be involved in the EFV-associated hepatotoxicity and may constitute a new mechanism implicated in the genesis of drug-induced liver damage. PMID- 21626527 TI - Rapid emergence of telaprevir resistant hepatitis C virus strain from wildtype clone in vivo. AB - Telaprevir is a potent inhibitor of hepatitis C virus (HCV) NS3-4A protease. However, the emergence of drug-resistant strains during therapy is a serious problem, and the susceptibility of resistant strains to interferon (IFN), as well as the details of the emergence of mutant strains in vivo, is not known. We previously established an infectious model of HCV using human hepatocyte chimeric mice. Using this system we investigated the biological properties and mode of emergence of mutants by ultra-deep sequencing technology. Chimeric mice were injected with serum samples obtained from a patient who had developed viral breakthrough during telaprevir monotherapy with strong selection for resistance mutations (A156F [92.6%]). Mice infected with the resistant strain (A156F [99.9%]) developed only low-level viremia and the virus was successfully eliminated with interferon therapy. As observed in patients, telaprevir monotherapy in viremic mice resulted in breakthrough, with selection for mutations that confer resistance to telaprevir (e.g., a high frequency of V36A [52.2%]). Mice were injected intrahepatically with HCV genotype 1b clone KT-9 with or without an introduced resistance mutation, A156S, in the NS3 region, and treated with telaprevir. Mice infected with the A156S strain developed lower level viremia compared to the wildtype strain but showed strong resistance to telaprevir treatment. Although mice injected with wildtype HCV showed a rapid decline in viremia at the beginning of therapy, a high frequency (11%) of telaprevir-resistant NS3 V36A variants emerged 2 weeks after the start of treatment. CONCLUSION: Using deep sequencing technology and a genetically engineered HCV infection system, we showed that the rapid emergence of telaprevir resistant HCV was induced by mutation from the wildtype strain of HCV in vivo. PMID- 21626528 TI - Hepatic triglyceride content in individuals with reduced intestinal cholesterol absorption due to variants in Nieman Pick C1-like 1. PMID- 21626531 TI - Acetaminophen overdose-induced liver injury in mice is mediated by peroxynitrite independently of the cyclophilin D-regulated permeability transition. AB - Acetaminophen (APAP) is safe at therapeutic dosage but can cause severe hepatotoxicity if used at overdose. The mechanisms of injury are not yet fully understood, but previous reports had suggested that the mitochondrial permeability transition (mPT) may be involved in triggering hepatocellular necrosis. We aimed at inhibiting mitochondrial cyclophilin D (CypD), a key regulator of the mPT, as a potential therapeutic target in APAP hepatotoxicity. Wildtype mice treated with a high dose of APAP (600 mg/kg, intraperitoneal) developed typical centrilobular necrosis, which could not, however, be prevented by cotreatment with the selective CypD inhibitor, Debio 025 (alisporivir, DEB025, a nonimmunosuppressive cyclosporin A analog). Similarly, genetic ablation of mitochondrial CypD in Ppif-null mice did not afford protection from APAP hepatotoxicity. To determine whether APAP-induced peroxynitrite stress might directly activate mitochondrial permeabilization, independently of the CypD regulated mPT, we coadministered the peroxynitrite decomposition catalyst Fe TMPyP (10 mg/kg, intraperitoneal, 90 minutes prior to APAP) to CypD-deficient mice. Liver injury was greatly attenuated by Fe-TMPyP pretreatment, and mitochondrial 3-nitrotyrosine adduct levels (peroxynitrite marker) were decreased. Acetaminophen treatment increased both the cytosolic and mitochondria associated P-JNK levels, but the c-jun-N-terminal kinase (JNK) signaling inhibitor SP600125 was hepatoprotective in wildtype mice only, indicating that the JNK pathway may not be critically involved in the absence of CypD. CONCLUSION: These data support the concept that an overdose of APAP results in liver injury that is refractory to pharmacological inhibition or genetic depletion of CypD and that peroxynitrite-mediated cell injury predominates in the absence of CypD. PMID- 21626532 TI - Random effects models for assessing diagnostic accuracy of traditional Chinese doctors in absence of a gold standard. AB - Two common problems in assessing the accuracy of traditional Chinese medicine (TCM) doctors in detecting a particular symptom are the unknown true symptom status and the ordinal-scale of the symptom status. Wang et al. (Biostatistics 2011; DOI: 10.1093/biostatistics/kxq075) proposed a nonparametric maximum likelihood method for estimating the accuracy of different TCM doctors without a gold standard when the true symptom status is measured on an ordinal-scale. A key assumption of their work is that the diagnosis results are independent conditional on the gold standard. This assumption can be violated in many practical situations.In this paper, we propose a random effects modeling approach that extends their method to incorporate dependence structure among different tests or doctors. The proposed method is illustrated on a real data set from TCM, which contains the diagnostic results from five doctors for the same patients regarding symptoms related to Chills disease. The same data set was analyzed by Wang et al. under the conditional independence assumption. In addition, we also discuss an ad hoc test for the model fitting and a likelihood ratio test on the random effects. PMID- 21626533 TI - Use of pretransformation to cope with extreme values in important candidate features. AB - Extreme values in predictors often strongly affect the results of statistical analyses in high-dimensional settings. Although they frequently occur with most high-throughput techniques, the problem is often ignored in the literature. We suggest to use a very simple transformation, proposed before in a different context by Royston and Sauerbrei, as an intermediary step between array preprocessing and high-level statistical analysis. This straightforward univariate transformation identifies extreme values in continuous features and can thus be used as a diagnostic tool for outliers. The use of the transformation and its effects is demonstrated for diverse univariate and multivariate statistical analyses using nine publicly available microarray data sets. PMID- 21626534 TI - Functional implications of variation in pectoral fin ray morphology between fishes with different patterns of pectoral fin use. AB - In this study, I compare the morphology from the pectoral fin rays from the benthic longhorn sculpin (Myoxocephalus octodecimspinosus) to those from a species that does not use its fins for substrate contact, the yellow perch (Perca flavescens). I use CT scanning technology to compare the shape and structure of the paired hemitrichia that make up the pectoral fin rays between these species. I found that the structure of hemitrichia of the fin rays in yellow perch is consistent with previous descriptions for pelagic fishes. They are almost completely segmented, have a crescent shape in cross section, and are branched distally. In contrast, longhorn sculpin hemitrichia exhibit morphological regionalization along the proximo-distal length of the ray. The most proximal 20 50% of the length of the hemitrichia is unsegmented and cylindrical in cross section. Distally, the fin rays of longhorn sculpin are segmented and crescent shaped but do not branch. I measured the second moment of area of the hemitrichia at distances of 10%, 30%, 50%, and 70% distance along the length of the fin rays. The cylindrical regions of the sculpin hemitrichia had a higher second moment of area than the crescent-shaped regions in either species. I hypothesize that that this regionalization of individual fin rays provides resistance to bending proximally and flexibility distally, features that may be useful during substrate contact. This combination of an elongate, unsegmented proximal region and segmented distal region in fin rays has not yet been described among extant ray finned fishes. However, this structure is reminiscent of that of the elongate cylindrical region found in the fossil sarcopterygian fish Eusthenopteron. PMID- 21626535 TI - Improved symptoms and lifestyle more than 20 years after untethering surgery for primary tethered cord syndrome. AB - AIMS: Evaluation of the long-term effectiveness of untethering surgery on social adaptations to urinary symptoms at the time of the survey. METHODS: Postoperative analysis of the current activities of daily life implemented as a postal questionnaire survey. Completed questionnaires were returned by 33 of 52 patients with primary tethered cord syndrome (p-TCS) who had undergone untethering surgery more than 20 years ago, including 10 patients with tight filum terminale (TFT) and 23 with cauda equina adhesion syndrome (CEAS). RESULTS: Long-term follow-up surveys in patients with p-TCS after untethering surgery indicated excellent effects on lower urinary tract symptoms in 9 of 10 patients with TFT, and the progression of symptoms tended to improve or halted in over half of the 23 patients with CEAS. However, one patient with TFT in whom new symptoms of voiding difficulty appeared immediately after surgery and 7 with CEAS showed deterioration of urinary and/or bowel symptoms. Eighteen patients complained of some symptoms caused by lower urinary tract dysfunction after untethering surgery in terms of difficulty in activities of daily life. With the exception of one female p-TCS patient with short stature and low body weight, all data of physical status, as well as educational level, employment, occupational category and marriage statistics were within the normal respective ranges taken from the Japanese census. CONCLUSIONS: Postoperative long-term follow-up survey in patients with p-TCS indicated significant improvement of urological symptoms. However, 55% of patients had some lower urinary tract dysfunction that affected their activities in daily life. PMID- 21626536 TI - Ethanol production by a new pentose-fermenting yeast strain, Scheffersomyces stipitis UFMG-IMH 43.2, isolated from the Brazilian forest. AB - The ability of a recently isolated Scheffersomyces stipitis strain (UFMG-IMH 43.2) to produce ethanol from xylose was evaluated. For the assays, a hemicellulosic hydrolysate produced by dilute acid hydrolysis of sugarcane bagasse was used as the fermentation medium. Initially, the necessity of adding nutrients (MgSO(4).7H(2)O, yeast extract and/or urea) to this medium was verified, and the yeast extract supplementation favoured ethanol production by the yeast. Then, in a second stage, assays under different initial xylose and cell concentrations, supplemented or not with yeast extract, were performed. All these three variables showed significant (p < 0.05) influence on ethanol production. The best results (ethanol yield and productivity of 0.19 g/g and 0.13 g/l/h, respectively) were obtained using the hydrolysate containing an initial xylose concentration of 30 g/l, supplemented with 5.0 g/l yeast extract and inoculated with an initial cell concentration of 2.0 g/l. S. stipitis UFMG-IMH 43.2 was demonstrated to be a yeast strain with potential for use in xylose conversion to ethanol. The establishment of the best fermentation conditions was also proved to be of great importance to increasing the product formation by this yeast strain. These findings open up new perspectives for the establishment of a feasible technology for ethanol production from hemicellulosic hydrolysates. PMID- 21626539 TI - Movement Disorders, the journal, and movement disorders, the field? PMID- 21626537 TI - Milestones in neuroimaging. AB - In the last 25 years there have been enormous advances in brain imaging. In addition to utility in diagnosis, these have led to novel insights into the pathogenesis of basal ganglia disease and the role of dopamine and the basal ganglia in normal health. The authors review highlights of this work, with a focus on advances in Parkinson's disease, the dystonias, Huntington's disease, and the role of dopamine in cognition and reward signaling. Emerging areas for future development include studies of functional connectivity, the analysis of default mode networks, studies of novel neurochemical pathways, methods to study disease pathogenesis, and the application of imaging techniques to investigate animal models of disease. PMID- 21626540 TI - The movement disorder society and movement disorders: a modern history. AB - The Movement Disorder Society (MDS) developed out of a merger with two short lived organizations, the Movement Disorder Society, primarily organized to develop a journal for the subspecialty, and the International Society of Motor Disturbances, primarily organized to develop international congresses. The formal merger of the Movement Disorder Society and the International Society of Motor Disturbances into the Movement Disorder Society took place at the 2nd International Congress of Movement Disorders in Munich, Germany, in June 1992. Whereas the journal, Movement Disorders, and the annual International Congress of Parkinson's Disease and Movement Disorders remain the anchors of the society, the goals now include the development of regional symposia, regional sections, Web based educational programs, and outreach efforts to include young investigators, wide international membership, and inclusion of non-neurologists, including basic scientists, neurosurgeons, and nonphysician health professionals. Movement Disorders has a continuingly growing subscribership and rising impact factor. PMID- 21626541 TI - Classification of movement disorders. AB - The classification of movement disorders has evolved. Even the terminology has shifted, from an anatomical one of extrapyramidal disorders to a phenomenological one of movement disorders. The history of how this shift came about is described. The history of both the definitions and the classifications of the various neurologic conditions is then reviewed. First is a review of movement disorders as a group; then, the evolving classifications for 3 of them--parkinsonism, dystonia, and tremor--are covered in detail. PMID- 21626542 TI - Milestones in clinical neurophysiology. AB - Over the last 25 years, clinical neurophysiology has made many advances in the understanding, diagnosis, and even treatment of different movement disorders. Transcranial magnetic stimulation has been the biggest technical advance. Progress in pathophysiology includes improved knowledge about bradykinesia in Parkinson's disease, loss of inhibition and increased plasticity in dystonia, abnormal startle in hyperekplexia, and various features of psychogenic movement disorders that can aid diagnosis. Studies have been done looking at the use of noninvasive brain stimulation for therapy, but effects are generally small. PMID- 21626543 TI - Milestones in magnetic resonance imaging and transcranial sonography of movement disorders. AB - Twenty-five years ago, when this journal was initiated, imaging of movement disorders was in its infancy. Since that time, magnetic resonance imaging has become a standard technique that is routinely performed in patients with movement disorders in order to exclude secondary causes and in some instances to provide specific information that aids in making the diagnosis of a neurodegenerative condition. Transcranial sonography is a more recent advance and is now widely employed to aid in the diagnosis of Parkinson's disease and possibly in detecting individuals in the premotor phases of the disease. Investigations are currently under way to evaluate the value of this technique in other movement disorders. PMID- 21626544 TI - A tale on animal models of Parkinson's disease. AB - Parkinson's disease is a neurodegenerative disorder whose cardinal manifestations are due primarily to a profound deficit in brain dopamine. Since the 1980s, several therapeutic strategies have been discovered to treat the symptoms of this neurological disorder, but as of yet, none halts or retards the neurodegenerative process. In an attempt to shed light on the neurobiology of Parkinson's disease, a number of experimental models have been developed, especially during the last 25 years. They come essentially in 3 flavors: pharmacological (eg, reserpine), toxic (eg, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine), and genetic (eg, transgenic synuclein mice). These models can also be recast as etiologic, pathogenic, and symptomatic/pathophysiologic, as each may contribute to our understanding of the cause, the mechanisms, and the treatment of Parkinson's disease. In this review, we will discuss the question of Parkinson's disease models, starting from the period when this journal was born to today. During this journey of 25 years, we will discuss both the significant contributions of the Parkinson's disease models and hurdles that remain to be overcome to one day cure this neurological disease. PMID- 21626545 TI - Milestones in movement disorders clinical trials: advances and landmark studies. AB - Over the past 25 years clinical trials testing in movement disorders has evolved in order to more effectively and efficiently analyze the safety and efficacy of new interventions. Studies today regularly incorporate methods to decrease placebo and bias effects and to ensure more rigorous statistical analyses. Newer, standardized, and validated rating scales such as the Unified Parkinson's Disease Rating Scale and the Unified Huntington's Disease Rating Scale are routinely employed in an effort to produce results that are comparable across different sites and studies. Several landmark studies in movement disorder research highlight these and other prominent procedural advances. The Deprenyl and Tocopherol Antioxidative Therapy of Parkinsonism trial pioneered the use of functional clinical end points, utilized a 2 * 2 factorial design to more efficiently analyze multiple interventions, and employed a washout design to assist in sorting putative neuroprotective from symptomatic effects. PRECEPT included neuroimaging as an outcome measure and highlighted the importance of futility studies in more efficiently directing resources. TEMPO and ADAGIO introduced the use of delayed-start (or 2-period) trials to try to identify disease-modifying interventions. NET-PD used futility studies to streamline the evaluation of potentially valuable treatments, followed by a large, long-term simple study design to assess the clinical significance of a new intervention. There have also been advances in clinical trials testing new surgical interventions, with the introduction of blinded outcome assessments and sham surgery control groups. Collectively, methodological advances in clinical trials have permitted the safety and efficacy of new interventions to be tested more efficiently and economically and with a higher level of certainty that the potential benefits and adverse effects of interventions recommended for general use are well understood. PMID- 21626546 TI - Milestones in Parkinson's disease--clinical and pathologic features. AB - The identification of the widespread deposition of fibrillized alpha-synuclein in Lewy bodies and Lewy neurites in the brains of patients with Parkinson's disease in 1997 has had a profound impact on how the disease is now conceptualized. The previous focus on the loss of the dopaminergic nigrostriatal system, the concept of subcortical dementia, and the idea that Parkinson's disease was dominated by motor impairment have all given way to research assessing more diverse brain regions, clinical symptoms, and phenotypes. It is now recognized that Parkinson's disease is more than just a loss of midbrain dopaminergic neurons in association with Lewy bodies. There are now several theories on how the disease develops and progresses currently being validated in a variety of studies, although many of these theories have yet to incorporate the phenotypic clinical and pathological changes associated with age. A particularly exciting new area of research involves the cell-to-cell transmission of pathogenic proteins. The recent consensus definition of Parkinson's disease dementia will allow its pathologic substrates to be determined. These advances have progressed to a stage where the preclinical stages of Parkinson's disease and its specific signs and symptoms are being predicted and tested clinically. Such strategies herald a future wave of preventive strategies for Parkinson's disease and its clinical symptoms. PMID- 21626547 TI - Parkinson's disease: the quintessential neuropsychiatric disorder. AB - Although diagnosed by characteristic motor features, Parkinson's disease may be preceded, and is frequently accompanied by, a wide range of cognitive and neuropsychiatric features. In addition to the most commonly studied disorders of dementia, depression, and psychosis, other relatively common and clinically significant psychiatric complications include impulse control disorders, anxiety symptoms, disorders of sleep and wakefulness, and apathy. These problems may be underrecognized and are frequently undertreated. The emergent focus on nonmotor aspects of Parkinson's disease over the past quarter of a century is highlighted by a nonlinear increase in the number of articles published devoted to this topic. Although the development of newer antidepressants, atypical antipsychotics, and cholinesterase inhibitors in recent years has had a positive benefit on the management of these troublesome and distressing symptoms, responses are frequently suboptimal, and this remains an area of major unmet therapeutic need. PMID- 21626549 TI - Milestones in PD genetics. AB - Over the last 25 years, genetic findings have profoundly changed our views on the etiology of Parkinson's disease. Linkage studies and positional cloning strategies have identified mutations in a number of genes that cause several monogenic autosomal-dominant or autosomal-recessive forms of the disorder. Although most of these Mendelian forms of Parkinson's disease are rare, whole genome association studies have more recently provided convincing evidence that low-penetrance variants in at least some of these, but also in several other genes, play a direct role in the etiology of the common sporadic disease as well. In addition, rare variants with intermediate-effect strengths in genes such as Gaucher's disease-associated glucocerebrosidase A have been discovered as important risk factors. "Next-generation" sequencing technologies are expected by some to identify many more of these variants. Thus, an increasingly complex network of genes contributing in different ways to disease risk and progression is emerging. These findings may provide the "genetic entry points" to identify molecular targets and readouts necessary to design rational disease-modifying treatments. PMID- 21626548 TI - Milestones in research on the pathophysiology of Parkinson's disease. AB - Progress in our understanding of the mechanisms underlying the cardinal motor abnormalities of Parkinson's disease (PD), in particular akinesia and bradykinesia and their treatment, has been remarkable. Notable accomplishments include insights into the functional organization of the basal ganglia and their place in the motor system as components of a family of parallel cortico subcortical circuits that subserve motor and nonmotor functions and the development of models of the intrinsic organization of the basal ganglia, including delineation of the so-called direct, indirect, and hyperdirect pathways. Studies in primate models of PD have provided insight into the alterations of neuronal activity that are responsible for the motor features of PD, revealing both altered tonic levels of discharge and significant disturbances of the patterns of discharge throughout the motor circuitry and have led to the formulation of circuit models of PD, providing testable hypotheses for research and stimulating the development of new therapies. Most importantly, the discovery that lesions of the subthalamic nucleus, a key node of the indirect pathway, abolish the cardinal features of PD contributed to the renaissance in the use of surgical approaches to treating patients with PD, including ablation and deep brain stimulation. PMID- 21626550 TI - Etiology and pathogenesis of Parkinson's disease. AB - The past 25 years have seen a major expansion of knowledge concerning the cause of Parkinson's disease provided by an understanding of environmental and genetic factors that underlie the loss of nigral dopaminergic neurons. Based on the actions of toxins, postmortem investigations, and gene defects responsible for familial Parkinson's disease, there is now a general consensus about the mechanisms of cell death that contribute to neuronal loss in Parkinson's disease. Mitochondrial dysfunction, oxidative stress, altered protein handling, and inflammatory change are considered to lead to cell dysfunction and death by apoptosis or autophagy. Ageing is the single most important risk factor for Parkinson's disease, and the biochemical changes that are a consequence of aging amplify these abnormalities in Parkinson's disease brain. What remains to be determined is the combination and sequence of events leading to cell death and whether this is identical in all brain regions where pathology occurs and in all individuals with Parkinson's disease. Focusing on those events that characterize Parkinson's disease, namely, mitochondrial dysfunction and Lewy body formation, may be the key to further advancing the understanding of pathogenesis and to taking these mechanisms forward as a means of defining targets for neuroprotection. PMID- 21626551 TI - Parkinson's disease, proteins, and prions: milestones. AB - Parkinson's disease (PD) is characterized by protein accumulation in the form of Lewy bodies and neurites. It is thus reasonable to consider that alterations in protein handling in the form of increased production, impaired clearance, or both might be central to the etiopathogenesis of the disease. Increasing genetic, laboratory and pathologic evidence has accumulated over the past 25 years supporting this hypothesis. A vicious cycle could develop in which increased protein accumulation from any cause could lead to interference with lysosomal and proteasomal clearance mechanisms causing further protein accumulation. Eventually, protein accumulation could overwhelm the cell's defenses and lead to the formation of toxic oligomers and amyloid-based inclusions such as Lewy bodies, disruption of critical cell processes, and ultimately neurodegeneration. More recent findings of Lewy pathology in implanted embryonic dopamine neurons in PD patients raises the intriguing possibility that PD might be a prion disorder. These concepts suggests new targets and novel candidate therapies that might be neuroprotective for PD. PMID- 21626552 TI - Milestones in Parkinson's disease therapeutics. AB - In the mid-1980s, the treatment of Parkinson's disease was quite exclusively centered on dopatherapy and was focusing on dopamine systems and motor symptoms. A few dopamine agonists and a monoamine oxidase B inhibitor (selegiline) were used as adjuncts in advanced Parkinson's disease. In the early 2010s, levodopa remains the gold standard. New insights into the organization of the basal ganglia paved the way for deep brain stimulation, especially of the subthalamic nucleus, providing spectacular improvement of drug-refractory levodopa-induced motor complications. Novel dopamine agonists (pramipexole, ropinirole, rotigotine), catecholmethyltransferase inhibitors (entacapone), and monoamine oxidase B inhibitors (rasagiline) have also been developed to provide more continuous oral delivery of dopaminergic stimulation in order to improve motor outcomes. Using dopamine agonists early, before levodopa, proved to delay the onset of dyskinesia, although this is achieved at the price of potentially disabling daytime somnolence or impulse control disorders. The demonstration of an antidyskinetic effect of the glutamate antagonist amantadine opened the door for novel nondopaminergic approaches of Parkinson's disease therapy. More recently, nonmotor symptoms (depression, dementia, and psychosis) have been the focus of the first randomized controlled trials in this field. Despite therapeutic advances, Parkinson's disease continues to be a relentlessly progressive disorder leading to severe disability. Neuroprotective interventions able to modify the progression of Parkinson's disease have stood out as a failed therapeutic goal over the last 2 decades, despite potentially encouraging results with compounds like rasagiline. Newer molecular targets, new animal models, novel clinical trial designs, and biomarkers to assess disease modification have created hope for future therapeutic interventions. PMID- 21626553 TI - Milestones in atypical and secondary Parkinsonisms. AB - During the last decades, atypical parkinsonian disorders such as multiple system atrophy, dementia with Lewy bodies, progressive supranuclear palsy, and corticobasal degeneration along with secondary parkinsonian disorders have been increasingly recognized as important causes of parkinsonism. Although treatment options are largely limited to date, remarkable progress has occurred through advances in the fields of molecular biology and diagnostic neuroimaging, resulting in intense preclinical drug discovery programs. Early-investigation assisted clinical diagnosis has become more crucial than ever because disease modifying therapies will hopefully become available within this decade. PMID- 21626554 TI - Milestones in tremor research. AB - Tremor is one of the most frequent movement disorders and covers a wide spectrum of entities summarized in the 1998 consensus statement of the Movement Disorder Society. Essential tremor and Parkinson tremor are most common and are also the most thoroughly studied. Major progress has occurred in the clinical semiology, neuroimaging, epidemiology, and pathophysiology of tremors. Pathology and genetic research are rapidly growing fields of study. Recently described tremor entities include orthostatic tremor, dystonic tremor, cortical tremor, and thalamic tremor. Treatment research methodology has improved substantially, but few double blind controlled trials have been published. Deep brain stimulation is the most effective treatment for most tremors but is reserved for advanced cases. PMID- 21626555 TI - Milestones in dystonia. AB - The last 25 years have seen remarkable advances in our understanding of the genetic etiologies of dystonia, new approaches into dissecting underlying pathophysiology, and independent progress in identifying effective treatments. In this review we highlight some of these advances, especially the genetic findings that have taken us from phenomenological to molecular-based diagnoses. Twenty DYT loci have been designated and 10 genes identified, all based on linkage analyses in families. Hand in hand with these genetic findings, neurophysiological and imaging techniques have been employed that have helped illuminate the similarities and differences among the various etiological dystonia subtypes. This knowledge is just beginning to yield new approaches to treatment including those based on DYT1 animal models. Despite the lag in identifying genetically based therapies, effective treatments, including impressive benefits from deep brain stimulation and botulinum toxin chemodenervation, have marked the last 25 years. The challenge ahead includes continued advancement into understanding dystonia's many underlying causes and associated pathology and using this knowledge to advance treatment including preventing genetic disease expression. PMID- 21626556 TI - Milestones in huntington disease. AB - There have been extraordinary advances in our knowledge of the underlying gene, the protein it encodes, various models of disease, and potential targets for effective therapies for Huntington disease. Huntington disease research has increased exponentially in the past 25 years, and we now understand many of the molecular mechanisms underlying the disease. Still, more work needs to be done before we have a full understanding of the pathophysiology of the disease. Clinical research on biomarkers and clinical trials on potential neuroprotective agents are underway. Here we review our progress in these areas over the last 25 years and speculate on what the next 25 years may hold. PMID- 21626558 TI - Milestones in myoclonus. AB - This review examines some of the advances in understanding myoclonus over the last 25 years. The classification of myoclonus into cortical, brainstem, and spinal forms has been consolidated, each with distinctive clinical characteristics and physiological mechanisms. New genetic causes of myoclonus have been identified, and the molecular basis of several of these conditions has been discovered. It is increasingly apparent that disease of the cerebellum is particularly important in the genesis of cortical reflex myoclonus. However, the precise mechanism and origin of myoclonus in many situations remain uncertain. Effective treatment of myoclonus remains limited, and the challenge lies ahead to develop more therapeutic options. PMID- 21626557 TI - Milestones in ataxia. AB - The past 25 years have seen enormous progress in the deciphering of the genetic and molecular basis of ataxias, resulting in improved understanding of their pathogenesis. The most significant milestones during this period were the cloning of the genes associated with the common spinocerebellar ataxias, ataxia telangiectasia, and Friedreich ataxia. To date, the causative mutations of more than 30 spinocerebellar ataxias and 20 recessive ataxias have been identified. In addition, there are numerous acquired ataxias with defined molecular causes, so that the entire number of distinct ataxia disorders exceeds 50 and possibly approaches 100. Despite this enormous heterogeneity, a few recurrent pathophysiological themes stand out. These include protein aggregation, failure of protein homeostasis, perturbations in ion channel function, defects in DNA repair, and mitochondrial dysfunction. The clinical phenotypes of the most common ataxia disorders have been firmly established, and their natural history is being studied in ongoing large observational trials. Effective therapies for ataxias are still lacking. However, novel drug targets are under investigation, and it is expected that there will be an increasing number of therapeutic trials in ataxia. PMID- 21626559 TI - Paroxysmal dyskinesias. AB - Paroxysmal movement disorders are a relatively rare and heterogenous group of conditions manifesting as episodic dyskinesia lasting a brief duration. Three forms are clearly recognized, namely, paroxysmal kinesigenic (PKD), nonkinisegenic (PNKD), and exercise induced (PED). There have been major advances in the understanding of the pathophysiological mechanisms and the genetics of these disorders, leading to better clinical definitions based on genotype phenotype correlations in the familial idiopathic forms. PKD is genetically heterogenous, but there is linkage to chromosome 16 in a number of families. PNKD is due to mutations of the MR-1 gene. PED is genetically heterogenous, but a number of familial and sporadic cases may be due to GLUT-1 gene mutations. The GLUT1 gene-related form of PED may respond to a ketogenic diet. Potassium and calcium channel mutations underlie the 2 main forms of episodic ataxia (EA1 and EA2), whereas benign torticollis of infancy may also be a calcium channel disorder. PMID- 21626560 TI - Milestones in gait, balance, and falling. AB - Gait, balance, and falls have become increasingly common topics of published articles in the Movement Disorders journal since its launch in 1986. This growth represents an increasing awareness of the importance of mobility to patients' quality of life. New methods have become available that allow for accurate measurement of many aspects for gait and balance. This has led to new concepts of understanding gait and balance disorders. Neuroimaging has begun to reveal the neural circuitry underlying gait and balance. The physiology and pathophysiology of balance and gait are beginning to tease out the many processes involved in mobility and how they may be disrupted by disease processes. With these advances, the old therapeutic nihilism that characterized the clinician's approach to falls and gait disorders is disappearing, as innovative physiotherapy, exercise, drugs, and deep brain stimulation are being employed for gait and balance disorders. PMID- 21626561 TI - Psychogenic movement disorders: past developments, current status, and future directions. AB - As the field of movement disorders has developed and matured over the past 25 years, psychogenic movement disorders have become increasingly recognized in subspecialty clinics. The diagnosis can be challenging and should be based on positive features rather than a purely exclusionary approach. The clinical phenotype can be quite varied, although certain categories of abnormal movement are more common than others. Electrophysiological studies may be particularly useful in establishing the diagnosis, especially with respect to tremor and myoclonus, and an argument can be made for adding a "laboratory-supported definite" category to earlier classification schemes. The diagnosis of psychogenic dystonia remains a major challenge, although there are some recent promising developments with respect to the evaluation of cortical plasticity that require further study. The pathogenesis of psychogenic movement disorders is poorly understood; insights may be provided from the study of other neurological conversion disorders such as psychogenic hemiparesis. Psychogenic movement disorders typically result in considerable disability and negatively impact quality of life to the same or greater extent than do many organic movement disorders. Treatment is extremely challenging, and many patients experience chronic disability despite various therapeutic interventions. Given the personal and societal impact of these problems, further advances in our understanding of their pathogenesis and the subsequent development of effective therapies are sorely needed. PMID- 21626562 TI - Lack of association of the UCHL-1 gene with Parkinson's disease in a Greek cohort: a haplotype-tagging approach. PMID- 21626563 TI - Ability to cycle despite severe freezing of gait in atypical parkinsonism in Fahr's syndrome. PMID- 21626564 TI - Patients' perceptions of life shift after deep brain stimulation for primary dystonia--A qualitative study. AB - Studies of deep brain stimulation for dystonia have shown significant motor improvement. However, patients' perceptions of surgery and its effects have been less studied. We aimed to explore perceptions of changes in life in patients with primary dystonia after deep brain stimulation. Thirteen patients underwent thematic interviews 8-60 months after pallidal deep brain stimulation. Interviews were transcribed verbatim and analyzed with grounded theory. Patients described a profound impact of dystonia on daily life. After surgery, physical changes with a more upright posture and fewer spasms translated into an easier, more satisfying life with greater confidence. Notwithstanding this positive outcome, the transition from a limited life before surgery to opportunities for a better life exhibited obstacles: The "new life" after deep brain stimulation was stressful, including concern about being dependent on the stimulator as well as having to deal with interfering side effects from deep brain stimulation. The whole coping process meant that patients had to quickly shift focus from struggling to adapt to a slowly progressive disorder to adjustment to a life with possibilities, but also with new challenges. In this demanding transition process, patients wished to be offered better professional guidance and support. Even though deep brain stimulation provides people with primary dystonia with a potential for better mobility and more confidence, patients experienced new challenges and expressed the need for support and counseling after surgery. Grounded theory is a useful method to highlight patients' own experience and contributes to a deeper understanding of the impact of deep brain stimulation on patients with dystonia. PMID- 21626565 TI - Brief cognitive tests in the screening of dementia in Parkinson's disease. PMID- 21626567 TI - Comparison of cerebellar ataxias: A three-year prospective longitudinal assessment. AB - We quantitatively investigated the clinical severity and progression of diseases with ataxia, as measured with the Scale for the Assessment and Rating of Ataxia, and examined the potential application of the Scale for the Assessment and Rating of Ataxia for future therapeutic trials. Severity of ataxia was assessed in 238 patients with spinocerebellar ataxia type 2, spinocerebellar ataxia type 3, spinocerebellar ataxia type 6, spinocerebellar ataxia type 17, multiple system atrophy-cerebellar variant, or Gerstman-Straussler-Scheinker disease. Among them, 119 (50%) were longitudinally examined three to seven times, in a period of 8 to 38 months, resulting in a total set of 535 assessments. The differences between spinocerebellar ataxia and multiple system atrophy-cerebellar variant were ascertained cross-sectionally and longitudinally. Gerstman-Straussler-Scheinker disease had the fastest progression, followed by multiple system atrophy cerebellar variant, spinocerebellar ataxia type 17, spinocerebellar ataxia type 3, spinocerebellar ataxia type 2, and spinocerebellar ataxia type 6. Patients with multiple system atrophy-cerebellar variant had a faster progression in gait, sitting, speech, and total score than patients with spinocerebellar ataxias. For a randomized, case-control trial, a sample size of 47 for spinocerebellar ataxia and 85 for multiple system atrophy-cerebellar variant in the treatment or placebo arms would have a sufficient statistical power to demonstrate the efficacy of a new therapy that would retard ataxia progression by 1 point per year as measured by the Scale for the Assessment and Rating of Ataxia. The results will have a significant impact on the planning and implementation of future therapeutic trials of spinocerebellar ataxia and multiple system atrophy-cerebellar variant. PMID- 21626568 TI - The accuracy of family physicians' dementia diagnoses at different stages of dementia: a systematic review. AB - OBJECTIVE: Optimising care for dementia patients and their informal carers is imperative in light of the impending dementia epidemic. An important aspect of care is accurate recognition and diagnosis of dementia. The aim of this review was to estimate family physicians' diagnostic accuracy at the different stages of dementia. METHODS: Pubmed, Embase, CINAHL, PsycINFO and the Cochrane Library were searched for articles comparing family physicians' 'dementia' and 'cognitive impairment' diagnoses in the primary care setting to reference standard dementia diagnoses. RESULTS: Data from six cross-sectional studies of moderate to reasonable methodological quality were extracted for the analysis. One study considered the accuracy of family physicians' recollected diagnoses, and three studies focussed on documented diagnoses. In these four studies, the sensitivity of family physicians' combined diagnostic categories of 'cognitive impairment' together with 'dementia' was 0.48-0.67 for mild dementia and 0.76-0.85 for moderate to severe dementia. The sensitivity of their diagnostic category 'dementia' alone was 0.14-0.33 for mild and 0.28-0.61 for moderate to severe dementia. Specificity was excellent for all severity stages in both comparisons. Three studies examined the accuracy of family physicians' judgement of cognition during consultation. Compared with the studies on recollection and documentation, these studies reported higher sensitivity and lower specificity. CONCLUSION: Many individuals with dementia are not recognised or not diagnosed as such; particularly mild dementia is under-diagnosed. Collaboration within primary care and education focussing both on knowledge and attitude are recommended to improve the accuracy of family physicians' dementia diagnosis. PMID- 21626569 TI - Subtle cognitive impairment in elders with Mini-Mental State Examination scores within the 'normal' range. AB - OBJECTIVE: The Mini-Mental State Examination (MMSE) is commonly used as a screening test for dementia, yet MMSE scores above the cut-off for dementia (24 30) are widely thought to have limited utility, particularly in older persons. The study investigates whether scores within this range can be indicative of pre symptomatic levels of cognitive impairment. METHODS: Ninety-six community dwelling older persons aged 62-89 years (mean = 75.2 years), who had obtained MMSE scores between 25 and 30, were tested on the computer-based Subtle Cognitive Impairment Test (SCIT). RESULTS: Compared with individuals who obtained a perfect score of 30 on the MMSE, individuals with scores of 28-29 made more errors on the SCIT, whereas those with scores of 25-27 on the MMSE made the most errors on the SCIT (F(2,94) = 9.84, p < 0.01). Individuals who made errors in the language (r(94) = -0.47, p < 0.01), attention (r(94) = 0.24, p < 0.05) and visual construction (r(94) = -0.27, p < 0.01) subtests of the MMSE were more likely to display impaired SCIT performance. CONCLUSIONS: The pattern of performance on the SCIT varied in a systematic way, depending on the MMSE subtest in which the errors were made, raising the possibility that there may be different subtypes of subtle cognitive impairment within the ostensibly normal population of older persons. PMID- 21626570 TI - Association of grip strength with dementia in a Korean older population. AB - BACKGROUND: Several studies have found that low grip strength is associated with dementia in Western populations. However, there have been few studies in Asian populations. This cross-sectional study aimed to examine whether grip strength is independently associated with dementia after controlling for other related factors in a Korean older population. METHODS: One thousand thirty-eight community-dwelling older people aged 65 or over within a defined geographic area were screened for dementia. Data on grip strength, sit-to-stand score, body mass index (BMI), socio-demographic factors (age, gender, marital status, education), and medical conditions (diabetes, hypertension, heart disease, stroke) were collected. Dementia was diagnosed with the 10/66 dementia diagnostic algorithm. RESULTS: Eleven percent of the participants were found to have dementia. After adjustment for other factors, grip strength (per 8-kg decrease) was independently associated with dementia (odds ratio, 1.59; 95% confidence interval, 1.19-2.14). Adjusted grip strength by BMI interaction terms showed marginal significance (p = 0.098); that is, older people with both reduced grip strength and low BMI were likely to be at higher risk for dementia. CONCLUSIONS: Reduced grip strength was independently associated with dementia in a Korean older population. This association was partially influenced by BMI. PMID- 21626571 TI - Expression of plasminogen activator inhibitor-1 by olfactory ensheathing glia promotes axonal regeneration. AB - Olfactory ensheathing glia (OEG) cells are known to facilitate repair following axotomy of adult neurons, although the molecular mechanisms involved are not fully understood. We previously identified plasminogen activator inhibitor-1 (PAI 1), proteinase-activated receptor-1 (PAR-1), and thrombomodulin (TM) as candidates to regulate rat OEG-dependent axonal regeneration. In this study, we have validated the involvement of these proteins in promoting axonal regeneration by immortalized human OEGs. We studied the effect of silencing these proteins in OEGs on their capacity to promote the regeneration of severed adult retinal ganglion cells (RGCs) axons. Our results support the role of glial PAI-1 as a downstream effector of PAR-1 in promoting axon regeneration. In contrast, we found that TM inhibits OEG induced-axonal regeneration. We also assessed the signaling pathways downstream of PAR-1 that might modulate PAI-1 expression, observing that specifically inhibiting Galpha(i), Rho kinase, or PLC and PKC downregulated the expression of PAI-1 in OEGs, with a concomitant reduction in OEG-dependent axon regeneration in adult RGCs. Our findings support an important role for the thrombin system in regulating adult axonal regeneration by OEGs. PMID- 21626572 TI - Identifying patterns of symptom change during a randomized controlled trial of cognitive processing therapy for military-related posttraumatic stress disorder. AB - Cognitive processing therapy (CPT) for posttraumatic stress disorder (PTSD) has been shown to reduce symptoms of PTSD in a veteran population. This study explored patterns of self-reported symptom change during CPT. Veterans (N = 60) with PTSD were randomized to receive CPT immediately or after 10 weeks. We hypothesized that those treated immediately would evidence initial symptom stability followed by decline compared with those who waited, whose PTSD symptoms would remain stable. The best model fit based on deviance statistics and Bayesian information criteria comparisons was one in which participants treated immediately showed more rapid initial decline followed by a slower rate of PTSD symptom improvement relative to those who waited, who showed a stable level of symptomatology. Findings suggest that CPT produces quick and maintained improvements in veterans. The effect sizes for change between those who received CPT immediately and those who waited were approximately medium sized. Implications of findings are discussed. PMID- 21626573 TI - Exploring the efficacy of a residential treatment program incorporating cognitive processing therapy-cognitive for veterans with PTSD and traumatic brain injury. AB - As the numbers of military personnel participating in the wars in Afghanistan and Iraq continue to grow, the percentage of individuals who return with both a traumatic brain injury (TBI) and posttraumatic stress disorder (PTSD) also increases. Although there appears to be significant overlap in the symptoms resulting from PTSD and TBI, the best course of treatment remains an area of controversy. The authors present initial findings from a Veterans Administration residential program for comorbid PTSD and TBI. Forty-two participants completed a program comprising psychoeducational groups and cognitive skill building that was augmented with a modification of standard cognitive processing therapy. The results suggest that residential programs that incorporate this form of cognitive therapy can anticipate meaningful participation from patients, and that it may be an effective approach to treat PTSD in individuals with a history of TBI. PMID- 21626574 TI - Risky business: trauma exposure and rate of posttraumatic stress disorder in African American children and adolescents. AB - Demographics, parental risk factors, and experiencing interpersonal trauma (domestic violence, community violence, and physical and sexual abuse) are related to childhood posttraumatic stress disorder (PTSD). Little is known about these factors and the risk of PTSD in African American children. This study examined associations between PTSD symptoms and gender, age, parent mental illness, parent substance abuse, and interpersonal trauma in African American children. Participants were 257 children and adolescents, ages 8-17 years (M = 11.7, SD = 2.5), who received outpatient mental health treatment. Being female and witnessing domestic violence was associated with more PTSD symptoms. Exposure to community violence and physical abuse increased the odds of clinically significant PTSD symptomatology by more than 2 times. The rate of PTSD (16%) was lower in the current study than in other same-age study populations (25%-40%). Risk factors and identification strategies for PTSD are discussed. PMID- 21626576 TI - Transparent conductive graphene films synthesized by ambient pressure chemical vapor deposition used as the front electrode of CdTe solar cells. PMID- 21626577 TI - Dynamic topographical control of mesenchymal stem cells by culture on responsive poly(epsilon-caprolactone) surfaces. PMID- 21626578 TI - Highly efficient activation of molecular oxygen with nanoporous metalloporphyrin frameworks in heterogeneous systems. PMID- 21626579 TI - Facile fabrication of hollow mesoporous silica nanospheres for superhydrophilic and visible/near-IR antireflection coatings. AB - A series of hierarchically mesostructured silica nanoparticles (MSNs) less than 100 nm in size were fabricated by means of a one-step synthesis using dodecanethiol (C(12)-SH) and cetyltrimethylammonium bromide (CTAB) as the dual template, and trimethylbenzene (TMB) as the swelling agent. Silica nanoparticles with varied morphologies and structures, including mesoporous silica nanoparticles with tunable pore size, mesoporous silica nanoparticles with a thin solid shell, hollow mesoporous silica nanoparticles with tunable cavity size, and hollow mesoporous silica nanoparticles with a thin solid shell, were obtained by regulating the TMB/CTAB molar ratio and the stirring rate with the assistance of C(12)-SH. Silica particulate coatings were successfully fabricated by using MSNs with varied morphologies and structures as building block through layer-by-layer dip-coating on glass substrates. The thickness and roughness of the silica particulate coatings could be tailored by regulating the deposition cycles of nanoparticles. The silica particulate coatings composed of hollow mesoporous silica nanoparticles with a thin shell (S2) increased the maximum transmittance of slide glass from 90 to 96%, whereas they reduced its minimum reflection from 8 to 2% at the optimized wavelength region that could be adjusted from visible to near-IR with a growing number of deposition cycles. The coatings also exhibited excellent superhydrophilic and antifogging properties. These mesostructured silica nanoparticles are also expected to serve as ideal scaffolds for biological, medical, and catalytic applications. PMID- 21626580 TI - Spectroelectrochemical evidence for the nitrosyl redox siblings NO+, NO*, and NO- coordinated to a strongly electron-accepting Fe(II) porphyrin: DFT calculations suggest the presence of high-spin states after reduction of the Fe(II)-NO- complex. AB - Experimental and computational results for the electron-deficient porphyrin complex [Fe(NO)(TFPPBr(8))] (1; TFPPBr(8)=2,3,7,8,12,13,17,18-octabromo 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin) are reported with respect to its electron-transfer behavior. Complex 1 undergoes three one-electron processes: two reversible reductions and one irreversible oxidation. Spectroelectrochemical measurements (IR and UV/Vis/NIR spectroscopy) of (14)NO- and (15)NO-containing material indicate that the first reduction to 1(-) occurs largely on the NO ligand to produce nitroxyl anion (NO(-)) character, as evident from the considerable change in nu(NO) from 1715 to around 1550 cm(-1). The second reduction to 1(2)(-) does not result in a further shift of nu(NO) to lower frequencies, but to a surprising high-energy shift to 1590 cm(-1). This and the notable changes of the characteristic porphyrin vibrations as well as significant changes of the UV/Vis absorptions indicate a porphyrin-centered process; DFT calculations predict the shift of nu(NO) to higher frequencies for the intermediate- and high-spin states of 1(2-). The oxidation of 1 is irreversible on the voltammetry timescale, but chemically reversible in spectroelectrochemical experiments, suggesting that the cationic form dissociates to the corresponding ferric porphyrin and NO. DFT calculations support the interpretation of the experimental results. PMID- 21626581 TI - Understanding the acid-base properties of adenosine: the intrinsic basicities of N1, N3 and N7. AB - Adenosine (Ado) can accept three protons, at N1, N3, and N7, to give H(3) (Ado)(3+) , and thus has three macro acidity constants. Unfortunately, these constants do not reflect the real basicity of the N sites due to internal repulsions, for example, between (N1)H(+) and (N7)H(+). However, these macroconstants are still needed for the evaluations and the first two are taken from our own earlier work, that is, pK(H)(H(3))((Ado)) = -4.02 and pK(H)(H(2))((Ado)) = -1.53; the third one was re-measured as pK(H)(H)((Ado)) = 3.64 +/- 0.02 (25 degrees C; I=0.5 M, NaNO(3)), because it is the main basis for evaluating the intrinsic basicities of N7 and N3. Previously, contradicting results had been published for the micro acidity constant of the (N7)H(+) site; this constant has now been determined in an unequivocal manner, and that of the (N3)H(+) site was obtained for the first time. The micro acidity constants, which describe the release of a proton from an (N)H(+) site under conditions for which the other nitrogen atoms are free and do not carry a proton, decrease in the order pk(N7-N1)(N7(Ado)N1.H)) = 3.63 +/- 0.02 > pk(N7-N1)(H.N7(Ado)N1) = 2.15 +/- 0.15 > pk(N3-N1,N7)(H.N3(Ado)N1,N7) =1.5 +/- 0.3, reflecting the decreasing basicity of the various nitrogen atoms, that is, N1>N7>N3. Application of the above-mentioned microconstants allows one to calculate the percentages (formation degrees) of the tautomers formed for monoprotonated adenosine, H(Ado)(+) , in aqueous solution; the results are 96.1, 3.2, and 0.7% for N7(Ado)N1.H(+), (+)H.N7(Ado)N1, and (+)H.N3(Ado)N1,N7, respectively. These results are in excellent agreement with theoretical DFT calculations. Evidently, H(Ado)(+) exists to the largest part as N7(Ado)N1.H(+) having the proton located at N1; the two other tautomers are minority species, but they still form. These results are not only meaningful for adenosine itself, but are also of relevance for nucleic acids and adenine nucleotides, as they help to understand their metal ion-binding properties; these aspects are briefly discussed. PMID- 21626582 TI - Unidirectional redox-stimulated movement around a C-C single bond. AB - A remarkable challenge for the design of molecular machines is the realization of a synchronized and unidirectional movement caused by an external stimulus. Such a movement can be achieved by a unidirectionally controlled change of the conformation or the configuration. Biphenol derivatives are one possibility to realize a redox-driven unidirectional molecular switch. For this reason, a 4,4' biphenol derivative was fixed to a chiral cyclopeptidic scaffold and stimulated by chemical oxidants and reduction agents. The conformation of the switch was determined by DFT calculations by using B3LYP and the 6-31G* basis set. The switching process was observed by UV and circular dichroism (CD) spectroscopic measurements. Several oxidation agents and various conditions were tested, among which (diacetoxy)iodobenzene (DAIB) in methanol proved to be the best. In this way it was possible to synthesize a redox-stimulated molecular switch with a movement that is part of a rotation around a biaryl binding axis. PMID- 21626583 TI - Investigation of aromaticity and photophysical properties in [18]/[20]pi porphycene derivatives. AB - In this study, we have investigated the relationship between aromaticity and photophysical properties of trifluoromethyl-substituted [18]/[20]pi porphycenes by using theoretical calculations and various spectroscopic methodologies. Interestingly, we have found that the HOMO-LUMO gap of [20]pi porphycene is larger than that of [18]pi porphycene, which is in a sharp contrast with those of typical [4n]/[4n+2]pi porphyrinoids. Based on our observations, we demonstrate that the origin of this contrasting feature of [20]pi porphycene arises from the uniquely large energy splitting between LUMO and LUMO+1 of [18]pi porphycene compared with other aromatic [4n+2]pi porphyrinoids with nearly degenerate LUMO/LUMO+1. Consequently, we can propose that the energy difference between LUMO and LUMO+1 levels of aromatic [4n+2]pi porphyrinoids is an important factor in determining the electronic nature of their corresponding antiaromatic [4n]pi porphyrinoids. Moreover, to the best of our knowledge, this is the first study to illustrate the photophysical properties of porphycenes with [4n]pi electronic circuits. PMID- 21626584 TI - Chiral metallo-supramolecular complexes selectively induce human telomeric G quadruplex formation under salt-deficient conditions. AB - Chiral molecular recognition of human telomeric DNA is important for rational drug design and developing structural probes of G-quadruplexes. Here we report that a chiral supramolecular complex can selectively induce human telomeric G quadruplex formation and discriminate different G-quadruplex sequences under salt deficient conditions studied by circular dichroism (CD), UV meltings, stopped flow spectroscopy, fluorescence resonance energy transfer, enzyme cleavage, and gel electrophoresis. P-enantiomer induced G-quadruplex formation is fast and does not require a large excess of P enantiomer. More importantly, this chiral compound induces loop sequence-dependent G-quadruplex formation. PMID- 21626585 TI - Elansolid A3, a unique p-quinone methide antibiotic from Chitinophaga sancti. PMID- 21626586 TI - Homogeneous nucleophilic radiofluorination and fluorination with phosphazene hydrofluorides. AB - A series of phosphazenium hydrofluorides, P(1)(tBu).[(18/19)F]HF, P(1)(tOct).[(18/19)F]HF, P(2)(Et).[(18/19)F]HF, and P(4)(tBu).[(18/19)F]HF, was synthesized. The radioactive phosphazenium [(18)F]hydrofluorides were obtained by the one-step formation and trapping of gaseous [(18)F]HF with the respective phosphazene bases. The [(19)F] isotopomers were prepared from the corresponding phosphazene bases and Et(3)N.3HF. Under the design of experiment (DoE)-optimized conditions, P(2)(Et).HF and P(4)(tBu).HF fluorinated alkyl chlorides, bromides, and pseudohalides in 76-98% yield, but gave lower yields with iodides and electron-deficient arenes. DoE models showed that fluorination can be performed in glass vessels, and that the reactivity of P(2)(Et).HF and P(4)(tBu).HF is dominated by solvent polarity but is insensitive to water to at least 2 equiv. In contrast, P(1)(tBu).HF and P(1)(tOct).HF were unstable towards autofluorolysis. DFT calculations were performed to rationalize this finding in terms of diminished steric bulk, higher Parr's electrophilicity, and chemical hardness of P(1)(R)H(+). The corresponding radiofluorination reaction gave no valid DoE model but displayed similar substrate scope. High specific activity and excellent radiochemical yields with various pseudohalides (81-91%) suggest that the proposed radiofluorination methodology can complement the current [(18)F]KF/Kryptofix methods, particularly in the areas for which nonpolar reaction conditions are required. PMID- 21626587 TI - Controlled self-assembly of re-engineered insulin by Fe(II). AB - Self-assembly of proteins mediated by metal ions is crucial in biological systems and a better understanding and novel strategies for its control are important. An abiotic metal ion ligand in a protein offers the prospect of control of the oligomeric state, if a selectivity over binding to the native side chains can be achieved. Insulin binds Zn(II) to form a hexamer, which is important for its storage in vivo and in drug formulations. We have re-engineered an insulin variant to control its self-assembly by covalent attachment of 2,2'-bipyridine. The use of Fe(II) provided chemoselective binding over the native site, forming a homotrimer in a reversible manner, which was easily followed by the characteristic color of the Fe(II) complex. This provided the first well-defined insulin trimer and the first insulin variant for which self-assembly can be followed visually. PMID- 21626588 TI - Towards larger polygonal architectures: synthesis and characterization of iron(II)- and ruthenium(II)-bis(terpyridine) metallomacrocycles. PMID- 21626589 TI - One-pot miniemulsion-mediated route to BiOBr hollow microspheres with highly efficient photocatalytic activity. PMID- 21626590 TI - Luminescent peptide labeling based on a histidine-binding iridium(III) complex for cell penetration and intracellular targeting studies. PMID- 21626591 TI - Fast, ligand- and solvent-free synthesis of 1,4-substituted buta-1,3-diynes by Cu catalyzed homocoupling of terminal alkynes in a ball mill. AB - A method for the Glaser coupling reaction of alkynes by using a vibration ball mill has been developed. The procedure avoids the use of ligands and solvents during the reaction. Aryl- and alkyl-substituted terminal alkynes undergo homocoupling if coground with KF-Al(2)O(3) and CuI as a milling auxiliary and catalyst. Furthermore, an alternative protocol has been developed incorporating 1,4-diazabicyclo[2.2.2]octane (DABCO) as an additional base allowing the use of KF-Al(2)O(3) with a lower KF loading. Besides Cu salts, the homocoupling of phenylacetylene is also catalyzed by Ni or Co salts, as well as by PdCl(2). TMS protected phenylacetylene could be directly converted into the homocoupling product after in situ deprotection of the alkyne by fluoride-initiated removal of the trimethylsilyl group. PMID- 21626592 TI - Four iodine-mediated electrophilic cyclizations of rigid parallel triple bonds mapped from 1,8-dialkynylnaphthalenes. AB - Four different types of fused arenes, including fluoranthene, indeno[2,1 a]phenalene, (8H)cyclopenta[a]acenaphthylene, and pyridine[a]acenaphthylene, were efficiently constructed through iodine-mediated electrophilic cyclizations of 1,8 dialkynyl naphthalenes in a single step. Theoretical calculations supported our hypothesis that these reactions had high regioselectivity. Oxidative coupling of the fluoranthene skeleton, followed by aromatization, effectively synthesized perylene derivative 14, which emitted light at 597 nm in dichloromethane with an emission efficiency of 0.81 referred to 5,6,11,12-tetraphenylnaphthacene as a standard. PMID- 21626593 TI - Synthesis of a large functional cage compound based on four Ga-Ga single bonds and its application as an oligoacceptor: on the way to bio-organogallium hybrid molecules. AB - The digallium compound R(2)Ga-GaR(2) (1; R=CH(SiMe(3))(2)) reacts with citracinic acid by the release of two equivalents of bis(trimethylsilyl)methane and the formation of a unique oligofunctional cage compound (2). Four Ga-Ga bonds in a tetrahedral arrangement are bridged by four spacer ligands that are located on the faces of the tetrahedron and bridge the gallium atoms of three different Ga Ga bonds. Four pyridinium groups result from the shift of one of the three acidic protons of four citracinic acid molecules to the nitrogen atoms of the aromatic rings. The N-H groups are arranged in pairs and are capable of acting as chelating acceptors for the coordination of THF molecules (2(THF)(2)) or the nitrogen atoms of 1-deazapurine (3(OEt(2))(4)). In particular, the last reaction verifies the potential applicability of this relatively water- and air-resistant acceptor compound for the generation of bioorganometallic hybrid molecules. PMID- 21626594 TI - Enantioselective double aldol reaction catalyzed by chiral phosphine oxide. PMID- 21626595 TI - Catalytic enantioselective Henry reactions of isatins: application in the concise synthesis of (S)-(-)-spirobrassinin. PMID- 21626596 TI - Ag2O as a new visible-light photocatalyst: self-stability and high photocatalytic activity. PMID- 21626597 TI - Anti-aromatic 16pi porphyrin-metal complexes with meso-alkyl substituents. PMID- 21626598 TI - Direct synthesis of aryl ketones by palladium-catalyzed desulfinative addition of sodium sulfinates to nitriles. PMID- 21626599 TI - Effects of capsaicin on lipid catabolism in 3T3-L1 adipocytes. AB - Capsaicin (8-methyl-N-vanillyl-6-nonenamide) is a pungent ingredient of red peppers, and has been reported to reduce body weight gain and adiposity in rodents. The present study investigated the effects of capsaicin on lipid catabolism in differentiated 3T3-L1 adipocytes. Capsaicin decreased the intracellular lipid content in a concentration-dependent manner. The release of glycerol into the medium was increased by the addition of capsaicin. The mRNA levels of genes involved in lipid catabolism such as hormone sensitive lipase (HSL), carnitine palmitoyl transferase-Ialpha (CPTI-alpha) and uncoupling protein 2 (UCP2) were up-regulated significantly. These results suggest that capsaicin exerts its lipolytic action by increasing the hydrolysis of triacylglycerol in adipocytes, and that these effects are mediated at least partially by regulation of the expression of multiple genes that are involved in the lipid catabolic pathway, such as HSL and CPT-Ialpha, and those involved in thermogenesis such as UCP2. PMID- 21626600 TI - Antifatigue activity of the liposoluble fraction from Acanthopanax senticosus. AB - The crude extract of Acanthopanax senticosus (AS) has been used extensively in Russia, China, Korea and Japan as an adaptogenic agent to fight against stress and fatigue. However, whether the liposoluble fraction possesses antifatigue activity or not is still unclear. A liposoluble fraction was administered orally to mice for 9 days. The swimming time to exhaustion was longer in the treatment groups (22.2 +/- 3.3, 25.5 +/- 4.8 min) than in the control group (13.7 +/- 1.2 min, p < 0.05). The plasma TG (triglyceride) and BUN (blood urea nitrogen) levels in the high dose (500 mg/kg) groups were decreased significantly compared with the control group. Plasma lactate dehydrogenase (LDH) was lower in the treatment groups than in the control group. Chemical analysis from GC/MS revealed that the main components of the liposoluble fraction of AS were saturated fatty acid (12.98%), unsaturated fatty acid (33.13%), unsaturated alcohol (27.46%) and diolefine (15.76%). In conclusion, the liposoluble fraction enhanced the forced swimming capacity of mice by decreasing muscle damage, effectively preventing the increase in BUN concentration and increasing fat utilization. It is proposed that the antioxidant effect may be one of the antifatigue mechanisms of the liposoluble fraction of AS. PMID- 21626601 TI - Intracellular uptake of (-)epicatechin by human erythrocytes as a function of human age. AB - The present studies were carried out to determine the intracellular uptake of ( )epicatechin by erythrocytes during aging in humans. (-)Epicatechin uptake was estimated by performing ethyl acetate extraction while the total antioxidant potential of the (-)epicatechin was estimated in terms of FRAP (ferric reducing ability of plasma) values. A significant (p < 0.001) decrease in (-)epicatechin uptake by human erythrocytes was observed as a function of age. It is hypothesized that the uptake of (-)epicatechin in human erythrocytes has an important role in the regulation of PMRS (plasma membrane redox system) activity during normal human aging. PMID- 21626603 TI - Rethinking herpes simplex virus: the way to oncolytic agents. AB - Oncolytic viruses infect, replicate in and kill cancer cells. HSV has emerged as a most promising candidate because it exerts a generally moderate pathogenicity in humans; it is amenable to attenuation and tropism retargeting; the ample genome provides space for heterologous genes; specific antiviral therapy is available in a worst case scenario. The first strategy to convert HSV into an oncolytic agent consisted in deletion of the gamma(1) 34.5 gene which counteracts the protein kinase R (PKR) response, and of the UL39 gene which encodes the large ribonucleotide reductase subunit. Tumor specificity resided in low PKR activity, and high deoxyribonucleotides content of cancer cells. These highly attenuated viruses have been and presently are in clinical trials with encouraging results. The preferred route of administration has been intratumor or in tissues adjacent to resected tumors. Although the general population has a high seroprevalence of antibodies to HSV, studies in animals and humans demonstrate that prior immunity is not an obstacle to systemic routes of administration, and that oncolytic HSV (o-HSVs) do populate tumors. As the attenuated viruses undergo clinical experimentation, the research pipeline is developing novel, more potent and highly tumor-specific o-HSVs. These include viruses which overcome tumor heterogeneity in PKR level by insertion of anti-PKR genes, viruses which reinforce the host tumor clearance capacity by encoding immune cytokines (IL-12 or granulocyte-macrophage colony-stimulating factor), and non-attenuated viruses fully retargeted to tumor specific receptors. A strategy to generate o-HSVs fully retargeted to human epidermal growth factor receptor-2 (HER-2) or other cancer specific surface receptors is detailed. PMID- 21626604 TI - Simultaneously assessing intended and unintended treatment effects of multiple treatment options: a pragmatic "matrix design". AB - PURPOSE: A key aspect of comparative effectiveness research is the assessment of competing treatment options and multiple outcomes rather than a single treatment option and a single benefit or harm. In this commentary, we describe a methodological framework that supports the simultaneous examination of a "matrix" of treatments and outcomes in non-randomized data. METHODS: We outline the methodological challenges to a matrix-type study (matrix design). We consider propensity score matching with multiple treatment groups, statistical analysis, and choice of association measure when evaluating multiple outcomes. We also discuss multiple testing, use of high-dimensional propensity scores for covariate balancing in light of multiple outcomes, and suitability of available software. CONCLUSION: The matrix design study methods facilitate examination of the comparative benefits and harms of competing treatment choices, and also provides the input required for calculating the numbers needed to treat and for a broader benefit/harm assessment that weighs endpoints of varying severity. PMID- 21626605 TI - Validity of diagnostic codes and liver-related laboratory abnormalities to identify hepatic decompensation events in the Veterans Aging Cohort Study. AB - PURPOSE: The absence of validated methods to identify hepatic decompensation in cohort studies has prevented a full understanding of the natural history of chronic liver diseases and impact of medications on this outcome. We determined the ability of diagnostic codes and liver-related laboratory abnormalities to identify hepatic decompensation events within the Veterans Aging Cohort Study (VACS). METHODS: Medical records of patients with hepatic decompensation codes and/or laboratory abnormalities of liver dysfunction (total bilirubin >= 5.0 g/dL, albumin <= 2.0 g/dL, INR >= 1.7) recorded 1 year before through 6 months after VACS entry were reviewed to identify decompensation events (i.e., ascites, spontaneous bacterial peritonitis, variceal hemorrhage, hepatic encephalopathy, hepatocellular carcinoma) at VACS enrollment. Positive predictive values (PPVs) of diagnostic codes, laboratory abnormalities, and their combinations for confirmed outcomes were determined. RESULTS: Among 137 patients with a hepatic decompensation code and 197 with a laboratory abnormality, the diagnosis was confirmed in 57 (PPV, 42%; 95%CI, 33%-50%) and 56 (PPV, 28%; 95%CI, 22%-35%) patients, respectively. The combination of any code plus laboratory abnormality increased PPV (64%; 95%CI, 47%-79%). One inpatient or >=2 outpatient diagnostic codes for ascites, spontaneous bacterial peritonitis, or variceal hemorrhage had high PPV (91%; 95%CI, 77%-98%) for confirmed hepatic decompensation events. CONCLUSION: An algorithm of 1 inpatient or >= 2 outpatient codes for ascites, peritonitis, or variceal hemorrhage has sufficiently high PPV for hepatic decompensation to enable its use for epidemiologic research in VACS. This algorithm may be applicable to other cohorts. PMID- 21626606 TI - Systematic review of peptic ulcer disease incidence rates: do studies without validation provide reliable estimates? AB - PURPOSE: Incidence rate (IR) estimates for peptic ulcer disease (PUD) vary widely among studies. We conducted a systematic review to quantify and examine the discrepancies. METHODS: Of 4780 articles identified from PubMed and EMBASE databases, 31 published in the last three decades that had reported IRs of PUD in the general population were included. Random effects meta-analysis and meta regression were performed to calculate pooled estimates and to identify sources of heterogeneity. RESULTS: The pooled IR estimate per 1000 person-years was 0.90 (95% confidence interval: 0.78-1.04) for uncomplicated PUD, 0.57 (0.49-0.65) for peptic ulcer bleeding, 0.10 (0.08-0.13) for gastrointestinal perforations, and 3.18 (2.05-4.92) for nonspecific PUD. Within specific outcomes definitions, IR estimates were significantly lower in studies with restriction to hospitalized cases, case validation, and case ascertainment directly from hospital or clinical sources versus computerized health care databases. Younger age, female sex, and later calendar time were also associated with lower PUD incidence. CONCLUSIONS: We found that the IR of uncomplicated PUD was in the order of one case per 1000 person-years in the general population, and that the IR of peptic ulcer complications was around 0.7 cases per 1000 person-years. Comparisons of IR estimates among studies need to take into account disease definition and other study characteristics, particularly whether outcome validation was performed in computerized claims. The use of claims to identify PUD cases might overestimate the IR by around 45%. PMID- 21626607 TI - Nationwide register-based surveillance system on drugs and pregnancy in Finland 1996-2006. AB - PURPOSE: The aim of this population-based nationwide drugs and pregnancy surveillance system was to get basic information on the use of prescribed drugs during pregnancy and to achieve more detailed information on drug exposure outcome associations with data obtained from the Finnish national health registers. METHODS: The data are based on information on all pregnancies ending in births (n = 632,629) or termination of pregnancy (TOP) (n = 117,255) in Finland between 1996 and 2006. The data containing information on maternal background factors, chronic diseases and drug purchases during pregnancy, born children, induced abortions, perinatal health and major congenital malformations were obtained from the Finnish national health registers. Information from the different registers was merged through record linkages based on unique personal identification numbers. Statistical analyses were conducted between the exposed and unexposed for individual drugs or Anatomical Therapeutic Chemical (ATC) classification drug groups in multivariate logistic regression including potential confounding factors. RESULTS: Nearly half of parturients and almost every third woman with TOP purchased drugs at least once 1 month prior to pregnancy and/or during pregnancy. The most frequent chronic diseases during pregnancy were asthma, hypothyroidism, epilepsy, rheumatoid arthritis and diabetes. Previously known causal connections between maternal chronic diseases and/or medication and perinatal health risks were already seen in the ATC-group level analysis. CONCLUSIONS: Comprehensive and detailed information for pharmacoepidemiological research on the effects of drug use during pregnancy in a cohort setting is possible with long-term and cumulative data collection. PMID- 21626608 TI - Valuing states from multiple measures on the same visual analogue sale: a feasibility study. AB - Previous methods of empirical mapping involve using regressions on patient or general population self-reported data from datasets involving two or more measures. This approach relies on overlap in the descriptive systems of the measures and assumes it is appropriate to use different measures on the same population, which may not always be the case. This paper presents a feasibility study for a new approach to mapping between preference-based measures (PBM) using general population visual analogue scale (VAS) values as a common yardstick. We use data from a valuation study of 502 members of the UK general population, where, using ranking and VAS tasks, interviewees simultaneously valued health states defined by three of six PBM: EQ-5D (generic), SF-6D (generic), HUI2 (generic for children and adults), AQL-5D (asthma specific), OPUS (social care specific) and ICECAP (capabilities). Regression techniques are used to estimate the relationship between these VAS values and the original value set (i.e. 'tariff'). These results are subsequently used to estimate the relationship between all six PBM to enable 'value-based mapping' between measures. This new method of mapping potentially has a useful role in evidence synthesis and cross programme comparisons in studies using different measures. PMID- 21626609 TI - Screening for distress, the 6th vital sign: where are we, and where are we going? PMID- 21626611 TI - The HADS and the DT for screening psychosocial distress of cancer patients in Taiwan. AB - PURPOSE: This study assesses the sensitivity and specificity of Mandarin versions of two psychosocial screening tools for adjustment, anxiety and depressive disorders: the Hospital Anxiety and Depression Scale (HADS), and the Distress Thermometer (DT). METHODS: The two scales were used to screen 103 consecutive cancer patients seen for psychiatric evaluation at KF-SYSCC between May and November 2004 prior to their psychiatric interviews. Each scale was tested against clinical psychiatric diagnoses based on the Diagnostic and Statistical Manual of Mental Disorders, 4th edition for their sensitivity and specificity. RESULTS: For the Mandarin version of the DT, receiver operating characteristic (ROC) analyses identified a DT score of 4 as the optimal cut-off, with sensitivity and specificity of 98 and 73%, respectively. For the Mandarin version of the HADS, ROC identified a score of 9 and 8 as the optimal cutoffs for the respective anxiety and depression subscales (HADS-a and HADS-d), with sensitivities and specificities of 84 and 73, 72 and 86%, respectively. For the full scale of the HADS (HADS-t), 15 was identified as the optimal cutoff, which yielded sensitivity and specificity of 84 and 68%, respectively. Using the frequency table, the concordance rate of the two scales was found to be 72-80% based on the above optimal cut-offs. CONCLUSION: The Mandarin versions of the HADS and the DT are efficacious for screening anxiety and depression for our population. Compared with the HADS-t, the DT appears to have not only higher sensitivity, but also higher specificity. PMID- 21626612 TI - Screening for psychological and physical distress in a cancer inpatient treatment setting: a pilot study. AB - OBJECTIVE: Although several large distress studies have been conducted in outpatient oncology treatment settings, there are few, if any, studies that describe the incidence of psychological and physical distress in the inpatient oncology treatment setting. This pilot study hypothesized that inpatients experience levels of psychological distress that are equivalent to or greater than those reported in large outpatient studies. The study also sought to contribute to physical and psychological symptom distress incidence data. METHODS: Using two measures designed to screen for psychological and physical distress, the Brief Symptom Inventory-18 and the M.D. Anderson Symptom Inventory, 52 oncology patients were screened within 24 h of admission to a hospital oncology unit. RESULTS: Nearly two-thirds of the inpatient respondents reported levels of psychological distress that were not only within the clinically significant range but also at a rate that was nearly double that found in the outpatient distress screening studies that have been conducted to date. The physical symptoms most often rated at the severe level included fatigue (34.6%), pain (27.4%), and dry mouth (25.5%). CONCLUSIONS: Failure to screen for psychological and physical symptom distress has lead to under recognition of a population that may have higher levels of distress than their outpatient counterparts and could potentially lead to uneven access to biopsychosocial support. Results support the notion that routinely screening for psychological and physical distress should become a first step in the assessment of the biopsychosocial needs of people receiving inpatient treatment for cancer. PMID- 21626610 TI - Distress screening in allogeneic hematopoietic stem cell (HSCT) caregivers and patients. AB - Family caregivers of allogeneic hematopoietic stem cell transplant (HSCT) patients are at risk for experiencing significant psychological distress yet screening caregivers has not been well studied. OBJECTIVE: This analysis explored the psychometric characteristics of the Distress Thermometer (DT) by examining its relationship, sensitivity, and specificity relative to the Brief Symptom Inventory 18 (BSI-18) and the Multidimensional Fatigue Symptom Inventory (MFSI) in a sample of allogeneic HSCT caregivers and patients. METHODS: Longitudinal data were drawn from an ongoing intervention study for HSCT caregivers and patients. Data from one hundred and fifty-six English-speaking adults where patients (n = 65) were receiving their first allogeneic HSCT with at least one adult caregiver (n = 91) were eligible for this analysis. Study questionnaires were administered at baseline, initial discharge, and 6 weeks following discharge. RESULTS: Construct validity was supported by significant relationships (p<0.001) between the DT and the BSI-18 GSI and the MFSI-Emotional subscales for caregivers and patients. The diagnostic utility of the DT for patients was good (AUC = 0.85+/-0.05, p = 0.001), while for caregivers it was poor (AUC = 0.61+/ 0.08, p = 0.28). A DT cut point of 5 was supported for patients (sensitivity = 1.0, specificity = 0.68), while for caregivers there was less confidence (sensitivity = 0.70, specificity = 0.52). Caregivers and patients reporting a higher number of problems had a greater level of distress (p<0.001). CONCLUSIONS: These findings support the validity of the DT in screening for distress in HSCT caregivers and patients. Although the diagnostic utility of the DT for HSCT caregivers may be limited, understanding factors associated with distress can guide practice for this understudied population. PMID- 21626614 TI - Upconverting nanoparticles. AB - Upconversion (UC) refers to nonlinear optical processes in which the sequential absorption of two or more photons leads to the emission of light at shorter wavelength than the excitation wavelength (anti-Stokes type emission). In contrast to other emission processes based on multiphoton absorption, upconversion can be efficiently excited even at low excitation densities. The most efficient UC mechanisms are present in solid-state materials doped with rare earth ions. The development of nanocrystal research has evoked increasing interest in the development of synthesis routes which allow the synthesis of highly efficient, small UC particles with narrow size distribution able to form transparent solutions in a wide range of solvents. Meanwhile, high-quality UC nanocrystals can be routinely synthesized and their solubility, particle size, crystallographic phase, optical properties and shape can be controlled. In recent years, these particles have been discussed as promising alternatives to organic fluorophosphors and quantum dots in the field of medical imaging. PMID- 21626613 TI - Psychodynamic group psychotherapy: impact of group length and therapist professional characteristics on development of therapeutic alliance. AB - OBJECTIVES: Little research has been done on therapeutic alliance in group psychotherapy, especially the impact of treatment duration and therapist professional characteristics. METHODS: Therapeutic alliance was rated by patients on the Working Alliance Inventory-Short Form at three time points (sessions 3, 10 and 17) in a randomized controlled trial of short-term and long-term psychodynamic group psychotherapy. As predictors we selected therapist clinical experience and length of didactic training, which have demonstrated ambiguous results in previous research. Linear latent variable growth curve models (structural equation modeling) were developed for the three Working Alliance Inventory-Short Form subscales bond, task and goal. RESULTS: We found a significant variance in individual growth curves (intercepts and slopes) but no differential development due to group length. Longer therapist formal training had a negative impact on early values of subscale task in both treatments. There was an interaction between length of the therapists' clinical experience and group length on early bond, task and goal: therapists with longer clinical experience were rated lower on initial bond in the long-term group but less so in the short-term group. Longer clinical experience influenced initial task and goal positively in the short-term group but was unimportant for task or significantly negative for goal in the long-term group. CONCLUSION: There was no mean development of alliance, and group length did not differentially impact the alliance during 6 months. Early ratings of the three Working Alliance Inventory Short Form subscales partly reflected different preparations of patients in the two group formats, partly therapist characteristics, but more research is needed to see how these aspects impact alliance development and outcome. Therapists should pay attention to all three aspects of the alliance, when they prepare patients for group therapy. KEY PRACTITIONER MESSAGE: In psychodynamic groups, length of therapy does not differentiate the overall level or the development of member-leader alliance. Within psychodynamic groups, each individual appear to have their unique perception of the member-leader alliance. Therapists with longer formal psychotherapy training may be less successful in establishing early agreement with patients on the tasks of psychodynamic group psychotherapy. Patients perceive a somewhat lower degree of early emotional bonding with the more clinically experienced therapists in long-term psychodynamics groups. Therapists with more clinical experience may contribute to a stronger degree of initial agreement with patients on the tasks and goals of short-term group psychotherapy. PMID- 21626615 TI - Electrospinning of highly electroactive carbon-coated single-crystalline LiFePO4 nanowires. PMID- 21626616 TI - Determination of molecular surface structure, composition, and dynamics under reaction conditions at high pressures and at the solid-liquid interface. AB - In the last two decades, surface-science experiments and techniques have been developed to focus on obtaining molecular information under reaction conditions at high pressures (near or above 1 bar) and liquid interfaces. This Minireview describes the results of these studies obtained by surface-sensitive laser spectroscopies, scanning tunneling microscopy, and X-ray spectroscopies usually practiced at a synchrotron light source. The use of model surfaces, single crystals, and monodisperse nanoparticles with variable size (1-10 nm) and shape facilitates meaningful interpretation of the experimental data. These methods allow evaluation of the molecular structures of intermediates, oxidation states of metals, and mobility of adsorbants. New techniques that are likely to make major contributions to the investigation of surfaces under reaction conditions are also discussed. PMID- 21626617 TI - An efficient and low-cost method for the purification of colloidal nanoparticles. PMID- 21626618 TI - Memory effects in molecular films of free-standing rod-shaped ruthenium complexes on an electrode. PMID- 21626619 TI - Electrostatic control of bioactivity. PMID- 21626620 TI - C1-symmetric Rh/Phebox-catalyzed asymmetric alkynylation of alpha-ketoesters. PMID- 21626621 TI - Chelating carboxylic acid amides as robust relay protecting groups of carboxylic acids and their cleavage under mild conditions. PMID- 21626622 TI - Rhodium(I)-catalyzed ene-allene-allene [2+2+2] cycloadditions: stereoselective synthesis of complex trans-fused carbocycles. PMID- 21626623 TI - Cathodic corrosion: a quick, clean, and versatile method for the synthesis of metallic nanoparticles. PMID- 21626624 TI - Visual chiral recognition through enantioselective metallogel collapsing: synthesis, characterization, and application of platinum-steroid low-molecular mass gelators. PMID- 21626625 TI - A butterfly-shaped amphiphilic molecule: solution-transferable and free-standing bilayer films for organic transistors. PMID- 21626626 TI - Radical derivatives of insoluble La@C74: X-ray structures, metal positions, and isomerization. PMID- 21626627 TI - Supercapacitor electrodes with high-energy and power densities prepared from monolithic NiO/Ni nanocomposites. PMID- 21626628 TI - Cyclic polymers with pendent carbazole units: enhanced fluorescence and redox behavior. PMID- 21626629 TI - A high-efficiency panchromatic squaraine sensitizer for dye-sensitized solar cells. PMID- 21626630 TI - Palladium-mediated annulation of vinyl aziridines with Michael acceptors: stereocontrolled synthesis of substituted pyrrolidines and its application in a formal synthesis of (-)-alpha-kainic acid. PMID- 21626631 TI - Protein-DNA recognition triggered by a DNA conformational switch. PMID- 21626632 TI - Bioinspired functionalization of silica-encapsulated yeast cells. PMID- 21626633 TI - Controlled self-assembly of rodlike bacterial pili particles into ordered lattices. PMID- 21626634 TI - A multicomponent coupling reaction induced by insertion of arynes into the C=O bond of formamide. PMID- 21626635 TI - Directed evolution of an error-prone T7 DNA polymerase that attenuates viral replication. AB - Experimental evidence exists that RNA viruses replicate with extremely high mutation rates that result in significant genetic diversity. The diverse nature of viral populations allows rapid adaptation to dynamic environments, and evolution of resistances to vaccines as well as antiviral substances. For DNA viruses that replicate at much greater fidelities, as yet, neither diverse structures in the population nor their responses to increased mutation rates have been sufficiently described. By using the example of DNA bacteriophage T7, we describe the identification of virus-specific DNA polymerase variants with decreased replication fidelities, and their impact on the efficiency of the viral infection cycle. PMID- 21626636 TI - Sonochemical synthesis of DNA nanospheres. PMID- 21626637 TI - A fast and precise approach for computational saturation mutagenesis and its experimental validation by using an artificial (betaalpha)8-barrel protein. AB - We present a computational saturation mutagenesis protocol (CoSM) that predicts the impact on stability of all possible amino acid substitutions for a given site at an internal protein interface. CoSM is an efficient algorithm that uses a combination of rotamer libraries, side-chain flips, energy minimization, and molecular dynamics equilibration. Because CoSM considers full side-chain and backbone flexibility in the local environment of the mutated position, amino acids larger than the wild-type residue are also modeled in a proper manner. To assess the performance of CoSM, the effect of point mutations on the stability of an artificial (betaalpha)(8)-barrel protein that has been designed from identical (betaalpha)(4)-half barrels, was studied. In this protein, position 234(N) is a previously identified stability hot-spot that is located at the interface of the two half barrels. By using CoSM, changes in protein stability were predicted for all possible single point mutations replacing wild-type Val234(N). In parallel, the stabilities of 14 representative mutants covering all amino acid classes were experimentally determined. A linear correlation of computationally and experimentally determined energy values yielded an R(2) value of 0.90, which is statistically significant. This degree of coherence is stronger than the ones we obtained for established computational methods of mutational analysis. PMID- 21626638 TI - Insights into the mechanistic role of the [Fe4S4] cubane in the A-cluster {[Fe4S4]-(SR)-[NipNid]} of acetyl-coenzyme A synthase. PMID- 21626639 TI - Identification and characterization of the althiomycin biosynthetic gene cluster in Myxococcus xanthus DK897. PMID- 21626640 TI - Note on "The electrochemical promotion of ethylene oxidation at a Pt/YSZ catalyst". PMID- 21626641 TI - Activation/inhibition effects during the coelectrodeposition of PtAg nanoparticles: application for ORR in alkaline media. AB - PtAg bimetallic nanoparticles for oxygen reduction reaction (ORR) in alkaline media were prepared by pulse electrodeposition (PED). During PED the reduction of Ag(+) ions predominates, thus an increased Ag content in the co-deposit is accomplished. The mechanism for this anomalous co-deposition was elucidated by potential pulse experiments, which revealed that nuclei formation mainly occurs via the reduction of Pt(2+) ions. The growth of the particles is diffusion controlled leading to the formation of a Ag shell covering a PtAg alloyed region. However, the shell is not growing homogeneously on the PtAg alloy. Hence, regions of the PtAg alloy are exposed, which exhibit an enhanced ORR activity compared to a pure Ag surface. PMID- 21626642 TI - Surfactin self-assembles into direct and reverse aggregates in equilibrium and performs selective metal cation extraction. AB - On tie-lines between water-rich and alkane-rich solutions, it is shown via scattering experiments that natural lipopeptide surfactin self-assembles into direct and reverse micelles in equilibrium. Elongated direct micelles in the aqueous phase are present together with small reverse globular aggregates in the organic phase. These latter are made from hydrated surfactant without any "water pool" in the organic phase. The resulting biphasic system is used for liquid liquid extraction of model metal cations. It is efficient with iron but not with copper or neodymium. Competitive extractions show high selectivity towards iron. PMID- 21626643 TI - Long-term stable silver subsurface ion-exchanged glasses for SERS applications. AB - We report on the formation of silver subsurface ion-exchanged metal oxide (silver SIMO) glasses and their surface-enhanced Raman scattering (SERS) activity. The samples were prepared by a combined thermal and chemical three-step methodology and characterized by transmission electron microscopy (TEM), atomic force microscopy (AFM), environmental electron scanning microscopy (ESEM), and UV/Vis spectroscopy. This unique method provides SERS substrates with protection against contamination and strong, reliable and reproducible SERS enhancement. The Raman enhancement factors of the long-term stable SIMO glasses were estimated to approximately 10(7). PMID- 21626644 TI - Natural orbital functional theory and reactivity studies of diradical rearrangements: ethylene torsion as a case study. AB - Natural orbital functional theory (NOFT) in its PNOF3 and PNOF4 implementations is used to investigate the potential energy surface (PES) of ethylene torsion. This rearrangement is taken as a case study to illustrate the satisfactory performance of the Piris Natural Orbital Functional (PNOF) toward rearrangements sensible to non-dynamical electron correlation or near-degeneracy effects. We show that the fulfillment of the G-positivity condition, as implemented in PNOF4, is key to yield a correct description of structures with significant diradical character. We have found that PNOF4 yields an accurate description of these delicate PES and predicts the correct trends of the occupation numbers as compared with multiconfigurational wavefunction methods. PMID- 21626645 TI - Dealloying Ag-Al alloy to prepare nanoporous silver as a substrate for surface enhanced Raman scattering: effects of structural evolution and surface modification. AB - Sensitive detection of molecules by using the surface-enhanced Raman scattering (SERS) technique depends on the nanostructured metallic substrate and many efforts have been devoted to the preparation of SERS substrates with high sensitivity, stability, and reproducibility. Herein, we report on the fabrication of stable monolithic nanoporous silver (NPS) by chemical dealloying of Ag-Al precursor alloys with an emphasis on the effect of structural evolution on SERS signals. It was found that the dealloying conditions had great influence on the morphology (the ligament/pore size) and the crystallization status, which determined the SERS signal of rhodamine 6G on the NPS. NPS with small pores, low residual Al, and perfect crystallization gave high SERS signals. A high enhancement factor of 7.5 * 10(5) was observed on bare NPS obtained by dealloying Ag(30)Al(70) in 2.5 wt % HCl at room temperature followed by 15 min aging at around 85 degrees C. After coating Ag nanoparticles on the NPS surface, the enhancement factor increased to 1.6 * 10(8) owing to strong near-field coupling between the ligaments and nanoparticles. PMID- 21626647 TI - Phenethyl isothiocyanate promotes immune responses in normal BALB/c mice, inhibits murine leukemia WEHI-3 cells, and stimulates immunomodulations in vivo. AB - Enhanced cruciferous vegetable consumption is associated with the reduction of cancer incidence as shown in epidemiological studies. Phenethyl isothiocyanate (PEITC), one of the important compounds in cruciferous vegetables, has been shown to induce apoptosis in many types of human cancer cell lines, but there is no available information addressing the effects on normal and leukemia mice in vivo. The purpose of this study is to focus on the in vivo effects of PEITC on immune responses of normal and WEHI-3 leukemia BALB/c mice in vivo. Influences of PEITC on BALB/c mice after intraperitoneal (i.p.) injection with WEHI-3 cells and normal mice were investigated. In normal BALB/c mice, PEITC did not affect the body weight when compared to the olive oil treated animals. Moreover, PEITC promoted phagocytosis by macrophages from peripheral blood mononuclear cells (PBMC) and peritoneal cavity, increased the levels of CD11b and Mac-3, decreased the level of CD19 and promoted natural killer (NK) cell cytotoxic activity, but it did not alter the level of CD3. Also, PEITC enhanced T cell proliferation after concanavalin A (Con A) stimulation. Otherwise, PEITC increased the body weight, but decreased the weight of liver and spleen as compared to the olive oil treated WEHI-3 leukemia mice. PEITC also increased the level of CD19, decreased the levels of CD3 and Mac-3 rather than influence in the level of CD11b, suggesting that the differentiation of the precursor of macrophages and T cells was inhibited, but the differentiation of the precursor of B cells was promoted in leukemia mice. Furthermore, PEITC enhanced phagocytosis by monocytes and macrophages from PBMC and peritoneal cavity, and also promoted the NK cell cytotoxic activity in comparison with the group of leukemia mice. Based on these observations, the biological properties of PEITC can promote immune responses in normal and WEHI-3 leukemia mice in vivo. (c) 2011 Wiley Periodicals, Inc. Environ Toxicol, 2013. PMID- 21626648 TI - Retinoic acid influences the embryoid body formation in mouse embryonic stem cells by induction of caspase and p38 MAPK/JNK-mediated apoptosis. AB - Although all-trans retinoic acid (RA), the oxidative metabolite of vitamin A, is essential for normal development, high levels are teratogenic in many species. RA results in immediate effects on the preimplantation embryo and on blastocyst development in vitro and in vivo. To further elucidate the cellular mechanisms of early postimplantation embryo development induced by RA, we present an embryonic cell line, B5, as a candidate system for the investigation of these processes. We used undifferentiated ES cells as the model, which is from the undifferentiated status to differentiated status [embryoid body (EB) formation] mimicking postimplantation embryo development (egg-cylinder stage of embryo formation) to clarify the cellular mechanism of action of RA in the implanted blastocysts and cell apoptosis following the series of exposures to differing RA concentrations. Using an in vitro model, we identified the impact of RA on undifferentiated embryonic stem (ES) cells, including inhibition of cell proliferation and induction of cell apoptosis. JNK, P-38 and caspase activation were shown in the nature of RA-triggered apoptotic signaling in ES cells. The carry-on influences of RA on the ES cell were shown in the formation of EB from the pretreated ES cells. RA resulted in apparent impact on undifferentiated ES cells in vitro, with increased numbers of apoptotic cells initially and inhibited cell proliferation, which led to decreased size of EB. The process of EB formation (mimicking the early postimplantation embryo development) is regulated by RA-induced apoptosis through the activation of caspase and P38 MAPK/JNK pathway. PMID- 21626649 TI - Excessive fluoride induces endoplasmic reticulum stress and interferes enamel proteinases secretion. AB - Protein retention in the enamel layer during tooth formation is well known to be associated with dental fluorosis but the underlying mechanism is unclear. The functions of the endoplasmic reticulum (ER) correlate directly with secreted protein metabolism. We used an ameloblast-derived cell line to determine whether excessive amounts of fluoride cause ER stress, and whether this interferes with the secretion of enamel matrix proteinases. ER stress activates a signaling network called the unfolded protein response (UPR). Here, we used real-time RT PCR and immunofluorescence to study the effect of fluoride on the expression, translation, and secretion of UPR transcription factors in ameloblast-like cells. Measurement of both the gene and protein expression of UPR transcription factors indicated that high-dose fluoride increases the expression of UPR transcription factors in a dose-dependent manner. We also used ELISA to detect and quantify the enamel proteinases secreted by ameloblasts. We found a corresponding decrease in extracellular secretion of the enamel proteinases matrix metalloproteinase-20 and kallikrein-4, after exposure to fluoride. Furthermore, correlation analysis indicated that the expression of UPR transcription factors showed a strong inverse correlation with that of enamel proteinases. The results suggest that high-dose fluoride initiates an ER stress response in ameloblasts and induces the UPR, which interferes with the synthesis and secretion of enamel proteinases. Taken together, these results suggest that excessive ingestion of fluoride during tooth formation can decrease the secretion of proteinases, thus causing protein retention in the enamel layer, indicating that the ER stress response may be responsible for dental fluorosis. PMID- 21626650 TI - Influence of increasing temperature and salinity on herbicide toxicity in estuarine phytoplankton. AB - Ecological risk assessments are, in part, based on results of toxicity tests conducted under standard exposure conditions. Global climate change will have a wide range of effects on estuarine habitats, including potentially increasing water temperature and salinity, which may alter the risk assessment of estuarine pollutants. We examined the effects of increasing temperature and salinity on the toxicity of common herbicides (irgarol, diuron, atrazine, and ametryn) to the phytoplankton species Dunaliella tertiolecta. Static 96-h algal bioassays were conducted for each herbicide under four exposure scenarios: standard temperature and salinity (25 degrees C, 20 ppt), standard temperature and elevated salinity (25 degrees C, 40 ppt), elevated temperature and standard salinity (35 degrees C, 20 ppt), and elevated temperature and elevated salinity (35 degrees C, 40 ppt). The endpoints assessed were algal cell density at 96 h, growth rate, chlorophyll a content, lipid content, and starch content. Increasing exposure temperature reduced growth rate and 96-h cell density but increased the cellular chlorophyll and lipid concentrations of the control algae. Exposure condition did not alter starch content of control algae. Herbicides were found to decrease growth rate, 96 h cell density, and cellular chlorophyll and lipid concentrations, while starch concentrations increased with herbicide exposure. Herbicide effects under standard test conditions were then compared with those observed under elevated temperature and salinity. Herbicide effects on growth rate, cell density, and starch content were more pronounced under elevated salinity and temperature conditions. To encompass the natural variability in estuarine temperature and salinity, and to account for future changes in climate, toxicity tests should be conducted under a wider range of environmental conditions. PMID- 21626651 TI - Developmental exposure to decabrominated diphenyl ether (BDE-209): effects on sperm oxidative stress and chromatin DNA damage in mouse offspring. AB - Polybrominated diphenyl ethers (PBDEs) are used as brominated flame retardants and have been found in human milk in recent years. This study investigates whether prenatal exposure to decabrominated diphenyl ether (BDE-209) induces sperm dysfunction in male offspring. Pregnant CD-1 mice were gavaged once daily with corn oil (control), 10, 500, and 1500 mg kg(-1) body weight of BDE-209 from day 0 of gestation to day 17. The outcomes of male reproductive parameters were assessed on postnatal day 71. Anogenital distance, sperm-head abnormalities, and testicular histopathology were significantly affected in male offspring prenatally exposed to 1500 mg kg(-1). Significant increases in the tendency for sperm DNA denaturation (alphaT) induction and the DNA fragmentation index (DFI) were found in those exposed to 10, 500, and 1500 mg kg(-1) (P < 0.05). We observed a significant increase of sperm hydrogen peroxide (H(2)O(2)) generation in the 10 and 1500 mg/kg/day groups compared to the control group (P < 0.05). Although our findings suggested that the mechanisms underlying BDE-209-induced sperm DNA damage and H(2)O(2) generation might not be represented as a dose response relationship, we found that the greater the excess production of sperm H(2)O(2), the greater the sperm alphaT (r = 0.65, P = 0.0155) and DFI (r = 0.53, P = 0.002). In conclusion, developmental exposure to BDE-209 induced sperm-head abnormality, oxidative stress, chromatin DNA damage, and testicular histopathological changes. These findings suggest that BDE-209-induced male reproductive effects might involve the formation of sperm H(2)O(2) which attacks nucleic acids via H(2)O(2) generation. PMID- 21626652 TI - Inhibition of PP2A and the consequent activation of JNK/c-Jun are involved in tributyltin-induced apoptosis in human amnionic cells. AB - Tributyltin (TBT), a highly toxic environmental contaminant, has been shown to induce mitochondrial-dependent apoptosis in several mammalian cells. However, the upstream signal transduction pathways involved in TBT-induced apoptosis are still not fully elucidated. In this study, the protein phosphatase (PP) 2A, microtubule organization, and mitogen-activated protein kinases (MAPKs), including JNK, p38 and their downstream transcription factors, c-Jun and ATF-2, respectively, were investigated in human amnionic cells treated by TBT. Furthermore, the activation of procaspase-3 after blocking either one of these MAPK pathways was also observed. The results showed that TBT effectively induced apoptosis characterized by caspase-3 activation. In apoptotic cells, the inhibition of PP2A activity and microtubule depolymerization was detected. Additionally, JNK and p38, as well as their downstream targets, c-Jun and ATF-2, were activated. Moreover, a JNK inhibitor, but not p38 inhibitor, significantly reduced caspase-3 activation. It can be concluded that the inhibition of PP2A may (1) play as a role in the activation of JNK and c-Jun and the concomitant promotion of microtubule depolymerization and (2) lead to the activation of caspase-3 in TBT-induced apoptotic cells. The results of this study suggest a critical role of PP2A in the TBT toxicity mechanism. PMID- 21626653 TI - The impact of unsuccessful percutaneous coronary intervention on short- and long term prognosis in STEMI and NSTEMI. AB - OBJECTIVES: To compare the impact of the efficacy of percutaneous coronary intervention (PCI) on prognosis in ST and non-ST elevation myocardial infarction (STEMI and NSTEMI) patients with respect to infarct-related artery (IRA). BACKGROUND: The significance of the efficacy of PCI in STEMI and NSTEMI depending on the type of IRA has yet to be clarified. METHODS: Study population consisted of 2,179 STEMI and 554 NSTEMI consecutive patients treated with urgent PCI. The efficacy of PCI (TIMI [thrombolysis in myocardial infarction] 3 vs. TIMI < 3) was assessed with regard to the type of IRA (left anterior descending artery, circumflex artery [Cx] or right coronary artery). The mean follow-up was 37.5 months. RESULTS: The rate of unsuccessful PCI was similar in STEMI and NSTEMI irrespectively of IRA (14.1 vs. 17.7%; P = 0.062). In STEMI, unsuccessful PCI was associated with significantly higher early (23.1 vs. 5.6%; P < 0.001) and late (29.9 vs. 12.8%; P < 0.001) mortality regardless of IRA. In NSTEMI, the inefficacious PCI significantly increased early (19.0% vs. 0.9%; P < 0.001) and late (27.3% vs. 6.3%; P < 0.001) mortality only in patients with Cx-related infarction. Unsuccessful PCI of IRA was an independent risk factor for death in STEMI (HR 1.64; P < 0.05), but not in NSTEMI (P = 0.64). Further analysis showed that whilst unsuccessful PCI of any vessel in STEMI is an independent risk factor for death, in NSTEMI this applies to unsuccessful PCI of Cx only. CONCLUSIONS: The significance of unsuccessful PCI of IRA seems to be different in STEMI and NSTEMI. Unsuccessful PCI is an independent risk factor for death in STEMI regardless of IRA and in NSTEMI with the involvement of Cx. PMID- 21626654 TI - Evaluation of toxicity and genotoxic effects of spinosad and deltamethrin in Drosophila melanogaster and Bactrocera oleae. AB - BACKGROUND: The insecticides spinosad and deltamethrin are being increasingly used in pest management programmes. In order to assess further their toxic effects to target and non-target insect species, an evaluation was made of their insecticidal profile on Bactrocera oleae (Rossi) and Drosophila melanogaster (Meig.). Moreover, possible genotoxic effects of the two pesticides were investigated using the somatic mutation and recombination test (SMART) in D. melanogaster. RESULTS: Both insecticides were highly effective against B. oleae, exhibiting similar LC(50) values. Moreover, they were found to be more effective against Bactrocera than against Drosophila adults. However, spinosad was significantly more toxic than deltamethrin to D. melanogaster. The results showed a lack of genotoxic activity of both insecticides under the in vivo experimental procedure employed, at least at applied doses. CONCLUSION: The present study provides information for lethal and sublethal effects of spinosad and deltamethrin against a target and a non-target species. Both insecticides can exert high toxicity to B. oleae when adults are exposed even to very low doses for long periods of time. The results contribute to the database on the genotoxic potential of spinosad and deltamethrin, suggesting a safety profile for both insecticides. PMID- 21626655 TI - Organic superconductors. AB - The present status of organic superconductors of charge-transfer (CT) type based on donor molecules is reviewed. Along with the superconducting phases of such materials and also of oxide superconductors, reside spin-ordered phases such as spin-density wave (SDW) and antiferromagnetic (AF) phases. We briefly describe the recent development of superconductors having a superconducting phase next to a spin-disorder state (quantum spin liquid state). In addition to the CT type superconductors, there are a few single-component superconductors under high pressure. PMID- 21626656 TI - Quantification of protein immobilization on substrates for cellular microarray applications. AB - Cellular microarray developments and its applications are the next step after DNA and protein microarrays. The choice of the surface chemistry of the substrates used for the implementation of this technique, that must favor proper protein immobilization while avoiding cell adhesion on the nonspotted areas, presents a complex challenge. This is a key issue since usually the best nonfouling surfaces are also the ones that retain immobilized the smallest amounts of printed protein. To quantitatively assess the amount of protein immobilization, in this study several combinations of fluorescently labeled fibronectin (Fn*) and streptavidin (SA*) were microspotted, with and without glycerol addition in the printing buffer, on several substrates suitable for cellular microarrays. The substrates assayed included chemically activated surfaces as well as Poly ethylene oxide (PEO) films that are nonfouling in solution but accept adhesion of proteins in dry conditions. The results showed that the spotted Fn* was retained by all the surfaces, although the PEO surface did show smaller amounts of immobilization. The SA*, on the other hand, was only retained by the chemically activated surfaces. The inclusion of glycerol in the printing buffer significantly reduced the immobilization of both proteins. The results presented in this article provide quantitative evidence of the convenience of using a chemically activated surface to immobilize proteins relevant for cellular microarray applications, particularly when ECM proteins are cospotted with smaller factors which are more difficult to be retained by the surfaces. PMID- 21626657 TI - Hydroxyapatite scaffolds infiltrated with thermally crosslinked polycaprolactone fumarate and polycaprolactone itaconate. AB - In this work, two unsaturated derivatives of polycaprolactone (PCL), polycaprolactone fumarate (PCLF), and polycaprolactone itaconate (PCLI), have been synthesized and used as an infiltrating polymer to improve the mechanical properties of brittle hydroxyapatite (HA) scaffolds. PCLF and PCLI were first synthesized through polyesterification of the low molecular weight PCL diols with fumaryl chloride and itaconyl chloride respectively, and then characterized by Fourier transform infrared spectroscopy, nuclear magnetic resonance spectroscopy, gel permeation chromatography, and differential scanning calorimetry analysis. HA scaffolds were sintered using a foam replication technique, with porosity of about 60%. Polymer-HA composites were obtained by infiltrating the HA scaffolds with PCLF and PCLI solution (12.5 and 30 w/v in dichloromethane) followed by thermal crosslinking. The polymer infiltrated HA scaffolds were characterized by scanning electron microscopy, porosimetry, and gravimetrical analysis. The polyesterification reaction of PCL diols with fumarate chloride was more efficient than itaconyl chloride and dependent upon the molecular weight of the initial PCL precursor; the resultant PCLF demonstrated a degree of substitution of 1.2, 4.2, and 2.7 times higher than PCLIs. Polymer infiltration improved the compressive strength of the HA scaffolds, and based upon the type of macromer (PCLF or PCLI) and also their concentration in infiltrating solution (12.5 or 30 w/v %) compressive strength increased about 14-328%. In all studied samples, the reinforcement effect of PCLF infiltration was higher than PCLI. The macromers and their corresponding infiltrated HA scaffolds did not show any significant cytotoxicity toward human primary osteogenic sarcoma cell (G92 cell lines), in vitro. PMID- 21626658 TI - Compositional control of poly(ethylene glycol) hydrogel modulus independent of mesh size. AB - Poly(ethylene glycol) (PEG) hydrogels are of great interest in tissue engineering because of their established biocompatibility, high permeability, and tunable material properties. However, rational design of PEG hydrogel scaffold properties has been inhibited by the interdependence of key material properties such as modulus and mesh size. This study examined the effect of an acrylated 4-arm PEG cross-linker on gel modulus and mesh size as a means of inducing local increases in cross-link density to decouple these two parameters. It was determined that adding the 4-arm PEG cross-linker to PEG hydrogels resulted in statistically significant increases in both tensile and compressive modulus while having minimal effects on overall gel mesh size. The incorporation of the 4-arm PEG cross-linker also broadened the range of achievable mechanical properties. This study provides the methodology to independently tune PEG hydrogel modulus and mesh size, which may be utilized in future investigations of the individual and combined effects of PEG hydrogel modulus and mesh size on cell behavior and viability. It also presents a more finely tunable hydrogel scaffold with utility in a broad range of tissue engineering applications. PMID- 21626659 TI - Immobilization of Ag nanoparticles/FGF-2 on a modified titanium implant surface and improved human gingival fibroblasts behavior. AB - The objective of this study was to form a rapid and firm soft tissue sealing around dental implants that resists bacterial invasion. We present a novel approach to modify Ti surface by immobilizing Ag nanoparticles/FGF-2 compound bioactive factors onto a titania nanotubular surface. The titanium samples were anodized to form vertically organized TiO(2) nanotube arrays and Ag nanoparticles were electrodeposited onto the nanotubular surface, on which FGF-2 was immobilized with repeated lyophilization. A uniform distribution of Ag nanoparticles/FGF-2 was observed on the TiO(2) nanotubular surface. The L929 cell line was used for cytotoxicity assessment. Human gingival fibroblasts (HGFs) were cultured on the modified surface for cytocompatibility determination. The Ag/FGF 2 immobilized samples displayed excellent cytocompatibility, negligible cytotoxicity, and enhanced HGF functions such as cell attachment, proliferation, and ECM-related gene expression. The Ag nanoparticles also exhibit some bioactivity. In conclusion, this modified TiO(2) nanotubular surface has a large potential for use in dental implant abutment. PMID- 21626660 TI - Fibrillar films obtained from sodium soap fibers and polyelectrolyte multilayers. AB - An objective of tissue engineering is to create synthetic polymer scaffolds with a fibrillar microstructure similar to the extracellular matrix. Here, we present a novel method for creating polymer fibers using the layer-by-layer method and sacrificial templates composed of sodium soap fibers. Soap fibers were prepared from neutralized fatty acids using a sodium chloride crystal dissolution method. Polyelectrolyte multilayers (PEMs) of polystyrene sulfonate and polyallylamine hydrochloride were deposited onto the soap fibers, crosslinked with glutaraldehyde, and then the soap fibers were leached with warm water and ethanol. The morphology of the resulting PEM structures was a dense network of fibers surrounded by a nonfibrillar matrix. Microscopy revealed that the PEM fibers were solid structures, presumably composed of polyelectrolytes complexed with residual fatty acids. These fibrillar PEM films were found to support the attachment of human dermal fibroblasts. PMID- 21626661 TI - Arginine conjugation affects the endocytic pathways of chitosan/DNA nanoparticles. AB - Previous reports showed that arginine-rich peptides and oligoarginines facilitated the cellular internalization of DNA and proteins. A number of studies demonstrated that arginine-conjugated chitosan (CS)/DNA nanoparticles (ACGN) mediated significantly higher expression of the transgenes when compared with CS/DNA nanoparticles (CGN). However, the underlying mechanisms remain poorly understood. In the current study, the endocytic pathways through which cells internalize ACGN and CGN were explored by incubating the fluorescent CGN or ACGN with A10 cells in the presence of a variety of inhibitors of different endocytic pathways. The data accumulated in the current study revealed that conjugation of arginine moieties onto CS molecules enhanced the cellular uptake of the polymer/DNA nanoparticles and their transgenic efficacy, probably due to changes in the endocytic pathways, which led to the preference of internalization of ACGN by the caveolin-mediated endocytosis in comparison with that of CGN. These findings provide further support to the previous observations that ACGN mediated much higher expression of the transgenes when compared with CGN. PMID- 21626662 TI - Comparison of bacterial adhesion and cellular proliferation on newly developed three-dimensional scaffolds manufactured by rapid prototyping technology. AB - Scaffolds used in the field of tissue engineering should facilitate the adherence, spreading, and ingrowth of cells as well as prevent microbial adherence. For the first time, this study simultaneously deals with microbial and tissue cell adhesion to rapid prototyping-produced 3D-scaffolds. The cell growth of human osteosarcoma cells (CAL-72) over a time period of 3-11 days were examined on three scaffolds (PLGA, PLLA, PLLA-TCP) and compared to the adhesion of salivary microorganisms and representative germs of the oral flora (Porphyromonas gingivalis, Prevotella nigrescens, Candida albicans, Enterococcus faecalis, Streptococcus mutans, and Streptococcus sanguinis). Scanning electron microscopy (SEM), cell proliferation measurements, and determination of the colony forming units (CFU) were performed. The cell proliferation rates on PLLA and PLLA-TCP after 3, 7, and 11 days of cultivation were higher than on PLGA. On day 3 the proliferation rates on PLLA and PLLA-TCP, and on day 5 on PLLA-TCP, proved to be significantly higher compared to that of the control (culture plate). The strain which showed the most CFUs on all of the investigated scaffolds was P. gingivalis, followed by E. faecalis. No significant CFU differences were determined examining P. gingivalis among the biomaterials. In contrast, E. faecalis was significantly more adherent to PLGA and PLLA compared to PLLA-TCP. The lowest CFU values were seen with C. albicans and P. nigrescens. Salivary born aerobic and anaerobic microorganisms adhered significantly more to PLGA compared to PLLA-TCP. These results supported by SEM point out the high potential of PLLA-TCP in the field of tissue engineering. PMID- 21626663 TI - Preparation, characterization, in vitro bioactivity, and osteoblast adhesion of multi-level porous titania layer on titanium by two-step anodization treatment. AB - To combine the advantages of different electrolytes in anodic oxidation, pure titanium samples were anodized in CH(3) COOH electrolyte according to a novel anodizing treatment regime and then in H(2) SO(4) electrolyte in potentialstatic mode. The in vitro bioactivity of the as-prepared titanium samples was evaluated by simulated body fluid (SBF) test. In addition, MG63 osteoblast-like cells were cultured on surfaces of the as-prepared titanium samples to evaluate osteoblast adhesion ability. The titanium samples after the two-step anodization treatment were covered by titania layers of anatase and/or rutile with several micrometres thickness and presented a multi-level porous surface morphology consisting of interlaced grooves about 20-MUm wide overlaid with submicron scale pores. The SBF test results showed that the crystal titania layers prepared at appropriate conditions were able to induce apatite-forming in 7 days, indicating that the abundance of surface Ti-OH groups and (101)-oriented rutile structure both played important roles in in vitro bioactivity of titania layers. The cell experiment results showed that the macroscopic grooves could effectively promote osteoblast adhesion and growth and submicron scale pores might be beneficial to osteoblast adhesion. The two-step anodization treatment might be a promising candidate for surface modification of titanium implant. PMID- 21626664 TI - Modulation of immunological properties of allogeneic mesenchymal stem cells by collagen scaffolds in cartilage tissue engineering. AB - Influence of the structures of some collagen scaffolds on immunological properties of the seeded allogeneic mesenchymal stem cells (MSCs) was studied in this article. Hydrogels, sponge, and membrane were prepared from type-I collagen. These scaffolds were seeded with neonatal rabbit MSCs and cultured for different periods. Changes of the immunological properties associated with different scaffolds were analyzed and compared. It was found that the expression of major histocompatibility complex (MHC) class I and II molecules on MSCs increased gradually in all scaffolds, but the least increment was recorded in hydrogels. Mixed lymphocyte reactions (MLR) showed that the MSC-hydrogel constructs invoked considerably low allogeneic lymphocytes proliferation. Even in presence of interferon-gamma (IFN-gamma), the hydrogels with higher concentration gave comparatively lower increment of MHC-II expression and allogeneic lymphocytes proliferation. These results suggest that different scaffold structures may provide different microenvironments and extents of isolation from the host immune system for the seed cells, thereby affecting their immunological properties. Therefore, scaffold structures may modulate the immunological properties of tissue-engineered cartilage with allogeneic cells. Hydrogels, especially which were prepared from higher collagen concentrations, were found to be a promising scaffold structure, from the perspective of avoiding severe immune rejection. PMID- 21626665 TI - Covalent and ionic co-cross-linking--an original way to prepare chitosan-gelatin hydrogels for biomedical applications. AB - The first goal of this work was to develop a method for obtaining interpenetrating gelatin (G)-chitosan (CS) networks prepared by double cross linking (covalent followed by ionic) that exhibit hydrogel character. The second goal was to modulate their properties as a function of the preparation parameters by using neural network models. This study was therefore carried out by experiment and simulation. The covalent cross-linking resulted from the reaction between the carbonyl groups of glutaraldehyde with amino groups belonging to both polymers; the ionic cross-linking is based on the interaction between tripolyphosphate anions and protonated amine groups (ammonium ions) of the polymers. The total cross-linking density (indirectly assessed by estimating the water swelling capacity) and the ability to include hydrosoluble bioactive principles are influenced by the following process parameters: the CS/G ratio, the amount of ionic cross-linker, and the ionic cross-linking time. The prepared hydrogels were characterized with respect to their structural, morphological, and some physical properties. The hydrogels ability to load high amounts of water soluble drugs indicates their potential use as carriers for biologically active principles in the human body. A neural network methodology was applied to model the swelling degree and caffeine loading/release capacity depending on reaction conditions; in addition, applying this method, the optimal preparation conditions have been determined, targeting pre-established values for swelling degree or maximum caffeine value. The accuracy of the results obtained through this technique proves that the neural networks are suitable tools for modeling cross linking processes taking place complex nonlinear polymers. PMID- 21626666 TI - Understanding the impact of divalent cation substitution on hydroxyapatite: an in vitro multiparametric study on biocompatibility. AB - Hydroxyapatite (HA), a stable and biocompatible material for bone tissue therapy, may present a variable stoichiometry and accept a large number of cationic substitutions. Such substitutions may modify the chemical activity of HA surface, with possible impact on biocompatibility. In this work, we assessed the effects of calcium substitution with diverse divalent cations (Pb(2+), Sr(2+), Co(2+), Zn(2+), Fe(2+), Cu(2+), or Mg(2+)) on the biological behavior of HA. Physicochemical analyses revealed that apatite characteristics related to crystallinity and calcium dissolution/uptake rates are very sensitive to the nature of cationic substitution. Cytocompatibility was evaluated by mitochondrial activity, membrane integrity, cell density, proapoptotic potential, and adhesion tests. With the exception of Zn-HA, all the substituted HAs induced some level of apoptosis. The highest apoptosis levels were observed for Mg-HA and Co-HA. Cu-HA was the only material to impair simultaneously mitochondrial activity, membrane integrity, and cell density. The highest relative cell densities after exposure to the modified HAs were observed for Mg-HA and Zn-HA, while Co-HA significantly improved cell adhesion onto HA surface. These results show that changes on surface dissolution caused by cationic substitution, as well as the increase of metal species released to biological media, were the main responsible factors related to alterations on HA biocompatibility. PMID- 21626667 TI - A novel strontium-doped calcium polyphosphate/erythromycin/poly(vinyl alcohol) composite for bone tissue engineering. AB - It is our goal to develop bactericidal bone scaffolds with osteointegration potential. In this study, poly(vinyl alcohol) (PVA) coating (7%) was applied to an erythromycin (EM)-impregnated strontium-doped calcium polyphosphate (SCPP) scaffold using a simple slurry dipping method. MicroCT analysis showed that PVA coating reduced the average pore size and the percentage of pore interconnectivity to some extent. Compressive strength tests confirmed that the PVA coating significantly increased material elasticity and slightly enhanced the scaffold mechanical strength. It was also confirmed that the PVA coatings allowed a sustained EM release that is controlled by diffusion through the intact PVA hydrogel layer, irrespective of the drug solubility. PVA coating did not inhibit the EM bioactivity when the scaffolds were immersed in simulated body fluid for up to 4 weeks. EM released from SCPP-EM-PVA composite scaffolds maintained its capability of bacterial growth (S. aureus) inhibition. PVA coating is biocompatible and nontoxic to MC3T3 preosteoblast cells. Furthermore, we found that SCPP-EM-PVA composite scaffolds and their eluants remarkably inhibited RANKL induced osteoclastogenesis in a murine RAW 264.7 macrophage cell line. Thus, this unique multifunctional bioactive composite scaffold has the potential to provide controlled delivery of relevant drugs for bone tissue engineering. PMID- 21626668 TI - A patient with apparently reciprocal translocation and cryptic 10p deletion. PMID- 21626669 TI - Encephalocraniocutaneous lipomatosis (ECCL): neuroradiological findings in three patients and a new association with fibrous dysplasia. AB - Encephalocraniocutaneous lipomatosis (ECCL) is a rare neurocutaneous syndrome characterized by involvement of tissues of ectodermal and mesodermal origin such as skin, eye, adipose tissue, and brain. Since 1970, when Haberland and Perou had described the first patient, 54 cases of ECCL have been reported in literature. We report on three new boys with ECCL. In addition to their typical dermal, ocular and central nervous system anomalies, one of them had a spheno-ethmoidal osseous lesion. Histopathological evaluation confirmed the benign nature of the lesion and was consistent with fibrous dysplasia. The aim of our study is to review clinical records and brain imaging studies of these three new patients with ECCL and compare these findings with those reported in literature to better define the phenotypic spectrum and radiological findings in ECCL. PMID- 21626670 TI - De novo Xq11.11 microdeletion including ARHGEF9 in a boy with mental retardation, epilepsy, macrosomia, and dysmorphic features. AB - We report on a novel Xq11.11 microdeletion in a patient presenting with severe mental retardation (MR), focal epilepsy, tall stature, macrocephaly, and dysmorphism. This 1.3 Mb deletion, identified using array CGH, includes a single gene with known function-ARHGEF9-plus 1 gene with unknown function and three putative genes. ARHGEF9 encodes collybistin (Cb) that plays an important role in the localization of gephyrin which is the key protein of the scaffolding system of inhibitory synapses and is essential for postsynaptic clustering of both GABA(A) and glycine receptors. Cb-deficient male mice show reduced exploratory behavior, impaired spatial learning, increased anxiety scores, and reduction of gephyrin-dependent GABA receptor clusters in amygdala and hippocampus. Mutations or disruption of ARHGEF9 due to chromosomal rearrangements have been found in three patients with various clinical presentations: nevertheless, all 3 presented with MR and 2 with epilepsy. The case we report on provides further evidence for the role of ARHGEF9 in cognitive development. The other phenotypic features in our patient, including macrosomia and dysmorphism, may also be related to the loss of this gene. Alternatively, they may be consequences of the loss of one or more of the other genes located within the deletion or of the disruption of sequences regulating neighboring genes. Additional case reports with identical or overlapping deletions would help in defining the phenotype associated with ARHGEF9 haploinsufficiency. PMID- 21626671 TI - Prevalence of sleep problems in Smith-Lemli-Opitz syndrome. AB - Smith-Lemli-Opitz syndrome (SLOS) is an autosomal recessive genetic disorder, characterized by multiple congenital anomalies, and intellectual disability. It is caused by a genetically inherited deficiency of the enzyme 7 dehydrocholesterol-delta-7-reductase, which results in increased serum levels of 7-dehydrocholesterol (7-DHC), and decreased levels of cholesterol. This study assesses the prevalence of sleep problems in patients with SLOS. The study group comprised 18 subjects with SLOS, ages 2-31 years (median 10.7 +/- 8.5 years). Parents completed several questionnaires (Intake Demographic Form; Pediatric Sleep Questionnaire; Pediatric Daytime Sleepiness Scale). The SLOS subjects had symptoms of sleep-disordered breathing (50% snoring; 66.7% mouth breathing), problems with sleep onset [difficulty falling asleep (61.1%) sleep onset time >30 min (62%)], sleep maintenance [wake up screaming (61.1%), waking up more than twice (44.4%), having trouble falling back to sleep (66.7%), waking up early in the morning (61.1%), and restless sleep (50%)]. The subjects with SLOS needed parents in the room to fall asleep (50%), watch TV or listen to music to fall asleep (44.4%), and described bed sharing (33.3%), indicating sleep-anxiety and sleep-associations. Symptoms of excessive-daytime-sleepiness were frequently reported [un-refreshed in the morning (38.9%), daytime sleepiness (44.4%), and daytime naps (55.6%)]. Parents frequently observed difficulty of organizing tasks (66.7%), and easy distractibility (88.9%). Sleep problems such as sleep disordered breathing, sleep-related anxiety and sleep associations, disturbed sleep patterns at night, and excessive daytime sleepiness are frequent in children with SLOS. PMID- 21626672 TI - Fanconi anemia-like presentation in an infant with constitutional deletion of 21q including the RUNX1 gene. AB - We describe a newborn female with a de novo interstitial deletion of chromosome 21q21.1-22.12 including the RUNX1 gene who had developmental delay, multiple congenital anomalies, tetralogy of Fallot, anemia, and chronic thromobocytopenia requiring frequent platelet transfusions from birth. Because of her physical and hematologic abnormalities, she was tested for Fanconi anemia (FA). Lymphocytes and fibroblasts from this patient demonstrated increased chromosome breakage with exposure to the clastogen mitomycin C, but not, in contrast to most FA patients, to diepoxybutane. Further testing by Western analysis and complementation testing did not show a defect in the function of known Fanconi proteins. Her constitutional deletion was later found to span 13.2 Mb by chromosome microarray analysis, encompassing the RUNX1 gene that has been implicated in thrombocytopenia and predisposition to acute myelogenous leukemia (AML) when in the haploinsufficient state. We compare her phenotype to other individuals with similar 21q deletions and thrombocytopenia, as well as those with FA. We suggest that deletion of RUNX1 or another critical gene within the deleted region may result in chromosomal instability similar to that seen in FA. PMID- 21626673 TI - Rett syndrome: a study of the face. AB - Rett syndrome is a unique disorder of neurodevelopment that is characterized by an evolving behavioral and developmental phenotype, which emerges after an apparently normal early infantile period. It almost exclusively affects females. The face of Rett syndrome is said to resemble that of Angelman syndrome, although there seems little objective support for this impression and it is not a concept with universal support. This observational and anthropometric study was carried out to define the key facial characteristics of females with Rett syndrome and to evaluate whether any changes of significance occur with age. Thirty-seven affected Caucasian females, from 2 to 20 years of age, were evaluated. Thirty five of them had a documented mutation in MECP2 while the remaining two fulfilled the clinical criteria for Rett syndrome and had been diagnosed by an experienced clinician. Few unusual facial features were noted. Almost all facial measurements were within the normal range although head circumference tended to fall below the normal range with increasing age. The pattern of measurements was constant over time, with the exception of increased facial width in the under 3-year-old girls. The face of Rett syndrome does not demonstrate marked prognathism, wide mouth, spaced teeth or striking microcephaly, all features of Angelman syndrome. Thus, while Rett and Angelman syndromes have similar clinical, neurological, and behavioral phenotypes, they do not appear to share similar facial features. PMID- 21626674 TI - Submicroscopic deletion in 7q31 encompassing CADPS2 and TSPAN12 in a child with autism spectrum disorder and PHPV. AB - We performed array comparative genomic hybridization utilizing a whole genome oligonucleotide microarray in a patient with the autism spectrum disorders (ASDs) and persistent hyperplastic primary vitreous (PHPV). Submicroscopic deletions in 7q31 encompassing CADPS2 (Ca(2+) -dependent activator protein for secretion 2) and TSPAN12 (one of the members of the tetraspanin superfamily) were confirmed. The CADPS2 plays important roles in the release of neurotrophin-3 and brain derived neurotrophic factor. Mutations in TSPAN12 are a relatively frequent cause of familial exudative vitreoretinopathy. We speculate that haploinsufficiency of CADPS2 and TSPAN12 contributes to ASDs and PHPV, respectively. PMID- 21626675 TI - Expanding the phenotype of gingival fibromatosis-mental retardation hypertrichosis (Zimmermann-Laband) syndrome. AB - Zimmermann-Laband syndrome (ZLS) is a rare disorder characterized by gingival fibromatosis, hypertrichosis, intellectual disability, and absence and/or hypoplasia of the nails or terminal phalanges of the hands and feet. The syndromic features of ZLS are highly variable and can overlap with other entities featuring gingival fibrosis. This study describes a patient with ZLS with novel findings, including colpocephaly, hemivertebra, polydactyly, hyperpigmentation, and hemihyperplasia. Thus, the present report expands the phenotypic spectrum of this uncommon syndrome. PMID- 21626676 TI - Array-CGH study of partial trisomy 9p without mental retardation. AB - Partial trisomy 9p is one of the most common detected autosomal structural anomalies, so the phenotype-genotype correlation of this rearrangement has been well described. Despite variation in size of the 9p duplications, trisomy 9p syndrome is characterized by typical dysmorphic features and a variable but constant psychomotor and mental retardation. Previously reported phenotype genotype correlation studies proposed that the critical region for phenotype is located in 9p22. We report here on a new patient with partial trisomy 9p13.3 >9pter in an 8-year-old boy with typical trisomy 9p dysmorphic features but a normal mental development. Cytogenetics investigations showed that our patient karyotype was 47,XY,+ der(22)t(9;22)(p13.q11) inherited by a 3:1 disjunction of a maternal reciprocal translocation t(9;22)(p13.q11). FISH and array CGH analysis were used to better characterize duplicated chromosomal regions and showed a large duplication of chromosome 9p13.3->9pter associated to microduplication in 22q11.1. The size of the duplications in chromosomes 9p and 22q were estimated about 33.9 and 2.67 Mb, respectively. The comparison between this case and those reported in the literature allows us to support that all syndromes show variability and that not all partial trisomies 9p are associated with intellectual disability. PMID- 21626677 TI - Isolated oligodontia associated with mutations in EDARADD, AXIN2, MSX1, and PAX9 genes. AB - Oligodontia is defined as the congenital lack of six or more permanent teeth, excluding third molars. Oligodontia as well as hypodontia (lack of one or more permanent teeth) are highly heritable conditions associated with mutations in the AXIN2, MSX1, PAX9, EDA, and EDAR genes. Here we define the prevalence of mutations in the AXIN2, MSX1, PAX9, EDA, and EDAR genes, and the novel candidate gene EDARADD in a cohort of 93 Swedish probands with non-syndromic, isolated oligodontia. Mutation screening was performed using denaturing gradient gel electrophoresis and DNA sequence analysis. Analyses of the coding sequences of the six genes showed sequence alterations predicted to be damaging or potentially damaging in ten of 93 probands (10.8%). Mutations were identified in the EDARADD (n = 1), AXIN2 (n = 3), MSX1 (n = 2), and PAX9 (n = 4) genes, respectively. None of the 10 probands with mutations had other self-reported symptoms from ectodermal tissues. The oral parameters were similar when comparing individuals with and without mutations but a family history of oligodontia was three times more frequent for probands with mutations. EDARADD mutations have previously been reported in a few families segregating hypohidrotic ectodermal dysplasia and this is, to our knowledge, the first report of an EDARADD mutation associated with isolated oligodontia. PMID- 21626678 TI - 5q14.3 neurocutaneous syndrome: a novel continguous gene syndrome caused by simultaneous deletion of RASA1 and MEF2C. AB - Haploinsufficiency of RASA1, located on chromosome 5q14.3, has been identified as the etiology underlying the disorder capillary malformation-arteriovenous malformation (CM-AVM). Recently, haploinsufficiency of MEF2C, located 1.33 Mb distal to RASA1 on chromosome 5q14.3, has been implicated as the genetic etiology underlying a complex array of deficits including mental retardation, hypotonia, absent speech, seizures, and brain anomalies. Here we report a patient who is haploinsufficient in both RASA1 and MEF2C who presents with dermatologic and neurologic abnormalities that constitute a 5q14.3 neurocutaneous syndrome. This finding highlights the need to assess for CM-AVM in patients with neurologic features consistent with MEF2C haploinsufficiency, and vice versa. PMID- 21626679 TI - The 3q29 microdeletion syndrome: report of three new unrelated patients and in silico "RNA binding" analysis of the 3q29 region. AB - The human 3q29 microdeletion syndrome is associated with mild facial dysmorphism, developmental delay and variable congenital malformations. We report three new unrelated patients with this syndrome. We also performed in silico RNA binding analysis in silico on the 3q29 critical region genes. Several genes within this genomic region including DLG1 and RNF168 are predicted to bind RNA. While recessive mutations in RNF168 cause RIDDLE syndrome, an immune deficiency and radiosensitivity disorder, the potential impact of heterozygous deletion of RNF168 on patients with the 3q29 deletion syndrome is still unknown. PMID- 21626680 TI - A 10.46 Mb 12p11.1-12.1 interstitial deletion coincident with a 0.19 Mb NRXN1 deletion detected by array CGH in a girl with scoliosis and autism. AB - We present a 12-year-old girl with de novo karyotype 46,XX,del(12)(p11.1p12.1). Array CGH revealed in addition to a 10.466 Mb interstitial deletion on 12p11.1 >12p12.1 a 0.191 Mb deletion on 2p16.3. The girl presented with mild facial dysmorphism consisting of microcephaly, hypertelorism, downslanting palpebral fissures, strabismus, broad nasal base, bulbous nose, short philtrum, micro/retrognathia, irregular tooth arrangement, phalangeal deformity in distal phalanges of hands, 5th finger camptodactyly, brachydactyly in feet, history of joint hypermobility, and scoliosis. She was considered to have mild to moderate mental retardation and ascertained for an autism spectrum disorder(ASD). Short arm of chromosome 12 interstitial deletions are rarely reported whereas point mutations and deletions of NRXN1, which is located on chromosome 2p16.3, are associated with ASDs. In this article we present and discuss the phenotypic consequences of a patient who was affected by deletions of two different chromosomal regions. PMID- 21626682 TI - Functional hydrogel density patterns fabricated by dip-pen nanolithography and photografting. PMID- 21626683 TI - Multicomponent DNA-templated nanoparticle chains with controllable dimension and composition. PMID- 21626684 TI - Separation of chemical reaction intermediates by metal-organic frameworks. AB - HPLC columns custom-packed with metal-organic framework (MOF) materials are used for the separation of four small intermediates and byproducts found in the commercial synthesis of an important active pharmaceutical ingredient in methanol. In particular, two closely related amines can be separated in the methanol reaction medium using MOFs, but not with traditional C18 columns using an optimized aqueous mobile phase. Infrared spectroscopy, UV-vis spectroscopy, X ray diffraction, and thermogravimetric analysis are used in combination with molecular dynamic simulations to study the separation mechanism for the best performing MOF materials. It is found that separation with ZIF-8 is the result of an interplay between the thermodynamic driving force for solute adsorption within the framework pores and the kinetics of solute diffusion into the material pores, while the separation with Basolite F300 is achieved because of the specific interactions between the solutes and Fe(3+) sites. This work, and the exceptional ability to tailor the porous properties of MOF materials, points to prospects for using MOF materials for the continuous separation and synthesis of pharmaceutical compounds. PMID- 21626685 TI - Hybrid nanomotor: a catalytically/magnetically powered adaptive nanowire swimmer. AB - A synthetic hybrid nanomotor, which combines chemically powered propulsion and magnetically driven locomotion, is described. The new catalytic-magnetic nanomotor consists of a flexible multisegment Pt-Au-Ag(flex)-Ni nanowire, with the Pt-Au and Au-Ag(flex)-Ni portions responsible for the catalytic and magnetic propulsion modes, respectively. The experimental data and theoretical considerations indicate that the hybrid design only minimally compromises the individual propulsion modes. Rapid and convenient switching from the catalytic to the magnetic mode is illustrated. The resulting catalytic-magnetic adaptive nanomotor can address the fuel depletion and salt limitation common to chemically powered motors by switching to magnetic propulsion. Reversal of the motion direction is also achieved upon applying the magnetic field. Such use of two sources to power a hybrid device offers a broader scope of operation and holds considerable promise for designing adaptive nanovehicles that reconfigure their operation in response to environmental changes or unexpected events. PMID- 21626686 TI - Synthesis of macroporous calcium carbonate/magnetite nanocomposites and their application in photocatalytic water splitting. PMID- 21626687 TI - Using charge to control the functional properties of self-assembled nanopores in membranes. PMID- 21626688 TI - Quantification of uptake and localization of bovine serum albumin-stabilized single-wall carbon nanotubes in different human cell types. AB - Single-wall carbon nanotubes (SWCNTs) possess many unique, inherent properties that make them attractive materials for application in medical and biological technologies. Development of concentrated SWCNT dispersions of isolated nanotubes that retain SWCNTs' inherent properties with minimal negative cellular effects is essential to fully realize the potential of SWCNTs in biotechnology. It is shown that bovine serum albumin (BSA), a common and well-characterized model blood serum protein, can individually disperse SWCNTs at concentrations of up to 0.3 mg mL(-1) while retaining SWCNTs' optical properties. Uptake into human mesenchymal stem cells (hMSC) and HeLa cells is quantified, revealing strikingly high concentrations of 86 +/- 33 * 10(6) and 21 +/- 13 * 10(6) SWCNTs per cell, respectively, without any apparent acute deleterious cellular effects. Through high-resolution confocal Raman spectroscopy and imaging, it is established that SWCNT-BSAs are preferentially localized intracellularly, especially in the cytoplasm of both hMSCs and HeLa cells. The uptake and localization results demonstrate the efficacy of BSA as a biocompatible dispersant and a mediator of bioactivity. BSA is widely available and inexpensive, which make these concentrated, highly-dispersed, noncovalently modified SWCNT-BSAs suitable for the development of SWCNT-based biotechnologies. PMID- 21626689 TI - Self-aligned sub-10-nm nanogap electrode array for large-scale integration. AB - A novel approach to creating a gap on the nanometer scale between two adjacent electrodes of the same or different metals is described. The gap size can be well controlled through sidewall coverage in a self-aligned manner and it can be tuned from 60 nm down to 5 nm with high reproducibility. This technique is fully compatible with traditional microfabrication technology and it is easily implemented to fabricate a nanogap electrode array for integration purposes. An array of short-channel single-walled carbon nanotube field-effect transistors is demonstrated. PMID- 21626690 TI - Flexible ZnO-cellulose nanocomposite for multisource energy conversion. AB - Materials with the ability to harness multiple sources of energy from the ambient environment could lead to new types of energy-harvesting systems. It is demonstrated that nanocomposite films consisting of zinc oxide nanostructures embedded in a common paper matrix can be directly used as energy-conversion devices to transform mechanical and thermal energies to electric power. These mechanically robust and flexible devices can be fabricated over large areas and are capable of producing an output voltage and power up to 80 mV and 50 nW cm(-2) , respectively. Furthermore, it is shown that by integrating a certain number of devices (in series and parallel) the output voltage and the concomitant output power can be significantly increased. Also, the output voltage and power can be enhanced by scaling the size of the device. This multisource energy-harvesting system based on ZnO nanostructures embedded in a flexible paper matrix provides a simplified and cost-effective platform for capturing trace amounts of energy for practical applications. PMID- 21626691 TI - Design and application of inorganic nanoparticle superstructures: current status and future challenges. AB - Self-assembly of inorganic nanoparticles (NPs) into superstructures, which is used as a general way to integrate functional inorganic NPs into macroscale devices, has attracted much research interest. This review will summarize the recent progress and discuss future challenges of the inorganic NP superstructures. Examples include both DNA-based and polymer-based NP assemblies with controlled positioning and geometries, and quasicrystalline ordered structures from the self-assembly of binary or ternary NPs. Different from their individual NP counterparts, these self-assembled superstructures possess unique properties, such as optical chirality and dynamic structural change under an external stimulus. Due to their diversified structures and functionalities, inorganic NP superstructures have shown a wide range of promise for applications in electronic and photonic devices, such as field-effect transistors, magnetoresistive components, optical information recording, and solar cells. PMID- 21626692 TI - Exploiting dimerization of purely peptidic amphiphiles to form vesicles. PMID- 21626693 TI - Ultrasonic nebulization extraction coupled with headspace hollow fiber microextraction of pesticides from root of Panax ginseng C.A. Mey. AB - The ultrasonic nebulization extraction coupled with headspace hollow fiber microextraction (UNE-HS-HFME) was applied for the extraction of pesticides from root of Panax ginseng C.A. Mey. Experimental parameters, which affect the performances of ultrasonic nebulization extraction coupled with headspace hollow fiber microextraction, such as the kind of acceptor solvent in the pore of the fiber wall, the sample amount, extraction time, salt concentration in extraction solvent, pH of the acceptor solution, the elution time, and times were studied and optimized. The analytes were determined by high-performance liquid chromatography. The detection limits for simeton, monolinuron, chlortoluron, karmex, and prebane are 20.9, 18.4, 18.2, 12.4, and 22.2 MUg/kg, respectively. Besides volatile and semi-volatile compounds, the non-volatile compounds also can be determined by the proposed method. The extraction and enrichment process can be performed simultaneously. PMID- 21626694 TI - Stability evaluation of tramadol enantiomers using a chiral stability-indicating capillary electrophoresis method and its application to pharmaceutical analysis. AB - In this study, a chiral stability-indicating CE assay was developed for the stability evaluation of tramadol (TR) enantiomers in commercial tablets using maltodextrin as chiral selector. To investigate the stability-indicating power of the analytical method as well as stability evaluation of TR enantiomers, active pharmaceutical ingredient and TR tablets were subjected to photolysis, heat, oxidation and hydrolysis to conduct stress testing. Best separation for the TR enantiomers was achieved on an uncoated fused-silica capillary at 20 degrees C using borate buffer (50 mM, pH 10.2) containing 10% m/v maltodextrin. All determinations were performed by a UV detector at 214 nm. A constant voltage of 20 kV was applied to obtain the separation. The range of quantitation for both enantiomers was 5-100 MUg/mL (R>0.996). Intra- and inter-day RSD (n=6) were less than 10%. The percent relevant errors were obtained to be less than 4.0 for both enantiomers. The limits of quantitation and detection for both enantiomers were 5 and 1.5 MUg/mL, respectively. Degradation products resulting from the stress studies were the same for both enantiomers and did not interfere with the detection of the enantiomers. PMID- 21626695 TI - Modeling of retention behaviors of most frequent components of essential oils in polar and non-polar stationary phases. AB - The gas chromatography retention indices of 100 different components of essential oils, on three columns with stationary phases of different polarities, were used to develop robust quantitative structure-retention relationship (QSRR) models. Two linear models with only one variable, i.e. solvation entropy, were developed, which explain 95 and 94% of variances of the test set for dimethyl silicone and dimethyl silicone with 5% phenyl group columns, respectively. These models are extremely simple and easy to interpret, but they show higher errors compared with more robust models such as partial least square (PLS) and ridge regressions. For the third column (polyethylene glycol (PEG)), 24 hydrogen bonding descriptors were calculated and were used for modeling. Kernel orthogonal projection to latent structure (KOPLS), as a non-linear technique, was applied for the modeling of the retention indices of the compounds on the PEG column. R(2) values for the test set established by Monte Carlo cross-validation and SPXY (sample set partitioning based on joint x-y distances) test set of the KOPLS were 0.92 and 0.94, respectively. y-Randomization indicated that chance plays no role in constructing the KOPLS model. PMID- 21626696 TI - Phenylaminopropyl-functionalized stationary phase for open-tubular capillary electrochromatography of alkaloids and aromatic acids. AB - A multi-functional open-tubular (OT) column covalently modified with [3 (phenylamino)propyl]trimethoxysilane (PPTMS) via a single-step silanization reaction has been developed, and employed for alkaloids and aromatic acids analytes by CEC. Both anodic and cathodic EOF could be gained in the PPTMS-bonded column, and anodic EOF was exhibited when the pH of running buffer was less than 8.0. Using thiourea as the EOF marker, a satisfactory column stability was denoted, and the RSD values for migration time and peak area were gained at 0.5 and 3.7% (intra-day, n=5), 1.7 and 5.6% (inter-day, n=3), 3.9 and 5.8% (column-to column, n=16 in four batches). With anodic EOF mode (pH=2.0 of 10 mmol/L phosphate buffer), favorable separations of cationic alkaloids (viz. uridine, adenosine, cytidine, adenine, cytosine) were successfully achieved with column efficiencies ranging from 95,000 to 187,000 plates/m, and the undesired adsorption on the inter-wall of capillary column could be avoided. Besides, six anionic aromatic acids could be also separated efficiently in the co electroosmotic mode with anodic EOF. And high efficiencies ranged from 176,000 to 235,000 plates/m were gained with a good repeatability. PPTMS-bonded capillary column might be used as an alternative functional medium to physical coating capillary column for the analyses of aromatic organic acids and bases. PMID- 21626697 TI - Optimized mobile phase for CEC of acetylsalicylic acid and its impurities using a methacrylate-based monolithic column. AB - In the past few years, monolithic methacrylate-based columns have attracted some attention in separation science. The mobile-phase optimization on these columns for drug analysis has not yet been thoroughly examined. This paper evaluates the separation of acetylsalicylic acid and its impurities as a case study. First, the best pH was determined as 2.3. Methacrylate-based phases can be employed at such pH because they remain charged, necessary to generate electro-osmotic flow. Then, a suitable solvent strength was determined. Trifluoroacetic acid (0.1%) was added to the mobile phase to improve peak shapes. The optimal organic modifier composition was then determined, using isoeluotropic mobile phases, based on the theory of Snyder's solvent triangle. Quadratic models were built to predict the retention of the compounds at all mobile-phase compositions within the triangle. The predictions were tested and found appropriate. Eventually, a baseline separation of acetylsalicylic acid and its impurities was not obtained. However, it could be concluded that one can optimize the mobile phase on methacrylate based monolithic columns in CEC using Snyder's solvent triangle approach. PMID- 21626698 TI - Improved chromatographic performances of glycidyl methacrylate anion-exchange monolith for fast nano-ion exchange chromatography. AB - An efficient and reproducible photopolymerized poly(glycidyl methacrylate-co ethylene dimethacrylate) was synthesized in Teflon-coated fused-silica capillaries (100 MUm id) and functionalized by reaction of triethylamine with reactive epoxy groups. We report here the successful transfer of a standard polymerization mixture optimized for the thermally initiated synthesis of glycidyl-based monolith to photo-induced polymerization. The monolith obtained after optimization of the photo-initiation conditions was characterized in reverse-phase chromatography evaluating its suitability in terms of efficiency, retention and hydrodynamic permeability. Reproducibility of the photo-induced procedure was satisfactory with RSD below 6% for retention and efficiency and slightly higher for hydrodynamic permeability (12%). The functionalized generic support was then used in nano-ion-exchange chromatography. Efficiencies up to 75,000 plates/m, ion-exchange capacity of 8 nano-equivalents/cm of monolithic column, with a combination of a satisfactory hydrodynamic permeability allowed to perform fast separations of five inorganic anions in <3 min maintaining baseline resolution. The efficiency of the monolith was not retention-dependent, demonstrating its wide range of possible applications for highly retained anions. PMID- 21626699 TI - Discovery, structure-activity relationship studies, and crystal structure of nonpeptide inhibitors bound to the Shank3 PDZ domain. AB - Shank is the central scaffolding protein of the postsynaptic density (PSD) protein complex found in cells of the central nervous system. Cellular studies indicate a prominent role of the protein in the organization of the PSD, in the development of neuronal morphology, in neuronal signaling, and in synaptic plasticity, thus linking Shank functions to the molecular basis of learning and memory. Mutations in the Shank gene have been found in several neuronal disorders including mental retardation, typical autism, and Asperger syndrome. Shank is linked to the PSD complex via its PDZ domain that binds to the C-terminus of guanylate-kinase-associated protein (GKAP). Here, small-molecule inhibitors of Shank3 PDZ domain are developed. A fluorescence polarization assay based on an identified high-affinity peptide is established, and tetrahydroquinoline carboxylates are identified as inhibitors of this protein-protein interaction. Chemical synthesis via a hetero-Diels-Alder strategy is employed for hit optimization, and structure-activity relationship studies are performed. Best hits possess K(i) values in the 10 MUM range, and binding to the PDZ domain is confirmed by 1H,15N HSQC NMR experiments. One of the hits crystallizes with the Shank3 PDZ domain. The structure, analyzed at a resolution of 1.85 A, reveals details of the binding mode. Finally, binding to PDZ domains of PSD-95, syntrophin, and DVL3 was studied using 1H,15N HSQC NMR spectroscopy. PMID- 21626703 TI - On the role of flexibility in protein-ligand interactions: the example of p53 tetramerization domain. AB - The recognition of protein surfaces by designed ligands has become an attractive approach in drug discovery. However, the variable nature and irregular behavior of protein surfaces defy this new area of research. The easy to understand "lock and-key" model is far from being the ideal paradigm in biomolecular interactions and, hence, any new finding on how proteins and ligands behave in recognition events paves a step of the way. Herein, we illustrate a clear example on how an increase in flexibility of both protein and ligand can result in an increase in the stability of the macromolecular complex. The biophysical study of the interaction between a designed flexible tetraguanidinium-calix[4]arene and the tetramerization domain of protein p53 (p53TD) and its natural mutant p53TD-R337H shows how the floppy mutant domain interacts more tightly with the ligand than the well-packed wild-type protein. Moreover, the flexible calixarene ligand interacts with higher affinity to both wild-type and mutated protein domains than a conformationally rigid calixarene analog previously reported. These findings underscore the crucial role of flexibility in molecular recognition processes, for both small ligands and large biomolecular surfaces. PMID- 21626704 TI - Encapsulation of pyrene-functionalized poly(benzyl ether) dendrons into a water soluble organometallic cage. AB - Two generations of lipophilic pyrenyl functionalized poly(benzyl ether) dendrimers (P(1) and P(2)) have been synthesized. The thermal properties of the two functionalized dendrimers have been investigated, and the pyrenyl group of the dendritic molecules encapsulated in the arene-ruthenium metalla-cage, [Ru(6)(p-cymene)(6)(tpt)(2)(donq)(3)](6+) ([1](6+)) (tpt=2,4,6-tri(pyridin-4-yl) 1,3,5-triazine; donq=5,8-dioxydo-1,4-naphthoquinonato). The host-guest properties of [P(1)?1](6+) and [P(2)?1](6+) were studied in solution by NMR and UV/Vis spectroscopic methods, thus allowing the determination of the affinity constants. Moreover, the cytotoxicity of these water-soluble host-guest systems was evaluated on human ovarian cancer cells. PMID- 21626706 TI - Dealing with crystal forms (the Kingdom of Serendip?). AB - The variety of crystal forms that may be associated with one specific molecule of interest can be extremely large: in addition to polymorphs, all sorts of crystalline solids can be obtained, from molecular and ionic co-crystals to hydrates/solvates, to, needless to say, polymorphs of all these new crystal forms. Lack of predictability of crystallization experiments, far from representing a failure or a nuisance, should encourage in the crystal maker the same attitude shown by the three princes of Serendip (who were making unexpected discoveries by virtue of their "sagacity and readiness of mind") to be ready to pick new avenues as the crystal experiments will yield something unplanned for. PMID- 21626707 TI - Thermally stable intermolecular proton bonds in polyaromatic aldehyde crystals. AB - We have synthesized self-assembled red-colored proton complexes of the aldehyde derivatives of polyaromatic hydrocarbon with strong intermolecular hydrogen bonding. These intermolecularly proton-bonded polyaromatic aldehydes formed as 1 pyrenecarbaldehyde (Py-CHO) reacted with HAuCl(4) to produce [(Py CHO)(2)H][AuCl(4)] under dry conditions. The formation of [(Py-CHO)(2)H][AuCl(4)] was confirmed by single-crystal structure determination and IR spectral analysis at various temperatures. The compounds are distinctively hydrophobic in nature and are soluble only in a few organic polar solvents. The proton bonds are clearly observed from both the electron density in X-ray analysis and the characteristic IR frequency signature. The proton complex units have an O-H(+)-O distance of the typical Zundel-like cationic hydrogen bond (in which two O atoms share a proton-like H in the midpoint of the short O-O distance of ~2.4 A). The proton bonds are thermally stable, even over 100 degrees C, because the complexes are stabilized in layered structures with pi-pi intermolecular interactions of the polyaromatic hydrocarbon ligands. The IR signatures at around 900, 1200, and 1700 cm(-1) for the Zundel-like proton bond are clearly characterized. PMID- 21626708 TI - Gold(I)-catalyzed cyclization of alkynyl hydrogen phosphates. PMID- 21626709 TI - Synthesis of unsymmetrical arylheteroarylmethanes by direct "on water" cross coupling between benzylic and heteroaromatic halides. PMID- 21626710 TI - Transesterification to biodiesel with superhydrophobic porous solid base catalysts. PMID- 21626711 TI - Increased incidence of bronchopulmonary fistulas complicating pediatric pneumonia. AB - BACKGROUND: The frequency of complicated pneumococcal disease, including necrotizing pneumonia, has increased over the last decade. During 2008-2009, we noted an increase in the number of children whose empyema was complicated by the development of a bronchopleural fistula and air leak. We studied these children to see if there was an associated cause. METHODS: This was a retrospective case note and database review of children admitted to our tertiary unit with a parapneumonic effusion or empyema from 2002 to 2007, compared with 2008 to 2009. For the latter period, we also compared the outcomes of those with a bronchopleural fistula to those without. RESULTS: During the 8-year period, 310 children were admitted. In the first 6 years, the frequency of air leaks was 1% (2/258) rising to 33% (16/49) in the last 2 years (P<0.0001). Three children were excluded as their fistulas were possibly iatrogenic. This was associated with a significant increase in median hospital stay (7 vs. 10 days, P<0.0001) and surgical intervention rate (2% vs. 14%, P=0.001). In the latter 2 years, S. pneumoniae serotype 3 was identified in 10/16 (91%) of those with a bronchopleural fistula compared to 1/33 (3%) of those without. CONCLUSIONS: The frequency of bronchopleural fistulas increased markedly in the 2 years 2008-2009. Although these cases were associated with pneumococcal serotype 3 infection, which was not covered by the heptavalent pneumococcal vaccine Prevenar(r) in use at that time, we do not know whether the increased incidence of fistulas was due to a change in serotype 3 prevalence. PMID- 21626712 TI - Flexible bronchoscopy as a valuable diagnostic and therapeutic tool in pediatric intensive care patients: a report on 5 years of experience. AB - OBJECTIVE: To evaluate the clinical role of flexible bronchoscopy (FB) in pediatric and neonatal intensive care units (ICUs). DESIGN: A retrospective review of all patients receiving FB procedures between January 2005 and December 2009. SETTING: Pediatric and neonatal ICUs of a tertiary care multi-disciplinary teaching hospital located in northern Taiwan. PATIENTS: A total of 358 ICU patients (223 males) who received 725 FB procedures. MEASUREMENTS AND MAIN RESULTS: The medical records were reviewed and analyzed. Mean age for the first time FB was 35.7 (+/-48.9 SD) months old and 68.2% of them were <3 years old. Among them, unexplained retraction or tachypnea (32.0%) and stridor (20.1%) were the two leading indications for FB. The positive finding rate of FB was 87.2%, with airway malacia (47.8%) being the most common, especially in patients <3 years old. Positive lesion sites were approximately equally distributed between the upper (51.1%) and lower (50.6%) airways. Concomitant findings in the esophagus were found in 15.4% of the patients. There were 518 interventional FBs (71.4%, 518/725 procedures) which were performed on 201 (56.1%, 201/358) patients; FB-aided endotracheal intubation (180/518, 34.7%) and laser therapy (109/518, 21.0%) were the two leading techniques used. No patient suffered from any long-term complications or mortality associated with the FB procedures. CONCLUSIONS: FB is a safe and valuable diagnostic and therapeutic tool for patients in pediatric and neonatal ICUs. PMID- 21626713 TI - Body satisfaction, nutritional adherence, and quality of life in youth with cystic fibrosis. AB - Children with cystic fibrosis (CF) typically have similar nutritional intake as healthy peers, despite recommendations to consume more calories and fat. Body satisfaction may play a role in nutritional intake: females may be content with their smaller size despite recommendations for weight gain, while males may desire to be larger and more muscular, which is more congruent with medical advice. Females are especially at risk, given their propensity to desire a smaller body size, tendency for lower HRQOL, steeper trajectory of health decline and shorter life expectancy than males. This study evaluated body satisfaction in relation to nutritional adherence and HRQOL in youth with CF. Fifty-four individuals with CF (age 9-17) completed the Cystic Fibrosis Questionnaire Revised (CFQ-R), the Figure Rating Scale, and a 24-hr diet recall interview with their caregiver. Twenty-four percent of youth were non-adherent with caloric goals, and 40.7% did not obtain the minimum recommendation for fat intake. Youth were classified as inconsistent with treatment goals (TI) if they desired a smaller body size or were content with their current size despite a BMI less than the 50th percentile; 44.8% of females were classified as TI, compared to only 8% of males. Statistical analyses were performed to evaluate the impact of gender and body satisfaction on HRQOL in youth with CF. Linear multiple regression models were fit; TI females had Emotional HRQOL scores 23 points lower than males. Results suggest that improving body satisfaction, especially for females, may help to improve overall quality of life and potentially impact nutritional adherence. PMID- 21626714 TI - Exhaled breath temperature in healthy children is influenced by room temperature and lung volume. AB - BACKGROUND: Exhaled breath temperature (EBT) has been proposed for the non invasive assessment of airway inflammation. Previous studies have not examined the influence of room temperature or lung size on the EBT. OBJECTIVE: This study aimed to address these issues in healthy children. METHODS: We assessed the effects of room temperature and lung volume in 60 healthy children aged 9-11 years (mean age 10.3 years, 33 male). Static lung volumes were assessed using multiple breath nitrogen washout. Questionnaire and skin prick tests were also used to establish respiratory health in the children. We obtained the EBT parameters of slope, end plateau temperature (PLET) and normalized plateau temperature (nPLET; plateau temperature minus inspired air temperature), and ascertained physiological factors influencing EBT. RESULTS: End plateau temperature was shown to be proportionally affected by room temperature (r = 0.532, P < 0.001) whereas slope and nPLET decreased with increasing room temperature (r = -0.392 P < 0.02 and r = -0.507 P = 0.002). After adjusting for room temperature, height and age, the total lung capacity (r(2) = 0.435, P = 0.006) and slow vital capacity (SVC; r(2) = 0.44, P = 0.005) were found to be the strongest predictors of end PLET in healthy children. When all factors were included in a multiple regression model, SVC and room temperature were the only predictors of plateau and nPLET. Slope was only influenced by room temperature. CONCLUSIONS: Exhaled breath temperature measurements are highly feasible in children with a 95% success rate in this healthy population. Room temperature and SVC significantly influence EBT variables in healthy children. Further studies are required to investigate the ability of EBT to assess airway inflammation in children with respiratory disease. Pediatr. Pulmonol. 2011; 46:1062-1068. (c) 2011 Wiley Periodicals, Inc. PMID- 21626715 TI - Analysis of 170 cases of congenital TB reported in the literature between 1946 and 2009. AB - INTRODUCTION: Congenital tuberculosis is a rare disease. The mortality is very high. Through a review of our own cases and the world literature, we describe clinical manifestations, treatment, and prognosis of this disease. METHODS: A total of 170 subjects with congenital tuberculosis that 6 cases identified by the authors and 164 cases identified in other case series were included in this study. All patients were diagnosed according to Cantwell's criteria. The data were analyzed using SPSS, version 17.0 spss. RESULTS: There were 70 premature babies among the 170 infants with congenital tuberculosis. The average onset age was 20 days. The mothers of 162 patients were diagnosed as having active tuberculosis during pregnancy or after parturition. Nonspecific signs and symptoms were found in these 170 cases, such as fever, respiratory distress, and hepatosplenomegaly, etc. Abnormal chest radiographs were found in 133 infants, of whom 83 cases showed miliary tuberculosis and multiple pulmonary nodules. Sixty eight infants died from among the 169 cases. The mortality dropped to 21.7% after treatment with anti-tuberculosis medication. The blood leukocyte count (P < 0.001), anti-tuberculosis treatment (P < 0.001), age of onset (P = 0.004), and presence of intracranial lesions (P < 0.001) affected the prognosis of congenital tuberculosis. CONCLUSIONS: The majority of infants with congenital tuberculosis onset within 2-3 weeks after delivery had no specific manifestations. Anti tuberculosis medication could reduce the mortality. The age of onset, presence of intracranial lesions, anti-tuberculosis treatment, specific image performances and leukocyte count were related to the prognosis of congenital tuberculosis. PMID- 21626716 TI - Discriminative properties of two predictive indices for asthma diagnosis in a sample of preschoolers with recurrent wheezing. AB - INTRODUCTION: It is important to predict if preschool children with recurrent wheezing will suffer from asthma in future years. To aid in this early identification, a number of asthma predictive scores have been reported, such as the asthma predictive index (API) and the PIAMA risk score. However, to the best of our knowledge, their predictive properties have not been evaluated in any lower- to middle-income country. MATERIALS AND METHODS: A prospective cohort study was carried out including preschoolers aged 1-3 years with recurrent wheezing who came to our Pediatric Pulmonary Unit in Bogota, Colombia. We collected the information required to complete the API index and the PIAMA risk score. At 5-6 years of age, the patients were contacted in order to determine if they were suffering from active asthma. We calculated the sensitivity, specificity, predictive values, and likelihood ratios (LR) of the API and PIAMA risk scores for the presence of active asthma at 5-6 years old. RESULTS: The mean age at recruitment of the 130 included patients was 27.2 +/- 5.9 months. The loose API yielded a sensitivity of 71.4% (95% CI: 50.0-86.2), specificity of 33.3% (95% CI 23.5-44.8), and positive predictive value of 23.8% (95% CI: 15.0 35.6). The stringent API yielded a sensitivity of 42.9% (95% CI: 24.5-63.5), specificity of 79.2% (95% CI 68.4-86.9), positive predictive value of 37.5% (95% CI: 21.2-57.3), and positive LR of 2.06. The PIAMA risk score yielded a sensitivity of 54.5% (95% CI: 42.6-66.0), specificity of 78.9 (95% CI: 66.7 87.5), positive predictive value of 75.0 (95% CI: 61.2-85.1), and positive LR of 2.59. CONCLUSIONS: Our results suggest that both indices can be used to predict asthma in preschoolers with recurrent wheezing in the context of a referral hospital. PMID- 21626717 TI - Cystic fibrosis related diabetes (CFRD)--the end stage of progressive insulin deficiency. AB - In cystic fibrosis, gradual pancreatic destruction causes progressive insulin deficiency, culminating in cystic fibrosis related diabetes (CFRD). As a consequence of insulin deficiency, elevated glucose levels can be detected (well before the diagnosis of CFRD), by continuous ambulatory subcutaneous interstitial fluid glucose monitoring or 30-min sampled oral glucose tolerance test (OGTT). Current diagnostic criteria for CFRD (based on 0 and 120-min OGTT blood glucose levels) were originally designed to forecast microvascular disease in type 2 diabetes, rather than CF-specific outcomes such as declining weight or lung function. In CF, decline in either weight or lung function predicts early mortality. Both may precede the diagnosis of CFRD by several years. Insulin, a potent anabolic hormone, is recommended treatment for CFRD, but use in earlier stages of insulin deficiency is not established. Conventional dosing (with four or more insulin injections per day) is burdensome and carries substantial risk of hypoglycemia. However, recent uncontrolled trials suggest that once-daily injection of intermediate or long-acting insulin improves weight and lung function, with minimal hypoglycemia risk, in CFRD and also in early insulin deficiency. It is plausible that insulin may be of greater benefit to respiratory function when given prior to the diagnosis of CFRD, after which structural lung disease may be irreversible. It is also plausible that early insulin treatment may prolong the lifespan of the remaining insulin-secreting beta-cells. Randomized controlled trials are now needed to determine whether or not current clinical practice should be altered toward the earlier commencement of insulin in CF. PMID- 21626718 TI - Cytokine response in pediatric patients with pandemic influenza H1N1 2009 virus infection and pneumonia: comparison with pediatric pneumonia without H1N1 2009 infection. AB - OBJECTIVES: We investigated serum cytokine levels in pediatric patients with pandemic influenza H1N1 2009 virus (H1N1) infection-pneumonia and in pediatric patients with pneumonia but without H1N1 infection, and examined correlations between cytokine levels and clinical/laboratory findings. METHODS: Fifty-seven cases of infection by H1N1 were confirmed by RT-PCR and enrolled. Of these 57 cases, 26 had a severe H1N1 infection (group 1), and 31 had a mild H1N1 infection (group 2). Sera from 18 cases with pneumonia without H1N1 infection (group 3) were used as controls. The serum levels of 10 cytokines were determined by multiplex assay. RESULTS: The serum levels of IFN-alpha, IL-6, and IP-10 were significantly higher in H1N1 infected cases than in group 3, and levels of IL-6 and IP-10 were significantly higher in group 1 than in group 2. The level of IL 10 was significantly higher in groups 1 and 3 than in group 2. However, levels of IFN-gamma and IL-17 were not significantly different between the three groups. IL 1beta, IL-4, and MIP-1alpha were not detectable in most patients. IP-10 and IL-6 levels were found to show negative correlations with lymphocyte count and oxygen saturation. CONCLUSIONS: We found higher levels of cytokines (IFN-alpha, IL-6, IP 10) of innate immunity than those of acquired immunity in pediatric H1N1 infection. Of the cytokines found to be increased in cases with H1N1 infection, IP-10 and IL-6 were found to be correlated with disease severity (lymphopenia and hypoxia). IP-10 and IL-6 may be important markers in pediatric H1N1 infection. PMID- 21626719 TI - The effect of astrocytes on the induction of barrier properties in aortic endothelial cells. AB - Construction of in vitro models of the blood-brain barrier (BBB) using primary brain microvascular endothelial cells (BMEC) is time intensive and not high throughput, in part due to a lack of culture purity, low yields, and cellular dedifferentiation after the first passage. This problem has created interest in the substitution of BMEC with immortalized brain endothelial cells (EC), or peripheral EC such as bovine aortic EC (BAEC). Many BBB models have focused on further inducing the brain and peripheral ECs by incorporating astrocyte back-to back or nonback-to-back cocultures. However, previous studies demonstrating induction effects of astrocytes on BAEC in back-to-back cocultures failed to recognize the extensive barrier properties of astrocytes alone, which can have a significant effect on interpreting the results. This manuscript reports the establishment of back-to-back and nonback-to-back cocultures between astrocytes and BAEC or BMEC (as a control) with primary focus on the properties of astrocytes alone and with a linear contrast statistical methodology to interpret the results. Transendothelial electrical resistance and permeability studies revealed that astrocytes can significantly increase the barrier tightening of BMEC by 167%, while having no effect on BAEC. Immunocytochemical studies also revealed the reorganization of BMEC occludin junctions in the presence of astrocytes, while indicating the absence of this junctional protein in BAEC. In contrast to a previous report, here the linear contrast statistical analysis revealed that observed decreases in permeability of BAEC in back-to-back cocultures is due to the addition of astrocytes' properties in series and not due to induction. PMID- 21626720 TI - Application of electrochemically produced aluminum hydroxide gel for prepurification of recombinant synthetic green fluorescent protein produced in tobacco leaves. AB - The use of recombinant proteins has increased greatly in recent years, as also have increased the number of techniques and materials used for their production and purification. Among the different types of bioreactors being studied, there is a general consensus among scientists that production in green plant tissues such as leaves is more feasible. However, the presence of chlorophyll and phenolic compounds in plant extracts, which can precipitate and denature the proteins besides damaging separation membranes and gels, makes this technology impracticable on a commercial scale. In the present work, the adsorption to electrochemically produced aluminum hydroxide gel was applied as a prepurification step for recombinant synthetic green fluorescent protein (sGFP), also referred to as enhanced green fluorescent protein, produced in Nicotiana benthamiana leaves. Removal efficiencies of 99.7% of chlorophyll, 88.5% of phenolic compounds, and 38.5% of native proteins from the N. benthamiana extracts were achieved without removing sGFP from the extracts. As electrochemical preparation of aluminum hydroxide gel is a cost-effective technique, its use can substantially contribute to the development of future production platforms for recombinant proteins produced in green plant tissues of pharmaceutical and industrial interest. PMID- 21626721 TI - Drug loading into and drug release from pH- and temperature-responsive cylindrical hydrogels. AB - Hydrogels that undergo deformation upon appropriate changes in pH or temperature have considerable promise as drug delivery vehicles. Drug uptake in swelling and nonswelling cylindrical hydrogels and drug release from these into a target fluid are investigated here. A mathematical model for hydrogel-solution composite, a composite of a distributed parameter system (cylindrical hydrogel) and a lumped parameter system (surrounding solution), is developed. The polymer network displacement in a swelling/deswelling hydrogel is described by a stress diffusion coupling model. The analytical solution for network displacement is used to predict solvent intake by swelling hydrogels, solvent efflux from deswelling hydrogels, and changes in pressure, porosity, and effective drug diffusivity. These in turn influence drug uptake during and after hydrogel swelling and drug release from hydrogel during and after deswelling. Numerical results illustrate benefits of hydrogel swelling for drug loading and merits of different modes of drug release. Drug uptake and drug release by temperature-responsive hydrogels are compared with those by hydrogels not subject to deformation. PMID- 21626722 TI - Performance of high intensity fed-batch mammalian cell cultures in disposable bioreactor systems. AB - The adoption of disposable bioreactor technology as an alternate to traditional nondisposable technology is gaining momentum in the biotechnology industry. Evaluation of current disposable bioreactors systems to sustain high intensity fed-batch mammalian cell culture processes needs to be explored. In this study, an assessment was performed comparing single-use bioreactors (SUBs) systems of 50 , 250-, and 1,000-L operating scales with traditional stainless steel (SS) and glass vessels using four distinct mammalian cell culture processes. This comparison focuses on expansion and production stage performance. The SUB performance was evaluated based on three main areas: operability, process scalability, and process performance. The process performance and operability aspects were assessed over time and product quality performance was compared at the day of harvest. Expansion stage results showed disposable bioreactors mirror traditional bioreactors in terms of cellular growth and metabolism. Set-up and disposal times were dramatically reduced using the SUB systems when compared with traditional systems. Production stage runs for both Chinese hamster ovary and NS0 cell lines in the SUB system were able to model SS bioreactors runs at 100-, 200 , 2,000-, and 15,000-L scales. A single 1,000-L SUB run applying a high intensity fed-batch process was able to generate 7.5 kg of antibody with comparable product quality. PMID- 21626723 TI - Dual salt precipitation for the recovery of a recombinant protein from Escherichia coli. AB - When considering worldwide demand for biopharmaceuticals, it becomes necessary to consider alternative process strategies to improve the economics of manufacturing such molecules. To address this issue, the current study investigates precipitation to selectively isolate the product or remove contaminants and thus assist the initial purification of a intracellular protein. The hypothesis tested was that the combination of two or more precipitating agents will alter the solubility profile of the product through synergistic or antagonistic effects. This principle was investigated through several combinations of ammonium sulfate and sodium citrate at different ratios. A synergistic effect mediated by a known electrostatic interaction of citrate ions with Fab' in addition to the typical salting-out effects was observed. On the basis of the results of the solubility studies, a two step primary recovery route was investigated. In the first step termed conditioning, post-homogenization and before clarification, addition of 0.8 M ammonium sulfate extracted 30% additional product. Clarification performance measured using a scale-down disc stack centrifugation mimic determined a four-fold reduction in centrifuge size requirements. Dual salt precipitation in the second step resulted in >98% recovery of Fab' while removing 36% of the contaminant proteins simultaneously. PMID- 21626724 TI - Estimating cell specific oxygen uptake and carbon dioxide production rates for mammalian cells in perfusion culture. AB - We present robust methods for online estimation of cell specific oxygen uptake and carbon dioxide production rates (q(O2) and q(CO2), respectively) during perfusion cultivation of mammalian cells. Perfusion system gas and liquid phase mass balance expressions for oxygen and carbon dioxide were used to estimate q(O2), q(CO2) and the respiratory quotient (RQ) for Chinese hamster ovary (CHO) cells in perfusion culture over 12 steady states with varying dissolved oxygen (DO), pH, and temperature set points. Under standard conditions (DO = 50%, pH = 6.8, T = 36.5 degrees C), q(O2) and q(CO2) ranges were 5.14-5.77 and 5.31-6.36 pmol/cell day, respectively, resulting in RQ values of 0.98-1.14. Changes to DO had a slight reducing effect on respiration rates with q(O2) and q(CO2) values of 4.64 and 5.47, respectively, at DO = 20% and 4.57 and 5.12 at DO = 100%. Respiration rates were lower at low pH with q(O2) and q(CO2) values of 4.07 and 4.15 pmol/cell day at pH = 6.6 and 4.98 and 5.36 pmol/cell day at pH = 7. Temperature also impacted respiration rates with respective q(O2) and q(CO2) values of 3.97 and 4.02 pmol/cell day at 30.5 degrees C and 5.53 and 6.25 pmol/cell day at 37.5 degrees C. Despite these changes in q(O2) and q(CO2) values, the RQ values in this study ranged from 0.98 to 1.23 suggesting that RQ was close to unity. Real-time q(O2) and q(CO2) estimates obtained using the approach presented in this study provide additional quantitative information on cell physiology both during bioprocess development and commercial biotherapeutic manufacturing. PMID- 21626725 TI - Proceedings of the Sixth Inter-Institute Workshop on Optical Imaging from Bench to Bedside. October 2009. Bethesda, Maryland, USA. PMID- 21626726 TI - Special issue to honor Erich Buchner on the occasion of his 65th birthday. PMID- 21626727 TI - Recent progress in therapeutic apheresis. Proceedings of the 1st Dresden International Symposium on Therapeutic Apheresis. September 4-5, 2009. Dresden, Germany. PMID- 21626728 TI - Proceedings of the VII Colloquium Chemiometricum Mediterraneum (CCM VII). June 21 14, 2010. Granada, Spain. PMID- 21626729 TI - Proceedings of the 2nd International Symposium on Chylomicrons in Disease-2 (ISCD2). June 2-5, 2010. Rotterdam, The Netherlands. PMID- 21626730 TI - Festschrift to honor Horst R. Thieme on the occasion of his 60th birthday. PMID- 21626731 TI - Festschrift in memory of Michael Stephen Feld: physics and biomedicine pioneer (1940-2010). PMID- 21626732 TI - Proceedings of the 12th Biennial Meeting on Laser Applications to Chemical, Security, and Environmental Analysis (LACSEA). January 31-February 3, 2010. San Diego, California, USA. PMID- 21626733 TI - Nip gossip in the bud. PMID- 21626734 TI - No place for placebos. PMID- 21626735 TI - Diving BAC into the pool with microarrays. PMID- 21626736 TI - Illuminating dark matter. PMID- 21626737 TI - Carbohydrates in sustainable development, Part 2 - A mine for functional molecules and materials. Preface. PMID- 21626738 TI - Knoevenagel reaction of unprotected sugars. AB - The Knoevenagel reaction of unprotected sugars was investigated in the 1950s using zinc chloride as promoter. The so-called Garcia Gonzalez reaction had been almost forgotten for 50 years, until the emergence of new water tolerant catalysts having Lewis acid behavior. The reaction was thus reinvestigated and optimal conditions have been found to prepare trihydroxylated furan derivatives from pentose or beta-tetrahydrofuranylfuran from hexoses with non-cyclic beta keto ester or beta-diketones. Other valuable compounds such as beta-linked tetrahydrobenzofuranyl glycosides or hydroxyalkyl-3,3,6,6,-tetramethyl 3,4,5,6,7,9-hexahydro-1H-xanthene-1,8(2H)-dione can be obtained using cyclic beta dicarbonylic derivatives. Apart from one report in the 1950s, the Knoevenagel reaction of unprotected carbohydrate in basic condition has been studied only in the mid-1980s to prepare C-glycosyl barbiturates from barbituric acids and, later on, from non-cyclic beta-diketones, beta-C-glycosidic ketones. The efficient method exploited to prepare such compounds has found an industrial development in cosmetics. PMID- 21626739 TI - Carbohydrate-based lactones: synthesis and applications. AB - The synthesis and uses of different kinds of carbohydrate-based lactones are described. This group of compounds includes aldonolactones, other related monocyclic lactones and bicyclic systems. The latter can arise from uronic acids, carboxymethyl ethers or glycosides, or from C-branched sugars. PMID- 21626740 TI - Heterogeneously-catalyzed conversion of carbohydrates. AB - Polyfunctionality of carbohydrates and their low solubility in conventional organic solvents make rather complex their conversion to higher value added chemicals. Therefore, innovative processes are now strongly needed in order to increase the selectivity of these reactions. Here, we report an overview of the different heterogeneously-catalyzed processes described in the literature. In particular, hydrolysis, dehydration, oxidation, esterification, and etherification of carbohydrates are presented. We shall discuss the main structural parameters that need to be controlled and that permit the conversion of carbohydrates to bioproducts with good selectivity. The conversion of monosaccharides and disaccharides over solid catalysts, as well as recent advances in the heterogeneously-catalyzed conversion of cellulose, will be presented. PMID- 21626741 TI - Palladium-catalyzed telomerization of butadiene with polyols: from mono to polysaccharides. AB - The telomerization of butadiene with alcohols is an elegant way to synthesize ethers with minimal environmental impact since this reaction is 100% atom efficient. Besides telomerization of butadiene with methanol and water that is industrially developed, the modification of polyols is still under development. Recently, a series of new substrates has been involved in this reaction, including diols, pure or crude glycerol, protected or unprotected monosaccharides, as well as polysaccharides. This opens up the formation of new products having specific physicochemical properties. We will describe recent advances in this field, focusing on the reaction of renewable products and more specifically on saccharides. The efficient catalytic systems as well as the optimized reaction conditions will be described and some physicochemical properties of the products will be reported. PMID- 21626742 TI - Enzymatic processing of bioactive glycosides from natural sources. AB - A number of biologically active natural products are glycosides. Often, the glycosidic residue is crucial for their activity. In other cases, glycosylation only improves their pharmacokinetic parameters. Enzymatic modification of these glycosides - both extension of the glycoside moiety and its selective trimming - is advantageous due to their selectivity and mildness of the reaction conditions in the presence of reactive and sensitive complex aglycones. Enzymatic reactions enable the resulting products to be used as "natural products", e.g., in nutraceuticals. This chapter concentrates on naturally occurring glycosides used in medicine but also in the food and flavor industry (e.g., sweeteners). Both "classical" and modern methods will be discussed. PMID- 21626743 TI - Synthetic polymers from readily available monosaccharides. AB - The low degradability of petroleum-based polymers and the massive use of these materials constitute a serious problem because of the environmental pollution that they can cause. Thus, sustained efforts have been extensively devoted to produce new polymers based on natural renewing resources and with higher degradability. Of the different natural sources, carbohydrates stand out as highly convenient raw materials because they are inexpensive, readily available, and provide great stereochemical diversity. New polymers, analogous to the more accredited technical polymers, but based on chiral monomers, have been synthesized from natural and available sugars. This chapter describes the potential of sugar-based monomers as precursors to a wide variety of macromolecular materials. PMID- 21626744 TI - Synthesis and applications of ionic liquids derived from natural sugars. AB - Aiming to develop environmentally compatible chemical syntheses, the replacement of traditional organic solvents with ionic liquids (ILs) has attracted considerable attention. ILs are special molten salts with melting points below 100 degrees C that are typically constituted of organic cations (imidazolium, pyridinium, sulfonium, phosphonium, etc.) and inorganic anions. Due to their ionic nature, they are endowed with high chemical and thermal stability, good solvent properties, and non-measurable vapor pressure. Although the recovery of unaltered ILs and recycling partly compensate their rather high cost, it is important to develop new synthetic approaches to less expensive and environmentally sustainable ILs based on renewable raw materials. In fact, most of these alternative solvents are still prepared starting from fossil feedstocks. Until now, only a limited number of ILs have been prepared from renewable sources. Surprisingly, the most available and inexpensive raw material, i.e., carbohydrates, has been hardly exploited in the synthesis of ILs. In 2003 imidazolium-based ILs were prepared from o-fructose and used as solvents in Mizoroki-Heck and Diels-Alder reactions. Later on, the first chiral ILs derived from sugars were prepared from methyl D-glucopyranoside. In the same year, a family of new chiral ILs, obtained from commercial isosorbide (dianhydro-D glucitol), was described. A closely related approach was followed by other researchers to synthesize mono- and bis-ammonium ILs from isomannide (dianhydro-D mannitol). Finally, a few ILs bearing a pentofuranose unit as the chiral moiety were prepared using sugar phosphates as glycosyl donors and 1-methylimidazole as the acceptor. PMID- 21626745 TI - Carbohydrates in sustainable development, Part 1. Renewable resources for chemistry and biotechnology. Preface. PMID- 21626746 TI - Sucrose: A prospering and sustainable organic raw material. AB - Sucrose (alpha-D-glucopyranosyl-(1-->2)-beta-D-fructofuranoside) is an inexpensive chemical produced by sugar cane and sugar beet cultivation. Chemical and/or biochemical transformations convert it into highly valuable synthetic intermediates such as 5-hydroxymethylfurfural (HMF), bioethylene, 1,2-propylene glycol and levulinic acid. Sucrose can also be converted into biodegradable polymers such as polyesters and polyurethanes, as well as into novel carbohydrates such as isomaltulose, trehalulose, inulin, levan, Neo-amylose, and dextran, highly valuable additives for food and cosmetics and materials for separation and purification technologies. PMID- 21626747 TI - Sucrose-utilizing transglucosidases for biocatalysis. AB - Sucrose-utilizing transglucosidases are valued tools in chemistry to generate glycodiversification. Not only do these enzymes use as substrate an abundant agroresource, sucrose, but they also share a remarkable versatility regarding the acceptor substrate, allowing the structurally-controlled synthesis of diverse glucosylated products. Latest research has demonstrated the potential of enzyme engineering to tailor novel sucrose-utilizing transglucosidases that give access to original carbohydrate-based structures. This chapter gives an overview of the recent achievements in biocatalysis using these enzymes. PMID- 21626748 TI - Difructose dianhydrides (DFAs) and DFA-enriched products as functional foods. AB - This review provides an overview of the current status of the chemistry and biology of di-D-fructose dianhydrides (DFAs) with a focus on their potential as functional foods. The history of this family of cyclic ketodisaccharides has expanded for almost 100 years and offers a paradigmatic example of artificial synthetic molecules that were identified as natural products later on and finally encountered in our own table. Issued from fundamental investigations on the reactivity of carbohydrates in strongly acidic media, DFAs remained laboratory curiosities for decades. Early reports on their isolation from plants raised doubts, until the formation of some DFA representatives by the action of microorganisms on fructans was reported in the middle 1980s. Since then, research on DFAs has run in parallel in the areas of microbiology and carbohydrate chemistry. Evidence of the potential of these compounds as functional food was accumulated from both sides, with the development of biotechnological processes for mass production of selected candidates and of chemical methodologies to prepare DFA-enriched products from sucrose or inulin. In 1994 a decisive discovery in the field took place in the laboratory of Jacques Defaye in Grenoble, France: the presence of DFAs in a commercial sucrose caramel was evidenced in a quite significant 18% mass proportion! The development of an efficient analytical protocol for DFAs and the stereoselective synthesis of individual standards allowed one to demonstrate that DFAs and their glycosylated derivatives (glycosyl-DFAs) are universally formed during caramelization reactions. They are not potential food products; they have actually always been in our daily food. Most important, they seem to exert beneficial effects: they are acariogenic, low-caloric, and promote the growth of beneficial microflora in the gut. Most recent evidence indicates that DFAs can even protect the intestinal tract against agressive agents favor the assimilation of antioxidants, and act as a drug-like food for the treatment of colon ailments such as inflammatory bowel disease (Crohn disease). The development of efficient methodologies for the preparation of DFA-enriched caramels, compatible with the food and agricultural industry regulations, may lead to new natural functional foods and nutraceuticals based on DFAs in the near future. PMID- 21626749 TI - Development of agriculture left-overs: fine organic chemicals from wheat hemicellulose-derived pentoses. AB - This review is dedicated to wheat hemicelluloses and its main components D-xylose and L-arabinose as raw materials for fine organic chemistry. The context of the wheat agro-industry, its by-products, and extraction and hydrolysis of hemicelluloses to produce the pentoses are considered. The straightforward preparation of pentose-based surfactants, their properties, and their situation in the field of carbohydrate-based surfactants are addressed. Multistep transformations of pentoses are also described, first from a methodology point of view, with the aim of producing multifunctional enantiopure building-blocks, then considering targeted natural and/or bioactive products. Selected reactions of furfural, an important dehydration product of pentoses, are also presented. PMID- 21626750 TI - Cellulose and derivatives from wood and fibers as renewable sources of raw materials. AB - Cellulose is the most important biopolymer in Nature and is used in preparation of new compounds. Molecular structure of cellulose is a repeating unit of beta-D glucopyranose molecules forming a linear chain that can have a crystallographic or an amorphous form. Cellulose is insoluble in water, but can dissolve in ionic liquids. Hemicelluloses are the second most abundant polysaccharides in Nature, in which xylan is one of the major constituents of this polymer. There are several sources of cellulose and hemicelluloses, but the most important source is wood. Typical chemical modifications are esterifications and etherifications of hydroxyl groups. TEMPO-mediated oxidation is a good method to promote oxidation of primary hydroxyl groups to aldehyde and carboxylic acids, selectively. Modified cellulose can be used in the pharmaceutical industry as a metal adsorbent. It is used in the preparation of cellulosic fibers and biocomposites such as nanofibrils and as biofuels. PMID- 21626751 TI - Olive pomace, a source for valuable arabinan-rich pectic polysaccharides. AB - Cell wall polysaccharides account for nearly one third of olive pomace dry matter produced by the environment friendly biphasic system. These polysaccharides are mainly cellulose, glucuronoxylans, and arabinan-rich pectic polysaccharides, in equivalent proportions. The structural features of pectic polysaccharides are unique concerning the arabinan moiety due to the occurrence of a beta-(1-->5) terminally-linked arabinose residue. This odd feature tends to disappear with olive ripening and can be used as a diagnostic tool in the evaluation of the stage of ripening of this fruit, as well as a marker for the presence of olive pulp in matrices containing pectic polysaccharides samples. These pectic polysaccharides have the ability to form elastic gels with calcium. The critical gelling calcium and galacturonic acid concentrations are higher than that observed for commercial citrus low-methoxyl pectic material. Nevertheless, they present a syneresis occurring for much higher calcium concentration and, consequently, show a much larger zone in which homogeneous gels are formed. In addition, olive pomace pectic polysaccharides gels are more resistant to temperature than the low-methoxyl pectin/calcium gels. These properties show that olive pomace can be a potential source of gelling pectic material with useful properties for particular applications. PMID- 21626752 TI - Oligomannuronates from seaweeds as renewable sources for the development of green surfactants. AB - The development of surfactants based on natural renewable resources is a concept that is gaining recognition in detergents, cosmetics, and green chemistry. This new class of biodegradable and biocompatible products is a response to the increasing consumer demand for products that are both "greener", milder, and more efficient. In order to achieve these objectives, it is necessary to use renewable low-cost biomass that is available in large quantities and to design molecular structures through green processes that show improved performance, favorable ecotoxicological properties and reduced environmental impact. Within this context, marine algae represent a rich source of complex polysaccharides and oligosaccharides with innovative structures and functional properties that may find applications as starting materials for the development of green surfactants or cosmetic actives. Thus, we have developed original surfactants based on mannuronate moieties derived from alginates (cell-wall polyuronic acids from brown seaweeds) and fatty hydrocarbon chains derived from vegetable resources. Controlled chemical and/or enzymatic depolymerizations of the algal polysaccharides give saturated and/or unsaturated functional oligomannuronates. Clean chemical processes allow the efficient transformation of the oligomers into neutral or anionic amphiphilic molecules. These materials represent a new class of surface-active agents with promising foaming/emulsifying properties. PMID- 21626753 TI - From natural polysaccharides to materials for catalysis, adsorption, and remediation. AB - Polysaccharides display most of the properties needed for applications in catalysis, adsorption or remediation. Requisites common to these applications are appropriate surface functions to ensure substrate-material interactions, accessibility of the functional groups, and proper shaping for easy manipulation. Natural polysaccharides are well known as supports for enzymatic catalysts and gelling agents in aqueous phase, due to the high level of dispersion of hydrocolloids. However, they suffer from diffusional limitations in the dry state, due to the low surface area of the dried materials generally used, xerogels or lyophilized solids. This chapter deals with the proper methods to prepare dry materials which retain the dispersion of the polymer hydrogel, namely polysaccharide aerogels. The materials whose properties are described here are stable in most organic solvents and present numerous and diverse surface functionalities (like hydroxy, carboxy, or amino groups). Shaping and appropriate drying of gelling polysaccharides provide a new opportunity to obtain materials from one of the less energy-intensive sources of biomass. Their application in catalysis and adsorption could open substantial markets for products of seaweed harvesting and coproducts of the seafood industry. PMID- 21626754 TI - Systems biology. PMID- 21626755 TI - Update 1 of: Mass accommodation and chemical reactions at gas-liquid interfaces. PMID- 21626756 TI - Proceedings of the 30th Annual Conference of the International Society for Clinical Biostatistics. August 23-27, 2009. Prague, Czech Republic. PMID- 21626759 TI - [Factors influencing early results of femoro-femoral crossover bypass]. AB - INTRODUCTION: Femoro-femoral crossover bypass is an extraanatomic reconstruction used for revascularization of lower limb with contralatateral femoral artery as an inflow vessel, and the graft placed in the suprapubic region. We perform this procedure when anatomic reconstruction is not possible or is contraindicated. OBJECTIVE: To analyze the influence of different risk factors on early patency of femoro-femoral crossover bypass. METHODS: This retrospective study analyzed the results of 88 femoro-femoral bypass grafting during an 11-year period. There were 66 (75%) males and 22 (25%) females of average age 64.93 years (42-79 years). In 76 patients the operations were performed due to critical limb ischemia. Revascularization was urgent in 12 patients, while 76 patients were elective. Dacron prosthesis was used in 81 patients, while PTFE was used in 7 patients. Statistical analysis was made by logistic regression. RESULTS: During hospitalisation the graft remained patent in 82 patients, and graft thrombosis occurred in 6 patients. Limb salvage rate was 90.91%. Early morbidity rate (within the first postoperative month) was 13.64%, while early mortality rate was 4.55%. Using logistic regression we established that early graft patency was statistically more significant in males (p < 0.05). Age (p = 0.07) and hypertension (p = 0.08) appeared to be predicting influence of the graft patency on the border of the accepted statistical significance level. CONCLUSION: Femoro femoral crossover bypass is a good alternative for revascularization in high risk patients for standard anatomic reconstructions due to comorbid conditions or local problems. PMID- 21626760 TI - [Breech presentation at term: caesarean section or vaginal delivery?]. AB - INTRODUCTION: Term breech presentation occurs in 3-4% of all deliveries. Most obstetricians are completely sure how to end delivery when the foetus is in breech presentation, by caesarean section as the only option. OBJECTIVE: The main goal of the paper was to present the method of delivery for term breech presentation analyzed at the Hospital of Gynaecology and Obstetrics and Clinical Centre in Kragujevac, and parameters that influenced the decision on the method of choice to perform delivery with the foetus in breech position. METHODS: During a three-year prospective study, the course and outcome of all term breech deliveries were under followup. The study involved only deliveries in breech presentation, with a trial of labour ending by vaginal delivery, while elective caesarean section due to breech presentation of the foetus was not included in the study. The following parameters were compared: body mass, newborn's first minute Apgar score and head circumference, gestational maturity, gestational age, delivery duration, maternal level of education and maternal parity. RESULTS: Of total 6,470 deliveries, 653 (10.10%) were finalized by caesarean section. Of these, there were 202 (3.12%) term breech presentations, of which 72 (35.64%) women had caesarean and 130 (64.36%) vaginal delivery. A difference was detected in newborn's body mass and head circumference, delivery duration, maternal level of education and parity between pregnancies terminated surgically in comparison to vaginal delivery, while pregnancy duration, maternal age, first minute Apgar score, and most significantly, perinatal morbidity did not show any difference regardless of the method of delivery for term breech presentation. CONCLUSION: With respect of all known parameters, vaginal delivery in breech presentation is also accepted. PMID- 21626761 TI - [Ultrasonography of the first metatarsophalangeal joint in gout]. AB - INTRODUCTION: About one half of the first gout attacks occur in the first metatarsophalangeal joint (MTPJ1); in the disease course this joint is practically inevitably affected. Radiographic evidence of bone erosions is the indication for hypouricaemic therapy in order to prevent joint destruction and nephropathy. Advantages of ultrasonography (US) comparing to conventional x-ray findings in depicting early bone erosions in various inflammatory arthropathies have been demonstrated by several studies. OBJECTIVE: The aims of this study were to compare US and x-ray findings in the detection of MTPJ1 erosions in patients with gout, to correlate sonographic and clinical features, and to detect possible characteristic sonographic features of gout. METHODS: Thirty patients (60 MTPJ1) with primary gout (ACR) and 10 age-matched control subjects (20 MTPJ1) with different inflammatory arthropathies were clinically evaluated. Standard dorsiplantar weight bearing and lateral weight bearing x-ray views of both feet were taken. US was performed and interpreted by an independent sonographer on the presence of bone erosions, synovial fluid, synovial hypertrophy, Doppler signal and hyperechoic spots. Statistical analysis was performed (Spearman and Pearson correlation coefficient, Wilcoxon and chi2 test.) RESULTS: Twenty-four studied MTPJ1 had evidence of erosions, 17 only on US and seven both on x-ray and on US (Z = -4.123; p = 0.000). US findings showed that hyperechoic spots were the most prominent feature of gouty MTPJ1 (chi2 = 40.909; p = 0.000), followed by erosions and synovial fluid presentation. CONCLUSION: US of MTPJ1 in gout discovers significantly more erosions than x-ray, which may have therapeutic implications. The evidence of hyperechoic spots (surrogate crystals) of the different size, number and orientation is a major sonographic feature of the MTPJ1 in gout, which may be of importance in the diagnosis of certain cases (low serum urate, unavailable synovial fluid or the urate crystals absence). PMID- 21626762 TI - [Quality of life in patients with chronic hepatitis C]. AB - INTRODUCTION: Hepatitis C virus often causes chronic liver disease reducing physical, mental and social functions in these patients. OBJECTIVE: The aim of this investigation was analysis of the quality of life in chronic hepatitis C patients compared to patients with other chronic liver diseases and healthy population, as well as investigation of the influence of socio-demographic factors on the quality of life in patients with chronic hepatitis C. METHODS: A generic Short Form-36 (SF-36) questionnaire and Chronic Liver Diseases Questionnaire (CLDQ) were used in this prospective study for the investigation of the quality of life in 160 patients (100 patients with chronic hepatitis C, 30 patients with chronic hepatitis B, 30 patients with non-viral chronic liver diseases) and 50 healthy controls. RESULTS: Reduced quality of life was noted in patients with chronic hepatitis C patients in comparison with healthy controls (p = 0.00). Significant differences in SF-36 were found between patients with chronic hepatitis C and B regarding physical functions, activity, physical pain and emotional functions. Multivariate linear regression analysis revealed ages below 50 years as the most important positive variable in chronic hepatitis C patients for total score of the quality of life and physical component score (B = 14.5; SE = 5.16; p = 0.049; and B = 16.4; SE = 5.94; p = 0.003, respectively). The most important positive variable for the mental component of the quality of life was male gender (B = 15.3; SE = 5.81; p = 0.003). CONCLUSION: Quality of life is reduced in patients with chronic hepatitis C in comparison with healthy population. The quality of life in chronic hepatitis C patients is better than in patients with other non-viral chronic liver diseases. In comparison with patients with chronic hepatitis B, reduction in some domains of the quality of life is noted. Younger age is the most predictable group for the lowest damage of the total quality of life in patients with chronic hepatitis C patients among other sociodemographic characteristics of these patients. PMID- 21626763 TI - [The influence of sepsis as a complication after trauma on immune response to injury]. AB - INTRODUCTION: Mortality rate in trauma complicated with sepsis is exceeding 50%. Outcome is not determined only by infection or trauma, but also by the intensity of immuno-inflammatory response. OBJECTIVE: The aim of this study was to determine the influence of sepsis on the immuno-inflammatory response, in the group of 35 traumatized men, of which in 25 cases trauma was complicated with sepsis. METHODS: Cytokines were measured by ELISA test in plasma. Blood samples were drown on the first, third and fifth day after ICU admission. RESULTS: Proinflammatory cytokine IL-8 was 230-fold higher in trauma + sepsis group (1148.48 vs. 5.05 pg/ml; p < 0.01), and antiinflammatory cytokine IL-ra was 4 fold higher (1138.3 vs. 310.05 pg/ml; p < 0.01), whereas IL-12 and IL-4 showed no significant difference between the groups. CONCLUSION: We concluded that sepsis, as a complication after trauma, drastically enhances immuno-inflammatory response to insult, as indicated by IL-8 and IL-ra, but not IL-12 and IL-4. PMID- 21626764 TI - [Biometeorological influence on motorcyclists and bicyclists trauma]. AB - INTRODUCTION: Biometeorological circumstances have great influence on all traffic participants, especially on reflexes, movement coordination and perception ability, increasing the number of accidents. Motocyclists and bicyclists are specific participants in traffic, so their traumatism has its own special characteristics. OBJECTIVE: The aim of this study was to establish biometeorological influence on motorcyclists and bicyclists accidents. METHODS: Comparative analysis of everyday biometeorological phases for the city of Kragujevac determinated by the Republic Hydrometeorological Service of Serbia and motorcyclists and bicyclists accident evidence received from Kragujevac traffic police for the period 2004-2008. RESULTS: There were 5,180 traffic accidents in this period, with 180 accidents in which 77 motorcyclists and 116 bicyclists were knocked down. Most of the accidents were in 2008 (53) and the least in 2005 (28). Most accidents occurred during August (28) and between 19.00 and 20.00 hours (18). There were no accidents during January, between 04.00 and 06.00 hours. There were 85% accidents in the urban area. Most of 169 males were aged 16-20 and 21-25 years (20 and 21). Most of 24 females (6) were aged 11-15 years. Those who caused the accident mostly sustained injuries due to improper speed limit (31), while the victims due to the disrespect of street-crossing priorities (32). Forty one motorcyclists and 85 bicyclists suffered mild body injuries. Thirty-one motorcyclists and 27 bicyclist suffered severe injuries. Five motorcyclists and four bicyclists ended lethally. Most accidents happened in biometeorological phases 9 (63), 4 (32) and 1 (31). CONCLUSION: Statistical analysis showed a significant correlation between accidents and biometeorological phases, with most accidents occurring during stabile warm weather and rapid penetrations of cold fronts, and with a sudden switch of weather conditions. PMID- 21626765 TI - [Risk factors for syphilis infection in the South Backa blood donor population]. AB - INTRODUCTION: Syphilis is one of infectious diseases that can be transmitted by blood transfusion. Determining the frequency of syphilis among blood donors and the relevant risk factors is an important aspect of safe blood transfusion. OBJECTIVE: To determine the frequency and the trend of syphilis infection among blood donors in South Backa Region, to classify the confirmed infection and to describe demographic characteristics of seropositive blood donors and their risk factors. METHODS: This retrospective study analyzed results of routine EIA syphilis screening and blood donor data from South Backa Region between 2004 and 2008. RESULTS: Thirteen out of 153,431 tested blood units were confirmed as syphilis-infected.The study found that the overall frequency of infection was 8.47 per 100,000 blood donations, with a trend increase of 2.4%; there was a statistically significant (p < 0.001) higher frequency among first-time blood donors (51.93/100,000) than regular ones (2.84/100 000). The highest prevalence was noted in donors aged over 45 years. All cases were classified as past syphilis infections. Post-donation interviews revealed exposure risk in 69.23% of the donors. CONCLUSION: A higher frequency of past infections detected in older donors is probably due to a higher prevalence and incidence of syphilis in the past. Further continual epidemiologic follow-up of syphilis in blood donors is crucial in obtaining safe blood. PMID- 21626766 TI - [Mineral waters of the Pannonian basin spas in the Republic of Serbia]. AB - INTRODUCTION: The fact itself that thermo-mineral waters and mud have healing effects has always attracted attention throughout the history to exploit, explore and study their benefit on the human body. Modern lifestyle and the speed of life endanger man's psycho-physical health. This is why people more often return to old time proven values, the nature and natural health resorts. OBJECTIVE: To establish hydro-geological conditions for the formation of mineral water and to summarize their balneological characteristics in spas, i.e., in rehabilitation centres of the Pannonian basin of the Republic of Serbia, where underground waters are still actively exploited for balneotherapy. METHODS: By retrospective descriptive analysis, a recapitulation of hydro-geological conditions for the formation of mineral waters was made and their balneological characteristics were established in eight spas of the Pannonian basin. RESULTS: The healing spas of the Pannonian basin are predominated by HCO3 (2.9 g/l - 4.6% milival), iodic (up to 6.5 mg/l), slightly alkaline (pH up to 8.1) thermal-mineral water (temperature up to 72 degrees C, bounty to 36.6 l/s) with a significant content of Br (up to 8.1 mg/l), Fe (to 6.0 mg/l), metaboron (up to 60 mg/l) and metasilicon acid (up to 95 mg/l). They are used for external application, bathing and showering. CONCLUSION: Once the Pannonian Sea (the Paratethys), today a wide plain terrain is a tectonic depression of the lower Pannonian pont age with compact type aquifers. In the geological column of sedimentary rocks a large underground aquifer was formed with free water of high mineralization, high temperature and geothermal properties above the average in relation to the European hydrogeological standards. Therefore, the Pannonian basin can be rightly called a thermal valley with the predominance of sodium hydro-carbonate (alkaline) iodine healing water of enviable abundance and reserves. PMID- 21626767 TI - [The role of psychic factors in the pathogenesis of bronchial asthma]. AB - Bronchial asthma is a chronic inflammatory disorder of the airways in which many cells play a role, in particular mast cells, eosinophils, neutrophils, T lymphocytes and epithelial cells. In susceptible individuals this inflammation causes recurrent episodes of wheezing, breathlessness, chest tightness and cough, particularly at night and/or in the early morning. These symptoms are usually associated with variable and extensive limitations of airflow in the bronchi reversible spontaneously or by treatment. It has been shown that restrain of the effectors of stress response participate in the pathogenesis of bronchial asthma. Anger that is not expressed and frustrations may activate the limbic stress pathway. Thus, the released neurotransmitters followed by excitation thus causing psychogenic (mental or emotional) stress. It is also known that emotional stress may be responsible for the exacerbation of asthma. Namely, pronounced emotions cause hyperventilation and hypocapnia inducing bronchospasm. Certain psychological personality features are related to adaptive or inadequate body response to numerous life events. Thus, until the beginning of the last century, bronchial asthma was referred to as asthma nervosa, because clinicians clearly observed the psychological profile of patients with predominant fear of asphyxia and recurrent attacks of paroxysmal dyspnoea. Besides, increased sensitivity, repression of aggressive feelings and expressive empathy have been identified as the most frequent psychological characteristics of asthmatic patients. However, scientists are still far from a full understanding of bronchial asthma pathogenesis. The contribution of psychic factors has become meaningful in the understanding of the development of bronchial asthma. Having in mind that in the majority of patients asthma is a lifelong condition, there is a hope that further investigations of bronchial asthma psychogenesis will improve prevention and treatment of this disease. PMID- 21626768 TI - [Operation of traumatic cataract with metal foreign body in the lens]. AB - INTRODUCTION: We present a patient with perforative eye injury, a metal foreign body in the lens and traumatic cataract. The paper emphasises the importance of phacoemulsification in case of patients with a traumatic cataract and the presence of a metal foreign body. CASE OUTLINE: A 41-year-old patient had a perforative wound of the cornea caused by a metal foreign body that also perforated the anterior lens capsule and remained in the paracentral anterior part of the lens. The injury, which happened upon hitting of a hammer against a metal object, showed the presence of a tangential wound of the cornea adapted edges, and a formed anterior eye chamber. The presence of a metal spear-shaped foreign body was partly inside the anterior eye chamber and partly in the central area of the lens. The visual acuity of the injured eye was 0.2. The technique of removing the foreign body out of the lens and the phacoemulsification of the lens with the implantation of intraocular lens is presented. The paracentral wound on the cornea was not sutured because of well-adapted edges. On the first postoperative day there were no inflammatory signs, and best uncorrected visual acuity of 0.8. CONCLUSION: Posttraumatic cataracts with a metal foreign body in the lens require operative treatment in order to remove the foreign body, phacoemulsification and implantation of artificial lens. Because of anterior capsule lesion, special care should be taken in regard to anterior capsulorhexis and appropriate hydrodisection. Ocular hypotonia and possible damage of the posterior capsule in some cases can make phacoemulsification more difficult to perform. PMID- 21626769 TI - [Adams-Oliver syndrome: case report]. AB - INTRODUCTION: Adams-Oliver syndrome is characterized by congenital aplasia of the vertex skin of the skull in combination with skull and transverse limb defects. CASE OUTLINE: We presented a 5-month old female infant with Adams-Oliver syndrome manifested as cutis marmorata, dilated scalp veins and limb reduction defects. Clinical manifestation also included thumb hypoplasia and extreme hypoplasia of other fingers, with agenesis of all toes on both feet. Echocardiogram revealed foramen ovale apertum. Venography showed dilated malformed scalp and neck veins, predominantly on the right side. On the basis of the clinical features and extended investigation we confirmed Adams-Oliver syndrome in the presented patient. CONCLUSION: We recommended prenatal diagnosis in case of future pregnancies, ultrasound examination, and follow-up of foetal anomalies. PMID- 21626770 TI - [Wound infection after carotid artery endarterectomy caused by methicillin resistant Staphylococcus aureus: case report]. AB - INTRODUCTION: Carotid endarterectomy is a safe and reliable surgical procedure to treat the narrowing of the internal carotid artery greater than 70%. One of the most serious complications of surgical wound after endarterectomy is an infection caused by methicillin-resistant Staphylococcus aureus (MRSA) which is accompanied with high morbidity and mortality. CASE OUTLINE: A 62-year-old woman developed a graft infection caused by MRSA after a carotid endarterectomy with synthetic graft placement. Two years after unsuccessful antibiotic and local wound treatment, we replaced the infected graft by an autologous saphenous vein conduit. CONCLUSION: The case suggests that only by aggressive surgical approach with replacement of infected graft a successful outcome can be achieved in such a rare but potentially fatal post-operative complication. PMID- 21626771 TI - [Progression of nodal marginal zone lymphoma into diffuse large B cell lymphoma in a patient with Sjogren's syndrome]. AB - INTRODUCTION: Sjogren's syndrome is a chronic autoimmune disorder carrying the risk of the development of non-Hodgkin's lymphoma, most frequently marginal zone lymphoma. CASE OUTLINE: A 66-year-old male patient with Sjogren's syndrome, after a year of the disease, developed a nodal marginal zone lymphoma with lymphoma cells in peripheral blood which had the following immunophenotype: CD19, CD20, CD22, CD19/kappa, CD79b+. After six cycles of chemotherapy according to CHOP protocol (cyclophosphamide, doxorubicin, vincristine and prednisone) disease remission was achieved lasting four months, followed by enlargement of lymph nodes in all areas (generalized lymphadenopathy), splenomegaly and enlargement of the right parotid gland. Bone marrow biopsy and histology confirmed lymphoma of the same morphologic and immunohistochemic profile. Biopsy of a very enlarged hard right parotid gland, by using histology and immunohistochemistry, showed lymphoid tumour tissue with blast appearance and a number of nucleoli corresponding to centroblasts and less to immunoblasts. Immunophenotypes of these cells were as follows: CD79alfa+, CD20+, CD3-, bcl-2-; proliferative activity measured with KI-67 was high rating 60%. Histology and immunohistochemistry showed the co-existence of a diffuse large B cell lymphoma with marginal zone lymphoma. In spite of aggressive chemotherapy treatment according to protocol ESHAP (Vepesid 200 mg i.v. on 1st and 2nd day and 100 mg on 3rd, 4th and 5th day; Cisplatin 20-20-10 mg on 1st to 4th day) the disease showed a progressive course. CONCLUSION: In patients with Sjogren's syndrome, the possibility of lymphoma should be kept in mind and in suspected cases timely diagnostic and therapeutic measures should be undertaken. PMID- 21626772 TI - [Multifoetal pregnancies complicated by reversed arterial perfusion: report of four cases]. AB - INTRODUCTION: Twin reversed arterial perfusion syndrome (TRAP) or acardiac anomaly presents a rare and severe complication affecting monochorionic multiple pregnancies occurring in 1 per 35,000 pregnancies or 1 per 100 of monozygotic twins. We report four cases of multiple pregnancies with TRAP diagnosed over the last 2 years, which were under check-up and treatment during the entire pregnancy course finalized by delivery. OUTLINE OF CASES: In two cases pregnancies were trigeminal, and other two geminal, with acardia of one foetus, by anceps type in three and acephalus in one. Expectant management was initially done in all cases. The relation of acardiac and donor abdominal circumference was below 50% and negative signs of the donor's cardiac failure were detected in two cases, so that expectative management was continued until delivery. Due to obstetric indications, a cesarean section was performed on the 33rd and 34th gestational week, and healthy newborns were delivered. Another two acardiac twins had abdominal circumference ratio between the acardiac fetus and the donor of over 50%, and signs of congestive heart failure in the donor, which indicated invasive therapeutic procedure with absolute alcohol chemisclerosis; it was done by alcohol injection into the acardiac abdominal aorta which interrupted the retrograde blood-flow into the acardiac umbilicus. In the first case a successful intervention was performed at 29th gestational week, but after 12 hours distress signs were registered. The delivery was finalized by a cesarean section, and a live donor was delivered. In the second case the intervention was successfully done at the 20th gestational week, and after 96 hours from the intervention lethal ending of the donor was registered. CONCLUSION: Bearing in mind that the occurrence of TRAP is rare, it is necessary to sum-up experiences from a larger number of centres to determine efficient therapeutic procedure. PMID- 21626773 TI - [Prevention of oral diseases in children with acute leukaemia]. AB - Oral manifestations are often present in all types of leukaemia, and are particularly severe in children. Oral diseases may result as complications of aggressive cytotoxic therapy. On the other hand, oral diseases that are already present in the oral cavity could be aggravating the patient's general condition and lead to serious life-threatening infections. The aim of this paper was to highlight the importance of preventive measures in maintaining oral health, as well the measures for controlling oral disorders onset during the course of illness and applied therapy in children with acute leukaemia. All preventive measures that include proper, regular hygiene and intervention depend on the patient's haematological status and general health condition, and are performed with the consultation and approval of the haematologists-in-charge. The preventive program can be implicated in three stages; before the beginning of cytotoxic therapy, during active cytotoxic therapy and after cytotoxic therapy. Having in mind that oral disease can influence general treatment in children with acute leukaemia dramatically, developing protocols on the implementation of all measures of oral prevention and treatment is mandatory. PMID- 21626774 TI - [Heart failure in haemodialysis patients: evaluation and treatment]. AB - Cardiovascular diseases are the leading cause of death in patients on haemodialysis. Cardiovascular mortality rate in these patients is approximately 9% per year, with the highest prevalence of left ventricular hypertrophy, ischemic heart disease and congestive heart failure being the most frequent cardiovascular complications. Risk factors for cardiac failure include hypertension, disturbed lipid metabolism, oxidative stress, microinflammation, hypoalbuminemia, anaemia, hyperhomocysteinemia, and increased concentration of asymmetric dimethylarginine, increased shunt blood flow and secondary hyperparathyroidism. Diagnostic strategy for early detection of patients with increased risk for the development of asymptomatic disturbances of systolic and diastolic left ventricular function should include echocardiografic examination, tests for determining coronary vascular disease, as well as tests of myocardial function (BNP, Nt-proBNP). Early detection of patients with a high risk of congestive heart failure enables timely implementation of adequate therapeutic strategy to provide high survival rate of HD patients. PMID- 21626775 TI - [Nation's health, public health services, or public health]. AB - Health workers in many parts of the world have a problem to successfully translate the term public health into their own languages bearing in mind the precise meaning of the concept. As for Serbia, the issue appeared to be solved 130 years ago, when B. Franklin's well-known sentence "Public health is public wealth" had been initially translated. The phrase used ("narodno zdravlje") was based on the already established German expression Volksgesundheit, but simultaneously in an optimal way reflected the idea of public health in Serbian. The adjective "narodno" has no exact equivalent in English, since it lacks socialistic connotation, as people's, informal one, as folk's, or too official one, as nation's. If any, the last option would still appear as the most adequate. In the recent past, the term public health services has been introduced, but enthusiasm for it fades away since this "innovation" demonstrates a lack of understanding of the whole broadness of the public health approach. Lawmakers in 2005 and 2009 replaced "narodno" with "javno" without consulting the academic community. This is an unfortunate move, because the only opposite of "javno" in Serbian is either "tajno" (secret) or "privatno" (private) and none of them fits well as a meaningful exclusion criterion. The author argues that the introduction of "javno" only serves a retrograde ideological prejudice and that tradition should be respected. PMID- 21626776 TI - [Advances in the study of histaminergic systems and sleep-wake regulation]. AB - Histaminergic neurons solely originate from the tuberomammillary nucleus (TMN) in the posterior hypothalamus and send widespread projections to the whole brain. Experiments in rats show that histamine release in the central nervous system is positively correlated with wakefulness and the histamine released is 4 times higher during wake episodes than during sleep episodes. Endogeneous prostaglandin E2 and orexin activate histaminergic neurons in the TMN to release histamine and promote wakefulness. Conversely, prostaglandin D2 and adenosine inhibit histamine release by increasing GABA release in the TMN to induce sleep. This paper reviews the effects and mechanisms of action of the histaminergic system on sleep-wake regulation, and briefly discusses the possibility of developing novel sedative hypnotics and wakefulness-promoting drugs related to the histaminergic system. PMID- 21626777 TI - [Research progress of the biological characteristics of IkappaB kinase and its inhibitors]. AB - The NF-kappaB pathway regulates the expression of over 150 target genes, e.g., cytokines, chemokines, leukocyte adhesion molecules and inducible effector enzymes. Consequently, it plays a crucial role in innate and adaptive immune responses, inflammatory response, stress responses, apoptosis and so on. IkappaB kinase (IKK) is the key of this pathway, and it owns a special structure which consists of catalytic subunit and regulatory subunit. Naturally, the activation of IKK needs the interaction of the two subunits and phosphorylation by its upstream kinases. Actually, there are two methods of activation of the NF-kappaB pathway, and both of the methods need the IKK complex. Given to the crucial role of IKK, researchers have isolated and synthesized amounts of IKK inhibitors, and these provide a great convenience to develop novel anti-inflammatory and anti tumor drugs. PMID- 21626778 TI - [Existing problems and strategies in liposome-mediated nucleic acid delivery]. AB - Liposome-mediated nucleic acid delivery has been a focus recently, but in the course of delivering nucleic acid, some hurdles seriously limit the nucleic acid exerting treatment effect. This review refers to a series of problems such as low blood stability, reticuloendothelial system absorption, the lower targeting of liposome and the restriction of endosomal escape which are suffered in liposome mediated nucleic acid delivery; and gives a detail introduction of strategies such as PEGylation, ligand modification, photochemical internalization, the application of degradation liposome and membrane-lytic peptide, to overcome those problems. PMID- 21626779 TI - [Effect of pravastatin on transportation of scutellarin in mouse liver and its mechanism]. AB - This study is to investigate the transportation of scutellarin in cell and live models and study on mechanism of absorption and transport of scutellarin in mouse liver. The concentration of scutellarin in plasma and liver from control and pretreated groups was determined by high performance liquid chromatography. The uptake of scutellarin was examined in control hepatocytes group, induced hepatocytes group and induced hepatocytes plus pravastatin group. Pravastatin can affect the pharmacokinetics of scutellarin in mouse: CL is decreased while AUC is increased. The scutellarin absorption of hepatocyte induced group was higher than that of control group, but was decreased in the group with pravastatin added. The research showed that there was potential drug interaction between pravastatin and scutellarin. The drugs may compete for oatp2 mediated transport pathway consisted in the uptake of scutellarin in liver. PMID- 21626780 TI - [Effect of combination of insulin and selenium on insulin signal transduction in cardiac muscle of STZ-induced diabetic rats]. AB - This study is to investigate the effect of low doses of insulin (1 u x kg(-1)) and selenium (180 microg x kg(-1)) in combination on general physiological parameters and insulin signal molecules in cardiac muscle of STZ-induced diabetic rats. The levels of blood glucose were estimated using One Touch SureStep Blood Glucose meter. HbA1c levels were estimated using microcolumn assay. TG and TC were estimated using enzymatic assay. The levels of PI3K and GLUT4 in cardiac muscle were examined by immunoblotting and immunohistochemistry. The result showed that insulin in combination with selenium could significantly lower blood glucose and blood lipid levels and markedly restored the PI3K and GLUT4 levels in cardiac muscle. It could be concluded that there was cooperation between insulin and selenium, and that treatment of diabetic rats with combined doses of insulin and selenium increased cardiac glucose uptake by upregulating the level of PI3K mediated GLUT4 in cardiac muscle, eventually ameliorating myocardial dysfunction. PMID- 21626781 TI - [Carrageenan oligosaccharides inhibit growth-factor binding and heparanase activity]. AB - This study is designed to investigate the anti-tumor and anti-angiogenesis mechanism of carrageenan oligosaccharides. The effects of carrageenan oligosaccharides on basic fibroblast growth factor (bFGF) induced cell proliferation, heparanase activity and bFGF binding ability were evaluated in human cervical cancer cells (HeLa) and human umbilical vein endothelial cells (HUVEC). Results indicate that, at rational concentrations, carrageenan oligosaccharides showed low cytotoxic effect. At relatively low concentrations (0.2-200 microg x mL(-1)), these oligosaccharides could competitively bind bFGF and inhibit bFGF induced cell proliferation. In these samples, oligo-lambda carrageenans (dp2-8) were the most potent bFGF antagonists. At concentration of 20 microg x mL(-1), their inhibitory ratio reached to 30%. The heparanase enzyme assay revealed that three kinds of carrageenan oligosaccharides showed different inhibitory activities to two cell lines. For HeLa cell, oligo-lambda-carrageenans showed highest inhibitory effect, but for HUVEC, oligo-kappa-carrageenans (dp9 17) were the best inhibitors. Current observations demonstrated that the biological activities of carrageenan oligosaccharides are closely related to the molecular weight, carbohydrate structure and the content and linking position of sulfur groups. Carrageenan oligosaccharides with high sulfate fraction, 2-8 units saccharide size and suitable molecular structure are able to achieve potent heparin sulfate-like compounds. PMID- 21626782 TI - [Modulation of Toll-like signal path of allergic asthma by CpG-ODNs from Bordetella pertussis]. AB - This study focused on prevention and treatment of acute and chronic asthma by oligonucleotides containing unmethylated CpG motifs (CpG-ODNs). Acute and chronic asthma models of mice were made by sensitizing and inhaling ovalbumin (OVA); The number of white blood cells (WBC) and eosnophils (EOS) in bronchoalveolar lavage fluid (BALF) was counted and the concentration of cytokines and vascular endothelial growth factor (VEGF) was examined in BALF by ELISA kit. After that, TLR-9 mRNA was detected in mice spleen cells by reverse transcription polymerase chain reaction (RT-PCR) and TLR-9 protein was determined in mice lung tissues by Western blotting. Compared with acute asthma models of mice, WBC in BALF decreased obviously in the groups of Bordetella pertussis, CpG-ODNs and seq A to seq I which were administrated by both of intragastric (ig) and intraperitoneal (ip) injection group, EOS decreased obviously in Bordetella pertussis, CpG+ and seq A to seq D ig groups, and in all ip administrating groups, although it was not effective in the groups of seq E to seq I. In chronic asthma models of mice, IFN-gamma increased ((1) control: 176.45 +/- 23.46 pg x mL(-1); (2) model: 174.11 +/- 22.71 pg x mL(-1); (3) CpG+ ip: 220.56 +/- 15.42 pg x mL(-1); (4) seq A ip: 225.23 +/- 21.60 pg x mL(-1)) and IL-4 decreased obviously (1) control: 66.91 +/- 5.81 pg x mL(-1); (2) model: 81.02 +/- 11.24 pg x mL(-1); (3) CpG+ ip: 63.99 +/- 6.09 pg x mL(-1); (4) seq A ip: 62.75 +/- 10.03 pg x mL(-1)) in the BALF of CpG+ and seq A ip group, although VEGF was not changed in this research. And also, TLR 9 mRNA in spleen cells (TLR-9/GAPDH: (1) control: 0.62 +/- 0.13; (2) model: 0.66 +/- 0.17; (3) CpG+ ip: 1.46 +/- 0.26; (4) seq A ip: 1.42 +/- 0.34) and TLR-9 protein in lung tissues (TLR-9/beta-actin: (1) control: 0.63 +/- 0.16; (2) model: 0.61 +/- 0.07; (3) CpG+ ip: 1.15 +/- 0.25; (4) seq A ip: 1.03 +/- 0.29) both increased in ip groups, but the change was not significant in ig group. The study confirms that CpG-ODNs and seq A could inhibit airway inflammation remarkably, this mechanism might be related with regulating Th1/Th2 balance and controlling the expression of TLR-9. PMID- 21626783 TI - [Design, synthesis, and biological activities of histone deacetylase inhibitors with diketo ester as zinc binding group]. AB - Histone deacetylases (HDACs) inhibition causes hyperacetylation of histones leading to growth arrest, differentiation and apoptosis of tumor cells, representing a new strategy in cancer therapy. Many of previously reported HDACs inhibitors are hydroxamic acid derivatives, which could chelate the zinc ion in the active site in a bidentate fashion. However, hydroxamic acids occasionally have produced problems such as poor pharmacokinetics, severe toxicity and low selectivity. Herein we describe the identification of a new series of non hydroxamate HDACs inhibitors bearing diketo ester moieties as zinc binding group. HDACs inhibition assay and antiproliferation assays in vitro against multiple cancer cell lines were used for evaluation. These compounds displayed low antiproliferative activity against solid tumor cells, while good antiproliferative activity against human leukemic monocyte lymphoma cell line U937. Compound CPUYS707 is the best with GI50 value of 0.31 micromol x L(-1) against U937 cells, which is more potent than SAHA and MS-275. HDACs inhibition activity of these compounds is lower than that expected, further evaluation is needed. PMID- 21626784 TI - [Design, synthesis and antiepileptic activity of 4-(2-acetoxybenzoylamino) butyrate derivatives]. AB - A series of 4-(2-acetoxybenzoylamino) butyrate derivatives were designed and synthesized. All of the novel 12 compounds (7a-7k) were synthesized from gamma aminobutyric acid (1) as starting material, and their structures were confirmed with IR, 1H NMR, EI-MS and elemental analysis. Preliminary pharmacological test in vitro showed that most of these title compounds possessed antiepileptic activity. Compounds 7i-7k displayed strong antiepileptic activity and are worth for further development. Compounds 4, 7d-7h showed moderate antiepileptic activity. The structure-activity relationship of 4-(2-acetoxybenzoylamino) butyrate derivatives is also discussed preliminarily. PMID- 21626785 TI - [Synthesis of ferulic acid derivatives and their inhibitory effect on platelet aggregation]. AB - Ferulic acid, an useful compound of Chinese traditional medicine, was used as leading compound. Six ferulic acid derivatives were designed and synthesized based on bioisosterism. Their structures were characterized by IR, 1H NMR, 13C NMR and mass spectra. In vivo experiment showed that ferulic acid derivatives had good inhibitory effects on adenosine diphosphate (ADP) induced platelet aggregation, which were significantly higher than that of Ozagrel. PMID- 21626786 TI - [Design, synthesis, and PPARalpha/gamma agonistic activity of novel tetrahydroisoquinoline derivatives]. AB - A series of tetrahydroisoquinoline derivatives were prepared and their peroxisome proliferator-activated receptor (PPAR) alpha/gamma agonistic activities were evaluated to obtain more potent PPAR agonist. All of them were new compounds, and their structures were confirmed by 1H NMR and HR-MS. Three compounds exhibited higher agonistic activities of PPARgamma than that of the comparison, six compounds exhibited higher agonistic activities of PPARalpha than that of the comparison, and compound 8a was discovered as a highly potent PPARalpha/gamma agonist that is much more active than that of WY14643 and rosiglitazone. The development of potent PPAR agonists may offer a new choice for the treatment of diabetes. PMID- 21626787 TI - [Chemical constituents from roots of Andrographis paniculata]. AB - To investigate the chemical constituents of the roots of Andrographis paniculata, 28 compounds were isolated and identified from the 80% ethanol extract. There are 20 flavonoids: 5, 5'-dihydroxy-7, 8, 2'-trimetroxyflavone (1), 5-hydroxy-7, 8, 2', 6'-tetramethoxyflavone (2), 5, 3'-dihydroxy-7, 8, 4'-trimethoxyflavone (3), 2'-hydroxy-5, 7, 8-trimethoxyflavone (4), 5-hydroxy-7, 8, 2', 3', 4' pentamethoxyflavone (6), wightin (7), 5, 2', 6'-trihydroxy-7-methoxyflavone 2'-O beta-D-glucopyranoside (8), 5, 7, 8, 2'-tetramethoxyflavone (10), 5-hydroxy-7, 8 dimethoxyflavanone (11), 5-hydroxy-7, 8-dimethoxyflavone (12), 5, 2'-dihydroxy-7, 8-dimethoxyflavone (13), 5-hydroxy-7, 8, 2', 5'-tetramethoxyflavone (14), 5 hydroxy-7, 8, 2', 3'-tetramethoxyflavone (15), 5-hydroxy-7, 8, 2' trimethoxyflavone (16), 5, 4'-dihydroxy-7, 8, 2', 3'-tetramethoxyflavone (17), dihydroneobaicalein (18), andrographidine A (19), andrographidine B (20), andrographidine C (21) and 5, 2'-dihydroxy-7, 8-dimethoxyflavone 2'-O-beta-D glucopyranoside (22); three diterpenoids: andrograpanin (23), neoandrographolide (24) and andrographolide (25); two phenylpropanoids: trans-cinnamic acid (26) and 4-hydroxy-2-methoxycinnamaldehyde (5); and oleanolic acid (9), beta-sitosterol (27) and beta-daucosterol (28). Compound 1 is a new flavone, compound 4 is a new natural product, compounds 2, 3 and 5 were isolated from the Androggraphis genus for the first time and compounds 6-9 were isolated from this plant for the first time. PMID- 21626788 TI - [Compatibility evaluation of Chinese medicines injections based on isothermal titration calorimetry]. AB - The paper is to report the establishment of a method for quickly evaluating compatibility of Chinese medicines injections. Isothermal titration calorimetry (ITC) was used to evaluate compatibility of Yiqifumai (YQFM) and vitamin C injection (Vc)/5% glucose injection (5% GS). The diversification of Gibbs free energy (deltaG), enthalpy (deltaH) and entropy (deltaS) were used to decide reaction types of colliquefaction procedures of different injections. The reactive profiles were used to determine signs and quantity of heat. And high performance liquid chromatography (HPLC) was used as a supportive method for ITC. Then, feasible binding sites were analyzed based on the information of spatial structures of major compositions. During the colliquefaction procedure of YQFM and Vc, [deltaH] > T[deltaS], so, the reaction is enthalpy-driving. And the reactive profile showed that a big deal of heat was given out during the procedure. Obviously, chemical reactions happened and the major compositions changed. On the other side, the reaction of YQFM mixed with 5% GS was entropy driving, because [deltaH] < T[deltaS]. The reactive profile showed there was a little heat gave out. So, non-chemical reactions happened, and the major compositions did not change. The conformity existed between the results of ITC and HPLC. ITC could be used to evaluate the compatibility of Chinese medicines injections because of the advantages of ITC, such as real time, fast, sensitive and having more parameters. PMID- 21626789 TI - [Pharmacokinetics study on characteristic ingredients of different-dose herbs of shaoyao-gancao decoction]. AB - The paper is to report the observation of pharmacokinetic changes of the characteristic ingredients in the combinations of different-dose herbs of Shaoyao Gancao decoction. After the establishment of HPLC analytical method of plasma effective constituents, rats were orally administered with different-dose herbs of Shaoyao-Gancao decoction. Blood samples at different times after administering these decoctions were collected, and then were analyzed by HPLC fingerprints technology. Pharmacokinetic parameters of characteristic peaks were analyzed by SPSS 15.0 software and DAS 2.0. At last, we looked for the correlation of those pharmacokinetic parameters and the dosage of Gancao. The best dose of Shaoyao Gancao decoction was at the ratio of 4 to 4, which was consistent with the dose commonly used in ancient times. The absorption of characteristic peaks from Shaoyao-Gancao decoction was related with the dosage of Gancao, and there existed interaction between each characteristic ingredients. There existed the right dose ratio of Shaoyao and Gancao to get the best effect. The absorptions of effective constitutents were mutual waxing and waning in order to increase biological effects together. It's demonstrated the compatibility connotation at a right dose ratio of Shaoyao-Gancao decoction through the angle of pharmacokinetics. PMID- 21626790 TI - [Study of pharmacokinetics of nicotine in local brain by using microdialysis and stable labeled isotope]. AB - The paper is to report the study of pharmacokinetics of transdermal administered nicotine in the brain of freely moving rat by using microdialysis with stable labeled isotope as internal standard. The pharmacokinetic behavior of nicotine in Sprague Dawley rat brain was investigated after intranasal administration (3.75 mg). Brain fluid samples were collected by intracerebral microdialysis with DL nicotine as internal standard. Concentrations of nicotine and DL-nicotine in the sample were measured by HPLC-MS/MS. Main pharmacokinetic parameters were calculated and analyzed by Das 2.0 pharmacokinetic software. The recovery of nicotine and the delivery of DL-nicotine were the same. The fate of absorption and distribution was two compartment model and the values of t1/2alpha was 170.31 min, t1/2beta was 263.30 min and the AUC(0-infinity) was 2.75 x 10(5) microg x L( 1) min separately. DL-nicotine can be used to calibrate the recovery of nicotine, and the new method of stable isotope microdialysis can be used to study the pharmacokinetics of freely moving rat. It will make sense for the treatment of addiction of tobacco and provide a new thought for the research of pharmacokinetics-pharmacodynamic combination. PMID- 21626791 TI - [Comparison of the characteristics of several polymer materials used in hydrophilic matrix tablets]. AB - Pure and drug hydrophilic matrix tablets were prepared by direct compression method with theophylline as a model drug. The characteristics of four hydrophilic matrix polymers, hydroxypropylmethylcellulose (HPMC), polyethylene oxide (PEO), sodium alginate (NaAlg) and xanthan gum (XG), were compared by investigating the water absorption, swelling, erosion and gel layer strength. The sequence of water absorption rate was XG >> NaAlg (H) > PEO > NaAlg (L) >> HPMC; The sequence of swelling index was XG >> PEO >> HPMC >> NaAlg; The sequence of erosion rate was NaAlg (L) > NaAlg (H) >> PEO80 > PEO200 > PEO300 > XG approximately PEO400 approximately K4M > K15M > PEO600 approximately K100M; The sequence of the gel layer strength was PEO > HPMC > XG >> NaAlg. For the PEO and HPMC matrix tablets, with the polymer molecular weight increased, the drug release mechanism was gradually transferred from mainly depending on the erosion to the diffusion; for SAL matrix tablets, the drug release mainly depends on erosion mechanism; and for XG matrix tablets, the drug release mainly depends on non-Fick diffusion mechanism. Comparison of the performance difference between the polymer materials will contribute to rational design and prediction of drug release behaviors from matrix tables and ultimately to achieve clinical needs. PMID- 21626792 TI - [Small interfering RNA delivery mediated by mPEG-PCL-g-PEI polymer nanoparticles]. AB - The aim of this paper is to report the synthesis of the mPEG-PCL-g-PEI copolymers as small interfering RNA (siRNA) delivery vector, and exploration of the siRNA delivery potential of mPEG-PCL-g-PEI in vitro. The diblock copolymers mPEG-PCL-OH was prepared through the ring-opening polymerization. Then, the hydroxyl terminal (-OH) of mPEG-PCL-OH was chemically converted into the carboxy (-COOH) and N hydroxysuccinimide (NHS) in turn to prepare mPEG-PCL-NHS. The branched PEI was reacted with mPEG-PCL-NHS to synthesize the ternary copolymers mPEG-PCL-g-PEI. The structure of mPEG-PCL-g-PEI copolymers was characterized with Fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR) and gel permeation chromatography (GPC). The mPEG-PCL-g-PEI/siRNA nanoparticles were prepared by complex coacervation, and the nanoparticles size and zeta potential were determined, separately. The cytotoxicities of mPEG-PCL-g-PEI/siRNA nanoparticles and PEI/siRNA nanoparticles were compared through cells MTT assays in vitro. The inhibition efficiencies of firefly luciferase gene expression by mPEG-PCL-g-PEI/ siRNA nanoparticle at various N/P ratios were investigated through cell transfection in vitro. The experimental results suggested that the ternary (mPEG5k-PCL(1.2k))1.4-g-PEI(10k) copolymers were successfully synthesized. (mPEG(5k)-PCL(1.2k))1.4-g-PEI(10k) could condense siRNA into nanoparticles (50-200 nm) with positive zeta potential. MTT assay results showed that the cytotoxicity of (mPEG(5k)-PCL(1.2k))1.4-g-PEI(10k)/siRNA nanoparticles was significantly lower than that of PEI(10k)/siRNA nanoparticles (P < 0.05). The expression of firefly luciferase gene could be significantly down-regulated at a range of N/P ratio from 50 to 150 (P < 0.01), and maximally inhibited at the N/P ratio of 125. The mPEG-PCL-g-PEI polymers could delivery siRNA into cells to inhibit the expression of target gene with very low cytotoxicity, which suggested that mPEG-PCL-g-PEI could serve as a new type of siRNA delivery vector. PMID- 21626793 TI - [Tissue distribution of free anthraquinones in SD rats after orally administered extracts from raw and prepared rhubarbs]. AB - Rhubarb anthraquinone derivatives (AQs) have been documented to have both therapeutic and toxic effect on liver and kidney, leading to a complex puzzle to assess their benefits and risks. In this study, the tissue distributions of AQs in SD rats after orally administrated extracts of raw and prepared rhubarb were examined whether they undergo different uptake. The total rhubarb extract (14.49 g x kg(-1) of body weight per day od, counted on the quantity of crude material) was administrated orally for 12 weeks. The concentrations of the AQs in different tissues were quantified by liquid chromatography-tandem mass spectrometry (LC MS/MS). The five major AQs, aloe-emodin, rhein, emodin, chrysophynol and physcion, could all be detected in the liver, kidney and spleen, while only rhein, aloe-emodin and emodin reached the quantitative limit. The tissue concentrations of AQs in raw rhubarb group were higher than that in steamed rhubarb group with rhein > emodin > aloe-emodin in the same tissue. On the whole, the tissue distribution of rhein was higher than that of emodin and aloe-emodin in liver, spleen and kidney. AQs could not be detected in those tissues after drug withdraw for 4 weeks, which suggested scarcely any accumulative toxicity of rhubarb. The result indicated that raw rhubarb had more tissue toxicity than steamed rhubarb and rhein may be one of the major poisonous ingredients. The results were concordant with the traditional Chinese medicine theory of toxicity attenuating effect of processing. PMID- 21626794 TI - [Regulation of Vitreoscilla hemoglobin on biosynthesis of astragaloside IV]. AB - In the present study, the regulation of Vitreoscilla hemoglobin (VHb) on astragaloside IV biosynthesis was investigated. An intermediate expression vector consisting of the CaMV35S promoter fused to the vgb and nopaline synthase terminator was transferred into Astragalus membranaceus via Agrobacterium rhizogenes. The transgenic hairy roots were confirmed by PCR amplification and Southern blot hybridization. The expression of vgb in transgenic hairy roots was confirmed by RT-PCR. After 15 days cultivation, the dry weight and growth rate of transgenic hairy roots were higher than that of the non-transgenic hairy root. ELSD-HPLC analysis showed that astragaloside IV content of transgenic hairy roots was 5 to 6 times of non-transgenic hairy root control and 10 to 12 times of Radix Astragali from Shanxi Province. These results suggested that the expression of vgb promoted the growth of transgenic hairy roots, and increased the content of astragaloside IV. PMID- 21626795 TI - [Comparative analysis of antioxidant activity of nebivolol in patients with chronic heart failure with and without concomitant type 2 diabetes]. AB - Consistent neurohormonal activation of sympatho-adrenal system in patients with chronic heart failure (CHF) and hyperglycemia contributes to development of oxidative stress--one of the most important pathogenetic mechanisms of endothelial dysfunction. PURPOSE: To study the impact of nebivolol concerning modification of clinical and hemodynamic indicators and parameters of oxidative stress in patients with CHF and with or without concomitant diabetes mellitus type 2 (DM2). MATERIAL: Nebivolol was used in complex therapy of CHF in 82 patients, suffering from NYHA class I - III CHF (EF < 50%) of ischemic genesis with or without comorbid DM2, average age 63.2 +/- 8.2 years. RESULTS: After 8 months of therapy significant improvement of clinical status was observed in both groups, tolerance to physical activity increased (significant reduction of average class of CHF in the group with DM2 from 2.5 +/- 0.58 to 2.125 +/- 0.71, p = 0.001, and in the second group from 2.3 +/- 0.5 to 1.9 +/- 0.4, p = 0.01). We also noted in both groups increase of plasma oxidative resistance (reduction of intensity of fast flash in lipid peroxidation h from 7 to 6 mm, p = 0.016, and from 8 to 6 mm, p = 0.03, respectively) and increase of antioxidant plasma protection (increase of SH-groups from 154.19 to 182.4 mmol/1, p = 0.00035, and from 176 to 205, p = 0.004, respectively). CONCLUSION: Nebivolol is a modern neurohormonal modulator, which contributes to reverse evolution of oxidative changes in patients with CHF and hyperglycemia. PMID- 21626796 TI - [The statistical analysis of prevalence of anemia in patients with chronic heart failure]. AB - Development of anemia in patients with congestive heart failure (CHF) is associated with worsening of disease what has been shown in numerous trials. According to various data prevalence of the syndrome varies depending on chosen populations of patients and criteria of anemia. We observed 1122 patients (mean age 71.5 +/- 0.56 years) with CHF NYHA class II-IV (average 3.08 +/- 0.022). Anemia was found in 317 patients (28.3%); 59.3% of these patients were women and 40.7% - men. There were significantly more subjects with NYHA class IV CHF among patients with anemia (p < 0.001). Hospital mortality was 14.8 and 13.4% among patients with anemia and normal hemoglobin, respectively (p = 0.038). Analysis of mortality structure showed that among patients with anemia compared with those without there were fewer deaths due to pulmonary embolism (27.6 vs. 43.5%, p = 0.044) and more deaths due to decompensation of CHF (60.3 vs. 41.7%, p = 0.022). So this study demonstrated high rate of anemia among hospitalized patients with CHF and its association with higher NYHA class and worse prognosis. PMID- 21626797 TI - [High level of lipoprotein (a) as a predictor of poor long-term prognosis after coronary artery bypass surgery]. AB - Study aim was to investigate the association of lipoprotein (a) [Lp(a)] level with the development of cardiovascular complications in long-term follow-up period after coronary artery bypass grafting (CABG). Patients with chronic ischemic heart disease (IHD) (n = 361, 88% men, mean age 55 +/- 9 years) who had had CABG were included in the study. Before surgery we assessed presence of classical risk factors, left ventricular ejection fraction, concentrations of lipids and Lp(a) in blood serum. During follow-up (from 1 to 140, mean 66 +/- 34 months) we registered cardiac deaths, nonfatal myocardial infarctions (MI), strokes, repeat procedures of revascularization, and hospitalizations due to relapse or progression of angina pectoris. Information on prognosis was obtained from 263 patients. In 109 of them we registered 142 serious events including cardiac death n = 20 (14%), nonfatal MI n = 14 (10%), myocardial revascularization (n = 35), 29 (20%) with stenting), repeat CABG n = 6 (4%), hospitalization due to angina pectoris n = 53 (37%), stroke n = 4 (3%), noncardiac outcome n = 16 (10%). In subjects with hyperlipidemia (a) [HLp(a) - Lp(a) > 30 mg/l] survival after CABG was lower (log rank p < 0.001): 11 of 93 (11.3%) and 9 of 170 (5.2%) patients died among those with Lp(a) > 30 and < 30 mg/I, respectively. Relative risk (RR) of any cardiovascular complication was 3.24 (95% confidence interval [CI] 2.18 to 4.83, p < 0.001), of death - 2.89 (95% CI 1.31 to 6.35, p < 0.01), and of MI A 1.01 (95% CI 1.00 to 1.02; p = 0.02). RR of development of MI and cardiac death in patients with HLp(a) in 5 years was 2.61 (95% CI 1.11 to 5.74; p = 0.02), in 10 years - 2.95 (95% CI 1.50 to 5.79; p < 0.001). In patients with chronic IHD high level of Lp(a) can serve as independent predictor of unfavorable events including death and nonfatal MI during 10 years after CABG. PMID- 21626798 TI - [Prognostic value of aortic stenosis in patients after acute coronary syndrome]. AB - With the aim to assess prevalence of aortic stenosis (AS) and prognostic value of its detection among survivors of acute coronary syndrome (ACS) we examined 851 patients included into multicenter prospective study of risk factors of serious vascular events and death after acute coronary syndrome. The patients were enrolled into the study in stable condition on 10th day after onset of myocardial infarction (MI) or unstable angina (UA). Examination involved medical history, laboratory tests and echocardiography. Afterwards all cases of death and serious vascular events were registered. Severity of AS was specified by maximal aortic flow rate: 1st degree > 2.5, 2nd degree 3.0-4.0, 3rd degree > 4.0 m/s. AS was detected in 16 patients (1.9%). AS severity was 1st, 2nd and 3rd degree in 9, 4 and 3 patients, respectively. Patients with AS were significantly older (77.4 vs. 61.3 years, p < 0.001), more often had history of chronic heart failure (CHF) (81.3 vs. 53.2%, p = 0.021) and lowered renal function (66.7 vs. 34.0%, p < 0.041). At multifactorial analysis independent prognostic value in relation to development of serious events showed age > 75 years (OR 1,395 [1.023-1.902], p = 0.036), history of CHF (1.319 [1.015-1.713], p = 0.038), history of MI (1.692 [1.320-2.170], p < 0.001), left ventricular diastolic dimention (1.023 [1.005 1.041], p = 0.012), left atrial diameter (1.024 [1.001-1.047], p = 0.037) and presence of AS (3.211 [1.742-.,916], p < 0.001). Prevalence of preexisting AS among patients who have had MI/UA is 1.9% what is similar to data of European Heart Survey ACS-II (1.8%). Presence of AS of any severity in a survivor of ACS worsens prognosis independently of other known risk factors. PMID- 21626799 TI - [Dynamics of morphological structures of platelets in patients with ischemic heart disease in dependence on blood levels of fatty acids]. AB - Levels of fatty acids in platelets of ischemic heart disease (IHD) patients (n = 39) has been compared with those in healthy subjects (n = 12). Increased content of arachidonic acid and thromboxane A2 in platelets of IHD patients forms thrombogenic picture of IHD. High level of fibrinogen and decrease in heparin and antithrombin III in IHD patients facilitates formation of blood clots. Morphological examination of platelets in IHD patients has demonstrated an increase of levels of discocytes and spherocytes as well as appearance of small and large platelet-erythrocyte aggregates promoting blood slot formation. PMID- 21626800 TI - [Monitoring of secondary prevention of ischemic heart disease in Russia and European countries: results of international multicenter study EUROASPIRE III]. PMID- 21626801 TI - [Own and parental predictors of high level of low density lipoprotein cholesterol at children of persons with "premature" coronary heart disease]. AB - AIM: To elucidate associations between high low density lipoprotein cholesterol (LDL-C) in children of persons with premature coronary heart disease (PCHD) and their own and parental characteristics. METHODS: We examined members of 282 families: probands (n = 262, 70.1% men) with PCHD (onset before 55 [men] or 60 [women] years), their spouses (n = 192, 19.2% men), and children of probands (n = 372, 55.8% men) aged 5-38 years. We defined high LDL-C in children aged 5-17 years as a90 percentile (Lipid Research Clinics Program Prevalence Study), in children aged 18-38 years - > or = 3.36 mmol/l. Factors related to children's high LDL were selected by logistic regression with sex and age adjustment. RESULTS: High LDL-C was found in 40/155 (25.8%) children aged 5-17 years. It was independently related to non-smoking of proband's spouse, higher own body mass index (BMI), LDL-C of proband and his spouse, lower heart rate of proband. High LDL-C was revealed in 85/217 (39.2%) children aged 18-38 years. It was independently related to metabolic syndrome of parent-nonproband, higher level of LDL-C of both parents, lower height of proband. INTERPRETATION: Most associations of high LDLAC of offsprings of patients with PCHD including negative relation to height and heart rate of proband are plausible. Association with parental nonsmoking can be result of analysis of unreliable data from small group but also reflect better economical conditions and more atherogenic nutrition of nonsmokers in Russia. PMID- 21626802 TI - [Preventive research in action]. PMID- 21626803 TI - [Percutaneous coronary interventions in patients with acute myocardial infarction after unsuccessful thrombolysis]. AB - In 176 patients with acute myocardial infarction admitted to N.V. Sklifosofsky institute of urgent aid in 2003-20007 we compared efficacy of 3 strategies of treatment after unsuccessful thrombolytic therapy (TLT): percutaneous coronary intervention (PCI) during first 24 hours (n = 30), PCI on days 2 or 3 (n = 38); conservative treatment (n = 108). The data obtained show that it is expedient to consider absence of 50% reduction of STAsegment elevations in 90 min after start of TLT as indication to urgent late PCI when possibilities for immediate intervention after unsuccessful thrombolysis are lacking. Alternative reperfusion is the only type of effective treatment of patients with failed pharmacological reperfusion. Necessity to perform PCI during first 12 hours after unsuccessful TLT does not exclude possibility of its later fulfillment in acute period of myocardial infarction. Efficacy of the latter is comparable with success rate of rescue PCI. The use of both invasive strategies has allowed to lessen rate of complications and prevent lethal outcomes. Success of late urgent interventions in acute period of infarction after failed thrombolysis opens possibilities for their active use in patients transferred from other hospitals. PMID- 21626804 TI - [First experience of clinical use of new class III antiarrhythmic agent niferidil in patients with persistent atrial fibrillation and flutter]. AB - The aim of the study was to evaluate the efficacy and safety of administered intravenously niferidil in doses 10, 20 and 30 mkg per kg in patients with persistent atrial fibrillation (AF) and flutter (AFL) for pharmacological cardioversion. The study included 30 patients (22 male) with persistent AF (n = 28) and AFL (n = 2) without structural heart diseases with median arrhythmia duration 6.1 +/- 4.8 months (2 weeks to 24 months). Niferidil was administered as 3 bolus injections (10 mkg per kg each) performed with the interval of 15 minutes. Antiarrhythmic efficacy of niferidil in dose of 10 mkg per kg was 60%, in dose of 20 mkg per kg it was 70%, and in dose of 30 mkg per kg reached 90% prespectively. The part of the patients, in whom QTc prolongation exceeded potentionally dangerous value of 500 mc, was 22.2% (6 of 27). None of the patients developed proarrhythmic side effect as torsade de pointes. PMID- 21626805 TI - [Antitachycardia pacing in patients with implanted cardioverter defibrillators]. AB - Efficacy and safety of the use of antitachycardia pacing (ATP) function in 54 patients (age 18-54, mean age 62.2 +/- 11.5 years, 6 women) with cardioverter defibrillators implanted for prevention of sudden death. Duration of follow-up was 6-64 months (mean 27.3 +/- 8.6 months). Ischemic heart disease (IHD) was etiopathogenetic basis for development of ventricular rhythm disturbances in 38 (70.4%) patients. Arrhythmogenic right ventricular dysplasia (ARVD) was diagnosed in 8 (14.8%), hypertrophic cardiomyopathy--in 3, dilated cardiomyopathy--in 2, the Brugada syndrome--in 2 patients, and long QT syndrome--in 1 patient. Three regimens of ATP were studied--burst, ramp, ramp(+)--which were activated in 22 patients (17 with IHD and 5 with ARVD). We registered 776 episodes of ATP occurring because of ventricular tachycardia (VT) in 20 of 22 patients with activated ATP. Overall efficacy of burst ATP was 49.1%; VT was not terminated or was accelerated in 43.2 and 7.6% of cases, respectively. Ramp ATP was effective in 44.3, ineffective--in 33.9, and caused VT acceleration in 21.8% of cases, what called forth fulfillment as a next step of shock therapy (cardioversion or defibrillation). ATP led to successful termination of VT in 67.5% of patients with IHD and to acceleration of arrhythmia--in 6% of episodes. Most effective type was ramp ATP which effectively terminated VT in 76.3% of cases, while efficacy of burst ATP was 61.4% (p < 0.01). In patients with ARVD total efficacy of ATP was 25%, acceleration of VT during ATP was observed in 24% of cases; optimal was burst ATP, which efficacy was 33.9%--significantly higher than efficacy of ramp and ramp+ ATP (19.4 and 24.4%, respectively, p < 0.005). PMID- 21626806 TI - [Antihypertensive and nephroprotective effectiveness of AT1-angiotensin II receptor blocker valsartan in average therapeutic and maximal daily dose in patients with arterial hypertension and chronic kidney disease]. AB - The results of the study which show efficiency of an antihypertensive and nephroprotective angiotensin II receptor blocker valsartan in patients with arterial hypertension of and chronic kidney disease are presented. Dose dependence of nephroprotective and antihypertensive effects of valsartan is demonstrated as well as high safety profile of the drug in average and maximal therapeutic doses. PMID- 21626807 TI - [Diastolic dysfunction]. PMID- 21626808 TI - [Achievements and problems of modern trials of antihypertensive drugs]. AB - Most important value of lowering of substantially elevated arterial pressure (AP) for improvement of outcomes in patients with arterial hypertension (AH) was convincingly confirmed by large truly placebo controlled randomized clinical trials (RCT) with the use of mainly diuretics, and/or beta-adrenoblockers in the 60-80ths. Later comparative RCT confirmed equal antihypertensive efficacy of 5 main drug classes relative to AP level in brachial artery. In this review we discuss merit of auxiliary class-specific properties of antihypertensive agents potentially affecting prognosis besides AP lowering. We also discuss problems related to decline of significance of quantitative criteria of AH and consideration of AP level in general context of cardiovascular risk; problems of external validity of RCT; extrapolation of RCT results obtained in patients with complicated AH and very high cardiovascular risk on young patients with uncomplicated AH; significance of hard and surrogate end points. PMID- 21626809 TI - [Pulmonary arterial hypertension: changing approaches to management]. AB - The review is devoted to different aspects of pulmonary arterial hypertension (PAH); new classification of PAH is published in 2010. There are idiopathic PAH and PAH associated with other diseases. Current guidelines recommend to treat PAH only after the verification of diagnosis with right heart catheterization and acute tests with vasodilators. Patients-reactors should be treated with calcium antagonists. The following drugs related to one of three categories should be used in PAH: (1) prostanoids (epoprostenol, iloprost et al.); (2) blockers of endothelin receptors (bosentan, ambrisentan, sitaxsentan); (3) phosphodiesterase 5 type inhibitors (sildenafil, tadalafil et al.) In majority of cases the combined treatment is used, usually the combination of bosentan and sildenafil is used. PMID- 21626810 TI - [On two variants of viral injury of the myocardium]. PMID- 21626811 TI - [Combination of perindopril and amlodipine as optimal basis for realization of desirable clinical effects in the treatment of arterial hypertension with fixed drug combination]. PMID- 21626812 TI - [Risk factors and prediction of poststroke dementia]. AB - To study the status of risk factors for poststroke dementia, 128 patients, aged 50-79 years, with poststroke dementia have been examined. A control group included 125 patients with moderate cognitive impairment after stroke. Coronary heart disease was a significant factor at the age from 50 to 59; coronary heart disease, diabetes mellitus, overweight and hyperlipidemia were significant risk factors at the age from 60 to 69 and diabetes mellitus, overweight and hyperlipidemia were significant risk factors at the age from 70 to 79. The neuroimaging study revealed that the development of dementia in patients with moderate cognitive impairment was related to the localization of focal poststroke changes in the left hemisphere, frontal and temporal-occipital areas, frontal white matter, thalamus, basal ganglia as well as to the prevalence and severity of subcortical leukoareosis in the frontal areas, basal ganglia and thalamus. An individual prediction model for poststroke dementia in patients with moderate cognitive impairment has been worked out. PMID- 21626813 TI - [Dynamics of clinical and laboratory parameters in patients with severe ischemic stroke of hemispheric localization and different variants of nutrition support]. AB - We studied an effect of different nutrition algorithms on the outcome of neurological symptoms and laboratory data in patients with ischemic stroke. The active nutrition support included in the basic therapy from the beginning of the disease has a positive effect on the patient's state. The mixed artificial nutrition (enteral + intravenous) was more effective compared to the intravenous or enteral nutrition alone in improving the clinical state, shortening the duration of staying in intensive care wards and complete compensation of disturbances of protein and energy metabolism during the first day of disease. PMID- 21626814 TI - [Antithrombotic therapy in the secondary prevention of stroke]. AB - The review is devoted to the secondary prevention of ischemic events after ischemic stroke. Recommendations for antithrombotic therapy and prolonged indirect anticoagulant therapy with INR monitoring based on evidence-based medicine of the secondary prevention of ischemic stroke and TIA are presented. Patients who do not need anticoagulants should receive antiplatelet therapy. Due to the increasing number of patients resistant to antiaggregant therapy, the rapid and adequate evaluation of platelet function is needed. Current laboratory diagnostic methods have some shortcomings and do not meet criteria of evidence based medicine. It is necessary to improve the laboratory diagnosis with the elaboration of reliable standardized methods. PMID- 21626815 TI - [The use of akatinol memantine in the residual phase of stroke]. AB - Fifty patients who had had left-hemisphere stroke 1-12 months before the inclusion in the study were examined. All patients underwent the 90-day course of neurorehabilitation. Forty patients of the main group received akatinol memantine as an add-on drug during 90 days in the initial dosage 5-10 mg/day with the gradual increase to 20 mg/day (10 mg twice a day); the control group included 10 patients. Comparative efficacy of neurorehabilitation was assessed 1,5 and 3 months after treatment with clinical scales NIHSS and Barthel, testing of higher mental functions with 10-score original digital scale (V.M. Shklovsky et al), neurophysiological study (electroencephalography and constant brain potentials). The distinct effect of akatinol memantine on the restoration of higher mental functions and, to a less extent, on the movement deficit was seen. The dynamic neurophysiological study revealed that the addition of akatinol memantine to the neurorehabilitation course activates reserve possibilities of the left hemisphere in small or moderate poststroke lesions. The trend toward the association between the efficacy and the time since stroke onset and the intensity of rehabilitation measures was found. PMID- 21626816 TI - [The effect of cerebrolysin in dosage 50 ml on the volume of lesion in ischemic stroke]. AB - The purpose of this randomized, double-blind, placebo-controlled study was to assess safety and efficacy of cerebrolysin used in dosage 50 ml in acute ischemic stroke. Forty-seven patients with ischemic stroke, aged 45-85 years, who were admitted to a clinical unit within the first 12 h after stroke onset were included in the study. A quantitative time-related MRI analysis of the dynamics of neurological deficit revealed the more rapid decrease of stroke volume to the 28th day in the group treated with cerebrolysin (45.4% versus 43.6% in the placebo-group (p < 0.05)). No side-effects of treatment with cerebrolysin was found. The results of this prospective, randomized, placebo-controlled study suggest the positive effect of cerebrolysin on the dynamics of volume lesion in patients with ischemic stroke. PMID- 21626817 TI - [Efficacy of phenotropil in the rehabilitation of stroke patients]. AB - We studied 400 patients with ischemic stroke, including 200 patients (the main group), who received 3 courses of phenotropil in the complex rehabilitation treatment in dosage 400 mg/day during 1 year after stroke. The extent of restoration of neurologic functions and daily living activities was assessed with Barthel, Lindmark, Scandinavian scales and Merton and Sutton scale. The restoration of neurologic and daily living activities was significantly (p < 0.0001) better in the main group compared to the control one. PMID- 21626818 TI - [Disturbances of lipid metabolism in the period of age androgen deficit in middle aged and elderly men with ischemic stroke]. AB - Blood concentrations of total cholesterol, beta-lipoproteins, triglycerides and testosterone were studied in 73 men (mean age 65.9 +/- 8.6 years) with acute ischemic stroke. The concentrations of testosterone and triglycerides in men decrease with age and those of cholesterol and beta-lipoproteins do not depend on the age. In middle-aged patients, the negative correlations were found between concentrations of testosterone and beta-lipoproteins (r = -0.404; p < 0.05); testosterone and cholesterol (r = -0.273; p < 0.001). In elderly patients, there were negative correlations between concentrations of testosterone and triglycerides (r = -0.348; p < 0.05). In middle-aged and elderly patients, concentrations of triglycerides and testosterone were negatively correlated (r = 0.478; p < 0.001). The lower blood testosterone level was associated with the higher risk of the development of atherosclerotic lesions of major arteries that allowed to estimate risks of ischemic stroke in middle-aged and elderly men. PMID- 21626819 TI - [Treatment of patients with movement and cognitive disorders in the residual period of stroke]. AB - The efficacy and safety of axamon was studied in a trial with parallel groups in patients, aged 65-88 years, with movement and cognitive disorders in the residual period of stroke. Twenty-five patients received basic therapy and 25 patients were treated with axamon as an add-on drug in dosage of 20 mg 3 times a day during 12 weeks. Along with the neurological examination, a battery of neuropsychological scales and tests was used. The reduction of cognitive and movement deficits was observed. Axamon promotes the optimization of restoration potential of patients in the residual period of stroke. PMID- 21626820 TI - [Correlation between quality of life and social/demographic factors in patients with stroke]. AB - The population of Kaunas was studied in 2004-2006. The main group included 508 patients with first-ever stroke and the control group included 508 age- and sex matched people randomly selected from the population. The SF-12 quality of life questionnaire in the domains of physical and mental health was used. The significant inverse correlation (p < 0.05) between the physical health and the age was found both in patients (r = -0.34) and controls (r = -0.64). The scores of physical health were higher in men (p < 0.001) as compared to women only in the control group. The physical health of Livonian patients was better (p < 0.008) than that of patients of other ethnic groups. People living alone scored their physical health lower as in the control (p < 0.01) as well in main (p < 0.008) groups. The physical health of working people was better (p < 0.005) in both groups. In both groups, subjects with higher education and those not living alone reported themselves in better health regardless of sex compared to people with lower education and/or those living alone and/or unemployed. PMID- 21626821 TI - [Influence of the neural stem cell transplantation on the restoration of CNS functions in rats with cortical stroke]. AB - Ischemic stroke was modeled in white pedigreeless rats by the superficial blood vessel devascularization in the sensorimotor cortex. The preparations of neural progenitors--rat embryonic neural stem cells (rENSC) and human olfactory epithelium-derived neural stem cells (hOENSC) and differentiated fibroblasts ("cell control") were transplanted at the perimeter of the devascularized region. These cells marked with vital tracer stayed alive in the brain parenchyma for at least 16 days. The monitoring of contralateral forepaw motor deficit during 8 weeks demonstrated that only rats with rENSC transplantation had the stable and significant improvement of performance in cylinder test and swimming test (forepaw inhibition test) in comparison to "cell controls" and rats without cell transplantation. The maximal difference in the relative values (the efficacy) was 25% to the end of the experiment. There was no difference in the indicators of vibrissae-elicited forelimb placing test between experimental groups. The methodological approach used makes it possible to broaden the study of mechanisms of neural stem cells' therapeutic effect in stroke. PMID- 21626822 TI - [Obesity: Progress in diagnosis and treatment; A disease entity peculiar to Japan, himansho, obesity as a disease]. PMID- 21626823 TI - [Obesity: Progress in diagnosis and treatment; Topics, I. Diagnosis and related examinations; 1. Obesity and obesity disease]. PMID- 21626824 TI - [Obesity: Progress in diagnosis and treatment; Topics, I. Diagnosis and related examinations; 2. Specific health check-up and health counseling]. PMID- 21626825 TI - [Obesity: Progress in diagnosis and treatment topics, I. Diagnosis and related examinations; 3. Obesity and adipocytokine]. PMID- 21626826 TI - [Obesity: Progress in diagnosis and treatment; Topics, II. Progress in treatment; 1. Behavioral therapy for obesity]. PMID- 21626827 TI - [Obesity: Progress in diagnosis and treatment; Topics, II. Progress in treatment; 2. Drug strategies for the treatment of obesity--clinical application of gut brain peptides]. PMID- 21626828 TI - [Obesity: Progress in diagnosis and treatment; Topics, II. Progress in treatment: 3. Current status of bariatric surgery]. PMID- 21626829 TI - [Obesity: Progress in diagnosis and treatment; Topics, III. Obesity and its complications; 1. Obesity and diabetes]. PMID- 21626830 TI - [Obesity: Progress in diagnosis and treatment; Topics, III. Obesity and its complications: 2. Obesity and hypertension]. PMID- 21626831 TI - [Obesity: Progress in diagnosis and treatment; Topics, III. Obesity and its complications; 3. Obesity and dyslipidemia]. PMID- 21626832 TI - [Obesity: Progress in diagnosis and treatment; Topics, III. Obesity and its complications; 4. Obesity and its vascular complication]. PMID- 21626833 TI - [Obesity: Progress in diagnosis and treatment; Topics, III. Obesity and its complications; 5. Obesity and sleep disorders]. PMID- 21626834 TI - [Obesity: Progress in diagnosis and treatment: Topics, III. Obesity and its complications: 6. Cancer associated with obesity]. PMID- 21626835 TI - [Obesity: Progress in diagnosis and treatment topics, IV. Recent topics; 1. Ectopic fat disposition and lipotoxicity]. PMID- 21626836 TI - [Obesity: Progress in diagnosis and treatment; Topics, IV. Recent topics; 2. Obesity and inflammation]. PMID- 21626837 TI - [Obesity: Progress in diagnosis and treatment; Topics, IV. Recent topics: 3. Obesity and new secretory factors; 1) Vaspin]. PMID- 21626838 TI - [Obesity: Progress in diagnosis and treatment; Topics, IV. Recent topics; 3. Obesity and new secretory factors; 2) Sfrp 5 acts as a novel adipocytokine that affects metabolic function]. PMID- 21626839 TI - [Discussion meeting on the management of metabolic syndrome for the prevention of cardiovascular event]. PMID- 21626840 TI - [Case report; the clinicopathological report of a 68-year-old woman of pneumococcal meningitis with diffuse stenosis of the cerebral arteries]. PMID- 21626841 TI - [Case report; a case of familial Mediterranean fever diagnosed in adult]. PMID- 21626842 TI - [Case report: IgG4-related hypophysitis presenting with secondary adrenal insufficiency and central diabetes insipidus in a type 1 diabetes patient]. PMID- 21626843 TI - [Case report; mycotic aneurysm caused by Listeria monocytogenes]. PMID- 21626844 TI - [Case report; autoimmune polyendocrine syndrome type3 with thyrotoxic crisis and diabetic coma]. PMID- 21626845 TI - [Case report; a case of MALT lymphoma related cryoglobulinemic MPGN responded to combined chemotherapy with rituximab]. PMID- 21626846 TI - [The cutting-edge of medicine; the cutting edge of pulmonary nontuberculous mycobacterial diseases]. PMID- 21626847 TI - [The cutting-edge of medicine; waon therapy for cardiovascular disease]. PMID- 21626848 TI - [The cutting-edge of medicine; cerebrospinal fluid hypovolemia]. PMID- 21626849 TI - [Series: Knowledge of emergency required for internist; shock]. PMID- 21626850 TI - [Report from the 7th Tohoku Chapter Educational Seminar: A case of 57-year old woman with disturbed consciousness and difficulty in body movement]. PMID- 21626851 TI - [Series: Let's think-clinical quiz (question); A 80-year-old woman with ST segment elevation following sudden onset respiratory failure after thymectomy in myasthenia gravis]. PMID- 21626852 TI - [Series: Clinical study from Japan and its reflections: introduction and summary of results: the ESTABLISH study]. PMID- 21626853 TI - [Series: Diagnosis at a glance]. PMID- 21626854 TI - [Series: For attending physicians; professionalism; the true nature of professionalism: understanding altruism and social contract]. PMID- 21626856 TI - [How far has our understanding of mechanisms of general anesthesia advanced?: preface and comments]. AB - Although the great advance has been made in clinical anesthesia practice, the fundamental mechanisms of anesthetic action still remain to be an unsolved mystery. The early lipid membrane theory based on Meyer and Overton's law was taken over by the proteo-centric view of mechanism. Studies at the molecular and cellular level have shown that anesthetics act on a wide rage of functional proteins, including ligand-gated ion channels (GABA, glycine, NMDA receptors), two pore domain K channels and other ion channels. The effects on the individual channels, however, differ among various types of anesthetics. Elucidating how anesthetics work on the neuronal pathways is important to find the link between the molecular studies and in vivo action of anesthetics. Anesthesia disrupts the linkages between cortical and thalamic neurons and those among the cortical neurons, as well as depression and activation of the arousal and sleep nuclei, respectively. Functional brain imaging has revealed the global effects of general anesthetics on the human brain. Taken together, the disruption of thalamocortical connectivity and the integrative properties of the cerebral cortex might be an essential common feature of anesthetic action. PMID- 21626857 TI - [Roles of glutamatergic and GABAergic nervous system in hypnotic and analgesic actions of general anesthetics]. AB - General anesthetic-induced unresponsiveness covers a spectrum of different behavioral components, namely, (1) amnesia, (2) sedation/hypnosis, (3) analgesia, and (4) immobility. At the molecular and cellular level, anesthetic drugs have been shown to have effects on a wide rage of putative targets, such as ligand gated ion channels (GABA, glycine, NMDA receptors), other ion channels (K+, Na+, Ca2+), and other intracellular functions. This mini-review summarizes recent topics in this research field focusing on NMDA and GABA receptors. Although ketamine blocks NMDA receptors as an open channel blocker, it has been recently shown that ketamine inhibits hyperpolarization-activated cationic currents (J Neurosci 2009) and also enhances GABA-induced currents in alpha6 GABA receptors (J Neurosci 2008). In addition, ketamine (0.5 microM, 24h) produces loss of phenotype of fast-spiking interneurons via NADPH-oxidase (Science 2007). These data suggests that ketamine have multiple molecular targets in hypnotic, analgesic and amnestic actions. Propofol has been shown to enhance two types of GABAergic inhibition: a synaptic form (phasic inhibition) regulating neural excitability via the activation of postsynaptic GABAA receptors by intermittent GABA release from presynaptic terminals ; and a persistent tonic form (tonic inhibition) generated by continuous activation of extrasynaptic GABAA receptors by low concentrations of ambient GABA. However, the roles of tonic inhibition in hypnotic actions of isoflurane and sevoflurane are less clear. In this mini review, the relative contributions of extrasynaptic GABA receptors in behavioral actions of isoflurane and sevoflurane will be discussed. PMID- 21626858 TI - [Effect sites of anesthetics in the central nervous system network--looking into the mechanisms for natural sleep and anesthesia]. AB - We showed the effect sites of anesthetics in the central nervous system (CNS) network. The thalamus is a key factor for loss of consciousness during natural sleep and anesthesia. Although the linkages among neurons within the CNS network in natural sleep are complicated, but sophisticated, the sleep mechanism has been gradually unraveled. Anesthesia disrupts the link-ages between cortical and thalamic neurons and among the cortical neurons, and thus it loses the integration of information derived from the arousal and sleep nuclei. It has been considered that anesthesia does not share the common pathway as natural sleep at the level of unconsciousness, because anesthetics have multiple effect sites within CNS network and may induce disintegration among neurons. Recent literatures have shown that the effects of anesthetics are specific rather than global in the brain. It is interesting to note that thalamic injection of anti potassium channel materials restored consciousness during inhalation anesthesia, and that the sedative components of certain intravenous anesthesia may share the same pathway as natural sleep. To explore the sensitivity and susceptibility loci for anesthetics in the thalamocortical neurons as well as arousal and sleep nuclei within CNS network may be an important task for future study. PMID- 21626859 TI - [Mechanism of anesthesia: view from the EEG during anesthesia]. AB - It is widely known that electroencephalogram (EEG) shows dramatic changes with increase of the concentration of anesthetic. It is considered that volatile anesthetics (i. e. isoflurane, sevoflurane), barbiturates, propofol show anesthetic effect by potentiating GABAA receptor. Changing patterns of EEG by these anesthetics are quite similar. In light anesthetic level, high frequency with low amplitude waves are dominant. With increase of anesthetic concentration, waves in alpha range (8-13 Hz) become dominant. In deeper levels, powers in alpha range then become smaller and theta or delta powers become dominant. With further deeper levels, EEG waveform changes into specific pattern so-called "burst and suppression", and finally it becomes flat. The author considers that prominent alpha power indicates adequate anesthetic level. However this is not always the required condition for adequate anesthesia, because alpha power never becomes larger in some patients even when the anesthetic level was judged as adequate by concentration dependent changing patterns of EEG. As EEG changes in relation to the concentration of anesthetic, it seems to be correlated with the level of consciousness. But EEG patterns during anesthesia are mainly determined by the condition of thalamic neurons, and it would merely indicate the level of hypnosis indirectly. PMID- 21626860 TI - [Anesthetic mechanisms revealed by functional brain imaging]. AB - Recent advancement in functional brain imaging techniques has revealed much of the global effects of general anesthetics on the human brain. General anesthetics preferentially suppress specific brain areas including the parietal association cortex and the thalamus, part of which appears to mirror the default mode network. Low-level sensory areas are relatively preserved and remain activated even under deep sedation by anesthetics. Functional connectivity analysis by resting-state functional magnetic resonance imaging has shown that general anesthetics moderately suppress functional connectivity of the default mode network. Midazolam-induced loss of consciousness is associated with remarkable suppression of cortico-cortical propagation of evoked currents. Overall, those results prompt us to hypothesize that general anesthetics induce loss of consciousness by disrupting the integrative properties of the cerebral cortex. PMID- 21626861 TI - [Electrophysiological approach to mechanisms for actions of general anesthetics]. AB - Although general anesthetics were first used more than 160 years ago, their mechanisms have remained mysterious. During the past decade, significant progress in our understanding of general anesthetic action at the cellular and network system levels has been made. Our recent work demonstrates (a) that intravenous anesthetics, but not volatile agents, enhance the discharge of GABA from presynaptic terminals, (b) that intravenous anesthetics produce frequency dependent modification (FDM) of anesthesia, and (c) that FDM is responsible for the unsuccessful immobilization or hypnosis during intravenous anesthesia. In addition, we review the development of hypothesis for anesthetic action, non specific versus specific action, cutoff phenomenon in n-alcohols, and anesthesiological approach to consciousness. PMID- 21626862 TI - [Anesthetic mechanisms in the spinal cord]. AB - The essential elements of anesthesia are : hypnosis, amnesia, analgesia, immobility, and inhibition of untoward reflexes. The spinal cord is responsible for the latter three. Suppression of excitatory transmission and stimulation of inhibitory transmission are the anesthetic mechanisms in the spinal cord. Each anesthetic, however, has a unique effect on the transmission systems in the spinal cord. Some exclusively suppress excitatory transmission or stimulate inhibitory transmission, and others have a dual effect. The minimum alveolar/anesthetic concentration (MAC) is spinally mediated. Furthermore neurons in the ventral horn of spinal cord seem to be more depressed by anesthetics than neurons in the dorsal horn of the spinal cord. The ventral spinal cord also has relation to spinal cord ischemia. Investigation of the neuroprotective effect against spinal ischemia as well as the anesthetic effect in the ventral spinal cord is a very important subject of research. PMID- 21626863 TI - [Mechanism of general anesthesia and antinociception]. AB - Nitrous oxide exhibits antinociceptive effects by inhibition of transmission of nociceptive information from the primary afferent neuron to the spinal cord neuron through the activation of the descending inhibitory pathway. Furthermore, nitrous oxide might directly modulate the nociceptive transmission at the dorsal horn of the spinal cord. Analysis of knockout mice and pharmacological analysis suggest that the K -opioid receptor and the nociceptin receptor are involved in the antinociceptive effects of nitrous oxide. To elucidate the molecular mechanism in pharmacological effects of nitrous oxide, further investigation is necessary. PMID- 21626864 TI - [Influences of general anesthetics on the developing mammalian brain]. AB - Recently, almost all kinds of general anesthetics currently used in human clinical anesthesia, have been shown to exert neurodegenerative effects such as apoptosis of neuronal cells during the rapid synaptogenesis of immature mammalian brains, and later neurocognitive impairment. There are several drugs or strategies to reduce this phenomenon such as alpha(2) agonist, xenon, melatonin, lithium, hypothermia and erythropoietin, but their safety and efficacy should be investigated much further. Some human studies have shown that surgery under general anesthesia in infancy is one of the risk factors of the impairment of neurocognitive function, but others including Dutch twin study have shown that it is not. Larger-sized prospective randomized studies in human such as SAFEKIDS (http://www.iars.org/safekids/) to ascertain if current clinical practice of general anesthesia impairs neurocognitive development of human neonates and infants, are expected. They will also clarify what kind of anesthetics and anesthetic strategies may be the risk factors of neurocognitive impairment in human neonates and infants. PMID- 21626865 TI - [Intraoperative, salvaged, autologous blood transfusion in oncologic surgery]. AB - Intraoperative, salvaged, autologous blood transfusions carried out with autotransfusion devices are commonly used for cardiovascular surgery, and also enable the treatment of massive hemorrhage in orthopedic and gynecologic surgeries to prevent potential complications of homologous blood transfusions, such as transmission of infection, immune reactions, and blood type incompatibility. Transfusion of salvaged blood in oncologic surgery may cause hematogenous metastasis and dissemination of malignant tumor cells. However, some investigators have reported that blood irradiation or filtration using leukocyte reduction filters can prevent contamination by malignant tumor cells. Intraoperative autotransfusion with the combination of blood irradiation and leukocyte reduction filters could be therefore a promising technique for the treatment of profuse hemorrhage in oncologic surgery. PMID- 21626866 TI - [Decision to avoid general anesthesia on 3 cases with anterior mediastinal masses]. AB - We report 3 cases of anterior mediastinal masses in which we avoided providing general anesthesia for a biopsy and a central venous catheter placement. In all cases, chest X-rays on admission showed mediastinal mass ratio (MMR) greater than 44% and thoracic computed tomographic scans demonstrated cross sectional area (CSA) of the trachea 60% less than expected and the main stem bronchi narrowing. We made a decision not to provide general anesthesia, considering the risk of airway obstruction after induction of general anesthesia. In case 1, a 6-year-old boy, preoperative corticosteroid therapy relieved respiratory complaints without improvement of MMR and %CSA. On hospital day 3 the patient developed airway obstruction during induction of anesthesia and the surgery was postponed. After 3 days of additional chemotherapy MMR decreased to 34% and %CSA increased to 94%. On day 6 surgery under general anesthesia was performed safely. In case 2, a 15 year-old boy presented with MMR 44% and %CSA 48% and left bronchial stenosis and underwent surgery under local anesthesia. In case 3, a 3-year-old boy, preoperative corticosteroids and chemotherapy improved MMR 67% to 34% and %CSA 60% to 95%. On day 8 of admission a biopsy was performed under general anesthesia uneventfully. We emphasize not only clinical signs but also radiological signs are important to evaluate the safety in induction of general anesthesia for the management of the cases of anterior mediastinal masses. PMID- 21626867 TI - [Safety of preoperative oral rehydration therapy]. AB - BACKGROUND: OS-1 is an oral rehydration solution that conforms with the principles of oral rehydration therapy. It may be useful for preoperative fluid management of surgical patients. While intake of clear fluids 2 hours before surgery is considered safe, it is not known if the same applies to OS-1. We therefore investigated the safety of OS-1 for preoperative patients as compared with clear fluids. METHODS: First, eight healthy adult volunteers were studied in a crossover manner. Volunteers ingested 500 ml of OS-1 or water (clear fluid). Gastric emptying time was measured using gastric ultrasonography. Gastric antral area as measured by ultrasonography correlates well with gastric volume in a close-to-linear manner. Next, we measured gastric volume of elective surgical patients who had drunk OS-1 until two hours before the induction of anesthesia. RESULTS: Gastric emptying time did not differ between OS-1 and water. The stomach was emptied 30 minutes after ingestion of both OS-1 and water. The fasting stomach was identified in all patients who had drunk OS-1 before surgery. CONCLUSIONS: We concluded that allowing elective surgical patients to drink OS-1 until two hours before anesthesia did not affect the volume of gastric contents. PMID- 21626868 TI - [Case report: a normal dose of rocuronium achieved the desired effect in a short time after the administration of sugammadex during reoperation]. AB - A 69-year-old man with normal renal function underwent resection of a parotid tumor under general anesthesia. For tracheal intubation, rocuronium 0.6 mg x kg( 1) was administered, and for facial nerve stimulation, sugammadex 2 mg x kg(-1) was administered immediately after intubation. The operation time was 3 h. At the end of the surgery, sugammadex 2 mg x kg(-1) was administered again. Bleeding occurred 6 h after the surgery. During the second operation, rocuronium 0.6 mg x kg (-1) was administered for tracheal intubation. Maximal suppression was achieved 1 min 42 s after the administration of rocuronium, and the recovery time was 44 min. The times for both maximal suppression and recovery are similar to those when the same dose of rocuronium was used without sugammadex. The half-life of sugammadex is about 2 h. From the observations in this case, we think that after the completion of approximately 3 half-lives, a normal dose of rocuronium can produce the desired effect without the influence of residual sugammadex present in the plasma. PMID- 21626869 TI - [Atelectasis after induction of general anesthesia in an infant with cleft palate]. AB - We report an infant with cleft palate who developed atelectasis as a result of aspiration of food residue after induction of general anesthesia. A 12-month-old girl with cleft palate was scheduled for palatoplasty. The trachea was intubated after 6.5-hour fasting. Breath sounds were not audible on the left. Chest X-ray demonstrated complete atelectasis of the left lung. Bronchoscopy revealed that the left primary bronchus was blocked by a white stick. Pathological examination showed that the white stick consisted of milk residue with abundant candida species. It was speculated that the milk residue accumulated in the cleft palate had been aspirated during induction of general anesthesia. PMID- 21626870 TI - [Anesthetic management in a patient with trifunctional protein deficiency]. AB - A 4-year-old girl with trifunctional protein deficiency was scheduled for gastrostomy. She had recurrent episodes of rhabdomyolysis triggered by fasting, infection, stress and uncertain causes. In the management of anesthesia, we avoided both propofol and inhalational anesthetics and anesthetized her with benzodiazepine and opioid combined with regional anesthesia to minimize the stress response. Her metabolism was kept stable throughout the intraoperative period. However, on the second postoperative day, she developed rhabdomyolysis and was treated with glucose containing infusion. This case provides instructive information that strict management is needed for patients with trifunctional protein deficiency in the perioperative period. PMID- 21626871 TI - [Use of Parker Flex-Tip tracheal tube in a patient with Kniest dysplasia]. AB - A 37-year-old man with Kniest dysplasia and thoracic myelopathy was scheduled for T3-12 laminectomy. Kniest dysplasia is a severe chondrodysplasia characterized by short trunk and limbs. Awake fiberoptic intubation was attempted to prevent airway obstruction because we had predicted his difficult airway. During fiberoptic tracheal intubation, we easily succeeded in the insertion of the fiberscope itself into the trachea, but were not able to insert the tip of a reinforced tube into the trachea, because the tip of the reinforced tube impinged on laryngeal structures. We succeeded in placing the Parker Flex-Tip tracheal tube into the trachea. We considered that the Parker Flex-Tip tube, having a tip that reduces the gap between the fiberscope and the inside of the tube, resulted in success of the passage of the tube into the trachea during fiberoptic intubation. PMID- 21626872 TI - [Case of median nerve paralysis after hepatic segmentectomy]. AB - We report a case of a median nerve palsy. Hepatic segmentectomy and lymphnode dissection were performed in a 21-year-old man for multiple liver and retroperitoneal lymph nodes metastasis of seminoma. After surgery, patient complained of motor paralysis and hypesthesia of the left palm side of the thumb, first finger and radial half of the middle finger. He was diagnosed as having median nerve palsy. Motor paralysis and hypesthesia gradually disappeared over the two weeks after surgery. We should pay attention to appropriate positioning of the arm during surgery, and preoperative use of paclitaxel needs to be considered as etiology for perioperative peripheral nerve palsy. PMID- 21626873 TI - [Case of pulmonary hemorrhage suspected to be caused by pulmonary artery catheter after cardiac surgery]. AB - We report a case of pulmonary hemorrhage suspected to be caused by a pulmonary artery catheter (PAC) after cardiac surgery. An 80-year-old woman underwent aortic valve replacement, mitral valve replacement, and tricuspid annuloplasty. After anesthesia induction, a PAC was inserted from the right jugular vein and fixed at 48 cm, but it could not be wedged. Therefore, the PAC was fixed at 59 cm to measure the pulmonary capillary wedge pressure during the operation. The catheter showed a postoperative bending. The chest X-ray radiograph obtained at the first postoperative day showed a shadow in the right middle and lower lung. PAC was pulled and fixed at 48 cm, but the shadow continued to grow in size. A large volume of bloody drainage fluid was obtained. Therefore, we operated again. No bleeding or hematoma was observed in the mediastinum or pericardium. However, a hematoma spreading from the hilum of the right lung was observed under the visceral pleura. The patient was discharged on the 23rd postoperative day. The lung hemorrhage may have been caused by PAC-induced pulmonary artery injury since no lung injury was observed during reoperation. Therefore, care should be taken to prevent pulmonary artery injury by excessively deep insertion of PAC. PMID- 21626874 TI - [Validity of serum surfactant protein D measurements in a cross-sectional study]. AB - BACKGROUND: In October, 2008, the method for enzyme-linked immunoassay for serum surfactant protein D (SP-D) was changed from SP-D kit (Yamasa) EIA to SP-D kit (Yamasa) EIA2 in Japan. In our follow-up survey on 144 indium-exposed workers from December, 2008 to June, 2009, SP-D showed systematically higher values compared to the first survey from March to July, 2008. Geometric means (prevalence rate) of serum SP-D of the first and follow-up study were 52.1 ng/mL (4.1%) and 72.1 ng/mL (14.6%), respectively, though serum KL-6 and SP-A were stable. Thus, we examined the validity of the old and new methods. METHOD AND RESULTS: As a result of examining about 200 subjects, the correlation between the two methods was good. However, as a reproducibility examination of about another 300 subjects between the post-measured and the pre-measured values with the same new method at Laboratory A, the differences between vales in each observation are not acceptable. Especially, examinations performed using the lot 804, 805, and 105 at laboratory A showed systematic higher values than others. CONCLUSION: Various factors might have caused the large measurement bias; in particular, the new method lacks sufficient reproducibility between lots. When observing the chronological change of SP-D, attention must be given in comparisons of serum SP D after October, 2008 to before that because of the metrology-dependent measurement bias. PMID- 21626875 TI - [Clinical validity of renal function markers including serum cystatin C on chronic kidney disease classification]. AB - In this study, clinical utility of various renal function markers (Cystatin C, Creatinine, beta2-microglobulin, Urea nitrogen) including serum Cystatin C was evaluated by the latex turbidimetric immunoassay based on the Chronic Kidney Disease (CKD) stage classification. Serum creatinine was most correlated with estimated Glomerular Filtration Rate (eGFR), followed by serum Cystatin C. However, the level of serum Cystatin C increased significantly earlier than other renal function markers in the group of mild renal dysfunction based on the CKD stage classification. Cystatin C also had the most distinctive ability of the renal dysfunction by Receiver Operating Characteristic (ROC) and distinction characteristic analysis. Thus, it is useful to measure serum Cystatin C and serum creatinine at the same time to improve diagnostic sensitivity and specificity, eGFR was 6% higher than Ccr on stage III-V, though Ccr was 30% higher than eGFR on stage I-II by CKD stage classifications. Since eGFR and Ccr showed different results in early and severe CKD groups, it was suggested that Ccr and eGFR need to be evaluated as different renal function markers. PMID- 21626876 TI - [Usefulness of anti-SS-A/Ro antibody measurement based on fluorescence enzyme immunoassay with Ro60 and Ro52 antigen]. AB - OBJECTIVE: Anti-SS-A/Ro antibody is commonly found in the sera of patients with rheumatic disease. The antigenic components of SS-A/Ro are 60kD. and 52kD-Ro protein. Only anti-60kD SS-A/Ro antibody is detected by commonly used tests in clinical laboratories. "UniCAP EliA SS-A/Ro" is a new reagent using fluorescence enzyme immunoassay (FEIA) based on a mixture of both 60kD- and 52kD-Ro antigens. We evaluated the efficacy of detecting both anti-60kD and 52kD SS-A/Ro antibodies. PATIENTS AND METHODS: We collected sera from 264 rheumatic disease patients and 106 healthy subjects. Anti-SS-A/Ro antibodies were measured by the new reagent along with conventional method. Anti-52kD and 60kD SS-A/Ro antibodies were measured by ELISA kit in rheumatic disease patients. RESULTS: Anti-SS-A/Ro antibody levels were higher in patients with Sjogren's syndrome than those with SLE or RA. The prevalence of anti-SS-A/Ro antibody in rheumatic disease patients and healthy subjects were comparable with the conventional method, and patients with Sjogren's syndrome had highest prevalence of anti-SS-A/Ro antibody. Concordance rate between EliA and conventional method, and EliA and DID, were 96.6% and 94.3%, respectively. ELISA analysis revealed that patients with Sjogren's syndrome had anti-52kD SS-A/Ro antibody at high rates, while anti-60kD SS-A/Ro antibody was widely found in rheumatic disease patients. Three patients were positive only for anti-52kD SS-A/Ro antibody. CONCLUSION: Taken together, "UniCAP EliA SS-A/Ro" is useful as a screening test for anti-SS-A/Ro antibodies. PMID- 21626877 TI - [Development of a microbiology data warehouse (Akita-ReNICS) for networking hospitals in a medical region]. AB - The active involvement of hospital laboratory in surveillance is crucial to the success of nosocomial infection control. The recent dramatic increase of antimicrobial-resistant organisms and their spread into the community suggest that the infection control strategy of independent medical institutions is insufficient. To share the clinical data and surveillance in our local medical region, we developed a microbiology data warehouse for networking hospital laboratories in Akita prefecture. This system, named Akita-ReNICS, is an easy-to use information management system designed to compare, track, and report the occurrence of antimicrobial-resistant organisms. Participating laboratories routinely transfer their coded and formatted microbiology data to ReNICS server located at Akita University Hospital from their health care system's clinical computer applications over the internet. We established the system to automate the statistical processes, so that the participants can access the server to monitor graphical data in the manner they prefer, using their own computer's browser. Furthermore, our system also provides the documents server, microbiology and antimicrobiotic database, and space for long-term storage of microbiological samples. Akita-ReNICS could be a next generation network for quality improvement of infection control. PMID- 21626878 TI - [In-vitro diagnostics in Japan: its value and recognition--from the industry/providers' perspective]. AB - Since the 1960's in-vitro diagnostic tests (IVD) have been contributing greatly to the progress of medical care, which has lead to significant improvements in patients' quality of life. Today, it is apparent that IVD tests are an inevitable component of healthcare for individual treatment as well as national healthcare programs. Despite its significant contributions, the recognition for the value of IVD has been declining. This lack of recognition can be observed among healthcare professionals, governing bodies, as well as among patients, who are the immediate beneficiaries of innovative, and high quality IVD. As a result reimbursement levels for IVD testing have been reduced significantly by the National Health Institute (NHI) over the past 15 years. Recent activities to create a better understanding of the value of IVD have created less unfavorable NHI rate revisions in 2008 and 2010, but further strategic improvement plans will still need to be addressed with perseverance: 1) Enhancement of clinical value based NHI rate assignment processes, 2) Improvement of the accessibility to new/advanced IVD products, and 3) Public awareness programs to promote the value of IVD. There have been many efforts started to discuss critical issues of IVD within the Japanese healthcare system involving academics, IVD industry representation, and government officials (MHLW) since 2005. This article is describing and summarizing the identified and required priorities in order to create an environment that will allow to enhance Japanese healthcare through sustainable, reliable, and innovative IVD. PMID- 21626879 TI - [Ethical issues]. AB - Key concepts which should be recognized to understand today's medical ethics required for information management regarding clinical research are privacy protection, use limitation, individual participation, and accountability. Special attention should be paid to concepts other than privacy protection, because they are fairly new to medical professionals. Furthermore, in laboratory medicine, we have real problems, for example, how to protect privacy concerning specimens gathered from patients. Therefore, there have been many kinds of rules or guidelines established recently. Although we tend to strictly follow these guidelines, it is not always clear which guidelines should be applied to certain cases, or they do not always exactly correspond to a specific case. The full understanding of the principles of medical ethics represented by these guidelines is essential. In this paper, a clinical research document reviewed by an ethical review board is shown as an example. PMID- 21626880 TI - [Role of di-flavin reductase family in the regulation of nitric oxide synthase]. PMID- 21626881 TI - [Recent progress on the transporter studies by the constitutive biochemistry: a switch to regulate chemical transmission identified]. PMID- 21626882 TI - [Unique flavoenzyme: type 2 isopentenyl diphosphate isomerase]. PMID- 21626883 TI - [Negative regulation of RIG-I-like receptor (RLR) signaling]. PMID- 21626884 TI - [Role of arginine methylation in the Piwi-interacting RNA pathway]. PMID- 21626885 TI - [Sialic acid-deficient myopathy]. PMID- 21626886 TI - [New role of Cdk5 in oligodendrocyte development and dysmyelinating disorders of the central nervous system]. PMID- 21626887 TI - [Analysis of transporter function and development to clinical application]. PMID- 21626888 TI - [Self and non-self recognition through C-type lectin receptors]. PMID- 21626889 TI - [Application of multi-PK antibody that detects a wide variety of protein kinases]. PMID- 21626890 TI - [Rational combinations of antiepileptic drugs for refractory epilepsy]. AB - Although epilepsy surgery is most effective for patients with intractable epilepsy, a majority of them is not eligible for the surgery. Most of patients with refractory epilepsy are eventually treated with polypharmacy in hope of seizure control. Therefore, rational combinations of antiepileptic drugs are needed to control intractable seizures. Drug combinations should be rationally chosen based on the evidence of synergic efficacy and on avoidance of neurotoxicity. Several clinical studies suggest that the combination of valproate with lamotrigine has synergic antiepileptic effect. It has also been reported that the combination of carbamazepine with lamotrigine paradoxically decreases efficacy and increases toxicity. Animal studies using isobolography suggest that the combinations of topiramate with lamotrigine or levetiracetam are also promising on both seizure control and neurotoxicity. Clinical research is needed to examine these combinations. PMID- 21626891 TI - [Responsiveness glia-derived neurotrophic factor to nicotine in cultured rat cortical astrocytes]. PMID- 21626892 TI - Proceedings of the International Symposium on the Safe Use of Nanomaterials and Workshop on Nanomaterial Safety: Status, Procedures and Ethical Concerns. February 1-3, 2011. Lucknow, India. PMID- 21626893 TI - Should family physicians treat members of the same family?: YES. PMID- 21626894 TI - [The management table of the most recent SESP Congress]. PMID- 21626895 TI - A British perspective. PMID- 21626896 TI - Finding a better balance for the periodic health examination. PMID- 21626897 TI - Practical use of the Framingham risk score in primary prevention: Canadian perspective. AB - OBJECTIVE: To review the 2009 Canadian Cardiovascular Society guidelines and provide practical recommendations for physicians. SOURCES OF INFORMATION: Initial review of the references provided with the guidelines led to a search of the PubMed, ACP Journal Club, and Cochrane databases using the key words primary prevention and statin for English language clinical trials, randomized controlled trials, meta-analyses, and reviews conducted with human participants. References from appropriate retrieved articles were also reviewed. MAIN MESSAGE: The guidelines outline low-density lipoprotein cholesterol (LDL-C) thresholds and targets to inform optimal use of statins in the primary prevention of cardiovascular disease (CVD). Family history of CVD and levels of high sensitivity C-reactive protein (hsCRP) are risk modifiers in calculating the risk score with the new recommendations. An electronic calculator has been developed to facilitate increased uptake of these guidelines. Large numbers of asymptomatic people, particularly the elderly, will become eligible for statin therapy according to these new guidelines. Poor uptake by physicians and patients might result from the need for repeated testing of hsCRP and LDL-C levels in people who do not perceive themselves to be ill. Controversy persists concerning the role of hsCRP in the reclassification of CVD risk, and the concept of treating LDL-C to target has never been tested as an independent variable in a randomized trial. As two-thirds of the LDL-C lowering achieved by a statin occurs at the initial dose, it might be possible to achieve considerable CVD risk reduction for those at risk by treating initially with a mid-dose statin without LDL-C follow-up. CONCLUSION: A simplified approach might appeal to patients or physicians who find current guidelines too complex, cumbersome, or costly. Success in getting high-risk patients to take statins is key to achieving improved CVD mortality reduction. PMID- 21626898 TI - Muslim women having abortions in Canada: attitudes, beliefs, and experiences. AB - OBJECTIVE: To improve understanding of the attitudes, beliefs, and experiences of Muslim patients presenting for abortion. DESIGN: Exploratory study in which participants completed questionnaires about their attitudes, beliefs, and experiences. SETTING: Two urban, free-standing abortion clinics. PARTICIPANTS: Fifty-three self-identified Muslim patients presenting for abortion. MAIN OUTCOME MEASURES: Women's background, beliefs, and attitudes toward their religion and toward abortion; levels of anxiety, depression, and guilt, scored on a scale of 0 to 10; and degree of pro-choice or antichoice attitude toward abortion, assessed by having respondents identify under which circumstances a woman should be able to have an abortion. RESULTS: The 53 women in this study were a diverse group, aged 17 to 47 years, born in 17 different countries, with a range of beliefs and attitudes toward abortion. As found in previous studies, women who were less pro choice (identified fewer acceptable reasons to have an abortion) had higher anxiety and guilt scores than more pro-choice women did: 6.9 versus 4.9 (P = .01) and 6.9 versus 3.6 (P = .004), respectively. Women who said they strongly agreed that abortion was against Islamic principles also had higher anxiety and guilt scores: 9.3 versus 5.9 (P = .03) and 9.5 versus 5.3 (P = .03), respectively. CONCLUSION: Canadian Muslim women presenting for abortion come from many countries and schools of Islam. The group of Muslim women that we surveyed was so diverse that no generalizations can be made about them. Their attitudes toward abortion ranged from being completely prochoice to believing abortion is wrong unless it is done to save a woman's life. Many said they found their religion to be a source of comfort as well as a source of guilt, turning to prayer and meditation to cope with their feelings about the abortion. It is important that physicians caring for Muslim women understand that their patients come from a variety of backgrounds and can have widely differing beliefs. It might be helpful to be aware that patients who hold more anti-choice beliefs are likely to experience more anxiety and guilt related to their abortion than prochoice patients do. PMID- 21626899 TI - Family physician and obstetrician episiotomy rates in low-risk obstetrics in southern Alberta. AB - OBJECTIVE: To examine the episiotomy rate for women delivering in a regional hospital versus the rate in rural hospitals. DESIGN: Retrospective review of low risk delivery charts for a 12-month period (2006 to 2007). SETTING: One regional and 3 rural hospitals in southern Alberta. PARTICIPANTS: Charts were reviewed for a random sample of 10% of the women with low-risk deliveries at the regional hospital, and all such women at the participating rural hospitals. Eligible women were nulliparous or multiparous, were at 37 or more weeks' gestation, and delivered live newborns vaginally, including spontaneous and assisted vaginal deliveries. Low-risk deliveries were defined by the absence of high-risk maternal, prenatal, and perinatal features. MAIN OUTCOME MEASURES: Details of the delivery, including use of episiotomy. RESULTS: Charts were reviewed for 115 women who delivered in the regional hospital and for 140 women from the rural hospitals. Maternal and infant characteristics did not differ between settings (mean age 26 years, median parity 1, mean birth weight 3433 g [regional] and 3462 g [rural], and mean head circumference 35 cm). Episiotomies were performed in 13% of regional and 4% of rural deliveries (P = .01). Perineal tears occurred in 65% of regional (3 with third- to fourth-degree tears) and 57% of rural (2 with third to fourth-degree tears) deliveries (P = .20). Deliveries were carried out by 12 FPs and 6 obstetricians in the regional centre, and by 19 FPs in the rural hospitals. CONCLUSION: In our study, both rural and regional practitioners in southern Alberta demonstrated a "restrictive" use of episiotomy, in keeping with current evidence-based guidelines. Further prospective research is needed to examine how physician, maternal, and pregnancy characteristics affect episiotomy and perineal tear rates. PMID- 21626900 TI - Watching death. PMID- 21626901 TI - Doctors in Havana: witnessing their complex world. PMID- 21626902 TI - Teaching medical learners to appreciate "difficult" patients. PMID- 21626903 TI - Becoming what we need to be. PMID- 21626905 TI - [Health service research (7) Health service research and health care insurance service]. PMID- 21626904 TI - [Social factors determining health (10) Social capital]. PMID- 21626906 TI - [Public health for scientific study of society and health (3) "Communication through environmental epidemiology"]. PMID- 21626907 TI - Point/counterpoint. The terminal M.S. degree is no longer appropriate for students interested in a career in clinical medical physics in the United States. PMID- 21626908 TI - Internal margin assessment using cine MRI analysis of deglutition in head and neck cancer radiotherapy. AB - PURPOSE: Intensity-modulated radiation therapy (IMRT) is a promising treatment modality for patients with head and neck cancer (HNC). The dose distributions from IMRT are static and, thus, are unable to account for variations and/or uncertainties in the relationship between the patient (region being treated) and the beam. Organ motion comprises one such source of this uncertainty, introduced by physiological variation in the position, size, and shape of organs during treatment. In the head and neck, the predominant source of this variation arises from deglutition (swallowing). The purpose of this study was to investigate whether cinematographic MRI (cine MRI) could be used to determine asymmetric (nonuniform) internal margin (IM) components of tumor planning target volumes based on the actual deglutition-induced tumor displacement. METHODS: Five head and neck cancer patients were set up in treatment position on a 3 T MRI scanner. Two time series of single-slice, sagittal, cine images were acquired using a 2D FLASH sequence. The first time series was a 12.8 min scan designed to capture the frequency and duration of deglutition in the treatment position. The second time series was a short, 15 s scan designed to capture the displacement of deglutition in the treatment position. Deglutition frequency and mean swallow duration were estimated from the long time series acquisition. Swallowing and resting (nonswallowing) events were identified on the short time series acquisition and displacement was estimated based on contours of gross tumor volume (GTV) generated at each time point of a particular event. A simple linear relationship was derived to estimate 1D asymmetric IMs in the presence of resting- and deglutition-induced displacement. RESULTS: Deglutition was nonperiodic, with frequency and duration ranging from 2.89-24.18 mHz and from 3.86 to 6.10 s, respectively. The deglutition frequency and mean duration were found to vary among patients. Deglutition-induced maximal GTV displacements ranged from 0.00 to 28.36 mm with mean and standard deviation of 4.72 +/- 3.18, 3.70 +/- 2.81, 2.75 +/- 5.24, and 10.40 +/- 10.76 mm in the A, P, I, and S directions, respectively. Resting-induced maximal GTV displacement ranged from 0.00 to 5.59 mm with mean and standard deviation of 3.01 +/- 1.80, 1.25 +/- 1.10, 3.23 +/- 2.20, and 2.47 +/- 1.11 mm in the A, P, I, and S directions, respectively. For both resting and swallowing states, displacement along the S-I direction dominated displacement along the A-P direction. The calculated IMs were dependent on deglutition frequency, ranging from 3.28-4.37 mm for the lowest deglutition frequency patient to 3.76-6.43 mm for the highest deglutition frequency patient. A statistically significant difference was detected between IMs calculated for P and S directions (p = 0.0018). CONCLUSIONS: Cine MRI is able to capture tumor motion during deglutition. Swallowing events can be demarcated by MR signal intensity changes caused by anatomy containing fully relaxed spins that move medially into the imaging plane during deglutition. Deglutition is nonperiodic and results in dynamic changes in the tumor position. Deglutition-induced displacements are larger and more variable than resting displacements. The nonzero mean maximum resting displacement indicates that some tumor motion occurs even when the patient is not swallowing. Asymmetric IMs, derived from deglutition frequency, duration, and directional displacement, should be employed to account for tumor motion in HNC RT. PMID- 21626909 TI - A spatially encoded dose difference maximal intensity projection map for patient dose evaluation: a new first line patient quality assurance tool. AB - PURPOSE: To develop a spatially encoded dose difference maximal intensity projection (DD-MIP) as an online patient dose evaluation tool for visualizing the dose differences between the planning dose and dose on the treatment day. METHODS: Megavoltage cone-beam CT (MVCBCT) images acquired on the treatment day are used for generating the dose difference index. Each index is represented by different colors for underdose, acceptable, and overdose regions. A maximal intensity projection (MIP) algorithm is developed to compress all the information of an arbitrary 3D dose difference index into a 2D DD-MIP image. In such an algorithm, a distance transformation is generated based on the planning CT. Then, two new volumes representing the overdose and underdose regions of the dose difference index are encoded with the distance transformation map. The distance encoded indices of each volume are normalized using the skin distance obtained on the planning CT. After that, two MIPs are generated based on the underdose and overdose volumes with green-to-blue and green-to-red lookup tables, respectively. Finally, the two MIPs are merged with an appropriate transparency level and rendered in planning CT images. RESULTS: The spatially encoded DD-MIP was implemented in a dose-guided radiotherapy prototype and tested on 33 MVCBCT images from six patients. The user can easily establish the threshold for the overdose and underdose. A 3% difference between the treatment and planning dose was used as the threshold in the study; hence, the DD-MIP shows red or blue color for the dose difference > 3% or < or = 3%, respectively. With such a method, the overdose and underdose regions can be visualized and distinguished without being overshadowed by superficial dose differences. CONCLUSIONS: A DD-MIP algorithm was developed that compresses information from 3D into a single or two orthogonal projections while hinting the user whether the dose difference is on the skin surface or deeper. PMID- 21626910 TI - Analysis of Fourier-domain task-based detectability index in tomosynthesis and cone-beam CT in relation to human observer performance. AB - PURPOSE: Design and optimization of medical imaging systems benefit from accurate theoretical modeling that identifies the physical factors governing image quality, particularly in the early stages of system development. This work extends Fourier metrics of imaging performance and detectability index (d') to tomosynthesis and cone-beam CT (CBCT) and investigates the extent to which d' is a valid descriptor of task-based imaging performance as assessed by human observers, METHODS: The detectability index for tasks presented in 2D slices (d'(slice)) was derived from 3D cascaded systems analysis of tomosynthesis and CBCT. Anatomical background noise measured in a physical phantom presenting power law spectral density was incorporated in the "generalized" noise-equivalent quanta. Theoretical calculations of d'(slice) were performed as a function of total angular extent (theta(tot)) of source-detector orbit ranging 10 degrees - 360 degrees under two acquisition schemes: (i) Constant angular separation between projections (constant-delta theta), giving variable number of projections (N(proj)) and dose vs theta(tot) and (ii) constant number of projections (constant-N(proj)), giving constant dose (but variable angular sampling) with theta(tot). Five simple observer models were investigated: Prewhitening (PW), prewhitening with eye filter and internal noise (PWEi), nonprewhitening (NPW), nonprewhitening with eye filter (NPWE), and nonprewhitening with eye filter and internal noise (NPWEi). Human observer performance was measured in 9AFC tests for five simple imaging tasks presented within uniform and power-law clutter backgrounds. Measurements (from 9AFC tests) and theoretical calculations (from cascaded systems analysis of d'(slice)) were compared in terms of area under the ROC curve (A(z)) RESULTS: Reasonable correspondence between theoretical calculations and human observer performance was achieved for all imaging tasks over the broad range of experimental conditions and acquisition schemes. The PW and PWEi observer models tended to overestimate detectability, while the various NPW models predicted observer performance fairly well, with NPWEi giving the best overall agreement. Detectability was shown to increase with theta(tot) due to the reduction of out-of-plane clutter, reaching a plateau after a particular theta(tot) that depended on the imaging task. Depending on the acquisition scheme, however (i.e., constant-N(proj) or delta theta), detectability was seen in some cases to decline at higher theta(tot) due to tradeoffs among quantum noise, background clutter, and view sampling. CONCLUSIONS: Generalized detectability index derived from a 3D cascaded systems model shows reasonable correspondence with human observer performance over a fairly broad range of imaging tasks and conditions, although discrepancies were observed in cases relating to orbits intermediate to 180 degrees and 360 degrees. The basic correspondence of theoretical and measured performance supports the application of such a theoretical framework for system design and optimization of tomosynthesis and CBCT. PMID- 21626911 TI - Assisted hepatic resection using a toroidal HIFU device: an in vivo comparative study in pig. AB - PURPOSE: Bleeding is the main cause of postoperative complications during hepatic surgery. Blood loss and transfusions increase tumor recurrence in liver metastases from colorectal cancer. A high intensity focused ultrasound (HIFU) device with an integrated ultrasound imaging probe was developed for the treatment of colorectal liver metastasis. METHODS: The HIFU toroidal-shaped transducer contains 256 elements (working frequency: 3 MHz) and can create a single conical lesion of 7 cm3 in 40 s. Then, the volume of treatment can be significantly increased by juxtaposing single lesions. Presented here is the use of this device in an animal model as a complementary tool to improve surgical resection in the liver. Before transecting the liver, a wall of coagulative necrosis was performed using this device in order to minimize blood loss and dissection time during hepatectomy. Resection assisted by HIFU was compared to classical dissections with clamping [intermittent Pringle maneuver (IPM) group] and without clamping (control group). For each technique, 14 partial liver resections were performed in seven pigs. Blood loss per dissection surface area and resection time were the main outcome parameters. RESULTS: Conserving liver blood inflow during hepatic resection assisted by HIFU did not increase total blood loss (7.4 +/- 3.3 ml cm(-2)) compared to hepatic resection performed during IPM and controlled blood inflow (11.2 +/- 2.2 ml cm(-2)). Lower blood loss was measured on average when using HIFU, even though difference with clamping (IPM) was not statistically significant (p = 0.09). Resection assisted by HIFU reduced blood loss by 50% compared to control group (14.0 +/- 3.4 ml cm(-2), p = 0.03). The duration of transection when using HIFU (13 +/- 3 min) was significantly lower compared to clamping (23 +/- 4 min, p < 0.01) and control (18 +/- 3 min, p = 0.02). Precoagulation also resulted in sealing blood vessels with a diameter of less than 5 mm, and therefore the number of clips needed in the HIFU group was significantly lower (0.8 +/- 0.2 cm(-2)) when compared to clamping (1.6 +/- 0.2 cm(-2), p < 0.01) and control (1.8 +/- 0.4 cm(-2), p < 0.01). CONCLUSIONS: This method holds promise for future clinical applications in resection of liver metastases. PMID- 21626912 TI - An analysis of confidence limit calculations used in AAPM Task Group No. 119. AB - PURPOSE: The report issued by AAPM Task Group No. 119 outlined a procedure for evaluating the effectiveness of IMRT commissioning. The procedure involves measuring gamma pass-rate indices for IMRT plans of standard phantoms and determining if the results fall within a confidence limit set by assuming normally distributed data. As stated in the TG report, the assumption of normally distributed gamma pass rates is a convenient approximation for commissioning purposes, but may not accurately describe the data. Here the authors attempt to better describe gamma pass-rate data by fitting it to different distributions. The authors then calculate updated confidence limits using those distributions and compare them to those derived using TG No. 119 method. METHODS: Gamma pass rate data from 111 head and neck patients are fitted using the TG No. 119 normal distribution, a truncated normal distribution, and a Weibull distribution. Confidence limits to 95% are calculated for each and compared. A more general analysis of the expected differences between the TG No. 119 method of determining confidence limits and a more time-consuming curve fitting method is performed. RESULTS: The TG No. 119 standard normal distribution does not fit the measured data. However, due to the small range of measured data points, the inaccuracy of the fit has only a small effect on the final value of the confidence limits. The confidence limits for the 111 patient plans are within 0.1% of each other for all distributions. The maximum expected difference in confidence limits, calculated using TG No. 119's approximation and a truncated distribution, is 1.2%. CONCLUSIONS: A three-parameter Weibull probability distribution more accurately fits the clinical gamma index pass-rate data than the normal distribution adopted by TG No. 119. However, the sensitivity of the confidence limit on distribution fit is low outside of exceptional circumstances. PMID- 21626913 TI - Demons deformable registration of CT and cone-beam CT using an iterative intensity matching approach. AB - PURPOSE: A method of intensity-based deformable registration of CT and cone-beam CT (CBCT) images is described, in which intensity correction occurs simultaneously within the iterative registration process. The method preserves the speed and simplicity of the popular Demons algorithm while providing robustness and accuracy in the presence of large mismatch between CT and CBCT voxel values ("intensity"). METHODS: A variant of the Demons algorithm was developed in which an estimate of the relationship between CT and CBCT intensity values for specific materials in the image is computed at each iteration based on the set of currently overlapping voxels. This tissue-specific intensity correction is then used to estimate the registration output for that iteration and the process is repeated. The robustness of the method was tested in CBCT images of a cadaveric head exhibiting a broad range of simulated intensity variations associated with x-ray scatter, object truncation, and/or errors in the reconstruction algorithm. The accuracy of CT-CBCT registration was also measured in six real cases, exhibiting deformations ranging from simple to complex during surgery or radiotherapy guided by a CBCT-capable C-arm or linear accelerator, respectively. RESULTS: The iterative intensity matching approach was robust against all levels of intensity variation examined, including spatially varying errors in voxel value of a factor of 2 or more, as can be encountered in cases of high x-ray scatter. Registration accuracy without intensity matching degraded severely with increasing magnitude of intensity error and introduced image distortion. A single histogram match performed prior to registration alleviated some of these effects but was also prone to image distortion and was quantifiably less robust and accurate than the iterative approach. Within the six case registration accuracy study, iterative intensity matching Demons reduced mean TRE to (2.5 +/- 2.8) mm compared to (3.5 +/- 3.0) mm with rigid registration. CONCLUSIONS: A method was developed to iteratively correct CT-CBCT intensity disparity during Demons registration, enabling fast, intensity-based registration in CBCT-guided procedures such as surgery and radiotherapy, in which CBCT voxel values may be inaccurate. Accurate CT-CBCT registration in turn facilitates registration of multimodality preoperative image and planning data to intraoperative CBCT by way of the preoperative CT, thereby linking the intraoperative frame of reference to a wealth of preoperative information that could improve interventional guidance. PMID- 21626914 TI - Normal tissue dose conformality measures to guide radiotherapy fractionation decisions. AB - PURPOSE: To determine conditions under which hypofractionation could be favorable for a normal tissue--even if tumor [alpha/beta] exceeds the normal tissue's [alpha/beta]. METHODS: The hypofractionation sufficiency condition (HSC) for an organ is defined as a dose conformality constraint such that, if satisfied, a family of tumor control probability isoeffective fractionation schemes will show decreasing normal tissue complication probability with decreasing number of fractions. RESULTS: In the extended equivalent uniform dose (EUD) model [obtained by replacing dose with linear quadratic (LQ) 2 Gy equivalent dose], the HSC for a normal organ is proven to be satisfied if a suitably weighted average of the relative dose [hypofractionation sufficiency index (HSI)] is less than the ratio of normal tissue to tumor [alpha/beta]. The HSI is determined solely by dose distribution and the normal tissue volume factor, "a." If the HSC is satisfied for every normal tissue of concern, then there is a therapeutic gain with hypofractionation. The corresponding multifractionation sufficiency condition (therapeutic gain with increasing number of fractions) and multifractionation sufficiency index (MSI) are also derived. A sample clinical case is presented. CONCLUSIONS: Within the context of the LQ/EUD models, conformality measures (HSI and MSI) can be used to inform fractionation decisions. PMID- 21626915 TI - Optimization of a parallel hole collimator/CdZnTe gamma-camera architecture for scintimammography. AB - PURPOSE: Small field-of-view CdZnTe (CZT) gamma cameras are increasingly studied for breast lesion detection to complement mammography or ultrasonographic findings. However, in classical collimation configurations, they remain limited by the trade-off between spatial resolution and sensitivity. The HiSens architecture was proposed to overcome these limitations. Using an accurate 3D localization of the interactions inside the detector, this architecture leads to a gain in sensitivity without loss in spatial resolution. In this article, the relevance of the HiSens architecture for planar scintimammography is studied. METHODS: A detective quantum efficiency (DQE) computation method is developed and used to optimize the dimensioning of a parallel hole collimator dedicated to scintimammography. Based on the DQE curves, the impact of the collimator-to detector distance is studied. Two algorithms are proposed to combine data acquired with different collimator-to-detector distances. RESULTS: It is shown that CZT detector virtual pixelization increases system sensitivity by 3.3 while preserving a standard LEHR spatial resolution. The introduction of a gap between the CZT detector and the collimator is useful to modulate the DQE curve shape. The combination of data acquired using different gaps in the image formation process leads to enhanced restoration of the frequency content of the images, resulting in image contrast and spatial resolution improvements. CONCLUSIONS: Acquisition duration or injected activity could be markedly reduced if the HiSens architecture with an appropriate collimator-detector gap were used. PMID- 21626916 TI - Similarity evaluation in a content-based image retrieval (CBIR) CADx system for characterization of breast masses on ultrasound images. AB - PURPOSE: The authors are developing a content-based image retrieval (CBIR) CADx system to assist radiologists in characterization of breast masses on ultrasound images. In this study, the authors compared seven similarity measures to be considered for the CBIR system. The similarity between the query and the retrieved masses was evaluated based on radiologists' visual similarity assessments. METHODS: The CADx system retrieves masses that are similar to a query mass from a reference library based on computer-extracted features using a k-nearest neighbor (k-NN) approach. Among seven similarity measures evaluated for the CBIR system, four similarity measures including linear discriminant analysis (LDA), Bayesian neural network (BNN), cosine similarity measure (Cos), and Euclidean distance (ED) similarity measure were compared by radiologists' visual assessment. For LDA and BNN, the features of a query mass were combined first into a malignancy score and then masses with similar scores were retrieved. For Cos and ED, similar masses were retrieved based on the normalized dot product and the Euclidean distance, respectively, between two feature vectors. For the observer study, three most similar masses were retrieved for a given query mass with each method. All query-retrieved mass pairs were mixed and presented to the radiologists in random order. Three Mammography Quality Standards Act (MQSA) radiologists rated the similarity between each pair using a nine-point similarity scale (1 = very dissimilar, 9 = very similar). The accuracy of the CBIR CADx system using the different similarity measures to characterize malignant and benign masses was evaluated by ROC analysis. RESULTS: The BNN measure used with the k-NN classifier provided slightly higher performance for classification of malignant and benign masses (A(z) values of 0.87) than those with the LDA, Cos, and ED measures (A(z) of 0.86, 0.84, and 0.81, respectively). The average similarity ratings of all radiologists for LDA, BNN, Cos, and ED were 4.71, 4.95, 5.18, and 5.32, respectively. The k-NN with the ED measures retrieved masses of significantly higher similarity (p < 0.008) than LDA and BNN. CONCLUSIONS: Similarity measures using the resemblance of individual features in the multidimensional feature space can retrieve visually more similar masses than similarity measures using the resemblance of the classifier scores. A CBIR system that can most effectively retrieve similar masses to the query may not have the best A(z). PMID- 21626917 TI - Evaluation of intersession 3D-TRUS to 3D-TRUS image registration for repeat prostate biopsies. AB - PURPOSE: 3D-TRUS-guided prostate biopsy permits a 3D record of biopsy cores, supporting the planning of targets to resample or avoid during repeat biopsy sessions. Image registration is required in order to map biopsy targets planned on a previous session's 3D-TRUS image into the context of the current session. The authors evaluated the performance of surface- and intensity-based rigid and nonrigid registration algorithms for this task using a clinically motivated success criterion of a maximum 2.5 mm target registration error (TRE). METHODS: The authors collected two 3D-TRUS images for each of 13 patients, where each image was collected in a separate biopsy session, and the sessions were 1 week apart. The authors tested the iterative closest point and thin-plate spline surface-based registration methods, and the block matching and B-spline intensity based methods. Manually marked intrinsic fiducials (calcifications) were used to calculate a TRE for each of the tested methods. In addition, error ellipsoids, anisotropy, and variability due to image segmentation were analyzed. All analysis was performed separately for the peripheral zone since this area harbors up to 80% of all prostate cancer. RESULTS: Only the intensity-based nonrigid registration method met the success criterion for both the whole gland and the peripheral zone. Segmentation was a substantial contributor to registration error variability for the surface-based methods, and the surface-based methods resulted in greater error volumes and anisotropy. CONCLUSIONS: Intensity-based rigid registration is clinically sufficient to register regions outside the peripheral zone, but nonrigid registration is required in order to register the peripheral zone with clinically needed accuracy. The clinical advantage of using nonrigid registration is questionable since the difference between the RMS TREs for rigid and nonrigid intensity-based registration could be considered to be small (0.3 mm) and is statistically significant. If the added clinical value in performing a nonrigid registration is insufficient given the additional time required for this computation, rigid registration alone may be suitable. PMID- 21626918 TI - Development and evaluation of a computer-aided diagnostic scheme for lung nodule detection in chest radiographs by means of two-stage nodule enhancement with support vector classification. AB - PURPOSE: To develop a computer-aided detection (CADe) scheme for nodules in chest radiographs (CXRs) with a high sensitivity and a low false-positive (FP) rate. METHODS: The authors developed a CADe scheme consisting of five major steps, which were developed for improving the overall performance of CADe schemes. First, to segment the lung fields accurately, the authors developed a multisegment active shape model. Then, a two-stage nodule-enhancement technique was developed for improving the conspicuity of nodules. Initial nodule candidates were detected and segmented by using the clustering watershed algorithm. Thirty one shape-, gray-level-, surface-, and gradient-based features were extracted from each segmented candidate for determining the feature space, including one of the new features based on the Canny edge detector to eliminate a major FP source caused by rib crossings. Finally, a nonlinear support vector machine (SVM) with a Gaussian kernel was employed for classification of the nodule candidates. RESULTS: To evaluate and compare the scheme to other published CADe schemes, the authors used a publicly available database containing 140 nodules in 140 CXRs and 93 normal CXRs. The CADe scheme based on the SVM classifier achieved sensitivities of 78.6% (110/140) and 71.4% (100/140) with averages of 5.0 (1165/233) FPs/image and 2.0 (466/233) FPs/image, respectively, in a leave-one out cross-validation test, whereas the CADe scheme based on a linear discriminant analysis classifier had a sensitivity of 60.7% (85/140) at an FP rate of 5.0 FPs/image. For nodules classified as "very subtle" and "extremely subtle," a sensitivity of 57.1% (24/42) was achieved at an FP rate of 5.0 FPs/image. When the authors used a database developed at the University of Chicago, the sensitivities was 83.3% (40/48) and 77.1% (37/48) at an FP rate of 5.0 (240/48) FPs/image and 2.0 (96/48) FPs/image, respectively. CONCLUSIONS: These results compare favorably to those described for other commercial and non-commercial CADe nodule detection systems. PMID- 21626919 TI - Dose response of alanine detectors irradiated with carbon ion beams. AB - PURPOSE: The dose response of the alanine detector shows a dependence on particle energy and type when irradiated with ion beams. The purpose of this study is to investigate the response behavior of the alanine detector in clinical carbon ion beams and compare the results to model predictions. METHODS: Alanine detectors have been irradiated with carbon ions with an energy range of 89-400 MeV/u. The relative effectiveness of alanine has been measured in this regime. Pristine and spread out Bragg peak depth-dose curves have been measured with alanine dosimeters. The track structure based alanine response model developed by Hansen and Olsen has been implemented in the Monte Carlo code FLUKA and calculations were compared to experimental results. RESULTS: Calculations of the relative effectiveness deviate less than 5% from the measured values for monoenergetic beams. Measured depth-dose curves deviate from predictions in the peak region, most pronounced at the distal edge of the peak. CONCLUSIONS: The used model and its implementation show a good overall agreement for quasi-monoenergetic measurements. Deviations in depth-dose measurements are mainly attributed to uncertainties of the detector geometry implemented in the Monte Carlo simulations. PMID- 21626920 TI - Computer-aided detection of breast masses: four-view strategy for screening mammography. AB - PURPOSE: To improve the performance of a computer-aided detection (CAD) system for mass detection by using four-view information in screening mammography. METHODS: The authors developed a four-view CAD system that emulates radiologists' reading by using the craniocaudal and mediolateral oblique views of the ipsilateral breast to reduce false positives (FPs) and the corresponding views of the contralateral breast to detect asymmetry. The CAD system consists of four major components: (1) Initial detection of breast masses on individual views, (2) information fusion of the ipsilateral views of the breast (referred to as two view analysis), (3) information fusion of the corresponding views of the contralateral breast (referred to as bilateral analysis), and (4) fusion of the four-view information with a decision tree. The authors collected two data sets for training and testing of the CAD system: A mass set containing 389 patients with 389 biopsy-proven masses and a normal set containing 200 normal subjects. All cases had four-view mammograms. The true locations of the masses on the mammograms were identified by an experienced MQSA radiologist. The authors randomly divided the mass set into two independent sets for cross validation training and testing. The overall test performance was assessed by averaging the free response receiver operating characteristic (FROC) curves of the two test subsets. The FP rates during the FROC analysis were estimated by using the normal set only. The jackknife free-response ROC (JAFROC) method was used to estimate the statistical significance of the difference between the test FROC curves obtained with the single-view and the four-view CAD systems. RESULTS: Using the single-view CAD system, the breast-based test sensitivities were 58% and 77% at the FP rates of 0.5 and 1.0 per image, respectively. With the four-view CAD system, the breast-based test sensitivities were improved to 76% and 87% at the corresponding FP rates, respectively. The improvement was found to be statistically significant (p < 0.0001) by JAFROC analysis. CONCLUSIONS: The four view information fusion approach that emulates radiologists' reading strategy significantly improves the performance of breast mass detection of the CAD system in comparison with the single-view approach. PMID- 21626921 TI - An alternating focused ultrasound system for thermal therapy studies in small animals. AB - PURPOSE: To develop an alternating focused ultrasound system (AFUS) for preclinical studies of thermal and acoustic responses of tumors in small animal models. This work was motivated by the need of noninvasively creating relatively small spheroidal thermal lesions in small targets (e.g., a murine tumor) without damaging the surrounding tissues. METHODS: The AFUS consists of two lead zirconate titanate (PZT-4) spherically curved ultrasound transducers with focal zones crossing each other at a 90 degrees angle. The transducers were independently powered following a programed alternating firing scheme. Before the device design and construction, an acoustic and biothermal model was developed to simulate the ultrasound pressure field and the resulting temperature and thermal dose distributions. A shape factor, sphericity, to quantify the roundness of the lesions was calculated based on the 240 equivalent minutes at 43 degrees C thermal dose contours. A prototype of the AFUS was constructed with two identical transducers of an operating frequency of 2.25 MHz, 38 mm in diameter, and F number equal to 1.33. To evaluate the performance of the AFUS experimentally, a series of heating in polyacrylamide phantoms, ex vivo porcine liver tissues, and in implanted mouse tumors fibrosarcoma (FSaII) in vivo was conducted. In these experimental cases, the sphericity was calculated and compared based on the visible lesion (a marked change in coloration). RESULTS: As shown in the simulations, the lesions induced in polyacrylamide phantoms, ex vivo porcine liver tissues, and in vivo mouse tumors, the sphericities of the lesions yielded by AFUS heating were approximately 50% higher than those of single focused ultrasound heating as long as moderate intensities were used and the duty cycle pulses were distributed equally among the transducers. CONCLUSIONS: The AFUS is a device capable of noninvasively creating spheroidal thermal lesions in small targets such as murine tumors. PMID- 21626922 TI - Massive-training support vector regression and Gaussian process for false positive reduction in computer-aided detection of polyps in CT colonography. AB - PURPOSE: A massive-training artificial neural network (MTANN) has been developed for the reduction of false positives (FPs) in computer-aided detection (CADe) of polyps in CT colonography (CTC). A major limitation of the MTANN is the long training time. To address this issue, the authors investigated the feasibility of two state-of-the-art regression models, namely, support vector regression (SVR) and Gaussian process regression (GPR) models, in the massive-training framework and developed massive-training SVR (MTSVR) and massive-training GPR (MTGPR) for the reduction of FPs in CADe of polyps. METHODS: The authors applied SVR and GPR as volume-processing techniques in the distinction of polyps from FP detections in a CTC CADe scheme. Unlike artificial neural networks (ANNs), both SVR and GPR are memory-based methods that store a part of or the entire training data for testing. Therefore, their training is generally fast and they are able to improve the efficiency of the massive-training methodology. Rooted in a maximum margin property, SVR offers excellent generalization ability and robustness to outliers. On the other hand, GPR approaches nonlinear regression from a Bayesian perspective, which produces both the optimal estimated function and the covariance associated with the estimation. Therefore, both SVR and GPR, as the state-of-the-art nonlinear regression models, are able to offer a performance comparable or potentially superior to that of ANN, with highly efficient training. Both MTSVR and MTGPR were trained directly with voxel values from CTC images. A 3D scoring method based on a 3D Gaussian weighting function was applied to the outputs of MTSVR and MTGPR for distinction between polyps and nonpolyps. To test the performance of the proposed models, the authors compared them to the original MTANN in the distinction between actual polyps and various types of FPs in terms of training time reduction and FP reduction performance. The authors' CTC database consisted of 240 CTC data sets obtained from 120 patients in the supine and prone positions. The training set consisted of 27 patients, 10 of which had 10 polyps. The authors selected 10 nonpolyps (i.e., FP sources) from the training set. These ten polyps and ten nonpolyps were used for training the proposed models. The testing set consisted of 93 patients, including 19 polyps in 7 patients and 86 negative patients with 474 FPs produced by an original CADe scheme. RESULTS: With the MTSVR, the training time was reduced by a factor of 190, while a FP reduction performance [by-polyp sensitivity of 94.7% (18/19) with 2.5 (230/93) FPs/patient] comparable to that of the original MTANN [the same sensitivity with 2.6 (244/93) FPs/patient] was achieved. The classification performance in terms of the area under the receiver-operating-characteristic curve value of the MTGPR (0.82) was statistically significantly higher than that of the original MTANN (0.77), with a two-sided p-value of 0.03. The MTGPR yielded a 94.7% (18/19) by-polyp sensitivity at a FP rate of 2.5 (235/93) per patient and reduced the training time by a factor of 1.3. CONCLUSIONS: Both MTSVR and MTGPR improve the efficiency of the training in the massive-training framework while maintaining a comparable performance. PMID- 21626923 TI - Proton radiography and fluoroscopy of lung tumors: a Monte Carlo study using patient-specific 4DCT phantoms. AB - PURPOSE: Monte Carlo methods are used to simulate and optimize a time-resolved proton range telescope (TRRT) in localization of intrafractional and interfractional motions of lung tumor and in quantification of proton range variations. METHODS: The Monte Carlo N-Particle eXtended (MCNPX) code with a particle tracking feature was employed to evaluate the TRRT performance, especially in visualizing and quantifying proton range variations during respiration. Protons of 230 MeV were tracked one by one as they pass through position detectors, patient 4DCT phantom, and finally scintillator detectors that measured residual ranges. The energy response of the scintillator telescope was investigated. Mass density and elemental composition of tissues were defined for 4DCT data. RESULTS: Proton water equivalent length (WEL) was deduced by a reconstruction algorithm that incorporates linear proton track and lateral spatial discrimination to improve the image quality. 4DCT data for three patients were used to visualize and measure tumor motion and WEL variations. The tumor trajectories extracted from the WEL map were found to be within 1 mm agreement with direct 4DCT measurement. Quantitative WEL variation studies showed that the proton radiograph is a good representation of WEL changes from entrance to distal of the target. CONCLUSIONS: MCNPX simulation results showed that TRRT can accurately track the motion of the tumor and detect the WEL variations. Image quality was optimized by choosing proton energy, testing parameters of image reconstruction algorithm, and comparing to ground truth 4DCT. The future study will demonstrate the feasibility of using the time resolved proton radiography as an imaging tool for proton treatments of lung tumors. PMID- 21626924 TI - Dynamic gating window for compensation of baseline shift in respiratory-gated radiation therapy. AB - PURPOSE: To analyze and evaluate the necessity and use of dynamic gating techniques for compensation of baseline shift during respiratory-gated radiation therapy of lung tumors. METHODS: Motion tracking data from 30 lung tumors over 592 treatment fractions were analyzed for baseline shift. The finite state model (FSM) was used to identify the end-of-exhale (EOE) breathing phase throughout each treatment fraction. Using duty cycle as an evaluation metric, several methods of end-of-exhale dynamic gating were compared: An a posteriori ideal gating window, a predictive trend-line-based gating window, and a predictive weighted point-based gating window. These methods were evaluated for each of several gating window types: Superior/inferior (SI) gating, anterior/posterior beam, lateral beam, and 3D gating. RESULTS: In the absence of dynamic gating techniques, SI gating gave a 39.6% duty cycle. The ideal SI gating window yielded a 41.5% duty cycle. The weight-based method of dynamic SI gating yielded a duty cycle of 36.2%. The trend-line-based method yielded a duty cycle of 34.0%. CONCLUSIONS: Dynamic gating was not broadly beneficial due to a breakdown of the FSM's ability to identify the EOE phase. When the EOE phase was well defined, dynamic gating showed an improvement over static-window gating. PMID- 21626925 TI - Calibration of GafChromic XR-RV3 radiochromic film for skin dose measurement using standardized x-ray spectra and a commercial flatbed scanner. AB - PURPOSE: In this study, newly formulated XR-RV3 GafChromic film was calibrated with National Institute of Standards and Technology (NIST) traceability for measurement of patient skin dose during fluoroscopically guided interventional procedures. METHODS: The film was calibrated free-in-air to air kerma levels between 15 and 1100 cGy using four moderately filtered x-ray beam qualities (60, 80, 100, and 120 kVp). The calibration films were scanned with a commercial flatbed document scanner. Film reflective density-to-air kerma calibration curves were constructed for each beam quality, with both the orange and white sides facing the x-ray source. A method to correct for nonuniformity in scanner response (up to 25% depending on position) was developed to enable dose measurement with large films. The response of XR-RV3 film under patient backscattering conditions was examined using on-phantom film exposures and Monte Carlo simulations. RESULTS: The response of XR-RV3 film to a given air kerma depended on kVp and film orientation. For a 200 cGy air kerma exposure with the orange side of the film facing the source, the film response increased by 20% from 60 to 120 kVp. At 500 cGy, the increase was 12%. When 500 cGy exposures were performed with the white side facing the x-ray source, the film response increased by 4.0% (60 kVp) to 9.9% (120 kVp) compared to the orange-facing orientation. On-phantom film measurements and Monte Carlo simulations show that using a NIST-traceable free-in-air calibration curve to determine air kerma in the presence of backscatter results in an error from 2% up to 8% depending on beam quality. The combined uncertainty in the air kerma measurement from the calibration curves and scanner nonuniformity correction was +/- 7.1% (95% C.I.). The film showed notable stability. Calibrations of film and scanner separated by 1 yr differed by 1.0%. CONCLUSIONS: XR-RV3 radiochromic film response to a given air kerma shows dependence on beam quality and film orientation. The presence of backscatter slightly modifies the x-ray energy spectrum; however, the increase in film response can be attributed primarily to the increase in total photon fluence at the sensitive layer. Film calibration curves created under free-in-air conditions may be used to measure dose from fluoroscopic quality x-ray beams, including patient backscatter with an error less than the uncertainty of the calibration in most cases. PMID- 21626926 TI - Multisource modeling of flattening filter free (FFF) beam and the optimization of model parameters. AB - PURPOSE: With the introduction of flattening filter free (FFF) linear accelerators to radiation oncology, new analytical source models for a FFF beam applicable to current treatment planning systems is needed. In this work, a multisource model for the FFF beam and the optimization of involved model parameters were designed. METHODS: The model is based on a previous three source model proposed by Yang et al. ["A three-source model for the calculation of head scatter factors," Med. Phys. 29, 2024-2033 (2002)]. An off axis ratio (OAR) of photon fluence was introduced to the primary source term to generate cone shaped profiles. The parameters of the source model were determined from measured head scatter factors using a line search optimization technique. The OAR of the photon fluence was determined from a measured dose profile of a 40 x 40 cm2 field size with the same optimization technique, but a new method to acquire gradient terms for OARs was developed to enhance the speed of the optimization process. The improved model was validated with measured dose profiles from 3 x 3 to 40 x 40 cm2 field sizes at 6 and 10 MV from a TrueBeam STx linear accelerator. Furthermore, planar dose distributions for clinically used radiation fields were also calculated and compared to measurements using a 2D array detector using the gamma index method. RESULTS: All dose values for the calculated profiles agreed with the measured dose profiles within 0.5% at 6 and 10 MV beams, except for some low dose regions for larger field sizes. A slight overestimation was seen in the lower penumbra region near the field edge for the large field sizes by 1%-4%. The planar dose calculations showed comparable passing rates (> 98%) when the criterion of the gamma index method was selected to be 3%/3 mm. CONCLUSIONS: The developed source model showed good agreements between measured and calculated dose distributions. The model is easily applicable to any other linear accelerator using FFF beams as the required data include only the measured PDD, dose profiles, and output factors for various field sizes, which are easily acquired during conventional beam commissioning process. PMID- 21626927 TI - On the potential of ultrasound elastography for pressure ulcer early detection. AB - PURPOSE: Pressure ulcers are areas of soft tissue breakdown induced by a sustained mechanical stress that damages the skin and underlying tissues. They represent a considerable burden to the society in terms of health care and cost. Yet, techniques for prevention and detection of pressure ulcers still remain very limited. In this article, the authors investigated the potential of ultrasound elastography for pressure ulcer early detection. Elastography is an imaging technique providing local information on biological tissue mechanical properties. It is relevant for pressure ulcer detection as this pathology is associated with a gradual stiffening of damaged tissues, beginning in the deeper tissues and progressing toward the skin surface. METHODS: A 2D ultrasound elastography method was proposed and its ability in terms of pressure ulcer detection was validated through numerical simulations and physical acquisitions on pressure ulcer mimicking phantoms. In vivo experiments on a rat model are also reported. A maintained pressure was applied on the animal thigh, with a view to generate a pressure ulcer, and ultrasound data were acquired and processed before and after application of this pressure. RESULTS: Numerical simulations demonstrated that a pressure ulcer can theoretically be detected at a very early stage with ultrasound elastography. Even when the ulcer region was characterized by a low stiffening (ratio of 1.8 relative to normal tissues), the corresponding elastogram clearly underlined the pathological area. This observation was confirmed by the results obtained on a physical phantom mimicking a pressure ulcer at an early stage. Computed elastograms showed strain differences between areas mimicking healthy and pathological tissues. Results corresponding to in vivo experiments revealed a difference in the way tissues behaved before and after the pressure was applied on the animal thigh, which strongly suggests the presence of a pathological area. CONCLUSIONS: Experiments demonstrated that ultrasound elastography is a promising technique for pressure ulcer detection, especially at an early stage of the pathology, when the disease is still visually undetectable. In the absence of any gold standard method, this is also a first step toward the development of a quantitative technique. PMID- 21626928 TI - Deconvolution by finite-size-source effects of x-ray phase-contrast images. AB - PURPOSE: In the hard x-ray region, the cross sections for the phase shift of low Z elements are about 1000 times larger than the absorption ones. As a consequence, phase contrast is detectable even when absorption contrast is minimal or absent. Therefore, phase-contrast imaging could become a valid alternative to absorption contrast without delivering high dose to tissue/human body parts. METHODS: To enhance the quality of phase-contrast images without increasing the dose, a possible approach could be the partial deconvolution of the finite source size effects by experimental phase-contrast images. The deconvolution procedure, the authors propose, employs the acquisition of two images on a suitable well-known test sample, one in contact and the other in phase-contrast conditions. Both acquired images are used along with a simulated phase-contrast image (obtained from the test sample in ideal conditions of pointlike source illumination) to correctly retrieve the experimental source distribution function. This information allows a generic experimental phase contrast image, acquired in the same conditions, to be partially deconvolved by finite source size effects. RESULTS: The performed experimental tests indicate that deconvolved images are equivalent to those which would be obtained with a source 40% smaller than the actual size. In turn, this finding is equivalent to an increase of the "effective" lateral spatial coherence length. The corresponding quality improvement of the phase-contrast imaging is directly deducible by the presence of many Fresnel fringes, much better visible with respect to the original experimental phase-contrast images. CONCLUSIONS: The use of a test standard sample, always possible in every experimental setup, to partially deconvolve the finite-size-source blurring effects shows that higher quality phase-contrast images could be readily available, making easier diagnoses and tissue/sample analyses. The method could give, in the future, the possibility to further lower the delivered dose to patients, organs, and tissues when compact room-sized and brilliant microfocus x-ray sources will be available for clinical applications in hospitals. PMID- 21626929 TI - Computer-aided detection of lung nodules via 3D fast radial transform, scale space representation, and Zernike MIP classification. AB - PURPOSE: The authors presented a novel system for automated nodule detection in lung CT exams. METHODS: The approach is based on (1) a lung tissue segmentation preprocessing step, composed of histogram thresholding, seeded region growing, and mathematical morphology; (2) a filtering step, whose aim is the preliminary detection of candidate nodules (via 3D fast radial filtering) and estimation of their geometrical features (via scale space analysis); and (3) a false positive reduction (FPR) step, comprising a heuristic FPR, which applies thresholds based on geometrical features, and a supervised FPR, which is based on support vector machines classification, which in turn, is enhanced by a feature extraction algorithm based on maximum intensity projection processing and Zernike moments. RESULTS: The system was validated on 154 chest axial CT exams provided by the lung image database consortium public database. The authors obtained correct detection of 71% of nodules marked by all radiologists, with a false positive rate of 6.5 false positives per patient (FP/patient). A higher specificity of 2.5 FP/patient was reached with a sensitivity of 60%. An independent test on the ANODE09 competition database obtained an overall score of 0.310. CONCLUSIONS: The system shows a novel approach to the problem of lung nodule detection in CT scans: It relies on filtering techniques, image transforms, and descriptors rather than region growing and nodule segmentation, and the results are comparable to those of other recent systems in literature and show little dependency on the different types of nodules, which is a good sign of robustness. PMID- 21626930 TI - Comparative performance of multiview stereoscopic and mammographic display modalities for breast lesion detection. AB - PURPOSE: Mammography is known to be one of the most difficult radiographic exams to interpret. Mammography has important limitations, including the superposition of normal tissue that can obscure a mass, chance alignment of normal tissue to mimic a true lesion and the inability to derive volumetric information. It has been shown that stereomammography can overcome these deficiencies by showing that layers of normal tissue lay at different depths. If standard stereomammography (i.e., a single stereoscopic pair consisting of two projection images) can significantly improve lesion detection, how will multiview stereoscopy (MVS), where many projection images are used, compare to mammography? The aim of this study was to assess the relative performance of MVS compared to mammography for breast mass detection. METHODS: The MVS image sets consisted of the 25 raw projection images acquired over an arc of approximately 45 degrees using a Siemens prototype breast tomosynthesis system. The mammograms were acquired using a commercial Siemens FFDM system. The raw data were taken from both of these systems for 27 cases and realistic simulated mass lesions were added to duplicates of the 27 images at the same local contrast. The images with lesions (27 mammography and 27 MVS) and the images without lesions (27 mammography and 27 MVS) were then postprocessed to provide comparable and representative image appearance across the two modalities. All 108 image sets were shown to five full time breast imaging radiologists in random order on a state-of-the-art stereoscopic display. The observers were asked to give a confidence rating for each image (0 for lesion definitely not present, 100 for lesion definitely present). The ratings were then compiled and processed using ROC and variance analysis. RESULTS: The mean AUC for the five observers was 0.614 +/- 0.055 for mammography and 0.778 +/- 0.052 for multiview stereoscopy. The difference of 0.164 +/- 0.065 was statistically significant with a p-value of 0.0148. CONCLUSIONS: The differences in the AUCs and the p-value suggest that multiview stereoscopy has a statistically significant advantage over mammography in the detection of simulated breast masses. This highlights the dominance of anatomical noise compared to quantum noise for breast mass detection. It also shows that significant lesion detection can be achieved with MVS without any of the artifacts associated with tomosynthesis. PMID- 21626931 TI - Dosimetric accuracy of a deterministic radiation transport based 192Ir brachytherapy treatment planning system. Part II: Monte Carlo and experimental verification of a multiple source dwell position plan employing a shielded applicator. AB - PURPOSE: The aim of this work is the dosimetric validation of a deterministic radiation transport based treatment planning system (BRACHYVISION v. 8.8, referred to as TPS in the following) for multiple 192Ir source dwell position brachytherapy applications employing a shielded applicator in homogeneous water geometries. METHODS: TPS calculations for an irradiation plan employing seven VS2000 192Ir high dose rate (HDR) source dwell positions and a partially shielded applicator (GM11004380) were compared to corresponding Monte Carlo (MC) simulation results, as well as experimental results obtained using the VIP polymer gel-magnetic resonance imaging three-dimensional dosimetry method with a custom made phantom. RESULTS: TPS and MC dose distributions were found in agreement which is mainly within +/- 2%. Considerable differences between TPS and MC results (greater than 2%) were observed at points in the penumbra of the shields (i.e., close to the edges of the "shielded" segment of the geometries). These differences were experimentally verified and therefore attributed to the TPS. Apart from these regions, experimental and TPS dose distributions were found in agreement within 2 mm distance to agreement and 5% dose difference criteria. As shown in this work, these results mark a significant improvement relative to dosimetry algorithms that disregard the presence of the shielded applicator since the use of the latter leads to dosimetry errors on the order of 20%-30% at the edge of the "unshielded" segment of the geometry and even 2%-6% at points corresponding to the potential location of the target volume in clinical applications using the applicator (points in the unshielded segment at short distances from the applicator). CONCLUSIONS: Results of this work attest the capability of the TPS to accurately account for the scatter conditions and the increased attenuation involved in HDR brachytherapy applications employing multiple source dwell positions and partially shielded applicators. PMID- 21626932 TI - Subzone based magnetic resonance elastography using a Rayleigh damped material model. AB - PURPOSE: Recently, the attenuating behavior of soft tissue has been addressed in magnetic resonance elastography by the inclusion of a damping mechanism in the methods used to reconstruct the resulting mechanical property image. To date, this mechanism has been based on a viscoelastic model for material behavior. Rayleigh, or proportional, damping provides a more generalized model for elastic energy attenuation that uses two parameters to characterize contributions proportional to elastic and inertial forces. In the case of time-harmonic vibration, these two parameters lead to both the elastic modulus and the density being complex valued (as opposed to the case of pure viscoelasticity, where only the elastic modulus is complex valued). METHODS: This article presents a description of Rayleigh damping in the time-harmonic case, discussing the differences between this model and the viscoelastic damping models. In addition, the results from a subzone based Rayleigh damped elastography study of gelatin and tofu phantoms are discussed, along with preliminary results from in vivo breast data. RESULTS: Both the phantom and the tissue studies presented here indicate a change in the Rayleigh damping structure, described as Rayleigh composition, between different material types, with tofu and healthy tissue showing lower Rayleigh composition values than gelatin or cancerous tissue. CONCLUSIONS: It is possible that Rayleigh damping elastography and the concomitant Rayleigh composition images provide a mechanism for differentiating tissue structure in addition to measuring elastic stiffness and attenuation. Such information could be valuable in the use of Rayleigh damped magnetic resonance elastography as a diagnostic imaging tool. PMID- 21626933 TI - Elastic registration of multimodal prostate MRI and histology via multiattribute combined mutual information. AB - PURPOSE: By performing registration of preoperative multiprotocol in vivo magnetic resonance (MR) images of the prostate with corresponding whole-mount histology (WMH) sections from postoperative radical prostatectomy specimens, an accurate estimate of the spatial extent of prostate cancer (CaP) on in vivo MR imaging (MRI) can be retrospectively established. This could allow for definition of quantitative image-based disease signatures and lead to development of classifiers for disease detection on multiprotocol in vivo MRI. Automated registration of MR and WMH images of the prostate is complicated by dissimilar image intensities, acquisition artifacts, and nonlinear shape differences. METHODS: The authors present a method for automated elastic registration of multiprotocol in vivo MRI and WMH sections of the prostate. The method, multiattribute combined mutual information (MACMI), leverages all available multiprotocol image data to drive image registration using a multivariate formulation of mutual information. RESULTS: Elastic registration using the multivariate MI formulation is demonstrated for 150 corresponding sets of prostate images from 25 patient studies with T2-weighted and dynamic-contrast enhanced MRI and 85 image sets from 15 studies with an additional functional apparent diffusion coefficient MRI series. Qualitative results of MACMI evaluation via visual inspection suggest that an accurate delineation of CaP extent on MRI is obtained. Results of quantitative evaluation on 150 clinical and 20 synthetic image sets indicate improved registration accuracy using MACMI compared to conventional pairwise mutual information-based approaches. CONCLUSIONS: The authors' approach to the registration of in vivo multiprotocol MRI and ex vivo WMH of the prostate using MACMI is unique, in that (1) information from all available image protocols is utilized to drive the registration with histology, (2) no additional, intermediate ex vivo radiology or gross histology images need be obtained in addition to the routinely acquired in vivo MRI series, and (3) no corresponding anatomical landmarks are required to be identified manually or automatically on the images. PMID- 21626934 TI - Characterization of lamina propria and vocal muscle in human vocal fold tissue by ultrasound Nakagami imaging. AB - PURPOSE: A number of ultrasound techniques have been applied to identify the biomechanical properties of the vocal folds. These conventional ultrasound methods, however, are not capable of visually mapping the concentration of collagen and elastic fibers in the vocal folds in the form of a parametric image. This study proposes to use a statistical parameter, the Nakagami factor estimated from the statistical distribution of the ultrasonic signals backscattered from tissues, as a means for parametric imaging of the biomechanical properties of the vocal folds. METHODS: The ultrasonic backscattered signals were acquired from four larynges (eight vocal folds) obtained from individuals without vocal fold pathology for constructing the Nakagami images. The textures of the Nakagami image in the lamina propria (LP) and the vocal muscle (VM) were observed and compared. The average and standard deviation of the Nakagami parameter for the LP and the VM were also calculated. RESULTS: The results showed that the Nakagami parameter of the LP is larger than that of the VM. Moreover, the LP and the VM have different shading features in the Nakagami images. It was found that the Nakagami parameter may depend on the concentration of collagen and elastic fibers, demonstrating that the Nakagami imaging may allow visual differentiation between the LP and the VM in the vocal folds. CONCLUSIONS: Current preliminary results suggested that the high-frequency Nakagami imaging may allow real-time visual characterization of the vocal fold tissues in clinical routine examinations. PMID- 21626935 TI - Assessing software upgrades, plan properties and patient geometry using intensity modulated radiation therapy (IMRT) complexity metrics. AB - PURPOSE: The aim of this study is to compare the sensitivity of different metrics to detect differences in complexity of intensity modulated radiation therapy (IMRT) plans following upgrades, changes to planning parameters, and patient geometry. Correlations between complexity metrics are also assessed. METHOD: A program was developed to calculate a series of metrics used to describe the complexity of IMRT fields using monitor units (MUs) and multileaf collimator files: Modulation index (MI), modulation complexity score (MCS), and plan intensity map variation (PIMV). Each metric, including the MUs, was used to assess changes in beam complexity for six prostate patients, following upgrades in the inverse planning optimization software designed to incorporate direct aperture optimization (DAO). All beams were delivered to a 2D ionization chamber array and compared to those calculated using gamma analysis. Each complexity metric was then calculated for all beams, on a different set of six prostate IMRT patients, to assess differences between plans calculated using different minimum field sizes and different maximum segment numbers. Different geometries, including CShape, prostate, and head and neck phantoms, were also assessed using the metrics. Correlations between complexity metrics were calculated for 20 prostate IMRT patients. RESULTS: MU, MCS, MI, and PIMV could all detect reduced complexity following an upgrade to the optimization leaf sequencer, although only MI and MCS could detect a reduction in complexity when one-step optimization (DAO) was employed rather than two-step optimization. All metrics detected a reduction in complexity when the minimum field size was increased from 1 to 4 cm and all apart from PIMV detected reduced complexity when the number of segments was significantly reduced. All metrics apart from MI showed differences in complexity depending on the treatment site. Significant correlations exist between all metrics apart from MI and PIMV for prostate IMRT patients. Treatment deliverability appeared to be more correlated with MI and MCS than MU or PIMV. CONCLUSIONS: The application of complexity metrics in the IMRT treatment planning process has been demonstrated. Complexity of treatment plans can vary for different inverse planning software versions and can depend on planning parameters and the treatment site. MCS is most suitable for inclusion within the cost function to limit complexity in IMRT optimization due to its sensitivity to complexity changes and correlation to treatment deliverability. PMID- 21626936 TI - Performance of topological texture features to classify fibrotic interstitial lung disease patterns. AB - PURPOSE: Topological texture features were compared in their ability to classify "honeycombing," a morphological pattern that is considered indicative for the presence of fibrotic interstitial lung disease in high-resolution computed tomography (HRCT) images. METHODS: For 14 patients with known occurrence of honeycombing, a stack of 70 axial, lung kernel reconstructed images was acquired from HRCT chest exams. A set of 964 regions of interest of both healthy and pathological (356) lung tissue was identified by an experienced radiologist. Texture features were extracted using statistical features (Stat), six properties calculated from gray-level co-occurrence matrices (GLCMs), Minkowski dimensions (MDs), and three Minkowski functionals (MFs) (e.g., MF.Euler). A naive Bayes (NB) and k-nearest-neighbor (k-NN) classifier, a multilayer radial basis functions network (RBFN), and a support vector machine with a radial basis function (SVMrbf) kernel were optimized in a tenfold cross-validation for each texture vector, and the classification accuracy was calculated on independent test sets as a quantitative measure of automated tissue characterization. A Wilcoxon signed rank test was used to compare two accuracy distributions and the significance thresholds were adjusted for multiple comparisons by the Bonferroni correction. RESULTS: The best classification results were obtained by the MF features, which performed significantly better than all the standard Stat, GLCM, and MD features (p < 0.001) for both classifiers. The highest accuracies were found for MF.Euler (93.6%, 94.9%, 94.2%, and 95.0% for NB, k-NN, RBFN, and SVMrbf, respectively). The best groups of standard texture features were a Stat and GLCM ("homogeneity") feature set (up to 91.8%). CONCLUSIONS: The results indicate that advanced topological texture features derived from MFs can provide superior classification performance in computer-assisted diagnosis of fibrotic interstitial lung disease patterns when compared to standard texture analysis methods. PMID- 21626937 TI - Comparison of 250 MHz electron spin echo and continuous wave oxygen EPR imaging methods for in vivo applications. AB - PURPOSE: The authors compare two electron paramagnetic resonance imaging modalities at 250 MHz to determine advantages and disadvantages of those modalities for in vivo oxygen imaging. METHODS: Electron spin echo (ESE) and continuous wave (CW) methodologies were used to obtain three-dimensional images of a narrow linewidth, water soluble, nontoxic oxygen-sensitive trityl molecule OX063 in vitro and in vivo. The authors also examined sequential images obtained from the same animal injected intravenously with trityl spin probe to determine temporal stability of methodologies. RESULTS: A study of phantoms with different oxygen concentrations revealed a threefold advantage of the ESE methodology in terms of reduced imaging time and more precise oxygen resolution for samples with less than 70 torr oxygen partial pressure. Above 100 torr, CW performed better. The images produced by both methodologies showed pO2 distributions with similar mean values. However, ESE images demonstrated superior performance in low pO2 regions while missing voxels in high pO2 regions. CONCLUSIONS: ESE and CW have different areas of applicability. ESE is superior for hypoxia studies in tumors. PMID- 21626938 TI - Objective method to report planner-independent skin/rib maximal dose in balloon based high dose rate (HDR) brachytherapy for breast cancer. AB - PURPOSE: An objective method was proposed and compared with a manual selection method to determine planner-independent skin and rib maximal dose in balloon based high dose rate (HDR) brachytherapy planning. METHODS: The maximal dose to skin and rib was objectively extracted from a dose volume histogram (DVH) of skin and rib volumes. A virtual skin volume was produced by expanding the skin surface in three dimensions (3D) external to the breast with a certain thickness in the planning computed tomography (CT) images. Therefore, the maximal dose to this volume occurs on the skin surface the same with a conventional manual selection method. The rib was also delineated in the planning CT images and its maximal dose was extracted from its DVH. The absolute (Abdiff = [D(max) Man - D (max)DVH]) and relative (Rediff[%] = 100 x ([D(max)Man-D(max)DVH])/D(max)DVH) maximal skin and rib dose differences between the manual selection method (D(max)Man) and the objective method (D(max)DVH) were measured for 50 balloon based HDR (25 MammoSite and 25 Contura) patients. RESULTS: The average +/- standard deviation of maximal dose difference was 1.67% +/- 1.69% of the prescribed dose (PD). No statistical difference was observed between MammoSite and Contura patients for both Abdiff and Rediff[%] values. However, a statistically significant difference (p value < 0.0001) was observed in maximal rib dose difference compared with maximal skin dose difference for both Abdiff (2.30% +/- 1.71% vs 1.05% +/- 1.43%) and Rediff[%] (2.32% +/- 1.79% vs 1.21% +/- 1.41%). In general, rib has a more irregular contour and it is more proximally located to the balloon for 50 HDR patients. Due to the inverse square law factor, more dose difference was observed in higher dose range (D(max) > 90%) compared with lower dose range (D(max) < 90%): 2.16% +/- 1.93% vs 1.19% +/- 1.25% with p value of 0.0049. However, the Rediff[%] analysis eliminated the inverse square factor and there was no statistically significant difference (p value = 0.8931) between high and low dose ranges. CONCLUSIONS: The objective method using volumetric information of skin and rib can determine the planner-independent maximal dose compared with the manual selection method. However, the difference was < 2% of PD, on average, if appropriate attention is paid to selecting a manual dose point in 3D planning CT images. PMID- 21626939 TI - Correction for patient table-induced scattered radiation in cone-beam computed tomography (CBCT). AB - PURPOSE: In image-guided radiotherapy, an artifact typically seen in axial slices of x-ray cone-beam computed tomography (CBCT) reconstructions is a dark region or "black hole" situated below the scan isocenter. The authors trace the cause of the artifact to scattered radiation produced by radiotherapy patient tabletops and show it is linked to the use of the offset-detector acquisition mode to enlarge the imaging field-of-view. The authors present a hybrid scatter kernel superposition (SKS) algorithm to correct for scatter from both the object-of interest and the tabletop. METHODS: Monte Carlo simulations and phantom experiments were first performed to identify the source of the black hole artifact. For correction, a SKS algorithm was developed that uses separate kernels to estimate scatter from the patient tabletop and the object-of-interest. Each projection is divided into two regions, one defined by the shadow cast by the tabletop on the imager and one defined by the unshadowed region. The region not shadowed by the tabletop is processed using the recently developed fast adaptive scatter kernel superposition (fASKS) method which employs asymmetric kernels that best model scatter transport through bodylike objects. The shadowed region is convolved with a combination of slab-derived symmetric SKS kernels and asymmetric fASKS kernels. The composition of the hybrid kernels is projection angle-dependent. To test the algorithm, pelvis phantom and in vivo data were acquired using a CBCT test stand, a Varian Acuity simulator, and a Varian On Board Imager, all of which have similar geometries and components. Artifact intensities and Hounsfield unit (HU) accuracies in the reconstructions were assessed before and after the correction. RESULTS: The hybrid kernel algorithm provided effective correction and produced substantially better scatter estimates than the symmetric SKS or asymmetric fASKS methods alone. HU nonuniformities in the reconstructed pelvis phantom were reduced from 220 to 50 HU (i.e., 22%-5%). In the in vivo scans, the black hole artifact was reduced by up to 147 HU, a 73% improvement, and anatomical details in the prostate and rectum areas were made considerably more visible. CONCLUSIONS: Radiotherapy tabletops, which are generally flatter and larger than those used for diagnostic CT, can produce significant scatter-related artifacts. The proposed hybrid SKS algorithm accurately estimates scatter from both the object-of-interest and the patient tabletop, and resulting image uniformities and HU accuracies are greatly improved. PMID- 21626941 TI - Deformable planning CT to cone-beam CT image registration in head-and-neck cancer. AB - PURPOSE: The purpose of this work was to implement and validate a deformable CT to cone-beam computed tomography (CBCT) image registration method in head-and neck cancer to eventually facilitate automatic target delineation on CBCT. METHODS: Twelve head-and-neck cancer patients underwent a planning CT and weekly CBCT during the 5-7 week treatment period. The 12 planning CT images (moving images) of these patients were registered to their weekly CBCT images (fixed images) via the symmetric force Demons algorithm and using a multiresolution scheme. Histogram matching was used to compensate for the intensity difference between the two types of images. Using nine known anatomic points as registration targets, the accuracy of the registration was evaluated using the target registration error (TRE). In addition, region-of-interest (ROI) contours drawn on the planning CT were morphed to the CBCT images and the volume overlap index (VOI) between registered contours and manually delineated contours was evaluated. RESULTS: The mean TRE value of the nine target points was less than 3.0 mm, the slice thickness of the planning CT. Of the 369 target points evaluated for registration accuracy, the average TRE value was 2.6 +/- 0.6 mm. The mean TRE for bony tissue targets was 2.4 +/- 0.2 mm, while the mean TRE for soft tissue targets was 2.8 +/- 0.2 mm. The average VOI between the registered and manually delineated ROI contours was 76.2 +/- 4.6%, which is consistent with that reported in previous studies. CONCLUSIONS: The authors have implemented and validated a deformable image registration method to register planning CT images to weekly CBCT images in head-and-neck cancer cases. The accuracy of the TRE values suggests that they can be used as a promising tool for automatic target delineation on CBCT. PMID- 21626940 TI - Characterization and identification of spatial artifacts during 4D-CT imaging. AB - PURPOSE: The purpose of this work is twofold: First, to characterize the artifacts occurring in helical 4D-CT imaging; second, to propose a method that can automatically identify the artifacts in 4D-CT images. The authors have designed a process that can automatically identify the artifacts in 4D-CT images, which may be invaluable in quantifying the quality of 4D-CT images and reducing the artifacts from the reconstructed images on a large dataset. METHODS: Given two adjacent stacks obtained from the same respiration phase, the authors determine if there are artifacts between them. The proposed method uses a "bridge" stack strategy to connect the two stacks. Using normalized cross correlation convolution (NCCC), the two stacks are mapped to the bridge stack and the best matching positions can be located. Using this position information, the authors can then determine if there are artifacts between the two stacks. By combining the matching positions with NCCC values, the performance can be improved. RESULTS: To validate the method, three expert observers independently labeled over 600 stacks on five patients. The results confirmed that high performance was obtained using the proposed method. The average sensitivity was about 0.87 and the average specificity was 0.82. The proposed method also outperformed the method of using respiratory signal (sensitivity increased from 0.50 to 0.87 and specificity increased from 0.70 to 0.82). CONCLUSIONS: This study shows that the spatial artifacts during 4D-CT imaging are characterized and can be located automatically by the proposed method. The method is relatively simple but effective. It provides a way to evaluate the artifacts more objectively and accurately. The reported approach has promising potential for automatically identifying the types and frequency of artifacts on large scale 4D CT image dataset. PMID- 21626942 TI - Oblique incidence effects in direct x-ray detectors: a first-order approximation using a physics-based analytical model. AB - PURPOSE: The authors describe the modifications to a previously developed analytical model of indirect CsI:Tl-based detector response required for studying oblique x-ray incidence effects in direct semiconductor-based detectors. This first-order approximation analysis allows the authors to describe the associated degradation in resolution in direct detectors and compare the predictions to the published data for indirect detectors. METHODS: The proposed model is based on a physics-based analytical description developed by Freed et al. ["A fast, angle dependent, analytical model of CsI detector response for optimization of 3D x-ray breast imaging systems," Med. Phys. 37(6), 2593-2605 (2010)] that describes detector response functions for indirect detectors and oblique incident x rays. The model, modified in this work to address direct detector response, describes the dependence of the response with x-ray energy, thickness of the transducer layer, and the depth-dependent blur and collection efficiency. RESULTS: The authors report the detector response functions for indirect and direct detector models for typical thicknesses utilized in clinical systems for full-field digital mammography (150 microm for indirect CsI:Tl and 200 microm for a-Se direct detectors). The results suggest that the oblique incidence effect in a semiconductor detector differs from that in indirect detectors in two ways: The direct detector model produces a sharper overall PRF compared to the response corresponding to the indirect detector model for normal x-ray incidence and a larger relative increase in blur along the x-ray incidence direction compared to that found in indirect detectors with respect to the response at normal incidence angles. CONCLUSIONS: Compared to the effect seen in indirect detectors, the direct detector model exhibits a sharper response at normal x-ray incidence and a larger relative increase in blur along the x-ray incidence direction with respect to the blur in the orthogonal direction. The results suggest that the oblique incidence effect in direct detectors can be considered to be caused mostly by the geometry of the path where the x-ray beam and its secondary particles deposit energy in the semiconductor layer. PMID- 21626943 TI - Method for measuring the focal spot size of an x-ray tube using a coded aperture mask and a digital detector. AB - PURPOSE: The goal of this study is to evaluate a new method based on a coded aperture mask combined with a digital x-ray imaging detector for measurements of the focal spot sizes of diagnostic x-ray tubes. Common techniques for focal spot size measurements employ a pinhole camera, a slit camera, or a star resolution pattern. The coded aperture mask is a radiation collimator consisting of a large number of apertures disposed on a predetermined grid in an array, through which the radiation source is imaged onto a digital x-ray detector. The method of the coded mask camera allows one to obtain a one-shot accurate and direct measurement of the two dimensions of the focal spot (like that for a pinhole camera) but at a low tube loading (like that for a slit camera). A large number of small apertures in the coded mask operate as a "multipinhole" with greater efficiency than a single pinhole, but keeping the resolution of a single pinhole. METHODS: X-ray images result from the multiplexed output on the detector image plane of such a multiple aperture array, and the image of the source is digitally reconstructed with a deconvolution algorithm. Images of the focal spot of a laboratory x-ray tube (W anode: 35-80 kVp; focal spot size of 0.04 mm) were acquired at different geometrical magnifications with two different types of digital detector (a photon counting hybrid silicon pixel detector with 0.055 mm pitch and a flat panel CMOS digital detector with 0.05 mm pitch) using a high resolution coded mask (type no two-holes-touching modified uniformly redundant array) with 480 0.07 mm apertures, designed for imaging at energies below 35 keV. Measurements with a slit camera were performed for comparison. A test with a pinhole camera and with the coded mask on a computed radiography mammography unit with 0.3 mm focal spot was also carried out. RESULTS: The full width at half maximum focal spot sizes were obtained from the line profiles of the decoded images, showing a focal spot of 0.120 mm x 0.105 mm at 35 kVp and M = 6.1, with a detector entrance exposure as low as 1.82 mR (0.125 mA s tube load). The slit camera indicated a focal spot of 0.112 mm x 0.104 mm at 35 kVp and M = 3.15, with an exposure at the detector of 72 mR. Focal spot measurements with the coded mask could be performed up to 80 kVp. Tolerance to angular misalignment with the reference beam up to 7 degrees in in-plane rotations and 1 degrees deg in out-of-plane rotations was observed. The axial distance of the focal spot from the coded mask could also be determined. It is possible to determine the beam intensity via measurement of the intensity of the decoded image of the focal spot and via a calibration procedure. CONCLUSIONS: Coded aperture masks coupled to a digital area detector produce precise determinations of the focal spot of an x-ray tube with reduced tube loading and measurement time, coupled to a large tolerance in the alignment of the mask. PMID- 21626944 TI - Crescent artifacts in cone-beam CT. AB - PURPOSE: In image-guided radiation therapy, cone-beam CT has been adopted for three-dimensional target localization in the treatment room. In many of these cone-beam CT images, dark and light crescent artifacts can be seen. This study investigates potential causes of this artifact and a technique for mitigating the crescents. METHODS: Three deviations from an ideal geometry were simulated to assess their ability to cause crescent artifacts: Bowtie filter sag, x-ray tube sag, and x-ray tube rotation. The magnitudes of these deviations were estimated by matching shifts in simulated projections to those observed with clinical systems. To correct the artifacts, angle-dependent blank projections were acquired and incorporated into image reconstruction. The degree of artifact reduction was evaluated with varying numbers (1-380) of blank projections. Scanner-acquired phantom and patient studies were conducted to demonstrate the effectiveness of the proposed correction method. RESULTS: All three investigated causes of the crescent artifact introduced similar mismodeling of the acquired projections and similar crescent artifacts. The deviations required for these artifacts were in the range of 0.5-5 mm or 0.1 degrees. RMS error is reduced from 8.91 x 10(-4) to 5.25 x 10(-7) for 1-380 blank projections over a 200 degrees scan angle. In the patient and phantom studies, reconstructions that utilized 380 blank projections largely mitigated the crescent artifacts. CONCLUSIONS: Small deviations from an ideal geometry can result in crescent artifacts due to steep gradients in the bowtie filter. Angle-dependent blank projections can largely alleviate the artifacts. PMID- 21626945 TI - Local and global 3D noise power spectrum in cone-beam CT system with FDK reconstruction. AB - PURPOSE: The authors examine the nonstationary noise behavior of a cone-beam CT system with FDK reconstruction. METHODS: To investigate the nonstationary noise behavior, an analytical expression for the NPS of local volumes and an entire volume was derived and quantitatively compared to the NPS estimated from experimental air and water images. RESULTS: The NPS of local volumes at different locations along the z-axis showed radial symmetry in the f(x)-f(y) plane and different missing cone regions in the f(z) direction depending on the tilt angle of rays through the local volumes. For local volumes away from the z-axis, the NPS of air and water images showed sharp transitions in the f(x)-f(y) and f(y) f(z) planes and lack of radial symmetry in the f(x)-f(y) plane. These effects are mainly caused by varying magnification and different noise levels from view to view. In the NPS of the entire volume, the f(x)-f(y) plane showed radial symmetry because the nonstationary noise behaviors of local volumes were averaged out. The nonstationary sharp transitions were manifested as a high-frequency roll-off. CONCLUSIONS: The results from noise power analysis for local volumes and an entire volume demonstrate the spatially varying noise behavior in the reconstructed cone-beam CT images. PMID- 21626946 TI - Evaluation of the quantitative accuracy of a commercially available positron emission mammography scanner. AB - PURPOSE: The PEM Flex Solo II (Naviscan, Inc., San Diego, CA) is currently the only commercially available positron emission mammography (PEM) scanner. This scanner does not apply corrections for count rate effects, attenuation, or scatter during image reconstruction, potentially affecting the quantitative accuracy of images. The aim of this work is to measure the overall quantitative accuracy of the PEM Flex system and to determine the individual contributions of error from count rate effects, attenuation, and scatter. METHODS: Gelatin phantoms were designed to simulate breasts of different thicknesses (4-12 cm) with varying uniform background activity concentration (AC) (0.007-0.5 microCi/cc), cysts, and lesions (2:1, 5:1, and 10:1 lesion-to-background ratios). The overall error was calculated from ROI measurements in the phantoms with a clinically relevant background AC (0.065 microCi/cc). The error due to count rate effects was determined by comparing the overall error at multiple background AC to the error at a very low background AC (0.007 microCi/cc) where count rate effects are considered negligible. A point source and cold gelatin phantoms of different thicknesses were used to assess the errors due to attenuation and scatter. The maximum pixel values of the point source in gelatin and in air were compared to determine the effect of attenuation, while scatter was evaluated by comparing the sum of all pixel values in gelatin and in air. RESULTS: The AC in the uniform background was underestimated in phantoms of all thicknesses, with the exception of the 4-cm-thick phantoms in which the measured AC was within +/- 7% of the true value (0.065 microCi/cc). The degree of underestimation in the uniform background increased with the phantom thickness, up to 34% +/- 6% in the 12 cm phantoms. The AC in all lesions was underestimated by more than that measured in the background (22% for the 2:1 lesions in the 4 cm phantom) and this underestimation increased with increasing thickness and lesion-to-background ratio (85% for the 10:1 lesions in the 12 cm phantoms). The error due to count rate effects reduced the measured AC with respect to its true value. This error increased with increasing background AC (23% +/- 6% at 0.065 microCi/cc to 85% +/ 1% at 0.5 microCi/cc for the 4 cm phantoms) and decreased with increasing phantom thickness (23% +/- 6% for the 4-cm-thick phantoms to 7% +/- 7% for the 12 cm-thick phantoms at 0.065 microCi/cc). Attenuation was a substantial source of error and reduced the measured AC by 51% +/- 10%-77% +/- 4% in the 4-12 cm phantoms, respectively. Scatter increased the total signal measured in images by a relatively constant amount (23% +/- 9%) for all thicknesses. CONCLUSIONS: Applying corrections for count rate effects, attenuation, and scatter will be essential for the PEM Flex Solo II to be able to produce quantitatively accurate images. PMID- 21626948 TI - A method for robust segmentation of arbitrarily shaped radiopaque structures in cone-beam CT projections. AB - PURPOSE: Implanted markers are commonly used in radiotherapy for x-ray based target localization. The projected marker position in a series of cone-beam CT (CBCT) projections can be used to estimate the three dimensional (3D) target trajectory during the CBCT acquisition. This has important applications in tumor motion management such as motion inclusive, gating, and tumor tracking strategies. However, for irregularly shaped markers, reliable segmentation is challenged by large variations in the marker shape with projection angle. The purpose of this study was to develop a semiautomated method for robust and reliable segmentation of arbitrarily shaped radiopaque markers in CBCT projections. METHODS: The segmentation method involved the following three steps: (1) Threshold based segmentation of the marker in three to six selected projections with large angular separation, good marker contrast, and uniform background; (2) construction of a 3D marker model by coalignment and backprojection of the threshold-based segmentations; and (3) construction of marker templates at all imaging angles by projection of the 3D model and use of these templates for template-based segmentation. The versatility of the segmentation method was demonstrated by segmentation of the following structures in the projections from two clinical CBCT scans: (1) Three linear fiducial markers (Visicoil) implanted in or near a lung tumor and (2) an artificial cardiac valve in a lung cancer patient. RESULTS: Automatic marker segmentation was obtained in more than 99.9% of the cases. The segmentation failed in a few cases where the marker was either close to a structure of similar appearance or hidden behind a dense structure (data cable). CONCLUSIONS: A robust template based method for segmentation of arbitrarily shaped radiopaque markers in CBCT projections was developed. PMID- 21626947 TI - Spectral method for the correction of the Cerenkov light effect in plastic scintillation detectors: a comparison study of calibration procedures and validation in Cerenkov light-dominated situations. AB - PURPOSE: The purposes of this work were: (1) To determine if a spectral method can accurately correct the Cerenkov light effect in plastic scintillation detectors (PSDs) for situations where the Cerenkov light is dominant over the scintillation light and (2) to develop a procedural guideline for accurately determining the calibration factors of PSDs. METHODS: The authors demonstrate, by using the equations of the spectral method, that the condition for accurately correcting the effect of Cerenkov light is that the ratio of the two calibration factors must be equal to the ratio of the Cerenkov light measured within the two different spectral regions used for analysis. Based on this proof, the authors propose two new procedures to determine the calibration factors of PSDs, which were designed to respect this condition. A PSD that consists of a cylindrical polystyrene scintillating fiber (1.6 mm3) coupled to a plastic optical fiber was calibrated by using these new procedures and the two reference procedures described in the literature. To validate the extracted calibration factors, relative dose profiles and output factors for a 6 MV photon beam from a medical linac were measured with the PSD and an ionization chamber. Emphasis was placed on situations where the Cerenkov light is dominant over the scintillation light and on situations dissimilar to the calibration conditions. RESULTS: The authors found that the accuracy of the spectral method depends on the procedure used to determine the calibration factors of the PSD and on the attenuation properties of the optical fiber used. The results from the relative dose profile measurements showed that the spectral method can correct the Cerenkov light effect with an accuracy level of 1%. The results obtained also indicate that PSDs measure output factors that are lower than those measured with ionization chambers for square field sizes larger than 25 x 25 cm2, in general agreement with previously published Monte Carlo results. CONCLUSIONS: The authors conclude that the spectral method can be used to accurately correct the Cerenkov light effect in PSDs. The authors confirmed the importance of maximizing the difference of Cerenkov light production between calibration measurements. The authors also found that the attenuation of the optical fiber, which is assumed to be constant in the original formulation of the spectral method, may cause a variation of the calibration factors in some experimental setups. PMID- 21626950 TI - Effective and equivalent organ doses in patients undergoing coronary angiography and percutaneous coronary intervention. AB - PURPOSE: Recent recommendations of the International Commission on Radiological Protection state that the use of effective dose (E) for assessing patient exposure has severe limitations, though it can be kept for dose comparisons. In cardiology procedures, the equivalent dose (H(T)) is one of the most appropriate dose quantity to be evaluated for risk-benefit assessment. In this study, both E and H(T) values for ten critical organs in coronary angiography (CA) and percutaneous coronary interventions (PCI) were derived from in-the-field dose area-product (DAP) measurements in order to provide a database for doses in those procedures. METHODS: Conversion factors E/DAP calculated by Monte Carlo methods in two different mathematical human phantoms were applied to DAP values measured on 193 patients (118 CA and 75 PCI). Partial DAP values were recorded in-the field for each projection and for all patients. The partial effective doses of all projections were summed up to calculate the E of the entire procedure. Similarly, equivalent doses for ten critical organs/tissues (bone, colon, heart, liver, lung, esophagus, red bone marrow, skin, stomach, and thyroid) were derived from H(T)/DAP conversion factors for different projections calculated by Monte Carlo method. RESULTS: All parameters related to the patient dose, i.e., fluoroscopy times, number of images, DAP, effective doses, and equivalent doses, show a wide range of values depending on the complexity of the patient case and the experience of the cardiologist. The mean fluoroscopy time, DAP, and E values for coronary angiography patients were approximately threefold lower than those for PCI patients; the number of images for CA was half that for PCI. The correlation between effective dose and DAP was excellent for both CA and PCI. The equivalent doses values were in good correlations with DAP values in CA examinations, with Pearson's coefficients ranging from 0.87 (stomach) to 0.99 (skin) and r(mean) = 0.94. The same analysis was performed for PCI procedures. In this case, the trends were only slightly worse because "r" ranged from 0.70 (stomach) to 0.92 (bone) and r(mean) = 0.85. Simple conversion coefficients to estimate equivalent doses to ten critical organs/tissues from DAP values, for both CA and PCI, were provided for avoiding the need to carry out detailed in-the field analysis for all projections and for all patients. CONCLUSIONS: Measurements in-the-field of DAP values were carried out for two common cardiology procedures and effective doses were derived for each technique from detailed analysis of dose and projection data, using conversion factors provided by two different theoretical models. Equivalent doses to organs/tissues were also calculated using conversion factors proposed in the literature for different projections and cumulative conversion factors (H)T/DAP for ten organs/tissues were estimated. PMID- 21626949 TI - Nonconvex prior image constrained compressed sensing (NCPICCS): theory and simulations on perfusion CT. AB - PURPOSE: To present and evaluate a new image reconstruction method for dynamic CT based on a nonconvex prior image constrained compressed sensing (NCPICCS) algorithm. The authors systematically compared the undersampling potential, functional information recovery, and solution convergence speed of four compressed sensing (CS) based image reconstruction methods using perfusion CT data: Standard l1-based CS, nonconvex CS (NCCS), and l1-based and nonconvex CS, including an additional constraint based on a prior image (PICCS and NCPICCS, respectively). METHODS: The Shepp-Logan phantom was modified such that its uppermost ellipses changed attenuation through time, simulating both an arterial input function (AIF) and a homogeneous tissue perfusion region. Data were simulated with and without Poisson noise added to the projection data and subsequently reconstructed with all four CS-based methods at four levels of undersampling: 20, 12, 6, and 4 projections. Root mean squared (RMS) error of reconstructed images and recovered time attenuation curves (TACs) were assessed as well as convergence speed. The performance of both PICCS and NCPICCS methods were also evaluated using a kidney perfusion animal experiment data set. RESULTS: All four CS-based methods were able to reconstruct the phantoms with 20 projections, with similar results on the RMS error of the recovered TACs. NCCS allowed accurate reconstructions with as few as 12 projections, PICCS with as few as six projections, and NCPICCS with as few as four projections. These results were consistent for noise-free and noisy data. NCPICCS required the fewest iterations to converge across all simulation conditions, followed by PICCS, NCCS, and then CS. On animal data, at the lowest level of undersampling tested (16 projections), the image quality of NCPICCS was better than PICCS with fewer streaking artifacts, while the TAC accuracy on the selected region of interest was comparable. CONCLUSIONS: The authors have presented a novel method for image reconstruction using highly undersampled dynamic CT data. The NCPICCS method takes advantage of the information provided by a prior image, as in PICCS, but employs a more general nonconvex sparsity measure [such as the l(p)-norm (0 < p < or = 1)] rather than the conventional convex l1-norm. Despite the lack of guarantees of a globally optimal solution, the proposed nonconvex extension of PICCS consistently allowed for image reconstruction from fewer samples than the analogous l1-based PICCS method. Both nonconvex sparsity measures as well as prior image information (when available) significantly reduced the number of iterations required for convergence, potentially providing computational advantages for practical implementation of CS-based image reconstruction techniques. PMID- 21626951 TI - Technical note: removing the stem effect when performing Ir-192 HDR brachytherapy in vivo dosimetry using plastic scintillation detectors: a relevant and necessary step. AB - PURPOSE: The purpose of this study was to investigate whether or not a stem effect removal technique is necessary when performing Ir-192 HDR brachytherapy in vivo dosimetry using a scintillation detector. METHODS: A red-green-blue photodiode connected to a multichannel electrometer was used to detect the light emitted from a plastic scintillation detector (PSD) during irradiation with an Ir 192 HDR brachytherapy source. Accuracy in dose measurement was compared with and without the use of stem effect removal techniques. Monochromatic and polychromatic filtration techniques were studied. An in-house template was built for accurate positioning of catheters in which the source and the PSD were inserted. Dose distribution was measured up to 5 cm from source to detector in the radial and longitudinal directions. RESULTS: The authors found the stem effect to be particularly important when the source was close to the optical fiber guide and far from the scintillation component of the detector. It can account for up to (72 +/- 3)% of the signal under clinically relevant conditions. The polychromatic filtration outperformed the monochromatic filtration as well as the absence of filtration in regard to dose measurement accuracy. CONCLUSIONS: It is necessary to implement a stem effect removal technique when building a PSD for in vivo dosimetry during Ir-192 HDR brachytherapy. The PSD that the authors have developed for this study would be suitable for such an application. PMID- 21626952 TI - The characterization of breast anatomical metrics using dedicated breast CT. AB - PURPOSE: Accurate anatomical characterization of the breast is useful in breast phantom development and computer modeling of breast imaging technologies. Capitalizing on the three-dimensional capabilities of dedicated breast CT (bCT), a number of parameters which describe breast shape and fibroglandular distribution are defined. METHODS: Among 219 bCT data sets, the effective diameter and length of the pendant breast as well as the breast volume were measured and characterized for each bra cup size. The volume glandular fraction (VGF) was determined as a function of patient age, BIRADS density, bra cup size, and breast diameter. The glandular fraction was examined in coronal and sagittal planes of the breast, and the radial distribution of breast glandular fraction within a coronal bCT image was examined for three breast regions. The areal glandular fraction (AGF) was estimated from two-dimensional projections of the breast (simulated by projecting bCT data sets) and was compared to the corresponding VGF. RESULTS: The effective breast diameter and length increase with increasing bra cup size. The mean breast diameters (+/- standard error) of bra cup sizes A/AA, B, C, and D/DD were 11.1 +/- 0.5, 11.4 +/- 0.3, 13.0 +/- 0.2, and 13.7 +/- 0.2 cm, respectively. VGF was lower among older women and those with larger breast diameter and larger bra cup size. VGF increased as a function of the reported BIRADS density. AGF increased with VGF. Fibroglandular tissue was distributed primarily in the central portion of the breast. CONCLUSIONS: Breast metrics were examined and a number of parameters were defined which may be useful for breast modeling. The reported data may provide researchers with useful information for characterizing the breast for various imaging or dosimetry tasks. PMID- 21626953 TI - Collimator design for experimental minibeam radiation therapy. AB - PURPOSE: To design and optimize a minibeam collimator for minibeam radiation therapy studies using a 250 kVp x-ray machine as a simulated synchrotron source. METHODS: A Philips RT250 orthovoltage x-ray machine was modeled using the EGSnrc/BEAMnrc Monte Carlo software. The resulting machine model was coupled to a model of a minibeam collimator with a beam aperture of 1 mm. Interaperture spacing and collimator thickness were varied to produce a minibeam with the desired peak-to-valley ratio. RESULTS: Proper design of a minibeam collimator with Monte Carlo methods requires detailed knowledge of the x-ray source setup. For a cathode-ray tube source, the beam spot size, target angle, and source shielding all determine the final valley-to-peak dose ratio. CONCLUSIONS: A minibeam collimator setup was created, which can deliver a 30 Gy peak dose minibeam radiation therapy treatment at depths less than 1 cm with a valley-to peak dose ratio on the order of 23%. PMID- 21626954 TI - Classification of scattering media within benign and malignant breast tumors based on ultrasound texture-feature-based and Nakagami-parameter images. AB - PURPOSE: Benign and malignant tumors can be classified by using texture analysis of the ultrasound B-scan image to describe the variation in the echogenicity of scatterers. The recently proposed ultrasonic Nakagami parametric image has also been used to detect the concentrations and arrangements of scatterers for tumor characterization applications. B-scan-based texture analysis and the Nakagami parametric image are functionally complementary in ultrasonic tissue characterizations and this study aimed to combine these methods in order to improve the ability to characterize breast tumors. METHODS: To validate this concept, radio-frequency data obtained from 130 clinical cases were used to construct the texture-feature parametric image and the Nakagami parametric image. Four texture-feature parameters based on a gray-level co-occurrence matrix (homogeneity, contrast, energy, and variance) and the Nakagami parameters of the benign and malignant tumors were calculated. The usefulness of an individual parameter was determined and scatter graphs indicated the relationship between two selected texture-feature parameters. Fisher's linear discriminant analysis was used to combine the selected texture-feature parameters with the Nakagami parameter. The performance in classifying tumors was evaluated based on the receiver operating characteristic curve. RESULTS: The results indicated that there is a trade-off between sensitivity and specificity when using an individual texture-feature parameter or when combining two such correlated parameters to discriminate benign and malignant cases. However, the best performance was obtained when combining selected texture-feature parameters with the Nakagami parameter. CONCLUSIONS: The study findings suggest that combining B-scan-based texture analysis and the Nakagami parametric image could improve the ability to classify benign and malignant breast tumors. PMID- 21626955 TI - Dosimetric validation of Acuros XB with Monte Carlo methods for photon dose calculations. AB - PURPOSE: The dosimetric accuracy of the recently released Acuros XB advanced dose calculation algorithm (Varian Medical Systems, Palo Alto, CA) is investigated for single radiation fields incident on homogeneous and heterogeneous geometries, and a comparison is made to the analytical anisotropic algorithm (AAA). METHODS: Ion chamber measurements for the 6 and 18 MV beams within a range of field sizes (from 4.0 x 4.0 to 30.0 x 30.0 cm2) are used to validate Acuros XB dose calculations within a unit density phantom. The dosimetric accuracy of Acuros XB in the presence of lung, low-density lung, air, and bone is determined using BEAMnrc/DOSXYZnrc calculations as a benchmark. Calculations using the AAA are included for reference to a current superposition/convolution standard. RESULTS: Basic open field tests in a homogeneous phantom reveal an Acuros XB agreement with measurement to within +/- 1.9% in the inner field region for all field sizes and energies. Calculations on a heterogeneous interface phantom were found to agree with Monte Carlo calculations to within +/- 2.0% (sigmaMC = 0.8%) in lung (p = 0.24 g cm(-3)) and within +/- 2.9% (sigmaMC = 0.8%) in low-density lung (p = 0.1 g cm(-3)). In comparison, differences of up to 10.2% and 17.5% in lung and low-density lung were observed in the equivalent AAA calculations. Acuros XB dose calculations performed on a phantom containing an air cavity (p = 0.001 g cm(-3)) were found to be within the range of +/- 1.5% to +/- 4.5% of the BEAMnrc/DOSXYZnrc calculated benchmark (sigmaMC = 0.8%) in the tissue above and below the air cavity. A comparison of Acuros XB dose calculations performed on a lung CT dataset with a BEAMnrc/DOSXYZnrc benchmark shows agreement within +/- 2%/2mm and indicates that the remaining differences are primarily a result of differences in physical material assignments within a CT dataset. CONCLUSIONS: By considering the fundamental particle interactions in matter based on theoretical interaction cross sections, the Acuros XB algorithm is capable of modeling radiotherapy dose deposition with accuracy only previously achievable with Monte Carlo techniques. PMID- 21626956 TI - Accuracies of the synthesized monochromatic CT numbers and effective atomic numbers obtained with a rapid kVp switching dual energy CT scanner. AB - PURPOSE: This study was performed to investigate the accuracies of the synthesized monochromatic images and effective atomic number maps obtained with the new GE Discovery CT750 HD CT scanner. METHODS: A Gammex-RMI model 467 tissue characterization phantom and the CT number linearity section of a Phantom Laboratory Catphan 600 phantom were scanned using the dual energy (DE) feature on the GE CT750 HD scanner. Synthesized monochromatic images at various energies between 40 and 120 keV and effective atomic number (Z(eff)) maps were generated. Regions of interest were placed within these images/maps to measure the average monochromatic CT numbers and average Z(eff) of the materials within these phantoms. The true Z(eff) values were either supplied by the phantom manufacturer or computed using Mayneord's equation. The linear attenuation coefficients for the true CT numbers were computed using the NIST XCOM program with the input of manufacturer supplied elemental compositions and densities. The effects of small variations in the assumed true densities of the materials were also investigated. Finally, the effect of body size on the accuracies of the synthesized monochromatic CT numbers was investigated using a custom lumbar section phantom with and without an external fat-mimicking ring. RESULTS: Other than the Z(eff) of the simulated lung inserts in the tissue characterization phantom, which could not be measured by DECT, the Z(eff) values of all of the other materials in the tissue characterization and Catphan phantoms were accurate to 15%. The accuracies of the synthesized monochromatic CT numbers of the materials in both phantoms varied with energy and material. For the 40-120 keV range, RMS errors between the measured and true CT numbers in the Catphan are 8-25 HU when the true CT numbers were computed using the nominal plastic densities. These RMS errors improve to 3 12 HU for assumed true densities within the nominal density +/- 0.02 g/cc range. The RMS errors between the measured and true CT numbers of the tissue mimicking materials in the tissue characterization phantom over the 40-120 keV range varied from about 6 HU-248 HU and did not improve as dramatically with small changes in assumed true density. CONCLUSIONS: Initial tests indicate that the Z(eff) values computed with DECT on this scanner are reasonably accurate; however, the synthesized monochromatic CT numbers can be very inaccurate, especially for dense tissue mimicking materials at low energies. Furthermore, the synthesized monochromatic CT numbers of materials still depend on the amount of the surrounding tissues especially at low keV, demonstrating that the numbers are not truly monochromatic. Further research is needed to develop DE methods that produce more accurate synthesized monochromatic CT numbers. PMID- 21626957 TI - Empirical binary tomography calibration (EBTC) for the precorrection of beam hardening and scatter for flat panel CT. AB - PURPOSE: Scatter and beam hardening are prominent artifacts in x-ray CT. Currently, there is no precorrection method that inherently accounts for tube voltage modulation and shaped prefiltration. METHODS: A method for self calibration based on binary tomography of homogeneous objects, which was proposed by B. Li et al. ["A novel beam hardening correction method for computed tomography," in Proceedings of the IEEE/ICME International Conference on Complex Medical Engineering CME 2007, pp. 891-895, 23-27 May 2007], has been generalized in order to use this information to preprocess scans of other, nonbinary objects, e.g., to reduce artifacts in medical CT applications. Further on, the method was extended to handle scatter besides beam hardening and to allow for detector pixel specific and ray-specific precorrections. This implies that the empirical binary tomography calibration (EBTC) technique is sensitive to spectral effects as they are induced by the heel effect, by shaped prefiltration, or by scanners with tube voltage modulation. The presented method models the beam hardening correction by using a rational function, while the scatter component is modeled using the pep model of B. Ohnesorge et al. ["Efficient object scatter correction algorithm for third and fourth generation CT scanners," Eur. Radiol. 9(3), 563-569 (1999)]. A smoothness constraint is applied to the parameter space to regularize the underdetermined system of nonlinear equations. The parameters determined are then used to precorrect CT scans. RESULTS: EBTC was evaluated using simulated data of a flat panel cone-beam CT scanner with tube voltage modulation and bow-tie prefiltration and using real data of a flat panel cone-beam CT scanner. In simulation studies, where the ground truth is known, the authors' correction model proved to be highly accurate and was able to reduce beam hardening by 97% and scatter by about 75%. Reconstructions of measured data showed significantly less artifacts than the standard reconstruction. CONCLUSIONS: EBTC appears to be an efficient algorithm to precorrect CT raw data for beam hardening and scatter and it can account for ray-dependent spectral variations as they occur due to the heel effect, due to shaped prefiltration, or due to variations in tube voltage. PMID- 21626958 TI - Technical note: unsupervised C-arm pose tracking with radiographic fiducial. AB - PURPOSE: C-arm fluoroscopy reconstruction, such as that used in prostate brachytherapy, requires that the relative poses of the individual C-arm fluoroscopy images must be known prior to reconstruction. Radiographic fiducials can provide excellent C-arm pose tracking, but they need to be segmented in the image. The authors report an automated and unsupervised method that does not require prior segmentation of the fiducial. METHODS: The authors compute the individual C-arm poses relative to a stationary radiographic fiducial of known geometry. The authors register a filtered 2D fluoroscopy image of the fiducial to its 3D model by using image intensity alone without prior segmentation. To enhance the C-arm images, the authors investigated a three-step cascade filter and a line enhancement filter. The authors tested the method on a composite fiducial containing beads, straight lines, and ellipses. Ground-truth C-arm pose was provided by a clinically proven method. RESULTS: Using 111 clinical C-arm images and +/- 10 degrees and +/- 10 mm random perturbation around the ground truth pose, a total of 2775 cases were evaluated. The average rotation and translation errors were 0.62 degrees (STD = 0.31 degrees) and 0.72 mm (STD = 0.55 mm) for the three-step filter and 0.67 degrees (STD = 0.40 degrees) and 0.87 mm (STD = 0.27 mm) using the line enhancement filter. CONCLUSIONS: The C-arm pose tracking method was sufficiently accurate and robust on human patient data for subsequent 3D implant reconstruction. PMID- 21626959 TI - On the use of the MLC dosimetric leaf gap as a quality control tool for accurate dynamic IMRT delivery. AB - PURPOSE: MLC leaf gap consistency is critical for the accurate delivery of dynamic IMRT plans. It is estimated that a systematic MLC leaf gap change of 0.6 mm will result in a 2% change to the equivalent uniform dose to a clinical target volume for a typical head and neck sliding window (SW) IMRT plan. The aim of this work is to use the measured dosimetric leaf gap (DLG) to verify the dosimetric reproducibility of dynamically delivered SW IMRT plans. This study focuses on Varian linacs equipped with the 120 Millennium MLC and the Eclipse treatment planning system (TPS), but can be extended to other linac/MLC/TPS combination. METHODS: An ionization chamber, a diode array, and an electronic portal imaging device (EPID) were used to assess the DLG in zero (central axis), one, and two dimensions, respectively. The DLG for zero and two dimensions was derived from measurements of SW fields of decreasing width (2, 1.5, 1, and 0.5 cm). The DLG in one dimension was measured directly from a single SW sweeping across a linear diode array. This one-dimensional DLG measurement was based on the full width at half maximum (FWHM) of the dose rate versus time spectrum. RESULTS: The DLG derived from ion chamber measurements at central axis agrees to within 0.1 mm, with the DLG measured directly from the FWHM of dose rate versus time spectrum. The measured DLG depends on the control points used for the MLC SW fields. When two control points were used, the DLG measured at central axis showed an increase of 0.6 mm with respect to the same measurements performed using three or more control points. The two-dimensional distribution of DLG obtained using the EPID identified leaf gap errors as small as +/- 0.2 mm in isolated areas away from central axis. CONCLUSIONS: Comprehensive measurements of the DLG in 0D, 1D, and 2D provide an accurate assessment of DLG value required during TPS commissioning. These DLG measurements can also be used as a quality control tool to quantify changes of the MLC calibration and leaf gap consistency, which is critical for the accurate delivery of dynamically delivered SW IMRT plans. PMID- 21626960 TI - Independent quality assurance of a helical tomotherapy machine using the dose magnifying glass. AB - PURPOSE: Helical tomotherapy is a complex delivery technique, integrating CT image guidance and intensity modulated radiotherapy in a single system. The integration of the CT detector ring on the gantry not only allows patient position verification but is also often used to perform various QA procedures. This convenience lacks the rigor of a machine-independent QA process. METHODS: In this article, a Si strip detector, known as the Dose Magnifying Glass (DMG), was used to perform machine-independent QA measurements of the multileaf collimator alignment, leaf open time threshold, and leaf fluence output factor (LFOF). RESULTS: The DMG measurements showed good agreements with EDR2 film for the MLC alignment test while the CT detector agrees well with DMG measurements for leaf open time threshold and LFOF measurements. The leaf open time threshold was found to be approximately 20 ms. The LFOF measured with the DMG agreed within error with the CT detector measured LFOF. CONCLUSIONS: The DMG with its 0.2 mm spatial resolution coupled to TERA ASIC allowed real-time high temporal resolution measurements of the tomotherapy leaf movement. In conclusion, DMG was shown to be a suitable tool for machine-independent QA of a tomotherapy unit. PMID- 21626961 TI - Investigation of radiological properties and water equivalency of PRESAGE dosimeters. AB - PURPOSE: PRESAGE is a dosimeter made of polyurethane, which is suitable for 3D dosimetry in modern radiation treatment techniques. Since an ideal dosimeter is radiologically water equivalent, the authors investigated water equivalency and the radiological properties of three different PRESAGE formulations that differ primarily in their elemental compositions. Two of the formulations are new and have lower halogen content than the original formulation. METHODS: The radiological water equivalence was assessed by comparing the densities, interaction probabilities, and radiation dosimetry properties of the three different PRESAGE formulations to the corresponding values for water. The relative depth doses were calculated using Monte Carlo methods for 50, 100, 200, and 350 kVp and 6 MV x-ray beams. RESULTS: The mass densities of the three PRESAGE formulations varied from 5.3% higher than that of water to as much as 10% higher than that of water for the original formulation. The probability of photoelectric absorption in the three different PRESAGE formulations varied from 2.2 times greater than that of water for the new formulations to 3.5 times greater than that of water for the original formulation. The mass attenuation coefficient for the three formulations is 12%-50% higher than the value for water. These differences occur over an energy range (10-100 keV) in which the photoelectric effect is the dominant interaction. The collision mass stopping powers of the relatively lower halogen-containing PRESAGE formulations also exhibit marginally better water equivalency than the original higher halogen containing PRESAGE formulation. Furthermore, the depth dose curves for the lower halogen-containing PRESAGE formulations are slightly closer to that of water for a 6 MV beam. In the kilovoltage energy range, the depth dose curves for the lower halogen-containing PRESAGE formulations are in better agreement with water than the original PRESAGE formulation. CONCLUSIONS: Based on the results of this study, the new PRESAGE formulations with lower halogen content are more radiologically water equivalent overall than the original formulation. This indicates that the new PRESAGE formulations are better suited to clinical applications and are more accurate dosimeters and phantoms than the original PRESAGE formulation. While correction factors are still needed to convert the dose measured by the dosimeter to an absorbed dose in water in the kilovoltage energy range, these correction factors are considerably smaller for the new PRESAGE formulations compared to the original PRESAGE and the existing polymer gel dosimeters. PMID- 21626962 TI - An opposite view data replacement approach for reducing artifacts due to metallic dental objects. AB - PURPOSE: To present a conceptually new method for metal artifact reduction (MAR) that can be used on patients with multiple objects within the scan plane that are also of small sized along the longitudinal (scanning) direction, such as dental fillings. METHODS: The proposed algorithm, named opposite view replacement, achieves MAR by first detecting the projection data affected by metal objects and then replacing the affected projections by the corresponding opposite view projections, which are not affected by metal objects. The authors also applied a fading process to avoid producing any discontinuities in the boundary of the affected projection areas in the sinogram. A skull phantom with and without a variety of dental metal inserts was made to extract the performance metric of the algorithm. A head and neck case, typical of IMRT planning, was also tested. RESULTS: The reconstructed CT images based on this new replacement scheme show a significant improvement in image quality for patients with metallic dental objects compared to the MAR algorithms based on the interpolation scheme. For the phantom, the authors showed that the artifact reduction algorithm can efficiently recover the CT numbers in the area next to the metallic objects. CONCLUSIONS: The authors presented a new and efficient method for artifact reduction due to multiple small metallic objects. The obtained results from phantoms and clinical cases fully validate the proposed approach. PMID- 21626963 TI - Water equivalent path length measurement in proton radiotherapy using time resolved diode dosimetry. AB - PURPOSE: To verify water equivalent path length (WEPL) before treatment in proton radiotherapy using time resolved in vivo diode dosimetry. METHODS: Using a passively scattered range modulated proton beam, the output of a diode driving a fast current-to-voltage amplifier is recorded at a number of depths in a water tank. At each depth, a burst of overlapping single proton pulses is observed. The rms duration of the burst is computed and the resulting data set is fitted with a cubic polynomial. RESULTS: When the diode is subsequently set to an arbitrary depth and the polynomial is used as a calibration curve, the "unknown" depth is determined within 0.3 mm rms. CONCLUSIONS: A diode or a diode array, placed (for instance) in the rectum in conjunction with a rectal balloon, can potentially determine the WEPL at that point, just prior to treatment, with submillimeter accuracy, allowing the beam energy to be adjusted. The associated unwanted dose is about 0.2% of a typical single fraction treatment dose. PMID- 21626964 TI - Dose calculation for permanent prostate implants incorporating spatially anisotropic linearly time-resolving edema. AB - PURPOSE: The objectives of this study were (i) to develop a dose calculation method for permanent prostate implants that incorporates a clinically motivated model for edema and (ii) to illustrate the use of the method by calculating the preimplant dosimetry error for a reference configuration of 125I, 103Pd, and 137Cs seeds subject to edema-induced motions corresponding to a variety of model parameters. METHODS: A model for spatially anisotropic edema that resolves linearly with time was developed based on serial magnetic resonance imaging measurements made previously at our center to characterize the edema for a group of n = 40 prostate implant patients [R. S. Sloboda et al., "Time course of prostatic edema post permanent seed implant determined by magnetic resonance imaging," Brachytherapy 9, 354-361 (2010)]. Model parameters consisted of edema magnitude, delta, and period, T. The TG-43 dose calculation formalism for a point source was extended to incorporate the edema model, thus enabling calculation via numerical integration of the cumulative dose around an individual seed in the presence of edema. Using an even power piecewise-continuous polynomial representation for the radial dose function, the cumulative dose was also expressed in closed analytical form. Application of the method was illustrated by calculating the preimplant dosimetry error, RE(preplan), in a 5 x 5 x 5 cm3 volume for 125I (Oncura 6711), 103Pd (Theragenics 200), and 131Cs (IsoRay CS-1) seeds arranged in the Radiological Physics Center test case 2 configuration for a range of edema relative magnitudes (delta = [0.1, 0.2, 0.4, 0.6, 1.0]) and periods (T = [28, 56, 84] d). Results were compared to preimplant dosimetry errors calculated using a variation of the isotropic edema model developed by Chen et al. ["Dosimetric effects of edema in permanent prostate seed implants: A rigorous solution," Int. J. Radiat. Oncol., Biol., Phys. 47, 1405-1419 (2000)]. RESULTS: As expected, RE(preplan) for our edema model indicated underdosage in the calculation volume with a clear dependence on seed and calculation point positions, and increased with increasing values of delta and T. Values of RE(preplan) were generally larger near the ends of the virtual prostate in the RPC phantom compared with more central locations. For edema characteristics similar to the population average values previously measured at our center, i.e., delta = 0.2 and T = 28 d, mean values of RE(preplan) in an axial plane located 1.5 cm from the center of the seed distribution were 8.3% for 131Cs seeds, 7.5% for 103Pd seeds, and 2.2% for 125I seeds. Maximum values of RE(preplan) in the same plane were about 1.5 times greater. Note that detailed results strictly apply only for loose seed implants where the seeds are fixed in tissue and move in synchrony with that tissue. CONCLUSIONS: A dose calculation method for permanent prostate implants incorporating spatially anisotropic linearly time resolving edema was developed for which cumulative dose can be written in closed form. The method yields values for RE(preplan) that differ from those for spatially isotropic edema. The method is suitable for calculating pre- and postimplant dosimetry correction factors for clinical seed configurations when edema characteristics can be measured or estimated. PMID- 21626965 TI - Commissioning of output factors for uniform scanning proton beams. AB - PURPOSE: Current commercial treatment planning systems are not able to accurately predict output factors and calculate monitor units for proton fields. Patient specific field output factors are thus determined by either measurements or empirical modeling based on commissioning data. The objective of this study is to commission output factors for uniform scanning beams utilized at the ProCure proton therapy centers. METHODS: Using water phantoms and a plane parallel ionization chamber, the authors first measured output factors with a fixed 10 cm diameter aperture as a function of proton range and modulation width for clinically available proton beams with ranges between 4 and 31.5 cm and modulation widths between 2 and 15 cm. The authors then measured the output factor as a function of collimated field size at various calibration depths for proton beams of various ranges and modulation widths. The authors further examined the dependence of the output factor on the scanning area (i.e., uncollimated proton field), snout position, and phantom material. An empirical model was developed to calculate the output factor for patient-specific fields and the model-predicted output factors were compared to measurements. RESULTS: The output factor increased with proton range and field size, and decreased with modulation width. The scanning area and snout position have a small but non negligible effect on the output factors. The predicted output factors based on the empirical modeling agreed within 2% of measurements for all prostate treatment fields and within 3% for 98.5% of all treatment fields. CONCLUSIONS: Comprehensive measurements at a large subset of available beam conditions are needed to commission output factors for proton therapy beams. The empirical modeling agrees well with the measured output factor data. This investigation indicates that it is possible to accurately predict output factors and thus eliminate or reduce time-consuming patient-specific output measurements for proton treatments. PMID- 21626966 TI - Feminisation of the South African medical profession--not yet nirvana for gender equity. PMID- 21626968 TI - Will National Health Insurance ensure the national health? PMID- 21626967 TI - South African ischaemic stroke guideline, 2010. PMID- 21626969 TI - South African transplantation--where are we now and where should we go next? PMID- 21626970 TI - HIV prevalence in Zimbabwe dropping like a stone. PMID- 21626971 TI - SmartPhones improving clinical outcomes. PMID- 21626972 TI - Health corruption busters reveal the monster in our midst. PMID- 21626973 TI - Burns disasters--a plan for South Africa. PMID- 21626974 TI - Ethical decision making in severe paediatric burn victims. PMID- 21626975 TI - Vaginal deliveries--is there a need for documented consent? PMID- 21626976 TI - Fit for purpose? The appropriate education of health professionals in South Africa. PMID- 21626977 TI - Down syndrome in paediatric outpatient wards at Durban hospitals. PMID- 21626978 TI - Educational factors that influence the urban-rural distribution of health professionals in South Africa: a case-control study. AB - SETTING: The influence of undergraduate and postgraduate training on health professionals' career choices in favour of rural and underserved communities has not been clearly demonstrated in resource-constrained settings. OBJECTIVES: This study aimed to evaluate the influence of educational factors on the choice of rural or urban sites of practice of health professionals in South Africa. METHODS: Responses to a questionnaire on undergraduate and postgraduate educational experiences by 174 medical practitioners in rural public practice were compared with those from 142 urban public hospital doctors. Outcomes measured included specific undergraduate and postgraduate educational experiences, and non-educational factors such as family and community influences that were likely to affect the choice of the site of practice. RESULTS: Compared with urban doctors, rural respondents were significantly less experienced, more likely to be black, and felt significantly more accountable to the community that they served. They were more than twice as likely as the urban group to have been exposed to rural situations during their undergraduate training, and were also five times more likely than urban respondents to state that exposure to rural practice as an undergraduate had influenced their choice of where they practise. Urban respondents were significantly more attracted to working where they do by professional development and postgraduate education opportunities and family factors than the rural group. CONCLUSIONS: Evidence is provided that rural exposure influences the choice of practice site by health professionals in a developing country context, but the precise curricular elements that have the most effect deserve further research. PMID- 21626979 TI - The contribution of South African curricula to prepare health professionals for working in rural or under-served areas in South Africa: a peer review evaluation. AB - SETTING: The Collaboration for Health Equity through Education and Research (CHEER) was formed in 2003 to examine strategies that would increase the production of health professionals who choose to practise in rural and under served areas in South Africa. OBJECTIVES: We aimed to identify how each faculty is preparing its students for service in rural or under-served areas. METHODS: Peer reviews were conducted at all nine participating universities. A case study approach was used, with each peer review constituting its own study but following a common protocol and tools. Each research team comprised at least three reviewers from different universities, and each review was conducted over at least 3 days on site. The participating faculties were assessed on 11 themes, including faculty mission statements, resource allocation, student selection, first exposure of students to rural and under-served areas, length of exposure, practical experience, theoretical input, involvement with the community, relationship with the health service, assessment of students and research and programme evaluation. RESULTS: With a few exceptions, most themes were assessed as inadequate or adequate with respect to the preparation of students for practice in rural or under-served areas after qualification, despite implicit intentions to the contrary at certain faculties. CONCLUSIONS: Common challenges, best practices and potential solutions have been identified through this project. Greater priority must be given to supporting rural teaching sites in terms of resources and teaching capacity, in partnership with government agencies. PMID- 21626980 TI - Abdominal and pericardial ultrasound in suspected extrapulmonary or disseminated tuberculosis. AB - OBJECTIVE: Tuberculosis (TB) in patients with or without advanced HIV infection may present as smear-negative, extrapulmonary and/or disseminated forms. We studied the role of pericardial and abdominal ultrasound examinations in the determination of extrapulmonary or disseminated TB. METHODS: A prospective descriptive and analytic cross-sectional study design was used to determine the ultrasound findings of value in patients with subsequently proven TB. Ultrasound examinations were performed on 300 patients admitted to G F Jooste Hospital with suspected extrapulmonary or disseminated TB. OUTCOME MEASURES: The presence of hepatomegaly, splenomegaly, lymphadenopathy (location, size and appearance), ascites, pleural effusions, pericardial effusions and/or splenic micro-abscesses was noted. Clinical findings, microbiological and serological data were also recorded, correlated and analysed. RESULTS: Complete data sets were available for 267 patients; 91.0% were HIV positive, and 70.0% had World Health Organization clinical stage 4 disease. Active TB (determined by smear or culture) was present in 170 cases (63.7%). Ultrasonically visible abdominal lymphadenopathy over 1 cm in minimum diameter correlated with active TB in 55.3% of cases (odds ratio (OR) 2.6, 95% confidence interval (CI) 1.5 - 4.6, p = 0.0002). Ultrasonographically detected pericardial effusions (OR 2.8, 95% CI 1.6 - 5.0, p < 0.0001), ascites (OR 2.2, 95% CI 1.2 - 4.2, p = 0.005) and splenic lesions (OR 1.9, 95% CI 1.0 - 3.5, p = 0.024) also predicted active TB. CONCLUSION: Pericardial and abdominal ultrasound examinations are valuable supplementary investigations in the diagnosis of suspected extrapulmonary or disseminated TB. PMID- 21626981 TI - Postgraduate palliative care education: evaluation of a South African programme. AB - AIM: We aimed to assess the postgraduate palliative care distance education programme of the University of Cape Town (UCT) in terms of its perceived ability to influence palliative care delivery. METHODS: A mixed-methods approach, consisting of two surveys using open-ended and multiple-choice options, was conducted from January to December 2007 at the UCT School of Public Health and Family Medicine. All students registered in the programme from 2000 - 2007 were invited to participate; 83 (66.4% of all eligible participants) completed the general survey, and 41 (65.7%) of the programme's graduates completed the graduate survey. The survey scores and open-ended data were triangulated to evaluate UCT's palliative care postgraduate programme. RESULTS: General survey scores of graduates were significantly higher in 5 of the 6 categories in comparison with current students. The graduate survey indicated that curriculum and teaching strengths were in communication and dealing with challenging encounters. Graduates also stressed the need to develop a curriculum that incorporated a practical component. CONCLUSIONS: In addition to current postgraduate training, palliative care education in South Africa should be extended to undergraduate medical students, as the benefits of UCT's programme were limited to a small cohort of practitioners. PMID- 21626982 TI - Effectiveness of prenatal screening for Down syndrome on the basis of maternal age in Cape Town. AB - OBJECTIVE: The prenatal screening programme for Down syndrome (DS) in the South African public health sector remains primarily based on advanced maternal age (AMA). We assessed the changes over time and effectiveness of this screening programme within a Cape Town health district. METHODS: Retrospective analysis of the Groote Schuur Hospital Cytogenetic Laboratory and Pregnancy Counselling Clinic databases and audit of maternal delivery records at a primary health care facility. RESULTS: The number of amniocenteses performed for AMA in consecutive 5 year periods reduced progressively from 786 in 1981 - 1985 to 360 in 2001 - 2005. Comparing prenatal with neonatal diagnoses of DS, the absolute number and the proportion diagnosed prenatally have remained relatively constant over time. The Pregnancy Counselling Database showed that, of 507 women receiving genetic counselling for AMA in 2008 - 2009, 158 (3.1.1%) accepted amniocentesis--uptake has reduced considerably since the early 1990s. The audit of women delivering at a primary care facility found that only 10 (16.4%) of 61 AMA women reached genetic counselling in tertiary care: reasons included late initiation of antenatal care and low referral rates from primary care. CONCLUSION: Prenatal screening and diagnosis for DS based on AMA is working ineffectively in the Cape Town health district assessed, and this appears to be representative of a broader trend in South Africa. Inclusion of fetal ultrasound in the process of prenatal screening for DS should be explored as a way forward. PMID- 21626983 TI - Validity of oral mucosal transudate specimens for HIV testing using enzyme-linked immunosorbent assay in children in Chimanimani district, Zimbabwe. AB - OBJECTIVE: To assess the validity of oral mucosal transudate (OMT) specimens for HIV testing in children using enzyme-linked immunosorbent assay (ELISA). METHODS: A cross-sectional descriptive study was conducted as part of a community-based behavioural and HIV sero-status survey of adults and children in the Chimanimani district of Zimbabwe. Dried blood spot (DBS) and OMT samples were collected from children aged between 2 and 14 years, inclusive. Both samples were tested for HIV using the Vironostika Uniform II plus O kits. The main study outcomes were the sensitivity and specificity of OMT samples, with DBS as the gold-standard specimen. RESULTS: Paired DBS and OMT specimens were available from 1 274 (94.4%) of the 1 350 children enrolled. Using the DBS, HIV prevalence was 3.2%. Overall sensitivity of OMT was 48.8% (95% confidence interval (CI) 33.3 - 64.5), and specificity was 98.5% (95% CI 97.7 - 99.1). CONCLUSION: The overall sensitivity of OMT specimens for HIV testing in children using ELISA was low. Stratifying the analysis by sector showed that OMT samples are good specimens for HIV testing. It is important to note that factors such as the low HIV prevalence in our study population, quality of the OMT, diet and oral hygiene could have influenced the results. PMID- 21626984 TI - Ecological determinants of blindness in Nigeria: the Nigeria National Blindness and Visual Impairment Survey. AB - OBJECTIVE: To determine the prevalence and causes of visual loss in different ecological zones across Nigeria. METHODS: A population-based survey using multi stage, stratified, cluster random sampling with probability proportional to size comprising a nationally representative sample of adults aged > or = 40 years from six ecological zones. OUTCOME MEASURES: Distance vision was measured using reduced logMAR charts. Clinical examination included basic eye examination for all respondents and a detailed examination including visual fields, gonioscopy and fundus photography for those who were visually impaired or blind (i.e. presenting vision < 20/40 in the better eye). A principal cause of visual loss was assigned to all respondents with presenting vision < 20/40 in the better eye. RESULTS: A total of 15 122 persons aged > or = 40 years were enumerated, 13 599 (89.9%) of whom were examined. The prevalence of blindness varied according to ecological zone, being highest in the Sahel region (6.6%; 95% confidence interval (CI) 4.2 - 10.4) and lowest in the rain forest region (3.23%; 95% CI 2.6 - 3.9). Age/ gender-adjusted analyses showed that risk of blindness was highest in Sahel (odds ratio (OR) 3.4; 95% CI 2.1 - 5.8). More than 80% of blindness in all ecological regions was avoidable. Trachoma was a significant cause only in the Sudan savannah belt. The prevalence of all major blinding conditions was highest in the Sahel. CONCLUSIONS: The findings of this national survey may be applicable to other countries in West and Central Africa that share similar ecological zones. Onchocerciasis and trachoma are not major causes of blindness in Nigeria, possibly reflecting successful control efforts for both these neglected tropical diseases. PMID- 21626985 TI - The importance of the proceedings in thoracic endoscopy of the AIPO Study Group of Interventional Pulmonology. PMID- 21626986 TI - Organization of an endoscopic division. PMID- 21626987 TI - Bronchoscopy report: format and filing, images and exam correlation. PMID- 21626988 TI - Instruments and maintenance. PMID- 21626989 TI - Indications for diagnostic bronchoscopy in adults. PMID- 21626990 TI - Preparing of the patient, performing diagnostic flexible bronchoscopy and sampling from the airways. PMID- 21626991 TI - Transbronchial pulmonary biopsies. PMID- 21626992 TI - Bronchoalveolar lavage. Technical notes. PMID- 21626993 TI - Transbronchial needle aspiration. PMID- 21626994 TI - Autofluorescence bronchoscopy and endobronchial ultrasound. PMID- 21626995 TI - Contraindications, risks, complications in interventional pneumology. PMID- 21626996 TI - Lung transplantation and endoscopic issues. PMID- 21626997 TI - Paediatric bronchology. PMID- 21626998 TI - Bronchoscopy in intensive care unit. PMID- 21626999 TI - Palliative therapy. PMID- 21627000 TI - Bronchoscopic curative therapy. PMID- 21627001 TI - Management of post-intubation and/or tracheotomy tracheal stenoses. PMID- 21627002 TI - Airway foreign bodies. PMID- 21627003 TI - Therapeutic medical thoracoscopy. PMID- 21627004 TI - Percutaneous approach in pneumology. PMID- 21627005 TI - Advanced medical thoracoscopy. PMID- 21627006 TI - Zombie states: reconsidering the relationship between life and death instincts. AB - Where the dialectical relation of life and death instincts has become dissociated, zombie states result-in which individuals inhabit deadness as if it were a full experience of aliveness. Bypassing reservations about the speculative nature of these instincts, this paper reconsiders their relation in order to highlight certain types of clinical phenomenology that could otherwise be lost to current ways of conceptualizing aliveness and deadness. A clinical vignette illustrates particular countertransference difficulties associated with dichotomizing issues of psychic aliveness and deadness, as well as the powerful contagion associated with what the author terms zombie states. PMID- 21627007 TI - Depressive rumination in an analysand after a traumatic treatment. AB - The author attempts to understand the underpinnings of a ruminative depression that occurred in a patient after a troubled first analysis. Negative therapeutic reaction is often assumed to be the result of a patient's unconscious guilt or masochism and thus an intrapsychic phenomenon, but the author asserts that iatrogenic phenomena in the form of persistent misunderstandings and enactments that remain unanalyzed contribute to a destructive treatment experience. The analysand may relive the failed treatment again and again in his or her mind in an attempt to resolve it. The author asserts that a traumatic treatment experience can foster depressive rumination. PMID- 21627008 TI - A crisis in the analyst's life: self-containment, symbolization, and the holding space. AB - Most analysts will experience some degree of crisis in the course of their working life. This paper explores the complex interplay between the analyst's affect during a crisis in her lifeu and the affective dynamics of the patient. The central question is "who or what holds the analyst"--especially in times of crisis. Symbolization of affect, facilitated by the analyst's self-created holding environment, is seen as a vital process in order for containment to take place. In the clinical case presented, the analyst's dog was an integral part of the analyst's self-righting through this difficult period; the dog functioned as an "analytic object" within the analysis. PMID- 21627009 TI - The spit and image: a psychoanalytic dissection of a colloquial expression. AB - "He is the spit of his father" or "he is the spit and image of his father" is a colloquial expression that has graced informal English for many centuries. When a "spitting image" made an entrance in the manifest content of an analysand's dream, it became possible to add a psychoanalytic point of view to an etymological and anthropological record. After discussing both this clinical case and an "anthropological case history," the author examines the subtle but complex genesis of this colloquial expression from a speculative applied psychoanalytic perspective. PMID- 21627010 TI - Introduction to Lawrence S. Kubie's "The drive to become both sexes" (1974). PMID- 21627011 TI - The drive to become both sexes. 1974. PMID- 21627012 TI - Gender regulation. PMID- 21627013 TI - On the irreconcilable in psychic life: the role of culture in the drive to become both sexes. PMID- 21627014 TI - Japan in our hearts. PMID- 21627015 TI - The influence of CD40 ligation and interferon-gamma on functional properties of human monocyte-derived dendritic cells activated with polyinosinic-polycytidylic acid. AB - BACKGROUND/AIM: Ligation of a Toll-like receptor (TLR) by specific TLR agonists is a powerful tool for maturation induction of monocyte-derived dendritic cells (MoDCs). Studies so far have shown that the treatment of dendritic cells (DCs) with a TLR3 ligand, polyinosinic-polycytidylicacid [Poly(I:C)], may be an appropriate activation agent for obtaining mature MoDCs, competent to prime effective immune responses. However, little is known about how subsequent interaction of MoDCs with T cell-derived stimuli, such as CD40 or interferon gamma (IFN-gamma), modulates MoDC functions. Therefore, this problem was the main objective of this study. METHODS: Immature MoDCs were prepared by cultivation of monocytes from peripheral blood mononuclear cells with granulocyte macrophage colony stimulating factor (GM-CSF) and interleukin (IL)-4 for 5 days. After that, maturation was induced by the treatment of these cells with Poly(I:C) for 2 days. At day 6, immature MoDCs and Poly(I:C)-activated MoDCs were incubated either with CD40 ligand (L)-transfected J558 cells or IFN-gamma for additional 24 hours. Cytokine production was measured by ELISA and FlowCytomix Human T helper Th1/Th2 11plex. Allostimulatory capability of MoDCs was tested using an allogeneic mixed leukocyte reaction (MLR) assay. RESULTS: Immature MoDCs showed a moderate potential for stimulation of proliferation of CD4+ T cells, which was enhanced by the treatment with Poly(I:C). Ligation of CD40 or treatment with IFN-gamma of immature or Poly(I:C)-treated MoDCs significantly up-regulated their allostimulatory activity. MoDCs matured in the presence of Poly(I:C) up-regulated the production of IL-12 and IL-10, which was followed by increased levels of IFN gamma and decreased levels of IL-5 in co-cultures with allogeneic CD4+ T cells. Ligation of CD40 on immature MoDCs upregulated the production of IL-12 and IL-23 which was accompanied by increased secretion of IFN-gamma in co-culture. Stimulation of CD40 on Poly(I:C)-treated MoDCs significantly enhanced the production of IL-12, IL-23 and IL-10. However, such treated MoDCs decreased the production of IFN-gamma and IL-10 and up-regulated the secretion of IL-17. Immature MoDCs treated with IFN-gamma up-regulated IL-12, but lowered the production of IL-5 and IL-17 by CD4+ T cells. Treatment of Poly(I:C)-activated MoDCs with IFN-gamma down-regulated the production of IL-12 and up-regulated IL 10 by these cells and increased/decreased the levels of IL-10/ IFN-gamma, respectively, in co-culture with CD4+ T cells. CONCLUSION: Treatment with Poly(I:C) or ligation of CD40 on immature MoDCs induces maturation of these cells into a phenotype that supports Th1 response. Activation of CD40 on Poly(I:C) treated MoDCs shifts the immune response towards Th17. Treatment of immature MoDCs with IFN-gamma down-regulated Th2 and Th17 responses. However, addition of IFN-gamma to Poly(I:C)-activated MoDCs down-regulated Th1 response and promote T regulatory mechanisms. Each of these results may have functional and therapeutic implications. PMID- 21627016 TI - The frequency of sensitization to inhalatory allergens and concomitant rhinitis in asthmatic patients. AB - BACKGROUND/AIM: Asthma is one of the most common chronic pulmonary diseases. The number of asthmatics has been continuously increasing all over the world. Depending on its causing agent, asthma is classified as allergic and nonallergic. Asthma is often associated with other allergic diseases, and it is most commonly preceded by the symptoms of rhinitis. The aim of this study was to establish the type and frequency of allergic sensitization to inhalatory allergens, frequency of concomitant rhinitis, gender and age-related distribution of asthma, and the presence of some risk factors in patients with diagnosed asthma. METHODS: This retrospective and partially prospective analysis included 733 patients with asthma diagnosed in the Institute for Pulmonary Diseases of Vojvodina, Sremska Kamenica over the period January, 2004-December, 2008. The obtained date were statistically processed. RESULTS: Females were significantly more often affected by asthma (p < 0.05), most frequently at 20-29 years of age. A hereditary predisposition to the diseases in terms of atopy was registered in 34.9% of the examined subjects. Most patients had allergic asthma (79.5%). Sensitization to internal and external inhalatory allergens was verified in 77.5% and 67.6% of the patients respectively, and combined hypersensitivity to both allergen types in 48.8% of the patients. Rhinitis was registered in 63.9% and 28% of the patients with allergic and nonallergic asthma, respectively. Rhinitis symptoms preceded the occurrence of asthma in 60% of the patients, with the precedence of rhinitis ranging from 1 to 27 years. A high correlation between rhinitis and asthma was established for the disease of both allergic (r = 0.92) and nonallergic (r = 0.88) etiology. CONCLUSION: The majority of the patients have allergic asthma, and they are females at 20-29 years of age. Senstization to internal allergens is most common, and then to external ones. Rhinitis is the most common concomitant disease, usually preceding the occurrence of asthmatic symptoms. PMID- 21627017 TI - Significance of magnetic resonance imaging in differential diagnosis of nontraumatic brachial plexopathy. AB - BACKGROUND/AIM: Nontraumatic brachial plexopathies may be caused by primary or secondary tumors, radiation or inflammation. The aim of this study was to present the significance of MRI in revealing the cause of nontraumatic brachial plexopathy. METHODS: A two-year retrospective study included 22 patients with nontraumatic brachial plexopathy. In all the patients typical clinical findings were confirmed by upper limb neurophysiological studies. In all of them MRI of brachial plexus was performed by 1.5 T scanner in T1 and T1 FS sequence with and without contrast, as well as in T2 and T2 FS sequences. RESULTS: Seven (32%) patients had brachial plexopathy with signs of inflammatory process, 5 (23%) patients had secondary tumors, in 4 (18%) patients multifocal motor neuropathy was established and in the same number (18%) of the patients postradiation fibrosis was found. Two patients (9%) had primary neurogenic tumors. CONCLUSION: According to the results of this study MRI is a method which may determine localization and cause of brachial plexopathy. MRI can detect focal nerve lesions when other methods fail to find them. Thus, MRI has a direct impact on further diagnostic and therapeutical procedures. PMID- 21627018 TI - Optical metrology analysis of the lower jaw deformations. AB - BACKGROUND/AIM: New optical stereometric methods based on both contact and noncontact mechanisms for displacement measurement have become common methods in biomechanical behavior research of biomaterials, bone and soft tissue. The aim of this study was to register and measure possible deformations of the lower jaw (mandible) with the intact dental arch using optical metrology method. METHODS: The system for full field measurement of deformations (strains) comprised of two digital cameras for a synchronized stereoview of the specimen, and the Aramis software. RESULTS: The maximum mandibular bone strains were measured in the regions of the lower first premolar and the lower second molar. In the action force of 500 N simulated in the region of the first lower premolar the intensity of deformation was 86 microm. The value of maximum strain in the bone around the molars was 24 microm for the force of 500 N acting on the second lower molar. When it comes to premolars, 3-5 times stronger deformation was observed in the region of the first lower premolar, compared to the deformation values of the second lower premolar area. CONCLUSION: Under loading of the applied forces the measured strains were in the elastic deformation area, meanning that the dependence of force and deformity is linear. The highest values of strain measurements obtained by the optical method were found in the jaw bone tissue around the loading teeth, and the bony regions of the triangle and mental region. According to the obtained results from the Aramis processing software it can be concluded that this method is applicable in a variety of biomedical research. PMID- 21627019 TI - Selective intraarterial radionuclide therapy with Yttrium-90 (Y-90) microspheres for hepatic neuroendocrine metastases: initial experience at a single center. AB - BACKGROUND/AIM: Selective intraarterial radionuclide therapy (SIRT) with Yttrium 90 (Y-90) microspheres is also known as radioembolization and delivers high doses of radiation to hepatic tumors with minimum healthy liver exposure. The aim of this study was to present our preliminary experience in the role of liver directed radiotherapy with Y-90 microspheres for the treatment of unresectable hepatic metastases from neuroendocrine tumors (NET). METHODS: The results of SIRT in 10 patients (5 males, 5 females; mean age 48.7 years; age range 24-73 years) with metastatic liver disease from NETs during the period from April 2008 through August 2010 were reviewed. All patients had meticulous pre- and post-imaging studies as a part of their work-up procedure, as well as serologic tests of liver function to determine the extent of liver function damage. The patients who were eligible for SIRT had pretreatment visceral angiography to define and occlude non target arteries. RESULTS: The mean +/- SD administered SIR-Spheres activity was 1.49 +/- 0.42 GBq (range 0.72-2.21 GBq) in all the patients. These treatments delivered a dose of 99.73 +/- 66.36 Gy (range 49-420.8 Gy) to the target tumors. The estimated dose to the lungs and normal liver was 4.45 +/- 1.95 Gy (range 2.4 8.5 Gy) and 26.73 +/- 14.19 Gy (range 5-58.9 Gy), respectively. Overall response rate of 90% and patient tolerance was satisfactory for most patients. CONCLUSION: From our limited experience, we can conclude that SIRT with Y-90 microspheres is a safe and efficacious treatment option for patients with liver metastasis of NET without any serious side effects. PMID- 21627020 TI - Cochlear implant--speech and language development in deaf and hard of hearing children following implantation. AB - BACKGROUND/AIM: Almost 200 cochlear implantations were done in the four centers (two in Belgrade, per one in Novi Sad and Nis) in Serbia from 2002 to 2009. Less than 10% of implantees were postlingually deaf adults. The vast majority, i.e. 90% were pre- and perilingually profoundly deaf children. The aim of this study was to assess the influence of improved auditory perception due to cochlear implantation on comprehension of abstract words in children as compared with hearing impaired children with conventional hearing aids and normal hearing children. METHODS: Thirty children were enrolled in this study: 20 hearing impaired and 10 normal hearing. The vocabulary test was used. RESULTS: The overall results for the whole test (100 words) showed a significant difference in favor of the normal hearing as compared with hearing impaired children. The normal hearing children successfully described or defined 77.93% of a total of 100 words. Success rate for the cochlear implanted children was 26.87% and for the hearing impaired children with conventional hearing aids 20.32%. CONCLUSION: Testing for abstract words showed a statistically significant difference between the cochlear implanted and the hearing impaired children with hearing aids (Mann Whitney U-test, p = 0.019) implying considerable advantage of cochlear implants over hearing aids regarding successful speech development in prelingually deaf children. PMID- 21627021 TI - Metastasis of hepatocellular carcinoma presented as a tumor of the maxillary sinus and retrobulbar tumor. AB - INTRODUCTION: Hepatocellular carcinoma (HCC) is the most frequent primary malignant tumor of the liver. It is usually seen in the 6th and 7th decades of life and chronic hepatitis B is the most frequent cause. Extrahepatic metastasis of HCC is an indicator of a poor prognosis and the most common sites are lungs, bones, lymph nodes, kidneys and adrenal glands. We reported a case of isolated metastasis in the right maxilla, which had been found initially, before the tumor in the liver was diagnosed. CASE REPORT: A 70-year-old man underwent dental surgery of the upper right molar. Prolonged bleeding control was difficult for up to two weeks, so the biopsy was performed. Histopathological analysis revealed a metastatic hepatocellular carcinoma. Computerized tomography (CT) of the abdomen revealed a diffusely heterogeneous liver parenchyma with irregular borders and two foci of mass lesions. There were metastasis in the spleen and also two pathological retroperitoneal lymph nodes were detected, but no ascit, liver cirrhosis, cholestasis or portal vein thrombosis were seen. CT of the orbital and maxillary regions revealed a tumor mass in the right maxillary sinus, spreading to the alveolar sinus, nasal cavity and partially infratemporal space. A tumor mass was in the right orbit as well, infiltrating the surrounding bones and muscles. Clinically, there was proptosis of the right eye accompanied by amaurosis. The treatment started with chemotherapy based on 5-fluorouracil (sorafenib was not available). After three cycles, control CTs showed a stable disease in the liver, but progression in the right maxillary sinus and orbit. Enucleation of the right eye was performed and postoperative radiotherapy was planed. The patient deteriorated rapidly and died, about 6 months after the disease had been diagnosed. CONCLUSION: Extrahepatic metastasis of HCC represents a progressive phase of the disease with poor prognosis, so the main aim of the treatment should be palliation and care of symptoms. PMID- 21627022 TI - Bilateral Monteggia fracture in adults. AB - INTRODUCTION: In 1814 Giovanni Monteggia first described two cases of fractures of the proximal third of ulna with dislocation of the radial head. These fractures are more common in children than in adults, and mutual Monteggia fracture is a rare complication. This study presents a treatment course of a patient with bilateral Monteggia fracture. CASE REPORT: A 55-year-old patient was injured by falling in the yard. Radiography showed bilateral Monteggia fracture type II (by the Badon classification). Operative treatment of fracture was done by a compression plate on the right side and by the zuggurtung technique on the left one. Closed repositioning of the radial head was done on both sides. The patient was wearing a plaster splint for the upper arm for 21 days. After removing the fixation, the function of the elbow was determined by the Broberg Morrey score (BM) which was on the right side 45.5 and on the left side 47.5. After the proper physical therapy, four months after the surgery, BM score was 100 on the right side, and 93 on the left one. CONCLUSION: Surgical treatment and early rehabilitation is the key for the return of good function of both elbows. PMID- 21627023 TI - Preimplantation filling of tooth socket with beta-tricalcium phosphate/polylactic polyglycolic acid (beta-TCP/PLGA) root analogue: clinical and histological analysis in a patient. AB - INTRODUCTION: Bone resorption is a physiological process after tooth extraction. The use of bone substitutes to fill the tooth socket is suggested to prevent bone resorption and establish good bone architecture for implant placement. A pure beta-tricalcium phosphate coated with copolymer (polylactic-polyglycolic acid) as a root analogue, is suitable for filling tooth sockets. CASE REPORT: We presented a patient successfully treated with root analogue after extraction of the right second lower premolar. Three months later, the patient was planned for the placement of six TE ITI dental implants into the mandible. During the sugery, the biopsy of bone-like tissue from the previously treated socket was taken. All the implants were immediately loaded due to good primary stability. Histological analysis of the specimen revealed fibrous healing in the area treated with root analogue. CONCLUSION: The use of beta-tricalcium phosphate coated with copolymers after tooth extraction enables satisfactory bone architecture for consequent implant treatment. PMID- 21627024 TI - Regulation of microRNAs. Preface. PMID- 21627025 TI - MicroRNA biogenesis and function. An overview. AB - During the last decade of the 20th century a totally novel way of gene regulation was revealed. Findings that at first glance appeared freak features of plants or C. elegans turned out to be mechanistically related and deeply conserved throughout evolution. This important insight was primed by the landmark discovery of RNA interference, or RNAi, in 1998. This work started an entire novel field of research, now usually referred to as RNA silencing. The common denominator of the phenomena grouped in this field are small RNA molecules, often derived from double stranded RNA precursors, that in association with proteins of the so called Argonaute family, are capable of directing a variety of effector complexes to cognate RNA and/or DNA molecules. One of these processes is now widely known as microRNA-mediated gene silencing and I will provide a partially historical framework of the many steps that have led to our current understanding of microRNA biogenesis and function. This chapter is meant to provide a general overview of the various processes involved. For a comprehensive description of current models, I refer interested readers to the reviews and primary literature references provided in this chapter and to the further contents of this book. PMID- 21627026 TI - Regulation of pri-miRNA processing through Smads. AB - microRNAs (miRNAs) are small (-22 nucleotides (nt)), noncoding RNAs that play a critical role in diverse biological functions by modulating mRNA stability and translational control. Numerous miRNA profiling studies have indicated that the levels of miRNAs are tightly controlled during developmental stages and various pathophysiological and physiological conditions. Following transcription, the long primary miRNA transcript undergoes a series of coordinated maturation steps to generate the mature miRNA. Signaling pathways that control miRNA biogenesis and the mechanisms ofregulation, however, are not well understood. In this chapter, we will discuss the finding that signal transducers of the Transforming Growth Factor beta (TGFbeta) signaling pathway, the Smads, play a critical regulatory role in the nuclear processing of miRNAs by the RNase III-type protein Drosha. PMID- 21627027 TI - Stimulation of pri-miR-18a processing by hnRNP A1. AB - Recent evidence suggests that the canonical miRNA processing pathway can b regulated by a number of positive and negative trans-acting factors. This chapter provides an overview of hnRNP A1-mediated regulation of miR-18a biogenesis. Our laboratory has recently established that the multifunctional RNA-binding protein hnRNP A1 is required for the processing of miR-18a at the nuclear of Drosha mediated processing. By combining structural and functional analysis of RNA, we showed that hnRNP A1 regulates the processing of pri-miR-18a by binding to its terminal loop and reshaping its stem-loop structure, thus allowing for a more effective Drosha cleavage. Furthermore, we linked the event of hnRNP A1-binding to the pri-miR-18a with an unusual phylogenetic sequence conservation of its terminal loop. Bioinformatic and mutational analysis revealed that a number of pri-miRNAs have highly conserved terminal loops, which are predicted to act as landing pads for trans-acting factors influencing miRNA processing. These results underscore a previously uncharacterized role for general RNA-binding proteins as factors that facilitate the processing of specific miRNAs, revealing an additional level of complexity for the regulation of miRNA production and function. PMID- 21627028 TI - KSRP promotes the maturation of a group of miRNA precursors. AB - microRNAs (miRNAs) are small noncodingRNAs that down-regulate gene expression by reducing stability and/or translation of target mRNAs. In animals, miRNAs arise from sequential processing of hairpin primary transcripts by two RNAse III domain containing enzymes, namely Drosha and Dicer, to generate a mature form of about 22 nucleotides. In this chapter we discuss our latest findings indicating that KSRP is an integral component of both Drosha and Dicer complexes. KSRP binds to the terminal loop sequence of a subset of miRNA precursors promoting their maturation. Our data indicate that the terminal loop is a pivotal structure where activators of miRNA processing as well as repressors of miRNA processing act in a coordinated way to convert cellular signals into changes in miRNA expression processing. This uncovers a new level of complexity of miRNA mechanisms for gene expression regulation. PMID- 21627029 TI - Hormonal repression of miRNA biosynthesis through a nuclear steroid hormone receptor. AB - The maturation of primary microRNAs (pri-miRNAs) to precursor miRNAs (pre-miRNAs) is mediated by the "microprocessor" complex minimally comprimising two core components, Drosha and DGCR8. However, the roles of RNA-binding proteins associated with these core units in the large Drosha complex remain to be defined. While signal-dependent regulation of miRNA biogenesis is assumed, such regulation remains to be described. Here, we provide a short review based on our recent findings of hormonally-regulated pri-miRNA processing by nuclear estrogen receptor. PMID- 21627030 TI - Autoregulatory mechanisms controlling the Microprocessor. AB - The Microprocessor, comprising the ribonuclease Drosha and its essential cofactor, the double-stranded RNA-binding protein, DGCR8, is essential for the first step of the miRNA biogenesis pathway. It specifically cleaves double stranded RNA within stem-loop structures of primary miRNA transcripts (pri miRNAs) to generate precursor (pre-miRNA) intermediates. Pre-miRNAs are subsequently processed by Dicer to their mature 22 nt form. Thus, Microprocessor is essential for miRNA maturation, and pri-miRNA cleavage by this complex defines one end of the mature miRNA. Moreover, it is emerging that dysregulation of the Microprocessor is associated with various human diseases. It is therefore important to understand the mechanisms by which the expression of the subunits of the Microprocessor is regulated. Recent findings have uncovered a post transcriptional mechanism that maintains the integrity of the Microprocessor. These studies revealed that the Microprocessor is involved in the processing of the messenger RNA (mRNA) that encodes DGCR8. This regulatory feedback loop, along with the reported role played by DGCR8 in the stabilization of Drosha protein, is part ofa newly identified regulatory mechanism controlling Microprocessor activity. PMID- 21627031 TI - Regulation of pre-miRNA processing. AB - microRNAs are endogenously expressed 21 nucleotide noncoding RNAs. microRNA mediated regulation of the translation of specific mRNA is implicated in a range of developmental processes and pathologies. As such, miRNA expression is tightly controlled in normal development by both transcriptional and post-transcriptional mechanisms. This chapter is concerned with the control of pre-miRNA processing of individual miRNAs by specific factors. It is focussed on the regulation of a subset of miRNAs by the RNA-binding protein Lin28/LIN-28. We discuss how Lin28/LIN-28 can sequester pre-let-7 miRNA precursor to prevent Dicer-mediated processing. We describe how interaction of pre-let-7 with Lin28/ LIN-28 leads to pre-let-7 uridylation and subsequent degradation. Finally, we analyze how let-7 and Lin28/LIN-28 together act as a highly conserved developmental switch that controls stem cell differentiation in C. elegans and mammals. PMID- 21627032 TI - The effect of RNA editing and ADARs on miRNA biogenesis and function. AB - From analysis of deep-sequencing data it is apparent that sequence differences occur between the genome and miRNAs. Changes from genomic A to an apparent G in miRNA can be accounted for by the editing activity of ADARs. Questions that arise from this observation are: How many miRNAs are edited and to what frequency? Is there a specific step inthebiogenesis of miRNAs that is preferentially susceptible to editing by ADARs? However the key question is whether editing affects the downstream activity ofmiRNAs. Despite much evidence that miRNAs are edited, critical examination of the functional data shows a dearth of examples where editing has been demonstrated to actually affect the downstream miRNA activity in vivo. Even where it is demonstratedthat RNA editing can affect biogenesis or targeting of a particular miRNA, effects may be limited by redundancy within the miRNA network. PMID- 21627033 TI - MiRNA need a TRIM regulation of miRNA activity by Trim-NHL proteins. AB - Trim-NHL proteins are defined by RING, B-Box and Coiled-coil protein motifs (referred to collectively as the Trim domain) coupled to an NHL domain. The C. elegans, D. melanogaster, mouse and human Trim-NHL proteins are potential and in several cases confirmed, E3 ubiquitin ligases. Current research is focused on identifying targets and pathways for Trim-NHL-mediated ubiquitination and in assessing the contribution of the NHL protein-protein interactiondomain for function and specificity. Several Trim-NHL proteins were discovered in screens for developmental genes in model organisms; mutations in one of the family members, Trim32, cause developmental disturbances in humans. In most instances, mutations that alter protein function map to the NHL domain. The NHL domain is a scaffold for the assembly of a translational repressor complex by the Brat proto oncogene, a well-studied family member in Drosophila. The link to translational control is common to at least four Trim-NHLs that associate with miRNA pathway proteins. So far, two have been shown to repress (Mei-P26 and Lin41) and two to promote (NHL-2, Trim32) miRNA-mediated gene silencing. In this chapter we will describe structure-function relations for each of the proteins and then focus on the lessons being learned from these proteins about miRNA functions in development and in stem cell biology. PMID- 21627034 TI - Properties of the regulatory RNA-binding protein HuR and its role in controlling miRNA repression. AB - Gene expression in eukaryotes is subject to extensive regulation at posttranscriptional levels. One of the most important sites of control involves mRNA 3' untranslated regions (3'UTRs), which are recognized by RNA-binding proteins (RBPs) and microRNAs (miRNAs). These factors greatly influence translational efficiency and stability of target mRNAs and often also determine their cellular localization. HuR, a ubiquitously expressed member of the ELAV family of RBPs, has been implicated in regulation of stability and translation of over one hundred mRNAs in mammalian cells. Recent data indicate that some of the effects of HuR can be explained by its interplay with miRNAs. Binding of HuR may suppress the inhibitory effect of miRNAs interacting with the 3'UTR and redirect the repressed mRNA to polysomes for active translation. However, HuR can also synergize with miRNAs. The finding that HuR is able to disengage miRNAs from the repressed mRNA, or render them inactive, provides evidence that miRNA regulation is much more dynamic then originally anticipated. In this chapter we review properties of HuR and describe examples of the cross-talk between the protein and miRNAs, with emphasis on response of the regulation to cellular stress. PMID- 21627035 TI - Turnover of mature miRNAs and siRNAs in plants and algae. AB - microRNAs (miRNAs) and small interfering RNAs (siRNAs) play important roles in gene regulation and defense responses against transposons and viruses in eukaryotes. These small RNAs generally trigger the silencing of cognate sequences through a variety of mechanisms, including RNA degradation, translational inhibition and transcriptional repression. In the past few years, the synthesis and the mode of action of miRNAs and siRNAs have attracted great attention. However, relatively little is known about mechanisms of quality control during small RNA biogenesis as well as those that regulate mature small RNA stability. Recent studies in Arabidopsis thaliana and Caenorhabditis elegans have implicated 3'-to-5' (SDNs) and 5'-to-3' (XRN-2) exoribonucleases in mature miRNA turnover and the modulation of small RNA levels and activity. In the green alga Chlamydomonas reinhardtii, a nucleotidyltransferase (MUT68) and an exosome subunit (RRP6) are involved in the 3' untemplated uridylation and the degradation of miRNAs and siRNAs. The latter enzymes appear to function as a quality control mechanism to eliminate putative dysfunctional or damaged small RNA molecules. Several post-transcriptional modifications of miRNAs and siRNAs such as 3' terminal methylation and untemplated nucleotide additions have also been reported to affect small RNA stability. These collective findings are beginning to uncover a new layer of regulatory control in the pathways involving small RNAs. We anticipate that understanding the mechanisms of mature miRNA and siRNA turnover will have direct implications for fundamental biology as well as for applications of RNA interference technology. PMID- 21627037 TI - Epigenetic contributions in autoimmune disease. Preface. PMID- 21627036 TI - MicroRNAses and the regulated degradation of mature animal miRNAs. AB - microRNAs (miRNAs) are small noncoding RNAs that regulate numerous target mRNAs through an antisense mechanism. Initially thought to be very stable with half lives on the order of days, mature miRNAs have recently been shown to be subject to degradation by 'microRNases' (miRNases) in plants (the small RNA degrading nucleases, SDN) and animals (exoribonuclease 2/XRN-2/XRN2). Interference with these miRNA turnover pathways causes excess miRNA activity, consistent with an important contribution to miRNA homeostasis. Moreover, it is now emerging that long half-lives are not an invariant feature of miRNAs but that marked differences exist in the stabilities of individual miRNAs and that cellular states can further determine miRNA turnover rates. Although the means of regulation are still largely unclear, biochemical data suggest that target mRNA binding can stabilize miRNAs within their Argonaute (AGO) effector complexes, providing one possible mechanism that may control miRNA half-lives. We will summarize here what is known about miRNA turnover in animals and how recent discoveries have established a new dynamic of miRNA-mediated gene regulation. We will highlight some of the open questions in this emerging area of research. PMID- 21627038 TI - An introduction to epigenetics. AB - Eukaryotic genomic information is modulated by a variety ofepigenetic modifications that play both a direct role in establishing transcription profiles, modulation of DNA replication and repair processes and also indirect effects on the aforementioned processes through the organization of DNA architecture within the cell nucleus. Nowadays, the role of epigenetic modifications in regulating tissue-specific expression, genomic imprinting or X chromosome inactivation is widely recognized. In addition, the key role of epigenetic modifications during cell differentiation and development has been highlighted by the identification of a variety of epigenetic alterations in human disease. Particular attention has been focused on the study of epigenetic alterations in cancer, which is the subject of intense multidisciplinary efforts and has an impact not only in understanding the mechanisms of epigenetic regulation but also in guiding the development ofnovel therapies for cancer treatment. In addition, a number of genetic disorders such as immunodeficiency centromere instability-facial anomalies (ICF) or Rett syndromes are directly associated with defects in elements of the epigenetic machinery. More recently, epigenetic changes in cardiovascular, neurological and autoimmune disorders as well as in other genetically complex diseases have also started to emerge. All these examples illustrate the widespread association ofepigenetic alterations with disease and highlight the need of characterizing the range and extension of epigenetic changes to understand their contribution to fundamental human biological processes. PMID- 21627039 TI - Chromatin mechanisms regulating gene expression in health and disease. AB - It is now well established that the interplay of sequence-specific DNA binding proteins with chromatin components and the subsequent expression of differential genetic programs is the major determinant of developmental decisions. The last years have seen an explosion of basic research that has significantly enhanced our understanding of the basic principles of gene expression control. While many questions are still open, we are now at the stage where we can exploit this knowledge to address questions of how deregulated gene expression and aberrant chromatin programming contributes to disease processes. This chapter will give a basic introduction into the principles of epigenetics and the determinants of chromatin structure and will discuss the molecular mechanisms of aberrant gene regulation in blood cell diseases, such as inflammation and leukemia. PMID- 21627040 TI - Epigenetic control of lymphocyte differentiation. AB - Lymphocyte differentiation from haematopoietic stem cells (HSCs) is a multi-step process in which lineage fate choices are made at crucial branch points. Plasticity of common precursors is evidenced by presence of transcriptionally favourable chromatin structures at several lineage-specific loci, making them poised for further priming and regulation. Down the differentiation tree, the interplay between lineage-specific networks of transcription factors and epigenetic modifications gradually decreases the multipotency ability of precursors and increases the compromise of cells within a particular lineage. The maintenance of a cell-specific phenotype is the result of sustained gene expression programs resulting from activation of lineage-specific and repression of lineage-discrepant loci. Theperipheral functional specialisation oflymphocytes requires furtherplasticity, to allow differentiation onto short term effectors cells, orlong term memory circulating and resident cells. Impaired differentiation of lymphocytes or deregulated or unbalanced production of certain lymphocytes subsets underlies thepathogenesis of lymphoproliferation, autoimmunity and possibly immunodeficiency. Understanding epigenetic mechanisms governing lymphocyte differentiation would allow future therapeutic interventions to prevent aberrant deviations or promote beneficial cell populations. PMID- 21627041 TI - Epigenetic control in immune function. AB - This chapter describes recent advances in our understanding how epigenetic events control immune functions with emphasis on transcriptional regulation of major histocompatibility complex ClassI (MIC-I) and Class II (MHC-II) genes. MHC-I and MHC-II molecules play an essential role in the adaptive immune response by virtue of their ability to present peptides, respectively to CD8+ and CD4+ T cells. Central to the onset of an adequate immune response to pathogens is the presentation of pathogen-derived peptides in the context of MHC-II molecules by antigen presenting cells (APCs) to CD4+ T cells of the immune system. In particular dendritic cells are highly specialized APCs that are capable to activate naive T cells. Given their central role in adaptive immunity, MHC-I and MHC-II genes are regulated in a tight fashion at the transcriptional level to meet with local requirements of an effective antigen-specific immune response. In these regulatory processes the MHC2TA encoded Class II transactivator (CIITA) plays a crucial role. CIITA is essential for transcriptional activation of all MHC-I genes, whereas it plays an ancillary function in the transcriptional control of MHC-I genes. The focus of this chapter therefore will be on the transcription factors that interact with conserved cis-acting promoter elements and epigenetic mechanisms that modulate cell type-specific regulation of MHC-I, MHC-I, and MHC2TA genes. Furthermore, we will also briefly discuss how genetic and epigenetic mechanisms contribute to T helper cell differentiation. PMID- 21627042 TI - DNA methylation and B-cell autoreactivity. AB - Although not exclusive, mounting evidence supports the fact that DNA methylation at CpG dinucleotides controls B-cell development and the progressive eliminati or inactivation of autoreactive B cell. Indeed, the expression of different B ce specific factors, including Pax5, rearrangement of the B-cell receptor (BCR) and cytokine production are tightly controlled by DNA methylation. Among normal B cells, the autoreactive CD5+ B cell sub-population presents a reduced capacity to methylate its DNA that leads to the expression of normally repressed genes, such as the human endogenous retrovirus (HERV). In systemic lupus erythematosus (SLE) patients, the archetype ofautoimmune disease, autoreactive B cells are characterized by their inability to induce DNA methylation that prolongs their survival. Finally, treating B cells with demethylating drugs increased their autoreactivity. Altogether this suggests that a deeper comprehension ofDNA methylation in B cells may offer opportunities to develop new therapeutics to control autoreactive B cells. PMID- 21627043 TI - Environmental agents and autoimmune diseases. AB - Autoimmune diseases, which comprise over 80 clinically distinct conditions, are characterized by the presence of autoantibodies or autoreactive T cells directed against self structures (autoantigens). While these often incurable disorders appear to be rapidly increasing in recognition throughout the world, their rarity, heterogeneity and complex etiologies have limited our understanding of their pathogeneses. The precise mechanisms for the development of autoimmune diseases are not known, however, evidence from many complementary lines of investigation suggests that autoimmune diseases result from the interactions of both environmental and genetic risk factors. While considerable progress has been made in understanding multiple genetic risk factors for many autoimmune diseases, relatively little information is now available regarding the role of the environment in the development of these illnesses. This chapter examines the limited but growing evidence for the role of the environment in the development and progression of autoimmune diseases, the specific exposures that have been suspected of being involved, the possible mechanisms by which these agents may induce and sustain autoimmune processes and the approaches needed to better understand these issues in the future. Identifying the necessary and sufficient genetic and environmental risk factors for disease holds the promise of allowing for the prevention of some illnesses through avoidance of environmental risk factors by genetically susceptible individuals or via gene or other therapies to correct the effects of deleterious genetic risk factors in the case of unavoidable environmental agents. PMID- 21627044 TI - Epigenetic dysregulation of epstein-barr virus latency and development of autoimmune disease. AB - Epstein-Barr virus (EBV) is ahumanherpesvirus thatpersists in the memory B-cells of the majority of the world population in a latent form. Primary EBV infection is asymptomatic or causes a self-limiting disease, infectious mononucleosis. Virus latency is associated with a wide variety of neoplasms whereof some occur in immune suppressed individuals. Virus production does not occur in strict latency. The expression of latent viral oncoproteins and nontranslated RNAs is under epigenetic control via DNA methylation and histone modifications that results either in a complete silencing of the EBV genome in memory B cells, or in a cell-type dependent usage of a couple of latency promoters in tumor cells, germinal center B cells and lymphoblastoid cells (LCL, transformed by EBV in vitro). Both, latent and lytic EBV proteins elicit a strong immune response. In immune suppressed and infectious mononucleosis patients, an increased viral load can be detected in the blood. Enhanced lytic replication may result in new infection- and transformation-events and thus is a risk factor both for malignant transformation and the development of autoimmune diseases. An increased viral load or a changed presentation of a subset of lytic or latent EBV proteins that cross-react with cellular antigens may trigger pathogenic processes through molecular mimicry that result in multiple sclerosis (MS), systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). PMID- 21627045 TI - Does genomic imprinting play a role in autoimmunity? AB - In the 19th century Gregor Mendel defined the laws of genetic inheritance by crossing different types of peas. From these results arose his principle of equivalence: the gene will have the same behaviour whether it is inherited from the mother or the father. Today, several key exceptions to this principle are known, for example sex-linked traits and genes in the mitochondrial genome, whose inheritance patterns are referred to as 'non mendelian'. A third, important exception in mammals is that of genomic imprinting, where transcripts are expressed in a monoallelic fashion from only the maternal or the paternal chromosome. In this chapter, we discuss how parent-of-origin effects and genomic imprinting may play a role in autoimmunity and speculate how imprinted miRNAs may influence the expression of many target autoimmune associated genes. PMID- 21627046 TI - A new epigenetic challenge: systemic lupus erythematosus. AB - In recent years, compelling evidence has been gathered that supports a role for epigenetic alterations in the pathogenesis of systemic lupus erythematosus (SLE). Different blood cell populations of SLE patients are characterized by a global loss of DNA methylation. This process is associated with defects in ERK pathway signalling and consequent DNMT 1 downregulation. Hypomethylation of gene promoters has been described, which permits transcriptional activation and therefore functional changes in the cells and also hypomethylation of the ribosomal RNA gene cluster. Among the identified targets undergoing demethylation are genes involved in autoreactivity (ITGAL), osmotic lysis and apoptosis (PRF1, MMP14 and LCN2), antigen presentation (CSF3R), inflammation(MMP 14), B- T-cell interaction (CD70 and CD40LG) and cytokine pathways (CSF3R, IL-4, IL-6 and IFNGR2). DNA methylation inhibitors are also known to induce autoreactivity in vitro and cause a lupus-like disease in vivo. Further, altered patterns of histone modifications have been described in SLE. CD4+ lymphocytes undergo global histone H3 and H4 deacetylation and consequent skewed gene expression. Although multiple lines of evidence highlight the contribution of epigenetic alterations to the pathogenesis of lupus in genetically predisposed individuals, many questions remain to be answered. Attaining a deeper understanding of these matters will create opportunities in the promising area of epigenetic treatments. PMID- 21627047 TI - Epigenetic deregulation in rheumatoid arthritis. AB - In this chapter, we discuss the current understanding of the possible epigenetics changes that occur in rheumatoid arthritis. In particular, we describe that deregulation ofDNA methylation and histone modifications can occur in the immune system and lead to rheumatoid arthritis. In addition, we discuss the role of rheumatoid arthritis synovial fibroblasts in autoimmunity. Examples of changes in DNA methylation and histone modification occurring in synovial fibroblasts during the disease process are reviewed in this chapter. In conclusion, we discuss the possible use of epigenetic therapy and describe future experiments that can elucidate further the epigenetic changes observed in the disease. PMID- 21627048 TI - Prospects for epigenetic compounds in the treatment of autoimmune disease. AB - There is growing evidence for a role for epigenetic mechanisms in the development of autoimmune diseases. In most cases ofautoimmune disease the precise epigenetic mechanism involved remains to be resolved, however DNA hypomethylation accompanied by hypoacetylation ofhistone H3/H4 is commonly observed. Due to the reversible nature of epigenetic marks their maintenance enzymes such as DNA methyltransferases (DNMTs), histone deacetylases (HDACs) and histone lysine methyltransferases (HKMT) are attractive drug targets. Small molecule inhibitors of histone modification and DNA methylation maintenance are increasingly becoming available and will be useful chemical biological tools to dissect epigenetic mechanisms in these diseases. However, although epigenetic therapies used in cancer treatment are a promising starting point for the exploration of autoimmune disease treatment, there is a requirement for more specific and less toxic agents for these chronic diseases or for use as chemopreventative agents. PMID- 21627049 TI - Profiling epigenetic alterations in disease. AB - Nowadays, epigenetics is one of the fastest growing research areas in biomedicine. Studies have demonstrated that changes in the epigenome are not only common in cancer, but are also involved in the pathogenesis of noncancerous diseases like immunological, cardiovascular, developmental and neurological/psychiatric disorders. At the same time, during the last years, a technological revolution has taken place in the field of epigenomics, which is defined as the study of epigenetic changes throughout the whole genome. Microarray technologies and more recently, the development of next generation sequencing devices are now providing researchers with tools to draw high resolution maps of DNA methylation and histone modifications in normal tissues and diseases. This chapter will review the currently available high-throughput techniques for studying the epigenome and their applications for characterizing human diseases. PMID- 21627050 TI - Nursing students' attitudes toward video games and related new media technologies. AB - Little is known about Millennial nursing students' attitudes toward computer games and new media in nursing education and whether these attitudes differ between undergraduates and graduates. This study elicited nursing students' experience with computer games and new media, their attitudes toward various instructional styles and methods, and the role of computer games and new media technologies in nursing education. We e-mailed all nursing students enrolled in two universities to invite their participation in an anonymous cross-sectional online survey. The survey collected demographic data and participants' experience with and attitudes toward video gaming and multi-player online health care simulations. We used descriptive statistics and logistic regression to compare the differences between undergraduates and graduates. Two hundred eighteen nursing students participated. Many of the nursing students support using new media technologies in nursing education. Nurse educators should identify areas suitable for new media integration and further evaluate the effectiveness of these technologies. PMID- 21627051 TI - Teaching undergraduate students community nursing: using action research to increase engagement and learning. AB - Nurses play a pivotal role in responding to the changing needs of community health care. Therefore, nursing education must be relevant, responsive, and evidence based. We report a case study of curriculum development in a community nursing unit embedded within an undergraduate nursing degree. We used action research to develop, deliver, evaluate, and redesign the curriculum. Feedback was obtained through self-reflection, expert opinion from community stakeholders, formal student evaluation, and critical review. Changes made, especially in curriculum delivery, led to improved learner focus and more clearly linked theory and practice. The redesigned unit improved performance, measured with the university's student evaluation of feedback instrument (increased from 0.3 to 0.5 points below to 0.1 to 0.5 points above faculty mean in all domains), and was well received by teaching staff. The process confirmed that improved pedagogy can increase student engagement with content and perception of a unit as relevant to future practice. PMID- 21627052 TI - Immersion experience in undergraduate psychiatric mental health nursing. AB - Immersion practica have been used in a variety of nursing settings to help prepare culturally competent nursing students. The psychiatric setting represents a diverse culture that traditionally has not appealed to nursing students when considering career options. This article describes the experiences of baccalaureate nursing students and hospital nursing staff following a brief immersion practicum at a psychiatric facility. Seven students were assigned to one practicum for six consecutive weekdays (45 hours), without the distraction of other classes. Both the students and staff rated the experience as positive. The students believed that their ability to form therapeutic relationships, develop clinical skills, and integrate theory into clinical practice was enhanced. While nursing faculty struggle to produce more graduates, with declining clinical sites and increased competition for these sites, the brief clinical immersion may be a reasonable alternative to a traditional rotation. PMID- 21627053 TI - Evaluating the effectiveness of personal response system technology on millennial student learning. AB - As nurse educators, we must explore new technologies that capitalize on the characteristics of millennial learners. One such technology, the personal response system (PRS), is an effective way to promote active learning and increase comprehension. Few nursing studies have examined the benefits of PRS technology on student outcomes. The purpose of this study was to evaluate the effectiveness of PRS technology on learning outcomes in two sections of an undergraduate nursing research course. A crossover design compared class quiz averages between and within groups. Findings related to between and within class quiz scores were mixed, whereas the effectiveness of in-class PRS questions on paper-and-pencil quiz scores and PRS-targeted quiz items was significant. Knowledge gained from this study can be used to enhance our ability to actively engage our technologically savvy undergraduate students. By threading technology into the undergraduate curriculum, learning outcomes may be improved. PMID- 21627054 TI - Succeeding recruitment: Generation Y's next best thing. PMID- 21627055 TI - Reviewing conference abstracts. AB - This column incorporates information that provider units or organizations need to keep in mind when issuing a call for abstracts and reviewing those that have been submitted. Basic components of a successful abstract and comments about elements that will increase the likelihood of an abstract's acceptance are included. PMID- 21627056 TI - Using evidence to enhance the recovery of patients undergoing colorectal surgery: part 3. AB - This is the third part of a three-part series describing how an enhanced recovery clinical pathway uses a multidisciplinary team to reduce postoperative stress and complications, improve recovery, and decrease hospital length of stay without affecting patient safety for patients undergoing colorectal surgery. Last month, multimodal analgesia and standardized postoperative nausea and vomiting prophylaxis were discussed. This part discusses fluids, diet, tubes and drains, and early mobilization. PMID- 21627057 TI - Developing students' time management skills in clinical settings: practical considerations for busy nursing staff. AB - In clinical settings, nursing staff often find themselves responsible for students who have varying time management skills. Nurses need to respond sensitively and appropriately, and to teach nursing students how to prioritize and better allocate time. This is important not only for developing students' clinical skills but also for shaping their perceptions about the quality of the placement and their willingness to consider it as a potential work specialty. In this column, some simple, practical strategies that nurses can use to assist students with improving their time management skills are identified. PMID- 21627059 TI - A family history intervention. PMID- 21627060 TI - Occupational health nursing education. AB - Occupational health nursing academic education is generally provided at the specialty level through master's and doctoral degree preparation. This graduate education provides preparation for administrative, occupational health specialist, practitioner, academician, and researcher roles. Guided by nursing science, this specialty education encompasses a comprehensive curriculum that requires occupational health and safety coursework, practicum experience, and interdisciplinary learning. PMID- 21627061 TI - Assessment of blood pressure knowledge among workers in a midwestern manufacturing plant. A pilot study. AB - This pilot study assessed knowledge of blood pressure among employees in a midwestern manufacturing plant. The Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC 7) guidelines were used to define blood pressure categories. The Roy Adaptation Model served as the framework. Results indicated two statistically significant relationships-one between knowledge of blood pressure and employees taking blood pressure medication, and one between employees' knowledge of blood pressure readings and increased interest in learning about lifestyle modifications impacting blood pressure. Of those correctly identifying the blood pressure ranges for prehypertension, 94.4% indicated interest in learning about lifestyle modifications to improve blood pressure. Occupational health nurses in industrial settings have a unique opportunity to influence employees' blood pressure knowledge, which can propel employees to consider lifestyle modifications or wellness programs that may favorably impact indirect and direct costs. PMID- 21627063 TI - Pertussis. AB - In 2009, nearly 17,000 cases of pertussis (whooping cough) were reported in the United States, but many more go undiagnosed and unreported. Implications for occupational health nurses include assistance in early diagnosis of adult cases and prevention for employee families at home and abroad. PMID- 21627064 TI - The hydrogen-bond switch reaction of the Blrb Bluf domain of Rhodobacter sphaeroides. AB - The BlrB protein from Rhodobacter sphaeroides is a small 136 amino acid photoreceptor belonging to the BLUF family of blue light receptors. It contains merely the conserved BLUF fold responsible for binding the flavin pigment and a short C-terminal extension of unknown function. We investigated the primary photoreactions of BlrB by picosecond fluorescence and transient absorption spectroscopy. After excitation of the flavin the fluorescence decays in an H/D isotope independent manner with time constants of 21 and 390 ps, indicating a BLUF characteristic heterogeneous excited state quenched by electron transfer. By transient absorption spectroscopy, we observed a rapid relaxation of a vibrationally hot excited state within 6 ps upon excitation at 400 nm. The relaxed excited state evolves biexponentially with 18 ps (27%) and 216 ps (73%) into the signaling state spectrum indicated by a growing absorptive feature at 492 nm. Additionally, a broad triplet feature is observed as residual absorbance at a delay of 5 ns, which we attribute to derive from a significant fraction of free flavin in the sample. The photochemistry of BlrB is similar to other small BLUF proteins in respect to the fast formation of the photoproduct but does not resolve any further intermediates. We compare the photoreaction with other BLUF proteins on the basis of available spectroscopic data and crystal structures. An arginine close to the C2?O carbonyl of the flavin is likely to be a key determinant for the fast electron transfer in BlrB. Additionally, the orientation of the electron-donating tyrosine in respect to the flavin might play a role in the so far unique kinetic separation of the semiquinonic intermediates in Slr1694. PMID- 21627066 TI - Concise total synthesis of calothrixins A and B. AB - The concise total synthesis of calothrixins A and B has been accomplished by utilizing the one-pot formation of hexatriene as a key intermediate via the palladium-catalyzed tandem cyclization/cross-coupling reaction of triethyl(indol 2-yl)borate. In another key transformation, the indolo[3,2-j]phenanthridine core was prepared in high yield via Cu(I)-mediated 6pi-electrocyclization. PMID- 21627067 TI - Molecules for charge-based information storage. AB - The inexorable drive to miniaturize information storage and processing devices has fueled the dreams of scientists pursuing molecular electronics: researchers in the field envisage exquisitely tailored molecular materials fulfilling the functions now carried out by semiconductors. A bottom-up assembly of such all molecular devices would complement, if not supplant, the present top-down lithographic procedures of modern semiconductor fabrication. Short of these grand aspirations, a more near-term objective is to construct hybrid architectures wherein molecules are incorporated in semiconductor-based devices. Such a combined approach exploits the advantages of molecules for selected device functions while retaining the well-developed lithographic approaches for fabrication of the overall chip. In this Account, we survey more than a decade of results from our research programs to employ porphyrin molecules as charge storage elements in hybrid semiconductor-molecular dynamic random access memory. Porphyrins are attractive for a variety of reasons: they meet the stability criteria for use in real-world applications, they are readily prepared and tailored synthetically, they undergo read-write processes at low potential, and they store charge for extended periods (up to minutes) in the absence of applied potential. Porphyrins typically exhibit two cationic redox states. Molecular architectures with greater than two cationic redox states are achieved by combinations of porphyrins in a variety of structures (for example, dyads, wherein the porphyrins have distinct potentials, triple deckers, and dyads of triple deckers). The incorporation of porphyrins in hybrid architectures has also required diverse tethers (alkyl, alkenyl, alkynyl, aryl, and combinations thereof) and attachment groups (alcohol, thiol, selenol, phosphonate, and hydrocarbon) for linkage to a variety of surfaces (Au, Si, SiO(2), TiN, Ge, and so forth). The porphyrins as monolayers exhibit high charge density and are robust to high-temperature excursions (400 degrees C for 30 min) under inert atmosphere conditions. Even higher charge densities, which are invaluable for device applications, were achieved by in situ formation of porphyrin polymers or by stepwise growth of porphyrin-imide oligomers. The various molecular architectures have been investigated by diverse surface characterization methods, including ellipsometry, atomic force microscopy, FTIR spectroscopy, and X-ray photoelectron spectroscopy, as well as a variety of electrochemical methods. These studies have further revealed that the porphyrin layers are robust under conditions of deposition of a top metal contact. The results to date indicate the superior features of selected molecular architectures for molecular electronics applications. The near-term utilization of such materials depends on further work for appropriate integration in semiconductor-based devices, whereas ultimate adoption may depend on advances that remain far afield, such as the development of fully bottom-up assembly processes. PMID- 21627068 TI - Copper-catalyzed arylation of 1H-perfluoroalkanes. AB - A general method has been developed for arylation of readily available 1H perfluoroalkanes. The method employs aryl iodide and 1H-perfluoroalkane reagents, DMPU solvent, TMP(2)Zn base, and a copper chloride/phenanthroline catalyst. Preliminary mechanistic studies are reported. PMID- 21627069 TI - Seasonal variation of defense-related gene expression in leaves from Bois noir affected and recovered grapevines. AB - Although Bois noir is one of the main phytoplasma diseases of grapevine, the gene expression and enzyme activities that underlie physiological changes occurring in symptomatic and recovered (with spontaneous or induced symptom remission) plants are mostly unknown. Bois noir symptomatic leaves (September 2006, 2007) and symptomless leaves from infected symptomatic plants (September 2007) of Sangiovese (moderately susceptible) and Chardonnay (highly susceptible) cultivars were collected. Moreover, leaves from infected symptomless plants of both cultivars were harvested in June 2007. Leaves from recovered plants were also collected in the same periods. In recovered plants of both cultivars, class III chitinase and almost every time phenylalanine ammonia-lyase and chalcone synthase expression were increased for all collection periods. In symptomatic leaves of both cultivars, the expressions of the same genes were up-regulated and also those of beta-1,3-glucanase and flavanone 3-hydroxylase. The activities of chitinase, phenylalanine ammonia-lyase, beta-1,3-glucanase, and superoxide dismutase generally correlated with gene expression. For the moderately susceptible Sangiovese, the defense genes were generally up-regulated in both symptomatic and symptomless leaves (for all collection periods). This behavior was not observed in the highly susceptible Chardonnay, in which changes in gene expression were linked to evident symptom display. Therefore, the physiological response of the plants to this pathogen infection appear to be the reason for the resistance of the cultivar to the disease. PMID- 21627070 TI - Advancing next-generation proteomics through computational research. PMID- 21627072 TI - Chemical bonding in cesium uranyl chloride based on the experimental electron density distribution. AB - Details of the electron density distribution in Cs(2)UO(2)Cl(4) have been obtained from an accurate X-ray diffraction experiment at 20 K. The electron density was described with the Hansen-Coppens multipole model. Topological analysis of the electron density confirms that the U-O bond is probably a triple bond, the U-Cl bonds are incipient covalent interactions, and the Cs-Cl and Cs-O interactions are of the closed-shell type. The results obtained serve as a proof of principle that electron density features related to chemical bonding may be obtained from X-ray data for even the heaviest elements. PMID- 21627073 TI - Perchlorate in soybean sprouts (Glycine max L. Merr.), water dropwort (Oenanthe stolonifera DC.), and lotus (Nelumbo nucifera Gaertn.) root in South Korea. AB - The occurrence of perchlorate in soybean sprouts (Glycine max L. Merr), water dropwort (Oenanthe stolonifera DC.), and lotus (Nelumbo nucifera Gaertn.) root, which are commonly consumed by people in South Korea, was determined by using an ion chromatograph coupled with a tandem mass spectrometer. For soybean sprouts (11 samples), perchlorate was detected in most (91%) of the samples at various concentrations of up to 78.4 MUg/kg dry weight (DW); the mean concentration was 35.2 MUg/kg DW. For water dropwort, of the 13 samples examined, four showed concentrations that were above the limit of quantification (LOQ). The mean perchlorate concentration was 20.7 MUg/kg DW, and the highest perchlorate value was 39.9 MUg/kg DW. Of the six lotus root samples examined, only one exhibited a detectable perchlorate concentration (17.3 MUg/kg DW). For the accumulation experiments with artificially contaminated solutions, the concentrations of perchlorate in soybean sprouts gradually increased with the increase of perchlorate concentration in the solution. However, there was a decrease in the bioconcentration factor as the perchlorate concentration in the solution increased. PMID- 21627074 TI - Traceable multifunctional micellar nanocarriers for cancer-targeted co-delivery of MDR-1 siRNA and doxorubicin. AB - In this article we report on the development of polymeric micelles that can integrate multiple functions in one system, including the capability to accommodate a combination of therapeutic entities with different physicochemical properties (i.e., siRNA and doxorubicin; DOX), passive and active cancer targeting, cell membrane translocation, and pH-triggered drug release. A micellar system was constructed from degradable poly(ethylene oxide)-block-poly(epsilon caprolactone) (PEO-b-PCL) block copolymers with functional groups on both blocks. The functional group on the PCL block was used to incorporate short polyamines for complexation with siRNA or to chemically conjugate DOX via a pH-sensitive hydrazone linkage. A virus mimetic shell was conferred by attaching two ligands, i.e., the integrin alphavbeta3-specific ligand (RGD4C) for active cancer targeting and the cell-penetrating peptide TAT for membrane activity. This system was used to improve the efficacy of DOX in multidrug-resistant MDA-MB-435 human tumor models that overexpress P-glycoprotein (P-gp), by simultaneous intracellular delivery of DOX and siRNA against P-gp expression. The carrier was tagged with near-infrared fluorescent imaging probes to provide a means to follow the fate of the system in vivo upon intravenous administration. Dy677-labeled siRNA was also used to assess the in vivo stability of the siRNA carrier. This multifunctional polymeric micellar system was shown to be capable of DOX and siRNA delivery to their intracellular targets, leading to the inhibition of P-gp mediated DOX resistance in vitro and targeting of alphavbeta3-positive tumors in vivo. PMID- 21627075 TI - Highly luminescent mixed-metal Pt(II)/Ir(III) complexes: bis-cyclometalation of 4,6-diphenylpyrimidine as a versatile route to rigid multimetallic assemblies. AB - The proligand 4,6-di-(4-tert-butylphenyl)pyrimidine LH(2) can undergo cycloplatination with K(2)PtCl(4) at one of the two aryl rings to give, after treatment with sodium acetylacetonate, a mononuclear complex Pt(N^C-LH)(acac) (denoted Pt). If an excess of K(2)PtCl(4) is used, a dinuclear complex of the form [Pt(acac)](2){MU-(N^C-L-N^C)} (Pt(2)) is obtained instead, where the pyrimidine ring acts as a bridging unit. Alternatively, the mononuclear complex can undergo cyclometalation with a different metal ion. Thus, reaction of Pt with IrCl(3).3H(2)O (2:1 ratio) leads, after treatment with sodium acetylacetonate, to an unprecedented mixed-metal complex of the form Ir{MU-(N^C-L N^C)Pt(acac)}(2)(acac) (Pt(2)Ir). The mononuclear iridium complex Ir(N^C LH)(2)(acac) (Ir) has also been prepared for comparison. The UV-visible absorption and photoluminesence properties of the four complexes and of the proligand have been investigated. The complexes are all highly luminescent, with quantum yields of around 0.5 in solution at room temperature. The introduction of the additional metal centers is found to lead to a substantial red-shift in absorption and emission, with lambda(max) in the order Pt < Pt(2) < Ir < Pt(2)Ir. The trend is interpreted with the aid of electrochemical data and density functional theory calculations, which suggest that the red-shift is due primarily to a progressive stabilization of the lowest unoccupied molecular orbital (LUMO). The radiative decay constant is also increased. This versatile design strategy may offer a new approach for tuning and optimizing the luminescence properties of d-block metal complexes for contemporary applications. PMID- 21627076 TI - Polyphenols from evening primrose ( Oenothera paradoxa ) defatted seeds induce apoptosis in human colon cancer Caco-2 cells. AB - Polyphenols extracted from evening primrose seeds (industrial waste product) were studied as apoptosis inducers in human colorectal adenocarcinoma Caco-2 and HT-29 cell lines and in rat normal intestinal IEC-6 cells. The extract dose-dependently inhibited the growth of Caco-2, HT-29, and IEC-6 cells. However, nuclear DNA fragmentation characteristic of apoptosis was observed only in Caco-2. After 72 h of incubation with the extract at 150 MUM gallic acid equivalents (44.1 MUg extract/mL), Caco-2 cell numbers decreased to 19% of control and 48.8% of the cells were identified by flow cytometry as apoptotic. Under the same conditions only 8% of HT-29 cells and 12.6% of IEC-6 cells exhibited hypodiploid DNA content. The effects of the extract and its fractions on phosphatidylserine exposure and cell membrane integrity were assessed by high content screening image cytometry. The fractions strongly and dose-dependently reduced Caco-2 cell numbers, whereas HT-29 and IEC-6 cells were affected to lesser extents. PMID- 21627077 TI - A highly reusable rhodium catalyst-organic framework for the intramolecular cycloisomerization of 1,6-enynes. AB - The intramolecular cycloisomerization of 1,6-enynes in 95-99% ee is reported using an immobilized Rh catalyst-organic framework synthesized from alternating ring-opening metathesis polymerization (altROMP) assembly. The framework was reused up to seven times, and it was used in high turnover number (TON) batch reactions. The catalyst provided the highest TONs to date (up to 890) for the cycloisomerizations, with catalyst loadings ranging from 0.2 to 0.06 mol %. PMID- 21627078 TI - Metabolomics profiling of cell culture media leading to the identification of riboflavin photosensitized degradation of tryptophan causing slow growth in cell culture. AB - As more protein biopharmaceuticals are produced using mammalian cell culture techniques, it becomes increasingly important for the biopharmaceutical industry to have tools to characterize the cell culture media and evaluate its impact on the cell culture performance. Exposure of the cell culture media to light, temperature stress, or adventitious introduction of low-level organisms during preparation can lead to the generation of chemical degradants or metabolites of the media components, which are potentially detrimental to the cell culture process. In this work, we applied a liquid chromatography-mass spectrometry based metabolomics methodology for the investigation of a media lot used for a mammalian cell culture process that had resulted in low growth rate and failure to meet required viable cell density (VCD). The study led to the observation of increased levels of tryptophan oxidation products and a riboflavin degradant, lumichrome, in the malfunctioning media lot, relative to working media lots. A compound found 7-fold higher in the working media lots appeared to be tetrahydropentoxyline, a condensation product of glucose and tryptophan. A second compound found at an over 50-fold higher level in the malfunctioning media lot with a proposed molecular formula of C(21)H(17)N(3)O(3) from high-resolution mass spectrometry (HRMS) analysis remains unknown, although it is confirmed to be a degradant of tryptophan in the media. A study of the cell culture media performed under stress conditions using fluorescent light and heat showed that the media powder was highly resistant to light-induced degradation, while solution media could be easily degraded after brief light exposure. It is therefore suspected that inadvertent exposure of the media to light during preparation and storage has resulted in the poor performance of the media causing the low growth and VCD in the cell culture process. PMID- 21627079 TI - Controlling nanoparticle formation via sizable cages of supramolecular soft materials. AB - We present a new generic strategy to fabricate nanoparticles in the "cages" within the fibrous networks of supramolecular soft materials. As the cages can be acquired by a design-and-production manner, the size of nanoparticles synthesized within the cages can be tuned accordingly. To implement this idea, both selenium and silver were chosen for the detailed investigation. It follows that the sizes of selenium and silver nanoparticles can be controlled by tuning the pore size of the fiber networks in the material. When the concentration of the gelator is high enough, monodisperse nanoparticles can be prepared. More interestingly, the morphology of the nanoparticles can be altered: silver disks can be formed when the concentrations of both the gelator and silver nitrate are sufficiently low. As the fiber network serves as a physical barrier and semisolid support for the nanoparticles, the stability in the aqueous media and the ease of application of these nanoparticles can be substantially enhanced. This robust surfactant-free approach will not only allow the controlled fabrication of nanoparticles, but also can be applied to the fabrication of composite materials for robust applications. PMID- 21627080 TI - Increased cross-platform microarray data set correlation via substrate independent nanofilms. AB - Recently developed substrate-independent nanofilms were utilized to generate highly concordant cross-platform microarray data sets on a diverse set of materials such as glass, mica, silicon, and polymer. Using model DNA and protein dose-response assays, the number of cross-platform data sets exhibiting high correlation (>0.98) increased from 33% to 86% when utilizing platforms coated with substrate-independent nanofilms as opposed to traditional surface coatings such as aminosilane and poly-L-lysine. Furthermore, it is shown how the surface properties of the substrate-independent nanofilms can be tailored through secondary modifications to improve assay performance, demonstrating a viable approach for standardized and rapid biochip development through common, yet upgradable, cross-platform interfaces. PMID- 21627081 TI - Synthetic protein targeting by the intrinsic biorecognition functionality of poly(ethylene glycol) using PEG antibodies as biohybrid molecular adaptors. AB - Biointerfaces capable of biological recognition and specificity are sought after for conferring bioinspired functionality onto synthetic biomaterials systems. This is important for biosensing, bioseparations, and biomedical materials. Here, we demonstrate how intrinsic polymer-protein interactions between highly localized polyethylene glycol (PEG) brushes and PEG-binding antibodies can be used for sorting specific biomolecules from complex bulk biological fluids to synthetic nanoscale targets. A principal feature lies with the antifouling property of PEG that prevents unspecific binding. Exclusive access is provided by anti-PEG, which acts as a biohybrid molecular adaptor that sifts out and targets specific IgG "cargo" from solution to the PEG. The PEG can be reversibly washed and targeted in blood serum, which suggests potential benefits in technological applications. Moreover, anti-PEG binding triggers a stimuli-responsive conformational collapse in the PEG brush, thereby imparting an intrinsic "smart" biorecognition functionality to the PEG that can considerably impact its use as an antifouling biomaterial. PMID- 21627082 TI - EPR detection of an electron scavenging contaminant in irradiated deoxyoligonucleotides: one-electron reduced benzoyl. AB - Our lab investigated damage of DNA due to ionizing radiation using electron paramagnetic resonance (EPR). Through studies focused on one-electron-reduction of synthetic oligodeoxynucleotides containing only thymine and adenine, we discovered the significant presence of a contaminant in all samples. The contaminant was observed to have a reduction potential greater than that of thymine. In addition, the contaminant yielded a sharp EPR singlet when it was one electron reduced that interfered with the distinctive doublet of one-electron reduced thymine. We determined that the contaminant contained a benzoyl group, a chemical used in to protect the amine group of adenine during oligodeoxynucleotide synthesis. Derivatives of benzoyl and 16 different oligomer sequences were prepared in a LiCl glass and studied using EPR after X-irradiating at 4K. This treatment selectively creates one-electron reduced radicals. Synthetic derivatives were used to develop an EPR benchmark of the benzoyl radical. Using this, along with the known spectra of one-electron reduced nucleobases, we performed component analysis of the EPR signal from each sample. This analysis revealed that 2-9% of adenines, in the commercially synthesized oligomers delivered to us, were left contaminated with benzoyl. We concluded that the presence of benzoyl is a potential source of error in a variety of experiments utilizing synthesized oligodeoxynucleotides. PMID- 21627083 TI - Polyphenol composition of a functional fermented tea obtained by tea-rolling processing of green tea and loquat leaves. AB - Phenolic constituents of a new functional fermented tea produced by tea-rolling processing of a mixture (9:1) of tea leaves and loquat leaves were examined in detail. The similarity of the phenolic composition to that of black tea was indicated by high-performance liquid chromatography comparison with other tea products. Twenty-five compounds, including three new catechin oxidation products, were isolated, and the structures of the new compounds were determined to be (2R) 2-hydroxy-3-(2,4,6-trihydroxyphenyl)-1-(3,4,5-trihydroxyphenyl)-1-propanone 2-O gallate, dehydrotheasinensin H, and acetonyl theacitrin A by spectroscopic methods. In addition, theacitrinin A and theasinensin H were obtained for the first time from commercial tea products. Isolation of these new and known compounds confirms that reactions previously demonstrated by in vitro model experiments actually occur when fresh tea leaves are mechanically distorted and bruised during the production process. PMID- 21627084 TI - Microscope measurements for the transient formation of W/O emulsions of sodium bis(2-ethylhexyl) sulfosuccinate in the dodecane/water interfacial region. AB - The present study investigated the transient formation of water-in-oil (W/O) emulsions of sodium bis(2-ethylhexyl) sulfosuccinate (aerosol OT, AOT) in a dodecane/water interfacial region and the anomalous uptake of water in the dodecane phase by in situ bright-field optical microscopy and water concentration measurements in detail. The hydrodynamic radius of the individual W/O emulsions in the dodecane phase was determined to be 0.1-1.2 MUm from the analysis of their diffusion behavior; they are much larger than common W/O microemulsions (a few nanometers in radius). At first, they were formed spontaneously in the dodecane/water interfacial region without shaking, and they diffused away into the dodecane phase. Then, almost all of them vanished at the interface by fusion. Their number and the water concentration in the dodecane phase increased first and then decreased gradually. The formation mechanism was discussed with estimated concentration profiles of AOT and water molecules, which suggests that larger W/O emulsions of 0.01-0.44 MUm in radius can be formed in the dodecane phase near the interface (within 2 MUm) because the concentration of AOT becomes lower than that of water there. PMID- 21627085 TI - One-step selective chemistry for silicon-on-insulator sensor geometries. AB - A one-step functionalization process has been developed for oxide-free channels of field effect transistor structures, enabling a self-selective grafting of receptor molecules on the device active area, while protecting the nonactive part from nonspecific attachment of target molecules. Characterization of the self organized chemical process is performed on both Si(100) and SiO(2) surfaces by infrared and X-ray photoelectron spectroscopy, atomic force microscopy, and electrical measurements. This selective functionalization leads to structures with better chemical stability, reproducibility, and reliability than current SiO(2)-based devices using silane molecules. PMID- 21627086 TI - Protein denaturation detected by time-of-flight secondary ion mass spectrometry. AB - In the present work we investigate the denaturation of a functional protein, horseradish peroxidase (HRP), under various experimental conditions using time-of flight secondary ion mass spectrometry. HRP was immobilized on TiO(2), and the samples were stored under different conditions. The activity of the enzyme was assessed colorimetrically and compared to ToF-SIMS spectra. We show that denaturation of the protein can be monitored using the ToF-SIMS signal of the disulfide bonds, which is related to the tertiary structure of the protein. As disulfide bonds appear in a vast range of proteins, the present findings may be of wide significance; i.e., a tool is provided that can allow the investigation of the presence of an active protein structure by a comparably simple surface analytical method. PMID- 21627087 TI - Inhibitory effect of a glycoprotein isolated from golden oyster mushroom ( Pleurotus citrinopileatus ) on the lipopolysaccharide-induced inflammatory reaction in RAW 264.7 macrophage. AB - Mushrooms have become an important source of natural antitumor, antiviral, antibacterial, immunomodulatory, and anti-inflammatory agents. Golden oyster mushroom, Pleurotus citrinopileatus , is a common mushroom in oriental countries for human consumption. The present study investigated the anti-inflammatory reaction of the bioactive nonlectin glycoprotein (PCP-3A) isolated from the fresh fruiting body of this mushroom. Western blot analysis on LPS-induced iNOS, COX-2, and NF-kappaB expressions in RAW 264.7 cells as affected by PCP3-A was performed to elucidate the mechanism of NO and PGE2 reduction. The results showed that PCP 3A failed to affect RAW 264.7 viability at a concentration up to 6.25 MUg/mL, but inhibited LPS (1 MUg/mL)-induced expression, and that PCP-3A inhibited the production of NO and PGE2 in LPS-activated macrophages via the down-regulation of certain pro-inflammatory mediators, including iNOS and NF-kappaB. PMID- 21627088 TI - Letter to the editor regarding the article by Paganelli et al. PMID- 21627089 TI - Application of circular dichroism spectroscopy in the study of mixed-valence asymmetric ruthenium polypyridyl complexes. AB - Circular dichroism (CD) spectroelectrochemistry is used to determine the extent of singly occupied molecular orbital delocalization in mixed-valence multinuclear complexes, specifically the mixed-valence Ru(II)Ru(III) states of the four diastereoisomers of [(Ru(bpy)(2))(2)(bpt)](3+) [1; bpy = 2,2'-bipyridyl and bpt = 3,5-bis(pyrid-2'-yl)-1,2,4-triazolato anion]. The complex was found to be stable to thermal racemization in the three oxidation states, but photoracemization in the Ru(II)Ru(II) state was observed. PMID- 21627090 TI - Dioxygen and hydrogen peroxide reduction with hemocyanin model complexes. AB - Three copper polypyridyl complexes were examined as electrocatalysts for the oxygen reduction reaction (ORR): a Cu-N(3) complex, [Cu-[tris(6-methylpyridin-2 yl)methane]-(NCMe)]PF(6) (1); a related Cu(2)N(6) derivative, [Cu(2)-[1,2-bis(6 (bis(6-methylpyridin-2-yl)methyl)pyridin-2-yl)ethane]-(NCMe)(2)](PF(6))(2) (2); and the CuN(4) species [Cu-[tris(pyridin-2-ylmethyl)amine]](ClO(4))(2) [3](ClO(4))(2). Compared to other copper complexes, [3](ClO(4))(2) exhibits the highest reported ORR onset potential for a Cu complex of 0.53 V vs reversible hydrogen electrode at pH 1. The Cu(2)N(6) hemocyanin model is more active than the CuN(3), but both are less active than the CuN(4) complex. The results indicate that copper polypyridyl complexes are promising cathode catalysts for ORR. PMID- 21627091 TI - Probing multivalent host-guest interactions between modified polymer layers by direct force measurement. AB - The adhesion behavior between modified polysaccharide layers capable of forming host-guest complexes has been determined by direct force measurements with the atomic force microscope (AFM). Polysaccharides bearing either host or guest moieties were obtained by derivatization of chitosan with pendant beta cyclodextrin (CD) and adamantane (AD) moieties, respectively. These modified polysaccharides were covalently immobilized either to flat surfaces or to AFM probes. The number of interacting polymer segments has been reduced significantly by covalently immobilizing chitosan to an AFM-tip with small radius and measuring the forces between the protruding polymer segments and a chitosan layer immobilized to a flat surface. By this approach, it was possible to determine the interaction between polymer layers on the level of single polymer strands. To separate contributions to the adhesion due to the formation of host-guest complexes from unspecific interactions, we performed measurements between various combinations of chitosan derivatives. With the same polymer probe of adamantane modified chitosan, the interaction against a number of different chitosan layers has been determined, including ones that are not able to form host-guest complexes, such as unmodified chitosan or beta-cyclodextrin modified chitosan, which has been blocked previously by addition of adamantane. The resulting adhesion behavior has been analyzed in terms of the total work of adhesion, the number of rupture events, and the corresponding lengths of the polymer segments as well as rupture forces. A clear difference has been found for systems where the formation of host-guest complexes is possible in comparison to the absence of specific multivalent interaction between the polysaccharide layers. In particular, the work of adhesion is increasing up to an order of magnitude upon the formation of host-guest complexes between the chitosan layers. PMID- 21627092 TI - Chiral Bronsted acid-catalyzed asymmetric trisubstituted aziridine synthesis using alpha-diazoacyl oxazolidinones. AB - Despite the remarkable advances in catalytic asymmetric aziridinations over the past decades, establishing a general procedure for the stereoselective synthesis of trisubstituted aziridines has remained an elusive goal. Chiral N-triflyl phosphoramide-catalyzed reactions of N-alpha-diazoacyl oxazolidinones and N-Boc imines were developed as a solution to this unmet challenge. PMID- 21627093 TI - A density functional theory and quantum theory of atoms-in-molecules analysis of the stability of Ni(II) complexes of some amino alcohol ligands. AB - The structure of the complexes of the type [Ni(L)(H(2)O)(2)](2+), where L is an amino alcohol ligand, L = N,N'-bis(2-hydroxyethyl)-ethane-1,2-diamine (BHEEN), N,N'-bis(2-hydroxycyclohexyl)-ethane-1,2-diamine (Cy(2)EN), and N,N'-bis(2 hydroxycyclopentyl)-ethane-1,2-diamine, (Cyp(2)EN) were investigated at the X3LYP/6-31+G(d,p) level of theory both in the gas phase and in solvent (CPCM model) to gain insight into factors that control the experimental log K(1) values. We find that (i) analyses based on Bader's quantum theory of atoms in molecules (QTAIM) are useful in providing significant insight into the nature of metal-ligand bonding and in clarifying the nature of weak "nonbonded" interactions in these complexes and (ii) the conventional explanation of complex stability in these sorts of complexes (based on considerations of bond lengths, bite angles and H-clashes) could be inadequate and indeed might be misleading. The strength of metal-ligand bonds follows the order Ni-N > Ni-OH >= Ni-OH(2); the bonds are predominantly ionic with some covalent character decreasing in the order Ni-N > Ni-OH > Ni-OH(2), with Ni-OH(2) being close to purely ionic. We predict that the cis complexes are preferred over the trans complexes because of (i) stronger bonding to the alcoholic O-donor atoms and (ii) more favorable intramolecular interactions, which appear to be important in determining the conformation of a metal-ligand complex. We show that (i) the flexibility of the ligand, which controls the Ni-OH bond length, and (ii) the ability of the ligand to donate electron density to the metal are likely to be important factors in determining values of log K(1). We find that the electron density at the ring critical point of the cyclopentyl moieties in Cyp(2)EN is much higher than that in the cyclohexyl moieties of Cy(2)EN and interpret this to mean that Cyp(2)EN is a poorer donor of electron density to a Lewis acid than Cy(2)EN. PMID- 21627094 TI - Far-infrared absorption of PbSe nanorods. AB - Measurements of the far-infrared absorption spectra of PbSe nanocrystals and nanorods are presented. As the aspect ratio of the nanorods increases, the Frohlich sphere resonance splits into two peaks. We analyze this splitting with a classical electrostatic model, which is based on the dielectric function of bulk PbSe but without any free-carrier contribution. Good agreement between the measured and calculated spectra indicates that resonances in the local field factors underlie the measured spectra. PMID- 21627095 TI - Using nitrogen isotope fractionation to assess the oxidation of substituted anilines by manganese oxide. AB - We explored the N isotope fractionation associated with the oxidation of substituted primary aromatic amines, which are often the position of initial attack in transformation processes of environmental contaminants. Apparent (15)N kinetic isotope effects, AKIE(N), were determined for the oxidation of various substituted anilines in suspensions of manganese oxide (MnO(2)) and compared to reference experiments in homogeneous solutions and at electrode surfaces, as well as to density functional theory calculations of intrinsic KIE(N)for electron and hydrogen atom transfer reactions. Owing to the partial aromatic imine formation after one-electron oxidation and corresponding increase in C-N bond strength, AKIE(N)-values were inverse, substituent-dependent, and confined to the range between 0.992 and 0.999 in agreement with theory. However, AKIE(N)-values became normal once the fraction of cationic species prevailed owing to (15)N-equilibrium isotope effects, EIE(N), of 1.02 associated with N atom deprotonation. The observable AKIE(N)-values are substantially modulated by the acid/base pre equilibria of the substituted anilines and isotope fractionation may even vanish under conditions where normal EIE(N) and inverse AKIE(N) cancel each other out. The pH-dependent trends of the AKIE(N)-values provide a new line of evidence for the identification of contaminant degradation processes via oxidation of primary aromatic amino groups. PMID- 21627096 TI - Intrinsic response time of graphene photodetectors. AB - Graphene-based photodetectors are promising new devices for high-speed optoelectronic applications. However, despite recent efforts it is not clear what determines the ultimate speed limit of these devices. Here, we present measurements of the intrinsic response time of metal-graphene-metal photodetectors with monolayer graphene using an optical correlation technique with ultrashort laser pulses. We obtain a response time of 2.1 ps that is mainly given by the short lifetime of the photogenerated carriers. This time translates into a bandwidth of ~262 GHz. Moreover, we investigate the dependence of the response time on gate voltage and illumination laser power. PMID- 21627097 TI - Ultrasensitive ambient mass spectrometric analysis with a pin-to-capillary flowing atmospheric-pressure afterglow source. AB - The advent of ambient desorption/ionization mass spectrometry has resulted in a strong interest in ionization sources that are capable of direct analyte sampling and ionization. One source that has enjoyed increasing interest is the flowing atmospheric-pressure afterglow (FAPA). The FAPA has been proven capable of directly desorbing/ionizing samples in any phase (solid, liquid, or gas) and with impressive limits of detection (<100 fmol). The FAPA was also shown to be less affected by competitive-ionization matrix effects than other plasma-based sources. However, the original FAPA design exhibited substantial background levels, cluttered background spectra in the negative-ion mode, and significant oxidation of aromatic analytes, which ultimately compromised analyte identification and quantification. In the present study, a change in the FAPA configuration from a pin-to-plate to a pin-to-capillary geometry was found to vastly improve performance. Background signals in positive- and negative ionization modes were reduced by 89% and 99%, respectively. Additionally, the capillary anode strongly reduced the amount of atomic oxygen that could cause oxidation of analytes. Temperatures of the gas stream that interacts with the sample, which heavily influences desorption capabilities, were compared between the two sources by means of IR thermography. The performance of the new FAPA configuration is evaluated through the determination of a variety of compounds in positive- and negative-ion mode, including agrochemicals and explosives. A detection limit of 4 amol was found for the direct determination of the agrochemical ametryn and appears to be spectrometer-limited. The ability to quickly screen for analytes in bulk liquid samples with the pin-to-capillary FAPA is also shown. PMID- 21627098 TI - Fast monitoring of species-specific peptide biomarkers using high-intensity focused-ultrasound-assisted tryptic digestion and selected MS/MS ion monitoring. AB - A new strategy for the fast monitoring of peptide biomarkers is described. It is based on the use of accelerated in-solution trypsin digestions under an ultrasonic field provided by high-intensity focused ultrasound (HIFU) and the monitoring of several peptides by selected MS/MS ion monitoring in a linear ion trap mass spectrometer. The performance of the method was established for the unequivocal identification of all commercial fish species belonging to the Merlucciidae family. Using a particular combination of only 11 peptides, resulting from the HIFU-assisted tryptic digestion of the thermostable proteins parvalbumins, the workflow allowed the unequivocal identification of these closely related fish species in any seafood product, including processed and precooked products, in less than 2 h. The present strategy constitutes the fastest method for peptide biomarker monitoring. Its application for food quality control provides to the authorities an effective and rapid method of food authentication and traceability to guarantee the quality and safety to the consumers. PMID- 21627099 TI - X-ray nanodiffraction on a single SiGe quantum dot inside a functioning field effect transistor. AB - For advanced electronic, optoelectronic, or mechanical nanoscale devices a detailed understanding of their structural properties and in particular the strain state within their active region is of utmost importance. We demonstrate that X-ray nanodiffraction represents an excellent tool to investigate the internal structure of such devices in a nondestructive way by using a focused synchotron X-ray beam with a diameter of 400 nm. We show results on the strain fields in and around a single SiGe island, which serves as stressor for the Si channel in a fully functioning Si-metal-oxide semiconductor field-effect transistor. PMID- 21627100 TI - Defect engineering in cubic cerium oxide nanostructures for catalytic oxidation. AB - Traditional nanostructured design of cerium oxide catalysts typically focuses on their shape, size, and elemental composition. We report a different approach to enhance the catalytic activity of cerium oxide nanostructures through engineering high density of oxygen vacancy defects in these catalysts without dopants. The defect engineering was accomplished by a low pressure thermal activation process that exploits the nanosize effect of decreased oxygen storage capacity in nanostructured cerium oxides. PMID- 21627101 TI - Multiple-wavelength focusing of surface plasmons with a nonperiodic nanoslit coupler. AB - A novel type of multiple-wavelength focusing plasmonic coupler based on a nonperiodic nanoslit array is designed and experimentally demonstrated. An array of nanoslits patterned on a thin metal film is used to couple free-space light into surface plasmon polaritons (SPPs) and simultaneously focus different wavelength SPPs into arbitrary predefined locations in the two-dimensional plane. We design and fabricate a compact triplexer on a glass substrate with an integrated silicon photodetector. The photocurrent spectra demonstrate that the incident light is effectively coupled to SPPs and routed into three different focal spots depending on the wavelength. The proposed scheme provides a simple method of building wavelength-division multiplexing and spectral filtering elements, integrated with other plasmonic and optoelectronic devices. PMID- 21627102 TI - Label-free, single protein detection on a near-infrared fluorescent single-walled carbon nanotube/protein microarray fabricated by cell-free synthesis. AB - Excessive sample volumes continue to be a major limitation in the analysis of protein-protein interactions, motivating the search for label-free detection methods of greater sensitivity. Herein, we report the first chemical approach for selective protein recognition using fluorescent single-walled carbon nanotubes (SWNTs) enabling label-free microarrays capable of single protein detection. Hexahistidine-tagged capture proteins directly expressed by cell-free synthesis on SWNT/chitosan microarray are bound to a Ni(2+) chelated by Nalpha,Nalpha bis(carboxymethyl)-L-lysine grafted to chitosan surrounding the SWNT. The Ni(2+) acts as a proximity quencher with the Ni(2+)/SWNT distance altered upon docking of analyte proteins. This ability to discern single protein binding events decreases the apparent detection limit from 100 nM, for the ensemble average, to 10 pM for an observation time of 600 s. This first use of cell-free synthesis to functionalize a nanosensor extends this method to a virtually infinite number of capture proteins. To demonstrate this, the SWNT microarrays are used to analyze a network of 1156 protein-protein interactions in the staurosporine-induced apoptosis of SH-SY5Y cells, confirming literature predictions. PMID- 21627103 TI - Reversible acetylation and inactivation of Mycobacterium tuberculosis acetyl-CoA synthetase is dependent on cAMP. AB - Recent proteomics studies have revealed that protein acetylation is an abundant and evolutionarily conserved post-translational modification from prokaryotes to eukaryotes. Although an astonishing number of acetylated proteins have been identified in those studies, the acetyltransferases that target these proteins remain largely unknown. Here we characterized MSMEG_5458, one of the GCN5-related N-acetyltransferases (GNAT's) in Mycobacterium smegmatis, and show that it is a protein acetyltransferase (MsPat) that specifically acetylates the epsilon-amino group of a highly conserved lysine residue in acetyl-CoA synthetase (ACS) with a k(cat)/K(m) of nearly 10(4) M(-1) s(-1). This acetylation results in the inactivation of ACS activity. Lysine acetylation by MsPat is dependent on 3',5' cyclic adenosine monophosphate (cAMP), an important second messenger, indicating that MsPat is a downstream target of the intracellular cAMP signaling pathway. To the best of our knowledge, this is the first protein acetyltransferase in mycobacteria that both is dependent on cAMP and targets a central metabolic enzyme by a specific post-translational modification. Since cAMP is synthesized by adenylate cyclases (AC's) that sense various environmental signals, we hypothesize that the acetylation and inactivation of ACS is important for mycobacteria to adjust to environmental changes. In addition, we show that Rv1151c, a sirtuin-like deacetylase in Mycobacterium tuberculosis, reactivates acetylated ACS through an NAD(+)-dependent deacetylation. Therefore, Pat and the sirtuin-like deacetylase in mycobacteria constitute a reversible acetylation system that regulates the activity of ACS. PMID- 21627104 TI - Precision and accuracy in the quantitative analysis of biological samples by accelerator mass spectrometry: application in microdose absolute bioavailability studies. AB - Determination of the pharmacokinetics and absolute bioavailability of an experimental compound, SCH 900518, following a 89.7 nCi (100 MUg) intravenous (iv) dose of (14)C-SCH 900518 2 h post 200 mg oral administration of nonradiolabeled SCH 900518 to six healthy male subjects has been described. The plasma concentration of SCH 900518 was measured using a validated LC-MS/MS system, and accelerator mass spectrometry (AMS) was used for quantitative plasma (14)C-SCH 900518 concentration determination. Calibration standards and quality controls were included for every batch of sample analysis by AMS to ensure acceptable quality of the assay. Plasma (14)C-SCH 900518 concentrations were derived from the regression function established from the calibration standards, rather than directly from isotopic ratios from AMS measurement. The precision and accuracy of quality controls and calibration standards met the requirements of bioanalytical guidance (U.S. Department of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research, Center for Veterinary Medicine. Guidance for Industry: Bioanalytical Method Validation (ucm070107), May 2001. http://www.fda.gov/downloads/Drugs/GuidanceCompilanceRegulatoryInformation/Guidan es/ucm070107.pdf ). The AMS measurement had a linear response range from 0.0159 to 9.07 dpm/mL for plasma (14)C-SCH 900158 concentrations. The CV and accuracy were 3.4-8.5% and 94-108% (82-119% for the lower limit of quantitation (LLOQ)), respectively, with a correlation coefficient of 0.9998. The absolute bioavailability was calculated from the dose-normalized area under the curve of iv and oral doses after the plasma concentrations were plotted vs the sampling time post oral dose. The mean absolute bioavailability of SCH 900518 was 40.8% (range 16.8-60.6%). The typical accuracy and standard deviation in AMS quantitative analysis of drugs from human plasma samples have been reported for the first time, and the impact of these parameters on quantitative analysis was further assessed using the Z factor. The use of the lowest achievable LLOQ(Z=0) derived from statistical analysis of the control and low-concentration standards for AMS measurements is proposed in future studies. PMID- 21627105 TI - Crystal structures of aspartate aminotransferase reconstituted with 1 deazapyridoxal 5'-phosphate: internal aldimine and stable L-aspartate external aldimine. AB - The 1.8 A resolution crystal structures of Escherichia coli aspartate aminotransferase reconstituted with 1-deazapyridoxal 5'-phosphate (deazaPLP; 2 formyl-3-hydroxy-4-methylbenzyl phosphate) in the internal aldimine and L aspartate external aldimine forms are reported. The L-aspartate.deazaPLP external aldimine is extraordinarily stable (half-life of >20 days), allowing crystals of this intermediate to be grown by cocrystallization with L-aspartate. This structure is compared to that of the alpha-methyl-L-aspartate.PLP external aldimine. Overlays with the corresponding pyridoxal 5'-phosphate (PLP) aldimines show very similar orientations of deazaPLP with respect to PLP. The lack of a hydrogen bond between Asp222 and deazaPLP, which serves to "anchor" PLP in the active site, releases strain in the deazaPLP internal aldimine that is enforced in the PLP internal aldimine [Hayashi, H., Mizuguchi, H., Miyahara, I., Islam, M. M., Ikushiro, H., Nakajima, Y., Hirotsu, K., and Kagamiyama, H. (2003) Biochim. Biophys. Acta1647, 103] as evidenced by the planarity of the pyridine ring and the Schiff base linkage with Lys258. Additionally, loss of this anchor causes a 10 degrees greater tilt of deazaPLP toward the substrate in the external aldimine. An important mechanistic difference between the L-aspartate.deazaPLP and alpha-methyl-L-aspartate.PLP external aldimines is a hydrogen bond between Gly38 and Lys258 in the former, positioning the catalytic base above and approximately equidistant between Calpha and C4'. In contrast, in the alpha methyl-L-aspartate.PLP external aldimine, the epsilon-amino group of Lys258 is rotated ~70 degrees to form a hydrogen bond to Tyr70 because of the steric bulk of the methyl group. PMID- 21627106 TI - Comparative analysis of predictive models for nongenotoxic hepatocarcinogenicity using both toxicogenomics and quantitative structure-activity relationships. AB - The primary testing strategy to identify nongenotoxic carcinogens largely relies on the 2-year rodent bioassay, which is time-consuming and labor-intensive. There is an increasing effort to develop alternative approaches to prioritize the chemicals for, supplement, or even replace the cancer bioassay. In silico approaches based on quantitative structure-activity relationships (QSAR) are rapid and inexpensive and thus have been investigated for such purposes. A slightly more expensive approach based on short-term animal studies with toxicogenomics (TGx) represents another attractive option for this application. Thus, the primary questions are how much better predictive performance using short-term TGx models can be achieved compared to that of QSAR models, and what length of exposure is sufficient for high quality prediction based on TGx. In this study, we developed predictive models for rodent liver carcinogenicity using gene expression data generated from short-term animal models at different time points and QSAR. The study was focused on the prediction of nongenotoxic carcinogenicity since the genotoxic chemicals can be inexpensively removed from further development using various in vitro assays individually or in combination. We identified 62 chemicals whose hepatocarcinogenic potential was available from the National Center for Toxicological Research liver cancer database (NCTRlcdb). The gene expression profiles of liver tissue obtained from rats treated with these chemicals at different time points (1 day, 3 days, and 5 days) are available from the Gene Expression Omnibus (GEO) database. Both TGx and QSAR models were developed on the basis of the same set of chemicals using the same modeling approach, a nearest-centroid method with a minimum redundancy and maximum relevancy-based feature selection with performance assessed using compound-based 5-fold cross-validation. We found that the TGx models outperformed QSAR in every aspect of modeling. For example, the TGx models' predictive accuracy (0.77, 0.77, and 0.82 for the 1-day, 3-day, and 5-day models, respectively) was much higher for an independent validation set than that of a QSAR model (0.55). Permutation tests confirmed the statistical significance of the model's prediction performance. The study concluded that a short-term 5-day TGx animal model holds the potential to predict nongenotoxic hepatocarcinogenicity. PMID- 21627107 TI - Correcting for methane interferences on delta2H and delta18O measurements in pore water using H2Oliquid-H2Ovapor equilibration laser spectroscopy. AB - Cavity ring-down spectroscopy (CRDS) is a new and evolving technology that shows great promise for isotopic delta(18)O and delta(2)H analyses of pore water from equilibrated headspace H(2)O vapor from environmental and geologic cores. We show that naturally occurring levels of CH(4) can seriously interfere with CRDS spectra, leading to erroneous delta(18)O and delta(2)H results for water. We created a new CRDS correction algorithm to account for CH(4) concentrations typically observed in subsurface and anaerobic environments, such as ground waters or lake bottom sediments. We subsequently applied the correction method to a series of geologic cores that contain CH(4). The correction overcomes the spectral interference and provides accurate pore water delta(18)O and delta(2)H values with acceptable precision levels as well as accurate concentrations of CH(4). PMID- 21627109 TI - Bioactive neolignans and lignans from the bark of Machilus robusta. AB - Sixteen new neolignans and lignans (1-16), together with 12 known analogues, have been isolated from an ethanol extract of the bark of Machilus robusta. Compounds 1 and 2 showed activity against HIV-1 replication in vitro, with IC(50) values of 2.52 and 2.01 MUM, respectively. At 10 MUM, 6, 8, and 9 reduced dl-galactosamine induced hepatocyte (WB-F344 cells) damage, and 9 could additionally attenuate rotenone-induced PC12 cell damage. The known compounds (-)-pinoresinol (17) and (+)-lyoniresinol (18) were active against serum deprivation induced PC12 cell damage. PMID- 21627108 TI - Identification of ostruthin from Peucedanum ostruthium rhizomes as an inhibitor of vascular smooth muscle cell proliferation. AB - Inhibition of vascular smooth muscle cell (VSMC) proliferation is of substantial interest in combating cardiovascular disease. A dichloromethane extract from the rhizomes of Peucedanum ostruthium, a traditionally used Austrian medicinal plant with anti-inflammatory properties, was examined for a putative antiproliferative activity in rat aortic VSMC. This extract inhibited serum (10%)-induced VSMC proliferation concentration dependently. Further identification and biological testing of its major constituents revealed that the coumarin ostruthin (7) is the major antiproliferative substance. In summary, a new bioactivity of P. ostruthium rhizomes is described, and 7 has been identified as the responsible compound. PMID- 21627110 TI - Stabilization of the second oxyanion intermediate by 1,4-dihydroxy-2-naphthoyl coenzyme A synthase of the menaquinone pathway: spectroscopic evidence of the involvement of a conserved aspartic acid. AB - 1,4-Dihydroxy-2-naphthoyl-coenzyme A (DHNA-CoA) synthase, or MenB, catalyzes an intramolecular Claisen condensation involving two oxyanion intermediates in the biosynthetic pathway of menaquinone, an essential respiration electron transporter in many microorganisms. Here we report the finding that the DHNA-CoA product and its analogues bind and inhibit the synthase from Escherichia coli with significant ultraviolet--visible spectral changes, which are similar to the changes induced by deprotonation of the free inhibitors in a basic solution. Dissection of the structure--affinity relationships of the inhibitors identifies the hydroxyl groups at positions 1 (C1-OH) and 4 (C4-OH) of DHNA-CoA or their equivalents as the dominant and minor sites, respectively, for the enzyme--ligand interaction that polarizes or deprotonates the bound ligands to cause the observed spectral changes. In the meantime, spectroscopic studies with active site mutants indicate that C4-OH of the enzyme-bound DHNA-CoA interacts with conserved polar residues Arg-91, Tyr-97, and Tyr-258 likely through a hydrogen bonding network that also includes Ser-161. In addition, site-directed mutation of the conserved Asp-163 to alanine causes a complete loss of the ligand binding ability of the protein, suggesting that the Asp-163 side chain is most likely hydrogen-bonded to C1-OH of DHNA-CoA to provide the dominant polarizing effect. Moreover, this mutation also completely eliminates the enzyme activity, strongly supporting the possibility that the Asp-163 side chain provides a strong stabilizing hydrogen bond to the tetrahedral oxyanion, which takes a position similar to that of C1-OH of the enzyme-bound DHNA-CoA and is the second high energy intermediate in the intracellular Claisen condensation reaction. Interestingly, both Arg-91 and Tyr-97 are located in a disordered loop forming part of the active site of all available DHNA-CoA synthase structures. Their involvement in the interaction with the small molecule ligands suggests that the disordered loop is folded in interaction with the substrates or reaction intermediates, supporting an induced-fit catalytic mechanism for the enzyme. PMID- 21627111 TI - A free-energy landscape for coupled folding and binding of an intrinsically disordered protein in explicit solvent from detailed all-atom computations. AB - The N-terminal repressor domain of neural restrictive silencer factor (NRSF) is an intrinsically disordered protein (IDP) that binds to the paired amphipathic helix (PAH) domain of mSin3. An NMR experiment revealed that the minimal binding unit of NRSF is a 15-residue segment that adopts a helical structure upon binding to a cleft of mSin3. We computed a free-energy landscape of this system by an enhanced conformational sampling method, all-atom multicanonical molecular dynamics. The simulation started from a configuration where the NRSF segment was fully disordered and distant from mSin3 in explicit solvent. In the absence of mSin3, the disordered NRSF segment thermally fluctuated between hairpins, helices, and bent structures. In the presence of mSin3, the segment bound to mSin3 by adopting the structures involved in the isolated state, and non-native and native complexes were formed. The free-energy landscape comprised three superclusters, and free-energy barriers separated the superclusters. The native complex was located at the center of the lowest free-energy cluster. When NRSF landed in the largest supercluster, the generated non-native complex moved on the landscape to fold into the native complex, by increasing the interfacial hydrophobic contacts and the helix content. When NRSF landed in other superclusters, the non-native complex overcame the free-energy barriers between the various segment orientations in the binding cleft of mSin3. Both population shift and induced-fit (or induced-folding) mechanisms work cooperatively in the coupled folding and binding. The diverse structural adaptability of NRSF may be related to the hub properties of the IDP. PMID- 21627113 TI - Photoinduced reaction of hydrogen-end-capped polyynes with iodine molecules. AB - Hydrogen-end-capped polyynes, H(C=C)(n)H (n = 5-7), were photoirradiated in the presence of iodine molecules in nonpolar solvents to find a dramatic change in the UV/vis absorption spectrum. Absorption bands of polyynes in the UV and the band of I(2) in the visible region disappeared, whereas weaker bands of polyynes were intensified in the near UV region. The emerging features are associated with vibronic bands in the symmetry-forbidden transition of the linear polyyne molecule. Stoichiometry for the reaction, i.e., C(12)H(2) + nI(2) -> C(12)H(2)I(2n), was determined to be n = 3 from the concentration-dependence experiment. (13)C NMR spectra for 1:3 mixture of polyyne and iodine molecules, namely C(10)H(2)/3I(2) and C(12)H(2)/3I(2), exhibited five and six lines, respectively, all shifted to lower fields compared to those in the case without iodine. After removing excess I(2) by reductive reagent of Na(2)SO(3), recovery of the missing absorption bands for the components of C(10)H(2) and I(2) was observed. These observations strongly support the formation of an unique molecular complex of C(2n)H(2)I(6) (n = 5-7) upon photoabsorption by an I(2) molecule within a cluster of polyyne and iodine molecules, C(2n)H(2)(I(2))(m) (m >= 3). PMID- 21627112 TI - Biomedical applications of tetrazine cycloadditions. AB - Disease mechanisms are increasingly being resolved at the molecular level. Biomedical success at this scale creates synthetic opportunities for combining specifically designed orthogonal reactions in applications such as imaging, diagnostics, and therapy. For practical reasons, it would be helpful if bioorthogonal coupling reactions proceeded with extremely rapid kinetics (k > 10(3) M(-1) s(-1)) and high specificity. Improving kinetics would minimize both the time and amount of labeling agent required to maintain high coupling yields. In this Account, we discuss our recent efforts to design extremely rapid bioorthogonal coupling reactions between tetrazines and strained alkenes. These selective reactions were first used to covalently couple conjugated tetrazine near-infrared-emitting fluorophores to dienophile-modifed extracellular proteins on living cancer cells. Confocal fluorescence microscopy demonstrated efficient and selective labeling, and control experiments showed minimal background fluorescence. Multistep techniques were optimized to work with nanomolar concentrations of labeling agent over a time scale of minutes: the result was successful real-time imaging of covalent modification. We subsequently discovered fluorogenic probes that increase in fluorescence intensity after the chemical reaction, leading to an improved signal-to-background ratio. Fluorogenic probes were used for intracellular imaging of dienophiles. We further developed strategies to react and image chemotherapeutics, such as trans-cyclooctene taxol analogues, inside living cells. Because the coupling partners are small molecules (<300 Da), they offer unique steric advantages in multistep amplification. We also describe recent success in using tetrazine reactions to label biomarkers on cells with magneto-fluorescent nanoparticles. Two-step protocols that use bioorthogonal chemistry can significantly amplify signals over both one-step labeling procedures as well as two-step procedures that use more sterically hindered biotin-avidin interactions. Nanoparticles can be detected with fluorescence or magnetic resonance techniques. These strategies are now being routinely used on clinical samples for biomarker profiling to predict malignancy and patient outcome. Finally, we discuss recent results with tetrazine reactions used for in vivo molecular imaging applications. Rapid tetrazine cycloadditions allow modular labeling of small molecules with the most commonly used positron emission tomography isotope, (18)F. Additionally, recent work has applied this reaction directly in vivo for the pretargeted imaging of solid tumors. Future work with tetrazine cycloadditions will undoubtedly lead to optimized protocols, improved probes, and additional biomedical applications. PMID- 21627114 TI - Density functional theory study of the mechanism and origins of stereoselectivity in the asymmetric Simmons-Smith cyclopropanation with Charette chiral dioxaborolane ligand. AB - Asymmetric Simmons-Smith reaction using Charette chiral dioxaborolane ligand is a widely applied method for the construction of enantiomerically enriched cyclopropanes. The detailed mechanism and the origins of stereoselectivity of this important reaction were investigated using density functional theory (DFT) calculations. Our computational studies suggest that, in the traditional Simmons Smith reaction conditions, the monomeric iodomethylzinc allyloxide generated in situ from the allylic alcohol and the zinc reagent has a strong tendency to form a dimer or a tetramer. The tetramer can easily undergo an intramolecular cyclopropanation to give the racemic cyclopropane product. However, when a stoichiometric amount of Charette chiral dioxaborolane ligand is employed, monomeric iodomethylzinc allyloxide is converted into an energetically more stable four-coordinated chiral zinc/ligand complex. The chiral complex has the zinc bonded to the CH(2)I group and coordinated by three oxygen atoms (one from the allylic alcohol and the other two oxygen atoms from the carbonyl oxygen and the ether oxygen in the dioxaborolane ligand), and it can undergo the cyclopropanation reaction easily. Three key factors influencing the enantioselectivity have been identified through examining the cyclopropanation transition states: (1) the torsional strain along the forming C-C bond, (2) the 1,3-allylic strain caused by the chain conformation, and (3) the ring strain generated in the transition states. In addition, the origin of the high anti diastereoselectivity for the substituent on the zinc reagent and the hydroxymethyl group of the allylic alcohol has been rationalized through analyzing the steric repulsion and the ring strain in the cyclopropanation transition states. PMID- 21627115 TI - Latticelike smectic liquid crystal phase in a rigid-rod helical polyisocyanide with mesogenic pendants. AB - We report a unique macromolecule consisting of a rodlike helical polyisocyanide backbone with a narrow molecular weight distribution and rigid mesogenic chiral pendants linked via a flexible spacer that exhibits lyotropic nematic and latticelike new smectic (lat-Sm) liquid crystal phases at different concentrations. The unprecedented lat-Sm phase is associated with the smectic ordering of both the stiff polymer backbone and the rigid-rod side groups. A detailed investigation of the films using X-ray scattering and atomic force microscopy revealed a novel tilted smectic layer structure of the polymer backbone aligned perpendicular to the smectic layer of the mesogenic pendants, which arrange in an antiparallel overlapping interdigitated manner. PMID- 21627117 TI - Au36(SPh)23 nanomolecules. AB - A new core size protected completely by an aromatic thiol, Au(36)(SPh)(23), is synthesized and characterized by MALDI-TOF mass spectrometry and UV-visible spectroscopy. The synthesis involving core size changes is studied by MS, and the complete ligand coverage by aromatic thiol group is shown by NMR. PMID- 21627116 TI - Semiquinone-iron complex of photosystem II: EPR signals assigned to the low-field edge of the ground state doublet of QA*-Fe2+ and QB*-Fe2+. AB - The quinone-iron complex of the electron acceptor complex of Photosystem II was studied by EPR spectroscopy in Thermosynechococcus elongatus. New g ~ 2 features belonging to the EPR signal of the semiquinone forms of the primary and secondary quinone, i.e., Q(A)(*-)Fe(2+) and Q(B)(*-)Fe(2+), respectively, are reported. In previous studies, these signals were missed because they were obscured by the EPR signal arising from the stable tyrosyl radical, TyrD(*). When the TyrD(*) signal was removed, either by chemical reduction or by the use of a mutant lacking TyrD, the new signals dominated the spectrum. For Q(A)(*-)Fe(2+), the signal was formed by illumination at 77 K or by sodium dithionite reduction in the dark. For Q(B)(* )Fe(2+), the signal showed the characteristic period-of-two variations in its intensity when generated by a series of laser flashes. The new features showed relaxation characteristics comparable to those of the well-known features of the semiquinone-iron complexes and showed a temperature dependence consistent with an assignment to the low-field edge of the ground state doublet of the spin system. Spectral simulations are consistent with this assignment and with the current model of the spin system. The signal was also present in Q(B)(*-)Fe(2+) in plant Photosystem II, but in plants, the signal was not detected in the Q(A)(*-)Fe(2+) state. PMID- 21627119 TI - 3-haloquinolines by Friedlander reaction of alpha-haloketones. AB - A general approach to 3-fluoro-, 3-chloro-, and 3-bromoquinolines which relies on organosilane-promoted Friedlander reaction of alpha-haloketones is described. The scope of the methylene component as well as influence of the organosilane component on the outcome of the reaction is studied. The method can be used under parallel synthesis conditions. PMID- 21627118 TI - Functional virus-based polymer-protein nanoparticles by atom transfer radical polymerization. AB - Viruses and virus-like particles (VLPs) are useful tools in biomedical research. Their defined structural attributes make them attractive platforms for engineered interactions over large molecular surface areas. In this report, we describe the use of VLPs as multivalent macroinitiators for atom transfer radical polymerization. The introduction of chemically reactive monomers during polymerization provides a robust platform for post-synthetic modification via the copper-catalyzed azide-alkyne cycloaddition reaction. These results provide the basis to construct nanoparticle delivery vehicles and imaging agents using protein-polymer conjugates. PMID- 21627120 TI - Synthesis and antimalarial activity of 2-guanidino-4-oxoimidazoline derivatives. AB - A series of 2-guanidino-4-oxoimidazoline (deoxo-IZ) derivatives was prepared and showed potent antimalarial activities in rodent and Rhesus models. Compound 8e, the most potent analogues of this series, is the first non-8-aminoqinoline antimalarial that demonstrated radical curative activity in non-human primate by oral route and showed causal prophylactic activity comparable to that of the commonly used clinical drugs in Rhesus monkeys infected with sporozoites of Plasmodium cynomolgi. The metabolic stability and metabolites profile indicated that the new deoxo-IZ derivatives (8) may act as prodrugs of the corresponding IZ (1 and 2) derivatives. PMID- 21627121 TI - Irreversible Nek2 kinase inhibitors with cellular activity. AB - A structure-based approach was used to design irreversible, cysteine-targeted inhibitors of the human centrosomal kinase, Nek2. Potent inhibition of Nek2 kinase activity in biochemical and cell-based assays required a noncatalytic cysteine residue (Cys22), located near the glycine-rich loop in a subset of human kinases. Elaboration of an oxindole scaffold led to our most selective compound, oxindole propynamide 16 (JH295). Propynamide 16 irreversibly inhibited cellular Nek2 without affecting the mitotic kinases, Cdk1, Aurora B, or Plk1. Moreover, 16 did not perturb bipolar spindle assembly or the spindle assembly checkpoint. To our knowledge, 16 is the first small molecule shown to inactivate Nek2 kinase activity in cells. PMID- 21627122 TI - Photoinduced cyclization of 3-acyl-2-halo-1-[(omega-phenylethynyl)alkyl]indoles to azaheterocyclo[1,2,3-lm]-fused benzo[c]carbazoles. AB - A one-pot synthesis of azaheterocyclo[1,2,3-lm]-fused benzo[c]carbazoles (2 and 3) has been developed by photocyclization of 3-acyl-2-halo-1-[(omega phenylethynyl)alkyl] indoles (1) in good to excellent yields. All products are formed from 1 via two sequential photocyclization reactions. Two products, 9 chloro-7,8-dihydro-6H-benzo[c]pyrido[1,2,3-lm]carbazole (2a-h) and 7,8-dihydro-6H benzo[c]pyrido[1,2,3-lm]carbazole (3a-h), are produced in the photocyclization of 2-halo-1-[(omega-phenylethynyl)alkyl]indole-3-carbaldehydes (1a-h). In contrast, only products 2a-h are produced in the photocyclization of 3-acetyl-2-chloro-1 [(omega-phenylethynyl)alkyl]indole-3-carbaldehydes (1o-t). The 9-H in 3a-h (n = 2) does originate from the formyl group in 1a-h via 1,5-hydrogen shift. The structures of three new products, 9-bromo-7,8-dihydro-6H-benzo[c]pyrido[1,2,3 lm]carbazole (2b), 9-chloro-10-methyl-7,8-dihydro-6H-benzo[c]pyrido[1,2,3 lm]carbazole (3h) and 12-chloro-7,8-dihydro-6H-benzo[c]pyrido[1,2,3-lm]carbazole (2w), have been corroborated by single-crystal X-ray structural analyses. PMID- 21627123 TI - Comment on "Electronic structures, vibrational and thermochemical properties of neutral and charged niobium clusters Nb(n), n = 7-12". PMID- 21627124 TI - Elongation of phenoxide C-O bonds due to formation of multifold hydrogen bonds: statistical, experimental, and theoretical studies. AB - Statistical studies using the Cambridge Structural Database have revealed that there are several elongated phenoxide C-O bonds. They are characterized by the formation of 3-fold (or occasionally 2-fold) hydrogen bonds to the phenoxide oxygen atoms, and their mean bond length extends up to 1.320 A, which is quite different from the theoretically predicted carbon-oxygen bond length of C(6)H(5)O(-) (1.26 A). Elongated phenoxide C-O bonds associated with the formation of 3-fold hydrogen bonds were also observed in the X-ray structures of proton-transfer complexes (2X-O(-))(TEAH(+))s derived from 5'-X-substituted 5,5'' dimethyl-1,1':3',1''-terphenyl-2,2',2''-triols (2X-OHs, where X = NO(2), CN, COOCH(3), Cl, F, H, and CH(3)) and triethylamine (TEA). By comparing the X-ray structures, C-O bond elongation was found to be only slightly affected by an electron-withdrawing substituent at the para position (X). This along with strong bathochromic shifts of N-H(...O(-)) and O-H(...O(-)) stretching vibrations in the IR spectra indicates that the elongated C-O bonds in (2X-O(-))(TEAH(+))s essentially have single-bond character. This is further confirmed by molecular orbital calculations on a model complex, showing that the negatively charged phenoxide oxygen atom is no longer conjugated to the central benzene ring, and the NICS values of the three benzene rings are virtually identical. However, C-O bond elongation in (2X-O(-))(TEAH(+))s was considerably influenced by a change in the hydrogen-bond geometry. This also suggests that hydrogen bonds significantly affect phenoxide C-O bond elongation. PMID- 21627125 TI - Influence of the nature of the absorption band on the potential performance of high molar extinction coefficient ruthenium(II) polypyridinic complexes as dyes for sensitized solar cells. AB - When tested in solar cells, ruthenium polypyridinic dyes with extended pi systems show an enhanced light-harvesting capacity that is not necessarily reflected by a high (collected electrons)/(absorbed photons) ratio. Provided that metal-to ligand charge transfer bands, MLCT, are more effective, due to their directionality, than intraligand (IL) pi-pi* bands for the electron injection process in the solar cell, it seems important to explore and clarify the nature of the absorption bands present in these types of dyes. This article aims to elucidate if all the absorbed photons of these dyes are potentially useful in the generation of electric current. In other words, their potentiality as dyes must also be analyzed from the point of view of their contribution to the generation of excited states potentially useful for direct injection. Focusing on the assignment of the absorption bands and the nature of the emitting state, a systematic study for a series of ruthenium complexes with 4,4'-distyryl-2,2' dipyridine (LH) and 4,4'-bis[p-(dimethylamino)-alpha-styryl]-2,2'-bipyridine (LNMe(2)) "chromophoric" ligands was undertaken. The observed experimental results were complemented with TDDFT calculations to elucidate the nature of the absorption bands, and a theoretical model was proposed to predict the available energy that could be injected from a singlet or a triplet excited state. For the series studied, the results indicate that the percentage of MLCT character to the anchored ligand for the lower energy absorption band follows the order [Ru(deebpy)(2)(LNMe(2))](PF(6))(2) > [Ru(deebpy)(2)(LH)](PF(6))(2) > [Ru(deebpy)(LH)(2)](PF(6))(2), where deebpy is 4,4'-bis(ethoxycarbonyl)-2,2' bipyridine, predicting that, at least from this point of view, their efficiency as dyes should follow the same trend. PMID- 21627126 TI - Thermal and nonthermal physiochemical processes in nanoscale films of amorphous solid water. AB - Amorphous solid water (ASW) is a disordered version of ice created by vapor deposition onto a cold substrate (typically less than 130 K). It has a higher free energy than the crystalline phase of ice, and when heated above its glass transition temperature, it transforms into a metastable supercooled liquid. This unusual form of water exists on earth only in laboratories, after preparation with highly specialized equipment. It is thus fair to ask why there is any interest in studying such an esoteric material. Much of the scientific interest results from the ability to use ASW as a model system for exploring the physical and reactive properties of liquid water and aqueous solutions. ASW is also thought to be the predominant form of water in the extremely cold temperatures of many interstellar and planetary environments. In addition, ASW is a convenient model system for studying the stability of amorphous and glassy materials as well as the properties of highly porous materials. A fundamental understanding of such properties is invaluable in a diverse range of applications, including cryobiology, food science, pharmaceuticals, astrophysics, and nuclear waste storage, among others. Over the past 15 years, we have used molecular beams and surface science techniques to probe the thermal and nonthermal properties of nanoscale films of ASW. In this Account, we present a survey of our research on the properties of ASW using this approach. We use molecular beams to precisely control the deposition conditions (flux, incident energy, and incident angle) and create compositionally tailored, nanoscale films of ASW at low temperatures. To study the transport properties (viscosity and diffusivity), we heat the amorphous films above their glass transition temperature, T(g), at which they transform into deeply supercooled liquids prior to crystallization. The advantage of this approach is that at temperatures near T(g), the viscosity is approximately 15 orders of magnitude larger than that of a normal liquid. As a result, the crystallization kinetics are dramatically slowed, increasing the time available for experiments. For example, near T(g), a water molecule moves less than the distance of a single molecule on a typical laboratory time scale (~1000 s). For this reason, nanoscale films help to probe the behavior and reactions of supercooled liquids at these low temperatures. ASW films can also be used for investigating the nonthermal reactions relevant to radiolysis. PMID- 21627127 TI - Molecular simulation of aqueous electrolyte solubility. 2. Osmotic ensemble Monte Carlo methodology for free energy and solubility calculations and application to NaCl. AB - We present a new and computationally efficient methodology using osmotic ensemble Monte Carlo (OEMC) simulation to calculate chemical potential-concentration curves and the solubility of aqueous electrolytes. The method avoids calculations for the solid phase, incorporating readily available data from thermochemical tables that are based on well-defined reference states. It performs simulations of the aqueous solution at a fixed number of water molecules, pressure, temperature, and specified overall electrolyte chemical potential. Insertion/deletion of ions to/from the system is implemented using fractional ions, which are coupled to the system via a coupling parameter lambda that varies between 0 (no interaction between the fractional ions and the other particles in the system) and 1 (full interaction between the fractional ions and the other particles of the system). Transitions between lambda-states are accepted with a probability following from the osmotic ensemble partition function. Biasing weights associated with the lambda-states are used in order to efficiently realize transitions between them; these are determined by means of the Wang Landau method. We also propose a novel scaling procedure for lambda, which can be used for both nonpolarizable and polarizable models of aqueous electrolyte systems. The approach is readily extended to involve other solvents, multiple electrolytes, and species complexation reactions. The method is illustrated for NaCl, using SPC/E water and several force field models for NaCl from the literature, and the results are compared with experiment at ambient conditions. Good agreement is obtained for the chemical potential-concentration curve and the solubility prediction is reasonable. Future improvements to the predictions will require improved force field models. PMID- 21627128 TI - Near-IR laser-triggered target cell collection using a carbon nanotube-based cell cultured substrate. AB - Unique near-IR optical properties of single-walled carbon nanotube (SWNTs) are of interest in many biological applications. Here we describe the selective cell detachment and collection from an SWNT-coated cell-culture dish triggered by near IR pulse laser irradiation. First, HeLa cells were cultured on an SWNT-coated dish prepared by a spraying of an aqueous SWNT dispersion on a glass dish. The SWNT-coated dish was found to show a good cell adhesion behavior as well as a cellular proliferation rate similar to a conventional glass dish. We discovered, by near-IR pulse laser irradiation (at the laser power over 25 mW) to the cell under optical microscopic observation, a quick single-cell detachment from the SWNT-coated surface. Shockwave generation from the irradiated SWNTs is expected to play an important role for the cell detachment. Moreover, we have succeeded in catapulting the target single cell from the cultured medium when the depth of the medium was below 150 MUm and the laser power was stronger than 40 mW. The captured cell maintained its original shape. The retention of the genetic information of the cell was confirmed by the polymerase chain reaction (PCR) technique. A target single-cell collection from a culture medium under optical microscopic observation is significant in wide fields of single-cell studies in biological areas. PMID- 21627129 TI - Adlayer morphologies and free energy landscapes of clusters of bis-fullerenes on model gold surfaces. AB - There have been a few experimental reports of self-assembled adlayers of bis fullerene molecules on solid substrates. Most of these studies suggest the adsorbate molecules are lying down on the surface, with the fullerene moieties almost close packed. However, very little theoretical work has been carried out on such systems, and little is known about the roles played by different parts of the potential energy in driving the self-assembly. We carry out a Temperature Replica Exchange Monte Carlo study here of two representative bis-fullerene molecules on a metal substrate. We use a coarse-grained model potential energy function, in which certain parameters can be varied within the range of their experimental uncertainty. The molecules investigated consist of two fullerene moieties bonded by a rigid bridging group. In particular, the effect of the strength of the fullerene interaction E(FG) with the substrate (nominally Au(111)) has been investigated in detail. To ensure efficient sampling of the rugged potential energy surfaces encountered in the simulations, we utilize replica exchange techniques. These enable us to construct free energy landscapes for the system. We find that for relatively low values of E(FG) the molecules form standing-up adlayers. By contrast, for higher values of E(FG), lying-down adlayers dominate. For one molecule, two different crystalline adlayer morphologies have been identified. The detailed structure of the lying-down layer is a function of the temperature and of the group used to bridge the fullerene moieties. PMID- 21627130 TI - Features of Ar solvation shells in neutral and ionic clustering: the competitive role of two-body and many-body interactions. AB - The semiempirical methodology, introduced to describe noncovalent intermolecular interactions in atom/ion-molecule systems, is here extended to investigate a prototype cluster, formed by benzene (Bz) and closed-shell ions (Na(+) and/or Cl( )), surrounded by neutral species (Ar), forming solvation shells. The involved multidimensional potential energy surface (PES) is assumed to depend on a critical balancing of some effective interaction components. In particular, for the Ar solvated Bz-Na(+)-Cl(-) system, the nonelectrostatic component of the total interaction has been formulated as a combination of two-, three-, and four body contributions, each one represented by a proper function, with the four-body and part of the three-body terms arising from nonadditive induction effects. The proposed formulation, in which the induction is included both implicitly and explicitly, ensures the accurate description of all dissociation channels, leading to simpler clusters and/or pure solvent. Some properties of the solvent, represented by an ensemble of 500 Ar atoms, have been analyzed by performing molecular dynamics simulations at several temperatures. The obtained results have been found to be consistent with experimental observations. In order to investigate propensities, similarities, and differences in the competing clusters, the Ar solvation shells of Bz, Bz-Na(+), Bz-Cl(-) and Bz-Na(+)-Cl(-) have been characterized. The inspection of the solvation shell of Bz allows one to distinguish between groups of Ar atoms occupying positions on and out of the plane defined by the aromatic ring. Regarding the solvation shells of Bz-Na(+) and Bz-Cl(-), it has been observed that they are strongly affected by the most stable structures of the unsolvated systems. However, Bz-Na(+) shows more compact solvation shells than Bz-Cl(-). Finally, important asymmetries, basically promoted by the additional many-body induction effects on the solvent atoms, have been observed in the solvation shells of Bz-Na(+)-Cl(-). PMID- 21627131 TI - Colorimetric bacteria sensing using a supramolecular enzyme-nanoparticle biosensor. AB - Rapid and sensitive detection of pathogens is a key requirement for both environmental and clinical settings. We report here a colorimetric enzyme nanoparticle conjugate system for detection of microbial contamination. In this approach, cationic gold nanoparticles (NPs) featuring quaternary amine headgroups are electrostatically bound to an enzyme [beta-galactosidase (beta-Gal)], inhibiting enzyme activity. Analyte bacteria bind to the NP, which releases the beta-Gal and restores its activity, providing an enzyme-amplified colorimetric readout of the binding event. Using this strategy, we have been able to quantify bacteria at concentrations of 1 * 10(2) bacteria/mL in solution and 1 * 10(4) bacteria/mL in a field-friendly test strip format. PMID- 21627132 TI - D4(H)/D4(V) silicone: a replica material with several advantages for nanoimprint lithography and capillary force lithography. AB - Reported are demonstrations that D(4)(H)/D(4)(V) silicone (the product of the platinum-catalyzed hydrosilylation reaction between tetramethylcyclotetrasiloxane and tetramethyltetravinylcyclotetrasiloxane) is useful and practical as a replica material for both nanoimprint lithography (NIL) and capillary force lithography (CFL). The multiple advantageous properties of this extremely cross-linked material include UV transparency (for photo NIL and photo CFL), thermal stability (for high printing temperatures), high modulus (for high printing pressures), low surface energy (for easy demolding), and low viscosity precursors (for replicating small scale features). The replication performance of this material was tested using Blu-ray discs with sub-25 nm features and anodized aluminum foil with sub-10 nm features. Structures of ~5 nm length scale on the surface of the anodized Al were replicated using D(4)(H)/D(4)(V) silicone as a mold material for CFL with a photocurable epoxy resin and for NIL with poly(methyl methacrylate) (PMMA). Features (holes in the anodized aluminum) with aspect ratios of greater than 9 were replicated. PMID- 21627133 TI - Laser-induced nanoscale superhydrophobic structures on metal surfaces. AB - The combination of a dual-scale (nano and micro) roughness with an inherent low surface energy coating material is an essential factor for the development of superhydrophobic surfaces. Ultrashort pulse laser (USPL) machining/structuring is a promising technique for obtaining the dual-scale roughness. Sheets of stainless steel (AISI 304 L SS) and Ti-6Al-4V alloys were laser-machined with ultraviolet laser pulses of 6.7 ps, with different numbers of pulses per irradiated area. The surface energy of the laser-machined samples was reduced via application of a layer of perfluorinated octyltrichlorosilane (FOTS). The influence of the number of pulses per irradiated area on the geometry of the nanostructure and the wetting properties of the laser-machined structures has been studied. The results show that with an increasing number of pulses per irradiated area, the nanoscale structures tend to become predominantly microscale. The top surface of the microscale structures is seen covered with nanoscale protrusions that are most pronounced in Ti-6Al-4V. The laser-machined Ti-6Al-4V surface attained superhydrophobicity, and the improvement in the contact angle was >27% when compared to that of a nontextured surface. PMID- 21627134 TI - Chemoenzymatic asymmetric synthesis of optically active pentane-1,5-diamine fragments by means of lipase-catalyzed desymmetrization transformations. AB - A novel family of prochiral pentane-1,5-diamines has been efficiently synthesized, possessing stabilities significantly higher than those of corresponding propane-1,3-diamine analogues. Diamines have been later desymmetrized using Pseudomonas cepacia lipase as an efficient biocatalyst for the mono- but also stereoselective protection of one of their amino groups. Reaction parameters such as type and loading of enzyme, temperature, solvent, and acyl donor have been exhaustively analyzed, searching for optimal conditions for the production of interesting optically active nitrogenated compounds. Thus, acylation and alkoxycarbonylation processes have been compared in terms of conversion and enantiomeric excess values. The best results were found in the reaction of prochiral diamines with ethyl methoxyacetate as acyl donor and 1,4 dioxane as solvent, yielding (S)-monoamides in 33-59% isolated yield and 54-99% ee, depending on the aromatic pattern substitution. PMID- 21627135 TI - UV photolysis of alpha-cyclohexanedione in the gas phase. AB - Ultraviolet absorption spectrum of alpha-cyclohexanedione (alpha-CHD) vapor in the wavelength range of 220-320 nm has been recorded in a 1 m long path gas cell at room temperature. With the aid of theoretical calculation, the band has been assigned to the S(2) <- S(0) transition of largely pipi* type. The absorption cross section at the band maximum (~258 nm) is nearly 3 orders of magnitude larger compared to that for the S(2) <- S(0) transition of a linear alpha-diketo prototype, 2,3-pentanedione. The photolysis was performed by exciting the sample vapor near this band maximum, using the 253.7 nm line of a mercury vapor lamp, and the products were analyzed by mass spectrometry as well as by infrared spectroscopy. The identified products are cyclopentanone, carbon monoxide, ketene, ethylene, and 4-pentenal. Geometry optimization at the CIS/6-311++G** level predicts that the carbonyl group is pyramidally distorted in the excited S(1) and S(2) states, but the alpha-CHD ring does not show dissociative character. Potential energy curves with respect to a ring rupture coordinate (C-C bond between two carbonyl groups) for S(0), S(1), S(2), T(1), T(2), and T(3) states have been generated by partially optimizing the ground state geometry at DFT/B3LYP/6-311++G** level and calculating the vertical transition energies to the excited states by TDDFT method. Our analysis reveals that the reactions can take place at higher vibrational levels of S(0) as well as T(1) states. PMID- 21627136 TI - Nanocrystals of CeVO4 doped by metallic heteroions. AB - CeVO(4) nanocrystals doped by heteroions were prepared via a hydrothermal method without the presence of surfactants or templates. Transmission electron microscopy (TEM), high-resolution transmission electron microscopy (HRTEM), X-ray diffraction (XRD), solid state (51)V NMR, and inductively coupled plasma (ICP) were used to characterize the morphology, structure, and compositions of the materials. X-ray photoelectron spectroscopy (XPS) results confirmed that there is a valence change from Ce(3+) to Ce(4+) for a fraction of cerium atoms whereas the vanadium atoms remain in the pentavalent state V(5+) upon the substitution of Ca(2+) into CeVO(4). Raman spectroscopy was used to monitor the effects of the doping ion on the CeVO(4) lattice contraction and distortion. The appearance of the shifted and broadened Raman peaks for the doped CeVO(4) was interpreted by theoretical calculations performed with Vienna ab initio simulation package. The redox properties and photocatalytic activities of the obtained nanocrystals were also investigated and discussed in detail. PMID- 21627137 TI - Adhesion control for micro- and nanomanipulation. AB - The adhesion between a micro/nano-object and a microgripper end-effector is an important problem in micromanipulation. Canceling or reducing this force is a great challenge. This force is directly linked to the surface chemical structure of the object and the gripper. We propose to predict this force between a structuring surface and a micro-object with a multisphere van der Waals force model. The surface was structured by polystyrene latex particles (PS particles) with radii from 35 to 2000 nm. The model was compared with experimental pull-off force measurements performed by AFM with different natures of spheres materials glued on the tipless. A wide range of applications, in the field of telecommunications, bioengineering, and more generaly speaking MEMS can be envisaged for these substrates. PMID- 21627138 TI - Structural reorganization of a polymeric latex film during dry sintering at elevated temperatures. AB - Structural rearrangement in a latex powder during dry sintering at temperatures higher than the minimum film formation temperature was investigated by means of synchrotron small-angle X-ray scattering. Two major effects were identified: (1) Deformation of latex particles leads to a closure of voids between them and an extensive perfectioning of the face centered cubic colloidal crystalline ordering. Such an improvement of the colloidal crystalline structure involves preferential crystal growth along certain crystallographic directions as was evidenced by the measured unmatched relative diffraction intensity distribution of the crystallographic (111) and (220) planes. (2) Interdiffusion of polymeric chains between adjacent particles promotes a nanometer sized aggregation of nonpolymeric materials previously located in the interstices between particles. Size and size distribution of the aggregates at different dry sintering conditions were evaluated by using a model considering spheres dispersed in the system. PMID- 21627139 TI - Low-coordination sites in oxygen-reduction electrocatalysis: their roles and methods for removal. AB - Low-coordination sites, including edges, kinks, and defects, play an important role in oxygen-reduction electrocatalysis. Their role was studied experimentally and theoretically for various Pt surfaces. However, the roughness effect on similar-sized nanoparticles that could elucidate the role of low-coordination sites has attracted much less attention, with no studies on Pd nanoparticles. Here, using Br- adsorption/desorption, we introduce an effective approach to reduce surface roughness, yielding Pd nanoparticles with smoother surfaces and an increased number of (111)-oriented facets. The resulting nanoparticles have a slightly contracted structure and narrow size distribution. Pt monolayer catalysts that contain such nanoparticles as the cores showed a 1.5-fold enhancement in specific and Pt mass activities for the oxygen reduction reaction compared with untreated ones. Furthermore, a dramatic increase in durability was observed with bromide-treated Pd(3)Co cores. These results demonstrate a simple approach to preparing nanoparticles with smooth surfaces and confirm the adverse effect of low-coordination sites on the kinetics of the oxygen-reduction reaction. PMID- 21627140 TI - Columnarly assembled liquid-crystalline peptidic macrocycles unidirectionally orientable over a large area by an electric field. AB - Being inspired by naturally occurring peptidic macrocycles, we developed liquid crystalline (LC) compounds 1 and 2 that are capable of self-assembling into hexagonal columnar mesophases over a wide temperature range that includes room temperature. Their bowl-shaped macrocyclic cores are conformationally robust because of the presence of internal H-bonds, while the columnar assembly is ensured by intermolecular H-bonding interactions involving the exocyclic amide units. When an electric field was applied to their LC films from a direction orthogonal to the film plane, the columns were oriented homeotropically over a large area. PMID- 21627141 TI - Construction of a DOPC/PSM/cholesterol phase diagram based on the fluorescence properties of trans-parinaric acid. AB - Cell membranes have a nonhomogenous lateral organization. Most information about such nonhomogenous mixing has been obtained from model membrane studies where defined lipid mixtures have been characterized. Various experimental approaches have been used to determine binary and ternary phase diagrams for systems under equilibrium conditions. Such phase diagrams are the most useful tools for understanding the lateral organization in cellular membranes. Here we have used the fluorescence properties of trans-parinaric acid (tPA) for phase diagram determination. The fluorescence intensity, anisotropy, and fluorescence lifetimes of tPA were measured in bilayers composed of one to three lipid components. All of these parameters could be used to determine the presence of liquid-ordered and gel phases in the samples. However, the clearest information about the phase state of the lipid bilayers was obtained from the fluorescence lifetimes of tPA. This is due to the fact that an intermediate-length lifetime was found in samples that contain a liquid-ordered phase and a long lifetime was found in samples that contained a gel phase, whereas tPA in the liquid-disordered phase has a markedly shorter fluorescence lifetime. On the basis of the measured fluorescence parameters, a phase diagram for the 1,2-dioleoyl-sn-glycero-3-phosphocholine/N palmitoyl sphingomyelin/cholesterol system at 23 degrees C was prepared with a 5 mol % resolution. We conclude that tPA is a good fluorophore for probing the phase behavior of complex lipid mixtures, especially because multilamellar vesicles can be used. The determined phase diagram shows a clear resemblance to the microscopically determined phase diagram for the same system. However, there are also significant differences that likely are due to tPA's sensitivity to the presence of submicroscopic liquid-ordered and gel phase domains. PMID- 21627142 TI - Interaction of alkanethiols with nanoporous cluster-assembled Au films. AB - This article presents a study of the interaction of octadecanethiol molecules (C(18)) with nanoporous cluster-assembled gold films under a liquid environment based on a combined spectroscopic ellipsometry and X-ray photoelectron spectroscopy investigation. By comparing the optical response, following the deposition of C(18), of cluster-assembled films with varying degrees of porosity with that of flat surfaces and by resolving the corresponding features of the molecule-Au bond, we have been able to define the conditions that either favor molecular in-depth diffusion into the pores or promote the formation of a molecular self-assembled monolayer (SAM) restricted to the film surface. In the presence of abundant open pores, C(18) molecules strongly diffuse within the film interior and bind to the pore walls, whereas in the presence of porous films with less abundant open pores we have observed that the molecules tend to remain confined to the surface region, adopting a SAM-like configuration. PMID- 21627143 TI - Formation of assemblies comprising Ru-polypyridine complexes and CdSe nanocrystals studied by ATR-FTIR spectroscopy and DFT modeling. AB - The interaction between CdSe nanocrystals (NCs) passivated with trioctylphosphine oxide (TOPO) ligands and a series of Ru-polypyridine complexes [Ru(bpy)(3)](PF(6))(2) (1), [Ru(bpy)(2)(mcb)](PF(6))(2) (2), [Ru(bpy)(mcb)(2)](BarF)(2) (3), and [Ru(tpby)(2)(dcb)](PF(6))(2) (4) (where bpy = 2,2'-bipyridine, mcb = 4-carboxy-4'-methyl-2,2'-bipyridine, tbpy = 4,4'-di-tert butyl-2,2'-bipyridine; dcb = 4,4'-dicarboxy-2,2'-bipyridine, and BarF = tetrakis[3,5-bis(trifluoromethyl)phenyl]borate)-was studied by attenuated total reflectance FTIR (ATR-FTIR) and modeled using density functional theory (DFT). ATR-FTIR studies reveal that when the solid film of NCs is exposed to an acetonitrile solution of 2, 3, or 4, the complexes chemically bind to the NC surface through their carboxylic acid groups, replacing TOPO ligands. The corresponding spectral changes are observed on a time scale of minutes. In the case of 2, the FTIR spectral changes clearly show that the complex adsorption is associated with a loss of proton from the carboxylic acid group. In the case of 3 and 4, deprotonation of the anchoring group is also detected, while the second, "spectrator" carboxylic acid group remains protonated. The observed energy difference between the symmetric, nu(s), and asymmetric, nu(as), stretch of the deprotonated carboxylic acid group suggests that the complexes are bound to the NC surface via a bridging mode. The results of DFT modeling are consistent with the experiment, showing that for the deprotonated carboxylic acid group the coupling to two Cd atoms via a bridging mode is the energetically most favorable mode of attachment for all nonequivalent NC surface sites and that the attachment of the protonated carboxylic acid is thermodynamically significantly less favorable. PMID- 21627144 TI - Dynamics of droplet motion under electrowetting actuation. AB - The static shape of droplets under electrowetting actuation is well understood. The steady-state shape of the droplet is obtained on the basis of the balance of surface tension and electrowetting forces, and the change in the apparent contact angle is well characterized by the Young-Lippmann equation. However, the transient droplet shape behavior when a voltage is suddenly applied across a droplet has received less attention. Additional dynamic frictional forces are at play during this transient process. We present a model to predict this transient behavior of the droplet shape under electrowetting actuation. The droplet shape is modeled using the volume of fluid method. The electrowetting and dynamic frictional forces are included as an effective dynamic contact angle through a force balance at the contact line. The model is used to predict the transient behavior of water droplets on smooth hydrophobic surfaces under electrowetting actuation. The predictions of the transient behavior of droplet shape and contact radius are in excellent agreement with our experimental measurements. The internal fluid motion is explained, and the droplet motion is shown to initiate from the contact line. An approximate mathematical model is also developed to understand the physics of the droplet motion and to describe the overall droplet motion and the contact line velocities. PMID- 21627145 TI - Beneficial effects of dietary fish-oil-derived monounsaturated fatty acids on metabolic syndrome risk factors and insulin resistance in mice. AB - The aim of this study was to elucidate the effect of fish-oil-derived monounsaturated fatty acids (MUFAs) containing large amounts of C20:1 and C22:1 isomers on metabolic disorders in mice. Male C57BL/6J mice were fed a 32% lard diet (control) or a 27% lard plus 5% saury-oil-derived MUFA diet for 6 weeks. Dietary MUFA improved insulin resistance and alleviated metabolic syndrome risk factors by reducing blood glucose and lipids. These favorable changes may be attributed to an improved adipocytokine profile. MUFA ingestion resulted in favorable changes in mRNA expression of genes involved in glucose/lipid metabolism (SCD-1, CPT1a, UCPs, and CS) as well as inflammation (MAC1, MMP3, and SAA3) and alterations in fatty acid composition. Our data suggest that marine MUFA improved glucose/lipid homeostasis and hindered the development of metabolic syndrome in obese mice. PMID- 21627147 TI - Determination of physicochemical parameters of ionic liquids and their mixtures with solvents using laser-induced gratings. AB - The laser-induced gratings (LIGs) technique has been applied for the simultaneous determination of sound speed and thermal diffusivity in the room temperature ionic liquid (RTIL) 1-ethyl-3-methylimidazolium ethylsulfate, [EMIm][EtSO(4)], its mixture with 85.68 mol % acetone, C(3)H(6)O, and pure acetone. The measurements have been performed in a quartz glass cuvette at ambient pressure and temperature. Radiation of a pulse-repetitive Q-switched Nd:YAG pump laser (1064 nm) effected quasi-resonant excitation of overtone-combinational vibrational states of the RTIL molecules followed by the appearance of laser induced gratings. The temporal evolution of the transient gratings (oscillation and damping) was recorded using Bragg-diffraction of a continuous-wave probe laser radiation. From the LIG signals' temporal profiles, values of the sound speed and thermal diffusivity were determined and, in addition, the isentropic compressibility and thermal conductivity were derived. The results are in a reasonable agreement with those reported in the literature. Furthermore, since the data for the determination of the physicochemical properties can be obtained with a single laser pulse, the LIG technique has potential for applications where data acquisition at high repetition rates is desirable for example to monitor processes. PMID- 21627148 TI - Reaction kinetics of CO2 carbonation with Mg-rich minerals. AB - Due to their low price, wide availability, and stability of the resulting carbonates, Mg-rich minerals are promising materials for carbonating CO(2). Direct carbonation of CO(2) with Mg-rich minerals reported in this research for the first time could be considerably superior to conventional liquid extraction processes from an energy consumption perspective due to its avoidance of the use of a large amount of water with high specific heat capacity and latent heat of vaporization. Kinetic models of the reactions of the direct CO(2) carbonation with Mg-rich minerals and within simulated flue gas environments are important to the scale-up of reactor designs. Unfortunately, such models have not been made available thus far. This research was initiated to fill that gap. Magnesium silicate (Mg(2)SiO(4)), a representative compound in Mg-rich minerals, was used to study CO(2) carbonation reaction kinetics under given simulated flue gas conditions. It was found that the chosen sorbent deactivation model fits well the experimental data collected under given conditions. A reaction order of 1 with respect to CO(2) is obtained from experimental data. The Arrhenius form of CO(2) carbonation with Mg(2)SiO(4) is established based on changes in the rate constants of the chosen deactivation model as a function of temperature. PMID- 21627146 TI - Complexes formed in solution between vanadium(IV)/(V) and the cyclic dihydroxamic acid putrebactin or linear suberodihydroxamic acid. AB - An aerobic solution prepared from V(IV) and the cyclic dihydroxamic acid putrebactin (pbH(2)) in 1:1 H(2)O/CH(3)OH at pH = 2 turned from blue to orange and gave a signal in the positive ion electrospray ionization mass spectrometry (ESI-MS) at m/z(obs) 437.0 attributed to the monooxoV(V) species [V(V)O(pb)](+) ([C(16)H(26)N(4)O(7)V](+), m/z(calc) 437.3). A solution prepared as above gave a signal in the (51)V NMR spectrum at delta(V )= -443.3 ppm (VOCl(3), delta(V) = 0 ppm) and was electron paramagnetic resonance silent, consistent with the presence of [V(V)O(pb)](+). The formation of [V(V)O(pb)](+) was invariant of [V(IV)]:[pbH(2)] and of pH values over pH = 2-7. In contrast, an aerobic solution prepared from V(IV) and the linear dihydroxamic acid suberodihydroxamic acid (sbhaH(4)) in 1:1 H(2)O/CH(3)OH at pH values of 2, 5, or 7 gave multiple signals in the positive and negative ion ESI-MS, which were assigned to monomeric or dimeric V(V)- or V(IV)-sbhaH(4) complexes or mixed-valence V(V)/(IV)-sbhaH(4) complexes. The complexity of the V-sbhaH(4) system has been attributed to dimerization (2[V(V)O(sbhaH(2))](+) <-> [(V(V)O)(2)(sbhaH(2))(2)](2+)), deprotonation ([V(V)O(sbhaH(2))](+) - H(+) <-> [V(V)O(sbhaH)](0)), and oxidation ([V(IV)O(sbhaH(2))](0) -e(-) <-> [V(V)O(sbhaH(2))](+)) phenomena and could be described as the sum of two pH-dependent vectors, the first comprising the deprotonation of hydroxamate (low pH) to hydroximate (high pH) and the second comprising the oxidation of V(IV) (low pH) to V(V) (high pH). Macrocyclic pbH(2) was preorganized to form [V(V)O(pb)](+), which would provide an entropy-based increase in its thermodynamic stability compared to V(V)-sbhaH(4) complexes. The half-wave potentials from solutions of [V(IV)]:[pbH(2)] (1:1) or [V(IV)]:[sbhaH(4)] (1:2) at pH = 2 were E(1/2) -335 or -352 mV, respectively, which differed from the expected trend (E(1/2) [VO(pb)](+/0) < V(V/IV)-sbhaH(4)). The complex solution speciation of the V(V)/(IV)-sbhaH(4) system prevented the determination of half-wave potentials for single species. The characterization of [V(V)O(pb)](+) expands the small family of documented V-siderophore complexes relevant to understanding V transport and assimilation in the biosphere. PMID- 21627149 TI - Electrophoresis of a charged droplet in a dielectric liquid for droplet actuation. AB - Electrophoretic motion of a charged droplet in a dielectric fluid under an electric field has been investigated experimentally for use as a microdroplet actuation method. The effects of the droplet size, electric field strength, and electrolyte concentration and ion species on the charging of an aqueous droplet have been examined. The amount of electrical charging has been measured by two different methods: indirect measurement using the image analysis of droplet motion and direct measurement using the electrometer. Quantitative comparison of the droplet charge measured experimentally and the theoretical value of a perfectly conductive sphere shows that an aqueous droplet is less charged than the corresponding perfectly conductive sphere. The limiting effect on electrical charging is more significant for an electrolyte droplet, and the effect is positively correlated to the electrolyte concentration rather than the ion species. This implies that the low electrical conductivity of water is not a major cause of the limiting effect. The scaling law of the charging amount for a deionized water droplet nearly follows that of the perfect conductor, whereas for an electrolyte droplet, the scaling law exponent is slightly higher. Some advantages and potentials of the current droplet actuation method are also discussed in comparison with the conventional ones. PMID- 21627150 TI - New oxides showing an intense orange color based on Fe3+ in trigonal-bipyramidal coordination. AB - Hexagonal YIn(1-x)Fe(x)O(3) phases have been prepared and characterized. The coordination for the In/Fe site in this structure is trigonal-bipyramidal. The colors of the phases change from yellow to orange to dark red with increasing Fe content. Magnetic measurements confirm high-spin Fe(3+) for all phases. PMID- 21627151 TI - Combining the pair distribution function and computational methods to understand lithium insertion in Brookite (TiO2). AB - X-ray pair distribution function (PDF) methods and first-principles calculations have been combined to probe the structure of electrochemically lithiated TiO(2) Brookite. Traditional powder diffraction studies suggest that Brookite amorphizes upon lithium insertion, with the Bragg reflections disappearing. However, PDF analysis indicates that the TiO(2) framework connectivity is maintained throughout lithium intercalation, with expansions along the a and b axes. The Li(+) ions within the framework are poorly observed in the X-ray PDF, which is dominated by contributions from the more strongly scattering Ti and O atoms. First-principles calculations were used to identify energetically favorable Li(+) sites within the Brookite lattice and to develop a complete structural model of the lithiated material. This model replicates the local structure and decreased intermediate range order observed in the PDF data. The analysis suggests that local structural distortions of the TiO(2) lattice accommodate lithium in five coordinate sites. This structural model is consistent with the observed electrochemical behavior. PMID- 21627152 TI - Low-valent ene-amido iron complexes for the asymmetric transfer hydrogenation of acetophenone without base. AB - Examination of the role of base in the activation of our previously reported iron(II) complexes having the general formula [Fe(CO)(Br)(PNNP)][BPh(4)] revealed a five-coordinate iron(II) complex in which the tetradentate PNNP ligand had been doubly deprotonated. The new iron(II) complexes were used in the transfer hydrogenation of acetophenone in isopropanol in the absence of added base, and certain analogues showed catalytic activity. PMID- 21627153 TI - Mobility of solid tert-butyl alcohol studied by deuterium NMR. AB - The molecular mobility of solid deuterated tert-butyl alcohol (TBA) has been studied over a broad temperature range (103-283 K) by means of solid-state 2H NMR spectroscopy, including both line shape and anisotropy of spin-lattice relaxation analyses. It has been found that, while the hydroxyl group of the TBA molecule is immobile on the 2H NMR time scale (tauC > 10(-5) s), its butyl group is highly mobile. The mobility is represented by the rotation of the methyl [CD3] groups about their 3-fold axes (C3 rotational axis) and the rotation of the entire butyl [(CD3)3-C] fragment about its 3-fold axis (C3' rotational axis). Numerical simulations of spectra line shapes reveal that the methyl groups and the butyl fragment exhibit three-site jump rotations about their symmetry axes C3 and C3' in the temperature range of 103-133 K, with the activation energies and preexponential factors E1 = 21 +/- 2 kJ/mol, k(01) = (2.6 +/- 0.5) * 10(12) s(-1) and E2 = 16 +/- 2 kJ/mol, k(02) = (1 +/- 0.2) * 10(12) s(-1), respectively. Analysis of the anisotropy of spin-lattice relaxation has demonstrated that the reorientation mechanism of the butyl fragment changes to a free diffusion rotational mechanism above 173 K, while the rotational mechanism of the methyl groups remains the same. The values of the activation barriers for both rotations at T > 173 K have the values, which are similar to those at 103-133 K. This indicates that the interaction potential defining these motions remains unchanged. The obtained data demonstrate that the detailed analysis of both line shape and anisotropy of spin-lattice relaxation represents a powerful tool to follow the evolution of the molecular reorientation mechanisms in organic solids. PMID- 21627154 TI - Asymmetric Strecker synthesis of alpha-arylglycines. AB - A practically simple three-component Strecker reaction for the asymmetric synthesis of enantiopure alpha-arylglycines has been developed. Addition of a range of aryl-aldehydes to a solution of sodium cyanide and (S)-1-(4 methoxyphenyl)ethylamine affords highly crystalline (S,S)-alpha-aminonitriles that are easily obtained in diastereomerically pure form. Heating the resultant (S,S)-alpha-aminonitriles in 6 M aqueous HCl at reflux resulted in cleavage of their chiral auxiliary fragments and concomitant hydrolysis of their nitrile groups to afford enantiopure (S)-alpha-arylglycines. The enantiopurities of these (S)-alpha-arylglycines were determined via derivatization of their corresponding methyl esters with 2-formylphenylboronic acid and (S)-BINOL, followed by (1)H NMR spectroscopic analysis of the resultant mixtures of diastereomeric iminoboronate esters. PMID- 21627155 TI - Contribution of extracellular polymeric substances from Shewanella sp. HRCR-1 biofilms to U(VI) immobilization. AB - The goal of this study was to quantify the contribution of extracellular polymeric substances (EPS) to U(VI) immobilization by Shewanella sp. HRCR-1. Through comparison of U(VI) immobilization using cells with bound EPS (bEPS) and cells with minimal EPS, we show that (i) bEPS from Shewanella sp. HRCR-1 biofilms contribute significantly to U(VI) immobilization, especially at low initial U(VI) concentrations, through both sorption and reduction; (ii) bEPS can be considered a functional extension of the cells for U(VI) immobilization and they likely play more important roles at lower initial U(VI) concentrations; and (iii) the U(VI) reduction efficiency is dependent upon the initial U(VI) concentration and decreases at lower concentrations. To quantify the relative contributions of sorption and reduction to U(VI) immobilization by EPS fractions, we isolated loosely associated EPS (laEPS) and bEPS from Shewanella sp. HRCR-1 biofilms grown in a hollow fiber membrane biofilm reactor and tested their reactivity with U(VI). We found that, when reduced, the isolated cell-free EPS fractions could reduce U(VI). Polysaccharides in the EPS likely contributed to U(VI) sorption and dominated the reactivity of laEPS, while redox active components (e.g., outer membrane c-type cytochromes), especially in bEPS, possibly facilitated U(VI) reduction. PMID- 21627156 TI - Formal total synthesis of (-)-oseltamivir phosphate. AB - An asymmetric synthesis of chiral intermediate 3 for (-)-oseltamivir phosphate has been accomplished from chiral building block 1, which was prepared by catalytic asymmetric synthesis. PMID- 21627157 TI - The short-lived signaling state of the photoactive yellow protein photoreceptor revealed by combined structural probes. AB - The signaling state of the photoactive yellow protein (PYP) photoreceptor is transiently developed via isomerization of its blue-light-absorbing chromophore. The associated structural rearrangements have large amplitude but, due to its transient nature and chemical exchange reactions that complicate NMR detection, its accurate three-dimensional structure in solution has been elusive. Here we report on direct structural observation of the transient signaling state by combining double electron electron resonance spectroscopy (DEER), NMR, and time resolved pump-probe X-ray solution scattering (TR-SAXS/WAXS). Measurement of distance distributions for doubly spin-labeled photoreceptor constructs using DEER spectroscopy suggests that the signaling state is well ordered and shows that interspin-label distances change reversibly up to 19 A upon illumination. The SAXS/WAXS difference signal for the signaling state relative to the ground state indicates the transient formation of an ordered and rearranged conformation, which has an increased radius of gyration, an increased maximum dimension, and a reduced excluded volume. Dynamical annealing calculations using the DEER derived long-range distance restraints in combination with short-range distance information from (1)H-(15)N HSQC perturbation spectroscopy give strong indication for a rearrangement that places part of the N-terminal domain in contact with the exposed chromophore binding cleft while the terminal residues extend away from the core. Time-resolved global structural information from pump probe TR-SAXS/WAXS data supports this conformation and allows subsequent structural refinement that includes the combined energy terms from DEER, NMR, and SAXS/WAXS together. The resulting ensemble simultaneously satisfies all restraints, and the inclusion of TR-SAXS/WAXS effectively reduces the uncertainty arising from the possible spin-label orientations. The observations are essentially compatible with reduced folding of the I(2)' state (also referred to as the 'pB' state) that is widely reported, but indicates it to be relatively ordered and rearranged. Furthermore, there is direct evidence for the repositioning of the N-terminal region in the I(2)' state, which is structurally modeled by dynamical annealing and refinement calculations. PMID- 21627158 TI - Selective catalytic reduction at quasi-perfect Pt(100) domains: a universal low temperature pathway from nitrite to N2. AB - The highly selective conversion of nitrite to N(2) at a quasi-perfect Pt(100) electrode in alkaline media was investigated with a particular emphasis on its structure sensitivity and its mechanism. High-quality (100) terraces are required to optimize the catalytic activity and steer the selectivity to N(2): defects of any symmetry dramatically reduce the N(2) evolution at [(100) * (110)] and [(100) * (111)] surfaces. On the other hand, nitrite reduction proves to be an additional example of the unique intrinsic ability of (100) surfaces to catalyze reactions involving bond breaking and successive bond formation. In the present case, (100) is able to reduce nitrite to NH(2,ads), which in a certain potential window combines with NO(ads) to give N(2) in a Langmuir-Hinshelwood reaction. Our findings are similar to those for other processes generating N(2), from bacterial anoxic ammonia oxidation ("anammox") to the high-temperature NO + NH(3) reaction at Pt(100) crystals under ultra-high-vacuum conditions, thus suggesting that the combination of these two nitrogen-containing species is a universal (low temperature) pathway to N(2). The advantages of this pathway over other N(2) generating pathways are pointed out. PMID- 21627159 TI - Physicochemical characterization of particulate emissions from a compression ignition engine employing two injection technologies and three fuels. AB - Alternative fuels and injection technologies are a necessary component of particulate emission reduction strategies for compression ignition engines. Consequently, this study undertakes a physicochemical characterization of diesel particulate matter (DPM) for engines equipped with alternative injection technologies (direct injection and common rail) and alternative fuels (ultra low sulfur diesel, a 20% biodiesel blend, and a synthetic diesel). Particle physical properties were addressed by measuring particle number size distributions, and particle chemical properties were addressed by measuring polycyclic aromatic hydrocarbons (PAHs) and reactive oxygen species (ROS). Particle volatility was determined by passing the polydisperse size distribution through a thermodenuder set to 300 degrees C. The results from this study, conducted over a four point test cycle, showed that both fuel type and injection technology have an impact on particle emissions, but injection technology was the more important factor. Significant particle number emission (54%-84%) reductions were achieved at half load operation (1% increase-43% decrease at full load) with the common rail injection system; however, the particles had a significantly higher PAH fraction (by a factor of 2 to 4) and ROS concentrations (by a factor of 6 to 16) both expressed on a test-cycle averaged basis. The results of this study have significant implications for the health effects of DPM emissions from both direct injection and common rail engines utilizing various alternative fuels. PMID- 21627160 TI - Pollutant formation and emissions from cement kiln stack using a solid recovered fuel from municipal solid waste. AB - The thermal decomposition of a solid recovered fuel (SRF) has been studied by two techniques. First, laboratory-scale experiments were performed in a horizontal furnace in which different atmospheres were studied to analyze the dioxins and furans (PCDD/Fs) evolved from the decomposition of the material. Sulfur presence was revealed to be important in PCDD/Fs formation. In the second technique, the emissions of various pollutants (PAHs, PCDD/Fs, metals, acid gases, etc.) were determined in a cement kiln fed on different proportions of SRF material, and where a maximum feed rate of 15000 kg SRF/h was achieved. In the laboratory furnace the dioxin toxicity revealed a maximum when the amount of oxygen in the atmosphere increased until approximately stoichiometric conditions. In the cement kiln, all emitted pollutants were under the legal limits. No correlation between SRF input and metal emission was observed. PMID- 21627161 TI - Conformational flexibility of a human immunoglobulin light chain variable domain by relaxation dispersion nuclear magnetic resonance spectroscopy: implications for protein misfolding and amyloid assembly. AB - The conformational flexibility of a human immunoglobulin kappaIV light-chain variable domain, LEN, which can undergo conversion to amyloid under destabilizing conditions, was investigated at physiological and acidic pH on a residue-specific basis by multidimensional solution-state nuclear magnetic resonance (NMR) methods. Measurements of backbone chemical shifts and amide (15)N longitudinal and transverse spin relaxation rates and steady-state nuclear Overhauser enhancements indicate that, on the whole, LEN retains its native three dimensional fold and dimeric state at pH 2 and that the protein backbone exhibits limited fast motions on the picosecond to nanosecond time scale. On the other hand, (15)N Carr--Purcell--Meiboom--Gill (CPMG) relaxation dispersion NMR data show that LEN experiences considerable slower, millisecond time scale dynamics, confined primarily to three contiguous segments of about 5-20 residues and encompassing the N-terminal beta-strand and complementarity determining loop regions 2 and 3 in the vicinity of the dimer interface. Quantitative analysis of the CPMG relaxation dispersion data reveals that at physiological pH these slow backbone motions are associated with relatively low excited-state protein conformer populations, in the ~2-4% range. Upon acidification, the minor conformer populations increase significantly, to ~10-15%, with most residues involved in stabilizing interactions across the dimer interface displaying increased flexibility. These findings provide molecular-level insights about partial protein unfolding at low pH and point to the LEN dimer dissociation, initiated by increased conformational flexibility in several well-defined regions, as being one of the important early events leading to amyloid assembly. PMID- 21627162 TI - Antioxidants in foods: state of the science important to the food industry. AB - Antioxidant foods and ingredients are an important component of the food industry. In the past, antioxidants were used primarily to control oxidation and retard spoilage, but today many are used because of putative health benefits. However, the traditional message that oxidative stress, which involves the production of reactive oxygen species (ROS), is the basis for chronic diseases and aging is being reexamined. Accumulating evidence suggests that ROS exert essential metabolic functions and that removal of too many ROS can upset cell signaling pathways and actually increase the risk of chronic disease. It is imperative that the food industry be aware of progress in this field to present the science relative to foods in a forthright and clear manner. This may mean reexamining the health implications of adding large amounts of antioxidants to foods. PMID- 21627163 TI - Microscale technologies and modular approaches for tissue engineering: moving toward the fabrication of complex functional structures. AB - Micro- and nanoscale technologies have emerged as powerful tools in the fabrication of engineered tissues and organs. Here we focus on the application of these techniques to improve engineered tissue architecture and function using modular and directed self-assembly and highlight the emergence of this new class of materials for biomedical applications. PMID- 21627164 TI - Theoretical and experimental study of weakly bound CO2-(pH2)2 trimers. AB - The infrared spectrum of CO(2)-(pH(2))(2) trimers is predicted by performing exact basis-set calculations on a global potential energy surface defined as the sum of accurately known two-body pH(2)-CO(2) (J. Chem. Phys. 2010, 132, 214309) and pH(2)-pH(2) potentials (J. Chem. Phys. 2008, 129, 094304). These results are compared with new spectroscopic measurements for this species, for which 13 transitions are now assigned. A reduced-dimension treatment of the pH(2) rotation has been employed by applying the hindered-rotor averaging technique of Li, Roy, and Le Roy (J. Chem. Phys. 2010, 133, 104305). Three-body effects and the quality of the potential are discussed. A new technique for displaying the three dimensional pH(2) density in the body-fixed frame is used, and shows that in the ground state the two pH(2) molecules are localized much more closely together than is the case for the two He atoms in the analogous CO(2)-(He)(2) species. A clear tunneling splitting is evident for the torsional motion of the two pH(2) molecules on a ring about the CO(2) molecular axis, in contrast to the case of CO(2)-(He)(2) where a more regular progression of vibrational levels reflects the much lower torsional barrier. PMID- 21627165 TI - Model simulations of particle aggregation effect on colloid exchange between streams and streambeds. AB - Colloids found in natural streams have large reactive surface areas, which makes them significant absorbents and carriers for pollutants. Stream-subsurface exchange plays a critical role in regulating the transport of colloids and contaminants in natural streams. Previous process-based multiphase exchange models were developed without consideration of colloid-colloid interaction. However, many studies have indicated that aggregation is a significant process and needs to be considered in stream process analysis. Herein, a new colloid exchange model was developed by including particle aggregation in addition to colloid settling and filtration. Self-preserving size distribution concepts and classical aggregation theory were employed to model the aggregation process. Model simulations indicate that under conditions of low filtration and high degree of particle-particle interaction, aggregation could either decrease or increase the amount of colloids retained in streambeds, depending on the initial particle size. Thus, two possible cases may occur including enhanced colloid deposition and facilitated colloid transport. Also, when the aggregation rate is high and filtration increases, more particles are retained by bed sediments due to filtration, and fewer are aggregated, which reduces the extent of aggregation effect on colloid deposition. The work presented here will contribute to a better understanding and prediction of colloid transport phenomena in natural streams. PMID- 21627166 TI - Molecular dynamics simulation study of ion interactions with zwitterions. AB - Using molecular dynamics simulations, we investigated the associations between two zwitterions (carboxybetaine and sulfobetaine) and four types of cations (Li(+), Na(+), K(+), and Cs(+)) in aqueous solutions. We studied the number and lifetime of various zwitterion-cation associations and observed that both carboxybetaine and sulfobetaine have the same order of association number and lifetime: Li(+) > Na(+) > K(+) > Cs(+). Simulation results showed that the association variation as a function of cation types for these two zwitterions is significantly different. The effect of anion type on the order was also investigated by varying the type of anions from Cl(-) to Br(-) and F(-). In order to further investigate zwitterion-cation association, we simulated the systems either with one type of zwitterion and two types of cations or with one type of cation and both carboxybetaine and sulfobetaine presented. This allowed direct competition between the solutes, and the observed association number and lifetime validated the order. Simulation results further demonstrated that, although CB associates stronger with Li(+) and Na(+) than SB, the latter is associated preferentially by K(+) and Cs(+). PMID- 21627168 TI - Solvent effects on chemical exchange in a push-pull ethylene as studied by NMR and electronic structure calculations. AB - NMR measurements of chemical exchange in a push-pull ethylene, dissolved in a number of different solvents, are presented. These are complemented by high-level electronic structure calculations, using both gas-phase conditions and those which simulate solvents. The results show that it is essential to include entropy effects in order to understand the observed trends. For instance, the equilibrium state in this case represents the state with lowest Gibbs free energy, as it must, but not the lowest enthalpy. The particular molecule is methyl 3 dimethylamino-2-cyanocrotonate (MDACC). The geometry at the carbon-carbon double bond can be either E or Z with roughly equal populations at ambient temperature. We have measured the equilibrium constant and the rates for the exchange between these states in a number of solvents: methanol, chloroform, acetonitrile, toluene, dichloromethane, acetone, and tetrahydrofuran. Furthermore, the N,N dimethylamino group attached to the double bond also shows restricted rotation, and this has been measured in both the E and Z conformations. The equilibrium constant and the three rotational barriers provide excellent probes of the solvent effects. Electronic structure calculations with a number of basis sets up to the 6-311++G(2df,2p) level, using both Hartree-Fock and density functional (B3LYP) methods were used to predict the E and Z ground states, and the three transition states. The calculations were done for an isolated molecule and also for solvent models representing toluene, acetone, and ethanol. The E conformation is more stable in solution, is the structure in the crystal, and is also the prediction for the gas phase from the calculations. However, the dependence of the equilibrium constant on temperature shows that the Z conformation actually has lower enthalpy. The stability of the E conformation in solution must be due to entropic effects. Similarly, the solvent effect on the E-Z barrier is primarily due to entropy. The measured enthalpy of activation is similar in all the solvents, but the entropy of activation increases with the solvent polarity. The barrier to rotation of the N,N-dimethylamino group shows a combination of entropy and enthalpy effects. This combination of experiments and theory gives an extraordinarily detailed picture of solvent-solute interactions. PMID- 21627167 TI - Water-mediated binding of agents that target the DNA minor groove. AB - Small molecule complexes with DNA that incorporate linking water molecules are rare, and the DB921-DNA complex has provided a unique and well-defined system for analysis of water-mediated binding in the context of a DNA complex. DB921 has a benzimidazole-biphenyl system with terminal amidines that results in a linear conformation that does not possess the appropriate radius of curvature to match the minor groove shape and represents a new paradigm that does not fit the classical model of minor groove interactions. To better understand the role of the bound water molecule observed in the X-ray crystal structure of the DB921 complex, synthetic modifications have been made in the DB921 structure, and the interactions of the new compounds with DNA AT sites have been evaluated with an array of methods, including DNase I footprinting, biosensor-surface plasmon resonance, isothermal titration microcalorimetry, and circular dichroism. The interaction of a key compound, which has the amidine at the phenyl shifted from the para position in DB921 to the meta position, has also been examined by X-ray crystallography. The detailed structural, thermodynamic, and kinetic results provide valuable new information for incorporation of water molecules in the design of new lead scaffolds for targeting DNA in chemical biology and therapeutic applications. PMID- 21627169 TI - Asymmetric [C + NC + CC] coupling entry to the naphthyridinomycin natural product family: formal total synthesis of cyanocycline A and bioxalomycin beta2. AB - A full account of our [C + NC + CC] coupling approach to the naphthyridinomycin family of natural products is presented, culminating in formal total syntheses of cyanocycline A and bioxalomycin beta2. The key complexity-building reaction in the synthesis involves the Ag(I)-catalyzed endo-selective [C + NC + CC] coupling of aldehyde 7, (S)-glycyl sultam 8, and methyl acrylate (9) to provide the highly functionalized pyrrolidine 6, which was carried forward to an advanced intermediate (compound 33) in Fukuyama's synthesis of cyanocycline A. Since cyanocycline A has been converted to bioxalomycin beta2, this constitutes a formal synthesis of the latter natural product as well. The multicomponent reaction-based strategy reduces the number of steps previously needed to assemble these complex molecular targets by one-third. This work highlights the utility of the asymmetric [C + NC + CC] coupling reaction in the context of a complex pyrrolidine-containing target and provides an illustrative guide for its application to other synthesis problems. The synthesis also fueled collaborative biological and biochemical research that identified a unique small molecule inhibitor of cell migration (compound 30). PMID- 21627170 TI - Toward a four-toothed molecular bevel gear with C2-symmetrical rotors. AB - The design, synthesis, conformational analysis, and variable-temperature NMR studies of pentiptycene-based molecular gears Pp(2)X, where Pp is the unlabeled (in 1H) or methoxy groups-labeled (in 1OM) pentiptycene rotor and X is the phenylene stator containing ortho-bridged ethynylene axles, are reported. The approach of using shape-persistent rotors of four teeth but C(2) symmetry for constructing four-toothed molecular gears is unprecedented. In addition, the first example of enantioresolution of chiral pentiptycene scaffolds is demonstrated. Density functional theory (DFT) and AM1 calculations on these Pp(2)X systems suggest two possible correlated torsional motions, geared rocking and four-toothed geared rotations, which compete with the uncorrelated gear slippage. The DFT-derived torsional barriers in 1H for rocking, four-toothed rotation, and gear slippage are approximately 2.9, 5.5, and 4.7 kcal mol(-1), respectively. The low energy barriers for these torsional motions result from the low energy cost of bending the ethynylene axles. Comparison of the NMR spectra of 1OM in a mixture of stereoisomers (1OM-mix) and in an enantiopure form (1OM-op) confirms a fast gear slippage in these Pp(2)X systems. The effect of the methoxy labels on rotational potential energy surface and inter-rotor dynamics is also discussed. PMID- 21627171 TI - Intramolecular and intermolecular contributions to the barriers for rotation of methyl groups in crystalline solids: electronic structure calculations and solid state NMR relaxation measurements. AB - The rotation barriers for 10 different methyl groups in five methyl-substituted phenanthrenes and three methyl-substituted naphthalenes were determined by ab initio electronic structure calculations, both for the isolated molecules and for the central molecules in clusters containing 8-13 molecules. These clusters were constructed computationally using the carbon positions obtained from the crystal structures of the eight compounds and the hydrogen positions obtained from electronic structure calculations. The calculated methyl rotation barriers in the clusters (E(clust)) range from 0.6 to 3.4 kcal/mol. Solid-state (1)H NMR spin lattice relaxation rate measurements on the polycrystalline solids gave experimental activation energies (E(NMR)) for methyl rotation in the range from 0.4 to 3.2 kcal/mol. The energy differences E(clust) - E(NMR) for each of the ten methyl groups range from -0.2 kcal/mol to +0.7 kcal/mol, with a mean value of +0.2 kcal/mol and a standard deviation of 0.3 kcal/mol. The differences between each of the computed barriers in the clusters (E(clust)) and the corresponding computed barriers in the isolated molecules (E(isol)) provide an estimate of the intermolecular contributions to the rotation barriers in the clusters. The values of E(clust) - E(isol) range from 0.0 to 1.0 kcal/mol. PMID- 21627172 TI - Mechanism of immunoglobulin G4 Fab-arm exchange. AB - Immunoglobulin G (IgG) antibodies are symmetrical molecules that may be regarded as covalent dimers of 2 half-molecules, each consisting of a light chain and a heavy chain. Human IgG4 is an unusually dynamic antibody, with half-molecule exchange ("Fab-arm exchange") resulting in asymmetrical, bispecific antibodies with two different antigen binding sites, which contributes to its anti inflammatory activity. The mechanism of this process is unknown. To elucidate the elementary steps of this intermolecular antibody rearrangement, we developed a quantitative real-time FRET assay to monitor the kinetics of this process. We found that an intrinsic barrier is the relatively slow dissociation of the CH3 domains that noncovalently connect the heavy chains, which becomes rate determining in case disulfide bonds between the heavy chains are reduced or absent. Under redox conditions that mimic the previously estimated in vivo reaction rate, i.e., 1 mM of reduced glutathione, the overall rate is ca. 20 times lower because only a fraction of noncovalent isomers is present (with intra rather than interheavy chain disulfide bonds), formed in a relatively fast pre equilibrium from covalent isomers. Interestingly, Fab arms stabilize the covalent isomer: the amount of noncovalent isomers is ca. 3 times higher for Fc fragments of IgG4 (lacking Fab domains) compared to intact IgG4, and the observed rate of exchange is 3 times higher accordingly. Thus, kinetic data obtained from a sensitive and quantitative real-time FRET assay as described here yield accurate data about interdomain interactions such as those between Fab and/or Fc domains. The results imply that in vivo, the reaction is under control of local redox conditions. PMID- 21627173 TI - Spatiotemporal multicolor labeling of individual cells using peptide functionalized quantum dots and mixed delivery techniques. AB - Multicolor fluorescent labeling of both intra- and extracellular structures is a powerful technique for simultaneous monitoring of multiple complex biochemical processes. This approach remains extremely challenging, however, as it often necessitates the combinatorial use of numerous targeting probes (e.g., antibodies), multistep bioconjugation chemistries, different delivery strategies (e.g., electroporation or transfection reagents), cellular fixation coupled with membrane permeabilization, and complex spectral deconvolution. Here, we present a nanoparticle-based fluorescence labeling strategy for the multicolor labeling of distinct subcellular compartments within live cells without the need for antibody conjugation or cellular fixation/permeabilization. This multipronged approach incorporates an array of delivery strategies, which localize semiconductor quantum dots (QDs) to various subcellular structures. QD uptake is implemented in a spaciotemporal manner by staggering the delivery of QD-peptide composites and exploiting various innate (peptide-mediated endocytosis, peptide-membrane interaction, polymer-based transfection) along with physical (microinjection) cellular delivery modalities to live cells growing in culture over a 4 day period. Imaging of the different intracellular labels is simplified by the unique photophysical characteristics of the QDs in combination with Forster resonance energy transfer sensitization, which allow for multiple spectral windows to be accessed with one excitation wavelength. Using this overall approach, QDs were targeted to both early and late endosomes, the cellular cytosol, and the plasma membrane in live cells, ultimately allowing for simultaneous five-color fluorescent imaging. PMID- 21627297 TI - Controlling reversible elastic deformation of carbon nanotube rings. AB - We show that bundles of carbon nanotubes can be coiled into ring structures by controlling the contraction of their polymer shells. With the robust carbon nanotubes, we demonstrate their reversible transformation between circular and compressed rings in a colloid. PMID- 21627298 TI - Helical superstructure and charged polarons contributions to optical nonlinearity of 2-methyl-4-nitroaniline crystals studied by resonance Raman, electron paramagnetic resonance, circular dichroism spectroscopies, and quantum chemical calculations. AB - The Raman excitation profiles of solid 2-methyl-4-nitroaniline (MNA) reveal several band enhancements by intermolecular and intramolecular charge transfer states. Calculated excited- and ground-state molecular geometries and excited state distortions qualitatively determined from Raman spectra indicate multiple vibrations leading to MNA dissociation. Also, overtones and combination tones can generate charged polarons, as detected by electron paramagnetic resonance after the exposure to 980 and 1550 nm laser diodes. The MNA space group Ia (C(s)(4)) is nonchiral; however, the electronic circular dichroism (CD) spectra of solution, KBr pellet, and single crystal were recorded. The crystal chirality is elucidated by room-temperature dynamic disorder, possible helical superstructure along the [102] polar axis, and charged polarons presence. The CD spectra ab initio calculations for MNA neutral and negatively charged monomers, dimers, and trimers, lying along the helix, confirmed the chirality. The role of these findings toward efficient optical nonlinearity and electric conductivity failure is discussed. PMID- 21627299 TI - Clusters of imidazolium-based ionic liquid in benzene solutions. AB - Cluster formation of 1-dodecyl-3-methylimidazolium bis(trifluoromethanesulfonyl)amide (C(12)mim(+)TFSA(-)) in benzene solutions was investigated using small-angle neutron scattering (SANS), NMR, attenuated total reflectance infrared (ATR-IR), and large-angle X-ray scattering (LAXS) techniques. The SANS measurements revealed that C(12)mim(+)TFSA(-) is heterogeneously mixed with benzene in the narrow range of benzene mole fraction 0.9 <= x(C6D6) <= 0.995 with a maximum heterogeneity at x(C6D6) ~ 0.99. The NMR results suggested that the imidazolium ring is sandwiched between benzene molecules through the cation-pi interaction. Moreover, TFSA(-) probably interacts with the imidazolium ring even in the range of x(C6H6) >= 0.9. Thus, the imidazolium rings, benzene molecules, and TFSA(-) would form clusters in the C(12)mim(+)TFSA(-)-benzene solutions. The LAXS measurements showed that the distance between the imidazolium ring and benzene is ~3.8 A with that between the benzene molecules of ~7.5 A. On the basis of these results, we discussed a plausible reason for the liquid-liquid equilibrium of the C(12)mim(+)TFSA(-) benzene system. PMID- 21627300 TI - Disposable planar reference electrode based on carbon nanotubes and polyacrylate membrane. AB - In this technical note, we report a new all-solid-state planar reference electrode based on single-walled carbon nanotubes and photocured poly(n butylacrylate) (poly(nBA)) membrane containing the Ag/AgCl/Cl(-) ion system. Single-walled carbon nanotubes functionalized with octadecylamide (SWCNT-ODA) and deposited by drop-casting onto a disposable screen-printed electrode are an excellent all-solid-state transducer. The novel potentiometric planar reference electrode shows low potential variability (calibration slopes inferior to 2 mV/dec) for a wide range of chemical species (i.e., ions, small molecules, proteins) in a wide calibration range, redox pairs, changes in pH, and changes in ambient light. Potentiometric medium-term signal stability (-0.9 +/- 0.2 mV/h) and electrochemical impedance characterization confirm the correct solid contact between the SWCNT-ODA layer and photocured poly(nBA) membrane. Overall, the materials used and the simple fabrication by screen-printing and drop-casting enable a high throughput and highly parallel and cost-effective mass manufacture of the new disposable reference electrode. Moreover, the reference electrode has a long shelf life, a characteristic that can be of special interest in decentralized and multiplexing potentiometric analysis. PMID- 21627301 TI - Physicochemical conjugation with deoxycholic acid and dimethylsulfoxide for heparin oral delivery. AB - Heparin, as therapeutic medications, cannot be administered orally because of its hydrophilic and high molecular weight. Here, we present a new technology to enhance the absorption of heparin in the intestine through its chemical conjugation with deoxycholic acid (DOCA) that can interact with bile acid transporter in the intestine. For the ampiphilic property and complete dissolution, the modified heparin was physically complexed with dimethylsulfoxide (DMSO). The DOCA-conjugated heparin could form nanoparticles in aqueous solution, whereas it was completely dissolved when treated with above 10% DMSO solution. Molecular dynamics computation study and two-dimensional homonulcear (1)H nuclear overhauser effect spectroscopy (NOESY) NMR spectra demonstrated that one heparin molecule was chemically conjugated with two DOCA molecules that were physically interacted with six DMSO molecules within 4 A via hydrophobic interactions and partly via hydrogen bonding. Its therapeutic efficacy was also pharmaceutically analyzed. When the DMSO-bound DOCA-conjugated heparin was orally administered into mice, its therapeutic efficacy was enhanced according to the amount of bound DMSO. Also, after oral administration of fluorescence-labeled DMSO-bound DOCA conjugated heparin, it was circulated in the whole body for above 2 h. However, the DOCA-conjugated heparin without DMSO binding was fast eliminated after oral absorption. This study demonstrates that the interaction of structural constraints, DOCA and DMSO, with heparin can serve as a platform technology for potential macromolecule oral delivery. PMID- 21627302 TI - Emergence of superconductivity in "32522" structure of (Ca3Al2O(5-y))(Fe2Pn2) (Pn = As and P). AB - Using a high-pressure technique, we have successfully synthesized (Ca(3)Al(2)O(5 y))(Fe(2)Pn(2)) (Pn = As and P), the first iron-based superconductors with the perovskite-based "32522" structure to be reported. The transition temperature (T(c)) is 30.2 K for Pn = As and 16.6 K for Pn = P. The emergence of superconductivity is ascribed to the small tetragonal a-axis lattice constant of the materials. From these results, an empirical relationship is established between the a-axis lattice constant and T(c) in iron-based superconductors, which offers a practical guideline for exploring new superconductors with higher T(c). PMID- 21627303 TI - Wrinkled nanoporous gold films with ultrahigh surface-enhanced Raman scattering enhancement. AB - Amplified by plasomonic nanostructured metals, Raman intensity of organic molecules and biomolecules can be dramatically improved, particularly at "hot spots" where intense electromagnetic fields are produced in the vicinity of narrow nanogaps between metallic nanostructures. Therefore, developing new substrates with a high density of "hot spots" has been the recent topic of intense study. Here we report wrinkled nanoporous gold films that contain abundant Raman-active nanogaps produced by deformation and fracture of nanowire like gold ligaments. This novel nanostructure yields ultrahigh surface enhanced Raman scattering for molecule detection. PMID- 21627304 TI - Inhibition and activation of glucose oxidase bioanodes for use in a self-powered EDTA sensor. AB - Self-powered sensors are able to automatically signal the presence of a specific analyte without the aid of an external power source, making them useful as potential devices for batteryless sensing. Here, we present a self-powered enzymatic ethylenediaminetetraacetic acid (EDTA) sensor based on the inhibition and subsequent activation of glucose oxidase (GOx)-based bioelectrodes within the framework of a biofuel cell. Although EDTA is not redox-active, it is detected by the activation of a Cu(2+)-inhibited GOx bioanode in either a typical amperometric sensor (using a standard three-electrode setup) or in a self-powered sensor where the GOx bioanode is coupled to a platinum cathode. The sensors are able to detect concentrations of EDTA that correspond to the amount of Cu(2+) that is used to inhibit the enzymatic electrode. The self-powered sensor shows a greater than 10-fold increase in power output when it is activated by the presence of EDTA. This represents the first time that a non-redox-active analyte has been detected in a self-powered sensor that turns on in the presence of said analyte. PMID- 21627305 TI - Operationally simple and highly (E)-styrenyl-selective Heck reactions of electronically nonbiased olefins. AB - Simple, mild, and efficient conditions are reported for a Pd(0)-catalyzed Heck reaction that delivers high yields and selectivity for (E)-styrenyl products using electronically nonbiased olefin substrates bearing a range of useful functionality. Preliminary mechanistic studies demonstrate that the sigma donating DMA solvent is crucial for high selectivity. Further studies suggest that the catalyst distinguishes between beta-hydrogens on the basis of their relative hydridic character, in contrast to previously reported Pd(II)-catalyzed oxidative reaction conditions. PMID- 21627306 TI - Novel application of alpha-azido aldehydes in multicomponent reactions: synthesis of triazolo-fused dihydrooxazinones via a Passerini reaction-dipolar cycloaddition strategy. AB - alpha-Azido aldehydes can be employed in Passerini reactions with isocyanides and various propiolic acids to afford the three-component adducts in moderate to good yields. These compounds undergo a straightforward azide-alkyne dipolar cycloaddition to furnish triazolo-fused dihydrooxazinones. PMID- 21627307 TI - Role of the 3(pipi*) state in photolysis of lumisantonin: insight from ab initio studies. AB - The CASSCF and CASPT2 methodologies have been used to explore the potential energy surfaces of lumisantonin in the ground and low-lying triplet states along the photoisomerization pathways. Calculations indicate that the (1)(npi*) state is the accessible low-lying singlet state with a notable oscillator strength under an excitation wavelength of 320 nm and that it can effectively decay to the (3)(pipi*) state through intersystem crossing in the region of minimum surface crossings with a notable spin-orbital coupling constant. The (3)(pipi*) state, derived from the promotion of an electron from the pi-type orbital mixed with the sigma orbital localized on the C-C bond in the three-membered alkyl ring to the pi* orbital of conjugation carbon atoms, plays a critical role in C-C bond cleavage. Based on the different C-C bond rupture patterns, the reaction pathways can be divided into paths A and B. Photolysis along path A arising from C1-C5 bond rupture is favorable because of the dynamic and thermodynamic preferences on the triplet excited-state PES. Path B is derived from the cleavage of the C5-C6 bond, leading first to a relatively stable species, compared to intermediate A INT formed on the ground state PES. Accordingly, path B is relatively facile for the pyrolytic reaction. The present results provide a basis to interpret the experimental observations. PMID- 21627308 TI - Combinatorial investigation of the effects of the incorporation of Ti, Si, and Al on the performance of alpha-Fe2O3 photoanodes. AB - The effect of adding small amounts of Ti, Si, and Al on the photoelectrochemical activity of alpha-Fe(2)O(3) is investigated using a high-throughput combinatorial method. Quantitative ink jet printing is used to pattern iron oxide and dopant precursors onto conductive glass substrates. Subsequent pyrolysis yields a library of doped iron oxide electrodes that are screened for photoelectrolysis activity by immersing in an electrolyte and scanning a laser over the electrodes to map the photocurrent response. When Si and Al are individually added to iron oxide at the levels we studied, the photoelectrolysis activity decreased whereas low levels of Ti addition enhanced the photocurrents. Synergistic effects were observed resulting in enhanced photocurrents when multiple impurities were added to alpha-Fe(2)O(3). PMID- 21627309 TI - Hydrophobic nanocellulose aerogels as floating, sustainable, reusable, and recyclable oil absorbents. AB - Highly porous nanocellulose aerogels can be prepared by vacuum freeze-drying from microfibrillated cellulose hydrogels. Here we show that by functionalizing the native cellulose nanofibrils of the aerogel with a hydrophobic but oleophilic coating, such as titanium dioxide, a selectively oil-absorbing material capable of floating on water is achieved. Because of the low density and the ability to absorb nonpolar liquids and oils up to nearly all of its initial volume, the surface modified aerogels allow to collect organic contaminants from the water surface. The materials can be reused after washing, recycled, or incinerated with the absorbed oil. The cellulose is renewable and titanium dioxide is not environmentally hazardous, thus promoting potential in environmental applications. PMID- 21627310 TI - Organoplatinum chromophores for application in high-performance nonlinear absorption materials. AB - Chromophores and materials that exhibit nonlinear absorption over a broad spectrum and with high temporal dynamic range are of interest for application in materials engineering and biology. Recent work by a number of research groups has led to the development of a new family of organometallic chromophores and materials featuring interesting and useful nonlinear absorption properties. These systems contain the platinum acetylide moiety as a fundamental molecular unit, combined with delocalized, pi-conjugated electron systems. These organometallic chromophores provide a unique combination of properties, such as negligible ground state absorption in the visible region, large spin-orbit coupling giving rise to high triplet excited state yield, triplet lifetime in the microsecond domain, high two-photon cross-section in the visible and near-infrared regions, and high triplet-triplet absorption cross-section in the visible and near infrared region. This Spotlight on Application highlights recent developments in this area, combining background and review on nonlinear absorption in platinum acetylide chromophores and describing significant recent results from our own laboratory. PMID- 21627311 TI - Experimental study of the quantitative precision for valve-based comprehensive two-dimensional gas chromatography. AB - For complex sample analysis, there is a need for multidimensional chromatographic instrumentation to be able to separate more compounds, often in shorter time frames. This has led to the development of comprehensive two-dimensional chromatographic instrumentation, such as comprehensive two-dimensional gas chromatography (GC * GC). Lately, much of the focus in this field has been on decreasing peak widths and, therefore, increasing peak capacity and peak capacity production. All of these advancements make it possible to analyze more compounds in a shorter amount of time, but the data still need to remain quantitative to address the needs of most applications. In this report, the relationship among the modulation ratio (M(R)), peak sampling phase (phi), retention time variation (Deltat(R)), and how these parameters relate to quantitative analysis precision via the relative standard deviation (RSD) was studied experimentally using a valve-based GC * GC instrument. A wide range of the number of modulations across the first dimension peak width, that is, a M(R) range from ~1 to 10, was examined through maintaining an average first dimension peak width at the base, (1)w(b) of ~3 s and varying the second dimension separation run time from 300 to 2900 ms. An average RSD of 2.1% was experimentally observed at an average M(R) of 2, with a corresponding peak capacity production of ~1200 peaks/min possible. Below this M(R) the RSD quickly increased. In a long-term study of the quantitative precision at a M(R) of 2.5, using 126 replicate injections of a test mixture spanning ~35 h, the RSD averaged 3.0%. The findings have significant implications for optimizing peak capacity production by allowing the use of the longest second dimension run time, while maintaining quantitative precision. PMID- 21627312 TI - General synthesis of dinaphtho[2,3-b:2',3'-f]thieno[3,2-b]thiophene (DNTT) derivatives. AB - A new straightforward synthesis of dinaphtho[2,3-b:2',3'-f]thieno[3,2-b]thiophene (DNTT) derivatives from readily available 2-methoxynaphthalenes is described. Thus, newly developed derivatives of DNTT showed very high field effect mobility in the vapor-processed field-effect transistors up to 8 cm(2) V(-1) s(-1). PMID- 21627313 TI - Mechanistic insights on N-heterocyclic carbene-catalyzed annulations: the role of base-assisted proton transfers. AB - The density functional theory investigation on the mechanism of NHC-catalyzed cycloannulation reaction of the homoenolate derived from butenal with pentenone is studied. The M06-2X/6-31+G** and B3LYP/6-31+G** levels of theory, including the effect of continuum solvation in dichloromethane and tetrahydrofuran, are employed. Several mechanistic scenarios are examined for each elementary step by identifying the key intermediates and the corresponding transition states interconnecting them on the respective potential energy surfaces. Both assisted and unassisted pathways for important proton transfer steps are considered, respectively, with and without the explicit inclusion of base (DBU) in the corresponding transition states. The barrier for the crucial proton transfer steps involved in the formation of the Breslow intermediate as well as in the subsequent steps is found to be significantly lowered by explicit inclusion of DBU. The energetic comparison between two key pathways, depicted as path A and path B, respectively, leading to cyclopentene and cyclopentanone derivatives, is performed. The major mechanistic bifurcation has been identified as emanating from the site of enolization of the initial zwitterionic intermediate resulting from the addition of a homoenolate equivalent to enone. If the enolization occurs nearer to the NHC moiety, the reaction is likely to proceed through path A, leading to cyclopentene. The enolization away from NHC leads to cyclopentanone product through path B. The computed results are generally in good agreement with the reported experimental results. PMID- 21627314 TI - Electrochemical and spectroscopic studies on the conformational structure of hemoglobin assembled on gold nanoparticles. AB - Protein conformational changes may be associated with particular properties such as function, transportation, assembly, tendency to aggregate, and potential cytotoxicity. Protein misfolding, in particular, has been intimately related to protein-mediated diseases. In this study, the conformational structure changes of hemoglobin (Hb) induced by the assembly on gold nanoparticles (AuNPs) surface were studied in detail by a combination of electrochemical method and various spectroscopic techniques including UV-vis absorption, fluorescence, circular dichroism (CD), and Fourier transform infrared (FTIR) spectroscopy. The results indicated that Hb in the Hb-AuNPs bioconjugate system that was prepared by the assembly of Hb on the surface of AuNPs underwent substantial conformational changes both at secondary and tertiary structure level. The assembly of Hb on the boundary surface of AuNPs could result a disturbance of the structure of Hb and induce the exposure of the heme group and tryptophan (Trp) residues to the solvent, leading to the enhancement in the electron transfer rate of the protein. The calculation from quantitative second-derivative infrared and CD spectra of the Hb-AuNPs bioconjugate system showed that AuNPs could induce the conversion of alpha-helix to beta-sheet structures and unfolding of the protein. Moreover, the effects of the concentration and the size of AuNPs on the conformational structure changes of Hb in the bioconjugate system were also demonstrated. The results obtained here not only provide the detailed conformational behavior of Hb molecules on nanoparticles, but also create a framework for analyzing the biosafety of nanoparticles in terms of the biological behavior of biomacromolecules. PMID- 21627315 TI - Maxwell-Stefan diffusivities in binary mixtures of ionic liquids with dimethyl sulfoxide (DMSO) and H2O. AB - Ionic liquids (ILs) are promising solvents for applications ranging from CO2 capture to the pretreatment of biomass. However, slow diffusion often restricts their applicability. A thorough understanding of diffusion in ILs is therefore highly desirable. Previous research largely focused on self-diffusion in ILs. For practical applications, mutual diffusion is by far more important than self diffusion. For describing mutual diffusion in multicomponent systems, the Maxwell Stefan (MS) approach is commonly used. Unfortunately, it is difficult to obtain MS diffusivities from experiments, but they can be directly extracted from molecular dynamics (MD) simulations. In this work, MS diffusivities were computed in binary systems containing 1-alkyl-3-methylimidazolium chloride (C(n)mimCl, n = 2, 4, 8), water, and/or dimethyl sulfoxide (DMSO) using MD. The dependence of self- and MS diffusivities on mixture composition was investigated. Our results show the following: (1) For solutions of ILs in water and DMSO, self diffusivities decrease strongly with increasing IL concentration. For DMSO-IL, a single exponential decay is observed. (2) In both water-IL and DMSO-IL, MS diffusivities vary by a factor of 10 within the concentration range which is, however, still significantly smaller than the variation of the self-diffusion coefficients. (3) The MS diffusivities of the IL are almost independent of the alkyl chain length. (4) ILs stay in a form of isolated ions in C(n)mimCl-H2O mixtures; however, dissociation into ions is much less observed in C(n)mimCl-DMSO systems. This has a large effect on the concentration dependence of MS diffusivities. (5) Recently, we proposed a new model for predicting the MS diffusivity at infinite dilution, that is, D(ij)(x(k-->)1) (Ind. Eng. Chem. Res. 2011, 50, 4776-4782). This quantity describes the friction between components i and j when both are infinitely diluted in component k. In contrast to earlier empirical models, our model is based on the linear response theory and the Onsager relations which allows a clear interpretation of the results. The key assumption in the model is that velocity cross-correlations are neglected. The present study clearly shows that velocity cross-correlation functions in ILs cannot be neglected and that the dissociation of ILs into ions has a very strong influence on diffusion. PMID- 21627316 TI - Diastereo- and enantioselective ruthenium-catalyzed hydrohydroxyalkylation of 2 silyl-butadienes: carbonyl syn-crotylation from the alcohol oxidation level. AB - Exposure of alcohols 2a-2j to 2-silyl-butadienes in the presence of ruthenium complexes modified by (R)-SEGPHOS or (R)-DM-SEGPHOS results in redox-triggered generation of allylruthenium-aldehyde pairs, which combine to form products of carbonyl crotylation 4a-4j in the absence of stoichiometric byproducts and with high levels of syn-diastereo- and enantioselectivity. In the presence of isopropanol under otherwise identical conditions, aldehydes 3a-3j are converted to an equivalent set of adducts 4a-4j. Whereas reactions conducted using conventional heating require 48 h, microwave irradiation enables full conversion in only 4 h. Finally, as illustrated in the conversion of adduct 4a to compounds 6a and 6b, diastereoselective hydroboration-Suzuki cross-coupling with aryl and vinyl halides followed by Fleming-Tamao oxidation enables generation of anti,syn stereotriads found in numerous polyketide natural products. PMID- 21627317 TI - Hierarchical helical assembly of conjugated poly(3-hexylthiophene)-block-poly(3 triethylene glycol thiophene) diblock copolymers. AB - We report on the solution-state assembly of all-conjugated polythiophene diblock copolymers containing nonpolar (hexyl) and polar (triethylene glycol) side chains. The polar substituents provide a large contrast in solubility, enabling formation of stably suspended crystalline fibrils even under very poor solvent conditions for the poly(3-hexylthiophene) block. For appropriate block ratios, complexation of the triethylene glycol side chains with added potassium ions drives the formation of helical nanowires that further bundle into superhelical structures. PMID- 21627318 TI - Effect of simultaneous consumption of milk and coffee on chlorogenic acids' bioavailability in humans. AB - Different studies have shown that milk may interact with polyphenols and affect their bioavailability in humans. The present study investigated the effect of the simultaneous consumption of coffee and milk on the urinary excretion of chlorogenic acids (CGA) and metabolites. Subjects were submitted to consumption of water, instant coffee (609 mmol of CGA) dissolved in water, and instant coffee dissolved in whole milk. Urine was collected for 24 h after consumption of each treatment for analysis of CGA and metabolites by HPLC/LC-MS. The amount of CGA and metabolites recovered after consumption of combined coffee-milk (40% +/- 27%) was consistently lower in all subjects compared to that of coffee alone (68% +/- 20%). Concluding, the simultaneous consumption of milk and coffee may impair the bioavailability of coffee CGA in humans. PMID- 21627319 TI - Assessment of soy genotype and processing method on quality of soybean tofu. AB - Protein quality in six soybean varieties, based on subunit composition of their protein, was correlated with quality of the produced tofu. Also, protein changes due to a pilot plant processing method involving high temperature/pressure and commercial rennet as coagulant were assessed. In each soybean variety, glycinin (11S) and beta-conglycinin (7S) as well as 11S/7S ratio significantly changed from beans to tofu. Between varieties, the 11S/7S protein ratio in seed indicated genotypic influence on tofu yield and gel hardness (r = 0.91 and r = 0.99, respectively; p < 0.05). Also, the 11S/7S ratio correlated with soymilk pH (r = 0.89, p < 0.05), leading to a relationship between soymilk pH with protein recovery and yield of tofu (r = 0.94 and r = 0.91, respectively; p < 0.05). The soybean beta'-subunit of 7S protein negatively influenced tofu hardness (r = 0.91, p < 0.05). Seed protein composition and proportion of 7S protein subunits under the applied production method had an important role in defining tofu quality. PMID- 21627320 TI - Quantitative colorimetric assay for total protein applied to the red wine Pinot noir. AB - A standard method for assaying protein in red wine is currently lacking. The method described here is based on protein precipitation followed by dye binding quantification. Improvements over existing approaches include minimal sample processing prior to protein precipitation with cold trichloroacetic acid/acetone and quantification based on absorbance relative to a commercially available standard representative of proteins likely to be found in wine, the yeast mannoprotein invertase. The precipitation method shortened preparation time relative to currently published methods and the mannoprotein standard yielded values comparable to those obtained by micro-Kjeldahl analysis. The assay was used to measure protein in 48 Pinot noir wines from 6 to 32 years old. The protein content of these wines was found to range from 50 to 102 mg/L with a mean value of 70 mg/L. The availability of a simple and relatively rapid procedure for assaying protein provides a practical tool to quantify a wine component that has been overlooked in routine analyses of red wines. PMID- 21627321 TI - Quantitative iTRAQ analysis of retinal ganglion cell degeneration after optic nerve crush. AB - Retinal ganglion cells (RGCs) are central nervous system (CNS) neurons that transmit visual information from the retina to the brain. Apoptotic RGC degeneration causes visual impairment that can be modeled by optic nerve crush. Neuronal apoptosis is also a salient feature of CNS trauma, ischemia (stroke), and diseases of the CNS such as Alzheimer's, Parkinson's, multiple sclerosis, and amyotrophic lateral sclerosis. Optic nerve crush induces the apoptotic cell death of ~ 70% of RGCs within the first 14 days after injury. This model is particularly attractive for studying adult neuron apoptosis because the time course of RGC death is well established and axon regeneration within the myelinated optic nerve can be concurrently evaluated. Here, we performed a large scale iTRAQ proteomic study to identify and quantify proteins of the rat retina at 1, 3, 4, 7, 14, and 21 days after optic nerve crush. In total, 337 proteins were identified, and 110 were differentially regulated after injury. Of these, 58 proteins were upregulated (>1.3 *), 46 were downregulated (<0.7 *), and 6 showed both positive and negative regulation over 21 days, relative to normal retinas. Among the differentially expressed proteins, Thymosin-beta4 showed an early upregulation at 3 days, the time-point that immediately precedes the induction of RGC apoptosis after injury. We examined the effect of exogenous Thymosin-beta4 administration on RGC death after optic nerve injury. Intraocular injections of Thymosin-beta4 significantly increased RGC survival by ~ 3-fold compared to controls and enhanced axon regeneration after crush, demonstrating therapeutic potential for CNS insults. Overall, our study identified numerous proteins that are differentially regulated at key time-points after optic nerve crush, and how the temporal profiles of their expression parallel RGC death. This data will aid in the future development of novel therapeutics to promote neuronal survival and regeneration in the adult CNS. PMID- 21627322 TI - Proteomic analyses of lung lysates from short-term exposure of Fischer 344 rats to cigarette smoke. AB - A short-term 5 day mainstream cigarette smoke exposure study was conducted in Fischer 344 rats to identify changes in lung proteins. Groups of 10 male and female rats at 5 weeks of age were assigned to one of four exposure groups. Animals received either nose-only filtered air (Air Control) or 75, 200, or 400 mg total particulate matter (TPM)/m(3) of diluted cigarette smoke. Exposures were conducted for 3 h per day, for 5 consecutive days. One lung per animal was frozen in liquid nitrogen and processed for proteomic analyses. Lung lysates from control verses treated animals were screened with 650 antibodies for changes in signaling protein levels and phosphorylation using antibody microarray technology, and then over 100 of the top protein hits were assessed by immunoblotting. The top smoke-altered proteins were further evaluated using reverse lysate microarrays. Major protein changes showed medium to strong bands on Western blots, depended on dose and gender, and included protein-serine kinases (Cot/Tpl2, ERK1/2, GSK3alpha/beta, MEK6, PKCalpha/gamma, RSK1), protein phosphatases (PP4/A'2, PP1Cbeta), and other proteins (caspase 5, CRMP2, Hsc70, Hsp60, Rac1 and STAT2). The most pronounced changes occurred with 75 mg TPM/m(3) exposed females and 200 mg TPM/m(3) exposed males. Smoke-altered proteins regulate apoptosis, stress response, cell structure, and inflammation. Changes in identified proteins may serve as early indicators of lung damage. PMID- 21627323 TI - Light wavelength effects on a lutein-fortified model colloidal beverage. AB - The effect of light on a model colloidal beverage system containing whey protein, lutein, and limonene was investigated. Changes in volatile chemistry were evaluated under accelerated conditions (12 h, 25 degrees C) at selected wavelengths regions (395, 463, 516, 567, and 610 nm absorbance maxima) using a photochemical reactor. The most damaging wavelengths to lutein stability were UV (200-400 nm) and 463 nm wavelengths. Hexanal formation was highest in the control beverage when exposed to full spectrum light and UV (200-400 nm) wavelengths. Hexanal also was formed in the lutein-fortified beverage under full spectrum light and UV (200-400 nm) wavelengths but to a significantly lesser degree. Limonene degraded significantly under all treatment conditions, with most degradation occurring during full spectrum light exposure. Lutein fortification did not completely protect limonene from degradation. PMID- 21627324 TI - Quality and aromatic sensory descriptors (mainly fresh and dry fruit character) of Spanish red wines can be predicted from their aroma-active chemical composition. AB - A satisfactory model explaining quality could be built in a set of 25 high quality Spanish red wines, by aroma-active chemical composition. The quality of the wines was positively correlated with the wine content in fruity esters, acids, enolones, and wood derived compounds, and negatively with phenylacetaldehyde, acetic acid, methional, and 4-ethylphenol. Wine fruitiness was demonstrated to be positively related not only to the wine content on fruity esters and enolones, but to wine volatile fatty acids. Fruitiness is strongly suppressed by 4-ethylphenol, acetic acid, phenylacetaldehyde, and methional, this involved in the perception of dry-fruit notes. Sensory effects were more intense in the presence of beta-damascenone and beta-ionone. A satisfactory model explaining animal notes could be built. Finally, the vegetal character of this set of wines could be related to the combined effect of dimethylsulfide (DMS), 1 hexanol, and methanethiol. PMID- 21627325 TI - Lunasin and Bowman-Birk protease inhibitor concentrations of protein extracts from enzyme-assisted aqueous extraction of soybeans. AB - Lunasin and Bowman-Birk protease inhibitor (BBI) are two soybean peptides to which health-promoting properties have been attributed. Concentrations of these peptides were determined in skim fractions produced by enzyme-assisted aqueous extraction processing (EAEP) of extruded full-fat soybean flakes (an alternative to extracting oil from soybeans with hexane) and compared with similar extracts from hexane-defatted soybean meal. Oil and protein were extracted by using countercurrent two-stage EAEP of soybeans at 1:6 solids-to-liquid ratio, 50 degrees C, pH 9.0, and 120 rpm for 1 h. Protein-rich skim fractions were produced from extruded full-fat soybean flakes using different enzyme strategies in EAEP: 0.5% protease (wt/g extruded flakes) used in both extraction stages; 0.5% protease used only in the second extraction stage; no enzyme used in either extraction stage. Countercurrent two-stage protein extraction of air desolventized, hexane-defatted soybean flakes was used as a control. Protein extraction yields increased from 66% to 89-96% when using countercurrent two stage EAEP with extruded full-fat flakes compared to 85% when using countercurrent two-stage protein extraction of air-desolventized, hexane-defatted soybean flakes. Extruding full-fat soybean flakes reduced BBI activity. Enzymatic hydrolysis reduced BBI contents of EAEP skims. Lunasin, however, was more resistant to both enzymatic hydrolysis and heat denaturation. Although using enzymes in both EAEP extraction stages yielded the highest protein and oil extractions, reducing enzyme use to only the second stage preserved much of the BBI and Lunasin. PMID- 21627326 TI - Remarkable metal-complexed phosphorus analogues of the cyclopropenylcarbene cyclobutadiene rearrangement. AB - In situ-generated metal carbonyl-complexed cyclopropenylphosphinidenes undergo a sequence of structural changes leading to phosphorus analogues of Pettit's seminal (eta(4)-cyclobutadiene)iron tricarbonyl complex via multiple valence isomers along the reaction pathway and the elimination of one molecule of carbon monoxide. PMID- 21627327 TI - Structure-based fragment hopping for lead optimization using predocked fragment database. AB - In this work, we describe a structure-based de novo optimization process, called "LeadOp" (short for lead optimization), that decomposes a compound into fragments of different molecular components either by chemical or user-defined rules. Each fragment is evaluated through a predocked fragment database that ranks fragments according to specific fragment-receptor binding interactions, replacing fragments that contribution the least to binding and finally reassembling the fragments to form a new ligand. The fundamental idea is to replace "bad" fragments of a ligand with "good" fragments while leaving the core of the ligand intact, thus improving the compound's activity. The molecular fragments were selected from a collection of 27,417 conformers that are the fragments of compounds in the DrugBank database. The collection of molecular fragments are docked to the target's binding site and evaluated using group efficiency (calculated binding affinity divided by the number of heavy atoms), and the "strongest" binder is selected. The LeadOp method was tested with two biomolecular systems: mutant B-Raf kinase and human 5-lipoxygenase. The LeadOp methodology was able to optimize the query molecules and systematically developed improved analogs for each of our example systems. PMID- 21627328 TI - Synthesis, analytical features, and biological relevance of 5-(3',4' dihydroxyphenyl)-gamma-valerolactone, a microbial metabolite derived from the catabolism of dietary flavan-3-ols. AB - The physiological significance of 5-(3',4'-dihydroxyphenyl)-gamma-valerolactone, an important metabolite derived from the catabolism of flavan-3-ols by gut microbiota, has been often overlooked due to the lack of the commercial standard. In the present work, this metabolite has been chemically synthesized, and its analytical parameters and antioxidant capacity have been determined in comparison to other chemical analogues [isomer 3-(3',4'-dihydroxyphenyl)-delta-valerolactone and gamma-valerolactone] and other structurally related compounds [(+)-catechin, (-)-epicatechin, and 3-(3,4-dihydroxyphenyl)-propionic acid]. The synthesized compound was also used to perform a targeted analysis in samples collected during the in vitro fermentation of a grape seed flavan-3-ol extract with human fecal microbiota from three healthy volunteers. The time-course formation of 5-(3',4' dihydroxyphenyl)-gamma-valerolactone revealed large interindividual differences among volunteers, with concentrations ranging from 3.31 to 77.54 MUM at 10 h of fermentation. These results are further discussed in view of the scarce reports quantifying 5-(3',4'-dihydroxyphenyl)-gamma-valerolactone in in vitro fermentation studies, and pharmacokinetic and intervention studies. PMID- 21627329 TI - Iron-promoted C-C bond cleavage of 1,3-diketones: a route to 1,2-diketones under mild reaction conditions. AB - A conceptual method for the preparation of 1,2-diketones is reported. The selective C-C bond cleavage of 1,3-diketones affords the 1,2-diketones in high yields under mild reaction conditions in air by the use of FeCl(3) as the catalyst and tert-butyl nitrite (TBN) as the oxidant without the use of solvent. The possible reaction mechanism is discussed. This protocol provides an expeditious route to the useful 1,2-diketones. PMID- 21627330 TI - Molecular mechanism of action of beta-hairpin antimicrobial peptide arenicin: oligomeric structure in dodecylphosphocholine micelles and pore formation in planar lipid bilayers. AB - The membrane-active, cationic, beta-hairpin peptide, arenicin, isolated from marine polychaeta Arenicola marina exhibits a broad spectrum of antimicrobial activity. The peptide in aqueous solution adopts the significantly twisted beta hairpin conformation without pronounced amphipathicity. To assess the mechanism of arenicin action, the spatial structure and backbone dynamics of the peptide in membrane-mimicking media and its pore-forming activity in planar lipid bilayers were studied. The spatial structure of the asymmetric arenicin dimer stabilized by parallel association of N-terminal strands of two beta-hairpins was determined using triple-resonance nuclear magnetic resonance (NMR) spectroscopy in dodecylphosphocholine (DPC) micelles. Interaction of arenicin with micelles and its oligomerization significantly decreased the right-handed twist of the beta hairpin, increased its amphipathicity, and led to stabilization of the peptide backbone on a picosecond to nanosecond time scale. Relaxation enhancement induced by water-soluble (Mn(2+)) and lipid-soluble (16-doxylstearate) paramagnetic probes pointed to the dimer transmembrane arrangement. Qualitative NMR and circular dichroism study of arenicin-2 in mixed DPC/1,2-dioleoyl-sn-glycero-3 phosphoglycerol bicelles, sodium dodecyl sulfate micelles, and lipid vesicles confirmed that a similar dimeric assembly of the peptide was retained in membrane mimicking systems containing negatively charged lipids and detergents. Arenicin induced conductance was dependent on the lipid composition of the membrane. Arenicin low-conductivity pores were detected in the phosphatidylethanolamine containing lipid mixture, whereas the high-conductivity pores were observed in an exclusively anionic lipid system. The measured conductivity levels agreed with the model in which arenicin antimicrobial activity was mediated by the formation of toroidal pores assembled of two, three, or four beta-structural peptide dimers and lipid molecules. The structural transitions involved in arenicin membrane disruptive action are discussed. PMID- 21627331 TI - Self-assembled functional organic monolayers on oxide-free copper. AB - The preparation and characterization of self-assembled monolayers on copper with n-alkyl and functional thiols was investigated. Well-ordered monolayers were obtained, while the copper remained oxide-free. Direct attachment of N succinimidyl mercaptoundecanoate (NHS-MUA) onto the copper surface allowed for the successful attachment of biomolecules, such as beta-d-glucosamine, the tripeptide glutathione, and biotin. Notably, the copper surfaces remained oxide free even after two reaction steps. All monolayers were characterized by static water contact angle measurements, X-ray photoelectron spectroscopy, and infrared reflection absorption spectroscopy. In addition, the biotinylated copper surfaces were employed in the immobilization of biomolecules such as streptavidin. PMID- 21627332 TI - The conformational flexibility of the helicase-like domain from Thermotoga maritima reverse gyrase is restricted by the topoisomerase domain. AB - Reverse gyrase is the only enzyme known to introduce positive supercoils into DNA. Positive supercoiling is achieved by the functional cooperation of a helicase-like and a topoisomerase domain. The isolated helicase-like domain is a DNA-stimulated ATPase, and the isolated topoisomerase domain can relax supercoiled DNA. In the context of reverse gyrase, these individual activities are suppressed or attenuated. The helicase-like domain of Thermotoga maritima reverse gyrase is a nucleotide-dependent conformational switch that binds DNA and ATP cooperatively. It provides a nucleotide-dependent DNA-binding site to reverse gyrase and thus serves as a valuable model for the investigation of the effect of nucleotides on DNA processing by reverse gyrase that is key to its supercoiling activity. To improve our understanding of the structural basis for the functional cooperation of a helicase domain with a DNA topoisomerase, we have determined the structures of the isolated helicase-like domain of T. maritima reverse gyrase in five different conformations. Comparison of these structures reveals extensive domain flexibility in the absence of conformational restrictions by the topoisomerase that is consistent with single-molecule Forster resonance energy transfer experiments presented here. The structure of the first ADP-bound form provides novel details about nucleotide binding to reverse gyrase. It demonstrates that reverse gyrases use the canonical nucleotide binding mode common to superfamily 2 helicases despite large deviations in the conserved motifs. A characteristic insert region adopts drastically different structures in different reverse gyrases. Counterparts of this insert region are located at very different positions in other DNA-processing enzymes but may point toward a general role in DNA strand separation. PMID- 21627333 TI - Sesquarterpenes (C35 terpenes) biosynthesized via the cyclization of a linear C35 isoprenoid by a tetraprenyl-beta-curcumene synthase and a tetraprenyl-beta curcumene cyclase: identification of a new terpene cyclase. AB - In this study, mono- and pentacyclic C(35) terpenes from Bacillus subtilis were biosynthesized via the cyclization of C(35) isoprenoid using purified enzymes, including the first identified new terpene cyclase that shows no sequence homology to any of the known terpene cyclases. On the basis of these findings, we propose that these C(35) terpenes should be called the new family of "sesquarterpenes." PMID- 21627335 TI - Administration of CREON(r) pancrelipase pellets via gastrostomy tube is feasible with no loss of gastric resistance or lipase activity: an in vitro study. AB - BACKGROUND AND OBJECTIVES: In clinical practice, the need sometimes arises to administer pancreatic enzyme replacement therapy via gastrostomy tube (G-tube) by mixing the pellets contained in the capsules with soft food. The objective of this study was to identify G-tubes that allow administration of pancrelipase gastro-resistant pellets without clogging, sticking, pellet damage or loss of enteric coating integrity. METHODS: In this in vitro study, CREON(r) (pancrelipase) Delayed-Release Capsules were opened and the pellets sprinkled onto a small amount of baby food of pH <4.5 (applesauce and bananas manufactured by both Gerber and Beech-Nut). The mixture was stirred gently and after 15 minutes poured into a 35 mL syringe and pushed slowly (~15 mL in 10-15 seconds) through a G-tube. Pellets were collected and the tube flushed with water. G-tubes were inspected visually for clogging/sticking and damage to pellets was assessed. If there was none with all four foods, pellet integrity (gastric resistance and lipase activity) was assessed by an in vitro dissolution method with a 2-hour gastric simulation step. The activity required to confirm integrity was >=80% of actual US Pharmacopeia lipase activity per capsule. G-tubes initially tested were Kimberly-Clark MIC Bolus(r) size 14 French (Fr) and upwards and Kimberly-Clark MIC-KEY(r) 14 Fr and upwards. Following successful testing, assessment of Bard(r) Tri-Funnel 18 Fr and Bard(r) Button 18 Fr G-tubes was carried out. RESULTS: Based on the absence of clogging, sticking and visible damage to pellets, and the maintenance of pellet integrity, administration of CREON(r) pancrelipase pellets was feasible through the following G-tubes: Kimberly-Clark MIC Bolus(r) size 18 Fr, Kimberly-Clark MIC-KEY(r) 16 Fr, Bard(r) Tri-Funnel 18 Fr and Bard(r) Button 18 Fr. Lipase activity met the predetermined specification and was >=90% for all four tubes and all four foods, with no differences versus untreated pellets (i.e. pellets not mixed with baby food or pushed through a G-tube). These data apply to all CREON(r) pancrelipase capsule formulations, regardless of their strength in lipase units, as pellet composition, size and quality are identical. CONCLUSION: CREON(r) pancrelipase pellets can be mixed with baby food of pH <4.5 and administered via the following G-tubes without clogging, sticking or visible pellet damage, and with no loss of gastric resistance or lipase activity: Kimberly-Clark MIC Bolus(r) size 18 Fr and larger, Kimberly-Clark MIC-KEY(r) 16 Fr and larger, Bard(r) Tri-Funnel 18 Fr and larger and Bard(r) Button 18 Fr and larger. PMID- 21627336 TI - Combination delapril/manidipine as antihypertensive therapy in high-risk patients. AB - The majority of patients with hypertension, and in particular high-risk patients or those with diabetes mellitus or renal dysfunction, are likely to require combination therapy with at least two antihypertensive agents (from different classes) to achieve their blood pressure (BP) target. The delapril/manidipine fixed-dose combination consists of two antihypertensive agents with different, yet complementary, mechanisms of action. Delapril/manidipine has demonstrated short- and long-term antihypertensive efficacy in a number of clinical studies in patients with hypertension with an inadequate response to monotherapy. Comparative studies have demonstrated that delapril/manidipine is as effective as enalapril/hydrochlorothiazide (HCTZ) in patients with hypertension with an inadequate response to monotherapy, and as effective as irbesartan/HCTZ, losartan/HCTZ, olmesartan medoxomil/HCTZ, ramipril/HCTZ and valsartan/HCTZ in reducing BP in patients with hypertension and diabetes, or in obese patients with hypertension. Therapy with delapril/manidipine also appears to exert beneficial effects that extend beyond a reduction in BP, including nephroprotective activity and an improvement in fibrinolytic balance, supporting its value as a treatment option in these patient populations at high or very high cardiovascular risk because of the presence of organ damage, diabetes or renal disease. PMID- 21627337 TI - Identifying amnestic mild cognitive impairment in primary care: a feasibility study. AB - BACKGROUND AND OBJECTIVE: Amnestic mild cognitive impairment (aMCI), characterized by episodic memory impairment in the absence of clinical dementia, often represents a transitional stage between normal aging and Alzheimer's disease (AD). It is not known if non-expert primary-care physicians (PCPs) can differentiate individuals with no cognitive impairment (NCI), aMCI and mild AD in a primary-care practice setting. This study develops an approach to this question, which is necessary for aMCI to become a treatment target. METHODS: Fourteen experts assessed subjects with memory complaints in terms of their laboratory test results, magnetic resonance imaging findings and scores on the Mini-Mental State Examination, adapted Clinical Dementia Rating Scale and Alzheimer's Disease Assessment Scale-cognitive subscale Delayed Word Recall before designating each subject as having NCI, aMCI or AD. Subjects agreed upon by a consensus committee were assigned to non-expert PCPs who, following brief training, assessed them using the same clinical information and utilizing the same assessment instruments. The chance-corrected inter-rater reliability (expert versus non-expert) measure kappa, based on binary outcome (aMCI/not-aMCI), was estimated. RESULTS: The study recruited 119 evaluable subjects (50 aMCI, 27 mild AD and 42 NCI) and demonstrated fair to moderate agreement (kappa = 0.423) between experts and non-experts in designation of aMCI. The percent agreement was 72.3%, sensitivity 62.0% and specificity 79.7%. Overall, non-experts under-rated the level of impairment compared with experts. CONCLUSION: This study established the feasibility of making the aMCI designation in the community and identified some likely sources of error. The results suggest that when drugs with clear benefit for aMCI patients are developed, community-based PCPs, with additional, more optimized training, will be able to accurately identify those patients who should receive treatment. PMID- 21627338 TI - Iron-chelating therapies in a transfusion-dependent thalassaemia population in Thailand: a cost-effectiveness study. AB - BACKGROUND AND OBJECTIVE: beta-Thalassaemia is a major public health problem in Thailand. Use of appropriate iron-chelating agents could prevent thalassaemia related complications, which are costly to the healthcare system. This study aimed to evaluate the cost effectiveness of deferoxamine (DFO), deferiprone (DFP) and deferasirox (DFX) in Thai transfusion-dependent beta-thalassaemia patients from the societal perspective. METHODS: A Markov model was used to project the life-time costs and outcomes represented as quality-adjusted life-years (QALYs). Data on the clinical efficacy and safety of all therapeutic options were obtained from a systematic review and clinical trials. Transition probabilities were derived from published studies. Costs were obtained from the Thai Drug and Medical Supply Information Center, Thai national reimbursement rate information and other Thai literature sources. A discount rate of 3% was used. Incremental cost-effectiveness ratios (ICERs) were presented as year 2009 values. A base-case analysis was performed for thalassaemia patients requiring regular blood transfusion therapy, while a separate analysis was performed for patients requiring low (i.e. symptom-dependent, less frequent) blood transfusion therapy. A series of sensitivity analysis and cost-effectiveness acceptability curves were constructed. RESULTS: Compared with DFO, using DFP was dominant with lifetime cost savings of $US91 117. Comparing DFX with DFO, the incremental cost was $US522 863 and incremental QALY was 5.77 with an ICER of $US90 648 per QALY. When compared with DFP, the ICER of DFX was $US106 445 per QALY. A cost-effectiveness analysis curve showed the probability of DFX being cost effective was 0% when compared with either DFO or DFP, based on the cost-effectiveness cut-off value of $US2902 per QALY. When compared with DFP, DFX was cost effective only if the DFX cost was as low as $US1.68 per 250 mg tablet. The results of the analysis in patients requiring low blood transfusion therapy were not different from those of the base-case analysis. CONCLUSIONS: Our findings suggest that using DFP is cost saving when compared with conventional therapy, while using DFX is not cost effective compared with either DFO or DFP in Thai patients with transfusion dependent beta-thalassaemia. Policy-makers and clinicians may consider using such information in their decision-making process in Thailand. PMID- 21627339 TI - Bevacizumab plus interferon-alpha versus sunitinib for first-line treatment of renal cell carcinoma in Italy: a cost-minimization analysis. AB - BACKGROUND: Renal cell carcinoma (RCC) is the most common form of kidney cancer. Immunotherapy with interferon-alpha (IFNalpha) and interleukin-2 (IL-2) has been the historical therapy of choice for the treatment of locally advanced or metastatic RCC prior to the more recent development of targeted therapies, including sunitinib and bevacizumab (combined with IFNalpha). Clinically and statistically significant advantages have been shown with both sunitinib and the combination of bevacizumab + IFNalpha versus IFNalpha alone in the treatment of advanced or metastatic RCC. The present study evaluated the incremental costs of bevacizumab + IFNalpha versus sunitinib for the first-line treatment of advanced or metastatic RCC assuming similar efficacy for these treatments. METHODS: The efficacy profiles of bevacizumab + IFNalpha or sunitinib alone have been shown (indirectly) to be similar in patients with RCC; indeed, median progression-free survival (PFS) with either treatment is in the 10- to 11-month range. Therefore, a cost-minimization analysis was performed, focusing on direct medical costs only (drugs, administration and management of adverse events). The analysis considered the perspective of the Italian National Health Service (NHS), comparing the cost of bevacizumab (10 mg/kg) plus IFNalpha (9, 6 or 3 million IU [MIU]) versus sunitinib (50 mg) as first-line therapies for advanced or metastatic clear-cell RCC. The average cost per treated patient (year 2010 values) was assessed for the two treatment options at 11 months (median PFS). RESULTS: Assuming a PFS of 11 months for both treatment options, bevacizumab + IFNalpha (9 MIU) would be a lower cost strategy (cost savings of ?2052 per patient) than sunitinib. This difference arises mainly from the reduction in the acquisition cost of bevacizumab to the NHS (risk-sharing agreement). The cost advantages for bevacizumab would increase in parallel with a reduction in IFNalpha dosing; for example, with IFNalpha 6 MIU the corresponding cost savings would be ?4185, and with 3 MIU the cost advantage would be ?6320 per patient. CONCLUSION: This analysis suggests that bevacizumab + IFNalpha is a cost-saving alternative to sunitinib in the treatment of first-line metastatic RCC. Its superior safety profile also meant that the cost of managing adverse events was lower for bevacizumab + IFNalpha than for sunitinib. PMID- 21627340 TI - Bevacizumab-induced hypertension: pathogenesis and management. AB - Bevacizumab, a recombinant humanized monoclonal antibody targeting the vascular endothelial growth factor (VEGF), has been approved in the US as first- and second-line treatment of colorectal cancer and in the first-line treatment of advanced non-small cell lung cancer. The US FDA has also granted approval for the use of bevacizumab for the treatment of patients with metastatic renal cell carcinoma and glioblastoma, and in Europe, it is also approved in metastatic breast cancer in combination with paclitaxel. Bevacizumab is under investigation in the first-line and adjuvant setting of almost all types of solid tumors. However, anti-VEGF therapy is associated with significant toxicity. The incidence of grade 3-4 hypertension differs among the various malignancies in which bevacizumab is administered, possibly because of drug interactions with co administered chemotherapy drugs. Hypertension appears to be dose dependent, and it is under investigation as a biomarker for VEGF inhibition efficacy. There are three main theories concerning the underlying pathophysiology: (i) the nitric oxide theory; (ii) the renal impairment theory; and (iii) the pre-eclampsia-like theory. The correct evaluation of the levels of hypertension is of critical importance and home blood pressure monitoring seems to be the most effective technique. A baseline assessment and follow-up monitoring of blood pressure is considered necessary for all patients receiving bevacizumab. There are no evidence-based recommendations regarding which antihypertensives are more appropriate for the management of bevacizumab-related hypertension. It has been suggested that the benefits from antihypertensive treatment are largely independent of the drugs used, as long as they adequately lower blood pressure. Randomized prospective studies are necessary to provide data that will be useful for the development of specific guidelines for the management of bevacizumab related hypertension. In the meantime, treatment of anti-VEGF-induced hypertension should follow current guidelines for diagnosis and management of hypertension in general. PMID- 21627341 TI - Neovascular age-related macular degeneration: opportunities for development of first-in-class biopharmaceuticals. AB - Age-related macular degeneration (AMD) is a condition that may cause blindness. The prevalence of the disease in the Western world is estimated at 1-2% of the population. Over the past decade, treatment of neovascular AMD has been shifting from destruction of newly formed blood vessels towards inhibitors that silence the vascular endothelial growth factor (VEGF) pathway. Such agents are often first-in-class biopharmaceuticals that benefit from the fact that they can be locally administered in an immune-privileged environment with slow clearance. These new VEGF pathway inhibitors have improved therapeutic effects over conventional treatment and have promoted the identification of novel targets for inhibition of AMD angiogenesis. This review describes the rationale behind the shift from conventional to current treatment options and discusses investigational, most notably biopharmaceutical, drugs that are in clinical trials. It also provides possible points for improvement of these treatments, specifically regarding their delivery. PMID- 21627334 TI - Mammalian triacylglycerol metabolism: synthesis, lipolysis, and signaling. PMID- 21627342 TI - Imatinib: as adjuvant therapy for gastrointestinal stromal tumor?: profile report. PMID- 21627343 TI - Temsirolimus in relapsed and/or refractory mantle cell lymphoma?: profile report. PMID- 21627344 TI - Hot ortho-biologic topics at AAOS 2011?: platelet-rich plasma and related growth factors generate excitement. AB - Several hot topics relating to ortho-biologics were discussed at the 2011 Annual Meeting of the American Academy of Orthopedic Surgery (AAOS) in San Diego this February. Injecting a patient's own platelet-rich plasma (PRP) prior to orthopedic surgery was an important topic, and had its own forum devoted to debating its uses and merit. PRP use has been promoted by equipment companies such as MTF Sports Medicine, Biomet, and Arteriocyte, but others are likely to take advantage of the trend of increasing PRP use by developing a proprietary injectable that mixes PRP with certain growth factors. One possible addition would be a recombinant platelet-derived growth factor (rhPDGF-BB, becaplermin) being developed by BioMimetic Therapeutics for its bone graft product. On the topic of viscosupplementation, the US's only single-injection product, Genzyme's SynviscOne(r), was noticeably missing from the exhibit hall at AAOS, but an abstract comparing the single- and multiple-injection viscosupplementation techniques demonstrated that single-injection acts faster and is longer lasting. New bone morphogenetic protein formulations may improve healing of bone fractures. Molecular diagnostics may be used to predict periprosthetic joint infection, allowing orthopedic medicine to be more personalized. A diagnostic that can be used on a large scale has not yet been identified. ? Adapted and reproduced from Hoggatt J. Hot Ortho-Biologic Topics at AAOS 2011: Platelet-Rich Plasma and Related Growth Factors Generate Excitement. inThought Research, 2011 Feb 28. PMID- 21627345 TI - Belimumab efficacy is 'mild' but market potential still great?: anticipating us approval of the first lupus drug since 1957. AB - Speaking for the inThought Expert Discussion Series in February, Dr George Tsokos mirrored the opinion of many lupus thought leaders: Human Genome Sciences (HGS) and GlaxoSmithKline's (GSK) belimumab (Benlysta(r)) was likely to be approved by the US FDA and, despite modest efficacy, will be used by a large proportion of lupus patients. Dr Tsokos praised HGS and GSK's clinical trial program for belimumab, noting that huge trials and unique trial endpoints were needed to demonstrate the drug's efficacy, allowing it to succeed where so many other lupus drugs have failed. Still, belimumab's trial design may not become standard in future lupus trials - questions about identification of appropriate lupus patients with active disease, trial endpoints, and subgrouping lupus patients remain. Although Dr Tsokos does not expect other agents currently being tested in lupus trials to be significantly more efficacious than belimumab, his research suggests that significantly better results could be obtained using agents targeting interleukin-17, spleen tyrosine kinase (SYK), and calcium/calmodulin dependent protein kinase type IV (CaMKIV). In line with Dr Tsokos' comments and consistent with inThought's outlook for belimumab, the US FDA granted approval for belimumab in March 2011, making it the first new lupus drug to be approved in more than 50 years. inThought projects US sales of $1.1 billion for belimumab by 2017. ? Adapted and reproduced from Weintraub B. Benlysta Efficacy is "Mild" but Market Potential Still Great: Anticipating U.S. Approval of the First Lupus Drug Since 1957. inThought Research, 2011 Feb 24. PMID- 21627347 TI - Novel chemotherapy using histone deacetylase inhibitors in cervical cancer. AB - Since epigenetic alterations are believed to be involved in the repression of tumor suppressor genes and promotion of tumorigenesis in cervical cancers, novel compounds endowed with a histone deacetylase (HDAC) inhibitory activity are an attractive therapeutic approach. In this review, we discuss the biologic and therapeutic effects of HDAC inhibitors (HDACIs) in treating cervical cancer. HDACIs were able to mediate inhibition of cell growth, cell cycle arrest, apoptosis, and the expression of genes related to the malignant phenotype in a variety of cervical cancer cell lines. Furthermore, HDACIs were able to induce the accumulation of acetylated histones in the chromatin of the p21WAF1 gene in human cervical carcinoma cells. In xenograft models, some HDACIs have demonstrated antitumor activity with only few side effects. Some clinical trials demonstrate that HDACI drugs provide an important class of new mechanism-based therapeutics for cervical cancer. In this review, we discuss the biologic and therapeutic effects of HDACIs in treating cervical cancer, especially focusing on preclinical studies and clinical trials. PMID- 21627348 TI - Unsuspected genital tract malignancy discovered during or after gynecologic surgery. AB - Preoperative counseling is a fundamental process in surgical practice. Although uncommon, discordance between preoperative and postoperative diagnoses has been observed in surgical practice. This would be a major concern if a serious condition such as malignant disease is noted incidentally. Encountering unexpected cancers during or after an operation may result in suboptimal treatment performed because of the potential of failure to follow standard treatment guidelines for such cancer. In addition, failing to prepare patients for a possibility of unsuspected cancer is an extremely difficult situation and may complicate the relationship with the surgeon. This article focused on the incidence and major causes of unsuspected genital tract malignancies found during or after gynecologic surgery. PMID- 21627349 TI - Causative relationship between diabetes mellitus and breast cancer in various regions of Saudi Arabia: an overview. AB - The unwarranted connection between diabetes mellitus and breast cancer has gained new ground in recent years. Breast cancer in Saudi females accounts for approximately 21% of all cancers and the prevalence of diabetes mellitus (DM) in Saudi females is also 21.5%. DM is diagnosed in the age group of 30+ years with possible exposure to predisposing factors like hyperinsulinemia and obesity at younger age. Further, 12% of the breast cancer cases are diagnosed in the young females aged 20-34 years. Despite the readily available access to healthcare facilities in the Kingdom, a large number of diabetics, approximately 27.9%, were unaware of having diabetes mellitus. This subpopulation is quite susceptible of developing breast cancer at later age. This review discusses common etiological and predisposing factors for breast cancer and diabetes, regional distribution and possible correlation of diabetes and cancer in Saudi Arabia. PMID- 21627350 TI - 4'-Hydroxycinnamaldehyde from Alpinia galanga (Linn.) induces human leukemic cell apoptosis via mitochondrial and endoplasmic reticulum stress pathways. AB - Rhizomes of Alpinia galanga (Linn.) or 'Kha' in Thai are used in food and as folk medicine in South and Southeast Asia. The aims of this study were to identify the mechanism of cell death of human leukemic HL-60 and U937 cells induced by 4' hydroxycinnamaldehyde (4'-HCA) isolated from A. galanga. 4'-HCA was cytotoxic to both cell lines in a dose-dependent manner (p<0.05) as demonstrated by MTT assay. Apoptosis induced by 4'-HCA was demonstrated by a variety of methods: visualization of propidium iodide (PI)-stained cells under fluorescence microscope, detection of subdiploid cells by PI-staining and flow cytometry, and assay of active caspase-3 using a specific fluorogenic substrate. 4'-HCA-treated cells (10 and 50 MUg/ml for 4 h) showed significant increase in reactive oxygen species production and decreased mitochondrial transmembrane potential as detected by dichlorohydrofluorescein diacetate and 3,3'-dihexyloxacarbocyanine iodide respectively, together with flow cytometry. The apoptotic death involved cytochrome c release, increase in Bax level and concomitant decreases in levels of Bcl-2 and Bcl-xL (using Western blotting), and elevation in cytosolic and mitochondrial Ca2+ contents (using compartment-specific fluorescent Ca2+ dyes). These results indicate that 4'-HCA induces apoptosis of human leukemic cell through a combination of mitochondrial and ER stress pathways. PMID- 21627351 TI - Impact of lymph node metastases in esophageal carcinoma patients is independent of patient age. AB - OBJECTIVE: The purpose of the present study was to define the clinicopathological features and prognosis of esophageal cancer. METHODS: Between 2004 and 2009, 128 patients with esophageal cancer were enrolled in a retrospective database and divided into two groups on the basis of number positive lymph nodes with the cut off as four. RESULTS: The findings for 18 patients (14.0%) Group A were compared with those of 110 patients Group B. In the group A, there were significantly more women (12/6 vs. 54/56, P < 0.001). In both groups, the most frequent histological morphology was squamous cell carcinoma (83% and 75%, respectively), although the percentages were significantly different (P < 0.005). In the group A, lesions were more frequently located in the middle one-third of the esophagus than in the group B (61% vs. 28%, P < 0.001). Group A was more likely to be Stage IIa. Survival rates in group A patients at 5 years after resection were 15.8%, similar to those in group B patients (12.1%, difference not significant). Local lymph node metastases and microscopic residual tumor at the line of resection were also more prevalent in the young patients, but not to a statistically significant degree. CONCLUSIONS: These findings suggested that the clinical and pathologic features of carcinomas of the esophagus in young patients do not significantly differ from those in older patients. PMID- 21627352 TI - Roles of diet, lifetime physical activity and oxidative DNA damage in the occurrence of prostate cancer among men in Klang Valley, Malaysia. AB - BACKGROUND: There is a paucity of information on risk factors of prostate cancer, especially those related to dietary and lifestyle among Asian populations. OBJECTIVE: This study aimed to determine the relationship between dietary intake (macronutrients, fruits, vegetables and lycopene), lifetime physical activity and oxidative DNA damage with prostate cancer. DESIGN: A case control study was carried out among 105 subjects (case n=35, control n=70), matched for age and ethnicity. Data on sociodemographic, medical, dietary intake, consumption of lycopene rich food and lifetime physical activity were obtained through an interview based questionnaire. Anthropometric measurements including weight, height and waist hip circumferences were also carried out on subjects. A total of 3 mL fasting venous blood was drawn to assess lymphocyte oxidative DNA damage using the alkaline comet assay. RESULTS: Cases had a significantly higher intake of fat (27.7 +/- 5.5%) as compared to controls (25.1 +/- 5.9%) (p < 0.05). Mean intakes of fruits and vegetables (3.11 +/- 1.01 servings/d)(p < 0.05), fruits (1.23 +/- 0.59 servings/d) (p<0.05) and vegetables (1.97 +/- 0.94 servings/d) were higher in controls than cases (2.53 +/- 1.01, 0.91 ? 0.69, 1.62 +/- 0.82 servings/d). A total of 71% of cases did not met the recommendation of a minimum of three servings of fruits and vegetables daily, as compared to 34% of controls (p < 0.05) (adjusted OR 6.52 (95% CI 2.3-17.8)) (p < 0.05). Estimated lycopene intake among cases (2,339 ? 1,312 mcg/d) were lower than controls (3881 ? 3120 mcg/d) (p< 0.01). Estimated lycopene intake of less than 2,498 mcg/day (50th percentile) increased risk of prostate cancer by double [Adjusted OR 2.5 (95%CI 0.99-6.31)]. Intake of tomatoes, watermelon, guava, pomelo, papaya, mango, oranges, dragon fruit, carrot, tomato sauce and barbeque sauce were higher in controls compared to cases. Intake of tomato sauce of more than 2.24 g/d (25th percentile), papaya more than 22.7 g/d (50th percentile) and oranges more than 19.1g/h (50th percentile) reduced prostate cancer risk by 7.4 (Adjusted OR 7.4 (95% CI 1.17-46.8)), 2.7 (adjusted OR 2.75 (95% CI 1.03-7.39)) and 2.6 times (adjusted OR = 2.6 (95% CI=1.01-6.67)), respectively (p < 0.05 for all parameters). No oxidative damage was observed among subjects. Past history of not engaging with any physical activities at the age of 45 to 54 years old increased risk of prostate cancer by approximately three folds (Adjusted OR 2.9(95% CI = 0.8-10.8)) (p < 0.05). In conclusion, low fat diet, high intake of fruits, vegetables and lycopene rich foods and being physical active at middle age were found to be protective. Thus, it is essential for Malaysian men to consume adequate fruits and vegetables, reduce fat intake and engage in physical activity in order to reduce prostate cancer risk. PMID- 21627353 TI - Antitumor and cytotoxic activities of methanol extract of Indigofera linnaei Ali. AB - Methanol extract of Indigofera linnaei (MEIL) was investigated for antitumor, cytotoxic and antioxidant activities against transplantable tumors and human cancer cell lines. In vitro cytotoxicity was evaluated in HeLa, Hep-2, HepG-2, MCF-7, HT-29, Vero and NIH 3T3 cells by MTT assay and in vivo antitumor activity with Ehrlich ascites carcinoma (EAC) and Dalton's ascites lymphoma (DLA) tumor bearing mice. Activity was measured by monitoring the mean survival time, effect on hematological parameters, antioxidant enzyme levels and solid tumor volume. The extract exhibited strong in vitro cytotoxicity against all the tested cancer cell lines, but it was found to be safe with normal cells. MEIL at the dose of 200 and 400 mg/kg, significantly increase the mean survival time (P<0.001), exerted a protective effect on the hemopoietic system, demonstrated in vivo antioxidant activity and significantly reduce solid tumor volume (P<0.01). These results show a significant antitumor and cytotoxic effect of MEIL against EAC, DLA and human cancer cell lines and support the ethnomedical use of Indigofera linnaei. PMID- 21627354 TI - Prevalence and of smoking and associated factors among Malaysian University students. AB - OBJECTIVE: The objectives were to determine the prevalence and associated factors for smoking among university students in Malaysia. METHODOLOGY: A cross-sectional study was conducted among 199 students in the period from December of academic year 2009 until April of academic year 2010 in Management and Science University (MSU), Shah Alam, Selangor, Malaysia. The questionnaire was distributed randomly to all faculties of MSU by choosing one of every 3 lecture rooms, as well as the library and cafeterias of the campus randomly by choosing one from every 3 tables. Questions concerned socio-demographic variables, knowledge, attitudes and practice toward smoking. Participant's consent was obtained and ethical approval was provided by the ethics committee of the University. Data entry and analysis were performed using descriptive statistics, chi square test, Student t- test and logistic multiple regression with the SPSS version 13.0, statistical significance being concluded at p < 0.05. RESULTS: About one third of students were smokers (29%). The most important reason of smoking was stress (20%) followed by 'influenced by friends' (16 %). Prevalence of smoking was significantly higher among male and those in advanced semesters (p = >0.001, p = 0.047). Smokers had low level of knowledge (p < 0.05), had wrong beliefs on smoking (p < 0.05), and negative attitude toward tobacco control policies compared to non smokers (p < 0.05). On multiple logistic regression, significant predictors of smoking in the model were gender (p = 0.025), age (p = 0.037), semester of study (p = 0.025) and attitude toward smoking (p < 0.001). CONCLUSION: This study found that 29% of university students were smokers. Males and students in advanced semesters were more likely to smoke. The results provide baseline data to develop an anti smoking program to limit smoking in the university by implementing policies against smoking. PMID- 21627355 TI - Immunohistochemical profile of breast cancer patients at a tertiary care hospital in South India. AB - AIMS: 1) To evaluate the estrogen receptor(ER), progesterone receptor (PR) and Her-2 /neu expression in invasive breast carcinomas by immunohistochemistry and 2) to compare the pattern of expression with clinico-pathological parameters like patient's age, tumor size, mitotic index, histological type and grade and lymph node metastasis. METHODS: This is a retrospective study of 321 female invasive breast carcinomas diagnosed in the Department of Histopathology, Apollo Speciality Hospital, Chennai from January 2009 to June 2010. RESULTS: The age of the patients ranged from 24 to 99 years, with a mean of 53.8, and the majority of the tumors were T2 (83.8% in range of 2-5 cms), predominantly histological grade 2 (57.3%), followed by grade 3 (33.3%). ER, PR and Her-2/neu expression was seen in 59, 51 and 27% of cases respectively. Triple-negative breast cancers constituted 25 % of our cases. We also found characteristic associations between hormonal receptor and Her-2/neu expression and various clinico-pathological parameters. CONCLUSIONS: The hormonal receptor expression appears to be lower in the Indian population compared to the West. A significant proportion of tumors in our study with Her2/neu overexpression also showed ER and PR positivity. Triple negative breast tumors were most commonly grade 3, in women aged more than 50 years. PMID- 21627356 TI - Risk of the contralateral mucosa in patients with oral potentially malignant disorders. AB - BACKGROUND: It is known that abnormal changes may occur in any part of the oral mucous membrane exposed to a carcinogen. Therefore patients with oral potentially malignant disorders (PMDs) are at risk of developing similar lesions at multiple sites. OBJECTIVES: To determine the risk of the contralateral mucosa in patients presenting with oral PMDs. MATERIALS AND METHODS: Sixty individuals with PMDs were selected for this study. These comprised 32 (53.3%) Indians, 23 (38.3%) Chinese, four (6.7%) Malays and one (1.7%) Nepalese. All selected cases had histopathological confirmation of their primary existing lesion as inclusion criteria. Cases that subsequently presented with a lesion in the corresponding anatomical site also underwent scalpel incisional biopsy on this second lesion to verify its diagnosis. The remaining cases that presented with unilateral PMDs at the time of study were subjected to a cytobrush biopsy on the normal looking contralateral mucosa. RESULTS: A total of 70 primary PMDs were detected in 60 patients. The most common PMD found was oral lichen planus (n=40, 57.1%). Of the 60 patients studied, 28 (46.6%) exhibited bilateral lesions either synchronously (n=21, 35.0%) or metachronously (n=7, 11.6%). The remaining cases that had undergone cytobrush biopsy on the corresponding anatomical site yielded normal cytological results. CONCLUSIONS: Present findings demonstrated that patients presenting with PMDs in the upper aerodigestive tract are at a greater risk of developing a second lesion most probably in the contralateral anatomical site. PMID- 21627357 TI - Measuring quality of life in thai women with breast cancer. AB - Breast cancer has become a commonly diagnosed disease among Thai women in the last decade, despite the fact that Thai women generally have a lower rates than their Western counterparts. With the rising incidence and survival rates, it is crucial for nurses to look at the long term quality of life of these patients. A broad range of instruments have been used in clinical trials among breast cancer patients in oncology, like the EORTC questionnaire including the general quality of life questions (QLQ-C30) and the breast cancer module (QLQ-BR23), and the FACT B questionnaire consisting of both a generic part (FACT-G) and a breast cancer specific module. They have been shown to have good validity and reliability properties both for the English original and translations into various languages including Thai. A few studies on quality of life in Thai context exist, covering quality of life in women with breast cancer. Therefore, the purpose of this study was to find which standard measure of common Western quality of life scales is appropriate to assess quality of life in Thai women with breast cancer. Results revealed the Thai version of EORTC QLQ-C30 and FACT-G questionnaires to be reliable and valid to assess quality of life in general. The best fit for measuring quality of life in Thai women with breast cancer during adjuvant treatment should be the EORTC QLQ-C30/-BR23. PMID- 21627358 TI - Prevalence and distribution of high risk human papillomavirus (HPV) Types 16 and 18 in Carcinoma of cervix, saliva of patients with oral squamous cell carcinoma and in the general population in Karnataka, India. AB - BACKGROUND: In view of conducting HPV vaccination in India it is most important to understand the prevalence of HPV genotypes in this population, not only in squamous cell carcinoma of cervix and oral cavity but also in the general population. In this study we explored the prevalence and distribution of high risk HPV types 16 and 18 in carcinoma of cervix, saliva of patients with oral squamous cell carcinoma and in general population in Karnataka. METHODS: Cervical cancer specimens after punch biopsy (n=60) were obtained from women attending Karnataka Institute of Medical Sciences and Karnataka Cancer Therapy and Research Institute, Hubli (KCTRI). Saliva rinse of (n=34) OSCC patients from KCTRI and (n=396) normal individuals from different regions of North Karnataka, were collected and PCR based high-risk HPV genotyping was carried out. RESULTS: Using consensus PCR primers it was observed that 96.7% patients were infected with HPV irrespective of specific type in cervical cancer. Among them, HPV 16 was observed in 89.7%, HPV 18 in 86.2% and both HPV 16 and 18 in 79.3% patients. In OSCC, 70.6% were positive for HPV, among which HPV 16 prevalence was observed in 45.8%, HPV 18 in 54.2%, and HPV 16 and 18 multiple infection in 4.18%. In general population, HPV prevalence was observed in 84.4%. Among them, HPV 16 was observed in 2.75% and HPV 18 in 22.0% patients. In general population, multiple infection with HPV 16 and 18 was not observed but 75.3% were found to be infected by HPV genotypes other than HPV 16 and 18. CONCLUSIONS: Our study reveals that multiple infection of HPV 16 and 18 is quite high in cervical cancer and in case of OSCC, it was in conformity with the other studies. In general population HPV 18 prevalence was observed to be high. With this, we can conclude that both HPV 16 and 18 vaccinations will reduce the burden of cervical cancer and OSCC in Karnataka. PMID- 21627359 TI - High p16 expression predicts a positive response to chemoradiotherapy in stage IVa/b head and neck squamous cell carcinoma. AB - BACKGROUND: The objective of this study was to evaluate the effect of p16 expression on response to chemoradiation in stage IVa/b head and neck squamous cell carcinoma (HNSCC) patients. METHODS: We retrospectively identified 64 patients with stage IVa/b HNSCC who received chemoradiation. Eligibility criteria included presence of biopsy-proven stage IVa/b HNSCC without a prior history of chemotherapy or radiotherapy. Immunohistochemistry was used to assess p16 protein expression in pretreatment biopsy specimens. RESULTS: Of the 64 patients, 38 showed high p16 expression, and 50 patients responded to chemoradiotherapy, 32 exhibiting a complete and 18 a partial response. Response was significantly associated with p16 expression (P < 0.001) and multivariate analysis indicated that that p16 expression (HR: 2.62, 95%C.I.: 1.14-6.06; P=0.024) was an independent prognostic factor for overall survival. CONCLUSIONS: High p16 expression predicts a better response to chemoradiation in patients with stage IVa/b HNSCC. PMID- 21627360 TI - High expression of CXCR7 combined with Alpha fetoprotein in hepatocellular carcinoma correlates with extra-hepatic metastasis to lung after hepatectomy. AB - The lung is the most frequent metastatic site of hepatocellular carcinoma (HCC), negatively impacting on survival rates. In this study, we evaluated the prognostic role of the chemokine receptor CXCR7 in lung metastasis of HCC after hepatectomy, using immunohistochemical detection on tissue microarrays of HCCs, with and without lung metastasis. Using three categories based on staining characteristics, patients with high CXCR7 expression demonstrated a shorter time to development of lung metastasis compared with patients with low CXCR7 expression (log-rank test) with no effet on overall survival. Analysis of tissue adjacent to tumor showed patients with microvascular invasion to have higher CXCR7. Stratification based on alpha fetoprotein level >20 ng/ml also showed high expression of CXCR7 to be a strong independent prognostic factor. These findings suggest that high expression of CXCR7 in HCCs with elevated alpha fetoprotein levels correlates with metastasis to lung and poor survival after hepatectomy, indicating potential use as a prognostic factor. PMID- 21627361 TI - Suppression of DMBA/croton oil-induced mouse skin tumor promotion by Ardisia Crispa root hexane extract. AB - Ardisia crispa (Family: Myrsinaceae) has been used as a traditional medicine for various ailments. Previous studies showed that Ardisia crispa possesses antimetastatic and anti-inflammatory properties. Nevertheless, research done on the plant is still limited. Therefore, the present study was designed to evaluate the suppression effect of Ardisia crispa root hexane (ACRH) extract on 7, 12 dimethylbenz (alpha) anthracene (DMBA)-induced mice skin tumor promotion in ICR mice with topical application twice weekly for 10 weeks. Results showed significant difference between treatment groups (mice treated with 30 mg/kg, 100 mg/kg and 300 mg/kg of ACRH extract; denoted as group I, II and III respectively) for tumor incidence and tumor burden (P<0.05). Significant reduction in tumor incidence (20%), tumor burden (1.5 +/- 0.50), tumor volume (2.49 +/- 1.70) and delayed latency period of tumor formation was observed in group I (30 mg/kg) in comparison to carcinogen control. This study indicates that ACRH extract could be a promising skin tumor promotion suppressing agent at a lower dosage (30 mg/kg). Further studies are required to elucidate the underlying mechanism(s) leading to this effect. PMID- 21627362 TI - Prevalence and risk of leukemia in the multi-ethnic population of North Karnataka. AB - BACKGROUND: The aim of this study was to analyze the trends in the incidence rates of four major types of leukemia in the population of North Karnataka, which accounts for the 2.5% of the whole population of India. Due to the lack of any nationwide leukemia screening program, the majority of the people are not aware of the disease. Epidemiological study can play a vital role in understanding the occurrence and outcome of the disease. PATIENTS AND METHODS: Focusing on variables like age, sex, race, blood group and lifestyle habits, detailed reports of 417 males and 230 females (M:F ratio 1.8:1) were collected from different hospitals of North Karnataka and analyzed for their risk of leukemia. RESULTS: Compared to female patients, Hindu males were found to have greater risk of occurrence of leukemia (p=0.0333). The males of scheduled caste (SC) and Lingayat communities showed a high risk than other communities (p=0.000). The occurrence of AML showed a significant relationship with ABO blood groups (p=0.0090). The frequency of leukemia is quite high in Belgaum district when compared to others districts of North Karnataka and totally absent in Bidar district. The reasons need precise molecular and genetical studies of the population. CONCLUSIONS: The localized communities of Lingayat and SCs needs to be further studied to get a better understanding of the higher risk of occurrence of leukemia in males. Moreover, since the spectrum of cancer epidemiology seen in India is different from that in developed countries more emphasis should be placed on better development of regional and national registries. PMID- 21627363 TI - Chemopreventive effect of hydroethanolic extract of Euphorbia neriifolia leaves against DENA-induced renal carcinogenesis in mice. AB - The present study was conducted to investigate the chemopreventive effects of hydro-ethanolic extract of Euphorbia neriifolia (EN) on N-nitrosodiethylamine (DENA) induced renal cancer in male Swiss albino mice. Animals were pretreated with EN extract (150 and 400 mg/kg body weight; p.o) and butylated hydroxyanisole (BHA) as a standard (0.5% and 1% BHA p.o) both for two week prior to the administration of single dose of DENA (50 mg/kg body weight; p.o). Various in vivo antioxidant biochemical parameters like lipid peroxidation (LPO), superoxide dismutase (SOD), and catalase (CAT) were evaluated to determine the reno protective and antioxidant activity of EN. DENA increased oxidative stress through increase in LPO and decrease in antioxidant enzymes (SOD, and CAT). The EN extract significantly restored the antioxidant enzyme level in the kidney and exhibited significant dose dependant protective effect against DENA induced nephrotoxicity, which can be mainly attributed to the antioxidant property of the extract. This study rationalized the ethno-medicinal use of EN for protection against renal cancer. PMID- 21627364 TI - Anaplastic large cell lymphoma: the most common T-cell lymphoma in pakistan. AB - OBJECTIVE: To study the prevalence and immunohistochemical profile of the subtypes of anaplastic large cell lymphomas (ALCLs) at a major referral center of Pakistan. METHODS: Epidemiological data for all mature T-cell non-Hodgkin's lymphoma (NHL) diagnosed between 1st January 2005 and 30th June 2010 at the Aga Khan University Department of Histopathology were reviewed and analyzed with SPSS v17.0. RESULTS: A total of 178 specimens were diagnosed as mature T- and NK- cell NHL during the period. Of these 100 (56.2%) were diagnosed as systemic ALCL. These tumors were of either T- or null-cell type with consistent (100%) expression of CD30 (Ki-1). Forty three (43%) cases were further classified as ALK positive, fifty (50%) as ALK negative and seven (7%) were not tested for ALK expression. The mean age of the ALK positive group was 26.7 years as compared to the ALK negative of 35.6 years. The gender ratio of ALK positive cases was 2.3:1 (M:F) as compared to the 2.5:1 ratio seen in the ALK negative cases. There were no significant differences in the nodal and extra-nodal involvement patterns between ALK+ and ALK- groups but epithelial membrane antigen was positive more often in the ALK positive group. CONCLUSION: Compared to other published studies, this proportion of ALCL within the mature T- and NK- cell lymphoma category was found to be alarmingly high and calls for attention. Further studies should be conducted in our region, which in turn would enable clinicians to successfully battle against this neoplastic disease. PMID- 21627365 TI - Nutrition and cancer prevention: knowledge, attitudes and practices among young Malaysians. AB - OBJECTIVE: The objective of this study was to determine the knowledge, attitudes and practices of university students toward nutrition related to cancer prevention. METHODOLOGY: A total of 396 students from the Management and Science University (MSU) participated during the semester of March 2010. Stratified random sampling was used and consent was obtained before the questionnaire was distributed. ANOVA and the t-test were used for the univariate analysis and multiple linear regression was used for the multivariate analysis. RESULTS: The participants ages ranged from 18 to 27 years (Mean +/- SD = 23.3 +/- 1.57), more than half being female (62.4%). The majority were 23 years old or younger, single, Malay and from non-Medical and Health Science faculties and with a family monthly income of less than 10,000 Ringgits Malaysia(79.5%; 99%, 65.9, 52.5%, 63.9%; respectively). Only 18.4% of participants reported a family history of cancer. About 32.1% had a medical check-up in the previous 12 months and 17.4% were smokers. Multivariate analysis showed the faculty type to be significantly associated with knowledge of cancer prevention (p = 0.04). Regular medical check ups were associated with attitudes and practices of cancer prevention (p = 0.04, p=0.003 respectively), the latter being significantly influenced by sex, family history of cancer and smoking (p = 0.034, p=0.013, p=0.002; respectively). CONCLUSION: The majority of participants had poor knowledge of nutrition as related to cancer prevention. Attention should be given to regular medical check ups, awareness of family history and smoking influence. PMID- 21627366 TI - Testicular cancer awareness and testicular self-examination among university students. AB - BACKGROUND: This study was planned to determine the knowledge of testicular cancer (TC), risk factors and testicular self-examination (TSE) among university students. METHODS: A survey study was conducted at a university located in Ankara, Turkey with 634 male students. RESULTS: Almost half of them (44%) heard TC during their education and life but majority of participants has lack of knowledge about sign and symptoms of TC. Only 5.9 % of them (n = 38) indicated they received information on TSE and 17.7 % have performed the practice of TSE before; only one in a forth (n=21) performed monthly. The reason for not doing TSE was mostly (83.4%) 'not having knowledge' and 'not seen as important' (55.7%). CONCLUSIONS: Adding the education on TC, risk factors and TSE into their curriculum is proposed. PMID- 21627367 TI - Pretreatment levels of serum vascular endothelial growth factor do not correlate with outcome in patients with locally advanced cervical cancer. AB - OBJECTIVE: To evaluate pretreatment levels of serum VEGF in locally advanced cervical cancer patients, and assess any association with clinocopathological parameters and response to radiotherapy. METHODS: Patients with histologically proven and diagnosed locally advanced cervical cancer or stages IIB-IVA were included in this study. Blood serum was obtained by peripheral venous puncture about 24 hours before the beginning of radiotherapy. All patients were followed up at one and three month intervals from the last day of the complete treatment for evaluating the responses to radiotherapy. RESULTS: Mean age of the 40 patients was 52.8 +/- 11.1 years. Sixty percent were in stage IIB and 90% had squamous cell carcinoma. The median pretreatment level of serum VEGF was 611.3 pg/ml (0.00-4,067 pg/ml). The pretreatment levels of serum VEGF did not correlate with stage (p=0.75), tumor histology (p=0.91), tumor size (p=0.46) or tumor characteristics (p=0.49). Almost all patients received concurrent chemoradiation as a curative treatment, with a complete response found in 94.9%. Values for patients who were completed response was rather lower than in patients with persistent disease, but without statistical significance (581.4 pg/ml vs 759.6 pg/ml, p=0.37). CONCLUSION: Pretreatment levels of serum VEGF do not correlate with clinicopathological factors or response to radiation therapy. PMID- 21627368 TI - Incidence and current trends of colorectal malignancies in an unscreened, low risk Pakistan population. AB - INTRODUCTION: Karachi falls into a low risk region for colorectal cancer (CRC). The incidence rate is known but detailed epidemiology and pathology data regarding the disease are not available. The aim of this study was to describe CRC with reference to incidence, gender, topographic sub-site, tumor morphology, grade and stage at diagnosis and to determine the trends of incidence. MATERIALS AND METHODS: Four hundred and seventy three cases of colorectal cancer submitted to the Karachi Cancer Registry for Karachi South, years 1995-2002 were reviewed. Cases were analyzed in two time periods (1995-7 and 1998-2002) to facilitate the study of time trends. RESULTS: A total of 151 CRC cases were registered during period one [86 (57%) males; 65 (43%) females] and 322 cases [210 (65%) males; 112 (35%) females] in period two. Age standardized rate (ASR) world per 100,000, crude incidence rate (CIR) and relative frequency in period one were 5.3, 3.2 and 4.1% in males and 5.5, 3.2 and 3.2% in females respectively. Corresponding figures for period two were 7.1, 4.5 and 4.7% for males and 5.2, 2.8 and 2.7% in females. The male, female ratio was equal for colon (1:1). Men had more rectal cancers (2:1) and overall CRC (1.7:1). The mean age of the patients varied with sub-site and gender from 43.7 years to 51.2 years. Cancers of the rectum presented at a relatively earlier age. Less then 5% of the cases were diagnosed in adolescents, 50% above 50 years of age and only 30% above 60 years. The ratio under-40 to above-40 for CRC patients was 0.3, which is much higher than the international average, indicating a younger age group at risk. The first cases were observed in adolescents (15-19 years) and a peak was observed in the seventh decade. Colon to rectum ratio was 1:1 in males and 2:1 in females. Most cases presented with advanced disease, though some down staging was observed in period 2 (1998-2002). CONCLUSION: The current low but increasing incidence (especially in men), the younger age and advanced stage of CRC at diagnosis reflects a low risk, unscreened population. With existing prevalence of high risk factors in Pakistan, the low CRC incidence may be an artifact. There are concerns that an aging population over the next decade and changing lifestyle patterns may translate into a higher CRC incidence. Screening must be considered as part of the health sector planning for the future and include the high risk younger age groups. PMID- 21627369 TI - Younger women are affected by breast cancer in South India - a hospital-based descriptive study. AB - INTRODUCTION: Breast cancer is one of the most common cancers in the world among women and its effective control depends upon sufficient knowledge about its epidemiology at the regional as well as global levels. MATERIALS AND METHODS: A record based descriptive study was conducted in a tertiary care hospital situated in coastal Karnataka in South India. Cases included were all histopathologically diagnosed cases of breast cancer not otherwise specified among females that were treated in the hospital in the 3 years period of 2005-2007. A total of 112 cases were found and their data retrieved and analyzed using SPSS. RESULTS: Mean age of the subjects was 45 years (SD=10.4) (range 23 -70). Age distribution showed two peaks at 35-39 years and 50-54 years. Some 12 (11%) patients had a previous history of breast cancer and 9 (8%) had a family history. Among the patients who had history of breast cancer in the past, 61.5% were during the pre-menopausal age. CONCLUSION: The age affected by breast cancer was lower than usually seen in other parts of the world and there was more of premenopausal occurrence which has implications on their own and their relatives' chances of acquiring breast cancer later. The findings show that breast cancer control efforts and recommendations in different parts of the world have to be tailored to the regional epidemiological patterns. PMID- 21627370 TI - Cytogenetic biomonitoring of road paving workers occupationally exposed to polycyclic aromatic hydrocarbons. AB - Road pavement workers are exposed to many known carcinogens in their complex occupational environment. The study makes an attempt to investigate exposure to polycyclic aromatic hydrocarbons (PAH) from the bitumen fumes among the road pavement workers engaged in different pavement sites at Coimbatore, Tamil Nadu and to thereby determine the genotoxic effects associated with it. The study included 36 road pavers and 37 control subjects with similar mean ages, smoking prevalence and alcohol consumption and was analyzed for DNA damage in blood leucocytes by Micronucleus assay (MN) and the Comet assay. The mean urinary 1-OHP concentration in road pavers (1.68 ? 0.93) was significantly higher than in controls (0.55 ? 0.42). The results of MN test and comet assay showed that the mean micronuclei rate in workers was significantly higher than those in controls (P <0.05). The results of our study indicated that the genetic damage was detectable in road paving workers occupationally exposed to bitumen and also demonstrate the high sensitivity of comet assay to assess early oxidative effects induced by exposure to bitumen fumes at low doses and confirm the suitability of urinary 1-OHP as a biomarker of PAH exposure. PMID- 21627371 TI - Impact of triple negative phenotype on prognosis and early onset of breast cancer in Iranian females. AB - BACKGROUND: Breast cancer in Iranian women occurs about a decade earlier than in Western countries. This study sought to evaluate the impact of triple negative phenotype on early onset of ductal cell breast cancer and its prognosis in Iranian females. METHODS: Estrogen and progesterone receptors, Her-2 overexpression and nuclear accumulation of P53 were assessed in sixty surgically resected formalin-fixed paraffin embedded breast invasive ductal carcinomas. They were divided into triple negative and non triple negative phenotypes and impact of the phenotypes were evaluated on prognostic factors of all patients and based on menopausal status. RESULTS: The result showed that the mean age of patients with triple negative breast tumors, especially in postmenopausal group, was significantly lower than with non triple negative phenotypes. Although the latter was significantly associated with higher histological grade, it also showed a significant correlation with smaller size of tumor and a lower rate of axillary lymph node metastasis in young patients. CONCLUSION: The higher rate of breast cancer with triple negative phenotype in Iranian females is a feasible reason for the reported lower mean age of breast cancers. In premenopausal patients, triple negative phenotype reveals a positive impact on prognostic factors, but it is associated with a poorer prognosis in postmenopausal patients. Hence, a distinct ethnic profile of triple negative phenotype in Iranian females is suggested. PMID- 21627372 TI - Overview of the National Cancer screening programme and the cancer screening status in Korea. AB - Organised cancer screening in Korea began in 1999. Operating system has been stabilised, target population have expanded and participation rate has been increased throughout its ten years. Here we present an overview of the organised cancer screening system in Korea and introduce the National Cancer Screening Programme including results from 2002 to 2008. Furthermore, we present the results of the Korea National Cancer Screening Survey, a survey that is representative of the population, from 2004 to 2009. Finally, we discuss our achievements and the future challenges. PMID- 21627373 TI - Associations of polymorphisms in HPC2/ELAC2 and SRD5A2 genes with benign prostate hyperplasia in Turkish men. AB - Benign prostate hyperplasia (BPH) is the most common benign tumor in elderly men for which the HPC2/ELAC2 and SRD5A2 genes are known genetic factors. The HPC2/ELAC2 gene features Ser217Leu and Ala541Thr polymorphisms and the SRD5A2 gene Ala49Thr and Val89Leu polymorphisms. The aim of this study was to examine relationships between these polymorphisms and BPH in Turkish men using amplification by the polymerase chain reaction (PCR) method. Polymorphisms were determined by using restriction fragment length polymorphism (RFLP) with suitable restriction: TaqI?, Fnu4HI, Mwo I and Rsa I. We found statistically significant relationship between the SRD5A2 gene Ala49Thr (OR=2.3; CI 95%, 1.04-5.1; p=0.01<0.05) , but not the other polymorphisms, and BPH. For the first time, our data demonstrate that the correlation between SRD5A2 gene Ala49Thr and polymorphisms is statistically significant in Turkish men with BPH. PMID- 21627374 TI - Nasopharyngeal carcinoma - an update of treatment and acute radiation induced reactions from a tertiary-care hospital in Pakistan. AB - INTRODUCTION: Nasopharyngeal carcinoma (NPC) is a diverse entity with a multi factorial etiology and a distinct racial and geographical distribution. It is curable if diagnosed and treated early. METHODS: This descriptive study covered 30 patients who underwent radiotherapy (RT) for nasopharyngeal malignancies during February 2006 till November 2010 at the Department of Radiation Oncology, Aga Khan University Hospital (AKUH), Karachi. RESULTS: Thirty cases of nasopharyngeal tumors were registered; a case of embryonal rhabdomyosarcoma and another of diffuse large B-cell lymphoma were not included in the final analyses. There were 20 (71.0%) males and 8 (29.0%) females, 2 and 5 being observed in adolescents. The mean age of the male and female patients was 43.7 [SD 20.1] and 30.3 years [SD 17.9], respectively. Nodal involvement was seen in 23 cases and cranial in 8. Almost two-thirds the patients presented with a stage IV disease and all but two received chemotherapy. Electrons or photons were used for 23 cases; low anterior neck field was used in 25 cases. The doses of radiotherapy delivered ranged between 2000cGy to 7400cGy, the lower ones being given for palliation only. Once spinal cord tolerance was reached, electron boost fields were used in order to treat the posterior neck. A nasogastric tube was required for feeding in 2 (10.0%) male and 2 (25.0%) female patients; gastrostomy was needed for 1 (5.0%) patient. Radiotherapy (RT) induced reactions were observed in 90% of the patients, but were mild in most cases. CONCLUSIONS: In the presence of clinician based expertise and technical constraints in Pakistan, nasopharyngeal tumors are still being treated optimally with the need of creating a better awareness in public for early detection. PMID- 21627375 TI - Lack of any relationship between chemotherapy toxicity in non-small cell lung cancer cases and polymorphisms in XRCC1 codon 399 or XPD codon 751. AB - PURPOSE: To examine the association between genetic polymorphisms (at XRCC1codon 399 or XPD codon 751) and chemotherapy related toxicities of non-small cell lung cancer. METHODS: One hundred and fifteen patients with histologically or cytologically confirmed stage IIIB and IV NSCLC recruited from Department of Chemotherapy of Jiangsu Cancer Hospital and Research Institute from 2005 to 2008, to evaluated the occurrence of chemotherapy related toxicities and the association with single nucleotide polymorphisms in XRCC1codon 399 or XPD codon 751. RESULTS: No significant association was observed between grade 0 or grade 1 4 overall toxicity and XRCC1 codon 399 (odds ratio=1.40, 95% confidence interval,0.73-2.66; adjusted odds ratio =1.43, 95% confidence interval,0.71 2.88), or XPD codon 751 genetic polymorphisms (odds ratio =0.87, 95% confidence interval,0.33-2.26; adjusted odds ratio=0.74, 95% confidence interval,0.26-2.13); similar results were found between hematologic, hepatic, gastrointestinal toxicities and XRCC1 399 or XPD 751 genetic polymorphisms. CONCLUSION: No statistically significant association was found between either XRCC1codon 399 or XPD codon 751 genetic polymorphisms and chemotherapy related toxicities. PMID- 21627376 TI - Weight gain and family history of prostate or breast cancers as risk factors for prostate cancer: results of a case-control study in Japan. AB - The increase in the incidence rate of prostate cancer may be associated with changes in lifestyle in Japanese men. Accordingly, we conducted a case-control study to assess risk factors. A total of 117 (82.3%) of the 142 prostate cancer patients asked filled out the self-administrated questionnaires which included items about their lifestyle habits over the period of one or two years before their diagnosis. Four controls per case, namely 468, were randomly selected from resident registries with age and address matched with each case, and 318 controls (69.5%) filled out the same questionnaire as the cases. Data for 277 controls were used for the analysis, excluding 41 subjects with a history of previous cancer. The conditional logistic regression model was utilized for analyzing the individually age and address-matched data, and odds ratios (ORs) and their 95% confidence intervals (95%CIs) were calculated for potential risk factors. Higher body mass index at 20 years of age was marginally significantly associated with a decreased risk (P for trend=0.051), and larger weight gain in adult age was significantly associated with an increased risk (P for trend=0.041). History of prostate cancer in fathers or brothers was significantly associated with an increased risk (OR=9.71, 95%CI 3.59, 26.27), and history of breast cancer in mothers or sisters was also significantly associated with an increased risk (OR=2.70, 95%CI 1.12, 6.49). The recent increase in the incidence rate of prostate cancer may possibly be brought about by an increased proportion of Japanese men with large weight gain in adult age. PMID- 21627377 TI - Human papillomavirus testing for suspected cervical cancer patients from Southern Assam by fast-PCR. AB - World-wide epidemiological studies have shown that cancer of the uterine cervix is the second most common malignant disease in women. Virtually every cervical cancer (99.7%) is HPV-positive, indicating that the presence of HPV is an obligatory element in their development. The present study was conducted by Fast PCR (within 15 min.) based diagnosis of HPV 16 and HPV 18 infection amongst patients of suspected cervical cancer, confirmed by cytological methods. Twelve women, out of a total of fifty studied cases who had positive cervical pap smears (24%) were found to be positive for HPV 16/HPV 18 infection when PCR based technique was applied. The results indicate, perhaps, a greater specificity of PCR based diagnosis, or presence of other HPV subtypes as etiological factors in the present study group confined to Southern Assam. PMID- 21627378 TI - Dietary patterns and risk of colorectal cancer: a factor analysis in uruguay. AB - In the time period 1996-2004, a case-control study of colorectal cancer was conducted in Montevideo, Uruguay. The study included 610 cases and 1,220 controls, frequency matched for age, sex, and residence. All cases were newly diagnosed and microscopically confirmed and controls were drawn from the same hospitals. Controls were submitted to factor analysis (principal components method) and 4 dietary patterns for men (prudent, traditional, Western, drinker) and 3 for women (prudent, Western, drinker) were retained. These were rotated and normalized by the Kaiser method. Scores were applied to all participants (cases and controls) and odds ratios were estimated by logistic regression and polynomial regression. The Western pattern showed an OR of 2.62 (95 % CI 1.36 5.08) for colon cancer among men, and women displayed a similar increase in risk. However, rectal cancer was not associated with this diet, rather being inversely associated with the prudent and traditional patterns among men (OR 0.49, 95 % CI 0.28-0.57 for the traditional pattern). In conclusion, whereas the Western pattern was directly associated with colon cancer, the prudent pattern was strongly protective for rectal cancer. PMID- 21627379 TI - Association between alcohol consumption and colorectal carcinogenesis: an ecological study in Korea. AB - The sharp rise in the incidence rate of colorectal cancer (CRC) in Korea over the past decades may be partly attributed to increase in alcohol consumption. An ecological study was conducted in Korea to determine the association between alcohol consumption and the risk of colorectal cancer. We obtained the CRC incidence rates for the years 1999?2007 from the Korean Central Cancer Registry and the data on national alcohol consumption for the years 1998, 2001, 2005, and 2007 from the reports of the Korea National Health and Nutrition Examination Survey. Pearson's correlation coefficients were determined using data for alcohol intake and CRC incidence rate. People who consumed more than 45 g alcohol/day were defined as heavy drinkers. A significant correlation between alcohol consumption and the CRC incidence rate was observed in men; Pearson's correlation coefficients were statistically significant for men (r = 0.99; P = 0.001), but not for women (r = 0.82; P = 0.180). In the <50-year age group, the age-specific incidence rate for men was comparable to that for women, but in the >= 50-year age group, it increased rapidly in men. The increase in alcohol consumption appears to be attributable to increase in the number of heavy drinkers among men aged 25-59 years, particularly among men aged 45-49 years. Our findings may aid in predicting future CRC incidence in Korea. PMID- 21627380 TI - Sun protection behavior and individual risk factors of Turkish Primary School Students associated with skin cancer: a questionnaire-based study. AB - AIM: To determine the Turkish primary school students' sun protection behaviors and their individual risk factors associated with skin cancer. METHODS: The sample of this descriptive study consisted of 6th-8th graders (11-14 age group) from a primary school in the western part of Turkey. A self-report questionnaire was administered. The response rate was 86.0 %. RESULTS: The mean age of students was 12.8 years (SD =0.73). It was determined that the male students in the age group of 13-14 had more risks associated with skin cancer when compared to the female students in the same age group. In terms of the knowledge of the most dangerous hours of the sun and how to protect from their harmful effects, no significant difference was evident between females and males or across age groups. The average score of the students' frequency of use of sun protection methods is 3.40 +/- 0.67 out of 5. The most frequent methods used for sun protection were found to be seeking shade during periods of peak ultraviolet radiation and wearing light-colored clothing. The average frequency score for sun protective behavior of the 11-12 age group was significantly high (p < 0.05). CONCLUSION: It was determined that the students did not practise sun protection behavior frequently enough. PMID- 21627381 TI - Detection of high-risk human papillomavirus types 16 and 18 but not 33 and 52 in external genital warts from Iranian females. AB - BACKGROUND: External genital warts (EGW) are relatively common sexually transmitted diseases. In the majority of cases, low-risk human papilomaviruses (HPV), such as HPV-6 and HPV-11, are responsible but, high-risk types may also be detected and this has a bearing on vaccines for cervical cancer prevention. In this study the incidence of the high-risk HPV types 16, 18, 33 and 52 in EGWs of females from the southwest of Iran was assessed. METHODS: Seventy-nine women with EGWs participated in this study. Quantitative real-time PCR with gene specific primers and probes for the E6 gene of HPV-16, 18, 33 and 52, were used for the detection of HPV DNA in the tissue and blood samples. RESULTS: Of the 79 tissue specimens, 13 (16.5%) were HPV positive, only genetic materials of HPV-16 and HPV 18 being detected, twelve patients (15.2%) were positive only for HPV-18 and the coexistence of HPV-16 and HPV-18 was shown in one patient. Only one plasma sample showed evidence of HPV-16 with very low viral load. CONCLUSION: Our data showed that high-risk HPV types can be found in the tissue specimens of EGW samples obtained from female patients in the Southwest of Iran, with HPV-18 as the most abundant type; however, additional studies with a larger population are required to prove the finding and help to determine the most appropriate type of virus for vaccine design for Iranian women. PMID- 21627382 TI - Ubiquitin-specific protease 14 expression associated with intrahepatic cholangiocarcinoma cell differentiation. AB - The purpose of this study was to identify the gene alterations amplified from AO16 primer and examine whether the expression patterns of USP14 in clinical specimens from patients with intrahepatic cholangiocarcinoma (ICC) is associated with cancer cells. DNA from tumor and corresponding normal tissues of 52 patients was amplified with 33 arbitrary primers. The DNA fragment that altered most frequently in ICC was cloned, sequenced, and identified by comparison with known nucleotide sequences in the genome database. The DNA copy numbers of the allelic alterations in cholangiocarcinoma were determined by quantitative real-time PCR and interpreted as allelic loss or DNA amplification by comparison with the reference gene. Associations between allelic imbalance and clinicopathological parameters of ICC patients were evaluated by X2-tests. The Kaplan-Meier method was used to analyze survival rates. Immunohistochemically, USP14 showed weak cytoplasmic staining in normal bile duct epithelial cells. It was strongly detected in 21 cancer patients (43.8%). There were correlations between USP14 expression level and the clinicopathological features of ICC, histological grade (P < 0.05). However, there were no significant differences in age, gender, tumor size, metastasis, lymph node metastasis, and staging. USP14 expression was related to cholangiocarcinoma cell differentiation. Due to their emerging role in control of multiple signaling pathways and oncoproteins, USP14 inhibitors may be useful for anticancer agents. PMID- 21627383 TI - Life-style and genital human papillomavirus in a cross-sectional survey in Shanxi Province, China. AB - BACKGROUND: To determine the age-stratified HPV prevalence rate and the risk factors of life-style associated with HPV infection among women in rural China. METHODS: An age-stratified, cross-sectional survey of 941 women between 16-59 years old was conducted in rural China. Carcinogenic HPV infection was determined using Digene's Hybrid Capture II HPV DNA test and interviews of life-style were conducted. Odds ratios (OR) and 95% confidence interval (CI) from the logistic regression models were used to determine the risk factors associated with HPV. RESULTS: Among 941 women, 745 who had sexual intercourse underwent a cervical examination. The prevalence rate of 13 carcinogenic HPV infections among women 20 59 years old was 15.97%. the rate of HPV prevalence in the 25-34 age group was statistically lower than that in 20-24 and 35-59 age groups (X2=13.3, P=0.0013). The OR of bathing every 7-19 days, 20-180 days, less than once every 180 days vs. bathing at least once a week were 1.19, 1.83 and 2.29 respectively and they had a dose-response relationship (Trend Test: P=0.003). The OR of women aged 25-34 age group vs. 20-24 age group was 0.40 (0.16 - 0.97) and the OR of bathing once every 180 days or less vs. at least once weekly was 2.22 (1.14 - 4.33) adjusted for the other confounding factors,. CONCLUSIONS: The HPV prevalence rate was lowest among child bearing women aged 25-34 year. Also, personal hygiene is significantly associated with the HPV infection in this area, regardless of age. PMID- 21627384 TI - Physiological, reproductive factors and breast cancer risk in Jiangsu province of China. AB - To evaluate the relationship between physiological, reproductive factors and risk of breast cancer, we conducted a case-control study with 669 cases and 682 population-based controls in Jiangsu Province of China. A structured questionnaire was used to elicit detailed information. All subjects completed an in-person interview. Unconditional logistic regression analysis was performed to calculate odds ratios (ORs) and 95% confidence intervals (CIs) as measures of risk for breast cancer. The results have revealed that there was an increasing risk of breast cancer, include early age at menarche(<= 13 year), late age at menopause(< 50 year) and older age at first pregnancy (<= 30 year). Breastfeeding was associated significantly with a reduced risk of breast cancer. Women who had history of breastfeeding were at significantly decreased OR (0.44, 95%CI: 0.27 0.73). The protective effects of breastfeeding for breast cancer seemed greater for women who had extended duration of breastfeeding during their lifetime (p for linear trend: 0.0095). These results suggested that physiological and reproductive factors may play important roles in the development of breast cancer among Jiangsu' women of China. PMID- 21627385 TI - SLIT2 is epigenetically silenced in ovarian cancers and suppresses growth when activated. AB - OBJECTIVES: The SLIT2 gene is a novel tumor suppressor gene, whose hypermethylation has been detected in several malignances, including breast cancer, colorectal carcinoma and gliomas. In this study, we assessed the status of SLIT2 and its functions in ovarian cancers and cell lines. METHODS: Methylation-specific PCR was used to investigate the methylated promoter of SLIT2; the functions of SLIT2 in ovarian cancer cells were measured by MTT, colony formation assay and flow cytometry. RESULTS: SLIT2 promoter hypermethylation was detected in 56 of 66 (84.8%) ovarian cancer samples and downregulation of SLIT2 expression in 52 (78.8%). The decreased expression was significantly correlated with SLIT2 promoter hypermethylation (p<0.01). Moreover, reversed expression of SLIT2 suppressed cell growth, migration, colony formation abilities and induced more apoptosis. CONCLUSIONS: These results suggest that SLIT2 is a tumor suppressor in ovarian cancer, and may be a novel target for ovarian cancer treatment. PMID- 21627386 TI - Informational needs of breast cancer patients on chemotherapy: differences between patients' and nurses' perceptions. AB - BACKGROUND: Cancer and chemotherapy are sources of anxiety and worry for cancer patients. Information provision is therefore very important to empower them to overcome and adjust to the stressful experience. Thus, nurses should be aware of the informational needs of the patients throughout the course of their care. PURPOSE: The purpose of the study was to identify the important information required by breast cancer patients during the first and fourth cycles of chemotherapy from both the patients' and nurses' perceptions. METHODOLOGY: This is a longitudinal study used a questionnaire adapted from the Toronto Informational Needs Questionnaires-Breast Cancer (TINQ-BC). Some modifications were made to meet the specific objectives of the study. The study was conducted in the Chemotherapy Day Care at the University of Malaya Medical Centre (UMMC), Malaysia. A total of 169 breast cancer patients who met the inclusion criteria, and 39 nurses who were involved in their care were recruited into the study. RESULTS: The overall mean scores at first and fourth cycle of chemotherapy were 3.91 and 3.85 respectively: i.e., between 3 (or important) and 4 (or very important), which indicated a high level of informational needs. There was no significant difference in information needed by the breast cancer patients between the two cycles of chemotherapy (p=0.402). The most important information was from the subscale of disease, followed closely by treatment, physical care, investigative tests and psychosocial needs. Nurses had different views on the important information needed by breast cancer patients at both time points (p = 0.023). CONCLUSIONS: Breast cancer patients on chemotherapy have high levels of informational needs with no significant differences in information needed at first cycle as opposed to fourth cycle. There were differences between the perceptions of the breast cancer patients and the nurses on important information needed. A paradigm shift, with an emphasis on patients as the central focus, is needed to enhance the information giving sessions conducted by nurses based on the perceptions of the patients themselves. PMID- 21627387 TI - A C to T polymorphism of urokinase plasminogen activator (P141L) is associated with Helicobacter pylori infection. AB - Urokinase plasminogen activator (uPA) plays an important role in tumor invasion and certain inflammatory diseases. However, few studies have paid attention to how the uPA is associated with Helicobacter pylori infection and gastric atrophy. This study investigated associations of a C-to-T polymorphism of uPA (P141L, rs2227564) in exon 6 in 454 Japanese health checkup examinees (126 males and 328 females) aged 35 to 85 without a history of cancer. The uPA was genotyped by polymerase chain reaction with two-pair primers. The genotype distribution was in Hardy-Weinberg equilibrium (p=0.52) and the frequency of the T allele was 0.239. The risk of H. pylori sero-positivity was significantly reduced with the T/T genotype; the odds ratio (OR) relative to the C/C genotype was 0.34 (95% confidence interval [CI]: 0.14 to 0.86). Of the sero-negative subjects, 21 with atrophy were infected with H. pylori but lost their sero-positivity. After reclassifying them together with the sero-positive subjects, the corresponding OR was 0.40 (95% CI: 0.16 to 1.00), confirming that the T/T genotype decreased the risk of H. pylori infection. This gene polymorphism was not associated with the risk of gastric atrophy. In conclusion, this study indicated a possibility that the uPA minor homozygous genotype was associated with a reduction of H. pylori infection risk. Further studies are required to confirm these findings. PMID- 21627388 TI - Treatment of cervical carcinoma with high-dose rate intracavitary brachytherapy: two years follow-up study. AB - AIMS: This study focused on pelvic recurrence rate and late complications following treatment with high dose rate brachytherapy with a three fractionation scheme. SETTING AND DESIGN: This retrospective observational study was conducted from 1st November 2003 to 31st March 2005 at a tertiary care centre. METHODS AND MATERIALS: Women were treated with external beam radiotherapy and three fractions of high dose rate brachytherapy, divided into two broad groups IIB+ IIIA and IIIB+IVA. Duration of follow-up was 2 years and main outcome measures were recurrence and rectal and urinary bladder complications. Results were assessed with the Chi square test and P-values using an alpha level of 0.05 for Type I error. RESULTS: Of the total of 286 women, 72 (25.4%) developed central-regional recurrence. Overall two year pelvic control rate was 74.6%, with values of 78.1% and 72.8% for stages IIB+ IIIA, IIIB+IVA, respectively. Five women developed distant metastasis and 21.5% suffered low grade rectal complications. After two years the prevalence of bladder complications was only 5.4%. CONCLUSION: Using a three fraction scheme, high dose rate brachytherapy is safe and effective in the management of cervix cancer. PMID- 21627389 TI - Predictors of smoking cessation among staff in public Universities in Klang Valley, Malaysia. AB - Smoking cessation studies are often performed in clinic based settings. The present example aimed to find predictors of success among staff in worksite smoking cessation programmes in two major public universities in Klang Valley, Malaysia. All staff from both universities received an open invitation via staff e-mail and letters to participate. At the start of treatment, participants were administered the Rhode Island Stress and Coping Questionnaire and Family Support Redding's Questionnaire. Behaviour therapy with free nicotine replacement therapy (NRT) were given as treatment. After two months, they were contacted to determine their smoking status. 185 staff from University A (n=138) and University B (n=47), responded and voluntarily showed interest to quit. There was no significant difference in respondents with respect to socio demographic characteristics and smoking history. After two months of treatment, quit rates were 24% in University A vs. 38 % in University B (p>0.05). Univariate predictors of cessation were adherence to NRT (p<0.001), smoking fewer cigarettes per day (p<0.05) and the number of behaviour therapy sessions attended (p<0.001). Logistic regression identified 3 significant predictors of smoking cessation. Participants attending more than one session (OR= 27.00; 95% CI : 6.50; 111.6), and having higher pre-treatment general stress (OR= 2.15; 95% CI: 1.14; 4.05) were more likely to quit, while a higher number of cigarettes smoked (OR= 0.19: 95% CI: 0.06; 0.59) reduced the likelihood of quitting. Increasing age, ability to cope with stress and family support were not significant predictors. We conclude that factors such as the number of counseling sessions, the amount of cigarettes smoked at baseline, adherence to NRT and pretreatment stress are important considerations for success in a worksite smoking cessation programme. PMID- 21627390 TI - Risk of breast cancer, health beliefs and screening behaviour among Turkish academic women and housewives. AB - BACKGROUND: Breast cancer (BC) is one of the most common forms of epithelial neoplasms type of cancer affecting women worldwide. The main objective of this study was to examine health beliefs, screening behavior and risk for developing BC in academic women and housewives in Turkey. METHODS: In this cross-sectional, descriptive study, a sample of 415 women were surveyed, composed of 166 academics and 249 housewives between the ages of 20-65 years. RESULTS: Risk of developing breast cancer, both five-year and lifetime, was higher in academic women than in housewives (P < 0.001). The frequency of BSE, CBE and mammography was higher in academic women than housewives. For academic women, perceptions related to benefit, self-efficacy, health motivation were significantly higher than housewives (t= 3.726, P < 0.001; t=8.165, P < 0.001, t=2.326; P < 0.021, respectively). CONCLUSION: The findings of this study indicate that the risk for developing BC in academic women is higher than for housewives. Although academic women had more screening performance for early diagnosis of BC, it was still lower than 50%, below the desired level. Education programs should be aimed at increasing women's BC screening behaviors and the positively affect beliefs. PMID- 21627391 TI - Knowledge about human papillomavirus infection and cervical cancer prevention among nurses in Chiang Mai University Hospital, Thailand. AB - This study was undertaken to evaluate knowledge about HPV infection and cervical cancer among nurses in Chiang Mai University Hospital, Thailand. The 16 questions evaluating knowledge were 'true/false/do not know' type. Two hundred and twenty nurses agreed to participate in this survey. Most knew that cervical cancer is the most common female cancer in Thailand (92.7%), HPV infection is a causal factor of cervical cancer (81.8%), early stage cervical cancer is curable (94.1%), and an adequate scale of cervical screening could prevent morbidity and mortality from cervical cancer (86.8%). The majority of participants (more than 70%) correctly acknowledged risk factors for cervical cancer as smoking, having multiple sexual partners, and sex at an early age. However, the majority of participants did not know that HPV infection and early stage cervical cancer are commonly asymptomatic. In conclusion, knowledge regarding cervical cancer among nursing staff in the author's institute is considerably favorable. However, their understanding about the natural history of HPV infection and cervical cancer is suboptimal, and requires further attention if an effective cervical cancer screening program is to be implemented. PMID- 21627392 TI - Management of breast lesions by breast physicians in a heavily populated South asian developing country. AB - BACKGROUND: In Asia, from 1998 to 2002, the highest annual-age standardized incidence rates of breast cancer per 100,000 women were recorded as follows: in the Karachi South district of Pakistan 69.0 and in the Israeli Jews 96.8. At Shaukat Khanum Memorial Cancer Hospital and Research Center in Lahore, Pakistan, in 15-years from Dec. 1995-Dec. 2009, among adult females, approximately 46% (8,915) of malignancies were recorded as breast tumors. Further, according to Pakistan's population estimates (2009), the total population of the country is 177 million; females 85 million (40-69 years: 13.6 million). DISCUSSION: Screening of asymptomatic women: Basing the recommendations on biennial mammograhic screening for average-risk women in the 40-69 year age-band, about 6.8 million women will have to be screened every year. In a resource-constrained country like Pakistan, early detection by this method is not possible. As most symptomatic women present with advanced disease, clinicians skilled in breast diseases are required. The Australasian Society of Breast Physicians has developed a formal three year training model for General Practitioners to qualify as breast physicians by: i) developing their skills in the areas of clinical breast-examination, interpretation of mammography and breast ultrasound; performance of image-guided interventional procedures; counseling of and planning/coordinating treatment of females with breast cancer and assessment/monitoring of women at potentially 'high-risk' of cancer; and ii) working in consultation with surgeons, radiologists, pathologists, oncologists, and other members of the multidisciplinary team. SUMMARY: Easily accessible one stop breast clinics staffed by trained breast physicians can help reduce morbidity/mortality from breast cancer in developing countries, and improve the quality of life and survival. PMID- 21627393 TI - Use of intravenous morphine for acute decompensated heart failure in patients with and without acute coronary syndromes. AB - BACKGROUND: Current guidelines regarding the use of intravenous morphine (IM) in the management of patients with acute decompensated heart failure (ADHF) are discordant; whereas the American guidelines reserve IM for terminal patients, the European guidelines recommend its use in the early stage of treatment. Our aim was to determine the impact of IM on outcomes of ADHF patients. METHODS: Stepwise logistic regression and propensity score analysis of ADHF patients with and without use of IM was performed in a national heart failure survey. RESULTS: Of the 4102 enrolled patients, we identified 2336 ADHF patients, of whom 218 (9.3%) received IM. IM patients were more likely to have acute coronary syndromes, acute rather than exacerbation of chronic heart failure, and diabetes mellitus and dyslipidemia. They had higher heart rate, were less likely to receive diuretics and more likely to receive aspirin and statins. Unadjusted in-hospital mortality rates were 11.5% versus 5.0% for patients who did or did not receive IM, and the adjusted odds ratio (OR) for in-hospital death was: 2.0 (1.1 - 3.5, P = 0.02). Using propensity analysis, we identified 218 matched pairs of patients who did or did not receive IM. In multivariable analysis accounting for the propensity score (c-statistic 0.82), IM was not associated with increased in-hospital death (OR: 1.2 (0.6 - 2.4), P = 0.55). CONCLUSION: IM was used sparingly in our ADHF cohort, and was independently associated with increased in-hospital death in multivariable analysis, but not in propensity score analysis. Thus, IM may be used in ADHF, but with caution. Further randomized trials are warranted. PMID- 21627394 TI - Pre-hospital treatment of STEMI patients. A scientific statement of the Working Group Acute Cardiac Care of the European Society of Cardiology. AB - In ST-elevation myocardial infarction (STEMI) the pre-hospital phase is the most critical, as the administration of the most appropriate treatment in a timely manner is instrumental for mortality reduction. STEMI systems of care based on networks of medical institutions connected by an efficient emergency medical service are pivotal. The first steps are devoted to minimize the patient's delay in seeking care, rapidly dispatch a properly staffed and equipped ambulance to make the diagnosis on scene, deliver initial drug therapy and transport the patient to the most appropriate (not necessarily the closest) cardiac facility. Primary PCI is the treatment of choice, but thrombolysis followed by coronary angiography and possibly PCI is a valid alternative, according to patient's baseline risk, time from symptoms onset and primary PCI-related delay. Paramedics and nurses have an important role in pre-hospital STEMI care and their empowerment is essential to increase the effectiveness of the system. Strong cooperation between cardiologists and emergency medicine doctors is mandatory for optimal pre-hospital STEMI care. Scientific societies have an important role in guideline implementation as well as in developing quality indicators and performance measures; health care professionals must overcome existing barriers to optimal care together with political and administrative decision makers. PMID- 21627395 TI - Myocardial ischemia secondary to congenital hypoplastic left coronary cusp in adult. AB - Coronary arterial obstruction associated with congenital aortic valve disease is rare in childhood, and has not been reported in adult. Here we reported a 49-year old healthy woman with hypoplastic left coronary cusp resulting in myocardial ischemia in the territory of left main coronary artery. PMID- 21627396 TI - Evaluation of a new safety peripheral IV catheter designed to reduce mucocutaneous blood exposure. AB - OBJECTIVES: We evaluated performance and clinical acceptability of a new peripheral intravenous catheter (PIVC) designed to reduce blood exposure. METHODS: A two phased, unblinded, randomized controlled trial was conducted at a clinical research center in New Jersey, USA. In Phase 1, clinicians were asked to evaluate two devices: a PIVC with blood control (BD Insyte Autoguard * BC [Blood Control] Shielded IV Catheter), and a reference conventional PIVC (BD Insyte Autoguard Shielded IV Catheter). In Phase 2, clinicians compared two insertions of the investigational PIVC with blood control (PIVC-BC); one with venous compression and one without. The PIVC-BC was evaluated for superiority to the conventional PIVC with regard to blood exposure and for equivalence in general performance characteristics. RESULTS: Seventy-eight clinicians (mean age: 41.4 years; 89.7% female) and 234 healthy volunteers (mean age: 40.2 years; 61.5% female) were enrolled. Blood leakage occurred significantly more in the conventional PIVC group (39.1%) as compared to the PIVC-BC group (2.0%) (difference: 37.1% [95% CI: 28.8%; 45.15%]). Blood leakage rates for the PIVC-BC with or without use of venous compression were similar, 2.6% and 1.3% respectively (difference: 1.3% [95% CI: -7.8%; 4.7%]). A total of 98.7% of clinicians rated the PIVC-BC as clinically acceptable compared to 89.6% with the reference PIVC (difference: -9.1; 95% CI: -18; -1.5%) and 98.7% agreed it replaced the need for venous compression during catheter insertion (95% CI: 92.8%; 100%). Although the inability to blind clinicians to the investigational product was a potential source of bias, this was unlikely to affect assessments of observed blood leakage. CONCLUSIONS: The PIVC with blood control demonstrated reduced blood leakage during insertion and was rated no different for clinical acceptability and insertion performance compared to the conventional PIVC. Clinicians agreed that the new design replaced the need for venous compression to control blood flow during IV catheter insertion. PMID- 21627397 TI - Fundamental movement skills and self-concept of children who are overweight. AB - OBJECTIVE: Differences in fundamental movement skills and self-perceptions of physical ability and physical appearance of overweight and non-overweight children were investigated. METHOD: Overweight (n = 89, mean age = 8.75 +/- 1.4 years, BMI z-score = 2.22, SD = 0.46, 46% male) and non-overweight (n = 27, mean age = 8.25 +/- 1.5 years, BMI z-score = 0.03, SD = 0.73, 62.1% male) participants enrolled in the KOALA (Kinder Overweight Activity Lifestyle Actions) project were included. The overall objective of the KOALA project was to determine in a randomized controlled trial the effect of a Triple P (Positive Parenting Program), and a family 'Eat Well Be Active' Scouts Camp program on BMI in overweight children. Baseline between-group differences on measures of fundamental movement skills and self-concept perceptions were analyzed using independent samples t-tests. Relationships between BMI and these variables were investigated with multiple linear regression. RESULTS: Overweight children had lower scores on Bruninks-Oseretsky Test of Motor Performance-2 subtests (Bilateral Coordination, Upper Limb Coordination, Strength, Balance, and Running Speed and Agility), and Physical abilities self-concept than non-overweight children. CONCLUSIONS: Children who were overweight had significant fundamental movement skill difficulties, as well as having poorer Physical abilities self concept perceptions compared to non-overweight children. The association between increasing BMI and poor performance of gross motor tasks has potential implications for physical activity participation. Future research is needed to determine if fundamental movement skill difficulties and low physical ability self-concept are predisposing factors for children who are overweight or associated outcomes. PMID- 21627398 TI - Sequential therapy versus standard triple drug therapy for eradication of Helicobacter pylori in patients with perforated duodenal ulcer following simple closure. AB - BACKGROUND: Resistance to clarithromycin, a component of standard triple therapy, leads to inconsistent eradication rates of Helicobacter pylori infection. Some studies have shown higher eradication rates for H. pylori using sequential regimen. This study was done to compare the eradication rates for H. pylori infection between the standard triple drug therapy and the sequential therapy. METHODS: Seventy-three patients with perforated duodenal ulcer following simple closure with H. pylori infection were randomized to receive either standard triple drug therapy or the sequential therapy. Standard triple drug therapy comprised of omeprazole, clarithromycin, and amoxicillin for 10 days. Sequential therapy comprised of omeprazole and amoxicillin or the first 5 days followed by omeprazole, clarithromycin, and amoxicillin for the next 5 days. Follow-up endoscopy was done at 2 months to assess the eradication rates, compliance, and side effects with each regimen. RESULTS: Eradication rates for standard triple therapy and sequential regimen were 81.25% and 87.09%, respectively (p = 0.732). The cost of sequential therapy was cheaper and incidence of side effects and compliance were similar in each group. CONCLUSION: Standard triple therapy and sequential therapy have similar efficacy for eradication of H. pylori and sequential therapy is an economical alternative to standard triple therapy. PMID- 21627399 TI - Oral supplementation of turmeric attenuates proteinuria, transforming growth factor-beta and interleukin-8 levels in patients with overt type 2 diabetic nephropathy: a randomized, double-blind and placebo-controlled study. AB - OBJECTIVE: End-stage renal disease (ESRD) due to type 2 diabetic nephropathy is a very common condition which is increasing in prevalence, and is associated with high global levels of mortality and morbidity. Both proteinuria and transforming growth factor-beta (TGF-beta) may contribute to the development of ESRD in patients with diabetic nephropathy. Experimental studies indicate that turmeric improves diabetic nephropathy by suppressing TGF-beta. Therefore, this study investigated the effects of turmeric on serum and urinary TGF-beta, interleukin-8 (IL-8) and tumour necrosis factor-alpha (TNF-alpha), as well as proteinuria, in patients with overt type 2 diabetic nephropathy. MATERIAL AND METHODS: A randomized, double-blind and placebo-controlled study was carried out in the Diabetes Clinic of the Outpatient Department of Shiraz University of Medical Sciences on 40 patients with overt type 2 diabetic nephropathy, randomized into a trial group (n = 20) and a control group (n = 20). Each patient in the trial group received one capsule with each meal containing 500 mg turmeric, of which 22.1 mg was the active ingredient curcumin (three capsules daily) for 2 months. The control group received three capsules identical in colour and size containing starch for the same 2 months. RESULTS: Serum levels of TGF-beta and IL-8 and urinary protein excretion and IL-8 decreased significantly comparing the pre- and post-turmeric supplementation values. No adverse effects related to turmeric supplementation were observed during the trial. CONCLUSION: Short-term turmeric supplementation can attenuate proteinuria, TGF-beta and IL-8 in patients with overt type 2 diabetic nephropathy and can be administered as a safe adjuvant therapy for these patients. PMID- 21627400 TI - Treatment of female stress urinary incontinence: what women find acceptable and the impact of clinical and urodynamic evaluation on their final choice. AB - OBJECTIVES: The aim of this study was determine what women find acceptable regarding treatment modality for Stress urinary incontinence (SUI) and to assess the impact of clinical and urodynamic evaluation on their final decision. MATERIAL AND METHODS: This prospective, institutional review board-approved study included 100 consecutive women with primary SUI. All women were given a questionnaire that included the Urinary Distress Inventory (UDI-6) and the American Urologic Association Quality of Life questionnaire (AUA-QoL6). The patients were also asked to choose one of the four available treatment options, which included major surgeries, minor surgeries, office procedures and medication. Factors affecting the initial choice of treatment were studied. A Q tip test and multichannel urodynamics were then carried out and the patients were recounseled by the urologist. The patients' final decision was compared with their initial choice and statistical analysis was performed. RESULTS: Initially, 22% patients chose major surgery, 39% minor surgery, 27% an office procedure and 12% medication. This was affected by age, symptom severity and quality of life bother. After clinical and urodynamic evaluation, 34% shifted to a different treatment modality; this correlated significantly with young age, severe symptoms, limited urethral mobility and low Valsalva leak point pressure. CONCLUSIONS: Patients' initial choice for treatment of SUI was affected by age, symptom severity and quality of life; however, the final decision was more influenced by the clinical and urodynamic evaluation. It is important to counsel the patients before definitive treatment, to achieve a better outcome. PMID- 21627401 TI - Cytotoxity of nanoparticles is influenced by size, proliferation and embryonic origin of the cells used for testing. AB - Cytotoxicity screening is a common technique in drug compound screening for the identification of adverse cellular effects. Nanoparticles may cause interference in these assays. For the interpretation of cytotoxicity data it is important to study also the influence of other factors like pre-treatment of the nanoparticles, the choice of the cell culture medium and type of cell used for testing. Carboxyl polystyrene particles (CPS, 20-1000 nm) were physicochemically characterized and cytotoxicity assessed with seven screening assays in 20 cell lines, which differed in species, growth pattern, cell size, doubling time, embryonic origin and capacity for phagocytosis. Small CPS acted more cytotoxic in all cell lines, larger CPS only in phagocytic cells. Small differences in cytotoxicity were noted between the screening assays. Growth pattern and cell size determined cytotoxicity more than proliferation rate and embryonic origin of cells. Non-adherent cells, cells of mesenchymal origin and with high proliferation rate may be more susceptible to damage by nanoparticles. PMID- 21627402 TI - Nano-Ag inhibiting action potential independent glutamatergic synaptic transmission but increasing excitability in rat CA1 pyramidal neurons. AB - The aim of this study was to investigate the actions of silver nanoparticles (nano-Ag) on glutamatergic synaptic transmission and excitability in hippocampal CA1 pyramidal neurons with whole cell patch technique. The amplitude of miniature excitatory postsynaptic currents (mEPSCs) was inhibited by silver nano-particles (nano-Ag) (10-5 g/ml and 10-4 g/ml), but the amplitude and frequency of spontaneous excitatory postsynaptic currents (sEPSCs) were increased by nano-Ag treatment (10-5 g/ml and 10-4 g/ml). Furthermore, nano-Ag (10-5 g/ml and 10-4 g/ml) increased the spontaneous network activity. These results provide further insights into the underlying mechanisms responsible for the effects of nano-Ag on central nervous system (CNS). PMID- 21627404 TI - In vitro neuroprotective effects of the leaf and fruit extracts of Juglans regia L. (walnut) through enzymes linked to Alzheimer's disease and antioxidant activity. AB - Several extracts of the leaves and fruits of Juglans regia L. were assessed for their neuroprotective effects through antioxidant and anti-cholinesterase methods. Anticholinesterase activity was determined against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), the enzymes vital for Alzheimer's disease, at 50, 100 and 200 MUg ml(-1). Antioxidant activity was tested using radical scavenging activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH), N,N dimethyl-p-phenylenediamine (DMPD), superoxide (SO), nitric oxide (NO) and hydrogen peroxide (H(2)O(2)) radicals, as well as ferric ion-chelating capacity, ferric- and phosphomolybdenum-reducing antioxidant power at 500, 1000 and 2000 MUg ml(-1). Total phenol and flavonoid quantification of the extracts was calculated. The extracts scavenged DPPH radical in varying degrees; however, they did not scavenge DPMD and H(2)O(2). Only the dichloromethane and water extracts were able to quench SO (10.09 +/- 1.38%) and NO (24.09 +/- 2.19%) radicals, respectively, at low level. The extracts showed either low or no BChE inhibition and no AChE inhibition. PMID- 21627403 TI - Significance of pretreatment cardiovascular morbidity as a risk factor during treatment with parenteral oestrogen or combined androgen deprivation of 915 patients with metastasized prostate cancer: evaluation of cardiovascular events in a randomized trial. AB - OBJECTIVE: This study aimed to evaluate prognostic risk factors for cardiovascular events during treatment of metastatic prostate cancer patients with high-dose parenteral polyoestradiol phosphate (PEP, Estradurin(r)) or combined androgen deprivation (CAD) with special emphasis on pretreatment cardiovascular disease. MATERIAL AND METHODS: Nine-hundred and fifteen patients with T0-4, Nx, M1, G1-3, hormone- naive prostate cancer were randomized to treatment with PEP 240 mg i.m. twice a month for 2 months and thereafter monthly, or to flutamide (Eulexin(r)) 250 mg per os three times daily in combination with either triptorelin (Decapeptyl(r)) 3.75 mg i.m. per month or on an optional basis with bilateral orchidectomy. Pretreatment cardiovascular morbidity was recorded and cardiovascular events during treatment were assessed by an experienced cardiologist. A multivariate analysis was done using logistic regression. RESULTS: There was a significant increase in cardiovascular events during treatment with PEP in patients with previous ischaemic heart disease (p = 0.008), ischaemic cerebral disease (p = 0.002), intermittent claudication (p = 0.031) and especially when the whole group of patients with pretreatment cardiovascular diseases was analysed together (p < 0.001). In this group 33% of the patients had a cardiovascular event during PEP treatment. In the multivariate analysis PEP stood out as the most important risk factor for cardiac complications (p = 0.029). Even in the CAD group there was a significant increase in cardiovascular events in the group with all previous cardiovascular diseases taken together (p = 0.036). CONCLUSIONS: Patients with previous cardiovascular disease are at considerable risk of cardiovascular events during treatment with high-dose PEP and even during CAD therapy. Patients without pretreatment cardiovascular morbidity have a moderate cardiovascular risk during PEP treatment and could be considered for this treatment if the advantages of this therapy, e.g. avoidance of osteopenia and hot flushes and the low price, are given priority. PMID- 21627405 TI - Yoga for asthma? A systematic review of randomized clinical trials. AB - OBJECTIVE: The objective of this systematic review was to assess the effectiveness of yoga as a treatment option for asthma. METHOD: Seven databases were searched from their inception to October 2010. Randomized clinical trials (RCTs) and non-randomized clinical trials (NRCTs) were considered, if they investigated any type of yoga in patients with asthma. The selection of studies, data extraction, and validation were performed independently by two reviewers. RESULTS: Six RCTs and one NRCT met the inclusion criteria. Their methodological quality was mostly poor. Three RCTs and one NRCT suggested that yoga leads to a significantly greater reduction in spirometric measures, airway hyperresponsivity, dose of histamine needed to provoke a 20% reduction in forced expiratory volume in the first second, weekly number of asthma attacks, and need for drug treatment. Three RCTs showed no positive effects compared to various control interventions. CONCLUSIONS: The belief that yoga alleviates asthma is not supported by sound evidence. Further, more rigorous trials are warranted. PMID- 21627406 TI - Effect of high-protein or normal-protein diet on weight loss, body composition, hormone, and metabolic profile in southern Brazilian women with polycystic ovary syndrome: a randomized study. AB - The aim of the present study was to assess the effects of a high protein (HP) and a normal protein (NP) diet on patients with polycystic ovary syndrome (PCOS) and body mass index-matched controls in a sample of southern Brazilian women. This 8 week randomized trial was carried out at a university gynecological endocrinology clinic and included 18 patients with PCOS and 22 controls. Changes in weight, body composition, hormone, and metabolic profile were analyzed in women randomized to receive HP (30% protein, 40% carbohydrate, and 30% lipid) or NP (15% protein, 55% carbohydrate, and 30% lipid). The energy content was estimated for each participant at 20-25 kcal/kg current weight/day. Physical activity, blood pressure, homeostasis model assessment (HOMA) index, and fasting and 2-h glucose and insulin remained stable during the intervention in PCOS and controls, even in the presence of weight loss. There were no changes in lipid profile in either group. In contrast, body weight, body mass index (BMI), waist circumference, percent of body fat, and sum of trunk skinfolds decreased significantly after both diets in both groups. Total testosterone also decreased in PCOS and controls regardless of diet. In conclusion, calorie reduction, rather than protein content, seemed to affect body composition and hormonal profile in this short-term study. These findings emphasize the role of non-pharmacological interventions to reduce weight and ameliorate the anthropometric and clinical phenotype in PCOS. PMID- 21627407 TI - Adverse perinatal outcome and maternal risk factors in population versus customized defined SGA babies. AB - OBJECTIVE: The purpose of this study was to compare population and customized based birth weight centiles in their association with perinatal outcome and maternal risk factors, in nulliparous Caucasian women in a socio-economic disadvantaged region. METHODS: We analyzed perinatal outcomes in births of 302 Caucasian women of which 155 were small for gestational age (SGA) and 147 were appropriate for gestational age (AGA). Out of the overall study group, two cohort studies were designed. One was classified by population centiles as either SGA (n= 133) or AGA (n = 169) and the other was classified by customized centiles as either SGA (n = 131) or AGA (n = 172). Maternal risk factors and operative delivery rates for fetal distress, Apgar scores, need for resuscitation and neonatal nursery care given, were determined for both customized and population based SGA babies. RESULTS: The customized SGA only group showed more mental health problems and special nursery in comparison with the AGA group. The population SGA only group had more smoking and mental health problems than the AGA group, but no differences on neonatal outcome measures. CONCLUSION: Use of customized centiles does identify an additional group neonates with a significantly higher need for special nursery admission in a homogeneous ethnic Caucasian group. PMID- 21627408 TI - Acute myocardial infarction triggered by acute pharyngitis in a patient with normal coronary arteries: what is the role of platelets? PMID- 21627409 TI - Analysis of biological properties of selected elements of haemostasis after treatment with the oxidized form of homocysteine in vitro. AB - The elevated level of homocysteine (Hcys; hyperhomocysteinemia, in relation to the total plasma Hcys concentration, >15 uM) is associated with different diseases in human, including cardiovascular diseases. In plasma, Hcys occurs in various forms (the reduced Hcys, the oxidized Hcys, homocysteine thiolactone (HTL) and a component of proteins as a result of N- or S-homocysteinylation). The mechanisms by which hyperhomocysteinemia contributes to changes of haemostasis are complex and unclear. The role of different forms of Hcys, which may be involved in the modulation of haemostatic process during hyperhomocysteinemia is also not yet well-known. Our previous works have shown that both Hcys in the reduced form and the most reactive form of Hcys-its thiolactone may modify fibrinolysis, coagulation process and biological activity of blood platelets. The mechanism by which the oxidized Hcys exerts the prothrombotic effect and influences on blood platelets or plasma remains unclear. The aim of our study in vitro was to establish and compare the influence of the oxidized Hcys (at final doses of 0.01-1 mM), the reduced Hcys (at final doses of 0.01-1 mM) and HTL (at final doses of 0.1-1 uM) on selected haemostatic properties of blood platelets (platelet aggregation and platelet microparticle formation measured by flow cytometry) and plasma (fibrin polymerization and lysis). Here, our results indicate that the oxidized Hcys, like the reduced Hcys or HTL-augmented blood platelet aggregation, stimulated polymerization of fibrinogen and reduced the fibrin lysis in plasma. But, we suggest that the most reactive form of Hcys may be HTL (at lower concentrations than Hcys) during hyperhomocysteinemia-induced cardiovascular diseases. PMID- 21627410 TI - Diurnal variation in platelet inhibition by clopidogrel. AB - Morning increase in the occurrence of myocardial infarction, stroke and sudden cardiac death is a well-recognized phenomenon, which is in line with a morning enhancement of platelet aggregation. We investigated whether platelet inhibition during clopidogrel and aspirin therapy varies during the day. Fifty-nine consecutive patients (45 men and 14 women) with first ST-segment elevation myocardial infarction (STEMI) treated with primary percutaneous coronary interventions (pPCI) on dual antiplatelet therapy were prospectively enrolled into the study. Blood samples were collected 4 days after start of clopidogrel treatment at 6.00 a.m., 10.00 a.m., 2.00 p.m. and 7.00 p.m. Arachidonic acid and adenosine diphosphate (ADP)-induced platelet aggregation were assessed by impedance aggregometry. Platelet inhibition by clopidogrel was lowest in the midmorning: median ADP-induced platelet aggregation was 55%, 17% and 27% higher at 10.00 a.m. compared to 6.00 a.m., 2.00 p.m. and 7.00 p.m., respectively (p < 0.002). Nonresponsiveness to clopidogrel defined according to the device manufacturer was 2.4-fold more frequent in the midmorning than in the early morning. We observed a more pronounced midmorning increase in ADP-induced platelet aggregation in diabetic patients when compared to non-diabetics. In contrast, no diurnal variation in the antiplatelet effect of aspirin was observed. In conclusion, in patients presenting with STEMI undergoing pPCI, platelet inhibition by clopidogrel is less strong in the midmorning hours. This periodicity in platelet aggregation in patients on dual antiplatelet therapy should be taken into consideration when assessing platelet function in clinical studies. PMID- 21627411 TI - Genetic determinants of on-clopidogrel high platelet reactivity. AB - Clopidogrel has been used (alone or in association with aspirin) to prevent vascular complications in atherothrombotic patients, to prevent stent thrombosis (ST) in patients undergoing percutaneous coronary intervention (PCI) and as a long-term prevention of cardiovascular and cerebrovascular events. Unfortunately, it is important to note that there are a number of patients who, during clopidogrel therapy, show and maintain a high platelet reactivity (PR), similar to that observed before the start of antiplatelet therapy. Clopidogrel pro-drug is absorbed in the intestine and this process is influenced by P-glycoprotein-1 (P-GP). Its conversion into 2-oxo clopidogrel is regulated by cytochromes (CYP) called CYP2C19, CYP2B6 and CYP1A2. Whereas, the final transformation into the active metabolite is regulated by CYP called CYP2C19, CYP2C9, CYP2B6, CYP3A4, CYP3A5 and, as recently emerged, by the glycoprotein paraoxonase-1 (PON1). The genes encoding these enzymes are characterized by several polymorphisms. Some of these are able to modify the activity of proteins, reducing the concentration of active metabolite and the values of on-clopidogrel PR. Only one gene polymorphism (CYP2C19*17) increases the clopidogrel metabolization and so the clopidogrel induced platelet inhibition. Several studies have clearly associated these gene polymorphisms to both ischemic and bleeding complications in patients receiving dual antiplatelet therapy. The aim of this review is to describe the principal gene polymorphisms influencing on-clopidogrel PR and their relationship with long term clinical outcome. PMID- 21627412 TI - Immunotherapy against invasive fungal diseases in stem cell transplant recipients. AB - Despite the availability of new antifungal compounds, morbidity and mortality of invasive fungal disease in allogeneic hematopoietic stem cell recipients are still unacceptably high. Over the past decade, one could witness an exciting improvement of the understanding of the molecular pathogenesis and of the complexity of host antifungal immune responses. This, in turn, provides critical information to augment host immunity against fungal pathogens. Strategies for enhancing the immune system include the administration of effector and regulatory cells (e.g., granulocytes, antigen-specific T cells, dendritic cells) as well as the administration of recombinant cytokines, interferons and growth factors (e.g., interferon-gamma, keratinocyte growth factor, granulocyte- and granulocyte macrophage colony stimulating factor). One has to recognize at the same time, however, that data of in vitro assays and animal models cannot necessarily be transferred into the clinical setting. In addition, meaningful clinical trials in allogeneic stem cell recipients suffering from invasive fungal disease require sufficiently large and homogenous cohorts of patients and can only be performed in international collaboration, but may ultimately improve the outcome of allogeneic transplant recipients with invasive fungal disease. PMID- 21627413 TI - Uterine scar defects. PMID- 21627414 TI - The sacrospinous ligament: conveniently effective or effectively convenient? AB - The sacrospinous ligament has been used for over 50 years as a convenient structure for treating vaginal vault and more recently, uterine prolapse. The procedure has evolved over the years and its efficacy has been hotly debated with invariable comparisons made to abdominal sacral colpopexy. Mesh surgery has introduced a newer dimension to the debate. This review is an attempt to clarify the anatomy, reflect on various techniques and offer a critique on the current 'status' of the sacrospinous ligament. PMID- 21627415 TI - Pregnancy after rupture of the pregnant uterus. AB - Pregnant women managed by us after suture repairs for rupture of the pregnant uterus between 2006 and 2009 in Enugu, South-east Nigeria were reviewed. Five pregnancies were managed during the study period. Four had normal pregnancies. Three booked early in pregnancy and had repeat caesarean deliveries at 36 weeks' gestation. One booked at another private hospital but had emergency caesarean section at 38 weeks' gestation. The fourth woman had repeat rupture at 28 weeks' gestation, suture repair and bilateral tubal ligation. Four healthy babies were discharged home. There was no maternal death. The outcomes of pregnancies after rupture of the pregnant uterus are good. Suture repairs should be the method of choice whenever preservation of future fertility is necessary. The woman and her family must be counselled on the associated risks and the need for hospital delivery by elective caesarean section in the subsequent pregnancies before discharge. PMID- 21627416 TI - FlowCytomix analysis for Toxoplasma gondii infection in pregnant women in central Taiwan. AB - The study was to determine the seroprevalence of toxoplasmosis in the sera of pregnant women in central Taiwan and to investigate the levels of cytokine in the sera of pregnant women with Toxoplasma gondii infection. The 220 blood samples were collected from pregnant women. The haematological parameters of peripheral blood were analysed by a haematology analyser. Serum samples of the pregnant women were analysed by a commercially available anti-T. gondii IgM/IgG antibody enzyme-linked immunosorbent assay (ELISA) kit and FlowCytomix assays. Six (2.7%) of the sera samples had IgM anti-T. gondii antibodies, and twenty (9.1%) had T. gondii IgG seropositive. All six IgM seropositive samples had low IgG avidity, indicative of acute infection. Total white blood cells and eosinophils were statistically significantly increased (p<0.05) in pregnant women with T. gondii infection, as compared with healthy pregnant women. Th1 cytokines IFN-gamma, IL 1beta, IL-2 and IL-12 p70, and Th2 cytokines IL-10 in pregnant women with T. gondii IgM/IgG seropositive were significantly increased (p<0.05), as compared with healthy pregnant women. These results showed that both of Th1 and Th2 cytokines play an important role in the toxoplasmosis of pregnant women. PMID- 21627417 TI - Is the use of symphysis-fundal height measurement and ultrasound examination effective in detecting small or large fetuses? AB - We investigated the accuracy of using symphysis pubis-fundal height measurement and ultrasonically derived estimation of fetal weight for identifying small-for gestational-age (SGA) and large-for-gestational-age (LGA) fetuses. A retrospective cohort study was performed using computerised records of all non diabetic women referred for an ultrasound growth scan (US) with clinically suspected SGA or LGA singleton fetuses from 35 weeks' gestation between October 2008 and March 2009 (n = 185). Birth weight data were also collected for all births over the study period (n = 3200). One-third (34%) of ultrasound estimated fetal weights were inaccurate by >10%. However, an inaccurate ultrasound estimation did not significantly increase the likelihood of spontaneous or induced onset of labour or delivery by caesarean section. Most (79%) growth scans were performed on well-grown fetuses. The majority (80%) of SGA and LGA babies in our cohort were not identified by clinicians during routine antenatal care. From 3,200 live births, there were a total of 59 infants weighing <2,500 g or >5,000 g. Of these, only 12 had been referred for an ultrasound growth scan, indicating that abdominal palpation and fundal height measurement has a 20% sensitivity in detecting SGA or LGA fetuses. Of the 12, four were detected using ultrasound, indicating a 33% detection rate. Although ultrasound has a slightly higher sensitivity, neither clinical examination using fundal height measurements nor 3rd trimester ultrasound examinations are effective at detecting SGA or LGA fetuses. PMID- 21627418 TI - Risk factors for early onset severe pre-eclampsia and eclampsia among north Indian women. AB - A case-control study was conducted in India between April 2007 and January 2008, to identify the clinical and historical risk factors associated with early onset pre-eclampsia/eclampsia (PE-E) in women attending a tertiary care hospital in North India. The study group comprised 100 women with early onset severe pre eclampsia/eclampsia (<=34 weeks) and a control group of 100 women with mild non proteinuric hypertension (>34 weeks). A detailed history including past, personal and family history, pregnancy outcome including delivery details and perinatal outcome and available investigations were recorded in a pre-designed proforma. Multiple logistic regression analysis was used to determine the risk factors for pre-eclampsia. The risk factors that were associated with increased risk of early onset severe PE-E were: history of PE-E in a previous pregnancy (adjusted odds ratio, aOR 71.40); exposure to passive smoking (aOR 16.40); inadequate antenatal supervision (aOR 15.21); family history of hypertension in one or more 1st-degree relative (aOR 8.92); living in a joint family (aOR 6.93); overweight (>120% to 150% of pre-pregnancy ideal body weight, aOR 4.65) and lower socioeconomic class (Kuppuswamy's class III-V) (aOR 3.00). Based on the above risk factors, a risk model can be constituted as practised in other places and implemented in the primary preventive measure of early-onset severe pre-eclampsia among the North Indian women attending this tertiary care hospital. PMID- 21627419 TI - Retrospective investigation of sudden maternal weight loss as an indicator of impending parturition in the term gravida: a pilot study. AB - The aim of the study was to perform a retrospective review of records to test a possible significant association between sudden, modest weight loss (<=2 lb) between the last two prenatal appointments in the late 3rd trimester and onset of labour. Hospital and clinic records were searched for women who delivered a term baby after the onset of natural labour between 2006 and 2008, in Tulsa, OK. High risk pregnancies were excluded. Patients were categorised by whether they lost or gained weight between their last two prenatal appointments. The groups' average times to onset of spontaneous labour were compared. A total of 149 records were obtained. Mothers who lost weight had an average onset of spontaneous labour in 2.30 days; their counterparts' average was 2.34 days. Student's t-test and Wilcoxon test p values were 0.95 and 0.61, respectively. A Kolmogorov-Smirnov test p value was 0.2139. A one-half standard deviation difference in time to delivery, approximately 2.5 days, was detectable with 0.95 probability. It was concluded that these data may be the first time this question has been investigated and provide evidence that sudden, modest weight loss at the end of pregnancy is not associated with shorter time to onset of natural labour. PMID- 21627420 TI - Amniotic fluid index at admission in labour as predictor of intrapartum fetal status. AB - To re-examine the value of amniotic fluid index at admission in labour (aAFI) as a predictor of intrapartum fetal status, in 326 term pregnancies at their admission in labour, labour admission test (LAT) and antepartum risk status were recorded. The different categories were correlated with fetal/neonatal outcome. There were 68.7% women at low risk on the Minnesota scale for antepartum status. The proportion of women with low aAFI (<=5 cm) was higher in the high-risk group (8 of 12, 64%, p = 0.0001) and occurrence of significantly higher non-reassuring fetal status (NRFS; 10 of 12 with low AFI, 83%, p = 0.0001). Number recording LAT as non-reactive trace (41 cases) was similar irrespective of the pregnancy status (24 and 17 in low- and high-risk cases, respectively) and fetal status through labour (21 and 20 in fetuses with NRFS and otherwise, respectively). Negative predictive value was similar between the groups (88% for aAFI and 89% for LAT), and LAT was found to be more specific (91%) than 64% for aAFI. Sensitivity and positive predictive values were poor in both. It is concluded that aAFI is not a reliable screening test to predict intrapartum fetal compromise. PMID- 21627421 TI - Maternal arrhythmias detected with electrocardiography during labour: are they significant clinically? AB - This study examines the maternal and fetal effects of arrhythmias detected by electrocardiographic (ECG) monitoring during labour in parturients at term, with no cardiovascular pathology. Pregnant cases were classified into three groups based on determined stages of labour and a standard 12-lead surface electrocardiogram and long-lead 2 rhythm strips were recorded during the labour. Cardiac arrhythmia of any kind was detected in 82.3% of patients in all stages of labour. Sinus tachycardia was the most commonly observed arrhythmia. Arrhythmias in the form of supraventricular tachycardia, T-wave inversion and ventricular extrasystole were also detected. The highest rate of arrhythmia was recorded for the active phase and 2nd phase of labour. Arrhythmias that are detected by ECG during or after the labour in patients with no cardiovascular pathology display a benign nature and do not create any clinical risk for the mother and the baby. PMID- 21627422 TI - Preoperative vaginal preparation with povidone-iodine on post-caesarean infectious morbidity. AB - The commonest complication associated with caesarean section is infection. The aim of this study is to investigate the effect of vaginal preparation with povidone-iodine on post-caesarean infection. In this clinical trial, 568 patients were selected for two groups: a treatment group and a control group, each with 284 patients. A vaginal scrub was performed before the routine abdominal scrub, with two 4 * 4 cm sponge sticks saturated with povidone-iodine solution, rotated in the vagina for about 30 s. In the control group, only the abdominal scrub was performed. Patients received a single dose of prophylactic antibiotics, and were reviewed for 6 weeks to look for predefined variables. Post-caesarean endometritis occurred less frequently in the treatment group than in the control group (2.5% vs 1.4%). There was no significant difference for febrile morbidity and wound infection in the two groups. The adjusted odds ratio for endometritis after vaginal preparation was 0.03 (95% CI: 0.008-0.7). Vaginal preparation with povidone-iodine may decrease the risk of post-caesarean endometritis. PMID- 21627423 TI - Pregnancy outcome among grand multiparous women at the University of Maiduguri Teaching Hospital: a case control study. AB - Grand multiparity has been described as an independent risk factor for a variety of obstetric complications, especially in developing countries with inadequate health facilities. This case control study compares the pregnancy outcomes of grand multiparas with that of multiparas at the University of Maiduguri Teaching Hospital over a period of 1 year. The labour ward records and patients' case notes were used to extract information. Multivariate analysis created a model of the adverse factors that were independently associated with grand multiparity after control for confounding effects of age and other variables. During the period of study, there were 1,865 deliveries, out of which 350 were grandmultiparas, a rate of 18.8%. Out of the 350 grandmultiparas, 91 were excluded, leaving 259 (74%) patients which were used for the study. The multivariate analysis shows that compared with the multiparas, the grand multiparas were more likely to be uneducated, have abruptio placentae, precipitate labour and stillbirth but were less likely to have prolonged labour, episiotomy, placenta praevia, require operative deliveries or be anaemic at booking. Grandmultiparity was found to be associated with adverse pregnancy outcome. This finding has implications for practitioners caring for these women in our environment. PMID- 21627424 TI - Obstetric outcome following free maternal care at Enugu State University Teaching Hospital (ESUTH), Parklane, Enugu, South-eastern Nigeria. AB - This study aims to determine the impact of free maternal care on the utilisation of the available delivery services and to evaluate the obstetric outcome. All deliveries at ESUTH, Parklane within the 4 months of free maternal care from 1 September to 31( )December 2008 were studied and compared with deliveries that took place 4 months before and after the free services. The results were analysed using Epi-info statistical software version 3:2:2. There was an 88% rise in the number of deliveries with the introduction of free maternal services and a 30% drop within 4 months of its termination. The maternal mortality ratio (MMR) dropped slightly, but morbidity increased significantly, as well as stillbirth rate (77/1,000 births), especially intrapartum stillbirth. Cost barrier limits women's access to healthcare in developing countries and must be addressed if we aim to achieve Millennium Development Goals (MDGs) 4 and 5. PMID- 21627425 TI - Sonographically detected caesarean section scar defects and menstrual irregularity. AB - The purpose of this study was to estimate the frequency of post-caesarean section scar defects (PCSD) on transvaginal ultrasound and to correlate these with the presence of abnormal menstrual bleeding. We reviewed the pelvic ultrasound images of 318 women referred for a gynaecological scan in a private women's ultrasound clinic. The incidence of defects at the expected location of a caesarean scar was evaluated and then correlated with patient responses to a questionnaire on abnormal menstrual bleeding. Of the 318 patients, 71 had a history of caesarean section and technically adequate pelvic ultrasound images. Fluid-filled defects in the hysterotomy incision were noted in 29 (40%). The presence of a PCSD was significantly associated with patients reporting prolonged periods or post menstrual spotting. The larger the defect, the higher was the incidence of abnormal vaginal bleeding. PMID- 21627426 TI - Symmetric dimethylarginine (SDMA) is raised in women with polycystic ovary syndrome: a pilot study. AB - Polycystic ovary syndrome (PCOS) is associated with endothelial dysfunction, which may be caused by elevated levels of asymmetric dimethylarginine (ADMA). ADMA reduces nitric oxide production in diabetes mellitus, hypertension and renal failure. Symmetric dimethylarginine (SDMA) is a stereoisomer produced alongside ADMA, and has recently been described as a risk factor for cardiovascular events. In this cross-sectional study based in a teaching hospital, 16 women with PCOS were recruited alongside 15 healthy controls, and fasting venous blood samples were obtained. Renal function was measured, and ADMA and SDMA were analysed using a high-performance liquid chromatography method. After controlling for BMI, mean ADMA and SDMA levels in women with PCOS were higher than in controls (p = 0.036 and p = 0.030, respectively). Renal function was not different between the two groups (p = 0.152). Women with PCOS have raised levels of SDMA, a molecule implicated in endothelial dysfunction and long-term cardiovascular risk. PMID- 21627427 TI - Factors affecting pregnancy outcome of intrauterine insemination cycles in couples with favourable female characteristics. AB - The aim of the presented study is to determine the effect of different sperm parameters on the pregnancy rate of intrauterine insemination (IUI) cycles in women with favourable fertility characteristics treated for infertility. Medical records of 212 infertile couples who had undergone a total of 253 cycles were reviewed retrospectively. Inclusion criteria for women were age <35 years, antral follicle count >5, FSH <15 IU/ml, and at least one patent tube documented by HSG or laparoscopy. Clinical pregnancy rates were achieved as 15.8% per cycle, and 18.8% per couple. Woman's age, partner's age, total number of motile sperm (TMS) and motility, significantly influenced pregnancy rate. Pregnancy rate was the highest when women were aged <25 and TMS >10 * 10(6). Partner's age significantly affected the pregnancy rate per cycle in women aged <30 years and TMS >10 * 10(6). Woman's age (OR: 5.4 95% CI: 1.2-24.3) and TMS (OR: 0.06 95% CI: 0.003 0.89) were predictor variables as regards to pregnancy. Pregnancy rate was the highest in IUI cycles when woman was <25 years old, TMS was >10 * 10(6), and morphology was >4%. Male age was found to be another determining factor for IUI success, even if they had a normal spermiogram. PMID- 21627428 TI - Voiding dysfunction following suburethral tape. AB - Voiding dysfunction (VD) is relatively common following suburethral tape insertion. Our study aimed to identify perioperative variables that predict VD. Women who underwent suburethral tapes (TVT(TM) and TVT-O(TM)), either as sole procedure or with a concomitant prolapse repair, were studied retrospectively. The primary outcome was women requiring catheterisation and/or re-catheterisation in the postoperative period. A total of 319 women underwent suburethral tapes within the study period: 256 case notes (80.2%) were available for review and 40/256 women (15.6%) developed postoperative VD. No preoperative urinary symptoms were associated with postoperative VD. Univariate analysis demonstrated three variables associated with VD: average flow rate (Q-ave) <=5th centile (odds ratio (OR) 2.3, 95% CI 1.2-6.5, p = 0.016), a combination of Q-ave and maximum flow rate (Q-max) <=5th centile (OR 2.8, 95% CI 1.1-6.9, p = 0.030) and concomitant prolapse procedure (OR 3.6, 95% CI 1.5-8.9, p = 0.005). Following multivariate logistic regression Q-ave <=5th centile and concomitant prolapse procedure showed the strongest association with VD. PMID- 21627429 TI - A randomised controlled trial evaluating the use of polyglactin (Vicryl) mesh, polydioxanone (PDS) or polyglactin (Vicryl) sutures for pelvic organ prolapse surgery: outcomes at 2 years. AB - The effects at 2 years of polyglactin (Vicryl) mesh inlay and polydioxanone (PDS) or polyglactin (Vicryl) suture material on prolapse symptoms, urinary, bowel, sexual function and prolapse related Quality-of-Life (QoL) in women undergoing pelvic organ prolapse surgery were evaluated in a randomised controlled trial with a 2 * 2 factorial design of Vicryl mesh (n = 32) or not (n = 34) and PDS (n = 33) or Vicryl suture (n = 33). The response rate at 2 years was 82%. There were no differences in the prolapse symptom scores between the randomised groups. Prolapse-related QoL score (mean difference: 2.05, 95% CI 0.19-3.91) and urinary incontinence score (mean difference: 2.56, 95% CI 0.02-5.11) were significantly lower (better) in women who had Vicryl compared with PDS sutures. The apparent superiority of the prolapse-related QoL and urinary incontinence scores in women using Vicryl suture material (vs PDS) needs to be confirmed in a larger trial. PMID- 21627430 TI - Sexual behaviour and HIV sero-discordance among HIV patients receiving HAART in rural Uganda. AB - Globally, HIV sero-discordance is a major public health problem, due to increased risk of HIV transmission to HIV-negative partners. From January to August 2009, we assessed the sexual behaviour of HIV-infected patients attending an HIV treatment centre in Uganda and analysed characteristics associated with HIV sero discordance. Of the 278 respondents, 256 (92.1%) were female, 218 (78.1%) were on antiretroviral drugs and 107 (38.7%) belonged to a sero-discordant couple. In the prior 3 months, 238 (88.5%) had more than one sexual partner and 98 (35.8%) had acquired new sexual partners. Only 171 (61.5%) had ever disclosed sero-status to main sexual partners. Participants who had changed sexual partners in the previous 3 months were more likely to be sero-discordant (OR 1.87, 95% CI 1.13, 3.11). Respondents who used condoms during their last sexual intercourse and those currently on ARVs were significantly less likely to be sero-discordant (OR 0.52 95% CI 0.28, 0.96 and OR 0.52 95% CI 0.30, 0.94, respectively). Only disclosure of sero-status was independently though inversely associated with discordance (adjusted OR 0.41, 95% CI 0.20, 0.64). Our findings indicate that HIV patients continue exhibiting high risk behaviour characterised by changing sexual partners, having multiple sexual partners and non-use of condoms. PMID- 21627431 TI - Rectocoele, but not cystocoele, may predict unsuccessful pessary fitting. PMID- 21627432 TI - Swine flu mimicking acute abdomen in pregnancy. PMID- 21627433 TI - Pre-labour spontaneous uterine rupture at 31 weeks' gestation following laparoscopic excision of a broad ligament paramesonephric cyst and literature review. PMID- 21627434 TI - Pregnancy after thermal balloon ablation. PMID- 21627435 TI - Twin-twin confusion syndrome: blood chimerism in opposite sex dizygotic twins. PMID- 21627436 TI - Management of fetal ovarian cysts. PMID- 21627437 TI - Alagille syndrome. PMID- 21627438 TI - Pulmonary parenchymal endometriosis presenting as bilateral pneumothoraces. PMID- 21627439 TI - Structural and functional failure of TVT by trimming of the extruded edge. PMID- 21627440 TI - Pudendal neuralgia: a rare cause of pain after tension free vaginal tape. PMID- 21627441 TI - Primary adenocarcinoma of the fallopian tube presenting as hydrosalpinx. PMID- 21627442 TI - Malignant mixed Mullerian tumour of the ovary with probable prognostic factors. PMID- 21627448 TI - Effects of low-dose gamma-rays on the embryonic development of mouse melanoblasts and melanocytes in the epidermis and hair bulbs. AB - The effects of low-dose gamma-rays on the embryonic development of animal cells are not well studied. The mouse melanocyte is a good model to study the effects of low-dose gamma-rays on the development of animal cells, as it possesses visible pigment (melanin) as a differentiation marker. The aim of this study is to investigate in detail the effects of low-dose gamma-rays on embryonic development of mouse melanoblasts and melanocytes in the epidermis and hair bulbs at cellular level. Pregnant females of C57BL/10J mice at nine days of gestation were whole-body irradiated with a single acute dose of gammarays (0.1, 0.25, 0.5, and 0.75 Gy), and the effects of gamma-rays were studied by scoring changes in the development of epidermal melanoblasts and melanocytes, hair follicles, and hair bulb melanocytes at 18 days in gestation. The number of epidermal melanoblasts and melanocytes, hair follicles, and hair bulb melanocytes in the dorsal and ventral skins was markedly decreased even at 0.1 Gy-treated embryos (P < 0.001), and gradually decreased as dose increased. The effects on the ventral skin were greater than those on the dorsal skin. The dramatic reduction in the number of melanocytes compared to melanoblasts was observed in the ventral skin, but not in the dorsal skin. These results suggest that low-dose gamma-rays provoke the death of melanoblasts and melanocytes, or inhibit the proliferation and differentiation of melanoblasts and melanocytes, even at the low dose. PMID- 21627449 TI - Zili antagonizes Bmp signaling to regulate dorsal-ventral patterning during zebrafish early embryogenesis. AB - Bone morphogenetic protein (Bmp) signaling plays a pivotal role in dorsal-ventral (DV) patterning in vertebrate embryos. Piwi proteins are required for germline and stem cell development. Our previous study demonstrated that Zili, zebrafish Piwil2, inhibits transforming growth factor (TGF)-betasignaling by interacting with Smad4, suggesting a role for zili in Bmp signaling. In the present study, zili-MO or zili mRNA was microinjected into one-cell embryos to knock down or elevate the expression of zili to study the role of zili during early zebrafish embryogenesis. Knockdown of zili inhibited the expression of dorsal marker genes, and enhanced that of ventral marker genes. In contrast, overexpression of zili promoted expression of dorsal marker genes, while it inhibited ventral marker genes. These results suggest that zili regulates DV patterning. The influence of zili on the Bmp pathway was further explored. Knockdown of zili resulted in higher expression levels of bmp2b, and bmp4, the Bmp signaling ligands, and reduced expression of chordin (chd), noggin (nog1), and follistatin (fst), which encode BMP antagonists. Meanwhile, overexpression of zili produced opposite effects. In conclusion, our results indicate that zili regulates dorsal-ventral patterning by antagonizing Bmp signaling during early embryogenesis in zebrafish. PMID- 21627450 TI - Color-pattern analysis of eyespots in butterfly wings: a critical examination of morphogen gradient models. AB - Butterfly wing color patterns consist of many color-pattern elements such as eyespots. It is believed that eyespot patterns are determined by a concentration gradient of a single morphogen species released by diffusion from the prospective eyespot focus in conjunction with multiple thresholds in signal-receiving cells. As alternatives to this single-morphogen model, more flexible multiple-morphogen model and induction model can be proposed. However, the relevance of these conceptual models to actual eyespots has not been examined systematically. Here, representative eyespots from nymphalid butterflies were analyzed morphologically to determine if they are consistent with these models. Measurement of ring widths of serial eyespots from a single wing surface showed that the proportion of each ring in an eyespot is quite different among homologous rings of serial eyespots of different sizes. In asymmetric eyespots, each ring is distorted to varying degrees. In extreme cases, only a portion of rings is expressed remotely from the focus. Similarly, there are many eyespots where only certain rings are deleted, added, or expanded. In an unusual case, the central area of an eyespot is composed of multiple "miniature eyespots," but the overall macroscopic eyespot structure is maintained. These results indicate that each eyespot ring has independence and flexibility to a certain degree, which is less consistent with the single-morphogen model. Considering a "periodic eyespot", which has repeats of a set of rings, damage-induced eyespots in mutants, and a scale-size distribution pattern in an eyespot, the induction model is the least incompatible with the actual eyespot diversity. PMID- 21627451 TI - Daytime habitat selection by introduced eastern cottontail Sylvilagus floridanus and native european hare Lepus europaeus in Northern Italy. AB - We used radiotelemetry to investigate resting sites habitat selection by introduced eastern cottontail (Sylvilagus floridanus) and native European hare (Lepus europaeus) under sympatric conditions. We tracked 24 hares and 34 cottontails in a protected area of northwestern Italy. Hares were found in different sites every week, while cottontails used the same site for two weeks, and occasionally for longer. It is supposed that this periodic nest switching reduces the risk of predation and parasitism. Hares and cottontails forms were located in different habitats and characterized by dense vegetation cover near the ground. This cover increased from winter to summer in both species, while in autumn it continued to increase in cottontails only, and decreased in hares. Cottontails selected shrubby habitats near the river, and avoided crop fields in all seasons. Hares were more adaptive in their search, using high herbs and shrubs all year round, wheat fields in spring, maize in spring and summer, and stubbles in winter. Arguably, partial niche differentiation is necessary to allow the coexistence of similar species. In our study area, hares and cottontails differentiated in the use of resting sites habitats, presumably so as not to compete in this part of their ecological niche. PMID- 21627452 TI - Regulatory mechanism of silkworm hemocyte adhesion to organs. AB - Circulating hemocytes in the body fluid of the silkworm are increased during the larval-larval molting period. We investigated hemocyte adhesion to organs mediating the selectin-selectin ligands during the feeding period and the larval larval molting period using the lectin staining method, sugar chain digestion test with glycoside hydrolases, and the hemocyte adhesion inhibition test using monosaccharides. The results of these tests suggested that the selectin ligand involved in hemocyte adhesion was the Sialyl Lewis x-type, and the structure was changed from the feeding period to the larval-larval molting period. Beta galactosidase appears to be an enzyme that eliminates N-acetylgalactosamine and sialylated N-acetylgalactosamine from the terminal of Sialyl Lewis x. Beta galactosidase activation in skin basement membranes, muscle, fat bodies, midguts, and hemocytes increased markedly during the larval-larval molting period, and at that time, hemocytes were detached from organs. Adding 20-hydroxyecdysone or its analog, tebufenozide to cultured fat bodies increased beta-galactosidase activity in these tissues. Therefore, 20-hydroxyecdysone may induce a structural change in Sialyl Lewis x type sugar chains on the cell surface of silkworm's organs by increasing the beta-galactosidase activity to detach hemocytes from organs and increase the number of circulating hemocytes during the larval-larval molting period. PMID- 21627453 TI - Variations in the arrangement of sensory bristles along butterfly wing margins. AB - The surfaces of insect wings exhibit numerous sensilla, which have been suggested to have a behavioral function. Some evidence suggests that the sensory bristles along the wing margin of lepidopteran insects (butterflies and moths) are involved in the regulation of wing movement. We investigated the arrangement of sensory bristles along the wing margins of 62 species of papilionoid butterflies, using light-microscopic examination of mounts of whole wings after removing the scales surrounding the bristles. In the majority of the wings examined, bristles were located on the ventral wing surfaces and were continuously distributed along the wing margins, except in the vicinity of the wing bases. In some wings, bristles were also located on the dorsal wing surfaces, and were continuously or discontinuously distributed along the wing margins of different species. In a minority of the species studied, we observed bristle distribution in the vicinity of the wing base, discontinuous bristle distribution on both the dorsal and ventral wing surfaces, or an absence of bristles along the wing margins. This variation in the arrangement of bristles along the wing margins is discussed in relation to the reception and transmission of sensory information in the wings. PMID- 21627454 TI - Characterization of an alternative chromatin remodeling to parasperm in a cottid fish, Hemilepidotus gilberti. AB - The dimorphic sperm of Hemilepidotus gilberti, i.e., haploid eusperm and diploid parasperm, have different morphologies corresponding to their own roles in fertilization. To estimate how these specific sperm morphologies were established, we focused on the nuclear morphologies and examined their changing processes in dimorphic spermiogenesis. Electron microscopic observation revealed that, in euspermatids, chromatin condensation first appeared as a mosaic pattern of moderate electrodense material in the peripheral region of the round nucleus. Those materials spread across the whole area to form a uniformly condensed nucleus. Chromatin condensation began similarly in paraspermatids to that in euspermatids. These became localized to one side of a nucleus and further condensed to form strong electrodense chromatin clusters, which are a specific feature of parasperm. From the remodeled nuclei of eusperm and parasperm, we found five and three kinds of sperm-specific basic proteins (SBPs), respectively, substituted to histones. The N-terminus amino acid sequences of the SBPs suggest that, in parasperm, one major SBP and two minor ones were distinct from each other. In eusperm nuclei, two kinds of specific SBPs were detected in addition to the homologs of parasperm SBPs. The specific SBPs had homologous amino acid sequences with huge arginine clusters, and one of them was most dominant among the five kinds of SBPs. The different combinations of SBPs in the eusperm and parasperm may cause a specific pattern of chromatin condensation in the dimorphic sperm nuclei of H. gilberti. PMID- 21627455 TI - Immunolocalization of spetex-1 at the connecting piece in spermatozoa of the musk shrew (Suncus murinus). AB - Spetex-1, which has been isolated by differential display and rat cDNA library screening as a haploid spermatid-specific gene, encodes a protein with two coiled coil motifs that locates at both the segmented column in the connecting piece and outer dense fibers-affiliated satellite fibrils in rat sperm flagella. Orthologs of Spetex-1 are identified in many animal species, including human, chimpanzee, macaque, cow, dog, African clawed frog, green spotted puffer, and zebrafish. In this study, we used RT-PCR in combination with 5' and 3' RACE (Rapid Amplification of cDNA End) technique to isolate Spetex-1 ortholog of the musk shrew (Suneus murinus), which yielded a full-length Suncus Spetex-1 gene containing an open reading frame of 1,908 base pairs encoding a protein of 636 amino acids with the predicted molecular mass of 72,348 Da. Suncus Spetex-1 has two coiled-coil motifs at 118-184 and 242-276 amino acid residues, which is a characteristic shared by mammalian Spetex-1 proteins. To examine the subcellular localization of Spetex-1 in Suncus spermatozoa, we produced the anti-Suncus Spetex-1 antibody and carried out immunocytochemistry. In spite of that the primary structure of Suncus Spetex-1 is basically similar to that of rat and mouse Spetex-1, confocal laser scanning microscopy and immunoelectron microscopy revealed that Spetex-1 was restricted to the segmented column and capitulum in the connecting piece of Suncus spermatozoa and was not detected in other parts of flagella, suggesting a diversity of Spetex-1 localization in mammalian spermatozoa. PMID- 21627456 TI - Unmasking Pachytriton labiatus (Amphibia: Urodela: Salamandridae), with description of a new species of Pachytriton from Guangxi, China. AB - Examination of the lectotype and paralectotypes of Pachytriton labiatus ( Unterstein, 1930 ) from southern China revealed that the specimens do not represent a member of Pachytriton, but are identical with a newt of another genus, Paramesotriton ermizhaoi Wu et al., 2009 also described from southern China. We suggest that Pac. labiatus should be transferred to Paramesotriton as a senior synonym of Par. ermizhaoi. We compared the morphology of the northeastern and southwestern groups of newts previously called Pac. "labiatus" with special reference to age and sexual variations. As a result, we confirmed that the two groups are differentiated sufficiently to be treated as different species. In this report, we revive the name Pac. granulosus Chang, 1933 to refer to the northeastern group of Pac. "labiatus," and at the same time, describe a new species representing the southwestern group. PMID- 21627457 TI - A new didemnid ascidian Lissoclinum midui sp. nov. from Kumejima Island (Okinawa, Japan), with remarks on the absence of a common cloacal system and the presence of an unknown organ. AB - A new photosymbiotic didemnid, Lissoclinum midui sp. nov., is described from coral reefs in the Ryukyu Archipelago, Japan. Colonies of the didemnid are green due to Prochloron algal symbionts, which are distributed solely in the tunic. The new species is placed in Lissoclinum because of its uncoiled vas deferens and the presence of globular spicules. However, two unique characters distinguish this species from all other didemnid ascidians: the absence of a common cloacal system, and the presence of an unknown organ in the bottom wall of the branchial sac. In the phylogenetic trees inferred from partial sequences of cytochrome c oxidase subunit I (COI) gene, the new species diverged at the basal point of the clade of four photosymbiotic Lissoclinum species analyzed here. PMID- 21627458 TI - Effect of non-surgical periodontal therapy on C-reactive protein, oxidative stress, and matrix metalloproteinase (MMP)-9 and MMP-2 levels in patients with type 2 diabetes: a randomized controlled study. AB - BACKGROUND: It is well accepted that glycemic control in patients with diabetes mellitus (DM) is affected by systemic inflammation and oxidative stress. The effect of periodontal therapy on these systemic factors may be related to improvement on glycemic status. The aim of the present study is to assess over a period of 6 months the effect of non-surgical periodontal therapy on serum levels of high-sensitivity C-reactive protein (hsCRP), d-8-iso prostaglandin F2a (d-8 iso) as a marker of oxidative stress, and matrix metalloproteinase (MMP)-2 and MMP-9 on patients with type 2 DM. METHODS: Sixty participants with type 2 DM and moderate to severe periodontal disease were randomized into intervention (IG) and control (CG) groups. IG received scaling and root planing, whereas CG received supragingival cleaning at baseline and scaling and root planing at 6 months. Participants of both groups were evaluated at baseline and 1, 3, and 6 months. Periodontal data recorded at each visit included probing depth, clinical attachment loss, bleeding on probing, and gingival index. Blood was collected at each visit for the assay of serum glycated hemoglobin A1c (A1c), hsCRP, d-8-iso, MMP-2, and MMP-9. RESULTS: Although there was a trend to a reduction in hsCRP, d 8-iso and MMP-9 it did not reach statistical significance. MMP-2 levels remained unchanged after periodontal treatment. CONCLUSION: Effective non-surgical periodontal treatment of participants with type 2 DM and moderate to severe periodontal disease improved significantly A1c levels but did not result in a statistically significant improvement in hsCRP, d-8-iso, MMP-2, and MMP-9 levels. PMID- 21627459 TI - Immediate loading of mandibular overdentures supported by unsplinted direct laser metal-forming implants: results from a 1-year prospective study. AB - BACKGROUND: At present, only some studies have dealt with immediate loading of unsplinted implants supporting mandibular overdentures. The aim of this prospective study is to evaluate treatment outcomes of mandibular overdentures supported by four one-piece, unsplinted, immediately loaded, direct laser metal forming (DLMF) implants by assessing implant survival rate, implant success, marginal bone loss, and prosthetic complications. METHODS: A total of 96 one piece DLMF implants were inserted in the edentulous mandible of 24 patients. Four implants were placed in each edentulous mandible. Immediately after implant placement, a mandibular overdenture was connected to the implants. At 1-year follow-up, clinical, radiographic, and prosthetic parameters were assessed. Success criteria included absence of pain, suppuration, and implant mobility; absence of continuous peri-implant radiolucency; and distance between the implant shoulder and the first visible bone contact <1.5 mm. RESULTS: After a 1-year loading time, the overall implant survival rate was 98.9%, with only one implant lost. Among the surviving 95 implants, two did not fulfill the success criteria; therefore, the implant success rate was 97.8%. The mean distance between the implant shoulder and the first visible bone contact was 0.28 +/- 0.30 mm (95% confidence interval, 0.24 to 0.32). Some prosthetic complications were reported. CONCLUSION: Based on the present results and within the limits of this study, the immediate loading of four unsplinted DLMF implants by means of ball attachment supported mandibular overdentures seems to represent a safe and successful procedure. PMID- 21627460 TI - Are gingival phenotype, residual ridge height, and membrane thickness critical for the perforation of maxillary sinus? AB - BACKGROUND: Sinus membrane perforation is the most common complication of the sinus-augmentation procedure. Anatomic factors have been implicated in the risk of sinus membrane perforation (SMP) during sinus-lift surgery. However, there are no reports on the relationships among gingival phenotype (GP), residual ridge height (RRH), membrane thickness (MT), and perforation rate of the maxillary sinus. Therefore, the aim of this clinical study is to determine the associations among GP, RRH, MT, and SMP rate. METHODS: Forty-four patients (64 sinus-lift procedures) were included in the study. Preoperative computed tomography scanning was performed to radiographically evaluate GP, RRH, and MT. The smallest RRH, highest MT, and gingival thickness values were recorded. These values were classified as follows: RRH, < or >3.5 mm; MT, < or >1 mm; and GP, <1.5 or >2 mm. Sinus-lift surgery was performed with a lateral window approach, and SMPs were detected at 11 sinus sites. Perforation size and locations were recorded, and the perforations were repaired with collagen membrane. One hundred seventy-six implants were placed immediately after augmentation procedure. RESULTS: Strong correlations of GP with RRH (r = 0.722, P = 0.001) and MT (r = 0.702, P = 0.001) were noted. Moderate correlations were found between RRH and MT (r = 0.596, P = 0.001) and between MT and SMP (r = 0.417, P = 0.001). Furthermore, mild correlations of SMP with RRH (r = 0.290, P = 0.020) and GP (r = 0.248, P = 0.04) were established. Four implants were lost, but no significant difference was found between implant failure and SMP. CONCLUSION: Within the limits of the study, it may be suggested that GP, RRH, and MT may be important factors for sinus perforation. PMID- 21627461 TI - Periodontal nitric oxide pathway alteration due to precompetition anxiety in handball players. AB - BACKGROUND: Psychologic and physical stress is known to be related with periodontal disorders. The objective of this study is to investigate salivary and gingival crevicular fluid (GCF) nitric oxide (NO) metabolism and its association with the periodontal reaction to precompetition anxiety in competitive athletes. METHODS: A total of 18 elite male handball players participated in this study. The anxiety level of participants was determined by using a competitive state anxiety inventory-2 (CSAI-2). Periodontal clinical measurements, saliva, and GCF samples were obtained before the application of the CSAI-2 during the half-season break and just before the league championship match. NO-level, arginase, and NO synthase (NOS) activities were analyzed in saliva samples; NO and basic fibroblast growth factor (FGF-b) levels were analyzed in GCF samples. RESULTS: The CSAI-2 was significantly different between the two evaluation time points. GCF NO and FGF-b levels increased before the championship match, whereas saliva levels did not change. The plaque index and gingival index (GI) increased at the evaluation just before the championship match. The difference in the GI was statistically significant. CONCLUSION: The competitive stress of athletes may lead to a gingival/periodontal disturbance accompanied by an altered NO metabolism in saliva and GCF. PMID- 21627462 TI - A retrospective analysis of implants immediately placed in sites with and without periapical pathology in sixty-four patients. AB - BACKGROUND: Many patients requiring implant therapy present with hopeless teeth exhibiting periapical pathology. The advisability of implant placement in such situations has not been conclusively determined. METHODS: Sixty-four patients underwent therapy in their maxillary incisor region. Treatment consisted of immediate implant placement in a site demonstrating periapical pathology, and immediate implant placement in a "pristine" site, either during the same visit or during separate visits. The implants placed in the sites demonstrating periapical pathology were followed in function for <=117 months, with a mean time in function of 64 months. The implants placed in pristine sites were followed in function for <=120 months, with a mean time in function of 62 months. RESULTS: Two implants in the central incisor positions of one patient demonstrated 2 mm of buccal recession after ~46 months in function. These implants were deemed esthetic failures, despite the absence of inflammation and continued clinical implant immobility, yielding cumulative survival rates of 98.1 and 98.2 for implants placed in sites with periapical pathology and implants placed in sites without periapical pathology, respectively, according to published criteria. CONCLUSIONS: Implants immediately placed in sites demonstrating periapical pathology yielded results comparable to those immediately placed in pristine sites. The difference in survival rates was not statistically significant. PMID- 21627463 TI - The effect of smoking on periodontal bone regeneration: a systematic review and meta-analysis. AB - BACKGROUND: The effect of smoking on soft tissues after periodontal treatment has been extensively studied. However, little focus has been placed on the impact of smoking on bone regeneration after treatment. The aim of this review is to systematically assess the effect of smoking on bone regeneration after periodontal treatment. METHODS: A protocol was established and studies were sourced from five electronic databases. Screening, data abstraction, and quality assessment was conducted by two review authors. Prospective and retrospective clinical studies assessing bone regeneration in smokers and non-smokers after periodontal therapy were selected. In addition, arms of clinical trials comparing different interventions that reported results separately for smokers and non smokers were also included. Primary outcome measures were based on clinical and/or radiographic indicators of bone regeneration after periodontal therapy. The review and meta-analysis followed many of the recommendations outlined in the preferred reporting items for systematic reviews and meta-analyses statement. RESULTS: Six of 10 studies included in this review concluded that smoking negatively influenced bone regeneration. A meta-analysis of a subgroup of three studies demonstrated that smoking resulted in significantly less bone gain (P = 0.03) as measured by a change in the probing bone level after the treatment of intrabony defects with guided tissue regeneration. The meta-analysis showed a standardized mean difference of -2.05 (95% confidence interval: -2.64 to -1.47) using the random-effects model. CONCLUSIONS: Smoking has a negative effect on bone regeneration after periodontal treatment. Patients should be advised that their smoking habit may result in poorer bone regeneration after periodontal treatment. PMID- 21627464 TI - Transcript profiling and gene characterization of three fatty acid desaturase genes in high, moderate, and low linolenic acid genotypes of flax (Linum usitatissimum L.) and their role in linolenic acid accumulation. AB - Three genes encoding fatty acid desaturase 3 (fad3a, fad3b, and a novel fad3c) were cloned from four flax genotypes varying in linolenic acid content. Real-time PCR was used to quantify expression levels of the three fad3 genes during seed development. High amounts of both fad3a and fad3b transcripts were observed and reached their peak levels at 20 days after anthesis, except for fad3a from SP2047 where only low level expression was observed throughout seed development. Transcript accumulation of the novel fad3c gene was at similar background levels. The fatty acid composition was analysed for all genotypes and stages of development and compared with the fad3 gene expression patterns. alpha-Linolenic acid gradually accumulated during seed development, while linoleic acid was transient and decreased in M5791, UGG5-5, and AC McDuff. In contrast, the linolenic acid present in the early stages of development nearly completely disappeared in SP2047, while linoleic acid steadily accumulated. fad3a of the low linolenic acid line SP2047 encoded a truncated protein caused by a premature stop codon resulting from a single point mutation, and the low level of transcript accumulation in this genotype is likely due to nonsense-mediated mRNA decay caused by the premature termination of translation as a result of this early stop codon. Although substantial amounts of transcript accumulation occurred with fad3b of SP2047 genotype, cloning of the gene revealed a mutation in the first histidine box causing an amino acid change. Heterologous expression in yeast of the SP2047 and UGG5-5 fad3b genes showed that the mutation in the histidine box in SP2047 caused the enzyme inactivity. Taken together, these results showed that fad3a and fad3b are responsible for linolenic acid accumulation in flax seeds but did not support a major role for the novel fad3c. These observations were further supported by phenotypic and genotypic assessment of a doubled haploid population. Expression patterns of fad3a and fad3b were highly correlated with linolenic acid accumulation during seed development, with the exception of fad3b in SP2047 whose lack of activity was caused by the histidine box mutation despite its transcript accumulation being similar to that of the fad3b of the other genotypes. PMID- 21627465 TI - Transcriptional regulation of ompF2, an ompF paralogue, in Yersinia pestis. AB - A regulatory circuit composed of three porins (OmpF, OmpC, and OmpX) and two transcriptional regulators (OmpR and CRP) has previously been characterized in Yersinia pestis . In this follow-up study, OmpF2, an OmpF paralogue, was integrated into this regulatory circuit. Only basal expression was detected for ompF2 in the wild-type strain under different osmotic conditions. The ompF2 transcription was dramatically enhanced with increasing medium osmolarity in the ompR null mutant background. The CRP regulator had no regulatory effect on ompF2 under the growth conditions tested. PMID- 21627466 TI - Aptamer selection for the detection of Escherichia coli K88. AB - In this study, the first group of single-stranded DNA aptamers that are highly specific to enterotoxigenic Escherichia coli (ETEC) K88 was obtained from an enriched oligonucleotide pool by the SELEX (Systematic Evolution of Ligands by Exponential Enrichment) procedure, during which the K88 fimbriae protein was used as the target and bovine serum albumin as counter targets. These aptamers were applied successfully in the detection of ETEC K88. They were then grouped under different families based on the similarity of their secondary structure and the homology of their primary sequence. Four sequences from different families were deliberately chosen for further characterization by fluorescence analysis. Having the advantage of high sensitivity, fluorescence photometry was selected as single stranded DNA quantification method during the SELEX process. Aptamers with the highest specificity and affinity were analyzed to evaluate binding ability with E. coli. Since ETEC K88 is the only type of bacterium that expressed abundant K88 fimbriae, the selected aptamers against the K88 fimbriae protein were able to specifically identify ETEC K88 among other bacteria. This method of detecting ETEC K88 by aptamers can also be applied to bacteria other than ETEC K88. PMID- 21627467 TI - Hybrid techniques and neuraxial modulation for treatment of ventricular tachycardia. PMID- 21627468 TI - To use or not to use: a focus on endoscopic vein harvesting. PMID- 21627469 TI - Future anticoagulants in interventional cardiology: anti-IXa and anti-Xa agents in percutaneous coronary intervention. PMID- 21627471 TI - Quest for lower immunosuppression in cardiac transplantation: an analysis of the TICTAC trial. AB - Evaluation of: Baran DA, Zucker MJ, Arroyo LH et al.: A prospective, randomized trial of single versus dual drug immunosuppression in heart transplantation: the Tacrolimus in Combination, Tacrolimus Alone Compared (TICTAC) trial. Circ. Heart Fail. 4(2), 129-137 (2011). Success in cardiac transplantation has been attributed to improved immunosuppressive regimens. Conventionally, patients are treated with a triple regimen following transplantation, consisting of a calcineurin inhibitor (cyclosporine or tacrolimus), antiproliferative agent (mycophenolate mofetil) and corticosteroids. These agents, however, are associated with a significant morbidity which has prompted a quest for minimization of immunosuppressive regimens; a challenging goal, given the potential for allograft rejection particularly early after transplantation. This article reviews a recent study by Baran and colleagues who boldly challenged the notion that triple immunosuppression is a prerequisite for success after cardiac transplantation. The authors successfully demonstrate the feasibility of weaning corticosteroids and mycophenolate mofetil early after cardiac transplantation, maintaining patients solely on monotherapy with tacrolimus. This evaluation focuses on the findings and limitations of the study and provides the historical background which led to this promising clinical trial. PMID- 21627472 TI - Advances in cardiology: clinical trial update. AB - Multiple key cardiology trials have been presented or published over recent months, several with the potential to change clinical practice. In this article, we summarize and place in clinical context new trial findings regarding anticoagulation in the cardiac catheterization laboratory (enoxaparin, fondaparinux and unfractionated heparin), the implications of genetic polymorphisms and functional testing for antiplatelet therapy (clopidogrel and ticagrelor), new oral anticoagulants for use in atrial fibrillation (apixiban and rivaroxaban), optimal pacing strategies and pharmacological agents in heart failure (ivabradine, eplerenone, cardiac resynchronization therapy, telemonitoring and intracoronary bone marrow stem cell infusion). Clinical trials in percutaneous structural intervention (transcatheter aortic valve implantation, MONARCTM mitral annular implant, STARFlex((r)) patent foramen ovale device) and advanced percutaneous coronary intervention (everolimus-eluting stents, biodegradable polymer/polymer-free technologies and contemporary use of intravascular ultrasound) are also discussed. PMID- 21627473 TI - Awareness of heart attack symptoms among US adults in 2007, and changes in awareness from 2001 to 2007. AB - OBJECTIVE: Timely access to emergency care, prompt receipt of advanced treatment and survival from heart attack is dependent on both the early recognition of heart attack symptoms, by both victims and bystanders, and by immediately calling the emergency services. The objective of this study is to measure the awareness of heart attack symptoms and the emergency response among US adults. METHOD: We analyzed data from the Behavioral Risk Factor Surveillance System's module on heart attack and stroke, which was conducted in 17 states/territories in 2001 and 12 states/territories in 2007. The module included five questions related to heart attack symptoms, one decoy question and one question regarding the first action to take if someone is having a heart attack. Age-adjusted prevalence of awareness was estimated, and odds ratios were calculated. Differences between 2001 and 2007 were assessed for five states that used the module in both years. RESULTS: In 2007, among 76,864 adults, awareness of individual heart attack symptoms ranged from 49% (pain in jaw, neck or back) to 92% (chest pain). Although 97% of adults recognized at least one symptom, only 10.7% recognized all five symptoms, knew that 'sudden trouble seeing in one or both eyes' was not a symptom of heart attack and recognized the need to call the emergency services. This estimate varied significantly by age, sex, race/ethnicity and level of education. The estimate was significantly higher for women (12.2%) than men (7.7%), White (11.6%) than Black (5.7%) or Hispanic people (4.5%), those with a higher level of education (13.5%) than lower educational level (4.5%) and for those with coronary heart disease (16.2%) than without the disease (9.5%). Comparison of awareness between 2001 (the referent) and 2007, in five states, revealed that awareness of all symptoms and calling the emergency services, were 9.7 and 10.3% for 2001 and 2007, respectively (p < 0.01). Compared to 2001, the odds ratio of awareness in 2007 was 1.08 (95% CI: 0.99-1.19) after controlling for socioeconomic and clinical characteristics. CONCLUSION: Awareness of all heart attack symptoms among adults in 12 states was low and little improvement was observed between 2001 and 2007. Accordingly, both clinicians and public health officials should seek ways in order to increase public awareness of the symptoms of heart attack. Special efforts should be focused on men, Black and Hispanic people and those with only modest levels of education. PMID- 21627474 TI - Incidence, prognostic value and management of vascular complications with transfemoral transcatheter aortic valve implantation. AB - Inoperable or high-risk patients with severe aortic stenosis who undergo transcatheter aortic valve implantation (TAVI) have better outcomes compared with those treated with standard medical therapy. As for any other invasive procedure, peri-procedural complications may occur, reducing the procedural success rate and potentially affecting short- and mid-term outcomes. The transfemoral approach prevails over other possible access-site options in most registries. The use of large introducer sheaths and the need for double arterial vascular access can lead to higher rates of vascular complications in this elderly population, with a high prevalence of baseline peripheral artery disease. In this article, we review the results of recent clinical trials and major registries using the two different bioprosthesis currently available for TAVI, focusing on access site related complications with transfemoral TAVI, their management and relationship with in-hospital and 30-day survival. Awareness of the mechanisms behind these complications might help in their prevention, recognition and management and may ultimately improve the clinical outcome of TAVI procedures. PMID- 21627476 TI - Assessment of cardiac dynamics during stress echocardiography by the peak power output-to-left ventricular mass ratio. AB - Peak cardiac power-to-mass and peak mass-to-power are variables that couple cardiac power output with left ventricular (LV) mass at peak exercise or during maximal inotropic stimulation. Quantitative stress echocardiography enables the calculation of power output according to the formula: 133 * 10(-6) * stroke volume per second (ml) * mean blood pressure (BP; mmHg) * heart rate. Power-to mass may be calculated as LV power output per 100 g of LV mass: 100 g * LV power output divided by LV mass (W/100 g). Conversely, mass-to-power may be estimated by dividing LV mass index by LV power output (g/m(2)/W). With a little rearrangement of the formulas we can write: power-to-mass (W/100 g) = 0.222 * cardiac output (l/min) * mean BP (mmHg)/LV mass (g) and mass-to-power (g/m(2)/W) = LV mass index/0.00222 * cardiac output (l/min) * mean BP (mmHg). These parameters reflect the energy delivery of ventricular myocardium with respect to potential energy that is stored in LV mass. The assessment of peak power-to-mass and peak mass-to-power indices may be useful to distinguish compensatory versus maladaptive remodeling in patients with LV dysfunction. When the integrity of myocardial structure is compromised, a disproportion becomes apparent between maximal cardiac power output and LV mass and this leads to either a reduction of peak power-to-mass or an increase of peak mass-to-power. Preliminary reports have demonstrated the usefulness and the prognostic value of peak power-to-mass and peak mass-to-power in patients with LV systolic dysfunction and coronary artery disease. PMID- 21627475 TI - The golden age of minimally invasive cardiothoracic surgery: current and future perspectives. AB - Over the past decade, minimally invasive cardiothoracic surgery (MICS) has grown in popularity. This growth has been driven, in part, by a desire to translate many of the observed benefits of minimal access surgery, such as decreased pain and reduced surgical trauma, to the cardiac surgical arena. Initial enthusiasm for MICS was tempered by concerns over reduced surgical exposure in highly complex operations and the potential for prolonged operative times and patient safety. With innovations in perfusion techniques, refinement of transthoracic echocardiography and the development of specialized surgical instruments and robotic technology, cardiac surgery was provided with the necessary tools to progress to less invasive approaches. However, much of the early literature on MICS focused on technical reports or small case series. The safety and feasibility of MICS have been demonstrated, yet questions remain regarding the relative efficacy of MICS over traditional sternotomy approaches. Recently, there has been a growth in the body of published literature on MICS long-term outcomes, with most reports suggesting that major cardiac operations that have traditionally been performed through a median sternotomy can be performed through a variety of minimally invasive approaches with equivalent safety and durability. In this article, we examine the technological advancements that have made MICS possible and provide an update on the major areas of cardiac surgery where MICS has demonstrated the most growth, with consideration of current and future directions. PMID- 21627477 TI - Future of personalized pharmacotherapy in chronic heart failure patients. AB - There is a significant amount of diversity among heart failure (HF) patients. Contemporary HF regimens often do not take into consideration many of the factors that might influence an individual's response to treatment. Clinical recommendations based on trial data derived from mainly younger Caucasian male study populations have, in most cases, been applied equally to women and African Americans. Subgroup analyses of randomized HF trials and results of retrospective cohort studies have been used for customizing HF regimens in women and African Americans. Pharmacogenetics is an emerging strategy for personalizing HF therapy. Genetic biomarkers may play an important role in predicting a patient's response to treatment and in predicting those at risk of toxicity. HF pharmacotherapy has improved over the last two decades; however, substantial work remains in order to personalize HF management and maximize the benefit of pharmacologic interventions, while limiting adverse events. PMID- 21627478 TI - Antiplatelet drug therapy: role of pharmacodynamic and genetic testing. AB - Antiplatelet therapy represents the cornerstone of treatment for the short- and long-term prevention of atherothrombotic disease processes, in particular in high risk settings such as in patients with acute coronary syndrome and those undergoing percutaneous coronary intervention. Currently, dual antiplatelet therapy with aspirin and clopidogrel represents the most commonly used treatment regimen in these settings. However, a considerable number of patients continue to experience adverse outcomes, including both bleeding and recurrent ischemic events. Numerous investigations have demonstrated that this phenomenon may be, in part, attributed to the broad variability in individual response profiles to this standard antiplatelet treatment regimen, as identified by various assays of platelet function testing. In addition, recent studies have demonstrated that genetic polymorphisms may also have an important role in determining levels of platelet inhibition and may be considered as a tool to identify patients at risk of adverse events. This article provides an overview on antiplatelet drug response variability, an update on definitions, including the role of pharmacodynamic testing, underlying mechanisms - with emphasis on recent understandings on pharmacogenetics and drug-drug interactions - and current and future perspectives on individualized antiplatelet therapy. PMID- 21627479 TI - Recent developments in the use of antiplatelet agents to prevent cardiovascular events. AB - Platelets are pivotal contributors to arterial thrombosis. Dual antiplatelet therapy with aspirin and clopidogrel has become the standard of care for the secondary prevention of cardiovascular events in patients with acute coronary syndromes and after percutaneous coronary intervention. Clinical evidence of the continued risk of cardiovascular events plus pharmacodynamic evidence of substantial variability in on-treatment platelet reactivity has supported the development of new therapeutic strategies, using established agents and new antiplatelet drugs. This article will highlight recent pivotal clinical trials seeking to advance the use of antiplatelet therapy in patients with cardiovascular disease. PMID- 21627480 TI - Atrial fibrillation in the aging heart: pharmacological therapy and catheter ablation in the elderly. AB - The majority of patients with atrial fibrillation (AF) seeking medical treatment are in the elderly age group and the management of these patients is often complicated by comorbidities, challenging the pharmacological management of these patients. Owing to hypertension, congestive heart failure, left ventricular hypertrophy and coronary artery disease, antiarrhythmic treatment often fails due to side effects, proarrhythmia or poor rhythm control. In recent years, radiofrequency catheter ablation has been widely performed as an effective treatment for recurrent, drug-refractory AF. However, few elderly patients were included in prior AF catheter ablation studies and the current guidelines for catheter ablation of AF recommend a conservative approach in the elderly population owing to the absence of clinical data. However, study results from our group and others suggest that catheter ablation is a safe and effective treatment for patients over the age of 65 years with symptomatic, drug-refractory AF and, therefore, patients should not be excluded from catheter ablation on the basis of age alone. In this article, we discuss the pharmacological (rhythm control, rate control and anticoagulation) and catheter management of AF in the elderly population. PMID- 21627481 TI - Mental stress and ischemic heart disease: evolving awareness of a complex association. AB - The connection between cardiovascular disease and psychosocial risk factors has been the subject of an ever-growing body of literature over the last 50 years. Studies on the role of negative emotions, personality traits, chronic stress and social determinants have brought to light their possible role in triggering acute coronary syndromes, although further studies are required to clarify controversial results regarding the association between cardiovascular risk and important psychological problems such as depression and anxiety. The recognition of the role of emotional events in acute coronary syndromes paved the way for provocation experiments, aimed at inducing mental stress in a controlled setting and then documenting reversible impairment of myocardial perfusion, depolarization anomalies and arrhythmias. This ultimately led to the formalization of the concept of mental stress-induced myocardial ischemia. Accumulating evidence on the mechanistic bases of such phenomena outline a wide range of central and peripheral physiological changes associated with emotions and behaviors, whose effects are exerted on the cardiovascular system, sympathetic nervous system and the hypothalamus-hypophysis neuroendocrine axis. This article outlines the main steps in the identification of psychological aspects as cardiovascular risk factors and emphasizes the relevance of emotional stress as a trigger of acute cardiovascular events. Finally, a description is provided of the pathophysiological mechanisms behind mental stress-induced myocardial ischemia and pathways connecting the heart and brain. PMID- 21627485 TI - Simulation of differential drug pharmacokinetics under heat and exercise stress using a physiologically based pharmacokinetic modeling approach. AB - Under extreme conditions of heat exposure and exercise stress, the human body undergoes major physiological changes. Perturbations in organ blood flows, gastrointestinal properties, and vascular physiology may impact the body's ability to absorb, distribute, and eliminate drugs. Clinical studies on the effect of these stressors on drug pharmacokinetics demonstrate that the likelihood of pharmacokinetic alteration is dependent on drug properties and the intensity of the stressor. The objectives of this study were to use literature data to quantify the correlation between exercise and heat exposure intensity to changing physiological parameters and further, to use this information for the parameterization of a whole-body, physiologically based pharmacokinetic model for the purposes of determining those drug properties most likely to demonstrate altered drug pharmacokinetics under stress. Cardiac output and most organ blood flows were correlated with heart rate using regression analysis. Other altered parameters included hematocrit and intravascular albumin concentration. Pharmacokinetic simulations of intravenous and oral administration of hypothetical drugs with either a low or high value of lipophilicity, unbound fraction in plasma, and unbound intrinsic hepatic clearance demonstrated that the area under the curve of those drugs with a high unbound intrinsic clearance was most affected (up to a 130% increase) following intravenous administration, whereas following oral administration, pharmacokinetic changes were smaller (<40% increase in area under the curve) for all hypothetical compounds. A midazolam physiologically based pharmacokinetic model was also used to demonstrate that simulated changes in pharmacokinetic parameters under exercise and heat stress were generally consistent with those reported in the literature. PMID- 21627486 TI - Class 1 and class 2 integrons in multidrug-resistant gram-negative bacteria isolated from the Salmon River, British Columbia. AB - Using an enrichment protocol, we isolated 16 gram-negative, multidrug-resistant strains of known or opportunistic bacterial pathogens from the Salmon River in south-central British Columbia from 2005 to 2009, and investigated the genetic basis of their resistance to a variety of antibiotics. Of the 16 strains, 13 carried class 1 integrons and three carried class 2 integrons. Genes found in cassettes associated with the integrons included those for dihydrofolate reductases (dfrA1, dfrA12, dfrA17, and dfrB7), aminoglycoside adenyltransferases (aadA1, aadA2, aadA5, and aadB), streptothricin acetyltransferase (sat), and hypothetical proteins (orfF and orfC). A new gene cassette of unknown function, orf1, was discovered between dfrA1 and aadA5 in Escherichia sp. Other genes for resistance to tetracycline, chloramphenicol, streptomycin, and kanamycin (tetA, tetB, tetD; catA; strA-strB; and aphA1-Iab, respectively) were outside the integrons. Several of these resistance determinants were transferable by conjugation. The detection of organisms and resistance determinants normally associated with clinical settings attest to their widespread dispersal and suggest that regular monitoring of their presence in aquatic habitats should become a part of the overall effort to understand the epidemiology of antibiotic resistance genes in bacteria. PMID- 21627487 TI - Epidemiological and molecular characterization of Staphylococcus haemolyticus strains, from a hematology ward, with decreased susceptibility to glycopeptides. AB - In the present study, we report on the reduced susceptibility to teicoplanin among clinical isolates of Staphylococcus haemolyticus in a hematology ward of a teaching hospital. The molecular characterization of 17 S. haemolyticus strains was performed using mec gene complex classification, pulsed-field gel electrophoresis analysis, and minimum inhibitory concentration examination. Pulsotype A strains carrying a class C2 mec gene complex were the most prevalent strains, at 64.7%. In vivo selection of stepwise increase in resistance to vancomycin and teicoplanin was observed in three S. haemolyticus strains serially isolated from a case patient. The results of the present study suggest the regional spread of certain S. haemolyticus clones with diminished susceptibility to glycopeptides, emphasizing the need for continuous monitoring of minimum inhibitory concentration levels of vancomycin and teicoplanin in S. haemolyticus strains, and the importance of infection control practices to prevent its transmission. PMID- 21627488 TI - A pilot assessment of oxidative stress byproducts and antioxidant activities among Indian patients with various stages of hypertension. AB - In humans, hypertension is considered a state of oxidative stress that can contribute to the development of atherosclerosis and other hypertension-induced organ damages. The objective of this study was to evaluate oxidative status, antioxidant activities, and oxidative stress by-products among Indian patients with various stages of hypertension. Lipid profile, enzymatic and non-enzymatic antioxidants, lipid peroxidation as thiobarbituric acid reactive substances (TBARS), C-reactive protein, electrolytes, and minerals were analyzed in the blood of newly diagnosed prehypertensives, stage I and II hypertensives (n = 20 in each group) and were compared to their age-matched normotensives. Elevated levels of lipid profile (except high density lipoprotein cholesterol [HDL-C]) were observed in stage I and II hypertensive patients. Enzymatic and non enzymatic antioxidants were significantly (P < 0.05) lower, while TBARS and C reactive protein were higher in prehypertensives, and stage I and II hypertensives. Significant (P <0.05) changes were also observed in the plasma Na(+) and K(+) concentrations among the hypertensive groups. Serum levels of zinc, copper, and magnesium were significantly (P < 0.05) lower in prehypertensives, and stage I and II hypertensives as compared to normotensives. The study indicated a strong association between blood pressure (BP) and oxidative stress-related parameters and suggests a possible role of oxidative stress in the development of elevated BP. PMID- 21627489 TI - A 29-gauge atraumatic needle for amniocentesis. AB - OBJECTIVE: To compare perforation characteristics of standard 22 G (0.7 mm) to 29 G needle (0.34 mm) for amniocentesis. METHODS: Seventeen human chorio-amnion membranes were perforated immediately after cesarean section using 22 G needle for spinal anesthesia and 29 G "pencil-point" needles for amniocentesis under in vitro conditions. Area of perforation was determined using a microscope and volume of fluid leakage was measured over a period of 5 min. RESULTS: Membrane perforation with the 22 G needle resulted in a mean damaged area of 225,147.4 MUm(2), a hole with a mean area of 50,154 MUm(2) and amniotic fluid volume passage of 17.5 mL/5 min, whereas the 29 G needle generated a mean damaged area of 114,812.4 MUm(2), a hole with an average area of 1382.5 MUm(2) and volume passage of 0.28 mL/5 min. These differences were significant. CONCLUSION: The hole formed by membrane perforation with 29 G "pencil-point" needle for amniocentesis is 36 times smaller, and the amniotic fluid loss is 61 times less than that measured with the 22 G standard needle for spinal anesthesia. Significant reduction of complications following amniocentesis is expected with the 29 G needle. PMID- 21627490 TI - Candidates for the determination of antithrombin activity in pregnant women. AB - BACKGROUND: Some women exhibit a gradual decrease in antithrombin activity until the time of delivery, with antithrombin activity reaching <65% of the normal level (pregnancy-induced antithrombin deficiency, PIATD). However, the clinical features of such women are not well understood and are unfamiliar to many obstetricians. FINDINGS: PIATD is more likely to occur as the number of fetuses increases (approx. 1.0%, 10%, and 40% for singleton, twin, and triplet pregnancies, respectively) and in women with hypertension, isolated proteinuria and/or edema. However, normotensive women account for 60% of women with PIATD. Antithrombin can escape from the blood into the interstitial space. Women with PIATD suffer from a decreased plasma volume and are more likely to develop liver dysfunction, irrespective of the presence or absence of hypertension. Because antithrombin activity continues to decrease until the time of delivery in women with PIATD, women with unrecognized PIATD may be identified as patients with so called "acute fatty liver of pregnancy" if delivery is delayed. CONCLUSION: Knowledge of AT activity in obstetrical practice may improve management of pregnant women. The determination of AT activity should be considered in women with multifetal pregnancies, hypertension, isolated proteinuria and/or edema. PMID- 21627491 TI - Vaginal breech delivery in very low birth weight (VLBW) neonates: experience of a single center. AB - AIMS: To compare the short-term maternal and neonatal outcomes of very low birth weight (VLBW) breech singletons by mode of delivery. METHODS: All breech fetuses born from 24-0/7 to 26-6/7 weeks' gestation at our institution between 2000 and 2008 were eligible for the study. Abstracted medical record data included maternal demographics, delivery data, and neonatal outcomes. RESULTS: There were 26 vaginal and 39 cesarean deliveries. Maternal age did not differ between groups; gestational age was greater in the cesarean group by five days. Short term neonatal outcomes did not differ between groups. Of the 39 cesarean deliveries, 27 involved classical uterine incisions. Estimated blood loss (732 mL vs. 362 mL) and postpartum infection rate (26% vs. 4%) were greater with cesarean delivery. CONCLUSION: Neonatal outcome is not improved in VLBW infants born by cesarean section. Given the morbidity of classical cesarean sections, vaginal delivery of the breech VLBW infant may be safely considered. PMID- 21627492 TI - Calcium pyrophosphate and monosodium urate crystals in synovial fluid as a cause of pseudoeosinophilia. AB - BACKGROUND: Synovial fluid (SF) leukocytes can be counted microscopically in a Neubauer chamber or by automated procedures using haematology analysers. Knowledge of laboratory artefacts is crucial for the correct interpretation of results obtained using automated methods. SF pseudoeosinophilia has recently been described as a new artefact in patients with crystal-related arthropathies. We investigated whether pseudoeosinophilia of SF is restricted to crystal-related disorders, or if it may also occur in other arthropathies. METHODS: We compared the percentages of eosinophils in 120 crystal containing SF samples with 185 crystal-free specimens using the Wilcoxon test. Crystal positive samples, determined by polarised microscopy, contained at least two monosodium urate or calcium pyrophosphate crystals per 10 high power fields (630* magnification). True SF eosinophilia was ruled out by microscopic examination of stained slides. RESULTS: Crystal positive samples had significantly higher percentages of eosinophils than the controls (p<0.0001). No significant differences between the two crystal types were found (p=0.693). Thus, pseudoeosinophilia was significantly correlated with the presence of crystals, and none of the distinct crystal types was more likely to be associated with pseudoeosinophilia. CONCLUSIONS: In this study, SF pseudoeosinophilia was confirmed as a crystal related laboratory artefact which has to be considered in the interpretation of automated SF leukocyte differential counts. PMID- 21627493 TI - Evaluation of a commercially available rapid urinary porphobilinogen test. AB - BACKGROUND: Demonstration of substantially increased urinary excretion of porphobilinogen is the cornerstone of diagnosing acute porphyria crisis. Because porphobilinogen testing is not implemented on clinical chemistry analysers, respective analyses are available in rather few clinical laboratories. The aim of this study was to critically describe and to evaluate a semi-quantitative rapid test for urinary porphobilinogen determination which is commercially available and recommended by the American Porphyria Foundation. METHODS: Urinary samples from patients with acute intermittent porphyria and control samples were analysed and the semi-quantitative results were compared with the results obtained by a manual quantitative spectrophotometric method. RESULTS: In all 32 samples studied, acceptable agreement between the results of the rapid test and the quantitative test was observed. Handling of the test was found to be convenient. CONCLUSIONS: The assay was found to be reliable and has the potential to increase the availability of porphobilinogen testing in the field. PMID- 21627494 TI - Laboratory assessment of iron status in pregnancy. AB - BACKGROUND: Efforts to improve maternal nutrition during pregnancy prompted an observational study of the occurrence of maternal iron deficiency and its laboratory diagnosis in almost 500 pregnancies. METHODS: In this longitudinal study, the biochemical and haematological iron indices of women (n=492) attending a prenatal clinic in a Dublin maternity hospital were assessed at first booking (mean 15.9 weeks), and after 24 weeks, and 36 weeks of gestation. Full blood counts were measured. Serum ferritin (SF), zinc protoporphyrin (ZPP), and transferrin receptor (sTfR) concentrations were assayed and transferrin receptor index (sTfR-Index) was calculated. The occurrence of low values and their diagnostic values were considered. RESULTS: A high occurrence iron deficiency (ID) at first booking (SF<12 MUg/L) had increased over six-fold by 24 weeks, and all biochemical iron indices reflected progressive iron depletion right up to term. The WHO recommended anaemia "cut-off" (Hb<110 g/L) was insensitive to biochemical iron deficiency at booking, missing over 90% of the low SF values (SF<12 MUg/L) which were mostly associated with much higher Hb levels. CONCLUSIONS: This study stresses the importance of including a biochemical index of iron status in prenatal screening and supports SF as the best indicator of biochemical ID overall. sTfR was insensitive to iron deficiency in early pregnancy, whereas the sTfR-Index, as a ratio, has the potential to distinguish between ID and physiological anaemia, and may offer stability in the assessment of iron stores from early pregnancy to full term. A policy of early screening of both Hb and SF concentrations is recommended as the minimum requirement for surveillance of maternal iron status in pregnancy. PMID- 21627495 TI - A template for mutational data analysis of the CFTR gene. AB - BACKGROUND: Automated DNA sequencing produces large amounts of data that need to be analyzed by appropriate software. Personalization of software can be a difficult and time-consuming task, especially if a large number of mutations have to be analyzed. METHODS: The Applied BioSystems SeqScape software, based on the KB basecaller algorithm, is a versatile tool that can be used for mutational analysis and for data quality assessment of sequences belonging to any gene of interest. Using this software we analyzed over 1400 sequences of CFTR exons and adjacent intronic zones, representing over 500,000 bases. RESULTS: We present an up to date specific template and a linked set of instructions for automated labeling of all point mutations and polymorphisms of the CFTR gene, whose mutations cause cystic fibrosis (the most common genetic disease among Caucasian individuals). We also describe our refined software settings for mutational analysis, in order to keep to a minimum the need of manual validation. CONCLUSIONS: The use of our template greatly simplifies the mutational analysis of the CFTR gene, reducing human intervention. In our opinion, it might not only be useful to researchers that already perform CFTR mutational analysis by sequencing methods but it should also improve the approach in those laboratories that already use ABI PRISM instrumentation for a limited mutational analysis of the CFTR gene. Similar mutational templates can also be used for other disease causing genes, thus improving molecular genetics protocols. PMID- 21627498 TI - Questions depth of anesthesia and adequacy of ventilation in fish article. Letters to the editor. PMID- 21627499 TI - What is your diagnosis? Cardiac hemangiosarcoma. PMID- 21627500 TI - What is the evidence? Combination of local analgesic and opioid epidural protocals. PMID- 21627501 TI - Pathology in practice. Hypertrophic osteodystrophy. PMID- 21627502 TI - Pathology in practice. Severe, chronic, segmental proliferative and ulcerative enteritis with intraepithelial curved bacilli (L intracellularis) and multifocal transmural necrosis. PMID- 21627503 TI - Comparing means or distributions. PMID- 21627504 TI - Update on Salmonella spp contamination of pet food, treats, and nutritional products and safe feeding recommendations. PMID- 21627505 TI - Impact of gender and race-ethnicity on reasons for pursuing a career in veterinary medicine and career aspirations. AB - OBJECTIVE: To determine the impact of gender and race-ethnicity on reasons applicants to the Purdue University School of Veterinary Medicine decided to pursue a career in veterinary medicine and their career aspirations. DESIGN: Retrospective cross-sectional study. SAMPLE: Personal statements from 694 veterinary medical school applications submitted in 2008. PROCEDURES: Personal statements were read by investigators to identify the turning point for each applicant's decision to pursue a career in veterinary medicine and each applicant's intended career path. RESULTS: Veterinary practice experience and animal ownership were the most frequently stated reasons for pursuing a veterinary career; differences were not identified between males and females. More Caucasian applicants than underrepresented minority (URM) applicants stated veterinary practice experience and more URM applicants than Caucasian applicants cited animal ownership as a reason for pursuing a veterinary career. Many applicants did not cite a specific career path within veterinary medicine; applicants who indicated a career path most often cited veterinary practice. More females than males stated an interest in equine medicine, and more Caucasian applicants than URM applicants indicated an interest in mixed animal practice. More URM applicants than Caucasian applicants indicated a desire to pursue specialty training. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that veterinary practice experience and animal ownership were important factors influencing applicants' decision to pursue a veterinary career, but many applicants had not selected a specific career path. Opportunities exist to influence the decisions of individuals to become veterinarians and the selection of specific career paths within the veterinary profession. PMID- 21627507 TI - Evaluation of twice-daily lower-dose trilostane treatment administered orally in dogs with naturally occurring hyperadrenocorticism. AB - OBJECTIVE: To evaluate effectiveness and incidence of adverse reactions to twice daily lower-dose oral administration of trilostane in the treatment of dogs with naturally occurring hyperadrenocorticism (NOH). DESIGN: Clinical trial. ANIMALS: 47 dogs with NOH. PROCEDURES: 47 dogs were treated orally with trilostane (0.21 to 1.1 mg/kg [0.1 to 0.5 mg/lb], q 12 h). All dogs were reevaluated at 2 weeks and 2 months, 38 dogs at 6 months, and 28 dogs at 1 year of treatment. RESULTS: 9 of 47 dogs had an adrenocortical tumor causing NOH, and all had good responses after 2 months (mean trilostane dosage, 0.89 mg/kg [0.40 mg/lb], q 12 h). All successfully underwent surgical adrenal tumor extirpation. Thirty-eight dogs had pituitary-dependent hyperadrenocorticism (PDH); 15 dogs did not require a dose increase during the study, and at each of 4 reevaluations, 10 of 15, 13 of 15, 14 of 15, and 11 of 11 had a good response. Twenty-three dogs with PDH had their dose or frequency of trilostane administration increased during the study. Mean trilostane dosage at 1-year reevaluation in dogs with a good response was 1.7 mg/kg (0.8 mg/lb), twice daily, or 1.1 mg/kg, 3 times daily. At each of 4 reevaluations, 17 of 23, 14 of 23, 17 of 23, and 13 of 17 dogs with PDH had a good response. Five dogs became ill because of trilostane-induced adverse effects, but only 1 required hospitalization. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of initial lower doses of trilostane to dogs with NOH is effective. PMID- 21627508 TI - Diagnostic accuracy of using erythrocyte indices and polychromasia to identify regenerative anemia in dogs. AB - OBJECTIVE: To determine diagnostic accuracy of using erythrocyte indices and polychromasia to identify regenerative anemia in dogs. DESIGN: Retrospective and prospective cross-sectional study. ANIMALS: 4,521 anemic dogs. PROCEDURES: CBC results obtained between July 2002 and July 2008 by use of an automated laser based flow cytometric hematology analyzer from dogs with Hct values <= 35% were retrieved. Sensitivity, specificity, accuracy, and predictive values of using erythrocyte indices and polychromasia to identify regeneration were determined, with a reticulocyte count > 65,000 reticulocytes/MUL considered the gold standard. Similarly, 134 blood samples from anemic dogs were analyzed prospectively with an in-house electrical impedance analyzer. RESULTS: Of 4,387 dogs with samples analyzed retrospectively, 1,426 (32.5%) had regenerative anemia. Of these, 168 (11.8%) had macrocytic hypochromic anemia. High mean cell volume and low mean cell hemoglobin concentration had low sensitivity (11%), high specificity (98%), and moderate accuracy (70%) when used to identify regenerative anemia. Use of polychromasia alone had an accuracy of 77%, and use of polychromasia combined with a high RBC distribution width (RDW) had an accuracy of 79%. Results obtained with the in-house analyzer were similar. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that most regenerative anemias in dogs were not macrocytic hypochromic. Polychromasia, with or without high RDW, was a more accurate indicator than other erythrocyte indices of regenerative anemia. To avoid a false diagnosis of nonregenerative anemia, a blood smear should be evaluated in anemic dogs when a reticulocyte count is not available. PMID- 21627509 TI - Radiographic reference limits for cardiac width in peregrine falcons (Falco peregrinus). AB - OBJECTIVE: To determine radiographic reference limits for cardiac width (CW) in peregrine falcons. DESIGN: Retrospective cross-sectional study. Animals-100 peregrine falcons. PROCEDURES: CW, thoracic width (TW), and sternal width (SW) were measured at the level of maximum CW on ventrodorsal radiographs of 64 falcons. Ratios of CW:TW and CW:SW were compared between positive pressure insufflation (PPI) radiographs and standard radiographs in 6 falcons. A hypothesis for the most accurate predictor of CW was formulated and validated in a new set of 36 falcons. RESULTS: PPI radiographs had a significantly lower CW:TW ratio than did standard radiographs. The CW:SW ratio was unaffected. The 95% confidence interval for a regression-based predicted CW was 19% larger for any given TW, compared with SW. The difference between predicted and measured CW was significantly smaller with SW, compared with TW. Validation of these findings in a new data set of 36 falcons confirmed that prediction of CW when SW was used as the independent variable in a regression equation was superior to use of the CW:SW ratio or the CW:TW ratio and was superior to prediction of CW when TW was used as the independent variable in a regression equation. CONCLUSIONS AND CLINICAL RELEVANCE: Of the variables examined, SW seemed to be the most precise and most accurate predictor for CW in peregrine falcons. Thoracic width seemed inferior as a predictor for CW because TW was influenced by respiratory movements. The following formula can be used to predict the 95% confidence interval for CW in peregrine falcons: CW = 0.83SW + 0.37 +/- 0.16. PMID- 21627510 TI - Association of histologic tumor characteristics and size of surgical margins with clinical outcome after surgical removal of cutaneous mast cell tumors in dogs. AB - OBJECTIVE: To evaluate the relationship between width and depth of surgical margins, amount of edema within and around the tumor, and degree of demarcation between the tumor and surrounding tissues with the clinical outcome following surgical removal of cutaneous mast cell tumors (cMCTs) in dogs. DESIGN: Retrospective cohort study. ANIMALS: 100 dogs with 115 resectable cMCTs. PROCEDURES: Information about the dogs' clinical outcomes following cMCT removal was obtained from primary care veterinarians. Histologic sections of excised tumors were assessed retrospectively for tumor grade and measurement of the narrowest lateral and deep margins of nonneoplastic tissue excised with the tumors; edema within the tumor and surrounding tissues was assessed as minimal, moderate, or severe. Tumors were classified as poorly, moderately, or well demarcated on the basis of the degree of mast cell infiltration into the adjoining connective tissue. RESULTS: Following tumor excision (with no additional postsurgery treatment), 96 dogs had no local recurrence or metastatic disease for 27 to 31 months; 4 metastatic disease-related deaths (dogs with grade II or III tumors) occurred within 3 to 9 months. Histologically, mean lateral and deep surgical margins around the tumors were 8.9 and 5.3 mm, respectively. No recurrence of tumor or metastatic disease developed following excision with lateral margins >= 10 mm and deep margins >= 4 mm. Edema and degree of demarcation were not correlated with outcome. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that most grade I and II cMCTs in dogs can be successfully treated by complete surgical removal with margins smaller than those currently recommended. PMID- 21627511 TI - Osteoma of the oral and maxillofacial regions in cats: 7 cases (1999-2009). AB - Objective-To describe clinical features of oral and maxillofacial osteomas in cats. Design-Retrospective case series. Animals-7 cats with oral or maxillofacial osteoma or both. Procedures-Medical records were reviewed for information on signalment, history, clinical signs, physical examination findings, diagnostic imaging findings, results of serum biochemical analyses and histologic testing, surgical procedures performed, and perioperative complications. Outcome was determined on the basis of follow-up telephone interviews of owners. Results-Cats ranged from 1 to 23 years of age. Clinical signs were observed in 5 cats and were attributed to the presence of the mass. Diagnostic imaging (radiography and computed tomography) and histologic examination confirmed the diagnosis of osteoma. Three cats were euthanatized; 1 cat was treated by mandibulectomy, 1 was treated by maxillectomy, and 2 were treated by debulking. At the time of follow up at least 1 year after surgery, all 4 treated cats were alive, with owners reporting an acceptable quality of life. Conclusions and Clinical Relevance Osteoma of the oral and maxillofacial regions is an uncommon tumor in cats. Most cats are examined during an advanced stage of the disease, when treatment options may be limited. Although osteoma is a benign tumor, the recommendation is to perform a clinical evaluation, diagnostic imaging, biopsy, and treatment early in the disease process, when less invasive surgical approaches may be feasible. PMID- 21627512 TI - Multisystemic infection with an Acanthamoeba sp in a dog. AB - CASE DESCRIPTION-A 10-month-old Boxer was evaluated for fever and signs of cervical pain. CLINICAL FINDINGS-Physical examination revealed lethargy, fever, and mucopurulent ocular and preputial discharge. On neurologic examination, the gait was characterized by a short stride. The dog kept its head flexed and resisted movement of the neck, consistent with cervical pain. Clinicopathologic findings included neutrophilic leukocytosis, a left shift, and monocytosis. Cervical radiographs were unremarkable. Cerebrospinal fluid analysis revealed neutrophilic pleocytosis and high total protein content. On the basis of signalment, history, and clinicopathologic data, a diagnosis of steroid responsive meningitis-arteritis was made. TREATMENT AND OUTCOME: The dog was treated with prednisone (3.2 mg/kg [1.45 mg/lb], PO, q 24 h), for 3 weeks with limited response. Consequently, azathioprine (2 mg/kg [0.9 mg/lb], PO, q 24 h) was administered. Three weeks later, the dog was evaluated for tachypnea and lethargy. Complete blood count revealed leukopenia, neutropenia, and a left shift. Thoracic radiography revealed a diffuse bronchointerstitial pattern. The dog subsequently went into respiratory arrest and died. On histologic evaluation, amoebic organisms were observed in the lungs, kidneys, and meninges of the brain and spinal cord. A unique Acanthamoeba sp was identified by use of PCR assay. CLINICAL RELEVANCE: This dog developed systemic amoebic infection presumed to be secondary to immunosuppression. The development of secondary infection should be considered in animals undergoing immunosuppression for immune-mediated disease that develop clinical signs unrelated to the primary disease. Although uncommon, amoebic infection may develop in immunosuppressed animals. Use of a PCR assay for identification of Acanthamoeba spp may provide an antemortem diagnosis. PMID- 21627513 TI - Serial use of serologic assays and fecal PCR assays to aid in identification of subclinical Lawsonia intracellularis infection for targeted treatment of Thoroughbred foals and weanlings. AB - OBJECTIVE: To assess the serial use of serum immunoperoxidase monolayer assays (IPMAs) and fecal PCR assays, combined with other diagnostic methods, to identify subclinical Lawsonia intracellularis infections for targeted treatment of Thoroughbred foals and weanlings at farms in which the pathogen was endemic or nonendemic. DESIGN: Evaluation study. ANIMALS: 100 foals and weanlings (53 and 47 at farms in which L intracellularis was endemic and nonendemic, respectively). PROCEDURES: Serum was collected every 4 weeks and tested via IPMA, for antibodies against L intracellularis. Fecal samples were collected every 2 weeks and tested by use of an L intracellularis-specific PCR assay. When results for IPMAs or PCR assays were positive or clinical signs compatible with equine proliferative enteropathy (EPE) were detected, clinicopathologic testing was performed to determine treatment. RESULTS: No foals had positive results for the L intracellularis-specific IPMA until after weaning; 32 of 53 (60.4%) weanlings at the farm in which L intracellularis was endemic and 8 of 47 (170%) at the farm in which L intracellularis was nonendemic had positive IPMA results, whereas the number of weanlings that tested positive via fecal PCR assays at those farms was 6 and 0, respectively. Nineteen of 32 weanlings with positive IPMA results at the farm in which L intracellularis was endemic were treated for EPE; 5 of these had clinical signs of EPE. No weanlings at the nonendemic farm had clinical signs of or were treated for EPE. CONCLUSIONS AND CLINICAL RELEVANCE: IPMA appeared to be a useful means of identifying weanlings exposed to L intracellularis. PMID- 21627514 TI - Use of a novel intramedullary nail for femoral fracture repair in calves: 25 cases (2008-2009). AB - OBJECTIVE: To describe the use of a novel intramedullary interlocking nail for femoral fracture repair in newborn calves and outcome associated with this treatment modality. DESIGN: Retrospective case series. ANIMALS: 25 calves. PROCEDURES: Medical records from calves referred for femoral fracture repair between November 2008 and June 2009 were included. Signalment, clinical findings, and results of lameness examination and complete radiographic examination were recorded. Details of surgical technique, orthopedic fixation, associated complications, and outcome were also noted. RESULTS: 25 Charolais calves were included in the study. All fractures were closed and located in the diaphysis; 18 extended to the distal metaphysis, 1 was located proximally, and 6 were midshaft fractures. Fractures were localized to the left femur in 17 calves and to the right femur in 8 calves. An interlocking nail was used as the sole means of internal fixation in 16 calves and was reinforced in 9 calves. Two calves were euthanatized for reasons related to the surgery, and 4 died of unrelated reasons. Long-term prognosis was considered excellent for 15 calves and fair for 4 calves. Age, body weight, overall musculoskeletal strength at initial evaluation, affected limb, configuration and location of the fracture, need for reinforcement of the repair, and quality of the reduction had no significant association with positive outcome of the surgery. CONCLUSIONS AND CLINICAL RELEVANCE: The novel intramedullary interlocking nail used in the present study was associated with a good prognosis for surgical repair of femoral fractures in newborn calves regardless of the location of the fracture. PMID- 21627516 TI - Sole depth and weight-bearing characteristics of the palmar surface of the feet of feral horses and domestic Thoroughbreds. AB - OBJECTIVE: To determine solar load-bearing structures in the feet of feral horses and investigate morphological characteristics of the sole in feral horses and domestic Thoroughbreds. SAMPLE: Forelimbs from cadavers of 70 feral horses and 20 domestic Thoroughbreds in Australia. PROCEDURES: Left forefeet were obtained from 3 feral horse populations from habitats of soft substrate (SS [n = 10 horses]), hard substrate (HS [10]), and a combination of SS and HS (10) and loaded in vitro. Pressure distribution was measured with a pressure plate. Sole depth was measured at 12 points across the solar plane in feet obtained from feral horses from SS (n = 20 horses) and HS (20) habitats and domestic Thoroughbreds (20). RESULTS: Feet of feral horses from HS habitats loaded the periphery of the sole and hoof wall on a flat surface. Feral horses from HS or SS habitats had greater mean sole depth than did domestic Thoroughbreds. Sole depth was greatest peripherally and was correlated with the loading pattern. CONCLUSIONS AND CLINICAL RELEVANCE: The peripheral aspect of the sole in the feet of feral horses had a load-bearing function. Because of the robust nature of the tissue architecture, the hoof capsule of feral horses may be less flexible than that of typical domestic horses. The application of narrow-web horseshoes may not take full advantage of the load-bearing and force-dissipating properties of the peripheral aspect of the sole. Further studies are required to understand the effects of biomechanical stimulation on the adaptive responses of equine feet. PMID- 21627517 TI - Antinociceptive effects of nalbuphine hydrochloride in Hispaniolan Amazon parrots (Amazona ventralis). AB - OBJECTIVE: To evaluate the antinociceptive effects and duration of action of nalbuphine HCl administered IM on thermal thresholds in Hispaniolan Amazon parrots (Amazona ventralis). ANIMALS: 14 healthy adult Hispaniolan Amazon parrots of unknown sex. PROCEDURES: 3 doses of nalbuphine (12.5, 25, and 50 mg/kg, IM) and saline (0.9% NaCl) solution (control treatment) were evaluated in a blinded complete crossover experimental design by use of foot withdrawal threshold to a noxious thermal stimulus. Baseline data on thermal threshold were generated 1 hour before administration of nalbuphine or saline solution; thermal threshold measurements were obtained 0.5, 1.5, 3, and 6 hours after administration. RESULTS: Nalbuphine administered IM at 12.5 mg/kg significantly increased the thermal threshold (mean change, 2.4 degrees C), compared with results for the control treatment, and significantly changed thermal threshold for up to 3 hours, compared with baseline results (mean change, 2.6 degrees to 3.8 degrees C). Higher doses of nalbuphine did not significantly change thermal thresholds, compared with results for the control treatment, but had a significant effect, compared with baseline results, for up to 3 and 1.5 hours after administration, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Nalbuphine administered IM at 12.5 mg/kg significantly increased the foot withdrawal threshold to a thermal noxious stimulus in Hispaniolan Amazon parrots. Higher doses of nalbuphine did not result in significantly increased thermal thresholds or a longer duration of action and would be expected to result in less analgesic effect than lower doses. Further studies are needed to fully evaluate the analgesic effects of nalbuphine in psittacine species. PMID- 21627518 TI - Pharmacokinetics of nalbuphine hydrochloride after intravenous and intramuscular administration to Hispaniolan Amazon parrots (Amazona ventralis). AB - OBJECTIVE: To assess the pharmacokinetics of nalbuphine HCl after IV and IM administration to Hispaniolan Amazon parrots (Amazona ventralis). ANIMALS: 8 healthy adult Hispaniolan Amazon parrots of unknown sex. PROCEDURES: Nalbuphine HCl (12.5 mg/kg) was administered IV and IM to all birds in a complete randomized crossover study design; there was a washout period of 21 days between subsequent administrations. Plasma samples were obtained from blood collected at predetermined time points for measurement of nalbuphine concentration by use of liquid chromatography-tandem mass spectrometry. Pharmacokinetic parameters were estimated by use of computer software. RESULTS: Nalbuphine was rapidly eliminated with a terminal half-life of 0.33 hours and clearance of 69.95 mL/min/kg after IV administration and a half-life of 0.35 hours after IM administration. Volume of distribution was 2.01 L/kg after IV administration. The fraction of the dose absorbed was high (1.03) after IM administration. No adverse effects were detected in the parrots during the study. CONCLUSIONS AND CLINICAL RELEVANCE: In Hispaniolan Amazon parrots, nalbuphine appeared to have good bioavailability after IM administration and was rapidly cleared after IV and IM administration. Safety and analgesic efficacy of various nalbuphine treatment regimens in this species require further investigation to determine the potential for clinical palliation of signs of pain in psittacine species. PMID- 21627519 TI - Effect of topical vapocoolant spray on response to arthrocentesis and intravenous catheterization in unsedated horses. AB - OBJECTIVE: To assess the efficacy of a commercially available topical vapocoolant spray in reducing responses to arthrocentesis of the middle carpal (MC) and metacarpophalangeal (MCP) joints and jugular vein catheterization in unsedated horses. ANIMALS: 8 healthy research horses. PROCEDURES: Arthrocentesis of both MC and MCP joints and bilateral jugular vein catheterization were performed in each horse. Immediately prior to skin penetration, 1 randomly selected MC joint, MCP joint, and jugular vein were sprayed with a vapocoolant liquid (intervention product), and the contralateral MC joint, MCP joint, and jugular vein were sprayed with water (placebo). An observer blinded to the type of spray treatment used evaluated the horses' responses to needle or catheter placement procedures by use of a 6-point categorical scale and a 100-mm visual analog scale. RESULTS: Responses evaluated via the visual analog scale were significantly reduced after application of the intervention product, compared with responses after application of the placebo, for the MC and MCP joints; no difference in responses to jugular vein catheterization was detected between the 2 treatments. CONCLUSIONS AND CLINICAL RELEVANCE: Vapocoolant spray was safe and effective in reducing horses' responses to arthrocentesis. The use of such products prior to joint injections may reduce procedural nociception and pain anticipation in unsedated horses and may improve the safety of personnel performing such procedures. PMID- 21627520 TI - Accuracy of isoflurane, halothane, and sevoflurane vaporizers during high oxygen flow and at maximum vaporizer dial setting. AB - OBJECTIVE: To assess the accuracy of isoflurane, halothane, and sevoflurane vaporizers during high oxygen flow and at maximum dial settings at room temperature and to test sevoflurane vaporizers similarly during heating and at low-fill states. SAMPLE: 5 isoflurane, 5 halothane, and 5 sevoflurane vaporizers. PROCEDURES: Vaporizers were tested at an oxygen flow of 10 L/min and maximum dial settings for 15 minutes under various conditions. All 3 vaporizer types were filled and tested at room temperature (21 degrees to 23 degrees C). Filled sevoflurane vaporizers were wrapped with circulating hot water (42 degrees C) blankets for 2 hours and tested similarly, and near-empty sevoflurane vaporizers were tested similarly at room temperature. During each 15-minute test period, anesthetic agent concentration was measured at the common gas outlet with a portable refractometer and temperature of the vaporizer wall was measured with a thermistor. RESULTS: For each vaporizer type, anesthetic agent concentrations and vaporizer wall temperatures decreased during the 15-minute test period. Accuracy of isoflurane and halothane vaporizers remained within the recommended 20% (plus or minus) deviation from dial settings. Heated and room-temperature sevoflurane vaporizers were accurate to within 23% and 11.7% (plus or minus) of dial settings, respectively. Sevoflurane vaporizers at low-fill states performed similarly to vaporizers at full-fill states. CONCLUSIONS AND CLINICAL RELEVANCE: Under these study conditions, the isoflurane and halothane vaporizer models tested were accurate but the sevoflurane vaporizers were not. Sevoflurane vaporizer accuracy was not affected by fill state but may be improved with vaporizer heating; measurements of inspired anesthetic agent concentrations should be obtained during the use of heated vaporizers. PMID- 21627521 TI - Effect of perzinfotel and a proprietary phospholipase A(2) inhibitor on kinetic gait and subjective lameness scores in dogs with sodium urate-induced synovitis. AB - OBJECTIVE: To investigate the ability of perzinfotel (an N-methyl-d-aspartate receptor antagonist) and a proprietary phospholipase A(2) (PLA(2)) inhibitor to attenuate lameness in dogs with sodium urate (SU)-induced synovitis. ANIMALS: 8 adult dogs. PROCEDURES: A blinded 4-way crossover study was performed. Dogs received perzinfotel (10 mg/kg), a proprietary PLA(2) inhibitor (10 mg/kg), carprofen (4.4 mg/kg; positive control treatment), or no treatment (negative control treatment). On the fourth day after initiation of treatment, synovitis was induced via intra-articular injection of SU 1 hour before administration of the last treatment dose. Ground reaction forces were measured and clinical lameness evaluations were performed before (baseline [time 0]) and 2, 4, 6, 8, 12, and 25 hours after SU injection. There was a 21-day washout period between subsequent treatments. Data were analyzed via repeated-measures ANOVAs. RESULTS: Peak vertical force (PVF) and vertical impulse (VI) values for negative control and perzinfotel treatments were significantly lower at 2 and 4 hours, compared with baseline values. Values for PVF and VI for the PLA(2) inhibitor and positive control treatments did not differ from baseline values at any time points. Between-treatment comparisons revealed significantly higher PVF and VI values for the positive control treatment than for the negative control and perzinfotel treatments at 2 and 4 hours. Values for VI were higher for PLA(2) inhibitor treatment than for negative control treatment at 2 hours. CONCLUSIONS AND CLINICAL RELEVANCE: Perzinfotel did not significantly alter SU-induced lameness. The proprietary PLA(2) inhibitor attenuated lameness but not as completely as did carprofen. PMID- 21627522 TI - Plasma concentrations and behavioral, antinociceptive, and physiologic effects of methadone after intravenous and oral transmucosal administration in cats. AB - OBJECTIVE: To determine plasma concentrations and behavioral, antinociceptive, and physiologic effects of methadone administered via IV and oral transmucosal (OTM) routes in cats. ANIMALS: 8 healthy adult cats. PROCEDURES: Methadone was administered via IV (0.3 mg/kg) and OTM (0.6 mg/kg) routes to each cat in a balanced crossover design. On the days of drug administration, jugular catheters were placed in all cats under anesthesia; a cephalic catheter was also placed in cats that received methadone IV. Baseline measurements were obtained >= 90 minutes after extubation, and methadone was administered via the predetermined route. Heart and respiratory rates were measured; sedation, behavior, and antinociception were evaluated, and blood samples were collected for methadone concentration analysis at predetermined intervals for 24 hours after methadone administration. Data were summarized and evaluated statistically. RESULTS: Plasma concentrations of methadone were detected rapidly after administration via either route. Peak concentration was detected 2 hours after OTM administration and 10 minutes after IV administration. Mean +/- SD peak concentration was lower after OTM administration (81.2 +/- 14.5 ng/mL) than after IV administration (112.9 +/- 28.5 ng/mL). Sedation was greater and lasted longer after OTM administration. Antinociceptive effects were detected 10 minutes after administration in both groups; these persisted >= 2 hours after IV administration and >= 4 hours after OTM administration. CONCLUSIONS AND CLINICAL RELEVANCE: Despite lower mean peak plasma concentrations, duration of antinociceptive effects of methadone was longer after OTM administration than after IV administration. Methadone administered via either route may be useful for perioperative pain management in cats. PMID- 21627523 TI - Effects of extracorporeal shock wave therapy and polysulfated glycosaminoglycan treatment on subchondral bone, serum biomarkers, and synovial fluid biomarkers in horses with induced osteoarthritis. AB - OBJECTIVE: To evaluate effects of extracorporeal shock wave therapy (ESWT) and polysulfated glycosaminoglycan treatment (PSGAGT) on subchondral bone (SCB), serum biomarkers, and synovial fluid biomarkers in horses with induced osteoarthritis. ANIMALS: 24 healthy 2- to 3-year-old horses. PROCEDURES: An osteochondral fragment was created on the distal aspect of the radial carpal bone in 1 middle carpal joint of each horse. Horses were randomly allocated to receive local application of ESWT (days 14 and 28; n = 8), PSGAGT (IM, q 4 d for 28 days; 8), or a sham ESWT probe (placebo; days 14 and 28; 8). Serum biomarkers were measured every 7 days, and synovial fluid biomarkers were measured every 14 days. Bone density was measured by use of computed tomography on days 0 and 70, and microdamage and bone formation variables were compared among groups at the end of the study (day 70). RESULTS: There was no significant effect of ESWT or PSGAGT on any bone variable. Serum osteocalcin concentration was significantly greater in horses that received ESWT, compared with placebo-treated horses, and serum concentration of the C-terminal telopeptide of type I collagen was significantly higher in horses that received ESWT, compared with placebo- and PSGAG-treated horses. Concentrations of the synovial fluid epitope CS846 were significantly higher in joints with osteoarthritis treated with ESWT CONCLUSIONS AND CLINICAL RELEVANCE: Treatment of osteoarthritis with ESWT had no effect on SCB but did induce increases in serum biomarkers indicative of bone remodeling. Treatment of osteoarthritis with PSGAG had no effect on SCB or biomarkers. PMID- 21627524 TI - Impact of successive freezing-thawing cycles on 3-T magnetic resonance images of the digits of isolated equine limbs. AB - OBJECTIVE: To assess the impact of cycles of freezing and thawing on magnetic resonance (MR) images (obtained by use of a 3-T magnet) of equine feet examined ex vivo. SAMPLE: 9 forelimbs from 9 horse cadavers. PROCEDURES: 9 forefeet underwent MR imaging first at ambient temperature within 12 hours after the horses' death and then after each freezing-thawing cycle. Three digits underwent freezing and thawing (at 4 degrees C for 36 hours) 2 times, 3 digits underwent freezing and thawing (at 4 degrees C for 36 hours) once and rescanning after 24 hours at ambient temperature, and 3 digits underwent freezing and thawing at ambient temperature for 24 hours once. Images of the digits obtained prior to freezing were subjectively compared with images obtained after freezing and thawing. Changes in the signal-to-noise ratio between examinations were assessed. RESULTS: Overall image quality was considered unchanged except for the hoof capsule. Quantitative analysis revealed signal-to-noise ratio changes in bone marrow, soft tissues, and hoof capsule induced with both thawing processes. The signal-to-noise ratio in the synovial recess of the distal interphalangeal joint significantly increased as a result of thawing at 4 degrees C. CONCLUSIONS AND CLINICAL RELEVANCE: Although overall image quality was considered unchanged except for the hoof capsule, results suggested that changes induced in cadaver limbs following freezing and thawing, which are probably attributable both to modified and inhomogeneous temperature distribution and direct tissue damage, may alter the reliability of signal intensity in ex vivo MR examinations. PMID- 21627525 TI - Evaluation of experimentally induced injury to the superficial digital flexor tendon in horses by use of low-field magnetic resonance imaging and ultrasonography. AB - OBJECTIVE: To evaluate tendon injuries in horses over a 16-week period by use of ultrasonography and low-field magnetic resonance imaging (MRI). SAMPLE: Tendons of 8 young adult horses. PROCEDURES: The percentage of experimentally induced tendon injury was evaluated in cross section at the maximal area of injury by use of ultrasonography and MRI at 3, 4, 6, 8, and 16 weeks after collagenase injection. The MRI signal intensities and histologic characteristics of each tendon were determined at the same time points. RESULTS: At 4 weeks after collagenase injection, the area of maximal injury assessed on cross section was similar between ultrasonography and MRI. In lesions of > 4 weeks' duration, ultrasonography underestimated the area of maximal cross-sectional injury by approximately 18%, compared with results for MRI. Signal intensity of lesions on T1-weighted images was the most hyperintense of all the sequences, lesions on short tau inversion recovery images were slightly less hyperintense, and T2 weighted images were the most hypointense. Signal intensity of tendon lesions was significantly higher than the signal intensity for the unaltered deep digital flexor tendon. Histologically, there was a decrease in proteoglycan content, an increase in collagen content, and minimal change in fiber alignment during the 16 weeks of the study. CONCLUSIONS AND CLINICAL RELEVANCE: Ultrasonography may underestimate the extent of tendon damage in tendons with long-term injury. Low field MRI provided a more sensitive technique for evaluation of tendon injury and should be considered in horses with tendinitis of > 4 weeks' duration. PMID- 21627526 TI - Comparison of two commercial ovine Campylobacter vaccines and an experimental bacterin in guinea pigs inoculated with Campylobacter jejuni. AB - OBJECTIVE: To compare efficacy of 2 commercial ovine Campylobacter vaccines and an experimental bacterin in guinea pigs following IP inoculation with Campylobacter jejuni IA3902. ANIMALS: 51 female guinea pigs. PROCEDURES: Pregnant and nonpregnant animals were randomly assigned to 1 of 4 treatment groups and administered a commercial Campylobacter vaccine labeled for prevention of campylobacteriosis in sheep via two 5-mL doses 14 days apart (vaccine A; n = 13), another labeled for prevention of campylobacteriosis via two 2-mL doses (vaccine B; 12), an experimental bacterin prepared from the challenge strain (12), or a sham vaccine (14). Ten days later, animals were challenged IP with C jejuni IA3902; 48 hours later, animals were euthanized, complete necropsy was performed, and blood and tissue samples were obtained for bacteriologic culture. RESULTS: Administration of vaccine B or the experimental bacterin, but not vaccine A, significantly reduced 48-hour infection rates versus administration of the sham vaccine. A significantly reduced 48-hour infection rate was associated with administration of vaccine B independent of pregnancy status. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of vaccine B significantly reduced infection in guinea pigs challenged with C jejuni IA3902, similar to a homologous bacterin. Results suggested that vaccine B or an autogenous product may be effective in controlling ovine campylobacteriosis caused by this emergent abortifacient strain. Bacteriologic culture of blood, liver, bile, and uterus in nonpregnant guinea pigs 48 hours after inoculation may be a useful screening tool for comparing efficacy of C jejuni vaccines. PMID- 21627527 TI - Shedding of chlamydiae in relation to titers of serum chlamydiae-specific antibodies and serum concentrations of two acute-phase proteins in cats without conjunctivitis. AB - OBJECTIVE: To investigate shedding of chlamydiae from conjunctiva and genital tracts of cats without clinical signs of conjunctivitis or other infectious disease in relation to their titers of serum antibodies against chlamydiae and to serum amyloid A (SAA) and serum alpha(1)-acid glycoprotein (AGP) concentrations. ANIMALS: 62 healthy cats. PROCEDURES: Serum from each cat was analyzed for antibodies against chlamydiae and for SAA and AGP concentrations. Swab samples from the conjunctival sac and genital tract were analyzed with a real-time PCR assay for Chlamydiaceae. RESULTS: 4 of 8 of cats with high antibody titers (ie, 1,600) shed chlamydiae, but only from the conjunctiva. Chlamydiae could not be detected in samples from cats with lower antibody titers nor from any genital tract samples. In cats with antibody titers of 1,600, mean +/- SD SAA concentration was significantly higher when chlamydiae were detected in conjunctival swab samples (3.9 +/- 1.0 mg/L) than when no chlamydiae were detected (1.4 +/- 1.0 mg/L). However, SAA concentration was greater than the limit for an acute-phase response in only one of those cats. There was no significant difference in serum AGP concentrations between cats with high titers that were or were not shedding chlamydiae. Nine of 30 (30%) cats (5 with and 4 without detectable serum antibodies against chlamydiae) that had been mated developed reproductive disorders. CONCLUSIONS AND CLINICAL RELEVANCE: Clinically normal cats with high chlamydiae-specific antibody titers can shed and thus transmit chlamydiae. Venereal spread from cats without clinical signs of infection is likely not common. PMID- 21627528 TI - Evaluation of the ability of two transfection reagents to deliver small interfering RNA molecules to equine and guinea pig cartilage in vitro. AB - OBJECTIVE: To evaluate 2 commercially available transfection reagents for transfection efficiency and distribution of small interfering RNA (siRNA) molecules to chondrocytes in monolayer cultures and full-thickness cartilage explants from guinea pigs and horses. SAMPLE: Cartilage explants from 5 one-month old and 3 adult guinea pigs and 5 adult clinically normal horses. PROCEDURES: Monolayer chondrocytes and uniform cartilage explants were exposed to 1 of 2 siRNA transfection complexes according to manufacturers' protocols (1MUM [1*]). Additionally, monolayer chondrocytes were exposed to 2* the suggested amount of a proprietary siRNA molecule. Full-thickness cartilage explants were treated with 1* (1MUM), 2* (2MUM), and 4* (4MUM) or 1* (0.13MUM), 4* (0.52MUM), and 8* (1.04MUM) the recommended concentrations of the proprietary siRNA and the cationic liposome siRNA, respectively, in equivalent media volumes. Use of fluorescent siRNA duplexes allowed quantification of transfected cells via flow cytometry and direct visualization of the depth and distribution of in situ transfection via fluorescent microscopy. RESULTS: With both transfection reagents, > 90% of monolayer chondrocytes were transfected. In explants, only use of the proprietary molecule achieved > 50% transfection efficiency, whereas use of the cationic liposome achieved < 20%. Only the proprietary molecule-treated cartilage consistently contained fluorescent cells throughout all zones; the cationic liposome-transfected chondrocytes were restricted to explant surfaces. CONCLUSIONS AND CLINICAL RELEVANCE: Robust transfection of chondrocytes in monolayer was achieved with both reagents, but only use of the proprietary molecule attained effective full-thickness transfection of explants that may allow relevant transcript reduction via RNAi. PMID- 21627529 TI - Accuracy of pressure plate kinetic asymmetry indices and their correlation with visual gait assessment scores in lame and nonlame dogs. AB - OBJECTIVE: To determine the accuracy of pressure plate kinetic asymmetry indices (ASIs) for diagnosis of unilateral hind limb lameness in dogs and their correlation with visual gait assessment (VGA) scores. ANIMALS: 9 healthy dogs and 16 dogs with previously diagnosed unilateral rupture of the cranial cruciate ligament and concurrent unilateral hind limb lameness. PROCEDURES: Dogs were walked over a pressure plate to determine paw contact area (PCA), peak vertical pressure (PVP), peak vertical force (PVF), and vertical impulse (VI) of both hind limbs. An ASI was calculated for each gait variable. Simultaneously, gait was assessed visually and scored by use of a numeric rating scale (0 to 10). The ASI of each variable was tested for its usefulness in discrimination between lame and nonlame dogs and for correlation with VGA scores. RESULTS: Sensitivity and specificity of ASIs to discriminate between lame and nonlame dogs were excellent for PVF, VI, and PCA; these values were substantially lower for ASI of PVP. Cutoff values to discriminate between lame and nonlame dogs were determined by use of ASIs for PVF, VI, and PCA; however, this could not be done for ASI of PVP. Correlations between ASIs of PVF, VI, and PCA and VGA scores were higher than correlation between the ASIs of PVP and VGA scores. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that ASIs of PVF and VI determined via analysis of pressure plate measurements were reliable indicators of clinical lameness in dogs, but the ASI of PVP was not. The ASI of PCA is an interesting new variable for assessment of limb loading symmetry. PMID- 21627530 TI - Evaluation of four drugs for inhibition of paracentesis-induced blood-aqueous humor barrier breakdown in cats. AB - OBJECTIVE: To compare inhibitory effects of topically applied 1% prednisolone acetate suspension, 0.03% flurbiprofen solution, 0.1% dexamethasone suspension, and 0.1% diclofenac solution on paracentesis-induced blood-aqueous barrier breakdown in cats. ANIMALS: 9 healthy cats. PROCEDURES: Paracentesis of the anterior chamber was performed in both eyes of each cat. One eye of each cat was treated with a topically administered anti-inflammatory medication (1% prednisolone [n = 7 cats], 0.03% flurbiprofen [7], 0.1% dexamethasone [9], or 0.1% diclofenac [8]) immediately following paracentesis and at 6, 10, and 24 hours after paracentesis. The contralateral untreated eye served as the control eye. Each cat had a 6-day washout period between experimental drugs. Breakdown of the blood-aqueous barrier was quantified by use of laser flaremetry. RESULTS: Topical administration of 1% prednisolone significantly reduced aqueous humor flare at 4, 8, and 26 hours after paracentesis. Topical administration of 0.1% diclofenac significantly reduced aqueous humor flare at 8 and 26 hours after paracentesis. Topical administration of 0.1% dexamethasone and 0.03% flurbiprofen did not significantly decrease flare at any time point. There were significant differences in intraocular pressures between NSAID-treated eyes and untreated contralateral eyes. CONCLUSIONS AND CLINICAL RELEVANCE: Topical administration of 1% prednisolone and 0.1% diclofenac significantly reduced intraocular inflammation in cats with paracentesis-induced uveitis. Topical administration of 1% prednisolone or 0.1% diclofenac may be appropriate choices when treating cats with anterior uveitis. Topical administration of diclofenac and flurbiprofen should be used with caution in cats with a history of ocular hypertension. PMID- 21627531 TI - Development and validation of a sandwich ELISA for use in measuring concentrations of canine surfactant protein A in serum of dogs. AB - OBJECTIVE: To develop and evaluate a sandwich ELISA incorporating rabbit antiserum specific for canine surfactant protein A (SP-A) for use in measuring concentrations of SP-A in serum of dogs. SAMPLE: Serum samples obtained from 6 healthy dogs and 3 dogs with pulmonary disease. PROCEDURES: Rabbit antiserum was prepared against purified canine SP-A. The IgG fraction was isolated via protein G affinity chromatography and was then biotinylated. The sandwich ELISA was performed by use of anti-SP-A antibody (IgG) preabsorbed with sera from healthy dogs. Validity of the ELISA was confirmed by determination of the detection limit, precision, reproducibility, and accuracy. Serum SP-A concentrations were measured in 6 healthy dogs and 3 dogs with pulmonary disease. RESULTS: Detection limit of the ELISA was 2.0 ng/mL. Within- and between-assay coefficients of variation ranged from 3.8% to 14.1% and from 15.5% to 35.6%, respectively. The observed-to-expected recovery ratio ranged from 77.1% to 89.9%. Serum SP-A concentrations measured by use of the ELISA were <= 2.3 ng/mL in the 6 healthy dogs, 25.6 ng/mL in a dog with severe cardiac pulmonary edema, 8.3 ng/mL in a dog with pneumonia, and 10.1 ng/mL in a dog with lung lobe torsion. CONCLUSIONS AND CLINICAL RELEVANCE: The sandwich ELISA was found to be useful for measuring purified canine SP-A concentrations and canine SP-A concentrations in serum samples. The ELISA was precise, reproducible, and accurate. The ELISA may be beneficial in assessing serum concentrations of canine SP-A as a potential biomarker of pulmonary diseases in dogs. PMID- 21627532 TI - Evaluation of variations in bronchoalveolar lavage fluid in horses with recurrent airway obstruction. AB - OBJECTIVE: To determine variations in cytologic counts of bronchoalveolar lavage (BAL) fluid attributable to month of collection, first and second aliquots, and left and right lung sites in horses with recurrent airway obstruction (RAO). ANIMALS: 5 horses with RAO and 5 healthy horses without respiratory tract disease. PROCEDURES: Horses were housed in a stable for 5 months prior to and throughout the study. Bronchoalveolar lavage fluid was collected from the right and left lung of each horse 3 times at monthly intervals (February, March, and April). Each BAL fluid collection was performed by use of 2 incremental instillations of 250 mL of isotonic saline (0.9% NaCl) solution in the same bronchial site. Analysis of BAL fluid included volume of BAL fluid recovered, a CBC, and differential cytologic counts. RESULTS: Volume of BAL fluid recovered and cytologic counts did not differ in horses with RAO across time or between right and left lungs, except for the number of mast cells. Horses with RAO had significantly lower volumes of BAL fluid recovered, significantly lower percentages of macrophages and lymphocytes, and significantly higher percentages of neutrophils than did healthy horses. Despite individual variation, all horses with RAO had > 25% neutrophils throughout the study period. CONCLUSIONS AND CLINICAL RELEVANCE: Despite variation among horses, BAL fluid cytologic counts were repeatable over short and long periods and samples can be used for longitudinal studies as a diagnostic tool of pulmonary inflammation in horses with RAO. PMID- 21627533 TI - Effects of oral administration of N-acetyl-d-glucosamine on plasma and urine concentrations of glycosaminoglycans in cats with idiopathic cystitis. AB - OBJECTIVE: To determine the effects of once-daily oral administration of N-acetyl d-glucosamine (NAG) on plasma and urine glycosaminoglycan (GAG) concentrations in cats with idiopathic cystitis (IC). ANIMALS: 19 cats with IC and 10 clinically normal cats. PROCEDURES: Cats with IC were randomly assigned to receive 250 mg of NAG in capsule form orally once daily for 28 days (n = 12) or a placebo (capsule containing cellulose) orally once daily for the same period (7). In cats with IC, plasma and urine GAG concentrations and urine creatinine concentration were measured on days 0 (immediately before first dose), 7, 14, 21, 28, and 56. For purposes of comparison, those variables were measured in 10 clinically normal cats on day 0. RESULTS: Mean +/- SEM urine GAG-to-creatinine concentration ratios (day 0 data) for cats with IC and clinically normal cats differed significantly (3.11 +/- 0.62 MUg/mL and 14.23 +/- 3.47 MUg/mL, respectively). For cats with IC, mean plasma GAG concentration in NAG-treated cats (39.96 +/- 5.34 MUg/mL) was higher than that in placebo-treated cats (24.20 +/- 3.35 MUg/mL) on day 21. In the NAG-treated cats, plasma GAG concentration on days 21 (39.96 +/- 5.34 MUg/mL) and 28 (39.91 +/- 6.74 MUg/mL) differed significantly from the day 0 concentration (27.46 +/- 3.90MUg/mL). CONCLUSIONS AND CLINICAL RELEVANCE: Cats with IC have lower urinary GAG-to-creatinine concentration ratios than did clinically normal cats. Administration of NAG (250 mg, PO, q 24 h) significantly increased plasma GAG concentrations in cats with IC after 21 days of treatment. PMID- 21627534 TI - Generation of artificial channels by multimerization of beta-strands from natural porin. AB - General diffusion porins are passive transmembrane channels. We have explored the possibility to create artificial nanopores starting from natural beta-barrel structures. Structural elements of bacterial porins were used to build a series of artificial nanopores. The basic module was selected by multi-alignment of general diffusion porins. The sequence corresponded to a highly conserved motif containing two beta-strands, which was obtained from Escherichia coli OmpF. Dimeric to octameric repeats were obtained through cDNA recombinant technology. The hexameric repeat was used to test its properties. This protein was expressed, purified and reconstituted in the planar bilayer membranes. It was able to form channels in membranes with a conductance of 300 pS in 150 mm KCl and did not show any relevant voltage-dependence. PMID- 21627535 TI - Mesothelial cells activate the plasma kallikrein-kinin system during pleural inflammation. AB - Abstract Pleural inflammation underlies the formation of most exudative pleural effusions and the plasma kallikrein-kinin system (KKS) is known to contribute. Mesothelial cells are the predominant cell type in the pleural cavity, but their potential role in plasma KKS activation and BK production has not been studied. Bradykinin concentrations were higher in pleural fluids than the corresponding serum samples in patients with a variety of diseases. Bradykinin concentrations did not correlate with disease diagnosis, but were elevated in exudative effusions. It was demonstrated, using a range of primary and transformed mesothelial and mesothelioma cell lines, that cells assembled high molecular weight kininogen and plasma prekallikrein to liberate bradykinin, a process inhibited by novobiocin, a heat shock protein 90 (HSP90) inhibitor, cysteine, bradykinin and protamine sulphate. Of the common plasma prekallikrein activators, mesothelial cells expressed HSP90, but not prolylcarboxypeptidase or Factor XII. Calcium mobilisation was induced in some mesothelium-derived cell lines by bradykinin. Des-Arg(9)-bradykinin was inactive, indicating that mesothelial cells are responsive to bradykinin, mediated via the bradykinin receptor subtype 2. In summary, pleural mesothelial cells support the assembly and activation of the plasma KKS by a mechanism dependent on HSP90, and may contribute to KKS-mediated inflammation in pleural disease. PMID- 21627536 TI - Impact of aryl hydrocarbon receptor (AhR) knockdown on cell cycle progression in human HaCaT keratinocytes. AB - Abstract While activation of the aryl hydrocarbon receptor (AhR) by exogenous ligands is well investigated, its physiological function is less understood. By extending research in AhR biology, evidence appeared that the receptor generally plays an important role in cell physiology. In keratinocytes, little is known about endogenous functions of the AhR. In order to expand this knowledge, we analyzed the impact of AhR knockdown on cell cycle progression in HaCaT cells and showed that proliferation of siAhR HaCaT cells was significantly decreased. In line with that result, western blot analysis revealed that protein level of the cyclin dependent kinase inhibitor p27(KIP1) was increased, whereas protein level of the cyclin dependent kinase (CDK) 2 was reduced. CDK4 and CDK6 protein levels remained unchanged, whereas protein level of the retinoblastoma protein (pRB) was reduced. By measuring ethoxyresorufin-O-deethylase (EROD) activity we showed that endogenous cytochrome P450 1 (CYP1), especially CYP1A1 is required for normal cell cycle in HaCaT cells, as well. To the best of our knowledge, we provide evidence for the first time in human skin cells, that in the absence of exogenous ligands, the AhR promotes cell cycle progression in HaCaT cells and one can speculate that this is the physiological function of this receptor in keratinocytes. PMID- 21627537 TI - Novel neutrophil inhibitory factor homologue in the buccal gland secretion of Lampetra japonica. AB - Abstract To identify the functional gene fragment, a neutrophil inhibitory factor (NIF) like protein was found in the buccal gland of Lampetra japonica, suggesting that this related lamprey protein represents a novel class of integrin receptor antagonists. The recombinant Lampetra japonica-NIF like (rLj-NIF) was identified by SDS-PAGE and purified by using His.Bind affinity chromatography. Effect of rLj NIF on neutrophil migration suggested that rLj-NIF can act as a neutrophil inhibitory factor. Besides that, oxidative burst activity of neutriphil was tested by flow cytometry using dihydrorhodamine (DHR123) as a fluorogenic substrate, and the data suggested that the mean fluorescence intensity significantly decreased compared with positive controls (p<0.01). All above results indicated that rLj-NIF could also prevent the binding of beta2 integrins to the surface of PMN and its FITC-labeled monoclonal antibodies (p<0.05). These data suggest that Lampetra japonica-NIF like protein is secreted by the stage of the parasite at the site of attachment. rLj-NIF plays an essential role in physiological reaction of neutrophil by a novel class of beta2 integrin receptor antagonists. The activity of immunosuppressant of L. japonica-NIF could have potential medicinal value in anti-inflammation and therapy of autoimmune diseases. PMID- 21627538 TI - Bilateral renal infarctions complicating fibromuscular dysplasia of renal arteries in a young male. AB - Fibromuscular dysplasia (FMD) is an uncommon disorder, accounting for less than 10% of cases of renal artery stenosis, and typically presenting with hypertension in young women. This article reports the case of a previously healthy 37-year-old man presenting with acute-onset, severe, bilateral flank pain. Initially treated for ureteral colic and urinary tract infection, he was transferred to the nephrology clinic upon recognition of a rising serum creatinine. He was found to have FMD of bilateral renal arteries with a stenotic pattern on the right side and a dissecting aneurysm on the left side with resultant infarctions in both kidneys. On the basis of negative serological markers of vasculitis, a diagnosis of FMD complicated by bilateral renal infarctions was made. A stent was placed to the right stenotic renal artery, which resulted in sufficient lumen patency. No invasive procedure was performed on the other side owing to the complexity of the lesion. After 2.5 years of follow-up, the patient remained in good condition with normal renal function and adequate blood pressure control with dual antihypertensive therapy. Renal infarction complicating FMD of renal arteries is rare in the literature, with most of the cases having causative cardiovascular risk factors including coagulopathy, ischaemic heart disease, atrial fibrillation or structural cardiac abnormalities, none of which was present in this case. In conclusion, FMD may occur in atypical asymmetric presentations causing renal infarctions in both kidneys. Radiological interventions in such cases should focus on stabilizing renal lesions and renal function. PMID- 21627539 TI - Harmonization of growth hormone measurements with different immunoassays by data adjustment. AB - BACKGROUND: The aim of our study was to evaluate the between-assay variability of commercially available immunoassays for the measurement of human growth hormone (hGH). In addition, we asked whether the comparability of the diagnosis of childhood onset growth hormone deficiency could be improved by adjusting hGH results by statistical methods, such as linear regression, conversion factors, and quantile transformation. METHODS: In archived sera from 312 children and adolescents (age: 17 days-17 years) hGH values between 0.01 and 16.5 ng/mL were determined by using the following immunoassays: AutoDELFIA (PerkinElmer), BC-IRMA (Beckman-Coulter), ELISA (Mediagnost), IMMULITE 2000 (Siemens), iSYS (IDS), Liaison (DiaSorin), UniCel DxI 800 Access (BeckmanCoulter) and "In house"-RIA (Tubingen). RESULTS: The assays differed in median hGH concentrations by as much as 5.44 ng/mL (Immulite), and as little as 2.67 ng/mL (BC-IRMA). The mean difference between assays ranged from 0.35 to 2.71 ng/mL, whereas several samples displayed differences up to 11.4 ng/mL. The best correlation (r=0.992) was found between AutoDELFIA and Liasion, the lowest (r=0.864) was between an in-house RIA and iSYS. The between-assay CV (mean +/- SD) of values within the cut-off range was 24.3% +/- 7.4%, resulting in an assay-dependent diagnosis of growth hormone deficiency (GHD) in more than 27% of patients. Yet, adjustment of this data by linear regression or a conversion factor reduced the CV below 14%, and the ratio of assay-dependent diagnoses below 8%. Using quantile transformation, the CV and ratio were reduced to 11.4% and <1%, respectively. CONCLUSIONS: hGH measurements using different assays vary significantly. Linear regression, conversion factors, or particularly quantile transformation are useful tools to improve comparability in the diagnostic procedure for the confirmation of GHD in childhood and adolescence. PMID- 21627540 TI - The function of oxalic acid in the human metabolism. AB - Biochemical reactions in cells which involve oxalic acid are described. It is shown that this compound is required for the formation of uracil and orotic acid. The former is a component of RNA which is common to all cells in the human metabolism. On the basis of the biochemical reactions described a possible treatment to relieve the effects of calcium oxalate renal calculi whose origin is related to the metabolic concentration of oxalic acid is proposed. PMID- 21627541 TI - High resolution melting analysis to genotype the most common variants in the HFE gene. AB - BACKGROUND: High resolution melting (HRM) analysis of PCR amplicons was recently introduced as a closed-tube, rapid, and inexpensive method of genotyping. This study evaluated this system as an option for detecting the three most common mutations in the HFE gene (C282Y, H63D, S65C), accounting for the main form of hereditary haemochromatosis. METHODS: Ninety samples, previously screened by direct sequencing, and 27 controls were used. The analysis were performed on the Rotor Gene Q, using the commercial HRM mix containing the Eva Green dye (Qiagen). Specific primers allowed the amplification of the regions of interest in the HFE gene. Following amplification, a HRM analysis was conducted to detect DNA variants. The thermal denaturation profiles of the samples were compared with those of the controls. RESULTS: One hundred percent of heterozygous and homozygous samples were readily identified. Heterozygotes were easily identified because heteroduplexes altered the shape of the melting curves, but significant differences were also present in the melting curves of the homozygous carries compared with those of the wild-type subjects. CONCLUSIONS: HRM analysis is an appealing technology for HFE gene screening. It is a robust technique that can be widely adopted in diagnostic laboratories to facilitate gene mutation screening. PMID- 21627543 TI - Social inequality in oral health-related quality-of-life, OHRQoL, at early older age: evidence from a prospective cohort study. AB - OBJECTIVES: This study aimed to assess the long-term effect of socio-behavioral characteristics on oral impacts on daily performances (OIDP) at age 65 and the influence on OIDP at age 65 of changes in socio-behavioral characteristics between age 50 and 65. METHODS: In 1992, all 50-year-olds in two counties of Sweden were invited to participate in a longitudinal survey. A total of 6346 responded and, of those, 4143 completed follow-ups at ages 55, 60 and 65 years. RESULTS: At age 65, 26.9% had oral impacts. Subjects being immigrants, being unmarried, reporting economic barriers, bad general health, bad quality dental care, less than annual dental attendance, limited social network and smoking at age 50 experienced oral impacts at age 65 more frequently than their counterparts in the opposite groups. Compared to the stable advantaged groups, stable disadvantaged groups regarding marital status, health status, smoking and reported quality of care had increased ORs for oral impacts. Compared to the stable advantaged groups, those who experienced deterioration with respect to health status, dental attendance and quality of oral health care and those who quitted smoking had increased ORs for oral impacts. CONCLUSIONS: Disadvantaged socio-behavioral condition at age 50 had a long lasting detrimental effect on OHRQoL at age 65. Deterioration in socio-behavioral circumstances was associated with increased oral impacts. Early protection against the effect of socio behavioral adversity by imposing economic barriers, ensure provision of high quality care and by promotion of healthy lifestyles seems to have the potential to reduce oral impacts at older ages. PMID- 21627544 TI - Targeted biological therapies for pain. AB - INTRODUCTION: Chronic pain is often resistant to currently used drugs. The effect of these is frequently self-limiting, with increasing level of side effects caused by increased doses. Biological pain therapies provide a means to target molecules to specific types of neural cells in spatially limited areas. Targeted biological therapies utilize agents acting at specific sites, or virus or cell vectors allowing expression and secretion of transgenic substances in small anatomical compartments. Biological approaches to treatment of chronic pain may be able to provide greater analgesic efficacy, avoiding many of the limitations associated with current analgesics. AREAS COVERED: The most important targets and tools for biological therapy of pain. Basic approaches, preclinical trials and the clinical studies successfully completed. The rationale and tools for biological therapies. EXPERT OPINION: Biological therapy of pain holds great promise and is rapidly developing. Despite the significant numbers of preclinical studies in the last two decades only a single biological agent, the cone snail toxin ziconotide, has been advanced through all stages and licensed for clinical use. Biological therapy of pain is thus here to stay, but will need more substantial proof of efficacy and safety before being widely accepted and routinely used. PMID- 21627545 TI - Cost and clinical consequences of stress/single-photon emission computed tomography testing with specific medications. AB - BACKGROUND: We identified the cost of care and clinical events using dipyridamole versus adenosine in pharmacological stress (ST)/single-photon-emission computed tomography (SPECT) myocardial perfusion testing. METHODS: Commercial health plan members received adenosine or dipyridamole as an adjunct to ST/SPECT testing during the period January 1, 2006, through November 30, 2008. Propensity score matching techniques were used to compare risk-adjusted, test-related complications, symptoms and costs. RESULTS: A total of 12,351 patients underwent ST/SPECT testing with dipyridamole and 59,969 with adenosine. Risk-adjusted outcomes analysis showed that patients receiving dipyridamole had a higher number of emergency room (ER) visits (0.65 vs. 0.23%, p<0.001) and angina pectoris episodes (7.11 vs. 6.01%, p<0.001). The likelihood of shortness of breath was significantly higher (6.63 vs. 5.77%, p<0.001) in the adenosine group. One-day risk-adjusted, office-visit, outpatient hospital and other utilization costs for same day ST/SPECT testing were higher for the adenosine group. Risk-adjusted ER visit costs were higher for the dipyridamole group ($1276 vs. $1095, p<0.001). LIMITATIONS: First the presence of a claim for a filled prescription does not indicate that the medication was consumed or taken as prescribed. Second, medications filled over-the-counter or provided as samples will not be observed in the claims data. Third, presence of diagnosis codes on medical claims are not positive presence of disease, as diagnosis codes may be incorrectly coded or included as rule-out criteria rather than actual disease. Finally, certain information is not readily available in claims data that could have an effect on study outcomes, such as certain clinical and disease-specific parameters. CONCLUSIONS: Differences in complications and symptoms may help identify a better tolerated vasodilator drug (VD) for use in pharmacologic stress testing based on a patient's history and symptoms. Implementation of a data-based strategy for the selection of the most appropriate stress-testing adjunctive agent may be a cost effective step for institutions and health plans. PMID- 21627546 TI - Toxicological considerations for intravitreal drugs. AB - INTRODUCTION: Intravitreal injections are a very common procedure and are the most effective route of drug delivery to the retina. There are currently several drugs available and even more are in development; therefore, safety is a very important concern. AREAS COVERED: The toxicological considerations of the most common drugs used for intravitreal pharmacotherapy such as anti-VEGFs, corticosteroids and antibiotics. Emerging agents such as anti-TNFs, VEGF-trap and kinase inhibitors are also discussed. An assessment of the efficacy and safety issues of the most relevant drugs including bevacizumab, ranibizumab and triamcinolone is presented. EXPERT OPINION: The toxicology and safety profiles are available for several drugs that are either in use or will be available for intravitreal injections. Retinal pharmacotherapy is very effective for different retinal diseases; however safety is a very important issue when intravitreal injections are applied and the possibility of retinal toxicity should always be kept in mind. Bevacizumab and ranibizumab are effective for the therapy of wet age-related macular degeneration and macular edema, while triamcinolone remains an alternative agent to treat secondary macular edema. It is important, as some of these drugs will be used for extended periods of time, that their long-term toxicological effects are better understood. PMID- 21627547 TI - Obstetric outcomes in pregnant women with diabetes versus hypertensive disorders versus both. AB - OBJECTIVE: To compare obstetrical outcomes in pregnant women with diabetes versus hypertensive disorders versus both. METHODS: One million patients in the New Jersey Database were analyzed. Of which 6.91% had hypertension, 4.79% had diabetes, and 0.91% had both. Information was derived from a perinatal linked data-set provided by the Maternal Child Health Epidemiology (MCH Epi) Program in the New Jersey Department of Health and Senior Services. Linking of electronic birth certificates, hospital discharge records for mother and newborn, and infant death certificates for all infants born in New Jersey between the years 1997 and 2005 created the data-set. RESULTS: Coexistence of hypertension and diabetes increased with advancing maternal age (OR 3.41; CI 3.12-3.72). Among ethnic groups, diabetes was more common in Asians (OR 2.92; CI 2.84-3.00), while hypertension was more common in Blacks (OR 1.49; CI 1.46-1.53). Blacks followed by Asians had a higher risk of being in the combined category. Induction of labor (OR 4.16; CI 3.96-4.38), shoulder dystocia (OR 2.56; CI 2.05-3.19), operative vaginal delivery (OR 3.92; CI 3.29-4.66), cesarean deliveries with no trial of labor (OR 2.54; CI 2.40-2.69) as well as with failed trial of labor (OR 4.09; CI 3.88-4.31) were more common in the combined group. Neonatal outcomes were poor in the combined category, with high rate of preterm deliveries, neonatal intensive care unit (NICU) admissions (OR 2.14; CI 2.01-2.28), neonatal seizures (OR 2.30; CI 1.31-4.04), low 5-min APGAR scores (OR 1.78; CI 1.57-2.01), and longer NICU stay (OR 2.30; CI 2.15-2.47). CONCLUSIONS: Coexistence of hypertension and diabetes was associated with worse obstetric and neonatal outcomes than either alone. This should be emphasized to mothers during prenatal counseling. Further research should focus on interventions to improve morbidity in the combined category. PMID- 21627548 TI - Effect of leptin treatment on neonatal hypoxic-ischemic brain injury. AB - OBJECTIVE: The aim of this study was to investigate the possible neuroprotective and ameliorating effects of leptin treatment in hypoxic-ischemic injury induced neuronal cell death. METHODS: Experimental groups in the study were: sham operated group, leptin treated hypoxia-ischemia group, and vehicle treated hypoxia-ischemia group. In hypoxia-ischemia group, left common carotid artery was ligated permanently on the seventh postnatal day. Two hours after the procedure, hypoxia (92% nitrogen and 8% oxygen) was applied for 2.5 h. Leptin treatment was injected (intraperitoneally; i.p.) as a single dose immediately after the hypoxia period. Neuronal cell death, neuronal density, and leptin levels were evaluated in both hemispheres 72 h after the hypoxic-ischemic insult. RESULTS: Compared with the hypoxic-ischemia group, the mean leptin levels were higher in the brains of the sham group for both hemispheres. The leptin treatment significantly diminished the number of 'apoptotic cells' in the hippocampal CA1, CA2, CA3, and gyrus dentatus regions in both hemispheres. Leptin treatment significantly preserved the number of neurons in both hemispheres, when compared with the vehicle treated group. CONCLUSION: We conclude that leptin treatment improves neuronal density and decreases apoptosis in the newborn rat with hypoxic-ischemic brain injury. PMID- 21627549 TI - Are we ready for a new look at the diagnosis of premature rupture of membranes? AB - Premature rupture of membranes is a significant contributor to preterm birth with its associated short- and long-term complications. The absence of a standard approach to its management places a burden on the clinicians' ability to promptly and accurately diagnose premature rupture of membranes. For the last half century, there have been no significant changes in the way premature ruptured membranes is diagnosed. With the advent of newer, amniotic fluid-specific, noninvasive, and accurate markers, there is an opportunity to update the diagnosis of premature rupture of membranes. PMID- 21627550 TI - Innate immune system gene polymorphisms in maternal and child genotype and risk of preterm delivery. AB - OBJECTIVE: There is little information about the combination of genetic variability in pregnant women and their children in relation to the risk of preterm delivery (PTD). In a sub-cohort of 487 non-Hispanic white and 288 African American mother/child pairs, the Pregnancy Outcomes and Community Health Study assessed 10 functional polymorphisms in 9 genes involved in innate immune function. METHODS: Race-stratified weighted logistic regression models were used to calculate odds ratios for genotype and PTD/PTD subtypes. Polymorphisms significantly associated with PTD/PTD subtypes were tested for mother/child genotype interactions. RESULTS: Three maternal polymorphisms (IL-1 receptor antagonist intron two repeat (IL-1RN), matrix metalloproteinase- -C1562T, and TNF receptor two M196R (TNFR2)) and three child polymorphisms (IL1-RN, tumor necrosis factor-alpha -G308A, and TNFR2) were associated with PTD, but associations varied by PTD subtype and race. Two interactions were detected for maternal and child genotype. Among non-Hispanic white women, the odds of PTD was higher when both mother and child carried the IL-1RN allele two (additive interaction p < 0.05). Among African-American women, the odds of PTD were higher when both mother and child carried the TNFR2 R allele (multiplicative interaction p < 0.05). CONCLUSION: These results highlight the importance of assessing both maternal and child genotype in relation to PTD risk. PMID- 21627551 TI - The insulin-like growth factor 2 receptor gene Gly1619Arg polymorphism and idiopathic recurrent spontaneous abortion. AB - OBJECTIVE: The insulin-like growth factor 2 (IGF2) and its receptor (IGF2R) are important regulators of placental function and fetal growth. Our aim was to investigate the association between IGF2R gene Gly1619Arg G>A polymorphism and recurrent spontaneous abortion (RSA). METHODS: Polymerase chain reaction and restriction fragment length polymorphism methods were performed to identify the genotypes in 149 couples with a history of at least three consecutive spontaneous abortions and 149 age-matched, unrelated, and fertile couples. RESULTS: There were no statistically significant differences in genotype or allele frequency among the couples with RSA and healthy controls. CONCLUSIONS: We found no evidence that the IGF2R Gly1619Arg variation is associated with RSA. PMID- 21627552 TI - Prenatal diagnosis and cesarean section in a large, population-based birth defects registry. AB - OBJECTIVE: To describe the patterns of cesarean section (CS) and vaginal delivery by type of birth defect and determine whether prenatal diagnosis predicts a higher or lower likelihood of CS for selected defect categories. METHODS: Data from a large population-based registry were analyzed to determine percentages of vaginal versus CS delivery for each of 49 categories of birth defects. Odds ratios and statistical significance were computed to determine if a record of prenatal diagnosis (PND) predicted delivery mode. Cases were liveborn children with any of these defects born in Texas between 1997 and 2005. RESULTS: Forty three percent of infants in the study were delivered by CS, with a range of 25.3% (aniridia) to 62.4% (spina bifida). A record of prenatal diagnosis of the primary assigned birth defect was found in 43.0% of all records but varied substantially by defect category. PND significantly predicted higher CS percentages for spina bifida without anencephaly, encephalocele, hydrocephaly, transposition of the great vessels, ventricular septal defect, pulmonary valve atresia/stenosis, craniosynostosis, diaphragmatic hernia, gastroschisis, and trisomy 21. Vaginal delivery was predicted by PND of anencephaly, agenesis, aplasia, or hypoplasia of the lung, renal agenesis or dysgenesis, and trisomy 18. CONCLUSION: Texas children with birth defects are more likely to have been delivered by CS than the population in general. For several types of defects, prenatal diagnosis is predictive of higher odds of CS; for others, especially fatal defects, PND predicts lower CS likelihood. PMID- 21627553 TI - Serum prohepcidin levels and iron parameters in term small-for-gestational-age newborns. AB - AIM: To understand the effect of prenatal chronic hypoxia on prohepcidin levels in term newborns. METHOD: We determined prohepcidin (Pro-Hep) levels in both term appropriate-for-gestational age (AGA) and term small-for-gestational-age (SGA) infants. Uteroplacental insufficiency had exposed all SGA infants to chronic hypoxia. Serum samples were collected from nine full-term SGA infants. Samples were analyzed for complete blood count, serum iron and ferritin concentrations, iron-binding capacity, and prohepcidin levels. RESULTS: The mean serum Pro-Hep level was 156.4 ? 46.7 ng/ml for SGA infants and 482 ? 371.9 ng/ml for 16 healthy term AGA infants (historical controls); this difference was statistically significant. Statistical analyses revealed significant between-group differences for hemoglobin, hematocrit, mean corpuscular volume, red blood cell distribution width, and serum ferritin and Pro-Hep levels. CONCLUSION: This study showed that compared with AGA infants, Pro-Hep levels were lower in term SGA infants, suggesting that prenatal chronic hypoxia decreases Pro-Hep synthesis. PMID- 21627554 TI - Low maternal vitamin B(12) is a risk factor for neural tube defects: a meta analysis. AB - OBJECTIVE: The objective of this study was to assess whether low level of maternal vitamin B(12) is associated with an increased risk of fetal neural tube defects (NTDs), in order to contribute to research on further reduction of NTDs under a background of mandatory folic acid (FA) fortification. METHODS: A meta analysis was conducted. We retrieved and evaluated the studies published on the risk of low level of maternal vitamin B(12) for NTDs. The homogeneity of the studies was examined using the forest graph. Meta-analysis was applied to calculate the odds ratio (OR) of fetal NTDs in relation to low maternal B(12) and its 95% confidence interval (CI). RESULTS: We identified nine published articles including 567 cases and 1566 controls in the meta-analysis. All the studies selected were homogeneous according to the forest graph (chi(2) = 15.05, P < 0.1). The estimated OR value of fetal NTDs in relation to low maternal B(12) was 2.41 (95% CI: 1.90-3.06). CONCLUSION: Low maternal B(12) status could be an important risk factor for the development of fetal NTDs. The addition of synthetic B(12) to current recommendations for periconceptional FA tablet supplements or FA-fortified foods should be considered. PMID- 21627555 TI - State of the art in hard-on-hard bearings: how did we get here and what have we achieved? AB - Total hip arthroplasty has shown excellent results in decreasing pain and improving function in patients with degenerative disease of the hip. Improvements in prosthetic materials, designs and implant fixation have now resulted in wear of the bearing surface being the limitation of this technology, and a number of hard-on-hard couples have been introduced to address this concern. The purpose of this article is to review the origins, development, survival rates and potential advantages and disadvantages of the following hard-on-hard bearings for total hip arthroplasty: metal-on-metal standard total hip arthroplasty; metal-on-metal hip resurfacing arthroplasty, ceramic-on-ceramic total hip arthroplasty; and ceramic on-metal bearings. Improvements in the manufacturing of metal-on-metal bearings over the past 50 years have resulted in implants that provide low wear rates and allow for the use of large femoral heads. However, concerns remain regarding elevated serum metal ion levels, potential teratogenic effects and potentially devastating adverse local tissue reactions, whose incidence and pathogenesis remains unclear. Modern total hip resurfacing has shown excellent outcomes over 10 years in the hands of experienced surgeons. Current ceramic-on-ceramic bearings have demonstrated excellent survival with exceptionally low wear rates and virtually no local adverse effects. Concerns remain for insertional chipping, in vivo fracture and the variable incidence of squeaking. Contemporary ceramic-on metal interfaces are in the early stages of clinical use, with little data reported to date. Hard-on-hard bearings for total hip arthroplasty have improved dramatically over the past 50 years. As bearing designs continue to improve with new and modified materials and improved manufacturing techniques, it is likely that the use of hard-on-hard bearings will continue to increase, especially in young and active patients. PMID- 21627556 TI - Symmetry in number of retrieved oocytes between two ovaries: a possible predictor of in vitro fertilization outcome. AB - OBJECTIVE: To examine the relationship between the ratio of difference in number of retrieved oocytes from the two ovaries to total number of oocytes (difference ratio, |N(RO) - N(LO)|/N(TOTAL)) and in?vitro fertilization (IVF) outcome. METHODS: Retrospective review of medical record for infertility patients with intact two ovaries who had undertaken controlled ovarian stimulation (COS) and IVF cycles in Seoul National University Hospital was conducted. Characteristics and IVF outcomes were compared according to the difference ratio (|N(RO) - N(LO)|/N(TOTAL)) using student t-test and chi2 test. To adjust for the effect of confounding factors, binary logistic regression test was performed. RESULTS: When the study population was divided according to the difference ratio by cut-off value of 0.4, patients with difference ratio less than 0.4 showed significantly higher pregnancy rate (25.7% vs. 17.1%, p = 0.021). A significant correlation was noted between the difference ratio and achievement of pregnancy after adjustment for patient's age and total number of top quality embryos transferred (adjusted OR = 0.40, 95% CI 0.19-0.83, p = 0.014). CONCLUSIONS: The difference ratio of retrieved oocytes in patients with intact two ovaries was observed to have prognostic value in IVF cycles. PMID- 21627557 TI - Serum anti-Mullerian hormone, inhibin B, and total inhibin levels in women with hypothalamic amenorrhea and anorexia nervosa. AB - OBJECTIVE: To evaluate whether neuroendocrine forms of secondary amenorrhea (hypothalamic nervosa (HA) and anorexia nervosa (AN)) affect serum anti-Mullerian hormone (AMH), inhibin B, and total inhibin levels. METHODS: Amenorrheic women (n = 82) (aged between 16 and 35 years old) according to diagnosed with neuroendocrine forms of amenorrhea: HA (n = 64), AN (n = 18), and healthy women (n = 41) (control group) were enrolled. Serum AMH, inhibin B, and total inhibin levels were measured by specific ELISA. RESULTS: No statistically significant difference of AMH serum levels between women with HA, AN, and control group was observed. Serum inhibin B and total inhibin levels in women with HA (p < 0.0001), AN (p < 0.05) resulted significantly lower than in control healthy women. CONCLUSION: The present data showed that neuroendocrine forms of amenorrhea are associated with an impaired inhibin secretion while not AMH. These data indirectly support that AMH is an excellent marker of ovarian reserve and its secretion is not influenced by the hypothalamic-ovarian axis activity. PMID- 21627558 TI - The effect of risedronate treatment on serum osteoprotegerin and bone marker levels in postmenopausal women with osteoporosis. AB - BACKGROUND: To evaluate the effect of risedronate treatment on osteoprotegerin (OPG), C-terminal cross-linking telopeptide of type 1 collagen (CTX), osteocalcin (OC), and deoxypyridinoline (DPD). METHODS: Eighty postmenopausal osteoporotic patients were randomized into two groups. In first group, patients received 35?mg of risedronate once a week and calcium with vitamin D per day. In second group, patients received only calcium with vitamin D per day. Bone turnover markers were measured at baseline, 1st, 3rd and 6th month. RESULTS: OPG levels were significantly reduced at 1st and 6th month of treatment in both groups, but no statistically significant difference was detected between groups. In the group treated with risedronate, difference in CTX level was observed at 3rd month of treatment, while a difference in DPD and OC levels were observed at 6th month of treatment. The baseline OPG levels correlated with age, menopause duration, and CTX levels. There was no correlation between OPG levels and the levels of the other markers during treatment. CONCLUSION: The present study showed that using risedronate in treatment of postmenopausal osteoporosis causes no specific changes in OPG levels; therefore, in contrast to some of the studies in the literature OPG may not be useful marker in monitoring of bisphosphonate. PMID- 21627559 TI - Spinal multifocal amyloidosis derived from wild-type transthyretin. AB - Abstract Spinal amyloidosis can occur as a part of systemic amyloidosis or as localized amyloidomas. However, the exact pathogenesis of the spinal amyloidosis remains to be fully understood. Transthyretin (TTR) is an amyloidogenic protein causing two kinds of amyloid diseases. One is senile systemic amyloidosis (SSA), which is caused by wild-type (WT) TTR and primarily affects cardiac functions. The other type is familial amyloidosis, which is mainly induced by mutated TTR. We report here the first case of multifocal spinal TTR amyloidosis derived from WT TTR with radiculomyelopathy and destructive spondylosis. The data and clinical manifestations suggest that the patient may develop SSA. Clinical manifestations of TTR-related amyloidosis may vary more than we previously thought. In spinal amyloidosis, WT TTR is one of the candidate precursor proteins for the disease. PMID- 21627560 TI - Ageing: a risk factor for amyloid A amyloidosis in rheumatoid arthritis. AB - OBJECTIVE: Acceleration of amyloid A (AA) amyloidosis induction by ageing has not been extensively studied in rheumatoid arthritis (RA). The aim of this study is to clarify contribution of ageing to the development of AA amyloidosis associated with RA in our large cohort. METHODS: 388 adult-onset RA patients whose RA was complicated by biopsy-proven AA amyloidosis were enrolled. The ages of RA onset and AA amyloidosis diagnosis were estimated in each patient. The contributions of ageing, inflammatory activity, SAA1 exon 3 polymorphism as well as gender to the pathogenesis of AA amyloidosis in 144 cases were also studied by multiple regression analysis. RESULTS: Subjects with RA onset at older age had a shorter period to develop amyloidosis than those with disease onset at younger age (p < 0.001). The interval between RA onset and AA amyloidosis diagnosis was significantly shorter in the SAA1.3 positive group than in the SAA1.3 negative (p=0.001). Multiple regression analysis indicated that the interval from RA onset to diagnosis of AA amyloidosis is determined by age at RA onset (p < 0.001), the most recent median annual CRP concentration (p=0.006) and SAA1.3 allele (p=0.058). Gender did not significantly contribute to the onset of AA amyloidosis (p=0.569). CONCLUSION: Ageing is an independent risk factor for the induction of AA amyloidosis complicating RA. PMID- 21627561 TI - Circadian rhythm of hot flashes and activity levels among prostate cancer patients on androgen deprivation therapy. AB - OBJECTIVE: Altered circadian rhythms have been identified in untreated prostate cancer patients. Findings of restored rhythmicity following cancer treatment may have relevance for cancer control and symptom management. This study assessed and compared the cyclic patterns of hot flashes and activity levels in treated prostate cancer patients. METHODS: Data were collected during two 24-h periods among 47 prostate patients undergoing androgen deprivation therapy (ADT). Hot flashes were detected objectively through sternal skin conductance and by patients via electronic event marking. Activity levels were recorded on a wrist actigraphy device. RESULTS: The mean frequency of objectively measured and patient-reported hot flashes was 13.6 (SD = 14.3) and 12.6 (SD = 9.6), respectively. There were significant 24-h circadian rhythms of both hot flashes and activity levels. The peak of the rhythms occurred in early afternoon. There was no significant cross correlation between hot flashes and activity levels. CONCLUSIONS: The acrophases of hot flashes and elevated activity levels in this study may represent a normalisation of circadian rhythms following ADT, pointing to the need for more research, including controlled, prospective chronobiologic studies. Future research may have important implications for the survival of prostate cancer patients and the identification of new and safe hot flash treatments. PMID- 21627562 TI - Change in testosterone concentrations over time is a better predictor than the actual concentrations for symptoms of late onset hypogonadism. AB - BACKGROUND: Symptoms of late-onset hypogonadism (LOH) and concentrations of testosterone (T) and bioavailable testosterone (BT) were studied in relation to the data from the same men 5 years earlier. METHODS: In 2008, 282 men, aged 60-82 years, answered a questionnaire regarding demographic data, medical history, different symptoms of LOH and the 10 questions from the 'Androgen Decline in Aging Males (ADAM)-questionnaire'. Blood samples were analysed for concentrations of T and calculations were made for BT. RESULTS: A total of 87.2% of the questionnaires were returned and analysed, and 75.2% of the responders gave blood samples. The oldest third of the men were most affected by LOH symptoms (p < 0.05). Both T and BT concentrations decreased during the 5 years (p < 0.05) but only the symptom 'less strong erections' changed significantly (p < 0.05). Men reporting one of the four specific symptoms from the 'ADAM-questionnaire' for the first time in 2008 had a higher loss of T and BT than men who had unchanged or fewer symptoms than that reported in 2003. CONCLUSIONS: The magnitude of the decrease in concentrations is a better predictor of LOH than are the actual concentrations of T and BT. A combination of symptoms predicts LOH better than any single symptom. PMID- 21627563 TI - Radioresistance development of DNA repair deficient Escherichia coli DH5alpha in ground beef subjected to electron beam at sub-lethal doses. AB - PURPOSE: Electron beam (e-beam) efficiently and non-thermally inactivates microorganisms in food by lethal DNA changes (direct effects) and free radicals from water radiolysis (in-direct effects). Non-pathogenic Escherichia coli DH5alpha (alpha substrain of DH5 described by Hanahan 1985 , 'DH' stands for Douglas Hanahan) is a microorganism that lacks DNA repair capability, resulting in high radiosensitivity. Studying microbial inactivation of E. coli DH5alpha repeatedly subjected to sub-lethal e-beam in ground beef may enhance understanding of microbial radioresistance. The objective of this study was to determine if repetitive processing with e-beam at sub-lethal doses increases D value (e-beam dose required to inactivate one log of microbial population) of E. coli DH5alpha in ground beef. MATERIALS AND METHODS: Survivors from the highest e beam dose were isolated and incubated in ground beef for the next cycle of e-beam processing. Five cycles were conducted. To acclimatise E. coli DH5alpha, first two cycles used low doses. D-values were determined following the third cycle. RESULTS: D-values increased (p < 0.05) significantly with each cycle. Thus, E. coli DH5alpha has a capability to develop greater radioresistance under these experimental conditions. Following the third cycle D-values were 0.32 +/- 0.006 and 0.32 +/- 0.002 kGy for survivors enumerated on non-selective and selective media, respectively; the fourth cycle 0.39 +/- 0.007 and 0.40 +/- 0.019 kGy; and the fifth cycle 0.46 +/- 0.006 and 0.46 +/- 0.020 kGy. D-values on non-selective and selective media were similar (p > 0.05) indicating absence of cell recovery in E. coli DH5alpha. CONCLUSIONS: E. coli DH5alpha increases radioresistance to e beam as a result of repetitive exposure to sub-lethal doses despite its DNA repair deficiency. PMID- 21627564 TI - Topical application of hPDGF-A-modified porcine BMSC and keratinocytes loaded on acellular HAM promotes the healing of combined radiation-wound skin injury in minipigs. AB - PURPOSE: To evaluate the efficacy of cultured cutaneous substitute (CCS) in accelerating the healing of combined radiation-skin wound injury (CRWI) in minipigs. MATERIAL AND METHODS: Autologous porcine bone marrow-derived mesenchymal stem cells (BMSC) and skin-derived keratinocytes (SK) were infected by recombinant retrovirus expressing human (h) platelet-derived growth factor-A (hPDGF-A). CCS was constructed by loading acellular human amniotic membrane (HAM) with normal porcine BMSC and SK (BMSC-/SK-CCS) or with hPDGF-A modified counterparts (BMSC+/SK+CCS). The expression of exogenous hPDGF-A in cells and CCS was assessed by reverse transcription polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA). The CCS or HAM were grafted to the dorsal CRWI sites (20 Gy local irradiation plus full-thickness skin removal, diameter = 40 mm) of minipigs. Wound healing rate and pathological changes were observed. RESULTS: High levels of hPDGF-A expression were confirmed in gene modified cells (3780 pg/ml), cultured CCS (506 pg/ml) and transplanted CCS (250 pg/ml). The transplantation of the BMSC+/SK+CCS resulted in a shorter healing time (16-18, days) (P < 0.05 vs. other groups). The healing rates ranked as BMSC+/SK+CCS > BMSC-/SK-CCS > HAM > wound control. Pathologically, there were better granulation formation and re-epithelialisation, and collagen deposition in BMSC+/SK+CCS-treated wound than those in other groups. The angiogenesis ability followed the same order as healing rate of different groups. At day 7, the area densities of vasculature in granulation tissue of group BMSC+/SK+CCS, BMSC-/SK CCS, HAM, wound only were 15.4, 10.3, 6.0 and 5.7%, respectively, while the number densities of vasculature was 767, 691, 126 and 109 (number/mm(2)), respectively. CONCLUSIONS: Topical transplantation of hPDGF-A modified CCS may be applicable to the management of refractory wounds. PMID- 21627565 TI - hTERT regulation by NF-kappaB and c-myc in irradiated HER2-positive breast cancer cells. AB - PURPOSE: Telomerase activity (TA), frequently observed in cancer, compensates for telomere shortening thus preventing cell senescence and conferring resistance to therapy. In the present study, we investigated the expression of human telomerase reverse transcriptase (hTERT) and TA and their regulation, as well as apoptotic rates and correlation with the presence of human epidermal growth factor receptor 2 (HER2), in irradiated tumour-derived breast cancer cells. MATERIALS AND METHODS: In 50 breast cancer tissue samples hTERT mRNA expression and TA were correlated with cell features (HER2, Estrogen and Progesterone Receptor status). Cells from six samples were then irradiated with 10 and 20 Gy; apoptotic rates were measured by flow cytometry, hTERT mRNA expression by real-time polymerase chain reaction and TA by telomeric repeat amplification protocol assay, at 24-144 h post-irradiation. Chromatin immunoprecipitation was performed to investigate hTERT and cellular-myelocytomatosis (c-myc) promoters' activity. HER2 gene knockdown was performed using small interfering RNA technology. RESULTS: hTERT/TA were found increased only in irradiated HER2-positive cells, which were found to be more radioresistant, while HER2 knockdown led to hTERT/TA downregulation. HER2 was found to mediate hTERT expression through activation of Nuclear Factor-kappa B (NF-kappaB) and c-myc. CONCLUSIONS: The present study suggests that following irradiation, HER2 receptor activates hTERT/telomerase, increasing the breast cancer cells' survival potential, through sequential induction of transcription factors NF-kappaBeta and c-myc. PMID- 21627566 TI - Effect of dose-rate on induction of neoplastic transformation in vitro by low doses of 232 MeV protons. AB - PURPOSE: To determine the effect of dose-rate on induction of neoplastic transformation in vitro by low doses of 232 MeV protons. MATERIALS AND METHODS: The experimental system used was the human hybrid cell assay. The dose-rates examined were 50 cGy/min and 20 cGy/h. The dose-rate 20 cGy/h was chosen as this is in the range of the maximum dose-rate that can be experienced in an unshielded space environment following a solar flare. At low dose-rate (LDR), doses from 0.5 100 cGy were studied. At high dose rate (HDR), the dose range was 0.5-200 cGy. RESULTS: The data indicated no significant differences between the two dose-rates at doses up to 100 cGy. CONCLUSION: For the endpoint of neoplastic transformation in vitro, high dose-rate data may be sufficient to estimate low dose-rate effects (20 cGy/h) in the dose range up to 100 cGy from 232 MeV protons. The data are of relevance to risk estimation for space travel. PMID- 21627567 TI - Genetic analysis of HIV type 1 env gene in cerebrospinal fluid and plasma of infected Chinese paid blood donors. AB - HIV infection in the central nervous system (CNS) can progress to AIDS dementia complex (ADC). Currently, the HIV-1 env gene in the CNS of infected Chinese paid blood donors (PBDs) has not been well characterized. In the study, the C2-V5 regions of the HIV env gene were cloned and sequenced from both cerebrospinal fluid (CSF) and plasma samples of six HIV-infected Chinese PBDs. Sequence analysis revealed that the sequences from Henan province clustered closely with subtypes B' and B, and the levels of diversity from the CNS were significantly lower than those from blood (p<0.0001). In addition, all viral quasispecies from CNS use CCR5 as the coreceptor. These data provide valuable information on HIV pathogenesis in the CSF and plasma of infected Chinese PBDs, and our findings could enhance insights into HIV-associated neurological disease. PMID- 21627568 TI - Effect of in vitro passaging on the stem cell-related properties of tendon derived stem cells-implications in tissue engineering. AB - This study aimed to compare clonogenicity, proliferation, stem cell-related marker expression, senescence, and differentiation potential of rat patellar tendon-derived stem cells (TDSCs) at early (P5), mid (P10), and late (P20, P30) passages. The clonogenicity of the cells was assessed by colony-forming assay and their proliferative potential was assessed by bromodeoxyuridine assay. The surface expression of CD90 and CD73 was assessed by flow cytometry. The cellular senescence was assessed by beta-galactosidase activity. The adipogenic, chondrogenic, and osteogenic differentiation potentials of TDSCs were assessed by standard assays after induction. The mRNA expression of tendon-related markers, scleraxis (Scx) and tenomodulin (Tnmd), was measured by quantitative real-time reverse transcription-polymerase chain reaction. Both the colony numbers and proliferative potential of TDSCs increased with passaging. Concomitantly, there was significant upregulation of beta-galactosidase activity with TDSC passaging. The subculture of TDSCs downregulated the expression of CD90 and CD73. Lipid droplets were formed in the early and mid passages of TDSCs upon adipogenic induction, but were absent in the late passages. The expression of peroxisome proliferator activator receptor gamma 2 (PPARgamma2) and CCAAT/enhancer binding protein alpha (C/EBPalpha) in TDSCs after adipogenic induction decreased with passaging. Chondrogenesis, proteoglycan deposition, collagen type II protein expression, collagen type 2A1 (Col2AI), and aggrecan (Acan) mRNA expression were less in pellets formed with later passages of TDSCs after chondrogenic induction. The expression of Scx and Tnmd was lower in the late, compared with early and mid, passages of TDSCs. However, matrix mineralization and expression of alkaline phosphatase (Alpl) and osteocalcin (Bglap) mRNA after osteogenic induction increased with TDSC passaging. Researchers and clinicians should consider the changes of stem cell-related properties of TDSCs when multiplying them in vitro for tissue engineering. PMID- 21627569 TI - Induction of cardiomyogenesis in human embryonic stem cells by human embryonic stem cell-derived definitive endoderm. AB - We previously reported that chick anterolateral endoderm (AL endoderm) induces cardiomyogenesis in mouse embryoid bodies. However, the requirement to micro dissect AL endoderm from gastrulation-stage embryos precludes its use to identify novel cardiomyogenic factors, or to scale up cardiomyocyte numbers for therapeutic experiments. To circumvent this problem we have addressed whether human definitive endoderm (hDE) cells, which can be efficiently generated in large numbers from human embryonic stem cells (hESCs), can mimic the ability of AL endoderm to induce cardiac myogenesis. Results demonstrate that both hDE cells and medium conditioned by them induce cardiac myogenesis in pluripotent hESCs, as indicated by rhythmic beating and immunohistochemical/quantitative polymerase chain reaction monitoring of marker gene expression. The cardiomyogenic effect of hDE is enhanced when pluripotent hESCs are preinduced to the mes-endoderm state. Because this approach is tractable and scalable, it may facilitate identification of novel hDE-secreted factors for inclusion in defined cardiomyogenic cocktails. PMID- 21627570 TI - Plasma antioxidants are associated with impaired lung function and COPD exacerbations in smokers. AB - Low molecule weight antioxidants such as uric acid (UA), glutathione (GSH), and ascorbate (ASC) counter the effects of oxidants produced by cigarette smoke. Although dietary intake of foods rich in antioxidants has been associated with a reduced risk of smokers developing chronic obstructive pulmonary disease (COPD), the association between plasma antioxidants and COPD is less clear. In this cross sectional study we investigated the relationship among plasma antioxidants and COPD phenotypes (severity of airflow obstruction on spirometry and history of exacerbations) in 136 smokers with normal lung function and 367 smokers with COPD. In the multivariate analysis, a lower plasma UA was associated with more severe COPD (P < 0.002) and a lower GSH was associated with a history of COPD exacerbations (P = 0.03); ASC was not associated with any COPD phenotypes. This suggests that antioxidant balance is impaired in smokers with obstruction on spirometry or a history of COPD exacerbations. PMID- 21627571 TI - Why we must "TRANSFUSE". PMID- 21627572 TI - CHEST and the impact of fraud in fluid resuscitation research. PMID- 21627573 TI - Critical Care and Resuscitation's response to the Boldt scandal. PMID- 21627574 TI - Calcium flux in continuous venovenous haemodiafiltration with heparin and citrate anticoagulation. AB - BACKGROUND: Calcium chelation with citrate is an effective alternative to heparin for anticoagulation of the extracorporeal circuit during continuous venovenous haemodiafiltration (CVVHD-F). Calcium release occurs upon citrate metabolism; however, ultrafiltration of citrate-bound and free ions also occurs. OBJECTIVE: To quantify calcium loss and improve understanding of calcium homeostasis in CVVHD-F. METHODS: Calcium loss was prospectively quantified from heparinised and citrated circuits in consecutive intensive care patients requiring CVVHD-F. CVVHD F prescription and anticoagulation choice was by the treating intensivist using commercial solutions (Gambro, Lundia, Sweden). Sample sets comprising arterial, prefilter and postfilter blood and an effluent sample were analysed for ionised total calcium (iCa(2+)) and magnesium levels. Flow rates were then used to calculate calcium flux. Citrate dose (predilution rate) and calcium replacement followed unit protocols to maintain a circuit iCa(2+) concentration of 0.3-0.5 mmol/L and an arterial iCa(2+) concentration of 0.8-1.1 mmol/L. RESULTS: 26 heparinised circuits and 22 citrated circuits in 13 patients were included; 334 sample sets were tested. For target extracorporeal blood flows of 200 mL/min, mean predilution Prismocitrate 10/2 flows were 1660 mL/h, delivering 2.42 mmol citrate per litre of blood. For heparin, mean predilution flows of Hemosol B0 were 2058mL/h. Mean calcium loss was 4.01 mmol/h from citrate anticoagulated circuits versus a gain of 0.24mmol/h from heparinised circuits (P < 0.001). Despite calcium replacement, citrate patients experienced a mean calcium loss of 1.12 mmol/h (SD, 0.70; 95% CI 1.0-1.22mmol/h; P < 0.001). Calculated effective diffusion volume (Q(E)) for calcium was closer to total blood water volume in heparin circuits and closer to plasma water volume in citrate circuits. CONCLUSIONS: Despite supplementation to maintain arterial iCa(2+) levels, citrate anticoagulation results in a net calcium deficit. An equation for estimating required citrate dose may allow revision of citrate dosing protocols. PMID- 21627575 TI - Bedside electronic capture of clinical observations and automated clinical alerts to improve compliance with an Early Warning Score protocol. AB - BACKGROUND: Failure to comply with clinical protocols and failure of communication to ensure delivery of the most appropriate timely clinical responses to patients whose conditions are acutely deteriorating have been shown to be significant causative factors associated with inhospital adverse events. OBJECTIVE: To determine whether automated clinical alerts increase compliance with an Early Warning Score (EWS) protocol and improve patient outcomes. METHODS: We performed a historically controlled study of bedside electronic capture of observations and automated clinical alerts. The primary outcome measure was hospital length of stay (LOS); secondary outcome measures were compliance with the EWS protocol, cardiac arrest incidence, critical care utilisation and hospital mortality. RESULTS: Between baseline and intervention, 1481 consecutive patients were recruited generating 13 668 observation sets. There was a reduction in hospital LOS between the baseline and alert phase (9.7 days v 6.9 days, P < 0.001). EWS accuracy improved from 81% to 100% with electronic calculation. Clinical attendance to patients with EWS 3, 4 or 5 increased from 29% at baseline to 78% with automated alerts (P < 0.001). For patients with an EWS > 5, clinical attendance increased from 67% at baseline to 96% with automatic alerts (P < 0.001). CONCLUSIONS: Electronic recording of patient observations linked to a computer system that calculates patient risk and then issues automatic graded alerts can improve clinical attendance to unstable general medical ward patients. PMID- 21627576 TI - Analysis of orotracheal intubation techniques in the intensive care unit. AB - BACKGROUND: The development of specialised airway tools help laryngoscopists secure the airway in intensive care units. The use of videolaryngoscopy has been suggested in simulation studies, and human studies suggest that this advanced airway tool may have an advantage for difficult airways; however, less is known about its use in the ICU. OBJECTIVE: To compare orotracheal intubation before and after acquisition of an ICU-dedicated GlideScope (GS), and to determine the incidence of complications with orotracheal intubation in an ICU. METHODS: An observational study was conducted from October 2008 to April 2009 to record the use of advanced airway tools including videolaryngoscopy before ("pre-GS") and immediately after ("post-GS") the purchase of an ICUdedicated videolaryngoscope. Reasons for intubation, response time, type of intubation, number of attempts at intubation, reasons for delays in intubation, risk factors for difficult intubation and complications were compared between these groups. RESULTS: 56 patients were intubated pre-GS and 47 post- GS. Although a significant increase in videolaryngoscopy was observed in the ICU (P = 0.001), no significant reduction in total attempts at orotracheal intubation were observed (P = 0.66), and that the incidence of overall complications were not reduced (P = 0.21). CONCLUSIONS: The use of a new airway tool may not necessarily lead to immediate reduction in attempts at orotracheal intubation or in overall complication rates. PMID- 21627577 TI - Fever and fever management among intensive care patients with known or suspected infection: a multicentre prospective cohort study. AB - OBJECTIVE: To describe the duration of fever, fever management, and outcomes among intensive care patients with fever and known or suspected infection. DESIGN, SETTING AND PARTICIPANTS: Prospective observational trial in three tertiary intensive care units over 6 weeks in 2010. Adult patients were screened for eligibility and inclusion if they had a fever of >=38.0 degrees C and known or suspected infection being treated with antimicrobials; those with neurological injury or elective surgery within 72 hours were excluded. MAIN OUTCOME MEASURES: Mean and peak daily temperatures were recorded and the use of antipyretics and other cooling measures were recorded over the first 7 days. Mortality, ICU-free survival, ventilator-free survival and renal replacement therapy-free survival were determined at Day 28. RESULTS: 51/565 patients (9.0%) were included. The mean daily peak temperature and the proportion of patients with a documented temperature of >=38.0 degrees C decreased over the first 3 days after first documented fever. Thereafter, the proportion of patients who had daily peak temperatures >=38.0 degrees C remained about 20%. Paracetamol was administered to 58%-70% of patients per day. Physical cooling was used at least once for 12% of patients. Mean ICU-free survival to Day 28 in eligible patients was 16.0 (SD, 9.2) days. The mortality rate of eligible patients was more than double that of ineligible patients (8/51 [16%] v 36/514 [7%]; P = 0.05). CONCLUSION: We have described the typical time course of fever in an easily identified cohort of patients with known or suspected infection and have determined that these patients have significant morbidity and mortality. This information is vital to the design of interventional studies for the treatment of fever in ICU. PMID- 21627578 TI - Epidemiology and significance of coagulase-negative staphylococci isolated in blood cultures from critically ill adult patients. AB - BACKGROUND: Little published data are available on the epidemiology and significance of coagulase-negative staphylococci (CoNS) in blood culture isolates among critically ill adult patients. OBJECTIVES: To describe the epidemiology and frequency of CoNS blood culture isolates in critically ill adults, and investigate the association between time to positivity (TTP) of blood cultures and number of culture-positive bottles with organ dysfunction and mortality. DESIGN, SETTING AND PARTICIPANTS: A retrospective chart audit in the intensive care unit of a tertiary hospital comprising all patients who had positive blood cultures for CoNS in 2009. MAIN OUTCOME MEASURES: TTP, number of culture-positive bottles, Sequential Organ Failure Assessment (SOFA) scores, resolution of fever and white cell response and inotrope requirement, length of stay in ICU and mortality. RESULTS: In 2009, there were 1514 and 109 positive blood culture sets for the hospital and ICU patients, respectively. Of these, 515 sets from patients outside the ICU (34% of all hospital positive blood cultures) and 54 from the ICU (49.5% of all ICU positive blood cultures) were positive for CoNS. Patients with TTP <=24 hours had higher organ failure scores by 0.9 (95% CI, 0-3.4; P = 0.052). There was a trend towards an association between increased 28-day mortality and TTP <=24 hours (7/22 v 3/32; P = 0.071). There was no significant correlation between number of bottles positive for culture and mortality, length of stay, SOFA score, resolution of fever, white cell response, and inotrope requirement. CONCLUSIONS: Early TTP of blood cultures with CoNS may be associated with poorer outcome and may be a marker of true infection. Given the relatively high frequency of this microbiological problem, larger prospective observational studies are required to more clearly define the significance of a CoNS blood culture isolates in critically ill adult patients. PMID- 21627579 TI - Increased blood volume following resolution of acute cardiogenic pulmonary oedema: a retrospective analysis. AB - BACKGROUND: Acute cardiogenic pulmonary oedema (APO) occurs due to an increase in pulmonary microvascular pressure and massive transvascular fluid filtration into the lungs, causing respiratory insufficiency. OBJECTIVE: To determine whether fluid sequestration in the lungs effectively leads to contraction of the circulating blood volume, leading to relative hypovolaemia, and whether resolution of APO and fluid shift to the vascular compartment restores the circulating volume. METHODS: A retrospective analysis was conducted in the intensive care unit of a university teaching hospital, April - September 2007. It comprised a cohort of APO patients and a control group of patients with acute exacerbation of chronic obstructive pulmonary disease (COPD) with similar demographics. Patient demographics, haematocrit, haemoglobin levels, total protein and albumin levels, and arterial blood gas were analysed at presentation and after clinical resolution or at 24 hours. Fluid balance charts were reviewed. Blood, plasma and cell volume changes were calculated using haemoglobin levels and haematocrit. RESULTS: 52 patients (27 with APO; 25 with COPD) were included. Median haematocrit decreased significantly and the calculated blood and plasma volumes showed statistically significant increases after treatment in the APO group when compared with the COPD group (P < 0.001). Fluid intake and output were well balanced in both groups. CONCLUSIONS: Patients with APO are hypovolaemic at the onset relative to their state after treatment. With treatment and resolution of APO, hypovolaemia is corrected and circulating volume is restored. PMID- 21627580 TI - Liver transplantation rapidly stops cerebral ammonia uptake in fulminant hepatic failure. AB - This report describes the effect of liver transplantation on transcerebral ammonia uptake in a case of fulminant hepatic failure. A young woman with fulminant hepatic failure and coma received monitoring of transcerebral ammonia uptake before and after orthotopic liver transplantation. Before liver transplantation, median transcerebral ammonia uptake was 8 umol/L. After liver transplantation, ammonia uptake decreased to 0 umol/L. Fulminant hepatic failure is associated with transcerebral ammonia uptake, which is fully and rapidly corrected by liver transplantation. PMID- 21627581 TI - Comparison of intermittent haemodialysis, prolonged intermittent renal replacement therapy and continuous renal replacement haemofiltration for lithium toxicity: a case report. AB - This case report compares three types of renal replacement therapy for acute severe lithium toxicity and is the first to use prolonged intermittent renal replacement therapy (PIRRT). A peak lithium level of 13.2mmol/L was recorded after a 51- year-old man attempted suicide. He was treated with intermittent haemodialysis (IHD), PIRRT and continuous venovenous haemofiltration (CVVH) over 55 hours. Lithium clearance was shown to be comparable between IHD and PIRRT, both of which were superior to CVVH. Expected efficacy of treatment with a single daily session of PIRRT was higher than IHD or CVVH. PIRRT is a useful strategy for dialysis among patients with acute lithium intoxication. PMID- 21627582 TI - Bilateral adrenal haemorrhage in a critically ill patient. AB - Although rare, bilateral adrenal haemorrhage remains a life-threatening complication of severe infection and prolonged critical illness. A 67-year-old woman developed acute adrenal haemorrhage in the context of severe systemic infection due to diverticulitis and pericolic abscess. The prompt recognition and management of this condition was an important component of her eventual recovery. PMID- 21627583 TI - The effect of antipyretic medications on mortality in critically ill patients with infection: a systematic review and meta-analysis. AB - BACKGROUND: Antipyretic medications are widely used in critically ill patients with infection despite evidence supporting a protective, adaptive role of fever. OBJECTIVE: To assess the mortality risk of antipyretic medications among critically ill patients with infection. METHODS: A systematic search of MEDLINE, Embase, the Cochrane Central Register of Controlled Trials and PubMed was undertaken to identify randomised controlled trials (RCTs) of antipyretic use among critically ill patients with suspected or confirmed infection that reported mortality. A quantitative meta-analysis of the risk of death was carried out with calculation of the pooled risk of death and standard evaluation of heterogeneity. RESULTS: Six RCTs investigating the use of paracetamol (1) and non-steroidal anti inflammatory medications (5) met the inclusion criteria for meta-analysis. The trials were heterogeneous in terms of study populations and interventions, were not primarily designed to evaluate antipyretic effect on mortality risk, and significant confounding was present from the use of other concomitant antipyretic strategies. The pooled estimates of odds ratios for mortality with antipyretic treatment were 0.96 (95% CI, 0.68-1.34) and 1.08 (95% CI, 0.60-1.96) for fixed effects and random effects, respectively, and the I-squared value was 34.9 (95% CI, 0.0-73.9). CONCLUSION: The studies included in this review were insufficient to allow a robust estimate of the effect of pharmacological antipyresis on mortality in critically ill patients with suspected infection. Further RCTs are required to resolve this important area of clinical uncertainty. PMID- 21627584 TI - Variation in the organization and subunit composition of the mammalian pyruvate dehydrogenase complex E2/E3BP core assembly. AB - Crucial to glucose homoeostasis in humans, the hPDC (human pyruvate dehydrogenase complex) is a massive molecular machine comprising multiple copies of three distinct enzymes (E1-E3) and an accessory subunit, E3BP (E3-binding protein). Its icosahedral E2/E3BP 60-meric 'core' provides the central structural and mechanistic framework ensuring favourable E1 and E3 positioning and enzyme co operativity. Current core models indicate either a 48E2+12E3BP or a 40E2+20E3BP subunit composition. In the present study, we demonstrate clear differences in subunit content and organization between the recombinant hPDC core (rhPDC; 40E2+20E3BP), generated under defined conditions where E3BP is produced in excess, and its native bovine (48E2+12E3BP) counterpart. The results of the present study provide a rational basis for resolving apparent differences between previous models, both obtained using rhE2/E3BP core assemblies where no account was taken of relative E2 and E3BP expression levels. Mathematical modelling predicts that an 'average' 48E2+12E3BP core arrangement allows maximum flexibility in assembly, while providing the appropriate balance of bound E1 and E3 enzymes for optimal catalytic efficiency and regulatory fine-tuning. We also show that the rhE2/E3BP and bovine E2/E3BP cores bind E3s with a 2:1 stoichiometry, and propose that mammalian PDC comprises a heterogeneous population of assemblies incorporating a network of E3 (and possibly E1) cross bridges above the core surface. PMID- 21627585 TI - Palmitoylation of human proteinase-activated receptor-2 differentially regulates receptor-triggered ERK1/2 activation, calcium signalling and endocytosis. AB - hPAR(2) (human proteinase-activated receptor-2) is a member of the novel family of proteolytically activated GPCRs (G-protein-coupled receptors) termed PARs (proteinase-activated receptors). Previous pharmacological studies have found that activation of hPAR(2) by mast cell tryptase can be regulated by receptor N terminal glycosylation. In order to elucidate other post-translational modifications of hPAR(2) that can regulate function, we have explored the functional role of the intracellular cysteine residue Cys(361). We have demonstrated, using autoradiography, that Cys(361) is the primary palmitoylation site of hPAR(2). The hPAR(2)C361A mutant cell line displayed greater cell-surface expression compared with the wt (wild-type)-hPAR(2)-expressing cell line. hPAR(2)C361A also showed a decreased sensitivity and efficacy (intracellular calcium signalling) towards both trypsin and SLIGKV. In stark contrast, hPAR(2)C361A triggered greater and more prolonged ERK (extracellular-signal regulated kinase) phosphorylation compared with that of wt-hPAR(2) possibly through Gi, since pertussis toxin inhibited the ability of this receptor to activate ERK. Finally, flow cytometry was utilized to assess the rate and extent of receptor internalization following agonist challenge. hPAR(2)C361A displayed faster internalization kinetics following trypsin activation compared with wt hPAR(2), whereas SLIGKV had a negligible effect on internalization for either receptor. In conclusion, palmitoylation plays an important role in the regulation of PAR(2) expression, agonist sensitivity, desensitization and internalization. PMID- 21627586 TI - Gem formation upon constitutive Gemin3 overexpression in Drosophila. AB - Gems or 'Gemini of Cajal bodies' are spherical nuclear aggregates of SMN (survival of motor neurons) complexes that frequently overlap Cajal bodies. Although described and characterized in mammalian tissues, gems have not been reported in invertebrates. Stimulation of gem formation in the fruitfly Drosophila melanogaster was investigated through the constitutive overexpression of a fluorescently tagged transgene of a DEAD-box SMN complex member, Gemin3, in wild-type tissues. Although expression was predominantly cytoplasmic in the larval brain cells, Gemin3 was found enriched in multiple discrete bright foci in the nuclei of several tissues including epidermis, muscle and gut. Similar to their mammalian counterparts, Drosophila gems contained endogenous SMN and at times overlapped with Cajal bodies. These findings support the hypothesis that gems are storage sites for excess nuclear SMN complexes and their frequent association with Cajal bodies might imply recruitment for nuclear ribonucleoprotein assembly reactions. PMID- 21627592 TI - [The use of 90% -3 polyunsaturated fatty acids for the treatment and prevention of cardiovascular diseases]. PMID- 21627593 TI - [Factors determining clinical effectiveness of clopidogrel and prognosis of patients with stable ischemic heart disease]. AB - Aim of the study was to elucidate genetic and drug factors affecting efficacy of clopidogrel in patients with ischemic heart disease - inhabitants of central region of Russian Federation. We included 399 patients with IHD (79% men, mean age 58.3+/-9 years) receiving long term therapy with clopidogrel 75 mg/day (during stable manifestations of the disease) or 75-150 mg/day in combination with aspirin (in relation with recent elective percutaneous interventions). We studied carriage of polymorphisms of genes controlling intestinal absorption of clopidogrel (ABCB1 C3435T), activation of clopidogrel in the liver (CYP2C19 *1 *2), and also registered concomitant administration of proton pump inhibitors (PPI). Then we determined relationship of these factors to development of vascular complications (vascular death/myocardial infarction/requirement in revascularization) during 18 months followup. Among studied genetic factors carriage of allele variants CYP2C19 *1/*2 and *2/* (found in 25.5 and 1.8% of patients, respectively), possessed prognostic significance. In the group of clopidogrel monotherapy carriage of at least one *2 allele was associated with increased rate of vascular complications (33.3% vs. 11.3%) including thrombotic complications (27.7% vs. 3.2%; =0.01). In patients receiving 75 mg/day of clopidogrel in combination with aspirin total rate of thrombotic complications as well as of all adverse unfavorable outcomes was higher in *2 carriers compared with wild type homozygotes (14.0% vs.8.7% and 21.0% vs. 15.8%, respectively). In patients receiving double dose clopidogrel in combination with aspirin we found no worsening of outcomes associated with CYP2C19*2 carriage. In the multifactorial risk model independent predictors of vascular complications turned out to be CYP2C19 *2/*2 homozygosity (RR 4.9; =0.02) and concomitant PPI administration ( 1.8; p=0.05). PMID- 21627594 TI - [Novel method of analysis of risk of development of ischemic heart disease with the use of genomic and computer technologies]. AB - AIM: to assess effectiveness of the use of modern methods of prognostication for assessment of risk of development of ischemic heart disease (IHD). MATERIAL AND METHODS: We examined 131 patients with diagnosis of IHD verified by coronary angiography and 159 subjects of control group. Initial information on each patient included the following parameters: traditional risk factors, laboratory parameters, results of instrumental examination, genetic markers. We studied 29 polymorphisms in 27 genes which according to international databases were associated with IHD. Genotype was assessed as 2 models: dominant and recessive. For each patient we calculated individual genetic index as sum of present polymorphic markers with addition of data of familial anamnesis. The data obtained were analyzed with the "RECOGNITION" system which used for solution of prognostication problems main approaches and algorithms of the theory of recognition by precedents. RESULTS: Accuracy of recognition varied from 70 to 75% with small number of traits and up to 90% on informative trait subsystems. The method "linear machine" showed the highest accuracy. The voting algorithm showed maximal accuracy of prognosis relative to some algorithms. In IHD prognostication most information systems comprised genetic markers, most significant of which was the genetic index representing sum of available polymorphic markers with addition of data of familial anamnesis. CONCLUSION: Analysis with the use of methods of recognition by precedents is a perspective technique for stratification of IHD risk and support of optimal decision making on prevention. The use of collectives of different methods of prognostication allows to increase accuracy of prognosis. PMID- 21627595 TI - [Prognostic value of levels of brain natriuretic peptide and genetic factors in patients after acute coronary syndrome]. AB - Prognostication of the course of disease in patients with high risk of unfavorable outcome of ischemic heart disease (IHD) is of great importance for creation of individualized strategy of treatment. We have investigated contribution of levels of brain natriuretic peptide (BNP) and genetic factors in the risk of development of complications of atherosclerosis in patients who have had acute coronary syndrome. We started to follow 324 patients on day 10 of stable state after acute coronary syndrome (55.1% with Q-wave myocardial infarction, 18.5% with non-Q myocardial infarction, 25.5% with unstable angina, men BNP level 624.5+/-32.13 mol/ml [70.3 - 4276.6]). Duration of followup was 2 years. Baseline BNP level in patients with unfavorable outcome during followup (fatal and nonfatal myocardial infarction and stroke) was 872.47+/-91.42 compared with 592.45+/-35.97 mol/ml in patients without unfavorable outcome (p=0,001). Multifactorial Cox analysis showed that carriage of T allele of polymorphic marker (--1654) of protein C gene, elevated BNP level, symptomatic atherosclerosis of peripheral arteries, history of MI, and use of thiazide diuretics were independently associated with unfavorable outcomes (p=0.026, <0.0001, <0.0001, =0.001, =0.024, respectively). Thus genetic factors and study of BNP allow to improve prediction of unfavorable outcome after exacerbation of IHD. PMID- 21627596 TI - [Echocardiographic assessment of myocardial fibrosis in young men with arterial hypertension and different types of left ventricular remodeling]. AB - In order to study structural functional characteristics of myocardium including parameters of myocardial fibrosis according echocardiography data in men with various levels of arterial pressure (AP) we examined 215 men aged 18-25 (mean 21.1+/-0,1) years with history of elevated AP at casual measurement. AP phenotype (normotension, stable arterial hypertension [AH], unstable AH) was determined on the basis of multiple measurements of clinical AP and 24 hour AP monitoring. At echocardiography we assessed presence of left ventricular (LV) hypertrophy (LVH), type of LV geometry, proportionality of LV myocardial mass (LVMM), diastolic function. Myocardial fibrosis was assessed by pixel density distribution range (PDDR) with the use of analysis of reflected signal. There were no manifestations of LV remodeling in subjects with normal AP. Concentric LV remodeling was found in 27.5 and 60.5% of patients with unstable and stable AH, respectively. Concentric LVH was found only in patients with stable AH (4.8%). Disproportionally high LVMM was found in 16.1% of subjects with stable AH. In a combined group with concentric LV remodeling and LVH rate of disproportionally high LVMM was 20.8%. We noted significant (p<0.001) increase of PDDR in stable AH (181.4+/-2.2) compared with PDDR in normal AP (164.6+/-4.6) and unstable AH (160.1+/-2.7). In stable I degree AH PDDR (177.3+/-2.2) was insignificantly lower than in II degree AH (185.7+/-3.9). PDDR in concentric LV remodeling was 180.5+/ 2.3, in concentric LVH- 166.8+/-13.2, in normal LV geometry - 168.4+/-2.5. PDDR in disproportionally high LVMM was higher than in proportional LVMM. Independent interrelationship was found between PDDR and body mass index (r=0.17; p=0.03), duration of AH (r=0.17; p=0.03), isovolumic relaxation time (r= 0.15; p=0.04). In young men LV remodeling can be detected at the stage of unstable AH. In stable AH degree of myocardial fibrosis was associated with higher AP level, concentric LV geometry, disproportionally high LVMM, lowering of diastolic function. PMID- 21627597 TI - [Clinico-statistical analysis of arterial hypertension complicated with hypertensive crisis in Moscow in 2005-2009]. AB - Clinicostatistical analysis of arterial hypertension complicated with hypertensive crisis using data of Moscow A.S.Puchkov Station of Urgent and Emergent Medical Aid revealed 14% rise in number of hypertensive crises during the period from 2005 to 2009. Number of hypertensive crises increased among persons of young age (18-35 years). Frequency of cerebrovascular complications of hypertensive crises was age dependent with maximal values among men aged 36-74 years and women older than 75 years. PMID- 21627598 TI - [Acute management at the rising of arterial pressure which is not menacing to a life]. AB - 491 hypertensive (non life threatening) patients were examined for hemodynamic changers and variance of the sinus rhythm after administration of clonidine, nifedipine, captopril, and mocsonidine. BP was done by the caff method, hemodynamic by the integral rheography, variance of the sinus rhythm - according to the recommendations of the International ECG Society. It was showed nifedipine and captopril were most effective among the others in case of moderate hypertension without sympathetic hyperfunction. In patients with severe and moderate hypertension follow up with the hyperfunction of the sympathetic nervous system clonidine and mocsonidine showed the major efficacy. In comparison to clonidine, mocsidimin is the medication which is not on the list, and beyond that it provided less adverse effects. PMID- 21627599 TI - [Clinical and angiographical factors influencing improvement of left ventricular function after implantation of dare-metal stents in patients with chronic total occlusions of coronary arteries]. AB - INTRODUCTION: Effect on left ventricular (LV) contractility of percutaneous coronary interventions (PCI) with implantation of bare-metal stents in patients with chronic total occlusions (CTO) of coronary arteries (CA) has not been completely studied. Aim of this study was to assess effect of PCI with implantation of bare-metal stents ("L+", Relisys, U) on LV ejection fraction (EF) and to investigate clinical and angiographical factors capable to affect restoration of LVEF. MATERIAL AND METHODS: We included in this study 154 patients after successful implantation of bare-metal stents in CTO of main epicardial CA. For assessment of LV function we performed echocardiographical examination before and in 6 months after PCI. RESULTS: Significant increase of LV EF (from 50.4 +/- 10.7 to 56.1+/-11.3%, p<0.0001), decreases of LV end diastolic (from 86.2+/-17.9 to 80,8+/-18,1 ml/m2, p<0.001) and end systolic (from 41.4+/-14.9 t 34.7+/-13.8 ml/m2 (p<0,001) volumes took place after implantation of stents. Multivariate analysis showed that initial LVEF <50%, duration of occlusion <2 months and absence of diabetes mellitus were independent predictors of improvement of LVEF. CONCLUSION: Implantation of bare-metal stents in patients with CTO CA affects positively LVEF during first 6 months after PCI especially in patients with lowered LVEF, in patients without diabetes mellitus, and duration of occlusion less or equal 2 months. PMID- 21627601 TI - [Adaptation reserve key concept for analysis of cardiological syndromes]. PMID- 21627600 TI - [Depressive symptoms worsen cardiovascular prognosis and shorten length of life in patients with arterial hypertension and ischemic heart disease]. AB - The KOORDINATA study (clinical epidemiological program of investigation of depression in cardiological practice in patients with arterial hypertension and ischemic heart disease) was a prospective 3 year long multicenter study of effects of depressive and anxiety states on the course and prognosis of ischemic heart disease (IHD) and arterial hypertension (AH). MATERIAL AND METHODS: Patients (n=5038, age more or equal 55 years) with verified IHD and/or AH from 37 cities were included into the study. Symptoms of anxiety and depression were assessed by the HADS scale (Hospital Anxiety And Depression Scale) validated in Russia. RESULTS: Clinically significant symptoms of anxiety ( more or equal 11 HADS) were present in 33 and 38%, of depression - in 30 and 38% of patients with IHD and AH, respectively. Presence of clinically manifested depression at initial examination increased 1.59 times combined risk of nonfatal and fatal cases (odds ratio [OR] 1.59, 95% confidence interval [CI] 1.30-1.96, p=0.0001), 1.64 times - risk of cardiovascular death (OR 1.64, 95% CI 1.20-2.24, p=0.0021), and 1.82 times - risk of death from all causes (OR 1.82, 95% CI 1.41-2.34, p=0.0001). It is known that for somatic patients crossover of anxiety and depression symptoms is typical (simultaneous presence of some symptoms of anxiety and depression). Total HADS score equal to 11 or more increased risk of all cause death (OR 1.41, 95% CI 1.08-1.84, p=0.0111), as well as fatal and nonfatal events (OR 1.33, 95% CI 1.08-1.65, p=0.0089). CONCLUSION: Taking into consideration negative effect of depressive symptoms on prognosis it is recommended to screen patients with IHD in the first place those who have had myocardial infarction and brain stroke with the aim of detection of depression and anxiety depressive symptoms and subsequent their adequate correction. PMID- 21627602 TI - [New platelet P2Y12 inhibitors versus clopidogrel in percutaneous coronary interventions. a meta-analysis]. PMID- 21627603 TI - [Anatomical, functional, and genetic peculiarities of myocardial hypertrophy in athletes]. AB - "Athletes Heart" (AH) is a complex of structural, electrophysiological, and functional changes - a consequence of physiological adaptation to regular physical exercise. AH is characterized by biventricular hypertrophy of ventricular myocardium at the background of unchanged parameters of systolic and diastolic function and depends on the type of exercise, body dimensions, sex, and genotype. Although degree of hypertrophy in AH in most cases does not exceed generally accepted limits problems of differential diagnostics of AH and other cardiovascular diseases manifesting by myocardial hypertrophy appear to be most actual. Contemporary possibilities of molecular genetic methods for detection of cause of myocardial hypertrophy are described. PMID- 21627604 TI - [Peculiarities of morphology and mechanisms of pathogenesis of hypertrophic cardiomyopathy]. AB - Understanding the molecular basis and cell mechanisms, clinical course, and treatment of hypertrophic cardiomyopathy (HCMP) has progressed substantially in the last decade. The majority of genetic mutations associated with HCMP occur in genes encoding sarcomeric proteins, which are expressed only in cardiomyocytes. The spectrum of morphological features of HCMP includes: hypertrophy of myocardium, myocardial disarray, interstitial fibrosis, mitral valve abnormalities, and microvascular remodeling, is indicative of the involvement of other cell lineages. The link between sarcomeric gene defects and these HCM phenotypes remains elusive. Based on novel insights provided by cardiac developmental biology we can create new effective methods of treatment of this complex disease. PMID- 21627605 TI - [Catheter ablation for atrial fibrillation: clinically established or still an experimental method]. AB - Interventional treatment for atrial fibrillation has been introduced as a therapeutic option since the pulmonary veins (PV) have been discovered as the dominant sources of paroxysmal atrial fibrillation (PAF). Elimination of PV conduction is the initial goal during catheter ablation in this setting. The success rate after the initial procedure varies between 60 and 85 %, with more than 80 % after subsequent interventions. Supported by the current guidelines, interventional treatment of AF is indicated in case of symptomatic arrhythmias refractory to antiarrhythmic treatment. The introduction of the combined, stepwise approach has been another important breakthrough with regard to the treatment of chronic persistent atrial fibrillation (CAF). This strategy includes the combination of all conventional ablation strategies (PV isolation, ablation of complex fractionated atrial electrograms, linear ablation) with the goal of AF termination. The first procedure for CAF treatment is quite frequently also only the first step towards stable sinus rhythm with a favourable outcome after AF termination (> 80 % sinus rhythm). In more than half of the patients predominantly other atrial arrhythmias than AF have to be targeted in a second procedure. This approach is currently under clinical investigation and so far not "clinically established" due to the fact that it is a quite time consuming and challenging procedure even in experienced centres. Future studies may help to identify predictors for procedure failure (e.g. LA size, AF duration, atrial cycle length, spectral analysis) in order to improve patient selection. Additionally, it has to be underscored, that in paroxysmal atrial fibrillation the relatively high recurrence rate after the first procedure still is the subject of further investigations. This aspect might be improved by the introduction of novel strategies (i.e. testing of concealed PV conduction after ablation with adenosine) or new technologies (i.e. robotic navigation) for PAF ablation. PMID- 21627606 TI - [Acute coronary syndrome: the system of organization of treatment]. PMID- 21627607 TI - [Myocardial infarction and stress at work place and in the family: 10-year risk of development in an open population of 2564 year old men (epidemiological study in a framework of the WHO program MONICA-PSYCHOSOCIAL)]. AB - With the aim of assessment of 10 year risk of myocardial infarction (MI) development in men in dependence on stress at work place and in the family in a framework of the WHO program "MONICA-PSYCHOSOCIAL" we examined random representative sample of men (n=657) aged 25-64 years inhabitants of one of districts in Novosibirsk. During 10 years (1994-2004) we registered all cases of MI. For assessment of relative risk of MI development we used COX proportional regression model. Among men with first MI 47.4% had high level of stress in the family and 57.9% were subjected to stress at work place. Five and 10 year risk of MI in men with high test levels of stress at home and work place was 2-5 times greater than in those without. Among men subjected to stress in the family MI risk was the highest at age 55-64 years, while permanent stressful situations at work place were more common in age group 45-54 years. Among men with high levels of stress in the family and at work place higher rate of MI development was observed in widowers, divorced men with incomplete high or elementary education, heavy or moderate manual labor workers, and pensioners. The results indicate that critical life events as well as chronic stressful influences increase risk of MI among men aged 25-64 years. The group of greatest risk - middle and old age persons who are less protected against social, political and economical disbalance in the society. PMID- 21627608 TI - [Prediction of left ventricular remodeling after acute myocardial infarction]. AB - Prediction of adverse left ventricular remodeling after acute myocardial infarction (AMI) may have important clinical implications. In this study, we evaluated the incidence of unfavorable changes of left ventricular volumes and ejection fraction after AMI. The ability of several cardiac markers, electrocardiographic and echocardiographic (including tissue Doppler) variables to predict adverse remodeling was demonstrated. The prognostic algorithm based on the investigated variables may help improve the treatment of patients after AMI. PMID- 21627609 TI - [Prognostic value of various markers of inflammation in ST-elevation myocardial infarction]. AB - We measured levels of several parameters of immune inflammation (interleukins [IL] 1, 6, 8, 10, 12, tumor necrosis factor [TNF], serum neopterin, C-reactive protein) on days 10-14 after onset of ST segment elevation myocardial infarction (MI) and related them to results of 12 months follow up. Elevated titers of IL-12 and neopterin were significantly associated with unfavorable events (repetitive MI, progression of angina, stroke, decompensation of chronic heart failure) (=0.001 and =0.003, respectively). After adjustment for initial severity of MI only IL-12 retained its prognostic value (=0.05). Regression analysis identified IL-12 level >108.7 g/ml and TIMI score > 3 as independent predictors of combined end point formation. Prognostic value of IL-12 level was significant both among patients with TIMI scores <3 and more or equal 4. PMID- 21627610 TI - [Lornoxicam for prevention of myocardial injury during acute ST-elevation myocardial infarction]. AB - 10% of patients with acute ST-elevation myocardial infarction (STEMI) treated with reperfusion therapy fail to develop an enzyme rise, but do exhibit transient ECG changes, which are consistent with an aborted myocardial infarction. Following reperfusion by primary PCI in STEMI, oxidative stress and an inflammatory response are induced immediately. Inflammation is a critical component of STEMI. Both COX isoforms are involved in reperfusion and ischemic myocardial injury. To evaluate the effectiveness of lornoxicam - nonselective NSAID, in decrease of myocardial injury during acute ST-elevation myocardial infarction. We analyzed 22 patients with STEMI, 14 of them received 16 mg and 8 mg lornoxicam after 20 min and 8 hours, respectively, after arrival to hospital. 12 f them received alteplase, 10 patients with cardiac pain up to 24 hours from the beginning, did not receive reperfusion therapy. All patients received anticoagulants, antiplatlet therapy, -blockers. The primary end point was all cause mortality by the day 30 and hospitalization due to congestive heart failure by the 1st year. There was no difference in mortality and heart failure by the 30 day and 1st year respectively, between the patients with STEMI treated with lornoxicam or placebo. Randomized controlled trials are needed to explore potential cardioprotective effects of lornoxicam in patients with acute STEMI. PMID- 21627611 TI - [The ORIGINAL program main results]. PMID- 21627612 TI - [Key laboratory diagnostic biomarkers of coronary atherosclerosis]. AB - Laboratory lipid and lipoprotein biomarkers (total cholesterol - CH, triglycerides - TG, low-density and high-density lipoprotein cholesterol- LDL-CH, HDL-CH, apolipoproteins B and A1 - apoB, apoA1), carbohydrate biomarkers (plasma glucose, basal insulin), high sensitive C-reactive protein (hsCRP) and oxidative biomarkers (basal level of lipid peroxidation [LPO] products in LDL, LDL resistance to oxidation in vitro, oxidative modification of apoLDL and level of LDL lipophilic antioxidants) were studied in 388 men aged 42-70 years: 96 citizens of Western Siberia with angiographically documented coronary atherosclerosis and coronary heart disease (CHD); 292 men of population sample of citizens of Novosibirsk, including 44 men with CHD confirmed by standardized criteria and methods. Significant associations were found of coronary atherosclerosis and CHD with laboratory diagnostic biomarkers like blood levels of HDL-CH, TG, apoB, apoA1, basal insulin, hsCRP and basal level of LPO products in LDL and LDL resistance to oxidation. PMID- 21627613 TI - [Changes of content of regulatory lymphocytes and concentration of soluble interleukine-2 receptor in blood of patients with ischemic heart disease after coronary artery angioplasty with implantation of stents with rapamycin covering]. AB - We studied dynamics of content of subpopulation of lymphocytes including regulatory and effector T-lymphocytes as well as concentration of soluble form of interleukine-2 receptor (sCD25) in peripheral blood of patients after coronary stenting (CS) with implantation of stents with rapamycin covering (SRC). We included into the study 62 patients with stable effort II-III functional class angina. Coronary angiography (CA) was carried out in all, CS with implantation of 1 - 2 SRC - in 42 patients. Blood samples were taken before CA/CS, in 24, 48 hours, 7 days, 1 and 3 months after intervention. Content of T-, helper and cytotoxic T-cells, -, NK-, NKT-cells, activated effector T-lymphocytes (CD4+CD251owCD127high) and regulatory T-lymphocytes (CD4+CD25highCD1271ow) were measured by direct immunofluorescence and flow cytometry. CD4+ lymphocytes were isolated from mononuclear cell fraction of donor blood by magnetic separation. Content of regulatory T-lymphocytes in culture were determined by expression of a specific marker FOXP3+. Concentration of sCD25 was measured by chemiluminescent method. It was shown that content of main subpopulations of lymphocytes in blood changed after CS or CF. Blood content of regulatory T-lymphocytes and sCD25 significantly increased after 7 days and 1 month after CS but not after CA. Plasma sCD25 concentration correlated with content of regulatory T-lymphocytes in 1 month after SRC implantation. During cultivation of CD4+ lymphocytes in the presence of rapamycin we noted antiproliferative effect relative to FOXP3-cells and accumulation of regulatory +-lymphocytes. Thus implantation of SRC in coronary arteries leads to increase of number of circulating regulatory T lymphocytes and blood concentration of sCD25. Changes of these parameters after CS can reflect peculiarities of local and systemic reaction arising in response to introduction of stent with drug covering and be significant for assessment of prognosis of the disease. PMID- 21627614 TI - [Adrenoreactivity in patients with arrhythmic syndrome associated with connective tissue dysplasia at the background of intake of the magnezium orotate]. AB - Elevated level of -adrenoreception of membranes is observed in patients with disturbances of heart rhythm at the background of connective tissue dysplasia. It is not related to characteristics of the arrhythmic syndrome. At the background of 4-months intake of preparation magnezium orotate significant decrease of degree of desensitization of of erythrocytes has been noted. PMID- 21627615 TI - [Association of C677T gene polymorphism of methylenetetrahydrofolate reductase with insulin resistance among Kirghizes]. AB - AIM: To study an association of C677T gene polymorphism of methylenetetrahydrofolate reductase (MTHFR) and insulin resistance (IR) among ethical Kirghizes. METHODS: 132 Kirghiz patients with IR according to HOMA index (n=132) and sex and age matched patients without IR, diabetes mellitus (DM) type 2 or metabolic syndrome (MS) (n=132) were included into this study. Measurements of blood pressure (BP), body mass index, waist and hip circumference, fasting blood sugar, insulin, lipid parameters and C677T gene polymorphism of MTHFR were performed in all patients. RESULTS: Frequency of CT and TT genotypes was significantly higher in patients with IR than in controls (2 - 7.22, p - 0,027, OR - 1.68, 95% confidence interval 1.13-2.5, p=0.027). T677 allele was also associated with obesity, hypertriglyceridemia and low level of high density lipoprotein cholesterol (HDL-C). CONCLUSION: In Kirghizes carriage of T677 allele of MTHFR gene was associated with IR, abdominal obesity, hypertriglyceridemia and low HDL-C level. PMID- 21627616 TI - [Diagnosis and classification of mitral valve prolapse in children and adolescents]. AB - We examined 1734 children and adolescents with admittance diagnosis mitral valve prolapse (MVP) and confirmed its presence using accepted Framingham criteria in 32.3% of cases. Reason for overdiagnosis in other cases was incorrect interpretation of auscultation or echocardiography data. Most often MVP was familial with recessive or dominant inheritance according to genealogical analysis. Contrary to MVP in adults children and adolescents commonly had no myxomatous transformation of valves. Taking into account constitutional and physiological peculiarities of the body in children and adolescents we distinguished two forms of MVP characteristic for this age: MVP as a variant of asthenic constitution and physiological MVP. Both forms are widely spread and represent normal states for juvenile population. PMID- 21627617 TI - [Hyperuricemia in chronic heart failure]. AB - Development of hyperuricemia is associated with excessive body mass, insulin resistance, metabolic syndrome, overuse of diuretics, elderly age, and abnormal renal function. Data are accumulated on existence of links between elevated uric acid level and arterial hypertension, diabetes mellitus, ischemic heart disease, and chronic heart failure (CHF). Hyperuricemia has been found in 60% of patients hospitalized because of decompensation of CHF. In CHF isolated hyperuricemia (irrespective of the state of renal function and administration of drugs) appears to be a marker of altered oxidative metabolism characterized by elevation of levels of free radicals which damage cardiomyocytes and vascular endothelium inducing disturbances of myocardial contractility and vasoconstriction. Hyperuricemia associated with insulin resistance, tissue hypoxia, elevated production of cytokines and free radicals can negatively affect cardiovascular system and worsen prognosis in patients with CHF. PMID- 21627618 TI - [Permanent cardiac pacing in vasovagal syncope and carotid sinus syndrome]. AB - Vasovagal syncope and carotid sinus syndrome are common conditions in young and elderly people, respectively, mostly with benign prognosis. Nevertheless, severe or "malignant" syncopal attacks in some patients may be associated with life threatening injury. Unfortunately, up to now almost all drug trials have failed to demonstrate any benefit in preventing syncope and interventional approach (pacemaker) may be appropriate. This article contains literature review and discussion of indications for pacing in vasovagal syncope and carotid sinus syndrome. PMID- 21627619 TI - [Utility of the use of computer monitoring pulse oximetry for screening of obstructive sleep apnea]. AB - This paper contains description of possibilities of the use of the computer monitoring pulse oximetry for (CMPO) diagnosis of obstructive sleep apnea syndrome at the level of practical health care and presentation of data on sensitivity and specificity of this method in various groups of patients. It is shown that diagnostic accuracy of the method substantially increases if patients are selected on the basis of clinical picture and presence of OSAS markers. A conclusion is made that CMPO can be successfully used for screening of respiration disturbances during sleep in patients with high probability of OSAS. A double stage scheme of patients screening is suggested. It assumes formation of a group with risk of development of OSAS basing on anamnesis, results of examination, and presence of concomitant conditions with subsequent CMPO. Criteria of assessment of results of CMPO according to value of desaturation index value are given. These criteria allow to more accurately predict presence or absence of OSAS. PMID- 21627620 TI - [Nitrous oxide and atherosclerosis. Pharmacotherapy of atherosclerosis and nitrous oxide]. PMID- 21627621 TI - The involvement of distinct neural systems in patients with obsessive-compulsive disorder with autogenous and reactive obsessions. AB - OBJECTIVE: To investigate the regional metabolite abnormalities and changes after treatment in patients with OCD with autogenous and reactive obsessions. METHOD: We assessed right anterior cingulate cortex (ACC) and amygdala-hippocampal region (Am + Hpp) N-acetyl-aspartate (NAA), choline (Cho) and creatine (Cr) concentrations and NAA/Cr and Cho/Cr ratios using single-voxel proton magnetic resonance spectroscopy in 15 patients with autogenous obsessions (OCD-A), 15 patients with reactive obsessions (OCD-R) and 15 healthy controls (HC). Measurements were repeated after 16 weeks of fluoxetine treatment. RESULTS: Baseline ACC NAA/Cr ratios of both OCD groups were significantly lower than HC. OCD-A group had significantly lower baseline NAA/Cr ratios in the Am + Hpp than other groups. These differences were more likely to be explained by higher Cr levels in ACC. We found no significant differences and changes for Cho levels and Cho/Cr ratios between groups and within groups. Significant increase in NAA/Cr ratios of OCD-A group found in the Am + Hpp was more likely to be explained by increased NAA levels. No significant changes were found in ACC NAA/Cr ratios. CONCLUSION: While disturbed energy metabolism in ACC might reflect a common pathology in patients with OCD regardless of symptom dimension, alterations in mesiotemporal lobe are more likely for autogenous obsessions. PMID- 21627622 TI - Stellate ganglion blockade for analgesia following upper limb surgery. AB - We report the successful use of a stellate ganglion block as part of a multi modal postoperative analgesic regimen. Four patients scheduled for orthopaedic surgery following upper limb trauma underwent blockade of the stellate ganglion pre-operatively under ultrasound guidance. Patients reported excellent postoperative analgesia, with postoperative VAS pain scores between 0 and 2, and consumption of morphine in the first 24 h ranging from 0 to 14 mg. While these are preliminary findings, and must be confirmed in a clinical trial, they highlight the potential for stellate ganglion blockade to provide analgesia following major upper limb surgery. PMID- 21627623 TI - The role of the autonomic nervous system in acute surgical pain processing - what do we know? PMID- 21627624 TI - Case reports of novel treatments - proper evaluation before clinical use. PMID- 21627625 TI - Time for capnography - everywhere. PMID- 21627626 TI - Bayesian modeling for genetic anticipation in presence of mutational heterogeneity: a case study in Lynch syndrome. AB - Genetic anticipation, described by earlier age of onset (AOO) and more aggressive symptoms in successive generations, is a phenomenon noted in certain hereditary diseases. Its extent may vary between families and/or between mutation subtypes known to be associated with the disease phenotype. In this article, we posit a Bayesian approach to infer genetic anticipation under flexible random effects models for censored data that capture the effect of successive generations on AOO. Primary interest lies in the random effects. Misspecifying the distribution of random effects may result in incorrect inferential conclusions. We compare the fit of four-candidate random effects distributions via Bayesian model fit diagnostics. A related statistical issue here is isolating the confounding effect of changes in secular trends, screening, and medical practices that may affect time to disease detection across birth cohorts. Using historic cancer registry data, we borrow from relative survival analysis methods to adjust for changes in age-specific incidence across birth cohorts. Our motivating case study comes from a Danish cancer register of 124 families with mutations in mismatch repair (MMR) genes known to cause hereditary nonpolyposis colorectal cancer, also called Lynch syndrome (LS). We find evidence for a decrease in AOO between generations in this article. Our model predicts family-level anticipation effects that are potentially useful in genetic counseling clinics for high-risk families. PMID- 21627627 TI - A shrinkage approach for estimating a treatment effect using intermediate biomarker data in clinical trials. AB - In clinical trials, a biomarker (S ) that is measured after randomization and is strongly associated with the true endpoint (T) can often provide information about T and hence the effect of a treatment (Z ) on T. A useful biomarker can be measured earlier than T and cost less than T. In this article, we consider the use of S as an auxiliary variable and examine the information recovery from using S for estimating the treatment effect on T, when S is completely observed and T is partially observed. In an ideal but often unrealistic setting, when S satisfies Prentice's definition for perfect surrogacy, there is the potential for substantial gain in precision by using data from S to estimate the treatment effect on T. When S is not close to a perfect surrogate, it can provide substantial information only under particular circumstances. We propose to use a targeted shrinkage regression approach that data-adaptively takes advantage of the potential efficiency gain yet avoids the need to make a strong surrogacy assumption. Simulations show that this approach strikes a balance between bias and efficiency gain. Compared with competing methods, it has better mean squared error properties and can achieve substantial efficiency gain, particularly in a common practical setting when S captures much but not all of the treatment effect and the sample size is relatively small. We apply the proposed method to a glaucoma data example. PMID- 21627628 TI - Split-plot designs for robotic serial dilution assays. AB - This article explores effective implementation of split-plot designs in serial dilution bioassay using robots. We show that the shortest path for a robot to fill plate wells for a split-plot design is equivalent to the shortest common supersequence problem in combinatorics. We develop an algorithm for finding the shortest common supersequence, provide an R implementation, and explore the distribution of the number of steps required to implement split-plot designs for bioassay through simulation. We also show how to construct collections of split plots that can be filled in a minimal number of steps, thereby demonstrating that split-plot designs can be implemented with nearly the same effort as strip-plot designs. Finally, we provide guidelines for modeling data that result from these designs. PMID- 21627629 TI - Estimating effect sizes of differentially expressed genes for power and sample size assessments in microarray experiments. AB - In microarray screening for differentially expressed genes using multiple testing, assessment of power or sample size is of particular importance to ensure that few relevant genes are removed from further consideration prematurely. In this assessment, adequate estimation of the effect sizes of differentially expressed genes is crucial because of its substantial impact on power and sample size estimates. However, conventional methods using top genes with largest observed effect sizes would be subject to overestimation due to random variation. In this article, we propose a simple estimation method based on hierarchical mixture models with a nonparametric prior distribution to accommodate random variation and possible large diversity of effect sizes across differential genes, separated from nuisance, nondifferential genes. Based on empirical Bayes estimates of effect sizes, the power and false discovery rate (FDR) can be estimated to monitor them simultaneously in gene screening. We also propose a power index that concerns selection of top genes with largest effect sizes, called partial power. This new power index could provide a practical compromise for the difficulty in achieving high levels of usual overall power as confronted in many microarray experiments. Applications to two real datasets from cancer clinical studies are provided. PMID- 21627630 TI - A new criterion for confounder selection. AB - We propose a new criterion for confounder selection when the underlying causal structure is unknown and only limited knowledge is available. We assume all covariates being considered are pretreatment variables and that for each covariate it is known (i) whether the covariate is a cause of treatment, and (ii) whether the covariate is a cause of the outcome. The causal relationships the covariates have with one another is assumed unknown. We propose that control be made for any covariate that is either a cause of treatment or of the outcome or both. We show that irrespective of the actual underlying causal structure, if any subset of the observed covariates suffices to control for confounding then the set of covariates chosen by our criterion will also suffice. We show that other, commonly used, criteria for confounding control do not have this property. We use formal theory concerning causal diagrams to prove our result but the application of the result does not rely on familiarity with causal diagrams. An investigator simply need ask, "Is the covariate a cause of the treatment?" and "Is the covariate a cause of the outcome?" If the answer to either question is "yes" then the covariate is included for confounder control. We discuss some additional covariate selection results that preserve unconfoundedness and that may be of interest when used with our criterion. PMID- 21627631 TI - Dose-response curve estimation: a semiparametric mixture approach. AB - In the estimation of a dose-response curve, parametric models are straightforward and efficient but subject to model misspecifications; nonparametric methods are robust but less efficient. As a compromise, we propose a semiparametric approach that combines the advantages of parametric and nonparametric curve estimates. In a mixture form, our estimator takes a weighted average of the parametric and nonparametric curve estimates, in which a higher weight is assigned to the estimate with a better model fit. When the parametric model assumption holds, the semiparametric curve estimate converges to the parametric estimate and thus achieves high efficiency; when the parametric model is misspecified, the semiparametric estimate converges to the nonparametric estimate and remains consistent. We also consider an adaptive weighting scheme to allow the weight to vary according to the local fit of the models. We conduct extensive simulation studies to investigate the performance of the proposed methods and illustrate them with two real examples. PMID- 21627632 TI - NSAID gastropathy and enteropathy: distinct pathogenesis likely necessitates distinct prevention strategies. AB - The mechanisms underlying the ability of nonsteroidal anti-inflammatory drugs (NSAIDs) to cause ulceration in the stomach and proximal duodenum are well understood, and this injury can largely be prevented through suppression of gastric acid secretion (mainly with proton pump inhibitors). In contrast, the pathogenesis of small intestinal injury induced by NSAIDs is less well understood, involving more complex mechanisms than those in the stomach and proximal duodenum. There is clear evidence for important contributions to NSAID enteropathy of enteric bacteria, bile and enterohepatic recirculation of the NSAID. There is no evidence that suppression of gastric acid secretion will reduce the incidence or severity of NSAID enteropathy. Indeed, clinical data suggest little, if any, benefit. Animal studies suggest a significant exacerbation of NSAID enteropathy when proton pump inhibitors are co-administered with the NSAID. This worsening of damage appears to be linked to changes in the number and types of bacteria in the small intestine during proton pump inhibitor therapy. The distinct mechanisms of NSAID-induced injury in the stomach/proximal duodenum versus the more distal small intestine likely dictate distinct strategies for prevention. PMID- 21627633 TI - The orexin receptor OX(1)R in colon cancer: a promising therapeutic target and a new paradigm in G protein-coupled receptor signalling through ITIMs. AB - An exciting aspect of the heptahelical orexin receptor 1 (OX(1)R) has emerged recently, when it was shown that it drives apoptosis in human colon cancer cell lines. Here we review recent findings related to the role of OX(1)R in colorectal cancers and the unexpected mechanism whereby this G protein-coupled receptor works. The OX(1)R is aberrantly expressed at all steps of primary colorectal tumour progression and after local (lymph node) or distant (liver, lung) metastasis. No OX(1)R is detected in normal colonic epithelial cells. Treatment of human colon cancer cells in culture with orexins promotes robust apoptosis and subsequent reduction of growth including in cells that are resistant to 5 fluorouracil, the most commonly used drug in chemotherapy. When human colon cancer cells are xenografted in nude mice, treatment with orexins dramatically slows tumour growth and even reverses the development of established tumours. Thus, OX(1)R agonists might be novel candidates for colon cancer therapy. Activation of OX(1)R drives apoptosis through G(q) protein but independently of classical Galpha(q) activation of phospholipase C. In fact, it is the freed betagamma dimer of G(q) that plays a pivotal role by stimulating Src-tyrosine kinase. This results in phosphorylation of two immunoreceptor tyrosine-based inhibitory motifs (ITIM) in OX(1)R and subsequent recruitment by OX(1)R of the phosphotyrosine phosphatase SHP-2, which is activated thereby. Downstream events include release of cytochrome c from mitochondria and activation of caspase-3 and caspase-7. The role of ITIMs in OX(1)R-driven apoptosis represents a new paradigm of G protein-coupled receptor signalling. PMID- 21627634 TI - New concepts in calcium-sensing receptor pharmacology and signalling. AB - The calcium-sensing receptor (CaR) is the key controller of extracellular calcium (Ca(2+)(o)) homeostasis via its regulation of parathyroid hormone (PTH) secretion and renal Ca(2+) reabsorption. The CaR-selective calcimimetic drug Cinacalcet stimulates the CaR to suppress PTH secretion in chronic kidney disease and represents the world's first clinically available receptor positive allosteric modulator (PAM). Negative CaR allosteric modulators (NAMs), known as calcilytics, can increase PTH secretion and are being investigated as possible bone anabolic treatments against age-related osteoporosis. Here we address the current state of development and clinical use of a series of positive and negative CaR modulators. In addition, clinical CaR mutations and transgenic mice carrying tissue-specific CaR deletions have provided a novel understanding of the relative functional importance of CaR in both calciotropic tissues and those elsewhere in the body. The development of CaR-selective modulators and signalling reagents have provided us with a more detailed appreciation of how the CaR signals in vivo. Thus, both of these areas of CaR research will be reviewed. PMID- 21627635 TI - Systemic urocortin 2, but not urocortin 1 or stressin 1-A, suppresses feeding via CRF2 receptors without malaise and stress. AB - BACKGROUND AND PURPOSE: Infusion of corticotropin-releasing factor (CRF)/urocortin (Ucn) family peptides suppresses feeding in mice. We examined whether rats show peripheral CRF/Ucn-induced anorexia and determined its behavioural and pharmacological bases. EXPERIMENTAL APPROACH: Male Wistar rats (n= 5-12 per group) were administered (i.p.) CRF receptor agonists with different subtype affinities. Food intake, formation of conditioned taste aversion and corticosterone levels were assessed. In addition, Ucn 1- and Ucn 2-induced anorexia was studied in fasted CRF(2) knockout (n= 11) and wild-type (n= 13) mice. KEY RESULTS: Ucn 1, non-selective CRF receptor agonist, reduced food intake most potently (~0.32 nmol.kg(-1) ) and efficaciously (up to 70% reduction) in fasted and fed rats. The peptides' rank-order of anorexic potency was Ucn 1 >= Ucn 2 > >stressin(1) -A > Ucn 3, and efficacy, Ucn 1 > stressin(1) -A > Ucn 2 = Ucn 3. Ucn 1 reduced meal frequency and size, facilitated feeding bout termination and slowed eating rate. Stressin(1) -A (CRF(1) agonist) reduced meal size; Ucn 2 (CRF(2) agonist) reduced meal frequency. Stressin(1) -A and Ucn 1, but not Ucn 2, produced a conditioned taste aversion, reduced feeding efficiency and weight regain and elicited diarrhoea. Ucn 1, but not Ucn 2, also increased corticosterone levels. Ucn 1 and Ucn 2 reduced feeding in wild-type, but not CRF(2) knockout, mice. CONCLUSIONS AND IMPLICATIONS: CRF(1) agonists, Ucn 1 and stressin(1) -A, reduced feeding and induced interoceptive stress, whereas Ucn 2 potently suppressed feeding via a CRF(2) -dependent mechanism without eliciting malaise. Consistent with their pharmacological differences, peripheral urocortins have diverse effects on appetite. PMID- 21627636 TI - Opioids and the immune system: what is their mechanism of action? AB - There is a significant amount of literature showing that morphine and other opioids modulate immune responses. The findings support many mechanisms by which this may occur. In vitro experiments provide evidence for direct actions of opioids on immune cells using a variety of functional end points. When these drugs are given in vivo, a plethora of immune parameters are also altered. The paper in this issue of the journal by Zhang et al. provides new information on morphine alteration of immune cell subsets in the spleen and thymus of mice and the potential role of glucocorticoids in these observed phenomena. This Commentary reviews the in vitro activities of morphine on leucocytes, as well as other documented mechanisms by which morphine can alter immune function in vivo. PMID- 21627637 TI - Desmoteplase: discovery, insights and opportunities for ischaemic stroke. AB - Nature has provided a vast array of bioactive compounds that have been exploited for either diagnostic or therapeutic use. The field of thrombosis and haemostasis in particular has enjoyed much benefit from compounds derived from nature, notably from snakes and blood-feeding animals. Indeed, the likelihood that blood feeding animals would harbour reagents with relevant pharmacology and with potential pharmaceutical benefit in haemostasis was not too far-fetched. Blood feeding animals including leeches and ticks have evolved a means to keep blood from clotting or to at least maintain the liquid state, and some of these have been the subject of clinical development. A more recent example of this has been the saliva of the common vampire bat Desmodus rotundus, which has proven to harbour a veritable treasure trove of novel regulatory molecules. Among the bioactive compounds present is a fibrinolytic compound that was shown over 40 years ago to be a potent plasminogen activator. Studies of this vampire bat derived plasminogen activator, more recently referred to as desmoteplase, revealed that this protease shared a number of structural and functional similarities to the human fibrinolytic protease, tissue-type plasminogen activator (t-PA) yet harboured critically important differences that have rendered this molecule attractive for clinical development for patients with ischaemic stroke. PMID- 21627638 TI - Antiparkinsonian potential of targeting group III metabotropic glutamate receptor subtypes in the rodent substantia nigra pars reticulata. AB - BACKGROUND AND PURPOSE: Increased firing of the glutamatergic pathway between the subthalamic nucleus and substantia nigra pars reticulata (SNpr) contributes to the abnormal firing of motor circuits and subsequent motor deficits seen in Parkinson's disease. Broad spectrum agonist-induced activation of presynaptic group III metabotropic glutamate (mGlu) receptors within the SNpr reduced glutamate release and reversed akinesia in the reserpine-treated rat model of Parkinson's disease. Here, we have sought to identify which subtypes of group III mGlu receptor in the SNpr were responsible for these beneficial effects. EXPERIMENTAL APPROACH: The ability of the mGlu(4) positive allosteric modulator, N-phenyl-7-(hydroxyminocyclopropa[b]chromen-1a-carboxamide) (PHCCC), the mGlu(7) allosteric agonist, N,N'-dibenzhydrylethane-1,2-diamine dihydrochloride (AMN082) and the mGlu(8) -selective agonist (S)-3,4-dicarboxyphenylglycine [(S)-3,4-DCPG] to inhibit KCl-evoked [(3) H]-D-aspartate release was examined in vitro in rat nigral prisms. Reversal of akinesia in reserpine-treated rats was also assessed following intranigral injection of these agents. KEY RESULTS: PHCCC and AMN082 inhibited [(3) H]-D-aspartate release by 42% and 53%, respectively when given alongside a sub-threshold concentration of the broad spectrum group III agonist, L-2-amino-4-phosphonobutyrate (L-AP4; 1 uM). In contrast (S)-3,4-DCPG failed to inhibit [(3) H]-D-aspartate release. All three agents also reversed reserpine induced akinesia although only the effects of PHCCC and AMN082 were inhibited by pre-treatment with the group III antagonist (RS)-alpha-cyclopropyl-4 phosphonophenylglycine (CPPG). CONCLUSIONS AND IMPLICATIONS: These findings reveal that targeting SNpr mGlu(4) or mGlu(7) receptors, but not mGlu(8) receptors, provided relief from akinesia in the reserpine-treated rat model of Parkinson's disease, most likely reflecting inhibition of excess glutamate release in this region. PMID- 21627639 TI - Subchronic treatment with fluoxetine and ketanserin increases hippocampal brain derived neurotrophic factor, beta-catenin and antidepressant-like effects. AB - BACKGROUND AND PURPOSE: 5-HT(2A) receptor antagonists improve antidepressant responses when added to 5-HT-selective reuptake inhibitors (SSRIs) or tricyclic antidepressants. Here, we have studied the involvement of neuroplasticity pathways and/or the 5-hydroxytryptaminergic system in the antidepressant-like effect of this combined treatment, given subchronically. EXPERIMENTAL APPROACH: Expression of brain-derived neurotrophic factor (BDNF) and its receptor (TrkB), 5 bromo-2'-deoxyuridine (BrdU) incorporation, and beta-catenin protein expression in different cellular fractions, as well as 5-HT(1A) receptor function were measured in the hippocampus of rats treated with fluoxetine, ketanserin and fluoxetine + ketanserin for 7 days, followed by a forced swimming test (FST) to analyse antidepressant efficacy. KEY RESULTS: mRNA for BDNF was increased in the CA3 field and dentate gyrus of the hippocampus by combined treatment with fluoxetine + ketanserin. Expression of beta-catenin was increased in total hippocampal homogenate and in the membrane fraction, but unchanged in the nuclear fraction after combined treatment with fluoxetine + ketanserin. These effects were paralleled by a decreased immobility time in the FST. There were no changes in BrdU incorporation, TrkB expression and 5-HT(1A) receptor function in any of the groups studied. CONCLUSIONS AND IMPLICATIONS: The antidepressant-like effect induced by subchronic co-treatment with a SSRI and a 5-HT(2A) receptor antagonist may mainly be because of modifications in hippocampal neuroplasticity (BDNF and membrane-associated beta-catenin), without a significant role for other mechanisms involved in chronic antidepressant response, such as hippocampal neuroproliferation or 5-HT(1A) receptor desensitization in the dorsal raphe nucleus. PMID- 21627640 TI - Sildenafil restores cognitive function without affecting beta-amyloid burden in a mouse model of Alzheimer's disease. AB - BACKGROUND AND PURPOSE: Inhibitors of phosphodiesterase 5 (PDE5) affect signalling pathways by elevating cGMP, which is a second messenger involved in processes of neuroplasticity. In the present study, the effects of the PDE5 inhibitor, sildenafil, on the pathological features of Alzheimer's disease and on memory-related behaviour were investigated. EXPERIMENTAL APPROACH: Sildenafil was administered to the Tg2576 transgenic mouse model of Alzheimer's disease and to age-matched negative littermates (controls). Memory function was analysed using the Morris water maze test and fear conditioning tasks. Biochemical analyses were performed in brain lysates from animals treated with saline or with sildenafil. KEY RESULTS: Treatment of aged Tg2576 animals with sildenafil completely reversed their cognitive impairment. Such changes were accompanied in the hippocampus by a reduction of tau hyperphosphorylation and a decrease in the activity of glycogen synthase kinase 3beta (GSK3beta) and of cyclin-dependent kinase 5 (CDK5) (p25/p35 ratio). Moreover, sildenafil also increased levels of brain-derived neurotrophic factor (BDNF) and the activity-regulated cytoskeletal-associated protein (Arc) in the hippocampus without any detectable modification of brain amyloid burden. CONCLUSIONS AND IMPLICATIONS: Sildenafil improved cognitive functions in Tg2576 mice and the effect was not related to changes in the amyloid burden. These data further strengthen the potential of sildenafil as a therapeutic agent for Alzheimer's disease. PMID- 21627641 TI - Exome sequencing in a family segregating for celiac disease. AB - Celiac disease is a multifactorial disorder caused by an unknown number of genetic factors interacting with an environmental factor. Hence, most patients are singletons and large families segregating with celiac disease are rare. We report on a three-generation family with six patients in which the inheritance pattern is consistent with an autosomal dominant model. To date, 27 loci explain up to 40% of the heritable disease risk. We hypothesized that part of the missing heritability is because of low frequency or rare variants. Such causal variants could be more prominent in multigeneration families where private mutations might co-segregate with the disease. They can be identified by linkage analysis combined with whole exome sequencing. We found three linkage regions on 4q32.3 4q33, 8q24.13-8q24.21 and 10q23.1-10q23.32 that segregate with celiac disease in this family. We performed exome sequencing on two affected individuals to investigate the positional candidate regions and the remaining exome for causal nonsense variants. We identified 12 nonsense mutations with a low frequency (minor allele frequency <10%) present in both individuals, but none mapped to the linkage regions. Two variants in the CSAG1 and KRT37 genes were present in all six affected individuals. Two nonsense variants in the MADD and GBGT1 genes were also present in 5 of 6 and 4 of 6 individuals, respectively; future studies should determine if any of these nonsense variants is causally related to celiac disease. PMID- 21627642 TI - Exome sequencing and the genetics of intellectual disability. AB - Exome sequencing has greatly impacted the speed at which new disease genes are identified. In the last year alone, six studies have used exome sequencing to identify new genes involved in intellectual disability, a genetically heterogeneous condition affecting 1-3% of the population. These studies encompass the full gamut of modes of inheritance and phenotypic presentation, including syndromic and non-syndromic conditions, sporadic and familial cases, and dominant and recessive inheritance patterns. Because different disease presentations require different approaches to gene discovery, studies of intellectual disability provide a nearly comprehensive showcase of strategies for exome-driven gene discovery. Despite these successes, the etiology of ~60% of cases of intellectual disability remains unknown. The application of exome sequencing to the clinical diagnosis of intellectual disability in the near future will ultimately reduce the number of idiopathic cases and provide a rich source of sequence variation for the identification of new intellectual disability genes. PMID- 21627643 TI - Ictal head turning in frontal and temporal lobe epilepsy. AB - PURPOSE: To quantitatively evaluate the difference of ictal head turning movements between patients with temporal lobe epilepsy (TLE) and frontal lobe epilepsy (FLE). METHODS: We investigated 38 seizures of 31 patients with unilateral TLE and 22 seizures of 14 patients with unilateral FLE where head turning occurred in the seizure evolution. The head movements were defined as ipsilateral or contralateral in reference to the lateralization of the patient's focal epilepsy syndrome. Head movements were quantified by either referencing the head position with manually placed markers or by automatic detection of infrared marked reference points. The time of onset, duration, and angular speed of the head movements were computed, and interindividual and intraindividual analyses were performed. KEY FINDINGS: All of the TLE seizures had both contralateral and ipsilateral head turning, whereas all FLE had contralateral head turning; only 6 of 22 seizures were associated with ipsilateral head turning. Ipsilateral head turning always preceded contralateral head turning in both TLE and FLE. The head turning occurred significantly sooner after clinical seizure onset in FLE than in TLE patients (ipsilateral 0.5 vs. 16.0 s, contralateral: 4.5 vs. 21.3 s; p < 0.001). Furthermore, the duration of head turning was shorter in FLE for contralateral head turning (4.1 s) than in TLE (contralateral 6.0 s, p < 0.01); the ipsilateral head turning in the two groups did not differ (3.0 vs. 2.9 s) in duration. The angular speed of head turning did not differ for ipsilateral and for contralateral head turning in FLE and TLE. SIGNIFICANCE: Quantitative analysis of head turning demonstrates significant differences between patients with FLE and TLE. These differences likely represent differences in spread of epileptic activity. This information may be useful in the seizure evaluation of patients considered for resective epilepsy surgery. PMID- 21627644 TI - Targeting the endocannabinoid system in the amygdala kindling model of temporal lobe epilepsy in mice. AB - The endocannabinoid system can be considered as a putative target to affect ictogenesis as well as the generation of a hyperexcitable epileptic network. Therefore, we evaluated the effect of a CB1 receptor agonist (WIN55.212-2) and of an inhibitor of the enzymatic degradation of the endocannabinoid anandamide (fatty acid hydrolase inhibitor URB597) in the amygdala kindling model of temporal lobe epilepsy. Only minor effects on seizure thresholds and seizure parameters without a clear dose-dependency were observed in fully kindled mice. When evaluating the impact on kindling acquisition, WIN55.212-2 significantly delayed the progression of seizure severity. In contrast, URB597 did not affect the development of seizures in the kindling paradigm. Analysis of cell proliferation and neurogenesis during the kindling process revealed that URB597 significantly reduced the number of newborn neurons. These data give first evidence that CB1-receptor activation might render a disease-modifying approach. Future studies are necessary that further analyze the role of CB1 receptors and to confirm the efficacy of CB1-receptor agonists in other models of chronic epilepsy. PMID- 21627645 TI - Modulation of peripheral cytotoxic cells and ictogenesis in a model of seizures. AB - PURPOSE: A link between seizure susceptibility, blood-brain barrier (BBB) failure, and the activation of peripheral white blood cells has been recently proposed. However, the molecular players involved in this cascade of events are unknown. We tested the hypothesis that immunosupression by splenectomy or lack of perforin, a downstream factor of natural killer (NK) and cytotoxic T cells, could reduce seizure onset. METHODS: Pilocarpine was used to induce seizures in adult rats wild-type and perforin-deficient mice. Splenectomy was performed prior to pilocarpine injection. Seizure onset was evaluated by electroencephalography (EEG) and joint time-frequency analysis. Spleens from control and pilocarpine treated groups were analyzed for anatomical changes and CD3+ cell content. BBB damage was assessed by measuring albumin parenchymal extravasation. Fluorescence activated cell sorting (FACS) analysis was performed on spleen and brain tissue of wild-type and perforin-deficient mice treated, or not, with pilocarpine. KEY FINDINGS: Splenectomy significantly reduced seizure-associated mortality. Histologic analysis of the spleens exposed to pilocarpine revealed altered white and red pulp anatomy and an increase in CD3+ T cells. Onset of status epilepticus (SE) and mortality were significantly decreased in perforin-deficient mice. Pilocarpine significantly increased spleen NK 1.1 and CD8+ cell percentage; in contrast, the brain inflammatory cell profile remained unchanged at the time of pilocarpine SE. BBB damage was reduced in the perforin-deficient pilocarpine treated mice. SIGNIFICANCE: Immunosuppressant maneuvers such as splenectomy or lack of perforin decrease the onset or the severity of pilocarpine SE. Our results suggest that cytotoxic lymphocytes, and specifically the cytolytic factor perforin, may be key molecular players involved in the axis between peripheral intravascular inflammation and seizures. PMID- 21627646 TI - Aphasic status epilepticus: electroclinical correlation. AB - PURPOSE: Aphasic status epilepticus (ASE) in otherwise awake patients is a rare phenomenon. We present a series of nine consecutive patients with ASE to characterize clinical, electrophysiologic, and imaging findings. METHODS: Nine patients in ASE were identified between July 2006 and December 2009 at our institution. Each was evaluated by the neurology service and monitored with video electroencephalography (EEG) for at least 24 h. Thorough, repeated language testing was correlated with EEG findings. KEY FINDINGS: All nine patients were right-handed with subacute or chronic left hemispheric lesions on magnetic resonance imaging (MRI). All patients had mixed aphasia, three presenting with persistent aphasia from onset and six with episodic speech impairment, which became persistent in five of the six. The initial 30-min EEG demonstrated electrographic seizure in only five patients (56%), despite the presence of aphasia during the recording. Left hemispheric periodic lateralized epileptiform discharges (PLEDS) were seen in two patients, and left hemispheric slowing in two patients. Continuous video-EEG monitoring confirmed electrographic seizure activity in all nine patients. Peak electrographic seizure frequency varied from continuous to once every 2 h and was not associated with fluctuations in the speech deficit. EEG seizures resolved abruptly in three patients and gradually over up to 4 days in six patients. Clinical improvement was delayed in eight of the nine patients, and four patients retained some aphasia at discharge, 2-4 days after EEG seizure resolution. SIGNIFICANCE: Standard EEG is sensitive for detection of abnormalities in the dominant hemisphere in patients with ASE. However, continuous EEG is necessary to confirm the diagnosis and monitor treatment, since clinical symptoms do not correlate with electrographic seizure activity and do not provide sufficient information to guide treatment decisions. PMID- 21627647 TI - Widespread cortical thinning in children with frontal lobe epilepsy. AB - PURPOSE: Spread of seizure activity outside the frontal lobe due to cortico cortical connections can result in alteration in the cortex beyond the frontal lobe in children with intractable frontal lobe epilepsy (FLE). The aim of this study was to identify regions of reduced cortical thickness in children with intractable FLE. METHODS: High-resolution volumetric T(1)-weighted imaging was performed on 17 children with FLE, who were being evaluated for epilepsy surgery, and 26 age-matched healthy controls. The cortical thickness of 12 patients with left FLE and 5 patients with right FLE was compared to controls. The clusters of cortical thinning were regressed against age of seizure onset, duration of epilepsy, seizure frequency, and number of medications. KEY FINDINGS: In children with left FLE, cortical thinning was present in the left superior frontal, paracentral, precuneus, cingulate, inferior parietal, supramarginal, postcentral, and superior temporal gyri, as well as in the right superior and middle frontal, medial orbitofrontal, supramarginal, postcentral, banks of superior temporal sulcus, and parahippocampal gyri. In children with right FLE, cortical thinning was present in the right precentral, postcentral, transverse temporal, parahippocampal, lingual, and lateral occipital gyri, as well as in the left superior frontal, inferior parietal, postcentral, superior temporal, posterior cingulate, and lingual gyri. In children with left FLE, following exclusion of one outlier, there was no significant association between age at seizure onset, duration of epilepsy, seizure frequency and number of medications with clusters of cortical thinning. In children with right FLE, age at seizure onset, duration of epilepsy, frequency of seizures, and number of medications were not associated with clusters of cortical thinning within the right and left hemispheres. SIGNIFICANCE: Cortical changes were present in the frontal and extrafrontal cortex in children with intractable FLE. These changes may be related to spread of seizure activity, large epileptogenic zones involving both frontal and extrafrontal lobes, and development of secondary epileptogenic zones that over time lead to cortical abnormality. Further studies correlating cortical changes with neurocognitive measures are needed to determine if the cortical changes relate to cognitive function. PMID- 21627648 TI - Swine model for translational research of invasive intracranial monitoring. AB - Focal cortical epilepsy is currently studied most effectively in humans. However, improvement in cortical monitoring and investigational device development is limited by lack of an animal model that mimics human acute focal cortical epileptiform activity under epilepsy surgery conditions. Therefore, we assessed the swine model for translational epilepsy research. Swine were used due to their cost-effectiveness, convoluted cortex, and comparative anatomy. The anatomy has all the same brain structures as the human, and in similar locations. Focal subcortical injection of benzyl-penicillin produced clinical seizures correlating with epileptiform activity demonstrating temporal and spatial progression. Swine were evaluated under five different anesthesia regimens. Of the five regimens, conditions similar to human intraoperative anesthesia, including continuous fentanyl with low dose isoflorane, was the most effective for eliciting complex, epileptiform activity after benzyl-penicillin injection. The most complex epileptiform activity (spikes, and high frequency activity) was then repeated reliably in nine animals, utilizing 14 swine total. There were 20.1 +/- 10.8 [95% confidence interval (CI) 11.8-28.4] epileptiform events with > 3.5 Hz activity occurring per animal. Average duration of each event was 46.3 +/- 15.6 (95% CI 44.0-48.6) s, ranging from 20-100 s. In conclusion, the acute swine model of focal cortical epilepsy surgery provides an animal model that mimics human surgical conditions with a large brain and gyrated cortex, and is relatively inexpensive among animal models. Therefore, we feel this model provides a valuable, reliable, and novel platform for translational studies of implantable hardware for intracranial monitoring. PMID- 21627649 TI - Understanding the differences in prevalence of epilepsy in tropical regions. AB - Epilepsy is a frequent chronic neurologic disorder that affects nearly 70 million people worldwide. The majority of people with epilepsy live in developing countries, where epilepsy remains a major public health problem. Wide prevalence differences exist among various populations across sub-Saharan Africa, Latin America, and Asia. In particular, prevalence is lower in Southeast Asia than in sub-Saharan Africa and Latin America. Methodologic problems alone do not seem to explain these differences shown in recent review papers. The distribution of numerous risk or etiologic factors such as infectious diseases with neurologic sequel, head injuries, or genetic factors could explain these differences. Stigmatization of people with epilepsy could lead to underestimating the prevalence of epilepsy, even in well-conducted studies. It is important to standardize the process of epidemiologic monitoring of epilepsy in order to improve the reliability in data comparison. Understanding the reasons for these differences is a crucial issue for eventually raising new hypotheses or prevention strategies. PMID- 21627650 TI - Angiotensin II mediates urotensin II expression by hypoxia in cultured cardiac fibroblast. AB - BACKGROUND: Urotensin II plays a role in myocardial remodelling. Cardiac fibroblasts play a critical role in the development of cardiac fibrosis. The effect of hypoxia on urotensin II expression in cardiac fibroblasts is poorly understood. We sought to investigate the regulation of urotensin II by hypoxia in cardiac fibroblasts and the effect of angiotensin II in the interaction with urotensin II. METHODS AND RESULTS: Rat cardiac fibroblasts were cultured in hypoxic chamber. Hypoxia significantly increased urotensin II expression and reactive oxygen species (ROS) production in cultured cardiac fibroblasts. Hypoxia induced increase in urotensin II protein and ROS was significantly attenuated after the addition of SP600125, JNK siRNA or N-acetylcysteine before hypoxia treatment. The phosphorylated JNK protein was induced by hypoxia and was abolished by pretreatment with SP600125, losartan (an angiotensin II receptor antagonist) or N-acetylcysteine. The increased urotensin II expression by exogenous addition of angiotensin II was similar to that by hypoxia. Addition of losartan and angiotensin II antibody before hypoxia almost completely inhibited the increase in urotensin II induced by hypoxia. Hypoxia significantly increased the secretion of angiotensin II from cardiac fibroblasts and increased the collagen I protein expression. Hypoxia significantly increased the urotensin II promoter activity by 4.3-fold as compared to normoxic control. Urotensin II siRNA almost completely attenuated the collagen I protein expression induced by hypoxia. CONCLUSIONS: Hypoxia-induced urotensin II expression in cardiac fibroblast is mediated by angiotensin II and through ROS and JNK pathway. Urotensin II is a mediator of angiotensin II-induced cardiac fibrosis under hypoxia. PMID- 21627651 TI - Lactobacillus gasseri OLL2809 and its RNA suppress proliferation of CD4(+) T cells through a MyD88-dependent signalling pathway. AB - Recent studies have shown that probiotics are beneficial in prevention and improvement of inflammatory diseases. Accumulating evidence indicates that probiotics can modulate immune cell responses, although the specific molecular mechanism by which probiotics work remains elusive. Because T cells express receptors for microbial components, we examined whether the probiotic strain Lactobacillus gasseri OLL2809 (LG2809) and its components regulate murine CD4(+) T-cell activation. LG2809, as well as two other Lactobacillus strains, inhibited proliferation of CD4(+) T cells; LG2809 had the strongest suppressive activity among them. RNA isolated from LG2809 was also shown to have suppressive activity. We observed this suppressive effect in the culture of CD4(+) T cells stimulated with anti-CD3/CD28 treatment, suggesting a direct effect on CD4(+) T cells. In contrast, the suppressive effect was not observed for CD4(+) T cells from myeloid differentiation primary response gene 88 (MyD88) protein-deficient mice, and was abrogated in the presence of an anti-oxidant reagent, N-acetyl-cysteine (NAC). These results demonstrate that the suppressive effect of LG2809 and its RNA occurred through a MyD88-dependent signalling pathway and suggest involvement of a reactive oxygen species-dependent mechanism. LG2809 RNA injected subcutaneously suppressed delayed-type-hypersensitivity response in DO11.10 mice, and the suppression was abrogated by treatment with NAC. Collectively, these results suggest that suppression of T-cell proliferation by RNA may be one of the mechanisms when a probiotic bacterial strain exerts suppressive effects on inflammatory responses. PMID- 21627652 TI - Complexity of dendritic cell subsets and their function in the host immune system. AB - Dendritic cells (DCs) are professional antigen-presenting cells that are critical for induction of adaptive immunity and tolerance. Traditionally DCs have been divided into two discrete subtypes, which comprise conventional and non conventional DCs. They are distributed across various organs in the body and comprise a heterogeneous population, which has been shown to display differences in terms of surface marker expression, function and origins. Recent studies have shed new light on the process of DC differentiation and distribution of DC subtypes in various organs. Although monocytes, macrophages and DCs share a common macrophage-DC progenitor, a common DC progenitor population has been identified that exclusively gives rise to DCs and not monocytes or macrophages. In this review, we discuss the recent advances in our understanding of DC differentiation and subtypes and provide a comprehensive overview of various DC subtypes with emphasis on their function and origins. Furthermore, in light of recent developments in the field of DC biology, we classify DCs based on the precursor populations from which the various DC subsets originate. We classify DCs derived from common DC progenitor and pre-DC populations as conventional DCs, which includes both migratory and lymphoid-resident DC subsets and classify monocyte-derived DCs and plasmacytoid DCs as non-conventional DCs. PMID- 21627653 TI - The genetic etiology of inhibitory control and behavior problems at 24 months of age. AB - BACKGROUND: To investigate links between inhibitory control (IC) and behavior problems in early childhood, as well as genetic and environmental covariances between these two constructs. METHODS: Parent and laboratory ratings of IC and parent ratings of externalizing and attention deficit hyperactivity disorder behaviors were administered at 24 months of age on a sample of 291 same-sex twin pairs (131 monozygotic, 160 dizygotic). RESULTS: There were significant phenotypic associations between both IC assessments and the two areas of behavioral maladjustment (correlations ranged from -.13 to -.57). Multivariate analyses revealed that phenotypic covariance between IC and behavior problems could be substantially explained by common genetic influences (genetic correlations ranged from -.30 to -.74). Parent ratings of IC showed higher phenotypic and genetic correlations with behavior problems than lab ratings of IC. CONCLUSIONS: This study is the first to examine the etiology of the covariance between IC and related behavioral difficulties in toddlerhood. Findings suggest that low levels of IC can be considered a genetic risk factor for the development of early emerging behavior problems. PMID- 21627654 TI - Do clinicians really need to understand statistics? PMID- 21627655 TI - Outcome in 55 dogs with pulmonic stenosis that did not undergo balloon valvuloplasty or surgery. AB - OBJECTIVE: To determine the outcome, independent predictors of cardiac death, and the Doppler-derived pressure gradient cut-off for predicting cardiac death in dogs with pulmonic stenosis, with or without tricuspid regurgitation, that do not undergo balloon valvuloplasty or valve surgery. METHODS: Review of medical records of two UK referral centres between July 1997 and October 2008 for all cases of pulmonic stenosis that had no balloon valvuloplasty or valve surgery. Inclusion criteria included a diagnosis of pulmonic stenosis; spectral Doppler pulmonic velocity greater than 1.6 m/s; characteristic valve leaflet morphological abnormalities. Exclusion criteria included concurrent significant cardiac defects, including tricuspid dysplasia. Dogs with tricuspid regurgitation were included. Dogs were classified according to Doppler-derived pressure gradients into mild, moderate or severe pulmonic stenosis categories. RESULTS: Presence of tricuspid regurgitation and severe stenosis were independent predictors of cardiac death. A pulmonic pressure gradient of more than 60 mmHg was associated with 86% sensitivity, and 71% specificity of predicting cardiac death. CLINICAL SIGNIFICANCE: There is an increased probability of cardiac death in those cases which have a pulmonary pressure gradient greater than 60 mmHg and tricuspid regurgitation, though the effect of severity of tricuspid regurgitation on outcome was not measurable because of small sample sizes. These animals might benefit from intervention. PMID- 21627656 TI - Laparoscopic ovariectomy in dogs: a comparison of the LigaSureTM and the SonoSurgTM systems. AB - OBJECTIVES: The objective of this study was to evaluate the surgical times for removal of ovaries, and the frequency of intra-operative complications with two different instruments used for canine laparoscopic ovariectomy. METHODS: A randomised prospective clinical study. Laparoscopic ovariectomy was performed under general anaesthesia on 10 healthy female dogs admitted for elective ovariectomy. Each ovary was randomly assigned to removal by use of either a SonoSurgTM ultrasonic surgical device (Olympus), or a LigaSureTM vessel-sealing system (Valleylab/Covidien). Surgical time for removal of each ovary was measured and the frequency of intra-operative complications was recorded. A follow-up was made by interview with the owners, 2.5 years after surgery. RESULTS: The two techniques did not statistically differ with regard to surgical time, or frequency of complications. All owners were very satisfied with the procedure. CLINICAL RELEVANCE: Both the SonoSurgTM and the LigaSureTM devices appear to be effective, safe and easy to use when performing laparoscopic ovariectomy in dogs. PMID- 21627657 TI - Suspected contact scrotal dermatitis in the dog: a retrospective study of 13 cases (1987 to 2003). AB - OBJECTIVES: The aim of this study was to assess the clinical and aetiological aspects of contact dermatitis of the scrotum in dogs. METHODS: The medical records of 13 dogs with suspected scrotal contact dermatitis were retrospectively included in the study. For each case, historical findings, results of physical and dermatological examination and of avoidance tests as well as follow-up were included. Results of re-exposure tests and patch tests were also included when available. RESULTS: Avoidance tests to various suspected substances such as floor detergents, bleach, cement, laundry detergent and plastic fabric were performed and led to clinical improvement in all patients. Provocative re-exposure was performed in six cases, with five positive responses. Patch tests were undertaken in three of the five re-exposed cases, with two responding positively. CLINICAL SIGNIFICANCE: Scrotal skin is thin and with few hair follicles, and is therefore very susceptible to contact dermatitis. A range of substances can induce contact dermatitis. The diagnosis of contact dermatitis is made by results of avoidance and/or provocation tests and/or patch tests. Once an aetiological agent has been identified, treatment is straightforward consisting of avoidance of this agent. PMID- 21627658 TI - Determination of haematological reference intervals in healthy adult greyhounds. AB - OBJECTIVE: The purpose of this study was to establish breed-specific reference intervals for haematological measurands in non-racing greyhounds. Suitability of the data for partitioning according to sex was also examined. METHODS: Haematological data were collected from 304 healthy non-racing greyhounds and analysed using non-parametric methods. Results were compared with non-breed specific canine reference intervals and also with greyhound reference intervals obtained by other investigators. RESULTS: Compared with non-breed-specific reference intervals, the results showed comparable mean and upper limit and higher lower limit for erythrocyte count; higher values for haemoglobin, haematocrit and mean corpuscular volume; and lower values for total leucocyte count and absolute concentration of neutrophils, lymphocytes, monocytes, eosinophils and platelets. Partitioning according to sex was recommended by the statistical analysis for all analytes except haematocrit and total leucocyte count. CLINICAL SIGNIFICANCE: In this study the reference intervals were derived from a large sample size. The results are in general agreement with previous reports, although higher values for low reference limits have been noted for the erythroid parameters, and lower values for upper reference limits have been observed for the total and different leucocyte counts. Breed-specific reference intervals provide a useful clinical tool for haematological evaluations. PMID- 21627659 TI - Therapeutic antibiotic use patterns in dogs: observations from a veterinary teaching hospital. AB - OBJECTIVES: To describe therapeutic antibiotic use patterns in dogs at a small animal teaching hospital. METHODS: A retrospective case analysis of randomly sampled antibiotic prescriptions in dogs from May 20, 2008 to May 20, 2009, deemed to be for therapeutic use, was performed. Records were reviewed to determine if there was documentation of confirmed, suspected or no evidence of infection. The five most frequently prescribed antibiotics were identified and analysed for their distribution in these categories. RESULTS: In 17% of therapeutic antibiotic prescriptions there was confirmed infection, in 45% suspected infection, and in 38% there was no documented evidence of infection. Amoxicillin-clavulanate was the most frequently prescribed antibiotic, followed by cefazolin/cephalexin, enrofloxacin, ampicillin/amoxicillin and doxycycline. Doxycycline was the most frequently prescribed with no documented evidence of infection, and amoxicillin-clavulanate was the most frequently prescribed with either confirmed or suspected evidence of infection. DISCUSSION: Clinicians use a variety of tools when deciding whether or not to prescribe an antibiotic and which antibiotic to use. As in human medicine, there is likely overuse and inappropriate use of antibiotics in veterinary medicine. Veterinarians should engage in discussions regarding clinically applicable guidelines for appropriate antibiotic use. PMID- 21627660 TI - Dermoid sinus and spina bifida in three dogs and a cat. AB - This case series describes four cases of concomitant dermoid sinus and spina bifida in a Chinese crested dog, two Swedish vallhunds and a Burmese cat. The diagnosis was confirmed by computed tomography, computed tomography fistulography and pathology. Two dogs that showed abnormalities during neurological examination underwent magnetic resonance imaging. One of them had imaging findings interpreted as syringohydromyelia. All the dogs underwent surgery and the outcome was considered good in all the cases. Based on clinical and pathological examination as well as diagnostic imaging findings, two of the dermoid sinuses were classified as type IV, and two of them showed a previously unreported type of dermoid sinus. This new type is suggested here as type VI. PMID- 21627661 TI - Intracranial anaplastic astrocytoma in a 19-week-old boxer dog. AB - A 19-week-old, entire female boxer dog was presented for evaluation of chronic, progressive multi-focal neurological signs. Magnetic resonance imaging suggested a large intra-axial mass in the brainstem. Euthanasia was performed and an anaplastic astrocytoma was found at post-mortem examination. This case reports an unusual naturally occurring brain tumour in a young dog. PMID- 21627662 TI - Image-guided transnasal cryoablation of a recurrent nasal adenocarcinoma in a dog. AB - An eight-year-old female spayed Airedale terrier with rapid recurrence of a nasal adenocarcinoma following image-guided intensity-modulated radiation therapy was treated with transnasal, image-guided cryotherapy. Ice ball size and location were monitored real-time with computed tomography-fluoroscopy to verify that the entire tumour was enveloped in ice. Serial computed tomography scans demonstrated reduction in and subsequent resolution of the primary tumour volume corresponding visually with the ice ball imaged during the ablation procedure. Re-imaging demonstrated focallysis of the cribriform plate following ablation that spontaneously resolved by 13 months. While mild chronic nasal discharge developed following cryoablation, no other clinical signs of local nasal neoplasia were present. Twenty-one months after nasal tumour cryoablation the dog was euthanased as a result of acute haemoabdomen. Image-guided cryotherapy may warrant further investigation for the management of focal residual or recurrent tumours in dogs, especially in regions where critical structures preclude surgical intervention. PMID- 21627664 TI - Xylem traits mediate a trade-off between resistance to freeze-thaw-induced embolism and photosynthetic capacity in overwintering evergreens. AB - Hydraulic traits were studied in temperate, woody evergreens in a high-elevation heath community to test for trade-offs between the delivery of water to canopies at rates sufficient to sustain photosynthesis and protection against disruption to vascular transport caused by freeze-thaw-induced embolism. Freeze-thaw-induced loss in hydraulic conductivity was studied in relation to xylem anatomy, leaf- and sapwood-specific hydraulic conductivity and gas exchange characteristics of leaves. We found evidence that a trade-off between xylem transport capacity and safety from freeze-thaw-induced embolism affects photosynthetic activity in overwintering evergreens. The mean hydraulically weighted xylem vessel diameter and sapwood-specific conductivity correlated with susceptibility to freeze-thaw induced embolism. There was also a strong correlation of hydraulic supply and demand across species; interspecific differences in stomatal conductance and CO(2) assimilation rates were correlated linearly with sapwood- and leaf-specific hydraulic conductivity. Xylem vessel anatomy mediated an apparent trade-off between resistance to freeze-thaw-induced embolism and hydraulic and photosynthetic capacity during the winter. These results point to a new role for xylem functional traits in determining the degree to which species can maintain photosynthetic carbon gain despite freezing events and cold winter temperatures. PMID- 21627666 TI - The role of heterotrophic carbon acquisition by the hemiparasitic plant Rhinanthus alectorolophus in seedling establishment in natural communities: a physiological perspective. AB - * Heterotrophic acquisition of substantial amounts of organic carbon by hemiparasitic plants was clearly demonstrated by numerous studies. Many hemiparasites are, however, also limited by competition for light preventing the establishment of their populations on highly productive sites. * In a growth chamber experiment, we investigated the effects of competition for light, simulated by shading, on growth and heterotrophic carbon acquisition by the hemiparasite Rhinanthus alectorolophus attached to C(3) and C(4) hosts using analyses of biomass production and stable isotopes of carbon. * Shading had a detrimental effect on biomass production and vertical growth of the hemiparasites shaded from when they were seedlings, while shading imposed later caused only a moderate decrease of biomass production and had no effect on the height. Moreover, shading increased the proportion of host-derived carbon in hemiparasite biomass (up to 50% in shaded seedlings). * These results demonstrate that host derived carbon can play a crucial role in carbon budget of hemiparasites, especially if they grow in a productive environment with intense competition for light. The heterotrophic carbon acquisition can allow hemiparasite establishment in communities of moderate productivity, helping well-attached hemiparasites to escape from the critical seedling stage. PMID- 21627667 TI - Photochemical reflectance index (PRI) and remote sensing of plant CO2 uptake. PMID- 21627665 TI - Invasion potential and host shifts of Australian and African ectomycorrhizal fungi in mixed eucalypt plantations. AB - * Transportation of forestry materials results in unintended co-introduction of nonnative species that may cause enormous ecological or economic damage. While the invasion ecology of plants and animals is relatively well-known, that of microorganisms, except aboveground pathogens, remains poorly understood. * This work addresses host shifts and invasion potential of root symbiotic ectomycorrhizal fungi that were co-introduced with Australian eucalypts and planted in clear-cut miombo woodlands in Zambia, south-central Africa. * By use of rDNA and plastid intron sequence analysis for identification and phylogenetic techniques for inferring fungal origin, we demonstrated that host shifts were uncommon in the Australian fungi, but frequent in the African fungi, especially in mixed plantations where roots of different trees intermingle. * There was evidence for naturalization, but not for invasion by Australian ectomycorrhizal fungi. Nevertheless, the fungi introduced may pose an invasion risk along with further adaptation to local soil environment and host trees. Inoculation of eucalypts with native edible fungi may ameliorate the potential invasion risks of introduced fungi and provide an alternative source of nutrition. PMID- 21627669 TI - Congenital narrowing of a pulmonary vein: slit-like pulmonary vein ostium. AB - We report the case of a 60-year-old female with a history of refractory paroxysmal atrial fibrillation. Preablation contrast enhanced pulmonary vein computed tomography (CT) scan demonstrated a slit-like narrowing of the left inferior pulmonary vein ostium. The narrowing measured approximately 3 mm, with poststenotic dilation. The patient had no prior history of ablation. The patient subsequently underwent segmental antral isolation of all four pulmonary veins and cavo-tricuspid isthmus ablation with bidirectional block. The diagnosis of preexisting congenital pulmonary vein stenosis had an impact on the type of ablation procedure performed (antral rather than ostial) and will affect the interpretation of postablation CT scans. PMID- 21627670 TI - Exploring the composition and diversity of microbial communities at the Jan Mayen hydrothermal vent field using RNA and DNA. AB - DNA sequencing technology has proven very valuable for analysing the microbiota of poorly accessible ecosystems such as hydrothermal vents. Using a combination of amplicon and shotgun sequencing of small-subunit rRNA and its gene, we examined the composition and diversity of microbial communities from the recently discovered Jan Mayen vent field, located on Mohn's Ridge in the Norwegian Greenland Sea. The communities were dominated by the epsilonproteobacterial genera Sulfurimonas and Sulfurovum. These are mesophiles involved in sulphur metabolism and typically found in vent fluid mixing zones. Composition and diversity predictions differed systematically between extracted DNA and RNA samples as well as between amplicon and shotgun sequencing. These differences were more substantial than those between two biological replicates. Amplicon vs. shotgun sequencing differences could be explained to a large extent by bias introduced during PCR, caused by preferential primer-template annealing, while DNA vs. RNA differences were thought to be caused by differences between the activity levels of taxa. Further, predicted diversity from RNA samples was consistently lower than that from DNA. In summary, this study illustrates how different methods can provide complementary ecological insights. PMID- 21627671 TI - Soil factors exhibit greater influence than bacterial inoculation on alfalfa growth and nitrogen fixation. AB - In order to study the effects of soil factors and bacterial inoculation on alfalfa (Medicago sativa), plants were inoculated with Ensifer meliloti L33 and Azospirillum brasilense Sp7 in pot experiments using two different soils separately as well as in a mixture. One soil was contaminated with chemical waste products; the other was an arable soil. Soil factors, including the availability of macro- and micronutrients as well as carbon and nitrogen contents, were found to exhibit a much greater influence on the growth of alfalfa than any of the inoculations. In contaminated soil, the shoot and root growth of alfalfa was decreased and nodules were diminished and ineffective. Bacterial inoculations did not significantly improve this hostile growth environment. However, in a mixture (44% arable, 22% contaminated soil, 34% vermiculite), growth conditions for alfalfa were improved so that shoot dry weight and nodule numbers increased up to 100- and 20-fold, respectively, compared with the contaminated soil. For the strain L33, its persistence in the rhizosphere was correlated to the presence of its host plant, but its dynamics were influenced by competition with indigenous rhizobia. The strain Sp7, once provided with a suitable soil, was not dependent on the plant's rhizosphere, but it enhanced the performance of L33 and native rhizobia. PMID- 21627672 TI - Estimating the national public health burden associated with exposure to ambient PM2.5 and ozone. AB - Ground-level ozone (O(3)) and fine particulate matter (PM(2.5)) are associated with increased risk of mortality. We quantify the burden of modeled 2005 concentrations of O(3) and PM(2.5) on health in the United States. We use the photochemical Community Multiscale Air Quality (CMAQ) model in conjunction with ambient monitored data to create fused surfaces of summer season average 8-hour ozone and annual mean PM(2.5) levels at a 12 km grid resolution across the continental United States. Employing spatially resolved demographic and concentration data, we assess the spatial and age distribution of air-pollution related mortality and morbidity. For both PM(2.5) and O(3) we also estimate: the percentage of total deaths due to each pollutant; the reduction in life years and life expectancy; and the deaths avoided according to hypothetical air quality improvements. Using PM(2.5) and O(3) mortality risk coefficients drawn from the long-term American Cancer Society (ACS) cohort study and National Mortality and Morbidity Air Pollution Study (NMMAPS), respectively, we estimate 130,000 PM(2.5) -related deaths and 4,700 ozone-related deaths to result from 2005 air quality levels. Among populations aged 65-99, we estimate nearly 1.1 million life years lost from PM(2.5) exposure and approximately 36,000 life years lost from ozone exposure. Among the 10 most populous counties, the percentage of deaths attributable to PM(2.5) and ozone ranges from 3.5% in San Jose to 10% in Los Angeles. These results show that despite significant improvements in air quality in recent decades, recent levels of PM(2.5) and ozone still pose a nontrivial risk to public health. PMID- 21627673 TI - Health literacy -- a heterogeneous phenomenon: a literature review. AB - BACKGROUND AND AIM: A growing responsibility on the part of individuals to make decisions in health issues implies the need of access to health information and personal skills to comprehend the information. Health literacy comprises skills in obtaining, understanding and acting on information about health issues in ways that promote and maintain health. A lack of health literacy may have effects at both the individual and societal levels. There are thus reasons for health care professionals to gain a comprehensive understanding of health literacy. The aim of this review was to explore how health literacy is described in the scientific literature and to give a synthesis of its different meanings. METHODS: The review was based on approximately 200 scientific articles published 2000-2008. The analysis process was inspired by the methods of narrative literature review. FINDINGS AND CONCLUSIONS: Two different approaches to health literacy became visible, one in which health literacy is expressed as a polarized phenomenon, focusing on the extremes of low and high health literacy. The definitions of health literacy in this approach are characterized by a functional understanding, pointing out certain basic skills needed to understand health information. The other approach represents a complex understanding of health literacy, acknowledging a broadness of skills in interaction with the social and cultural contexts, which means that an individual's health literacy may fluctuate from one day to another according to the context. The complex approach stresses the interactive and critical skills needed to use information or knowledge as a basis for appropriate health decisions. We conclude that health literacy is a heterogeneous phenomenon that has significance for both the individual and society. Future research will aim at the development of assessments that capture the broadness of skills and agents characteristic for health literacy as a complex phenomenon. PMID- 21627674 TI - A novel deletion of the MEN1 gene in a large family of multiple endocrine neoplasia type 1 (MEN1) with aggressive phenotype. AB - CONTEXT: The MEN1 syndrome is associated with parathyroid, pancreatic and pituitary tumours and is caused by mutations in the MEN1 gene. In general, there is no genotype-phenotype correlation. OBJECTIVES: To characterize a large family with MEN1 with aggressive tumour behaviour: malignant pancreatic endocrine tumours were present in five affected subjects and were the presenting features in three subjects. DESIGN: The coding region of MEN1 was sequenced. Gene copy number analysis was performed by multiplex ligation-dependent probe amplification (MLPA) and array comparative genomic hybridization (aCGH). Loss of heterozygosity (LOH) in tumour tissue was studied by microsatellite analysis. Insulin-like growth factor II (IGF-II) and CDKN1C/p57KIP2 expression were investigated by immunohistochemistry. RESULTS: Mutation screening by conventional PCR sequence analysis of patients' peripheral blood DNA did not reveal any mutation in the MEN1 or CDKN1B gene. Gene copy number analysis by MLPA and aCGH demonstrated a novel monoallelic deletion of 5 kb genomic DNA involving the MEN1 promoter and exons 1 and 2. LOH analysis indicated somatic deletion of maternal chromosome 11, including MEN1 locus (11q13) and 11p15 imprinting control regions (ICR). Methylation analysis of ICR demonstrated ICR1 hypermethylation and ICR2 hypomethylation in the tumour specimens. ICR1 and ICR2 control the expression of IGF-2 and CDKN1C/p57KIP2, respectively. Immunohistochemistry showed that expression of paternally expressed IGF-2 was up-regulated and the maternally expressed CDKN1C/p57KIP2 was lost in the pancreatic endocrine tumours. CONCLUSIONS: Gene copy number analysis by MLPA should be considered in patients with negative conventional mutation screening. Although large MEN1 deletion causes MEN1, disruption of imprinted CDKN1C/p57KIP2 and IGF-2 gene expression may contribute to tumour progression and aggressive phenotype. PMID- 21627675 TI - Retention of questionable and hopeless teeth in compliant patients treated for aggressive periodontitis. AB - AIM: The aim of this study was to determine the survival rates of questionable and hopeless teeth in patients with aggressive periodontitis (AgP) and chronic periodontitis (CP) during 15 years of supportive periodontal therapy (SPT). MATERIALS AND METHODS: Thirty-four AgP and 34 CP patients (SPT>=10 years) with bone loss of >=50% at >=2 teeth were consecutively recruited. Bone loss was measured on digitized radiographs and teeth were categorized as "questionable" (>=50 to <70% bone loss) or as "hopeless" (>=70%). Progression in pocket probing depths (PPD) during SPT, tooth loss and reasons for extraction were analysed. RESULTS: In AgP patients, 262 teeth were considered as questionable and 63 as hopeless (CP: 149/51). During active periodontal therapy, 25 questionable and 26 hopeless teeth were extracted (CP: 12/16). During 15.3 +/- 4.1 years of SPT of AgP 28 questionable and 15 hopeless teeth were removed (CP: 28/12). The mean tooth loss per patient during SPT in total was 0.14 (AgP) and 0.16 (CP) teeth/year. There were no significant differences in tooth loss or longitudinal progression of PPD between AgP and CP patients. CONCLUSIONS: In patients with AgP, 88.2% (209 of 237) of questionable and 59.5% (22 of 37) of hopeless teeth survived 15 years during regular SPT in a dental school department. PMID- 21627676 TI - Risk of meningioma among users of high doses of cyproterone acetate as compared with the general population: evidence from a population-based cohort study. AB - AIM: Information from the spontaneous reporting system raised the hypothesis of an increased risk of meningioma in patients treated with high doses of cyproterone acetate (CPA). The objective of this study was to test the hypothesis of an increased risk of meningioma among users of high dose CPA as compared with non-users in a medical records computerized database. METHODS: A retrospective cohort study was performed in a Spanish primary care database (BIFAP). Meningioma incidence rates were compared in patients exposed to high dose CPA (users) with those non-exposed and with those exposed to low dose CPA. Poisson regression analysis was used to estimate the incidence rate ratios after adjusting for age and gender. RESULTS: Among 2474 users of high dose cyproterone (6663 person years) four meningioma cases were identified, resulting in an incidence rate (IR) of 60.0 (95% CI 16.4, 153.7) per 100,000 person-years, which was significantly higher than that observed among the non-users (IR 6.6; 95% CI 6.0, 7.3) and among women users of low dose cyproterone (IR 0.0, 95% CI upper limit 5.5). After adjusting for age and gender, patients exposed to high dose CPA showed an increased risk of meningioma of 11.4 (95% CI 4.3, 30.8) as compared with non users. CONCLUSIONS: The results of this study support the hypothesis that the exposure to high dose CPA increases the risk of meningioma. PMID- 21627677 TI - Electronic prescribing increases uptake of clinical pharmacologists' recommendations in the hospital setting. AB - AIMS: To determine whether electronic prescribing facilitates the uptake of clinical pharmacologists' recommendations for improving drug safety in medical inpatients. METHODS: Electronic case records and prescription charts (either electronic or paper) of 502 patients hospitalized on medical wards in a large Swiss teaching hospital between January 2009 and January 2010 were studied by four junior and four senior clinical pharmacologists. Drug-related problems were identified and interventions proposed. The implementation and time delays of these proposed interventions were compared between the patients for whom paper drug charts were used and the patients for whom electronic drug charts were used. RESULTS: One hundred and fifty-eight drug-related problems in 109 hospital admissions were identified and 145 recommendations were made, of which 51% were implemented. Admissions with an electronic prescription chart (n= 90) were found to have 2.74 times higher odds for implementation of the change than those with a paper prescription chart (n= 53) (95% confidence interval 1.2, 6.3, P= 0.018, adjusted for any dependency introduced by patient, ward or clinical team; follow up for two cases missing). The time delay between recommendations being made and their implementation (if any) was minimal (median 1 day) and did not differ between the two groups. CONCLUSIONS: Electronic prescribing in this hospital setting was associated with increased implementation of clinical pharmacologists' recommendations for improving drug safety when compared with handwritten prescribing on paper. PMID- 21627678 TI - A discussion of approaches to transforming care: contemporary strategies to improve patient safety. AB - AIM: This article presents a discussion of three contemporary approaches to transforming care: Transforming Care at the Bedside, Releasing Time to Care: the Productive Ward and the work of the Studer Group((r)). BACKGROUND: International studies of adverse events in hospitals have highlighted the need to focus on patient safety. The case for transformational change was identified and recently several approaches have been developed to effect this change. Despite limited evaluation, these approaches have spread and have been adopted outside their country of origin and contextual settings. DATA SOURCES: Medline and CINAHL databases were searched for the years 1999-2009. Search terms included derivatives of 'transformation' combined with 'care', 'nursing', 'patient safety', 'Transforming Care at the Bedside', 'the Productive Ward' and 'Studer Group'. DISCUSSION: A comparison of the three approaches revealed similarities including: the foci of the approaches; interventions employed; and the outcomes measured. Key differences identified are the implementation models used, spread strategies and sustainability of the approaches. The approaches appear to be complementary and a hybrid of the approaches such as a blend of a top-down and bottom-up leadership strategy may offer more sustainable behavioural change. IMPLICATIONS FOR NURSING: These approaches transform the way nurses do their work, how they work with others and how they view the care they provide to promote patient safety. CONCLUSION: All the approaches involve the implementation of multiple interventions occurring simultaneously to affect improvements in patient safety. The approaches are complementary and a hybrid approach may offer more sustainable outcomes. PMID- 21627679 TI - Older people's views in relation to risk of falling and need for intervention: a meta-ethnography. AB - AIM: This paper is a report of a meta-ethnography of qualitative studies of older peoples' views on risk of falling and need for intervention. BACKGROUND: Falls and falls-related injuries in older people are worldwide problems. A conceptual understanding of older people's views about falls risk and need for intervention is useful for understanding factors likely to impact on acceptance of risk and recommended interventions. DATA SOURCES: Seven electronic databases were searched 1999-2009. Reference lists of included articles were screened for eligible papers. REVIEW METHODS: Assessment of quality was carried out. Themes and concepts were extracted using a meta-ethnographic approach to compare similarities and differences across the retrieved studies. A line of argument was developed to produce an explanatory framework of the extracted themes and concepts. RESULTS: Eleven relevant qualitative research articles of reasonable quality were identified. Six key concepts were identified: beyond personal control; rationalizing; salience; life-change and identity; taking control and self-management. A line of argument synthesis describes how older people approach self-appraisal of falls risk and intervention need, and how they cope and adapt to falls risk and intervention need. CONCLUSION: In response to having an elevated risk status and perceived associations with frailty and impact on an independent life-style, some prefer to adapt to this reality by taking control and implementing self-management strategies. Healthcare professionals should take into account beliefs about risk and negotiate choices for intervention, recognizing that some individuals prefer to drive the decision-making process to preserve identity as a competent and independent person. PMID- 21627680 TI - The intention to continue nursing: work variables affecting three nurse generations in Australia. AB - AIMS: The aims of the study were to examine how seven variables impacted upon the intention of hospital nurses to continue working as nurses and to investigate whether there are generational differences in these impacts. BACKGROUND: There is a critical shortage of trained nurses working as nurses in Australia, as in many other Organisation for Economic Co-operation and Development member countries. The retention of nurses has been examined from the traditional management perspectives; however, this paper presents a different approach (Meaning of Working theory). METHODS: A self-report survey of 900 nurses employed across four states of Australia was completed in 2008. The sample was hospital nurses in Australia from three generational cohorts - Baby Boomers (born in Australia between 1946 and 1964), Generation X (1965-1979) and Generation Y (1980-2000). RESULTS: Six variables were found to influence the combined nurses' intentions to continue working as nurses: work-family conflict, perceptions of autonomy, attachment to work, importance of working to the individual, supervisor subordinate relationship and interpersonal relationships at work. There were differences in the variables affecting the three generations, but attachment to work was the only common variable across all generations, affecting GenYs the strongest. CONCLUSION: The shortage of nurses is conceptualized differently in this paper to assist in finding solutions. However, the results varied for the three generations, suggesting the need to tailor different retention strategies to each age group. Implications for management and policy planning are discussed. PMID- 21627681 TI - The effect of sexual self-concept on sexual health behavioural intentions: a test of moderating mechanisms in early adolescent girls. AB - AIM: The purpose of this study was to examine whether normative beliefs would act as a moderator of the main relationship between sexual self-concept and sexual heath behavioural intentions. BACKGROUND: Sexual self-concept has been defined as an individual's evaluation of his or her own sexual feelings and actions. Research has shown that sexually related behaviours are associated with sexual self-concept and perceived parental/peer approval of sexual behaviour. METHODS: We conducted cross-sectional research, using face-to-face interviews, to assess sexual self-concept, normative beliefs and sexual health behavioural intention. This study was conducted with 534 adolescent female participants, aged 12-15 years, in Taiwan. Data was collected between September and December 2009. We conducted structural equation modelling to examine the proposed conceptual model. FINDINGS: We found that individuals with a high score on sexual self-concept (i.e. who had stronger erotic feelings) were less likely to believe that they would behave in ways that would protect their sexual health (i.e. low behavioural intention, beta = -0.44, t = -8.43, P < 0.001). The hypothesized moderating role of normative beliefs was also supported. Those with a greater tendency to believe that their parents/peers approved of sex (i.e. normative beliefs) were more likely to have a positive sexual self-concept (beta = 0.57, P < 0.001), and were less likely to protect their sexual health (beta = -0.19, P < 0.01). The findings showed that, together, normative beliefs and sexual self-concept accounted for 24.9% of the variance in girls' sexual health behavioural intentions. CONCLUSIONS: We recommend that interventions that target sexual health behavioural intentions for this population would be more effective if such interventions targeted normative beliefs and helped adolescent females to clarify their sexual self-concept. PMID- 21627682 TI - Relieving existential suffering through palliative sedation: discussion of an uneasy practice. AB - AIM: This article presents a discussion of the use of palliative sedation in response to intractable (not responsive to treatment) existential suffering. BACKGROUND: Patients suffering from a terminal illness are often faced with severe symptoms at the end of life. Although palliative sedation is sometimes used when no other options are effective in relieving unbearable pain or suffering, its use in response to intractable existential suffering in terminal illness remains controversial. DATA SOURCES: A literature search was conducted for published articles addressing the use of palliative sedation between 1996 and 2009 using established databases. DISCUSSION: Palliative sedation remains an uneasy practice. The debates have centred on ethical issues surrounding decisions to use sedation and on separating the intent of palliative sedation (relief of intolerable symptoms) from the intent of euthanasia (hastening death). There is lack of consensus in defining existential suffering. Consequently, there is limited understanding of how decisions are being made when using palliative sedation to treat intractable existential suffering. CONCLUSIONS: Given the confusion and uncertainty about ethical and clinical justifications for palliative sedation in treating existential suffering, we argue that a better understanding of the controversies and decision-making process is needed. Greater understanding is required to prevent palliative sedation from becoming a substitute for intensive treatment of this kind of suffering. PMID- 21627683 TI - An international collaborative study comparing Swedish and Japanese nurses' reactions to elder abuse. AB - AIM: This paper reports an analysis of aggregated data from two national studies on Swedish community-based nurses' and Japanese Public Health Nurses' responses to hypothetical elder abuse cases. BACKGROUND: Elder abuse is an under-researched area despite being globally recognized as a serious and escalating problem. Yet research, adding needed socio-cultural perspectives to current knowledge has been limited. METHODS: Eighty-one community-based nurses in Sweden and 124 Public Health Nurses in Japan responded to a questionnaire based on three hypothetical elder abuse cases. Swedish and Japanese results (data collection 2006-2007) were combined and the aggregated data were analysed using manifest and qualitative content analyses. RESULTS: Nurses' response patterns in the aggregated data were similar across all three hypothetical cases and within themes Awareness, Assessment and Intervention. However, there were also noteworthy differences between Swedish and Japanese responses, e.g. Swedish responses were generally practical, action oriented and involved increased levels of suspicion and personal intervention to achieve increased control; whereas Japanese responses concerned better understanding that involved the family members and their situation, focusing on interventions grounded in collaboration. CONCLUSION: Despite cultural differences, responses of Swedish and Japanese nurses were very similar which points to a global 'humanness' of the problem of, and nurses' responses to, elder abuse. Results endorse the value of international collaborations that give information and inspiration to nursing colleagues across cultural boundaries. Results also give hope that global tools for elder abuse assessment and intervention can be developed. PMID- 21627684 TI - A toast to Sir Harold Himsworth. PMID- 21627685 TI - Experience of hypoglycaemia is associated with changes in beliefs about diabetes in patients with type 2 diabetes. AB - AIM: Hypoglycaemia may have a detrimental impact on quality of life for patients with Type 2 diabetes. There are few clinical studies exploring the impact of experiencing hypoglycaemia on beliefs about diabetes and health status. The aim of this study was to explore associations between experience of hypoglycaemia and changes in diabetes beliefs and self-reported health status in patients with non insulin-treated Type 2 diabetes using a blood glucose meter. METHODS: One-year prospective cohort analysis of 226 patients recruited to a randomized trial evaluating the impact of self-monitoring of blood glucose. Self-reported hypoglycaemia over 1 year was categorized into three groups: (1) no experience of hypoglycaemia; (2) blood glucose measurements < 4 mmol/l with no associated symptoms of hypoglycaemia (grade 1); and (3) symptomatic hypoglycaemia (grade 2 and 3). Measures of beliefs about diabetes (Revised Illness Perception Questionnaire) and health status (EuroQol-5D) were assessed at baseline and 1 year. Differences in mean changes over 1 year were explored with analyses of covariance. RESULTS: There was a significant increase in mean score in beliefs about personal control (1.14; 95%CI 0.14-2.14) among those experiencing grade 1 hypoglycaemia compared with those not experiencing hypoglycaemia. There were no significant differences in changes in health status between groups, with small overall changes that were inconsistent between groups. CONCLUSIONS: This study does not provide support for a long-term adverse impact on beliefs about diabetes or health status from the experience of mild symptomatic hypoglycaemia, in well controlled, non-insulin-treated patients with Type 2 diabetes using self monitoring of blood glucose. PMID- 21627686 TI - Evidence for a dose effect of renin-angiotensin system inhibition on progression of microalbuminuria in Type 2 diabetes: a meta-analysis. AB - AIMS: Renin-angiotensin inhibitors in Type 2 diabetes and microalbuminuria reduce renal and cardiovascular risk, but evidence supporting use of maximal tolerated dose is unclear. We aimed to determine the extent of renin-angiotensin inhibitor dose-dependent effects from randomized trials carried out in a clinical setting. METHODS: In a meta-analysis of randomized clinical trials, alternate doses of angiotensin receptor blockers or angiotensin converting enzyme inhibitors in patients with Type 2 diabetes and microalbuminuria were compared. MEDLINE, EMBASE and the Cochrane Register of Controlled Trials were searched from January 2006 to August 2010. Trials prior to January 2006 were identified from a prior systematic review. Identified outcomes were albumin excretion rate, progression and regression of albuminuria and adverse events. RESULTS: Four trials including 1051 patients compared doses of angiotensin receptor blockers. No trials compared doses of angiotensin converting enzyme inhibitor. The percentage decline in albumin excretion rate from baseline was greater with higher doses (18% higher, 95% CI 8-28%), the regression to normoalbuminuria was greater (OR 1.66, 95% CI 1.22-2.27), with less progression to macroalbuminuria (OR 0.62, CI 0.38-1.02). Adverse events were fewer with lower-dose angiotensin receptor blockers (OR 1.32, 95% CI 0.90-1.92). CONCLUSIONS: Higher-dose compared with lower-dose angiotensin receptor blockers in Type 2 diabetes with microalbuminuria are associated with significantly reduced albumin excretion rate and increased regression to normoalbuminuria. Adverse events are more frequent, but not significantly so. There is potential for trials to determine clinical cardiovascular and renal outcomes at differing doses. Our findings support current recommendations to titrate renin-angiotensin inhibitors to maximum dose whilst considering risk of adverse side effects with higher doses. PMID- 21627687 TI - A new model for 5-year risk of cardiovascular disease in Type 1 diabetes; from the Swedish National Diabetes Register (NDR). AB - AIMS: We assessed the association between risk factors and cardiovascular disease in an observational study of patients with Type 1 diabetes from the Swedish National Diabetes Register. METHODS: A derivation sample of 3661 patients, aged 30-65 years, 6.1% with previous cardiovascular disease, baseline 2002, and 197 cardiovascular disease events when followed for 5 years until 2007. A separate validation data set of 4484 patients, baseline 2003, 201 cardiovascular disease events when followed for 4 years. RESULTS: Adjusted hazard ratios at Cox regression for fatal/non-fatal cardiovascular disease were: diabetes duration 2.76 (2.21-3.44); onset age 1.47 (1.21-1.78); log ratio total cholesterol:HDL cholesterol 1.26 (1.09-1.45); log HbA(1c) 1.19 (1.03-1.38); log systolic blood pressure 1.17 (1.01-1.34) (1 SD increase in continuous variables); smoker 1.76 (1.27-2.46); macroalbuminuria (> 200 MUg/min) 1.52 (1.10-2.10); previous cardiovascular disease 3.51 (2.54-4.84). All eight variables were used to elaborate a risk equation for 5-year cardiovascular disease risk. Regarding calibration in the derivation data set, ratio predicted 5-year risk (mean 5.4 +/- 7.9%) to observed event rate was 1.0. Discrimination was sufficient, with C statistic 0.83, sensitivity and specificity 72 and 77%, respectively, for the top quartile of predicted risk. Similarly, calibration and discrimination were adequate in the validation data set: ratio of predicted 4-year risk/observed rate 0.94, C-statistic 0.80, sensitivity and specificity 62 and 77%, respectively, for the top quartile. CONCLUSIONS: This 5-year cardiovascular disease risk model from a large observational study of patients with Type 1 diabetes in routine care showed adequate calibration and discrimination and can be useful for clinical practice. It should also be tested in patients with Type 1 diabetes from other countries. PMID- 21627688 TI - Impact of hip circumference and height on incident diabetes: results from 6-year follow-up in the Tehran Lipid and Glucose Study. AB - AIM: To investigate the effects of height and hip circumference, individually and in combination with waist circumference, on incident diabetes. METHODS: A total of 1589 men and 2132 women, free of diabetes at baseline aged >= 20 years participated in a 6-year follow-up examination. The standard 2-h post-challenge plasma glucose test was performed at baseline and during follow-up. The Cox model was used to regress the hazard of diabetes on height and hip circumference individually and in combination with waist circumference. RESULTS: Median follow up was 6 years, with a total of 9433 and 12,607 person-years follow-up among men and women, respectively. The incidence rate of diabetes (95% CIs) was 9.0 (7.6 11.5) and 11.0 (9.6--13.4) per 1000 persons per year in men and women, respectively. Among men, after multiple adjustments, the anthropometric measures were no longer associated with incident diabetes. Among women, the age-adjusted hazard ratio (95% CIs) of diabetes was 0.80 (0.61-1.04) for height, 1.39 (1.21 1.60) for hip circumference and 1.86 (1.59-2.17) for waist circumference. After further adjustment for waist circumference, the hazard ratios were 0.69 (0.53 0.90) for height and 0.76 (0.61-0.96) for hip circumference. These inverse associations resisted multiple adjustments. The hazard ratio for waist circumference increased by 7 and 54% when height and hip circumference were included in the models. CONCLUSION: We observed that height and hip circumference were inversely associated with incident diabetes among women and that these associations could be explored by considering the effect of waist circumference. The risk of diabetes attributable to waist circumference might be underestimated were the effects of height and hip circumference not considered. PMID- 21627689 TI - Recurrent wheeze in children with Down syndrome: is it asthma? AB - AIM: To compare the prevalence of current wheeze in children with Down syndrome (DS), their siblings, and nonrelated population controls. METHODS: This was a case-control study in which the International Study of Asthma and Allergy in Childhood questionnaire for respiratory symptoms was completed by parents for 130 children with DS, 167 of their siblings, and for 119 age- and sex-matched control subjects from the general population. RESULTS: Both wheeze ever and wheeze during the last 12 months was more commonly reported in DS than in their siblings or controls. The relative risk (RR) of current wheeze in DS was 2.8 (95% CI, 1.42 5.51) compared with siblings, and 2.75 (95% CI, 1.28-5.88) compared with controls. A doctor's diagnosis of asthma was found in 3.1% in children with DS, in 4.2% in siblings and in 6.7% in controls. During 4-years follow-up, the diagnosis of asthma could not be confirmed in the 24 DS children with current wheeze, and atopy was found in none of them. CONCLUSION: Wheeze is common in children with DS. This is likely to be related to the factors specific for DS and probably unrelated to asthma. PMID- 21627690 TI - Predictors of death in under-five children with diarrhoea admitted to a critical care ward in an urban hospital in Bangladesh. AB - AIM: To evaluate the clinical and laboratory predictors of death in hospitalized under-five children with diarrhoea. METHODS: This is a prospective cohort study carried out in the Special Care Ward (SCW) of the Dhaka Hospital of the International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR,B), Dhaka, Bangladesh. All admitted diarrhoeal children of both sexes, aged 0-59 months, from September 2007 through December 2007 were enrolled. We compared and analysed factors among diarrhoeal children who died (n = 29) with those who survived (n = 229). RESULTS: In logistic regression analysis, after adjusting for potential confounders (infusion of intravenous fluid and immature PMN), absent peripheral pulse even after complete rehydration (OR 10.9, 95% CI 2.1-56.8; p < 0.01), severe malnutrition (OR 7.9, 95% CI 1.8-34.8; p < 0.01), hypoxaemia (OR 8.5, 95% CI 1.0-75.0; p = 0.05), radiological lobar pneumonia (OR 17.8, 95% CI 3.7-84.5; p < 0.01) and hypernatraemia (OR 15.8, 95% CI 3.0-81.8; p < 0.01) were independently associated with deaths among diarrhoeal children admitted to SCW. CONCLUSIONS: Thus, the absence of peripheral pulses even after full rehydration, severe malnutrition, hypoxaemia, lobar pneumonia and hypernatraemia are independent predictors of death among the under-five children with diarrhoea admitted to critical care ward of a resource-limited setting in Bangladesh. PMID- 21627691 TI - Soft drink and sweet food consumption and suicidal behaviours among Chinese adolescents. AB - AIM: Greater soft drink consumption was associated with increased risk of mental health problems in Western countries. The objective of the study was to examine the association between soft drink and sweet food consumption and suicidal behaviours among adolescents in China. METHODS: In 2005, a population-based cross sectional study was conducted in 100 schools in Jiangsu Province, China. Participants were aged 12-19 years old (n = 23,976). RESULTS: Among the participants, 20.5% reported daily soft drink consumption; 18.6% reported having suicidal ideation. Soft drink consumption was significantly and positively associated with risk of suicidal plan or suicide attempt. Prevalence of suicidal plan was 12.8% among those who consumed soft drink at least three times per day and 6.2% among those who did not consume any soft drinks. In multivariate analyses, compared with soft drink consumption less than once per day, consumption at least three times per day was associated with 80% increased risk for suicidal plan and more than 3.5-fold increased risk for suicide attempt. Of note, nonconsumption of soft drinks was also associated with about 32% elevated risk for suicidal plan and suicidal attempt. High frequency intake of sweet food was associated with increased risk of suicidal behaviours. CONCLUSIONS: There is a positive association between consumption of soft drinks and sweet food and risks for suicidal behaviours among adolescents in China. Prospective studies are warranted to confirm these findings. PMID- 21627692 TI - Waist circumference and waist-to-height ratio in Norwegian children 4-18 years of age: reference values and cut-off levels. AB - AIM: To establish reference values for waist circumference and waist-to-height ratio of Norwegian children. MATERIAL: Data were collected in 2003-2006 as part of a cross-sectional study, including 5725 children 4-18 years of age. Reference curves were fitted with the LMS method; appropriate cut-offs were selected using receiver operating characteristic analysis. RESULTS: Reference values for waist circumference and waist-to-height ratio are presented. Mean waist circumference increased with age for both genders. Boys had a higher waist circumference at almost all ages. Mean waist-to-height ratio decreased until early adolescence and thereafter increased slightly towards adult age. There was a strong positive correlation between waist circumference and BMI (r = 0.907, p < 0.01) and a moderate positive correlation between waist-to-height ratio and BMI (r = 0.397 p < 0.01). A waist circumference cut-off value of 1.0 SDS (85th percentile) gave a sensitivity of 79% and a specificity of 94% to detect overweight. A cut-off value of 1.6 SDS (95th percentile) gave a sensitivity of 94% and a specificity of 96% to detect obesity. CONCLUSION: This study presents the first reference values of waist circumference and waist-to-height ratio for Norwegian children 4-18 years, which also represent the first reference in Scandinavian schoolchildren. The 85th and 95th percentiles of waist circumference are proposed as appropriate cut-offs for central overweight and obesity. PMID- 21627693 TI - Fate of a modified fenestration of atrial septal occluder device after transcatheter closure of atrial septal defects in elderly patients. AB - BACKGROUND: Data on closure of atrial septal defects (ASD) in elderly patients with a fenestrated Amplatzer septal occluder (ASO) device is limited. METHODS: A hemodynamically significant ASD was closed with a fenestrated ASO in 3 patients with ages >62 years. Prior to implant a 4-mm fenestration was created by balloon dilatation without additional suture fixation just adjacent to the stent part of the device. Indications for fenestration were restrictive left ventricular physiology and/or pulmonary hypertension. Heparin had been administered during and for 48 hours after the procedure. Two patients were maintained on phenprocoumon because of chronic atrial fibrillation, the remaining patient on aspirin and clopidogrel for 3 months after implant. Transesophageal echocardiography (TEE) and hemodynamic evaluation were performed 4-18 months after ASD closure. RESULTS: A trace or small fenestration through the ASO with left-to-right shunt was detected by TEE in all 3 patients without any hemodynamic significance. No thrombus formation was observed. Pulmonary hypertension improved in the affected patient. Pulmonary arterial wedge pressure and cardiac index improved in the second patient with improvement in heart failure symptoms and of quality of life in both. The third patient, after initial improvement for 6 months, developed significant comorbidities and clinical deterioration at 18 months follow-up. CONCLUSION: The modified fenestration of the ASO decreased significantly in size at follow-up. Applying this technique to selected patients judged to be at risk for ASD closure avoids acute decompensation and allows gradual diminuition of right ventricular volume overload during mid-term follow up. PMID- 21627694 TI - Immunohistochemical assessment of CD1a-positive Langerhans cells and their relationship with E-cadherin in minor salivary gland tumors. AB - OBJECTIVE: The aim of this study was to investigate the presence of CD1a-positive Langerhans cells and their relationship with E-cadherin in minor salivary gland tumors. METHODS: Twenty-seven minor salivary gland tumors were investigated using immunohistochemistry for CD1a and E-cadherin. RESULTS: A significant difference regarding the mean density of CD1a-positive Langerhans cells was observed between pleomorphic adenomas and malignant tumors studied (P = 0.001). No CD1a-positive cells were detected in most cases (n = 5) of cystic adenoid carcinomas. CD1a positive cells were detected in one mucoepidermoid carcinoma case, and six low grade polymorphous adenocarcinomas cases. Comparison of the mean density of CD1a positive cells between the three malignant tumors showed no significant difference (P = 0.127). No significant difference was observed in the presence of E-cadherin between tumors (P = 0.73), but it was detected in 24 cases. CONCLUSIONS: The lack of CD1a-positive in malignant salivary gland tumors facilitates the neoplastic development and suggests that these cells might be useful as auxiliary diagnostic and prognostic tool in minor salivary gland tumors. Furthermore, it is suggested that E-cadherin mediates cell adhesion in these tumors although we did not demonstrate significance. PMID- 21627695 TI - An outbreak of Fusarium solani endophthalmitis after cataract surgery in an eye training and research hospital in Istanbul. AB - To report an outbreak of Fusarium solani endophthalmitis after uneventful cataract surgeries performed on the same day in the same operating room. Nine patients underwent phacoemulsification at 4th Clinic of Beyoglu Eye Training and Research Hospital in Istanbul. Cefuroxime axetyl was injected intracamerally from the same vial to all patients at the end of surgery. All patients developed acute postoperative endophthalmitis. Presentation, cultural studies, treatment, clinical responses and risk factors were evaluated. Cultural and DNA sequence findings revealed F. solani. Antifungal therapy was begun and pars plana vitrectomy, intraocular lens and capsule extraction were performed. Corneal involvement was correlated with old age and systemic disease. Fusarium solani should be considered in acute postoperative endophthalmitis. This infection can be controlled with early and aggressive combined antifungal and surgical treatment. The patients with corneal involvement had poor prognosis. It is important to use solutions prepared separately for each patient. PMID- 21627696 TI - Comparison of exendin-4 on beta-cell replication in mouse and human islet grafts. AB - Exendin-4 can stimulate beta-cell replication in mice. Whether it can stimulate beta-cell replication in human islet grafts remains unknown. Therefore, we compared the effects of exendin-4 on beta-cell replication in mouse and human islet grafts. Islets, isolated from mouse and human donors at different ages, were transplanted into diabetic mice and/or diabetic nude mice that were given bromodeoxyuridine (BrdU) with or without exendin-4. At 4 weeks post transplantation, islet grafts were removed for insulin and BrdU staining and quantification of insulin(+)/BrdU(+) cells. Although diabetes was reversed in all mice transplanting syngeneic mouse islets from young or old donors, normoglycemia was achieved significantly faster in exendin-4 treated mice. Mouse islet grafts in exendin-4 treated mice had significantly more insulin(+)/BrdU(+) beta cells than in untreated mice (P < 0.01). Human islet grafts from <=22-year-old donors had more insulin(+)/BrdU(+) beta cells in exendin-4 treated mice than that in untreated mice (P < 0.01). However, human islet grafts from >=35-year-old donors contained few insulin(+)/BrdU(+) beta cells in exendin-4 treated or untreated mice. Our data demonstrated that the capacity for beta-cell replication in mouse and human islet grafts is different with and without exendin-4 treatment and indicated that GLP-1 agonists can stimulate beta-cell replication in human islets from young donors. PMID- 21627697 TI - Tackling the controversial topics in neuroendocrinology. PMID- 21627698 TI - Exploring risk factors for depression among older men residing in Macau. AB - AIM: To determine the prevalence of depression level of older men in Macau and identify factors that predict depression. BACKGROUND: Studies have revealed the contribution of psychosocial factors such as stress, dissatisfaction at work and stressful life events to older men and the mental health burdens pose a grave threat to their quality of life. DESIGN: A descriptive survey. METHOD: The study was conducted from July-August 2004 in six parishes in Macau; 839 older men aged 60+ completed a structured questionnaire. OUTCOME MEASURES: Predisposing characteristics, social and daily activities, health history and need/behaviours and depression level were collected. Multiple logistic regression models were used to identify factors to predict older men who will have depression. RESULTS: The prevalence rate of depression of older men was 8.6%. It showed that a history of stroke (p=0.039), insomnia (p<0.001), palpitations (p=0.014), poor social network (p=0.005) and self-perceived health status (p=0.001) and perceived low ability to meet living costs (p<0.001) were significant risk factors for depression. CONCLUSION: We should focus on older men by reducing their burden to meet living costs, improving their sleep quality and helping them to expand their social network. These should help in both prevention and recognition of the onset of depression. Those with the low social network scores could be targeted for more intensive support from the beginning. RELEVANCE TO CLINICAL PRACTICE: It is important to screen regularly for depression status among older men in the community. Attention to poor perceived health and not enough on the ability to meet living costs were needed and important to follow up. PMID- 21627699 TI - Electronic prescribing reduces prescribing error in public hospitals. AB - AIMS AND OBJECTIVES: To examine the incidence of prescribing errors in a main public hospital in Pakistan and to assess the impact of introducing electronic prescribing system on the reduction of their incidence. BACKGROUND: Medication errors are persistent in today's healthcare system. The impact of electronic prescribing on reducing errors has not been tested in developing world. DESIGN: Prospective review of medication and discharge medication charts before and after the introduction of an electronic inpatient record and prescribing system. METHODS: Inpatient records (n = 3300) and 1100 discharge medication sheets were reviewed for prescribing errors before and after the installation of electronic prescribing system in 11 wards. RESULTS: Medications (13,328 and 14,064) were prescribed for inpatients, among which 3008 and 1147 prescribing errors were identified, giving an overall error rate of 22.6% and 8.2% throughout paper-based and electronic prescribing, respectively. Medications (2480 and 2790) were prescribed for discharge patients, among which 418 and 123 errors were detected, giving an overall error rate of 16.9% and 4.4% during paper-based and electronic prescribing, respectively. CONCLUSION: Electronic prescribing has a significant effect on the reduction of prescribing errors. RELEVANCE TO CLINICAL PRACTICE: Prescribing errors are commonplace in Pakistan public hospitals. The study evaluated the impact of introducing electronic inpatient records and electronic prescribing in the reduction of prescribing errors in a public hospital in Pakistan. PMID- 21627700 TI - The emotional sequelae of whistleblowing: findings from a qualitative study. AB - AIMS AND OBJECTIVES: To highlight and illuminate the emotional sequelae of whistleblowing from whistleblowers and subjects of whistleblowing complaints. BACKGROUND: Whistleblowing has the potential to have a negative impact on individuals' physical and emotional well-being. However, few empirical studies have been conducted using qualitative methods to provide an in-depth exploration of the emotional consequences for those involved in whistleblowing incidents. DESIGN: Qualitative narrative inquiry design. METHOD: Purposive sampling was used to recruit participants who had been involved in whistleblowing incidents. During interviews participants' accounts were digitally recorded and then transcribed verbatim. Data were then analysed by two researchers until consensus was reached. RESULTS: Findings revealed that participants' emotional health was considerably compromised as a result of the whistleblowing incident. Analysis of the data revealed the following dominant themes: 'I felt sad and depressed': overwhelming and persistent distress; 'I was having panic attacks and hyperventilating': acute anxiety; and, 'I had all this playing on my mind': nightmares, flashbacks and intrusive thoughts. CONCLUSIONS: While it has been previously acknowledged that whistleblowing has the potential to have a negative impact on all aspects of an individual's life, this study notably highlights the intensity of emotional symptoms suffered by participants as well as the extended duration of time these symptoms were apparent. RELEVANCE TO CLINICAL PRACTICE: As professionals, nurses, as well as organisations, have a responsibility to identify those who may be suffering the emotional trauma of whistleblowing and ensure they have access to appropriate resources. PMID- 21627701 TI - The dilemma of diabetic patients living with hypoglycaemia. AB - AIMS: To examine the impact of the threat of hypoglycaemic episodes on people with diabetes in Taiwan. BACKGROUND: Intensive diabetes treatment in people with diabetes helps them to achieve better glycaemic control. However, it also causes more frequent hypoglycaemic episodes and has an impact on their overall quality of life. Hypoglycaemia is accompanied by various distressing symptoms which may cause excessive fear, affecting decision making in hypoglycaemic management. DESIGN: Purposive sampling and in-depth, face-to-face interviews were used to collect data. METHODS: Semi-structured interviews were conducted from July 2008 January 2009 with 17 individuals treated with insulin who had previous hypoglycaemic episodes. Data were analysed using qualitative content analysis. RESULTS: Four themes were generated from the analysis, 'inability to control fluctuations in health', 'challenges to interpersonal relationships', 'facing the disease alone' and 'finding a balance between competing symptoms'. CONCLUSIONS: Hypoglycaemia is a major health issue for many people with diabetes. Understanding individuals' experiences with hypoglycaemic episodes should help practitioners become more fully involved in promoting self-management. We identified key areas that health care providers should address, including concerns about patient education and professional support for people with diabetes experiencing hypoglycaemia, to enhance problem solving skills for them and their families. RELEVANCE TO CLINICAL PRACTICE: We recommend that health care providers make proper use of support groups for family caregivers or other important individuals in the lives of people with diabetes to provide education, clarification, support and guidance. In addition, health care providers also need to provide clients with hypoglycaemia-related emotional support, while enhancing diabetes self-management and problem-solving skills. PMID- 21627702 TI - The Bantu expansion revisited: a new analysis of Y chromosome variation in Central Western Africa. AB - The current distribution of Bantu languages is commonly considered to be a consequence of a relatively recent population expansion (3-5kya) in Central Western Africa. While there is a substantial consensus regarding the centre of origin of Bantu languages (the Benue River Valley, between South East Nigeria and Western Cameroon), the identification of the area from where the population expansion actually started, the relation between the processes leading to the spread of languages and peoples and the relevance of local migratory events remain controversial. In order to shed new light on these aspects, we studied Y chromosome variation in a broad dataset of populations encompassing Nigeria, Cameroon, Gabon and Congo. Our results evidence an evolutionary scenario which is more complex than had been previously thought, pointing to a marked differentiation of Cameroonian populations from the rest of the dataset. In fact, in contrast with the current view of Bantu speakers as a homogeneous group of populations, we observed an unexpectedly high level of interpopulation genetic heterogeneity and highlighted previously undetected diversity for lineages associated with the diffusion of Bantu languages (E1b1a (M2) sub-branches). We also detected substantial differences in local demographic histories, which concord with the hypotheses regarding an early diffusion of Bantu languages into the forest area and a subsequent demographic expansion and migration towards eastern and western Africa. PMID- 21627703 TI - On the specificity of avian blood parasites: revealing specific and generalist relationships between haemosporidians and biting midges. AB - The study of host-parasite relationships involving vector-borne parasites requires understanding interactions between parasites and vectors. The capacity of haemosporidians to infect insects has clear evolutionary consequences for the transmission of diseases. Here, we investigated (i) the associations between blood parasites, biting midges and birds and (ii) the potential specificity between biting midge and haemosporidian haplotypes. A total of 629 parous biting midges Culicoides and 224 wild birds (belonging to seven species) from a locality of central Spain were individually examined for the presence of Haemoproteus and Plasmodium parasites by sequencing a fragment of cytochrome B. Biting midges were identified morphologically and characterized on the basis of a fragment of the cytochrome c oxidase (COI) gene. Overall, 12 Haemoproteus and three Plasmodium haplotypes were isolated and sequenced. Among them, 10 haplotypes were exclusively isolated from biting midges, three haplotypes only from birds and two haplotypes from both biting midges and birds. Biting midge haplotypes showed both specific and generalist relationships with Haemoproteus haplotypes but only generalist relationships with Plasmodium haplotypes. Several C. festivipennis and C. kibunesis haplotypes established significant coevolutionary links with Haemoproteus haplotypes. These results shed light on the specificity of interactions between vectors and blood parasites. PMID- 21627704 TI - The genetic structure of the plant pathogenic fungus Melampsora larici-populina on its wild host is extensively impacted by host domestication. AB - Wild and cultivated plants represent very different habitats for pathogens, especially when cultivated plants bear qualitative resistance genes. Here, we investigated to what extent the population genetic structure of a plant pathogenic fungus collected on its wild host can be impacted by the deployment of resistant cultivars. We studied one of the main poplar diseases, poplar rust, caused by the fungus Melampsora larici-populina. A thousand and fifty individuals sampled from several locations in France were phenotyped for their virulence profile (ability to infect or not the most deployed resistant cultivar 'Beaupre'), and a subset of these was genotyped using 25 microsatellite markers. Bayesian assignment tests on genetic data clustered the 476 genotyped individuals into three genetic groups. Group 1 gathered most virulent individuals and displayed evidence for selection and drastic demographic changes resulting from breakdown of the poplar cultivar 'Beaupre'. Group 2 comprised individuals corresponding to ancestral populations of M. larici-populina naturally occurring in the native range. Group 3 displayed the hallmarks of strict asexual reproduction, which has never previously been demonstrated in this species. We discuss how poplar cultivation has influenced the spatial and genetic structure of this plant pathogenic fungus, and has led to the spread of virulence alleles (gene swamping) in M. larici-populina populations evolving on the wild host. PMID- 21627705 TI - Real world evaluation of dual-zone ICD and CRT-D programming compared to single zone programming: the ALTITUDE REDUCES study. AB - INTRODUCTION: We evaluated the frequency of appropriate and inappropriate shocks and survival in patients using dual-zone programming versus single-zone programming. METHODS AND RESULTS: For the ALTITUDE REDUCES study, patients were followed for 1.6 +/- 1.1 years. The 12-month incidence of any shock was lower for dual-versus single-zone programmed detection at rates <=170 bpm and between 170 200 bpm (P < 0.001). Appropriate shock rates at 1 year were also lower with dual zone programming in these rate intervals (single zone 9.1%, 5.4%, P < 0.001, dual zone 6.7%, 4.7%, P < 0.02). There were no detectable differences between single- and dual-zone shock incidence at detection rates >= 200 bpm (P = 0.14). Inappropriate shock incidence was less with dual- versus single-zone detection at all detect rates <200 bpm, but not at rates >=200 bpm (P < 0.001, P = 0.37). The lowest risk of appropriate and inappropriate shock was associated with dual-zone programming and detection rates >=200 bpm (2.1%). Dual-zone detection was associated with more nonsustained and diverted therapy episodes but these patients did not have an increased risk of death compared to patients with single zone programming. Patients programmed to low detection rate, single-zone detection and shock-only therapy also had the highest preshock mortality risk (P = 0.05). CONCLUSIONS: Shock incidence is lowest with either single- or dual-zone detection >=200 bpm. For detection rates <200 bpm, dual-zone programming is associated with a reduction in the incidence of total shocks, appropriate shocks, and inappropriate shocks. PMID- 21627706 TI - Physical properties of an acrylic resin after incorporation of an antimicrobial monomer. AB - PURPOSE: This study evaluated the effect of the incorporation of the antimicrobial monomer methacryloyloxyundecylpyridinium bromide (MUPB) on the hardness, roughness, flexural strength, and color stability of a denture base material. MATERIALS AND METHODS: Ninety-six disk-shaped (14-mm diameter * 4-mm thick) and 30 rectangular (65 * 10 * 3.3 mm(3) ) heat-polymerized acrylic resin specimens were divided into three groups according to the concentration of MUPB (w/w): (A) 0%, (B) 0.3%, (C) 0.6%. Hardness was assessed by a hardness tester equipped with a Vickers diamond penetrator. Flexural strength and surface roughness were tested on a universal testing machine and a surface roughness tester, respectively. Color alterations (DeltaE) were measured by a portable spectrophotometer after 12 and 36 days of immersion in water, coffee, or wine. Variables were analyzed by ANOVA/Tukey HSD test (alpha= 0.05). RESULTS: The following mean results (+/-SD) were obtained for hardness (A: 15.6 +/- 0.6, B: 14.6 +/- 1.7, C: 14.8 +/- 0.8 VHN; ANOVA: p= 0.061), flexural strength (A: 111 +/ 17, B: 105 +/- 12, C: 88 +/- 12 MPa; ANOVA: p= 0.008), and roughness (A: 0.20 +/ 0.11, B: 0.20 +/- 0.11, C: 0.24 +/- 0.08 MUm; ANOVA: p= 0.829). Color changes of immersed specimens were significantly influenced by solutions and time (A: 9.1 +/ 3.1, B: 14.8 +/- 7.5, C: 13.3 +/- 6.1 DeltaE; ANOVA: p < 0.05). CONCLUSIONS: The incorporation of MUPB affects the mechanical properties of a denture base acrylic resin; however, the only significant change was observed for flexural strength and may not be critical. Color changes were slightly higher when resin containing MUPB was immersed in wine for a prolonged time; however, the difference has debatable clinical relevance. PMID- 21627707 TI - Effect of embedded metal reinforcements and their location on the fracture resistance of acrylic resin complete dentures. AB - PURPOSE: This study evaluated the effect of metal reinforcement and its location on the flexural load at the proportional limit (FL-PL) and the flexural deflection of maxillary acrylic resin complete dentures. MATERIALS AND METHODS: Maxillary acrylic resin complete dentures reinforced with Remanium and without reinforcement were tested. The reinforcing material was embedded in the denture base resin in the doughy state and placed (1) under the ridge lap region; (2) in the anterior region; (3) in the middle region; and (4) in the anterior and posterior regions. The FL-PL (N) and the flexural deflection (mm) at 100 N of the reinforced maxillary denture specimens were tested using a load testing machine at a 5.0 mm/min crosshead speed. The data were analyzed statistically using one way ANOVA; Tukey's post hoc comparisons test was applied when appropriate (95% confidence level). RESULTS: The FL-PL of the dentures without reinforcement (909 +/- 195 N) and the dentures reinforced at the ridge lap (1094 +/- 176 N) and in the middle (977 +/- 215 N) regions were not significantly different (p > 0.05). The dentures reinforced in the anterior (1348 +/- 205 N) and the anterior and posterior (1190 +/- 191 N) regions had a higher FL-PL than the dentures without reinforcement (p < 0.05) and were not significantly different from each other (p > 0.05). The efficiency (times) of the reinforcing material on the dentures without reinforcement was 1.08 to 1.48. The flexural deflection of the dentures without reinforcement (0.133 +/- 0.014 mm), the dentures reinforced at the ridge lap (0.125 +/- 0.014 mm), in the anterior (0.122 +/- 0.009 mm), and in the middle (0.132 +/- 0.015 mm) regions were not significantly different (p > 0.05), and the dentures reinforced in the anterior and posterior (0.117 +/- 0.011 mm) regions had significantly lower deflection than the dentures without reinforcement (p < 0.05). CONCLUSION: The location of the metal reinforcement affected the fracture resistance of the maxillary acrylic resin complete dentures. PMID- 21627708 TI - Influence of zirconia base and shade difference on polymerization efficiency of dual-cure resin cement. AB - PURPOSE: The aim of this study was to investigate the polymerization efficiency of dual-cured resin cement beneath different shades of zirconia-based feldsphathic ceramic restorations. MATERIALS AND METHODS: Five translucent zirconia (Zirkonzahn) discs (4.0-mm diameter, 1.2-mm height) were prepared. Feldsphathic ceramic (1.2 mm) (Noritake Cerabien Zr) in 5 shades (1M2, 2M2, 3M2, 4M2, 5M2) was applied on the zirconia discs. Twelve dual-cure resin cement specimens were prepared for each shade, using Panavia F 2.0 (Kuraray) in Teflon molds (4.0-mm diameter, 6.0-mm height), following the manufacturer's instructions. Light activation was performed through the zirconia-based ceramic discs for 20 seconds, using a quartz tungsten halogen curing device (Hilux 200) with irradiance of 600 mW/cm(2) . Immediately following light curing, specimens were stored for 24 hours in dry, light-proof containers. Vickers hardness measurements were conducted using a microhardness tester with a 50-g load applied for 15 seconds. The indentations were made in the cross sectional area at four depths, and the mean values were recorded as Vickers hardness number (VHN). Results were statistically analyzed with one-way ANOVA and Tukey HSD test (p < 0.05). RESULTS: A statistically significant decrease in VHN of the resin cement was noted with increasing depth and darkness of the shade (p < 0.05). CONCLUSION: Curing efficiency of dual-cure resin cement is mainly influenced by the lightness of the shades selected. PMID- 21627709 TI - Canine tip marker: a simplified tool for measuring intercanine distance. AB - PURPOSE: Appropriate selection of anterior teeth is considered to be of paramount importance in the success of denture prostheses. This study was undertaken to evaluate the correlation between the intercanthal width and interalar width with intercanine distance, in North Indian male and female patients for predicting the mesiodistal width of the maxillary anterior teeth during tooth selection. MATERIALS AND METHODS: The study was conducted with 100 North Indian patients (50 men, 50 women) ranging in age from 17 to 21 years. A digital caliper with an accuracy of 0.01 mm was used to measure the intercanthal and interalar width. A T shaped flat metal plate (canine tip marker) was used to mark the intercanine distance, which was then measured with the digital caliper. These measurements were interpreted and subjected to statistical analysis. Student's t-test was applied to test the correlation between intercanthal width and interalar width with intercanine distance. RESULTS: Calculated t-values between intercanine distances with interalar width in both male and female groups were 3.14 and 3.56, respectively, greater than the standard value taken at a 5% level of significance with 48 degree of freedom, showing a higher correlation of interalar width with the intercanine distance. Values obtained between intercanthal width and intercanine distance were lower than the standard value in both groups. CONCLUSIONS: A significant correlation was found between interalar width and intercanine distance in both men and women, suggesting that interalar width can be used as a reliable guide for maxillary anterior teeth selection. PMID- 21627710 TI - Effect of dentin sealers on postoperative sensitivity of complete cast crowns cemented with glass ionomer cement. AB - PURPOSE: The purpose of this study was to clinically evaluate the effects of pretreatments with copal/ether varnish and dentin bonding system on postoperative sensitivity of complete cast crowns cemented with glass ionomer cement. MATERIALS AND METHODS: Three posterior teeth with no pain symptoms were selected from each of 17 patients, totaling 51 teeth, for which a crown was indicated. Rexillium III complete cast crowns were prepared using conventional laboratory techniques. For each patient, the first tooth, which served as the control, received only glass ionomer cement (Ketac-Cem). Copal/ether varnish (Bosworth Copaliner) was applied to the second tooth preparation prior to cementation. Dentin bonding agent (OptiBond Solo Plus) was used on the third tooth before cementation. Sensitivity to different stimuli (cold, heat) was assessed at 7 days, 1 month, and 6 months following restorative procedures by questionnaire. RESULTS: There were no statistically significant differences between the three groups regarding applied stimulus and day of the study (p > 0.05). No statistically significant differences were found between the postoperative sensitivity responses from 7 days to 1 month, and from 1 month to 6 months (p > 0.05). CONCLUSIONS: Postoperative sensitivity resulting from glass ionomer cement with complete cast crowns cannot be completely eliminated with the prior use of a cavity varnish or bonding agent. PMID- 21627711 TI - Topical prolyl hydroxylase domain-2 silencing improves diabetic murine wound closure. AB - Prolyl hydroxylase domain 2 (PHD2) has been implicated in several pathways of cell signaling, most notably in its regulation of hypoxia-inducible factor (HIF) 1alpha stability. In normoxia, PHD2 hydroxylates proline residues on HIF-1alpha, rendering it inactive. However, in hypoxia, PHD2 is inactive, HIF-1alpha is stabilized and downstream effectors such as vascular endothelial growth factor and fibroblast growth factor-2 are produced to promote angiogenesis. In the present study we utilize RNA interference to PHD2 to promote therapeutic angiogenesis in a diabetic wound model, presumably by the stabilization of HIF 1alpha. Stented wounds were created on the dorsum of diabetic Lepr db/db mice. Mice were treated with PHD2 small interfering RNA (siRNA) or nonsense siRNA. Wounds were measured photometrically on days 0-28. Wounds were harvested for histology, protein, and RNA analysis. Diabetic wounds treated with siRNA closed within 21+/-1.2 days; sham-treated closed in 28+/-1.5 days. By day 7, Western blot revealed near complete suppression of PHD protein and corresponding increased HIF-1alpha. Angiogenic mediators vascular endothelial growth factor and fibroblast growth factor-2 were elevated, corresponding to increased CD31 staining in the treated groups. siRNA-mediated silencing of PHD2 increases HIF 1alpha and several mediators of angiogenesis. This corresponded to improved time to closure in diabetic wounds compared with sham-treated wounds. These findings suggest that impaired wound healing in diabetes can be ameliorated with therapeutic angiogenesis. PMID- 21627712 TI - Angiogenesis in mesenteric microvascular networks from spontaneously hypertensive versus normotensive rats. AB - OBJECTIVE: Elevated blood pressure during hypertension has been associated with microvascular rarefaction, defined as a loss of microvessels. However, whether rarefaction is a result of impaired angiogenesis remains unclear. The objective of this study was to compare angiogenesis across the time course of mesenteric microvascular network remodeling in adult spontaneously hypertensive versus normotensive rats. METHODS: Angiogenic responses in 15- to 16-week-old SHR and Wistar rats at 0, 3, 5, 10 or 25 days post 20-minute exteriorization of the mesentery were quantified. RESULTS: Consistent with the phenomenon of rarefaction, vascularized area in unstimulated SHR was decreased compared to Wistar. By 25 days, SHR vascular area had increased to the Wistar level and vascular length density and capillary sprouting were comparable. At 3 and 5 days, SHR and Wistar tissues displayed an increase in the capillary sprouting and vascular density relative to their unstimulated controls. At 10 days, capillary sprouting in the SHR remained elevated. The percent change in vascular density was elevated in the SHR compared to the Wistar group at 3 and 5 days and by 25 days the rate of change was more negative. CONCLUSIONS: Our results suggest that SHR networks undergo an increased rate of growth followed by an increased rate of pruning. PMID- 21627713 TI - Risk, Error, Outcome, and Prevention in Pediatric Anesthesia: so many issues, lots of good solutions, but where do we find the resources? PMID- 21627714 TI - Ultrasound-guided subclavian vein cannulation in infants: supraclavicular approach. AB - BACKGROUND: Ultrasound (US) guidance techniques are reported to be safe for internal jugular vein catheterization, although anatomic conditions are not favorable for this approach in infants. The subclavian vein (SCV) seems to be a better site for long-term central venous catheterization in children, with a supraclavicular approach to avoid compression of the central venous catheter between the clavicle and the first rib ('pinch-off' syndrome). We describe a new US-guided approach for supraclavicular SCV cannulation in infants. METHODS: The principle of this technique is to place the US probe at the supraclavicular level to obtain a longitudinal view of the SCV, and to gain access to the vein with a total ultrasonic control (in-plane puncture) via a supraclavicular approach known since 1965, but rarely used in blind puncture. The results of 37 US-guided SCV cannulations in infants weighing <10 kg are reported. RESULTS: Forty-two infants were enrolled in this observational study, and five infants with bad visualization of SCV were excluded. The procedure duration was <5 min in all cases except one. The success rate at the first attempt was 81% and 100% after two attempts. No major complications were reported. CONCLUSIONS: This US-guided supraclavicular approach for SCV puncture is a new possibility for central venous catheterization in small infants, offering all the advantages of SCV cannulation without the risk of 'pinch-off' syndrome. This technique seems valuable for children and infants and quite easy to apply for physicians trained to US guidance punctures. PMID- 21627715 TI - Pandemic response lessons from influenza H1N1 2009 in Asia. AB - During April 2009, a novel H1N1 influenza A virus strain was identified in Mexico and the USA. Within weeks the virus had spread globally and the first pandemic of the 21st Century had been declared. It is unlikely to be the last and it is crucial that real lessons are learned from the experience. Asia is considered a hot spot for the emergence of new pathogens including past influenza pandemics. On this occasion while preparing for an avian, highly virulent influenza virus (H5N1 like) originating in Asia in fact the pandemic originated from swine, and was less virulent. This discrepancy between what was planned for and what emerged created its own challenges. The H1N1 pandemic has tested national health-care infrastructures and exposed shortcomings in our preparedness as a region. Key health challenges include communication throughout the region, surge capacity, access to reliable information and access to quality care, health-care worker skills, quality, density and distribution, access to essential medicines and lack of organizational infrastructure for emergency response. Despite years of preparation the public health and clinical research community were not ready to respond and opportunities for an immediate research response were missed. Despite warm words and pledges efforts to engage the international community to ensure equitable sharing of limited resources such as antivirals and vaccines fell short and stockpiles in the main remained in the rich world. This manuscript with authors from across the region describes some of the major challenges faced by Asia in response to the pandemic and draws lessons for the future. PMID- 21627716 TI - Changing epidemiology of respiratory pathogens and the role of improved diagnostics. PMID- 21627718 TI - A complex Aboriginal health project and the challenges for evaluation. PMID- 21627719 TI - Physical and mental health status of soldiers responding to the 2008 Wenchuan earthquake. AB - OBJECTIVE: While internationally major disasters occur frequently, for any one country they are unusual events. In this project we aimed to identify public health issues arising from the physical and mental health symptoms suffered by the soldier volunteers deployed in an emergency relief task during the Wenchuan earthquake. METHODS: Health problems identified in other emergency volunteer populations guided the development of a questionnaire. A cohort of 1,187 soldier volunteers completed the questionnaire, which measured physical and mental health symptoms during their rescue mission. The results were compared with a population norm of soldiers, although baseline data of the respondents were unavailable. RESULTS: Half the respondents reported suffering from skin and mucous membrane problems, followed by respiratory symptoms (38%), digestive (29%) and nervous (22%) symptoms. Despite a low response rate (53%) to the mental health component, nearly half (49%) of those who did respond reported mental health problems. The incidence of the above symptoms were significantly higher than the general soldier population. CONCLUSIONS: Health complaints were common in the soldiers, who had not received any formal training in rescue operations. IMPLICATIONS: Non professional rescue workers who are not appropriately prepared for the role may suffer more than their professional counterparts. Attention needs to be paid to the health and safety of non-professional rescue workers, which has been ignored in most disaster management plans. These findings can be used to enhance the understanding of emergency response programs within and outside China, where this particular disaster occurred. PMID- 21627720 TI - Trends in oral, pharyngeal and oesophageal cancer mortality in Australia: the comparative importance of tobacco, alcohol and other risk factors. AB - OBJECTIVE: The relationship of long-term population-level trends in oral, pharyngeal and oesophageal cancer mortality with major risk factors such as tobacco consumption have not been statistically analysed in Australia. We have demonstrated the long-term implications using historical data. METHODS: Estimated age and sex-specific tobacco consumption back-extrapolated to 1887 were used together with alcohol and fruit and vegetable consumption data to examine their association with trends in oral, pharyngeal and oesophageal cancer mortality. Log linear Poisson regression models were applied to specify the relationship with oesophageal and pharyngeal mortality data. RESULTS: Oral cancer mortality for males decreased sharply in the first half of the 20th Century in contrast to steadily rising tobacco consumption. Female oral and pharyngeal cancer remained steady at low levels. Post-World War II male and female oesophageal and male pharyngeal cancer mortality rose, then either fell or stabilised, without a clear relationship with risk factors. CONCLUSIONS: Tobacco and alcohol consumption have influenced post-World War II trends in oral, pharyngeal and oesophageal cancer mortality. However, the challenges in using historical population level data prevent precise interpretation of findings. IMPLICATIONS: There is increased exposure to risk factors for these cancers in many low- and middle-income countries. In particular, smoking cessation programs are needed to prevent increases in mortality from these cancers in such countries. PMID- 21627721 TI - Increasing cancer mortality among opioid-dependent persons in Australia: a new public health challenge for a disadvantaged population. AB - OBJECTIVE: To examine cancer mortality in a population-based cohort of opioid dependent persons. METHODS: New South Wales opioid substitution therapy (OST) program registrants from 1985 to 2005 (n=43,789) were probabilistically linked to the National Death Index. Crude and standardised mortality rates and standardised mortality ratios (SMRs) were calculated. RESULTS: The crude cancer mortality rate increased from 4 to 65 deaths per 100,000 person-years (p trend <0.001). Overall, OST registrants were 1.7 times more likely to die of cancer than the general population (SMR 95% CI 1.4-1.9). Site-specific SMRs were significantly elevated for lung cancer (3.6, 95% CI 2.8-4.6), liver cancer (6.9, 95% CI 4.3-10.5), and anogenital cancers (2.8, 95% CI 1.3-5.3), and significantly reduced for breast cancer (0.4, 95% CI 0.1-0.9). CONCLUSIONS: Cancer is an increasingly important cause of death among OST registrants as they live longer with their dependency. The site-specific excess deaths suggest the role of tobacco, alcohol, and infection with hepatitis C and human papillomavirus. IMPLICATIONS: The OST setting may be a useful setting for the delivery of programs aimed at detection of precursor lesions, reducing exposure to established carcinogens, and treatment for those with HCV infection. Such targeted steps are likely to reduce the future cancer burden in this population. PMID- 21627722 TI - Compliance with professional guidelines with reference to familial cancer services. AB - OBJECTIVE: Professional guidelines define the risk categorisation of patients for a genetic predisposition to cancer based on family history. These guidelines inform the appropriate referral of patients to specialist familial cancer services. Our study aimed to determine the quality of referral letters from general practitioners and specialists to genetic services for breast, ovarian and colorectal cancers, and their compliance with relevant professional guidelines. METHODS: A retrospective review of the referral letters and patient files of 241 consecutive patients referred between June and October 2008. RESULTS: Sufficient information to make a risk assessment was provided in 71% of referrals. Of these, 89% were compliant with guidelines. Genetic counsellors collected further information on 167 of the 241 referred patients and of these 83% were appropriate for referral according to guidelines. CONCLUSIONS AND IMPLICATIONS: Overall, referrals to familial cancer genetic services complied with professional referral guidelines. The majority of referrals were high quality, and with additional information, most patients were shown to be appropriate for review in a familial cancer clinic. Despite this, a better understanding of the reasons for non compliant referrals, and appropriate targeted education and resources is recommended to improve referral quality and compliance. PMID- 21627723 TI - Racism, social resources and mental health for Aboriginal people living in Adelaide. AB - BACKGROUND: This paper examines whether reported experience of racism by Aboriginal people living in Adelaide is negatively associated with mental health, and whether social resources ameliorate the mental health effects of racism. METHODS: Face-to-face structured and semi-structured interviews were conducted with 153 Aboriginal people. Data on self-reported experiences of racism (average regularity of racism across a number of settings, regular racism in at least one setting), social resources (socialising, group membership, social support, talking/expressing self about racism), health behaviours (smoking, alcohol), socio-demographic (age, gender, education, financial situation) and mental health (SF-12 measure) are reported. Separate staged linear regression models assessed the association between the two measures of racism and mental health, after accounting for socio-demographic characteristics and health behaviours. Social resource variables were added to these models to see if they attenuated any relationship between racism and mental health. RESULTS: The two measures of racism were negatively associated with mental health after controlling for socioeconomic factors and health behaviours. These relationships remained after adding social resource measures. Non-smokers had better mental health, and mental health increased with positive assessments of financial situation. CONCLUSION AND IMPLICATIONS: Reducing racism should be a central strategy in improving mental health for Aboriginal people. PMID- 21627724 TI - Successful chronic disease care for Aboriginal Australians requires cultural competence. AB - OBJECTIVE: To review the literature to determine the attributes of culturally appropriate healthcare to inform the design of chronic disease management (CDM) models for Aboriginal patients in urban general practice. METHODS: A comprehensive conceptual framework, drawing on the Access to Care, Pathway to Care, Chronic Care, Level of Connectedness, and Cultural Security, Cultural Competency and Cultural Respect models, was developed to define the search strategy, inclusion criteria and appraisal methods for the literature review. Selected papers were reviewed in detail if they examined a chronic disease intervention for an Aboriginal population and reported on its evaluation, impacts or outcomes. RESULTS: In the 173 papers examined, only 11 programs met the inclusion criteria. All were programs conducted in rural and remote Aboriginal community-controlled health services. Successful chronic disease care and interventions require adequate Aboriginal community engagement, utilising local knowledge, strong leadership, shared responsibilities, sustainable resources and integrated data and systems. These success factors fitted within the conceptual framework developed. CONCLUSIONS: Research and development of culturally appropriate CDM models concurrently in both urban and rural settings will enable more rigorous evaluation, leading to stronger evidence for best practice. A partnership of mainstream and Aboriginal-controlled health services is essential to successfully 'close the gap'. IMPLICATIONS: Findings will inform and guide the development, implementation and evaluation of culturally appropriate CDM in mainstream general practice and primary care. PMID- 21627725 TI - A Kaupapa Maori approach to a community cohort study of heart disease in New Zealand. AB - OBJECTIVE: To report the processes and protocols that were developed in the design and implementation of the Hauora Manawa Project, a cohort study of heart disease in New Zealand and to report the participation at baseline. METHODS: This study utilised application of a Kaupapa Maori Methodology in gaining tribal and health community engagement, design of the project and random selection of participants from territorial electoral rolls, to obtain three cohorts: rural Maori, urban Maori and urban non-Maori. Logistic regression was used to model response rates. RESULTS: Time invested in gaining tribal and health community engagement assisted in the development and design of clear protocols and processes for the study. Response rates were 57.6%, 48.3% and 57.2%. Co-operation rates (participation among those with whom contact was established) were 74.7%, 66.6% and 71.4%. CONCLUSIONS: Use of electoral rolls enables straightforward sampling but results in low response rates because electors have moved. Co operation rates highlight the acceptability of this research project to the participants; they indicate the strength of Kaupapa Maori Methodologies in engaging Maori participants and community. IMPLICATIONS: This study provides a model for conducting clinical/biomedical research projects that are compatible with cultural protocols and methodologies, in which the primary aim of the research was Maori health gain. PMID- 21627726 TI - Determinants of non-response in an occupational exposure and health survey in New Zealand. AB - OBJECTIVE: Study the determinants of non-response and the potential for non response bias in a New Zealand survey of occupational exposures and health. METHODS: A random sample of 10,000 New Zealanders aged 20-64 years were invited by mail to take part in a telephone survey. Multiple logistic regression was used to study the determinants of non-response. Whether occupational exposure, lifestyle and health indicators were associated with non-response was studied by standardising their prevalence towards the demographic distribution of the source population, and comparing early with late responders. RESULTS: The response rate was 37%. Younger age, Maori descent, highest and lowest deprivation groups and being a student, unemployed, or retired were determinants of non-contact. Refusal was associated with older age and being a housewife. Prevalence of key survey variables were unchanged after standardising to the demographic distribution of the source population. CONCLUSIONS: Following up the non-responders to the mailed invitations with telephone calls more than doubled the response rate and improved the representativeness of the sample. Although the response rate was low, we found no evidence of major non-response bias. IMPLICATIONS: Judgement regarding the validity of a survey should not be based on its response rate. PMID- 21627727 TI - Inpatient hospital use in the first year after release from prison: a Western Australian population-based record linkage study. AB - OBJECTIVE: To describe three aspects of inpatient use for ex-prisoners within the first 12 months of release from prison: the proportion of released prisoners who were hospitalised; the amount of resources used (bed days, separations and cost); and the most common reasons for hospitalisation. METHODS: Secondary analysis of whole-population linked prison and inpatient data from the Western Australian Data Linkage System. The main outcome measure was first inpatient admission within 12 months of release from prison between 2000 and 2002 and related resource use. RESULTS: One in five adults released from Western Australian prisons between 2000 and 2002 were hospitalised in the 12 months that followed, which translated into 12,074 inpatient bed days, 3,426 separations and costs of $10.4 million. Aboriginals, females and those released to freedom were most at risk of hospitalisation. Mental health disorders such as schizophrenia and depression, and injuries involving the head or face and/or fractures, accounted for 58.9% of all bed days. Ex-prisoners were 1.7 times more likely to be hospitalised during a year than Western Australia's general adult population of roughly the same age. CONCLUSIONS: Using whole-population administrative linked health and justice data, our findings show that prisoners are vulnerable to hospitalisation in the 12-month period following their release from prison, particularly Aboriginals, females and those with known mental health problems. IMPLICATIONS: Further research is needed to assess whether contemporary services to support community re-entry following incarceration have led to a measurable reduction in hospital contacts, especially for the subgroups identified in this study. PMID- 21627728 TI - Individual and household-level socioeconomic position is associated with harmful alcohol consumption behaviours among adults. AB - AIM: To examine associations between individual-, household- and neighbourhood level socioeconomic position (SEP) and harmful alcohol consumption. METHOD: Adults aged 18-76 residing in 50 neighbourhoods in Melbourne completed a postal questionnaire (n= 2349, 58.7% response rate). Alcohol-related behaviours were classified by risk of short- and long-term harm. Individual-, household- and neighbourhood-level SEP were ascertained by education, household income and proportion of low-income households, respectively. The association were examined by multi-level logistic regression. RESULTS: Participants lower education or household income were less likely to consume alcohol frequently compared to their more-advantaged counterparts. Lower-educated men were more likely to be at risk of short-term harm [OR 1.75 (1.23 - 2.48)]. Low-income women were less likely to be at risk of short-term harm [OR 0.44 (0.23 - 0.81)]. Neighbourhood disadvantage was not associated with alcohol consumption. CONCLUSION: Men and women from socioeconomically advantaged backgrounds were more frequent consumers of alcohol, whereas their disadvantaged counterparts drank less frequently but in greater quantities on each drinking occasion. IMPLICATIONS: Socioeconomic disadvantage at the individual and household levels may be an important determinant of alcohol consumption among Australian adults. PMID- 21627729 TI - The beneficial effects of preschool attendance on adult cardiovascular disease risk. AB - OBJECTIVE: To assess the effect of South Australian Kindergarten Union participation on adult cardiovascular behavioural risk factors. METHODS: Using a retrospective cohort design, this study examined the effect of attendance at a Kindergarten Union preschool from 1940 to 1972 on behavioural risk factors for cardiovascular disease in adults 34-67 years. Dichotomous outcomes were analysed using a generalised linear model (Poisson distribution) with robust variance estimates. Outcomes with more than two categories were analysed with a multinomial logistic model. RESULTS: There was a beneficial effect of preschool on high physical activity relative to sedentary and on ever smoking, but a negative effect on fruit consumption. Preschool attendance was not associated with alcohol risk or vegetable consumption under traditional criteria, however the point estimate for vegetable consumption was in the beneficial direction. The point estimates from the multinomial model suggested a step-wise decreasing risk for preschool attendees to have less risk of experiencing multiple behavioural risk factors (e.g. risk of five risk factors for preschool participants compared with non-participants). CONCLUSIONS AND IMPLICATIONS: Attendance at a Kindergarten Union preschool was associated with a reduced risk of two and an indication of benefit in a third behavioural risk factor in adulthood. This study provides some evidence for the potential health benefit of interventions outside of the health sector to prevent cardiovascular diseases, which are strongly associated with lifelong social disadvantage. PMID- 21627730 TI - Perceived weight status may contribute to education inequalities in five-year weight change among mid-aged women. AB - OBJECTIVES: To examine education differences in five-year weight change among mid aged adults, and to ascertain if this may be due to socioeconomic differences in perceived weight status or weight control behaviours (WCBs). METHODS: Data were used from the Australian Diabetes, Obesity and Lifestyle Study. Mid-aged men and women with measured weights at both baseline (1999-2000) and follow-up (2004 2005) were included. Percent weight change over the five-year interval was calculated and perceived weight status, WCBs and highest attained education were collected at baseline. RESULTS: Low-educated men and women were more likely to be obese at baseline compared to their high-educated counterparts. Women with a certificate-level education had a greater five-year weight gain than those with a bachelor degree or higher. Perceived weight status or WCBs did not differ by education among men and women, however participants that perceived themselves as very overweight had less weight gain than those perceiving themselves as underweight or normal weight. WCBs were not associated with five-year weight change. CONCLUSIONS AND IMPLICATIONS: The higher prevalence of overweight/obesity among low-educated women may be a consequence of greater weight gain in mid adulthood. Education inequalities in overweight/obesity among men and women made be due (in part) to overweight or obese individuals in low-educated groups not perceiving themselves as having a weight problem. PMID- 21627731 TI - Is greater variety of chocolates and confectionery in supermarkets associated with more consumption? PMID- 21627732 TI - A scientific audit of smartphone applications for the management of obesity. PMID- 21627733 TI - A primary healthcare clinic in a needle syringe program may contribute to HIV prevention by early detection of incident HIV in an injecting drug user. PMID- 21627734 TI - The impact of smoke-free laws on business revenue in hotels and licensed clubs in South Australia. PMID- 21627735 TI - Risk of lymph node metastasis in patients with pedunculated type early invasive colorectal cancer: a retrospective multicenter study. AB - Depth of invasion in early invasive colorectal cancer is considered an important predictive factor for lymph node metastasis. However, no large-scale reports have established the relationship between invasion depth of pedunculated type early invasive colorectal cancers and risk of lymph node metastasis. The aim of this retrospective cohort study was to clarify the risk of lymph node metastasis in pedunculated type early invasive colorectal cancers in a large series. Patients with pedunculated type early invasive colorectal cancer who underwent endoscopic or surgical resection at seven referral hospitals in Japan were enrolled. Haggitt's line was used as baseline and the invasion depth was classified into two groups, head invasion and stalk invasion. The incidence of lymph node metastasis was investigated between patients with head and stalk invasion. We analyzed 384 pedunculated type early invasive colorectal cancers in 384 patients. There were 154, 156, and 74 endoscopic resection cases, endoscopic resection followed by surgical operation, and surgical resection cases, respectively. There were 240 head invasion and 144 stalk invasion lesions. Among the lesions treated surgically, the overall incidence of lymph node metastasis was 3.5% (8/230). The incidence of lymph node metastasis was 0.0% (0/101) in patients with head invasion, as compared with 6.2% (8/129) in patients with stalk invasion. Pedunculated type early invasive colorectal cancers pathologically diagnosed as head invasion can be managed by endoscopic treatment alone. PMID- 21627736 TI - Marathon of eponyms: 19 Sjogren syndrome. AB - The use of eponyms has long been contentious, but many remain in common use, as discussed elsewhere (Editorial: Oral Diseases. 2009: 15; 185). The use of eponyms in diseases of the head and neck is found mainly in specialties dealing with medically compromised individuals (paediatric dentistry, special care dentistry, oral and maxillofacial medicine, oral and maxillofacial pathology, oral and maxillofacial radiology and oral and maxillofacial surgery) and particularly by hospital-centred practitioners. This series has selected some of the more recognized relevant eponymous conditions and presents them alphabetically. The information is based largely on data available from MEDLINE and a number of internet websites as noted below: the authors would welcome any corrections. This document summarizes data about Sjogren syndrome. PMID- 21627737 TI - Attitude and practice of the health care professionals towards the clinical practice guidelines in King Khalid University Hospital in Saudi Arabia. AB - RATIONALE, AIMS AND OBJECTIVES: To explore the opinion and practice of the health care providers in King Khalid University Hospital (KKUH) towards clinical practice guidelines (CPGs). METHODS: A cross-sectional self-reported anonymous survey was distributed to 2225 health care professionals working in KKUH clinical departments. RESULTS: The response rate was 56.5%. The respondents had a positive attitude towards CPGs; 90% thought that CPG unify patients' care and 96% agreed that CPGs improve the quality of services provided. The respondents' practice in using CPGs concurred with their attitude and opinion. A total of 86.3% agreed that CPGs changed the way they manage their patients and 71.8% agreed with the statement that they have already used CPGs in the management of the patients. Compared to nurses, physicians were significantly less likely to use CPGs in practice (P < 0.05); moreover, the practice of using guidelines differs significantly according to the years of experience, with 71% of respondents with experience of 15 years or more using CPGs in the management of their patients, compared to 60% among respondents with less years of experience (P < 0.05). CONCLUSION: The health care providers at KKUH have positive practice and attitude towards CPGs in general, which could positively influence the future introduction and implementation of evidence-based CPGs. PMID- 21627738 TI - Affective temperaments are associated with higher hopelessness and perceived disability in patients with open-angle glaucoma. AB - AIM: The aims of the study were to study: (i) affective temperaments in open angle glaucoma (OAG) patients with some degree of functional visual impairment; (ii) psychological well-being and perceived disability, and their associations with affective temperaments; and (iii) associations between visual impairment, affective temperaments and psychological well-being. METHOD: Participants were 91 outpatients (39 women, and 52 men) with open-angle glaucoma (OAG) who were assessed for Visual Field Index, Mean Defect and Pattern Standard Deviation. Patients were also administered the Beck Hopelessness Scale, the TEMPS-A (Rome), the Gotland Male Depression Scale, the Emotional Well-being Scale, the Perceived Disability Questionnaire and the Suicidal History Self-Rating Screening Scale. RESULTS: Open-angle glaucoma patients (compared with a non-clinical sample of university students) had higher scores on the TEMP-A dysthimic and hyperthimic traits and lower scores on cyclothimic, irritability and anxiety traits. Such temperament variability was not linked to differences in severity of glaucoma. We did not find strong evidence supporting the fact that measures of visual impairment were linked to emotional well-being and depression. However, logistic regression analysis revealed that patients may have different patterns related to their illness according to specific temperaments. CONCLUSION: Patients with OAG may have different temperament profiles than non-clinical individuals. Such categorisation may be useful for predicting how they face the illness, for providing better care as well as for early recognition of mood disorders symptoms. PMID- 21627739 TI - Intensive care unit experience of haemopoietic stem cell transplant patients. AB - BACKGROUND: Previous research at our institution (1988-1998) established an intensive care unit (ICU) and hospital mortality between 70% and 80% in haemopoietic stem cell transplant (HSCT) patients requiring ICU admission. AIMS: This study explored mortality in a more contemporary cohort while comparing outcomes to published literature and our previous experience. METHODS: Retrospective chart review of HSCT patients admitted to ICU between December 1998 and June 2008. RESULTS: Of 146 admissions, 53% were male, with a mean age of 44 years, an Acute Physiologic and Chronic Health Evaluation II score of 28 and Sepsis Organ Failure Assessment score of 11. Fifty-six per cent had graft versus host disease (GVHD), with respiratory failure (67%) being the most common admission diagnosis. All but one received mechanical ventilation. The ICU and hospital mortality were 42% (72% 1988-1998 cohort) and 64% (82% 1998-1998 cohort) respectively. The 6- and 12-month survivals were 29% and 24% respectively for the 1998-2008 cohort. Dying in ICU was independently predicted by fungal infection (P= 0.02) and early onset of organ failure (P < 0.001), while GVHD (P= 0.04) predicted survival. Mortality at 12 months was independently predicted by the acute physiology score (P= 0.002), increasing number of organ failures (P= 0.001), and cytomegalovirus positive serology (P= 0.005), while blood stream infection (P= 0.003), an antibiotic change on admission to the ICU (P= 0.007) and a diagnosis of non-Hodgkin lymphoma (P= 0.02) predicted survival. CONCLUSION: Our study found that acute admission of HSCT patients to the ICU is associated with improved survival compared to our previous experience, with organ failure progression a strong predictor of ICU outcome, and specific disease characteristics contributing to long-term survival. PMID- 21627740 TI - Suboptimal management of cardiovascular risk factors in coronary heart disease patients in primary care occurs particularly in women. AB - BACKGROUND: Patients with established coronary heart disease (CHD) are at the highest risk of further events. Despite proven therapies, secondary prevention is often suboptimal. General practitioners (GPs) are in an ideal position to improve secondary prevention. AIM: To contrast management of cardiovascular risk factors in patients with established CHD in primary care to those in clinical guidelines and according to gender. METHODS: GPs throughout Australia were approached to participate in a programme incorporating a disease management software (mdCare) program. Participating practitioners (1258 GPs) recruited individual patients whose cardiovascular risk factor levels were measured. RESULTS: The mdCare programme included 12,509 patients (58% male) diagnosed with CHD. Their mean age was 71.7years (intra-quartile range 66-78) for men and 74years (intra-quartile range 68-80) for women. Low-density-lipoprotein cholesterol was above target levels in 69% (2032) of women compared with 58% (2487) in men (P < 0.0001). There was also a higher proportion of women with total cholesterol above target levels (76%, 3592) compared with men (57%, 3787) (P < 0.0001). In patients who were prescribed lipid-lowering medication, 53% (2504) of men and 72% (2285) of women continued to have a total cholesterol higher than recommended target levels (P < 0.0001). Overall, over half (52%, 6538) had at least five cardiovascular risk factors (55% (2914) in women and 50% (3624) in men, P < 0.0001). CONCLUSION: This study found less intensive management of cardiovascular risk factors in CHD patients, particularly among women, despite equivalent cardiovascular risk. This study has shown that these patients have multiple risk factors where gender also plays a role. PMID- 21627741 TI - Gastro-oesophageal reflux and respiratory symptoms in Busselton adults: the effects of bodyweight and sleep apnoea. AB - BACKGROUND/AIM: Respiratory symptoms and gastro-oesophageal reflux disease (GORD) are common within the general population. Although a number of epidemiological studies have addressed their relationship, none has investigated the confounding effects of body mass index (BMI) and obstructive sleep apnoea (OSA), both of which are associated with reflux. METHODS: Men and women (2700) from the 2005 2007 cross-sectional Busselton health survey were included. Questionnaire data included demography, information on general health, asthma, cough, wheeze, dyspnoea and reflux symptoms (never, monthly or less often and weekly or more often). BMI, risk of OSA (Berlin questionnaire definition), spirometry and airway hyperresponsiveness (AHR) were recorded. The effects of BMI and OSA on the relationship between respiratory and reflux symptoms were examined using logistic regression models, expressed as adjusted odds ratios for risk of respiratory symptoms by reflux symptom category. RESULTS: Fifty per cent had reflux symptoms (5-10% weekly or more often). Reflux symptoms had strong positive, dose-related associations with cough/phlegm, breathlessness, chest tightness and wheeze in the last 12 months (P < 0.001), but were not related to diagnosed asthma or AHR. Twenty-three per cent were at high risk of OSA and 63% had a BMI of >25 (22% > 30). Increased weight or high risk of OSA did not affect the relationship between respiratory symptoms and reflux symptoms. CONCLUSION: The relationship between reflux and respiratory symptoms was independent of BMI, high risk of OSA or AHR. These findings suggest that reflux contributes directly to respiratory symptoms. PMID- 21627742 TI - Management of heart conditions in older rural and urban Australian women. AB - BACKGROUND: Cardiovascular disease death rates are higher among Australians from regional and remote areas than those from major cities. AIMS: To investigate the causes of excess mortality from heart disease in rural compared to urban Australian women aged over 75 years. METHODS: In 2004, 944 older urban and rural participants in the Australian Longitudinal Study on Women's Health aged 77 83 years with self-reported ischaemic heart disease (IHD), heart failure or atrial fibrillation took part in a nested cross-sectional substudy. We used clinical guidelines to determine key management issues for these conditions. Using logistic regression we calculated odds ratios (OR) and 95% confidence intervals (CI) to assess the relationship between management and area of residence. RESULTS: These older Australian women often did not receive recommended management for their heart conditions. Only 30% reported having had an echocardiogram. Reported use of statins and beta-blockers was low among women with IHD (58% and 41% respectively) and only 32% of women reporting heart failure were taking angiotensin-converting enzyme inhibitors. Women from regional/remote areas had greater odds of reporting never having seen a cardiologist (OR = 3.88, 95% CI 1.72-8.72) and never having had an echocardiogram than women from major cities (OR = 2.86, 95% CI 1.42-5.75). Medication use was similar for rural and urban women. CONCLUSIONS: Our results suggest that best-practice treatments for heart conditions are suboptimally provided to older women. In addition, they suggest differential use of some health services, which might help explain higher cardiovascular mortality among rural compared with urban women. PMID- 21627743 TI - Medical oncology clinics through videoconferencing: an acceptable telehealth model for rural patients and health workers. AB - BACKGROUND: Since 2007, Townsville Cancer Centre (Queensland, Australia) has provided routine and urgent medical oncology services to rural and remote communities through videoconferencing. At remote sites, patients were accompanied by doctors and other health workers. The aim of this study was to describe satisfaction of patients and rural health workers with this model of teleoncology. METHODS: Between May 2007 and June 2010, 55 videoconferencing patients were invited to participate in a questionnaire-based telephone survey after informed consent. The survey included responses to 16 satisfaction statements using a 5-point Likert scale. Perspectives of health workers involved were recorded using open-ended questions on six themes related to telehealth. RESULTS: Among the 50 participating cancer patients, median age was 56 years (range 28-83). Seventy-six per cent of patients thought specialist physical examination was important despite local doctors performing it. Seventy-six per cent of patients felt the presence of accompanying local health workers was not important. Seventy-eight per cent of patients preferred to be seen in Mt Isa for the first consultation through videoconferencing than travelling to Townsville. More than 80% of patients were in agreement with the rest of the satisfaction statements. Ninety-six per cent of patients were in agreement with the question relating to overall satisfaction. Eighteen health workers participated and their responses and attitudes were favourable towards telehealth. CONCLUSION: Satisfaction with this model of care was proven to be high among both patients and health workers. Benefits perceived included effective communication between patients and specialists, reduced travel time and money expenditure, and superior specialist support for rural health workers. PMID- 21627744 TI - Monocyte chemoattractant protein-1 is associated with silent cerebral infarction in patients on haemodialysis. AB - BACKGROUND: In patients with chronic renal failure undergoing haemodialysis (HD), silent cerebral infarctions (SCI) are associated with high mortality. Levels of monocyte chemoattractant protein-1 (MCP-1) increase with renal dysfunction and may be a novel predictor for cerebrovascular events. We tested the hypothesis that increased MCP-1 concentration correlate with the occurrence of SCI in HD patients. METHODS: Using cranial magnetic resonance imaging (MRI) findings, 52 Japanese patients undergoing HD were divided into two groups: with SCI (61 +/- 7 years, mean +/- SD, n= 28) and without SCI (60 +/- 6 years, n= 24). The gender, metabolic profiles and MCP-1 concentration were compared between the two groups. RESULTS: The level of MCP-1 was higher in the with-SCI group than in the without SCI group (P < 0.0001). The proportion of smokers was higher in the with-SCI group (P < 0.05) than in the without-SCI group. Plasma level of high-density lipoprotein cholesterol was lower, while uric acid level was higher, in the with SCI group (P < 0.05 and P < 0.05 respectively) compared to the without-SCI group. Multiple logistic regression analysis identified MCP-1 level as being significantly associated with the presence of SCI (odds ratio 1.48, 95% confidence interval = 1.10-5.75, P < 0.0001). CONCLUSIONS: This study indicates that patients with chronic renal failure who are maintained on HD exhibit an increased prevalence of SCI, and that MCP-1 is significantly associated with the presence of SCI in HD patients. PMID- 21627745 TI - Performance of comorbidity indices in measuring outcomes after acute myocardial infarction in Australian indigenous and non-indigenous patients. AB - BACKGROUND: Indigenous Australians have higher prevalence of chronic diseases and worse acute care outcomes than other Australians. The extent to which higher chronic disease comorbidity levels are responsible for their worse outcomes is not clear, and the performance of comorbidity indices has not been assessed for this population with very high comorbidity levels. METHODS: Using hospital separations data, the Charlson and Elixhauser comorbidity indices were used to measure chronic disease prevalence in 2035 indigenous and non-indigenous patients hospitalised after their first acute myocardial infarction (AMI) in the Northern Territory of Australia between 1992 and 2004, and to adjust for comorbidity in multivariate analysis of mortality outcomes (in-hospital and long-term deaths from coronary heart disease and all causes). Index performance was assessed by the difference between C statistic, Akaike information criterion statistic and estimate of excess indigenous mortality in models with and without comorbidity adjustment. RESULTS: Comorbidity index scores were higher for indigenous than non indigenous patients and increased considerably over time, at least partly because of information bias. Indigenous patients' higher risk of in-hospital all-cause death was almost fully explained by their higher comorbidity levels. Their higher risk of long-term coronary heart disease and all-cause death was partially explained by higher comorbidity levels. Charlson and Elixhauser indices performed satisfactorily and similarly in this population. CONCLUSION: Comorbidity indices performed well in a population with very high chronic disease prevalence. After adjusting for comorbidity, short-term outcomes were similar for indigenous and non-indigenous AMI patients, but comorbidity at the time of the acute episode only partly explained the worse long-term outcomes for indigenous patients. PMID- 21627746 TI - Management of metastatic renal cell carcinoma in the era of targeted therapies. AB - BACKGROUND: Metastatic renal cell cancer is associated with poor prognosis and survival and is resistant to conventional chemotherapy. Therapeutic targeting of molecular pathways for tumour angiogenesis and other specific activation mechanisms offers improved tumour response and prolonged survival. AIMS: To conduct a retrospective audit of metastatic renal cell carcinoma patients treated with targeted therapies. METHODS: Data were extracted from clinical records of patients undergoing targeted treatment between 2005 and 2009 at two hospital sites. Data collected included pathology, systemic therapy class, toxicity and survival. Univariate and multivariate survival analyses were performed. RESULTS: Sixty-one patients were treated with 102 lines of therapy with a median overall survival (OS) of 23 months, median time to failure of first-line treatment (TTF1) of 10 months and median time to failure of second-line treatment (TTF2) of 5.2 months. Time from first diagnosis to treatment >12 months was significantly associated with improved OS, longer TTF1, TTF2 and response to first-line anti vascular endothelial growth factor receptor tyrosine kinase inhibitors (anti-VEGF TKI) therapy. Variables associated with tumour biology, natural history and the systemic inflammatory response were associated with improved OS and TTF1. Development of hypertension was predictive of anti-VEGF TKI outcome. Toxicities were as expected for each drug class. CONCLUSIONS: Survival and toxicity outcomes from two Australian sites are comparable to published data. The adverse event profile differs to conventional chemotherapy. Clinicians caring for patients with metastatic renal cancer will need to become familiar with these toxicities and their management as these agents enter widespread use. PMID- 21627747 TI - Identifying patient-specific beliefs and behaviours for conversations about adherence in asthma. AB - BACKGROUND: Asthma guidelines advise addressing adherence at every visit, but no simple tools exist to assist clinicians in identifying key adherence-related beliefs or behaviours for individual patients. AIMS: To identify potentially modifiable beliefs and behaviours that predict electronically recorded adherence with controller therapy. METHODS: Patients aged >= 14 years with doctor-diagnosed asthma who were prescribed inhaled corticosteroid/long-acting beta(2)-agonist (ICS/LABA) completed questionnaires on medication beliefs/behaviours, side effects, Morisky adherence behaviour score and Asthma Control Test (ACT), and recorded spirometry. Adherence with ICS/LABA was measured electronically over 8 weeks. Predictors of adherence were identified by univariate and multivariate analyses. RESULTS: 99/100 patients completed the study (57 female; forced expiratory volume in 1 s mean +/- standard deviation 83 +/- 23% predicted; ACT 19.9 +/- 3.8). Mean electronically recorded adherence (n= 85) was 75% +/- 25, and mean self-reported adherence was 85% +/- 26%. Factor analysis of questionnaire items significantly associated with poor adherence identified seven themes: perceived necessity, safety concerns, acceptance of asthma chronicity/medication effectiveness, advice from friends/family, motivation/routine, ease of use and satisfaction with asthma management. Morisky score was moderately associated with actual adherence (r=-0.45, P < 0.0001). In regression analysis, 10 items independently predicted adherence (adjusted R(2) = 0.67; P < 0.001). Opinions of friends/family about the patient's medication use were strongly associated with poor adherence. Global concerns about ICS/LABA therapy were more predictive of poor adherence than were specific side-effects; the one-third of patients who reported experiencing side-effects from their steroid inhaler had lower adherence than others (mean 62% vs 81%; P= 0.015). CONCLUSIONS: This study identified several specific beliefs and behaviours which clinicians could use for initiating patient-centred conversations about medication adherence in asthma. PMID- 21627748 TI - Cross-ecosystem differences in stability and the principle of energy flux. AB - Here, we review consumer-resource (C-R) theory to show that the paradox of enrichment is a special case of a more general theoretical result. That is, we show that increased energy flux, relative to the consumer loss rate, makes C-R interactions top heavy (i.e., greater C:R biomass ratio) and less stable. We then review the literature on the attributes of aquatic and terrestrial ecosystems to argue that empirical estimates of parameters governing energy flux find that aquatic ecosystems have higher rates of relative energy flux than terrestrial ecosystems. Consistent with theory, we then review empirical work that shows aquatic ecosystems have greater herbivore:plant biomass ratios while we produce novel data to show that aquatic ecosystems have greater variability in population dynamics than their terrestrial counterparts. We end by arguing that theory, allometric relationships and a significant, negative correlation between body size and population variability suggest that these results may be driven by the smaller average body sizes of aquatic organisms relative to terrestrial organisms. PMID- 21627749 TI - Global histone H4K20 trimethylation predicts cancer-specific survival in patients with muscle-invasive bladder cancer. AB - OBJECTIVE: *To determine the role of global histone methylation as a prognostic parameter in patients with bladder cancer. PATIENTS AND METHODS: *We used a tissue microarray with samples from patients with non-muscle-invasive bladder cancer (NMIBC; n= 161), muscle-invasive bladder cancer (MIBC, n= 127), normal urothelium (NU; n= 31) and bladder cancer metastases (METS; n= 31) to determine global histone methylation (me) levels at histone H3 lysine 4 (H3K4) and H4K20. RESULTS: *Global histone modification levels (H3K4me1, H3K4me3, H4K20me1, H4K20me2, and H4K20me3) were lower in bladder cancer samples than in NU tissue *Global levels of H3K4me1, H4K20me1, H4K20me2 and H4K20me3 were decreasing from NU over NMIBC and MIBC to METS. *H4K20me1 levels were increased in patients with NMIBC with advanced pTstage and less differentiated bladder cancer. *In patients with MIBC, pTstage was negatively correlated with H3K4me1, H4K20me1 and H4K20me2 levels. *H4K20me3 levels were significantly correlated in a univariate and multivariate model with bladder cancer-specific mortality after radical cystectomy in patients with MIBC. CONCLUSION: *Global histone methylation levels may help to identify patients with bladder cancer with poor prognosis after radical cystectomy. PMID- 21627750 TI - Optimizing prostate cancer detection during biopsy by standardizing the amount of tissue examined per core. AB - OBJECTIVE: To investigate the effect on cancer detection by varying the number of cores taken for prostate biopsy according to the size of the prostate. PATIENTS AND METHODS: A retrospective review of a prospectively registered prostate biopsy database identified 3040 consecutive patients undergoing prostate biopsy at a Veterans Administration Hospital between 1994 and 2008. Of 2224 biopsies, 681 (31%) were found to have cancer and 1540 (69%) had negative biopsies. Prostate volume to biopsy core ratios (volume/number of cores) were derived and a comparative analysis was performed to determine the impact on cancer detection rates. RESULTS: The median prostate volume was significantly smaller for those patients diagnosed with prostate cancer than for those with negative biopsies (33 vs 43 cc, P= 0.01). The median number of cores was the same for both groups of patients (median 12, P= 0.66). The median transrectal ultrasonography TRUS size/core ratio was 3.5 [interquartile range (IQR) 2.5] for patients with identified cancer as compared with 4.7 (IQR = 3.9) for those with negative biopsies (P= 0.000). On multivariable logistic regression analysis TRUS size/core ratio had a significant impact on cancer detection with a relative risk ratio of 1.29 (95% confidence interval, 1.1-1.5, P= 0.001) even when controlled for age, race, prostate volume, digital rectal examination and prostate-specific antigen level. CONCLUSIONS: Prostate cancer detection can be enhanced by individualizing the number of cores performed to a real-time prostate volume sampling. The present study emphasizes that optimal cancer detection rates were observed when a ratio of 3.5 cc per tissue core was achieved. Proper prospectively designed studies must be performed to further validate these findings. PMID- 21627751 TI - The clinical features and management of testicular germ cell tumours in patients aged 60 years and older. AB - What's known on the subject ? and What does the study add? The treatment of younger men with testicular germ cell cancers is well documented with established intensive chemotherapy regimens for those with advanced disease. Although the majority of patients present in the third or fourth decade, men also present in later life. These patients are typically excluded from clinical trials and there are no contemporary published series describing their management. This series describes the management of older patients with testicular germ cell tumours at both early and advanced stages of disease. Patients with stage I seminoma can be safely managed with all recognised treatment strategies and state I non-seminomas were managed with surveillance. Cure can still be achieved in older patients with advance germ cell tumours however chemotherapy regimens developed in younger patients must be tailored to the presence of co-morbidity. OBJECTIVES: * To review the practice of a large referral centre for the management of older patients with testicular germ cell cancer (GCC). * There are few published data available on the management of testicular GCC in elderly patients, who often have medical comorbidities and have been excluded from clinical trials. PATIENTS AND METHODS: * We reviewed our prospectively collected database for patients presenting with GCC who were aged >=60 years. * Details of presentation, management and outcome were recorded. RESULTS: * In total, 60 patients aged >=60 years were identified from 1461 patients treated with GCC from 1979-2005, representing 4% of the total population. * Median age was 67 years, 44 had seminoma (73%) and 16 had non-seminoma histology (27%). * Stage I seminoma patients were managed with surveillance, adjuvant radiotherapy and adjuvant carboplatin. All stage I non-seminomas underwent surveillance. * In total, 15 patients received systemic chemotherapy for metastatic disease with modified bleomycin, etoposide and cisplatin; etoposide and cisplatin; carboplatin-based regimens; or other combinations. Toxicity was manageable, with no toxic deaths. * In total, four patients (6.7%) died of GCC. CONCLUSIONS: * In elderly patients, GCC should be managed with curative intent. * Conventional therapies are tolerable for most men with stage I seminoma. In metastatic disease, comorbidity may necessitate treatment modifications. * Most patients are cured with manageable toxicity. PMID- 21627752 TI - Evaluation of the safety and efficiency of the dorsal slit and sleeve methods of male circumcision provided by physicians and clinical officers in Rakai, Uganda. AB - OBJECTIVE: To assess the safety and efficiency of the dorsal slit and sleeve male circumcision (MC) procedures performed by physicians and clinical officers (COs). PATIENTS AND METHODS: We evaluated the time required for the MC procedure (efficiency) and moderate/severe adverse events (AEs) for MC (safety) by trained physicians and COs using the sleeve and dorsal slit MC methods in a service programme. Univariate and multiple regressions with robust variance estimation were used to assess factors associated with operative duration (linear) and AEs (logistic). RESULTS: Six physicians and eight COs conducted 1934 and 3218 MCs, respectively; there were 2471 dorsal slit and 2681 sleeve MC procedures. The overall mean operative duration was 33 min for newly trained providers, which decreased to ~20 min after ~100 MCs. The adjusted mean operative duration for dorsal slit MC was significantly shorter than that for the sleeve MC method (Delta - 2.7 min, P < 0.001). The operative duration was longer for COs than physicians for the sleeve procedure, but not the dorsal slit procedure; however this difference reduced with increasing numbers of MCs completed. The unadjusted AE rates were 0.6% for dorsal slit MC and 1.4% for the sleeve method (P = 0.006) and 1.5% for physicians and 0.68% for COs (P = 0.003); however, there were no significant differences after multivariate adjustment. Use of bipolar cautery significantly reduced operative duration (Delta - 4.0 min, P = 0.008), but was associated with higher AE rates (adjusted odds ratio 2.13, 95% confidence interval 1.26-3.61, P = 0.005). CONCLUSION: The dorsal slit MC method is faster than sleeve resection, and can be safely performed by non-physicians; however, use of bipolar cautery may be inadvisable in this setting. PMID- 21627753 TI - Determination of cardiac output by ultrasound velocity dilution in normovolemia and hypovolemia in dogs. AB - OBJECTIVE: The purpose of this study was to compare CO measured by use of lithium dilution (LiDCO) and ultrasound velocity dilution (UDCO) in a canine model of acute hemorrhage. STUDY DESIGN: Prospective descriptive evaluation. ANIMALS: Twelve dogs (15-34 kg). METHODS: Dogs were anesthetized and instrumented to measure direct blood pressure, heart rate, arterial blood gases and CO. The CO was measured by use of LiDCO and UDCO techniques. Measurements were obtained from each animal at baseline and during a low CO state (hemorrhagic state). Measurements were converted to cardiac index (CI = CO/BSA) values for statistical analysis. To measure CO using UDCO, a 20 mL bolus of 0.9% sodium chloride was administered and CO was calculated from the transient dilution of blood proteins created by the injection bolus. Hypovolemia was induced by withdrawing 40% of the blood volume until the mean arterial blood pressure (MAP) was stable at 40 mmHg for 10 minutes. Agreement was determined using Bland & Altman analysis and concordance correlation coefficients. RESULTS: Twenty-four comparisons were made. Lithium determinations of CI ranged between 7.5 and 1.3 L minute(-1) m(-2), and the mean overall difference between the two methods was -0.40 L minute(-1) m(-2). The mean relative bias was -17 +/- 21% (limits of agreements: -59% to 25%). There was no significant effect of state of CI on bias or relative bias (p = 0.24 and p = 0.10, respectively). The concordance correlation coefficient between LiDCO and UDCO as 0.88 (p < 0.0001). CONCLUSIONS: When compared to lithium dilution, the UDCO technique is a viable method for measuring cardiac output in a model of normovolemia and hypovolemia in dogs. PMID- 21627754 TI - The effect of ketamine on the MACBAR of sevoflurane in dogs. AB - OBJECTIVE: To determine the effect of intravenous ketamine on the minimum alveolar concentration of sevoflurane needed to block autonomic response (MAC(BAR)) to a noxious stimulus in dogs. STUDY DESIGN: Randomized, crossover, prospective design. ANIMALS: Eight, healthy, adult male, mixed-breed dogs, weighing 11.2-16.1 kg. METHODS: Dogs were anesthetized with sevoflurane on two occasions, 1 week apart, and baseline MAC(BAR) (B-MAC(BAR)) was determined on each occasion. MAC(BAR) was defined as the mean of the end-tidal sevoflurane concentrations that prevented and allowed an increase (>=15%) in heart rate or invasive mean arterial pressure in response to a noxious electrical stimulus (50 V, 50 Hz, 10 ms). Dogs then randomly received either a low-dose (LDS) or high dose series (HDS) of ketamine, and treatment MAC(BAR) (T-MAC(BAR)) was determined. The LDS had an initial loading dose (LD) of 0.5 mg kg(-1) and constant rate infusion (CRI) at 6.25 MUg kg(-1) minute(-1), followed, after T MAC(BAR) determination, by a second LD (1 mg kg(-1)) and CRI (12.5 MUg kg(-1) minute(-1)). The HDS had an initial LD (2 mg kg(-1)) and CRI (25 MUg kg(-1) minute(-1)) followed by a second LD (3 mg kg(-1)) and CRI (50 MUg kg(-1) minute( 1)). Data were analyzed with a mixed-model anova and are presented as LSM +/- SEM. RESULTS: The B-MAC(BAR) was not significantly different between treatments. Ketamine at 12.5, 25, and 50 MUg kg(-1) minute(-1) decreased sevoflurane MAC(BAR), and the maximal decrease (22%) occurred at 12.5 MUg kg(-1) minute(-1). The percentage change in MAC(BAR) was not correlated with either the log plasma ketamine or norketamine concentration. CONCLUSIONS AND CLINICAL RELEVANCE: Ketamine at clinically relevant doses of 12.5, 25, and 50 MUg kg(-1) minute(-1) decreased sevoflurane MAC(BAR), although the reduction was neither dose-dependent nor linear. PMID- 21627755 TI - The use of different doses of metamizol for post-operative analgesia in dogs. AB - OBJECTIVE: To evaluate the post-operative analgesic effect of metamizol (dipyrone) administered intravenously at three different doses (15 mg kg(-1), 25 mg kg(-1) and 35 mg kg(-1)) compared to placebo in dogs undergoing ovariohysterectomy. STUDY DESIGN: Prospective, comparative, randomized, blinded trial. ANIMALS: Forty healthy bitches, aged 1-6 years, weighing 10-35 kg METHODS: The animals were randomly divided into four groups and received their respective treatments immediately after surgery: placebo group (0.9% saline solution), D15 group (metamizol 15 mg kg(-1) IV), D25 group (metamizol 25 mg kg(-1) IV), D35 group (metamizol 35 mg kg(-1) IV). The following variables were measured: sedation, pulse rate (PR), respiratory rate (f(R)), arterial blood pressure (ABP), plasma catecholamines, serum cortisol, blood urea nitrogen (BUN) and creatinine metabolites, albumin, alanine aminotransferase (ALT), alkaline phosphatase (ALP), hemogram, platelet counts and level of analgesia which was assessed by visual analog (VAS), descriptive and behavioral scales. Patients were monitored for 48 hours after the administration of the analgesic agent. Rescue analgesia (tramadol, 2 mg kg(-1), intramuscularly) was provided for animals with pain scores >=4, as determined by the VAS or descriptive scale. RESULTS: The D25 and D35 groups showed equivalent post-operative analgesia, as shown by decreased pain scores, according to the three different pain scales, and fewer animals that required rescue analgesia. Significantly lower serum cortisol concentrations were observed in the D25 and D35 groups when compared to the placebo and D15 groups. No hematologic, renal, hepatic or clinical adverse effects were observed during the treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Metamizol administered intravenously at 25 or 35 mg kg(-1) can provide adequate post-operative analgesia in bitches undergoing ovariohysterectomy. PMID- 21627756 TI - Determination of the sevoflurane sparing effect of methadone in cats. AB - OBJECTIVE: To determine the magnitude and duration of sevoflurane minimum alveolar concentration (MAC) reduction following a single intravenous (IV) dose of methadone in cats. STUDY DESIGN: Prospective experimental study. ANIMALS: Eight (four females and four males) healthy mixed-breed adult (1-2 years) cats weighing 5.82 +/- 0.42 kg. METHODS: Anesthesia was induced and maintained with sevoflurane. Intravenous catheters facilitated administration of methadone and lactated Ringer's solution. After baseline MAC determination in triplicate using a tail clamp technique, 0.3 mg kg(-1) of methadone was administered IV. End-tidal sevoflurane concentration (e'SEVO) was reduced and MAC was redetermined. In an effort to determine the duration of MAC reduction, measurements were repeated in a stepwise manner until MAC values returned to baseline. After the last stimulation, the e'SEVO was increased to 1.2 individual MAC for 15 minutes, then sevoflurane was discontinued and cats were allowed to recover from anesthesia. RESULTS: Baseline sevoflurane MAC was 3.18 +/- 0.06%. When compared with baseline the sevoflurane MAC after methadone administration was significantly reduced by 25, 15 and 7% at 26, 76 and 122 minutes, respectively. The final MAC value (3.09 +/- 0.07%) determined 156 minutes after methadone administration was not significantly different from baseline. CONCLUSIONS AND CLINICAL RELEVANCE: Intravenous methadone (0.3 mg kg(-1)) significantly decreased MAC of sevoflurane in cats but the effect was short-lived. PMID- 21627757 TI - Twitch potentiation: a potential source of error during neuromuscular monitoring with acceleromyography in anesthetized dogs. AB - OBJECTIVE: To measure twitch potentiation (the staircase phenomenon) in anesthetized dogs, and assess its relevance during neuromuscular monitoring with acceleromyography (AMG). STUDY DESIGN: Randomized, prospective clinical trial. ANIMALS: Sixteen dogs undergoing ovariohysterectomy. METHODS: Under isoflurane anesthesia, neuromuscular function was monitored with train-of-four (TOF) stimuli every 15 seconds and quantified by AMG. Neuromuscular blockade (NMB) was produced with 0.15 mg kg(-1) atracurium IV. Dogs were randomly divided into two groups; a potentiation group (PG) in which TOF stimulation was applied for 20 minutes before atracurium was administered; and a control group (CG) where no such time was allowed. In both groups, the AMG was calibrated (at tCAL) just before atracurium was administered. TOF stimulation continued throughout the experiment in all dogs. The height of the first twitch (T(1)) (expressed as a fraction of T(1) at tCAL) and train-of-four ratio (TOFR) were recorded until TOFR returned to >=90%. RESULTS: In PG, T(1) increased significantly (p = 0.0078) from a median of 102% (range, 95, 109) at baseline to 118% (100, 142) at 20 minutes. In PG, no difference was found between T(1) at tCAL (immediately before atracurium administration) and T(1) when neuromuscular transmission returned (p = 0.42). In the CG, T(1) increased significantly between tCAL and the time neuromuscular transmission returned (p = 0.027). TOFR did not increase during twitch potentiation (all p = 0.32). CONCLUSIONS AND CLINICAL RELEVANCE: T(1) increased significantly during 20 minutes of uninterrupted TOF stimulation in the absence of NMB, establishing that twitch potentiation occurs in anesthetized dogs. With no time for potentiation, T(1) increased during the course of recovery from NMB; this phenomenon introduces a bias in T(1) measurements and could affect studies reporting potency and duration of NMB based on T(1) or single twitches. TOFR was unaltered by potentiation emphasizing its clinical usefulness for excluding post operative residual NMB. PMID- 21627758 TI - The effects of lactated Ringer's solution (LRS) or LRS and 6% hetastarch on the colloid osmotic pressure, total protein and osmolality in healthy horses under general anesthesia. AB - OBJECTIVE: To investigate changes in colloid osmotic pressure (COP), total protein (TP) and osmolality (OSM) during anesthesia in horses given intravenous lactated Ringer's solution (LRS) or LRS and hetastarch (HES). STUDY DESIGN: Prospective, clinical trial. ANIMALS: Fourteen horses presented for surgery. Mean age 8.3 +/- 1.9 years; mean weight 452 +/- 25 kg. METHODS: Horses were premedicated with xylazine intravenously (IV); anesthesia was induced with ketamine and diazepam IV, and maintained with sevoflurane. Butorphanol was administered IV with pre-medications or immediately after induction. Xylazine was administered IV for recovery if necessary. LRS was administered IV to all horses with a target rate of 5-10 mL kg(-1) hour(-1). Half of the horses also received 6% HES, 2.5 mL kg(-1) over 1 hour in addition to LRS. Horses that received LRS only were considered the LRS group. Horses that received both LRS and HES were considered the LRS/HES group. Blood was drawn pre- and post-anesthesia, immediately following induction, and every 30 minutes throughout anesthesia. COP, TP and OSM were measured. RESULTS: COP and TP significantly decreased at similar rates for both treatment groups from pre-anesthetic values. Pre-anesthetic COP was significantly greater in the LRS group when compared to the LRS/HES group pre , post- and throughout anesthesia. In the LRS group post-anesthetic OSM was significantly different than the pre-anesthesia value and that for the LRS/HES group. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of IV HES (2.5 mL kg( 1), over 1 hour) in combination with LRS does not attenuate the decrease in COP typically seen during anesthesia with crystalloid administration alone. Based on these results, administration of HES at this rate and total volume would not be expected to prevent fluid shifts into the interstitium through its effects on COP. PMID- 21627759 TI - Manipulating corn germplasm to increase recombinant protein accumulation. AB - Using plants as biofactories for industrial enzymes is a developing technology. The application of this technology to plant biomass conversion for biofuels and biobased products has potential for significantly lowering the cost of these products because of lower enzyme production costs. However, the concentration of the enzymes in plant tissue must be high to realize this goal. We describe the enhancement of the accumulation of cellulases in transgenic maize seed as a part of the process to lower the cost of these dominant enzymes for the bioconversion process. We have used breeding to move these genes into elite and high oil germplasm to enhance protein accumulation in grain. We have also explored processing of the grain to isolate the germ, which preferentially contains the enzymes, to further enhance recovery of enzyme on a dry weight basis of raw materials. The enzymes are active on microcrystalline cellulose to release glucose and cellobiose. PMID- 21627760 TI - Transcript-specific, single-nucleotide polymorphism discovery and linkage analysis in hexaploid bread wheat (Triticum aestivum L.). AB - Food security is a global concern and substantial yield increases in cereal crops are required to feed the growing world population. Wheat is one of the three most important crops for human and livestock feed. However, the complexity of the genome coupled with a decline in genetic diversity within modern elite cultivars has hindered the application of marker-assisted selection (MAS) in breeding programmes. A crucial step in the successful application of MAS in breeding programmes is the development of cheap and easy to use molecular markers, such as single-nucleotide polymorphisms. To mine selected elite wheat germplasm for intervarietal single-nucleotide polymorphisms, we have used expressed sequence tags derived from public sequencing programmes and next-generation sequencing of normalized wheat complementary DNA libraries, in combination with a novel sequence alignment and assembly approach. Here, we describe the development and validation of a panel of 1114 single-nucleotide polymorphisms in hexaploid bread wheat using competitive allele-specific polymerase chain reaction genotyping technology. We report the genotyping results of these markers on 23 wheat varieties, selected to represent a broad cross-section of wheat germplasm including a number of elite UK varieties. Finally, we show that, using relatively simple technology, it is possible to rapidly generate a linkage map containing several hundred single-nucleotide polymorphism markers in the doubled haploid mapping population of Avalon * Cadenza. PMID- 21627761 TI - Did sulfate availability facilitate the evolutionary expansion of chlorophyll a+c phytoplankton in the oceans? AB - During the Mesozoic Era, dinoflagellates, coccolithophorids and diatoms became prominent primary producers in the oceans, succeeding an earlier biota in which green algae and cyanobacteria had been proportionally more abundant. This transition occurred during an interval marked by increased sulfate concentration in seawater. To test whether increasing sulfate availability facilitated the evolutionary transition in marine phytoplankton, the cyanobacterium Synechococcus sp., the green alga Tetraselmis suecica and three algae containing chlorophyll a+c (the diatom Thalassiosira weissflogii, the dinoflagellate Protoceratium reticulatum and the coccolithophorid Emiliania huxleyi) were grown in media containing 1, 5, 10, 20, or 30 mm SO(4) (2-) . The cyanobacterium and the green alga showed no growth response to varying [SO(4) (2-) ]. By contrast, the three chlorophyll a+c algae showed improved growth with higher [SO(4) (2-) ], but only up to 10 mm. The chlorophyll a+c algae, but not the green alga or cyanobacterium, also showed lower C:S with higher [SO(4) (2-) ]. When the same experiment was repeated in the presence of a ciliate predator (Euplotes sp.), T. suecica and T. weissflogii increased their specific growth rate in most treatments, whereas the growth rate of Synechococcus sp. was not affected or decreased in the presence of grazers. In a third experiment, T. suecica, T. weissflogii, P. reticulatum and Synechococcus sp. were grown in conditions approximating modern, earlier Paleozoic and Proterozoic seawater. In these treatments, sulfate availability, nitrogen source, metal availability and Pco(2) varied. Monospecific cultures exhibited their highest growth rates in the Proterozoic treatment. In mixed culture, T. weissflogii outgrew other species in modern seawater and T.suecica outgrew the others in Paleozoic water. Synechococcus sp. grew best in Proterozoic seawater, but did not outgrow eukaryotic species in any treatment. Collectively, our results suggest that secular increase in seawater [SO(4) (2-) ] may have facilitated the evolutionary expansion of chlorophyll a+c phytoplankton, but probably not to the exclusion of other biological and environmental factors. PMID- 21627762 TI - Human response to unintended intrathecal injection of botulinum toxin. AB - OBJECTIVE: Describe the first reported human intrathecal (IT) botulinum toxin injection. DESIGN: Case report. SETTING AND PATIENTS: We report here the sequelae to an unintended IT injection of botulinum toxin type B (BTB) in a 60-year-old woman with chronic back pain. RESULTS: Following the IT administration of BTB, the patient experienced the onset of symmetric ascending stocking distribution painful dysesthesias, which persisted for approximately 6 months before receding. Objective neurologic deficits were not appreciated, and analgesic effects were prominently absent. CONCLUSIONS: Analgesic actions of botulinum toxins in animals and in humans have led to speculation that IT botulinum toxin might exert significant analgesic effects. The unusual and unexpected subsequent clinical course, neurologic sequelae, dysesthesias, and absence of analgesia suggest that botulinum toxin will not be a therapeutic modality to treat pain as proposed by those studying botulinum toxin in animal models. PMID- 21627763 TI - Revisiting Paulozzi et al.'s "Prescription drug monitoring programs and death rates from drug overdose". PMID- 21627764 TI - Abnormal uterine bleeding: is it an under-reported side effect after epidural steroid injection for the management of low back pain? PMID- 21627765 TI - Subjective sleep quality and ethnicity are interactively related to standard and situation-specific measures of pain catastrophizing. AB - OBJECTIVE: Sleep quality and ethnicity are related to a host of general health outcomes including the experience of pain, yet it remains unclear whether poor sleep quality and ethnicity might interactively affect pain catastrophizing and laboratory-evoked acute pain reports. The current study examined the cross sectional associations of subjective sleep quality, ethnicity, and their interaction with pain catastrophizing and pain reports. DESIGN: Healthy (N = 149), ethnically diverse (58% Caucasian American, 23% Asian American, 19% African American) young adults were subjected to a cold pressor task (CPT). Prior to CPT, participants completed the Pittsburgh Sleep Quality Index and a standard version of the Pain Catastrophizing Scale (PCS). Following CPT, participants completed a situation-specific version of the PCS. RESULTS: Adjusted analyses revealed a significant sleep quality by ethnicity interaction for standard catastrophizing reports. Particularly, African Americans with poor overall sleep quality reported the greatest level of catastrophizing on the standard PCS relative to their Caucasian American and Asian American counterparts. Furthermore, African Americans with poorer sleep efficiency reported greater catastrophizing on the situation-specific PCS compared with Caucasian American and Asian Americans. Catastrophizing was significantly correlated with pain reports. CONCLUSIONS: These results suggest that African Americans with poorer sleep quality may be at greater risk for catastrophizing, a known contributor to more intense pain and increased pain-related emotional distress. Whether interventions that improve the sleep quality of ethnic minorities affect pain catastrophizing is in need of investigation. PMID- 21627766 TI - Efficacy of gabapentin enacarbil vs placebo in patients with postherpetic neuralgia and a pharmacokinetic comparison with oral gabapentin. AB - BACKGROUND: The efficacy of gabapentin in some patients with postherpetic neuralgia (PHN) may be limited by suboptimal drug exposure from unpredictable and saturable absorption. Gabapentin enacarbil (GEn) was designed for absorption by high-capacity transporters expressed throughout the intestine and undergoes rapid postabsorption hydrolysis to gabapentin. GEn extended-release tablets provide sustained, dose-proportional gabapentin exposure. This study assessed the efficacy of GEn vs placebo and compared the pharmacokinetics of gabapentin after oral dosing of GEn or gabapentin in patients with PHN. METHODS: In this double blind, randomized study, 115 patients with PHN completed a 7-day baseline period and 11-day gabapentin run-in period. Eligible patients were randomized and 101 received double-blind GEn 1,200 mg (624 mg-equivalents gabapentin) (n = 47) or placebo (n = 54), twice daily for 14 days. We evaluated patient-reported pain, sleep, mood, global improvement, and adverse events, plus gabapentin pharmacokinetics. RESULTS: The improvement in mean weekly pain scores from baseline to the end of treatment (primary endpoint) was significantly greater for GEn (-2.1) vs placebo (-1.2), P = 0.0321. Significant improvements from GEn vs placebo were also seen in sleep, mood, and patient global assessment (P < 0.05). With a 31% lower daily dose of gabapentin equivalents, GEn tablets provided a significant increase in average steady state gabapentin concentrations vs gabapentin capsules in the same patients (n = 42; P = 0.0050). CONCLUSIONS: GEn was effective in providing PHN pain relief, improved gabapentin exposure compared with gabapentin capsules, and was generally safe and well tolerated in patients with PHN. PMID- 21627767 TI - Microcurrent transcutaneous electric nerve stimulation in painful diabetic neuropathy: a randomized placebo-controlled study. AB - OBJECTIVES: Diabetes is a common health care problem in western countries. Painful diabetic neuropathy (PDN) might be one of the consequences of long ongoing diabetes; it is estimated that approximately 20% of European diabetic patients suffer from PDN. Transcutaneous electrical nerve stimulation (TENS) is often used as additional pain treatment. However, recent studies show inconsistent results. We aimed to assess the effect of micro-TENS in reducing neuropathic pain in patients with PDN in a placebo-controlled, single-blinded, and randomized design. DESIGN/SETTING/PATIENTS/OUTCOME MEASURES: 22 diabetic patients have been treated with a micro-TENS therapy and 19 patients have been treated with a placebo therapy. Treatment duration was 4 weeks with three therapeutical settings per week. Standardized questionnaires (Pain Disability Index [PDI], neuropathic pain score [NPS], Center for Epidemiologic Studies Depression Scale [CES-D]) were used to assess pain intensity, pain disability, as well as quality of life at baseline at the end of the treatment period and 4 weeks after treatment termination. RESULTS: Patients with a minimum of 30% reduction in NPS were defined as therapy responders. After 4 weeks of treatment, 6/21 patients in the verum group vs 10/19 patients in the placebo group responded to therapy. The median PDI score after 4 weeks of treatment showed a reduction of 23% in the verum vs 25% in the placebo group. The differences did not reach statistical significance. CONCLUSIONS: The pain reduction with the applied transcutaneous electrotherapy regimen is not superior to a placebo treatment. PMID- 21627768 TI - Perioperative intravenous acetaminophen and NSAIDs. AB - BACKGROUND: Unrelieved postoperative pain may result in pain/suffering, as well as multiple physiological and psychological consequences (e.g., splinting, impaired gastrointestinal motility/ileus, and impaired wound healing) which may adversely affect perioperative outcomes and contribute to increased length of stay. Multimodal or balanced analgesia, utilizing regional analgesic techniques (where possible) and nonopioid analgesics appear to represent a viable strategy to decrease systemic opioid consumption and improve postoperative analgesia. The use of multimodal analgesic strategies may result in reduced frequency and severity of unwanted opioid-related adverse effects, better clinically meaningful pain relief, diminished opioid consumption, and an overall improvement of patient satisfaction as well as health outcomes (e.g., earlier ambulation and discharge). OBJECTIVES: Review key aspects of intravenous (i.v.) acetaminophen (APAP) use in the postoperative setting. DESIGN: Focused literature review. RESULTS: Intravenous APAP is safe, effective for mild-to-moderate postoperative pain, well tolerated, and has a very favorable side effect profile with no clearly demonstrated clinically significant drug-drug interactions. It does not exhibit any significant effects on platelet aggregation and therefore may be the preferred nonopioid analgesic when surgical bleeding is an issue. CONCLUSION: The i.v. formulation of APAP represents a safe and effective first-line analgesic agent for the treatment of acute mild-to-moderate pain in the perioperative setting when oral agents may be impractical or when rapid onset with predictable therapeutic dosing is required. PMID- 21627769 TI - An efficient method to optimize Kluyveromyces lactis gene targeting. AB - Kluyveromyces lactis strains impaired in the nonhomologous end-joining pathway are relevant tools for the homologous integration of exogenous DNA into the genome, as in the mutant strains, close to 100% of the integrants are targeted to the homologous locus, compared with a few per cent for the wild-type recipient. Using a loxP-kanMX-loxP cassette together with a Cre-recombinase plasmid, a nej1?loxP mutant strain suitable for multiple gene disruption has been constructed. Furthermore, using this strain, PCR-generated constructs with only 50 bp of homologous flanking sequences resulted in efficient exogenous DNA targeting. PMID- 21627770 TI - [S3-guidelines for the treatment of psoriasis vulgaris Update 2011]. PMID- 21627772 TI - [S3-guidelines for the treatment of psoriasis vulgaris--methods report]. AB - BACKGROUND: This methods report summarizes the methodology used to update the German Psoriasis Guidelines. METHODS: The guidelines were developed following the recommendations of the Association of the Scientific Medical Societies in Germany. Medline, Cochrane Library and Embase were searched to generate new evidence. In addition, the results from the literature search from the EU Psoriasis Guidelines were used. The recommendations were discussed during a consensus conference using nominal group technique and were voted on during the Delphi procedure. An extensive internal and external review (open) was performed. RESULTS: Due to changes in drug licensing efalizumab was excluded from the guidelines and adalimumab and ustekinumab were added. 97 new studies were included to serve as a basis for the recommendations. The level of evidence improved for calcineurin inhibitors from level 4 to level 2/3 and for MTX and systemic retinoids from level 3 to level 2. A lack of evidence still exists for coal tar (level of evidence 4). All other described interventions have a level of evidence of 2 or 1. CONCLUSIONS: The field of psoriasis therapy is in constant transition. A lack of head to head trials makes direct comparisons still a challenging task. Continuous updating will be necessary to respond to the further changes expected in the field of psoriasis. PMID- 21627773 TI - Systemic treatment with corticosteroids in psoriasis--health care provision far beyond the S3-guidelines. AB - BACKGROUND: The basis of induction therapy for psoriasis in Germany is the S3 guideline which for moderate to severe psoriasis recommends the systemic agents fumaric acid esters, methotrexate, cyclosporine and the biologicals infliximab, etanercept, adalimumab and ustekinumab. Systemic glucocorticosteroids (GCS) are not proposed. QUESTION: To what extend are systemic glucocorticosteroids prescribed in psoriasis therapy? METHODS: The database of a German nationwide statutory health insurance 2007 was analyzed. Prescriptions of systemic glucocorticosteroids were identified by ATC-encoding. Psoriasis patients with comorbidities requiring systemic glucocorticosteroid medication were analyzed separately. RESULTS: Among 1,423,308 continuously insured patients in 2007 n = 34,728 (2.4 %) were patients with psoriasis. The most prescribed systemic agents were corticosteroids (2774 patients), followed by methotrexate (853), fumaric acid esters (342), retinoids (110) and cyclosporine A (105). Even after excluding patients with psoriatic arthritis, systemic corticosteroids were still the most often prescribed systemic drugs. The average number of DDD (Defined Daily Doses) per insured patient was 120 (general practitioners), 141 (internists) and 68 (dermatologists). These results were confirmed by a control analysis with IMS data. CONCLUSIONS: In Germany, psoriasis is often treated with systemic corticosteroids though these are not recommended by the S3-guideline. General practitioners and internists are the main prescribers. PMID- 21627774 TI - Cetuximab-associated folliculitis predominantly affecting the lower limbs. PMID- 21627775 TI - Permanent genetic resources added to Molecular Ecology Resources Database 1 February 2011-31 March 2011. AB - This article documents the addition of 111 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Acipenser oxyrinchus desotoi, Anopheles nuneztovari sensu lato, Asellus aquaticus, Calopteryx splendens, Calopteryx virgo, Centaurea aspera, Centaurea seridis, Chilina dombeyana, Proctoeces cf. lintoni and Pyrenophora teres f. teres. PMID- 21627777 TI - The German Young Olympic Athletes' Lifestyle and Health Management Study (GOAL Study): design of a mixed-method study. AB - BACKGROUND: In order to perform at top levels, elite athletes have to both protect and risk their health at the same time. Adolescent elite athletes have the additional challenge of coping with substantial physical, psychological and social transformations. The contradictory phenomenon of protecting and risking the adolescent athletes' health in sports challenges the development of health promotion and protection strategies. The GOAL Study (German Young Olympic Athletes' Lifestyle and Health Management Study) analyzes the individual and organizational management of health in adolescent elite sports. METHODS/DESIGN: We combine quantitative and qualitative approaches in a mixed-method study. This allows us to gather a broad range of representative information on squad athletes from all Olympic disciplines as well as in-depth information on four selected Olympic disciplines (artistic gymnastics, biathlon, handball and wrestling). Within the quantitative section we attempt to identify the young athletes' health and nutrition behavior, their subjective health state and their lay health representations, health-related social networks, and structures of medical attendance. 1138 national team level athletes born between 1992 and 1995 from 51 Olympic disciplines responded to the questionnaire (response rate: 61,75%). The qualitative section investigates the meaning and relevance of health and nutrition within the athletes' sports specific surroundings, the impact of biographic backgrounds on individual health behavior, and sports specific cultures of health, nutrition and risk. We interviewed 24 athletes and 28 coaching and medical experts, and carried out 14 multi-day participant observations at training sessions and competitions. CONCLUSIONS: The studies' results will serve as the basis for developing tailored health promotion strategies to be in cooperation with German elite sports associations. PMID- 21627776 TI - Mobile DNA and the TE-Thrust hypothesis: supporting evidence from the primates. AB - Transposable elements (TEs) are increasingly being recognized as powerful facilitators of evolution. We propose the TE-Thrust hypothesis to encompass TE facilitated processes by which genomes self-engineer coding, regulatory, karyotypic or other genetic changes. Although TEs are occasionally harmful to some individuals, genomic dynamism caused by TEs can be very beneficial to lineages. This can result in differential survival and differential fecundity of lineages. Lineages with an abundant and suitable repertoire of TEs have enhanced evolutionary potential and, if all else is equal, tend to be fecund, resulting in species-rich adaptive radiations, and/or they tend to undergo major evolutionary transitions. Many other mechanisms of genomic change are also important in evolution, and whether the evolutionary potential of TE-Thrust is realized is heavily dependent on environmental and ecological factors. The large contribution of TEs to evolutionary innovation is particularly well documented in the primate lineage. In this paper, we review numerous cases of beneficial TE-caused modifications to the genomes of higher primates, which strongly support our TE Thrust hypothesis. PMID- 21627778 TI - The occupational risk of Helicobacter pylori infection among gastroenterologists and their assistants. AB - BACKGROUND: Helicobacter pylori is a widely spread bacterium that mainly inhabits the gastric mucosa and can lead to serious illnesses such as peptic ulcer disease, gastric carcinoma and gastric MALT lymphoma. The oral-oral route seems to be the main transmission route. The fact that endoscopes are contaminated after being used to perform a gastroscopy leads one to question whether gastroenterologists and endoscopy nurses and assistants run a higher risk of infection. METHODS: A systematic search for literature was conducted in the MEDLINE and EMBASE databases and further publications were found in reference lists of relevant articles. Epidemiological studies on the occupational exposure of endoscopy personnel were collected and their quality was assessed. Pooled effect estimates were identified in a meta-analysis. RESULTS: Of the 24 studies included in the analysis, 15 were considered to be methodologically good. Of these 15 studies, eight single studies showed a statistically significant increased risk of infection for gastroenterologists, and five for their assistants. Meta-analysis across all methodologically good studies found a statistically significant risk of 1.6 (95%CI 1.3-2.0) for doctors. The pooled effect estimates also indicated a statistically significant risk of Helicobacter pylori infection (RR 1.4; 95%CI 1.1-1.8) for assistants too.When studies are stratified by medical and non-medical control groups, statistically significant risks can only be recognised in the comparison with non-medical controls. CONCLUSIONS: In summary, our results demonstrated an increased risk of Helicobacter pylori infection among gastroenterological personnel. However, the choice of control group is important for making a valid assessment of occupational exposure risks. PMID- 21627780 TI - Dietary differences between elderly Iranians living in Sweden and Iran a cross sectional comparative study. AB - BACKGROUND: During the last decades, global migration has increased and many immigrant groups have a higher prevalence than the native born population of several cardiovascular disease risk factors, including poor dietary habits. However, it is uncertain if dietary habits in immigrant populations reflect dietary habits in their country of origin or if the current diet is a consequence of the migration and possible change of dietary habits. The aim of this study was to examine possible dietary differences between elderly Iranians living in Stockholm, Sweden with elderly Iranians living in Tehran, Iran, taking into account sex, age, marital status, and education. METHODS: Dietary intakes were assessed by semi--quantitative food frequency questionnaire in a cross-sectional study of 121 Iranians living in Stockholm and 52 Iranians living in Tehran, aged 60-80. Differences in dietary habits between the two groups was analysed by bootstrapped regression analyses with 1000 replications. RESULTS: Iranians living in Sweden had significantly higher intake of protein, total fat, fiber than Iranians living in Iran, but lower consumption of carbohydrates. The observed differences in intake of macronutrients were reflected in consumed amount of all food items, which were higher among Iranians living in Iran with the exception of bread and grain consumption which was lower. CONCLUSIONS: There are general differences in dietary habits between Iranians living in Iran and Iranians living in Sweden. Parts of observed differences in dietary habits may reflect a favourable adoption process to the Swedish dietary habits after migration. Meanwhile other differences are point of concern in light of the high prevalence of overweight, among Iranians living in Sweden and can have unfavourable impact in particular in the context of cardiovascular health. PMID- 21627779 TI - The membrane-spanning 4-domains, subfamily A (MS4A) gene cluster contains a common variant associated with Alzheimer's disease. AB - BACKGROUND: In order to identify novel loci associated with Alzheimer's disease (AD), we conducted a genome-wide association study (GWAS) in the Spanish population. METHODS: We genotyped 1,128 individuals using the Affymetrix Nsp I 250K chip. A sample of 327 sporadic AD patients and 801 controls with unknown cognitive status from the Spanish general population were included in our initial study. To increase the power of the study, we combined our results with those of four other public GWAS datasets by applying identical quality control filters and the same imputation methods, which were then analyzed with a global meta-GWAS. A replication sample with 2,200 sporadic AD patients and 2,301 controls was genotyped to confirm our GWAS findings. RESULTS: Meta-analysis of our data and independent replication datasets allowed us to confirm a novel genome-wide significant association of AD with the membrane-spanning 4-domains subfamily A (MS4A) gene cluster (rs1562990, P = 4.40E-11, odds ratio = 0.88, 95% confidence interval 0.85 to 0.91, n = 10,181 cases and 14,341 controls). CONCLUSIONS: Our results underscore the importance of international efforts combining GWAS datasets to isolate genetic loci for complex diseases. PMID- 21627781 TI - Recommendations to improve physical activity among teenagers--a qualitative study with ethnic minority and European teenagers. AB - BACKGROUND: To understand the key challenges and explore recommendations from teenagers to promote physical activity with a focus on ethnic minority children. METHODS: Focus groups with teenagers aged 16-18 of Bangladeshi, Somali or Welsh descent attending a participating school in South Wales, UK. There were seventy four participants (18 Somali, 24 Bangladeshi and 32 Welsh children) divided into 12 focus groups. RESULTS: The boys were more positive about the benefits of exercise than the girls and felt there were not enough facilities or enough opportunity for unsupervised activity. The girls felt there was a lack of support to exercise from their family. All the children felt that attitudes to activity for teenagers needed to change, so that there was more family and community support for girls to be active and for boys to have freedom to do activities they wanted without formal supervision. It was felt that older children from all ethnic backgrounds should be involved more in delivering activities and schools needs to provide more frequent and a wider range of activities. CONCLUSIONS: This study takes a child-focused approach to explore how interventions should be designed to promote physical activity in youth. Interventions need to improve access to facilities but also counteract attitudes that teenagers should be studying or working and not 'hanging about' playing with friends. Thus, the value of activity for teenagers needs to be promoted not just among the teenagers but with their teachers, parents and members of the community. PMID- 21627782 TI - The mitochondrial genome sequence of the ciliate Paramecium caudatum reveals a shift in nucleotide composition and codon usage within the genus Paramecium. AB - BACKGROUND: Despite the fact that the organization of the ciliate mitochondrial genome is exceptional, only few ciliate mitochondrial genomes have been sequenced until today. All ciliate mitochondrial genomes are linear. They are 40 kb to 47 kb long and contain some 50 tightly packed genes without introns. Earlier studies documented that the mitochondrial guanine + cytosine contents are very different between Paramecium tetraurelia and all studied Tetrahymena species. This raises the question of whether the high mitochondrial G+C content observed in P. tetraurelia is a characteristic property of Paramecium mtDNA, or whether it is an exception of the ciliate mitochondrial genomes known so far. To test this question, we determined the mitochondrial genome sequence of Paramecium caudatum and compared the gene content and sequence properties to the closely related P. tetraurelia. RESULTS: The guanine + cytosine content of the P. caudatum mitochondrial genome was significantly lower than that of P. tetraurelia (22.4% vs. 41.2%). This difference in the mitochondrial nucleotide composition was accompanied by significantly different codon usage patterns in both species, i.e. within P. caudatum clearly A/T ending codons dominated, whereas for P. tetraurelia the synonymous codons were more balanced with a higher number of G/C ending codons. Further analyses indicated that the nucleotide composition of most members of the genus Paramecium resembles that of P. caudatum and that the shift observed in P. tetraurelia is restricted to the P. aurelia species complex. CONCLUSIONS: Surprisingly, the codon usage bias in the P. caudatum mitochondrial genome, exemplified by the effective number of codons, is more similar to the distantly related T. pyriformis and other single-celled eukaryotes such as Chlamydomonas, than to the closely related P. tetraurelia. These differences in base composition and codon usage bias were, however, not reflected in the amino acid composition. Most probably, the observed picture is best explained by a hitherto unknown (neutral or adaptive) mechanism that increased the guanine + cytosine content in P. tetraurelia mtDNA on the one hand, and strong purifying selection on the ancestral amino acid composition on the other hand. These contradicting forces are counterbalanced by a considerably altered codon usage pattern. PMID- 21627783 TI - Human nucleosomes: special role of CG dinucleotides and Alu-nucleosomes. AB - BACKGROUND: The periodical occurrence of dinucleotides with a period of 10.4 bases now is undeniably a hallmark of nucleosome positioning. Whereas many eukaryotic genomes contain visible and even strong signals for periodic distribution of dinucleotides, the human genome is rather featureless in this respect. The exact sequence features in the human genome that govern the nucleosome positioning remain largely unknown. RESULTS: When analyzing the human genome sequence with the positional autocorrelation method, we found that only the dinucleotide CG shows the 10.4 base periodicity, which is indicative of the presence of nucleosomes. There is a high occurrence of CG dinucleotides that are either 31 (10.4 * 3) or 62 (10.4 * 6) base pairs apart from one another - a sequence bias known to be characteristic of Alu-sequences. In a similar analysis with repetitive sequences removed, peaks of repeating CG motifs can be seen at positions 10, 21 and 31, the nearest integers of multiples of 10.4. CONCLUSIONS: Although the CG dinucleotides are dominant, other elements of the standard nucleosome positioning pattern are present in the human genome as well.The positional autocorrelation analysis of the human genome demonstrates that the CG dinucleotide is, indeed, one visible element of the human nucleosome positioning pattern, which appears both in Alu sequences and in sequences without repeats. The dominant role that CG dinucleotides play in organizing human chromatin is to indicate the involvement of human nucleosomes in tuning the regulation of gene expression and chromatin structure, which is very likely due to cytosine methylation/-demethylation in CG dinucleotides contained in the human nucleosomes. This is further confirmed by the positions of CG-periodical nucleosomes on Alu sequences. Alu repeats appear as monomers, dimers and trimers, harboring two to six nucleosomes in a run. Considering the exceptional role CG dinucleotides play in the nucleosome positioning, we hypothesize that Alu nucleosomes, especially, those that form tightly positioned runs, could serve as "anchors" in organizing the chromatin in human cells. PMID- 21627784 TI - Bone regeneration: current concepts and future directions. AB - Bone regeneration is a complex, well-orchestrated physiological process of bone formation, which can be seen during normal fracture healing, and is involved in continuous remodelling throughout adult life. However, there are complex clinical conditions in which bone regeneration is required in large quantity, such as for skeletal reconstruction of large bone defects created by trauma, infection, tumour resection and skeletal abnormalities, or cases in which the regenerative process is compromised, including avascular necrosis, atrophic non-unions and osteoporosis. Currently, there is a plethora of different strategies to augment the impaired or 'insufficient' bone-regeneration process, including the 'gold standard' autologous bone graft, free fibula vascularised graft, allograft implantation, and use of growth factors, osteoconductive scaffolds, osteoprogenitor cells and distraction osteogenesis. Improved 'local' strategies in terms of tissue engineering and gene therapy, or even 'systemic' enhancement of bone repair, are under intense investigation, in an effort to overcome the limitations of the current methods, to produce bone-graft substitutes with biomechanical properties that are as identical to normal bone as possible, to accelerate the overall regeneration process, or even to address systemic conditions, such as skeletal disorders and osteoporosis. PMID- 21627785 TI - Novel variants in the PRDX6 Gene and the risk of Acute Lung Injury following major trauma. AB - BACKGROUND: Peroxiredoxin 6 (PRDX6) is involved in redox regulation of the cell and is thought to be protective against oxidant injury. Little is known about genetic variation within the PRDX6 gene and its association with acute lung injury (ALI). In this study we sequenced the PRDX6 gene to uncover common variants, and tested association with ALI following major trauma. METHODS: To examine the extent of variation in the PRDX6 gene, we performed direct sequencing of the 5' UTR, exons, introns and the 3' UTR in 25 African American cases and controls and 23 European American cases and controls (selected from a cohort study of major trauma), which uncovered 80 SNPs. In silico modeling was performed using Patrocles and Transcriptional Element Search System (TESS). Thirty seven novel and tagging SNPs were tested for association with ALI compared with ICU at risk controls who did not develop ALI in a cohort study of 259 African American and 254 European American subjects that had been admitted to the ICU with major trauma. RESULTS: Resequencing of critically ill subjects demonstrated 43 novel SNPs not previously reported. Coding regions demonstrated no detectable variation, indicating conservation of the protein. Block haplotype analyses reveal that recombination rates within the gene seem low in both Caucasians and African Americans. Several novel SNPs appeared to have the potential for functional consequence using in silico modeling. Chi2 analysis of ALI incidence and genotype showed no significant association between the SNPs in this study and ALI. Haplotype analysis did not reveal any association beyond single SNP analyses. CONCLUSIONS: This study revealed novel SNPs within the PRDX6 gene and its 5' and 3' flanking regions via direct sequencing. There was no association found between these SNPs and ALI, possibly due to a low sample size, which was limited to detection of relative risks of 1.93 and above. Future studies may focus on the role of PRDX6 genetic variation in other diseases, where oxidative stress is suspected. PMID- 21627786 TI - Neuromuscular abnormality and autonomic dysfunction in patients with cerebrotendinous xanthomatosis. AB - BACKGROUND: Cerebrotendinous xanthomatosis (CTX) is a rare lipid-storage disease. Neuromuscular abnormality and autonomic system (ANS) dysfuction in CTX are rarely examined in large-scale studies in the literature. We studied the peripheral nervous system, myopathology, and autonomic system of four CTX patients and performed a literature review of the reported CTX patients with peripheral neuropathy. METHODS: Four biochemically and genetically confirmed CTX patients, belonging to two families, were included for study and all received nerve conduction study (NCS), muscle biopsy for histopathologic and ultrastructural study, skin biopsy for intraepidermal nerve fiber (INEF) density measurement, autonomic testings including sympathetic skin response, R-R interval variation and head-up tilt test using an automated tilt table to record the changes of blood pressure and heart rate in different postures. The Q-Sweat test was also applied for the detection of sweat amount and onset time of response. The clinical characteristics, study methods and results of 13 studies of peripheral neuropathy in CTX patients in the literature were also recorded for analysis. RESULTS: The results of NCS study showed axonal sensory-motor polyneuropathy in three CTX cases and mixed axonal and demyelinating sensor-motor polyneuropathy in one. The myopathological and histopathologic studies revealed mild denervation characteristics, but the ultrastructural study revealed changes of mitochondria and the membranous system, and increased amounts of glycogen, lipofuscin and lipid deposition. The ANS study revealed different degrees of abnormalities in the applied tests and the INEF density measurement showed small fiber neuropathy in three of the four CTX patients. The literature review of peripheral neuropathy in CTX revealed different types of peripheral neuropathy, of which axonal peripheral neuropathy was the most common. CONCLUSIONS: Peripheral neuropathy, especially the subtype of axonal sensori-motor neuropathy, is common in patients with CTX. Evidence of lipid metabolic derangement in CTX can be reflected in ultrastructural studies of muscles. With an adequate multi-parametric evaluation, a high incidence of ANS abnormalities can be seen in this rare lipid-storage disease, and a high incidence of small fiber involvement is also reflected in the IENF density measurement of skin biopsies. PMID- 21627787 TI - FUMEPOC: early detection of chronic obstructive pulmonary disease in smokers. AB - BACKGROUND: Currently is not feasible using conventional spirometry as a screening method in Primary Care especially among smoking population to detect chronic obstructive pulmonary disease in early stages. Therefore, the FUMEPOC study protocol intends to analyze the validity and reliability of Vitalograph COPD-6 spirometer as simpler tool to aid screening and diagnosis of this disease in early stages in primary care surgery. METHODS / DESIGN: STUDY DESIGN: An observational, descriptive study of diagnostic tests, undertaken in Primary Care and Pneumology Outpatient Care Centre at San Juan Hospital and Elda Hospital. All smokers attending the primary care surgery and consent to participate in the study will undergo a test with Vitalograph COPD-6 spirometer. Subsequently, a conventional spirometry will be performed in the hospital and the results will be compared with those of the Vitalograph COPD-6 test. DISCUSSION: It is difficult to use the spirometry as screening for early diagnose test in real conditions of primary care clinical practice. The use of a simpler tool, Vitalograph COPD-6 spirometer, can help in the early diagnose and therefore, it could improve the clinical management of the disease. PMID- 21627788 TI - Impact of bleeding-related complications and/or blood product transfusions on hospital costs in inpatient surgical patients. AB - BACKGROUND: Inadequate surgical hemostasis may lead to transfusion and/or other bleeding-related complications. This study examines the incidence and costs of bleeding-related complications and/or blood product transfusions occurring as a consequence of surgery in various inpatient surgical cohorts. METHODS: A retrospective analysis was conducted using Premier's PerspectiveTM hospital database. Patients who had an inpatient procedure within a specialty of interest (cardiac, vascular, non-cardiac thoracic, solid organ, general, reproductive organ, knee/hip replacement, or spinal surgery) during 2006-2007 were identified. For each specialty, the rate of bleeding-related complications (including bleeding event, intervention to control for bleeding, and blood product transfusions) was examined, and hospital costs and length of stay (LOS) were compared between surgeries with and without bleeding-related complications. Incremental costs and ratios of average total hospital costs for patients with bleeding-related complications vs. those without complications were estimated using ordinary least squares (OLS) regression, adjusting for demographics, hospital characteristics, and other baseline characteristics. Models using generalized estimating equations (GEE) were also used to measure the impact of bleeding-related complications on costs while accounting for the effects related to the clustering of patients receiving care from the same hospitals. RESULTS: A total of 103,829 cardiac, 216,199 vascular, 142,562 non-cardiac thoracic, 45,687 solid organ, 362,512 general, 384,132 reproductive organ, 246,815 knee/hip replacement, and 107,187 spinal surgeries were identified. Overall, the rate of bleeding-related complications was 29.9% and ranged from 7.5% to 47.4% for reproductive organ and cardiac, respectively. Overall, incremental LOS associated with bleeding-related complications or transfusions (unadjusted for covariates) was 6.0 days and ranged from 1.3 to 9.6 days for knee/hip replacement and non cardiac thoracic, respectively. The incremental cost per hospitalization associated with bleeding-related complications and adjusted for covariates was highest for spinal surgery ($17,279) followed by vascular ($15,123), solid organ ($13,210), non-cardiac thoracic ($13,473), cardiac ($10,279), general ($4,354), knee/hip replacement ($3,005), and reproductive organ ($2,805). CONCLUSIONS: This study characterizes the increased hospital LOS and cost associated with bleeding related complications and/or transfusions occurring as a consequence of surgery, and supports implementation of blood-conservation strategies. PMID- 21627789 TI - RAG: an update to the RNA-As-Graphs resource. AB - BACKGROUND: In 2004, we presented a web resource for stimulating the search for novel RNAs, RNA-As-Graphs (RAG), which classified, catalogued, and predicted RNA secondary structure motifs using clustering and build-up approaches. With the increased availability of secondary structures in recent years, we update the RAG resource and provide various improvements for analyzing RNA structures. DESCRIPTION: Our RAG update includes a new supervised clustering algorithm that can suggest RNA motifs that may be "RNA-like". We use this utility to describe RNA motifs as three classes: existing, RNA-like, and non-RNA-like. This produces 126 tree and 16,658 dual graphs as candidate RNA-like topologies using the supervised clustering algorithm with existing RNAs serving as the training data. A comparison of this clustering approach to an earlier method shows considerable improvements. Additional RAG features include greatly expanded search capabilities, an interface to better utilize the benefits of relational database, and improvements to several of the utilities such as directed/labeled graphs and a subgraph search program. CONCLUSIONS: The RAG updates presented here augment the database's intended function - stimulating the search for novel RNA functionality - by classifying available motifs, suggesting new motifs for design, and allowing for more specific searches for specific topologies. The updated RAG web resource offers users a graph-based tool for exploring available RNA motifs and suggesting new RNAs for design. PMID- 21627790 TI - Mutation of a nicotinic acetylcholine receptor beta subunit is associated with resistance to neonicotinoid insecticides in the aphid Myzus persicae. AB - BACKGROUND: Myzus persicae is a globally important aphid pest with a history of developing resistance to insecticides. Unusually, neonicotinoids have remained highly effective as control agents despite nearly two decades of steadily increasing use. In this study, a clone of M. persicae collected from southern France was found, for the first time, to exhibit sufficiently strong resistance to result in loss of the field effectiveness of neonicotinoids. RESULTS: Bioassays, metabolism and gene expression studies implied the presence of two resistance mechanisms in the resistant clone, one based on enhanced detoxification by cytochrome P450 monooxygenases, and another unaffected by a synergist that inhibits detoxifying enzymes. Binding of radiolabeled imidacloprid (a neonicotinoid) to whole body membrane preparations showed that the high affinity [3H]-imidacloprid binding site present in susceptible M. persicae is lost in the resistant clone and the remaining lower affinity site is altered compared to susceptible clones. This confers a significant overall reduction in binding affinity to the neonicotinoid target: the nicotinic acetylcholine receptor (nAChR). Comparison of the nucleotide sequence of six nAChR subunit (Mpalpha1-5 and Mpbeta1) genes from resistant and susceptible aphid clones revealed a single point mutation in the loop D region of the nAChR beta1 subunit of the resistant clone, causing an arginine to threonine substitution (R81T). CONCLUSION: Previous studies have shown that the amino acid at this position within loop D is a key determinant of neonicotinoid binding to nAChRs and this amino acid change confers a vertebrate-like character to the insect nAChR receptor and results in reduced sensitivity to neonicotinoids. The discovery of the mutation at this position and its association with the reduced affinity of the nAChR for imidacloprid is the first example of field-evolved target-site resistance to neonicotinoid insecticides and also provides further validation of exisiting models of neonicotinoid binding and selectivity for insect nAChRs. PMID- 21627791 TI - Hospital and outpatient clinic utilization among older people in the 3-5 years following the initiation of continuing care: a longitudinal cohort study. AB - BACKGROUND: Few studies have investigated the subsequent rate of hospital and outpatient clinic utilization in those who receive continuing care and have documented frequent usage over one year. Such knowledge may be helpful in identifying those who would benefit from preventive interventions. The aim of this study was to investigate and compare the subsequent rate of hospital and outpatient clinic utilization among older people with 0, 1, 2, 3 or more hospital stays in the first year following the initiation of continuing care. A further aim was to compare these groups regarding demographic data, health complaints, functional and cognitive ability, informal care and mortality. METHODS: A total of 1079 people, aged 65 years or older, who received a decision regarding the initiation of continuing care during the years 2001, 2002 or 2003 were investigated. Four groups were created based on whether they had 0, 1, 2 or >= 3 hospital stays in the first year following the initiation of continuing care and were investigated regarding the rate of hospital and outpatient clinic utilization in the subsequent 3-5 years. RESULTS: Fifty seven percent of the sample had no hospital stay during the first year following the initiation of continuing care, 20% had 1 stay, 10% had 2 stays and 13% had three or more hospital stays (range: 3-13). Those with >= 3 hospital stays in the first year continued to have the significantly highest rate of hospital and outpatient care utilization in the subsequent years. This group accounted for 57% of hospital stays in the first year, 27% in the second year and 18% in the third year. In this group the risk of having >= 3 hospital stays in the second year was 27% and 12% in the third year. CONCLUSIONS: There is a clear need for interventions targeted on prevention of frequent hospital and outpatient clinic utilization among those who are high users of hospital care in the first year after the initiation of continuing care. Perhaps an increased availability of medically skilled staff in the day to day care of these people in the municipalities could prevent frequent hospital and outpatient clinic utilization, especially hospital readmissions. PMID- 21627792 TI - Complete Columbian mammoth mitogenome suggests interbreeding with woolly mammoths. AB - BACKGROUND: Late Pleistocene North America hosted at least two divergent and ecologically distinct species of mammoth: the periglacial woolly mammoth (Mammuthus primigenius) and the subglacial Columbian mammoth (Mammuthus columbi). To date, mammoth genetic research has been entirely restricted to woolly mammoths, rendering their genetic evolution difficult to contextualize within broader Pleistocene paleoecology and biogeography. Here, we take an interspecific approach to clarifying mammoth phylogeny by targeting Columbian mammoth remains for mitogenomic sequencing. RESULTS: We sequenced the first complete mitochondrial genome of a classic Columbian mammoth, as well as the first complete mitochondrial genome of a North American woolly mammoth. Somewhat contrary to conventional paleontological models, which posit that the two species were highly divergent, the M. columbi mitogenome we obtained falls securely within a subclade of endemic North American M. primigenius. CONCLUSIONS: Though limited, our data suggest that the two species interbred at some point in their evolutionary histories. One potential explanation is that woolly mammoth haplotypes entered Columbian mammoth populations via introgression at subglacial ecotones, a scenario with compelling parallels in extant elephants and consistent with certain regional paleontological observations. This highlights the need for multi-genomic data to sufficiently characterize mammoth evolutionary history. Our results demonstrate that the use of next-generation sequencing technologies holds promise in obtaining such data, even from non-cave, non-permafrost Pleistocene depositional contexts. PMID- 21627794 TI - Perceptions of young Jordanian adults to proposed anti-tobacco pictorial warning labels. AB - BACKGROUND: In commitment to the Framework Convention on Tobacco Control (FCTC), four new pictorial warnings are now being proposed for display on cigarette packages sold in Jordan. The aim of this study was to gauge the immediate perceptions of young Jordanian adults towards these new pictorials and compare these perceptions to those of the pictorial currently being used in the country. METHODS: A cross-sectional survey was conducted on a convenience sample of youth aged 17-26. The interviewer-administered survey gauged participants' perceptions of salience, fear elicitation, and gained information as well as participants' motivation to remain non-smokers or quit smoking after viewing each of the four proposed new pictorials as well as the current pictorial used in Jordan. Perceptions regarding each new pictorial were compared to the current pictorial. RESULTS: A total of 450 surveys were included in the analysis. The sample (mean age 20.9) was 51.6% female and 31.3% cigarette (regular or occasional) smokers. In smokers, only one proposed pictorial had significantly more smokers perceiving it as salient or adding to information when compared to the current pictorial. More smokers reported fear when observing the proposed pictorials compared with current pictorial, but overall proportions reporting fear were generally less than 50%. Furthermore, all new pictorials motivated significantly more smokers to consider quitting compared with the current pictorial; however, the overall proportion of smokers reporting motivation was < 25%. Among nonsmokers, significantly more respondents perceived the new pictorials as salient and fear eliciting compared to the old pictorial, but there were no major differences in information added. Motivation to remain non-smokers was comparable between the old and new pictorials. CONCLUSION: Given the variability of response across both smokers and nonsmokers, and across the three elements of perception (salience, added information, fear) for each pictorial, further testing of the pictorials in a more diverse sample of Jordanian young adults prior to launch is recommended. PMID- 21627793 TI - Quantifying differential gene connectivity between disease states for objective identification of disease-relevant genes. AB - BACKGROUND: Network modeling of whole transcriptome expression data enables characterization of complex epistatic (gene-gene) interactions that underlie cellular functions. Though numerous methods have been proposed and successfully implemented to develop these networks, there are no formal methods for comparing differences in network connectivity patterns as a function of phenotypic trait. RESULTS: Here we describe a novel approach for quantifying the differences in gene-gene connectivity patterns across disease states based on Graphical Gaussian Models (GGMs). We compare the posterior probabilities of connectivity for each gene pair across two disease states, expressed as a posterior odds-ratio (postOR) for each pair, which can be used to identify network components most relevant to disease status. The method can also be generalized to model differential gene connectivity patterns within previously defined gene sets, gene networks and pathways. We demonstrate that the GGM method reliably detects differences in network connectivity patterns in datasets of varying sample size. Applying this method to two independent breast cancer expression data sets, we identified numerous reproducible differences in network connectivity across histological grades of breast cancer, including several published gene sets and pathways. Most notably, our model identified two gene hubs (MMP12 and CXCL13) that each exhibited differential connectivity to more than 30 transcripts in both datasets. Both genes have been previously implicated in breast cancer pathobiology, but themselves are not differentially expressed by histologic grade in either dataset, and would thus have not been identified using traditional differential gene expression testing approaches. In addition, 16 curated gene sets demonstrated significant differential connectivity in both data sets, including the matrix metalloproteinases, PPAR alpha sequence targets, and the PUFA synthesis pathway. CONCLUSIONS: Our results suggest that GGM can be used to formally evaluate differences in global interactome connectivity across disease states, and can serve as a powerful tool for exploring the molecular events that contribute to disease at a systems level. PMID- 21627795 TI - Antibiotic consumption in children prior to diagnosis of asthma. AB - BACKGROUND: Asthma is difficult to diagnose in children and at times misdiagnosis of an infection can occur. However, little is known about the magnitude and patterns of antibiotic consumption in children with asthma relative to those without asthma. METHODS: Using population-based data, 128,872 children were identified with at least 6 years of follow-up. The adjusted rate-ratio (RR) of antibiotics dispensed to asthmatic as compared to non-asthmatic children was determined. RESULTS: At age six, the RR of antibiotic consumption for asthmatics compared to non-asthmatics varied between, 1.66 to 2.32, depending on the year of asthma diagnosis. Of the 18,864 children with asthma at ages 2-8, 52% (n = 9,841) had antibiotics dispensed in the 6 months prior to their index date of asthma diagnosis. The RR of antibiotic consumption in the 1 month prior to asthma diagnosis compared to 5 months prior was 1.66 (95% CI 1.60-1.71). The RR was lower in males compared to females (1.58 vs 1.77), and lower in those who received antibiotics in the first year of life relative to those that did not (1.60 vs. 1.76). CONCLUSIONS: There is higher antibiotic consumption in children with asthma compared to those without asthma. The pattern of antibiotic use suggests that diagnosis guidelines are difficult to follow in young children leading to misdiagnosis and over treatment with antibiotics. PMID- 21627796 TI - An unusual presentation of multiple cavitated lung metastases from colon carcinoma. AB - BACKGROUND: Consolidation with or without ground-glass opacity is the typical radiologic finding of lung metastases of adenocarcinoma from the gastrointestinal tract. Lung excavated metastases from gastrointestinal carcinoma are very rare. CASE PRESENTATION: The authors describe an unusual presentation of multiple cavitated lung metastases from colon adenocarcinoma and discuss the outcome of a patient. The absence both of symptoms and other disease localizations, the investigations related to different diagnostic hypotheses and the empirical treatments caused a delay in correct diagnosis. Only a transparietal biopsy revealed the neoplastic origin of nodules. CONCLUSIONS: This report demonstrates that although lung excavated metastases are described in literature, initial failure to reach a diagnosis is common. We would like to alert clinicians and radiologists to the possibility of unusual atypical features of pulmonary metastases from colon adenocarcinoma. PMID- 21627797 TI - Effect of local anaesthesia and/or analgesia on pain responses induced by piglet castration. AB - BACKGROUND: Surgical castration in male piglets is painful and methods that reduce this pain are requested. This study evaluated the effect of local anaesthesia and analgesia on vocal, physiological and behavioural responses during and after castration. A second purpose was to evaluate if herdsmen can effectively administer anaesthesia. METHODS: Four male piglets in each of 141 litters in five herds were randomly assigned to one of four treatments: castration without local anaesthesia or analgesia (C, controls), analgesia (M, meloxicam), local anaesthesia (L, lidocaine), or both local anaesthesia and analgesia (LM). Lidocaine (L, LM) was injected at least three minutes before castration and meloxicam (M, LM) was injected after castration. During castration, vocalisation was measured and resistance movements judged. Behaviour observations were carried out on the castration day and the following day. The day after castration, castration wounds were ranked, ear and skin temperature was measured, and blood samples were collected for analysis of acute phase protein Serum Amyloid A concentration (SAA). Piglets were weighed on the castration day and at three weeks of age. Sickness treatments and mortality were recorded until three weeks of age. RESULTS: Piglets castrated with lidocaine produced calls with lower intensity (p < 0.001) and less resistance movements (p < 0.001) during castration. Piglets that were given meloxicam displayed less pain-related behaviour (huddled up, spasms, rump-scratching, stiffness and prostrated) on both the castration day (p = 0.06, n.s.) and the following day (p = 0.02). Controls had less swollen wounds compared to piglets assigned to treatments M, L and LM (p < 0.001). The proportion of piglets with high SAA concentration (over threshold values 200, 400 mg/l) was higher (p = 0.005; p = 0.05) for C + L compared to M + LM. Ear temperature was higher (p < 0.01) for controls compared to L and LM. There were no significant treatment effects for skin temperature, weight gain, sickness treatments or mortality. CONCLUSIONS: The study concludes that lidocaine reduced pain during castration and that meloxicam reduced pain after castration. The study also concludes that the herdsmen were able to administer local anaesthesia effectively. PMID- 21627798 TI - Multiplex single-nucleotide polymorphism typing of the human Y chromosome using TaqMan probes. AB - BACKGROUND: The analysis of human Y-chromosome variation in the context of population genetics and forensics requires the genotyping of dozens to hundreds of selected single-nucleotide polymorphisms (SNPs). In the present study, we developed a 121-plex (121 SNPs in a single array) TaqMan array capable of distinguishing most haplogroups and subhaplogroups on the Y-chromosome human phylogeny in Europe. RESULTS: We present data from 264 samples from several European areas and ethnic groups. The array developed in this study shows >99% accuracy of assignation to the Y human phylogeny (with an average call rate of genotypes >96%). CONCLUSIONS: We have created and evaluated a robust and accurate Y-chromosome multiplex which minimises the possible errors due to mixup when typing the same sample in several independent reactions. PMID- 21627799 TI - Identification of a functional docking site in the Rpn1 LRR domain for the UBA UBL domain protein Ddi1. AB - BACKGROUND: The proteasome is a multi-subunit protein machine that is the final destination for cellular proteins that have been marked for degradation via an ubiquitin (Ub) chain appendage. These ubiquitylated proteins either bind directly to the intrinsic proteasome ubiqutin chain receptors Rpn10, Rpn13, or Rpt5, or are shuttled to the proteasome by Rad23, Dsk2, or Ddi1. The latter proteins share an Ub association domain (UBA) for binding poly-Ub chains and an Ub-like-domain (UBL) for binding to the proteasome. It has been proposed that shuttling receptors dock on the proteasome via Rpn1, but the precise nature of the docking site remains poorly defined. RESULTS: To shed light on the recruitment of shuttling receptors to the proteasome, we performed both site-directed mutagenesis and genetic screening to identify mutations in Rpn1 that disrupt its binding to UBA-UBL proteins. Here we demonstrate that delivery of Ub conjugates and docking of Ddi1 (and to a lesser extent Dsk2) to the proteasome are strongly impaired by an aspartic acid to alanine point mutation in the highly-conserved D517 residue of Rpn1. Moreover, degradation of the Ddi1-dependent proteasome substrate, Ufo1, is blocked in rpn1-D517A yeast cells. By contrast, Rad23 recruitment to the proteasome is not affected by rpn1-D517A. CONCLUSIONS: These studies provide insight into the mechanism by which the UBA-UBL protein Ddi1 is recruited to the proteasome to enable Ub-dependent degradation of its ligands. Our studies suggest that different UBA-UBL proteins are recruited to the proteasome by distinct mechanisms. PMID- 21627800 TI - The development and characterization of a 60K SNP chip for chicken. AB - BACKGROUND: In livestock species like the chicken, high throughput single nucleotide polymorphism (SNP) genotyping assays are increasingly being used for whole genome association studies and as a tool in breeding (referred to as genomic selection). To be of value in a wide variety of breeds and populations, the success rate of the SNP genotyping assay, the distribution of the SNP across the genome and the minor allele frequencies (MAF) of the SNPs used are extremely important. RESULTS: We describe the design of a moderate density (60k) Illumina SNP BeadChip in chicken consisting of SNPs known to be segregating at high to medium minor allele frequencies (MAF) in the two major types of commercial chicken (broilers and layers). This was achieved by the identification of 352,303 SNPs with moderate to high MAF in 2 broilers and 2 layer lines using Illumina sequencing on reduced representation libraries. To further increase the utility of the chip, we also identified SNPs on sequences currently not covered by the chicken genome assembly (Gallus_gallus-2.1). This was achieved by 454 sequencing of the chicken genome at a depth of 12x and the identification of SNPs on 454 derived contigs not covered by the current chicken genome assembly. In total we added 790 SNPs that mapped to 454-derived contigs as well as 421 SNPs with a position on Chr_random of the current assembly. The SNP chip contains 57,636 SNPs of which 54,293 could be genotyped and were shown to be segregating in chicken populations. Our SNP identification procedure appeared to be highly reliable and the overall validation rate of the SNPs on the chip was 94%. We were able to map 328 SNPs derived from the 454 sequence contigs on the chicken genome. The majority of these SNPs map to chromosomes that are already represented in genome build Gallus_gallus-2.1.0. Twenty-eight SNPs were used to construct two new linkage groups most likely representing two micro-chromosomes not covered by the current genome assembly. CONCLUSIONS: The high success rate of the SNPs on the Illumina chicken 60K Beadchip emphasizes the power of Next generation sequence (NGS) technology for the SNP identification and selection step. The identification of SNPs from sequence contigs derived from NGS sequencing resulted in improved coverage of the chicken genome and the construction of two new linkage groups most likely representing two chicken micro-chromosomes. PMID- 21627801 TI - Global analysis of gene expression in response to L-Cysteine deprivation in the anaerobic protozoan parasite Entamoeba histolytica. AB - BACKGROUND: Entamoeba histolytica, an enteric protozoan parasite, causes amebic colitis and extra intestinal abscesses in millions of inhabitants of endemic areas. E. histolytica completely lacks glutathione metabolism but possesses L cysteine as the principle low molecular weight thiol. L-Cysteine is essential for the structure, stability, and various protein functions, including catalysis, electron transfer, redox regulation, nitrogen fixation, and sensing for regulatory processes. Recently, we demonstrated that in E. histolytica, L cysteine regulates various metabolic pathways including energy, amino acid, and phospholipid metabolism. RESULTS: In this study, employing custom-made Affymetrix microarrays, we performed time course (3, 6, 12, 24, and 48 h) gene expression analysis upon L-cysteine deprivation. We identified that out of 9,327 genes represented on the array, 290 genes encoding proteins with functions in metabolism, signalling, DNA/RNA regulation, electron transport, stress response, membrane transport, vesicular trafficking/secretion, and cytoskeleton were differentially expressed (>=3 fold) at one or more time points upon L-cysteine deprivation. Approximately 60% of these modulated genes encoded proteins of no known function and annotated as hypothetical proteins. We also attempted further functional analysis of some of the most highly modulated genes by L-cysteine depletion. CONCLUSIONS: To our surprise, L-cysteine depletion caused only limited changes in the expression of genes involved in sulfur-containing amino acid metabolism and oxidative stress defense. In contrast, we observed significant changes in the expression of several genes encoding iron sulfur flavoproteins, a major facilitator super-family transporter, regulator of nonsense transcripts, NADPH-dependent oxido-reductase, short chain dehydrogenase, acetyltransferases, and various other genes involved in diverse cellular functions. This study represents the first genome-wide analysis of transcriptional changes induced by L cysteine deprivation in protozoan parasites, and in eukaryotic organisms where L cysteine represents the major intracellular thiol. PMID- 21627802 TI - MicroRNA profiling of rhesus macaque embryonic stem cells. AB - BACKGROUND: MicroRNAs (miRNAs) play important roles in embryonic stem cell (ESC) self-renewal and pluripotency. Numerous studies have revealed human and mouse ESC miRNA profiles. As a model for human-related study, the rhesus macaque is ideal for delineating the regulatory mechanisms of miRNAs in ESCs. However, studies on rhesus macaque (r)ESCs are lacking due to limited rESC availability and a need for systematic analyses of fundamental rESC characteristics. RESULTS: We established three rESC lines and profiled microRNA using Solexa sequencing resulting in 304 known and 66 novel miRNAs. MiRNA profiles were highly conserved between rESC lines and predicted target genes were significantly enriched in differentiation pathways. Further analysis of the miRNA-target network indicated that gene expression regulated by miRNAs was negatively correlated to their evolutionary rate in rESCs. Moreover, a cross-species comparison revealed an overall conservation of miRNA expression patterns between human, mouse and rhesus macaque ESCs. However, we identified three miRNA clusters (miR-467, the miRNA cluster in the imprinted Dlk1-Dio3 region and C19MC) that showed clear interspecies differences. CONCLUSIONS: rESCs share a unique miRNA set that may play critical roles in self-renewal and pluripotency. MiRNA expression patterns are generally conserved between species. However, species and/or lineage specific miRNA regulation changed during evolution. PMID- 21627804 TI - Aiming for the complete utilization of sugar-beet pulp: Examination of the effects of mild acid and hydrothermal pretreatment followed by enzymatic digestion. AB - BACKGROUND: Biomass use for the production of bioethanol or platform chemicals requires efficient breakdown of biomass to fermentable monosaccharides. Lignocellulosic feedstocks often require physicochemical pretreatment before enzymatic hydrolysis can begin. The optimal pretreatment can be different for different feedstocks, and should not lead to biomass destruction or formation of toxic products. METHODS: We examined the influence of six mild sulfuric acid or water pretreatments at different temperatures on the enzymatic degradability of sugar-beet pulp (SBP). RESULTS: We found that optimal pretreatment at 140 degrees C of 15 minutes in water was able to solubilize 60% w/w of the total carbohydrates present, mainly pectins. More severe treatments led to the destruction of the solubilized sugars, and the subsequent production of the sugar degradation products furfural, hydroxymethylfurfural, acetic acid and formic acid. The pretreated samples were successfully degraded enzymatically with an experimental cellulase preparation. CONCLUSIONS: In this study, we found that pretreatment of SBP greatly facilitated the subsequent enzymatic degradation within economically feasible time ranges and enzyme levels. In addition, pretreatment of SBP can be useful to fractionate functional ingredients such as arabinans and pectins from cellulose. We found that the optimal combined severity factor to enhance the enzymatic degradation of SBP was between log R'0 = -2.0 and log R'0 = -1.5. The optimal pretreatment and enzyme treatment solubilized up to 80% of all sugars present in the SBP, including >=90% of the cellulose. PMID- 21627803 TI - Seroprevalence, predictors and estimated incidence of maternal and neonatal Herpes Simplex Virus type 2 infection in semi-urban women in Kilifi, Kenya. AB - BACKGROUND: Herpes Simplex Virus type 2 (HSV-2) has public health importance as a leading cause of genital ulcers, a co-factor in HIV-1 acquisition and transmission and as a cause of neonatal herpes infections. Little is known of its epidemiology and burden in Coastal Kenya. METHODS: We screened plasma samples for HSV-2 infection from 826 women aged 15-34 years who participated in an HIV-1 survey in Kilifi in 2004. The sample comprised 563 women selected randomly from a demographic surveillance system (DSS) and 263 women who presented for voluntary counseling and testing (VCT). Predictors for HSV-2 seropositivity were determined using multivariate logistic regression. The incidence of HSV-2 infection and risk of neonatal herpes were estimated by a simple catalytic model fitted to age seroprevalence data. RESULTS: HSV-2 prevalence was 32% in the DSS recruits vs. 44% in the VCT recruits (P < 0.001), while, HIV-1 prevalence was 8% in the DSS recruits vs. 12% in the VCT recruits (P = 0.12). Independent risk factors for HSV 2 infection in all women were: older age (30-34 years; odds ratio (OR) 10.5, 95% confidence interval (CI): 5.2 - 21.0), recruitment from VCT (OR 1.5, 95% CI: 1.1 2.1), history of genital ulcers (OR 1.7, 95% CI: 1.2 - 2.3) and HIV infection (OR 2.7, 95% CI: 1.6-4.6). Education beyond primary (OR 0.7, 95% CI: 0.5 - 0.9) was inversely associated with HSV-2 infection. In the DSS sample, HSV-2 incidence was estimated at 4 cases (95% CI: 3.3 - 4.4) per 100 women per year, 17 cases (95% CI: 16-18) per 1,000 pregnancies per year and 33 neonatal cases (95% CI: 31 36) per 100,000 births per year. CONCLUSIONS: HSV-2 transmission is rapid following the onset of sexual activity and likely to result in a significant burden of genital ulcer disease. Nevertheless, the burden of neonatal HSV-2 can be predicted to be low. Educating young women about HSV-2 infection may help in reducing its burden in this semi-urban population. PMID- 21627805 TI - Song exposure regulates known and novel microRNAs in the zebra finch auditory forebrain. AB - BACKGROUND: In an important model for neuroscience, songbirds learn to discriminate songs they hear during tape-recorded playbacks, as demonstrated by song-specific habituation of both behavioral and neurogenomic responses in the auditory forebrain. We hypothesized that microRNAs (miRNAs or miRs) may participate in the changing pattern of gene expression induced by song exposure. To test this, we used massively parallel Illumina sequencing to analyse small RNAs from auditory forebrain of adult zebra finches exposed to tape-recorded birdsong or silence. RESULTS: In the auditory forebrain, we identified 121 known miRNAs conserved in other vertebrates. We also identified 34 novel miRNAs that do not align to human or chicken genomes. Five conserved miRNAs showed significant and consistent changes in copy number after song exposure across three biological replications of the song-silence comparison, with two increasing (tgu-miR-25, tgu miR-192) and three decreasing (tgu-miR-92, tgu-miR-124, tgu-miR-129-5p). We also detected a locus on the Z sex chromosome that produces three different novel miRNAs, with supporting evidence from Northern blot and TaqMan qPCR assays for differential expression in males and females and in response to song playbacks. One of these, tgu-miR-2954-3p, is predicted (by TargetScan) to regulate eight song-responsive mRNAs that all have functions in cellular proliferation and neuronal differentiation. CONCLUSIONS: The experience of hearing another bird singing alters the profile of miRNAs in the auditory forebrain of zebra finches. The response involves both known conserved miRNAs and novel miRNAs described so far only in the zebra finch, including a novel sex-linked, song-responsive miRNA. These results indicate that miRNAs are likely to contribute to the unique behavioural biology of learned song communication in songbirds. PMID- 21627806 TI - Evidence of association between nucleosome occupancy and the evolution of transcription factor binding sites in yeast. AB - BACKGROUND: Divergence of transcription factor binding sites is considered to be an important source of regulatory evolution. The associations between transcription factor binding sites and phenotypic diversity have been investigated in many model organisms. However, the understanding of other factors that contribute to it is still limited. Recent studies have elucidated the effect of chromatin structure on molecular evolution of genomic DNA. Though the profound impact of nucleosome positions on gene regulation has been reported, their influence on transcriptional evolution is still less explored. With the availability of genome-wide nucleosome map in yeast species, it is thus desirable to investigate their impact on transcription factor binding site evolution. Here, we present a comprehensive analysis of the role of nucleosome positioning in the evolution of transcription factor binding sites. RESULTS: We compared the transcription factor binding site frequency in nucleosome occupied regions and nucleosome depleted regions in promoters of old (orthologs among Saccharomycetaceae) and young (Saccharomyces specific) genes; and in duplicate gene pairs. We demonstrated that nucleosome occupied regions accommodate greater binding site variations than nucleosome depleted regions in young genes and in duplicate genes. This finding was confirmed by measuring the difference in evolutionary rates of binding sites in sensu stricto yeasts at nucleosome occupied regions and nucleosome depleted regions. The binding sites at nucleosome occupied regions exhibited a consistently higher evolution rate than those at nucleosome depleted regions, corroborating the difference in the selection constraints at the two regions. Finally, through site-directed mutagenesis experiment, we found that binding site gain or loss events at nucleosome depleted regions may cause more expression differences than those in nucleosome occupied regions. CONCLUSIONS: Our study indicates the existence of different selection constraint on binding sites at nucleosome occupied regions than at the nucleosome depleted regions. We found that the binding sites have a different rate of evolution at nucleosome occupied and depleted regions. Finally, using transcription factor binding site-directed mutagenesis experiment, we confirmed the difference in the impact of binding site changes on expression at these regions. Thus, our work demonstrates the importance of composite analysis of chromatin and transcriptional evolution. PMID- 21627807 TI - Examining mindfulness-based stress reduction: perceptions from minority older adults residing in a low-income housing facility. AB - BACKGROUND: Mindfulness-based stress reduction (MBSR) programs are becoming increasingly common, but have not been studied in low income minority older populations. We sought to understand which parts of MBSR were most important to practicing MBSR members of this population, and to understand whether they apply their training to daily challenges. METHODS: We conducted three focus groups with 13 current members of an MBSR program. Participants were African American women over the age of 60 in a low-income housing residence. We tape recorded each session and subsequently used inductive content analysis to identify primary themes. RESULTS AND DISCUSSION: Analysis of the focus group responses revealed three primary themes stress management, applying mindfulness, and the social support of the group meditation. The stressors they cited using MBSR with included growing older with physical pain, medical tests, financial strain, and having grandchildren with significant mental, physical, financial or legal hardships. We found that participants particularly used their MBSR training for coping with medical procedures, and managing both depression and anger. CONCLUSION: A reflective stationary intervention delivered in-residence could be an ideal mechanism to decrease stress in low-income older adult's lives and improve their health. PMID- 21627808 TI - Health service utilization in IBD: comparison of self-report and administrative data. AB - BACKGROUND: The reliability of self-report regarding health care utilization in inflammatory bowel disease (IBD) is unknown. If proven reliable, it could help justify self-report as a means of determining health care utilization and associated costs. METHODS: The Manitoba IBD Cohort Study is a population-based longitudinal study of participants diagnosed within 7 years of enrollment. Health care utilization was assessed through standardized interview. Participants (n = 352) reported the total number of nights hospitalized, frequency of physician contacts in the prior 12 months and whether the medical contacts were for IBD related reasons or not. Reports of recent antibiotic use were also recorded. Actual utilization was drawn from the administrative database of Manitoba Health, the single comprehensive provincial health insurer. RESULTS: According to the administrative data, 15% of respondents had an overnight hospitalization, while 10% had an IBD-related hospitalization. Self-report concordance was highly sensitive (92%; 82%) and specific (96%; 97%, respectively). 97% of participants had contact with a physician in the previous year, and 69% had IBD-related visits. Physician visits were significantly under-reported and there was a trend to over-report the number of nights in hospital. CONCLUSIONS: Self-report data can be helpful in evaluating health service utilization, provided that the researcher is aware of the systematic sources of bias. Outpatient visits are well identified by self-report. The discordance for the type of outpatient visit may be either a weakness of self-report or a flaw in diagnosis coding of the administrative data. If administrative data are not available, self-report information may be a cost-effective alternative, particularly for hospitalizations. PMID- 21627809 TI - A color spectrographic phonocardiography (CSP) applied to the detection and characterization of heart murmurs: preliminary results. AB - BACKGROUND: Although cardiac auscultation remains important to detect abnormal sounds and murmurs indicative of cardiac pathology, the application of electronic methods remains seldom used in everyday clinical practice. In this report we provide preliminary data showing how the phonocardiogram can be analyzed using color spectrographic techniques and discuss how such information may be of future value for noninvasive cardiac monitoring. METHODS: We digitally recorded the phonocardiogram using a high-speed USB interface and the program Gold Wave http://www.goldwave.com in 55 infants and adults with cardiac structural disease as well as from normal individuals and individuals with innocent murmurs. Color spectrographic analysis of the signal was performed using Spectrogram (Version 16) as a well as custom MATLAB code. RESULTS: Our preliminary data is presented as a series of seven cases. CONCLUSIONS: We expect the application of spectrographic techniques to phonocardiography to grow substantially as ongoing research demonstrates its utility in various clinical settings. Our evaluation of a simple, low-cost phonocardiographic recording and analysis system to assist in determining the characteristic features of heart murmurs shows promise in helping distinguish innocent systolic murmurs from pathological murmurs in children and is expected to useful in other clinical settings as well. PMID- 21627811 TI - Diagnostic dilemmas of squamous differentiation in prostate carcinoma case report and review of the literature. AB - We report a case of pure squamous cell carcinoma involving the prostate and urinary bladder and describe the diagnostic dilemmas that we faced in trying to determine its origin. The patient was diagnosed ten years ago with prostatic adenocarcinoma treated with radioactive seed implantation. During the last year he also underwent a TURP procedure for urinary obstruction complicated by multiple infections. Postsurgery, the patient developed colo-urethral fistula and decision to perform cystoprostatectomy was taken. Excision illustrated a tumor mass replacing the entire prostate that microscopically proved to be squamous cell carcinoma. The challenge that we encountered was to determine its origin, the possibilities being divergent differentiation from adenocarcinoma post radiation therapy, de novo neoplasm or urothelial carcinoma with extensive squamous differentiation. Our literature review showed also that the etiology of prostatic squamous carcinoma is still unclear. We present our approach in an attempt to solve this dilemma. PMID- 21627810 TI - Estrogen and progesterone-related gene variants and colorectal cancer risk in women. AB - BACKGROUND: Observational studies and randomized trials have suggested that estrogens and/or progesterone may lower the risk for colorectal cancer. Inherited variation in the sex-hormone genes may be one mechanism by which sex hormones affect colorectal cancer, although data are limited. METHOD: We conducted a comprehensive evaluation of single nucleotide polymorphisms (SNPs) in genes encoding 3 hormone receptors (ESR1, ESR2, PGR) and 5 hormone synthesizers (CYP19A1 and CYP17A1, HSD17B1, HSD17B2, HSD17B4) among 427 women with incident colorectal cancer and 871 matched controls who were Caucasians of European ancestry from 93676 postmenopausal women enrolled in the Women's Health Initiative Observational cohort. A total of 242 haplotype-tagging and functional SNPs in the 8 genes were included for analysis. Unconditional logistic regression with adjustment for age and hysterectomy status was used to estimate odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: We observed a weak association between the CYP17A1 rs17724534 SNP and colorectal cancer risk (OR per risk allele (A) = 1.39, 95% CI = 1.09-1.78, corrected p-value = 0.07). In addition, a suggestive interaction between rs17724534 and rs10883782 in 2 discrete LD blocks of CYP17A1 was observed in relation to colorectal cancer (empirical p value = 0.04). Moreover, one haplotype block of CYP19A1 was associated with colorectal cancer (corrected global p value = 0.02), which likely reflected the association with the tagging SNP, rs1902584, in the block. CONCLUSION: Our findings offer some support for a suggestive association of CYP17A1 and CYP19A1 variants with colorectal cancer risk. PMID- 21627812 TI - Assisted reproduction: what factors interfere in the professional's decisions? Are single women an issue? AB - BACKGROUND: With the development of medical technology, many countries around the world have been implementing ethical guidelines and laws regarding Medically Assisted Reproduction (MAR). A physician's reproductive decisions are not solely based on technical criteria but are also influenced by society values. Therefore, the aim of this study was to analyze the factors prioritized by MAR professionals when deciding on whether to accept to perform assisted reproduction and to show any existing cultural differences. METHODS: Cross-sectional study involving 224 healthcare professionals working with assisted reproduction in Brazil, Italy, Germany and Greece. Instrument used for data collection: a questionnaire, followed by the description of four special MAR cases (a single woman, a lesbian couple, an HIV discordant couple and gender selection) which included case specific questions regarding the professionals' decision on whether to perform the requested procedure as well as the following factors: socio-demographic variables, moral and legal values as well as the technical aspects which influence decision-making. RESULTS: Only the case involving a single woman who wishes to have a child (without the intention of having a partner in the future) demonstrated significant differences. Therefore, the study was driven towards the results of this case specifically. The analyses we performed demonstrated that professionals holding a Master's Degree, those younger in age, female professionals, those having worked for less time in reproduction, those in private clinics and Brazilian health professionals all had a greater tendency to perform the procedure in that case. A multivariate analysis demonstrated that the reasons for the professional's decision to perform the procedure were the woman's right to gestate and the duty of MAR professionals to help her. The professionals who decided not to perform the procedure identified the woman's marital status and the child's right to a father as the reason to withhold treatment. CONCLUSION: The study indicates differences among countries in the evaluation of the single woman case. It also discloses the undervaluation of bioethics committees and the need for a greater participation of healthcare professionals in debates on assisted reproduction laws. PMID- 21627813 TI - Regional expression of HOXA4 along the aorta and its potential role in human abdominal aortic aneurysms. AB - BACKGROUND: The infrarenal abdominal aorta exhibits increased disease susceptibility relative to other aortic regions. Allograft studies exchanging thoracic and abdominal segments showed that regional susceptibility is maintained regardless of location, suggesting substantial roles for embryological origin, tissue composition and site-specific gene expression. RESULTS: We analyzed gene expression with microarrays in baboon aortas, and found that members of the HOX gene family exhibited spatial expression differences. HOXA4 was chosen for further study, since it had decreased expression in the abdominal compared to the thoracic aorta. Western blot analysis from 24 human aortas demonstrated significantly higher HOXA4 protein levels in thoracic compared to abdominal tissues (P < 0.001). Immunohistochemical staining for HOXA4 showed nuclear and perinuclear staining in endothelial and smooth muscle cells in aorta. The HOXA4 transcript levels were significantly decreased in human abdominal aortic aneurysms (AAAs) compared to age-matched non-aneurysmal controls (P < 0.00004). Cultured human aortic endothelial and smooth muscle cells stimulated with INF gamma (an important inflammatory cytokine in AAA pathogenesis) showed decreased levels of HOXA4 protein (P < 0.0007). CONCLUSIONS: Our results demonstrated spatial variation in expression of HOXA4 in human aortas that persisted into adulthood and that downregulation of HOXA4 expression was associated with AAAs, an important aortic disease of the ageing population. PMID- 21627814 TI - Optimizing the diagnostic power with gastric emptying scintigraphy at multiple time points. AB - BACKGROUND: Gastric Emptying Scintigraphy (GES) at intervals over 4 hours after a standardized radio-labeled meal is commonly regarded as the gold standard for diagnosing gastroparesis. The objectives of this study were: 1) to investigate the best time point and the best combination of multiple time points for diagnosing gastroparesis with repeated GES measures, and 2) to contrast and cross validate Fisher's Linear Discriminant Analysis (LDA), a rank based Distribution Free (DF) approach, and the Classification And Regression Tree (CART) model. METHODS: A total of 320 patients with GES measures at 1, 2, 3, and 4 hour (h) after a standard meal using a standardized method were retrospectively collected. Area under the Receiver Operating Characteristic (ROC) curve and the rate of false classification through jackknife cross-validation were used for model comparison. RESULTS: Due to strong correlation and an abnormality in data distribution, no substantial improvement in diagnostic power was found with the best linear combination by LDA approach even with data transformation. With DF method, the linear combination of 4-h and 3-h increased the Area Under the Curve (AUC) and decreased the number of false classifications (0.87; 15.0%) over individual time points (0.83, 0.82; 15.6%, 25.3%, for 4-h and 3-h, respectively) at a higher sensitivity level (sensitivity=0.9). The CART model using 4 hourly GES measurements along with patient's age was the most accurate diagnostic tool (AUC=0.88, false classification=13.8%). Patients having a 4-h gastric retention value>10% were 5 times more likely to have gastroparesis (179/207=86.5%) than those with <=10% (18/113=15.9%). CONCLUSIONS: With a mixed group of patients either referred with suspected gastroparesis or investigated for other reasons, the CART model is more robust than the LDA and DF approaches, capable of accommodating covariate effects and can be generalized for cross institutional applications, but could be unstable if sample size is limited. PMID- 21627815 TI - The 5S rDNA family evolves through concerted and birth-and-death evolution in fish genomes: an example from freshwater stingrays. AB - BACKGROUND: Ribosomal 5S genes are well known for the critical role they play in ribosome folding and functionality. These genes are thought to evolve in a concerted fashion, with high rates of homogenization of gene copies. However, the majority of previous analyses regarding the evolutionary process of rDNA repeats were conducted in invertebrates and plants. Studies have also been conducted on vertebrates, but these analyses were usually restricted to the 18S, 5.8S and 28S rRNA genes. The recent identification of divergent 5S rRNA gene paralogs in the genomes of elasmobranches and teleost fishes indicate that the eukaryotic 5S rRNA gene family has a more complex genomic organization than previously thought. The availability of new sequence data from lower vertebrates such as teleosts and elasmobranches enables an enhanced evolutionary characterization of 5S rDNA among vertebrates. RESULTS: We identified two variant classes of 5S rDNA sequences in the genomes of Potamotrygonidae stingrays, similar to the genomes of other vertebrates. One class of 5S rRNA genes was shared only by elasmobranches. A broad comparative survey among 100 vertebrate species suggests that the 5S rRNA gene variants in fishes originated from rounds of genome duplication. These variants were then maintained or eliminated by birth-and-death mechanisms, under intense purifying selection. Clustered multiple copies of 5S rDNA variants could have arisen due to unequal crossing over mechanisms. Simultaneously, the distinct genome clusters were independently homogenized, resulting in the maintenance of clusters of highly similar repeats through concerted evolution. CONCLUSIONS: We believe that 5S rDNA molecular evolution in fish genomes is driven by a mixed mechanism that integrates birth-and-death and concerted evolution. PMID- 21627816 TI - Peanut sensitization in a group of allergic Egyptian children. AB - BACKGROUND: There are no published data on peanut sensitization in Egypt and the problem of peanut allergy seems underestimated. We sought to screen for peanut sensitization in a group of atopic Egyptian children in relation to their phenotypic manifestations. METHODS: We consecutively enrolled 100 allergic children; 2-10 years old (mean 6.5 yr). The study measurements included clinical evaluation for site of allergy, possible precipitating factors, consumption of peanuts (starting age and last consumption), duration of breast feeding, current treatment, and family history of allergy as well as skin prick testing with a commercial peanut extract, and serum peanut specific and total IgE estimation. Children who were found sensitized to peanuts were subjected to an open oral peanut challenge test taking all necessary precautions. RESULTS: Seven subjects (7%) were sensitized and three out of six of them had positive oral challenge denoting allergy to peanuts. The sensitization rates did not vary significantly with gender, age, family history of allergy, breast feeding duration, clinical form of allergy, serum total IgE, or absolute eosinophil count. All peanut sensitive subjects had skin with or without respiratory allergy. CONCLUSIONS: Peanut allergy does not seem to be rare in atopic children in Egypt. Skin prick and specific IgE testing are effective screening tools to determine candidates for peanut oral challenging. Wider scale multicenter population-based studies are needed to assess the prevalence of peanut allergy and its clinical correlates in our country. PMID- 21627817 TI - Characteristics of patients with venous thromboembolism and atrial fibrillation in Venezuela. AB - BACKGROUND: Studies describing venous thromboembolic event (VTEE) and atrial fibrillation (AF) in South American populations are limited. The aim of this cross-sectional study was to describe the characteristics of Venezuelan patients admitted and treated for these conditions. METHODS: A retrospective medical record review of 1397 consecutive patients admitted to three private hospitals or clinics between January 2000 and December 2005 was performed. Data was collected on demographics, anthropometrics, hospital visit, comorbidities and treatment. RESULTS: Among 401 VTEE and 996 AF patients, men were more likely to have AF (58%) while more women experienced a VTEE (58%). Most patients were admitted via the emergency room (87%) and had only one event during the study period (83%). Common comorbidities included hypertension (46%), heart failure (17%), diabetes (12%) and congestive heart failure (11%). Characteristics of Venezuelan patients with VTEE and AF are similar to that reported in the literature for other populations. CONCLUSIONS: These results provide background characteristics for future studies assessing risk factors for AF and VTEE in South American populations. PMID- 21627818 TI - Social inequalities, regional disparities and health inequity in North African countries. AB - BACKGROUND: During the last decades, North African countries have substantially improved economic, social and health conditions of their populations in average. In all countries, human development in general and life expectancy, literacy and per capita income in particular have increased. However, improvement was not equally shared between groups of different milieu, regions or level of income. Social inequalities and health inequity have persisted or even worsened. Data are generally scarce and few studies were devoted to this topic in North Africa as a region. In this paper, we carry out a comparative study on the achievements of these countries, not only in terms of human development and its components but also in terms of inequalities' reduction and health equity. METHOD: This study is based on data available for comparison between North African countries. The main data sources are provided by reports released by the World Health Organisation (WHO), United Nations Development Programme (UNDP), United Nations Children's Fund (UNICEF), the World Bank, surveys such as Demographic Health Surveys (DHS) and Multiple Indicator Cluster Surveys (MICS) and finally recent papers published on equity in different countries of the region. RESULTS AND DISCUSSION: There is no doubt that education, health and human development in general have improved in North Africa during the last decades. Improvement was, however, uneven and unequally enjoyed by different socioeconomic groups. Indeed, each country included in this study shows large urban-rural disparities, discrepancies between advantaged and disadvantaged regions and cities; and unacceptable differences between rich and poor. Health inequity is particularly seen through access to health services and infant mortality. CONCLUSION: During the last decades, North African decision makers have endeavoured to improve social and economic conditions of their populations. Globally, health, education and living standard in general have substantially improved in average. However, North African countries have still a long way to go to reduce social inequalities and health inequity at different levels: rural-urban, advantaged-marginalised regions and cities, between groups of different level of income and wealth. The challenge for the next decade is not only to improve economic, social and health conditions in average but also and mainly to reduce avoidable inequalities in parallel. PMID- 21627819 TI - Managing resources in NHS dentistry: using health economics to inform commissioning decisions. AB - BACKGROUND: The aim of this study is to develop, apply and evaluate an economics based framework to assist commissioners in their management of finite resources for local dental services. In April 2006, Primary Care Trusts in England were charged with managing finite dental budgets for the first time, yet several independent reports have since criticised the variability in commissioning skills within these organisations. The study will explore the views of stakeholders (dentists, patients and commissioners) regarding priority setting and the criteria used for decision-making and resource allocation. Two inter-related case studies will explore the dental commissioning and resource allocation processes through the application of a pragmatic economics-based framework known as Programme Budgeting and Marginal Analysis. METHODS/DESIGN: The study will adopt an action research approach. Qualitative methods including semi-structured interviews, focus groups, field notes and document analysis will record the views of participants and their involvement in the research process. The first case study will be based within a Primary Care Trust where mixed methods will record the views of dentists, patients and dental commissioners on issues, priorities and processes associated with managing local dental services. A Programme Budgeting and Marginal Analysis framework will be applied to determine the potential value of economic principles to the decision-making process. A further case study will be conducted in a secondary care dental teaching hospital using the same approach. Qualitative data will be analysed using thematic analysis and managed using a framework approach. DISCUSSION: The recent announcement by government regarding the proposed abolition of Primary Care Trusts may pose challenges for the research team regarding their engagement with the research study. However, whichever commissioning organisations are responsible for resource allocation for dental services in the future; resource scarcity is highly likely to remain an issue. Wider understanding of the complexities of priority setting and resource allocation at local levels are important considerations in the development of dental commissioning processes, national oral health policy and the future new dental contract which is expected to be implemented in April 2014. PMID- 21627820 TI - BMP signaling components in embryonic transcriptomes of the hover fly Episyrphus balteatus (Syrphidae). AB - BACKGROUND: In animals, signaling of Bone Morphogenetic Proteins (BMPs) is essential for dorsoventral (DV) patterning of the embryo, but how BMP signaling evolved with changes in embryonic DV differentiation is largely unclear. Based on the extensive knowledge of BMP signaling in Drosophila melanogaster, the morphological diversity of extraembryonic tissues in different fly species provides a comparative system to address this question. The closest relatives of D. melanogaster with clearly distinct DV differentiation are hover flies (Diptera: Syrphidae). The syrphid Episyrphus balteatus is a commercial bio-agent against aphids and has been established as a model organism for developmental studies and chemical ecology. The dorsal blastoderm of E. balteatus gives rise to two extraembryonic tissues (serosa and amnion), whereas in D. melanogaster, the dorsal blastoderm differentiates into a single extraembryonic epithelium (amnioserosa). Recent studies indicate that several BMP signaling components of D. melanogaster, including the BMP ligand Screw (Scw) and other extracellular regulators, evolved in the dipteran lineage through gene duplication and functional divergence. These findings raise the question of whether the complement of BMP signaling components changed with the origin of the amnioserosa. RESULTS: To search for BMP signaling components in E. balteatus, we generated and analyzed transcriptomes of freshly laid eggs (0-30 minutes) and late blastoderm to early germband extension stages (3-6 hours) using Roche/454 sequencing. We identified putative E. balteatus orthologues of 43% of all annotated D. melanogaster genes, including the genes of all BMP ligands and other BMP signaling components. CONCLUSION: The diversification of several BMP signaling components in the dipteran linage of D. melanogaster preceded the origin of the amnioserosa.[Transcriptome sequence data from this study have been deposited at the NCBI Sequence Read Archive (SRP005289); individually assembled sequences have been deposited at GenBank (JN006969-JN006986).]. PMID- 21627821 TI - Purification of phage display-modified bacteriophage T4 by affinity chromatography. AB - BACKGROUND: Affinity chromatography is one of the most efficient protein purification strategies. This technique comprises a one-step procedure with a purification level in the order of several thousand-fold, adaptable for various proteins, differentiated in their size, shape, charge, and other properties. The aim of this work was to verify the possibility of applying affinity chromatography in bacteriophage purification, with the perspective of therapeutic purposes. T4 is a large, icosahedral phage that may serve as an efficient display platform for foreign peptides or proteins. Here we propose a new method of T4 phage purification by affinity chromatography after its modification with affinity tags (GST and Histag) by in vivo phage display. As any permanent introduction of extraneous DNA into a phage genome is strongly unfavourable for medical purposes, integration of foreign motifs with the phage genome was not applied. The phage was propagated in bacteria expressing fusions of the phage protein Hoc with affinity tags from bacterial plasmids, independently from the phage expression system. RESULTS: Elution profiles of phages modified with the specific affinity motifs (compared to non-specific phages) document their binding to the affinity resins and effective elution with standard competitive agents. Non-specific binding was also observed, but was 102-105 times weaker than the specific one. GST-modified bacteriophages were also effectively released from glutathione Sepharose by proteolytic cleavage. The possibility of proteolytic release was designed at the stage of expression vector construction. Decrease in LPS content in phage preparations was dependent on the washing intensity; intensive washing resulted in preparations of 11-40 EU/ml. CONCLUSIONS: Affinity tags can be successfully incorporated into the T4 phage capsid by the in vivo phage display technique and they strongly elevate bacteriophage affinity to a specific resin. Affinity chromatography can be considered as a new phage purification method, appropriate for further investigations and development. PMID- 21627822 TI - An upper limit for macromolecular crowding effects. AB - BACKGROUND: Solutions containing high macromolecule concentrations are predicted to affect a number of protein properties compared to those properties in dilute solution. In cells, these macromolecular crowders have a large range of sizes and can occupy 30% or more of the available volume. We chose to study the stability and ps-ns internal dynamics of a globular protein whose radius is ~2 nm when crowded by a synthetic microgel composed of poly(N-isopropylacrylamide-co-acrylic acid) with particle radii of ~300 nm. RESULTS: Our studies revealed no change in protein rotational or ps-ns backbone dynamics and only mild (~0.5 kcal/mol at 37 degrees C, pH 5.4) stabilization at a volume occupancy of 70%, which approaches the occupancy of closely packing spheres. The lack of change in rotational dynamics indicates the absence of strong crowder-protein interactions. CONCLUSIONS: Our observations are explained by the large size discrepancy between the protein and crowders and by the internal structure of the microgels, which provide interstitial spaces and internal pores where the protein can exist in a dilute solution-like environment. In summary, microgels that interact weakly with proteins do not strongly influence protein dynamics or stability because these large microgels constitute an upper size limit on crowding effects. PMID- 21627823 TI - Studying feasibility and effects of a two-stage nursing staff training in residential geriatric care using a 30 month mixed-methods design [ISRCTN24344776]. AB - BACKGROUND: Transfer techniques and lifting weights often cause back pain and disorders for nurses in geriatric care. The Kinaesthetics care conception claims to be an alternative, yielding benefits for nurses as well as for clients.Starting a multi-step research program on the effects of Kinaesthetics, we assess the feasibility of a two-stage nursing staff training and a pre-post research design. Using quantitative and qualitative success criteria, we address mobilisation from the bed to a chair and backwards, walking with aid and positioning in bed on the staff level as well as on the resident level. In addition, effect estimates should help to decide on and to prepare a controlled trial. METHODS/DESIGN: Standard basic and advanced Kinaesthetics courses (each comprising four subsequent days and an additional counselling day during the following four months) are offered to n = 36 out of 60 nurses in a residential geriatric care home, who are in charge of 76 residents. N = 22 residents needing movement support are participating to this study.On the staff level, measurements include focus group discussions, questionnaires, physical strain self-assessment (Borg scale), video recordings and external observation of patient assistance skills using a specialised instrument (SOPMAS). Questionnaires used on the resident level include safety, comfort, pain, and level of own participation during mobilisation. A functional mobility profile is assessed using a specialised test procedure (MOTPA).Measurements will take place at baseline (T0), after basic training (T1), and after the advanced course (T2). Follow-up focus groups will be offered at T1 and 10 months later (T3). DISCUSSION: Ten criteria for feasibility success are established before the trial, assigned to resources (missing data), processes (drop-out of nurses and residents) and science (minimum effects) criteria. This will help to make rational decision on entering the next stage of the research program. TRIAL REGISTRATION: Current Controlled Trials ISRCTN24344776. PMID- 21627824 TI - Software comparison for evaluating genomic copy number variation for Affymetrix 6.0 SNP array platform. AB - BACKGROUND: Copy number data are routinely being extracted from genome-wide association study chips using a variety of software. We empirically evaluated and compared four freely-available software packages designed for Affymetrix SNP chips to estimate copy number: Affymetrix Power Tools (APT), Aroma.Affymetrix, PennCNV and CRLMM. Our evaluation used 1,418 GENOA samples that were genotyped on the Affymetrix Genome-Wide Human SNP Array 6.0. We compared bias and variance in the locus-level copy number data, the concordance amongst regions of copy number gains/deletions and the false-positive rate amongst deleted segments. RESULTS: APT had median locus-level copy numbers closest to a value of two, whereas PennCNV and Aroma.Affymetrix had the smallest variability associated with the median copy number. Of those evaluated, only PennCNV provides copy number specific quality-control metrics and identified 136 poor CNV samples. Regions of copy number variation (CNV) were detected using the hidden Markov models provided within PennCNV and CRLMM/VanillaIce. PennCNV detected more CNVs than CRLMM/VanillaIce; the median number of CNVs detected per sample was 39 and 30, respectively. PennCNV detected most of the regions that CRLMM/VanillaIce did as well as additional CNV regions. The median concordance between PennCNV and CRLMM/VanillaIce was 47.9% for duplications and 51.5% for deletions. The estimated false-positive rate associated with deletions was similar for PennCNV and CRLMM/VanillaIce. CONCLUSIONS: If the objective is to perform statistical tests on the locus-level copy number data, our empirical results suggest that PennCNV or Aroma.Affymetrix is optimal. If the objective is to perform statistical tests on the summarized segmented data then PennCNV would be preferred over CRLMM/VanillaIce. Specifically, PennCNV allows the analyst to estimate locus-level copy number, perform segmentation and evaluate CNV-specific quality-control metrics within a single software package. PennCNV has relatively small bias, small variability and detects more regions while maintaining a similar estimated false-positive rate as CRLMM/VanillaIce. More generally, we advocate that software developers need to provide guidance with respect to evaluating and choosing optimal settings in order to obtain optimal results for an individual dataset. Until such guidance exists, we recommend trying multiple algorithms, evaluating concordance/discordance and subsequently consider the union of regions for downstream association tests. PMID- 21627825 TI - A novel deconvolution method for modeling UDP-N-acetyl-D-glucosamine biosynthetic pathways based on (13)C mass isotopologue profiles under non-steady-state conditions. AB - BACKGROUND: Stable isotope tracing is a powerful technique for following the fate of individual atoms through metabolic pathways. Measuring isotopic enrichment in metabolites provides quantitative insights into the biosynthetic network and enables flux analysis as a function of external perturbations. NMR and mass spectrometry are the techniques of choice for global profiling of stable isotope labeling patterns in cellular metabolites. However, meaningful biochemical interpretation of the labeling data requires both quantitative analysis and complex modeling. Here, we demonstrate a novel approach that involved acquiring and modeling the timecourses of (13)C isotopologue data for UDP-N-acetyl-D glucosamine (UDP-GlcNAc) synthesized from [U-(13)C]-glucose in human prostate cancer LnCaP-LN3 cells. UDP-GlcNAc is an activated building block for protein glycosylation, which is an important regulatory mechanism in the development of many prominent human diseases including cancer and diabetes. RESULTS: We utilized a stable isotope resolved metabolomics (SIRM) approach to determine the timecourse of (13)Cincorporation from [U-(13)C]-glucose into UDP-GlcNAc in LnCaP LN3 cells. (13)CPositional isotopomers and isotopologues of UDP-GlcNAc were determined by high resolution NMR and Fourier transform-ion cyclotron resonance mass spectrometry. A novel simulated annealing/genetic algorithm, called 'Genetic Algorithm for Isotopologues in Metabolic Systems' (GAIMS) was developed to find the optimal solutions to a set of simultaneous equations that represent the isotopologue compositions, which is a mixture of isotopomer species. The best model was selected based on information theory. The output comprises the timecourse of the individual labeled species, which was deconvoluted into labeled metabolic units, namely glucose, ribose, acetyl and uracil. The performance of the algorithm was demonstrated by validating the computed fractional 13C enrichment in these subunits against experimental data. The reproducibility and robustness of the deconvolution were verified by replicate experiments, extensive statistical analyses, and cross-validation against NMR data. CONCLUSIONS: This computational approach revealed the relative fluxes through the different biosynthetic pathways of UDP-GlcNAc, which comprises simultaneous sequential and parallel reactions, providing new insight into the regulation of UDP-GlcNAc levels and O-linked protein glycosylation. This is the first such analysis of UDP GlcNAc dynamics, and the approach is generally applicable to other complex metabolites comprising distinct metabolic subunits, where sufficient numbers of isotopologues can be unambiguously resolved and accurately measured. PMID- 21627826 TI - Cis-regulation of IRF5 expression is unable to fully account for systemic lupus erythematosus association: analysis of multiple experiments with lymphoblastoid cell lines. AB - INTRODUCTION: Interferon regulatory factor 5 gene (IRF5) polymorphisms are strongly associated with several diseases, including systemic lupus erythematosus (SLE). The association includes risk and protective components. They could be due to combinations of functional polymorphisms and related to cis-regulation of IRF5 expression, but their mechanisms are still uncertain. We hypothesised that thorough testing of the relationships between IRF5 polymorphisms, expression data from multiple experiments and SLE-associated haplotypes might provide useful new information. METHODS: Expression data from four published microarray hybridisation experiments with lymphoblastoid cell lines (57 to 181 cell lines) were retrieved. Genotypes of 109 IRF5 polymorphisms, including four known functional polymorphisms, were considered. The best linear regression models accounting for the IRF5 expression data were selected by using a forward entry procedure. SLE-associated IRF5 haplotypes were correlated with the expression data and with the best cis-regulatory models. RESULTS: A large fraction of variability in IRF5 expression was accounted for by linear regression models with IRF5 polymorphisms, but at a different level in each expression data set. Also, the best models from each expression data set were different, although there was overlap between them. The SNP introducing an early polyadenylation signal, rs10954213, was included in the best models for two of the expression data sets and in good models for the other two data sets. The SLE risk haplotype was associated with high IRF5 expression in the four expression data sets. However, there was also a trend towards high IRF5 expression with some protective and neutral haplotypes, and the protective haplotypes were not associated with IRF5 expression. As a consequence, correlation between the cis-regulatory best models and SLE-associated haplotypes, regarding either the risk or protective component, was poor. CONCLUSIONS: Our analysis indicates that although the SLE risk haplotype of IRF5 is associated with high expression of the gene, cis-regulation of IRF5 expression is not enough to fully account for IRF5 association with SLE susceptibility, which indicates the need to identify additional functional changes in this gene. PMID- 21627827 TI - Notochordal conditioned media from tissue increases proteoglycan accumulation and promotes a healthy nucleus pulposus phenotype in human mesenchymal stem cells. AB - INTRODUCTION: Notochordal cells (NCs) are influential in development of the intervertebral disc (IVD) and species that retain NCs do not degenerate. IVD repair using bone marrow derived mesenchymal stem cells (MSCs) is an attractive approach and the harsh microenvironment of the IVD suggests pre-differentiation is a necessary first step. The goal of this study was to use soluble factors from NCs in alginate and NCs in their native tissue to differentiate human MSCs to a young nucleus pulposus (NP) phenotype. METHODS: Human MSCs (cultured under micromass conditions for 21 days in hypoxia) were differentiated with conditioned medium derived from porcine notochordal cells in native tissue (NCT) or in alginate beads (NCA), and compared with chondrogenic (TGFbeta-3) or basal medium. A PCR array of 42 genes was utilized to screen a large number of genes known to be associated with the healthy NP phenotype and pellet cultures were also evaluated for glycosaminoglycan content, histology and viability. Proteomic analysis was used to assess candidate soluble factors in NCA and NCT. RESULTS: Notochordal cell conditioned media had diverse effects on MSC phenotype. NCT resulted in the highest levels of glycosaminoglycan (GAG), as well as up regulation of SOX9 and Collagen II gene expression. NCA demonstrated effects that were catabolic yet also anti-fibrotic and minimally hypertrophic with down regulation of Collagens I and III and low levels of Collagen X, respectively. Micromass culture and hypoxic conditions were sufficient to promote chondrogenesis demonstrating that both basal and chondrogenic media produced similar phenotypes. Candidate matricellular proteins, clusterin and tenascin were identified by proteomics in the NCA group. CONCLUSIONS: NCs secreted important soluble factors capable of differentiating MSCs to a NP phenotype synthesizing high levels of proteoglycan while also resisting collagen fiber expression and hypertrophy, yet results were sensitive to the conditions in which media was generated (cells in alginate versus cells in their native tissue) so that further mechanistic studies optimizing culture conditions and defining important NC secreted factors are required. Matricellular proteins, such as clusterin and tenascin, are likely to be important to optimize differentiation of MSCs for maximum GAG production and reduced collagen fiber expression. PMID- 21627828 TI - Health status in the TORCH study of COPD: treatment efficacy and other determinants of change. AB - BACKGROUND: Little is known about factors that determine health status decline in clinical trials of COPD. OBJECTIVES: To examine health status changes over 3 years in the TORCH study of salmeterol+fluticasone propionate (SFC) vs. salmeterol alone, fluticasone propionate alone or placebo. METHODS: St George's Respiratory Questionnaire (SGRQ) was administered at baseline then every 6 months. MEASUREMENTS AND MAIN RESULTS: Data from 4951 patients in 28 countries were available. SFC produced significant improvements over placebo in all three SGRQ domains during the study: (Symptoms -3.6 [95% CI -4.8, -2.4], Activity -2.8 [95% CI -3.9, -1.6], Impacts -3.2 [95% CI -4.3, -2.1]) but the pattern of change over time differed between domains. SGRQ deteriorated faster in patients with Global Initiative for Chronic Obstructive Lung Disease (GOLD) stages III & IV relative to GOLD stage II (p < 0.001). There was no difference in the relationship between deterioration in SGRQ Total score and forced expiratory volume in one second (FEV1) decline (as % predicted) in men and women. Significantly faster deterioration in Total score relative to FEV1 % predicted was seen in older patients (>= 65 years) and there was an age-related change in Total score that was independent of change in FEV1. The relationship between deterioration in FEV1 and SGRQ did not differ in different world regions, but patients in Asia-Pacific showed a large improvement in score that was unrelated to FEV1 change. CONCLUSIONS: In addition to treatment effects, health status changes in clinical trials may be influenced by demographic and disease-related factors. Deterioration in health status appears to be fastest in older persons and those with severe airflow limitation. TRIAL REGISTRATION: ClinicalTrials.gov: NCT00268216. PMID- 21627829 TI - Refinement of primate copy number variation hotspots identifies candidate genomic regions evolving under positive selection. AB - BACKGROUND: Copy number variants (CNVs), defined as losses and gains of segments of genomic DNA, are a major source of genomic variation. RESULTS: In this study, we identified over 2,000 human CNVs that overlap with orthologous chimpanzee or orthologous macaque CNVs. Of these, 170 CNVs overlap with both chimpanzee and macaque CNVs, and these were collapsed into 34 hotspot regions of CNV formation. Many of these hotspot regions of CNV formation are functionally relevant, with a bias toward genes involved in immune function, some of which were previously shown to evolve under balancing selection in humans. The genes in these primate CNV formation hotspots have significant differential expression levels between species and show evidence for positive selection, indicating that they have evolved under species-specific, directional selection. CONCLUSIONS: These hotspots of primate CNV formation provide a novel perspective on divergence and selective pressures acting on these genomic regions. PMID- 21627830 TI - An industry perspective on Canadian patients' involvement in medical tourism: implications for public health. AB - BACKGROUND: The medical tourism industry, which assists patients with accessing non-emergency medical care abroad, has grown rapidly in recent years. A lack of reliable data about medical tourism makes it difficult to create policy, health system, and public health responses to address the associated risks and shortcomings, such as spread of infectious diseases, associated with this industry. This article addresses this knowledge gap by analyzing interviews conducted with Canadian medical tourism facilitators in order to understand Canadian patients' involvement in medical tourism and the implications of this involvement for public health. METHODS: Semi-structured phone interviews were conducted with 12 medical facilitators from 10 companies in 2010. An exhaustive recruitment strategy was used to identify interviewees. Questions focused on business dimensions, information exchange, medical tourists' decision-making, and facilitators' roles in medical tourism. Thematic analysis was undertaken following data collection. RESULTS: Facilitators helped their Canadian clients travel to 11 different countries. Estimates of the number of clients sent abroad annually varied due to demand factors. Facilitators commonly worked with medical tourists aged between 40 and 60 from a variety of socio-economic backgrounds who faced a number of potential barriers including affordability, fear of the unfamiliar, and lack of confidence. Medical tourists who chose not to use facilitators' services were thought to be interested in saving money or have cultural/familial connections to the destination country. Canadian doctors were commonly identified as barriers to securing clients. CONCLUSIONS: No effective Canadian public health response to medical tourism can treat medical tourists as a unified group with similar motivations for engaging in medical tourism and choosing similar mechanisms for doing so. This situation may be echoed in other countries with patients seeking care abroad. Therefore, a call for a comprehensive public health response to medical tourism and its effects should be coupled with a clear understanding that medical tourism is a highly diverse practice. This response must also acknowledge facilitators as important stakeholders in medical tourism. PMID- 21627831 TI - PhyDesign: an online application for profiling phylogenetic informativeness. AB - BACKGROUND: The rapid increase in number of sequenced genomes for species across of the tree of life is revealing a diverse suite of orthologous genes that could potentially be employed to inform molecular phylogenetic studies that encompass broader taxonomic sampling. Optimal usage of this diversity of loci requires user friendly tools to facilitate widespread cost-effective locus prioritization for phylogenetic sampling. The Townsend (2007) phylogenetic informativeness provides a unique empirical metric for guiding marker selection. However, no software or automated methodology to evaluate sequence alignments and estimate the phylogenetic informativeness metric has been available. RESULTS: Here, we present PhyDesign, a platform-independent online application that implements the Townsend (2007) phylogenetic informativeness analysis, providing a quantitative prediction of the utility of loci to solve specific phylogenetic questions. An easy-to-use interface facilitates uploading of alignments and ultrametric trees to calculate and depict profiles of informativeness over specified time ranges, and provides rankings of locus prioritization for epochs of interest. CONCLUSIONS: By providing these profiles, PhyDesign facilitates locus prioritization increasing the efficiency of sequencing for phylogenetic purposes compared to traditional studies with more laborious and low capacity screening methods, as well as increasing the accuracy of phylogenetic studies. Together with a manual and sample files, the application is freely accessible at http://phydesign.townsend.yale.edu. PMID- 21627832 TI - The effect of cathepsin K deficiency on airway development and TGF-beta1 degradation. AB - BACKGROUND: Cathepsin K, a cysteine protease predominantly expressed in osteoclasts, is a major drug target for the treatment of osteoporosis. Recent findings, however, indicate that cathepsin K is also involved in non-skeletal metabolism. The development of fibrotic phenotypes in lung and skin is a concern for cathepsin K inhibitors presently evaluated in clinical trials. Cathepsin K is expressed in lung tissue and has been implicated in lung fibrosis. However, little is known about the role of cathepsin K in airway development and its effect on TGF-beta1 degradation. METHODS: We investigated the effects of cathepsin K-deficiency on alterations in airway integrity, extracellular matrix composition, and TGF-beta1 expression and degradation. Lung homogenates of wild type and cathepsin K-deficient mice were used to evaluate their contents of collagen, glycosaminoglycans, and TGF-beta1. The accessibility of TGF-beta1 to cathepsin K-mediated degradation was determined in vitro and lung fibroblast proliferations in wild-type and cathepsin K-deficient cells were evaluated. RESULTS: Lung airway cathepsin K expression in wild-type mice remained constant between 1 and 6 months of age and the airway integrity was maintained. In contrast, after 2 months of age, all Ctsk-/- mice demonstrated increased airway epithelium thickness by 16-28%, a lower structural airway integrity (1-2 score units lower), elevated cytokeratin expression of 12%, increased alpha-actin and vimentin expression by 50% and 70%, increased area of smooth muscle cells by 15%, elevated hydroxyproline and GAGs content by 20% and 25%, and increased TGF-beta1 expression by 25%. TGF-beta1 proved an efficient substrate of cathepsin K and TGF beta1 protein content in lung was increased by a potent cathepsin inhibitor. Lung fibroblasts from Ctsk-/- mice after TGF-beta1 treatment showed increased proliferation rates, increased levels of TGF-beta1 by 30%, and increased ECM secretion. CONCLUSION: This study suggests that airway development is partly regulated by cathepsin K and that its expression contributes to the maintenance of the airway structural integrity. The anticipated use of therapeutic cathepsin K inhibitors needs to take potential changes in human lungs into consideration. PMID- 21627833 TI - "I did not intend to stop. I just could not stand cigarettes any more." A qualitative interview study of smoking cessation among the elderly. AB - BACKGROUND: Every year, more than 650,000 Europeans die because they smoke. Smoking is considered to be the single most preventable factor influencing health. General practitioners (GP) are encouraged to advise on smoking cessation at all suitable consultations. Unsolicited advice from GPs results in one of 40 60 smokers stopping smoking. Smoking cessation advice has traditionally been given on an individual basis. Our aim was to gain insights that may help general practitioners understand why people smoke, and why smokers stop and then remain quitting and, from this, to find fruitful approaches to the dialogue about stopping smoking. METHODS: Interviews with 18 elderly smokers and ex-smokers about their smoking and decisions to smoke or quit were analysed with qualitative content analysis across narratives. A narrative perspective was applied. RESULTS: Six stages in the smoking story emerged, from the start of smoking, where friends had a huge influence, until maintenance of the possible cessation. The informants were influenced by "all the others" at all stages. Spouses had vital influence in stopping, relapses and continued smoking. The majority of quitters had stopped by themselves without medication, and had kept the tobacco handy for 3-6 months. Often smoking cessation seemed to happen unplanned, though sometimes it was planned. With an increasingly negative social attitude towards smoking, the informants became more aware of the risks of smoking. CONCLUSION: "All the others" is a clue in the smoking story. For smoking cessation, it is essential to be aware of the influence of friends and family members, especially a spouse. People may stop smoking unplanned, even when motivation is not obvious. Information from the community and from doctors on the negative aspects of smoking should continue. Eliciting life-long smoking narratives may open up for a fruitful dialogue, as well as prompting reflection about smoking and adding to the motivation to stop. PMID- 21627834 TI - CTX-M-14 beta-lactamase-producing Citrobacter freundii isolated in Venezuela. AB - A clinical isolate of C. freundii with reduced susceptibility to extended spectrum beta-lactams from a woman with cystocele associated with recurrent urinary tract infection was analyzed. Susceptibility tests, double disk synergy tests (DDST) and enzymatic activity by the agar iodometric method suggested the presence of ESBLs. Conjugation experiments revealed the presence of a large conjugative plasmid (pLM07/20) with an exclusive FrepB replicon type (IncF/FIB). PCR analysis and sequencing confirmed the presence of the blaCTX-M-14 gene in the pLM07/20 from C. freundii.LM07/10. Although this is the first report of CTX-M-14 in Venezuela, we alert the medical community that future increase of these beta lactamases in our city could be due to dissemination of plasmids into bacterial populations. PMID- 21627835 TI - Static and dynamic mechanics of the murine lung after intratracheal bleomycin. AB - BACKGROUND: Despite its widespread use in pulmonary fibrosis research, the bleomycin mouse model has not been thoroughly validated from a pulmonary functional standpoint using new technologies. Purpose of this study was to systematically assess the functional alterations induced in murine lungs by fibrogenic agent bleomycin and to compare the forced oscillation technique with quasi-static pressure-volume curves in mice following bleomycin exposure. METHODS: Single intratracheal injections of saline (50 MUL) or bleomycin (2 mg/Kg in 50 MUL saline) were administered to C57BL/6 (n=40) and Balb/c (n=32) mice. Injury/fibrosis score, tissue volume density (TVD), collagen content, airway resistance (RN), tissue damping (G) and elastance coefficient (H), hysteresivity (eta), and area of pressure-volume curve (PV-A) were determined after 7 and 21 days (inflammation and fibrosis stage, respectively). Statistical hypothesis testing was performed using one-way ANOVA with LSD post hoc tests. RESULTS: Both C57BL/6 and Balb/c mice developed weight loss and lung inflammation after bleomycin. However, only C57BL/6 mice displayed cachexia and fibrosis, evidenced by increased fibrosis score, TVD, and collagen. At day 7, PV-A increased significantly and G and H non-significantly in bleomycin-exposed C57BL/6 mice compared to saline controls and further increase in all parameters was documented at day 21. G and H, but not PV-A, correlated well with the presence of fibrosis based on histology, TVD and collagen. In Balb/c mice, no change in collagen content, histology score, TVD, H and G was noted following bleomycin exposure, yet PV-A increased significantly compared to saline controls. CONCLUSIONS: Lung dysfunction in the bleomycin model is more pronounced during the fibrosis stage rather than the inflammation stage. Forced oscillation mechanics are accurate indicators of experimental bleomycin-induced lung fibrosis. Quasi-static PV curves may be more sensitive than forced oscillations at detecting inflammation and fibrosis. PMID- 21627836 TI - Baseline new bone formation does not predict bone loss in ankylosing spondylitis as assessed by quantitative computed tomography (QCT): 10-year follow-up. AB - BACKGROUND: To evaluate the relationship between bone loss and new bone formation in ankylosing spondylitis (AS) using 10-year X-ray, dual-energy x-ray absorptiometry (DXA) and quantitative computed tomography (QCT) follow-up. METHODS: Fifteen AS patients free from medical conditions and drugs affecting bone metabolism underwent X-ray, DXA and QCT in 1999 and 2009. RESULTS: In spine QCT a statistically significant (p = 0,001) decrease of trabecular bone mineral content (BMC) was observed (change +/- SD: 18.0 +/- 7.3 mg/cm3). In contrast, spine DXA revealed a significant increase of bone mineral density (change +/- SD: -0.15 +/- 0.14 g/cm2). The mean BMC, both at baseline and follow-up was significantly lower (p = 0.02 and p = 0.005, respectively) in advanced radiological group as compared to early radiological group. However, in multiple regression model after adjustment for baseline BMC, the baseline radiological scoring did not influence the progression of bone loss as assessed with QCT (p = 0.22, p for BMC*X-ray syndesmophyte scoring interaction = 0.65, p for ANOVA-based X-ray syndesmophyte scoring*time interaction = 0.39). Baseline BMC was the only significant determinant of 10-year BMC change, to date the longest QCT follow-up data in AS. CONCLUSIONS: In AS patients who were not using antiosteoporotic therapy spine trabecular bone density evaluated by QCT decreased over 10-year follow-up and was not related to baseline radiological severity of spine involvement. PMID- 21627837 TI - Mild hypothermia causes differential, time-dependent changes in cytokine expression and gliosis following endothelin-1-induced transient focal cerebral ischemia. AB - BACKGROUND: Stroke is an important cause of morbidity and mortality and few therapies exist thus far. Mild hypothermia (33 degrees C) is a promising neuroprotective strategy to improve outcome after ischemic stroke. However, its complete mechanism of action has not yet been fully elaborated. This study is the first to investigate whether this neuroprotection occurs through modulation of the neuroinflammatory response after stroke in a time-dependent manner. METHODS: The Endothelin-1 (Et-1) model was used to elicit a transient focal cerebral ischemia in male Wistar rats. In this model, the core and penumbra of the insult are represented by the striatum and the cortex respectively. We assessed the effects of 2 hours of hypothermia, started 20 minutes after Et-1 injection on neurological outcome and infarct volume. Furthermore, pro- and anti-inflammatory cytokine expression was determined using ELISA. Microgliosis and astrogliosis were investigated using CD-68 and GFAP staining respectively. All parameters were determined 8, 24, 72 hours and 1 week after the administration of Et-1. RESULTS: Et-1 infusion caused neurological deficit and a reproducible infarct size which increased up to 3 days after the insult. Both parameters were significantly reduced by hypothermia. The strongest reduction in infarct volume with hypothermia, at 3 days, corresponded with increased microglial activation. Reducing the brain temperature affected the stroke induced increase in interleukin-1beta and tumor necrosis factor alpha in the striatum, 8 hours after its induction, but not at later time points. Transforming growth factor beta increased as a function of time after the Et-1-induced insult and was not influenced by cooling. Hypothermia reduced astrogliosis at 1 and 3 days after stroke onset. CONCLUSIONS: The beneficial effects of hypothermia after stroke on infarct volume and functional outcome coincide with a time-dependent modulation of the cytokine expression and gliosis. PMID- 21627838 TI - Antifungal activity of redox-active benzaldehydes that target cellular antioxidation. AB - BACKGROUND: Disruption of cellular antioxidation systems should be an effective method for control of fungal pathogens. Such disruption can be achieved with redox-active compounds. Natural phenolic compounds can serve as potent redox cyclers that inhibit microbial growth through destabilization of cellular redox homeostasis and/or antioxidation systems. The aim of this study was to identify benzaldehydes that disrupt the fungal antioxidation system. These compounds could then function as chemosensitizing agents in concert with conventional drugs or fungicides to improve antifungal efficacy. METHODS: Benzaldehydes were tested as natural antifungal agents against strains of Aspergillus fumigatus, A. flavus, A. terreus and Penicillium expansum, fungi that are causative agents of human invasive aspergillosis and/or are mycotoxigenic. The yeast Saccharomyces cerevisiae was also used as a model system for identifying gene targets of benzaldehydes. The efficacy of screened compounds as effective chemosensitizers or as antifungal agents in formulations was tested with methods outlined by the Clinical Laboratory Standards Institute (CLSI). RESULTS: Several benzaldehydes are identified having potent antifungal activity. Structure-activity analysis reveals that antifungal activity increases by the presence of an ortho-hydroxyl group in the aromatic ring. Use of deletion mutants in the oxidative stress response pathway of S. cerevisiae (sod1Delta, sod2Delta, glr1Delta) and two mitogen-activated protein kinase (MAPK) mutants of A. fumigatus (sakADelta, mpkCDelta), indicates antifungal activity of the benzaldehydes is through disruption of cellular antioxidation. Certain benzaldehydes, in combination with phenylpyrroles, overcome tolerance of A. fumigatus MAPK mutants to this agent and/or increase sensitivity of fungal pathogens to mitochondrial respiration inhibitory agents. Synergistic chemosensitization greatly lowers minimum inhibitory (MIC) or fungicidal (MFC) concentrations. Effective inhibition of fungal growth can also be achieved using combinations of these benzaldehydes. CONCLUSIONS: Natural benzaldehydes targeting cellular antioxidation components of fungi, such as superoxide dismutases, glutathione reductase, etc., effectively inhibit fungal growth. They possess antifungal or chemosensitizing capacity to enhance efficacy of conventional antifungal agents. Chemosensitization can reduce costs, abate resistance, and alleviate negative side effects associated with current antifungal treatments. PMID- 21627839 TI - Phase II study of the combination carboplatin plus celecoxib in heavily pre treated recurrent ovarian cancer patients. AB - BACKGROUND: Cyclooxygenase-2 overexpression is associated with poor outcome and resistance to platinum-based chemotherapy in ovarian cancer. We evaluated the antitumor activity and safety of the combination carboplatin plus the COX-2 inhibitor celecoxib in recurrent heavily-treated OC patients. METHODS: Patients were administered oral celecoxib (400 mg/day) in combination with intravenous carboplatin (AUC5, q28). A Simon's two-stage design was employed. RESULTS: 45 patients were enrolled: 23 (51.1%) presented platinum-resistance, and 27 (60%) had received at least 3 prior regimens for recurrence. The response rate was 28.9% with 3 complete and 10 partial responses (median duration of response = 6 months). Only one (0.4%) G4 non-febrile neutropenia was observed; G3 neutropenia, anemia, or thrombocytopenia, were observed in 2.5%, 1.7%, and 1.7% of the cycles, respectively. G3-4 vomiting was reported in only 1.7%, and 0.4% of the cycles were associated with G3 dyspepsia or diarrhea or constipation. Only one patient experienced G3 hypertension associated to G2 hypersensitivity reaction. No differences in baseline versus post-treatment Quality of Life scores were observed. Median progression free survival and overall survival were 5 and 13 months, respectively. CONCLUSIONS: Celecoxib combined with carboplatin showed promising activity and it is well tolerated in heavily-treated recurrent ovarian cancer patients. TRIAL REGISTRATION NUMBER: NCT01124435 (ClinicalTrials.gov Identifier) and 935/03 (study ID numbers). PMID- 21627840 TI - Health-related productivity losses increase when the health condition is co morbid with psychological distress: findings from a large cross-sectional sample of working Australians. AB - BACKGROUND: The health condition of workers is known to impact on productivity outcomes. The relationship between health and productivity is of increasing interest amid the need to increase productivity to meet global financial challenges. Prevalence of psychological distress is also of growing concern in Australia with a two-fold increase in the prevalence of psychological distress in Australia from 1997-2005. METHODS: We used the cross-sectional data set from the Australian Work Outcomes Research Cost-benefit (WORC) study to explore the impacts of health conditions with and without co-morbid psychological distress, compared to those with neither condition, in a sample of approximately 78,000 working Australians. The World Health Organisation Health and Performance Questionnaire was used which provided data on demographic characteristics, health condition and working conditions. Data were analysed using negative binomial logistic regression and multinomial logistic regression models for absenteeism and presenteeism respectively. RESULTS: For both absenteeism and presenteeism productivity measures there was a greater risk of productivity loss associated when health conditions were co-morbid with psychological distress. For some conditions this risk was much greater for those with co-morbid psychological distress compared to those without. CONCLUSIONS: Co-morbid psychological distress demonstrates an increased risk of productivity loss for a range of health conditions. These findings highlight the need for further research to determine whether co-morbid psychological distress potentially exacerbates lost productivity. PMID- 21627841 TI - The nuclear envelope localization of DYT1 dystonia torsinA-DeltaE requires the SUN1 LINC complex component. AB - BACKGROUND: DYT1 dystonia is an autosomal dominant neurological condition caused by a mutation that removes a single glutamic acid residue (DeltaE) from the torsinA (torA) AAA+ protein. TorA appears to possess a nuclear envelope (NE) localized activity that requires Lamina-Associated-Polypeptide 1 (LAP1), which is an inner nuclear membrane localized torA-binding partner. Although hypoactive, the DYT1 dystonia torA-DeltaE isoform often concentrates in the NE, suggesting that torA-DeltaE also interacts with an NE-localized binding partner. RESULTS: We confirm that NE-localized torA-DeltaE does not co-immunoprecipitate with LAP1, and find that torA-DeltaE continues to concentrate in the NE of cells that lack LAP1. Instead, we find that variability in torA-DeltaE localization correlates with the presence of the SUN-domain and Nesprin proteins that assemble into the LINC complex. We also find that siRNA depletion of SUN1, but not other LINC complex components, removes torA-DeltaE from the NE. In contrast, the LAP1 dependent NE-accumulation of an ATP-locked torA mutant is unaffected by loss of LINC complex proteins. This SUN1 dependent torA-DeltaE localization requires the torA membrane association domain, as well as a putative substrate-interaction residue, Y147, neither of which are required for torA interaction with LAP1. We also find that mutation of these motifs, or depletion of SUN1, decreases the amount of torA-WT that colocalizes with NE markers, indicating that each also underlies a normal NE-localized torA binding interaction. CONCLUSIONS: These data suggest that the disease causing DeltaE mutation promotes an association between torA and SUN1 that is distinct to the interaction between LAP1 and ATP-bound torA. This evidence for two NE-localized binding partners suggests that torA may act on multiple substrates and/or possesses regulatory co-factor partners. In addition, finding that the DYT1 mutation causes abnormal association with SUN1 implicates LINC complex dysfunction in DYT1 dystonia pathogenesis, and suggests a gain-of-function activity contributes to this dominantly inherited disease. PMID- 21627842 TI - Histopathological grading of pediatric ependymoma: reproducibility and clinical relevance in European trial cohorts. AB - BACKGROUND: Histopathological grading of ependymoma has been controversial with respect to its reproducibility and clinical significance. In a 3-phase study, we reviewed the pathology of 229 intracranial ependymomas from European trial cohorts of infants (2 trials - SFOP/CNS9204) and older children (2 trials - AIEOP/CNS9904) to assess both diagnostic concordance among five neuropathologists and the prognostic utility of histopathological variables, particularly tumor grading. RESULTS: In phase 1, using WHO criteria and without first discussing any issue related to grading ependymomas, pathologists assessed and independently graded ependymomas from 3 of 4 trial cohorts. Diagnosis of grade II ependymoma was less frequent than grade III, a difference that increased when one cohort (CNS9204) was reassessed in phase 2, during which the pathologists discussed ependymoma grading, jointly reviewed all CNS9204 tumors, and defined a novel grading system based on the WHO classification. In phase 3, repeat independent review of two cohorts (SFOP/CNS9904) using the novel system was associated with a substantial increase in concordance on grading. Extent of tumor resection was significantly associated with progression-free survival (PFS) in SFOP and AIEOP, but not in CNS9204 and CNS9904. Strength of consensus on grade was significantly associated with PFS in only one trial cohort (AIEOP). Consensus on the scoring of individual histopathological features (necrosis, angiogenesis, cell density, and mitotic activity) correlated with PFS in AIEOP, but in no other trial. CONCLUSIONS: We conclude that concordance on grading ependymomas can be improved by using a more prescribed scheme based on the WHO classification. Unfortunately, this appears to have utility in limited clinical settings. PMID- 21627843 TI - Transcriptome map of plant mitochondria reveals islands of unexpected transcribed regions. AB - BACKGROUND: Plant mitochondria contain a relatively large amount of genetic information, suggesting that their functional regulation may not be as straightforward as that of metazoans. We used a genomic tiling array to draw a transcriptomic atlas of Oryza sativa japonica (rice) mitochondria, which was predicted to be approximately 490-kb long. RESULTS: Whereas statistical analysis verified the transcription of all previously known functional genes such as the ones related to oxidative phosphorylation, a similar extent of RNA expression was frequently observed in the inter-genic regions where none of the previously annotated genes are located. The newly identified open reading frames (ORFs) predicted in these transcribed inter-genic regions were generally not conserved among flowering plant species, suggesting that these ORFs did not play a role in mitochondrial principal functions. We also identified two partial fragments of retrotransposon sequences as being transcribed in rice mitochondria. CONCLUSION: The present study indicated the previously unexpected complexity of plant mitochondrial RNA metabolism. Our transcriptomic data (Oryza sativa Mitochondrial rna Expression Server: OsMES) is publicly accessible at [http://bioinf.mind.meiji.ac.jp/cgi-bin/gbrowse/OsMes/#search]. PMID- 21627844 TI - Economic impacts of illness in older workers: quantifying the impact of illness on income, tax revenue and government spending. AB - BACKGROUND: Long term illness has far reaching impacts on individuals, and also places a large burden upon government. This paper quantifies the indirect economic impacts of illness related early retirement on individuals and government in Australia in 2009. METHODS: The output data from a microsimulation model, Health&WealthMOD, was analysed. Health&WealthMOD is representative of the 45 to 64 year old Australian population in 2009. The average weekly total income, total government support payments, and total taxation revenue paid, for individuals who are employment full-time, employed part-time and not in the labour force due to ill health was quantified. RESULTS: It was found that persons out of the labour force due to illness had significantly lower incomes ($218 per week as opposed to $1167 per week for those employed full-time), received significantly higher transfer payments, and paid significantly less tax than those employed full-time or part-time. This results in an annual national loss of income of over $17 billion, an annual national increase of $1.5 billion in spending on government support payments, and an annual loss of $2.1 billion in taxation revenue. CONCLUSIONS: Illness related early retirement has significant economic impacts on both the individual and on governments as a result of lost income, lost taxation revenue and increased government support payments. This paper has quantified the extent of these impacts for Australia. PMID- 21627845 TI - Seeds for effective oligonucleotide design. AB - BACKGROUND: DNA oligonucleotides are a very useful tool in biology. The best algorithms for designing good DNA oligonucleotides are filtering out unsuitable regions using a seeding approach. Determining the quality of the seeds is crucial for the performance of these algorithms. RESULTS: We present a sound framework for evaluating the quality of seeds for oligonucleotide design. The F - score is used to measure the accuracy of each seed. A number of natural candidates are tested: contiguous (BLAST-like), spaced, transitions-constrained, and multiple spaced seeds. Multiple spaced seeds are the best, with more seeds providing better accuracy. Single spaced and transition seeds are very close whereas, as expected, contiguous seeds come last. Increased accuracy comes at the price of reduced efficiency. An exception is that single spaced and transitions constrained seeds are both more accurate and more efficient than contiguous ones. CONCLUSIONS: Our work confirms another application where multiple spaced seeds perform the best. It will be useful in improving the algorithms for oligonucleotide design. PMID- 21627846 TI - Variation in quality of preventive care for well adults in Indigenous community health centres in Australia. AB - BACKGROUND: Early onset and high prevalence of chronic disease among Indigenous Australians call for action on prevention. However, there is deficiency of information on the extent to which preventive services are delivered in Indigenous communities. This study examined the variation in quality of preventive care for well adults attending Indigenous community health centres in Australia. METHODS: During 2005-2009, clinical audits were conducted on a random sample (stratified by age and sex) of records of adults with no known chronic disease in 62 Indigenous community health centres in four Australian States/Territories (sample size 1839). MAIN OUTCOME MEASURES: i) adherence to delivery of guideline-scheduled services within the previous 24 months, including basic measurements, laboratory investigations, oral health checks, and brief intervention on lifestyle modification; and ii) follow-up of abnormal findings. RESULTS: Overall delivery of guideline-scheduled preventive services varied widely between health centres (range 5-74%). Documentation of abnormal blood pressure reading ([greater than or equal to]140/90 mmHg), proteinuria and abnormal blood glucose ([greater than or equal to]5.5 mmol/L) was found to range between 0 and > 90% at the health centre level. A similarly wide range was found between health centres for documented follow up check/test or management plan for people documented to have an abnormal clinical finding. Health centre level characteristics explained 13-47% of variation in documented preventive care, and the remaining variation was explained by client level characteristics. CONCLUSIONS: There is substantial room to improve preventive care for well adults in Indigenous primary care settings. Understanding of health centre and client level factors affecting variation in the care should assist clinicians, managers and policy makers to develop strategies to improve quality of preventive care in Indigenous communities. PMID- 21627847 TI - Are Australian immigrants at a risk of being physically inactive? AB - BACKGROUND: We examined whether physical activity risk differed between migrant sub-groups and the Australian-born population. METHODS: Data were drawn from the Australian National Health Survey (2001) and each resident's country of birth was classified into one of 13 regions. Data were gathered on each resident's physical activity level in the fortnight preceding the survey. Multivariable logistic regression, adjusted for potential confounders examined the risk of physical inactivity of participants from each of the 13 regions compared to the Australian born population. RESULTS: There was a greater prevalence of physical inactivity for female immigrants from most regions compared to male immigrants from a like region. Immigrants from South East Asia (OR 2.04% 95% CI 1.63, 2.56), Other Asia (OR 1.53 95% CI 1.10, 2.13), Other Oceania (1.81 95% CI 1.11, 2.95), the Middle East (OR 1.42 95% CI 0.97, 2.06 [note: border line significance]) and Southern & Eastern Europe are at a significantly higher risk of being physically inactive compared to those born in Australian. In contrast, immigrants from New Zealand (OR 0.77 95% CI 0.62, 0.94), the UK & Ireland (OR 0.82 95% CI 0.73, 0.92), and other Africa (OR 0.69 95% CI 0.51, 0.94) are at a significantly lower risk of being physically inactive compared to the Australian born population. CONCLUSION: Future research identifying potential barriers and facilitators to participation in physical activity will inform culturally sensitive physical activity programs that aim to encourage members of specific regional ethnic sub-groups to undertake physical activity. PMID- 21627848 TI - (-)-Epigallocatechin gallate attenuates NADPH-d/nNOS expression in motor neurons of rats following peripheral nerve injury. AB - BACKGROUND: Oxidative stress and large amounts of nitric oxide (NO) have been implicated in the pathophysiology of neuronal injury and neurodegenerative disease. Recent studies have shown that (-)-epigallocatechin gallate (EGCG), one of the green tea polyphenols, has potent antioxidant effects against free radical mediated lipid peroxidation in ischemia-induced neuronal damage. The purpose of this study was to examine whether EGCG would attenuate neuronal expression of NADPH-d/nNOS in the motor neurons of the lower brainstem following peripheral nerve crush. Thus, young adult rats were treated with EGCG (10, 25, or 50 mg/kg, i.p.) 30 min prior to crushing their hypoglossal and vagus nerves for 30 seconds (left side, at the cervical level). The treatment (pre-crush doses of EGCG) was continued from day 1 to day 6, and the animals were sacrificed on days 3, 7, 14 and 28. Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry and neuronal nitric oxide synthase (nNOS) immunohistochemistry were used to assess neuronal NADPH-d/nNOS expression in the hypoglossal nucleus and dorsal motor nucleus of the vagus. RESULTS: In rats treated with high dosages of EGCG (25 or 50 mg/kg), NADPH-d/nNOS reactivity and cell death of the motor neurons were significantly decreased. CONCLUSIONS: The present evidence indicated that EGCG can reduce NADPH-d/nNOS reactivity and thus may enhance motor neuron survival time following peripheral nerve injury. PMID- 21627849 TI - Detection and follow-up of chronic obstructive pulmonary disease (COPD) and risk factors in the Southern Cone of Latin America: the pulmonary risk in South America (PRISA) study. AB - BACKGROUND: The World Health Organization has estimated that by 2030, chronic obstructive pulmonary disease will be the third leading cause of death worldwide. Most knowledge of chronic obstructive pulmonary disease is based on studies performed in Europe or North America and little is known about the prevalence, patient characteristics and change in lung function over time in patients in developing countries, such as those of Latin America. This lack of knowledge is in sharp contrast to the high levels of tobacco consumption and exposure to biomass fuels exhibited in Latin America, both major risk factors for the development of chronic obstructive pulmonary disease. Studies have also demonstrated that most Latin American physicians frequently do not follow international chronic obstructive pulmonary disease diagnostic and treatment guidelines. The PRISA Study will expand the current knowledge regarding chronic obstructive pulmonary disease and risk factors in Argentina, Chile and Uruguay to inform policy makers and health professionals on the best policies and practices to address this condition. METHODS/DESIGN: PRISA is an observational, prospective cohort study with at least four years of follow-up. In the first year, PRISA has employed a randomized three-staged stratified cluster sampling strategy to identify 6,000 subjects from Marcos Paz and Bariloche, Argentina, Temuco, Chile, and Canelones, Uruguay. Information, such as comorbidities, socioeconomic status and tobacco and biomass exposure, will be collected and spirometry, anthropometric measurements, blood sampling and electrocardiogram will be performed. In year four, subjects will have repeat measurements taken. DISCUSSION: There is no longitudinal data on chronic obstructive pulmonary disease incidence and risk factors in the southern cone of Latin America, therefore this population-based prospective cohort study will fill knowledge gaps in the prevalence and incidence of chronic obstructive pulmonary disease, patient characteristics and changes in lung function over time as well as quality of life and health care resource utilization. Information gathered during the PRISA Study will inform public health interventions and prevention practices to reduce risk of COPD in the region. PMID- 21627850 TI - Major depressive disorder following terrorist attacks: a systematic review of prevalence, course and correlates. AB - BACKGROUND: Terrorist attacks are traumatic events that may result in a wide range of psychological disorders for people exposed. This review aimed to systematically assess the current evidence on major depressive disorder (MDD) after terrorist attacks. METHODS: A systematic review was performed. Studies included assessed the impact of human-made, intentional, terrorist attacks in direct victims and/or persons in general population and evaluated MDD based on diagnostic criteria. RESULTS: A total of 567 reports were identified, 11 of which were eligible for this review: 6 carried out with direct victims, 4 with persons in general population, and 1 with victims and general population. The reviewed literature suggests that the risk of MDD ranges between 20 and 30% in direct victims and between 4 and 10% in the general population in the first few months after terrorist attacks. Characteristics that tend to increase risk of MDD after a terrorist attack are female gender, having experienced more stressful situations before or after the attack, peritraumatic reactions during the attack, loss of psychosocial resources, and low social support. The course of MDD after terrorist attacks is less clear due to the scarcity of longitudinal studies. CONCLUSIONS: Methodological limitations in the literature of this field are considered and potentially important areas for future research such as the assessment of the course of MDD, the study of correlates of MDD or the comorbidity between MDD and other mental health problems are discussed. PMID- 21627851 TI - Feasibility and acceptability of point of care HIV testing in community outreach and GUM drop-in services in the North West of England: a programmatic evaluation. AB - BACKGROUND: In Liverpool, injecting drug users (IDUs), men-who-have-sex-with-men (MSM) and UK Africans experience a disproportionate burden of HIV, yet services do not reach out to these groups and late presentations continue. We set out to: increase testing uptake in targeted marginalized groups through a community and genitourinary medicine (GUM)-based point of care testing (POCT) programme; and conduct a process evaluation to examine service provider inputs and document service user perceptions of the programme. METHODS: Mixed quantitative, qualitative and process evaluation methods were used. Service providers were trained to use fourth generation rapid antibody/antigen HIV tests. Existing outreach services incorporated POCT into routine practice. Clients completed a semi-structured questionnaire and focus group discussions (FGDs) were held with service providers. RESULTS: Between September 2009 and June 2010, 953 individuals underwent POCT (GUM: 556 [59%]; community-based sites: 397 [42%]). Participants in the community were more likely to be male (p = 0.028), older (p < 0.001), of UK African origin (p < 0.001) and IDUs (p < 0.001) than participants from the GUM clinic. Seventeen new HIV diagnoses were confirmed (prevalence = 1.8%), 16 of whom were in risk exposure categories (prevalence: 16/517, 3.1%). Questionnaires and FGDs showed that clients and service providers were supportive of POCT, highlighting benefits of reaching out to marginalised communities and incorporating HIV prevention messages. CONCLUSIONS: Community and GUM clinic based POCT for HIV was feasible and acceptable to clients and service providers in a low prevalence setting. It successfully reached target groups, many of whom would not have otherwise tested. We recommend POCT be considered among strategies to increase the uptake of HIV testing among groups who are currently underserved. PMID- 21627852 TI - Hierarchical cluster-based partial least squares regression (HC-PLSR) is an efficient tool for metamodelling of nonlinear dynamic models. AB - BACKGROUND: Deterministic dynamic models of complex biological systems contain a large number of parameters and state variables, related through nonlinear differential equations with various types of feedback. A metamodel of such a dynamic model is a statistical approximation model that maps variation in parameters and initial conditions (inputs) to variation in features of the trajectories of the state variables (outputs) throughout the entire biologically relevant input space. A sufficiently accurate mapping can be exploited both instrumentally and epistemically. Multivariate regression methodology is a commonly used approach for emulating dynamic models. However, when the input output relations are highly nonlinear or non-monotone, a standard linear regression approach is prone to give suboptimal results. We therefore hypothesised that a more accurate mapping can be obtained by locally linear or locally polynomial regression. We present here a new method for local regression modelling, Hierarchical Cluster-based PLS regression (HC-PLSR), where fuzzy C means clustering is used to separate the data set into parts according to the structure of the response surface. We compare the metamodelling performance of HC PLSR with polynomial partial least squares regression (PLSR) and ordinary least squares (OLS) regression on various systems: six different gene regulatory network models with various types of feedback, a deterministic mathematical model of the mammalian circadian clock and a model of the mouse ventricular myocyte function. RESULTS: Our results indicate that multivariate regression is well suited for emulating dynamic models in systems biology. The hierarchical approach turned out to be superior to both polynomial PLSR and OLS regression in all three test cases. The advantage, in terms of explained variance and prediction accuracy, was largest in systems with highly nonlinear functional relationships and in systems with positive feedback loops. CONCLUSIONS: HC-PLSR is a promising approach for metamodelling in systems biology, especially for highly nonlinear or non-monotone parameter to phenotype maps. The algorithm can be flexibly adjusted to suit the complexity of the dynamic model behaviour, inviting automation in the metamodelling of complex systems. PMID- 21627853 TI - Prediction models for short children born small for gestational age (SGA) covering the total growth phase. Analyses based on data from KIGS (Pfizer International Growth Database). AB - BACKGROUND: Mathematical models can be developed to predict growth in short children treated with growth hormone (GH). These models can serve to optimize and individualize treatment in terms of height outcomes and costs. The aims of this study were to compile existing prediction models for short children born SGA (SGA), to develop new models and to validate the algorithms. METHODS: Existing models to predict height velocity (HV) for the first two and the fourth prepubertal years and during total pubertal growth (TPG) on GH were applied to SGA children from the KIGS (Pfizer International Growth Database)--1st year: N = 2340; 2nd year: N = 1358; 4th year: N = 182; TPG: N = 59. A new prediction model was developed for the 3rd prepubertal year based upon 317 children by means of the all-possible regression approach, using Mallow's C(p) criterion. RESULTS: The comparison between the observed and predicted height velocity showed no significant difference when the existing prediction models were applied to new cohorts. A model for predicting HV during the 3rd year explained 33% of the variability with an error SD of 1.0 cm/year. The predictors were (in order of importance): HV previous year; chronological age; weight SDS; mid-parent height SDS and GH dose. CONCLUSIONS: Models to predict growth to GH from prepubertal years to adult height are available for short children born SGA. The models utilize easily accessible predictors and are accurate. The overall explained variability in SGA is relatively low, due to the heterogeneity of the disorder. The models can be used to provide patients with a realistic expectation of treatment, and may help to identify compliance problems or other underlying causes of treatment failure. PMID- 21627854 TI - Microarrays, deep sequencing and the true measure of the transcriptome. AB - Microarrays first made the analysis of the transcriptome possible, and have produced much important information. Today, however, researchers are increasingly turning to direct high-throughput sequencing -- RNA-Seq -- which has considerable advantages for examining transcriptome fine structure -- for example in the detection of allele-specific expression and splice junctions. In this article, we discuss the relative merits of the two techniques, the inherent biases in each, and whether all of the vast body of array work needs to be revisited using the newer technology. We conclude that microarrays remain useful and accurate tools for measuring expression levels, and RNA-Seq complements and extends microarray measurements. PMID- 21627855 TI - No small feat: microRNA responses during vocal communication in songbirds. AB - Simply hearing the song produced by another bird of the same species triggers the regulation of microRNAs (miRNAs) in high-order auditory parts of the zebra finch brain. Some of the identified miRNAs appear to be unique to birds, possibly to songbirds. These findings, reported in BMC Genomics, highlight the complexities of gene regulation associated with vocal communication and point to possible key regulators of song-triggered gene networks. PMID- 21627856 TI - DNA replication: archaeal oriGINS. AB - GINS is an essential eukaryotic DNA replication factor that is found in a simplified form in Archaea. A new study in this issue of BMC Biology reveals the first structure of the archaeal GINS complex. The structure reveals the anticipated similarity to the previously determined eukaryotic complex but also has some intriguing differences in the relative disposition of subunit domains. PMID- 21627857 TI - A proposal: a comprehensive platform to characterize tumors in Chinese and improve success in cancer drug discovery and development. AB - Cancer is a collection of complex diseases in which cell proliferation and apoptosis are dysregulated due to the acquisition of genetic changes in cancer cells. These genetic changes, combined with the interrelated physiologic adaptations of neo-angiogenesis, recruitment of stromal support tissues, and suppression of immune recognition, are measurable characteristics in tumor gene expression profiles and biochemical pathways. These measures can lead to identification of disease drivers and, ultimately, can be used to assign therapy. With advances in RNA sequencing technologies, the ability to simultaneously measure all genetic and gene expression changes with a single technology is now possible. The ability to create a comprehensive catalog of genotypic and phenotypic changes in a collection of histologically similar but otherwise distinct tumors should allow for a more precise positioning of existing targeted therapies and identification of new targets for intervention. PMID- 21627858 TI - MicroRNA therapeutics: principles, expectations, and challenges. AB - MicroRNAs (miRNAs) are a class of highly abundant non-coding RNA molecules that are involved in several biological processes. Many miRNAs are often deregulated in several diseases including cancer. There is substantial interest in exploiting miRNAs for therapeutic applications. In this editorial, we briefly review current advances in the use of miRNAs or antisense oligonucleotides (antagomirs) for such therapies. One of the key issues related to therapy using miRNAs is degradation of naked particles in vivo. To overcome this limitation, delivery systems for miRNA-based therapeutic agents have been developed, which hold tremendous potential for improving therapeutic outcome of cancer patients. PMID- 21627859 TI - Apoptosis and the target genes of microRNA-21. AB - MicroRNA-21 (miR-21) is frequently up-regulated in cancer and the majority of its reported targets are tumor suppressors. Through functional suppression, miR-21 is implicated in practically every walk of oncogenic life: the promotion of cell proliferation, invasion and metastasis, genome instability and mutation, inflammation, replicative immortalization, abnormal metabolism, angiogenesis, and evading apoptosis, immune destruction, and growth suppressors. In particular, miR 21 is strongly involved in apoptosis. In this article, we reviewed the experimentally validated targets of miR-21 and found that two thirds are linked to intrinsic and/or extrinsic pathways of cellular apoptosis. This suggests that miR-21 is an oncogene which plays a key role in resisting programmed cell death in cancer cells and that targeting apoptosis is a viable therapeutic option against cancers expressing miR-21. PMID- 21627861 TI - Research progresses in the pathogenesis of anaplastic large cell lymphoma. AB - Anaplastic large cell lymphoma (ALCL) is a distinct subset of T-cell non Hodgkin's lymphoma. As a consequence of its low incidence, general pathogenic consideration of ALCL is lacking. In this review, we summarize the pathogenesis, epidemiology, clinical manifestations, and treatment of ALCL, so as to better understand key stages of the development of this disease and provide valuable information for future treatment. PMID- 21627860 TI - Single-nucleotide polymorphisms among microRNA: big effects on cancer. AB - MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression at the transcriptional or posttranscriptional level. Many miRNAs are found to play a significant role in cancer development either as tumor suppressor genes or as oncogenes. Examination of tumor-specific miRNA expression profiles in diverse cancers has revealed widespread deregulation of these molecules, whose loss and overexpression respectively have diagnostic and prognostic significance. Genetic variations, mostly single-nucleotide polymorphisms (SNPs) within miRNA sequences or their target sites, have been found to be associated with many kinds of cancers. In this review, we summarize the current knowledge of miRNAs including their biogenesis and role in cancer development, and finally, how SNPs among miRNAs affect miRNA biogenesis and contribute to cancer. PMID- 21627862 TI - RNA interference of pax2 inhibits growth of transplanted human endometrial cancer cells in nude mice. AB - The development of human endometrial carcinoma (HEC) is a complex pathologic process involves several oncogenes and tumor suppressor genes. The full-length paired-box gene 2 (pax2), a recently discovered oncogene, promotes cell proliferation and growth and inhibits apoptosis of HEC cells. Here, we examined the effect of pax2 small interfering RNA (siRNA) on the growth of transplanted HEC cells in nude mice. The expression of Pax2 in 21 cases of normal endometrium and 38 cases of HEC was examined by immohistochemistry (IHC). HEC models were developed by subcutaneously transferring HEC cells into nude mice, followed by treatment with empty lentivirus vector, lentivirus vector-based pax2 siRNA, and phosphate buffered saline, respectively. Four weeks later, tumor size was measured, tumor inhibition rate was calculated, and histological analyses were conducted after staining with hematoxylin and eosin. The expression of Pax2 and Bcl-2 was detected by Western blot; proliferating cell nuclear antigen (PCNA) was detected by IHC. Significant differences were observed in the positive rate of Pax2 between normal endometrium and HEC (14.2% vs. 60.5%, P < 0.01). The expression index of Pax2 in well differentiated tumors was 1.88 +/- 1.68, much lower than that in tumors of moderate (3.07 +/- 1.96, P < 0.05) or poor differentiation (5.45 +/- 2.76, P <0.01). Tumor necrosis increased, nuclear basophilia stain decreased, tumor growth was inhibited, and PCNA, Pax2, and Bcl-2 expression was reduced in HEC models treated with pax2 siRNA. These results indicate that Pax2 expression is related to HEC tumor biology with the increased expression of Pax2 correlated to malignancy. pax2 siRNA down-regulates Pax2 expression and inhibits tumorigenesis of HEC in nude mice, possibly due to cell apoptosis and the inhibition of tumor proliferation induced by down-regulation of Bcl-2. PMID- 21627863 TI - Identification of plasma microRNA-21 as a biomarker for early detection and chemosensitivity of non-small cell lung cancer. AB - Studies have shown cell-free microRNA (miRNA) circulating in the serum and plasma with specific expression in cancer, indicating the potential of using miRNAs as biomarkers for cancer diagnosis and therapy. This study was to investigate whether plasma miRNA-21 (miR-21) can be used as a biomarker for the early detection of non-small cell lung cancer (NSCLC) and to explore its association with clinicopathologic features and sensitivity to platinum-based chemotherapy. We used real-time RT-PCR to investigate the expression of miR-21 in the plasma of 63 NSCLC patients and 30 healthy controls and correlated the findings with early diagnosis, pathologic parameters, and treatment. Thirty-five patients (stages IIIB and IV) were evaluable for chemotherapeutic responses: 11 had partial response (PR); 24 had stable and progressive disease (SD+ PD). Plasma miR-21 was significantly higher in NSCLC patients than in age- and sex-matched controls (P < 0.001). miR-21 was related to TNM stage (P < 0.001), but not related to age, sex, smoking status, histological classification, lymph node status, and metastasis (all P > 0.05). This marker yielded a receiver operating characteristic (ROC) curve area of 0.775 (95% CI: 0.681- 0.868) with 76.2% sensitivity and 70.0% specificity. Importantly, miR-21 plasma levels in PR samples were several folds lower than that in SD plus PD samples (P = 0.049), and were close to that in healthy controls (P = 0.130). Plasma miR-21 can serve as a circulating tumor biomarker for the early diagnosis of NSCLC and is related to the sensitivity to platinum-base chemotherapy. PMID- 21627864 TI - Loss of A-type lamin expression compromises nuclear envelope integrity in breast cancer. AB - Through advances in technology, the genetic basis of cancer has been investigated at the genomic level, and many fundamental questions have begun to be addressed. Among several key unresolved questions in cancer biology, the molecular basis for the link between nuclear deformation and malignancy has not been determined. Another hallmark of human cancer is aneuploidy; however, the causes and consequences of aneuploidy are unanswered and are hotly contested topics. We found that nuclear lamina proteins lamin A/C are absent in a significant fraction (38%) of human breast cancer tissues. Even in lamin A/C-positive breast cancer, lamin A/C expression is heterogeneous or aberrant (such as non-nuclear distribution) in the population of tumor cells, as determined by immunohistology and immunofluorescence microscopy. In most breast cancer cell lines, a significant fraction of the lamin A/C-negative population was observed. To determine the consequences of the loss of lamin A/C, we suppressed their expression by shRNA in non-cancerous primary breast epithelial cells. Down regulation of lamin A/C in breast epithelial cells led to morphological deformation, resembling that of cancer cells, as observed by immunofluorescence microscopy. The lamin A/C-suppressed breast epithelial cells developed aneuploidy as determined by both flow cytometry and fluorescence in situ hybridization. We conclude that the loss of nuclear envelope structural proteins lamin A/C in breast cancer underlies the two hallmarks of cancer aberrations in nuclear morphology and aneuploidy. PMID- 21627865 TI - Enrichment of osteosarcoma stem cells by chemotherapy. AB - Osteosarcoma is the most common primary malignant bone cancer in children and adolescents. Emerging evidence has suggested that the capability of a tumor to grow is driven by a small subset of cells within a tumor, termed cancer stem cells (CSCs). Although several methods have been explored to identify or enrich CSCs in osteosarcoma, these methods sometimes seem impractical, and chemotherapy enrichment for CSCs in osteosarcoma is rarely investigated. In the present study, we found that short exposure to chemotherapy could change the morphology of osteosarcoma cells and increase sarcosphere formation in vitro, as well as increase tumor formation in vivo. Furthermore, methotrexate (MTX)-resistant U2OS/MTX300 osteosarcoma cells were larger in size and grew much more tightly than parental U2OS cells. More importantly, U2OS/MTX300 cells possessed a higher potential to generate sarcospheres in serum-free conditions compared to parental U2OS cells. Also, U2OS/MTX300 cells exhibited the side population (SP) phenotype and expressed CSC surface markers CD117 and Stro-1. Notably, U2OS/MTX300 cells showed a substantially higher tumorigenicity in nude mice relative to U2OS cells. Therefore, we conclude that chemotherapy enrichment is a feasible and practical way to enrich osteosarcoma stem cells. PMID- 21627866 TI - Giant cell arteritis presenting with bilateral loss of vision and jaw pain: reminder of a potentially devastating condition. AB - A case of giant cell arteritis was diagnosed in a woman who presented with bilateral loss of vision that did not improve with intravenous corticosteroid therapy. Preceding her vision loss, the patient had other symptoms consistent with this diagnosis, notably significant jaw pain and arthralgias. The diverse symptoms of giant cell arteritis are discussed, along with the features that can be used to distinguish jaw pain associated with this condition from the pain of temporomandibular joint pathology. An increased awareness of giant cell arteritis should lead to earlier diagnosis and treatment and avoidance of the devastating consequences of this condition. PMID- 21627867 TI - Utilization of dental services by children in low-income families in Alberta. AB - OBJECTIVE: To explore the utilization of dental services for children among low income families receiving assistance from 2 provincial health benefit programs in Alberta. METHODS: A survey questionnaire was used in telephone interviews with 820 randomly selected clients of the Alberta Child Health Benefit (ACHB) and the Alberta Adult Health Benefit (AAHB) programs. Data related to utilization of dental services were analyzed. RESULTS: Among respondents to the questionnaires, 377 (93.1%) of 405 ACHB clients and 356 (85.8%) of 415 AAHB clients agreed that the programs helped them or their children to obtain dental services that they would not otherwise be able to receive. However, only 222 (54.8%) of the 405 ACHB respondents and 136 (57.4%) of 237 respondents with children covered by the AAHB program reported that their youngest child had received at least 1 dental service in the 12 months before the survey. Children in the 2 youngest age cohorts (i.e., those 4 years of age or younger) were less likely to have received each of several specific dental services, including a dental examination or checkup, and children 5 to 14 years of age were more likely to have received these dental services. The most used dental service for all age groups was a dental examination or checkup, and the least used was extraction. CONCLUSIONS: Despite the great need, low-income families underutilized the dental benefits for children offered by Alberta Employment and Immigration, which are designed to assist low-income Albertans. Parental awareness about public funding for dental services that is available did not seem to provide enough motivation to seek dental care for young children. PMID- 21627868 TI - A large expansion in the mandible: clinical features and differential diagnosis. PMID- 21627869 TI - Effects of external bleaching on restorative materials: a review. AB - With the increasingly commercial emphasis on dental esthetics, patients have become more interested in improving the appearance of their smile. For many, whiter teeth is their chief goal. However, when considering dental bleaching, practitioners need to take certain precautions. Over the past decade, multiple studies have evaluated the effects of bleaching agents on restorative materials. This article reviews their conclusions, focusing mainly on the clinical impact these agents can have on amalgam, porcelain, ormocer, glass ionomer, compomer and composite resin restorations. PMID- 21627870 TI - Behaviour management of a pediatric dental patient: audiovisual presentation. PMID- 21627871 TI - Management of a broken needle in the pterygomandibular space following a Vazirani Akinosi block: case report. AB - Although needle breakage is a rare complication of local anesthetic administration in the oral cavity, it can result in a high degree of morbidity and legal action against the practitioner. This is a case report of a 48-year-old woman who was referred for removal of a broken needle following a Vazirani Akinosi nerve block. Management involved removal of the needle in a surgical setting with fluoroscopic guidance and 2-point reference. PMID- 21627872 TI - [Validation of a questionnaire to evaluate alcohol related behaviours in excessive drinkers]. AB - The excessive consumption of alcohol and alcoholic drinks is an habit with high socioeconomic costs and a strong impact on health that requires research aiming at establishing adequate political options concerning trends, observed behaviours and consumption. OBJECTIVES: To validate an instrument to evaluate the consumption of the different kinds of alcoholic drinks and alcohol-related behaviours; to assess the determinants of alcoholic drinks consumption. TYPE OF STUDY: Validation study of an instrument to assess alcohol-related behaviours. PLACE: Coimbra. POPULATION: Excessive alcohol consumers. METHODS: Cross-sectional study based on a questionnaire and biochemical markers assessment, aiming to validate an instrument to evaluate alcohol consumption and alcohol-related behaviours. RESULTS: A correlation between age at first contact with alcohol and the beginning of the regular consumption(r = 0,72; p < 0,001) was found. Alcohol consumption in the last 12 months correlated with - % CDT (r = 0,54; p < 0,001) and with gamma-GT (r = 0,47; p < 0,001). CONCLUSIONS: The validity of the instrument developed to evaluate alcohol-related behaviours is acceptable and therefore it may be used to establish hierarchy between levels of alcohol consumption defined by categories of ingestion and negative consequences. PMID- 21627873 TI - [Characteristics of alcoholics associated to longer adherence to an out-treatment program]. AB - INTRODUCTION: Low adherence rates are frequently found in alcoholics' treatment programs. Many characteristics of alcoholics have already been associated to longer adherence or to higher drop-out rates. Adherence is an outcome already pointed as a simple and practical measure to evaluate treatment results. This study aimed to identify characteristics of alcoholics of a public outpatient treatment program associated to superior adherence, i.e., to longer permanence periods in treatment. METHOD: This study included 634 alcoholics; from these, 329 concluded the program's assessment stage and 305 did not conclude or fulfill the program's exclusion criteria. Initially, we analyzed socio-demographic differences between patients who completed or did not complete assessment. Crosstabs were performed to verify the association of all other 814 variables in the data bank to adherence to treatment. Logistic regression of variables found at least marginally significant was the performed: first within each dimension involved, contributing to the selection of variables for the multidimensional model. Pearson's Qui-square test was used to verify possible associations. RESULTS: Only two variables significantly differed patients who completed and did not complete assessment: place of birth and mean age. In bivariate analysis, 65 variables were associated to adherence; however, only five remained significant (p < 0.05) after logistic regression. In the final model, variables significantly associated to adherence were: did not eat while drinking; evaluated as having high insight level; (presented DSM-IV criterion 6 for Dependence (important social, occupational, or recreational activities are given up or reduced because of substance use); when increased consumption, drank to relieve chronic malaise; abdominal pain in the last 30 days before evaluation. CONCLUSION: The results of this study indicate that, differently from preliminary conclusions, the therapeutic standard used in the program evaluated--namely its focus on the approach of psychiatric comorbidity--did not remain as one of the most relevant factors associated to longer adherence to treatment. Therefore, the results support the need of new studies directed to the assessment of these subgroups that more quickly dropped-out of the treatment routinely offered, in order that diagnose and treatment approaches may be altered in order to become more efficient and effective to its clients. PMID- 21627874 TI - [After the booze comes the hangover: a perspective of alcohol consumption in young]. AB - Until today, little relevance has been given to the main cause of alcohol consumption related morbidity in young population, the so called "the next day morning alcohol hangover". The hangover is defined by the presence of symptoms connected to excessive alcohol consumption and its total metabolism, with severity enough to disturb responsibilities and daily life activities. Numerous observations show us that generally the young populations tend to be involved in a series of behaviors to cope with the unpleasant effects of a night of immoderate alcohol consumption. Through an empirical evaluation, it will be argued in this study the circumstances implicated in alcohol hangover and which behaviors the young population normally tends to be involved in order to attenuate it. The sample comprised 134 university students (1o year). It can be concluded that the frequency of the behaviors to cope with alcohol hangover translates the need to remove or alleviate in an accurate and symptomatic way the most reiterated effects of aversive alcohol hangover cluster. This work provides reliable information that could be employed from an educational point of view, while we profound which cognitive, behavior and physiological mechanisms occurs during an episode of alcohol hangover. Taken into account that the consumption of alcoholic beverage is a normative behavior in adolescence, we propose a realistic perspective of the phenomenon (more than ideological and utopic), that encompass the maximum delay of the beginning of alcohol consumption in young, educating them in what concerns the potential harm of its consumption, incorporated in a broad perspective of promotion of an healthy life style and of proximity with the adolescent. PMID- 21627875 TI - [Medicines, anxiety and depression]. AB - OBJECTIVE: To ascertain the opinion about anxiety, depression and the medicines to treat them by gender, labour activity and age. MATERIAL AND METHODS: validated questionnaire; patients attending a Central Portugal Health Centre; doctors and nurses that proposed the questionnaire to all attending patients. Cross sectional observational study, by anonymous and secret self-fulfilment, in a convenience sample drawn from a population composed by all those attending the Health Centre between the 22nd June and the 3rd July (both inclusive) 2009. Descriptive and inferential statistics were performed by means of a SPSS 11.0 statistical package. Age groups were defined as: < 36 years, 36-55 years, =56 years and professional activity groups as non-actives (students, retired and unemployed) and actives (housewives, and workers in primary, secondary and tertiary sectors). RESULTS: A n = 281 sample was studied age distribution of < 36 years = 47,7%, 36 55 years = 36,7% and = 56 years = 15,6%, mostly active (69,4% actives) and predominantly female (71,6% women). Age distribution is normal (Kolmogorov Smirnov Z = 1,644 2 tailed p = 0,009). As results of agreement vs disagreement, showing the significant differences, 'In general medicines for anxiety and for depression are capable of making me feel well' (71,4% vs 15,2%, p = 0,035 by age group), 'In general medicines can relieve the unpleasant sensations caused by day by day stress of actual society' (67,7% vs 22,6%, ns), 'If necessary medicines can relieve my anxiety or depression problems' (52,0% vs 34,1%, p = 0,000 by gender) and 'Anxiety and depression problems are displays of body illnesses' (42,8% vs 33,2%, ns). There is a majority of disagreement for 'In general medicines can, by themselves cure depression or anxiety' (72,5% vs 20,7%, p = 0,013 by sex), 'In general medicines for anxiety or depression can help change the way one sees the problems' (49,4% vs 43,4%, p = 0,041 by age group, p = 0,004 by sex, p = 0,002 by group of professional activity), 'I can feel good just by taking medicines' (71,4% vs 19,8%, p =0,008 by sex, p = 0,006 by professional activity group) and 'Medicines can bring more joy to my life' (58,5% vs 23,9%, p = 0,008 by sex, p = 0,006 by professional activity group). Anxiety problems and depression problems are demonstrations of body diseases, 42,8% vs 33,2%, ns). CONCLUSIONS: The pharmacological treatment for anxiety or depression must be individualised by sex, gender, age and activity of patients. In general medicines are judged capable of curing anxiety and depression problems but in the particular own case their activity is considered. Anxiety problems and depression problems are considered as manifestations of the body's disease. PMID- 21627876 TI - [Mental retardation: case series of Dona Estefania Hospital child development centre]. AB - Mental retardation (MR) consists of a sub average general intellectual functioning (IQ < 70) that is associated with significant limitations in adaptative functioning which begins before age 18 years (DSM IV). It's our intention to characterize the children's population with MR observed at the Dona Estefania Hospital Development Center, trough a retrospective study of the children which were assessed between January 2005 and June 2007. Epidemiologic data, like the severity of DM, organic aetiologies, co-morbidity and therapeutical and educational intervention, were the parameters evaluated. There were 232 children assessed and 185 had MR, mainly boys (58%): 112 (46%) had mild MR, 54 (29%) had moderate MR, 17 (9%) had severe DM and 2 (1%) profound MR. Aetiologies were defined on 86 children (46%); the highest rate of diagnosis occurred on the most severe cases of MR. The aetiologies most frequently found were the genetic diseases, prematurity and associated comorbidity, with high variety of aetiologies. Co-morbidity was detected on 123 children (66%), being the most frequent the ophthalmologic (57 children, 46%). The totality of children with MR were proposed for early intervention (47%) and special education (58%). The data obtained from the clinical and functional characterization of the children accompanied by the Development Center of Dona Estefania Hospital with MR diagnosis was similar to the data described in literature. However, some data differ from other case series due, among other things, to etiological group differences, different age groups and different assistential politics. PMID- 21627877 TI - [Transsexuality: experiencing the transition process in the context of health care services]. AB - Genital reconstruction surgery and hormonal treatment for transsexuals are performed in health care services following a long and complex evaluation process generally based on the guidelines proposed by the World Professional Association for Transgender Health. Though important to ensure these individuals' health and well-being, such procedures may conflict with their reality, raising obstacles not only for the candidates but also for professionals who deal with them. The reality of this population has been the focus of recent studies, whose results are to be considered in the new version (the seventh) of the procedures, currently under elaboration. In Portugal, the experience transsexuals have of their transition process, and of that process within health care services is still unknown. Yet, integrating that experience with these guidelines may contribute to better adapting procedures to their target population, decreasing obstacles facing the parties involved and improving available services. Seeking to bridge this gap and to contribute to the increased integration of population's needs and professionals' procedures, the current work examines transsexuals' experiences of their transition process, highlighting aspects that constitute difficulties to it or else facilitate it. The study is based on in-depth interviews with biological male transsexuals who request hormonal and surgical interventions to acquire a female physiology. Data were content-analyzed in the software program NUD*IST. The analysis reveals several themes associated with positive experiences in the sample's transition process (e.g., presence and accessibility of services dealing with this issue, presence of guidelines, setting the surgery date), negative experiences (e.g., process duration and uncertainties, professional treatment received), and experiences which cut across both (e.g., quality and variety of available services), as well as some paradoxes. In general, the interviewees defend the existence of non-rigid guidelines used in transgender evaluation, as well as flexibility in the treatment offered by professionals, based on knowledge and respect for each case and on updated information on this domain. Some procedures that may contribute to a more effective handling of the transition process are outlined based on identified themes. The articulation of diverse contexts and services (medical, legal, professional) is necessary for the improved success of the transition. PMID- 21627878 TI - [Portuguese version of the Markova and Berrios Insight Scale]. AB - Lack of insight into mental disorder is a frequent phenomenon in patients affected by a psychotic disorder and can be an important predicting factor involving the course of the disease mainly regarding compliance with treatment. The aim of the present study was to validate the Portuguese version of the Markova and Berrios Insight Scale, a self-report instrument that allow us to evaluate insight into psychotic illness. After a translation-back translation process, the Portuguese version of the scale was administered to a sample of 83 psychotic patients (DSM-IV). The results demonstrated that the Markova and Berrios Insight Scale achieved a excellent viability, internal reliability (a = 0,80). These coefficients were similar to those found by the researchers who developed this scale. The Markova and Berrios Insight Scale has proven to be easily applied and deemed an useful and valid instrument in researches involving psychotic subjects. PMID- 21627879 TI - [Quality of life in oncology: electronic device to collect data]. AB - INTRODUCTION: We aimed to evaluate an automated method for collecting data for cancer patients' quality of life (QoL). We compared the traditional method of collecting data on paper with the new collecting method using a touch screen computer. We also studied its reliability as a feasible alternative to evaluate the QoL of cancer patients in real time. METHODS: 200 cancer patients completed the EORTC QLQ-C30 on paper and on the computer with a time interval of one hour. The first 100 patients answered first on paper and, subsequently, on the computer; the remaining patients followed the opposite procedure. RESULTS: There were no statistically significant differences between the paper and the computer answers. We observed high correlations and large agreements between them. The best correlation values were obtained for the subscales diarrhea and physical functioning. Besides these findings, we observed a higher acceptability among patients using the computer when comparing with the use of paper. CONCLUSION: The computer version of the QLQ-C30 showed similar results compared to the paper version and it proved to be better accepted and tolerated by the cancer patients included in the study. This method can be valid in oncology setting and useful for monitoring the patients' QoL in daily practice, in real time, and it could be a useful means to support clinical decision making. PMID- 21627880 TI - [Economic analysis of rituximab in combination with cyclophosphamide, vincristine and prednisolone in the treatment of patients with advanced follicular lymphoma in Portugal]. AB - OBJECTIVE: Evaluate costs and benefits of rituximab in combination with cyclophosphamide/vincristine/prednisolone chemotherapy regimen (R-CVP), in previously untreated patients with indolent non-Hodgkin lymphoma (NHL), compared to CVP alone from a Portuguese National Health System (NHS) perspective. METHODS: Cost-effectiveness (Life Years Gained--LYG) and cost-utility analysis (Quality Adjusted Life Years--QALYs) were performed for a time horizon of 10 years, according to a Markov economic model with three health states (progression free survival, progression and death) and monthly cycles for a population of previously untreated patients with indolent NHL. Data from a phase III clinical trial was used and expanded to include unpublished 53-month median follow-up data. Survival after first-line therapy was estimated from the Scotland and Newcastle Lymphoma Group registry data and utilities were derived from a study in the UK performed in patients with follicular lymphoma. Resource consumption was estimated by a Portuguese expert panel (Delbecq Panel). Costs were calculated from the Portuguese NHS perspective through official data with prices updated to 2008. Only direct medical costs were considered. Costs and clinical outcomes were discounted at 5% per annum. Deterministic and probabilistic sensitivity analysis were performed around assumptions on the time horizon, costs, utilities and excess mortality rate due to progression applied in the base-case analysis. RESULTS: The 10-year base-case analysis showed a lower total cost per patient with CVP alone (? 85,838) in comparison with R-CVP (? 87,774). Life expectancy and Quality adjusted life expectancy per patient were higher with R-CVP (6.361 and 4.166, respectively) than with CVP alone (5.557 and 3.438, respectively), representing increases of 0.804 in LYG and 0.728 (8.7 months) in QALYs gained. The incremental cost per LYG was ? 2,407 and the incremental cost per QALY gained was ? 2,661. The probabilistic sensitivity analysis confirmed the robustness of the base-case analysis results. CONCLUSIONS: This study demonstrates that the combination R-CVP in previously untreated indolent NHL patients improves life expectancy and is a cost-effective alternative to CVP in Portugal. PMID- 21627881 TI - [Patient centered practice in internal medicine]. AB - We made a cross-sectional study aimed at 50 professionals (18 doctors, 32 nurses) of a tertiary hospital Internal Medicine (IM) ward, focusing the relevant knowledge in various areas of Patient-Centered Care in Chronic Disease: symptom control, pharmacology and palliative prognostic discussion. Almost 98% believe that most patients need strategies for symptomatic care, which died in hospital in considerable suffering (68%). Provision of palliative care in the community was rarely established with the primary health team. 90% were favourable on the creation of a hospital palliative care team. Around 57% find essential to prognosticate before thinking about mitigation strategies. While 75% of professionals had already discussed end-of-life directives with, at least, one patient, only one case could be formalized in writing. The rate of use of scales for assessing the intensity of pain was less than 50% and 38% did not indicate major opioids for the treatment of moderate intensity pain. These were considered contra-indicated for relief of dyspnoea in chronic obstructive pulmonary disease by 20% of professionals and 55% of those were unaware that its use and titration is governed by the same principles used in pain control. Around 44% of the respondents had already used the subcutaneous route for administration of drugs and 58% for hydration. Despite the team recognition of the potential for suffering of patients and the need for mitigation strategies, they remain linked to prognosis and not to symptomatic complexity. There are gaps in regard to control of pain, dyspnoea and in continuity of care. PMID- 21627882 TI - [Cognition, social cognition and functioning in schizophrenia]. AB - The major reviews of the literature support the idea that a significant proportion of patients with schizophrenia present cognitive deficits in several domains, more marked in the domains of verbal memory, vigilance and attention, memory, intellectual quotient, language and executive functioning. Such deficits appear to be one of the main determinants of these patients' functional outcome. More recently, social cognition deficits have been described. Social cognition may be understood as a separate and independent dimension of neurocognition or non-social cognition and may constitute a mediator between the neurocognition and functioning. However, there has been controversy concerning the real meaning of deficits observed due to the diversity of analysis methodologies employed and the fact that the available neuropsychological tests and batteries have not been specifically designed to evaluate cognitive deficits in patients with schizophrenia. In this paper, the Working Group on Schizophrenia (GTE) describes and highlights the existing clinical and scientific evidence, performs a critical review of cognitive functioning, social cognition and its impact on functional outcome, in patients with schizophrenia. The authors review definitions of (neuro)cognition, social cognition and functioning, analyze the existing methods for its assessment, describe the treatments available in this context and summarize the evidence of dysfunctions in these three concepts, taking into account their interconnection. Overall, the GTE considered the need for a standardized battery of tests to measure neurocognition, social cognition and functioning, consensually accepting the use of MATRICS as the standard tool for assessing neurocognition in schizophrenia. It was also recognized that verbal memory and vigilance deficits may be the best predictors of functional outcome in schizophrenia. In addition, the GTE has established social cognition as a priority area in the study of schizophrenia, however, the limitations in terminology and assessment methodologies do not allow a consensus in this area. The GTE considers that further longitudinal studies with larger samples are needed, so that a more adequate therapeutic armamentarium becomes available for patients with schizophrenia. PMID- 21627883 TI - [Ethical, technical and legal procedures of the medical doctor responsibility to accomplish the road enforcement law about driving under the influence of alcohol and psychotropic substances]. AB - The forensic toxicology (TF) is a science of analytical basis, aiming to clarify legal issues related to poisoning, whether or not fatal, within the various areas of law (criminal, civil, labor, etc.). The analysis that are more often requested (with a tendency to increase and gaining rising attention) are those concerning the procedures involving supervision of driving under the influence of alcohol and psychotropic substances, in the living individual and in the cadaver. The key players in this process, are: (a) the police agents carrying out the screening and quantification of alcohol on the exhaled breath and the screening of psychotropic and stupefacient substances in saliva; (b) the public health services that perform qualitative analysis of these substances in urine (if the test was not previously performed in saliva); (c) the doctor that collects blood samples from the living, or the dead victim; (d) the forensic toxicologist who conducts toxicological analysis in blood (or, eventually in another biological sample) and (e) the magistrate prosecutors that ultimately will receive the toxicological report to apply the law. Therefore it is important to understand and be acquainted with the road law enforcement of driving under the influence of alcohol and psychotropic substances, particularly in what concerns to the role of the medical doctor. Consequently, this paper aimed to review these topics, namely highlighting the necessary information to clarify the interested parties about the technical, ethical and legal procedures to consider. PMID- 21627884 TI - [Early intervention in psychosis: prepsychotic period]. AB - Psychotic disorders, namely schizophrenia, are severe illnesses with their onset in adolescence or youth adult age and have a classic limited outcome. In recent years there has been some research about early intervention in psychosis which can be done in prepsychotic period (prodrome) and/or after the onset of full blown psychotic episode. The authors review the literature about the diagnosis, evaluation, and possible interventions in patients in prepsychotic period (prodrome). Identification of patients with ultra high risk of psychosis could identify patients with a risk of development psychosis in 40%. The main interventions, pharmacological and psychosocial treatments, and main published studies are explained. Prospective identification of these patients and their treatment could prevent the development of full blown psychosis, delay the onset or promote the recovery of the disorder. We must remember some ethical issues about this kind of intervention, namely the stigma and false positive individuals. The results from the research in this area are mainly preliminary, but these studies suggest that some interventions are effective and promising. In future we must have trials with more number of patients included and probably a refinement of the criteria of ultra high risk patients. PMID- 21627885 TI - [Quality of life and transplantation]. AB - With the recent development of surgical techniques and other treatments of transplanted patients the increase in survival is not anymore the unique objective of the intervention. Nowadays, increase in quality of life is a very important aim. The instruments that assess quality of life can be multi/unidimensional, specific/nonspecific. One of the most important instruments to evaluate quality of life in all kinds of patients is the MOS-SF36, validated to Portuguese population. According to most published studies, there is an improvement in quality of life dimensions after transplantation. In some prospective studies it is shown that quality of life after transplantation is determined by some pre-transplantation factors such as medical factors (severity of illness) and psychiatric factors (personality, depression, coping strategies). PMID- 21627886 TI - [Depression in cancer patients: diagnostic and therapeutic considerations]. AB - BACKGROUND AND AIMS: Oncologic diseases currently have a high prevalence and present as one of the leading causes of death in the western world. Clinical depression and emotional distress are often the outcome of the threat these diseases present to individual existence. Although its precise determination is hampered by methodological problems, up to 50% of cancer patients may become clinically depressed and experience intense personal distress. We performed a literature review on screening and evaluating clinical depression in cancer patients (risk factors, instruments and strategies) and its treatment (psychotherapy, drug treatment and care management). METHODS: Non-systematic literature review. The search was performed on Pubmed with the following keywords in title/abstract fields: cancer, oncology, depression, psychiatry, morbidity, screening, treatment, psychotherapy, psychiatric status rating scales, with no time restriction. Articles written in Portuguese, English and Castilian were included. A cross-reference search yielded additional included articles. CONCLUSIONS: Several risk factors for an increased likelihood of clinical depression in oncologic diseases have been identified which, together with screening strategies, including validated scales such as Hospital Anxiety and Depression Scale (HADS), may enhance our ability to detect cases. While not having, for the moment, the highest possible evidence for effectiveness from randomized trials, the treatment strategies for clinical depression in this population should be available and make use of multidisciplinary interventions, including psychopharmacological and psychotherapeutic options. The need for health care workers to spend adequate time with patients is underscored. This not only enhances their ability to detect and treat depression cases but also allows for an empathic and understanding relationship, validating the existence and suffering of the patient. PMID- 21627887 TI - [Weber-Christian disease: unknown etiology systemic panniculitis]. AB - Panniculitis is an inflammation occurring within adipose tissue. This process is often associated with auto-immune diseases, infections, malignancy and others diseases of unknown cause. We report a 16-year-old woman with constitutional symptoms and relapsing nonsuppurative nodular panniculitis. The etiological study didn't identify any associated disease, on admission or in 2,5 years of follow up. This is a case of Weber-Christian disease with a favourable corticosteroid therapy result. The authors present a Weber-Christian disease bibliographic revision. PMID- 21627888 TI - [Gorlin-Goltz syndrome: review of the neuroradiological and maxillofacial features illustrated with two clinical cases]. AB - INTRODUCTION: Gorlin-Goltz syndrome or nevoid basal cell carcinoma syndrome is a rare hereditary autosomal-dominant disorder characterized by multiple basal cell carcinomas in young patients, odontogenic keratocysts, palmar or plantar pits, calcification of the falx cerebri and skeletal malformations. This syndrome is due to mutations in PTCH1 (patched homolog 1 da Drosophila), a tumor suppressor gene. Diagnostic criteria were defined by Evans, revised by Kimonis and include major and minor criteria. The authors review in particular the neuroradiological and maxillofacial characteristics of the syndrome. CASE REPORT: The authors describe the clinical presentation of two children with Gorlin-Goltz syndrome without affected first degree relatives. In both the clinical suspicion of the syndrome is raised by the presence of multiple odontogenic cysts surgically removed. Histopathological exam revealed keratocysts. None of the patients has basal cell carcinomas but both present with skeletal anomalies, namely marked pectus deformity. DISCUSSION/CONCLUSION: The absence of major diagnostic criteria like basal cell carcinomas or palmar or plantar pits in young patients delay the early diagnosis and the correct screening for medulloblastoma, basal cell carcinomas and cardiac fibromas. Odontogenic keratocysts are the most consistent clinical finding in Gorlin-Goltz syndrome in the first one or two decades of life. These patients are very sensitive to ionizing radiation, being able to develop basal cell carcinomas and meningiomas. Treatment should accomplish the complete resection of the tumors. PMID- 21627889 TI - [Bacterial endogenous endophthalmitis]. AB - Endogenous bacterial endophthalmitis is an uncommon disease, secondary to hematogenous spread of a septic focus. Despite advances in diagnostic and therapeutic methods, it remains a potential cause of blindness. We report a case of a diabetic patient with endogenous endophthalmitis of the right eye secondary to an Escherichia coli bacteremia of urinary origin. The evolution was torpid, with loss of visual acuity and evisceration of the right eye. PMID- 21627890 TI - [Chronic eosinophilic pneumonia]. AB - Idiopathic chronic eosinophilic pneumonia (ICEP) is a rare disorder of unknown cause, associated with intense and abnormal eosinophilic lung infiltration. Most cases occur in patients with prior history of atopy and asthma. It is characterized by subacute or chronic presentation, alveolar and blood eosinophilia and peripheral pulmonary infiltrates on chest imaging, which are sometimes migratory. All treated patients show good and rapid response to systemic corticosteroid therapy and favourable prognosis, however relapses are frequent after weaning or stopping treatment. By this reason, in about one-half of patients long-term systemic corticosteroid therapy is needed. We describe 2 clinical cases of ICEP which diagnosis was established after exclusion of any known cause of eosinophilic lung disease that represents different clinical, laboratorial and radiological patterns of this disease and are discussed in the context of the available data about ICEP and the differential diagnosis with other causes of pulmonary eosinophilia. PMID- 21627891 TI - [Endovascular thrombolysis for massive cerebral venous thrombosis in a teenager with nephrotic syndrome]. AB - Cerebral venous thrombosis is a rare but potentially severe condition in children. We present the case of a teenager with corticodependent nephrotic syndrome diagnosed at five months of age and treated with cyclosporine A. In the context of recurrence of nephrotic syndrome he presented with headache, vomiting and severe intracranial hypertension. While the raised intracranial pressure and the status epilepticus were controlled, the brain imaging revealed venous thrombosis of all venous sinus, with absence of venous drainage. He was submitted to local thrombolysis with recombinant tissue plasminogen activator, with recanalization of the venous sinuses. The outcome was favourable, without neurological deficits. In this case, the early radiologic intervention was crucial, enabling a full neurological recovery, in a teenager whose initial prognosis was very poor. PMID- 21627892 TI - [Osteopathia striata with cranial sclerosis]. AB - We report on a female patient who presented failure to thrive, laryngotracheomalacia, conductive deafness and facial dysmorphisms. A skeletal survey revealed thickening of the cranial vault, linear striations in the diametaphyses of all long bones and fan-like striations of the iliac bones. CT scan of the temporal bone showed thickening of the cranial base, sclerotic mastoids, abnormal ossicular fixation and stenosis of the otic foramina. The radiological findings led to the diagnosis of Osteopathia Striata with Cranial Sclerosis. A mutation in WTX gene confirmed the clinical and radiological diagnosis of Osteopathia Striata with Cranial Sclerosis in this patient and allowed proper genetic counseling and providing prenatal diagnosis. PMID- 21627893 TI - [Gastric heterotopia of rectum: a rare entity]. AB - Heterotopic gastric mucosa is an extremely rare condition; in the anorectal region only a very few cases were described up to now. The authors report a case of a 46-year-old woman, asymptomatic, to whom a massive recess was discovered in the rectum, at 12 cm of the anal verge; a bottom flat lesion with 1.5 cm of diameter revealed a focus of normal appearing fundic-type gastric mucosa at histology. PMID- 21627894 TI - [Neuroleptic malignant syndrome: a rare long lasting case report]. AB - Neuroleptic malignant syndrome (NMS) is a neurological emergency related to neuroleptics medication. Diagnosis is clinical. Authors present the case of a 32 year-old Caucasian man with a history of moderated oligophreny treated with stable doses of psychotropic and neuroleptic drugs. Because of aggressivity outbreaks, he was administered high doses of intramuscular haloperidol decanoate for a period of ten days. One month later he had evolved with mental deterioration, mutism, generalized rigidity, fever, tachycardia, hypotension and diaphoresis. Neuroleptic malignant syndrome was diagnosed. Supporting treatment was provided and intercurrences treated. Neuroleptic malignant syndrome is a fatal disease in 10 to 20% of patients; otherwise it resolves in about two weeks. In rare cases, like this one the authors report, it has a prolonged evolution of months. PMID- 21627896 TI - [Emergency admissions ward as place of education--can the expectations be fulfilled?]. PMID- 21627897 TI - [Clinical junior doctors' initial experiences with the joint interdisciplinary ward for acute admissions]. AB - A new hospital organisation for receiving acute patients has been introduced in Denmark. All patients with acute problems are to be received in an interdisciplinary ward (IW). This study examines how junior doctors evaluate training in the IW. A questionnaire was sent to 558 doctors who went through postgraduate basic training in 2009 and 2010. The response rate was 63%. Sixty eight doctors had been trained in IW. The responders found that training in IW had a high educational potential. They appreciated the broad training in acute conditions from several specialities. However, the educational management could be improved. PMID- 21627898 TI - [Statins can maybe reduce the risk of venous thromboembolism]. AB - Recent reports suggest beneficial effects of statin treatment on risk of venous thromboembolism (VTE). This review studies the existing literature on statins and VTE: one meta-analysis, one randomized controlled trial, four cohort studies and eight case-control studies. The hypothesis of a risk reducing effect of statins on recurrent VTE is supported, but the included studies are heterogeneous; and there is a risk of uncontrolled confounding. Randomized clinical trials aimed at clarifying whether statins can prevent recurrent VTE or not are warranted. PMID- 21627899 TI - [Complications following construction and closure of loop ileostomies]. AB - A protective loop ileostomy is used to reduce the incidence and consequences of anastomotic failure following colorectal resection. Closure of a loop ileostomy is associated with low mortality but many studies have demonstrated high morbidity rates. The aim of this review is to examine the existing evidence on the morbidity following closure of loop ileostomies and to investigate possible risk factors for complications. There is no consensus in the literature about the risk factors for complications. Counseling of the patient about the risks for complications is emphasized. PMID- 21627900 TI - [New diagnostic update in Acanthamoeba keratitis]. AB - Acanthamoeba keratitis is a rare but sight threatening condition. A major problem is that the disease is difficult to diagnose and often mistaken for herpes infection in its early stages. We present an update in diagnostics with the presentation of five recent cases of Acanthamoeba keratitis where the use of confocal microscopy and PCR based DNA analysis of epithelial scrapings played a pivotal role. An early diagnose is crucial for achieving a successful outcome. PMID- 21627901 TI - [Malignant tumours of the hand]. AB - Malignant tumours of the hand are rare and are often misdiagnosed. A painful swelling of the hand or digits are often diagnosed with an infection, benign tumours such as ganglion cysts, or arthritis. Wounds that do not heal despite adequate treatment should be biopsied to rule out malignancy. A correct diagnosis without delay is important because the life expectancy, due to a metastasis on the hand or fingers is approximately six months. PMID- 21627902 TI - [The child with chronic diarrhoea: suggestions for workup]. AB - Causes of chronic diarrhoea (CD) are numerous. At the first consultation it is possible by simple means to evaluate the child for common causes and screen for the more severe ones: infection, food allergy, disaccharide malabsorption, general malabsorption (celiac disease and cystic fibrosis), inflammatory bowel disease, and functional disorders (toddler's diarrhoea and irritable colon syndrome). Second line evaluation should take place in the paediatric ward, aiming to confirm suspected, complicated disorders and to explore the possibility of the large number of rare causes of CD in the seriously sick child. PMID- 21627903 TI - [Holistic-neuropsychological rehabilitation--documentation is lacking]. PMID- 21627904 TI - [Venovenous hemodiafiltration for patient with baclofen intoxication]. AB - Twice a young man was admitted to hospital upon having taken baclofen overdoses by intention. The ingested dose was 1,850 mg at the first episode and up to 2,500 mg at the second. In both cases the patient had severe overdose symptoms and scored 4-5 on the Glascow Coma Scale and was admitted to intensive care. Continuous venovenous hemodiafiltration (CVVHDF) was initiated. The elimination pharmacokinetics and toxicokinetics for baclofen is not fully known. During the second submission a shorter elimination half-life time was observed and it might be due to either compartmental distribution of baclofen, or more likely caused by an advantageous effect of the CVVHDF. PMID- 21627905 TI - [Internal hernia as a complication to laparoscopic gastric bypass]. AB - Internal hernia is a well-known complication after gastric bypass surgery. However, the diagnosis relies on a high degree of clinical suspicion and a low threshold for diagnostic laparoscopy as the results from clinical and radiological examination may be non-specific. We present two cases of internal herniation 24 months postoperatively with very different outcomes. PMID- 21627906 TI - [Cerebral infarctions in primary cardiac tumours]. AB - Primary heart neoplasms are rare and their clinical presentation can comprise classic cardiac symptoms, symptoms of embolism or constitutional signs. We describe the case of a 40 year-old woman with a primary heart neoplasm and discuss the epidemiological and clinical characteristics found in international literature. PMID- 21627907 TI - [Guidelines for type 2 diabetes can be discussed]. PMID- 21627908 TI - [Waiting lists shall be understood and organized correctly]. PMID- 21627909 TI - [Waiting time for treatment shall be calculated correctly]. AB - Every system is perfectly designed to achieve the results it gets and in the Danish health care system there is a significant waiting time for the patients. If we want this result to change, we have to change something in the system, but before we do that, we need to understand the system. With queuing theory as a frame of reference, the anatomy and physiology of waiting time is illuminated in order to bring additional knowledge into the design of systems in health care. PMID- 21627910 TI - [The haematological biobank--an infrastructure for quality assurance, development and research]. AB - Archived patient samples of biological material coupled to clinical databases are valuable and hold the potential to improve patient care, quality control, research and development. Biobanks form a catalytic infrastructure which ensures the registration, handling and storage of collected biological material. Here, we describe the Haematology Biobank and stress the significance of a registration database. Together, these form the basis for retrospective validation of clinical progress and prognostic evaluation of new diagnostic methods, which can therefore be more quickly implemented in the clinic. PMID- 21627911 TI - [Endophthalmitis is an acute disease which threatens the vision]. AB - Endophthalmitis is one of the most serious and most dreaded complications in ophthalmology. It is most often seen as a complication to intraocular surgery and especially after cataract surgery. Approximately 90% of the cases of endophthalmitis are caused by bacteria. Symptoms and signs may include pain, blurred vision, and hypopyon. Intravitreal injection of antibiotics in combination with vitrectomy or vitreous tap is the mainstay in the treatment of endophthalmitis. Prompt recognition and intervention are essential in preserving the vision of the affected eye in the case of endophthalmitis. PMID- 21627912 TI - [Intraosseous infusion]. PMID- 21627913 TI - [Arthrosis and arthroplastic surgery influence patients' sex life, depression, and socioeconomy]. AB - Over the past years there has been an increasing number of total hip or total knee arthroplastic operations. Due to demographic changes this tendency will probably continue. The operation frees the patient of disabling pain and stiffness and gives back the mobility to the patient. This systematic review addresses the impact an operation has on the patients' quality of life, economy, sex-life and possibly depression. We have identified relevant articles by search of PubMed MeSH database and a traditional search of PubMed. Furthermore we have evaluated the reference lists for articles of interest. PMID- 21627914 TI - [Ankylosing spondylitis is associated with increased cardiovascular morbidity and mortality]. AB - Ankylosing spondylitis is an inflammatory disorder primarily affecting the axial skeleton. The disease is associated with increased cardiovascular morbidity and mortality. Structural changes in the heart, and arteriosclerosis secondary to inflammation may be of importance. The role of traditional cardiovascular risk factors and of anti-inflammatory treatment is unclear. Tumor necrosis factor inhibitors seem to increase cholesterol levels. Evaluation of the cardiovascular risk in these patients should be considered. Cardiovascular risk factors should be managed according to ordinary guidelines. PMID- 21627915 TI - [A spontaneous dissection of cervical arteries]. AB - Presenting two cases of spontaneous dissection of cervical arteries. A 54 year old male suffered from left-sided Horner syndrome and severe headache. MR cerebrum revealed a left internal carotid artery dissection. A 57 year-old female with left vertebral artery dissection was admitted to hospital due to repeated attacks of total blindness preceded by weeks of moderate headache. PMID- 21627916 TI - [Leiomyoma can also be located in the vagina]. AB - Vaginal wall leiomyomas are rare. They are typically located in the anterior or lateral wall of the vagina. We report a case of a posterior vaginal leiomyoma which was mistaken for a cervical fibroma. Despite several examinations and biopsies to rule out a potential malignant condition, the diagnosis was delayed. Whenever there is a clinical suspicion of a vaginal leiomyoma, magnetic resonance imaging or translabial ultrasound are the recommended imaging modalities. PMID- 21627917 TI - [Unusual long-term survival following non-radical treatment of oral mucosal melanoma]. AB - Malignant mucosal melanomas are rare and aggressive. The five-year survival rate for malignant oral melanomas is 15%. A case of a 65-year old male with progressive melanosis in the oral mucus and development of four malignant melanomas during a 15-year follow-up period is presented. The patient was treated non-radically. Radical surgical excision of the melanotic area would have included partial mandibulectomy and maxillectomy followed by massive reconstruction and was thus not an option. The patient has been followed closely during the follow-up period and invasive components have been excised followed by local reconstruction. The patient remains without signs of metastatic disease. PMID- 21627918 TI - Blood transfusion practices: a little consistency please. PMID- 21627919 TI - Blood transfusion practice: a nationwide survey in Italy. AB - BACKGROUND: Blood transfusion is a complex activity. Some of the components of this activity are implementation of standard procedures, evaluation of appropriateness of the blood use, methods for bedside identification of patients and the release of blood in emergencies, during out-of-routine hours and from hospitals lacking a Blood Centre. An overview about how these issues are managed in Italy could be of interest. MATERIALS AND METHODS: A survey dealing with some issues regarding blood release was performed, using a questionnaire sent to 278 Italian Blood Centres. RESULTS: Out of the 278 Centres, 179 (64%) returned the questionnaire. The geographic distribution of the Centres that responded (80 in the north, 46 in the centre and 53 in the south of Italy) offers a good picture of the Italian reality. Globally there seems to be a relatively uniform application of procedures and guidelines, of methods to identify patients, and of evaluating transfusion appropriateness. The systems used to deal with emergency blood release and blood release in non-routine conditions are more variable. The use of technological resources seems to be disappointingly low. DISCUSSION: Although many aspects of the blood transfusion process should be improved, the picture that emerged from the survey seems to show, on the whole, a fair quality of blood transfusion practice in Italian Blood Centres. PMID- 21627921 TI - Anaerobic storage of red blood cells: the need for caution regarding donor red cells with sickle cell trait. PMID- 21627923 TI - Unwashed shed blood: should we transfuse it? PMID- 21627922 TI - Recommendations for the transfusion management of patients in the peri-operative period. III. The post-operative period. PMID- 21627924 TI - Red cell transfusion in orthopaedic surgery: a benchmark study performed combining data from different data sources. AB - BACKGROUND: Previous studies have shown the usefulness of combining information from different data sources to identify and analyse variations in transfusion practices. Good knowledge of the conditions leading to blood use is a fundamental requirement for the assessment of the appropriateness of blood transfusion. MATERIALS AND METHODS: In this study we combined blood transfusion data obtained from the Blood Bank information system with patients' data from the Hospital Discharge Database, based on the ICD9 classification system, from 1,827 surgical procedures performed in seven different orthopaedic divisions in the Ravenna area between January and December 2009. Hip and knee replacement surgery (primary or revision) and operations following femoral fractures (partial hip replacement and reduction with internal fixation) were considered. For a subgroup of patients clinical and transfusion data were also combined with haemoglobin values obtained from the laboratory information system. RESULTS: Of the 1,827 surgical procedures, 1,038 (56.8%) were followed by transfusion of red cells. The likelihood of receiving a transfusion varied depending on the patient's sex (49% for males, 60% for females), age, and on the surgical procedure, being higher for interventions following femoral fractures and for revisions of hip replacement: about 70% of patients undergoing these interventions required transfusion. A large variability in transfusion rates was observed between the seven divisions, which was only partially explained by the different types of surgery (post traumatic or elective) performed by any of them: relevant variations were also observed for the same type of intervention. DISCUSSION: Combining information from different data sources could be a time-sparing way to gain useful information about transfusion practices, so contributing to optimising blood usage. PMID- 21627925 TI - Factor VIII safety: plasma-derived versus recombinant products. PMID- 21627926 TI - Strategies for reducing transfusions in major orthopaedic surgery. PMID- 21627927 TI - Four phases of checks for exclusion of umbilical cord blood donors. AB - AIM: The aim of this study was to analyse umbilical cord blood (UCB) collection over 1 year between October 2008 and September 2009, seeking ways to improve the number of suitable banked UCB units. Four phases of the process were investigated, from the consent form to the banking procedure, paying attention to the discarded UCB units. MATERIAL AND METHODS: We recruited couples at 35 weeks of gestation and took an accurate history, focusing on genetic, immunological and infectious diseases. We collected UCB from pregnant women who delivered vaginally or by Caesarean section between the 37-41(+6) weeks of gestation. Some units were discarded on the basis of the patients' history, obstetric events or biological criteria. In utero collection was the preferred method of collection. RESULTS: During the study period, between October 2008 and September 2009, there were 1,477 deliveries in our unit. The number of couples interested in UCB donation was 595 (40.2%-595/1,477). We collected 393 UBC units. We excluded 122 patients at the phase of the history taking, counselling and informed consent (first phase check). Of the 393 units collected, 162 (41.3%) were banked whereas 231 (58.7%) were discarded because they did not fulfil biological criteria (third phase check). The volume of UCB units collected after Caesarean section was greater than the volume of units collected after vaginal delivery (95.4 mL versus 85.0 mL, respectively; p <0.01). The UCB units collected after vaginal delivery contained a higher number of total nucleated cells compared to the units collected after Caesarean section (970x10(6) cells versus 874x10(6) cells, respectively; p=0.037). None of the banked UCB units was discarded at the clinical check 6 months after delivery (fourth phase check). CONCLUSIONS: Our study shows that strict observance of each of the checks and the collection strategy is important to guarantee the safety of the UCB units and to maximise the cost-benefit ratio. After the appropriate checks we banked UCB units from only 27.2% (162/595) of the couples who gave consent to the procedure and from only 11% (162/1,477) of all the deliveries in the 12 month study period, as 59.8% of couples were not properly informed about UCB donation. PMID- 21627928 TI - The correlation between the granulocyte content in infused stem cells and side effects of the infusion. PMID- 21627929 TI - Evaluation of myocardial and hepatic iron loading by MRI T2* in multi-transfused patients with repeated blood loss as compared to thalassaemia major patients and controls. PMID- 21627930 TI - [Surgery of the transverse dimension]. PMID- 21627931 TI - [Surgery of the sagittal dimension]. PMID- 21627932 TI - [Surgery of the vertical dimension]. PMID- 21627933 TI - [Multidimensional surgery]. PMID- 21627934 TI - [Profile plastic surgery]. PMID- 21627935 TI - [Postoperative priorities]. PMID- 21627936 TI - [Conclusion]. PMID- 21627958 TI - The search for alternative aqueous film forming foams (AFFF) with a low environmental impact: physiological and transcriptomic effects of two Forafac((r)) fluorosurfactants in turbot. AB - Fluorosurfactants are the key components in aqueous film forming foams (AFFF). They provide these fire fighting agents with the required low surface tension and they enable film formation on top of lighter fuels to prevent burn back. Development of effective and environmentally acceptable PFOS alternatives is one of the most important priorities in the fire fighting foam industry. DuPontTM offers the fluorosurfactant mixtures Forafac((r))1157 and Forafac((r))1157N for the formulation of AFFFs which are alternatives to the persistent and toxic perfluorooctane sulphonate (PFOS). Ecotoxicological testing of these inadequately documented mixtures is necessary to include them in AFFF hazard and risk assessment. Juvenile turbot (Scophthalmus maximus) were exposed for 14 days to 0.1; 0.5 and 1.5mg/L of the fluorosurfactant mixtures used in Forafac((r))1157 and Forafac((r))1157N. In an initial transcriptomics experiment, microarray analysis revealed differentially expressed transcripts of genes which were mainly involved in digestion and in the immune system. This discovery-driven screening approach offered the basis for new hypotheses that were tested in two subsequent experiments in which food intake, energy reserves, growth and a set of haematological parameters were examined. Additionally, effects of the two mixtures were compared to those of PFOS. Based on the results of this study, the mode of action of Forafac((r))1157N was the activation of the acute phase reaction resulting in increased leukocyte concentrations and the inhibition of growth due to the high energetic cost of toxicant exposure. For Forafac((r))1157, evidences of immunosuppression were found on the transcriptional level and the altered differential leukocyte profiles indicated that stress was induced in these fish. However, food intake, energy reserves and growth were not compromised, even at high exposure concentrations, which was in contrast to the effects seen after PFOS exposure. Taking into account that Forafac((r))1157 appeared to be less toxic than PFOS, this mixture could be considered as a more environmentally acceptable PFOS alternative for the use in AFFFs. PMID- 21627959 TI - ROS enhances CXCR4-mediated functions through inactivation of PTEN in prostate cancer cells. AB - Inactivation of the tumor suppressor phosphatase and tensin homolog deleted on chromosome 10 (PTEN) is heavily implicated in the tumorigenesis of prostate cancer. Conversely, the upregulation of the chemokine (CXC) receptor 4 (CXCR4) is associated with prostate cancer progression and metastasis. Studies have shown that loss of PTEN permits CXCR4-mediated functions in prostate cancer cells. Loss of PTEN function is typically due to genetic and epigenetic modulations, as well as active site oxidation by reactive oxygen species (ROS); likewise ROS upregulates CXCR4 expression. Herein, we show that ROS accumulation permitted CXCR4-mediated functions through PTEN catalytic inactivation. ROS increased p-AKT and CXCR4 expression, which were abrogated by a ROS scavenger in prostate cancer cells. ROS mediated PTEN inactivation but did not affect expression, yet enhanced cell migration and invasion in a CXCR4-dependent manner. Collectively, our studies add to the body of knowledge on the regulatory role of PTEN in CXCR4 mediated cancer progression, and hopefully, will contribute to the development of therapies that target the tumor microenvironment, which have great potential for the better management of a metastatic disease. PMID- 21627960 TI - Protective effect of zinc against cadmium hepatotoxicity depends on this bioelement intake and level of cadmium exposure: a study in a rat model. AB - It was estimated, in a rat model of moderate and relatively high chronic human exposure to cadmium (Cd), whether enhanced zinc (Zn) consumption may prevent Cd induced liver injury and if the possible protective effect of this bioelement depends on its intake. For this purpose, the structure and function of the liver of the rats that received Zn (30 and 60mg/l) or/and Cd (5 and 50mg/l) for 6months were evaluated. The treatment with Cd led to, dependent on the exposure level, pathological changes in the liver, including enhanced apoptosis and induction of inflammatory and necrotic processes. Moreover, the serum activities of hepatic marker enzymes (alanine transaminase and aspartate transaminase) and the concentration of proinflammatory cytokine - tumor necrosis factor alpha were increased. The supplementation with 30 and 60mg Zn/l (enhancing daily Zn intake by 79% and 151%, respectively) partially or totally prevented from some of the Cd induced changes in the liver structure and function; however, it provided no protection from necrosis, and the administration of 60mg Zn/l during the higher Cd exposure even intensified this process. At both levels of Cd treatment, the use of 30mg Zn/l was more effective in preventing liver injury than that of 60mg Zn/l. The hepatoprotective impact of Zn may be explained, at least partly, by its antioxidative, antiapoptotic and anti-inflammatory action, ability to stimulate regenerative processes in the liver tissue, and indirect action resulting in a decrease in the liver pool of the non-metallothionein-bound Cd(2+) ions able to exert toxic action. The results provide strong evidence that enhanced Zn consumption may be beneficial in protection from Cd hepatotoxicity; however, its excessive intake at relatively high exposure to Cd may intensify liver injury. PMID- 21627961 TI - The ImmunoCAP ISAC molecular allergology approach in adult multi-sensitized Italian patients with respiratory symptoms. AB - OBJECTIVES: To evaluate the performances of an allergen microarray in multi sensitized allergic patients with respiratory symptoms. DESIGN AND METHODS: 321 patients and 92 controls were included in this study. Specific serum IgE were assayed using ImmunoCAP ISAC, a microarray containing 103 components derived from 47 allergens and results were compared with extract-based ImmunoCAP Allergens sIgE to 15 common airborne allergens. RESULTS: The reproducibility of ISAC was good. The Positive Percent Agreement (PPA) varied between 75% and 100% for sIgE levels above 1 kUA/l. For samples with sIgE levels below 0.1 kUA/l, the Negative Percent Agreement (NPA) ranged between 90% and 100%. Notably, 58% of respiratory allergy patients had IgE to food-specific proteins and 52% resulted sensitized to cross-reactive pan-allergens. CONCLUSION: ImmunoCAP ISAC detects allergen sensitization at component level and adds important information by defining both cross and co-sensitization to a large variety of allergen molecules. PMID- 21627962 TI - Influence of the preservation period in silica-gel on the predatory activity of the isolates of Duddingtonia flagrans on infective larvae of cyathostomins (Nematoda: Cyathostominae). AB - The continued maintenance of nematophagous fungi predatory activity under laboratory conditions is one of the basic requirements for a successful biological control. The purpose of this study was to evaluate the influence of time on the preservation of the fungus Duddingtonia flagrans (AC001 and CG722) stored in silica-gel for 7 years and their subsequent predatory activity on cyathostomin L(3) larvae in 2% water-agar medium (2% WA). Samples of the isolates AC001 and CG722, originating from vials containing grains of silica-gel sterilized and stored for 7 years, were used. After obtaining fungal conidia, the predation test was conducted over 7 days on the surface of 9.0 cm Petri dishes filled with 2% WA. In the treated groups each Petri dish contained 500 cyathostomin L(3) and conidia of fungal isolates in 2% WA. In the control group (without fungi) the plates contained 500 L(3) in 2% WA. The experimental results showed that isolated AC001 and CG722 were efficient in preying on cyathostomin L(3) (p<0.01) compared to control (without fungus). However, no difference was observed (p>0.01) in the predatory activity of the fungal isolates tested. Comparing the groups, there was a significant reductions of cyathostomin L(3) (p<0.01) of 88.6% and 78.4% on average recovered from the groups treated with the isolates AC001 and CG722, respectively, after 7 days. The results of this test showed that the fungus D. flagrans (AC001 and CG722) stored in silica-gel for at least 7 years maintained its predatory activity on cyathostomin L(3). PMID- 21627963 TI - Pathogenesis of restricted movements in trichinellosis: an experimental study. AB - Trichinellosis is a zoonosis acquired by the ingestion of insufficiently cooked pork meat containing the encapsulated larvae of Trichinella spiralis. Trichinellosis is presented with myalgia which affects various muscle groups; its intensity is usually related to the severity of the disease and may cause restriction of joint movement. However, joint pain in the course of trichinellosis could not be explained entirely by myositis. This study investigated the other possible causes of restricted movements of joints in animal model. We found that the histopathological changes in the joints of T. spiralis infected rats were in the form of inflammatory cellular infiltrates and ulceration in the synovial membrane with degeneration and ulceration of the articular cartilage. Immunohistochemical examination of the joints revealed the presence of T. spiralis local antigen or immune complex deposited in the synovial membrane. Leukocytosis and eosinophilia were observed throughout the experimental period but eosinophil level declined slowly but still elevated. In conclusion, the restricted movements during the course of trichinellosis seem to be not only due to direct invasion of muscles by the encapsulated T. spiralis larvae but also due to immune complex deposition in the joints. PMID- 21627964 TI - Mucosal immunity against Eimeria acervulina infection in broiler chickens following oral immunization with profilin in MontanideTM adjuvants. AB - The present study was conducted to compare aqueous nanoparticle-based MontanideTM IMS 1313 N VG PR (IMS 1313) and oil-based ISA 71 VG (ISA71) adjuvants in combination with an Eimeria subunit protein vaccine on protection against avian coccidiosis. Male broiler chicks were vaccinated twice with an Eimeria recombinant profilin protein alone or in conjunction with IMS 1313 or ISA 71 prior to infection with live, sporulated Eimeria acervulina oocysts. For comparison, chickens were immunized with a commercial live coccidiosis vaccine (Coccivac-B). The following parameters were assessed as measures of protective immunity: body weight gain, fecal oocyst output, profilin-specific intestinal secretary IgA (sIgA) or IgY antibody levels, and percentages of CD4(+), CD8(+), TCR1(+), or TCR2(+) intestinal intraepithelial lymphocytes (IELs). Birds vaccinated with profilin plus ISA 71 had increased body weight gains, equivalent to Coccivac-B vaccination, compared with the profilin-only group. Immunization with profilin plus IMS 1313, or with profilin plus ISA 71, reduced fecal oocysts shedding compared with the profilin-only group. Intestinal sIgA levels were greater in the profilin plus IMS 1313 or ISA 71 groups, and IgY levels were greater in the profilin plus ISA 71 group, compared with profilin alone. Birds vaccinated with profilin plus IMS 1313 or ISA 71 had higher percentages of CD4(+), CD8(+), and TCR1(+), but not TCR2(+), intestinal IELs compared with the profilin-only vaccinated group. Taken together, these results indicate that immunization of chickens with the recombinant profilin subunit vaccine in conjunction with IMS 1313 or ISA 71 adjuvants increases protective immunity against experimental E. acervulina infection. PMID- 21627965 TI - Trichinella spiralis: the influence of short chain fatty acids on the proliferation of lymphocytes, the goblet cell count and apoptosis in the mouse intestine. AB - This study was carried out to determine the influence of short chain fatty acids (SCFA) on spleen and mesenteric lymph node lymphocyte proliferation, goblet cells and apoptosis in the mouse small intestine during invasion by Trichinella spiralis. BALB/c mice were infected with 250 larvae of T. spiralis. An SCFA water solution containing acetic, propionic and butyric acids (30:15:20 mM) was administered orally starting 5 days before infection and ending 20 days post infection (dpi). Fragments of the jejunum were collected by dissection 7 and 10 dpi, and were examined for apoptotic cells in the lamina propria of the intestinal mucosa, and for goblet cells. The proliferation index of the cultured spleen and mesenteric lymph node lymphocytes with MTT test was also determined. The orally administered SCFA solution decreased the proliferation of mesenteric lymph node lymphocytes in the mice infected with T. spiralis at both examination times, but did not influence the proliferative activity of the spleen cells. Seven dpi, both in the spleen and mesenteric lymph nodes, the highest proliferation index of concanavalin A (Con A)-stimulated lymphocytes was found in the group of uninfected animals receiving SCFA animals. This tendency could still be seen 10 dpi in the mesenteric lymph nodes but not in the spleen, where the proliferation index in this group had significantly decreased. In vitro studies revealed, that butyric and propionic acids added to the cell cultures suppressed the proliferation of Con A-stimulated mesenteric lymph nodes and spleen lymphocytes taken from uninfected and T. spiralis-infected mice. Acetic acid stimulated proliferation of splenocytes taken from uninfected mice but did not affect lymphocyte proliferation in mesenteric lymph nodes from uninfected or infected mice. Orally administered SCFA increased the number of goblet cells found in the epithelium of the jejunum 7 dpi, but this number had decreased 10 dpi. The number of apoptotic cells in the lamina propria of the intestinal mucosa of animals infected with the T. spiralis and receiving SCFA was also lower, particularly 10 dpi. The above results show that SCFA can participate in the immune response during the course of trichinellosis in mice. PMID- 21627966 TI - Energetic metabolism of axenic promastigotes of Leishmania (Viannia) braziliensis. AB - Leishmania spp are protozoans capable of carbohydrates degradation and as energy source they can use glucose, aminoacids or lipids from the environment. The products of the metabolic pathways such as organic acids may be used as an index of their energetic metabolic profile. Therefore, in this study a metabolic profile comparison was made between promastigotes from one reference strain (MHOM/BR/1975/M2903) and two different isolates of Leishmania (Viannia) braziliensis (MHOM/BR/2003/IMG3 and MHOM/BR/2005/RPL5). The parasites culture was performed in complete Grace's culture media seeded in 24-well plates at 26 degrees C. During the growth curve performance samples were collected from the logarithmic and stationary phases of culture and therefore analyzed by high performance liquid chromatography (HPLC) and spectrophotometry assays to determine the concentrations of glucose, lactate, citrate, alpha-ketoglutarate, succinate, fumarate, malate, oxaloacetate and beta-hydroxybutirate which are indicative of the energetic pathways. It was possible to detect an increase in the glucose from the stationary phase from the M2903 strain when compared to the logarithmic phase while in the IMG3 and RPL5 isolates there was a decrease (p<0.05). The spectrophotometric and chromatographic results indicated that the logarithmic phase which presents higher energy consumption due to the intense replication rate have the energetic pathways intensified. It was also possible to note some metabolic differences between the analyzed parasites which may indicate possible adaptations of the parasite when facing different environmental and physiological conditions during its life cycle and that these differences may help in the understanding of the diversity of the host-parasite relationship from Leishmania parasites. PMID- 21627967 TI - Folding kinetic pathway of phosphofructokinase-2 from Escherichia coli: a homodimeric enzyme with a complex domain organization. AB - Phosphofructokinase-2 is a 66 kD homodimer whose subunits are associated by means of a bimolecular domain, the beta-clasp, which is linked to the larger portion of each subunit by a reentrant chain topology. To investigate how this structural organization determines the folding pathway of Pfk-2, unfolding and folding kinetic experiments were performed. The folding pathway shows an unstructured monomeric intermediate and that most part of the dimer structure is reached as a slow concerted folding/association step with a quite folded transition state in terms of solvent exposure. Unfolding kinetics show a transient intermediate, probably a partially unfolded dimer. We propose that these characteristics arise by a mutual constrain between the large domain and the beta-clasp domain imposed by their interrupted chain connectivity. PMID- 21627968 TI - Structure-activity relationships influencing lipid-induced changes in eIF2alpha phosphorylation and cell viability in BRIN-BD11 cells. AB - Fatty acids influence the viability of eukaryotic cells differentially such that long chain saturated molecules are poorly tolerated, whereas unsaturated species are less detrimental and can be cytoprotective. The basis for these effects is unclear but studies in yeast imply that they reflect the spatial configuration of the molecules when incorporated into the ER membrane. Using BRIN-BD11 beta-cells, we show that a wide range of unsaturated free fatty acids and their methyl-esters (having differing chain length and disposition of the double bonds) elicit cytoprotection and relief of protein kinase RNA-like endoplasmic reticulum kinase dependent ER stress. Thus, both physical properties and specific signalling events may regulate fatty acid responses in beta-cells. PMID- 21627969 TI - TRADD is critical for resistance to TRAIL-induced cell death through NF-kappaB activation. AB - One major obstacle in the clinical application of TRAIL as a cancer therapeutic agent is the acquisition of TRAIL resistance. We found that deficiency of TRADD sensitizes cells to TRAIL-induced apoptosis. Enhanced cell death in TRADD(-/-) MEFs is associated with defective NF-kappaB activation, indicating that the pro survival function of TRADD in TRAIL signaling is mediated at least in part via NF kappaB activation. Moreover, siRNA knock-down of TRADD in cancer cells sensitizes them to TRAIL-induced apoptosis. Thus, TRADD has a survival role in TRAIL signaling and may be one potential target for overcoming TRAIL resistance in cancer therapy. PMID- 21627970 TI - The Mg2+ transporter MagT1 partially rescues cell growth and Mg2+ uptake in cells lacking the channel-kinase TRPM7. AB - Magnesium (Mg(2+)) transport across membranes plays an essential role in cellular growth and survival. TRPM7 is the unique fusion of a Mg(2+) permeable pore with an active cytosolic kinase domain, and is considered a master regulator of cellular Mg(2+) homeostasis. We previously found that the genetic deletion of TRPM7 in DT40 B cells results in Mg(2+) deficiency and severe growth impairment, which can be rescued by supplementation with excess extracellular Mg(2+). Here, we show that gene expression of the Mg(2+) selective transporter MagT1 is upregulated in TRPM7(-/-) cells. Furthermore, overexpression of MagT1 in TRPM7(-/ ) cells augments their capacity to uptake Mg(2+), and improves their growth behavior in the absence of excess Mg(2+). PMID- 21627971 TI - Mitochondrial density contributes to the immune response of macrophages to lipopolysaccharide via the MAPK pathway. AB - We investigated the role of mitochondrial reactive oxygen species (ROS) in the response of macrophages to lipopolysaccharide (LPS) using RAW 264.7 cells and their rho(o) cells lacking mitochondria. Mitochondrial density, respiratory activity and related proteins in rho(o) cells were significantly lower than those in RAW cells. LPS rapidly stimulated mitochondrial ROS prior to cytokine secretion, such as TNF-alpha and IL-6, from RAW 264.7 cells by activating the MAPK pathway, while the response was attenuated in rho(o) cells. Exposure of rho(o) cells to H(2)O(2) partially restored the secretion of cytokines induced by LPS. These results suggest that mitochondrial density and/or the respiratory state contribute to intracellular oxidative stress, which is responsible for the stimulation of LPS-induced MAPK signaling to enhance cytokine release from macrophages. PMID- 21627972 TI - Inhibition of NF-kappaB by MG132 through ER stress-mediated induction of LAP and LIP. AB - Proteasome inhibitor MG132 blocks activation of NF-kappaB by preventing degradation of IkappaB. In this report, we propose an alternative mechanism by which MG132 inhibits cytokine-triggered NF-kappaB activation. We found that MG132 induced endoplasmic reticulum (ER) stress, and attenuation of ER stress blunted the suppressive effect of MG132 on NF-kappaB. Through ER stress, MG132 up regulated C/EBPbeta mRNA transiently and caused sustained accumulation of its translational products liver activating protein (LAP) and liver-enriched inhibitory protein (LIP), both of which were identified as suppressors of NF kappaB. Our results disclosed a novel mechanism underlying inhibition of NF kappaB by MG132. PMID- 21627973 TI - Translation of a histone H3 tail as a model system for studying peptidyl-tRNA drop-off. AB - We found that the synthesis of histone H3 N-terminal peptide (tail) in a reconstituted protein synthesis system yielded fragmented peptides along with the full-length product. With the combined use of MALDI-TOF analysis and peptidyl tRNA hydrolase cleavage of the Flag tagged product species, we concluded that the fragments were generated by peptidyl-tRNA drop-off at specific sites and subsequent translation continuation. Using the histone H3 tail we also found that peptidyl-tRNA drop-off is strongly correlated with the amino acid context. We envision that the system described here would be useful as a model system for studying peptidyl-tRNA drop-off events. PMID- 21627974 TI - Effects of simulated microgravity on brain plasticity: a startle reflex habituation study. AB - There is limited but increasing evidence that space environment, namely weightless condition, may affect astronauts' cerebral neurotransmitters and cognitive performance. The present experiment hypothesized that learning and brain plasticity are affected by simulated microgravity condition. To this aim, 22 male subjects matching astronauts' characteristics were divided in two groups, Head-Down Bed Rest (HDBR) and Sitting Control. After 3-h bed rest (or sitting condition) subjects started a picture viewing task during which 30 acoustic startle probes (100 dBA loudness), divided into three consecutive blocks, were delivered through headphones while startle reflex amplitude was measured from the EMG of the orbicularis oculi muscle. Habituation analysis of the startle reflex showed a normal reflex inhibition across blocks in sitting controls and no habituation in HDBR subjects. Results point to a microgravity-induced lack of startle reflex plasticity in subjects matching astronauts, a learning deficit which may affect the success of long-term space missions. PMID- 21627975 TI - Obesity: the allostatic load of weight loss dieting. AB - The obesity epidemic that is prevailing in most countries of the world is generally attributed to the increased amount of opportunities to be in positive energy balance in a context of modernity. This obviously refers not only to sedentariness and unhealthy eating that may dominate life habits of many individuals but also to unsuspected non-caloric factors which produce discrete allostatic changes in the body. In this paper, the focus is put on the impact of some of these factors with the preoccupation to document the allostatic burden of weight loss. Thus, beyond the fact that modernity favors opportunities to eat much and not to be active, the proposed conceptual integration leads to the conclusion that a modern lifestyle makes weight loss more difficult for obese individuals. In addition to the natural effects of weight loss favoring resistance to lose fat, a lifestyle promoting shorter sleep duration and more cognitive demand produces allostatic changes that may interfere with weight loss. The case of persistent organic pollutants (POPs) is also discussed as an example of the potential detrimental effects of a contaminated environment on metabolic processes involved in the control of energy expenditure. Taken together, these observations suggest that weight loss is more than ever a search for compromise between its metabolic benefits and its allostatic effects promoting body weight regain. PMID- 21627976 TI - Disease-specific adverse events following nonlive vaccines: a paradoxical placebo effect or a nocebo phenomenon? AB - Vaccines can cause adverse reactions (AR), i.e. adverse events following immunization (AEFIs) due to the vaccine, such as local reactions or fever. In addition, live attenuated vaccines which replicate in vaccinees can cause disease specific AR, e.g. measles-like rash following measles vaccination. However, nonlive vaccines because they are inactivated and they do not replicate in vaccinees, are not likely to cause disease-specific AR. The aim of the study was to assess whether safety signals could be generated by an undescribed bias in spontaneous reporting of disease-specific AEFIs with nonlive vaccines. All AEFIs of Sanofi Pasteur MSD vaccines spontaneously reported in France from January 2000 to June 2010, coded according to MedDRA terms and collected in the company's pharmacovigilance database were analyzed. Vaccine-event pairs of interest were selected a priori. The disproportionality reporting rate methodology was used, comparing the proportion of a given event reported following a given vaccine to its proportion reported following all other studied vaccines. The Reporting Odds Ratio (ROR) was used for signals detection for each vaccine-event pair selected. A total of 33,275 AEFIs were analyzed. The calculated ROR showed a statistically disproportionate reporting rate and generated false safety signals for almost all the pairs tested. Three nonlive vaccine pairs were striking: gynaecological symptoms and the quadrivalent human papillomavirus (qHPV) vaccine; trismus and tetanus vaccines; hepatobiliary disorders and hepatitis B vaccines. In conclusion we have identified a new vaccine AE spontaneous reporting bias: "disease-specific adverse events following nonlive vaccines", showing that vaccinees and healthcare professionals tend to report preferentially the symptoms of the disease against which the nonlive vaccine was administered. We suggest that bias is subordinate to a paradoxical placebo effect and/or a nocebo phenomenon. PMID- 21627977 TI - Soluble interleukin-2 receptor levels correlated with positive symptoms during quetiapine treatment in schizophrenia-spectrum disorders. AB - BACKGROUND: Some but not all antipsychotics have been shown to modulate plasma cytokine levels in schizophrenia patients. Thus far, the most consistent finding has been the increase in plasma levels of soluble interleukin (IL)-2 receptor (sIL-2R) associated with clozapine treatment. Quetiapine is a second-generation antipsychotic with a pharmacological profile similar to that of clozapine, but its immunomodulatory effects have not been investigated in schizophrenia yet. The purpose of this exploratory study was to examine the changes in plasma levels of sIL-2R in schizophrenia during quetiapine treatment and association with psychopathology. METHODS: Participants were 29 schizophrenia-spectrum disorder patients (DSM-IV criteria), and 28 healthy controls. Patients had a comorbid substance use disorder (cannabis>alcohol>cocaine), since quetiapine is increasingly used in this population of dual diagnosis. No participant suffered from infection or overt inflammatory diseases. On baseline, patients taking mostly second-generation antipsychotics were switched to quetiapine for a 12-week open-label trial. Five patients were drop-outs. Mean dose of quetiapine for trial completers (n=24) was 466.6mg+/-227.3. Psychiatric variables were evaluated with the Positive and Negative Syndrome Scale and the Calgary Depression Scale for Schizophrenia. Plasma sIL-2R levels were assessed at baseline, weeks 6 and 12 in patients, and in healthy controls, using sandwich immunoassay. Plasma IL-6 and IL 1 receptor antagonist (IL-1RA) were measured for comparison purposes. RESULTS: On baseline, plasma sIL-2R, IL-6 and IL-1RA levels were higher in dual-diagnosis patients, compared to controls. Plasma sIL-2R further increased after quetiapine treatment (p=0.037), while plasma IL-6 and IL-1RA did not change. Clinical improvements were observed in positive, negative and depressive symptoms, and substance abuse severity (all p<0.01). Interestingly, changes in sIL-2R levels during treatment were inversely correlated with changes in positive symptoms (r= 0.524; p=0.009). That is, increases in sIL-2R levels were associated with reductions in positive symptoms. CONCLUSION: These data show that quetiapine elevates, like clozapine, sIL-2R levels in schizophrenia. Furthermore, the results suggest that sIL-2R alterations in schizophrenia rely on complex interplays between antipsychotics and the positive symptoms of the disorder. Future randomized controlled trials involving larger samples of schizophrenia patients are warranted to determine whether changes in plasma sIL-2R are quetiapine-related. PMID- 21627978 TI - Abnormal mitoferrin-1 expression in patients with erythropoietic protoporphyria. AB - OBJECTIVE: Most patients with erythropoietic protoporphyria have deficient ferrochelatase (FECH) activity due to changes in FECH DNA. We evaluated seven patients with erythropoietic protoporphyria phenotype in whom abnormalities of FECH DNA were not found by conventional analysis. The major focus was mitoferrin 1 (MFRN1), the mitochondrial transporter of Fe used for heme formation by FECH and for 2Fe2S cluster synthesis, which is critical to FECH activity/stability. MATERIALS AND METHODS: Four patients had a deletion in ALAS2 that causes enzyme gain-of-function, resulting in increased formation of protoporphyrin; one had a heterozygous major deletion in FECH DNA. All had an abnormal transcript of MFRN1 in messenger RNA extracted from blood leukocytes and/or liver tissue. The abnormal transcript contained an insert of intron 2 that had a stop codon. The consequences of abnormal MFRN1 expression were examined using zebrafish and yeast MFRN-deficient strains and cultured lymphoblasts from the patients. RESULTS: Abnormal human MFRN1 complementary DNA showed loss-of-function in zebrafish and yeast mutants, whereas normal human MFRN1 complementary DNA rescued both. Using cultured lymphoblasts, quantitative reverse transcription polymerase chain reaction showed increased formation of abnormal transcript that was accompanied by decreased formation of normal transcript and reduced FECH activity in patients compared to normal lines. A positive correlation coefficient (0.75) was found between FECH activity and normal MFRN1 messenger RNA in lymphoblasts. However, no obvious cause for increased formation of abnormal transcript was identified in MFRN1 exons and splice junctions. CONCLUSIONS: Abnormal MFRN1 expression can contribute to erythropoietic protoporphyria phenotype in some patients, probably by causing a reduction in FECH activity. PMID- 21627979 TI - Management of postural sensory conflict and dynamic balance control in late-stage Parkinson's disease. AB - Parkinson's disease (PD) is known to affect postural control, especially in situations needing a change in balance strategy or when a concurrent task is simultaneously performed. However, few studies assessing postural control in patients with PD included homogeneous population in late stage of the disease. Thus, this study aimed to analyse postural control and strategies in a homogeneous population of patients with idiopathic advanced (late-stage) PD, and to determine the contribution of peripheral inputs in simple and more complex postural tasks, such as sensory conflicting and dynamic tasks. Twenty-four subjects with advanced PD (duration: median (M)=11.0 years, interquartile range (IQR)=4.3 years; Unified Parkinson's Disease Rating Scale (UPDRS): M "on dopa"=13.5, IQR=7.8; UPDRS: M "off-dopa"=48.5, IQR=16.8; Hoehn and Yahr stage IV in all patients) and 48 age-matched healthy controls underwent static (SPT) and dynamic posturographic (DPT) tests and a sensory organization test (SOT). In SPT, patients with PD showed reduced postural control precision with increased oscillations in both anterior-posterior and medial-lateral planes. In SOT, patients with PD displayed reduced postural performances especially in situations in which visual and vestibular cues became predominant to organize balance control, as was the ability to manage balance in situations for which visual or proprioceptive inputs are disrupted. In DPT, postural restabilization strategies were often inefficient to maintain equilibrium resulting in falls. Postural strategies were often precarious, postural regulation involving more hip joint than ankle joint in patients with advanced PD than in controls. Difficulties in managing complex postural situations, such as sensory conflicting and dynamic situations might reflect an inadequate sensory organization suggesting impairment in central information processing. PMID- 21627980 TI - Distinct intrinsic membrane properties determine differential information processing between main and accessory olfactory bulb mitral cells. AB - Most mammals rely on semiochemicals, such as pheromones, to mediate their social interactions. Recent studies found that semiochemicals are perceived by at least two distinct chemosensory systems: the main and accessory olfactory systems, which share many molecular, cellular, and anatomical features. Nevertheless, the division of labor between these systems remained unclear. Previously we suggested that the two olfactory systems differ in the way they process sensory information. In this study we found that mitral cells of the main and accessory olfactory bulbs, the first brain stations of both systems, display markedly different passive and active intrinsic properties which permit distinct types of information processing. Moreover, we found that accessory olfactory bulb mitral cells are divided into three neuronal sub-populations with distinct firing properties. These neuronal sub-populations can be integrated in a simulated neuronal network that neglects episodic stimuli while amplifying reaction to long lasting signals. PMID- 21627981 TI - Potential role for S100A4 in the disruption of the blood-brain barrier in collagen-induced arthritic mice, an animal model of rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is an autoimmune disease associated with chronic inflammation of the joints. RA has been shown to increase the morbidity of and mortality due to cardiovascular and cerebrovascular diseases. We recently reported that cerebrovascular permeability was increased in mice with collagen induced arthritis (CIA), an animal model of RA. S100A4, a member of the S100 family, is up-regulated in synovial fluid and plasma from RA patients. This study was aimed at evaluating a role of S100A4 in the mediation of blood-brain barrier (BBB) dysfunction in CIA mice. CIA was induced by immunization with type II collagen in mice. Cerebrovascular permeability was assessed by measurement of sodium fluorescein (Na-F) levels in the brains of control and CIA mice. Serum S100A4 concentrations in control and CIA mice were measured by enzyme-linked immunosorbent assays (ELISA). Accumulation of Na-F in the brain and serum levels of S100A4 were increased in CIA mice. Increased S100A4 levels in the serum are closely correlated with hyperpermeability of the cerebrovascular endothelium to Na-F. We investigated whether S100A4 induces BBB dysfunction using mouse brain capillary endothelial cells (MBECs). S100A4 decreased the transendothelial electrical resistance and increased Na-F permeability in the MBECs. S100A4 reduced the expression of occludin, a tight junction protein, and stimulated p53 expression in MBECs. These findings suggest that S100A4 increases paracellular permeability of MBECs by decreasing expression levels of occludin, at least in part, via p53. The present study highlights a potential role for S100A4 in BBB dysfunction underlying cerebrovascular diseases in patients with RA. PMID- 21627982 TI - Vagus nerve stimulation modulates cortical synchrony and excitability through the activation of muscarinic receptors. AB - Vagus nerve stimulation (VNS) is an FDA approved treatment for drug-resistant epilepsy and depression. Recently, we demonstrated the capacity for repeatedly pairing sensory input with brief pulses of VNS to induce input specific reorganization in rat auditory cortex. This was subsequently used to reverse the pathological neural and perceptual correlates of hearing loss induced tinnitus. Despite its therapeutic potential, VNS mechanisms of action remain speculative. In this study, we report the acute effects of VNS on intra-cortical synchrony, excitability, and sensory processing in anesthetized rat auditory cortex. VNS significantly increased and decorrelated spontaneous multi-unit activity, and suppressed entrainment to repetitive noise burst stimulation at 6-8 Hz but not after application of the muscarinic antagonist scopolamine. Collectively, these experiments demonstrate the capacity for VNS to acutely influence cortical synchrony and excitability and strengthen the hypothesis that acetylcholine and muscarinic receptors are involved in VNS mechanisms of action. These results are discussed with respect to their possible implications for sensory processing, neural plasticity, and epilepsy. PMID- 21627983 TI - Opposite modulation of time course of quantal release in two parts of the same synapse by reactive oxygen species. AB - Reactive oxygen species (ROS) are potent regulators of transmitter release in chemical synapses, but the mechanism of this action remains almost unknown. Presynaptic modulation can change either the release probability or the time course of quantal release, which was recently recognized as an efficient mechanism determining synaptic efficiency. The nonuniform structure and a big size of the frog neuromuscular junction make it a useful model to study the action of ROS in compartments different in release probability and in time course of transmitter release. The time course (or kinetics) of quantal release could be estimated by measuring the dispersion of the synaptic delays for evoked uniquantal endplate currents (EPCs) under low release probability. Using two electrode recording technique, the action of ROS on kinetics and release probabilities were studied at the proximal and distal parts within the same neuromuscular junction. The stable ROS hydrogen peroxide (H2O2) increased the dispersion of synaptic delays of EPCs (i.e. desynchronized quantal release) within the distal part but decreased delay dispersion (synchronized quantal release) within the proximal part of the same synapse. Unlike the opposite modulation of kinetics, H2O2 reduced release probability in both distal and proximal parts. Since ATP is released from motor nerve terminals together with acetylcholine and can be involved in ROS signaling, we tested the presynaptic action of ATP. In the presence of the pro-oxidant Fe2+, extracellular ATP, similarly to H2O2, induced significant desynchronization of release in the distal regions. The antioxidant N-acetyl-cysteine attenuated the inhibitory action of ATP on release probability and abolished the action of H2O2 and ATP in the presence of Fe2+, on release kinetics. Our data suggest that ROS induced during muscle activity could change the time course of transmitter release along the motor nerve terminal to provide fine tuning of synaptic efficacy. PMID- 21627984 TI - Experiments and numerical modeling to estimate the coating variability in a pan coater. AB - The purpose of this work is to investigate the effect of the coating process parameters on coating performance and coating variability, and hence determine the optimal operating conditions. Coating of particles is done to mask the unpleasant taste or odor of the drug, to control the bioavailability of the API, and to increase shelf-life. The coating solution is sprayed in specific locations of the granular bed and coating uniformity is achieved by interparticle collisions and overall mixing behavior in the coater. Thus, good understanding of particle flow and granular mixing in a pan coater is vital to optimize the process parameters to reduce coating variability. Coating experiments are performed at previously determined optimal mixing conditions using Lactose nonpareils. The coating fluid (aqueous solution of Opadry II) is sprayed intermittently at different flow rates and concentration. Vernier Caliper is used to measure the change in diameter and the coating of the particles. Moreover, DEM based numerical modeling of spray coating is also performed for same operational parameter set and spray characteristic (center and the radius of the spray zone) used in the experiments. DEM simulation provides the residence time distribution of all the particles passing through the spray zone. The coating variability in the experiments is estimated at different pan and spray variables. The coating variability decreases with the increase inpan tilt, coating time and an optimum speed. The spray characteristics does not seem to have much effect on the variability although better coating is observed under better mixing conditions of high tilt and pan speed for the same spray parameters. The mass distribution of coated particles is quantified in the numerical model by the total number of particles passing through the spray zone and also by the frequency distribution of the residence time of the coated particles. It is observed that the simulations are in good agreement with the experiments for the effect of orientation (tilt) of the pan coater on coating variability. However simulations over predicted the effect of speed as compared to the experiments to reach the minimum coating variability. In the current study, the experimental setup did not reflect the typical bead coating setup used in the industry; rather depict a simplified setup to validate the numerical model. PMID- 21627985 TI - The stability of solid dispersions of felodipine in polyvinylpyrrolidone characterized by nanothermal analysis. AB - Nanothermal analysis (NTA) supported by atomic force microscopy imaging has been used to study the changes that occur at the surfaces of solid dispersions of the drug felodipine and the water soluble polymer, polyvinylpyrrolidone (PVP) on exposure to standard pharmaceutical environmental stress conditions. Exposure to relative humidities above 75% (at 40 degrees C) was sufficient to achieve phase separation of the drug and polymer into areas which displayed a glass transition temperature consistent with pure drug and polymer over a period of a few days. Higher values of humidity at 25 degrees C (e.g. 95%RH) were also sufficient to cause such phase separation within a day. Extended studies of up to two months showed an eventual crystallization of the drug. NTA is shown to be effective at the early detection of instabilities in solid dispersions and the quantifiable identification of the relative composition of phase separated domains based upon their glass transition temperatures. The combined nanoscale analytical approach employed here is able to systematically study the influence of storage conditions and different drug loadings and to evaluate physical stability as a function of environmental conditions. PMID- 21627986 TI - Plant use in the medicinal practices known as "strict diets" in Chazuta valley (Peruvian Amazon). AB - AIM OF THE STUDY: Strict diets are traditional medicinal practices where plant remedies are consumed with nearly fasting and with some sort of social seclusion. The aim of this work was to describe these practices of Chazuta and the use of plants within, as well as to analyse the possible functions of the last. MATERIAL AND METHODS: The information was obtained through interviews to the 6.3% of the district rural adult population (140 individuals, 75% of which was considered Quechua). RESULTS: In total, 122 strict diets were recorded and 106 different plant species were reported to be used. Strict diets present a characteristic structure and plant use. The main effects reported in strict diets were antinflammatory, antiinfective, brain function alteration and depuration. CONCLUSIONS: Strict diets are well structured traditional medicinal practices, also with a symbolic significance in the life cycle of chazutian men. Plants used in strict diets can contribute to the main effects through antinflammation, antiinfective actions, psychoactivity and depurative related activities. The correlation between literature evidence of activity of most used plants and effects reported for the correspondent diet (i.e. in which the plant was used) are 36% for antinflammatory activity, 29% for antimicrobial activity, 18% for psychoactivity and 5% for depurative related activities. The percentages go to 77%, 64%, 73% and 32%, respectively, when literature evidences on related taxa are also considered. PMID- 21627989 TI - Evaluation of water-in-oil-in-water multiple emulsion and microemulsion as potential adjuvants for immunization with rabies antigen. AB - Water-in-oil-in-water (W/O/W) multiple emulsions and microemulsions have been studied as potential candidates to be formulated as adjuvants. In this work their application as adjuvants for rabies virus immunization was studied. The humoral response, the effective dose 50 and histology for the developed formulations were evaluated in mice and compared with those from traditional adjuvants. The microemulsion and W/O/W multiple emulsion adjuvants developed were able to induce humoral response in mice and the serum showed good in vivo protection. Compared to the other adjuvants evaluated, microemulsion was shown to be the best candidate for rabies immunization as it presented good potency against the virus and did not appear to cause any local reaction. PMID- 21627988 TI - Hyaluronan-binding protein 1 (HABP1/p32/gC1qR) induces melanoma cell migration and tumor growth by NF-kappa B dependent MMP-2 activation through integrin alpha(v)beta(3) interaction. AB - Cell migration is the hallmark of cancer regulating anchorage independent growth and invasiveness of tumor cells. Hyaluronan (HA), an ECM polysaccharide is shown to regulate this process. In the present report, we demonstrated, supplementation of purified recombinant hyaluronan binding protein 1(HABP1/p32/gC1qR) from human fibroblast cDNA enhanced migration potential of highly invasive melanoma (B16F10) cells. Exogenous HABP1 adhered to the cell surface transiently and was shown to interact and colocalize with alpha(v)beta(3) integrin, a regulatory molecule of cell migration. In HABP1 treated cells, the phosphorylation of nuclear factor inducing kinase (NIK) and IkappaBalpha was observed, followed by nuclear translocation of p65 subunit of NFkappaB, along with its DNA-binding and transactivation, resulting in upregulation of MT1-MMP expression and finally MMP 2 activation. To substantiate our findings, prior to HABP1 treatment, the expression of NIK was reduced by small interfering RNA mediated knockdown and confirmed the inhibition of nuclear translocation of p65 subunit of NFkappaB and upregulation of MT1-MMP expression. In addition, the use of curcumin, an anti cancer drug, or GRGDSP, the blocking peptide along with exogenous HABP1, inhibited such NFkappaB-dependent pathway, confirming that HABP1-induced cell migration is alpha(v)beta(3) integrin-mediated and downstream signaling by NFkappaB. Finally, we translated the in vitro data in mice model and observed enhanced tumor growth with higher MT1-MMP expression and MMP-2 activation in the tumors upon injection of HABP1 treated melanoma cells. The treatment of curcumin, the anticancer drug along with HABP1, inhibited the migration, expression of MT1 MMP and activation of MMP-2 and finally tumor growth supports the involvement of HABP1 in tumor formation. PMID- 21627990 TI - Neuronal trafficking of voltage-gated potassium channels. AB - The computational ability of CNS neurons depends critically on the specific localization of ion channels in the somatodendritic and axonal membranes. Neuronal dendrites receive synaptic inputs at numerous spines and integrate them in time and space. The integration of synaptic potentials is regulated by voltage gated potassium (Kv) channels, such as Kv4.2, which are specifically localized in the dendritic membrane. The synaptic potentials eventually depolarize the membrane of the axon initial segment, thereby activating voltage-gated sodium channels to generate action potentials. Specific Kv channels localized in the axon initial segment, such as Kv1 and Kv7 channels, determine the shape and the rate of action potentials. Kv1 and Kv7 channels present at or near nodes of Ranvier and in presynaptic terminals also influence the propagation of action potentials and neurotransmitter release. The physiological significance of proper Kv channel localization is emphasized by the fact that defects in the trafficking of Kv channels are observed in several neurological disorders including epilepsy. In this review, we will summarize the current understanding of the mechanisms of Kv channel trafficking and discuss how they contribute to the establishment and maintenance of the specific localization of Kv channels in neurons. PMID- 21627991 TI - Loss of Thr286 phosphorylation disrupts synaptic CaMKIIalpha targeting, NMDAR activity and behavior in pre-adolescent mice. AB - In order to provide insight into in vivo roles of CaMKIIalpha autophosphorylation at Thr286 during postnatal development, behavioral, biochemical, and electrophysiological phenotypes of pre-adolescent Thr286 to Ala CaMKIIalpha knock in (T286A-KI) and WT mice were examined. T286A-KI mice displayed cognitive deficits in a novel object recognition test and an anxiolytic phenotype in the elevated plus maze, suggesting disruption of normal developmental processes. At the molecular level, the ratio of total CaMKIIalpha to CaMKIIbeta in hippocampal lysates was significantly decreased~2-fold in T286A-KI mice, and levels of both isoforms in synaptic subcellular fractions were decreased by~80%. Total levels of GluA1 AMPA-glutamate receptor subunits and phosphorylation of GluA1 at the CaMKII site (Ser831) in synaptic fractions were unaltered, as were the frequency and amplitude of AMPAR-mediated spontaneous excitatory postsynaptic currents at hippocampal CA3-CA1 synapses. Synaptic levels of NMDA-glutamate receptor GluN1, GluN2A and GluN2B subunits also were unaltered. However, the reduced ratio of CaMKII to NMDAR subunits in synaptic fractions was linked to increased synaptic NMDAR-mediated currents in T286A-KI mice, apparently due to increased functional contributions by GluN2B NMDARs (assessed by Ro 25-6981 sensitivity). Thus, disruption of CaMKII synaptic targeting caused by elimination of Thr286 autophosphorylation leads to synaptic and behavioral deficits during pre adolescence. PMID- 21627987 TI - Reactive oxygen species in cardiovascular disease. AB - Based on the "free radical theory" of disease, researchers have been trying to elucidate the role of oxidative stress from free radicals in cardiovascular disease. Considerable data indicate that reactive oxygen species and oxidative stress are important features of cardiovascular diseases including atherosclerosis, hypertension, and congestive heart failure. However, blanket strategies with antioxidants to ameliorate cardiovascular disease have not generally yielded favorable results. However, our understanding of reactive oxygen species has evolved to the point at which we now realize these species have important roles in physiology as well as pathophysiology. Thus, it is overly simplistic to assume a general antioxidant strategy will yield specific effects on cardiovascular disease. Indeed, there are several sources of reactive oxygen species that are known to be active in the cardiovascular system. This review addresses our understanding of reactive oxygen species sources in cardiovascular disease and both animal and human data defining how reactive oxygen species contribute to physiology and pathology. PMID- 21627992 TI - Limiting factors in atomic resolution cryo electron microscopy: no simple tricks. AB - To bring cryo electron microscopy (cryoEM) of large biological complexes to atomic resolution, several factors--in both cryoEM image acquisition and 3D reconstruction--that may be neglected at low resolution become significantly limiting. Here we present thorough analyses of four limiting factors: (a) electron-beam tilt, (b) inaccurate determination of defocus values, (c) focus gradient through particles, and (d) particularly for large particles, dynamic (multiple) scattering of electrons. We also propose strategies to cope with these factors: (a) the divergence and direction tilt components of electron-beam tilt could be reduced by maintaining parallel illumination and by using a coma-free alignment procedure, respectively. Moreover, the effect of all beam tilt components, including spiral tilt, could be eliminated by use of a spherical aberration corrector. (b) More accurate measurement of defocus value could be obtained by imaging areas adjacent to the target area at high electron dose and by measuring the image shift induced by tilting the electron beam. (c) Each known Fourier coefficient in the Fourier transform of a cryoEM image is the sum of two Fourier coefficients of the 3D structure, one on each of two curved 'characteristic surfaces' in 3D Fourier space. We describe a simple model-based iterative method that could recover these two Fourier coefficients on the two characteristic surfaces. (d) The effect of dynamic scattering could be corrected by deconvolution of a transfer function. These analyses and our proposed strategies offer useful guidance for future experimental designs targeting atomic resolution cryoEM reconstruction. PMID- 21627993 TI - Mussel collagen molecules with silk-like domains as load-bearing elements in distal byssal threads. AB - Mechanically stressed biological materials like tendon, spider silk or mussel byssal threads are typically composite materials comprising multi-domain proteins, in which molecular building blocks contribute to overall material function. Mussel byssal threads are the anchorage of sessile mytilid mussels, which withstand recurring external loads from waves and tides. A single thread is elastic and ductile proximally, while the distal portion exhibits an extraordinary stiffness and toughness with a transient gradient of both mechanical features along the thread. The main components of byssal threads include a set of various collagen-like structural proteins (preCols) consisting of a collagenous core sequence flanked by globular domains. Here, structural analysis using polarized Fourier-transform infrared spectroscopy (FTIR) on stretched distal portions of mussel byssal threads determines the impact of external linear load on various molecular moieties. It is concluded that the preCol collagenous core domain is the main load-bearing element in distal byssal threads, while polyalanine beta-sheets in the flanking domains, similar to those found in spider silk proteins, provide high stiffness at low strains. Load dissipation is mediated by domain stretching of amorphous glycine-rich helical moieties followed by complete unfolding of the preCol flanking domains. PMID- 21627994 TI - Species delimitation in taxonomically difficult lichen-forming fungi: an example from morphologically and chemically diverse Xanthoparmelia (Parmeliaceae) in North America. AB - Mounting evidence suggests many morphology-based species circumscriptions in lichenized ascomycetes misrepresent fungal diversity. The lichenized ascomycete genus Xanthoparmelia includes over 800 described species displaying a considerable range of morphological and chemical variation. Species circumscriptions in this genus have traditionally been based on thallus morphology, medullary chemistry, and the presence or absence of sexual or asexual reproductive structures. Notwithstanding concerted effort on the part of taxonomists to arrive at a natural classification, modern taxonomic concepts for the most part remain unclear. Here we assess the evolution of characters traditionally regarded as taxonomically important by reconstructing a phylogenetic hypothesis based on sequence data from four nuclear ribosomal markers as well as fragments from two protein-coding nuclear loci. A total of 414 individuals were tested, representing 19 currently accepted species. Most sampled species, as currently circumscribed, were recovered as polyphyletic, suggesting that major diagnostic characters have evolved in a homoplasious manner. The vagrant growth form, distinct medullary chemistries, and production of vegetative diaspores appear to have evolved independently multiple times. Application of a population assignment test resulted in the recognition of 21 species-level genetic clusters, each of which was supported by a comparison of genetic distances as well as a Bayesian species delimitation method calculating probabilities associated with speciation events. Inferred clusters are largely incongruent with traditionally circumscribed species due to the prevalence of cryptic diversity and, in some cases, high levels of intraspecific morphological and chemical variation. These results call for a major taxonomic revision of Xanthoparmelia species in western North America. PMID- 21627995 TI - Speciation at the Mogollon Rim in the Arizona Mountain Kingsnake (Lampropeltis pyromelana). AB - Studies of speciation and taxon delimitation are usually decoupled. Combining these methods provides a stronger theoretical ground for recognizing new taxa and understanding processes of speciation. Using coalescent methods, we examine speciation, post-speciation population demographics, and taxon delimitation in the Arizona Mountain Kingsnake (Lampropeltis pyromelana), a species restricted to high elevations in southwestern United States and northern Mexico (SW). These methods provide a solid foundation for understanding how biogeographic barriers operate at the regional scale in the SW. Bayesian species delimitation methods, using three loci from samples of L. pyromelana taken throughout their range, show strong support for the existence of two species that are separated by low elevation habitats found between the Colorado Plateau/ Mogollon Rim and the Sierra Madre Occidental. Our results suggest an allopatric mode of speciation given the near absence of gene flow over time, which resulted in two lineages of unequal population sizes. Speciation likely occurred prior to the Pleistocene, during the aridification of the SW and/or the uplift of the Colorado Plateau, and while these species occupy similar high-elevation niches, they are isolated by xeric conditions found in the intervening low deserts. Furthermore, post speciation demographics suggest that populations of both lineages were not negatively impacted by climate change throughout the Pleistocene. Finally, our results suggest that at least for this group, where divergence is old and gene flow is low, Bayesian species delimitation performs well. PMID- 21627996 TI - Bioenergetics and intestinal phenotypic flexibility in the microbiotherid marsupial (Dromiciops gliroides) from the temperate forest in South America. AB - The microbiotherid marsupial Dromiciops gliroides inhabits the temperate forests of the Southern hemisphere, facing seasonal nutritional and energetic bottlenecks due to its apparently facultative insectivory/frugivory. In order to understand the physiological processes behind this ecological pattern, we studied the morpho physiological changes that D. gliroides exhibits after dietary acclimation, in a sample of 21 wild-caught individuals fed over 1 month with ad libitum diet of: (1) fruit, (2) insects or (3) a mix of insects and fruit. In addition, we measured oxygen consumption (VO(2)) at resting conditions. We also performed enzyme assays (sucrase, maltase, trehalase and aminopeptidase N) and measurements of organ morphology. We found that D. gliroides cannot fulfil its nutrient requirements only from insects or fruit. It needs a mixed diet in order to maintain its body mass and energy balance. However, as a response of diet acclimation, individuals showed several-fold changes in the activities of aminopeptidase-N, maltase and sucrase (but not trehalase). This result, both the magnitude of change and the simultaneous effects on three enzymes suggests that D. gliroides could exhibit adaptive phenotypic plasticity in the activity of intestinal enzymes. This study suggests also that D. gliroides, the only living representative of the Microbiotheria order, exhibits physiological adaptations to a generalist diet. PMID- 21627997 TI - Effects of divergent selection for yolk testosterone content on growth characteristics of Japanese quail. AB - Effects of yolk androgens on postnatal growth of offspring have been widely studied but their physiological role in the growth control is not fully understood due to an inconsistency in obtained results. We investigated androgen mediated maternal effects on postnatal growth in relation to endocrine control mechanisms using two lines of Japanese quail divergently selected for high (HET) and low (LET) egg testosterone (T) content. Embryonic growth did not differ between the lines. During the growth period HET quail were heavier and displayed longer tarsi as compared with LET quail, with more pronounced line differences in males than females. HET males were heavier than LET males from the age of 2 weeks, reached the age of maximum growth rate earlier, and displayed higher asymptotic body weight than LET males. Accelerated growth in HET males was not accompanied by increased postembryonic plasma T concentrations. Plasma triiodothyronine levels did not differ between lines while plasma thyroxine levels were decreased in HET as compared with LET female chicks. Line differences in body weight disappeared in adult quail suggesting that yolk androgens, increased in a physiological way, resulted in stimulation of juvenile growth rate in precocial Japanese quail under stable social and environmental conditions. PMID- 21627998 TI - Limitations of tpi and bg genes sub-genotyping for characterization of human Giardia duodenalis isolates. AB - The intestinal protozoan Giardia duodenalis includes 2 genetically distinct assemblages, A and B, which are responsible for human infections. Little is known so far on the genotypes of G. duodenalis human isolates in France. The present characterization of 19 French clinical isolates was aimed at determining their genotype patterns and associations with clinical symptoms, and in vivo metronidazole resistance, respectively. Based on both triose-phosphate isomerase (tpi) and beta-giardin (bg) gene sequences, twelve isolates were identified as assemblage A, and 7 as assemblage B for the 2 gene loci. Sub-genotyping heterogeneities were observed in 15/19 isolates attributed to either A or B assemblage. They include frequent mismatches and intra-assemblage discordances and mixed positions, which were found more frequently in tpi than in bg sequences, and in assemblage B than in assemblage A sequences. No association was found between sub-genotypes, clinical symptoms and metronidazole sensitivity. Present data underline the need for improvements in the standardization of G. duodenalis multilocus genotyping approach for further molecular epidemiologic studies of giardiasis. PMID- 21627999 TI - Cirrhosis with refractory ascites: serial large volume paracentesis, TIPS, or transplantation? PMID- 21628001 TI - A community-based clinical trial of Intra-Venous zOledRonic acid once Yearly in comparison to oral bisphosphonates in postmenopausal women with osteoporosis: the IVORY trial Methodological considerations. AB - BACKGROUND: Zoledronic acid is an intravenous once yearly bisphosphonate that has been shown to be effective and safe in improving BMD (bone mineral density) and reducing fracture risk in controlled clinical trials. IVORY is a Canadian post marketing study aiming at assessing real-life effectiveness, health care resource utilization, safety and compliance to treatment with zoledronic acid in comparison to orally administered bisphosphonates (OBP). METHODS: IVORY is a prospective two cohort observational study of patients treated with zoledronic acid or OBP. Eligible patients are postmenopausal females, >45 years old with osteoporosis for whom initiation of treatment with OBP or zoledronic acid is indicated. Subjects will be followed for four years. Outcomes are the change in lumbar spine, femoral neck and total hip BMD and the incidence of fractures. The study cohort will consist of 920 patients treated with zoledronic acid and 460 treated with OBP. Additional comparisons will be based on external standardization to the population of Quebec patients treated with OBP. DISCUSSION: Post Marketing Observational Studies (PMOS) are essential for the assessment of real-life effectiveness and population based benefit-risk ratios. The effect of access to care, compliance, adherence to guidelines, patient comorbidity and concomitant medication use could only be assessed with observational studies. IVORY will provide information about true life effectiveness, benefit-risk ratios, cost-effectiveness and barriers to the process-outcome optimization. The results will have implications for decision makers and health care stakeholders regarding the management of osteoporosis in Canada. PMID- 21628000 TI - Chronic liver disease in the Hispanic population of the United States. AB - Chronic liver disease is a major cause of morbidity and mortality among Hispanic people living in the United States. Environmental, genetic, and behavioral factors, as well as socioeconomic and health care disparities among this ethnic group have emerged as important public health concerns. We review the epidemiology, natural history, and response to therapy of chronic liver disease in Hispanic patients. The review covers nonalcoholic fatty liver disease, viral hepatitis B and C, coinfection of viral hepatitis with human immunodeficiency virus, alcoholic cirrhosis, hepatocellular carcinoma, autoimmune hepatitis, and primary biliary cirrhosis. For most of these disorders, the Hispanic population has a higher incidence and more aggressive pattern of disease and overall worse treatment outcomes than in the non-Hispanic white population. Clinicians should be aware of these differences in caring for Hispanic patients with chronic liver disease. PMID- 21628002 TI - Live imaging of endogenous Collapsin response mediator protein-1 expression at subcellular resolution during zebrafish nervous system development. AB - Collapsin response mediator proteins (CRMPs) are cytosolic phosphoproteins that are functionally important during vertebrate development. We have generated a zebrafish gene trap line that produces fluorescently tagged Crmp1 protein, which can be dynamically tracked in living fish at subcellular resolution. The results show that Crmp1 is expressed in numerous sites in the developing nervous system. Early expression is apparent in the forebrain, epiphysis, optic tectum and the developing spinal cord. In the larval brain, Crmp1 is expressed in several distinct brain regions, such as the telencephalon, habenula and cerebellum. In addition, it is expressed in the spinal cord in a manner that persists in the larva. The results suggest that this Crmp1 protein trap line offers a powerful tool to track selected neuronal populations at high resolution. PMID- 21628003 TI - MyD88-dependent Toll-like receptor signaling is required for murine macrophages response to IS2. AB - IS2, a soluble beta-glucan isolated from the cell wall of mutated Saccharomyces cerevisiae (S. cerevisiae) enhances the immune response compared to the wild type (WT) beta-glucan. In the present investigation we report that Toll-like receptor (TLR)/MyD88 signaling pathway was responsible in IS2 beta-glucan-mediated cellular response in RAW264.7 murine macrophages. Data revealed that IS2 beta glucan significantly up-regulated the TLR2/TLR4 expression. Moreover, TLR2/TLR4 responds to IS2 resulting in murine macrophage activation. In addition, the IS2 signal led to cytokine secretions of IL-6 and TNF-alpha. In the case of thioglycolate-elicited peritoneal macrophages from MyD88-deficient mice, the decrease in cytokines was observed. Further the mitogen-activated protein kinases (MAPKs) phosphorylation was evident and degradation of IkappaB-alpha was increased after stimulation with IS2 beta-glucan. Further examination with MyD88 deficient mice revealed that the MyD88 pathway might play an important role for IS2 beta-glucan-mediated activation of macrophages. PMID- 21628004 TI - Autotransplantation of circulating endothelial progenitor cells protects against lipopolysaccharide-induced acute lung injury in rabbit. AB - Acute lung injury and acute respiratory distress syndrome (ALI/ARDS) are leading causes of morbidity and mortality in critically ill patients. Recent studies suggest that endothelial progenitor cells (EPCs) transplantation could become a novel cell-based therapeutic strategy for ALI/ARDS, but the exact therapeutic effect and possible mechanisms still need to be elucidated. In the present study, autologous circulating EPCs were obtained from rabbits using Ficoll centrifugation and cultured in vitro for 7 days. ALI was induced in rabbits by lipopolysaccharide (LPS), and EPCs were administered systemically. Fluorescence microscopy showed that CM-DiI labelled EPCs could migrate to the injured lung tissues. Reduced pulmonary edema level, inflammation, hemorrhage and hyaline membrane formation were present in rabbit treated with EPCs. EPCs autotransplantation significantly decreased the expression of adhesion molecules of sICAM-1 and P-selectin. Furthermore, EPCs administration mediated a down regulation of proinflammatory responses (reducing IL-1beta and TNF-alpha) while increasing the anti-inflammatory cytokine IL-10. Apoptosis of endothelial and epithelial cells was substantially reduced in EPCs-treated rabbit. Those findings suggest that autotransplantation of circulating EPCs can reduce the severity of LPS-induced ALI. Possible mechanisms include EPCs engraftment and reendothelization, down-regulation of adhesion molecules, alleviation of inflammatory response and apoptosis prevention. PMID- 21628005 TI - Characteristics of effective Internet-mediated interventions to change lifestyle in people aged 50 and older: a systematic review. AB - Worldwide, the number of people aged 60 years and older steadily grows to a predicted 2 billion in 2050. Online interventions increasingly target lifestyle risk factors to promote healthy aging. The objective of this systematic review is to evaluate whether Internet mediated lifestyle interventions can successfully change lifestyle in people aged 50 and older. A PubMed search was conducted resulting in twelve articles, based on ten studies. The studies focused on physical activity, weight loss, nutrition, and diabetes. Nine studies used feasible interventions, with an average small to moderate effect size. The most important result is that there are multiple studies reporting positive lifestyle changes in an older population. On average, complex interventions, whether they present tailored or generic information, and online or offline comparison, are more effective than interventions with only one component. Internet mediated interventions hold great potential in implementing effective lifestyle programs, capable of reaching large populations of older persons at very low costs. PMID- 21628007 TI - Fibromyalgia syndrome: a discussion of the syndrome and pharmacotherapy. PMID- 21628008 TI - Multisociety guideline on reprocessing flexible gastrointestinal endoscopes: 2011. PMID- 21628009 TI - Management of ingested foreign bodies and food impactions. PMID- 21628010 TI - Devices to improve colon polyp detection. PMID- 21628011 TI - Stents in the proximal esophagus: tailoring the stent to the patient to achieve success. PMID- 21628012 TI - Esophageal cancer patients undergoing external beam radiation after placement of self-expandable metal stents: is there a risk of radiation dose enhancement? AB - BACKGROUND: Self-expandable metal stents (SEMSs) are used for palliation of malignant dysphagia. It is not known whether dose adjustments are required when there is a stent in the radiation field. OBJECTIVE: To measure the effects of esophageal stents of various designs and materials on radiation dose to the tissue adjacent to the stent in the radiation field to determine whether there should be any dose adjustment. DESIGN: Simulated clinical protocol. SETTING: Linear accelerator radiation treatment center. PATIENTS: Solid Water phantoms were used to mimic the tissue environment of the human esophagus as well as stents of various designs and materials and controls. INTERVENTIONS: Radiation beams composed of photons (x-rays) delivered in split dosing with energies of 6, 10, and 15 million volts. MAIN OUTCOME MEASUREMENTS: Film and image-based evidence of dose enhancement; Monte Carlo calculations. RESULTS: Dose enhancement from single beams was seen only on the anterior surface, particularly in the stainless steel Z-stent (3.5%-7.8%) and the nonmetal Polyflex stent (5.5%-8.8%); less dose enhancement was seen on the anterior surface of the Alimaxx and Ultraflex nitinol stents (2%-2.5%). A negligible dose effect was seen on the posterior wall of all the stents tested. Monte Carlo calculation results were roughly similar to actual dosimeter measurements. LIMITATIONS: Simulated clinical protocol. CONCLUSIONS: This tissue-mimicking model reveals that radiation dose enhancement is a function of stent design and material, and the dose reduction is unnecessary as long as multiple fields are used. PMID- 21628013 TI - Idiopathic recurrent pancreatitis: an EUS-based management approach. PMID- 21628014 TI - Prospective, randomized comparison of two small-bowel capsule endoscopy systems in patients with obscure GI bleeding. AB - BACKGROUND: Video capsule endoscopy is the first-intention examination in patients with obscure GI bleeding. The new MiroCam capsule, when using electric field propagation for transmission, has been poorly evaluated in a clinical setting, in contrast with the PillCam SB2 capsule. OBJECTIVE: To evaluate the diagnostic concordance (kappa value) between PillCam SB2 and MiroCam capsule examinations performed in the same patients. DESIGN AND SETTING: Prospective, randomized study in 7 endoscopy units. PATIENTS AND INTERVENTION: Eighty-three consecutive patients, ingesting the 2 capsules at a 1-hour interval. RESULTS: Seventy-three patients were analyzed (10 technical issues). There were 31 concordant negative cases (42.4%) and 30 concordant positive cases (41.1%). The study showed satisfactory diagnostic concordance between the 2 systems (kappa = 0.66). In 12 patients (16.4%), the final diagnosis was different: 9 patients had positive findings on MiroCam examination but no image detected with PillCam SB2, 2 had positive findings on PillCam examination only, and 1 patient had 2 different diagnoses. A positive diagnosis was obtained in 46.6% and 56.2% of patients with PillCam SB2 and MiroCam capsule, respectively, so that the procedures identified 78.6% and 95.2% of positive cases, respectively (P = .02). Small-bowel transit time and capsule reading time were significantly longer in MiroCam procedures. LIMITATIONS: Technical failures possibly related to capsule interference. CONCLUSION: This study shows at least comparable efficiency of the MiroCam compared with the PillCam SB2 capsule system for the diagnosis of obscure GI bleeding. PMID- 21628015 TI - Development and validation of a novel method for assessing competency in polypectomy: direct observation of polypectomy skills. AB - BACKGROUND: Despite its ubiquitous use over the past 4 decades, there is no structured, formal method with which to assess polypectomy. OBJECTIVE: To develop and validate a new method with which to assess competency in polypectomy. DESIGN: Polypectomy underwent task deconstruction, and a structured checklist and global assessment scale were developed (direct observation of polypectomy skills [DOPyS]). Sixty bowel cancer screening polypectomy videos were randomly chosen for analysis and were scored independently by 7 expert assessors by using DOPyS. Each parameter and the global rating were scored from 1 to 4 (scores >=3 = competency). The scores were analyzed by using generalizability theory (G theory). SETTING: Multicenter. RESULTS: Fifty-nine of the 60 videos were assessable and scored. The majority of the assessors agreed across the pass/fail divide for the global assessment scale in 58 of 59 (98%) polyps. For G-theory analysis, 47 of the 60 videos were analyzed. G-theory analysis suggested that DOPyS is a reliable assessment tool, provided that it is used by 2 assessors to score 5 polypectomy videos all performed by 1 endoscopist. DOPyS scores obtained in this format would reflect the endoscopist's competence. LIMITATIONS: Small sample and polyp size. CONCLUSIONS: This study is the first attempt to develop and validate a tool designed specifically for the assessment of technical skills in performing polypectomy. G-theory analysis suggests that DOPyS could reliably reflect an endoscopist's competence in performing polypectomy provided a requisite number of assessors and cases were used. PMID- 21628016 TI - Bowel preparation with split-dose polyethylene glycol before colonoscopy: a meta analysis of randomized controlled trials. AB - BACKGROUND: Polyethylene glycol (PEG) is a commonly used bowel preparation for colonoscopy. Unfortunately, the standard large-volume solution may reduce patient compliance. Split-dosing of PEG has been studied in various randomized, controlled trials (RCTs). However, results have been conflicting. OBJECTIVE: We conducted a meta-analysis to assess the role of split-dose PEG versus full-dose PEG for bowel preparation before colonoscopy. DESIGN: Multiple databases were searched (January 2011). RCTs on adults comparing full-dose and split-dose of PEG for bowel preparation before colonoscopy were included and analyzed by calculating pooled estimates of quality of bowel preparation, preparation compliance, willingness to repeat the same preparation, and side effects by using odds ratio (OR) by fixed and random-effects models. SETTING: Literature search. PATIENTS: Per RCTs. MAIN OUTCOME MEASUREMENTS: Satisfactory bowel preparation, willingness to repeat same bowel preparation, patient compliance, and side effects. RESULTS: Five trials met inclusion criteria (N = 1232). Split-dose PEG significantly increased the number of satisfactory bowel preparations (OR 3.70; 95% CI, 2.79-4.91; P < .01) and willingness to repeat the same preparation (OR 1.76; 95% CI, 1.06-2.91; P = .03) compared with full-dose PEG. Split-dose PEG also significantly decreased the number of preparation discontinuations (OR 0.53; 95% CI, 0.28-0.98; P = .04) and nausea (OR 0.55; 95% CI, 0.38-0.79; P < .01) compared with full-dose PEG. LIMITATIONS: Limited number of studies. CONCLUSIONS: The use of a split-dose PEG for bowel preparation before colonoscopy significantly improved the number of satisfactory bowel preparations, increased patient compliance, and decreased nausea compared with the full-dose PEG. PMID- 21628017 TI - ERCP: pearls of wisdom from master endoscopists. PMID- 21628018 TI - An endoscopist's guide to political involvement: because inaction is no longer a reasonable alternative. PMID- 21628019 TI - High-resolution and optical molecular imaging for the early detection of colonic neoplasia. PMID- 21628020 TI - Spondylodiscitis complicating cholangitis caused by stent occlusion. PMID- 21628021 TI - Air cholangiography for severe hilar obstruction at ERCP. PMID- 21628022 TI - Propofol in upper GI endoscopy in patients with cirrhosis. PMID- 21628024 TI - Covered biliary metal stent: which are worse--the concepts, current models, or insertion methods? PMID- 21628026 TI - Isolated terminal ileal ulcer in asymptomatic patients: a call for prospective studies with long-term follow-up to determine the need for surveillance colonoscopy. PMID- 21628028 TI - Portal hypertensive polypoid enteropathy. PMID- 21628029 TI - Is the quality of endoscopic research improving? A 20-year review. PMID- 21628030 TI - "Dying with dignity". PMID- 21628031 TI - Palliative Care. Preface. PMID- 21628032 TI - Palliative care: an evolving field in medicine. AB - Palliative care is an approach that improves the quality of life of patients and their families facing the problems associated with life-threatening illness, through the prevention and relief of suffering by means of early identification and impeccable assessment and treatment of pain and other problems: physical, psychosocial, and spiritual. This article discusses illness trajectories and prognostic estimates, prognostic tools, educating physicians and nurses in palliative care, research in palliative medicine, and palliative care in hospitals and the community. PMID- 21628033 TI - Hospice care in the United States. AB - Hospice affirms the concept of palliative care as an intensive program that enhances comfort and promotes quality of life for individuals and their families. When cure is no longer possible, hospice recognizes that a peaceful and comfortable death is an essential goal of health care. Hospice believes that death is an integral part of the life cycle and that intensive palliative care focuses on pain relief, comfort, and enhanced quality of life for the terminally ill. Hospice also recognizes the potential for growth that exists within the dying experience for individuals and their families and seeks to protect and nurture this potential. PMID- 21628034 TI - Ethical issues in palliative care. AB - Ethical problems in medicine are common, especially when caring for patients at the end of life. However, many of these issues are not adequately identified in the outpatient setting. Primary care providers are in a unique and privileged position to identify ethical issues, prevent future conflicts, and help patients make medical decisions that are consistent with their individual values and preferences. This article describes some of the more common ethical issues faced by primary care physicians caring for patients with life-limiting illness. PMID- 21628035 TI - Assessment and management of pain in the terminally ill. AB - Regular assessment for the presence of pain and response to pain management strategies should be high priority in terminally ill patients. Pain management interventions are most effective when treatments are individualized based on the various physical and nonphysical components of pain at the end of life, and patients and family are educated and involved in the decision making. Opioids remain the cornerstone of pain management, and adjuvant analgesics and nonpharmacologic options are usually considered after relative stabilization of pain. This article describes the various issues that are pertinent to the assessment and treatment of pain in terminally ill patients. PMID- 21628036 TI - Assessment and management of gastrointestinal symptoms in advanced illness. AB - Primary care clinicians increasingly encounter patients with advanced illness, many suffering from symptoms other than pain. Key principles that guide palliative care must be incorporated into a plan of care for each patient and family. Although medical management continues to be the mainstay of treatment, the generalist in palliative care needs to be familiar with the patient's preferences and goals of care. This article provides an overview of gastrointestinal symptoms including anorexia, cachexia, nausea, vomiting, and constipation. Advanced progressive illnesses are defined here as incurable conditions that have significant morbidity in the later stages of illness. PMID- 21628037 TI - Management of end-stage dementia. AB - Dementia is a progressive and noncurable illness, and its management in late stages should follow a palliative care approach. However, many patients with advanced dementia sustain aggressive interventions that do not improve their survival and might hinder their comfort and quality of life. This is likely explained by a lack of research on this population; a lack of knowledge from health care providers, patients, and family members; and lack of communication between those caring for these patients. There is therefore an urgent need for research and education on this topic, as well as palliative care services devoted to this population. PMID- 21628038 TI - Management of end-stage heart failure. AB - The prevalence of heart failure (HF) is increasing and morbidity and mortality remain high. There is a clear need for palliative care for the growing population of chronically ill patients with HF. Because HF-specific therapy modifies disease and palliates symptoms, recommended treatments for chronic and acute decompensated HF are reviewed. This article discusses symptom burden in advanced HF and specific considerations for patients with HF regarding advance care planning and symptom-directed therapy. Options for care at the end of life, including hospice, chronic inotropic support, and deactivation of an internal cardiac defibrillator, are also discussed. PMID- 21628039 TI - Management of patients with end-stage chronic obstructive pulmonary disease. AB - Patients with end-stage chronic obstructive pulmonary disease (COPD) have poor quality of life, with limited activity, breathlessness, dependence on others, and recurrent needs for medical evaluation and treatment. Such patients demonstrate significant and progressive impairments in physical, mental, and social functioning. Because the rate of decline is variable, however, it is difficult to predict prognosis of survival. Currently available treatments only partially relieve symptoms, and patients become increasingly more disabled. This article reviews quality of life issues, proposed prognostic indicators, and pharmacologic and nonpharmacologic treatments in advanced COPD. Palliative measures to address breathlessness and unmet needs among patients with end-stage COPD are discussed. PMID- 21628040 TI - Palliative care in the treatment of end-stage renal failure. AB - Palliative care begins with establishing goals of care based on estimated prognosis in end-stage renal disease (ESRD). Patients with ESRD are increasingly characterized by older age and multiple comorbid illnesses, and have a mortality rate 8 times higher than the general Medicare population. Dialysis patients are appropriate for palliative care because of their high mortality rate and high symptom burden. More patients and families are choosing not to start or withdraw dialysis for multiple reasons, particularly in patients older than 60 years. Advance directives and resuscitation directives are important in ensuring compassionate and goal-directed palliative care of ESRD patients. Drug toxicities are avoidable by using appropriate drugs at the correct doses and dosing intervals. PMID- 21628041 TI - Palliative care in the management of advanced HIV/AIDS. AB - The basic elements of palliative care can be translated into practice for patients with HIV/AIDS. More than half of clinical events and deaths occurring among patients on highly active antiretroviral therapy are classified as non-AIDS illnesses. Thus, end-of-life care for patients with late-stage AIDS needs to include any palliative measures that are used for patients without AIDS. This article reviews the epidemiology of HIV/AIDS, prognostic indicators, opportunistic infections, specific AIDS-defining and non-AIDS-defining malignancies, substance abuse/liver disease, and highly active antiretroviral therapy and comfort measures for late-stage AIDS patients. PMID- 21628042 TI - Pediatric palliative care. AB - Progress in pediatric palliative care has gained momentum, but there remain significant barriers to the appropriate provision of palliative care to ill and dying children, including the lack of properly trained health care professionals, resources to finance such care, and scientific research, as well as a continued cultural denial of death in children. This article reviews the epidemiology of pediatric palliative care, special communication concerns, decision making, ethical and legal considerations, symptom assessment and management, psychosocial issues, provision of care across settings, end-of-life care, and bereavement. Educational and supportive resources for health care practitioners and families, respectively, are included. PMID- 21628043 TI - The Ulster hook for intermaxillary fixation. PMID- 21628044 TI - Global health, global health education, and infectious disease: the new millennium, part I. PMID- 21628045 TI - Global health: the twenty-first century global health priority agenda. AB - Growth in global health interest in the past 20 years has been overwhelming and many universities throughout the world have created departments or institutes of global health. The essence of global health has to be promoting health equity globally. The global health agenda must embrace design of mixed health systems, involving both private and public components to address the emerging threat of noncommunicable diseases and existing communicable diseases as well as to reduce health inequity. The priority agenda for the twenty-first century is challenging but the improvements of the past give hope that the barriers to improving global health are surmountable. PMID- 21628046 TI - Global health: evolving meanings. AB - Approaches to health, health care, and the terminology to describe global health have evolved over the past 70 years since the introduction of the Constitution of the World Health Organization and definition of health in broader terms. The early focus on individual care gradually shifted to community, population, and global approaches, with associated changes in the site of medical care, the personnel who provide it, and the education and training of those personnel. Concomitantly, goals changed from purely curative care to disease prevention and health promotion. Health was better understood to exist within the larger political, social, cultural, and ethical settings. PMID- 21628047 TI - Global health and the role of universities in the twenty-first century. AB - A vast gap exists between knowledge, generation of knowledge, and the application of knowledge to the needs and benefit of the global population. In middle-income and lower-income countries, universities are becoming more engaged with the communities in which they are located to try to solve the difficult problems of poverty and poor health. Global collaborations and reform of medical education in the twenty-first century will help move universities out of cloistered academic settings and into the community to bring the changes needed to equitably meet the health needs of all. PMID- 21628048 TI - Global health education consortium: 20 years of leadership in global health and global health education. AB - The Global Health Education Consortium (GHEC) is a group of universities and institutions committed to improving the health and human rights of underserved populations worldwide through improved education and training of the global health workforce. In the early 1990s, GHEC brought together many of the global health programs in North America to improve competencies and curricula in global health as well as to involve member institutions in health policy, development issues, and delivery of care in the inner cities, marginalized areas, and abroad. PMID- 21628049 TI - Global health: networking innovative academic institutions. AB - Medically underserved communities suffer a high burden of morbidity and mortality, increasing with remoteness where access to health services is limited. Major challenges are the overall shortage and maldistribution of the health workforce. There is a lack of understanding of how academic institutions can best contribute to addressing these health inequities. A new international collaborative of health professions schools, Training for Health Equity Network, is developing and disseminating evidence, challenging assumptions, and developing tools that support health profession institutions striving to meet the health and health workforce needs of underserved communities. PMID- 21628050 TI - Teaching the basics: core competencies in global health. AB - Compelling moral, ethical, professional, pedagogical, and economic imperatives support the integration of global health topics within medical school curriculum. Although the process of integrating global health into medical education is well underway at some medical schools, there remain substantial challenges to initiating global health training in others. As global health is a new field, faculties and schools may benefit from resources and guidance to develop global health modules and teaching materials. This article describes the Core Competencies project undertaken by the Global Health Education Consortium and the Association of Faculties of Medicine of Canada's Global Health Resource Group. PMID- 21628051 TI - Global health capacity and workforce development: turning the world upside down. AB - This article explores global health and the way in which the whole world is increasingly interdependent in terms of health. High-income countries need to help redress the balance of power and resources around the world, for self interest and self preservation if for no other reason. These countries have a particular responsibility to help support the training of more health workers and to strengthen health systems in low-income and middle-income countries. In this interdependent world, high-income countries can learn a great deal from poorer ones as well as vice versa, and concepts of mutuality and codevelopment will become increasingly important. PMID- 21628052 TI - The Infectious Diseases Institute at Makerere University, Kampala, Uganda. AB - The Infectious Diseases Institute (IDI) at Makerere University, Kampala, Uganda, was created in 2001. This article outlines its origins, principles, clinical programs, training activities, research programs, organizational structure, leadership, and contributions to Makerere University and its College of Health Sciences. PMID- 21628053 TI - Building a global health education network for clinical care and research. The benefits and challenges of distance learning tools. Lessons learned from the Hopkins Center for Clinical Global Health Education. AB - Expanding the capacity for clinical care and health research is a global priority and a global challenge. The Johns Hopkins Center for Clinical Global Health Education (CCGHE) was established in 2005 to provide access to high-quality training to health care providers in resource-limited settings. The CCGHE made a strategic decision to develop, use, and evaluate distance learning platforms to achieve its mission. In the initial years of this new program, several lessons have been learned that may be helpful to other programs considering the use of distance learning programs to expand global health clinical and research capacity. PMID- 21628054 TI - The Afya Bora Consortium: an Africa-US partnership to train leaders in global health. AB - The Afya Bora Consortium is a partnership of 8 academic health institutions, 4 in Africa and 4 in the United States. The Consortium is developing a Global Health Leadership Fellowship for medical, nursing, and public health professionals, largely drawn from the 4 African partner countries. The fellowship provides trainees with practical skills to prepare them for future positions leading the design, implementation, and evaluation of large, high-impact programs in governmental agencies, nongovernmental organizations, and academic health institutions in their own countries. This article describes a Pilot of the proposed program. PMID- 21628055 TI - Medical education: meeting the challenge of implementing primary health care in sub-Saharan Africa. AB - Medical education in sub-Saharan Africa must be defined by its health needs and the health care services required. The sociodemographic milieu that determines the disease pattern makes a compelling case for primary health care in the context of community participation and multisector community development as the driver of a plan for medical education, in tandem with clinicians' role in continuity of care. Such ideas have been derived from the experience of planning for undergraduate medical education at the Aga Khan University Medical College, Nairobi, whose curriculum incorporates broad-based general education and liberal arts principles. PMID- 21628056 TI - Health of migrants: working towards a better future. AB - For international migrants, in particular for those who cross without the required documentation, there are a number of health threats and problems that may begin during transit and a number that occur as a result of migrants' socio economic status in the receiving country. This article discusses the health status of Mexican immigrants in the United States including their access to health care, health disparities, and the social determinants of health among this population, with a focus on the health of women and children. PMID- 21628057 TI - Global health: governance and policy development. AB - Global health policy is now being influenced by an ever-increasing number of nonstate and non-intergovernmental actors to include influential foundations, multinational corporations, multi-sectoral partnerships, and civil society organizations. This article reviews how globalization is a key driver for the ongoing evolution of global health governance. It describes the massive increases in bilateral and multilateral investments in global health and it highlights the current global and US architecture for performing global health programs. The article closes describing some of the challenges and prospects that characterize global health governance today. PMID- 21628058 TI - The role of treaties, agreements, conventions, and other international instruments in global health. AB - As nations become more reliant on each other for cohesive development of global health policies and practice, and globalization increasingly makes health challenges in one part of the world concerns for all nations, the importance and use of international agreements in framing policy and national commitments have increased. This article reviews international agreements, looking specifically at multilateral instruments or partnerships, to identify those that either directly focus on or encompass health. It defines the different types of agreements, describes the process through which governments enter into these agreements, evaluates the legality of agreements under international law, and assesses participation by member states. PMID- 21628059 TI - [SOFMER and partnerships]. PMID- 21628060 TI - Overexpressing autoimmune regulator regulates the expression of toll-like receptors by interacting with their promoters in RAW264.7 cells. AB - Autoimmune regulator (Aire) is a transcriptional activator that regulates the ectopic expression of many tissue-restricted antigens in medullary thymic epithelial cells, and that has an important role in the negative selection of autoreactive T cells. However, the roles of Aire expression in peripheral lymphoid tissues and hematopoietic cells, especially monocytes/macrophages, remain poorly understood. In this study, we found that the mRNA and protein expression levels of toll-like receptor (TLR)1, TLR3, and TLR8 were notably up regulated in a mouse macrophage-like cell line (RAW264.7) stably expressing Aire, while the expression of TLR2, TLR4, TLR5, TLR6, TLR7, and TLR9 were not significantly changed. In addition, the mRNA expression of TLR3 and TLR8 were significantly increased in primary peritoneal macrophages transiently transfected with Aire. Using chromatin immunoprecipitation and a luciferase activity assay, we also found that Aire interacted with the TLR1, TLR3, and TLR8 promoters and increased the luciferase transcriptional activity of these promoters in RAW264.7 cells. Moreover, after stimulation by Pam(3)CSK(4), a TLR1 ligand, and poly(I:C), a TLR3 ligand, we found that the mRNA expression levels of IL-1alpha, TNFalpha, iNOS, and IFNalpha were increased in RAW264.7 cells stably expressing Aire. Together, these data suggest that Aire has a crucial role in the recognition of pathogenic microorganisms and peripheral immune tolerance in antigen-presenting cells (APCs) by regulating the expression of TLRs. PMID- 21628061 TI - Evaluation of different operation modes of high performance liquid chromatography for the analysis of complex mixtures of neutral oligosaccharides. AB - Chromatographic methods based on different HPLC operation modes, reverse phase (RP), high performance anion exchange chromatography (HPAEC), graphitized carbon chromatography (GCC) and hydrophilic interaction liquid chromatography (HILIC), have been developed and compared for the analysis of complex mixtures of neutral oligosaccharides with functional properties. Whereas GCC gave the best chromatographic separation of isomeric oligosaccharides with the same molecular weight (R(s) values in the range 1.0-4.0 and 2.4-5.6 for tetra- and pentasaccharides, respectively), HILIC provided the best results for mixtures including oligosaccharides of different degrees of polymerization (R(s) values of maltooligosaccharides between 3.4 and 6.2). Validation of the HILIC LC-MS method proved its utility for the analysis of oligosaccharide mixtures with functional properties: relative standard deviations lower than 10%, LOD's and LOQ's in the range 12.7-130.2 ng mL(-1) and 39.3-402.2 ng mL(-1), respectively, and linearity up to 10-20 MUg mL(-1). Quantitative data for fructooligosaccharides, gentiooligosaccharides and dextransucrase cellobiose acceptor oligosaccharides were obtained by using this method. PMID- 21628062 TI - Characterization of a 2.6 MUm Kinetex porous shell hydrophilic interaction liquid chromatography column in supercritical fluid chromatography with a comparison to 3 MUm totally porous silica. AB - The first systematic study of the performance of a porous shell, hydrophylic interaction liquid chromatography (HILIC) column in supercritical fluid chromatography (SFC) is presented. Observed efficiency on 2.6-MUm porous shell particles exceeded all reports using UHPLC on 100-mm long columns packed with <2 MUm totally porous particles. A Kinetex 4.6*150 mm, 2.6 MUm HILIC column significantly outperformed a 3 MUm Luna totally porous silica of the same length and diameter. A 17 component, low molecular weight test mix, consisting of a range of small drug-like molecules was separated isocratically on each column, with similar selectivity, but the porous shell column required 1/2 the time (~2 min vs. 4 min), with almost 50% higher efficiency. Even little retained compounds (k<0.5) exhibited more than 30,000 plates under some conditions. Reduced plate heights were higher than previously reported on porous shell particles in both HILIC and rHPLC, with the lowest value of 1.62. Significant fronting was sometimes observed. The cause of the fronting was not determined. The least symmetrical peaks showed the highest apparent efficiency. Pressure drop at optimum velocity (2.5 ml/min) and low modifier concentrations was <60 bar, and only exceeded 250 bar at near double optimum flow and 65% modifier. Peak widths were mostly just over 0.01 min (20 Hz) wide. There was a loss of efficiency when the injection volume was increased. The chromatograph was shown to have extremely low extra-column dispersion, on the order of 5-10 MUL(2), which is also the lowest reported in an SFC, in spite of using standard components. This is likely due to turbulent flow in the tubing and fittings. PMID- 21628063 TI - Experimental investigation of the difference in B-term dominated band broadening between fully porous and porous-shell particles for liquid chromatography using the Effective Medium Theory. AB - The difference in B-term diffusion between fully porous and porous-shell particles is investigated using the physically sound diffusion equations originating from the Effective Medium Theory (EMT). Experimental data of the B term diffusion obtained via peak parking measurements on six different commercial particle types have been analyzed (3 porous and 3 non porous). All particles were investigated using the same experimental design and test analytes, over a very broad range of retention factor values. First, the B-term reducing effect of the solid core (inducing an additional obstruction compared to fully porous particles) has been quantified using the Hashin-Shtrikman expression, showing that the presence of a solid core can account for a reduction of about 11% when the core diameter makes up 63% of the total particle diameter (Halo and Poroshell particles) and a reduction of 16% when the core diameter makes up 73% (Kinetex). Remaining differences can be attributed to differences in the microscopic structure of the meso-porous material (meso-pore diameter, internal porosity or relative void volume). The much lower B-term diffusion of Halo and Kinetex particles compared to the fully porous Acquity particles (some 20-40% difference, of which about 10-15% can be attributed to the presence of the solid core) can hence largely be attributed to the much smaller internal porosity and the smaller pore size of the meso-porous material making up the shell of these particles. PMID- 21628064 TI - Synthesis and characterization of silver nanoparticle and graphene oxide nanosheet composites as a bactericidal agent for water disinfection. AB - Graphene oxide (GO) nanosheets impregnated with silver nanoparticles (Ag NPs) were fabricated by the in situ reduction of adsorbed Ag(+) by hydroquinone (HQ) in a citrate buffer solution. Paper-like Ag NP/GO composite materials were fabricated owing to convenient structure characterization and antibacterial tests. The Ag NP/GO composites were characterized by UV-vis spectra, transmission electron microscope, electron diffraction, Raman spectroscopy, and field emission scanning electron microscope coupled with Energy Dispersive Spectrometer. Antibacterial activity was tested using Escherichia coli and Staphylococcus aureus as model strains of Gram negative and Gram positive bacteria, respectively. The as-prepared composites exhibit stronger antibacterial activity against both. The Ag NP/GO composites performed efficiently in bringing down the count of E. coli from 10(6) cfu/mL to zero with 45 mg/L GO in water. The micron scale GO nanosheets (lateral size) enable them to be easily deposited on porous ceramic membranes during water filtration; making them a promising biocidal material for water disinfection. PMID- 21628065 TI - Gait disturbances in Parkinson disease. Did freezing of gait exist before levodopa? Historical review. AB - Gait disturbances occur frequently in advanced Parkinson's disease (PD) including slow gait, postural changes, festination and freezing of gait. We have reviewed descriptions of gait abnormalities in PD from classic predopa-literature and compared them with those found in contemporary references. Several components of gait disturbances associated with shaking palsy were very well known in classic literature. James Parkinson, Charcot, Gowers and Wilson described slowness of gait, postural changes, loss of postural reflexes and festination; according to James Parkinson, festination was a pathognomonic element in shaking palsy. In contrast, freezing of gait was rarely mentioned in historic literature save for anecdotal reports (Buzzard 1888). Freezing of gait was fully noticed after the chronic use of levodopa (Barbeau and Ambani). In this historical review, we analyze the concept, identification and evolution of gait disturbances in PD through the time. PMID- 21628066 TI - Fate of perfluorooctanesulfonate and perfluorooctanoate in drinking water treatment processes. AB - Perfluorooctanesulfonate (PFOS) and perfluorooctanoate (PFOA) have been recognized as global environmental pollutants. Although PFOS and PFOA have been detected in tap water from Japan and several other countries, very few studies have examined the fate, especially removal, of perfluorinated compounds (PFCs) in drinking water treatment processes. In this study, we analyzed PFOS and PFOA at every stages of drinking water treatment processes in several water purification plants that employ advanced water treatment technologies. PFOS and PFOA concentrations did not vary considerably in raw water, sand filtered water, settled water, and ozonated water. Sand filtration and ozonation did not have an effect on the removal of PFOS and PFOA in drinking water. PFOS and PFOA were removed effectively by activated carbon that had been used for less than one year. However, activated carbon that had been used for a longer period of time (>1 year) was not effective in removing PFOS and PFOA from water. Variations in the removal ratios of PFOS and PFOA by activated carbon were found between summer and winter months. PMID- 21628067 TI - Degradation of phenol in mists by a non-thermal plasma reactor. AB - A link tooth wheel-cylinder non-thermal plasma reactor was set up to investigate the degradation of phenol in the mists. In addition, the decomposition efficiency of phenol, TOC removal, and byproduct formation were investigated. The stable discharge was achieved in both air and the mist condition. The decomposition efficiency and TOC removal increased with increasing the input power. For the input power of 3.6 W, the phenol decomposition and TOC removal reached 90% and 47%, respectively. Phenol degradation byproducts were identified as small molecular organic acids, including formic acid, acetic acid, and oxalic acid. Their masses in the trapped solutions first increased and then decreased slightly with increasing the input power. Therefore, the biodegradation capacity of the phenol degradation byproducts can be improved. PMID- 21628069 TI - TRP channels in cell survival and cell death in normal and transformed cells. AB - TRP channels form a superfamily of channel proteins exhibiting versatile regulatory characteristics with many channels participating in the regulation of Ca(2+) homeostasis and influencing the cell fate. Multitude of evidence is emerging that the colocalization of TRP channels with Ca(2+)-sensing elements of specific regulatory pathways leading to either proliferation or apoptosis is what makes these channels participate in cell fate regulation and, in turn, determines the final effect of Ca(2+) entry via the particular channel. This review focuses on the aspects of TRP channel localization and function that affect the balance between cell survival and death and how various dysregulations of these channels may lead to perturbed balance and onset of cancer. PMID- 21628071 TI - t(8;22)/BCR-FGFR1 myeloproliferative disorder presenting as B-acute lymphoblastic leukemia: report of a case treated with sorafenib and review of the literature. PMID- 21628070 TI - Bcl-2 interaction with the inositol 1,4,5-trisphosphate receptor: role in Ca(2+) signaling and disease. AB - The Bcl-2 protein, best known for its ability to inhibit apoptosis, interacts with the inositol 1,4,5-trisphosphate receptor (IP(3)R) Ca(2+) channel to regulate IP(3)-mediated Ca(2+) release from the endoplasmic reticulum. This review summarizes the current state of knowledge regarding the interaction of Bcl 2, and also its homologue Bcl-xl, with the IP(3)R and how these interactions regulate Ca(2+) signaling. The dual role of these interactions in promoting prosurvival Ca(2+) signals, while at the same time inhibiting proapoptotic Ca(2+) signals, is discussed. Moreover, this review will elucidate the recently recognized importance of the Bcl-2-IP(3)R interaction in human disease. PMID- 21628072 TI - A monoclonal antibody distinguishes between two IgM heavy chain isotypes in Atlantic salmon and brown trout: protein characterization, 3D modeling and epitope mapping. AB - Atlantic salmon (Salmo salar) and brown trout (Salmo trutta) possess two distinct subpopulations of IgM which can be separated by anion exchange chromatography. Accordingly, there are two isotypic MU genes in these species, related to ancestral tetraploidy. In the present work it was verified by mass spectrometry that IgM of peak 1 (subpopulation 1) have heavy chains previously designated as MUB type whereas IgM of peak 2 (subpopulation 2) have heavy chains of MUA type. Two adjacent cysteine residues are present near the C-terminal part of MUB, in contrast to one cysteine residue in MUA. Salmon IgM of both peak 1 and peak 2 contain light chains of the two most common isotypes: IgL1 and IgL3. In contrast to salmon and brown trout, IgM of rainbow trout (Oncorhynchus mykiss) is eluted in a single peak when subjected to anion exchange chromatography. Surprisingly, a monoclonal antibody MAb4C10 against rainbow trout IgM, reacted with MUA in salmon, whereas in brown trout it reacted with MUB. It is plausible to assume that DNA has been exchanged between the paralogous A and B loci during evolution while maintaining the two sub-variants, with and without the extra cysteine. MAb4C10 was conjugated to magnetic beads and used to separate cells, demonstrating that MU transcripts residing from captured cells were primarily of A type in salmon and B type in brown trout. An analysis of amino acid substitutions in MUA and MUB of salmon and brown trout indicated that the third constant domain is essential for MAb4C10 binding. This was supported by 3D modeling and was finally verified by studies of MAb4C10 reactivity with a series of recombinant MU3 constructs. PMID- 21628073 TI - Phosphatidylserine inhibits NFkappaB and p38 MAPK activation in human monocyte derived dendritic cells. AB - Phosphatidylserine (PS) is an anionic phospholipid restricted to the inner surface of the plasma membrane. PS translocates to the cell surface during early apoptosis where it serves as a marker for rapid uptake by phagocytes. PS is also thought to regulate immune responses. Dendritic cells (DC) are the most potent antigen presenting cells. Previous studies demonstrated that PS inhibits the expression of MHC and co-stimulatory molecules, the secretion of IL-12p70, and the ability to activate T cells by human monocyte derived DCs. However, the cell signaling mechanisms by which PS regulated DCs are not well described. In the current study we tested the effects of PS on signal transduction pathways thought to regulate human myeloid DC maturation and IL-12p70 production. We showed that PS inhibited the activation of nuclear factor-kappaB (NFkappaB) in response to LPS by preventing IkappaBalpha phosphorylation and degradation. PS also increased the total IkappaBalpha levels in immature DCs and inhibited p38 mitogen activated protein kinase (MAPK) phosphorylation and activation. The findings suggest a possible mechanism for regulating the immunostimulatory function of DCs by PS. PMID- 21628074 TI - Fish consumption and severely depressed mood, findings from the first national nutrition follow-up study. AB - The evidence obtained from prospective studies to support the hypothesis that fish consumption may improve mental status remains limited. The current study prospectively assessed a low frequency of fish consumption as a risk factor for depressed mood. Included were 5068 adults aged 25-74 years examined in 1971-1975 as the baseline of the First National Health and Nutrition Examination Survey Follow-up Study. Frequency of eating fish at baseline was obtained using a 3 month food frequency questionnaire. Severely depressed mood (SDM) was defined as the Center for Epidemiologic Studies Depression Scale scores >=22 or taking anti depressants. After an average of 10.6 years of follow-up, among men (n=2039), the percentage of individuals with SDM was 11.7%. Compared with frequent consumers (more than once a week), the odds ratios (ORs) were 1.43 (95%CI=0.66-3.11) and 2.08 (1.08-4.09) respectively for the men eating fish once a week and less than once a week (p for trend=0.03). Among women (n=3029), the percentage of individuals with SDM was 17.89%. The ORs were 1 (reference), 0.91 (0.68-1.22) and 1.15 (0.83-1.59) respectively for the women eating fish more than once, once, and less than once a week. These estimates were obtained after adjustment for indicators of social deprivation and major physical diseases. The study concluded that independently from social deprivation and physical diseases, low fish consumption was a risk factor for SDM among men. Further studies are needed to confirm these findings and elucidate mechanisms for the difference between men and women. PMID- 21628075 TI - beta-Glucan plus ascorbic acid in neonatal calves modulates immune functions with and without Salmonella enterica serovar Dublin. AB - To determine if beta-glucan plus ascorbic acid affects adherence and pathogenicity of Salmonella Dublin and innate immune response in neonatal calves, 20 calves were fed control or supplemented diets (beta-glucan, 0.9 g/d, plus ascorbic acid, 500 mg/d) until d 23. On d 21, 5 calves per treatment received 2.4 * 10(8)CFU of S. Dublin orally. S. Dublin spread through intestinal tissues into mesenteric lymph nodes (MLN), spleen, and lung tissues within 48 h. All supplemented calves had less mRNA expression of IL-1 receptor antagonist in liver. Leukocyte cell surface markers changed in lung cells, but not in blood, MLN, or spleen. CD14 in lungs was greatest for calves receiving supplement and challenge, but CD18 in lungs was greater for challenged than control calves. Lung DEC205 was greatest for challenged calves with and without supplement compared to controls, but more lung cells expressed CD14 for all treated groups compared to controls. These data show that S. Dublin briefly inhabited the intestinal tract, moving quickly to spleen, MLN, and lung tissues. Lung tissue was modulated by S. Dublin, but supplement alone increased CD14 expressing cells. The supplement appears not to attenuate invasiness but modified some lung cell populations by 48h. PMID- 21628076 TI - Morbid RANKIN score strongly predicts in-patient mortality within 90 days in older people in both acute and community care settings independently of FANGG (fracture, acquired neurological deficit or any geriatric giants) and acute illness markers (AIMs). AB - Understanding the relative impact of factors that are associated with poor outcome of older people admitted either to an acute or a rehabilitation setting is essential to further our knowledge in provision of appropriate care. We conducted a prospective study to examine whether FANGG and AIMs are important prognostic indicators of mortality outcome in hospitalised older people when patients' morbid functional status is considered. Participants were two consecutive series of 200 patients admitted to care of the elderly wards in an acute teaching hospital and a community hospital in Birmingham, UK during April to August 2004 and the same months in 2005. The association with the outcome of mortality was examined in a univariate analysis, and then multiple logistic regression models. A total of 400 patients (men 116, 29.2%) were included in this study (mean age=85.3 years, range 64-104 years). There were 72 in-hospital deaths (18.0%). The prevalence of FANGG is low in this series; the majority (89.3%) had none or only one factor. The adjusted analysis showed that only age (p=0.05), gender (p=0.01) and morbid Rankin score (p<0.001) were predictive of outcome. The morbid Rankin score of 5 had over threefold higher likelihood of dying as in patient (OR=3.31; 95% CI=1.48-7.40, p=0.003) independently of age, gender, site, FANGG, Rankin*AIMs interaction and AIMs. The morbid Rankin score strongly predicts the in-patient mortality over and above of age, gender, FANGG and AIMs in this patient population. PMID- 21628077 TI - Effect of subjective knee-joint pain on the laterality of knee extension strength and gait in elderly women. AB - This study aimed to examine the effect of subjective knee-joint pain on the laterality of knee extension strength and gait in elderly women. The subjects were 144 elderly women (62-94 years old; mean age 76.2+/-6.0 years; +/-S.D.) who were divided into the following groups: 81 persons without knee-pain (no knee pain group), 39 persons with the subjective pain in right or left knee (single knee-pain group), and 24 persons with the subjective pain in both knees (double knee-pain group). The subjects took a knee extension strength test and a 12 m maximum effort walk test. Knee extension strength, stance time, swing time, stride length, step length and swing speed were selected as parameters. A significant laterality was found in knee extension strength only in the one knee pain group. The laterality of gait parameters was not found in all groups. In conclusion, elderly women who can perform daily living activity independently, even though having subjective pain in either knee or laterality in knee extension strength exertion show little laterality of gait during short distance walking. PMID- 21628078 TI - Which body mass index (BMI) is better in the elderly for functional status? AB - BMI is commonly used indicator of malnutrition and 18.5-24.9 kg/m(2) is generally regarded optimal. However, there is an ongoing debate on ideal range for elderly. BMI cut-off values vary also between ethnic groups. We aimed to investigate relationships between BMI, functional status and malnutrition in elderly living in a nursing home in Turkey. BMIs of 254 residents were calculated. Chronic diseases and currently used drugs were noted. Functional status was evaluated with Katz-activities-of-daily-living (ADL) and Lawton-instrumental-activities-of daily-living (IADL). Nutritional assessment was performed by Mini-Nutritional Assessment (MNA) test. Mean age was 75.2 +/- 8.2 years. Subjects were classified into 4 groups as BMI <18.5, 18.5-24.9, 25-29.9, and >= 30.0 kg/m(2). ADL scores and IADL scores were higher in higher BMI groups. There were no differences in terms of age-number of chronic diseases. Even in BMI >= 35 kg/m(2) residents, ADL was significantly higher than 25-34.9 kg/m(2) residents. BMI was significantly correlated with ADL and IADL scores. In Groups 3 and 4, there were 22.2% and 9.1% residents without normal nutrition, respectively. Better functional status was associated with higher BMI values even in BMIs >= 30 kg/m(2). In elderly, relative high rates of undernutrition may be present in BMIs regarded as overweight or obese. PMID- 21628079 TI - Physicians' participation in the Physicians' Quality Reporting Initiative and their perceptions of its impact on quality of care. AB - BACKGROUND: Medicare established the Physicians Quality Reporting Initiative (PQRI, recently renamed the Physicians Quality Reporting System) to increase reporting of quality metrics and promote healthcare quality. OBJECTIVE: To identify characteristics of PQRI participants and examine their beliefs about its impact. DESIGN: National survey of 4934 U.S. physicians, conducted June through October 2009. SETTING: All practice settings. PARTICIPANTS: Randomly selected physicians categorized as primary care, medical specialists, surgeons, other specialists. MEASUREMENTS: Beliefs about impact of PQRI reporting on quality. RESULTS: The response rate was 49.8%. There were no significant differences between respondents and non-respondents by age, gender, specialty, and region. Thirty-eight percent participated in the PQRI, and were more likely than non participants to be practice owners (69.0% vs. 57.1%, p<.0001) and to receive performance bonuses through their employer or practice (50.4% vs. 37.0%, p<.0001). Half of PQRI participants believed it had no impact on quality. Medical specialists (57.0%) and surgeons (55.1%) were more likely than primary care (40.4%) and other physicians (45.7%) to say that PQRI has no impact on quality (p=.004). CONCLUSIONS: Most PQRI participants believed it had little if any impact on quality. Medicare should identify the reasons behind physicians' negative views while it works to expand the Physicians Quality Reporting System. PMID- 21628080 TI - Milk and sugar: regulation of cell wall synthesis in the milk yeast Kluyveromyces lactis. AB - The milk yeast Kluyveromyces lactis is an alternative model yeast to the well established Saccharomyces cerevisiae. The cell wall of these fungi consists of polysaccharides (i.e. long chains of beta-1,3- and beta-1,6-linked sugar chains and some chitin) and mannoproteins, both of which are continually adapted to environmental conditions in terms of their abundance and organization. This implies the need to perceive signals at the cell surface and to transform them into a proper cellular response. The signal transduction cascade involved in this process is generally referred to as the cell wall integrity (CWI) pathway. CWI signaling and cell wall composition have been extensively studied in the Baker's yeast S. cerevisiae and are also of interest in other yeast species with commercial potential, such as K. lactis. We here summarize the results obtained in the past years on CWI signaling in K. lactis and use a comparative approach to the findings obtained in S. cerevisiae to highlight special adaptations to their natural environments. PMID- 21628081 TI - Could LogP be a principal determinant of biological activity in 18-crown-6 ethers? Synthesis of biologically active adamantane-substituted diaza-crowns. AB - 18-crown-6 ethers are known to exert their biological activity by transporting K(+) ions across cell membranes. Using non-linear Support Vector Machines regression, we searched for structural features that influence antiproliferative activity in a diverse set of 19 known oxa-, monoaza- and diaza-18-crown-6 ethers. Here, we show that the logP of the molecule is the most important molecular descriptor, among ~1300 tested descriptors, in determining biological potency (R(2)(cv) = 0.704). The optimal logP was at 5.5 (Ghose-Crippen ALOGP estimate) while both higher and lower values were detrimental to biological potency. After controlling for logP, we found that the antiproliferative activity of the molecule was generally not affected by side chain length, molecular symmetry, or presence of side chain amide links. To validate this QSAR model, we synthesized six novel, highly lipophilic diaza-18-crown-6 derivatives with adamantane moieties attached to the side arms. These compounds have near-optimal logP values and consequently exhibit strong growth inhibition in various human cancer cell lines and a bacterial system. The bioactivities of different diaza-18-crown-6 analogs in Bacillus subtilis and cancer cells were correlated, suggesting conserved molecular features may be mediating the cytotoxic response. We conclude that relying primarily on the logP is a sensible strategy in preparing future 18 crown-6 analogs with optimized biological activity. PMID- 21628082 TI - Association between population density and reported incidence, characteristics and outcome after out-of-hospital cardiac arrest in Sweden. AB - AIM: To describe the reported incidence of out of hospital cardiac arrest (OHCA) and the characteristics and outcome after OHCA in relation to population density in Sweden. METHODS: All patients participating in the Swedish Cardiac Arrest Register between 2008 and 2009 in (a) 20 of 21 regions (n=6457) and in (b) 165 of 292 municipalities (n=3522) in Sweden, took part in the survey. RESULTS: The regional population density varied between 3 and 310 inhabitants per km(2) in 2009. In 2008-2009, the number of reported cardiac arrests varied between 13 and 52 per 100,000 inhabitants and year. Survival to 1 month varied between 2% and 14% during the same period in different regions. With regard to population density, based on municipalities, bystander CPR (p=0.04) as well as cardiac etiology (p=0.002) were more frequent in less populated areas. Ambulance response time was longer in less populated areas (p<0.0001). There was no significant association between population density and survival to 1 month after OHCA or incidence (adjusted for age and gender) of OHCA. CONCLUSION: There was no significant association between population density and survival to 1 month after OHCA or incidence (adjusted for age and gender) of OHCA. However, bystander CPR, cardiac etiology and longer response times were more frequent in less populated areas. PMID- 21628083 TI - Speciation of selenium in groundwater: seasonal variations and redox transformations. AB - Speciation of selenium in groundwater is essential from the viewpoint of toxicity to organisms and biogeochemical cycling. Selenium speciation in groundwater is controlled by aquifer redox conditions, microbial transformations, dissolved oxygen (DO) and other redox couples. A suburban area of Chennai city in India, where improper waste disposal measures have been practiced is selected for this study. Se(IV), Se(VI) and other hydrochemical parameters were monitored in shallow ground water during pre- and post-monsoon seasons for a period of three years. The objective of the study was to investigate the effect of groundwater recharge on selenium speciation. The concentration of Se(IV), and Se(VI) ranged between 0.15-0.43 MUg L(-1) and 0.16-4.73 MUg L(-1), respectively. During post monsoon period the concentration of Se(IV), and Se(VI) ranged between 0.15-1.25 MUg L(-1) and 0.58-10.37 MUg L(-1), respectively. Se(VI) was the dominant species of selenium during the pre- and post-monsoon periods. During the post-monsoon periods, leaching of selenium from soil was more effective due to the increased oxidizing nature of the groundwater as indicated by the DO and redox potential (Eh) measurements. This finding has important implications on the behavior of selenium in groundwater, and also on the health of people consuming groundwater from seleniferous areas. PMID- 21628084 TI - Botulinum toxin injection for management of thoracic outlet syndrome: a double blind, randomized, controlled trial. AB - We studied the effect of botulinum toxin type A (BTX-A) injections to the scalene muscles on pain in subjects with thoracic outlet syndrome (TOS) in this double blind, randomized, parallel group trial with follow-up at 6 weeks, 3 months, and 6 months. Thirty-eight patients referred to physiatrists for management of TOS with BTX-A injection were included. One subject was lost to follow-up and all other subjects completed the trial. A 75-unit dose of BTX-A reconstituted with 0.75 cc of normal saline was injected to the anterior scalene (37.5 units) and middle scalene (37.5 units) muscles using electromyographic guidance. The primary outcome measure was pain as measured on a horizontal visual analog scale (VAS) 6 weeks-post-injection. Secondary outcomes were paresthesias measured on a VAS and function measured with the Disabilities of the Arm, Shoulder and Hand (DASH) and Short-form 36 (SF-36) questionnaires. For the primary outcome measure of VAS scores for pain at 6 weeks, the difference in the means adjusted for baseline VAS scores between placebo and BTX-A was 5.03 mm in favor of BTX-A (95% confidence interval -15.7 to 5.7, P=.36). Changes in secondary outcome measures were also not statistically significant. We conclude that BTX-A injections to the scalene muscles did not result in clinically or statistically significant improvements in pain, paresthesias, or function in this population of subjects with TOS. PMID- 21628085 TI - WITHDRAWN: Major proteins from the seminal plasma of adult Santa Ines rams. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 21628086 TI - Histopathologic correlation of hepatocellular carcinoma after transcatheter arterial chemoembolization with polyvinyl alcohol particle of various sizes. AB - AIM: Transcatheter arterial chemoembolization (TACE) is one of major treatment for unresectable hepatocellular carcinomas (HCC) and has been used as a neoadjuvant treatment before surgery. This study was to describe the histopathologic features of HCC after TACE with variously sized polyvinyl alcohol (PVA) particles. MATERIALS AND METHODS: Seventeen patients undergoing TACE with PVA followed by surgery for HCC were analyzed. The PVA particles used in TACE were categorized into two groups in respects of particle sizes: the group I, 47 90 MUm (n=8) and the group II, >90-250 MUm (n=9). The histopathologic features of the resected HCC were characterized with the emphasis on the number of thrombosed vessels and minimal diameter of arterioles/capillaries containing polyvinyl alcohol particles. The clinical results after TACE were also addressed. RESULTS: Histopathologic examinations showed that the median minimal diameters of arterioles containing PVA particles were 0.035 mm in group I and 0.06 mm in group II (p=0.0078). We observed the PVA particle in the sinusoidal spaces of non tumourous liver in only one patient. However, no sinusoidal infarction was demonstrated in either group. Mean tumour necrosis rate was 67% vs. 61% for the group I and II, respectively. CONCLUSIONS: The smaller PVA particles can reach and occlude more distal arteriolar capillaries, but rarely leak into non tumourous hepatic sinusoidal spaces. Slightly better tumour necrosis rate after TACE can be achieved. PMID- 21628087 TI - Application of a new optimization strategy for the separation of tertiary alkaloids extracted from Strychnos usambarensis leaves. AB - The HPLC separation of six alkaloids extracted from Strychnos usambarensis leaves has been developed and optimized by means of a powerful methodology for modelling chromatographic responses, based on three steps, i.e. design of experiments (DoE), independent component analysis (ICA) and design space (DS). This study was the first application of a new optimization strategy to a complex natural matrix. The compounds separated are the isomers isostrychnopentamine and strychnopentamine, 10-hydroxyusambarine and 11-hydroxyusambarine, also strychnophylline and strychnofoline. Three LC parameters have been optimized using a multifactorial design comprising 29 experiments that includes 2 center point replicates. The parameters were the percentage of organic modifiers used at the beginning of a gradient profile which consisted in different proportions of methanol (MeOH) and acetonitrile (MeCN), the gradient time to reach 70% of organic modifiers starting from the initial percentage and the percentage of MeCN found in the mobile phase. Subsequent to the experimental design application, predictive multilinear models were developed and used in order to provide optimal analytical conditions. The optimum assay conditions were: methanol/acetonitrile sodium pentane sulfonate (pH 2.2; 7.5 mM) (33.4:66.6, v/v) at a mobile phase flow rate of 1 ml/min during a 40.6 min gradient time. The initial organic phase contained 3.7% MeCN and 96.3% MeOH. The method showed good agreement between the experimental data and predictive value throughout the studied parameters space. Improvement of the analysis time and optimized separation for the compounds of interest was possible due to the original and powerful tools applied. Finally, this study permitted the acquisition of isomers profiles allowing the identification of the optimal collecting period of S. usambarensis. PMID- 21628088 TI - Characterization of drug-cyclodextrin formulations using Raman mapping and multivariate curve resolution. AB - Raman chemical imaging was used in the characterization of drug-excipient interactions between a drug and different types of cyclodextrins. Detailed analysis was carried out regarding the interactions between the active ingredient (API) and the cyclodextrins and the heterogeneity of the samples was studied using multivariate curve resolution-alternating least squares algorithm. The amount of recrystallized pure API was also estimated using the same curve resolution method. The Raman mapping results were validated via scanning electron microscopy-energy dispersive X-ray spectroscopy and X-ray powder diffraction. Raman mapping was found to be suitable to detect traces of pure crystalline API below the detection limit of X-ray powder diffraction. PMID- 21628089 TI - Treatment for breast sarcoma: a large, single-centre series. AB - AIMS: To report outcomes in breast sarcoma in the context of a major series from a tertiary referral centre. METHODS: Retrospective analysis was performed on patients with histologically-proven breast sarcoma treated between 1996 and 2006. Kaplan-Meier survival curves were constructed and differences assessed by Log Rank and Wilcoxon tests. RESULTS: 63 patients were identified; 57 underwent treatment with curative intent. 24 patients had undergone previous radiotherapy. 36 patients who underwent primary surgery elsewhere were referred for further treatment, of which 22 had at least one involved margin from primary resection. Surgery performed and margins status varied between patients undergoing primary surgery at this institution (n = 21; WLE = 8, mastectomy = 12, chest wall resection = 1, involved margins = 2 [10%]) or at a referring institution (n = 36; lumpectomy = 25, mastectomy = 11, involved margins = 22 [61%]), although there was no difference in tumour size or previous radiotherapy status. Previous irradiation was associated with poor prognosis. A greater proportion of these patients required primary mastectomy to ensure adequate clearance; the majority of the post-irradiation tumours were angiosarcomas (15/19) and significantly more relapsed locally (P < 0.001). All patient disease-free survival (DFS) rates were 71% at 2 and 42% at 5 years. DFS improved when primary surgery was undertaken at a high-volume sarcoma unit; 2-yr 84%vs75%; 5-yr 58%vs37%. There was a trend towards worse DFS with increasing size and increasing grade of tumour but this did not attain significance. CONCLUSIONS: Radiation-induced breast sarcoma has worse local recurrence rates compared to primary breast sarcoma. Involved margins were fewer at a specialist unit, which may translate into improved outcome. PMID- 21628090 TI - Prognosis of early-stage synchronous bilateral invasive breast cancer. AB - BACKGROUND: Contradictory data exists concerning the prognosis of patients with synchronous bilateral breast cancer (SBBC). Most authors report a worse prognosis for SBBC patients compared to unilateral breast cancer (UBC) patients. There are a few studies that did not support these findings. This study gives a comprehensive picture of SBBC and tests the hypothesis that outcome of this entity is based on the tumor with the worse prognosis (reference lesion). PATIENTS & METHODS: The data of two prospective Swiss breast cancer databases covering a 20-year period (1990-2009) was reviewed. Forty-six cases of SBBC were identified. In 34 patients with early-stage SBBC, the reference lesions (defined as the tumor with the more advanced stage or, in cases where both tumors had the same stage, the larger tumor) were compared in a case-control approach with 100 patients having UBC (SBBC/UBC ratio = 1/3). The controls were matched for age, time of diagnosis, tumor size, axillary node status, histological grade and estrogen-receptor status. Differences in terms of survival curves were analyzed using the log rank test; the possible correlation between matched groups was evaluated by a frailty Cox model. RESULTS: There were no significant differences in disease-specific survival between SBBC and its unilateral controls (HR, 0.932; 95% CI, 0.322-1.07; p = 0.90). CONCLUSIONS: The prognosis of SBBC was determined by the reference lesion; the contralateral second tumor had no additional impact on outcome. PMID- 21628091 TI - Language and verbal short-term memory skills in children with Down syndrome: a meta-analytic review. AB - This study presents a meta-analytic review of language and verbal short-term memory skills in children with Down syndrome. The study examines the profile of strengths and weaknesses in children with Down syndrome compared to typically developing children matched for nonverbal mental age. The findings show that children with Down syndrome have broad language deficits (that are not restricted to measures of expressive language) and associated verbal short-term memory deficits. The profile of language skills in children with Down syndrome shows similarities to that seen in children with Specific Language Impairment. The practical and theoretical implications of these findings are discussed. PMID- 21628092 TI - The influence of size, structure and hydrophilicity of model surfactants on the adsorption of lysozyme to oil-water interface--interfacial shear measurements. AB - The flexibility and aggregation of proteins can cause adsorption to oil-water interfaces and thereby create challenges during formulation and processing. Protein adsorption is a complex process and the presence of surfactants further complicates the system, in which additional parameters need to be considered. The purpose of this study is to scrutinize the influence of surfactants on protein adsorption to interfaces, using lysozyme as a model protein and sorbitan monooleate 80 (S80), polysorbate 80 (T80), polyethylene-block-poly(ethylene glycol) (PE-PEG) and polyglycerol polyricinoleate (PG-PR) as model surfactants. Rheological properties, measured using a TA AR-G2 rheometer equipped with a double wall ring (DWR) geometry, were used to compare the efficacy of the surfactant in hindering lysozyme adsorption. The system consists of a ring and a Delrin(r) trough with a circular channel (interfacial area=1882.6 mm(2)). Oscillatory shear measurements were conducted at a constant frequency of 0.1 Hz, a temperature of 25 degrees C, and with strain set to 1%. The adsorption of lysozyme to the oil-water interface results in the formation of a viscoelastic film. This can be prevented by addition of surfactants, in a manner depending on the concentration and the type of surfactant. The more hydrophilic surfactants are more effective in hindering lysozyme adsorption to oil-water interfaces. Additionally, the larger surfactants are more persistent in preventing film formation, whereas the smaller ones eventually give space for the lysozyme on the interface. The addition of a mixture of two different surfactants was only beneficial when the two hydrophilic surfactants were mixed, in which case a delay in the multilayer formation was detected. The method is able to detect the interfacial adsorption of lysozyme and thus the hindering of film formation by model surfactants. It can therefore aid in processing of any delivery systems for proteins in which the protein is introduced to oil-water interfaces. PMID- 21628093 TI - Cajanus indicus leaf protein: Beneficial role in experimental organ pathophysiology. A review. AB - The herb, Cajanus indicus L, has been and is popular for its medicinal value in India and other countries for long. The herb is mainly cultivated for the seeds which are used as pulses and are rich in proteins. People of rural India and some neighboring countries use the aqueous extract of the leaves of the herb against poor liver function and recently it has been found that the extract is not only useful against liver damage but also beneficial for renal failure and a number of other pathophysiological conditions. Intraperitoneal administration of the aqueous protein fraction of the leaves has shown hepatoprotective activity in mice. The protein fraction revealed the presence of a 43kDa protein having antioxidant and other protective properties in organ pathophysiology. The purified protein, CI-protein, scavenges free radicals generated by different free radical inducers and helps providing cytoprotection. Amino acid sequence of CI protein has some structural similarity with plastocyanin, an electron carrier protein in photosynthesis. The protein has also been found to be active against a number of organ dysfunction inducer chemicals and drugs, like carbon tetrachloride, thioacetamide and acetaminophen. Signal transduction studies suggest that CI-protein exerts its protective action by free radical scavenging and antioxidative properties; it activates NF-kappaB and Akt without any involvement of ERK1/ERK2 and STAT-3 in acetaminophen induced hepatic pathophysiology. Besides, it reduces both drug and toxin induced cytotoxicity by decreasing the formation and/or scavenging of free radicals involving cytochrome P450, taking part in detoxification of xenobiotics. PMID- 21628094 TI - Congruence between international guidelines and mite specific immunotherapy prescribing practices. AB - Both rhinitis (ARIA) and asthma (GINA) guidelines recommend allergen-specific immunotherapy (SIT) tailored to the specific levels of severity of each disease. Real world studies evaluating congruence between these recommendations and prescribing practice in the single patient with comorbidity are lacking. An observational polycentric study was carried out in 518 patients recruited from 34 allergy centers throughout Italy. A questionnaire was administered to each consecutive patient over a span of four months. Taking into account guideline recommendations for both diseases, concomitant in the same patient, three subsets resulted: patients not eligible for SIT (11%); patients eligible for SIT for one disease only (60%); patients eligible for SIT for both diseases (29%). SIT was prescribed in 257 (49.6%) subjects. The level of SIT prescription was about 50% in all three groups. Consistent with the ARIA guidelines, a correlation between the prescription of SIT and the severity of rhinitis was documented (r=0.87; p=0.001). An association with asthma severity was found (p=0.02), but the trend was inconsistent with the GINA recommendations. Young age was the most important factor for SIT prescription both in the eligible for one disease and in the eligible for both diseases subset. The tendency towards worsening of symptoms was a factor for SIT in the eligible for one disease subset. In mite allergic patients with rhinitis and asthma comorbidity, the severity of rhinitis and the young age are the most important factors driving the SIT prescription. The congruence of SIT prescription was better for the ARIA than GINA guidelines. PMID- 21628095 TI - Visual detection of Hg2+ in aqueous solution using gold nanoparticles and thymine rich hairpin DNA probes. AB - We report a sensitive method for visual detection of mercury ions (II) (Hg2+) in aqueous solution by using gold nanoparticles (Au-NPs) and thymine (T)-rich hairpin DNA probes. The thiolated hairpin DNA probe was immobilized on the Au-NP surface through a self-assembling method. Another thymine-rich, digoxin-labeled DNA probe was introduced to form DNA duplexes on the Au-NP surface with thymine Hg2+-thymine (T-Hg2+-T) coordination in the presence of Hg2+. The Au-NPs associated with the formed duplexes were captured on the test zone of a lateral flow strip biocomponent (LFSB) by immunoreaction events between the digoxin on the duplexes and anti-digoxin antibodies on the LFSB. The accumulation of Au-NPs produced a characteristic red band on the test zone, enabling visual detection of Hg2+ without instrumentation. A detection limit of 0.1 nM was obtained under optimal experimental conditions. This method provides a simple, rapid, sensitive approach for the detection of Hg2+ and shows great promise for point-of-care and in-field detection of environmentally toxic mercury. PMID- 21628096 TI - Au nanospheres and nanorods for enzyme-free electrochemical biosensor applications. AB - Au nanocrystals with different morphologies were prepared and used for enzyme free electrochemical biosensor applications. To investigate the electrocatalytic properties of Au nanocrystals as a function on their morphologies, Au nanocrystals, Au nanospheres (NSs) on silica, Au NSs, and Au nanorods (NRs) with aspect ratios of 1:3 and 1:5, were coated on the screen printed electrodes and further measure the amperometric responses to hydrogen peroxide via three electrode system. The electrodes modified with Au nanocrystals showed biosensing properties without any enzyme being attached or immobilized at their surface. The hydrogen peroxide detection limits of the biosensors with Au NSs, Au NRs (1:3), and Au NRs (1:5) were 6.48, 8.65, and 9.38 MUM (S/N = 3), respectively. The biosensors with Au NSs, Au NRs (1:3), and Au NRs (1:5) showed the sensitivities of 11.13, 54.53, and 58.51 MUA/mM, respectively. These results indicate that morphologies of Au nanocrystals significantly influence the sensitivity of the biosensors. In addition, the enzyme-free biosensors with Au nanocrystals were stable for 2 months. Au nanocrystal-based enzyme-free system, which is proposed in this study, can be used as a platform for various electrochemical biosensors. PMID- 21628097 TI - Modulation of visual physiology by behavioral state in monkeys, mice, and flies. AB - When a monkey attends to a visual stimulus, neurons in visual cortex respond differently to that stimulus than when the monkey attends elsewhere. In the 25 years since the initial discovery, the study of attention in primates has been central to understanding flexible visual processing. Recent experiments demonstrate that visual neurons in mice and fruit flies are modulated by locomotor behaviors, like running and flying, in a manner that resembles attention-based modulations in primates. The similar findings across species argue for a more generalized view of state-dependent sensory processing and for a renewed dialogue among vertebrate and invertebrate research communities. PMID- 21628098 TI - Motoring ahead with rodents. AB - How neural circuits underlie the acquisition and control of learned motor behaviors has traditionally been explored in monkeys and, more recently, songbirds. The development of genetic tools for functional circuit analysis in rodents, the availability of transgenic animals with well characterized phenotypes, and the relative ease with which rats and mice can be trained to perform various motor tasks, make rodents attractive models for exploring the neural circuit mechanisms underlying the acquisition and production of learned motor skills. Here we discuss the advantages and drawbacks of this approach, review recent trends and results, and outline possible strategies for wider adoption of rodents as a model system for complex motor learning. PMID- 21628099 TI - Comparison of 3D spinal motions during stair-climbing between individuals with and without low back pain. AB - In spite of the importance of stair-climbing (SC) as an activity of daily living, 3D spinal motion during SC has not been investigated in association with low back pain (LBP). The purpose of this research is to investigate the differences of the spinal motions during SC between an LBP group and a healthy control group, in order to provide insight into the LBP effect on the spinal motions. During two types of SC tests (single and double step SCs), we measured 3D angular motions (flexion/extension, lateral bending, and twist) of the pelvis, lumbar spine and thoracic spine using an inertial sensing-based, portable spinal motion measurement system. For the nine motion variables (i.e. three anatomical planes * three segments), range of motions (ROM) and movement patterns were compared to determine the differences between the two groups. It was found that the only variable having the p-value of a t-test lower than 0.05 was the flexion/extension of the lumbar spine in both SCs (i.e. the LBP group's ROM 0.215 ng/mL) and 2.80 (95% CI, 1.11 7.10) for the moderate-cotinine group (0.05-0.215 ng/mL), compared with the low cotinine group (< 0.05 ng/mL), adjusting for age, sex, race, educational level, obesity, previous smoking history, job title, and medical history. Plasma cotinine levels were not associated with IL-6 or sICAM-1. CONCLUSIONS: SHS exposure, as assessed by plasma cotinine, was positively associated with hs-CRP in this group of blue-collar workers. The strength of the association with hs-CRP depended on the cut points selected for analysis. PMID- 21628110 TI - New rules proposed for power plant toxics. PMID- 21628111 TI - Public health impact of coal and electricity consumption: risk-benefit balance varies by country. PMID- 21628113 TI - Questions persist: environmental factors in autoimmune disease. PMID- 21628115 TI - Olympic win: lower estimated cancer risk with air pollution controls during the 2008 Beijing Games. PMID- 21628116 TI - Chemical count: quantifying exposures in pregnant women. PMID- 21628117 TI - Pond algae sequester strontium-90. PMID- 21628118 TI - Rotenone and paraquat linked to Parkinson's disease: human exposure study supports years of animal studies. PMID- 21628119 TI - Pollution portrait: the fourth National-Scale Air Toxics Assessment. PMID- 21628120 TI - EHP Classic Paper of the Year, 2011. PMID- 21628121 TI - Nanomaterials and the precautionary principle. PMID- 21628122 TI - Manganese in drinking water and intellectual impairment in school-age children. PMID- 21628123 TI - Coal fire emissions curb children's growth. PMID- 21628124 TI - Investigating human keratinocyte stem cell identity. AB - Studies of the regulatory networks controlling intrinsic properties and fate of adult stem cells are in a large part performed in animal models. Epidermis is one of the most accessible human tissues for researchers, which is a critical parameter for conducting programs dedicated to this knowledge in human stem cell systems. Keratinocyte stem cells constitute a particularly valuable model, because of this practical aspect, but more importantly because their existence is for decades validated by the clinical demonstration of their impressive capacity for epidermis regeneration. For the fundamentalist, human keratinocyte stem cells represent a unique system to dissect the genetic and epigenetic controls of "stemness" and self-renewal. For this purpose, a highly limiting point is our current inability of obtaining a cellular material corresponding to keratinocyte stem cells with homogeneous phenotypic and functional characteristics. The search for tools suitable for the prospective selection of keratinocyte stem cells will benefit from studies conducted at the broad level of the global stem cell field, as well as from more specifically targeted approaches. Advances in that direction are tightly linked to the development of functional assays allowing reliable assessment and modeling of the different stem cell-associated functional characteristics. PMID- 21628125 TI - Cellular adhesion on collagen: a simple method to select human basal keratinocytes which preserves their high growth capacity. AB - The regenerative capacity of human interfollicular epidermis is closely linked to the potential of immature keratinocytes present within its basal layer. The availability of selection methods and culture systems allowing precise assessment of basal keratinocyte characteristics is critical for increasing our knowledge of this cellular compartment. This report presents a multi-parametric comparative study of basal keratinocytes selected according to two different principles: 1) high adhesion capacity on a type-I collagen-coated substrate [Adh+++], 2) high cell-surface expression of alpha6-integrin [Itg-alpha6 (high)]. Importantly, analysis performed at the single-cell level revealed similar primary clone forming efficiency values of 45.5% +/- 6.7% [Itg-alpha6(high)] and 43.7% +/- 7.4% [Adh+++], which were markedly higher than those previously reported. In addition, both methods selected keratinocytes exhibiting an extensive long-term growth potential exceeding 100 cell doublings and the capacity for generating a pluristratified epidermis. Our study also included a global transcriptome comparison. Genome-wide profiling indicated a strong similarity between [Adh+++] and [Itg-alpha6(high)] keratinocytes, and revealed a common basal-associated transcriptional signature. In summary, cross-analysis of [Adh+++] and [Itg alpha6(high)] keratinocyte characteristics showed that these criteria identified highly equivalent cellular populations, both characterized by unexpectedly high growth capacities. These results may have broad impacts in the tissue engineering and cell therapy fields. PMID- 21628126 TI - Epithelial transition zones: merging microenvironments, niches, and cellular transformation. AB - Transition zones (TZs) are regions in the body where two different types of epithelial tissue meet resulting in the appearance of a distinct abrupt transition. These TZs are found in numerous locations within the body, including the cornea-conjunctiva junction, esophagogastric junction, gastro-duodenal junction, endo-ectocervix junction, ileocecal junction, and anorectal junction. Several of these TZs are often associated with the development of cancer, in some cases due to viral transformation by the human papilloma virus (HPV). The underlying molecular and cellular basis for this tumor susceptibiblity is unknown. The distinct epithelial morphology and location results in unique properties being conferred upon this epithelial tissue, as different signaling cues and cell surface markers are apparent. Importantly, the natural state of TZs closely resembles that of a pre-lesional epithelium, as several proteins that are induced during wounding are expressed specifically within this region, which may contribute to transformation. This region may also act as a stem cell niche, and as such, represents a key location for cellular transformation by accumulated genetic mutations or viral transformation resulting in tumor formation. PMID- 21628127 TI - Endocannabinoid signaling and epidermal differentiation. AB - Endocannabinoids represent a class of endogenous lipid mediators, that are involved in various biological processes, both centrally and peripherally. The prototype member of this group of compounds, anandamide, regulates cell growth, differentiation and death; this holds true also in the skin, that is the largest organ of the body constantly exposed to physical, chemical, bacterial and fungal challenges. The epidermis is a keratinized multistratified epithelium that functions as a barrier to protect the organism from dehydration, mechanical trauma, and microbial insults, and epidermal differentiation represents one of the best characterized mechanisms of cell specialization. In this review, we shall summarize current knowledge about the main members of the so-called "endocannabinoid system (ECS)", in order to put in a better perspective the manifold roles that they play in skin pathophysiology. In particular, we shall discuss some aspects of the molecular regulation by endocannabinoids of proliferation and terminal differentiation ("cornification") of mammalian epidermis, showing that ECS is finely regulated by, and can interfere with, the differentiation program. In addition, we shall review evidence demonstrating that disruption of this fine regulation might cause different skin diseases, such as acne, seborrhoea, allergic dermatitis, itch, psoriasis and hair follicle regression (catagen), making of ECS an attractive target for therapeutic intervention. PMID- 21628128 TI - Corneodesmosomes and corneodesmosin: from the stratum corneum cohesion to the pathophysiology of genodermatoses. AB - Corneodesmosin (CDSN) was identified 20 years ago by raising monoclonal antibodies against human plantar stratum corneum. The protein is specific to corneodesmosomes, cell-junction structures that, in humans, are found in the epidermis, the hard palate epithelium, and the inner root sheath of the hair follicles. Synthesized by the granular keratinocytes and secreted via the lamellar bodies, CDSN is incorporated into the desmoglea of the desmosomes, shortly before their transformation into corneodesmosomes during cornification. CDSN displays adhesive properties, mostly attributable to its N-terminal glycine rich domain, and is sequentially proteolyzed as corneocytes migrate towards the skin surface prior to desquamation. The recent inactivation of Cdsn in mice induced a lethal epidermal barrier disruption and hair follicle degeneration, related to corneodesmosome dysfunction. That confirmed the essential role of the protein in maintaining integrity of the epidermis and the hair follicle. The CDSN gene is located in PSORS1, the major psoriasis susceptibility locus on the chromosome 6, but to date its involvement in the disease pathophysiology is not clear. By contrast, two different monogenic diseases associated with nonsense mutations in CDSN, were recently identified. First, hypotrichosis simplex of the scalp in which mutated CDSN accumulates in the dermis and forms amyloid deposits; then, peeling skin disease in which the genetic defect induces dyscohesion of the stratum corneum, responsible for abnormal desquamation and increased skin penetration of allergens. PMID- 21628129 TI - Neuronal sensitivity of the skin. AB - The skin is equipped with nerve fibers subserving the senses for touch, temperature, pain and itch. Thickly myelinated Abeta-fibers are linked to low threshold mechano-receptors responsible to detect vibration and slight indentation of the skin. Among the thinly myelinated Adelta-fibers one class is crucial for the cold detection, but there also nociceptive Adelta-fibers for the detection fast rising noxious heat and punctate mechanical stimuli. Unmyelinated C-fibers consist of various classes mediating nociceptive (pain and itch) and non nociceptive (warmth, pleasant touch) sensations. The unmyelinated C-fibers have close contact to the keratinocytes and the interaction between C-fibers and local skin cells is of particular interest, as efferent neuronal function has both local trophic and immunomodulatory function. In turn, excitability of C-fibers is governed in part by neurotrophins from neighbouring skin cells such as keratinocytes. This mechanism is held responsible to induce and maintain chronic itch and pain conditions. Beyond this interaction recently a direct involvement of keratinocytes in the transduction process has been discussed especially for temperature and noxious stimuli, as keratinocytes express a variety of sensory transduction molecules. Thus, the interplay between neurons and non-neuronal cells is operational not only in basic physiology, but also in chronic itch and pain patients. PMID- 21628130 TI - Psychological stress regulates antimicrobial peptide expression by both glucocorticoid and beta-adrenergic mechanisms. AB - Psychological stress (PS) exerts well-known negative consequences for permeability barrier function in humans and mice, and deterioration of barrier function appears to be attributable largely to excess production of endogenous glucocorticoids (GC). More recently, PS has been shown to compromise antimicrobial defense, also by GC-dependent mechanisms. We assessed here changes in a third antimicrobial peptide (AMP); i.e., the neuropeptide, catestatin (Cst), which also is expressed in the outer epidermis, and previously shown to be regulated by changes in permeability barrier status. In these studies, PS again provoked a decline in both mouse cathelicidin (CAMP) and mouse beta-defensin 3 (mBD3) expression, in a GC-dependent fashion. In contrast, Cst immunostaining instead increased after short-term PS, but then began to decline with more sustained PS. In cultured keratinocytes, we showed further that GC downregulate Cst expression, but beta-adrenergic blockade increased immunostaining for Cst in the face of long-term PS. Furthermore, beta-adrenergic blockade also upregulated CAMP and mBD3 expression. Together, these results suggest that both endogenous GC and beta-adrenergic signaling regulate AMP expression. PMID- 21628131 TI - Development and in vitro assay of oxidative stress modifying formulations for wound healing promotion. AB - Often presented as metabolism byproducts, reactive oxygen species are linked to detrimental effects such as chronic wound, mutagenesis, cancer and skin ageing. However, recent in vitro and in vivo observations suggest that ROS, and mainly hydrogen peroxide, interfere with cell signaling acting like second messenger and inducing adaptive responses. This is particularly observed in skin wound healing where cells are exposed to H2O2 following injury. In this study, we developed and characterized an innovative formulation producing H2O2 at low concentrations, in order to mimic physiological inflammation phase. Then, this pro-oxidative formulation (CAM-GOx) was assayed in vitro on keratinocytes cell culture, compared to the blank formulation (CAM) and the anti-oxidative formulation (CAM CAT) to assess whether oxidative stress was implied or not in cellular responses. PMID- 21628132 TI - Chemometric approach for investigating the skin surface lipids (SSLs) composition: influence of geographical localization. AB - Skin surface lipids (SSLs), arising from both sebaceous glands and skin removal, form a complex lipid mixture composed of free fatty acids and neutral lipids. They are present in the hydrolipidic film and have a close relationship with the stratum corneum lipids. Thus, SSLs participate in the barrier function. One can expect a physiological adaptation of the SSLs composition according to geographical localization or skin pigmentation. In this study, SSLs obtained from three groups of volunteers (light and dark skin, living in France and dark skin, living in Ivory Coast) have been investigated. High-temperature gas chromatography/mass spectrometry has been used to study SSLs composition. The variability in the SSLs chromatographic profiles has been investigated, using chemometric methods which allowed us to highlight the sapienic acid (C16: 1Delta6) as discriminant according to geographical localization. This result is of a great interest regarding sapienic acid properties. However, no significant variation has been detected following skin pigmentation. In parallel, SSLs Raman spectra were collected in order to determine the organization and the conformational order of lipids. The spectral data treatment revealed discriminant variation in the upsilon C-C stretching region revealing changes in the conformational order of the SSLs following geographical localization. PMID- 21628133 TI - Skin physiology: crossroads for a multidisciplinary science. PMID- 21628134 TI - Corpus callosotomy with gamma knife radiosurgery for a case of intractable generalised epilepsy. AB - Gamma knife radiosurgery is a minimally invasive procedure which can be used for patients with intractable epilepsies as an alternative for surgical corpus callosotomy. We report a 13-year-old boy with intractable epilepsy who underwent radiosurgical callosotomy. The patient demonstrated significant clinical improvement after gamma knife radiosurgery and was free of seizures 10 months after the procedure. However, He developed four short focal seizures with clonic movements during the 20 months post radiosurgery. Corpus callosotomy decreased epileptiform discharges in both hemispheres, indicating a role for the callosal neurons to facilitate an asymmetric epileptogenic susceptible state within the two hemispheres such that bisynchronous and bisymmetrical epileptiform discharges develop. Our result demonstrates that this novel therapeutic approach is a safe and effective option for the treatment of intractable generalised epilepsies. PMID- 21628135 TI - Pro- and anti-inflammatory responses are regulated simultaneously from the first moments of septic shock. AB - The relationships between cytokine responses in septic shock are currently poorly understood. Some studies have pointed to a biphasic model, with an initial proinflammatory phase, followed by a reactive, anti-inflammatory response to explain the pathogenesis of the most severe form of sepsis. However, evidence for the coexistence of both responses has been found. In this study, the plasma levels of 17 cytokines and chemokines, in 20 patients with septic shock, 11 patients with systemic inflammatory response syndrome (SIRS), during the first 24 hours following diagnosis, and 10 healthy controls, were analyzed and compared. Patients with septic shock showed increased levels of IL-6, IL-8, MCP-1, MIP 1beta, IFN-gamma, GM-CSF and IL-10 compared to healthy controls. Patients with SIRS showed higher levels of IL-6, IL-8, MCP-1, MIP-1beta, G-CSF and IL-10 than controls. Patients with septic shock showed higher levels of IL-8, GM-CSF, MIP 1beta than those with SIRS. The Spearman test demonstrated a positive association between the pro-inflammatory mediators IL-6, IL-8, MCP-1, MIP-1beta, IFN-gamma, GM-CSF and the immunomodulatory cytokine IL-10 in septic shock. Consequently, correlation studies supported the notion that secretion of pro- and anti inflammatory mediators in septic shock occurs as a simultaneous immune response program initiated early in the course of the disease, revealing that both types of cytokine play a role from the very beginning of this life-threatening condition. PMID- 21628136 TI - Benign childhood seizure susceptibility syndrome: three case reports. AB - In this study, we describe three patients who each had two different forms of idiopathic focal epilepsiy. Two of these patients had electroclinical features compatible with Panayiotopoulos syndrome and benign childhood epilepsy with centro-temporal spikes (BCECTS), one of whom developed a particular electroclinical picture of atypical benign focal epilepsy and the other an atypical evolution characterized by verbal auditory agnosia and aphasia. The third patient had clinical and electroencephalographic features of BCECTS and of idiopathic childhood occipital epilepsy (Gastaut type) which evolved into electroclinical features of continuous spikes and waves during slow sleep (CSWS). All three patients presented with two focal idiopathic epilepsies with a particular evolution associated with CSWS, supporting the concept of benign childhood seizure susceptibility syndrome as described by Panayiotopoulos (1993). PMID- 21628137 TI - Reducing patient harm with the use of opioid oral solutions. AB - Medication errors involving oral opioids have led to serious adverse events, including failure to control pain, over-sedation, respiratory depression, seizures, and death. This article will highlight medication errors that may arise during the use of opioid oral solutions, particularly concentrated formulations, and provide risk-reduction recommendations to address the potential for error and patient harm. PMID- 21628138 TI - Care of the elderly patient with prostate cancer: what's new and what's the same. AB - As the number of men in the United States 65 years of age or older increases- from 12% in 2000 to 20% in 2030--the burden of prostate cancer also is increasing. Standard therapy is appropriate for healthy men with a life expectancy of more than 10 years; however, clinicians may fear either overtreating frail patients or undertreating patients who are physically fit. Evidence suggests the best approach is to conduct a comprehensive geriatric assessment using available measures and published guidelines to individualize treatment plans. Recent Food and Drug Administration approval of cabazitaxel, sipuleucel-T, and denosumab has expanded available treatment options. Managing symptoms related to disease complications and side effects of treatment is increasingly more complicated with the addition of these new regimens and extended life expectancy. Pharmacists' participation in patient assessment and supportive care are necessary components of comprehensive care for cancer patients. PMID- 21628139 TI - Building a pharmacist-managed pharmacotherapy medication therapy management practice. AB - OBJECTIVE: To describe the development and growth of a pharmacotherapy medication therapy management (MTM) practice. SETTING: Multispecialty medical clinic. PRACTICE DESCRIPTION: MTM practice based in a multispecialty medical clinic with a community pharmacy. The practice serves primarily internal medicine, family medicine, bariatric surgery, and solid-organ transplant surgery patients that are referred by clinic providers. The practice serves 30 to 40 patients per month and employs approximately one pharmacist spending 50% to 75% of his/ her time performing MTM. PRACTICE INNOVATION: This MTM practice serves both primary care and specialty care patients via different practice models in a physician office setting. Unlike many MTM practices, this practice is financially self-sustaining. MAIN OUTCOME MEASUREMENTS: Sustainability, continued growth, financial viability. RESULTS: After early slow growth, the pharmacotherapy MTM practice at Mayo Clinic Florida has grown into multiple practice models. CONCLUSION: Institutions can use this practice model as a guide to assist them in the creation of future institutional pharmacotherapy MTM models. PMID- 21628140 TI - Medication-induced sleep disturbances. AB - OBJECTIVE: To review the literature for medication-induced sleep disturbances. DATA SOURCE: MEDLINE/PUBMED searches (January 2000-May 2010) were conducted to identify pertinent English-language studies. STUDY SELECTION AND DATA EXTRACTION: References were reviewed to identify literature that provided evidence of sleep disturbances caused by prescription medications. DATA SYNTHESIS: Review of studies was performed to determine the effect a given medication had on subjects' sleep architecture or those that induced common sleep disorders. Tables were constructed for specific sleep disturbances to guide clinician recognition of such effects. CONCLUSION: A wide array of prescription medications used in the management of medical and psychiatric disorders can affect an individual's sleep patterns. Different modalities of sleep disturbances may be affected by a multitude of prescription medications. This review serves as an updated description of medications altering sleep architecture or inducing insomnia, periodic limb movement, sedation, sleep attacks, nightmares, and REM parasomnias. To improve patient outcomes, a consultant pharmacist should be familiar with medication-induced sleep disturbances. PMID- 21628141 TI - Insulin pens vs. vials and syringes: differences in clinical and economic outcomes. AB - Insulin therapy is crucial to the treatment of many patients with diabetes mellitus. Therefore, it is important for pharmacists to consider the clinical and economic implications of the decision between insulin pens and insulin vials and syringes. Clinically, insulin pens show an advantage through improved adherence and reduced hypoglycemic events. Furthermore, overall health care costs were either unchanged or improved in insulin pen users as compared with those using insulin vials and syringes, although little economic advantage was observed when switching from insulin vials to insulin pens. Patients tend to prefer insulin pen use based on patient satisfaction and ease of use. Through an understanding of the advantages and disadvantages of insulin pens and vials and syringes, pharmacists can help to advocate for the most appropriate insulin-delivery method to maximize clinical outcomes and to reduce overall health care spending. PMID- 21628142 TI - Bedbugs, other infestations, and outbreaks: no magic bullet. PMID- 21628143 TI - Anatomy of axis contraction in seedlings from a fire prone habitat. AB - The hypocotyls or roots of many seed plants contract during seedling growth. Anatomical evidence is here reported for the first time that G-fibers (gelatinous or tension wood fibers) may cause contraction of roots and hypocotyls in dicotyledonous seedlings long after germination. To document repositioning of seedling buds, selected perennials (20 dicotyledons and one cycad) native to the fire-prone pine rocklands of subtropical South Florida were germinated and measured for 4-5 mo. The height of cotyledonary nodes above the soil decreased because of axis contraction or bending in eight species. Anatomy suggested that two mechanisms operate: (1) previously well-documented collapse of parenchyma cells in two species (Convolvulaceae and Zamiaceae) and (2) newly documented production of G-fibers in six species (all Fabaceae). Contraction or bending of the hypocotyl and/or taproot moved the cotyledonary and later buds of the seedling closer to the soil surface or buried them. Bud repositioning by these mechanisms may protect the lateral buds from injury by fire or other environmental stresses and allow resprouting. PMID- 21628144 TI - Towards unlocking the deep nodes of Leguminosae: Floral development and morphology of the enigmatic Duparquetia orchidacea (Leguminosae, Caesalpinioideae). AB - Duparquetia orchidacea (Caesalpinioideae-Cassieae-Duparquetiinae) is a monotypic liana from tropical West Africa. Its highly unusual, zygomorphic flowers, the unique pollen morphology, and the lack of vestured pits in the wood correspond with previous phylogenetic studies that resolved the position of the species to an isolated position among the early-branching Leguminosae. Here we present a detailed analysis of floral morphology and development to clarify open questions of its floral organization. We provide new data that can be useful in clarifying phylogenetic relationships among early branching Leguminosae and improve our understanding of floral evolution in this large and important plant family. For comparison, we also present developmental data for other Fabales. Our analysis reveals some unusual and in parts unique developmental patterns, such as strict acropetal organ formation, loss and suppression of floral organs, and early petal enlargement. We interpret alternating left-right symmetries in floral development as clues to a spiral organ formation in ancestral taxa. Early asymmetry of the young carpel helps to interpret enantiostyly of other Leguminosae as an example of imprinted shape. Finally, we show that cochlear-descending petal aestivation in Duparquetia and in Papilionoideae is based on different ontogenetic patterns and therefore is most probably nonhomologous. PMID- 21628145 TI - Do bryophyte shoot systems function like vascular plant leaves or canopies? Functional trait relationships in Sphagnum mosses (Sphagnaceae). AB - Vascular plant leaf traits that influence photosynthetic function form the basis of mechanistic models of carbon exchange. Given their unique tissue organization, bryophytes may not express similar patterns. We investigated relationships among tissue, shoot, and canopy traits, and their associations with photosynthetic characteristics in 10 Sphagnum species. Trait relationships were organized around a primary dimension accounting for 43% of variation in 12 traits. There was no significant relationship between nitrogen content of shoot systems and maximum photosynthesis expressed on mass (A(mass)) or area (A(area)) bases due to nitrogen sequestration and storage within the canopy interior. This pattern differs from the distribution of nitrogen in vascular plant canopies. Thus, nitrogen and its relationship to carbon uptake in Sphagnum shoots does not conform to patterns of either vascular plant leaves or canopies. Species that concentrate biomass and nitrogen in the capitulum have enhanced rates of A(mass) and A(area). Consequently, A(area) was positively associated with N(area) of the capitulum only. Overall, water content and carotenoid concentration were the strongest predictors of both A(mass) and A(area) and these were expressed as inverse relationships. The relationships of plant traits in Sphagnum defines a principal trade-off between species that tolerate environmental stress and those that maximize carbon assimilation. PMID- 21628146 TI - Plant-pollinator interactions in tropical monsoon forests in Southeast Asia. AB - Forests with different flora and vegetation types harbor different assemblages of flower visitors, and plant-pollinator interactions vary among forests. In monsoon dominated East and Southeast Asia, there is a characteristic gradient in climate along latitude, creating a broad spectrum of forest types with potentially diverse pollinator communities. To detect a geographical pattern of plant pollinator interactions, we investigated flowering phenology and pollinator assemblages in the least-studied forest type, i.e., tropical monsoon forest, in the Vientiane plain in Laos. Throughout the 5-year study, we observed 171 plant species blooming and detected flower visitors on 145 species. Flowering occurred throughout the year, although the number of flowering plant species peaked at the end of dry season. The dominant canopy trees, including Dipterocarpaceae, bloomed annually, in contrast to the supra-annual general flowering that occurs in Southeast Asian tropical rain forests. Among the 134 native plant species, 68 were pollinated by hymenopterans and others by lepidopterans, beetles, flies, or diverse insects. Among the observed bees, Xylocopa, megachilids, and honeybees mainly contributed to the pollination of canopy trees, whereas long-tongued Amegilla bees pollinated diverse perennials with long corolla tubes. This is the first community-level study of plant-pollinator interactions in an Asian tropical monsoon forest ecosystem. PMID- 21628147 TI - Leaf phenology in relation to canopy closure in southern Appalachian trees. AB - Leaf phenology varies markedly across tree species of temperate deciduous forests. Early leafing in spring may increase light capture and carbon gain prior to canopy closure, allowing saplings to survive in understory sites deeply shaded in midsummer. We quantified sapling leaf phenology for 18 tree species and seasonal variation in understory light availability at three sites along a ridge slope-cove landform gradient in the Great Smoky Mountains National Park. Early leafing species (e.g., Aesculus flava, Carpinus caroliniana) broke bud an average of 24 d before late leafers (e.g., Magnolia fraseri, Nyssa sylvatica). Canopy closure occurred 14-18 d earlier and summer understory light was on average 63 74% lower on intermediate and mesic sites than on the xeric site. Early leafing species intercepted 45-80% of their growing season photon flux before canopy closure vs. 8-15% for late leafers. However, earlier leafing increased exposure to freezing temperatures by 5.5% per week near the mean time of bud break. Early leafing is strongly correlated with midsummer shade, risk of freezing temperatures, and distribution on mesic sites across a "main spectrum" of 15 deciduous species. Differences in leaf phenology and resultant impacts on spring carbon gain may help determine tree shade tolerance and distribution in southern Appalachian forests. PMID- 21628148 TI - Opportunistic nectar-feeding birds are effective pollinators of bird-flowers from Canary Islands: experimental evidence from Isoplexis canariensis (Scrophulariaceae). AB - Insular floras, characterized by simple pollination networks, sometimes include novel mutualistic agents such as nonspecialist nectarivores. In this study we confirmed the effective pollination of Isoplexis canariensis by opportunistic nectar-feeding birds in Tenerife, Canary Islands, Spain. This plant is among the ornithophilous species of the Canarian flora that lack past and present specialist nectarivorous birds. Experimental hand pollinations revealed self compatibility, but cross-pollinated flowers produced a greater percentage of viable seeds than self-pollinated ones. Flowers were visited by five species of birds (Phylloscopus canariensis, Parus caeruleus, Sylvia melanocephala, Serinus canarius, and Fringilla coelebs) and by the endemic lizard (Gallotia galloti, Lacertidae). Insect pollination was absent, and the few insect visitors acted as nectar thieves or secondary nectar robbers. Birds represented 93.1% of total visits, with the Canarian Chiffchaff, Ph. canariensis, being the most frequent visitor. Flowers visited by birds set more, larger, and heavier fruit than flowers from which birds were excluded. Bird visitation also enhanced seed viability. These results demonstrate the active role of these opportunistic birds as effective pollinators of this Canarian bird-flower species. Further, the results reveal the need to consider the effect of these birds on the evolution of ornithophilous floral traits in absence of specialist nectarivores. PMID- 21628149 TI - Effects of Alliaria petiolata (garlic mustard; Brassicaceae) on mycorrhizal colonization and community structure in three herbaceous plants in a mixed deciduous forest. AB - Herbaceous plant species are important components of forest ecosystems, and their persistence in forests may be affected by invasive plant species that reduce mycorrhizal colonization of plant roots. I examined the effect of the invasive plant Alliaria petiolata on arbuscular mycorrhizal fungi (AMF) colonizing the roots of three forest plant species. AMF root colonization and community structure was examined from plants that were growing either in the absence or presence of Alliaria under natural forest conditions. AMF root colonization varied among the plant species but was not significantly affected by Alliaria. With molecular methods, ~12 different taxa of AMF could be distinguished among the root samples, and these taxa belonged to the genera Acaulospora and Glomus, with Glomus dominating AMF communities. There were significant differences between the community of AMF colonizing roots of Maianthemum racemosum and Trillium grandiflorum, but only AMF communities of Maianthemum roots were significantly affected by Alliaria. Indicator species analysis found that an Acaulospora species type was a significant indicator of Maianthemum plants grown in the absence of Alliaria. These results suggest invasive plants like Alliaria may selectively suppress AMF fungi, and this suppression can affect AMF communities colonizing the roots of some native plant species. PMID- 21628150 TI - Phylogenetic comparative analysis of microsporogenesis in angiosperms with a focus on monocots. AB - This paper presents the first broad overview of three main features of microsporogenesis (male meiosis) in angiosperms: cytokinesis (cell division), intersporal wall formation, and tetrad form. A phylogenetic comparative approach was used to test for correlated evolution among these characters and to make hypotheses about evolutionary trends in microsporogenesis. The link between features of microsporogenesis and pollen aperture type was examined. We show that the pathway associated with successive cytokinesis (cytoplasm is partitioned after each meiotic division) is restricted to wall formation mediated by centrifugally developing cell plates, and tetragonal (or decussate, T-shaped, linear) tetrads. Conversely, much more flexibility is observed when cytokinesis is simultaneous (two meiotic divisions completed before cytoplasmic partitioning). We suggest that the ancestral type of microsporogenesis for angiosperms, and perhaps for all seed plants, associated simultaneous cytokinesis with centripetal wall formation, resulting in a large diversity in tetrad forms, ranging from regular tetrahedral to tetragonal tetrads, including rhomboidal tetrads. From this ancestral pathway, switches toward successive cytokinesis occurred among basal angiosperms and monocots, generally associated with a switch toward centrifugal intersporal wall formation, whereas eudicots evolved toward an almost exclusive production of regular tetrahedral tetrads. No straightforward link is found between the type of microsporogenesis and pollen aperture type. PMID- 21628151 TI - Bidirectional history of hybridization in California wild radish, Raphanus sativus (Brassicaceae), as revealed by chloroplast DNA. AB - The evolutionary processes that take place in invasive plant populations are not well documented or understood. Interspecific hybridization between cultivated radish (Raphanus sativus) and R. raphanistrum is known to be responsible for the origin of the invasive California wild radish, but little is known about the nature of the hybridization events that produced the hybrid-derived lineage. We analyzed the trnL-rpl32 intergenic region of chloroplast DNA (cpDNA) obtained from 37 cultivated radish individuals from four different cultivars, 53 R. raphanistrum individuals from six European populations and 104 California wild radish individuals from 11 populations covering its entire range throughout the state. We found that cultivated radish and R. raphanistrum shared no cpDNA haplotypes but that they both shared haplotypes with California wild radish, evidence for bidirectional hybridization between the progenitor species in the creation of the California lineage. We also found evidence that multiple cultivars and multiple European source populations contributed to the diversity of cpDNA haplotypes within California. Studies like this will continue to be important for our understanding of the origin of invasive populations and the mechanisms by which they succeed. PMID- 21628152 TI - Development and ecological implications of dormant buds in the high Paleolaltitude Triassic sphenophyte Spaciinodum (Equisetaceae). AB - Spaciinodum collinsonii, a Triassic sphenophyte from the central Transantarctic Mountains, Antarctica, is reinterpreted based on new material in order to clarify discrepancies from previous work and to detail the development and ecology of the Spaciinodum plant. Vegetative stems have alternating nodes and internodes, nodes distinguished by a solid diaphragm of tissue, internodes by the presence of vallecular (cortical) and carinal canals, and a hollow pith. Whorls of branches arise immediately above the nodes, alternating with the leaves of the subjacent nodes. Branches develop in the cortex and are anatomically similar to the stems. While Spaciinodum is similar to extant Equisetum, it is distinctive in that its large vallecular canals form a complete ring within the cortex and are separated only by thin fimbrils of tissue. The majority of specimens of Spaciinodum are now believed to be dormant buds with condensed nodes and internodes, with progressively longer internodal regions more basally. More apical portions of buds have cellular internodes because the areas where the canals will form have not yet ruptured from elongation. The abundance of buds and the absence of elongated stems in the permineralized peat deposit suggest that Spaciinodum underwent dormancy during the dark Antarctic winters. PMID- 21628153 TI - At the interface of phylogenetics and population genetics, the phylogeography of Dirca occidentalis (Thymelaeaceae). AB - Dirca occidentalis is a rare shrub indigenous to only six counties near the San Francisco Bay in California, United States. We used intersimple sequence repeat (ISSR) markers and automated genotyping to probe 29 colonies of D. occidentalis from four geographically disjunct populations (East Bay, North Bay, Salmon Creek, and Peninsula) and used methods of phylogenetics and population genetics to model variation across the species. Results show that the four disjunct populations are genetically isolated and have undergone divergence. Phylogenetic analyses indicate that the East Bay population was the first to diverge, followed by the North Bay, then the Salmon Creek and Peninsula populations. This order of divergence suggests an intriguing natural history for D. occidentalis that is explained by the dynamic geological and climatic history of the Bay Area. Spatial genetic structure detected for the species suggests an interaction of four factors: limited seed dispersal, clonal regeneration, distances traveled by pollinators, and genetic isolation of the four populations. Genetic diversity within the North Bay and Salmon Creek populations is low, indicating poor ecological fitness and risk of decline. ISSRs resolved phylogeographic structure within D. occidentalis, results unattainable with ITS methods, and the integration of tools of phylogenetics and population biology led to an enhanced understanding of this endemic species. PMID- 21628154 TI - Nei's to Bayes': comparing computational methods and genetic markers to estimate patterns of genetic variation in Tolpis (Asteraceae). AB - Accurate determination of patterns of genetic variation provides a powerful inferential tool for studies of evolution and conservation. For more than 30 years, enzyme electrophoresis was the preferred method for elucidating these patterns. As a result, evolutionary geneticists have acquired considerable understanding of the relationship between patterns of allozyme variation and aspects of evolutionary process. Myriad molecular markers and statistical analyses have since emerged, enabling improved estimates of patterns of genetic diversity. With these advances, there is a need to evaluate results obtained with different markers and analytical methods. We present a comparative study of gene statistic estimates (F(ST), G(ST), F(IS), H(S), and H(T)) calculated from an intersimple sequence repeat (ISSR) and an allozyme data set derived from the same populations using both standard and Bayesian statistical approaches. Significant differences were found between estimates, owing to the effects of marker and analysis type. Most notably, F(ST) estimates for codominant data differ between Bayesian and standard approaches. Levels of statistical significance are greatly affected by methodology and, in some cases, are not associated with similar levels of biological significance. Our results suggest that caution should be used in equating or comparing results obtained using different markers and/or methods of analysis. PMID- 21628155 TI - Embryology of Koeberlinia (Koeberliniaceae): Evidence for core-Brassicalean affinities. AB - Koeberlinia, comprising a single xerophytic species K. spinosa, had previously been placed in various families, mainly Capparaceae. Current molecular evidence now places it in its own family Koeberliniaceae, thought to be related to the Bataceae/Salvadoraceae among the 17 other families of the Brassicales. We investigated 55 embryological characters of the genus, most of which are not understood yet, and thereby assessed its systematic relationships. Koeberlinia has many embryological features in common with the Capparaceae and seven other core-Brassicalean families (i.e., Brassicaceae, Cleomaceae, Emblingiaceae, Gyrostemonaceae, Pentadiplandraceae, Resedaceae, and Tovariaceae), specifically by possessing a campylotropous ovule with a nonmultiplicative (two-cell-layered) outer integument, reniform seeds with a curved embryo, and a fibrous exotegmen in the mature seed coat. However, Koeberlinia is clearly distinguished from them by a tenuinucellate rather than crassinucellate ovule as previously reported, markedly enlarged apical nucellar epidermal cells, and an "exotestal" seed coat. Embryologically, Koeberlinia resembles neither the Bataceae nor the Salvadoraceae, although only limited embryological data are available for these two families. Embryological evidence thus favors its joining the core Brassicales, but additional molecular analyses and embryological studies on the missing data of the Bataceae and Salvadoraceae are needed for final confirmation of its phylogenetic position. PMID- 21628156 TI - Intercourse between cell wall and cytoplasm exemplified by arabinogalactan proteins and cortical microtubules. AB - How does a plant cell sense and respond to the status of its cell wall? Intercourse between cell wall and cytoplasm has long been supposed to involve arabinogalactan proteins, in part because many of them are anchored to the plasma membrane. Disrupting arabinogalactan proteins has recently been shown to disrupt the array of cortical microtubules present just inside the plasma membrane, implying that microtubules and arabinogalactan proteins interact. In this article, we assess possibilities for how this interaction might be mediated. First, we consider microdomains in the plasma membrane (lipid rafts), which have been alleged to link internal and external regions of the plasma membrane; however, the characteristics and even the existence of these domains remains controversial. Next, we point out that disrupting the synthesis of cellulose also can disrupt microtubules and consider whether arabinogalactan proteins are part of a network linking microtubules and nascent microfibrils. Finally, we outline several signaling cascades that could transmit information from arabinogalactan proteins to microtubules through channels of cellular communication. These diverse possibilities highlight the work that remains to be done before we can understand how plant cells communicate across their membranes. PMID- 21628157 TI - Wound-induced vascular occlusions in Vitis vinifera (Vitaceae): Tyloses in summer and gels in winter1. AB - Vascular occlusion in xylem conduits is a common response to environmental stresses, and plant species are recognized as primarily tylose-forming or gel forming. These stresses occur throughout the year, but there is little information on the wound responses throughout the year and in growing and dormant tissues. Wound-induced vascular occlusions were evaluated by type (tylose or gel), temporal progress, and spatial distribution for grape stems pruned in four seasons through an entire year. Tyloses were formed predominantly in summer and gels in winter. Cytohistological analyses indicated that wound-induced gels were pectin-rich. Both gel formation and tylose development were complete within 7 d and 10 mm from the cut regardless of the season of the wounding. Most vessels were affected by wounding, but a higher fraction of vessels developed occlusions in summer and autumn (over 80%) than in winter and spring (about 60%). The study is the first to show a single species is capable of producing primarily either tyloses or gels and that the type of wound-induced occlusion is dependent upon the season in which wounding occurs. Winter conditions limit the wound response to reversible gel formation that may contribute to refilling of embolized vessels in the spring. PMID- 21628158 TI - Root anatomy, morphology, and longevity among root orders in Vaccinium corymbosum (Ericaceae). AB - Understanding root processes at the whole-plant or ecosystem scales requires an accounting of the range of functions within a root system. Studying root traits based on their branching order can be a powerful approach to understanding this complex system. The current study examined the highly branched root system of the ericoid plant, Vaccinium corymbosum L. (highbush blueberry) by classifying its root orders with a modified version of the morphometric approach similar to that used in hydrology for stream classification. Root anatomy provided valuable insight into variation in root function across orders. The more permanent portion of the root system occurred in 4th- and higher-order roots. Roots in these orders had radial growth; the lowest specific root length, N:C ratios, and mycorrhizal colonization; the highest tissue density and vessel number; and the coarsest root diameter. The ephemeral portion of the root system was mainly in the first three root orders. First- and 2nd-order roots were nearly anatomically identical, with similar mycorrhizal colonization and diameter, and also, despite being extremely fine, median lifespans were not very short (115-120 d; estimated with minirhizotrons). Our research underscores the value of examining root traits by root order and its implications to understanding belowground processes. PMID- 21628159 TI - Anatomical and histochemical characterization of extrafloral nectaries of Prockia crucis (Salicaceae). AB - Besides being vital tools in taxonomic evaluation, the anatomy of plant secretory structures and the chemical composition of their secretions may contribute to a more thorough understanding of the roles and functions of these secretory structures. Here we used standard techniques for plant anatomy and histochemistry to examine secretory structures on leaves at different stages of development of Prockia crucis, to evaluate the origin and development of the structures, and to identify the disaccharides and monosaccharides in the exudates. Fructose, glucose, and sucrose constituted up to 49.6% of the entire secretion. The glands were confirmed to be extrafloral nectaries (EFNs); this is the first report of their presence in the genus Prockia. These EFNs are globular, sessile glands, with a central concavity occurring on the basal and marginal regions of the leaf. The epidermis surrounding the concavity is secretory, forming a single-layered palisade that strongly reacts with periodic acid-Schiff's reagent (PAS) and xylidine Ponceau, indicators of total polysaccharides and total proteins, respectively, in the exudate. On the basis of the similarity of these glands to the salicoid teeth in Populus and Salix, we suggest that these three taxa are phylogenetically close. PMID- 21628160 TI - A scaling law for the effects of architecture and allometry on tree vibration modes suggests a biological tuning to modal compartmentalization. AB - Wind is a major ecological factor for plants and a major economical factor for forestry. Mechanical analyses have revealed that the multimodal dynamic behavior of trees is central to wind-tree interactions. Moreover, the trunk and branches influence dynamic modes, both in frequency and location. Because of the complexity of tree architecture, finite element models (FEMs) have been used to analyze such dynamics. However, these models require detailed geometric and architectural data and are tree-specific-two major restraints for their use in most ecological or biological studies. In this work, closed-form scaling laws for modal characteristics were derived from the dimensional analysis of idealized fractal trees that sketched the major architectural and allometrical regularities of real trees. These scaling laws were compared to three-dimensional FEM modal analyses of two completely digitized trees with maximal architectural contrast. Despite their simplifying hypotheses, the models explained most of the spatiotemporal characteristics of modes that involved the trunk and branches, especially for sympodial trees. These scaling laws reduce the tree to (1) a fundamental frequency and (2) one architectural and three biometrical parameters. They also give quantitative insights into the possible biological control of wind excitability of trees through architecture and allometries. PMID- 21628161 TI - The climbing habit in palms: Biomechanics of the cirrus and flagellum. AB - Climbing palms in the Arecoideae (Desmoncus) and Calamoideae (rattan palms) both evolved cirrate leaves armed with hooks and grapnels for climbing. Some species of Calamoideae develop a different climbing organ known as the flagellum, which also bears hooks. The present study indicates that geometry and mechanical properties of the cirrus vary between species. Cirrate leaves are constructed to optimize bending and torsion in relation to the deployment of recurved hooks. Hook development, size, and strength vary along cirri and flagella and are consistent with observations of these attachment organs functioning as a ratchet mechanism: hooks increase in strength toward the base of attachment organs and always fail before the axis in strength tests. Hook size and strength differ between species and are related to body size and ecological preference. Larger species produce larger hooks, but smaller climbing palms of the understory deploy fine sharp hooks that are effective on small diameter supports as well as large branches and trunks. The ephemeral nature of climbing organs in palms provides a challenge to their life-history development, particularly in terms of mechanical constraints and remaining attached to the host vegetation; these differ significantly from many vines and lianas having more perennial modes of attachment. PMID- 21628162 TI - A new lineage of lichenized basidiomycetes inferred from a two-gene phylogeny: The Lepidostromataceae with three species from the tropics. AB - The lichen habit has apparently evolved independently in at least five major clades of mushroom-forming basidiomycetes (Agaricomycetes). Tracing the origin of lichenization in these groups depends on a clearer understanding of the phylogenetic relationships of basidiolichens to other fungi. We describe here a new family of basidiolichens made up of tropical, soil-inhabiting fungi that form lichenized, scale-like squamules and erect, coral-like fruiting structures. These structures are common to two basidiolichen genera, Multiclavula and Lepidostroma. Molecular studies have confirmed the phylogenetic position of Multiclavula species in the Cantharellales, but Lepidostroma species have never been sequenced. We obtained nuclear small and large subunit ribosomal sequences from specimens of L. calocerum collected in Costa Rica and Mexico and also from specimens of two Multiclavula spp. recently described from Rwanda. The phylogenetic placement of these fungi within the Agaricomycetes was investigated using likelihood and Bayesian analyses. Our results indicate that L. calocerum and the Rwandan species form a natural group unrelated to Multiclavula and sister to the Atheliales, members of which are neither lichen-forming nor clavarioid. The independent evolution of morphologically similar forms in so many groups of basidiomycetes is a remarkable example of convergence, indicating similar pathways to lichenization in these fungi. PMID- 21628163 TI - Importance of plant traits and herbivory for invasiveness of Phragmites australis (Poaceae). AB - Biological invasions change native plant communities, but theory predicting whether introductions create naturalized or invasive species is lacking. Focusing on either plant traits or interactions of introduced plants with native biota creates unreliable results, and improvements may require integration of trait- and interaction-based approaches. To assess the importance of plant traits and herbivory on invasiveness, we incorporated herbivore effects in comparisons of growth and phenology of invasive Phragmites australis and its native congener P. australis subsp. americanus. Our results were influenced by venue (field or common garden), with extended life span and optimized leaf-age structure of introduced P. australis indicating greater potential for resource capture. Attack by introduced gallflies affected expression of plant traits, but we found no consistent effect of aphid attack. Origin did not affect leaf emergence or stem height, but preferential gallfly attack stunted native P. australis and delayed senescence. Greater resource capture and lower attack by nonnative herbivores could give introduced P. australis an advantage over the native subspecies. Our results demonstrating the importance of plant traits as well as their modification by interactions with natural enemies questions whether the outcome of plant introductions can be predicted. PMID- 21628164 TI - Fossil Araceae from a Paleocene neotropical rainforest in Colombia. AB - Both the fossil record and molecular data support a long evolutionary history for the Araceae. Although the family is diverse in tropical America today, most araceous fossils, however, have been recorded from middle and high latitudes. Here, we report fossil leaves of Araceae from the middle-late Paleocene of northern Colombia, and review fossil araceous pollen grains from the same interval. Two of the fossil leaf species are placed in the new fossil morphogenus Petrocardium Herrera, Jaramillo, Dilcher, Wing et Gomez-N gen. nov.; these fossils are very similar in leaf morphology to extant Anthurium; however, their relationship to the genus is still unresolved. A third fossil leaf type from Cerrejon is recognized as a species of the extant genus Montrichardia, the first fossil record for this genus. These fossils inhabited a coastal rainforest ~60-58 million years ago with broadly similar habitat preferences to modern Araceae. PMID- 21628165 TI - Bromus tectorum (Poaceae) in midcontinental United States: Population genetic analysis of an ongoing invasion. AB - Biological invasions can be substantially influenced by the genetic sampling associated with a species' introduction. As a result, we assessed the genetic and evolutionary consequences of the entry and spread of the invasive grass Bromus tectorum (cheatgrass) across the United States midcontinent through an analysis of 54 populations, using enzyme electrophoresis. On average, these populations display 1.04 alleles per locus (A), 4.1% percent polymorphic loci per population (%P) and an expected mean heterozygosity (H(exp)) value of 0.009. Heterozygotes, which have been rarely reported for B. tectorum in North America, occur in three populations in the midcontinent and are likely novel multilocus genotypes that arose postimmigration. The midcontinent distribution of multilocus genotypes suggests that plant immigrants came directly from either the native range or the eastern United States, or both. Continued dispersal of preadapted genotypes and the assembly of populations that are genetic admixtures may enhance this invasion by increasing both the genetic diversity within populations and the selection of novel genotypes arising from occasional outcrossing. The potential for postimmigration evolution in most species points to the largely unrecognized need to block the introduction of new, potentially aggressive genotypes of an alien species already in the United States. PMID- 21628166 TI - The inbreeding depression cost of selfing: Importance of flower size and population size in Collinsia parviflora (Veronicaceae). AB - Inbreeding depression should evolve with selfing rate when frequent inbreeding results in exposure of and selection against deleterious alleles. The selfing rate may be modified by plant traits such as flower size, or by population characteristics such as census size that can affect the probability of biparental inbreeding. Here we quantify inbreeding depression (delta) among different population sizes of Collinsia parviflora, a wildflower with interpopulation variation in flower size, by comparing fitness components and multiplicative fitness of experimentally produced selfed and outcrossed offspring. Selfed offspring had reduced multiplicative fitness compared to outcrossed offspring, but inbreeding depression was low in all combinations of population size and flower size (delta <= 0.05) except in large populations of large-flowered plants (delta = 0.45). The decrement to multiplicative fitness with inbreeding was not affected by population size nested within flower size, but differed between small and large-flowered plants: small-flowered populations had lower overall inbreeding depression (delta = 0.04) compared to large-flowered populations (delta = 0.25). The difference in load with flower size suggests that either selection has removed deleterious recessive alleles or these alleles have become fixed in small-flowered, potentially more selfing populations, but that purging has not occurred to the same extent in presumably outcrossing large-flowered populations. PMID- 21628167 TI - Cytotype variation and allopolyploidy in North American species of the Sphagnum subsecundum complex (Sphagnaceae). AB - Allopolyploid speciation is likely the predominant mode of sympatric speciation in plants. The Sphagnum subsecundum complex includes six species in North America. Three have haploid gametophytes, and three are thought to have diploid gametophytes. Microsatellite analyses indicated that some plants of S. inundatum and S. lescurii are heterozygous at most loci, but others have only one allele at each locus. Flow cytometry and Feulgen staining showed that heterozygous plants have twice the genome size as plants with one allele per locus; thus, microsatellite patterns can be used to survey the distribution and abundance of haploid and diploid gametophytes. Microsatellite analyses also revealed that S. carolinianum is consistently diploid, but S. lescurii and S. inundatum include both haploid and diploid populations. The frequency of diploid plants in S. lescurii increases with latitude. In an analysis of one population of S. lescurii, both cytotypes co-occurred but were genetically differentiated with no evidence of interbreeding. The degree of genetic differentiation showed that the diploids were not derived from simple genome duplication of the local haploids. Heterozygosity appears to be fixed or nearly so in diploids, strongly suggesting that although morphologically indistinguishable from the haploids, they are derived by allopolyploidy. PMID- 21628168 TI - The age and biogeography of Citrus and the orange subfamily (Rutaceae: Aurantioideae) in Australasia and New Caledonia. AB - The geological history of Australasia, New Caledonia, and Southeast Asia, has been complex, resulting in competing biogeographic hypotheses for taxa found here. Alternative hypotheses-Gondwanan vicariance, rafting terranes, long distance dispersal-may be distinguished by different predicted divergence times between disjunct sister taxa. Taxa within Rutaceae subfamily Aurantioideae are ideal for testing these hypotheses because of their distributions. Therefore, the ages of Rutaceae and Aurantioideae were estimated using molecular dating. One data set comprised 51 sequences of rbcL and atpB with sampling across rosids and three fossil calibrations: crown Fabales+Fagales+Rosales (>94 Ma), Fabaceae (>51 Ma) and stem Ailanthus, Simaroubaceae (>52 Ma). Another data set comprised 81 Aurantioideae using >8 kb of chloroplast sequence and secondary calibration. Confidence in estimated divergence times was explored by varying the root age, dating method (strict, local, and relaxed clocks), and inclusion of internal calibrations. We conclude that the Rutaceae crown diverged in the Eocene (36.4 56.8 Ma, mean 47.6), whereas the Aurantioideae crown originated in the early Miocene (12.1-28.2 Ma, mean 19.8). This young age suggests that Gondwanan vicariance does not explain the distributions of extant Aurantioideae. Taxa found in New Caledonia may have arrived by separate transoceanic dispersal events. PMID- 21628169 TI - Genetic diversity and geographical structure of the pitcher plant Nepenthes vieillardii in New Caledonia: A chloroplast DNA haplotype analysis. AB - Among the many species that grow in New Caledonia, the pitcher plant Nepenthes vieillardii (Nepenthaceae) has a high degree of morphological variation. In this study, we present the patterns of genetic differentiation of pitcher plant populations based on chloroplast DNA haplotype analysis using the sequences of five spacers. We analyzed 294 samples from 16 populations covering the entire range of the species, using 4660 bp of sequence. Our analysis identified 17 haplotypes, including one that is widely distributed across the islands, as well as regional and private haplotypes. The greatest haplotype diversity was detected on the eastern coast of the largest island and included several private haplotypes, while haplotype diversity was low in the southern plains region. The parsimony network analysis of the 17 haplotypes suggested that the genetic divergence is the result of long-term isolation of individual populations. Results from a spatial analysis of molecular variance and a cluster analysis suggest that the plants once covered the entire serpentine area of New Caledonia and that subsequent regional fragmentation resulted in the isolation of each population and significantly restricted seed flow. This isolation may have been an important factor in the development of the morphological and genetic variation among pitcher plants in New Caledonia. PMID- 21628170 TI - The sporophyte-less filmy fern of eastern North America Trichomanes intricatum (Hymenophyllaceae) has the chloroplast genome of an Asian species. AB - Trichomanes intricatum, the sporophyte-less filmy fern of the eastern United States, has been considered to be a species whose sporophyte generation has become extinct or is possibly still present among the many species of Trichomanes s.l. in the new world tropics but unable to grow in a temperate climate. A close relationship to Asian species has heretofore not been considered. Comparison of rbcL and rps4-trnS sequences to species of Trichomanes s.l. reveals that T. intricatum shares its chloroplast genome with Crepidomanes schmidtianum of eastern Asia. Because C. schmidtianum is a sterile triploid and the ploidy level of T. intricatum is unknown, several scenarios leading to their sharing of these maternally inherited genes must be explored. PMID- 21628171 TI - Geographic divergence in floral morphology and scent in Linanthus dichotomus (Polemoniaceae). AB - Floral reproductive morphology and scent are of primary importance to pollinators in guiding foraging decisions. We compared the floral scent and reproductive morphology between two subspecies of Linanthus dichotomus (Polemoniaceae) that are taxonomically distinguished by geography and flowering time: the vespertine L. dichotomus subsp. dichotomus and the diurnal L. dichotomus subsp. meridianus. Disparity in flowering time between the two subspecies is accompanied by differences in flower visitors. We collected floral volatiles using dynamic headspace methods and analyzed them using gas chromatography and mass spectroscopy. Together, the subspecies produced a total of 39 floral scent compounds. Subspecies differ in the quantitative pattern of volatiles that attract noctuid moths (e.g., lilac aldehydes) vs. a more general suite of visitors (e.g., phenylacetaldehyde), but not in overall scent emission rates. A discriminant function analysis correctly distinguished between the two subspecies based on scent samples 86% of the time. We measured seven reproductive morphological traits; a discriminant function analysis distinguished between the two subspecies based on morphological samples 81% of the time. We found significant differences between subspecies in scent but not in individual morphological traits. The evidence presented here is most consistent with a hypothesis of pollinator-mediated selection. PMID- 21628172 TI - Foreword from the Editor-in-Chief1. PMID- 21628173 TI - Introduction to the Darwin special issue: The abominable mystery1. PMID- 21628174 TI - The meaning of Darwin's 'abominable mystery'. AB - Charles Darwin's "abominable mystery" has come to symbolize just about all aspects of the origin and early evolution of flowering plants. Yet, there has never been an analysis of precisely what Darwin thought was so abominably mysterious. Here I explicate Darwin's thoughts and frustrations with the fossil record of flowering plants as revealed in correspondence with Joseph Hooker, Gaston de Saporta, and Oswald Heer between 1875 and 1881. I also examine the essay by John Ball that prompted Darwin to write his "abominable mystery" letter to Hooker in July of 1879. Contrary to what is generally believed, Darwin's abominable mystery has little if anything to do with the fossil prehistory of angiosperms, identification of the closest relatives of flowering plants, questions of the homologies (and character transformations) of defining features of flowering plants, or the phylogeny of flowering plants themselves. Darwin's abominable mystery and his abiding interest in the radiation of angiosperms were never driven primarily by a need to understand the literal text of the evolutionary history of flowering plants. Rather, Darwin was deeply bothered by what he perceived to be an abrupt origin and highly accelerated rate of diversification of flowering plants in the mid-Cretaceous. This led Darwin to create speculative arguments for a long, gradual, and undiscovered pre-Cretaceous history of flowering plants on a lost island or continent. Darwin also took refuge in the possibility that a rapid diversification of flowering plants in the mid-Cretaceous might, if real, have a biological explanation involving coevolutionary interactions between pollinating insects and angiosperms. Nevertheless, although generations of plant biologists have seized upon Darwin's abominable mystery as a metaphor for their struggle to understand angiosperm history, the evidence strongly suggests that the abominable mystery is not about angiosperms per se. On the contrary, Darwin's abominable mystery is about his abhorrence that evolution could be both rapid and potentially even saltational. Throughout the last years of his life, it just so happens that flowering plants, among all groups of organisms, presented Darwin with the most extreme exception to his strongly held notion natura non facit saltum, nature does not make a leap. PMID- 21628175 TI - Reconstructing the ancestral angiosperm flower and its initial specializations. AB - Increasingly robust understanding of angiosperm phylogeny allows more secure reconstruction of the flower in the most recent common ancestor of extant angiosperms and its early evolution. The surprising emergence of several extant and fossil taxa with simple flowers near the base of the angiosperms Chloranthaceae, Ceratophyllum, Hydatellaceae, and the Early Cretaceous fossil Archaefructus (the last three are water plants)-has brought a new twist to this problem. We evaluate early floral evolution in angiosperms by parsimony optimization of morphological characters on phylogenetic trees derived from morphological and molecular data. Our analyses imply that Ceratophyllum may be related to Chloranthaceae, and Archaefructus to either Hydatellaceae or Ceratophyllum. Inferred ancestral features include more than two whorls (or series) of tepals and stamens, stamens with protruding adaxial or lateral pollen sacs, several free, ascidiate carpels closed by secretion, extended stigma, extragynoecial compitum, and one or several ventral pendent ovule(s). The ancestral state in other characters is equivocal: e.g., bisexual vs. unisexual flowers, whorled vs. spiral floral phyllotaxis, presence vs. absence of tepal differentiation, anatropous vs. orthotropous ovules. Our results indicate that the simple flowers of the newly recognized basal groups are reduced rather than primitively simple. PMID- 21628176 TI - Nonflowers near the base of extant angiosperms? Spatiotemporal arrangement of organs in reproductive units of Hydatellaceae and its bearing on the origin of the flower. AB - Reproductive units (RUs) of Trithuria, the sole genus of the early-divergent angiosperm family Hydatellaceae, are compared with flowers of their close relatives in Cabombaceae (Nymphaeales). Trithuria RUs combine features of flowers and inflorescences. They differ from typical flowers in possessing an "inside out" morphology, with carpels surrounding stamens; furthermore, carpels develop centrifugally, in contrast to centripetal or simultaneous development in typical flowers. Trithuria RUs could be interpreted as pseudanthia of two or more cymose partial inflorescences enclosed within an involucre, but the bractlike involucral phyllomes do not subtend partial inflorescences and hence collectively resemble a typical perianth. Teratological forms of T. submersa indicate a tendency to fasciation and demonstrate that the inside-out structure-the primary feature that separates RUs of Hydatellaceae from more orthodox angiosperm flowers-can be at least partially modified, thus producing a morphology that is closer to an orthodox flower. The Trithuria RU could be described as a "nonflower", i.e., a structure that contains typical angiosperm carpels and stamens but does not allow recognition of a typical angiosperm flower. The term nonflower could combine cases of secondary loss of flower identity and cases of a prefloral condition, similar to those that gave rise to the angiosperm flower. Nonhomology among some angiosperm flowers could be due to iterative shifts between nonfloral construction and flower/inflorescence organization of reproductive organs. Potential testing of these hypotheses using evolutionary-developmental genetics is explored using preliminary data from immunolocalization of the floral meristem identity gene LEAFY in T. submersa, which indicated protein expression at different hierarchical levels. PMID- 21628177 TI - More is better: the uses of developmental genetic data to reconstruct perianth evolution. AB - The origin and evolution of the perianth remains enigmatic. While it seems likely that an undifferentiated perianth consisting of tepals arose early in angiosperm evolution, it is unclear when and how differentiated perianths consisting of distinct organs, such as petals and sepals, arose. Phylogenetic reconstructions of ancestral perianth states across angiosperms have traditionally relied on morphological data from extant species, but these analyses often produce equivocal results. Here we describe the use of developmental genetic data as an additional strategy to infer the ancestral perianth character state for different angiosperm clades. By assessing functional data in combination with expression data in a maximum likelihood framework, we provide a novel approach for investigating the evolutionary history of the perianth. Results of this analysis provide new insights into perianth evolution and provide a proof of concept for using this strategy to explore the incorporation of developmental genetic data in character state reconstructions. As the assumptions outlined here are tested and more genetic data are generated, we hope that ancestral state reconstructions based on multiple lines of evidence will converge. PMID- 21628178 TI - One size fits all? Molecular evidence for a commonly inherited petal identity program in Ranunculales. AB - Petaloid organs are a major component of the floral diversity observed across nearly all major clades of angiosperms. The variable morphology and development of these organs has led to the hypothesis that they are not homologous but, rather, have evolved multiple times. A particularly notable example of petal diversity, and potential homoplasy, is found within the order Ranunculales, exemplified by families such as Ranunculaceae, Berberidaceae, and Papaveraceae. To investigate the molecular basis of petal identity in Ranunculales, we used a combination of molecular phylogenetics and gene expression analysis to characterize APETALA3 (AP3) and PISTILLATA (PI) homologs from a total of 13 representative genera of the order. One of the most striking results of this study is that expression of orthologs of a single AP3 lineage is consistently petal-specific across both Ranunculaceae and Berberidaceae. We conclude from this finding that these supposedly homoplastic petals in fact share a developmental genetic program that appears to have been present in the common ancestor of the two families. We discuss the implications of this type of molecular data for long held typological definitions of petals and, more broadly, the evolution of petaloid organs across the angiosperms. PMID- 21628179 TI - Floral variation and floral genetics in basal angiosperms. AB - Recent advances in phylogeny reconstruction and floral genetics set the stage for new investigations of the origin and diversification of the flower. We review the current state of angiosperm phylogeny, with an emphasis on basal lineages. With the surprising inclusion of Hydatellaceae with Nymphaeales, recent studies support the topology of Amborella sister to all other extant angiosperms, with Nymphaeales and then Austrobaileyales as subsequent sisters to all remaining angiosperms. Notable modifications from most recent analyses are the sister relationships of Chloranthaceae with the magnoliids and of Ceratophyllaceae with eudicots. We review "trends" in floral morphology and contrast historical, intuitive interpretations with explicit character-state reconstructions using molecular-based trees, focusing on (1) the size, number, and organization of floral organs; (2) the evolution of the perianth; (3) floral symmetry; and (4) floral synorganization. We provide summaries of those genes known to affect floral features that contribute to much of floral diversity. Although most floral genes have not been investigated outside of a few model systems, sufficient information is emerging to identify candidate genes for testing specific hypotheses in nonmodel plants. We conclude with a set of evo-devo case studies in which floral genetics have been linked to variation in floral morphology. PMID- 21628180 TI - Reconstructing the ancestral female gametophyte of angiosperms: Insights from Amborella and other ancient lineages of flowering plants. AB - For more than a century, the common ancestor of flowering plants was thought to have had a seven-celled, eight-nucleate Polygonum-type female gametophyte. It is now evident that not one, but in fact three, patterns of female gametophyte development and mature structure characterize the common ancestors of the four most ancient clades of extant angiosperms: Amborella-type, Nuphar/Schisandra-type and Polygonum-type. The Amborella-type female gametophyte is restricted to a single extant species, Amborella trichopoda, and at maturity consists of eight cells and nine nuclei. Development of the Amborella-type gametophyte is essentially identical to the Polygonum-type except that there is an additional and asynchronous cell division at the micropylar pole prior to maturation that produces a third synergid and the egg cell. The Nuphar/Schisandra-type female gametophyte is four-nucleate and four-celled and at maturity contains a typical three-celled egg apparatus and a central cell with a single haploid polar nucleus. This type of gametophyte appears to be universal among extant members of the Nymphaeales (including Hydatellaceae) and Austrobaileyales. Based on explicit reconstruction of character distribution and evolution, the Polygonum-type female gametophyte is certain to be representative of the common ancestors of monocots, eudicots, magnoliids, Ceratophyllaceae, and Chloranthaceae. There are compelling biological reasons to suggest that the four-celled, four-nucleate female gametophyte (as found in Nymphaeales and Austrobaileyales) is ancestral among angiosperms, with transitions to Polygonum-type female gametophytes separately in the Amborellales and in the ancient angiosperm clade that includes all angiosperms except Amborella, Nymphaeales, and Austrobaileyales. Subsequent to the evolution of a seven-celled, eight-nucleate Polygonum-type female gametophyte in the Amborellales, we hypothesize that a peramorphic increase in egg apparatus cell number took place and led to the unique situation in which there are three synergids in Amborella trichopoda. PMID- 21628181 TI - Amborella trichopoda (Amborellaceae) and the evolutionary developmental origins of the angiosperm progamic phase. AB - A remarkable number of the defining features of flowering plants are expressed during the life history stage between pollination and fertilization. Hand pollinations of Amborella trichopoda (Amborellaceae) in New Caledonia show that when the stigma is first receptive, the female gametophyte is near maturity. Pollen germinates within 2 h, and pollen tubes with callose walls and plugs grow entirely within secretions from stigma to stylar canal and ovarian cavity. Pollen tubes enter the micropyle within 14 h, and double fertilization occurs within 24 h. Hundreds of pollen tubes grow to the base of the stigma, but few enter the open stylar canal. New data from Amborella, combined with a review of fertilization biology of other early-divergent angiosperms, show that an evolutionary transition from slow reproduction to rapid reproduction occurred early in angiosperm history. I identify increased pollen tube growth rates within novel secretory carpel tissues as the primary mechanism for such a shift. The opportunity for prezygotic selection through interactions with the stigma is also an important innovation. Pollen tube wall construction and substantial modifications of the ovule and its associated structures greatly facilitated a new kind of reproductive biology. PMID- 21628182 TI - Pollination biology of basal angiosperms (ANITA grade). AB - The first three branches of the angiosperm phylogenetic tree consist of eight families with ~201 species of plants (the ANITA grade). The oldest flower fossil for the group is dated to the Early Cretaceous (115-125 Mya) and identified to the Nymphaeales. The flowers of extant plants in the ANITA grade are small, and pollen is the edible reward (rarely nectar or starch bodies). Unlike many gymnosperms that secrete "pollination drops," ANITA-grade members examined thus far have a dry-type stigma. Copious secretions of stigmatic fluid are restricted to the Nymphaeales, but this is not nectar. Floral odors, floral thermogenesis (a resource), and colored tepals attract insects in deceit-based pollination syndromes throughout the first three branches of the phylogenetic tree. Self incompatibility and an extragynoecial compitum occur in some species in the Austrobaileyales. Flies are primary pollinators in six families (10 genera). Beetles are pollinators in five families varying in importance as primary (exclusive) to secondary vectors of pollen. Bees are major pollinators only in the Nymphaeaceae. It is hypothesized that large flowers in Nymphaeaceae are the result of the interaction of heat, floral odors, and colored tepals to trap insects to increase fitness. PMID- 21628183 TI - Transmitting tissue architecture in basal-relictual angiosperms: Implications for transmitting tissue origins. AB - Carpel transmitting tissue is a major floral innovation that is essential for angiosperm success. It facilitates the rapid adhesion, hydration, and growth of the male gametophyte to the female gametophyte. As well, it functions as a molecular screen to promote male gametophytic competition and species-specific recognition and compatibility. Here, we characterize the transmitting tissue extracellular matrix (ECM) and pollen tube growth in basal-relictual angiosperms and test the hypothesis that a freely flowing ECM (wet stigma) was ancestral to a cuticle-bound ECM (dry stigma). We demonstrate that the most recent common ancestor of extant angiosperms produced an ECM that was structurally and functionally equivalent to a dry stigma. Dry stigmas are composed of a cuticle and primary wall that contains compounds that facilitate the adhesion and growth of the male gametophyte. These compounds include methyl-esterified homogalacturonans, arabinogalactan-proteins, and lipids. We propose that transmitting tissue evolved in concert with an increase in cuticle permeability that resulted from modifications in the biosynthesis and secretion of fatty acids needed for cuticle construction. Increased cuticle permeability exposed the male gametophyte to pre-existing molecules that enabled rapid male gametophyte adhesion, hydration, and growth as well as species-specific recognition and compatibility. PMID- 21628184 TI - Xylem of early angiosperms: Nuphar (Nymphaeaceae) has novel tracheid microstructure1. AB - SEM studies of xylem of stems of Nuphar reveal a novel feature, not previously reported for any angiosperm. Pit membranes of tracheid end walls are composed of coarse fibrils, densest on the distal (outside surface, facing the pit of an adjacent cell) surface of the pit membrane of a tracheid, thinner, and disposed at various levels on the lumen side of a pit membrane. The fibrils tend to be randomly oriented on the distal face of the pit membrane; the innermost fibrils facing the lumen take the form of longitudinally oriented strands. Where most abundantly present, the fibrils tend to be disposed in a spongiform, three dimensional pattern. Pores that interconnect tracheids are present within the fibrillar meshwork. Pit membranes on lateral walls of stem tracheids bear variously diminished versions of this pattern. Pits of root tracheids are unlike those of stems in that the lumen side of pit membranes bears a reticulum revealed on the outer surface of the tracheid after most of the thickness of a pit membrane is shaved away by the sectioning process. No fibrillar texturing is visible on the root tracheid pits when they are viewed from the inside of a tracheid. Tracheid end walls of roots do contain pores of various sizes in pit membranes. These root and stem patterns were seen in six species representing the two sections of Nuphar, plus one intersectional hybrid, as well as in one collection of Nymphaea, included for purposes of comparison. Differences between root and stem tracheids with respect to microstructure are consistent in all species studied. Microstructural patterns reported here for stem tracheid pits of Nymphaeaceae are not like those of Chloranthaceae, Illiciaceae, or other basal angiosperms. They are not referable to any of the patterns reported for early vascular plants. The adaptational nature of the pit membrane structure in these tracheids is not apparent; microstructure of pit membranes in basal angiosperms is more diverse than thought prior to study with SEM. PMID- 21628185 TI - Different gymnosperm outgroups have (mostly) congruent signal regarding the root of flowering plant phylogeny. AB - We examined multiple plastid genes from a diversity of gymnosperm lineages to explore the consistency of signal among different outgroups for rooting flowering plant phylogeny. For maximum parsimony (MP), most outgroups attach on a branch of the underlying ingroup tree that leads to Amborella. Maximum likelihood (ML) analyses either root angiosperms on a nearby branch or find split support for these neighboring root placements, depending on the outgroup. The inclusion of two species of Hydatellaceae, recently recognized as an ancient line of angiosperms, does not aid in inference of the root. Cost profiles for placing the root in suboptimal locations are highly correlated across most outgroup comparisons, even comparing MP and ML profiles. Those for Gnetales are the most deviant of all those considered. This divergent outgroup either attaches on a long eudicot branch with moderate bootstrap support in MP analyses or supports no particular root location in ML analysis. Removing the most rapidly evolving sites in rate classifications based on two divergent angiosperm root placements with Gnetales yields strongly conflicting root placements in MP analysis, despite substantial overlap in the estimated sets of conservative sites. However, the generally high consistency in rooting signal among distantly related gymnosperm clades suggests that the long branch connecting angiosperms to their extant relatives may not interfere substantially with inference of the angiosperm root. PMID- 21628186 TI - Phylogenetic relationships among seed plants: Persistent questions and the limits of molecular data. AB - Trees inferred from DNA sequence data provide only limited insight into the phylogeny of seed plants because the living lineages (cycads, Ginkgo, conifers, gnetophytes, and angiosperms) represent fewer than half of the major lineages that have been detected in the fossil record. Nevertheless, phylogenetic trees of living seed plants inferred from sequence data can provide a test of relationships inferred in analyses that include fossils. So far, however, significant uncertainty persists because nucleotide data support several conflicting hypotheses. It is likely that improved sampling of gymnosperm diversity in nucleotide data sets will help alleviate some of the analytical issues encountered in the estimation of seed plant phylogeny, providing a more definitive test of morphological trees. Still, rigorous morphological analyses will be required to answer certain fundamental questions, such as the identity of the angiosperm sister group and the rooting of crown seed plants. Moreover, it will be important to identify approaches for incorporating insights from data that may be accurate but less likely than sequence data to generate results supported by high bootstrap values. How best to weigh evidence and distinguish among hypotheses when some types of data give high support values and others do not remains an important problem. PMID- 21628187 TI - Seed ferns from the late Paleozoic and Mesozoic: Any angiosperm ancestors lurking there? AB - Five orders of late Paleozoic-Mesozoic seed ferns have, at one time or another, figured in discussions on the origin of angiosperms, even before the application of phylogenetic systematics. These are the Glossopteridales, Peltaspermales, Corystospermales, Caytoniales, and Petriellales. Although vegetative features have been used to suggest homologies, most discussion has focused on ovulate structures, which are generally interpreted as megasporophylls bearing seeds, with the seeds partially to almost completely enclosed by the megasporophyll (or cupule). Here we discuss current information about the reproductive parts of these plants. Since most specimens are impression-compression remains, homologizing the ovulate organs, deriving angiospermous homologues, and defining synapomorphies remain somewhat speculative. Although new specimens have increased the known diversity in these groups, a reconstruction of an entire plant is available only for the corystosperms, and thus hypotheses about phylogenetic position are of limited value. We conclude that, in the case of these seed plants, phylogenetic analysis techniques have surpassed the hard data needed to formulate meaningful phylogenetic hypotheses. Speculation on angiosperm origins and transitional stages in these fossils provides for interesting discussion, but currently it is still speculation, as the role of these groups in the origin of angiospermy continues to be cloaked in Darwin's mystery. PMID- 21628188 TI - Early Cretaceous mesofossils from Portugal and eastern North America related to the Bennettitales-Erdtmanithecales-Gnetales group. AB - Four new genera and six new species of fossil seed (Buarcospermum tetragonium, Lignierispermum maroneae, Lobospermum glabrum, L. rugosum, L. stampanonii, Rugonella trigonospermum) are described from five Early Cretaceous mesofossil floras from Portugal and eastern North America. The four genera are distinguished by differences in size, shape, and details of seed anatomy, but all are unusual in having an outer seed envelope with a distinctive anatomical structure that surrounds the nucellus and the integument. The integument is extended apically into a long, narrow micropylar tube. The four new genera are part of a diverse, but previously unrecognized, complex of extinct plants that was widespread in Early Cretaceous vegetation and that coexisted in similar habitats with early angiosperms. The distinctive structure of these seeds, and the strong similarities to other fossil seeds (Ephedra, Ephedripites, Erdtmanispermum, Raunsgaardispermum, and some Bennettitales) already known from the Early Cretaceous, suggests that this newly recognized complex of extinct plants, together with Bennettitales, Erdtmanithecales, and Gnetales (the BEG group), is phylogenetically closely related. PMID- 21628189 TI - Bennettitales from the Grisethorpe Bed (Middle Jurassic) at Cayton Bay, Yorkshire, UK. AB - Middle Jurassic fossil plants from the Grisethorpe Bed at Cayton Bay and Grisethorpe Bay, Yorkshire, UK, are preserved in a soft claystone, and plant mesofossils recovered by sieving reveal excellent details of external structure. Studies of these mesofossils complement previous work on macrofossils from the Grisethorpe Bed and allow the plant fossils in this classic flora to be studied in a similar way to those preserved in Cretaceous mesofloras. Bennettitales, a key group in discussions of how angiosperms may be related to other seed plants, are especially well represented among mesofossils from the Grisethorpe Bed. Abundant bennettitalean leaves, scale leaves, and fragments of pollen and ovulate organs provide new information on these extinct plants. In particular, a specimen of Williamsoniella coronata (presumed aborted) shows only weak differentiation between interseminal scales and ovules and provides further evidence of homology between these structures. PMID- 21628190 TI - Is the anthophyte hypothesis alive and well? New evidence from the reproductive structures of Bennettitales. AB - Bennettitales is an extinct group of seed plants with reproductive structures that are similar in some respects to both Gnetales and angiosperms, but systematic relationships among the three clades remain controversial. This study summarizes characters of bennettitalean plants and presents new evidence for the structure of cones and seeds that help clarify relationships of Bennettitales to flowering plants, Gnetales, and other potential angiosperm sister groups. Bennettitales have simple mono- or bisporangiate cones. Seeds are borne terminally on sporophylls. They have a unique structure that includes a nucellus with a solid apex, no pollen chamber, and a single integument, and they are clearly not enclosed by a cupule or other specialized structures. Such features differ substantially from Gnetales, flowering plants, and the seed fern Caytonia, providing no compelling evidence for the origin of the angiospermous carpel. Cladistic tests were performed to assess the strength of the "anthophyte hypothesis" and possible relationships of Bennettitales, Gnetales, and Caytonia to flowering plants. Our results do not support the anthophyte hypothesis for the origin of angiosperms by a transformation of fertile organs that were already aggregated into a cone or flower-like structure. However, the anthophyte topology of the seed plant tree continues to be supported by morphological analyses of living and extinct taxa. PMID- 21628191 TI - Distinguishing angiophytes from the earliest angiosperms: A Lower Cretaceous (Valanginian-Hauterivian) fruit-like reproductive structure. AB - A remarkably diverse Lower Cretaceous (Valanginian-Hauterivian) flora at Apple Bay, Vancouver Island, preserves seed plants at an important time of floristic evolutionary transition, about the same time as the earliest flowering plant megafossils. The fossils are permineralized in carbonate concretions and include tetrahedral seeds within cupule- or carpel-like structures. These enclosing structures, composed of elongate sclerenchyma cells with spiral thickenings that grade externally to a few layers of parenchyma, are vascularized by one collateral vascular bundle and lack trichomes. They apparently broke open to release the tightly enclosed seeds by valves. Seeds are similar to those of the Triassic seed fern Petriellaea, but are about 100 million years younger and differ in size, vascularization, integumentary anatomy, seed attachment, and number of seeds/cupule. These new seeds are described as Doylea tetrahedrasperma gen. et sp. nov., tentatively assigned to Corystospermales. Inverted cupules are reminiscent of an outer angiosperm integument rather than a carpel. Like fruits, cupules opened to release seeds at maturity, thereby foretelling several aspects of angiospermy. They show that nearly total ovule enclosure, a level of organization approaching angiospermy, was achieved by advanced seed ferns during the Mesozoic. PMID- 21628192 TI - Polyploidy and angiosperm diversification. AB - Polyploidy has long been recognized as a major force in angiosperm evolution. Recent genomic investigations not only indicate that polyploidy is ubiquitous among angiosperms, but also suggest several ancient genome-doubling events. These include ancient whole genome duplication (WGD) events in basal angiosperm lineages, as well as a proposed paleohexaploid event that may have occurred close to the eudicot divergence. However, there is currently no evidence for WGD in Amborella, the putative sister species to other extant angiosperms. The question is no longer "What proportion of angiosperms are polyploid?", but "How many episodes of polyploidy characterize any given lineage?" New algorithms provide promise that ancestral genomes can be reconstructed for deep divergences (e.g., it may be possible to reconstruct the ancestral eudicot or even the ancestral angiosperm genome). Comparisons of diversification rates suggest that genome doubling may have led to a dramatic increase in species richness in several angiosperm lineages, including Poaceae, Solanaceae, Fabaceae, and Brassicaceae. However, additional genomic studies are needed to pinpoint the exact phylogenetic placement of the ancient polyploidy events within these lineages and to determine when novel genes resulting from polyploidy have enabled adaptive radiations. PMID- 21628193 TI - Angiosperm diversification through time. AB - The extraordinary diversity of angiosperms is the ultimate outcome of the interplay of speciation and extinction, which determine the net diversification of different lineages. We document the temporal trends of angiosperm diversification rates during their early history. Absolute diversification rates were estimated for order-level clades using ages derived from relaxed molecular clock analyses that included or excluded a maximal constraint to angiosperm age. Diversification rates for angiosperms as a whole ranged from 0.0781 to 0.0909 net speciation events per million years, with dates from the constrained analysis. Diversification through time plots show an inverse relationship between clade age and rate, where the younger clades tend to have the highest rates. Angiosperm diversity is found to have mixed origins: slightly less than half of the living species belong to lineages with low to moderate diversification rates, which appeared between 130 and 102 Mya (Barremian-uppermost Albian; Lower Cretaceous). Slightly over half of the living species belong to lineages with moderate to high diversification rates, which appeared between 102 and 77 Mya (Cenomanian-mid Campanian; Upper Cretaceous). Terminal lineages leading to living angiosperm species, however, may have originated soon or long after the phylogenetic differentiation of the clade to which they belong. PMID- 21628194 TI - Darwin's second 'abominable mystery': Why are there so many angiosperm species? AB - The rapid diversification and ecological dominance of the flowering plants beg the question "Why are there so many angiosperm species and why are they so successful?" A number of equally plausible hypotheses have been advanced in response to this question, among which the most widely accepted highlights the mutually beneficial animal-plant relationships that are nowhere better developed nor more widespread than among angiosperm species and their biotic vectors for pollination and dispersal. Nevertheless, consensus acknowledges that there are many other attributes unique to or characteristic of the flowering plants. In addition, the remarkable coevolution of the angiosperms and pollination/dispersal animal agents could be an effect of the intrinsic adaptability of the flowering plants rather than a primary cause of their success, suggesting that the search for underlying causes should focus on an exploration of the genetic and epigenetic mechanisms that might facilitate adaptive evolution and speciation. Here, we explore angiosperm diversity promoting attributes in their general form and draw particular attention to those that, either individually or collectively, have been shown empirically to favor high speciation rates, low extinction rates, or broad ecological tolerances. Among these are the annual growth form, homeotic gene effects, asexual/sexual reproduction, a propensity for hybrid polyploidy, and apparent "resistance" to extinction. Our survey of the literature suggests that no single vegetative, reproductive, or ecological feature taken in isolation can account for the evolutionary success of the angiosperms. Rather, we believe that the answer to Darwin's second "abominable mystery" lies in a confluence of features that collectively make the angiosperms unique among the land plants. PMID- 21628195 TI - Anatomical variation in Cactaceae and relatives: Trait lability and evolutionary innovation. AB - The cacti have undergone extensive specialization in their evolutionary history, providing an excellent system in which to address large-scale questions of morphological and physiological adaptation. Recent molecular phylogenetic studies suggest that (1) Pereskia, the leafy genus long interpreted as the sister group of all other cacti, is likely paraphyletic, and (2) Cactaceae are nested within a paraphyletic Portulacaceae as a member of the "ACPT" clade (Anacampseroteae, Cactaceae, Portulaca, and Talinum). We collected new data on the vegetative anatomy of the ACPT clade and relatives to evaluate whether patterns in the distributions of traits may provide insight into early events in the evolutionary transition to the cactus life form. Many traits had high levels of homoplasy and were mostly equivocal with regard to infraclade relationships of ACPT, although several characters do lend further support to a paraphyletic Pereskia. These include a thick stem cuticle, prominent stem mucilage cells, and hypodermal calcium oxalate druses, all of which are likely to be important traits for stem water storage and photosynthesis. We hypothesize that high lability of many putative "precursor" traits may have been critical in generating the organismal context necessary for the evolution of an efficient and integrated photosynthetic stem. PMID- 21628196 TI - Morphological variation of intervessel pit membranes and implications to xylem function in angiosperms. AB - Pit membranes between xylem vessels have been suggested to have functional adaptive traits because of their influence on hydraulic resistance and vulnerability to embolism in plants. Observations of intervessel pit membranes in 26 hardwood species using electron microscopy showed significant variation in their structure, with a more than 25-fold difference in thickness (70-1892 nm) and observed maximum pore diameter (10-225 nm). In some SEM images, pit membrane porosity was affected by sample preparation, although pores were resolvable in intact pit membranes of many species. A significant relationship (r(2) = 0.7, P = 0.002) was found between pit membrane thickness and maximum pore diameter, indicating that the thinner membranes are usually more porous. In a subset of nine species, maximum pore diameter determined from SEM was correlated with pore diameter calculated from air-seeding thresholds (r(2) = 0.8, P < 0.001). Our data suggest that SEM images of intact pit membranes underestimate the porosity of pit membranes in situ. Pit membrane porosity based on SEM offers a relative estimate of air-seeding thresholds, but absolute pore diameters must be treated with caution. The implications of variation in pit membrane thickness and porosity to plant function are discussed. PMID- 21628197 TI - Developmental disaster1: A novel mutation causing defects during vegetative and inflorescence development in maize (Zea mays, Poaceae). AB - Axillary meristems, which give rise to branches and flowers, play a critical role in plant architecture and reproduction. To understand how axillary meristems initiate, we have screened for mutants with defects in axillary meristem initiation to uncover the genes controlling this process. These mutants, called the barren class of mutants in maize (Zea mays), have defects in axillary meristem initiation during both vegetative and reproductive development. Here, we identify and characterize a new member of the barren class of mutants named Developmental disaster1 (Dvd1), due to the pleiotropic effects of the mutation. Similar to the barren mutants, Dvd1 mutants have fewer branches, spikelets, florets, and floral organs in the inflorescence due to defects in the initiation of axillary meristems. Furthermore, double mutant analysis with teosinte branched1 shows that dvd1 also functions in axillary meristems during vegetative development. However, unlike the barren mutants, Dvd1 mutants are semidwarf due to the production of shorter internodes, and they produce leaves in the inflorescence due to the outgrowth of bract leaf primordia. The suite of defects seen in Dvd1 mutants, together with the genetic interaction of Dvd1 with barren inflorescence2, suggests that dvd1 is a novel regulator of axillary meristem and internode development. PMID- 21628198 TI - Ephemeral clonal integration in Calathea marantifolia (Marantaceae): Evidence of diminished integration over time. AB - A major advantage of clonal growth forms is the intergenerational transfer of resources through vascular connections (clonal integration). Connections linking ramets can be persistent or ephemeral. For species with ephemeral connections, whether the extent of clonal integration changes over time is unclear. To address this issue, we tracked water movement using an isotopic label and assessed the demographic performance of parent and offspring ramets over time in a severing experiment. Our study system was the understory herb Calathea marantifolia, which has parent ramets that produce vegetative bulbils (clonal offspring) that pass through distinct pre- and post-rooting stages. Little water was transported between parents and offspring, and the direction of movement was primarily from parent to pre-rooting offspring. Anatomical observations of inter-ramet connections showed that vascular bundles were twice as abundant in parent stems compared to inter-ramet connections. Severing inter-ramet connections reduced the growth of offspring ramets but not parents. Survival of pre-rooting offspring was reduced by 10% due to severing, but post-rooting offspring were not affected. Our results suggest that offspring ramets of C. marantifolia are weaned from their parent as they progress from pre- to post-rooting stages. PMID- 21628199 TI - Growth and survival across a gap-understory gradient: Contrast in performance of sexually vs. clonally produced offspring. AB - Sexually and clonally produced offspring may respond to environmental heterogeneity by growing and surviving at different rates. In forest understories, the availability of light ranges from low in shaded, closed canopy to high in tree-fall gaps. We experimentally investigated the growth and survival of both types of offspring in three treatments (gap centers, gap edges, and shaded understory) over 16 months. We expected the demographic performance of both types of offspring to be highest in the centers of gaps and lowest in the shaded understory. However, we expected seedlings to be more sensitive to the gradient in light (larger difference in growth and survival between light levels) than vegetative offspring because of their small size and lack of connection to maternal resources. Both offspring types grew fastest and obtained their largest sizes in gap centers. Contrary to our expectations, offspring types differed in which light conditions favored highest survival. Seedlings survived best in gap centers, while vegetative offspring had their highest survival in the shaded understory. In agreement with our hypothesis, survival and growth of seedlings were more sensitive to light availability, showing a large difference in growth and survival between light levels, compared to vegetative offspring. PMID- 21628200 TI - Diversity and structure of landraces of Agave grown for spirits under traditional agriculture: A comparison with wild populations of A. angustifolia (Agavaceae) and commercial plantations of A. tequilana. AB - Traditional farming communities frequently maintain high levels of agrobiodiversity, so understanding their agricultural practices is a priority for biodiversity conservation. The cultural origin of agave spirits (mezcals) from west-central Mexico is in the southern part of the state of Jalisco where traditional farmers cultivate more than 20 landraces of Agave angustifolia Haw. in agroecosystems that include in situ management of wild populations. These systems, rooted in a 9000-year-old tradition of using agaves as food in Mesoamerica, are endangered by the expansion of commercial monoculture plantations of the blue agave variety (A. tequilana Weber var. Azul), the only agave certified for sale as tequila, the best-known mezcal. Using intersimple sequence repeats and Bayesian estimators of diversity and structure, we found that A. angustifolia traditional landraces had a genetic diversity (H(BT) = 0.442) similar to its wild populations (H(BT) = 0.428) and a higher genetic structure ((B) = 0.405; (B) =0. 212). In contrast, the genetic diversity in the blue agave commercial system (H(B) = 0.118) was 73% lower. Changes to agave spirits certification laws to allow the conservation of current genetic, ecological and cultural diversity can play a key role in the preservation of the traditional agroecosystems. PMID- 21628201 TI - Consistency between marker- and genealogy-based heritability estimates in an experimental stand of Prosopis alba (Leguminosae). AB - Prosopis represents a valuable forest resource in arid and semiarid regions. Management of promising species requires information about genetic parameters, mainly the heritability (h(2)) of quantitative profitable traits. This parameter is traditionally estimated from progeny tests or half-sib analysis conducted in experimental stands. Such an approach estimates h(2) from the ratio of between family/total phenotypic variance. These analyses are difficult to apply to natural populations of species with a long life cycle, overlapping generations, and a mixed mating system, without genealogical information. A promising alternative is the use of molecular marker information to infer relatedness between individuals and to estimate h(2) from the regression of phenotypic similarity on inferred relatedness. In the current study we compared h(2) of 13 quantitative traits estimated by these two methods in an experimental stand of P. alba, where genealogical information was available. We inferred pairwise relatedness by Ritland's method using six microsatellite loci. Relatedness and heritability estimates from molecular information were highly correlated to the values obtained from genealogical data. Although Ritland's method yields lower h(2) estimates and tends to overestimate genetic correlations between traits, this approach is useful to predict the expected relative gain of different quantitative traits under selection without genealogical information. PMID- 21628202 TI - Earliest orchid macrofossils: Early Miocene Dendrobium and Earina (Orchidaceae: Epidendroideae) from New Zealand. AB - Fossil leaves of two Early Miocene orchids (Dendrobium and Earina) are reported from New Zealand. The distinctive, raised tetra- to cyclocytic stomatal subsidiary cells of Earina and characteristic papilla-like absorbing glands and "ringed" guard cells of Dendrobium support the placement of the fossils into these genera. These therefore represent the first Orchidaceae macrofossils with cuticular preservation, the oldest records for subfamily Epidendroideae, as well as the first New Zealand and southern hemisphere fossil records for Orchidaceae. These taxa belong in basal clades to the Vandeae/Cymbideae or Epidendreae (Earina) and the Australasian clade of Dendrobium sensu lato. This phylogenetic placement demonstrates expansion of epiphytic orchids into Zealandia by the mid Cenozoic and an important role for southern continents in the diversification of Orchidaceae. PMID- 21628203 TI - Ontogeny and ecology of the filicalean fern Oligocarpia gothanii (Gleicheniaceae) from the Middle Permian of China. AB - Reinvestigation of Oligocarpia gothanii Halle (Gleicheniaceae) from the Permian Period of China has uncovered a rare demonstration of ontogenetic succession in numerous intact plants and has led to emendation of the diagnosis and typification of the species. Reconstruction of the fern's vegetative life cycle shows a small rosette-shaped juvenile plant with immature fronds of pinnae undifferentiated into pinnules. The second stage is a series of leafy shoots increasing in size and complexity and bearing fronds essentially comparable with those of the adult plants. Mature plants have sphenopteroid-type fronds differentiated into vegetative and fertile fronds. Close association of plants on the same bedding plane and the presence of a connecting root network between shoots, show that juvenile plants spread by vegetative propagation using underground stolons. Sedimentological information and co-occurrence with the bryophyte Thallites hallei Lundblad suggests that O. gothanii occupied a wetland habitat and was preserved in an obrution event consistent with flooding of marginal areas in a fluvio-deltaic setting. Although recent studies on Oligocarpia have focused exclusively on reproductive features, new ontogenetic information suggests that plants growing in rapidly changing environments may have been reliant on vegetative propagation and would not have needed to reach sexual maturity for successful reproduction. PMID- 21628204 TI - Three extant genera of freshwater thalassiosiroid diatoms from Middle Eocene sediments in northern Canada. AB - The evolutionary history of diatoms is only constrained partially by the fossil record. The timing of several key events, such as initial colonization of freshwater habitats by marine taxa, remains poorly resolved. Numerous specimens of the genera Cyclotella, Discostella, and Puncticulata (Ochrophyta: Thalassiosirales) have been recovered in Middle Eocene lacustrine sediments from the Giraffe Pipe locality in the Northwest Territories, Canada. These diatoms extend the fossil record of the family Stephanodiscaceae to at least 40 million years before present (Ma) and thus provide a new evolutionary milepost for the thalassiosiroid diatoms, an important clade of centric diatoms with global representation in both marine and freshwater environments. The quality of the fossil material enables detailed investigations of areolae, fultoportulae, and rimoportulae, revealing direct morphological affinities with a number of extant taxa. These observations extend the antiquity of several characters of phylogenetic importance within the thalassiosiroid diatoms, including the fultoportula, and imply that the entire lineage is considerably older than prior constraints from the fossil record, as suggested independently by several recent molecular phylogenies. PMID- 21628205 TI - Can feral weeds evolve from cultivated radish (Raphanus sativus, Brassicaceae)? AB - Cultivated plants that cannot survive on their own often have maladaptive domestication traits. Unharvested crop seeds may generate feral populations, at times causing serious weed problems, but little is known about the evolution of ferality. We explored the potential for cultivated radish, Raphanus sativus, to become feral, given that closely related taxa (e.g., R. raphanistrum and crop wild hybrids) are well-documented weeds. First, we measured the population growth of five experimental, cultivated, self-seeding radish populations in Michigan, USA, for three generations. Three late-flowering populations went extinct, and two others apparently hybridized with local R. raphanistrum. A common garden experiment showed that the two surviving populations had earlier flowering, smaller root diameters, and greater individual fecundity than did nonhybridized populations. We also used artificial selection to measure the evolutionary potential for earlier flowering. After two generations of strong selection, two of three lineages flowered earlier and produced more seeds than control lineages, but insufficient genetic variation prevented dramatic evolution of crop phenotypes. In summary, it seems unlikely that radishes could spontaneously become feral in our study area without gene flow from R. raphanistrum. Applying these approaches to other cultivated species may provide a better understanding of mechanisms promoting the evolution of feral weeds. PMID- 21628206 TI - Is Oligomeris (Resedaceae) indigenous to North America? Molecular evidence for a natural colonization from the Old World. AB - Oligomeris linifolia constitutes one of the few examples of intercontinental disjunctions at the species level between the arid regions of the Old World and SW North America. The status of the American populations has been obscure, with some authors considering the populations to be introduced, whereas others believe them to be native. To clarify these conflicting opinions, we performed phylogeographic analyses using nuclear ribosomal ITS and plastid trnL-F and rps16 sequences to infer the origin of the disjunct American populations. Two independent molecular clock approaches based on ITS and cpDNA sequences (rbcL, matK, trnL-F) were used to estimate a divergence time of O. linifolia. Low levels of sequence divergence and estimates of relatively recent splits of Oligomeris lineages disagree with the vicariance hypotheses traditionally suggested to account for New-Old World disjunctions. In addition, significant genetic differentiation of American populations does not indicate a recent anthropogenic introduction. Morphological uniformity and the sharing of haplotypes between disjunct populations, together with the molecular clock results, suggest that a long-distance dispersal event from the Old Word to SW North America may have taken place during the Quaternary, in spite of limited dispersal mechanisms in Oligomeris. PMID- 21628207 TI - Familial placement and relations of Rehmannia and Triaenophora (Scrophulariaceae s.l.) inferred from five gene regions. AB - Accurate classification systems based on evolution are imperative for biological investigations. The recent explosion of molecular phylogenetics has resulted in a much improved classification of angiosperms. More than five phylogenetic lineages have been recognized from Scrophulariaceae sensu lato since the family was determined to be polyphyletic; however, questions remain about the genera that have not been assigned to one of the segregate families of Scrophulariaceae s.l. Rehmannia Liboschitz and Triaenophora Solereder are such genera with uncertain familial placement. There also is debate whether Triaenophora should be segregated from Rehmannia. To evaluate the phylogenetic relations between Rehmannia and Triaenophora, to find their closest relatives, and to verify their familial placement, we conducted phylogenetic analyses of the sequences of one nuclear DNA (ITS) region and four chloroplast DNA gene regions (trnL-F, rps16, rbcL, and rps2) individually and combined. The analyses showed that Rehmannia and Triaenophora are each strongly supported as monophyletic and together are sister to Orobanchaceae. This relation was corroborated by phytochemical and morphological data. Based on these data, we suggest that Rehmannia and Triaenophora represent the second nonparasitic branch sister to the remainder of Orobanchaceae (including Lindenbergia). PMID- 21628208 TI - Predicting the allometry of leaf surface area and dry mass. AB - The manner in which increases in leaf surface area S scale with respect to increases in leaf dry mass M(t) within and across species has important implications to understanding the ability of plants to harvest sunlight, grow, and ultimately reproduce. Thus far, no mechanistic explanation has been advanced to explain why prior work shows that the scaling exponent governing the S to M(t) relationship is generally significantly less than one (i.e., S ? M(t)(alpha < 1.0)) such that increases in M(t) yield diminishing returns with respect to increases in S across most species. Here, we show analytically why this phenomenon occurs and present equations that predict trends observed in the numerical values of scaling exponents for the S vs. M(t) relationships observed across dicot tree species and two aquatic vascular plant species. PMID- 21628209 TI - The adaptive value of cued seed dispersal in desert plants: Seed retention and release in Mammillaria pectinifera (Cactaceae), a small globose cactus. AB - Serotiny, or delayed seed dispersal, is common in fluctuating environments because it hedges the risks of establishment. Mammillaria pectinifera (Cactaceae) facultatively expels fruits in the year they are produced or retains them to disperse the seed over several years. We tested whether M. pectinifera increased fruit expulsion as a response to increased rainfall. While no fruit expulsion was observed in 1997, a dry year, in the wetter 1998 around 20% of all fruits formed were expelled from the maternal plant. A greenhouse experiment showed that high moisture results in the plants expelling all their fruits. Because in 1998 establishment was five times higher than in 1997, this response seems to be highly adaptive: Active fruit expulsion and consequent seed release increases the probability of establishment during pulses of high precipitation. PMID- 21628210 TI - Limitations within "The Limits to Tree Height"1. AB - Koch et al. (Nature 428: 851-854) measured various parameters that were thought to limit the height of Sequoia sempervirens from northern California and concluded that the maximum height for this species is 122-130 m because within this range: (1) Irreversible embolism formation was proposed to occur when the xylem pressure was less than -1.9 MPa. (2) The leaf mass to area ratio exponentially approached 833 g*m(-2). (3) The discrimination against (13)CO(2) exponentially approached -20. (4) Light-saturated photosynthesis per unit leaf mass decreased to zero, indicating no net gain in leaf biomass. These conclusions are questioned here by reassessing the assumed limits to the biophysical parameters and by reexamining the proposed linear and exponential relationships between these parameters and tree height. It is concluded that: (1) Embolism repair mechanisms could have occurred at -2.7 MPa. (2) The leaf mass to area ratio could be a result of, rather than a determinant of, the large differential between cellular turgor and the xylem pressure. (3) The discrimination against (13)CO(2) may show two populations of foliage with apparent linear relationships with height rather than one exponential relationship. (4) The light-saturated photosynthesis per unit leaf mass as a measure of biomass investment in leaf expansion excludes investment in branch and trunk wood. As a result, tree height may be limited by a long-term balance between dieback and continued growth. PMID- 21628211 TI - A response to: Limitations within "The Limits to Tree Height". AB - Here we respond to the communication in American Journal of Botany (96: 542-544 in this issue) by Netting, who proposes several ways in which our paper "The Limits to Tree Height" (Nature 428: 851-854) may have erred in estimating the biophysical limits to height growth in Sequoia sempervirens. We first explain that because embolism repair requires long time periods and is generally incomplete, xylem vulnerability characteristics offer a sound basis for estimating performance limits in woody plants. We reaffirm our earlier use of vertical gradients of foliar carbon isotope composition with new data for S. sempervirens. We support these arguments with reference to studies in other tree species. We take exception with Netting's view that the turgor pressure-cell expansion relationship for Zea mays is applicable to S. sempervirens. Finally, we second Netting's call for more work on carbon allocation vis a vis height growth limits. PMID- 21628212 TI - Diversification of inflorescence development in the PCK clade (Poaceae: Panicoideae: Paniceae). AB - In grasses, inflorescence diversification and its correlation with species evolution are intriguing and not well understood. Part of this problem lies in our lack of comprehension about the inflorescence morphological complexity of grasses. We focused our study on the PCK clade (named for phosphoenol pyruvate carboxykinase), a well-supported monophyletic group for which the relationships among its taxa are not well resolved. Interestingly, the PCK clade has an extensive diversity of adult inflorescence forms. A comparative developmental approach can help us to understand the basis of such morphological differences as well as provide characters that can be used in phylogenetic studies of the group. Using SEM studies, we demonstrate that inflorescence morphology in this clade is even more complex than what is typically observed in adult forms. We describe a number of new characters, and some classical features previously used for taxonomic purposes are redefined on the basis of development. We also define four morphological groups combining adult inflorescence form and development, and we discuss some of the evolutionary aspects of inflorescence diversification in the PCK clade. Taxonomic delimitation among genera in the PCK clade remains confusing and unclear where molecular and morphological studies support different classifications. PMID- 21628213 TI - Embryo sac, endosperm, and seed of Nemophila (Boraginaceae) relative to taxonomy, with a remark on embryogeny in Pholistoma. AB - Studies on embryology and seed morphology are complementary to molecular phylogenetics and of special value at the genus level. This paper discusses the delimitation and evolutionary relationships of genera within the tribe Hydrophylleae of the Boraginaceae. The seven Nemophila species characterized by a conspicuous seed appendage are similar in embryology and seed structure. The ovule is tenuinucellate and unitegmic with a meristematic tapetum. The embryo sac penetrating the nucellar apex is of the Polygonum type, has short-lived antipodal cells, and an embryo sac haustorium. The endosperm is cellular, producing two terminal endosperm haustoria, of which the chalazal has a lateral branch. Embryogeny is of the Chenopodiad type (as in Pholistoma). The seed coat is formed from the small-celled inner epidermis of the integument. The large-celled outer epidermis of the integument disintegrates into scattered cells. Seed pits evolve from irregularly placed inner epidermal cells of the integument. The chalazal part of the ovule produces a cucullus, that functions as an ant-attracting elaiosome. Those species of Nemophila with a conspicuous cucullus form a natural genus. Nemophila is most closely related to Pholistoma. The integumentary seed pits of Nemophila might have evolved from ovular seed pits similar to those in Pholistoma. PMID- 21628214 TI - Early floral development of Heliconia latispatha (Heliconiaceae), a key taxon for understanding the evolution of flower development in the Zingiberales. AB - We present new comparative data on early floral development of Heliconia latispatha, an ecologically and horticulturally important tropical plant within the order Zingiberales. Modification of the six members of two androecial whorls is characteristic of Zingiberales, with a reduction in number of fertile stamen from five or six in the banana families (Musaceae, Strelitziaceae, Lowiaceae, and Heliconiaceae) to one in Costaceae and Zingiberaceae and one-half in Marantaceae and Cannaceae. The remaining five infertile stamens in these later four families (the ginger families) are petaloid, and in Costaceae and Zingiberaceae fuse together to form a novel structure, the labellum. Within this developmental sequence, Heliconiaceae share with the ginger families the possession of an antisepalous staminode, a synapomorphy that has been used to place Heliconiaceae as sister to the ginger family clade. Here, we use epi-illumination light microscopy and reconstruction of serial sections to investigate the ontogeny of the Heliconia flower with emphasis on the ontogeny of the staminode. We compare floral development in Heliconia with that previously described for other species of Zingiberales. A comparison of floral structure and development across Zingiberales is presented to better understand the evolution of the flower in this charismatic group of tropical plants. PMID- 21628215 TI - Fern-spore-feeder interaction in temperate forests in Japan: Sporing phenology and spore-feeding insect community. AB - Angiosperms are widely appreciated to have flowering schedules, but far less attention has been paid to the timing of spore production by ferns. Although a range of abiotic factors are likely responsible for the timing of fern sporing, spore predation by specialist spore-feeding insects may also exert selective forces on timing. As a step toward understanding ecological factors that affect the evolution of fern sporing phenology, we tracked annual sporing patterns and examined associations with spore-feeding insects in 38 ferns species in two Japanese temperate forests. Most sporing occurred during June through August, the period of highest temperature and precipitation at the study sites, but some species produced spores during a limited period in spring or very late autumn. Over 70% of all species examined were attacked by spore-feeders, which consisted of seven polyphagous and five oligophagous species. Spore feeders occurred predominantly during June through September, and stathmopodid moth larvae consumed up to 70% of the mature sporangia. Thus, although the warm and humid conditions in the summer is likely favored for prothallial growth and fertilization, spring or late-autumn sporing in some species may have evolved as an adaptation to escape spore predation by spore feeders. PMID- 21628216 TI - Drought response in self-compatible species of tomato (Solanaceae). AB - Wild tomatoes occur in habitats from the extremely dry Atacama Desert to moist areas in the Andean highlands, which may have resulted in adaptation of populations or species to differences in soil moisture availability. However, when two accessions representing extremes in habitat water availability from each of the five self-compatible species were grown in a common garden, we observed no differences in leaf physiological responses to soil drought within or between species. All five species had drought avoidance characteristics with the same threshold soil moisture availability for decline of assimilation, stomatal conductance, and leaf water potential (Psi(l)) in response to slowly decreasing soil moisture. After rewatering, all species rapidly recovered to near predrought Psi(l), but bulk leaf solute potential after recovery did not indicate any osmotic adjustment. The lack of variation in shoot physiological traits during soil drought is unexpected as water deficit is commonly thought to have imposed selective pressure in the evolution of plant physiology. However, species did differ in assimilation under nonstressed conditions, which may contribute to differential soil water conservation and growth or survival during drought. PMID- 21628217 TI - Should structure-function relations be considered separately for homobaric vs. heterobaric leaves? AB - Tree and shrub species can be differentiated into two major groups based on their substantially different leaf anatomy: heterobaric and homobaric. In contrast to homobaric leaves, heterobaric leaves have bundle sheath extensions (BSEs) that create transparent regions on their lamina. Recent studies have shown that BSEs transfer visible light to internal mesophyll layers, thus affecting the photosynthetic performance of heterobaric leaves. Whether the two leaf types also differ in other functional and structural traits has not been addressed, nor have any structure-function relations. Here, we measured key anatomical and physiological parameters and tested their relationships in 30 species with different leaf types. Heterobaric leaves were thinner with lower leaf mass per area, had higher nitrogen concentration per mass, were (13)C-enriched, and achieved comparable photosynthetic capacity per area but had higher photosynthetic capacity per mass compared to homobaric leaves. Relations between leaf construction cost, nitrogen concentration, and photosynthesis followed the general pattern of the "leaf economic spectrum," but differed between homobaric and heterobaric leaves. We suggest that the mechanisms controlling these relations differ between the two leaf types, presumably due to their distinct anatomy. PMID- 21628218 TI - Increasing demands on limited water resources: Consequences for two endangered plants in Amargosa Valley, USA. AB - Recent population expansion throughout the Southwest United States has created an unprecedented demand for already limited water resources, which may have severe consequences on the persistence of some species. Two such species are the federally protected Nitrophila mohavensis (Chenopodiaceae) and Grindelia fraxino pratensis (Asteraceae) found in Amargosa Valley, one valley east of Death Valley, California. Because both species are federally protected, no plant material could be harvested for analysis. We therefore used a chamber system to collect transpired water for isotopic analysis. After a correction for isotopic enrichment during transpiration, delta(18)O values of plant xylem water were significantly different between N. mohavensis and G. fraxino-pratensis throughout the study. Using a multisource mixing model, we found that both N. mohavensis and G. fraxino-pratensis used soil moisture near the soil surface in early spring when surface water was present. However, during the dry summer months, G. fraxino pratensis tracked soil moisture to deeper depths, whereas N. mohavensis continued to use soil moisture near the soil surface. These results indicate that pumping groundwater and subsequently lowering the water table may directly prevent G. fraxino-pratensis from accessing water, whereas these same conditions may indirectly affect N. mohavensis by reducing surface soil moisture and thus its ability to access water. PMID- 21628219 TI - Efficient translation in chloroplasts requires element(s) upstream of the putative ribosome binding site from atpI. AB - Thousands of proteins make up a chloroplast, but fewer than 100 are encoded by the chloroplast genome. Despite this low number, expression of chloroplast encoded genes is essential for plant survival. Every chloroplast has its own gene expression system with a major regulatory point at the initiation of protein synthesis (translation). In chloroplasts, most protein-encoding genes contain elements resembling the ribosome binding sites (RBS) found in prokaryotes. In vitro, these putative chloroplast ribosome binding sequences vary in their ability to support translation. Here we report results from an investigation into effects of the predicted RBS for the tobacco chloroplast atpI gene on translation in vivo. Two reporter constructs, differing only in their 5'-untranslated regions (5'UTRs) were stably incorporated into tobacco chloroplast genomes and their expression analyzed. One 5'UTR was derived from the wild-type (WT) atpI gene. The second, Holo-substitution (Holo-sub), had nonchloroplast sequence replacing all wild-type nucleotides, except for the putative RBS. The abundance of reporter RNA was the same for both 5'UTRs. However, translation controlled by Holo-sub was less than 4% that controlled by WT. These in vivo experiments support the idea that translation initiation in land plant chloroplasts depends on 5'UTR elements outside the putative RBS. PMID- 21628220 TI - Anatomy and development of fruits of Lauraceae from the Middle Eocene Princeton Chert. AB - Investigations of the Middle Eocene Princeton Chert reveal evidence for the connection of lauraceous flowers to fruits through a developmental series. Youngest fruits are found with attached floral remnants. Later stages show receptacle enlargement, fruit wall thickening, and the development of abundant sclereid clusters. Mature fruits are borne on a shallow receptacle and have an endocarp palisade layer of radially elongate cells with stellate outlines, an inner mesocarp layer of radiately arranged sclereid clusters, and a fleshy outer mesocarp layer containing numerous idioblasts with contents. Each mature fruit bears a single seed retaining the outer integument with an innermost radially elongate transfusion cell layer. Mature seeds contain a cellular embryo bearing idioblasts. Fruits are distinguishable from previously described anatomically preserved fossil taxa. This study represents the only documented developmental reconstruction of fossil fruits of Lauraceae and that self-pruning evolved prior to the Eocene. Anatomical modifications over the developmental sequence indicate that different stages of maturity preserved together, may be erroneously identified as several taxa at a fossil locality. Fossil morphotypes typically underestimate species number, but this study suggests that the number of inferred species based on fruit types may be inflated for Lauraceae, potentially exaggerating the tropical interpretation of the paleoenvironment. PMID- 21628221 TI - Gravity control of growth form in Brassica rapa and Arabidopsis thaliana (Brassicaceae): Consequences for secondary metabolism. AB - How gravity influences the growth form and flavor components of plants is of interest to the space program because plants could be used for food and life support during prolonged missions away from the planet, where that constant feature of Earth's environment does not prevail. We used plant growth hardware from prior experiments on the space shuttle to grow Brassica rapa and Arabidopsis thaliana plants during 16-d or 11-d hypergravity treatments on large-diameter centrifuge rotors. Both species showed radical changes in growth form, becoming more prostrate with increasing g-loads (2-g and 4-g). In Brassica, height decreased and stems thickened in a linear relationship with increasing g-load. Glucosinolates, secondary compounds that contribute flavor to Brassica, decreased by 140% over the range of micro to 4-g, while the structural secondary compound, lignin, remained constant at ~15% (w/w) cell wall dry mass. Stem thickening at 4 g was associated with substantial increases in cell size (47%, 226%, and 33% for pith, cortex, and vascular tissue), rather than any change in cell number. The results, which demonstrate the profound effect of gravity on plant growth form and secondary metabolism, are discussed in the context of similar thigmostresses such as touch and wind. PMID- 21628222 TI - Population genetic structure of an endangered Utah endemic, Astragalus ampullarioides (Fabaceae). AB - The endangered Shivwits milkvetch, Astragalus ampullarioides, is a perennial, herbaceous plant. This Utah endemic was federally listed as endangered in 2001 because of its high habitat specificity and low numbers of individuals and populations. All habitat currently occupied by A. ampullarioides was designated as critical by the U.S. Fish and Wildlife Service in 2006 as a result of conservation litigation. We used AFLP markers to assess genetic differentiation among the seven extant populations and quantified genetic diversity in each. Six different AFLP markers resulted in 217 unambiguous polymorphic loci. We used multiple methods to examine any changes in population genetic structure in this species over time. Results indicate that A. ampullarioides had much higher gene flow among populations in the past, but has since fragmented into regional genetic units. These regions further fragmented genetically, and extant populations have differentiated through genetic drift. Populations had low levels of gene flow, even between geographically close populations. Rapid urban development reduces gene flow among regions and encroaches on populations of A. ampullarioides and remaining patches of unoccupied habitat. The genetic makeup of each of the extant populations should be carefully considered in management decisions such as population establishment or augmentation. PMID- 21628223 TI - A molecular phylogeny of the orange subfamily(Rutaceae: Aurantioideae) using nine cpDNA sequences. AB - The breeding of new, high-quality citrus cultivars depends on dependable information about the relationships of taxa within the tribe Citreae; therefore, it is important to have a well-supported phylogeny of the relationships between species not only to advance breeding strategies, but also to advance conservation strategies for the wild taxa. The recent history of the systematics of Citrus (Rutaceae: Aurantioideae) and its allies, in the context of Rutaceae taxonomy as a whole, is reviewed. The most recent classification is tested using nine cpDNA sequence regions in representatives of all genera of the subfam. Aurantioideae (save Limnocitrus) and numerous species and hybrids referred to Citrus s.l. Aurantioideae are confirmed as monophyletic. Within Aurantioideae, tribe Clauseneae are not monophyletic unless Murraya s.s. and Merrillia are removed to Aurantieae. Within tribe Aurantieae, the three traditionally recognized subtribes are not monophyletic. Triphasiinae is not monophyletic unless Oxanthera is returned to Citrus (Citrinae). Balsamocitrinae is polyphyletic. Feroniella, traditionally considered allied closely to Limonia (=Feronia), is shown to be nested in Citrus. The proposed congenericity of Severinia and Atalantia is confirmed. The most recent circumscription of Citrus is strongly supported by this analysis, with hybrids appearing with their putative maternal parents. The genus was resolved into two clades, one comprising wild species from New Guinea, Australia, and New Caledonia (formerly Clymenia, Eremocitrus, Microcitrus, Oxanthera), but surprisingly also Citrus medica, traditionally believed to be native in India. The second clade is largely from the Asian mainland (including species formerly referred to Fortunella and Poncirus). PMID- 21628224 TI - Phylogeny and classification of the species-rich pantropical showy genus Ixora (Rubiaceae-Ixoreae) with indications of geographical monophyletic units and hybrids. AB - Species-rich genera often have various conflicting circumscriptions from independent regional flora treatments. Testing the monophyly of these groups of plants is an important step toward the establishment of a phylogenetic classification. The genus Ixora of the tribe Ixoreae in the subfamily Ixoroideae (coffee family or Rubiaceae) is a species-rich pantropical genus of ca. 500 species. Phylogenetic analyses of Ixoreae based on combined sequence data from one nuclear (nrETS) and two chloroplast (rps16 and trnT-F) markers reveal the paraphyly of Ixora as presently delimited and also show that the tribe can be subdivided into three major clades: the Mascarene/neotropical/Malagasy/African clade, the Pacific clade, and the Asian clade. Given the lack of morphological synapomorphies supporting the different Ixora clades and the morphological consistency of the ingroup taxa, we propose a broad circumscription of Ixora including all its satellite genera: Captaincookia, Doricera, Hitoa, Myonima, Sideroxyloides, Thouarsiora, and Versteegia. The current infrageneric classification of Ixora is not supported. The different Ixora subclades represent geographical units. Nuclear and chloroplast tree topologies were partially incongruent, indicating at least four potential natural hybridization events. Other conflicting positions for the cultivated species are most likely due to anthropogenic hybridization. PMID- 21628225 TI - Chloroplast diversity in a putative hybrid swarm of Ponderosae (Pinaceae). AB - The Ponderosae subsection of the genus Pinus contains numerous taxa in disjunct mountain ranges of southern Arizona and New Mexico, differing for several leaf and cone traits, key among which is the number of leaf needles per fascicle. Trees with three needles are often found together with trees having five needles and mixed numbers. One taxonomic hypothesis is that there are swarms of hybrids between P. ponderosa and P. arizonica. A second hypothesis is that there are spatial mixtures of two separate taxa, five-needle P. arizonica and a "taxon X" containing three needle and mixed needle trees. We genotyped chloroplasts in one putative hybrid swarm on Mt. Lemmon using microsatellite markers and show that cpDNA is almost completely differentiated between two separate morphotypes corresponding to P. arizonica and "taxon X." Little if any introgression has occurred on Mt. Lemmon, and the simplest explanation is that little or no effective hybridization has occurred. Further results indicate that not only is taxon X not of hybrid origin, it is more closely related to nonregional Ponderosae other than P. ponderosa and P. arizonica. The results further suggest that other putative hybrid swarms in the region are also spatial mixtures of distinct taxa. PMID- 21628226 TI - Decaploidy in Fragaria iturupensis (Rosaceae). AB - The strawberry genus, Fragaria (Rosaceae), has a base chromosome number of x = 7. Cultivated strawberries (F. *ananassa nothosubsp. ananassa) are octoploid (2n = 8x = 56) and first hybridized from F. chiloensis subsp. chiloensis forma chiloensis * F. virginiana subsp. virginiana. Europe has no known native octoploid species, and only one Asian octoploid species has been reported: F. iturupensis, from Iturup Island. Our objective was to examine the chromosomes of F. iturupensis. Ploidy levels of wild strawberry species, include diploid (2n = 2x = 14), tetraploid (2n = 4x = 28), pentaploid (2n = 5x = 35), hexaploid (2n = 6x = 42), octoploid (2n = 8x = 56), and nonaploid (2n = 9x = 63). Artificial triploid (2n = 3x = 21), tetraploid, pentaploid, octoploid, decaploid (2n = 10x = 70), 16-ploid, and 32-ploid plants have been constructed and cultivated. Surprisingly, chromosome counts and flow cytometry revealed that F. iturupensis includes natural decaploid genotypes with 2n = 10x = 70 chromosomes. This report is the first of a naturally occurring decaploid strawberry species. Further research on F. iturupensis and exploration on northern Pacific islands is warranted to ascertain the phylogeny and development of American octoploid species. PMID- 21628227 TI - Gelatinous fibers are widespread in coiling tendrils and twining vines. AB - Although the coiling of tendrils and the twining of vines has been investigated since Darwin's time, a full understanding of the mechanism(s) of this coiling and twining ability has not yet been obtained. In a previous study (Planta 225: 485 498), gelatinous (G) fibers in tendrils of redvine occurred concomitantly with the ability to coil, strongly indicating their role in the coiling process. In this study, tendrils and twining vines of a number of species were examined using microscopic and immunocytochemical techniques to determine if a similar presence and distribution of these fibers exists in other plant species. Tendrils that coiled in many different directions had a cylinder of cortical G fibers, similar to redvine. However, tendrils that coiled only in a single direction had gelatinous fibers only along the inner surface of the coil. In tendrils with adhesive tips, the gelatinous fibers occurred in the central/core region of the tendril. Coiling occurred later in development in these tendrils, after the adhesive pad had attached. In twining stems, G fibers were not observed during the rapid circumnutation stage, but were found at later stages when the vine's position was fixed, generally one or two nodes below the node still circumnutating. The number and extent of fiber development correlated roughly with the amount of torsion required for the vine to ascend a support. In contrast, species that use adventitious roots for climbing or were trailing/scrambling-type vines did not have G fibers. These data strongly support the concept that coiling and twining in vines is caused by the presence of G fibers. PMID- 21628228 TI - Morphological and genetic variations of Potentilla matsumurae (Rosaceae) between fellfield and snowbed populations. AB - Identifying ecological factors associated with local differentiation of populations is important for understanding microevolutionary processes. Alpine environments offer a unique opportunity to investigate the effects of habitat specific selective forces and gene flow limitations among populations at a microscale on local adaptation because the heterogeneous snowmelt patterns in alpine ecosystems provide steep environmental changes. We investigated the variation in morphological traits and enzyme loci between fellfield and snowbed populations of Potentilla matsumurae, a common alpine herb with a wide distribution along snowmelt gradients in northern Japan. We found significant differences in morphological traits between fellfield and snowbed habitats in a northern distribution region. These differences were maintained when plants were grown under uniform conditions in a greenhouse. Allozyme variations among 15 populations from geographically separated regions with different historical backgrounds showed that the populations are more genetically differentiated between the fellfield and snowbed habitats within a region than between populations occupying the same habitat type in different regions. These results suggest that variation in snowmelt regimes could be a driving force creating local adaptation and genetic differentiation of alpine plant populations. PMID- 21628229 TI - Ecology of leaf teeth: A multi-site analysis from an Australian subtropical rainforest. AB - Teeth are conspicuous features of many leaves. The percentage of species in a flora with toothed leaves varies inversely with temperature, but other ecological controls are less known. This gap is critical because leaf teeth may be influenced by water availability and growth potential and because fossil tooth characters are widely used to reconstruct paleoclimate. Here, we test whether ecological attributes related to disturbance, water availability, and growth strategy influence the distribution of toothed species at 227 sites from Australian subtropical rainforest. Both the percentage and abundance of toothed species decline continuously from riparian to ridge-top habitats in our most spatially resolved sample, a result not related to phylogenetic correlation of traits. Riparian lianas are generally untoothed and thus do not contribute to the trend, and there is little association between toothed riparian species and ecological attributes indicating early successional lifestyle and disturbance response. Instead, the pattern is best explained by differences in water availability. Toothed species' proportional richness declines with proximity to the coast, also a likely effect of water availability because salt stress causes physiological drought. Our study highlights water availability as an important factor impacting the distribution of toothed species across landscapes, with significance for paleoclimate reconstructions. PMID- 21628230 TI - A comparative study of oak (Quercus, Fagaceae) seedling physiology during summer drought in southern California. AB - Natural recruitment of oaks appears to be declining throughout the northern hemisphere. Summer drought poses a potentially important barrier to oak recruitment in southern California. To evaluate this barrier, we grew evergreen Quercus agrifolia and deciduous Q. lobata from seeds near parental trees. We measured water relations, chlorophyll fluorescence, and gas exchange during these seedlings' fourth and fifth summers and compared them to neighboring adults. Most seedlings had substantially lower values for predawn xylem pressure potential (Psi(pd)), minimum photosystem II (PSII) quantum efficiency (Phi(PSIIMIN)), maximum quantum efficiency for PSII under dark-adapted leaf conditions (Fv/Fm), and maximum photosynthetic assimilation (Amax), and higher values for maximum nonphotochemical quenching (NPQmax) than did conspecific adults. The high, unvarying Psi(pd) values of the adults suggest they use perennially available groundwater. Quercus lobata seedlings commonly had lower values for Psi(pd) than did Q. agrifolia, and values for Psi(pd) and Phi(PSIIMIN) were significantly related to size in Q. lobata but not in Q. agrifolia. These data suggest important interspecific differences in root architecture. Lower values for Phi(PSIIMIN), Fv/Fm, and higher NPQmax in Q. agrifolia indicate that Q. agrifolia seedlings were usually under more stress than Q. lobata, which typically had higher Amax rates than did Q. agrifolia seedlings. Diurnal photosynthesis curves were quite flat for Q. agrifolia, but they peaked in the morning for Q. lobata. Established seedlings appeared to be under more stress than adults, but this stress did not appear severe enough to cause death. Access to perennially available groundwater may be crucial for the seedling to sapling transition. PMID- 21628231 TI - Effects of polyploidy on secondary chemistry, physiology, and performance of native and invasive genotypes of Solidago gigantea (Asteraceae). AB - The role of polyploidy in facilitating invasiveness of introduced plants has not been well explored. Examination of traits of diploid and polyploid plants in both their native and introduced ranges can shed light on evolutionary processes occurring postintroduction in invasive plants. We determined the distribution and prevalence of cytotypes of Solidago gigantea in both its native range (USA) and introduced range (Europe), and measured a suite of biochemical, physiological, and reproductive characters for plants from both continents. Tetraploids were the most frequent cytotype encountered on both continents, while hexaploids were found only in the USA. Hexaploids were the most distinctive cytotype, with fewer differences observed between diploids and tetraploids. Comparison of diploids and tetraploids in the USA and Europe showed that traits changed in concert for both cytotypes. Both diploids and tetraploids in Europe had reduced concentrations of three classes of secondary chemical and invested relatively more into rhizomes than into flowers. The same changes occurring in both cytotypes in the introduced range show that altered phenotypes of European plants are not due to shifts in the proportions of cytotypes but instead occur within them. There was no evidence that polyploids evolve more quickly in the introduced range. PMID- 21628232 TI - Population genetic structure of native versus naturalized sympatric shrub willows (Salix; Salicaceae). AB - Vegetative propagation of an introduced species can contribute significantly to its ability to spread and become naturalized, potentially in competition with native species. This study focused on the naturalization of a willow shrub, Salix purpurea, which was introduced to the United States from Europe and is commonly sympatric with the native shrub willow, S. eriocephala. Both species are capable of vegetative and sexual reproduction, but little is known about their relative frequency, nor the impact of clonal propagation on population-level genetic diversity. We analyzed genotypes at several microsatellite loci in 993 individuals belonging to 30 subpopulations of S. eriocephala and 28 subpopulations of S. purpurea in areas of sympatry across three watersheds to compare their genetic diversity and genetic structure. Our results revealed six subpopulations of S. purpurea containing plants with identical multilocus genotypes, while clonal individuals were rare among S. eriocephala populations. These species are dioecious with relatively high levels of heterozygosity, but S. eriocephala had much higher allelic diversity and genotypic diversity than did S. purpurea. These results strongly suggest that vegetative propagation has contributed to the naturalization of S. purpurea and has resulted in higher levels of genetic differentiation among S. purpurea populations than among native S. eriocephala populations. PMID- 21628233 TI - Inflorescences contribute more than rosettes to lifetime carbon gain in Arabidopsis thaliana (Brassicaceae). AB - A metamorphosis from rosette to inflorescence in many annuals shifts photosynthetic tissue from a two-dimensional array in the soil boundary layer during cool months to a three-dimensional structure in the troposphere as spring progresses. We propose that this shift allows escape from both self-shading and an increasingly stressful boundary layer microclimate, permitting continued increases in growth. As a first step in exploring this hypothesis, we compared the lifetime C gain, water loss, and instantaneous water use efficiency (WUE) of five Arabidopsis thaliana genotypes by measuring gas exchange across the life cycle. On average, the inflorescence contributed 55% (+/- 5% SE) of lifetime C gain, but only 25% of lifetime water loss. Mean inflorescence WUE was nearly fourfold that of the rosette. The inflorescence continued to fix C after rosette senescence. The percentage inflorescence: total C gain varied among genotypes, from 36% to 93%. Genotypes differed in WUE for both structures. We suggest that local climates may have selected for divergence in these traits. For many annuals and winter annuals, understanding C and water budgets and their evolution must include measures of both rosette and inflorescence gas exchange. PMID- 21628234 TI - The evolution of floral variation without pollinator shifts in Gorteria diffusa (Asteraceae). AB - One of the most widely accepted explanations for floral diversification in angiosperms is the pollinator-shift model developed by Verne Grant and Ledyard Stebbins. According to this model, the most profound changes in floral traits (such as morphology, color, patterning and scent) occur when plants undergo adaptive shifts between pollinator classes. We tested this model through investigations of geographical variation in floral form and pollinator assemblages in the South African annual daisy Gorteria diffusa. This species has elaborate insect-like ornaments on the capitulum, which attract bee flies belonging to the genus Megapalpus. We found unprecedented levels of geographically structured intraspecific variation and identified 14 discrete forms that vary in the morphology and ornamentation of the capitulum. This variation is not due to phenotypic plasticity because differences among forms were maintained in plants grown from seed in a common garden experiment. Contrary to predictions from the pollinator-shift model, all populations, regardless of floral phenotype, were pollinated primarily by a single species of Megapalpus bee fly. Much of the extensive variation in floral form in G. diffusa therefore appears to have arisen without evolutionary shifts between pollinator types. PMID- 21628235 TI - The coexistence of bicellular and tricellular pollen in Annona cherimola (Annonaceae): Implications for pollen evolution. AB - Most angiosperms release bicellular pollen. However, in about one-third of extant angiosperms, the second pollen mitosis occurs before anthesis such that pollen is tricellular upon release. The shift from bicellular to tricellular development has occurred several times independently, but its causes are largely unknown. In this work, we observed the coexistence of both kinds of pollen at anther dehiscence in Annona cherimola, a species that belongs to the basal angiosperm family Annonaceae. Examination of pollen cell number during anther development showed that this coexistence was due to a late mitosis starting shortly before pollen shedding. Both types of pollen germinated equally well over the course of development. Because variable proportions of bicellular and tricellular pollen were observed at different sampling times, we tested the role of temperature by performing field and growth chamber experiments, which showed that higher temperatures near anthesis advanced the time of pollen mitosis II. The results show that selection could favor the production of tricellular pollen under certain environmental circumstances that prime rapid pollen germination and provide evidence of a system in which developmental variation persists, but that can be modified by external factors such as temperature. PMID- 21628236 TI - Interspecific pollinator movements reduce pollen deposition and seed production in Mimulus ringens (Phrymaceae). AB - Movement of pollinators between coflowering plant species may influence conspecific pollen deposition and seed set. Interspecific pollinator movements between native and showy invasive plants may be particularly detrimental to the pollination and reproductive success of native species. We explored the effects of invasive Lythrum salicaria on the reproductive success of Mimulus ringens, a wetland plant native to eastern North America. Pollinator flights between these species significantly reduced the amount of conspecific pollen deposited on Mimulus stigmas and the number of seeds in Mimulus fruits, suggesting that pollen loss is an important mechanism of competition for pollination. Although pollen loss is often attributed to pollen wastage on heterospecific floral structures, our novel findings suggest that grooming by bees as they forage on a competitor may also significantly reduce outcross pollen export and seed set in Mimulus ringens. PMID- 21628237 TI - Phylogeny of the tribe Indigofereae (Leguminosae-Papilionoideae): Geographically structured more in succulent-rich and temperate settings than in grass-rich environments. AB - This analysis goes beyond many phylogenies in exploring how phylogenetic structure imposed by morphology, ecology, and geography reveals useful evolutionary data. A comprehensive range of such diversity is evaluated within tribe Indigofereae and outgroups from sister tribes. A combined data set of 321 taxa (over one-third of the tribe) by 80 morphological characters, 833 aligned nuclear ribosomal ITS/5.8S sites, and an indel data set of 33 characters was subjected to parsimony analysis. Notable results include the Madagascan dry forest Disynstemon resolved as sister to tribe Indigofereae, and all species of the large genus Indigofera comprise just four main clades, each diagnosable by morphological synapomorphies and ecological and geographical predilections. These results suggest niche conservation (ecology) and dispersal limitation (geography) are important processes rendering signature shapes to the Indigofereae phylogeny in different biomes. Clades confined to temperate and succulent-rich biomes are more dispersal limited and have more geographical phylogenetic structure than those inhabiting tropical grass-rich vegetation. The African arid corridor, particularly the Namib center of endemism, harbors many of the oldest Indigofera lineages. A rates analysis of nucleotide substitutions confirms that the ages of the oldest crown clades are mostly younger than 16 Ma, implicating dispersal in explaining the worldwide distribution of the tribe. PMID- 21628238 TI - Investigating species boundaries in the Giliopsis group of Ipomopsis (Polemoniaceae): Strong discordance among molecular and morphological markers. AB - As a first step in elucidating mechanisms of speciation in the Giliopsis group of Ipomopsis (Polemoniaceae), we examined patterns of morphological and genetic differentiation and crossability. This group comprises three species that diverged very recently: two perennials, I. guttata and I. tenuifolia, and one annual, I. effusa. Analysis of phenotypic variation established that the three species are distinct for floral characters, and this differentiation is maintained in a locality containing both perennial species. Next, we assessed the genealogical relationships with AFLPs. All sampled individuals of I. effusa clustered together, a result in accord with its genetic isolation. The perennials, which retain interfertility, were not resolved as sister taxa. Rather, individuals sampled from the single I. guttata population that is sympatric with I. tenuifolia were genetically more similar to I. tenuifolia samples than they were to conspecifics. This pattern may be due to substantial introgression of I. tenuifolia genomic regions that do not contribute to floral phenotype in I. guttata. Our result adds to mounting evidence that plant species, as defined by morphological characters, are often not genomically cohesive. Taken together, our data warrant caution in delimiting species with genetic markers alone, and, importantly, suggest that selection on species-diagnostic morphological characters can be sufficiently strong to counteract extensive gene flow. PMID- 21628239 TI - In vivo study of developmental programmed cell death using the lace plant (Aponogeton madagascariensis; Aponogetonaceae) leaf model system. AB - Programmed cell death (PCD) is required for many morphological changes, but in plants it has been studied in much less detail than in animals. The unique structure and physiology of the lace plant (Aponogeton madagascariensis) is well suited for the in vivo study of developmental PCD. Live streaming video and quantitative analysis, coupled with transmission electron microscopy, were used to better understand the PCD sequence, with an emphasis on the chloroplasts. Dividing, dumbbell-shaped chloroplasts persisted until the late stages of PCD. However, the average size and number of chloroplasts, and the starch granules associated with them, declined steadily in a manner reminiscent of leaf senescence, but distinct from PCD described in the Zinnia tracheary element system. Remaining chloroplasts often formed a ring around the nucleus. Transvacuolar strands, which appeared to be associated with chloroplast transport, first increased and then decreased. Mitochondrial streaming ceased abruptly during the late stages of PCD, apparently due to tonoplast rupture. This rupture occurred shortly before the rapid degradation of the nucleus and plasma membrane collapse, in a manner also reminiscent of the Zinnia model. The presence of numerous objects in the vacuoles suggests increased macro-autophagy before cell death. These objects were rarely observed in cells not undergoing PCD. PMID- 21628240 TI - Scaling relations for a functionally two-dimensional plant: Chamaesyce setiloba (Euphorbiaceae). AB - Many characteristics of plants and animals scale with body size as described by allometric equations of the form Y = betaM(alpha), where Y is an attribute of the organism, beta is a coefficient that varies with attribute, M is a measure of organism size, and alpha is another constant, the scaling exponent. In current models, the frequently observed quarter-power scaling exponents are hypothesized to be due to fractal-like structures. However, not all plants or animals conform to the assumptions of these models. Therefore, they might be expected to have different scaling relations. We studied one such plant, Chamaesyce setiloba, a prostrate annual herb that grows to functionally fill a two-dimensional space. Number of leaves scaled slightly less than isometrically with total aboveground plant mass (alpha ~ 0.9) and substantially less than isometrically with dry total stem mass (alpha = 0.82), showing reduced allocation to leaf as opposed to stem tissue with increasing plant size. Additionally, scalings of the lengths and radii of parent and daughter branches differed from those predicted for three dimensional trees and shrubs. Unlike plants with typical three-dimensional architectures, C. setiloba has distinctive scaling relations associated with its particular prostrate herbaceous growth form. PMID- 21628241 TI - Crown structure and wood properties: Influence on tree sway and response to high winds. AB - Wind can alter plant growth and cause extensive, irreversible damage in forested areas. To better understand how to mitigate the effects of wind action, we investigated the sensitivity of tree aerodynamic behavior to the material and geometrical factors characterizing the aerial system. The mechanical response of a 35-yr-old maritime pine (Pinus pinaster, Pinaceae) submitted to static and dynamic wind loads is simulated with a finite element model. The branching structure is represented in three dimensions. Factor effects are evaluated using a fractional experimental design. Results show that material properties play only a limited role in tree dynamics. In contrast, small morphological variations can produce extreme behaviors such as either very little or nearly critical dissipation of stem oscillations. Slender trees are shown to be relatively more vulnerable to stem breakage than uprooting. Dynamic loading leads to deflections and forces up to 20% higher near the base of the tree than those calculated for a static loading of similar magnitude. Effects of branch geometry on dynamic amplification are substantial yet not linear. The flexibility of the aerial system is found to be critical to reducing the resistance to the airflow and thus to minimizing the risk of failure. PMID- 21628242 TI - Leaf herbivory increases floral fragrance in male but not female Cucurbita pepo subsp. texana (Cucurbitaceae) flowers. AB - Mutualisms are key interactions that affect population dynamics and structure communities, but the extent to which mutualists can attract potential partners may depend on community context. Many studies have shown that leaf herbivory reduces pollinator visitation and have focused on reduced floral visual display and rewards as potential mechanisms. However, olfactory display plays a critical role in mediating interactions between plants, herbivores, and pollinators. We simulated leaf damage in Cucurbita pepo subsp. texana and measured fragrance emission and other floral characters of both male and female flowers. Contrary to our expectations, damage increased fragrance production, but only in male flowers. Female flowers, which were bigger and produced more fragrance than males, were unaffected by leaf damage. The greatest increase in floral fragrance compounds was in the terpenoids, which we hypothesize could be byproducts of defensively induced cucurbitacins, or they may function defensively themselves. In summary, this study is the first to demonstrate changes in floral fragrance due to leaf damage. Such changes in floral fragrance following herbivory may be a critical and overlooked mechanism mediating interactions between plants, herbivores, and pollinators. PMID- 21628243 TI - Survival costs of reproduction in a short-lived perennial plant: Live hard, die young. AB - According to life-history theory, reproductive investments involve costs in terms of growth, future fecundity, and/or survival. However, studies to date have often failed to detect costs of reproduction, with survival costs among the less documented. We investigated the cost of reproduction in Helianthemum squamatum (Cistaceae), a short-lived perennial of semiarid Mediterranean environments. After experimental flower removal, we evaluated next season's growth, reproduction, and survival of the plants. We also monitored an indicator of plant physiological status (F(v)/F(m)) and leaf nutrient concentration at key phenological stages during reproduction. Survival rate in deblossomed plants was significantly higher than in control plants. As far as we know, this is the first experimental evidence of a survival cost of reproduction in a perennial plant. In contrast, no cost to growth or reproduction was found during the next season, and no significant differences in F(v)/F(m) or leaf nutrients were found between control and deblossomed plants. Helianthemum squamatum's success in semiarid Mediterranean ecosystems seems to rely on a persistent seed bank, combined with a sustained high reproductive output at the expense of survival. We conclude that this strategy might be more common than previously thought among short-lived shrubby plants growing in stressful Mediterranean areas. PMID- 21628244 TI - Effects of flowering plant density on pollinator visitation, pollen receipt, and seed production in Delphinium barbeyi (Ranunculaceae). AB - Variation in flowering plant density can have conflicting effects on pollination and seed production. Dense flower patches may attract more pollinators, but flowers in those patches may also compete for pollinator visits and abiotic resources. We examined how natural and experimental conspecific flowering plant density affected pollen receipt and seed production in a protandrous, bumble bee pollinated wildflower, Delphinium barbeyi (Ranunculaceae). We also compared floral sex ratios, pollinator visitation rates, and pollen limitation of seed set from early to late in the season to determine whether these factors mirrored seasonal changes in pollen receipt and seed production. Pollen receipt increased with natural flowering plant density, while seed production increased across lower densities and decreased across higher flower densities. Experimental manipulation of flowering plant density did not affect pollinator visitation rate, pollen receipt, or seed production. Although pollinator visitation rate increased 10-fold from early to late in the season, pollen receipt and seed set decreased over the season. Seed set was never pollen-limited. Thus, despite widespread effects of flowering plant density on plant reproduction in other species, the effects of conspecific flowering plant density on D. barbeyi pollination and seed production are minor. PMID- 21628245 TI - Insights into reticulate evolution in Machaerantherinae (Asteraceae: Astereae): 5S ribosomal RNA spacer variation, estimating support for incongruence, and constructing reticulate phylogenies. AB - Although reticulate evolution has been a frequent occurrence during the history of plants, determining how it has contributed to plant evolution will require analyzing many nuclear loci and developing effective analytical methods. The objective of this study was to make progress toward meeting these requirements in the evolutionarily complex subtribe Machaerantherinae. The 5S ribosomal RNA (rRNA) spacer was investigated to characterize its structure and variation. Analysis of the spacer supported relationships that were mostly the same as those supported by ITS and ETS sequence data. Two methods were used to estimate support for 11 occurrences of incongruence between 5S/ITS/ETS and cpDNA data sets. The five best-supported incongruences were proposed to have had reticulate evolutionary histories. For Arida blepharophylla, Xanthisma rhizomatum, and Pyrrocoma, 5S and ITS/ETS evidence supported the same or similar relationships, indicating that these two regions of the nuclear genome were descended from the same ancestor or from two closely related ancestors, with cpDNA coming from a more distantly related ancestor. The 5S and ITS/ETS evidence disagreed on the relationships of Arida riparia, suggesting that its ITS/ETS region came from one ancestor and its 5S region and cpDNA from a different ancestor. PMID- 21628246 TI - Pentapetalum trifasciculandricus gen. et sp. nov., a thealean fossil flower from the Raritan Formation, New Jersey, USA (Turonian, Late Cretaceous). AB - The study of fossil flowers in the last 25 years has greatly increased our understanding of angiosperm diversification. Following that tradition, we here describe a new fossil taxon from the Late Cretaceous of New Jersey, Pentapetalum trifasciculandricus Martinez-Millan, Crepet et Nixon gen. et sp. nov. It includes actinomorphic pentamerous flowers with quincuncial calyx, imbricate corolla, numerous stamens of markedly different heights, and a superior tricarpellate ovary, which are morphologically consistent with the flowers of the Theaceae s.l. and of members of the order Theales sensu Cronquist. Cladistic analyses including 45 extant taxa plus the fossil, 61 morphological characters, and different combinations of the molecular markers rbcL, matK, trnL-trnF, matR, and ITS support its inclusion in the order Ericales sensu APG. Comparison with extant taxa using traditional methods of identification suggests a relation with the Theaceae s.s. (Stewartia), but the phylogenetic analyses do not support this view. Instead, the phylogenetic analysis suggests some relation to the Ternstroemiaceae/Pentaphylacaceae (Theaceae s.l.), exemplifying the importance of evaluating identifications in a phylogenetic context. The description of Pentapetalum further adds to the ample diversity of Ericales in the Late Cretaceous. PMID- 21628247 TI - Spatial genetic structuring of baobab (Adansonia digitata, Malvaceae) in the traditional agroforestry systems of West Africa. AB - This study evaluates the spatial genetic structure of baobab (Adansonia digitata) populations from West African agroforestry systems at different geographical scales using AFLP fingerprints. Eleven populations from four countries (Benin, Ghana, Burkina Faso, and Senegal) had comparable levels of genetic diversity, although the two populations in the extreme west (Senegal) had less diversity. Pairwise F(ST) ranged from 0.02 to 0.28 and increased with geographic distance, even at a regional scale. Gene pools detected by Bayesian clustering seem to be a byproduct of the isolation-by-distance pattern rather than representing actual discrete entities. The organization of genetic diversity appears to result essentially from spatially restricted gene flow, with some influences of human seed exchange. Despite the potential for relatively long-distance pollen and seed dispersal by bats within populations, statistically significant spatial genetic structuring within populations (SGS) was detected and gave a mean indirect estimate of neighborhood size of ca. 45. This study demonstrated that relatively high levels of genetic structuring are present in baobab at both large and within population level, which was unexpected in regard to its dispersal by bats and the influence of human exchange of seeds. Implications of these results for the conservation of baobab populations are discussed. PMID- 21628248 TI - Phylogeographic patterns and demographic history of Schiedea globosa (Caryophyllaceae) on the Hawaiian Islands. AB - Geomorphological changes have been demonstrated to have had profound impacts on biodiversity, often leading to demographic expansions and contractions and allopatric divergence of taxa. We examined DNA sequence variation at two nuclear and one maternally inherited plastid locus among 10 populations of Schiedea globosa on the Hawaiian Islands to assess the primary factors shaping genetic structure, phylogeographic patterns, and the importance of geographic isolation to population divergence. Schiedea globosa has characteristics that may promote gene flow, including wind pollination and rafting of plants in ocean currents. However, we detected significant differentiation among populations on all islands except Hawaii, with the maternally inherited plastid locus having the greatest genetic structure (F(ST) = 0.81). Migration rates across all loci are less than one migrant per generation. We found evidence of growth in several populations and on the islands of Molokai and Maui, which supports population expansion associated with the formation of Maui Nui during the last glacial maximum. Similar to data for many other Hawaiian taxa, these data suggest S. globosa originated on Oahu and subsequently colonized Molokai, Maui, and Hawaii in progression. Given the high level of genetic structure, allopatric divergence will likely contribute to further divergence of populations. PMID- 21628249 TI - Size-dependent pollen:ovule ratios and the allometry of floral sex allocation in Clarkia (Onagraceae) taxa with contrasting mating systems. AB - Multiple field populations of two pairs of diploid sister taxa with contrasting mating systems in the genus Clarkia (Onagraceae) were surveyed to test predictions concerning the effects of resource status, estimated as plant size, on pollen and ovule production and on the pollen:ovule (P:O) ratio of flowers. Most theoretical models of size-dependent sex allocation predict that, in outcrossing populations, larger plants should allocate more resources to female function. Lower P:O ratios in larger plants compared to smaller plants have been interpreted as supporting this prediction. In contrast, we predicted that P:O ratio should not vary with plant size in predominantly selfing plants, in which each flower contributes to reproductive success equally through male and female function. We found that, in all four taxa, both ovule and pollen production per flower usually increased significantly with plant size and that the shape of this relationship was decelerating. However, ovule production either decelerated more rapidly than or at the same rate as pollen production with plant size. Consequently,the P:O ratio increased or had no relationship with plant size. This relationship was population-specific (not taxon-specific) and independent of the mating system. Possible explanations for the increasing maleness with plant size are discussed. PMID- 21628250 TI - Synthetic polyploids of Tragopogon miscellus and T. mirus (Asteraceae): 60 Years after Ownbey's discovery. AB - In plants, polyploidy has been a significant evolutionary force on both recent and ancient time scales. In 1950, Ownbey reported two newly formed Tragopogon allopolyploids in the northwestern United States. We have made the first synthetic lines of T. mirus and T. miscellus using T. dubius, T. porrifolius, and T. pratensis as parents and colchicine treatment of F(1) hybrids. We also produced allotetraploids between T. porrifolius and T. pratensis, which are not known from nature. We report on the crossability between the diploids, as well as the inflorescence morphology, pollen size, meiotic behavior, and fertility of the synthetic polyploids. Morphologically, the synthetics resemble the natural polyploids with short- and long-liguled forms of T. miscellus resulting when T. pratensis and T. dubius are reciprocally crossed. Synthetic T. mirus was also formed reciprocally, but without any obvious morphological differences resulting from the direction of the cross. Of the 27 original crosses that yielded 171 hybrid individuals, 18 of these lineages have persisted to produce 386 S(1) progeny; each of these lineages has produced S(2) seed that are viable. The successful generation of these synthetic polyploids offers the opportunity for detailed comparative studies of natural and synthetic polyploids within a nonmodel system. PMID- 21628251 TI - A preliminary phylogeny of the 'didymocarpoid Gesneriaceae' based on three molecular data sets: Incongruence with available tribal classifications. AB - The 'didymocarpoid Gesneriaceae' (traditional subfam. Cyrtandroideae excluding Epithemateae) are the largest group of Old World Gesneriaceae, comprising 85 genera and 1800 species. We attempt to resolve their hitherto poorly understood generic relationships using three molecular markers on 145 species, of which 128 belong to didymocarpoid Gesneriaceae. Our analyses demonstrate that consistent topological relationships can be retrieved from data sets with missing data using subsamples and different combinations of gene sequences. We show that all available classifications in Old World Gesneriaceae are artificial and do not reflect natural relationships. At the base of the didymocarpoids are grades of clades comprising isolated genera and small groups from Asia and Europe. These are followed by a clade comprising the African and Madagascan genera. The remaining clades represent the advanced Asiatic and Malesian genera. They include a major group with mostly twisted capsules. The much larger group of remaining genera comprises exclusively genera with straight capsules and the huge genus Cyrtandra with indehiscent fruits. Several genera such as Briggsia, Henckelia, and Chirita are not monophyletic; Chirita is even distributed throughout five clades. This degree of incongruence between molecular phylogenies, traditional classifications, and generic delimitations indicates the problems with classifications based on, sometimes a single, morphological characters. PMID- 21628252 TI - Phylogenetic analysis of sexual systems in Inuleae (Asteraceae). AB - From an ancestor with bisexual flowers, plants with unisexual flowers, or even unisexual individuals have evolved in different lineages of angiosperms. The Asteraceae tribe Inuleae includes hermaphroditic, monoecious, dioecious, and gynomonoecious species. Gynomonoecy, the sexual system in which female and bisexual flowers occur on the same plant, is prevalent in the Asteraceae. We inferred one large gene phylogeny (ndhF) and two supertrees to investigate whether gynomonoecy was a stage in the evolution from hermaphroditism to monoecy. We identified transitions in sexual system evolution using the stochastic character mapping method. From gynomonoecious ancestors, both hermaphroditic and monoecious descendants have evolved. Gynomonoecy was not restricted to a stage in the evolution toward monoecy because the number of transitions and the rate of change from monoecy to gynomonoecy were much higher than the opposite. We also investigated one hypothesized association among female flowers and the development of a petaloid ray as an explanation of gynomonoecy maintenance in Asteraceae. We found that peripheral female flowers and petaloid rays were phylogenetically correlated. However, empirical evidence shows that a causal relationship between these traits is not clear. PMID- 21628253 TI - Molecular phylogenetics of Prescottiinae s.l. and their close allies (Orchidaceae, Cranichideae) inferred from plastid and nuclear ribosomal DNA sequences. AB - The Andes are a cradle of orchid evolution, but most phylogenetic studies of Orchidaceae in this biodiversity hotspot have dealt with epiphytic epidendroid lineages. Here we present a study on neotropical, terrestrial, orchidoid taxa of Prescottiinae s.l. (8 genera, ~100 species), which are adapted to some of the highest elevation habitats on earth that support orchids. They are currently included within an expanded concept of Cranichidinae in the tribe Cranichideae, but DNA sequence data show that neither Prescottiinae s.l. nor Cranichidinae s.s. are monophyletic. Prescottiinae s.l. consist of two strongly supported lineages: the Altensteinia and Prescottia clades, which have closer affinities to Spiranthinae than to Cranichidinae. The Prescottia clade comprises two well supported subclades, one including most sampled species of Prescottia and a second one with Pseudocranichis thysanochila sister to Prescottia tubulosa. As a group, they are sister to Spiranthinae. Sister to this pair is the Altensteinia clade comprised of six genera, whose intergeneric relationships are well resolved. Finally, Cranichidinae s.s. is sister to all three of these clades. Morphological and ecological features distinguishing the major groups are discussed, as are potential synapomorphies to define them. The reconstructed phylogeny indicates that the classification of Cranichideae needs to be reexamined. PMID- 21628254 TI - Global chloroplast phylogeny and biogeography of bracken (Pteridium; Dennstaedtiaceae). AB - Bracken ferns (genus Pteridium) represent an ancient species complex with a natural worldwide distribution. Pteridium has historically been treated as comprising a single species, but recent treatments have recognized several related species. Phenotypic plasticity, geographically structured morphological variation, and geographically biased sampling have all contributed to taxonomic confusion in the genus. We sampled bracken specimens worldwide and used variable regions of the chloroplast genome to investigate phylogeography and reticulate evolution within the genus. Our results distinguish two major clades within Pteridium, a primarily northern hemisphere Laurasian/African clade, which includes all taxa currently assigned to P. aquilinum, and a primarily southern hemisphere Austral/South American clade, which includes P. esculentum and P. arachnoideum. All European accessions of P. aquilinum subsp. aquilinum appear in a monophyletic group and are nested within a clade containing the African P. aquilinum taxa (P. aquilinum subsp. capense and P. aquilinum subsp. centrali africanum). Our results allow us to hypothesize the maternal progenitors of two allotetraploid bracken species, P. caudatum and P. semihastatum. We also discuss the biogeography of bracken in the context of the chloroplast phylogeny. Our study is one of the first to take a worldwide perspective in addressing variation in a broadly distributed species complex. PMID- 21628255 TI - Cone and seed trait variation in whitebark pine (Pinus albicaulis; Pinaceae) and the potential for phenotypic selection. AB - Phenotypic variation among individuals is necessary for natural selection to operate and is therefore essential for adaptive evolution. However, extensive variation within individuals can mask variation among individuals and weaken the potential for selection. Here we quantify variation among and within individuals in female cone and seed traits of whitebark pine (Pinus albicaulis). In many plants, the production of numerous reproductive structures creates the potential for considerable variation within a plant, but these same traits should also undergo strong selection because of their direct link to plant fitness. We found about twice as much variation among individuals (overall mean = 65.3 +/- 4.5% SE) than within individuals (overall mean = 34.7 +/- 4.5%). One only needs to sample three to five cones per tree to accurately assess variation among trees in most cone and seed traits. The ease at which trees can be assessed helps account for the strong and consistent patterns of phenotypic selection exerted by seed predators and dispersers of whitebark pine and many other conifers. In contrast, the few traits where variation within trees equaled or exceeded that among trees underwent weak if any phenotypic selection. PMID- 21628256 TI - Development and morphology of flowers and inflorescences in Balanophora papuana and B. elongata (Balanophoraceae). AB - Extreme modification and reduction in floral morphology presents an obstacle to determining the evolutionary relationships and homologies of the holoparasites in Balanophoraceae. Developing flowers and inflorescences of two dioecious species, Balanophora papuana and B. elongata, were compared to each other and to the monoecious B. fungosa. Intermingled with flowers in the male inflorescences are bracts (B. elongata) or bract parts (B. papuana). In the latter, early cessation of bract tip growth results in two half-bracts, which become displaced during inflorescence elongation, thus disproving the view that these bract-like structures are axial in nature. Male flower primordia emerge in positions axillary to the dividing bracts, and both arise in a spiral sequence. This pattern is modified in B. papuana by the formation of pseudowhorls of four. In both species, the staminate flowers consist of a generally four-merous perianth and a synandrium of congenitally fused stamens. Male flower and bract ontogeny (but not pollen sacs) conform to patterns seen in other angiosperms. More problematic are the carpellate flowers whose primordia arise in irregular order between club-shaped, radially symmetrical organs called claviform bodies. The interpretation that these bodies are homologous to the peltate bracts of Helosideae appears plausible, but cannot explain their nonspiral initiation and radial symmetry. PMID- 21628257 TI - Constraints on leaf structural traits in wetland plants. AB - A plant species' ecology is associated with leaf economics, characterized, e.g., by photosynthetic rate, construction costs, and leaf life span. Specific leaf area (SLA, leaf area per leaf dry mass) is often considered to be a key trait in this respect, explaining interspecific variation in leaf economics. To understand factors constraining the specific leaf area, we investigated size-related biomechanical constraints of the traits that determine the SLA-leaf thickness, leaf dry matter content and leaf density-and the constraints these traits exert on each other among 33 herbaceous wetland species of northern Ontario with a wide variety of leaf forms, ranging from wide laminar leaves to long, narrow, and relatively thick columnar leaves. Data from garden experiments were compared with field data. The results agree with biomechanical predictions that lamina thickness and leaf dry matter content are positively and leaf density (fresh mass per volume) negatively associated with leaf length. The traits also constrain each other, but these intertrait relationships are confounded by interspecific variation in leaf length. We conclude that for a full understanding of the adaptive significance of leaf structural design, it is essential to include leaf size in the considerations. PMID- 21628258 TI - Selenium protects the hyperaccumulator Stanleya pinnata against black-tailed prairie dog herbivory in native seleniferous habitats. AB - Elemental hyperaccumulation in plants is hypothesized to represent a plant defense mechanism. The objective of this study was to determine whether selenium (Se) hyperaccumulation offers plants long-term protection from the black-tailed prairie dog (Cynomys ludovicianus). Prairie dogs are a keystone species. The hyperaccumulator Stanleya pinnata (prince's plume) co-occurs with prairie dogs in seleniferous areas in the western United States. Stanleya pinnata plants pretreated with high or low Se concentrations were planted on two prairie dog towns with different levels of herbivory pressure, and herbivory of these plants was monitored over 2 years. Throughout this study, plants with elevated Se levels suffered less herbivory and survived better than plants with low leaf Se concentrations. This study indicates that the Se in hyperaccumulator S. pinnata protects the plant in its natural habitat from herbivory by the black-tailed prairie dog. The results from this study support the hypothesis that herbivory by prairie dogs or similar small mammals has been a contributing selection pressure for the evolution of plant Se hyperaccumulation in North America. This study is the first to test the ecological significance of hyperaccumulation over a long period in a hyperaccumulator's natural habitat. PMID- 21628259 TI - Temperature requirements differ for the two stages of seed dormancy break in Aegopodium podagraria (Apiaceae), a species with deep complex morphophysiological dormancy. AB - Only a few studies have considered the possibility that low temperature requirements may vary among stages of dormancy break in seeds with morphophysiological dormancy (MPD). We show that this lack of consideration in previous studies on seed dormancy and germination of Aegopodium podagraria might explain the low germination percentages and/or the relatively long periods of incubation needed for germination. Under natural temperatures, embryos began to grow in September and were fully elongated by late December; most growth occurred when the average daily mean temperature was about 10 degrees C. Radicles emerged under snow in late winter, and cotyledons emerged after snowmelt in early spring. In laboratory experiments, 100% of the embryos grew to full length at both 0 and 5 degrees C, whereas 0 degrees C was much more effective than 5 degrees C in overcoming the physiological dormancy in seeds after embryos were fully elongated. Following radicle emergence, cotyledons emerged readily in a wide range of temperatures >=5 degrees C. GA(3) did not substitute for the low temperature requirement for dormancy break. Seed dormancy in A. podagraria fits Nikolaeva's formula for deep complex MPD, i.e., C(3)B-C(3). Better germination of seeds pretreated at 0 degrees than at 5 degrees C has practical implications for cultivating this species. PMID- 21628260 TI - Pollination ecology of the high latitude, dioecious cloudberry (Rubus chamaemorus; Rosaceae). AB - In a 3-yr study, we examined the pollinator guild and intersexual floral characteristics of the dioecious, perennial cloudberry (Rubus chamaemorus), which flowers in early spring. The findings contribute to our general understanding of pollination ecology at high latitudes and provide important information for the commercialization of cloudberry. Female flowers were smaller than males but provided more nectar, although this resource was low in both sexes. Insects from 43 families visited cloudberry flowers, yet four families (Apidae, Halictidae, Muscidae, Syrphidae) represented ca. 87% of all visitors observed. Introduction experiments revealed that apids and muscids are significantly poorer pollinators (based on fruit production) than halictids and syrphids, but when fruit mass or seed set was considered, there were no significant differences between families. Pollinator importance, a product of flower visitation frequency and seed set effectiveness, revealed that the dipterans were of paramount importance to the pollination of cloudberry. Furthermore, they are limited to cloudberry because their lapping mouthparts exclude them from accessing the nutritional rewards of competing Ericaceae flowers. While the total number of pollinator families observed suggest a generalist pollination system, if one considers the dominant pollinators (flies) as a functional group, then this insect-flower relationship could be considered a specialized one. PMID- 21628261 TI - A climatic and taxonomic comparison between leaf litter and standing vegetation from a Florida swamp woodland. AB - One method to determine past climate has been the use of leaf morphological characteristics of fossil leaves quantified using modern climate and canopy leaf characteristics. Fossil assemblages are composed of abscised leaves, and climate may be more accurately determined by using leaves from leaf litter instead of the canopy. To better understand whether taphonomic processes make a difference in this relationship, a north-central Florida woodland was sampled to determine the morphologically based climate estimates from these leaves. Leaves from woody, dicotyledonous plants were collected and identified, then compared using presence/absence data and analyzed using several linear regression equations and the CLAMP data set. Although the majority of standing vegetation was reflected in leaf litter, some inconsistencies were observed, which may reflect plant community structure or sampling technique. Mean annual temperature (MAT) and growing season precipitation (GSP) were estimated from leaf litter morphological characters and living leaves. Overall, values for MAT estimated from litter and living leaves were cooler than actual MATs, although several accurate and high estimates were obtained depending on the predictive method used. Estimated GSP values were higher than actual GSPs. Statistically, no difference was observed between MAT and GSP estimates derived from leaf litter vs. estimates derived from living leaves, with one exception. PMID- 21628262 TI - Patterns of hybridization and introgression between invasive Ulmus pumila (Ulmaceae) and native U. rubra. AB - Natural hybridization between introduced species and their native congeners occurs frequently and can create serious conservation concerns. Ulmus pumila (Siberian elm) is an introduced Asian elm species that has naturalized in the United States and is now considered invasive in 41 states. Red elm (U. rubra), a native to the eastern United States, often occurs in sympatry with Siberian elm, and the two species are thought to hybridize. Here, we genetically characterized reference populations of the two elm species to identify species-specific microsatellite alleles. These markers were used to classify individuals in putative hybrid zones as parental species or hybrids, assess the extent of hybridization, and track patterns of introgression. We identified nine U. rubra, 32 U. pumila, and 51 hybrid individuals in our hybrid zones. Of the 51 hybrids, 35 were classified as first-generation hybrids and 16 as backcrosses. The majority of the backcrosses (88%) were introgressed toward U. pumila. Our classification of genotypes was consistent whether we used manual classification, principal coordinate analyses or Bayesian clustering. We observed greater genetic diversity and new combination of alleles in the hybrids. Our study indicates widespread hybridization between U. pumila and U. rubra and an asymmetric pattern of introgression toward U. pumila. PMID- 21628263 TI - Local adaptation in European populations of Arabidopsis lyrata (Brassicaceae). AB - We studied local adaptation to contrasting environments using an organism that is emerging as a model for evolutionary plant biology-the outcrossing, perennial herb Arabidopsis lyrata subsp. petraea (Brassicaceae). With reciprocal transplant experiments, we found variation in cumulative fitness, indicating adaptive differentiation among populations. Nonlocal populations did not have significantly higher fitness than the local population. Experimental sites were located in Norway (alpine), Sweden (coastal), and Germany (continental). At all sites after one year, the local population had higher cumulative fitness, as quantified by survival combined with rosette area, than at least one of the nonlocal populations. At the Norwegian site, measurements were done for two additional years, and fitness differences persisted. The fitness components that contributed most to differences in cumulative fitness varied among sites. Relatively small rosette area combined with a large number of inflorescences produced by German plants may reflect differentiation in life history. The results of the current study demonstrate adaptive population differentiation in A. lyrata along a climatic gradient in Europe. The studied populations harbor considerable variation in several characters contributing to adaptive population differentiation. The wealth of genetic information available makes A. lyrata a highly attractive system also for examining the functional and genetic basis of local adaptation in plants. PMID- 21628264 TI - Fine-scale genetic structure enhances biparental inbreeding by promoting mating events between more related individuals in wild soybean (Glycine soja; Fabaceae) populations. AB - Outcrossing between genetically distant individuals in a plant population enhances allelic heterozygosity-an important source for genetic diversity and adaptive evolution. Fine-scale spatial genetic structure (FSGS) can interfere with outcrossing by promoting mating between more related individuals. To test the influence of FSGS on outcrossing, FSGS and outcrossing rates were analyzed with four wild soybean (Glycine soja) populations from different habitats, using simple sequence repeat (SSR) fingerprints. Spatial autocorrelation analysis indicated variable FSGS (15.44-25.87 m) in all four populations. Multilocus mixed mating analysis of 1605 progeny indicated substantial variation in single-locus outcrossing (T(s) = 6.3-12.6%) although the total outcrossing rates as estimated by multilocus outcrossing (T(m) = 12.8-17%) did not vary significantly among populations. The comparison between FSGS and outcrossing rates demonstrated that strong FSGS with large genetic patch size can enhance biparental inbreeding by promoting mating between more related individuals in a population. The results suggest that patch size management can aid in situ conservation by avoiding formation of strong FSGS and encouraging true outcrossing among individuals. PMID- 21628265 TI - Reproductive implications of combined and separate sexes in a trioecious population of Opuntia robusta (Cactaceae). AB - Opuntia robusta has hermaphroditic, dioecious, and trioecious populations. To enhance our understanding of this breeding system diversity, we compared the reproductive output of males, females, and hermaphrodites in a trioecious population using field evaluations, controlled crosses, and progeny tests. Unisexuals were fully sterile in one sex function. Hermaphrodites were fully fertile for both functions. Consistent with the sex-allocation theory, unisexuality increased the quality and quantity (in males) of the gametes of the functional sex, relative to those of hermaphrodites, probably explained by maternal and paternal effects. The increase was higher in males than in females, suggesting a more expensive female function. Theoretically, this disproportional increase is required for unisexuals to invade a hermaphroditic population with prior selfing, negligible pollen discounting, and undetectable inbreeding depression, features found in O. robusta, therefore helping to explain dioecious populations. However, in the study population, the actual seed output of females was lower and had a higher variance than that of hermaphrodites, which also reproduce through pollen. Unisexuals are unlikely to be maintained by their actual reproductive output in this pollen-limited environment. Hermaphrodites may persist in this population by producing their seeds autonomously and by reducing interspecific fertilization by prior selfing and ovule discounting. PMID- 21628266 TI - Ovule number per flower in a world of unpredictable pollination. AB - The number of ovules per flower varies over several orders of magnitude among angiosperms. Here we consider evidence that stochastic uncertainty in pollen receipt and ovule fertilization has been a selective factor in the evolution of ovule number per flower. We hypothesize that stochastic variation in floral mating success creates an advantage to producing many ovules per flower because a plant will often gain more fitness from occasional abundant seed production in randomly successful flowers than it loses in resource commitment to less successful flowers. Greater statistical dispersion in pollination and fertilization among flowers increases the frequency of windfall success, which should increase the strength of selection for greater ovule number per flower. We therefore looked for evidence of a positive relationship between ovule number per flower and the statistical dispersion of pollen receipt or seed number per flower in a comparative analysis involving 187 angiosperm species. We found strong evidence of such a relationship. Our results support the hypothesis that unpredictable variation in mating success at the floral level has been a factor in the evolution of ovule packaging in angiosperms. PMID- 21628267 TI - Evolution of autonomous selfing accompanies increased specialization in the pollination system of Schizanthus (Solanaceae). AB - The co-occurrence of elaborate flowers visited by specific groups of pollinators and capacity for autonomous selfing in the same plant species has puzzled evolutionary biologists since the time of Charles Darwin. To examine whether autonomous selfing and floral specialization evolved in association, we quantified the autofertility level (AFI) in nine Schizanthus species characterized by a wide range of pollination specialization, revealing AFI values of 0.02 to complete selfing. An independent contrasts analysis conducted on AFIs and number of functional pollinator groups showed that autonomous selfing evolved from an ancestral outcrossing system as plants became increasingly specialized (r = -0.82). To assess whether autonomous selfing together with specialization acts as a reproductive assurance mechanism, we estimated spatial and interannual variation in fruit set due to pollinator failure in two closely related high Andean Schizanthus species differing in their specialization levels. Variation in pollinator failure rate was more pronounced and autonomous selfing increased fruit production over biotically assisted pollination in the more specialized species. Our study suggests that specialized pollination deems species more vulnerable to pollinator fluctuation thus promoting the evolution of delayed autonomous selfing. PMID- 21628268 TI - DNA barcoding will frequently fail in complicated groups: An example in wild potatoes. AB - DNA barcoding ("barcoding") has been proposed as a rapid and practical molecular method to identify species via diagnostic variation in short orthologous DNA sequences from one or a few universal genomic regions. It seeks to address in a rapid and simple way the "taxonomic impediment" of a greater need for taxonomic identifications than can be supplied by taxonomists. Using a complicated plant group, Solanum sect. Petota (wild potatoes), I tested barcoding with the most variable and frequently suggested plant barcoding regions: the internal nontranscribed spacer of nuclear ribosomal DNA (ITS) and the plastid markers trnH psbA intergenic spacer and matK. These DNA regions fail to provide species specific markers in sect. Petota because the ITS has too much intraspecific variation and the plastid markers lack sufficient polymorphism. The complications seen in wild potatoes are common in many plant groups, but they have not been assessed with barcoding. Barcoding is a retroactive procedure that relies on well defined species to function, is based solely on a limited number of DNA sequences that are often inappropriate at the species level, has been poorly tested with geographically well-dispersed replicate samples from difficult taxonomic groups, and discounts substantial practical and theoretical problems in defining species. PMID- 21628269 TI - Genetic diversity and structure in semiwild and domesticated chiles (Capsicum annuum; Solanaceae) from Mexico. AB - The chile of Mesoamerica, Capsicum annuum, is one of five domesticated chiles in the Americas. Among the chiles, it varies the most in size, form, and color of its fruits. Together with maize, C. annuum is one of the principal elements of the neotropical diets of Mesoamerican civilizations. Despite the great economic and cultural importance of C. annuum both worldwide and in Mexico, however, very little is known about its geographic origin and number of domestications. Here we sampled a total of 80 accessions from Mexico (58 semiwild and 22 domesticated) and examined nucleotide sequence diversity at three single- or low-copy nuclear loci, Dhn, G3pdh, and Waxy. Across the three loci, we found an average reduction of ca. 10% in the diversity of domesticates relative to semiwild chiles and geographic structure within Mexican populations. The Yucatan Peninsula contained a large number of haplotypes, many of which were unique, suggesting an important region of chile domestication and center of diversity. The present sampling of loci did not conclusively resolve the number and location of domestications, but several lines of evidence suggest multiple independent domestications from widely distributed progenitor populations. PMID- 21628270 TI - Moving with climbing plants from Charles Darwin's time into the 21st century. AB - We provide an overview of research on climbing plants from Charles Darwin to the present day. Following Darwin's interests, this review will focus on functional perspectives including attachment mechanisms and stem structure and function. We draw attention to a number of unsolved problems inviting future research. These include the mechanism for establishment of the twining habit, a quantitative description following the development of a tissue element through space and time, the chemistry of sticky exudates, the microstructure of xylem and the capacity for water storage, the vulnerability to embolism, and the mechanism for embolism repair. In conclusion we cite evidence that, in response to increasing CO(2) concentration, anthropic perturbation and/ or increasing forest fragmentation, lianas are increasing relative to tree species. In the 21st century, we are returning to the multiscale, multidisciplinary approach taken by Darwin to understand natural history. PMID- 21628271 TI - Grasses of different C4 subtypes reveal leaf traits related to drought tolerance in their natural habitats: Changes in structure, water potential, and amino acid content. AB - Three grasses (Poaceae) of different C(4) subtypes, Paspalum dilatatum (NADP malic enzyme [ME]), Cynodon dactylon (NAD-ME) and Zoysia japonica (phosphoenolpyruvate carboxykinase), occur in natural habitats that differ in annual rainfall. Their leaf characteristics were studied to identify traits related to drought tolerance. Plants were grown in pots, and water deficit was gradually induced by withholding water. Leaves of Z. japonica had the greatest and P. dilatatum the lowest relative dry matter content. Transverse sections of leaves that developed during the water deficit showed little change compared to control leaves, consistent with low phenotypic plasticity. Anatomical features distinguished the three species, with xeromorphic characteristics most strongly represented in Z. japonica. The leaf relative water content (RWC) decreased with the soil water content similarly for the three grasses. However, at 80% RWC, the leaf water potential was -3.1 MPa for Z. japonica and only -1.3 MPa for P. dilatatum and C. dactylon. Soluble amino acids, especially proline, increased as RWC decreased in leaves of C. dactylon and Z. japonica. Phenylalanine, valine, leucine, and isoleucine increased more in Z. japonica than in the other two species. The results provide evidence that C. dactylon and, especially, Z. japonica have evolved leaf traits better suited to arid habitats. PMID- 21628272 TI - Floral organogenesis and the developmental basis for pollinator deception in the asiatic dayflower, Commelina communis (Commelinaceae). AB - The upper half of flowers in Commelina communis deceptively lures potential pollinators with its showy petals and staminodes on the false promise of abundant pollen. This paper presents evidence that staminodization in the upper half is associated with a severe retardation of the entire upper floral hemisphere early in development. Possible consequences of this developmental retardation are seen also in the gynoecium, where the upper carpel of the three-carpellate ovary is underdeveloped and sterile at maturity. Only late in development do the upper petals and staminodes expand and acquire pigments necessary for their attractive function. We surmise that retardations of this severity are unlikely to be found for functionally fertile organs such as stamens and ovule-producing carpels, because key preparatory events preceding sporogenesis might otherwise be disrupted. Such differential growth about the floral apex resembles that known in some eudicots to be regulated by the TCP gene family; thus, future molecular developmental studies in Commelina may help to extend our understanding of the evolutionary genetics of floral monosymmetry to monocots. PMID- 21628273 TI - Diversity and distribution of idioblasts producing calcium oxalate crystals in Dieffenbachia seguine (Araceae). AB - Although cells that synthesize crystals are known throughout the plant kingdom, their functional significance is still unknown. Mechanical support, mineral balance, waste sequestration, and protection against herbivores have all been proposed as crystal functions. To seek clues to their role(s), I systematically examined all organs except fruit of Dieffenbachia seguine (Araceae) for crystals. Crystals were found in nearly every organ. Raphides (long, slim, pointed crystals) were most common, but druses (crystal aggregates) and prisms were also found. Raphides varied in size by a factor of 10 and also in organization from tightly bundled to loosely organized. Biforines, a type of cell capable of expelling raphides, or biforine-like cells, were found in nearly all organs, but especially in leaves, spathes, and anthers. Different organs had different crystal complements, and characteristic crystals were found at specific locations, such as among pollen, along the undersides of leaf veins, and at root branch points. All crystals appeared to be composed of calcium oxalate, based on acid solubility. Possible roles of the crystals are discussed in light of these findings. PMID- 21628274 TI - Seed tolerance to predation: Evidence from the toxic seeds of the buckeye tree (Aesculus californica; Sapindaceae). AB - Tolerance, the capacity of plants to withstand attack by animals, as opposed to resistance, has been poorly examined in the context of seed predation. We investigated the role that the seed mass of the large-seeded endemic tree Aesculus californica plays as a tolerance trait to rodent attack by comparing, under greenhouse conditions, patterns of germination, and subsequent seedling growth, of seeds with a wide range of natural damage. Germination percentage was reduced by 50% and time to germination by 64% in attacked compared to intact seeds, and germination probability was negatively correlated with damage. Seedlings that emerged from intact seeds were taller and bore more leaves than those from damaged seeds. This species' large seed mass favors tolerance to damage because heavily damaged seeds are able to germinate and produce seedlings. This finding is significant given that seeds of this species are known to contain chemical compounds toxic to vertebrates, a resistance trait. We posit that this combination of tolerance and resistance traits might be a particularly effective antipredation strategy when seeds are exposed to a variety of vertebrate predators. PMID- 21628275 TI - Multiple origins promote the ecological amplitude of allopolyploid Aegilops (Poaceae). AB - Polyploidy has been ubiquitous in plant evolution and is thought to be an important engine of biodiversity that facilitates speciation, adaptation, and range expansion. Polyploid species can exhibit higher ecological tolerance than their progenitor species. For allotetraploid species, this higher tolerance is often attributed to the existence of heterosis resulting from entire genome duplication. However, multiple origins of allopolyploid species may further promote their ecological success by providing genetic variability in ecological traits underlying local adaptation and range expansion. Here we show in a group of allopolyploid species in the genus Aegilops that range size and abundance are correlated with the number of inferred origins. We found that allopolyploid Aegilops spp. contain multiple chloroplast haplotypes, each identical to haplotypes of the diploid progenitor species, indicating multiple origins as the major source of variation. The number of inferred origins in each allopolyploid species was correlated to the total area occupied by the allopolyploid and the tendency for the species to be common. Additionally, we found differences in ecological tolerance among independent origins in Aegilops triuncialis. These results strongly support the hypothesis that the introduction of genetic variability by multiple origins can increase the ecological amplitude and evolutionary success of allopolyploid species. PMID- 21628276 TI - Pathogenesis of bryophyte hosts by the ascomycete Atradidymella muscivora. AB - Atradidymella muscivora (Pleosporales) is a bryophyte pathogen that infects the mosses Aulacomnium palustre, Hylocomium splendens, and Polytrichum juniperinum. Light and scanning electron microscopy and extracellular enzyme production were used to characterize the interactions between this fungus and its native hosts and the model host Funaria hygrometrica. Penetration was direct via hyphae or appressoria, and hosts responded by forming layered, darkly pigmented deposits at penetration sites, similar to the papillae formed by vascular plants in response to fungal infection. Infected hosts gradually became chlorotic as hyphae grew intracellularly, presumably killing host cells. Pycnidia of the Phoma anamorph (P. muscivora) and uniloculate pseudothecia were initiated as tightly packed masses of stromatic dematiaceous hyphae within a single host cell. Mature pycnidia and pseudothecia were erumpent. A new microniche among bryophilous fungi is described, whereby A. muscivora supplants the gemmae of Aul. palustre and exploits the normal nutrient-flow of the moss gametophyte. Atradidymella muscivora produced both cellulases and soluble polyphenolic oxidases, allowing it to also function as a saprobe and degrade the cell walls of bryophytes. The saprophytic and pathogenic abilities of A. muscivora suggest it may play a role in nutrient cycling, population dynamics, and small-scale disturbances in boreal ecosystems. PMID- 21628277 TI - Atradidymella muscivora gen. et sp. nov. (Pleosporales) and its anamorph Phoma muscivora sp. nov.: A new pleomorphic pathogen of boreal bryophytes. AB - During a survey of bryophilous fungi from boreal and montane habitats, 12 isolates of a hitherto unknown plant pathogenic member of the Pleosporales were recovered from Aulacomnium palustre, Hylocomium splendens, and Polytrichum juniperinum, and described as Atradidymella muscivora gen. et sp. nov. Atradidymella is characterized by minute, unilocular, setose pseudothecia having 2-3 wall layers; brown, fusiform, 1-septate ascospores; and a Phoma anamorph. The genus is distinguished from all other pleosporalean genera with brown, fusiform ascospores on the basis of ascospore and pseudothecium morphology and a highly reduced stroma that is localized within a single host cell. Atradidymella muscivora is distinguished by its minute pseudothecia (<115 MUm) and ascospores that are slightly allantoid and constricted at the septum with the upper cell often wider than the lower. Its anamorph, Phoma muscivora sp. nov., is morphologically distinguishable from P. herbarum in having smaller conidia. Parsimony analysis of the ITS rDNA region indicates A. muscivora has affinities to the Phoma-Ascochyta-Didymella clade that is sister to the Phaeosphaeriaceae within the Pleosporales. PMID- 21628278 TI - Phylogenetic diversification of Equisetum (Equisetales) as inferred from Lower Cretaceous species of British Columbia, Canada. AB - Three types of anatomically preserved vegetative shoots with features that characterize crown group Equisetum have been discovered in Lower Cretaceous deposits (~136 Ma) of British Columbia, Canada, suggesting the genus is much older than currently believed. Specimens include two types of aerial shoots described as E. haukeanum sp. nov. and E. vancouverense sp. nov. and one type of subterranean rhizome. Shoots are 1-2 mm in diameter, jointed, and in cross section have fluted stems with a hollow pith. Distinctive patterns of cortical sclerenchyma and different ridge morphologies characterize each shoot morphotype. Nodes display irregular branching, highly fused leaf sheaths, and a nodal diaphragm. The aerial stem morphospecies have vallecular canals on alternating radii with carinal canals of an equisetostele surrounded by only a few tracheids. No secondary tissues are produced. Bands of surficial stomata flank the furrows of one morphospecies. Rhizomes and aerial shoots are of a similar size, suggesting that the plants were equivalent in stature to the smallest living Equisetum species. These fossils augment our understanding of evolutionary transformations that led from Paleozoic Archaeocalamitaceae and Calamitaceae to crown group Equisetaceae, suggesting that the initial diversification of Equisetum began far earlier than suggested by molecular-clock-based estimates. PMID- 21628279 TI - Palms (Arecaceae) from a Paleocene rainforest of northern Colombia. AB - Palms are a monophyletic group with a dominantly tropical distribution; however, their fossil record in low latitudes is strikingly scarce. In this paper, we describe fossil leaves, inflorescences, and fruits of palms from the middle to late Paleocene Cerrejon Formation, outcropping in the Rancheria River Valley, northern Colombia. The fossils demonstrate the presence of at least five palm morphospecies in the basin ca. 60 Ma. We compare the morphology of the fossils with extant palms and conclude that they belong to at least three palm lineages: the pantropical Cocoseae of the subfamily Arecoideae, the monotypic genus Nypa, and either Calamoideae or Coryphoideae. The fossil fruits and inflorescences are among the oldest megafossil records of these groups and demonstrate that the divergence of the Cocoseae was more than 60 Ma, earlier than has previously been thought. These fossils are useful in tracing the range expansion or contraction of historical or current neotropical elements and also have profound implications for the understanding of the evolution of neotropical rainforests. PMID- 21628280 TI - A new method to measure leaf age: Leaf measuring-interval index. AB - We propose a new method, the leaf measuring-interval index (LMI), to estimate leaf age in morphological and physiological studies of leaves. When the plastochron, the interval between the initiation of successive leaves, is constant, the well-known leaf plastochron index (LPI) provides a robust measure of leaf age. When the duration of the plastochron is not uniform, however, we show that the LPI can (in simulations) and does (with actual data) turn variation in duration of the plastochron into variance about the regression estimates of leaf growth curves. The method we present in this paper, the LMI, is plastochron independent. This new method is particularly suited, therefore, for studies of plants growing in natural environments rather than in controlled growth facilities where the assumptions of the LPI method can be met. PMID- 21628281 TI - Phytochrome gene expression and phylogenetic analysis in the short-day plant Pharbitis nil (Convolvulaceae): Differential regulation by light and an endogenous clock. AB - To investigate the role of distinct phytochrome pools in photoperiodic timekeeping, we characterized four phytochrome genes in the short-day plant Pharbitis nil. Each PHY gene had different photosensory properties and sensitivity to night break that inhibits flowering. During extended dark periods, PHYE, PHYB, and PHYC mRNA accumulation exhibited a circadian rhythmicity indicative of control by an endogenous clock. Phylogenetic analysis recovered four clades of angiosperm phytochrome genes, phyA, phyB, phyC, and phyE. All except the phyE clade included sequences from both monocots and eudicots. In addition, phyA is sister to phyC and phyE sister to phyB, with gymnosperm sequences sister to either the phyA-phyC clade or to the phyB-phyE clade. These results suggest that a single duplication occurred in an ancestral seed plant before the divergence of extant gymnosperms from angiosperms and that two subsequent duplications occurred in an ancestral angiosperm before the divergence of monocots from eudicots. Thus in P. nil, a multigene family with different patterns of mRNA abundance in light and darkness contributes to the total phytochrome pool: one pool is light labile (phyA), whereas the other is light stable (phyB and phyE). In addition, PHYC mRNA represents a third phytochrome pool with intermediate photosensory properties. PMID- 21628282 TI - Comparative analysis of the reproductive ecology of Monotropa and Monotropsis: Two mycoheterotrophic genera in the Monotropoideae (Ericaceae). AB - Studies of mycoheterotrophs, defined as plants that obtain carbon resources from associated mycorrhizal fungi, have fundamentally contributed to our understanding of the importance and complexity of symbiotic ecological interactions. However, to date, the reproductive ecology of these organisms remains empirically understudied, with existing literature presenting hypotheses about traits including a generalist pollination syndrome and autogamous self-pollination. To address this gap in our knowledge of the reproductive ecology of mycoheterotrophic plants, we comparatively analyzed three species of two monotropoid genera, Monotropa and Monotropsis. During three consecutive years of field observations and manipulations of four populations of Monotropa uniflora, seven of M. hypopitys (both red and yellow color forms), and two of Monotropsis odorata, we investigated flowering phenology, pollination ecology, breeding system, floral herbivory, and reproductive effort and output. Contrary to previous predictions, our results revealed that taxa are largely outcross pollinated and specialized toward Bombus pollinators. Additionally, species differ in breeding system, timing and duration of reproductive development, fluctuations in reproductive effort and output, and fitness impacts of herbivory. This study is the first thorough investigation of the reproductive ecology of mycoheterotrophic species and provides insight into possible limitations in reproductive traits imposed by a mycoheterotrophic life history. PMID- 21628283 TI - Molecular taxonomy of Camellia (Theaceae) inferred from nrITS sequences. AB - Camellia, comprising more than 200 species, is the type genus of the family Theaceae. Currently, the interspecies relationship of the economically important genus is still a matter of great debate and controversy. In an attempt to help settle this dispute using molecular phylogeny, we analyzed ITS sequences of 112 species of Camellia. The maximum parsimony and Bayesian trees grouped these species into eight major clades and four isolates. The current study supported the monophyly of sections Thea and Furfuracea, a merged section of Theopsis and Eriandra and the formation of section Oleifera by H, -t. Chang (Flora of Reipulicae Popularis Sinicae. Tomus 49 (3), Science Press, China). The study suggested the polyphyletic nature of the sections Camellia, Paracamellia, Pseudocamellia, and Tuberculata and the paraphyletic nature of the section Chrysantha but did not support the sectional status of the three small sections, Archecamellia, Piquetia, and Sterocarpus. We also discuss the results in terms of morphology, geographic distribution and the results from an earlier molecular phylogeny analysis. PMID- 21628284 TI - Perianth evolution in the sandalwood order Santalales. AB - Flowers of Santalales remain largely unexplored with several questions of homology unanswered despite the large size of the order. Morphological and ontogenetic floral studies have the potential to identify new informative characters. We studied floral development in species of Loranthaceae, Santalaceae, Opiliaceae, and "Olacaceae" with scanning electron microscopy to clarify the origin and evolution of the perianth in Santalales. The perianth is either dichlamydeous or when monochlamydeous interpreted as sepals, petals, or tepals. A girdling calyculus of debatable origin is found in some clades. We show that species of Diogoa and Heisteria in "Olacaceae" have a dichlamydeous perianth, unlike Olax in which the calyx is replaced by a calyculus. The calyculus arises by development of two lateral primordia, supporting the hypothesis of bracteole origin. A calyculus with similar development is present in species of Loranthaceae and possibly of Opiliaceae, suggesting a position of Olax closer to these families than to traditional genera of "Olacaceae". The monochlamydeous perianth in Santalaceae is shown to correspond to petals of other members of Santalales. Flower ontogenetic evidence suggests a repeated loss of the calyx, replacement by a calyculus, and further loss, leading to monochlamydeous perianths in Santalaceae. PMID- 21628285 TI - C3 photosynthesis in Aristida longifolia: Implication for photosynthetic diversification in Aristidoideae (Poaceae). AB - Only a small percentage of plant species undergo C(4) photosynthesis. Despite its rarity, the C(4) pathway has evolved numerous times from C(3) ancestors, with as many as 18 independent origins in grasses alone. We report non-Kranz (C(3)) anatomy in Aristida longifolia, a species in a genus of ca. 300 species previously thought to possess only Kranz (C(4)) anatomy. Leaf blade transections of A. longifolia show widely spaced vascular bundles, nonradiate chlorenchyma, and few or no chloroplasts in cells of the sheaths surrounding the vascular bundle, all features indicative of C(3) photosynthesis. Carbon isotope ratios range from -27.68 to -29.71%, likewise indicative of C(3) photosynthesis. We also reconstruct the phylogeny of Aristidoideae, comprising Aristida, Sartidia (C(3)), and Stipagrostis (C(4)), using a sample of 11 species, including A. longifolia, and DNA sequences of the nuclear ribosomal internal transcribed spacer region and the chloroplast rpl16 intron and trnL-trnF region. Sartidia and Stipagrostis resolve as sisters, and sister to this clade is Aristida. Aristida longifolia resolves as sister to the remaining species in the genus. C(3) photosynthesis is hypothesized to be ancestral in Aristidoideae, which means the C(4) pathway evolved twice in the subfamily-in Stipagrostis and early in the diversification of the Aristida clade. PMID- 21628286 TI - Wood anatomy and wood density in shrubs: Responses to varying aridity along transcontinental transects. AB - Wood density plays a key role in ecological strategies and life history variation in woody plants, but little is known about its anatomical basis in shrubs. We quantified the relationships between wood density, anatomy, and climate in 61 shrub species from eight field sites along latitudinal belts between 31 degrees and 35 degrees in North and South America. Measurements included cell dimensions, transverse areas of each xylem cell type and percentage contact between different cell types and vessels. Wood density was more significantly correlated with precipitation and aridity than with temperature. High wood density was achieved through reductions in cell size and increases in the proportion of wall relative to lumen. Wood density was independent of vessel traits, suggesting that this trait does not impose conduction limitations in shrubs. The proportion of fibers in direct contact with vessels decreased with and was independent of wood density, indicating that the number of fiber-vessel contacts does not explain the previously observed correlation between wood density and implosion resistance. Axial and radial parenchyma each had a significant but opposite association with wood density. Fiber size and wall thickness link wood density, life history, and ecological strategies by controlling the proportion of carbon invested per unit stem volume. PMID- 21628287 TI - What is disjunctive xylem parenchyma? A case study of the African tropical hardwood Okoubaka aubrevillei (Santalaceae). AB - The morphological variation and structure-function relationships of xylem parenchyma still remain open to discussion. We analyzed the three-dimensional structure of a poorly known type of xylem parenchyma with disjunctive walls in the tropical hardwood Okoubaka aubrevillei (Santalaceae). Disjunctive cells occurred among the apotracheal parenchyma cells and at connections between axial and ray parenchyma cells. The disjunctive cells were partly detached one from another, but their tubular structures connected them into a continuous network of axial and ray parenchyma. The connecting tubules had thick secondary walls and simple pits with plasmodesmata at the points where one cell contacted a tubule of another cell. The imperforate tracheary elements of the ground tissue were seven times longer than the axial parenchyma strands, a fact that supports a hypothesis that parenchyma cells develop disjunctive walls because they are pulled apart and partly separated during the intrusive growth of fibers. We discuss unresolved details of the formation of disjunctive cell walls and the possible biomechanical advantage of the wood with disjunctive parenchyma: the proportion of tissue that improves mechanical strength is increased by the intrusive elongation of fibers (thick-walled tracheids), whereas the symplastic continuum of the parenchyma is maintained through formation of disjunctive cells. PMID- 21628288 TI - Do lichens domesticate photobionts like farmers domesticate crops? Evidence from a previously unrecognized lineage of filamentous cyanobacteria. AB - Phylogenetic diversity of lichen photobionts is low compared to that of fungal counterparts. Most lichen fungi are thought to be associated with just four photobiont genera, among them the cyanobacteria Nostoc and Scytonema, two of the most important nitrogen fixers in humid ecosystems. Although many Nostoc photobionts have been identified using isolated cultures and sequences, the identity of Scytonema photobionts has never been confirmed by culturing or sequencing. We investigated the phylogenetic placement of presumed Scytonema photobionts and unicellular morphotypes previously assigned to Chroococcus, from tropical Dictyonema, Acantholichen, Coccocarpia, and Stereocaulon lichens. While we confirm that filamentous and unicellular photobiont morphotypes belong to a single clade, this clade does not cluster with Scytonema but represents a novel, previously unrecognized, highly diverse, exclusively lichenized lineage, for which the name Rhizonema is available. The phylogenetic structure observed in this novel lineage suggests absence of coevolution with associated mycobionts at the species or clade level. Instead, highly efficient photobiont strains appear to have evolved through photobiont sharing between unrelated, but ecologically similar, coexisting lineages of lichenized fungi ("lichen guilds"), via the selection of particular photobiont strains through and subsequent horizontal transfer among unrelated mycobionts, a phenomenon not unlike crop domestication. PMID- 21628289 TI - MADS-box gene expression and implications for developmental origins of the grass spikelet. AB - Basic questions regarding the origin and evolution of grass (Poaceae) inflorescence morphology remain unresolved, including the developmental genetic basis for evolution of the highly derived outer spikelet organs. To evaluate homologies between the outer sterile organs of grass spikelets and inflorescence structures of nongrass monocot flowers, we describe expression patterns of APETALA1/FRUITFULL-like (AP1/FUL) and LEAFY HULL STERILE-like (LHS1) MADS-box genes in an early-diverging grass (Streptochaeta angustifolia) and a nongrass outgroup (Joinvillea ascendens). AP1/FUL-like genes are expressed only in floral organs of J. ascendens, supporting the hypothesis that they mark the floral boundary in nongrass monocots, and JaLHS1/OsMADS5 is expressed in the inner and outer tepals, stamen filaments and pistil. In S. angustifolia, SaFUL2 is expressed in all 11 (or 12) bracts of the primary inflorescence branch, but not in the suppressed floral bract below the abscission zone. In contrast, SaLHS1 is only expressed in bracts 6-11 (or 12). Together, these data are consistent with the hypotheses that (1) bracts 1-5 of S. angustifolia primary inflorescence branches and glumes of grass spikelets are homologous and that (2) the outer tepals of immediate grass relatives, bracts 6-8 of S. angustifolia, and the lemma/palea are homologous, although other explanations are possible. PMID- 21628290 TI - Emergent properties of plants competing in silico for space and light: Seeing the tree from the forest. AB - A spatially explicit, reiterative algorithm (SERA) is presented and used to predict multiple aspects of plant population and community dynamics. Using simple physical principles and empirically derived relationships, SERA provides an analytical venue to test alternative hypotheses about individual functional traits governing ecological or evolutionary processes at the population or community level of complexity. Our analyses show that, as a result of competition for light and space, individual-level features scale up to produce species ensemble properties such as the scaling of self-thinning, size-dependent mortality, realistic size-frequency distributions, and a broad spectrum of empirically observed relationships for the species examined. SERA also predicts the competitive exclusion of conifers by angiosperms and the age at which reproductive maturity is achieved by different species. SERA serves as a null hypothesis by demonstrating that biologically complex phenomena, including widely observed scaling relationships at the species-ensemble level, can emerge from the operation of simple and transparent "rules" governing competition for space and light. PMID- 21628291 TI - Water stress tolerance of shrubs in Mediterranean-type climate regions: Convergence of fynbos and succulent karoo communities with California shrub communities. AB - Mediterranean-type climate regions are highly biodiverse and predicted to be particularly sensitive to climate change. Shrubs of the mediterranean-type climate region of South Africa are highly threatened, and their response to water stress has been comparatively little studied. Resistance to water stress induced xylem cavitation (P(50)) and xylem specific hydraulic conductivity (K(s)) were measured in 15 shrub species from fynbos and succulent karoo communities of South Africa. Species displayed a fivefold variation in cavitation resistance (P(50) of -1.9 to -10.3 MPa) with succulent karoo species displaying greater interspecific variability in P(50) than fynbos species. Principal components analysis (including P(50), minimum seasonal water potential, K(s), and xylem density) showed the response to water stress in fynbos species to be similar to chaparral species from the mediterranean-type climate region of California. The data suggest convergence of community and species-specific water stress "strategies" between these mediterranean-type climate regions with respect to their xylem traits. On the basis of the current study and reported plant death and dieback in these regions, woody species within the fynbos may be more susceptible to climate warming and drying than those within the succulent karoo that appear to be utilizing more diverse xylem strategies in response to water stress. PMID- 21628292 TI - Phenotypic plasticity and integration across the canopy of Olea europaea subsp. guanchica (Oleaceae) in populations with different wind exposures. AB - Woody plants, as sessile and long-lived organisms, are expected to have effective mechanisms for dealing with recurrent environmental stresses. In the present study, we hypothesized that phenotypic plasticity (the ability to express alternative phenotypes) and integration (covariation among functionally related traits) are elicited in plants under stressful wind speed conditions. We investigated the within-crown variation of nine vegetative traits of a tree species (Olea europaea subsp. guanchica) in six populations that represented a gradient of wind speed exposures. Wind-exposed twigs in outer-canopy layers had smaller leaves; thinner, lighter, and shorter internodes; and a larger internode cross-sectional area to leaf area ratio. Comparison between field and greenhouse trials revealed that field differences among populations were mediated by phenotypic plasticity. Outer-canopy twigs expressed plastic responses in populations exposed to high wind speeds, whereas inner-canopy twigs displayed high phenotypic convergence among populations. In addition, phenotypic integration increased with wind exposure (outer canopy > inner canopy > greenhouse) and was consequently affected by canopy openness. We conclude that exposure to wind above a certain speed threshold in this woody species elicits a plastic response that is associated with increased integration among traits and involves mechanical and hydraulic rearrangements in more exposed parts of the trees. PMID- 21628293 TI - Evolution of lamina anatomy in the palm family (Arecaceae). AB - The unique properties of tree building in Arecaceae strongly constrain their architectural lability. Potentially compensating for this limitation, the extensive diversification of leaf anatomical structure within palms involves many characters whose alternate states may confer disparate mechanical or physiological capabilities. In the context of a recent global palm phylogeny, we analyzed the evolution of 10 such lamina anatomical characters and leaf morphology of 161 genera, conducting parsimony and maximum likelihood ancestral state reconstructions, as well as tests of correlated evolution. Lamina morphology evolves independently from anatomy. Although many characters do optimize as synapomorphic for major clades, anatomical evolution is highly homoplasious. Nevertheless, it is not random: analyses indicate the recurrent evolution of different cohorts of correlated character states. Notable are two surface layer (epidermis and hypodermis) types: (1) a parallel-laminated type of rectangular epidermal cells with sinuous anticlinal walls, with fibers present in the hypodermis and (2) a cross-laminated type of hexagonal cells in both layers. Correlated with the cross-laminated type is a remarkable decrease in the volume fraction of fibers, accompanied by changes in the architecture and sheath cell type of the transverse veins. We discuss these and other major patterns of anatomical evolution in relation to their biomechanical and ecophysiological significance. PMID- 21628294 TI - Stem architecture in Eleocharis subgenus Limnochloa (Cyperaceae): Evidence of dynamic morphological evolution in a group of pantropical sedges. AB - We examined phylogenetic relationships and patterns of stem structural evolution in Eleocharis subgenus Limnochloa, an ecologically and economically important group of tropical to temperate-growing sedges, whose stems serve as the primary photosynthetic organs. We used maximum parsimony, likelihood, and Bayesian inference to develop phylogenetic trees and stochastic mapping and a Markov one rate model to develop character history reconstructions of stem architecture. A complex history of stem shape evolution characterized by a high degree of homoplasy and rapid rates of change (an average of 13 transitions per character history for about 25 species) was identified across subgenus Limnochloa. Character states transition much more frequently in some lineages than others, but tend to follow a consistent directional pattern of evolutionary change. Our data also suggest that changes in stem shape and anatomy may be associated with speciation events in the subgenus (Pagel's kappa = 0.3503, P = 0.04579) and may have some adaptive significance. The potential adaptive roles of stem structural traits are unclear, but may be elucidated by further studies. This work serves as a starting point for future evolutionary studies of stem shape and structure in monocots and provides important background knowledge for further studies of ecological adaptations of Eleocharis. PMID- 21628295 TI - Molecular and dendrochronological analysis of natural root grafting in Populus tremuloides (Salicaceae). AB - Trembling aspen (Populus tremuloides) is a clonal tree species, which regenerates mostly through root suckering. In spite of vegetative propagation, aspen maintains high levels of clonal diversity. We hypothesized that the maintenance of clonal diversity in this species can be facilitated by integrating different clones through natural root grafts into aspen's communal root system. To verify this hypothesis, we analyzed root systems of three pure aspen stands where clones had been delineated with the help of molecular markers. Grafting between roots was frequent regardless of their genotypes. Root system excavations revealed that many roots were still living below trees that had been dead for several years. Some of these roots had no root connections other than grafts to living ramets of different clones. The uncovered root systems did not include any unique genotypes that would not occur among stems. Nevertheless, acquiring roots of dead trees helps to maintain extensive root systems, which increases the chances of clone survival. Substantial interconnectivity within clones as well as between clones via interclonal grafts results in formation of large genetically diverse physiological units. Such a clonal structure can significantly affect interpretations of diverse ecophysiological processes in forests of trembling aspen. PMID- 21628296 TI - Elucidating the affinities and habitat of ancient, widespread Cyperaceae: Volkeria messelensis gen. et sp. nov., a fossil mapanioid sedge from the Eocene of Europe. AB - The sedges (family Cyperaceae) are an economically and ecologically important monocot group dating back at least to the Paleocene. While modern genera are mostly unknown before the Oligocene, several extinct taxa are recognized as the earliest sedges. Their affinities have been unclear until now, because they are found as isolated, often abraded fruits or endocarps. Exceptionally preserved sedge fossils from the Middle Eocene of Messel, Germany yield more characters for identification. Fossil cyperacean infructescences with in situ pollen are recognized for the first time and show features of the early-divergent mapanioid sedges. Pollen resembles that of tribe Hypolytreae. Comparisons with extant taxa suggest the closest affinities with Hypolytrum and Mapania. However, the Messel fossils represent a distinct taxon, Volkeria messelensis gen. et sp. nov. Without the additional characters of infructescence and pollen, the Messel fruits would have been placed in the extinct genus Caricoidea, a typical Eocene sedge that was widespread across Eurasia. Similarities of fruit structure suggest that Caricoidea was also a mapanioid sedge. Mapanioid sedges are found today in tropical wet forests and swamps, a distribution suggesting that early sedges occupied a similar habitat, unlike many modern sedges, and were not precursors to open grassland vegetation. PMID- 21628297 TI - Tangled trios?: Characterizing a hybrid zone in Castilleja (Orobanchaceae). AB - Hybridization and polyploidization are exceedingly important processes because both influence the ecological envelope and evolutionary trajectory of land plants. These processes are frequently invoked for Castilleja (Indian paintbrushes) as contributors to morphological and genetic novelty and as complicating factors in species delimitations. Here, we provide a detailed analysis of morphological and genetic evidence for hybridization in a well characterized hybrid swarm involving three broadly sympatric species (C. miniata, C. rhexiifolia, C. sulphurea) in western Colorado. Field-classified hybrids are present at high frequencies at these sites and show morphological intermediacy to and segregate for chloroplast DNA haplotypes with C. rhexiifolia and C. sulphurea. Contrarily, DNA content and AFLP variation show that field-classified hybrids are not recent hybrids but a distinctive fourth taxon. Actual hybrids (plants showing admixture >=10% for two genotypic groups) comprised 13% of our sample, with most admixture involving C. rhexiifolia, C. sulphurea, and the unknown taxon. The identity of the field-classified "hybrids" remains unknown; they either represent a stabilized hybrid species or a species with uncharacteristically high diversity for color alleles. This study highlights the importance of examining concordance and discordance between morphology, cytology, and genetic criteria to understand the complex evolutionary history of diverse groups such as Castilleja. PMID- 21628298 TI - Shaping of genetic structure along Pleistocene and modern river systems in the hydrochorous riparian azalea, Rhododendron ripense (Ericaceae). AB - To determine the effects of hydrochory on the formation of the present range of a species and the spatial distribution of genetic variation, we assessed the rangewide genetic structure of a hydrochorous riparian Japanese species (Rhododendron ripense) using four nuclear microsatellite loci. The patterns of isolation by distance and Bayesian clustering analyses of 33 populations suggested that the present range, characterized by both localized and disjunct distributions across the sea, arose from two contrasting colonization events: (1) primary colonization along two Pleistocene rivers that have been submerged and become partly isolated by marine transgression by 6000 years ago, and (2) additional range expansions from these rivers into unconnected neighboring rivers as a result of river captures. Along the Pleistocene rivers, frequent gene flow by hydrochory resulted in the retention of considerable genetic diversity within each population and genetic homogenization among populations. Within unconnected neighboring rivers, genetic diversity was also retained by the simultaneous redistribution of many individuals as a result of river captures, whereas restricted gene flow within a river resulted in genetic divergence among the river populations. Thus, the evolutionary history of hydrochorous R. ripense appears to have been strongly shaped by both ancient and modern rivers. PMID- 21628299 TI - Breeding system and pollination ecology of introduced plants compared to their native relatives. AB - Identifying how plant-enemy interactions contribute to the success of introduced species has been a subject of much research, while the role of plant-pollinator interactions has received less attention. The ability to reproduce in new environments is essential for the successful establishment and spread of introduced species. Introduced plant species that are not capable of autonomous self-fertilization and are unable to attract resident pollinators may suffer from pollen limitation. Our study quantifies the degree of autogamy and pollination ecology of 10 closely related pairs of native and introduced plant species at a single site near St. Louis, Missouri, USA. Most of these species pairs had similar capacities for autogamy; however, of those that differed, the introduced species were more autogamous than their native congeners. Most introduced plants have pollinator visitation rates similar to those of their native congeners. Of the 20 species studied, only three had significant pollen limitation. We suggest that the success of most introduced plant species is because they are highly autogamous or because their pollinator visitation rates are similar to those of their native relatives. Understanding and identifying traits related to pollination success that are key in successful introductions may allow better understanding and prediction of biological invasions. PMID- 21628300 TI - Malpighiales phylogenetics: Gaining ground on one of the most recalcitrant clades in the angiosperm tree of life. AB - The eudicot order Malpighiales contains ~16000 species and is the most poorly resolved large rosid clade. To clarify phylogenetic relationships in the order, we used maximum likelihood, Bayesian, and parsimony analyses of DNA sequence data from 13 gene regions, totaling 15604 bp, and representing all three genomic compartments (i.e., plastid: atpB, matK, ndhF, and rbcL; mitochondrial: ccmB, cob, matR, nad1B-C, nad6, and rps3; and nuclear: 18S rDNA, PHYC, and newly developed low-copy EMB2765). Our sampling of 190 taxa includes representatives from all families of Malpighiales. These data provide greatly increased support for the recent additions of Aneulophus, Bhesa, Centroplacus, Ploiarium, and Rafflesiaceae to Malpighiales; sister relations of Phyllanthaceae + Picrodendraceae, monophyly of Hypericaceae, and polyphyly of Clusiaceae. Oxalidales + Huaceae, followed by Celastrales are successive sisters to Malpighiales. Parasitic Rafflesiaceae, which produce the world's largest flowers, are confirmed as embedded within a paraphyletic Euphorbiaceae. Novel findings show a well-supported placement of Ctenolophonaceae with Erythroxylaceae + Rhizophoraceae, sister-group relationships of Bhesa + Centroplacus, and the exclusion of Medusandra from Malpighiales. New taxonomic circumscriptions include the addition of Bhesa to Centroplacaceae, Medusandra to Peridiscaceae (Saxifragales), Calophyllaceae applied to Clusiaceae subfamily Kielmeyeroideae, Peraceae applied to Euphorbiaceae subfamily Peroideae, and Huaceae included in Oxalidales. PMID- 21628301 TI - Population structure and phylogeography of the mistletoes Tristerix corymbosus and T. aphyllus (Loranthaceae) using chloroplast DNA sequence variation. AB - The mistletoe Tristerix corymbosus (Loranthaceae) is present in the temperate forest and Chilean matorral biomes of Chile and northwest Patagonia. The closely related cactus-specific species, T. aphyllus, occurs only in the matorral biome. The population structure of these mistletoes was examined to determine whether the distribution of haplotypes corresponds mostly to geographic zone, biome, or other biotic factors. Samples from 108 individuals in 26 localities of T. corymbosus and 13 individuals in four localities of T. aphyllus were collected. Sequences were obtained from two chloroplast genome regions: the atpB-rbcL spacer and the trnL-F region. Haplotypes were analyzed using parsimony and Bayesian trees as well as parsimony networks. All methods placed the haplotypes in four clades, one of which corresponded to T. aphyllus and the others to T. corymbosus. Within T. corymbosus, the different clades did not correlate with biome, geographical region, host, or any apparent morphological feature of the mistletoe. The morphologically distinct cactus parasite T. aphyllus likely arose in sympatry from an unspecialized tree parasite, T. corymbosus, after a host switch. The present day haplotype distribution is complex and resulted from post glaciation migrations from multiple Pleistocene refugia. PMID- 21628302 TI - Meta-analysis of cardiovascular disease risk markers in women with polycystic ovary syndrome. AB - BACKGROUND: The relation between polycystic ovary syndrome (PCOS) and cardiovascular disease (CVD) remains unclear. In an attempt to provide high quality evidence on the relation between PCOS and CVD, relevant literature for CVD risk markers [C-reactive protein (CRP), homocysteine (Hcy), tumor necrosis factor-alpha (TNF-alpha), plasminogen activator inhibitor-1 (PAI-1), lipoprotein (a) [Lp(a)], advanced glycation end-products (AGEs), vascular endothelial growth factor (VEGF), interleukin-6 (IL-6), asymmetric dimethylarginine (ADMA), endothelin-1 (ET-1) and fibrinogen] in women with PCOS was reviewed and analyzed. METHODS: A systematic search was conducted electronically using specific eligibility criteria. Weighted mean differences (WMDs) and 95% confidence intervals (CIs) were calculated and combined appropriately. To ensure synthesis of the best available evidence, sensitivity analyses were performed. RESULTS A total of 130 data sets were included in 11 different outcomes, involving 7174 and 5076 CVD markers in women with PCOS and controls, respectively. Women with PCOS demonstrated significantly elevated CRP [WMD (95% CI) 0.99 (0.77-1.21)], Hcy [2.25 (1.46-3.03)], PAI-1 antigen [16.96 (7.25-26.28)], PAI-1 activity [0.71 (0.18-1.23)], VEGF [1.72 (0.96-2.48)], ADMA [0.19 (0.08-0.3)], AGEs [3.91 (2.36 5.45)] and Lp(a) [0.81 (0.58-1.04)] concentrations compared with controls, yet with significant between-study heterogeneity. Borderline significance (not robust in the sensitivity analyses) was detected for TNF-alpha [0.75 (0.07-1.44)], ET-1 [1.06 (0.52-1.59)] and fibrinogen [0.20 (0.01-0.39)], whereas no difference was detected for IL-6 [0.71 (-0.16 to 1.59)]. CONCLUSIONS Women with PCOS have increased serum concentrations of CVD risk markers compared with controls. Whether this apparent risk is translated into increased incidence of CVD in later life remains to be elucidated. PMID- 21628303 TI - Molecular analysis of NDM-1-producing enterobacterial isolates from Geneva, Switzerland. AB - OBJECTIVES: To analyse the mechanisms responsible for decreased susceptibility or resistance to carbapenems in several enterobacterial isolates recovered in 2009 10 in Geneva University Hospitals, Switzerland. METHODS: PCR and sequencing were used to identify beta-lactamases, 16S RNA methylases and plasmid-mediated quinolone resistance genes. The transferable properties of the plasmids were analysed, as well as their plasmid type. The strains were typed by multilocus sequence typing. RESULTS: Three patients were found to be positive for NDM-1 producing enterobacterial isolates (one with Escherichia coli and Klebsiella pneumoniae, one with K. pneumoniae only and one with Proteus mirabilis), where NDM-1 stands for New Delhi metallo-beta-lactamase-1. The bla(NDM-1) carbapenemase gene was detected in all isolates in addition to genes encoding narrow-spectrum beta-lactamases (TEM-1, SHV-11, OXA-1, OXA-9 and OXA-10), extended-spectrum beta lactamases (CTX-M-15, CMY-16 and CMY-30), ArmA and quinolone resistance determinants (Qnr). The bla(NDM-1) gene was located on conjugative IncA/C- or IncF-type plasmids. Upstream of the bla(NDM-1) gene, part of ISAba125, previously identified in NDM-1-negative Acinetobacter baumannii, was found. Downstream of the bla(NDM-1) gene, variable sequences were found. CONCLUSIONS: This work constitutes the first identification of NDM-1 producers in Switzerland. Interestingly, patients from whom these NDM-1-producing isolates were recovered had a link with the Indian subcontinent or the Balkans. PMID- 21628304 TI - Trends of invasive serotype 6C pneumococci in Spain: emergence of a new lineage. AB - OBJECTIVES: To analyse the epidemiology of isolates of serotype 6C among invasive pneumococci isolated from children and adults in Spain between 1997 and 2009, and to characterize serotype 6C clones and macrolide and quinolone resistance mechanisms. METHODS: Antimicrobial susceptibility was determined following CLSI guidelines. Phenotypic characterization of macrolide-resistant isolates was performed by the double disc diffusion method. Genes associated with resistance to erythromycin and tetracycline were sought by PCR, while quinolone resistance was analysed by restriction fragment length polymorphism of the quinolone resistance-determining region. Isolates were typed by multilocus sequence typing. RESULTS: Seven hundred and eighty-nine of 866 serotype 6A pneumococci collected from 1997 to 2009 were available. Of these, 213 (27.0%) were serotype 6C; 16/163 (9.8%) in the 1997-2001 (pre-PCV7) period, 37/322 (11.5%) in the 2002-05 (early PCV7) period and 160/381 (42.0%) in the 2006-09 (late-PCV7) period. The overall proportions of serotype 6C increased from 0.1% (pre-PCV7) to 1% (late-PCV7) for paediatric isolates and from 0.3% to 1.7% among adult isolates. A major serotype 6C lineage (ST224/ST1150/ST4821), accounting for 66.7% of the isolates, was identified across the whole period. In the late-PCV7 period the antimicrobial non susceptibility of serotype 6C increased in association with the emergence of the ST386/ST4310/ST4825 lineage, which carried a Tn6002 transposon [erm(B) and tet(M) genes]. CONCLUSIONS: Serotype 6C pneumococci were identified in Spain during the period 1997-2009. The increase in serotype 6C in the late-PCV7 period was associated with the spread of the ST224/ST1150/ST4821 lineage and the emergence of the ST386/ST4310/ST4825 lineage. PMID- 21628305 TI - Cost-effectiveness analysis of anidulafungin versus fluconazole for the treatment of invasive candidiasis. AB - BACKGROUND: Anidulafungin was found to be non-inferior to and possibly more efficacious than fluconazole for treatment of invasive candidiasis (IC) in a major randomized clinical trial (RCT). There are no data comparing the cost effectiveness between azoles and echinocandins in treating IC. This economic analysis investigated the cost-effectiveness of anidulafungin compared with fluconazole for treatment of IC in an Australian setting. METHODS: A decision analytic model was constructed to capture downstream consequences of using either agent for treatment of IC. The main outcomes analysed in the model were treatment success and treatment failure (observed and indeterminate). Outcome probabilities and treatment pathways were derived from a published RCT. Resources used were estimated by an expert panel and cost inputs were derived from the latest Australian resources. The analysis was based on an Australian hospital perspective. Sensitivity analyses were conducted using Monte Carlo simulation. RESULTS: Anidulafungin (AU$74,587) had a higher total cost than fluconazole (AU$60,945) per successfully treated patient, primarily due to its higher acquisition cost. Hospitalization was the main cost driver for both comparators. However, when the rates of mortality in both treatment arms were considered, treatment with anidulafungin was expected to save an additional 0.53 life-years, with an incremental cost-effectiveness ratio (ICER) of AU$25 740 per life-years saved, which was below the implicit ICER threshold value for Australia. The results were robust over a wide range of variables. CONCLUSIONS: This is the first economic evaluation of anidulafungin versus fluconazole in the treatment of IC in Australia. Anidulafungin appears to be a cost-effective option. PMID- 21628306 TI - Efficacy and safety of long-term low molecular weight heparin in patients with antiphospholipid syndrome. AB - OBJECTIVE: Evaluation of the effectiveness and safety of long-term low molecular weight heparin (LMWH) in patients with antiphospholipid syndrome (APS) that had not previously responded to or tolerated oral vitamin K antagonists. METHODS: 23 patients with confirmed diagnosis of APS were retrospectively recruited. All patients were receiving LMWH as a result of intolerance and/or lack of response to warfarin therapy. The type of LMWH, the duration of treatment, the reason for switching to LMWH and the adverse effects were recorded. Outcomes were classified as no improvement, partial improvement or total improvement after at least 1 year of using LMWH. RESULTS: The average duration of LMWH treatment was 36 months. Most of the patients were on treatment with enoxaparin (n=16, 69%) and were switched to LMWH from warfarin mainly because of thrombosis despite therapeutic international normalised ratio (n=9, 39%). Good quality of life with no evidence of recurrent thrombotic events was noted in nine patients (39%), whereas 11 (48%) showed partial clinical improvement but no evidence of recurrent thrombotic episodes. Osteoporosis was reported in five patients (23%), all of whom were also receiving treatment with corticosteroids. CONCLUSIONS: Long-term LMWH may be a safe and effective alternative to warfarin for APS patients. PMID- 21628307 TI - Deletion of LCE3C_LCE3B is associated with rheumatoid arthritis and systemic lupus erythematosus in the Chinese Han population. AB - OBJECTIVES: The deletion of LCE3C_LCE3B confers susceptibility to psoriasis and rheumatoid arthritis (RA) in Caucasians. The aim of this study was to investigate the variant involvement in RA in the Chinese Han population and to further explore its potential role in the susceptibility to systemic lupus erythematosus (SLE). METHODS: LCE3C_LCE3B-del was genotyped in 898 patients with RA and 681 healthy controls. Two single nucleotide polymorphisms (SNPs, rs4112788 and rs4085613) in strong linkage disequilibrium with LCE3C_LCE3B-del were then genotyped in patients with RA (n=1222), SLE (n=870) and healthy controls (n=1031). RESULTS: The deletion of LCE3C_LCE3B and SNPs rs4112788 and rs4085613 showed an association with RA (allele analysis: p=7.72*10(-4), OR 1.28, 95% CI 1.11 to 1.47; p=6.39*10(-4), OR 1.23, 95% CI 1.09 to 1.38; and p=5.38*10(-4), OR 1.23, 95% CI 1.10 to 1.39, respectively). The two SNPs were also significantly associated with SLE (allele analysis: p=7.68*10(-3), OR 1.19, 95% CI 1.05 to 1.36 and p=5.30*10(-3), OR 1.20, 95% CI 1.06 to 1.37). CONCLUSIONS: This study provides evidence for an association between LCE3C_LCE3B-del and RA in non Caucasian populations, and SNPs rs4112788 and rs4085613 tagging LCE3C_LCE3B-del were novel susceptibility factors for SLE. PMID- 21628308 TI - Animal models for arthritis: innovative tools for prevention and treatment. AB - The development of novel treatments for rheumatoid arthritis (RA) requires the interplay between clinical observations and studies in animal models. Given the complex molecular pathogenesis and highly heterogeneous clinical picture of RA, there is an urgent need to dissect its multifactorial nature and to propose new strategies for preventive, early and curative treatments. Research on animal models has generated new knowledge on RA pathophysiology and aetiology and has provided highly successful paradigms for innovative drug development. Recent focus has shifted towards the discovery of novel biomarkers, with emphasis on presymptomatic and emerging stages of human RA, and towards addressing the pathophysiological mechanisms and subsequent efficacy of interventions that underlie different disease variants. Shifts in the current paradigms underlying RA pathogenesis have also led to increased demand for new (including humanised) animal models. There is therefore an urgent need to integrate the knowledge on human and animal models with the ultimate goal of creating a comprehensive 'pathogenesis map' that will guide alignment of existing and new animal models to the subset of disease they mimic. This requires full and standardised characterisation of all models at the genotypic, phenotypic and biomarker level, exploiting recent technological developments in 'omics' profiling and computational biology as well as state of the art bioimaging. Efficient integration and dissemination of information and resources as well as outreach to the public will be necessary to manage the plethora of data accumulated and to increase community awareness and support for innovative animal model research in rheumatology. PMID- 21628309 TI - Serum beta2 microglobulin correlates with the new ESSDAI in patients with Sjogren's syndrome. PMID- 21628310 TI - Determination of smoking status of rheumatoid arthritis patients by means of a self-reported questionnaire and serum cotinine assay: a cross-sectional study. PMID- 21628311 TI - Visualization of an anomalous left main coronary artery stenosis using transthoracic Doppler echocardiography. PMID- 21628312 TI - Increasing B-type natriuretic peptide levels predict mortality in unselected haemodialysis patients. AB - AIMS: Cardiac disease is the major cause of death in patients undergoing chronic haemodialysis. Recent studies have found that B-type natriuretic peptide (BNP) levels accurately reflect the cardiovascular burden of dialysis patients. However, the prognostic potential of BNP measurements in dialysis patients remains unknown. METHODS AND RESULTS: The study included 113 chronic dialysis patients who were prospectively followed up. Levels of BNP were measured at baseline and every 6 months thereafter. The potential of baseline BNP and annual BNP changes to predict all-cause and cardiac mortality were assessed as endpoints. Median follow-up was 735 (354-1459) days; 35 (31%) patients died, 17 (15%) of them from cardiac causes. Baseline BNP levels were similar among survivors and non-survivors, and failed to predict all-cause and cardiac death. Cardiac death was preceded by a marked increase in BNP levels. In survivors BNP levels remained stable [median change: +175% (+20-+384%) vs. -14% (-35-+35%) over the 18 months preceding either death or the end of follow-up, P< 0.001]. Hence, annual BNP changes adequately predicted all-cause and cardiac death in the subsequent year {AUC(all-cause) = 0.70 [SD 0.05, 95% CI (0.60-0.81)]; AUC(cardiac) = 0.82 [SD 0.04, 95%CI (0.73-0.90)]}. A BNP increase of 40% provided the best cut-off level. Cox regression analysis confirmed that annual increases over 40% were associated with a seven-fold increased risk for all-cause and cardiac death. CONCLUSIONS: Annual BNP increases above 40% predicted all-cause and cardiac death in the subsequent year. Hence, serially measuring BNP levels may present a novel tool for risk stratification and treatment guidance of end stage renal disease patients on chronic dialysis. PMID- 21628313 TI - In vitro methods for peptide display and their applications. AB - The presentation of recombinant peptide libraries linked to their coding sequence can be referred to as 'peptide display'. Phage display is the most widely practiced peptide display technology but more recent alternatives such as CIS display, ribosome display and mRNA display offer advantages over phage for speed, library size and the display of unnatural amino acids. These have provided researchers with tools to address some of the failings of peptides such as their low affinity, low stability and inability to cross biological membranes. In this review, we assess some of the recent advances in peptide display and its application. PMID- 21628314 TI - RNA splicing: disease and therapy. AB - The majority of human genes that encode proteins undergo alternative pre-mRNA splicing and mutations that affect splicing are more prevalent than previously thought. The mechanism of pre-mRNA splicing is highly complex, requiring multiple interactions between pre-mRNA, small nuclear ribonucleoproteins and splicing factor proteins. Regulation of this process is even more complicated, relying on loosely defined cis-acting regulatory sequence elements, trans-acting protein factors and cellular responses to varying environmental conditions. Many different human diseases can be caused by errors in RNA splicing or its regulation. Targeting aberrant RNA provides an opportunity to correct faulty splicing and potentially treat numerous genetic disorders. Antisense oligonucleotide therapies show particular promise in this area and, if coupled with improved delivery strategies, could open the door to a multitude of novel personalized therapies. PMID- 21628315 TI - From genotype to phenotype in bovine functional genomics. AB - In past 5 years, the promise that came with genome sequencing has revolutionized the functional genomics research field at unprecedented manner. It would soon know what all known genes do, particularly genes involved in genetic improvement of animal health and increase food animal production. With the availability of full bovine genomic sequence, yet we still have a lot of daunting tasks on 'genotype-to-phenotype problem' particularly about the phenotypic variations and trying to predict what genes are likely to be involved, and improved integrated interactive database. This article outlined and discussed about the current status of bovine functional genomics, recent development in bovine genome databases particularly in annotation of bovine genome, bovine quantitative trait loci database and its potential impact to unveil the from genotype-to-phenotype problem. PMID- 21628316 TI - Co-aggregate formation of CADASIL-mutant NOTCH3: a single-particle analysis. AB - CADASIL (cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy) is the most common monogenic cause of stroke and vascular dementia. Accumulation and deposition of the NOTCH3 (N3) extracellular domain in small blood vessels has been recognized as a central pathological feature of the disease. Recent experiments suggested enhanced formation of higher order multimers for mutant N3 compared with wild-type (WT). However, the mechanisms and consequences of N3 multimerization are still poorly understood, in part because of the lack of an appropriate in vitro aggregation assay. We therefore developed and validated a robust assay based on recombinant N3 fragments purified from cell culture supernatants. Using single-molecule analysis techniques such as scanning for intensely fluorescent targets and single-particle fluorescence resonance energy transfer, we show that spontaneous aggregation is limited to CADASIL mutant N3, recapitulating a central aspect of CADASIL pathology in vitro. N3 aggregation requires no co-factor and is facilitated by sulfhydryl crosslinking. Although WT N3 does not exhibit multimerization itself, it can participate in aggregates of mutant N3. Furthermore, we demonstrate that thrombospondin-2, a known interaction partner of N3, co-aggregates with mutant N3. Sequestration of WT N3 and other proteins into aggregates represents a potentially important disease mechanism. These findings in combination with a new assay for single molecule aggregation analysis provide novel opportunities for the development of therapeutic strategies. PMID- 21628317 TI - Sarcolemma instability during mechanical activity in Largemyd cardiac myocytes with loss of dystroglycan extracellular matrix receptor function. AB - The abnormal glycosylation and loss of extracellular matrix receptor function of the protein dystroglycan (DG) lead to the development of muscular dystrophy and cardiomyopathy. Dystroglycan is an important receptor for extracellular matrix proteins, such as laminin, in the basement membrane surrounding muscle. Large(myd) mice have a null mutation in a gene encoding the glycosyltransferase LARGE that results in abnormal glycosylation of alpha-DG and phenotypes similar to those in human alpha-DG glycosylation-deficient muscular dystrophy. Here, we show that Large(myd) hearts with the loss of DG extracellular matrix receptor function display a cardiomyopathy characterized by myocyte damage in patches of cells positive for membrane impermeant dyes. To examine the cellular mechanisms, we show that isolated adult cardiac myocytes from Large(myd) mice retain normal laminin-dependent cell adhesion, cell surface laminin deposition and basement membrane assembly. However, although isolated adult cardiac myocytes with the loss of alpha-DG glycosylation adhere normally to laminin substrates both passively and in the presence of mechanical activity, Large(myd) myocytes rapidly take up membrane impermeant dye following cyclical cell stretching. Therefore, while other cell surface laminin receptors are likely responsible for myocardial cell adhesion to the basement membrane, DG has a unique function of stabilizing the cardiac myocyte plasma membrane during repetitive mechanical activity by tightly binding the transmembrane dystrophin-glycoprotein complex to the extracellular matrix. This function of DG to stabilize the myocyte membrane during normal physiologic cell length changes is likely critical for the prevention of the myocardial damage and subsequent remodeling observed in alpha DG glycosylation-deficient muscular dystrophies. PMID- 21628318 TI - Delayed sternal closure after vacuum-assisted closure therapy for tracheo innominate artery fistula repair. AB - We report a case of successful innominate artery resection with delayed sternal closure after vacuum-assisted closure (VAC) therapy for a tracheo-innominate artery fistula (TIF). A 42-year-old woman with cerebral palsy underwent tracheostomy for respiratory assistance. On postoperative day 14, she was transferred to our hospital after an episode of massive hemoptysis. TIF was diagnosed based on the findings of multidetector computed tomography. Thus, we resected the innominate artery and started VAC therapy to control the postoperative local infection. The patient recovered uneventfully, without any infectious sequelae. Our strategy, which includes VAC therapy, for TIF repair may eliminate postoperative infective problems that could induce sequential bleeding and sternal compromise. To our knowledge, this is the first report of using VAC therapy for TIF. PMID- 21628319 TI - Discordance of aortic remodeling with clinical outcomes in patients treated with endovascular repair for uncomplicated type B aortic dissection. AB - Uncomplicated type B aortic dissection is managed with anti-impulse therapy since surgery offers no additional long-term survival advantage. In recent years, thoracic endovascular repair (TEVAR) has been forwarded as a treatment strategy that may retard aortic growth, lower rupture risk, and improve clinical outcome compared to medical management in patients with uncomplicated type B aortic dissection. Although aortic remodeling often serves as a surrogate measure of treatment success in trials of aortic dissection, there is, in fact, little evidence to suggest that aortic remodeling confers a clinical advantage in this patient cohort. TEVAR likely will not be widely recommended for the patient with uncomplicated type B aortic dissection until a prospective comparative clinical trial demonstrates a clear clinical advantage of TEVAR over medical management. Measures of aortic remodeling are poor surrogate measures of treatment success in this patient population. PMID- 21628320 TI - Endovascular stent grafting of a complicated type B aortic dissection with approach through the innominate artery using a side-branch graft. AB - Endovascular stent grafting has emerged as an effective method for the treatment of complicated acute type B aortic dissections. In cases where access to the aorta cannot be established through the ileofemoral trunks, the axillary arteries are used as an alternative route. Often, however, these arteries are too small to accommodate the device sheath. We report the case of an 82-year-old female with complicated type B aortic dissection treated with deployment of an endovascular stent-graft through an 8 mm Dacron graft sewn to the innominate artery. PMID- 21628321 TI - Complete excision of giant calcified saphenous vein graft aneurysm in redo coronary artery bypass grafting. AB - A 73-year-old female who first underwent coronary artery bypass graft surgery in 1987 represented in 2008 with recurrent angina. Coronary angiography demonstrated a giant calcified aneurysm of the saphenous vein graft (SVG) to the right coronary artery bracketed by severe stenoses in addition to severe disease in the native vessels. Following 64-slice computed tomography coronary angiography, total resection of the aneurysm with en-bloc excision of the contagious right atrial free wall and ascending aorta were accomplished during redo coronary revascularization. The pathophysiology of SVG aneurysm, the clinical relevance and therapeutic approaches will be discussed in light of this case. PMID- 21628322 TI - Pseudoaneurysm of mitral-aortic intervalvular fibrosa. AB - The mitral-aortic intervalvular fibrosa (MAIF) is involved in the functional integrity of the two valves. We discuss the clinical decision-making in a population with MAIF pseudoaneurysms following valve replacement surgery. From 2003 to 2010, 16 patients (age 65 +/- 17 years) were referred to our institute with worsening valvular disease. All the patients had undergone previous aortic or mitral valve replacement surgery. Transthoracic and transesophageal echocardiography was performed in these patients. According to the echocardiographic findings, we described two groups of patients with MAIF pseudoaneurysms: group 1 (seven patients) without previously known infective disease, and group 2 (nine patients) with a medical history of endocarditis. For asymptomatic patients without an infective background or valve regurgitation (group 1), clinical and echocardiographic follow-up was selected. Reintervention was planned for postinfection pseudoaneurysms (group 2), on account of the residual paravalvular leak and the high-risk of MAIF rupture. The case series emphasizes the need for more awareness about MAIF pseudoaneurysms, rare complications of aortic and mitral valve surgery, which occur far more frequently than expected. Although clinical decision-making is debatable, it may be helpful to differentiate patients into those with or without previous endocarditis, and according to heart failure symptoms. PMID- 21628323 TI - Half-turned truncal switch operation for transposition of great arteries with ventricular septal defect and pulmonary regurgitation. AB - A three-month-old girl weighing 4.2 kg, diagnosed with transposition of the great arteries (TGA) and ventricular septal defect (VSD) was referred to us. She had normal-sized pulmonary annulus and moderate pulmonary regurgitation. Because her pulmonary valve was not suitable for systemic circulation due to valvular incompetence, the half-turned truncal switch operation was selected. The postoperative course was uneventful without left or right ventricular outflow obstructions over a year of follow-up. Our report demonstrated that the TGA and VSD with normal pulmonary annulus is not contraindicated for half-turned truncal switch operation. PMID- 21628324 TI - Primary pulmonary myoepithelial carcinoma. AB - Primary salivary gland-type tumors of the lung are rare neoplasms. The pulmonary myoepithelial carcinoma belongs to this group. Since it was first described in 1998, we have identified only seven actual cases reported in the literature so far. We describe the case of a non-smoker 60-year-old Caucasian female, who was referred to our institution for the treatment of three peripheral nodules. Her medical history revealed that a wedge resection, for pulmonary myoepithelial carcinoma, was performed in another center 15 months previously. After resection of the current nodules, the histological findings showed three myoepithelial carcinomas of the lung. There was no vascular or lymphatic invasion. The hilar and mediastinal lymph nodes were negatives. The patient is doing well without any sign of recurrence. Herein we shall discuss the case of a primary pulmonary myoepithelial carcinoma in a female and propose a brief review of the literature. PMID- 21628326 TI - FoxO3 induces reversible cardiac atrophy and autophagy in a transgenic mouse model. AB - AIMS: The transcription factor FoxO3 contributes to anti-hypertrophic signalling in the heart presumably by regulating autophagic-lysosomal and ubiquitin proteasomal pathways. We wanted to study FoxO3 function in the adult heart in vivo by expressing a constitutively active mutant of FoxO3 in transgenic mice. METHODS AND RESULTS: We generated transgenic mice in which a tetracycline regulated constitutively active FoxO3 transgene (FoxO3-CA) is controlled by the heart-specific alpha-myosin heavy chain promoter. Cardiac-specific expression in adult mice resulted in a decrease in heart weight by 25% and a reduction in stroke volume and cardiac output. The decrease in heart size was due to a reduction in the size of individual cardiomyocytes, whereas there was no evidence for increased cell death. FoxO3 activation was accompanied by the initiation of a foetal gene programme with increased expression of beta-myosin heavy chain and natriuretic peptides, and by the activation of AKT and mammalian target of rapamycin signalling. As shown by electron microscopy, FoxO3-CA massively stimulated destruction of sarcomeres and autophagy, and induced expression of LC3 II and BNIP3. When FoxO3-CA expression was shut off in affected mice, cardiac atrophy and dysfunction as well as molecular markers were normalized within 1 month. FoxO3-CA expression did not counteract hypertrophy induced by transverse aortic constriction. CONCLUSION: Heart-specific expression of constitutively active FoxO3 leads to reversible heart atrophy. The reversibility of the phenotype suggests a remarkable ability of the adult myocardium to respond to different regulatory cues. PMID- 21628327 TI - Anaphylactoid reaction to facial adder bite. PMID- 21628328 TI - Purple glove syndrome following intravenous phenytoin administration. PMID- 21628329 TI - The historical milestones in the understanding of leukocyte biology initiated by Elie Metchnikoff. AB - Progress in science is made with key discoveries, correct analyses, wrong statements, and disputes within the scientific community. Despite scientific controversies, Elie Metchnikoff has allowed the theory of phagocytes to triumph. Starting his career as a zoologist, Metchnikoff became a pathologist, beautifully defining the role of monocytes, macrophages, and neutrophils during inflammation and innate immunity. The discoveries of immune cells were made by other outstandings scientists, such as Paul Ehrlich, whose key contributions to humoral immunity led him to share the Nobel Prize with Metchnikoff. Ludwig Aschoff grouped certain cells under the term RES, according to their propensity for absorbing and storing vital stains. This classification was not always a source of accurate discoveries, and research on the exact function of RES cells led to some wanderings. This is illustrated by studies about the nature of the antibody producing cells, which were first thought to belong to the RES, before being identified as plasmocytes and lymphocytes. PMID- 21628330 TI - Improved host defense against septic peritonitis in mice lacking MyD88 and TRIF is linked to a normal interferon response. AB - The signaling adapters MyD88 and TRIF are engaged by TLRs and/or receptors of the IL-1 family and are considered important for innate immune responses that combat bacterial infections. Here, the consequences of a combined MyD88 and TRIF deficiency for the innate immune response against severe septic peritonitis was examined. We demonstrate that Myd88(-/-);Trif(Lps2/Lps2) mice had markedly reduced bacterial numbers in the peritoneal cavity and peripheral blood, indicating that bacterial clearance in this model is improved in the absence of MyD88/TRIF signals. Survival of Myd88(-/-); Trif(Lps2/Lps2) mice was improved significantly. The lack of MyD88/TRIF signaling prevented the excessive production of inflammatory cytokines and of IL-10. In contrast, Ifng mRNA was expressed at WT levels, and induction of Ifnb mRNA was reduced only by one-half. Consistent with these findings, numerous IFN-regulated genes, including p47 and p65 GTPases, as well as CXCL10, were expressed in a MyD88/TRIF-independent manner. In support of the in vivo data, Myd88(-/-); Trif(Lps2/Lps2) macrophages stimulated with live intestinal bacteria produced normal amounts of CXCL10. The production of p47 GTPases and CXCL10 in septic peritonitis was found to be dependent on the presence of IFNAR1, but not IFN-gamma, indicating a normal induction of the type I IFN response in Myd88(-/-);Trif(Lps2/Lps2) mice, despite attenuated IFN-beta production. Together, these results provide evidence that in severe septic peritonitis, the absence of MyD88 and TRIF balances the innate immune response in a favorable manner by attenuating deleterious responses such as excessive cytokine release, while leaving intact protective IFN responses. PMID- 21628331 TI - Gene expression analysis of dendritic cells that prevent diabetes in NOD mice: analysis of chemokines and costimulatory molecules. AB - We have demonstrated previously that BM-derived DCs can prevent diabetes development and halt progression of insulitis in NOD mice, the mouse model of type 1 diabetes. The DC population that was most effective in this therapy had a mature phenotype, expressed high levels of costimulatory molecules, and secreted low levels of IL-12p70. The protective DC therapy induced Treg and Th2 cells in vitro and in vivo. Microarray analysis of therapeutic and nontherapeutic DC populations revealed differences in the expression of OX40L, CD200, Ym-1, CCL2, and CCL5, which could play important roles in the observed DC-mediated therapy. The unique pattern of costimulatory molecules and chemokines expressed by the therapeutic DCs was confirmed by flow cytometry and ELISA. Using a novel cell labeling and (19)F NMR, we observed that the chemokines secreted by the therapeutic DCs altered the migration of diabetogenic Th1 cells in vivo and attracted Th2 cells. These results suggest that the therapeutic function of DCs is mediated by a combination of costimulatory and chemokine properties that results in the attraction of diabetogenic Th1 and the induction of Th2 and/or Treg differentiation. PMID- 21628332 TI - Suppressor of cytokine signaling (SOCS)1 is a key determinant of differential macrophage activation and function. AB - Macrophages become activated by their environment and develop polarized functions: classically activated (M1) macrophages eliminate pathogens but can cause tissue injury, whereas alternatively activated (M2) macrophages promote healing and repair. Mechanisms directing polarized activation, especially in vivo, are not understood completely, and here, we examined the role of SOCS proteins. M2 macrophages activated in vitro or elicited by implanting mice i.p. with the parasitic nematode Brugia malayi display a selective and IL-4-dependent up-regulation of SOCS1 but not SOCS3. Using siRNA-targeted knockdown in BMDM, we reveal that the enhanced SOCS1 is crucial for IL-4-induced M2 characteristics, including a high arginase I:iNOS activity ratio, suppression of T cell proliferation, attenuated responses to IFN-gamma/LPS, and curtailed SOCS3 expression. Importantly, SOCS1 was essential in sustaining the enhanced PI3K activity that drives M2 activation, defining a new regulatory mechanism by which SOCS1 controls M2 polarization. By contrast, for M1 macrophages, SOCS1 was not only an important regulator of proinflammatory mediators (IL-6, IL-12, MHC class II, NO), but critically, for M1, we show that SOCS1 also restricted IL-10 secretion and arginase I activity, which otherwise would limit the efficiency of M1 macrophage proinflammatory responses. Together, our results uncover SOCS1, not only as a feedback inhibitor of inflammation but also as a critical molecular switch that tunes key signaling pathways to effectively program different sides of the macrophage balance. PMID- 21628333 TI - Reciprocal regulation of CXCR4 and CXCR7 in intestinal mucosal homeostasis and inflammatory bowel disease. AB - IBDs are characterized by increased influx of immune cells to the mucosa of genetically susceptible persons. Cellular migration to injury sites is mediated by chemokines. CXCL12 is a ubiquitous, constitutive chemokine that participates in stem cell proliferation and migration and mediates T lymphocyte migration to inflamed tissues. We have recently reported that CXCL12 and its receptor, CXCR4, are expressed in normal and more prominently, inflamed human intestinal mucosa. However, the interactions and roles of CXCL12 and its receptors, CXCR4 and the recently discovered CXCR7, in intestinal inflammation have not been defined. In the present study, we further dissected the effects of CXCL12 on lymphocytes in intestinal homeostasis and inflammation and delineated the interplay between CXCL12 and its receptors CXCR4 and CXCR7. To that end, fresh mononuclear cells were isolated from mucosa and PB of healthy or IBD patients. Phenotypical and functional assays were conducted using flow cytometry, Transwell migration chambers, and ELISA. The data show that CXCL12-mediated migration of T cells is CXCR4- but not CXCR7-dependent. T cell activation reciprocally regulates CXCR7 and CXCR4 expression and migratory capacity. IBD PBTs expressed more CXCR7 than normal PBTs. Finally, T cells attracted by CXCL12 are mostly of a memory phenotype. In conclusion, the present study suggests that the interplay between CXCL12 and its receptors affects homeostasis and inflammation in the intestinal mucosa. PMID- 21628334 TI - Editorial: Interfering with B cell immunity. PMID- 21628335 TI - Editorial: Cytohesin-1, on the tail of two integrins. PMID- 21628336 TI - The social justice roots of the Mentors in Violence Prevention model and its application in a high school setting. AB - The social justice roots and theory of the Mentors in Violence Prevention (MVP) model is presented, followed by an empirical study examining the influence of MVP in high school settings. Findings reveal students exposed to the MVP model are more likely to see forms of violence as being wrong and are more likely to take actions to intervene than students not exposed to the program. Findings support the premises on which MVP is founded. PMID- 21628337 TI - Sisterhood may be powerful for reducing sexual and intimate partner violence: an evaluation of the Bringing in the Bystander in-person program with sorority members. AB - Sorority members may be at greater risk than other college women for sexual violence and intimate partner violence (IPV). We evaluated the Bringing in the Bystander in-person program with sorority members who participated in the program (n = 30) compared with those who did not (n = 18). Results indicate that program participants showed increased bystander efficacy, likelihood to help, and responsibility for ending violence without unintended "backlash" effects. Implications include a call for future programming with more diverse sorority members over longer time. In addition, we discuss what the findings might mean for formal campus policies and practices for preventing sexual violence and IPV. PMID- 21628338 TI - Rehearsing for real life: the impact of the InterACT Sexual Assault Prevention Program on self-reported likelihood of engaging in bystander interventions. AB - The interACT Sexual Assault Prevention Program is an interactive, skill-building performance based on the pedagogy of Augusto Boal's Theatre of the Oppressed. A longitudinal evaluation of this program compared pretest, posttest, and 3-month follow-up data from 509 university student participants. Results suggested that the interACT performance was successful in increasing participants' beliefs about the effectiveness of bystander interventions and the self-rated likelihood that participants would engage in bystander interventions in the future. Differences in both overall ratings and rates of change were noted. Implications of these results for research and practice are discussed. PMID- 21628339 TI - Graft patency and late outcomes for patients with ST-segment elevation myocardial infarction who underwent coronary surgery. AB - OBJECTIVE: The aim of our study was to assess the long-term clinical outcomes and the grafts patency rates of patients with ST-segment elevation myocardial infarction (STEMI) who underwent urgent or emergency coronary artery bypass grafting (CABG). MATERIALS: Participants in two previous studies comprising 207 STEMI patients undergoing on-pump (145 patients) or off-pump (62 patients) coronary artery bypass graft (CABG) surgery in our institution were prospectively followed to assess late mortality, graft patency, and major adverse cardiac related event (MACE) rates. Graft patency was evaluated by multi-detector computed tomography angiography 64-slice scan. Mean times of graft implantation were 38+/-16 months and 37+/-14 months in on-pump and off-pump, respectively. Follow-up data were obtained in all patients and was 100% complete. RESULTS: Late mortality rate was 7.4% (10 patients) in the on-pump and 6.5% (4 patients) in off pump groups (p=0.45). Five-year overall survival rate (+/-SE) was 93.5+/-2.1% and 92.6+/-1.9% in the off-pump vs on-pump, respectively. Five years' freedom from cardiac-related death was 94.9+/-2.9% in the on-pump group vs 96.8+/-3.2% in the off-pump group (p=0.25). Five years' freedom from cardiac-related events was 89.7+/-1.6% in the on-pump group versus 93.5+/-1.8% in the off-pump group (p=0.32). In all patients, a total of 449/491 (91.5%) grafts were patent. Percentages of overall grafts classified as patent were similar in the on-pump group (90.7% - 322/355 conduits) versus the off-pump group (91% - 133/146 conduits). Graft patency rates were also similar between the two groups with regard to arterial and saphenous vein conduits, and with regard to different branches of the coronary arteries grafted. CONCLUSION: Our data suggest that off pump CABG patients have the same late mortality, MACEs, and graft patency rates as conventional cardioplegic cardiac arrest CABG patients. In our opinion, urgent or emergency CABG for patients with STEMI can be done either way. PMID- 21628340 TI - Is 5-fluorouracil-induced vasospasm a Kounis syndrome? A diagnostic challenge. AB - Cardiovascular hypersensitivity is a rare and well-documented side-effect of 5-FU (5-fluorouracil). Besides the common complications such as angina pectoris and myocardial infarction, it can also cause cardiogenic shock, and supraventricular and ventricular arrhythmias. Studies have reported that FU-induced angina most commonly occurred due to vasospasm. In our case, 9 hours after stopping the infusion of 5-FU, the patients developed symptoms and electrocardiographic (ECG) findings consistent with acute myocardial infarction. We intend to share this rare case and discuss whether this late complication after 5-FU infusion is an FU induced vasospasm or rather an allergic reaction leading to Kounis syndrome. PMID- 21628341 TI - Pharmacokinetics of butorphanol in broiler chickens. AB - Butorphanol tartrate (2 mg/kg) was injected intravenously in 18 healthy broiler chickens to study its pharmacokinetics. Plasma samples were analysed by a highly sensitive high-performance liquid chromatography with diode-array detection method and pharmacokinetic parameters were calculated from the mean pooled data. With non-compartmental analysis, the terminal half-life (T(1/2.z)) was 71.3 minutes, clearance was 67.6 ml/minute/kg and the apparent volume of distribution was 6.9 l/kg. The concentration-time curve was also fitted to a two-compartmental model. In this analysis, elimination half-life (T(1/2beta)) was 69.3 minutes, clearance was 74.6 ml/minute/kg and volume of distribution at steady state was 5.6 l/kg. The micro rate constants k(21), k(12) and k(10) were 0.034, 0.050 and 0.029, respectively. Butorphanol was well distributed in the chickens with rapid clearance. It remained above the minimum effective concentration for analgesia in mammals for approximately two hours in the chickens. PMID- 21628342 TI - Adolescent parricide and psychopathy. AB - Most cases of juvenile parricide are believed to be the result of child abuse, yet the vast majority of abused children do not kill their parental abusers. This study explored the role of psychopathy in 10 adolescent parricide offenders tried in adult court who were referred for pretrial psychiatric evaluation. In addition, psychopathological findings, crime-related behaviors, and judicial outcomes are described. Diagnostic and Statistical Manual of Mental Disorders diagnoses, most commonly posttraumatic stress disorder, and chronic, severe child abuse were prevalent. Psychopathic traits were not found to have played a role in the traumatized youths' parricidal behavior. Killings occurred in the family homes, usually through a surprise attack with parent-owned firearms. There was an average of 1.7 victims per event, with fathers being the most likely victim. Bodies were commonly covered or wrapped and moved postmortem. Confessions were often incredible in quality. One half received sentences of 40+ years, and the modal sanction was a life sentence, despite 90% premorbidly described as good youth. Further studies of adolescent parricide are needed to better understand this unusual population. PMID- 21628343 TI - Norepinephrine transporter -3081(A/T) and alpha-2A-adrenergic receptor MspI polymorphisms are associated with cardiovascular side effects of OROS methylphenidate treatment. AB - The purpose of this study was to investigate a possible association between norepinephrine genes and cardiovascular side effects of the Osmotic Controlled Release Oral Delivery System-methylphenidate (OROS-MPH) in Korean children with attention-deficit/hyperactivity disorder (ADHD). One hundred and one children with ADHD (8.7 +/- 1.7 years) were recruited from child psychiatric centers at six university hospitals in South Korea. All participants were drug-naive ADHD children treated with OROS-MPH for 12 weeks. During the treatment period the investigators titrated the OROS-MPH dosage on the basis of symptom severity and side effects. Resting heart rate (HR), diastolic blood pressure (DBP), and systolic blood pressure (SBP) were examined before and after treatment. The percentage change score (post-treatment - pretreatment/pretreatment * 100) of each parameter was calculated. Genotyping of SLC6A2 -3081(A/T) and G1287A, and alpha-2A-adrenergic receptor (ADRA2A) MspI and DraI polymorphisms was performed. Clinically significant changes were not found in cardiovascular monitoring during the course of treatment. An increase of HR after OROS-MPH treatment was found to be statistically significant (t = 3.54, p = 0.001). Changes in SBP and DBP were not significant and no specific change was found in the ECGs. However, an additive regression analysis demonstrated a significant association between SLC6A2 -3081(A/T) and percentage change in HR post-treatment (p = 0.01) after controlling for age, gender, dosage of MPH and response and baseline pulse rate. Children with ADHD having the T/T genotype of SLC6A2 showed a 12.5% increase in HR compared to baseline, whereas children with the A/T or A/A genotype showed a 3.5% and 2.5% increase after OROS-MPH treatment, respectively. There was also a significant association between the ADRA2A MspI genotype and percentage change of DBP post-treatment after controlling for age, gender, dosage of MPH and response and baseline DBP (p = 0.009). Children with ADHD having the C/C genotype of ADRA2A MspI showed an 18.5% increase in DBP compared to baseline, but children with the G/G or G/C genotype showed a 0.2% decrease after OROS-MPH treatment. The overall cardiovascular effects of OROS-MPH were modest. However, our findings show a positive association between norepinephrine-related gene polymorphisms and cardiovascular response induced by MPH in Korean children with ADHD. Consideration must be given to such children or adults with specific norepinephrine-related genotypes, especially if they show significant changes in HR or DBP after OROS-MPH administration. PMID- 21628344 TI - D1 and D2 dopaminergic systems in the rat basolateral amygdala are involved in anxiogenic-like effects induced by histamine. AB - Involvement of the dopamine receptors in the basolateral amygdala (BLA) in the effects of histamine on anxiety-like behaviors of the elevated plus maze in male Wistar rats was investigated. The results showed that bilateral intra-BLA injections of histamine (2.5, 5 and 7.5 ug/rat) induced an anxiogenic-like effect, revealed by decreases in percentage of open arm time (%OAT) and open arm entries (%OAE). Intra-BLA administration of dopamine D1 receptor agonist, SKF38393 (0.25 ug/rat), and dopamine D2 receptor agonist, quinpirole (0.03 and 0.05 ug/rat), decreased %OAT but not %OAE. Conversely, intra-BLA administration of dopamine D1 receptor antagonist, SCH23390 (0.5 and 1 ug/rat), and dopamine D2 receptor antagonist, sulpiride (0.3 and 0.5 ug/rat), increased %OAT and %OAE, suggesting an anxiolytic-like effect for both drugs. Interestingly, co administration of a silent dose of SCH23390 or sulpiride prevented anxiogenic like effects of SKF38393 and quinpirole, respectively. Conjoint administration of a sub-effective dose of SKF38393 (0.125 ug/rat) or quinpirole (0.01 ug/rat) along with lower doses of histamine (1 and 2.5 ug/rat) induced anxiolytic-like effects. On the other hand, intra-BLA pretreatment with a silent dose of SCH23390 (0.25 ug/rat) or sulpiride (0.1 ug/rat) prevented the anxiogenic-like effect of higher doses of histamine (5 and 7.5 ug/rat). No significant change was observed in total closed arm entries, as an index for motor activity of the animals. It can be concluded that the dopamine D1 and D2 receptors in the BLA may be involved in the anxiogenic-like effects induced by histamine. PMID- 21628345 TI - On site domestic organic waste treatment through vermitechnology using indigenous earthworm species. AB - In India the exotic epigeic species, Eisenia fetida is mostly used for vermicomposting. The introduction of exotic species into local bio system may affect the indigenous earthworm species population. A comparative study between exotic species (Eisenia fetida) and indigenous species Perionyx sansibaricus and Perionyx excavates was performed to determine the potential of indigenous species of the area vis-a-vis the exotic species for composting of domestic organic waste blended with cattle manure. The results of the study show a significant reduction in initial C/N ratio from 55 to 13 for P. excavates and 15 for P. sansibaricus of the ready product which was within the agronomic acceptable limit ( < 20). The total organic matter reduced by 50% and pH also reduced to be nearer to neutral, but there was an increase in total nitrogen to 102% and total phosphorus increased from the initial concentration of 7.62 g kg(-1) up to 13.2 g kg(-1). Overall, by employing above indigenous species, domestic organic waste can be directly converted into high-quality hygienic stable fertilizer (vermicompost) which is rich in nitrogen, phosphorus and potassium and free from pathogens. PMID- 21628346 TI - Study on thermal co-pyrolysis of jatropha deoiled cake and polyolefins. AB - Three plastics, high density polyethylene (HDPE), polypropylene (PP) and polystyrene (PS), were individually co-pyrolysed with deoiled cake of jatropha (JC) at 400 and 450 degrees C in a batch reactor in the presence of nitrogen under atmospheric pressure to produce modified liquid fractions. At higher temperature (450 degrees C), the yield of liquid fractions by the pyrolysis of plastics (HDPE, PP and PS) alone was found to increase by 11, 12.5 and 11% for HDPE, PP and PS, respectively. Furthermore, the gaseous fraction increased by 1.3 to 2.6% while the residue generation reduced by 12.3 to 15.1%. In comparison with only plastics pyrolysis, the yield of the liquid fraction improved by 2.0 to 4.9% for their co-pyrolysis with JC. Gas chromatography-mass spectrometry analyses demonstrated that the co-processing afforded a reduction of paraffin and olefins in the liquid fractions for all of the experiments. This reduction was found to be in the order of PS > PP > HDPE. Furthermore, the proportion of oxygenates in the liquid product increased in the order of PP > HDPE > PS. Physical characteristics such as oxygenates, water contents, acid values and viscosity increased during the co-pyrolysis of plastics and JC in contrast to the liquid fractions obtained from the pyrolysis of pure plastics. Furthermore, co-pyrolysis offered a reduction in calorific values. PMID- 21628347 TI - Characteristics of Kawasaki disease in older children. AB - OBJECTIVES: To evaluate characteristics of Kawasaki disease(KD) in older children in comparison with younger patients with the disease, and to improve the knowledge of clinicians on KD in older children. METHODOLOGY: All children with a discharge diagnosis of KD at Wuhan Union Hospital from January, 2004 to May, 2010 were retrospectively reviewed. RESULTS: A total of 113 patients were included in this study; 20 patients (17.7%) were >=5 years old at the time of illness and 93 patients (82.3%) were <5 years old. The older children seemed to have longer total fever duration, pre-IVIG and post-IVIG fever duration than the younger children. The individual clinical criteria appeared later in the course of KD in older children than in the younger ones except cervical lymphadenopathy. The older age group had a higher incidence of cervical lymphadenopathy in comparison with the younger group (85.0% vs 51.6%). ESR was quite different between the older and younger groups (85.92 vs 67.27). Overall the older age group had a higher prevalence of additional IVIG treatment and coronary artery abnormalities (60.0% vs 32.2%) versus the younger age group. CONCLUSION: For some reasons, KD in older children was difficult in early diagnosis and treatment. Also, older children may have a more marked inflammatory response and those treated with IVIG were more likely to require repeated IVIG treatment. And probably because of all these, older patients with KD had a higher prevalence of coronary artery abnormalities than the younger patients. PMID- 21628348 TI - The maintenance of certification conundrum: education-- education--education 2011. PMID- 21628349 TI - Early marriage, poor reproductive health status of mother and child well-being in India. AB - BACKGROUND AND METHODOLOGY: Early marriage, women's poor reproductive health and child well-being are important areas of concern, especially in developing countries like India. Data from the third wave of National Family Health Survey (NFHS, 2005-2006) was used to examine the effects of early marriage on the reproductive health status of women and on the well-being of their children. Bivariate analyses, multiple linear regression and logistic regression were used for analyses. RESULTS: The results show that early age at marriage had detrimental effects on the reproductive health status of women. Women married at an early age were exposed to frequent childbearing, unplanned motherhood and abortions, which negatively affected their nutritional status. Children born to mothers with poor reproductive health had lower chances of survival and a higher likelihood of anthropometric failure (i.e. stunting, wasting and underweight). DISCUSSION AND CONCLUSIONS: Programmes should focus on delaying entry of adolescents into wedlock and motherhood through information, education and communication. More emphasis needs to be put on meeting the reproductive needs of poor adolescent mothers, and improving the nutritional status of their children, to break the vicious circle of poor reproductive health and poverty. PMID- 21628350 TI - Thirty years of HIV and AIDS: future challenges and opportunities. AB - As the third decade since AIDS was first recognized comes to an end, extraordinary advances have occurred in the understanding, treatment, and prevention of HIV infection and AIDS. As a result of these successes, it is now time to focus on future challenges. Paramount among these is reaching the goal of truly controlling and ultimately ending the HIV and AIDS pandemic. To that end, AIDS researchers and public health personnel worldwide are aggressively pursuing 3 key areas of scientific research. Given the availability of highly effective therapeutic regimens for HIV infection, the first challenge is efficiently identifying a maximum number of HIV-infected persons through voluntary HIV testing and initiating antiretroviral therapy (ART). Second, scientists are trying to develop a cure for HIV infection, which would alleviate the need for lifelong ART. Finally, preventing new cases of HIV infection, which currently number approximately 2.6 million per year globally, is critical to any attempt to end this pandemic. This article addresses each of these challenges and provides directions for the future. PMID- 21628351 TI - Balancing punishment and compassion for seriously ill prisoners. AB - Compassionate release is a program that allows some eligible, seriously ill prisoners to die outside of prison before sentence completion. It became a matter of federal statute in 1984 and has been adopted by most U.S. prison jurisdictions. Incarceration is justified on 4 principles: retribution, rehabilitation, deterrence, and incapacitation. Compassionate release derives from the theory that changes in health status may affect these principles and thus alter justification for incarceration and sentence completion. The medical profession is intricately involved in this process because eligibility for consideration for compassionate release is generally based on medical evidence. Many policy experts are calling for broader use of compassionate release because of many factors, such as an aging prison population, overcrowding, the increasing deaths in custody, and the soaring medical costs of the criminal justice system. Even so, the medical eligibility criteria of many compassionate-release guidelines--which often assume a definitive prognosis--are clinically flawed, and procedural barriers may further limit their rational application. We propose changes to address these flaws. PMID- 21628352 TI - Midportion Achilles tendinosis and the plantaris tendon. AB - OBJECTIVES: When re-operating patients with midportion Achilles tendinosis, having had a poor effect of ultrasound (US) and Doppler-guided scraping, the author found the involvement of the plantaris tendon to be a likely reason for the poor result. The aim of this study was to investigate the occurrence of a plantaris tendon in close relation to the Achilles tendon in consecutive patients with midportion Achilles tendinosis undergoing treatment with US and Doppler guided scraping. MATERIAL AND METHODS: This study includes 73 consecutive tendons with chronic painful midportion Achilles tendinosis, where US+Doppler examination showed thickening, irregular tendon structure, hypo-echoic regions, and localised high blood flow outside and inside the ventral Achilles midportion. The tendons were treated with US+Doppler-guided scraping, via a medial incision. If there was a plantaris tendon located in close relation to the medial Achilles, it was extirpated. RESULTS: An invaginated, or 'close by located', enlarged plantaris tendon was found in 58 of 73 (80%) tendons. Preliminary clinical results of the combined procedure, US + Doppler-guided surgical scraping and extirpation of the plantaris tendon, are very promising. CONCLUSIONS: A thickened plantaris tendon located in close relation to the medial Achilles seems common in patients with chronic painful midportion tendinosis. The role of the plantaris tendon in midportion Achilles tendinosis needs to be further evaluated and should be kept in mind when treating this condition. PMID- 21628353 TI - Hypoglycaemia is more frequent in type 2 diabetic patients with co-morbid vascular disease: an analysis of the DiaRegis registry. AB - BACKGROUND: Patients with type-2 diabetes are at risk for treatment- and disease related complications. Little is known about the interrelation of hypoglycaemia and co-morbid vascular disease (VD), defined as coronary heart disease, stroke and peripheral arterial disease. HYPOTHESIS: Hypoglycaemia is associated with co morbid VD in diabetic patients. METHODS: DiaRegis is a prospective registry that included patients with type-2 diabetes in 2009/2010. Metric variables are displayed as median and quartiles. For the comparison of patients with or without VD Odds Ratios (OR) were determined from univariate analyses and adjusted for differences in patient characteristics (multivariable analysis). RESULTS: Data on hypoglycaemia and VD within the last 12 months were available for 3741 patients (98.2%) with a median (IQR) age of 65.9 (57.6-72.9) years; 46.7% were female. VD patients (n = 909; 24.3%) were older (70.7 vs 63.9 years; p < 0.0001), less often female (33.6% vs 50.9%; p < 0.0001) and had had diabetes for a longer duration (6.4 vs 5.4 years; p < 0.0001). Mean cholesterol (total, HDL and LDL) was also slightly lower (p < 0.0001). Glycaemic control (HbA1c, fasting and postprandial glucose) was comparable. VD patients received less metformin (80.7 vs 85.2%; p < 0.01) and more sulfonylureas (31.8 vs 27.6%; p < 0.05). There was an increased incidence of symptomatic hypoglycaemia with or without requiring help and with a need for medical assistance. After adjusting a number of baseline variables the rates of symptomatic hypoglycaemias with help remained significantly increased (OR 3.73 (95% CI 1.31-10.65) in patients with VD. CONCLUSIONS: As hypothesized there is a strong association between the incidence of hypoglycaemia and vascular disease at comparable glycaemic control, which confirms prior randomized controlled trial data suggesting an interrelationship between hypoglycaemia and vascular disease. PMID- 21628354 TI - The association of hypertension with renin-angiotensin system gene polymorphisms in the Lebanese population. AB - AIM: The study objective was to examine the association of hypertension in the Lebanese population with three renin-angiotensin system gene polymorphisms (RAS): angiotensin-converting enzyme (ACE), angiotensinogen (AGT) and angiotensin receptor type 1 (AT1R). METHODS: A total of 270 subjects (124 hypertensive vs 146 normotensive) were genotyped for ACE insertion (I)/deletion (D), AGT (M235T), and AT(1)R (A1166C) gene polymorphisms by polymerase chain reaction and restriction fragment length polymorphism. RESULTS: The studied genes showed no deviation from Hardy-Weinberg equilibrium. No association could be reported with the ACE I/D polymorphism, although the D allele frequency was high (77%) in patients. AGT TT genotype prevalence was found to be lower in hypertensive versus normotensive subjects (p<0.0001). AT(1)R CC and AC genotypes were significantly more frequent in hypertensive than normotensive subjects (p<0.0001). CONCLUSION: The first conducted study on the RAS gene polymorphisms in Lebanese hypertensive patients demonstrated a possible association of the AGT T and AT(1)R C alleles with hypertension. PMID- 21628355 TI - Long-term exercise attenuates blood pressure responsiveness and modulates kidney angiotensin II signalling and urinary sodium excretion in SHR. AB - Observations have been made regarding the effects of long-term exercise training on blood pressure, renal sodium handling and renal renin-angiotensin-aldosterone (RAS) intracellular pathways in conscious, trained Okamoto-Aoki spontaneously hypertensive rats (SHR) and Wistar Kyoto (WKy) normotensive rats, compared with appropriate age-matched sedentary SHR and WKy. To evaluate the influence of exercise training on renal function and RAS, receptors and intracellular angiotensin II (AngII) pathway compounds were used respectively, and lithium clearance and western blot methods were utilised. The current study demonstrated that increased blood pressure in SHR was blunted and significantly reduced by long-term swim training between the ages of 6 and 16 weeks. Additionally, the investigators observed an increased fractional urinary sodium excretion in trained SHR (SHR(T)) rats, compared with sedentary SHR (SHR(S)), despite a significantly decreased creatinine clearance (C(Cr)). Furthermore, immunoblotting analysis demonstrated a decreased expression of AT1(R) in the entire kidney of T(SHR) rats, compared with S(SHR). Conversely, the expression of the AT2(R), in both sedentary and trained SHR, was unchanged. The present study may indicate that, in the kidney, long-term exercise exerts a modulating effect on AngII receptor expression. In fact, the present study indicates an association of increasing natriuresis, reciprocal changes in renal AngII receptors and intracellular pathway proteins with the fall in blood pressure levels observed in T(SHR) rats compared with age-matched S(SHR) rats. PMID- 21628356 TI - Aliskiren reduces prorenin receptor expression and activity in cultured human aortic smooth muscle cells. AB - INTRODUCTION: The recent discovery of a specific receptor for renin/prorenin (PRR) has added new interest to the potential pharmacological actions of aliskiren, the first direct renin inhibitor. MATERIALS AND METHODS: In the present study, to gain new insights into the pharmacological properties of aliskiren, we investigated the effect of aliskiren on PRR expression and activity in cultured human smooth muscle cells (HSMCs). RESULTS: Co-incubation of HSMCs with angiotensinogen (ANG) (1.5 * 10(-7)M) and prorenin (10(-8)-10(-7)M) resulted in an efficient production (within 4h) of angiotensin I, almost completely inhibited by 10(-5)M aliskiren (-86.0 +/- 14.0%). In HSMCs stimulated with both ANG and prorenin, a 24h incubation with aliskiren (10(-6)-10(-5)M) resulted in a concentration-dependent reduction of PRR mRNA levels (IC(50) 4.6 * 10(-6)M). The cell surface expression of PRR determined by flow cytometry analysis was also reduced after incubation with aliskiren in a concentration-dependent manner. The lower levels of PRR were associated with a reduced expression of TGF-beta, PAI-1 and type I collagen mRNA. CONCLUSIONS: These results suggest a direct pharmacological action of aliskiren on PRR expression and its signalling pathway in HSMCs. This reported action of aliskiren may reveal a new scenario of the pharmacological properties of aliskiren. PMID- 21628357 TI - Factors associated with higher risks of emergency compulsory admission for immigrants: a report from the ASAP study. AB - OBJECTIVE: Specific ethnic groups appear to be disproportionally represented in emergency compulsory admissions. This may be due to a parallel higher risk of psychopathology, but different pathways to care in patients from ethnic minorities may also be an explanatory factor. In this article we concentrate on the influence of ethnic background, pathways to psychiatric emergency services and the amount of past psychiatric treatment as predictors of emergency compulsory admission. METHODS: All the patients coming into contact consecutively with the Psychiatric Emergency Service Amsterdam (PESA) from September 2004 to September 2006 were included in the study. We collected socio-demographic and clinical characteristics, and data about prior use of mental health services and referral to the emergency service. RESULTS: The risk of compulsory admission for immigrants from Surinam and the Dutch Antilles (OR 2.6), sub-Saharan Africa (OR 3.1), Morocco and other non-Western countries (each OR 1.7) was significantly higher than for native Dutch people. After controlling for socio-demographic characteristics, diagnosis, referral pattern and psychiatric treatment history, this correlation was no longer found. CONCLUSION: We found no evidence to substantiate the hypothesis that ethnic background plays an independent role in emergency compulsory admission. Police referral rather than referral by a GP, and being diagnosed with a psychosis, seemed to be explanatory factors for the high risk of compulsory admission for non-Western immigrants. Infrequent contact with secondary mental healthcare during the previous five years was not found to be consistently associated with a higher risk. PMID- 21628358 TI - Course and outcome of psychosis in black Caribbean populations and other ethnic groups living in the UK: a systematic review. AB - BACKGROUND: A higher incidence of psychosis has repeatedly been reported in black Caribbean populations in the UK. This has been attributed to a number of biological, psychological and sociocultural causes, including black Caribbean populations having a different illness course and outcome compared to other ethnic populations living in the UK. AIMS: A systematic review of UK-based quantitative studies, which compared at least two aspects of outcome in black Caribbean populations and other ethnic populations living in the UK, was conducted to assess whether the current body of research suggests that there are differences in the course and outcome of psychoses for these populations. METHOD: A wide variety of databases were searched using MeSH terms and keywords. Studies were evaluated according to specified inclusion criteria and analysed using predefined scoring criteria. RESULTS: Searches yielded a heterogeneous collection of studies. Large variances in methodological approaches and the quality of studies were reported. Many studies reported little or no difference between black Caribbean and other ethnic populations living in the UK. CONCLUSIONS: Emphasis is placed on the unreliability of these findings given the methodological limitations of the studies, and the need for higher-quality research in this area is highlighted. PMID- 21628359 TI - Differential item functioning due to gender between depression and anxiety items among Chilean adolescents. AB - BACKGROUND: Although much is known about the higher prevalence of anxiety and depressive disorders among adolescent females, less is known about the differential item endorsement due to gender in items of scales commonly used to measure anxiety and depression. AIMS: We conducted a study to examine if adolescent males and females from Chile differed on how they endorsed the items of the Youth Self Report (YSR) anxious/depressed problem scale. We used data from a cross-sectional sample consisting of 925 participants (mean age = 14, SD 1.3, 49% females) of low to lower-middle socioeconomic status. METHODS: A two parameter logistic (2PL) IRT DIF model was fit. RESULTS: RESULT: s revealed differential item functioning (DIF) by gender for six of the 13 items, with adolescent females being more likely to endorse a depression item while males were found more likely to endorse anxiety items. CONCLUSIONS: Findings suggest that items found in commonly used measures of anxiety and depression symptoms may not equally capture the true levels of these behavioural problems in adolescent males and females. Given the high levels of mental disorders in Chile and the surrounding countries, further attention should be focused on increasing the number of empirical studies examining potential gender differences in the assessment of mental health problems among Latin American populations to better aid our understanding of the phenomenology and determinants of these problems in the region. PMID- 21628360 TI - Vitamin D and cardiovascular disease: a jack of all trades? PMID- 21628362 TI - Vitamin D review. PMID- 21628363 TI - How we took control and became more efficient. PMID- 21628364 TI - Effect of weekly vitamin D supplements on mortality, morbidity, and growth of low birthweight term infants in India up to age 6 months: randomised controlled trial. AB - OBJECTIVE: To investigate whether vitamin D supplementation can decrease the mortality and morbidity of low birthweight infants in low income countries. DESIGN: Randomised controlled trial. SETTING: Large government hospital in New Delhi, India. PARTICIPANTS: 2079 low birthweight infants born at term (>37 weeks' gestation). MAIN OUTCOME MEASURES: Primary outcome was admission to hospital or death during the first six months of life. Main secondary outcome was growth. INTERVENTIONS: Weekly vitamin D supplements for six months at a dose of one recommended nutrient intake per day (35 ug/week). Infants were visited weekly at home for observed supplementation and were brought to the clinic monthly for clinical examination and anthropometric measurements. RESULTS: Between group differences were not significant for death or hospital admissions (92 among 1039 infants in the vitamin D group v 99 among 1040 infants in the placebo group; adjusted rate ratio 0.93, 95% confidence interval 0.68 to 1.29; P = 0.68), or referral to the outpatient clinic for moderate morbidity. Vitamin D supplementation resulted in better vitamin D status as assessed by plasma calcidiol levels at six months. In adjusted analyses, vitamin D treatment significantly increased standard deviation (z) scores at six months for weight, length, and arm circumference and decreased the proportion of children with stunted growth (length for age z score <= 2) or with arm circumference z scores of 2 or less. CONCLUSION: A weekly dose of vitamin D resulted in better vitamin D status and benefited the classic vitamin D function of bone growth but did not decrease the incidence of severe morbidity or death among young low birthweight infants. Trial registration ClinicalTrials.gov NCT00415402. PMID- 21628365 TI - Could routine cardiotocography reduce long term cognitive impairment? PMID- 21628366 TI - Time for an independent study. PMID- 21628367 TI - Venous thromboembolism is egregiously underestimated. PMID- 21628368 TI - Flawed meta-analysis creates doubt when answers are known. PMID- 21628369 TI - Study subject to unmeasured confounders and biases. PMID- 21628371 TI - Isotretinoin as contraindication. PMID- 21628372 TI - Jack Winkler replies to Joachim von Braun. PMID- 21628373 TI - Objective panels? Passion always trumps evidence. PMID- 21628374 TI - Diseases: on a continuum rather than discrete entities. PMID- 21628375 TI - Leadership in the NHS. PMID- 21628376 TI - WHO processes on dealing with a pandemic need to be overhauled and made more transparent. PMID- 21628377 TI - Trial of niacin alongside statin is stopped early. PMID- 21628378 TI - PCTs should live beyond 2013 to help GP consortiums bed in, says Nuffield trust. PMID- 21628379 TI - Outbreak of E coli in Germany is linked to cucumbers from Spain. PMID- 21628380 TI - Effective measures to prevent child injuries could help save 1000 lives a year. PMID- 21628381 TI - Major African clinical trial ends early due to high mortality in children. PMID- 21628382 TI - Government reverses policy on public health campaigns after numbers seeking advice fall sharply. PMID- 21628383 TI - Identification of novel TSH interaction sites by systematic binding analysis of the TSHR hinge region. AB - In which ways the binding of the thyroid stimulating hormone to the extracellular domain of its receptor leads to activation of the thyroid-stimulating hormone receptor (TSHR) is currently only incompletely understood. It is known that TSH binding to the TSHR depends on the interaction with the leucine-rich repeat and sulfation at Y385 of the hinge region. Recently it was also shown that electrostatic interactions between positive charges of bovine (b) TSH and the residues E297, E303, and D382 of the hinge region contribute to hormone-TSHR binding. After the identification of these first TSH binding sites in the hinge region, it was apparent that multiple positions in this region remained to be characterized for their roles in hormone binding. The goal of this study was therefore to clarify whether further contact points of TSH exist in the structurally undefined hinge region. Therefore, we systematically analyzed 41 uncharacterized residues of the TSHR hinge region as single mutants regarding differences between cell surface expression and bTSH binding. Indeed, we identified further amino acids of the hinge region with influence on bTSH binding. Some of these contribute to a new binding domain from human TSHR position F381 to D386. These hinge mutants with influence on bTSH binding were also analyzed for binding of the superagonistic human TSH analog TR1401 demonstrating that these positions also have an impact on TR1401 binding. Moreover, side chain variations revealed that different amino acid properties like the negative charge, aromatic as well as hydrophilic characteristics, contribute to maintain the hormone-TSHR hinge interaction. PMID- 21628384 TI - Type-2 iodothyronine 5'deiodinase in skeletal muscle of C57BL/6 mice. I. Identity, subcellular localization, and characterization. AB - RT-PCR shows that mouse skeletal muscle contains type-2 iodothyronine deiodinase (D2) mRNA. However, the D2 activity has been hard to measure. Except for newborn mice, muscle homogenates have no detectable activity. However, we have reported D2 activity in mouse muscle microsomes. As the mRNA, activity is higher in slow- than in fast-twitch muscle. We addressed here the major problems in measuring D2 activity in muscle by: homogenizing muscle in high salt to improve yield of membranous structures; separating postmitochondrial supernatant between 38 and 50% sucrose, to eliminate lighter membranes lacking D2; washing these with 0.1 M Na(2)CO(3) to eliminate additional contaminating proteins; pretreating all buffers with Chelex, to eliminate catalytic metals; and eliminating the EDTA from the assay, as this can bind iron that enhances dithiothreitol oxidation and promotes peroxidation reactions. Maximum velocity of T(3) generation by postgradient microsomes from red muscles was approximately 1100 fmol/(h . mg) protein with a Michaelis-Menten constant for T(4) of 1.5 nM. D2-specific activity of Na(2)CO(3)-washed microsomes was 6-10 times higher. The enrichment in D2 activity increased in parallel with the capacity of microsomes to load (sarco/endoplasmic reticulum Ca(2+)-ATPase) and bind Ca(2+) (calsequestrin), indicating that D2 resides in the inner sarcoplasmic reticulum, close to the nuclei. The presence of D3 in the sarcolemma suggests that the most of D2 generated T(3) acts locally. Estimates from maximum velocity, Michaelis-Menten constant, and muscle T(4) content suggest that mouse red, type-1, aerobic mouse muscle fibers can generate physiologically relevant amounts of T(3) and, further, that muscle D2 plays an important role in thyroid hormone-dependent muscle thermogenesis. PMID- 21628385 TI - STX, a novel nonsteroidal estrogenic compound, induces rapid action in primate GnRH neuronal calcium dynamics and peptide release. AB - Previously, we reported that 1 nM 17beta-estradiol (E(2)) induces a rapid action, which is, in part, mediated through the G protein-coupled receptor GPR30 in primate GnRH neurons. Because it has been reported that the diphenylacrylamide compound, STX, causes estrogenic action in the mouse and guinea pig hypothalamus, the present study examined effects of STX in primate GnRH neurons and whether there is an action independent of GPR30. Results are summarized as follows. STX (10 nM) exposure increased 1) the oscillation frequency of intracellular calcium concentration ([Ca(2+)](i)), 2) the percentage of cells stimulated, and 3) the synchronization frequency of [Ca(2+)](i) oscillations. STX (10-100 nM) also stimulated GnRH release. The effects of STX on both [Ca(2+)](i) oscillations and GnRH release were similar to those caused by E(2) (1 nM), although with less magnitude. STX (10 nM)-induced changes in [Ca(2+)](i) oscillations were not altered by GPR30 small interfering RNA transfection, indicating that STX sensitive receptors differ from GPR30. Finally, a higher dose of E(2) (10 nM) induced a larger change in [Ca(2+)](i) oscillations than that with a smaller dose of E(2) (1 nM), and the effects of 10 nM E(2) were reduced but not completely blocked by GPR30 small interfering RNA transfection, indicating that the effects of 10 nM E(2) in primate GnRH neurons are mediated by multiple membrane receptors, including GPR30 and STX-sensitive receptors. Collectively, the rapid action of E(2) mediated through GPR30 differs from that mediated through STX sensitive receptors. The molecular structure of the STX-sensitive receptor remains to be identified. PMID- 21628386 TI - Decreased IGF type 1 receptor signaling in mammary epithelium during pregnancy leads to reduced proliferation, alveolar differentiation, and expression of insulin receptor substrate (IRS)-1 and IRS-2. AB - The IGFs and the IGF type 1 receptor (IGF-1R) are essential mediators of normal mammary gland development in mice. IGF-I and the IGF-1R have demonstrated functions in formation and proliferation of terminal end buds and in ductal outgrowth and branching during puberty. To study the functions of IGF-1R during pregnancy and lactation, we established transgenic mouse lines expressing a human dominant-negative kinase dead IGF-1R (dnhIGF-1R) under the control of the whey acidic protein promoter. We provide evidence that the IGF-1R pathway is necessary for normal epithelial proliferation and alveolar formation during pregnancy. Furthermore, we demonstrate that the whey acidic protein-dnhIGF-1R transgene causes a delay in alveolar differentiation including lipid droplet formation, lumen expansion, and beta-casein protein expression. Analysis of IGF-1R signaling pathways showed a decrease in P-IGF-1R and P-Akt resulting from expression of the dnhIGF-1R. We further demonstrate that disruption of the IGF-1R decreases mammary epithelial cell expression of the signaling intermediates insulin receptor substrate (IRS)-1 and IRS-2. No alterations were observed in downstream signaling targets of prolactin and progesterone, suggesting that activation of the IGF-1R may directly regulate expression of IRS-1/2 during alveolar development and differentiation. These data show that IGF-1R signaling is necessary for normal alveolar proliferation and differentiation, in part, through induction of signaling intermediates that mediate alveolar development. PMID- 21628387 TI - Introduction of solids before 4 months is associated with obesity at 3 years among formula-fed infants but not among breast-fed infants. PMID- 21628388 TI - Effect of honey supplementation on the phagocytic function during nutritional rehabilitation of protein energy malnutrition patients. PMID- 21628389 TI - Residential aged care in Auckland, New Zealand 1988-2008: do real trends over time match predictions? AB - BACKGROUND: in Auckland, New Zealand in 1988, 7.7% of those aged over 65 years lived in licenced residential aged care. Age-specific rates approximately doubled for each 5-year age group after the age of 65 years. Even with changes in policies and market forces since 1988, population increases are forecast to drive large growth in demand. This study shows previously unrecognised 20-year trends in rates of care in a geographically defined population. METHODS: four cross sectional surveys of all facilities (rest homes and hospitals) licenced for long term care of older people were conducted in Auckland, New Zealand in 1988, 1993, 1998 and 2008. Facility staff completed survey forms for each resident. Numbers of licenced and occupied beds and trends in age-specific and age-standardised rates in residential aged care are reported. RESULTS: over the 20-year period, Auckland's population aged over 65 years increased by 43% (from 91,000 to 130,000) but actual numbers in care reduced slightly. Among those aged over 65 years, the proportion living in care facilities reduced from 1 in 13 to 1 in 18. Age-standardised rates in rest-home level care reduced from 65 to 33 per thousand, and in hospital level care, from 29 to 23 per thousand. Had rates remained stable, over 13,200 people, 74% more than observed, would have been in care in 2008. CONCLUSION: growth predicted in the residential aged care sector is not yet evident. The introduction of standardised needs assessments before entry, increased availability of home-based services, and growth in retirement villages may have led to reduced utilisation. PMID- 21628390 TI - Gait, gait variability and the risk of multiple incident falls in older people: a population-based study. AB - BACKGROUND: it is uncertain as to which measures of gait best predict those who are likely to fall. Our aim was to investigate the associations of gait and gait variability measures with incident falls risk. METHODS: individuals aged 60-86 years (n = 412) were randomly selected from the Tasmanian electoral roll. Average gait and gait variability measures were collected on a computerised walkway. Falls were recorded prospectively over 12 months. Log multinomial regression was used to estimate the relative risk of single and multiple falls associated with gait measures. Covariates included age, sex, sensorimotor and cognitive measures, mood and medications. RESULTS: in this population-based study greater intra individual variability in step length and double-support phase were linearly associated with increased risk of multiple falls (P = 0.04). Non-linear associations with multiple falls were found for gait speed P = 0.002, cadence P = 0.004 and step time variability P = 0.03. None of the gait measures predicted risk of single falls. CONCLUSION: there is an increased risk of multiple falls, but not single falls, in older people with poorer gait. Specific measures of gait and gait variability seem to confer this risk and may be amenable to interventions designed to reduce the risk of multiple falls in older people. PMID- 21628392 TI - Recombinant origin of the retrovirus XMRV. AB - The retrovirus XMRV (xenotropic murine leukemia virus-related virus) has been detected in human prostate tumors and in blood samples from patients with chronic fatigue syndrome, but these findings have not been replicated. We hypothesized that an understanding of when and how XMRV first arose might help explain the discrepant results. We studied human prostate cancer cell lines CWR22Rv1 and CWR R1, which produce XMRV virtually identical to the viruses recently found in patient samples, as well as their progenitor human prostate tumor xenograft (CWR22) that had been passaged in mice. We detected XMRV infection in the two cell lines and in the later passage xenografts, but not in the early passages. In particular, we found that the host mice contained two proviruses, PreXMRV-1 and PreXMRV-2, which share 99.92% identity with XMRV over >3.2-kilobase stretches of their genomes. We conclude that XMRV was not present in the original CWR22 tumor but was generated by recombination of two proviruses during tumor passaging in mice. The probability that an identical recombinant was generated independently is negligible (~10(-12)); our results suggest that the association of XMRV with human disease is due to contamination of human samples with virus originating from this recombination event. PMID- 21628393 TI - No evidence of murine-like gammaretroviruses in CFS patients previously identified as XMRV-infected. AB - Members of the gammaretroviruses--such as murine leukemia viruses (MLVs), most notably XMRV [xenotropic murine leukemia virus (X-MLV)-related virus--have been reported to be present in the blood of patients with chronic fatigue syndrome (CFS). We evaluated blood samples from 61 patients with CFS from a single clinical practice, 43 of whom had previously been identified as XMRV-positive. Our analysis included polymerase chain reaction and reverse transcription polymerase chain reaction procedures for detection of viral nucleic acids and assays for detection of infectious virus and virus-specific antibodies. We found no evidence of XMRV or other MLVs in these blood samples. In addition, we found that these gammaretroviruses were strongly (X-MLV) or partially (XMRV) susceptible to inactivation by sera from CFS patients and healthy controls, which suggested that establishment of a successful MLV infection in humans would be unlikely. Consistent with previous reports, we detected MLV sequences in commercial laboratory reagents. Our results indicate that previous evidence linking XMRV and MLVs to CFS is likely attributable to laboratory contamination. PMID- 21628394 TI - miRNA-223 promotes gastric cancer invasion and metastasis by targeting tumor suppressor EPB41L3. AB - Traditional research modes aim to find cancer-specific single therapeutic target. Recently, emerging evidence suggested that some micro-RNAs (miRNA) can function as oncogenes or tumor suppressors. miRNAs are single-stranded, small noncoding RNA genes that can regulate hundreds of downstream target genes. In this study, we evaluated the miRNA expression patterns in gastric carcinoma and the specific role of miR-223 in gastric cancer metastasis. miRNA expression signature was first analyzed by real-time PCR on 10 paired gastric carcinomas and confirmed in another 20 paired gastric carcinoma tissues. With the 2-fold expression difference as a cutoff level, we identified 22 differential expressed mature miRNAs. Sixteen miRNAs were upregulated in gastric carcinoma, including miR-223, miR-21, miR-23b, miR-222, miR-25, miR-23a, miR-221, miR-107, miR-103, miR-99a, miR-100, miR-125b, miR-92, miR-146a, miR-214 and miR-191, and six miRNAs were downregulated in gastric carcinoma, including let-7a, miR-126, miR-210, miR-181b, miR-197, and miR-30aa-5p. After examining these miRNAs in several human gastric originated cell lines, we found that miR-223 is overexpressed only in metastatic gastric cancer cells and stimulated nonmetastatic gastric cancer cells migration and invasion. Mechanistically, miR-223, induced by the transcription factor Twist, posttranscriptionally downregulates EPB41L3 expression by directly targeting its 3'-untranslated regions. Significantly, overexpression of miR-223 in primary gastric carcinomas is associated with poor metastasis-free survival. These findings indicate a new regulatory mode, namely, specific miRNA, which is activated by its upstream transcription factor, could suppress its direct targets and lead to tumor invasion and metastasis. PMID- 21628395 TI - Unbound (bioavailable) IGF1 enhances somatic growth. AB - Understanding insulin-like growth factor-1 (IGF1) biology is of particular importance because, apart from its role in mediating growth, it plays key roles in cellular transformation, organ regeneration, immune function, development of the musculoskeletal system and aging. IGF1 bioactivity is modulated by its binding to IGF-binding proteins (IGFBPs) and the acid labile subunit (ALS), which are present in serum and tissues. To determine whether IGF1 binding to IGFBPs is necessary to facilitate normal growth and development, we used a gene-targeting approach and generated two novel knock-in mouse models of mutated IGF1, in which the native Igf1 gene was replaced by Des-Igf1 (KID mice) or R3-Igf1 (KIR mice). The KID and KIR mutant proteins have reduced affinity for the IGFBPs, and therefore present as unbound IGF1, or 'free IGF1'. We found that both KID and KIR mice have reduced serum IGF1 levels and a concomitant increase in serum growth hormone levels. Ternary complex formation of IGF1 with the IGFBPs and the ALS was markedly reduced in sera from KID and KIR mice compared with wild type. Both mutant mice showed increased body weight, body and bone lengths, and relative lean mass. We found selective organomegaly of the spleen, kidneys and uterus, enhanced mammary gland complexity, and increased skeletal acquisition. The KID and KIR models show unequivocally that IGF1-complex formation with the IGFBPs is fundamental for establishing normal body and organ size, and that uncontrolled IGF bioactivity could lead to pathological conditions. PMID- 21628397 TI - The influence of optic expansion rates when judging the relative time to contact of familiar objects. AB - Previous research has shown that size-dependent errors in time-to-contact (TTC) judgments can be attenuated when approaching objects are familiar to the observer and have a known size. We describe two experiments that show that the effect of size on relative TTC judgments can be modeled on observers' reliance on the instantaneous optic expansion rates of the approaching objects. This reliance on optic expansion rates occurred independently of object familiarity and when the actual TTC of the approaching objects was relatively brief or relatively long. However, observers' sensitivity to differences in TTC was improved for familiar objects when TTC was large. These results are consistent with other research showing that optic expansion rate is a critical variable for judging TTC. PMID- 21628396 TI - Hadp1, a newly identified pleckstrin homology domain protein, is required for cardiac contractility in zebrafish. AB - The vertebrate heart is one of the first organs to form, and its early function and morphogenesis are crucial for continued embryonic development. Here we analyze the effects of loss of Heart adaptor protein 1 (Hadp1), which we show is required for normal function and morphogenesis of the embryonic zebrafish heart. Hadp1 is a pleckstrin homology (PH)-domain-containing protein whose expression is enriched in embryonic cardiomyocytes. Knockdown of hadp1 in zebrafish embryos reduced cardiac contractility and altered late myocyte differentiation. By using optical mapping and submaximal levels of hadp1 knockdown, we observed profound effects on Ca(2+) handling and on action potential duration in the absence of morphological defects, suggesting that Hadp1 plays a major role in the regulation of intracellular Ca(2+) handling in the heart. Hadp1 interacts with phosphatidylinositol 4-phosphate [PI4P; also known as PtdIns(4)P] derivatives via its PH domain, and its subcellular localization is dependent upon this motif. Pharmacological blockade of the synthesis of PI4P derivatives in vivo phenocopied the loss of hadp1 in zebrafish. Collectively, these results demonstrate that hadp1 is required for normal cardiac function and morphogenesis during embryogenesis, and suggest that hadp1 modulates Ca(2+) handling in the heart through its interaction with phosphatidylinositols. PMID- 21628398 TI - Identification and functional characterization in vivo of a novel splice variant of LDLR in rhesus macaques. AB - In the course of developing a low-density lipoprotein receptor (LDLR) gene therapy treatment for homozygous familial hypercholesterolemia (HoFH), we planned to examine the efficacy in a nonhuman primate model, the rhesus macaque heterozygous for an LDL receptor mutation fed a high-fat diet. Unexpectedly, our initial cDNA sequencing studies led to the identification of a heretofore unidentified splicing isoform of the rhesus LDLR gene. Compared with the publicly available GenBank reference sequence of rhesus LDLR, the novel isoform contains a 21 bp in frame insertion. This sequence coincides with part of exon 5 and creates a site for the restriction enzyme MscI. Using this site as a marker for the 21 bp in-frame insertion, we conducted a restriction enzyme screen to examine for the prevalence of the novel isoform in rhesus liver tissue cDNA and its homolog in human liver tissue cDNA. We found that the novel isoform is the predominant LDLR cDNA found in rhesus liver and the sole LDLR cDNA found in human liver. Finally, we compared the in vivo functionality of the novel and previously identified rhesus LDLR splicing isoforms in a mouse model of HoFH. PMID- 21628399 TI - Iron chelation with deferasirox in adult and pediatric patients with thalassemia major: efficacy and safety during 5 years' follow-up. AB - Patients with beta-thalassemia require lifelong iron chelation therapy from early childhood to prevent complications associated with transfusional iron overload. To evaluate long-term efficacy and safety of once-daily oral iron chelation with deferasirox, patients aged >= 2 years who completed a 1-year, phase 3, randomized trial entered a 4-year extension study, either continuing on deferasirox (deferasirox cohort) or switching from deferoxamine to deferasirox (crossover cohort). Of 555 patients who received >= 1 deferasirox dose, 66.8% completed the study; 43 patients (7.7%) discontinued because of adverse events. In patients with >= 4 years' deferasirox exposure who had liver biopsy, mean liver iron concentration significantly decreased by 7.8 +/- 11.2 mg Fe/g dry weight (dw; n = 103; P < .001) and 3.1 +/- 7.9 mg Fe/g dw (n = 68; P < .001) in the deferasirox and crossover cohorts, respectively. Median serum ferritin significantly decreased by 706 ng/mL (n = 196; P < .001) and 371 ng/mL (n = 147; P < .001), respectively, after >= 4 years' exposure. Investigator-assessed, drug-related adverse events, including increased blood creatinine (11.2%), abdominal pain (9.0%), and nausea (7.4%), were generally mild to moderate, transient, and reduced in frequency over time. No adverse effect was observed on pediatric growth or adolescent sexual development. This first prospective study of long term deferasirox use in pediatric and adult patients with beta-thalassemia suggests treatment for <= 5 years is generally well tolerated and effectively reduces iron burden. This trial was registered at www.clinicaltrials.gov as #NCT00171210. PMID- 21628400 TI - Allogeneic hematopoietic stem cell transplantation for sickle cell disease: the time is now. AB - Although sickle cell disease (SCD) has a variable clinical course, many patients develop end-organ complications that are associated with significant morbidity and early mortality. Myeloablative allogeneic HSCT (allo-HSCT) is curative but has been historically performed only in children younger than 16 years of age. Modest modifications in the conditioning regimen and supportive care have improved outcome such that the majority of children with a suitable HLA-matched sibling donor can expect a cure from this approach. However, adult patients have been excluded from myeloablative allo-HSCT because of anticipated excess toxicity resulting from accumulated disease burden. Efforts to use nonmyeloablative transplantation strategies in adults logically followed but were initially met with largely disappointing results. Recent results, however, indicate that nonmyeloablative allo-HSCT in adult patients with SCD allows for stable mixed hematopoietic chimerism with associated full-donor erythroid engraftment and normalization of blood counts, and persistence in some without continued immunosuppression suggests immunologic tolerance. The attainment of tolerance should allow extension of these potentially curative approaches to alternative donor sources. Efforts to build on these experiences should increase the use of allo-HSCT in patients with SCD while minimizing morbidity and mortality. PMID- 21628401 TI - Megakaryocytes differentially sort mRNAs for matrix metalloproteinases and their inhibitors into platelets: a mechanism for regulating synthetic events. AB - Megakaryocytes transfer a diverse and functional transcriptome to platelets during the final stages of thrombopoiesis. In platelets, these transcripts reflect the expression of their corresponding proteins and, in some cases, serve as a template for translation. It is not known, however, if megakaryocytes differentially sort mRNAs into platelets. Given their critical role in vascular remodeling and inflammation, we determined whether megakaryocytes selectively dispense transcripts for matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) into platelets. Next-generation sequencing (RNA-Seq) revealed that megakaryocytes express mRNA for 10 of the 24 human MMP family members. mRNA for all of these MMPs are present in platelets with the exception of MMP-2, 14, and 15. Megakaryocytes and platelets also express mRNA for TIMPs 1-3, but not TIMP-4. mRNA expression patterns predicted the presence and, in most cases, the abundance of each corresponding protein. Nonetheless, exceptions were observed: MMP-2 protein is present in platelets but not its transcript. In contrast, quiescent platelets express TIMP-2 mRNA but only traces of TIMP-2 protein. In response to activating signals, however, platelets synthesize significant amounts of TIMP-2 protein. These results demonstrate that megakaryocytes differentially express mRNAs for MMPs and TIMPs and selectively transfer a subset of these into platelets. Among the platelet messages, TIMP-2 serves as a template for signal dependent translation. PMID- 21628402 TI - The novel anti-MEK small molecule AZD6244 induces BIM-dependent and AKT independent apoptosis in diffuse large B-cell lymphoma. AB - The RAS/RAF/MEK/ERK signaling pathway has been largely unexplored as a potential therapeutic target in lymphoma. The novel 2nd generation anti-MEK small molecule, AZD6244, down-regulated its direct downstream target, phospho-ERK (pERK) in germinal center and nongerminal center diffuse large B-cell lymphoma (DLBCL) cell lines and primary cells. Similar decreased pERK levels were noted despite constitutive activation (CA) of MEK. Consequently, several lymphoma-related ERK substrates were down-regulated by AZD6244 including MCT-1, c-Myc, Bcl-2, Mcl-1, and CDK1/2. AZD6244 induced time- and dose-dependent antiproliferation and apoptosis in all DLBCL cell lines and fresh/primary cells (IC(50) 100nM-300nM). Furthermore, AZD6244 resulted in significantly less tumor compared with control in an in vivo DLBCL SCID xenograft model. Cell death was associated with cleaved PARP, caspases-8, -9, and -3, and apoptosis was caspase-dependent. In addition, there was stabilization of FoxO3a, activation of BIM and PUMA, and a significant decrease in c-Myc transcripts. Moreover, siRNA knockdown of BIM abrogated AZD6244 related apoptosis, while shRNA knockdown of ERK minimally sensitized cells. Finally, manipulation of AKT with transfection of OCI-LY3 cells with CA-AKT or through chemical inhibition (LY294002) had minimal effect on AZD6244-induced cell death. Altogether, these findings show that the novel anti-MEK agent, AZD6244, induced apoptosis in DLBCL and that cell death was BIM-dependent. PMID- 21628403 TI - Modeling the evolution of ETV6-RUNX1-induced B-cell precursor acute lymphoblastic leukemia in mice. AB - The t(12;21) translocation that generates the ETV6-RUNX1 (TEL-AML1) fusion gene, is the most common chromosomal rearrangement in childhood cancer and is exclusively associated with B-cell precursor acute lymphoblastic leukemia (BCP ALL). The translocation arises in utero and is necessary but insufficient for the development of leukemia. Single-nucleotide polymorphism array analysis of ETV6 RUNX1 patient samples has identified multiple additional genetic alterations; however, the role of these lesions in leukemogenesis remains undetermined. Moreover, murine models of ETV6-RUNX1 ALL that faithfully recapitulate the human disease are lacking. To identify novel genes that cooperate with ETV6-RUNX1 in leukemogenesis, we generated a mouse model that uses the endogenous Etv6 locus to coexpress the Etv6-RUNX1 fusion and Sleeping Beauty transposase. An insertional mutagenesis screen was performed by intercrossing these mice with those carrying a Sleeping Beauty transposon array. In contrast to previous models, a substantial proportion (20%) of the offspring developed BCP-ALL. Isolation of the transposon insertion sites identified genes known to be associated with BCP-ALL, including Ebf1 and Epor, in addition to other novel candidates. This is the first mouse model of ETV6-RUNX1 to develop BCP-ALL and provides important insight into the cooperating genetic alterations in ETV6-RUNX1 leukemia. PMID- 21628404 TI - Adam17-dependent shedding limits early neutrophil influx but does not alter early monocyte recruitment to inflammatory sites. AB - TNF-alpha-converting enzyme (TACE, herein denoted as Adam17) proteolytically sheds several cell-surface inflammatory proteins, but the physiologic importance of the cleavage of these substrates from leukocyte subsets during inflammation is incompletely understood. In this study, we show that Adam17-null neutrophils have a 2-fold advantage in their initial recruitment during thioglycollate-induced peritonitis, and they roll slower and adhere more readily in the cremaster model than wild-type neutrophils. Although CD44 and ICAM-1 are both in vitro substrates of Adam17, their surface levels are not altered on Adam17-null neutrophils. In contrast, L-selectin levels are elevated up to 10-fold in Adam17-null circulating neutrophils, and their accelerated peritoneal influx, slower rolling, and increased adhesion in the cremaster muscle are dependent on L-selectin. Analysis of mixed chimeras shows that enhanced L-selectin levels and accelerated influx were both cell-intrinsic properties of neutrophils lacking Adam17. In contrast, Adam17-null monocytes display no acceleration of infiltration into the peritoneum in spite of elevated L-selectin surface levels, and their peritoneal influx was independent of L-selectin. Therefore, our data demonstrate substrate and myeloid cell-type specificity of Adam17-mediated cleavage of its substrates, and show that neutrophils and monocytes use distinct mechanisms for infiltration of tissues. PMID- 21628405 TI - FLT3 expression initiates in fully multipotent mouse hematopoietic progenitor cells. AB - Lymphoid-primed multipotent progenitors with down-regulated megakaryocyte erythroid (MkE) potential are restricted to cells with high levels of cell surface FLT3 expression, whereas HSCs and MkE progenitors lack detectable cell surface FLT3. These findings are compatible with FLT3 cell-surface expression not being detectable in the fully multipotent stem/progenitor cell compartment in mice. If so, this process could be distinct from human hematopoiesis, in which FLT3 already is expressed in multipotent stem/progenitor cells. The expression pattern of Flt3 (mRNA) and FLT3 (protein) in multipotent progenitors is of considerable relevance for mouse models in which prognostically important Flt3 mutations are expressed under control of the endogenous mouse Flt3 promoter. Herein, we demonstrate that mouse Flt3 expression initiates in fully multipotent progenitors because in addition to lymphoid and granulocyte-monocyte progenitors, FLT3(-) Mk- and E-restricted downstream progenitors are also highly labeled when Flt3-Cre fate mapping is applied. PMID- 21628406 TI - Reprogramming of EBV-immortalized B-lymphocyte cell lines into induced pluripotent stem cells. AB - EBV-immortalized B lymphocyte cell lines have been widely banked for studying a variety of diseases, including rare genetic disorders. These cell lines represent an important resource for disease modeling with the induced pluripotent stem cell (iPSC) technology. Here we report the generation of iPSCs from EBV-immortalized B cell lines derived from multiple inherited disease patients via a nonviral method. The reprogramming method for the EBV cell lines involves a distinct protocol compared with that of patient fibroblasts. The B-cell line-derived iPSCs expressed pluripotency markers, retained the inherited mutation and the parental V(D)J rearrangement profile, and differentiated into all 3 germ layer cell types. There was no integration of the reprogramming-related transgenes or the EBV associated genes in these iPSCs. The ability to reprogram the widely banked patient B-cell lines will offer an unprecedented opportunity to generate human disease models and provide novel drug therapies. PMID- 21628407 TI - Chromothripsis identifies a rare and aggressive entity among newly diagnosed multiple myeloma patients. AB - Multiple myeloma (MM) develops from a premalignant plasma cell proliferative disorder, and with time can progress to a more aggressive disease in extramedullary locations. The gradually clinical evolution is supported by clonal expansion of cells that acquire genetic lesions over years. This model of cancer evolution based on ongoing genomic instability mechanism may apply to development of most MM cases. However, in a small fraction of newly diagnosed MM who relapse quickly and finally die within 2 years, the gradual model appears to be untenable. Analysis of high resolution copy number profiles obtained using single nucleotide polymorphism array data from 764 newly diagnosed MM identified large numbers of genomic rearrangements with the hallmarks of chromothripsis in 1.3% of samples. Moreover, this catastrophic event confers a poor outcome. Because chromothripsis appears to occur in a single crisis, our results suggest that high risk MM patients use this novel way of cancer evolution. PMID- 21628408 TI - Pharmacogenomics of bortezomib test-dosing identifies hyperexpression of proteasome genes, especially PSMD4, as novel high-risk feature in myeloma treated with Total Therapy 3. AB - Gene expression profiling (GEP) of purified plasma cells 48 hours after thalidomide and dexamethasone test doses showed these agents' mechanisms of action and provided prognostic information for untreated myeloma patients on Total Therapy 2 (TT2). Bortezomib was added in Total Therapy 3 (TT3), and 48 hours after bortezomib GEP analysis identified 80 highly survival-discriminatory genes in a training set of 142 TT3A patients that were validated in 128 patients receiving TT3B. The 80-gene GEP model (GEP80) also distinguished outcomes when applied at baseline in both TT3 and TT2 protocols. In context of our validated 70 gene model (GEP70), the GEP80 model identified 9% of patients with a grave prognosis among those with GEP70-defined low-risk disease and 41% of patients with favorable prognosis among those with GEP70-defined high-risk disease. PMSD4 was 1 of 3 genes common to both models. Residing on chromosome 1q21, PSMD4 expression is highly sensitive to copy number. Both higher PSMD4 expression levels and higher 1q21 copy numbers affected clinical outcome adversely. GEP80 baseline-defined high risk, high lactate dehydrogenase, and low albumin were the only independent adverse variables surviving multivariate survival model. We are investigating whether second-generation proteasome inhibitors (eg, carfilzomib) can overcome resistance associated with high PSMD4 levels. PMID- 21628409 TI - RhoJ is an endothelial cell-restricted Rho GTPase that mediates vascular morphogenesis and is regulated by the transcription factor ERG. AB - ERG is a member of the ETS transcription factor family that is highly enriched in endothelial cells (ECs). To further define the role of ERG in regulating EC function, we evaluated the effect of ERG knock-down on EC lumen formation in 3D collagen matrices. Blockade of ERG using siRNA completely interferes with EC lumen formation. Quantitative PCR (QPCR) was used to identify potential downstream gene targets of ERG. In particular, we identified RhoJ as the Rho GTPase family member that is closely related to Cdc42 as a target of ERG. Knockdown of ERG expression in ECs led to a 75% reduction in the expression of RhoJ. Chromatin immunoprecipitation and transactivation studies demonstrated that ERG could bind to functional sites in the proximal promoter of the RhoJ gene. Knock-down of RhoJ similarly resulted in a marked reduction in the ability of ECs to form lumens. Suppression of either ERG or RhoJ during EC lumen formation was associated with a marked increase in RhoA activation and a decrease in Rac1 and Cdc42 activation and their downstream effectors. Finally, in contrast to other Rho GTPases, RhoJ exhibits a highly EC-restricted expression pattern in several different tissues, including the brain, heart, lung, and liver. PMID- 21628410 TI - Neurexophilin 1 suppresses the proliferation of hematopoietic progenitor cells. AB - Neurexin I alpha (NRXN1alpha) and Dystroglycan (DAG1) are membrane receptors which serve as mutual ligands in the neuronal system. Neurexophilins (NXPHs) bind NRXN1alpha. NRXN1alpha was expressed in primitive populations in human CB (huCB) and murine BM (muBM). DAG1 is ubiquitously expressed in hematopoietic tissue; however, osteoblasts appear to be sites of very high expression within muBM. High concentrations of NXPH were found in huCB plasma and murine lineage-positive splenocytes. We evaluated effects of these molecules on huCB and muBM hematopoietic progenitor cells (HPCs) and HSCs. At both a single and population cell level in vitro, we found that NXPH1 was a potent inhibitor of HPC proliferation acting through NRXN1alpha an effect down-modulated by DAG1. Injection of recombinant NXPH1 in vivo resulted in myelo- and lymphosuppression in the BM, with absolute numbers and cycling status of functional and phenotypically defined HPCs dose- and time-dependently decreased. Competitive HSC transplantations showed no change in the long-term repopulating activity of HSCs from mice exposed to recombinant NXPH1. These results demonstrate the presence and function of a regulated signaling axis in hematopoiesis centered on NRXN1alpha and its modulation by DAG1 and NXPH1. PMID- 21628411 TI - Mechanisms of cytotoxicity to Pim kinase inhibitor, SGI-1776, in acute myeloid leukemia. AB - Pim kinases are Ser/Thr kinases with multiple substrates that affect survival pathways. These proteins are overexpressed in acute myeloid leukemia (AML) blasts and we hypothesized that Pim kinase inhibition would affect AML cell survival. Imidazo[1,2-b]pyridazine compound, SGI-1776 inhibits Pim-1, Pim-2 and Pim-3, and was evaluated in AML-cell line, -xenograft model, and -primary blasts. Treatment of AML cells with SGI-1776 results in a concentration-dependent induction of apoptosis and we investigated its effect on Pim kinase functions. Phosphorylation of traditional Pim kinase targets, c-Myc(Ser62) and 4E-BP1 (Thr36/Thr47), were both decreased in actively cycling AML cell lines MV-4-11, MOLM-13 and OCI-AML-3. Levels of antiapoptotic proteins Bcl-2, Bcl-x(L), XIAP, and proapoptotic Bak and Bax were unchanged; however, a significant reduction in Mcl-1 was observed. This was correlated with inhibition of global RNA and protein synthesis and MCL-1 transcript decline after SGI-1776 treatment. These data suggest that SGI-1776 mechanism in AML involves Mcl-1 protein reduction. Consistent with cell line data, xenograft model studies with mice bearing MV-4-11 tumors showed efficacy with SGI-1776. Importantly, SGI-1776 was also cytotoxic in AML primary cells, irrespective of FLT3 mutation status and resulted in Mcl-1 protein decline. Pim kinase inhibition may be a new strategy for AML treatment. PMID- 21628412 TI - c-Maf plays a crucial role for the definitive erythropoiesis that accompanies erythroblastic island formation in the fetal liver. AB - c-Maf is one of the large Maf (musculoaponeurotic fibrosarcoma) transcription factors that belong to the activated protein-1 super family of basic leucine zipper proteins. Despite its overexpression in hematologic malignancies, the physiologic roles c-Maf plays in normal hematopoiesis have been largely unexplored. On a C57BL/6J background, c-Maf(-/-) embryos succumbed from severe erythropenia between embryonic day (E) 15 and E18. Flow cytometric analysis of fetal liver cells showed that the mature erythroid compartments were significantly reduced in c-Maf(-/-) embryos compared with c-Maf(+/+) littermates. Interestingly, the CFU assay indicated there was no significant difference between c-Maf(+/+) and c-Maf(-/-) fetal liver cells in erythroid colony counts. This result indicated that impaired definitive erythropoiesis in c-Maf(-/-) embryos is because of a non-cell-autonomous effect, suggesting a defective erythropoietic microenvironment in the fetal liver. As expected, the number of erythroblasts surrounding the macrophages in erythroblastic islands was significantly reduced in c-Maf(-/-) embryos. Moreover, decreased expression of VCAM-1 was observed in c-Maf(-/-) fetal liver macrophages. In conclusion, these results strongly suggest that c-Maf is crucial for definitive erythropoiesis in fetal liver, playing an important role in macrophages that constitute erythroblastic islands. PMID- 21628413 TI - Low hepcidin accounts for the proinflammatory status associated with iron deficiency. AB - Hepcidin is an antimicrobial peptide that controls systemic iron homeostasis. Hepcidin binding to its receptor ferroportin reduces iron availability, thus controlling microbial growth. In parallel it triggers an anti-inflammatory response in macrophages. Hepcidin is transcriptionally regulated by iron, through the bone morphogenetic protein-son of mothers against decapentaplegic (BMP-SMAD) pathway and by inflammation, through IL6-mediated STAT3 signaling. To investigate the mechanisms linking iron and inflammation, we treated C57BL/6 iron-deficient mice with a sublethal dose of lipopolysaccharide (LPS) and analyzed their inflammatory response in comparison with controls. We show that iron-deprived mice have a proinflammatory condition, exacerbated by LPS treatment leading to increased IL6 and TNFalpha mRNA in liver and spleen macrophages, and increased serum IL6 (482.29 +/- 205.59 pg/mL) versus controls (69.01 +/- 17.52 pg/mL; P < .05). Hepcidin was undetectable in iron-deficient mice but pretreatment with hepcidin normalized their response to LPS. Tmprss6(-/-) mice, characterized by iron deficiency and high hepcidin, show a blunted inflammatory response when challenged with LPS. Our data support a model in which the lack of hepcidin is responsible of the high inflammatory response to LPS in iron deficiency. The proinflammatory status associated with chronic iron deficiency could explain the resistance to infection seen in this condition. PMID- 21628414 TI - Comprehensive analysis of mammalian miRNA* species and their role in myeloid cells. AB - Processing of pre-miRNA through Dicer1 generates an miRNA duplex that consists of an miRNA and miRNA* strand. Despite the general view that miRNA*s have no functional role, we further investigated miRNA* species in 10 deep-sequencing libraries from mouse and human tissue. Comparisons of miRNA/miRNA* ratios across the miRNA sequence libraries revealed that 50% of the investigated miRNA duplexes exhibited a highly dominant strand. Conversely, 10% of miRNA duplexes showed a comparable expression of both strands, whereas the remaining 40% exhibited variable ratios across the examined libraries, as exemplified by miR-223/miR-223* in murine and human cell lines. Functional analyses revealed a regulatory role for miR-223* in myeloid progenitor cells, which implies an active role for both arms of the miR-223 duplex. This was further underscored by the demonstration that miR-223 and miR-223* targeted the insulin-like growth factor 1 receptor/phosphatidylinositol 3-kinase axis and that high miR-223* levels were associated with increased overall survival in patients with acute myeloid leukemia. Thus, we found a supporting role for miR-223* in differentiating myeloid cells in normal and leukemic cell states. The fact that the miR-223 duplex acts through both arms extends the complexity of miRNA-directed gene regulation of this myeloid key miRNA. PMID- 21628415 TI - Long-term restoration of the human T-cell compartment after thymectomy during infancy: a role for thymic regeneration? AB - Thymectomy during early childhood is generally thought to have serious consequences for the establishment of the T-cell compartment. In the present study, we investigated the composition of the T-cell pool in the first 3 decades after thymectomy during infancy due to cardiac surgery. In the first 5 years after thymectomy, naive and total CD4(+) and CD8(+) T-cell numbers in the blood and T-cell receptor excision circle (TREC) levels in CD4(+) T cells were significantly lower than in healthy age-matched controls. In the first years after thymectomy, plasma IL-7 levels were significantly elevated and peripheral T cell proliferation levels were increased by ~ 2-fold. From 5 years after thymectomy onward, naive CD4(+) and CD8(+) T-cell counts and TRECs were within the normal range. Because TREC levels are expected to decline continuously in the absence of thymic output, we investigated whether normalization of the naive T cell pool could be due to regeneration of thymic tissue. In the majority of individuals who had been thymectomized during infancy, thymic tissue could indeed be identified on magnetic resonance imaging scans. Whereas thymectomy has severe effects on the establishment of the naive T-cell compartment during early childhood, our data suggest that functional regrowth of thymic tissue can limit its effects in subsequent years. PMID- 21628416 TI - Genetic variation in recipient B-cell activating factor modulates phenotype of GVHD. AB - B-cell activating factor (BAFF) single nucleotide polymorphisms (SNPs) are associated with autoimmune diseases. Because patients with classic and overlap chronic GVHD (cGVHD) have features of autoimmune diseases, we studied the association of recipient and/or donor BAFF SNPs with the phenotype of GVHD after allogeneic stem cell transplantation. Twenty tagSNPs of the BAFF gene were genotyped in 164 recipient/donor pairs. GVHD after day 100 occurred in 124 (76%) patients: acute GVHD (aGVHD) subtypes (n = 23), overlap GVHD (n = 29), and classic cGVHD (n = 72). In SNP analyses, 9 of the 20 tag SNPs were significant comparing classic/overlap cGVHD versus aGVHD subtypes/no GVHD. In multivariate analyses, 4 recipient BAFF SNPs (rs16972217 [odds ratio = 2.72, P = .004], rs7993590 [odds ratio = 2.35, P = .011], rs12428930 [odds ratio2.53, P = .008], and rs2893321 [odds ratio = 2.48, P = .009]) were independent predictors of GVHD subtypes, adjusted for conventional predictors of cGVHD. This study shows that genetic variation of BAFF modulates GVHD phenotype after allogeneic stem cell transplantation. PMID- 21628417 TI - MiR-17/20/93/106 promote hematopoietic cell expansion by targeting sequestosome 1 regulated pathways in mice. AB - MicroRNAs (miRNAs) are pivotal for regulation of hematopoiesis but their critical targets remain largely unknown. Here, we show that ectopic expression of miR-17, 20,-93 and -106, all AAAGUGC seed-containing miRNAs, increases proliferation, colony outgrowth and replating capacity of myeloid progenitors and results in enhanced P-ERK levels. We found that these miRNAs are endogenously and abundantly expressed in myeloid progenitors and down-regulated in mature neutrophils. Quantitative proteomics identified sequestosome 1 (SQSTM1), an ubiquitin-binding protein and regulator of autophagy-mediated protein degradation, as a major target for these miRNAs in myeloid progenitors. In addition, we found increased expression of Sqstm1 transcripts during CSF3-induced neutrophil differentiation of 32D-CSF3R cells and an inverse correlation of SQSTM1 protein levels and miR 106 expression in AML samples. ShRNA-mediated silencing of Sqstm1 phenocopied the effects of ectopic miR-17/20/93/106 expression in hematopoietic progenitors in vitro and in mice. Further, SQSTM1 binds to the ligand-activated colony stimulating factor 3 receptor (CSF3R) mainly in the late endosomal compartment, but not in LC3 positive autophagosomes. SQSTM1 regulates CSF3R stability and ligand-induced mitogen-activated protein kinase signaling. We demonstrate that AAAGUGC seed-containing miRNAs promote cell expansion, replating capacity and signaling in hematopoietic cells by interference with SQSTM1-regulated pathways. PMID- 21628418 TI - Aromatase promoter I.f is regulated by progesterone receptor in mouse hypothalamic neuronal cell lines. AB - Aromatase catalyzes the conversion of C(19) steroids to estrogens. Aromatase and progesterone, both of which function at different steps of steroidogenesis, are crucial for the sexually dimorphic development of the fetal brain and the regulation of gonadotropin secretion and sexual interest in adults. The aromatase gene (Cyp19a1) is selectively expressed in distinct neurons of the mouse hypothalamus through a distal brain-specific promoter, I.f, located ~40 kb upstream of the coding region. However, the regulation of aromatase expression in the brain is not well understood. In this study, we investigated a short feedback effect of progesterone analogues on aromatase mRNA expression and enzyme activity in estrogen receptor alpha (Esr1)-positive or -negative mouse embryonic hypothalamic neuronal cell lines that express aromatase via promoter I.f. In a hypothalamic neuronal cell line that highly expresses aromatase, progesterone receptor (Pgr), and Esr1, a progesterone agonist, R5020, inhibited aromatase mRNA level and enzyme activity. The inhibitory effect of R5020 was reversed by its antagonist, RU486. Deletion mutants of promoter I.f suggested that inhibition of aromatase expression by progesterone is conferred by the nt -1000/-500 region, and R5020 enhanced binding of Pgr to the nt -800/-600 region of promoter I.f. Small interfering RNA knockdown of Pgr eliminated progesterone-dependent inhibition of aromatase mRNA and enzyme activity. Taken together, progesterone enhances recruitment of Pgr to specific regions of the promoter I.f of Cyp19a1 and regulates aromatase expression in hypothalamic neurons. PMID- 21628419 TI - Retinoic acid isomers up-regulate ATP binding cassette A1 and G1 and cholesterol efflux in rat astrocytes: implications for their therapeutic and teratogenic effects. AB - Recent studies suggest that retinoids may be effective in the treatment of Alzheimer's disease, although exposure to an excess of retinoids during gestation causes teratogenesis. Cholesterol is essential for brain development, but high levels of cholesterol have been associated with Alzheimer's disease. We hypothesized that retinoic acid may affect cholesterol homeostasis in rat astrocytes, which regulate cholesterol distribution in the brain, through the up regulation of cholesterol transporters ATP binding cassette (Abc)a1 and Abcg1. Tretinoin, 13-cis retinoic acid (13-cis-RA), 9-cis-RA, and the selective retinoid X receptor (RXR) agonist methoprene significantly increased cholesterol efflux induced by cholesterol acceptors and protein levels of Abca1 by 2.3- (+/- 0.25), 3.6- (+/- 0.42), 4.1- (+/- 0.5), and 1.75- (+/- 0.43) fold, respectively, and Abcg1 by 2.1- (+/- 0.26), 2.2- (+/- 0.33), 2.5- (+/- 0.23), and 2.2- (+/- 0.21) fold, respectively. 13-cis-RA and 9-cis-RA also significantly increased mRNA levels of Abca1 (maximal induction 7.3 +/- 0.42 and 2.7 +/- 0.17, respectively) and Abcg1 (maximal induction 2.0 +/- 0.18 and 1.8 +/- 0.09, respectively), and the levels of membrane-bound Abca1 (2.5 +/- 0.3 and 2.5 +/- 0.40-fold increase, respectively), whereas they significantly decreased intracellular cholesterol content without affecting cholesterol synthesis. The effect of 9-cis-RA on cholesterol homeostasis in astrocytes can be ascribed to the activation of RXR, whereas the effects of 13-cis-RA and tretinoin were independent of either RXRs or retinoic acid receptors. These findings suggest that retinoids affect cholesterol homeostasis in astrocytes and that this effect may be involved in both their therapeutic and teratogenic actions. PMID- 21628420 TI - Telemedicine in the Solomon Islands: 2006 to 2009. AB - Telemedicine has been used in the Solomon Islands since 2000. We used quantitative and qualitative methods to examine telemedicine use in the Solomon Islands from January 2006 to June 2009. During the study period 66 telemedicine cases were submitted to the store and forward telemedicine system being used there. These included orthopaedic, oncology, cardiothoracic, infectious, congenital, gastroenterology and dermatology cases. Most cases (52%) were submitted by doctors at the National Referral Hospital (NRH) in Honiara. The majority of responses came from the NRH (27%). A final, firm recommendation regarding patient diagnosis and/or care was given for 46% of the cases. Interviews were conducted with 23 stakeholders in the Solomon Islands and in Australia to better understand the current and future use of telemedicine. The interviews identified the fragility of the Solomon Islands infrastructure, including the lack of training, as the largest barrier to the future use of telemedicine. The best use of telemedicine appears to be case sharing within the Solomon Islands, with connections to clinicians in other countries as a secondary benefit when particular expertise is required. PMID- 21628421 TI - Effectiveness and safety of a glucose data-filtering system with automatic response software to reduce the physician workload in managing type 2 diabetes. AB - We conducted a randomized controlled trial to investigate the efficacy and safety of a system for online glucose monitoring of type 2 diabetes patients. The software automatically filtered the self-monitored blood glucose data to reduce physicians' time. In the control group, the physicians had to contact the patients manually. We measured the time spent by physicians for online management in a 24-week study. Seventy-nine patients were recruited. The frequency of physicians' online monitoring of the patients was decreased by 55% in patients with HbA(1c) <= 6.5% and by 29% when HbA(1c) > 6.5% (P < 0.01). Physicians' log in time was reduced by 67% and 55% in patients with HbA(1c) <=6.5% and >6.5%, respectively (P <0.05). HbA(1c) levels were maintained at <6.5% during the study period in both groups. The study showed the efficacy and safety of the software for online communication in diabetes management. The results suggest that it could improve the cost-effectiveness of online communication systems and form the basis of future clinical applications. PMID- 21628422 TI - European perspective on the diagnosis and treatment of posthaemorrhagic ventricular dilatation. AB - BACKGROUND: Posthaemorrhagic ventricular dilatation (PHVD) is a serious complication of prematurity with subsequent disabilities. The diagnostic and therapeutic approaches to PHVD vary among neonatal centres. AIM: To gain more insight into the different diagnostic criteria and treatment policies on PHVD among neonatal intensive care units across Europe. Methods A PHVD questionnaire was designed and sent to neonatologists in 37 European centres. RESULTS: A response was obtained from 32/37 (86%) centres located in 17 European countries. An overall estimated incidence of 7% was reported for severe intraventricular haemorrhages (grades III or IV according to Papile) among premature neonates born below 30 weeks' gestation. Approximately half of these infants developed PHVD, of whom three-quarters required intervention. Ultrasound measurements of ventricular size were most commonly used to diagnose PHVD (94%). No consensus existed on which ventricular parameters needed to be enlarged and when to start treatment of PHVD. Early intervention (ie, initiated after the ventricular index (VI) exceeded the 97th percentile (p97) according to Levene) was provided in 8/32 centres (25%), whereas 23/32 centres (72%) first started therapy once the VI had crossed the p97+4 mm line and/or when neonates presented with a progressive increase in head circumference or with clinical symptoms of raised intracranial pressure. Wide variation was seen with respect to the applied therapy modalities for cerebrospinal fluid drainage. CONCLUSION: This survey shows that diagnostic and therapeutic approaches to neonates with PHVD vary considerably. Uniform diagnostic criteria would facilitate studies to assess optimal timing and mode of intervention. PMID- 21628423 TI - The non-receptor tyrosine kinase Lyn controls neutrophil adhesion by recruiting the CrkL-C3G complex and activating Rap1 at the leading edge. AB - Establishing new adhesions at the extended leading edges of motile cells is essential for stable polarity and persistent motility. Despite recent identification of signaling pathways that mediate polarity and chemotaxis in neutrophils, little is known about molecular mechanisms governing cell extracellular-matrix (ECM) adhesion in these highly polarized and rapidly migrating cells. Here, we describe a signaling pathway in neutrophils that is essential for localized integrin activation, leading edge attachment and persistent migration during chemotaxis. This pathway depends upon G(i)-protein mediated activation and leading edge recruitment of Lyn, a non-receptor tyrosine kinase belonging to the Src kinase family. We identified the small GTPase Rap1 as a major downstream effector of Lyn to regulate neutrophil adhesion during chemotaxis. Depletion of Lyn in neutrophil-like HL-60 cells prevented chemoattractant-induced Rap1 activation at the leading edge of the cell, whereas ectopic expression of Rap1 largely rescued the defects induced by Lyn depletion. Furthermore, Lyn controls spatial activation of Rap1 by recruiting the CrkL-C3G protein complex to the leading edge. Together, these results provide novel mechanistic insights into the poorly understood signaling network that controls leading edge adhesion during chemotaxis of neutrophils, and possibly other amoeboid cells. PMID- 21628424 TI - Increased ER-mitochondrial coupling promotes mitochondrial respiration and bioenergetics during early phases of ER stress. AB - Increasing evidence indicates that endoplasmic reticulum (ER) stress activates the adaptive unfolded protein response (UPR), but that beyond a certain degree of ER damage, this response triggers apoptotic pathways. The general mechanisms of the UPR and its apoptotic pathways are well characterized. However, the metabolic events that occur during the adaptive phase of ER stress, before the cell death response, remain unknown. Here, we show that, during the onset of ER stress, the reticular and mitochondrial networks are redistributed towards the perinuclear area and their points of connection are increased in a microtubule-dependent fashion. A localized increase in mitochondrial transmembrane potential is observed only in redistributed mitochondria, whereas mitochondria that remain in other subcellular zones display no significant changes. Spatial re-organization of these organelles correlates with an increase in ATP levels, oxygen consumption, reductive power and increased mitochondrial Ca2+ uptake. Accordingly, uncoupling of the organelles or blocking Ca2+ transfer impaired the metabolic response, rendering cells more vulnerable to ER stress. Overall, these data indicate that ER stress induces an early increase in mitochondrial metabolism that depends crucially upon organelle coupling and Ca2+ transfer, which, by enhancing cellular bioenergetics, establishes the metabolic basis for the adaptation to this response. PMID- 21628425 TI - Nuclear Chk1 prevents premature mitotic entry. AB - Chk1 inhibits the premature activation of the cyclin-B1-Cdk1. However, it remains controversial whether Chk1 inhibits Cdk1 in the centrosome or in the nucleus before the G2-M transition. In this study, we examined the specificity of the mouse monoclonal anti-Chk1 antibody DCS-310, with which the centrosome was stained. Conditional Chk1 knockout in mouse embryonic fibroblasts reduced nuclear but not centrosomal staining with DCS-310. In Chk1(+/myc) human colon adenocarcinoma (DLD-1) cells, Chk1 was detected in the nucleus but not in the centrosome using an anti-Myc antibody. Through the combination of protein array and RNAi technologies, we identified Ccdc-151 as a protein that crossreacted with DCS-310 on the centrosome. Mitotic entry was delayed by expression of the Chk1 mutant that localized in the nucleus, although forced immobilization of Chk1 to the centrosome had little impact on the timing of mitotic entry. These results suggest that nuclear but not centrosomal Chk1 contributes to correct timing of mitotic entry. PMID- 21628426 TI - Membrane organization and lipid rafts. AB - Cell membranes are composed of a lipid bilayer, containing proteins that span the bilayer and/or interact with the lipids on either side of the two leaflets. Although recent advances in lipid analytics show that membranes in eukaryotic cells contain hundreds of different lipid species, the function of this lipid diversity remains enigmatic. The basic structure of cell membranes is the lipid bilayer, composed of two apposing leaflets, forming a two-dimensional liquid with fascinating properties designed to perform the functions cells require. To coordinate these functions, the bilayer has evolved the propensity to segregate its constituents laterally. This capability is based on dynamic liquid-liquid immiscibility and underlies the raft concept of membrane subcompartmentalization. This principle combines the potential for sphingolipid-cholesterol self-assembly with protein specificity to focus and regulate membrane bioactivity. Here we will review the emerging principles of membrane architecture with special emphasis on lipid organization and domain formation. PMID- 21628427 TI - Quality control of mitochondrial proteostasis. AB - A decline in mitochondrial activity has been associated with aging and is a hallmark of many neurological diseases. Surveillance mechanisms acting at the molecular, organellar, and cellular level monitor mitochondrial integrity and ensure the maintenance of mitochondrial proteostasis. Here we will review the central role of mitochondrial chaperones and proteases, the cytosolic ubiquitin proteasome system, and the mitochondrial unfolded response in this interconnected quality control network, highlighting the dual function of some proteases in protein quality control within the organelle and for the regulation of mitochondrial fusion and mitophagy. PMID- 21628428 TI - Role of lipids in virus replication. AB - Viruses intricately interact with and modulate cellular membranes at several stages of their replication, but much less is known about the role of viral lipids compared to proteins and nucleic acids. All animal viruses have to cross membranes for cell entry and exit, which occurs by membrane fusion (in enveloped viruses), by transient local disruption of membrane integrity, or by cell lysis. Furthermore, many viruses interact with cellular membrane compartments during their replication and often induce cytoplasmic membrane structures, in which genome replication and assembly occurs. Recent studies revealed details of membrane interaction, membrane bending, fission, and fusion for a number of viruses and unraveled the lipid composition of raft-dependent and -independent viruses. Alterations of membrane lipid composition can block viral release and entry, and certain lipids act as fusion inhibitors, suggesting a potential as antiviral drugs. Here, we review viral interactions with cellular membranes important for virus entry, cytoplasmic genome replication, and virus egress. PMID- 21628429 TI - Protein kinase C: the "masters" of calcium and lipid. AB - The coordinated and physiological behavior of living cells in an organism critically depends on their ability to interact with surrounding cells and with the extracellular space. For this, cells have to interpret incoming stimuli, correctly process the signals, and produce meaningful responses. A major part of such signaling mechanisms is the translation of incoming stimuli into intracellularly understandable signals, usually represented by second messengers or second-messenger systems. Two key second messengers, namely the calcium ion and signaling lipids, albeit extremely different in nature, play an important and often synergistic role in such signaling cascades. In this report, we will shed some light on an entire family of protein kinases, the protein kinases C, that are perfectly designed to exactly decode these two second messengers in all of their properties and convey the signaling content to downstream processes within the cell. PMID- 21628431 TI - The influence of the local sequence environment on RNA loop structures. AB - RNA folding is assumed to be a hierarchical process. The secondary structure of an RNA molecule, signified by base-pairing and stacking interactions between the paired bases, is formed first. Subsequently, the RNA molecule adopts an energetically favorable three-dimensional conformation in the structural space determined mainly by the rotational degrees of freedom associated with the backbone of regions of unpaired nucleotides (loops). To what extent the backbone conformation of RNA loops also results from interactions within the local sequence context or rather follows global optimization constraints alone has not been addressed yet. Because the majority of base stacking interactions are exerted locally, a critical influence of local sequence on local structure appears plausible. Thus, local loop structure ought to be predictable, at least in part, from the local sequence context alone. To test this hypothesis, we used Random Forests on a nonredundant data set of unpaired nucleotides extracted from 97 X-ray structures from the Protein Data Bank (PDB) to predict discrete backbone angle conformations given by the discretized eta/theta-pseudo-torsional space. Predictions on balanced sets with four to six conformational classes using local sequence information yielded average accuracies of up to 55%, thus significantly better than expected by chance (17%-25%). Bases close to the central nucleotide appear to be most tightly linked to its conformation. Our results suggest that RNA loop structure does not only depend on long-range base-pairing interactions; instead, it appears that local sequence context exerts a significant influence on the formation of the local loop structure. PMID- 21628430 TI - Pseudouridine formation in archaeal RNAs: The case of Haloferax volcanii. AB - Pseudouridine (Psi), the isomer of uridine, is commonly found at various positions of noncoding RNAs of all organisms. Psi residues are formed by a number of single- or multisite specific Psi synthases, which generally act as stand alone proteins. In addition, in Eukarya and Archaea, specific ribonucleoprotein complexes, each containing a distinct box H/ACA guide RNA and four core proteins, can produce Psi at many sites of different cellular RNAs. Cbf5 is the core Psi synthase in these complexes. Using Haloferax volcanii as an archaeal model organism, we show that, contrary to eukaryotes, the Cbf5 homolog (HVO_2493) is not essential in this archaeon. The Cbf5-deleted strain of H. volcanii completely lacks Psi at positions 1940, 1942, 2605, and 2591 (Escherichia coli positions 1915, 1917, 2572, and 2586) of its 23S rRNA, and contains reduced steady-state levels of some box H/ACA RNAs. Archaeal Cbf5 is known to have tRNA Psi55 synthase activity in vitro but we could not confirm this activity in vivo in H. volcanii. Conversely, the Pus10 (previously PsuX) homolog (HVO_1979), which can produce tRNA Psi55, as well as Psi54 in vitro, is shown here to be essential in H. volcanii, whereas the corresponding tRNA Psi55 synthases, Pus4 and TruB, are not essential in yeast and E. coli, respectively. Finally, we demonstrate that HVO_1852, the TruA/Pus3 homolog, is responsible for the pseudouridylation of position 39 in H. volcanii tRNAs and that the corresponding gene is not essential. PMID- 21628432 TI - Zygotic amplification of secondary piRNAs during silkworm embryogenesis. AB - PIWI-interacting RNAs (piRNAs) are 23-30-nucleotide-long small RNAs that act as sequence-specific silencers of transposable elements in animal gonads. In flies, genetics and deep sequencing data have led to a hypothesis for piRNA biogenesis called the ping-pong cycle, where antisense primary piRNAs initiate an amplification loop to generate sense secondary piRNAs. However, to date, the process of the ping-pong cycle has never been monitored at work. Here, by large scale profiling of piRNAs from silkworm ovary and embryos of different developmental stages, we demonstrate that maternally inherited antisense-biased piRNAs trigger acute amplification of secondary sense piRNA production in zygotes, at a time coinciding with zygotic transcription of sense transposon mRNAs. These results provide on-site evidence for the ping-pong cycle. PMID- 21628433 TI - Fluorescent labeling of tRNA dihydrouridine residues: Mechanism and distribution. AB - Dihydrouridine (DHU) positions within tRNAs have long been used as sites to covalently attach fluorophores, by virtue of their unique chemical reactivity toward reduction by NaBH(4), their abundance within prokaryotic and eukaryotic tRNAs, and the biochemical functionality of the labeled tRNAs so produced. Interpretation of experiments employing labeled tRNAs can depend on knowing the distribution of dye among the DHU positions present in a labeled tRNA. Here we combine matrix-assisted laser desorption/ionization mass spectroscopy (MALDI-MS) analysis of oligonucleotide fragments and thin layer chromatography to resolve and quantify sites of DHU labeling by the fluorophores Cy3, Cy5, and proflavin in Escherichia coli tRNA(Phe) and E. coli tRNA(Arg). The MALDI-MS results led us to re-examine the precise chemistry of the reactions that result in fluorophore introduction into tRNA. We demonstrate that, in contrast to an earlier suggestion that has long been unchallenged in the literature, such introduction proceeds via a substitution reaction on tetrahydrouridine, the product of NaBH(4) reduction of DHU, resulting in formation of substituted tetrahydrocytidines within tRNA. PMID- 21628434 TI - Cervicogenic headache and onabotulinumtoxinA: where do we stand? PMID- 21628435 TI - Numbness matters: a clinical review of trigeminal neuropathy. AB - AIM: Trigeminal neuropathies are a group of clinical disorders that involve injury to primary first-order neurons within the trigeminal nerve. We review the spectrum of etiologies underlying both painful and non-painful trigeminal neuropathies, with attention to particularly dangerous processes that may elude the clinician in the absence of a meticulous evaluation. Complications and management issues specific to patients with trigeminal neuropathy are discussed. METHODS: Retrospective literature review. RESULTS: Facial or intraoral numbness, the hallmark of trigeminal neuropathy, may represent the earliest symptomology of malignancy or autoimmune connective tissue disease as sensory neurons are destroyed. Such numbness, especially if progressive, necessitates periodic evaluation and vigilance even years after presentation if no diagnosis can be made. CONCLUSIONS: In the routine evaluation of patients with facial pain, the clinician will inevitably be confronted with secondary pathology of the trigeminal nerves and nuclei. The appearance of numbness, even when pain continues to be the most pressing complaint, necessitates clinical assessment of the integrity of all aspects of the trigeminal pathways, which may also include neurophysiologic, radiographic, and laboratory evaluation. PMID- 21628436 TI - Development of visual contour integration in children with migraine without aura. AB - INTRODUCTION: As migraine attacks pose insult to cerebral circulation and ion homeostasis, migraine has the potential to interfere with the development of different brain structures, producing functional deficits. It is known that visual contour integration (CI) is a function with a protracted development. Therefore, we sought to establish whether migraine interferes with its development. METHODS: Forty-eight migraineurs (without aura) and 48 age- and sex matched controls participated in the study, divided into three cohorts by age. Stimuli were presented on cards with a contour consisting of Gabor patches embedded in random noise. Difficulty was varied by the manipulation of relative noise density. The task was to identify and show the contour. RESULTS: A significant difference was found between the performance of migraineurs and controls in the 10-14-year-old and 15-18-year-old cohorts (p < 0.05). Development between all three cohorts was significant in the control group (p < 0.017), while it was not significant in migraineurs between 6 and 14 years. Correlation between age and CI threshold was stronger in controls than in migraineurs. CONCLUSION: Children with paediatric migraine exhibited a less marked development in the Gabor patch-based CI task. PMID- 21628437 TI - Palinopsia in patients with migraine: a case-control study. AB - OBJECTIVES: This study was aimed at investigating the frequency of the visual phenomenon of palinopsia (visual perseveration) in patients with migraine. METHODS: We interviewed 63 patients with migraine with aura (MwA), 137 patients with migraine without aura (MwoA) and 226 sex-age-matched healthy control subjects using an ad hoc structured interview/questionnaire. The interview was divided into four classes of variables for statistical testing. RESULTS: Palinopsia occurred in 19/200 patients (9.5%); of them 10/63 had MwA and 9/137 MwoA (14.2% vs 6.6%, chi = 9.7, degrees of freedom = 1, p = 0.002). Patients with palinopsia had a significantly lower migraine attack frequency than those without this visual phenomenon (4.3 +/- 0.3 vs 14.4 +/- 0.2, z = 7.1, p < 0.0001). No healthy control subjects complained of palinopsia according to the structured interview/questionnaire. DISCUSSION: Palinopsia is probably under-diagnosed in patients with migraine. Further investigations are needed to assess whether migraineurs are particularly susceptible to the development of recurrent episodes of visual perseveration. PMID- 21628438 TI - A subjective cognitive impairment scale for migraine attacks. The MIG-SCOG: development and validation. AB - BACKGROUND: The burden of migraine is determined by impairment during attacks due to pain or non-painful symptoms such as cognitive symptoms. OBJECTIVE: Development of a questionnaire to measure self-reported subjective cognitive symptoms during migraine attacks. METHODS: Item generation was accomplished through structured patient interviews analysed by a panel of experts. A set of 43 candidate items was applied to consecutive migraine patients. Test construction with factor analysis retained nine items. Internal consistency was assessed with Cronbach's alpha and Spearman's rho, and convergent and construct validity by correlation to spontaneous cognitive complaints, and the 43-item and the Cognitive Failures Questionnaires. RESULTS: The nine-item Mig-SCog covers two domains, executive functions and language. Cronbach's alpha was 0.82. It correlates with spontaneous cognitive complaints (p < 0.001), the 43-item (rho = 0.69) and the Cognitive Failures Questionnaires (rho = 0.61). Test-retest reliability (Cohen's kappa) was 0.55. CONCLUSIONS: Mig-SCog is a valid, reliable, consistent working instrument of fast self-administration that quantifies subjective cognitive symptoms during migraine attacks. PMID- 21628439 TI - Sun-induced migraine attacks in an Arctic population. AB - AIMS: The main purpose of the study was to evaluate the impact of weather, and especially sun exposure, on migraine. METHODS: Data from a previous prospective 12-month diary study was compared with meteorological data. We retrospectively evaluated 1250 migraine attacks recorded by a group of 40 women with a mean age of 37.1 years who fulfilled the IHS criteria for migraine with and without aura. RESULTS: The patients reported more sun-induced migraine attacks on sunny days, but the total distribution of migraine attacks was constant throughout the year. Also, no seasonal variation of migraine, nor any relationships between weather parameters and onset of migraine attacks, were found. An analysis of a subgroup of patients with 'sun-induced' migraine showed a significant increase in frequency of migraine attacks in the summer compared to the winter (p = 0.04). CONCLUSION: This study confirms that sunlight might be a trigger for migraine, but a risk for increased impact of light on the total ailment of migraine headache should be searched for in a subgroup of sensitive migraineurs. PMID- 21628440 TI - Evaluation of olfaction in patients with migraine using an odour stick identification test. AB - AIMS: Peculiar characteristics of migraine headaches include the arousal of olfaction during the attacks and osmophobia. We performed an olfactory test to evaluate the association between olfaction and migraines. METHODS: We evaluated olfactory dysfunction in 80 migraine patients (31 experienced migraines with aura (MWA), 49 migraine without aura (MWOA)) and 30 healthy controls. Participants were assessed for concurrent osmophobia. Olfaction was evaluated using an odour stick identification test (OSIT), in which participants were asked to identify various odours during a migraine-free period. The degree of offensiveness of each odour was also evaluated. RESULTS: Sixty-three percent of migraine patients were found to have concurrent osmophobia (MWA 71%; MWOA 57%). The percentages of migraine patients and controls who correctly identified test odours were 91% (92%, MWA; 89%, MWOA) and 92%, respectively. Perfume, rose and Japanese cypress odours were more offensive to migraine patients than to controls. All test odours were found to be more offensive to MWA than to MWOA patients. CONCLUSIONS: The OSIT showed certain odours to be highly offensive to migraine patients even when they were not experiencing migraine headaches. More attention should be paid to odours that are perceived to be offensive by migraine patients, particularly those with MWA. PMID- 21628441 TI - Morning headache in habitual snorers. PMID- 21628442 TI - Saccade behaviour in migraine patients. AB - BACKGROUND: Voxel-based morphometry studies in migraine patients showed significant grey matter volume reduction in regions involved in the control of saccadic eye movements. We hypothesized that these changes would be reflected in dysfunctional saccadic behaviour. METHODS: Saccades were recorded by infrared oculography using three different paradigms (pro-saccade with gap, pro-saccade overlap and anti-saccade with gap). We compared the results for migraine patients (n = 80) with those for controls (n = 87). RESULTS: No significant differences were found between migraine patients with (n = 46) and without (n = 34) aura. Migraine patients showed a saccadic behaviour that differed from controls in three respects. In migraine patients, the latencies in the pro-saccade with gap paradigm were borderline significantly longer. Moreover, in both the pro-saccade with gap and the pro-saccade overlap paradigm we observed a larger intra individual variation of the latency in migraine patients. However, the biggest difference was that the patients who received migraine prophylactic therapy made significantly more anti-saccade errors in the anti-saccade with gap paradigm, suggesting that inhibitory saccade control is impaired in migraine patients depending on the severity of the migraine. CONCLUSION: We suggest a deficient inhibitory control, reflecting an executive dysfunction in the dorsolateral prefrontal cortex or a dysfunction in the cingulate cortex, is present in migraine patients. PMID- 21628443 TI - Deep brain stimulation of the posterior hypothalamic area in intractable short lasting unilateral neuralgiform headache with conjunctival injection and tearing (SUNCT). AB - BACKGROUND: SUNCT (short-lasting unilateral neuralgiform headache with conjunctival injection and tearing) is a rare syndrome characterized by the sudden onset of excruciating unilateral periorbital pain that is accompanied by conjunctival injection and lacrimation or further autonomic signs. Similar to patients with chronic cluster headache, Leone and Lyons showed a beneficial effect of deep brain stimulation of the posterior hypothalamic region in two patients with a chronic SUNCT. CASE: Here, we present the case of a man with a chronic SUNCT responding to deep brain stimulation of the posterior hypothalamic area. CONCLUSION: This case supports the idea of a central origin of SUNCT and shows that deep brain stimulation of the hypothalamic region can be effective in the treatment of the chronic form of this rare disorder. PMID- 21628444 TI - Epidemiology of medication overuse headache in the general Swedish population. AB - AIMS: The aim was to estimate the prevalence of medication overuse headache (MOH) in Sweden and to analyze the occurrence of this disorder in different population groups. METHODS: A total of 44,300 randomly selected individuals (55% women), aged 15 years and above, were interviewed in a national telephone survey, using a standardized questionnaire including the International Headache Society criteria for MOH and questions about sociodemographic factors, headache history and medication use. RESULTS: In Sweden, 3.2% (95% confidence interval (CI) 3.1-3.4), n = 1428) suffer from chronic daily headache (CDH) and out of those, 56% (n = 799) have MOH. The prevalence of MOH is 1.8% (95% CI 1.7-1.9). The mean age of onset was higher among men than women as well as among those with tension-type headache as primary headache compared to those who originally had migraine. A multivariate analysis showed that socioeconomic factors such as having a low level of education and/or a low household income were associated with MOH. CONCLUSIONS: This is the first Swedish population-based study of MOH and we conclude that MOH is a significant public health problem in Sweden, as it is in other parts of the world. PMID- 21628445 TI - Tumor exploits alternative strategies to achieve vascularization. AB - Neoangiogenesis is crucial for solid tumor growth and invasion, as the vasculature provides metabolic support and access to the circulation. Current antiangiogenic therapies have been designed on the assumption that endothelial cells forming the tumor vasculature exhibit genetic stability. Recent studies demonstrate that this is not the case. Tumor endothelial cells possess a distinct phenotype, differing from normal endothelial cells at both molecular and functional levels. This challenges the concept that tumor angiogenesis exclusively depends on normal endothelial cell recruitment from the surrounding vascular network. Indeed, recent data suggest alternative strategies for tumor vascularization. It has been reported that tumor vessels may derive from an intratumor embryonic-like vasculogenesis. This condition might be due to differentiation of normal stem and progenitor cells of hematopoietic origin or resident in tissues. Cancer stem cells may also participate in tumor vasculogenesis by virtue of their stem and progenitor cell properties. Finally, normal endothelial cells might be reprogrammed to a proangiogenic or dedifferentiated phenotype by genetic information transmitted from the tumor trough apoptotic bodies, or following mRNA and microRNA transfer by exosomes and microvesicles. In this review, we discuss the different aspects of intratumor angiogenesis and vasculogenesis, the known mechanisms involved, and the possible implications for the response to antiangiogenic therapy. PMID- 21628446 TI - Angioprotectin: an angiotensin II-like peptide causing vasodilatory effects. AB - The family of angiotensin peptides has been steadily growing in recent years. Most are fragments of angiotensin II (Ang II) with different affinities to the known angiotensin receptors. Here, we describe a novel endogenous Ang II-like octapeptide in plasma from healthy humans and patients with end-stage renal failure, which acts as a stronger agonist at Mas receptors than Ang 1-7. Chromatographic purification and structural analysis by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI-TOF/TOF) revealed an Ang II-like octapeptide, angioprotectin, with the sequence Pro-Glu-Val-Tyr-Ile His-Pro-Phe, which differs from Ang II in Pro1 and Glu2 instead of Asp1 and Arg2. Pro-Glu-Val-Tyr-Ile-His-Pro-Phe in angioprotectin is most likely generated enzymatically from Ang II. Angioprotectin antagonized the contractile actions of Ang II on rat aortic rings. The physiological antagonism of vasoconstrictor actions of Ang II by angioprotectin is mediated by the Mas receptor. Angioprotectin has a stronger affinity to the Mas receptor than Ang-1-7. Plasma concentrations were ~15% of plasma Ang II concentrations in healthy volunteers and up to 50% in patients with renal failure. A commercially available Ang II antibody did not discriminate between angioprotectin and Ang II; thus, angioprotectin can contribute to Ang II concentrations measured by antibody-based assays. This novel peptide is likely to be a relevant component of the human renin-angiotensin-system. PMID- 21628447 TI - Lipoxin A4 and benzo-lipoxin A4 attenuate experimental renal fibrosis. AB - Unresolved inflammation underlies the development of fibrosis and organ failure. Here, we investigate the potential of the proresolving eicosanoid lipoxinA4 (LXA4) and its synthetic analog benzo-LXA4 to prophylactically modulate fibrotic and inflammatory responses in a model of early renal fibrosis, unilateral ureteric obstruction (UUO). Male Wistar rats (Animalia, Chordata, Rattus norvegicus) were injected intravenously with vehicle (0.1% ethanol), LXA4 (45 MUg/250-g rat), or benzo-LXA4 (15 MUg/250-g rat) 15 min prior to surgery and sacrificed 3 d postligation. Renal gene and protein expression, collagen deposition, macrophage infiltration, and apoptosis were analyzed using manipulated kidneys from sham operations as control. Lipoxins (LXs) attenuated collagen deposition and renal apoptosis (P<0.05) and shifted the inflammatory milieu toward resolution, inhibiting TNF-alpha and IFN-gamma expression, while stimulating proresolving IL-10. LXs attenuated UUO-induced activation of MAP kinases, Akt, and Smads (P<0.05) in injured kidneys. We explored whether the underlying mechanism reflected LX-induced modulation of fibroblast activation. Using cultured rat renal NRK-49F fibroblasts, we report that LXA4 (1 nM) inhibits TGF-beta1 (10 ng/ml)-induced activation of Smad2 and MAP-kinases (P<0.05), and furthermore, LXA4 reduced TGF-beta1-stimulated PAI-1 luciferase activation (P<0.05) relative to vehicle-stimulated cells. We propose that LXs may represent a potentially useful and novel therapeutic strategy for consideration in the context of renal fibrosis. PMID- 21628448 TI - Defective adenosine-stimulated cAMP production in cystic fibrosis airway epithelia: a novel role for CFTR in cell signaling. AB - Adenosine (ADO) is an extracellular signaling molecule that is an important regulator of innate lung defense. On binding ADO, the A2B receptor (A2BR) stimulates cAMP production to activate the CFTR Cl(-) channel, increase ciliary beating, and initiate cytokine secretion. We tested the hypothesis that CFTR served as a positive regulator of the A2BRs. We found that A2BR and CFTR coimmunoprecipitated. They also underwent ADO-dependent Forster resonance energy transfer (FRET), which increased from 5% in the absence of agonist to 18% with 100 MUM ADO (EC50 1.7 MUM), suggesting that they dynamically associate in the plasma membrane. In contrast, despite colocalization, no FRET was observed between CFTR and GAP43. The interaction between A2BR and CFTR had some specificity: A2BR-stimulated but not forskolin-stimulated cAMP production was ~50% greater in the presence of CFTR, due to a CFTR-dependent increase in plasma membrane A2BR levels. These CFTR-dependent increases in A2BR levels and cAMP production resulted in significantly enhanced ciliary beating and increased cytokine secretion in normal compared to cystic fibrosis airway epithelia. Thus, we hypothesize that CFTR regulates A2BR levels in the plasma membrane to modulate cell signaling and to enhance selective components of the innate lung defense system. PMID- 21628449 TI - Cell type-specific DNA methylation at intragenic CpG islands in the immune system. AB - Human and mouse genomes contain a similar number of CpG islands (CGIs), which are discrete CpG-rich DNA sequences associated with transcription start sites. In both species, ~50% of all CGIs are remote from annotated promoters but, nevertheless, often have promoter-like features. To determine the role of CGI methylation in cell differentiation, we analyzed DNA methylation at a comprehensive CGI set in cells of the mouse hematopoietic lineage. Using a method that potentially detects ~33% of genomic CpGs in the methylated state, we found that large differences in gene expression were accompanied by surprisingly few DNA methylation changes. There were, however, many DNA methylation differences between hematopoietic cells and a distantly related tissue, brain. Altered DNA methylation in the immune system occurred predominantly at CGIs within gene bodies, which have the properties of cell type-restricted promoters, but infrequently at annotated gene promoters or CGI flanking sequences (CGI "shores"). Unexpectedly, elevated intragenic CGI methylation correlated with silencing of the associated gene. Differentially methylated intragenic CGIs tended to lack H3K4me3 and associate with a transcriptionally repressive environment regardless of methylation state. Our results indicate that DNA methylation changes play a relatively minor role in the late stages of differentiation and suggest that intragenic CGIs represent regulatory sites of differential gene expression during the early stages of lineage specification. PMID- 21628450 TI - Genome-wide identification of conserved regulatory function in diverged sequences. AB - Plasticity of gene regulatory encryption can permit DNA sequence divergence without loss of function. Functional information is preserved through conservation of the composition of transcription factor binding sites (TFBS) in a regulatory element. We have developed a method that can accurately identify pairs of functional noncoding orthologs at evolutionarily diverged loci by searching for conserved TFBS arrangements. With an estimated 5% false-positive rate (FPR) in approximately 3000 human and zebrafish syntenic loci, we detected approximately 300 pairs of diverged elements that are likely to share common ancestry and have similar regulatory activity. By analyzing a pool of experimentally validated human enhancers, we demonstrated that 7/8 (88%) of their predicted functional orthologs retained in vivo regulatory control. Moreover, in 5/7 (71%) of assayed enhancer pairs, we observed concordant expression patterns. We argue that TFBS composition is often necessary to retain and sufficient to predict regulatory function in the absence of overt sequence conservation, revealing an entire class of functionally conserved, evolutionarily diverged regulatory elements that we term "covert." PMID- 21628451 TI - A reduced representation approach to population genetic analyses and applications to human evolution. AB - Second-generation sequencing technologies allow surveys of sequence variation on an unprecedented scale. However, despite the rapid decrease in sequencing costs, collecting whole-genome sequence data on a population scale is still prohibitive for many laboratories. We have implemented an inexpensive, reduced representation protocol for preparing resequencing targets, and we have developed the analytical tools necessary for making population genetic inferences. This approach can be applied to any species for which a draft or complete reference genome sequence is available. The new tools we have developed include methods for aligning reads, calling genotypes, and incorporating sample-specific sequencing error rates in the estimate of evolutionary parameters. When applied to 19 individuals from a total of 18 human populations, our approach allowed sampling regions that are largely overlapping across individuals and that are representative of the entire genome. The resequencing data were used to test the serial founder model of human dispersal and to estimate the time of the Out of Africa migration. Our results also represent the first attempt to provide a time frame for the colonization of Australia based on large-scale resequencing data. PMID- 21628452 TI - Pervasive haplotypic variation in the spliceo-transcriptome of the human major histocompatibility complex. AB - The human major histocompatibility complex (MHC) on chromosome 6p21 is a paradigm for genomics, showing remarkable polymorphism and striking association with immune and non-immune diseases. The complex genomic landscape of the MHC, notably strong linkage disequilibrium, has made resolving causal variants very challenging. A promising approach is to investigate gene expression levels considered as tractable intermediate phenotypes in mapping complex diseases. However, how transcription varies across the MHC, notably relative to specific haplotypes, remains unknown. Here, using an original hybrid tiling and splice junction microarray that includes alternate allele probes, we draw the first high resolution strand-specific transcription map for three common MHC haplotypes (HLA A1-B8-Cw7-DR3, HLA-A3-B7-Cw7-DR15, and HLA-A26-B18-Cw5-DR3-DQ2) strongly associated with autoimmune diseases including type 1 diabetes, systemic lupus erythematosus, and multiple sclerosis. We find that haplotype-specific differences in gene expression are common across the MHC, affecting 96 genes (46.4%), most significantly the zing finger protein gene ZFP57. Differentially expressed probes are correlated with polymorphisms between haplotypes, consistent with cis effects that we directly demonstrate for ZFP57 in a cohort of healthy volunteers (P = 1.2 * 10(-14)). We establish that alternative splicing is significantly more frequent in the MHC than genome-wide (72.5% vs. 62.1% of genes, P <= 1 * 10(-4)) and shows marked haplotypic differences. We also unmask novel and abundant intergenic transcription involving 31% of transcribed blocks identified. Our study reveals that the renowned MHC polymorphism also manifests as transcript diversity, and our novel haplotype-based approach marks a new step toward identification of regulatory variants involved in the control of MHC associated phenotypes and diseases. PMID- 21628453 TI - Normal fur development and sebum production depends on fatty acid 2-hydroxylase expression in sebaceous glands. AB - 2-Hydroxylated fatty acid (HFA)-containing sphingolipids are abundant in mammalian skin and are believed to play a role in the formation of the epidermal barrier. Fatty acid 2-hydroxylase (FA2H), required for the synthesis of 2 hydroxylated sphingolipids in various organs, is highly expressed in skin, and previous in vitro studies demonstrated its role in the synthesis of HFA sphingolipids in human keratinocytes. Unexpectedly, however, mice deficient in FA2H did not show significant changes in their epidermal HFA sphingolipids. Expression of FA2H in murine skin was restricted to the sebaceous glands, where it was required for synthesis of 2-hydroxylated glucosylceramide and a fraction of type II wax diesters. Absence of FA2H resulted in hyperproliferation of sebocytes and enlarged sebaceous glands during hair follicle morphogenesis and anagen (active growth phase) in adult mice. This was accompanied by a significant up-regulation of the epidermal growth factor receptor ligand epigen in sebocytes. Loss of FA2H significantly altered the composition and physicochemical properties of sebum, which often blocked the hair canal, apparently causing a delay in the hair fiber exit. Furthermore, mice lacking FA2H displayed a cycling alopecia with hair loss in telogen. These results underline the importance of the sebaceous glands and suggest a role of specific sebaceous gland or sebum lipids, synthesized by FA2H, in the hair follicle homeostasis. PMID- 21628454 TI - Disruption of a nuclear NFATc2 protein stabilization loop confers breast and pancreatic cancer growth suppression by zoledronic acid. AB - The aminobisphosphonate zoledronic acid has elicited significant attention due to its remarkable anti-tumoral activity, although its detailed mechanism of action remains unclear. Here, we demonstrate the existence of a nuclear GSK-3beta-NFATc2 stabilization pathway that promotes breast and pancreatic cancer growth in vitro and in vivo and serves as a bona fide target of zoledronic acid. Specifically, the serine/threonine kinase GSK-3beta stabilizes nuclear NFATc2 through phosphorylation of the serine-rich SP2 domain, thus protecting the transcription factor from E3-ubiquitin ligase HDM2-mediated proteolysis. Zoledronic acid disrupts this NFATc2 stabilization pathway through two mechanisms, namely GSK 3beta inhibition and induction of HDM2 activity. Upon nuclear accumulation, HDM2 targets unphosphorylated NFATc2 for ubiquitination at acceptor lysine residues Lys-684/Lys-897 and hence labels the factor for subsequent proteasomal degradation. Conversely, mutagenesis-induced constitutive serine phosphorylation (Ser-215, Ser-219, and Ser-223) of the SP2 domain prevents NFATc2 from HDM2 mediated ubiquitination and degradation and consequently rescues cancer cells from growth suppression by zoledronic acid. In conclusion, this study demonstrates a critical role of the GSK-3beta-HDM2 signaling loop in the regulation of NFATc2 protein stability and growth promotion and suggests that double targeting of this pathway is responsible, at least to a significant part, for the potent and reliable anti-tumoral effects of zoledronic acid. PMID- 21628455 TI - Increasing hydrophobicity or disulfide bridging at the factor VIII A1 and C2 domain interface enhances procofactor stability. AB - Factor VIII (FVIII) consists of a heavy (A1A2B domains) and light chain (A3C1C2 domains), whereas the contiguous A1A2 domains are separate subunits in the cofactor, FVIIIa. FVIII x-ray structures show close contacts between A1 and C2 domains. To explore the role of this region in FVIII(a) stability, we generated a variant containing a disulfide bond between A1 and C2 domains by mutating Arg-121 and Leu-2302 to Cys (R121C/L2302C) and a second variant with a bulkier hydrophobic group (A108I) to better occupy a cavity between A1 and C2 domains. Disulfide bonding in the R121C/L2302C variant was >90% efficient as judged by Western blots. Binding affinity between the A108I A1 and A3C1C2 subunits was increased ~3.7-fold in the variant as compared with WT as judged by changes in fluorescence of acrylodan-labeled A1 subunits. FVIII thermal and chemical stability were monitored following rates of loss of FVIII activity at 57 degrees C or in guanidinium by factor Xa generation assays. The rate of decay of FVIIIa activity was monitored at 23 degrees C following activation by thrombin. Both R121C/L2302C and A108I variants showed up to ~4-fold increases in thermal stability but minimal improvements in chemical stability. The purified A1 subunit of A108I reconstituted with the A3C1C2 subunit showed an ~4.6-fold increase in thermal stability, whereas reconstitution of the variant A1 with a truncated A3C1 subunit showed similar stability values as compared with WT A1. Together, these results suggest that altering contacts at this A1-C2 junction by covalent modification or increasing hydrophobicity increases inter-chain affinity and functionally enhances FVIII stability. PMID- 21628456 TI - The sterol-sensing domain (SSD) directly mediates signal-regulated endoplasmic reticulum-associated degradation (ERAD) of 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase isozyme Hmg2. AB - The sterol-sensing domain (SSD) is a conserved motif in membrane proteins responsible for sterol regulation. Mammalian proteins SREBP cleavage-activating protein (SCAP) and HMG-CoA reductase (HMGR) both possess SSDs required for feedback regulation of sterol-related genes and sterol synthetic rate. Although these two SSD proteins clearly sense sterols, the range of signals detected by this eukaryotic motif is not clear. The yeast HMG-CoA reductase isozyme Hmg2, like its mammalian counterpart, undergoes endoplasmic reticulum (ER)-associated degradation that is subject to feedback control by the sterol pathway. The primary degradation signal for yeast Hmg2 degradation is the 20-carbon isoprene geranylgeranyl pyrophosphate, rather than a sterol. Nevertheless, the Hmg2 protein possesses an SSD, leading us to test its role in feedback control of Hmg2 stability. We mutated highly conserved SSD residues of Hmg2 and evaluated regulated degradation. Our results indicated that the SSD was required for sterol pathway signals to stimulate Hmg2 ER-associated degradation and was employed for detection of both geranylgeranyl pyrophosphate and a secondary oxysterol signal. Our data further indicate that the SSD allows a signal-dependent structural change in Hmg2 that promotes entry into the ER degradation pathway. Thus, the eukaryotic SSD is capable of significant plasticity in signal recognition or response. We propose that the harnessing of cellular quality control pathways to bring about feedback regulation of normal proteins is a unifying theme for the action of all SSDs. PMID- 21628457 TI - Multiple BH3 mimetics antagonize antiapoptotic MCL1 protein by inducing the endoplasmic reticulum stress response and up-regulating BH3-only protein NOXA. AB - BH3 mimetics are small molecules designed or discovered to mimic the binding of BH3-only proteins to the hydrophobic groove of antiapoptotic BCL2 proteins. The selectivity of these molecules for BCL2, BCL-X(L), or MCL1 has been established in vitro; whether they inhibit these proteins in cells has not been rigorously investigated. In this study, we used a panel of leukemia cell lines to assess the ability of seven putative BH3 mimetics to inhibit antiapoptotic proteins in a cell-based system. We show that ABT-737 is the only BH3 mimetic that inhibits BCL2 as assessed by displacement of BAD and BIM from BCL2. The other six BH3 mimetics activate the endoplasmic reticulum stress response inducing ATF4, ATF3, and NOXA, which can then bind to and inhibit MCL1. In most cancer cells, inhibition of one antiapoptotic protein does not acutely induce apoptosis. However, by combining two BH3 mimetics, one that inhibits BCL2 and one that induces NOXA, apoptosis is induced within 6 h in a BAX/BAK-dependent manner. Because MCL1 is a major mechanism of resistance to ABT-737, these results suggest a novel strategy to overcome this resistance. Our findings highlight a novel signaling pathway through which many BH3 mimetics inhibit MCL1 and suggest the potential use of these agents as adjuvants in combination with various chemotherapy strategies. PMID- 21628458 TI - Tumor necrosis factor alpha-mediated induction of interleukin 17C in human keratinocytes is controlled by nuclear factor kappaB. AB - IL-17C is a member of the IL-17 family of cytokines. The expression of IL-17C has been demonstrated to be strongly induced by TNFalpha in human keratinocytes, and recently the level of IL-17C was found to be increased in the inflammatory skin disease psoriasis. However, little is known about the molecular mechanisms involved in the regulation of IL-17C. Here, we show that pretreatment of cultured human keratinocytes with the inhibitor of kappaB kinase 2 inhibitor, SC-514, resulted in a significant reduction in both IL-17C mRNA and protein expression, indicating the significance of this pathway in the regulation of IL-17C. NF kappaB binding sites were identified upstream from the IL-17C gene, and by electrophoretic mobility shift assay NF-kappaB was shown to bind to all three identified binding sites. Moreover, NF-kappaB binding to these sites was inducible by TNFalpha. Supershift analysis revealed binding of the NF-kappaB subunits p65 and p50 to all three NF-kappaB binding sites. To determine the contribution of NF-kappaB in IL-17C expression, we conducted luciferase gene reporter experiments and demonstrated that a 3204-bp promoter fragment of IL-17C containing three putative NF-kappaB binding sites was strongly activated by TNFalpha. Interestingly, mutations of the three NF-kappaB binding sites revealed that one specific NF-kappaB binding site was crucial for the TNFalpha-mediated IL 17C induction because mutation of this specific site completely abolished TNFalpha-induced IL-17C promoter activation. We conclude that the activation of NF-kappaB (p65/p50) is crucial for the TNFalpha-induced stimulation of IL-17C expression in human keratinocytes. PMID- 21628459 TI - Prenylated c17orf37 induces filopodia formation to promote cell migration and metastasis. AB - Post-translational modification by covalent attachment of isoprenoid lipids (prenylation) regulates the functions and biological activities of several proteins implicated in the oncogenic transformation and metastatic progression of cancer. The largest group of prenylated proteins contains a CAAX motif at the C terminal that serves as a substrate for a series of post-translational modifications that convert these otherwise hydrophilic proteins to lipidated proteins, thus facilitating membrane association. C17orf37 (chromosome 17 open reading frame 37), also known as C35/Rdx12/MGC14832, located in the 17q12 amplicon, is overexpressed in human cancer, and its expression correlates with the migratory and invasive phenotype of cancer cells. Here we show that C17orf37 contains a functional CAAX motif and is post-translationally modified by protein geranylgeranyltransferase-I (GGTase-I). Geranylgeranylation of C17orf37 at the CAAX motif facilitates association of the protein to the inner leaflet of plasma membrane, enhances migratory phenotype of cells by inducing increased filopodia formation, and potentiates directional migration. A prenylation-deficient mutant of C17orf37 is functionally inactive and fails to trigger dissemination of tail vein-injected cells in a mouse model of metastasis. These findings demonstrate that prenylation is required for the function of the C17orf37 protein in cancer cells and imply that the post-translational modification may functionally regulate metastatic progression of disease. PMID- 21628460 TI - TRIM32 protein sensitizes cells to tumor necrosis factor (TNFalpha)-induced apoptosis via its RING domain-dependent E3 ligase activity against X-linked inhibitor of apoptosis (XIAP). AB - TRIM32, which belongs to the tripartite motif (TRIM) protein family, has the RING finger, B-box, and coiled-coil domain structures common to this protein family, along with an additional NHL domain at the C terminus. TRIM32 reportedly functions as an E3 ligase for actin, a protein inhibitor of activated STAT y (PIASy), dysbindin, and c-Myc, and it has been associated with diseases such as muscular dystrophy and epithelial carcinogenesis. Here, we identify a new substrate of TRIM32 and propose a mechanism through which TRIM32 might regulate apoptosis. Our overexpression and knockdown experiments demonstrate that TRIM32 sensitizes cells to TNFalpha-induced apoptosis. The RING domain is necessary for this pro-apoptotic function of TRM32 as well as being responsible for its E3 ligase activity. TRIM32 colocalizes and directly interacts with X-linked inhibitor of apoptosis (XIAP), a well known cancer therapeutic target, through its coiled-coil and NHL domains. TRIM32 overexpression enhances XIAP ubiquitination and subsequent proteasome-mediated degradation, whereas TRIM32 knockdown has the opposite effect, indicating that XIAP is a substrate of TRIM32. In vitro reconstitution assay reveals that XIAP is directly ubiquitinated by TRIM32. Our novel results collectively suggest that TRIM32 sensitizes TNFalpha induced apoptosis by antagonizing XIAP, an anti-apoptotic downstream effector of TNFalpha signaling. This function may be associated with TRIM32-mediated tumor suppressive mechanism. PMID- 21628461 TI - C termini of proteasomal ATPases play nonequivalent roles in cellular assembly of mammalian 26 S proteasome. AB - The 26 S proteasome comprises two multisubunit subcomplexes as follows: 20 S proteasome and PA700/19 S regulatory particle. The cellular mechanisms by which these subcomplexes assemble into 26 S proteasome and the molecular determinants that govern the assembly process are poorly defined. Here, we demonstrate the nonequivalent roles of the C termini of six AAA subunits (Rpt1-Rpt6) of PA700 in 26 S proteasome assembly in mammalian cells. The C-terminal HbYX motif (where Hb is a hydrophobic residue, Y is tyrosine, and X is any amino acid) of each of two subunits, Rpt3 and Rpt5, but not that of a third subunit Rpt2, was essential for assembly of 26 S proteasome. The C termini of none of the three non-HbYX motif Rpt subunits were essential for cellular 26 S proteasome assembly, although deletion of the last three residues of Rpt6 destabilized the 20 S-PA700 interaction. Rpt subunits defective for assembly into 26 S proteasome due to C terminal truncations were incorporated into intact PA700. Moreover, intact PA700 accumulated as an isolated subcomplex when cellular 20 S proteasome content was reduced by RNAi. These results indicate that 20 S proteasome is not an obligatory template for assembly of PA700. Collectively, these results identify specific structural elements of two Rpt subunits required for 26 S proteasome assembly, demonstrate that PA700 can be assembled independently of the 20 S proteasome, and suggest that intact PA700 is a direct intermediate in the cellular pathway of 26 S proteasome assembly. PMID- 21628462 TI - Kallikrein-related peptidase 12 hydrolyzes matricellular proteins of the CCN family and modifies interactions of CCN1 and CCN5 with growth factors. AB - Kallikrein-related peptidases (KLKs) are an emerging group of secreted serine proteases involved in several physiological and pathological processes. We used a degradomic approach to identify potential substrates of KLK12. MDA-MB-231 cells were treated either with KLK12 or vehicle control, and the proteome of the overlying medium was analyzed by mass spectrometry. CCN1 (cyr61, ctgf, nov) was among the proteins released by the KLK12-treated cells, suggesting that KLK12 might be responsible for the shedding of this protein from the cell surface. Fragmentation of CCN1 by KLK12 was further confirmed in vitro, and the main cleavage site was localized in the hinge region between the first and second half of the recombinant protein. KLK12 can target all six members of the CCN family at different proteolytic sites. Limited proteolysis of CCNs (cyr61, ctgf, nov) was also observed in the presence of other members of the KLK family, such as KLK1, KLK5, and KLK14, whereas KLK6, KLK11, and KLK13 were unable to fragment CCNs. Because KLK12 seems to have a role in angiogenesis, we investigated the relations between KLK12, CCNs, and several factors known to be involved in angiogenesis. Solid phase binding assays showed that fragmentation of CCN1 or CCN5 by KLK12 prevents VEGF(165) binding, whereas it also triggers the release of intact VEGF and BMP2 from the CCN complexes. The KLK12-mediated release of TGF-beta1 and FGF 2, either as intact or truncated forms, was found to be concentration-dependent. These findings suggest that KLK12 may indirectly regulate the bioavailability and activity of several growth factors through processing of their CCN binding partners. PMID- 21628463 TI - The rate of interleukin-1beta secretion in different myeloid cells varies with the extent of redox response to Toll-like receptor triggering. AB - Human myeloid cells activate the NLRP3 inflammasome and secrete interleukin (IL) 1beta in response to various Toll-like receptor (TLR) ligands, but the rate of secretion is much higher in primary human monocytes than in cultured macrophages or THP-1 cells. The different myeloid cells also display different redox status under resting conditions and redox response to TLR activation. Resting monocytes display a balanced redox state, with low production of reactive oxygen species (ROS) and antioxidants. TLR engagement induces an effective redox response with increased ROS generation followed by a sustained antioxidant response, parallelled by efficient IL-1beta secretion. Drugs blocking ROS production or the antioxidant response prevent the secretion of mature IL-1beta but not the biosynthesis of pro-IL-1beta, indicating that redox remodeling is responsible for IL-1beta processing and release. Unlike monocytes, THP-1 cells and cultured macrophages have up-regulated antioxidant systems that buffer the oxidative hit provided by TLR triggering and suppress the consequent redox response. This aborted redox remodeling is paralleled by low efficiency IL-1beta processing and secretion. High doses (5 mM) of H(2)O(2) overcome the high antioxidant capacity of THP-1 cells, restore an efficient redox response, and increase the rate of IL 1beta secretion. Together these data indicate that a tightly controlled redox homeostasis in resting cells is a prerequisite for a robust redox response to TLR ligands, in turn necessary for the efficient inflammasome activation. Inflammasome activation by bacterial DNA is not modulated by redox responses, suggesting that redox-dependent regulation of IL-1beta secretion is restricted to some inflammasomes including NLRP3 but excluding AIM-2. PMID- 21628464 TI - Optogenetic control of motor coordination by Gi/o protein-coupled vertebrate rhodopsin in cerebellar Purkinje cells. AB - G protein-coupled receptors are involved in the modulation of complex neuronal networks in the brain. To investigate the impact of a cell-specific G(i/o) protein-mediated signaling pathway on brain function, we created a new optogenetic mouse model in which the G(i/o) protein-coupled receptor vertebrate rhodopsin can be cell-specifically expressed with the aid of Cre recombinase. Here we use this mouse model to study the functional impact of G(i/o) modulation in cerebellar Purkinje cells (PCs). We show that in vivo light activation of vertebrate rhodopsin specifically expressed in PCs reduces simple spike firing that is comparable with the reduction in firing observed for the activation of cerebellar G(i/o)-coupled GABA(B) receptors. Notably, the light exposure of the cerebellar vermis in freely moving mice changes the motor behavior. Thus, our studies directly demonstrate that spike modulation via G(i/o)-mediated signaling in cerebellar PCs affects motor coordination and show a new promising approach for studying the physiological function of G protein-coupled receptor-mediated signaling in a cell type-specific manner. PMID- 21628465 TI - Toll-like receptor-4 (TLR4) down-regulates microRNA-107, increasing macrophage adhesion via cyclin-dependent kinase 6. AB - Toll-like receptors (TLRs) modulate the expression of multiple microRNAs (miRNAs). Here, we report the down-regulation of miR-107 by TLR4 in multiple cell types. The miR-107 sequence occurs in an intron within the sequence encoding the gene for pantothenate kinase 1alpha (PanK1alpha), which is regulated by the transcription factor peroxisome proliferator-activating receptor alpha (PPAR alpha). PanK1alpha is also decreased in response to lipopolysaccharide (LPS). The effect on both miR-107 and PanK1alpha is consistent with a decrease in PPAR-alpha expression. We have found that the putative miR-107 target cyclin-dependent kinase 6 (CDK6) expression is increased by TLR4 as a result of the decrease in miR-107. This effect is required for increased adhesion of macrophages in response to LPS, and CDK6-deficient mice are resistant to the lethal effect of LPS. We have therefore identified a mechanism for LPS signaling which involves a decrease in miR-107 leading to an increase in CDK6. PMID- 21628466 TI - Bone morphogenetic protein 3 controls insulin gene expression and is down regulated in INS-1 cells inducibly expressing a hepatocyte nuclear factor 1A maturity-onset diabetes of the young mutation. AB - Inactivating mutations in the transcription factor hepatocyte nuclear factor (HNF) 1A cause HNF1A-maturity-onset diabetes of the young (HNF1A-MODY), the most common monogenic form of diabetes. To examine HNF1A-MODY-induced defects in gene expression, we performed a microarray analysis of the transcriptome of rat INS-1 cells inducibly expressing the common hot spot HNF1A frameshift mutation, Pro291fsinsC-HNF1A. Real-time quantitative PCR (qPCR), Western blotting, immunohistochemistry, reporter assays, and chromatin immunoprecipitation (ChIP) were used to validate alterations in gene expression and to explore biological activities of target genes. Twenty-four hours after induction of the mutant HNF1A protein, we identified a prominent down-regulation of the bone morphogenetic protein 3 gene (Bmp-3) mRNA expression. Reporter assays, qPCR, and Western blot analysis validated these results. In contrast, inducible expression of wild-type HNF1A led to a time-dependent increase in Bmp-3 mRNA and protein levels. Moreover, reduced protein levels of BMP-3 and insulin were detected in islets of transgenic HNF1A-MODY mice. Interestingly, treatment of naive INS-1 cells or murine organotypic islet cultures with recombinant human BMP-3 potently increased their insulin levels and restored the decrease in SMAD2 phosphorylation and insulin gene expression induced by the HNF1A frameshift mutation. Our study suggests a critical link between HNF1A-MODY-induced alterations in Bmp-3 expression and insulin gene levels in INS-1 cells and indicates that the reduced expression of growth factors involved in tissue differentiation may play an important role in the pathophysiology of HNF1A-MODY. PMID- 21628467 TI - Transit defect of potassium-chloride Co-transporter 3 is a major pathogenic mechanism in hereditary motor and sensory neuropathy with agenesis of the corpus callosum. AB - Missense and protein-truncating mutations of the human potassium-chloride co transporter 3 gene (KCC3) cause hereditary motor and sensory neuropathy with agenesis of the corpus callosum (HMSN/ACC), which is a severe neurodegenerative disease characterized by axonal dysfunction and neurodevelopmental defects. We previously reported that KCC3-truncating mutations disrupt brain-type creatine kinase-dependent activation of the co-transporter through the loss of its last 140 amino acids. Here, we report a novel and more distal HMSN/ACC-truncating mutation (3402C -> T; R1134X) that eliminates only the last 17 residues of the protein. This small truncation disrupts the interaction with brain-type creatine kinase in mammalian cells but also affects plasma membrane localization of the mutant transporter. Although it is not truncated, the previously reported HMSN/ACC-causing 619C -> T (R207C) missense mutation also leads to KCC3 loss of function in Xenopus oocyte flux assay. Immunodetection in Xenopus oocytes and in mammalian cultured cells revealed a decreased amount of R207C at the plasma membrane, with significant retention of the mutant proteins in the endoplasmic reticulum. In mammalian cells, curcumin partially corrected these mutant protein mislocalizations, with more protein reaching the plasma membrane. These findings suggest that mis-trafficking of mutant protein is an important pathophysiological feature of HMSN/ACC causative KCC3 mutations. PMID- 21628468 TI - A major portion of DNA gyrase inhibitor microcin B17 undergoes an N,O-peptidyl shift during synthesis. AB - Microcin B17 (McB) is a 43-amino acid antibacterial peptide targeting the DNA gyrase. The McB precursor is ribosomally produced and then post-translationally modified by the McbBCD synthase. Active mature McB contains eight oxazole and thiazole heterocycles. Here, we show that a major portion of mature McB contains an additional unusual modification, a backbone ester bond connecting McB residues 51 and 52. The modification results from an N -> O shift of the Ser(52) residue located immediately downstream of one of McB thiazole heterocycles. We speculate that the N,O-peptidyl shift undergone by Ser(52) is an intermediate of post translational modification reactions catalyzed by the McbBCD synthase that normally lead to formation of McB heterocycles. PMID- 21628469 TI - Side chain-oxidized oxysterols regulate the brain renin-angiotensin system through a liver X receptor-dependent mechanism. AB - Disturbances in cholesterol metabolism have been associated with hypertension and neurodegenerative disorders. Because cholesterol metabolism in the brain is efficiently separated from plasma cholesterol by the blood-brain barrier (BBB), it is an unsolved paradox how high blood cholesterol can cause an effect in the brain. Here, we discuss the possibility that cholesterol metabolites permeable to the BBB might account for these effects. We show that 27-hydroxycholesterol (27 OH) and 24S-hydroxycholesterol (24S-OH) up-regulate the renin-angiotensin system (RAS) in the brain. Brains of mice on a cholesterol-enriched diet showed up regulated angiotensin converting enzyme (ACE), angiotensinogen (AGT), and increased JAK/STAT activity. These effects were confirmed in in vitro studies with primary neurons and astrocytes exposed to 27-OH or 24S-OH, and were partially mediated by liver X receptors. In contrast, brain RAS activity was decreased in Cyp27a1-deficient mice, a model exhibiting reduced 27-OH production from cholesterol. Moreover, in humans, normocholesterolemic patients with elevated 27-OH levels, due to a CYP7B1 mutation, had markers of activated RAS in their cerebrospinal fluid. Our results demonstrate that side chain-oxidized oxysterols are modulators of brain RAS. Considering that levels of cholesterol and 27-OH correlate in the circulation and 27-OH can pass the BBB into the brain, we suggest that this cholesterol metabolite could be a link between high plasma cholesterol levels, hypertension, and neurodegeneration. PMID- 21628470 TI - Hepatitis C virus NS5B protein delays s phase progression in human hepatocyte derived cells by relocalizing cyclin-dependent kinase 2-interacting protein (CINP). AB - Cell cycle dysregulation is a critical event in virus infection-associated tumorigenesis. Previous studies have suggested that hepatitis C virus NS5B modulates cell cycle progression in addition to participating in RNA synthesis as an RNA-dependent RNA polymerase. However, the molecular mechanisms have thus far remained unclear. In this study, a HepG2 Tet-On NS5B stable cell line was generated to confirm the effect of NS5B on the cell cycle. To better understand the role of NS5B in cell cycle regulation, yeast two-hybrid assays were performed using a human liver cDNA library. The cyclin-dependent kinase 2-interacting protein (CINP) was identified. The interaction between NS5B and CINP was further demonstrated by in vivo and in vitro assays, and their association was found to be indispensable for S phase delay and cell proliferation suppression. Further experiments indicated that NS5B relocalized CINP from the nucleus to the cytoplasm. Directly knocking down CINP by specific siRNA resulted in a significant alteration in the DNA damage response and expression of cell cycle checkpoint proteins, including an increase in p21 and a decrease in phosphorylated Retinoblastoma and Chk1. Similar results were observed in cells expressing NS5B, and the effects were partially reversed upon ectopic overexpression of CINP. These studies suggest that the DNA damage response might be exploited by NS5B to hinder cell cycle progression. Taken together, our data demonstrate that NS5B delays cells in S phase through interaction with CINP and relocalization of the protein from the nucleus to the cytoplasm. Such effects might contribute to hepatitis C virus persistence and pathogenesis. PMID- 21628471 TI - Glycosylation regulates specific induction of rice immune responses by Acidovorax avenae flagellin. AB - Plants have a sensitive system that detects various pathogen-derived molecules to protect against infection. Flagellin, a main component of the bacterial flagellum, from the rice avirulent N1141 strain of the Gram-negative phytopathogenic bacterium Acidovorax avenae induces plant immune responses including H2O generation, whereas flagellin from the rice virulent K1 strain of A. avenae does not induce these immune responses. To clarify the molecular mechanism that leads to these differing responses between the K1 and N1141 flagellins, recombinant K1 and N1141 flagellins were generated using an Escherichia coli expression system. When cultured rice cells were treated with recombinant K1 or N1141 flagellin, both flagellins equally induced H2O2 generation, suggesting that post-translational modifications of the flagellins are involved in the specific induction of immune responses. Mass spectrometry analyses using glycosyltransferase-deficient mutants showed that 1,600- and 2,150 Da glycans were present on the flagellins from N1141 and K1, respectively. A deglycosylated K1 flagellin induced immune responses in the same manner as N1141 flagellin. Site-directed mutagenesis revealed that glycans were attached to four amino acid residues (Ser178, Ser183, Ser212, and Thr351) in K1 flagellin. Among mutant K1 flagellins in which each glycan-attached amino acid residue was changed to alanine, S178A and S183A, K1 flagellin induced a strong immune response in cultured rice cells, indicating that the glycans at Ser178 and Ser183 in K1 flagellin prevent epitope recognition in rice. PMID- 21628477 TI - Treatment of cutaneous sporotrichosis with itraconazole--study of 645 patients. AB - BACKGROUND: Itraconazole has become the first choice for treatment of cutaneous sporotrichosis. However, this recommendation is based on case reports and small series. The safety and efficacy of itraconazole were evaluated in 645 patients who received a diagnosis on the basis of isolation of Sporothrix schenckii in Rio de Janeiro, Brazil. METHODS: A standard regimen of itraconazole (100 mg/day orally) was used. Clinical and laboratory adverse events were assessed a grades 1 4. A multivariate Cox model was used to analyze the response to treatment. RESULTS: The median age was 43 years. Lymphocutaneous form occurred in 68.1% and fixed form in 23.1%. Six hundred ten patients (94.6%) were cured with itraconazole (50-400 mg/day): 547 with 100 mg/day, 59 with 200-400 mg/day, and 4 children with 50 mg/day. Three patients switched to potassium iodide, 2 to terbinafine, and 4 to thermotherapy. Twenty-six were lost to follow-up. Clinical adverse events occurred in 18.1% of patients using 100 mg/day and 21.9% of those using 200-400 mg/day. The most frequent clinical adverse events were nausea and epigastric pain. Laboratory adverse events occurred in 24.1%; the most common was hypercholesterolemia, followed by hypertriglyceridemia. Four hundred sixty-two patients (71.6%) completed clinical follow-up, and all remained cured. Only 2 variables were significant in explaining the cure: patients with erythema nodosum healed faster, and lymphocutaneous form took longer to cure. CONCLUSIONS: In the current series, the therapeutic response was excellent with the minimum dose of itraconazole, and there was a low incidence of adverse events and treatment failure. PMID- 21628472 TI - Neural cell adhesion molecule potentiates the growth of murine melanoma via beta catenin signaling by association with fibroblast growth factor receptor and glycogen synthase kinase-3beta. AB - The neural cell adhesion molecule (NCAM) was recently shown to be involved in the progression of various tumors with diverse effects. We previously demonstrated that NCAM potentiates the cellular invasion and metastasis of melanoma. Here we further report that the growth of melanoma is obviously retarded when the expression of NCAM is silenced. We found that the proliferation of murine B16F0 melanoma cells, their colony formation on soft agar, and growth of transplanted melanoma in vivo are clearly inhibited by the introduction of NCAM siRNA. Interestingly, change of NCAM expression level is shown to regulate the activity of Wnt signaling molecule, beta-catenin, markedly. This novel machinery requires the function of FGF receptor and glycogen synthase kinase-3beta but is independent of the Wnt receptors, MAPK-Erk and PI3K/Akt pathways. In addition, NCAM is found to form a functional complex with beta-catenin, FGF receptor, and glycogen synthase kinase-3beta. Moreover, up-regulation of NCAM140 and NCAM180 appears more potent than NCAM120 in activation of beta-catenin, suggesting that the intracellular domain of NCAM is required for facilitating the beta-catenin signaling. Furthermore, the melanoma cells also exhibit distinct differentiation phenotypes with the NCAM silencing. Our findings reveal a novel regulatory role of NCAM in the progression of melanoma that might serve as a new therapeutic target for the treatment of melanoma. PMID- 21628479 TI - Transmission elasticity in communities hyperendemic for tuberculosis. AB - BACKGROUND: Despite consistently meeting international performance targets for tuberculosis case detection and treatment success, areas where tuberculosis is hyperendemic fail to achieve the predicted epidemiological impact. In this article, we explore the anomalous relationship between defined performance targets and actual reduction in tuberculosis transmission. METHODS: In areas where tuberculosis is endemic, poorly ventilated social gathering places such as shebeens (informal alcohol drinking places), minibus taxis, and clinic waiting rooms are all potential transmission hot spots. We modeled the transmission reduction achieved by removal of infectious persons in settings with different tuberculosis prevalence rates to demonstrate the concept of transmission elasticity. We then applied this concept to real-life data from a hyperendemic community in Cape Town, South Africa. RESULTS: In a hyperendemic area, reducing the number of infectious people by a given percentage results in a smaller percentage decrease in the annual risk of infection (ARI) compared with a nonendemic area; for example, removing 10% of infectious persons could result in as little as a 5% reduction in the ARI. With use of real-life data and removal of 60% of infectious individuals with tuberculosis, as would be achieved by meeting current performance targets of 70% case detection and 85% cure, the estimated ARI reduction is 50%. CONCLUSIONS: The relationship between the number of infectious people removed and the decrease in ARI is nonlinear. The concept of transmission elasticity has important implications for the formulation of universal performance targets, since hyperendemic areas would require more stringent targets to achieve comparable transmission reduction. PMID- 21628480 TI - High titers of pre-existing adenovirus serotype-specific neutralizing antibodies in the host predict viral reactivation after allogeneic stem cell transplantation in children. AB - BACKGROUND: Human adenovirus (HAdV) infections are frequent in children after allogeneic stem cell transplantation (SCT) and may become fatal. Whether these infections occur through reactivation of endogenous virus or transmission via the graft remains a matter of debate. METHODS: In a cohort of 24 pediatric patients who received SCT, infections with 1 or more of 5 serotypes of HAdV (1, 2, 5, 6, and 31) were detected by culture. Neutralizing antibody (NAb) titers were measured in vitro by means of a virus neutralization assay. RESULTS: In 11 patients the infection was restricted to 1 site as demonstrated by culture only, and in 13 patients the HAdV infection was disseminated because plasma samples contained HAdV DNA. The 5 most commonly encountered HAdV serotypes caused 35 infectious episodes after SCT. Serum titers of NAb against these 5 serotypes of HAdV were measured before and after transplantation. High titers of NAb against a certain serotype in the recipient prior to SCT, reflecting previous infection, appeared to predispose for infection with the same serotype after SCT. In only 1 case of 41 independent samples of graft material, a very low level of HAdV DNA was detected. Antibody responses after SCT were detected in 21 of 35 infectious episodes. CONCLUSIONS: Together, these data suggest that adenoviral complications after SCT are caused by reactivation of endogenous persistent HAdV rather than by de novo infection from the donor or environment. This finding may offer a strategy of prophylactic treatment of high-risk patients before SCT to prevent infectious complications after allogeneic SCT. PMID- 21628481 TI - Sex differences in the incidence and case fatality rates from hemorrhagic fever with renal syndrome in China, 2004-2008. AB - BACKGROUND: Differences between male and female individuals in response to infectious diseases are an overlooked global health problem. METHODS: The relationship between sex and disease outcome was examined in populations of patients with hemorrhagic fever with renal syndrome (HFRS) in mainland China, where most cases of hantavirus exposure occur. HFRS in China is diagnosed on the basis of symptoms and is confirmed with serological testing. The geographical distribution, incidence, and case fatality rates (CFRs) of HFRS in China were estimated and compared by patient sex and age. In a subset of patients with HFRS, clinical manifestations of HFRS were assessed using latent class analysis and compared by sex. RESULTS: There were 80,671 HFRS cases reported during the period 2004-2008, with a majority of HFRS cases (39.2%) occurring among individuals 20 39 years of age. The incidence of HFRS was higher among male patients than among female patients for all individuals >10 years of age. There were 945 deaths (CFR, 1.17%) due to HFRS in China during the period 2004-2008. CFRs were higher among women than among men between the ages of 20-39 and >= 50 years of age. There were no sex differences in the geographical distribution of HFRS cases or deaths. Although the prevalence of each clinical marker did not differ by sex, 2 profiles of clinical markers were identified that were related to both severity of disease and sex. CONCLUSIONS: These data illustrate a paradox in which the incidence of disease is greater for males, but the severity of disease outcome is worse for females. Several behavioral, societal, and biological factors are hypothesized to be involved. PMID- 21628482 TI - Impact of combined low-level mupirocin and genotypic chlorhexidine resistance on persistent methicillin-resistant Staphylococcus aureus carriage after decolonization therapy: a case-control study. AB - BACKGROUND: The clinical importance of low-level mupirocin resistance and genotypic chlorhexidine resistance remains unclear. We aimed to determine whether resistance to these agents increases the risk of persistent methicillin-resistant Staphylococcus aureus (MRSA) carriage after their use for topical decolonization therapy. METHODS: A nested case-control study was conducted of MRSA carriers who received decolonization therapy from 2001 through 2008. Cases, patients who remained colonized, were matched by year to controls, those in whom MRSA was eradicated (follow-up, 2 years). Baseline MRSA isolates were tested for mupirocin resistance by Etest and chlorhexidine resistance by qacA/B polymerase chain reaction. MRSA carriers with high-level mupirocin resistance were excluded. The effect of the primary exposure of interest, low-level mupirocin and genotypic chlorhexidine resistance, was evaluated with multivariate conditional logistic regression analysis. RESULTS: The 75 case patients and 75 control patients were similar except that those persistently colonized were older (P = .007) with longer lengths of hospital stay (P = .001). After multivariate analysis, carriage of combined low-level mupirocin and genotypic chlorhexidine resistance before decolonization independently predicted persistent MRSA carriage (odds ratio [OR], 3.4 [95% confidence interval {CI}, 1.5-7.8]). Other risk factors were older age (OR, 1.04 [95% CI, 1.02-1.1]), previous hospitalization (OR, 2.4 [95% CI, 1.1 5.7]), presence of a skin wound (OR, 5.7 [95% CI, 1.8-17.6]), recent antibiotic use (OR, 3.1 [95% CI, 1.3-7.2]), and central venous catheterization (OR, 5.7 [95% CI, 1.4-23.9]). CONCLUSIONS: Combined low-level mupirocin and genotypic chlorhexidine resistance significantly increases the risk of persistent MRSA carriage after decolonization therapy. Institutions with widespread use of these agents should monitor for resistance and loss of clinical effectiveness. PMID- 21628483 TI - Follow-up of 686 patients with acute Q fever and detection of chronic infection. AB - BACKGROUND: Recent outbreaks in the Netherlands allowed for laboratory follow-up of a large series of patients with acute Q fever and for evaluation of test algorithms to detect chronic Q fever, a condition with considerable morbidity and mortality. METHODS: For 686 patients with acute Q fever, IgG antibodies to Coxiella burnetii were determined using an immunofluorescence assay at 3, 6, and 12 months of follow-up. Polymerase chain reaction (PCR) was performed after 12 months and on earlier serum samples with an IgG phase I antibody titer >= 1:1024. RESULTS: In 43% of patients, the IgG phase II antibody titers remained high (>= 1:1024) at 3, 6, and 12 months of follow-up. Three months after acute Q fever, 14% of the patients had an IgG phase I titer >= 1:1024, which became negative later in 81%. IgG phase I antibody titers were rarely higher than phase II titers. Eleven cases of chronic Q fever were identified on the basis of serological profile, PCR results, and clinical presentation. Six of these patients were known to have clinical risk factors at the time of acute Q fever. In a comparison of various serological algorithms, IgG phase I titer >= 1:1024 at 6 months had the most favorable sensitivity and positive predictive value for the detection of chronic Q fever. CONCLUSIONS: The wide variation of serological and PCR results during the follow-up of acute Q fever implies that the diagnosis of chronic Q fever, necessitating long-term antibiotic treatment, must be based primarily on clinical grounds. Different serological follow-up strategies are needed for patients with and without known risk factors for chronic Q fever. PMID- 21628484 TI - Microbiology of bronchoalveolar lavage fluid in children with acute nonresponding or recurrent community-acquired pneumonia: identification of nontypeable Haemophilus influenzae as a major pathogen. AB - BACKGROUND: Precise etiologic diagnosis in pediatric community-acquired pneumonia (CAP) remains challenging. METHODS: We conducted a retrospective study of CAP etiology in 2 groups of pediatric patients who underwent flexible bronchoscopy (FOB) with bronchoalveolar lavage (BAL); children with acute nonresponsive CAP (NR-CAP; n = 127) or recurrent CAP (Rec-CAP; n = 123). Procedural measures were taken to limit contamination risk and quantitative bacterial culture of BAL fluid (significance cutoff point, >= 104 colony-forming units/mL) was used. Blood culture results, serological test results, nasopharyngeal secretion findings, and pleural fluid culture results were also assessed, where available. RESULTS: An infectious agent was detected in 76.0% of cases. In 51.2% of infections, aerobic bacteria were isolated, of which 75.0%, 28.9%, and 13.3% were Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae, respectively. Most (97.9%) of the H. influenzae strains were nontypeable (NTHi). H. influenzae was detected in 26.0% of NR-CAP cases and 51.2% of Rec-CAP cases, whereas Mycoplasma pneumoniae was the predominant pathogen in the NR-CAP group (accounting for 34.9% of cases) but not in the Rec-CAP group (19.3%). Viruses were found in 30.4% of cases, with respiratory syncytial virus, parainfluenzaviruses, and influenzaviruses detected most frequently. Mixed infections were found in 18.9% of NR-CAP cases and 30.1% of Rec-CAP cases. CONCLUSIONS: A variety of microorganisms were isolated with frequent mixed infection. NTHi was one of the major pathogens found, especially in association with recurrent CAP, possibly because of improved detection with the FOB with BAL procedure. This suggests that the burden of pediatric CAP could be reduced by addressing NTHi as a major causative pathogen. PMID- 21628485 TI - A 24-year-old barber with proptosis and seizures. PMID- 21628486 TI - CLABSI rates in immunocompromised patients: a valuable patient centered outcome? AB - The accepted approach to surveillance for hospital-acquired bloodstream infection (HABSI) due to central venous catheters requires use of the National Health and Safety Network (NHSN) definition for catheter-associated bloodstream infection (CLABSI). In this commentary, we discuss our experience with the application of current NHSN surveillance definitions for CLABSI and the impact that public reporting of CLABSI rates in settings with a high prevalence of special populations has on infection prevention (IP) programs. For IP programs to serve the continuous improvement needs of their organizations, surveillance methodologies need to accurately capture the burden of preventable HABSI among immunocompromised individuals with inherent risk for infection. Current NHSN CLABSI definitions lack specificity for complex and heterogeneous patient populations and require modification. Beyond definitions, IP programs must critically assess the value of their current approach to surveillance to assure that patient-centered outcomes are the focus of prevention efforts. PMID- 21628487 TI - Effective utilization of evolving methods for the laboratory diagnosis of Clostridium difficile infection. AB - Physicians should understand the performance characteristics of evolving laboratory tests used to diagnose Clostridium difficile infection if they are to correctly integrate test results with clinical information and formulate an appropriate therapeutic intervention for patients with antibiotic-associated diarrhea. PMID- 21628488 TI - Immunodeficiency as a risk factor for non-AIDS-defining malignancies in HIV-1 infected patients receiving combination antiretroviral therapy. AB - BACKGROUND: The aim of this study was to investigate the association between immunodeficiency, viremia, and non-AIDS-defining malignancies (NADM). METHODS: Patients starting combination antiretroviral therapy (cART) as of 1 January 1996 were selected from the AIDS Therapy Evaluation in the Netherlands (ATHENA) cohort. In Cox models, risk factors for NADM were investigated. These included age, sex, transmission route, smoking, alcohol abuse, prior AIDS diagnosis, duration of exposure to cART, and estimated duration of human immunodeficiency virus infection. CD4+ cell count and viral load (VL) were considered as time updated variables and as measures of cumulative exposure to CD4+ cell counts of < 200, < 350, or < 500 cells/mm3 and detectable VL >50, >400, and >1000 copies/mL, respectively. RESULTS: In a cohort of 11,459 patients, 236 NADMs were diagnosed; 102 were caused by infection, and 134 were attributable to other causes. Median CD4+ cell count at NADM diagnosis was 340 cells/mm3 (range, 210-540 cells/mm3). Median time to first NADM after starting cART was 5.0 years (range, 2.2-8.2 years). In multivariate models, cumulative exposure to CD4+ cell counts < 200 cells/mm3 remained significant (hazard ratio [HR], 1.12; range, 1.03-1.22) for each additional year of exposure. In stratified analyses, cumulative exposure to CD4+ cell counts < 200 cells/mm3 was associated with malignancies possibly caused by infection (HR, 1.16; range, 1.03-1.31]) but was not associated with other types of cancers. No significant effect of viremia was seen in either type of cancer. CONCLUSIONS: Cumulative exposure to CD4+ cell counts < 200 cells/mm3 during cART was associated with an increased risk of infection-related non-AIDS defining malignancies. PMID- 21628490 TI - Treatment of skin and soft tissue infections due to community-associated methicillin-resistant Staphylococcus aureus in Europe: the role of trimethoprim sulfamethoxazole0010. PMID- 21628491 TI - A closer look at aerosolized colistin. PMID- 21628492 TI - In vivo programming of tumor antigen-specific T lymphocytes from pluripotent stem cells to promote cancer immunosurveillance. AB - Adoptive T-cell immunotherapy has garnered wide attention, but its effective use is limited by the need of multiple ex vivo manipulations and infusions that are complex and expensive. In this study, we show how highly reactive antigen (Ag) specific CTLs can be generated from induced pluripotent stem (iPS) cells to provide an unlimited source of functional CTLs for adoptive immunotherapy. iPS cell-derived T cells can offer the advantages of avoiding possible immune rejection and circumventing ethical and practical issues associated with other stem cell types. iPS cells can be differentiated into progenitor T cells in vitro by stimulation with the Notch ligand Delta-like 1 (DL1) overexpressed on bone marrow stromal cells, with complete maturation occurring upon adoptive transfer into Rag1-deficient mice. Here, we report that these iPS cells can be differentiated in vivo into functional CTLs after overexpression of MHC I restricted Ag-specific T-cell receptors (TCR). In this study, we generated murine iPS cells genetically modified with ovalbumin (OVA)-specific and MHC-I restricted TCR (OT-I) by retrovirus-mediated transduction. After their adoptive transfer into recipient mice, the majority of OT-I/iPS cells underwent differentiation into CD8+ CTLs. TCR-transduced iPS cells developed in vivo responded in vitro to peptide stimulation by secreting interleukin 2 and IFN-gamma. Most importantly, adoptive transfer of TCR-transduced iPS cells triggered infiltration of OVA reactive CTLs into tumor tissues and protected animals from tumor challenge. Taken together, our findings offer proof of concept for a potentially more efficient approach to generate Ag-specific T lymphocytes for adoptive immunotherapy. PMID- 21628493 TI - Heterogeneity maintenance in glioblastoma: a social network. AB - Glioblastoma multiforme (GBM), the most common intracranial tumor in adults, is characterized by extensive heterogeneity at the cellular and molecular levels. This insidious feature arises inevitably in almost all cancers and has great significance for the general outcome of the malignancy, because it confounds our understanding of the disease and also intrinsically contributes to the tumor's aggressiveness and poses an obstacle to the design of effective therapies. The classic view that heterogeneity arises as the result of a tumor's "genetic chaos" and the more contemporary cancer stem cell (CSC) hypothesis tend to identify a single cell population as the therapeutic target: the prevailing clone over time in the first case and the CSC in the latter. However, there is growing evidence that the different tumor cell populations may not be simple bystanders. Rather, they can establish a complex network of interactions between each other and with the tumor microenvironment that eventually strengthens tumor growth and increases chances to escape therapy. These differing but complementary ideas about the origin and maintenance of tumor heterogeneity and its importance in GBM are reviewed here. PMID- 21628495 TI - Speech stimulation during functional MR imaging as a potential indicator of autism. AB - PURPOSE: To determine the feasibility of applying functional magnetic resonance (MR) imaging as an objective indicator of language disability in autism by using passive speech stimulation. MATERIALS AND METHODS: This prospective study was approved by the institutional review board, and informed consent was obtained from the parents or guardians of all subjects. Functional MR imaging was performed during passive presentations of prerecorded speech in 15 control subjects (mean age +/- standard deviation, 12.1 years +/- 4.3) and 12 language impaired, age-matched autistic subjects (mean age, 12.4 years +/- 4.7). An additional 27 autistic children (mean age, 8.4 years +/- 3.1), who underwent imaging while sedated with propofol as part of routine clinical MR evaluations, were also included. Activation maps for each subject were computed by using univariate general linear model analyses. The spread (quantified as number of voxels) and amplitude of the functional MR imaging activation were then quantified within two anatomically specified regions of interest known to be involved with language: the primary auditory cortex (A1) and the superior temporal gyrus (STG). Group differences were compared by using analysis of variance, two-sample t tests, and Wilcoxon rank sum tests where appropriate. The threshold for autism was defined as 1 standard deviation below the control mean for subjects imaged in the alert state. A similar threshold was estimated for sedated autistic subjects on the basis of differences between nonsedated and sedated autistic subjects. RESULTS: Activity in A1 did not differ between autistic and control subjects. However, mean amplitude and spread of activity in the STG differed between autistic and control subjects (P < .001). Values for 10 of the 12 (83%) nonsedated autistic subjects decreased at least 1 standard deviation below the control distribution. The threshold derived from sedation adjusted values of the control group enabled identification of 26 of the 27 (96%) sedated autistic subjects. CONCLUSION: Functional MR imaging activation within the STG in response to passive speech stimulation helped differentiate autistic from control subjects, demonstrating the potential utility of functional MR imaging as an objective indicator of language impairment in autism. Future studies may lead to an early and objective indicator for autism with these methods. PMID- 21628494 TI - Detection of promoter hypermethylation in salivary rinses as a biomarker for head and neck squamous cell carcinoma surveillance. AB - PURPOSE: Hypermethylation of tumor suppressor gene promoters has been found in head and neck squamous carcinoma (HNSCC) and other solid tumors. We evaluated these alterations in pretreatment salivary rinses from HNSCC patients by using real-time quantitative methylation-specific PCR (Q-MSP). EXPERIMENTAL DESIGN: Pretreatment saliva DNA samples from HNSCC patients were evaluated for patterns of hypermethylation by using Q-MSP. Target tumor suppressor gene promoter regions were selected based on a previous study describing a screening panel for HNSCC in a high-risk population subjects. The selected genes were: DAPK, DCC, MINT-31, TIMP-3, p16, MGMT, CCNA1. RESULTS: We analyzed the panel in a cohort of 61 HNSCC patients. Thirty-three of the analyzed patients (54.1%) showed methylation of at least one of the selected genes in the saliva DNA. Pretreatment methylated saliva DNA was not significantly associated with tumor site (P = 0.209) nor clinical stage (P = 0.299). However, local disease control and overall survival were significantly lower in patients presenting hypermethylation in saliva rinses (P = 0.010 and P = 0.015, respectively). Multivariate analysis confirmed that this hypermethylation pattern remained as an independent prognostic factor for local recurrence (HR = 12.2; 95% CI = 1.8-80.6; P = 0.010) and overall survival (HR = 2.8; 95% CI = 1.2-6.5; P = 0.016). CONCLUSIONS: We were able to confirm an elevated rate of promoter hypermethylation in HNSCC saliva of patients by using a panel of gene promoters previously described as methylated specifically in HNSCC. Detection of hypermethylation in pretreatment saliva DNA seems to be predictive of local recurrence and overall survival. This finding has potential to influence treatment and surveillance of HNSCC patients. PMID- 21628496 TI - pH-Dependent transport of pemetrexed by breast cancer resistance protein. AB - Breast cancer resistance protein (BCRP), an ATP-dependent efflux transporter, confers drug resistance to many chemotherapy agents. BCRP is overexpressed in tumors exposed to an acidic environment; therefore, it is important to establish the effect of low pH on BCRP transport activity. It has recently been reported that BCRP transports substrates more efficiently in an acidic microenvironment. In the study presented here, we examine the pH dependence of BCRP using methothrexate (MTX), pemetrexed (PMX), and estrone sulfate (ES) as model substrates. Our study revealed an increase of approximately 40-fold in the BCRP mediated transport of PMX and MTX when the pH was decreased from 7.4 to 5.5. In contrast, only a 2-fold increase was observed for ES. These results indicate a mechanism of transport that is directly dependent on the effective ionization state of the substrates and BCRP. For ES, which retains a constant ionization state throughout the applied pH, the observed mild increase in activity is attributable to the overall changes in the effective ionization state and conformation of BCRP. For MTX and PMX, the marked increase in BCRP transport activity was likely due to the change in ionization state of MTX and PMX at lowered pH and their intermolecular interactions with BCRP. To further rationalize the molecular basis of the pH dependence, molecular modeling and docking studies were carried out using a homology model of BCRP, which has previously been closely examined in structural and site-directed mutagenesis studies (Am J Physiol Cell Physiol 299:C1100-C1109, 2010). On the basis of docking studies, all model compounds were found to associate with arginine 482 (Arg482) by direct salt-bridge interactions via their negatively charged carboxylate or sulfate groups. However, at lower pH, protonated MTX and PMX formed an additional salt-bridge interaction between their positively charged moieties and the nearby negatively charged aspartic acid 477 (Asp477) carboxylate side chain. The formation of this "salt-bridge triad" is expected to increase the overall electrostatic interactions between MTX and PMX with BCRP, which can form a rational basis for the pH dependence of the observed enhanced binding selectivity and transport activity. Removal of Arg482 in site-directed mutagenesis studies eliminated this pH dependence, which lends further support to our binding model. These results shed light on the importance of electrostatic interactions in transport activity and may have important implications in the design of ionizable chemotherapeutics intended for tumors in the acidic microenvironment. PMID- 21628497 TI - Bioactivation of glafenine by human liver microsomes and peroxidases: identification of electrophilic iminoquinone species and GSH conjugates. AB - Glafenine (Privadol; 2,3-dihydroxypropyl 2-[(7-chloro-4-quinolinyl) amino]benzoate) is a non-narcotic analgesic agent widely used for the treatment of pains of various origins. Severe liver toxicity and a high incidence of anaphylaxis were reported in patients treated with glafenine, eventually leading to its withdrawal from the market in most countries. It is proposed that bioactivation of glafenine and subsequent binding of reactive metabolite(s) to critical cellular proteins play a causative role. The study described herein aimed at characterizing pathways of glafenine bioactivation and the metabolic enzymes involved. Two GSH conjugates of glafenine were detected in human liver microsomal incubations using liquid chromatography tandem mass spectrometry. The structures of detected conjugates were determined as GSH adducts of 5 hydroxyglafenine (M3) and 5-hydroxy glafenic acid (M4), respectively. GSH conjugation took place with a strong preference at C6 of the benzene ring of glafenine, ortho to the carbonyl moiety. These findings are consistent with a bioactivation sequence involving initial cytochrome P450-catalyzed 5 hydroxylation of the benzene ring of glafenine, followed by two electron oxidations of M3 and M4 to form corresponding para-quinone imine intermediates that react with GSH to form GSH adducts M1 and M2, respectively. Formation of M1 and M2 was primarily catalyzed by heterologously expressed recombinant CYP3A4 and to a lesser extent, CYP2C19 and CYP2D6. We demonstrated that M3 can also be bioactivated by peroxidases, such as horseradish peroxidase and myeloperoxidase. In summary, these findings have significance in understanding the bioactivation pathways of glafenine and their potential link to mechanisms of toxicity of glafenine. PMID- 21628498 TI - Metabolism of [(14)C]GSK977779 in rats and its implication with the observed covalent binding. AB - GSK977779 is a potent HM74a agonist evaluated for the treatment of dyslipidemia. The disposition and metabolism of [(14)C]GSK977779 (67.6 MUmol/kg p.o.) was studied in male and female rats. The compound was well absorbed and its primary route of elimination was in the feces. Based on metabolite profiling of plasma extracts and urine and bile samples, it was demonstrated that GSK977779 was extensively metabolized in the rat by N-dealkylation, mono- and dioxygenation, reductive and oxidative cleavage of the 1,2,4-oxadiazole ring, and conjugative pathways. After plasma extraction high amounts of nonextractable radioactivity were observed, which were more pronounced in female rats. Size-exclusion chromatography and SDS gel electrophoresis indicated that the majority of the nonextractable radioactivity was covalently bound to plasma proteins. Solubilization of the plasma protein pellet followed by high-performance liquid chromatography and mass spectrometry suggested that a carboxylic acid metabolite derived from oxadiazole ring cleavage may be responsible for the observed covalent binding of the radioactivity to rat plasma proteins. PMID- 21628499 TI - Paracetamol-induced hypothermia is independent of cannabinoids and transient receptor potential vanilloid-1 and is not mediated by AM404. AB - In recent years, there has been increasing interest in hypothermia induced by paracetamol for therapeutic purposes, which, in some instances, has been reported as a side effect. Understanding the mechanism by which paracetamol induces hypothermia is therefore an important question. In this study, we investigated whether the novel metabolite of paracetamol, N-(4-hydroxyphenyl)arachidonylamide (AM404), which activates the cannabinoid (CB) and transient receptor potential vanilloid-1 (TRPV1) systems, mediates the paracetamol-induced hypothermia. The hypothermic response to 300 mg/kg paracetamol in CB(1) receptor (CB(1)R) and TRPV1 knockout mice was compared to wild-type mice. Hypothermia induced by paracetamol was also investigated in animals pretreated with the CB(1)R or TRPV1 antagonist 1-(2,4-dichlorophenyl)-5-(4-iodophenyl)-4-methyl-N-1-piperdinyl-1H pyrazole-3-carboxamide trifluoroacetate salt (AM251) or 4'-chloro-3 methoxycinnamanilide (SB366791), respectively. In CB(1)R or TRPV1 knockout mice, paracetamol induced hypothermia to the same extent as in wild-type mice. In addition, in C57BL/6 mice pretreated with AM251 or SB366791, paracetamol induced hypothermia to the same extent as in control mice. AM404 failed to induce hypothermia at pharmacological doses. Inhibition of fatty acid amide hydrolase (FAAH), which is involved in the metabolism of paracetamol to AM404, did not prevent the development of hypothermia with paracetamol. Paracetamol also induced hypothermia in FAAH knockout mice to the same extent as wild-type mice. We conclude that paracetamol induces hypothermia independent of cannabinoids and TRPV1 and that AM404 does not mediate this response. In addition, potential therapeutic value of combinational drug-induced hypothermia is supported by experimental evidence. PMID- 21628500 TI - Lack of association between the Tlr4 (Lpsd/Lpsd) genotype and increased susceptibility to Escherichia coli bladder infections in female C3H/HeJ mice. AB - Toll-like receptor 4 is thought to have a primary role in host defense against Escherichia coli bladder colonization, based on mouse models of urinary tract infection using C3H/HeJ female mice. This strain carries a point mutation in the Tlr4 gene, which renders the mice unresponsive to lipopolysaccharide (LPS) and thus limits the bladder inflammatory response and infection resolution. The importance of Tlr4 as the sole genetic determinant of resistance or susceptibility can be questioned, however, by the observation that C3H/HeOuJ female mice with a functional Tlr4 do not effectively resolve E. coli bladder infections. The present study further examined this inconsistency by investigating the association of Tlr4 Lps(d) and Lps(n) alleles with bladder infection susceptibility by using genetic crosses of C3H/HeJ mice with Tlr4 (Lps(n)/Lps(n)) or (Lps(n)/Lps(d)) mice. Heterozygous offspring of C3H/HeJ (Lps(d)/Lps(d)) * BALB/cAnN (Lps(n)/Lps(n)) mice successfully resolved bladder infections induced by a uropathogenic E. coli strain, while heterozygous mice from a C3H/HeJ (Lps(d)/Lps(d)) * C3H/HeOuJ (Lps(n)/Lps(n)) cross had severe infections. A backcross of C3H/HeJ (Lps(d)/Lps(d)) with (BALB/cAnN * C3H/HeJ)F(1) (Lps(n)/Lps(d)) produced mice that were either resistant or susceptible to E. coli bladder infections and had Lps(d)/Lps(d) or Lps(n)/Lps(d) Tlr4 genotypes. The Lps(d)/Lps(d) or Lps(n)/Lps(d) genotypes were present in individual mice with unresolved bladder infections, and the Lps(d)/Lps(d) genotype was found in infection-resistant mice. These results indicate that at least one gene other than Tlr4 strongly influences susceptibility to E. coli bladder infections in C3H/HeJ mice. IMPORTANCE: We have previously demonstrated that mouse strains with either a functional or nonfunctional Tlr4 were not able to resolve induced Escherichia coli bladder infections and that a chromosomal site distinct from Tlr4 was associated with an inability to resolve bladder infections in C3H/HeJ mice. The present study has further investigated the relevance of Tlr4 in bladder infection resolution by defining the Tlr4 alleles present in offspring of genetic crosses of C3H/HeJ mice with infection-resistant and -susceptible inbred strains. The results of these experiments showed that mice with a normal Tlr4 on different genetic backgrounds were not able to clear E. coli bladder infections and that animals with a defective Tlr4 could successfully resolve infections. These results strongly imply the presence of a gene other than in Tlr4 as an important genetic determinant of infection resistance/susceptibility in C3H/HeJ and other inbred mouse strains used in mouse models of infectious diseases. PMID- 21628501 TI - An unusual mechanism of isopeptide bond formation attaches the collagenlike glycoprotein BclA to the exosporium of Bacillus anthracis. AB - The outermost exosporium layer of spores of Bacillus anthracis, the causative agent of anthrax, is comprised of a basal layer and an external hairlike nap. The nap includes filaments composed of trimers of the collagenlike glycoprotein BclA. Essentially all BclA trimers are tightly attached to the spore in a process requiring the basal layer protein BxpB (also called ExsFA). Both BclA and BxpB are incorporated into stable, high-molecular-mass complexes, suggesting that BclA is attached directly to BxpB. The 38-residue amino-terminal domain of BclA, which is normally proteolytically cleaved between residues 19 and 20, is necessary and sufficient for basal layer attachment. In this study, we demonstrate that BclA attachment occurs through the formation of isopeptide bonds between the free amino group of BclA residue A20 and a side chain carboxyl group of an acidic residue of BxpB. Ten of the 13 acidic residues of BxpB can participate in isopeptide bond formation, and at least three BclA polypeptide chains can be attached to a single molecule of BxpB. We also demonstrate that similar cross linking occurs in vitro between purified recombinant BclA and BxpB, indicating that the reaction is spontaneous. The mechanism of BclA attachment, specifically, the formation of a reactive amino group by proteolytic cleavage and the promiscuous selection of side chain carboxyl groups of internal acidic residues, appears to be different from other known mechanisms for protein cross-linking through isopeptide bonds. Analogous mechanisms appear to be involved in the cross linking of other spore proteins and could be found in unrelated organisms. IMPORTANCE: Isopeptide bonds are protein modifications found throughout nature in which amide linkages are formed between functional groups of two amino acids, with at least one of the functional groups provided by an amino acid side chain. Isopeptide bonds generate cross-links within and between proteins that are necessary for proper protein structure and function. In this study, we discovered that BclA, the dominant structural protein of the external nap of Bacillus anthracis spores, is attached to the underlying exosporium basal layer protein BxpB via isopeptide bonds formed through a mechanism fundamentally different from previously described mechanisms of isopeptide bond formation. The most unusual features of this mechanism are the generation of a reactive amino group by proteolytic cleavage and promiscuous selection of acidic side chains. This mechanism, which apparently relies only on short peptide sequences in protein substrates, could be a general mechanism in vivo and adapted for protein cross linking in vitro. PMID- 21628502 TI - Surviving bacterial sibling rivalry: inducible and reversible phenotypic switching in Paenibacillus dendritiformis. AB - Natural habitats vary in available nutrients and room for bacteria to grow, but successful colonization can lead to overcrowding and stress. Here we show that competing sibling colonies of Paenibacillus dendritiformis bacteria survive overcrowding by switching between two distinct vegetative phenotypes, motile rods and immotile cocci. Growing colonies of the rod-shaped bacteria produce a toxic protein, Slf, which kills cells of encroaching sibling colonies. However, sublethal concentrations of Slf induce some of the rods to switch to Slf resistant cocci, which have distinct metabolic and resistance profiles, including resistance to cell wall antibiotics. Unlike dormant spores of P. dendritiformis, the cocci replicate. If cocci encounter conditions that favor rods, they secrete a signaling molecule that induces a switch to rods. Thus, in contrast to persister cells, P. dendritiformis bacteria adapt to changing environmental conditions by inducible and reversible phenotypic switching. IMPORTANCE: In favorable environments, species may face space and nutrient limits due to overcrowding. Bacteria provide an excellent model for analyzing principles underlying overcrowding and regulation of density in nature, since their population dynamics can be easily and accurately assessed under controlled conditions. We describe a newly discovered mechanism for survival of a bacterial population during overcrowding. When competing with sibling colonies, Paenibacillus dendritiformis produces a lethal protein (Slf) that kills cells at the interface of encroaching colonies. Slf also induces a small proportion of the cells to switch from motile, rod-shaped cells to nonmotile, Slf-resistant, vegetative cocci. When crowding is reduced and nutrients are no longer limiting, the bacteria produce a signal that induces cocci to switch back to motile rods, allowing the population to spread. Genes encoding components of this phenotypic switching pathway are widespread among bacterial species, suggesting that this survival mechanism is not unique to P. dendritiformis. PMID- 21628504 TI - Potential transcriptional regulatory regions exist upstream of the human ezrin gene promoter in esophageal carcinoma cells. AB - We previously demonstrated that the region -87/+134 of the human ezrin gene (VIL2) exhibited promoter activity in human esophageal carcinoma EC109 cells, and a further upstream region -1324/-890 positively regulated transcription. In this study, to identify the transcriptional regulatory regions upstream of the VIL2 promoter, we cloned VIL2 -1541/-706 segment containing the -1324/-890, and investigated its transcriptional regulatory properties via luciferase assays in transiently transfected cells. In EC109 cells, it was found that VIL2 -1541/-706 possessed promoter and enhancer activities. We also localized transcriptional regulatory regions by fusing 5'- or 3'-deletion segments of VIL2 -1541/-706 to a luciferase reporter. We found that there were three positive and one negative transcriptional regulatory regions within VIL2 -1541/-706 in EC109 cells. When these regions were separately located upstream of the luciferase gene without promoter, or located upstream of the VIL2 promoter or SV40 promoter directing the luciferase gene, only VIL2 -1297/-1186 exhibited considerable promoter and enhancer activities, which were lower than those of -1541/-706. In addition, transient expression of Sp1 increased ezrin expression and the transcriptional activation of VIL2 -1297/-1186. Other three regions, although exhibiting significantly positive or negative transcriptional regulation in deletion experiments, showed a weaker or absent regulation. These data suggested that more than one region upstream of the VIL2 promoter participated in VIL2 transcription, and the VIL2 -1297/-1186, probably as a key transcriptional regulatory region, regulated VIL2 transcription in company with other potential regulatory regions. PMID- 21628503 TI - Surface export of GAPDH/SDH, a glycolytic enzyme, is essential for Streptococcus pyogenes virulence. AB - Streptococcal surface dehydrogenase (SDH) (glyceraldehyde-3-phosphate dehydrogenase [GAPDH]) is an anchorless major multifunctional surface protein in group A Streptococcus (GAS) with the ability to bind important mammalian proteins, including plasmin(ogen). Although several biological properties of SDH are suggestive of its possible role in GAS virulence, its direct role in GAS pathogenesis has not been ascertained because it is essential for GAS survival. Thus, it has remained enigmatic as to "how and why" SDH/GAPDH is exported onto the bacterial surface. The present investigation highlights "why" SDH is exported onto the GAS surface. Differential microarray-based genome-wide transcript abundance analysis was carried out using a specific mutant, which was created by inserting a hydrophobic tail at the C-terminal end of SDH (M1-SDH(HBtail)) and thus preventing its exportation onto the GAS surface. This analysis revealed downregulation of the majority of genes involved in GAS virulence and genes belonging to carbohydrate and amino acid metabolism and upregulation of those related to lipid metabolism. The complete attenuation of this mutant for virulence in the mouse model and the decreased and increased virulence of the wild-type and mutant strains postcomplementation with SDH(HBtail) and SDH, respectively, indicated that the SDH surface export indeed regulates GAS virulence. M1-SDH(HBtail) also displayed unaltered growth patterns, increased intracellular ATP concentration and Hpr double phosphorylation, and significantly reduced pH tolerance, streptolysin S, and SpeB activities. These phenotypic and physiological changes observed in the mutant despite the unaltered expression levels of established transcriptional regulators further highlight the fact that SDH interfaces with many regulators and its surface exportation is essential for GAS virulence. IMPORTANCE: Streptococcal surface dehydrogenase (SDH), a classical anchorless cytoplasmically localized glycolytic enzyme, is exported onto the group A Streptococcus (GAS) surface through a hitherto unknown mechanism(s). It has not been known why GAS or other prokaryotes should export this protein onto the surface. By genetic manipulations, we created a novel GAS mutant strain expressing SDH with a 12-amino-acid hydrophobic tail at its C-terminal end and thus were able to prevent its surface exportation without altering its enzymatic activity or growth pattern. Interestingly, the mutant was completely attenuated for virulence in a mouse peritonitis model. The global gene expression profiles of this mutant reveal that the surface exportation of SDH is mandatory to maintain GAS virulence. The ability of GAS as a successful pathogen to localize SDH in the cytoplasm as well as on the surface is physiologically relevant and dynamically obligatory to fine-tune the functions of many transcriptional regulators and also to exploit its virulence properties for infection. PMID- 21628505 TI - Histone deacetylase inhibitor valproic acid sensitizes B16F10 melanoma cells to cucurbitacin B treatment. AB - Cucurbitacin B (CuB) is reported to have anti-proliferation effects on a variety of tumors including melanoma, and more effective regimens by combination of this agent with others are under investigation. In this study, the anti-melanoma effect of CuB as a single agent and in combination with valproic acid (VPA), an inhibitor of histone deacetylase (HDAC), was evaluated in B16F10, a mouse melanoma cell line. The results demonstrated that CuB inhibited the proliferation of the cell line in a dose-dependent manner. However, it was likely that a pro survival compensatory response, involving the induction of autophagy and upregulation of anti-apoptotic Bcl-2 protein, was induced by CuB treatment, which might greatly decrease the cytotoxicity of this agent. Supporting this, the melanoma cells were found to be more sensitive to the combination of CuB with chloroquine, a well-known autophagy inhibitor. And CuB-induced autophagy was associated with c-Jun N-terminal kinase (JNK) activation, at least partly, since inhibition of JNK activity by SP600125 could alleviate the autophagy. When CuB was combined with VPA, the two drugs showed synergistic cytotoxicity by induction of cell apoptosis. Moreover, the multiploidization effect of CuB was also suppressed in the presence of VPA. In contrast to the transient activation of JNKs by CuB, the combination of CuB and VPA resulted in prolonged JNK activation, although at low level after 4 h. Our results demonstrated that HDAC inhibitor VPA can sensitize B16F10 cells to CuB treatment through induction of apoptotic pathway. PMID- 21628510 TI - TLR4 gene polymorphisms: evidence for protection against type 2 diabetes but not for diabetes-associated ischaemic heart disease. AB - OBJECTIVE: Several factors either predisposing or protecting from the onset of diabetes mellitus type 2 (DM2) have been proposed. Two specific polymorphisms of toll-like receptor 4 (TLR4; Asp299Gly and Thr399Ile) have recently been identified either as candidate protector genes against DM2 and associated neuropathy or risk alleles for the manifestation of diabetic retinopathy. The impact of these alleles on the risk for ischaemic heart disease (IHD) is controversial while their role in diabetes-associated IHD has never been studied. DESIGN AND METHODS: In order to clarify the potential impact of TLR4 polymorphisms on the predisposition for DM2 as well as on diabetes-related IHD vulnerability, the distribution of the mutant TLR4 Asp299Gly and Thr399Ile alleles in 286 DM2 patients and 413 non-DM2 controls with or without IHD, was examined. RESULTS: Mutant alleles were predominantly detected in 79/413 non diabetic individuals versus 15/286 DM2 patients (P<0.0001). The rates of positivity for mutant alleles were similar among diabetic patients with or without IHD (7/142 vs 8/144, P>0.1), whereas they proved different among non diabetic individuals with or without IHD (39/145 vs 40/268, P=0.004). Following multivariate analysis, the difference between diabetic and non-diabetic subjects, with regard to TLR4 mutations alone, remained significant (P=0.04). CONCLUSIONS: Mutant TLR4 alleles confer protection against DM2. However, their presence does not seem to play any role, protective or aggravating, in the manifestation of IHD either in diabetic or in non-diabetic individuals. PMID- 21628511 TI - Sterile protective immunity to malaria is associated with a panel of novel P. falciparum antigens. AB - The development of an effective malaria vaccine remains a global public health priority. Less than 0.5% of the Plasmodium falciparum genome has been assessed as potential vaccine targets and candidate vaccines have been based almost exclusively on single antigens. It is possible that the failure to develop a malaria vaccine despite decades of effort might be attributed to this historic focus. To advance malaria vaccine development, we have fabricated protein microarrays representing 23% of the entire P. falciparum proteome and have probed these arrays with plasma from subjects with sterile protection or no protection after experimental immunization with radiation attenuated P. falciparum sporozoites. A panel of 19 pre-erythrocytic stage antigens was identified as strongly associated with sporozoite-induced protective immunity; 16 of these antigens were novel and 85% have been independently identified in sporozoite and/or liver stage proteomic or transcriptomic data sets. Reactivity to any individual antigen did not correlate with protection but there was a highly significant difference in the cumulative signal intensity between protected and not protected individuals. Functional annotation indicates that most of these signature proteins are involved in cell cycle/DNA processing and protein synthesis. In addition, 21 novel blood-stage specific antigens were identified. Our data provide the first evidence that sterile protective immunity against malaria is directed against a panel of novel P. falciparum antigens rather than one antigen in isolation. These results have important implications for vaccine development, suggesting that an efficacious malaria vaccine should be multivalent and targeted at a select panel of key antigens, many of which have not been previously characterized. PMID- 21628512 TI - Development and application of a mouse intestinal loop model to study the in vivo action of Clostridium perfringens enterotoxin. AB - Clostridium perfringens enterotoxin (CPE) is responsible for causing the gastrointestinal symptoms of C. perfringens type A food poisoning, the second most commonly identified bacterial food-borne illness in the United States. CPE is produced by sporulating C. perfringens cells in the small intestinal lumen, where it then causes epithelial cell damage and villous blunting that leads to diarrhea and cramping. Those effects are typically self-limiting; however, severe outbreaks of this food poisoning, particularly two occurring in psychiatric institutions, have involved deaths. Since animal models are currently limited for the study of the CPE action, a mouse ligated intestinal loop model was developed. With this model, significant lethality was observed after 2 h in loops receiving an inoculum of 100 or 200 MUg of CPE but not using a 50-MUg toxin inoculum. A correlation was noted between the overall intestinal histological damage and lethality in mice. Serum analysis revealed a dose-dependent increase in serum CPE and potassium levels. CPE binding to the liver and kidney was detected, along with elevated levels of potassium in the serum. These data suggest that CPE can be absorbed from the intestine into the circulation, followed by the binding of the toxin to internal organs to induce potassium leakage, which can cause death. Finally, CPE pore complexes similar to those formed in tissue culture cells were detected in the intestine and liver, suggesting that (i) CPE actions are similar in vivo and in vitro and (ii) CPE-induced potassium release into blood may result from CPE pore formation in internal organs such as the liver. PMID- 21628513 TI - Differences in erythrocyte receptor specificity of different parts of the Plasmodium falciparum reticulocyte binding protein homologue 2a. AB - The Plasmodium falciparum reticulocyte-binding-like protein homologue (RH) and erythrocyte binding-like (EBL) protein families play important roles during invasion, though their exact roles are not clear. Both EBL and RH proteins are thought to directly bind different receptors on the surface of the erythrocyte, and the binding properties for a number of EBLs and RHs have been described. While P. falciparum RH1 (PfRH1) and PfRH4 have been shown to act directly in two alternative invasion pathways used by merozoites, the functions of PfRH2a and PfRH2b during invasion are less defined. Here, using monoclonal antibodies raised against a unique region of PfRH2a, we show that PfRH2a moves from the rhoptry neck to the moving junction during merozoite invasion. The movement of PfRH2a to the junction is independent of the invasion pathway used by the merozoite, suggesting an additional function of the protein that is independent of receptor binding. We further show that PfRH2a is processed both in the schizont and during invasion, resulting in proteins with different erythrocyte binding properties. Our findings suggest that PfRH2a and, most likely, the other members of the RH family, depending on their processing stage, can engage different receptors at different stages of the invasion process. PMID- 21628514 TI - PrsW is required for colonization, resistance to antimicrobial peptides, and expression of extracytoplasmic function sigma factors in Clostridium difficile. AB - Clostridium difficile is an anaerobic, Gram-positive, spore-forming, opportunistic pathogen that is the most common cause of hospital-acquired infectious diarrhea. In numerous pathogens, stress response mechanisms are required for survival within the host. Extracytoplasmic function (ECF) sigma factors are a major family of signal transduction systems, which sense and respond to extracellular stresses. We have identified three C. difficile ECF sigma factors. These ECF sigma factors, CsfT, CsfU, and CsfV, induce their own expressions and are negatively regulated by their cognate anti-sigma factors, RsiT, RsiU, and RsiV, respectively. The levels of expression of these ECF sigma factors increase following exposure to the antimicrobial peptides bacitracin and/or lysozyme. The expressions of many ECF sigma factors are controlled by site 1 and site 2 proteases, which cleave anti-sigma factors. Using a retargeted group II intron, we generated a C. difficile mutation in prsW, a putative site 1 protease. The C. difficile prsW mutant exhibited decreased levels of expression of CsfT and CsfU but not of CsfV. When expressed in a heterologous host, C. difficile PrsW was able to induce the degradation of RsiT but not of RsiU. When the prsW mutant was tested in competition assays against its isogenic parent in the hamster model of C. difficile infection, we found that the prsW mutant was 30 fold less virulent than the wild type. The prsW mutant was also significantly more sensitive to bacitracin and lysozyme than the wild type in in vitro competition assays. Taken together, these data suggest that PrsW likely regulates the activation of the ECF sigma factor CsfT in C. difficile and controls the resistance of C. difficile to antimicrobial peptides that are important for survival in the host. PMID- 21628515 TI - Enhanced egress of intracellular Eimeria tenella sporozoites by splenic lymphocytes from coccidian-infected chickens. AB - Egress, which describes the mechanism that some intracellular parasites use to exit from parasitophorous vacuoles and host cells, plays a very important role in the parasite life cycle and is central to Eimeria propagation and pathogenesis. Despite the importance of egress in the intracellular parasite's life cycle, very little information is known on this process compared to other steps, e.g., invasion. The present study was conducted to investigate the interplay between the host adaptive immune system and Eimeria egression. Splenic lymphocytes or soluble immune factors were incubated with parasite-infected host cells for 3 or 5 h, and the percentage of egress was calculated according to an established formula. Viability of egressed parasites and host cells was tested using trypan blue exclusion and annexin V and propidium iodide staining, respectively. We found that premature egression of sporozoites from Eimeria tenella-infected primary chicken kidney cells or from chicken peripheral blood mononuclear cells occurred when the cells were cocultured in vitro with spleen lymphocytes from E. tenella-infected chickens but not when they were cocultured with splenocytes from uninfected chickens. Eimeria-specific antibodies and cytokines (gamma interferon [IFN-gamma], interleukin-2 [IL-2], and IL-15), derived from E. tenella-primed B and T lymphocytes, respectively, were capable of promoting premature egress of sporozoites from infected host cells. Both egressed parasites and host cells were viable, although the latter showed reduced reinvasion ability. These results suggest a novel, immune-mediated mechanism that the host exploits to interrupt the normal Eimeria life cycle in vivo and thereby block the release of mature parasites into the environment. PMID- 21628516 TI - Oral exposure to Trypanosoma cruzi elicits a systemic CD8+ T cell response and protection against heterotopic challenge. AB - Trypanosoma cruzi infects millions of people in Latin America and often leads to the development of Chagas disease. T. cruzi infection can be acquired at or near the bite site of the triatomine vector, but per os infection is also a well documented mode of transmission, as evidenced by recent microepidemics of acute Chagas disease attributed to the consumption of parasite-contaminated foods and liquids. It would also be convenient to deliver vaccines for T. cruzi by the oral route, particularly live parasite vaccines intended for the immunization of reservoir hosts. For these reasons, we were interested in better understanding immunity to T. cruzi following oral infection or oral vaccination, knowing that the route of infection and site of antigen encounter can have substantial effects on the ensuing immune response. Here, we show that the route of infection does not alter the ability of T. cruzi to establish infection in muscle tissue nor does it impair the generation of a robust CD8(+) T cell response. Importantly, oral vaccination with attenuated parasites provides protection against wild-type (WT) T. cruzi challenge. These results strongly support the development of whole organism-based vaccines targeting reservoir species as a means to alleviate the burden of Chagas disease in affected regions. PMID- 21628517 TI - Antibodies in action: role of human opsonins in killing Salmonella enterica serovar Typhi. AB - Although vaccines have been available for over a century, a correlate of protection for typhoid fever has yet to be identified. Antibodies are produced in response to typhoid infection and vaccination and are generally used as the gold standard for determining vaccine immunogenicity, even though their role in clearance of Salmonella enterica serovar Typhi infections is poorly defined. Here, we describe the first functional characterization of S. Typhi-specific antibodies following vaccination with a new vaccine, M01ZH09 (Ty2 DeltaaroC DeltassaV). We determined that postvaccination sera increased the uptake of wild type S. Typhi by human macrophages up to 2.3-fold relative to prevaccination (day 0) or placebo samples. These results were recapitulated using immunoglobulins purified from postvaccination serum, demonstrating that antibodies were largely responsible for increases in uptake. Imaging verified that macrophages internalized 2- to 9.5-fold more S. Typhi when the bacteria were opsonized with postvaccination sera than when the bacteria were opsonized with day 0 or placebo sera. Once inside macrophages, the survival of S. Typhi was reduced as much as 50% when opsonized with postvaccination sera relative to day 0 or placebo serum samples. Lastly, bactericidal assays indicated that antibodies generated postvaccination were recognized by complement factors and assisted in killing S. Typhi: mean postvaccination bactericidal antibody titers were higher at all time points than placebo and day 0 titers. These data clearly demonstrate that there are at least two mechanisms by which antibodies facilitate killing of S. Typhi. Future work could lead to improved immunogenicity tests associated with vaccine efficacy and the identification of correlates of protection against typhoid fever. PMID- 21628518 TI - Multigenic control and sex bias in host susceptibility to spore-induced pulmonary anthrax in mice. AB - Mechanisms underlying susceptibility to anthrax infection are unknown. Using a phylogenetically diverse panel of inbred mice and spores of Bacillus anthracis Ames, we investigated host susceptibility to pulmonary anthrax. Susceptibility profiles for survival time and organ pathogen load differed across strains, indicating distinct genetic controls. Tissue infection kinetics analysis showed greater systemic dissemination in susceptible DBA/2J (D) mice but a higher terminal bacterial load in resistant BALB/cJ (C) mice. Interestingly, the most resistant strains, C and C57BL/6J (B), demonstrated a sex bias for susceptibility. For example, BALB/cJ females had a significantly higher survival time and required 4-fold more spores for 100% mortality compared to BALB/cJ males. To identify genetic regions associated with differential susceptibility, survival time and extent of organ infection were assessed using mice derived from two susceptibility models: (i) BXD advanced recombinant inbred strains and (ii) F2 offspring generated from polar responding C and D strains. Genome-wide analysis of BXD strain survival identified linkage on chromosomes 5, 6, 9, 11, and 14. Quantitative trait locus (QTL) analysis of the C*DF2 population revealed a significant QTL (designated Rpai1 for resistance to pulmonary anthrax infection, locus 1) for survival time on chromosome 17 and also identified a chromosome 11 locus for lung pathogen burden. The striking difference between genome-wide linkage profiles for these two mouse models of anthrax susceptibility supports our hypothesis that these are multigenic traits. Our data provide the first evidence for a differential sex response to anthrax resistance and further highlight the unlikelihood of a single common genetic contribution for this response across strains. PMID- 21628519 TI - CD1d-independent activation of invariant natural killer T cells by staphylococcal enterotoxin B through major histocompatibility complex class II/T cell receptor interaction results in acute lung injury. AB - There are two important mechanisms of activation of invariant natural killer T cells (iNKT cells) by microbes: direct activation of the invariant T-cell receptor (TCR) by microbial glycolipids presented by CD1d and indirect activation, mediated by the responses of antigen-presenting cells to microbes. In this study, we provide evidence for a novel CD1d-independent direct activation of iNKT cells involving a microbial protein superantigen presented in the context of major histocompatibility complex class II (MHC-II), which plays a critical role in pathogenesis, thereby redefining the role of iNKT cells. Intranasal exposure to staphylococcal enterotoxin B (SEB) in C57BL/6 wild-type mice caused acute lung injury (ALI) characterized by vascular leak, cytokine storm, and infiltration of mononuclear cells in the lungs. In contrast, the vascular leak and inflammation were decreased by ~50% in NKT cell-deficient Jalpha18(-/-) and CD1d(-/-) mice following SEB exposure, which was reversed following adoptive transfer of iNKT cells into CD1d(-/-) mice. In vitro, SEB could directly stimulate iNKT cells in a CD1d-independent manner via MHC-II/TCR interaction, specifically involving Vbeta8. These studies not only demonstrate that iNKT cells can be activated directly by a bacterial protein superantigen independent of CD1d but also indicate that in addition to the conventional T cells, iNKT cells play a critical role in SEB-mediated ALI. PMID- 21628520 TI - Expression of Toll-like receptor 4 and downstream effectors in selected cecal cell subpopulations of chicks resistant or susceptible to Salmonella carrier state. AB - Toll-like receptor 4 (TLR4), which recognizes lipopolysaccharide from Gram negative bacteria, plays a major role in resistance of mice and humans to Salmonella infection. In chickens, Salmonella may establish a carrier state whereby bacteria are able to persist in the host organism without triggering clinical signs. Based on cellular morphological parameters, we developed a method, without using antibodies, to separate three cecal cell subpopulations: lymphocytes, enterocytes, and a population encompassing multiple cell types. We analyzed the mRNA expression of TLR4, interleukin-1beta (IL-1beta), IL-8, IL-12, and lipopolysaccharide-induced tumor necrosis factor alpha factor (LITAF) in cecal subpopulations of chicks from inbred lines resistant or susceptible to the carrier state infected with Salmonella enterica serovar Enteritidis. The results showed that resistance to the carrier state in chicks is associated with a larger percentage of lymphocytes and with higher levels of expression of TLR4 and IL-8 at homeostasis in the three cell subpopulations, as well as with a higher level of expression of LITAF in lymphocytes during the carrier state. In contrast to the early phase of infection, the carrier state is characterized by no major cell recruitment differences between infected and noninfected animals and no significant modification in terms of TLR4, IL-1beta, IL-8, IL-12, and LITAF expression in all cell subpopulations measured. However, TLR4 expression increased in the lymphocytes of chicks from the susceptible line, reaching the same level as that in infected chicks from the resistant line. These observations suggest that the carrier state is characterized by a lack of immune activation and highlight the interest of working at the level of the cell population rather than that of the organ. PMID- 21628521 TI - Efficacy of a conjugate vaccine containing polymannuronic acid and flagellin against experimental Pseudomonas aeruginosa lung infection in mice. AB - Vaccines that could effectively prevent Pseudomonas aeruginosa pulmonary infections in the settings of cystic fibrosis (CF) and nosocomial pneumonia could be exceedingly useful, but to date no effective immunotherapy targeting this pathogen has been successfully developed for routine use in humans. Evaluations using animals and limited human trials of vaccines and their associated immune effectors against different P. aeruginosa antigens have suggested that antibody to the conserved surface polysaccharide alginate, as well as the flagellar proteins, often give high levels of protection. However, alginate itself does not elicit protective antibody in humans, and flagellar vaccines containing the two predominant serotypes of this antigen may not provide sufficient coverage against variant flagellar types. To evaluate if combining these antigens in a conjugate vaccine would be potentially efficacious, we conjugated polymannuronic acid (PMA), containing the blocks of mannuronic acid conserved in all P. aeruginosa alginates, to type a flagellin (FLA) and evaluated immunogenicity, opsonic killing activity, and passive protective efficacy in mice. The PMA-FLA conjugate was highly immunogenic in mice and rabbits and elicited opsonic antibodies against mucoid but not nonmucoid P. aeruginosa, but nonetheless rabbit antibody to PMA-FLA showed evidence of protective efficacy against both types of this organism in a mouse lung infection model. Importantly, the PMA-FLA conjugate vaccine did not elicit antibodies that neutralized the Toll-like receptor 5 (TLR5)-activating activity of flagellin, an important part of innate immunity to flagellated microbial pathogens. Conjugation of PMA to FLA appears to be a promising path for developing a broadly protective vaccine against P. aeruginosa. PMID- 21628522 TI - Migratory activation of primary cortical microglia upon infection with Toxoplasma gondii. AB - Disseminated toxoplasmosis in the central nervous system (CNS) is often accompanied by a lethal outcome. Studies with murine models of infection have focused on the role of systemic immunity in control of toxoplasmic encephalitis, while knowledge remains limited on the contributions of resident cells with immune functions in the CNS. In this study, the role of glial cells was addressed in the setting of recrudescent Toxoplasma infection in mice. Activated astrocytes and microglia were observed in the close vicinity of foci with replicating parasites in situ in the brain parenchyma. Toxoplasma gondii tachyzoites were allowed to infect primary microglia and astrocytes in vitro. Microglia were permissive to parasite replication, and infected microglia readily transmigrated across transwell membranes and cell monolayers. Thus, infected microglia, but not astrocytes, exhibited a hypermotility phenotype reminiscent of that recently described for infected dendritic cells. In contrast to gamma interferon-activated microglia, Toxoplasma-infected microglia did not upregulate major histocompatibility complex (MHC) class II molecules and the costimulatory molecule CD86. Yet Toxoplasma-infected microglia and astrocytes exhibited increased sensitivity to T cell-mediated killing, leading to rapid parasite transfer to effector T cells in vitro. We hypothesize that glial cells and T cells, besides their role in triggering antiparasite immunity, may also act as "Trojan horses," paradoxically facilitating dissemination of Toxoplasma within the CNS. To our knowledge, this constitutes the first report of migratory activation of a resident CNS cell by an intracellular parasite. PMID- 21628523 TI - Oligopeptide permease A5 modulates vertebrate host-specific adaptation of Borrelia burgdorferi. AB - Borrelia burgdorferi, the agent of Lyme disease, undergoes rapid adaptive gene expression in response to signals unique to its arthropod vector or vertebrate hosts. Among the upregulated genes under vertebrate host conditions is one of the five annotated homologs of oligopeptide permease A (OppA5, BBA34). A mutant lacking oppA5 was constructed in an lp25-deficient isolate of B. burgdorferi strain B31, and the minimal regions of infectivity were restored via a shuttle vector pBBE22 with or without an intact copy of bba34. Immunoblot analysis of the bba34 mutant revealed a reduction in the levels of RpoS, BosR, and CsrA(Bb) with a concomitant reduction in the levels of OspC, DbpA, BBK32, and BBA64. There were no changes in the levels of OspA, NapA, P66, and three other OppA orthologs. Quantitative transcriptional analysis correlated with the changes in the protein levels. However, the bba34 mutant displayed comparable infectivities in the C3H/HeN mice and the wild-type strain, despite the reduction in several pathogenesis-related proteins. Supplementation of the growth medium with increased levels of select components, notably sodium acetate and sodium bicarbonate, restored the levels of several proteins in the bba34 mutant to wild type levels. We speculate that the transport of acetate appears to contribute to the accumulation of key metabolites, like acetyl phosphate, that facilitate the adaptation of B. burgdorferi to the vertebrate host by the activation of the Rrp2 RpoN-RpoS pathway. These studies underscore the importance of solute transport to host-specific adaptation of B. burgdorferi. PMID- 21628524 TI - CXCR6 is a marker for protective antigen-specific cells in the lungs after intranasal immunization against Mycobacterium tuberculosis. AB - Convincing correlates of protective immunity against tuberculosis have been elusive. In BALB/c mice, intranasal immunization with a replication-deficient recombinant adenovirus expressing Mycobacterium tuberculosis antigen 85A (adenovirus-85A) induces protective lower respiratory tract immunity against pulmonary challenge with Mycobacterium tuberculosis, while intradermal immunization with adenovirus-85A does not. Here we report that intranasal immunization with adenovirus-85A induces expression of the chemokine receptor CXCR6 on lung CD8 T lymphocytes, which is maintained for at least 3 months. CXCR6 positive antigen-specific T cell numbers are increased among bronchoalveolar lavage-recoverable cells. Similarly, intranasal immunization with recombinant antigen 85A with adjuvant induces CXCR6 expression on lung CD4 cells in BALB/c and C57BL/6 mice, while a synthetic ESAT6(1-20) peptide with adjuvant induces CXCR6 expression in C57BL/6 mice. Parenteral immunization fails to do so. Upregulation of CXCR6 is accompanied by a transient elevation of serum CXCL16 after intranasal immunization, and lung cells cultured ex vivo from mice immunized intranasally show increased production of CXCL16. Administration of CXCL16 and cognate antigen intranasally to mice previously immunized parenterally increases the number of antigen-specific T lymphocytes in the bronchoalveolar lavage-recoverable population, which mediates inhibition of the early growth of Mycobacterium tuberculosis after challenge. We conclude that expression of CXCR6 on lung T lymphocytes is a correlate of local protective immunity against Mycobacterium tuberculosis after intranasal immunization and that CXCR6 and CXCL16 play an important role in the localization of T cells within lung tissue and the bronchoalveolar lavage-recoverable compartment. PMID- 21628525 TI - A member of the PLEIOTROPIC DRUG RESISTANCE family of ATP binding cassette transporters is required for the formation of a functional cuticle in Arabidopsis. AB - Although the multilayered structure of the plant cuticle was discovered many years ago, the molecular basis of its formation and the functional relevance of the layers are not understood. Here, we present the permeable cuticle1 (pec1) mutant of Arabidopsis thaliana, which displays features associated with a highly permeable cuticle in several organs. In pec1 flowers, typical cutin monomers, such as omega-hydroxylated fatty acids and 10,16-dihydroxypalmitate, are reduced to 40% of wild-type levels and are accompanied by the appearance of lipidic inclusions within the epidermal cell. The cuticular layer of the cell wall, rather than the cuticle proper, is structurally altered in pec1 petals. Therefore, a significant role for the formation of the diffusion barrier in petals can be attributed to this layer. Thus, pec1 defines a new class of mutants. The phenotypes of the pec1 mutant are caused by the knockout of ATP BINDING CASSETTEG32 (ABCG32), an ABC transporter from the PLEIOTROPIC DRUG RESISTANCE family that is localized at the plasma membrane of epidermal cells in a polar manner toward the surface of the organs. Our results suggest that ABCG32 is involved in the formation of the cuticular layer of the cell wall, most likely by exporting particular cutin precursors from the epidermal cell. PMID- 21628526 TI - Ubp8 and SAGA regulate Snf1 AMP kinase activity. AB - Posttranslational modifications of histone proteins play important roles in the modulation of gene expression. The Saccharomyces cerevisiae (yeast) 2-MDa SAGA (Spt-Ada-Gcn5) complex, a well-studied multisubunit histone modifier, regulates gene expression through Gcn5-mediated histone acetylation and Ubp8-mediated histone deubiquitination. Using a proteomics approach, we determined that the SAGA complex also deubiquitinates nonhistone proteins, including Snf1, an AMP activated kinase. Ubp8-mediated deubiquitination of Snf1 affects the stability and phosphorylation state of Snf1, thereby affecting Snf1 kinase activity. Others have reported that Gal83 is phosphorylated by Snf1, and we found that deletion of UBP8 causes decreased phosphorylation of Gal83, which is consistent with the effects of Ubp8 loss on Snf1 kinase functions. Overall, our data indicate that SAGA modulates the posttranslational modifications of Snf1 in order to fine-tune gene expression levels. PMID- 21628527 TI - Selective ubiquitylation of p21 and Cdt1 by UBCH8 and UBE2G ubiquitin-conjugating enzymes via the CRL4Cdt2 ubiquitin ligase complex. AB - CRL4(Cdt2) is a cullin-based E3 ubiquitin ligase that promotes the ubiquitin dependent proteolysis of various substrates implicated in the control of cell cycle and various DNA metabolic processes such as DNA replication and repair. Substrates for CRL4(Cdt2) E3 ubiquitin ligase include the replication licensing factor Cdt1 and the cyclin-dependent kinase (Cdk) inhibitor p21. Inhibition of this E3 ligase leads to serious abnormalities of the cell cycle and cell death. The ubiquitin-conjugating enzyme (UBC) involved in this important pathway, however, remains unknown. By a proteomic analysis of Cdt2-associated proteins and an RNA interference-based screening approach, we show that CRL4(Cdt2) utilizes two different UBCs to target different substrates. UBCH8, a member of the UBE2E family of UBCs, ubiquitylates and promotes the degradation of p21, both during the normal cell cycle and in UV-irradiated cells. Importantly, depletion of UBCH8 by small interfering RNA (siRNA) increases p21 protein level, delays entry into S phase of the cell cycle, and suppresses the DNA damage response after UV irradiation. On the other hand, members of the UBE2G family of UBCs (UBE2G1 and UBE2G2) cooperate with CRL4(Cdt2) to polyubiquitylate and degrade Cdt1 postradiation, an activity that is critical for preventing origin licensing in DNA-damaged cells. Finally, we show that UBCH8, but not UBE2G1 or UBE2G2, is required for CRL4(Cdt2)-mediated ubiquitylation and degradation of the histone H4 lysine 20 monomethyltransferase Set8, a previously identified CRL4(Cdt2) substrate, as well as for CRL4(Cdt2)-dependent monoubiquitylation of PCNA in unstressed cells. These findings identify the UBCs required for the activity of CRL4(Cdt2) on multiple substrates and demonstrate that different UBCs are involved in the selective ubiquitylation of different substrates by the same E3 complex. PMID- 21628528 TI - Bub1 and BubR1: at the interface between chromosome attachment and the spindle checkpoint. AB - The spindle checkpoint ensures genome fidelity by temporarily halting chromosome segregation and the ensuing mitotic exit until the last kinetochore is productively attached to the mitotic spindle. At the interface between proper chromosome attachment and the metaphase-to-anaphase transition are the mammalian spindle checkpoint kinases. Compelling evidence indicates that the checkpoint kinases Bub1 and BubR1 have the added task of regulating kinetochore-microtubule attachments. However, the debate on the requirement of kinase activity is in full swing. This minireview summarizes recent advances in our understanding of the core spindle checkpoint kinases Bub1 and BubR1 and considers evidence that supports and opposes the role of kinase activity in regulating their functions during mitosis. PMID- 21628529 TI - Nonallelic transcriptional roles of CTCF and cohesins at imprinted loci. AB - The cohesin complex holds sister chromatids together and is essential for chromosome segregation. Recently, cohesins have been implicated in transcriptional regulation and insulation through genome-wide colocalization with the insulator protein CTCF, including involvement at the imprinted H19/Igf2 locus. CTCF binds to multiple imprinted loci and is required for proper imprinted expression at the H19/Igf2 locus. Here we report that cohesins colocalize with CTCF at two additional imprinted loci, the Dlk1-Dio3 and the Kcnq1/Kcnq1ot1 loci. Similar to the H19/Igf2 locus, CTCF and cohesins preferentially bind to the Gtl2 differentially methylated region (DMR) on the unmethylated maternal allele. To determine the functional importance of the binding of CTCF and cohesins at the three imprinted loci, CTCF and cohesins were depleted in mouse embryonic fibroblast cells. The monoallelic expression of imprinted genes at these three loci was maintained. However, mRNA levels for these genes were typically increased; for H19 and Igf2 the increased level of expression was independent of the CTCF-binding sites in the imprinting control region. Results of these experiments demonstrate an unappreciated role for CTCF and cohesins in the repression of imprinted genes in somatic cells. PMID- 21628530 TI - AMPK -> ULK1 -> autophagy. PMID- 21628531 TI - Inhibition of protein degradation induces apoptosis through a microtubule associated protein 1 light chain 3-mediated activation of caspase-8 at intracellular membranes. AB - The accumulation of damaged or misfolded proteins, if unresolved, can lead to a detrimental consequence within cells termed proteotoxicity. Since cancerous cells often display elevated protein synthesis and by-product disposal, inhibition of the protein degradation pathways is an emerging approach for cancer therapy. However, the molecular mechanism underlying proteotoxicity remains largely unclear. We show here that inhibition of proteasomal degradation results in an increased oligomerization and activation of caspase-8 on the cytosolic side of intracellular membranes. This enhanced caspase-8 oligomerization and activation are promoted through its interaction with the ubiquitin-binding protein SQSTM1/p62 and the microtubule-associated protein light chain 3 (LC3), which are enriched at intracellular membranes in response to proteotoxic stress. Silencing LC3 by shRNA, or the LC3 mutants defective in membrane localization or p62 interaction fail to induce caspase-8 activation and apoptosis. Our results unveiled a previously unknown mechanism through which disruption of protein homeostasis induces caspase-8 oligomerization, activation, and apoptosis. PMID- 21628532 TI - Topoisomerase 1 and single-strand break repair modulate transcription-induced CAG repeat contraction in human cells. AB - Expanded trinucleotide repeats are responsible for a number of neurodegenerative diseases, such as Huntington disease and myotonic dystrophy type 1. The mechanisms that underlie repeat instability in the germ line and in the somatic tissues of human patients are undefined. Using a selection assay based on contraction of CAG repeat tracts in human cells, we screened the Prestwick chemical library in a moderately high-throughput assay and identified 18 novel inducers of repeat contraction. A subset of these compounds targeted pathways involved in the management of DNA supercoiling associated with transcription. Further analyses using both small molecule inhibitors and small interfering RNA (siRNA)-mediated knockdowns demonstrated the involvement of topoisomerase 1 (TOP1), tyrosyl-DNA phosphodiesterase 1 (TDP1), and single-strand break repair (SSBR) in modulating transcription-dependent CAG repeat contractions. The TOP1 TDP1-SSBR pathway normally functions to suppress repeat instability, since interfering with it stimulated repeat contractions. We further showed that the increase in repeat contractions when the TOP1-TDP1-SSBR pathway is compromised arises via transcription-coupled nucleotide excision repair, a previously identified contributor to transcription-induced repeat instability. These studies broaden the scope of pathways involved in transcription-induced CAG repeat instability and begin to define their interrelationships. PMID- 21628534 TI - Single mutations in HIV integrase confer high-level resistance to raltegravir in primary human macrophages. AB - CD4(+) T cells and macrophages are the primary target cells for HIV in vivo, and antiretroviral drugs can vary in their ability to inhibit the infection of these different cell types. Resistance pathways to the HIV integrase inhibitor raltegravir have previously been investigated in T cells. Primary raltegravir resistance mutations, most often at integrase amino acid position 148 or 155, afford some resistance to the drug. The acquisition of pathway-specific secondary mutations then provides higher-level resistance to viruses infecting T cells. We show here that during macrophage infection, the presence of a single primary raltegravir resistance mutation (Q148H, Q148R, N155H, or N155S) is sufficient to provide resistance to raltegravir comparable to that seen in viruses expressing both primary and secondary mutations in costimulated CD4(+) T cells. These data implicate macrophages as a potential in vivo reservoir that may facilitate the development of resistance to raltegravir. Notably, the newer integrase inhibitor MK-2048 effectively suppressed the infection of all raltegravir-resistant viruses in both T cells and macrophages, indicating that more recently developed integrase inhibitors are capable of inhibiting infection in both major HIV cellular reservoirs, even in patients harboring raltegravir-resistant viruses. PMID- 21628535 TI - T cell monitoring of chemotherapy in experimental rat tuberculosis. AB - Mycobacterium tuberculosis is the causative agent of a pulmonary epidemic that is estimated to infect one-third of the world's population and that has an increased incidence of multidrug resistance. The evaluation of new chemical entities against M. tuberculosis is hampered by the lack of biological tools to help predict efficacy, from early drug development to clinical trials. As the rat is the animal species of choice in the pharmaceutical industry, we have developed a rat model of acute and chronic phases of M. tuberculosis infection for drug efficacy testing. In this model, we have evaluated the impact of tuberculosis drugs on T cell response using the enzyme-linked immunospot assay methodology. Infected rats treated with isoniazid (INH) or rifampin (RIF) responded to therapy, the potency of which was comparable to that seen in the mouse. Peripheral blood mononuclear cells from infected rats produced gamma interferon (IFN-gamma) in response to RD-1 antigens, such as the 6-kDa early secretory antigen target (ESAT-6) and the 10-kDa culture filtrate protein (CFP-10). A decrease in IFN-gamma spot-forming cells (SFCs) was consistently observed in response to drug treatment. In both the acute- and chronic-phase models, the T cell response was more sensitive to ESAT-6 than to CFP-10. The SFC count in response to ESAT-6 appears to be an indicator of bacterial killing in the rat. Collectively, our data suggest that the ESAT-6 response could be used as a potential surrogate of drug efficacy in the rat and that such a readout could help shorten drug testing during preclinical development. PMID- 21628536 TI - A novel cell-associated protection assay demonstrates the ability of certain antibiotics to protect ocular surface cell lines from subsequent clinical Staphylococcus aureus challenge. AB - In vivo effectiveness of topical antibiotics may depend on their ability to associate with epithelial cells to provide continued protection, but this contribution is not measured by standard antibiotic susceptibility tests. We report a new in vitro method that measures the ability of test antibiotics azithromycin (AZM), erythromycin (ERY), tetracycline (TET), and bacitracin (BAC) to associate with mammalian cells and to protect these cells from destruction by bacteria. Mammalian cell lines were grown to confluence using antibiotic-free medium and then incubated in medium containing a single antibiotic (0 to 512 MUg/ml). After incubation, the cells were challenged with Staphylococcus aureus ocular isolates, without antibiotics added to the culture medium. Epithelial cell layer integrity was assessed by gentian violet staining, and the minimum cell layer protective concentration (MCPC) of an antibiotic sufficient to protect the mammalian cells from S. aureus was determined. Staining was also quantified and analyzed. Bacterial viability was determined by culture turbidity and growth on agar plates. Preincubation of Chang and human corneal limbal epithelial cells with AZM, ERY, and TET at >=64 MUg/ml provided protection against AZM-susceptible S. aureus strains, with increasing protection at higher concentrations. TET toxicity was demonstrated at >64 MUg/ml, whereas AZM displayed toxicity to one cell line at 512 MUg/ml. BAC failed to show consistent protection at any dose, despite bacterial susceptibility to BAC as determined by traditional antibiotic susceptibility testing. A range of antibiotic effectiveness was displayed in this cell association assay, providing data that may be considered in addition to traditional testing when determining therapeutic dosing regimens. PMID- 21628537 TI - Candida glabrata mutants demonstrating paradoxical reduced caspofungin susceptibility but increased micafungin susceptibility. AB - Echinocandins, including caspofungin (CSP) and micafungin (MCF), are highly active versus Candida glabrata (MIC of <=0.06 MUg/ml). True resistance (MIC of >=1 MUg/ml) is a rare event and strictly associated with mutations in beta-1,3 glucan synthase gene FKS1 or FKS2. In contrast, we show here that mutants exhibiting reduced susceptibility to CSP (CRS; MICs of 0.12 to 0.5 MUg/ml) are readily selected in vitro and, paradoxically, demonstrate increased susceptibility to MCF (MIS) ranging from 4- to 32-fold. CRS-MIS mutants were generated from all 10 C. glabrata strains tested and were tentatively identified within a collection of clinical isolates. Intriguingly, sequencing and gene disruption demonstrated that CRS-MIS is Fks independent. PMID- 21628538 TI - Novel inhibitors of InhA efficiently kill Mycobacterium tuberculosis under aerobic and anaerobic conditions. AB - Drug resistance in Mycobacterium tuberculosis has become a serious global health threat, which is now complicated by the emergence of extensively drug-resistant strains. New drugs that are active against drug-resistant tuberculosis (TB) are needed. We chose to search for new inhibitors of the enoyl-acyl carrier protein (ACP) reductase InhA, the target of the first-line TB drug isoniazid (also known as isonicotinoic acid hydrazide [INH]). A subset of a chemical library, composed of 300 compounds inhibiting Plasmodium falciparum enoyl reductase, was tested against M. tuberculosis. Four compounds were found to inhibit M. tuberculosis growth with MICs ranging from 1 MUM to 10 MUM. Testing of these compounds against M. tuberculosis in vitro revealed that only two compounds (CD39 and CD117) were bactericidal against drug-susceptible and drug-resistant M. tuberculosis. These two compounds were also bactericidal against M. tuberculosis incubated under anaerobic conditions. Furthermore, CD39 and CD117 exhibited increased bactericidal activity when used in combination with INH or rifampin, but CD39 was shown to be toxic to eukaryotic cells. The compounds inhibit InhA as well the fatty acid synthase type I, and CD117 was found to also inhibit tuberculostearic acid synthesis. This study provides the TB drug development community with two chemical scaffolds that are suitable for structure-activity relationship study to improve on their cytotoxicities and bactericidal activities in vitro and in vivo. PMID- 21628539 TI - walK and clpP mutations confer reduced vancomycin susceptibility in Staphylococcus aureus. AB - Vancomycin-intermediate Staphylococcus aureus (VISA) is generated from vancomycin susceptible Staphylococcus aureus by multiple spontaneous mutations. We previously reported that sequential acquisition of mutations in the two-component regulatory systems vraSR and graRS was responsible for the VISA phenotype of strain Mu50. Here we report on the identification of a novel set of regulator mutations, a deletion mutation in two-component regulatory system walRK (synonyms, vicRK and yycFG), and a truncating mutation in a proteolytic regulatory gene, clpP, responsible for the raised vancomycin resistance in a laboratory-derived VISA strain, LR5P1-V3. The contributory effect of the two mutations to vancomycin resistance was confirmed by introducing the walK and clpP mutations into the vancomycin-susceptible parent strain N315LR5P1 by a gene replacement procedure. The vancomycin MIC of N315LR5P1 was raised from 1 to 2 mg/liter by the introduction of the walK or clpP mutation, but it was raised to 4 mg/liter by the introduction of both the walK and clpP mutations. The vancomycin MIC value of the double mutant was equivalent to that of strain LR5P1-V3. Like VISA clinical strains, LR5P1-V3 and the double mutant strain LR5P1walK*clpP* exhibited a thickened cell wall, slow growth, and decreased autolytic activity. Transcriptional profiles of the mutants with gene replacements demonstrated that introduction of both the walK and clpP mutations could alter expression of dozens or hundreds of genes, including those involved in cell envelope and cellular processes, intermediary metabolism, and information pathway. A mutation prevalence study performed on 39 worldwide clinical VISA strains showed that 61.5, 7.7, 10.3, and 20.5% of VISA strains harbored mutations in walRK, clpP, graRS, and vraSR, respectively. The mutation of walRK was most frequently carried by VISA strains. Together, these results suggested that the mutations of walK and clpP identified in LR5P1-V3 constitute a new combination of genetic events causing vancomycin resistance in Staphylococcus aureus. PMID- 21628540 TI - Genetic context and structural diversity of class 1 integrons from human commensal bacteria in a hospital intensive care unit. AB - Most surveys for class 1 integrons are at least partly predicated on PCR screening that targets integron conserved regions. However, class 1 integrons are structurally diverse, so dependence on conserved regions may lead to missing clinically relevant examples of class 1 integrons. Here, we surveyed a commensal population of bacteria from patients in an intensive care unit to identify class 1 integrons irrespective of their structure or genetic context. We identified several examples of class 1 integrons linked to complete Tn402-like or Tn402 hybrid transposition modules and diverse insertion points with respect to the inverted repeat IRi boundary. The diversity and abundance of class 1 integrons identified are such that many novel elements seen here would not have been identified by commonly used methods, and they revealed an additional level of complexity. PMID- 21628541 TI - In vitro efficacy of antibiotics commonly used to treat human plague against intracellular Yersinia pestis. AB - Yersinia pestis initiates infection as a facultative intracellular parasite in host macrophages; however, little is known about the efficacy of antibiotics commonly used to treat human plague against intracellular Y. pestis. Intracellular minimal bactericidal concentrations (MBCs) were determined using a high-throughput broth microdilution assay in which human THP-1 macrophage-like cells were infected with Y. pestis strain KIM6-2053.1+ and exposed to 2-fold serial dilutions of antibiotics for 24 h in 96-well plates. The numbers of CFU, upon which minimal bactericidal concentrations were based, were determined by counting "microcolonies" in wells of 96-well plates following lysis of tissue culture cells to release surviving Y. pestis, replica dilution, and plating in soft tryptic soy broth agar. For THP-1 cells, streptomycin and ciprofloxacin had comparable efficacies for intra- and extracellular Y. pestis, but the MBCs for chloramphenicol, gentamicin, doxycycline, and amoxicillin were two-, three-, four , and five 2-fold serial dilutions greater, respectively, for intracellular than for extracellular Y. pestis. During the initial stage of plague, intracellular Y. pestis may be less susceptible to antibiotic killing by particular antibiotics recommended for treatment of plague, such as gentamicin or doxycycline, whereas others, such as streptomycin and ciprofloxacin, may have similar efficacies against extracellular or intracellular Y. pestis. This may be of particular importance in the selection of antibiotics for prophylactic treatment in the case of a bioterrorism event. PMID- 21628542 TI - Antiviral efficacy upon administration of a HepDirect prodrug of 2'-C methylcytidine to hepatitis C virus-infected chimpanzees. AB - Hepatitis C virus (HCV) infects an estimated 170 million individuals worldwide, and the current standard of care, a combination of pegylated interferon alpha and ribavirin, is efficacious in achieving sustained viral response in ~50% of treated patients. Novel therapies under investigation include the use of nucleoside analog inhibitors of the viral RNA-dependent RNA polymerase. NM283, a 3'-valyl ester prodrug of 2'-C-methylcytidine, has demonstrated antiviral efficacy in HCV-infected patients (N. Afdhal et al., J. Hepatol. 46[Suppl. 1]:S5, 2007; N. Afdhal et al., J. Hepatol. 44[Suppl. 2]:S19, 2006). One approach to increase the antiviral efficacy of 2'-C-methylcytidine is to increase the concentration of the active inhibitory species, the 5'-triphosphate, in infected hepatocytes. HepDirect prodrug technology can increase intracellular concentrations of a nucleoside triphosphate in hepatocytes by introducing the nucleoside monophosphate into the cell, bypassing the initial kinase step that is often rate limiting. Screening for 2'-C-methylcytidine triphosphate levels in rat liver after oral dosing identified 1-[3,5-difluorophenyl]-1,3-propandiol as an efficient prodrug modification. To determine antiviral efficacy in vivo, the prodrug was administered separately via oral and intravenous dosing to two HCV infected chimpanzees. Circulating viral loads declined by ~1.4 log(10) IU/ml and by >3.6 log(10) IU/ml after oral and intravenous dosing, respectively. The viral loads rebounded after the end of dosing to predose levels. The results indicate that a robust antiviral response can be achieved upon administration of the prodrug. PMID- 21628543 TI - Tripropeptin C blocks the lipid cycle of cell wall biosynthesis by complex formation with undecaprenyl pyrophosphate. AB - Tripropeptin C (TPPC) is a naturally occurring cyclic lipodepsipeptide antibiotic produced by a Lysobacter sp. TPPC exhibits potent antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE), and penicillin-resistant Streptococcus pneumoniae. This antibiotic also inhibits the incorporation of N-acetylglucosamine into the peptidoglycan of S. aureus at a 50% inhibitory concentration (IC(50)) of 0.7 MUM, which is proportional to its MIC (0.87 MUM; equivalent to 1.0 MUg/ml). Treatment of exponential-phase S. aureus cells with TPPC resulted in accumulation of UDP MurNAc-pentapeptide in the cytoplasm. The antimicrobial activity of TPPC was weakened by the addition of prenyl pyrophosphates but not by prenyl phosphates, UDP-linked sugars, or the pentapeptide of peptidoglycan. The direct interaction between TPPC and undecaprenyl pyrophosphate (C(55)-PP) was observed by mass spectrometry and thin-layer chromatography analysis, indicating that TPPC can potentially inhibit C(55)-PP phosphatase activity, which plays a crucial role in the lipid cycle of peptidoglycan synthesis. As expected, TPPC inhibits this enzymatic reaction at an IC(50) of 0.03 to 0.1 MUM in vitro, as does bacitracin. From the analysis of accumulation of lipid carrier-related compounds, TPPC was found to cause the accumulation of C(55)-PP in situ, leading to the accumulation of a glycine-containing lipid intermediate. This suggested that the TPPC/C(55)-PP complex also inhibits the transglycosylation step or flippase activity, adding to the inhibition of C(55)-PP dephosphorylation. This mode of action is different from that of currently available drugs such as vancomycin, daptomycin, and bacitracin. PMID- 21628544 TI - Novel method for simultaneous quantification of phenotypic resistance to maturation, protease, reverse transcriptase, and integrase HIV inhibitors based on 3'Gag(p2/p7/p1/p6)/PR/RT/INT-recombinant viruses: a useful tool in the multitarget era of antiretroviral therapy. AB - Twenty-six antiretroviral drugs (ARVs), targeting five different steps in the life cycle of the human immunodeficiency virus type 1 (HIV-1), have been approved for the treatment of HIV-1 infection. Accordingly, HIV-1 phenotypic assays based on common cloning technology currently employ three, or possibly four, different recombinant viruses. Here, we describe a system to assess HIV-1 resistance to all drugs targeting the three viral enzymes as well as viral assembly using a single patient-derived, chimeric virus. Patient-derived p2-INT (gag-p2/NCp7/p1/p6/pol PR/RT/IN) products were PCR amplified as a single fragment (3,428 bp) or two overlapping fragments (1,657 bp and 2,002 bp) and then recombined into a vector containing a near-full-length HIV-1 genome with the Saccharomyces cerevisiae uracil biosynthesis gene (URA3) replacing the 3,428 bp p2-INT segment (Dudley et al., Biotechniques 46:458-467, 2009). P2-INT-recombinant viruses were employed in drug susceptibility assays to test the activity of protease (PI), nucleoside/nucleotide reverse transcriptase (NRTI), nonnucleoside reverse transcriptase (NNRTI), and integrase strand-transfer (INSTI) inhibitors. Using a single standardized test (ViralARTS HIV), this new technology permits the rapid and automated quantification of phenotypic resistance for all known and candidate antiretroviral drugs targeting all viral enzymes (PR, RT, including polymerase and RNase H activities, and IN), some of the current and potential assembly inhibitors, and any drug targeting Pol or Gag precursor cleavage sites (relevant for PI and maturation inhibitors) This novel assay may be instrumental (i) in the development and clinical assessment of novel ARV drugs and (ii) to monitor patients failing prior complex treatment regimens. PMID- 21628546 TI - Modulation of clock gene expression by the transcriptional coregulator receptor interacting protein 140 (RIP140). AB - Circadian rhythms are generated in central and peripheral tissues by an intracellular oscillating timing mechanism known as the circadian clock. Several lines of evidence show a strong and bidirectional interplay between metabolism and circadian rhythms. Receptor interacting protein 140 (RIP140) is a coregulator for nuclear receptors and other transcription factors that represses catabolic pathways in metabolic tissues. Although RIP140 functions as a corepressor for most nuclear receptors, mounting evidence points to RIP140 as a dual coregulator that can repress or activate different sets of genes. Here, we demonstrate that RIP140 mRNA and protein levels are under circadian regulation and identify RIP140 as a modulator of clock gene expression, suggesting that RIP140 can participate in a feedback mechanism affecting the circadian clock. We show that the absence of RIP140 disturbs the basal levels of BMAL1 and other clock genes, reducing the amplitude of their oscillations. In addition, we demonstrate that RIP140 is recruited to retinoid-related orphan receptor (ROR) binding sites on the BMAL1 promoter, directly interacts with RORalpha, and increases transcription from the BMAL1 promoter in a RORalpha-dependent manner. These results indicate that RIP140 is not only involved in metabolic control but also acts as a coactivator for RORalpha, influencing clock gene expression. PMID- 21628547 TI - Effects of vasoactive intestinal peptide genotype on circadian gene expression in the suprachiasmatic nucleus and peripheral organs. AB - The neuropeptide vasoactive intestinal polypeptide (VIP) has emerged as a key candidate molecule mediating the synchronization of rhythms in clock gene expression within the suprachiasmatic nucleus (SCN). In addition, neurons expressing VIP are anatomically well positioned to mediate communication between the SCN and peripheral oscillators. In this study, we examined the temporal expression profile of 3 key circadian genes: Per1, Per2 , and Bmal1 in the SCN, the adrenal glands and the liver of mice deficient for the Vip gene (VIP KO), and their wild-type counterparts. We performed these measurements in mice held in a light/dark cycle as well as in constant darkness and found that rhythms in gene expression were greatly attenuated in the VIP-deficient SCN. In the periphery, the impact of the loss of VIP varied with the tissue and gene measured. In the adrenals, rhythms in Per1 were lost in VIP-deficient mice, while in the liver, the most dramatic impact was on the phase of the diurnal expression rhythms. Finally, we examined the effects of the loss of VIP on ex vivo explants of the same central and peripheral oscillators using the PER2::LUC reporter system. The VIP-deficient mice exhibited low amplitude rhythms in the SCN as well as altered phase relationships between the SCN and the peripheral oscillators. Together, these data suggest that VIP is critical for robust rhythms in clock gene expression in the SCN and some peripheral organs and that the absence of this peptide alters both the amplitude of circadian rhythms as well as the phase relationships between the rhythms in the SCN and periphery. PMID- 21628545 TI - Susceptibility of Streptococcus pneumoniae to fluoroquinolones in Canada. AB - Ciprofloxacin, the first fluoroquinolone to be used to treat lower respiratory tract infections (LRTI), demonstrates poor potency against Streptococcus pneumoniae, and its use has been associated with the emergence of resistance. During the last decade, fluoroquinolones with enhanced in vitro activity against S. pneumoniae have replaced ciprofloxacin for the treatment of LRTI. Here, we analyzed the impact of more active fluoroquinolone usage on pneumococci by examining the fluoroquinolone usage, prevalence of fluoroquinolone resistance, and mutations in the genes that encode the major target sites for the fluoroquinolones (gyrA and parC) in pneumococcal isolates collected in Canada wide surveillance. A total of 26,081 isolates were collected between 1998 and 2009. During this time period, total per capita outpatient use of fluoroquinolones increased from 64 to 96 prescriptions per 1,000 persons per year. The proportion of prescriptions for respiratory tract infection that were for fluoroquinolones increased from 5.9% to 10.7%, but the distribution changed: the proportion of prescriptions for ciprofloxacin decreased from 5.3% to 0.5%, and those for levofloxacin or moxifloxacin increased from 1.5% in 1999 to 5.9% in 2009. The prevalence of ciprofloxacin resistance (MIC >= 4 MUg/ml), levofloxacin resistance, and moxifloxacin resistance remained unchanged at <2%. Multivariable analyses showed that prevalence of mutations known to be associated with reduced susceptibility to fluoroquinolones did not change during the surveillance period. If fluoroquinolone therapy is required, the preferential use of fluoroquinolones with enhanced pneumococcal activity to treat pneumococcal infections may slow the emergence of resistance in S. pneumoniae. PMID- 21628548 TI - Cyclic AMP signaling control of action potential firing rate and molecular circadian pacemaking in the suprachiasmatic nucleus. AB - Circadian pacemaking in suprachiasmatic nucleus (SCN) neurons revolves around transcriptional/posttranslational feedback loops, driven by protein products of "clock" genes. These loops are synchronized and sustained by intercellular signaling, involving vasoactive intestinal peptide (VIP) via its VPAC2 receptor, which positively regulates cAMP synthesis. In turn, SCN cells communicate circadian time to the brain via a daily rhythm in electrophysiological activity. To investigate the mechanisms whereby VIP/VPAC2/cAMP signaling controls SCN molecular and electrical pacemaking, we combined bioluminescent imaging of circadian gene expression and whole-cell electrophysiology in organotypic SCN slices. As a potential direct target of cAMP, we focused on hyperpolarization activated, cyclic nucleotide-gated (HCN) channels. Mutations of VIP-ergic signaling compromised the SCN molecular pacemaker, diminishing the amplitude and intercellular synchrony of circadian gene expression. These deficits were transiently reversed by elevation of cAMP. Similarly, cellular synchrony in electrical firing rates was lost in SCN slices lacking the VPAC2 receptor for VIP. Whole-cell current-clamp recordings in wild-type (WT) slices revealed voltage responses shaped by the conductance I(h), which is mediated by HCN channel activity. The influence of I(h) on voltage responses showed a modest peak in early circadian day, identifying HCN channels as a putative mediator of cAMP dependent circadian effects on firing rate. I(h), however, was unaffected by loss of VIP-ergic signaling in VPAC2-null slices, and inhibition of cAMP synthesis had no discernible effect on I(h) but did suppress gene expression and SCN firing rates. Moreover, only sustained but not acute, pharmacological blockade of HCN channels reduced action potential (AP) firing. Thus, our evidence suggests that in the SCN, cAMP-mediated signaling is not a principal regulator of HCN channel function and that HCN is not a determinant of AP firing rate. VIP/cAMP-dependent signaling sustains the SCN molecular oscillator and action potential firing via mechanisms yet to be identified. PMID- 21628549 TI - Resilience of circadian pacemaker development in hamsters. AB - Disruptions of circadian rhythms have been linked to a wide range of pathologies from sleep disorders to cancer. The extent to which disruptions of circadian rhythms during development contribute to later conditions is not known. The present study tested the hypothesis that functional properties of the central circadian pacemaker, the suprachiasmatic nucleus (SCN), are affected by abnormal entrainment during development. The SCN is specialized for the generation of robust rhythms, for direct and indirect output to physiological and behavioral systems, and for entrainment to light/dark cycles via direct retinal input. It consists of thousands of neurons and glia with distinct phenotypes and has subdivisions delineated by both anatomical and functional criteria. In rodents, SCN rhythms develop within days after SCN cells are produced and before many other aspects of differentiation, such as synaptogenesis, are complete. We demonstrated that around the time of birth, the hamster SCN in vivo can undergo repeated phase shifts by a dopamine D(1) receptor agonist (SKF-38393). For 2 days before and 2 days after birth, one group of hamsters received regular exposure to the drug at the same time of day, while another group was exposed at varying times to induce repeated phase shifts. Free-running and entrained activity rhythms were compared between the groups at different ages after weaning. Repeated phase shifts during SCN development had a significant effect on free running period measured immediately after weaning. This effect was eliminated by subsequent entrainment to a light/dark cycle, indicating that the effect was not permanent. These and other results suggest that SCN development required for functional properties such as free-running period is resilient to perturbation. PMID- 21628550 TI - Refeeding after fasting elicits insulin-dependent regulation of Per2 and Rev erbalpha with shifts in the liver clock. AB - The mammalian circadian clock is known to be entrained by both a daily light-dark cycle and daily feeding cycle. However, the mechanisms of feeding-induced entrainment are not as fully understood as those of light entrainment. To elucidate the first step of entrainment of the liver clock, we identified the circadian clock gene(s) that show both phase advance and acute change of gene expression during the early term of the daytime refeeding schedule in mice. The expressions of liver Per2 and Rev-erbalpha genes were phase-advanced within 1 day of refeeding. Additionally, the upregulation of Per2 mRNA and down-regulation of Rev-erbalpha mRNA were induced within 2 hours, not only by food intake but also by insulin injection in intact mice. These expression changes by food intake were not revealed in streptozotocin-treated insulin-deficient mice, but insulin injection was able to recover the impairment of Per2 and Rev-erbalpha gene expression. Furthermore, we demonstrated using an ex vivo luciferase monitoring system that insulin injection during the daytime causes a phase advance of liver Per2 expression rhythm in Per2::luciferase knock-in mice. In embryonic fibroblasts from Per2::luciferase knock-in mice, insulin infusion caused an acute increase of Per2 gene expression and a similar phase advance of Per2 expression rhythm. Our results indicate that an acute change of Per2 and Rev-erbalpha gene expression mediated by refeeding-induced insulin secretion is a critical step mediating the early phase of feeding-induced entrainment of the liver clock. PMID- 21628551 TI - Torpor in dark times: patterns of heterothermy are associated with the lunar cycle in a nocturnal bird. AB - Many studies have shown that endotherms become more heterothermic when the costs of thermoregulation are high and/or when limited energy availability constrains thermoregulatory capacity. However, the roles of many ecological variables, including constraints on foraging opportunities and/or success, remain largely unknown. To test the prediction that thermoregulatory patterns should be related to foraging opportunities in a heterothermic endotherm, we examined the relationship between the lunar cycle and heterothermy in Freckled Nightjars (Caprimulgus tristigma), which are visually orienting, nocturnal insectivores that are dependent on ambient light to forage. This model system provides an opportunity to assess whether variation in foraging opportunities influences the expression of heterothermy. The nightjars were active and foraged for insects when moonlight was available but became inactive and heterothermic in the absence of moonlight. Lunar illumination was a much stronger predictor of the magnitude of heterothermic responses than was air temperature (T(a)). Our data suggest that heterothermy was strongly related to variation in foraging opportunities associated with the lunar cycle, even though food abundance appeared to remain relatively high throughout the study period. Patterns of thermoregulation in this population of Freckled Nightjars provide novel insights into the environmental and ecological determinants of heterothermy, with the lunar cycle, and not T(a), being the strongest predictor of torpor use. PMID- 21628552 TI - Effects of light on cognitive brain responses depend on circadian phase and sleep homeostasis. AB - Light is a powerful modulator of cognition through its long-term effects on circadian rhythmicity and direct effects on brain function as identified by neuroimaging. How the direct impact of light on brain function varies with wavelength of light, circadian phase, and sleep homeostasis, and how this differs between individuals, is a largely unexplored area. Using functional MRI, we compared the effects of 1 minute of low-intensity blue (473 nm) and green light (527 nm) exposures on brain responses to an auditory working memory task while varying circadian phase and status of the sleep homeostat. Data were collected in 27 subjects genotyped for the PER3 VNTR (12 PER3(5/5) and 15 PER3(4/4) ) in whom it was previously shown that the brain responses to this task, when conducted in darkness, depend on circadian phase, sleep homeostasis, and genotype. In the morning after sleep, blue light, relative to green light, increased brain responses primarily in the ventrolateral and dorsolateral prefrontal cortex and in the intraparietal sulcus, but only in PER3(4/4) individuals. By contrast, in the morning after sleep loss, blue light increased brain responses in a left thalamofrontoparietal circuit to a larger extent than green light, and only so in PER3(5/5) individuals. In the evening wake maintenance zone following a normal waking day, no differential effect of 1 minute of blue versus green light was observed in either genotype. Comparison of the current results with the findings observed in darkness indicates that light acts as an activating agent particularly under those circumstances in which and in those individuals in whom brain function is jeopardized by an adverse circadian phase and high homeostatic sleep pressure. PMID- 21628553 TI - Predominance of distal skin temperature changes at sleep onset across menstrual and circadian phases. AB - Menstrual cycle-associated changes in reproductive hormones affect body temperature in women. We aimed to characterize the interaction between the menstrual, circadian, and scheduled sleep-wake cycles on body temperature regulation. Eight females entered the laboratory during the midfollicular (MF) and midluteal (ML) phases of their menstrual cycle for an ultradian sleep-wake cycle procedure, consisting of 36 cycles of 60-minute wake episodes alternating with 60-minute nap opportunities, in constant bed-rest conditions. Core body temperature (CBT) and distal skin temperature (DT) were recorded and used to calculate a distal-core gradient (DCG). Melatonin, sleep, and subjective sleepiness were also recorded. The circadian variation of DT and DCG was not affected by menstrual phase. DT and DCG showed rapid, large nap episode-dependent increases, whereas CBT showed slower, smaller nap episode-dependent decreases. DCG values were significantly reduced for most of the wake episode in an overall 60-minute wake/60-minute nap cycle during ML compared to MF, but these differences were eliminated at the wake-to-nap lights-out transition. Nap episode dependent decreases in CBT were further modulated as a function of both circadian and menstrual factors, with nap episode-dependent deceases occurring more prominently during the late afternoon/evening in ML, whereas nap episode dependent DT and DCG increases were not significantly affected by menstrual phase but only circadian phase. Circadian rhythms of melatonin secretion, DT, and DCG were significantly phase-advanced relative to CBT and sleep propensity rhythms. This study explored how the thermoregulatory system is influenced by an interaction between circadian phase and vigilance state and how this is further modulated by the menstrual cycle. Current results agree with the thermophysiological cascade model of sleep and indicate that despite increased CBT during ML, heat loss mechanisms are maintained at a similar level during nap episodes, which may allow for comparable circadian sleep propensity rhythms between menstrual phases. PMID- 21628554 TI - Resveratrol dietary supplementation shortens the free-running circadian period and decreases body temperature in a prosimian primate. AB - Resveratrol (RSV) is a dietary polyphenolic compound with several positive effects on metabolic functions and longevity. We tested the effect of RSV on the circadian clock in a nonhuman primate, the gray mouse lemur. The impact of a 2 week dietary supplementation of RSV on the rhythms of locomotor activity and body temperature in constant dark conditions (DD) was investigated in young (n = 7) and old (n = 6) animals. RSV supplementation followed 2 weeks in DD under normal diet (CTL). In both young and old animals receiving RSV, we observed a shortening of the free-running period compared to those under CTL (-15 minutes in young animals and -45 minutes in old animals), accompanied by a lower mean body temperature in both age groups and decreased locomotor activity in young animals. Thus, RSV is a food component capable of influencing a primate's circadian clock. This property may be of interest clinically in the context of the treatment of circadian disruption and in the context of the effects of RSV ingestion on health. PMID- 21628555 TI - Human diurnal preference and circadian rhythmicity are not associated with the CLOCK 3111C/T gene polymorphism. AB - Genetic association studies of the CLOCK 3111C/T polymorphism and diurnal preference have yielded conflicting results since the first report that the 3111C allele was associated with eveningness. The goal of the present study was to investigate the association of this polymorphism with diurnal preference and circadian physiology in a group of 179 individuals, by comparing the frequency of the 3111C allele to diurnal preference, habitual sleep timing, circadian phase markers, and circadian period. We did not find a significant association between this allele and morningness/eveningness or any circadian marker. PMID- 21628556 TI - Postsynaptic P/Q-type Ca2+ channel in Purkinje cell mediates synaptic competition and elimination in developing cerebellum. AB - Neural circuits are initially redundant but rearranged through activity-dependent synapse elimination during postnatal development. This process is crucial for shaping mature neural circuits and for proper brain function. At birth, Purkinje cells (PCs) in the cerebellum are innervated by multiple climbing fibers (CFs) with similar synaptic strengths. During postnatal development, a single CF is selectively strengthened in each PC through synaptic competition, the strengthened single CF undergoes translocation to a PC dendrite, and massive elimination of redundant CF synapses follows. To investigate the cellular mechanisms of this activity-dependent synaptic refinement, we generated mice with PC-selective deletion of the Ca(v)2.1 P/Q-type Ca(2+) channel, the major voltage dependent Ca(2+) channel in PCs. In the PC-selective Ca(v)2.1 knockout mice, Ca(2+) transients induced by spontaneous CF inputs are markedly reduced in PCs in vivo. Not a single but multiple CFs were equally strengthened in each PC from postnatal day 5 (P5) to P8, multiple CFs underwent translocation to PC dendrites, and subsequent synapse elimination until around P12 was severely impaired. Thus, P/Q-type Ca(2+) channels in postsynaptic PCs mediate synaptic competition among multiple CFs and trigger synapse elimination in developing cerebellum. PMID- 21628557 TI - Canopy1, a positive feedback regulator of FGF signaling, controls progenitor cell clustering during Kupffer's vesicle organogenesis. AB - The assembly of progenitor cells is a crucial step for organ formation during vertebrate development. Kupffer's vesicle (KV), a key organ required for the left right asymmetric body plan in zebrafish, is generated from a cluster of ~20 dorsal forerunner cells (DFCs). Although several genes are known to be involved in KV formation, how DFC clustering is regulated and how cluster formation then contributes to KV formation remain unclear. Here we show that positive feedback regulation of FGF signaling by Canopy1 (Cnpy1) controls DFC clustering. Cnpy1 positively regulates FGF signals within DFCs, which in turn promote Cadherin1 mediated cell adhesion between adjacent DFCs to sustain cell cluster formation. When this FGF positive feedback loop is disrupted, the DFC cluster fails to form, eventually leading to KV malformation and defects in the establishment of laterality. Our results therefore uncover both a previously unidentified role of FGF signaling during vertebrate organogenesis and a regulatory mechanism underlying cell cluster formation, which is an indispensable step for formation of a functional KV and establishment of the left-right asymmetric body plan. PMID- 21628558 TI - Complementary assays reveal a relationship between HIV-1 uncoating and reverse transcription. AB - During the early stages of HIV-1 replication the conical capsid composed of p24(CA) protein dissociates from the rest of the cytoplasmic viral complex by a process called uncoating. Although proper uncoating is known to be required for HIV-1 infection, many questions remain about the timing and factors involved in the process. Here we have used two complementary assays to study the process of uncoating in HIV-1-infected cells, specifically looking at the timing of uncoating and its relationship to reverse transcription. We developed a fluorescent microscopy-based uncoating assay that detects the association of p24(CA) with HIV-1 viral complexes in cells. We also used an owl monkey kidney (OMK) cell assay that is based on timed TRIM-CypA-mediated restriction of HIV-1 replication. Results from both assays indicate that uncoating is initiated within 1 h of viral fusion. In addition, treatment with the reverse transcriptase inhibitor nevirapine delayed uncoating in both assays. Analysis of reverse transcription products in OMK cells revealed that the generation of early reverse transcription products coincides with the timing of uncoating in these assays. Collectively, these results suggest that some aspect of reverse transcription has the ability to influence the kinetics of uncoating. PMID- 21628559 TI - Defects in coatomer protein I (COPI) transport cause blood feeding-induced mortality in Yellow Fever mosquitoes. AB - Blood feeding by vector mosquitoes provides the entry point for disease pathogens and presents an acute metabolic challenge that must be overcome to complete the gonotrophic cycle. Based on recent data showing that coatomer protein I (COPI) vesicle transport is involved in cellular processes beyond Golgi-endoplasmic reticulum retrograde protein trafficking, we disrupted COPI functions in the Yellow Fever mosquito Aedes aegypti to interfere with blood meal digestion. Surprisingly, we found that decreased expression of the gammaCOPI coatomer protein led to 89% mortality in blood-fed mosquitoes by 72 h postfeeding compared with 0% mortality in control dsRNA-injected blood-fed mosquitoes and 3% mortality in gammaCOPI dsRNA-injected sugar-fed mosquitoes. Similar results were obtained using dsRNA directed against five other COPI coatomer subunits (alpha, beta, beta', delta, and zeta). We also examined midgut tissues by EM, quantitated heme in fecal samples, and characterized feeding-induced protein expression in midgut, fat body, and ovary tissues of COPI-deficient mosquitoes. We found that COPI defects disrupt epithelial cell membrane integrity, stimulate premature blood meal excretion, and block induced expression of several midgut protease genes. To study the role of COPI transport in ovarian development, we injected gammaCOPI dsRNA after blood feeding and found that, although blood digestion was normal, follicles in these mosquitoes were significantly smaller by 48 h postinjection and lacked eggshell proteins. Together, these data show that COPI functions are critical to mosquito blood digestion and egg maturation, a finding that could also apply to other blood-feeding arthropod vectors. PMID- 21628560 TI - Tropical amphibian populations experience higher disease risk in natural habitats. AB - Habitat loss and disease are main drivers of global amphibian declines, yet the interaction between them remains largely unexplored. Here we show that paradoxically, habitat loss is negatively associated with occurrence, prevalence, and infection intensity of the chytrid fungus Batrachochytrium dendrobatidis (Bd) in amphibian populations in the tropics. At a large spatial scale, increased habitat loss predicted lower disease risk in amphibian populations across Costa Rica and eastern Australia, even after jointly considering the effect of potential biotic and abiotic correlates. Lower host-species richness and suboptimal microclimates for Bd in disturbed habitats are potential mechanisms underlying this pattern. Furthermore, we found that anthropogenic deforestation practices biased to lowlands and natural vegetation remaining in inaccessible highlands explain increased Bd occurrence at higher elevations. At a smaller spatial scale, holding constant elevation, latitude, and macroclimate, we also found a negative relationship between habitat loss, and both Bd prevalence and infection intensity in frog populations in two landscapes of the Brazilian Atlantic Forest. Our results indicate that amphibians will be disproportionately affected by emerging diseases in pristine environments, and that, paradoxically, disturbed habitats may act as shelters from disease, but only for the very few species that can tolerate deforestation. Thus, tropical amphibian faunas are threatened both by destruction of natural habitats as well as increased disease in pristine forests. To curb further extinctions and develop effective mitigation and restoration programs we must look to interactions between habitat loss and disease, the two main factors at the root of global amphibian declines. PMID- 21628561 TI - Lipooligosaccharide is required for the generation of infectious elementary bodies in Chlamydia trachomatis. AB - Lipopolysaccharides (LPS) and lipooligosaccharides (LOS) are the main lipid components of bacterial outer membranes and are essential for cell viability in most Gram-negative bacteria. Here we show that small molecule inhibitors of LpxC [UDP-3-O-(R-3-hydroxymyristoyl)-GlcNAc deacetylase], the enzyme that catalyzes the first committed step in the biosynthesis of lipid A, block the synthesis of LOS in the obligate intracellular bacterial pathogen Chlamydia trachomatis. In the absence of LOS, Chlamydia remains viable and establishes a pathogenic vacuole ("inclusion") that supports robust bacterial replication. However, bacteria grown under these conditions were no longer infectious. In the presence of LpxC inhibitors, replicative reticulate bodies accumulated in enlarged inclusions but failed to express selected late-stage proteins and transition to elementary bodies, a Chlamydia developmental form that is required for invasion of mammalian cells. These findings suggest the presence of an outer membrane quality control system that regulates Chlamydia developmental transition to infectious elementary bodies and highlights the potential application of LpxC inhibitors as unique class of antichlamydial agents. PMID- 21628562 TI - Ancient DNA reveals male diffusion through the Neolithic Mediterranean route. AB - The Neolithic is a key period in the history of the European settlement. Although archaeological and present-day genetic data suggest several hypotheses regarding the human migration patterns at this period, validation of these hypotheses with the use of ancient genetic data has been limited. In this context, we studied DNA extracted from 53 individuals buried in a necropolis used by a French local community 5,000 y ago. The relatively good DNA preservation of the samples allowed us to obtain autosomal, Y-chromosomal, and/or mtDNA data for 29 of the 53 samples studied. From these datasets, we established close parental relationships within the necropolis and determined maternal and paternal lineages as well as the absence of an allele associated with lactase persistence, probably carried by Neolithic cultures of central Europe. Our study provides an integrative view of the genetic past in southern France at the end of the Neolithic period. Furthermore, the Y-haplotype lineages characterized and the study of their current repartition in European populations confirm a greater influence of the Mediterranean than the Central European route in the peopling of southern Europe during the Neolithic transition. PMID- 21628563 TI - c-Met signaling induces a reprogramming network and supports the glioblastoma stem-like phenotype. AB - The tyrosine kinase c-Met promotes the formation and malignant progression of multiple cancers. It is well known that c-Met hyperactivation increases tumorigenicity and tumor cell resistance to DNA damaging agents, properties associated with tumor-initiating stem cells. However, a link between c-Met signaling and the formation and/or maintenance of neoplastic stem cells has not been previously identified. Here, we show that c-Met is activated and functional in glioblastoma (GBM) neurospheres enriched for glioblastoma tumor-initiating stem cells and that c-Met expression/function correlates with stem cell marker expression and the neoplastic stem cell phenotype in glioblastoma neurospheres and clinical glioblastoma specimens. c-Met activation was found to induce the expression of reprogramming transcription factors (RFs) known to support embryonic stem cells and induce differentiated cells to form pluripotent stem (iPS) cells, and c-Met activation counteracted the effects of forced differentiation in glioblastoma neurospheres. Expression of the reprogramming transcription factor Nanog by glioblastoma cells is shown to mediate the ability of c-Met to induce the stem cell characteristics of neurosphere formation and neurosphere cell self-renewal. These findings show that c-Met enhances the population of glioblastoma stem cells (GBM SCs) via a mechanism requiring Nanog and potentially other c-Met-responsive reprogramming transcription factors. PMID- 21628564 TI - Exploring the contribution and significance of animal protein in the diet of bonobos by stable isotope ratio analysis of hair. AB - In primates, age, sex, and social status can strongly influence access to food resources. In Pan, these criteria are assumed to influence access to vertebrate meat. However, the significance of meat in terms of its role in the nutrition of Pan is still debated. Here we present a study using stable carbon and nitrogen isotope ratios in hair samples from habituated, wild bonobos (Pan paniscus) to explore these issues. Over a period of 5 mo hair samples were collected from fresh bonobo nests at LuiKotale, Democratic Republic of Congo. Hair samples were assigned to known individuals and were of sufficient length to allow the evaluation of isotopic variation over several months. Samples of plant foods and sympatric fauna were also analyzed. The delta(13)C and delta(15)N results of the bonobo hair were remarkably homogeneous over time and for the group as a whole. There are no differences in diet between the sexes. Within the group of males, however, there was a positive correlation between dominance status and delta(15)N. The isotopic data indicate that the contribution of fauna to bonobo diet is marginal and that plant food is the dietary protein source. In only some cases did elevated delta(15)N hair values correlate with observed faunivory and not correspond to the delta(15)N measured in the dominant plant foods. Given the large variation in hunting and meat eating of Pan across the African continent, the detection of seasonal changes in faunivory by elevated delta(15)N values in sectioned ape hair is a promising approach. PMID- 21628565 TI - Enhancement of CA3 hippocampal network activity by activation of group II metabotropic glutamate receptors. AB - Impaired function or expression of group II metabotropic glutamate receptors (mGluRIIs) is observed in brain disorders such as schizophrenia. This class of receptor is thought to modulate activity of neuronal circuits primarily by inhibiting neurotransmitter release. Here, we characterize a postsynaptic excitatory response mediated by somato-dendritic mGluRIIs in hippocampal CA3 pyramidal cells and in stratum oriens interneurons. The specific mGluRII agonists DCG-IV or LCCG-1 induced an inward current blocked by the mGluRII antagonist LY341495. Experiments with transgenic mice revealed a significant reduction of the inward current in mGluR3(-/-) but not in mGluR2(-/-) mice. The excitatory response was associated with periods of synchronized activity at theta frequency. Furthermore, cholinergically induced network oscillations exhibited decreased frequency when mGluRIIs were blocked. Thus, our data indicate that hippocampal responses are modulated not only by presynaptic mGluRIIs that reduce glutamate release but also by postsynaptic mGluRIIs that depolarize neurons and enhance CA3 network activity. PMID- 21628566 TI - Monoubiquitin-dependent endocytosis of the iron-regulated transporter 1 (IRT1) transporter controls iron uptake in plants. AB - Plants take up iron from the soil using the iron-regulated transporter 1 (IRT1) high-affinity iron transporter at the root surface. Sophisticated regulatory mechanisms allow plants to tightly control the levels of IRT1, ensuring optimal absorption of essential but toxic iron. Here, we demonstrate that overexpression of Arabidopsis thaliana IRT1 leads to constitutive IRT1 protein accumulation, metal overload, and oxidative stress. IRT1 is unexpectedly found in trans-Golgi network/early endosomes of root hair cells, and its levels and localization are unaffected by iron nutrition. Using pharmacological approaches, we show that IRT1 cycles to the plasma membrane to perform iron and metal uptake at the cell surface and is sent to the vacuole for proper turnover. We also prove that IRT1 is monoubiquitinated on several cytosol-exposed residues in vivo and that mutation of two putative monoubiquitination target residues in IRT1 triggers stabilization at the plasma membrane and leads to extreme lethality. Together, these data suggest a model in which monoubiquitin-dependent internalization/sorting and turnover keep the plasma membrane pool of IRT1 low to ensure proper iron uptake and to prevent metal toxicity. More generally, our work demonstrates the existence of monoubiquitin-dependent trafficking to lytic vacuoles in plants and points to proteasome-independent turnover of plasma membrane proteins. PMID- 21628567 TI - Retraction for "Properdin homeostasis requires turnover of the alternative complement pathway". PMID- 21628568 TI - Approach to discover T- and B-cell antigens of intracellular pathogens applied to the design of Chlamydia trachomatis vaccines. AB - Natural immunity against obligate and/or facultative intracellular pathogens is usually mediated by both humoral and cellular immunity. The identification of those antigens stimulating both arms of the immune system is instrumental for vaccine discovery. Although high-throughput technologies have been applied for the discovery of antibody-inducing antigens, few examples of their application for T-cell antigens have been reported. We describe how the compilation of the immunome, here defined as the pool of immunogenic antigens inducing T- and B-cell responses in vivo, can lead to vaccine candidates against Chlamydia trachomatis. We selected 120 C. trachomatis proteins and assessed their immunogenicity using two parallel high-throughput approaches. Protein arrays were generated and screened with sera from C. trachomatis-infected patients to identify antibody inducing antigens. Splenocytes from C. trachomatis-infected mice were stimulated with 79 proteins, and the frequency of antigen-specific CD4(+)/IFN-gamma(+) T cells was analyzed by flow cytometry. We identified 21 antibody-inducing antigens, 16 CD4(+)/IFN-gamma(+)-inducing antigens, and five antigens eliciting both types of responses. Assessment of their protective activity in a mouse model of Chlamydia muridarum lung infection led to the identification of seven antigens conferring partial protection when administered with LTK63/CpG adjuvant. Protection was largely the result of cellular immunity as assessed by CD4(+) T cell depletion. The seven antigens provided robust additive protection when combined in four-antigen combinations. This study paves the way for the development of an effective anti-Chlamydia vaccine and provides a general approach for the discovery of vaccines against other intracellular pathogens. PMID- 21628569 TI - Unraveling the distributed neural code of facial identity through spatiotemporal pattern analysis. AB - Face individuation is one of the most impressive achievements of our visual system, and yet uncovering the neural mechanisms subserving this feat appears to elude traditional approaches to functional brain data analysis. The present study investigates the neural code of facial identity perception with the aim of ascertaining its distributed nature and informational basis. To this end, we use a sequence of multivariate pattern analyses applied to functional magnetic resonance imaging (fMRI) data. First, we combine information-based brain mapping and dynamic discrimination analysis to locate spatiotemporal patterns that support face classification at the individual level. This analysis reveals a network of fusiform and anterior temporal areas that carry information about facial identity and provides evidence that the fusiform face area responds with distinct patterns of activation to different face identities. Second, we assess the information structure of the network using recursive feature elimination. We find that diagnostic information is distributed evenly among anterior regions of the mapped network and that a right anterior region of the fusiform gyrus plays a central role within the information network mediating face individuation. These findings serve to map out and characterize a cortical system responsible for individuation. More generally, in the context of functionally defined networks, they provide an account of distributed processing grounded in information-based architectures. PMID- 21628570 TI - Pathway-specific engagement of ephrinA5-EphA4/EphA5 system of the substantia nigra pars reticulata in cocaine-induced responses. AB - The nucleus accumbens (NAc) serves as a key neural substrate that controls acute and adaptive behavioral responses to cocaine administration. In this circuit, inputs from the NAc are transmitted through two parallel pathways, named the direct and indirect pathways, and converge at the substantia nigra pars reticulata (SNr). Our previous study using reversible neurotransmission blocking (RNB) of each pathway revealed that the dual stimulation of the SNr by both pathways is necessary for the acute response, but that the direct pathway predominantly controls the adaptive response to repeated cocaine administration. This study aimed at exploring the pathway-specific mechanism of cocaine actions at the convergent SNr. We examined a genome-wide expression profile of the SNr of three types of experimental mice: the direct pathway-blocked D-RNB mice, the indirect pathway-blocked I-RNB mice, and wild-type mice. We identified the up regulation of ephrinA5, EphA4, and EphA5 specific to D-RNB mice during both acute and adaptive responses to cocaine administration. The activation by EphA4 and EphA5 in the SNr of wild-type mice by use of the immunoadhesin technique suppressed the adaptive response to repeated cocaine administration. Furthermore, cocaine exposure stimulated the phosphorylation of Erk1/2 in ephrinA5-expressing SNr cells in a direct pathway-dependent manner. The results have demonstrated that the ephrinA5-EphA4/EphA5 system plays an important role in the direct pathway-dependent regulation of the SNr in both acute and adaptive cocaine responses and would provide valuable therapeutic targets of cocaine addiction. PMID- 21628571 TI - Mannose receptor polyubiquitination regulates endosomal recruitment of p97 and cytosolic antigen translocation for cross-presentation. AB - The molecular mechanisms regulating noncanonical protein transport across cellular membranes are poorly understood. Cross-presentation of exogenous antigens on MHC I molecules by dendritic cells (DCs) generally requires antigen translocation from the endosomal compartment into the cytosol for proteasomal degradation. In this study, we demonstrate that such translocation is controlled by the endocytic receptor and regulated by ubiquitination. Antigens internalized by the mannose receptor (MR), an endocytic receptor that targets its ligands specifically toward cross-presentation, were translocated into the cytosol only after attachment of a lysin48-linked polyubiquitin chain to the cytosolic region of the MR. Furthermore, we identify TSG101 as a central regulator of MR ubiquitination and antigen translocation. Importantly, we demonstrate that MR polyubiquitination mediates the recruitment of p97, a member of the ER-associated degradation machinery that provides the driving force for antigen translocation, toward the endosomal membrane, proving the central role of the endocytic receptor and its ubiquitination in antigen translocation. PMID- 21628572 TI - Circadian clock disruption improves the efficacy of chemotherapy through p73 mediated apoptosis. AB - The circadian clock in mammalian organisms is generated by a transcription translation feedback loop that controls many biochemical pathways at the cellular level and physiology and behavior at the organismal level. Cryptochrome (Cry) is a key protein in the negative arm of the transcription-translation feedback loop. It has been found that Cry mutation in cells with p53-null genotype increased their sensitivity to apoptosis by genotoxic agents. Here we show that this increased sensitivity is due to up-regulation of the p53 gene family member p73 in response to DNA damage. As a consequence, when tumors arising from oncogenic Ras-transformed p53(-/-) and p53(-/-)Cry1(-/-)Cry2(-/-) cells are treated with the anticancer drug oxaliplatin, p53(-/-) tumors continue to grow whereas p53(-/ )Cry1(-/-)Cry2(-/-) tumors exhibit extensive apoptosis and stop growing. This finding provides a mechanistic foundation for overcoming the resistance of p53 deficient tumor cells to apoptosis induced by DNA-damaging agents and suggests that disruption of cryptochrome function may increase the sensitivity of tumors with p53 mutation to chemotherapy. PMID- 21628573 TI - Mammary epithelial-specific disruption of c-Src impairs cell cycle progression and tumorigenesis. AB - The tyrosine kinase c-Src is activated in a large proportion of breast cancers, in which it is thought to play a key role in promoting the malignant phenotype. c Src activity is also elevated in transgenic mouse models of breast cancer, including the widely used polyomavirus middle-T antigen (PyVmT) model, which provides an opportunity to study the importance of c-Src in mammary tumorigenesis. However, germline c-Src deletion in mammary epithelial and stromal compartments complicates the interpretation of in vivo tumorigenesis studies as a result of severe defects in mammary gland development. We have therefore engineered a mouse strain in which deletion of c-Src can be targeted to the mammary epithelium. We demonstrate that mammary epithelial disruption of c-Src impairs proliferation and tumor progression driven by PyVmT in vivo. Whereas related kinases substitute for c-Src in PyVmT signaling, c-Src ablation impairs cell cycle progression with decreased cyclin expression and elevated expression of cyclin-dependent kinase inhibitors. Our data indicate that c-Src has essential and unique functions in proliferation and tumor progression in this mouse model that may also be important in certain contexts in some human breast cancers. PMID- 21628574 TI - Multilineage-differentiating stress-enduring (Muse) cells are a primary source of induced pluripotent stem cells in human fibroblasts. AB - The stochastic and elite models have been proposed for the mechanism of induced pluripotent stem (iPS) cell generation. In this study we report a system that supports the elite model. We previously identified multilineage-differentiating stress-enduring (Muse) cells in human dermal fibroblasts that are characterized by stress tolerance, expression of pluripotency markers, self-renewal, and the ability to differentiate into endodermal-, mesodermal-, and ectodermal-lineage cells from a single cell. They can be isolated as stage-specific embryonic antigen-3/CD105 double-positive cells. When human fibroblasts were separated into Muse and non-Muse cells and transduced with Oct3/4, Sox2, Klf4, and c-Myc, iPS cells were generated exclusively from Muse cells but not from non-Muse cells. Although some colonies were formed from non-Muse cells, they were unlike iPS cells. Furthermore, epigenetic alterations were not seen, and some of the major pluripotency markers were not expressed for the entire period during iPS cell generation. These findings were confirmed further using cells transduced with a single polycistronic virus vector encoding all four factors. The results demonstrate that in adult human fibroblasts a subset of preexisting adult stem cells whose properties are similar in some respects to those of iPS cells selectively become iPS cells, but the remaining cells make no contribution to the generation of iPS cells. Therefore this system seems to fit the elite model rather than the stochastic model. PMID- 21628575 TI - Benchmark map of forest carbon stocks in tropical regions across three continents. AB - Developing countries are required to produce robust estimates of forest carbon stocks for successful implementation of climate change mitigation policies related to reducing emissions from deforestation and degradation (REDD). Here we present a "benchmark" map of biomass carbon stocks over 2.5 billion ha of forests on three continents, encompassing all tropical forests, for the early 2000s, which will be invaluable for REDD assessments at both project and national scales. We mapped the total carbon stock in live biomass (above- and belowground), using a combination of data from 4,079 in situ inventory plots and satellite light detection and ranging (Lidar) samples of forest structure to estimate carbon storage, plus optical and microwave imagery (1-km resolution) to extrapolate over the landscape. The total biomass carbon stock of forests in the study region is estimated to be 247 Gt C, with 193 Gt C stored aboveground and 54 Gt C stored belowground in roots. Forests in Latin America, sub-Saharan Africa, and Southeast Asia accounted for 49%, 25%, and 26% of the total stock, respectively. By analyzing the errors propagated through the estimation process, uncertainty at the pixel level (100 ha) ranged from +/- 6% to +/- 53%, but was constrained at the typical project (10,000 ha) and national (>1,000,000 ha) scales at ca. +/- 5% and ca. +/- 1%, respectively. The benchmark map illustrates regional patterns and provides methodologically comparable estimates of carbon stocks for 75 developing countries where previous assessments were either poor or incomplete. PMID- 21628576 TI - Elucidating glycosaminoglycan-protein-protein interactions using carbohydrate microarray and computational approaches. AB - Glycosaminoglycan polysaccharides play critical roles in many cellular processes, ranging from viral invasion and angiogenesis to spinal cord injury. Their diverse biological activities are derived from an ability to regulate a remarkable number of proteins. However, few methods exist for the rapid identification of glycosaminoglycan-protein interactions and for studying the potential of glycosaminoglycans to assemble multimeric protein complexes. Here, we report a multidisciplinary approach that combines new carbohydrate microarray and computational modeling methodologies to elucidate glycosaminoglycan-protein interactions. The approach was validated through the study of known protein partners for heparan and chondroitin sulfate, including fibroblast growth factor 2 (FGF2) and its receptor FGFR1, the malarial protein VAR2CSA, and tumor necrosis factor-alpha (TNF-alpha). We also applied the approach to identify previously undescribed interactions between a specific sulfated epitope on chondroitin sulfate, CS-E, and the neurotrophins, a critical family of growth factors involved in the development, maintenance, and survival of the vertebrate nervous system. Our studies show for the first time that CS is capable of assembling multimeric signaling complexes and modulating neurotrophin signaling pathways. In addition, we identify a contiguous CS-E-binding site by computational modeling that suggests a potential mechanism to explain how CS may promote neurotrophin tyrosine receptor kinase (Trk) complex formation and neurotrophin signaling. Together, our combined microarray and computational modeling methodologies provide a general, facile means to identify new glycosaminoglycan-protein-protein interactions, as well as a molecular-level understanding of those complexes. PMID- 21628577 TI - Dual amyloid domains promote differential functioning of the chaplin proteins during Streptomyces aerial morphogenesis. AB - The chaplin proteins are functional amyloids found in the filamentous Streptomyces bacteria. These secreted proteins are required for the aerial development of Streptomyces coelicolor, and contribute to an intricate rodlet ultrastructure that decorates the surfaces of aerial hyphae and spores. S. coelicolor encodes eight chaplin proteins. Previous studies have revealed that only three of these proteins (ChpC, ChpE, and ChpH) are necessary for promoting aerial development, and of these three, ChpH is the primary developmental determinant. Here, we show that the model chaplin, ChpH, contains two amyloidogenic domains: one in the N terminus and one in the C terminus of the mature protein. These domains have different polymerization properties as determined using fluorescence spectroscopy, secondary structure analyses, and electron microscopy. We coupled these in vitro assays with in vivo genetic studies to probe the connection between ChpH amyloidogenesis and its biological function. Using mutational analyses, we demonstrated that both N- and C-terminal amyloid domains of ChpH were required for promoting aerial hypha formation, while the N-terminal domain was dispensable for assembly of the rodlet ultrastructure. These results suggest that there is a functional differentiation of the dual amyloid domains in the chaplin proteins. PMID- 21628578 TI - Recruitment interactions can override catalytic interactions in determining the functional identity of a protein kinase. AB - The yeast Saccharomyces cerevisae has four distinct mitogen-activated protein kinase kinases (MAPKKs), each of which has a distinct functional identity characterized by communication with specific upstream and downstream partners to form distinct functional pathways. These four kinases belong to one family, sharing closely related catalytic domains. How have these four related kinases diverged to take on four distinct functional roles? The specificity of an enzyme for a particular substrate is often thought to reside in differences in the catalytic domain. However, many kinases, including MAPKKs, have modular interaction domains and motifs that have been shown to play an important role in determining the specificity of kinases through recruitment to specific partners and complexes. Here we probe the relative importance of catalytic domain interactions versus recruitment interactions in defining the functional identity of MAPKKs by asking whether we can use recruitment interactions to force other MAPKK catalytic domains to play the functional role of the mating MAPKK, Ste7. We find that two alternative MAPKKs, Pbs2 and Mkk2, can be forced to functionally replace the mating MAPKK Ste7, but only if the proper set of recruitment interactions are grafted onto their catalytic domains. These results show that within a family of kinases, recruitment interactions can play a dominant role in defining functional identity, and is consistent with a model in which new kinase functions can arise through recombination of existing catalytic domains with new interaction modules. PMID- 21628579 TI - Clofarabine 5'-di and -triphosphates inhibit human ribonucleotide reductase by altering the quaternary structure of its large subunit. AB - Human ribonucleotide reductases (hRNRs) catalyze the conversion of nucleotides to deoxynucleotides and are composed of alpha- and beta-subunits that form active alpha(n)beta(m) (n, m = 2 or 6) complexes. alpha binds NDP substrates (CDP, UDP, ADP, and GDP, C site) as well as ATP and dNTPs (dATP, dGTP, TTP) allosteric effectors that control enzyme activity (A site) and substrate specificity (S site). Clofarabine (ClF), an adenosine analog, is used in the treatment of refractory leukemias. Its mode of cytotoxicity is thought to be associated in part with the triphosphate functioning as an allosteric inhibitor of hRNR. Studies on the mechanism of inhibition of hRNR by ClF di- and triphosphates (ClFDP and ClFTP) are presented. ClFTP is a reversible inhibitor (K(i) = 40 nM) that rapidly inactivates hRNR. However, with time, 50% of the activity is recovered. D57N-alpha, a mutant with an altered A site, prevents inhibition by ClFTP, suggesting its A site binding. ClFDP is a slow-binding, reversible inhibitor ( K(i)*; t(1/2) = 23 min). CDP protects alpha from its inhibition. The altered off-rate of ClFDP from E*ClFDP* by ClFTP (A site) or dGTP (S site) and its inhibition of D57N-alpha together implicate its C site binding. Size exclusion chromatography of hRNR or alpha alone with ClFDP or ClFTP, +/- ATP or dGTP, reveals in each case that alpha forms a kinetically stable hexameric state. This is the first example of hexamerization of alpha induced by an NDP analog that reversibly binds at the active site. PMID- 21628580 TI - Enhanced chemistry-climate feedbacks in past greenhouse worlds. AB - Trace greenhouse gases are a fundamentally important component of Earth's global climate system sensitive to global change. However, their concentration in the pre-Pleistocene atmosphere during past warm greenhouse climates is highly uncertain because we lack suitable geochemical or biological proxies. This long standing issue hinders assessment of their contribution to past global warmth and the equilibrium climate sensitivity of the Earth system (E(ss)) to CO(2). Here we report results from a series of three-dimensional Earth system modeling simulations indicating that the greenhouse worlds of the early Eocene (55 Ma) and late Cretaceous (90 Ma) maintained high concentrations of methane, tropospheric ozone, and nitrous oxide. Modeled methane concentrations were four- to fivefold higher than the preindustrial value typically adopted in modeling investigations of these intervals, even after accounting for the possible high CO(2)-suppression of biogenic isoprene emissions on hydroxyl radical abundance. Higher concentrations of trace greenhouse gases exerted marked planetary heating (> 2 K), amplified in the high latitudes (> 6 K) by lower surface albedo feedbacks, and increased E(ss) in the Eocene by 1 K. Our analyses indicate the requirement for including non-CO(2) greenhouse gases in model-based E(ss) estimates for comparison with empirical paleoclimate assessments, and point to chemistry climate feedbacks as possible amplifiers of climate sensitivity in the Anthropocene. PMID- 21628581 TI - Visualization of the nanospring dynamics of the IkappaBalpha ankyrin repeat domain in real time. AB - IkappaBalpha is a crucial regulator of NFkappaB transcription. NFkappaB-mediated gene activation is robust because levels of free IkappaBalpha are kept extremely low by rapid, ubiquitin-independent degradation of newly synthesized IkappaBalpha. IkappaBalpha has a weakly folded ankyrin repeat 5-6 (AR5-6) region that is critical in establishing its short intracellular half-life. The AR5-6 region of IkappaBalpha folds upon binding to NFkappaB. The NFkappaB-bound IkappaBalpha has a long half-life and requires ubiquitin-targeted degradation. We present single molecule FRET evidence that the native state of IkappaBalpha transiently populates an intrinsically disordered state characterized by a more extended structure and fluctuations on the millisecond time scale. Binding to NFkappaB or introduction of stabilizing mutations in AR 6 suppressed the fluctuations, whereas higher temperature or small amounts of urea increased them. The results reveal that intrinsically disordered protein regions transition between collapsed and extended conformations under native conditions. PMID- 21628582 TI - One-dimensional polaritons with size-tunable and enhanced coupling strengths in semiconductor nanowires. AB - Strong coupling of light with excitons in direct bandgap semiconductors leads to the formation of composite photonic-electronic quasi-particles (polaritons), in which energy oscillates coherently between the photonic and excitonic states with the vacuum Rabi frequency. The light-matter coherence is maintained until the oscillator dephases or the photon escapes. Exciton-polariton formation has enabled the observation of Bose-Einstein condensation in the solid-state, low threshold polariton lasing and is also useful for terahertz and slow-light applications. However, maintaining coherence for higher carrier concentration and temperature applications still requires increased coupling strengths. Here, we report on size-tunable, exceptionally high exciton-polariton coupling strengths characterized by a vacuum Rabi splitting of up to 200 meV as well as a reduction in group velocity, in surface-passivated, self-assembled semiconductor nanowire cavities. These experiments represent systematic investigations on light-matter coupling in one-dimensional optical nanocavities, demonstrating the ability to engineer light-matter coupling strengths at the nanoscale, even in non-quantum confined systems, to values much higher than in bulk. PMID- 21628583 TI - Unusual regioversatility of acetyltransferase Eis, a cause of drug resistance in XDR-TB. AB - The emergence of multidrug-resistant and extensively drug-resistant (XDR) tuberculosis (TB) is a serious global threat. Aminoglycoside antibiotics are used as a last resort to treat XDR-TB. Resistance to the aminoglycoside kanamycin is a hallmark of XDR-TB. Here, we reveal the function and structure of the mycobacterial protein Eis responsible for resistance to kanamycin in a significant fraction of kanamycin-resistant Mycobacterium tuberculosis clinical isolates. We demonstrate that Eis has an unprecedented ability to acetylate multiple amines of many aminoglycosides. Structural and mutagenesis studies of Eis indicate that its acetylation mechanism is enabled by a complex tripartite fold that includes two general control non-derepressible 5 (GCN5)-related N acetyltransferase regions. An intricate negatively charged substrate-binding pocket of Eis is a potential target of new antitubercular drugs expected to overcome aminoglycoside resistance. PMID- 21628584 TI - Structural plasticity of a transmembrane peptide allows self-assembly into biologically active nanoparticles. AB - Significant efforts have been devoted to the development of nanoparticular delivering systems targeting tumors. However, clinical application of nanoparticles is hampered by insufficient size homogeneity, difficulties in reproducible synthesis and manufacturing, frequent high uptake in the liver, systemic toxicity of the carriers (particularly for inorganic nanoparticles), and insufficient selectivity for tumor cells. We have found that properly modified synthetic analogs of transmembrane domains of membrane proteins can self-assemble into remarkably uniform spherical nanoparticles with innate biological activity. Self-assembly is driven by a structural transition of the peptide that adopts predominantly a beta-hairpin conformation in aqueous solutions, but folds into an alpha-helix upon spontaneous fusion of the nanoparticles with cell membrane. A 24 amino acid peptide corresponding to the second transmembrane helix of the CXCR4 forms self-assembled particles that inhibit CXCR4 function in vitro and hamper CXCR4-dependent tumor metastasis in vivo. Furthermore, such nanoparticles can encapsulate hydrophobic drugs, thus providing a delivery system with the potential for dual biological activity. PMID- 21628585 TI - Structure of transcription factor HetR required for heterocyst differentiation in cyanobacteria. AB - HetR is an essential regulator of heterocyst development in cyanobacteria. HetR binds to a DNA palindrome upstream of the hetP gene. We report the crystal structure of HetR from Fischerella at 3.0 A. The protein is a dimer comprised of a central DNA-binding unit containing the N-terminal regions of the two subunits organized with two helix-turn-helix motifs; two globular flaps extending in opposite directions; and a hood over the central core formed from the C-terminal subdomains. The flaps and hood have no structural precedent in the protein database, therefore representing new folds. The structural assignments are supported by site-directed mutagenesis and DNA-binding studies. We suggest that HetR serves as a scaffold for assembly of transcription components critical for heterocyst development. PMID- 21628586 TI - Abrupt Holocene climate change as an important factor for human migration in West Greenland. AB - West Greenland has had multiple episodes of human colonization and cultural transitions over the past 4,500 y. However, the explanations for these large scale human migrations are varied, including climatic factors, resistance to adaptation, economic marginalization, mercantile exploration, and hostile neighborhood interactions. Evaluating the potential role of climate change is complicated by the lack of quantitative paleoclimate reconstructions near settlement areas and by the relative stability of Holocene temperature derived from ice cores atop the Greenland ice sheet. Here we present high-resolution records of temperature over the past 5,600 y based on alkenone unsaturation in sediments of two lakes in West Greenland. We find that major temperature changes in the past 4,500 y occurred abruptly (within decades), and were coeval in timing with the archaeological records of settlement and abandonment of the Saqqaq, Dorset, and Norse cultures, which suggests that abrupt temperature changes profoundly impacted human civilization in the region. Temperature variations in West Greenland display an antiphased relationship to temperature changes in Ireland over centennial to millennial timescales, resembling the interannual to multidecadal temperature seesaw associated with the North Atlantic Oscillation. PMID- 21628587 TI - Policy perils of ignoring uncertainty in oil palm research. PMID- 21628588 TI - A program of microRNAs controls osteogenic lineage progression by targeting transcription factor Runx2. AB - Lineage progression in osteoblasts and chondrocytes is stringently controlled by the cell-fate-determining transcription factor Runx2. In this study, we directly addressed whether microRNAs (miRNAs) can control the osteogenic activity of Runx2 and affect osteoblast maturation. A panel of 11 Runx2-targeting miRNAs (miR-23a, miR-30c, miR-34c, miR-133a, miR-135a, miR-137, miR-204, miR-205, miR-217, miR 218, and miR-338) is expressed in a lineage-related pattern in mesenchymal cell types. During both osteogenic and chondrogenic differentiation, these miRNAs, in general, are inversely expressed relative to Runx2. Based on 3'UTR luciferase reporter, immunoblot, and mRNA stability assays, each miRNA directly attenuates Runx2 protein accumulation. Runx2-targeting miRNAs differentially inhibit Runx2 protein expression in osteoblasts and chondrocytes and display different efficacies. Thus, cellular context contributes to miRNA-mediated regulation of Runx2. All Runx2-targeting miRNAs (except miR-218) significantly impede osteoblast differentiation, and their effects can be reversed by the corresponding anti-miRNAs. These findings demonstrate that osteoblastogenesis is limited by an elaborate network of functionally tested miRNAs that directly target the osteogenic master regulator Runx2. PMID- 21628589 TI - Minimal requirements for actin filament disassembly revealed by structural analysis of malaria parasite actin-depolymerizing factor 1. AB - Malaria parasite cell motility is a process that is dependent on the dynamic turnover of parasite-derived actin filaments. Despite its central role, actin's polymerization state is controlled by a set of identifiable regulators that is markedly reduced compared with those of other eukaryotic cells. In Plasmodium falciparum, the most virulent species that affects humans, this minimal repertoire includes two members of the actin-depolymerizing factor/cofilin (AC) family of proteins, P. falciparum actin-depolymerizing factor 1 (PfADF1) and P. falciparum actin-depolymerizing factor 2. This essential class of actin regulator is involved in the control of filament dynamics at multiple levels, from monomer binding through to filament depolymerization and severing. Previous biochemical analyses have suggested that PfADF1 sequesters monomeric actin but, unlike most eukaryotic counterparts, has limited potential to bind or depolymerize filaments. The molecular basis for these unusual properties and implications for parasite cell motility have not been established. Here we present the crystal structure of an apicomplexan AC protein, PfADF1. We show that PfADF1 lacks critical residues previously implicated as essential for AC-mediated actin filament binding and disassembly, having a substantially reduced filament-binding loop and C-terminal alpha4 helix. Despite this divergence in structure, we demonstrate that PfADF1 is capable of efficient actin filament severing. Furthermore, this severing occurs despite PfADF1's low binding affinity for filaments. Comparative structural analysis along with biochemical and microscopy evidence establishes that severing is reliant on the availability of an exposed basic residue in the filament binding loop, a conserved minimal requirement that defines AC-mediated filament disassembly across eukaryotic cells. PMID- 21628590 TI - Proliferating cell nuclear antigen (PCNA)-associated KIAA0101/PAF15 protein is a cell cycle-regulated anaphase-promoting complex/cyclosome substrate. AB - The anaphase-promoting complex/cyclosome (APC/C) is a cell cycle-regulated E3 ubiquitin ligase that controls the degradation of substrate proteins at mitotic exit and throughout the G1 phase. We have identified an APC/C substrate and cell cycle-regulated protein, KIAA0101/PAF15. PAF15 protein levels peak in the G2/M phase of the cell cycle and drop rapidly at mitotic exit in an APC/C- and KEN-box dependent fashion. PAF15 associates with proliferating cell nuclear antigen (PCNA), and depletion of PAF15 decreases the number of cells in S phase, suggesting a role for it in cell cycle regulation. Following irradiation, PAF15 colocalized with gammaH2AX foci at sites of DNA damage through its interaction with PCNA. Finally, PAF15 depletion led to an increase in homologous recombination-mediated DNA repair, and overexpression caused sensitivity to UV induced DNA damage. We conclude that PAF15 is an APC/C-regulated protein involved in both cell cycle progression and the DNA damage response. PMID- 21628591 TI - Retraction for Nemecek et al.: A prion of yeast metacaspase homolog (Mca1p) detected by a genetic screen. PMID- 21628592 TI - In vivo diagnosis of murine pancreatic intraepithelial neoplasia and early-stage pancreatic cancer by molecular imaging. AB - Pancreatic ductal adenocarcinoma (PDAC) is a fatal disease with poor patient outcome often resulting from late diagnosis in advanced stages. To date methods to diagnose early-stage PDAC are limited and in vivo detection of pancreatic intraepithelial neoplasia (PanIN), a preinvasive precursor of PDAC, is impossible. Using a cathepsin-activatable near-infrared probe in combination with flexible confocal fluorescence lasermicroscopy (CFL) in a genetically defined mouse model of PDAC we were able to detect and grade murine PanIN lesions in real time in vivo. Our diagnostic approach is highly sensitive and specific and proved superior to clinically established fluorescein-enhanced imaging. Translation of this endoscopic technique into the clinic should tremendously improve detection of pancreatic neoplasia, thus reforming management of patients at risk for PDAC. PMID- 21628593 TI - Neonatal Fc receptor for IgG (FcRn) regulates cross-presentation of IgG immune complexes by CD8-CD11b+ dendritic cells. AB - Cross-presentation of IgG-containing immune complexes (ICs) is an important means by which dendritic cells (DCs) activate CD8(+) T cells, yet it proceeds by an incompletely understood mechanism. We show that monocyte-derived CD8(-)CD11b(+) DCs require the neonatal Fc receptor for IgG (FcRn) to conduct cross-presentation of IgG ICs. Consequently, in the absence of FcRn, Fcgamma receptor (FcgammaR) mediated antigen uptake fails to initiate cross-presentation. FcRn is shown to regulate the intracellular sorting of IgG ICs to the proper destination for such cross-presentation to occur. We demonstrate that FcRn traps antigen and protects it from degradation within an acidic loading compartment in association with the rapid recruitment of key components of the phagosome-to-cytosol cross presentation machinery. This unique mechanism thus enables cross-presentation to evolve from an atypically acidic loading compartment. FcRn-driven cross presentation is further shown to control cross-priming of CD8(+) T-cell responses in vivo such that during chronic inflammation, FcRn deficiency results in inadequate induction of CD8(+) T cells. These studies thus demonstrate that cross presentation in CD8(-)CD11b(+) DCs requires a two-step mechanism that involves FcgammaR-mediated internalization and FcRn-directed intracellular sorting of IgG ICs. Given the centrality of FcRn in controlling cross-presentation, these studies lay the foundation for a unique means to therapeutically manipulate CD8(+) T-cell responses. PMID- 21628594 TI - The trans-Himalayan flights of bar-headed geese (Anser indicus). AB - Birds that fly over mountain barriers must be capable of meeting the increased energetic cost of climbing in low-density air, even though less oxygen may be available to support their metabolism. This challenge is magnified by the reduction in maximum sustained climbing rates in large birds. Bar-headed geese (Anser indicus) make one of the highest and most iconic transmountain migrations in the world. We show that those populations of geese that winter at sea level in India are capable of passing over the Himalayas in 1 d, typically climbing between 4,000 and 6,000 m in 7-8 h. Surprisingly, these birds do not rely on the assistance of upslope tailwinds that usually occur during the day and can support minimum climb rates of 0.8-2.2 km.h(-1), even in the relative stillness of the night. They appear to strategically avoid higher speed winds during the afternoon, thus maximizing safety and control during flight. It would seem, therefore, that bar-headed geese are capable of sustained climbing flight over the passes of the Himalaya under their own aerobic power. PMID- 21628595 TI - Profile of Charles L. Sawyers. PMID- 21628596 TI - Drosophila homeodomain-interacting protein kinase inhibits the Skp1-Cul1-F-box E3 ligase complex to dually promote Wingless and Hedgehog signaling. AB - Drosophila Homeodomain-interacting protein kinase (Hipk) has been shown to regulate in vivo, the stability of Armadillo, the transcriptional effector of Wingless signaling. The Wingless pathway culminates in the stabilization of Armadillo that, in the absence of signaling, is sequentially phosphorylated, polyubiquitinated and degraded. Loss-of-function clones for hipk result in reduced stabilized Armadillo, whereas overexpression of hipk elevates Armadillo levels to promote Wingless-responsive target gene expression. Here, we show that overexpression of hipk can suppress the effects of negative regulators of Armadillo to prevent its degradation in the wing imaginal disc. Hipk acts to stabilize Armadillo by impeding the function of the E3 ubiquitin ligase Skp1-Cul1 F-box (SCF)(Slimb), thereby inhibiting Armadillo ubiquitination and subsequent degradation. Vertebrate Hipk2 displays a similar ability to prevent beta-catenin ubiquitination in a functionally conserved mechanism. We find that Hipk's ability to inhibit SCF(Slimb)-mediated ubiquitination is not restricted to Armadillo and extends to other substrates of SCF(Slimb), including the Hedgehog signaling effector Ci. Thus, similar to casein kinase 1 and glycogen synthase kinase 3, Hipk dually regulates both Wingless and Hedgehog signaling by controlling the stability of their respective signaling effectors, but it is the first kinase to our knowledge identified that promotes the stability of both Armadillo and Ci. PMID- 21628597 TI - Protein-specific manipulation of ejaculate composition in response to female mating status in Drosophila melanogaster. AB - Female promiscuity can generate postcopulatory competition among males, but it also provides the opportunity for exploitation of rival male ejaculates. For example, in many insect species, male seminal fluid proteins (Sfps) transferred in a female's first mating stimulate increased fecundity and decreased receptivity to remating. Subsequent mates of females could potentially take advantage of the effects of the first male's Sfps and strategically reduce investment in their own ejaculate. We compared postmating responses (fecundity and sexual receptivity) of Drosophila melanogaster females after their first (virgin) matings (V), to the responses of females remating (M) 24 h after their first mating. The results show that M matings fail to boost fecundity and, thus, males are unlikely to gain fitness from transferring Sfps whose sole function-in V matings-is fecundity-stimulation. However, males can protect their likelihood of paternity in M matings through the transfer of receptivity-inhibiting Sfps. The levels of a fecundity-stimulating Sfp (ovulin) were significantly lower in M females relative to V females, at the same time point shortly after the end of mating. In contrast, the levels of a key receptivity-inhibiting Sfp (sex peptide) were the same in M and V females. These results support the hypothesis that males can adaptively tailor the composition of proteins in the ejaculate, allowing a male to take advantage of the fecundity-stimulating effects of the previous male's ovulin, yet maintaining investment in sex peptide. Furthermore, our results demonstrate sophisticated protein-specific ejaculate manipulation. PMID- 21628598 TI - Racial identification, racial discrimination, and substance use vulnerability among African American young adults. AB - Two studies examined racial identity (RI) as a protective factor against substance use cognitions among African American young adults who either envisioned or experienced racial discrimination. In Study 1, participants envisioned a discrimination or nondiscrimination scenario, and then their willingness to use drugs and an indirect measure of substance use were assessed. Discrimination was associated with higher levels of use cognitions among participants with low levels of RI. In Study 2, participants were excluded or included in an online game (Cyberball) by White peers and then engaged in an RI affirmation or control writing task. Participants attributed this exclusion to racial discrimination. Excluded participants who did not affirm their RI reported the highest levels of substance use cognitions, especially if they had engaged in higher levels of previous substance use. These findings highlight the importance of RI among Black young adults and the impact of discrimination on health behaviors. PMID- 21628599 TI - New insights: dose-response relationship between psychotropic drugs and falls: a study in nursing home residents with dementia. AB - The contribution of specific psychotropic drugs to fall risk in patients with dementia has not been quantified precisely until now. The authors evaluated the dose-response relationship between psychotropic drugs and falls in nursing home residents with dementia. Daily drug use and daily falls were recorded in 248 nursing home residents with dementia from January 1, 2006, to January 1, 2008. For each day of the study period, data on drug use were abstracted from the prescription database, and falls were retrieved from a standardized incident report system, resulting in a data set of 85 074 person-days. The authors found significant dose-response relationships for the use of antipsychotics (hazard ratio [HR], 2.78; 95% confidence interval [CI], 1.49-5.17), anxiolytics (1.60; 1.20-2.14), hypnotics and sedatives (2.58; 1.42-4.68), and antidepressants (2.84; 1.93-4.16). Fall risk increased significantly with 28% at 0.25 of the defined daily dose (DDD) of an antipsychotic or antidepressant, with 8% at 0.2 of the DDD of an anxiolytic, and with 56% at 0.5 of the DDD of a hypnotic or sedative; it increased further with dose increments and with combinations of psychotropics. Even at low dosages, psychotropic drugs are associated with increased fall risk in nursing home residents with dementia. PMID- 21628600 TI - The monoamine oxidase type B inhibitor rasagiline in the treatment of Parkinson disease: is tyramine a challenge? AB - Rasagiline is an irreversible monoamine oxidase type B (MAO-B) inhibitor indicated for the treatment of the signs and symptoms of idiopathic Parkinson disease as initial monotherapy and as adjunct therapy to levodopa. Pharmacologic inhibition of monoamine oxidase type A (MAO-A), but not MAO-B, poses a risk of the "cheese effect," a hypertensive response to excess dietary tyramine, a biogenic sympathomimetic amine. Tyramine challenge studies, conducted to characterize rasagiline selectivity for the MAO-B enzyme and tyramine sensitivity, demonstrate that rasagiline, when used at the recommended dose, is selective for MAO-B and is not associated with heightened tyramine sensitivity. This conclusion is also supported by safety results from large clinical trials of rasagiline in Parkinson disease involving 2066 rasagiline-treated patients who did not require dietary tyramine restriction per protocol. In late 2009, US labeling for rasagiline was modified to state that dietary tyramine restrictions are not ordinarily required when rasagiline is administered at recommended doses. In addition, because rasagiline has been demonstrated to be selective for MAO-B at the approved dose of up to 1 mg/d, contraindications regarding concomitant use with sympathomimetic amines, use of sympathomimetic vasopressors in conjunction with general or local anesthesia, and use in patients with pheochromocytoma also were removed. PMID- 21628601 TI - Relationship between exposure to prasugrel active metabolite and clinical outcomes in the TRITON-TIMI 38 substudy. AB - In TRITON-TIMI 38, levels of the prasugrel active metabolite (pras-AM) were measured in a population pharmacokinetic substudy that characterized the intrinsic and extrinsic factors influencing exposure. Higher exposure to the pras AM was observed in low-weight or very elderly patients. The authors hypothesized that this higher exposure might explain the higher risk of non-coronary artery bypass graft (CABG)-related TIMI-related bleeding observed in these 2 patient populations. The relationship between estimated exposure to the pras-AM and clinical outcomes was assessed in 1159 prasugrel-treated patients enrolled in the substudy using multivariable logistic regression analysis. There was no relationship between pras-AM exposure and efficacy through 3 days or after 3 days. Higher estimated pras-AM exposure was associated with a higher incidence of non-CABG-related TIMI major or minor bleeding after 3 days (P < .05) but not through 3 days from start of study drug. Factors associated with increased risk for non-CABG-related TIMI bleeding (>=75 years and <60 kg) also identified subgroups with higher exposure to the pras-AM. Within low body weight and very elderly subgroups, bleeding was largely confined to patients having the highest exposure to the pras-AM, indicating that a prasugrel lower dose in these subgroups may reduce the risk of bleeding while maintaining efficacy. PMID- 21628602 TI - Pharmacokinetics and relative bioavailability of a fixed-dose combination of enteric-coated naproxen and non-enteric-coated esomeprazole magnesium. AB - This randomized, 4-way crossover study assessed the single-dose pharmacokinetics and relative bioavailability of naproxen and esomeprazole after administration of a fixed-dose combination tablet of enteric-coated (EC) naproxen 500 mg and non-EC esomeprazole magnesium 20 mg (NAP/ESO tablet). Equivalent doses of an EC naproxen tablet plus an EC esomeprazole magnesium capsule taken concomitantly, an EC naproxen tablet alone, or an EC esomeprazole magnesium capsule alone were used as comparators. Forty healthy adults were randomized to receive 4 study treatments with a washout interval >=12 days. Naproxen plasma profiles were similar between the NAP/ESO tablet and EC naproxen, although median t(max) was longest for the NAP/ESO tablet (5.3 vs 3.5-4.0 hours). Naproxen C(max), AUC(0-infinity), and AUC(0-t) showed bioequivalence between naproxen formulations. The NAP/ESO tablet produced much shorter esomeprazole t(max) than the EC esomeprazole formulation (0.45 vs 2.5 hours). Esomeprazole C(max) and AUCs were comparable between the EC esomeprazole formulation administered with or without EC naproxen but were lower with the NAP/ESO tablet. In conclusion, there are no pharmacokinetic drug interactions between naproxen and esomeprazole. The NAP/ESO tablet is bioequivalent to EC naproxen, and as expected, the bioavailability of non-EC esomeprazole from the NAP/ESO tablet is lower than the EC esomeprazole formulation. PMID- 21628603 TI - Safety and tolerability of aclidinium administered intravenously and absolute bioavailability of inhaled aclidinium in healthy male participants. AB - Aclidinium bromide is a long-acting muscarinic antagonist in development for chronic obstructive pulmonary disease treatment. This 2-part, phase I study evaluated the safety and tolerability of single ascending intravenous (IV) doses of aclidinium to determine its maximum tolerated dose (MTD; part I) and its absolute bioavailability (part II). Healthy male participants (N = 24) were randomized (1:1) in each part: 3-period crossover, placebo-controlled, single ascending, alternating IV doses of aclidinium (25-400 ug) in part I and 2-period crossover, single-alternating IV and inhaled doses of aclidinium (200 ug) in part II. A >=7-day washout separated treatment periods. Pharmacokinetic data were collected in both parts. Following IV or inhaled aclidinium, time to reach maximum plasma concentration following drug administration (t(max) ) was 5 to 7 minutes for all doses. After maximum plasma drug concentration (C(max)), aclidinium was rapidly cleared from plasma. Aclidinium absolute bioavailability was <5% following a single inhaled 200-ug dose. Urinary excretion of unchanged aclidinium was very low, with a greater amount of inactive metabolites excreted compared with aclidinium, all of which were recovered within 12 hours postdose. The MTD following IV administration was not reached; all single IV (25-400 ug) and inhaled doses (200 ug) were well tolerated. In conclusion, the low and short lived bioavailability of aclidinium and the low incidence of systemic side effects contribute to its positive safety and tolerability profile. PMID- 21628604 TI - Valproate reduces the glucuronidation of asenapine without affecting asenapine plasma concentrations. AB - Asenapine is indicated for treatment of schizophrenia in the United States and acute treatment of manic or mixed episodes, as monotherapy (United States and European Union) or adjunct therapy (United States only), associated with bipolar I disorder. It is extensively metabolized; the 2 main metabolites are asenapine N glucuronide and N-desmethyl-asenapine. The authors investigated the pharmacokinetic interactions between asenapine and valproate in an open-label, randomized, 2-way crossover study. Twenty-four healthy male volunteers received sublingual doses of asenapine 5 mg alone or under steady-state valproate (500 mg bid for 9 days). Blood samples collected until 72 hours postdosing were analyzed for asenapine, N-desmethyl-asenapine, and asenapine N-glucuronide. Compared with asenapine alone, valproate substantially reduced N-glucuronide formation (area under the curve from 0 to infinity [AUC(0-infinity)] reduced 7.4-fold, maximum concentration [C(max)] reduced 6.6-fold) and moderately reduced N-desmethyl asenapine formation (AUC(0-infinity) reduced 30%, C(max) unchanged). Coadministration of valproate did not affect asenapine AUC(0-infinity) and C(max) (confidence intervals for the ratios of asenapine AUC(0-infinity) and C(max) were contained within the predefined 0.80-1.25 acceptance range). Low-dose valproate, although almost completely inhibiting glucuronidation of asenapine, did not affect the pharmacokinetics of asenapine itself, the entity primarily responsible for the pharmacologic effects of the drug. PMID- 21628605 TI - Accelerometer-triggered electrical stimulation for reach and grasp in chronic stroke patients: a pilot study. AB - BACKGROUND: Electrical stimulation of the upper extremity may reduce impairment in patients following stroke. Stimulation triggered on demand combined with task practice may be an effective means of promoting recovery of function. OBJECTIVE: The authors investigated the feasibility of using accelerometer-controlled electrical stimulation for the elbow, wrist, and finger extensors to enable functional task practice in patients with chronic hemiparesis. METHODS: Following a 4-week baseline, participants received 2 weeks of cyclic stimulation exercise to elbow and forearm extensor muscles, followed by 10 weeks of triggered stimulation to practice functional reaching. Participants were reassessed 12 weeks later as well. Outcome measures were the Action Research Arm Test (ARAT), Modified Ashworth Scale (MAS), Canadian Occupational Performance Measure (COPM), Psychosocial Impact of Assistive Devices Scale (PIADS), and Use of Device Questionnaire (UDQ). RESULTS: Fifteen volunteers who had at least 45 degrees of forward shoulder flexion and could initiate elbow extension and grasp completed the study. The ARAT score improved from 19 to 32 (P = .002); the MAS score for elbow, wrist, and finger flexor spasticity was reduced from 2 each to 1, 0, and 1 (P < .05); the COPM performance and satisfaction scores improved (P = .001); and the PIADS became positive for competence (P = .005), adaptability (P = .008), and self-esteem (P = .008). Gains were maintained 12 weeks later. CONCLUSIONS: Accelerometer-triggered electrical stimulation to augment task training for the hemiplegic arm is feasible and may improve functional ability and quality of life which may be maintained 12 weeks after treatment. A randomized trial design is required to evaluate efficacy and cost benefit. PMID- 21628606 TI - An evidence-based review of a Lentinula edodes mushroom extract as complementary therapy in the surgical oncology patient. AB - The purpose of this review is to present the currently published evidence regarding the use, efficacy, potential mechanisms of action, and results of published clinical trials regarding the use of a Lentinula edodes mushroom derived extract (active hexose correlated compound) as complementary therapy in patients with cancer. The authors explore the current preclinical and clinical evidence as it relates to this topic and its potential use in the surgical oncology patient. There has been a growing interest in stimulation of the immune system in trauma, cancer, and surgical patients in general. Little, however, has been written about some-of the supplements in widely used in Japan and China, but relatively unheard of in the United States. PMID- 21628607 TI - Early versus traditional postoperative feeding in patients undergoing resectional gastrointestinal surgery: a meta-analysis. AB - BACKGROUND: A meta-analysis evaluating surgical outcomes following nutritional provision provided proximal to the anastomosis within 24 hours of gastrointestinal surgery compared with traditional postoperative management was conducted. METHODS: Databases were searched to identify randomized controlled trials comparing the outcomes of early and traditional postoperative feeding. Trials involving gastrointestinal tract resection followed by patients receiving nutritionally significant oral or enteral intake within 24 hours after surgery were included for analysis. RESULTS: Fifteen studies involving a total of 1240 patients were analyzed. A statistically significant reduction (45%) in relative odds of total postoperative complications was seen in patients receiving early postoperative feeding (odds ratio [OR] 0.55; confidence interval [CI], 0.35 0.87, P = .01). No effect of early feeding was seen with relation to anastomotic dehiscence (OR 0.75; CI, 0.39-1.4, P = .39), mortality (OR 0.71; CI, 0.32-1.56, P = .39), days to passage of flatus (weighted mean difference [WMD] -0.42; CI, 1.12 to 0.28, P = .23), first bowel motion (WMD -0.28; CI, -1.20 to 0.64, P = .55), or reduced length of stay (WMD -1.28; CI, -2.94 to 0.38, P = .13); however, the direction of clinical outcomes favored early feeding. Nasogastric tube reinsertion was less common in traditional feeding interventions (OR 1.48; CI, 0.93-2.35, P = .10). CONCLUSIONS: Early postoperative nutrition is associated with significant reductions in total complications compared with traditional postoperative feeding practices and does not negatively affect outcomes such as mortality, anastomotic dehiscence, resumption of bowel function, or hospital length of stay. PMID- 21628608 TI - Serum vitamins in adult patients with short bowel syndrome receiving intermittent parenteral nutrition. AB - BACKGROUND: Short bowel syndrome (SBS) occurs after massive intestinal resection, and parenteral nutrition (PN) therapy may be necessary even after a period of adaptation. The purpose of this study was to determine the vitamin status in adults with SBS receiving intermittent PN. METHODS: The study was conducted on hospitalized adults with SBS who were receiving intermittent PN therapy (n = 8). Nine healthy volunteers, paired by age and sex, served as controls. Food ingestion, anthropometry, plasma folic acid, and vitamins B(12), C, A, D, E, and K were evaluated. RESULTS: The levels of vitamins A, D, and B(12) in both groups were similar. SBS patients presented higher values of folic acid (21.3 +/- 4.4 vs 14.4 +/- 5.2, P = .01) and lower values of vitamin C (0.9 +/- 0.4 vs 1.2 +/- 0.3 mg/dL, P = .03), alpha-tocopherol (16.3 +/- 3.4 vs 24.1 +/- 2.7 umol/L, P < .001), and phylloquinone (0.6 +/- 0.2 vs 1.0 +/- 0.5 nmol/L, P < .03). Eight seven percent of patients had vitamin D deficiency, and all patients presented with serum vitamin E levels below reference values. CONCLUSIONS: Despite all efforts to offer all the nutrients mentioned above, SBS patients had lower serum levels of vitamins C, E, and K, similar to those observed in patients on home PN. These findings suggest that the administered vitamins were not sufficient for the intermittent PN scheme and that individual adjustments are needed depending on the patient's vitamin status. PMID- 21628609 TI - Parenteral nutrition-associated liver disease--watch the fat! PMID- 21628610 TI - Prospective study of clinical and histological safety of pure and uncontaminated Canadian oats in the management of celiac disease. AB - BACKGROUND: Pure oats are safe for most patients with celiac disease, but concerns regarding contamination by other grains limit their consumption. The Canadian Celiac Association recently released guidelines governing the production of pure oats. The objective was to test the safety of a product manufactured under these guidelines. METHODS: Fifteen adults with established, biopsy confirmed celiac disease of >= 1 year duration were challenged with 350 g/wk of pure oats for 12 weeks. Symptom scores, weight, hemoglobin, ferritin, albumin, and tissue transglutaminase (tTG) were assessed at weeks 0, 6, and 12. Duodenal biopsies were obtained before and after oat challenge and assessed based on the modified Marsh-Oberhuber score. Compliance with a gluten-free diet was monitored with random food diaries. RESULTS: Fifteen patients completed the study and were analyzed in intention-to-treat and per-protocol analyses. There were no significant changes in symptom scores, weight, hemoglobin, ferritin, or albumin during oat consumption. The tTG remained negative in all patients, and the histology scores did not significantly change during oat challenge. The only relapse occurred in a patient who became noncompliant with her gluten-free diet. CONCLUSION: The findings support the safety of pure, uncontaminated oats manufactured under Canadian Celiac Association guidelines for patients with celiac disease. PMID- 21628611 TI - 56-year-old woman with positional headache. PMID- 21628612 TI - Cystitis treatment in women, circa 2011: new role for an old drug. PMID- 21628613 TI - Colonic polyposis and neoplasia in Cowden syndrome. AB - OBJECTIVE: To identify and describe the frequency, histologic features, and clinical outcome of colon polyposis and neoplasia in Cowden syndrome--a rare familial hamartoma tumor syndrome associated with mutations in the PTEN gene. PATIENTS AND METHODS: Patients with a clinical diagnosis of PTEN hamartoma tumor syndrome-Cowden phenotype were retrospectively identified and studied. Only those who underwent colonoscopy or colon pathologic interpretation were included in the final analysis. RESULTS: From 1994 to 2009, 13 patients met study inclusion criteria. Of the 10 patients who underwent colonoscopy, 9 (90%; 95% confidence interval [CI], 57%-100%) had polyps, and 7 (70%; 95% CI, 39%-90%) were estimated to have more than 50 polyps. Pathologic findings of the colon were reviewed in 11 patients, and the spectrum of tumors included hamartomatous, inflammatory, adenomatous, ganglioneuromatous, hyperplastic, and juvenile polyps. Of the 13 patients, 2 (15%; 95% CI, 3%-43%) had left-sided adenocarcinoma without microsatellite instability. Five (38%) of the 13 patients underwent colectomy secondary to polyp dysplasia. CONCLUSION: Patients with Cowden syndrome have a heavy colon polyp burden with a wide pathologic spectrum, both benign and malignant. The colon polyposis results in a previously unreported morbidity with a high colectomy rate. PMID- 21628614 TI - Palliative medicine consultation for preparedness planning in patients receiving left ventricular assist devices as destination therapy. AB - OBJECTIVE: To assess the benefit of proactive palliative medicine consultation for delineation of goals of care and quality-of-life preferences before implantation of left ventricular assist devices as destination therapy (DT). PATIENTS AND METHODS: We retrospectively reviewed the cases of patients who received DT between January 15, 2009, and January 1, 2010. RESULTS: Of 19 patients identified, 13 (68%) received proactive palliative medicine consultation. Median time of palliative medicine consultation was 1 day before DT implantation (range, 5 days before to 16 days after). Thirteen patients (68%) completed advance directives. The DT implantation team and families reported that preimplantation discussions and goals of care planning made postoperative care more clear and that adverse events were handled more effectively. Currently, palliative medicine involvement in patients receiving DT is viewed as routine by cardiac care specialists. CONCLUSION: Proactive palliative medicine consultation for patients being considered for or being treated with DT improves advance care planning and thus contributes to better overall care of these patients. Our experience highlights focused advance care planning, thorough exploration of goals of care, and expert symptom management and end-of-life care when appropriate. PMID- 21628615 TI - Association between CHADS2risk factors and anticoagulation-related bleeding: a systematic literature review. AB - OBJECTIVE: To determine the strength of evidence supporting an accentuated bleeding risk when patients with CHADS(2) risk factors (chronic heart failure, hypertension, advanced age, diabetes, and prior stroke/transient ischemic attack) receive warfarin. METHODS: A systematic literature search of MEDLINE (January 1, 1950, through December 22, 2009) and Cochrane CENTRAL (through December 22, 2009) was conducted to identify studies that reported multivariate results on the association between CHADS(2) covariates and risk of bleeding in patients receiving warfarin. Each covariate was evaluated for its association with a specific type of bleeding. Individual evaluations were rated as good, fair, or poor using methods consistent with those recommended by the Agency for Healthcare Research and Quality. The strength of the associations between each CHADS(2) covariate and a specific type of bleeding was determined using Grading of Recommendations Assessment, Development and Evaluation criteria as insufficient, very low, low, moderate, or high for the entire body of evidence. RESULTS: Forty one studies were identified, reporting 127 multivariate evaluations of the association between a CHADS(2) covariate and bleeding risk. No CHADS(2) covariate had a high strength of evidence for association with any bleeding type. For the vast majority of evaluations, the strength of evidence between covariates and bleeding was low. Advanced age was the only covariate that had a moderate strength of evidence for association; this was the strongest independent positive predictor for major bleeding. Similar findings were observed regardless of whether all included studies, or only those evaluating patients with atrial fibrillation, were assessed. CONCLUSION: The associations between CHADS(2) covariates and increased bleeding risk were weak, with the exception of age. Given the known association of the CHADS(2) score and stroke risk, the decision to prescribe warfarin should be driven more by patients' risk of stroke than by the risk of bleeding. PMID- 21628616 TI - Mayo Clinic consensus recommendations for the depth of excision in primary cutaneous melanoma. AB - Currently, no data from randomized controlled clinical trials are available to guide the depth of resection for intermediate-thickness primary cutaneous melanoma. Thus, we hypothesized that substantial variability exists in this aspect of surgical care. We have summarized the literature regarding depth of resection and report the results of our survey of surgeons who treat melanoma. Most of the 320 respondents resected down to, but did not include, the muscular fascia (extremity, 71%; trunk, 66%; and head and neck, 62%). However, significant variation exists. We identified variability in our own practice and have elected to standardize this common aspect of routine surgical care across our institution. In light of the lack of evidence to support resection of the deep muscular fascia, we have elected to preserve the muscular fascia as a matter of routine, except when a deep primary melanoma or thin subcutaneous tissue dictates otherwise. PMID- 21628617 TI - Updates on definition, consequences, and management of obstructive sleep apnea. AB - Obstructive sleep apnea (OSA) is a breathing disorder during sleep that has implications beyond disrupted sleep. It is increasingly recognized as an independent risk factor for cardiac, neurologic, and perioperative morbidities. Yet this disorder remains undiagnosed in a substantial portion of our population. It is imperative for all physicians to remain vigilant in identifying patients with signs and symptoms consistent with OSA. This review focuses on updates in the areas of terminology and testing, complications of untreated OSA, perioperative considerations, treatment options, and new developments in this field. PMID- 21628618 TI - Venous thromboembolism in progress. PMID- 21628619 TI - 26-year-old man with recurrent urinary tract infections. PMID- 21628621 TI - More on body fat cutoff points. PMID- 21628620 TI - Antiparasitic therapy. AB - Parasitic diseases affect more than 2 billion people globally and cause substantial morbidity and mortality, particularly among the world's poorest people. This overview focuses on the treatment of the major protozoan and helminth infections in humans. Recent developments in antiparasitic therapy include the expansion of artemisinin-based therapies for malaria, new drugs for soil-transmitted helminths and intestinal protozoa, expansion of the indications for antiparasitic drug treatment in patients with Chagas disease, and the use of combination therapy for leishmaniasis and human African trypanosomiasis. PMID- 21628622 TI - Frequency of herpes zoster recurrence. PMID- 21628624 TI - Differential outcomes between dialysis modalities: purely a reflection of selection bias? PMID- 21628625 TI - A multicentre evaluation of the new Beckman Coulter anti-Mullerian hormone immunoassay (AMH Gen II). AB - BACKGROUND: The measurement of anti-Mullerian hormone (AMH) has been by two commercial enzyme-linked immunosorbent (ELISA) assays: Diagnostics Systems Laboratory (DSL 10-14400) and Immunotech (A11893 IVD EU only). Beckman Coulter has developed a new assay for AMH (AMH Gen II A79765), which uses the DSL antibodies but is standardized to the Immunotech calibration. As a result, comparative data are urgently required between the old DSL assay and its replacement AMH Gen II. METHODS: An evaluation of the AMH Gen II assay was performed at three sites, each with extensive experience of measuring circulating AMH in the adult female. Results were compared with the original DSL ELISA assay. The analysis was performed on a total of 271 patients' samples, approximately 90 at each site. RESULTS: Performance characteristics were evaluated for the AMHGen II assay. Linearity was acceptable with observed values close to the expected (mean recovery 106.3%). The functional sensitivity (20% coefficient of variation), calculated from precision profile data, was 1.5 pmol/L. Within- and between-batch imprecision, assessed over the concentration range of 5-70 pmol/L, were 5.3-11.4% and 3.8-17.3%, respectively. There was good agreement between assays with a Bablok-Passing regression equation AMH Gen II = 1.40 DSL-0.62 pmol/L, r = 0.96, n = 271. CONCLUSIONS: Our results demonstrate that similar precision and excellent between-assay agreement should be obtained when laboratories change from the DSL to the AMH Gen II ELISA and they should expect an increase in AMH values of approximately 40%. PMID- 21628626 TI - Predicting seasonal influenza vaccination among hospital-based nurses. AB - A descriptive cross-sectional online survey of a convenience sample of 202 hospital-based nurses was conducted to explore the factors associated with influenza vaccination. The findings suggest that the independent predictors of influenza vaccination were perception of job as a risk increasing factor (OR = 12.14; 95% CI [1.89, 78.08]), workplace vaccination clinics and campaigns (OR = 2.88; 95% CI [1.12, 7.38]), vaccination in the previous season (OR = 34.80; 95% CI [12.99, 93.28]), viewing vaccination as an inconvenience (OR = 0.22; 95% CI [0.07, 0.67]), and one's belief that the immune system provides better protection than the vaccine (OR = 0.29; 95% CI [0.11, 0.77]). In conclusion, the findings support the existing literature with regards to low vaccination rates among health care providers. Furthermore, the identification of the predictors of influenza vaccination among nurses may assist administrators and policy makers with the implementation of evidence-based vaccination strategies. PMID- 21628627 TI - CAST AWAY, a membrane-associated receptor-like kinase, inhibits organ abscission in Arabidopsis. AB - Receptor-like kinase-mediated cell signaling pathways play fundamental roles in many aspects of plant growth and development. A pair of Arabidopsis (Arabidopsis thaliana) leucine-rich repeat receptor-like kinases (LRR-RLKs), HAESA (HAE) and HAESA-LIKE2 (HSL2), have been shown to activate the cell separation process that leads to organ abscission. Another pair of LRR-RLKs, EVERSHED (EVR) and SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE1, act as inhibitors of abscission, potentially by modulating HAE/HSL2 activity. Cycling of these RLKs to and from the cell surface may be regulated by NEVERSHED (NEV), a membrane trafficking regulator that is essential for organ abscission. We report here the characterization of CAST AWAY (CST), a receptor-like cytoplasmic kinase that acts as a spatial inhibitor of cell separation. Disruption of CST suppresses the abscission defects of nev mutant flowers and restores the discrete identity of the trans-Golgi network in nev abscission zones. After organ shedding, enlarged abscission zones with obscured boundaries are found in nev cst flowers. We show that CST is a dual specificity kinase in vitro and that myristoylation at its amino terminus promotes association with the plasma membrane. Using the bimolecular fluorescence complementation assay, we have detected interactions of CST with HAE and EVR at the plasma membrane of Arabidopsis protoplasts and hypothesize that CST negatively regulates cell separation signaling directly and indirectly. A model integrating the potential roles of receptor-like kinase signaling and membrane trafficking during organ separation is presented. PMID- 21628628 TI - Perturbation of spermine synthase gene expression and transcript profiling provide new insights on the role of the tetraamine spermine in Arabidopsis defense against Pseudomonas viridiflava. AB - The role of the tetraamine spermine in plant defense against pathogens was investigated by using the Arabidopsis (Arabidopsis thaliana)-Pseudomonas viridiflava pathosystem. The effects of perturbations of plant spermine levels on susceptibility to bacterial infection were evaluated in transgenic plants (35S::spermine synthase [SPMS]) that overexpressed the SPMS gene and accumulated spermine, as well as in spms mutants with low spermine levels. The former exhibited higher resistance to P. viridiflava than wild-type plants, while the latter were more susceptible. Exogenous supply of spermine to wild-type plants also increased disease resistance. Increased resistance provided by spermine was partly counteracted by the polyamine oxidase inhibitor SL-11061, demonstrating that the protective effect of spermine partly depends on its oxidation. In addition, global changes in gene expression resulting from perturbations of spermine levels were analyzed by transcript profiling 35S::SPMS-9 and spms-2 plants. Overexpression of 602 genes was detected in 35S::SPMS-9 plants, while 312 genes were down-regulated, as compared to the wild type. In the spms-2 line, 211 and 158 genes were up- and down-regulated, respectively. Analysis of gene ontology term enrichment demonstrated that many genes overexpressed only in 35S::SPMS-9 participate in pathogen perception and defense responses. Notably, several families of disease resistance genes, transcription factors, kinases, and nucleotide- and DNA/RNA-binding proteins were overexpressed in this line. Thus, a number of spermine-responsive genes potentially involved in resistance to P. viridiflava were identified. The obtained results support the idea that spermine contributes to plant resistance to P. viridiflava. PMID- 21628629 TI - The pepper E3 ubiquitin ligase RING1 gene, CaRING1, is required for cell death and the salicylic acid-dependent defense response. AB - Ubiquitination is essential for ubiquitin/proteasome-mediated protein degradation in plant development and defense. Here, we identified a novel E3 ubiquitin ligase RING1 gene, CaRING1, from pepper (Capsicum annuum). In pepper, CaRING1 expression is induced by avirulent Xanthomonas campestris pv vesicatoria infection. CaRING1 contains an amino-terminal transmembrane domain and a carboxyl-terminal RING domain. In addition, it displays in vitro E3 ubiquitin ligase activity, and the RING domain is essential for E3 ubiquitin ligase activity in CaRING1. CaRING1 also localizes to the plasma membrane. In pepper plants, virus-induced gene silencing of CaRING1 confers enhanced susceptibility to avirulent X. campestris pv vesicatoria infection, which is accompanied by compromised hypersensitive cell death, reduced expression of PATHOGENESIS-RELATED1, and lowered salicylic acid levels in leaves. Transient expression of CaRING1 in pepper leaves induces cell death and the defense response that requires the E3 ubiquitin ligase activity of CaRING1. By contrast, overexpression of CaRING1 in Arabidopsis (Arabidopsis thaliana) confers enhanced resistance to hemibiotrophic Pseudomonas syringae pv tomato and biotrophic Hyaloperonospora arabidopsidis infections. Taken together, these results suggest that CaRING1 is involved in the induction of cell death and the regulation of ubiquitination during the defense response to microbial pathogens. PMID- 21628631 TI - Root cortical aerenchyma enhances the growth of maize on soils with suboptimal availability of nitrogen, phosphorus, and potassium. AB - Root cortical aerenchyma (RCA) is induced by hypoxia, drought, and several nutrient deficiencies. Previous research showed that RCA formation reduces the respiration and nutrient content of root tissue. We used SimRoot, a functional structural model, to provide quantitative support for the hypothesis that RCA formation is a useful adaptation to suboptimal availability of phosphorus, nitrogen, and potassium by reducing the metabolic costs of soil exploration in maize (Zea mays). RCA increased the growth of simulated 40-d-old maize plants up to 55%, 54%, or 72% on low nitrogen, phosphorus, or potassium soil, respectively, and reduced critical fertility levels by 13%, 12%, or 7%, respectively. The greater utility of RCA on low-potassium soils is associated with the fact that root growth in potassium-deficient plants was more carbon limited than in phosphorus- and nitrogen-deficient plants. In contrast to potassium-deficient plants, phosphorus- and nitrogen-deficient plants allocate more carbon to the root system as the deficiency develops. The utility of RCA also depended on other root phenes and environmental factors. On low-phosphorus soils (7.5 MUM), the utility of RCA was 2.9 times greater in plants with increased lateral branching density than in plants with normal branching. On low-nitrate soils, the utility of RCA formation was 56% greater in coarser soils with high nitrate leaching. Large genetic variation in RCA formation and the utility of RCA for a range of stresses position RCA as an interesting crop-breeding target for enhanced soil resource acquisition. PMID- 21628630 TI - Arabidopsis Pht1;5 mobilizes phosphate between source and sink organs and influences the interaction between phosphate homeostasis and ethylene signaling. AB - Phosphorus (P) remobilization in plants is required for continuous growth and development. The Arabidopsis (Arabidopsis thaliana) inorganic phosphate (Pi) transporter Pht1;5 has been implicated in mobilizing stored Pi out of older leaves. In this study, we used a reverse genetics approach to study the role of Pht1;5 in Pi homeostasis. Under low-Pi conditions, Pht1;5 loss of function (pht1;5-1) resulted in reduced P allocation to shoots and elevated transcript levels for several Pi starvation-response genes. Under Pi-replete conditions, pht1;5-1 had higher shoot P content compared with the wild type but had reduced P content in roots. Constitutive overexpression of Pht1;5 had the opposite effect on P distribution: namely, lower P levels in shoots compared with the wild type but higher P content in roots. Pht1;5 overexpression also resulted in altered Pi remobilization, as evidenced by a greater than 2-fold increase in the accumulation of Pi in siliques, premature senescence, and an increase in transcript levels of genes involved in Pi scavenging. Furthermore, Pht1;5 overexpressors exhibited increased root hair formation and reduced primary root growth that could be rescued by the application of silver nitrate (ethylene perception inhibitor) or aminoethoxyvinylglycine (ethylene biosynthesis inhibitor), respectively. Together, these data indicate that Pht1;5 plays a critical role in mobilizing Pi from P source to sink organs in accordance with developmental cues and P status. The study also provides evidence for a link between Pi and ethylene signaling pathways. PMID- 21628632 TI - Body mass index and long-term mortality in an elderly Mediterranean population. AB - OBJECTIVES: To describe the relationship of body mass index and mortality in older adults, examining the influence of sex and cardiovascular morbidity. METHODS: Sixteen-year cohort of a population sample of 1,008 people aged 65 and over. BMI mortality hazard ratios are estimated controlling for age, sex, education, physical activity, smoking, chronic conditions, and ADL (activities of daily living) disability. RESULTS: At baseline the median BMI is 26.8 (Interquartile range: 24.2-29.7 Kg/m(2)). Findings show that during 16 years there were 672 deaths. The U-shaped curve of the mortality hazard by BMI is wide. The minimum mortality occur at BMI = 30.5 Kg/m(2). Findings show that men had lower mortality risk with increasing BMI and that cardiovascular disease was associated with high mortality in the low-BMI category. DISCUSSION: Underweight is a risk factor for mortality among elderly people, whereas overweight and mild obesity are associated with the lowest mortality particularly among men and those with cardiovascular morbidity. PMID- 21628633 TI - Circulating uncarboxylated matrix gla protein is associated with vitamin K nutritional status, but not coronary artery calcium, in older adults. AB - Matrix Gla protein (MGP) is a calcification inhibitor in vascular tissue that must be carboxylated by vitamin K to function. Evidence suggests circulating uncarboxylated MGP (ucMGP) is elevated in persons with disease characterized by vascular calcification. The primary purpose of this study was to determine cross sectional and longitudinal associations between plasma ucMGP, vitamin K status, and coronary artery calcium (CAC) in older adults without coronary heart disease. Genetic determinants of ucMGP were also explored. Cross-sectional associations among baseline plasma ucMGP, vitamin K status biomarkers [plasma phylloquinone, uncarboxylated prothrombin (PIVKA-II), serum uncarboxylated osteocalcin (%ucOC)], CAC, and plausible genetic polymorphisms were examined in 438 community-dwelling adults (60-80 y, 59% women). The effect of phylloquinone supplementation (500 MUg/d) for 3 y on plasma ucMGP was determined among 374 participants. At baseline, plasma phylloquinone was lower and %ucOC and PIVKA-II were greater across higher plasma ucMGP quartiles (all P < 0.001, age-adjusted). Major allele homozygotes for MGP rs1800801 and rs4236 had higher plasma ucMGP than heterozygotes or minor allele homozygotes. (P <= 0.004). The decrease in plasma ucMGP was greater in the 190 participants who received phylloquinone (mean +/- SD) (-345 +/- 251 pmol/L) than in the 184 who did not (-40 +/- 196 pmol/L) (P < 0.0001). CAC did not differ according to ucMGP quartile (P = 0.35, age-adjusted). In the phylloquinone-supplemented group, the 3-y change in ucMGP was not associated with the 3-y change in CAC [unstandard beta (SE) = -0.02 (0.02); P = 0.44]. Plasma ucMGP was associated with vitamin K status biomarkers and was reduced following phylloquinone supplementation, suggesting it may be a useful marker of vitamin K status in vascular tissue. Plasma ucMGP did not reflect CAC in healthy older adults. PMID- 21628634 TI - CLA-enriched diet containing t10,c12-CLA alters bile acid homeostasis and increases the risk of cholelithiasis in mice. AB - Mice fed a mixture of CLA containing t10,c12-CLA lose fat mass and develop hyperinsulinemia and hepatic steatosis due to an accumulation of TG and cholesterol. Because cholesterol is the precursor in bile acid (BA) synthesis, we investigated whether t10,c12-CLA alters BA metabolism. In Expt. 1, female C57Bl/6J mice were fed a standard diet for 28 d supplemented with a CLA mixture (1 g/100 g) or not (controls). In Expt. 2, the feeding period was reduced to 4, 6, and 10 d. In Expt. 3, mice were fed a diet supplemented with linoleic acid, c9,t11-CLA, or t10,c12-CLA (0.4 g/100 g) for 28 d. In Expt. 1, the BA pool size was greater in CLA-fed mice than in controls and the entero-hepatic circulation of BA was altered due to greater BA synthesis and ileal reclamation. This resulted from higher hepatic cholesterol 7alpha-hydroxylase (CYP7A1) and ileal apical sodium BA transporter expressions in CLA-fed mice. Furthermore, hepatic Na(+)/taurocholate co-transporting polypeptide (NTCP) (-52%) and bile salt export pump (BSEP) (-77%) protein levels were lower in CLA-fed mice than in controls, leading to a greater accumulation of BA in the plasma (+500%); also, the cholesterol saturation index and the concentration of hydrophobic BA in the bile were greater in CLA-fed mice, changes associated with the presence of cholesterol crystals. Expt. 2 suggests that CLA-mediated changes were caused by hyperinsulinemia, which occurred after 6 d of the CLA diet before NTCP and BSEP mRNA downregulation (10 d). Expt. 3 demonstrated that only t10,c12-CLA altered NTCP and BSEP mRNA levels. In conclusion, t10,c12-CLA alters BA homeostasis and increases the risk of cholelithiasis in mice. PMID- 21628635 TI - Starch with high amylose content and low in vitro digestibility increases intestinal nutrient flow and microbial fermentation and selectively promotes bifidobacteria in pigs. AB - Diets containing different starch types can affect enzymatic digestion of starch and thereby starch availability for microbial fermentation in the gut. However, the role of starch chemistry in nutrient digestion and flow and microbial profile has been poorly explained. Eight ileal-cannulated pigs (29.4 +/- 0.9 kg body weight) were fed 4 diets containing 70% purified starch (amylose content, <5, 20, 28, and 63%; reflected by in vitro maximal digestion rate; 1.06, 0.73, 0.38, and 0.22%/min, respectively) in a replicated 4 * 4 Latin square. Ileal and fecal starch output, postileal crude protein yield, fecal total SCFA and total butyrate content, and gene copies of Bifidobacterium spp. in feces were higher (P < 0.05) when pigs consumed the slowly digestible starch diet than the remaining 3 starch diets. The in vitro starch digestion rate had a negative, nonlinear relationship with ileal starch flow (R(2) = 0.98; P < 0.001). Ileal starch flow was positively related to Bifidobacterium spp. (R(2) = 0.27; P < 0.01), Lactobacillus group (R(2) = 0.22; P < 0.01), and total butyrate content (R(2) = 0.46; P < 0.01) but was not related to Enterobacteriaceae (R(2) < 0.00; P = 0.92). In conclusion, starch with high amylose content and low in vitro digestibility increased postileal nutrient flow and microbial fermentation and selectively promoted Bifidobacterium spp. in the distal gut. PMID- 21628636 TI - A priori-defined dietary patterns are associated with reduced risk of stroke in a large Italian cohort. AB - Stroke is a major cause of death. Several foods and nutrients have been linked to stroke, but their effects may be best investigated considering the entire diet. In the present EPICOR study, we investigated the association between stroke and adherence to 4 a priori-defined dietary patterns: Healthy Eating Index 2005 (HEI 2005), Dietary Approaches to Stop Hypertension (DASH), Greek Mediterranean Index, and Italian Mediterranean Index. We followed 40,681 volunteers and estimated the HR and 95%CI for stroke according to dietary pattern by using multivariate Cox models with adjustment for risk factors. During a mean follow-up of 7.9 y, 178 stroke cases were diagnosed (100 ischemic, 47 hemorrhagic). Scores of 3 dietary patterns (not HEI) were inversely associated with risk of all types of stroke, with the strongest association for the Italian Index [HR = 0.47 (95%CI = 0.30 0.75); third vs. first tertile]. All patterns were significantly inversely associated with ischemic stroke except the Greek Index, with the strongest association for the Italian Index [HR = 0.37 (95%CI = 0.19-0.70); third vs. first tertile]. Only the Italian Index tended to be inversely associated with hemorrhagic stroke [HR = 0.51 (95%CI = 0.22-1.20); P = 0.07)]. These epidemiological findings suggest that adherence to any one dietary pattern investigated would protect against at least one type of stroke. For our Italian population, a diet with a high score on the Italian Index was associated with the greatest risk reduction, probably because it was conceived to capture healthy eating in the context of foods typically available in Italy. PMID- 21628637 TI - Surgical trends in Bankart repair: an analysis of data from the American Board of Orthopaedic Surgery certification examination. AB - BACKGROUND: Arthroscopic Bankart repair emerged in the 1990s as a minimally invasive alternative to open repair. The optimal technique of surgical stabilization of the unstable glenohumeral joint remains controversial. HYPOTHESIS: A review of the American Board of Orthopaedic Surgery (ABOS) data would show a trend toward an increasing number of arthroscopic versus open Bankart procedures. STUDY DESIGN: Descriptive epidemiology study. METHODS: A query of the ABOS database for all cases of open or arthroscopic Bankart repair from 2003 through 2008 was performed, as the CPT (Current Procedural Terminology) codes for arthroscopic repair were introduced in 2003. All cases coded with CPT codes for arthroscopic Bankart repair (29806) or open Bankart repair (23455) were reviewed. Additional data were obtained on the surgeons (year of procedure, geographic location, fellowship training, subspecialty examination area) as well as the patients (age, gender, follow-up length, complications, objective outcome measures [pain, deformity, function, and satisfaction]). RESULTS: From 2003 to 2008, a total of 4562 Bankart repair cases were reported, composing 8.6% of the total number of shoulder surgery cases in the ABOS database. From 2003 to 2005, 71.2% of Bankart repairs were arthroscopic, compared with 87.7% between 2006 and 2008 (P < .0001). Surgeons having obtained subspecialty training in sports medicine performed the majority (65.3%) of Bankart repairs. Over the entire period, sports-trained surgeons also performed a higher proportion of arthroscopic repairs (84.1%) compared with surgeons without this training (71.9%) (P < .0001). However, by 2008 both non-fellowship-trained and sports medicine fellowship-trained surgeons performed arthroscopic repair in 90% of cases. Surgeons in the Northeast region performed a significantly greater proportion of arthroscopic Bankart repairs (84.7%) than did surgeons in other regions (78.6%) (P < .0001) from 2003 to 2008. The most commonly reported complications were nerve palsy/injury and dislocation, with a rate of nerve injury of 2.2% in the open group compared to 0.3% in the arthroscopic group (P < .0001), and dislocation rate of 1.2% with open stabilization compared with 0.4% arthroscopically (P = .0039). CONCLUSION: Review of the ABOS data shows a trend toward arthroscopic shoulder stabilization over time, with the use of open repair declining. Reported complications were lower overall in the arthroscopic stabilization group when compared with open surgeries. PMID- 21628638 TI - Characterization of subchondral bone repair for marrow-stimulated chondral defects and its relationship to articular cartilage resurfacing. AB - BACKGROUND: Microfracture and drilling are bone marrow-stimulation techniques that initiate cartilage repair by providing access to cell populations in subchondral bone marrow. This study examined the effect of hole depth and of microfracture versus drilling on subchondral bone repair and cartilage repair in full-thickness chondral defects. HYPOTHESES: Repaired subchondral bone does not reconstitute its native structure and exhibits atypical morphologic features. Drilling deeper induces greater bone remodeling and is related to improved cartilage repair. STUDY DESIGN: Controlled laboratory study. METHODS: Trochlear cartilage defects debrided of the calcified layer were prepared bilaterally in 16 skeletally mature rabbits. Drill holes were made to a depth of 2 mm or 6 mm and microfracture holes to 2 mm. Animals were sacrificed 3 months postoperatively, and joints were scanned by micro-computed tomography before histoprocessing. Bone repair was assessed with a novel scoring system and by 3-dimentional micro computed tomography and compared with intact controls. Correlation of subchondral bone features to cartilage repair outcome was performed. RESULTS: Although surgical holes were partly repaired with mineralized tissue, atypical features such as residual holes, cysts, and bony overgrowth were frequently observed. For all treatment groups, repair led to an average bone volume density similar to that of the controls but the repair bone was more porous and branched as shown by significantly higher bone surface area density and connectivity density. Deeper versus shallower drilling induced a larger region of repairing and remodeling subchondral bone that positively correlated with improved cartilage repair. CONCLUSION: Incomplete reconstitution of normal bone structure and continued remodeling occurred in chondral defects 3 months after bone marrow stimulation. Deep drilling induced a larger volume of repairing and remodeling bone, which appeared beneficial for chondral repair. CLINICAL RELEVANCE: Bone marrow stimulation does not reconstitute normal bone structure. Strategies that increase subchondral bone involvement in marrow stimulation could further benefit cartilage repair. PMID- 21628639 TI - Quantitative prediction of human pregnane X receptor and cytochrome P450 3A4 mediated drug-drug interaction in a novel multiple humanized mouse line. AB - Cytochrome P450 (P450) 3A4 is the predominant P450 enzyme expressed in human liver and intestine, and it is involved in the metabolism of approximately 50% of clinically used drugs. Because of the differences in the multiplicity of CYP3A genes and the poor correlation of substrate specificity of CYP3A proteins between species, the extrapolation of CYP3A-mediated metabolism of a drug from animals to man is difficult. This situation is further complicated by the fact that the predictability of the clinically common drug-drug interaction of pregnane X receptor (PXR)-mediated CYP3A4 induction by animal studies is limited as a result of marked species differences in the interaction of many drugs with this receptor. Here we describe a novel multiple humanized mouse line that combines a humanization for PXR, the closely related constitutive androstane receptor, and a replacement of the mouse Cyp3a cluster with a large human genomic region carrying CYP3A4 and CYP3A7. We provide evidence that this model shows a human-like CYP3A4 induction response to different PXR activators, that it allows the ranking of these activators according to their potency to induce CYP3A4 expression in the human liver, and that it provides an experimental approach to quantitatively predict PXR/CYP3A4-mediated drug-drug interactions in humans. PMID- 21628640 TI - The reduction in fatigue crack growth resistance of dentin with depth. AB - The fatigue crack growth resistance of dentin was characterized as a function of depth from the dentino-enamel junction. Compact tension (CT) specimens were prepared from the crowns of third molars in the deep, middle, and peripheral dentin. The microstructure was quantified in terms of the average tubule dimensions and density. Fatigue cracks were grown in-plane with the tubules and characterized in terms of the initiation and growth responses. Deep dentin exhibited the lowest resistance to the initiation of fatigue crack growth, as indicated by the stress intensity threshold (DeltaK(th) ~ 0.8 MPa*m(0.5)) and the highest incremental fatigue crack growth rate (over 1000 times that in peripheral dentin). Cracks in deep dentin underwent incremental extension under cyclic stresses that were 40% lower than those required in peripheral dentin. The average fatigue crack growth rates increased significantly with tubule density, indicating the importance of microstructure on the potential for tooth fracture. Molars with deep restorations are more likely to suffer from the cracked-tooth syndrome, because of the lower fatigue crack growth resistance of deep dentin. PMID- 21628641 TI - Parotid secretory protein binds phosphatidylinositol (3,4) bisphosphate. AB - Molecular interactions that direct trafficking of secreted proteins are not well described in salivary glands. Here, we report that the soluble cargo protein Parotid Secretory Protein (PSP) is bound to the membranes of secretory granules isolated from rat parotids. This is apparently due to specific interaction with phosphatidylinositol phosphates (PtdInsP). PSP binds PtdIns(3,4)P(2), 10-fold greater than PtdIns(3,5)P(2) or PtdIns(4)P, and does not bind PtdIns(3)P or PtdIns(5)P. Human PSP synthesized in vitro also binds PtdIns(3,4)P(2). Bacterially expressed rat PSP binds PtdIns(3,4)P(2) with a K(d) of 2.4 x 10(-11) M. Other major secretory proteins (amylase, proline-rich protein) are not bound to isolated granule membranes and do not bind phosphatidylinositol phosphates. Immunofluorescence shows PtdIns(3,4)P(2) at the secretory granules, and fluorescent PtdIns(3,4)P(2) can flip from the outer leaflet to the inner leaflet of the membrane. Binding of PSP to PtdInsPs may contribute to sorting during the formation of the secretory granules, or sorting by retention during maturation of the granules. PMID- 21628642 TI - Metaphyseal fixation in revision total knee arthroplasty: indications and techniques. AB - The need for revision total knee arthroplasty (TKA) is on the rise. Challenges to attaining durable, stable, well-functioning revision TKA include bony deficiency, periarticular osteopenia, deformity, and soft-tissue imbalance. Defect management often requires the use of stems, cement, metal augmentation, or allograft. Recently, there has been interest in obtaining fixation in the metaphyseal region in an attempt to improve construct stability while managing bony deficiency. Often, the metaphyseal bone is well vascularized, which provides an opportunity for additional fixation with cement, allograft, trabecular metal cones, or stepped porous-coated sleeves. Multiple series have documented good survivorship at short-term follow-up with trabecular metal cones and porous-coated sleeves. These newer technologies offer biologic fixation and are useful for treating bony defects that are not easily managed with other methods. Long-term studies are needed to determine the durability of these constructs. Concerns persist regarding stress shielding and difficulty of removal. Familiarity with the rationale and strategies for metaphyseal fixation in revision TKA is a valuable addition to the armamentarium of the revision surgeon. PMID- 21628643 TI - Cervical spine trauma in children and adults: perioperative considerations. AB - A wide spectrum of cervical spine injuries, including stable and unstable injuries with and without neurologic compromise, account for a large percentage of emergency department visits. Effective treatment of the polytrauma patient with cervical spine injury requires knowledge of cervical spine anatomy and the pathophysiology of spinal cord injury, as well as techniques for cervical spine stabilization, intraoperative positioning, and airway management. The orthopaedic surgeon must oversee patient care and coordinate treatment with emergency department physicians and anesthesia services in both the acute and subacute settings. Children are particularly susceptible to substantial destabilizing cervical injuries and must be treated with a high degree of caution. The surgeon must understand the unique anatomic and biomechanical properties associated with the pediatric cervical spine as well as injury patterns and stabilization techniques specific to this patient population. PMID- 21628644 TI - Complications of total elbow arthroplasty. AB - Modifications in implant design and improvements in surgical technique have expanded the applications of total elbow arthroplasty. Complications associated with reconstructive elbow surgery persist, however, often leading to profound and sometimes nonsalvageable disability. The most recognized complications include implant loosening, periprosthetic fracture, implant failure, infection, triceps insufficiency, and nerve palsy. Although far fewer elbow arthroplasties than lower extremity arthroplasties are performed, the proportion of complications is greater with elbow arthroplasty, and the outcomes of secondary reconstruction are less favorable. PMID- 21628645 TI - Hip instability. AB - Understanding of the etiology and pathology of hip instability has increased in recent years as new information has emerged regarding the disease processes of the hip. Hip instability, heretofore considered uncommon in clinical practice, is increasingly recognized as a pathologic entity. Instability may be classified as traumatic or atraumatic, and diagnosis is made based on patient history, physical examination, and imaging studies. Plain radiography, MRI, MRI arthrography, and hip instability tests (eg, posterior impingement, dial) can be used to confirm the presence of instability. Nonsurgical management options include physical therapy and protected weight bearing. Surgical intervention, whether arthroscopic or open, is required for large acetabular fractures and refractory instability. Knowledge of the etiology and evolving research of hip instability is essential to understand the spectrum of hip disease. PMID- 21628646 TI - Management of lateral humeral condylar fracture in children. AB - Lateral condylar fractures constitute 12% to 20% of all pediatric distal humerus fractures. These fractures are easily missed and when not managed appropriately can displace. Missed fracture is a common cause of nonunion and deformity; thus, a high index of suspicion and adequate clinical and radiographic evaluation are required. Displaced fractures are associated with a high rate of nonunion. Nondisplaced fractures or those displaced <=2 mm are managed with cast immobilization and frequent radiographic follow-up. Fractures displaced >2 mm are managed with surgical fixation. Successful outcomes have been reported with closed reduction, open reduction, and arthroscopically assisted techniques. Complications associated with pediatric lateral condylar fracture include cubitus varus, cubitus valgus, fishtail deformity, and tardy ulnar nerve palsy. PMID- 21628647 TI - Four common types of bursitis: diagnosis and management. AB - Bursitis is a common cause of musculoskeletal pain and often prompts orthopaedic consultation. Bursitis must be distinguished from arthritis, fracture, tendinitis, and nerve pathology. Common types of bursitis include prepatellar, olecranon, trochanteric, and retrocalcaneal. Most patients respond to nonsurgical management, including ice, activity modification, and nonsteroidal anti inflammatory drugs. In cases of septic bursitis, oral antibiotics may be administered. Local corticosteroid injection may be used in the management of prepatellar and olecranon bursitis; however, steroid injection into the retrocalcaneal bursa may adversely affect the biomechanical properties of the Achilles tendon. Surgical intervention may be required for recalcitrant bursitis, such as refractory trochanteric bursitis. PMID- 21628648 TI - Optimizing the management of rotator cuff problems. AB - Of the 31 recommendations made by the work group, 19 were determined to be inconclusive because of the absence of definitive evidence. Of the remaining recommendations, four were classified as moderate grade, six as weak, and two as consensus statements of expert opinion. The four moderate-grade recommendations include suggestions that exercise and nonsteroidal anti-inflammatory drugs be used to manage rotator cuff symptoms in the absence of a full-thickness tear, that routine acromioplasty is not required at the time of rotator cuff repair, that non-cross-linked, porcine small intestine submucosal xenograft patches not be used to manage rotator cuff tears, and that surgeons can advise patients that workers' compensation status correlates with less favorable outcomes after rotator cuff surgery. PMID- 21628649 TI - AAOS clinical practice guideline: optimizing the management of rotator cuff problems. PMID- 21628650 TI - Adjusting for heterogeneity of malaria transmission in longitudinal studies. PMID- 21628651 TI - Whipple disease research accelerates. PMID- 21628652 TI - Francisella tularensis: a red-blooded pathogen. PMID- 21628653 TI - Effect of the pre-erythrocytic candidate malaria vaccine RTS,S/AS01E on blood stage immunity in young children. AB - BACKGROUND: RTS,S/AS01(E) is the lead candidate malaria vaccine and confers pre erythrocytic immunity. Vaccination may therefore impact acquired immunity to blood-stage malaria parasites after natural infection. METHODS: We measured, by enzyme-linked immunosorbent assay, antibodies to 4 Plasmodium falciparum merozoite antigens (AMA-1, MSP-1(42), EBA-175, and MSP-3) and by growth inhibitory activity (GIA) using 2 parasite clones (FV0 and 3D7) at 4 times on 860 children who were randomized to receive with RTS,S/AS01(E) or a control vaccine. RESULTS: Antibody concentrations to AMA-1, EBA-175, and MSP-1(42) decreased with age during the first year of life, then increased to 32 months of age. Anti-MSP-3 antibody concentrations gradually increased, and GIA gradually decreased up to 32 months. Vaccination with RTS,S/AS01(E) resulted in modest reductions in AMA-1, EBA-175, MSP-1(42), and MSP-3 antibody concentrations and no significant change in GIA. Increasing anti-merozoite antibody concentrations and GIA were prospectively associated with increased risk of clinical malaria. CONCLUSIONS: Vaccination with RTS,S/AS01E reduces exposure to blood-stage parasites and, thus, reduces anti-merozoite antigen antibody concentrations. However, in this study, these antibodies were not correlates of clinical immunity to malaria. Instead, heterogeneous exposure led to confounded, positive associations between increasing antibody concentration and increasing risk of clinical malaria. PMID- 21628654 TI - Antibodies to Plasmodium falciparum antigens predict a higher risk of malaria but protection from symptoms once parasitemic. AB - BACKGROUND: Associations between antibody responses to Plasmodium falciparum antigens and protection against symptomatic malaria have been difficult to ascertain, in part because antibodies are potential markers of both exposure to P. falciparum and protection against disease. METHODS: We measured IgG responses to P. falciparum circumsporozoite protein, liver-stage antigen 1, apical-membrane antigen 1 (AMA-1), and merozoite surface proteins (MSP) 1 and 3, in children in Kampala, Uganda, and measured incidence of malaria before and after antibody measurement. RESULTS: Stronger responses to all 5 antigens were associated with an increased risk of clinical malaria (P < .01) because of confounding with prior exposure to P. falciparum. However, with use of another assessment, risk of clinical malaria once parasitemic, stronger responses to AMA-1, MSP-1, and MSP-3 were associated with protection (odds ratios, 0.34, 0.36, and 0.31, respectively, per 10-fold increase; P < .01). Analyses assessing antibodies in combination suggested that any protective effect of antibodies was overestimated by associations between individual responses and protection. CONCLUSIONS: Using the risk of symptomatic malaria once parasitemic as an outcome may improve detection of associations between immune responses and protection from disease. Immunoepidemiology studies designed to detect mechanisms of immune protection should integrate prior exposure into the analysis and evaluate multiple immune responses. PMID- 21628655 TI - Transcription of var genes other than var2csa in Plasmodium falciparum parasites infecting Mozambican pregnant women. AB - BACKGROUND: Increased susceptibility to Plasmodium falciparum infection during pregnancy has been attributed to the accumulation of infected erythrocytes in the placenta. This phenomenon is mediated by a var gene coding for VAR2CSA, which adheres to chondroitin sulphate A. However, the contribution of parasites transcribing other var genes to maternal infections has not been well characterized. METHODS: Transcription of var2csa and var groups A, B, and C was measured by real-time polymerase chain reaction in 30 placental and 21 peripheral P. falciparum isolates from pregnant women and in 42 isolates from nonpregnant adults and children. Associations of infections with non-var2csa isolates with maternal parasitemia and immune responses were assessed. RESULTS: Placental parasites showed the highest levels of var2csa. ABC var genes were transcribed by 20 (67%) of 30 placental isolates and were associated with higher parasitemia compared with infections by parasites only transcribing var2csa (P = .004). Peripheral isolates from pregnant women transcribed ABC var genes at levels similar to those of parasites infecting nonpregnant adults with clinical malaria (P[varA] = .420, P[varB] = .808, and P[varC] = .619). CONCLUSIONS: Transcripts of var2csa are abundant in pregnancy-associated P. falciparum infections; however, ABC var types are also common, especially in peripheral blood, with transcription levels similar to those of infections out of pregnancy. These findings are of interest for the design of malaria vaccines for pregnant women. PMID- 21628656 TI - Endogenous IL-13 plays a crucial role in liver granuloma maturation during Leishmania donovani infection, independent of IL-4Ralpha-responsive macrophages and neutrophils. AB - Previous studies comparing interleukin 4 receptor alpha (IL-4Ralpha)(-/-) and interleukin 4 (IL-4)(-/-) BALB/c mice have indicated that interleukin 13 (IL-13), whose receptor shares the IL-4Ralpha subunit with IL-4, plays a protective role during visceral leishmaniasis. We demonstrate that IL-13(-/-) BALB/c mice were less able to control hepatic growth of Leishmania donovani compared with wild type mice. This correlated with significantly retarded granuloma maturation in IL 13(-/-) mice, defective interferon gamma (IFN-gamma) production, and elevated IL 4 and interleukin 10 (IL-10) levels. L. donovani-infected IL-13(-/-) mice also responded poorly to sodium stibogluconate-mediated chemotherapy compared with wild-type BALB/c mice. Because murine lymphocytes do not have IL-13 receptors, we examined the ability of macrophage/neutrophil-specific IL-4Ralpha(-/-) mice to control primary infection with L. donovani and to respond to chemotherapy. Macrophage/neutrophil-specific IL-4Ralpha(-/-) mice were as resistant to leishmaniasis as wild-type mice, and chemotherapy retained its efficacy. Consequently, in L. donovani infected BALB/c mice, IL-13 promotes hepatic granuloma formation and controls parasite burdens independently of direct effects on macrophages/neutrophils. PMID- 21628657 TI - An experimental mouse model to establish Tropheryma whipplei as a diarrheal agent. AB - Tropheryma whipplei has long been considered as a rare bacterium causing a rare disease, Whipple's disease. However, recent advances now suggest that T. whipplei is a ubiquitous environmental bacterium that may cause gastroenteritis, commonly associated with viral pathogens. We developed an animal model to support this hypothesis. We found that orally given T. whipplei induced diarrhea in mice, without spreading into the intestines. Aggravating factors, such as damage to the intestinal mucosa, favored bacterial spreading. Indeed, bacterial presence was prolonged in stools of dextran sulfate-treated mice, and bacteria were detected in the colon. This resulted in an immune response, with T. whipplei-specific serum IgM and IgG and fecal IgA, as measured by newly introduced immuno polymerase chain reaction technique. Our results confirm that T. whipplei is an agent causing gastroenteritis and suggest that existing mucosal damage may favor bacterial invasion of tissues. PMID- 21628658 TI - Invasion of erythrocytes by Francisella tularensis. AB - Francisella tularensis is the causative agent of tularemia and is classified as a category A biodefense agent by the Centers for Disease Control and Prevention because of its highly infectious nature. F. tularensis infects leukocytes and exhibits an extracellular phase in the blood of the host. It is unknown, however, whether F. tularensis can infect erythrocytes; thus, we examined this possibility in vivo and in vitro. In the murine model of pulmonary type A tularemia, we showed the presence of intraerythrocytic bacteria by double-immunofluorescence microscopy and ex vivo gentamicin protection of the purified erythrocyte fraction. In vitro, F. tularensis invaded human erythrocytes, as shown in the gentamicin protection assays, double-immunofluorescence microscopy, flow cytometry, scanning electron microscopy, and transmission electron microscopy with immunogold labeling of the bacteria. Additional in vitro tests indicated that serum complement-dependent and complement-independent mechanisms contribute to erythrocyte invasion. Our results reveal a novel intraerythrocytic phase during F. tularensis infection. PMID- 21628659 TI - Volunteer challenge with enterotoxigenic Escherichia coli that express intestinal colonization factor fimbriae CS17 and CS19. AB - Human challenges with enterotoxigenic Escherichia coli (ETEC) have broadened our understanding of this important enteropathogen. We report findings from the first challenge studies using ETEC-expressing colonization factor fimbria CS17 and CS19. LSN03-016011/A (LT, CS17) elicited a dose-dependent effect, with the upper dose (6 * 10(9) organisms) causing diarrhea in 88% of recipients. WS0115A (LTSTp, CS19) also showed a dose response, with a 44% diarrhea rate at 9 * 10(9) organisms. Both strains elicited homologous antifimbrial and anti-LT antibody seroconversion. These studies establish the relative pathogenicity of ETEC expressing newer class 5 fimbriae and suggest suitability of the LT|CS17-ETEC challenge model for interventional trials. PMID- 21628660 TI - Decorin binding by DbpA and B of Borrelia garinii, Borrelia afzelii, and Borrelia burgdorferi sensu Stricto. AB - BACKGROUND: Decorin adherence is crucial in the pathogenesis of Lyme borreliosis. Decorin-binding proteins (Dbp) A and B are the adhesins that mediate this interaction. DbpA and B of Borrelia garinii, Borrelia afzelii, and Borrelia burgdorferi sensu stricto (ss) differ in their amino acid sequence, but little attention has been paid to the potential difference in their decorin binding. METHODS: We expressed recombinant DbpA and DbpB of B. garinii, B. afzelii, and B. burgdorferi ss and studied their binding to decorin. We also generated recombinant Borrelia strains to study the role of DbpA and DbpB in the adhesion of live spirochetes to decorin and decorin-expressing cells. RESULTS. Recombinant DbpA of B. garinii and DbpB of B. garinii and B. burgdorferi ss showed strong binding to decorin, whereas DbpA of B. burgdorferi ss and both DbpA and DbpB of B. afzelii exhibited no or only minor binding activity. DbpA and DbpB of B. garinii and B. burgdorferi ss also supported the adhesion of whole spirochetes to decorin and decorin-expressing cells, whereas DbpA and DbpB of B. afzelii did not exhibit this activity. CONCLUSIONS: Dbp A and B of B. garinii and B. burgdorferi ss mediate the interaction between the spirochete and decorin, whereas the same adhesins of B. afzelii show only negligible activity. PMID- 21628661 TI - A systematic review and meta-analysis of interventions to prevent hepatitis C virus infection in people who inject drugs. AB - INTRODUCTION: High rates of hepatitis C virus (HCV) transmission are found in samples of people who inject drugs (PWID) throughout the world. The objective of this paper was to meta-analyze the effects of risk-reduction interventions on HCV seroconversion and identify the most effective intervention types. METHODS: We performed a systematic review and meta-analysis of published and unpublished studies. Eligible studies reported on the association between participation in interventions intended to reduce unsafe drug injection and HCV seroconversion in samples of PWID. RESULTS: The meta-analysis included 26 eligible studies of behavioral interventions, substance-use treatment, syringe access, syringe disinfection, and multicomponent interventions. Interventions using multiple combined strategies reduced risk of seroconversion by 75% (pooled relative risk, .25; 95% confidence interval, .07-.83). Effects of single-method interventions ranged from .6 to 1.6. CONCLUSIONS: Interventions using strategies that combined substance-use treatment and support for safe injection were most effective at reducing HCV seroconversion. Determining the effective dose and combination of interventions for specific subgroups of PWID is a research priority. However, our meta-analysis shows that HCV infection can be prevented in PWID. PMID- 21628662 TI - IL28B but not ITPA polymorphism is predictive of response to pegylated interferon, ribavirin, and telaprevir triple therapy in patients with genotype 1 hepatitis C. AB - BACKGROUND: Pegylated interferon, ribavirin, and telaprevir triple therapy is a new strategy expected to eradicate the hepatitis C virus (HCV) even in patients infected with difficult-to-treat genotype 1 strains, although adverse effects, such as anemia and rash, are frequent. METHODS: We assessed efficacy and predictive factors for sustained virological response (SVR) for triple therapy in 94 Japanese patients with HCV genotype 1. We included recently identified predictive factors, such as IL28B and ITPA polymorphism, and substitutions in the HCV core and NS5A proteins. RESULTS: Patients treated with triple therapy achieved comparatively high SVR rates (73%), especially among treatment-naive patients (80%). Of note, however, patients who experienced relapse during prior pegylated interferon plus ribavirin combination therapy were highly likely to achieve SVR while receiving triple therapy (93%); conversely, prior nonresponders were much less likely to respond to triple therapy (32%). In addition to prior treatment response, IL28B SNP genotype and rapid viral response were significant independent predictors for SVR. Patients with the anemia-susceptible ITPA SNP rs1127354 genotype typically required ribavirin dose reduction earlier than did patients with other genotypes. CONCLUSIONS: Analysis of predictive factors identified IL28B SNP, rapid viral response, and transient response to previous therapy as significant independent predictors of SVR after triple therapy. PMID- 21628663 TI - A competitive serological assay shows naturally acquired immunity to human papillomavirus infections in the Guanacaste Natural History Study. AB - BACKGROUND: A competitive Luminex Immunoassay (cLIA) has been developed to measure neutralizing antibodies against human papillomavirus (HPV) types 6, 11, 16 and 18. METHODS: In a cohort of 974 women from the Guanacaste Natural History Study, we studied the relationship of baseline cLIA and virus-like particle (VLP) enzyme-linked immunosorbent assay (ELISA) (HPV16 and HPV18 only) seropositivity to measures of HPV exposure, HPV DNA positivity, number of sexual partners, cytology findings, and age. We then studied immunity against subsequent infection with HPV6, 11, 16, 18 and related types over a 7-year period. RESULTS: cLIA seroprevalence varied with previous exposure; the prevalence of cLIA results positive for HPV16 and HPV18 was lower than the prevalence of positive VLP ELISA responses. cLIA and VLP ELISA positivity predicted protection from subsequent infections with concordant types. The combined odds ratio for HPV16 and HPV18 cLIA positivity was 0.41 (95% confidence interval [CI], 0.21-0.80), and the combined odds ratio for the HPV16 and HPV18 VLP ELISA positivity was 0.65 (95% CI, 0.46-0.93). Of individual types, statistical significance was only reached for HPV16 cLIA positivity (odds ratio, 0.44; 95% CI, 0.15-0.94). CONCLUSIONS: Both assays showed an association between positive results and significant protection from subsequent infections for HPV16 and HPV18 combined. cLIA seroprevalence was lower than VLP ELISA, suggesting that the assay detects a subset of antibodies following natural infection that are specifically linked to immunity against subsequent HPV infection. PMID- 21628664 TI - Progressive multifocal leukoencephalopathy (PML) development is associated with mutations in JC virus capsid protein VP1 that change its receptor specificity. AB - Progressive multifocal leukoencephalopathy (PML), a fatal demyelinating disease caused by JC virus (JCV) infection of oligodendrocytes, may develop in patients with immune disorders following reactivation of chronic benign infection. Mutations of JCV capsid viral protein 1 (VP1), the capsid protein involved in binding to sialic acid cell receptors, might favor PML onset. Cerebrospinal fluid sequences from 37/40 PML patients contained one of several JCV VP1 amino acid mutations, which were also present in paired plasma but not urine sequences despite the same viral genetic background. VP1-derived virus-like particles (VLPs) carrying these mutations lost hemagglutination ability, showed different ganglioside specificity, and abolished binding to different peripheral cell types compared with wild-type VLPs. However, mutants still bound brain-derived cells, and binding was not affected by sialic acid removal by neuraminidase. JCV VP1 substitutions are acquired intrapatient and might favor JCV brain invasion through abrogation of sialic acid binding with peripheral cells, while maintaining sialic acid-independent binding with brain cells. PMID- 21628665 TI - The acute phase of Chikungunya virus infection in humans is associated with strong innate immunity and T CD8 cell activation. AB - BACKGROUND: Rapidly spreading to new regions, including the islands of the Indian Ocean, Central Africa, and Europe, Chikungunya fever is becoming a major problem of public health. Unlike other members of the alphavirus genus, immune responses to Chikungunya virus (CHIKV) have been poorly investigated. METHODS: We conducted a large ex vivo multiplex study of 50 cytokine, chemokine, and growth factor plasma profiles in 69 acutely infected patients from the Gabonese outbreak of 2007. We also assessed a phenotypic study of T lymphocyte responses during human acute CHIKV infection. RESULTS: CHIKV infection in humans elicited strong innate immunity involving the production of numerous proinflammatory mediators. Interestingly, high levels of Interferon (IFN) alpha were consistently found. Production of interleukin (IL) 4, IL-10, and IFN-gamma suggested the engagement of the adaptive immunity. This was confirmed by flow cytometry of circulating T lymphocytes that showed a CD8+ T lymphocyte response in the early stages of the disease, and a CD4+ T lymphocyte mediated response in the later stages. For the first time to our knowledge, we found evidence of CD95-mediated apoptosis of CD4+ T lymphocytes during the first 2 days after symptoms onset, ex vivo. CONCLUSIONS: Together, our findings suggest that strong innate immunity is required to control CHIKV infection. PMID- 21628667 TI - Relationship between residual plasma viremia and the size of HIV proviral DNA reservoirs in infected individuals receiving effective antiretroviral therapy. AB - Residual plasma viremia (<50 copies/mL) persists in certain human immunodeficiency virus (HIV)-infected individuals receiving antiretroviral therapy (ART); however, the relationship between the degree of residual plasma viremia, the size of HIV reservoirs, and the level of immune activation has not been delineated. Here, we demonstrate that residual plasma viremia correlates with the size of the CD4(+) T cell viral reservoir, but not with markers of immune activation, suggesting that reactivation of the latent viral reservoir may not be the sole source of residual plasma viremia. Novel therapeutic strategies aimed at targeting the source of residual viremia may be necessary to achieve viral eradication. PMID- 21628666 TI - Safety and immunogenicity of a monovalent 2009 influenza A/H1N1v vaccine adjuvanted with AS03A or unadjuvanted in HIV-infected adults: a randomized, controlled trial. AB - BACKGROUND: Human immunodeficiency virus (HIV)-infected patients have decreased immune response to vaccines. Few data are available about pandemic flu vaccination in this population. METHODS: We conducted a multicenter, patient blinded, randomized trial in a cohort of HIV-infected adults. Patients received 2 injections 21 days apart of a AS03(A)-adjuvanted H1N1v vaccine containing 3.75 MUg hemagglutinin (HA) or a nonadjuvanted H1N1v vaccine containing 15 MUg HA to assess hemagglutination inhibition (HI) response and safety. RESULTS: A total of 309 patients were randomized, and 306 were vaccinated. After the first vaccine dose, HI titers >=1:40 were observed in 93.4% of the patients in the adjuvanted group (A group) (n = 155) and in 75.5% in the nonadjuvanted group (B group) (n = 151) (P < .001); seroconversion rates were 88.8% and 71.2%, and factor increases in geometric mean titers (GMT) of 21.9 and 15.1, respectively. After 2 injections, 98.6% of patients of the A group and 92.1% of the B group demonstrated HI titers >=1:40 (P = .018); seroconversion rates were 96.5% and 87.1%, respectively, and factor increases in GMT were 45.5 and 21.2, respectively. The majority of adverse events were mild to moderate in severity; no impact on CD4+ cell count or viral load has been detected. CONCLUSIONS: In HIV 1-infected adults, the AS03(A)-adjuvanted H1N1v vaccine yielded a higher immune response than did the nonadjuvanted one, with no impact on HIV infection. PMID- 21628668 TI - Impact of baseline HIV-1 tropism on viral response and CD4 cell count gains in HIV-infected patients receiving first-line antiretroviral therapy. AB - BACKGROUND: Viral tropism influences the natural history of human immunodeficiency type 1 (HIV-1) disease: X4 viruses are associated with faster decreases in CD4 cell count. There is scarce information about the influence of viral tropism on treatment outcomes. METHODS: Baseline plasma samples from patients recruited to the ArTEN (Atazanavir/ritnoavir vs. Nevirapine on a background of Tenofovir and Emtricitabine) trial were retrospectively tested for HIV-1 tropism using the genotypic tool geno2pheno(FPR=5.75%). ArTEN compared nevirapine with atazanavir-ritonavir, both along with tenofovir-emtricitabine, in drug-naive patients. RESULTS: Of 569 ArTEN patients, 428 completed 48 weeks of therapy; 282 of these received nevirapine and 146 of these received atazanavir ritonavir. Overall, non-B subtypes of HIV-1 were recognized in 96 patients (22%) and X4 viruses were detected in 55 patients (14%). At baseline, patients with X4 viruses had higher plasma HIV RNA levels (5.4 vs 5.2 log copies/mL, respectively; P = .044) and lower CD4 cell counts (145 vs 188 cells/MUL, respectively; P < .001) than those with R5 strains. At week 48, virologic responses were lower in patients with X4 viruses than in patients with R5 viruses (77% vs 92%, respectively; P = .009). Multivariate analysis confirmed HIV-1 tropism as an independent predictor of virologic response at week 24 (P = .012). This association was extended to week 48 (P = .007) in clade B viruses. Conversely, CD4 cell count recovery was not influenced by baseline HIV-1 tropism. CONCLUSIONS: HIV-1 tropism is an independent predictor of virologic response to first-line antiretroviral therapy. In contrast, it does not seem to influence CD4 cell count recovery. CLINICAL TRIALS REGISTRATION: NCT00389207. PMID- 21628669 TI - Genetic variants of ABCC10, a novel tenofovir transporter, are associated with kidney tubular dysfunction. AB - BACKGROUND: Tenofovir (TFV) causes kidney tubular dysfunction (KTD) in some patients, but the mechanism is poorly understood. Genetic variants in TFV transporters are implicated; we explored whether ABCC10 transports TFV and whether ABCC10 single-nucleotide polymorphisms (SNPs) are associated with KTD. METHODS: TFV accumulation was assessed in parental and ABCC10-transfected HEK293 cells (HEK293-ABCC10), CD4(+) cells and monocyte-derived macrophages (MDMs). Substrate specificity was confirmed by cepharanthine (ABCC10 inhibitor) and small interfering RNA (siRNA) studies. Fourteen SNPs in ABCC10 were genotyped in human immunodeficiency virus-positive patients with KTD (n = 19) or without KTD (controls; n = 96). SNP and haplotype analysis was performed using Haploview. RESULTS: TFV accumulation was significantly lower in HEK293-ABCC10 cell lines than in parental HEK293 cells (35% lower; P = .02); this was reversed by cepharanthine. siRNA knockdown of ABCC10 resulted in increased accumulation of TFV in CD4(+) cells (18%; P = .04) and MDMs (25%; P = .04). Two ABCC10 SNPs (rs9349256: odds ratio [OR], 2.3; P = .02; rs2125739, OR, 2.0; P = .05) and their haplotype (OR, 2.1; P = .05) were significantly associated with KTD. rs9349256 was associated with urine phosphorus wasting (P = .02) and beta2 microglobulinuria (P = .04). CONCLUSIONS: TFV is a substrate for ABCC10, and genetic variability within the ABCC10 gene may influence TFV renal tubular transport and contribute to the development of KTD. These results need to be replicated in other cohorts. PMID- 21628670 TI - Soluble CD163 made by monocyte/macrophages is a novel marker of HIV activity in early and chronic infection prior to and after anti-retroviral therapy. AB - CD163, a monocyte- and macrophage-specific scavenger receptor, is shed during activation as soluble CD163 (sCD163). We have previously demonstrated that monocyte expansion from bone marrow with simian immunodeficiency virus (SIV) infection correlated with plasma sCD163, the rate of AIDS progression, and the severity of macrophage-mediated pathogenesis. Here, we examined sCD163 in human immunodeficiency virus (HIV) infection. sCD163 was elevated in the plasma of individuals with chronic HIV infection (>1 year in duration), compared with HIV seronegative individuals. With effective antiretroviral therapy (ART), sCD163 levels decreased in parallel with HIV RNA levels but did not return to HIV seronegative levels, suggesting the presence of residual monocyte/macrophage activation even with plasma viral loads below the limit of detection. In individuals with early HIV infection (<=1 year in duration), effective ART resulted in decreased sCD163 levels that were comparable to levels in HIV seronegative individuals. sCD163 levels in plasma were positively correlated with the percentage of CD14+CD16+ monocytes and activated CD8+HLA-DR+CD38+ T lymphocytes and were inversely correlated with CD163 expression on CD14+CD16+ monocytes. With ART interruption in subjects with early HIV infection, sCD163 and plasma virus levels spiked but rapidly returned to baseline with reinitiation of ART. This study points to the utility of monocyte- and macrophage-derived sCD163 as a marker of HIV activity that links viral replication with monocyte and macrophage activation. These observations underscore the significance of monocyte and macrophage immune responses with HIV pathogenesis. PMID- 21628673 TI - Possibility of HIV-1 resistance mutations in cerebrospinal fluid from persons receiving suppressive therapy. PMID- 21628671 TI - Prevention of infection by a granulocyte-macrophage colony-stimulating factor co expressing DNA/modified vaccinia Ankara simian immunodeficiency virus vaccine. AB - A simian immunodeficiency virus (SIV) vaccine coexpressing granulocyte-macrophage colony stimulating factor (GM-CSF) prevented infection in 71% of macaques that received 12 rectal challenges. The SIVsmE660 challenge had the tropism of incident human immunodeficiency virus (HIV) infections and a similar genetic distance from the SIV239 vaccine as intraclade HIV isolates. The heterologous prime-boost vaccine regimen used recombinant DNA for priming and recombinant modified vaccinia Ankara for boosting. Co-expression of GM-CSF in the DNA prime enhanced the avidity of elicited immunoglobulin G for SIV envelope glycoproteins, the titers of neutralizing antibody for easy-to-neutralize SIV isolates, and antibody-dependent cellular cytotoxicity. Impressively, the co-expressed GM-CSF increased vaccine-induced prevention of infection from 25% in the non-GM-CSF co expressing vaccine group to 71% in the GM-CSF co-expressing vaccine group. The prevention of infection showed a strong correlation with the avidity of the elicited Env-specific antibody for the Env of the SIVsmE660 challenge virus (r = 0.9; P < .0001). PMID- 21628675 TI - Unconscious high-level information processing: implication for neurobiological theories of consciousness. AB - Theories about the neural correlates and functional relevance of consciousness have traditionally assigned a crucial role to the prefrontal cortex in generating consciousness as well as in orchestrating high-level conscious control over behavior. However, recent neuroscientific findings show that prefrontal cortex can be activated unconsciously. The depth, direction, and scope of these activations depend on several top-down factors such as the task being probed (task-set, strategy) and on (temporal/spatial) attention. Regardless, such activations-when mediated by feedforward activation only-do not lead to a conscious sensation. Although unconscious, these prefrontal activations are functional, in the sense that they are associated with behavioral effects of cognitive control, such as response inhibition, task switching, conflict monitoring, and error detection. These findings challenge the pivotal role of the prefrontal cortex in consciousness. Instead, it appears that specific brain areas (or cognitive modules) may support specific cognitive functions but that consciousness is independent of this. Conscious sensations arise only when the brain areas involved engage in recurrent interactions enabling the long-lasting exchange of information between brain regions. Moreover, recent evidence suggests that also the state of consciousness, for example, in vegetative state patients or during sleep and anesthesia, is closely related to the scope and extent of residual recurrent interactions among brain regions. PMID- 21628676 TI - The alpha (alpha)-glucosidase inhibitor, acarbose, attenuates the blood pressure and splanchnic blood flow responses to intraduodenal sucrose in older adults. AB - BACKGROUND: Postprandial hypotension is an important problem in the elderly and may be triggered by the increase in splanchnic blood flow induced by a meal. Acarbose attenuates the fall in blood pressure (BP) induced by oral sucrose and may be useful in the management of postprandial hypotension. It is not known whether the effect of acarbose on postprandial BP reflects slowing of gastric emptying and/or carbohydrate absorption nor whether acarbose affects splanchnic blood flow. We examined the effects of intraduodenal (ID) acarbose on the BP, heart rate, superior mesenteric artery (SMA) flow, and glycemic and insulin responses to ID sucrose in older participants--this approach excluded any "gastric" effect of acarbose. METHODS: Eight healthy participants (four male and four female, age 66-77 years) received an ID infusion of sucrose (~6 kcal/min), with or without acarbose (100 mg), over 60 minutes. BP, heart rate, SMA flow, blood glucose, and serum insulin were measured. RESULTS: Acarbose markedly attenuated the falls in systolic (p < .01) and diastolic (p < .05) BP and rises in heart rate (p < .05), SMA flow (p < .05), blood glucose (p < .01), and serum insulin (p < .05). The maximum fall in systolic BP and peak SMA flow was inversely related on the control day (r(2) = -.53, p < .05) but not with acarbose (r(2) = .03, p = .70). CONCLUSIONS: We conclude that in healthy older participants receiving ID sucrose, (a) acarbose markedly attenuates the hypotensive response by slowing carbohydrate absorption and attenuating the rise in splanchnic blood flow and (b) the fall in BP is related to the concomitant increase in SMA flow. PMID- 21628677 TI - Predictors of thyroid hormone initiation in older adults: results from the cardiovascular health study. AB - BACKGROUND: Despite widespread use, there are no data on initiation of thyroid hormone use in older people. We report the prevalence of thyroid hormone use and predictors of thyroid hormone initiation in a population of older men and women. METHODS: Thyroid hormone medication data were collected annually from 1989 to 2006 in community-dwelling individuals aged 65 years and older enrolled in the Cardiovascular Health Study (N = 5,888). Associations of age, sex, race, body mass index, education, and coronary heart disease with initiation were evaluated using discrete-time survival analysis. RESULTS: In 1989-1990, 8.9% (95% confidence interval 8.1%-9.7%) of participants were taking a thyroid hormone preparation, increasing to 20.0% (95% confidence interval 8.2%-21.8%) over 16 years. The average initiation rate was 1% per year. The initiation rate was nonlinear with age, and those aged 85 years and older initiated thyroid hormone more than twice as frequently as those aged 65-69 years (hazard ratio = 2.34; 95% confidence interval 1.43-3.85). White women were more likely to initiate thyroid hormone than any other race and sex group. Higher body mass index was independently associated with higher risk for initiation (p = .002) as was greater education (p = .02) and prevalent coronary heart disease (p = .03). CONCLUSIONS: Thyroid hormone use is common in older people. The indications and benefits of thyroid hormone use in older individuals with the highest rate of thyroid hormone initiation-the oldest old, overweight and obese individuals, and those with coronary heart disease-should be investigated. PMID- 21628679 TI - Denture care. PMID- 21628678 TI - Tobacco-use treatment in dental practice: how Healthy People 2020 aligns federal policy with the evidence. PMID- 21628680 TI - Acetaminophen use. PMID- 21628681 TI - Evidence-based dentistry recommendations. PMID- 21628682 TI - Minimally invasive management of dental caries: contemporary teaching of posterior resin-based composite placement in U.S. and Canadian dental schools. AB - BACKGROUND: Resin-based composites are an increasingly popular material for restoring posterior teeth, permitting minimally invasive cavity preparations and esthetic restorations. The authors investigated current teaching of the placement of posterior resin-based composites in U.S. and Canadian dental schools. METHODS: In late 2009 and early 2010, the authors, with the assistance of the Consortium of Operative Dentistry Educators (CODE), invited 67 dental schools to participate in an Internet-based survey. RESULTS: The response rate was 73 percent. Although all schools taught the placement of resin-based composites in occlusal and most occlusoproximal cavities, eight schools (16 percent) did not teach placement of three-surface occlusoproximal resin-based composite restorations in permanent molars. Resin-based composites accounted for 49 percent of direct posterior restorations placed by dental students in 2009 and 2010, a 30 percent increase from 2005. CONCLUSIONS: Teaching placement of posterior resin-based composites continues to increase in dental schools in the United States and Canada, with predoctoral students gaining, on average, an equal amount of experience placing posterior resin-based composites and amalgams in terms of numbers of restorations. CLINICAL IMPLICATIONS: Evidence-based, up-to-date teaching programs, including those in operative dentistry, are needed to best prepare students for careers in dentistry. PMID- 21628683 TI - Practitioner, patient and carious lesion characteristics associated with type of restorative material: findings from The Dental Practice-Based Research Network. AB - BACKGROUND: The authors conducted a study to identify factors associated with the materials that dentists in The Dental Practice-Based Research Network (DPBRN) use when placing the first restoration on permanent posterior tooth surfaces. METHODS: A total of 182 DPBRN practitioner-investigators provided data regarding 5,599 posterior teeth with caries. Practitioner-investigators completed an enrollment questionnaire that included the dentist's age, sex, practice workload, practice type and number of years since graduation. When patients who had provided informed consent to participate in the investigation sought treatment for a previously unrestored carious surface, the practitioner-investigator recorded patient and tooth characteristics. RESULTS: Practitioner-investigators used amalgam more often than they used direct resin-based composite (RBC) for posterior carious lesions. Practitioner and practice characteristics (years since graduation and type of practice); patient characteristics (sex, race, age and dental insurance status); and lesion characteristics (tooth location and surface, preoperative and postoperative lesion depth) were associated with the type of restorative material used. CONCLUSIONS: Several practitioner and practice, patient and lesion characteristics were associated significantly with use of amalgam and RBC: geographical region, years since dentist's graduation, patient's dental insurance status, tooth location and surface, and preoperative and postoperative lesion depth. CLINICAL IMPLICATIONS: Despite advances in esthetic dentistry, U.S. dentists still are placing amalgam on posterior teeth with carious lesions. Amalgam was used more often than RBC in older patients, who may have had deeper carious lesions. PMID- 21628684 TI - Laypeople's preferences regarding frontal dentofacial esthetics: tooth-related factors. AB - BACKGROUND: Researchers have conducted extensive studies regarding dentoalveolar factors that affect anterior dental esthetics; however, there is no consensus regarding laypeople's perspectives on these factors. METHODS: The authors conducted a systematic search of electronic databases (MEDLINE, PubMed, Embase, Cochrane Library and Web of Science) until May 2010. They identified and selected articles in which investigators explored anterior dental esthetics from a layperson's perspective, and they assigned methodological scores to the studies. RESULTS: Seventeen articles met the inclusion criteria. The authors determined laypeople's preferences for tooth shape, tooth size and proportion, and incisor position. CONCLUSIONS: The results of this literature review show that laypeople did not discriminate between square, square-round (basically square with rounded mesioincisal and distoincisal angles) and round incisors or between canine shapes when displayed in photographs of female models. They preferred square-round incisors to square incisors and flat canines (when paired with round incisors) in photographs of male models. Most laypeople appeared to prefer unworn dentitions, small teeth in photographs of female models and large teeth in photographs of male models, width-to-length ratios in central incisors between 75 and 85 percent, and tooth-to-tooth proportions between the lateral and central incisors between 50 and 74 percent. Laypeople discerned a 10 degrees angulation of one or both central incisors as being less attractive. Significant discrepancies in perceptions existed for incisal edges. Most laypeople preferred an overbite of 2.0 millimeters, with some leeway (around 2 mm). The authors noted that laypeople had a preference for no diastemas. CLINICAL IMPLICATIONS: Laypeople have varying degrees of sensitivity to certain esthetic issues. Thus, clinicians can expect their patients to be more attentive to some dental esthetic factors than to others. PMID- 21628685 TI - Treatment of complicated crown-root fracture in a single visit by means of rebonding. AB - BACKGROUND: Crown-root fractures of anterior teeth are relatively common but often time consuming and difficult to treat. In some cases, the clinician can complete the treatment in a single visit by reattaching the fractured fragment. CASE DESCRIPTION: In this case report, the authors describe the treatment of a complicated crown-root fracture of the maxillary right central and lateral incisors. Among the treatment options for such cases, the treating clinician considered a single-visit rebonding procedure to be the best choice. The treatment consisted of a multidisciplinary approach involving coronal fragment removal, gingivectomy to expose the margins and single-visit endodontic therapy, followed by a rebonding of the fractured fragment by means of prefabricated posts. RESULTS: Evaluation at 12 months indicated stable re-attachment of the fragments, good esthetics and good periodontal health. CLINICAL IMPLICATIONS: The reattachment of a tooth fragment is a viable, conservative technique that restores function and esthetics, and clinicians should consider it when treating patients with coronal fractures of the anterior teeth. PMID- 21628686 TI - Neuralgia-inducing cavitational osteonecrosis: a possible diagnosis for an orofacial pain complaint? PMID- 21628687 TI - Dietary Guidelines for Americans, 2010: an update. AB - BACKGROUND: The U.S. Department of Agriculture developed Dietary Guidelines for Americans, 2010, by taking into consideration evidence-based nutritional science, changes in the food supply and environmental influences. CONCLUSIONS: The 2010 guidelines emphasize the importance of maintaining caloric balance to support a healthy weight and selecting nutrient-dense foods and beverages to ensure adequate nutrient intakes. Americans' compliance with previous dietary guidelines has been poor, and the 2010 guidelines acknowledge the need for collaboration among individuals, the community, government and industry to ensure consumers' access to and acceptance of appropriate foods. CLINICAL IMPLICATIONS: As part of the greater community, the oral health practitioner will find that familiarity with the 2010 guidelines will enable him or her to assist patients in making better food choices for oral and systemic health. PMID- 21628688 TI - The color differences of direct esthetic restorative materials after setting and compared with a shade guide. AB - BACKGROUND: The authors conducted a study to evaluate esthetic restorative materials' color differences after setting and color matching between set materials and a shade guide. MATERIALS AND METHODS: The authors evaluated 13 resin-based composites, one silorane-based composite, two polyacid-modified resin composites and one conventional glass ionomer cement. They measured the color parameters of the samples, which were 8 millimeters in diameter and 1.5 mm in thickness, before and after they were set according to the Commission International de l'Eclairage (CIE) L*a*b* color scale relative to standard illumination against a white background by means of a dental colorimeter. They also compared the final colors of the restorative materials with a shade guide. RESULTS: Color difference values for each restorative material ranged from 3.25 to 14.04. With the exception of Fuji IX (GC, Tokyo), Filtek P60 (3M ESPE) and Te Econom (Ivoclar Vivadent), the restorative materials exhibited a perceptible color change after setting. Color difference values between the set materials and the shade guide tabs ranged from 1.86 to 11.83. With the exception of Filtek Supreme XT (3M ESPE) and Fuji IX, the materials exhibited a perceptible difference. CONCLUSION: Most of the materials tested exhibited a significant color change after polymerization and did not match the shade guide tab after undergoing light curing. PMID- 21628689 TI - A lack of consensus in the measurement methods for and definition of periodontitis. PMID- 21628690 TI - The all-ceramic restoration dilemma: where are we? PMID- 21628691 TI - Making the right investment decisions for the practice. PMID- 21628692 TI - What is my ethical responsibility when my employer authorizes treatments that I believe are unnecessary? PMID- 21628693 TI - Potential eligibility for recombinant tissue plasminogen activator therapy in children: a population-based study. AB - Intravenous recombinant tissue plasminogen activator is an established therapy for adults with ischemic stroke. In this Greater Cincinnati/Northern Kentucky population-based study, 8% were eligible. However, no established therapy exists for children with acute ischemic stroke. Accordingly, investigators assessed rates of eligibility for recombinant tissue plasminogen activator therapy among children (<18 years of age) in the same population to aid planning of future clinical trials. The investigators identified 29 pediatric ischemic strokes during 3 separate study periods (1993-1994, 1999, and 2005) and determined potential eligibility for recombinant tissue plasminogen activator therapy based on 2007 American Heart Association guidelines for adults. Depending on how relative contraindications were considered, 1 to 3 cases (3%-10%) met eligibility criteria. On the basis of national pediatric stroke incidence rates extrapolated from our population, it is estimated that up to 178 children might be eligible for intravenous recombinant tissue plasminogen activator therapy annually in the United States. Thus, recruitment for clinical studies is likely to be challenging and requires a concerted multicenter effort. PMID- 21628694 TI - Dynamic and structural formation of a thrombus: the inciting event of arterial ischemic stroke. AB - Arterial ischemic stroke occurs as a result of abnormal clinical circumstances that alter hemostasis and cause thrombosis, either within a vessel or as an embolic event. Understanding normal hemostasis, including differences between children (developmental hemostasis) and adults, will provide background for determining the pathophysiology of stroke and potential treatments. PMID- 21628695 TI - Cellular mechanisms of neurovascular damage and repair after stroke. AB - The biological processes underlying stroke are complex, and patients have a narrow repertoire of therapeutic opportunities. After the National Institutes of Health (NIH) convened the Stroke Progress Review Group in 2001, stroke research shifted from having a purely neurocentric focus to adopting a more integrated view wherein dynamic interactions between all cell types contribute to function and dysfunction in the brain. This so-called "neurovascular unit" provides a conceptual framework that emphasizes cell-cell interactions between neuronal, glial, and vascular elements. Under normal conditions, signaling within the neurovascular unit helps maintain homeostasis. After stroke, cell-cell signaling is disturbed, leading to pathophysiology. More recently, emerging data now suggest that these cell-cell signaling mechanisms may also mediate parallel processes of neurovascular remodeling during stroke recovery. Because plasticity is a signature feature of the young and developing brain, these concepts may have special relevance to how the pediatric brain responds after stroke. PMID- 21628696 TI - A prospective outcome study of neonatal cerebral sinovenous thrombosis. AB - Neonatal cerebral sinovenous thrombosis is a frequent contributor to neonatal mortality and morbidity. Treatment is controversial, and reported clinical outcomes vary widely. Newborns with radiologically confirmed neonatal cerebral sinovenous thrombosis from 1992 to 2009 were prospectively followed in our Children's Stroke Clinic for standardized outcomes, including the Pediatric Stroke Outcome Measure. Outcomes were available in 90 of 104 (87%) neonates. Early outcomes included cerebral sinovenous thrombosis-associated death (5) and thrombus propagation (15 [6 associated with new venous infarcts]). Lack of anticoagulation predicted propagation (RR = 13; P = .0007). Complete thrombus recanalization occurred in 90% by 3 months. Late outcomes (median, 2.5 years) were epilepsy (15) and neurological disability (50), which included moderate severe language (43), sensorimotor (38), and cognitive/behavioral (24) deficits. Overall, 61% had poor outcome (death/any deficit). Concurrent neurological comorbidity at diagnosis (odds ratio = 2.8; P = .029) predicted poor outcome. Clinical trials are urgently needed to establish more effective treatment strategies. PMID- 21628697 TI - The role of aspirin in childhood tuberculous meningitis. AB - Arterial stroke is the main cause of poor outcome in childhood tuberculous meningitis. Aspirin has an antithrombotic action at low dose and anti-ischemic and anti-inflammatory properties, which are dose-related. The aim of the study was to explore the possible benefits of aspirin in children with tuberculous meningitis. A total of 146 consecutive children with a diagnosis of probable tuberculous meningitis were studied. Patients were randomized into 3 groups: (1) placebo group, (2) low-dose aspirin group, and (3) high-dose aspirin group. Twenty-nine additional patients who received aspirin before admission were excluded from the randomized study, but continued on low-dose aspirin. Aspirin, irrespective of dose, did not show any significant benefit regarding morbidity (hemiparesis and developmental outcome) and mortality. Aspirin was well tolerated, but 1 death was probably related to aspirin. The fact that the outcome of the high-dose aspirin group compared favorably with the other treatment groups despite younger age and more severe neurological involvement at baseline needs further investigation. PMID- 21628698 TI - Down syndrome and attention-deficit/hyperactivity disorder (ADHD). AB - Clinicians might minimize the prevalence of behavioral disorders among mentally retarded people. Decreased attention, hyperactivity, and impulsivity are frequently reported in children with Down syndrome, yet the exact prevalence of attention-deficit/hyperactivity disorder (ADHD) has not been clearly estimated in this population. The objective of this study was to estimate the prevalence of ADHD in children with Down syndrome and to emphasize the possible relationship between ADHD symptoms and the level of mental retardation and common medical comorbidity. In this study, the prevalence of ADHD among Down syndrome children was very high, reaching 43.9%. No significant correlation was found between ADHD symptoms and the level of mental retardation, but significant correlation was found with ophthalmologic problems. We conclude that children with Down syndrome are at increased risk for ADHD. When evaluating children with Down syndrome for attention deficits, psychiatric comorbidity as well as medical problems should be carefully taken into consideration. PMID- 21628699 TI - Increased regional fractional anisotropy in highly screened attention-deficit hyperactivity disorder (ADHD). AB - Diffusion tensor imaging data were collected at 3.0 Tesla from 16 children with attention-deficit hyperactivity disorder (ADHD) and 16 typically developing controls, ages 9 to 14 years. Fractional anisotropy images were calculated and normalized by linear transformation. Voxel-wise and atlas-based region-of interest analyses were performed. Using voxel-wise analysis, fractional anisotropy was found to be significantly increased in the attention-deficit hyperactivity disorder group in the right superior frontal gyrus and posterior thalamic radiation, and left dorsal posterior cingulate gyrus, lingual gyrus, and parahippocampal gyrus. No regions showed significantly decreased fractional anisotropy in attention-deficit hyperactivity disorder. Region-of-interest analysis revealed increased fractional anisotropy in the left sagittal stratum, that is, white matter that connects the temporal lobe to distant cortical regions. Only fractional anisotropy in the left sagittal stratum was significantly associated with attention-deficit hyperactivity disorder symptom severity. Several recent studies have reported pathological increases in fractional anisotropy in other conditions, highlighting the relevance of diffusion tensor imaging in identifying atypical white matter structure associated with neurodevelopmental processes. PMID- 21628700 TI - Preventive psychiatry: a paradigm to improve population mental health and well being. AB - The government's Public Health White Paper for England sets out a utopian vision of how to prevent and remedy mental health problems. The public health approach relies on primary prevention, promoting individual responsibilities and resilience, while also sustaining existing services and tackling inequalities. These ambitions are consistent with the preventive psychiatric paradigm, and with the best of evidence-based psychiatric practice. Although the evidence on cost effectiveness of public mental health interventions is growing, the challenge is to ensure that specialist knowledge informs policy, practice and research so that inequalities are not compounded. Specialist mental health professionals are needed to inform and lead public health reforms. PMID- 21628701 TI - Forensic psychiatry and public protection. AB - The prominence of risk in UK social and criminal justice policy creates opportunities, challenges and dangers for forensic psychiatry. The future standing of the specialty will depend not only on the practical utility of its responses to those opportunities and challenges, but also the ethical integrity of those responses. PMID- 21628702 TI - The truth about genetic variation in the serotonin transporter gene and response to stress and medication. AB - The question of whether a functional variant in the promoter of the serotonin transporter gene (5-HTTLPR) influences response to adversity and/or antidepressants has generated great interest and controversy. A review of the literature suggests that the issue is complicated by differences in methodology and sample ethnicity. When these confounders are accounted for, there probably is a real, if small, effect of 5-HTTLPR on response to both serotonin reuptake inhibitors and environmental adversity. PMID- 21628703 TI - Psychological treatments in intellectual disability: the challenges of building a good evidence base. AB - Psychological treatments are widely used for the management of mental health and behavioural problems in people with intellectual disabilities. The evidence base, including the cost-effectiveness of such interventions, is limited. This editorial explores the current evidence base and analyses its strengths and limitations. The editorial also highlights current problems in conducting randomised controlled trials in this area and suggests a way forward. PMID- 21628704 TI - Dangerous and severe personality disorder. AB - The Dangerous and Severe Personality Disorder (DSPD) initiative was introduced a decade ago against overwhelming opposition from psychiatrists and others concerned with the implications of extending the public protection agenda through the use of a questionable medical 'diagnosis'. As this initiative is now being scaled down, it offers an opportunity to consider the positive and negative aspects of the initiative together with its longer-term legacy. PMID- 21628705 TI - Revascularisation and mortality rates following acute coronary syndromes in people with severe mental illness: comparative meta-analysis. AB - BACKGROUND: High levels of comorbid physical illness and excess mortality rates have been previously documented in people with severe mental illness, but outcomes following myocardial infarction and other acute coronary syndromes are less clear. AIMS: To examine inequalities in the provision of invasive coronary procedures (revascularisation, angiography, angioplasty and bypass grafting) and subsequent mortality in people with mental illness and in those with schizophrenia, compared with those without mental ill health. METHOD: Systematic search and random effects meta-analysis were used according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Studies of mental health and cardiovascular procedures following cardiac events were eligible but we required a minimum of three independent studies to warrant pooling by procedure type. We searched Medline/PubMed and EMBASE abstract databases and ScienceDirect, Ingenta Select, SpringerLink and Online Wiley Library full text databases. RESULTS: We identified 22 analyses of possible inequalities in coronary procedures in those with defined mental disorder, of which 10 also reported results in schizophrenia or related psychosis. All studies following acute coronary syndrome originated in the USA. The total sample size was 825 754 individuals. Those with mental disorders received 0.86 (relative risk, RR: 95% CI 0.80-0.92, P<0.0001) of comparable procedures with significantly lower receipt of coronary artery bypass graft (CABG; RR = 0.85, 95% CI 0.72 1.00), cardiac catheterisation (RR = 0.85, 95% CI 0.76-0.95) and percutaneous transluminal coronary angioplasty or percutaneous coronary intervention (PTCA/PCI; RR = 0.87, 95% CI 0.72-1.05). People with a diagnosis of schizophrenia received only 0.53 (95% CI 0.44-0.64, P<0.0001) of the usual procedure rate with significantly lower receipt of CABG (RR = 0.69, 95% CI 0.55-0.85) and PTCA/PCI (RR = 0.50, 95% CI 0.34-0.75). We identified 6 related studies examining mortality following cardiac events: for those with mental illness there was a 1.11 relative risk of mortality up to 1 year (95% CI 1.00-1.24, P = 0.05) but there was insufficient evidence to examine mortality rates in schizophrenia alone. CONCLUSIONS: Following cardiac events, individuals with mental illness experience a 14% lower rate of invasive coronary interventions (47% in the case of schizophrenia) and they have an 11% increased mortality rate. Further work is required to explore whether these factors are causally linked and whether improvements in medical care might improve survival in those with mental ill health. PMID- 21628706 TI - Cannabis use before age 15 and subsequent executive functioning. AB - BACKGROUND: Many studies have suggested that adolescence is a period of particular vulnerability to neurocognitive effects associated with substance misuse. However, few large studies have measured differences in cognitive performance between chronic cannabis users who started in early adolescence (before age 15) with those who started later. AIMS: To examine the executive functioning of individuals who started chronic cannabis use before age 15 compared with those who started chronic cannabis use after 15 and controls. METHOD: We evaluated the performance of 104 chronic cannabis users (49 early onset users and 55 late-onset users) and 44 controls who undertook neuropsychological tasks, with a focus on executive functioning. Comparisons involving neuropsychological measures were performed using generalised linear model analysis of variance (ANOVA). RESULTS: The early-onset group showed significantly poorer performance compared with the controls and the late-onset group on tasks assessing sustained attention, impulse control and executive functioning. CONCLUSIONS: Early-onset chronic cannabis users exhibited poorer cognitive performance than controls and late-onset users in executive functioning. Chronic cannabis use, when started before age 15, may have more deleterious effects on neurocognitive functioning. PMID- 21628707 TI - Grey matter and social functioning correlates of glutamatergic metabolite loss in schizophrenia. AB - BACKGROUND: Thalamic glutamine loss and grey matter reduction suggest neurodegeneration in first-episode schizophrenia, but the duration is unknown. AIMS: To observe glutamine and glutamate levels, grey matter volumes and social functioning in patients with schizophrenia followed to 80 months after diagnosis. METHOD: Grey matter volumes and proton magnetic resonance spectroscopy metabolites in left anterior cingulate and left thalamus were measured in 17 patients with schizophrenia before medication and 10 and 80 months after diagnosis. Social functioning was assessed with the Life Skills Profile Rating Scale (LSPRS) at 80 months. RESULTS: The sum of thalamic glutamate and glutamine levels decreased over 80 months, and correlated inversely with the LSPRS. Thalamic glutamine and grey matter loss were significantly correlated in frontal, parietal, temporal and limbic regions. CONCLUSIONS: Brain metabolite loss is correlated with deteriorated social functioning and grey matter losses in schizophrenia, consistent with neurodegeneration. PMID- 21628709 TI - Mixed anxiety and depressive disorder outcomes: prospective cohort study in primary care. AB - BACKGROUND: Mixed anxiety and depressive disorder (MADD) is common yet ill defined, with little known about outcomes. AIMS: To determine MADD outcomes over 1 year. METHOD: We recruited 250 adults attending seven London general practices with mild-moderate distress. Three groups were defined using a diagnostic interview: MADD, other ICD-10 psychiatric diagnosis, no psychiatric diagnosis. We assessed symptoms of distress (General Health Questionnaire-28), quality of life (12-item Short Form Health Survey), general practitioner (GP) diagnosis and consultation rate at baseline, 3 months and 1 year. RESULTS: Two-thirds of participants with MADD had no significant psychological distress at 3 months (61%) or 1 year (69%). However, compared with those with no diagnosis, individuals had twice the risk of significant distress (incidence rate ratio 2.39, 95% CI 1.29-4.42) at 3 months but not 1 year, and persistently lower quality of life (mental health functioning). There was no significant difference in GP consultation rate/diagnosis. CONCLUSIONS: The majority with MADD improved, but individuals had an increased risk of significant distress at 3 months and a lower quality of life. As we cannot currently predict those with a poorer prognosis these patients should be actively monitored in primary care. PMID- 21628708 TI - MAOA, abuse exposure and antisocial behaviour: 30-year longitudinal study. AB - BACKGROUND: Recent studies have raised issues concerning the replicability of gene * environment (G * E) interactions involving the monoamine oxidase A (MAOA) gene in moderating the associations between abuse or maltreatment exposure and antisocial behaviour. This study attempted to replicate the findings in this area using a 30-year longitudinal study that has strong resemblance to the original research cohort. AIMS: To test the hypothesis that the presence of the low activity MAOA genotype was associated with an increased response to abuse exposure. METHOD: Participants were 398 males from the Christchurch Health and Development Study who had complete data on: MAOA promoter region variable number tandem repeat genotype; antisocial behaviour to age 30; and exposure to childhood sexual and physical abuse. RESULTS: Regression models were fitted to five antisocial behaviour outcomes (self-reported property offending; self-reported violent offending; convictions for property/violent offending; conduct problems; hostility) observed from age 16 to 30, using measures of childhood exposure to sexual and physical abuse. The analyses revealed consistent evidence of G * E interactions, with those having the low-activity MAOA variant and who were exposed to abuse in childhood being significantly more likely to report later offending, conduct problems and hostility. These interactions remained statistically significant after control for a range of potentially confounding factors. Findings for convictions data were somewhat weaker. CONCLUSIONS: The present findings add to the evidence suggesting that there is a stable G * E interaction involving MAOA, abuse exposure and antisocial behaviour across the life course. PMID- 21628710 TI - Age and birth cohort differences in the prevalence of common mental disorder in England: National Psychiatric Morbidity Surveys 1993-2007. AB - BACKGROUND: There are concerns that the prevalence of mental disorder is increasing. AIMS: To determine whether the prevalence of common adult mental disorders has increased over time, using age-period-cohort analysis. METHOD: The study consisted of a pseudocohort analysis of a sequence of three cross-sectional surveys of the English household population. The main outcome was common mental disorder, indicated by a score of 12 or above on the Revised Clinical Interview Schedule (CIS-R). Secondary outcomes were neurotic symptoms likely to require treatment, indicated by a CIS-R score of 18 or over, and individual subscale scores for fatigue, sleep problems, irritability and worry. RESULTS: There were 8670 participants in the 1993 survey, 6977 in the 2000 survey and 6815 in the 2007 survey. In men a significant increase in common mental disorder occurred between the cohort born in 1943-9 and that born in 1950-6 (odds ratio 1.4, 95% CI 1.1-1.9) but prevalence in subsequent cohorts remained largely stable. More extended increases in prevalence of sleep problems and mental disorders were observed in women, but not consistently across cohorts or measures. CONCLUSIONS: We found little evidence that the prevalence of common mental disorder is increasing. PMID- 21628711 TI - Trends in rates of mental illness in homicide perpetrators. AB - BACKGROUND: The rise in homicides by those with serious mental illness is of concern, although this increase may not be continuing. AIMS: To examine rates of mental illness among homicide perpetrators. METHOD: A national consecutive case series of homicide perpetrators in England and Wales from 1997 to 2006. Rates of mental disorder were based on data from psychiatric reports, contact with psychiatric services, diminished responsibility verdict and hospital disposal. RESULTS: Of the 5884 homicides notified to the National Confidential Inquiry into Suicide and Homicide by People with Mental Illness between 1997 and 2006, the number of homicide perpetrators with schizophrenia increased at a rate of 4% per year, those with psychotic symptoms at the time of the offence increased by 6% per year. The number of verdicts of diminished responsibility decreased but no change was found in the number of perpetrators receiving a hospital order disposal. The likeliest explanation for the rise in homicide by people with psychosis is the misuse of drugs and/or alcohol, which our data show increased at a similar magnitude to homicides by those with psychotic symptoms. However, we are unable to demonstrate a causal association. Although the Poisson regression provides evidence of an upward trend in homicide by people with serious mental illness between 1997 and 2006, the number of homicides fell in the final 2 years of data collection, so these findings should be treated with caution. CONCLUSIONS: There appears to be a concomitant increase in drug misuse over the period, which may account for this rise in homicide. However, an increase in the number of people in contact with mental health services may suggest that access to mental health services is improving. Previous studies have used court verdicts such as diminished responsibility as a proxy measure of mental disorder. Our data indicate that this does not reflect accurately the prevalence of mental disorder in this population. PMID- 21628712 TI - Assisted suicide: two sides to the debate. PMID- 21628713 TI - Domestic violence: we need changes in the ICD and at the start of training. PMID- 21628714 TI - Lithium concentrations in drinking water. PMID- 21628715 TI - Suicide as protest against social suffering in the Arab world. PMID- 21628716 TI - Dose-related induction of hepatic preneoplastic lesions by diethylnitrosamine in C57BL/6 mice. AB - The C57BL/6 mouse strain (or derivation of this strain) is used as a background for many transgenic mouse models. This strain has a relatively low susceptibility to chemically induced hepatocarcinogenesis compared with other commonly used experimental mouse strains. In the present study, the authors treated C57BL/6 mice with 25, 50, and 75 mg/kg of diethylnitrosamine (DEN) for 4 or 8 weeks by intraperitoneal injection to investigate the dose-response pattern of preneoplastic and neoplastic lesion formation in the liver. DEN induced preneoplastic lesions and cytokeratin 8/18-positive foci in a dose-dependent manner. In the 75 mg/kg for 8 weeks treatment group, hepatocellular adenoma, cholangioma and hemangioma, and cytokeratin 19-positive foci were also induced, but a significant decrease in body weight was observed. The suitable DEN treatment range for this strain was concluded to be from 75 mg/kg for 4 weeks (total amount = 300 mg/kg) to 50 mg/kg for 8 weeks (total amount = 400 mg/kg). These results should prove useful for future studies investigating hepatocarcinogenesis in both the background C57BL/6 strain and other transgenic mouse models derived from it. PMID- 21628717 TI - Evaluation of the rabbit nasal cavity in inhalation studies and a comparison with other common laboratory species and man. AB - The rabbit is occasionally used for inhalation and intranasal safety assessment studies, but there are no detailed descriptions of the anatomy or histology of the rabbit nose. To address this deficit, the nasal cavities of thirty-two control adult rabbits were sectioned and examined to provide mapping of the main epithelial types and histological structures present within the cavity and turbinates. Four levels of the nasal cavity were prepared and examined using anatomic landmarks. Level I was sectioned immediately posterior to the incisors, Level II at the first palatal ridge, Level III immediately anterior to the first upper premolar teeth, and Level IV immediately anterior to the first upper molar. Level I was lined predominantly by squamous epithelium with small amounts of thick transitional epithelium, and examination is recommended only for studies involving test article administration via instillation. Level II was lined primarily with transitional and respiratory epithelia, whereas Levels III and IV were lined with respiratory and olfactory epithelia and often contained nasal associated lymphoid tissue. The vomeronasal organs were evident only in Level II. The similarities and differences of these features are compared with those of other common laboratory species (rat, mouse, dog, and cynomolgus monkey) and man. PMID- 21628718 TI - Safety evaluation of chronic intrathecal administration of idursulfase-IT in cynomolgus monkeys. AB - Recombinant human idursulfase, an intravenous enzyme replacement therapy indicated for treatment of somatic symptoms of mucopolysaccharidosis II (Hunter syndrome), is anticipated to have minimal benefit for the cognitive impairment associated with the severe phenotype. Because intrathecal (IT) administration of enzyme replacement therapy for other lysosomal enzyme disorders has shown efficacy in animal models, an IT formulation of idursulfase (idursulfase-IT) and a drug-delivery device (subcutaneous port connected to a lumbar IT catheter) were developed for treating central nervous system (CNS) involvement. In this chronic safety study, cynomolgus monkeys were dosed weekly with IV idursulfase (0.5 mg/kg) and every four weeks with idursulfase-IT (3, 30, and 100 mg) for six months, with device and vehicle controls treated similarly (n = 6, all groups). Necropsies were performed twenty-four hours post-final IT dose or after a recovery period (four weeks post-final dose in vehicle-control, 3 mg, and 100 mg IT groups: n = 6). No clinical signs or gross central nervous system lesions were observed. Compared to controls, more pronounced cellular infiltrates in brain and spinal cord meninges were noted, which largely resolved after the recovery period. Central nervous sytem levels of idursulfase-IT were dose dependent, as determined by enzyme activity and immunohistochemistry. The no-observed-adverse effect level of idursulfase-IT was 100 mg. PMID- 21628719 TI - Spontaneous glomerulonephritis in Gottingen minipigs. AB - The Gottingen minipig is one of the nonrodent species recommended by various regulatory authorities for safety assessment of drugs in preclinical studies. In such studies, knowledge of background pathology is critical in order to evaluate the potential renal toxicity. In the present study, the authors report 4 cases of glomerulonephritis out of 154 microbiologically defined Gottingen minipigs microscopically evaluated in preclinical studies. One animal required early sacrifice because of general poor health, and an additional animal died spontaneously. Histopathological evaluation revealed renal lesions in all 4 animals, exhibiting membranous or membranoproliferative glomerulonephritis at different stages, accompanied by secondary tubulo-interstitial damage. The renal changes observed were considered spontaneous in origin and of unknown etiology. Development of this condition in this strain should be considered in future studies. PMID- 21628720 TI - Right ventricular remodelling in systemic hypertension: a cardiac MRI study. AB - BACKGROUND: Consistent evidence shows an impact of systemic haemodynamic overload on the right ventricle, but its functional and structural consequences have received scarce attention for several reasons including the difficult application of conventional imaging techniques due to the complex shape and orientation of that cardiac chamber. AIMS: To evaluate whether mild to moderate, uncomplicated hypertension associates with abnormal right ventricular structure and function and how those changes relate to homologous changes in the left ventricle. Data were acquired by steady-state free-precession cardiac MRI, the state of the art tool for the morphological and functional evaluation of the right ventricle. MATERIALS AND METHODS: Twenty-five (12 women) uncomplicated, untreated, essential hypertensive patients were compared with 24 (13 women) sedentary normotensive controls of comparable age. Wall thickness, indexed ventricular mass, end diastolic volumes, early peak filling rate, a correlate of diastolic relaxation, and ejection fraction were measured at both ventricles. Remodelling index, the ratio of ventricular mass to end-diastolic volume, was used as an index of concentricity. RESULTS: Right ventricular mass index, ventricular wall thickness and remodelling index were greater in hypertensive subjects and associated with reduced peak filling rate, a pattern consistent with concentric right ventricular remodelling. In the hypertensive group, positive, highly significant biventricular correlations existed between indexed mass, early peak filling rate and ejection fraction. CONCLUSIONS: Systemic hypertension associates with concentric right ventricular remodelling and impaired diastolic function, confirming that the unstressed ventricle is not immune to the effects of systemic hypertension. Structural and functional right ventricular adaptation to systemic hypertension tends to parallel the homologous modifications induced by systemic haemodynamic overload on the left ventricle. PMID- 21628721 TI - Variability in on-treatment platelet reactivity explained by CYP2C19*2 genotype is modest in clopidogrel pretreated patients undergoing coronary stenting. AB - BACKGROUND: An inadequate response to clopidogrel is mainly attributable to the variable formation of its active metabolite. The CYP2C19*2 loss-of-function polymorphism leads to reduced generation of the active metabolite and is, similarly to high on-treatment platelet reactivity (HPR), associated with recurrent atherothrombotic events following coronary stent implantation. AIM: To determine the relative contribution of CYP2C19*2 genotype to HPR. METHODS AND RESULTS: CYP2C19*2 genotyping and platelet function testing using 5 and 20 MUmol/l ADP-induced light transmittance aggregometry (LTA), the PlateletWorks assay and the VerifyNow P2Y12 assay, were performed in 1069 clopidogrel pretreated patients undergoing elective coronary stenting (POPular study, http://clinicalTrials.gov/ NCT00352014). The relative contributions of CYP2C19*2 genotype and clinical variables to the interindividual variability of on treatment platelet reactivity and the occurrence of HPR were established using multivariate regression models. CYP2C19*2 carrier status was associated with a more frequent occurrence of HPR. CYP2C19*2 genotype alone could explain 5.0%, 6.2%, 4.4% and 3.7% of the variability in 5 and 20 MUmol/l ADP-induced LTA, the PlateletWorks assay and the VerifyNow P2Y12 assay, respectively, which increased to 13.0%, 15.2%, 5.6% and 20.6% when clinical variables were considered as well. Besides the CYP2C19*2 genotype, multiple clinical variables could be identified as independent predictors of HPR, including age, gender, body mass index, diabetes mellitus, clopidogrel loading dose regimen, use of amlodipine and platelet count. CONCLUSION: The CYP2C19*2 loss-of-function polymorphism is associated with a more frequent occurrence of HPR. However, the part of the interindividual variability in on-treatment platelet reactivity explained by CYP2C19*2 genotype is modest. PMID- 21628722 TI - Incremental prognostic value of early postoperative right ventricular systolic function in patients undergoing surgery for isolated severe tricuspid regurgitation. AB - BACKGROUND: Preoperative right ventricular end-systolic area (RV-ESA) and haemoglobin level have been suggested to be independent predictors of long-term prognosis in patients undergoing corrective surgery for isolated severe tricuspid regurgitation (TR). AIMS: To investigate whether early postoperative echocardiography provides useful prognostic information in addition to preoperative clinical and echocardiographic variables. METHODS: 69 consecutive patients undergoing corrective surgery for isolated severe TR (60 women, mean 57.6+/-8.9 years) were studied. Comprehensive preoperative echocardiography was performed in all patients, with early postoperative echocardiography in all patients except one. During follow-up (median 40 months, range 6-86 months), clinical events were investigated, defined as operative mortality, cardiovascular death, repeated open heart surgery and readmission due to cardiovascular problems. RESULTS: 28 patients (41%) were categorised as New York Heart Association class II, 36 (52%) as III and 5 (7%) as IV. 63 patients (91.3%) had undergone prior left-sided valve surgery. Seven (10.1%) patients died before discharge. Of the remaining 62 patients, three died during follow-up, and eight required readmission due to cardiovascular problems. On multivariate analyses using clinical and preoperative variables, RV-ESA (p=0.006) and haemoglobin level (p<0.001) were independent predictors of event-free survival. When early postoperative echocardiography variables were included, preoperative haemoglobin and early postoperative RV-FAC were predictors of long-term event-free survival. On receiver-operating characteristic curve analysis, early postoperative RV FAC>=31% predicted event-free survival with a sensitivity of 90% and a specificity of 83% (p<0.001). The addition of early postoperative echocardiographic RV-FAC markedly improved the prognostic utility of the model containing preoperative haemoglobin level and echocardiographic RV-ESA (p<0.001). CONCLUSION: Early postoperative RV-FAC, measured by echocardiography, provided valuable information additional to preoperative RV-ESA and hemoglobin level that was useful for predicting long-term clinical events following corrective TR surgery. PMID- 21628723 TI - Effects of aging on control of timing and force of finger tapping. AB - The present study examined whether the elderly produced a hastened or delayed tap with a negative or positive constant intertap interval error more frequently in self-paced tapping than in the stimulus-synchronized tapping for the 2 N target force at 2 or 4 Hz frequency. The analysis showed that, at both frequencies, the percentage of the delayed tap was larger in the self-paced tapping than in the stimulus-synchronized tapping, whereas the hastened tap showed the opposite result. At the 4 Hz frequency, all age groups had more variable intertap intervals during the self-paced tapping than during the stimulus-synchronized tapping, and the variability of the intertap intervals increased with age. Thus, although the increase in the frequency of delayed taps and variable intertap intervals in the self-paced tapping perhaps resulted from a dysfunction of movement timing in the basal ganglia with age, the decline in timing accuracy was somewhat improved by an auditory cue. The force variability of tapping at 4 Hz further increased with age, indicating an effect of aging on the control of force. PMID- 21628724 TI - Effect of coordination biofeedback on (re)learning preferred postural patterns in post-stroke patients. AB - After stroke, ankle-hip coordination during stance is characterized by changes in the postural system dynamics, specifically the disappearance of the in-phase pattern and the reduced stability of the anti-phase pattern. This study was conducted to assess the success of a coordination visual biofeedback for the (re)learning of the two preferred patterns, and to explore the effect of this treatment on postural and functional abilities. Twenty four patients were randomly assigned to one of two experimental groups or to a control group. During one month, patients from experimental groups followed a training protocol on the two preferred postural patterns using the biofeedback device. These two groups improved their in-phase coordination after the (re)learning compared with control group, and showed a related improvement of the functional independence measure. Results suggest that (re)learning the in-phase pattern is possible and seems to improve independence in poststroke patients. PMID- 21628725 TI - Arm motion coupling during locomotion-like actions: an experimental study and a dynamic model. AB - We studied the coordination of arm movements in standing persons who performed an out-of-phase arm-swinging task while stepping in place or while standing. The subjects were instructed to stop one of the arms in response to an auditory signal while trying to keep the rest of the movement pattern unchanged. A significant increase was observed in the amplitude of the arm that continued swinging under both the stepping and standing conditions. This increase was similar between the right and left arms. A dynamic model was developed including two coupled nonlinear van der Pol oscillators. We assumed that stopping an arm did not eliminate the coupling but introduced a new constraint. Within the model, superposition of two factors, a command to stop the ongoing movement of one arm and the coupling between the two oscillators, has been able to account for the observed effects. The model makes predictions for future experiments. PMID- 21628726 TI - Cardioballistic impulse and fluctuations in isometric force output. AB - The purpose of the current investigation was to examine the relation between cardioballistic impulse and the fluctuations in continuous isometric force production. Subjects produced isometric force via index finger flexion to constant force targets (0.5, 1 and 2 N) with and without visual feedback while beat to beat blood pressure of their middle finger was recorded. Force fluctuations were quantified using distributional statistics. The association between blood pressure oscillations and fluctuations in force output were quantified with coherence analysis. Overall, it was found that force variability (i.e., SD) increased with force level and removal of visual feedback. Coherence values between blood pressure oscillations and force fluctuations were significant and the greatest in the 8-12 Hz bandwidth. There was no effect of force magnitude on the coupling strength between blood pressure oscillations and force production. This coupling was greater in the visual condition. These data suggest that peripheral alterations in blood pressure are related to fluctuations in isometric force production independent of force level and that this interaction is influenced by visual feedback. PMID- 21628727 TI - The reliance on visual feedback for online and offline processing. AB - The objective of the current study was to determine whether the reliance on visual feedback that develops with practice is to due utilizing vision to adjust trajectories during movement execution (i.e., online) and/or to enhance the programming of subsequent trials (i.e., offline). Participants performed a directional aiming task with either vision during the movement, dynamic feedback of the trajectory of the movement or the movement endpoint. The full vision condition was more accurate during practice than the other feedback conditions but suffered a greater decrement in performance when feedback was removed. In addition, the reliance on trajectory feedback was greater compared with the endpoint feedback. It appears that the reliance on visual feedback that develops with practice was due to both online and offline processing. PMID- 21628728 TI - Strength training of one limb increases corticomotor excitability projecting to the contralateral homologous limb. AB - The contralateral transfer of strength following unilateral strength training (ULS) is thought to be due to changes within the nervous system. Using transcranial magnetic stimulation (TMS) we compared corticospinal responses following ULS of the right biceps brachii (BB) projecting to the untrained left BB. Motor evoked potentials (MEPs) were recorded from both BB of 23 individuals pre and post 4 weeks heavy load (80% of 1RM) ULS of right BB. TMS was delivered at intensities below active motor threshold (AMT) to saturation of the MEP (MEPmax). ULS resulted in a 28% increase in 1RM right BB strength, resulting in a 19.2% increase in contralateral strength of the left BB (p = .0001). There was a significant increase in MEP amplitude of 30.3% (p = .03), 33% (p = .05), and 26.5% (p = .01) at AMT, 20% above AMT and MEPmax respectively. No significant differences in silent period were seen at AMT, 20% above AMT or MEPmax. This study has demonstrated increased corticospinal excitability projecting to the untrained arm following heavy load ULS. PMID- 21628729 TI - Evidence for the spotting hypothesis in gymnasts. AB - The goal of this study was to investigate the visual spotting hypothesis in 10 experts and 10 apprentices as they perform back aerial somersaults from a standing position with no preparatory jumps (short flight duration condition) and after some preparatory jumps with a flight time of 1s (long flight duration condition). Differences in gaze behavior and kinematics were expected between experts and apprentices and between experimental conditions. Gaze behavior was measured using a portable and wireless eye-tracking system in combination with a movement-analysis system. Experts exhibited a smaller landing deviation from the middle of the trampoline bed than apprentices. Experts showed higher fixation ratios during the take-off and flight phase. Experts exhibited no blinks in any of the somersaults in both conditions, whereas apprentices showed significant blink ratios in both experimental conditions. The findings suggest that gymnasts can use visual spotting during the back aerial somersault, even when the time of flight is delimited. We conclude that knowledge about gaze-movement relationships may help coaches develop specific training programs in the learning process of the back aerial somersault. PMID- 21628730 TI - Sensorimotor integration for functional recovery and the Bobath approach. AB - Bobath therapy is used to treat patients with neurological disorders. Bobath practitioners use hands-on approaches to elicit and reestablish typical movement patterns through therapist-controlled sensorimotor experiences within the context of task accomplishment. One aspect of Bobath practice, the recovery of sensorimotor function, is reviewed within the framework of current motor control theories. We focus on the role of sensory information in movement production, the relationship between posture and movement and concepts related to motor recovery and compensation with respect to this therapeutic approach. We suggest that a major barrier to the evaluation of the therapeutic effectiveness of the Bobath concept is the lack of a unified framework for both experimental identification and treatment of neurological motor deficits. More conclusive analysis of therapeutic effectiveness requires the development of specific outcomes that measure movement quality. PMID- 21628731 TI - Sensory information utilization and time delays characterize motor developmental pathology in infant sitting postural control. AB - Sitting is one of the first developmental milestones that an infant achieves. Thus measurements of sitting posture present an opportunity to assess sensorimotor development at a young age. Sitting postural sway data were collected using a force plate, and the data were used to train a neural network controller of a model of sitting posture. The trained networks were then probed for sensitivity to position, velocity, and acceleration information at various time delays. Infants with typical development developed a higher reliance on velocity information in control in the anterior-posterior axis, and used more types of information in control in the medial-lateral axis. Infants with delayed development, where the developmental delay was due to cerebral palsy for most of the infants in the study, did not develop this reliance on velocity information, and had less reliance on short latency control mechanisms compared with infants with typical development. PMID- 21628732 TI - [Pathophysiology and treatment of branch atheromatous disease]. PMID- 21628733 TI - [Surgical approach for the tuberculum sellae meningioma: high and low?]. PMID- 21628734 TI - [Visualization of the distal edge of carotid plaque using indocyanine green videoangiography during carotid endarterectomy and the pitfall involved]. AB - OBJECTIVE: Microscope-integrated near-infrared indocyanine green video angiography (ICG-VA) is a new method of intraoperative blood flow assessment. The objective of this study was to evaluate the efficacy of ICG-VA in carotid endarterectomy to visualize the carotid plaque area, especially the distal end. METHODS: Twenty patients who underwent carotid endarterectomy were included. Before arteriotomy and after vessel closure, ICG was injected intravenously. The plaque area and patency of the carotid artery were assessed. RESULTS: In all cases, the distal end of the carotid plaque was identified and the patency of the internal carotid artery was confirmed after excision of plaque. In seventeen cases (85.0%), the proximal end was also identified. The internal surface of carotid plaque was not seen. Some images show reflux of contrast medium into the internal jugular after a left-arm injection. CONCLUSIONS: ICG-VA provides real time information on the carotid plaque area and the patency of arterial vessel during surgical procedure. It is necessary to pay special attention to injection routes, existence of any foreign body and the influence of hemostasis in order to get better contrast. ICD-VA would be a useful adjunct to improve the quality of CEA. PMID- 21628735 TI - [Antitumor effects of gefitinib for metastatic brain tumors from lung carcinomas with EGFR mutation]. AB - OBJECTIVE: Patients with lung cancer harboring epidermal growth factor receptor (EGFR) mutation respond remarkably well to tyrosine kinase inhibitors of the EGFR (EGFR-TKI). We examined the relation of the EGFR mutation and the efficacy of EGFR-TKI for metastatic brain tumors from lung cancer. MATERIALS AND METHODS: Forty-one patients with brain metastases from lung cancer were treated in our hospital from January 2007 to October 2010. Among them, 9 patients were examined on their EGFR mutation of tumors using the PNA-LNA PCR clamp method, and were treated with gefitinib, given orally at a daily dose of 250 mg. The drug efficacy for brain tumors was evaluated with MRI and CT. RESULTS: Seven patients had EGFR mutation (4 in exon 19, and 3 in exon 21). Five patients showed partial response, 3 remained stable, and one had progressive disease. All 5 patients who showed partial response had EGFR mutation. One patient who had progressive disease had no EGFR mutation. Three patients (case 1, 2 and 6) among 5 patients who showed partial response were well controlled only with gefitinib (without radiation). CONCLUSION: This study suggests that the efficacy of EGFR-TKI for metastatic brain tumors from lung cancer is related to the EGFR mutation of tumor. PMID- 21628736 TI - [A case of multiple dural arteriovenous fistulas treated by multiple modalities]. AB - The authors report a rare case of multiple intracranial dural arteriovenous fistulas (DAVF) at separate sinuses. A 70-year-old man was introduced to our hospital complaining of visual disturbance due to bilateral choked disk, headache, and tinnitus. Initial angiography showed DAVFs involving the superior sagittal sinus and bilateral transverse-sigmoid sinuses, and the occlusion of the right jugular vein. The patient developed progressive impairment of visual activity and had high intracranial pressure (ICP) caused by venous hypertension. No cerebral alteration was seen on magnetic resonance imaging. To decrease the high ICP, surgical sinus isolation of the superior sagittal sinus was performed. After the surgery, transvenous embolization was performed to the right transverse sigmoid sinus DAVF. Headache and tinnitus improved after these treatments, but visual activities rapidly declined and he experienced blindness in just a few months. Gamma knife radiosurgery was performed to the residual DAVFs. We discussed the etiology and treatment of the multiple DAVF, and reviewed past literatures. PMID- 21628737 TI - [Aquired stuttering in addition to a recurrence of developmental stuttering caused by brain tumor in the corpus callosum]. AB - We reported a 66-year-old right-handed man, in whom developmental stuttering had almost been cured previously, who suffered a brain tumor. He developed not only left hemiparesis but also speech dysfluency such as acquired stuttering and aphasia. MRI showed a brain tumor in the body of the corpus callosum. The main feature of his stuttering was repetition of syllables. His stuttering was associated without secondary phenomena such as tongue clicking. His stuttering demonstrated no adaptation benefit and no consistency effect. These results suggest that the stuttering in this patient might have characteristics of both developmental and acquired stuttering, which in turn may suggest that after brain tumor, acquired stuttering occurs in addition to a recurrence of developmental stuttering. PMID- 21628738 TI - [Spontaneous disappearance of intracranial arteriovenous malformation after living-donor liver transplantation: a case report]. AB - In this report, the authors describe a 35-year-old male whose intracranial arteriovenous malformation (AVM) spontaneously disappeared about 2 years after successful living-donor liver transplantation for alcohol-induced liver cirrhosis. Preoperative screening MRI revealed intracranial arteriovenous malformation (AVM) around the midbrain. Cerebral angiography demonstrated that the AVM was fed by the paramedian mesencephalic arteries and was drained via the vein of Galen. He successfully underwent living-donor liver transplantation, and his postoperative course was uneventful. Follow-up MRI and MRA revealed spontaneous disappearance of the AVM 27 months after surgery. The authors discuss precisely the underlying mechanism of this rare phenomenon, based on thorough literature review. PMID- 21628739 TI - [Epidermoid tumor originating from the petrous bone: report of two cases]. AB - In case 1, the tumor was incidentally found in the right petrous bone, middle cranial fossa and cerebello-pontine angle. T1 weighted MRI demonstrated a low intense mass and T2 weighted imaging demonstrated hyperintensity. Through the subtemporal extradural approach, an epidermoid in the middle cranial fossa was partially removed. Postoperative course was uneventfull but an episode of rhinorrhea occurred 15 months later. Bone-window CT scan disclosed air cells of the petrous bone were exposed to the previous surgical cavity. Using the same approach, an epidermoid was totally removed. With the sealing of the tumor cavity with the vascularized muscle flap, the patient became free from rhinorrhea. Case 2 had complainted of sensory impairment in the left trigeminal nerve distribution, atrophy of the left temporal and masseter muscle, and diplopia. T1 weighted imaging of MRI demonstrated a low-intense mass in the left petrous bone, middle cranial fossa, temporal lobe, and cerebello-pontine angle, and T2 weighted imaging demonstrated hyperintensity. The intradural tumor under the temporal lobe was removed at another hospital. As the diplopia deteriorated 5 years later, the patient was re-introduced to our hospital. At first, the tumor in the cerebello pontine angle was removed using the left retromastoid lateral suboccipital approach. Later, the tumor in the petrous bone and middle cranial fossa was removed through the left subtemporal extradural approach. With the sealing of the tumor cavity with the vascularized muscle flap, postoperative cerebrospinal fluid leakage was prevented. The epidermoid tumor in the petrous apex is a congenital and rare disease. The obstruction of the petrous air cell and dural defect using the vascularized flap is most important to prevent postoperative cerebrospinal fluid leakage. PMID- 21628740 TI - [A case of ectopic pituitary adenoma occurring in the sphenoid sinus]. AB - Ectopic pituitary adenomas are relatively rare tumors. We present a case of ectopic pituitary adenoma occurring in the sphenoid sinus. A 63-year-old woman was referred to our hospital complaining of headache. She had no endocrinological abnormalities. Magnetic resonance imaging showed a tumor in the sphenoid sinus, adjacent to the sellar floor and appearing as a low-signal on T1-weighted image and a high signal on T2-weighted image. No connection between the normal pituitary gland and tumor was observed. Using an endonasal-transsphenoidal approach assisted with neuro-endoscopy, we performed total removal of the tumor. No connection between the normal pituitary gland and the tumor was found. Histopathological analysis showed a pituitary adenoma. As demonstrated by our case report, differential diagnosis of a tumor occurring in the sphenoid sinus must include consideration of the existence of an ectopic pituitary adenoma. PMID- 21628741 TI - [A case of serous surface papillary carcinoma of the peritoneum metastatic to the brain]. AB - A case of brain metastasis from peritoneal serous surface papillary carcinoma (SSPC) was reported. This 68-year-old woman was admitted to our department because of decreased consciousness level for the last few days. Her medical past history revealed breast cancer and SSPC of the peritoneum at the age of 64. On admission she was comatose and irritable. Serum and urine examination revealed the syndrome of inappropriate secretion of antidiuretic hormone that was treated with strict restriction of water intake. MRI of the brain demonstrated a nonspecific mass in the subcortical area of the right superior parietal lobule. The mass was successfully removed in en bloc fashion. Pathological diagnosis was SSPC that was compatible with the previous diagnosis obtained from the peritoneum four years previously. Although the patient received whole brain radiation therapy postoperatively, her condition deteriorated rapidly. She died four months after brain surgery. SSPC of the peritoneum is a rare malignant tumor that is defined as a primary tumor histologically indistinguishable from serous carcinoma of the ovary, diffusely involving the peritoneal surface but sparing or only superficially invading the ovaries. Because of the prolongation of survival resulting from advanced chemotherapy for SSPC of the peritoneum, more patients live long enough to develop brain metastasis. Therefore, SSPC of the peritoneum should be kept in mind in the differential diagnosis of a primary site for brain metastasis. PMID- 21628742 TI - [A case of open head injury caused by electric saw]. AB - A rare case of traumatic injury caused by an electric saw is herein described. A 75-year-old male patient was admitted to our hospital as an emergency patient. At the time of the patient's admission, he was fully conscious with no neurological deficits. Computed tomography revealed an intracerebral hematoma in the right frontal lobe, traumatic subarachnoid hemorrhage and pneumocephalus. Emergency surgery was performed at 4 hours 30 minutes after the injury. After a right frontal craniectomy, the intracerebral hematoma was completely evacuated. We applied sufficient debridement. The patient had neither intracranial infection nor vascular injury. We believe that the early operation for the sake of safety was therefore effective for this patient. PMID- 21628743 TI - [Congenital anomalies in the central nervous system (7) Chiari malformation]. PMID- 21628744 TI - [A healthy diet is good for the Earth (not only for the humankind)]. PMID- 21628745 TI - [When the screening programme is reticent]. PMID- 21628746 TI - [Protocol on Water and Health (UNECE and WHO)]. PMID- 21628747 TI - [HIA, health impact assessment: a multidisciplinary procedure to support decision making in public health]. PMID- 21628748 TI - [Toward a comparative effectiveness policy in Italy]. PMID- 21628749 TI - [The presence of arsenic in drinking water: any risk for the exposed population?]. PMID- 21628751 TI - Mortality risk in a cohort of subjects reported by authorities for cannabis possession for personal use. Results of a longitudinal study. AB - OBJECTIVE: to study mortality risk in a cohort of cannabis consumers. DESIGN: longitudinal study on a cohort of 2,511 subjects reported to the Drug Addiction Control Force (DACF) for cannabis possession for personal use between 1990 and 2004 residents in the metropolitan area of Bologna. METHODS: to identify people already treated for drug dependence, the cohort was linked with the clinical archive of the public treatment centers for drug addiction (PTC) in the metropolitan area of Bologna and the results were separated into PTC clients (subjects in treatment at a PTC, all heroin users) and non-PTC clients (subjects not in treatment at a PTC, no knowledge regarding other illicit drugs being used besides cannabis). RESULTS: an excess of mortality was observed for all causes in the cohort compared to the general population, both among PTC clients (SMR 14.61 CI 95% 9.21-23.19) and non-clients (SMR 2.43 CI 95% 1.67-3.55). Among PTC clients the highest and most statistically significant SMRs were for overdose, suicide, and AIDS. Among non PTC clients there was an excess for traffic accidents and overdose. CONCLUSION: the results of the study show an elevated risk of death for consumers of cannabis, a percentage of which probably also consumed other substances, and a very few which presented themselves for treatment at a public drug treatment center. PMID- 21628750 TI - [Skin cancer risk factors in childhood: findings of a survey conducted within Italian areas with a different incidence of melanoma]. AB - OBJECTIVE: to evaluate the association between phenotype and sun sensitivity factors, sun protection behavior, ethnicity and the area of residence in school aged children. DESIGN: a cross-sectional study in the framework of a survey of children using a self-administered questionnaire to be filled in by parents. SETTING AND PARTICIPANTS: 56390 children attending primary schools located in the Italian provinces of Brindisi, Rome, Forli and Genova, in the period between 1998 2002. MAIN OUTCOME MEASURES: Odds Ratios (ORs) and their relative 95% Confidence Intervals (95% CI) are to be computed through univariate and multivariate logistic regression models. RESULTS: "FOTO positive" phenotype, a proxy variable of the fair phenotype, was directly and significantly associated with the tendency to sunburn (OR 4.75; 95% CI 4.54-4.96), the use of sunscreens (OR 1.79; 95% CI 1.63-1.97), the number of grandparents born in the northern areas (OR 1.63; 95% CI 1.45-1.83, for four northern grandparents versus none), the presence of freckles on the face (OR 1.62; 95% CI 1.53-1.72) and of naevi on the left forearm (OR 1.20; 95% CI 1.15-1.26). A positive association was also found for the residence in the northern areas using the area of Brindisi as the reference category, (Rome: OR 1.02; 95% CI 0.95-1.10; Forli: OR 1.05; 95% CI 0.96-1.15; Genova: OR 1.16; 95% CI 1.08-1.26); the ORs increase with latitude, as does the incidence rate of melanoma in Italy. An inverse association was observed with the male sex (OR 0.92; 95% CI 0.88-0.96), the increase of school-class level (OR 0.65; 95% CI 0.60-0.69, for the highest versus the lowest school-class level) and the ability to tan (OR 0.40; 95% CI 0.36-0.43). CONCLUSION: these findings confirmed that the fairness phenotype is associated with other skin cancer risk factors in children and pointed out that the high-risk phenotype has a geographical distribution consistent with the pattern of melanoma incidence in the Italian areas covered by the study. PMID- 21628752 TI - [Contamination of the food products by lead, cadmium and copper in the area at risk of Gela (Sicily)]. AB - OBJECTIVE: to make an initial evaluation on the impact of environmental pollution on the food safety of some agricultural products in the area of Gela, an official area of environmental crisis according to Italian legislation. SETTING: Gela (Caltanisetta ) Sicily MAIN OUTCOME MEASURES: the investigation looked at the contamination of the food chain, one of the main sources of exposure for the local population. A sample of 13 artichokes and 13 tomatoes where sprayed with contaminated water and the levels of Pb (II), Cd (II) end Cu (II) where then measured. RESULTS AND CONCLUSIONS: the analysis of the samples investigated shows a high level of metal contamination which can be linked to both the polluted water and air. It is therefore easier to forecast a scenario of exposure for the local population through the food chain. PMID- 21628753 TI - [Risk factors analysis for elective caesarean section in Campania region (Italy)]. AB - OBJECTIVE: to assess both clinical and non clinical factors associated with elective caesarean section practice and to highlight those typologies of expectant mothers who underwent elective caesarean delivery even in the absence of clinical risk factors. SETTING AND PARTICIPANTS: Birth Certificates archive has been used as data source: 62888 available records of all the births that occurred in the Campania region in 2005 both in public and private hospitals. Only those births pertained to women without previous caesarean sections were analyzed. MAIN OUTCOME MEASURES: type of delivery, dichotomized as elective caesarean section and vaginal delivery. RESULTS: after adjusting the data for known risk factors, elective caesarean section practice emerged as much more prevalent in private hospitals than in public ones (53.1% vs 24.3%, p<0.001). A multivariate logistic regression model showed a significant association between primiparity and elective caesarean section. This association was stronger in private hospitals (OR for interaction between parity and hospital typology 1.81, CI 95% 1.62-2.04). Being older than 34 years, having recoursed to medically assisted procreation and a previous voluntary termination of pregnancy cause an increase in the odd of resorting to elective caesarean section (p<0.01). Being foreigner, high educational qualification and young age were protective factors for elective caesarean section (respectively, OR 0.36 CI 95% 0.31-0.42, OR 0.81 CI 95% 0.77-0.86 e OR 0.84 CI 95% 0.78 -0.90). Five different typologies of primiparous women were found by means of a hierarchical classification procedure. A high percentage of elective caesarean section was found in women assisted in private structure with no clinical risk. CONCLUSIONS: in Campania there is an excess of elective caesarean sections among primiparous women without manifest clinical risk factors; this excess is higher when the delivery occurs in Private Hospitals. Public health policy and actions aimed at reducing caesarean section rates should be addressed mainly to this typology of women. PMID- 21628754 TI - [Obesity among the adult population in Italy]. AB - OBJECTIVE: analysis of the prevalence and factors of obesity among the adult population in Italy. MATERIALS: microdata from the Multipurpose Survey on Households "Aspects of daily life 2007", carried out by Istat on a sample of nearly 19 thousands households. METHODS: multiple logistic regression. RESULTS: the risk of obesity is associated to individuals sociodemographic characteristics. Physical and cultural activity are negatively associated with obesity. The stratified analyses highlight differences by gender and age in the factors associated to higher risks of obesity. CONCLUSIONS: the prevention and treatment of obesity require the cooperation of several specialists, not exclusively from the health-sector. PMID- 21628755 TI - [Test and programme sensitivities of screening for colorectal cancer in Reggio Emilia]. AB - OBJECTIVE: to estimate the sensitivity of the immunochemical test for faecal occult blood (FOBT) and the sensitivity of the colorectal tumour screening programme in the province of Reggio Emilia. DESIGN AND SETTINGS: retrospective cohort study, including a sample of 80,357 people of both genders, aged 50-69, who underwent FOBT, during the first round of the screening programme in the province of Reggio Emilia, from April 2005 to December 2007. MAIN OUTCOME MEASURES: incidence of interval cancer. The proportional incidence method was used to estimate the sensitivity of FOBT and of the screening programme. Data were stratified according to gender, age and year of interval. RESULTS AND CONCLUSIONS: the overall sensitivity of FOBT was 73.2% (95%IC 63.8-80.7). The sensitivity of FOBT was lower in females (70.5% vs 75.1%), higher in the 50-59 age group (78.6% vs 70.2%) and higher in the colon than rectum (75.1% vs 68.9%). The test had a significantly higher sensitivity in the 1st year of interval than in the 2nd (84.4% vs 60.5%; RR=0.39, 95%IC 0.22-0.70), a difference which was confirmed, also when data were stratified according to gender. The overall sensitivity of the programme is 70.9% (95%IC 61.5-78.5). No statistically significant differences were shown, if data were stratified according to gender, age or site. Again the sensitivity in the 1st year was significantly higher than in the 2nd year of interval (83.2% vs 57.0%; RR=0.41, 95%IC 0.24-0.69). Overall our data confirmed the findings of similar Italian studies, despite subgroup analysis showed some differences in sensitivity in our study. PMID- 21628756 TI - [Geographical and temporal trends in infant mortality in Italy and current limits of the routine data]. AB - OBJECTIVE: to compare trends in infant, neonatal and post-neonatal mortality in Italy from 1991 to 2005 both at the national level and among the three Italian large geographical macro-areas (North, Center, South-Islands). DESIGN: observational study based on routine data. SETTING AND PARTICIPANTS: the analysis was performed on the cohort of newborns (up to the first age of life) in Italy from 1991 to 2005. MAIN OUTCOME MEASURES: infant, neonatal and post-neonatal mortality rate calculated for 3-year and 5-year periods, relative risks, attributable fraction. RESULTS: during the study period infant mortality rates significantly decreased from 7.72 to 3.91 per 1,000 births, neonatal mortality rates from 5.87 to 2.84 per 1,000 births, and postneonatal mortality rates from 1.85 to 1.08. Despite these significant reductions, important disparities persist in different geographical areas within Italy. In particular, rates appears to be much higher in the southern regions of the country: during the period 2001-2005 the excess of mortality in the South comparing with the North was 37%. Since 1998, following a change in legislation, individual matching of Certificates of Delivery Care (CedAP) and Death Certificates during the first year of life, at a national level, is not possible. CONCLUSION: during the period 1991-2005 Italy experienced significant infant mortality reduction, but important geographical disparities still remain. In order to investigate these disparities and the determinants of infant mortality in Italy, the lack of routine data could represent an important limit to conduct update epidemiologic studies. PMID- 21628757 TI - [The role of Health Impact Assessment (HIA) in the decision-making]. AB - The review describes the evolution of the environmental policies in Italy underlying current delays and discords between the environment and its effects on human health. It also tries to define the major barriers to the introduction of the health impact assessment (HIA) in government policy-making process in Italy; the lack of regulations and the lack of methods to perform an HIA are the main reasons for its limited use. Epidemiology is a fundamental tool to produce effective health impact assessments along with a deep knowledge of medicine, environmental hygiene and risk communication. In conclusion, should the Italian scientific community agreed on a common and constructive approach avoiding possible exploitations and conflicts of interest, HIA could be a valuable method for helping to develop environmental policies and assisting policy-makers in important decisions. PMID- 21628758 TI - [Moniter Project: a model of HIA for Incinerators]. AB - The Emilia-Romagna regional government decided to develop a monitoring and surveillance system, known as MonITER (Incineration Monitoring on the Emilia Romagna Territory), in 8 areas characterized by the presence of municipal waste incinerating plants.The project was divided into 7 lines, among these the number 6 developed a procedure of Health Impact Assessments (HIA) to support the planning of new facilities for incineration or combustion. Line number 6 is divided into 3 actions n action 1 aimed to developing and validating a participative HIA methodology, as well tools for the impact assessment and recommendations for implementing HIA on future plants; n action 2 analysed connected issues on communication; n action 3 was focused to developing a methodology for integrating HIA models into existing tools for mandatory impact assessment.Tailoring the HIA model development to the waste management context has highlighted the fundamental differences of the HIA approach with the other mandatory assessment tools, being theoretically founded on the principles of public involvement, equity, sustainable development and ethical use of evidence. PMID- 21628759 TI - [Problems due to the prolonged permanence of asbestos-containing materials]. AB - In 1992 a law banned excavation, importation, production and trade of new asbestos-containing materials. Nonetheless the use of already made asbestos containing materials hadn't been banned. Now it's time to think about the possible health impact on the Italian population, and workers in particular, for the prolonged permanence of asbestos-containing materials, in civil and industrial areas. PMID- 21628760 TI - [Discharge data for surveillance of cancer care]. AB - The debate on the use of hospital discharge records in epidemiological studies focused in Italy on biased estimates of tumor incidence and survival produced through analyses of hospitalization data. By contrast, despite limitations in completeness and accuracy which may increase with the shift of delivered health care towards non-hospital settings, hospital discharge records are among the most useful - and still underused - tools for the evaluation of patterns of care in cancer patients. Such analyses are rarely carried out in Italy; a striking demonstration comes from cancer surgery which, with the exception of breast cancer surgery, has been the issue of few population-based studies. Experiences from the United States show how analyses of hospital discharge records, cancer registries data, their linked archives, as well as of hospital-based registries, can be effective and complementary tools for the surveillance of major patterns in cancer therapy. PMID- 21628761 TI - [Multiple testing and subgroup analysis (what's wrong in always searching for significant results)]. PMID- 21628762 TI - [Water and climate change]. PMID- 21628763 TI - [Respect of Smoking ban in public places is increasing, but relevant differences among Northern, Central and Southern Italy still exist]. PMID- 21628764 TI - [1991-2005: surveillance increases and thyroid cancer incidence doubles]. PMID- 21628765 TI - [Risk variability and rate variability]. PMID- 21628766 TI - [Comparative evaluation of health services outcomes]. AB - This introductory guide represents an operative tool to conduct epidemiological studies in the area of comparative outcomes evaluation. It is based on the experience of epidemiological research in this field conducted in Italy within national (BPAC-Esiti del bypass aortocoronarico, Progetto mattoni outcome, Progetto Progressi) or regional (P.Re.Val.E. Programma Regionale di Valutazione degli Esiti, Lazio) health care outcomes projects and the National outcome programme. This guide is aimed to all those interested in conducting or interpreting health care outcomes studies within different levels of the Italian NHS. It gives an introductory description of the operative steps to build outcome indicators and to perform comparative analyses, with the general objective of measuring and promoting improvement in health care. A specific emphasis is given to the use of routinely collected health care databases that have found widespread use for epidemiological purposes. This guide has two parts: part A includes an introduction and comments on critical methodological points, part B shows three example of epidemiological studies (A. Complications after cholecystectomy: comparison between two surgical techniques, B. 30-day mortality after acute myocardial infarction: comparison among hospitals, C. 30-day mortality after acute myocardial infarction: comparison between time periods). The online version of this guide is organised as a hypertext as practical instrument of appraisal. PMID- 21628767 TI - The feasibility of polychromatic cone-beam x-ray fluorescence computed tomography (XFCT) imaging of gold nanoparticle-loaded objects: a Monte Carlo study. AB - A recent study investigated the feasibility to develop a bench-top x-ray fluorescence computed tomography (XFCT) system capable of determining the spatial distribution and concentration of gold nanoparticles (GNPs) in vivo using a diagnostic energy range polychromatic (i.e. 110 kVp) pencil-beam source. In this follow-up study, we examined the feasibility of a polychromatic cone-beam implementation of XFCT by Monte Carlo (MC) simulations using the MCNP5 code. In the current MC model, cylindrical columns with various sizes (5-10 mm in diameter) containing water loaded with GNPs (0.1-2% gold by weight) were inserted into a 5 cm diameter cylindrical polymethyl methacrylate phantom. The phantom was then irradiated by a lead-filtered 110 kVp x-ray source, and the resulting gold fluorescence and Compton-scattered photons were collected by a series of energy sensitive tallies after passing through lead parallel-hole collimators. A maximum likelihood iterative reconstruction algorithm was implemented to reconstruct the image of GNP-loaded objects within the phantom. The effects of attenuation of both the primary beam through the phantom and the gold fluorescence photons en route to the detector were corrected during the image reconstruction. Accurate images of the GNP-containing phantom were successfully reconstructed for three different phantom configurations, with both spatial distribution and relative concentration of GNPs well identified. The pixel intensity of regions containing GNPs was linearly proportional to the gold concentration. The current MC study strongly suggests the possibility of developing a bench-top, polychromatic, cone beam XFCT system for in vivo imaging. PMID- 21628769 TI - Neural cell-cell and cell-substrate adhesion through N-cadherin, N-CAM and L1. AB - In this study neural (N)-cadherin, neural cell adhesion molecule (N-CAM) and L1 proteins and their antibody equivalents were covalently immobilized on a polyethylene-imine (PEI)-coated glass surface to form neuron-adhesive coatings. Impedance sensing and (supplementary) image analysis were used to monitor the effects of these CAMs. Immobilization of high concentrations of both N-cadherin protein and antibody led to good adhesion of neurons to the modified surface, better than surfaces treated with 30.0 and 100.0 ug ml(-1) N-CAM protein and antibody. L1 antibody and protein coating revealed no significant effect on neuronal cell-substrate adhesion. In a second series of combinatorial experiments, we used the same antibodies and proteins as medium-additives to inhibit cell-cell adhesion between neurons. Adhesion of neurons cultured on N cadherin protein or antibody-modified surfaces was lowered by the addition of a soluble N-cadherin protein and antibody to the culturing medium, accelerating neuronal aggregation. The presence of a soluble N-CAM antibody or protein had no effect on the adhesion of neuronal cells on a N-cadherin protein-modified surface. On a N-cadherin antibody-coated surface, the addition of a soluble N-CAM protein led to cell death of neurons after 48 h, while a N-CAM antibody had no effect. In the presence of a soluble N-cadherin protein and antibody the aggregation of neurons was inhibited, both on N-CAM protein and N-CAM antibody modified surfaces. Neurons cultured on immobilized antibodies were less affected by the addition of soluble CAM blockers than neurons cultured on immobilized proteins, indicating that antibody-protein bonds are more stable compared to protein-protein bonds. PMID- 21628768 TI - Calcium channel dynamics limit synaptic release in response to prosthetic stimulation with sinusoidal waveforms. AB - Extracellular electric stimulation with sinusoidal waveforms has been shown to allow preferential activation of individual types of retinal neurons by varying stimulus frequency. It is important to understand the mechanisms underlying this frequency dependence as a step toward improving methods of preferential activation. In order to elucidate these mechanisms, we implemented a morphologically realistic model of a retinal bipolar cell and measured the response to extracellular stimulation with sinusoidal waveforms. We compared the frequency response of a passive membrane model to the kinetics of voltage-gated calcium channels that mediate synaptic release. The passive electrical properties of the membrane exhibited lowpass filtering with a relatively high cutoff frequency (nominal value = 717 Hz). This cutoff frequency was dependent on intra axonal resistance, with shorter and wider axons yielding higher cutoff frequencies. However, we found that the cutoff frequency of bipolar cell synaptic release was primarily limited by the relatively slow opening kinetics of L- and T type calcium channels. The cutoff frequency of calcium currents depended nonlinearly on stimulus amplitude, but remained lower than the cutoff frequency of the passive membrane model for a large range of membrane potential fluctuations. These results suggest that while it may be possible to modulate the membrane potential of bipolar cells over a wide range of stimulus frequencies, synaptic release will only be initiated at the lower end of this range. PMID- 21628770 TI - Controlling selective stimulations below a spinal cord hemisection using brain recordings with a neural interface system approach. AB - In this work we address the use of realtime cortical recordings for the generation of coherent, reliable and robust motor activity in spinal-lesioned animals through selective intraspinal microstimulation (ISMS). The spinal cord of adult rats was hemisectioned and groups of multielectrodes were implanted in both the central nervous system (CNS) and the spinal cord below the lesion level to establish a neural system interface (NSI). To test the reliability of this new NSI connection, highly repeatable neural responses recorded from the CNS were used as a pattern generator of an open-loop control strategy for selective ISMS of the spinal motoneurons. Our experimental procedure avoided the spontaneous non controlled and non-repeatable neural activity that could have generated spurious ISMS and the consequent undesired muscle contractions. Combinations of complex CNS patterns generated precisely coordinated, reliable and robust motor actions. PMID- 21628771 TI - Automatic subthalamic nucleus detection from microelectrode recordings based on noise level and neuronal activity. AB - Microelectrode recording (MER) along surgical trajectories is commonly applied for refinement of the target location during deep brain stimulation (DBS) surgery. In this study, we utilize automatically detected MER features in order to locate the subthalamic nucleus (STN) employing an unsupervised algorithm. The automated algorithm makes use of background noise level, compound firing rate and power spectral density along the trajectory and applies a threshold-based method to detect the dorsal and the ventral borders of the STN. Depending on the combination of measures used for detection of the borders, the algorithm allocates confidence levels for the annotation made (i.e. high, medium and low). The algorithm has been applied to 258 trajectories obtained from 84 STN DBS implantations. MERs used in this study have not been pre-selected or pre processed and include all the viable measurements made. Out of 258 trajectories, 239 trajectories were annotated by the surgical team as containing the STN versus 238 trajectories by the automated algorithm. The agreement level between the automatic annotations and the surgical annotations is 88%. Taking the surgical annotations as the golden standard, across all trajectories, the algorithm made true positive annotations in 231 trajectories, true negative annotations in 12 trajectories, false positive annotations in 7 trajectories and false negative annotations in 8 trajectories. We conclude that our algorithm is accurate and reliable in automatically identifying the STN and locating the dorsal and ventral borders of the nucleus, and in a near future could be implemented for on-line intra-operative use. PMID- 21628773 TI - Uncertainties in planned dose due to the limited voxel size of the planning CT when treating lung tumors with proton therapy. AB - Dose calculation for lung tumors can be challenging due to the low density and the fine structure of the geometry. The latter is not fully considered in the CT image resolution used in treatment planning causing the prediction of a more homogeneous tissue distribution. In proton therapy, this could result in predicting an unrealistically sharp distal dose falloff, i.e. an underestimation of the distal dose falloff degradation. The goal of this work was the quantification of such effects. Two computational phantoms resembling a two dimensional heterogeneous random lung geometry and a swine lung were considered applying a variety of voxel sizes for dose calculation. Monte Carlo simulations were used to compare the dose distributions predicted with the voxel size typically used for the treatment planning procedure with those expected to be delivered using the finest resolution. The results show, for example, distal falloff position differences of up to 4 mm between planned and expected dose at the 90% level for the heterogeneous random lung (assuming treatment plan on a 2 * 2 * 2.5 mm(3) grid). For the swine lung, differences of up to 38 mm were seen when airways are present in the beam path when the treatment plan was done on a 0.8 * 0.8 * 2.4 mm(3) grid. The two-dimensional heterogeneous random lung phantom apparently does not describe the impact of the geometry adequately because of the lack of heterogeneities in the axial direction. The differences observed in the swine lung between planned and expected dose are presumably due to the poor axial resolution of the CT images used in clinical routine. In conclusion, when assigning margins for treatment planning for lung cancer, proton range uncertainties due to the heterogeneous lung geometry and CT image resolution need to be considered. PMID- 21628772 TI - Simultaneous recording of rat auditory cortex and thalamus via a titanium-based, microfabricated, microelectrode device. AB - Direct recording from sequential processing stations within the brain has provided opportunity for enhancing understanding of important neural circuits, such as the corticothalamic loops underlying auditory, visual, and somatosensory processing. However, the common reliance upon microwire-based electrodes to perform such recordings often necessitates complex surgeries and increases trauma to neural tissues. This paper reports the development of titanium-based, microfabricated, microelectrode devices designed to address these limitations by allowing acute recording from the thalamic nuclei and associated cortical sites simultaneously in a minimally invasive manner. In particular, devices were designed to simultaneously probe rat auditory cortex and auditory thalamus, with the intent of recording auditory response latencies and isolated action potentials within the separate anatomical sites. Details regarding the design, fabrication, and characterization of these devices are presented, as are preliminary results from acute in vivo recording. PMID- 21628774 TI - 3D current source density imaging based on the acoustoelectric effect: a simulation study using unipolar pulses. AB - It is of importance to image electrical activity and properties of biological tissues. Recently hybrid imaging modality combing ultrasound scanning and source imaging through the acoustoelectric (AE) effect has generated considerable interest. Such modality has the potential to provide high spatial resolution current density imaging by utilizing the pressure-induced AE resistivity change confined at the ultrasound focus. In this study, we investigate a novel three dimensional (3D) ultrasound current source density imaging approach using unipolar ultrasound pulses. Utilizing specially designed unipolar ultrasound pulses and by combining AE signals associated to the local resistivity changes at the focusing point, we are able to reconstruct the 3D current density distribution with the boundary voltage measurements obtained while performing a 3D ultrasound scan. We have shown in computer simulation that using the present method it is feasible to image with high spatial resolution an arbitrary 3D current density distribution in an inhomogeneous conductive media. PMID- 21628775 TI - MRI-guided tumor tracking in lung cancer radiotherapy. AB - Precise tracking of lung tumor motion during treatment delivery still represents a challenge in radiation therapy. Prototypes of MRI-linac hybrid systems are being created which have the potential of ionization-free real-time imaging of the tumor. This study evaluates the performance of lung tumor tracking algorithms in cine-MRI sagittal images from five healthy volunteers. Visible vascular structures were used as targets. Volunteers performed several series of regular and irregular breathing. Two tracking algorithms were implemented and evaluated: a template matching (TM) algorithm in combination with surrogate tracking using the diaphragm (surrogate was used when the maximum correlation between the template and the image in the search window was less than specified), and an artificial neural network (ANN) model based on the principal components of a region of interest that encompasses the target motion. The mean tracking error e and the error at 95% confidence level e(95) were evaluated for each model. The ANN model led to e = 1.5 mm and e(95) = 4.2 mm, while TM led to e = 0.6 mm and e(95) = 1.0 mm. An extra series was considered separately to evaluate the benefit of using surrogate tracking in combination with TM when target out-of-plane motion occurs. For this series, the mean error was 7.2 mm using only TM and 1.7 mm when the surrogate was used in combination with TM. Results show that, as opposed to tracking with other imaging modalities, ANN does not perform well in MR-guided tracking. TM, however, leads to highly accurate tracking. Out-of-plane motion could be addressed by surrogate tracking using the diaphragm, which can be easily identified in the images. PMID- 21628776 TI - Standing adult human phantoms based on 10th, 50th and 90th mass and height percentiles of male and female Caucasian populations. AB - Computational anthropomorphic human phantoms are useful tools developed for the calculation of absorbed or equivalent dose to radiosensitive organs and tissues of the human body. The problem is, however, that, strictly speaking, the results can be applied only to a person who has the same anatomy as the phantom, while for a person with different body mass and/or standing height the data could be wrong. In order to improve this situation for many areas in radiological protection, this study developed 18 anthropometric standing adult human phantoms, nine models per gender, as a function of the 10th, 50th and 90th mass and height percentiles of Caucasian populations. The anthropometric target parameters for body mass, standing height and other body measures were extracted from PeopleSize, a well-known software package used in the area of ergonomics. The phantoms were developed based on the assumption of a constant body-mass index for a given mass percentile and for different heights. For a given height, increase or decrease of body mass was considered to reflect mainly the change of subcutaneous adipose tissue mass, i.e. that organ masses were not changed. Organ mass scaling as a function of height was based on information extracted from autopsy data. The methods used here were compared with those used in other studies, anatomically as well as dosimetrically. For external exposure, the results show that equivalent dose decreases with increasing body mass for organs and tissues located below the subcutaneous adipose tissue layer, such as liver, colon, stomach, etc, while for organs located at the surface, such as breasts, testes and skin, the equivalent dose increases or remains constant with increasing body mass due to weak attenuation and more scatter radiation caused by the increasing adipose tissue mass. Changes of standing height have little influence on the equivalent dose to organs and tissues from external exposure. Specific absorbed fractions (SAFs) have also been calculated with the 18 anthropometric phantoms. The results show that SAFs decrease with increasing height and increase with increasing body mass. The calculated data suggest that changes of the body mass may have a significant effect on equivalent doses, primarily for external exposure to organs and tissue located below the adipose tissue layer, while for superficial organs, for changes of height and for internal exposures the effects on equivalent dose are small to moderate. PMID- 21628777 TI - Quantifying lung shunting during planning for radio-embolization. AB - A method is proposed for accurate quantification of lung uptake during shunt studies for liver cancer patients undergoing radio-embolization. The current standard for analysis of [(99m)Tc]-MAA shunt studies is subjective and highly variable. The technique proposed in this work involves a small additional peripheral intravenous injection of macroaggregated albumin (MAA) and two additional static acquisitions (before and after injection) to quantify the absolute activity in the lungs as a result of arterio-venous shunting. Such quantification also allows for estimates of absorbed dose to lung tissue at the time of treatment based on MIRD formalism. The method was used on six radio embolization patients attending the department for lung shunt analysis. Quantitative values for each were compared to a previously validated technique using fully quantitative SPECT/CT imaging, treated as the gold standard. The average difference between absolute activity shunted to the lungs calculated by the proposed technique compared to the previously validated technique was found to be 2%, with a range of (1-8)%. The proposed method is simple and fast, allowing for accurate quantification of lung shunting and estimates of absorbed dose to lung tissue at treatment, and may one day be used in a one-stop procedure for planning and therapy in a single interventional procedure. PMID- 21628778 TI - GPU-based iterative cone-beam CT reconstruction using tight frame regularization. AB - The x-ray imaging dose from serial cone-beam computed tomography (CBCT) scans raises a clinical concern in most image-guided radiation therapy procedures. It is the goal of this paper to develop a fast graphic processing unit (GPU)-based algorithm to reconstruct high-quality CBCT images from undersampled and noisy projection data so as to lower the imaging dose. For this purpose, we have developed an iterative tight-frame (TF)-based CBCT reconstruction algorithm. A condition that a real CBCT image has a sparse representation under a TF basis is imposed in the iteration process as regularization to the solution. To speed up the computation, a multi-grid method is employed. Our GPU implementation has achieved high computational efficiency and a CBCT image of resolution 512 * 512 * 70 can be reconstructed in ~5 min. We have tested our algorithm on a digital NCAT phantom and a physical Catphan phantom. It is found that our TF-based algorithm is able to reconstruct CBCT in the context of undersampling and low mAs levels. We have also quantitatively analyzed the reconstructed CBCT image quality in terms of the modulation-transfer function and contrast-to-noise ratio under various scanning conditions. The results confirm the high CBCT image quality obtained from our TF algorithm. Moreover, our algorithm has also been validated in a real clinical context using a head-and-neck patient case. Comparisons of the developed TF algorithm and the current state-of-the-art TV algorithm have also been made in various cases studied in terms of reconstructed image quality and computation efficiency. PMID- 21628779 TI - A 4 MV flattening filter-free beam: commissioning and application to conformal therapy and volumetric modulated arc therapy. AB - Recent studies have indicated that radiotherapy treatments undertaken on a flattening filter-free (FFF) linear accelerator have a number of advantages over treatments undertaken on a conventional linear accelerator. In addition, 4 MV photon beams may give improved isodose coverage for some treatment volumes at air/tissue interfaces, compared to when utilizing the clinical standard of 6 MV photons. In order to investigate these benefits, FFF beams were established on an Elekta Beam Modulator linear accelerator for 4 MV photons. Commissioning beam data were obtained for open and wedged fields. The measured data were then imported into a treatment planning system and a beam model was commissioned. The beam model was optimized to improve dose calculations at shallow, clinically relevant depths. Following verification, the beam model was utilized in a treatment planning study, including volumetric modulated arc therapy, for a selection of lung, breast/chest wall and larynx patients. Increased dose rates of around 800 MU min(-1) were recorded for open fields (relative to 320 MU min(-1) for filtered open fields) and reduced head scatter was inferred from output factor measurements. Good agreement between planned and delivered dose was observed in verification of treatment plans. The planning study indicated that with a FFF beam, equivalent (and in some cases improved) isodose profiles could be achieved for small lung and larynx treatment volumes relative to 4 MV filtered treatments. Furthermore, FFF treatments with wedges could be replicated using open fields together with an 'effective wedge' technique and isocentre shift. Clinical feasibility of a FFF beam was therefore demonstrated, with beam modelling, treatment planning and verification being successfully accomplished. PMID- 21628782 TI - Helium-vacancy cluster in a single bcc iron crystal lattice. AB - The properties of the cluster He(n)V, an iron vacancy with an increasing number of He atoms, is studied with molecular statics and molecular dynamics simulations. A study of the binding energy of the self-interstitial atom (SIA) and the He, shows that from n = 6 the He(n)V cluster is stable and cannot shrink anymore, and from n = 16 the He(n)V(2) cluster is stabilized by the emission of SIA in the form of a (110) dumbbell. Calculation of the pressure exercised by the He(n)V cluster shows local peak normal stress and shear stress values up to 9 GPa and 4 GPa, respectively. The local configurations of He(n)V suggest that with increasing helium content, a high symmetry configuration close to a face centered cubic lattice is formed. PMID- 21628780 TI - Effects of pulmonary inhalation on hyperpolarized krypton-83 magnetic resonance T1 relaxation. AB - The (83)Kr magnetic resonance (MR) relaxation time T(1) of krypton gas in contact with model surfaces was previously found to be highly sensitive to surface composition, surface-to-volume ratio, and surface temperature. The work presented here explored aspects of pulmonary (83)Kr T(1) relaxation measurements in excised lungs from healthy rats using hyperpolarized (hp) (83)Kr with approximately 4.4% spin polarization. MR spectroscopy without spatial resolution was applied to the ex vivo lungs that actively inhale hp (83)Kr through a custom designed ventilation system. Various inhalation schemes were devised to study the influence of anatomical dead space upon the measured (83)Kr T(1) relaxation times. The longitudinal (83)Kr relaxation times in the distal airways and the respiratory zones were independent of the lung inhalation volume, with T(1) = 1.3 s and T(1) = 1.0 s, depending only on the applied inhalation scheme. The obtained data were highly reproducible between different specimens. Further, the (83)Kr T(1) relaxation times in excised lungs were unaffected by the presence of up to 40% oxygen in the hp gas mixture. The results support the possible importance of (83)Kr as a biomarker for evaluating lung function. PMID- 21628783 TI - Residual resistance of 2D and 3D structures and Joule heat release. AB - We consider a residual resistance and Joule heat release in 2D nanostructures as well as in ordinary 3D conductors. We assume that elastic scattering of conduction electrons by lattice defects is predominant. Within a rather intricate situation in such systems we discuss in detail two cases. (1) The elastic scattering alone (i.e. without regard of inelastic mechanisms of scattering) leads to a transition of the mechanical energy (stored by the electrons under the action of an electric field) into heat in a traditional way. This process can be described by the Boltzmann equation where it is possible to do the configuration averaging over defect positions in the electron-impurity collision term. The corresponding conditions are usually met in metals. (2) The elastic scattering can be considered with the help of the standard electron-impurity collision integral only in combination with some additional averaging procedure (possibly including inelastic scattering or some mechanisms of electron wavefunction phase destruction). This situation is typical for degenerate semiconductors with a high concentration of dopants and conduction electrons. Quite often, heat release can be observed via transfer of heat to the lattice, i.e. via inelastic processes of electron-phonon collisions and can take place at distances much larger than the size of the device. However, a direct heating of the electron system can be registered too by, for instance, local measurements of the current noise or direct measurement of an electron distribution function. PMID- 21628784 TI - Observation of a level crossing in a molecular nanomagnet using implanted muons. AB - We have observed an electronic energy level crossing in a molecular nanomagnet (MNM) using muon spin relaxation. This effect, not observed previously despite several muon studies of MNM systems, provides further evidence that the spin relaxation of the implanted muon is sensitive to the dynamics of the electronic spin. Our measurements on a broken ring MNM [H(2)N(t)Bu(is)Pr][Cr(8)CdF(9)(O(2)CC(CH(3))(3))(18)], which contains eight Cr ions, show clear evidence for the S = 0 --> S = 1 transition that takes place at B(c) = 2.3 T. The crossing is observed as a resonance-like dip in the average positron asymmetry and also in the muon spin relaxation rate, which shows a sharp increase in magnitude at the transition and a peak centred within the S = 1 regime. PMID- 21628785 TI - Magnetic field barriers in graphene: an analytically solvable model. AB - We study the dynamics of carriers in graphene subjected to an inhomogeneous magnetic field. For a magnetic field with a hyperbolic profile the corresponding Dirac equation can be analyzed within the formalism of supersymmetric quantum mechanics, and leads to an exactly solvable model. We study in detail the bound state spectrum. For a narrow barrier the spectrum is characterized by a few bands, except for the zero energy level that remains degenerated. As the width of the barrier increases we can track the band's evolution into the degenerated Landau levels. In the scattering regime a simple analytical formula is obtained for the transmission coefficient, this result allows us to identify the resonant conditions at which the barrier becomes transparent. PMID- 21628786 TI - Detailed analysis of water structure in a solvent mediated electron tunneling mechanism. AB - This work aims at describing the water structure characteristics that influence the electron transfer superexchange mechanism by explicitly calculating the solvent mediated conductance between the donor and acceptor in a generic pair. The method employed here is based on the non-equilibrium Green function formalism for calculating the conductance over solvent trajectories previously determined by molecular dynamics methods. A non-exponential dependence of the conductance is observed with respect to the distance between the donor and the acceptor. Local fluctuations of the solvent structure are responsible for the non-monotonic dependence, mainly due to the formation of solvent bridges that act as a molecular wire connecting the sites. This shortcutting phenomenon is observed for certain ranges of distances between the donor and acceptor in the pair. Charge on the sites strongly affects the local solvent structure and causes qualitative changes in the distance dependence of the tunneling probability. PMID- 21628787 TI - Synthesis of cubic SrCoO3 single crystal and its anisotropic magnetic and transport properties. AB - A large-size single crystal of nearly stoichiometric SrCoO(3) was prepared with a two-step method combining the floating-zone technique and subsequent high oxygen pressure treatment. SrCoO(3) crystallizes in a cubic perovskite structure with space group Pm3m, and displays an itinerant ferromagnetic behavior with the Curie temperature of 305 K. The easy magnetization axis is found to be along the [111] direction, and the saturation moment is 2.5 u(B)/f.u., in accord with the picture of the intermediate spin state. The resistivity at low temperatures (T) is proportional to T(2), indicative of the possible effect of orbital fluctuation in the intermediate spin ferromagnetic metallic state. Unusual anisotropic magnetoresistance is also observed and its possible origin is discussed. PMID- 21628788 TI - Influence of B-site disorder in La0.5Ca0.5Mn(1 - x)B(x)O3 (B = Fe, Ru, Al and Ga) manganites. AB - We have investigated the influence of B-site doping on the crystal and magnetic structure in La(0.5)Ca(0.5)Mn(1 - x)B(x)O(3) (B = Fe, Ru, Al and Ga) compounds using neutron diffraction, small angle neutron scattering, magnetization and resistivity techniques. The B-site doped samples are isostructural and possess an orthorhombic structure in the Pnma space group at 300 K. A structural transition from orthorhombic to monoclinic is found to precede the magnetic transition to the CE-type antiferromagnetic state in a few of these samples. On doping with Fe, the charge and orbitally ordered CE-type antiferromagnetic state is suppressed, followed by growth of the ferromagnetic insulating phase in 0.02 <= x <= 0.06 compounds. At higher Fe doping in x > 0.06, the ferromagnetic state is also suppressed and no evidence of long range magnetic ordering is observed. In Ru doped samples (0.01 <= x <= 0.05), the ferromagnetic metallic state is favored at T(C)~200 K and T(MI)~125 K and no significant change in T(C) and T(MI) as a function of Ru doping is found. In contrast, with non-magnetic Al substitution for 0.01 <= x <= 0.03, the charge ordered CE-type antiferromagnetic state coexists with the ferromagnetic metallic phase. With further increase in Al doping (0.05 <= x <= 0.07), both CE-type antiferromagnetic and ferromagnetic phases are gradually suppressed. This behavior is accompanied by the evolution of an A-type antiferromagnetic insulating state. Eventually, at higher Al doping (0.10 <= x <= 0.13), this phase is also suppressed and the signature of a spin glass like transition is evident in M(T). Likewise, substitution with Ga is observed to induce similar effects to those described for Al doped samples. The presence of short ranged ferromagnetic ordering has been further explored using small angle neutron scattering measurements in a few of the selected samples. PMID- 21628789 TI - Dynamics in the one-dimensional extended ionic Hubbard model. AB - We investigate single-particle spectral densities and dynamical charge and spin structure factors of the one-dimensional extended ionic Hubbard model in the band insulator regime by using the perturbative continuous unitary transformations method. The one-body staggered potential is considered as the unperturbed part and the hopping term, on-site electron-electron interaction, and the nearest neighbor repulsive interaction are treated as the perturbations. The excitation spectrum of this model was determined in a previous work (Hafez and Jafari 2010 Eur. Phys. J. B 78 323). It was shown that when the intersite interaction is off, there are two antibound state modes and one bound state mode in the singlet channel and two bound state modes in the triplet channel, while for finite values of intersite interaction two bound state modes were found in each channel. Our results for dynamical charge and spin structure factors indicate that only one of two bound/antibound state modes can be probed by electron-energy-loss spectroscopy and inelastic neutron scattering experiments. PMID- 21628792 TI - Tryptamine-induced tryptophanyl-tRNAtrp deficiency in neurodifferentiation and neurodegeneration interplay: progenitor activation with neurite growth terminated in Alzheimer's disease neuronal vesicularization and fragmentation. AB - Tryptophanyl-tRNA synthetase (TrpRS) catalyzes tryptophanyl-tRNAtrp formation. At concentrations exceeding tryptophan, tryptamine inhibits TrpRS. This leads in tryptophanyl-tRNA deficiency and synthesis of aberrant proteins. Tryptamine presents in food and crosses blood-brain barrier. The purpose of this study is to test the hypothesis that tryptamine-induced changes in cell and animal models correlate with Alzheimer's disease (AD) manifestations. Tryptamine prevented growth of human neuroblastoma. Epithelioids recovered growth in tryptamine-free medium, while neuroblasts died. Tryptamine induced epithelioid differentiation forming synaptic vesicles, neuritic contacts, and TrpRS+ axons in stable sublines. A fraction of epithelioids was adhered to satellite cells via trypsin resistant interdigitating junctions. Tryptamine stimulated satellite division and differentiation into neurons, transitional cell variants and neuroblasts able to repopulate. Both tryptamine-inhibited and hypoxia-downregulated TrpRS translocates into cytoplasmic extensions. TrpRS is secreted into extracellular space as a free protein or within vesicles extended from cytoplasm and then pinched-off from plasma membrane of tryptamine-treated cells. Extracellular vesicles fuse in congophilic TrpRS+ plaques in tryptamine-treated culture and AD brain. TrpRS prominent immunoreactivity is associated with plasma and vesicle membranes of satellites and AD brain degenerated neurons. Tryptamine-modified mouse brain expresses amyloid and abnormal filaments in extracellular and neuronal plasma membrane vesicles. Radiolabeled tryptamine, tryptophan and serotonin uptake was 10-fold lower in tryptamine-resistant compared to tryptamine sensitive cells. In both variants, tryptamine uptake exceeded tryptophan uptake within 2-h assuring TrpRS inhibition. Here, tryptophanyl-tRNAtrp deficiency implicates in both neurite growth and termination/collapse. Neurite growth termination prompts TrpRS+ vesicularization. TrpRS+ vesicles contribute in neuronal fragmentation and fibrillar-vesicular congophilic plaques in AD brain. PMID- 21628793 TI - JNK plays a key role in tau hyperphosphorylation in Alzheimer's disease models. AB - Alzheimer's disease (AD) is a major clinical concern, and the search for new molecules to combat disease progression remains important. One of the major hallmarks in AD pathogenesis is the hyperphosphorylation of tau and subsequent formation of neurofibrillary tangles. Several kinases are involved in this process. Amongst them, c-Jun N-terminal kinases (JNKs) are activated in AD brains and are also associated with the development of amyloid plaques. This study was designed to investigate the contribution of JNK in tau hyperphosphorylation and whether it may represent a potential therapeutic target for the fight against AD. The specific inhibition of JNK by the cell permeable peptide D-JNKI-1 led to a reduction of p-tau at S202/T205 and S422, two established target sites of JNK, in rat neuronal cultures and in human fibroblasts cultures. Similarly, D-JNKI-1 reduced p-tau at S202/T205 in an in vivo model of AD (TgCRND8 mice). Our findings support the fundamental role of JNK in the regulation of tau hyperphosphorylation and subsequently in AD pathogenesis. PMID- 21628794 TI - [Joint morphogenesis and development of permanent articular cartilage]. AB - During limb skeletogenesis progenitor mesenchymal cells aggregate at specific times and sites to form continuous precartilaginous condensations. With time the condensations undergo chondrogenesis and give rise to cartilaginous anlagen that exhibit incipient synovial joints at each end. A multitude of factors regulates subdivision into discrete skeletal elements and the formation, organization, morphogenesis and structure of the joints. This review summarizes recent advance of joint morphogenesis and actions of key players of joint and articular cartilage formation. In addition, we would like to discuss possible direction to translate basic research findings towards treatment of joint diseases. PMID- 21628795 TI - [Epidemiology of osteoarthritis in Japan : the ROAD study]. AB - The present study investigated the prevalence of knee osteoarthritis (KOA) and lumbar spondylosis (LS) in the Japanese population, by using a large-scale population-based cohort study named ROAD (Research on Osteoarthritis/osteoporosis Against Disability) . In men and women aged more than 40 years, the prevalence of KOA was 42.6% and 62.4%, respectively, and that of LS was 81.5% and 65.5%, respectively. A total of 25,300,000 people (8,600,000 men and 16,700,000 women) aged more than 40 years could possibly be affected by KOA, and 37,900,000 people (18,900,000 men and 19,000,000 women) by LS. In addition, our study confirmed that factors such as obesity and occupational activities were associated with both KOA and LS. Nutritional factors (vitamin K) were also associated with KOA. The incidence of KOA is significantly related to the increase in the number of metabolic syndrome components such as overweight, hypertension, dyslipidaemia, and impaired glucose tolerance. Finally, we investigated the usefulness of evaluating neuromuscular indices such as hand-grip strength and walking time for predicting the occurrence of a disability. PMID- 21628796 TI - [Recent advance in the genomic study for osteoarthritis]. AB - Osteoarthritis (OA) is a polygenic disease. Genetic factors are involved in its onset and progression. Identification of its genetic factor, i.e., the susceptibility genes for OA is rapidly in progress by using genomic approaches including genome-wide association study. Recent advance in the study of OA susceptibility genes is reviewed. PMID- 21628797 TI - [Analysis of molecular network in chondrocytes by WISH]. AB - One postgenomic strategy used to identify molecular networks functioning in tissue development is microarray analysis of individual cell types or tissues followed by in situ hybridization to identify temporal and spatial gene expression patterns. Compared with microarray analysis, the systematic in situ hybridization database presented here provides more detailed information on the spatial regulation of gene expression and allows identification of discrete clusters of transcribed genes. We created a whole-mount in situ hybridization (WISH) database, containing expression data of transcription factors, cofactors and microRNA expressed in mouse embryos a highly dynamic stage of skeletogenesis. Our approach, WISH provided us new regulators as a critical effector in a myogenic feedback mechanism, tendon development and cartilage homeostasis. PMID- 21628798 TI - [Analysis of chondrocyte differentiation using chondrocyte-specific genetically modified mice]. AB - Knockout mice are used to study physiological function of the gene, while transgenic mice expressing a high level of the transgene are used to examine pharmacological action of the gene. Genes can be deleted not only in chondrocytes, but also at various steps during the process of chondrocyte differentiation in conditional knockout mice. Conditional transgenic mouse system is available in cases in which conventional transgenic mouse lines cannot be established due to early lethality. It is possible that Cre-mediated recombination occurs at loxP sites in osteoblasts of chondrocyte-specific conditional knockout mice, suggesting that bone phenotype should be carefully analyzed and discussed. PMID- 21628799 TI - [Transcriptional regulation in chondrogenesis by Sox9]. AB - To identify a group of transcription factors required for chondrogenesis, several researchers tried to detect a chondrocyte-specific enhancer element of Col2a1 gene. Benoit de Crombrugghe's group finally found out 48bp in the first intron of Col2a1 gene as a chondrocyte-specific enhancer element, and moreover they also concluded that binding of homodimer of Sox9 and homo-or heterodimer of Sox5/Sox6 to this element is indispensable for Col2a1 transcription in chodrocytes. Furthermore, mouse genetic approaches revealed that Sox9, Sox5 and Sox6 are required for chondrogenesis, leading to conclusion that these Sox transcription factors are master regulators in chondrogenesis. Recent studies showed that p300/CBP, Trap230 (med12) , Wwp2, and Med25 are components of transcriptional machinery of Sox9 in chondrogenesis. PMID- 21628800 TI - [Transcriptional regulation of osteoarthritis]. AB - Osteoarthritis (OA) is one of the most common skeletal disorders characterized by cartilage degradation ; however, little was known about the underlying molecular mechanism. Several experimental OA models in mice by producing instability in the knee joints have been developed, and many molecules and signals have been shown to be involved with OA progression through in vivo analyses using mouse OA models. Recently we identified hypoxia-inducible factor-2alpha (HIF2A) as an extensive regulator of the endochondal ossification process. We have shown that HIF2A, which is induced by the NF-kappaB signal, controls OA progression by inducing various target molecules. PMID- 21628801 TI - [Chondrocyte metabolism may be regulated through the interaction with the extracellular matrix]. AB - Articular chondrocytes are the sole type of cell that resides within articular cartilage. The chondrocytes are very sensitive to the matrix around the cells. This is best illustrated by the dramatic change in cell shape and metabolism that occurs during monolayer culture. Although this change, or dedifferentiation, has been known for decades, an underlying mechanism (s) for the change has not yet been determined. Recently, we have reported thatalphavbeta5 integrin is significantly involved in the change through the activation of ERK signaling. This review describes our recent findings about the dedifferentiation of monolayer-cultured chondrocytes. PMID- 21628802 TI - [Novel small compound in combination with cell-sheet technology for articular cartilage regeneration]. AB - Aiming at regeneration of articular cartilage, we have established stable lines of mouse chondrogenic ATDC5 cells expressing green fluorescent protein under the control of type II collagen promoter fused with four repeats of a SOX9 enhancer (COL2A1-GFP) , as a monitoring system for chondrogenic differentiation. A screening of natural and synthetic compound libraries using the system identified some novel compounds. Combined with cell-sheet technology, a novel small compound was applied to the treatment of full-thickness knee cartilage defects in murine and canine models. PMID- 21628803 TI - [Articular cartilage regeneration with synovial mesenchymal stem cells]. AB - Cell transplantation has shown to be a promising strategy to repair cartilage defects. Mesenchymal stem cells derived from synovium have been shown to be a superior cell source for cartilage regeneration to those from other mesenchymal tissues due to their higher rates of colony formation, proliferation potential with autologous serum, and in vitro/vivo chondrogenic potentials. We have found that approximately 60% of synovial mesenchymal stem cells placed on cartilage defects adhered to the defect within 10 min, and the addition of magnesium enhanced this percentage further, which resulted in better cartilage regeneration. Based upon several basic research studies performed in our lab, we have begun the transplantation of synovial stem cells arthroscopically in a clinical study for the treatment of cartilage defects. To date, no adverse events have been reported in the study. Regeneration of cartilage, reduction in defect size and an improvement of symptoms have been obtained in most patients over the last 3 years. PMID- 21628804 TI - [Cell based therapy for articular cartilage injury]. AB - Articular cartilage has the poor capacity for cartilage regeneration, which lead to develop osteoarthritis when cartilage injury occurs. Autologous chondrocyte implantation using three dimensional culture within atelocollagen gel has obtained good clinical results for articular cartilage injury. However, there are two concerns associated with this technique : the number of harvested chondrocyte is limited and arthrotomy is required for implantation of tissue-engineered cartilage. To improve the cell-based articular cartilage treatment, the use of other cell source such as mesenchymal stem cells, and development of less invasive and more efficient cell delivery system should be needed. Translational research will has more and more important implication for clinical application of articular cartilage repair. PMID- 21628805 TI - [Cutting edge on research of cartilage metabolism. Recent progress in bio molecular imaging of articular cartilage]. AB - In the process of cartilage degeneration seen in osteoarthritis, loss of proteoglycan from articular cartilage has been widely accepted as a critical early event, followed by collagen degradation designated as a point of no return. Recent advance in the development of targeted molecular probes and new imaging modalities enabled the detection of qualitative and functional change of articular cartilage. In this paper, we describe the recent progress of bio molecular imaging of articular cartilage including our fluorescent-labeled glycosaminoglycan-binding octaarginine. PMID- 21628806 TI - [MR imaging for the assessment of osteoarthritic knees]. AB - Currently two types of MR imaging are used for the evaluation of osteoarthritic knees. One is to assess the status of knee joint as a whole such as Whole-Organ Magnetic Resonance Imaging Score (WORMS) ; the other is to assess the status of cartilage using specific sequences such as delayed gadolinium enhanced magnetic resonance imaging for cartilage (dGEMRIC) , T2 mapping, and T1 rho. PMID- 21628807 TI - Epidemiology of meningococcal disease in the Panamanian pediatric population, 1998-2008. AB - INTRODUCTION: Worldwide public health authorities report 500,000 cases of invasive meningococcal disease with 50,000 deaths per year and 10-15% of sequelae in people affected. This study describes the epidemiology, microbiology, and clinical presentation of this disease in the Panamanian pediatric population. METHODOLOGY: The discharge of patients with a meningococcal invasive disease diagnosis was reviewed in the statistical database and archives of the Hospital del Nino. RESULTS: A total of 32 discharges with a meningococcal disease diagnosis were reported during the study period (1998-2008). Ninety-one percent (n/N = 29/32) were confirmed as meningitis. The mean age of patients was 4.1 +/- 4.6 years. The incidence in the period of the study was 0.25/100,000. Infants younger than one year old presented the highest incidence rate and number of cases. Four deaths were reported, three of which occurred in the group of 10-14 years and one in the group of 1-4 years. The overall fatality rate was 12.5%. The serogroup of the causative agent, Neisseria meningitidis, was documented in 30 of the 32 cases, with serogroup B the most frequent (66.7%). Ninety-percent (18/20) of serogroup B were isolated in the first five years of study. Serogroup C was identified in 8 of the 12 cases during the period 2004-2008. CONCLUSIONS: The present study showed a change in the epidemiological circulation pattern from serogroup B to serogroup C during the study period. Such epidemiological surveillance data is important in the implementation of preventive measures such as vaccination. PMID- 21628808 TI - Multidrug-resistant typhoid fever: a review. AB - INTRODUCTION: Multidrug-resistant typhoid fever (MDRTF) is defined as typhoid fever caused by Salmonella enterica serovar Typhi strains (S. Typhi), which are resistant to the first-line recommended drugs for treatment such as chloramphenicol, ampicillin and trimethoprim-sulfamethoxazole. Since the mid 1980s, MDRTF has caused outbreaks in several countries in the developing world, resulting in increased morbidity and mortality, especially in affected children below five years of age and those who are malnourished. METHODOLOGY: Two methods were used to gather the information presented in this article. First PubMed was searched for English language references to published relevant articles. Secondly, chapters on typhoid fever in standard textbooks of paediatric infectious diseases and preventive and social medicine were reviewed. RESULTS: Although there are no pathognomonic clinical features of MDRTF at the onset of the illness, high fever ( > 104 degrees F), toxaemia, abdominal distension, abdominal tenderness, hepatomegaly and splenomegaly are often reported. The gold standard for the diagnosis of MDRTF is bacterial isolation of the organism in blood cultures. Ciprofloxacin and ceftriaxone are the drugs most commonly used for treatment of MDRTF and produce good clinical results. CONCLUSION: MDRTF remains a major public health problem, particularly in developing countries. Mass immunization in endemic areas with either the oral live attenuated Typhi 21a or the injectable unconjugated Vi typhoid vaccine, rational use of antibiotics, improvement in public sanitation facilities, availability of clean drinking water, promotion of safe food handling practices and public health education are vital in the prevention of MDRTF. PMID- 21628809 TI - Public knowledge and attitudes towards antibiotic usage: a cross-sectional study among the general public in the state of Penang, Malaysia. AB - INTRODUCTION: Public knowledge and attitudes towards antibiotics play a vital role in the success of the treatment process. This study aimed to assess public knowledge and attitudes toward antibiotic usage which could serve as baseline data for future studies within a government hospital setting in Malaysia. METHODOLOGY: A self-administered cross-sectional survey involving 408 respondents was conducted using a validated questionnaire at an outpatient pharmacy department in Penang Hospital, Malaysia, from February to March 2009. RESULTS: Nearly 55% of the respondents had a moderate level of knowledge. Three quarters of the respondents (76.7%) could correctly identify that antibiotics are indicated for the treatment of bacterial infections. However, 67.2% incorrectly thought that antibiotics are also used to treat viral infections. About 59.1% of the respondents were aware of antibiotic resistance phenomena in relation to overuse of antibiotics. With regard to attitudes, 38% believed that taking antibiotics when having cold symptoms could help them to recover faster, while 47.3% expected antibiotics to be prescribed for common cold symptoms. Age, race and educational level were among the demographic characteristics significantly associated with knowledge and attitudes toward antibiotic use. Poor level of knowledge was found in less than one-third of the respondents whereas more than one-third of the respondents wrongly self-medicate themselves with antibiotics once they have a cold. CONCLUSION: Educational interventions are needed to promote prudent use of antibiotics among the public. PMID- 21628810 TI - Simultaneous detection and subtyping of H274Y-positive influenza A (H1N1) using pyrosequencing. AB - INTRODUCTION: We investigated the frequency of H274Y-positive swine-origin 2009 A (H1N1) influenza virus outbreak in Thailand during May-August 2009. METHODOLOGY: This study sought to find Oseltamivir resistance mutation H274Y by using pyrosequencing. RESULTS: From 8,710 real-time RT-PCR swine-origin 2009 A(H1N1) influenza virus-positive specimens, 100 randomly selected samples identified one such virus with H274Y mutation using pyrosequencing. CONCLUSIONS: The patient probably acquired oseltamivir resistance from natural variation, since he had never received that form of treatment before and recovered from influenza-like symptoms without using anti-influenza drugs. PMID- 21628811 TI - Diagnosis of bacterial vaginosis in cases of abnormal vaginal discharge: comparison of clinical and microbiological criteria. AB - INTRODUCTION: Bacterial vaginosis is a polymicrobial syndrome involving replacement of normal vaginal hydrogen peroxide producing lactobacilli by a variety of mycoplasmas and Gram-negative rods. Bacterial vaginosis has been conventionally diagnosed using Amsel criteria (a clinical method) or Nugent's score (a laboratory method with higher reproducibility). This study was undertaken to compare the diagnostic ability of the Amsel criteria with that of Nugent's score among patients presenting with abnormal vaginal discharge. METHODOLOGY: The study was conducted at the Medical College in Kolkata, India to determine the prevalence of patients with bacterial vaginosis and their demographic profile. Subjects attending the outpatient department presenting with abnormal vaginal discharge were evaluated for the presence of bacterial vaginosis by Amsel criteria and Nugent's score. RESULTS: Prevalence of bacterial vaginosis was 24% by Nugent's score. In comparison, Amsel criteria had sensitivity of 66.67%, specificity of 94.74%, positive predictive value of 80% and negative predictive value of 90%. There was no perfect inter-rater agreement between the Amsel criteria and Nugent's score (Kappa = 0.58). Presence of clue cells correlated best with a positive diagnosis by Nugent's score while the amine test (whiff test) had the lowest correlation. CONCLUSION: Although the Amsel criteria method is a convenient and inexpensive means of diagnosing bacterial vaginosis, it is not always reliable. Alternative reliable and inexpensive diagnostic methods that unify clinical and microbiological parameters, thus increasing sensitivity while retaining specificity, are needed. PMID- 21628812 TI - Salmonella enterica in semi-aquatic turtles in Colombia. AB - INTRODUCTION: Turtles can be hosts of Salmonella enterica serovars which can cause disease both in the animals themselves and in people they come into contact with, especially when the turtles are kept as pets. To investigate the prevalence of Salmonella in turtles in Colombia, we studied animals at a wildlife protection centre. The turtles had either been confiscated or donated to the centre. METHODOLOGY: Detection of Salmonella spp. was conducted in feces samples using bacteriological cultures and polymerase chain reaction to identify genus and serovar. RESULTS: By PCR and culture, 30/110 samples (27%) were positive while by PCR alone eight further samples were positive (total of 38/110 (35%) positive). The most common serovar was S. Enteritidis (26/38 (68%) with only one isolate being S. Typhimurium (3%). Four (11%) samples were positive for both serovars and seven (18%) could only be identified as Salmonella enterica spp. CONCLUSIONS: These results show that turtles in Colombia are commonly infected with Salmonella and are a risk for infection to people who come into contact with them. PMID- 21628813 TI - Comparative study among clinical and commensal isolates of Enterococcus faecalis for presence of esp gene and biofilm production. AB - INTRODUCTION: Because of increasing difficulty in treating enterococcal infections, effort is being devoted to understanding factors that are responsible for causing nosocomial infection, with a focus toward targeting these factors with new therapeutics. Evidence has emerged that the esp gene mediates biofilm formation in vitro, which helps the organism colonize and cause infection. METHODOLOGY: This study was conducted over a four-year period in a tertiary-care hospital. There were 200 clinical pathogenic strains isolated from nosocomial infections and 100 commensals from stool specimens of healthy individuals. The study compared the production of biofilm and detection of the esp gene among clinical and commensal isolates. RESULTS: Among 200 clinical isolates of Enterococcus faecalis 65 (32.5%) isolates were positive for biofilm production and 60 (30%) for the esp gene by PCR. Among 100 commensal isolates, 16 (8%) and 14 (7%) were positive for biofilm formation and the esp gene, respectively. Five clinical and two commensal isolates produced biofilm without any amplification of the esp gene. CONCLUSION: The study shows a significant difference in production of biofilm and presence of the esp gene between clinical and commensal isolates (P < 0.002). Therefore, it can be concluded that biofilm production has an important role in causing nosocomial infection. Although detection of the esp gene correlates with biofilm production, it may not be the only factor determining the formation of biofilm since few isolates produced biofilm without the esp gene. Strains isolated from indwelling medical devices showed high production of biofilm and esp gene. PMID- 21628814 TI - Sequence variation of the HVR1 region of Hepatitis C virus in response to interferon-alpha and ribavirin treatment. AB - INTRODUCTION: Hepatitis C virus (HCV), which is a major cause of liver diseases worldwide, undergoes genetic variation during the course of infection. The aim of this study was to examine sequence variations within the HVR1 region of HCV genotype 4 in infected Saudi patients treated with a combination therapy of interferon-alpha and ribavirin. METHODOLOGY: cDNA of the HVR1 region of HVC-4 from one responder and one non-responder patients was generated, cloned and sequenced. Ten clones were randomly selected and analyzed for changes in nucleotide and amino acid sequences before the start of treatment, and subsequently three and six months after the start of the therapy course. RESULTS: Based on nucleotide and amino acid sequence variations, the HVR1 region is highly sequence variable. In both the responder and the non-responder patients, amino acid sequence variations were observed and a clear distinction between patients was evident. The amino acid changes after the treatment course were different in the responder compared to the non-responder subject. Five amino acids (residues 364 to 367, 381 and 409) were unique in the non-responder patient. CONCLUSION: Considerable amino acid variations were observed in the HVR1 region in both responder and non-responder patients. These findings could have implications for the development of an HCV vaccine as well as treatment protocols for HCV infections. PMID- 21628815 TI - Clinical characteristics and predictors of mortality in hospitalized HIV-infected Nigerians. AB - INTRODUCTION: Human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS) is a chief cause of death in sub-Saharan Africa. In this study, the clinical characteristics and predictors of mortality among hospitalized HIV infected adult Nigerians are reported. METHODOLOGY: The records of 354 patients were reviewed for demographic and clinical characteristics. Predictors of mortality using logistic regression in a retrospective study were also reviewed. RESULTS: A total of 109 (30.8%) males and 245 (69.2%) females participated in the study. The mean age of all participants was 35 +/- 8 years. Median baseline CD4 cell counts and viral load were 91 cells/mm3 and 63,438 copies/ml respectively. There was a total of 123 (34.8%) deaths while 231 (65.2%) patients were discharged home. Tuberculosis (TB) was the most common diagnosis on admission as well as the leading cause of death. Among all subjects, only male gender (adjusted odds ratio [AOR] 4.67, 95% confidence interval [CI]: 2.63-8.29); CD4 cell count <= 200 cells/mm3 (AOR 5.28, 95% CI: 2.99-9.31); length of hospital stay < 3 days (AOR 4.77, 95% CI: 1.35-16.86); and age >= 35 years (AOR 2.43, 95% CI: 1.41-4.19) were predictive of death. CONCLUSION: These findings illustrate the need for early diagnosis of HIV infection, appropriate treatment and prevention of opportunistic infections, and improved access to highly active antiretroviral therapy (HAART). PMID- 21628816 TI - Cyclospora cayetanensis: first report of imported and autochthonous infections in Kuwait. AB - INTRODUCTION: The intestinal coccidian parasites Cryptosporidium spp. and Cyclospora cayetanensis have emerged as significant human pathogens worldwide. The reports of Cyclospora cayetanensis infection in the Middle East are sporadic and no case has been reported from Kuwait. METHODOLOGY: Stool specimens were collected from six individuals presenting with watery diarrhoea of varying degree and severity at the District General hospitals, Kuwait. Four patients were male, two were female and their ages ranged from 5 to 64 years. Three cases were seen among the migrant population from the Indian subcontinent who had recently returned to Kuwait and two cases were seen in the local population with no history of travel abroad. The stool smears were stained with modified acid-fast stain and examined under ultraviolet (UV) fluorescence illumination. RESULTS: Cyclospora cayetanensis oocysts with variable staining characteristics were detected by microscopy. Two of the cases were also associated with other enteric pathogens. Clinical suspicion of Cyclospora infection was not recorded for any of the cases. All patients showed remarkable symptomatic and parasitologic improvements upon treatment with trimethoprim/sulfamethoxazole. CONCLUSION: Cyclospora spp. are important aetiological agents of diarrhoea in the Middle East. An awareness of the parasitic infection and use of appropriate diagnostic modalities are essential to elucidate the clinical and epidemiological significance of the parasitosis in this geographic area. PMID- 21628817 TI - In vitro activity of azithromycin in Salmonella isolates from Pakistan. AB - INTRODUCTION: Enteric fever is caused by Salmonella enterica serovars Typhi and Paratyphi A, B and C. It is a significant public health issue in Pakistan, which is exacerbated by a high level of resistance some isolates display to drugs routinely used in treatment. Azithromycin may be a treatment option for such isolates. METHODOLOGY: We determined the minimum inhibitory concentrations (MICs) of Salmonella Typhi and Paratyphi isolates against azithromycin in an attempt to gauge its feasibility as a therapeutic option. The MICs were also compared with corresponding disc diffusion zone sizes to see if there was consistency between the two tests. We tested 45 Salmonella enterica isolates using E-tests for MIC detection and azithromycin discs with a concentration of 15ug/ml for disc diffusion testing. RESULTS: Salmonella Typhi, Salmonella Paratyphi A, and Salmonella Paratyphi C isolates demonstrated MICs of 2-12mg/L against azithromycin, suggesting that the antibiotic could be used for therapeutic purposes. For Salmonella Paratyphi B, the MICs were 2-48 mg/L. The higher MIC indicates a need for caution when considering use of azithromycin for Salmonella Paratyphi B infections without first testing for the MIC. There was a close correlation between MICs and zone sizes which was statistically significant. CONCLUSIONS: Our results indicate azithromycin is a potential therapeutic option for enteric fever. Standardized laboratory testing methods and interpretation for azithromycin against Salmonella enterica would allow laboratories to report upon this antibiotic with confidence. PMID- 21628818 TI - The sale of antibiotics without prescription in pharmacies in Damascus, Syria. AB - INTRODUCTION: Overuse of antibiotics has contributed to the development of organism resistance. The acquisition of antibiotics without prescription by the general population seems to be common practice in pharmacies of Damascus, Syria. This study aimed to determine the proportion of pharmacies dispensing antibiotics without medical prescription and without seeing the patient. METHODOLOGY: A cross sectional study involving a sample of 224 pharmacies was conducted in Damascus. To obtain antibiotics without medical prescription, the investigators posed as individuals who had a sister with symptoms of sinusitis. RESULTS: From 200 pharmacies visited, 87% agreed without insistence from the investigator to sell antibiotics without prescription. This figure increased to 97% when the investigators who were at first denied antibiotics insisted on having the antibiotics. CONCLUSION: Dispensing of antibiotics is high in Damascus despite federal regulations. Health education programs should be directed to pharmacies and also to the population. PMID- 21628819 TI - Fatal case of Clostridium perfringens enteritis and bacteraemia in South Africa. AB - Clostridium perfringens is an important anaerobic pathogen causing foodborne and non-foodborne gastrointestinal diseases in humans and animals. This pathogen is also the more common Clostridium species associated with bacteraemia. We report on a fatal case of C. perfringens infection in an adult with type 2 diabetes. PMID- 21628820 TI - Case report on the development of a brucellosis-related epidural abscess. AB - Brucellosis is an endemic disease in developing countries. The most commonly observed complications include bone-joint involvement, particularly sacroiliitis and spondylitis. Epidural abscesses caused by brucellosis are a rare complication. We describe the case of a 33-year-old man presenting with high fever, back pain, and weakness. At physical examination, the patient was found to be paraparetic. At thoracic MRI, spondylodiscitis and epidural abscess with significant cord compression were observed. In laboratory examinations, Rose Bengal and tube agglutination tests were positive in patient's serum for brucellosis, and specific antibiotherapy was initiated. Total laminectomy was performed and the abscess was aspirated. The biopsy sample was consistent with chronic non-specific inflammation in acute abscess. No growth was detected in the abscess or blood cultures. Following surgery, medical treatment was initiated and, at six weeks' follow-up, clinical and MRI findings indicated that he had recovered. The diagnosis of spinal epidural abscess due to brucellosis should be considered among differential diagnoses in endemic regions. Early diagnosis and specific treatment are important to prevent later complications. PMID- 21628821 TI - Isolated cutaneous aspergillosis in an acute lymphoblastic leukemia patient after allogeneic stem cell transplantation. AB - Cutaneous aspergillosis is very rare and occurs predominantly in immunocompromised patients including transplant recipients. We report a 26-year old male with acute lymphoblastic leukemia who developed cutaneous aspergillosis after undergoing combined immunosuppressive treatment including corticosteroid, cyclosporine A, mychophenolate mofetil and mesenchymal stem cells for steroid refractory skin acute graft versus host disease after myeloablative haematopoietic stem cell transplantation. The patient was treated with oral voriconazole therapy and recovered partially. PMID- 21628822 TI - Low prevalence of transfusion-transmissible infections among voluntary blood donors in South India. PMID- 21628823 TI - Asymmetrical lower extremity loading after ACL reconstruction: more than meets the eye. AB - Sports fans know that movement patterns are important for athletic performance. Similarly, clinicians know that addressing abnormal movement patterns after an anterior cruciate ligament (ACL) reconstruction is important for a successful return to sport. The kinematic (motion) component of movement patterns is more easily observed than the kinetic (forces) component, thus more commonly addressed in ACL reconstruction rehabilitation. Ignoring the kinetic component, though, could impede a successful return to sport. Asymmetrical lower extremity loading has been reported in a variety of activities following ACL reconstruction, and may contribute to both short- and long-term consequences. It is important that clinicians become aware of the potential for asymmetrical lower extremity loading to affect patient outcomes and for researchers to enlarge the body of knowledge. PMID- 21628824 TI - Return to sport: when should an athlete return to sport after an ACL surgery? AB - A torn anterior cruciate ligament (ACL), followed by reconstruction surgery, can be devastating for an athlete. Advances in physical therapy now help athletes improve rapidly during the early period after surgery. However, guidelines on how to determine if it is safe to return to sport are more general and vary widely. Athletes and the people who care about and for them need reliable and valid methods to determine when they are ready to return to sport. A study published in the June 2011 issue of JOSPT provides new insight and evidence-based tools to help answer this question. PMID- 21628825 TI - Computer simulation of pectoralis major muscle strain to guide exercise protocols for patients after breast cancer surgery. AB - STUDY DESIGN: Descriptive study. OBJECTIVES: To quantify and rank the order of strain (length change in proportion to the resting length) of 3 portions of the pectoralis major (PM) muscle during various exercises. BACKGROUND: A biomechanical foundation on which to base exercise prescriptions for patients after breast cancer surgery is lacking. METHODS: An interactive, 3-D, computer graphic simulation system, developed to study biomechanical properties of the musculoskeletal system, was used to simulate movements of the glenohumeral, scapulothoracic, and scapuloclavicular joints of the shoulder, and to estimate strain in 3 portions of the pectoralis major (PM) muscle throughout the motions. The computed tomography scans of 2 male cadavers and literature review formed the basis for the estimations used in the model. Strains in the clavicular, midsternum, and abdominal regions of the PM were expressed as percent strain: [(change in muscle length/resting length) * 100]. Exercise motions were based on PM muscle anatomy and published breast cancer rehabilitation protocols. RESULTS: Strains of the PM regions ranged from -21% shortening of the clavicular region during flexion to 55% lengthening of the abdominal region during the overhead stretch. Strain between adjacent regions was most uniform for the movement of abduction with external rotation, and least uniform with flexion. CONCLUSION: PM muscle lengthening estimates were not linearly proportioned to shoulder joint motions, and varied for 3 portions of the PM. This information may help clinicians and researchers to estimate lengthening of PM portions throughout measurable shoulder motions. PMID- 21628826 TI - Conflicting dermatome maps: educational and clinical implications. AB - SYNOPSIS: Sensory testing is a common noninvasive method of evaluating nerve function that relies on the knowledge of skin dermatomes and sensory fields of cutaneous nerves. Research to determine the extent of the dermatomes was conducted in Europe during the late nineteenth and early twentieth centuries. Experiments performed on cadavers, monkeys, and human patients prior to 1948 resulted in the creation of similar but somewhat different dermatome maps. A radically different map with long, swirling dermatomes was produced by Keegan and Garrett in 1948. This map was derived largely by examining compression of dorsal nerve roots by vertebral disc herniation. The maps appearing in textbooks are inconsistent. Some books show a version of the early maps, some show the Keegan and Garrett map, and others show maps that are not consistent with either. The purpose of this paper is to discuss the history of dermatome maps, including the experimental procedures by which each was obtained, and to relate the early maps to those found in textbooks commonly used in healthcare education programs. The paper discusses the significance of these maps as used for clinical diagnosis and the need for further research. PMID- 21628827 TI - Occult hypermobility of the craniocervical junction: a case report and review. AB - STUDY DESIGN: Resident's case problem. BACKGROUND: Patients often present to physical therapists with chief complaints of neck pain, occipital headache, and dizziness associated with a past history of cervical spine injury. These symptoms may be associated with various cervical spine conditions, including craniocervical junction (CCJ) hypermobility. DIAGNOSIS: This report reviews the history, physical exam, and diagnostic imaging findings of a patient with the above symptoms. This patient, who had a history of multiple cervical spine injuries, was examined with 2 manual therapy provocative tests: the Sharp-Purser test, which is intended to stress the transverse ligament and odontoid, and the modified lateral shear test, which is intended to stress the alar ligaments. The lateral shear test was perceived as demonstrating excessive mobility and a soft end feel, with a "shift" of C1 on C2. Stress cervical radiographs, obtained using open-mouth projections in neutral, left, and right cervical lateral flexion, revealed a 3-mm lateral offset of the right lateral mass of C1 on C2. MRI evaluation of the lower cervical spine did not reveal any significant disc derangement; however, images of the soft tissues of the craniocervical junction were not obtained. Based on the examination and imaging studies, the patient was determined to have a previously undiagnosed hypermobility of the atlantoaxial joint. DISCUSSION: The patient was advised to avoid rotational manipulation and end range lateral flexion stretching exercises. Axial traction manipulation techniques, midrange stabilization exercises, and postural advice appeared to provide good relief of symptoms. Physical therapists should consider the possibility of CCJ hypermobility in the frontal plane when examining the cervical spine in patients with chronic neck pain, headache, and a past history of trauma. The lateral shear test and stress radiography may provide simple screening tests for occult CCJ hypermobility; however, the reliability and validity of these tests is lacking. Further research on diagnosis and management of CCJ hypermobility is warranted. LEVEL OF EVIDENCE: Differential diagnosis, level 4. PMID- 21628828 TI - Posttransplant distal limb syndrome. AB - The patient was a 45-year-old woman who was referred to a physical therapist and a physiatrist in a rehabilitation medicine department for evaluation and treatment of severe bilateral lower leg, ankle, and foot pain. The patient's past medical history was significant for sickle cell disease and she had undergone an allogeneic stem cell transplant 4 months prior with Sirolimus prescribed to prevent rejection. Magnetic resonance imaging of both lower legs revealed extensive bone marrow edema, as well as soft tissue swelling about the lower legs and ankles. These findings, along with the patient's presentation (constant bilateral pain and erythema of the lower legs within 6 months of transplantation) were found to be consistent with an atypical condition called posttransplant distal limb syndrome. PMID- 21628829 TI - Cystic fibrous dysplasia of the humerus. AB - The patient was a 39-year-old woman with a 3-month history of worsening right distal humerus pain that was insidious in onset. At the time of the initial physical therapy visit, the patient complained of weakness and numbness throughout the entire right upper extremity, with an inability to perform daily activities. Magnetic resonance imaging revealed extensive bone marrow signal abnormalities extending from the proximal humeral epiphysis to the distal humeral diaphysis. Further evaluation with radiographs and computed tomography revealed multiple lytic lesions through the humerus. Tissue biopsy confirmed a diagnosis of cystic fibrous dysplasia. PMID- 21628831 TI - Long-term treatment with eicosapentaenoic acid ameliorates myocardial ischemia reperfusion injury in pigs in vivo. -Involvement of Rho-kinase pathway inhibition . AB - BACKGROUND: Eicosapentaenoic acid (EPA), the major n-3 fatty acid in fish oil, exerts cardioprotective effects against ischemic heart disease; however, the detailed mechanisms remain to be elucidated. Rho-kinase plays an important role in the pathogenesis of cardiovascular diseases including ischemia-reperfusion (I/R) injury. Thus, the hypothesis that long-term EPA treatment ameliorates myocardial I/R injury through Rho-kinase pathway inhibition in pigs in vivo was investigated. METHODS AND RESULTS: Male pigs were treated with either a control chow or EPA (600.mg.kg-1.day-1) for 3 weeks (n=8 each) and were subjected to myocardial ischemia by 90-min occlusion of the left circumflex coronary artery and subsequent 60-min reperfusion. The EPA group had an increased EPA level in red blood cells (4.4 +/- 0.3mol%). The EPA treatment significantly ameliorated myocardial I/R injury, including regional wall motion abnormality (EPA 5.3 +/- 3.6 vs. control 35.1 +/- 3.8 unit, P<0.0001), left ventricular ejection fraction (EPA 43 +/- 9% vs. control 32 +/- 7%, P<0.05), occurrence of ventricular arrhythmias (EPA 181 +/- 73 vs. control 389 +/- 51 events, P<0.0001) and histological accumulation of inflammatory cells (P<0.01). Importantly, the EPA treatment significantly inhibited myocardial Rho-kinase activity (assessed by the extent of the myosin-binding subunit phosphorylation) (EPA 0.47 +/- 0.11 vs. control 0.77 +/- 0.14, P<0.05) and preserved myocardial eNOS activity (EPA 0.56 +/- 0.13 vs. control 0.23 +/- 0.07, P<0.01) with a significant correlation noted between them. CONCLUSIONS: Long-term treatment with EPA ameliorates I/R injury partly through Rho-kinase pathway inhibition in vivo. PMID- 21628832 TI - Impact of hinge motion on in-stent restenosis after sirolimus-eluting stent implantation. AB - BACKGROUND: A tortuous lesion with hinge motion is reportedly a risk factor for in-stent restenosis (ISR) after bare metal stent implantation. Sirolimus-eluting stents (SESs) implantation has dramatically reduced ISR. However, SES is a closed cell design stent, which has low conformability and flexibility. Several studies have reported a relationship between tortuous lesions and stent fracture, which is one of the causes of ISR. The efficacy of SES in a tortuous lesion with hinge motion has not been fully evaluated. The aim of this study was to investigate the relationship between tortuous lesions and ISR after SES implantation. METHODS AND RESULTS: Three year clinical follow-up data after SES implantation, including 6-9 month scheduled follow-up coronary angiography in 399 consecutive patients with 537 lesions, were obtained. Deltaangle was defined as the difference in the angle of the target lesion between the diastole and systole before the procedure. The incidence of ISR was 8.2%. The mean maximal angle and Deltaangle were larger in the ISR group (47 +/- 22 degrees vs. 37 +/- 21 degrees , P=0.004 and 20 +/- 13 degrees vs. 13 +/- 10 degrees , P < 0.0001, respectively). Independent predictors of ISR were Deltaangle, hemodialysis, aortic ostium stenting, and diabetes mellitus. Hinge motion-associated ISR (Deltaangle >= 16 degrees ) occurred in 28 lesions: stent fracture in 9, stent recoil in 6, edge injury in 10, and others in 3. CONCLUSIONS: The amount of hinge motion in tortuous lesions should be considered in the selection of drug eluting stent types. PMID- 21628834 TI - Less invasive approach to the assessment of coronary artery disease. Morphological vs. physiological paradigm-. PMID- 21628833 TI - Impaired subendocardial wall thickening and post-systolic shortening are signs of critical myocardial ischemia in patients with flow-limiting coronary stenosis. AB - BACKGROUND: The early diagnosis of myocardial ischemia is still challenging. The aim of the present study was to determine whether subendocardial hypokinesis and post-systolic contraction could be early markers of myocardial ischemia. METHODS AND RESULTS: Thirty-one consecutive patients with flow-limiting severe coronary stenosis but without visually abnormal left ventricular wall motion underwent quantitative echocardiography. Myocardial strain was measured using layer-by layer analysis in severely hypoperfused segments. Radial strain (RS) was measured in the subendocardial, subepicardial, and total wall (innerRS, outerRS, and totalRS, respectively). Circumferential strain (CS) was also measured as 3 separate layers: subendocardial, mid-layer, and subepicardial layers (innerCS, midCS, and outerCS, respectively). Post-systolic shortening (PSS) was defined as the peak strain after end systole, and post-systolic strain index (PSI) was calculated as PSS divided by end-systolic strain. TotalRS was similar between ischemic and normally perfused segments, but innerRS and inner/outer RS ratio were significantly smaller in the ischemic segments than in corresponding segments in healthy subjects. Receiver operating characteristic analysis identified an optimum cut-off for PSI of 0.6. The combined criteria of inner/outer RS ratio <1.0 and PSI >0.6 achieved 95% specificity for the presence of flow-limiting stenosis. CONCLUSIONS: Combined assessment of both subendocardial contractile impairment and PSS is very useful in identifying a severely hypoperfused left ventricular wall even without visual wall motion abnormality. PMID- 21628835 TI - Dysfunctional high-density lipoprotein and the potential of apolipoprotein A-1 mimetic peptides to normalize the composition and function of lipoproteins. AB - Although high-density lipoprotein-cholesterol (HDL-C) levels in large epidemiological studies are inversely related to the risk of coronary heart disease (CHD), increasing the level of circulating HDL-C does not necessarily decrease the risk of CHD events, CHD deaths, or mortality. HDL can act as an anti or a pro-inflammatory molecule, depending on the context and environment. Based on a number of recent studies, it appears that the anti- or pro-inflammatory nature of HDL may be a more sensitive indicator of the presence or absence of atherosclerosis than HDL-C levels. The HDL proteome has been suggested to be a marker, and perhaps a mediator, of CHD. Apolipoprotein A-1 (apoA-I), the major protein in HDL is a selective target for oxidation by myeloperoxidase, which results in impaired HDL function. Improving HDL function through modification of its lipid and/or protein content maybe a therapeutic target for the treatment of CHD and many inflammatory disorders. HDL/apoA-I mimetic peptides may have the ability to modify the lipid and protein content of HDL and convert dysfunctional HDL to functional HDL. This review focuses on recent studies of dysfunctional HDL in animal models and human disease, and the potential of apoA-I mimetic peptides to normalize the composition and function of lipoproteins. PMID- 21628836 TI - Morphological and quantitative analysis of vascular wall and neointimal hyperplasia after coronary stenting: comparison of bare-metal and sirolimus eluting stents using optical coherence tomography. AB - BACKGROUND: It has been suggested that sirolimus-eluting stents (SES) provoke a more sustained inflammatory response (IR) in neointimal hyperplasia (NIH). The purpose of this study was to compare morphological vessel characteristics, including post-stent IR in NIH, between patients with SES and bare metal stents (BMS) using optical coherence tomography (OCT). METHODS AND RESULTS: Thirty-seven patients underwent OCT at their post-stent follow-up. OCT signal-intensity deviation (normalized standard-deviation; OCT-NSD) values in NIH were compared between the 2 groups. In addition, the serum concentration of high-sensitivity C reactive protein (hs-CRP) was measured. Stent-malapposition rate (1.78% vs. 0.7%; P = 0.016), uncovered stent-strut rate (16% vs. 3.7%; P = 0.0002), peri-stent ulcer like appearance (PSUA; 50% vs. 0%; P = 0.006) were all significantly higher in the SES group than in the BMS group, respectively. The OCT-NSD value was also significantly higher in the SES group than in the BMS group (0.213 +/- 0.005 vs. 0.198 +/- 0.005; P < 0.001), as was the hs-CRP level (2.54 +/- 1.89 vs. 0.64 +/- 0.3 mg/L; P = 0.0006). In addition, a significant positive correlation was found between hs-CRP and OCT-NSD (r = 0.471; P = 0.0025). CONCLUSIONS: PSUA-morphology was specific in the SES group, and higher levels of OCT-NSD and hs-CRP after SES implantation suggest sustained IR in NIH compared with following BMS implantation. These different characteristics may be some of the background that promotes thrombus formation as a late-stage post-stent complication of SES. PMID- 21628837 TI - Characteristics and predictors of drug-eluting stent thrombosis: results from the multicenter 'Korea Stent Thrombosis (KoST)' registry. AB - BACKGROUND: Previous studies have reported possible predictors of drug-eluting stent thrombosis (ST), but data for Asians are relatively limited. This study was performed to elucidate clinical predictors of ST in Koreans. METHODS AND RESULTS: From May 2003 to May 2007, consecutive patients presenting with ST were enrolled from 10 cardiovascular centers in Korea. They were compared with 2,192 controls (3,223 lesions) who had received percutaneous coronary intervention with at least 6 months of follow-up without ST. On multivariate analysis, acute myocardial infarction (AMI) as initial diagnosis, drug-eluting stents (DES) in-stent restenosis (ISR), low ejection fraction (EF), small stent diameter, left anterior descending artery intervention, and young age were independent predictors of total ST. When divided into early (ST within 30 days of index procedure) and delayed ST (ST after 30 days of index procedure), low EF, small stent diameter, DES ISR and AMI as initial diagnosis were universal risks for both early and delayed ST. The time from antiplatelet agent discontinuation to ST occurrence was significantly shorter in late compared with very late ST. CONCLUSIONS: Predictors of ST may be slightly different for early vs. delayed ST. However, low EF, small stent diameter, DES ISR lesion, and AMI as initial diagnosis were universal risk factors for both early and delayed ST cases. The relationship between antiplatelet agent discontinuation and ST occurrence seems stronger in late compared with very late ST. PMID- 21628838 TI - Diet-induced lipid accumulation in liver enhances ATP-binding cassette transporter g5/g8 expression in bile canaliculi. AB - The ATP-binding cassette half-transporters Abcg5 and Abcg8 promote the secretion of neutral sterols into bile. Studies have demonstrated the diet-induced expression of these transporters in liver, but precisely where this occurs remains to be elucidated. This study investigated the changes in the expression of these transporters in bile canaliculi in cholesterol-loaded livers. Mice were fed either a standard (SD) diet or a high-fat and high-sucrose (HF/HS) diet for 3 weeks. Bile canaliculi proteins and cryosections were prepared from the liver, and the protein levels and distribution of Abcg5/Abcg8 were determined. The high calorie diet induced a marked accumulation of lipids in mouse liver. Protein levels of Abcg5 and Abcg8 in bile canaliculi were significantly increased by the HF/HS diet compared to the SD diet. No significant differences in Abca1, Abcb4 (Mdr2), Abcb11 (Bsep), or Abcc2 (Mrp2) levels were observed. Immunohistochemical analyses confirmed that these increases occurred in bile canaliculi. These results suggest that diet-induced lipid loading of the liver causes a significant increase in the expression of Abcg5 and Abcg8 in bile canaliculi. PMID- 21628839 TI - Cardio-ankle vascular index (CAVI) as a novel indicator of arterial stiffness: theory, evidence and perspectives. AB - The cardio-ankle vascular index (CAVI) is a new index of the overall stiffness of the artery from the origin of the aorta to the ankle. The most conspicuous feature of CAVI is its independence of blood pressure at the time of measurement.CAVI increases with age and in many arteriosclerotic diseases, such as coronary artery disease, carotid arteriosclerosis, chronic kidney disease and cerebrovascular disease, and is related to many coronary risk factors, such as hypertension, diabetes mellitus, dyslipidemia and smoking. Furthermore, CAVI decreases by controlling diabetes mellitus and hypertension, and also by abstaining from smoking. This suggests that CAVI is a physiological surrogate marker of athero- or arteriosclerosis, and also might be an indicator of lifestyle modification.Recently, it has been reported that CAVI and several left ventricular functions are co-related, suggesting a connection between the heart muscle and vascular function.This review covers the principles of CAVI and our current knowledge about CAVI, focusing on its roles and future outlook. PMID- 21628841 TI - Disability-adjusted life years (DALY) for cancer in Japan in 2000. AB - BACKGROUND: We used disability-adjusted life years (DALY) to estimate the cancer burden in Japan for the year 2000. METHODS: We estimated years of life lost (YLL) by using mortality data and years lived with disability (YLD) by using incidence data. The DALY for cancer was calculated as the sum of YLL and YLD. RESULTS: For all cancers combined, 2 733 884 years of DALY were estimated in men and 2 091 874 years were estimated in women. Among men, stomach and lung cancers accounted for the largest proportions of DALY, followed by liver cancer and colorectal cancer. Among women, the greatest contributors to DALY were stomach, colorectal, breast, and lung cancers. CONCLUSIONS: The national cancer burden in Japan was expressed in terms of DALY, which might be useful in assessing future changes with respect to mortality and morbidity in Japan. PMID- 21628842 TI - Maternal diet and selenium concentration in human milk from an Italian population. AB - BACKGROUND: Low selenium (Se) status is associated with several diseases. International organizations have proposed intakes of Se for general populations, including infants. Studies of the association of Se concentration in breast milk and maternal diet have yielded inconsistent results. We evaluated the relation between the intake of food items during pregnancy and Se concentration in human milk after delivery and compared infant intake of Se from breast milk with the recommended intakes. METHODS: This cross-sectional study was part of the baseline assessment of a prospective cohort of Italian mother-child pairs enrolled in 1999 2001. Se concentration was measured in the milk of 100 women included in the cohort and correlated with the intake of food items during pregnancy and lactation as reported in a food frequency questionnaire. RESULTS: Among foods consumed in pregnancy, only eggs had a positive, but weak, correlation with Se concentration in milk (r = 0.20, P = 0.04). Fish intake during lactation was also weakly correlated with Se in milk (r = 0.21, P = 0.04). Se content of breast milk in our population was lower than that noted in other international studies; however, very few children who were exclusively breastfed were estimated not to have met the recommended Se intake. CONCLUSIONS: Future research should aim to verify whether infants in this part of Italy still meet the recommended nutrient intake of Se and to assess the influence of the concurrent diet of lactating mothers on the Se content of their milk. PMID- 21628843 TI - Duration of prion disease is longer in Japan than in other countries. AB - BACKGROUND: Prion diseases are untreatable, progressive, and fatal brain disorders that occur worldwide, and the annual incidence rate is approximately 1 case per 1 million people. The duration of these diseases in Japan is unclear. METHODS: Based on data from 1 April 1999 through 4 September 2008 provided by the Japanese Creutzfeldt-Jakob disease (CJD) surveillance program, we analyzed disease duration and its relationship with clinical features. Duration was assumed to be the time from disease onset to death. RESULTS: Evaluation by the surveillance committee indicated that during the observed period 1128 individuals received a diagnosis of prion disease and were registered in the surveillance program. Mean disease duration in the 855 patients who died was 17.4 months. Overall, 46.0% of patients died within 1 year and 77.2% died in less than 2 years. Among those with sporadic Creutzfeldt-Jakob disease, which represented 77.0% of cases, mean disease duration was 15.7 months, while that of patients surveyed by the European Creutzfeldt Jakob Disease Surveillance Network (EUROCJD) was only 5 months. CONCLUSIONS: Disease duration among Japanese with prion diseases was much longer than that of patients in Western countries conducting surveillance of prion diseases. This finding suggests that the characteristics of the system for providing life-sustaining treatment for patients with fatal, progressive diseases in Japan are related to the longer duration of these illnesses. PMID- 21628844 TI - Attenuated lung fibrosis in interleukin 6 knock-out mice after C-ion irradiation to lung. AB - There is a great deal of evidence that a cyclic cascade of inflammatory cytokines, together with the activation of macrophages, is initiated very early after irradiation to develop lung fibrosis in a late phase. To understand the persistent effects of cytokines, the cytokine gene of knock out or transgenic mouse is one of the useful tools. In this study, we evaluated a role of a key molecule, interleukin-6 (IL-6), in the late-phase inflammatory response and subsequent fibrotic changes after irradiation using wild-type (WT) and IL-6 knock out (IL-6 KO) mice. The mice underwent thoracic irradiation with 10 Gy of C-ion beam or sham-irradiation and were examined by histology. Immunoreactivity for IL 6 was induced at the site of bronchiolar epithelium, in pneumocytes and in monocytes by C-ion irradiation. At 24 weeks after irradiation, the infiltration of macrophages, detected by positive immunohistological staining with Mac3 antibody, was observed in alveolar spaces both in WT and IL-6 KO mice. The thickening of bronchiolar and alveolar walls exhibited in WT mice, but not KO mice, and fibrotic changes detected by Masson-Trichrome staining, were observed only in the lungs of WT mice, while it was attenuated in IL-6 KO mice. These results indicated that IL-6 might not be essential for activating macrophages in the late phase, but plays an important role for fibrotic changes of the alveolar wall after irradiation. PMID- 21628847 TI - [Basophils in skin inflammation]. AB - Basophils represent less than 1% of peripheral blood leukocytes. Under physiological conditions, basophils principally circulate in peripheral blood, while mast cells reside in peripheral tissues. Like mast cells, they express the high-affinity IgE receptor on their cell surface and release chemical mediators. Because of morphological and functional similarities, basophils have long been considered to be redundant "circulating mast cells" and minor (probably negligible) players in inflammation. Mouse and human basophils cannot be stained in routinely processed histological specimens, and thus, our understanding of tissue basophils in allergic inflammation had been limited. However, recent studies in mice have revealed that basopihls play non-redundant roles from mast cells. Basophils function as a source of IL-4, IL-13, and CCL22, thereby contributing Th2 immunity. They are also capable of presenting antigens. Basophils are essential for the development of IgE-mediated chronic allergic skin inflammation in mice. Recent immunohistochemical studies with an basophil specific antibody revealed that, in humans, varying numbers of basophils infiltrate skin lesions of inflammatory diseases, such as atopic dermatitis, urticaria, prurigo, and eosinophilic pustular folliculitis. Basophils may play important roles in a variety of inflammatory skin diseases than previously thought. PMID- 21628845 TI - Radiation-generated short DNA fragments may perturb non-homologous end-joining and induce genomic instability. AB - Cells exposed to densely ionizing radiation (high-LET) experience more severe biological damage than do cells exposed to sparsely ionizing radiation (low-LET). The prevailing hypothesis is that high-LET radiations induce DNA double strand breaks (DSB) that are more complex and clustered, and are thereby more challenging to repair. Here, we present experimental data obtained by atomic force microscopy imaging, DNA-dependent protein kinase (DNA-PK) activity determination, DNA ligation assays, and genomic studies to suggest that short DNA fragments are important products of radiation-induced DNA lesions, and that the lengths of DNA fragments may be significant in the cellular responses to ionizing radiation. We propose the presence of a subset of short DNA fragments that may affect cell survival and genetic stability following exposure to ionizing radiation, and that the enhanced biological effects of high-LET radiation may be explained, in part, by the production of increased quantities of short DNA fragments. PMID- 21628848 TI - [Langerhans cell and HIV]. AB - Heterosexual transmission of HIV is the most common mode of infection in the global HIV epidemic. In the absence of an effective vaccine, there is an urgent need for additional strategies to prevent new HIV infections. Evidence from a variety of investigations, including epidemiologic studies on sexual transmission, in vivo studies in rhesus monkey, and ex vivo studies using human explant models, indicate that CD4/CCR5-mediated de novo infection of Langerhans cells (LCs) is a major pathway involved in sexual transmission of HIV (LCs primary gate keeper model). However, it has been recently revealed that Langerin (a C-type lectin receptor) expressed on LC inactivate HIV. Thus, there may be multiple ways by which HIV interacts with LCs in the genital mucosa. In light of the current HIV infection rates in heterosexuals and the absence of a prophylactic vaccine, prevention strategies, such as topical microbicides that block sexual transmission of HIV, are urgently needed. This review focuses on the recent advances regarding the role of LCs in heterosexual transmission of HIV, and the relationship between the LCs primary gate keeper model and current prevention strategies worldwide. PMID- 21628849 TI - [Dynamic cooperation between epidermal barriers and Langerhans cells]. AB - Skin is the structure that covers our body and protects it from not only the entry of pathogens or allergens but also from the leakage of water, solutes or nutrients. These outside-in and inside-out skin barrier functions are dependent on the epidermis, a stratified epithelial cellular sheet. While mucus covers the epidermis in fish and amphibian tadpoles, terminally differentiated cornified cellular sheets called stratum corneum (SC) constitute the outermost epidermal barrier in amphibian adults, reptiles, birds and mammals. Beneath the mucus or SC, apical paracellular spaces of epidermal cells are sealed with tight junctions (TJs) that might limit paracellular leakage of water and electrolytes to maintain fluid homeostasis. We have recently reported in mice that Langerhans cells (LCs) elongate their dendrites to penetrate through epidermal TJs upon activation and uptake antigens from extra-TJ environment. During antigen uptake, new TJs are formed between keratinocytes and LC dendrites to maintain the integrity of epidermal TJ barriers. To understand the epidermal barrier system and its deficiency observed in human skin diseases, we need to re-evaluate human epidermal barrier as a composite barrier consisting of SC and TJs and to investigate the molecular mechanism and immunological consequences of the extra TJ antigen uptake activity of LCs. PMID- 21628850 TI - [Therapy for skin disease using bone marrow cells]. AB - Attempts to treat congenital protein deficiencies using bone marrow-derived cells have been reported. These efforts have been based on the concepts of stem cell plasticity. We aimed to clarify whether bone marrow transplantation (BMT) treatment can rescue epidermolysis bullosa (EB) caused by defects in keratinocyte structural proteins. BMT treatment of adult collagen XVII (Col17) knockout mice induced donor-derived keratinocytes and Col17 expression associated with the recovery of hemidesmosomal structure and better skin manifestations, as well improving the survival rate. Furthermore, human cord blood CD34+ cells also differentiated into keratinocytes and expressed human skin component proteins in transplanted immunocompromised mice. The current conventional BMT techniques have significant potential as a systemic therapeutic approach for the treatment of human EB. PMID- 21628851 TI - [Cutaneous lymphoma and chemokine]. AB - The expression pattern of chemokines and chemokine receptors is specific to certain organs and cells. Therefore, chemokines are important to elucidate the mechanism of organ-specific human diseases such as cutaneous lymphoma, characterized by proliferation of clonally expanded lymphocytes in skin without detectable systemic involvement. The most popular type of cutaneous lymphoma is T cell lymphoma, including mycosis fungoides and Sezary syndrome. We have reported that CCL17, CCL27, CCL11, and CCL26 are involved in progression of these diseases. The above chemokines are highly expressed in the lesional skin and serum levels of the chemokines are elevated as the disease progressed. Moreover, CXCL9 and CXCL10 are associated with epidermotropism of tumor cells, CCL21 is important for tumor invasion to lymph nodes, and CXCL12 may explain downregulation of CD26 on the cell surface. CXCL13 expression in lymphoid follicular formation in skin and CCR3 expression on tumor cells in CD30(+) lymphoproliferative disorders are also discussed. Biologics targeting chemokines and their receptors are promising strategies for cutaneous lymphoma. Indeed, humanized anti-CCR4 monoclonal antibody showed potent antitumor activity against CCR4(+) lymphoma cells both in vitro and ex vivo. This antibody may also be useful for allergic diseases such as hay fever. Further study on chemokines and chemokine receptors will be helpful for new classification of cutaneous lymphoma, elucidation of pathogenesis, and development of new therapeutic strategies. PMID- 21628852 TI - [Successful bosentan therapy in a case of pulmonary arterial hypertention developed during immunosuppressive therapy for lupus nephritis]. AB - We report a 43-year-old female who developed pulmonary arterial hypertension (PAH) during intensive immunosuppressive therapy for systematic lupus erythematosus (SLE). She was diagnosed as SLE at the age of 32 years based on serological and hematological abnormalities, oral ulcers, and facial erythema. She experienced frequent flare-ups of disseminated discoid lupus between the ages of 33 and 36 years and developed immune thrombocytopenia at the age of 39 years. In 2007 when she was 43 years old, she developed lupus nephritis (LN) with elevated serum anti-double stranded DNA antibodies and urine protein of less than 1 g/day. Combination therapy for the LN with 35 mg/day prednisolone and intravenous cyclophosphamide (IVCY) led to renal remission. After the seventh monthly session of IVCY, she developed dyspnea on exertion. PAH was diagnosed based on enlarged main pulmonary arteries on the chest x-ray, right ventricular outflow and a peak tricuspid regurgitant pressure gradient exceeding 45 mmHg on echocardiography, an elevated plasma brain natriuretic peptide (BNP) level of 260 pg/ml, the exclusion of pulmonary thromboembolism, and no lung fibrosis. The PAH was treated successfully with bosentan. At present the tricuspid regurgitation has disappeared, and the plasma BNP level has normalized. PMID- 21628853 TI - [Case of systemic-onset juvenile idiopathic arthritis complicated with sepsis - usefulness of cytokine profile for the differentiation between macrophage activation syndrome and sepsis]. AB - The differential diagnosis of macrophage activation syndrome (MAS) and sepsis must be considered in the clinical course of systemic-onset juvenile idiopathic arthritis (s-JIA) with sudden onset of high-grade fever and abnormal laboratory findings, including leukocytopenia, thrombocytopenia, and coagulopathy. In this report, we describe the case of a 17-month-old girl diagnosed with s-JIA complicated with sepsis. Her serum interleukin (IL)-18 level was significantly elevated throughout the clinical course. Furthermore, compared to other MAS patients, she showed a significantly elevated serum IL-6 level and procalcitonin in sepsis. Therefore, our results suggest that a patient's cytokine profile may be a useful indicator of disease activity and may thus help in the differential diagnosis of sepsis and MAS in s-JIA. PMID- 21628854 TI - Importance of relative dose intensity in chemotherapy for diffuse large B-cell lymphoma. AB - CHOP therapy combined with rituximab (R-CHOP) is currently a standard chemotherapy for diffuse large B-cell lymphoma (DLBCL). However, relapse is detected despite R-CHOP in approximately 30% of patients. Treatment results should be further improved. Previously, second- and third-generation therapies such as MACOP-B, m-BACOD, and ProMACE-CytaBOM were performed to improve the results of DLBCL treatment. However, dose intensity (DI) enhancement increased treatment-associated toxicity, and the treatment results did not improve. Recently, the entity of the relative dose intensity (RDI) was proposed as an index of the intensity of chemotherapy. In this method, the ratio of actual DI to the DI designed per specific period is numerically evaluated. The purpose of calculating the RDI is to achieve chemotherapy as scheduled while maintaining the DI, and not to improve the DI. Previous studies reported that the maintenance of the RDI during CHOP therapy improved the treatment results. In this paper, we review DI and RDI in studies of DLBCL, and revisit the significance of these indicators. PMID- 21628855 TI - BCL2 and MYC dual-hit lymphoma/leukemia. AB - Translocation of the BCL2 gene on the chromosome band 18q21.3 results in consistent expression of the Bcl2 protein, an apoptosis inhibitor. BCL2 usually translocates to the immunoglobulin (IG) heavy chain (IGH) gene as t(14;18)(q32;q21.3) and rarely to IG light chain (IGK, IGL) loci as t(2;18)(p11;q21.3) or t(18;22)(q21.3;q11). The t(14;18) translocation is observed in 70-95% of follicular lymphoma cases and 20-30% of diffuse large B-cell lymphoma (DLBCL) cases. The MYC gene on chromosome band 8q24 acts as an accelerator of cell proliferation. MYC translocates to 14q32/IGH as t(8;14)(q24;q32) or less commonly to 2p11/IGK as t(2;8)(p11;q24) or 22q11/IGL as t(8;22)(q24;q11). The 8q24/MYC translocation is detected in nearly all Burkitt lymphoma (BL) and up to 10% of DLBCL cases. Both translocations rarely occur in an identical cell and this lymphoid malignancy is termed BCL2 and MYC dual-hit lymphoma/leukemia (DHL). The pathological diagnosis in most cases of DHL with BCL2-IG and MYC-IG translocation is B-cell lymphoma, unclassifiable, with features intermediate between DLBCL and BL, although DLBCL is most common in DHL with BCL2-IG and MYC-nonIG translocation. The frequency of DHL with BCL2 and MYC translocation is estimated at around 2% of all B-cell malignancies. The condition is characterized by elevated serum lactate dehydrogenase levels, the presence of B symptoms, bone marrow involvement, advanced disease stage, extranodal involvement, and central nervous system (CNS) involvement at presentation or disease progression. Despite treatment strategies including CNS-targeted therapy, the prognosis for DHL is extremely poor. In this review, the current knowledge of the clinicopathological status of DHL is summarized and discussed. PMID- 21628856 TI - IgG4-related disease: a novel lymphoproliferative disorder discovered and established in Japan in the 21st century. AB - IgG4-related disease is a novel lymphoproliferative disorder that shows hyper IgG4-gamma-globulinemia and IgG4-producing plasma cell expansion in affected organs with fibrotic or sclerotic changes. Patients show systemic inflammatory conditions and various symptoms depending on the affected organ. Since the first report of patients with elevated serum IgG4 in sclerosing pancreatitis in 2001, various systemic disorders described by many names have been reported. Despite similarities in the organs involved in IgG4-related Mikulicz's disease and Sjogren's syndrome, there are marked clinical and pathological differences between these conditions. Most patients diagnosed with autoimmune pancreatitis in Japan have IgG4-related pancreatitis [Type 1 autoimmune pancreatitis (AIP), lymphoplasmacytic sclerosing pancreatitis (LPSP)], a disease distinct from some of the western type [Type 2 AIP, idiopathic duct-centric chronic pancreatitis (IDCP), autoimmune pancreatitis with granulocytic epithelial lesions (GEL)]. Diagnosis of IgG4-related disease is characterized by both elevated serum IgG4 (>135 mg/dL) and histopathological features including lymphocyte and IgG4(+) plasma cell infiltration (IgG4(+) plasma cells/IgG(+) plasma cells>40%). Differential diagnosis from other distinct disorders, such as sarcoidosis, Castleman's disease, Wegener's granulomatosis, lymphoma, cancer, and other existing conditions associated with high serum IgG4 level or abundant IgG4 bearing plasma cells in tissues is necessary. We have begun a clinical prospective study to establish a treatment strategy (Phase II prospective treatment study for IgG4-multiorgan lymphoproliferative syndrome: UMIN R000002311). PMID- 21628857 TI - The diagnosis and management of extranodal NK/T-cell lymphoma, nasal-type and aggressive NK-cell leukemia. AB - Natural killer (NK) cell lymphomas are rare malignancies. They are classified as extranodal NK/T-cell lymphoma, nasal type, and aggressive NK cell leukemia. NK cell neoplasms are prevalent in Asian and South American populations, but are extremely rare in the West. They can be classified clinically into nasal, non nasal, and aggressive lymphoma/leukemia subtypes. For nasal NK cell lymphomas, combined chemotherapy and radiotherapy are indicated for stage I/II disease. Chemotherapy is the main treatment for stage III/IV nasal NK cell lymphomas, as well as the non-nasal and aggressive subtypes. Regimens containing drugs not affected by the P-glycoprotein, particularly in combination with L-asparaginase, have resulted in much improvement in treatment outcome for high-risk, refractory or relapsed patients. Autologous or allogeneic hematopoietic stem cell transplantation should be considered for selected patients. Epstein-Barr virus DNA load as a surrogate marker for prognostication, and clinical stratification of patients should be incorporated in clinical management algorithms. PMID- 21628858 TI - Comparison of long-term clinical outcomes of CHOP chemotherapy between Japanese patients with nodal peripheral T-cell lymphomas and those with diffuse large B cell lymphoma in the study group of the Tohoku Hematology Forum. AB - To clarify the clinical outcome of peripheral T-cell lymphomas (PTCLs), we conducted a retrospective review comparing the outcomes of patients with PTCL (nodal peripheral T-cell lymphoma, unspecified, n=34 ; angioimmunoblastic T-cell lymphoma, n=12) to those with diffuse large B-cell lymphoma (DLBCL, n=48). All patients received CHOP-based chemotherapy without rituximab. PTCL patients presented at a more advanced clinical stage (91% vs. 65%, P<0.002) with a poorer performance status (26% vs. 17%, P<0.002) than DLBCL patients. The complete response rate among PTCL patients was significantly lower than among DLBCL patients (39% vs. 67%, P<0.008), as was the 3-year overall survival rate (26% vs. 50%, P=0.005), and Cox multivariate analysis revealed immunophenotype, performance status, and extranodal site involved to be significantly associated with shorter overall survival (P=0.045, P=0.007, and P=0.034, respectively). Our findings suggest that PTCL patients tend to have a poor prognosis associated with several initial risk factors. Moreover, the T-cell phenotype itself appears to have a significant impact on overall survival. Thus, standard CHOP chemotherapy may be inadequate for PTCLs, especially in patients with high-risk factors. The development of newly stratified therapies for the treatment of PTCLs would be highly desirable. PMID- 21628859 TI - A highly therapy-resistant case of B-cell lymphoma, unclassifiable, with features intermediate between diffuse large B-cell lymphoma and Burkitt lymphoma showing strong BCL2 staining, otherwise indistinguishable from Burkitt lymphoma. AB - B-cell lymphoma, unclassifiable, with features intermediate between diffuse large B-cell lymphoma and Burkitt lymphoma (iDLBCL/BL) is a newly categorized lymphoma in the WHO Classification of Tumors of Haematopoietic and Lymphoid Tissues v4. This lymphoma has characteristics of both DLBCL and BL, morphologically and genetically. We report a case with therapy-resistant iDLBCL/BL, which used to be categorized as BL in the WHO Classification v3. This case showed strong BCL2 staining, otherwise exhibiting features of BL. The case was treated with intensive chemotherapy containing high-dose methotrexate, cyclophosphamide, and cytarabine. However, soon after the chemotherapy, the lymphoma relapsed in the central nervous system and was resistant to whole-brain radiation therapy and allogeneic stem cell transplantation. The patient died 37 days after the transplantation. The clinical course was different from that of typical BL in terms of resistance to intensive chemotherapy, in agreement with the characteristics of iDLBCL/BL. PMID- 21628860 TI - Double-hit lymphoma at second relapse of Burkitt-like lymphoma: a case report. AB - Double-hit lymphoma (DHL) is a rare and extremely unfavorable type of lymphoma with concurrent chromosomal translocations of BCL2 and MYC. It is considered that BCL2 translocation precedes MYC events in lymphomagenesis of DHL. In fact, most cases of DHL arise de novo or following FL. We describe a very rare case of DHL arising from Burkitt-like lymphoma according to the revised European-American classification of lymphoid neoplasms. A 67-year-old Japanese male presented with persistent fever. [(18)F]-fluorodeoxyglucose positron emission tomography revealed multiple abnormal accumulations in the bone marrow, pancreas, and periphery of the left kidney. The patient was diagnosed with Burkitt-like lymphoma according to a bone marrow biopsy. At the disease onset and the first relapse, chemotherapy was effective and the patient experienced sustained and complete remission. At the second relapse, however, the clinical presentation and morphology of lymphoma cells were nearly identical, but a high level of chemoresistance was acquired, and the patient succumbed almost 1 month after hospitalization. Chromosomal analyses revealed a complex karyotype with concurrent t(14;18) and t(8;22) translocations, which have not been previously detected. It is therefore important to note that DHL cannot be diagnosed without chromosomal analysis. Cytogenetic analyses should thus be performed for patients with high-grade B-cell lymphoma and who experience a recurrence of this lymphoma. PMID- 21628861 TI - Bilateral conjunctival lesions in blastic plasmacytoid dendritic cell neoplasm. AB - The purpose of this study is to report on a patient who developed conjunctival lesions of blastic plasmacytoid dendritic cell neoplasm (BPDCN) after radiation to skin lesions of the same pathological type. A 79-year-old man developed salmon pink lesions in the lower fornix of the conjunctiva of both eyes and biopsy revealed BPDCN. One and a half years previously, he noticed an erythematous plaque with a 30-mm diameter, which later became multiple, on the left chest, and the biopsy revealed BPDCN. The bone marrow was negative for CD56-positive cells, but fluorescence-activated cell sorting analysis of peripheral blood cells revealed a group of cells positive for CD4 and CD56, or CD4 and CD123. The monocyte fraction, in an increased percentage of white blood cell counts, did contain atypical cells positive for the three markers. Whole-body 2-[(18)F]fluoro 2-deoxy-D-glucose positron emission tomography combined with computed tomography demonstrated no abnormal uptake lesions. He received 30 Gy of radiation to the chest lesions, and later, the same dose of radiation to novel skin lesions on the back, waist, and chest, and also to pharyngeal lesions. In conclusion, the conjunctiva could be involved with BPDCN and pathological differential diagnosis by biopsy is mandatory to establish the correct diagnosis. PMID- 21628862 TI - Unrelated bone marrow transplantation induced long-term remission in a patient with life-threatening Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis. AB - We present a case of life-threatening Epstein-Barr virus (EBV)-associated hemophagocytic lymphohistiocytosis (HLH) with severe hepatitis that was successfully treated by allogeneic stem cell transplantation from an unrelated donor. A 26-year-old woman was admitted to hospital with a high fever and liver dysfunction. Laboratory tests, including bone marrow aspiration, revealed severe HLH that occurred after EBV infection. High-dose methylprednisolone and etoposide therapy did not control the disease. We could control the HLH, but the EBV viremia continued following the CHOPE (cyclophosphamide, doxorubicin, vincristine, prednisone, and etoposide) chemotherapy regimen. Therefore, the patient underwent allogeneic bone marrow transplantation from an HLA-matched, unrelated donor. The patient has remained in good condition without disease recurrence for 2 years after bone marrow transplantation. Although there is no consensus regarding allogeneic stem cell transplantation for EBV-HLH, it is the treatment of choice for aggressive EBV-HLH when the patient is refractory to intensive chemotherapy. PMID- 21628863 TI - Virus-associated hemophagocytic syndrome caused by pandemic swine-origin influenza A (H1N1) in a patient after unrelated bone marrow transplantation. PMID- 21628864 TI - Differentiation of uterine natural killer cells in pregnant SCID (scid/scid) mice. AB - To determine whether functional T- and B-cells can affect differentiation and/or proliferation of uterine natural killer (uNK) cells, their numbers in SCID mice (genotype, C.B.-17/Icr-scid/scid) were compared with those of control mice (genotype, C.B.-17/Icr-+/+) on days 8, 12 and 16 of pregnancy. Using biotinylated Dolichos biflorus agglutinin (DBA) lectin staining, uNK cells can be readily classified into 4 subtypes, I to IV, from immature to mature types. The number of uNK cells was significantly lower in the decidua basalis of SCID mice than in that of control mice on day 8 of pregnancy. Particularly, the number of uNK cells of immature subtype II was significantly lower in SCID mice than in the control mice. By day 12, however, the uNK cell number in the SCID mice reached the same level as that of the control mice. It is likely that uNK cell differentiation in SCID mice was delayed during the early placentation period due to a lack of functional T and B cells. PMID- 21628865 TI - Immunohistochemical observation of canine degenerative myelopathy in two Pembroke Welsh Corgi dogs. AB - Immunohistochemistry was performed to assess whether oxidative stress and/or denatured proteins play roles in the pathogenesis of canine degenerative myelopathy (DM). Two Pembroke Welsh Corgi (PWC) dogs with a homozygous mutation (c.118G>A) in the canine superoxide dismutase 1 (SOD1) gene were examined. The pathological features of the dogs were consistent with those of previous cases of DM in PWC. In the spinal lesions, diffuse SOD1 expression was observed in the neurons while no inclusion-like aggregates had formed, which disagreed with the findings of a previous study. A unique inducible nitric oxide synthase (iNOS) staining pattern in reactive astrocytes and a significant increase in ubiquitin immunoreactivity in the spinal lesions were also observed. These findings indicate the involvement of oxidative stress and the accumulation of ubiquitinated proteins in the pathogenesis of canine DM, whereas the role of SOD1 remains unclear. PMID- 21628866 TI - Immune responses to new vaccine candidates constructed by a live attenuated Salmonella Typhimurium delivery system expressing Escherichia coli F4, F5, F6, F41 and intimin adhesin antigens in a murine model. AB - To construct a novel live oral vaccine candidate for the prevention of pathogenic Escherichia coli infections in neonatal piglets, an expression and secretion plasmid and an attenuated Salmonella delivery system were used. The individual E. coli genes K88ac, K99, FasA, F41 and intimin adhesins were inserted into pBP244 containing asd, lepB, secA and secB, and these plasmids were transformed into a Salmonella Typhimurium DeltacpxR Deltalon Deltaasd. Forty female BALB/c mice were divided into four groups, A to D (ten mice per group). Groups A and B were administered with the mixture containing all constructs and the S. Typhimurium containing pBP244 only as a control, respectively. Groups C and D were primed and boosted with the mixture and the S. Typhimurium harboring pBP244 only, respectively. Each recombinant adhesin secreted from the individual candidates was confirmed by Western blot analysis. The serum IgG and secretory IgA (sIgA) titers to individual adhesins in all immunized groups were higher than those in control. Furthermore, IgG and sIgA levels in group C were higher than those in group A, and the IgG1 titers were increased in Group C but IgG2a titers were similar or decreased in Group C compared to Group A. In addition, the vaccine strains were not detected in fecal samples of any immunized mice. The novel vaccine candidates are not only highly immunogenic, but also safe for vaccinated mice and environment. In addition, the immune responses can be more efficiently induced through the booster-administration. PMID- 21628867 TI - Astrocytoma in an African hedgehog (Atelerix albiventris) suspected wobbly hedgehog syndrome. AB - A 28-month-old African hedgehog was referred to our hospital with progressive tetraparesis. On the first presentation, the hedgehog was suspected as having wobbly hedgehog syndrome (WHS) and the animal was treated with medication and rehabilitation. The animal died 22 days after onset. Pathological examination revealed that the animal was involved in astrocytoma between the medulla oblongata and the spinal cord (C1). This report indicates that a primary central nervous system tumor should be considered as one of the differential diagnoses for hedgehogs presenting with progressive paresis, together with WHS. PMID- 21628868 TI - Haplotype-based association of four lymphotoxin-alpha gene polymorphisms with the risk of coronary artery disease in Han Chinese. AB - Lymphotoxin-alpha (LTA), a pro-inflammatory cytokine, has been implicated in the pathogenesis of coronary atherosclerosis. Meanwhile, association of some single nucleotide polymorphisms (SNPs) of LTA gene with coronary artery disease (CAD) has been evaluated; however, the results are irreproducible. We therefore investigated the relationship between four SNPs of LTA gene and CAD in Han Chinese: G+10A (rs1800683, 5'-untranslated region), A+80C (rs2239704, 5' untranslated region), T+496C (Cys13Arg, rs2229094, exon 2), and C+804A (Thr26Asn, rs1041981, exon 3). Genotyping was performed in 438 CAD patients and 330 healthy controls. Single-locus analysis showed that the genotype and allele frequencies of G+10A polymorphism exhibited marginal differences between CAD patients and controls, although no statistical significance was observed after the Bonferroni correction. Logistic regression analysis revealed that GG genotype of G+10A polymorphism was significantly associated with the risk of CAD under the dominant mode, whereas no significant association was detected between A+80C polymorphism and CAD. In contrast, individuals carrying TT or TC genotype of T+496C polymorphism showed a decreased CAD risk relative to those with CC genotype under the recessive mode. Likewise, CC genotype of C+804A polymorphism was associated with a protective effect on CAD under the dominant mode. Further, in haplotype analysis, the haplotype G-C-T-C (in order of rs1800683, rs2239704, rs2229094 and rs1041981) was significantly associated with a decreased risk of CAD after assigning the most common haplotype A-C-T-A as a reference. In conclusion, we show a protective effect of the haplotype G-C-T-C on the occurrence of CAD, suggesting the involvement of LTA in CAD pathogenesis. PMID- 21628869 TI - C5a, a complement activation product, is a useful marker in predicting the severity of necrotizing enterocolitis. AB - Necrotizing enterocolitis (NEC) is the most common neonatal gastrointestinal emergency, predominantly affecting low-birth weight, premature infants. Early clinical signs of NEC are nonspecific and the laboratory findings are not fully reliable. Its severe morbidities and rapid progression require the application of new biomarkers for early diagnosis and intervention. The complement activation product, C5a (anaphylatoxin) has been reported to be a contributing factor leading to mesenteric ischemia/reperfusion injury which is a predisposing factor in the pathogenesis of NEC. Therefore, our aim was to evaluate the efficacy of serial C5a measurements in the diagnosis and follow-up of NEC. Preterm infants, whose gestational age and weight matched each other, were grouped as controls (n = 23) and NEC (n = 22). Serum levels of C5a, serum amyloid-A (SAA), C-reactive protein (CRP), and interleukin-6 (IL-6) levels were measured on the third day of life for the control group and on the day of diagnosis (1st day), 3rd and 7th days of the NEC group. C5a, SSA, CRP, and IL-6 levels were significantly higher in the NEC patients compared to the control group (P < 0.05) in the follow-up. Additionally, serum levels of C5a were found to be more accurate than the other parameters for the prediction of death and requirement for surgery at the time of diagnosis (P < 0.05). In conclusion, C5a may be useful as a new marker for both diagnosis and follow-up of infants with NEC in combination with clinical and radiographical findings. PMID- 21628870 TI - Clinicopathological features of malignant lymphoma in Japan: the Miyagi Study. AB - The Miyagi Study is an epidemiological study of malignant lymphoma, including immunological and genetic analyses, constructed by a population-based registration system covering Miyagi prefecture, Japan. A total of 1,552 newly diagnosed cases in Miyagi between 2002 and 2008 were enrolled in this study; 75% were B-cell lymphomas, 19% were T-cell and natural killer-cell (T/NK-cell) lymphomas, and 5% were Hodgkin's lymphomas. The most frequent subtype of B-cell lymphoma is diffuse large B-cell lymphoma, followed by follicular lymphoma and extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (51%, 24% and 8%, respectively). Thus, follicular lymphoma accounts for 18.2% of newly diagnosed cases in Miyagi; unexpectedly, its frequency is similar to that reported in Western countries. The common subtypes of T/NK-cell lymphoma are peripheral T-cell lymphoma, angioimmunoblastic T-cell lymphoma, and adult T-cell leukemia/lymphoma (30%, 15% and 14%, respectively). Most of the data are similar to those reported in Asian countries, except for follicular lymphoma. We also analyzed the CD20 expression in B-cell lymphomas by flow cytometry for the cell membrane expression and by immunohistochemistry for the cytoplasmic expression. The cell membrane expression of CD20 protein may determine the susceptibility of B-cell lymphomas to anti-CD20 antibody therapy. The lack of CD20 expression was confirmed by both methods in 4 cases of 585 newly diagnosed cases (0.7%) and in 5 of 67 recurrent cases (7.5%). Furthermore, 23 cases (6.5%) showed the discrepancy of CD20 expression between both methods. The Miyagi Study has revealed the latest epidemiological features of malignant lymphoma in Japan. PMID- 21628871 TI - Cellular physiology of channels and transporters in gastrointestinal tracts. Foreword. PMID- 21628872 TI - The glycine transporter GLYT1 in human intestine: expression and function. AB - Glycine is a well-documented cytoprotective agent and protects mammalian intestine against ischemia-reperfusion injury, irradiation and experimentally induced colitis. The specific glycine transporter GLYT1 is found throughout the human intestine where it is responsible for some 30-50% of glycine uptake into intestinal epithelial cells across the basolateral membrane and appears to function to maintain glycine supply to enterocytes and colonocytes. This paper reviews current knowledge of GLYT1 and presents recent evidence supporting its essential role in glycine mediated cytoprotection in intestinal absorptive cells. Regulatory mechanisms involved in intestinal expression of GLYT1 are discussed and the potential of glycine for use as an anti-inflammatory, protective agent in the management of inflammatory bowel disease examined. PMID- 21628873 TI - Regulation of colonic ion transport by gasotransmitters. AB - Gaseous molecules such as nitric oxide (NO), hydrogen sulfide (H2S), or carbon monoxide (CO) are involved in the regulation of colonic water and salt transport, which can be switched between absorption and secretion. Nitric oxide is produced from the amino acid L-arginine by different isoforms of the enzyme NO synthase, which are expressed both by enteric neurones and by the colonic epithelium. NO donors evoke a transepithelial Cl- secretion in vitro. Most actions of NO are mediated by a stimulation of guanosine 5' cyclic monophosphate (cGMP) synthesis via activation of the soluble guanylate cyclase. In rat colon, NO possesses several main action sites: a stimulation of apical Cl- channels most probably not related to cGMP-dependent phosphorylation, and an increase in the cytosolic Ca2+ concentration, which stimulates a Ca2+-dependent K+ conductance in the basolateral membrane. Hydrogen sulfide, produced during the metabolism of the amino acid L-cysteine, also evokes a Cl- secretion, either by stimulation of secretomotor submucosal neurones as in guinea-pig colon or by activating Ca2+ dependent and ATP-sensitive K+ channels as in rat colon. The third gasotransmitter, CO, produced during the degradation of heme, evokes anion secretion carried by Cl- and HCO3-. This response is mainly caused by the activation of apical anion channels and a stimulation of Ca2+-dependent K+ channels via an increase of the cytosolic Ca2+ concentration. Consequently, gaseous molecules produced by enteric neurones, epithelial cells, as well-in the case of H2S-the microbial flora affect key transport enzymes involved in colonic ion transport. PMID- 21628874 TI - News from the end of the gut--how the highly segmental pattern of colonic HCO3- transport relates to absorptive function and mucosal integrity. AB - A number of transport mechanisms in the colonic epithelium contribute to HCO3- movement across the apical and basolateral membranes, but this ion has been largely regarded as a by-product of the transport functions it is involved in, such as NaCl or short chain fatty acid (SCFA) absorption. However, emerging data points to several specific roles of HCO3- for colonic epithelial physiology, including pH control in the colonic surface microenvironment, which is important for transport and immune functions, as well as the secretion and the rheological properties of the mucus gel. Furthermore, recent studies have demonstrated that colonic HCO3- transporters are expressed in a highly segmental as well as species specific manner. This review summarizes recently gathered information on the functional anatomy of the colon, the roles of HCO3- in the colonic epithelium, colonic mucosal integrity, and the expression and function of HCO3- transporting mechanisms in health and disease. PMID- 21628875 TI - Extracellular pH in restricted domains as a gating signal for ion channels involved in transepithelial transport. AB - The importance of intracellular pH (pH(i)) in the regulation of diverse cellular activities ranging from cell proliferation and differentiation to cell cycle, migration and apoptosis has long been recognised. More recently, extracellular pH (pH0), in particular that of relatively inaccessible compartments or domains that occur between cells in tissues, has begun to be acknowledged as a relevant signal in cell regulation. This should not be surprising given the abundant reports highlighting the pH0-dependence of the activity of membrane proteins facing the extracellular space such as receptors, transporters, ion channels and enzymes. Changes in pH affect the ionisation state of proteins through the effect on their titratable groups. There are proteins, however, which respond to pH shifts with conformational changes that are crucial for catalysis or transport activity. In such cases protons act as signalling molecules capable of eliciting fast and localised responses. We provide examples of ion channels that appear fastidiously designed to respond to extracellular pH in a manner that suggests specific functions in transporting epithelia. We shall also present ideas as to how these channels participate in complex transepithelial transport processes and provide preliminary experiments illustrating a new way to gauge pH0 in confined spaces of native epithelial tissue. PMID- 21628876 TI - Function of K+-Cl- cotransporters in the acid secretory mechanism of gastric parietal cells. AB - Gastric proton pump (H+, K+-ATPase) secretes H+ of acid (HCl) via the luminal membrane of parietal cells. For the HCl secretion, Cl-- and K+-transporting proteins are required. Recent our studies have demonstrated that K+-Cl- cotransporters (KCC3a and KCC4) are expressed in gastric parietal cells. KCC3a is associated with Na+, K+-ATPase in the basolateral membrane, and KCC4 is associated with H+, K+-ATPase in the apical canalicular membrane. This paper summarizes the functional association between KCCs and P-type ATPases and the contribution of these complexes to acid secretion in gastric parietal cells. PMID- 21628877 TI - Functional transformation of gastric parietal cells and intracellular trafficking of ion channels/transporters in the apical canalicular membrane associated with acid secretion. AB - The parietal cell of the gastric gland is a highly differentiated cell responsible for the gastric hydrochloric acid secretion into the lumen of the stomach. In response to stimulation of acid secretion, the parietal cells undergo well-characterized morphological transformations to recruit H+/K+-ATPase from the cytoplasmic tubulovesicles to the apical canalicular membrane. Besides H+ extrusion via H+/K+-ATPase, Cl- efflux and K+ recycling across the apical canalicular membrane are necessary via chloride and potassium channels/transporters, respectively. In the last decade, a number of molecular candidates for the Cl- efflux and K+ recycling have been identified in the apical canalicular membrane of the parietal cell. This review focuses on the functional transformation of gastric parietal cells and intracellular trafficking of ion channels/transporters expressed in the apical canalicular membrane associated with gastric acid secretion. PMID- 21628878 TI - Inactivation of cystein-aspartic acid protease (caspase)-1 by saikosaponin A. AB - This work investigates the anti-inflammatory mechanism of saikosaponin A (SA), a major component of Bupleurum falcatum LINNE. SA significantly inhibited phorbol myristate acetate (PMA) plus A23187-induced the production and expression of interleukin (IL)-6 and tumor necrosis factor (TNF)-alpha in human mast cell (HMC) 1 cells. SA suppressed PMA plus A23187-induced phosphorylation of extracellular signal-regulated kinase and p38. When HMC-1 cells were treated with SA, translocation of nuclear factor (NF)-kappaB/Rel A into nucleus and degradation of inhibitor of NF-kappaB (IkappaB) in cytoplasm were inhibited. SA decreased PMA plus A23187-induced cysteine-aspartic acid protease (caspase)-1 activity. IL 1beta production was also inhibited by SA. Finally, SA significantly decreased the number of nasal rubs and serum TNF-alpha level in the ovalbumin-sensitized allergic rhinitis mouse model. The underlying mechanism involves, at least in part, inactivation of caspase-1, which provides new evidence for therapeutic application of SA to target inflammatory processes. PMID- 21628879 TI - Roles of coagulation pathway and factor Xa in the progression of diabetic nephropathy in db/db mice. AB - The active type of coagulation factor X (factor Xa) activates various cell-types through protease-activated receptor 2 (PAR2). We previously reported that a factor Xa inhibitor could suppress Thy-1 nephritis. Considering that fibrin deposition is observed in diabetic nephropathy as well as in glomerulonephritis, this study examined the roles of the coagulation pathway and factor Xa in the development of diabetic nephropathy using type 2 diabetic model mice. Diabetic (db/db) and normoglycemic (m+/m+) mice were immunohistochemically evaluated for their expression/deposition of PAR2, transforming growth factor (TGF)-beta, fibrin, extracellular matrix (ECM) proteins, and CD31 at week 20. Significantly greater numbers of PAR2-positive cells and larger amounts of fibronectin, and collagen IV depositions were observed in the glomeruli of db/db mice than those in m+/m+ mice. Next, expression of PAR2 versus deposition of collagen IV and fibronectin was compared between week 20 and week 30, and the number of PAR2 positive cells in the glomeruli decreased in contrast with the increased accumulation of ECM proteins. In an intervention study, fondaparinux, a factor Xa inhibitor, was subcutaneously administered for ten weeks from week 10 to 20. Fondaparinux treatment significantly suppressed urinary protein, glomerular hypertrophy, fibrin deposition, expression of connective tissue growth factor, and ECM proteins deposition together with CD31-positive capillaries. These results suggest that coagulation pathway and glomerular PAR2 expression are upregulated in the early phase of diabetes, together with the increase of profibrotic cytokines expression, ECM proteins deposition and CD-31-positive vessels. Factor Xa inhibition may ameliorate glomerular neoangiogenesis and ECM accumulation in diabetic nephropathy. PMID- 21628880 TI - Natural plant extract tubeimoside I promotes apoptosis-mediated cell death in cultured human hepatoma (HepG2) cells. AB - Tubeimoside I (TBMS I), an extract from Chinese herbal medicine Bolbostemma paniculatum (MAXIM.) FRANQUET (Cucurbitaceae) has been shown as a potent anti tumor agent for a variety of human cancers, but yet to be evaluated for hepatoma that is highly prevalent in Eastern Asian countries including China. Here, we examined in vitro the cytotoxic effects of TBMS I on human hepatoma (HepG2) and normal liver (L-02) cell lines. We also investigated TBMS I-induced molecular events related to apoptosis in HepG2 cells. The results show that TBMS I inhibited the proliferation of both HepG2 and L-02 cells in a dose- and time dependent manner, but HepG2 cells appeared more sensitive to the agent. When exposed to TBMS I for 24, 48 and 72 h, IC50 for HepG2 cells versus L-02 cells were 15.5 vs. 23.1, 11.7 vs. 16.2, 9.2 vs. 13.1 (uM, p<0.01), respectively. TBMS I induced cell shrinkage, nuclear condensation and fragmentation, cell cycle arrest at the G2/M phase, mitochondrial membrane disruption, release of cytochrome c from the mitochondria, activation of caspase 3 and 9, and shifting Bax/Bcl-2 ratio from being anti-apoptotic to pro-apoptotic, all indicative of initiation and progression of apoptosis involving mitochondrial dysfunction. Taken together, these results indicate for the first time that TBMS I potently inhibited growth in HepG2 cells by mediating a cascade of apoptosis signaling pathways. Considering its sensitivity of HepG2 cells, preferential distribution in the liver and natural product origin, TBMS I therefore may have a great potential as a chemotherapeutic drug candidate for hepatoma. PMID- 21628881 TI - Astaxanthin induces mitochondria-mediated apoptosis in rat hepatocellular carcinoma CBRH-7919 cells. AB - We designed to study the role of mitochondria in astaxanthin-induced apoptosis in hepatocellular carcinoma cells. Effect of astaxanthin on cell proliferation was studied by using methyl thiazolyl tetrazolium (MTT) in three tumor cell lines (CBRH-7919, SHZ-88 and Lewis) and normal human hepatocyte HL-7702 cell. Cell apoptosis rate, changes of mitochondrial morphology, mitochondrial transmembrane potential and electron transport chain were evaluated respectively. Expressions of B cell lymphoma/leukemia-2 (Bcl-2) and Bcl-2 associated X protein (Bax) were detected by Western blot. Results as following, astaxanthin had little effect on HL-7702 cell, however its inhibition was most pronounced in CBRH-7919 cell line with an IC50 of 39 uM. This dose of astaxanthin and CBRH-7919 cell line were chosen for further studies. Astaxanthin could induce cell apoptosis and mitochondrial membrane damage. The mitochondrial transmembrane potential and function of electron transport chain were decreased. The expression of Bcl-2 protein was down-regulated but that of Bax protein was up-regulated. In conclusion, astaxanthin showed anticancer effect by inducing cell apoptosis through the regulation of mitochondrial-dependent manner. PMID- 21628882 TI - Modification and translocation of Rac/Rop guanosine 5'-triphosphate-binding proteins of Scoparia dulcis in response to stimulation with methyl jasmonate. AB - Translocation of two Rac/Rop guanosine 5'-triphosphate-binding proteins from Scoparia dulcis, Sdrac-1 and Sdrac-2, was examined employing transformed belladonna which overproduces these proteins as glutathione-S-transferase-tagged forms. The transferase activities of the fused proteins in microsomal fraction of belladonna markedly increased by the incubation with methyl jasmonate either in Sdrac-1 or Sdrac-2 transformant, while low and constant activities were observed in the untreated control. Recombinant Sdrac-2 protein was found to bind to prenyl chain in the presence of cell extracts prepared from methyl jasmonate-treated S. dulcis, however, Sdrac-1 was palmitoylated by the addition of the cell extracts. These results suggest that both Sdrac-1 and Sdrac-2 translocate to plant membranes by the stimulation with methyl jasmonate, however, targeting of these proteins is triggered by the independent modification mechanisms, palmitoylation for Sdrac-1 and prenylation for Sdrac-2. PMID- 21628883 TI - Rhapontigenin inhibited hypoxia inducible factor 1 alpha accumulation and angiogenesis in hypoxic PC-3 prostate cancer cells. AB - Hypoxia inducible factor 1 alpha (HIF-1alpha) is frequently over-expressed in the numerous types of cancer and plays an important role in angiogenesis. In the present study, the inhibitory mechanism of rhapontigenin isolated from Vitis coignetiae was investigated on HIF-1alpha stability and angiogenesis in human prostate cancer PC-3 cells. Rhapontigenin significantly suppressed HIF-1alpha accumulation at protein level but not at mRNA level in PC-3 cells under hypoxia. Also, rhapontigenin suppressed hypoxia-induced HIF-1alpha activation in various cancer cells, such as colorectal adenocarcinoma (SW620), breast adenocarcinoma (MCF-7), fibrosarcoma (HT-1080) and prostate carcinoma (LNCaP). Interestingly, rhapontigenin had more potency in inhibition of hypoxia-induced HIF-1alpha expression than that of resveratrol, a known HIF-1alpha inhibitor. In addition, rhapontigenin promoted hypoxia-induced HIF-1alpha degradation and cycloheximide (CHX) blocked protein synthesis. A prolyl hydroxylase (PHD) inhibitor dimethyloxalylglycine (DMOG) is usually utilized to examine whether prolyl hydroxylation is involved in inhibition of HIF-1alpha accumulation. Here, DMOG recovered HIF-1alpha accumulation inhibited by rhapontigenin. Immunoprecipitation assay also revealed that rhapotigenin enhanced the binding of hydroxylated HIF 1alpha to von Hippel-Lindau (VHL) tumor suppressor protein. Furthermore, rhapontigenin reduced vascular endothelial growth factor (VEGF) secretion in hypoxic PC-3 cells as well as suppressed tube formation in human umbilical vein endothelial cells (HUVECs) treated by the conditioned media of hypoxic PC-3 cells. However, anti-angiogenic effect of rhapontigenin in hypoxic PC-3 cells was reversed by DMOG. Taken together, these findings suggest that rhapontigenin inhibits HIF-1alpha accumulation and angiogenesis in PC-3 prostate cancer cells. PMID- 21628884 TI - Effects of perfluorinated fatty acids with different carbon chain length on fatty acid profiles of hepatic lipids in mice. AB - Alterations by perfluorinated fatty acids (PFCAs) with a chain length of 6-9 carbons in the fatty acid profile of hepatic lipids of mice were investigated. The characteristic changes caused by all the PFCAs examined were increases in the contents and proportions of oleic acid (18 : 1), palmitoleic acid (16 : 1) and 8,11,14-eicosatrienoic acid (20 : 3) in hepatic lipids. Hepatic contents of palmitic acid were also increased by the treatments with the PFCAs. These effects were almost dependent on the hepatic concentrations of PFCA molecules regardless of their carbon chain length. Perfluorooctanoic acid elevated the expressions of mRNA encoding acetyl-CoA carboxylase, fatty acid synthase, malic enzyme, stearoyl CoA desaturase (SCD) (SCD1 and 2), chain elongase (ELOVL5), Delta6 desaturase (Fads2), 1-acylglycerophosphocholine acyltransferase (LPCAT) (LPCAT3). The four PFCAs examined induced microsomal SCD and LPCAT in hepatic concentration dependent manners regardless of carbon chain length. One linear regression line was confirmed between LPCAT activity and hepatic concentration of PFCA at wide range of the concentration, whereas the induction of SCD was saturable at relatively low concentration of PFCAs. These results suggest (i) that PFCAs with a chain length of 6-9 carbons change the fatty acid profile of hepatic lipids by increasing contents and proportions of 16 : 1, 18 : 1 and 20 : 3, (ii) that these alterations in fatty acid profile are caused by up-regulation of SCD, de novo fatty acid synthesis, chain elongase and Delta6 desaturase and (iii) that the mechanism underlying SCD induction is, in part, mediated through peroxisome proliferator-activated receptor alpha. PMID- 21628885 TI - Quantitative comparison of adipocytokine gene expression during adipocyte maturation in non-obese and obese rats. AB - Adipocytokines secreted from adipocytes have been extensively analyzed due to their role as key factors in various complications of obesity, including arterial sclerosis, liver steatosis, insulin resistance, and diabetes. Several in vivo and in vitro studies have suggested that adipocyte maturation is related to fluctuations in adipocytokine secretion. However, the relationship between adipocyte maturation and adipocytokine levels has not been fully elucidated. Therefore, we sought to clarify the link between adipocytokine gene expression and adipocyte maturation through systematic analysis. We quantified mRNA for six adipocytokine genes: adiponectin, resistin, leptin, plasminogen activator inhibitor 1 (PAI-1), heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF), and visfatin, in adipose tissue, in primary cultured adipocytes obtained from an obese Zucker rat, and in the preadipocyte cell line 3T3-L1. Moreover, to elucidate the role of adipocytokines in adipocyte maturation, adipocytokine expression levels were analyzed during maturation. Although fluctuations in adipocytokine gene expression were heterogeneous, gene expression was highly similar during maturation of primary cultured adipocytes from obese and non-obese rats, suggesting that the maturation process is independent from processes that lead to obesity. Moreover, the expression patterns of adiponectin, resistin and leptin mRNA in 3T3-L1 cells were highly similar to those in primary cultured adipocytes, indicating that these adipocytokines could be common maturation markers for primary cultured adipocytes obtained from obese and non obese rats, and for preadipocyte cell lines. PMID- 21628886 TI - Metabonomics study of brain-specific human S100B transgenic mice by using high performance liquid chromatography coupled with quadrupole time of flight mass spectrometry. AB - For the purpose of investigating the effects of S100B on the development of Parkinsion's disease (PD), a high-performance liquid chromatography coupled with electrospray ionization time-of-flight mass spectrometry (HPLC/MS-ESI-TOF) metabonomic approach was established to study the mesencephalon profiling of brain-specific human S100B transgenic mice. In order to obtain more full-scale chemical information of metabolites, two kinds of separation mechanism, including reversed-phase (RP) column chromatography and hydrophilic interaction liquid chromatography (HILIC) column, were combined to use. Acquired data were subjected to principal component analysis (PCA) to investigate the effects of S100B protein on mice mesencephalon metabolite profiles. Potential biomarkers were screened by using Mass Hunter Prossional Profiller (MPP) and were identified by the accurate mass. Twelve metabolites in mesencephalon of S100B transgenic mice were identified as potential biomarkers, among which, glutamic acid (Glu) detected by RP/MS in negative ionization mode, gamma-aminobutyric acid (GABA) and tryptophan (Trp) detected by HILIC/MS in positive ionization mode, phenylalanine (Phe) and histidine (His) detected by HILIC/MS in negative ionization mode, related to metabolic pathway of neurotransmitters in mice central nervous system. The analytical technique used in this paper was able to detect biochemical changes in mesencephalon of S100B transgenic mice, which may be helpful to understand the action mechanism of S100B protein in the development of PD. PMID- 21628887 TI - Development and validation of an LC/MS/MS method for the determination of tenofovir in monkey plasma. AB - A simple, specific, sensitive LC/MS/MS method for the quantitation of tenofovir (TFV) in monkey plasma was developed and validated. After the addition of adefovir as an internal standard (IS), methanol was used to produce a protein free extract. Isocratic chromatographic separation was performed on a reverse phase Discovery C(18) column (4.6*250 mm, 5 um). The mobile phase consisted of methanol-water-formic acid (20 : 80 : 0.5, v/v/v). Detection of TFV and the IS was achieved with electrospray ionization (ESI)-MS/MS in the positive ion mode using 288/176 and 274/162 transitions, respectively. The analytical range was set at 0.005-1.250 ug/ml using a 200 ul plasma sample. The intra- and inter-day precision values were less than 11.4%, and accuracy ranged from 0.4 to 2.9% in all quality control samples. The method was fully validated for its sensitivity, selectivity, accuracy and precision, matrix effect, recovery, and stability. Due to the high polarity of TFV, the major challenge was to circumvent ion suppression when quantitating the plasma concentration of TFV using the LC/MS/MS method. To avoid ion suppression, sufficient chromatographic separation was the most effective means for the present purposes. Moreover, it was found that the reconstitution solvents of the dried residue had a significant impact on LC peak shapes. The validated method was successfully applied to a bioequivalence study in 6 monkeys after the oral administration of two ester prodrugs of TFV (equivalent to TFV 20 mg/kg). The method permits laboratory scientists with access to the appropriate instrumentation to perform rapid TFV determination. PMID- 21628888 TI - Identification of a novel carbohydrate-mimicking octapeptide from chemical peptide library and characterization as selectin inhibitor. AB - We found a novel octapeptide (H-YRNWFGRW-NH2) mimicking sialyl Lewis X (sLe(X)) carbohydrate from a chemical peptide library with anti-sLe(X) monoclonal antibody (MAb) 2H5. The peptide libraries were constructed by Fmoc-based solid-phase methodology using the mix-split method. The octapeptide sequence was determined by the iterative deconvolution method using anti-sLe(X) MAb 2H5. To define the important residues for interaction with anti-sLe(X) MAb 2H5, alanine-scanning analogues of H-YRNWFGRW-NH2 were synthesized. Substitution of Tyr1, Trp4, Arg7 and Trp8 to Ala resulted in a marked drop in affinity. This result indicates that aromatic and cationic amino residues have a key role in interacting with anti sLe(X) MAb 2H5. The binding property of the octapeptide was evaluated with anti sLe(X) MAb 2H5 and human E-selectin. The octapeptide showed high inhibitory potency (IC50=17.8 nM) for sLe(X) and competitively inhibited the binding of anti sLe(X) MAb 2H5 in a dose-dependent manner. The octapeptide had high affinity (K(d)=0.168 uM) for E-selectin and this binding was inhibited by sLe(X). These results suggest that octapeptide binds to anti-sLe(X) MAb 2H5 or E-selectin at the sLe(X) binding site and sterically interferes with the recognition of anti sLe(X) MAb 2H5 or E-selectin with sLe(X). This peptide may be a useful lead compound for an anti-inflammatory agent targeting selectin. PMID- 21628889 TI - 3-Deoxysappanchalcone inhibits tumor necrosis factor-alpha-induced matrix metalloproteinase-9 expression in human keratinocytes through activated protein-1 inhibition and nuclear factor-kappa B DNA binding activity. AB - Tumor necrosis factor alpha (TNF-alpha), which is a primary cytokine responsible for inflammatory responses in skin, induces the synthesis of matrix metalloproteinase-9 (MMP-9), which causes skin aging. The protective effects of 3 deoxysappanchalcone against TNF-alpha-induced damage was investigated using human skin keratinocytes. The results showed that 3-deoxysappanchalcone inhibited MMP-9 expression at the protein and mRNA level, by blocking the activation of activator protein-1 (AP-1) and nuclear factor kappa B (NF-kappaB). Taken together, the inhibitory activity of 3-deoxysappanchalcone on MMP-9 expression and production in TNF-alpha-treated cells was found to be mediated by the suppression of AP-1 and NF-kappaB activation. PMID- 21628890 TI - Fluorescence investigation of the retinal delivery of hydrophilic compounds via liposomal eyedrops. AB - We examined the feasibility of using submicron-sized liposomes (ssLips) for retinal delivery of hydrophilic compounds, which would also have a wide range of applications. To evaluate the uptake into conjunctival cell line and the intraocular behavior of hydrophilic compound-containing ssLips after eyedrop application, fluorometric investigation was carried out by using a hydrophilic fluorescence probe, 5(6)-carboxyfluorescein (CF). A relatively high amount of CF (>50%) could be incorporated into an internal phase of ssLips by a calcium acetate gradient method. CF being entrapped within the liposomes markedly enhanced both the uptake of CF into conjunctival cells and CF-oriented emission in the retina in mice after eyedrop application, while the free CF did not clear delivery efficiency in both in vitro and in vivo study. In addition, the cellular uptake and luminescence intensity in the retina were higher when a ssLip formulation composed of L-alpha-distearoyl phosphatidylcholine was applied than when a ssLip formulation composed of egg phosphatidylcholine was applied. Consequently, ssLips of appropriate composition were considered to have good potential to carry hydrophilic compounds into the retina. PMID- 21628891 TI - Anti-complementary ginsenosides isolated from processed ginseng. AB - As part of an ongoing search for immunomodulatory components aimed at the anti complementary effect, ginsenosides isolated from processed ginseng were found to have inhibitory activity on complement activation through classical pathways. Activity-guided fractionation was used to isolate four ginsenosides, namely ginsenoside Rg6, F4, Rk3, and Rh4. Ginsenoside Rk3 and Rh4 had a 3 fold higher inhibition activity than rosmarinic acid which was used as a positive control while ginsenoside Rg6 and F4 showed only mild effects similar to that of the positive control. The results suggest that the activity of the corresponding ginsenosides may be increased by the glycosyl moiety at the C6 position rather than the double bond conformation at C20, and ginsenoside Rk3 and Rh4 could have a role in treating inflammatory diseases. PMID- 21628892 TI - Therapeutic effects of hybrid liposomes composed of phosphatidylcholine and docosahexaenoic acid on the hepatic metastasis of colon carcinoma along with apoptosis in vivo. AB - Therapeutic effects of hybrid liposomes (L-alpha-dimyristoylphosphatidylcholine (DMPC)/docosahexaenoic acid (DHA)) composed of 50 mol% DMPC and 50 mol% DHA on the metastasis of human colon carcinoma (HCT116) cells were examined in vivo. DMPC/DHA having a hydrodynamic diameter less than 100 nm were preserved for one month. Remarkably high therapeutic effects were obtained in the hepatic metastasis mouse models of HCT116 cells after the intravenous injection of DMPC/DHA. The histological analysis indicated the induction of apoptosis was observed in the liver section of the hepatic metastasis mouse models treated with DMPC/DHA in vivo. Furthermore, prolonged survival was obtained in the hepatic metastasis mouse models after the treatment with DMPC/DHA. Therapeutic effects of DMPC/DHA without any drugs on the hepatic metastasis were revealed on the basis of histological and biochemical analyses for the first time in vivo. PMID- 21628893 TI - 23-Hydroxytormentic acid and niga-ichgoside f1 isolated from Rubus coreanus attenuate cisplatin-induced cytotoxicity by reducing oxidative stress in renal epithelial LLC-PK1 cells. AB - The unripe fruits of Rubus coreanus (Rosaceae) are used in traditional Chinese medicine to relieve kidney dysfunction. In the present study, we evaluated the protective effects of the triterpenoid glycoside niga-ichigoside F1 (NIF1) and of its aglycone 23-hydroxytormentic acid (23-HTA) isolated from the unripe fruits of Rubus coreanus (Rosaceae) against cisplatin-induced cytotoxicity in renal epithelial LLC-PK1 cells. Pretreating LLC-PK1 cells with 23-HTA or NIF1 was found to prevent cisplatin-induced cytotoxicity and apoptosis. In addition, 23-HTA or NIF1 pretreatment significantly improved the changes associated with cisplatin toxicity by increasing levels of glutathione (GSH) and decreasing levels of malondialdehyde (MDA) and reactive oxygen species (ROS). The activity of antioxidant enzymes including catalase (CAT) and superoxide dismutase (SOD) was significantly lower in cisplatin-treated LL-PK1 cells, and 23-HTA or NIF1 treatment notably increased the these enzyme activity and protein and mRNA levels of CAT and manganese SOD (MnSOD). Moreover, cisplatin caused a significant decrease in nuclear levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and pretreatment with 23-HTA or NIF1 significantly suppressed the cisplatin induced translocation of Nrf2 in LLC-PK1 cells. Taken together, these results suggest that 23-HTA ameliorates cisplatin-induced toxicity via modulation of antioxidant enzymes through activation of Nrf2 in LLC-PK1 cells. PMID- 21628894 TI - Inverse relationship between adipocyte differentiation and ceramide level in 3T3 L1 cells. AB - Adipocyte differentiation has been a target in anti-obesity strategies and is known to be closely related to lipid metabolism. Ceramide, a major sphingolipid metabolite, has been implicated in differentiation. In this study, we investigated whether ceramide biosynthesis is related to adipogenesis in 3T3-L1 cells. Preadipocytes can be differentiated synchronously by a mixture of adipogenic inducers including 3-isobutyl-1-methylxanthine, dexamethasone and insulin. The number of lipid droplets and the triglyceride content, which are differentiation biomarkers, gradually increased during adipogenesis. Interestingly, ceramide and sphingosine contents in the differentiated cells were decreased compared to those in preadipocytes. When the preadipocytes were treated with an 3-isobutyl-1-methylxanthine- or dexamethasone- or insulin-deficient mixture of inducers, the cellular ceramide levels were significantly increased compared with those in cells treated with the complete set of inducers. When preadipocytes were treated with 0, 0.1 or 1 ug/ml insulin along with 3-isobutyl-1 methylxanthine and dexamethasone, the ceramide levels were decreased and the triglyceride content was increased in a concentration-dependent manner. When the cells were treated with epigallocatechin gallate, an adipocyte differentiation inhibitor, during adipogenesis, the ceramide levels of adipocytes were increased and the fat content was decreased. In conclusion, our findings demonstrate that cellular ceramide levels are inversely correlated with adipocyte differentiation. PMID- 21628895 TI - Inhibitory effect of cyclic trihydroxamate siderophore, desferrioxamine E, on the biofilm formation of Mycobacterium species. AB - Formation of biofilm in pathogenic bacteria defends them from antibiotics and the immune system of a host's life. Hence, investigation of the molecular mechanisms of biofilm formation and search for new substances counteracting this formation are becoming an attractive research area. In the course of our search for new inhibitors of biofilm formation in Mycobacterium species, we rediscovered a cyclic trihydroxamate siderophore, desferrioxamine E, from the culture of the marine-derived Actinomycete MS67. Desferrioxamine E inhibited biofilm formation of Mycobacterium smegmatis and M. bovis BACILLE de CALMETTE et GUERIN (BCG) with minimum inhibitory concentration (MIC) value of 10 uM, while no anti-microbial activity was observed up to 160 uM. Desferrioxamine E was also able to restore the anti-microbial activity of isoniazid against M. smegmatis by inhibiting biofilm formation. Mechanistic analysis of desferrioxamine E suggested that such inhibition might come from the depletion of iron in the medium, which is essential for biofilm formation in Mycobacterium species. PMID- 21628896 TI - The functional role of the tachykinin consensus region of urechistachykinin peptide family for its antimicrobial activity. AB - In our previous study, we reported that urechistachykinin I (U I) and II (U II) exerted antimicrobial effects. To find out how the tachykinin consensus sequence of the urechistachykinin peptide family affects its antimicrobial activity, analogues substituting the amino acid residues phenylalanine (Phe-6; Anal 1), glycine (Gly-8; Anal 2), and arginine (Arg-10; Anal 3) of U II to alanine (Ala) were designed. Subsequently, the antimicrobial activity was shown on the order of Anal 3>U II=Anal 2>Anal 1, and this activity pattern was correlated with membrane studies such as propidium iodide (PI) influx and fluorescein isothiocyanate dextran (FD) leakage assay. These results suggest that the antimicrobial activity is related to the hydrophobicity values of the peptides. In regards to the activity of U II, it is determined that the hydrophobic Phe-6 plays a more critical role than Gly-8 or Arg-10. PMID- 21628897 TI - Preventive effect of chlorogenic acid on lens opacity and cytotoxicity in human lens epithelial cells. AB - The present study reveals the pharmacological effect of chlorogenic acid, a major phenolic compound in plants, food, and coffee, on diabetic cataracts. Chlorogenic acid examined the inhibitory effects upon rat lens aldose reductase (AR) activity and xylose-induced rat lens opacity. The effect of chlorogenic acid on high glucose-induced cytotoxicity in lens epithelial cells was also examined. Chlorogenic acid showed potential inhibitory activity against rat lens AR, with an IC50 value of 0.95 uM. The xylose-induced opacity of lenses was significantly improved after treatment with chlorogenic acid in a dose-dependent manner. Chlorogenic acid prevented high glucose-induced cytotoxicity in human lens epithelial (HLE-B3) cells in a dose-dependent manner. These results suggest that chlorogenic acid may provide a potential therapeutic approach for prevention of diabetic complications, such as cataracts. PMID- 21628898 TI - Cross-link formation between mutant galectins of Caenorhabditis elegans with a substituted cysteine residue and asialofetuin via a photoactivatable bifunctional reagent. AB - LEC-1 is the first tandem repeat-type galectin isolated from an animal system; this galectin has two carbohydrate recognition domains in a single polypeptide chain. Because its two lectin domains have different sugar-binding profiles, these domains are thought to interact with different carbohydrate ligands. In our previous study, we showed that a mutant of LEC-1 in which a cysteine residue was introduced at a unique position in the N-terminal lectin domain (Nh) can be cross linked with a model glycoprotein ligand, bovine asialofetuin, by using a bifunctional photoactivatable cross-linking reagent, benzophenone-4-maleimide. In the present work, we applied the same procedure to the C-terminal lectin domain (Ch) of LEC-1. Cross-linked products were formed in the cases of two mutants in which a cysteine residue was introduced at Lys177 and Ser268, respectively. This method is very useful for capturing and assigning endogenous ligand glycoconjugates with relatively low affinities to each carbohydrate recognition domain of the whole tandem repeat-type galectin molecule. PMID- 21628899 TI - Relapse of experimental autoimmune encephalomyelitis after discontinuation of FTY720 (Fingolimod) treatment, but not after combination of FTY720 and pathogenic autoantigen. AB - FTY720 (Fingolimod) is known to have a significant therapeutic effect on experimental autoimmune encephalomyelitis (EAE). Here, we used an EAE mouse model, which had been established by immunizing C57BL/6J mice with a partial peptide of myelin oligodendrocyte glycoprotein (MOG35-55), to examine the relapse of EAE upon discontinuation of treatment with FTY720 alone or in combination with MOG35-55. Relapse was confirmed to occur in all animals (n=6) within one week after discontinuation of FTY720, with increase in the number of lymphocytes infiltrating the spinal cord and demyelination. However, in the case of combination therapy with FTY720 and MOG35-55, relapse following discontinuation of treatment was completely suppressed. The autoantigenic peptide might serve to suppress the clonal selection of relapse-associated autoantigen-specific T cells. PMID- 21628900 TI - Formation mechanism of furfuryl sulfides from o-furfuryl dithiocarbonates: density functional theory study for aromatic [3,3]-sigmatropic rearrangement. AB - Density functional theory (DFT) calculations at the B3LYP/6-31G(d) and B3LYP/6 31G+(d) levels demonstrated that O-furfuryl S-alkyl dithiocarbonate (1) undergoes aromatic [3,3]-sigmatropic rearrangement to the energetically unfavorable S-(2 methylene-2,3-dihydrofuran-3-yl) S-alkyl dithiocarbonate (2'), which then rearranges to furfuryl alkyl sulfide (3) with COS extrusion to regain the aromaticity lost in the first step. PMID- 21628901 TI - Influence of particle design on oral absorption of poorly water-soluble drug in a silica particle-supercritical fluid system. AB - The physicochemical characteristics and oral absorption of a poorly water-soluble drug, K-832, adsorbed onto porous silica (Sylysia 350), were compared with those of K-832 adsorbed onto non-porous silica (Aerosil 200). K-832 and silica were treated with supercritical CO(2) (scCO(2)) to produce K-832-Sylysia 350 and K-832 Aerosil 200 formulations. Scanning electron microscopy, polarizing microscopy, powder X-ray diffraction, and differential scanning calorimetry results suggested that K-832 mainly existed in an amorphous state in both formulations. The specific surface area of both formulations was much larger than that of pure K 832 crystals. The dissolution rate of K-832 from both formulations was considerably greater than that from corresponding physical mixtures due to rapid wetting of the hydrophilic carrier surfaces and amorphous state, the dissolution from the K-832-Sylysia 350 formulation being the fastest. In vivo absorption tests on the two formulations indicated no significant differences in their peak concentration (C(max)) and the area under their plasma concentration-time curve (AUC), while the concentrations of K-832 in the K-832-Sylysia 350 formulation were significantly higher than those in the K-832-Aerosil 200 formulation 1 h and 1.5 h after administration of these formulations (p<0.05). This could be attributed to the different dispersion states of K-832 in the formulations due to their different three-dimensional structures (porous and non-porous). In physical stability tests, the amorphous drugs in both formulations were stable at room temperature for at least 14 months. Thus, the absorption of poorly water-soluble drugs could be greatly improved by adsorption onto porous silica using scCO(2). PMID- 21628902 TI - Microbial metabolism. Part 12. Isolation, characterization and bioactivity evaluation of eighteen microbial metabolites of 4'-hydroxyflavanone. AB - Fermentation of 4'-hydroxyflavanone (1) with fungal cultures, Beauveria bassiana (ATCC 13144 and ATCC 7159) yielded 6,3',4'-trihydroxyflavanone (2), 3',4' dihydroxyflavanone 6-O-beta-D-4-methoxyglucopyranoside (3), 4'-hydroxyflavanone 3'-sulfate (4), 6,4'-dihydroxyflavanone 3'-sulfate (5) and 4'-hydroxyflavanone 6 O-beta-D-4-methoxyglucopyranoside (7). B. bassiana (ATCC 13144) and B. bassiana (ATCC 7159) in addition, gave one more metabolite each, namely, flavanone 4'-O beta-D-4-methoxyglucopyranoside (6) and 6,4'-dihydroxyflavanone (8) respectively. Cunninghamella echinulata (ATCC 9244) transformed 1 to 6,4'-dihydroxyflavanone (8), flavanone-4'-O-beta-D-glucopyranoside (9), 3'-hydroxyflavanone 4'-sulfate (10), 3',4'-dihydroxyflavanone (11) and 4'-hydroxyflavanone-3'-O-beta-D glucopyranoside (12). Mucor ramannianus (ATCC 9628) metabolized 1 to 2,4-trans-4' hydroxyflavan-4-ol (13), 2,4-cis-4'-hydroxyflavan-4-ol (14), 2,4-trans-3',4' dihydroxyflavan-4-ol (15), 2,4-cis-3',4'-dihydroxyflavan-4-ol (16), 2,4-trans-3' hydroxy-4'-methoxyflavan-4-ol (17), flavanone 4'-O-alpha-D-6-deoxyallopyranoside (18) and 2,4-cis-4-hydroxyflavanone 4'-O-alpha-D-6-deoxyallopyranoside (19). Metabolites 13 and 14 were also produced by Ramichloridium anceps (ATCC 15672). The former was also produced by C. echinulata. Structures of the metabolic products were elucidated by means of spectroscopic data. None of the metabolites tested showed antibacterial, antifungal and antiprotozoal activities against selected organisms. PMID- 21628903 TI - Sesquiterpenoids from the Formosan soft coral Lemnalia flava. AB - Four new nardosinane-type sesquiterpenoids, flavalins E-H (1-4) and two new nornardosinane-type norsesquiterpenoids, flavalins I (5) and J (6), along with five known compounds (7-11) have been isolated from a Formosan soft coral Lemnalia flava. The structures of these compounds were elucidated on the basis of their spectroscopic data. Moreover, the absolute configuration of 10 was further determined by Mosher's method. PMID- 21628904 TI - Physico-chemical characterization of gamma-amino n-butyric acid nanoparticles. AB - This work deals with the preparation and characterization of high-purity nanoparticles of gamma-amino n-butyric acid (GABA) in order to enhance the efficiency of this drug. A sublimation procedure at low pressure was applied to GABA after improving the experimental parameters of this physical transformation. The elaboration of the molecule is solvent-free. The process does not change the chemical formula of the compound but modifies its physico-chemical characteristics. In this work, we present the experimental parameters for preparing monoclinic GABA nanoparticles. Their identification and physico chemical properties were determined with a large number of investigations: Powder X-ray diffraction (PXRD), density and purity measurements, differential scanning calorimetry (DSC), thermo-gravimetric analysis (TGA), calorimetric measurements (DeltaH(dissolution) and C(p)), thermally stimulated current (TSC), and electrochemical impedance. PMID- 21628905 TI - Dialkyl bisphosphonate platinum(II) complex as a potential drug for metastatic bone tumor. AB - Bisphosphonates have high affinity for hydroxyapatite (HA), which is abundantly present in bone. Also, platinum complexes are known that have a wide spectrum of antitumor activities. The conjugate of bisphosphonate and a platinum complex might have HA affinity and antitumor activity, and become a drug for metastatic bone tumor. In this study, the authors synthesized platinum complexes that had dialkyl bisphosphonic acid as a ligand, and evaluated the possibility of the synthesized complexes as a drug for metastatic bone tumor. The synthesized dialkyl bisphosphonate platinum(II) complex was characterized, and its stability in an aqueous solution was also confirmed. The synthesized platinum complex showed higher HA affinity than other platinum complexes such as cisplatin and carboplatin in an experiment of adsorption to HA. In vitro, the platinum complex showed tumor growth inhibitory effect stronger than or equal to cisplatin, which is the most commonly used antitumor agent. Moreover, the platinum complex showed a bone absorption inhibitory effect on the osteoclast. These results suggest potential of dialkyl bisphosphonate platinum(II) complexes as a drug for metastatic bone tumor. PMID- 21628906 TI - Regio- and enantioselective allylic substitution with less active N- or O nucleophiles catalyzed by iridium-complex of bis(oxazolinyl)pyridine. AB - The utility of hydroxylamines as nitrogen nucleophiles was investigated in the iridium-catalyzed regio- and enantioselective allylic substitution. Allylic substitution with hydroxylamines proceeded with good enantioselectivities by using the iridium-complex of bis(oxazolinyl)pyridine ligand. The good regio- and enantioselectivities were also achieved in the reaction with alkylamines, p anisidine, and 4-methoxyphenol. PMID- 21628907 TI - Spectrophotometric determination of cobalt(II) and cyanocobalamin with vanillilfluorone and its applications. AB - Spectrophotometric determination of cobalt(II) was accomplished with vanillilfluorone (VF) in the presence of dimethylbenzyltetradecylammonium chloride (Zephiramine, Zep). In the determination of cobalt(II), Beer's law was obeyed in the range of 24-470 ng/ml, with an effective molar absorption coefficient (at 575 nm) and relative standard deviation of 1.35*10(5) l mol(-1) cm(-1) and 0.66% (n=5), respectively. The composition ratio of the colored complex was determined by the mole ratio and continuous variation methods, and it was found to be Co(II) : VF : Zep=1 : 2 : 4. Analysis of cyanocobalamin by the same procedure showed that cyanocobalamin could be determined in the concentration range of 0.5-0.11 ug/ml using the proposed method. PMID- 21628908 TI - Three new flavonoid glycosides, byzantionoside B 6'-O-sulfate and xyloglucoside of (Z)-hex-3-en-1-ol from Ruellia patula. AB - Three new flavonoid glycosides, demethoxycentaureidin 7-O-beta-D galacturonopyranoside, pectolinarigenin 7-O-alpha-L-rhamnopyranosyl-(1'''->4") beta-D-glucopyranoside and 7-O-alpha-L-rhamnopyranosyl-(1'''->4")-beta-D glucuronopyranoside, a new megastigmane glucoside, byzantionoside B 6'-O-sulfate, and a new (Z)-hex-3-en-1-ol O-beta-D-xylopyranosyl-(1"->2')-beta-D glucopyranoside, were isolated from leaves of Ruellia patula JACQ., together with 12 known compounds, beta-sitosterol glucoside, vanilloside, bioside (decaffeoyl verbascoside), acteoside (verbascoside), syringin, benzyl alcohol O-beta-D xylopyranosyl-(1"->2')-beta-D-glucopyranoside, cistanoside E, roseoside, phenethyl alcohol O-beta-D-xylopyranosyl-(1"->2')-beta-D-glucopyranoside, (+) lyoniresinol 3alpha-O-beta-D-glucopyranoside, isoacteoside and 3,4,5 trimethoxyphenol O-alpha-L-rhamnopyranosyl-(1"->6')-beta-D-glucopyranoside. Their structures were elucidated by means of spectroscopic analyses. PMID- 21628909 TI - A new series of 2-alkoxy(aralkoxy)-[1,2,4]triazolo[1,5-a]quinazolin-5-ones as adenosine receptor antagonists. AB - This research was carried out to study the pharmacological activity of a newly synthesized series of 2-alkoxy-[1,2,4]triazolo[1,5-a]quinazolin-5-ones as adenosine receptor antagonists. These compounds have been tested in radioligand binding assays on cloned Chinese hamster ovary (CHO) cells transfected with A(1), A(2A), A(2B) and A(3) receptors. In particular, among the triazoloquinazolines (1 11), the dialkoxy derivative (7b) was found to have the highest affinity at A(1) subtype receptor, and its radioligand binding activity together with 1,3-dipropyl 8-cyclopentylxanthine (DPCPX) was studied. Finally, the structure-activity relationship (SAR) studies on the titled compounds provide some new insights about steric hindrance and lipophilic requirements for anchoring to the adenosine receptors recognition site. PMID- 21628910 TI - Interaction characteristics with bovine serum albumin and retarded nitric oxide release of ZCVI4-2, a new nitric oxide-releasing derivative of oleanolic acid. AB - ZCVI4-2 is a newly developed furoxan-based nitric oxide-releasing derivative of oleanolic acid. It exhibited strong cytotoxicity against human hepatocellular carcinoma (HCC) in vitro and significantly inhibited the growth of HCC tumors in vivo. However, its low aqueous solubility and toxicity due to the fast release of nitric oxide (NO) in blood challenged its formulation. In the present investigation, the interaction characteristics of ZCVI4-2 with bovine serum albumin (BSA) were studied by fluorescence spectrometry, synchronous fluorescence spectra and Fourier transform-infrared (FT-IR). It was found that ZCVI4-2 concentration, temperature and pH had significant effect on the interactions. ZCVI4-2 was able to bind BSA with high affinity, low temperature and neutral pH favor the binding. The interaction exhibited to be a spontaneous and exothermic process. ZCVI4-2 was buried in the hydrophobic pocket in subdomain IIB of BSA and the exact binding site was around 3.83 nm in average from Trp212. The NO releasing characteristics of nanocomplexes were compared with ZCVI4-2 solution by Griess Reagent Method. It was found that the release of NO from ZCVI4-2/BSA nanocomplexes was retarded significantly, thus making ZCVI4-2 into a BSA-bound nanocomplexes had the great potential to lower the toxicity due to the absence of organic solvents and surfactants and meanwhile the sustained release of NO. PMID- 21628911 TI - A new polyoxygenated triterpene and two new aeginetic acid quinovosides from the roots of Rehmannia glutinosa. AB - A new minor polyoxygenated triterpene named glutinolic acid (1) and two new aeginetic acid quinovosides (2, 3) were isolated from the roots of Rehmannia glutinosa LIBOSCH. (Scrophulariaceae) cultivated in Gunwi-gun, Korea. The structures of these compounds were established as 3alpha,19alpha,20beta,24,30 pentahydroxyurs-12-en-28-oic acid (1, glutinolic acid), aeginetic acid 5-O-beta-D quinovoside (2) and aeginetoyl ajugol 5"-O-beta-D-quinovoside (3) on the basis of chemical and spectroscopic evidence. PMID- 21628912 TI - n-Octyl alpha-L-rhamnopyranosyl-(1->2)-beta-D-glucopyranoside derivatives from the glandular trichome exudate of Geranium carolinianum. AB - Chemical investigation of the glandular trichome exudate from Geranium carolinianum L. (Geraniaceae) led to the characterization of unique disaccharide derivatives, n-octyl 4-O-isobutyryl-alpha-L-rhamnopyranosyl-(1->2)-6-O-isobutyryl beta-D-glucopyranoside (1), n-octyl 4-O-isobutyryl-alpha-L-rhamnopyranosyl-(1->2) 6-O-(2-methylbutyryl)-beta-D-glucopyranoside (2) and n-octyl 4-O-(2 methylbutyryl)-alpha-L-rhamnopyranosyl-(1->2)-6-O-isobutyryl-beta-D glucopyranoside (3), named caroliniasides A-C, respectively. These structures were determined by spectral means. n-Alkyl glycoside derivatives have been isolated from the glandular trichome exudates for the first time. This rare type of secondary metabolites could be applicable to chemotaxonomic perspective because they are found in glandular trichome exudates of plants belonging to the genus Geranium, according to our studies. PMID- 21628913 TI - Synthesis of biologically active (-)-dehydroiso-beta-lapachone and the determination of its absolute configuration. AB - Synthesis of dehydoriso-beta-lapachone (1) in both racemic and enantioenriched forms is achieved starting from reduced naphthoquinone equivalents. As for the synthesis of enantioenriched dehydroiso-beta-lapachone, introduction of the asymmetric center was carried out by catalytic asymmetric epoxidation of the unfunctionalized trisubstituted olefin using Shi epoxidation diketal catalyst. The construction of isopropenylfurano-1,2-(beta)-naphthoquinone was carried out by acidic ring-opening reaction of the epoxynaphthalene and the following diammonium cerium(IV) nitrate (CAN) oxidation. The absolute configuration of naturally occurring (-)-dehydroiso-beta-lapachone was finally determined as (R) by comparing the measured optical rotation value of the synthetic (R)-dehydroiso beta-lapachone. PMID- 21628914 TI - Chromanols from Sargassum siliquastrum and their antioxidant activity in HT 1080 cells. AB - Six meroterpenoids (compounds 1-6) of chromene class, including three known compounds (1-3), were isolated from Sargassum siliquastrum. The structure of these compounds was established by extensive 2D-NMR experiments such as (1)H gradient double quantum filtered correlation spectroscopy (gDQCOSY), total correlation spectroscopy (TOCSY), nuclear Overhauser effect spectroscopy (NOESY), gradient heteronuclear multiple quantum coherence (gHMQC), and gradient heteronuclear multiple bond correlation (gHMBC), and by comparison with published spectral data. The antioxidant activity of these compounds was evaluated by various antioxidant tests, such as scavenging effects on generation of intracellular reactive oxygen species (ROS), increments of intracellular glutathione (GSH) level, and inhibitory effects on lipid peroxidation in human fibrosarcoma HT 1080 cells. Compounds (1-6) significantly decreased generation of intracellular ROS and inhibited lipid peroxidation while they increased levels of intracellular GSH at a concentration of 5 ug/ml. PMID- 21628915 TI - Drimane sesquiterpenoids from the mangrove-derived fungus Aspergillus ustus. AB - Five new drimane sesquiterpenes (1-5) together with 14 known analogues (6-19) were isolated from laboratory cultures of a mangrove-derived fungus Aspergillus ustus. Their structures were established by spectroscopic methods and antitumor activities were evaluated by sulforhodamine B (SRB) and 3-(4,5-dimethylthiazol-2 yl)-2,5-diphenyltetrazolium bromide (MTT) methods. PMID- 21628916 TI - Composite constituent: lactucenyl acetate, a novel migrated lupane triterpenoid from Lactuca indica revision of structure of tarolupenyl acetate. AB - Lactucenyl acetate (1), a new member of migrated lupane triterpenoids was isolated from Lactuca indica and its structure was elucidated on the basis of spectral analyses. The structure of tarolupenyl acetate was revised as lup-19(21) en-3beta-yl acetate (2). PMID- 21628917 TI - Antioxidant hispidin derivatives from medicinal mushroom Inonotus hispidus. AB - Two natural antioxidants, named inonotusin A (1) and B (2), were isolated from the methanolic extract of the fruit bodies of Inonotus hispidus, together with (E)-4-(3,4-dihydroxyphenyl)but-3-en-2-one (3), hispidin (4) and 3,4 dihydroxybenzaldehyde (5). Their structures were identified by means of extensive NMR and MS data analysis. Compounds 1, 2 and 4 exhibited significant scavenging activity against the 2,20-azinobis(3-ethylbenzhiazoline-6-sulfonate) (ABTS) radical cation. Compound 1 also showed moderate cytotoxicity against a human breast carcinoma cell line (MCF-7) with IC(50) values of 19.6 uM. PMID- 21628918 TI - Three new megastigmane glucopyranosides from the Cardamine komarovii. AB - Three new megastigmane glucopyranosides, komaroveside A [(3S,4R,5Z,7E)-3,4 dihydroxy-5,7-megastigmadien-9-one-3-O-beta-D-glucopyranoside] (1), komaroveside B [(3S,4S,5S,6R,7E)-5,6-epoxy-3,4-dihydroxy-7-megastigmen-9-one-3-O-beta-D glucopyranoside] (2) and komaroveside C [(3S,4S,5S,6R,7E,9S)-5,6-epoxy-3,4,9 trihydroxy-7-megastigmen-3-O-beta-D-glucopyranoside] (3) were isolated, together with eight known compounds, from Cardamine komarovii. The identification of these compounds and the elucidation of their structures were based on 1D- and 2D-NMR spectral data analysis. The isolated compounds were tested for their cytotoxicity against four human tumor cell lines (A549, SK-OV-3, SK-MEL-2, HCT15) in vitro using the sulforhodamine B bioassay. PMID- 21628919 TI - New triterpenes from Barringtonia asiatica. AB - The leaves of Barringtonia asiatica afforded two new triterpenes, germanicol caffeoyl ester (1) and camelliagenone (2). Their structures were elucidated by extensive 1D- and 2D-NMR spectroscopy. It also afforded germanicol trans coumaroyl ester (3), germanicol cis-coumaroyl ester (4), germanicol (5), camelliagenin A (6), spinasterol, sitosterol, squalene, lutein and trilinolein. Compounds 3, spinasterol and trilinolein were isolated from the fruits, while the seeds yielded spinasterol, squalene, linoleic acid and trilinolein. Compounds 1-5 exhibited antifungal activity against Candida albicans, 1-3 and 5 showed antibacterial activity against Staphylococcus aureus, while 5 is active against Pseudomonas aeruginosa. PMID- 21628920 TI - Synthesis of N-(trifluoromethyl-2-pyridinyl)arenesulfonamides as an inhibitor of secretory phospholipase A2. AB - A series of N-(trifluoromethyl-2-pyridinyl)alkane- and arenesulfonamides 2-5 have been synthesized by the substitution reaction of 2 chloro(trifluoromethyl)pyridines 6 with alkane- and arenesulfonamides 7. Their inhibitory activities against secretory phospholipase A2 of porcine pancreas were examined and the analog N-[4,5-bis(trifluoromethyl)-2-pyridinyl]-4 trifluoromethylbenzenesulfonamide 4i was shown to have the highest inhibitory activity, with an IC(50) value of 0.58 mM. PMID- 21628921 TI - Synthesis of renieramycins: construction of the core ring system of cribrostatin 4 through modified Pictet-Spengler cyclization of 3,6-bisarylpiperazine-2,5-dione with diethoxyethyl benzoate. AB - A nine-step synthesis of pentacyclic key intermediate 11 of cribrostatin 4 (2) along with renieramycin I (1i) from 3,6-bisarylpiperazine-2,5-dione derivative 3 is described. The key step of this synthesis is the stereoselective cyclization of lactam nitrogen with diethoxyethyl benzoate, followed by the stereoselective hydrogenation to generate ABC ring system 6. PMID- 21628922 TI - Efficient topical delivery of chlorogenic acid by an oil-in-water microemulsion to protect skin against UV-induced damage. AB - We examined the intradermal delivery of a hydrophilic polyphenol chlorogenic acid by in vitro study using excised guinea pig dorsal skin and Yucatan micropig skin. Skin accumulation as well as the solubility of chlorogenic acid in aqueous vehicles was much greater than for other polyphenols such as quercetin and genistein. However, since enhancement of skin delivery seemed to be necessary to exhibit its protective effects against oxidative damage of skin, we examined the effects of microemulsions as vehicles. Using microemulsions consisting of 150 mM NaCl solution, isopropyl myristate, polyoxyethylene sorbitan monooleate (Tween 80) and ethanol, skin accumulation as well as solubility of chlorogenic acid further increased. Enhancement effect of an oil-in-water (o/w-type) microemulsion was greater than that of a water-in-oil (w/o-type) microemulsion possibly due to the greater increase in solubility. This finding was quite different from previous findings on relatively hydrophobic polyphenols such as quercetin and genistein. Pretreatment of guinea pig dorsal skin with chlorogenic acid containing microemulsion gel prevented erythema formation induced by UV irradiation. These findings indicate the potential use of hydrophilic chlorogenic acid with o/w-type microemulsion as a vehicle to protect skin against UV-induced oxidative damage. PMID- 21628923 TI - Practical preparation of ethyl 2-methylthiophene-3-carboxylate. AB - A safe and efficient process for the preparation of ethyl 2-methylthiophene-3 carboxylate (5) was devised. This process provides several advantages over the precedents, involving operational simplicity, avoidance of the use of strong bases such as n-butyllithium and application of noncryogenic conditions, and enabled to prepare 5 in 52% overall yield from commercially available 2 methylthiophene on a multikilogram scale. PMID- 21628924 TI - A simple, iron-catalyzed, pyridine-assisted hydrogen peroxide epoxidation system. AB - A simple and inexpensive system comprised of H(2)O(2)-pyridine-FeCl(3).6H(2)O for the catalysis of olefin epoxidation was established. Intriguingly, the reactivity of this system greatly depends on the amounts of pyridine. Various substrates, including aromatic and aliphatic olefins, were epoxidized by this simple system in moderate to excellent yields. PMID- 21628925 TI - Diagnosis of gallbladder tumors. AB - Gallbladder cancer is among the organs examined in mass screening for cancer using ultrasonography; the reported prevalence of gallbladder cancer in such screening of a general population was 0.011%, while the prevalence of gallbladder polyps was reported as 4.3 to 6.9%. Endoscopic ultrasonography is useful for the differential diagnosis of gallbladder tumors detected by mass screening, as well as for estimating the depth of tumor invasion and detecting abnormal connections between pancreatobiliary ducts. While a systematic approach leading to diagnosis by endoscopic ultrasonography is useful, recent advances of contrast-enhanced ultrasonography are expected to establish it as a new modality for early detection. At our hospital, 7 of 26 patients with abnormal connections between pancreatobiliary ducts developed gallbladder carcinoma (23.1%), and 7 of 48 patients with gallbladder carcinoma had abnormal connections between pancreatobiliary ducts (12.5%). Serial observation in patients with gallstones and prophylactic surgery in patients with abnormal connections between pancreatobiliary ducts are necessary. PMID- 21628926 TI - Predictive value of the fibrosis scores in patients with chronic hepatitis C associated with liver fibrosis and metabolic syndrome. AB - OBJECTIVE: We evaluated patients with chronic hepatitis C (CHC) and compared the clinical and pathological features of steatosis and metabolic syndrome to identify the risk factors for CHC with severe fibrosis. METHODS: One hundred seventy-one patients with biopsy-confirmed CHC were included in the study: 90 males and 81 females, age 56.2 +/- 12.8 years; 46 with obesity (BMI>=25 kg/m(2)); 51 with hypertension; 36 with type 2 diabetes mellitus; and 20 with hypertriglyceridemia. RESULTS: Steatosis was detected in 79 patients (46%); 92 patients (54%) showed no steatosis. Seventy-four patients (43%) showed mild fibrosis and 97 patients (56%) showed severe fibrosis. The variables that were significantly associated with steatosis were obesity [odds ratio 2.160 (1.010 4.727), p=0.046] and type 2 diabetes [odds ratio 3.667 (1.559-8.430), p=0.027]. The variables that were significantly associated with severe fibrosis were older age [odds ratio 2.675 (1.309-5.464), p=0.007], obesity [odds ratio 2.156 (1.006 4.619), p=0.048] and type 2 diabetes [odds ratio 8.739 (2.845-26.846), p=0.0002]. Nagasaki (N) score (the total number of specific risk factors, namely an older age, obesity, and type 2 diabetes) was higher in the severe fibrosis group than in the mild fibrosis group (mild fibrosis: severe fibrosis=1.48 +/- 1.14 vs.2.66 +/- 94, p<0.001). CONCLUSION: Metabolic syndrome factors, including obesity and diabetes, play a critical role in the pathogenesis of fibrosis in CHC. The N score was therefore found to be a significant predictor of severe fibrosis in CHC. PMID- 21628927 TI - Association of amino-terminal pro-brain natriuretic peptide with metabolic syndrome. AB - OBJECTIVE: This study evaluated the relationship between individual components in metabolic syndrome (MetS) and amino-terminal pro-brain natriuretic peptide (NT proBNP). METHODS: A screening program for MetS in 2008 in Taiwan excluded subjects aged <30 years and pregnant women. Fasting glucose, insulin level, high sensitivity C-reactive protein (hsCRP), and NT-proBNP were assessed. A propensity score matching process was used to select subjects with and without MetS comparable in age, gender, body height, and serum creatinine levels. A multiple regression model was used to determine the association between individual components of MetS and NT-proBNP. Finally 270 subjects with MetS and another 270 matched subjects without MetS aged >=30 years were included. RESULTS: The subjects with MetS had higher uric acid and hsCRP, but not NT-proBNP. Multiple regression model showed that log (NT-proBNP) was positively associated with systolic blood pressure (beta=0.002 per mmHg, p=0.013), but negatively associated with body mass index (beta=-0.017 per kg/m(2), p=0.018), triglyceride (beta= 0.00048 per mg/dL, p=0.020) and insulin level (beta=-0.005 per mU/L, p=0.005). Log (NT-proBNP) was neutral to waist circumference, fasting glucose, high-density lipoprotein cholesterol, and diastolic blood pressure. CONCLUSION: MetS was not associated with serum NT-proBNP concentrations due to the contradictory effects of each component. PMID- 21628928 TI - Achievement status toward goal blood pressure levels and healthy lifestyles among Japanese hypertensive patients; cross-sectional survey results from Fukushima Research of Hypertension (FRESH). AB - OBJECTIVE: To determine success rates in achieving goal blood pressures by measuring several factors including lifestyle characteristics associated with therapeutic failures for target treatment goals. METHODS AND PATIENTS: This prospective observational cohort study (September 2008 to September 2010) collected information on blood pressure control status and healthy lifestyle practices listed in Belloc and Breslow's seven health practices through medical records and patients' self-administrated questionnaires among 1,264 of 1,753 Japanese hypertensive patients initially registered. Multivariate analysis was performed to estimate the association between lifestyle practices and "therapeutic failures" at the baseline survey. RESULTS: Median age and proportion of males were 74 years and 41.1%. Therapeutic failures were 68.3% in non-elderly patients (<65 years of age) without diabetes mellitus or chronic kidney disease, 62.6% in patients with these diseases or those who had myocardial infarction, 41.8% in elderly patients (>=65 years of age) without these diseases and only 28.6% in patients with cerebrovascular disease. The total number of healthy lifestyle items was inversely associated with therapeutic failures in multivariate analysis among both sexes. CONCLUSION: The study revealed low achievement rates toward treatment goals, especially non-elderly patients without diabetes mellitus or chronic kidney disease, and patients with these diseases or those who had myocardial infarction. Our data indicated the necessity to improve physicians' awareness of the management of hypertension according to treatment guidelines and the importance of a healthy lifestyle to maintain goal blood pressure levels. PMID- 21628929 TI - Efficacy of blood-patch pleurodesis for secondary spontaneous pneumothorax in interstitial lung disease. AB - OBJECTIVE: We identified the prognostic relevance of pneumothorax in interstitial lung disease (ILD) patients and evaluated the efficacy and safety of autologous blood-patch pleurodesis. METHODS: We retrospectively reviewed 59 occurrences of pneumothorax in 34 ILD patients identified over a 12-year period. RESULTS: Air leakage ceased in 16 of 22 (72.7%) episodes after blood pleurodesis and in 11 of 14 (78.6%) episodes after chemical pleurodesis. Both the cure ratio and recurrence ratio in the cure episodes were comparable with those in the chemical pleurodesis group (p=0.99 and 0.99, respectively). In addition, there were no harmful events associated with blood pleurodesis. The median survival time after the first episode of pneumothorax was less than 9 months in patients with idiopathic interstitial pneumonia (IIP) and only around 3 years in the patients with other types of ILD, which have essentially favorable outcomes. Kaplan-Meier survival estimates were significantly worse in the patients with concomitant pneumomediastinum than in those without (p<0.05). A multivariate Cox regression analysis identified that the number of episodes of pneumothorax, IIP diagnosis and concomitant pneumomediastinum were independent predictors of death. CONCLUSION: Autologous blood-patch pleurodesis is safe and worth considering as a first-line treatment for pneumothorax secondary to ILD. However, despite treatments, the prognosis after the onset of pneumothorax in ILD patients was found to be poor. In addition, concomitant pneumomediastinum may further worsen the prognosis. PMID- 21628930 TI - Clinical study of the time course of clinical symptoms of Pandemic (H1N1) 2009 influenza observed in young adults during an initial epidemic in Kobe, Japan. AB - OBJECTIVE: Although the rates of reported symptoms of Pandemic (H1N1) 2009 influenza virus infection are well studied, the course of progression of these symptoms is not clear. In this study, we carefully reviewed the progress of each patient after hospitalization and clarified the clinical course of the symptoms. METHODS: We retrospectively examined the clinical data of 16 consecutive patients who had been hospitalized during the early stages of an influenza epidemic and observed the clinical progression of their symptoms. RESULTS: Each symptom had a different time of onset and progression pattern. In roughly one-third of our patients, symptoms appeared before the onset of high fever. Acute respiratory symptoms tended to last longer than other symptoms; similarly, sore throat and cough lasted longer than rhinorrhea. The SpO(2) of the patients with influenza showed a declining trend. The point at which minimum SpO(2) levels were noted was approximately 1.5 days after onset of fever. CONCLUSION: In this H1N1 epidemic, patients typically tended to experience general fatigue, sore throat, and cough before the onset of fever, with sore throat and cough lasting longer than the other symptoms. Most patients showed decreased SpO(2) levels at -1.5 days after onset of fever. PMID- 21628931 TI - Clinical evaluation of endoscopic bronchial occlusion with silicone spigots for the management of persistent pulmonary air leaks. AB - OBJECTIVE: The aim of this study was to evaluate the clinical effectiveness of endoscopic bronchial occlusion (EBO) with endobronchial Watanabe spigots (EWSs), a type of silicone bronchial blocker, for managing prolonged pulmonary air leaks. PATIENTS AND METHODS: Between October 2002 and April 2010, 24 patients with surgically incurable pulmonary air leaks underwent EBO with EWSs. The spigot was grasped with forceps and inserted into the affected bronchus by using a flexible bronchoscope through an endotracheal or a tracheostomy tube. RESULTS: In each patient, at least one EWS (mean=2.8) was placed for air leaks due to pneumothorax (n=15), empyema (n=8), or postsurgical complications (n=1). Twelve patients (50%) had complete resolution of the air leaks and seven (29.2%) had a reduction in air leaks, but five (20.8%) showed no improvement. Twenty-three patients required thoracic drainage tubes, which were successfully removed after EBO in 15 patients (65.2%). Of the 24 patients, four experienced severe respiratory failure requiring mechanical ventilation but were successfully treated. Complications were spigot migration, atelectasis, pneumonia, and lung abscess, but none caused significant mortality. CONCLUSION: EBO with EWSs seems to be a reasonable and manageable treatment option for patients with prolonged pulmonary air leaks, including those with severe respiratory failure requiring mechanical ventilation. PMID- 21628932 TI - Autosomal dominant late-onset quadriceps myopathy: three patients of a Taiwanese kindred. AB - OBJECTIVE: Primary quadriceps weakness/atrophy is a rare disorder with variable etiologies; therefore, this disorder has been regarded as a clinical syndrome rather than a distinct entity. However, three affected patients of a Taiwanese family demonstrate a uniform pattern of quadriceps weakness and atrophy, their clinical manifestations and pattern of inheritance may suggest a new disease entity. PATIENTS AND METHODS: Three patients in a Taiwanese kindred with selective quadriceps weakness and atrophy, which began after age 40 years, were examined. To disclose the confines of this disorder, muscle CT scans, electromyography, nerve conduction studies and muscle biopsies were performed; and to unravel and better understand the nature of this disorder, histopathological, ultrastructural, immunocytochemical and genetic studies were carried out. RESULTS: In two patients with long-standing disease, muscle imaging showed marked atrophy and fat replacement of the anterior thigh muscles and electromyography showed a mixture of myopathic and neuropathic changes. Muscle histopathology on the mildly affected tibialis anterior showed myopathic changes with myofibrillar degeneration and secondary neurogenic alterations. Immunocytochemical staining was not diagnostic but excluded the dystrophinopathies and other well-known muscular dystrophies. CONCLUSION: All previously identified diseases resulting in quadriceps weakness and atrophy have been ruled out and the present disorder appears to be a new disease entity of autosomal dominant late onset quadriceps myopathy. PMID- 21628933 TI - Evaluation of an assessment system for the JRS 2005: A-DROP for the management of CAP in adults. AB - OBJECTIVE: The Japanese Respiratory Society (JRS) last revised the guidelines for community-acquired pneumonia (CAP) in adults in 2005. These guidelines proposed new criteria (A-DROP) to assess the severity of pneumonia and to differentiate between typical bacterial pneumonia and atypical pneumonia. The goal of the present study was to evaluate the utility of the A-DROP criteria for these described purposes. METHODS: An observational survey was conducted between July 2006 and March 2007, and patients with CAP were prospectively surveyed using consecutive enrollment methods. PATIENTS: In total, 1,875 patients from 200 medical facilities throughout Japan were analyzed. RESULTS: The JRS 2005 A-DROP system was a good indicator of mortality in the patient population, and these results were significantly correlated with the Pneumonia Severity Index (PSI) of the Infectious Disease Society of America (IDSA). Among the various factors characterized, 'SpO(2) of 90% or less (PaO(2) of 60 Torr or less)' was the strongest predictor of mortality. In terms of the differential diagnosis between typical bacterial and atypical pneumonia, five of six JRS 2005 items were strongly and significantly correlated with a diagnosis of atypical pneumonia. CONCLUSION: The JRS 2005 A-DROP system was accurate and clinically useful for the assessment of the severity of pneumonia and for the differentiation between typical bacterial pneumonia and atypical pneumonia. PMID- 21628934 TI - Focal nodular hyperplasia-like nodule with reduced expression of organic anion transporter 1B3 in alcoholic liver cirrhosis. AB - We report a patient with alcoholic liver cirrhosis who had a 15 mm focal nodular hyperplasia (FNH)-like nodule in the liver. This FNH-like nodule was diagnosed as hepatocellular carcinoma (HCC) mainly based on hypervascularity during the hepatic arterial phase, washout pattern during the equilibrium phase and low signal intensity during the hepatobiliary phase in gadolinium-ethoxybenzyl diethylenetriamine pentaacetic acid (Gd-EOB-DTPA)-enhanced MRI; it was surgically resected. Its histology exhibited hepatocyte hyperplasia, fibrous septa containing unpaired small arteries accompanied by reactive bile ductules, remarkable iron deposits and sinusoidal capillarization, and was compatible with the diagnosis of an FNH-like nodule. When we analyzed the images of the present nodule retrospectively, low signal intensity on in-phase and isosignal intensity on opposed-phase T1-weighted MRI may have reflected iron deposits in the FNH-like nodule. In addition, a low signal intensity on T2-weighted MRI and no detection in diffusion-weighted MRI may help in distinguishing FNH-like nodules from HCC, since these image findings are inconsistent with typical HCC. Immunohistochemical analysis revealed a markedly reduced expression of organic anion transporter (OATP) 1B3 in this nodule, which implied decreased Gd-EOB-DTPA uptake by hepatocytes and accounted for the low signal intensity during the hepatobiliary phase on Gd-EOB-DTPA-enhanced MRI. To the best of our knowledge this is the first report in which an FNH-like nodule was assessed for OATP1B3 expression. PMID- 21628935 TI - Ventricular tachycardia in cardiac sarcoidosis controlled by radiofrequency catheter ablation. AB - We report a case of a 78-year-old woman with cardiac sarcoidosis with a history of syncope and palpitation. Further assessment with echocardiography, gadolinium enhanced cardiovascular magnetic resonance (CMR) and histology led to a diagnosis of cardiac sarcoidosis. As the patient suffered from ventricular tachycardia (VT) despite active corticosteroid therapy, an implantable cardioverter-defibrillator (ICD) was positioned. She was also administered a beta blocker, but an electrical storm appeared every several days requiring ICD therapy. The drug-refractory VT was finally controlled with a catheter ablation session, during which we could detect the VT focus in the right ventricular outflow tract next to the aneurysm by using an electroanatomic mapping system (CARTO). Referring to echocardiographic and CMR images proved very useful in detecting the aneurysm using the CARTO system. PMID- 21628936 TI - Early detection of cardiac sarcoid lesions with (18)F-fluoro-2-deoxyglucose positron emission tomography. AB - In April 2005, a 72-year-old woman with pulmonary sarcoidosis exhibited focal (18)F-fluoro-2-deoxyglucose ((18)F-FDG) uptake in her heart on (18)F-FDG positron emission tomography (PET). Although Japanese guidelines for diagnosing cardiac sarcoidosis were not met at this point, electrocardiography, echocardiography, and magnetic resonance imaging became diagnostic for cardiac sarcoidosis 1 year later. In the present case report, the potential of (18)F-FDG PET in the early recognition of cardiac sarcoidosis in comparison with other imaging modalities is discussed. PMID- 21628937 TI - Inheritance of an autosomal recessive disorder, Gitelman's syndrome, across two generations in one family. AB - Gitelman's syndrome (GS) is an autosomal recessive disorder; it is rarely inherited over several generations. A 16-year-old boy showed hypokalemia and hypocalciuria. Clinically, he was diagnosed as GS because of diuretic responsiveness to furosemide but not thiazide. Genetic testing disclosed he was a compound heterozygote (T180K/V677M) for the SLC12A3 gene. Unexpectedly, the patient's father also showed hypokalemia and hypocalciuria. The genetic analysis showed he had an L849H mutation in addition to T180K. The present pedigree showed an extremely rare case. Diuretic tests are useful diagnostic methods, and genetic testing is necessary for precise evaluation of complicated cases as in this family. PMID- 21628938 TI - Diffuse large B-cell lymphoma complicated with autoimmune thrombocytopenia. AB - A patient was diagnosed with autoimmune thrombocytopenia and concurrent diffuse large B-cell lymphoma (DLBCL). Both autoimmune thrombocytopenia and DLBCL were successfully treated using chemotherapy, while steroid therapy, rituximab monotherapy, and high-dose parenteral glucocorticoid therapy combined with high dose intravenous immunoglobulin failed to improve thrombocytopenia. A greater understanding of the pathogenesis and treatment options can only be definitively clarified by analyses of settings such as the present case, as autoimmune thrombocytopenia is uncommon in patients with malignant lymphoma and the etiology and optimal treatment remain unclear. PMID- 21628939 TI - Fatal encephalitis in a case of hypereosinophilic syndrome: MRI and autopsy findings. AB - A 34-year-old man developed fever and headache, followed by finger tremor and gait disturbance, and was admitted to our hospital about two months after onset. Blood tests showed a white blood cell count of 32,600 /uL with an eosinophil count of 22,300 /uL. There was no evidence of allergic drug reaction or parasitic infection. Cerebrospinal fluid examination demonstrated mononuclear pleocytosis without eosinophils or atypical cells. Brain MRI showed symmetric lesions bilaterally in the medial temporal lobe, frontobasal and insular regions and medulla oblongata. Herpes simplex virus-DNA was negative in the cerebrospinal fluid. The patient died about four months after onset. Histopathologically, there was infiltration of T cells, B cells and macrophages throughout the whole brain, but eosinophils or atypical cells were absent. Immunohistochemistry for herpes simplex virus type 1 and human herpesvirus 6 was negative. This case suggests that fatal encephalitis may develop in association with hypereosinophilic syndrome. PMID- 21628940 TI - Reversible cerebral vasoconstriction syndrome presenting as subarachnoid hemorrhage, reversible posterior leukoencephalopathy, and cerebral infarction. AB - Reversible cerebral vasoconstriction syndrome (RCVS) is characterized by acute severe headache with or without additional neurological symptoms and reversible cerebral vasoconstriction. Unruptured aneurysm has been reported in some cases with RCVS. We report a severe case of a 53-year-old woman with RCVS having an unruptured cerebral aneurysm and presenting as cortical subarachnoid hemorrhage, reversible posterior leukoencephalopathy syndrome, and cerebral infarction. She was successfully treated with corticosteroids and a calcium channel blocker and the aneurysm was clipped. Her various complications are due to the responsible vasoconstriction that started distally and progressed towards proximal arteries. This case demonstrates the spectrum of presentations of RCVS, a clinically complicated condition. PMID- 21628941 TI - Reading epilepsy as the initial symptom of idiopathic hypoparathyroidism. AB - Reading epilepsy is a rare form of reflex epilepsy that occurs exclusively during reading. In this report, we describe a patient in whom seizure was the initial symptom of idiopathic hypoparathyroidism. A 31-year-old man experienced seizures three times in different bookshops while browsing. Chvostek's sign was observed. Serum calcium and parathyroid hormone levels were decreased. Brain computed tomography revealed symmetrical calcification at the dentate nuclei of the cerebellum and the basal ganglia. The interictal electroencephalography was normal. There was no photoparoxysmal response. After amelioration of serum calcium level with oral administration of vitamin D, seizure attacks ceased without antiepileptic drugs. This case is, to our knowledge, the first report of reading epilepsy associated with hypocalcemia due to idiopathic hypoparathyroidism. The psychological stress of reading at bookshops without buying might have triggered seizures under CNS hyperexcitability due to hypocalcemia. PMID- 21628942 TI - IgG4-related tubulointerstitial nephritis and hepatic inflammatory pseudotumor without hypocomplementemia. AB - Immunoglobulin G4 (IgG4)-related tubulointerstitial nephritis (TIN) is often accompanied by autoimmune pancreatitis (AIP) or chronic sclerosing dacryoadenitis and sialoadenitis. However, IgG4-related TIN without AIP or lacrimal and/or salivary gland lesions has not been well recognized. Here, we report a case of IgG4-related TIN associated with hepatic inflammatory pseudotumor without AIP or lacrimal and/or salivary gland lesions. A 58-year-old Japanese man with epigastralgia underwent contrast-enhanced computed tomography (CT), which revealed multiple low-density lesions in both kidneys and a low density hepatic mass. Laboratory tests showed an extremely high level of serum IgG4. Percutaneous renal and hepatic biopsies showed diffuse infiltration of lymphocytes and IgG4 positive plasma cells with fibrosis in both tissues. Two months after administration of oral prednisolone, both lesions decreased in size on follow-up CT, and the serum creatinine level also improved. No recurrence has been detected for two years with a maintenance dose of prednisolone. PMID- 21628943 TI - Improvement of rheumatoid arthritis and autoimmune hepatitis in a patient treated with the tumor necrosis factor inhibitor, etanercept. AB - We report a case of rheumatoid arthritis (RA) with autoimmune hepatitis (AIH) and Sjogren syndrome (SjS) that was treated with the tumor necrosis factor (TNF) inhibitor, etanercept (ETN). Both RA activity and transaminase levels improved as a result of treatment. Follow-up liver biopsy showed improvement of hepatitis. Although the efficacy of anti-TNF for RA patients with AIH remains controversial, this case suggests that treatment with ETN may result in a favorable clinical course in a certain subset of patients with RA and AIH. PMID- 21628944 TI - Mycobacterium heckeshornense lung infection that was diagnosed as Mycobacterium xenopi disease by DNA-DNA hybridization (DDH). AB - The DNA sequencing analyses of the 16S rRNA gene, rpoB and hsp65 were conducted to characterize six strains that had been identified as Mycobacterium xenopi by DNA-DNA hybridization (DDH) for past ten years in our hospital. The results revealed Mycobacterium heckeshornense infection in one of the six cases. A 47 year-old man, who had been treated for pneumonia, had pulmonary nontuberculous mycobacterial disease. The sputa from the patient were culture positive for mycobacterium in three times. And it was diagnosed as M. xenopiby DDH method. Chest X-ray showed fibrocavitary lesion in right upper lobe was successfully treated with clarithromycin for four weeks. PMID- 21628945 TI - Intussusceptions associated with an ileal duplication cyst. PMID- 21628946 TI - Isolated right-sided pulsus alternans in pulmonary arterial hypertension. PMID- 21628947 TI - Multiple pulmonary artery fistulas causing partial lung edema. PMID- 21628948 TI - Giant mediastinal germ cell tumor. PMID- 21628949 TI - Leriche syndrome detected by 64-MDCT. PMID- 21628950 TI - Multimodality imaging of subcutaneous panniculitis-like T-cell lymphoma. PMID- 21628951 TI - Origin of restenosis after drug-eluting stent implantation in hyperglycemia is inflammatory cells and thrombus. AB - AIMS: The cellular and molecular mechanisms and safety after drug-eluting stent (DES) implantation in diabetic patients are still poorly understood; therefore, in this study, we evaluated the pathologic responses of the sirolimus-eluting stent (SES) or paclitaxel-eluting stent (PES) in a type I diabetes mellitus (DM) rat model. METHODS: The type I DM rat model was manipulated by intra-peritoneal streptozotocin injection. Two weeks later, DES was implanted in the aorta of rats with hyperglycemia or not as a control. Four weeks after DES implantation, the stented aorta was isolated and histomorphometric analysis was performed. RESULTS: On histomorphometric analysis, increased thrombus, inflammatory cell infiltration, and neointimal hyperplasia (NIH) without change of the smooth muscle cell number after DES implantation were observed in DM rats compared with non-DM (NDM) rats. Furthermore, delayed coverage of mature endothelial cells defined as a von Willebrand factor expression and increased immature endothelial cells as a c-kit expression after DES implantation were observed in DM rats compared with NDM rats. Increased fibrin deposition and decreased hyaluronic acid accumulation at NIH after DES implantation were also observed in DM rats compared with NDM rats. CONCLUSIONS: In conclusion, the main mechanism of restenosis after DES implantation under hyperglycemic conditions was initial thrombus with changes of the extracellular matrix rather than SMC proliferation. These results provided a therapeutic clue for the selection of DES and application of combination therapy using anti-thrombotic and anti-inflammatory drugs in diabetic patients. PMID- 21628952 TI - Crucial role of membrane type 1 matrix metalloproteinase (MT1- MMP) in RhoA/Rac1 dependent signaling pathways in thrombin- stimulated endothelial cells. AB - AIM: Thrombin induces vascular responses including the promotion of tissue factor (TF) and plasminogen activator inhibitor-1 (PAI-1) protein expression, which is modulated by small GTPases RhoA and Rac1, Ca(2+) signaling and reactive oxygen species (ROS). Recent studies have shown that membrane type 1 matrix metalloproteinase (MT1-MMP) functions not only as a protease but also as a signaling molecule. In this study, we hypothesized that MT1-MMP may mediate RhoA and Rac1 activation and their downstream events in thrombin-stimulated endothelial cells. METHODS: We used cultured human aortic endothelial cells (HAECs). MT1-MMP was silenced by small interfering RNA (siRNA). RhoA was inhibited by C3 exoenzyme, whereas adenovirus-mediated gene transfection of dominant negative RhoA and Rac1 was used for the inhibition of RhoA and Rac1. RhoA and Rac1 activation was determined by pull-down assays. Intracellular Ca(2+) concentrations ([Ca(2+)](i)) were fluorescently measured by fura-2 assay. NADPH oxidase activity was determined by lucigenin-enhanced chemiluminescence. RESULTS: Inhibition of RhoA attenuated thrombin-triggered [Ca(2+)](i) increase and TF and PAI-1 expression in HAECs, whereas thrombin-triggered ROS generation and TF and PAI-1 expression were blocked by inhibition of Rac1. Silencing of MT1-MMP attenuated thrombin-triggered RhoA and Rac1 activation, resulting in the attenuation of downstream events including Ca(2+) signaling, NADPH oxidase activity, ROS generation, and TF and PAI-1 expression. CONCLUSIONS: The present study shows that MT1-MMP mediates the RhoA/Ca(2+) and Rac1/NADPH oxidase dependent signaling pathways in thrombin-induced vascular responses. PMID- 21628953 TI - Methylmercury inhibits electron transport chain activity and induces cytochrome c release in cerebellum mitochondria. AB - The involvement of oxidative stress has been suggested as a mechanism for toxicity caused by methylmercury (MeHg). One of the major critical sites for oxidative stress is the mitochondria. In this research, to clarify the target site in mitochondria affected by MeHg, the individual activities of the mitochondrial electron transport chain (ETC) (I~IV) were examined in the liver, cerebrum and cerebellum of MeHg-intoxicated rats. In addition, to elucidate the mechanism underlying MeHg toxicity, cytochrome c release, caspase 3 activity and histological study were examined in the cerebrum and cerebellum. The cerebellum was found to be an exclusive tissue in which significant MeHg-induced alterations were observed. The complex II activity in the cerebellum mitochondria significantly decreased after MeHg exposure. Cytochrome c release from mitochondria increased only in the cerebellum by MeHg exposure. However, no significant alterations in caspase 3 activity or histological structure were found in brain tissues. These results suggest that MeHg acts on the constituents of complex II in the cerebellum, and induces mitochondrial dysfunction, leading to a release of cytochrome c from mitochondria. These events were considered to occur at the early stage of MeHg intoxication. PMID- 21628954 TI - Cigarette smoke decreases organic acids levels and population of bifidobacterium in the caecum of rats. AB - Cigarette smoke has been known to affect the development of bowel disease. However it has not been fully elucidated how cigarette smoke has effects on the gut. In this context we evaluated not only caecal levels of organic acids but also populations of micro-flora and pH in caecal contents after exposing rats (n=5) to cigarette smoke for a 4-week in order to investigate whether the gut environment is altered by cigarette smoke or not. After the exposure of cigarette smoke, caecal levels of organic acids such as acetic acid, propionic acid, butyric acid and valeric acid significantly decreased. Additionally the population of Bifidobacterium significantly decreased and the pH significantly elevated. In conclusion cigarette smoke changes caecal levels of certain organic acids, the population of Bifidobacterium and the pH in caecal contents of rats. These results suggest that cigarette smoke may alter the gut environment of rats. PMID- 21628955 TI - Nasal instillation of nanoparticle-rich diesel exhaust particles slightly affects emotional behavior and learning capability in rats. AB - In the present study, in order to reveal novel adverse effects of ultrafine particles (UFP) on the central nervous system, the effects of nanoparticle-rich diesel exhaust particles (NRDEP; count mode diameter, 21.45 nm) on emotional behavior, learning capability and brain neurotransmitter levels were studied in rats by intranasal instillation (iNI). NRDEP (10 and 50 ug/rat) was instilled into 2-week old infant, male rats once a week for 4 weeks. Spontaneous motor activity measured was observed to be inverse to the dose level. In active avoidance tests using a shuttle box, NRDEP-treated animals showed a lower avoidance performance than control animals given air-instillation. The levels of dopamine and its metabolite (DOPAC) in the medial mammillary nucleus of the brain tended to be lower in the NRDEP-treated animals. From these results, although the effects of NRDEP by iNI on the emotionality and the brain neurotransmitter levels were not fully clear, the results obtained by avoidance testing suggested involvement of UFP in learning capability. PMID- 21628956 TI - The mechanism of action of sanguinarine against methicillin-resistant Staphylococcus aureus. AB - Sanguinarine is a benzophenanthridine alkaloid derived from the root of Sanguinaria canadensis. It is known to perform a wide spectrum of biological activities. The aim of this study is to examine the antimicrobial actions of sanguinarine against methicillin-resistant Staphylococcus aureus (MRSA). Sanguinarine antimicrobial activity was assessed by broth dilution method; its mechanism of action was investigated by bacteriolysis, detergent or ATPase inhibitors and transmission electron microscopy were used to monitor the survival characteristics and the changes in bacteria morphology. The activity of sanguinarine against MRSA strains ranged from 3.12 to 6.25 ug/ml, while the minimum inhibitory concentrations of the two reference strains are 3.12 ug/ml and 1.56 ug/ml. The treatment of the cells with sanguinarine induced the release of membrane-bound cell wall autolytic enzymes, which eventually resulted in lysis of the cell. The OD(600s) of the suspensions treated with the combination of Tris (hydroxymethyl) aminomethane and Triton X-100 with sanguinarine were reduced to 40% and 8%, respectively. Transmission electron microsco-py of MRSA treated with sanguinarine showed alterations in septa formation. The predisposition of lysis and the altered morphology seen by transmission electron microscopy suggest that sanguinarine compromises the cytoplasmic membrane. PMID- 21628957 TI - Hepatoprotective effects of vitamin E/selenium against malathion-induced injuries on the antioxidant status and apoptosis-related gene expression in rats. AB - The present study is undertaken to evaluate the protective effect of vitamin E (alpha-tocopherol) and selenium (Se) against malathion (MTN)-induced oxidative stress and hepatic injuries in experimental rats. Male rats were randomly divided into eight groups comprised of 10 rats each. The 1(st) group served as a negative control (C(N)), whereas the 2(nd) was supplemented with a combination of alpha tocopherol (100 mg kg(-1) body weight, b.w.)/Se (0.1 mg kg(-1) bw). The 3(rd), 4(th) and 5(th) groups were respectively administered with increasing doses of MTN equivalent to (1/50 )LD(50) (M(1/50)), (1/25) LD(50) (M(1/25)) and (1/10) LD(50) (M(1/10)), respectively. The 6(th), 7(th) and 8(th) groups were administered the same doses of MTN as in the 3(rd), 4(th) and 5(th) groups with a concomitant supplementation with alpha-tocopherol/Se. Subchronic exposure of rats to MTN for 45 days resulted in statistical dose-dependent decrease in acetylcholinestrase (AChE) activity, increase in oxidative stress marker lipid peroxidation (LPO) and reduction in reduced glutathione (GSH) level. Moreover, the levels of glutathione persoxidase (GPx), superoxide dismutase (SOD) and catalase (CAT) were significantly decline in response to MTN exposure in a dose dependent fashion. Furthermore, histopathological studies of liver in the rats which received MTN exhibited, moderate to severe degenerative and necrotic changes in the hepatocytes. Notably, the administration of alpha-tocopherol/Se protected the liver of rats exposed to MTN as evidenced by the appearance of normal histological structures, significant attenuation of the decline in all antioxidant enzymes tested (i.e. GPx, SOD and CAT), significant recovery in the GSH level and statistical reduction in LPO, as compared to the experimental rat. The effect of alpha-tocopherol/Se supplementation on transcriptional activity of three key stress and apoptosis-related genes (i.e., Tp53, CASP3 and CASP9), in response to MTN exposure in rats, was investigated. Results revealed a significant concentration-dependent up-regulation in the level of expression for the three genes examined, in response to MTN exposure, compared with the control. Interestingly, the supplementation of MTN-treated rats with alpha-tocopherol/Se modulates the observed significant dose-dependent up-regulation in the level of expression for three selected genes, indicative of an interfering role in the signaling transduction process of MTN-mediated poisoning. Taken together, these data suggest that the administration of alpha-tocopherol/Se may partially protect against MTN-induced hepatic oxidative stress and injuries. PMID- 21628958 TI - Lack of chronic toxicity and carcinogenicity of dietary administrated catechin mixture in Wistar Hannover GALAS rats. AB - Chronic toxicity and carcinogenicity of catechin mixture were examined in Wistar Hannover GALAS rats. Administration was in the diet at concentrations of 0, 0.02, 0.3, 1 or 3%. Slight increases in relative liver weight and centrilobular hypertrophy of hepatocytes associated with induction of CYP3A2 were found at the 3% in males of both studies. However, because there were no signs indicative of hepatotoxicity on serum biochemical and histopathological examinations, the changes observed in the liver were regarded as adaptation, and not adverse effects. The slight depressions of body weights at the 3% in females of the chronic toxicity study and in both sexes of the carcinogenicity study were observed. These decreases were because the diet at the highest concentration was frangible and nominal food consumption may not have reflected the actual food consumption resulting in decrease in caloric intake, rather than toxic effects. Thus it was concluded that catechin mixture had no toxicity. In addition, tumor incidences and types were comparable between treated and control groups. Based on the results, the no observed adverse effect levels estimated from the chronic toxicity study were 3% in both sexes equal to 1922.9 in males and 2525.7 mg/kg/day in females. Catechin mixture has no carcinogenic potential in male and female rats. PMID- 21628959 TI - The relationship between CD86 and CD54 protein expression and cytotoxicity following stimulation with contact allergen in THP-1 cells. AB - Contact allergens induce the augmentation of cell surface molecules on and release of cytokines from Langerhans cells (LC) in skin sensitization. THP-1 and U937 cell lines, surrogates of LC, were used as analytical tools of this phenomenon recently. In THP-1 cells, contact allergens are reported to induce the phenotypic alteration including the production of pro-inflammatory cytokines and augmentation of cell surface molecules especially at sub-toxic doses. However, the relationship between phenotypic alteration and cytotoxicity is not clear yet. The purpose of this study is to understand the relationship between the protein expression and cytotoxicity induced by contact allergens. First, we observed that the cytotoxicity induced by contact allergens is caused by both apoptosis and necrosis. Apoptosis was preferentially confirmed in stimulation with contact allergens, but non-allergen sodium lauryl sulfate (SLS) hardly induced apoptosis. Moreover, there was no effect to augmentation of protein expression when apoptosis induction pathways were inhibited. Based on these findings, we proposed that the protein expression and cytotoxicity were controlled independently. Next, oxidative stress was found to be generated by contact allergens at the early phase, and this regulated the protein expression and cytotoxicity at least partially. Finally, the humoral factors from dead cells induced by dinitrochlorobenzene (DNCB) were exposed to fresh THP-1 cells to confirm whether protein expression depended on cytotoxicity. The protein expression was not induced. Altogether, these results suggest that cytotoxicity induced by contact allergens may result in apoptosis and may also be stimulated in parallel with protein expression through an intracellular signal or signals. PMID- 21628960 TI - Effects of spaced feeding on gene expression of hepatic transaminase and gluconeogenic enzymes in rats. AB - Blood alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities are widely used as sensitive markers of liver toxicity. However, these activities are also recognized to be altered by hormonal and nutritional modifications. We investigated the relationships between the activity and gene expression of the hepatic transaminases and the state of hepatic amino acid/glucose/fatty acid metabolism in the ad libitum fed (ALF) and spaced-fed (SF) rats. Acceleration of hepatic gluconeogenesis and fatty acid oxidation was noted in the SF rats. Expression of hepatic clock gene was also altered in the SF rats. Hepatic transaminase activities in the SF rats were higher than those in the ALF rats. These alterations were due to increases in the synthesis of hepatic ALT and AST proteins. In conclusion, the increased transaminase protein synthesis in the liver of the SF rats was considered to be related to the acceleration of hepatic gluconeogenesis under the conditions of spaced feeding. PMID- 21628961 TI - Oral toxicity of pefloxacin, norfloxacin, ofloxacin and ciprofloxacin: comparison of biomechanical and histopathological effects on Achilles tendon in rats. AB - Four fluoroquinolones (pefloxacin, norfloxacin, ofloxacin and ciprofloxacin) were compared according to their biomechanical and histopathological effects on rat Achilles tendon. Wistar rats were divided into one untreated control and four treatment groups in parallel. Pefloxacin mesylate dihydrate (40 mg/kg), norfloxacin (40 mg/kg), ofloxacin (20 mg/kg) and ciprofloxacin (50 mg/kg) were administered by gavage twice daily for three consecutive weeks. 6 weeks after treatment, the test animals were euthanised and Achilles tendon specimens were collected. A computer monitored tensile testing machine was utilised for biomechanical testing. The mean elastic modulus of the control group was significantly higher than that of the norfloxacin and pefloxacin groups (p<0.05 and p<0.01, respectively). The mean yield force (YF) of the control group was significantly higher than those of ciprofloxacin, norfloxacin and pefloxacin groups (p<0.001, p<0.05 and p<0.01, respectively). The mean ultimate tensile force (UTF) of the control group was significantly higher than of the ciprofloxacin, norfloxacin, and pefloxacin groups (p<0.001, p<0.05 and p<0.01, respectively). Hyaline degeneration and fibre disarrangement were observed in the tendons of the ciprofloxacin, pefloxacin, and ofloxacin treated-groups, whereas myxomatous degeneration was observed only in the ciprofloxacin and pefloxacin groups. In conclusion, these findings in our rat model reveal significant deterioration of biomechanical parameters following fluoroquinolone exposure, and indicate significantly higher biomechanical toxicity for ciprofloxacin and pefloxacin. PMID- 21628962 TI - Single administration effects of ethanol on the distribution of white blood cells in rats. AB - Acute single administration effects of ethanol on the distribution of total white blood cells (WBCs), neutrophil, basophil, eosinophil, monocyte and lymphocytes were studied in rats. Acute single administration effects of ethanol on the number of red blood cells (RBCs), hemoglobin concentration and hematocrit were also examined. Male 8-week-old Sprague Dawley rats were randomly divided into the ethanol-administered (ETA) group and the control (CON) group. Two parts of an experiment, 1) 1st experiment : (ethanol dose : 1.0 g/kg body weight), and 2) 2nd experiment : (ethanol dose : 2.0 g/kg body weight) were carried out in rats. The rats were starved to 19:00, and deprived of diet for 12 hr and water for 1 hr before the single administration of ethanol. 1.0 or 2.0 g/kg body weight of ethanol (in 20% (w/w) ethanol) was orally administered to ETA group rats via a stainless stomach tube. In the CON group rats, 0.9% NaCl solution was orally given with the solution volume being equal, in the same manner. Single administration of 1.0 or 2.0 g/kg body weight of ethanol did not change the number of RBCs, hemoglobin concentration and hematocrit. Single administration of 1.0 g/kg body weight of ethanol did not also change the number of WBCs. However, administration of 2.0 g/kg body weight of ethanol increased significantly the number of neutrophil, basophil, monocyte and total WBCs without changing the number of eosinophil and lymphocytes. These results suggest that single administration of 2.0 g/kg body weight of ethanol to rats increased markedly the number of the natural immunity cells without changing the number of acquired cells. PMID- 21628963 TI - Hepatic expression of spermatogenic genes and their transiently remarkable downregulations in Wistar-Kyoto rats in response to lead-nitrate administration: strain-difference in the gene expression patterns. AB - Administration of lead ion (Pb) to rats and mice affects hepatic functions such as the induction of hepatic cell proliferation and upregulation of cholesterol biosynthesis. To identify the genes for which expression changes in response to Pb-administration, we analyzed hepatic gene expression patterns in stroke-prone spontaneously hypertensive rat (SHRSP), its normotensive control, Wistar-Kyoto rat (WKY), and Spraque-Dawley (SD) rat strains, 3, 6, and 12 hr later after single i.v. injection of lead nitrate (LN) at a dose of 100 umol using a DNA microarray technique. The data analysis demonstrated that the expression of a great number of genes was transiently and remarkably downregulated 3 hr after LN injection, and then recovered to control levels only in LN-injected WKY. These normal hepatic expression levels in WKY and SHRSP were much higher than those in SD rats. Furthermore, most of these genes were ones thought to be expressed specifically in the spermatids and/or testes; i.e. genes encoding protamin 1, transition protein 1, and transition protein 2. These findings suggest that the regulation system common to expression of all of these genes could be a target site of Pb-toxic action, at least, in the liver of WKY, and that this system might be similar to the system essential for spermatogenesis, especially spermiogenesis, in the testis. In addition, it appears that clarifying the cause of the difference between the systems of WKY and SHRSP might aid in identifying the pathologic genes in SHRSP. Finally, it will be an important to clarify how the products of the genes related to spermatogenesis, including spermiogenesis, are functional in the livers of WKY and SHRSP. PMID- 21628964 TI - Induction by mercury compounds of metallothioneins in mouse tissues: inorganic mercury accumulation is not a dominant factor for metallothionein induction in the liver. AB - Among the naturally occurring three mercury species, metallic mercury (Hg(0)), inorganic mercury (Hg(II)) and methylmercury (MeHg), Hg(II) is well documented to induce metallothionein (MT) in tissues of injected animals. Although Hg(0) and MeHg are considered to be inert in terms of directly inducing MT, MT can be induced by them after in vivo conversion to Hg(II) in an animal body. In the present study we examined accumulations of inorganic mercury and MT inductions in mouse tissues (brain, liver and kidney) up to 72 hr after treatment by one of three mercury compounds of sub-lethal doses. Exposure to mercury compounds caused significant mercury accumulations in mouse tissues examined, except for the Hg(II)-treated mouse brain. Although MeHg caused the highest total mercury accumulation in all tissues among mercury compounds, the rates of inorganic mercury were less than 10% through the experimental period. MT inductions that depended on the inorganic mercury accumulation were observed in kidney and brain. However, MT induction in the liver could not be accounted for by the inorganic mercury accumulation, but by plasma IL6 levels, marked elevation of which was observed in Hg(II) or MeHg-treated mouse. The present study demonstrated that MT was induced in mouse tissues after each of three mercury compounds, Hg(0), Hg(II) and MeHg, but the induction processes were different among tissues. The induction would occur directly through accumulation of inorganic mercury in brain and kidney, whereas the hepatic MT might be induced secondarily through mercury induced elevation in the plasma cytokines, rather than through mercury accumulation in the tissue. PMID- 21628965 TI - Cyclin D1/cdk4, estrogen receptors alpha and beta, in N-methyl-N'-nitro-N nitrosoguanidine-induced rat gastric carcinogenesis: immunohistochemical study. AB - Hyperproliferative cell growth due to cyclin D1/cdk4, marker of cellular proliferation, is considered to be regulated by the expression of estrogen receptors (ERs). We investigated the immunohistochemical expression of cyclin D1/cdk4 and ERs in N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced rat gastric carcinogenesis. The gastric cancer incidence and expression of cyclin D1/ckd4 in gastric carcinogenesis were significantly higher in males than females. Although the ERalpha expression index was similar in both sexes, the ERbeta expression in preneoplastic hyperplastic lesions as well as gastric cancers was significantly higher in females than in males. The present study revealed a gender difference in MNNG-induced rat gastric carcinogenesis that seemed to involve the sex difference in cyclin D1/cdk4 expression, and ERbeta expression became evident at the preneoplastic promotion stage in gastric carcinogenesis. PMID- 21628966 TI - Differential effects of single-walled carbon nanotubes on cell viability of human lung and pharynx carcinoma cell lines. AB - Carbon nanotubes (CNTs) are attracting significant attention as a novel material for future innovations. Many in vitro studies have assessed the cytotoxicity of CNTs, but the effects of CNTs differ depending on the cell lines and the synthetic method adopted for fabricating CNTs. In the present study, the differential effects of single-walled CNTs (SWCNTs) on the cell viability of A549 cells from human lung carcinomas and FaDu cells from human head and neck carcinomas were investigated. The SWCNTs used in the present study were synthesized with nickel and yttrium (SO-SWCNTs), and iron (FH-P-SWCNTs) as catalysts. Cell viability was evaluated on the basis of cell-membrane biomass, adenosine triphosphate (ATP) content, and intracellular metabolic capacity. After 24-hr exposure of A549 and FaDu cells to 1.0 mg/ml SO-SWCNTs, the cell-membrane biomass of A549 cells decreased to 43% as compared to the control cells, whereas that of FaDu cells remained over 90%. After 24-hr exposure of A549 and FaDu cells to 1.0 mg/ml SO-SWCNT, the intracellular metabolic capacity decreased to 24% and 37%, respectively, and the ATP content decreased to 40% and 54%, respectively. SWCNTs had a greater impact on the viability values of A549 cells than on those of FaDu cells. In addition, cells exposed to FH-P-SWCNTs exhibited a higher viability than those exposed to SO-SWCNTs. Caspase 3/7 activity was not increased at maximum concentration of 1.0 mg/ml SO-SWCNTs. It was surmised that sensitivity to SWCNTs differs among the 2 cell lines; additionally, SWCNT characteristics may produce different effects on these cell lines. PMID- 21628968 TI - [Patients' consent and drug safety of investigational products and approved drugs]. PMID- 21628967 TI - Gene expression profiling using DNA microarray analysis of the cerebellum of mice treated with methylmercury. AB - To elucidate the molecular mechanism involved in methylmercury-induced cerebellar disorder, we performed DNA microarray analysis of the cerebellum of methylmercury treated mice. The expression levels of 21 genes were elevated 2-fold or higher in response to methylmercury, including many genes encoding proteins involved in inflammatory reactions associated with chemokines. The expression levels of 11 genes were reduced by half or more in response to methylmercury. PMID- 21628969 TI - [Pharmacists contribute to the safety of medicines]. AB - Pharmacists have a professional obligation in medicine. As a member of a medical team to provide a safe medication, it is important that they be involved with drug safety in mind. Currently a pharmacist in a business operating with a focus on traditional dispensing and drug administration also functions with a focus on patient care and the provision of drug information. It is important that a pharmacist has a risk management approach to medicine. Together with the institutions responsible for drug safety, he must be cognizant of published reports to prevent serious adverse drug reactions as well as taking part in post marketing surveillance. Management of safety information for high-risk pharmaceutical drugs, also falls to the pharmacist. Thus he must have knowledge of the skill required for the job of each member of the health care team. Recent newly added responsibilities are: advice in planning patient treatment, checking vital signs and the prescription brought to him to fill. In short a pharmacist working in the medical field must, above all, respect and assure safety to the patients he serves. PMID- 21628970 TI - [Scientific drug safety information for patients' consent]. AB - One of the important roles of pharmacists is to continue their contributions to new drug discovery and development. However, it seems to be very difficult to obtain patient satisfaction with new drugs. Because new medicines have both benefit and risk, there should be many systems to maximize the safety and efficacy of the drugs. In clinical trials, the rights, safety and welfare of human subjects under the investigator's care must be protected. Good Clinical Practice is a harmonized ICH-guideline, and the safety information of an investigational product is explained to patients who voluntarily enter the clinical trials. Since safety information about investigational products is still limited, subjects are informed about the results of animal experiments and those of finished clinical trials. The sponsor of clinical trials should be responsible for the on-going safety evaluation of the investigational products. When additional safety information is collected in the clinical trials, the written informed consent form should be appropriately revised. During the review process, quality, safety and efficacy of new drugs are evaluated and judged based on the scientific risk-benefit balance. The safety information collected in clinical trials is reflected in the decision-making process written in the review reports. All-case investigation should be also performed until data from a certain number of patients has been accumulated in order to collect early safety and efficacy data. Important messages written in review reports for drug safety and patient consent are explained. Risk communication will improve the application of patients' consent for new drugs. PMID- 21628971 TI - [Demands and expectations for drug companies]. AB - We need the surroundings in which we can conduct sciences-based safety measurements and continue to do scientific approach in order to ensure the safety of drugs and put patients and the nation at ease. It is important that we actively conduct clinical trials to evaluate the efficacy and safety of drugs, and supply the results to medical scene quickly. Japanese government draws up some plans to activate clinical trials, and intends to improve national health and promote ability of research and development of drugs. Since activation of clinical trials is important for progression of medicine too, drug companies are also expected to take part in it positively. PMID- 21628972 TI - [Drug safety--from patients' perspective]. AB - Patients expect drugs are 100% effective and safe. Unfortunately, however, most drugs are not. Continuous efforts by healthcare professionals and industry should be made to maximize efficacy and safety. Here, four challenges are shown from a viewpoint of laypersons. 1) Develop better drugs: Continuous efforts to develop drugs for 'neglected' diseases should be enhanced to meet unmet medical needs. 2) Deliver right drugs: Medication errors caused by similar names and shapes have been repeatedly reported. Communication with patients and their families may be helpful to decrease errors. 3) Improve the quality of drug information: How health professionals provide drug information to patients should be routinely monitored to improve the quality. Rephrasing to plain expressions may sometimes be useful for better communication. 4) Promote personalized medicine: Each patient wants to know whether this drug would work to him/herself as well as statistical data. Pharmacogenomics and pharmacokinetics/pharmacodynamics (PK/PD) research should be encouraged in order to develop personalized medicine. PMID- 21628973 TI - [Post-marketing drug safety measures for the attainment of safer and more effective use of drug]. AB - In contrast with the 20th century's dramatic improvements in the direct and/or hazardous toxicity of drugs, indirect toxicity and/or long-term safety concerns such as relation of cancer risk and TNF-alpha receptor blockers have caused significant complexity in post-marketing surveillance (PMS) scenery. The post marketing phase of drugs and their safety measures now appear to be much more complicated and heavier than decades ago. The spontaneous adverse drug reaction (ADR) reporting system which has been one of the main pillars of PMS measures for almost 50 years may have to be reviewed in terms of its effectiveness, and may need augmentation from medical data bases. Only a pharmaco-epidemiological analysis and integration of the output with a conventional spontaneous reporting approach offers a chance to satisfy the current complex safety issues. Today's tendency toward practical saturation at medical/pharmaceutical frontiers, by regulatory authorities and safety divisions of pharmaceutical companies with ever increasing day-to-day safety information can also be pointed out. Such phenomena may actually reduce the productivity of safety measures and also jeopardize the maintenance of an acceptable risk/benefit drug ratio. To alleviate these potential negative implications, establishment of a consortium to act as a sentinel that would gather up-to-date and essential safety information, including epidemiological data, from all sources and provide it plus recommendations to all stakeholders can be suggested. Through such activities, we could expect significant improvement of drug safety measures in post-marketing phase which would effectively cover not only new drugs but also generic and bio-simulated drugs. PMID- 21628974 TI - [Challenging the development of new anti-diabetic medicine--approaching from educational, clinical, and research fields]. PMID- 21628975 TI - [Learn and relation, basic knowledge of diabetes--approach from an educational front]. AB - A six-year program started in 2006, and Computer Based Testing (CBT), Objective Structured Clinical Examination (OSCE) and long-term practical training were introduced into the pharmacy education. As a member of the team medical care, we participate in medical care positively, and a pharmacist now wanted is who could be able to communication well with the medical worker and the patient has. We have the results of 32 years as a cramming school for the pharmacist national examination candidate. We have life study center, a hospital, a pharmacy, and the publishing division. Firstly we taking part in the education, learn the present conditions, the clinical cases, and stream down it to a student, and it is necessary to send it in the future of a pharmacy and the pharmacist. Therefore we carry out studying society of the prescription, an experience of the compounding of medicines training, participation to the life study. At this symposium, we give a lecture on diabetes based on the situation of pharmacy education, the clinical spot, medical supplies development. We introduce condition of a patient physiology of the diabetes, the difference of the model, a symptom, inspection, efficacy of the medicine, a side effect in that. Thus, we want to propose some technical skills for relating these clinical conditions in the future education. PMID- 21628976 TI - [Present condition of diabetes and its treatment--from the perspective of a hospital pharmacist]. AB - In Japan, the number of diabetic patients has increased steadily. According to a 2007 report by the Ministry of Health, Labour and Welfare, the number of diabetic patients and its reserves is estimated to be 22 million, or a fifth of adults. On a global scale, the number of deaths due to diabetes is almost equivalent to that due to AIDS. Therefore, in 2006, WHO designated November 14 as World Diabetes Day and began a worldwide campaign to increase people's awareness of diabetes. Diabetic patients usually fluctuate between good and bad data of SMBG (self monitoring of blood glucose). However, the main goal of diabetes treatment is to not only keep good data of SMBG but also prevent diabetes-related complications from arising, because such complications have a major impact on patients' QOL and medical costs. At medical institutions, CDEJs (Certified Diabetes Educator of Japan, a qualification awarded to health care professionals such as nurses, dietitians, pharmacists, laboratory technicians, and physiotherapists) provide information to diabetic patients in order to increase their knowledge of the disease; such professionals also assist with patients' medical treatments. I would like to discuss frequently used diabetes medications, the latest treatments for diabetes, the problems faced by diabetic patients, and so on, from the perspective of a hospital pharmacist. PMID- 21628977 TI - [Search for antidiabetic constituents of medicinal food]. AB - Many foods are known to have not only nutritive and taste values but also medicinal effects. In Chinese traditional medicine, the treatment using medicinal foods has been recommended highly. Recently, we examined the effects of the extract and constituents of several medicinal foods on experimental models of diabetes. In this paper, we focus on the bioactive constituents of four medicinal foods, namely the antidiabetic constituents from 1) the roots, stems and leaves of Salacia plants, 2) the male flowers of Borassus flabellifer, 3) the flower buds of Camellia sinensis, 4) the processed leaves of Hydrangea macrophylla var. thunbergii (Hydrangeae Dulcis Folium). PMID- 21628978 TI - [Challenge of creating single-agents for the treatment of type 1 and 2 diabetes by targeting retinoid X receptor]. AB - It might be seen as reckless to challenge to create single-agents for the treatment of both type 1 diabetes caused by the destruction of the Langerhans beta cells in pancreas by excessive autoimmunity, and type 2 diabetes caused by the obesity. However, we hypothesized that retinoid X receptor (RXR) agonists, which are researched for the treatment of type 2 diabetes, will also be useful like metformin, which shows insulin-sparing effect in type 1 diabetes. This is because PPARgamma/RXR is known to be a target of thiazolidinediones (TZDs), which are used for the treatment of insulin resistance, LXR/RXR is reported to be involved in glucose/lipid metabolism, and these heterodimers can be activated by RXR agonists alone (permissive mechanism). However, repeated administration of RXR agonists can elevate blood triglyceride and induce hypothyroidism. In this study, we performed systematic conversion of the alkoxy side chain of 5a (6 [ethyl-(3-isopropoxy-4-isopropylphenyl)amino] nicotinic acid: NEt-3IP) and evaluated the RXR-, PPAR/RXR- and LXR/RXR-agonist activities of the products. The cyclopropylmethoxy analog (5c) showed similar RXR- and LXR/RXR-agonistic activities to the benzyloxy analog (5i) and n-propoxy analog (5k), but exhibited more potent PPAR/RXR-agonistic activity than 5i or 5k. Differential modulation of RXR heterodimer-activating ability by conversion of the alkoxy group located in the lipophilic domain of the RXR-agonist common structure is expected to be a useful approach in the design of new RXR agonists for the treatment of hyperlipidemia. PMID- 21628979 TI - [Challenge of studies on the development of new Zn complexes to treat diabetes mellitus]. AB - In recent years, people all over the world have suffered from various diseases such as cancer, myocardial infarction, osteoporosis, hypertension, and diabetes mellitus (DM). Especially, DM, well-known as one of lifestyle-related diseases, has been regarded as a serious problem, because it is difficult to fully recover. The number of patients suffering from DM in 2007 was reported to be approximately 200 million people worldwide. However, insulin preparations and synthetic therapeutics, which are clinically used treatment of DM, have been associated with problems such as physical and mental pain due to daily injections and certain severe side effects, respectively. Zn, which is an essential trace element in animals and humans and plays an important role in maintenance of their lives, has been indicated to exhibit insulin-like activity. Since the finding of insulin-like effects of Zn, several Zn complexes have been proposed as a new type of anti-diabetic therapeutics which is differ from existing medicines. In this symposium, we introduce the anti-diabetic effect, complication relieving effect, and action mechanism of bis(2-mercaptopyridine-N-oxidato)Zn complex with Zn(S(2)O(2)) coordination mode. PMID- 21628980 TI - [Education of graduate school of pharmaceutical sciences in new generation: introduction]. PMID- 21628981 TI - [Education for six-years graduate school of pharmaceutical sciences: a case of Tohoku University]. AB - We would like to introduce our new project to develop a course program for the 4 year system of graduate school of pharmacy Tohoku University after the 6-year program of the college of pharmacy. New course program designed will be two fields of 'cancer chemotherapy' and 'life-style related diseases' for clinical specialists and scientists of pharmacy. Our vision through this new program is to educate and produce the preeminent personnel being responsible for the safety and effective pharmaceutical care service in the clinical settings, with having the practical professional knowledge, skill and highly research ability, such as doing intervention and recommendation against a prescription planning process with an appropriate drug use. The personnel educated using our program will be called 'a next generation type pharmacist (or a Japanese-style pharmacist practitioner)'. Our program is a joint project with the graduate school of medicine and the university hospital, Tohoku University. The College of Pharmacy, Tohoku University placed two new laboratories and employed six faculties who are registered pharmacists in order to achieve above, and they also hold the post of pharmacist of the university hospital. And graduate students who are registered pharmacists also work in the university hospital now. PMID- 21628982 TI - [Expectations for the education of graduate school of pharmaceutical sciences from the viewpoint of discovery and development of new drugs]. AB - For the purpose of the development of new drugs for incurable diseases, many students enter graduate school of pharmaceutical sciences every year. At first, I expect education to let it develop more and spread without forgetting this will. Recently, withdrawals from Japan of the research institutes of the foreign affiliated pharmaceutical companies have occurred successively. It is pointed out that there is it for the study about the biomedical research that is the next step of fundamental researches having been weak. I expect the immediate construction of the cluster, which consists of pharmaceutical companies and graduate schools of pharmaceutical sciences. Time of ten several years and a cost of one hundred billion yen are necessary for the research and development of new drug. The success probability is low, besides. Many trials are accomplished to raise the success probability. The one is introduction of the project system. The best members are gathered from the fields such as medicinal chemistry, molecular biology, biochemistry, pharmacology, pharmacokinetics, pharmaceutics and toxicological sciences, etc. The project system is a system enforcing go or stop by own judgment, an authority and the responsibility of the purpose are given. It is necessary for the project leader to have great knowledge and the abilities to hold lively discussion. It is a researcher from graduate school of pharmaceutical sciences that is the most suited to be as a project leader. I expect to upbring education from the time when a leader is young. PMID- 21628983 TI - [Education in graduate school of pharmaceutical sciences]. AB - The report of the Council for the improvement in the education of pharmaceutical sciences and the recommendation of the Central Council for Education indicate that the 6-year education is required to develop pharmacists with high qualities as medical staff. Each college of pharmacy started the education and practical training based on the model core-curriculum with the original program. On the other hand, to develop a scientist for the development of novel medicines, 4-year education program is also required. Under these new education systems, what we should do in the education in the graduate school of pharmacy and pharmaceutical sciences has been discussed. Recently, the first report about the purpose and the strategy in the graduate school in the new generation was submitted. Here, I will comment on the details of this report. PMID- 21628984 TI - [Distribution and accumulation of antibiotics in cells and tissues and toxicity studies by immunocytochemistry]. AB - No true immunocytochemistry (ICC) for drugs nor its application to pharmacokinetic studies is available. Recently, our studies have shown that ICC for drugs is extremely useful for such studies by utilizing easy and safe techniques, and gives direct evidence of drug localization. We have therefore developed antibodies and a series of pretreatment conditions for the immunodetection of drugs and have localized sites of drug uptake or accumulation in several tissues of rats following the administration of drugs. This review describes preparation of anti-drug antibody, specificity of antibody, fixation of drug in situ in rat tissues and cells, treatment of paraffin section specimens prior to immunoreaction, precision, and their application to a variety of types of antibiotics anti-cancer anthracyclines daunorubicin, doxorubicin, and epirubicin, bleomycin analog peplomycin, antimicrobial agents gentamicin, and amoxicillin. ICC for the anti-cancer anthracyclines demonstrated that the drug accumulates in a characteristic pattern in the heart, liver, kidney, gastrointestinal tract, and hair follicles, which represent the sites targeted by the drug toxicity. Some, but not all, of these drug accumulations are associated with the induction of apoptosis. It was also noted that there are striking differences in accumulation among the anthracyclines in rat tissues, maybe contributing the mechanisms of the differences in anti-tumor activities of the anthracyclines. Both ICCs for gentamicin and peplomycin identified characteristic necrotic-like cells in the specific sites of the kidney, suggesting the sites are readily affected by some chemotherapeutic agents. ICC for amoxicillin demonstrated that the sites of the drug accumulation in small intestine, liver and kidney are closely correlated with the specific sites in which certain transporter systems for penicillin occur. Thus, an ICC method is a potential new tool for pharmacokinetic studies of wide variety types of drugs containing a primary amino group(s) in their molecules. PMID- 21628985 TI - Sasa quelpaertensis phenylpropanoid derivative suppresses lipopolysaccharide induced nitric oxide synthase and cyclo-oxygenase-2 expressions in RAW 264.7 cells. AB - 3-O-p-Coumaroyl-1-(4-hydroxy-3,5-dimethoxyphenyl)-1-O-beta-D gulcopyranosylpropanol (ESQ10) is a naturally occurring phenylpropanoid derivative isolated from Sasa quelpaertensis (Gramineae). In the present study, we discovered that ESQ10 inhibits nitric oxide (NO) and prostaglandin E(2) (PGE(2)) production in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. ESQ10 attenuated LPS-induced synthesis of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in parallel and inhibited LPS-induced interleukin-6 production, as determined by an enzyme-linked immunosorbent assay in the macrophages. The mechanism of the antiinflammatory action of ESQ10, i.e., suppression of nuclear factor (NF)-kappaB and mitogen-activated protein kinase activation, has been documented. However, ESQ10 could not influence LPS-mediated IkappaB-alpha degradation and extracellular signal-regulated kinase/c-Jun amino terminal kinase phosphorylation at concentrations of up to 373 uM. To test the potential application of ESQ10 as a topical material, we also conducted a 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay on human HaCaT keratinocytes as well as human dermal fibroblast cells. In this assay, ESQ10 did not induce cytotoxicity. Taken together, the results suggest that ESQ10 may be considered an antiinflammatory candidate for treating inflammatory and skin diseases. PMID- 21628986 TI - Current status and future directions for clinical trials pharmacy. AB - This survey was conducted to investigate the current status and suggest future directions for clinical trials pharmacy in Korea. A 32-item survey was distributed to 96 hospital pharmacies conducting clinical trials. The questionnaire was designed to elicit information on the following: (1) general clinical trial status of each hospital, (2) Investigational Drug Service (IDS) performance status. The response rate was 59.4% and 61.8% of the respondents carried out all the regulatory IDSs. Independent of the IDS's performance, the respondent started to feel the need for reinforcement in human resources when the number of active studies crossed 35. Analyzing the workload based on the subjective need for reinforcement in CTP, the overall CTP work execution rate was significantly higher in the 'needs reinforcement' group (p<0.05), even though this group had a 2.3 times higher workload than that in the 'current number of CTPs is adequate' group. The 'needs reinforcement' group performed more efficiently with a better understanding of IDSs, even though the group was having difficulties due to the shortage of CTPs. The lower execution rate in the 'current number of CTPs is adequate' group can be assumed to be due to the lack of understanding of the scope of IDSs. The work of CTPs should evolve into one of the specialized hospital pharmacists' roles, and a trial institution should designate at least one CTP per 40 protocols. Furthermore, the education for CTPs should be diversified into basic course and advanced course based on the IDS's performance of each hospital. PMID- 21628987 TI - Identification of dimethoate-containing water using partitioned dispersive liquid liquid microextraction coupled with near-infrared spectroscopy. AB - A simple, rapid and efficient extraction procedure, partitioned dispersive liquid liquid microextraction, has been developed in combination with near-infrared spectroscopy for the extraction and discrimination of dimethoate from aqueous samples. For this technique, the appropriate mixture of extraction solvent (CCl(4)) and disperser solvent (THF) was utilized. Partial least squares discriminant analysis was applied to build the model with several pre-process methods over the wavenumber regions between 7100 cm(-1) to 7300 cm(-1). The best model gave satisfactory classification accuracy, 98.6% for calibration set (n=74) and 97.6% for prediction set (n=42), using preprocessing of standard normal variate followed by Savitzky-Golay first derivative. The method was successfully applied to bottled water, tap water, lake water and farm water samples. The results demonstrated the possibility of near-infrared spectroscopy after partitioned dispersive liquid-liquid microextraction for the identification of water contaminated by dimethoate. PMID- 21628988 TI - [In vitro evaluation for corneal damages by anti-glaucoma combination eye drops using human corneal epithelial cell (HCE-T)]. AB - The combination of anti-glaucoma eye drops is frequently used in clinical treatment, and it is known that the combination can cause corneal damage. Recently, an anti-glaucoma combination eye drops is developed, and the treatment by the combination eye drops is expected to enhance quality of life. However, effects of the combination eye drops on corneal epithelial cell damage have not been clarified. In this study, we investigated the corneal epithelial cell damage of commercially available anti-glaucoma combination eye drops, such as Xalacom(r) (latanoprost/timolol maleate combination eye drops), Duotrav(r) (travoprost/timolol maleate combination eye drops) and Cosopt(r) (dorzolamide hydrochloride/timolol maleate combination eye drops) using the human corneal epithelial cell (HCE-T). The cytotoxicity in Xalacom(r) was higher than that in Xalatan(r) (eye drops containing latanoprost) and Timoptol(r) (eye drops containing timolol maleate), and the benzalkonium chloride (BAC) and timolol maleate were related to cytotoxicity in Xalacom(r). The cytotoxicity in Duotrav(r) and Cosopt(r) was lower than that in Timoptol(r). The Duotrav(r) is preserved with a non-BAC system (POLYQUAD, polidronium chloride). Therefore, it was suggested that the POLYQUAD related to the low cytotoxicity in Duotrav(r). On the other hand, the D-mannitol reduced the cytotoxicity by BAC in this study. This result suggested that the cytotoxicity in Cosopt(r) was reduced by D mannitol. The Duotrav(r) and Cosopt(r) may be less damaging to the ocular surface of glaucoma patients receiving long-term eye drop therapy in compared with the combination of anti-glaucoma eye drops. PMID- 21628989 TI - Induction of differentiation by panaxydol in human hepatocarcinoma SMMC-7721 cells via cAMP and MAP kinase dependent mechanism. AB - Panaxydol (PND) is one of the main non-peptidyl small molecules isolated from the lipophilic fractions of Panax notoginseng. The present study was carried out to demonstrate the potential effects of panaxydol on the induction of differentiation of human liver carcinoma cell lines SMMC-7721. Cell viability was evaluated by MTT method and Trypan blue exclusion assay respectively. The changes of morphology were detected by transmission electron microscope. Inhibitors were applied to detect the signaling pathway of differentiation. The level of intracellular cyclic AMP was determined by radioimmunoassay. The expression of p ERK, Id1, and p21 were determined by Western blot. We found that panaxydol inhibit the proliferation of SMMC-7721 cells and caused the morphology and ultrastructure changes of SMMC-7721. Moreover, panaxydol dose-dependently increased the secretion of albumin and alkaline phosphatase activity, and decreased the secretion of AFP correspondingly. These changes of differentiation markers in SMMC-7721 can be reversed by the protein kinase A inhibitor RpcAMPS and by MAP kinase kinase 1/2 inhibitor U0126 or sorafenib. Intracellular cAMP was elevated by panaxydol in SMMC-7721 cells. Panaxydol dose-dependently decreased the expression of regulatory factors Id1 and increased the protein levels of p21 and p-ERK1/2 correspondingly. It suggested panaxydol might be of value for further exploration as a potential anti-cancer agent via cAMP and MAP kinase dependent mechanism. PMID- 21628990 TI - The role of promoter methylation in Epstein-Barr virus (EBV) microRNA expression in EBV-infected B cell lines. AB - Epstein-Barr virus (EBV) microRNAs (miRNAs) are expressed in EBV-associated tumors and cell lines, but the regulation mechanism of their expression is unclear yet. We investigated whether the expression of EBV miRNAs is epigenetically regulated in EBV-infected B cell lines. The expression of BART miRNAs was inversely related with the methylation level of the BART promoter at both steady-state and following 5-aza-2'-deoxycytidine treatment of the cells. The expression of BHRF1 miRNAs also became detectable with the demethylation of Cp/Wp in latency I EBV-infected cell lines. Furthermore, in vitro methylation of the BART and Cp promoters reduced the promoter-driven transactivation. In contrast, tricostatin A had little effect on the expression of EBV miRNA expression as well as on the BART and Cp/Wp promoters. Our results suggest that promoter methylation, but not histone acetylation, plays a role in regulation of the EBV miRNA expression in EBV-infected B cell lines. PMID- 21628991 TI - Genome-wide scan of granular corneal dystrophy, type II: confirmation of chromosome 5q31 and identification of new co-segregated loci on chromosome 3q26.3. AB - Granular corneal dystrophy, type II (CGD2; Avellino corneal dystrophy) is the most common corneal dystrophy among Koreans, but its pathophysiology is still poorly understood. Many reports showed that even though the causative mutation is the same TGFBI R124H mutation, there are severe and mild phenotypes of the corneal dystrophy. We also observed the phenotype differences in our samples. For this reason, we focused our effort on the identification of unknown genetic factor related to phenotype variation. A total 551 individuals from 59 families were genotyped with SNP chip and used in genome-wide linkage analysis. From single-point linkage analyses, we confirmed the known 5q31 region for TGFBI gene, and selected novel nine candidate loci for CGD2. In simulation analysis, the only 3q26.3 region including neuroligin 1 gene (NLGN1) was supported by empirical statistic significance. To investigate the effect of genetic heterogeneity in linkage analysis, we classified CGD2 families into two subgroups. Although we could not find a significant evidence for correlation between the 3q26.3 region and CGD2 phenotypes, this first genome-wide analysis with CGD2 families in Korea has a very important value for offering insights in genetics of CGD2. In addition, the co-segregating loci with CGD2 including 3q26.3 would be a good target for further study to understand the pathophysiology of CGD2. PMID- 21628993 TI - Epigenetics in disease: leader or follower? AB - Epigenetic silencing is a pervasive mode of gene regulation in multicellular eukaryotes: stable differentiation of somatic cell types requires the maintenance of subsets of genes in an active or silent state. The variety of molecules involved, and the requirement for active maintenance of epigenetic states, creates the potential for errors on a large scale. When epigenetic errors - or epimutations - activate or inactivate a critical gene, they may cause disease. An epimutation that occurs in the germline or early embryo can affect all, or most, of the soma and phenocopy genetic disease. But the stochastic and reversible nature of epigenetic phenomena predicts that epimutations are likely to be mosaic and inherited in a nonmendelian manner; epigenetic diseases will thus rarely behave in the comfortably predictable manner of genetic diseases but will display variable expressivity and complex patterns of inheritance. Much phenotypic variation and common disease might be explained by epigenetic variation and aberration. The known examples of true epigenetic disease are at present limited, but this may reflect only the difficulty in distinguishing causal epigenetic aberrations from those that are merely consequences of disease, a challenge further extended by the impact of environmental agents on epigenetic mechanisms. The rapidly developing molecular characterization of epigenomes, and the new ability to survey epigenetic marks on whole genomes, may answer many questions about the causal role of epigenetics in disease; these answers have the potential to transform our understanding of human disease. PMID- 21628992 TI - p41-Arc, a regulatory subunit of Arp2/3 complex, can induce premature senescence in the absence of p53 and Rb. AB - Cellular senescence is a tumor-suppressive process instigated by proliferation in the absence of telomere replication, by cellular stresses such as oncogene activation, or by activation of the tumor suppressor proteins, such as Rb or p53. This process is characterized by an irreversible cell cycle exit, a unique morphology, and expression of senescence-associated-beta-galactosidase (SA-beta gal). Despite the potential biological importance of cellular senescence, little is known of the mechanisms leading to the senescent phenotype. p41-Arc has been known to be a putative regulatory component of the mammalian Arp2/3 complex, which is required for the formation of branched networks of actin filaments at the cell cortex. In this study, we demonstrate that p41-Arc can induce senescent phenotypes when it is overexpressed in human tumor cell line, SaOs-2, which is deficient in p53 and Rb tumor suppressor genes, implying that p41 can induce senescence in a p53-independent way. p41-Arc overexpression causes a change in actin filaments, accumulating actin filaments in nuclei. Therefore, these results imply that a change in actin filament can trigger an intrinsic senescence program in the absence of p53 and Rb tumor suppressor genes. PMID- 21628994 TI - Prion protein function and the disturbance of early embryonic development in zebrafish. AB - Transmissible Spongiform Encephalopathies (TSE) or prion diseases are a threat to food safety and to human and animal health. The molecular mechanisms responsible for prion diseases share similarities with a wider group of neurodegenerative disorders including Alzheimer disease and Parkinson disease and the central pathological event is a disturbance of protein folding of a normal cellular protein that is eventually accompanied by neuronal cell death and the death of the host. Prion protein (PrP) is a constituent of most normal mammalian cells and its presence is essential in the pathogenesis of TSE. However, the function of this normal cellular protein remains unclear. The prevention of PRNP gene expression in mammalian species has been undramatic, implying a functional redundancy. Yet PrP is conserved from mammals to fish. Recent studies of PrP in zebrafish have yielded novel findings showing that PrP has essential roles in early embryonic development. The amenability of zebrafish to global technologies has generated data indicating the existence of "anchorless" splice variants of PrP in the early embryo. This paper will discuss the possibility that the experimentalist's view of PrP functions might be clearer at a greater phylogenetic distance. PMID- 21628995 TI - Sea urchin embryos as a model system for studying autophagy induced by cadmium stress. AB - It is well known that sea urchin embryos are able to activate different defense strategies against stress. We previously demonstrated that cadmium treatment triggers the accumulation of metal in embryonic cells and the activation of defense systems depending on concentration and exposure time, through the synthesis of heat shock proteins and/or the initiation of apoptosis. Here we show that Paracentrotus lividus embryos exposed to Cd adopt autophagy as an additional stratagem to safeguard the developmental program. At present, there are no data focusing on the role of this process in embryo development of marine organisms. PMID- 21628996 TI - In vivo optical imaging of motor neuron autophagy in a mouse model of amyotrophic lateral sclerosis. AB - Autophagy is involved in the pathological process of motor neuron death in amyotrophic lateral sclerosis (ALS). We have generated a novel double transgenic (DTg) mouse line by mating a green fluorescent protein (GFP)-fused microtubule associated protein 1 light chain 3 (LC3) transgenic (LC3-Tg) mouse and a G93A mutant human Cu/Zn superoxide dismutase (mSOD1) transgenic (mSOD1-Tg) mouse. In vivo imaging of autophagy with these novel DTg mice was conducted at 10 (presymptomatic), 17 (early symptomatic) and 19 (late symptomatic) weeks of age. Fluorescence imaging analysis revealed a strong fluorescent signal in vivo over the T3-S1 level at 17 and 19 weeks of age only in the DTg mice. Ex vivo autophagy imaging of spinal cord sections (20 MUm) also showed a progressive increase of the fluorescence signal from 17 to 19 weeks in DTg mice in the anterior horn at the L4-5 level, and the fluorescence signals were clearly observed in the gray matter of the spinal cord with a progressive increase of the signal and decreases in large motor neurons. Protein gel blot analysis revealed maximum LC3-I and LC3 II expressions at 19 weeks, consistent with the results from the in vivo autophagy imaging experiment. This method could also be applied as a unique tool for clarifying the role of autophagy, and to monitor the pathologic processes involving autophagy not only in ALS, but also other neurological diseases. PMID- 21628998 TI - How does timing, duration and severity of heat stress influence pollen-pistil interactions in angiosperms? AB - Reproductive development in sexual plants is substantially more sensitive to high temperature stress than vegetative development, resulting in negative implications for food and fiber production under the moderate temperature increases projected to result from global climate change. High temperature exposure either during early pollen development or during the progamic phase of pollen development will negatively impact pollen performance and reproductive output; both phases of pollen development are considered exceptionally sensitive to moderate heat stress. However, moderately elevated temperatures either before or during the progamic phase can limit fertilization by negatively impacting important pollen pistil interactions required for successful pollen tube growth toward the ovules. This minireview identifies the impacts of heat stress on pollen-pistil interactions and sexual reproduction in angiosperms. A special emphasis is placed on the biochemical response of the pistil to moderately high temperature and the resultant influence on in vivo pollen performance and fertilization. PMID- 21628997 TI - Signal processing by protein tyrosine phosphorylation in plants. AB - Protein phosphorylation is a reversible post-translational modification controlling many biological processes. Most phosphorylation occurs on serine and threonine, and to a less extend on tyrosine (Tyr). In animals, Tyr phosphorylation is crucial for the regulation of many responses such as growth or differentiation. Only recently with the development of mass spectrometry, it has been reported that Tyr phosphorylation is as important in plants as in animals. The genes encoding protein Tyr kinases and protein Tyr phosphatases have been identified in the Arabidopsis thaliana genome. Putative substrates of these enzymes, and thus Tyr-phosphorylated proteins have been reported by proteomic studies based on accurate mass spectrometry analysis of the phosphopeptides and phosphoproteins. Biochemical approaches, pharmacology and genetic manipulations have indicated that responses to stress and developmental processes involve changes in protein Tyr phosphorylation. The aim of this review is to present an update on Tyr phosphorylation in plants in order to better assess the role of this post-translational modification in plant physiology. PMID- 21629000 TI - The President's Health Care Reform Plan and Older Adults: Expanded Research Agenda for Geriatric Psychiatry. PMID- 21628999 TI - Differential regulation and localization of carboxypeptidase D and carboxypeptidase E in human and mouse beta-cells. AB - Hyperglycemia can result from a relative or absolute lack of functional insulin secreted by the pancreatic beta-cells. Prohormone processing enzymes play an essential role in the secretion of mature and fully functional insulin. Defects in insulin processing enzymes including prohormone convertases 1/3 and 2, and carboxypeptidase E (CPE) can lead to beta-cell stress and hyperproinsulinemia, both of which are features of type 2 diabetes. Despite their importance, the regulation and role of this family of enzymes remain to be fully elucidated. Previously, we demonstrated that lipotoxicity led to the degradation of CPE, but did not affect its related enzyme, carboxypeptidase D (CPD). In this study, we found that CPD was significantly up-regulated by elevated glucose, while CPE was not. Low doses of insulin also increased CPD protein levels, consistent with a role for autocrine signaling. Glucose and insulin did not affect CPD or CPE expression in an alpha-cell line. Furthermore, insulin treatment altered the CPD sub-cellular localization, which was distinct from CPE. Somewhat surprisingly, the loss of CPE did not affect the levels of CPD. Knockdown of CPD exerted no effect on CPE protein levels. In addition, while our previous study demonstrated that even modest reduction of CPE was sufficient to induce beta-cell apoptosis, CPD knockdown did not affect cell viability. Taken together, our data demonstrate that CPE and CPD are differentially localized, differentially regulated and unlikely to have compensatory functions in pancreatic beta-cells. PMID- 21629001 TI - Participants in the workshop. PMID- 21629002 TI - Age at Onset or Late-Life Depression: A Research Agenda Report of a MacArthur Foundation-NIMH Workshop. AB - A workshop titled "Age of Onset in Late-Life Depression" was cosponsored by the MacArthur Foundation Research Network on the Psychobiology of Depression and NIMH in November 1992. The purpose was to better define the relevant methodological and conceptual issues regarding age-at-onset research and to address the ways in which age at onset might more fruitfully inform progress in understanding and treating late-life depression. This paper summarizes the workshop and includes consensus recommendations for future investigation. PMID- 21629003 TI - Negative symptoms in late-life schizophrenia. AB - The authors assessed the relationship of negative symptoms to demographic, other clinical, and neuropsychological variables to investigate and, to an extent, validate the assessment of individual negative symptoms in older schizophrenic patients. Sixty-four subjects meeting DSM-III-R criteria for schizophrenia, including 39 with onset before age 45 (early-onset schizophrenia, or EOS) and 25 diagnosed with late-onset schizophrenia (LOS) were recruited, along with 35 normal comparison (NC) subjects. All the subjects were assessed, using Scales for the Assessment of Negative Symptoms, other psychiatric rating scales, and a comprehensive neuropsychological evaluation. The prevalence of alogia and anhedonia/asociality was comparable in EOS and LOS groups. The prevalence of affective blunting and avolition/apathy was greater in EOS than in LOS. The authors propose a three-factor model of negative symptoms in older, ambulatory, neuroleptic-treated schizophrenic patients. Limitations of the study, as well as possible implications of findings are discussed. PMID- 21629004 TI - MRI volumetric studies in Alzheimer's disease: relationship to clinical and neuropsychological variables. AB - The authors studied 34 subjects with probable dementia of the Alzheimer type (DAT) and 28 healthy, age-matched control subjects, using a 1.5-tesla GE MRI scanner. Absolute volumes of ventricular and sulcal cerebrospinal fluid spaces and volumes, corrected for total intracranial volume, were significantly higher (P < 0.01) in the DAT group, compared to the control group. Right-left measures of hemispheric structural asymmetry were greater in the DAT group, whereas aging alone influenced brain structure in the control group. Also, in the DAT group, certain volumetric measures were significantly correlated with specific clinical indicators of severity of illness and some neuropsychological indices. These data demonstrate widespread structural abnormalities in relatively early DAT and a relationship between brain structure and some measures of cognitive dysfunction. PMID- 21629005 TI - Caregiving for Hispanic Alzheimer's disease patients: understanding the problem. AB - The authors analyze aspects of caregiving for Hispanic Alzheimer's disease (AD) patients. A review of the literature on caregiving and ethnicity and data concerning caregiving for AD patients in different Hispanic groups are presented. The presence of ethnicity-related differences in caregiving for Hispanic AD patients is confirmed. These differences, however, appear to be present largely in certain areas, such as household composition, distribution of caregiving roles, help-seeking behavior, and site of caregiving. The authors consider areas that need to be addressed in future research and specific modifications to current caregiving models for the study of AD patients in the Hispanic community. PMID- 21629006 TI - Sleep Electroencephalographic Studies After ECT. AB - Forty-one patients with major depressive disorder were treated with electroconvulsive therapy (ECT). Sleep polysomnography studies (SPSs) were performed after the course of ECT. The hypotheses tested were that age is a significant factor in post-ECT SPS results and that some SPS parameters are correlates of outcome of ECT. An interaction between age and response to ECT could not be identified; however, older patients demon strated significantly disrupted sleep post-ECT. Response to ECT was associated with lower REM activity and lower REM density. Sleep-onset REM periods post-ECT were observed in almost 50% of the patients regardless of age. The SPS monitoring of recovery after a course of ECT may identify sleep correlates of response to ECT and variables associated with poorer longitudinal outcome. PMID- 21629007 TI - A user-friendly instrument for rating agitation in dementia patients. AB - There are many rating instruments designed to assess symptoms of agitation in patients with dementia. Most scales require that raters assess multiple specific behaviors over several days, limiting the use of such scales to clinical staff who have direct contact with patients but do not have the time to complete a comprehensive assessment following each period of observation. Also, scales that assess behavior over several days must rely on "secondhand" information describing behaviors not directly observed by the rater. The Pittsburgh Agitation Scale (PAS) is an easy-to-use instrument, based on direct observations of the patient, that was developed to monitor the severity of agitation associated with dementia. Interrater reliability and measures of validity have been established with clinical staff on a busy psychogeriatric inpatient unit and with research personnel in a nursing home setting. PMID- 21629008 TI - A new scale for comprehensive assessment of psychopathology in Alzheimer's disease. AB - The Dementia Signs and Symptoms (DSS) Scale documents non-cognitive signs and symptoms (e.g., delusions, hallucinations, anxiety, depression, mania, and behavioral disturbances) in dementia. Patients, informants, and a clinical examiner rated signs and symptoms over the preceding month. Fifty-six Alzheimer's disease patients were administered the DSS, the BEHAVE-AD, the Cornell Scale for Depression in Dementia, the Young Mania Rating Scales, the Hamilton Depression Scale, the Hamilton Anxiety Scale, and the Psychogeriatric Dependency Rating Scale. DSS subscale scores correlated with corresponding scale scores, confirming construct validity. The DSS subscales were internally consistent (Cronbach's alpha, 0.3 7-0.75) and interrater reliability was high (ICC, 0.92-0.99). PMID- 21629009 TI - Childhood abuse history in older women with panic disorder. AB - Seventeen female subjects (ages 55-73) participating in a clinical trial for older panic disorder patients were administered the Childhood Trauma Questionnaire. Seven subjects reported histories of sexual abuse. Physical abuse was reported by 10 subjects, including six of the seven who also reported sexual abuse. The data raise questions regarding the possible contributory role of childhood trauma in some panic disorder patients. PMID- 21629010 TI - A naturalistic study of trazodone in the treatment of behavioral complications of dementia. AB - Altered serotonin transmission has been associated with behavioral complications of degenerative dementia. This study examined whether trazodone, a serotonergic antidepressant, may be useful in treating behavioral syndromes associated with dementia. Twenty-two dementia patients with behavioral problems were treated with trazodone (mean daily dose 172 +/- 107 mg; mean duration 20 days). Seventy of cognitive impairment and behavioral symptoms were rated using the Mini-Mental State Examination (MMSE), the Global Assessment Scale, the Hamilton Rating Scale for Depression, and the Brief Psychiatric Rating Scale. A global assessment of improvement was made by chart review. Mean scores, with the exception of the MMSE, improved modestly but significantly between admission and discharge. Chart review of target symptoms revealed that 82% of the patients showed moderate-to marked improvement on discharge, with most able to return to their preadmission residence. Trazodone was generally well tolerated except for occasional mild sedation effects. Results suggest that trazodone may be a useful alternative to neuroleptics or benzodiazepines in treating behavior problems in some demented patients. PMID- 21629011 TI - On early Alzheimer's disease presenting as pseudoseizures. PMID- 21629012 TI - Stigma in America: has anything changed? Impact of perceptions of mental illness and dangerousness on the desire for social distance: 1996 and 2006. AB - Data from the 1996 and 2006 General Social Survey were analyzed to examine the relationship between the desire for social distance from individuals with mental illness and a number of factors that were thought to contribute to it, including perceptions of mental illness and dangerousness. Random samples of participants were assigned to one of four experimental conditions, in which they were read a vignette describing a character who presented with alcoholism, depression, schizophrenia, or minor problems. The desire for social distance from characters whose presenting problems were alcoholism or depression was significantly lower in 2006 than in 1996. The participants' perceptions that the character was mentally ill and/or dangerous to others partially mediated the association between presenting problem and social distance. Participants who were younger, white, better educated, and attended religious services more often required less social distance from the vignette characters than did their counterparts. PMID- 21629013 TI - Cognitive functioning in recent onset psychosis. AB - The purpose of the study was to compare the cognitive functioning of persons with a recent onset of psychosis with schizophrenia-spectrum disorders and bipolar disorder and nonpsychiatric controls. A total of 56 persons with a schizophrenia spectrum disorder and 60 with bipolar disorder, all with a recent onset psychosis, and 312 nonpsychiatric controls were evaluated using the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS) and the Wisconsin Card Sorting Test (WCST). Comparison of the three groups through analysis of covariance indicated a significant difference among the groups for all of the cognitive variables. Pairwise contrasts of the two recent onset groups showed a significant difference favoring the bipolar disorder group on RBANS Language (p = 0.020) and Total (p = 0.050) and a marginally significant difference on RBANS Immediate Memory (p = 0.053) but not on the other RBANS variables or on the WCST. Cognitive performance is broadly impaired in recent onset psychosis, with a cognitive advantage to bipolar disorder patients compared with schizophrenia spectrum patients. PMID- 21629014 TI - A controlled comparison of the effectiveness and efficiency of two psychological therapies for posttraumatic stress disorder: eye movement desensitization and reprocessing vs. emotional freedom techniques. AB - The present study reports on the first ever controlled comparison between eye movement desensitization and reprocessing (EMDR) and emotional freedom techniques (EFT) for posttraumatic stress disorder. A total of 46 participants were randomized to either EMDR (n = 23) or EFT (n = 23). The participants were assessed at baseline and then reassessed after an 8-week waiting period. Two further blind assessments were conducted at posttreatment and 3-months follow-up. Overall, the results indicated that both interventions produced significant therapeutic gains at posttreatment and follow-up in an equal number of sessions. Similar treatment effect sizes were observed in both treatment groups. Regarding clinical significant changes, a slightly higher proportion of patients in the EMDR group produced substantial clinical changes compared with the EFT group. Given the speculative nature of the theoretical basis of EFT, a dismantling study on the active ingredients of EFT should be subject to future research. PMID- 21629015 TI - Prevalence, comorbidity, and correlates of DSM-IV axis I mental disorders among female university students. AB - This cross-sectional study evaluated the prevalence of Diagnostic and Statistical Manual of Mental Disorders, 4th edition (DSM-IV), axis I mental disorders among Spanish female students and investigated their psychiatric comorbidity and correlates. 1054 female students with a mean age of 22.2 years were randomly selected, with stratification by academic seniority and the type of academic discipline. The cases of mental disorder were identified by clinically trained interviewers with the aid of the Structured Clinical Interview for DSM-IV Axis I Disorders-Clinician Version. The lifetime prevalence of the targeted psychiatric disorders was 50.8%, and its point prevalence was 37.3%. The commonest disorders were nicotine dependence, depression, and generalized anxiety disorder. Nearly 37% of subjects with a psychiatric disorder had two or more diagnoses. Mental illness was associated with family income, financial independence, type of academic discipline, violence from men, social support, and self-esteem. Psychiatric disorders are common among female university students. Serious attention should be paid to preventive and therapeutic programs in this group. PMID- 21629016 TI - Interaction between alliance and technique in predicting patient outcome during psychodynamic psychotherapy. AB - The current study examined whether alliance interacted with psychodynamic interventions to predict patients' psychotherapy outcomes. A prospective study of psychodynamic psychotherapy with 68 outpatients who were treated by 23 therapists was used. The patients rated the alliance with their therapist early in treatment. Therapist use of psychodynamic techniques was reliably rated by independent clinicians for the same sessions. The therapy outcomes were measured at the end of treatment based on the patients' global symptomatology as well as estimate of improvement across a broad range of functioning. In all models, we controlled for the patients' pretherapy psychiatric severity. Analyses were conducted using multilevel modeling to account for therapist effects. Results revealed that patient rated alliance was significantly related to improvement on a measure of broad band functioning. In addition, alliance and psychodynamic interventions interacted to predict this scale of multidimensional therapy outcome. Further, results showed that several individual psychodynamic techniques interacted with alliance that were meaningfully related to this measure of broad band outcome including (1) linking current feelings or perceptions to the past; (2) focusing attention on similarities among patient's relationships repeated over time, settings, or people; and (3) identifying recurrent patterns in patient's actions, feelings, and experiences. In this sample of outpatient psychodynamic treatments, the dynamic techniques were most effective when provided in the context of strong alliances. PMID- 21629017 TI - Comparison of anxiety-related traits between generalized and nongeneralized subtypes of social anxiety disorder. AB - This study aimed to investigate the possible difference in anxiety-related traits between the generalized and nongeneralized subtypes of social anxiety disorder (SAD). Two hundred seventy-three SAD Korean outpatients completed the Anxiety Sensitivity Index (ASI), the Trait Form of the State-Trait Anxiety Inventory (STAI-T), Retrospective Self-Report of Inhibition (RSRI), and the Liebowitz Social Anxiety Scale (LSAS) as part of their assessments. The unadjusted total scores of the ASI, STAI-T, RSRI, and LSAS differed between the two subtypes, according to an independent t-test. However, this result was not significant (ASI: F = 2.363, p = 0.127; STAI-T: F = 0.004, p = 0.949; RSRI: F = 1.518, p = 0.220) after adjusting for LSAS total score. The comparison of anxiety-related traits did not show any difference between the subtypes after adjusting for illness severity. These results may suggest that the two SAD subtypes are on a continuum of the same illness, differentiated only by symptom severity. PMID- 21629018 TI - Measurement of well-being in the workplace: the development of the work well being questionnaire. AB - Because there has been a lack of a single comprehensive measure for assessing workplace well-being, we elected to develop such a self-report measure. Provisional items were extracted from the literature on "positive psychology" and were adapted to capture their workplace application. The provisional 50-item set was completed by a nonclinical sample of 150 adults. A second and third sample was recruited to examine its reliability and any impact of depressed mood and sociodemographic and work-related variables, respectively. Factor analysis identified four domains, "Work Satisfaction," "Organizational Respect for the Employee," "Employer Care," and a negative construct-"Intrusion of Work into Private Life." High test-retest reliability was demonstrated for the final 31 item measure, whereas there was no distinct impact of depressed mood on the scale scores. Work Satisfaction scale scores were influenced by job type. Gender effects were found for two of the four scales, whereas a longer period of employment inversely linked to Organizational Respect for the Employee and Employer Care scores and was conversely associated with higher Intrusion of Work into Private Life scores. The refined measure should enable individuals and employers to quantify the levels of support and well-being provided by employing organizations. PMID- 21629019 TI - Coping with schizophrenia in forensic services: the needs of relatives. AB - This study aimed to assess the needs of the relatives of patients in forensic services and to understand their concerns, perceptions, and ability to cope with the symptoms of schizophrenia. The needs of 18 relatives were assessed using the Family Questionnaire (FQ; Quinn et al., Acta Psychiatr Scand 108:290-296, 2003) and the Relatives' Cardinal Needs Schedule (RCNS; Barrowclough et al., Psychol Med 28:531-542, 1998). The FQ revealed that all symptoms were rated as "frequent." The RCNS supported the data from the FQ because the relatives displayed a need for support and information about the mental illness of schizophrenia. Antisocial behavior was rated as the highest cardinal need (83%), with negative symptoms, interpersonal behavior, psychotic symptoms, and affective symptoms also rated as cardinal needs by more than half of the relatives. The relatives have to cope with a range of symptoms and behaviors displayed by the forensic patient, which can be extremely difficult. Forensic services should provide support for families making information about antisocial behavior a priority. PMID- 21629020 TI - An experience-sampling study of depressive symptoms and their social context. AB - Both clinical and subclinical depression are associated with social impairment; however, few studies have examined the impact of social contact in the daily lives of people with depressive symptoms. The current study used the experience sampling methodology to examine associations between depressive symptoms, social contact, and daily life impairment in 197 young adults. Depressive symptoms were associated with increased isolation, negative affect, anhedonia, and physical symptoms, decreased positive affect, and social and cognitive impairment in daily life. For people with more depressive symptoms, being with social partners who were perceived as close was associated with greater decreases in negative affect, as well as increases in positive affect. Ironically, participants with depressive symptoms reported spending less time with people whom they perceived as close, minimizing the protective effects of socializing. These results suggest that people experiencing depressive symptoms may be especially sensitive to the nature of social interactions. PMID- 21629021 TI - Suicide means used by Chinese rural youths: a comparison between those with and without mental disorders. AB - Reports on Chinese rural youth suicide indicated patterns different from those of the West. Only about 30% to 70% young victims had had diagnoses of psychiatric illnesses (Phillips et al., Lancet 359:835-840, 2002; Xiao et al., Chin J Psychiatry 36:129-131, 2003), and more than 60% of them used pesticides as suicide means (Wang et al., Lancet 372:1765-1773, 2008). To prevent suicides in rural China, it is important to know the choice of means by Chinese young suicide victims with and without mental disorders. Data on suicide cases in China's rural areas gathered from a big psychological autopsy study were studied for demographic characteristics, suicide methods, and the presence of mental disorders. The findings in the suicide victims with and without mental disorders showed significant differences in suicide method selecting. Victims with mental disorders tended to select violent methods compared with those without mental disorders (31.4% vs. 16.2%). Hanging is method more likely chosen by the mentally disordered victims (13.3%) than those without a mental disorder (7.8%). Mental status affects the means choice among the Chinese rural young suicide victims. Among them, the female victims without mental disorders tended to act on impulsivity and used nonviolent means such as pesticide consumption for suicide. This study informs suicide prevention measures in both China and rest of the world. PMID- 21629022 TI - Transient global amnesia as a revealing sign of giant transtentorial meningioma: case report and review of the literature. AB - Since 1974, only 14 papers in MEDLINE-indexed journals have linked transient global amnesia (TGA) to a brain tumor. Among these, two described the entity of transient epileptic amnesia (TEA), whereas most reports lacked the functional investigations that might differentiate the two conditions. Only six cases have linked TGA or TEA to a meningioma, and none of them were transtentorial. We report the case of a 75-year-old woman who presented with one episode of TGA as a sole symptom of a giant transtentorial meningioma in the right hemisphere. A sudden attack of TGA with a rather typical clinical presentation--anterograde amnesia with selective retrograde features that lasted for a few hours, with stereotypic questions, no associated symptoms or neurological impairment, and no recurrence--occurred while she was at home; this was witnessed by her husband. The neurological examination was unremarkable, with the exception of a slight left homonymous superior quadrantonopia. Brain imaging (computed tomography and magnetic resonance imaging) showed a huge meningioma originating from the right tentorium, extending from the cerebellar hemisphere to the midst of the temporal lobe. An electroencephalogram did not reveal temporal spikes. The tumor was completely excised, and she has remained asymptomatic for 6 months. Although TGA is generally a functional entity, without an organic substrate to prompt surgical sanction, a full investigation may be warranted to rule out the possibility of a silent intracranial tumor. PMID- 21629023 TI - The impact of detecting bipolar disorder in previously diagnosed unipolar patients at a specialist depression clinic. AB - This study aimed to examine the short-term clinical impact of identifying bipolar disorder in patients previously managed as having a unipolar disorder. The study was incorporated within a consecutive sample of 1000 patients attending a specialist depression clinic for diagnostic and management considerations. Of those assessed, 34% were evaluated as having a bipolar disorder, with this condition having been diagnosed for the first time in three-quarters of those patients. We reviewed sample members 12 weeks later and compared the courses of the "newly diagnosed" and "established" bipolar subsets. Some four-fifths of the bipolar patients reported a degree of improvement, whereas there were no clear differences between the two bipolar subsets. The nondifferential outcome of the bipolar (previously and newly diagnosed) subsets could suggest that there were nonspecific benefits of assessment or that the management was optimized for both groups. Future studies examining the impact of diagnosing a bipolar disorder would therefore benefit from the close consideration of the optimal control group or control strategy. PMID- 21629024 TI - Coping styles in carers of people with recent and long-term psychosis. AB - Avoidant coping is associated with distress in carers of people with psychosis. We hypothesized that this form of coping would abate as carers adapt their coping strategies in the course of the illness. One hundred and forty-one carers of two groups of patients, with recent onset and longer established psychosis, respectively, completed self-report measures of coping and general distress. We found that avoidant coping strategies were associated with carer distress but not with duration of illness. These results argue the need for interventions to reduce the carers' reliance on maladaptive (avoidant) strategies at any stage of the illness. PMID- 21629025 TI - Mediumship research. PMID- 21629027 TI - Getting "research rich" at a community hospital. PMID- 21629028 TI - Understanding the measure applications partnership. PMID- 21629029 TI - Is your facility struggling with millennial nurse retention? PMID- 21629031 TI - Future of nursing special: Practicing to potential. PMID- 21629032 TI - When time isn't on your side: 12-hour shifts. PMID- 21629033 TI - The role of the corporate chief nursing officer. PMID- 21629034 TI - Outpatient nurses "own" MRSA prevention initiatives. PMID- 21629035 TI - Using a blog to improve communication. PMID- 21629036 TI - Coping with downsizing; deciding to pursue Magnet(r) designation. PMID- 21629037 TI - Lipid targets during statin treatment in dyslipidemic patients affected by nonalcoholic fatty liver disease. AB - INTRODUCTION: Nonalcoholic fatty liver disease (NAFLD) is associated with both dyslipidemia and increased risk for cardiovascular disease. Despite the indication to treat in patients affected by both dyslipidemia and NAFLD, an undertreatment in statin therapy due to the potential liver damage is frequently observed. We sought to evaluate retrospectively the impact of statin on the lipid profile and on the achievement of low-density lipoprotein (LDL) cholesterol targets in relation to the National Cholesterol Education Program--Adult Treatment Panel III-cardiovascular risk in dyslipidemic patients presenting with a clinical--diagnosis of NAFLD and elevated liver enzymes before statin prescription. As a secondary endpoint, the authors investigated whether statin could be associated with changes of liver enzymes. METHODS: Forty-three patients with dyslipidemic NAFLD presenting with increased values of aspartate aminotransferase and/or alanine aminotransferase and/or gamma-glutamyl transferase at baseline were analyzed retrospectively as regard the lipid profile and liver enzymes (values reported before statin and during statin therapy). RESULTS: Total cholesterol, LDL and triglycerides were significantly reduced at follow-up (5.4 +/- 5.4 months). The LDL target was achieved at the second visit in 30 patients (69.8%).The number of patients achieving the LDL target was significantly higher in low-risk group compared with moderate- and high-risk subjects. Liver enzyme levels showed no significant changes between baseline and follow-up. CONCLUSIONS: Statin treatment was effective (without changes in liver enzymes) in patients with dyslipidemia and NAFLD and therefore, affected by a profound alteration in lipoprotein metabolism. The number of patients achieving LDL target was related to the Adult Treatment Panel III risk classification, being higher in patients with lower risk. PMID- 21629038 TI - Enhancer of zeste homolog 2 expression is associated with tumor cell proliferation and invasion in cervical cancer. AB - INTRODUCTION: Enhancer of zeste homolog 2 (EZH2) has been reported to be associated with biological malignancy in several cancers. High expression of EZH2 is associated with tumor cell proliferation, invasion and metastasis and has important clinicopathologic significance. However, little is known about the function and mechanisms of EZH2 in cervical cancer. METHODS: This study was conducted to investigate the clinical value of EZH2 expression in cervical cancer and the association between EZH2 and possible downstream proteins using EZH2 targeted interfering RNA. RESULTS: EZH2 expression rate was significantly greater in cervical cancer tissue than in normal cervical tissue both in mRNA and protein level. High EZH2 expression was significantly associated with differentiation, depth and lymphatic invasion. EZH2 gene RNAi resulted in inhibition of cell growth, cell cycle arrest and decreased ability of invasion in C33A cells. PCNA and MMP-2 expressions were downregulated, whereas p21 was upregulated after inhibition of EZH2. CONCLUSIONS: High EZH2 expression may be associated with tumor cell proliferation and invasion in cervical cancer, and that EZH2 may be one of the candidates for new molecular therapeutic targets for the treatment of cervical cancer. PMID- 21629039 TI - Differences in drug-eluting stents used in coronary artery disease. AB - The introduction of drug-eluting stents (DES) has improved the efficacy of percutaneous coronary intervention by addressing the issue of neointimal proliferation, a pathology contributing to restenosis. First-generation stents eluting sirolimus or paclitaxel were joined by second-generation stents, such as the everolimus- and the zotarolimus-eluting stents, promising increased safety and efficacy. As a result, there is a plethora of drug-eluting stents available, with differences in the stent platform, the polymer coating and the eluted drug, which translate into differences in biological markers of efficacy, such as late loss. However, it remains controversial whether these discrepancies have an impact on clinical markers of safety and efficacy, or if the improved efficacy of DES is a class effect. This article reviews the differences between DES by looking into the biological differences and into trials and registries of DES. PMID- 21629040 TI - Predictive value of body mass index at age 18 on adulthood obesity: results of a prospective survey of an urban population. AB - INTRODUCTION: Overweight children often become obese adults. The main purpose of this study was to define the risk of teenage obesity as a predictor of adulthood obesity in an under-studied and urban population. A secondary aim was to determine whether gender had an effect on the predictive value of teenage obesity. The final goal was to examine the influence of dietary habits on obesity. METHODS: This cross-sectional survey was conducted using convenience sampling of an urban, primarily African American, community. Demographic questions were supplemented by a dietary history and measurement of body anthropometrics. RESULTS: Five hundred three subjects were interviewed; 86% were African American, and the mean age was 43 years. Body mass index (BMI) at age 18 was the strongest factor associated with current obesity status in univariate and multivariate analyses. Sixty-nine (13.7%) subjects were overweight at age 18, and 28 (5.6%) were obese. Of the 28 obese teenagers, 22 (78.6%) went on to become obese as adults. Only 2.1% of nonobese adults were obese as teenagers. For women, BMI at age 18 was more predictive of adult BMI than for men. Gain of >= 5 BMI units after age 18 was linked to a higher prevalence of diabetes and hypertension. No dietary differences were found between obese and nonobese adults. CONCLUSIONS: The results of this study provide evidence that high BMI at age 18 is strongly correlated with adulthood obesity, much more so with women than men. Eating habits did not have an impact, suggesting that obesity may be the result of a combination of factors yet to be clearly defined. PMID- 21629042 TI - Systemic mastocytosis. AB - Systemic mastocytosis (SM) is a clonal disorder of hematopoietic system characterized by abnormal growth and accumulation of mast cells in various tissues. Its clinical spectrum ranges from mild disease to an aggressive course with life-threatening conditions. Some of the clinical signs or symptoms of SM (hyperhidrosis, syncope and hypotensive/tachycardiac attacks) require consideration of pheochromocytoma and carcinoid syndrome in the differential diagnosis. The diagnosis relies on the demonstration of mast cell aggregates in bone marrow or extracutaneous tissues. The World Health Organization categorizes SM into 6 variants: indolent SM, SM with associated clonal hematological nonmast cell lineage disease, aggressive SM, mast cell leukemia, mast cell sarcoma and extracutaneous mastocytosis. Patients with indolent SM have a favorable prognosis with a life expectancy comparable with the healthy population, and symptomatic treatment is usually sufficient. However, more aggressive forms may be life threatening, and cytoreductive treatment is indicated in most cases. PMID- 21629041 TI - Variants and haplotypes in angiotensinogen gene are associated with plasmatic angiotensinogen level in Mexican population. AB - INTRODUCTION: The plasmatic angiotensinogen (AGT) level has been associated with essential hypertension. Linkage analysis has found a relationship between the AGT gene locus and hypertension in the Mexican-American population, but studies have failed to identify genetic variants associated with hypertension or plasma AGT levels. This study analyzes the relationship between polymorphisms in the AGT gene and plasmatic AGT levels in Mexican population. METHODS: Nine polymorphisms in AGT gene were genotyped, and plasma AGT level was determined by enzyme-linked immunosorbent assay. RESULTS: Differences in AGT plasma levels were associated with 2 polymorphisms: T-20G, TT = 25.3 +/- 8.3 versus TG + GG = 21.6 +/- 8.8 MUg/mL; P = 0.008 and C3389T (T174M), CC = 25.8 +/- 9.9 versus TC + TT = 20.5 +/- 5.4 MUg/mL; P = 0.0002. Haplotype 2 was associated with low plasma AGT (-5.1 MUg/mL [95% confidence interval: -8.6 to -1.6], P = 0.004) and Haplotype 8 was associated with high plasma AGT (6.5 MUg/mL [95% confidence interval: 2.5 to 10.6], P = 0.001). This association remained after adjustment for covariates. A Likelihood Ratio Test for haplotype-phenotype association adjusted for covariates resulted in chi = 38.9, P = 0.0005. The total effect of the haplotypes on plasma AGT level variance was 19.5%. No association was identified between haplotypes and quantitative traits of blood pressure. CONCLUSIONS: Two polymorphisms (T-20G and C3389T) and 2 haplotypes (H2 and H8) showed an association with plasma AGT levels in Mexican population. PMID- 21629043 TI - Evaluation and management of hip fracture risk in the aged. PMID- 21629044 TI - Depression, screening and quality of life in chronic kidney disease. AB - INTRODUCTION: To determine the prevalence of positive screens for depression and to assess quality of life (QoL) and usefulness of the brief and easily administered Patient Health Questionnaire-2 (PHQ-2) for depression screening in the chronic kidney disease (CKD) population; and to explore the relationship between depressive symptoms and markers of inflammation. METHODS: Seventy-one adult patients with estimated glomerular filtration rate <60 mL/min/1.73 m or proteinuria, but not on dialysis, were enrolled. QoL was assessed using the Short Form-36. The Center for Epidemiological Studies Depression Scale (CES-D) and PHQ 2 were used to screen for depression. Serum ferritin, albumin, C-reactive protein and hematocrit were also measured as markers of inflammation. RESULTS: The PHQ-2 and CES-D were significantly correlated (P < 0.05). Positive scores on the CES-D or PHQ-2 had significantly lower Short Form-36 scores. Mean hemoglobin values were significantly lower in patients who screened positive for depression either by CES-D (12.2 +/- 1.7 versus 13.2 +/- 1.7, P < 0.05) or by PHQ-2 (12 +/- 1.6 versus 13.4 +/- 1.6, P < 0.01). Neither PHQ-2 nor CES-D correlated with other markers of inflammation in this sample. CONCLUSION: Both the CES-D and the PHQ-2 can identify patients with CKD who need further evaluation for depression. The PHQ-2 seems to be a useful screen for depression and impaired QoL in a renal clinic setting. Patients with CKD and lower hemoglobin may be at greater risk for depression than those with normal values. PMID- 21629045 TI - Peripheral nerve defect repair with epineural tubes supported with bone marrow stromal cells: a preliminary report. AB - INTRODUCTION: This study was performed to assess the effects of nerve gap repair with isogenic epineural tubes filled with isogenic bone marrow stromal cells (BMSC) as an alternative to autograft repair. METHODS: A total of 24 epineural tubes were transplanted as a conduit to bridge 20 mm nerve gap defects in 2 experimental groups. In group 1, the tube was filled with saline, whereas in group 2 with isogenic BMSC prestained with PKH-dye. In all, 12 autograft sciatic nerve repairs served as a control. Sensory and motor recovery was evaluated by gastrocnemius muscle index; pinprick, toe-spread tests; and somatosensory-evoked potentials at 6 and 12 weeks postrepair. Histomorphometry and immunostaining were also performed. RESULTS: Evidence of successful nerve regeneration was present in all animals at 6 and 12 weeks. There were no significant differences between experimental groups in functional recovery--toe-spread and pinprick tests; however, epineural conduit groups trended toward better toe-spread scores compared with autograft repair at 6 weeks. All animals had full sensory recovery as evaluated by pinprick at 12 weeks. Saline group had significantly higher gastrocnemius muscle index compared with other groups at 6 weeks; however, no differences were noted at 12 weeks. Histomorphometrical assessment did not reveal superiority of any group at 6 weeks postrepair. However, at 12 weeks, the BMSC group had significantly increased myelin thickness compared with other groups and larger diameter nerve fibers than autograft. In group 2, PKH-positive cells and expression of nerve growth factor, Laminin B2, glial fibrillary acidic protein, and vascular endothelial growth factor were confirmed. nerve growth factor- and glial fibrillary acidic protein-positive BMSC were also found inside the tube. CONCLUSION: The epineural tube is a viable, naturally occurring biologic conduit for nerve repair. Cotransplantation of BMSC-enhanced nerve regeneration by means of increased myelinization and expression of neurotrophic factors. Overall, obtained results with epineural tube/BMSC construct were comparable to autograft repair. PMID- 21629046 TI - An update on wound healing and the nervous system. AB - Impaired wound healing is a significant clinical, economic, and social problem. There is a growing body of research over the past several decades that supports the importance of neuropeptides and neurotransmitters in wound healing, particularly in diabetic patients. The purpose of this article is to review current translational research that supports the role of the nervous system in normal wound physiology and the current state of clinical application. PMID- 21629047 TI - Delayed autologous breast reconstruction after postmastectomy radiation therapy: is there an optimal time? AB - The optimal time for delayed autologous breast reconstruction after postmastectomy radiation therapy (PMRT) is unknown. Although most reconstructive surgeons recommend waiting for 6 months, this timing is arbitrary. A retrospective analysis was performed of 199 patients undergoing delayed autologous reconstruction; 100 patients had prior PMRT, whereas 99 patients had no previous radiation. Radiated patients had higher overall complications (40% vs. 20.2%, P = 0.0023), including wound dehiscence (11% vs. 3%, P = 0.049), and trended toward increased postsurgical infections (7% vs. 1%, P = 0.065). Logistic regression models of unequally distributed variables found radiation therapy to be the only independent risk factor for wound dehiscence (odds ratio, 3.97; P = 0.04). Mean follow-up for radiated and nonradiated patients was 33.3 months and 39.4 months, respectively. After PMRT, 17 patients were reconstructed within 6 months and 83 after 6 months. No significant differences in complications were observed between these groups. An alternate analysis examined 51 patients reconstructed within 12 months of PMRT and 49 patients reconstructed after; again, there were no differences in complications. As overall complications are similar in patients reconstructed early or late after PMRT, autologous breast reconstruction can potentially be performed earlier than is the current accepted practice. PMID- 21629048 TI - Complications following reconstruction of soft-tissue sarcoma: importance of early participation of the plastic surgeon. AB - BACKGROUND: Soft-tissue sarcomas (STSs) are rare malignant mesenchyme-derived tumors arising most frequently in the extremities. Current treatment involves wide excision and radiotherapy. Reconstruction of defects following limb-sparing surgery is best achieved with free flaps immediately after primary resection. Nevertheless, high rates of wound complications are expected, mainly due to postoperative radiotherapy. Patients inadequately treated with multiple surgeries and repetitive radiotherapy, are more prone to develop complications leading to worst functional outcomes. METHODS: A retrospective analysis of patients referred for reconstruction following STS resection was performed. Patients were classified in the following 2 groups: group A, immediate reconstruction within 1 year since oncologic surgery and group B, delayed reconstruction after 1 year. Statistical analysis was carried out using Student t test and chi2. Odds ratio with 95% confidence interval was estimated. RESULTS: A total of 30 patients were referred to our unit for reconstruction, with 14 patients corresponding to group A and 16 to group B. Significant difference was observed in complication rates between the groups (P < 0.05). Early reconstruction decreased the risk of complications (odds ratio, 0.06; 95% confidence interval, 0.01-0.36). CONCLUSIONS: STSs are best treated in specialist centers. Early referral is highly recommended to achieve good oncologic outcomes. Our results show a clear tendency of higher wound complication rates in patients lately referred for reconstruction. We believe that early involvement of plastic surgeons in the management of STS is mandatory. PMID- 21629049 TI - A closed phalangeal neck fracture with 180-degree rotation of the phalangeal head and concurrent displaced Salter-Harris II fracture. AB - Closed type III phalangeal neck fractures with 180-degree rotation is a rare iatrogenic injury that occurs following failed attempts at closed reduction. Prior to closed reduction, the phalangeal head is in 90-degree rotation. Longitudinal traction during closed reduction then converts the deformity into 180-degree rotation. We present the first documented noniatrogenic case of phalangeal neck fracture with 180-degree rotation that was also associated with a displaced Salter-Harris II fracture at the same joint. The blood supply of the phalangeal head in that scenario is discussed along with precautions that should be taken during open reduction and internal fixation to avoid avascular necrosis of the phalangeal head. PMID- 21629050 TI - Saw injuries causing phalangeal neck fractures in adults. AB - The outcome of adult phalangeal neck fractures caused by saws has not been previously investigated. Over a 15-year period, a total of 36 cases of saw related injuries resulting in phalangeal neck fractures of the middle phalanx of the finger or the proximal phalanx of the thumb were treated. All injuries were industrial and covered by insurance. They were all adult males with type II fractures. The fracture were managed by K-wire fixation, immobilizing the distal joint only. The proximal joints were mobilized immediately after surgery and the K-wire was removed after 5 weeks. The following 2 patterns of saw injuries were observed: dorsal impact injuries resulting in a phalangeal neck fracture and concurrent extensor tendon injury (group I, n = 20 patients), and dorsolateral impact injuries resulting in concurrent extensor (+/- partial flexor) tendon and nerve injury (group II, n = 16 patients). The outcome with respect to range of motion and return to work was significantly better in the former group. Major complications such as nonunion, malunion, avascular necrosis of the phalangeal head, osteomyelitis, or contractures of the proximal interphalangeal joints were not observed. Two patients (1 in each group) had superficial pin-site infection. It was concluded that adult phalangeal neck fractures caused by saws is a separate entity from the previously reported series of closed phalangeal neck fractures in adults. PMID- 21629051 TI - Use of inferior gluteal artery and posterior thigh perforators in management of ischial pressure sores with limited donor sites for flap coverage. AB - BACKGROUND: Reconstructive surgery for ischial pressure sore defects presents a challenge because of high rates of recurrence. The aim of this study was to describe the use of inferior gluteal artery (IGA) and posterior thigh perforators in management of ischial pressure sores with limited donor sites. PATIENTS AND METHOD: Between September 2005 and 2009, 11 patients (9 male, 2 female) with ischial sores were operated by using IGA and posterior thigh perforator flaps. The data of patients included age, sex, cause of paraplegia, flap size, perforator of flap, previous surgeries, recurrences, complications, and postoperative follow-up. RESULTS: Nine IGA and 5 posterior thigh perforator flaps were used. Six patients presented with recurrent lesions, 5 patients were operated for sacral and contralateral ischial pressure sores previously. In 2 patients, IGA and posterior thigh perforator flaps were used in combination. Patients were followed for an average of 34.3 months. In 2 recurrent cases, readvancement of IGA perforator flap and gluteus maximus myocutaneous flap were treatment of choice. CONCLUSION: Treatment of patients with recurrent lesions or multiple pressure sores is challenging because of limited available flap donor sites. In this study, posterior thigh perforator flaps were preferred in patients in whom the previous donor site was the gluteal region. IGA perforator flaps were the treatment of choice in patients for whom posterior thigh region was previously used. Alternately, preserved perforators of previous conventional myocutaneous flaps enabled us to use these perforators in recurrences. PMID- 21629052 TI - Coumadin-induced skin necrosis of the breasts: case report. AB - A case report of coumadin-induced skin necrosis (CISN) is presented, followed by a topic review of CISN, which reviews presentation, pathophysiology, differential diagnosis, prevention, and management of this disorder. The prevalence of CISN is low (0.01%-0.1% of patients receiving coumadin). However, of those affected, over 50% required some form of surgical debridement or reconstruction. Although skin necrosis secondary to coumadin therapy is rare, it is essential for plastic surgeons to be aware of this clinical entity in order to make the correct diagnosis and provide appropriate treatment. PMID- 21629053 TI - Two-staged tendon reconstruction in flexor tendon ruptures secondary to fracture of the hamate hook. AB - We report a case of 2-staged reconstruction of flexor tendons ruptured spontaneously by attrition. A 49-year-old man presented with inability to flex the ring and little fingers of his left hand. Preoperative computed tomographic scans revealed fracture of the hamate hook. At the time of the operation, both the flexor superficialis and profundus of the little finger and the flexor profundus tendon of the ring finger were ruptured adjacent to the fracture site of the hamate. Because the flexor tendon rupture secondary to the fracture of the hamate is extremely rare, and surgical outcomes of previous reports are not satisfactory, a decision was made to perform 2-staged reconstruction of ruptured flexor tendons. The surgical result was excellent with complete restoration of full range of motion. This report describes for the first time to our knowledge, the technique, and rehabilitation of 2-staged tendon reconstruction in a patient with hook of hamate fracture. PMID- 21629054 TI - A new model of acquired lymphedema in the mouse hind limb: a preliminary report. AB - Lymphedema is known to be caused by many pathologic conditions; however, its correct diagnosis and optimal therapeutic strategies remain to be established. In this report, we describe an experimental model for acquired lymphedema in the lower extremity of the mouse that creates a lymphatic block in the groin induced by both radiation treatment and surgical division of the superficial and deep lymphatics. To evaluate the lymphatic system in this model, an indocyanine green fluorescence-sensitive camera system was used. This model has the advantages of relative technical simplicity and cost-effective use of a rodent animal model. Furthermore, a greater range of research tools such as antibodies and various databases are available for mice. This mouse model may be useful to anyone modeling lymphedema mechanisms, by providing a defined molecular context. PMID- 21629055 TI - Fingertip reconstruction with a flap based on the dorsal branch of the digital artery at the middle phalanx: a simple and reliable flap. AB - BACKGROUND: It is of utmost importance for hand surgeons to rehabilitate injured fingertips aesthetically and functionally. The purpose of our study was to investigate the effect of the dorsal branch (DB) finger flap on fingertip reconstruction. METHOD: We used the flap based on the DB of the proper digital artery for fingertip reconstruction in 3 patients at the primary stage. The size of the flaps ranged from 1.5 * 2.0 cm to 2.5 * 4.5 cm. RESULT: The DB finger flaps in our series survived uneventfully during the follow-up period (range, 4-6 months). All the patients were contented with the aesthetic and functional outcomes of the flaps. The average static and moving 2-point discrimination of the flaps was 8.5 mm and 7.2 mm, respectively. CONCLUSION: This sort of flap based on the DB of the proper digital artery was a simple and reliable alternative to reconstruct fingertip defects at the primary stage. PMID- 21629056 TI - Preliminary experiences in treating infantile hemangioma with propranolol. AB - The aim of this study was to evaluate the outcome of treating infantile hemangioma with the use of oral propranolol. A total of 27 patients with hemangiomas were treated with oral propranolol therapy. The subjects included 21 female patients and 6 male patients whose age ranged between 3 weeks and 7 months. Locations of lesions were as follows: 22 on the face and neck, 3 on the trunk, and 2 on the limbs. The dose of 0.5 mg/kg/d of propranolol was administered; and was gradually doubled to a maximum of 2 mg/kg/d. The treatment lasted for a period of 2.75 to 5.75 months without major complications. Two days later, a change in color was observed in 100% of patients, and 2 weeks later >75% reduction in diameter of the original lesion was found in 25.9% of patients. Treating infantile hemangioma with the use of oral propranolol is effective and reliable. PMID- 21629057 TI - Autologous fat grafting for correction of unaesthetic scars. AB - BACKGROUND: Autologous fat grafting has many clinical applications, and its use in Plastic Surgery is increasing. Currently, autologous fat grafts are used in breast surgery, facial rejuvenation, and facial lipoatrophy secondary to antiretroviral therapy and as a treatment for liposuction sequelae, buttock augmentation, and congenital facial hemiatrophy. Their use is expanding rapidly, and their applications in other fields are an ever growing interest within the Plastic Surgery community. OBJECTIVE: To introduce a new application of lipoinjection for the correction of unaesthetic, retracted, or sunken scars. METHODOLOGY: The study consisted of a total of 8 patients (6 women and 2 men), with a mean age of 47 years old, all of whom presented retractile and dystrophic scars in the abdomen (n = 3), arm (n = 1), male breast (n = 1), and face (n = 3). They all received treatment with a fat injection using Coleman technique. General anesthesia was used in 3 patients; deep intravenous sedation plus local anesthesia was used in the remaining 5 patients. A COL-ASP15 cannula was used to harvest the fat and a blunt-tipped COL-19 cannula (Byron Medical) to release the fibrosis and retraction, and for the fat grafting injection. A 4-grade visual scale was use to evaluate the results. RESULTS: An improvement in the scar was achieved in all patients. One operation was required in 5 cases, and 2 operations in 3 cases. There were no complications in any patient and the results were lasting in all cases (the mean follow-up period was of 18 months). CONCLUSIONS: Autologous fat grafting is a good method for correction of retracted or sunken scars instead of the traditional scar surgical excision. PMID- 21629058 TI - Desmoid tumors of the bilateral breasts in a patient without Gardner syndrome: a case report and review of literature. AB - Desmoid tumors constitute 0.02% to 0.03% of all tumors. Consequently, few case reports exist for breast desmoids, even fewer identifying bilateral disease. We present a case report of a patient with bilateral breast desmoids and shoulder desmoid without evidence of FAP or Gardner syndrome. This case report explores the clinical, radiographic, pathologic, and treatment elements for desmoid tumors as well as a review of the literature. PMID- 21629059 TI - The use of prophylactic antibiotics in plastic surgery: update in 2010. AB - BACKGROUND: The indications for prophylactic antibiotics in plastic surgery remain controversial. No recent survey has been reported on the use of prophylactic antibiotics by plastic surgeons in clinical practice. This survey was designed to assess the current use of prophylactic antibiotics by plastic surgeons and to compare trends with previous studies. METHODS: All members of the American Society of Plastic Surgeons with an e-mail address on the Society's website were contacted via an e-mail and sent a link to a SurveyMonkey questionnaire. To survey only in those subspecialty areas that they practice in, surgeons were queried only on the procedures that they perform. Within each section, a list of common representative procedures was included, with questions about the use of antibiotic prophylaxis. RESULTS: A total of 3824 American Society of Plastic Surgeons members were contacted. Of the 3613, 910 with working e-mail addresses responded to the survey for a response rate of 25%. And 833 or 91.5% completed the survey. Survey data cover the percentage of surgeons reporting their use of antibiotics in procedures that they currently perform. The percentage of plastic surgeons who use prophylactic antibiotics in almost all procedures studied has increased significantly when compared with earlier studies. CONCLUSIONS: The use of prophylactic antibiotics by plastic surgeons has increased considerably since the prior studies by Krizek et al (Plast Reconstr Surg. 1975;55:21-32 and 1985;76:953-963). Some of these uses are appropriate because of the use in procedures involving implants and longer operations. The elevated rates for clean procedures are not part of the evidence-based practice. PMID- 21629060 TI - Role of anti-TNF-alpha therapy in fat graft preservation. AB - BACKGROUND/OBJECTIVE: In this study, we evaluated the role of antitumor necrosis factor-alpha (TNF-alpha) therapy in the decrease of adipocyte apoptosis and weight preservation of fat grafts in the rat model. METHODS: A total of 64 rats were randomly divided into 2 groups, with 32 rats in each group. Autologous fat tissue was grafted subcutaneously on the back of each rat. For the experimental group, antirat TNF-alpha monoclonal antibody was injected into the fat grafts during operation. No treatment was given to the tissue in the control group. Eight rats in each group were killed respectively, at days 7, 14, 30, and 60 postoperatively and sampled for assessments of weight preservation, gene expression of TNF-alpha, histology, and adipocyte apoptosis. RESULTS: There were no significant differences in the weight of fat tissues between the control group and the experimental group at days 7, 14, and 30 postoperatively (P > 0.05). However, the preservation ratio of the tissue was 65.36% +/- 14.98% in the antirat TNF-alpha antibody-treated group when compared with the weight at transplantation, which was significantly higher than the control group (44.63% +/ 10.39%) 60 days after the operation (P < 0.05). The numbers of apoptotic cells in the control group were 15.6 +/- 3.17, 24.6 +/- 4.34, 22.8 +/- 2.42, and 27 +/- 3.83 per field at different postoperative intervals. However, the numbers of apoptotic cells in the tissues treated with TNF-alpha antibody were significantly lower than that in the control group, which was 1 +/- 0.63, 4 +/- 1.41, 6 +/- 2.08, and 7.2 +/- 2.82 per field (P < 0.05). Gene expression showed that the expression of TNF-alpha was lower in the experimental group than the control group at days 7 and 14 postoperatively (P < 0.05). CONCLUSION: The results indicate that antirat TNF-alpha monoclonal antibody can preserve the quality of the transplanted fat tissue. PMID- 21629061 TI - A new aspect of metastasis in cutaneous cancer: two cases of presacral squamous cell carcinoma with metastases to internal iliac nodes. AB - We report 2 cases of presacral squamous cell carcinoma. In these cases, it was suspicious that the tumor disseminated to the internal iliac nodes through the direct pathway called the neurovascular lymphatic space (NVLS) around the superior/inferior gluteal vessels or sciatic nerve bypassing external iliac nodes. NVLS was initially reported as accessory pathway which follows the major vessels forming a sheath like structure with an actual or potential space between the vessel wall and the enveloping membrane. NVLS has been reported to be observed as a tubular shadow within the neurovascular sheath by lymphangiographies of lymphedema patients using oil-based contrast material. These cases provide insights into the potential pathway through which a cutaneous tumor disseminates. PMID- 21629062 TI - Interphalangeal and thumb metacarpophalangeal arthrodesis with an intramedullary implant. AB - Arthritis and instability of the finger joints of the hand leads to chronic pain and decreased hand function. In such patients, arthrodesis is a generally accepted surgical treatment. Various techniques are available for arthrodesis of interphalangeal and thumb metacarpophalangeal joints of the hand. We present our experience from 7 patients being treated with interphalangeal and thumb metacarpophalangeal arthrodesis using an intramedullary implant (X-Fuse).Clinical parameters such as finger stability, soft-tissue irritation, infection rate, and nail morphology were recorded. All patients were observed for clinical and radiographic fusion achievement. Complications and hardware failure of the implant was also recorded. The preliminary results from our clinical experience have been positive. Six out of the 7 joints achieved full fusion. The elastic intramedullary implant is an alternative for finger arthrodesis that provides compression, stability, and adequate fusion. Using this technique, our patients regained a pain-free, stable, useful, and cosmetically acceptable hand. PMID- 21629063 TI - Reconstruction of a large tracheal defect using Gore-Tex and external wire stent. AB - Reconstruction of a tracheal defect remains to be one of the biggest challenges in thoracic surgery. We present a case in which we used Gore-Tex and an external wire stent to reconstruct 7 cm of the trachea and obtained satisfactory outcomes. Gore-Tex appears to be a biocompatible, stable, and reversible material that can be safely used to reconstruct tracheal defects. PMID- 21629064 TI - Management of full-thickness abdominal wall defects following tumor resection. AB - OBJECTIVES: Abdominal wall reconstruction continues to evolve with improvement in technique and technology. This study reviews our experience with challenging full thickness (skin, muscle, and fascia) defects following tumor resections often in high-risk patients. METHODS: All patients who underwent abdominal wall reconstruction following full thickness tumor resection were included in the series. Data queried included patient demographics, indications, tumor defect, risk factors, type of repair, complications, and recurrence of the hernia. RESULTS: A total of 30 patients underwent reconstruction after full thickness resection of abdominal wall tumors or tumors of intra-abdominal organs involving abdominal wall. The indications included desmoid tumors (n=6); abdominal wall sarcoma (n=7); colon cancer invading abdominal wall (n=10); pancreatic, ovarian, and retroperitoneal sarcoma with abdominal wall invasion (n=3); and other (n=4). In all, 17 patients underwent simultaneous resection of one or more intra abdominal organs. The type of repair included primary closure (+/-components separation), or mesh-assisted closure (+/-primary fascial closure). Acellular dermal matrix was used more commonly in the patients with tumors of gastrointestinal origin. The average follow-up period was 23 months. Postoperative complications developed in 6 patients (20.6%). Incidence of postoperative complications was higher in patients with colon cancer invading the abdominal wall or those with bowel anastomosis or radiation therapy. An abdominal bulge or hernia developed in 4 patients (13.7%); it was higher in patients who did not have mesh reconstruction. CONCLUSION: Abdominal wall reconstruction after full thickness tumor resection is challenging. It can be performed safely and effectively with attention to surgical technique, patients' risk of infection, and type of mesh. Acellular dermal matrix graft has been a useful addition to minimize morbidity and recurrence in these high-risk patients. PMID- 21629065 TI - Iliac crest flap for mandibular reconstruction after advanced stage mandibular ameloblastoma resection. AB - Ablative surgeries for neoplastic processes of the oral cavity, traumas, infections/inflammations, osteoradionecrosis, and congenital deformities are the most common causes of large mandibular defects. Ameloblastoma is a locally aggressive tumor that, if not treated, can gain an enormous size and cause severe facial disfigurement and functional impairment. Although the smaller lesions of ameloblastoma in the mandible are treated by conservative approaches such as marsupialization, enucleation, and curettage combined with liquid nitrogen spray cryosurgery, larger lesions require radical surgical ablation procedures resulting in large tissue defects. A large mandibular defect has deleterious effects on a person's life, with a significant loss in the quality of life unless it is reconstructed successfully. The aim of present case series report is to show the results of the multidisciplinary treatment of patients with advanced stage ameloblastoma, including tumor resection, simultaneous reconstruction with iliac crest flap, followed by placement of endosseous dental implants, and finally the prosthodontic rehabilitation. PMID- 21629066 TI - Classification of recipient veins in microsurgical flap reconstructions of the lower extremities. AB - It is difficult to cure severe open fractures of the lower extremities due to the frequent occurrence of severe damage to soft tissues or blood vessels, as well as bone defects, and concomitant infection, such as osteomyelitis. If appropriate treatment is not performed, long-term complications, such as nonunion of the bone, intractable ulcers, or other disorders, may arise. Between 1993 and 2010, we reviewed the records of 27 patients, and 28 limbs were treated. A total of 31 free-tissue transfers were performed on these 28 limbs. In this study, we classified the degree of damage based on the operative appearance of the condition of the recipient vein. A state that was normal or near-normal was described as type I-normal. A vein that was buried under a scar, but was comparatively easy to detach and retained a moderate adventitia was classified as type II-moderate. A vein with close adhesion between the wall and the scar that was not easy to detach was described as type III-severe. We also examined the relationships between recipient vein type and each of the following: (1) concomitant injury, (2) bone reconstruction, (3) preoperative infection, (4) length of time from injury to reconstruction, (5) eventual reoperation, and (6) flap necrosis. The results showed that there was a statistically significant relationship between recipient vein type and each of the following: bone reconstruction, preoperative infection, eventual reoperation, and flap necrosis. The selection of recipient vein is a factor that greatly affects the outcome of reconstruction of open fractures of the lower extremities. It is important to preoperatively estimate the condition of recipient vein even though it is difficult to preoperatively assess the condition. In some cases, only recipient veins in poor condition can be found intraoperatively on the affected leg. Under such circumstances, a cross-leg flap or a vein graft should be performed without hesitation to use recipient veins in a better condition. PMID- 21629067 TI - Evaluation of prophylactic anticoagulation, deep venous thrombosis, and heparin induced thrombocytopenia in 21 burn centers in Germany, Austria, and Switzerland. AB - Heparin-induced thrombocytopenia (HIT) is a life-threatening complication in intensive care settings. The timely diagnosis and management of HIT are challenging, and the incidences of HIT and deep venous thrombosis (DVT) may be related to prophylactic anticoagulation standards in burn units. We therefore evaluated, using a questionnaire, prophylactic anticoagulation, HIT management, and incidences of DVT and HIT in burn centers located in the German-speaking part of Europe. In the 21 responding burn centers, 1611 patients were treated and the overall incidences for clinically overt DVT and HIT in 2008 were 1.1% and 1.4%, respectively. Burn centers using low molecular weight heparin (LMWH) subcutaneous for all patients had a low rate of DVT (0.9%) and significantly lower rates of HIT (0.2%) relative to all other centers (P < 0.05). The highest rates of HIT (2.7%) and DVT (3.8%) were found in burn centers administering unfractionated heparin intravenous. While current HIT guidelines do not specify the administration of unfractionated heparin or LMWH for burn patients, these data warrant controlled prospective studies to confirm the advantage of LMWH administration in burn patients. PMID- 21629068 TI - Successful management of cutaneous mucormycosis by delaying debridement. AB - Mucormycosis is a frightening medical condition which has baffled clinicians all over the world. Cutaneous mucormycosis is, in particular, extremely invasive, leading to high rates of morbidity and mortality. Timely intervention with antifungal drug Amphotericin B and early radical debridement are keys for favorable outcome.Three consecutive patients died of cutaneous mucormycosis despite being treated with Amphotericin B and an early extensive debridement. With disappointing results in these patients, the treatment protocol was changed. Instead of early aggressive surgical intervention, the debridement was withheld for minimum 10 days or more, until Amphotericin B started to show its effect. Debridement was carried out conservatively after 10 days. The resultant raw area was covered with the split-thickness skin graft later.The concept of "Delay the Debridement" was efficacious in the successive 5 patients with minimum morbidity and less reconstructive requirements. Our study contradicts the popular wisdom of the necessity of early vigorous debridement. We think that the timing of debridement is one of the most important determinants of mortality.The scientific reasons for delaying the debridement have been discussed. This appears to be the first report of successful management of cutaneous mucormycosis by delaying the debridement. PMID- 21629069 TI - A postoperative analysis of perfusion dynamics in deep inferior epigastric perforator flap breast reconstruction: a noninvasive quantitative measurement of flap oxygen saturation and blood flow. AB - BACKGROUND: The blood supply of microvascular free flaps is rated as very critical, particularly in the first postoperative days. Only a timely recognition of a complication and its treatment can ensure the survival of the flap. To quantify the postoperative perfusion dynamics, we measured the oxygen supply and blood flow of the deep inferior epigastric perforator (DIEP) flaps. METHOD: A total of 23 patients undergoing DIEP flap breast reconstruction were involved in this study. Noninvasive measurements were performed with the O2C-device, which combines the determination of hemoglobin oxygenation and blood flow using white light and laser light. The first measurement was performed immediately after the flap anastomosis. Further measurements were carried out on the first, third, fifth, and seventh postoperative days. RESULTS: The preoperative value represents our baseline. With the exception of the third day, all postoperative measurements were above the baseline. On the third postoperative day, there was a decrease in the cutaneous oxygen saturation of 22% and a decrease in blood flow of 18% on average, regardless of the individual fluctuations of patient's blood pressure. CONCLUSIONS: The postoperative perfusion characteristics of the DIEP flap showed a critical drop in the cutaneous oxygen supply and blood flow on the third postoperative day. This represents a potential risk and should be considered in the postoperative management of the DIEP flap. PMID- 21629070 TI - Atypical thumb polydactyly with duplicated metacarpal bone: a report of 2 cases. AB - We report 2 cases of patients with thumb polydactyly that were difficult to classify into either Wassel type VI with a remarkable hypoplastic metacarpal or Wassel type VII (triphalangeal thumb). The radial digit was functional but hypoplastic, and the ulnar digit presented a dominant appearance but no active motion. The operative findings revealed that the radial and ulnar MP joints were completely independent and the metacarpal bones were separated, simulating Wassel type VI thumb polydactyly. In both cases, the ulnar digit was transported to the top of the radial metacarpal, accompanied by tendon transfer from the radial to the ulnar digit and rotation flap for web plasty. One year after surgery, the postoperative results showed good appearance and function of the preserved ulnar thumb in both cases. PMID- 21629072 TI - Subcaruncular approach for the reconstruction of blowout fractures of medial orbital walls. AB - To obtain a wide and clean operative field for anatomic reconstruction of medial orbital blowout fractures, an alternative method called the subcaruncular approach has been performed. Between March 2008 and June 2010, this method was applied to the orbits of 41 patients with isolated pure medial orbital blowout fractures. A medial half conjunctival incision was made and extended meticulously to the subcaruncular area through preseptal plane under direct vision. Pre- and postoperative computed tomographic scans and ophthalmic examinations were performed, and clinical results were assessed. Postoperatively, computed tomographic scans revealed anatomic reduction of orbital soft tissues and the reconstructed medial orbital wall to be in proper position in all cases, and diplopia and eyeball motility limitation were resolved in most patients. There were no complications except severe chemosis in one case. We believe that this method can be a useful alternative option for the anatomic reconstruction of medial orbital blowout fractures. PMID- 21629073 TI - Reconstruction of fingertip defects with great toe pulp grafts. AB - Various methods have been used to treat fingertip defects that are caused when distal parts are amputated. In this study, we used the pulp graft harvested from the lateral aspect of the great toe. Between September 2004 and August 2006, the great toe pulp graft were performed on 16 patients. The ages of patients ranged from 4 to 58 years. The average follow-up duration was 28 months. Complete graft take was observed in 13 of 16 patients. Partial necrosis was observed in 2 patients, and total necrosis in 1 patient. The pulp graft was painless and the color and texture of the graft were similar to the adjacent skin. Semmes Weinstein monofilament and 2-point discrimination tests showed good recovery of fingertip sense. The scar of the fingertip was assessed by Vancouver Scar Scale and Cold intolerance by Visual Analog Scale and the results were satisfactory and also improved with time. The recipient site wasclosed with little scar. There were no gait disturbances. To conclude, the great toe pulp graft can provide soft tissue and sensory recovery in fingertip defects. PMID- 21629074 TI - Extramammary Paget's disease: a novel approach to treatment using a modification of peripheral Mohs micrographic surgery. AB - Extramammary Paget's disease is a rare intraepithelial adenocarcinoma typified histologically by the presence of Paget cells. Treatment has historically been surgical, with wide excision being the standard of care. However, due to clinically indeterminate margins and diffuse spread, local recurrence rates remain high. Mohs micrographic surgery has been proposed to improve the rate of local recurrence. Application of Mohs technique to treat extramammary Paget's disease can be difficult because of the large size of these lesions. Reported excisions either involved lengthy procedures or peripheral Mohs modification. The peripheral technique does not evaluate the depth of the central lesion, yet prognosis and lymph node involvement are directly related to the degree of vertical invasion. In this study, we discuss our experience with extramammary Paget's disease, along with a novel approach to treatment by using a modification of peripheral Mohs micrographic surgery that incorporates histologic analysis of the central specimen's depth. PMID- 21629075 TI - Angiographic delay: a viable alternative to surgical delay. AB - BACKGROUND: Selective embolization of the inferior epigastric arteries can serve as a method for transverse rectus abdominis musculocutaneous (TRAM) flap delay. The purpose of this study was to determine whether delay by selective arterial embolization is comparable to traditionally surgically delayed TRAM flaps as reported in the literature, in terms of skin and fat necrosis, and to examine whether certain risk factors play a role in TRAM flap fat necrosis despite angiographic delay. METHODS: Retrospective chart review was performed for 88 consecutive patients who underwent unilateral TRAM flap breast reconstruction after selective embolization of bilateral inferior epigastric arteries. RESULTS: Between 1997 and 2009, 88 pedicled TRAM flaps were performed for breast reconstruction in women with a mean age of 49.7 years. No patients had flap skin necrosis or total flap loss. In all, 13.6% patients had TRAM flap fat necrosis. Two patients in the TRAM fat necrosis group (16.7%) had a positive history of smoking, which was a statistically significant risk factor for necrosis (P = 0.048). CONCLUSIONS: Outcomes of pedicled TRAM flaps delayed by selective arterial embolization are comparable to historical controls of those delayed by traditional surgical means (ligation of artery and vein) and better than nondelayed flaps. Smoking remains a significant risk factor for TRAM flap fat necrosis despite the benefit of delay. PMID- 21629076 TI - Usability of the middle temporal vein as a recipient vessel for free tissue transfer in skull-base reconstruction. AB - In skull-base reconstruction, finding appropriate recipient veins is difficult if the patient has undergone previous craniotomy or requires multiple drainage vessels, because of postoperative scarring and the limited number of veins in the temporal region. Therefore, we have used the middle temporal vein (MTV) to overcome these problems. We examined the presence and caliber of the MTV in 12 patients undergoing skull-base reconstruction. The MTV was identified in all cases and used for reconstruction, alone or with the superficial temporal vein (STV), in 8 patients without postoperative flap loss and was only measured in 4 patients. The caliber of the MTV was significantly greater than that of the STV, and there was no significant difference between the usage of the MTV and STV in the length of reconstruction time. Therefore, the MTV is of an appropriate caliber, and its use does not require complicated procedures as alternative recipient vessels. PMID- 21629077 TI - How to reconstruct a natural and deep umbilicus: three methods of umbilicoplasty for five types of umbilical deformities. AB - Conventional methods of umbilicoplasty using V-Y advancement flap often result in unnatural wide or shallow umbilical depressions facing upward or downward. Moreover, although the umbilical deformities have many variations, no report has described the selection of an umbilicoplasty method for types of umbilical deformity. To resolve these problems, we devised 3 methods of umbilicoplasty. In this report, we classified all kinds of umbilical deformities into 5 types, and studied the most suitable method for each type of umbilical deformity. METHOD: The umbilical deformities are classified into Type 0: the defect of umbilicus; Type I, the low-grade protrusion; Type II, the high-grade protrusion with wide base; Type III, the high-grade protrusion with narrow base; and Type IV, the protrusion in depression. The most suitable method among our 3 methods was adapted to each type. Method 1 with a S-shaped skin incision was adapted to Type 0 and I, Method 2 with fan-style flaps was adapted to Type II, and Method 3 with dividing the umbilical protrusion was adapted to Type III and IV. RESULTS: Sixty three patients (10 cases of Type 0, 31 cases of Type I, 10 cases of Type II, 5 cases of Type III, and 7 cases of Type IV) underwent umbilicoplasty using the suitable method, and all were well corrected. CONCLUSIONS: Using the best choice among our 3 methods, it is easy to create a natural, vertically long and deep umbilical depression without conspicuous scars in all types of umbilical deformities. PMID- 21629078 TI - Improving the reliability of the distally based posterior tibial artery adipofascial flap with the great saphenous venoneural network. AB - The distally based posterior tibial adipofascial flap is a useful option for distal leg and ankle coverage. Traditionally, its dimension is constrained by a length-to-width ratio of 2:1. We have increased this ratio to 4:1 by including the great saphenous vein and saphenous nerve with the flap. These structures with their venoneural network, and their connections with posterior tibial artery perforators, form the vascular axis of the flap. In our series of 21 flaps, 70% (n = 15) had ratios of 3:1 or higher. In remaining 30% (n = 6), the ratio was 2:1, as the defects were more proximal. Complications include one complete flap loss due to compression from tunneling and one superficial tip necrosis. Postoperatively, we performed wet-to-wet dressings, phlebotomy, and delayed skin grafting to optimize flap survival. Of 21 flaps, 20 were healthy after an average follow-up of 24 months. PMID- 21629079 TI - Dealing with the overprojecting pinched tip. AB - OBJECTIVE: When reshaping the pinched and/or pointed nasal tip, the use of cartilage grafting and internal sutures has been proven to be an excellent tool when the skin is not too thin. A new technique in tip aesthetics is presented. Its surgical applications are discussed. STUDY DESIGN: A tip reshaping without cartilage grafting or internal stitching was performed on 355 patients with thin skin; 317 were female and 38 were male. The follow-up period ranged from 6 months to several years, with an average of 16 months. METHODS: The closed approach was used. Parallel incisions on the alar cartilage have been made and alternate spicules of cartilage have been then excised to reduce the length of the arc of the cartilage and to reshape the tip. RESULTS: The technique was effective in achieving aesthetic norms. CONCLUSIONS: When dealing with a very thin skin, the pinched or pointed tip may sometimes need a different approach. The use of cartilage crafting or internal stitching for the reshaping of the tip, in those cases, is not obligatory. PMID- 21629080 TI - Island volar advancement flap for reconstruction of thumb defects. AB - BACKGROUND: The volar advancement flap of the thumb described by Moberg has been used for pulp defects up to 1 to 1.5 cm since its mobility is quite limited due to the stiff nature and fibrous connections of the regional skin with underlying structures. There have been several attempts to increase the mobility of this useful flap by adding V-Y and Z-plasty concepts into the technique. These modifications could provide a slight increase in the flap advancement and achieve closure of defects up to 2 cm in length at best. METHODS: In this study, we describe the island volar advancement flap of the thumb in which all attachments, except the neurovascular pedicle of the flap, were divided to provide maximum mobility and advancement. This technique was used in 12 patients for reconstruction of traumatic defects of the distal thumb up to 3 to 3.5 cm in length. RESULTS: The flaps healed uneventfully in all patients. There was no patient with flap failure. All patients were followed up for 2 to 6 years (4 years on an average). Our results revealed sensible and durable skin coverage with maximum preservation of the thumb length. CONCLUSIONS: The island volar advancement flap seems to be a safe and useful procedure for thumb reconstruction. Providing a 1.5 cm of extra flap advancement, this new procedure enables us 1-stage closure of considerably large defects and is a critical achievement in thumb reconstruction. PMID- 21629081 TI - Surgical reconstruction of auricular relapsing polychondritis. AB - Relapsing polychondritis (RP) is a multiple system disease of unknown cause characterized by episodic inflammation of cartilage and potentially progressive degeneration of cartilaginous tissue. Solitary auricular cartilage involvement without any inflammatory symptoms is very uncommon and surgical treatment is rarely recommended. We present a 53-year-old Chinese male patient who had undergone a mass resection on the right ear in December 2008 and 1 year later had developed another mass at the same spot. A mass resection and reconstructive surgery were performed and the pathologic result of the mass was auricular RP. Although RP is rarely seen by plastic surgeons, we should have sufficient information about this disease. This case demonstrates a simple and straight surgical approach to obtain an excellent ear appearance. PMID- 21629082 TI - Latissimus dorsi donor-site morbidity: the combination of quilting and fibrin sealant reduce length of drain placement and seroma rate. AB - Seroma formation has been shown to be a multifactorial process in part due to dead space and the formation of raw surfaces, which produce large quantities of serous exudate. The purpose of this study is to evaluate the effect of quilting/progressive tension sutures (to reduce dead space) and fibrin sealant (to seal the raw surface) in combination on the seroma rate and length of drain placement in patients undergoing latissimus dorsi breast reconstruction. A retrospective review of 43 patients undergoing latissimus dorsi flap breast reconstruction was performed. The patients were divided into 2 groups: quilting sutures alone versus those with quilting sutures and fibrin sealant. Data regarding age, body mass index, smoking history, timing of reconstruction, type of breast surgery, radiation history, complications, length of drain placement, use of fibrin glue, and use of quilting/progressive tension sutures were collected for each patient. Results were analyzed statistically using unpaired t tests (P < 0.05). The quilting group included 19 patients with 24 donor sites. The mean drain placement duration was 21.5 days (range, 9-69 days). One patient in 19 developed a seroma, which was treated and resolved with aspiration. The seroma rate for the quilting only group was 5%. The quilting and sealant group included 23 patients with 26 donor sites. The quilting and sealant group had a mean duration of drain placement of 13.9 days (range, 6-38 days). This was a statistically reduced length of drainage (P = 0.04) compared with quilting only. The quilting and sealant group had 1 patient in 23 develop a seroma with a rate of 4% which compared with quilting only was not statistically significant (P = 0.4). The combination of quilting sutures and fibrin sealant directed at the 2 main mechanisms of seroma formation, (dead space and serous exudate, respectively) can decrease the duration of postoperative drain placement and does maintain low seroma rates. PMID- 21629083 TI - Achieving an optimal cosmetic result with excision of lesions on the face. AB - This article offers an approach to excising lesions on the face considering a simple excision and closure by contraction, excision as an ellipse and primary closure, flap cover, and skin graft. However, the article concentrates on the geometry and mechanics of flap design. The mechanics of the 3 simple flaps (advancement, transposition, and rotation) are initially examined. Additionally, commonly used flaps, which are variations of the above, are also examined. Understanding flap geometry and design is the essence of all pedicle flaps, and will achieve an optimal cosmetic result. PMID- 21629084 TI - Aesthetic subunit of the breast: an analysis of women's preference and clinical implications. AB - Patient satisfaction in breast surgery is dependent on achieving a balance among all aesthetic subunits. The purpose of this study is to identify which subunit of the breast women consider important and correlate this clinically to improve patient satisfaction following breast surgery. A total of 313 subjects (ages, 20 80) were surveyed using a 25-point survey instrument collected via a telemedicine form. The data was analyzed to determine clinical significance. Of the subjects, 63% selected the upper inner quadrant as the most important subunit. Furthermore, 66% of the subjects indicated defects located in this region would lead them to seek operative intervention and this was consistent for all subgroups. Based on these results, defects in the upper inner quadrant of the breast are more likely to cause patient dissatisfaction. Patient outcomes following surgery can be enhanced by restoring volume and minimizing scars in this upper medial subunit of the breast. PMID- 21629085 TI - Neglected nonunion of phalangeal neck fractures of the thumb in children: the outcome of delayed bone grafting in adulthood. AB - Over a 12-year period, the author treated a total of 5 adults (mean age, 23 years) with neglected nonunion of phalangeal neck fractures of the thumb that were sustained in early childhood. Cosmetically, the affected thumb was shorter and smaller than the contralateral thumb. The thumb tip was flail and thumb pinch was weak. X-rays showed a nonunited phalangeal neck fracture with no radiologic evidence of avascular necrosis of the phalangeal head. All patients underwent iliac crest bone grafting. Bone union was obtained in all patients. At final follow-up (mean, 9 months), all patients were satisfied with the cosmetic appearance of the thumb. The thumb length increased by an average of 6 mm (range, 5-8 mm). Pinch improved in the range of 69% to 87% of the power of the contralateral thumb. However, there was restricted range of motion of the interphalangeal joint (mean range of motion of 10 degree only). It was concluded that delayed bone grafting of neglected nonunions of pediatric phalangeal neck fractures of the thumb is a worthwhile procedure and has a high satisfaction rate. PMID- 21629086 TI - Autologous dermal graft in breast reconstruction. AB - INTRODUCTION: The role of allogenic dermis is well established in reconstructive breast surgery, where it acts as a hammock between the detached inferior border of pectoralis major and the inframammary fold. METHOD: The study reports on the outcome of 19 women (21 breasts as 2 were bilateral) in which autologous dermis was used rather than allogenic material. The autologous dermis was harvested from the abdomen, as a miniabdominoplasty in 15 patients, and from the contralateral breast in 4 patients having simultaneous breast reduction/mastopexy. In all, 15 women underwent immediate reconstruction (7 with tissue expanders, 8 with immediate prosthesis), whereas 4 patients underwent delayed reconstruction (with expanders). RESULTS: Two patients developed infection requiring explantation. There were no abdominal complications after the miniabdominoplasty. The coverage of the prosthesis or tissue expander by the dermal graft was achieved on average in three-quarters of cases (range, just under two-thirds to 100%). The mean follow-up was 17 months (range, 6-36 months). CONCLUSIONS: Autologous dermis is a useful alternative to allogenic dermis. It is cheap and readily available. In patients having a contralateral breast reduction, there is no donor site. The risk of complications is not dissimilar to allogenic dermis. PMID- 21629087 TI - Postural variations after breast reduction: introduction of a new technique to achieve an objective analysis. AB - The authors performed a prospective study to evaluate postural variations arising after reduction mammaplasty, objectively recorded by means of static stabilometry. Thirty consecutive patients affected by various degrees of breast hypertrophy were enrolled in the study. Obesity, orthopedic, and otoneurologic diseases were the exclusion criteria. Postural function was evaluated with static stabilometry preoperatively and 1 and 6 months postoperatively. Patients were grouped according to age, sternal notch-nipple distance, and glandular resection and general and subgroup statistical analysis using Student t test for paired samples were performed. Statistically significant differences were found only after 6 months (P = 0.026). In the subgroup analysis, statistically significant differences were found for the age group >35 years (P = 0.0237) and for the sternal notch-nipple distance >30 cm (P = 0.0320). Stabilometry objectively demonstrated postural improvement perceived following breast reduction. Age and ptosis degree appeared to influence posture more than the entity of resection. PMID- 21629088 TI - Novel strategies for managing infantile hemangiomas: a review. AB - Complicated infantile hemangiomas require early intervention. With the advent of an improved understanding of the pathophysiology of the condition, and the serendipitous discovery of the effects of propranolol, available treatment options have expanded from the traditional use of steroids. This article reviews the current literature on the treatment of complicated hemangiomas with steroids, interferon, vincristine, bleomycin, and propranolol. The likely mechanism of action, indications, recommended treatment regimes, and complications are discussed. PMID- 21629089 TI - Freestyle propeller flaps to reconstruct defects of the posterior trunk: a simple approach to a difficult problem. AB - The reconstruction of complex defects of the back remains difficult. However, a local skin flap alone based on a competent single perforator can have a large skin territory with an efficient arc of rotation as a propeller flap to cover many back wounds, while the donor site can still be closed primarily. Eleven patients with defect sizes ranging from 36 to 264 cm(2) underwent coverage of posterior trunk defects using propeller flap (rotated perforator flaps), and all the donor sites were primarily closed. The average number of perforators within the flap was 1.4. All flaps healed uneventfully except for 1 case in which congestion was noted; leech was applied and the flap healed well. Radiation therapy was done in 1 case and was uneventful. The freestyle perforator propeller flap can be a reliable and a simpler solution to reconstruct a difficult defect while achieving minimal donor site morbidity. PMID- 21629090 TI - Long-term Outcome After Surgical Treatment of Lipedema. AB - Lipedema is a condition characterized by abnormal deposition of adipose tissue in the lower extremities leading to circumferential bilateral lower extremity enlargement typically seen extending from the hips to the ankles. Diagnosis of the condition is often challenging, and patients frequently undergo a variety of unsuccessful therapies before receiving the proper diagnosis and appropriate management. Patients may experience pain and aching in the lower extremity in addition to distress from the cosmetic appearance of their legs and the resistance of the fatty changes to diet and exercise. We report a case of a patient with lipedema who was treated with suction-assisted lipectomy and use of compression garments, with successful treatment of the lipodystrophy and maintenance of improved aesthetic results at 4-year postoperative follow-up. PMID- 21629091 TI - Growing trend of China's contribution to the field of plastic and reconstructive surgery: a 10-year study of the literature. AB - BACKGROUND: In the past decade, plastic and reconstructive surgery in China has achieved great advances. However, the scientific publications in plastic and reconstructive surgery in the major regions of China-Mainland, Hong Kong, and Taiwan are unknown. We therefore intended to reveal the contribution of articles from Chinese authors to the field of plastic and reconstructive surgery. METHODS: Articles published in the 6 journals in plastic and reconstructive surgery originating from Mainland China, Hong Kong, and Taiwan in 2000 to 2009 were retrieved from the PubMed database and Journal Citation Reports. The number of total articles, 5-year impact factors (5 years-IF), and citations were tabulated to assess both the quantity and quality of research arising from China. RESULTS: From 2000 to 2009, there were 568 articles from China, including 225 from Mainland China, 317 from Taiwan, and 26 from Hong Kong. The annual total numbers of articles from the 3 Chinese regions increased gradually between 2000 and 2009 (from 40 to 100). From 2007, the number of articles published from Mainland China exceeded Taiwan. Taiwan had the highest accumulated 5 years-IFs, average 5 years IF, total citations, and average citations of each article. Plastic and Reconstructive Surgery was the most popular journal in China. CONCLUSIONS: The number of articles published from Chinese authors increased markedly from 2000 to 2009, particularly from Mainland China, since 2004. PMID- 21629092 TI - Incorporation of a preexisting scar in the star-flap technique for nipple reconstruction. AB - BACKGROUND: This report describes the authors' currently favored method of nipple reconstruction in cases of a pre-existing scar on the breast mound that passes through the intended site of nipple reconstruction. METHODS: The star flap technique incorporating a previous scar is used at least 3 months after satisfactory implant-based breast reconstruction. If the intended site of nipple reconstruction bisects through a vertical mastectomy scar, the star flap is designed as medially or laterally based with its 2 lateral limbs lying adjacent to the scar and the transverse central component incorporating the vertical scar. On the other hand, if the selected site of nipple reconstruction passes into a horizontal scar, the flap is positioned as inferiorly or superiorly based along the scar with its 2 lateral limbs adjacent to the scar and vertical central limb including the scar. Upon ensuring viability, the limbs are inset for nipple reconstruction. The flap donor wounds of the lateral limbs are closed primarily along the previous scar, and all of the scars including that of the central limb are maintained within the area of the intended areola. RESULTS: The procedure was used 16 times in 24 patients who have undergone bilateral implant-based breast reconstruction. All flaps were viable. One nipple reconstruction had an early complication of partial flap loss in the central flap that subsequently healed with wound care. CONCLUSIONS: It is possible to use a mastectomy scar in the star flap design for nipple reconstruction with no significant vascular compromise. This novel design enables positioning of the nipple in an optimal location on the breast mound in relation with the scar, use of existing scar, and inclusion of extra skin and subcutaneous tissue within the central limb. Nipple projection and volume have been satisfactory and consistently maintained. PMID- 21629093 TI - Venous malformation: risk of progression during childhood and adolescence. AB - Venous malformations (VMs) can cause significant morbidity, particularly because they enlarge over time and become symptomatic. The purpose of this study was to determine the natural progression of VMs to facilitate patient counseling, gain insight into pathophysiology, and guide therapy. Our Vascular Anomalies Center database was reviewed for patients with cutaneous and soft-tissue VMs. Predictive variables were age, gender, location, pregnancy, and size. The outcome variable was natural progression of the malformation defined by expansion of the lesion or the onset/worsening of symptoms. The study included 614 patients. Children had a 26.1% risk of progression prior to adolescence, 74.9% before adulthood, and 93.2% over their lifetime. Progression was more likely in adolescence (60.9%) than in childhood (22.5%); the relative risk was 2.6 (95% confidence interval, 2.1-3.2) (P = 0.0001). Diffuse VMs progressed more often than localized lesions (P = 0.002); extremity and trunk VMs worsened more frequently than head/neck lesions (P = 0.03). VMs have a higher risk of progression in adolescents than in children; pubertal hormones may contribute to expansion. Because of their high rate of progression, early treatment of asymptomatic VMs should be considered. PMID- 21629094 TI - Multiple closed avulsions of flexor tendons of the hand caused by a firecracker blast. AB - Closed avulsion, in the absence of pathology, of both flexor digitorum profundus and superficialis tendons from the same finger is uncommon. We report a multiple closed avulsion of long fingers flexor tendons, associated with pulleys rupture in the left hand of a healthy, 16-year-old boy. The lesion was caused by a firecracker blast which the patient had tried to hide. No injuries were found in the volar or dorsal areas of the hand. All affected flexor tendons were reinserted; in addition, second and third digits fourth annular pulleys were resutured to prevent posterior bowstringing. At present, the patient has completely recovered his range of motion and has obtained his driver license. PMID- 21629095 TI - The effect of obesity on early outcomes in adolescents undergoing reduction mammaplasty. AB - It is not known whether obesity portends poorer outcomes following reduction mammaplasty in adolescent macromastia patients. We review symptoms in obese and nonobese adolescent macromastia patients and describe early outcomes following reduction mammaplasty. Demographics, operative details, and postoperative follow up data were collected on 67 patients seen at our institution between 1997 and 2008. Variables were compared using 2-sample t tests or Pearson chi/Fisher exact tests. Mean age at surgery was 17.1 +/- 1.6 years. Mean body mass index was 27.9 +/- 4.5 kg/m, and 32.8% were obese. Thirty-four patients (50.7%) experienced minor complications; 1 patient experienced a major complication. Of patients with complications, obese patients reported a greater number than nonobese patients (P = 0.013). There were no differences in the type of complication or self-reported satisfaction between obese and nonobese patients 34.4 +/- 25.7 weeks after surgery. Our findings suggest that reduction mammaplasty is well-tolerated in obese and nonobese adolescents with macromastia and that obesity is not an absolute contraindication to reduction mammaplasty in adolescents. PMID- 21629096 TI - The effect of scrotal reconstruction with skin flaps and skin grafts on testicular function. AB - Due to its unique composition, the reconstruction of scrotal skin defects is a major clinical challenge. This study was designed to evaluate the effects of scrotal reconstruction, using skin grafts and skin flaps, on spermatogenesis. In Group 1, the rats did not undergo surgery and were used as controls. In Group 2, after removal of all of the scrotal skin to expose the testicles, the defect was repaired using a skin flap from the right groin region. In Group 3, the reconstruction was achieved using skin grafts. All the rats were killed at 2 months postoperatively and evaluated. The mean wet weights of the testicles in the control group were significantly higher compared with that of the graft group. The mean height of the germinal epithelium was significantly greater in the control and flap groups compared with that of the graft group. The Johnsen score for spermatogenesis in the control group was higher than that in the graft group. The use of flaps resulted in testicular function that was comparable to that of the control group, whereas the use of grafts resulted in diminished testicular function. Therefore, we suggest that flaps may be the first choice for scrotal reconstruction. PMID- 21629097 TI - "Uro-abdominoplasty": an adaptation of abdominal contouring for revision of complicated urostomies. AB - BACKGROUND: The aim of this study was to describe the indications, surgical technique and outcomes of abdominoplasty as a novel tool for revising complicated urostomies. PATIENTS AND METHODS: Four patients (3 female, 1 male; mean body mass index = 32 kg/m; mean age = 56 years) who underwent abdominoplasty for urostomy revision 2007-2009 were identified. Ileal conduits had been performed following ablative or diversion surgery for cervical carcinoma, bladder carcinoma, interstitial cystitis, and neuropathic bladder. A postal questionnaire was used to establish pre- and postabdominoplasty stoma function. RESULTS: Patients were referred to the reconstructive team with problems fitting their urostomy appliance leading to urinary leakage, skin irritation, and social embarrassment. Uro-abdominoplasty indications included multiple abdominal scars (n = 2), large abdominal apron (n = 4), and deep skin creases (n = 2). Three patients had undergone previous failed urostomy repositioning or peristomal liposuction. The joint plastic surgical-urological operations lasted a mean of 3 hours, with no major postoperative complications. Patients were discharged 8 days later. Of 4 patients, 3 reported improved appliance fitting and reduced urinary leakage (>50%) and the remaining patient had intermittent leakage due to a persistent abdominal fold superiorly, and has since undergone reverse abdominoplasty. Two patients complained of long-term lower abdominal numbness, but all 4 were satisfied with the aesthetic improvement. CONCLUSIONS: Abdominoplasty has been successfully used in our center for the purpose of improving urostomy dysfunction of intractable mechanical leakage by creating a flatter surface for appliance fitting. Uro-abdominoplasty widens the reconstructive repertoire of plastic surgeons and can be considered in those who have exhausted conservative or simpler surgical solutions. PMID- 21629098 TI - Endoscopic transnasal approach for the treatment of isolated medial orbital blow out fractures: a prospective study of preoperative and postoperative orbital volume change. AB - Endoscopic transnasal reduction is a safe and effective technique for the treatment of blow-out fractures of the medial orbital wall. However, because this approach does not use rigid permanent material for reconstruction of the fractured medial orbital wall, some degree of herniation of the orbital contents may occur after the intraethmoidal packing material is removed. The purpose of this study was to evaluate the change in orbital volume in patients with medial orbital wall fractures treated through an endoscopic transnasal approach. This study was a prospective analysis that includes 20 patients who underwent endoscopic transnasal reduction of medial orbital wall fractures between April 2007 and December 2008. Computer-assisted orbital volume measurements were made using axial computed tomography. The mean (standard deviation [SD]) volume increase was 2.00 (0.92) cm(3) and the mean (SD) dimension of the fractured orbital wall was 2.76 (0.83) cm(2). After endoscopic surgery, an average (SD) volume decrease of 2.15 (0.91) cm(3) was achieved with ethmoid sinus packing. After removal of the packing materials, 1.14 (0.78) cm(3) increase of the orbital volume was observed. The dimension of the orbital wall fracture significantly correlated with the increased preoperative orbital volume (P = 0.002, r = 0.609); the preoperative increase in the orbital volume also significantly correlated with volume relapse after removal of the packing (P = 0.023, r = 0.452). These findings suggest that in broad orbital wall fractures, reconstruction of the orbital wall by rigid materials or prolongation of the packing period should be considered, because orbital volume can increase again after packing removal, and may thus lead to postoperative complications. PMID- 21629099 TI - Outcome study of the surgical management of panniculitis. AB - Patients with panniculus morbidus have an abdominal panniculus that becomes a pathologic entity, associated with the development of candidal intertrigo, dermatitis, lymphedema, and ischemic panniculitis. Panniculectomy is a standard treatment for this problem. The objective of this study was to determine risk factors for complications associated with panniculectomy surgery to lower the complication rate. We performed a retrospective chart review of patients who underwent panniculectomy between 1999 and 2007 by looking at data related to surgical complications, comorbidities, age, and gender. In 563 patients, we recorded the incidence of the following complications: wound-related (infection, dehiscence, and/or necrosis), hematoma/seroma, respiratory distress, blood transfusions, deep venous thrombosis or pulmonary embolism, and death. Overall, 34.3% of patients suffered at least 1 complication. In patients with wound complications specifically, there was a significantly higher body mass index versus those with no wound complications (43.7% vs. 30.7%, P < 0.0001). Smokers also had a higher rate of wound complications (40.5% vs. 19.5%, P < 0.0001). PMID- 21629100 TI - Modified Rives-Stoppa technique for repair of complex incisional hernias in 59 patients. AB - Incisional hernias develop in 2% to 11% of patients who undergo laparotomy. Prosthetic mesh repair provides more strength, tension-free closure, and decreased recurrence rates as compared to primary tissue repairs. Complications fistula formation, adhesions, skin erosion, and seroma/abscess formation-however, include increased rates of infection, sometimes requiring complete mesh removal. The Rives-Stoppa repair for complex incisional hernias confers the benefits of prosthetic repair and lower recurrence rates, but decreases certain complications by preventing direct mesh contact with the bowel. A total of 89 consecutive patients (mean age, 58.1) underwent a modified Rives-Stoppa repair for purposes of this review, all the patients who lost to follow-up before 6 months postoperatively were excluded from the study. Of the remaining 59 patients, 32.2% (n = 19) had expanded polytetrafluoroethylene mesh, and 67.8% (n = 40) had polypropylene mesh. Average range of follow-up was 40.0 months. Hernia recurred in 1 patient (1.7%). Infection requiring explantation of the prosthesis occurred in 3 patients (5.1%). The Rives-Stoppa repair is reportedly the best open technique for complex incisional hernias with comparatively lower recurrence rates. Additionally, patients with inflammatory bowel disease (64.4% of our series), who often require later reoperation for their primary disease, may benefit from this technique of herniorrhaphy where no interface exists between intrabdominal contents and the prosthesis. This lack of interface decreases intrabdominal adhesions and facilitates re-entry if future surgery is needed for inflammatory bowel disease. PMID- 21629101 TI - Assessment of configuration of thoracic duct using magnetic resonance thoracic ductography in idiopathic lymphedema. AB - The structure and the function of the peripheral lymph channels have been investigated, but the thoracic duct has not. This study used magnetic resonance thoracic ductography for 2 patients with idiopathic lymphedema to evaluate the configuration of the thoracic duct in these patients. Anomalies of the thoracic duct were detected in both cases. This result suggests that deformity of the thoracic duct is one of the causes of idiopathic lymphedema. Characterization of the etiology could lead to a breakthrough in resolving the occurrence of idiopathic lymphedema and developing a treatment procedure for it. In addition, visualization of the thoracic duct may assist in selecting the optimal therapy for each idiopathic lymphedema patient. PMID- 21629102 TI - Effect of electroporation-mediated transfecting recombinant plasmid pIRES-hBMP2 hVEGF165 on mandibular distraction osteogenesis. AB - Distraction osteogenesis requires a long consolidation period and has a low but real failure rate. Bone morphogenetic proteins (BMPs) accelerate bone deposition in fractures and critical-sized bone defects. Vascular endothelial growth factor (VEGF) is a promising reagent for inducing angiogenesis, and is an essential coordinator of extracellular matrix remodeling, angiogenesis, and bone formation in the growth plate. However, their effects on mandibular distraction osteogenesis are unknown. We investigated the effect of local delivery of plasmid pIRES-hBMP-2-hVEGF165 into a distraction area by electroporation-mediated approach.A New Zealand rabbit model were used. Activation of the device was commenced after 3 days of latency period and proceeded at the rate of 0.8 mm per day for 7 days. After the completion of activation, the rabbits were randomly divided into 5 groups: group A: recombinant plasmid 2 MUg (0.1 MUg/MUL) pIRES hVEGF165-hBMP2 was injected into the distraction area after the completion of activation; group B: recombinant plasmid pIRES-hBMP2 was injected into the distraction area; group C: recombinant plasmid pIRES-hVEGF165 was injected into the distraction area; group D: pIRES was injected into the distraction area, and group E: normal saline was injected into the distraction area. After injection every group used electroporation. Subsequently, the rabbits were examined by quantitative computed tomography, mechanical testing, and histomorphometric analysis.BMD of newly formed bone of the distraction area in groups A, B, and C were remarkably higher than those of groups D and E at different times (P < 0.001). At 4 and 8 weeks of consolidation, the crushing strength of 3 points of the newly formed bone in group A was remarkably higher than those of groups B, C, D, and E (P < 0.01). The results demonstrated statistically remarkable increase in regenerated bone in the gene-transfected groups.Electroporation-mediated transfecting recombinant plasmid pIRES-hVEGF165-hBMP2 could produce a satisfactory proceeding of osteogenesis and calcification, which surpassed that of the control group. This finding indicates that a combination of VEGF and BMP may make osteogenesis and angiogenesis appear at the same time. Furthermore, it may magnify the effect of single growth factor, and promote growth and reparative process of bone. PMID- 21629103 TI - Clinicopathological characteristics and surgical outcomes of divided nevus of the eyelids: a decade's experience on 73 cases. AB - Currently, there is a lack of comprehensive data regarding clinicopathologic characteristics and surgical treatment of divided nevus of the eyelids. Here clinicopathologic features and reconstructive procedures performed on 73 patients were described. We showed that the lesion was randomly located on both eyes with a predominant incidence in females. Affected regions included the following: eyelid only (n = 40, 54.8%); medial canthus involvement (n = 4, 5.5%); medial canthus, caruncle, and bulbar conjunctiva involvement (n = 5, 6.8%); lateral canthus involvement (n = 18, 24.7%); full eyelids (n = 6, 8.2%); and cheek extension (n = 9, 12.3%). Lesions presented as a pigmented macule (n = 18, 24.7%) or mass (n = 55, 75.3%). Ptosis was present in 26 (35.6%) cases leading to visual field defects (n = 9, 12.3%). The puncta were covered in 13 (17.8%) cases. Overall, surgical results were satisfactory. Pathologically, lesions were either intradermal (n = 40, 54.8%), junctional (n = 15, 20.5%), or compound (n = 18, 24.7%). Because of the lack of systemic data regarding the rare entity of divided nevus of the eyelids, this study improves our understanding of this lesion, and it should be treated with individualized reconstructive procedures. Incorporation of pediatric patient population in the future will further extend our knowledge in dealing with this disorder. PMID- 21629104 TI - Stretch test: effectiveness in identifying basal cell carcinoma borders. AB - Basal cell carcinoma (BCC) is the most common malignant tumor in the world. Total tumor ablation is the gold standard of treatment. Our aim in this study was to evaluate the effectiveness of a new clinical method, we dubbed the "stretch test," in reducing the incidence of incomplete excision of BCC. All BCCs excised between October 2001 and June 2007, by 1 plastic surgeon, served as the treatment population. Until September 2004, BCCs were excised in the traditional manner and this group served as a control. From October 2004, the stretch test was used in all patients with BCC and this group served as a study group. Overall, the incomplete excision rate was 3.7% (22/596). The rate of incompletely excised BCC until 2004 was 6.4% (16/249) and it decreased significantly (P < 0.05) from 2004 to 1.7% (6/347) in the group in which the stretch test maneuver was used. We found a significant decrease in the rate of incomplete excision of BCC when the stretch test was employed. This reduces the need for reexcision, improves patient recovery, and satisfaction, and helps reduce healthcare costs. PMID- 21629105 TI - Massive localized lymphedema: review of an emerging problem and report of a complex case in the mons pubis. AB - Massive localized lymphedema (MLL) is an emerging complication of the obesity epidemic. Caused by the obstruction of lymphatics, MLL presents as a giant swelling, with characteristic skin changes, and often lymphatic weeping. MLL has also been called "pseudosarcoma" because of its morphologic and pathologic similarity to sarcoma. Left untreated, MLL can degenerate into angiosarcoma. We present a case of MLL of the mons pubis in a 40-year-old man with a body mass index of 69. The literature is reviewed and an additional 40 cases of MLL are described. We found a female predominance of 1.6 to 1, an average weight of 421 lbs, and a 58% majority of cases in the thigh. PMID- 21629106 TI - Brain plasticity in Mobius syndrome after unilateral muscle transfer: case report and review of the literature. AB - BACKGROUND: Mobius syndrome is mainly characterized by bilateral facial palsy. Facial reanimation of these children is achieved by microsurgical techniques, namely free-gracilis muscle innervated by the masseteric nerve. Notorious commissure excursion and speech improvement are reported with such procedure. Several studies have demonstrated the presence of cortical reorganization after injury and repair of different segments of the body. Intensive training of a behaviorally relevant task is key in this process. CASE REPORT: A 4-year-old patient with complete bilateral facial palsy secondary to Mobius syndrome was operated with left hemiface free-gracilis muscle transplant innervated by the masseteric nerve and submitted for postoperative physiotherapy. Eight months later, bilateral movement was noted. CONCLUSIONS: Brain plasticity is likely to play an important role in smile restoration in patients with bilateral facial palsy. Intensive physiotherapy and psychosocial relevance of facial expression might be key in such phenomenon. PMID- 21629107 TI - Lateral intercostal artery perforator-based reversed thoracodorsal artery flap for reconstruction of a chronic radiation ulcer of the lower back wall. AB - Flaps with adequate blood supply are the best methods for covering the radiation ulcer defect. Our report is on the use of the lateral intercostal artery perforator-based reversed thoracodorsal artery (TA) flap for treatment of a patient with a large radiation ulcer on his lower back. When the flap was elevated, we could use an infrared imaging device to confirm the location of the perforators and demonstrate the communication with the TA. The communication between the main TA and the lateral intercostal artery perforator has previously not been reported in the literature in detail. We used an indocyanine green dye and infrared imaging device to seek out the perforators and their communication. Even in a small communication, we were able to use the device to check the perforators and to elevate this flap with more assurance, without having to be concerned about further radiation exposure for the patient. PMID- 21629108 TI - Identifying and managing those patients at risk for aborted mohs micrographic surgery. AB - BACKGROUND: Treatment of nonmelanoma cutaneous carcinoma with Mohs micrographic surgery (MMS) is a well-recognized therapy. However, there are infrequent times when MMS must be aborted before achieving adequately clear margins. Reasons cited by those who have aborted MMS include patient discomfort and/or concern regarding damage to deeper structures. METHODS: A retrospective cohort was created consisting of patients who were referred for reconstructive procedures following MMS during a 4-year period. Patients who experienced aborted MMS were identified from this cohort, and a chart review performed. RESULTS: The overall occurrence of aborted MMS in the community was significantly less than 1% with a cumulative frequency in this particular reconstructive cohort of 1.7%. Approximately one third of patients had persistent tumor on subsequent treatment, and approximately 14% required multiple excisional procedures to clear tumor. Risk factors for experiencing aborted MMS include histology of dermatofibroma sarcoma protuberans, significant pain during MMS, and location of tumor over a cranial nerve. Follow up failed to reveal recurrence after further treatment. CONCLUSIONS: Preoperative identification of patients at risk for aborted MMS would allow for direct referral using surgical excision, hence decreasing anxiety, morbidity, and cost. A treatment algorithm is proposed for patients who experience aborted MMS. PMID- 21629109 TI - Free radial forearm fasciocutaneous flap in the treatment of distal third tibial osteomyelitis. AB - BACKGROUND: Osteomyelitis of the lower limb, associated with soft-tissue defects, is a reconstructive challenge. Microvascular free-tissue reconstruction is an important surgical option with the superiority of free muscle and myocutaneous flaps being explored by various publications. Muscle flaps provide good quality vascularized tissue which can be contoured into defects, but their bulk can often lead to the reconstruction having a suboptimal aesthetic outcome. This result is usually most evident in the lower third of the tibia. We present our series of free radial forearm fasciocutaneous flaps for reconstruction of distal third tibial soft-tissue defects following debridement of osteomyelitic foci and bony stabilization. METHODS: A retrospective case-note review of 20 patients treated with free radial forearm fasciocutaneous flap reconstruction of distal third tibial soft-tissue defects following excision of osteomyelitis and adjacent scar tissue, and bony stabilization between January 1999 and December 2006 was conducted. RESULTS: There were 20 patients who had established osteomyelitis of the distal third of tibia following previous open fractures. The mean size of the soft-tissue defect at the time of the free fasciocutaneous flap procedure was 72.3 cm(2) and the mean bony defect was 3.4 cm. The mean duration of the procedure was 417 minutes and flap ischemia time did not extend beyond 60 minutes in any of the cases. Overall flap survival was 100% and all patients had radiologic bony union at a mean of 5.73 months. The average period of follow-up was 22.5 (range, 19-36) months, and none of the patients had a recurrence of the disease during the follow-up period. CONCLUSION: Free fasciocutaneous flap reconstruction for soft-tissue defects of the lower limb in our series was seen to be safe, reliable, and provided an aesthetic outcome with high patient satisfaction rates. We advocate the use of free fasciocutaneous flaps for small to-moderate sized defects of the distal third of the tibia after debridement for chronic osteomyelitis. Although we have used radial forearm flaps in our series, more modern free fasciocutaneous flaps with less donor-site morbidity may be preferred. PMID- 21629110 TI - Free tensor fascia lata-iliac crest osteomusculocutaneous flap for reconstruction of combined maxillectomy and orbital floor defect. AB - Reconstruction of maxillectomy with extensive orbital rim and floor excision defects is a challenging problem. The goal of reconstruction here is to provide adequate orbital support to prevent enophthalmos and diplopia as well as obturation of the palatal defect. The existing methods of the reconstruction fail to simultaneously address these 2 goals of reconstruction. A new method of reconstruction of these defects using tensor fascia lata-iliac crest flap was used in 7 cases of cancers of the maxilla, which necessitated extensive resection of the orbital floor along with the maxillectomy. The flap was raised as a muscle and bone flap in 5 cases and in 2, a skin paddle was included. The immediate and delayed outcome at 6-month follow-up was analyzed. The functional outcome with regards to the ocular position and function, palatal obturation, speech, and swallowing were recorded. The bone viability at 6 months was assessed by computed tomography scan. The flap was successful in all the 7 cases. The delayed outcome assessment showed that the orbital support was excellent with no diplopia in all the cases. The palatal defect could be covered successfully in all the cases, resulting in normal speech and swallowing. The computed tomography scan showed excellent integration of the bone. The free tensor fascia lata-iliac crest flap is a reliable and safe method of reconstruction of the orbitomaxillary defects, addressing the issues of both orbital support and palatal obturation. PMID- 21629111 TI - Faster wound healing with topical negative pressure therapy in difficult-to-heal wounds: a prospective randomized controlled trial. AB - OBJECTIVE: A randomized clinical trial was conducted to determine the effectiveness and safety of topical negative pressure therapy in patients with difficult-to-heal wounds. METHODS: A total of 24 patients were randomly assigned to either treatment with topical negative pressure therapy or treatment with conventional dressing therapy with sodium hypochlorite. The study end point was 50% reduction in wound volume. The maximum follow-up time was 6 weeks. RESULTS: The median treatment time to 50% reduction of wound volume in the topical negative pressure group was 2.0 weeks (interquartile range = 1) versus 3.5 weeks (interquartile range = 1.5) in the sodium hypochlorite group (P < 0.001). The unadjusted hazard rate ratio for the time until 50% wound volume reduction was 0.123 (P < 0.001). After adjustment for relevant baseline characteristics in a Cox proportional hazards model treatment group, membership was found as the only and statistically significant indicator for the time to 50% wound volume reduction (hazard rate ratio of 0.117 [P < 0.001]). Subgroup analysis of spinal cord injured patients with severe pressure ulcers showed similar statistically significant results as in the total wound group. CONCLUSION: Topical negative pressure resulted in almost 2 times faster wound healing than treatment with sodium hypochlorite, and is safe to use in patients with difficult-to-heal wounds. PMID- 21629112 TI - Reconstruction of fronto-orbital and nasal defects with compound epoxied maleic acrylate/hydroxyapatite implant prefabricated with a computer design program. AB - The repair of fronto-orbital nasal bone defects may be a troublesome problem to plastic surgeons. This report aims to present the results of reconstruction of fronto-orbital nasal bone defects with the prefabrication of epoxied maleic acrylate/hydroxyapatite compound (EH compound) using 3-dimensional (3D) imaging data and computer-assisted manufacturing techniques, sometimes combined with autogenous bone to repair nasal defect. Helical computed tomography data were used to create a 3D model of the patient's skull. On the basis of these data, the individual shape of the implant was created by a computer-aided design/computer aided manufacturing program. A rapid prototyping system was applied for production of the physical models. A total of 12 patients with traumatic fronto orbital nasal defects were included in this study. The patients followed up for 1 to 24 months. The satisfactory results, such as excellent symmetry, stability, and normal fronto-orbital contours were obtained for all patients. The operating time was short without any complications. The depression of the region of fronto orbital nasal bone defects always achieved an attractive or satisfactory prominence that was in balance and harmony with other facial features of all the patients. This method allows accurate fabrication of the implant. It improves the surgical techniques and reduces the risk of a second intervention, with improved aesthetic outcomes. PMID- 21629113 TI - A new skin flap method for total auricular reconstruction: extended scalp skin flap in continuity with postauricular skin flap and isolated conchal flap: four skin flaps and temporoparietal fascia flap. AB - A sufficient skin envelope of good quality as well as definite auricular framework is a prerequisite for a successful auricular reconstruction. Various surgical techniques, such as recruitment of mastoid skin, skin graft, tissue expansion, and so on, have been used to get the necessary skin for covering of the auricular framework. However, debates about the drawbacks of these techniques have continued. In this article, I report on a new skin flap method for total auricular reconstruction, which is an extended scalp skin flap in continuity with postauricular skin flap and isolated conchal flap. Between January 2009 and March 2010, a total of 20 patients underwent an auricular reconstruction using a Medpor framework (Porex Surgical, Inc, Newnan, GA) and the new skin flap method. Follow up time range was 4 to 17 months. The reconstructed ear showed no definite true hair growth except for some fine hair, which can be ignored. More favorable results such as a good color matched skin, well-formed ear convolution, no other donor site scars can now be achieved using this new method. PMID- 21629114 TI - Treatment of posttraumatic fingertip pain using a great toe pulp graft. AB - Complications of fingertip injury include pain, hyper- or dyssensitivity, cold intolerance, and fingertip atrophy. Especially in cases of soft-tissue defect or atrophy which result from crushing injury, fingertip pain often occurs when a finger touches the objects. To overcome this problem, several techniques including local flaps or free flaps were suggested. But these methods require intricate and multistaged procedures.Twelve patients who had fingertip pain with pulp atrophy were treated with pulp graft between March 2004 and March 2006. Under the local anesthesia, we made a fish-mouth incision at the most prominent portion of fingertip and elevated volar flaps. Composite tissue was harvested from the lateral aspect of great toe, and inserted between the previously elevated volar flaps. The harvested composite pulp tissue contained about 3- to 5 mm thick fat layer. Moisture dressing was performed. The visual analogue scale (VAS) was used to evaluate the degree of pain postoperatively. The follow-up period was in the range between the 12 and 24 months (average, 19 months). Pre- and postoperative differences in VAS scores were analyzed for statistical significance, using the Wilcoxon rank sum test. In addition, patients were asked about their level of satisfaction with the procedure. To evaluate the postoperative sensation of the graft, we performed the Semmes-Weinstein monofilament test, and static and dynamic 2-point discrimination test at 1 year postoperatively.The size of the graft was ranged from 276 mm (12 * 23 mm) to 750 mm (25 * 30 mm). At final follow-up review, 5 patients were very satisfied and 7 were satisfied. Atrophy of the fingertip was also improved. Fingertip pain reduced from 8.5 preoperative to 3.1 postoperative on VAS. These improvements were statistically significant. Semmes-Weinstein monofilament test was green (~2.83) in 9 patients (75%) and blue (3.22-3.61) in 3 of 12 patients (25%). Static and dynamic 2-point discrimination test results came out as 6 and 5 mm, respectively.Composite graft applied to the fingertip is a simple technique, and gives few complications. This procedure can be performed under local anesthesia and gives a fairly high degree of satisfaction to patients. We believe this method is useful for treating fingertip pain with atrophy of pulp. PMID- 21629115 TI - Agenesis of the vocal cords in a female infant with Robin sequence. PMID- 21629116 TI - Necrotic core and thin cap fibrous atheroma distribution in native coronary artery lesion-containing segments: a virtual histology intravascular ultrasound study. AB - OBJECTIVES: The aim of this study was to assess the longitudinal topographical relationships between minimal luminal area (MLA) sites and plaques with the most vulnerable characteristics using radiofrequency-based virtual histology intravascular ultrasound analysis. METHODS: We analyzed 69 native coronary artery segments with de-novo lesions (>50% stenosis) obtained from 50 patients with ischemic coronary artery disease. Maximal necrotic core (maxNC) was defined as a virtual histology intravascular ultrasound frame with the maxNC area and virtual histology-characterized thin cap fibrous atheroma was defined as a cross-section, which contained a plaque burden of more than 40%, relative necrotic core area of 10% or more, and a narrow band encircling the lumen containing relative necrotic core area of more than 10%, in three consecutive frames. RESULTS: MaxNC was present at the MLA site in only 17.4% of the segments, proximal in 52.2% (by 5.0 +/- 5.4 mm), and distal to MLA in 30.4% (by 4.0 +/- 5.1 mm). Non-MLA sites with maxNC (n=57) compared with MLA sites had reduced plaque burden (64.5 +/- 11.2% vs. 76.0 +/- 10.5%, P<0.001), increased remodeling index (1.04 +/- 0.17 vs. 0.89 +/- 0.15, P<0.001), less fibrotic tissue (47.7 +/- 13.4% vs. 54.8 +/- 13.8%, P<0.001), and higher dense calcium deposition (15.3 +/- 10.8% vs. 11.9 +/- 10.3%, P<0.001). Plaques containing maxNC and virtual histology-characterized thin cap fibrous atheroma were found in 23 of the non-MLA sites compared with two of the MLA sites (P<0.0001). CONCLUSIONS: In coronary artery segments with intermediate to-severe stenosis, plaques containing maxNC are mostly located away from the MLA site and more often comprise virtual histology-characterized thin cap fibrous atheroma. Such data may carry practical implications for coronary revascularization procedures. PMID- 21629117 TI - On sperm counts and data responsibility. PMID- 21629118 TI - Trends in sperm counts: the saga continues. PMID- 21629119 TI - Four hand injuries not to miss: avoiding pitfalls in the emergency department. AB - The clinical and radiological findings in some hand injuries can be subtle and easily misinterpreted, leading to irreversible changes and profound functional loss. The importance of early and accurate diagnosis is clear. This study looks at four such injuries, with reference to pertinent anatomy, typical mechanisms of injury, examination and radiological findings, with emphasis on avoiding pitfalls in the emergency department. PMID- 21629120 TI - Development of an acute ischemic stroke management course for hospital physicians in emergency departments and intensive care units. AB - The objective of this study was to evaluate a training course on acute ischemic stroke (AIS) for hospital physicians, part of a regional strategy on AIS patient care. The course comprised an initial self-study (e-Learning) stage and 1-day theoretical-practical course on initial AIS management for Critical Care and Emergency physicians in the Andalusian Health Service (Spain). Data were collected on regional implementation of the stroke code and intravenous thrombolysis treatment. Between 2006 and 2009, 12 courses were attended by 356 physicians from emergency (n=148) and critical care (n=208) departments in the Andalusian health system. The initial stage was failed by 46.4% of trainees; the 1-day AIS course was successfully completed by all trainees, who reported a high satisfaction level. By the end of 2009, all hospitals had adopted the stroke code and approximately 5-6% of patients with AIS received intravenous thrombolysis. This type of healthcare strategy proved effective to improve AIS care in our setting. PMID- 21629121 TI - Physical fitness affects the quality of single operator cardiocerebral resuscitation in healthcare professionals. AB - OBJECTIVE: Sustained external chest compressions during cardiocerebral resuscitation (CCR) are physically demanding. It might be hypothesized that a high cardiopulmonary exercise capacity and/or muscle strength delays the development of physical fatigue and, consequently, preserves CCR quality. We intended to assess the impact of cardiopulmonary exercise capacity and muscle strength on CCR quality. METHODS: Fifteen healthcare professionals (10 men and five women, mean age 34+/-9 years) performed a 15-min hands-on CCR session on an adult training manikin. CCR compression depth (from which CCR quality was calculated) and frequency were monitored. During CCR we assessed serial blood lactate concentrations, and provided continuous heart rate monitoring. Relationships were examined between participant characteristics, peak cardiopulmonary exercise capacity, ventilatory threshold, maximal muscle strength, muscle strength endurance and CCR quality. RESULTS: Significant univariate correlations were found between 15-min CCR quality and body height (r=0.53), ventilatory threshold (r=0.67), peak oxygen uptake capacity (r=0.54), peak cycling power output (r=0.54), and maximal isometric elbow extension strength (r=0.55) (P<0.05). CCR quality was significantly lower in females, when compared with males (P<0.05). Within different timeframes, CCR quality was mainly related to the ventilatory threshold up to the first 5 min (P<0.05), whereas CCR quality was mainly related to maximal isometric elbow extension strength after 5 min (P<0.05). CONCLUSION: In healthcare professionals, the ventilatory threshold is significantly related to CCR quality during the first few min. Healthcare professionals who are regularly involved in CCR should therefore aim to achieve/sustain a high aerobic exercise capacity. CLINICAL TRIAL REGISTRATION INFORMATION: Study registration number: ISRCTN70447230, www.controlled trials.com/ISRCTN70447230. PMID- 21629122 TI - Inverse correlation between allergy markers and Helicobacter pylori infection in children is associated with elevated levels of TGF-beta. AB - OBJECTIVES: We evaluated allergy/hypersensitivity clinical markers and their correlation with Helicobactor pylori infection in children and adults to analyze how early acquisition of H. pylori could modulate allergic disorder expression. PATIENTS AND METHODS: H. pylori presence was assessed by the rapid urease test and histology of antrum biopsies in 165 patients. Skin tests, serum IgE, and two clinical allergy questionnaires were performed. Allergy severity was operationally defined using a combined score. Findings were correlated with H. pylori status and cytotoxin-associated gene A presence in pediatric and adult patients. Transforming growth factor beta (TGF-beta) levels were measured by an enzyme-linked immunosorbent assay in serum and gastric biopsies of H. pylori (+) patients. RESULTS: H. pylori (-) children had more positive skin tests to a higher number of antigens than H. pylori (+) children (P<0.05). Operationally defined allergy inversely correlates with H. pylori infection in children, but not in adults. The percentage of H. pylori infection was lower in children with severe allergy (32.3%) compared with children with mild allergy (43.4%) or no allergy (64.3%) (P<0.05). Colonization with virulent strains (cytotoxin associated gene A+) showed a nonsignificant inverse correlation with severity of allergies in pediatric patients. H. pylori-infected children, but not adults, without allergy markers showed increased levels of TGF-beta compared with allergic children both in serum and gastric mucosa (P<0.05). CONCLUSION: There was a strong inverse correlation between allergy markers and H. pylori infection in pediatric patients associated with elevated levels of TGF-beta locally and systemically. H. pylori-associated chronic gastritis might downregulate clinical allergy expression. PMID- 21629123 TI - Personalized cancer genetics training for personalized medicine: improving community-based healthcare through a genetically literate workforce. AB - PURPOSE: To assess the impact of a multimodal interdisciplinary course on genetic cancer risk assessment and research collaboration for community-based clinicians. Clinicians are increasingly requested to conduct genetic cancer risk assessment, but many are inadequately prepared to provide these services. METHODS: A prospective analysis of 131 participants (48 physicians, 41 advanced-practice nurses, and 42 genetic counselors) from community settings across the United States. The course was delivered in three phases: distance didactic learning, face-to-face training, and 12 months of web-based professional development activities to support integration of skills into practice. Cancer genetics knowledge, skills, professional self-efficacy, and practice changes were measured at baseline, immediate, and 14 months postcourse. RESULTS: Knowledge, skills, and self-efficacy scores were significantly different between practice disciplines; however, postscores increased significantly overall and for each discipline (P < 0.001). Fourteen-month practice outcomes reflect significant increases in provision of genetic cancer risk assessment services (P = 0.018), dissemination of cancer prevention information (P = 0.005) and high-risk screening recommendations (P = 0.004) to patients, patient enrollment in research (P = 0.013), and educational outreach about genetic cancer risk assessment (P = 0.003). CONCLUSIONS: Results support the efficacy of the multimodal course as a tool to develop a genetically literate workforce. Sustained alumni participation in web-based professional development activities has evolved into a distance mediated community of practice in clinical cancer genetics, modeling the lifelong learning goals envisioned by leading continuing medical education stakeholders. PMID- 21629124 TI - Quality of life, disability, and body mass index are related in obese patients. AB - The aim of this study is to analyze the relationship between health-related quality of life (QoL), disability, and degree of obesity. Adult obese patients (BMI>30) were consecutively enrolled in this cross-sectional observational study. The WHO Disability Assessment Schedule (WHO-DAS II) and the short version of the impact of weight on QoL (IWQoL-Lite) were administered. Spearman's rank correlation analysis was performed. A P value of less than 0.01 was used to set the statistical significance. A total of 117 patients (mean age: 47.4 years, mean BMI: 43.7) were enrolled. Correlations between WHO-DAS II and IWQoL-Lite were between 0.21 and 0.78. BMI between 0.19 and 0.26 correlated with WHO-DAS II and BMI between 0.23 and 0.49 correlated with IWQoL-Lite. In conclusion, low/moderate correlations between BMI index, disability, and health-related QoL measures, and a low association between the two outcome measures are reported, supporting the idea that they underline different and not transposable dimensions. PMID- 21629125 TI - Walk ratio (step length/cadence) as a summary index of neuromotor control of gait: application to multiple sclerosis. AB - In healthy adults, the step length/cadence ratio [walk ratio (WR) in mm/(steps/min) and normalized for height] is known to be constant around 6.5 mm/(step/min). It is a speed-independent index of the overall neuromotor gait control, in as much as it reflects energy expenditure, balance, between-step variability, and attentional demand. The speed independence of the WR in patients with multiple sclerosis (MS), and its capacity to discriminate (a) across patients with MS and controls and (b) among disability levels in MS were tested. The WR was computed in 30 outpatients with MS [20 women, 10 men; Extended Disability Status Scale (potential range: 0-10, observed median 3.5, range 2.5 5.0)] walking at free speed (range: 0.43-1.67 ms), and in 30 healthy controls (20 women, 10 men) at free and slow speed (range: 0.55-1.67 ms). The WR was 6.38+/ 0.66 in controls versus 5.36+/-0.86 in patients with MS (P<0.000), independent of age, sex, and walking speed. The WR was 5.95+/-0.69 and 4.90+/-0.70 in patients with an Extended Disability Status Scale score (P<0.001) below or above the median, respectively, independent of the disease duration (P<0.000). In patients with MS, the WR is a disability-sensitive index of neuromotor control of gait, and thus a promising outcome measure for treatments aimed at improving motor coordination. PMID- 21629126 TI - Corticosteroid exposure not associated with long-term bone mineral density in pediatric liver transplantation. AB - OBJECTIVES: The aim of the study was to examine the association of corticosteroid exposure and other skeletal risk factors with bone mineral density (BMD) and fractures following pediatric liver transplantation (LT) at a large single center. PATIENTS AND METHODS: Lumbar spine BMD, measured using dual-energy x-ray absorptiometry (DXA), was corrected for bone age in 52 ambulatory children ages 4 to 18 years, at least 1 year post-LT. Potential risk factors for skeletal health such as corticosteroid exposure, dietary and lifestyle factors, and growth and fracture occurrence, were related to BMD using univariate and multivariate regression analyses. RESULTS: The prevalence of low BMD (z score <-2) and post-LT fractures was 3 of 52 (5.8%) and 11 of 52 (21%), respectively. Univariate analysis revealed age >10 years at LT and body mass index (BMI) < 85th percentile at time of DXA were significantly associated with BMD (both P = 0.02). BMD did not correlate with corticosteroid dosage in the first year post-LT, the year before DXA or cumulative lifetime exposure. A cholestatic primary LT indication, acute rejection episodes, and fractures post-LT were not associated with BMD. Extracurricular physical activity, vitamin D, and calcium intake were not associated with BMD or fractures. Multivariate linear regression revealed increased time post-LT (P = 0.04) and higher BMI z score at time of DXA (P = 0.02) as the strongest independent variables associated with greater BMD. CONCLUSIONS: Neither corticosteroid exposure nor a cholestatic primary indication for LT influenced BMD, which was largely normal in this ambulatory group. Children and adolescents undergoing LT after the age of 10 years and those with low BMI post-LT may be at greatest risk of poor skeletal health later in life, and thus a potential target patient population to benefit from preventive interventions. PMID- 21629128 TI - Autoimmune hepatitis type 2 in a child with IPEX syndrome. PMID- 21629129 TI - Simultaneous distal radial fractures and carpal bones injuries in children: a review article. AB - Accurate diagnosis is crucial in effectively managing and treating both the radial and carpal fractures. Injuries to the carpal bones are not usually associated with fractures of the distal radius; however, the presence of a distal radius fracture does not preclude an associated carpal injury. The purpose of this review article is to compile cases of past reported distal radius fractures and simultaneous carpal injuries to restate its low prevalence within the pediatric population while still serving as a collective reference for management and treatment. After an electronic literature search was performed, we identified 18 published articles and 28 cases dealing with combined distal radial and carpal bones injuries in the pediatric population. As the mechanism of injury is similar for both fractures, despite the low incidence, orthopedic surgeons need to rule out carpal injury with a distal forearm fracture. Failure to treat both injuries appropriately may result in an unsatisfactory clinical result. PMID- 21629127 TI - Mechanisms of lipotoxicity in NAFLD and clinical implications. AB - With the epidemic of childhood obesity, nonalcoholic fatty liver disease (NAFLD) has become the most common cause of chronic liver disease in pediatrics. NAFLD is strongly associated with insulin resistance and increased level of serum free fatty acids (FFAs). FFAs have direct hepatotoxicity through the induction of an endoplasmic reticulum stress response and subsequently activation of the mitochondrial pathway of cell death. FFAs may also result in lysosomal dysfunction and alter death receptor gene expression. Lipoapoptosis is a key pathogenic process in NAFLD, and correlates with progressive inflammation, and fibrosis. Accumulation of triglyceride in the liver results from uptake and esterification of FFAs by the hepatocyte, and is less likely to be hepatotoxic per se. To date, there are no proven effective therapies that halt NAFLD progression or unfortunately improve prognosis in children. The cellular mechanisms of lipotoxicity are complex but provide potential therapeutic targets for NAFLD. In this review we discuss several potential therapeutic opportunities in detail including inhibition of apoptosis, c-Jun-N-terminal kinase, and endoplasmic reticulum stress pathways. PMID- 21629130 TI - Variable cardiac 18FDG patterns seen in oncologic positron emission tomography computed tomography: importance for differentiating normal physiology from cardiac and paracardiac disease. AB - PURPOSE: Cardiac fluorine-18-fluorodeoxyglucose (FDG) uptake is known to be variable in fasting oncologic positron emission tomography computed tomography studies. Increased posterolateral and basal FDG activity have been reported with the basal pattern ascribed to radiation injury. The purpose of this study was to investigate the spectrum of normal cardiac FDG findings seen in oncologic patients. MATERIALS AND METHODS: Men <35 years of age and women <45 years of age seen over a 3-year period were included. A visual assessment of FDG cardiac activity was performed using a 12-segment model of the heart by 2 observers. Focal papillary muscle activity was not included in the analysis. RESULTS: Of 65 patients who met the entry criteria, increased FDG activity was observed in the base of the heart in 37 (57%) patients. This was most common in the lateral-basal wall in 35 (54%) patients, followed by posterior-basal wall in 21 (32%) patients, anterior-basal wall in 10 (15%) patients, and basal-septum in 10 (15%) patients. Suppression of total cardiac activity was present in only 6 (9%) patients in spite of adequate fasting. Diffuse cardiac activity was seen in 9 (14%) patients. The previously reported increased posterolateral pattern was present in only 9 (14%) patients. CONCLUSION: This study confirms variable fasting FDG cardiac activity with a predominant basal pattern not associated with radiation injury. Knowledge of these patterns is important for recognition of possible underlying cardiac ischemia, tumor, or other inflammatory conditions encountered during interpretation of oncologic positron emission tomography computed tomography studies. PMID- 21629132 TI - Establishment of a primary pleomorphic xanthoastrocytoma cell line: in vitro responsiveness to some chemotherapeutics. AB - BACKGROUND: Anaplastic pleomorphic xanthoastrocytoma is an aggressively growing, malignant, and eventually fatal tumor of the central nervous system. Testing chemotherapeutic drug sensitivity under in vitro conditions would be a useful strategy to determine sensitive or resistant drugs for fatal brain cancers. OBJECTIVE: To establish primary cell cultures of excised tumor tissue from pleomorphic xanthoastrocytoma-bearing patients and to test their sensitivity against various anticancer chemotherapy drugs. METHODS: Prepared suspensions of the excised tumor tissue from a patient who had a recurrent grade 3 pleomorphic xanthoastrocytoma was cultured in culture dishes until cells began to grow. Immunofluorescent and immunohistochemical visualizations were performed using confocal and light microscopy. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide) assay in comparison with 3H-thymidine incorporation assay was used to test cellular toxicity of several anticancer drugs. RESULTS: We established vigorously growing primary cells of the tumor. Drug sensitivity testing was conducted successfully. CONCLUSION: Primary cell cultures of surgically removed tumor tissues may be useful in studies of cancer biology and chemotherapeutic drug sensitivity for recurrent malignant brain tumors, particularly for anaplastic pleomorphic xanthoastrocytoma. PMID- 21629133 TI - Strategy for treating unruptured vertebral artery dissecting aneurysms. AB - BACKGROUND: The natural course of unruptured vertebral artery dissecting aneurysms (VADAs) remains unclear. OBJECTIVE: The purpose of this retrospective study was to develop a strategy for treating unruptured VADAs based on long-term follow-up. METHODS: Our study population consisted of 100 patients with unruptured VADAs; in 66, the initial symptom was headache only, 30 presented with ischemic symptoms and 4 with mass effect. All underwent magnetic resonance imaging and magnetic resonance angiography at the time of admission and 2 weeks and 1, 3, 6, 12, and 24 months after the onset. If the dissection site was demonstrated to be enlarged on magnetic resonance imaging and magnetic resonance angiography without the manifestation of new symptoms, the patients received additional treatment to prevent bleeding. RESULTS: Of the 100 patients, 4 underwent early intervention because of symptom exacerbation. The other 96 were initially treated conservatively; during follow-up, 5 manifested lesion enlargement on magnetic resonance angiography. Nine patients received additional treatment; 1 underwent direct surgery with trapping of the dissection site, and 8 underwent coil embolization. The other 91 patients continued to be treated conservatively; the dissection site remained unchanged in 70, improved or healed in 18, and disappeared in 3 patients. We treated 38 patients with recurrent ischemic attacks with antiplatelet therapy. No patients experienced bleeding or permanent neurological deficits during follow-up. CONCLUSION: The nature of an unruptured VADA is not highly aggressive. However, if the dissection site enlarges without the manifestation of new symptoms, it should be occluded. In patients with recurrent ischemic attacks antiplatelet therapy should be considered. PMID- 21629134 TI - Ocular manifestations of xeroderma pigmentosum at a tertiary eye care center in Saudi Arabia. AB - PURPOSE: To study the clinical profile of Saudi cases of xeroderma pigmentosum (XP). DESIGN: This is a single-center, retrospective, consecutive case series of all cases of XP seen at King Khaled Eye Specialist Hospital from January 1, 1986, to December 31, 2006. The main outcome measures were clinical features, visual outcome, and histopathologic findings. RESULTS: Of 33 patients initially included in this study, 6 were excluded from the final analysis because of either unconfirmed diagnosis of XP or insufficient follow up. The final analysis included 27 patients (14 female patients, 13 male patients). A history of consanguinity was present in the parents of one third of our patients. The age at onset of ocular complications ranged from 5 to 67 years (median age, 19 years). The number of patients with no light perception (NLP) increased from 1 patient (3.7%) at the initial visit to 5 patients (18.5%) at the last visit. In 13 of 27 patients (48.1%), conjunctival tumors were observed; 10 tumors were confirmed histopathologically to be squamous cell carcinomas. Basal cell carcinoma (BCC) was the most common histopathologic diagnosis of eyelid tumors and was noted in 4 patients (14.8%). Furthermore, 10 patients (37.0%) had a history of skin malignancy, and 5 patients (18.1%) had neurologic abnormalities. CONCLUSION: Patients with XP who presented to a tertiary eye care center in Saudi Arabia had a high percentage of consanguinity in parents and late onset of ocular complications. Additionally, XP behaved aggressively in both malignancy profile and visual outcome. PMID- 21629135 TI - Direct fixation of extraocular muscles to a silicone sphere: a cost-sensitive, low-risk enucleation procedure. AB - PURPOSE: To describe a simple, cost-sensitive enucleation technique and to evaluate it in the context of intended benefits and long-term outcomes of alternate methods. METHODS: A retrospective record review of patients who underwent enucleation using the described technique at a single institution by 2 surgeons. Surgery involved direct fixation of extraocular muscles to a solid silicone sphere, using nonabsorbable braided sutures with knots tied beneath the muscle insertions. Outcome measures included implant-related complications. RESULTS: Seventy-five patients meeting inclusion criteria ranged in age from 3 to 94 years (mean, 54 years; median, 56 years). Surgical indications included a blind painful or disrupted eye in 56 cases; uveal melanoma in 15 cases; retinoblastoma in 2 cases; and endophthalmitis in 2 cases. Follow-up intervals ranged from 3 to 57 months (mean, 27 months; median, 22 months). Complications requiring surgical revision of the implant occurred in 2 of the 75 cases (one luxation; one exposure). There were no instances of chronic socket discharge or implant infection. CONCLUSIONS: Nonabsorbable-suture attachment of muscles to a solid silicone implant offers an inexpensive enucleation option, with minimal risk of implant migration, exposure, or infection. In light of widespread disaffection with pegging of porous implants, and with no motility advantage of unpegged porous over nonporous implants, consideration should be given to techniques that are equally effective, less costly, and perhaps more reliable. PMID- 21629136 TI - Orbital cellulitis with subperiosteal abscess: demographics and management outcomes. AB - PURPOSE: Infectious orbital cellulitis represents a serious threat to vision and, if untreated, poses significant morbidity risk. In this study, the authors attempt to further characterize the features of orbital cellulitis with subperiosteal abscess (SPA) and determine outcomes based on the type of surgical intervention employed. METHODS: Data were obtained by retrospective chart review of all inpatient admissions for orbital inflammation/cellulitis from Sept 2005 to April 2010. Charts were reviewed for demographic information, radiographic and clinic evidence of orbital cellulitis, presence of SPA (defined by radiographic criteria), interventions taken (surgical and nonsurgical), presence of concurrent sinusitis, types of microbes present, and duration of hospital admission. Statistical analysis was performed using chi-square tests. RESULTS: A total of 97 patients were admitted with a diagnosis of orbital inflammation/cellulitis, of whom 49 patients had clinical and/or radiographic evidence of orbital cellulitis. Twenty-four patients had radiographic evidence of SPA. All patients with SPA had concurrent sinusitis, and 16 patients underwent initial surgical intervention. Nine patients had external transcaruncular/transcutaneous SPA drainage only, 6 had combined SPA and sinus drainage, and one had sinus drainage alone. Of those that underwent SPA-only drainage, 5 had SPA reaccumulation, while no reaccumulation occurred with combined SPA and sinus drainage. No reaccumulation occurred if the SPA was less than 2 cm in its greatest diameter. CONCLUSION: In this study, for those abscesses larger than 2 cm, combined sinus and SPA drainage was associated with improved treatment outcome, demonstrated by absence of abscess reaccumulation and shorter hospital stay. SPA-only drainage was more frequently associated with SPA reaccumulation. PMID- 21629137 TI - Diagnosis and management of an intraorbital vascular fistula. AB - A spontaneous intraorbital arteriovenous fistula is an extremely rare phenomenon. More commonly, arteriovenous fistulas occur spontaneously as carotid cavernous fistulas and dural cavernous fistulas or are secondary to trauma. Surgical approaches vary widely and each carries their own set of potential problems. The purpose of this single case report was to provide an example of a successful treatment of a rare, spontaneous intraorbital vascular fistula. PMID- 21629138 TI - Primary adenoid cystic carcinoma: an extremely rare eyelid tumor. AB - Cutaneous adenoid cystic carcinoma is an extremely rare clinical entity. Among the few cases reported in the literature, most had contiguous involvement from the lacrimal gland. Primary adenoid cystic carcinoma is one of the rarest eyelid tumors. The authors report a case of adenoid cystic carcinoma arising from the lower eyelid. PMID- 21629139 TI - The effect of tissue wrapping on the expansion of hydrophilic orbital implants. AB - PURPOSE: Hydrogel implants have been used in hypoplastic orbital expansion. Advantages include insertion through a small incision and expansion without manipulation. The rapid rate of expansion, however, may lead to undesirable outcomes, including implant migration. The authors evaluate the effect of tissue wrapping on expansion of hydrogel implants. METHODS: Three identical hydrogel implants were subjected to one of 3 in vitro interventions: unwrapped control, porcine scleral-wrapped, and porcine fascia-wrapped. Each implant was submerged in normal saline solution. Diameter and volume were measured hourly for 24 hours and then every 4 hours until a plateau or maximal weight was achieved. Measurements were plotted with time as the independent variable and the above measurements as dependent variables. In addition, the best exponential function was graphed and the area under the curve was calculated. RESULTS: Linear plots of data showed a reduction in the rate of diameter and volume expansion for wrapped implants. The areas under the curve of the best-fit exponential function were less for both scleral- and fascia-wrapped implants. Fascia wrapping slowed the rate of volume expansion more than sclera. Also, fascia wrapping seemed to significantly reduce the final implant diameter and volume. CONCLUSIONS: Tissue wrapping slows the rate of expansion of hydrogel spheres in vitro. Fascia wrapping may also reduce the final implant size. PMID- 21629140 TI - Lateral canthoplasty with acellular cadaveric dermal matrix graft (AlloDerm) reinforcement. AB - Lateral canthal dystopia can lead to lower eyelid malposition, abnormal lateral eyelid fissure appearance, and lagophthalmos. In most cases, the lateral canthus can be repaired with a standard lateral canthopexy or canthoplasty. In a fraction of cases, especially when recurrent, the surgical repair may require additional lateral canthal reinforcement. PMID- 21629141 TI - Management of perineural invasion in sebaceous carcinoma of the eyelid. AB - PURPOSE: To report the occurrence and management of perineural invasion in patients with sebaceous carcinoma of the eyelid. METHODS: An ophthalmology database was searched for all patients treated for sebaceous carcinoma of the eyelid by the principal investigator between May 1999 and May 2010. The clinical records and pathology specimens of the patients with microscopic perineural invasion as an incidental finding in their eyelid surgical specimen were reviewed. RESULTS: Forty-two patients with sebaceous carcinoma of the eyelid were treated by the principal investigator during the study period. Three of them had evidence of microscopic perineural invasion in the surgical specimen. Each patient was treated with surgery with or without radiotherapy. The first patient underwent orbital exenteration and negative sentinel lymph node biopsy, subsequently developed distant metastasis, and died 20 months after exenteration. The second patient underwent resection of the tumor and positive sentinel lymph node biopsy, postoperative irradiation of the eyelid, completion neck dissection and parotidectomy for the positive sentinel lymph node, and irradiation of nodal basins; she was free of disease at last follow up (12 months after tumor resection). The third patient underwent resection of the tumor, developed regional lymph node metastasis 3 months later, underwent lymph node dissection and postoperative nodal irradiation, and was free of disease at last follow up (9 months after tumor resection). CONCLUSIONS: Perineural invasion was encountered in 7% of patients with eyelid sebaceous carcinoma. The authors' preference is to treat patients with an incidental finding of microscopic perineural invasion with postoperative adjuvant radiotherapy, ideally within 4 to 6 weeks after surgical resection of the primary eyelid tumor. PMID- 21629142 TI - Novel management of the microphthalmic orbit in a patient with PHACE syndrome. AB - A full-term female infant with severe unilateral microphthalmos and ipsilateral orbital hemangioma in the setting of PHACE syndrome was successfully managed with oral propranolol therapy and subsequent orbital conformer placement. After 3 months of systemic propranolol therapy, the orbital hemangioma had shown significant regression without systemic or local complications. The authors believe this to be the first reported use of systemic propranolol therapy in the management of a hemangioma within a microphthalmic orbit of a patient with PHACE syndrome. PMID- 21629143 TI - Pediatric alternate site of care during the 2009 H1N1 pandemic. AB - OBJECTIVES: This study aimed to describe the design and implementation of an alternate site of care (ASC) for nonurgent pediatric patients with influenza-like illnesses during the 2009 H1N1 pandemic and to evaluate its performance. METHODS: We describe the design and physical implementation of an ASC. Evaluation of the utilization, patient demographics, throughput, safety, family satisfaction, and cost are presented. RESULTS: The process of project development, site selection, clinical algorithms, staffing supplies, and cost are detailed. The ASC was used for 7.5 days, and 137 patients were treated. The median age was 6.5 years. Forty five percent were male, and English was the primary language. Median length of stay for patients evaluated was 65 minutes. Of patients, 5.8% were transferred from the ASC to the ED for further care. Also, 2.3% of patients returned to the ED within 72 hours; however, none required admission. There were no adverse events associated with the ASC and 92% of families rated overall care as very good or excellent. CONCLUSIONS: Selected nonurgent patients with influenza-like illness during a pandemic can be treated in a safe and timely manner with high levels of family satisfaction in a novel setting. PMID- 21629144 TI - A content analysis of parents' written communication of needs and expectations for emergency care of their children. AB - OBJECTIVE: We investigated the potential value of information shared by parents on a written form designed to capture needs and expectations for care to an emergency department (ED) system that values patient-centeredness. METHODS: We conducted a retrospective content analysis of parent-completed written forms collected during an improvement project focused on parent-provider communication in a pediatric ED. The primary outcome was potential value of the completed forms to a patient-centered ED system, defined as a form that was legible, included observations that mapped to medical problems, and included reasonable parental requests. We analyzed variation in potential value and other form attributes across a priori-defined visit type and acuity. Visit type was validated by a separate, blinded medical record review. RESULTS: A random stratified sample of 1008 forms was established from 6937 parent-completed forms collected during the 6-month improvement project; 995 of 1008 forms had matching medical records; 922 (92.7%) of 995 forms demonstrated potential value; 990 (99.5%) of 995 forms were legible; 948 (95.3%) of 995 forms included observations that mapped to a medical problem, and 599 (93.3%) of 642 forms contained reasonable parental requests. There was good agreement between the form and medical record for visit type (kappa = 0.62). The potential value of forms did not vary significantly across visit type (88.2%-92.8%) or acuity (88.9%-93.4%). CONCLUSIONS: Information shared by parents on written forms designed to capture needs and expectations provides potential value to a patient-centered ED system. The high level of informational value is consistent across patient type and acuity level. PMID- 21629145 TI - Assessing survey methods and firearm exposure among adolescent emergency department patients. AB - OBJECTIVE: The objective of the study was to determine the nature of gun exposure in an adolescent population presenting to an urban emergency department (ED) and to ascertain attributes that correlate with the ability to buy a gun or to access a loaded gun within 3 hours. METHODS: A convenience sample of adolescents aged 12 to 17 years presenting to a single ED was surveyed from September 2005 to February 2006. The survey tool was derived from the Centers for Disease Control and Prevention Youth Risk Behavior Surveillance Study and the Hamilton Youth and Guns Poll with additional questions related to gun accessibility. RESULTS: Of the 300 total participants, 28% could access a loaded gun in less than 3 hours, and 22% stated that they could easily buy a gun. A significant increase in the ability to buy a gun in ninth grade (27%) compared with eighth grade (6%) was found. Independent predictors of the ability to buy a gun include gang membership, drug use, male sex, and witnessing a gun at school. Independent predictors of being able to access a loaded gun within 3 hours include having a gun in the home and witnessing a gun at school. CONCLUSIONS: The ability to buy a gun and access to a loaded gun within 3 hours are relatively common among ED adolescent patients. Having a gun in the home and witnessing a gun at school were independent predictors of the ability to access a loaded gun within 3 hours. Gang membership and drug use were associated with a self-reported ability to buy a gun. PMID- 21629146 TI - Is there a role for intravenous acetaminophen in pediatric emergency departments? AB - BACKGROUND: As a nonopioid parenteral analgesic intravenous (IV) acetaminophen is potentially attractive for emergency department (ED) use. However, there is little experience with its use in the pediatric ED setting. We introduced the agent into a pediatric ED with a preliminary restrictive prescribing regimen and describe its use. METHODS: This is a retrospective record review of all patients who had received IV acetaminophen over 12 months. Prescribing indications were for analgesia only (not for fever management) in patients at risk of opioid related adverse events. We assessed the demographics, dosing, presenting complaints, discharge diagnoses, and indications for IV acetaminophen use. RESULTS: Thirty-one patients received IV acetaminophen (mean age, 10 years). All patients were at least moderately sick according to their triage assessment. The median dose was 15 mg/kg with a median of 1 dose administered. Presenting complaints were mainly trauma, abdominal complaints, and sepsis/fever/neutropenia. Thirty-nine percent of patients had severe underlying conditions. Two patients died: 1 patient had a chronic neurological condition, and the other was undergoing palliative treatment for cancer. Physicians complied with prescribing indications for half the patients: 53% were at risk of opioid related adverse events. Physicians prescribed outside the initial indications as part of multimodal analgesia (13%), for complex patients with fever and pain (7%), and for fever only in patients unable to tolerate enteral administration (27%). CONCLUSIONS: In the setting of an initial restrictive prescribing regimen, IV acetaminophen was used in a small number of pediatric ED patients. In addition to use in patients at risk of opioid adverse events, the medication was also used in complex patients who were unable to tolerate an enteral formulation. Emergency department prescribing guidelines have been modified accordingly. PMID- 21629147 TI - Illicit drug exposure in patients evaluated for alleged child abuse and neglect. AB - BACKGROUND: Substantiation of drug exposure in cases with alleged maltreatment is important to provide proper treatment and services to these children and their families. A study performed at University of Iowa Hospitals and Clinics showed that 30% of pediatric patients with burn injuries, which were due to child maltreatment, were also exposed to illicit drugs. OBJECTIVE: The children presenting to the University of Iowa Hospitals and Clinics with alleged maltreatment have been tested for illicit substances since 2004. The objective of this study was to analyze the presence of illicit drug exposure in the pediatric subpopulation admitted to pediatric inpatient and outpatient units for an evaluation for abuse/neglect. DESIGN AND METHODS: The study design is a retrospective chart review. Using hospital databases, every pediatric chart with a child abuse/neglect allegation was retrieved. The association between risk factors and clinical presentation and illicit drug test result was assessed. Excel and SAS were used for statistical analysis. Institutional review board approval was obtained to conduct this study. RESULTS: Six hundred sixty-five charts met study inclusion criteria for child abuse/neglect allegation. Of those, 232 cases were tested for illicit drugs between 2004 and 2008 per the testing protocol. Thirty-four cases (14.7%) tested positive on a drug test. Positive test rates based on clinical presentation were 28.6% (18/63) in neglect cases, 16.1% (5/31) in cases with soft tissue injuries, 14.3% (4/28) in burn injuries, 10.0% (2/20) in cases with sexual abuse, 7.1% (2/28) in cases with fractures, and 4.8% (3/62) in abusive head trauma cases. There were long-term abuse findings in 129 children (55.6%). Logistic regression analysis revealed that positive drug testing was most significantly associated with clinical symptoms suggesting physical abuse or neglect versus sexual abuse (odds ratio [OR] = 6.70; 95% confidence interval [CI], 1.26-35.49; P = 0.026), no or public health insurance versus those with private insurance (OR = 4.49; 95% CI, 1.47-13.66; P = 0.008), history of parental drug abuse versus those without parental history of drug abuse (OR = 3.42; 95% CI, 1.38-8.46; P = 0.008), and history of domestic violence versus those without a history of domestic violence (OR = 2.81; 95% CI, 1.08 7.30; P = 0.034). CONCLUSIONS: The results of this study showed that an illicit drug screening protocol used in the assessment of children evaluated for child abuse identified almost 15% of the population of allegedly abused and neglected children who were tested according to a protocol being exposed to illicit drugs. Thus, routine drug testing of at least children assessed for neglect and nonaccidental burn and soft tissue injuries, children with a history of either parental drug use or domestic violence is recommended. PMID- 21629148 TI - Predictors of psychiatric boarding in the pediatric emergency department: implications for emergency care. AB - OBJECTIVES: Patients who present to the emergency department (ED) and require psychiatric hospitalization may wait in the ED or be admitted to a medical service because there are no available inpatient psychiatric beds. These patients are psychiatric "boarders." This study describes the extent of the boarder problem in a large, urban pediatric ED, compares characteristics of psychiatrically hospitalized patients with boarders, and compares predictors of boarding in 2 ED patient cohorts. METHODS: A retrospective cohort study was conducted in 2007-2008. The main outcome measure was placement into a psychiatric facility or boarding. Predictors of boarding in the present analysis were compared with predictors from a similar study conducted in the same ED in 1999 2000. RESULTS: Of 461 ED patient encounters requiring psychiatric admission, 157 (34.1%) boarded. Mean and median boarding duration for the sample were 22.7(SD, 8.08) and 21.18 hours, respectively. Univariate generalized estimating equations demonstrated increased boarding odds for patients carrying Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition diagnoses of autism, mental retardation, and/or developmental delay (P = 0.01), presenting during the weekend (P = 0.03) or presenting during months without school vacation (P = 0.02). Suicidal ideation (SI) significantly predicted boarding status, with increased likelihood of boarding for severe SI (P = 0.02). Age, race, insurance status, and homicidal ideation did not significantly predict boarding in the 2007 2008 patient cohort, although they did in the earlier study. Systemic factors and SI predicted boarding status in both cohorts. CONCLUSIONS: Suicidal patients continue to board. Limits within the system, including timing of ED presentation and a dearth of specialized services, still exist, elevating the risk of boarding for some populations. Implications for pediatric ED psychiatric care delivery are discussed. PMID- 21629149 TI - Is the Broselow tape a reliable indicator for use in all pediatric trauma patients?: A look at a rural trauma center. AB - OBJECTIVE: The purpose of this study was to determine the effectiveness of the Broselow tape in the evaluation of pediatric trauma patients. METHODS: The trauma registry of a rural level I trauma center was examined. All pediatric trauma patients 16 years or younger were reviewed from 2002 to 2006, totaling 2358 patients. The Broselow tape measures to 146.5 cm. Patients whose height correlated with the tape and had their heights and weights in the medical record were included. The constant variable was the heights by which the estimated weights of the Broselow tape were compared with the actual weights of the patients. RESULTS: A total of 657 patients matched this height and had both heights and weights in their record. Most children (349/657; 53.1%) fell outside the predicted weight range, and of these, 77.1% of the actual weights were greater than those predicted by the Broselow scale. This is observed across all age groups. In patients with heights less than 75 cm, two thirds of patients' weights correlated with the Broselow estimated weight; however, those that deviated did so by 2 to 3 color intervals larger. This deviation was statistically significant in all groups. CONCLUSIONS: In our population, the Broselow tape is an ineffective tool to predict weight in more than 50% of pediatric trauma patients. This may lead to the underdosing of emergency medications and blood products. PMID- 21629150 TI - Interexaminer agreement in physical examination for children with suspected soft tissue abscesses. AB - OBJECTIVE: This study aimed to measure interexaminer agreement for physical examination (PE) findings in children with a suspected soft tissue abscess. METHODS: A prospective study was conducted from March 1 to July 31, 2007, at an urban, tertiary care children's hospital emergency department. Children presenting to the emergency department with a suspected local skin abscess were independently examined by 2 physicians. Interrater agreement of 7 PE findings for children with a suspected soft tissue abscess was assessed. Interrater agreement was calculated for the diagnosis of the lesion and decision to incise and drain. RESULTS: A total of 105 paired observations were completed by a total of 27 physicians. The patients examined were aged 2 weeks to 18 years, with a mean age of 80 months. Lesions were most frequently encountered on the buttocks (38%). Incision and drainage was attempted in 75% of cases, with purulent material obtained in 92% of all attempts. Interrater agreement was substantial for erythema (kappa = 0.66) and size of the lesion (intraclass correlation coefficient = 0.78), moderate for drainage (kappa = 0.57) and tenderness (kappa = 0.40), fair for fluctuance (kappa = 0.35), and poor for warmth (kappa = 0.15) and showed no agreement for induration (kappa = -0.08). There was moderate agreement on diagnosing the lesion as an abscess (kappa = 0.48) and determination if the lesion required incision and drainage (kappa = 0.44). CONCLUSIONS: Interexaminer agreement of examination findings and diagnosis of an abscess was fair to moderate, implying a lack of precision of PE as the primary means for diagnosis. Future studies of diagnostic adjuncts, such as bedside ultrasonography, may lead to improved management of soft tissue infections in children. PMID- 21629151 TI - "Inappropriate" pediatric emergency medical services utilization redefined. AB - OBJECTIVES: Previous reports on emergency medical services (EMS) transportation of pediatric patients have demonstrated a high rate of overutilization. However, there is also a concern that pediatric patients may underutilize EMS for emergencies that might benefit from EMS. This article compares EMS utilization rate between adult and pediatric patients for high-acuity patients and for the most common reasons for transport. METHODS: This study was a secondary analysis of the National Hospital Ambulatory Medical Care Survey to compare hospital arrival by EMS to walk-in arrivals. Primary variables were age category, mode of arrival, immediacy to be seen (triage category), reason for visit, and disposition. RESULTS: There were 253,898 records, weighted to represent 914.4 million emergency department visits, included. Emergency medical services mode of arrival was significantly higher for adult patients at 19.1% as compared with pediatric patients at 6.5% (odds ratio, 3.38). For the subgroup of patients requiring critical care interventions, adult patient arrival by EMS was 87.3% as compared with pediatric patients at 66.3% (odds ratio, 3.50). When considering the top 20 most common medical complaints in which pediatric patients used EMS transport, adult patients utilized EMS more frequently in 85% (17/20) of those complaints. CONCLUSIONS: As compared with adults, pediatric patients are less likely to utilize EMS for transport to the hospital for both routine and emergent complaints. The definition of inappropriate utilization of EMS for pediatric transport, which has largely focused on inappropriate overutilization, should also incorporate the potential of underutilization for critical patients. PMID- 21629152 TI - Parental health literacy and asthma education delivery during a visit to a community-based pediatric emergency department: a pilot study. AB - OBJECTIVE: The objective of the study was to compare change in asthma knowledge among parents with low or adequate health literacy after video or written asthma education delivered during their child's asthma-related emergency department (ED) visit. METHODS: We recruited a convenience sample of parents presenting to the ED with their asthmatic child 2 to 14 years old. Parents were randomized to receive video (intervention) or written (active-control) asthma education materials. Health literacy levels were determined using the Rapid Estimate of Adult Literacy in Medicine tool. Asthma knowledge was evaluated at enrollment and 6-week telephone follow-up. Differences in knowledge scores were analyzed using Wilcoxon signed rank tests; chi2 tests were used for comparisons of categorical variables. RESULTS: Of 150 eligible parents, 129 participated (86%), and 117 were eligible for follow-up. Telephone follow-up was completed with 86 parents (74%). Health literacy levels were low in 31% of the parents. High asthma knowledge scores at enrollment were achieved by 33% of low-literacy and 59% of adequate-health literacy parents (P = 0.025). Improvement in knowledge was realized for low literacy parents regardless of the type of education (P < 0.001). Parents with adequate health literacy showed increased knowledge scores only after viewing the video. CONCLUSIONS: Asthma education materials distributed at the time of an ED visit increase parental knowledge about the disease. Video-based asthma education appears promising as a tool for increasing asthma knowledge in both low- and adequate-health-literacy parents. PMID- 21629153 TI - Myocardial response in preterm fetal sheep exposed to systemic endotoxinaemia. AB - Exposure of the fetus to antenatal inflammation can occur from chorioamnionitis, which may progress to a fetal inflammatory response syndrome (FIRS) and to fetal sepsis. We tested whether the fetal myocardium responded to systemic Gram negative endotoxinaemia. We hypothesized that the myocardium would respond to inflammation by changes in hypoxia-inducible factor-alpha (HIF-1alpha), inducible NO-synthase (iNOS), Toll-like receptors 2 and 4 (TLR2 and TLR4), IL-6, and phosphorylated signal transducer and activator of transcription-3 (pSTAT3). To model systemic endotoxinaemia, fetal sheep were exposed to Gram-negative endotoxin or saline i.v. 3 d before preterm delivery at 113 d of gestation (term = 147 d). All endotoxin-exposed animals developed cardiac dysfunction within these 72 h. Cardiac mRNA and protein levels of HIF-1alpha and TLR2 and TLR4 mRNA increased, whereas STAT3 phosphorylation decreased significantly. IL-6 and iNOS mRNA remained unchanged. Fetal systemic endotoxinaemia induced myocardial inflammation by activating TLR2 and 4. The following cardiac dysfunction seems not to be mediated via cardiac iNOS. PMID- 21629154 TI - The influence of maternal body mass index on infant adiposity and hepatic lipid content. AB - Maternal overweight and obesity are associated with adverse offspring outcome in later life. The causal biological effectors are uncertain. Postulating that initiating events may be alterations to infant body composition established in utero, we tested the hypothesis that neonatal adipose tissue (AT) content and distribution and liver lipid are influenced by maternal BMI. We studied 105 healthy mother-neonate pairs. We assessed infant AT compartments by whole body MR imaging and intrahepatocellular lipid content by H MR spectroscopy. Maternal BMI ranged from 16.7 to 36.0. With each unit increase in maternal BMI, having adjusted for infant sex and weight, there was an increase in infant total (8 mL; 95% CI, 0.09-14.0; p = 0.03), abdominal (2 mL; 95% CI, 0.7-4.0; p = 0.005), and nonabdominal (5 mL; 95% CI, 0.09-11.0; p = 0.054) AT, and having adjusted for infant sex and postnatal age, an increase of 8.6% (95% CI, 1.1-16.8; p = 0.03) in intrahepatocellular lipid. Infant abdominal AT and liver lipid increase with increasing maternal BMI across the normal range. These effects may be the initiating determinants of a life-long trajectory leading to adverse metabolic health. PMID- 21629155 TI - Longitudinal study of childhood adiposity and the risk of developing components of metabolic syndrome-the Da Qing children cohort study. AB - Childhood adiposity is increasingly recognized as a significant predictor of cardiometabolic risks in later life. The aim of this study was to investigate factors associated with longitudinal changes in weight during childhood and the development of metabolic disease risk factors. Four hundred twenty-four children from DaQing city, China, were recruited at 5 y old and followed up for 5 y. Birth weight, television (TV) viewing time at 5 y old, blood pressure, anthropometric measurements, fasting plasma insulin (FI), and triglycerides (TG) levels were measured at 5 and 10 y old. Both birth weight and TV viewing time at 5 y old significantly correlated with percentage of ideal weight for height (WFH) at 5 y old (WFH5; p = 0.0032 and p = 0.01), but only TV time was significantly correlated with WFH at 10 y old (WFH10; p < 0.0001). Blood pressures, FI, homeostasis model assessment for insulin resistance (HOMA-IR), and TG at 10 y old were significantly greater in those children who had greater change in WFH from 5 to 10 y old (DeltaWFH). We concluded that TV viewing time was the stronger determinant of later childhood adiposity. A greater DeltaWFH was associated with increased cardiometabolic risk factors at 10 y old. PMID- 21629156 TI - The pioneer gut microbiota in human neonates vaginally born at term-a pilot study. AB - The pioneer microbiota of the neonate may affect future actions of the immune system. This study aimed to map the pioneer microbiota in healthy neonates vaginally born at term. A subgroup of neonates born large for GA (LGA) was compared with the neonates appropriate for GA (AGA). Fecal samples were collected, within 48 h after birth, from 79 neonates. Quantitative PCR was used for enumeration of Lactobacillus, a subgroup of Lactobacillus common in the vagina, Bifidobacterium, Enterococcus, Enterobacteriaceae, and the Bacteroides fragilis group. Cloning and sequencing were applied for subgroups of neonates born LGA or AGA. Lactobacillus was detected in all neonates, whereas other bacterial groups were detected only in 14 to 30% of the subjects. The prevalence of Gram-negative Proteobacteria was higher in neonates born LGA, whereas Gram positive Firmicutes was more prevalent in neonates born AGA (p < 0.001). This study contributed to increased knowledge of the pioneer microbiota and indicates that neonates born LGA had significantly different microbiota compared with those born AGA. As the early microbiota can be important for maturation of the immune system, the outcome from this study may be relevant in the care of pregnant woman and newborns. PMID- 21629157 TI - Isolation and culture of fibroblasts, vascular smooth muscle, and endothelial cells from the fetal rat ductus arteriosus. AB - The ductus arteriosus (DA), a fetal arterial shunt vessel between the proximal descending aorta and the pulmonary artery, closes shortly after birth. Initial functional closure as a result of the DA's smooth muscle contraction is followed by definite anatomical closure. The latter involves several complex mechanisms like endothelial cushion formation and smooth muscle cell migration resulting in fibrosis and sealing of the vessel. These complex steps indicate highly specialized functions of the DA vascular smooth muscle cells (VSMCs), endothelial cells, and fibroblasts. Herein, we describe a new reproducible method for isolating VSMCs, endothelial cells, and fibroblasts of high viability from fetal rat DA using immunomagnetic cell sorting. Purity of the different cell cultures was assessed by immunohistochemistry and flow cytometry and ranged between 85 and 94%. The capability of the VSMCs to react to hypoxic stimuli was assessed by intracellular calcium and ATP measurements and by VEGF mRNA expression analysis. VSMCs respond to hypoxia with decreases in intracellular calcium concentrations and ATP levels, whereas VEGF mRNA expression increased 3.2-fold. The purified vessel-specific different cell types are suitable for subsequent gene expression profiling and functional studies and provide important tools for improving our understanding of the complex processes involved in the closure of the DA. PMID- 21629158 TI - Sterility of posterior elements of the spine in posterior correction surgery. AB - STUDY DESIGN: Prospective clinical series. OBJECTIVE: To investigate the sterility of the posterior elements of the spine during posterior correction surgery. SUMMARY OF BACKGROUND DATA: Bacterial growth in the operating field is considered one of the causes of surgical-site infection. METHODS: A total of 80 consecutive patients (mean age = 19.5 years) who underwent posterior correction surgery for spinal deformities were included in the study. During surgery, specimens for bacterial culture were obtained by swabs from (1) the skin after cleansing using povidone-iodine scrub solution but before draping, (2) laminae immediately after exposure, (3) laminae immediately after screw placement, (4) laminae immediately before wound closure, (5) a small piece of bone obtained from the spinous process immediately after exposure and placed on a sterile dish during surgery as a control, kept covered, and (6) a similar sample as (5), kept uncovered. The culture was conducted on both blood agar and Gifu anaerobic medium agar plates. RESULTS: No patient developed surgical-site infection. The culture was positive in specimen (S) 1 in 25 patients (31.3%), S2 in 20 (25%), S3 in 25 (31.3%), S4 in 26 (32.5%), S5 in 6 (7.5%), and S6 in 7 (8.8%). Bacterial species included Propionibacterium acnes in 15 specimens, Propionibacterium species in 9, and others in 10. Of the 25 patients with a positive culture in S1 and 55 patients without, 22 (88%) and 26 (47.3%), respectively, demonstrated a positive culture in specimens obtained during surgery. CONCLUSION: The culture was more frequently positive in specimens obtained immediately before wound closure than in those harvested immediately after exposure and isolated from the surgical field. The results suggest that bacterial contamination of the operating field may originate from the skin of the patient during surgery. PMID- 21629159 TI - Relationship between preoperative expectations, satisfaction, and functional outcomes in patients undergoing lumbar and cervical spine surgery: a multicenter study. AB - STUDY DESIGN: Analysis of prospectively collected multicenter data. OBJECTIVE: To explore the relationship between preoperative expectations and postoperative outcomes and satisfaction in lumbar and cervical spine surgery. SUMMARY OF BACKGROUND DATA: Back pain is one of the most common health problems, leading to the utilization of health care resources, work loss, and sick benefits. Patient expectations influence posttreatment outcomes, both surgical and nonsurgical. There is little research on the importance of preoperative expectations in spine surgery. Existing studies evaluate the technical aspects of interventions and functional outcomes but fail to take into account patient expectations. The authors hypothesized that expectations dramatically affect spine patient satisfaction independent of functional outcomes. METHODS: Prospectively collected patient-entered data from patients undergoing lumbar and cervical spine surgery from 2 study centers collected using a Web-based patient health survey system were analyzed. The study included patients who underwent operative intervention (decompression with or without fusion) with at least a 3-month period of follow up. Preoperative expectations were measured using the Musculoskeletal Outcomes Data Evaluation and Management System's (MODEMS) expectation survey. Postoperative satisfaction and fulfillment of expectations were measured using the MODEMS satisfaction survey. Postoperative functional outcomes were measured using the Oswestry Disability Index and 36-item short form health survey. Ordinal logistic regression multivariate modeling was used to examine predictors of postoperative satisfaction. Linear regression multivariate modeling was used to examine predictors of functional outcomes. RESULTS: Greater fulfillment of expectations led to higher postoperative satisfaction and was associated with better functional outcomes. Higher preoperative expectations led to decreased postsurgical satisfaction but were associated with improved functional outcomes. Higher postoperative satisfaction was associated with improved functional outcomes and vice versa. Type of surgery also influenced satisfaction and function, with cervical patients being less satisfied but having better functional outcomes than lumbar patients. CONCLUSION: This study showed that more than functional outcomes matter; preoperative expectations and fulfillment of expectations influence postoperative satisfaction in patients undergoing lumbar and cervical spine surgery. This underlines the importance of taking preoperative expectations into account to obtain an informed choice on the basis of the patient's preferences. PMID- 21629160 TI - Extradural nodular fasciitis arising in the spinal canal. AB - STUDY DESIGN: Case report. OBJECTIVE: To describe a patient with nodular fasciitis arising in the lumbar extradural space. SUMMARY OF BACKGROUND DATA: Nodular fasciitis is a benign proliferation of fibroblasts and myofibroblasts. It commonly occurs in the subcutaneous tissue of an upper extremity, trunk, head, and neck, but rarely arises in the spinal canal. METHODS: A 7-year-old boy experienced gradually increasing intense radiating pain from the bilateral buttocks to the lower extremities after a bruise on his lower back. Computed tomography and magnetic resonance imaging demonstrated a relatively circumscribed mass in the dorsal epidural space from the first lumbar vertebra (L1) to L2. The presumptive diagnosis based on the radiologic findings included aggressive neoplasm such as extraskeletal Ewing sarcoma/primitive neuroectodermal tumor or malignant lymphoma. RESULTS: The patient underwent L1-L2 laminectomy and resection of the tumor. Histologically, the tumor was mainly composed of a proliferation of spindle cells without atypia, positive for vimentin and smooth muscle actin, and myxoid areas with a loosely textured feathery pattern. These findings are the typical features of nodular fasciitis. Surgery relieved the patient's pain, with no evidence of recurrence at a recent 4-year follow-up. CONCLUSION: This report presents a very rare case of extradural nodular fasciitis arising in the lumbar spinal canal, which could have been misinterpreted as a malignant tumor such as extraskeletal Ewing sarcoma/primitive neuroectodermal tumor because of its rapid growth and absence of distinguishing radiologic features. A detailed histopathologic examination including immunohistochemistry is important for the correct diagnosis. PMID- 21629161 TI - Lumbar sagittal shape variation vis-a-vis sex during growth: a 3-year follow-up magnetic resonance imaging study in children from the general population. AB - STUDY DESIGN: A longitudinal descriptive magnetic resonance imaging study on the changes of the supine lumbar lordosis (SLL), supine sacral slope (SSS), and sagittal wedging of the vertebral body (VB) and intervertebral discs (IVD) in children from the general population. OBJECTIVE: To compare the shape variation during growth of the SLL, SSS, and sagittal wedging of the VB and IVD in boys and girls. SUMMARY OF BACKGROUND DATA: The normal shape variation of the VB, IVD, SLL, and SSS during growth vis-a-vis sex in children from the general population is poorly clarified in the literature as it is usually reported in relation to pathological conditions. METHODS: The SLL, SSS, and sagittal wedging of all lumbar (L1-S1) VBs and IVDs were measured twice from T2-weighted magnetic resonance images of 100 healthy children (51 boys and 49 girls), at mean age 12 to 13 years (t0) and after approximately 3 years (t1) using the iQ-VIEW system (IMAGE Information Systems Ltd., Plauen, Germany). Data for body compositions and pubertal status were collected, and their correlations were analyzed. RESULTS: At baseline (t0), most lumbar VBs were significantly more lordotic in boys (17.1o) than in girls (22.2o); however, girls manifested greater SLL and SSS (45.2o and 33.6o, respectively) than boys (40.7o and 31.4o, respectively), and all IVDs were lordotic, with only the L5-S1 IVD differing between sexes, being more lordotic in girls than in boys (mean difference = 2.8o). At follow-up (t1), SSS became greater in boys (35.7o) than in girls (32.5o), yet all other parameters became independent of sex including all IVDs (except L5-S1) becoming significantly more lordotic, and more so in boys than in girls (total lumbar mean differences being 9.0o and 3.8o, respectively). Increase in boys' heights was correlated with the increase in the L2-L4 lordotic IVD wedging (r = 0.45). Positive correlations were indicated between puberty Tanner stage and individual's height and weight (0.41 < r < 0.45). CONCLUSION: Lumbar VBs decreased their lordotic wedging process during growth, whereas the opposite was observed with the IVDs and SLL, which increased in boys and decreased in girls, becoming independent of sex. The SSS, however, manifested the same process of shape variation, becoming greater in boys than in girls. PMID- 21629162 TI - Comparison of 4 airway devices on cervical spine alignment in a cadaver model with global ligamentous instability at C5-C6. AB - STUDY DESIGN: Human cadaveric study using various intubation devices in a cervical spine instability model. OBJECTIVE: We sought to evaluate various intubation techniques and determine which device results in the least cervical motion in the setting of a global ligamentous instability model. SUMMARY OF BACKGROUND DATA: Many patients presenting with a cervical spine injury have other injuries that may require rapid airway management with endotracheal intubation. Secondary neurologic injuries may occur in these patients because of further displacement at the level of injury, vascular insult, or systemic decrease in oxygen delivery. The most appropriate technique for achieving endotracheal intubation in the patient with a cervical spine injury remains controversial. METHODS: A global ligamentous instability at the C5-C6 vertebral level was created in lightly embalmed cadavers. An electromagnetic motion analysis device (Liberty; Polhemus, Colchester, VT) was used to assess the amount of angular and linear translation in 3 planes during intubation trials with each of 4 devices (Airtraq laryngoscope, lighted stylet, intubating LMA, and Macintosh laryngoscope). The angular motions measured were flexion-extension, axial rotation, and lateral bending. Linear translation was measured in the medial lateral (ML), axial, and anteroposterior planes. Intubation was performed by either an emergency medical technician or by a board-certified attending anesthesiologist. Both time to intubate as well as failure to intubate (after 3 attempts) were recorded. RESULTS: There was no significant difference shown with regards to time to successfully intubate using the various devices. It was shown that the highest failure-to-intubate rate occurred with use of the intubating LMA (ILMA) (23%) versus 0% for the others. In flexion/extension, we were able to demonstrate that the Lightwand (P = 0.005) and Airtraq (P = 0.019) resulted in significantly less angular motion than the Macintosh blade. In anterior/posterior translation, the Lightwand (P = 0.005), Airtraq (P = 0.024), and ILMA (P = 0.021) all caused significantly less linear motion than the Macintosh blade. In axial rotation, the Lightwand (P = 0.017) and Airtraq (P = 0.022) resulted in significantly less angular motion than the Macintosh blade. In axial translation (P = 0.037) and lateral bending (P = 0.003), the Lightwand caused significantly less motion than the Macintosh blade. CONCLUSION: In a cadaver model of C5-C6 instability, the greatest amount of motion was caused by the most commonly used intubation device, the Macintosh blade. Intubation with the Lightwand resulted in significantly less motion in all tested parameters (other than ML translation) as compared with the Macintosh blade. It should also be noted that the Airtraq caused less motion than the Macintoshblade in 3 of the 6 tested planes. There were no significant differences in failure rate or the amount of time it took to successfully intubate in comparing these techniques. We therefore recommend the use of the Lightwand, followed by the Airtraq, in the setting of a presumed unstable cervical spine injury over the Macintosh laryngoscope. PMID- 21629163 TI - Regional variation and spine care: an historical perspective. PMID- 21629164 TI - Clinical outcome of metastatic spinal cord compression treated with surgical excision +/- radiation versus radiation therapy alone: a systematic review of literature. AB - STUDY DESIGN: Systematic literature review from 1970 to 2007. OBJECTIVE: This study reports the results of a systematic review comparing surgical decompression +/- radiation to radiation therapy alone among patients with metastatic spinal cord compression. SUMMARY OF BACKGROUND DATA: Currently, the optimal treatment of metastatic spine lesions is not well defined and is inconsistent. Radiation and surgical excision are both accepted and effective. There appears to be a favorable trend for improved neurological outcome with surgical excision and stabilization as part of the management. METHODS: A review of the English literature from 1970 to 2007 was performed in the Medline database using general MeSH terms. Relevant outcome studies for the treatment of metastatic spinal cord compression were selected through criteria defined a priori. The primary outcome was ambulatory capacity. A random effects model was built to compare results between treatment groups, based on calculated proportions from each study. RESULTS: Of the 1595 articles screened, 33 studies (2495 patients) were selected based on our inclusion and exclusion criteria. Sixty-four percent of the patients who underwent surgical decompression, tumor excision, and stabilization had neurological improvement from nonambulatory to ambulatory status. Twenty-nine percent of the radiation therapy group regained the ability to ambulate after treatment (P < 0.001). Paraplegic patients had a 4-fold greater recovery rate to functional ambulation with surgical intervention than with radiation therapy alone (42% vs. 10%, P < 0.001). Pain relief was noted in 88% of the patients in the surgical studies and in 74% of the patients in studies of radiation therapy (P < 0.001). The overall surgical complication rate was 29%. CONCLUSION: This systematic review suggests that surgical excision of tumor and instrumented stabilization may improve clinical outcomes compared with radiation therapy alone, with regard to neurological function and pain. However, most data in the current literature are from observational studies, where variations in patient population and treatments cannot be controlled. This compromised our ability to compare the results of both treatments directly. PMID- 21629165 TI - The pros and cons of evidence-based medicine. PMID- 21629167 TI - Reliability of the Spinal Deformity Study Group classification of lumbosacral spondylolisthesis. AB - STUDY DESIGN: Reliability study of the computer-assisted SDSG (Spinal Deformity Study Group) classification of lumbosacral spondylolisthesis. OBJECTIVE: To assess the intra- and interobserver reliability of the computer-assisted SDSG classification of lumbosacral spondylolisthesis. SUMMARY OF BACKGROUND DATA: The SDSG has proposed a new classification of lumbosacral spondylolisthesis based on slip grade, pelvic incidence (PI), and sacro-pelvic and spinal balance. Three types of low-grade spondylolisthesis are described: low PI (type 1), normal PI (type 2), and high PI (type 3). High-grade spondylolisthesis are defined as type 4 (balanced sacro-pelvis), type 5 (retroverted sacro-pelvis with balanced spine), and type 6 (retroverted sacro-pelvis with unbalanced spine). METHODS: Full-length standing lateral radiographs of the spine of 40 subjects with lumbosacral spondylolisthesis were reviewed twice by 7 observers. Custom software was used by the observers to identify 7 anatomical landmarks on each radiograph to determine the SDSG type for all subjects. Percentage of agreement and kappa coefficients were used to determine the intra- and interobserver reliability. RESULTS: All 6 types of spondylolisthesis described in the computer-assisted SDSG classification were identified. Overall intra- and interobserver agreements were 80% (kappa: 0.74) and 71% (kappa: 0.65), respectively. The intra- and interobserver agreements associated with computerized determination of slip grade were 92% (kappa: 0.83) and 88% (kappa: 0.78), respectively. As for computerized determination of sacro-pelvic and spinal balance, intra- and interobserver agreements were 86% (kappa: 0.76) and 75% (kappa: 0.63) for low-grade slips, whereas they were 88% (kappa: 0.80) and 83% (kappa: 0.75) for high-grade slips. CONCLUSION: Substantial intra- and interobserver reliability was found for the computer-assisted SDSG classification, and all 6 types of lumbosacral spondylolisthesis were identified. Refinement of the computer-assisted classification technique is, however, needed to further increase the reliability of the SDSG classification and facilitate its clinical use. PMID- 21629168 TI - Assessment of pedicle perforation by the cervical pedicle screw placement using plain radiographs: a comparison with computed tomography. AB - STUDY DESIGN: A retrospective study. OBJECTIVE: To suggest methods for detecting pedicle perforation on the basis of cervical pedicle screw (CPS) position on plain radiographs. SUMMARY OF BACKGROUND DATA: No studies have reported correlations between CPS position and pedicle perforation as observed on plain radiographs. This study was performed under the assumption that the detection of pedicle perforation would help to minimize the risks of neurovascular injury and help to obtain stable fixation. METHODS: A total of 48 subjects (with 205 screws) who had undergone CPS placement from C3 to C7 were enrolled in this study. To evaluate CPS position, the positions of the screw heads (neutral; the lateral margin of lateral mass cross the polyaxial screw head core, medial, or lateral) and tips (medial to uncovertebral joint [UVJ], within UVJ, or lateral to UVJ) on anteroposterior (AP) radiographs were analyzed. On the postoperative computed tomography, we analyzed the grade of pedicle perforation (grade 0: no PF; 1: < 25%; 2: 20%-50%; 3: > 50% of the screw diameter violation). Grades 0 and 1 were considered to be the correct position. RESULTS: Correct positioning was found for 174 screws (84.9%), and incorrect positioning was found for 31 screws (15.1%). The screw head was placed in a neutral position for 182 screws (88.8%), in the lateral position for 15 screws (7.3%), and in the medial position for 8 screws (3.9%). Of the 182 screws whose heads were in neutral position, 151 (83%) screws whose tips were located medial to the UVJ area were correctly positioned (sensitivity 0.89, specificity 1.0). A significant correlation was observed between the position of the screw tip and the grade of pedicle perforation (P = 0.000). CONCLUSION: A screw with a head that is located in a neutral position and a tip that is placed medial to the UVJ area on plain radiographs is considered to be in the safest position. A tip positioned lateral to the UVJ area or a head located out of the neutral position is expected to increase the risk of perforation. The use of intraoperative radiographs during CPS placement will help to identify the screws that are expected to cause pedicle perforation and allow the appropriate corrections to be made. PMID- 21629169 TI - Techniques for operative correction of proximal junctional kyphosis of the upper thoracic spine. AB - STUDY DESIGN: Retrospective study of a consecutive series of patients treated for proximal junctional kyphosis (PJK) of the upper thoracic and cervicothoracic spine. OBJECTIVE: To discuss corrective techniques for the management of symptomatic kyphosis at the junction of fused and mobile segments of the upper thoracic and cervicothoracic spine in patients who complain of pain, neurological deficit, ambulatory difficulty, and/or social isolation. SUMMARY OF BACKGROUND DATA: PJK is an unfortunately common, but important, complication seen in long instrumented fusions to the upper thoracic and cervicothoracic spine. Although often asymptomatic, its incidence and prevalence warrant a discussion on treatment options for symptomatic patients. METHODS: After the institutional review board confirmed approval, we retrospectively analyzed patients who received treatment of PJK from 2003 to 2009. Segmental instrumentation and intraoperative neurophysiological monitoring were used in all patients. Data acquisition was performed by reviewing electronic medical records and radiographs. Inclusion criteria were patients who underwent surgical correction of PJK of the cervicothoracic and upper thoracic spine and had more than 2-year follow-up. Preoperative lumbar lordosis, preoperative thoracic kyphosis, pre- and postoperative sagittal balance, and sagittal proximal junctional Cobb angle were obtained. All corrective procedures were performed in 2 stages, each patient receiving cervical traction between cases. RESULTS: Inclusion criteria were met in 7 patients (5 women and 2 men), with mean age of 55 years (range, 18-80 years). Six patients received multilevel Smith-Petersen osteotomies, with 2 patients receiving rib osteotomies, and 1 patient received a vertebral column resection. The mean preoperative and postoperative proximal junctional Cobb angles were 45 degrees (range, 14 degrees -89.7 degrees ) and 14 degrees (range, 3.0 degrees -38.0 degrees ), respectively. The mean degree of correction was 31 degrees (range, 11 degrees -79.2 degrees ). All patients had maintained or improved sagittal balance. No patient sustained a temporary or permanent neurological deficit after correction related to surgery. All patients had 2-year follow-up, and there were no mortalities. CONCLUSION: For a selected cohort of patients who develop PJK of the upper thoracic and cervicothoracic spine, osteotomies, cervical traction, and intraoperative manual reduction provide a significant improvement of proximal junctional Cobb angles. To our knowledge, this is the first study to address treatment for symptomatic patients with this condition. PMID- 21629170 TI - Kinematics of cervical total disc replacement adjacent to a two-level, straight versus lordotic fusion. AB - STUDY DESIGN: In vitro biomechanical study. OBJECTIVE: To characterize cervical total disc replacement (TDR) kinematics above two-level fusion, and to determine the effect of fusion alignment on TDR response. SUMMARY OF BACKGROUND DATA: Cervical TDR may be a promising alternative for a symptomatic adjacent level after prior multilevel cervical fusion. However, little is known about the TDR kinematics in this setting. METHODS: Eight human cadaveric cervical spines (C2 T1, age: 59 +/- 8.6 years) were tested intact, after simulated two-level fusion (C4-C6) in lordotic alignment and then in straight alignment, and after C3-C4 TDR above the C4-C6 fusion in lordotic and straight alignments. Fusion was simulated using an external fixator apparatus, allowing easy adjustment of C4-C6 fusion alignment, and restoration to intact state upon disassembly. Specimens were tested in flexion-extension using hybrid testing protocols. RESULTS: The external fixator device significantly reduced range of motion (ROM) at C4-C6 to 2.0 +/- 0.6 degrees , a reduction of 89 +/- 3.0% (P < 0.05). Removal of the fusion construct restored the motion response of the spinal segments to their intact state. The C3-C4 TDR resulted in less motion as compared to the intact segment when the disc prosthesis was implanted either as a stand-alone procedure or above a two-level fusion. The decrease in motion of C3-C4 TDR was significant for both lordotic and straight fusions across C4-C6 (P < 0.05). Flexion and extension moments needed to bring the cervical spine to similar C2 motion endpoints significantly increased for the TDR above a two-level fusion compared to TDR alone (P < 0.05). Lordotic fusion required significantly greater flexion moment, whereas straight fusion required significantly greater extension moment (P < 0.05). CONCLUSION: TDR placed adjacent to a two-level fusion is subjected to a more challenging biomechanical environment as compared to a stand-alone TDR. An artificial disc used in such a clinical scenario should be able to accommodate the increased moment loads without causing impingement of its endplates or undue wear during the expected life of the prosthesis. PMID- 21629171 TI - Biomechanical evaluation of an expandable cage in single-segment posterior lumbar interbody fusion. AB - STUDY DESIGN: Controlled laboratory study. OBJECTIVE: To evaluate the biomechanical characteristics of a new expandable interbody cage in single segment posterior lumbar interbody fusion (PLIF) using cadaveric lumbar spines. SUMMARY OF BACKGROUND DATA: One of the popular methods of treating lumbar spine pathologies involves a posterior lumbar interbody fusion using bilateral interbody nonexpandable cages. However, this method can require extensive bony removal and nerve root retraction. Expandable interbody cages may decrease the risk associated with PLIFs. METHODS: Biomechanical testing was performed on 5 fresh frozen L4/L5 mobile functional spinal units using a custom testing system that permits 6 df and a digital video digitizing system. The specimens were tested intact, postdiscectomy, after interbody cage placement, and after cage placement and pedicle screw fixation. Each specimen was tested from 0.5 to 8.0 N.m for extension, flexion, lateral bending, and rotation, and from 5 to 300 N for axial compression. The angular displacement, stiffness, disc height, and sagittal alignment were determined. RESULTS: When the cage was supplemented with pedicle screw fixation, the mean angular displacement for rotation and lateral bending was significantly less than all other conditions (P < 0.05). The percentage range of motion (% ROM) showed a statistically significant decrease in lateral bending (P < 0.05) for cage alone vs. postdiscectomy. For the pedicle screw construct, rotation showed a significantly lower percentage ROM compared with all other constructs (P < 0.05), and lateral bending and extension-flexion showed a significantly lower percentage ROM compared with postdiscectomy (P < 0.05). For all motions, stiffness of the cage and pedicle screw construct was greater than intact, with only rotation showing a statistically significant increase (P < 0.05). Anterior disc height was restored to intact after cage alone (P < 0.05). Sagittal alignment did not show statistically significant differences. CONCLUSION: PLIF using expandable lumbar interbody cage requires pedicle screw fixation. PMID- 21629174 TI - Occipitocervical reconstruction through direct lateral and posterior approach for the treatment of primary osteosarcoma in the atlas: a case report. AB - STUDY DESIGN: Case report. OBJECTIVE: To report a new surgical technique of the primary cervical osteosarcoma in the atlas. SUMMARY OF BACKGROUND DATA: Primary osteosarcoma of the atlas is extremely rare. This is the first report that describes a surgical treatment of the primary osteosarcoma in the atlas. Resection of an osteosarcoma in the atlas is very difficult because of the complex and important anatomic structures that surround it, and secure reconstruction of the atlas is difficult as well. METHODS: A 48-year-old man was referred to our institute with a 10-month history of a palpable painful mass on the right posterior neck. His neck mass was diagnosed as chondroblastic osteosarcoma by open bone biopsy. The plain radiograph of the lateral cervical spine revealed the osteoblastic lesion of the vertebra and an extraosseous mass formation from the C1 to C3 vertebrae. Computed tomography of the cervical spine revealed approximately a 7 * 3 * 7 cm-sized extraosseous calcified mass that originated from the right lateral mass of the atlas. The magnetic resonance imaging of the cervical spine did not show any spinal cord compression. RESULTS: The patient underwent excision of this tumor using the direct lateral approach and reconstruction of the lateral mass of the atlas. On gross examination of the mass, there was a reactive thin membrane ("pseudomembrane") between soft tissue and tumor. At 3 months after surgery, the computed tomographic scan showed the solid fusion state of the occipitocervical joint. He rarely complained of any problems except for mild limitation of neck motion. CONCLUSION: We report a rare case of complete excision of an osteosarcoma of the C1 lateral mass in our patient via a direct lateral and posterior approach to secure additional fixation of occipitocervical joint. We describe our technique for reconstructing the lateral mass of the atlas. This reconstruction technique will also be applicable to other resection surgeries involving the occipitocervical junction. PMID- 21629175 TI - Protection of bile ducts in liver transplantation: looking beyond ischemia. AB - Biliary complications, especially nonanastomotic biliary strictures (NAS), are a major cause of morbidity after orthotopic liver transplantation. Of all donor and recipient characteristics known to increase the risk of developing NAS, the role of prolonged ischemia times is most extensively described in the literature. However, there is increasing evidence that several other, non-ischemia-related factors play a critical role in the pathogenesis of NAS as well. The clinical presentation of NAS may vary considerably among liver transplant recipients, including large variations in time of occurrence, and in location and severity of the strictures. Additional underlying causes such as bile salt toxicity and immune-mediated injury are believed to explain the wide spectrum of biliary strictures after orthotopic liver transplantation. Current and emerging insight in the pathogenesis of NAS and potential targets to reduce biliary injury and preserve bile ducts are discussed in this overview. PMID- 21629176 TI - Adenosine diphosphate receptor P2Y12-mediated migration of host smooth muscle like cells and leukocytes in the development of transplant arteriosclerosis. AB - BACKGROUND: We have recently reported that platelet P2Y12 receptors may play a role in the development of transplant arteriosclerosis (TA). In the present study, we investigated the role of P2Y12 receptors on host-derived smooth muscle like cells (SMLCs, including bone-marrow-derived SMLCs) and CD45+ leukocytes, both of which are believed to be associated with the development of TA, using P2Y12-deficient (KO) mice. METHODS: Orthotopic carotid artery transplantation was performed from C3H/He (H-2k) donors into KO or wild-type (WT) recipient mice (129S:C57BL/6, H-2b). Grafts were harvested at 8 weeks after transplantation for histology. Plasma monocyte chemoattractant protein-1 (MCP-1) levels were analyzed with a kit. Cell migration was examined using a Boyden chamber system. The expression of MCP-1 messenger RNA was assessed by real-time polymerase chain reaction. RESULTS: Eight weeks after allotransplantation, KO recipient mice showed a significant reduction of luminal occlusion, host-derived SMLCs, CD45+ leukocytes, MCP-1+ cells in the grafts, and of plasma MCP-1 levels. In addition, the migration of host-derived SMLCs (including bone-marrow-derived SMLCs) and CD45+ leukocytes stimulated with adenosine diphosphate (ADP) or 2-methylthio-ADP (2MeSADP, a stable ADP analog) was significantly decreased in KO mice. There were no significant changes in MCP-1-induced cell migration between WT and KO mice. The low concentration of 2MeSADP plus MCP-1 significantly increased cell migration in WT but not KO mice. Furthermore, 2MeSADP-induced MCP-1 messenger RNA expression was significantly reduced in the cells of KO mice. CONCLUSIONS: Thus, the P2Y12-mediated migration of host-derived SMLCs and CD45+ leukocytes may play an important role in the development of TA, partly by MCP-1 pathways. PMID- 21629177 TI - Clinical significance of pretransplant chromosomally integrated human herpesvirus 6 in liver transplant recipients. AB - BACKGROUND: The clinical relevance of chromosomally integrated human herpesvirus 6 (CIHHV-6) after transplantation is not known. This study was aimed to determine the potential role of CIHHV-6 on the occurrence of other infections, allograft rejection, and outcomes after liver transplantation. METHODS: A real-time quantitative human herpesvirus-6 (HHV-6) polymerase chain reaction assay was performed on whole blood samples of 548 liver transplant recipients. Clinical characteristics and outcomes of the patients with CIHHV-6 (defined as HHV-6 levels >1 * 10(6) genomes/mL) were compared with those of patients with low-level or no HHV-6 DNAemia. RESULTS: Seven had CIHHV-6, 35 had low-level HHV-6 DNAemia, and 506 had no HHV-6 DNAemia before liver transplantation. Bacterial infection was significantly more common in the CIHHV-6 group compared with the group without HHV-6 (71.4% vs. 31.4%; P = 0.04). A higher rate of allograft rejection was observed in the CIHHV-6 group compared with the group with low-level HHV-6 DNAemia (71.4% vs. 37.1%; P = 0.12) and those without HHV-6 DNAemia (71.4% vs. 42.9%; P = 0.25), although these differences did not reach statistical significance. Other opportunistic infections and outcomes were not significantly different between the CIHHV-6 group and the non-CIHHV-6 groups. CONCLUSION: Patients with CIHHV-6 may be at increased risk of indirect HHV6 effects after transplantation. This clinically relevant observation warrants confirmation using a larger cohort of transplant recipients. PMID- 21629178 TI - Microparticles for diagnosis of graft-versus-host disease after allogeneic stem transplantation. AB - BACKGROUND: The differentiation between acute graft-versus-host disease (aGvHD) and infection is still a clinical challenge in patients after allogeneic hematopoietic stem-cell transplantation (HSCT). Definitive diagnosis is based on histologic findings, but a simple blood test for differentiation is missing. METHODS: In a prospective study, we measured the plasma levels of erythrocyte derived microparticles (EryMP) in 19 recipients during HSCT. Microparticles were isolated by differential centrifugation, double stained for glycophorin A (CD235) and annexin V, and analyzed by flow cytometry. RESULTS: Eight patients developed aGvHD (42%), 15 patients developed infectious complications (79%), and two patients developed microangiopathic hemolytic anemia (11%). The levels of EryMP, as measured before conditioning therapy (535 * 10(6)/L in median), were not affected by total body irradiation, high-dose chemotherapy, or in vivo T-cell depletion. EryMP levels were unaffected in uncomplicated patients during aplasia (522 * 10(6)/L in median; P=0.394) or after engraftment (480 * 10(6)/L in median; P = 0.594) and in patients with infectious complications or sepsis (586 * 10(6)/L in median; P = 0.606). In contrast, in patients who developed aGvHD after HSCT, a 1.7-fold increase in the plasma levels of EryMP was observed (880 *1 0(6)/L in median; P<0.001 compared with the time before therapy and P = 0.015 compared with patients with infections or sepsis). CONCLUSION: Increased plasma levels of EryMP are present in patients who develop aGvHD but not in patients who develop infection or sepsis after HSCT. Therefore, EryMP are a potential, novel, blood marker that may be helpful in the diagnosis of this common complication after HSCT. PMID- 21629180 TI - Locked and unlocked nucleosides in functional nucleic acids. AB - Nucleic acids are able to adopt a plethora of structures, many of which are of interest in therapeutics, bio- or nanotechnology. However, structural and biochemical stability is a major concern which has been addressed by incorporating a range of modifications and nucleoside derivatives. This review summarizes the use of locked nucleic acid (LNA) and un-locked nucleic acid (UNA) monomers in functional nucleic acids such as aptamers, ribozymes, and DNAzymes. PMID- 21629181 TI - Synthesis and antimalarial activity of novel dihydro-artemisinin derivatives. AB - The Plasmodium falciparum cysteine protease falcipain-2, one of the most promising targets for antimalarial drug design, plays a key role in parasite survival as a major peptide hydrolase within the hemoglobin degradation pathway. In this work, a series of novel dihydroartemisinin derivatives based on (thio)semicarbazone scaffold were designed and synthesized as potential falcipain 2 inhibitors. The in vitro biological assay indicated that most of the target compounds showed excellent inhibition activity against P. falciparum falcipain-2, with IC(50) values in the 0.29-10.63 MUM range. Molecular docking studies were performed to investigate the binding affinities and interaction modes for the inhibitors. The preliminary SARs were summarized and could serve as a foundation for further investigation in the development of antimalarial drugs. PMID- 21629182 TI - Phytochemical and cytotoxic investigations of Curcuma mangga rhizomes. AB - Investigations on the cytotoxic effects of the crude methanol and fractionated extracts (hexane, ethyl acetate) C. mangga against six human cancer cell lines, namely the hormone-dependent breast cell line (MCF-7), nasopharyngeal epidermoid cell line (KB), lung cell line (A549), cervical cell line (Ca Ski), colon cell lines (HCT 116 and HT-29), and one non-cancer human fibroblast cell line (MRC-5) were conducted using an in-vitro neutral red cytotoxicity assay. The crude methanol and fractionated extracts (hexane and ethyl acetate) displayed good cytotoxic effects against MCF-7, KB, A549, Ca Ski and HT-29 cell lines, but exerted no damage on the MRC-5 line. Chemical investigation from the hexane and ethyl acetate fractions resulted in the isolation of seven pure compounds, namely (E)-labda-8(17),12-dien-15,16-dial (1), (E)-15,16-bisnor-labda-8(17),11-dien-13 on (2), zerumin A (3), beta-sitosterol, curcumin, demethoxycurcumin and bis demethoxycurcumin. Compounds 1 and 3 exhibited high cytotoxic effects against all six selected cancer cell lines, while compounds 2 showed no anti-proliferative activity on the tested cell lines. Compound 1 also demonstrated strong cytotoxicity against the normal cell line MRC-5. This paper reports for the first time the cytotoxic activities of C. mangga extracts on KB, A549, Ca Ski, HT-29 and MRC-5, and the occurrence of compound 2 and 3 in C. mangga. PMID- 21629183 TI - Microwave assisted synthesis and unusual coupling of some novel pyrido[3,2 f][1,4]thiazepines. AB - 3-Amino-3-thioxopropanamide (1) reacted with ethyl acetoacetate to form 6-hydroxy 4-methyl-2-thioxo-2,3-dihydropyridine-3-carboxamide (2), which reacted with alpha haloketones 3 to produce 2,3-disubstituted-8-hydroxy-6-methyl-2H,5H-pyrido[3,2-f] [1,4]thiazepin-5-ones 4a-c. Benzoylation of 4c led to the formation of the dibenzoate derivative 9. Compounds 4a-c could be prepared stepwise through the formation of S-alkylated derivatives 10a-c. Compounds 2, 4a-c, 9 and 10a-c were prepared using microwave as a source of heat, and gave better yields in shorter times than those achieved by traditional methods. Coupling of 4a-c with arenediazonium chlorides proceeded unusually to give the 6-hydroxy-4-methyl-2 (arylazo)thieno[2,3-b]pyridin-3(2H)-one ring contraction products 14. Structures of the newly synthesized compounds were proven by spectral and chemical methods. PMID- 21629184 TI - High-frequency electrical stimulation in the nucleus accumbens of morphine treated rats suppresses neuronal firing in reward-related brain regions. AB - BACKGROUND: Previous studies have reported that high-frequency stimulation (HFS) in the nucleus accumbens (NAc) is a potential treatment modality for drug craving and relapse. We aimed to explore the electrophysiological changes in reward related brain regions during NAc stimulation and reveal the effects of stimulation frequency and target changes on NAc neuronal activities. MATERIAL/METHODS: Twenty-eight rats were randomized into saline (n=8) and morphine (n=20) groups. The morphine group was further divided into core (n=10, only the core of the NAc was stimulated) and shell (n=10, only the shell of the NAc was stimulated) subgroups. Conditioned place preference (CPP) behavior of the rats was evaluated to confirm morphine preference after morphine injection and CPP training for 10 days. We recorded NAc neuronal responses to NAc core stimulation at different frequencies, as well as changes in VP and VTA neuronal firing during NAc core stimulation, and changes in NAc neuronal firing during NAc shell stimulation. RESULTS: The results indicate that high frequency stimulation was more effective in suppressing NAc neuronal activities than low frequency stimulation and that core stimulation was more effective than shell stimulation. Most VP neurons were inhibited by NAc core stimulation, while VTA neurons were not. CONCLUSIONS: The results suggest that electrical stimulation in the NAc can suppress neuronal firing in reward-related brain regions. The stimulation might be frequency- dependent in suppressing neuronal firing. The core and shell of the NAc play different roles in suppressing NAc neuronal firing as 2 stimulating targets. PMID- 21629185 TI - Differential diagnosis of behavioral variant of fronto-temporal dementia (bvFTD). AB - BACKGROUND: The aim of the paper is the differential diagnosis of various types of Fronto-Temporal Dementia (FTD), with the focus on its behavioural variant (bvFTD). MATERIAL/METHOD: Screening was done in order to assess the depth of dementia with the short version of MMSE, while evaluation of various variants of FTD was performed with the use of such neuropsychological tests as Newcomb and Chicago Fluency Tests, Wechsler Memory Scale-III (WMS-III), Western Aphasia Battery (WAB-R), and the Boston Naming Test (BNT). Behaviour was evaluated with a Polish version of the Frontal Behavioral Inventory (FBInv). The inventory consists of 24 questions which enable an evaluation of social behaviour disorders. The study included 112 patients--68 men and 46 women treated in the Reintegrative-Training Centre of the Foundation for Persons with Brain Dysfunctions in Krakow and in the Clinic for Developmental Psychiatry, Psychotic Disorders and Old Age Psychiatry, of the Medical University at Gdansk, who were suffering from various types of dementia. RESULTS: It was found that FTD patients scored the highest, while the VAD patients scored somewhat lower in the FBInv. At the same time the scores obtained by PPA patients were higher in comparison to the control groups, but not as high as in the case of patients with FTD. In the process of the neurotherapy of FTD patients we found a reduction of the behavioral disturbances, despite the progression of the illness. CONCLUSIONS: The results obtained in the present study confirmed the diagnostic value of FBInv in the differential diagnosis of various types of FTD and in the evaluation of neurotherapy efficacy. PMID- 21629186 TI - The kinetics of mRNA transforming growth factor beta1 expression and its serum concentration in graft-versus-host disease after allogeneic hemopoietic stem cell transplantation for myeloid leukemias. AB - BACKGROUND: Graft-versus-host disease (GVHD) is still a major complication following allogeneic hematopoietic stem cell transplantation (alloHSCT). Recent data indicates that transforming growth factor beta1 (TGF-beta1) may play a role in development of GVH reaction. MATERIAL/METHODS: Forty patients with acute myeloid leukemia (AML) and chronic myeloid leukemia (CML) were included. Quantitative real time polymerase chain reaction (RT-qPCR) was performed to assess the expression of mRNA TGF-beta1. TGF-beta1 serum concentration was assessed using a commercial ELISA. RESULTS: In all patients, a prompt decrease in TGF-beta1 mRNA expression and its serum concentration was demonstrated after conditioning. In patients with acute GVHD, TGF-beta1 mRNA expression and its serum concentration remained low until day +30 after transplant as compared to the day of transplant (p<0.03 and p<0.006, respectively). TGF-beta1 mRNA expression and its serum concentration significantly increased on day +100 in patients who developed chronic GVHD as compared to the day of transplant (p<0.0009 and p<0.02, respectively). CONCLUSIONS: TGF-beta1 seems to be an additional regulator of donor engraftment; its low levels probably being one of the factors contributing to the development of acute GVHD. On the other hand, chronic GVHD symptoms seem to correlate with high TGF-beta1 mRNA expression and its serum concentration in patients who underwent bone marrow transplantation for myeloid leukemias. Nevertheless, further studies with greater numbers of patients are needed to establish the role of TGF-beta1 in graft-versus-host disease pathophysiology. PMID- 21629187 TI - Oxidative damage DNA: 8-oxoGua and 8-oxodG as molecular markers of cancer. AB - BACKGROUND: The broad spectrum of oxidative damage DNA biomarkers: urinary excretion of 8-oxodG (8-oxo-7,8-dihydro-2'-deoxyguanosine), 8-oxoGua (8-oxo-7,8 dihydroguanine) as well as the level of oxidative damage DNA in leukocytes, was analyzed in cancer patients and healthy subjects. MATERIAL/METHODS: 222 cancer patients and 134 healthy volunteers were included in the analysis, using methodologies which involve HPLC (high-performance liquid chromatography) prepurification followed by gas chromatography with isotope dilution mass spectrometry detection and HPLC/EC. RESULTS: For the whole patient population (n=222) the median values of 8-oxoGua and 8-oxodG in urine samples were 12.44 (interquartile range: 8.14-20.33) [nmol/24 hr] and 6.05 (3.12-15.38) [nmol/24 hr], respectively. The median values of 8-oxoGua and 8-oxodG in urine samples of the control group (n=85) were 7.7 (4.65-10.15) [nmol/24 hr] and 2.2 (1.7-2.8) [nmol/24 hr], respectively. The level of 8-oxodG in DNA isolated from leukocytes of the patient population (n=179) and of the control group (n=134) was 4.93 (3.46 9.27) per 10'6 dG and 4.46 (3.82-5.31) per 10'6 dG, respectively. CONCLUSIONS: The results suggest that oxidative stress in cancer patients, demonstrated by augmented amounts of these modifications in urine, could be typical not only for affected tissue but also for other tissues and even the whole organism. An assay that enables the determination of levels of basic markers of oxidative stress might be applied in clinical practice as an additional, helpful marker to diagnose cancer. PMID- 21629188 TI - Low frequency haplotypes of E-selectin polymorphisms G2692A and C1901T give increased protection from coronary artery disease. AB - BACKGROUND: E-selectin polymorphisms are an independent atherosclerosis and coronary artery disease (CAD) risk factor. This study aimed to investigate the link between the C1901T and G2692A E-selectin tagging SNPs and their haplotypes and the extent of coronary artery disease in Polish patients. MATERIAL/METHODS: For this study 321 patients were recruited CAD extent by coronary angiography and E selectin gene variant were investigated using HapMap, PCR/RFLP, multivariate logistic regression and haplotype analysis. RESULTS: Frequency distributions of the C1901T and G2692A polymorphisms were significantly different in CAD patients as compared to control subjects (p=0.037 and p=0.025, respectively). The C1901T polymorphism was found to be an independent genetic predictor of risk of CAD (OR=3.01) in a multivariate model adjusted for classic, environmental risk factors. The A-C and G-T haplotypes showed the strongest significant associations with CAD. The A-C haplotype proved to be significantly more common in controls (haplotype frequency 9.2%) than in CAD (5.7%, p=0.048); the G-T haplotype was not found among control subjects (0.0%) but was found in CAD (1.3%, p=0.0099). CONCLUSIONS: Associations between the C1901T and G2692A E-selectin polymorphisms and CAD in the Polish population were found. Investigated variants correlated with the risk of coronary artery disease development but not with the extent of coronary artery vascular changes. In the haplotype analysis, 2 haplotypes influenced CAD--the A-C haplotype (7%) proved to exert a protective effect against CAD, while the effect of the less frequent G-T haplotype (1%) was associated with significant increase in CAD risk. PMID- 21629189 TI - Pain during external dacryocystorhinostomy with local anesthesia. AB - BACKGROUND: External dacryocystorhinostomy (DCR) is often performed under local anesthesia (LA) without adequate knowledge of the pain experienced by the patient. MATERIAL/METHODS: We subdivided our surgical technique into stages easily understood by the patients (introducing cotton tipped applicators, performing parabulbar injection, creating the incision, bone cracking (opening the ostium), manipulating the nose, intubating, closing the wound, and packing with gauze). A total of 50 patients ranging in age from 31 to 83 years of age (63.64+/-9.64) underwent external DCR. Each patient was asked 30 minutes after surgery to indicate the intensity of pain experienced at each stage of the surgery and during intramuscular (i.m.) injection of an antibiotic using a visual analog scale (VAS). RESULTS: Analysis of the VAS-based pain scores indicated 3 statistically equal occurrences of pain coinciding with the opening of the ostium, and receiving both parabulbar anesthetic and i.m. antibiotic injections. CONCLUSIONS: The level of pain experienced during the most unpleasant stage of external DCR (ostium opening) was similar to the pain experienced from an i.m. injection. Patients can be informed that pain during external DCR with local anesthesia is comparable to receiving an i.m. gluteal injection. PMID- 21629190 TI - A randomized, double-blinded, placebo-controlled pilot study of probiotics in active rheumatoid arthritis. AB - BACKGROUND: To examine the effect of probiotics as adjunctive therapy for the treatment of rheumatoid arthritis (RA). A sample size of 30 subjects was calculated to determine a moderate effect. MATERIAL/METHODS: A three month double blind, placebo-controlled study was performed using probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 capsules administered orally. Inclusion criteria required at least 4 swollen and 4 tender joints and stable medications with no steroids for at least 1 month prior to and during the study. Twenty-nine patients with RA were randomized to treatment. ACR20 responses, serum cytokine levels and safety parameters were assessed. RESULTS: Fifteen patients were randomized to the probiotic group, and 14 to placebo. Three subjects in the probiotic (20%) and one in the placebo group (7%) achieved an ACR20 response (p= 0.33). There was no statistically significant difference between individual components of the ACR20 criteria. Changes in cytokines favored placebo over probiotic. There was a significant improvement in the Health Assessment Questionnaire (HAQ) score in the probiotic group from visit 1 to visit 3 (p=0.02) but no between-group differences. CONCLUSIONS: Due to inclusion criteria, patients selected for the study had stable RA with chronic synovitis, and thus it may have been difficult for an adjunctive therapy to demonstrate improvement within 3 months. Although probiotics did not clinically improve RA as measured by the ACR20, it is interesting that there was functional improvement seen within the probiotic group compared to placebo. PMID- 21629191 TI - A case of meningitis due to Achromobacter xylosoxidans denitrificans 60 years after a cranial trauma. AB - BACKGROUND: Achromobacter xylosoxidans (AX) is a non-fermentative aerobic gram negative bacillus. It is an opportunistic pathogen and the causative agent of various infections. We report an original case of late posttraumatic meningitis due to AX denitrificans. CASE REPORT: An 83-year-old man was hospitalized for acute headache, nausea and vomiting. The emergency brain computer tomography (CT) scan did not reveal any anomaly. In his medical history, there was an auditory injury due to a cranial trauma incurred in a skiing accident 60 years earlier. Cytobiochemical analysis of the cerebrospinal fluid (CSF) revealed increased levels of neutrophils and proteins. The CSF bacterial culture was positive: the Gram stain showed a gram-negative bacillus, oxidase + and catalase +, and the biochemical pattern using the API 20 NE strip revealed AX dentrificans. Late posttraumatic meningitis on a possible osteomeningeal breach was diagnosed even though the breach was not confirmed because the patient declined a second brain CT scan. The patient was successfully treated with meropenem. CONCLUSIONS: This report demonstrates the importance of searching for unusual or atypical organisms when the clinician encounters meningitis in a particular context, as well as the importance of adequate follow-up of craniofacial traumas. PMID- 21629192 TI - T cell lymphoma presenting as esophageal obstruction and bronchoesophageal fistula. AB - BACKGROUND: The mediastinum is an uncommon location for presentation of peripheral T cell lymphoma. Esophageal involvement by non-Hodgkin's lymphoma is extremely unusual. Although staging can be performed with routine imaging studies, surgical intervention is often required to ensure accurate histologic diagnosis of these lymphomas. Peripheral T cell lymphomas not otherwise specified are among the most aggressive non-Hodgkin lymphomas with often a poor response to conventional chemotherapy. CASE REPORT: We report a case of a 63 year-old-man with an aggressive mediastinal T cell lymphoma presenting as esophageal obstruction and bronchoesophageal fistula. The patient was treated with a cyclophosphamide, vincristine, and prednisone (COP) regimen. Repeat computer tomography scan of the chest after chemotherapy noted a significant decrease in the cavitary lesion in the right paraesophageal region and right mediastinum. Bronchoscopy revealed a large opening in the posterior wall of the bronchus intermedius leading into the esophagus. A fistulogram was done which clearly demonstrated a fistulous tract between the lower esophagus and the right intermediate bronchus secondary to perforation from the lymphoma. The patient eventually underwent cervical esophagostomy and jejunostomy tube placement to correct the brochoesophageal fistula. CONCLUSIONS: The mediastinum is an uncommon location for presentation of peripheral T cell lymphomas, and surgical intervention is often required to ensure accurate histological diagnosis of these lymphomas. In our patient, aggressive mediastinal T cell lymphoma presented as esophageal obstruction and bronchoesophageal fistula. PMID- 21629193 TI - Anterior ischemic optic neuropathy after conventional coronary artery bypass graft surgery. AB - BACKGROUND: Perioperative optic neuropathy is a disease which can lead to serious, irreversible damage of vision. This complication could be the result of non-ocular surgery, for example, cardiac or spinal procedures. We present a case of anterior ischemic neuropathy (AION) which occurred following a conventional coronary artery bypass graft procedure. CASE REPORT: A 57-year-old man, 4 days after Conventional Coronary Artery Bypass Graft surgery as result of multi-vessel stabile coronary artery disease and history of anterolateral wall myocardial infarction, was admitted to the Eye Clinic due to significant loss of vision in his right eye. The patient had hypertension and was a heavy smoker. On admission, the slit lamp examination revealed a relative afferent pupillary defect in the right eye. The fundus examination showed optic disc edema with the presence of flame hemorrhages. Best corrected visual acuity (BCVA) was 0.02. The results of eye examination and fluorescein angiography confirmed the diagnosis of AION. Anti aggregation and antithrombotic treatment was continued with steroids and vasodilators. After 7 days of this treatment we noticed the improvement of BCVA to 0.2. At 6-month follow-up, the vision was stable, and fundus examination revealed optic disc atrophy. CONCLUSIONS: After cardiac surgical operations, such as coronary artery bypass graft procedures, anterior ischemic optic neuropathy may occur. In those cases, close cooperation between the various specialists is necessary. PMID- 21629194 TI - Severe decrease in visual acuity with choroidal hypoperfusion after photodynamic therapy. AB - BACKGROUND: Photodynamic therapy (PDT) is considered a selective method of treatment which works in areas of choroidal neovascularization (CNV); however, there are reports of choroidal hypoperfusion after PDT. This paper presents a clinical case of choroidal circulation disturbances caused by PDT, accompanied by CNV progression. CASE REPORT: The patient, a 75-year-old woman, was qualified for PDT in the right eye--first treatment due to progression of occult CNV. Best corrected visual acuity (BCVA) in the right eye at baseline was +0.3 logMAR. After PDT, a rapid decrease in visual acuity to +0.7 logMAR in the right eye was observed, central choroidal hypoperfusion in fluorescein angiography (FA) with subretinal fluid appeared and, as a consequence, progression of neovascular age related macular degeneration (AMD). After stabilizing the local state through conservative therapy, a decision was made to treat the right eye with intravitreal injections of vascular endothelial growth factor (VEGF) inhibitor. During a 12-month period of observation, 7 doses of ranibizumab were administered. A regression in activity of wet AMD was observed, with visual acuity of +0.6 logMAR. CONCLUSIONS: Choroidal circulation disturbance after PDT is possible and has to be taken into account. Sporadically, it can lead to an acute decrease in visual acuity and local state. After stabilization of AF and optical coherence tomography imaging, further treatment of neovascular AMD with intravitreal injections of anti-VEGF agents should be considered. PMID- 21629195 TI - Activated protein C inhalation: a novel therapeutic strategy for acute lung injury. AB - Acute lung injury (ALI) is a critical illness syndrome with a mortality rate of 25-40%. Despite recent advances of our understanding of the pathophysiology of ALI, no pharmacologic therapies have been proven effective. The key pathogenesis of ALI is the activation of the coagulation cascade and impaired fibrinolysis, resulting in extensive fibrin and hyaline membrane deposition. Activated protein C (APC), an endogenous protein that promotes fibrinolysis and inhibits thrombosis, can modulate the coagulation and inflammation associated with ALI. It is therefore reasonable to suggest that preventing the progression of pulmonary coagulopathy, by restoring normal intraalveolar levels of protein C, will be of therapeutic benefit to patients with ALI. However, a recent clinical trial demonstrated that APC did not improve outcomes from ALI, raising the possibility that the method of APC administration, intravenous infusion or inhalation, may influence the outcomes. In this article we propose the hypothesis that APC inhalation might be a promising and novel choice in the treatment of ALI. PMID- 21629197 TI - Post-processing analysis of transient-evoked otoacoustic emissions to detect 4 kHz-notch hearing impairment--a pilot study. AB - BACKGROUND: To identify a parameter to distinguish normal hearing from hearing impairment in the early stages. The parameter was obtained from transient-evoked otoacoustic emissions (TEOAEs), overcoming the limitations of the usually adopted waveform descriptive parameters which may fail in standard clinical screenings. MATERIAL/METHODS: Audiometric examinations and TEOAE analysis were conducted on 15 normal ears and on 14 hearing-impaired ears that exhibited an audiometric notch around 4 kHz. TEOAE signals were analyzed through a multivariate technique to filter out the individual variability and to highlight the dynamic structure of the signals. The new parameter (named radius 2-dimension--RAD2D) was defined and evaluated for simulated TEOAE signals modeling a different amount of hearing impairment. RESULTS: Audiometric examinations indicated 14 ears as impaired hearing (IH), while the TEOAE ILO92 whole reproducibility parameter (WWR) indicated as IH 7 signals out of 14 (50%). The proposed new parameter indicated as IH 9 signals out of 14 (64%), reducing the number of false negative cases of WWR. CONCLUSIONS: In this preliminary study there is evidence that the new parameter RAD2D defines the topology and the quantification of the damage in the inner ear. The proposed protocol can be useful in hearing screenings to identify hearing impairments much earlier than conventional pure tone audiometry and TEOAE pass/fail test. PMID- 21629196 TI - Modulation of vitamin D signaling is a potential therapeutic target to lower cardiovascular risk in chronic kidney disease. AB - While it is true that many traditional cardiovascular risk factors are amenable to intervention in chronic kidney disease (CKD), the results of intervention may not be as efficacious as those obtained in the general population. Thus, there may also be a unique milieu established in CKD, which causes excess cardiovascular disease (CVD) burden by mechanisms that are as yet not fully recognized. Recently, vitamin D has sparked widespread interest because of its potential favorable benefits on CVD. However, the mechanisms for how vitamin D may improve CVD risk markers and outcomes have not been fully elucidated. Furthermore, hypovitaminosis D is highly prevalent in the CKD cohort. Given this background, we hypothesize that low vitamin D status may act as a new CVD risk marker, and modulation of vitamin D signaling may be a potential therapeutic target to lower cardiovascular risk in CKD. The data presented in this review support that the low vitamin D status may be linked with the high cardiovascular risk in CKD, based on both the biological effects of vitamin D itself on the cardiovascular system, and the cross-actions between vitamin D signaling and the multiple metabolic pathways. Considering the high prevalence of hypovitaminosis D, limited natural vitamin D food sources, and reduced sun exposure in CKD patients, recommendations for treatment of hypovitaminosis D mainly focus on exogenous supplementation with vitamin D and its analogues. Although promising, when to start therapy, the route of administration, the dose, and the duration remain need to be discussed. PMID- 21629198 TI - Predicting the adult height of girls with central precocious puberty. AB - BACKGROUND: There are no absolute criteria for identifying those girls with idiopathic central precocious puberty (CPP) who will benefit from gonadotropin releasing hormone analog (GnRHa) treatment. Our objective was to predict at initial evaluation the differences between adult height (AH) and target height (TH) and (for untreated girls) the time between puberty onset and first menstruation. MATERIAL/METHODS: The 122 girls with CPP who reached their AH included 70 who were given GnRHa because their predicted AH was <155 cm (n=24), their luteinising hormone (LH)/follicle-stimulating hormone peaks (FSH) ratio was >0.66 (n=41) and/or their estradiol was >15 pg/ml (n=40). The other 52 were untreated because their predicted AH was >155 cm. Multiple linear regressions were performed on several subsets of variables. RESULTS: Treated: the difference between AH and TH (-0.6+/-5.4 cm) was predicted by (using SDS) =3.68 (height at initial evaluation - TH) - 1.94 (height at initial evaluation-predicted AH) - 4.23; R2=0.73. Untreated: the difference between AH and TH (1.7+/-4.3 cm) was predicted by =2.76 (height at initial evaluation - TH) - 3.68 LH/FSH peaks ratio 3.49; R2=0.77. Time between puberty onset and first menstruation (years) was predicted by =12.2 - 1.06 age CPP - 0.4 (height at initial evaluation - TH); R2=0.75. CONCLUSIONS: A greater difference between height at initial evaluation and TH (SDS) is associated with a greater AH in treated and untreated girls, as are smaller differences between height at initial evaluation and predicted AH in treated and lower LH/FSH peaks ratios in untreated girls. PMID- 21629199 TI - Underlying diseases in syncope of children in China. AB - BACKGROUND: Syncope accounts for about 1-2% of emergency department visits, but the etiology in many patients with syncope is unclear. Recently, with the use of the head-up tilt test (HUT), the number of patients with unexplained syncope (UPS) has been decreasing; however, the spectrum of underlying diseases of syncope in children is unclear. This retrospective study aimed to analyze the spectrum of underlying diseases in children with syncope. MATERIAL/METHODS: This multi-center clinical study consisted of 888 children (417 males, 471 females, aged 5-18 yrs, median age 12.0+/-3.0 yrs) with syncope who came from Beijing city, Hunan province, Hubei province and Shanghai from August 1999 to March 2009. The clinical and laboratory data of children were studied and the spectrum of underlying diseases in children with syncope was analyzed. RESULTS: In 888 children with syncope, 175 (19.7%) had vasovagal syncope (VVS) with vasoinhibitory response, 35 (3.9%) had VVS with cardioinhibitory response, 73 (8.2%) had VVS with mixed response, 286 (32.2%) had postural orthostatic tachycardia syndrome (POTS), 19 (2.1%) had orthostatic hypotension, 7 (0.9%) had situational syncope, 13 (1.5%) had cardiogenic syncope, and 280 (31.5%) had unexplained syncope. CONCLUSIONS: The data suggest that neurally-mediated syncope was the most common cause in children with syncope. POTS and VVS were the most common hemodynamic patterns of neurally-mediated syncope. PMID- 21629200 TI - A prospective double-blind, randomized clinical trial of levocarnitine to treat autism spectrum disorders. AB - BACKGROUND: L-carnitine was proposed as a potential treatment for patients diagnosed with an autism spectrum disorder to improve mitochondrial dysfunction, but no prior randomized controlled trials have been conducted. MATERIAL/METHODS: Thirty subjects diagnosed with an ASD were randomly assigned to receive a standardized regimen (50 mg L-carnitine/kg bodyweight/day) of liquid L-carnitine (n=19) or placebo (n=11) for 3-months. Measures included changes in professionally completed Childhood Autism Rating Scale (CARS), hand muscle testing, and modified clinical global impression (CGI) forms; parent completed Autism Treatment Evaluation Checklist (ATEC), treatment adherence measurement (TAM), frequency and intensity of side effect rating (FISER)/global rating of side effect burden (GRSEB)/patient report of incidence of side effects (PRISE) forms; and lab testing. RESULTS: Significant improvements were observed in CARS ( 2.03, 95% CI=-3.7 to -0.31), CGI (-0.69, 95% CI=-1.1 to -0.06), and ATEC scores. Significant correlations between changes in serum free-carnitine levels and positive clinical changes were observed for hand muscle strength (R2=0.23, P=0.046), cognitive scores (R2=0.27, P=0.019), and CARS scores (R2=0.20, P=0.047). Study subjects were protocol-compliant (average adherence was >85%) and generally well-tolerated the L-carnitine therapy given. CONCLUSIONS: L-carnitine therapy (50 mg/kilogram-bodyweight/day) administered for 3-months significantly improved several clinical measurements of ASD severity, but subsequent studies are recommended. PMID- 21629201 TI - The relationship between levels of von Willebrand factor and mean platelet volume in subjects with isolated impaired fasting glucose. AB - BACKGROUND: To understand the heterogeneity of platelets, we investigated the correlation between von Willebrand factor (vWf) and mean platelet volume (MPV) in subjects with isolated impaired fasting glucose (IFG). MATERIAL/METHODS: We selected 48 subjects with isolated IFG and 48 normoglycemic control subjects matched for age, sex, and body mass index. We measured levels of vWf and MPV in all subjects. RESULTS: The levels of vWf and MPV were significantly higher in the isolated IFG group than the control group (p<0.05) Also, vWf level was positively correlated with MPV level in subjects in the isolated IFG group (r=0.452, p=0.001). CONCLUSIONS: Our results suggest that vWf seems to be profoundly related to platelet volume in subjects with isolated IFG. PMID- 21629202 TI - Heart rhythm turbulence and NT-proBNP in decompensated liver cirrhosis--a pilot study. AB - BACKGROUND: Heart rhythm turbulence (HRT) is a novel tool for evaluation of cardiovascular mortality. Liver cirrhosis is associated with hemodynamic and myocardial disturbances termed cirrhotic cardiomyopathy. In the stable stage of liver cirrhosis, systolic and myocardial dysfunction is correlated with brain natriuretic peptide (BNP). The aim was to evaluate HRT and its correlation with NT-proBNP, echocardiographic and biochemical parameters in patients with decompensation of liver cirrhosis. MATERIAL/METHODS: The study included 18 patients with decompensated liver cirrhosis and 18 healthy volunteers. Participants underwent echocardiography and 24-hour ECG monitoring. Serum NT proBNP and other biochemical parameters were measured. Turbulence onset (TO) and turbulence slope (TS) were used to indicate HRT. RESULTS: Mean HR (87/min vs. 75/min), TO (-0.385% vs. -0.92%), NT-proBNP (304.85 pg/ml vs. 83.2 pg/ml), LAd (42.5 mm vs. 34.5 mm), RVdd (29.5 mm vs. 25 mm), SPAP (36.5 mmHg vs. 22.5 mmHg) were significantly (p<0.05) higher in patients with liver cirrhosis. Patients with normal TO and TS had better stage in Child-Pugh classification (P=0.04) than patients with abnormal values. Significant negative correlation was found between creatinine and TO, and between mean HR and TS, and significant positive correlation was found between LAd and TS. LV diastolic dysfunction was noted in a majority of cirrhotic patients (n=16). CONCLUSIONS: Patients with decompensated cirrhosis had elevated levels of NT-proBNP and LV diastolic dysfunction. TO values in cirrhotic patients differed significantly from the control group. These findings can indicate risk of symptomatic heart failure development and may be a marker of cirrhotic cardiomyopathy. HRT parameters seem not to be appropriate death predicators. PMID- 21629203 TI - Takotsubo cardiomyopathy--a clinical review. AB - Stress cardiomyopathy is characterised by reversible left ventricular dysfunction. It simulates an acute coronary syndrome (ACS), presenting with precordial pain or dyspnoea, changes of the ST segment, T wave, or QTc interval on electrocardiogram, and raised cardiac enzymes. Typical findings are disturbances of segmental contractility (apical hypokinesia or akinesia), with normal epicardial coronary arteries. The true prevalence is unknown, as the syndrome may be under-diagnosed; it is more common in postmenopausal women. There is usually a trigger in the form of physical or psychological stress. The electrocardiographic, echocardiographic, and ventriculographic changes resolve spontaneously over a variable period of time (from days to months). There are a number of pathophysiological theories, none of which has been shown to be definitive, suggesting that all of them may be involved to some extent. The prognosis is generally favourable, and recurrence is very rare. PMID- 21629204 TI - Influenza virus and cell signaling pathways. AB - Influenza viruses comprise a major class of human respiratory pathogens, responsible for causing morbidity and mortality worldwide. Influenza A virus, due to its segmented RNA genome, is highly subject to mutation, resulting in rapid formation of variants. During influenza infection, viral proteins interact with host proteins and exploit a variety of cellular pathways for their own benefit. Influenza virus inhibits the synthesis of these cellular proteins and facilitates expression of its own proteins for viral transcription and replication. Infected cell pathways are hijacked by an array of intracellular signaling cascades such as NF-kappaB signaling, PI3K/Akt pathway, MAPK pathway, PKC/PKR signaling and TLR/RIG-I signaling cascades. This review presents a research update on the subject and discusses the impact of influenza viral infection on these cell signaling pathways. PMID- 21629205 TI - The Short Stay Unit as a new option for hospitals: a review of the scientific literature. AB - BACKGROUND: The short stay unit (SSU) is a ward providing targeted care for patients requiring brief hospitalization and dischargeable as soon as clinical conditions are resolved. Therefore, SSU is an alternative to the ordinary ward (OW) for the treatment of selected patients. The SSU model has been tested in only a few hospitals, and the literature lacks systematic evaluation of the impact of SSU use. The aim of our study was to evaluate the use of SSUs in terms of length of hospital stay, mortality and readmission rate. MATERIAL/METHODS: A random effect meta-analysis was carried out by consulting electronic databases. Studies were selected that focused on comparison between use of SSUs and OWs. Mean difference of length of stay was calculated within 95% confidence intervals. RESULTS: Six articles were selected, for a total of 21 264 patients. The estimated mean difference was -3.06 days (95% CI -4.71, -1.40) in favor of the SSU. The selected articles did not show any differences in terms of mortality and readmission rate. CONCLUSIONS: Use of SSUs could reduce patient length of stay in hospital, representing an alternative to the ordinary ward for selected patients. A shorter period of hospitalization could reduce the risk of hospital-acquired infections, increase patient satisfaction and yield more efficient use of hospital beds. Findings of this study are useful for institutional, managerial and clinical decision-makers regarding the implementation of the SSU in a hospital setting, and for better management of continuity of care. PMID- 21629206 TI - Intranasal application of the melanocortin 4 receptor agonist MSH/ACTH(4-10) in humans causes lipolysis in white adipose tissue. AB - OBJECTIVE: The melanocortin system has a highly significant role in the hypothalamic regulation of body weight and energy expenditure. In animals, intracerebroventricular infusion of melanocortin receptor 4 (MCR-4) agonists increases basal metabolic rate through activation of the sympathetic nervous system and subsequently reduces food intake. In humans, direct access of MCR-4 agonists to the central nervous system can be achieved by a transnasal route, which leads to weight loss with chronic administration. In the present study, we aimed at investigating the effects of intranasally administered MC4-R agonist MSH/ACTH(4-10) on lipolysis and sympathetic nervous system activity in healthy humans. DESIGN: Healthy normal weight, male volunteers (n=10) received either 10 mg MSH/ACTH(4-10) or placebo intranasally in a double-blinded randomized crossover design. Interstitial glycerol release was assessed by microdialysis in abdominal white adipose tissue (WAT) and in skeletal muscle (SM) of the forearm. Local blood flow, systemic blood pressure, heart rate and muscle sympathetic nerve activity (MSNA) within the superficial peroneal nerve were recorded at rest and after nitroprusside infusion. RESULTS: At 45 min after MSH/ACTH(4-10) administration WAT glycerol concentrations increased by 53.4+/-19.3% compared with baseline conditions (P<0.05) and remained significantly higher throughout the experiment when compared with placebo (P<0.05) while local glycerol release in SM was not significantly affected. Resting MSNA was not altered by MSH/ACTH(4 10) administration; however, sympathoexcitation by intravenous nitroprusside was markedly elevated (MSH/ACTH(4-10) 569+/-69% increase to baseline; placebo: 315+/ 64%; P<0.01). CONCLUSION: Intranasally administered MCR-4 agonist MSH/ACTH 4-10 increases both subcutaneous WAT lipolysis and MSNA, which suggests a direct central nervous peptide effect in humans on key factors of human energy metabolism. PMID- 21629207 TI - Correlates of sedentary behavior in 7 to 9-year-old French children are dependent on maternal weight status. AB - OBJECTIVE: Sedentary behavior accounts for overweight and obesity, independently of physical activity. Correlates of sedentary behavior have not been extensively reported in the literature. Our objective was to determine factors associated with such behavior in 7 to 9-year-old French children in 2007, and to assess interactions between the identified correlates. DESIGN: A nationally representative sample of 2525 children participated in the study. Television viewing, video/computer duration and characteristics of the children and their parents were assessed using a questionnaire completed by the parents. Correlates of television viewing (<2 h per day versus >= 2 h per day) as a proxy for sedentary behavior were estimated using multivariate logistic regression. RESULTS: On an average, children spent more than 2 h per day in front of a screen (television: 1 h 32 min (s.e.m.: +/- 0 h 02 min); video/computer: 0 h 40 min (+/- 0 h 02 min)). Television viewing duration was associated with sociodemographic (child's age, weight status, socio-economic characteristics of the family) and behavioral factors (physical and lifestyle activities). In children of non overweight mothers, risk of spending >= 2 h per day in front of a television was significantly higher in those over 9 (versus 7 years: odds ratio (OR): 2.07; 95% confidence intervals (CI): 1.04-4.11), living in an educational priority zone (OR: 1.62; 95% CI: 1.08-2.44), who were not members of a sports team (OR: 2.24; 95% CI: 1.47-3.41), nor declared active by parents (OR: 1.92; 95% CI: 1.13-3.25), and whose parents' education level was lower than high school (OR: 1.84; 95% CI: 1.24-2.72). In contrast, in children of overweight mothers, only the criteria of >= 4 children in the family (versus 2-3 children: OR: 1.87; 95% CI: 1.05-3.35) and no reported parental occupation (versus manager or white collar: OR: 0.29; 95% CI: 0.11-0.76) were associated with watching television >= 2 h per day. CONCLUSIONS: Correlates of sedentary behavior in 7 to 9-year-old children vary according to maternal overweight. Maternal body mass index must therefore be taken into account when developing strategies to prevent a sedentary lifestyle in children. PMID- 21629208 TI - Atrial fibrillation: RF needle improves septal puncture. PMID- 21629209 TI - Heart failure: rolofylline fails to improve renal function in patients with acute HF. PMID- 21629210 TI - Anemia and iron deficiency in heart failure: mechanisms and therapeutic approaches. AB - Anemia and iron deficiency are common in patients with heart failure (HF), and are associated with worse symptoms and adverse outcomes in this population. Although the two can occur together, anemia in HF is often not caused by iron deficiency, and iron deficiency can be present without causing anemia. Erythropoiesis-stimulating agents have been investigated extensively in the past few years and might be of benefit in patients with HF and anemia. However, concerns have arisen regarding the safety of erythropoiesis-stimulating agents in patients with chronic kidney disease and so the results of a large mortality trial are eagerly awaited to provide information on safety in patients with HF. Iron supplementation or replacement is a much older treatment option for patients with HF and anemia, but questions about the safety of intravenous iron, and absorption problems with oral formulations have prevented its widespread use to date. In the past few years, however, new data on the importance of iron deficiency in HF have become available, and a number of studies with intravenous iron have shown promising results. Therefore, this treatment approach is likely to become an attractive option for patients with HF and iron deficiency, both with and without anemia. PMID- 21629212 TI - Heart failure: furosemide--to DOSE or not to DOSE, that is the question. PMID- 21629211 TI - Peripheral artery disease. Part 1: clinical evaluation and noninvasive diagnosis. AB - Peripheral artery disease (PAD) is a marker of systemic atherosclerosis. Most patients with PAD also have concomitant coronary artery disease (CAD), and a large burden of morbidity and mortality in patients with PAD is related to myocardial infarction, ischemic stroke, and cardiovascular death. PAD patients without clinical evidence of CAD have the same relative risk of death from cardiac or cerebrovascular causes as those diagnosed with prior CAD, consistent with the systemic nature of the disease. The same risk factors that contribute to CAD and cerebrovascular disease also lead to the development of PAD. Because of the high prevalence of asymptomatic disease and because only a small percentage of PAD patients present with classic claudication, PAD is frequently underdiagnosed and thus undertreated. Health care providers may have difficulty differentiating PAD from other diseases affecting the limb, such as arthritis, spinal stenosis or venous disease. In Part 1 of this Review, we explain the epidemiology of and risk factors for PAD, and discuss the clinical presentation and diagnostic evaluation of patients with this condition. PMID- 21629213 TI - Optimal therapy for acute lymphoblastic leukemia in adolescents and young adults. AB - Although the survival rate for adolescents and young adults (AYA) with acute lymphoblastic leukemia (ALL) has steadily improved over the past several decades, it still lags behind that of younger children. This Review explores the reasons for this discrepancy and potential solutions, focusing on patients aged 15-22 years. Recent studies that compared the outcome of AYA patients with ALL treated on pediatric or adult clinical trials have shown substantially better outcomes for this patient population obtained with the pediatric trials. Excellent early results have been obtained for patients with ALL aged up to 40-60 years who were treated in adult study groups with pediatric-based regimens. Targeting biological and socio-political features unique to AYA ALL has reduced the 'AYA gap' and has provided the foundation for basic science and translational and clinical AYA initiatives that are charged with the task of discovering further methods to improve the outcome of AYA with ALL. PMID- 21629214 TI - P5 medicine: a plus for a personalized approach to oncology. PMID- 21629215 TI - Combination therapy: New treatment paradigm for locally advanced cervical cancer? PMID- 21629217 TI - Genetics: Melanoma-GRIN and TRAPP it. PMID- 21629216 TI - Antiangiogenic therapy, hypoxia, and metastasis: risky liaisons, or not? AB - All human cells, including cancer cells, need oxygen and nutrients to survive. A widely used strategy to combat cancer is therefore the starvation of tumor cells by cutting off the blood supply of tumors. Clinical experience indeed shows that tumor progression can be delayed by anti-angiogenic agents. However, emerging evidence indicates that in certain experimental conditions, hypoxia as a result of pruning of the tumor microvasculature can promote tumor invasion and metastasis, although these findings are contextual and debated. Genetic studies in mice unveiled that vascular-targeting strategies that avoid aggravation of tumor hypoxia or even promote tumor oxygenation might prevent such an invasive metastatic switch. In this article, we will discuss the emerging link between hypoxia signaling and the various steps of metastasis. PMID- 21629219 TI - Prostate cancer: searching for bone metastases--how, when and why? PMID- 21629218 TI - Targeting 5alpha-reductase for prostate cancer prevention and treatment. AB - Testosterone is the most abundant circulating androgen, and can be converted to dihydrotestosterone (DHT), a more potent androgen, by the 5alpha-reductase enzymes in target tissues. Current treatments for prostate cancer consist of reducing androgen levels by chemical or surgical castration or pure antiandrogen therapy that directly targets the androgen receptor (AR). Although these therapies reduce tumor burden and AR activity, the cancer inevitably recurs within 18-30 months. An approach targeting the androgen-AR axis at different levels could, therefore, improve the efficacy of prostate cancer therapy. Inhibition of 5alpha-reductase is one such approach; however, the two largest trials to investigate the use of the 5alpha-reductase inhibitors (5ARIs) finasteride and dutasteride in patients with prostate cancer have shown that, although the incidence of cancer was reduced by 5ARI treatment, those cancers that were detected were more aggressive than in patients treated with placebo. Thus, the best practice for using these drugs to prevent and treat prostate cancer remains unclear. PMID- 21629220 TI - Differing levels of testosterone and the prostate: a physiological interplay. AB - The controversies surrounding testosterone replacement therapy (TRT) have been addressed in the past few years. Although the androgenic effects of TRT on normal and malignant prostate cells have been studied for over 70 years, little clinical prospective research exists on the physiological responses of prostate tissues to a wide range of serum testosterone levels. The prostate is both an androgen dependent and an androgen-sensitive organ; active processes are triggered at a 'threshold' or 'saturation' level of testosterone. Despite decades of research, no compelling evidence exists that increasing testosterone beyond this threshold level has a causative role in prostate cancer, or indeed changes the biology of the prostate. Testosterone deficiency has marked physiological and clinical effects on men in middle age and beyond. With subnormal testosterone levels, the potential positive benefits of TRT on factors such as muscle mass, libido or erectile function are likely a dose-response phenomenon, and should be considered differently than the threshold influence on the prostate. This Review will re examine classic androgen research and reflect on whether testosterone actually stimulates prostatic cellular growth and progression in a 'threshold' or a 'dose response' (or both) manner, as well as discuss the influence of testosterone on prostate cells in the hypogonadal and eugonadal states. PMID- 21629222 TI - Mesoporous silica nanoparticles as drug delivery systems for targeted inhibition of Notch signaling in cancer. AB - Notch signaling, a key regulator of stem cells, is frequently overactivated in cancer. It is often linked to aggressive forms of cancer, evading standard treatment highlighting Notch as an exciting therapeutic target. Notch is in principle "druggable" by gamma-secretase inhibitors (GSIs), inhibitory peptides and antibodies, but clinical use of Notch inhibitors is restricted by severe side effects and there is a demand for alternative cancer-targeted therapy. Here, we present a novel approach, using imagable mesoporous silica nanoparticles (MSNPs) as vehicles for targeted delivery of GSIs to block Notch signaling. Drug-loaded particles conjugated to targeting ligands induced cell-specific inhibition of Notch activity in vitro and exhibited enhanced tumor retainment with significantly improved Notch inhibition and therapeutic outcome in vivo. Oral administration of GSI-MSNPs controlled Notch activity in intestinal stem cells further supporting the in vivo applicability of MSNPs for GSI delivery. MSNPs showed tumor accumulation and targeting after systemic administration. MSNPs were biocompatible, and particles not retained within the tumors, were degraded and eliminated mainly by renal excretion. The data highlights MSNPs as an attractive platform for targeted drug delivery of anticancer drugs with otherwise restricted clinical application, and as interesting constituents in the quest for more refined Notch therapies. PMID- 21629221 TI - Engineering biomaterial systems to enhance viral vector gene delivery. AB - Integrating viral gene delivery with engineered biomaterials is a promising strategy to overcome a number of challenges associated with virus-mediated gene delivery, including inefficient delivery to specific cell types, limited tropism, spread of vectors to distant sites, and immune responses. Viral vectors can be combined with biomaterials either through encapsulation within the material or immobilization onto a material surface. Subsequent biomaterial-based delivery can increase the vector's residence time within the target site, thereby potentially providing localized delivery, enhancing transduction, and extending the duration of gene expression. Alternatively, physical or chemical modification of viral vectors with biomaterials can be employed to modulate the tropism of viruses or reduce inflammatory and immune responses, both of which may benefit transduction. This review describes strategies to promote viral gene delivery technologies using biomaterials, potentially providing opportunities for numerous applications of gene therapy to inherited or acquired disorders, infectious disease, and regenerative medicine. PMID- 21629223 TI - Anti-CD22 antibody targeting of pH-responsive micelles enhances small interfering RNA delivery and gene silencing in lymphoma cells. AB - The application of small interfering RNA (siRNA) for cancer treatment is a promising strategy currently being explored in early phase clinical trials. However, efficient systemic delivery limits clinical implementation. We developed and tested a novel delivery system comprised of (i) an internalizing streptavidin conjugated monoclonal antibody (mAb-SA) directed against CD22 and (ii) a biotinylated diblock copolymer containing both a positively charged siRNA condensing block and a pH-responsive block to facilitate endosome release. The modular design of the carrier facilitates the exchange of different targeting moieties and siRNAs to permit its usage in a variety of tumor types. The polymer was synthesized using the reversible addition fragmentation chain transfer (RAFT) technique and formed micelles capable of binding siRNA and mAb-SA. A hemolysis assay confirmed the predicted membrane destabilizing activity of the polymer under acidic conditions typical of the endosomal compartment. Enhanced siRNA uptake was demonstrated in DoHH2 lymphoma and transduced HeLa-R cells expressing CD22 but not in CD22 negative HeLa-R cells. Gene knockdown was significantly improved with CD22-targeted vs. nontargeted polymeric micelles. Treatment of DoHH2 cells with CD22-targeted polymeric micelles containing 15 nmol/l siRNA produced 70% reduction of gene expression. This CD22-targeted polymer carrier may be useful for siRNA delivery to lymphoma cells. PMID- 21629224 TI - Stable human FIX expression after 0.9G intrauterine gene transfer of self complementary adeno-associated viral vector 5 and 8 in macaques. AB - Intrauterine gene transfer (IUGT) offers ontological advantages including immune naivete mediating tolerance to the vector and transgenic products, and effecting a cure before development of irreversible pathology. Despite proof-of-principle in rodent models, expression efficacy with a therapeutic transgene has yet to be demonstrated in a preclinical nonhuman primate (NHP) model. We aimed to determine the efficacy of human Factor IX (hFIX) expression after adeno-associated-viral (AAV)-mediated IUGT in NHP. We injected 1.0-1.95 * 10(13) vector genomes (vg)/kg of self-complementary (sc) AAV5 and 8 with a LP1-driven hFIX transgene intravenously in 0.9G late gestation NHP fetuses, leading to widespread transduction with liver tropism. Liver-specific hFIX expression was stably maintained between 8 and 112% of normal activity in injected offspring followed up for 2-22 months. AAV8 induced higher hFIX expression (P = 0.005) and milder immune response than AAV5. Random hepatocellular integration was found with no hotspots. Transplacental spread led to low-level maternal tissue transduction, without evidence of immunotoxicity or germline transduction in maternal oocytes. A single intravenous injection of scAAV-LP1-hFIXco to NHP fetuses in late gestation produced sustained clinically-relevant levels of hFIX with liver specific expression and a non-neutralizing immune response. These data are encouraging for conditions where gene transfer has the potential to avert perinatal death and long-term irreversible sequelae. PMID- 21629225 TI - A 10 patient case report on the impact of plasmapheresis upon neutralizing factors against adeno-associated virus (AAV) types 1, 2, 6, and 8. AB - Adeno-associated viruses (AAV) are small, nonenveloped single-stranded DNA viruses which require helper viruses to facilitate efficient replication. These recombinant viruses are some of the most promising candidates for therapeutic gene transfer to treat many genetic and acquired diseases. Nevertheless, the presence of humoral responses to the wild-type AAV common among humans is one of the limitations of in vivo transduction efficacy in humans using cognate recombinant vector. In this study, based on the serum samples that we were able to collect from various clinical situations, we studied the impact of one to five plasmapheresis (PP), at 1-5 day intervals on neutralizing factor (NAF) titers specific for AAV types 1, 2, 6, and 8 in seropositive patients with diverse pathologies and immunosuppressor treatments. We show that frequent sessions of PP result in drastic reduction of NAF specific for AAV1, 2, 6, and 8 to undetectable levels or titers <1:5, mainly when initial titers, i.e., before the first PP were <=1:20. Altogether, these results show that the use of PP and its possible association with pharmacological immunosuppressive treatments may help to design optimal management of seropositive patients for AAV gene therapy treatments. PMID- 21629226 TI - Synergistic inhibition of endochondral bone formation by silencing Hif1alpha and Runx2 in trauma-induced heterotopic ossification. AB - Angiogenesis and osteogenesis are tightly coupled during bone development. We studied the effect of inhibition of Hif1alpha and Runt-related protein 2 (Runx2) on the formation of heterotopic ossification (HO). We constructed lentivirus vectors expressing Hif1alpha small interfering RNA (siRNA) and Runx2 siRNA. The inhibition of Hif1alpha function impaired osteoblast proliferation while osteoblasts differentiated normally. Osteoblasts lacking Runx2 proliferated normally while the differentiation was impaired. The osteoblast differentiation was significantly inhibited by co-Runx2 and Hif1alpha siRNA treatment. The formation of HO by inhibiting Runx2 and Hif1alpha in an animal model induced by Achilles tenotomy was investigated. The results showed that lacking of Runx2 and Hif1alpha could inhibit HO formation. Inhibition of Hif1alpha prevented HO formation only at the initial step and inhibition of Runx2 worked both at the initial step and after chondrogenesis. Angiogenesis and the expressions of osteogenic genes were downregulated in the Hif1alpha siRNA group. We found synergistic inhibition of endochondral bone formation by silencing Hif1alpha and Runx2. Our study provided new insight into the roles of Hif1alpha and Runx2 during the processes of endochondral bone formation, and had important implications for the new therapeutic methods to inhibit HO or to enhance bone formation. PMID- 21629227 TI - Neural stem cell-based cell carriers enhance therapeutic efficacy of an oncolytic adenovirus in an orthotopic mouse model of human glioblastoma. AB - The potential utility of oncolytic adenoviruses as anticancer agents is significantly hampered by the inability of the currently available viral vectors to effectively target micrometastatic tumor burden. Neural stem cells (NSCs) have the ability to function as cell carriers for targeted delivery of an oncolytic adenovirus because of their inherent tumor-tropic migratory ability. We have previously reported that in vivo delivery of CRAd-S-pk7, a glioma-restricted oncolytic adenovirus, can enhance the survival of animals with experimental glioma. In this study, we show that intratumoral delivery of NSCs loaded with the CRAD-S-pk7 in an orthotopic xenograft model of human glioma is able to not only inhibit tumor growth but more importantly to increase median survival by ~50% versus animals treated with CRAd-S-pk7 alone (P = 0.0007). We also report that oncolytic virus infection upregulates different chemoattractant receptors and significantly enhances migratory capacity of NSCs both in vitro and in vivo. Our data further suggest that NSC-based carriers have the potential to improve the clinical efficacy of antiglioma virotherapy by not only protecting therapeutic virus from the host immune system, but also amplifying the therapeutic payload selectively at tumor sites. PMID- 21629231 TI - Pediatrics: Weighing the options: assessing fluid overload in children. PMID- 21629228 TI - Statins, inflammation and kidney disease. AB - Inflammation is highly prevalent in patients with chronic kidney disease (CKD) and is consistently associated with cardiovascular morbidity and mortality. Clinical event rates increase with declining renal function and activation of the acute-phase response. Statins are potent anti-inflammatory drugs that reduce the incidence of cardiovascular events. Owing to the increased prevalence of inflammation in patients with CKD and the potent effect of statins in individuals with elevated levels of C-reactive protein, these drugs should be especially effective in patients with CKD. Whereas data indicate that pravastatin may prevent loss of kidney function to a greater extent in individuals with evidence of increased inflammation than in those who show no inflammation, two large, randomized statin trials in patients on hemodialysis found no benefit of statin therapy, neither in the whole study group nor after stratifying for inflammation. Irrespective of inflammation, guidelines recommend treatment of dyslipidemia in early stages of CKD, which is supported by results from recent meta-analyses, and the Study of Heart and Renal Protection (SHARP), a large, randomized, placebo controlled trial. PMID- 21629229 TI - Diets and enteral supplements for improving outcomes in chronic kidney disease. AB - Protein-energy wasting (PEW), which is manifested by low serum levels of albumin or prealbumin, sarcopenia and weight loss, is one of the strongest predictors of mortality in patients with chronic kidney disease (CKD). Although PEW might be engendered by non-nutritional conditions, such as inflammation or other comorbidities, the question of causality does not refute the effectiveness of dietary interventions and nutritional support in improving outcomes in patients with CKD. The literature indicates that PEW can be mitigated or corrected with an appropriate diet and enteral nutritional support that targets dietary protein intake. In-center meals or oral supplements provided during dialysis therapy are feasible and inexpensive interventions that might improve survival and quality of life in patients with CKD. Dietary requirements and enteral nutritional support must also be considered in patients with CKD and diabetes mellitus, in patients undergoing peritoneal dialysis, renal transplant recipients, and in children with CKD. Adjunctive pharmacological therapies, such as appetite stimulants, anabolic hormones, and antioxidative or anti-inflammatory agents, might augment dietary interventions. Intraperitoneal or intradialytic parenteral nutrition should be considered for patients with PEW whenever enteral interventions are not possible or are ineffective. Controlled trials are needed to better assess the effectiveness of in-center meals and oral supplements. PMID- 21629232 TI - Calcium supplements and osteoporosis: the heart of the matter. PMID- 21629233 TI - Bone: Risedronate increases BMD in anorexia nervosa. PMID- 21629234 TI - Anomalous role for dietary salt in diabetes mellitus? PMID- 21629235 TI - Painful diabetic neuropathy--new choice of first-line therapy? PMID- 21629236 TI - Outcomes of diabetes mellitus in Asian Americans. PMID- 21629237 TI - Use of FRAX(r) to target BMD in women <65 years of age. PMID- 21629238 TI - Cystic fibrosis-related diabetes. PMID- 21629239 TI - The role of fecal elastase-1 in detecting exocrine pancreatic disease. AB - Exocrine pancreatic disease is thought to be uncommon in clinical practice and usually secondary to excess alcohol intake. Although excess alcohol intake does account for many cases of exocrine pancreatic disease, other conditions are associated with exocrine pancreatic insufficiency and such dysfunction perhaps occurs more frequently than conventionally expected. A reliable, patient friendly, cheap and easy to use test for exocrine pancreatic disease is yet to be established; however, in many countries the main (and often only available) method of assessment of exocrine pancreatic function is the fecal-elastase-1 test. This Review examines the role of fecal-elastase-1 testing in detecting exocrine pancreatic insufficiency in a number of gastrointestinal and nongastrointestinal conditions and determines the value of pancreatic enzyme supplementation in these settings. PMID- 21629240 TI - Genetic susceptibility to ischemic stroke. AB - Clinicians who treat patients with stroke need to be aware of several single-gene disorders that have ischemic stroke as a major feature, including sickle cell disease, Fabry disease, cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy, and retinal vasculopathy with cerebral leukodystrophy. The reported genome-wide association studies of ischemic stroke and several related phenotypes (for example, ischemic white matter disease) have shown that no single common genetic variant imparts major risk. Larger studies with samples numbering in the thousands are ongoing to identify common variants with smaller effects on risk. Pharmacogenomic studies have uncovered genetic determinants of response to warfarin, statins and clopidogrel. Despite increasing knowledge of stroke genetics, incorporating this new knowledge into clinical practice remains a challenge. The goals of this article are to review common single-gene disorders relevant to ischemic stroke, summarize the status of candidate gene and genome-wide studies aimed at discovering genetic stroke risk factors, and to briefly discuss pharmacogenomics related to stroke treatment. PMID- 21629242 TI - Experimental arthritis: Targeting joint lymphatic function. PMID- 21629241 TI - Epidemiology of CVD in rheumatic disease, with a focus on RA and SLE. AB - The excess risk of cardiovascular disease (CVD) associated with inflammatory rheumatic diseases has long been recognized. Patients with established rheumatoid arthritis (RA) or systemic lupus erythematosus (SLE) have higher mortality compared with the general population. Over 50% of premature deaths in RA are attributable to CVD. Excess mortality in SLE follows a bimodal pattern, with the early peak predominantly a consequence of active lupus or its complications, and the later peak largely attributable to atherosclerosis. Patients with RA or SLE are also at increased risk of nonfatal ischemic heart disease. The management and outcome of myocardial infarction and congestive heart failure in patients with RA or SLE differs from that in the general population. Traditional CVD risk factors (TRF) include increasing age, male gender, smoking, hypertension, hypercholesterolemia and diabetes. Whereas some TRFs are elevated in patients with RA or SLE, several are not, and others exhibit paradoxical relationships. Risk scores developed for the general population based on TRFs are likely, therefore, to underestimate CVD risk in RA and SLE. Until additional research and disease-specific risk prediction tools are available, current evidence supports aggressive treatment of disease activity, and careful screening for and management of TRFs. PMID- 21629243 TI - Rheumatoid arthritis: When should we use rituximab to treat RA? PMID- 21629244 TI - The prognostic influence of tumour-infiltrating lymphocytes in cancer: a systematic review with meta-analysis. AB - BACKGROUND: Tumour-infiltrating lymphocytes (TILs) are often found in tumours, presumably reflecting an immune response against the tumour. We carried out a systematic review and meta-analysis, aiming to establish pooled estimates for survival outcomes based on the presence of TILs in cancer. METHODS: A Pubmed and Embase literature search was designed. Studies were included, in which the prognostic significance of intratumoural CD3+, CD4+, CD8+, and FoxP3+ lymphocytes, as well as ratios between these subsets, were determined in solid tumours. RESULTS: In pooled analysis, CD3+ TILs had a positive effect on survival with a hazard ratio (HR) of 0.58 (95% confidence interval (CI) 0.43-0.78) for death, as did CD8+ TILs with a HR of 0.71 (95% CI 0.62-0.82). FoxP3+ regulatory TILs were not linked to overall survival, with a HR of 1.19 (95% CI 0.84-1.67). The CD8/FoxP3 ratio produced a more impressive HR (risk of death: HR 0.48, 95% CI 0.34-0.68), but was used in relatively few studies. Sample size and follow-up time seemed to influence study outcomes. CONCLUSION: Any future studies should be carefully designed, to prevent overestimating the effect of TILs on prognosis. In this context, ratios between TIL subsets may be more informative. PMID- 21629245 TI - Phase I dose-escalation study to determine the safety, pharmacokinetics and pharmacodynamics of brivanib alaninate in combination with full-dose cetuximab in patients with advanced gastrointestinal malignancies who have failed prior therapy. AB - BACKGROUND: The objectives of this phase I study were to determine the safety, pharmacokinetics (PK), pharmacodynamics and efficacy of brivanib combined with full-dose cetuximab in patients with advanced gastrointestinal malignancies. METHODS: Patients with advanced gastrointestinal malignancies who had failed prior therapies received brivanib (320, 600 or 800 mg daily) plus cetuximab (400 mg m(-2) loading dose then 250 mg m(-2) weekly). Assessments included adverse events, PK, tumour response, 2[18F]fluoro-2-deoxyglucose positron-emitting tomography and K-Ras mutation analyses. RESULTS: Toxicities observed were manageable; the most common treatment-related toxicities (>10% of patients) were fatigue, diarrhoea, anorexia, increase in aspartate aminotransferase and alanine aminotransferase, acneiform dermatitis, headache, mucosal inflammation, nausea, dry skin, vomiting, hypertension, pruritus, proteinuria and weight loss. Of 62 patients, 6 (9.7%) had objective radiographic partial responses, with an overall response rate of 10%. Median duration of response was 9.2 months; median progression-free survival was 3.9 months. CONCLUSIONS: The acceptable toxicity profile and efficacy of brivanib observed in this study were promising. These findings are being further evaluated in a phase III study of brivanib plus cetuximab vs cetuximab alone in patients previously treated with combination chemotherapy for K-Ras wild-type advanced metastatic colorectal cancer. PMID- 21629246 TI - Hepatitis B virus X protein downregulates expression of the miR-16 family in malignant hepatocytes in vitro. AB - BACKGROUND: Hepatitis B virus X protein (HBx) is involved in the initiation and progression of hepatocellular carcinoma (HCC) by regulating the host protein coding genes. In this study, we showed that HBx altered the expression of microRNAs (miRNAs) to promote proliferation and transformation in malignant hepatocytes in vitro. METHODS: miRNA microarray and quantitative reverse transcription polymerase chain reactions (qRT-PCRs) were performed to identify miRNAs that were differentially regulated by HBx in HCC cells. Protein, mRNA, and miRNA expression analyses; cell cycle and apoptosis analyses; loss/gain-of function analysis; and luciferase reporter assays were performed to delineate the consequences of miR-16 family repression in HepG2 cells. RESULTS: Hepatitis B virus X protein induced widespread deregulation of miRNAs in HepG2 cells, and the downregulation of the miR-16 family was reproducible in HepG2, SK-HEP-1, and Huh7 cells. CCND1, a target of the miR-16 family, was derepressed by HBx in HepG2 cells. c-Myc mediated the HBx-induced repression of miR-15a/16 in HepG2 cells. Ectopically expressed miR-15a/16 suppressed the proliferation, clonogenicity, and anchorage-independent growth of HBx-expressing HepG2 cells by arresting them in the G1 phase and inducing apoptosis, whereas reduced expression of miR-16 accelerated the growth and cell-cycle progression of HepG2 cells. CONCLUSIONS: Hepatitis B virus X protein altered the in vitro expression of miRNAs in host malignant hepatocytes, particularly downregulating the miR-16 family. Repression of miR-15a/16 is c-Myc mediated and is required for the HBx-induced transformation of HepG2 cells in vitro. Therefore, miR-16 family may serve as a therapeutic target for hepatitis B virus (HBV)-associated HCC. PMID- 21629247 TI - NF-kappaB mediates the 12(S)-HETE-induced endothelial to mesenchymal transition of lymphendothelial cells during the intravasation of breast carcinoma cells. AB - BACKGROUND: The intravasation of breast cancer into the lymphendothelium is an early step of metastasis. Little is known about the mechanisms of bulky cancer invasion into lymph ducts. METHODS: To particularly address this issue, we developed a 3-dimensional co-culture model involving MCF-7 breast cancer cell spheroids and telomerase-immortalised human lymphendothelial cell (LEC) monolayers, which resembles intravasation in vivo and correlated the malignant phenotype with specific protein expression of LECs. RESULTS: We show that tumour spheroids generate 'circular chemorepellent-induced defects' (CCID) in LEC monolayers through retraction of LECs, which was induced by 12(S) hydroxyeicosatetraenoic acid (HETE) secreted by MCF-7 spheroids. This 12(S)-HETE regulated retraction of LECs during intravasation particularly allowed us to investigate the key regulators involved in the motility and plasticity of LECs. In all, 12(S)-HETE induced pro-metastatic protein expression patterns and showed NF-kappaB-dependent up-regulation of the mesenchymal marker protein S100A4 and of transcriptional repressor ZEB1 concomittant with down-regulation of the endothelial adherence junction component VE-cadherin. This was in accordance with ~50% attenuation of CCID formation by treatment of cells with 10 MUM Bay11-7082. Notably, 12(S)-HETE-induced VE-cadherin repression was regulated by either NF kappaB or by ZEB1 since ZEB1 siRNA knockdown abrogated not only 12(S)-HETE mediated VE-cadherin repression but inhibited VE-cadherin expression in general. INTERPRETATION: These data suggest an endothelial to mesenchymal transition-like process of LECs, which induces single cell motility during endothelial transmigration of breast carcinoma cells. In conclusion, this study demonstrates that the 12(S)-HETE-induced intravasation of MCF-7 spheroids through LECs require an NF-kappaB-dependent process of LECs triggering the disintegration of cell-cell contacts, migration, and the generation of CCID. PMID- 21629248 TI - Coffee intake and oral-oesophageal cancer: follow-up of 389,624 Norwegian men and women 40-45 years. AB - BACKGROUND: The evidence on the relationship between coffee intake and cancer of the oral cavity and oesophagus is conflicting and few follow-up studies have been done. METHODS: A total of 389,624 men and women 40-45 years who participated in a national survey programme were followed with respect to cancer for an average of 14.4 years by linkage to the Cancer Registry of Norway. Coffee consumption at baseline was reported as a categorical variable (0 or <1 cup, 1-4, 5-8, 9+ cups per day). RESULTS: Altogether 450 squamous oral or oesophageal cancers were registered during follow-up. The adjusted hazard ratios with 1-4 cups per day as reference were 1.01 (95% confidence interval: 0.70, 1.47), 1.16 (0.93, 1.45) and 0.96 (0.71, 1.14) for 0 or <1 cup, 5-8 and 9+ cups per day, respectively. Stratification by sex, type of coffee, smoking status and dividing the end point into oral and oesophageal cancers gave heterogeneous and non-significant estimates. CONCLUSION: This study does not support an inverse relationship between coffee intake and incidence of cancer in the mouth or oesophagus, but cannot exclude a weak inverse relationship. PMID- 21629249 TI - End-of-study safety, immunogenicity, and efficacy of quadrivalent HPV (types 6, 11, 16, 18) recombinant vaccine in adult women 24-45 years of age. AB - BACKGROUND: Previous analyses from a randomised trial in women aged 24-45 years have shown the quadrivalent human papillomavirus (qHPV) vaccine to be efficacious in the prevention of infection, cervical intraepithelial neoplasia (CIN), and external genital lesions (EGLs) related to HPV 6/11/16/18. In this report, we present end-of-study efficacy, safety, and immunogenicity data with a median follow-up time of 4.0 years. METHODS: We enrolled 3819 24-45-year-old women with no history of cervical disease or genital warts in the past 5 years. Women received quadrivalent vaccine or placebo at day 1, and at months 2 and 6. Ascertainment of CIN/EGL was accomplished through Pap testing, genital inspection, and cervicovaginal sampling (every 6 months). The main analysis was conducted in a per-protocol efficacy population (that received three doses, was naive to the relevant HPV types at day 1, and remained free of infection through month 7). Efficacy was also estimated in other naive and non-naive populations. RESULTS: Vaccine efficacy against the combined incidence of persistent infection, CIN/EGL related to HPV6/11/16/18 in the per-protocol population was 88.7% (95% CI: 78.1, 94.8). Efficacy for women who were seropositive and DNA negative for the relevant vaccine HPV type at the time of enrolment who received at least 1 dose was 66.9% (95% CI: 4.3, 90.6). At month 48, 91.5, 92.0, 97.4, and 47.9% of vaccinated women were seropositive to HPV 6/11/16/18, respectively. No serious vaccine-related adverse experiences were reported. CONCLUSIONS: The qHPV vaccine demonstrated high efficacy, immunogenicity, and acceptable safety in women aged 24-45 years, regardless of previous exposure to HPV vaccine type. PMID- 21629254 TI - Is RNAi dead? PMID- 21629257 TI - AAV9: over the fence and into the woods . . . PMID- 21629258 TI - Cerebral glucose utilisation in hepatitis C virus infection-associated encephalopathy. AB - Patients with hepatitis C virus (HCV) infection frequently show neuropsychiatric symptoms. This study aims to help clarify the neurochemical mechanisms behind these symptoms and to add further proof to the hypothesis that HCV may affect brain function. Therefore, 15 patients who reported increasing chronic fatigue, mood alterations, and/or cognitive decline since their HCV infection underwent neurologic and neuropsychological examination, magnetic resonance imaging, (18)F fluoro-deoxy-glucose positron emission tomography of the brain, and single photon emission tomography of striatal dopamine and midbrain serotonin transporter (SERT) availability. None of the patients had liver cirrhosis. Patients' data were compared with data of age-matched controls. In addition, regression analysis was performed between cognitive deficits, and mood and fatigue scores as dependent variables, and cerebral glucose metabolism, dopamine, or SERT availability as predictors. Patients showed significant cognitive deficits, significantly decreased striatal dopamine and midbrain SERT availability, and significantly reduced glucose metabolism in the limbic association cortex, and in the frontal, parietal, and superior temporal cortices, all of which correlated with dopamine transporter availability and psychometric results. Thus, the study provides further evidence of central nervous system affection in HCV-afflicted patients with neuropsychiatric symptoms. Data indicate alteration of dopaminergic neurotransmission as a possible mechanism of cognitive decline. PMID- 21629259 TI - A computational simulation of the effect of hemodilution on oxygen transport in middle cerebral artery vasospasm. AB - Cerebral vasospasm after aneurysmal subarachnoid hemorrhage is a potentially severe sequel. The induction of hypertension, hypervolemia, and hemodilution is advocated for vasospasm, but it is unclear whether hemodilution confers any benefit. A finite element model of oxygen transport in the proximal middle cerebral artery (MCA) was used to evaluate the complex relationship among hematocrit, viscosity, oxygen content, and blood flow in the setting of vasospasm. A single-phase non-Newtonian finite element model based on three dimensional incompressible Navier-Stokes equations was constructed of the M1 segment. The model was solved at vessel stenoses ranging from 0% to 90% and hematocrit from 0.2 to 0.6. A small area of poststenotic recirculation was seen with mild (30%) stenosis. Poststenotic eddy formation was noted with more severe (60% to 90%) stenosis. Volumetric flow was inversely related to hematocrit at mild stenosis (0% to 30%). With near-complete stenosis (90%), a paradoxical increase in flow was seen with increasing hematocrit. Oxygen transport across the segment was related to hematocrit at all levels of stenosis with increasing oxygen transport despite a reduction in blood flow, suggesting that with clinically significant vasospasm in the MCA, hemodilution does not improve oxygen transport, but to the contrary, that ischemia may be worsened. PMID- 21629260 TI - Validating a local Arterial Input Function method for improved perfusion quantification in stroke. AB - In bolus-tracking perfusion magnetic resonance imaging (MRI), temporal dispersion of the contrast bolus due to stenosis or collateral supply presents a significant problem for accurate perfusion quantification in stroke. One means to reduce the associated perfusion errors is to deconvolve the bolus concentration time-course data with local Arterial Input Functions (AIFs) measured close to the capillary bed and downstream of the arterial abnormalities causing dispersion. Because the MRI voxel resolution precludes direct local AIF measurements, they must be extrapolated from the surrounding data. To date, there have been no published studies directly validating these local AIFs. We assess the effectiveness of local AIFs in reducing dispersion-induced perfusion error by measuring the residual dispersion remaining in the local AIF deconvolved perfusion maps. Two approaches to locating the local AIF voxels are assessed and compared with a global AIF deconvolution across 19 bolus-tracking data sets from patients with stroke. The local AIF methods reduced dispersion in the majority of data sets, suggesting more accurate perfusion quantification. Importantly, the validation inherently identifies potential areas for perfusion underestimation. This is valuable information for the identification of at-risk tissue and management of stroke patients. PMID- 21629262 TI - Potential for interdependent development of tRNA determinants for aminoacylation and ribosome decoding. AB - Although the nucleotides in tRNA required for aminoacylation are conserved in evolution, bacterial aminoacyl-transfer RNA synthetases are unable to acylate eukaryotic tRNA. The cross-species barrier may be due to the absence of eukaryote specific domains from bacterial aminoacyl-transfer RNA synthetases. Here we show that whereas Escherichia coli CysRS cannot acylate human tRNA(Cys), the fusion of a eukaryote-specific domain of human CysRS overcomes the cross-species barrier in human tRNA(Cys). In addition to enabling recognition of the sequence differences in the tertiary core of tRNA(Cys), the fused eukaryotic domain redirects the specificity of E. coli CysRS from the A37 present in bacterial tRNA(Cys) to the G37 in mammals. Further experiments show that the accuracy of codon recognition on the ribosome was also highly sensitive to the A37G transition in tRNA(Cys). These results raise the possibility of the development of tRNA nucleotide determinants for aminoacylation being interdependent with those for ribosome decoding. PMID- 21629263 TI - TRAF6 ubiquitinates TGFbeta type I receptor to promote its cleavage and nuclear translocation in cancer. AB - Transforming growth factor beta (TGFbeta) is a pluripotent cytokine promoting epithelial cell plasticity during morphogenesis and tumour progression. TGFbeta binding to type II and type I serine/threonine kinase receptors (TbetaRII and TbetaRI) causes activation of different intracellular signaling pathways. TbetaRI is associated with the ubiquitin ligase tumor necrosis factor receptor (TNFR) associated factor 6 (TRAF6). Here we show that TGFbeta, via TRAF6, causes Lys63 linked polyubiquitination of TbetaRI, promoting cleavage of TbetaRI by TNF-alpha converting enzyme (TACE), in a PKCzeta-dependent manner. The liberated intracellular domain (ICD) of TbetaRI associates with the transcriptional regulator p300 to activate genes involved in tumour cell invasiveness, such as Snail and MMP2. Moreover, TGFbeta-induced invasion of cancer cells is TACE- and PKCzeta- dependent and the TbetaRI ICD is localized in the nuclei of different kinds of tumour cells in tissue sections. Thus, our data reveal a specific role for TbetaRI in TGFbeta mediated tumour invasion. PMID- 21629265 TI - Contemporary and historical separation of transequatorial migration between genetically distinct seabird populations. AB - Pelagic seabirds are highly mobile, reducing the likelihood of allopatric speciation where disruption of gene flow between populations is caused by physically insurmountable, extrinsic barriers. Spatial segregation during the non breeding season appears to provide an intrinsic barrier to gene flow among seabird populations that otherwise occupy nearby or overlapping regions during breeding, but how this is achieved remains unclear. Here we show that the two genetically distinct populations of Cook's petrel (Pterodroma cookii) exhibit transequatorial separation of non-breeding ranges at contemporary (ca. 2-3 yrs) and historical (ca. 100 yrs) time scales. Segregation during the non-breeding season per se appears as an unlikely barrier to gene flow. Instead we provide evidence that habitat specialization during the non-breeding season is associated with breeding asynchrony which, in conjunction with philopatry, restricts gene flow. Habitat specialization during breeding and non-breeding likely promotes evolutionary divergence between these two populations via local adaptation. PMID- 21629267 TI - Genetics and the environment converge to dysregulate N-glycosylation in multiple sclerosis. AB - How environmental factors combine with genetic risk at the molecular level to promote complex trait diseases such as multiple sclerosis (MS) is largely unknown. In mice, N-glycan branching by the Golgi enzymes Mgat1 and/or Mgat5 prevents T cell hyperactivity, cytotoxic T-lymphocyte antigen 4 (CTLA-4) endocytosis, spontaneous inflammatory demyelination and neurodegeneration, the latter pathologies characteristic of MS. Here we show that MS risk modulators converge to alter N-glycosylation and/or CTLA-4 surface retention conditional on metabolism and vitamin D(3), including genetic variants in interleukin-7 receptor alpha (IL7RA*C), interleukin-2 receptor-alpha (IL2RA*T), MGAT1 (IV(A)V(T-T)) and CTLA-4 (Thr17Ala). Downregulation of Mgat1 by IL7RA*C and IL2RA*T is opposed by MGAT1 (IV(A)V(T-T)) and vitamin D(3), optimizing branching and mitigating MS risk when combined with enhanced CTLA-4 N-glycosylation by CTLA-4 Thr17. Our data suggest a molecular mechanism in MS whereby multiple environmental and genetic inputs lead to dysregulation of a final common pathway, namely N-glycosylation. PMID- 21629268 TI - Folate and related micronutrients, folate-metabolising genes and risk of ovarian cancer. AB - BACKGROUND/OBJECTIVE: Folates are essential for DNA synthesis and methylation, and thus may have a role in carcinogenesis. Limited evidence suggests folate containing foods might protect against some cancers and may partially mitigate the increased risk of breast cancer associated with alcohol intake, but there is little information regarding ovarian cancer. Our aim was to evaluate the role of folate and related micronutrients, polymorphisms in key folate-metabolising genes and environmental factors in ovarian carcinogenesis. SUBJECTS/METHODS: Participants in the Australian Ovarian Cancer Study (1363 cases, 1414 controls) self-completed risk factor and food-frequency questionnaires. DNA samples (1638 cases, 1278 controls) were genotyped for 49 tag single-nucleotide polymorphisms (SNPs) in the methylene tetrahydrofolate reductase (MTHFR), methionine synthase (MTR) and MTR reductase (MTRR) genes. Logistic regression models were used to generate adjusted odds ratios and 95% confidence intervals. RESULTS: We saw no overall association between the intake of folate, B vitamins or other methyl donors and ovarian cancer risk, although increasing folate from foods was associated with reduced risk among current smokers (P(trend)=0.03) and folic acid intake was associated with borderline significant increased risks among women who consumed >=1 standard alcoholic drinks/day (odds ratio (OR)=1.64; 95% confidence interval (CI) 1.05-2.54, P(trend)=0.05). Two SNPs (rs7365052, rs7526063) showed borderline significant inverse associations with ovarian cancer risk; both had very low minor allele frequencies. There was little evidence for interaction between genotype and micronutrient intake or for variation between different histological subtypes of ovarian cancer. CONCLUSIONS: Our data provide little evidence to support a protective role for folate in ovarian carcinogenesis but suggest further evaluation of the joint effects of folic acid and alcohol is warranted. PMID- 21629269 TI - Habitual dietary intake, eating pattern and physical activity of women with polycystic ovary syndrome. AB - BACKGROUND/OBJECTIVE: Diet and lifestyle modifications may be of benefit in the management of polycystic ovary syndrome (PCOS), but there is a paucity of data on these behaviours in women with PCOS. This study aims to address this through a comprehensive investigation of the habitual diet and activity patterns of UK women with PCOS and their alignment with dietary recommendations for health. SUBJECTS/METHODS: A 7-day-estimated food and activity diary and questionnaire was completed by 210 women with PCOS for calculation of activity levels, energy and nutrient intakes and dietary glycaemic index (GI). RESULTS: Mean (s.d.) body mass index (BMI) was 27.4 (7.3) kg/m(2) (n=183), with 53% of women with PCOS having a BMI>25 kg/m(2). Of the overweight women, approximately half were not achieving sufficient physical activity to promote weight loss. More frequent eating episodes and a lower BMI were weakly associated (r=-0.158, P=0.034). Mean percentage energy from fat was 38 (7)% (12% energy from saturated fat), with 68% of women with PCOS consuming > 35% energy from fat. Mean dietary GI was higher in obese women with PCOS, compared with healthy weight women with PCOS (55.7 (3.4) and 53.8 (4.0), respectively; P=0.043). CONCLUSION: Many women with PCOS are not achieving dietary intakes and levels of physical activity that optimise symptom management and disease prevention. Advice should focus on fat quality and quantity and carbohydrate modification. There is a need for further robust research into the role of dietary GI in the PCOS population. PMID- 21629270 TI - Effect of camel milk on glycemic control and insulin requirement in patients with type 1 diabetes: 2-years randomized controlled trial. AB - BACKGROUND/OBJECTIVES: Hypoglycemic effect of camel milk supplementation in experimental rat model and significant reduction in doses of insulin in type 1 diabetic patients have been observed in our previous studies. This long-term study was undertaken to assess the efficacy, safety and acceptability of camel milk as an adjunct to insulin therapy in type 1 diabetics. SUBJECTS/METHODS: In this 2-year randomized clinical, parallel design study, 24 type 1 diabetics were enrolled and divided into two groups. Group I (n=12) received usual care, that is, diet, exercise and insulin and Group II (n=12) received 500 ml camel milk in addition to the usual care. Insulin requirement was titrated weekly by blood glucose estimation. Results were analyzed by using the regression technique. RESULTS: In camel milk group, there was decrease in mean blood glucose (118.58+/ 19-93.16+/-17.06 mg/dl), hemoglobin A1c levels (7.81+/-1.39-5.44+/-0.81%) and insulin doses (32.50+/-9.99-17.50+/-12.09 U/day, P<0.05). Out of 12 subjects receiving camel milk, insulin requirement in 3 subjects reduced to zero. There was nonsignificant change in plasma insulin and anti-insulin antibodies in both the groups. CONCLUSION: It may be stated that camel milk is safe and efficacious in improving long-term glycemic control, with a significant reduction in the doses of insulin in type 1 diabetic patients. PMID- 21629271 TI - Mitochondria breathe for autophagy. PMID- 21629272 TI - Arp you ready for actin in the nucleus? PMID- 21629273 TI - P53 conformational switching for selectivity may reveal a general solution for specific DNA binding. PMID- 21629275 TI - The future of direct-to-consumer clinical genetic tests. AB - In light of the meeting of the US Food and Drug Administration (FDA) in March 2011 to discuss the regulation of clinical direct-to-consumer (DTC) genetic tests, we have invited five experts to consider the best means of overseeing the ordering and interpretation of these tests. Should these tests be regulated? If so, who, if anyone, should communicate results to consumers? PMID- 21629274 TI - Family-based designs for genome-wide association studies. AB - Association mapping has successfully identified common SNPs associated with many diseases. However, the inability of this class of variation to account for most of the supposed heritability has led to a renewed interest in methods - primarily linkage analysis - to detect rare variants. Family designs allow for control of population stratification, investigations of questions such as parent-of-origin effects and other applications that are imperfectly or not readily addressed in case-control association studies. This article guides readers through the interface between linkage and association analysis, reviews the new methodologies and provides useful guidelines for applications. Just as effective SNP-genotyping tools helped to realize the potential of association studies, next-generation sequencing tools will benefit genetic studies by improving the power of family based approaches. PMID- 21629277 TI - Mining for therapeutic gold. AB - A comprehensive and collaborative strategy to enable the investigation of new uses of approved and abandoned drug compounds could advance translational research. PMID- 21629278 TI - Could pharma open its drug freezers? PMID- 21629279 TI - The long march of antisense. PMID- 21629281 TI - Trial watch: Phase III success for Biogen's oral multiple sclerosis therapy. PMID- 21629282 TI - Deal watch: is deal-making becoming more difficult? PMID- 21629283 TI - Court refuses to hurry generic Lipitor decision. PMID- 21629284 TI - Richard Bergstrom. Interview by Asher Mullard. PMID- 21629285 TI - US academic drug discovery. PMID- 21629286 TI - Ipilimumab. PMID- 21629288 TI - Anticancer drugs: Now get out of that.... PMID- 21629287 TI - Cancer: Lipoxygenase makes a leaky tumour. PMID- 21629289 TI - Mood disorders: Small-molecule neurotrophin antagonist reduces anxiety. PMID- 21629291 TI - Sepsis: DAMPening inflammation. PMID- 21629293 TI - The productivity crisis in pharmaceutical R&D. AB - Advances in the understanding of the molecular basis of diseases have expanded the number of plausible therapeutic targets for the development of innovative agents in recent decades. However, although investment in pharmaceutical research and development (R&D) has increased substantially in this time, the lack of a corresponding increase in the output in terms of new drugs being approved indicates that therapeutic innovation has become more challenging. Here, using a large database that contains information on R&D projects for more than 28,000 compounds investigated since 1990, we examine the decline of R&D productivity in pharmaceuticals in the past two decades and its determinants. We show that this decline is associated with an increasing concentration of R&D investments in areas in which the risk of failure is high, which correspond to unmet therapeutic needs and unexploited biological mechanisms. We also investigate the potential variations in productivity with regard to the regional location of companies and find that although companies based in the United States and Europe differ in the composition of their R&D portfolios, there is no evidence of any productivity gap. PMID- 21629292 TI - Principles and mechanisms of vessel normalization for cancer and other angiogenic diseases. AB - Despite having an abundant number of vessels, tumours are usually hypoxic and nutrient-deprived because their vessels malfunction. Such abnormal milieu can fuel disease progression and resistance to treatment. Traditional anti angiogenesis strategies attempt to reduce the tumour vascular supply, but their success is restricted by insufficient efficacy or development of resistance. Preclinical and initial clinical evidence reveal that normalization of the vascular abnormalities is emerging as a complementary therapeutic paradigm for cancer and other vascular disorders, which affect more than half a billion people worldwide. Here, we discuss the mechanisms, benefits, limitations and possible clinical translation of vessel normalization for cancer and other angiogenic disorders. PMID- 21629294 TI - Immunomodulatory therapy to preserve pancreatic beta-cell function in type 1 diabetes. AB - Type 1 diabetes is a common, severe chronic autoimmune disease that is characterized by the progressive and insidious loss of self-tolerance to the insulin-producing pancreatic islet beta-cells. This loss of self-tolerance leads to the destruction of beta-cells and the development of overt hyperglycaemia at diagnosis. The incidence and prevalence of type 1 diabetes is rapidly increasing worldwide, and this has led to intensive efforts to develop immunotherapies to induce remission of the disease and improve clinical outcomes. Immunotherapy aims to restore self-tolerance, resulting in the downregulation of autoimmune responses to pancreatic self-antigens and arrested ongoing beta-cell destruction. When combined with replacement of the lost insulin-producing cells, this may lead to the restoration of euglycaemia. In this review, we discuss the current knowledge of the immunopathogenesis of type 1 diabetes and how this information has been translated into clinical trials. We also discuss next-generation combination immunotherapies that may be administered as adjuvant therapy at time of diagnosis. PMID- 21629296 TI - Rapid aneuploidy detection or karyotyping? Ethical reflection. AB - No consensus exists whether women at increased risk for trisomy 21, 13, and 18 should be offered stand-alone rapid aneuploidy detection (RAD) or karyotyping. In this paper, the ethical implications of a fast, relatively cheap and targeted RAD are examined. The advantages of RAD seem less robust than its proponents suggest. Fast test results only give a short-term psychological benefit. The cost advantage of RAD is apparent, but must be weighed against consequences like missed abnormalities, which are evaluated differently by professionals and pregnant women. Since pre-test information about RAD will have to include telling women about karyotyping as a possible alternative, the advantage of RAD in terms of the quantity of information that needs to be given may also be smaller than suggested. We conclude that none of the supposed arguments in favour of RAD is decisive in itself. Whether the case for RAD may still be regarded as convincing when taking these arguments together seems to depend on one's implicit view of what prenatal screening is about. Are we basically dealing with a test for trisomy 21 and a few conditions more? Or are there good grounds for also testing for the wider range of abnormalities that karyotyping can detect? As professionals and pregnant women may have different views about this, we suggest that the best approach is to offer women a choice between RAD and karyotyping. This approach is most in line with the general aim of prenatal screening: providing opportunities for autonomous reproductive choice. PMID- 21629297 TI - Are the kids really all right? Direct-to-consumer genetic testing in children: are company policies clashing with professional norms? AB - The genetic testing of minors within the direct-to-consumer (DTC) genetic testing (GT) context has been given relatively little attention. The issue of testing healthy children for diseases that would only develop in adulthood raises many important ethical, legal and social issues. As genetic testing is now available outside of the traditional health care system, often without even the intermediate of a health care professional, we surveyed 37 DTC GT companies regarding their policies for testing in children. Although the response rate is relatively low (35%, 13/37), our findings reveal that a clear majority of companies do perform genetic testing in minors. As such, companies testing for adult onset diseases are acting in contradiction of established professional guidelines, which state, among others, that, for predictive genetic testing, the availability of therapeutic or preventive measures is necessary for testing to be performed in asymptomatic minors. The community of stakeholders in children's health care and genetic testing should, therefore, decide which standards need to be upheld by DTC GT companies and ensure that these are met. PMID- 21629295 TI - Combating oxidative stress in vascular disease: NADPH oxidases as therapeutic targets. AB - NADPH oxidases are a family of enzymes that generate reactive oxygen species (ROS). The NOX1 (NADPH oxidase 1) and NOX2 oxidases are the major sources of ROS in the artery wall in conditions such as hypertension, hypercholesterolaemia, diabetes and ageing, and so they are important contributors to the oxidative stress, endothelial dysfunction and vascular inflammation that underlies arterial remodelling and atherogenesis. In this Review, we advance the concept that compared to the use of conventional antioxidants, inhibiting NOX1 and NOX2 oxidases is a superior approach for combating oxidative stress. We briefly describe some common and emerging putative NADPH oxidase inhibitors. In addition, we highlight the crucial role of the NADPH oxidase regulatory subunit, p47phox, in the activity of vascular NOX1 and NOX2 oxidases, and suggest how a better understanding of its specific molecular interactions may enable the development of novel isoform-selective drugs to prevent or treat cardiovascular diseases. PMID- 21629298 TI - Homozygosity mapping in 64 Syrian consanguineous families with non-specific intellectual disability reveals 11 novel loci and high heterogeneity. AB - Non-specific intellectual disability of autosomal recessive inheritance (NS-ARID) represents an important fraction of severe cognitive dysfunction disorders. To date, only 10 genes have been identified, and further 24 linked-ARID loci have been reported, as well as others with suggestive linkage. To discover novel genes causing NS-ARID, we undertook genome-wide homozygosity mapping in 64 consanguineous multiplex families of Syrian descent. A total of 11 families revealed unique, significantly linked loci at 4q26-4q28 (MRT17), 6q12-q15 (MRT18), 18p11 (MRT19), 16p12-q12 (MRT20), 11p15 (MRT21), 11p13-q14 (MRT23), 6p12 (MRT24), 12q13-q15 (MRT25), 14q11-q12 (MRT26), 15q23-q26 (MRT27), and 6q26-q27 (MRT28), respectively. Loci ranged between 1.2 and 45.6 Mb in length. One family showed linkage to chromosome 8q24.3, and we identified a mutation in TRAPPC9. Our study further highlights the extreme heterogeneity of NS-ARID, and suggests that no major disease gene is to be expected, at least in this study group. Systematic analysis of large numbers of affected families, as presented here, will help discovering the genetic causes of ID. PMID- 21629299 TI - Fatty acid desaturase 2 promoter mutation is not responsible for Delta6 desaturase deficiency. AB - Dietary essential polyunsaturated fatty acids (PUFAs) require fatty acid desaturases (FADS) for conversion to long-chain PUFAs (LCPUFAs), which are critical for many aspects of human health. A Delta6-desaturase deficiency in a single patient was attributed to an insertion mutation in the FADS2 promoter. Later population studies have shown this thymidine nucleotide (T) insertion to be a common polymorphism (rs3834458). We examined correlations between rs3834458 variants and fatty acid evidence of FADS2 activity in a cohort of rheumatoid arthritis patients selected for low or nil consumption of n-3 LCPUFA as fish or fish oil. The presence of the T allele was associated with higher FADS2 activity, as indicated by higher conversion of plasma n-3 PUFA to LCPUFA. However, the T insertion/deletion polymorphism did not affect FADS2 promoter activity in luciferase reporter assays in HepG2 or NIH/3T3 cells. Our results indicate that the polymorphism rs3834458 does not appear to directly affect FADS2 promoter activity and is not responsible for a previously reported Delta6-desaturase deficiency. PMID- 21629300 TI - 17p13.3 microduplications are associated with split-hand/foot malformation and long-bone deficiency (SHFLD). AB - Split-hand/foot malformation with long-bone deficiency (SHFLD) is a relatively rare autosomal-dominant skeletal disorder, characterized by variable expressivity and incomplete penetrance. Although several chromosomal loci for SHFLD have been identified, the molecular basis and pathogenesis of most SHFLD cases are unknown. In this study we describe three unrelated kindreds, in which SHFLD segregated with distinct but overlapping duplications in 17p13.3, a region previously linked to SHFLD. In a large three-generation family, the disorder was found to segregate with a 254 kb microduplication; a second microduplication of 527 kb was identified in an affected female and her unaffected mother, and a 430 kb microduplication versus microtriplication was identified in three affected members of a multi-generational family. These findings, along with previously published data, suggest that one locus responsible for this form of SHFLD is located within a 173 kb overlapping critical region, and that the copy gains are incompletely penetrant. PMID- 21629301 TI - Phenotypic heterogeneity in British patients with a founder mutation in the FHL1 gene. AB - Mutations in the four-and-a-half LIM domain 1 (FHL1) gene, which encodes a 280 amino-acid protein containing four LIM domains and a single zinc-finger domain in the N-terminal region, have been associated with a broad clinical spectrum of X linked muscle diseases encompassing a variety of different phenotypes. Patients might present with a scapuloperoneal myopathy, a myopathy with postural muscle atrophy and generalized hypertrophy, an Emery-Dreifuss muscular dystrophy, or an early onset myopathy with reducing bodies. It has been proposed that the phenotypic variability is related to the position of the mutation within the FHL1 gene. Here, we report on three British families with a heterogeneous clinical presentation segregating a single FHL1 gene mutation and haplotype, suggesting that this represents a founder mutation. The underlying FHL1 gene mutation was detected by direct sequencing and the founder effect was verified by haplotype analysis of the FHL1 gene locus. A 3-bp insertion mutation (p.Phe127_Thr128insIle) within the second LIM domain of the FHL1 gene was identified in all available affected family members of the three families. Haplotype analysis of the FHL1 region on Xq26 revealed that the families shared a common haplotype. The p.Phe127_Thr128insIle mutation in the FHL1 gene therefore appears to be a British founder mutation and FHL1 gene screening, in particular of exon 6, should therefore be indicated in British patients with a broad phenotypic spectrum of X-linked muscle diseases. PMID- 21629302 TI - From and to a very grey area. PMID- 21629303 TI - Mitochondrial transformation: time for concerted action. PMID- 21629304 TI - Complexity: The ultimate frontier? PMID- 21629305 TI - Comment on 'Jobs for the boys'. PMID- 21629306 TI - Teaming up for biomarker future. Many problems still hinder the use of biomarkers in clinical practice, but new public-private partnerships could improve the situation. PMID- 21629307 TI - A new era for plant biotechnology. A combination of pharmaceutical applications, technologies and safety measures could lead to new applications for genetically modified plants. PMID- 21629308 TI - Lightpipe device for delivery of uniform illumination for photodynamic therapy of the oral cavity. AB - A compact and efficient lightpipe device to deliver light to the human oral cavity for photodynamic therapy was designed and fabricated, having dimensions 6.8 mm * 6.8 mm * 46 mm. An average irradiance of 76 mW/cm2 with an average deviation of 5% was measured on a square 25 mm2 treatment field for an input power of 100 mW. The device limits irradiation of healthy tissue and offers potential for improvement over the current treatment procedure, which requires shielding of the whole cavity to avoid damage to healthy tissue. PMID- 21629309 TI - Gradient polymer-disposed liquid crystal single layer of large nematic droplets for modulation of laser light. AB - The light modulating ability of gradient polymer-disposed liquid crystal (PDLC) single layer of large droplets formed by nematic E7 in UV-cured polymer NOA65 is studied. Operating at relatively low voltages, such PDLC film with a of thickness 10-25 MUm and droplet size up to 50 MUm exhibits a good contrast ratio and is capable of producing a large phase shift for the propagating coherent light. For a linearly polarized He-Ne laser (lambda=633 nm), an electrically commanded phase shift as large as pi/2 can be obtained by the large-droplet region of the film. The electrically produced phase shift and its spatial profile controlled by the thickness of the gradient PDLC single layers of large nematic droplets can be useful for tunable spatial light modulators and other devices for active control of laser light. PMID- 21629310 TI - Dual-fiber collimator with elliptical spot for optical MEMS devices. AB - Fiber collimators are widely used in optical communication components and fiber optic sensors. Ordinary fiber collimators are made with a circular beam waist radius from 100 MUm to 300 MUm. The circular beam waist is too large to switch or shut the beam for certain micro-electro-mechanical system (MEMS) actuators (such as MEMS linear mirrors). In this paper, a dual-fiber collimator with an elliptical spot is proposed to meet the demands of MEMS optical devices. The elliptical spot collimator has been designed and fabricated, the beam waist spot of which is an elliptical spot with a 231.6 MUm long-axis radius and a 12.87 MUm short-axis radius, and its coupling loss is 0.37 dB. PMID- 21629311 TI - Point diffraction interferometer with adjustable fringe contrast for testing spherical surfaces. AB - A point diffraction interferometer (PDI) with adjustable fringe contrast is presented for the high-precision testing of spherical surfaces. The polarizing components are employed in the PDI to transform the polarization states of the test and reference beams, and a good fringe contrast can be realized by adjusting the relative intensities of interfering waves. The proposed system is compact and simple in structure, and it provides a feasible way for high-precision testing of spherical surfaces with low reflectivity. The theory of the interferometer is introduced in detail, along with the properties of optical components employed in the system, numerical analysis of systematic error, and the corresponding calibration procedure. Compared with the testing results of the ZYGO interferometer, a high accuracy with RMS value about 0.0025lambda is achieved with the proposed interferometer. Finally, the error consideration in the experiment is discussed. PMID- 21629312 TI - Self-organized 2D periodic arrays of nanostructures in silicon by nanosecond laser irradiation. AB - We report a phenomenon of spontaneous formation of self-organized 2D periodic arrays of nanostructures (protrusions) by directly exposing a silicon surface to multiple nanosecond laser pulses. These self-organized 2D periodic nanostructures are produced toward the edge as an annular region around the circular laser spot. The heights of these nanostructures are around 500 nm with tip diameter ~100 nm. The period of the nanostructures is about 1064 nm, the wavelength of the incident radiation. In the central region of the laser spot, nanostructures are destroyed because of the higher laser intensity (due to the Gaussian shape of the laser beam) and accumulation of large number of laser pulses. Optical diffraction from these nanostructures indicates a threefold symmetry, which is in accordance with the observed morphological symmetries of these nanostructures. PMID- 21629313 TI - Application of "parallel" moire deflectometry and the single beam Z-scan technique in the measurement of the nonlinear refractive index. AB - In this paper, the application of "parallel" moire deflectometry in measuring the nonlinear refractive index of materials is reported. In "parallel" moire deflectometry the grating vectors are parallel, and the resulting moire fringes are also parallel to the grating lines. Compared to "rotational" moire deflectometry and the Z-scan technique, which cannot easily determine the moire fringe's angle of rotation and is sensitive to power fluctuations, respectively, "parallel" moire deflectometry is more reliable, which allows one to measure the radius of curvature of the light beam by measuring the moire fringe spacing. The nonlinear refractive index of the sample, including the sense of the change, is obtained from the moire fringe spacing curve. The method is applied for measuring the nonlinear refractive index of ferrofluids. PMID- 21629314 TI - Half-data-page insertion method for increasing recording density in angular multiplexing holographic memory. AB - We have developed a method to use a half-size data page between two full-size data pages to increase the recording density in angular multiplexing holographic memory up to 1.5* as much as the conventional angular multiplexing sequence. In our recording sequence, the full- and half-size data pages are alternately multiplexed. This is because each plane wave from various points in a data page has different angular selectivity. A half-size data page has higher angular selectivity than a full-size data page. The required angular intervals were estimated by numerical simulation taking holographic medium tilt into account. Also, an angular multiplexing experiment using the half-data-page insertion method resulted in a low bit error rate of the order of 10(-3), which is sufficient for practical use. PMID- 21629315 TI - Tunable photonic bandgap in a one-dimensional superconducting-dielectric superlattice. AB - The transmittance of one-dimensional photonic crystals consisting of superconductor and lossless dielectric has been systematically studied through the transfer-matrix method. Obviously, the shift of the photonic bandgap (PBG) becomes more noticeable by adjusting the thicknesses of the dielectric layers than that of superconductor layers. Furthermore, the number of PBGs can be controlled by varying the thicknesses of dielectric layers. Compared to the thicknesses of the dielectric layers, the width of the PBGs is more sensitive to the thicknesses of the superconductor layers. However, the width of the first PBG promptly varies when the thicknesses of the dielectric layers increase from 0 to 40 nm. If the contribution of the normal conducting electrons of the superconductor is nonnegligible, the temperature of the superconductor has no influence on the width of the PBGs. Moreover, the damp coefficient does not affect the PBGs under low-temperature conditions. PMID- 21629316 TI - Fast-updating and nonrepeating Lissajous image reconstruction method for capturing increased dynamic information. AB - We present a fast-updating Lissajous image reconstruction methodology that uses an increased image frame rate beyond the pattern repeat rate generally used in conventional Lissajous image reconstruction methods. The fast display rate provides increased dynamic information and reduced motion blur, as compared to conventional Lissajous reconstruction, at the cost of single-frame pixel density. Importantly, this method does not discard any information from the conventional Lissajous image reconstruction, and frames from the complete Lissajous pattern can be displayed simultaneously. We present the theoretical background for this image reconstruction methodology along with images and video taken using the algorithm in a custom-built miniaturized multiphoton microscopy system. PMID- 21629317 TI - Maskless fabrication of three-dimensional microstructures with high isotropic resolution: practical and theoretical considerations. AB - A maskless three-dimensional (3D) microfabrication method based on a digital micromirror device (DMD) is proposed for high lateral and vertical resolution. A substrate is scanned laterally under virtual masks of the DMD. The masks are allocated to a large number of virtual slices, all of which are projected in a single scan of the stage. A theoretical model for the cumulative dose distribution in a photoresist is derived and used to predict the resulting 3D profile. Experiments showed that the proposed method is promising for avoiding the stair-step problem and preventing misalignment errors. PMID- 21629318 TI - Driving plasmonic nanoantennas with triangular lasers and slot waveguides. AB - Plasmonic nanoantennas can generate high-intensity electric fields in a very small area. However, being passive devices, they need to be excited by external laser sources. The excitation of nanoantennas by semiconductor lasers can be inefficient and a significant amount of light may return back to the laser source after being scattered by the nanoantenna. In this paper, it is shown that the amount of light being returned to the semiconductor laser can be reduced by using dielectric slot waveguides. These waveguides can transport the incident light to the nanoantennas, but the amount of nondirectional back-scattered light is reduced after propagation through the slot waveguide. PMID- 21629319 TI - Practical method to derive nonlinear response functions of cameras for scientific imaging. AB - We developed a practical method to derive response functions that convert the amount of incident light to the counts of analog-to-digital conversion (A/D) of cameras for scientific imaging. In this method, we need a mechanism to accurately control the amount of incident light into cameras just within a limited dynamic range and at a limited number of steps. A variable brightness light source, which supplies the incident light into cameras, is also necessary, but we do not need to know its accurate brightness. Thus, this method enables us to derive the nonlinear response functions accurately with such a simple setup. PMID- 21629320 TI - Measuring the dispersive properties of liquids using a microinterferometer. AB - Using a single-beam, compact interferometer, we measure the refractive index of liquids in the near IR. This highly compact device relies on a silica capillary with a 50 MUm inner diameter: it uses a minimal volume of test liquid, isolates the liquid from the humid atmosphere, has broadband operation, and is inherently mechanically stable. These characteristics, in combination with straightforward data acquisition, make it particularly well-suited for measuring the optical properties in the near IR of a wide range of liquids. Using this refractometer, we measure the refractive index of high-index liquids that are expected to be hydroscopic. The accuracy of the refractometer (+/-0.1%) is demonstrated through measuring the indices of air and pure water. We show that the hydroscopic behavior of the probed liquids has little influence on their optical properties in the near IR. PMID- 21629321 TI - Exact revision of the elliptically bent mirror theory. AB - One of the main hurdles for nanometer focusing by a bending mirror lies in the theoretical surface errors by its approximations used for the traditional theory. The impacts of approximations and analytical corrections have been discussed, and the elliptically bent mirror theory has been described during exact mathematical analysis without any approximations. These approximations are harmful for the focusing system with bigger grazing angle, bigger mirror length, and bigger numerical aperture. The properties of equal-moment and single-moment bent mirrors have been described and discussed. Because of its obvious advantages, a single moment bending mirror has high potential ability for nanometer focusing. PMID- 21629322 TI - Schwarzschild spectrometer. AB - This is a proposal and description of a new spectrometer based on the Schwarzschild optical system. The proposed design contains two Schwarzschild optical systems. Light diverging from the spectrometer entrance slit is collimated by the first one; the collimated light beam hits a planar diffraction grating and the light dispersed from the grating is focused by the second system, which is concentric with the first. A very simple procedure obtains designs that are anastigmatic for the center of the slit and for a particular wavelength. A specific example shows the performance of this type of spectrometer. PMID- 21629323 TI - Two-dimensional continuous wavelet transform for phase determination of complex interferograms. AB - A robust two-dimensional continuous wavelet transform (2D-CWT) technique for interferogram analysis is presented. To cope with the phase determination ambiguity issue encountered in the analysis of complex interferograms, a phase determination rule is proposed according to the phase distribution continuity, and a frequency-guided scheme is employed to obtain the correct phase distribution following a conventional 2D-CWT analysis. The theories are given in details, and the validity of the proposed technique is verified by computer simulation and real experiments. PMID- 21629324 TI - Bidirectional reflectance distribution function of Spectralon white reflectance standard illuminated by incoherent unpolarized and plane-polarized light. AB - A Lambert surface would appear equally bright from all observation directions regardless of the illumination direction. However, the reflection from a randomly scattering object generally has directional variation, which can be described in terms of the bidirectional reflectance distribution function (BRDF). We measured the BRDF of a Spectralon white reflectance standard for incoherent illumination at 405 and 680 nm with unpolarized and plane-polarized light from different directions of incidence. Our measurements show deviations of the BRDF for the Spectralon white reflectance standard from that of a Lambertian reflector that depend both on the angle of incidence and the polarization states of the incident light and detected light. The non-Lambertian reflection characteristics were found to increase more toward the direction of specular reflection as the angle of incidence gets larger. PMID- 21629325 TI - Comparison of two-, three-, and four-exposure quadrature phase-shifting holography. AB - In standard (four-exposure) quadrature phase-shifting holography (QPSH), two holograms and two intensity maps are acquired for zero-order-free and twin-image free reconstruction. The measurement of the intensity map of the object light can be omitted in three-exposure QPSH. Furthermore, the measurements of the two intensity maps can be omitted in two-exposure QPSH, and the acquisition time of the overall holographic recording process is reduced. In this paper we examine the quality of the reconstructed images in two-, three-, and four-exposure QPSH, in simulations as well as in optical experiments. Various intensity ratios of the object light and the reference light are taken into account. Simulations show that two- and three-exposure QPSH can provide reconstructed images with quality comparable to that of four-exposure QPSH at a low intensity ratio. In practice the intensity ratio is limited by visibility, and thus four-exposure QPSH exhibits the best quality of the reconstructed image. The uniformity and the phase error of the reference light are also discussed. We found in most cases there is no significant difference between the reconstructed images in two- and three-exposure QPSH, and the quality of the reconstructed images is acceptable for visual applications such as the acquisition of three-dimensional scene for display or particle tracking. PMID- 21629326 TI - Concentric ring metal grating for generating radially polarized light. AB - A subwavelength concentric ring metal grating for visible light (lambda=632.8 nm) is designed and fabricated by electron-beam lithography to transform circularly polarized light into radially polarized light. Experimental results are compared to theoretical predictions and the advantages and disadvantages of the element with alternative methods are discussed. PMID- 21629327 TI - Removal of single point diamond-turning marks by abrasive jet polishing. AB - Single point diamond turning (SPDT) is highly controllable and versatile in producing axially symmetric forms, non-axially-symmetric forms, microstructured surfaces, and free forms. However, the fine SPDT marks left in the surface limit its performance, and they are difficult to reduce or eliminate. It is unpractical for traditional methods to remove the fine marks without destroying their forms, especially for the aspheres and free forms. This paper introduces abrasive jet polishing (AJP) for the posttreatment of diamond-turned surfaces to remove the periodic microstructures. Samples of diamond-turned electroless nickel plated plano mirror were used in the experiments. One sample with an original surface roughness of more than 400 nm decreased to 4 nm after two iterations abrasive jet polishing; the surface roughness of another sample went from 3.7 nm to 1.4 nm after polishing. The periodic signatures on both of the samples were removed entirely after polishing. Contrastive experimental research was carried out on electroless nickel mirror with magnetorheological finishing, computer controlled optical surfacing, and AJP. The experimental results indicate that AJP is more appropriate in removing the periodic SPDT marks. Also, a figure maintaining experiment was carried out with the AJP process; the uniform polishing process shows that the AJP process can remove the periodic turning marks without destroying the original form. PMID- 21629328 TI - Relationship between the size distribution of mineral pigments and color saturation. AB - The saturation and the lightness of hematite layers are quantitatively related to the size distribution of the pigments, when applied on skin or a perfect white substrate. The optical properties, reflectance spectra, and colorimetric coordinates are calculated from the complex refractive index of hematite and by use of the radiative transfer equation. Monodisperse pigments are investigated first and the maximum of saturation is calculated as a function of the pigment radius. Polydisperse pigments are then investigated with a log-normal size distribution. The maximum of saturation is then calculated as a function of the width of the pigment distribution, for different mean radii. This modeling can be extended to any mineral pigments. PMID- 21629329 TI - Effective schema for the rigorous modeling of grating diffraction with focused beams. AB - Most modal diffraction methods are formulated for incident plane waves. In practical applications, the probing beam is focused. Usually, this is simulated by means of numerical integration where Gaussian quadrature formulas are most effective. These formulas require smooth integrands, which is not fulfilled for gratings due to Rayleigh singularities and physical resonances. The violation of this condition entails inaccurate integration results, such as kinks and other artifacts. In this paper, a methodology for the efficient treatment of the numerical integration with improved accuracy is presented. It is based on the subdivision of the aperture along the lines of Rayleigh singularities, mapping of these subapertures into unit squares, and separate application of the Gaussian cubature formulas for each subarea. PMID- 21629330 TI - Calculation and evaluation of narcissus for diffractive surfaces in infrared systems. AB - In infrared optical systems, the narcissus effect for diffractive surfaces should be calculated with specific diffraction orders based on the diffraction efficiency. It is shown in this work that the diffraction order of maximum diffraction efficiency varies with the change of the incident angle and wavelength of the backward-traced narcissus flux. Meanwhile, yni, which is the paraxial evaluation criterion of narcissus intensity for a refractive surface, is modified considering diffraction when a ray passes through diffractive surfaces, and a practical example has been given. The analysis can be used to calculate and control the narcissus intensity in infrared optical systems with diffractive surfaces. PMID- 21629331 TI - Sustainability and equity of renal replacement therapy in Spain. PMID- 21629332 TI - How to treat corticosteroid-resistant idiopathic focal segmental glomerulosclerosis? PMID- 21629333 TI - The role of mTOR inhibitors in renal diseases. PMID- 21629334 TI - Establishing and controlling chronic renal failure treatment costs. A pressing need. PMID- 21629335 TI - Effects of rapamycin on angiomyolipomas in patients with tuberous sclerosis. AB - BACKGROUND: Tuberous sclerosis (TS) is a systemic disease, with an autosomal dominant pattern of inheritance caused by mutations in two genes (TSC1 and TSC2) that cause tumours (angiomyolipomas [AML], angiofibromas, astrocytomas). Constant and inadequate proliferation occurring in TS may be blocked by mTOR inhibitors (mammalian target of rapamycin), such as rapamycin. MATERIAL AND METHODS: At present, our study includes 17 patients with TS. All had at least one AML greater than 2cm in diameter diagnosed by MRI. They received rapamycin during 12 months. Plasma levels remained stable between 4-8ng/dl. The AML size was monitored every six months by abdominal MRI. RESULTS: At 12 months of inclusion, MRI indicated a decrease in the size of AML in all patients showing at least a 50% reduction in 82.4% (14/17, 95% CI [56.57%, 96.20%]). The mean percent reduction was 66.3% (95% CI [56.9%, 75.6%], P<.0001). The major side effects observed were: oral aphthous ulcers (5/17); hypertriglyceridemia (3/17); microcytosis and hypochromia (3/17); diarrhea (2/17); acne (1/17); acute pyelonephritis (1/17); and proteinuria (1/17). CONCLUSIONS: These preliminary clinical data suggest that rapamycin can play a beneficial role in the treatment of TS. Our experience in 17 patients treated for 12 months demonstrates safety and efficacy in reducing AML volume. PMID- 21629336 TI - Multicentre study of haemodialysis costs. AB - BACKGROUND: Previous studies to determine the cost of haemodialysis (HD) in Spain have significant limitations: they are outdated or used indirect methods. There is also a lack of analysis performed simultaneously on Public centres (PC), with direct HD services, and partially state-subsidised centres (SC). This is an important issue since the two systems coexist in Spain. OBJECTIVES: To estimate the cost of HD replacement therapy for chronic renal failure in several centres. METHODS: This is a prospective and publicly-funded study, which estimates the costs for 2008 using a cost accounting system with specific allocation criteria. We collected demographic and comorbidity data for each centre. RESULTS: Six centres participated, two PC and four SC. There were no significant differences between centres in terms of patient demographics, time on haemodialysis and the Charlson comorbidity index. The total cost per patient per year ranged between ? 46, 254 and ? 33,130. The cost per patient per year (excluding vascular access and hospital admission) for PC was ? 42, 547 and ? 39, 289 and for SC ? 32 872, ? 29, 786, ? 35, 461 and ? 35, 294 (23% more in PC than SC). Costs related to staff/patient/year and consumables/patient/year were 67% and 83% respectively, higher for PC than SC. The highest percentage cost was for staff (average 30.9%), which showed significant variability between centres, both in absolute numbers (staff cost per patient per year between ? 18,151 and ? 8504) and as a percentage (between 42.6 % and 25.4%). CONCLUSIONS: Cost variability exists among different HD centres, and this can be attributed primarily to staff and consumables costs, which is higher for PC than SC. PMID- 21629337 TI - Factors associated with blood pressure control in diabetic patients treated in nephrology units. PRESDIAB Study. AB - BACKGROUND AND OBJECTIVE: Most hypertensive patients do not reach target blood pressure (BP), especially if they are diabetic. The objective of the study is to assess the percentage of tight BP control, defined as BP<130/80mm Hg and identify factors associated with it in diabetic type 2 (DM2) patients treated in nephrology units. PATIENTS AND METHODS: Observational and cross-sectional study; we included 526 patients with DM2 and arterial hypertension (AHT). We collected data on: demographics, anthropometrics, harmful habits, history of cardiovascular disease (CVD), blood pressure, kidney function, glycaemic control, lipid profile, and drug treatment, among others. RESULTS: The mean age (SD) was 66 (10.6) years, 61% were male, 12.8% were smokers, 39.4% had a history of CVD, 72% had hypercholesterolemia, and 44% were obese. Seventeen point five percent of patients had tight BP control (<130/80mm Hg) (95% confidence interval [CI]:14.3 21.0), while 36.9% had BP below 140/85mm Hg. Seventy-one percent of patients were prescribed two or more anti-hypertensive treatments. Several factors are associated with tight BP control not being achieved, and the logistic regression analysis revealed that LDL cholesterol levels were significantly associated (odds ratio [OR] 0.55; 95% CI:0.41-0.75 for one standard deviation increase). CONCLUSIONS: Of the DM2 patients that attended the nephrology units, less than 20% achieved a tight BP control. Cholesterol levels seem to be the main factor associated with unsatisfactory BP control within our study population. PMID- 21629338 TI - Impact of an interdisciplinary training course on counselling and decision making support for nephrology department professionals. AB - A 12-hour training program was delivered to the professionals of a nephrology department. Contents of the course were about difficult communication skills in health care interactions. Counselling was the relational methodology instructed. The objective was to assess changes in attitudes in relation with bioethics principles and knowledge. Variables were measured before and after the training program. Sample was composed by 76 professionals (57% nurses, 26% auxiliary nurses y 17% nephrologists) for knowledge and 27 professionals for variable attitudes. Considering the total sample, results show changes in implication with bioethics principles (p <0.05) and knowledge (p <0.001). There are differences related to the kind of profession. Nurses benefit more from the training program attending in the variable knowledge (p <0.001). PMID- 21629339 TI - Fuller Albright and our current understanding of calcium and phosphorus regulation and primary hyperparathyroidism. AB - The major contributions of Fuller Albright to our understanding of calcium and phosphorus regulation and primary hyperparathyroidism are highlighted. Albright was the first investigator to initiate a systematic study of mineral metabolism. With resources limited to the measurement of serum calcium and phosphorus and the infusion of parathyroid extract, Albright used balance studies to establish a framework for our understanding of calcium and phosphorus regulation and primary hyperparathyroidism. Albright was the first to show that the etiology of primary hyperparathyroidism could be from either an adenoma or hyperplasia of the parathyroid glands and stone disease was a separate manifestation of primary hyperparathyroidism. Albright also showed that: 1) a renal threshold for calcium excretion was present in hypoparathyroid patients; 2) correction of hypocalcemia in hypoparathyroid patients with vitamin D had a phosphaturic action; 3) renal failure reduced the intestinal absorption of calcium in primary hyperparathyroidism; 4) the ''hungry bone'' syndrome developed after parathyroidectomy in severe primary hyperparathyroidism; and 5) a target organ can fail to respond to a hormone. He also suggested that a malignant tumor could be responsible for ectopic hormone production. Finally, our review integrates the observations of Albright with our current knowledge of calcium regulation and disorders. PMID- 21629340 TI - Prevalence of chronic kidney disease and arteriosclerosis in a non-selected population. World Kidney Day. PMID- 21629341 TI - Monitoring sirolimus levels: how does it affect the immunoassay used? PMID- 21629342 TI - Good practice guidelines on the use of erythropoiesis-stimulating agents in 2011. PMID- 21629343 TI - Listeria monocytogenes: an infrequent cause of peritonitis in peritoneal dialysis. PMID- 21629344 TI - Arterial hypertension induced by pyeloureteral stenosis in horseshoe kidney. PMID- 21629345 TI - Successful treatment with sodium thiosulfate for calcific uraemic arteriolopathy. PMID- 21629346 TI - Spontaneous remission of nephrotic syndrome in a patient with diabetic nephropathy and Parkinson's disease. PMID- 21629347 TI - Immunotactoid glomerulopathy and tuberculosis: a novel association. PMID- 21629348 TI - Sarcoidosis: diagnosis from the renal function and hypercalcaemia study. PMID- 21629349 TI - Membranous glomerulonephritis in a patient with syphilis. PMID- 21629350 TI - Treatment with intravenous daptomycin for a peritonitis relapse caused by Staphylococcus epidermidis. PMID- 21629351 TI - Intraperitoneal daptomycin. PMID- 21629352 TI - Relapses in patients with microscopic polyangiitis with persistently positive antimyeloperoxidase for 4 years using maintenance immunosuppressants. PMID- 21629353 TI - Association between narcolepsy and H1N1 exposure. PMID- 21629355 TI - The demise of portable monitoring to diagnose OSA? Not so fast! PMID- 21629356 TI - A nuisance or nemesis: the adverse effects of snoring. PMID- 21629357 TI - An integrated health-economic analysis of diagnostic and therapeutic strategies in the treatment of moderate-to-severe obstructive sleep apnea. AB - STUDY OBJECTIVES: Obstructive sleep apnea (OSA) is a common disorder associated with substantially increased cardiovascular risks, reduced quality of life, and increased risk of motor vehicle collisions due to daytime sleepiness. This study evaluates the cost-effectiveness of three commonly used diagnostic strategies (full-night polysomnography, split-night polysomnography, unattended portable home-monitoring) in conjunction with continuous positive airway pressure (CPAP) therapy in patients with moderate-to-severe OSA. DESIGN: A Markov model was created to compare costs and effectiveness of different diagnostic and therapeutic strategies over a 10-year interval and the expected lifetime of the patient. The primary measure of cost-effectiveness was incremental cost per quality-adjusted life year (QALY) gained. PATIENTS OR PARTICIPANTS: Baseline computations were performed for a hypothetical average cohort of 50-year-old males with a 50% pretest probability of having moderate-to-severe OSA (apnea hypopnea index [AHI] >= 15 events per hour). MEASUREMENTS AND RESULTS: For a patient with moderate-to-severe OSA, CPAP therapy has an incremental cost effectiveness ratio (ICER) of $15,915 per QALY gained for the lifetime horizon. Over the lifetime horizon in a population with 50% prevalence of OSA, full-night polysomnography in conjunction with CPAP therapy is the most economically efficient strategy at any willingness-to-pay greater than $17,131 per-QALY gained because it dominates all other strategies in comparative analysis. CONCLUSIONS: Full-night polysomnography (PSG) is cost-effective and is the preferred diagnostic strategy for adults suspected to have moderate-to-severe OSA when all diagnostic options are available. Split-night PSG and unattended home monitoring can be considered cost-effective alternatives when full-night PSG is not available. PMID- 21629358 TI - Associations of Framingham risk score profile and coronary artery calcification with sleep characteristics in middle-aged men and women: Pittsburgh SleepSCORE study. AB - BACKGROUND: Short and less efficient sleep may be risk factors for atherosclerosis. Few studies have investigated the associations between sleep characteristics and early cardiovascular disease (CVD) risk. OBJECTIVE: Evaluate the associations between coronary artery calcification (CAC) and Framingham risk score profile with sleep characteristics in middle-aged men and women with no history of diagnosed myocardial infarction, interventional cardiology procedures, stroke, diabetes, or sleep disorders. METHOD: 224 participants enrolled in an epidemiological study of disparities in CVD risk were recruited for a 9-night assessment of sleep, with 2 nights of polysomnography (PSG) and 9 nights of actigraphy and sleep diaries. Of the 224 participants, 110 had high/moderate Framingham risk scores and 114 had low scores; 195 had computed tomography measures of CAC. RESULTS: Individuals who had any CAC or higher Framingham risk scores had elevated apnea/hypopnea index (AHI) values, independent of age, race, and gender. The AHI association with CAC was nonsignificant in analyses adjusting for body mass index (BMI). Those with higher Framingham risk score profiles had shorter PSG sleep duration and less percent stage 3-4 and delta power sleep. High blood pressure and left ventricular hypertrophy were related to AHI and sleep duration, independent of BMI. Neither sleep duration nor efficiency was associated with CAC. CONCLUSIONS: CAC was not associated with AHI, independent of BMI in a community-based sample of middle-aged men and women. Framingham risk score profiles were related to poor sleep. Sleep duration may not be related to early plaque burden in relatively healthy individuals. PMID- 21629359 TI - Upper airway collapsibility during wakefulness in children with sleep disordered breathing, as determined by the negative expiratory pressure technique. AB - STUDY OBJECTIVES: Upper airway (UA) collapsibility is a major factor in the pathophysiology of sleep disordered breathing (SDB). We hypothesized that the negative expiratory pressure (NEP) technique could distinguish between normal children and children with SDB even during wakefulness. DESIGN: During wakefulness, NEP of -5 and -10 cm H(2)O was applied during expiration in seated and supine positions. UA muscle activity (EMG) was measured using intra-oral electrodes. SETTING: Sleep laboratory. PARTICIPANTS: Twenty children with snoring, 20 with obstructive sleep apnea syndrome (OSAS), and 20 controls. MEASUREMENTS AND RESULTS: The ratio of the area under the expiratory flow-volume curve during NEP compared to tidal breathing (RatioNEP) was calculated. Similarly, EMG area under the curve during NEP as a ratio of baseline was measured (RatioEMG). There were significant differences in RatioNEP between controls and snorers and controls and OSAS, at both pressures, in both the seated and supine positions; P < 0.0001 for all (e.g., RatioNEP at -5 cm H(2)O, seated: 1.8 +/- 0.5, 2.1 +/- 0.4, and 3.0 +/- 0.6 for OSAS, snorers, and controls, respectively). However, no significant differences were found between snorers and OSAS. For RatioEMG, no significant differences were found between groups. CONCLUSIONS: RatioNEP distinguishes between normal children and children with SDB, be it snoring or OSAS, indicating that these children have a more collapsible UA even during wakefulness. However, it does not differentiate between snorers and OSAS, highlighting the important role of UA muscle activity during sleep. NEP technique does not elicit a different UA muscle activity response between controls and children with SDB. PMID- 21629360 TI - Baroreflex sensitivity during sleep in infants: impact of sleeping position and sleep state. AB - STUDY OBJECTIVES: The prone sleeping position is a major risk for the sudden infant death syndrome (SIDS) and has been associated with lowered blood pressure and impaired blood pressure control. This study aimed to assess the effects of sleeping position, sleep state, and postnatal age on baroreflex control of heart rate. PARTICIPANTS: Term infants (n = 31) were studied at 2-4 weeks, 2-3 months, and 5-6 months with daytime polysomnography. INTERVENTIONS: Blood pressure and heart rate were recorded during quiet (QS) and active (AS) sleep in both the supine and prone positions. In each condition, three 1-2 minute baseline measurements and three 15 degrees head-up tilts were performed. MEASUREMENTS AND RESULTS: Baroreflex sensitivity (BRS) was assessed using cross-spectral analysis (BRS(SP)) and sequence analysis (BRS(SEQ)) in the baseline condition and with BRS(SP) during head-up tilting (BRS(SP) Tilt). BRS was usually lower prone compared to supine, reaching significance at 2-3 months (BRS(SP), P < 0.05; BRS(SP) Tilt, P < 0.05) and 5-6 months (BRS(SEQ), P < 0.05). BRS was lower in AS than QS supine at 5-6 months for all BRS estimates (P < 0.05). During QS, BRS increased with postnatal age in both sleeping positions (P < 0.05 for all BRS estimates); during AS, the postnatal age-related increase was limited to the prone position (BRS(SEQ), P < 0.05). CONCLUSIONS: Sleeping position, sleep state and postnatal age all affect infant baroreflex function. Reduced BRS in the younger infants sleeping prone could increase the vulnerability to hypotensive events during sleep and thus play a vital role in conditions where circulatory failure may be involved, such as SIDS. PMID- 21629361 TI - Fewer spontaneous arousals in infants with apparent life-threatening event. AB - STUDY OBJECTIVES: A deficit in arousal process has been implicated as a mechanism of sudden infant death syndrome (SIDS). Compared with control infants, SIDS victims showed significantly more subcortical activations and fewer cortical arousals than matched control infants. Apparent life-threatening event (ALTE) is often considered as an aborted SIDS event. The aim of this study was to study the arousal characteristics of ALTE infants during the first months of life. DESIGN: 35 ALTE infants were studied with nighttime polysomnography at 2-3, 5-6, and 8-9 months of age. Eighteen of the infants had mothers who smoked. The infants were born full term and were usually supine sleepers. Sleep state and cardiorespiratory parameters were scored according to recommended criteria. Arousals were differentiated into subcortical activations or cortical arousals, according to the presence of autonomic and/or electroencephalographic changes. The results were compared with those of 19 healthy infants with nonsmoking mothers. RESULTS: During NREM sleep, the ALTE infants had fewer total arousals, cortical arousals, and subcortical activations at 2-3 and 5-6 months (P < 0.001) than control infants. ALTE infants with smoking mothers had more obstructive apnea (P = 0.009) and more subcortical activations during REM sleep at 2-3 months of age (P < 0.001) than ALTE infants with nonsmoking mothers. CONCLUSIONS: Spontaneous arousals were differently altered in ALTE infants than in SIDS infants, suggesting an entity different from SIDS. ALTE infants with smoking mothers had arousal and respiratory characteristics that were similar to future SIDS victims, suggesting some common abnormalities in brainstem dysfunction. PMID- 21629362 TI - The heterogeneity of obstructive sleep apnea (predominant obstructive vs pure obstructive apnea). AB - STUDY OBJECTIVES: To compare the breathing instability and upper airway collapsibility between patients with pure OSA (i.e. 100% of apneas are obstructive) and patients with predominant OSA (i.e., coexisting obstructive and central apneas). DESIGN: A cross-sectional study with data scored by a fellow being blinded to the subjects' classification. The results were compared between the 2 groups with unpaired student t-test. SETTING AND INTERVENTIONS: Standard polysomnography technique was used to document sleep-wake state. Ventilator in pressure support mode was used to introduce hypocapnic apnea during CO(2) reserve measurement. CPAP with both positive and negative pressures was used to produce obstructive apnea during upper airway collapsibility measurement. PARTICIPANTS: 21 patients with OSA: 12 with coexisting central/mixed apneas and hypopneas (28% +/- 6% of total), and 9 had pure OSA. MEASUREMENTS: The upper airway collapsibility was measured by assessing the critical closing pressure (Pcrit). Breathing stability was assessed by measuring CO(2) reserve (i.e., DeltaPCO(2) [eupnea-apnea threshold]) during NREM sleep. RESULTS: There was no difference in Pcrit between the 2 groups (pure OSA vs. predominant OSA: 2.0 +/- 0.4 vs. 2.7 +/- 0.4 cm H(2)O, P = 0.27); but the CO(2) reserve was significantly smaller in predominant OSA group (1.6 +/- 0.7 mm Hg) than the pure OSA group (3.8 +/- 0.6 mm Hg) (P = 0.02). CONCLUSIONS: The present data indicate that breathing stability rather than upper airway collapsibility distinguishes OSA patients with a combination of obstructive and central events from those with pure OSA. PMID- 21629363 TI - Tissue vibration induces carotid artery endothelial dysfunction: a mechanism linking snoring and carotid atherosclerosis? AB - STUDY OBJECTIVES: We have previously identified heavy snoring as an independent risk factor for carotid atherosclerosis. In order to explore the hypothesis that snoring-associated vibration of the carotid artery induces endothelial dysfunction (an established atherogenic precursor), we utilized an animal model to examine direct effects of peri-carotid tissue vibration on carotid artery endothelial function and structure. DESIGN: In supine anesthetized, ventilated rabbits, the right carotid artery (RCA) was directly exposed to vibrations for 6 h (peak frequency 60 Hz, energy matched to that of induced snoring in rabbits). Similarly instrumented unvibrated rabbits served as controls. Features of OSA such as hypoxemia, large intra-pleural swings and blood pressure volatility were prevented. Carotid endothelial function was then examined: (1) biochemically by measurement of tissue cyclic guanosine monophosphate (cGMP) to acetylcholine (ACh) and sodium nitroprusside (SNP); and (2) functionally by monitoring vessel relaxation with acetylcholine in a myobath. MEASUREMENT AND RESULTS: Vessel cGMP after stimulation with ACh was reduced in vibrated RCA compared with unvibrated (control) arteries in a vibration energy dose-dependent manner. Vibrated RCA also showed decreased vasorelaxation to ACh compared with control arteries. Notably, after addition of SNP (nitric oxide donor), cGMP levels did not differ between vibrated and control arteries, thereby isolating vibration-induced dysfunction to the endothelium alone. This dysfunction occurred in the presence of a morphologically intact endothelium without increased apoptosis. CONCLUSIONS: Carotid arteries subjected to 6 h of continuous peri-carotid tissue vibration displayed endothelial dysfunction, suggesting a direct plausible mechanism linking heavy snoring to the development of carotid atherosclerosis. PMID- 21629364 TI - Effects of stressor predictability and controllability on sleep, temperature, and fear behavior in mice. AB - STUDY OBJECTIVES: Predictability and controllability are important factors in the persisting effects of stress. We trained mice with signaled, escapable shock (SES) and with signaled, inescapable shock (SIS) to determine whether shock predictability can be a significant factor in the effects of stress on sleep. DESIGN: Male BALB/cJ mice were implanted with transmitters for recording EEG, activity, and temperature via telemetry. After recovery from surgery, baseline sleep recordings were obtained for 2 days. The mice were then randomly assigned to SES (n = 9) and yoked SIS (n = 9) conditions. The mice were presented cues (90 dB, 2 kHz tones) that started 5.0 sec prior to and co-terminated with footshocks (0.5 mA; 5.0 sec maximum duration). SES mice always received shock but could terminate it by moving to the non-occupied chamber in a shuttlebox. SIS mice received identical tones and shocks, but could not alter shock duration. Twenty cue-shock pairings (1.0-min interstimulus intervals) were presented on 2 days (ST1 and ST2). Seven days after ST2, SES and SIS mice, in their home cages, were presented with cues identical to those presented during ST1 and ST2. SETTING: NA. PATIENTS OR PARTICIPANTS: NA. INTERVENTIONS: NA. MEASUREMENTS AND RESULTS: On each training and test day, EEG, activity and temperature were recorded for 20 hours. Freezing was scored in response to the cue alone. Compared to SIS mice, SES mice showed significantly increased REM after ST1 and ST2. Compared to SES mice, SIS mice showed significantly increased NREM after ST1 and ST2. Both groups showed reduced REM in response to cue presentation alone. Both groups showed similar stress-induced increases in temperature and freezing in response to the cue alone. CONCLUSIONS: These findings indicate that predictability (modeled by signaled shock) can play a significant role in the effects of stress on sleep. PMID- 21629365 TI - Upper airway genioglossal activity in children with sickle cell disease. AB - STUDY OBJECTIVES: The prevalence of obstructive sleep apnea syndrome (OSAS) in sickle cell disease (SCD) has been reported to be higher than that in the general pediatric population. However, not all subjects with SCD develop OSAS. We hypothesized that SCD patients with OSAS have a blunted neuromuscular response to subatmospheric pressure loads during sleep, making them more likely to develop upper airway collapse. DESIGN: Subjects with SCD with and without OSAS underwent pressure-flow measurements during sleep using intraoral surface electrodes to measure genioglossal EMG (EMGgg). Two techniques were applied to decrease the nasal pressure (P(N)) to subatmospheric levels, resulting in an activated and relatively hypotonic upper airway. The area under the curve of the inspiratory EMGgg moving time average was analyzed. EMGgg activity was expressed as a percentage of baseline. Changes in EMGgg in response to decrements in nasal pressure were expressed as the slope of the EMGgg vs. nasal pressure (slope of EMGgg-P(N)). SETTING: Sleep laboratory. PARTICIPANTS: 4 children with SCD and OSAS and 18 children with SCD but without OSAS. RESULTS: THE MAJOR FINDINGS OF THIS STUDY WERE: (1) using the activated but not the hypotonic technique, the slope of EMGgg-P(N) was more negative in SCD controls than SCD OSAS; (2) the slope of EMGgg-P(N) was significantly lower using the activated technique compared to the hypotonic technique in SCD controls only; (3) similarly, the critical closing pressure, Pcrit, was more negative using the activated technique than the hypotonic technique in SCD controls but not in SCD OSAS. CONCLUSION: This preliminary study has shown that children with SCD but without OSAS have more prominent upper airway reflexes than children with SCD and OSAS. PMID- 21629366 TI - Sleep disturbances and nocturnal agitation behaviors in older adults with dementia. AB - STUDY OBJECTIVES: To examine nighttime sleep patterns of persons with dementia showing nocturnal agitation behaviors and to determine whether restless legs syndrome (RLS), periodic limb movements in sleep (PLMS), and obstructive sleep apnea (OSA) are associated with nocturnal agitation behaviors. DESIGN: Cross sectional. SETTING: General community. PARTICIPANTS: 59 participants with geriatrician-diagnosed dementia. Participants ages ranged from 66 to 88 years (mean age 79.1; SD 6.0). Mean Mini Mental State Examination (MMSE) score was 20.1 (SD 6.6). MMSE was used to measure baseline cognitive function and not for the diagnosis of dementia. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: Sleep was measured by 2 nights of in-home, attended, portable polysomnography (PSG). Nocturnal agitation was measured over 3 additional nights using the Cohen Mansfield Agitation Inventory modified for direct observations. Two experts independently and via consensus identified probable RLS. Total sleep time in participants was 5.6 h (SD 1.8 h). Mean periodic limb movements in sleep index (PLMI) was 15.29, and a high percentage (49%) had moderate to severe obstructive sleep apnea. Probable RLS was present in 24% of participants. Those with more severe cognitive impairment had longer sleep latency. Severe cognitive impairment, low apnea hypopnea index (AHI), and probable RLS were associated with nocturnal agitation behaviors (R(2) = 0.35, F(3,55) = 9.40, P < 0.001). CONCLUSIONS: It appears that probable RLS is associated with nocturnal agitation behaviors in persons with dementia, while OSA and PLMS are not. Further investigation is warranted to determine if treatment of RLS impacts nocturnal agitation behaviors in persons with dementia. PMID- 21629367 TI - Independent contributions of cortical gray matter, aging, sex and alcoholism to K complex amplitude evoked during sleep. AB - STUDY OBJECTIVES: The amplitude of the N550 component derived from the averaged evoked K-complex decreases with normal aging and with alcoholism. The study was designed to determine whether these declines are related to the extent of cortical or subcortical shrinkage. SETTING: Research sleep laboratory and MR imaging facility PARTICIPANTS: 26 abstinent long-term alcoholic men, 14 abstinent long-term alcoholic women, 18 control men, and 22 control women. MEASUREMENTS AND RESULTS: MRI data collected at 3T were analyzed from alcoholic and control men and women previously reported to have significantly different evoked delta activity during sleep. Segmented and parcellated MRI data collected at 3T were compared between these groups and evaluated for correlation with evoked K-complex amplitude measured at FP1, Fz, FCz, Cz, CPz, and Pz. Cortical gray matter and regional subcortical tissue volumes entered as predictors into stepwise multiple regression identified cortical gray matter as a unique significant predictor of evoked K-complex at all sites. Age added independent variance at 5 of the 6 sites, while alcoholism and sex added independent variance at frontal sites only. CONCLUSIONS: These data support recent intracranial studies showing cortical generation of K-complexes by indicating that cortical, but not subcortical volume contributes to K-complex amplitude. Establishing the extent of the relation between cortical volume and K-complex amplitude provides a mechanistic understanding of sleep compromise clinically relevant to normal aging, alcoholism, and likely other conditions affecting cortical volume and integrity. PMID- 21629368 TI - Time for bed: parent-set bedtimes associated with improved sleep and daytime functioning in adolescents. AB - STUDY OBJECTIVES: To determine the proportion of adolescents whose bedtime is set by their parents and to evaluate whether parent-set bedtimes are associated with earlier bedtimes, more sleep, and better daytime functioning. PARTICIPANTS: 385 adolescents aged 13-18 years (mean = 15.6, SD = 0.95; 60% male) from 8 socioeconomically diverse schools in South Australia. MEASUREMENTS AND METHODS: Adolescents completed the School Sleep Habits Survey during class time and then completed an 8-day Sleep Diary. The Flinders Fatigue Scale was completed on the final day of the study. RESULTS: 17.5% of adolescents reported a parent-set bedtime as the main factor determining their bedtime on school nights. Compared to adolescents without parent-set bedtimes, those with parent-set bedtimes had earlier bedtimes, obtained more sleep, and experienced improved daytime wakefulness and less fatigue. They did not differ significantly in terms of time taken to fall asleep. When parent-set bedtimes were removed on weekends, sleep patterns did not significantly differ between groups. CONCLUSIONS: Significant personal and public health issues, such as depression and accidental injury and mortality, are associated with insufficient sleep. Converging biological and psychosocial factors mean that adolescence is a period of heightened risk. Parent set bedtimes offer promise as a simple and easily translatable means for parents to improve the sleep and daytime functioning of their teens. PMID- 21629369 TI - Air leak is associated with poor adherence to autoPAP therapy. AB - STUDY OBJECTIVES: To our knowledge, a systematic study of the effect of air leak on adherence to auto-titrating positive airway pressure (autoPAP) therapy has not been reported. We hypothesized that in patients with obstructive sleep apnea (OSA), greater levels of air leak were associated with poor adherence to autoPAP therapy. DESIGN: Retrospective cohort study SETTING: Academic Center PARTICIPANTS: Ninety-six consecutive patients with high probability for OSA. INTERVENTIONS: N/A. MEASUREMENTS: Patients with OSA received 1 week of autoPAP therapy following which both adherence data and air leak information was downloaded from the device. Continuous positive airway pressure (CPAP) therapy was issued for a 5-week period with pressure determined by 90th percentile of that delivered during autoPAP therapy. Adequate adherence was defined as average usage > 4 h per night on all nights. RESULTS: Forty-three patients were adherent to autoPAP therapy (350 +/- 67[SD] min/day), whereas 53 patients were not (122 +/ 65 min/day; P < 0.0001). Air leak that was adjusted for pressure delivered was greater in non-adherent patients (7.0 +/- 3.5 L/min/cm H(2)O) than that in adherent patients (4.9 +/- 1.7 L/min/cm H(2)O; P < 0.0001). Greater residual respiratory events (measured as autoPAP-derived hypopnea index) and proportion of time spent at large leak levels were associated with non-adherence. Patients who were adherent to autoPAP therapy received higher average therapeutic pressures from the autoPAP device than non-adherent patients. Multivariate logistic regression revealed that higher levels of air leak were associated with non adherence to autoPAP therapy (odds ratio 1.43; 95% CI, 1.03, 1.98; P = 0.03). Moreover, adherence to autoPAP therapy was strongly correlated with subsequent adherence to CPAP therapy (R(2) = 0.74; P < 0.0001). CONCLUSION: Air leak was associated with poor adherence to autoPAP therapy. We speculate that air leak could be a potential target for future studies aimed at enhancing adherence to autoPAP therapy. PMID- 21629370 TI - Electroacupuncture for residual insomnia associated with major depressive disorder: a randomized controlled trial. AB - STUDY OBJECTIVES: To evaluate the efficacy and safety of electroacupuncture as an additional treatment for residual insomnia associated with major depressive disorder (MDD). DESIGN: Randomized, placebo-controlled. SETTING: A psychiatric outpatient clinic. PARTICIPANTS: 78 Chinese patients with DSM-IV-diagnosed MDD, insomnia complaint, a Hamilton Rating Scale for Depression (HDRS(17)) score <= 18, and fixed antidepressant dosage. INTERVENTION: Electroacupuncture, minimal acupuncture (superficial needling at non-acupuncture points), or noninvasive placebo acupuncture 3 sessions weekly for 3 weeks. MEASUREMENTS AND RESULTS: Insomnia Severity Index (ISI), Pittsburgh Sleep Quality Index (PSQI), HDRS(17), 1 week sleep diaries, and 3 day actigraphy were administered at baseline, 1 week post-treatment, and 4 week post-treatment. There was significant group by time interaction in ISI, PSQI, and sleep diary-derived sleep efficiency (mixed-effects models, P = 0.04, P = 0.03, and P = 0.01, respectively). Post hoc pairwise comparisons revealed that electroacupuncture and minimal acupuncture were more efficacious than placebo acupuncture in ISI and PSQI at 1 week and 4 week post treatment. Minimal acupuncture resulted in greater improvement in sleep diary derived sleep efficiency than placebo acupuncture at 1 week post-treatment. There was no significant between-group difference in actigraphy measures, depressive symptoms, daily functioning, and hypnotic consumption, and no difference in any measures between electroacupuncture and minimal acupuncture. CONCLUSION: Compared with placebo acupuncture, electroacupuncture and minimal acupuncture resulted in greater improvement in subjective sleep measures at 1 week and 4 week post treatment. No significant difference was found between electroacupuncture and minimal acupuncture, suggesting that the observed differences could be due to nonspecific effects of needling, regardless of whether it is done according to traditional Chinese medicine theory. PMID- 21629371 TI - The use of botulinum toxin A in children with cerebral palsy, with a focus on the lower limb. AB - PURPOSE: The purpose of this review is to clarify the role of botulinum toxin serotype A (BTX-A) in the treatment of children with cerebral palsy (CP), with a special focus on the lower limb. BACKGROUND: The treatment of spasticity is central in the clinical management of children with CP. BTX-A blocks the release of acetylcholine at the motor end plate, causing a temporary muscular denervation and, in an indirect way, a reduced spasticity. Children with increased tone develop secondary problems over time, such as muscle contractures and bony deformities, which impair their function and which need orthopaedic surgery. However in these younger children, delaying surgery is crucial because the results of early surgical interventions are less predictable and have a higher risk of failure and relapse. As BTX-A treatment reduces tone in a selective way, it allows a better motor control and muscle balance across joints, resulting in an improved range of motion and potential to strengthen antagonist muscles, when started at a young age. The effects are even more obvious when the correct BTX-A application is combined with other conservative therapies, such as physiotherapy, orthotic management and casts. There is now clear evidence that the consequences of persistent increased muscle tone can be limited by applying an integrated multi-level BTX-A treatment approach. Nevertheless, important challenges such as patient selection, defining appropriate individual goals, timing, dosing and dilution, accuracy of injection technique and how to measure outcomes will be questioned. Therefore, "reflection is more important than injection" remains an actual statement. PMID- 21629372 TI - Randomized controlled trial comparing above- and below-elbow plaster casts for distal forearm fractures in children. AB - PURPOSE: Long arm cast is the method of immobilization after closed reduction of the fracture of the distal third of the forearm, although short arm cast has been used to immobilize the forearm by some orthopedic surgeons. We conducted this study to evaluate the rate of displacement, union time, complication, and cost of treatment between the above-elbow and below-elbow plaster cast groups. METHODS: We designed a prospective randomized controlled trial to compare above- and below elbow casts for patients of distal forearm fracture aged 4-12 years. One hundred and one patients were presented to our institution during the period from 1st June 2007 to 31st May 2008. Twelve patients were excluded from the study. Eighty nine patients were randomized into two groups. Four patients were lost to follow up. Hence, 85 patients, 42 in the above-elbow group and 43 in the below-elbow group, were analyzed. Informed consent was obtained prior to participation in the trial. The parameters to be studied were defined before treatment and noted in the pilot-tested pro forma and follow up was done. RESULTS: Among the 85 patients analyzed, randomization was successful. Pain and swelling on the next day and at one week following cast application was significantly higher (P = 0.000) in the above-elbow group. Three cases in the above-elbow group needed slitting of the plaster cast. Remanipulation was needed more often in the above-elbow group (9.5%). Although the cast index was not statistically significant (P = 0.054) between the groups, those who needed remanipulation had a larger cast index (0.80). The total cost of the treatment and the number of school days lost was significantly higher (P = 0.000) in the above-elbow group. CONCLUSION: Below elbow cast treatment was comparable in terms of redisplacement, union time, and movement of the wrist to the above-elbow treatment. The complications rate and total cost was higher in the above-elbow plaster group, which could be a cost benefit if a below-elbow cast is used. PMID- 21629373 TI - No effect of osteopathic treatment on trunk morphology and spine flexibility in young women with adolescent idiopathic scoliosis. AB - INTRODUCTION: Brace treatment is the gold standard for patients with mild adolescent idiopathic scoliosis (Cobb angle 20 degrees -40 degrees ). However, negative psychosocial impacts, physical constraints and incompliance cause many patients and parents to seek for so-called holistic and apparently less harmful approaches within the field of complementary and alternative medicine (CAM). Osteopathy-manual interventions on the viscera and locomotor system-is widely used for scoliosis. There is, however, a complete lack of evidence regarding its efficacy. We, therefore, tested the hypothesis that osteopathy alters trunk morphology, a prerequisite to unload the concave side of the scoliosis, and that it halts curve progression. METHODS: This was a prospective, controlled trial of 20 post-pubertal young women (20 degrees -40 degrees idiopathic scoliosis) randomly allocated to an observation (group 0) or osteopathic treatment (group 1). The latter comprised three sessions (5 weeks). Trunk morphology (clinical examination, video rasterstereography) and spine flexibility (MediMouse((r))) were assessed at a pre- and post-intervention with a 3-month interval (blinded examiner). We chose scoliometer measurement (rib hump, lumbar prominence) as the main outcome parameter. RESULTS: Two patients in the treatment group refused further treatment and the final examination, as they felt no benefit after two osteopathic treatments. Regression analysis for repeat measurements (independent statistician) revealed no therapeutic effect on rib hump, lumbar prominence, plumb line, sagittal profile and global spinal flexibility. CONCLUSIONS: We found no evidence to support osteopathy in the treatment of mild adolescent idiopathic scoliosis. Therefore, we caution against abandoning the conventional standard of care for mild idiopathic scoliosis. As for other CAM therapies, the use of osteopathy as a treatment option for scoliosis still needs to be clearly defined. PMID- 21629374 TI - Objective compliance of adolescent girls with idiopathic scoliosis in a dynamic SpineCor brace. AB - PURPOSE: Dynamic SpineCor was designed to overcome the disadvantages of rigid orthoses-bulkiness, physical constraint and warming-and to improve the acceptance and compliance as limiting factors of brace treatment. Those theoretical benefits have not been proved by compliance studies yet. METHODS: SpineCor braces of 12 adolescent scoliotic girls were equipped with flexible temperature loggers, which were sandwiched between gel foam and the brace's pelvic base. Patients and parents were blinded for the observation period of 14 days and gave post-hoc consent. The logger yielded 672 time-stamped values at 30-min intervals. Pilot testing revealed values beyond the 30 degrees C threshold as indicative for brace wear. RESULTS: The average overall compliance (% wearing hours/prescribed 23 h) was 54% +/- 22.3 (range, 11.8-95.8%). The youngest patients (aged 10-12 years) were significantly more compliant than the others (P < 0.05). Most patients had a scattered wearing pattern: one was a day-wearer, one a night-wearer and only the two high adherers showed a consistent daily pattern. There was no significant difference between weekdays, weekdays and weekends, nor between day and night wear. CONCLUSION: The current study showed that the compliance of patients in a dynamic SpineCor is as limited as in a conventional brace. This is in line with earlier data on patients' SpineCor and rigid brace acceptance evaluated by a questionnaire and on temperature logging in rigid braces. PMID- 21629375 TI - Can children with spina bifida and muscular dystrophy participate in their own health status evaluations? AB - BACKGROUND: The outcomes movement has emphasized the importance of the patient in evaluating treatment outcome. However, concern has been raised about the ability of children, particularly those with multisystem disease, to evaluate their disability. PURPOSE: To determine whether children with spina bifida and muscular dystrophy and their parents agree when asked about the relative difficulty of daily activities and the severity of symptoms experienced by the child. METHODS: First, a list of symptoms and activity restrictions was generated from the literature, clinicians, and interviews with families with spina bifida and muscular dystrophy. Second, another group of parents and children with spina bifida (with hip dislocation or scoliosis) and with muscular dystrophy (with scoliosis), including those before and after surgery, independently rated the severity and importance of their objective and subjective complaints. RESULTS: The correlation between parents and their children was high for both objective (median Spearman's = 0.70; standard deviation [SD] = +/-0.17; range = -0.05-1.00) and subjective (median Spearman's = 0.76; SD = +/-0.14; range = 0.13-1.00) complaints, with an overall excellent level of agreement (Kappa = 0.75; 95% confidence interval [CI]: 0.73, 0.76). CONCLUSION: Children with spina bifida and muscular dystrophy are capable of understanding and assessing their disability. PMID- 21629376 TI - Results of bipolar release in the treatment of congenital muscular torticolis in patients older than 10 years of age. AB - PURPOSE: Neglected congenital muscular torticollis (CMT) is a common presentation in developing countries like India, with the primary concern of the patients being cosmesis. Little has been published regarding the management of such patients. The aim of our study was to evaluate the results of bipolar release in this age group. METHODS: Over a period of five years, 14 patients older than ten years (range 10-19 years) with neglected CMT were operated on with bipolar release of sternocleidomastoid muscle and z-plasty lengthening of the sternal end. Postoperative protocol included head halter traction for three weeks followed by intensive physical therapy. Results were evaluated using a modified version of the system of Lee et al. RESULTS: At an average follow-up of around three years (range 1-5 years), excellent results were noted in three patients, good in seven, fair in two, and poor in two. Postoperative improvements in range of motion, head tilt, chin deviation and cosmesis were noted in all patients, and these improvements were statistically significant. No surgery-related complications or recurrences requiring surgery occurred in any of the patients. CONCLUSIONS: This study concluded that patients with CMT presenting after ten years of age definitely benefit from surgery, and that bipolar release is an adequate and complication-free method for such patients. PMID- 21629377 TI - Treatment of femoral fractures in children: is titanium elastic nailing an improvement over hip spica casting? AB - PURPOSE: The purpose of the study was to assess the validity of surgical interference with elastic nailing in treating pediatric femur fractures in comparison with the traditional treatment method-hip spica casting. METHODS: Sixteen consecutive femur fractures in children 5-15 years of age were recruited prospectively over 13 months. An equal number of age-matched children treated by spica casting were recruited retrospectively. Subtrochanteric, supracondylar femur fractures were excluded. RESULTS: Fracture union occurred earlier in the surgical group (6 weeks) than in the spica group (8 weeks) (P = 0.001). Spica casting caused higher coronal plane angulation (P = 0.001), higher rotational malalignment (P < 0.001), higher limb length discrepancy at 1-year follow-up (P < 0.001), longer duration of immobilization (P < 0.001), later full weight-bearing (P < 0.001), and greater absence from school (P < 0.001). Flynn outcome scores were better with titanium elastic nailing than with hip spica casting. CONCLUSION: Titanium elastic nailing led to better outcomes compared to hip spica casting in terms of earlier union, lower rates of malunion, shorter rehabilitation milestones, and better functional outcome scores. PMID- 21629378 TI - Ilizarov hip reconstruction without external fixation: a new technique. AB - PURPOSE: The Ilizarov hip reconstruction is a well accepted but complication prone operative salvage procedure in chronically dislocated hips, not least due to the long-term application of external fixation. Although the advantages of fully implantable devices are well known in limb lengthening and are described consistently, until now, external fixation has been used exclusively to perform the Ilizarov hip reconstruction procedure. We present a new technique of Ilizarov hip reconstruction with purely internal implants. METHODS: A 14-year-old girl with a history of spina bifida presented with a 4-cm-short right leg, a Trendelenburg gait and a complex neurological disease expression. Because of refusal of external fixation by the patient and significantly lower complication rates, an Ilizarov hip reconstruction without external fixation was performed. A locking compression plate was applied to fix the proximal femoral valgus extension osteotomy and a motorised intramedullary distraction nail was used for the distal, lengthening-varisation osteotomy. RESULTS: A healing index of 33 days/cm and full weight bearing after 6 months were noted. At the 1 year follow up, the patient showed an improvement of the Trendelenburg gait, as well as successful leg equalisation. Satisfaction to a high degree was additionally noted by factors such as reduced pain, the ability to wear workaday clothes and cosmetically appealing scars. No complications were recorded. CONCLUSION: The exclusive use of internal implants for Ilizarov hip reconstruction is a feasible and patient-friendly alternative to traditional methods. Their use, however, may be restricted by geometric preconditions. PMID- 21629379 TI - Lateral mass screw fixation in children. AB - PURPOSE: The safety and feasibility of posterior screw fixation of the cervical spine in children has not been well documented in the orthopedic literature. We performed a retrospective review of our experience using posterior cervical screw fixation in children. METHODS: The medical records and radiologic records of 36 children at a mean age of 10 years (range 3-16 years) were reviewed. Diagnoses included: ten instability, 11 deformity, seven trauma, five tumor, and three congenital abnormalities. Operative reports and postoperative computed tomography (CT) scans were reviewed to determine the technical feasibility of screw placement, any screw-related complications, and to assess for correct screw position. In this series, there were no neurologic complications, no vertebral artery injuries, and no screw-related complications. RESULTS: Thirty patients (141 screws) had screws evaluated postoperatively and were shown to be completely contained on postoperative CT scans. There were no revisions due to screw failure or dislodgement. There were no vascular or neurologic complications. CONCLUSIONS: Posterior screw fixation in the pediatric population may be done safely and greatly enhances fixation strength for a variety of disorders requiring instrumentation and fusion. PMID- 21629380 TI - Corpectomy and circumferential spinal fusion in dystrophic neurofibromatous curves. AB - PURPOSE: Spinal deformity in neurofibromatosis, when associated with dystrophic change, is a major treatment challenge. Early anterior and posterior spinal fusion is the accepted treatment plan for severe progressive kyphoscoliosis. Resection of the vertebral body as an additional step in correction and fusion has not been previously investigated. Information on the functional status of the neurofibromatous curves is also scarce in the literature. The purpose of this study is to report an experience with corpectomy and circumferential fusion in dystrophic spines of neurofibromatosis. METHODS: Among 16 patients with dystrophic spinal curves, nine cases who had undergone anterior and posterior fusion with 6.7 years (range 2-11.9) average follow-up were evaluated clinically, radiographically, and functionally (SRS-22). RESULTS: Nine cases with a mean age of 11.8 years (range 7.8-17) at surgery consisted of seven kyphoscoliotic patients, who had received one or two levels of cord decompression and corpectomy, and two cases of scoliosis with hypokyphosis. Surgery improved the mean preoperative scoliosis of 87 (range 60-110) and local kyphosis of 69.3 degrees (range 50-100) to 49 (range 15-85) and 49 degrees (range 35-70), respectively. Loss of correction of 5 degrees in the scoliosis and 13 degrees in the kyphosis angles was observed in the final follow-up. One surgery resulted in an immediate postoperative mortality. Fusion was achieved in all cases. The average SRS-22 score of 4.1 (range 3-4.6) was obtained. CONCLUSIONS: Simultaneous anterior and posterior fusion with corpectomy for dystrophic neurofibromatous spinal deformities is associated with a high fusion rate, good correction, and also good functional outcome. PMID- 21629381 TI - Cruelty in maternity wards: fifty years later. AB - Fifty years have passed since a scandal broke over inhumane treatment of laboring women in U.S. hospitals, yet first-person and eyewitness reports document that medical care providers continue to subject childbearing women to verbal and physical abuse and even to what would constitute sexual assault in any other context. Women frequently are denied their right to make informed decisions about care and may be punished for attempting to assert their right to refusal. Mistreatment is not uncommon and persists because of factors inherent to hospital social culture. Concerted action on the part of all stakeholders will be required to bring about systemic reform. PMID- 21629382 TI - Lamaze certified childbirth educators: drawing strength from our past to build the future. AB - This year's 50th anniversary of Lamaze International is a time to reflect upon our past and present as we work together to build the future of childbirth. In the 1950s and 1960s, thoughtful men and women such as Elisabeth Bing began to look carefully at the birthing practices in the United States. Lamaze Certified Childbirth Educators became leaders in improving birthing practices by teaching women and their partners the truth about how women were cared for during childbirth. Currently, the rise in maternal mortality and morbidity in the United States illustrates the pressing need for more changes. Lamaze Certified Childbirth Educators are leading change by pushing for the wide adoption of Lamaze International's Six Healthy Birth Practices to promote natural, safe, and healthy birth. PMID- 21629383 TI - Birth in an ordinary instant. AB - Our daily lives are a series of ordinary moments and unnoticed thresholds-times that define us in ways we often do not give much attention. While we consider childbirth to be one of life's extra-ordinary events, the hours of labor and birth need not be dramatic (or traumatic) ones. I describe a quiet, well supported birth in the Netherlands that is cause for celebration of the beauty of an ordinary instant that can define and enrich the human experience. PMID- 21629384 TI - Women's Desire for Pregnancy. AB - Descriptive qualitative interviews were conducted with 16 women who had given birth in the previous 5 years to identify factors that influenced their desire to become pregnant. All interviews were audiotaped. Following the interviews, the texts were transcribed verbatim and analyzed using Giorgi's method. Five themes emerged from the interviews: (1) timing; (2) spacing; (3) meeting personal criteria; (4) desire for the experience of pregnancy, birth, and parenting; and (5) extended family in close proximity. The study's findings offer an initial step in understanding the attitudes, motivations, and beliefs of healthy childbearing women and their desire for pregnancy. Increased understanding may help health-care providers develop interventions that not only assist women to conceive as desired but also prevent unplanned pregnancies. PMID- 21629385 TI - Down Memory Lane: Recollections of Lamaze International's First 50 Years. AB - The 42-year involvement of one member of Lamaze International is chronicled through a decade-by-decade review of personal memories. The history of Lamaze International is shared through the recollections of her roles as a childbirth educator, faculty member, and member of the board of directors. PMID- 21629386 TI - Revisiting the classics in childbirth education. AB - In celebration of Lamaze International's 50th anniversary, reviewers share their thoughts on some classic Lamaze resource materials and how their content relates to today's viewpoints on birth and childbirth education. Although some of the material may be outdated, all of the resources offer timeless insights as well as a unique view on the history of childbirth education. The following topics are addressed: past cultural views of birth; advocacy for change in birth practices; Lamaze method; pioneers in childbirth; importance of childbirth education; and birth advocacy. PMID- 21629387 TI - How do women who plan home birth prepare for childbirth? AB - In this column, the findings of a secondary analysis of data from a larger qualitative study of the experience of home birth are discussed. The aim was to describe the ways in which women who plan home birth prepare for their births. The findings provide support for the idea of birth preparation and education occurring throughout pregnancy and describe the ways in which women planning to give birth at home develop confidence, plan for support, and make decisions related to the particulars of the labor and birth. Implications of these findings for childbirth education are explored. PMID- 21629388 TI - A call to come together: why it matters and why it works. AB - This letter to the editor is in response to Barbara Hotelling's column, "Tools for Teaching - The Power of Numbers: Transforming Birth Through Collaborations," published in the Volume 19, Number 2 (Spring 2010) issue of The Journal of Perinatal Education. Hotelling's brief overview of the history of birth activism in the United States and of current collaborations among diverse groups and individuals provides inspiration to continue efforts that improve maternity care and promote natural, safe, and healthy childbirth practices. PMID- 21629389 TI - Steps toward innovative childbirth education: selected strategies from the blueprint for action. AB - To mark the 50th anniversary of Lamaze International, Childbirth Connection celebrates landmark accomplishments in education for childbearing women and families, and takes stock of the changing educational needs and preferences of current childbearing families in looking toward the future. Childbirth Connection's multi-year, multi-stakeholder Transforming Maternity Care initiative resulted in two landmark reports: 2020 Vision for a High-Quality, High-Value Maternity Care System and Blueprint for Action: Steps Toward a High-Quality, High Value Maternity Care System. Selected recommendations of greatest relevance to the field of childbirth education are discussed, and the new Transforming Maternity Care Partnership is introduced. PMID- 21629390 TI - What once was old is new again. AB - Through seminars and continuing educational opportunities, recently certified Lamaze childbirth educators beginning their teaching careers have learned of the changes in birth practices over the past 50 years. However, they may not have heard the personal stories about the locations and conditions in which Lamaze educators first taught. In this column, five childbirth educators share their memories of the birthing climate, teaching strategies, class populations, and other aspects of Lamaze childbirth education 50 years ago and how the "old ways" compare with today's classes. Their stories not only provide an important, historical perspective to build on the past and improve future educational opportunities for expectant women and their families but also illustrate Lamaze's ongoing efforts to promote natural, safe, and healthy birth practices. PMID- 21629391 TI - Patients' perspectives on the role of prepared childbirth education in decision making regarding elective labor induction. AB - The purpose of our study was to explore reasons why nulliparous women chose to have an elective labor induction and to identify the influence of prepared childbirth classes on their decision. The study included 1,349 nulliparous women at term who participated in a survey regarding their choices for childbirth, their attendance at prepared childbirth classes, and their experience with labor and birth. Sixty-three percent of women who attended childbirth classes and did not have elective induction reported that classes provided helpful information to assist in their decision-making process. Study results suggest attendance at prepared childbirth classes can be an effective source of information regarding elective labor induction and influential in women's decisions regarding whether or not to have elective labor induction. Women perceive prepared childbirth classes positively and find the information provided valuable. PMID- 21629392 TI - Lose and gain: impacts of ERK5 and JNK cascades on each other. AB - Kinase cascades in ERK5 (Extracellular signal-regulated kinases) and JNK (c-Jun N terminal kinases) signaling pathways mediate the sensing and processing of stimuli. Cross-talks between signaling cascades is a likely phenomenon that can cause apparently different biological responses from a single pathway, on its activation. Feedback loops have the potential to greatly alter the properties of a pathway and its response to stimuli. Based on enzyme kinetic reactions, mathematical models have been developed to predict and analyze the impacts of cross-talks and feedback loops in ERK5 and JNK cascades. It has been observed that, there is no significant impact on neither ERK5 activation nor JNKs' activation due to cross-talks between them. But it is due to cross-talks and feedback loops in ERK5 and JNK cascade, ERK5 gets activated in a transient manner in the absence of input signals. Planning to obtain the parameter values from the experimentalist and the result should be validated by experimental verification. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11693-010-9061-4) contains supplementary material, which is available to authorized users. PMID- 21629393 TI - Abdominal and back pain in a 65-year-old patient with metastatic prostate cancer. AB - OBJECTIVE: Prostate cancer remains the second leading cause of cancer-related deaths, and African American men are affected with this disease disproportionately in terms of incidence and mortality. The purpose of this article is to present a case report that illustrates the importance of a careful evaluation, including a comprehensive historical review and appropriate physical and laboratory assessment, of a patient with back pain and seemingly unrelated symptoms. CLINICAL FEATURES: A 65-year-old African American man presented to a chiropractic clinic after experiencing lower back pain for 1 month. The digital rectal examination was unremarkable, but the serum prostate-specific antigen was markedly elevated. A suspicion of metastatic prostate cancer resulted in subsequent referral, further diagnostic evaluation, and palliation. INTERVENTION AND OUTCOME: The patient was referred for medical evaluation and palliation of his condition. Spinal decompression surgery of the thoracic spine was initiated, resulting in weakness and paresthesia in the lower limbs bilaterally. The patient died because of the complications associated with the medical interventions and the disease about 12 months after the referral. CONCLUSION: Chiropractic physicians should maintain a high degree of suspicion for catastrophic causes of back-related complaints, such as metastatic prostate cancer. The Prostate Cancer Prevention Trial Risk Calculator, a research validated instrument, should be used in the assessment of prostate cancer risk. Performance of the digital rectal examination and of the prostate-specific antigen determination remains integral in the clinical assessment of the health status in aging men, with or without back pain. PMID- 21629394 TI - Resolution of low back and radicular pain in a 40-year-old male United States Navy Petty Officer after collaborative medical and chiropractic care. AB - OBJECTIVE: The aim of this study is to describe the interdisciplinary care, including chiropractic services, in a military health care facility of an active duty member of the United States Navy with low back pain, leg pain, and foot numbness. CLINICAL FEATURES: A 40-year-old patient developed low back pain, leg pain, and foot numbness after moving furniture. The patient described his symptoms as pain in the right low back, pain shooting into the right lateral thigh and lower leg, and numbness into the right lateral foot. Magnetic resonance imaging confirmed disk extrusion at L4/L5 occupying the lateral recess and abutting the exiting right L5 nerve root. INTERVENTION AND OUTCOME: Providers, including primary care, chiropractic, and orthopedics, in an established multidisciplinary health care system contributed to the case management. The patient received 11 chiropractic treatments (spinal manipulation, flexion distraction, abdominal rehab exercises) over 72 days. Subjective complaints resolved, and the patient was released back to full duty. CONCLUSION: Integrative care, using medical and chiropractic services, was successful in the conservative management of a patient with low back pain and radicular symptoms secondary to disk extrusion. PMID- 21629395 TI - Chiropractic management of a 40-year-old female patient with Meniere disease. AB - OBJECTIVE: The purpose of this case report is to describe the chiropractic management of a patient with Meniere disease. CLINICAL FEATURES: A 40-year-old woman presented with a diagnosis of Meniere disease including a 2-month history of vertigo and a 16-month history of left-sided tinnitus, low-frequency hearing loss, and aural fullness. The patient's other symptoms included left-sided neck pain, temporomandibular joint pain, and headaches. Examination revealed left sided upper cervical joint dysfunction along with myofascial trigger points in the middle and upper trapezius muscle. INTERVENTION AND OUTCOME: Treatment included primarily high-velocity, low-amplitude spinal manipulation to the upper cervical and thoracic spine, along with soft-tissue trigger-point therapy, and stretching exercises. Within 2 weeks of treatment, the patient's tinnitus had resolved; and all other symptoms (including vertigo) were improved. The patient's headaches, neck pain, and vertigo were subsequently resolved within 3 months of treatment. The patient experienced only 2 minor episodes of self-resolving "light headedness" over that time. After 21/2 years of follow-up, any occasional episodes of mild aural fullness and/or light-headedness are either self-resolving or relieved with cervical spinal manipulation and soft-tissue treatment. CONCLUSION: This case report suggests that chiropractic care, including upper cervical spinal manipulation and soft-tissue therapy, may be beneficial in treating some patients with Meniere disease. PMID- 21629396 TI - Chiropractic management of a 5-year-old boy with urinary and bowel incontinence. AB - OBJECTIVE: The purpose of this article is to describe chiropractic management of a 5-year-old boy with urinary and bowel incontinence. CLINICAL FEATURES: A 5-year old boy presented with the primary symptoms of a complete lack of bowel and bladder control with prior surgical correction for lumbar meningocele, spinal lipoma, and tethered spinal cord. Examination revealed spinal and pelvic dysfunction. INTERVENTION AND OUTCOME: Chiropractic treatment methods included using the Activator adjusting instrument and shortwave diathermy to the lumbar spine and sacrum. A total of 5 treatments were initially provided over a period of 4 weeks. After the initial treatment period, he was able to maintain satisfactory control of his bladder and bowel, day and night, for a period of approximately 6 months. A second course of treatments was initiated approximately 6 months later because of a recurrence of bladder and bowel incontinence. Four additional treatments were provided over a period of 4 weeks. This second course of treatment reestablished satisfactory control of bladder and bowel function. CONCLUSION: For this patient, chiropractic care was successful in establishing satisfactory bladder and bowel control. PMID- 21629397 TI - Community-acquired methicillin-resistant Staphylococcus aureus: a potential diagnosis for a 16-year-old athlete with knee pain. AB - OBJECTIVE: This case report aims to raise awareness in chiropractic physicians of the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in adolescents who participate in sports activities and to alert the chiropractic physician of the necessity to consider potential diagnoses that are not within their typical clinical heuristic. CLINICAL FEATURES: A 16-year-old adolescent girl entered the clinic with a complaint of left knee pain that had an insidious onset during her involvement in sports activities. Later that same day, her knee became enlarged, red, and had pustular formations with a discharge. She was taken to an urgent care facility and subsequently diagnosed with MRSA. Her history included treatment of a left knee musculoskeletal condition 6 weeks prior to which she had responded favorably. INTERVENTIONS AND OUTCOMES: She was treated medically with an aggressive course of antibiotic therapy and excision of the furuncle. The chiropractic physician played a role in patient education and notifying local school authorities of the case. CONCLUSION: Doctors of chiropractic must prepare themselves for the unexpected and remain open to diagnostic possibilities outside of the normal scope of practice. Knee pain or cellulitis of any type may require additional diagnostic and patient care protocols to make the correct diagnosis. With the incidence of community-acquired MRSA increasing at an alarming rate, it is certainly a diagnosis doctors of chiropractic should be aware of when treating patients, especially those involved in sports activities. PMID- 21629398 TI - Clinical detection of abdominal aortic aneurysm in a 74-year-old man in chiropractic practice. AB - OBJECTIVE: The purpose of this article is to present a case of abdominal aortic aneurysm to illustrate its clinical detection through history and physical examination and the importance of this condition to the chiropractic clinical setting. CLINICAL FEATURES: A 74-year-old retired man consulted a doctor of chiropractic for chronic low back pain. The history and physical examination confirmed chronic sacroiliac and a lumbar facet dysfunction. After 5 weeks, the patient stated he had stomach cramps. After this, a more thorough abdominal examination was done. The doctor of chiropractic detected an enlarged pulsatile mass upon abdominal palpation. INTERVENTION AND OUTCOME: The patient was sent to the cardiologist and had successful surgery within weeks. CONCLUSION: An abdominal aortic aneurysm has specific symptoms and associated risk factors. If known risk factors are present, a clinical examination needs to be carried out, even though sensitivity of the clinical examination may be low. It should be a differential diagnosis in every male patient older than 50 years with low back pain. In case of suspicion, the patient should be referred for advanced imaging. PMID- 21629399 TI - Heterointegration of Pt/Si/Ag Nanowire Photodiodes and Their Photocatalytic Properties. AB - Photocatalyst mediated photoelectrochemical processes can make use of the photogenerated electrons and holes onsite for photocatalytic redox reactions, and enable the harness and conversion of solar energy into chemical energy, in analogy to natural photosynthesis. However, the photocatalysts available to date are limited by either poor efficiency in the visible light range or insufficient photoelectrochemical stability. We show that a Pt/Si/Ag nanowire heterostructure can be rationally synthesized to integrate a nanoscale metal-semiconductor Schottky diode encased in a protective insulating shell with two exposed metal catalysts. The synthesis of Pt/Si/Ag nanowire diodes involves a scalable process including the formation of silicon nanowire array through wet chemical etching, electrodeposition of platinum and photoreduction of silver. We further demonstrated that the Pt/Si/Ag diodes exhibited highly efficient photocatalytic activity for a wide range of applications including environmental remediation and solar fuel production in the visible range. In this article, photodegradation of indigo carmine and 4-nitrophenol were used to evaluate the photoactivity of Pt/Si/Ag diodes. The Pt/Si/Ag diodes also show high activity for photoconversion of formic acid into carbon dioxide and hydrogen. PMID- 21629400 TI - Informed consent. PMID- 21629401 TI - Embolization of musculoskeletal bone tumors. AB - Bone tumors may present as incidental findings, with pain or loss of function, or as fractures. There is a broad range of indications for transarterial embolization (TAE) in primary or metastatic bone tumors: to reduce operative hemorrhagic risks, to simplify or allow more definitive surgery, or in the context of pain palliation, fever, bleeding, or hypercalcemic and other rheological factors. Embolization may also increase tumor sensitivity to chemotherapy or radiation therapy. The procedure itself is often complex with significant risk to adjacent structures and is usually part of a wider treatment strategy. There are many options of embolic agent, techniques, and end points but all aim to devascularize the tumor. Catheter angiography at the time of TAE is used to determine the correct embolic agent and technique with care taken to isolate at risk structures. Many factors determine the best choice of embolic material, probably the most important of which is operator experience. In life threatening situations or in preoperative embolizations of metastatic tumors, many operators opt for a combination of particulate emboli and stainless steel or platinum coils. Agents discussed include polyvinyl alcohol particles, trisacryl microspheres, gelatin sponge, liquid embolic agents, and embolization coils. Tumor types treated include vascular metastatic lesions, commonly renal cell or thyroid, particularly in locations prone to fracture; giant cell tumors; aneurysmal bone cysts; vertebral hemangiomas, osteosarcomas; arteriovenous malformations; and osteoblastomas. TAE should be considered in the treatment algorithm of primary or secondary bone tumors. Specific benefit is present where there is a high risk of bleeding at surgery, where there is spinal involvement and neural encroachment, where active bleeding is present or in awkward surgical locations where prolonged surgery is anticipated. PMID- 21629402 TI - Percutaneous bone tumor management. AB - Interventional radiology plays a major role in the management of bone tumors. Many different percutaneous techniques are available. Some aim to treat pain and consolidate a pathological bone (cementoplasty); others aim to ablate tumor or reduce its volume (sclerotherapy, thermal ablation). In this article, image guided techniques of primary and secondary bone tumors with vertebroplasty, ethanol injection, radiofrequency ablation, laser photocoagulation, cryoablation, and radiofrequency ionization (coblation) will be reviewed. For each modality, the principles, the indications, and the results will be presented. The technical choice depends on the therapeutic intent-curative or palliative-and the need for consolidation, but also on the general status of the patient and the other therapeutic options. For the most complex cases, combined treatments can be required. However, the less disabling technique should always be considered first. PMID- 21629403 TI - Percutaneous cementoplasty. AB - Cementoplasty includes percutaneous procedures like vertebroplasty, kyphoplasty, osteoplasty, and sacroplasty. Bone packing with cement aims to treat or prevent vertebral and extraspinal pathological fractures and relieve pain in patients with osteoporosis and bone metastases. The authors outline the accepted and newer indications for patient selection and present the fundamentals of image-guided lesion access and cement injection. Practitioners should evaluate each patient carefully and have a thorough knowledge of the anatomy, the technique, the expected outcomes, and the potential complications. Detailed informed consent and multidisciplinary decision making are recommended. Understanding of the particular advantages and limitations of the various modern filler materials is also crucial for a successful and uncomplicated procedure. Future developments include new mechanical devices for effective restoration of vertebral height, as well as the introduction of osteoconductive and osteoinductive cements that will be able to promote more physiological bone healing. PMID- 21629404 TI - Percutaneous treatment of intervertebral disc herniation. AB - Interventional radiology plays a major role in the management of symptomatic intervertebral disc herniations. In the absence of significant pain relief with conservative treatment including oral pain killers and anti-inflammatory drugs, selective image-guided periradicular infiltrations are generally indicated. The precise control of needle positioning allows optimal distribution of steroids along the painful nerve root. After 6 weeks of failure of conservative treatment including periradicular infiltration, treatment aiming to decompress or remove the herniation is considered. Conventional open surgery offers suboptimal results and is associated with significant morbidity. To achieve minimally invasive discal decompression, different percutaneous techniques have been developed. Their principle is to remove a small volume of nucleus, which results in an important reduction of intradiscal pressure and subsequently reduction of pressure inside the disc herniation. However, only contained disc herniations determined by computed tomography or magnetic resonance are indicated for these techniques. Thermal techniques such as radiofrequency or laser nucleotomy seem to be more effective than purely mechanical nucleotomy; indeed, they achieve discal decompression but also thermal destruction of intradiscal nociceptors, which may play a major role in the physiopathology of discal pain. The techniques of image guided spinal periradicular infiltration and percutaneous nucleotomy with laser and radiofrequency are presented with emphasis on their best indications. PMID- 21629405 TI - Diagnostic and therapeutic joint injections. AB - Joint injection is a useful tool in the diagnosis of intra-articular pathology that may improve diagnostic performance of computed tomography (CT) and magnetic resonance (MR) imaging. Historically, conventional arthrography under fluoroscopy was the first method to be used to image indirectly the intra-articular soft tissues, but with the advent of CT, CT arthrography offered better soft tissue depiction. The development of conventional MR allowed even better visualization of soft tissues, and in the early 1990s, MR arthrography surpassed CT arthrography in popularity. Joint injections may also be performed for therapeutic reasons with different drugs, such as corticosteroids, anesthetics, or hyaluronic acid, which have been shown to provide pain relief in various circumstances. In this article, the technical principles for joint injection of the shoulder, knee, elbow, hip, ankle, and wrist, used for therapeutic or diagnostic reasons, are discussed. Indications, expected benefits, and risks are also analyzed. PMID- 21629406 TI - Kyphoplasty. AB - The advent of the use of kyphoplasty in 1998 was the result of the expected evolution of materials and methods related to the successful introduction of vertebroplasty as a valid treatment of painful vertebral compression fractures. Kyphoplasty introduced a method of creating bilateral bone voids and in many cases elevation (reduction) of depressed end plates with variable degrees of height restoration. This was achieved by using bilateral balloon bone tamps capable of pressures significantly higher than conventional angioplasty balloons. This allows creation of bilateral bone voids, resulting in the ability to apply a much thicker cement mixture. This is felt to be the reason for fewer cement related complications compared with vertebroplasty including extravasation and embolization. Although the procedural cost presently is higher for kyphoplasty, this is expected to decrease as patents expire and industry competition increases. Kyphoplasty indications include all those of vertebroplasty plus additional ones either contraindicated or not recommended for treatment with vertebroplasty. Kyphoplasty achieves the same degree of pain relief as vertebroplasty but may offer additional benefits of fewer complications, more indications, better biopsy specimens, and potential for height restoration of compression fractures of the spine. Further studies regarding potential benefits are warranted to assess any added value of kyphoplasty compared with vertebroplasty. PMID- 21629407 TI - Diagnosis and planning in the management of musculoskeletal tumors: surgical perspective. AB - The evaluation of musculoskeletal tumors requires a close interaction between the orthopedic oncologist, radiologist, and the pathologist. Successful outcome can be achieved in a considerable number of patients by following the appropriate diagnostic strategies and staging studies. The aim of this article is to outline the presentation, imaging, and staging of the primary and metastatic bone and soft tissue tumors. Some of the image-guided interventions for these tumors are also presented. PMID- 21629408 TI - Image-guided musculoskeletal biopsies. AB - Image-guided musculoskeletal (MSK) biopsies are safe and effective procedures that yield diagnostic accuracies up to 97%. When performed in conjunction with a multidisciplinary team, they provide crucial information that will affect patient care and outcome. Computed tomography and ultrasound are the main modalities used to carry out MSK biopsies, and various needles and techniques are available to help the radiologist perform these procedures safely. PMID- 21629409 TI - Osteoplasty: Percutaneous Bone Cement Injection beyond the Spine. AB - Percutaneous osteoplasty, the injection of bone cement into a painful bone lesion refractory to conventional therapy (radiotherapy, chemotherapy, and narcotic analgesia), is performed to allow for immediate bone structure consolidation, reduce the risk of a pathological fracture, achieve pain regression, and improve mobility. In this article, the technique of this procedure and a review of the current literature including the author's personal large series will be described. PMID- 21629411 TI - Fluoroscopically guided balloon dilation of the esophagus. PMID- 21629412 TI - Hyperkalemia, Bradycardia, and Cardiac Arrest during Percutaneous Declotting of an Arteriovenous Graft. PMID- 21629410 TI - A Step-by-Step Practical Approach to Imaging Diagnosis and Interventional Radiologic Therapy in Vascular Malformations. AB - Within vascular anomalies, vascular malformations are those present at birth that grow with the patient and exhibit abnormal dilated vascular channels lined by mature endothelium. Vascular tumors, the other group of vascular anomalies, demonstrate endothelial hypercellularity. Vascular malformations are further divided into low-flow varieties (capillary, venous, and lymphatic malformations) and high-flow varieties (arteriovenous malformation and fistula). All malformations exhibit a predictable group of clinical patterns that vary in severity and rate of progression. The interventional radiologist must incorporate this clinical data with characteristic ultrasound and magnetic resonance findings to arrive at a diagnosis. One must then decide in a multidisciplinary fashion, based on objective clinical criteria and image-based morphology, if the patent is a candidate for intervention. Sclerotherapy is a technique used to treat vascular malformations whereby an endothelial-cidal agent is introduced into the endoluminal compartment to initiate vascular closure. The high flow rate of an arteriovenous malformation requires the incorporation of superselective transarterial, direct, and transvenous access with flow reduction techniques to deliver adequate dose of sclerosant and embolic to the nidus. Satisfactory outcomes are seen in over half of all malformations patients. Similar treatment related complications are seen between malformations but are lowest in lymphatic and highest in arteriovenous malformations. PMID- 21629413 TI - Castration of cats on-farm - A comment. PMID- 21629416 TI - Year of the Cat! Vet2011! 125th Anniversary of FMV! PMID- 21629417 TI - An ethicist's commentary on promoting farm animal welfare at the expense of productivity. PMID- 21629418 TI - The effects of exposure of susceptible alpacas to alpacas persistently infected with bovine viral diarrhea virus. AB - Reports of bovine viral diarrhea virus (BVDV) infections in alpacas have been increasing in recent years but much is still unknown about the mechanisms of disease in this species. This report characterizes the transmission of BVDV from persistently infected (PI) alpacas to BVDV naive alpacas, documents shedding patterns, and characterizes the disease effects in both PI and transiently infected alpacas. Two PI alpacas shed BVDV Type 1b virus in most body fluids, and commonly available diagnostic tests verified their status. Bovine viral diarrhea virus Type 1b transient infections produced only mild signs of disease in BVDV naive alpacas. Viremia was detected in whole blood, but viral shedding during the acute phase was not detected and antibody appeared to be protective upon re exposure to the virus. PMID- 21629419 TI - Decreased mortality of weaned pigs with Streptococcus suis with the use of in water potassium penicillin G. AB - This study evaluated the efficacy of potassium penicillin G in drinking water of weaned pigs to reduce mortality and spread of infection caused by Streptococcus suis. A total of 896 18-day-old weaned pigs were randomly assigned to either treatment with potassium penicillin G in-water (Treated), or no treatment (Control). The outcomes analyzed were total mortality, mortality due to S. suis, and overall counts of S. suis colonies. The risk of mortality due to S. suis and total mortality were significantly increased in the Control group compared with Treated pigs (P < 0.05). Bacterial culture of posterior pharyngeal swabs indicated that Control pigs were significantly more likely to have >= 1000 colonies of S. suis per plate than were Treated pigs (P < 0.05). This study demonstrates that potassium penicillin G administered in drinking water is effective in reducing mortality associated with S. suis infection and reducing tonsillar carriage of S. suis. PMID- 21629420 TI - Treatment of feline otitis externa due to Otodectes cynotis and complicated by secondary bacterial and fungal infections with Oridermyl auricular ointment. AB - A blinded randomized study was conducted on 24 cats to confirm the presence of bacterial and/or fungal secondary infections associated with otoacariasis and to verify the efficacy of Oridermyl, an acaricidal/antibiotic/antifungal/anti inflammatory ointment, for treatment of the primary infestation and secondary infections. Sixteen cats were treated once daily for 10 d; 4 cats were not treated and 4 were treated with a placebo ointment. On Days 0 and 10, ears were swabbed for counts of bacteria and yeasts, for bacterial culture and sensitivity, and examined for determination of the degree of clinical otitis. Auricular secretions were removed for mite counts on Day 10, except for 8 treated cats that were done on Day 30. There was a high number of bacteria and yeasts in most cats and Oridermyl treatment significantly decreased those numbers. Staphylococci were the most frequently isolated bacteria. No live ear mites were found in cats treated with Oridermyl or the placebo ointment. PMID- 21629421 TI - Cataracts in 44 dogs (77 eyes): A comparison of outcomes for no treatment, topical medical management, or phacoemulsification with intraocular lens implantation. AB - Outcomes for 77 cataractous eyes were compared after each eye underwent no treatment, topical medical treatment only, or phacoemulsification with intraocular lens implantation. Median follow-up time for all dogs was 2.3 y. Failure occurred in all untreated eyes and the rate of failure was 65 and 255 times higher than in medically and surgically treated eyes, respectively. The failure rate was 4 times higher in dogs receiving only medical treatment compared with dogs that received surgery. Across all groups, the success rate for mature and hypermature cataracts was lower than for immature cataracts. Regardless of cataract stage, the chance of success was higher for eyes undergoing phacoemulsification than for eyes that received medical management only. Results of this study support prompt referral for phacoemulsification when cataracts are diagnosed in dogs or, if referral is not possible, topical anti-inflammatory therapy. PMID- 21629422 TI - Transcatheter glue arterial embolization of a mass in the hind limb of a dog. AB - This article reports the use of transarterial glue embolization in the treatment of a soft-tissue mass in the hind limb of a dog that was referred for a > 15-cm diameter soft tissue mass in the caudal thigh. Clinical improvement showed that the percutaneous therapeutic cyanoacrylate glue embolization procedure was technically feasible and useful. PMID- 21629423 TI - Concurrent nephrosplenic entrapment and acquired inguinal herniation of the jejunum in a Standardbred stallion. AB - A stallion presenting for surgical correction of an acquired inguinal hernia was also diagnosed with a nephrosplenic entrapment (NSE) intraoperatively. Surgical intervention resulted in a successful outcome. To date, these conditions have not been reported to occur simultaneously. PMID- 21629424 TI - Autonomic dysfunction in a Jack Russell terrier. AB - A 4-year-old Jack Russell terrier was presented with an array of clinical signs suggestive of autonomic dysfunction. Many of the clinical signs were consistent with a diagnosis of dysautonomia; however, both chronicity and resolution of signs contradicted a diagnosis of this disease. PMID- 21629425 TI - Use of an osteoconductive compound as an aid in the management of a maxillary fracture in a boa constrictor. AB - A boa constrictor was presented with a short oblique compound fracture of the rostral third of the right maxilla. The fracture was reduced and biomaterial was placed around the fracture. A computed tomography scan at 1.5 mo post-surgery showed that the fracture had healed with slight displacement of the bone fragments. PMID- 21629426 TI - Potassium chloride as a euthanasia agent in psittacine birds: Clinical aspects and consequences for histopathologic assessment. AB - Twelve parrots anesthetized with isoflurane were euthanized intravenously (IV) with 3 or 10 mEq/kg body weight (BW) of potassium chloride (KCl) resulting in ventricular asystole at 68.0 s and 32.8 s, respectively. Mild vocalization (1/6 birds, 3 mEq/kg BW) and involuntary muscle tremors (5/6 birds, 10 mEq/kg BW) were noted. Unlike barbiturates or T-61 no histologic artefacts resulted from this technique. PMID- 21629427 TI - The use of nylon cable ties to repair rib fractures in neonatal foals. AB - Commercially available nylon cable ties were used as a method of internal fixation for displaced rib fractures in 8 equine neonates. The procedure was effective, well-tolerated, rapid, and inexpensive. The use of nylon cable ties as internal fixation for displaced rib fractures in foals is an effective surgical procedure. PMID- 21629428 TI - Imaging of exogenous lipoid pneumonia simulating lung malignancy in a dog. AB - This report presents peculiar radiographic and computed tomographic features of a lung mass in a dog, later diagnosed by histology as exogenous lipoid pneumonia. Although lipoid pneumonia is a well-known pathological entity in pets, this is the first imaging description of a single infiltrative lesion mimicking a neoplasm. PMID- 21629429 TI - Congenital nasolacrimal atresia in 4 alpacas. AB - Four alpacas, 2 wk to 1 y of age, were diagnosed with congenital atresia of the nasal puncta of the nasolacrimal duct. Dacryocystorhinography confirmed and localized the atresia. All animals were treated successfully by surgical creation of nasal puncta and patency was ensured by placement of a surgical stent for several weeks. PMID- 21629430 TI - Gallbladder mucocele in a 12-year-old cocker spaniel. AB - A 12-year-old, spayed cocker spaniel was presented for panting behavior which had increased over the past several months. A diagnosis of hyperadrenocorticism was made, and ultrasound imaging revealed the presence of a gallbladder mucocele. While often an incidental finding, gallbladder mucoceles can have consequences and require intervention. The etiology, diagnosis, and management of gallbladder mucoceles are discussed. PMID- 21629431 TI - From print to screen: Female veterinarians in memoir and film. PMID- 21629432 TI - Diagnostic ophthalmology. PMID- 21629433 TI - Development of calorie restriction mimetics as therapeutics for obesity, diabetes, inflammatory and neurodegenerative diseases. AB - Calorie restriction (CR) is the most robust intervention that decreases morbidity and mortality, and thereby increases the lifespan of many organisms. Although the signaling pathways involved in the beneficial effects of CR are not yet fully understood. Several candidate pathways and key molecules have been identified. The effects of CR are highly conserved from lower organisms such as yeast to higher mammals such as rodents and monkeys. Recent studies have also demonstrated beneficial effects of CR in humans, although we need much longer studies to evaluate whether CR also increases the lifespan of humans. In reality, it is difficult for us to conduct CR interventions in humans because the subjects must be kept in a state of hunger and the duration of this state needed to achieve a clinically meaningful effect is still unknown. Thus, research in this field is focusing on the development of molecules that mimic the beneficial effects of CR without reducing food intake. Some of these candidate molecules include plant derived functional chemicals (phyto-chemicals), synthetic small molecules, and endocrine molecules such as adipokines. Several studies have already shown that this research field may yield novel drugs for the treatment of age-related diseases such as diabetes. In this article, we describe the target pathways, candidate molecules, and strategies to develop CR mimetics. PMID- 21629434 TI - Cytosine methyltransferases as tumor markers. AB - Changes in DNA methylation patterns is a prominent characteristic of human tumors. Tumor cells display reduced levels of genomic DNA methylation and site specific CpG island hypermethylation. Methylation of CpG dinucleotides is catalyzed by the enzyme family of DNA methyltransferases (DNMTs). In this review, the role of DNA methylation and DNMTs as key determinants of carcinogenesis is further elucidated. The chromatin modifying proteins that are known to interact with DNMTs are also described. Finally, the role of DNMTs as potential therapeutic targets is addressed. PMID- 21629435 TI - Dihydrofolate reductase gene variations in susceptibility to disease and treatment outcomes. AB - Dihydrofolate reductase (DHFR) catalyzes the reduction of dihydrofolate to tetrahydrofolate (THF). THF is needed for the action of folate-dependent enzymes and is thus essential for DNA synthesis and methylation. The importance of this reaction is demonstrated by the effectiveness of antifolate medications used to treat cancer by inhibiting DHFR, thereby depleting THF and slowing DNA synthesis and cell proliferation. Due to the pivotal role that DHFR plays in folate metabolism and cancer treatment, changes in the level of DHFR expression can affect susceptibility to a variety of diseases dependent on folate status such as spina bifida and cancer. Likewise, variability in DHFR expression can affect sensitivity to anti-cancer drugs such as the folate antagonist methotrexate. Alterations in DHFR expression can be due to polymorphisms in the DHFR gene. Several variations have recently been described in DHFR, including promoter polymorphisms, the 19-bp deletion allele and variations in 3'UTR. These polymorphisms seem to be functional, affecting mRNA levels through various interesting mechanisms, including regulation through RNA interference. Several groups have assessed the association of these polymorphisms with folate levels, risk of cancer and spina bifida as well as the outcome of diseases treated with MTX. The latter may lead to different treatment schedules, improving treatment efficacy and/or allowing for a reduction in drug side effects. This review will summarize present knowledge regarding the predictive potential of DHFR polymorphisms in disease and treatment. PMID- 21629436 TI - Role of NFAT5 in inflammatory disorders associated with osmotic stress. AB - Nuclear factor of activated T cells 5 (NFAT5) is the most recently described member of the Rel family of transcription factors, including NF-kappaB and NFAT1 4, which play central roles in inducible gene expression during the immune response. NFAT5 was initially described to drive osmoprotective gene expression in renal medullary cells, which are routinely faced by high extracellular osmolalities. Recent data however indicate profound biological importance of the mammalian osmotic stress response in view of NFAT5 dependent gene regulation in non-renal tissues. In mononuclear cells and epithelial cells, NFAT5 stimulates the expression of various pro-inflammatory cytokines during elevated ambient tonicity. Accordingly, compared to plasma, the interstitial tonicity of lymphoid organs like spleen and thymus and that of liver is substantially hypertonic under physiological conditions. In addition, anisotonic disorders (hypernatremia, diabetes mellitus, dehydration) entail systemic hyperosmolality, and, in inflammatory disorders, the skin, intestine, and cornea are sites of local hyperosmolality. This article summarizes the current knowledge regarding systemic and local osmotic stress in anisotonic and inflammatory disorders in view of NFAT5 activation and regulation, and NFAT5 dependent cytokine production. PMID- 21629437 TI - Genetics of interleukin 1 receptor-like 1 in immune and inflammatory diseases. AB - Interleukin 1 receptor-like 1 (IL1RL1) is gaining in recognition due to its involvement in immune/inflammatory disorders. Well-designed animal studies have shown its critical role in experimental allergic inflammation and human in vitro studies have consistently demonstrated its up-regulation in several conditions such as asthma and rheumatoid arthritis. The ligand for IL1RL1 is IL33 which emerged as playing an important role in initiating eosinophilic inflammation and activating other immune cells resulting in an allergic phenotype.An IL1RL1 single nucleotide polymorphism (SNP) was among the most significant results of a genome wide scan investigating eosinophil counts; in the same study, this SNP associated with asthma in 10 populations.The IL1RL1 gene resides in a region of high linkage disequilibrium containing interleukin 1 receptor genes as well as interleukin 18 receptor and accessory genes. This poses a challenge to researchers interested in deciphering genetic association signals in the region as all of the genes represent interesting candidates for asthma and allergic disease.The IL1RL1 gene and its resulting soluble and receptor proteins have emerged as key regulators of the inflammatory process implicated in a large variety of human pathologies We review the function and expression of the IL1RL1 gene. We also describe the role of IL1RL1 in asthma, allergy, cardiovascular disease, infections, liver disease and kidney disease. PMID- 21629438 TI - Retinoic Acid Induced 1, RAI1: A Dosage Sensitive Gene Related to Neurobehavioral Alterations Including Autistic Behavior. AB - Genomic structural changes, such as gene Copy Number Variations (CNVs) are extremely abundant in the human genome. An enormous effort is currently ongoing to recognize and catalogue human CNVs and their associations with abnormal phenotypic outcomes. Recently, several reports related neuropsychiatric diseases (i.e. autism spectrum disorders, schizophrenia, mental retardation, behavioral problems, epilepsy) with specific CNV. Moreover, for some conditions, both the deletion and duplication of the same genomic segment are related to the phenotype. Syndromes associated with CNVs (microdeletion and microduplication) have long been known to display specific neurobehavioral traits. It is important to note that not every gene is susceptible to gene dosage changes and there are only a few dosage sensitive genes. Smith-Magenis (SMS) and Potocki-Lupski (PTLS) syndromes are associated with a reciprocal microdeletion and microduplication within chromosome 17p11.2. in humans. The dosage sensitive gene responsible for most phenotypes in SMS has been identified: the Retinoic Acid Induced 1 (RAI1). Studies on mouse models and humans suggest that RAI1 is likely the dosage sensitive gene responsible for clinical features in PTLS. In addition, the human RAI1 gene has been implicated in several neurobehavioral traits as spinocerebellar ataxia (SCA2), schizophrenia and non syndromic autism. In this review we discuss the evidence of RAI1 as a dosage sensitive gene, its relationship with different neurobehavioral traits, gene structure and mutations, and what is known about its molecular and cellular function, as a first step in the elucidation of the mechanisms that relate dosage sensitive genes with abnormal neurobehavioral outcomes. PMID- 21629439 TI - Functional genomics of brain aging and Alzheimer's disease: focus on selective neuronal vulnerability. AB - Pivotal brain functions, such as neurotransmission, cognition, and memory, decline with advancing age and, especially, in neurodegenerative conditions associated with aging, such as Alzheimer's disease (AD). Yet, deterioration in structure and function of the nervous system during aging or in AD is not uniform throughout the brain. Selective neuronal vulnerability (SNV) is a general but sometimes overlooked characteristic of brain aging and AD. There is little known at the molecular level to account for the phenomenon of SNV. Functional genomic analyses, through unbiased whole genome expression studies, could lead to new insights into a complex process such as SNV. Genomic data generated using both human brain tissue and brains from animal models of aging and AD were analyzed in this review. Convergent trends that have emerged from these data sets were considered in identifying possible molecular and cellular pathways involved in SNV. It appears that during normal brain aging and in AD, neurons vulnerable to injury or cell death are characterized by significant decreases in the expression of genes related to mitochondrial metabolism and energy production. In AD, vulnerable neurons also exhibit down-regulation of genes related to synaptic neurotransmission and vesicular transport, cytoskeletal structure and function, and neurotrophic factor activity. A prominent category of genes that are up regulated in AD are those related to inflammatory response and some components of calcium signaling. These genomic differences between sensitive and resistant neurons can now be used to explore the molecular underpinnings of previously suggested mechanisms of cell injury in aging and AD. PMID- 21629440 TI - Mesenchymal stem cells for treatment of CNS injury. AB - Brain and spinal cord injuries present significant therapeutic challenges. The treatments available for these conditions are largely ineffective, partly due to limitations in directly targeting the therapeutic agents to sites of pathology within the central nervous system (CNS). The use of stem cells to treat these conditions presents a novel therapeutic strategy. A variety of stem cell treatments have been examined in animal models of CNS trauma. Many of these studies have used stem cells as a cell-replacement strategy. These investigations have also highlighted the significant limitations of this approach. Another potential strategy for stem cell therapy utilises stem cells as a delivery mechanism for therapeutic molecules. This review surveys the literature relevant to the potential of mesenchymal stem cells for delivery of therapeutic agents in CNS trauma in humans. PMID- 21629441 TI - Pharmacological neuroprotection after perinatal hypoxic-ischemic brain injury. AB - Perinatal hypoxia-ischemia (HI) is an important cause of neonatal brain injury. Recent progress in the search for neuroprotective compounds has provided us with several promising drugs to reduce perinatal HI-induced brain injury. In the early stage (first 6 hours after birth) therapies are concentrated on prevention of the production of reactive oxygen species or free radicals (xanthine-oxidase-, nitric oxide synthase-, and prostaglandin inhibition), anti-inflammatory effects (erythropoietin, melatonin, Xenon) and anti-apoptotic interventions (nuclear factor kappa B- and c-jun N-terminal kinase inhibition); in a later stage stimulation of neurotrophic properties in the neonatal brain (erythropoietin, growth factors) can be targeted to promote neuronal and oligodendrocyte regeneration. Combination of pharmacological means of treatment with moderate hypothermia, which is accepted now as a meaningful therapy, is probably the next step in clinical treatment to fight post-asphyxial brain damage. Further studies should be directed at a more rational use of therapies by determining the optimal time and dose to inhibit the different potentially destructive molecular pathways or to enhance endogenous repair while at the same time avoiding adverse effects of the drugs used. PMID- 21629442 TI - Neuro-psychopharmacogenetics and Neurological Antecedents of Posttraumatic Stress Disorder: Unlocking the Mysteries of Resilience and Vulnerability. AB - BACKGROUND AND HYPOTHESIS: Although the biological underpinnings of immediate and protracted trauma-related responses are extremely complex, 40 years of research on humans and other mammals have demonstrated that trauma (particularly trauma early in the life cycle) has long-term effects on neurochemical responses to stressful events. These effects include the magnitude of the catecholamine response and the duration and extent of the cortisol response. In addition, a number of other biological systems are involved, including mesolimbic brain structures and various neurotransmitters. An understanding of the many genetic and environmental interactions contributing to stress-related responses will provide a diagnostic and treatment map, which will illuminate the vulnerability and resilience of individuals to Posttraumatic Stress Disorder (PTSD). PROPOSAL AND CONCLUSIONS: We propose that successful treatment of PTSD will involve preliminary genetic testing for specific polymorphisms. Early detection is especially important, because early treatment can improve outcome. When genetic testing reveals deficiencies, vulnerable individuals can be recommended for treatment with "body friendly" pharmacologic substances and/or nutrients. Results of our research suggest the following genes should be tested: serotoninergic, dopaminergic (DRD2, DAT, DBH), glucocorticoid, GABAergic (GABRB), apolipoprotein systems (APOE2), brain-derived neurotrophic factor, Monamine B, CNR1, Myo6, CRF-1 and CRF-2 receptors, and neuropeptide Y (NPY). Treatment in part should be developed that would up-regulate the expression of these genes to bring about a feeling of well being as well as a reduction in the frequency and intensity of the symptoms of PTSD. PMID- 21629443 TI - Use of antiepileptic drugs for hyperkinetic movement disorders. AB - Many studies investigated the use of antiepileptic drugs (AEDs) in several neurological diseases other than epilepsy. These neurological disorders, usually, involve neuronal excitability through the modulating of ion channels, receptors and intracellular signaling pathways, and are the targets of the AEDs. This article provides a review of the clinical efficacy of both conventional and newer AEDs in hyperkinetic movement disorders. Some of these indications for AEDs have been established, while others are under investigation. The modulation of GABAergic transmission may explain the neuronal hyper-excitability that underlies some forms of hyperkinetic movement disorders. So, AEDs able to increase GABAergic neurotransmission may play a role in hyperkinetic movement disorders treatment. Therefore, AEDs could represent a useful therapeutic option in the management of hyperkinetic movement disorders where the available treatments are ineffective. PMID- 21629444 TI - The neuropharmacology of implicit learning. AB - Two decades of pharmacologic research on the human capacity to implicitly acquire knowledge as well as cognitive skills and procedures have yielded surprisingly few conclusive insights. We review the empirical literature of the neuropharmacology of implicit learning. We evaluate the findings in the context of relevant computational models related to neurotransmittors such as dopamine, serotonin, acetylcholine and noradrenalin. These include models for reinforcement learning, sequence production, and categorization. We conclude, based on the reviewed literature, that one can predict improved implicit acquisition by moderately elevated dopamine levels and impaired implicit acquisition by moderately decreased dopamine levels. These effects are most prominent in the dorsal striatum. This is supported by a range of behavioral tasks in the empirical literature. Similar predictions can be made for serotonin, although there is yet a lack of support in the literature for serotonin involvement in classical implicit learning tasks. There is currently a lack of evidence for a role of the noradrenergic and cholinergic systems in implicit and related forms of learning. GABA modulators, including benzodiazepines, seem to affect implicit learning in a complex manner and further research is needed. Finally, we identify allosteric AMPA receptors modulators as a potentially interesting target for future investigation of the neuropharmacology of procedural and implicit learning. PMID- 21629445 TI - Gene expression profiling in rodent models for schizophrenia. AB - The complex neurodevelopmental disorder schizophrenia is thought to be induced by an interaction between predisposing genes and environmental stressors. In order to get a better insight into the aetiology of this complex disorder, animal models have been developed. In this review, we summarize mRNA expression profiling studies on neurodevelopmental, pharmacological and genetic animal models for schizophrenia. We discuss parallels and contradictions among these studies, and propose strategies for future research. PMID- 21629446 TI - Glutamatergic targets for enhancing extinction learning in drug addiction. AB - The persistence of the motivational salience of drug-related environmental cues and contexts is one of the most problematic obstacles to successful treatment of drug addiction. Behavioral approaches to extinguishing the salience of drug associated cues, such as cue exposure therapy, have generally produced disappointing results which have been attributed to, among other things, the context specificity of extinction and inadequate consolidation of extinction learning. Extinction of any behavior or conditioned response is a process of new and active learning, and increasing evidence suggests that glutamatergic neurotransmission, a key component of the neural plasticity that underlies normal learning and memory, is also involved in extinction learning. This review will summarize findings from both animal and human studies that suggest that pharmacological enhancement of glutamatergic neurotransmission facilitates extinction learning in the context of drug addiction. Pharmacological agents that have shown potential efficacy include NMDA partial agonists, mGluR5 receptor positive allosteric modulators, inhibitors of the GlyT1 glycine transporter, AMPA receptor potentiators, and activators of the cystine-glutamate exchanger. These classes of cognition-enhancing compounds could potentially serve as novel pharmacological adjuncts to cue exposure therapy to increase success rates in attenuating cue-induced drug craving and relapse. PMID- 21629447 TI - "Epileptic encephalopathy" of infancy and childhood: electro-clinical pictures and recent understandings. AB - There is growing interest in the diagnosis of cognitive impairment among children with epilepsy. It is well known that status of seizures control has to be carefully investigated because it can be sufficient "per se" to cause progressive mental deterioration conditions. Subclinical electroencephalographic discharges may have subtle effects on cognition, learning and sleep patterns, even in the absence of clinical or sub-clinical seizures. In this respect, electroencephalographic monitoring (long-term and nocturnal recording) and in particular an all night video-polysomnography (V-NPSG) record can be crucial to detect the presence of unrecognized seizures and/or an inter-ictal nocturnal EEG discharge increasing. Epileptic encephalopathies (EE) are a group of conditions in which the higher cognitive functions are deteriorate as a consequence of epileptic activity, which, in fact, consists of frequent seizures and/or florid and prolonged interictal paroxysmal discharges, focal or generalized. AEDs represent the first line in opposing the burden of both, the poor seizures control and the poor interictal discharges control, in the cognitive deterioration of EE affected children. Thus, to improve the long-term cognitive/behavioural prognosis in these refractory epileptic children, it should be taken into account both a good seizures control and a strict sleep control, choosing carefully antiepileptic drugs which are able to control not only seizures clinically recognizable but even the EEG discharges onset and its increasing and spreading during sleep. Here, we review the efficacy and safety of the newer AEDs that, to date, are used in the treatment of EE in infancy and childhood. PMID- 21629449 TI - Conventional and contemporary luting cements: an overview. AB - Long-term clinical success of fixed prosthodontic restorations is influenced by many factors, one important factor being the selection of an appropriate luting agent. No single luting agent is capable of meeting all the stringent requirements, which is one reason why there is such a wide choice of luting agents currently available from conventional water-based to contemporary adhesive resin cements. Introduction of adhesive resin systems has completely changed the face of fixed prosthodontic practice leading to an increased use of bonded all ceramic crowns and resin-retained fixed partial dentures. This article makes an effort to review various conventional and contemporary luting agents, their properties & associated clinical implications thereby trying to help the clinician select an appropriate luting agent for a given clinical situation. PMID- 21629450 TI - An in vitro study to evaluate the retention of complete crowns prepared with five different tapers and luted with two different cements. AB - This study includes comparatively evaluating and drawing general conclusion about the best advisable taper and cement for maximal retention. Eighty extracted human maxillary premolar teeth with sound surfaces were selected using dial vernier caliper. Selected teeth were randomly divided into five different taper groups (0 degrees , 3 degrees , 6 degrees , 9 degrees and 12 degrees ). The crown preparations with different tapers respectively were achieved by graduated customized device. Preparations were verified with special set up. Crowns were cast with Co-Cr alloy; metal copings were luted with glass ionomer and zinc phosphate cement. Retention was measured (MPa) by separating the metal crowns from the teeth under tension on a universal testing machine, data was recorded and statistically analyzed. Glass ionomer cemented 0 degrees and 12 degrees taper group showed increase in retentive strength (p = 0.003 hs), when compared to zinc phosphate cement. 9 degrees and 12 degrees group showed decreased retentive strength (p = 0.001 vhs) when compared with 0 degrees taper group. No significant difference found between 0 degrees and 3 degrees and 6 degrees group. The choice of cement for crowns prepared within this ideal range (0 degrees -6 degrees taper) might be of limited clinical significance. 3 degrees and 6 degrees taper with zinc phosphate or glass ionomer cement shown to be ideal for maximum retention. PMID- 21629451 TI - Prevalence of signs of temporomandibular joint dysfunction in asymptomatic edentulous subjects: a cross-sectional study. AB - Patients having complete dentures with reduced vertical dimension generally do not manifest Temporomandibular Joint problems. It is not understood as to why the closure of jaws in dentulous individuals can predispose to Temporomandibular Joint problems, while the same etiology in edentulous subjects does not cause any concern. This study was planned to find out the prevalence of various Temporomandibular Joint dysfunction signs in subjects who were edentulous for a period of 6 months to 2 year. The various signs were obtained from a population of 100 healthy asymptomatic edentulous subjects by a questionnaire and then were clinically examined for the presence or absence of signs of Temporomandibular Joint dysfunction. 59% of the subjects exhibited one or more signs of Temporomandibular Joint dysfunction, 41% of the subjects did not show any signs of Temporomandibular Joint dysfunction. 56.6% of males reported signs of Temporomandibular Joint dysfunction when compared with females which was 62.5%. 43.3% of males did not show any signs of Temporomandibular Joint dysfunction when compared with those of the females which was 37.5%. The number of subjects who showed two signs was 29%, subjects who had only one sign was 25%. It was found that dysfunction was prevalent among both men and women in more than half of the asymptomatic subjects examined. 59% had one or as many as three signs of Temporomandibular dysfunction. The most commonly seen Joint dysfunction was the joint sounds which was 47% PMID- 21629448 TI - An Update on GABArho Receptors. AB - The present review discusses the functional and molecular diversity of GABArho receptors. These receptors were originally described in the mammalian retina, and their functional role in the visual pathway has been recently elucidated; however new studies on their distribution in the brain and spinal cord have revealed that they are more spread than originally thought, and thus it will be important to determine their physiological contribution to the GABAergic transmission in other areas of the central nervous system. In addition, molecular modeling has revealed peculiar traits of these receptors that have impacted on the interpretations of the latest pharmacolgical and biophysical findings. Finally, sequencing of several vertebrate genomes has permitted a comparative analysis of the organization of the GABArho genes. PMID- 21629452 TI - A simplified approach for recording neutral zone. AB - Neutral zone technique is long being used for the management of severely resorbed mandibular ridges. Various materials are used in recording neutral zone, which have their own advantages and disadvantages. This article discusses the use of Polyether impression material which is simpler and more practical. PMID- 21629453 TI - A systematic review of impression technique for conventional complete denture. AB - The importance of an in depth review of impression making for complete dentures lies in the assessment of the historical value of all the factors related to physical, biologic and behavioral areas and the time in which they were discussed and taught as well. This review documents the historical development of knowledge associated with scientific advancement from 1845 to the present year, i.e. 2009 related to impression procedures in conventional complete denture prosthesis. Search for articles was done through electronic media the Pubmed. PMID- 21629454 TI - A comparative Study of the Masticatory Efficiency in Complete Dentures Using Acrylic and Metal Occlusal Posterior Teeth-Photocolorimetric Analysis. AB - Masticatory efficiency is always compromised in complete denture patients. The denture teeth with compromised occlusal anatomy and material further compound the inherent limitations of complete dentures. To compare the masticatory efficiency of patients with complete denture using acrylic and metal occlusal posterior teeth. Ten edentulous patients with well-formed ridges were selected. Complete dentures using acrylic and metal occlusal posterior teeth were fabricated. The masticatory efficiency was calculated using photo colorimeter. Complete dentures with 20 degrees acrylic resin posterior teeth were fabricated for ten edentulous patients and masticatory efficiency was measured using raw beetroot as test food on a photocolorimeter after 10, 15, 20 strokes. The acrylic resin posterior teeth were replaced with metal occlusal posterior teeth on the same denture base. The masticatory efficiency was measured by the same method. The mean average percentage of masticatory efficiency with acrylic resin teeth and metal occlusal teeth were first calculated and statistical analysis was done using students unpaired t test. The average masticatory efficiency with acrylic teeth was 46.46% and metal occlusal teeth was 65.62, 't' = 5.38. The difference between the mean average masticatory performance percentage with acrylic teeth and metal occlusal teeth was significant at 1% i.e. P < 0.01. Within the limitation of this study the average percentage value of masticatory efficiency improves considerably when metal occlusal teeth replace normal acrylic teeth without any alteration in position or relation. PMID- 21629455 TI - Prefabricated stock trays for impression of auricular region. AB - The conventional methods of impression making for maxillofacial defects are cumbersome and time consuming for both patient and operator. This study focuses upon standardizing and simplifying the impression making methodology for auricular prosthesis with the help of prefabricated stock trays for auricular region. The stock trays were designed on positive replicas of anatomical structures, broadly divided into long and narrow, short and broad and long and broad ear. For each stock tray, impressions of auricle, of patients of different morphology were made with plastic funnels of different shape and size ensuring at least 6 mm of space between the anatomical part and inner surface of funnel and master cast was obtained. Subsequent adaptation of wax was done and fabrications of stock stainless steel trays were done. A standardized stock tray for making of auricular impressions was developed. From this innovative technical procedure it is possible to get an accurate impression of auricular defects now by the use of prefabricated stock trays rather than the cumbersome conventional method. PMID- 21629456 TI - Correlation of nasal width to inter-canine distance in various arch forms. AB - Selection of teeth for dentures to provide a aesthetically pleasing appearance is a challenge often encountered. Various anthropometric measurements have been utilised traditionally, which is subjective to each practitioner. Interalar width is one of the routinely used method in the selection of upper anterior teeth. It is based on the premise that the parallel line drawn from the alae of the nose touches the tip of the canine. However this may not hold good for all type of arch forms. This paper finds a correlation between the alar of the nose and the tip of the canine for the various arch forms. PMID- 21629457 TI - Modification of esthetics using a combined orthodontic and a prosthodontic approach: a case report. AB - A patient sometimes requires a multidisciplinary approach to correct the esthetics and to improve the occlusion. This case report describes the management of an adult female patient with a convex profile, proclined upper and lower anterior teeth, missing upper left lateral incisor and peg shaped upper right lateral incisor tooth through orthodontic and prosthodontic treatment. PMID- 21629458 TI - A novel method for retention of an orbital prosthesis in a case with continuous maxillary and orbital defect. AB - Exenteration of the orbital contents as well as removal of a part of maxilla with an ablative surgery for the removal of a malignant tumor can severely affect a person in terms of function, esthetics and psychological trauma. A well-retained, user-friendly, removable maxillofacial prosthesis is the key to successful prosthetic rehabilitation in such cases. Various retentive techniques include using spectacle frame, conformers, adhesives, osseointegrated implants, magnets or buttons. This paper describes a novel technique for retention of silicone orbital prosthesis using acrylic resin base attached to maxillary obturator with the help of pin and socket of an electric plug wherein the latter is attached to the eye prosthesis with the help of a mechanical undercut. This helps in better retention of both the prostheses. PMID- 21629459 TI - Prosthodontic management of a child with ectodermal dysplasia: a case report. AB - Ectodermal dysplasia is a rare congenital disease that affects several ectodermal structures. Children with ectodermal dysplasia may have various manifestation of the disease that differ in severity and may or may not involve teeth, skin, hair, nails, sweat glands and sebaceous glands. The most common form of the ectodermal dysplasia syndrome is hypohidrotic ectodermal dysplasia and is usually inherited as an X-linked recessive trait. Female carriers may have a variable degree of clinical manifestations. This case report discusses the management of a 5-year old girl with ectodermal dysplasia. Clinical management consisted of fabricating upper and lower dentures to help in psychosocial development and to restore the vertical dimension, esthetics and functioning of the stomatognathic system. PMID- 21629460 TI - The importance of diagnostic test parameters in the interpretation of clinical test findings: The Prone Hip Extension Test as an example. AB - The use of diagnostic tests is a crucial aspect of clinical practice since they assist clinicians in establishing whether a patient has or does not have a particular condition. In order for any clinical test to be used most appropriately, it is essential that several parameters be established regarding the test and that these are made known to clinicians to inform their clinical decision making. These include the test's sensitivity, specificity, predictive values, and likelihood ratios. This article reviews their importance as well as provides an illustrative example that highlights how knowledge of the parameters for a given test allows clinicians to better interpret their test findings in practice. PMID- 21629461 TI - A narrative review of the published chiropractic literature regarding older patients from 2001-2010. AB - INTRODUCTION: The purpose of this article was to perform a narrative review of the chiropractic literature regarding older patients between 2001 and 2010. METHODS: A three step search strategy of the literature involved electronic searching, hand searching and reference tracking. RESULTS: One hundred and eighty eight articles germane to chiropractic geriatric practice and education were retrieved. DISCUSSION: Compared to the review of the literature conducted prior to 2000, the number of references on chiropractic geriatric education increased from 3 to 11, the number of demographic studies increased from 9 to 18, the number of case reports increased from 25 to 83, the number of clinical trials increased from 4 to 21 (only two RCTs found) and the number of references on clinical guidelines and general clinical information increased from 18 to 55. CONCLUSION: This review found 188 retrievable articles available to practitioners to effectively care plan for their older patients, a better than three fold increase in the number of references found during a similar review conducted at the end of the previous decade. However, there is clearly a gap in the evidence base of chiropractic geriatric care, particularly the under-representation of clinical trials of all kinds involving older chiropractic patients. PMID- 21629462 TI - Impairment assessment of lateral epicondylitis through electromyography and dynamometry. AB - OBJECTIVES: To investigate changes in muscular activity and strength of subjects diagnosed with lateral epicondylitis (LE). To assess the appropriateness of these measures in the patient's follow-up. METHODS: Twenty-four subjects (11 men and 13 women) with LE, were evaluated at baseline and after 5 weeks of an experimental treatment. Measurements included: the (1) pain-free grip (PFG), (2) maximal isometric strength, (3) surface electromyography (EMG) of forearm muscle (healthy and affected), (4) a visual analogue scale (VAS), and (5) the Patient Rated Tennis Elbow Evaluation (PRTEE) (Canadian-French version). RESULTS: All subjects showed improvement in VAS and PRTEE. The maximal isometric strength during flexion and extension of the wrist and the EMG analysis failed to discriminate the affected from the healthy elbow during the initial assessment. Only the PFG measured with the elbow in extension could discriminate elbows with LE from the healthy ones. CONCLUSIONS: The use of the PFG with the elbow in extension seems to be the most indicated strength measurement to monitor the recovery of patients with LE. The EMG acquisition protocol used in this research was not adequate to monitor effectively the recovery of LE. PMID- 21629464 TI - Two cases of medial knee pain involving the medial coronary ligament in adolescents treated with conservative rehabilitation therapy. AB - OBJECTIVE: This case study chronicled the assessment, treatment and management of two adolescent patients presenting with acute knee pain, diagnosed as medial meniscus tear, with or without a medial collateral ligament sprain, with coronary ligament involvement. CASES: Patient 1: A 16 year old male football player presented with right medial knee pain of 2 days duration after having been tackled during practice from the left side. Patient 2: A 16 year old female presented with right medial knee pain that began 1 week prior to presentation after a fall down the stairs. TREATMENT: Treatment was initiated in both cases using inflammatory control techniques of icing and fascial stripping and progressed to rehabilitative exercises including VMO (vastus medialis oblique) exercises and squatting exercises to strengthen the quadriceps femoris musculature and proprioceptive exercise. Rehabilitation occurred over a four week duration in both cases with progression of exercises on an individual basis. Both cases resolved within four weeks and return to normal activities resumed at the three week mark including a return to play in patient 1. Both patients reported complete resolution of symptoms at the four week mark with no recurrence on follow up a number of months later. SUMMARY: Conservative management, including icing, fascial stripping, and rehabilitative exercises may be beneficial in the treatment of medial meniscus tears with coronary ligament involvement in adolescents. PMID- 21629463 TI - Attitudes of clinicians at the Canadian Memorial Chiropractic College towards the chiropractic management of non-musculoskeletal conditions. AB - OBJECTIVE: The objective of this study was to determine the attitudes of clinical faculty during the 2009-2010 academic year at the Canadian Memorial Chiropractic College towards the treatment of various non-musculoskeletal disorders. METHODS: A confidential survey was distributed to the clinical faculty via email. It consisted of several questions polling the demographic of the respondent such as years in clinical practice, and a list of 29 non-musculoskeletal conditions. Clinicians were asked to indicate their opinions on each condition on rating scale ranging from strongly agree to strongly disagree. RESULTS: Twenty of 22 clinicians responded. The conditions garnering the greatest positive ratings include: asthma, constipation, chronic pelvic pain, dysmenorrhea, infantile colic, and vertigo. The options regarding vertigo and asthma, while demonstrating an overall positive attitude towards the benefits of chiropractic care, were stratified amongst clinicians with varying years in clinical practice. CONCLUSION: This study suggests clinicians at this college are moderately open towards the chiropractic treatment of some non-musculoskeletal disorders. PMID- 21629465 TI - The West family chiropractic dynasty: celebrating a century of accomplishment in Canada: Part II: Samson J. West, David I. West, Neil A. West, Megan L. West, R. Ian Buchanan and James L. West. AB - This historical paper documents the unbroken legacy of the West family of chiropractors which has flourished in Canada for over 100 years. Part I, unearthed the origins, development and careers of Archibald West, the founder of this dynasty, his son Samuel and grandson Stephen. Part II, delves into the life of Archie's brother Samson, and Samson's chiropractic progeny: grandsons David and Neil, and great granddaughter Megan. Then it goes back to look at Stephen West's nephew, R. Ian Buchanan and ends with a descendant of another branch of the family tree, James L. West. PMID- 21629467 TI - Treating a disease vs. understanding a disease. PMID- 21629468 TI - Pedicle screw instrumentation for adolescent idiopathic scoliosis: the insertion technique, the fusion levels and direct vertebral rotation. AB - The pedicle is a power nucleus of the vertebra and offers a secure grip of all 3 columns. Pedicle screw instrumentation has advantages of rigid fixation with improved three-dimensional (3D) correction and it is accepted as a reliable method with a high margin of safety. Accurate placement of the pedicle screws is important to reduce possible irreversible complications. Many methods of screw insertion have been reported. The author has been using the K-wire method coupled with the intraoperative single posteroanterior and lateral radiographs, which is the most safe, accurate and fast method. Identification of the curve patterns and determining the fusion levels are very important. The ideal classification of adolescent idiopathic scoliosis should address the all patterns, predict the extent of accurate fusion and have good inter/intraobserver reliability. My classification system matches with the ideal classification system, and it is simple and easy to learn; and my classification system has only 4 structural curve patterns and each curve has 2 types. Scoliosis is a 3D deformity; the coronal and sagittal curves can be corrected with rod rotation, and rotational deformity has to be corrected with direct vertebral rotation (DVR). Rod derotation and DVR are true methods of 3D deformity correction with shorter fusion and improved correction of both the fused and unfused curves, and this is accomplished using pedicle screw fixation. The direction of DVR is very important and it should be opposite to the direction of the rotational deformity of the vertebra. A rigid rod has to be used to prevent rod bend-out during the derotation and DVR. PMID- 21629469 TI - Treatment of periprosthetic femoral fractures in hip arthroplasty. AB - BACKGROUND: We analyzed the radiological and clinical results of our study subjects according to the management algorithm of the Vancouver classification system for the treatment of periprosthetic femoral fractures in hip arthroplasty. METHODS: We retrospectively reviewed 18 hips with postoperative periprosthetic femoral fractures. The average follow-up was 49 months. The fracture type was determined based on the Vancouver classification system. The management algorithm of the Vancouver classification system was generally applied, but it was modified in some cases according to the surgeon's decision. At the final follow-up, we assessed the radiological results using Beals and Tower's criteria. The functional results were also evaluated by calculating the Harris hip scores. RESULTS: Seventeen of 18 cases (94.4%) achieved primary union at an average of 25.5 weeks. The mean Harris hip score was 92. There was 1 case of nonunion, which was a type C fracture after cemented total hip arthroplasty, and this required a strut allograft. Subsidence was noted in 1 case, but the fracture was united despite the subsidence. There was no other complication. CONCLUSIONS: Although we somewhat veered out of the management algorithm of the Vancouver classification system, the customized treatment, with considering the stability of the femoral stem and the configuration of the fracture, showed favorable overall results. PMID- 21629470 TI - Additional fixations for sliding hip screws in treating unstable pertrochanteric femoral fractures (AO Type 31-A2): short-term clinical results. AB - BACKGROUND: To evaluate the utility of additional fixation methods and to suggest a method of reduction in the treatment of unstable pertrochanteric femur fractures with a sliding hip screw (SHS). METHODS: A retrospective study was performed on thirty patients with unstable pertrochanteric femur fractures, who were operated on with a SHS between September 2004 and September 2009 and were followed up for at least 6 months. The additional fixation devices were as follows; antirotation screw (21 cases), fixation of displaced fractures of the posteromedial bone fragment (cerclage wiring, 21 cases and screw, 2 cases) and trochanter stabilizing plate (27 cases). Clinically, the Palmer's mobility score and Jensen's social function group were used. Radiologically, alignment and displacement were observed. The tip-apex distance (TAD) and sliding of the lag screw were measured, and the position of the lag screw within the femoral head was also examined. RESULTS: The mean age at the time of surgery was 76 years (range, 56 to 89 years) and the average follow-up period was 25 months (range, 6 to 48 months). At the last follow-up, the average mobility and social function score was 6.2 (+/- 3.5) and 2.3 (+/- 1.5). Postoperatively, the alignment and displacement indices were adequate in almost all the cases. The mean amount of lag screw sliding and the mean TAD was 5.1 mm (range, 2 to 16 mm) and 6 mm (range, 3 to 11 mm) respectively. The lag screws were located in the center center zone in 21 cases. The average period to union was 18.7 weeks without any cases of nonunion or malunion. Mechanical failure was noted in one case with breakage of the lag screw and clinical failure was noted in another case with persistent hip pain related to excessive sliding (16 mm). CONCLUSIONS: With additional fixations, the unstable pertrochanteric femur fractures could be well stabilized by SHS until bone union. PMID- 21629471 TI - Reconstruction of acetabular posterior wall fractures. AB - BACKGROUND: The results after acetabular fracture are primarily related to the quality of the articular reduction. We evaluated the results of internal fixation of posterior wall fractures with using three-step reconstruction. METHODS: Thirty three patients (mean age at the time of injury, 47.9 years; 28 males and 5 females) were followed for a minimum of 2 years after surgery. The three-step reconstruction included 1) preservation of soft tissues and reduction of the marginally impacted osteochondral (articular) fragments using screws, 2) filling the impacted cancellous void with a bone graft, and 3) reinforcement with buttress-plating. Clinical evaluation was done according to the criteria of D'aubigne and Postel, while the radiological criteria were those of Matta. The associated injuries and complications were evaluated. RESULTS: The clinical results were excellent in 15 (45.5%) patients and they were good in 5 (15.2%), (i.e., satisfactory in 60.7%), while the radiologic results were excellent in 10 (30.3%) and good in 14 (42.4%) (satisfactory in 72.7%). Heterotopic ossification was common, but this did not require excision, even without prophylactic treatment with indomethacin. Deep infection was the worst complication and this was accompanied by a poor outcome. CONCLUSIONS: This study confirms that three step reconstruction facilitates accurate and firm reduction of displaced posterior wall fractures of the acetabulum. Therefore, we anticipate less long term arthrosis in the patients treated this way. PMID- 21629472 TI - The correlations of the radiological parameters of hip dysplasia and proximal femoral deformity in clinically normal hips of a Korean population. AB - BACKGROUND: The aim of this study was to answer the following two questions: 1) Do the radiological parameters of dysplasia have significant correlations between themselves or with the parameters of the proximal femoral deformity and vice versa? 2) Do the physical parameters have a significant correlation with the radiological parameters of hip dysplasia and proximal femoral deformity? METHODS: Four hundred and twenty eight consecutive patients with no clinical evidence of hip osteoarthritis and who underwent pelvic radiography in the supine position for hip contusion or a routine health check were analyzed for the relationships between the center-edge (CE) angle, acetabular depth, acetabular angle, the head neck ratio and the neck-shaft angle as well as the relationships of the above mentioned variables with age, gender, body height and the body mass index. RESULTS: The CE angle, acetabular depth and acetabular angle showed a strong correlation with each other. The neck-shaft angle and the head-neck ratio showed no correlation with each other or with the CE angle, acetabular depth and acetabular angle. Age was positively associated with the CE angle, and inversely associated with the acetabular depth or acetabular angle. Male gender was significantly associated with the increased neck-shaft angle, and inversely associated with the head-neck ratio. CONCLUSIONS: The radiological parameters of hip dysplasia are all strongly, if not perfectly, inter-correlated. Age was associated with the radiological parameters of hip dysplasia whereas gender was associated with the radiological parameters of a proximal femoral deformity. PMID- 21629473 TI - Characteristics of radial tears in the posterior horn of the medial meniscus compared to horizontal tears. AB - BACKGROUND: The clinical and radiologic features of radial tears of the medial meniscus posterior horn were compared with those of horizontal tears. METHODS: From January 2007 to December 2008, 387 consecutive cases of medial meniscal tears were treated arthroscopically. Among these, 91 were radial tears in the medial meniscus posterior horn, and 95 were horizontal tears in the posterior segment of the medial meniscus. The patients' data (age, gender, duration of symptom, body mass index, and injury history), radiographic findings (Kellgren and Lawrence score, posterior tibial slope, and femorotibial angle), and chondral lesions were recorded. RESULTS: The patient factors of age, gender, and body mass index were related to radial tears of the medial meniscus posterior horn. Radial tears were significantly correlated with Kellgren and Lawrence score, varus alignment, posterior tibial slope, and severity of the chondral lesion. CONCLUSIONS: Radial tears of the medial meniscus posterior horn are a unique clinical entity that are associated with older age, females and obesity, and are strongly associated with an increased incidence and severity of cartilage degeneration compared to horizontal tears. PMID- 21629474 TI - Surgical management of ipsilateral fracture of the femur and tibia in adults (the floating knee): postoperative clinical, radiological, and functional outcomes. AB - BACKGROUND: This study evaluated the outcomes of surgical management of ipsilateral femoral and tibial fractures in adults. METHODS: Fifteen patients (13 men, 2 women; mean age, 34.8 years; range, 18 to 65 years) were enrolled in this study. The fractures types were classified according to the classification by Fraser et al. as follows: type I (5), type IIa (3), IIb (4), IIc (3). Femur fractures were treated using locked intramedullary nails, plate-screws, or dynamic condylar screws, and tibia fractures were treated with an external fixator (in open fractures), or plate-screws, and locked intramedullary nailing. The mean follow-up duration was 2.2 years (range, 1.3 to 4 years). RESULTS: The extent of bony union according to the Karlstrom criteria was as follows: excellent, 8; good, 4; acceptable, 2; poor, 1. CONCLUSIONS: The associated injuries and type of fracture (open, intra-articular, comminution) are prognostic factors in a floating knee. The best management of the associated injuries for good final outcome involves intramedullary nailing of both the fractures and postoperative rehabilitation. PMID- 21629475 TI - Fractures of the proximal fifth metatarsal: percutaneous bicortical fixation. AB - BACKGROUND: Displaced intraarticular zone I and displaced zone II fractures of the proximal fifth metatarsal bone are frequently complicated by delayed nonunion due to a vascular watershed. Many complications have been reported with the commonly used intramedullary screw fixation for these fractures. The optimal surgical procedure for these fractures has not been determined. All these observations led us to evaluate the effectiveness of percutaneous bicortical screw fixation for treating these fractures. METHODS: Twenty-three fractures were operatively treated by bicortical screw fixation. All the fractures were evaluated both clinically and radiologically for the healing. All the patients were followed at 2 or 3 week intervals till fracture union. The patients were followed for an average of 22.5 months. RESULTS: Twenty-three fractures healed uneventfully following bicortical fixation, with a mean healing time of 6.3 weeks (range, 4 to 10 weeks). The average American Orthopaedic Foot & Ankle Society (AOFAS) score was 94 (range, 90 to 99). All the patients reported no pain at rest or during athletic activity. We removed the implant in all cases at a mean of 23.2 weeks (range, 18 to 32 weeks). There was no refracture in any of our cases. CONCLUSIONS: The current study shows the effectiveness of bicortical screw fixation for displaced intraarticular zone I fractures and displaced zone II fractures. We recommend it as one of the useful techniques for fixation of displaced zone I and II fractures. PMID- 21629476 TI - Responsiveness of the Korean version of the disabilities of the arm, shoulder and hand questionnaire (K-DASH) after carpal tunnel release. AB - BACKGROUND: The Korean version of the Disability of the Arm, Shoulder and Hand Questionnaire (K-DASH) was recently validated, but its responsiveness, which is the degree to which an instrument is sensitive to change, has not been thoroughly evaluated in a specific condition in Koreans. We evaluated the responsiveness of the K-DASH in a homogenous cohort of patients with carpal tunnel syndrome (CTS) and we compared it with that of the disease-specific Carpal Tunnel Questionnaire (CTQ). METHODS: Fifty-six patients with CTS prospectively completed the K-DASH and CTQ before and 6 months after surgery. The responsiveness statistics were assessed for both the K-DASH and CTQ by using the standardized response mean (SRM) and the effect size (ES). Pearson correlation coefficients were calculated between the K-DASH and CTQ. RESULTS: The SRM and ES of the K-DASH were all 0.8. The SRM of the symptom and function part of the CTQ was 1.5 and 1.1, and the ES was 1.5 and 1.1, respectively. The K-DASH had moderate correlations with the symptom and function parts of the CTQs, but the postoperative K-DASH had a weak correlation with the symptom part of the postoperative CTQ. CONCLUSIONS: The K DASH was found to have a large degree of responsiveness (SRM, ES >= 0.8) after carpal tunnel release in Korean patients with CTS, which is comparable to the other language versions of the DASH. Although it was less responsive than the CTQ, which is disease-specific, the region-specific K-DASH can be used as an effective outcome measurement tool for CTS, and especially for research that compares CTS with other upper limb conditions. PMID- 21629477 TI - Ganglionectomy without repairing the bursal defect: long-term results in a series of 124 wrist ganglia. AB - BACKGROUND: Some surgeons consider the abscission of a part of the articular bursa around the point of the input of ganglion's nape (average 1-2 cm diameter) to be very important with excellent results. However, a literature search revealed disagreement as to whether it is essential to repair a bursa defect. This study examined the effectiveness of this method without repairing the articular defect. An attempt was made to identify the anatomical origin of wrist ganglia during the surgical procedure. METHODS: This study evaluated 124 wrist ganglia that had been treated surgically during 2004-2009 using this technique and without repairing the bursa defect (1-2 cm in diameter). The variables studied were age, gender, time from the occurrence till abscission of the ganglia, former surgical interventions, preoperative and postoperative pain, insertion of the ganglion's nape and complications. Sixty-six patients with a mean follow-up of 42 months and minimum 12 months were examined. RESULTS: At the time of the follow-up, 80.3% had no pain whereas 92.2% showed a remarkable improvement. Seven cases of recurrence (10.6%) were found 2 to 85 months after surgery, of which most appeared during the first year (71.4%). It is important to mention that the majority of the dorsal ganglia (42.8%) originated from the capitate-lunate joint. None of the patients presented with scapholunate or other instability. CONCLUSIONS: This surgical method is a simple and safe with excellent long-term results and a lower recurrence rate compared to other surgical approaches. Overall, repair of the articular bursa is unnecessary. PMID- 21629478 TI - The natural history of pediatric trigger thumb: a study with a minimum of five years follow-up. AB - BACKGROUND: Pediatric trigger thumb is due to deformed flexion of the interphalangeal joint. We previously reported that pediatric trigger thumb can spontaneously resolve in > 60% of patients at the median follow-up of 48 months. The purpose of this study was to determine whether there were any more cases of resolution with a follow-up of more than 5 years and whether any residual deformities remain, and so to confirm the natural history of pediatric trigger thumb. METHODS: We prospectively followed 87 thumbs in 67 patients with pediatric trigger thumb and these patients didn't receive any treatment such as passive stretching, splinting or surgery. The date of the first visit ranged from April 1994 to March 2005. The patients were evaluated every six-months prior to resolution and annually after resolution. The median duration of follow-up was 87.3 months (range, 60 to 156 months). RESULTS: Of the 87 trigger thumbs, 66 (75.9%) resolved spontaneously. The median time from the initial visit to resolution was 49.0 months (95% confidence interval, 41.1 to 56.9). There were no residual deformities that resolved beyond 48 months. Although complete resolution did not occur in the remaining 21 thumbs, the flexion deformities did improve in all 21 thumbs. There were no other differences between the two groups besides the average duration of follow-up. There was no difference in resolution based on gender. CONCLUSIONS: Pediatric trigger thumb can spontaneously resolve in > 75% of the cases after a follow-up period of at least 5 years. An operation may be delayed or avoided in the majority of cases. This may help both the families and the surgeons make decisions regarding the proper treatment of pediatric trigger thumb. PMID- 21629479 TI - Lordoplasty: an alternative technique for the treatment of osteoporotic compression fracture. AB - We report here on a new technique using polymethylmethacrylate to manage vertebral osteoporotic compression fractures in three patients. These patients presented with acute back pain that manifested itself after minor trauma. Osteoporotic compression fractures were diagnosed via plain X-ray and magnetic resonance imaging studies. The patients were treated with absolute bed rest and non-steroidal anti-inflammatory drugs. Despite of the conservative treatment, the patients experienced severe, recalcitrant and progressive pain. The vertebrae were collapsed over 50% or kyphotic deformity was seen on the radiologic materials. We performed a new technique called lordoplasty, which is derived from percutaneous vertebroplasty. The patients experienced a reduction in pain after the procedure. The wedge and kyphotic angles of the fractured vertebrae were significantly restored. PMID- 21629480 TI - Spontaneous rupture of the extensor pollicis longus tendon in a tailor. AB - A spontaneous rupture of the extensor pollicis longus (EPL) tendon is associated with rheumatoid arthritis, fractures of the wrist, systemic or local steroids and repetitive, and excessive abnormal motion of the wrist joint. The authors encountered a case of a spontaneous rupture of the EPL tendon. The patient had no predisposing factors including trauma or steroid injection. Although the patient had a positive rheumatoid factor, he did not demonstrate other clinical or radiological findings of rheumatoid arthritis. During surgery, the EPL tendon was found to be ruptured at the extensor retinaculum (third compartment). Reconstruction of the extensor tendon using the palmaris longus tendon was performed. At the 18-month follow-up, the patient showed satisfactory extension of the thumb and 40 degrees extension and flexion at the wrist. PMID- 21629481 TI - Ex vivo observation of human nucleus pulposus chondrocytes isolated from degenerated intervertebral discs. AB - STUDY DESIGN: We performed an ex vivo study to observe cell morphology and viability of human nucleus pulposus (NP) chondrocytes isolated from degenerated intervertebral discs (IVD). PURPOSE: To better understand the biological behavior of NP chondrocytes in monolayer cultures. OVERVIEW OF LITERATURE: Biological repair of IVDs by cell-based therapy has been shown to be feasible in clinical trials. As one of the most promising transplanting seeds, how the isolated NP chondrocytes behavior ex vivo has not been fully understood. METHODS: Human NP chondrocytes were harvested from 20 degenerated IVDs and cultured in monolayers. Histological and immunochemistry staining was used to detect cell morphology change. Cell viability was studied by analyzing cell cycle distribution and apoptotic rate in the primary and subculuted cells. RESULTS: The round or polygonal primary NP chondrocytes had an average adherence time of 7 days and took nearly 31 days to reach 95% confluence. The spindle-shaped P1 NP chondrocytes increased growth kinetics and took about 12 hours to adhere and 6.6 days to get 95% confluent. Immunochemistry staining of collagen II was positive in the cell cytoplasm. Nearly 90% of the confluent NP chondrocytes stayed in G1 phase while 16% underwent apoptosis. No significant difference of the collagen II expression, cell cycle distribution or the apoptosis indices were detected between the primary and subcultured NP chondrocytes. CONCLUSIONS: Human NP chondrocytes undergo significant morphological change in monolayer cultures. Cell cycle distribution pattern and apoptosis index of the cutured NP chondrocytes potentially influence their clinical transplantation or laboratory use. PMID- 21629482 TI - Adjacent Segment Degeneration after Single-Level PLIF: Comparison between Spondylolytic Spondylolisthesis, Degenerative Spondylolisthesis and Spinal Stenosis. AB - STUDY DESIGN: A retrospective study. PURPOSE: To comparatively investigated the rate of the adjacent segment degeneration and the clinical outcomes in patients with spondylolytic spondylolisthesis, spinal stenosis or degenerative spondylolisthesis. OVERVIEW OF LITERATURE: There have been few studies reported on the adjacent segment degeneration following posterior lumbar interbody fusion(PLIF). Many risk factors for the adjacent segment degeneration following PLIF have been proposed. The range of decompression has been presented as one of the risk factors, yet controversial. METHODS: This study enrolled sixty-three patients who had been treated with single-level PLIF and who were followed up for more than two years. The patients were divided into 3 groups based on the preoperative diagnosis. We analyzed the difference between the preoperative and postoperative intervertebral disc heights of the superior adjacent segments. The incidence rates of instability and the clinical outcomes were comparatively analyzed between each group. RESULTS: The average age of the patients was 55.8 years in the spondylolytic spondylolisthesis group, 65.9 years in the degenerative spondylolisthesis group and 60.4 years in the spinal stenosis group. The average follow-up period was 44 months, 43 months and 42 months, respectively. At the last follow-up, compared to the preoperative period, the intervertebral disc height decreased in all three groups. A statistically significant decrease (p < 0.01) was observed only in the spondylolytic spondylolisthesis group and no significant difference was observed between each group (p = 0.41). The incidence rate of instability and the clinical outcome were not significantly different between each group. CONCLUSIONS: Spondylolytic spondylolisthesis with total laminectomy and single-level PLIF showed no significant difference in the superior adjacent segment degeneration and instability, and the clinical outcome as compared to that of partial laminectomy with single-level PLIF for treating degenerative spondylolisthesis or spinal stenosis. PMID- 21629483 TI - The impact of fentanyl matrix on pain and function in spinal disorder-related chronic pain: an open label trial in Korea. AB - STUDY DESIGN: This is a multicenter, open-label prospective, non interventional study. PURPOSE: We wanted to evaluate the impact of fentanyl matrix on the pain and function of patients with spinal disorder-related chronic, non-malignant pain. OVERVIEW OF LITERATURE: Patients with severe non-malignant chronic low back pain may require opioid analgesics for effective pain management. METHODS: A total of 1,576 patients with severe pain (numeric rating scale = 7) were evaluated for their pain intensity at the initial visit and at weeks 4 and 8 (Visits 1, 2, and 3, respectively). Disturbances in sleep, daily living and social activities, the Oswestry Disability Index (ODI), the researchers' and patients' global assessment and the patients' treatment preference were also assessed. RESULTS: The pain intensity score significantly decreased from 8.1 at Visit 1 to 5.4 and 4.4 at Visits 2 and 3, respectively. Sleep disturbance also significantly decreased and the extent of disturbance of daily and social activities was also significantly improved. The ODI significantly decreased from 61.9% to 45.8% and 38.2% at Visits 1, 2, and 3, respectively. Adverse events were reported by 197 (12.5%) patients and severe adverse events were reported by 12 (0.76%) patients. Overall, 76.3% of the patients and 78.4% of the investigators rated the test drug as effective. CONCLUSIONS: The fentanyl matrix is believed to be effective for the treatment of pain, sleep disturbance and the impact upon daily and social activities, yet physicians should pay attention to the risks of abuse and the adverse events. PMID- 21629484 TI - Posterior decompression and fusion in patients with multilevel lumbar foraminal stenosis: a comparison of segmental decompression and wide decompression. AB - STUDY DESIGN: This is a prospective study. PURPOSE: We compared the outcomes of segmental decompression and wide decompression in patients who had multilevel lumbar foraminal stenosis with back pain. OVERVIEW OF LITERATURE: Wide decompression and fusion in patients with multilevel lumbar foraminal stenosis may increase the risk of perioperative complications. METHODS: From March 2005 to December 2007, this study prospectively examined 87 patients with multilevel lumbar foraminal stenosis and who were treated by segmental or wide decompression along with posterior fusion using pedicle screw fixation, and these patients could be followed-up for a minimum of 2 years. Of the 87 patients, 45 and 42 patients were assigned to the segmental decompression group (group 1) and the wide decompression group (group 2), respectively. We compared the clinical and radiological outcomes of the patients in these two groups. RESULTS: There were no significant differences between groups 1 and 2 in terms of the levels of postoperative pain based on the visual analogue scale, the Oswestry Disability Score, the clinical results based on the Kirkaldy-Willis Criteria, the complication rate or the posterior fusion rate. On the other hand, the mean operating times in groups 1 and 2 were 153 +/- 32 minutes and 187 +/- 36 minutes, respectively (p < 0.05). The amount of blood loss during surgery and on the first postoperative day was 840 +/- 236 ml and 1,040 +/- 301 ml in groups 1 and 2, respectively (p < 0.05). CONCLUSIONS: These results suggest that segmental decompression offers promising and reproducible clinical and radiological results for patients suffering from multilevel lumbar foraminal stenosis. PMID- 21629485 TI - Are we missing osteoporosis-related vertebral fractures in men? AB - STUDY DESIGN: Retrospective study. PURPOSE: To assess the prevalence of osteoporosis related spinal fractures among Saudi Arabian males. OVERVIEW OF LITERATURE: Vertebral fractures are the most common complication of osteoporosis and is the first sign in both sexes and only 25 to 30% of radiographically observed vertebral deformities are recognized. METHODS: We analyzed the chest radiographs of consecutive Saudi Arabian men >= 50 years and who visited the emergency room of King Fahd University Hospital, Al Khobar, Saudi Arabia for a period of 12 months between November 1, 2007 and October 31, 2008. The site and type of fractures were classified as per the semi-quantitative technique. The other data retrieved from the medical records of patients included medications and clinical investigations for osteoporosis. RESULTS: Nine hundred seventy chest radiographs were performed during the study period and 876 radiographs could be analyzed. One hundred fifteen patients (13.1%) had 157 fractures. The mean age was 67.85 +/- 10.1 years. There was more than one fracture in 21 patients (18.2%). The majority of fractures (n = 102, 64.9%) were observed in thoracic spine. Seventy-one (45.2%) fractures were classified as mild, 54 (34.4%) were moderate and 32 (20.4%) were severe. For 26 (22.6%) patients, the report of the radiologist highlighted the fracture. CONCLUSIONS: Saudi Arabian males with osteoporosis continue to be missed despite the high prevalence osteoporosis leading to vertebral fractures. We believe it is important for physicians to identify vertebral fractures early and treat then appropriately before an extremity fracture occurs with high mortality. PMID- 21629486 TI - The clinical and radiological outcomes of minimally invasive transforaminal lumbar interbody single level fusion. AB - STUDY DESIGN: This is a retrospective study that was done according to clinical and radiological evaluation. PURPOSE: We analyzed the clinical and radiological outcomes of minimally invasive transforaminal lumbar interbody single level fusion. OVERVIEW OF LITERATURE: Minimally invasive transforaminal lumbar interbody fusion is effective surgical method for treating degenerative lumbar disease. METHODS: The study was conducted on 56 patients who were available for longer than 2 years (range, 24 to 45 months) follow-up after undergoing minimally invasive transforminal lumbar interbody single level fusion. Clinical evaluation was performed by the analysis of the visual analogue scale (VAS) score and the Oswestry Disability Index (ODI) and the Kirkaldy-Willis score. For the radiological evaluation, the disc space height, the segmental lumbar lordotic angle and the whole lumbar lordotic angle were analyzed. At the final follow-up after operation, the fusion rate was analyzed according to Bridwell's anterior fusion grade. RESULTS: For the evaluation of clinical outcomes, the VAS score was reduced from an average of 6.7 prior to surgery to an average of 1.8 at the final follow-up. The ODI was decreased from an average of 36.5 prior to surgery to an average of 12.8 at the final follow-up. In regard to the clinical outcomes evaluated by the Kirkaldy-Willis score, better than good results were obtained in 52 cases (92.9%). For the radiological evaluation, the disc space height (p = 0.002), and the whole lumbar lordotic angle (p = 0.001) were increased at the final follow-up. At the final follow-up, regarding the interbody fusion, radiological union was obtained in 54 cases (95.4%). CONCLUSIONS: We think that if surgeons become familiar with the surgical techniques, this is a useful method for minimally invasive spinal surgery. PMID- 21629487 TI - Post-operative Changes of Cerebral Glucose Metabolism in Patients with Lumbar Spinal Stenosis with Pre-operative Anxiety: Statistical Parametric Mapping Analysis of F-18 FDG Brain PET. AB - STUDY DESIGN: A prospective study. PURPOSE: To assess postoperative changes in cerebral glucose metabolism in anxiety patients with lumbar spinal stenosis (SS). OVERVIEW OF LITERATURE: Although an association between preoperative anxiety and abnormal cerebral glucose metabolism may exist, only a limited number of studies using F-18 fluorodeoxyglucose positron emission tomography (FDG PET) have evaluated preoperative to postoperative changes in cerebral glucose metabolism in SS patients in detail. METHODS: The present study was designed to assess preoperative to postoperative changes in cerebral glucose metabolism in anxiety patients with SS. F-18 FDG PET with statistical parametric mapping analyses was used to compare preoperative and postoperative regional brain glucose metabolism in 18 SS patients. RESULTS: F-18 FDG PET scans showed postoperative activation of several brain clusters in gray matter. These included left parahippocampus, left cerebellar tonsil, left inferior semi-lunar lobule, and right cerebellar tonsil. Areas that were deactivated postoperatively were the right insula, left fusiform gyrus, left orbitofrontal cortex, left inferior frontal gyrus, left middle frontal gyrus, left precuneus, and left inferior frontal gyrus. CONCLUSIONS: SS patients with preoperative anxiety showed altered cerebral glucose metabolism at postoperative follow-up. PMID- 21629488 TI - A Broken Drill-bit Fragment Causing Severe Radiating Pain after Cervical Total Disc Replacement: A Case Report. AB - This is a case report of a 38-year-old man with severe radiating pain on upper extremity after cervical total disc replacement (TDR). We faced an unusual complication that has not been reported yet. He underwent cervical TDR for left central disc protrusion on C5-6. After the surgery, preoperative symptom disappeared. However, at postoperative 1 year, he complained severe right-sided radiating pain that had a sudden onset. On postoperative X-ray, a metal fragment which seemed like a broken drill bit was shown within the spinal canal. To remove that, right-sided anterior microforaminotomy on C5-6 was performed and the metal fragment was removed successfully. After that, anterior fusion was done because the motion of the artificial disc was minimal and the removed structure seemed to attenuate stability during cervical motion. The operation resulted in prompt symptomatic relief. During cervical TDR, particular attention should be paid to the procedures that require using drill-bits. PMID- 21629489 TI - Post-traumatic Lumbar Epidural Hematoma with Neurology: Report of 1 Case. AB - The purpose of this study was to report a case with post-traumatic spinal epidural hematomas with abnormal neurologic findings, which is uncommon. A 40 year-old man presented at our clinic after a blunt trauma caused by a traffic accident in which he was a pedestrian. After admission, abnormal neurologic symptoms developed including loss of sensation and motor function in his left lower extremity. Magnetic resonance imaging demonstrated a spinal epidural hematoma with 40% canal stenosis at the L5-S1 level. Decompression including hematoma evacuation was done. Symptoms started to be reduced 18 days after operation. He was treated conservatively with medications and all symptoms resolved completely during admission and there were no further neurologic sequelae. Post-traumatic lumbar spinal epidural hematoma with abnormal neurologic findings is an uncommon condition that may present belatedly after trauma with significant neurologic compromise. PMID- 21629490 TI - Pediatric lumbar epidural abscess combined with cauda equina syndrome: case report. AB - Pyogenic epidural abscess is a very rare disease. Once it occurs, it promptly progresses and can cause neurologic paralysis. Mean age of onset has been reported to be 57 years. Here we report making a diagnosis of pyogenic lumbar epidural abscess accompanying cauda equina syndrome in a 10-year-old girl. We treated this case successfully with surgical drainage and antibiotics. We report our case with a review of the literature. PMID- 21629491 TI - Reviews in obstetrics & gynecology goes global: building ties with china. PMID- 21629492 TI - Tubal-cervical twin pregnancy. PMID- 21629493 TI - Management of first trimester pregnancy loss can be safely moved into the office. AB - MANAGEMENT OF FIRST TRIMESTER PREGNANCY LOSS HAS CONVENTIONALLY INVOLVED TWO OPTIONS: expectant management or dilation and curettage in the operating room. New options in the outpatient setting are providing women with alternatives that can be less expensive and performed in more private settings. This review discusses the available approaches to expectant, medical, and surgical management of first trimester loss and the comparative efficacy of each method. PMID- 21629495 TI - Maternal health considerations during disaster relief. AB - When disasters strike resource-poor nations, women are often the most affected. They represent the majority of the poor, the most malnourished, and the least educated, and they account for more than 75% of displaced persons. The predisaster familial duties of women are magnified and expanded, and they have significantly less support and fewer resources than they had before the incident. Moreover, after the disaster, they bear the responsibility of caring for their children, the elderly, the injured, and the sick. Besides the effects of the disaster, women become more vulnerable to reproductive and sexual health problems and are at increased risk for physical and sexual violence. Women become both victims and the primary caretakers. Health practitioners are often not aware of these issues when providing emergency care. Developing a disaster relief team with experts in maternal health is necessary to improve women's health outcome. PMID- 21629494 TI - Ovarian cancer screening and early detection in the general population. AB - Worldwide, the estimated annual incidence of ovarian cancer is 204,000, with 125,000 deaths. In developed countries, ovarian cancer remains the most lethal of all gynecologic malignancies. One of the reasons for the high fatality rate is that more than 70% of women with ovarian cancer are diagnosed with advanced disease. There is a close correlation between stage at presentation and survival; therefore, early detection of ovarian cancer represents the best hope for mortality reduction and long-term disease control. There is preliminary evidence that screening can improve survival, but the impact of screening on mortality from ovarian cancer is still unclear. The proteomic approach has yielded encouraging preliminary findings, but these findings are not mature enough for clinical use. At this time, clear recommendations cannot be made on the basis of the available data. PMID- 21629496 TI - The role of mechanical bowel preparation in gynecologic laparoscopy. AB - Various combinations of dietary restriction, antibiotic regimens, and mechanical preparations have become routine in preoperative surgical planning for elective colon surgery. This practice has also become commonplace in the field of gynecology, either for planned bowel surgery or in complex cases that are believed to be high risk for inadvertent bowel injury. As the trend in gynecologic surgery shifts toward more minimally invasive approaches, the complexity of cases being performed by laparoscopy and robotics continues to increase. In addition, laparoscopic surgical techniques have a different set of inherent risks and challenges as compared with open pelvic operations. This review summarizes the available data surrounding the use of mechanical bowel preparations, specifically with regard to gynecologic laparoscopy. PMID- 21629497 TI - Home deliveries and risk. PMID- 21629498 TI - Induction and cost. PMID- 21629499 TI - Apgar scores and cerebral palsy. PMID- 21629500 TI - Estradiol vaginal tablets. PMID- 21629501 TI - Snippets. PMID- 21629502 TI - A quarter of a century with AIDS. AB - In Northwestern Spain (NWS), the annual incidence of AIDS diagnoses increased from 1984 (when the first case was diagnosed) until 1996. However, since 1996, this incidence has reduced considerably, including a notable 40% reduction between 1997 and 1998. The Galician Register of AIDS supplies information on the evolution of AIDS pathology in NWS. This report compiles data on patients who were diagnosed with AIDS in NWS between 1984 and 2008. From 1981, when the first case of AIDS was described, until December 31, 2008, a total of 3,766 AIDS cases were registered in NWS. Of these, 2,085 cases (55.4%) resulted in death. Examining data from individual provinces revealed that the highest number of cases was in A Coruna (1,548 cases) followed by Pontevedra (1,485 cases).For almost half of the new cases of AIDS diagnosed between 2003 and 2008 (44%), less than six months passed between the diagnosis of infection and manifestations of the disease. Thus, the number of patients that do not receive early diagnosis of HIV infection has remained high.With regard to the transmission mechanism, 64% of the cases occurring during these years resulted from needle-sharing among injected drug users (IDUs). Unprotected heterosexual and homosexual practices were responsible for 20% and 17% of the cases, respectively. PMID- 21629503 TI - HIV/AIDS Clinical Manifestations and their Implication for Patient Clinical Staging in Resource Limited Settings in Tanzania. AB - BACKGROUND: Tanzania HIV/AIDS management follows WHO clinical staging which requires CD4 counts as complement. Lacking CD4 counts facilities in rural health facilities remains a challenge. Simplified and sensitive clinical staging based on local clinical patterns is useful to ensure effective care without CD4 counts. OBJECTIVES: To assess whether local HIV clinical manifestations can be used to guide HIV management in settings with limited access to CD4 counts in Tanzania. METHODS: A Cross-sectional study conducted at Tumbi and Chalinze health facilities documented clinical manifestations and CD4 counts in 360 HIV/AIDS patients. Simplified management groups comprised of severe and moderate disease were formed based on clinical manifestations and CD4 counts results. Symptoms with high frequency were used to predict severe disease. RESULTS: A Weight loss (48.3%) and chronic cough (40.8 %) were the most reported manifestations in the study population. More than 50% of patients presented with CD4<=200. Most symptoms were found to be highly sensitive (71% to 93%) in predicting severe immunosuppression using CD4<200 cut-off point as a 'Gold standard'. Chronic diarrhoea presented in 10.6%, and predicted well severe immunosuppression either alone (OR 1.95, 95%CI, 0.95-4.22) or in combination (OR 4.21, 95%CI 0.92-19.33) with other symptoms. Basing strictly on WHO clinical staging 30.8% of patients were detected to be severely immunosuppressed (Stage 4). While using our proposed management categories of severe and moderate immunosuppression 70% of patients were put into the severe immunosuppression group, consistent with CD4 cut-off count of<=350. CONCLUSIONS: HIV/AIDS clinics managing large cohorts should develop validated site specific guidelines based on local experiences. Simplified guidelines are useful for resource constrained settings without CD4 counting facilities. PMID- 21629504 TI - Multiple ART Programs Create a Dilemma for Providers to Monitor ARV Adherence in Uganda. AB - BACKGROUND: Increased availability and accessibility of antiretroviral therapy (ART) has improved the length and quality of life amongst people living with HIV/AIDS. This has changed the landscape for care from episodic to long-term care that requires more monitoring of adherence. This has led to increased demand on human resources, a major problem for most ART programs. This paper presents experiences and perspectives of providers in ART facilities, exploring the organizational factors affecting their capacity to monitor adherence to ARVs. METHODS: From an earlier survey to test adherence indicators and rank facilities as good, medium or poor adherence performances, six facilities were randomly selected, two from each rank. Observations on facility set-up, provider-patient interactions and key informant interviews were carried out. The strengths, weaknesses, opportunities and threats identified by health workers as facilitators or barriers to their capacity to monitor adherence to ARVs were explored during group discussions. RESULTS: Findings show that the performance levels of the facilities were characterized by four different organizational ART programs operating in Uganda, with apparent lack of integration and coordination at the facilities. Of the six facilities studied, the two high adherence performing facilities were Non-Governmental Organization (NGO) programs, while facilities with dual organizational programs (Governmental/NGO) performed poorly. Working conditions, record keeping and the duality of programs underscored the providers' capacity to monitor adherence. Overall 70% of the observed provider patient interactions were conducted in environments that ensured privacy of the patient. The mean performance for record keeping was 79% and 50% in the high and low performing facilities respectively. Providers often found it difficult to monitor adherence due to the conflicting demands from the different organizational ART programs. CONCLUSION: Organizational duality at facilities is a major factor in poor adherence monitoring. The different ART programs in Uganda need to be coordinated and integrated into a single well resourced program to improve ART services and adherence monitoring. The focus on long-term care of patients on ART requires that the limitations to providers' capacity for monitoring adherence become central during the planning and implementation of ART programs. PMID- 21629505 TI - Learning from Interviews with HIV-Discordant Couples (Male Positive, Female Negative): The Challenges and Successes. AB - This article examines the challenges and successes of recruiting participants and maintaining momentum in a small qualitative study on the experiences of HIV discordant couples (where the male is HIV-positive and the female is HIV negative) undergoing fertility assessment and/or treatment in Ontario, Canada, to reduce the risk of HIV transmission to the woman and fetus. The purpose of this article is to identify barriers and successes encountered in our study, consider how these are addressed in the literature, and highlight specific factors that need to be considered when studying a unique population similar to ours. PMID- 21629506 TI - Drug development to protozoan diseases. AB - The diseases caused by protozoan parasite are responsible for considerable mortality and morbidity, affecting more than 500 million of people in the world. The epidemiological control of protozoan is unsatisfactory due to difficulties of vector and reservoir control; while the progress in the development of vaccine tends to be slow and arduous. Currently, the chemotherapy remains essential component of both clinical management and disease control programmer in endemic areas. The drugs in use as anti-protozoan agents were discovered over 50 years and a number of factors limit their utility such as: high cost, poor compliance, drug resistance, low efficacy and poor safety. In the recent years, the searches about the development of new drugs against protozoa parasite have been increased. This special issue of The Open Medicinal Chemistry Journal will present some of developments in this field with the aim to shown the significant advances in the discovery of new anti-protozoan drugs. PMID- 21629507 TI - Current developments in the therapy of protozoan infections. AB - Protozoan parasites cause serious human and zoonotic infections, including life threatening diseases such as malaria, African and American trypanosomiasis, and leishmaniasis. These diseases are no more common in the developed world, but together they still threaten about 40% of the world population (WHO estimates). Mortality and morbidity are high in developing countries, and the lack of vaccines makes chemotherapy the only suitable option. However, available antiparasitic drugs are hampered by more or less marked toxic side effects and by the emergence of drug resistance. As the main prevalence of parasitic diseases occurs in the poorest areas of the world, the interest of the pharmaceutical companies in the development of new drugs has been traditionally scarce. The establishment of public-private partnerships focused on tropical diseases is changing this situation, allowing the exploitation of the technological advances that took place during the past decade related to genomics, proteomics, and in silico drug discovery approaches. These techniques allowed the identification of new molecular targets that in some cases are shared by different parasites. In this review we outline the recent developments in the fields of protease and topoisomerase inhibitors, antimicrobial and cell-penetrating peptides, and RNA interference. We also report on the rapidly developing field of new vectors (micro and nano particles, mesoporous materials) that in some cases can cross host or parasite natural barriers and, by selectively delivering new or already in use drugs to the target site, minimize dosage and side effects. PMID- 21629508 TI - Screening of Potential anti-Trypanosoma cruzi Candidates: In Vitro and In Vivo Studies. AB - Chagas disease (CD), caused by the intracellular protozoan Trypanosoma cruzi, is a parasitic illness endemic in Latin America. In the centennial after CD discovery by Carlos Chagas (1909), although it still represents an important public health problem in these affected areas, the existing chemotherapy, based on benznidazole and nifurtimox (both introduced more than four decades ago), is far from being considered ideal due to substantial toxicity, variable effect on different parasite stocks and well-known poor activity on the chronic phase. CD is considered one of the major "neglected" diseases of the world, as commercial incentives are very limited to guarantee investments for developing and discovering novel drugs. In this context, our group has been pursuing, over the last years, the efficacy, selectivity, toxicity, cellular targets and mechanisms of action of new potential anti-T. cruzi candidates screened from an in-house compound library of different research groups in the area of medicinal chemistry. A brief review regarding these studies will be discussed, mainly related to the effect on T. cruzi of (i) diamidines and related compounds, (ii) natural naphthoquinone derivatives, and (iii) megazol derivatives. PMID- 21629509 TI - Structures, targets and recent approaches in anti-leishmanial drug discovery and development. AB - Recent years have seen a significant improvement in available treatment options for leishmaniasis. Two new drugs, miltefosine and paromomycin, have been registered for the treatment of visceral leishmaniasis (VL) in India since 2002. Combination therapy is now explored in clinical trials as a new treatment approach for VL to reduce the length of treatment and potentially prevent selection of resistant parasites. However there is still a need for new drugs due to safety, resistance, stability and cost issues with existing therapies. The search for topical treatments for cutaneous leishmaniasis (CL) is ongoing. This review gives a brief overview of recent developments and approaches in anti leishmanial drug discovery and development. PMID- 21629510 TI - HIV Protease Inhibitors: Effect on the Opportunistic Protozoan Parasites. AB - The impact of highly active antiretroviral therapy (HAART) in the natural history of AIDS disease has been allowed to prolong the survival of people with HIV infection, particularly whose with increased HIV viral load. Additionally, the antiretroviral therapy could exert a certain degree of protection against parasitic diseases. A number of studies have been evidenced a decrease in the incidence of opportunistic parasitic infections in the era of HAART. Although these changes have been attributed to the restoration of cell-mediated immunity, induced by either non-nucleoside reverse transcriptase inhibitors or HIV protease inhibitors, in combination with at least two nucleoside reverse transcriptase inhibitors included in HAART, there are evidence that the control of these parasitic infections in HIV-positive persons under HAART, is also induced by the inhibition of the proteases of the parasites. This review focuses on the principal available data related with therapeutic HIV-protease inhibitors and their in vitro and in vivo effects on the opportunistic protozoan parasites. PMID- 21629511 TI - The emotional and neurological consequences of abuse. PMID- 21629512 TI - Maltreatment in childhood and adolescence: results from a survey of a representative sample of the German population. AB - BACKGROUND: There are no up-to-date, representative studies on the frequency of maltreatment (abuse or neglect) among children and adolescents in Germany. METHODS: In a cross-sectional study, standardized questionnaires were administered to persons aged 14 and older in a representative sample of the German population. Statistics on maltreatment in childhood and adolescence were collected with the German version of the Childhood Trauma Questionnaire. Regression analysis was used to detect potential associations of maltreatment with demographic variables including sex, age, place of birth, and social class. RESULTS: 2504 of the 4455 persons contacted (56%) completed the study. Severe emotional abuse in childhood and/or adolescence was reported by 1.6% of persons in the overall sample, severe physical abuse by 2.8%, and severe sexual abuse by 1.9%. Severe emotional neglect was reported by 6.6% and severe physical neglect by 10.8%. Female sex was a predictor for severe sexual abuse, while belonging to a low or middle social stratum was a predictor for severe physical abuse and neglect. Being older at the time of the survey was a predictor for severe physical neglect. All types of maltreatment were significantly correlated with each other (p<0.001). CONCLUSION: The frequencies of various types of abuse and neglect of children and adolescents that were retrospectively determined in this up-to-date study by questionnaire of a representative sample of the German population, and the correlations between them, correspond to those found in a German population-based study in 1995 and in recent American studies. PMID- 21629513 TI - Recurrent abdominal pain in childhood. AB - BACKGROUND: Chronic, recurrent abdominal pain is common among children and adolescents. It interferes with everyday life, causes absence from school, and leads to frequent medical consultations, often involving burdensome diagnostic testing and protracted attempts at treatment. METHOD: Selective review of the literature. RESULTS: Organic causes should be ruled out with a thorough medical history and physical examination and a small number of laboratory tests. The pediatric Rome III criteria include valid diagnostic criteria for functional abdominal pain in childhood. The available data imply that this condition is best treated with cognitive behavioral therapy, rather than with medications or dietary measures. CONCLUSION: A systematic approach to chronic recurrent abdominal pain in children and adolescents is key to ruling out organic diseases while avoiding unnecessary tests and treatments. PMID- 21629514 TI - Neuroendocrine neoplasms of the gastrointestinal tract. AB - BACKGROUND: Gastroenteropancreatic neuroendocrine neoplasms (GEP-NENs) are complex tumors whose incidence is rising and whose treatment requires precise classification and risk stratification. METHOD: Selective review of the relevant literature, including recently published guidelines. RESULTS: GEP-NENs are initially classified by their degree of histological differentiation and their graded cell proliferation (Ki-67 index). In addition, there are GEP-NEN specific TNM staging protocols. The laboratory assessment includes the measurement of general tumor markers (synaptophysin, chromogranin A) as well as specific ones (hormones). The most important imaging technique for diagnosis is octreotide scintigraphy. The surgical treatment of GEP-NEN is based on oncological resection criteria whose aim is to achieve locally radical resection while preserving as much organ function as possible. Metastases, too, may be amenable to resection. The treatment options for unresectable metastases include radiofrequency ablation and chemoembolization, both of which are palliative methods of reducing tumor volume and hormone production. Other chemotherapeutic and nuclear-medical treatments can be applied depending on the extent of metastatic spread, the proliferation index, and the degree of hormone production by the tumor. CONCLUSION: The accurate diagnosis and appropriate treatment of GEP-NET currently gives most patients with this tumor a good prognosis, as long as it is discovered early. Early GEP-NETs have a favorable prognosis. Further advances in the diagnosis and treatment of this disease may result from structural changes in patient care, including the establishment of NET centers. PMID- 21629515 TI - Barrett's adenocarcinoma of the esophagus: better outcomes through new methods of diagnosis and treatment. AB - BACKGROUND: Esophageal adenocarcinoma has attracted more attention among gastroenterologists recently because of its rapidly rising incidence in Western countries. Many new epidemiological findings have been published, and there have been numerous technical advances in diagnostic procedures and in multimodal treatment based on the staging of the disease. METHODS: In this paper, we selectively review the literature on esophageal adenocarcinoma, also considering the evidence-based recommendations contained in the guidelines of the German Society for Digestive and Metabolic Diseases (Deutsche Gesellschaft fur Verdauungs- und Stoffwechselkrankheiten, DGVS) as well as the latest data from our own research team. RESULTS AND CONCLUSION: here have been major recent advances in the diagnosis and treatment of esophageal adenocarcinoma. New refinements in endoscopic techniques now make endoscopic treatment possible for early esophageal carcinoma. New surgical techniques and new strategies of neoadjuvant chemotherapy have lowered the morbidity and improved the outcome of patients with locally advanced disease. Molecular therapies, too, have shown promising initial results. PMID- 21629516 TI - Correspondence (letter to the editor): Non-albicans Candida species. PMID- 21629517 TI - Correspondence (reply): In Reply. PMID- 21629518 TI - Correspondence (letter to the editor): Association studies often lack plausibility: back to the future of mechanisms. PMID- 21629519 TI - Correspondence (letter to the editor): Benefits outweigh risks. PMID- 21629520 TI - Correspondence (reply): In Reply. PMID- 21629521 TI - Pharmacological approaches for management of child and adolescent obesity. AB - Overweight and obesity among children and adolescents continue to be a global public health epidemic. Current national data on childhood and adolescent obesity show alarming statistics of overweight and obese children and adolescents. This epidemic runs across various continents and affects various ethnic populations. The current weight management practices involve dietary modification, behavioral change therapies involving exercise, pharmacological therapy and surgical intervention. The purpose of this current review is to focus on pharmacological therapy i.e. role of sibutramine and orlistat in childhood and adolescent weight management. An open search of PUBMED database was made with search key words such as orlistat or sibutramine or pharmacological approaches and child obesity and adolescent obesity. This yielded a total of 20 articles. All of these articles have been summarized in the current review. Sibutramine functions by promoting satiety and increases energy expenditure by inhibiting reuptake of noradrenaline and serotonin. Most of the studies, reviews and trials conducted using sibutramine among adolescents and children show limited short-term efficacy. The long-term effects of sibutramine use are not yet studied due to the severity of its side-effects profile. Orlistat was approved by Food and Drug Administration for adolescent weight reduction. Despite its approval, it has a limited role in adolescent and pediatric obesity reduction due to the purported malabsorption of fat soluble vitamins and its side-effect profile. Obesity and adolescent weight management in present times mainly deals with dietary modification with superadded behavioral therapies promoting exercise. There is insufficient evidence as of now if these alone would cause adequate weight reduction and weight maintenance. Pharmacotherapy i.e use of drugs like sibutramine and orlistat has a limited role in the current fight against childhood and adolescent obesity. Extreme side-effects, close monitoring and lack of long-term studies involving these drugs, suggest questionable efficacy in current times. Future research studies involving pharmacological agents need to not only have immense scientific rigor during preliminary analyses but should also translate their efficacy in practical and clinical settings. KEYWORDS: Obesity; Overweight; Adolescents; Pharmacological; sibutramine; Orlistat. PMID- 21629522 TI - Implementation of Screening for Toxoplasma gondii Infection in Pregnancy. AB - BACKGROUND: Since 1998, Italian law requires serological screening for toxoplasmosis by the thirteenth week of pregnancy, and seronegative women should undergo further checks every 30 - 40 days until delivery (a total of 5 - 7 screenings). This is an improvement of the previous law which foresaw three free tests (one by the end of the third month, one in the fifth, and one in the ninth month of pregnancy). The aim of this study was to assess the implementation, in an urban area of Northern Italy, of the 1998 law about 10 years after its entry into force. METHODS: Of the 4,694 women who initiated and completed a pregnancy in the period 2006 - 2008, we recorded the trimester of pregnancy in which they underwent their first screening, the total and average number of screenings during pregnancy, and the trimester distribution of the screenings. RESULTS: A total of 84.1% of the women underwent their first screening during the first trimester. The negative cases underwent an average of 3.7 screenings during pregnancy, with 34.9% undergoing five or more. Sixty percent of the women underwent at least one screening per trimester. CONCLUSIONS: Our data indicates active screening during the first trimester, but fewer screenings than required by law during pregnancy as a whole. Therefore further efforts are needed to improve screening implementation. KEYWORDS: Anti-Toxoplasma antibodies; Congenital infection; Pregnancy trimester; Toxoplasma screening; Seroconversion; Seronegative women; Seropositive women; Screening protocol. PMID- 21629523 TI - Latent toxoplasmosis in patients with different malignancy: a hospital based study. AB - BACKGROUND: Seroprevalence of toxoplasmosis in different populations may vary according to different environments, social customs and habits. This study was designed to measure the seroprevalence of toxoplasmosis among patients with different malignancies and to ascertain the association between common risk factors and disease transmission. METHODS: This Cross-sectional study was from January to April of 2009. Four Oncology wards in Hospital Universiti Kebangsaan Malaysia (HUKM) were selected as the site for undertaking the present study. The survey involved 129 patients with different malignancies. Information was gathered by using study subject information sheet and a standardized structured questionnaire. Toxoplasma was screened by a standerd ELISA commercial kit in accordance with the manufacturers instructions and performed at the Department of Microbiology, HUKM Kuala Lumpur. A result of > 51 IU/ml of anti-Toxoplasma (IgG) antibody was regarded as positive, indicating latent or pre-existing Toxoplasma infection. A result of > 51 IU/ml of anti-Toxoplasma (IgM) antibody was regarded as positive, indicating recently acquired Toxoplasma infection. RESULTS: Total number of seropositive patients was 54 (67.6%), the mean age was 51 years (range15 - 88 years). Toxoplasma IgG positivity was highest among Malaysian (32%). Male to female ratio was almost equal. There was a statistically significant difference in seropositivity between patients living in rural areas compared to those living in urban areas, positive history of consumption of undercooked meat and/or blood transfusion (p < 0.05). CONCLUSIONS: These findings give some support to Toxoplasma screening program and health education, including promotion of a healthy lifestyle exclusively in seronegative patients in order to prevent seroconversion and the incidence of clinically evident opportunistic infection. KEYWORDS: Toxoplasma gondii; Risk factors; Immunocompromised; seroconversion; Seroprevalence. PMID- 21629524 TI - Sensitivity, specificity, negative and positive predictive values of adenosine deaminase in patients of tubercular and non-tubercular serosal effusion in India. AB - BACKGROUND: In India, tuberculosis is an endemic disease. Delay in diagnosis results in poor prognosis and fast spread of the disease. The objective of the present study is to look for an effective and acceptable diagnostic test, which may be helpful to initiate early treatment to improve prognosis and reduce spread. METHODS: Three hundred and thirty patients with pleural, ascitic, meningeal and synovial effusion were selected and divided depending upon the etiology and the involvement of serosal membranes. Serosal aspirated fluid was subjected to biochemical tests and adenosine deaminase estimation. Cutoff taken is above 40 for pleural, peritoneal or synovial fluid and above 10 for CSF. RESULTS: In cases of pulmonary and extra-pulmonary disease, sensitivity was 92.80% and 94.29%; specificity 90.00% and 92.16%; positive predictive value 92.86% and 89.00%; and negative predictive value 90.00% and 95.92% respectively. CONCLUSIONS: Adenosine deaminase estimation is not only a fairly sensitive and specific test (more than 90%), helpful in differentiating tubercular from non tubercular etiology both in pulmonary and extra-pulmonary disease, but is also simple, inexpensive and rapid. For this reason this test may help in early diagnosis, improve the prognosis and reduce spread of disease and sequlae. KEYWORDS: Adenosine deaminase; Serosal effusion; Tubercular; Non-tubercular; Pulmonary; Extra-pulmonary. PMID- 21629525 TI - Evaluation of Transcutaneous Electrical Nerve Stimulation as a Treatment of Neck Pain due to Musculoskeletal Disorders. AB - BACKGROUND: This study was designed to evaluate transcutaneous electrical nerve stimulation (TENS) as a treatment for neck pain due to musculoskeletal disorders within the context of a physiotherapy treatment. METHODS: Thirty subjects with neck pain were randomly allocated to two groups, treated with either TENS (n = 15) or placebo (n = 15). Each subject received one session for one hour. All subjects were evaluated before, during treatment, after switch off and again a week after by using Myometer machine. All subjects completed the follow-up assessment. Subjects referred for out-subjects' physiotherapy department, fulfilling the inclusion and exclusion criteria, took part in the study. RESULTS: The assessments were compared and used to measure outcome treatment. Improvement in their condition was measured in terms of a reduction in the individual's level of pain during the week after the end of the first session. At the end of the first session, the study showed that 11 subjects (73%) in the treatment and 7 subjects (43%) in the control groups had gained marked improvement. These results are statistically highly significant, (P = 0.01) at the end of the follow-up assessment. CONCLUSIONS: A conclusion could be drawn that a single intense TENS treatment is an effective treatment for neck pain due to musculoskeletal disorders. On the other hand, TENS showed an effective pain relief with subjects who have a mild neck pain rather than those with severe symptoms. KEYWORDS: Musculoskeletal disorders; Transcutaneous electrical nerve stimulation; Neck pain. PMID- 21629526 TI - Rectal metastasis of prostate cancer: about a case. AB - Prostate adenocarcinomas present a high risk of metastasis. We report a case of an atypical prostate cancer metastasis. A male patient presented a prostatic adenocarcinoma treated by surgery. A biological recurrence was discovered during the follow-up by an increased rate of Prostate Specific Antigen (PSA) and was treated by hormonotherapy. Several months later, there was a re-increase of the PSA rate. The CT scan showed a radiation proctitis aspect. An intermittent hormonotherapy was decided. Six months later, he presented abdominal pain. Examinations were performed and showed a rectal carcinosarcoma with prostate origins. A surgical management was realised. The outcomes were an early recurrence. A symptomatic treatment was decided. There are not any rectal localisations reported in the literature. Only loco-regional invasions of the rectum are described and no histological modification of metastasis compared to the primitive tumor has been reported. So, we report a metastasis of a prostate adenocarcinoma which transformed into a carcinosarcoma. KEYWORDS: Adenocarcinoma; Carcinosarcoma; Metastasis; Prostate; Rectal neoplasm. PMID- 21629527 TI - A case of intrathoracic stomach and spleen after aortic repair: an unusual complication. AB - A patient is described who presented with an intrathoracic stomach and spleen two weeks after aortic repair for an aortoduodenal fistula. At an urgent laparotomy the stomach was repositioned and the spleen removed. The patient recovered fully. The possible mechanism of this severe complication is discussed and relevant literature reviewed. KEYWORDS: Aortoduodenal fistula; Complication; Intrathoracic stomach; Intrathoracic spleen. PMID- 21629528 TI - Myositis ossificans circumscripta without history of trauma. AB - Myositis ossificans circumscripta is a form of heterotopic ossification that is benign in nature associated to a trauma, but may appear clinically and radiologically as a malignant neoplasm. We describe a rare case of calcifying of myositis ossificans not associated to trauma in a 35-year-old woman with a mass in her upper third and external of right thigh. We discuss some of the difficulties of diagnosis and histological evolution of the lesion. KEYWORDS: Myositis ossificans; Thigh; Differential diagnosis; Nontraumatic. PMID- 21629529 TI - Intensive care of a weils disease with multiorgan failure. AB - Leptospirosis is a commonly encountered type of zoonosis, especially in tropical regions. There is insufficient data regarding its frequency in non-tropical regions such as Turkey. Although leptospirosis presents with a mild icteric form in nearly 90% of cases, it can lead to Weils disease characterized by fever as well as fulminant hepatorenal and respiratory failure, in approximately 5 - 10% of cases. In this case report, we present a patient with Weil's disease, complicated with multiorgan failure. KEYWORDS: Weils disease; Leptospirosis; Multiorgan failure. PMID- 21629530 TI - Fundal variant adenomyomatosis of the gallbladder: report of three cases and review of the literature. AB - Adenomyomatosis of the gallbladder (adenomyomas or adenomyomatous hyperplasia) is relatively uncommon tumor or tumor-like lesions that are found in 2%5% of all cholecystectomies. They may involve in the fundal localization as a second frequency and have a predilection for the middle and elderly-aged women. Adenomyomas are histologically characterized by epithelial proliferation and muscular hyperplasia of the gallbladder. The chronic irritation is mostly aetiological factor and rarely has malignant potential. In this paper, the authors present the clinicopathological features of three cases with adenomyomatosis of the gallbladder and discuss under the light of current literature because of the rarity of these conditions. KEYWORDS: Gallbladder; Adenomyomatosis; Fundal variant. PMID- 21629531 TI - Oral and cutaneous melanoma: similarities and differences. AB - Melanomas are malignant lesions stemming from the disorganized proliferation of melanocytes. This condition is more common on skin, but may also be detected in mucosa, such as in the oral cavity. The aim of the present study was to report similarities and differences between oral and cutaneous melanoma. KEYWORDS: Melanoma; Skin; Mouth; Diagnosis. PMID- 21629532 TI - Long-term Safety of Testosterone and Growth Hormone Supplementation: A Retrospective Study of Metabolic, Cardiovascular, and Oncologic Outcomes. AB - BACKGROUND: Clinical research into the effects of hormonal supplementation has tended to focus on beneficial changes in anthropometric measures. There are fewer data on long-term safety with extended hormonal supplementation. METHODS: As part of a retrospective database survey, clinical outcomes were tabulated among patients who received at least 1 year of testosterone and/or growth hormone (GH) supplementation. In patients who were treated for at least 2 years, changes in markers of glucose and lipid metabolism were analyzed with and without concomitant use of oral hypoglycemics and statins. RESULTS: In 263 patients (mean age 56) treated for at least 2 years, the only statistically significant effect on markers of glucose metabolism was an increase in glycated hemoglobin (still within normal limits) in patients receiving GH alone or in combination with testosterone but without oral hypoglycemics; with or without hypoglycemics, insulin levels showed no significant change. The only significant effects on markers of lipid metabolism were decreases in total cholesterol and low-density lipoprotein (LDL) in patients receiving combined testosterone and GH without statins. Decreases in LDL were significant in both the statin and non-statin groups; decreases in triglycerides were significant only in the statin group. In 531 patients treated for at least 1 year (mean age 54), the overall incidence of adverse clinical outcomes (prostate disease, diabetes, cardiovascular disease, cancer) was 1.3%. CONCLUSIONS: In this retrospective survey, extended testosterone and/or GH supplementation did not adversely affect metabolic markers or clinical outcomes. KEYWORDS: Safety; Testosterone; Growth hormone; Supplementation. PMID- 21629533 TI - An Evaluation of Holiday Weight Gain Among Elementary-aged Children. AB - BACKGROUND: Previous studies suggest adults gain extra weight during the holiday season, however, few studies have been done with children during this time. The purpose of this study was to evaluate gains in growth among elementary children, and compare differences by gender and weight status. METHODS: Childrens (n = 90) height and weight were measured before and after their holiday break. Height, weight and body mass index (BMI) and body mass index-percentiles (BMI-%) were evaluated and compared by groups using repeated measures ANCOVAs. RESULTS: On average, children grew 0.82 cm (0.32 in), and gained 0.56 kg (1.2 lbs) and 0.28 BMI units, however the average BMI-% slightly decreased by 0.4%. Overweight and obese children gained significantly more weight, BMI units and BMI-% units compared with normal weight children. CONCLUSIONS: This study supports that the holiday period may be an important time to target children, especially those who are already overweight and obese. KEYWORDS: Holiday weight gain; Childhood obesity. PMID- 21629534 TI - Hydroscopic properties of organic objects that may present as aural foreign bodies. AB - BACKGROUND: Organic foreign bodies swell when irrigated with water, potentially making extraction more difficult. As the degree and rate of swelling of different types of organic foreign bodies has not been established, we aimed to analyze the hydroscopic properties of different organic foreign bodies in body temperature water. METHODS: Dry kidney beans, brown beans, peas, popcorn kernels, and dried fruits were soaked in a body temperature (37C) water bath. Volume of these organic materials was measured hourly to 8 hours, then at 12, 16, 24, 28, 36 and 48 hours. RESULTS: All dried fruits and beans increased in volume over time. The volume increase from baseline at 6 hours was between 43% (popcorn kernels) and 383% (kidney beans). Peas, popcorn, and raisins did not increase volume further after 6 hours. Kidney and brown beans had the greatest increase in volume overall (1268% and 482% respectively), and the greatest continued increase after 24 hours. CONCLUSIONS: Many organic substances that frequently present as aural foreign bodies may swell enough in water to lodge tightly in the ear canal. Typical popcorn kernels and dried peas will not swell sufficiently to lodge tightly in the ear canal of a typical child one year or older. A retained organic foreign body in a moist ear canal may cause inflammation until the foreign body can be removed. These risks may be offset by the advantages of successful removal with irrigation. KEYWORDS: Foreign body; Irrigation; Organic; Ear; Hydroscopic; Procedure; Removal. PMID- 21629535 TI - Three-year experience with rectal prolapse patients. AB - BACKGROUND: Rectal prolapse (RP) is a rare condition characterized by rectums protrusion through the anus with all of its layers. RP is a condition deteriorating the quality of life. Although more than 100 surgical procedures were described so far for the treatment of RP, the ideal treatment method still remains unclear. In this study, demographical data and clinical results of 13 patients who were treated at our clinic for RP for a period of 3 years were retrospectively studied, with the aim of comparing with the results of other repair methods mentioned in the literature. METHODS: Total of 13 patients admitted to the general surgery unit and the emergency units between January 2008 and December 2010 were included in the study. All of the cases were treated by modified Notoras technique using various synthetic materials. RESULTS: Of the patients, 8 were male, and 5 were female. Average age was 45.6 years (range: 23 - 79 years), and the average hospitalization time was 11.3 days (range: 3 - 19 days), with the symptom time being an average of 12 years (range: 1 - 30 years). All patients having complaints described mass prolapsing from the anal canal during defecation, rectal pain, and constipation. Six of our patients also had complaints of rectal bleeding. Average follow-up time was 24 months. No recurrence and mortality were monitored in patients who were followed. CONCLUSIONS: The main purposes in the surgical treatment of RP were to control the prolapse, and to achieve continence and remedy constipation. We believe that the modified Notoras technique made using synthetic materials the most suitable one compared to other rectopexy methods in the treatment of RP because it is safe and easily applicable. KEYWORDS: Rectal prolapse; Rectopexy; Modified Notoras technique. PMID- 21629536 TI - Parenchymal thoracic splenosis: history and nuclear imaging without invasive procedures may provide diagnosis. AB - Splenosis is a rare finding of ectopic splenic tissue found within the thoracic cavity, abdomen or peritoneal cavity. Most cases occur in the abdomen and the thoracic location is a comparatively rare finding. In thoracic splenosis the splenic tissue most often grows in the form of a nodule and the autotransplantation is usually caused by a previous operation and/or most commonly a penetrating or blunt trauma to the thoracoabdominal region, resulting in splenic rupture and in some cases left diaphragmatic tear. In majority of the cases the patients are asymptomatic and are incidentally diagnosed with left hemithorax pulmonary lesions found via chest radiography or thoracic computed tomography. We present a 45-year-old Caucasian male who was incidentally diagnosed with parenchymal thoracic splenosis secondary to a gunshot wound to the abdomen 13 years ago that resulted in distal pancreatectomy, splenectomy and gastrorrhaphy. In this case report we will briefly discuss the current updates in the literature regarding thoracic splenosis, and highlight the fact that the findings raise the suspicion of malignancy requiring numerous investigations yet early recognition of thoracic splenosis can prevent unnecessary tests and procedures. Preoperative diagnosis of splenosis should be made with the use of nuclear imaging studies such as the (99m)Tc heat-damaged erythrocyte study rather than computed tomography-guided biopsy or invasive surgery. KEYWORDS: Thoracic splenosis; Computed tomography; Ppancreatectomy; Splenectomy; Gastrorrhaphy. PMID- 21629537 TI - Cutaneous granular cell tumor of the breast: a clinical diagnostic pitfall. AB - We report the clinical-morphological study of a granular cell tumor in dermal/hypodermal junction and subcutaneous fat left breast of an 83-year-old woman with a family history of breast carcinoma. Mammography study showed a spiculated lesion in the lower inner quadrant with suspicion of malignancy. The results of fine needle puncture-aspiration were inconclusive. Subsequent tumorectomy revealed a poorly-defined indurated lesion of 1.1 x 0.7 cm. The histopathology study showed a proliferation of cells with ample and granular cytoplasm that were positive for S100, CD 68 and inhibin and negative for hormonal receptors. We present a benign lesion that clinically reproduces a breast carcinoma. KEYWORDS: Granular cell tumor; Breast; Differential diagnosis; Cutaneous. PMID- 21629538 TI - Upper respiratory tract symptoms, renal involvement and vasculitis: a case report and review of wegener granulomatosis. AB - Wegeners Granulomatosis is a condition associated with systemic vasculitis which can present with upper respiratory tract symptoms initially. On September 2001, a 15-year-old girl presented with symptoms of nasal block for 3 weeks. She later developed joint pains and worsening renal status requiring dialysis. A renal biopsy was performed which showed pauci-immune cresentric glomerulonephritis. Her cANCA levels were positive. She was treated with oral cyclophosphamide and steroids and later responded. KEYWORDS: Wegener granulomatosis; Young girl; Cyclophophamide; cANCA. PMID- 21629539 TI - Wilms tumor in a 37-year-old. AB - Wilms tumor is rare in adults. Though the approach to diagnosis and treatment of adult Wilms tumor (AWT) is closely modeled on recommendations for childhood Wilms tumor, views differ on how aggressive the treatment should be. We report a case of a 37-year-old with Stage III favorable histology AWT. A radical nephrectomy was performed and the patient was due for chemotherapy. Recent advances, controversies and current recommendations in the treatment of AWT are discussed. KEYWORDS: Adult; Wilms tumor; Kidney. PMID- 21629540 TI - A case of spontaneous intestinal perforation in osteogenesis imperfecta. AB - A 51-year-old male with known osteogenesis imperfecta (OI) (type 1) presented with symptoms and signs of infective endocarditis. Transthoracic echocardiography showed chordal rupture and free mitral regurgitation, resulting in an emergency mitral valve repair. The surgical procedure was largely uneventful but subsequent clinical course on the intensive care unit was complicated by bowel perforation requiring two laparatomies for a colonic resection and loop ileostomy formation. Histology of the excised tissue demonstrated absent musculature with no evidence of ischemia. Spontaneous non-ischemic bowel perforation as a complication of osteogenesis imperfecta is to date unreported. Our case highlights the need for a high index of suspicion of non-ischemic bowel perforation in patients with connective tissue disorders. KEYWORDS: Osteogenesis imperfecta; Bowel perforation; Collagen; Non-ischemic; Connective tissue disorders; Pathogenesis; Collagen vascular disorder; Acute abdomen. PMID- 21629541 TI - High Total Bilirubin as a Protective Factor for Diabetes Mellitus: An Analysis of NHANES Data From 1999 - 2006. AB - BACKGROUND: Diabetes Mellitus (DM) is a rampantly growing epidemic in the United States, affecting nearly 10% of the adult population. Studies have shown that higher levels of Total Bilirubin (TBili) convey a protective effect with regard to cardiovascular risk. In this study, we will examine the relationship between TBili level and prevalence of DM to discern whether a similar relationship exists. METHODS: The National Health and Nutrition Examination Survey (NHANES) is a comprehensive survey performed regularly to evaluate the overall health and nutrition status of the United States population. For the purpose of this study, we combined NHANES data collected between 1999 and 2006. Totally 15,876 eligible participants were selected after excluding all patients younger than twenty years, those with a history of abnormal liver function tests, or those who disclosed a history of liver disease. The data collected on these individuals was adjusted for demographic characteristics, as well as risk factors for DM, and was analyzed via multivariate logistic regression, using SAS proc survey methodology. RESULTS: After age adjustment, increased TBili was associated with 26% reduction in diabetes risk (OR 0.74, 95% CI 0.64 - 0.88). Multivariate analysis, adjusting for all diabetes risk factors assessed, confirmed this association (OR 0.80, 95% CI 0.67 - 0.95). CONCLUSIONS: Our results show that a higher level of serum TBili is associated with odds of having a lower incidence of DM. This finding supports the hypothesis that the antioxidant nature of TBili, demonstrating a protective effect with regard to the risk of stroke, atherosclerosis, and vasculitis in prior research, also extends to DM risk. Furthermore, research has shown that higher levels of TBili increase glucose mobilization into the cells, leading to more efficient, biologic glucose utilization. There is no doubt that the beneficial effect of TBili is multifactorial; thus further investigation is warranted. KEYWORDS: Bilirubin; Diabetes; Antioxidant; Protective. PMID- 21629542 TI - Exercise training prevents endometrial hyperplasia and biomarkers for endometrial cancer in rat model of type 1 diabetes. AB - BACKGROUND: Endometrial cancer is one of the most common types of gynecologic cancers. The ability of exercise to reduce the risk of endometrial cancer in women with type 2 diabetes has been established, but no studies have examined this link in type 1 diabetes.A randomized, controlled animal study was designed using a standard rat model of type 1 diabetes. The goal of this study was to investigate the ability of exercise to prevent increased levels of endometrial cancer biomarkers, estrogen receptor (ERalpha) and p16, and endometrial hyperplasia associated with diabetes. METHODS: FORTY FEMALE RATS WERE RANDOMIZED INTO FOUR GROUPS: sedentary control, exercise control, sedentary or exercised diabetic. Diabetes was induced by alloxan injection. A 4-week treadmill training program was initiated with the development of diabetes. Endometrial tissues were evaluated for hyperplasia and ERalpha and p16 levels and subcellular localization using microscopy. RESULTS: Severe diabetes lead to hyperplasia in the endometrial tissue in 70% of sedentary diabetic rats. Exercise-trained diabetic rats and the non-diabetic rats displayed no hyperplasia. The expression of ERalpha increased significantly (p < 0.02) while the expression level of p16 decreased significantly (p < 0.04) in the diabetic sedentary group compared to the non diabetic groups. Exercise training led to a reversal in the percentage of p16 and ERalpha positive cells in diabetic rats. CONCLUSIONS: Severe diabetes leads to hyperplasia of the endometrial tissue and increased ERalpha levels and decreased p16 levels in rats, which can be prevented with aerobic exercise. KEYWORDS: Diabetes; Estrogen receptor alpha; P16; Endometrial hyperplasia; Endometrial cancer; Exercise. PMID- 21629543 TI - Inappropriate Use of Intravenous PPI for Stress Ulcer Prophylaxis in an Inner City Community Hospital. AB - BACKGROUND: Proton pump inhibitors (PPIs) are used for the treatment and prophylaxis of variety of acid peptic conditions including stress ulcers. There has been a persistent practice of their inappropriate use for stress ulcer prophylaxis. Purpose of our study was to measure the inappropriate use of Intravenous Proton Pump Inhibitors for stress ulcer prophylaxis and to estimate the financial burden. METHODS: We carried out a retrospective, analytic study from July 2008 to June 2009 in internal medicine department. Hospital pharmacy records were used to identify all patients who received IV PPI during hospital stay. Seventy-five percent of records were randomly chosen (n=1104). PPI application was defined as indicated according to AGA guidelines. RESULTS: Intravenous proton pump inhibitor (IV PPI) was prescribed for 68.5% of patients without any proper indication. The estimated cost of medication for inappropriate IV PPIS use during the study year was 18337 USD. CONCLUSIONS: A more rational use of PPI will have better impact on health care cost and is likely to add to patient safety. KEYWORDS: Inappropriate use of PPI; Stress ulcer prophylaxis; Healthcare cost. PMID- 21629545 TI - Unusual case of acute intestinal obstruction. AB - Chronic NSAID use can cause diaphragm like strictures within the bowel leading to intestinal obstruction. This rare entity is called diaphragm disease. Preoperative diagnosis is extremely difficult. NSAID related injury should be considered in patient with intestinal obstruction of unclear etiology. We hereby present an interesting case of a 46 years old lady with significant history of long term naproxen use, who presented with nausea, vomiting and abdominal pain. She was diagnosed with acute intestinal obstruction. She received a trial of conservative management which was unsuccessful. Exploratory laparotomy with segmental resection of ileum was done subsequently. Histopathology showed mucosal ulceration, inflammation and focal sub mucosal fibrosis consistent with diaphragm disease. Patient recovered well in the post operative period. KEYWORDS: NSAIDS; Acute Intestinal Obstruction; Diaphragm Disease. PMID- 21629544 TI - Adenosine Deaminase Levels in CSF of Tuberculous Meningitis Patients. AB - BACKGROUND: Tuberculosis kills five lakh patients in India every year, out of which 7-12 % are with meningeal involvement. Delay in its diagnosis and in initiation of treatment results in poor prognosis and sequlae in up to 25% of cases. The aim of the present study is to look for a simple, rapid, cost effective, non-invasive and fairly specific test in differentiating tubercular etiology from other causes. METHODS: Forty patients between the age of 6 - 24 months attending hospital with symptoms and signs of meningitis were selected and divided into two groups: tubercular and non-tubercular, depending upon the accepted criteria. CSF was drawn and ADA estimated. RESULTS: Out of 19 tubercular patients, 18 had CSF ADA at or above the cutoff value while one had below. Out of 21 non-tuberculous patients, two had ADA levels at or above the cutoff value while 19 had below this value. Results of our study indicate that ADA level estimation in CSF is not only of considerable value in the diagnosis of TBM, CSF ADA level 10 U/L as a cutoff value exhibited 94.73% sensitivity and 90.47% specificity in differentiating tuberculous from non-tuberculous meningitis; it also has 90.00% positive predictive value and 95.00% negative predictive value. CONCLUSIONS: It can be concluded that ADA estimation in CSF is not only simple, inexpensive and rapid but also fairly specific method for making a diagnosis of tuberculous etiology in TBM, especially when there is a dilemma of differentiating the tuberculous etiology from non-tuberculous ones. For this reason ADA estimation in TBM may find a place as a routine investigation. KEYWORDS: Cerebrospinal fluid; Adenosine deaminase; Tuberculous meningitis. PMID- 21629546 TI - Hypereosinophilia: a diagnostic dilemma. AB - A case of hypereosinophilia is presented. The case illustrates the complexity of the diagnostic processes in certain conditions like hypereosinophilia. KEYWORDS: Hypereosinophilia; Myocarditis; Stroke. PMID- 21629547 TI - Apocrine fibroadenoma of the perianal region associated with perianal fistula. AB - A case of a 45 year-old woman who presented with a perianal fistula was reported. Histologically, the excised lesion showed features of apocrine fibroadenoma. In addition, the lesion had both glandular and stromal growth patterns and active chronic inflammation in the background. Malignant or benign counterparts of these types of lesions are widely described in literature. However fistula does not usually accompany and is very rare. Therefore, it is crucial for the pathologists and clinicians to be aware of such lesions. The relationship between apocrine fibroadenoma and perianal region was also discussed. KEYWORDS: Apocrin fibroadenoma; Perianal region; Fistula. PMID- 21629548 TI - Ninety-seven Years Old Hispanic Male With Mantle Cell Lymphoma. AB - Treating geriatric patients can be difficult. Patients often present with subtle or atypical symptoms, and end-of-life care is rarely neatly packaged. We present a case of a 97 years old Hispanic male with multiple medical problems who was diagnosed with mantle cell lymphoma after thoracentesis from what was thought to be a para-pneumonic effusion. An oncology evaluation determined that he was not a candidate for aggressive treatment. There is no evidence of curative treatment for mantle cell lymphoma. To complicate the decision making process further, the patients primary caregiver requested specifically that the health care team withholds the diagnosis from the patient. We discuss the inherent difficulties associated with end-of-life care. KEYWORDS: Geriatrics; End-of-life care; Mantle cell lymphoma; Hispanic. PMID- 21629549 TI - The Journal of Chiropractic Medicine and selected specialties in the chiropractic profession. AB - This article presents a brief description of the specialty chiropractic groups that are affiliated with the Journal of Chiropractic Medicine: American Academy of Chiropractic Physicians (AACP), The American Chiropractic Board of Sports Physicians (ACBSP), The College on Forensic Sciences (CFS), and The International Academy of Chiropractic Neurology (IACN). PMID- 21629550 TI - Assessment of patients with neck pain: a review of definitions, selection criteria, and measurement tools. AB - OBJECTIVE: The purpose of this literature review was to synthesize the existing literature on various definitions, classifications, selection criteria, and outcome measures used in different studies in patients with neck pain. METHODS: A literature search of MEDLINE and CINAHL through September 2008 was performed to gather articles on the reliability, validity, and utility of a wide variety of outcome measurements for neck pain. RESULTS: Different types of definitions appear in the literature based on anatomical location, etiology, severity, and duration of symptoms. Classifications according to severity and duration of pain and the establishment of selection criteria seem to play a crucial role in study designs and in clinical settings to ensure homogeneous groups and effective interventions. A series of objective tests and subjective self-report measures are useful in assessing physical abilities, pain, functional ability, psychosocial well-being, general health status, and quality of life in patients with neck pain. Self-administered questionnaires are commonly used in clinical practice and research projects. CONCLUSIONS: Because of multidimensionality of chronic neck pain, more than just one index may be needed to gain a complete health profile of the patient with neck pain. The instruments chosen should be reliable, valid, and able to evaluate the effects of treatment. PMID- 21629551 TI - Inter- and intraexaminer reliability of the Blair protractoview method: examination of a chiropractic radiographic technique. AB - OBJECTIVE: The purpose of this study was to evaluate the inter- and intraexaminer reliability of the Blair protractoview radiographic method. METHODS: This retrospective study evaluated 25 participants attending a Blair technique seminar. Participants included chiropractic students and doctors of chiropractic with more than 11 years of experience. Participants evaluated 100 Blair protractoview radiographs (oblique nasium). A kappa analysis was used to determine the inter- and intraexaminer reliability because of the nominal categorical value of the variables. For the interexaminer reliability, a kappa score was given for each examiner combination. The scores were then averaged to give the total interexaminer reliability. RESULTS: The overall interexaminer reliability showed substantial reliability at 0.62. Within-group kappa values were as follows: no certification = 0.61, proficiency = 0.66, primary level = 0.61, and advanced level = 0.74. The overall intraexaminer reliability showed outstanding reliability at 0.81. Within-group kappa values were as follows: no certification = 0.76, proficiency = 0.84, primary level = 0.82, and advanced level = 0.92. All kappa values had a P value < .001. CONCLUSION: The participants in this study showed good inter- and intraexaminer reliability using the Blair protractoview radiographic method. PMID- 21629552 TI - An atypical case of nephrolithiasis with transient remission of symptoms following spinal manipulation. AB - OBJECTIVE: Nephrolithiasis is a common condition with symptoms similar to common mechanical lesions of the lumbar spine and pelvis. The purpose of this report is to outline a case of nephrolithiasis that closely mimicked sacroiliac joint syndrome in subjective report, objective findings, and reduction of symptoms with spinal manipulation. CLINICAL FEATURES: A 41-year-old obese male patient with mild pain over the left posterior sacroiliac joint, penile paresthesia, and the penile sensation of urinary urgency presented for chiropractic care. Subjective history and objective evaluation suggested sacroiliac joint syndrome. INTERVENTION AND OUTCOME: A trial of conservative management including spinal manipulation was initiated. Following each treatment, the patient reported temporary relief of all symptoms (4 hours to 2 days). After unsuccessful permanent resolution of symptoms, a urinalysis was performed; and a follow-up computerized tomography scan revealed a large renal calculus obstructing the left ureter. Laser lithotripsy produced obliteration of the stone and complete resolution of symptoms. CONCLUSIONS: This report outlines the potential overlap of symptoms of visceral and somatic lesions in both presentation and response to care. In this case, a favorable response to spinal manipulation masked the most likely underlying symptom generator. This encounter demonstrates the potential need for further clinical examination in the instance of the unresponsive mechanical lesion. This report also supports the need for future research into spinal manipulation as a possible adjunct for visceral pain management. PMID- 21629553 TI - Inferior lumbar triangle hernia as a rarely reported cause of low back pain: a report of 4 cases. AB - OBJECTIVE: Lumbar triangle hernias are rarely reported causes of low back pain. We describe the symptoms, signs, and anatomical location of 2 possible defects in the posterior abdominal wall where lumbar hernias may appear. The clinical diagnosis was challenging, and advanced imaging failed to initially uncover the conditions. CLINICAL FEATURES: We report 4 patients with spontaneous inferior lumbar triangle hernias (Petit triangle hernias) initially presenting to a primary care clinic with the primary complaint of low back pain. INTERVENTION AND OUTCOMES: Thorough histories and examinations led to successful outcomes. All 4 patients were operated on to correct the defect. No recurrence has occurred. CONCLUSIONS: Anatomical knowledge and clinical acumen led to correct diagnosis of these rare lumbar hernias. This information should help both medical and chiropractic clinicians detect these conditions, and aid in appropriate management. PMID- 21629554 TI - Management of a 59-year-old female patient with adult degenerative scoliosis using manipulation under anesthesia. AB - OBJECTIVE: Manipulation under anesthesia (MUA) is an outpatient procedure that is performed to restore normal joint kinematics and musculoskeletal function. This article presents a case of a patient with idiopathic lumbar degenerative scoliosis who developed intractable pain as an adult and reports on the outcomes following a trial of MUA. CLINICAL FEATURES: A 59-year-old female patient presented to a chiropractic office with primary subjective symptoms of lower back and bilateral hip pain. Numerical pain rating scores were reported at 8 of 10 for the lower back and 9 of 10 for the sacroiliac joint/gluteal region. A disability score using a functional rating index demonstrated a score of 26 of 40 (or 64% disability). Over the preceding 5 years, the patient had tried a number of conservative therapies to relieve her pain without success. INTERVENTION AND OUTCOME: The patient was evaluated for MUA. The patient was scheduled for a serial MUA over 3 days. Numerical pain rating scores 8 weeks after the MUA were 1 of 10 for the lower back and 3 of 10 for the sacroiliac joint. Her disability rating decreased to 11 of 40 (28%). Radiological improvements were also observed. These outcomes were maintained at 6-month follow-up. CONCLUSION: Pain, functional, and radiographic outcomes demonstrated improvements immediately following treatment for this patient. PMID- 21629555 TI - Triad of spinal pain, spinal joint dysfunction, and extremity pain in 4 pediatric cases of "Wii-itis": a 21st century pediatric condition. AB - OBJECTIVE: This article describes 4 pediatric cases of overuse injuries related to playing Nintendo Wii (Nintendo, Redmond, WA). A brief discussion is also presented regarding other 21st century problems found in the literature, such as problems associated with playing the Nintendo DS portable electronic device, text messaging, and Blackberry (Research in Motion, Waterloo, Ontario) thumb. CLINICAL FEATURES: Four pediatric patients, ranging from 3 to 9 years old, who had injuries causally related to what has been described in the literature as "Wii itis" (spinal pain, spinal joint dysfunction [chiropractic subluxation], and related extremity pain), presented to a chiropractic clinic. INTERVENTION AND OUTCOMES: Each of the 4 pediatric cases was evaluated and managed using chiropractic techniques. All patients successfully had their complaints resolve with 1 chiropractic visit. CONCLUSION: Children in the new era of portable electronic devices are presenting to chiropractic offices with a set of symptoms directly related to overuse or repetitive strain from prolonged play on these systems. PMID- 21629556 TI - Upper cervical chiropractic care for a 25-year-old woman with myoclonic seizures. AB - OBJECTIVE: The purpose of this case report is to describe the chiropractic management using upper cervical techniques of a 25-year-old woman diagnosed with juvenile myoclonic epilepsy (JME). CLINICAL FEATURES: A 25-year-old woman had a history of JME, which was diagnosed at the age of 14 years. Her seizure episodes began shortly after trauma to her cervical spine and the onset of menarche. INTERVENTION AND OUTCOME: After case history and physical examination, the patient received high-velocity, low-amplitude chiropractic spinal manipulation to her upper cervical spine using the Blair upper cervical chiropractic technique protocol. There was improvement in her seizure episodes and menstrual cycles following 12 weeks of chiropractic care. CONCLUSION: This case study demonstrated improvement in a young woman with a seizure disorder after she received upper cervical chiropractic manipulation. This case suggests the need for more rigorous research to examine how upper cervical chiropractic techniques may provide therapeutic benefit to patients with seizure disorders. PMID- 21629557 TI - The relationship between pelvic torsion and anatomical leg length inequality: a review of the literature. PMID- 21629559 TI - The American Chiropractic Board of Sports Physicians wishes to congratulate the following doctors for receiving their DACBSP and CCSP certifications in 2009. PMID- 21629558 TI - The relationship between pelvic torsion and anatomical leg length inequality: a review of the literature. PMID- 21629560 TI - Structural and dynamic properties of linker histone H1 binding to DNA. AB - Found in all eukaryotic cells, linker histones H1 are known to bind to and rearrange nucleosomal linker DNA. In vitro, the fundamental nature of H1/DNA interactions has attracted wide interest among research communities-from biologists to physicists. Hence, H1/DNA binding processes and structural and dynamical information about these self-assemblies are of broad importance. Targeting a quantitative understanding of H1 induced DNA compaction mechanisms, our strategy is based on using small-angle x-ray microdiffraction in combination with microfluidics. The usage of microfluidic hydrodynamic focusing devices facilitates a microscale control of these self-assembly processes, which cannot be achieved using conventional bulk setups. In addition, the method enables time resolved access to structure formation in situ, in particular, to transient intermediate states. The observed time dependent structure evolution shows that the H1/DNA interaction can be described as a two-step process: an initial unspecific binding of H1 to DNA is followed by a rearrangement of molecules within the formed assemblies. The second step is most likely induced by interactions between the DNA and the H1's charged side chains. This leads to an increase in lattice spacing within the DNA/protein assembly and induces a decrease in the correlation length of the mesophases, probably due to a local bending of the DNA. PMID- 21629561 TI - Use of a virtual wall valve in polydimethylsiloxane microfluidic devices for bioanalytical applications. AB - A simple method for micromanipulation of liquids and/or small groups of cells is presented in this study. Microfabricated sieving structures composed of PDMS (polydimethylsiloxane) were used to segregate aqueous solutions. This microfluidic valving scheme was an application of Cassie-Baxter wetting and was termed "virtual walls" as a nonsolid barrier exists at an air/water interface. The manipulation of the virtual-air-wall valve was accomplished by controlling the strength of surface-tension and hydrostatic-pressure forces. Virtual walls with a range of feature sizes were designed and characterized by monitoring air and water displacement in response to hydrostatic pressure. Thresholds for the virtual-air-wall valves to be turned on or off were quantified. The walls could also be formed or dissipated by the focused microbeam of a pulsed laser. As an illustration of the virtual wall utility, a series of microfluidic applications were demonstrated. First, the capability of virtual walls to temporarily segregate liquids was integrated into a device utilized to establish a chemical gradient. In a second application, the arraying of nonadherent cells within individual aqueous cavities created by the virtual walls was demonstrated. Individual cells were also released from the cavities on demand using a focused microbeam. The virtual walls were simple and easy-to-fabricate without the requirement for surface treatment or precision alignment, and should find usage in bioanalytical applications. PMID- 21629562 TI - CdTe quantum dots and polymer nanocomposites for x-ray scintillation and imaging. AB - Investigations are reported on the x-ray scintillation and imaging application of CdTe quantum dots (QDs) and their polymer nanocomposites. Aqueous CdTe QDs with emissions ranging between 510 and 680 nm were prepared and incorporated into polyvinyl alcohol or polymethyl methacrylate polymer matrices. The x-ray luminescent properties were evaluated and a resolution of 5 lines/mm was obtained from the nanocomposite films. Additionally, the fast decay time, nonafterglow, and superior spectral match to conventional charge coupled devices, show that CdTe QD nanocomposites have high promise for x-ray imaging applications. PMID- 21629563 TI - Face shear piezoelectric properties of relaxor-PbTiO(3) single crystals. AB - Poling relaxor-PbTiO(3) single crystals along pseudocubic [011] results in a macroscopic symmetry of mm2, enabling a large face shear d(36) in Zt+/-45 degrees cut crystals. In order to allow the determination of electrical properties by the resonance method, square samples are required. Using Pb(In(0.5)Nb(0.5))O(3) Pb(Mg(1/3)Nb(2/3))O(3)-PbTiO(3) crystals, piezoelectric d(36) coefficients were determined to be in the range of 2000-2500 pC/N, with electromechanical coupling factor k(36)~0.80-0.83. Mechanical quality factor Q~180 and ultralow frequency constant of ~500 Hz m were obtained. Together with the wide temperature usage range (up to ~110 degrees C) and high ac driving field stability (~5 kV/cm), such face shear crystals have a promising potential for ultralow-frequency transducer applications. PMID- 21629564 TI - A poly(vinylidene fluoride) composite with added self-passivated microaluminum and nanoaluminum particles for enhanced thermal conductivity. AB - A polymer composite was prepared by embedding fillers made of self-passivated aluminum particles in two kind of sizes, micrometer size and nanometer size with different volume proportions into polyvinylidene fluoride matrix. The thermal conductivity and dielectric properties of the composite were studied. The results showed that the thermal conductivity of composites was significantly increased to 3.258 W/mK when the volume proportion of micrometer size Al particles to nanometer size Al particles is at 20:1, also the relative permittivity was about 75.8 at 1 MHz. The effective simulation model values were in good accordance with experimental results. PMID- 21629565 TI - Coaxial atomic force microscope probes for imaging with dielectrophoresis. AB - We demonstrate atomic force microscope (AFM) imaging using dielectrophoresis (DEP) with coaxial probes. DEP provides force contrast allowing coaxial probes to image with enhanced spatial resolution. We model a coaxial probe as an electric dipole to provide analytic formulas for DEP between a dipole, dielectric spheres, and a dielectric substrate. AFM images taken of dielectric spheres with and without an applied electric field show the disappearance of artifacts when imaging with DEP. Quantitative agreement between our model and experiment shows that we are imaging with DEP. PMID- 21629566 TI - A novel mutation in SLC37A4 gene in a Sri Lankan boy with glycogen storage disease type Ib associated with very early onset neutropenia. PMID- 21629567 TI - Switching patients from preserved prostaglandin-analog monotherapy to preservative-free tafluprost. AB - PURPOSE: Efficacy, tolerability and safety of the novel preservative-free prostaglandin tafluprost 0.0015% were investigated for the treatment of patients with glaucoma or ocular hypertension in a clinical setting. PATIENTS AND METHODS: Data were collected in a non-interventional, prospective, multi-center, observational, open label study. 118 patients were treated with a prostaglandin analog (PGA) monotherapy (preserved formulations of latanoprost, travoprost or bimatoprost) prior to baseline. Intraocular pressure (IOP) readings were recorded for each eye at baseline (previous therapy), 4-6 weeks, and 12 weeks after changing medical treatment to preservative-free tafluprost once-daily. We analyzed the change in IOP over the study period for all patients as well as for a subgroup of patients with prior PGA monotherapy. Subjective symptoms and objective ocular signs were determined. Comfort was measured using a 4 step scale. All adverse events were recorded. Paired t-tests were conducted to compare IOP values at baseline to IOP values after treatment with tafluprost 0.0015%. Bowker's test of symmetry was used for statistical evaluation of changes of clinical signs (hyperemia). RESULTS: In total 118 patients were eligible for evaluation. In these patients with prior PGA monotherapy (n = 118) IOP decreased significantly from 16.2 +/- 4.3 mm Hg (95% CI: 0.55) at treated baseline to 14.8 +/- 3.2 mm Hg (95% CI: 0.43; P < 0.001) at final visit on tafluprost. In a subset of patients with prior latanoprost monotherapy (n = 68) mean IOP at baseline (+/ SD) was reduced from 16.2 +/- 4.6 mm Hg (95% CI: 0.77) 14.8 +/- 3.1 mm Hg at final visit (95% CI: 0.54, P < 0.001), in patients with prior travoprost monotherapy (n = 32) from 16.2 +/- 4.3 mm Hg (95% CI: 1.05) to 14.9 +/- 3.3 mm Hg (95% CI: 0.91; P < 0.05) and in patients with prior bimatoprost monotherapy (n = 18) from 16.4 +/- 3.5 mm Hg (95% CI: 1.14) to 15.0 +/- 3.3 mm Hg (95% CI: 1.14; P = 0.252). Both, objective clinical signs and subjective symptoms improved after changing medication to preservative-free tafluprost until final visit. The number of patients with moderate and severe hyperemia decreased from 51 (43.2%) at baseline to 2 (1.9%) at final visit. CONCLUSION: Preservative-free tafluprost 0.0015% was effective, well tolerated and safe. IOP was controlled effectively and ocular symptoms and clinical signs were improved after changing medication to a monotherapy with preservative-free tafluprost in patients previously treated with a preserved latanoprost, travoprost or bimatoprost monotherapy. PMID- 21629568 TI - Ocular iontophoresis of EGP-437 (dexamethasone phosphate) in dry eye patients: results of a randomized clinical trial. AB - PURPOSE: To assess safety and efficacy of EGP-437 (dexamethasone phosphate 40 mg/mL [DP]) in dry eye patients. METHODS: The study employed a prospective, single-center, double-masked design utilizing a Controlled Adverse Environment (CAE). Patients (n = 103) with confirmed signs and symptoms of dry eye syndrome were randomized into 1 of 3 iontophoresis treatment groups: 7.5 mA-min at 2.5 mA (DP 7.5, n = 41); 10.5 mA-min at 3.5 mA (DP 10.5, n = 37); or 10.5 mA-min at 3.5 mA (placebo, n = 25). Three CAE visits and 4 follow-up visits occurred over 3 weeks. Patients meeting enrollment criteria received iontophoresis in both eyes after the second CAE exposure (visit 3) and before the third CAE exposure (visit 5). Primary efficacy endpoints were corneal staining and ocular discomfort. Secondary endpoints included tear film break-up time, ocular protection index (OPI), and symptomatology. RESULTS: The DP 7.5 and DP 10.5 treatment groups showed statistically significant improvements in signs and symptoms of dry eye at various time points; however, the primary endpoints were not achieved. The DP 7.5 treatment group exhibited statistically significant improvements in corneal staining (when comparing the differences between study entry and exit, 3 weeks, P = 0.039), OPI (immediately following the second treatment, P = 0.048) and ocular discomfort at follow-up visits (a week after the first treatment, P = 0.032; 24 hours after the second treatment, P = 0.0032). Treatment-emergent adverse events (AEs) were experienced by 87% of patients and were consistent across all treatment groups. Most AEs were mild and no severe AEs were observed. CONCLUSION: Ocular iontophoresis of EGP-437 demonstrated statistically and clinically significant improvements in signs and symptoms of dry eye syndrome within a CAE model. PMID- 21629569 TI - Evaluation of spectral domain and time domain optical coherence tomography findings in toxoplasmic retinochoroiditis. AB - PURPOSE: To investigate spectral domain optical coherence tomography (SD-OCT) findings and compare them with time domain (TD)-OCT imaging of macula and retinochoroiditis lesions of patients with toxoplasmosis. DESIGN: Prospective cross-sectional study. METHODS: Ten eyes of 10 patients with active toxoplasmic retinochoroiditis were included. Morphologic features from the macula and retinochoroiditis lesions were obtained at baseline and at 6-week follow up. Scan acquisition protocols for TD-OCT included raster and radial lines through the retinochoroiditis lesion, fast macular, and a linear scan from the lesion to the fovea, whereas the acquisition protocols for SD-OCT also included horizontal volume scans at the lesion site and at the macula. Thickness measurements obtained by SD-OCT were analyzed. RESULTS: At baseline, macular serous retinal detachment was observed in five patients; two of them only seen by SD-OCT. Retinochoroidal lesions were 4260 MUm distant from the fovea on average (R = 681 7130) and this distance had an indirect correlation to the presence of macular detachment. Epiretinal membrane and vitreo-macular traction were also observed. The posterior hyaloid was not identified in four patients by TD-OCT and only in one by SD-OCT at baseline. Perilesional subretinal fluid was observed in two patients. The median retinal thickness significantly decreased at the retinochoroiditis lesion (P = 0.0004), and all the patients remained with disorganized retinal layers reflectivity at follow up. CONCLUSION: SD-OCT is a useful tool in the diagnosis of macular changes related with toxoplasmic retinochoroiditis. SD-OCT is superior in evaluating retinal changes associated with ocular toxoplasmosis. PMID- 21629570 TI - Ocular injuries from exploding glass-bottled Coca-Cola(r) drinks in Port Harcourt, Nigeria. AB - BACKGROUND: Eye injuries and subsequent loss of vision from the glass and caps of exploding pressurized bottled drinks have been well reported, and as a result most developed countries now use mainly plastic bottles. In Nigeria, however, most drinks are still sold in glass bottles and ocular injuries from this source are therefore not uncommon. AIM: To retrospectively analyze ocular injuries resulting from exploding glass-bottled Coca-Cola(r) and propose ways of eliminating such injuries in future. SETTING: Eye Clinic, University of Port Harcourt Teaching Hospital, Port Harcourt, Nigeria. MATERIALS AND METHODS: The medical records of all cases of ocular injury that presented at the Eye Clinic of the University of Port Harcourt Teaching Hospital over a 5-year period (January 2006 to December 2010) were retrieved and relevant data including age, sex, occupation, events surrounding bottle explosion, and type of ocular injury sustained were extracted. RESULTS: A total of 426 cases of ocular injuries was seen during the period under review. There were 335 (78.6%) males and 91 (21.4%) females. Six patients had ocular injury from exploding glass-bottled Coca Cola(r), giving an incidence of 1.4%. The presenting visual acuities (VA) were light perception (2 cases), counting fingers (2 cases), and 1 VA of 6/24 and 1 VA of 6/12. There were 4 (66.7%) cases of corneoscleral laceration with uveal prolapse and 1 case of total hyphema. CONCLUSION: Because pressurized glass bottles can explode with normal handling, legislation to ban the use of glass containers for bottling carbonated drinks will go a long way to reducing ocular morbidity from this source. Plastic bottles should be introduced as an alternative. PMID- 21629571 TI - Alteration of melatonin secretion in patients with type 2 diabetes and proliferative diabetic retinopathy. AB - BACKGROUND: The purpose of this study was to evaluate the dynamics of plasma melatonin secretion in patients with type 2 diabetes mellitus and diabetic retinopathy. METHODS: Plasma melatonin levels were measured by high-performance liquid chromatography in 56 patients. Patients were divided into a diabetic group (30 patients) and a nondiabetic group (26 patients). The diabetic group was divided further into a proliferative diabetic retinopathy (PDR) group (n = 14) and a nonproliferative diabetic retinopathy (NPDR) group (n = 16). Plasma melatonin levels obtained at midnight and 3 am were compared between the groups. RESULTS: Nighttime melatonin levels were significantly lower in the diabetic group than in the nondiabetic group (P < 0.03) and lower in the PDR group than in the nondiabetic and NPDR groups (P < 0.01 and P < 0.03, respectively), but no significant difference was found between the nondiabetic and NPDR groups. The daytime melatonin level did not significantly differ between the nondiabetic and diabetic groups or between the nondiabetic, NPDR, and PDR groups. CONCLUSION: The nighttime melatonin level is altered in patients with diabetes and PDR but not in diabetic patients without PDR. Although patients with PDR may have various dysfunctions that affect melatonin secretion more severely, advanced dysfunction of retinal light perception may cause altered melatonin secretion. Alteration of melatonin secretion may accelerate further occurrence of complications in diabetic patients. PMID- 21629572 TI - Performance of the PT100 noncontact tonometer in healthy eyes. AB - BACKGROUND: The purpose of this study was to assess the repeatability and reproducibility of the PT100 noncontact tonometer and to compare its consistency with the Goldmann applanation tonometer (GAT) in measuring intraocular pressure (IOP). METHODS: Triplicate IOP measurements were obtained on two separate occasions using the PT100 and GAT from randomly selected eyes in 66 healthy volunteers aged 22 +/- 1 years. The repeatability and reproducibility of each techniques was assessed. Agreement between the techniques was statistically quantified using intrasession repeatability for each technique as the basis for comparison. RESULTS: Both techniques returned equal IOP values in the first measurement session (15 +/- 3 mmHg). The second session showed a mean difference in average IOP (1 +/- 0.71). The 95% limits of agreement between the techniques were -5.2 to 5.5 mmHg and -4.0 to 4.7 mmHg (sessions 1 and 2, respectively). These mean differences were not statistically significant (P > 0.05, paired t test), with the PT100 underestimating IOP measurement by 1.00 mmHg. The mean intrasession IOP for GAT sessions 1 and 2 was 0 +/- 0.90 mmHg and 0.04 +/- 1.06 mmHg, respectively, and the corresponding mean IOP measurement difference for the PT100 was -0.06 +/- 0.96 and -0.39 +/- 0.94 mmHg (sessions 1 and 2, respectively; P > 0.05, paired t-test). Repeatability coefficients for the GAT IOP measurements were 1.8 mmHg and 2.1 mmHg for sessions 1 and 2, while the PT100 repeatability coefficient was 1.9 mmHg and 1.8 mmHg for sessions 1 and 2, respectively. The intrasession repeatability coefficient of both techniques for test-retest differences were within +/-5 mmHg. CONCLUSION: The PT100 noncontact tonometer produced greater repeatability than the GAT in assessment of IOP, whereas GAT resulted in more reproducible results. Both techniques showed a close level of agreement on comparison, with the PT100 underestimating IOP measurement by 1.0 mmHg only, although this was not clinically or statistically significant. Of importance is that the IOP measurements using these techniques could be interchangeable in the IOP range studied here. PMID- 21629573 TI - Novel ocular antihypertensive compounds in clinical trials. AB - INTRODUCTION: Glaucoma is a multifactorial disease characterized by progressive optic nerve injury and visual field defects. Elevated intraocular pressure (IOP) is the most widely recognized risk factor for the onset and progression of open angle glaucoma, and IOP-lowering medications comprise the primary treatment strategy. IOP elevation in glaucoma is associated with diminished or obstructed aqueous humor outflow. Pharmacotherapy reduces IOP by suppressing aqueous inflow and/or increasing aqueous outflow. PURPOSE: This review focuses on novel non-FDA approved ocular antihypertensive compounds being investigated for IOP reduction in ocular hypertensive and glaucoma patients in active clinical trials within approximately the past 2 years. METHODS: The mode of IOP reduction, pharmacology, efficacy, and safety of these new agents were assessed. Relevant drug efficacy and safety trials were identified from searches of various scientific literature databases and clinical trial registries. Compounds with no specified drug class, insufficient background information, reformulations, and fixed-combinations of marketed drugs were not considered. RESULTS: The investigational agents identified comprise those that act on the same targets of established drug classes approved by the FDA (ie, prostaglandin analogs and beta-adrenergic blockers) as well as agents belonging to novel drug classes with unique mechanisms of action. Novel targets and compounds evaluated in clinical trials include an actin polymerization inhibitor (ie, latrunculin), Rho-associated protein kinase inhibitors, adenosine receptor analogs, an angiotensin II type 1 receptor antagonist, cannabinoid receptor agonists, and a serotonin receptor antagonist. CONCLUSION: The clinical value of novel compounds for the treatment of glaucoma will depend ultimately on demonstrating favorable efficacy and benefit-to-risk ratios relative to currently approved prostaglandin analogs and beta-blockers and/or having complementary modes of action. PMID- 21629574 TI - Influence of ischemia on visual function in patients with branch retinal vein occlusion and macular edema. AB - Visual function and retinal morphology were investigated to elucidate the influence of ischemia in patients with branch retinal vein occlusion (BRVO) and macular edema. In 41 consecutive patients with BRVO aged 68.9 +/- 10.0 years (22 women and 19 men), the area of capillary nonperfusion was measured by fluorescein angiography. Retinal thickness and retinal volume were measured by optical coherence tomography, and mean retinal sensitivity was calculated for each of 9 macular subfields. Mean visual acuity and macular sensitivity within the central subfield were not significantly correlated with the nonperfused area. However, the macular sensitivity within the central 5 subfields and all 9 subfields showed significant negative correlations with the nonperfused area. Also, macular thickness and volume within all 9 subfields were significantly correlated with the nonperfused area. In conclusion, evaluation of both the fovea and the entire macular region may be important in patients with ischemic BRVO. PMID- 21629575 TI - Early-stage mucinous sweat gland adenocarcinoma of eyelid. AB - We present the findings of an early-stage primary mucinous sweat gland adenocarcinoma in the lower eyelid of a Japanese patient. The patient was a 73 year-old man who had had a nodule on the left lower eyelid for two years. He was referred to our hospital with a diagnosis of a swollen chalazion. The clinical and histopathological records were reviewed and the mass was excised. Histopathological examination revealed a mucinous sweat gland adenocarcinoma. Postoperative magnetic resonance imaging and positron emission tomography excluded systemic metastases. After the histopathological findings, a complete surgical excision of the margins of the adenocarcinoma was performed, with histopathological confirmation of negative margins. After the final histopathological examination, the patient was diagnosed with a primary mucinous sweat gland adenocarcinoma of the left eyelid. Six months after the surgery, no recurrence has been observed. Because the appearance of mucinous sweat gland adenocarcinoma of the eyelid is quite variable, the final diagnosis can only be made by histopathological examination. A complete surgical excision is recommended. PMID- 21629576 TI - Potential role of intravitreal human placental stem cell implants in inhibiting progression of diabetic retinopathy in type 2 diabetes: neuroprotective growth factors in the vitreous. AB - BACKGROUND: Intravitreal injection of human mesenchymal stem cells has been shown to be effective in slowing the progression of diabetic retinopathy in an animal model of chemically induced diabetes mellitus. We studied changes in growth factor levels released from human mesenchymal stem cells in the vitreous cavity as well as changes in growth factor levels in host retinal neurons following intravitreal injection. METHODS: Twenty-two Lewis rats were treated with an intravitreal human mesenchymal stem cell microinjection. Determination of neurotrophic factors released by human mesenchymal stem cells in the vitreous was carried out using real-time polymerase chain reaction. RESULTS: Detectable levels of neurotrophic factors were identified postoperatively in the vitreous of all rats. CONCLUSION: Increased intravitreal and retinal concentrations of neuroprotective growth factors in rats confirm the neuroprotective activity of human mesenchymal stem cells in diabetic retinopathy. PMID- 21629577 TI - Pharmacokinetics of intravitreal bevacizumab (Avastin(r)) in rabbits. AB - PURPOSE: To describe the pharmacokinetics of intravitreal bevacizumab (Avastin(r)) in rabbits. METHODS: The right eye of 20 rabbits was injected intravitreally with 1.25 mg/0.05 mL bevacizumab. Both eyes of four rabbits each time were enucleated at days 1, 3, 8, 15, and 29. Bevacizumab concentrations were measured in serum, aqueous humor, and vitreous. RESULTS: Maximum vitreous (406.25 MUg/mL) and aqueous humor (5.83 MUg/mL) concentrations of bevacizumab in the right eye were measured at day 1. Serum bevacizumab concentration peaked at day 8 (0.413 MUg/mL) and declined to 0.032 MUg/mL at 4 weeks. Half-life values in right vitreous, right aqueous humor, and serum were 6.61, 6.51, and 5.87 days, respectively. Concentration of bevacizumab in the vitreous of the noninjected eye peaked at day 8 (0.335 ng/mL) and declined to 0.218 ng/mL at 4 weeks. In the aqueous humor of the noninjected eye, maximum concentration of bevacizumab was achieved at day 8 (1.6125 ng/mL) and declined (to 0.11 ng/mL) at 4 weeks. CONCLUSION: The vitreous half-life of 1.25 mg/0.05 mL intravitreal bevacizumab was 6.61 days in this rabbit model. Maximum concentrations of bevacizumab were reached at day 1 in both vitreous and aqueous humor of the right eye and at day 8 in the serum. Very low concentrations of bevacizumab were measured in the fellow noninjected eye. PMID- 21629578 TI - Treatment of macular edema due to retinal vein occlusions. AB - Retinal vein occlusion (RVO) is a prevalent retinal vascular disease, second only to diabetic retinopathy. Previously there was no treatment for central retinal vein occlusion (CRVO) and patients were simply observed for the development of severe complications, generally resulting in poor visual outcomes. The only treatment for branch vein occlusion (BRVO) was grid laser photocoagulation, which reduces edema very slowly and provides benefit in some, but not all patients. Within the past year, clinical trials have demonstrated the effects of three new pharmacologic treatments, ranibizumab, triamcinolone acetonide, and dexamethasone implants. The benefit/risk ratio is best for intraocular injections of ranibizumab, making this first-line therapy for most patients with CRVO or BRVO, while intraocular steroids are likely to play adjunctive roles. Standard care for patients with RVO has changed and will continue to evolve as results with other new agents are revealed. PMID- 21629579 TI - Anterior segment changes following intravitreal bevacizumab injection for treatment of neovascular glaucoma. AB - BACKGROUND: The purpose of this study was to describe anterior segment changes in a prospective, interventional, noncomparative case series of patients with neovascular glaucoma secondary to proliferative diabetic retinopathy treated with intravitreal bevacizumab. METHODS: Five consecutive patients with neovascular glaucoma and a refractory, symptomatic elevation of intraocular pressure and pronounced anterior segment congestion received intravitreal bevacizumab 1.25 mg/0.05 mL. Follow-up examinations were performed at 4-16 weeks by the same specialists, with testing performed at hour 48, week 1, and months 1, 3, and 6 after intravitreal bevacizumab. RESULTS: We observed a significant difference (P = 0.021) between initial and mean neovascularization at three months in all the quadrants. At three months, median intraocular pressure was 19 +/- 5.38 (range 12 26) mmHg. In three of the five cases, diode laser cyclophotocoagulation was required, and in one case a trabeculectomy was performed. One patient showed complete synechial angle closure 48 hours after treatment which required cyclodestructive procedures to normalize intraocular pressure. CONCLUSION: Intravitreal bevacizumab achieves complete regression of neovascularization in neovascular glaucoma secondary to proliferative diabetic retinopathy, and this regression is stable when associated with treatment of the underlying disease and should be investigated more thoroughly as an adjunct in the management of neovascular glaucoma. PMID- 21629580 TI - Dietary Advice on Prescription: A novel approach to dietary counseling. AB - This article describes a novel approach to giving dietary advice, which is called "Dietary Advice on Prescription" (DAP; Matordning pa Recept [MoR] in Swedish). It is the same principle as prescription on medicine and "Physical Activity on Prescription" (PAP; Fysisk aktivitet pa Recept [FaR] in Swedish). The main idea is that a written prescription will strengthen the oral advice and emphasize certain aspects of the dietary recommendation. The DAP is on the brink of being tested in a planned study. PMID- 21629581 TI - Gambling: Electronic friends or a threat to one's health and personal development? AB - Gambling has become quite common in Sweden. The Swedish National Institute of Public Health (2010) has reported that about 70% of the Swedish population has gambled at least once during the last 12 months. Half of the population had gambled with money by 18 years of age and about 11% had gambled for the first time when they were 12-years-old or younger. In the report from the Swedish National Institute of Public Health, gambling problems are related to health problems and risky alcohol consumption. The highest problem rate is found among men aged 18 to 24; almost 1 in 10 had some gambling problems. The share of problem gamblers is found to be twice as high among the under-age gamblers as it is among the population as a whole. Young people gamble less but develop gambling problems to a larger extent than adult gamblers. In this article young people's gambling and their gambling careers are analysed as a natural part of their internalisation of other adult habits. PMID- 21629582 TI - Hunting for health, well-being, and quality of life. AB - Health, well-being, quality of life, and lifestyle are central concepts within health science, although generally accepted definitions are still lacking. Lifestyle can either be seen as an independent variable and the cause of unhealthy behaviour or as a dependent variable, which is affected by conditions in the society. In the first case, the attention is directed on each individual case: maintaining or improving health requires changes in lifestyle and living habits. In this perspective, diet and physical activity are important features for health promotion. In the second case the attention is rather directed on structural conditions in society, for example the food industry, the lunches for children at school, and the "fast food" industry should be influenced to protect human health. The structural perspective has, so far, received restricted impact when it concerns prevention and promotion of health. Processes of individualisation in the society have to an increasing extent viewed health as an affair for the individual. The benefits of physical activity, healthy food and beverage, social support, and joy are documented scientifically. In general, the trend towards increasing responsibility for one's lifestyle and health is positive, but might reinforce the inequality in health. With an even harder climate in society there might be a risk that individual health projects undermine the solidarity and the will to accept costs for medical treatment and care for people who risk their health through an unhealthy and risk-taking lifestyle. However, we argue that peoples' well-being and quality of life presupposes a society that stands up for all people. PMID- 21629583 TI - Computational models of reinforcement learning: the role of dopamine as a reward signal. AB - Reinforcement learning is ubiquitous. Unlike other forms of learning, it involves the processing of fast yet content-poor feedback information to correct assumptions about the nature of a task or of a set of stimuli. This feedback information is often delivered as generic rewards or punishments, and has little to do with the stimulus features to be learned. How can such low-content feedback lead to such an efficient learning paradigm? Through a review of existing neuro computational models of reinforcement learning, we suggest that the efficiency of this type of learning resides in the dynamic and synergistic cooperation of brain systems that use different levels of computations. The implementation of reward signals at the synaptic, cellular, network and system levels give the organism the necessary robustness, adaptability and processing speed required for evolutionary and behavioral success. PMID- 21629584 TI - A new interpretation of P300 responses upon analysis of coherences. AB - Previous studies on cognitive dynamics showed that oscillatory responses of P300 are composed of mainly delta and theta responses. In the present study, for the first time, the long-distance intra-hemispheric event related coherence (auditory oddball paradigm) and evoked coherence (simple sound) were compared in order to evaluate the effects of cognitive tasks on the long-distance coherences. Seventeen healthy subjects (8 female, 9 male) were included in the study. The coherence was analyzed for delta (1-3.5 Hz), theta (4-7.5 Hz) and alpha (8-13 Hz) frequency ranges for (F(3)-P(3), F(4)-P(4), F(3)-T(7), F(4)-T(8), F(3)-O(1,) F(4) O(2)) electrode pairs. The coherence to target responses were higher than the non target and simple auditory response coherence. This difference is significant for the delta coherence for both hemispheres and for theta coherences over the left hemisphere. The highest coherences were recorded at fronto-temporal locations for all frequency bands (delta, theta, alpha). Furthermore, fronto-parietal coherences were higher than the fronto-occipital coherences for all frequency bands (delta, theta, alpha).These results show that the fronto-temporal and fronto-parietal connections are most relevant for the identification of the target signal. This analysis open the way for a new interpretation of dynamic localization results during cognitive tasks. PMID- 21629585 TI - Electrophysiological correlates of listening effort: neurodynamical modeling and measurement. AB - An increased listing effort represents a major problem in humans with hearing impairment. Neurodiagnostic methods for an objective listening effort estimation might support hearing instrument fitting procedures. However the cognitive neurodynamics of listening effort is far from being understood and its neural correlates have not been identified yet. In this paper we analyze the cognitive neurodynamics of listening effort by using methods of forward neurophysical modeling and time-scale electroencephalographic neurodiagnostics. In particular, we present a forward neurophysical model for auditory late responses (ALRs) as large-scale listening effort correlates. Here endogenously driven top-down projections related to listening effort are mapped to corticothalamic feedback pathways which were analyzed for the selective attention neurodynamics before. We show that this model represents well the time-scale phase stability analysis of experimental electroencephalographic data from auditory discrimination paradigms. It is concluded that the proposed neurophysical and neuropsychological framework is appropriate for the analysis of listening effort and might help to develop objective electroencephalographic methods for its estimation in future. PMID- 21629586 TI - A method for the estimation of functional brain connectivity from time-series data. AB - A central issue in cognitive neuroscience is which cortical areas are involved in managing information processing in a cognitive task and to understand their temporal interactions. Since the transfer of information in the form of electrical activity from one cortical region will in turn evoke electrical activity in other regions, the analysis of temporal synchronization provides a tool to understand neuronal information processing between cortical regions. We adopt a method for revealing time-dependent functional connectivity. We apply statistical analyses of phases to recover the information flow and the functional connectivity between cortical regions for high temporal resolution data. We further develop an evaluation method for these techniques based on two kinds of model networks. These networks consist of coupled Rossler attractors or of coupled stochastic Ornstein-Uhlenbeck systems. The implemented time-dependent coupling includes uni- and bi-directional connectivities as well as time delayed feedback. The synchronization dynamics of these networks are analyzed using the mean phase coherence, based on averaging over phase-differences, and the general synchronization index. The latter is based on the Shannon entropy. The combination of these with a parametric time delay forms the basis of a connectivity pattern, which includes the temporal and time lagged dynamics of the synchronization between two sources. We model and discuss potential artifacts. We find that the general phase measures are remarkably stable. They produce highly comparable results for stochastic and periodic systems. Moreover, the methods proves useful for identifying brief periods of phase coupling and delays. Therefore, we propose that the method is useful as a basis for generating potential functional connective models. PMID- 21629587 TI - Synchronized changes to relative neuron populations in postnatal human neocortical development. AB - Mammalian prenatal neocortical development is dominated by the synchronized formation of the laminae and migration of neurons. Postnatal development likewise contains "sensitive periods" during which functions such as ocular dominance emerge. Here we introduce a novel neuroinformatics approach to identify and study these periods of active development. Although many aspects of the approach can be used in other studies, some specific techniques were chosen because of a legacy dataset of human histological data (Conel in The postnatal development of the human cerebral cortex, vol 1-8. Harvard University Press, Cambridge, 1939-1967). Our method calculates normalized change vectors from the raw histological data, and then employs k-means cluster analysis of the change vectors to explore the population dynamics of neurons from 37 neocortical areas across eight postnatal developmental stages from birth to 72 months in 54 subjects. We show that the cortical "address" (Brodmann area/sub-area and layer) provides the necessary resolution to segregate neuron population changes into seven correlated "k clusters" in k-means cluster analysis. The members in each k-cluster share a single change interval where the relative share of the cortex by the members undergoes its maximum change. The maximum change occurs in a different change interval for each k-cluster. Each k-cluster has at least one totally connected maximal "clique" which appears to correspond to cortical function. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11571 010-9103-3) contains supplementary material, which is available to authorized users. PMID- 21629588 TI - Mean square exponential and robust stability of stochastic discrete-time genetic regulatory networks with uncertainties. AB - This paper aims to analyze global robust exponential stability in the mean square sense of stochastic discrete-time genetic regulatory networks with stochastic delays and parameter uncertainties. Comparing to the previous research works, time-varying delays are assumed to be stochastic whose variation ranges and probability distributions of the time-varying delays are explored. Based on the stochastic analysis approach and some analysis techniques, several sufficient criteria for the global robust exponential stability in the mean square sense of the networks are derived. Moreover, two numerical examples are presented to show the effectiveness of the obtained results. PMID- 21629589 TI - Electronic pathway in reaction centers from Rhodobacter sphaeroides and Chloroflexus aurantiacus. AB - The reaction centers (RC) of Chloroflexus aurantiacus and Rhodobacter sphaeroidesH(M182)L mutant were investigated. Prediction for electron transfer (ET) at very low temperatures was also performed. To describe the kinetics of the C. aurantiacus RCs, the incoherent model of electron transfer was used. It was shown that the asymmetry in electronic coupling parameters must be included to explain the experiments. For the description of R. sphaeroidesH(M182)L mutant RCs, the coherent and incoherent models of electron transfer were used. These two models are discussed with regard to the observed electron transfer kinetics. It seems likely that the electron transfer asymmetry in R. sphaeroides RCs is caused mainly by the asymmetry in the free energy levels of L- and M-side cofactors. In the case of C. aurantiacus RCs, the unidirectionality of the charge separation can be caused mainly by the difference in the electronic coupling parameters in two branches. PMID- 21629590 TI - Mean-squared atomic displacements in hydrated lysozyme, native and denatured. AB - We use elastic neutron scattering to demonstrate that a sharp increase in the mean-squared atomic displacements, commonly observed in hydrated proteins above 200 K and often referred to as the dynamical transition, is present in the hydrated state of both native and denatured lysozyme. A direct comparison of the native and denatured protein thus confirms that the presence of the transition in the mean-squared atomic displacements is not specific to biologically functional molecules. PMID- 21629591 TI - Isoprenaline increases the slopes of restitution trajectory in the conscious rabbit with ischemic heart failure. AB - Roughly speaking, restitution is the dependence of recovery time of cardiac electrical activity on heart rate. Increased restitution slope is theorized to be predictive of sudden death after heart injury such as from coronary artery occlusion (ischemia). Adrenaline analogs are known to increase restitution slope in normal hearts, but their effects in failing hearts are unknown. Twenty-six rabbits underwent coronary ligation (n = 15) or sham surgery (n = 11) and implantation of a lead in the heart for recording electrocardiograms. Eight weeks later, unanesthetized rabbits were given 0.25-2.0 ml of 1 MUmol/L isoprenaline intravenously, which increased heart rate. Heart rate was quantified by time between QRS peaks (RR) and heart activity duration by R to T peak time (QTp). Ligated rabbits (n = 6) had lower ejection fraction than sham rabbits (n = 7, p < 0.0001) indicative of heart failure, but similar baseline RR (269 +/- 15 vs 292 +/- 23 ms, p = 0.07), QTp (104 +/- 17 vs 91 +/- 9 ms, p = 0.1), and isoprenaline induced minimum RR (204 +/- 11 vs 208 +/- 6 ms, p = 0.4). The trajectory of QTp vs TQ plots displayed hysteresis and regions of negative slope. The slope of the positive slope region was >1 in ligated rabbits (1.27 +/- 0.66) and <1 in sham rabbits (0.35 +/- 0.14, p = 0.004). The absolute value of the negative slope was greater in ligated rabbits (- 0.81 +/- 0.52 vs - 0.35 +/- 0.14, p = 0.04). Isoprenaline increased heart rate and slopes of restitution trajectory in failing hearts. The dynamics of restitution trajectory may hold clues for sudden death in heart failure patients. PMID- 21629592 TI - Bacterium in a box: sensing of quorum and environment by the LuxI/LuxR gene regulatory circuit. AB - The chemical signaling mechanism known as "bacterial quorum sensing" (QS) is normally interpreted as allowing bacteria to detect their own population density, in order to coordinate gene expression across a colony. However, the release of the chemical signal can also be interpreted as a means for one or a few cells to probe the local physical properties of their microenvironment. We have studied the behavior of the LuxI/LuxR QS circuit of Vibrio fischeri in tightly confining environments where individual cells detect their own released signals. We find that the lux genes become activated in these environments, although the activation onset time shows substantial cell-to-cell variability and little sensitivity to the confining volume. Our data suggest that noise in gene expression could significantly impact the utility of LuxI/LuxR as a probe of the local physical environment. PMID- 21629593 TI - The role of physical therapists in natural disasters: what can we learn from the earthquake in haiti? PMID- 21629594 TI - Patient Safety and Physiotherapy: What Does it Mean for Your Clinical Practice? PMID- 21629595 TI - Psychosocial treatment techniques to augment the impact of physiotherapy interventions for low back pain. AB - PURPOSE: The present study examined the profile of physical and psychosocial changes that occur in physiotherapy intervention when patients also participate in a psychosocial intervention. The psychosocial intervention, delivered by physiotherapists, was designed to target catastrophic thinking, fear of pain, perceived disability, and depression. METHODS: The study sample consisted of 48 individuals referred for the rehabilitation treatment of disabling back pain. Half the sample was enrolled in a physiotherapy intervention only; the other half was enrolled in a psychosocial intervention in addition to receiving a physiotherapy intervention. RESULTS: At post-treatment, the two treatment groups did not differ significantly on measures of pain severity, physical function, or self-reported disability. Patients who participated in the psychosocial intervention in addition to physiotherapy showed significantly greater reductions in pain catastrophizing, fear of movement, and depression than patients who received only the physiotherapy intervention. Reductions in psychosocial risk factors contributed to reduced use of the health care system, reduced use of pain medication, and improved return-to-work outcomes. CONCLUSIONS: The findings of the present study suggest that a psychosocial intervention provided by physiotherapists can lead to meaningful reductions in psychosocial risk factors for pain and disability and may contribute to more positive rehabilitation outcomes. PMID- 21629597 TI - Evidence-Based Management of an Individual Living with HIV. PMID- 21629596 TI - Neuropathic pain in patients with upper-extremity nerve injury. AB - PURPOSE: The purpose of this review was to present an analysis of the literature of the outcome studies reported in patients following traumatic upper-extremity (UE) nerve injuries (excluding amputation), to assess the presence of an association between neuropathic pain and outcome in patients following traumatic UE nerve injuries, and to provide recommendations for inclusion of more comprehensive outcome measures by clinicians who treat these patients. SUMMARY OF KEY POINTS: A Medline and CINAHL literature search retrieved 48 articles. This review identified very few studies of patients with peripheral nerve injury that reported neuropathic pain. When pain was reported, visual analogue or numeric rating scales were most frequently used; standardized questionnaires measuring pain or psychosocial function were rarely administered. Recent evidence shows substantial long-term disability and pain in patients following peripheral nerve injury. RECOMMENDATION: To better understand neuropathic pain in patients following peripheral nerve injury, future outcome studies should include valid, reliable measures of physical impairment, pain, disability, health-related quality of life, and psychosocial functioning. PMID- 21629598 TI - Educational needs of patients undergoing total joint arthroplasty. AB - PURPOSE: To identify the educational needs of adults who undergo total hip and total knee replacement surgery. METHODS: A qualitative research design using a semi-standardized interviewing method was employed. A purposive sampling technique was used to recruit participants, who were eligible if they were scheduled to undergo total hip or total knee replacement or had undergone total hip or total knee replacement in the previous 3 to 6 months. A comparative contrast method of analysis was used. RESULTS: Of 22 potential participants who were approached, 15 participated. Five were booked for upcoming total hip or total knee replacement and 10 had undergone at least one total hip or total knee replacement in the previous 3 to 6 months. Several themes related to specific educational needs and factors affecting educational needs, including access, preoperative phase, surgery and medical recovery, rehabilitation process and functional recovery, fears, and expectations counterbalanced with responsibility, emerged from the interviews. CONCLUSIONS: Educational needs of adults who undergo total hip and knee replacement surgery encompass a broad range of topics, confirming the importance of offering an all-inclusive information package regarding total hip and total knee replacement. PMID- 21629599 TI - Physical therapy management for adult patients undergoing cardiac surgery: a canadian practice survey. AB - PURPOSE: To determine current Canadian physical therapy practice for adult patients requiring routine care following cardiac surgery. METHODS: A telephone survey was conducted of a selected sample (n=18) of Canadian hospitals performing cardiac surgery to determine cardiorespiratory care, mobility, exercises, and education provided to patients undergoing cardiac surgery. RESULTS: An average of 21 cardiac surgeries per week (range: 6-42) were performed, with an average length of stay of 6.4 days (range: 4.0-10.6). Patients were seen preoperatively at 7 of 18 sites and on postoperative day 1 (POD-1) at 16 of 18 sites. On POD-1, 16 sites performed deep breathing and coughing, 7 used incentive spirometers, 13 did upper-extremity exercises, and 12 did lower-extremity exercises. Nine sites provided cardiorespiratory treatment on POD-3. On POD-1, patients were dangled at 17 sites and mobilized out of bed at 13. By POD-3, patients ambulated 50-120 m per session 2-5 times per day. Sternal precautions were variable, but the lifting limit was reported as ranging between 5 lb and 10 lb. CONCLUSIONS: Canadian physical therapists reported the provision of cardiorespiratory treatment after POD-1. According to current available evidence, this level of care may be unnecessary for uncomplicated patients following cardiac surgery. In addition, some sites provide cardiorespiratory treatment techniques that are not supported by evidence in the literature. Further research is required. PMID- 21629600 TI - Clinician's Commentary. PMID- 21629601 TI - Discharge from outpatient orthopaedic physiotherapy: a qualitative descriptive study of physiotherapists' practices. AB - PURPOSE: To describe the clinical and contextual factors that influence physiotherapists' discharge decision-making processes in outpatient orthopaedic settings. METHODS: The study used a descriptive qualitative design that included three key-informant interviews and two focus groups (n=7) of orthopaedic physiotherapists (total n=10) working in the Greater Toronto Area. Interviews and focus groups followed a semi-structured interview guide that included questions pertaining to participants' discharge decision-making processes, salient contextual factors, and challenges. Data were coded and analyzed for emerging categories and themes using constant comparison techniques and group analyses. RESULTS: Participants indicated that a combination of factors and strategies were brought to bear on discharge decision making but that the process changed with clinical experience. Over time, further emphasis was attributed to the patient's role in his or her rehabilitation, and self-management goals were increasingly promoted. Experience affected how therapists conceptualized their roles in discharge decisions and how they negotiated goals with patients. CONCLUSIONS: Discharge decision making is a complex process that requires integrating numerous factors and negotiating with patients. Physiotherapists' decision-making practices evolve with experience and reveal the complexity of implementing models of "client-centred care" in practice. Further research is needed to explore these findings in other settings. PMID- 21629602 TI - Effect of Anterior Tibiofemoral Glides on Knee Extension during Gait in Patients with Decreased Range of Motion after Anterior Cruciate Ligament Reconstruction. AB - PURPOSE: The purpose of this preliminary investigation was to evaluate the effect of anterior tibiofemoral glides on maximal knee extension and selected spatiotemporal characteristics during gait in patients with knee extension deficits after anterior cruciate ligament (ACL) reconstruction. METHODS: Twelve patients with knee-extension deficits after recent ACL reconstructions underwent quantitative gait analyses immediately before and after 10 minutes of repeated anterior tibiofemoral glides on the operative limb, and again after a 10-minute seated rest period. RESULTS: Maximum knee extension during stance phase of the operative limb significantly increased immediately after the treatment (mean increase: 2.0 degrees +/-4.1 degrees , 95% CI: 0.6 degrees -3.3 degrees ). Maximum knee extension decreased after the 10-minute rest period (mean decrease: 0.9 degrees +/-1.8 degrees , 95% CI: -0.1 degrees -1.8 degrees ), although the decrease was not statistically significant. Small increases in operative limb step length, stride length, and gait speed were observed after the rest period compared to baseline values only. CONCLUSIONS: A single session of anterior tibiofemoral glides increases maximal knee extension during the stance phase of gait in patients with knee-extension deficits. Increases in knee extension are small and short-lived, however, suggesting that continued activity is required to maintain the observed improvements. PMID- 21629603 TI - The effect of abdominal support on functional outcomes in patients following major abdominal surgery: a randomized controlled trial. AB - PURPOSE: Immobility and pain are modifiable risk factors for development of venous thromboembolism and pulmonary morbidity after major abdominal surgery (MAS). The purpose of this study was to investigate the effect of abdominal incision support with an elasticized abdominal binder on postoperative walk performance (mobility), perceived distress, pain, and pulmonary function in patients following MAS. METHODS: Seventy-five patients scheduled to undergo MAS via laparotomy were randomized to experimental (binder) or control (no binder) groups. Sixty (33 male, 27 female; mean age 58+/-14.9 years) completed the study. Preoperative measurements of 6-minute walk test (6MWT) distance, perceived distress, pain, and pulmonary function were repeated 1, 3, and 5 days after surgery. RESULTS: Surgery was associated with marked postoperative reductions (p<0.001) in walk distance (~75-78%, day 3) and forced vital capacity (35%, all days) for both groups. Improved 6MWT distance by day 5 was greater (p<0.05) for patients wearing a binder (80%) than for the control group (48%). Pain and symptom-associated distress remained unchanged following surgery with binder usage, increasing significantly (p<0.05) only in the no binder group. CONCLUSION: Elasticized abdominal binders provide a non-invasive intervention for enhancing recovery of walk performance, controlling pain and distress, and improving patients' experience following MAS. PMID- 21629604 TI - Cessation of exercise in the institutionalized elderly: effects on physical function. AB - PURPOSE: The objectives of this study were (1) to examine the effects of a 12 week exercise cessation period separating two 12-week exercise sessions on physical variables in an elderly institutionalized population and (2) to explore overall programme effectiveness. METHODS: The functional mobility of 25 elderly institutionalized adults participating in an existing exercise programme was examined using a one-group, interrupted time-series design. Functional mobility was evaluated at four time points during two cycles of a 12-week exercise programme, alternating with a 12-week period of no formal exercise. RESULTS: The primary outcome was the change in functional mobility scores, assessed at baseline, 12, 24 and 36 weeks. In both the higher-functioning (HF) group and the lower-functioning (LF) group, the cessation of exercise was associated with deterioration in physical function. The overall non-continuous nature of the programming under study also seemed to be detrimental to the physical function of the LF group. CONCLUSIONS: Findings of this small-sample study support the need to modify common practices in exercise programming for older, institutionalized people. PMID- 21629605 TI - Examining international clinical internships for canadian physical therapy students from 1997 to 2007. AB - PURPOSE: To describe international clinical internships (ICIs) for Canadian physical therapy (PT) students, explore the experiences of individuals involved in ICIs, and develop recommendations for future ICIs based on these findings. METHODS: This study employed a mixed-methods approach. An online questionnaire surveyed academic coordinators of clinical education (ACCEs, n=14) on the availability, destinations, and number of ICIs from 1997 to 2007. Semi-structured telephone interviews were then conducted with eight PT students, seven ACCEs, and three supervising clinicians to investigate their ICI experiences. Interview transcripts were coded descriptively and thematically using NVivo. RESULTS: ICIs are currently available at 12 of 14 Canadian PT schools. A total of 313 students participated in ICIs in 51 different destination countries from 1997 to 2007. Over this period, increasing numbers of students participated in ICIs and developing countries represented an increasing proportion of ICI destinations. Key themes identified in the interviews were opportunities, challenges, and facilitating factors. CONCLUSIONS: ICIs present unique opportunities for Canadian PT students. Recommendations to enhance the quality of future ICIs are (1) clearly defined objectives for ICIs, (2) additional follow-up post-ICI, and (3) improved record keeping and sharing of information on ICI destination countries and host sites. PMID- 21629606 TI - Clinician's Commentary. PMID- 21629607 TI - Hereditary hemochromatosis: a literature review and case report. AB - PURPOSE: To improve understanding in the physical therapy (PT) community of hereditary hemochromatosis (HH), a common but little-known iron overload disorder, symptoms of which may mimic other orthopaedic conditions. Medical management typically involves phlebotomy to remove excess iron; however, there is little specific information in the literature on PT management of patients with HH after trauma. CASE DESCRIPTION: The patient was a 65-year-old woman with multiple fall-related traumas, including right wrist, thumb, and patellar fractures and left thigh muscle strain with significant ecchymosis and effusion. Medical history included HH. Iron-related lab values had been analyzed 9 days prior to the fall and had demonstrated a steady increase over the previous 4 months since her last phlebotomy. OUTCOMES: As the level of exercise and activity increased during the course of PT treatment, the patient developed shortness of breath and increased fatigue. The exercise level in therapy was reduced to accommodate the change in the patient's response. Blood values analyzed 7 weeks after the fall demonstrated a drop in haemoglobin and hematocrit values, while serum ferritin levels had risen. IMPLICATIONS: Understanding early symptoms and management of a patient with manifestations of HH will better enable physical therapists to consider this disorder as a differential diagnosis or co-morbidity that affects treatment considerations. PMID- 21629608 TI - What Does the Cochrane Collaboration Say about Therapeutic Ultrasound? PMID- 21629609 TI - Systemic therapy of New World cutaneous leishmaniasis: A case report and review article. AB - Cutaneous leishmaniasis is a disease endemic to Central and South America, Mexico and the Caribbean, and affects millions of people. As travel to these regions becomes more common, cutaneous leishmaniasis is becoming a disease of increasing importance in the developed world. However, disease recognition and access to appropriate therapy for cutaneous leishmaniasis remains a challenge in North America. The present article reports a case of cutaneous leishmaniasis in a Canadian man following a trip to Costa Rica. Species-specific diagnosis was confirmed by polymerase chain reaction analysis of a skin biopsy, which was positive for Leishmania panamensis. After failing a course of itraconazole, the patient was successfully treated with sodium stibogluconate, despite significant barriers to administering this therapy, and the paucity of data regarding its efficacy and tolerability. The pathophysiology, diagnosis and systemic treatment of cutaneous leishmaniasis, as well as its emerging presence in the developed world, are reviewed. PMID- 21629610 TI - Risk of coinfection with Chlamydia trachomatis and Neisseria gonorrhoeae in Nova Scotia. AB - BACKGROUND: The frequency of Chlamydia trachomatis and Neisseria gonorrhoeae coinfection can vary depending on their individual incidence and prevalence rates. OBJECTIVE: To determine the frequency of C trachomatis and N gonorrhoeae coinfections by evaluating the results of testing in 2007 and 2008 to better inform testing and treatment decisions. METHODS: Specimens from the same patient submitted on the same day served as the basis for the present study. The age, sex and the source of the specimen were also linked to the accession number. Infection and coinfection rates were analyzed in both males and females. RESULTS: Concurrent testing was performed on 41,567 female specimens and 1827 male specimens, of which, 1495 female samples (3.6%) tested positive for C trachomatis infection and 88 (0.2%) tested positive for N gonorrhoeae infections. Only 31 females were coinfected; however, for those between 11 and 25 years of age, 25 of 61 females (40.1%) with N gonorrhoeae infection also tested positive for C trachomatis infection; conversely, 25 of 1248 females (2.0%) with C trachomatis infection also tested positive for N gonorrhoeae infection. For males, 213 (11.7%) tested positive for C trachomatis infection, and 59 (3.2%) tested positive for N gonorrhoeae infection. In 30 males with N gonorrhoeae between 11 and 25 years of age, and 149 males with C trachomatis, eight coinfections were observed (26.7% and 5.3%, respectively). Of those older than 25 years of age, only five of 905 men and six of 19,465 women were coinfected. None of the 10,935 women who were 30 years of age or older had coinfections. CONCLUSION: The N gonorrhoeae coinfection rate in males with C trachomatis may justify empirical antimicrobials; however, in females, the proportion of coinfected may not justify empirical treatment for N gonorrhoeae infection when the C trachomatis test is positive and N gonorrhoeae testing has not been performed. PMID- 21629611 TI - Adamantane resistance in seasonal human influenza A viruses from Calgary, Alberta (January 2007 to August 2008). AB - The available antivirals for the treatment and prophylaxis of influenza A infections include the adamantanes (amantadine and rimantadine), which are matrix (M2) protein inhibitors, and the neuraminidase inhibitors (oseltamivir and zanamivir). Resistance to the adamantanes is conferred by mutations at amino acid positions 26, 27, 30, 31 or 34 within the M2 protein of influenza A viruses. A significant increase in adamantane resistance has been reported worldwide since 2003, reflected by a similar increase in Canada. The present study reports on the frequency of adamantane resistance in seasonal influenza A viruses in Calgary, Alberta, for the period between January 2007 and August 2008, as an update to the previous report. Positive influenza A samples (221 original patient specimens and 34 isolates obtained by viral culture) were analyzed for changes in the critical amino acid residues of the M2 gene. The amplification and sequencing of regions that confer adamantane resistance directly from RNA extracts of clinical samples (without previous culture) makes this approach a fast and efficient process for monitoring resistance. The results showed that the frequency of resistance varied from 37.5% to 49.2% in circulating influenza A H3N2 virus strains (n=213) between January 2007 and April 2007. The frequency of resistance increased to 100% in May 2007, after which all H3N2 viruses were resistant until the end of the monitoring period. All resistant H3N2 viruses contained the serine to asparagine substitution at amino acid position 31. Resistance was not observed in the H1N1 viruses tested (n=39) within this monitoring period. The level of adamantane resistance in H3N2 viruses continues to remain high since resistant viruses became the prevalent circulating strains in 2005. Recent reports have indicated that the currently circulating swine-origin influenza A H1N1 subtype viruses are adamantane resistant. It is, thus, important to continue to monitor seasonal influenza A viruses for antiviral resistance markers to ensure optimal prophylaxis and treatment. PMID- 21629612 TI - A descriptive review of selected nonviral enteric illnesses reported in children in Quebec between 1999 and 2006. AB - OBJECTIVE: To review the epidemiology of selected nonviral enteric illnesses reported in children in Quebec between 1999 and 2006. METHODS: Incidence rates were calculated to describe age, sex, temporal and geographical characteristics of the selected nonviral enteric cases reported in children who were between zero and four years of age. Standard descriptive methods were used to analyze the temporal and geographical distributions of the incidence rates. RESULTS: A total of 5068 cases were reported. Of these, three pathogens accounted for the majority of the infections: Giardia (32.52%), Salmonella (30.98%) and Campylobacter (30.82%). Salmonella was most frequent in children younger than one year of age, whereas comparable incidence rates for the three pathogens were calculated for children between one and four years of age. For Giardia, the geographical distributions showed that the highest rates were in areas with more than 100,000 inhabitants (except Montreal, Quebec); for Salmonella, the highest rates were in Montreal; and for Campylobacter, the highest rates were in areas with fewer than 10,000 inhabitants. No detectable trends were seen over the study period for the three pathogens. Seasonal summer peaks were noted for Salmonella and Campylobacter, contrasting with late summer to early autumn peaks for Giardia. CONCLUSION: Findings suggest that Giardia, Salmonella and Campylobacter were the most common causes of nonviral enteric illnesses reported in children in Quebec. Giardia cases seemed to arise from different sources and transmission routes than the other two pathogens. Characteristics specific to Campylobacter infections in children, namely its predominance in areas with low population densities, and to Salmonella infections, namely predominance in the Greater Montreal area, should be further investigated to better guide prevention and control measures. PMID- 21629613 TI - New antibacterials? PMID- 21629614 TI - Infective endocarditis: Updated guidelines. AB - The most recent revision of the American Heart Association guidelines on infective endocarditis prophylaxis occurred in 2007. These revisions were based on the fact that current data have brought into question the benefit of previous recommendations for infective endocarditis prophylaxis. It was noted that the bacteremia that occurs following dental procedures represents only a fraction of the episodes of bacteremia that occur with activities of daily living (such as chewing, brushing teeth and other oral hygiene measures). The target groups and the procedures for which prophylaxis is reasonable have been significantly reduced in number. The focus is now on patients who are most likely to have adverse outcomes from infectious endocarditis. The present article is targeted at practicing Canadian physicians and provides the rationale for the current recommendations. In addition to a summary of the indications for prophylaxis, information is provided on the conditions for which prophylaxis is not recommended. PMID- 21629615 TI - A 10-year retrospective review of Salmonella infections at the Children's Hospital in London, Ontario. AB - OBJECTIVES: To describe Salmonella infections in children presenting to the Children's Hospital (London Health Sciences Centre, London, Ontario), to assess risk factors for infection and to examine whether younger children, particularly infants younger than 12 weeks of age, experience higher morbidity than older children. METHODS: A 10-year retrospective review of children with Salmonella infections at the Children's Hospital was conducted. Patient demographics, risk factors for infection, clinical characteristics, bacteriology and outcome were collected from the hospital charts and laboratory records. Data were separated into groups based on age and recent use of antibiotics to analyze differences in outcomes. RESULTS: Sixty-six children with Salmonella infections presented to the Children's Hospital over a 10-year period. Common risk factors for Salmonella infection included having sick contacts, living in a rural area, recent travel, contact with pets (especially reptiles) and exposure to local water. Younger age was associated with an increased likelihood of admission to hospital, treatment with antibiotics and a longer course of antibiotic therapy. This was true when comparing older infants with those younger than 12 weeks of age. Patients recently treated with antibiotics and those with significant underlying medical conditions were more likely to be admitted. CONCLUSIONS: A wider knowledge of the epidemiological risk factors for Salmonella infection may improve diagnosis. Higher admission rates were expected in children younger than 12 weeks of age, those recently treated with antibiotics and those who had a significant underlying medical condition. A prospective, multicentre study is needed to further address questions regarding increased illness severity and appropriate management of Salmonella infections in children younger than 12 weeks of age. PMID- 21629617 TI - Methicillin-resistant Staphylococcus aureus endocarditis and de novo development of daptomycin resistance during therapy. AB - Daptomycin resistance in Staphylococcus aureus has been previously reported, but the development of resistance while on therapy with subsequent clinical failure for endocarditis has been infrequently reported. A case of persistent methicillin resistant S aureus (MRSA) bacteremia in the setting of right-sided endocarditis in a 38-year-old man with a history of intravenous drug use is presented. He developed de novo resistance to daptomycin during therapy after several courses of antibiotics, with subsequent clinical failure. Isolates were identified by molecular characterization to be community-acquired MRSA 10 (USA300). To the authors' knowledge, the present case was the first in Canada to involve the de novo development of daptomycin resistance with clinical failure due to MRSA during therapy for endocarditis. Clinicians and microbiologists must be aware of this phenomenon given the implications for treatment and transmission of the strain. It also raises questions regarding the use of daptomycin in settings of heavily pretreated patients with persistent MRSA bacteremia. PMID- 21629616 TI - Systematic review of invasive Acinetobacter infections in children. AB - INTRODUCTION: Clinicians are generally familiar with Acinetobacter as an etiological agent for serious nosocomial infections in intensive care units. However, there are no previous reviews of the full spectrum of invasive infections in children. METHODS: A systematic review of the literature was completed up to December 2008 for reports of invasive Acinetobacter infections in children. RESULTS: There were 101 studies that met the inclusion criteria including 18 possible outbreaks, 33 case series and 49 case reports. Suspected outbreaks were concentrated in neonatal intensive care units (16 of 18 outbreaks) and involved bacteremia or meningitis. Proof of isolate clonality or identification of the source of the outbreak was seldom established. Case series were primarily of children younger than five years of age presenting with bacteremia (sometimes multiresistant), meningitis, endocarditis or endophthalmitis, with many community-acquired infections being reported from India. Case reports consisted of unique presentations of disease or the use of novel therapies. Attributable mortality in the outbreaks and case series combined was 68 of 469 (14.5%). DISCUSSION: Invasive Acinetobacter infections in children usually manifest as bacteremia, meningitis or both, but can result in a wide variety of clinical presentations. Outbreaks are primarily a problem in newborns with underlying medical conditions. Most reports of community-acquired infections are from tropical countries. The study of the mechanism of colonization and infection of children in intensive care units and of neonates in tropical countries may provide some insight into prevention of invasive infections. PMID- 21629618 TI - A new method in the treatment of postburn and post-traumatic scar contractures: Double-opposing Z- and V- (K-M-N) plasty. AB - BACKGROUND: To date, many techniques for the surgical treatment of postburn scar contractures have been described. Some of the most popular techniques are Z plasty, V-Y-plasty and their analogues. A major limitation of these techniques is that the excess tissue requires excision of the dog ear. The current study presents a new modification of the double-opposing Z- and V-plasty, called 'K-M-N plasty'. METHODS: Twenty postburn scar contractures were successfully treated with K-M-N plasty. The postoperative results depict the versatility of this technique in the surgical treatment of postburn scar contractures, especially in the upper and lower extremities. RESULTS: There was no distal flap necrosis, and postoperative recovery was uneventful in all operated patients. K-M-N plasty is an effective and alternative method for the surgical treatment of postburn scar contractures. In addition, drawing and flap transpositions were not complicated. DISCUSSION: THERE ARE MANY ADVANTAGES TO USING THIS TECHNIQUE: K-M-N plasty can be safely used when skin tension crosses the contracture line; it is superior to other local flaps because of its rich vascularity and mobility for superficial scars; it can be recommended to the inexperienced surgeon because it can be performed with ease; it is also an effective procedure for the pericontracture area due to its V limb (it can prevent recontracture); the colour and texture matches are more cosmetically acceptable, and the resultant contracture release is similar to other techniques; the dog ear formation is not seen; it can be performed under local anesthesia in most cases (not in children); and it has a shorter period of operation and hospitalization than other techniques. PMID- 21629619 TI - Acute symptomatic hematoma with defined etiology seven years after breast reconstruction: A case report and literature review. AB - Augmentation mammaplasty with implants can be complicated by hematoma formation. The majority of hematomas occur in the immediate postoperative period. There are, however, some reports of hematomas with a delayed presentation. These hematomas in the late postoperative period are rare, and many of these cases do not have a definitive mechanism of injury or develop symptoms immediately after the triggering event. A case of late capsular hematoma seven years after breast reconstruction with a saline implant following mastectomy is presented. In contrast to the majority of published cases, the patient presented with both a known and traceable cause of her bleed and with symptoms that developed immediately after the initial event. Furthermore, the mechanism of her injury was consistent with a bleeding capsular tear that was observed intraoperatively. The present case emphasizes the importance of thorough evaluation and management of patients with a history of breast reconstruction. PMID- 21629620 TI - Mucormycosis infection following intravenous access in the forearm. AB - Mucormycosis is an opportunistic infection that is often fatal, requiring aggressive local control as well as systemic therapy. A rare case of a forearm infection originating in a traumatic intravenous access portal is described in the present study. The Mucor species infection prevented liver transplant, and the patient passed away. In the present case, it was decided to limit the resection to the skin and subcutaneous tissue based on a frozen section and the viability of the biopsied tissue. With consistently rising numbers of immunocompromised patients, awareness and familiarity with mucormycosis in the extremities is important. Knowing that a minimal traumatic event may precede the infection could assist in prevention and early diagnosis. Guidelines for pathological and clinical diagnosis and treatment need to be further clarified. PMID- 21629621 TI - David e. Beck, m.d. PMID- 21629622 TI - Urology and gynecology. PMID- 21629623 TI - Adnexal masses and malignancies of importance to the colorectal surgeon. AB - In this article, the authors review both benign and malignant ovarian masses, as the colorectal surgeon who encounters an adnexal mass at the time of surgery should be aware of the steps necessary for surgical staging and optimal tumor resection.Ovarian tumors-most of which are benign-are divided into three major categories, in order of frequency: epithelial, germ cell, and sex cord-stromal tumors. Nonneoplastic conditions of the ovary that may present as adnexal masses include the following, according to World Health Organization (WHO) classification: pregnancy luteoma, hyperplasia of ovarian stroma, hyperthecosis, massive edema, solitary follicle cysts and corpus luteal cysts, multiple follicle cysts, and endometriosis.Epithelial ovarian tumors arise from the surface epithelium and can be benign or malignant. Histologic types are serous, mucinous, endometrioid, clear cell, or Brenner. Germ cell tumors are more likely to appear in females under 20 years, accounting for 70% of ovarian tumors in this age group. Approximately 3% are malignant. Teratomas are the most common germ cell tumors. Malignancies, in addition to malignant teratomas, include dysgerminomas, endodermal sinus tumors, and embryonal carcinomas. The more common sex cord stromal tumors include granulosa stromal cell tumors, Sertoli-Leydig cell tumors, and gynandroblastomas.Surgical staging and optimal tumor resection are also addressed, with a focus on epithelial malignancies, as they are the most relevant to colorectal surgeons. PMID- 21629624 TI - Endometriosis for the colorectal surgeon. AB - Endometriosis is defined as the presence of endometrial glands and stroma outside the uterine cavity. Although the exact pathophysiology is unclear, endometriosis is a well-known cause of pelvic pain and infertility in reproductive-aged women. Endometriosis can have extrapelvic manifestations relevant for colorectal surgeons to appreciate, such as cyclic constipation, diarrhea, hematochezia, and dyschezia. The treatment of endometriosis involves a combination of medical and surgical interventions where close collaboration between the gynecologist and colorectal surgeon can help achieve prolonged periods of symptom remission. PMID- 21629626 TI - Rectocele. AB - Rectoceles are common and involve a herniation of the rectum into the posterior vaginal wall that results in a vaginal bulge. Women with rectoceles generally complain of perineal and vaginal pressure, obstructive defecation, constipation, or the need to splint or digitally reduce the vagina to effectuate a bowel movement. Rectoceles are associated with age and parturition and arise from either a tear or stretching of the rectovaginal fascia, and can be repaired via a vaginal, anal, or perineal approach. Although the rate of successful anatomic repair is high, reports of functional outcome are more variable. PMID- 21629625 TI - Gastrointestinal Conditions during Pregnancy. AB - Pregnancy causes anatomic and physiologic changes in the gastrointestinal tract. Pregnant women with intestinal disease such as Crohn disease or ulcerative colitis pose a management challenge in clinical diagnosis, radiologic evaluation, and treatment secondary to potential risk to the fetus. Heightened physician awareness on possible etiologies such as appendicitis, diverticulitis, and rarely colorectal cancer is required for rapid diagnosis and treatment to improve maternal/fetal outcome. A multidisciplinary approach to evaluation is a necessity because radiologic procedures and treatment medications commonly used in nonpregnant patients may have a potential harmful effect on the fetus. The authors review several gastrointestinal conditions encountered during pregnancy and address presentation, diagnosis, and treatment of each condition. PMID- 21629627 TI - Rectovaginal fistulas. AB - Rectovaginal fistulas are abnormal epithelial-lined connections between the rectum and vagina. They can be quite bothersome to both the patient and the surgeon due to their irritating and embarrassing symptoms and high failure rate after repair. An individualized, systematic approach to these fistulas based on their size, location, and etiology provides a more concise treatment plan. Treatment options of medical therapy, advancement flaps, plugs, fistula ligation, and tissue interposition are discussed. PMID- 21629628 TI - Urinary tract injures: recognition and management. AB - Iatrogenic injury to the urinary tract during colorectal surgery can be a source of significant morbidity. Although most cases of ureteral injury occur in patients without significant risk factors, the incidence of urinary tract injuries increases in patients with prior pelvic operations, inflammatory bowel disease, infection, and in patients with extensive neoplasms causing distortion of normal surgical planes. The most commonly injured locations are the ureter, bladder, and urethra. Mechanisms of injury include ligation, transection, devascularization, and energy induced. Early identification of urinary tract injuries is paramount in minimizing morbidity and preservation of renal function. Anatomic considerations for preventing injuries, diagnostic techniques for localizing and staging injuries, as well as reconstructive techniques and principles of repair are discussed. PMID- 21629629 TI - Bladder Reconstruction and Diversion during Colorectal Surgery. AB - Extirpative procedures for advanced colorectal cancers can involve multivisceral pelvic resections. In patients without evidence of distant metastatic disease, a reasonable therapeutic effect can be expected when negative surgical margins are obtained. For patients with bladder involvement, the decision to perform a bladder-sparing procedure or a total pelvic exenteration will be based on the extent of the primary lesion as well as patient characteristics. In this article, the authors describe bladder-sparing techniques with and without enterocystoplasty as well as options for urinary diversion in patients requiring total pelvic exenteration. Contraindications and clinical decision making regarding bladder reconstruction or replacement are discussed. PMID- 21629630 TI - Voiding dysfunction after pelvic colorectal surgery. AB - Bladder dysfunction following colorectal surgery may be related to extirpative procedures in the region of the pelvic autonomic plexus. The most common etiology is from autonomic disruption during abdominoperineal or low anterior resections. Contemporary technical modifications have allowed surgeons to achieve oncologic control while preserving the autonomic nerves that innervate the bladder and sexual organs. Although these modifications have resulted in a significant decrease in the incidence of postoperative bladder dysfunction, bladder dysfunction continues to be a source of significant morbidity after surgery. In this patient population, symptoms are not reliable for accurate diagnosis. The use of urodynamics provides objective measurements of bladder and outlet function and are paramount in providing an accurate diagnosis and in recommending treatments. Follow-up and treatment are highly individualized based on urodynamic findings, patient expectations, patient abilities, and family support. This article provides an overview of pertinent neuroanatomy, diagnosis, urodynamic interpretation, and treatment related to bladder dysfunction following pelvic colorectal surgery. PMID- 21629631 TI - On "pathogenesis and management of postoperative ileus" (clin colon rectal surg 2009;22:47-50). PMID- 21629632 TI - From Editor's Desk.... PMID- 21629634 TI - Comparative evaluation of simultaneous bone marrow aspiration and bone marrow biopsy: an institutional experience. AB - Bone marrow aspirations and bone marrow biopsies are important diagnostic procedures. A comparative study of both the procedures done simultaneously was retrospectively reviewed in 160 cases where the clinical history is correlated with BMA and BMB results. The advantage of each method is analyzed. Correlation of our findings with that given in the literature is done to give a guideline for both techniques. We have found that 61.25% of the cases showed a positive correlation between bone marrow aspiration and bone marrow biopsy. However, we found that tuberculous granulomas and Hodgkin disease involvement of the marrow were detected better in bone marrow biopsies. The advantage of both the procedures done together provided more material and enabled us to study the cytomorphology of the cells, with the pattern of distribution of the cells depending on the cases. However, when both the procedures are done simultaneously, a proper technique is required so as to yield good diagnostic material. PMID- 21629633 TI - Advances in haematological pharmacotherapy in 21st century. AB - Last quarter of twentieth century and the first 10 years of 21st century has seen phenomenal development in haematological pharmacotherapy. Tailor made chemotherapeutic agents, vast array of monoclonal antibodies, epigenetic modifiers, growth factors for red cells white cells and platelets, peptidomimetics as growth factors, newer thrombin inhibitors, safer plasma derived protein molecules, recombinant molecules, newer immunomodulators, enzyme replacement therapy and above all a plethora of targetted molecules targeting innumerable pathways involved in cell division, growth, proliferation and apoptosis has given immense number of clinically usable molecules in the hand of modern haematologists to treat diverse hitherto untreatable haematological disorders effectively. In addition many old molecules are finding newer uses in diverse fields, thalidomide as an antiangiogenic molecule is a prime example of this genre. Present overview has tried to capture this rapidly evolving area in a broad canvas without going into details of indications and contraindications of the use of various drugs. PMID- 21629635 TI - Serum immunoglobulins in patients with iron deficiency anemia. AB - Iron deficiency is a common cause of anemia worldwide. Documentation shows that these patients have impaired immunity and are prone to infections. The aim of this study was to confirm whether serum immunoglobulins change in adult nonpregnant females with iron deficiency anemia. Based on patients' clinical history, CBC results, and serum ferritin, iron and total iron binding capacity, 45 healthy patients and 45 iron deficiency anemia (hypochrom microcytic) patients were entered into this case-control study. Serum IgG, IgA and IgM were measured in patient and control groups using the nephelometry method. Both of the groups were compared using suitable statistical test. Mean serum values of IgG, IgA and IgM in the patient group were 12.47 +/- 7.67, 1.93 +/- 0.92 and 1.35 +/- 0.90 g/l respectively. Mean serum values for the control group were 12.51 +/- 6.85, 2.14 +/- 0.88 and 1.49 +/- 0.73 g/l respectively. Although serum immunoglobulins were slightly lower in the patient group compared to the control group these differences were not significant with the t test. Our study showed serum immunoglobulin levels did not change in iron deficiency anemia. PMID- 21629636 TI - Microbial etiology of febrile neutropenia. AB - Bacterial and fungal infections are a major cause of morbidity and mortality among neutropenic patients. The choice of empiric antimicrobial regimen is based on susceptibility pattern of locally prevalent pathogens. From 64 febrile neutropenic patients with clinical sepsis, blood and other appropriate clinical specimens were processed to determine bacterial and fungal spectrum and their antimicrobial susceptibility pattern. Risk factors for developing sepsis were determined by case-control study. 68 organisms were recovered. Fifteen (22.05%) were Gram-positive cocci with predominance of methicillin Sensitive S. aureus (10.29%), 47 (69.11%) were Gram-negative rods with predominance of Klebsiella pneumoniae (30.88%) and four were Non albicans Candida. 81% and 60% of Klebsiella and E. coli were ESBL producers. All species of Candida were sensitive to amphoterecin B and voriconazole. Duration and extent of neutropenia, chemotherapy, immunosuppressive therapy, altered mucosal barriers and presence of central venous lines were statistically significant risk factors for developing sepsis. Gram-negative bacteria were the predominant isolates. The choice of therapy in neutropenic patients should be formulated based on local spectrum of microbes and local and regional resistance patterns. PMID- 21629637 TI - Evaluation of Cytopenias Occurring in Imatinib Treated Chronic Myeloid Leukemia (CML) Patients. AB - Imatinib Mesylate, a Tyrosine Kinase inhibitor, is presently the drug of choice for Chronic myeloid leukemia (CML). During therapy, a few patients develop myelosuppression and present with cytopenias. To study the bone marrow morphology in imatinib treated CML patients presenting with persistent cytopenias. The cases were retrieved from the Hematopathology record files, Department of Pathology; the study period being January 2008-June 2009. Cases of CML on Imatinib presenting with grade 2 or more anemia, neutropenia and/or thrombocytopenias with bone marrow studies, were included in the study. The morphology of all cases was reviewed with cytogenetic studies. Follow-up details were obtained from the Medical Oncology records. During the study period, 683 Imatinib treated CML patients had bone marrow studies as part of their follow-up investigations. Of these, 60 patients (9%) had some form of persistent cytopenia. The patients ranged from 21 to 75 years of age with a median age of 38 years. The male:female ratio was 1:1. There were 46 patients with >=grade 2 anemia, 25 patients with >=grade 2 neutropenia and 37 patients with >=grade 2 thrombocytopenia. Of these, 18 patients had bicytopenia and 13 cases had pancytopenia. The marrow evaluation revealed morphologic response in 30 patients, persistent marrow disease in five patients, marrow hypoplasia in six patients, extensive stromal changes including fibrosis in five patients, megaloblastic erythropoiesis in 11 patients and disease progression to accelerated or blast crisis in three patients. Various degrees of cytopenias may occur in few patients of CML on imatinib therapy. Regular hematologic follow-up is required so that the drug may be stopped or dose modified as per the individual's needs. PMID- 21629638 TI - Unilateral massive pleural effusion: a presentation of unsuspected multiple myeloma. AB - Pleural effusion in case of multiple myeloma is usually multifactorial but effusion due to myelomatous deposition of pleura is extremely uncommon. We are reporting a unique case of unsuspected multiple myeloma presenting as left sided massive pleural effusion due to myelomatous deposit in pleura and a rare M-band in the alpha-2 region in serum protein electrophoresis. A 61 year lady presented with cough, weakness and progressive shortness of breath. Examination revealed mild pallor and left sided massive pleural effusion that re-accumulated despite treatment. PAP stained smear of pleural fluid showed a large number of plasma cells and pleural biopsy revealed infiltration of plasma cells. Fiber-optic bronchoscopy was inconclusive. Blood examination revealed high value of alpha-2 globulin. Serum electrophoresis showed M band in alpha-2 region and urine electrophoresis showed faint monoclonal light chain pattern. X-ray skull showed multiple punched out osteolytic lesions. Bone marrow examination revealed hypercellular marrow with atypical plasma cells including binucleate forms in large number (above 55% of nucleated cell population). PMID- 21629639 TI - Chronic disseminated histoplasmosis bone marrow involvement in an immunocompetent patient. AB - Histoplasmosis normally do not affect immunocompetent individuals. Prolonged exposure may cause chronic disseminated histoplasmosis. Elderly male patient presented with fever, hematuria and pain in right hypochondrium. He had pallor, fever and mild hepatosplenomegaly. Investigations revealed anemia and thrombocytopenia. Giemsa stained bone marrow aspirate showed yeast-like cells, suggestive of Histoplasma capsulatum. PAS stained bone marrow aspirate and biopsy confirmed the diagnosis. PMID- 21629640 TI - Factor VII Deficiency: A Rare Case Report. AB - Factor VII deficiency is a rare congenital coagulopathy. Prolonged prothrombin time with normal partial thromboplastin time indicates factor VII deficiency. For the definitive diagnosis, the specific factor VII level should be investigated. We report a 20 day old, baby girl, born full-term who was admitted with the diagnosis of sepsis. Hematological tests revealed prolonged prothrombin time and a factor VII level of nine percent. After antibiotic therapy and fresh frozen plasma replacement, her clinical status improved but the prothrombin time continued to be prolonged. On the seventh day of discharge the baby died due to sudden intracranial hemorrhage. PMID- 21629641 TI - Chest Wall Swelling, A Rare Presentation of Richter's Transformation. AB - A large cell lymphoma develops in approximately 1-10% of chronic lymphocytic leukemia (CLL). This is known as Richter's syndrome. Chest wall swelling is a very unusual presentation of this syndrome. Here we report another such case of chest wall swelling, a very rare presentation of Richter's syndrome. PMID- 21629642 TI - Melamine and its effect on coagulation system. PMID- 21629643 TI - Hemophagocytic lymphohistiocytosis in B-cell lymphoproliferative disorder: report of a rare association. PMID- 21629644 TI - Primary phacoemulsification and intraocular lens implantation for acute primary angle-closure. AB - BACKGROUND: To investigate the effect of primary phacoemulsification on intraocular pressure (IOP) in patients with acute primary angle-closure (PAC) and coexisting cataract. METHODOLOGY: Sixteen eyes of 14 patients with acute PAC received phacoemulsification and intraocular lens implantation as initial management for medically uncontrolled IOP in a retrospective chart review. The effects on IOP, vision, anterior chamber depth (ACD), and number of antiglaucoma medications were evaluated. PRINCIPAL FINDINGS: The postoperative IOP was reduced in 16 eyes (100%). The mean +/- standard deviation preoperative IOP was 48.81 +/- 16.83 mm Hg, which decreased postoperatively to 16.46 +/- 10.67 mm Hg at 1 day, 9.43 +/- 3.03 mm Hg at 1 week, 9.49 +/- 2.14 mm Hg at 2 weeks, 10.78 +/- 3.56 mm Hg at 1 month, and 10.70 +/- 2.80 mm Hg at 3 months (p < 0.001). The mean number of antiglaucoma medications decreased from 3.56 +/- 1.14 to 0.13 +/- 0.34 (p < 0.001). The average preoperative ACD was 2.08 +/- 0.35 mm, which increased to 3.59 +/- 0.33 mm after surgery (p < 0.001). Visual acuity (converted into logarithm of the minimum angle of resolution [logMAR]) improved from 1.14 +/- 0.71 to 0.73 +/- 0.53 (p = 0.001). CONCLUSIONS: Primary phacoemulsification plus intraocular lens implantation lowered IOP, reduced the use of antiglaucoma medications, and improved vision in patients with acute PAC. This is a safe and effective method of IOP control and can be considered a first treatment option in managing patients with acute PAC and coexisting cataract. PMID- 21629645 TI - A simple threshold rule is sufficient to explain sophisticated collective decision-making. AB - Decision-making animals can use slow-but-accurate strategies, such as making multiple comparisons, or opt for simpler, faster strategies to find a 'good enough' option. Social animals make collective decisions about many group behaviours including foraging and migration. The key to the collective choice lies with individual behaviour. We present a case study of a collective decision making process (house-hunting ants, Temnothorax albipennis), in which a previously proposed decision strategy involved both quality-dependent hesitancy and direct comparisons of nests by scouts. An alternative possible decision strategy is that scouting ants use a very simple quality-dependent threshold rule to decide whether to recruit nest-mates to a new site or search for alternatives. We use analytical and simulation modelling to demonstrate that this simple rule is sufficient to explain empirical patterns from three studies of collective decision-making in ants, and can account parsimoniously for apparent comparison by individuals and apparent hesitancy (recruitment latency) effects, when available nests differ strongly in quality. This highlights the need to carefully design experiments to detect individual comparison. We present empirical data strongly suggesting that best-of-n comparison is not used by individual ants, although individual sequential comparisons are not ruled out. However, by using a simple threshold rule, decision-making groups are able to effectively compare options, without relying on any form of direct comparison of alternatives by individuals. This parsimonious mechanism could promote collective rationality in group decision-making. PMID- 21629646 TI - Crystal structure analysis reveals functional flexibility in the selenocysteine specific tRNA from mouse. AB - BACKGROUND: Selenocysteine tRNAs (tRNA(Sec)) exhibit a number of unique identity elements that are recognized specifically by proteins of the selenocysteine biosynthetic pathways and decoding machineries. Presently, these identity elements and the mechanisms by which they are interpreted by tRNA(Sec) interacting factors are incompletely understood. METHODOLOGY/PRINCIPAL FINDINGS: We applied rational mutagenesis to obtain well diffracting crystals of murine tRNA(Sec). tRNA(Sec) lacking the single-stranded 3'-acceptor end ((DeltaGCCA)RNA(Sec)) yielded a crystal structure at 2.0 A resolution. The global structure of (DeltaGCCA)RNA(Sec) resembles the structure of human tRNA(Sec) determined at 3.1 A resolution. Structural comparisons revealed flexible regions in tRNA(Sec) used for induced fit binding to selenophosphate synthetase. Water molecules located in the present structure were involved in the stabilization of two alternative conformations of the anticodon stem-loop. Modeling of a 2'-O methylated ribose at position U34 of the anticodon loop as found in a sub population of tRNA(Sec)in vivo showed how this modification favors an anticodon loop conformation that is functional during decoding on the ribosome. Soaking of crystals in Mn(2+)-containing buffer revealed eight potential divalent metal ion binding sites but the located metal ions did not significantly stabilize specific structural features of tRNA(Sec). CONCLUSIONS/SIGNIFICANCE: We provide the most highly resolved structure of a tRNA(Sec) molecule to date and assessed the influence of water molecules and metal ions on the molecule's conformation and dynamics. Our results suggest how conformational changes of tRNA(Sec) support its interaction with proteins. PMID- 21629647 TI - Can wild ungulate carcasses provide enough biomass to maintain avian scavenger populations? An empirical assessment using a bio-inspired computational model. AB - BACKGROUND: The reduction in the amount of food available for European avian scavengers as a consequence of restrictive public health policies is a concern for managers and conservationists. Since 2002, the application of several sanitary regulations has limited the availability of feeding resources provided by domestic carcasses, but theoretical studies assessing whether the availability of food resources provided by wild ungulates are enough to cover energetic requirements are lacking. METHODOLOGY/FINDINGS: We assessed food provided by a wild ungulate population in two areas of NE Spain inhabited by three vulture species and developed a P System computational model to assess the effects of the carrion resources provided on their population dynamics. We compared the real population trend with to a hypothetical scenario in which only food provided by wild ungulates was available. Simulation testing of the model suggests that wild ungulates constitute an important food resource in the Pyrenees and the vulture population inhabiting this area could grow if only the food provided by wild ungulates would be available. On the contrary, in the Pre-Pyrenees there is insufficient food to cover the energy requirements of avian scavenger guilds, declining sharply if biomass from domestic animals would not be available. CONCLUSIONS/SIGNIFICANCE: Our results suggest that public health legislation can modify scavenger population trends if a large number of domestic ungulate carcasses disappear from the mountains. In this case, food provided by wild ungulates could be not enough and supplementary feeding could be necessary if other alternative food resources are not available (i.e. the reintroduction of wild ungulates), preferably in European Mediterranean scenarios sharing similar and socio-economic conditions where there are low densities of wild ungulates. Managers should anticipate the conservation actions required by assessing food availability and the possible scenarios in order to make the most suitable decisions. PMID- 21629648 TI - New insight into the antifibrotic effects of praziquantel on mice in infection with Schistosoma japonicum. AB - BACKGROUND: Schistosomiasis is a parasitic disease infecting more than 200 million people in the world. Although chemotherapy targeting on killing schistosomes is one of the main strategies in the disease control, there are few effective ways of dealing with liver fibrosis caused by the parasite infection in the chronic and advanced stages of schistosomiasis. For this reason, new strategies and prospective drugs, which exert antifibrotic effects, are urgently required. METHODS AND FINDINGS: The antifibrotic effects of praziquantel were assessed in the murine models of schistosomiasis japonica. Murine fibrosis models were established by cutaneous infection with 14 +/- 2 Schistosoma japonicum cercariae. Then, the mice of both chronic (8 weeks post-infection) and advanced (15 weeks post-infection) schistosomiasis were treated by gavage of praziquantel (250 mg/kg, once daily for 3 days) to eliminate worms, and followed by praziquantel anti-fibrosis treatment (300 mg/kg, twice daily for 30 days). The fibrosis-related parameters assessed were areas of collagen deposition, content of hydroxyproline and mRNA expressions of Col1alpha1, Col3alpha1, alpha-SMA, TGF beta, MMP9, TIMP1, IL-4, IL-10, IL-13 and IFN-gamma of liver. Spleen weight index, alanine aminotransferase activity and liver portal venous pressure were also measured. The results showed that anti-fibrosis treatment improved liver fibrosis, splenomegaly, hepatic function, as well as liver portal hypertension. In order to confirm the anti-fibrotic properties of praziquantel, we established a CCL(4)-induced model and revealed that CCL(4)-induced liver fibrosis was inhibited by PZQ treatment for 30 days. Furthermore, we analyzed the effects of praziquantel on mouse primary hepatic stellate cells (HSCs). It is indicated that mRNA expressions of Col1alpha1, Col3alpha1, alpha-SMA, TGF-beta, MMP9 and TIMP1 of HSCs were all inhibited after praziquantel anti-parasite treatments. CONCLUSIONS: The significant amelioration of hepatic fibrosis by praziquantel treatment validates it as a promising drug of anti-fibrosis and offers potential of a new chemotherapy for hepatic fibrosis resulting from schistosomiasis. PMID- 21629649 TI - Climate change affects winter chill for temperate fruit and nut trees. AB - BACKGROUND: Temperate fruit and nut trees require adequate winter chill to produce economically viable yields. Global warming has the potential to reduce available winter chill and greatly impact crop yields. METHODOLOGY/PRINCIPAL FINDINGS: We estimated winter chill for two past (1975 and 2000) and 18 future scenarios (mid and end 21st century; 3 Global Climate Models [GCMs]; 3 greenhouse gas emissions [GHG] scenarios). For 4,293 weather stations around the world and GCM projections, Safe Winter Chill (SWC), the amount of winter chill that is exceeded in 90% of all years, was estimated for all scenarios using the "Dynamic Model" and interpolated globally. We found that SWC ranged between 0 and about 170 Chill Portions (CP) for all climate scenarios, but that the global distribution varied across scenarios. Warm regions are likely to experience severe reductions in available winter chill, potentially threatening production there. In contrast, SWC in most temperate growing regions is likely to remain relatively unchanged, and cold regions may even see an increase in SWC. Climate change impacts on SWC differed quantitatively among GCMs and GHG scenarios, with the highest GHG leading to losses up to 40 CP in warm regions, compared to 20 CP for the lowest GHG. CONCLUSIONS/SIGNIFICANCE: The extent of projected changes in winter chill in many major growing regions of fruits and nuts indicates that growers of these commodities will likely experience problems in the future. Mitigation of climate change through reductions in greenhouse gas emissions can help reduce the impacts, however, adaption to changes will have to occur. To better prepare for likely impacts of climate change, efforts should be undertaken to breed tree cultivars for lower chilling requirements, to develop tools to cope with insufficient winter chill, and to better understand the temperature responses of tree crops. PMID- 21629650 TI - Risk of advanced colorectal neoplasia according to age and gender. AB - BACKGROUND: Colorectal cancer (CRC) is one of the leading causes of cancer related morbidity and death. Despite the fact that the mean age at diagnosis of CRC is lower in men, screening by colonoscopy or fecal occult blood test (FOBT) is initiated at same age in both genders. The prevalence of the common CRC precursor lesion, advanced adenoma, is well documented only in the screening population. The purpose of this study was to assess the risk of advanced adenoma at ages below screening age. METHODS AND FINDINGS: We analyzed data from a census of 625,918 outpatient colonoscopies performed in adults in Bavaria between 2006 and 2008. A logistic regression model to determine gender- and age-specific risk of advanced neoplasia was developed. Advanced neoplasia was found in 16,740 women (4.6%) and 22,684 men (8.6%). Male sex was associated with an overall increased risk of advanced neoplasia (odds ratio 1.95; 95% confidence interval, CI, 1.91 to 2.00). At any age and in any indication group, more colonoscopies were needed in women than in men to detect advanced adenoma or cancer. At age 75 14.8 (95% CI, 14.4-15.2) screening, 18.2 (95% CI, 17.7-18.7) diagnostic, and 7.9 (95% CI, 7.6 8.2) colonoscopies to follow up on a positive FOBT (FOBT colonoscopies) were needed to find advanced adenoma in women. At age 50 39.0 (95% CI, 38.0-40.0) diagnostic, and 16.3 (95% CI, 15.7-16.9) FOBT colonoscopies were needed. Comparable numbers were reached 20 and 10 years earlier in men than in women, respectively. CONCLUSIONS: At any age and independent of the indication for colonoscopy, men are at higher risk of having advanced neoplasia diagnosed upon colonoscopy than women. This suggests that starting screening earlier in life in men than in women might result in a relevant increase in the detection of asymptomatic preneoplastic and neoplastic colonic lesions. PMID- 21629651 TI - The potential contribution of mass treatment to the control of Plasmodium falciparum malaria. AB - Mass treatment as a means to reducing P. falciparum malaria transmission was used during the first global malaria eradication campaign and is increasingly being considered for current control programmes. We used a previously developed mathematical transmission model to explore both the short and long-term impact of possible mass treatment strategies in different scenarios of endemic transmission. Mass treatment is predicted to provide a longer-term benefit in areas with lower malaria transmission, with reduced transmission levels for at least 2 years after mass treatment is ended in a scenario where the baseline slide-prevalence is 5%, compared to less than one year in a scenario with baseline slide-prevalence at 50%. However, repeated annual mass treatment at 80% coverage could achieve around 25% reduction in infectious bites in moderate-to high transmission settings if sustained. Using vector control could reduce transmission to levels at which mass treatment has a longer-term impact. In a limited number of settings (which have isolated transmission in small populations of 1000-10,000 with low-to-medium levels of baseline transmission) we find that five closely spaced rounds of mass treatment combined with vector control could make at least temporary elimination a feasible goal. We also estimate the effects of using gametocytocidal treatments such as primaquine and of restricting treatment to parasite-positive individuals. In conclusion, mass treatment needs to be repeated or combined with other interventions for long-term impact in many endemic settings. The benefits of mass treatment need to be carefully weighed against the risks of increasing drug selection pressure. PMID- 21629652 TI - Aging, transition, and estimating the global burden of disease. AB - BACKGROUND: The World Health Organization's Global Burden of Disease (GBD) reports are an important tool for global health policy makers, however the accuracy of estimates for countries undergoing an epidemiologic transition is unclear. We attempted to validate the life table model used to generate estimates for all-cause mortality in developing countries. METHODS AND RESULTS: Data were obtained for males and females from the Human Mortality Database for all countries with available data every ten years from 1900 to 2000. These provided inputs for the GBD life table model and served as comparison observed data. Above age sixty model estimates of survival for both sexes differed substantially from those observed. Prior to the year 1960 for males and 1930 for females, estimated survival tended to be greater than observed; following 1960 for both males and females estimated survival tended to be less than observed. Viewing observed and estimated survival separately, observed survival past sixty increased over the years considered. For males, the increase was from a mean (sd) probability of 0.22 (0.06) to 0.46 (0.1). For females, the increase was from 0.26 (0.06) to 0.65 (0.08). By contrast, estimated survival past sixty decreased over the same period. Among males, estimated survival probability declined from 0.54 (0.2) to 0.09 (0.06). Among females, the decline was from 0.36 (0.12) to 0.15 (0.08). CONCLUSIONS: These results show that the GBD mortality model did not accurately estimate survival at older ages as developed countries transitioned in the twentieth century and may be similarly flawed in developing countries now undergoing transition. Estimates of the size of older-age populations and their attributable disease burden should be reconsidered. PMID- 21629653 TI - Integrated microRNA-mRNA-analysis of human monocyte derived macrophages upon Mycobacterium avium subsp. hominissuis infection. AB - BACKGROUND: Many efforts have been made to understand basal mechanisms of mycobacterial infections. Macrophages are the first line of host immune defence to encounter and eradicate mycobacteria. Pathogenic species have evolved different mechanisms to evade host response, e.g. by influencing macrophage apoptotic pathways. However, the underlying molecular regulation is not fully understood. A new layer of eukaryotic regulation of gene expression is constituted by microRNAs. Therefore, we present a comprehensive study for identification of these key regulators and their targets in the context of host macrophage response to mycobacterial infections. METHODOLOGY/PRINCIPAL FINDINGS: We performed microRNA as well as mRNA expression analysis of human monocyte derived macrophages infected with several Mycobacterium avium hominissuis strains by means of microarrays as well as quantitative reverse transcription PCR (qRT PCR). The data revealed the ability of all strains to inhibit apoptosis by transcriptional regulation of BCL2 family members. Accordingly, at 48 h after infection macrophages infected with all M. avium strains showed significantly decreased caspase 3 and 7 activities compared to the controls. Expression of let 7e, miR-29a and miR-886-5p were increased in response to mycobacterial infection at 48 h. The integrated analysis of microRNA and mRNA expression as well as target prediction pointed out regulative networks identifying caspase 3 and 7 as potential targets of let-7e and miR-29a, respectively. Consecutive reporter assays verified the regulation of caspase 3 and 7 by these microRNAs. CONCLUSIONS/SIGNIFICANCE: We show for the first time that mycobacterial infection of human macrophages causes a specific microRNA response. We furthermore outlined a regulatory network of potential interactions between microRNAs and mRNAs. This study provides a theoretical concept for unveiling how distinct mycobacteria could manipulate host cell response. In addition, functional relevance was confirmed by uncovering the control of major caspases 3 and 7 by let-7e and miR 29a, respectively. PMID- 21629654 TI - Gradients in the number of species at reef-seagrass ecotones explained by gradients in abundance. AB - Gradients in the composition and diversity (e.g. number of species) of faunal assemblages are common at ecotones between juxtaposed habitats. Patterns in the number of species, however, can be confounded by patterns in abundance of individuals, because more species tend to be found wherever there are more individuals. We tested whether proximity to reefs influenced patterns in the composition and diversity ('species density' = number of species per area and 'species richness' = number of species per number of individuals) of prosobranch gastropods in meadows of two seagrasses with different physiognomy: Posidonia and Amphibolis. A change in the species composition was observed from reef-seagrass edges towards the interiors of Amphibolis, but not in Posidonia meadows. Similarly, the abundance of gastropods and species density was higher at edges relative to interiors of Amphibolis meadows, but not in Posidonia meadows. However, species richness was not affected by proximity to reefs in either type of seagrass meadow. The higher number of species at the reef-Amphibolis edge was therefore a consequence of higher abundance, rather than species richness per se. These results suggest that patterns in the composition and diversity of fauna with proximity to adjacent habitats, and the underlying processes that they reflect, likely depend on the physiognomy of the habitat. PMID- 21629655 TI - Genetic structure of the tiger mosquito, Aedes albopictus, in Cameroon (Central Africa). AB - BACKGROUND: Aedes albopictus (Skuse, 1884) (Diptera: Culicidae), a mosquito native to Asia, has recently invaded all five continents. In Central Africa it was first reported in the early 2000s, and has since been implicated in the emergence of arboviruses such as dengue and chikungunya in this region. Recent genetic studies of invasive species have shown that multiple introductions are a key factor for successful expansion in new areas. As a result, phenotypic characters such as vector competence and insecticide susceptibility may vary within invasive pest species, potentially affecting vector efficiency and pest management. Here we assessed the genetic variability and population genetics of Ae. albopictus isolates in Cameroon (Central Africa), thereby deducing their likely geographic origin. METHODS AND RESULTS: Mosquitoes were sampled in 2007 in 12 localities in southern Cameroon and analyzed for polymorphism at six microsatellite loci and in two mitochondrial DNA regions (ND5 and COI). All the microsatellite markers were successfully amplified and were polymorphic, showing moderate genetic structureamong geographic populations (F(ST) = 0.068, P < 0.0001). Analysis of mtDNA sequences revealed four haplotypes each for the COI and ND5 genes, with a dominant haplotype shared by all Cameroonian samples. The weak genetic variation estimated from the mtDNA genes is consistent with the recent arrival of Ae. albopictus in Cameroon. Phylogeographic analysis based on COI polymorphism indicated that Ae. albopictus populations from Cameroon are related to tropical rather than temperate or subtropical outgroups. CONCLUSION: The moderate genetic diversity observed among Cameroonian Ae. albopictus isolates is in keeping with recent introduction and spread in this country. The genetic structure of natural populations points to multiple introductions from tropical regions. PMID- 21629656 TI - Enhanced neointima formation following arterial injury in immune deficient Rag-1 /- mice is attenuated by adoptive transfer of CD8 T cells. AB - T cells modulate neointima formation after arterial injury but the specific T cell population that is activated in response to arterial injury remains unknown. The objective of the study was to identify the T cell populations that are activated and modulate neointimal thickening after arterial injury in mice. Arterial injury in wild type C57Bl6 mice resulted in T cell activation characterized by increased CD4(+)CD44(hi) and CD8(+)CD44(hi) T cells in the lymph nodes and spleens. Splenic CD8(+)CD25(+) T cells and CD8(+)CD28(+) T cells, but not CD4(+)CD25(+) and CD4(+)CD28(+) T cells, were also significantly increased. Adoptive cell transfer of CD4(+) or CD8(+) T cells from donor CD8-/- or CD4-/- mice, respectively, to immune-deficient Rag-1-/- mice was performed to determine the T cell subtype that inhibits neointima formation after arterial injury. Rag-1 /- mice that received CD8(+) T cells had significantly reduced neointima formation compared with Rag-1-/- mice without cell transfer. CD4(+) T cell transfer did not reduce neointima formation. CD8(+) T cells from CD4-/- mice had cytotoxic activity against syngeneic smooth muscle cells in vitro. The study shows that although both CD8(+) T cells and CD4(+) T cells are activated in response to arterial injury, adoptive cell transfer identifies CD8(+) T cells as the specific and selective cell type involved in inhibiting neointima formation. PMID- 21629657 TI - Omeprazole inhibits proliferation and modulates autophagy in pancreatic cancer cells. AB - BACKGROUND: Omeprazole has recently been described as a modulator of tumour chemoresistance, although its underlying molecular mechanisms remain controversial. Since pancreatic tumours are highly chemoresistant, a logical step would be to investigate the pharmacodynamic, morphological and biochemical effects of omeprazole on pancreatic cancer cell lines. METHODOLOGY/PRINCIPAL FINDINGS: Dose-effect curves of omeprazole, pantoprazole, gemcitabine, 5 fluorouracil and the combinations of omeprazole and 5-fluorouracil or gemcitabine were generated for the pancreatic cancer cell lines MiaPaCa-2, ASPC-1, Colo357, PancTu-1, Panc1 and Panc89. They revealed that omeprazole inhibited proliferation at probably non-toxic concentrations and reversed the hormesis phenomena of 5 fluorouracil. Electron microscopy showed that omeprazole led to accumulation of phagophores and early autophagosomes in ASPC-1 and MiaPaCa-2 cells. Signal changes indicating inhibited proliferation and programmed cell death were found by proton NMR spectroscopy of both cell lines when treated with omeprazole which was identified intracellularly. Omeprazole modulates the lysosomal transport pathway as shown by Western blot analysis of the expression of LAMP-1, Cathepsin D and beta-COP in lysosome- and Golgi complex containing cell fractions. Acridine orange staining revealed that the pump function of the vATPase was not specifically inhibited by omeprazole. Gene expression of the autophagy-related LC3 gene as well as of Bad, Mdr-1, Atg12 and the vATPase was analysed after treatment of cells with 5-fluorouracil and omeprazole and confirmed the above mentioned results. CONCLUSIONS: We hypothesise that omeprazole interacts with the regulatory functions of the vATPase without inhibiting its pump function. A modulation of the lysosomal transport pathway and autophagy is caused in pancreatic cancer cells leading to programmed cell death. This may circumvent common resistance mechanisms of pancreatic cancer. Since omeprazole use has already been established in clinical practice these results could lead to new clinical applications. PMID- 21629658 TI - Calbindin 2 (CALB2) regulates 5-fluorouracil sensitivity in colorectal cancer by modulating the intrinsic apoptotic pathway. AB - The role of the calcium binding protein, Calbindin 2 (CALB2), in regulating the response of colorectal cancer (CRC) cells to 5-Fluorouracil (5-FU) was investigated. Real-time RT-PCR and Western blot analysis revealed that CALB2 mRNA and protein expression were down-regulated in p53 wild-type and p53 null isogenic HCT116 CRC cell lines following 48 h and 72 h 5-FU treatment. Moreover, 5-FU induced apoptosis was significantly reduced in HCT116 and LS174T CRC cell lines in which CALB2 expression had been silenced. Further investigation revealed that CALB2 translocated to the mitochondria following 5-FU treatment and that 5-FU induced loss of mitochondrial membrane potential (Deltapsi(m)) was abrogated in CALB2-silenced cells. Furthermore, CALB2 silencing decreased 5-FU-induced cytochrome c and smac release from the mitochondria and also decreased 5-FU induced activation of caspases 9 and 3/7. Of note, co-silencing of XIAP overcame 5-FU resistance in CALB2-silenced cells. Collectively, these results suggest that following 5-FU treatment in CRC cell lines, CALB2 is involved in apoptosis induction through the intrinsic mitochondrial pathway. This indicates that CALB2 may be an important mediator of 5-FU-induced cell death. Moreover, down regulation of CALB2 in response to 5-FU may represent an intrinsic mechanism of resistance to this anti-cancer drug. PMID- 21629659 TI - Measuring generalization of visuomotor perturbations in wrist movements using mobile phones. AB - Recent studies in motor control have shown that visuomotor rotations for reaching have narrow generalization functions: what we learn during movements in one direction only affects subsequent movements into close directions. Here we wanted to measure the generalization functions for wrist movement. To do so we had 7 subjects performing an experiment holding a mobile phone in their dominant hand. The mobile phone's built in acceleration sensor provided a convenient way to measure wrist movements and to run the behavioral protocol. Subjects moved a cursor on the screen by tilting the phone. Movements on the screen toward the training target were rotated and we then measured how learning of the rotation in the training direction affected subsequent movements in other directions. We find that generalization is local and similar to generalization patterns of visuomotor rotation for reaching. PMID- 21629660 TI - Diarrhea, pneumonia, and infectious disease mortality in children aged 5 to 14 years in India. AB - BACKGROUND: Little is known about the causes of death in children in India after age five years. The objective of this study is to provide the first ever direct national and sub-national estimates of infectious disease mortality in Indian children aged 5 to 14 years. METHODS: A verbal autopsy based assessment of 3 855 deaths is children aged 5 to 14 years from a nationally representative survey of deaths occurring in 2001-03 in 1.1 million homes in India. RESULTS: Infectious diseases accounted for 58% of all deaths among children aged 5 to 14 years. About 18% of deaths were due to diarrheal diseases, 10% due to pneumonia, 8% due to central nervous system infections, 4% due to measles, and 12% due to other infectious diseases. Nationally, in 2005 about 59 000 and 34 000 children aged 5 to 14 years died from diarrheal diseases and pneumonia, corresponding to mortality of 24.1 and 13.9 per 100 000 respectively. Mortality was nearly 50% higher in girls than in boys for both diarrheal diseases and pneumonia. CONCLUSIONS: Approximately 60% of all deaths in this age group are due to infectious diseases and nearly half of these deaths are due to diarrheal diseases and pneumonia. Mortality in this age group from infectious diseases, and diarrhea in particular, is much higher than previously estimated. PMID- 21629661 TI - Probenecid inhibits the human bitter taste receptor TAS2R16 and suppresses bitter perception of salicin. AB - Bitter taste stimuli are detected by a diverse family of G protein-coupled receptors (GPCRs) expressed in gustatory cells. Each bitter taste receptor (TAS2R) responds to an array of compounds, many of which are toxic and can be found in nature. For example, human TAS2R16 (hTAS2R16) responds to beta glucosides such as salicin, and hTAS2R38 responds to thiourea-containing molecules such as glucosinolates and phenylthiocarbamide (PTC). While many substances are known to activate TAS2Rs, only one inhibitor that specifically blocks bitter receptor activation has been described. Here, we describe a new inhibitor of bitter taste receptors, p-(dipropylsulfamoyl)benzoic acid (probenecid), that acts on a subset of TAS2Rs and inhibits through a novel, allosteric mechanism of action. Probenecid is an FDA-approved inhibitor of the Multidrug Resistance Protein 1 (MRP1) transporter and is clinically used to treat gout in humans. Probenecid is also commonly used to enhance cellular signals in GPCR calcium mobilization assays. We show that probenecid specifically inhibits the cellular response mediated by the bitter taste receptor hTAS2R16 and provide molecular and pharmacological evidence for direct interaction with this GPCR using a non-competitive (allosteric) mechanism. Through a comprehensive analysis of hTAS2R16 point mutants, we define amino acid residues involved in the probenecid interaction that result in decreased sensitivity to probenecid while maintaining normal responses to salicin. Probenecid inhibits hTAS2R16, hTAS2R38, and hTAS2R43, but does not inhibit the bitter receptor hTAS2R31 or non-TAS2R GPCRs. Additionally, structurally unrelated MRP1 inhibitors, such as indomethacin, fail to inhibit hTAS2R16 function. Finally, we demonstrate that the inhibitory activity of probenecid in cellular experiments translates to inhibition of bitter taste perception of salicin in humans. This work identifies probenecid as a pharmacological tool for understanding the cell biology of bitter taste and as a lead for the development of broad specificity bitter blockers to improve nutrition and medical compliance. PMID- 21629662 TI - Determination of the molecular basis for a limited dimorphism, N417K, in the Plasmodium vivax Duffy-binding protein. AB - Invasion of human red blood cells by Plasmodium merozoites is vital for replication and survival of the parasite and, as such, is an attractive target for therapeutic intervention. Merozoite invasion is mediated by specific interactions between parasite ligands and host erythrocyte receptors. The P. vivax Duffy-binding protein (PvDBP) is heavily dependent on the interaction with the human Duffy blood group antigen/receptor for chemokines (DARC) for invasion. Region II of PvDBP contains many allelic polymorphisms likely to have arisen by host immune selection. Successful vaccine development necessitates a deeper understanding of the role of these polymorphisms in both parasite function and evasion of host immunity. A 3D structure of the homologous P. knowlesi DBP predicts that most variant residues are surface-exposed, including N417K, which is a dimorphic residue change that has previously been shown to be part of a linked haplotype that alters DBP sensitivity to inhibitory antibody. In natural isolates only two residues are found at this site, asparagine (N) and lysine (K). Site-directed mutagenesis of residue 417 was used to create a panel of 20 amino acid variants that were then examined for their binding phenotype and response to immune sera. Our results suggest that the observed dimorphism likely arose due to both structural requirements and immune selection pressure. To our knowledge, this is the first exhaustive examination of this kind of the role of a single amino acid residue in antigenic character and binding ability. Our results demonstrate that a single amino acid substitution can dramatically alter both the ability of the PvDBP to bind to human erythrocytes and its antigenic character. PMID- 21629663 TI - Accelerated neuronal cell recovery from Botulinum neurotoxin intoxication by targeted ubiquitination. AB - Botulinum neurotoxin (BoNT), a Category A biodefense agent, delivers a protease to motor neuron cytosol that cleaves one or more soluble NSF attachment protein receptors (SNARE) proteins involved in neurotransmission to cause a flaccid paralysis. No antidotes exist to reverse symptoms of BoNT intoxication so severely affected patients require artificial respiration with prolonged intensive care. Time to recovery depends on toxin serotype because the intraneuronal persistence of the seven known BoNT serotypes varies widely from days to many months. Our therapeutic antidote strategy is to develop 'targeted F box' (TFB) agents that target the different intraneuronal BoNT proteases for accelerated degradation by the ubiquitin proteasome system (UPS), thus promoting rapid recovery from all serotypes. These agents consist of a camelid heavy chain only V(H) (VHH) domain specific for a BoNT protease fused to an F-box domain recognized by an intraneuronal E3-ligase. A fusion protein containing the 14 kDa anti-BoNT/A protease VHH, ALcB8, joined to a 15 kDa F-box domain region of TrCP (D5) was sufficient to cause increased ubiquitination and accelerate turnover of the targeted BoNT/A protease within neurons. Neuronal cells expressing this TFB, called D5-B8, were also substantially resistant to BoNT/A intoxication and recovered from intoxication at least 2.5 fold quicker than control neurons. Fusion of D5 to a VHH specific for BoNT/B protease (BLcB10) led to accelerated turnover of the targeted protease within neurons, thus demonstrating the modular nature of these therapeutic agents and suggesting that development of similar therapeutic agents specific to all botulinum serotypes should be readily achievable. PMID- 21629664 TI - Going deeper: metagenome of a hadopelagic microbial community. AB - The paucity of sequence data from pelagic deep-ocean microbial assemblages has severely restricted molecular exploration of the largest biome on Earth. In this study, an analysis is presented of a large-scale 454-pyrosequencing metagenomic dataset from a hadopelagic environment from 6,000 m depth within the Puerto Rico Trench (PRT). A total of 145 Mbp of assembled sequence data was generated and compared to two pelagic deep ocean metagenomes and two representative surface seawater datasets from the Sargasso Sea. In a number of instances, all three deep metagenomes displayed similar trends, but were most magnified in the PRT, including enrichment in functions for two-component signal transduction mechanisms and transcriptional regulation. Overrepresented transporters in the PRT metagenome included outer membrane porins, diverse cation transporters, and di- and tri-carboxylate transporters that matched well with the prevailing catabolic processes such as butanoate, glyoxylate and dicarboxylate metabolism. A surprisingly high abundance of sulfatases for the degradation of sulfated polysaccharides were also present in the PRT. The most dramatic adaptational feature of the PRT microbes appears to be heavy metal resistance, as reflected in the large numbers of transporters present for their removal. As a complement to the metagenome approach, single-cell genomic techniques were utilized to generate partial whole-genome sequence data from four uncultivated cells from members of the dominant phyla within the PRT, Alphaproteobacteria, Gammaproteobacteria, Bacteroidetes and Planctomycetes. The single-cell sequence data provided genomic context for many of the highly abundant functional attributes identified from the PRT metagenome, as well as recruiting heavily the PRT metagenomic sequence data compared to 172 available reference marine genomes. Through these multifaceted sequence approaches, new insights have been provided into the unique functional attributes present in microbes residing in a deeper layer of the ocean far removed from the more productive sun-drenched zones above. PMID- 21629665 TI - Sphingosine 1-phosphate induces differentiation of mesoangioblasts towards smooth muscle. A role for GATA6. AB - Different cells can contribute to repair following vascular injury by differentiating into smooth muscle (SM) cells; however the extracellular signals involved are presently poorly characterized. Mesoangioblasts are progenitor cells capable of differentiating into various mesoderm cell types including SM cells. In this study the biological action exerted by the pleiotropic sphingolipid sphingosine 1-phosphate (S1P) in human mesoangioblasts has been initially investigated by cDNA microarray analysis. Obtained data confirmed the anti apoptotic action of this sphingolipid and identified for the first time a strong differentiating action toward SM cells. Quantitative mRNA and protein analysis corroborated the microarray results demonstrating enhanced expression of myogenic marker proteins and regulation of the expression of transcription factor GATA6 and its co-regulator, LMCD1. Importantly, GATA6 up-regulation induced by S1P was responsible for the enhanced expression of SM-specific contractile proteins. Moreover, by specific gene silencing experiments GATA6 was critical in the pro differentiating activity of the cytokine TGFbeta. Finally, the pharmacological inhibition of endogenous S1P formation in response to TGFbeta abrogated GATA6 up regulation, supporting the view that the S1P pathway plays a physiological role in mediating the pro-myogenic effect of TGFbeta. This study individuates GATA6 as novel player in the complex transcriptional regulation of mesoangioblast differentiation into SM cells and highlights a role for S1P to favour vascular regeneration. PMID- 21629666 TI - VPS29 is not an active metallo-phosphatase but is a rigid scaffold required for retromer interaction with accessory proteins. AB - VPS29 is a key component of the cargo-binding core complex of retromer, a protein assembly with diverse roles in transport of receptors within the endosomal system. VPS29 has a fold related to metal-binding phosphatases and mediates interactions between retromer and other regulatory proteins. In this study we examine the functional interactions of mammalian VPS29, using X-ray crystallography and NMR spectroscopy. We find that although VPS29 can coordinate metal ions Mn(2+) and Zn(2+) in both the putative active site and at other locations, the affinity for metals is low, and lack of activity in phosphatase assays using a putative peptide substrate support the conclusion that VPS29 is not a functional metalloenzyme. There is evidence that structural elements of VPS29 critical for binding the retromer subunit VPS35 may undergo both metal dependent and independent conformational changes regulating complex formation, however studies using ITC and NMR residual dipolar coupling (RDC) measurements show that this is not the case. Finally, NMR chemical shift mapping indicates that VPS29 is able to associate with SNX1 via a conserved hydrophobic surface, but with a low affinity that suggests additional interactions will be required to stabilise the complex in vivo. Our conclusion is that VPS29 is a metal ion independent, rigid scaffolding domain, which is essential but not sufficient for incorporation of retromer into functional endosomal transport assemblies. PMID- 21629667 TI - In vitro and in vivo germ line potential of stem cells derived from newborn mouse skin. AB - We previously reported that fetal porcine skin-derived stem cells were capable of differentiation into oocyte-like cells (OLCs). Here we report that newborn mice skin-derived stem cells are also capable of differentiating into early OLCs. Using stem cells from mice that are transgenic for Oct4 germline distal enhancer GFP, germ cells resulting from their differentiation are expected to be GFP(+). After differentiation, some GFP(+) OLCs reached 40-45 uM and expressed oocyte markers. Flow cytometric analysis revealed that ~ 0.3% of the freshly isolated skin cells were GFP(+). The GFP-positive cells increased to ~ 7% after differentiation, suggesting that the GFP(+) cells could be of in vivo origin, but are more likely induced upon being cultured in vitro. To study the in vivo germ cell potential of skin-derived cells, they were aggregated with newborn ovarian cells, and transplanted under the kidney capsule of ovariectomized mice. GFP(+) oocytes were identified within a subpopulation of follicles in the resulting growth. Our finding that early oocytes can be differentiated from mice skin derived cells in defined medium may offer a new in vitro model to study germ cell formation and oogenesis. PMID- 21629668 TI - Adenosine A(2A) Receptor and IL-10 in Peripheral Blood Mononuclear Cells of Patients with Mild Cognitive Impairment. AB - Adenosine suppresses immune responses through the A(2A) receptor (A(2A)R). This study investigated the interleukin 10 (IL-10) genetic profile and the expression of A(2A)R in peripheral blood mononuclear cells (PBMCs) of patients with mild cognitive impairment (MCI), Alzheimer disease (AD), and age-matched controls to verify, if they may help distinguish different forms of cognitive decline. We analyzed the IL-10 genotype and the expression of A(2A)R in 41 subjects with AD, 10 with amnestic MCI (a-MCI), 49 with multiple cognitive domain MCI (mcd-MCI), and 46 controls. There was a significant linear increase in A(2A)R mRNA levels and A(2A)R density from mcd-MCI to a-MCI, with intermediate levels being found in AD. The IL-10 AA genotype frequency was 67% in a-MCI, 46% in AD, 35% in mcd-MCI, and 20% in controls. These data suggest that the assessment of the IL-10 genotype and the expression of A(2A)R in PBMCs may be a valuable means of differentiating between a-MCI and mcd-MCI. PMID- 21629669 TI - Faculty verbal evaluations reveal strategies used to promote medical student performance. AB - BACKGROUND: Preceptors rarely follow medical students' developing clinical performance over time and across disciplines. This study analyzes preceptors' descriptions of longitudinal integrated clerkship (LIC) students' clinical development and their identification of strategies to guide students' progress. METHODS: We used a common evaluation framework, reporter-interpreter-manager educator, to guide multidisciplinary LIC preceptors' discussions of students' progress. We conducted thematic analysis of transcripts from preceptors' (seven longitudinal ambulatory preceptors per student) quarterly group discussions of 15 students' performance over one year. RESULTS: All students' clinical development progressed, although most experienced obstacles. Lack of structure in the history and physical exam commonly obstructed progression. Preceptors used templates for data gathering, and modeling or experiences in the inpatient setting to provide time and solidify structure. To advance students' knowledge acquisition, many preceptors identified focused learning topics with their students; to promote application of knowledge, preceptors used reasoning strategies to teach the steps involved in synthesizing clinical data. Preceptors shared accountability for helping students advance as the LIC allowed them to follow students' response to teaching strategies. DISCUSSION: These results depict preceptors' perceptions of LIC students' developmental continuum and illustrate how multidisciplinary preceptors can use a common evaluation framework to identify strategies to improve performance and follow students' performance longitudinally. PMID- 21629670 TI - Novel mutation in the AVPR2 gene in a Danish male with nephrogenic diabetes insipidus caused by ER retention and subsequent lysosomal degradation of the mutant receptor. AB - Mutations in the arginine vasopressin receptor 2 (AVPR2) gene can cause X-linked nephrogenic diabetes insipidus (NDI) characterized by the production of large amounts of urine and an inability to concentrate urine in response to the antidiuretic hormone vasopressin. We have identified a novel mutation in the AVPR2 gene (L170P) located in the fourth transmembrane domain in a Danish NDI male. Analysis of the mutant receptor in Madin-Darby Canine Kidney cell culture revealed that AVPR2-L170P was retained in the endoplasmic reticulum, and the expression was dramatically downregulated compared to wild-type AVPR2. Inhibition of the lysosome resulted in increased intracellular accumulation of AVPR2-L170P, indicating that AVPR2-L170P is downregulated via the lysosome. Inhibition of the proteasome resulted in plasma membrane localization of AVPR2-L170P, although the overall levels of AVPR2-L170P were unchanged. PMID- 21629671 TI - Potential role of cathepsin K in the pathophysiology of mucopolysaccharidoses. AB - Cathepsin K, a papain-like cysteine protease, is highly expressed in osteoclasts and plays a critical role in bone resorption. Dysfunction of the enzyme leads to various skeletal abnormalities. The recent knowledge that the collagenolytic activity of cathepsin K depends on interactions with bone and cartilage-resident glycosaminoglycans (GAGs) may shed some light on diseases such as mucopolysaccharidoses (MPSs). MPSs are a group of lysosomal storage diseases characterized by the accumulation of GAGs in tissues including bone. Typical pathological features of these diseases include skeletal abnormalities such as dysostosis multiplex, short stature, and multiple irregularities in bone development. We describe how further investigation of the cathepsin K/GAG complexes could provide valuable insights into the bone pathology associated with MPS diseases. In this review, we discuss the inhibition of osteoclast function through altered activity of cathepsin K by GAGs and offer insight into a mechanism for the bone pathology seen in MPS patients. PMID- 21629672 TI - New trends in biomedical signal processing: a crossroads between smart sensors in e-health and virtual physiological human initiatives. PMID- 21629673 TI - Development of an optimised application protocol for sonophoretic transdermal delivery of a model hydrophilic drug. AB - It has now been known for over a decade that low frequency ultrasound can be used to effectively enhance transdermal drug penetration - an approach termed sonophoresis. Mechanistically, acoustic cavitation results in the creation of defects in the stratum corneum that allow accelerated absorption of topically applied molecules. The aim of this study was to develop an optimised sonophoresis protocol for studying transdermal drug delivery in vitro. To this end, caffeine was selected as a model hydrophilic drug while porcine skin was used as a model barrier. Following acoustic validation, 20kHz ultrasound was applied for different durations (range: 5 s to 10 min) using three different modes (10%, 33% or 100% duty cycles) and two distinct sonication procedures (either before or concurrent with drug deposition). Each ultrasonic protocol was assessed in terms of its heating and caffeine flux-enhancing effects. It was found that the best regimen was a concurrent 5 min, pulsed (10% duty cycle) beam of SATA intensity 0.37 W/cm(2). A key insight was that in the case of pulsed beams of 10% duty cycle, sonication concurrent with drug deposition was superior to sonication prior to drug deposition and potential mechanisms for this are discussed. PMID- 21629674 TI - Hydrogen peroxide promotes injury-induced peripheral sensory axon regeneration in the zebrafish skin. AB - Functional recovery from cutaneous injury requires not only the healing and regeneration of skin cells but also reinnervation of the skin by somatosensory peripheral axon endings. To investigate how sensory axon regeneration and wound healing are coordinated, we amputated the caudal fins of zebrafish larvae and imaged somatosensory axon behavior. Fin amputation strongly promoted the regeneration of nearby sensory axons, an effect that could be mimicked by ablating a few keratinocytes anywhere in the body. Since injury produces the reactive oxygen species hydrogen peroxide (H(2)O(2)) near wounds, we tested whether H(2)O(2) influences cutaneous axon regeneration. Exposure of zebrafish larvae to sublethal levels of exogenous H(2)O(2) promoted growth of severed axons in the absence of keratinocyte injury, and inhibiting H(2)O(2) production blocked the axon growth-promoting effects of fin amputation and keratinocyte ablation. Thus, H(2)O(2) signaling helps coordinate wound healing with peripheral sensory axon reinnervation of the skin. PMID- 21629675 TI - Rarity, species richness, and the threat of extinction--are plants the same as animals? AB - Assessment of conservation status is done both for areas or habitats and for species (or taxa). IUCN Red List categories have been the principal method of categorising species in terms of extinction risk, and have been shown to be robust and helpful in the groups for which they have been developed. A recent study highlights properties associated with extinction risk in flowering plants, focusing on the species-rich hot spot of the Cape region of South Africa, and concludes that merely following methods derived from studies of vertebrates may not provide the best estimates of extinction risk for plants. Biology, geography, and history all are important factors in risk, and the study poses many questions about how we categorise and assess species for conservation priorities. PMID- 21629676 TI - Trapping conformational states along ligand-binding dynamics of peptide deformylase: the impact of induced fit on enzyme catalysis. AB - For several decades, molecular recognition has been considered one of the most fundamental processes in biochemistry. For enzymes, substrate binding is often coupled to conformational changes that alter the local environment of the active site to align the reactive groups for efficient catalysis and to reach the transition state. Adaptive substrate recognition is a well-known concept; however, it has been poorly characterized at a structural level because of its dynamic nature. Here, we provide a detailed mechanism for an induced-fit process at atomic resolution. We take advantage of a slow, tight binding inhibitor-enzyme system, actinonin-peptide deformylase. Crystal structures of the initial open state and final closed state were solved, as well as those of several intermediate mimics captured during the process. Ligand-induced reshaping of a hydrophobic pocket drives closure of the active site, which is finally "zipped up" by additional binding interactions. Together with biochemical analyses, these data allow a coherent reconstruction of the sequence of events leading from the encounter complex to the key-lock binding state of the enzyme. A "movie" that reconstructs this entire process can be further extrapolated to catalysis. PMID- 21629677 TI - A synthetic HIV-1 subtype C backbone generates comparable PR and RT resistance profiles to a subtype B backbone in a recombinant virus assay. AB - In order to determine phenotypic protease and reverse transcriptase inhibitor associated resistance in HIV subtype C virus, we have synthetically constructed an HIV-1 subtype C (HIV-1-C) viral backbone for use in a recombinant virus assay. The in silico designed viral genome was divided into 4 fragments, which were chemically synthesized and joined together by conventional subcloning. Subsequently, gag-protease-reverse-transcriptase (GPRT) fragments from 8 HIV-1 subtype C-infected patient samples were RT-PCR-amplified and cloned into the HIV 1-C backbone (deleted for GPRT) using In-Fusion reagents. Recombinant viruses (1 to 5 per patient sample) were produced in MT4-eGFP cells where cyto-pathogenic effect (CPE), p24 and Viral Load (VL) were monitored. The resulting HIV-1-C recombinant virus stocks (RVS) were added to MT4-eGFP cells in the presence of serial dilutions of antiretroviral drugs (PI, NNRTI, NRTI) to determine the fold change in IC50 compared to the IC50 of wild-type HIV-1 virus. Additionally, viral RNA was extracted from the HIV-1-C RVS and the amplified GPRT products were used to generate recombinant virus in a subtype B backbone. Phenotypic resistance profiles in a subtype B and subtype C backbone were compared. The following observations were made: i) functional, infectious HIV-1 subtype C viruses were generated, confirmed by VL and p24 measurements; ii) their rate of infection was slower than viruses generated in the subtype B backbone; iii) they did not produce clear CPE in MT4 cells; and iv) drug resistance profiles generated in both backbones were very similar, including re-sensitizing effects like M184V on AZT. PMID- 21629678 TI - Extinction risk and diversification are linked in a plant biodiversity hotspot. AB - It is widely recognized that we are entering an extinction event on a scale approaching the mass extinctions seen in the fossil record. Present-day rates of extinction are estimated to be several orders of magnitude greater than background rates and are projected to increase further if current trends continue. In vertebrates, species traits, such as body size, fecundity, and geographic range, are important predictors of vulnerability. Although plants are the basis for life on Earth, our knowledge of plant extinctions and vulnerabilities is lagging. Here, we disentangle the underlying drivers of extinction risk in plants, focusing on the Cape of South Africa, a global biodiversity hotspot. By comparing Red List data for the British and South African floras, we demonstrate that the taxonomic distribution of extinction risk differs significantly between regions, inconsistent with a simple, trait-based model of extinction. Using a comprehensive phylogenetic tree for the Cape, we reveal a phylogenetic signal in the distribution of plant extinction risks but show that the most threatened species cluster within short branches at the tips of the phylogeny--opposite to trends in mammals. From analyzing the distribution of threatened species across 11 exemplar clades, we suggest that mode of speciation best explains the unusual phylogenetic structure of extinction risks in plants of the Cape. Our results demonstrate that explanations for elevated extinction risk in plants of the Cape flora differ dramatically from those recognized for vertebrates. In the Cape, extinction risk is higher for young and fast-evolving plant lineages and cannot be explained by correlations with simple biological traits. Critically, we find that the most vulnerable plant species are nonetheless marching towards extinction at a more rapid pace but, surprisingly, independently from anthropogenic effects. Our results have important implications for conservation priorities and cast doubts on the utility of current Red List criteria for plants in regions such as the Cape, where speciation has been rapid, if our aim is to maximize the preservation of the tree-of-life. PMID- 21629679 TI - Health aspects of the pre-departure phase of migration. PMID- 21629680 TI - Impact of visual repetition rate on intrinsic properties of low frequency fluctuations in the visual network. AB - BACKGROUND: Visual processing network is one of the functional networks which have been reliably identified to consistently exist in human resting brains. In our work, we focused on this network and investigated the intrinsic properties of low frequency (0.01-0.08 Hz) fluctuations (LFFs) during changes of visual stimuli. There were two main questions to be discussed in this study: intrinsic properties of LFFs regarding (1) interactions between visual stimuli and resting state; (2) impact of repetition rate of visual stimuli. METHODOLOGY/PRINCIPAL FINDINGS: We analyzed scanning sessions that contained rest and visual stimuli in various repetition rates with a novel method. The method included three numerical approaches involving ICA (Independent Component Analyses), fALFF (fractional Amplitude of Low Frequency Fluctuation), and Coherence, to respectively investigate the modulations of visual network pattern, low frequency fluctuation power, and interregional functional connectivity during changes of visual stimuli. We discovered when resting-state was replaced by visual stimuli, more areas were involved in visual processing, and both stronger low frequency fluctuations and higher interregional functional connectivity occurred in visual network. With changes of visual repetition rate, the number of areas which were involved in visual processing, low frequency fluctuation power, and interregional functional connectivity in this network were also modulated. CONCLUSIONS/SIGNIFICANCE: To combine the results of prior literatures and our discoveries, intrinsic properties of LFFs in visual network are altered not only by modulations of endogenous factors (eye-open or eye-closed condition; alcohol administration) and disordered behaviors (early blind), but also exogenous sensory stimuli (visual stimuli with various repetition rates). It demonstrates that the intrinsic properties of LFFs are valuable to represent physiological states of human brains. PMID- 21629681 TI - Migration and health: a framework for 21st century policy-making. PMID- 21629682 TI - Paraoxonase 1 (PON1) polymorphisms, haplotypes and activity in predicting cad risk in North-West Indian Punjabis. AB - BACKGROUND: Human serum paraoxonase-1 (PON1) prevents oxidation of low density lipoprotein cholesterol (LDL-C) and hydrolyzes the oxidized form, therefore preventing the development of atherosclerosis. The polymorphisms of PON1 gene are known to affect the PON1 activity and thereby coronary artery disease (CAD) risk. As studies are lacking in North-West Indian Punjabi's, a distinct ethnic group with high incidence of CAD, we determined PON1 activity, genotypes and haplotypes in this population and correlated them with the risk of CAD. METHODOLOGY/PRINCIPAL FINDINGS: 350 angiographically proven (>= 70% stenosis) CAD patients and 300 healthy controls were investigated. PON1 activity was determined towards paraoxon (Paraoxonase; PONase) and phenylacetate (Arylesterase; AREase) substrates. In addition, genotyping was carried out by using multiplex PCR, allele specific oligonucleotide -PCR and PCR-RFLP methods and haplotyping was determined by PHASE software. The serum PONase and AREase activities were significantly lower in CAD patients as compared to the controls. All studied polymorphisms except L55M had significant effect on PONase activity. However AREase activity was not affected by them. In a logistic regression model, after adjustment for the conventional risk factors for CAD, QR (OR: 2.73 (1.57-4.72)) and RR (OR, 16.24 (6.41-41.14)) genotypes of Q192R polymorphism and GG (OR: 2.07 (1.02-4.21)) genotype of -162A/G polymorphism had significantly higher CAD risk. Haplotypes L-T-G-Q-C (OR: 3.25 (1.72-6.16)) and L-T-G-R-G (OR: 2.82 (1.01-7.80)) were also significantly associated with CAD. CONCLUSIONS: In conclusion this study shows that CAD patients had lower PONase and AREase activities as compared to the controls. The coding Q192R polymorphism, promoter -162A/G polymorphism and L-T-G-Q-C and L-T-G-R-G haplotypes are all independently associated with CAD. PMID- 21629684 TI - A Bayesian framework for parameter estimation in dynamical models. AB - Mathematical models in biology are powerful tools for the study and exploration of complex dynamics. Nevertheless, bringing theoretical results to an agreement with experimental observations involves acknowledging a great deal of uncertainty intrinsic to our theoretical representation of a real system. Proper handling of such uncertainties is key to the successful usage of models to predict experimental or field observations. This problem has been addressed over the years by many tools for model calibration and parameter estimation. In this article we present a general framework for uncertainty analysis and parameter estimation that is designed to handle uncertainties associated with the modeling of dynamic biological systems while remaining agnostic as to the type of model used. We apply the framework to fit an SIR-like influenza transmission model to 7 years of incidence data in three European countries: Belgium, the Netherlands and Portugal. PMID- 21629683 TI - Characterizing the epidemiology of the 2009 influenza A/H1N1 pandemic in Mexico. AB - BACKGROUND: Mexico's local and national authorities initiated an intense public health response during the early stages of the 2009 A/H1N1 pandemic. In this study we analyzed the epidemiological patterns of the pandemic during April December 2009 in Mexico and evaluated the impact of nonmedical interventions, school cycles, and demographic factors on influenza transmission. METHODS AND FINDINGS: We used influenza surveillance data compiled by the Mexican Institute for Social Security, representing 40% of the population, to study patterns in influenza-like illness (ILIs) hospitalizations, deaths, and case-fatality rate by pandemic wave and geographical region. We also estimated the reproduction number (R) on the basis of the growth rate of daily cases, and used a transmission model to evaluate the effectiveness of mitigation strategies initiated during the spring pandemic wave. A total of 117,626 ILI cases were identified during April December 2009, of which 30.6% were tested for influenza, and 23.3% were positive for the influenza A/H1N1 pandemic virus. A three-wave pandemic profile was identified, with an initial wave in April-May (Mexico City area), a second wave in June-July (southeastern states), and a geographically widespread third wave in August-December. The median age of laboratory confirmed ILI cases was ~ 18 years overall and increased to ~ 31 years during autumn (p<0.0001). The case-fatality ratio among ILI cases was 1.2% overall, and highest (5.5%) among people over 60 years. The regional R estimates were 1.8-2.1, 1.6-1.9, and 1.2-1.3 for the spring, summer, and fall waves, respectively. We estimate that the 18-day period of mandatory school closures and other social distancing measures implemented in the greater Mexico City area was associated with a 29%-37% reduction in influenza transmission in spring 2009. In addition, an increase in R was observed in late May and early June in the southeast states, after mandatory school suspension resumed and before summer vacation started. State-specific fall pandemic waves began 2-5 weeks after school reopened for the fall term, coinciding with an age shift in influenza cases. CONCLUSIONS: We documented three spatially heterogeneous waves of the 2009 A/H1N1 pandemic virus in Mexico, which were characterized by a relatively young age distribution of cases. Our study highlights the importance of school cycles on the transmission dynamics of this pandemic influenza strain and suggests that school closure and other mitigation measures could be useful to mitigate future influenza pandemics. Please see later in the article for the Editors' Summary. PMID- 21629685 TI - Medical students' exposure to and attitudes about the pharmaceutical industry: a systematic review. AB - BACKGROUND: The relationship between health professionals and the pharmaceutical industry has become a source of controversy. Physicians' attitudes towards the industry can form early in their careers, but little is known about this key stage of development. METHODS AND FINDINGS: We performed a systematic review reported according to PRISMA guidelines to determine the frequency and nature of medical students' exposure to the drug industry, as well as students' attitudes concerning pharmaceutical policy issues. We searched MEDLINE, EMBASE, Web of Science, and ERIC from the earliest available dates through May 2010, as well as bibliographies of selected studies. We sought original studies that reported quantitative or qualitative data about medical students' exposure to pharmaceutical marketing, their attitudes about marketing practices, relationships with industry, and related pharmaceutical policy issues. Studies were separated, where possible, into those that addressed preclinical versus clinical training, and were quality rated using a standard methodology. Thirty two studies met inclusion criteria. We found that 40%-100% of medical students reported interacting with the pharmaceutical industry. A substantial proportion of students (13%-69%) were reported as believing that gifts from industry influence prescribing. Eight studies reported a correlation between frequency of contact and favorable attitudes toward industry interactions. Students were more approving of gifts to physicians or medical students than to government officials. Certain attitudes appeared to change during medical school, though a time trend was not performed; for example, clinical students (53%-71%) were more likely than preclinical students (29%-62%) to report that promotional information helps educate about new drugs. CONCLUSIONS: Undergraduate medical education provides substantial contact with pharmaceutical marketing, and the extent of such contact is associated with positive attitudes about marketing and skepticism about negative implications of these interactions. These results support future research into the association between exposure and attitudes, as well as any modifiable factors that contribute to attitudinal changes during medical education. Please see later in the article for the Editors' Summary. PMID- 21629686 TI - Short-term memory maintenance of object locations during active navigation: which working memory subsystem is essential? AB - The goal of the present study was to examine the extent to which working memory supports the maintenance of object locations during active spatial navigation. Participants were required to navigate a virtual environment and to encode the location of a target object. In the subsequent maintenance period they performed one of three secondary tasks that were designed to selectively load visual, verbal or spatial working memory subsystems. Thereafter participants re-entered the environment and navigated back to the remembered location of the target. We found that while navigation performance in participants with high navigational ability was impaired only by the spatial secondary task, navigation performance in participants with poor navigational ability was impaired equally by spatial and verbal secondary tasks. The visual secondary task had no effect on navigation performance. Our results extend current knowledge by showing that the differential engagement of working memory subsystems is determined by navigational ability. PMID- 21629687 TI - An Algebro-topological description of protein domain structure. AB - The space of possible protein structures appears vast and continuous, and the relationship between primary, secondary and tertiary structure levels is complex. Protein structure comparison and classification is therefore a difficult but important task since structure is a determinant for molecular interaction and function. We introduce a novel mathematical abstraction based on geometric topology to describe protein domain structure. Using the locations of the backbone atoms and the hydrogen bonds, we build a combinatorial object--a so called fatgraph. The description is discrete yet gives rise to a 2-dimensional mathematical surface. Thus, each protein domain corresponds to a particular mathematical surface with characteristic topological invariants, such as the genus (number of holes) and the number of boundary components. Both invariants are global fatgraph features reflecting the interconnectivity of the domain by hydrogen bonds. We introduce the notion of robust variables, that is variables that are robust towards minor changes in the structure/fatgraph, and show that the genus and the number of boundary components are robust. Further, we investigate the distribution of different fatgraph variables and show how only four variables are capable of distinguishing different folds. We use local (secondary) and global (tertiary) fatgraph features to describe domain structures and illustrate that they are useful for classification of domains in CATH. In addition, we combine our method with two other methods thereby using primary, secondary, and tertiary structure information, and show that we can identify a large percentage of new and unclassified structures in CATH. PMID- 21629688 TI - Elevated CO2 influences nematode-induced defense responses of tomato genotypes differing in the JA pathway. AB - Rising atmospheric CO(2) concentrations can affect the induced defense of plants against chewing herbivores but little is known about whether elevated CO(2) can change the induced defense of plants against parasitic nematodes. This study examined the interactions between the root-knot nematode Meloidogyne incognita and three isogenic tomato (Lycopersicon esculentum) genotypes grown under ambient (390 ppm) and elevated (750 ppm) CO(2) in growth chambers. In a previous study with open-top chambers in the field, we reported that elevated CO(2) increased the number of nematode-induced root galls in a JA-defense-dominated genotype but not in a wild-type or JA-defense-recessive genotype. In the current study, we tested the hypothesis that elevated CO(2) will favor the salicylic acid (SA) pathway defense but repress the jasmonic acid (JA)-pathway defense of plants against plant-parasitic nematodes. Our data showed that elevated CO(2) reduced the JA-pathway defense against M. incognita in the wild-type and in a genotype in which defense is dominated by the JA pathway (a JA-defense-dominated genotype) but up-regulated the SA-pathway defense in the wild type and in a JA-defense recessive genotype (jasmonate-deficient mutant). Our results suggest that, in terms of defense genes, secondary metabolites, and volatile organic compounds, induced defense of nematode-infected plants could be affected by elevated CO(2), and that CO(2)-induced changes of plant resistance may lead to genotype-specific responses of plants to nematodes under elevated CO(2). The changes in resistance against nematodes, however, were small relative to those reported for chewing insects. PMID- 21629689 TI - cDNA sequence and Fab crystal structure of HL4E10, a hamster IgG lambda light chain antibody stimulatory for gammadelta T cells. AB - Hamsters are widely used to generate monoclonal antibodies against mouse, rat, and human antigens, but sequence and structural information for hamster immunoglobulins is sparse. To our knowledge, only three hamster IgG sequences have been published, all of which use kappa light chains, and no three dimensional structure of a hamster antibody has been reported. We generated antibody HL4E10 as a probe to identify novel costimulatory molecules on the surface of gammadelta T cells which lack the traditional alphabeta T cell co receptors CD4, CD8, and the costimulatory molecule CD28. HL4E10 binding to gammadelta T cell, surface-expressed, Junctional Adhesion Molecule-Like (JAML) protein leads to potent costimulation via activation of MAP kinase pathways and cytokine production, resulting in cell proliferation. The cDNA sequence of HL4E10 is the first example of a hamster lambda light chain and only the second known complete hamster heavy chain sequence. The crystal structure of the HL4E10 Fab at 2.95 A resolution reveals a rigid combining site with pockets faceted by solvent exposed tyrosine residues, which are structurally optimized for JAML binding. The characterization of HL4E10 thus comprises a valuable addition to the spartan database of hamster immunoglobulin genes and structures. As the HL4E10 antibody is uniquely costimulatory for gammadelta T cells, humanized versions thereof may be of clinical relevance in treating gammadelta T cell dysfunction-associated diseases, such as chronic non-healing wounds and cancer. PMID- 21629690 TI - Perceived positions determine crowding. AB - Crowding is a fundamental bottleneck in object recognition. In crowding, an object in the periphery becomes unrecognizable when surrounded by clutter or distractor objects. Crowding depends on the positions of target and distractors, both their eccentricity and their relative spacing. In all previous studies, position has been expressed in terms of retinal position. However, in a number of situations retinal and perceived positions can be dissociated. Does retinal or perceived position determine the magnitude of crowding? Here observers performed an orientation judgment on a target Gabor patch surrounded by distractors that drifted toward or away from the target, causing an illusory motion-induced position shift. Distractors in identical physical positions led to worse performance when they drifted towards the target (appearing closer) versus away from the target (appearing further). This difference in crowding corresponded to the difference in perceived positions. Further, the perceptual mislocalization was necessary for the change in crowding, and both the mislocalization and crowding scaled with drift speed. The results show that crowding occurs after perceived positions have been assigned by the visual system. Crowding does not operate in a purely retinal coordinate system; perceived positions need to be taken into account. PMID- 21629691 TI - Poor regenerative outcome after skeletal muscle necrosis induced by Bothrops asper venom: alterations in microvasculature and nerves. AB - BACKGROUND: Viperid snakebite envenoming is characterized by prominent local tissue damage, including muscle necrosis. A frequent outcome of such local pathology is deficient skeletal muscle regeneration, which causes muscle dysfunction, muscle loss and fibrosis, thus provoking permanent sequelae that greatly affect the quality of life of patients. The causes of such poor regenerative outcome of skeletal muscle after viperid snakebites are not fully understood. METHODOLOGY/PRINCIPAL FINDINGS: A murine model of muscle necrosis and regeneration was adapted to study the effects of the venom and isolated toxins of Bothrops asper, the medically most important snake in Central America. Gastrocnemius muscle was injected with either B. asper venom, a myotoxic phospholipase A(2) (Mtx), a hemorrhagic metalloproteinase (SVMP), or saline solution. At various time intervals, during one month, tissue samples were collected and analyzed by histology, and by immunocytochemical and immunohistochemical techniques aimed at detecting muscle fibers, collagen, endothelial cells, myoblasts, myotubes, macrophages, TUNEL-positive nuclei, and axons. A successful regenerative response was observed in muscle injected with Mtx, which induces myonecrosis but does not affect the microvasculature. In contrast, poor regeneration, with fibrosis and atrophic fibers, occurred when muscle was injected with venom or SVMP, both of which provoke necrosis, microvascular damage leading to hemorrhage, and poor axonal regeneration. CONCLUSIONS/SIGNIFICANCE: The deficient skeletal muscle regeneration after injection of B. asper venom is likely to depend on the widespread damage to the microvasculature, which affects the removal of necrotic debris by phagocytes, and the provision of nutrients and oxygen required for regeneration. In addition, deficient axonal regeneration is likely to contribute to the poor regenerative outcome in this model. PMID- 21629692 TI - Suppression of estrogen receptor transcriptional activity by connective tissue growth factor. AB - Secreted growth factors have been shown to stimulate the transcriptional activity of estrogen receptors (ER) that are responsible for many biological processes. However, whether these growth factors physically interact with ER remains unclear. Here, we show for the first time that connective tissue growth factor (CTGF) physically and functionally associates with ER. CTGF interacted with ER both in vitro and in vivo. CTGF interacted with ER DNA-binding domain. ER interaction region in CTGF was mapped to the thrombospondin type I repeat, a cell attachment motif. Overexpression of CTGF inhibited ER transcriptional activity as well as the expression of estrogen-responsive genes, including pS2 and cathepsin D. Reduction of endogenous CTGF with CTGF small interfering RNA enhanced ER transcriptional activity. The interaction between CTGF and ER is required for the repression of estrogen-responsive transcription by CTGF. Moreover, CTGF reduced ER protein expression, whereas the CTGF mutant that did not repress ER transcriptional activity also did not alter ER protein levels. The results suggested the transcriptional regulation of estrogen signaling through interaction between CTGF and ER, and thus may provide a novel mechanism by which cross-talk between secreted growth factor and ER signaling pathways occurs. PMID- 21629693 TI - Nucleolar localization of RNA binding proteins induced by actinomycin D and heat shock in Trypanosoma cruzi. AB - In this work we show that under Actinomycin D (ActD) treatment, several RNA Binding Proteins (RBPs) involved in mRNA metabolism are relocalized into the nucleolus in Trypanosoma cruzi as a specific stress response. ATP depletion as well as kinase inhibition markedly reduced the nucleolar localization response, suggesting that an energy-dependent transport modulated by the phosphorylation status of the parasite might be required. Deletion analyses in one of such proteins, TcSR62, showed that a domain bearing basic amino acids located in the COOH terminal region was sufficient to promote its nucleolar relocalization. Interestingly, we showed that in addition to RBPs, poly(A)+ RNA is also accumulated into the nucleolus in response to ActD treatment. Finally, we found out that nucleolar relocalization of RBPs is also triggered by severe heat shock in a reversible way. Together, these results suggest that the nucleolus of an early divergent eukaryote is either able to sequester key factors related to mRNA metabolism in response to transcriptional stress or behaves as a RBP processing center, arguing in favour to the hypothesis that the non-traditional features of the nucleolus could be acquired early during evolution. PMID- 21629694 TI - On the adaptive partition approach to the detection of multiple change-points. AB - With an adaptive partition procedure, we can partition a "time course" into consecutive non-overlapped intervals such that the population means/proportions of the observations in two adjacent intervals are significantly different at a given level . However, the widely used recursive combination or partition procedures do not guarantee a global optimization. We propose a modified dynamic programming algorithm to achieve a global optimization. Our method can provide consistent estimation results. In a comprehensive simulation study, our method shows an improved performance when it is compared to the recursive combination/partition procedures. In practice, can be determined based on a cross validation procedure. As an application, we consider the well-known Pima Indian Diabetes data. We explore the relationship among the diabetes risk and several important variables including the plasma glucose concentration, body mass index and age. PMID- 21629695 TI - Evolutionary relationships among Chlamydophila abortus variant strains inferred by rRNA secondary structure-based phylogeny. AB - The evolutionary relationships among known Chlamydophila abortus variant strains including the LLG and POS, previously identified as being highly distinct, were investigated based on rRNA secondary structure information. PCR-amplified overlapping fragments of the 16S, 16S-23S intergenic spacer (IS), and 23S domain I rRNAs were subjected to cloning and sequencing. Secondary structure analysis revealed the presence of transitional single nucleotide variations (SNVs), two of which occurred in loops, while seven in stem regions that did not result in compensatory substitutions. Notably, only two SNVs, in 16S and 23S, occurred within evolutionary variable regions. Maximum likelihood and Bayesian phylogeny reconstructions revealed that C. abortus strains could be regarded as representing two distinct lineages, one including the "classical" C. abortus strains and the other the "LLG/POS variant", with the type strain B577(T) possibly representing an intermediate of the two lineages. The two C. abortus lineages shared three unique (apomorphic) characters in the 23S domain I and 16S 23S IS, but interestingly lacked synapomorphies in the 16S rRNA. The two lineages could be distinguished on the basis of eight positions; four of these comprised residues that appeared to be signature or unique for the "classical" lineage, while three were unique for the "LLG/POS variant". The U277 (E. coli numbering) signature character, corresponding to a highly conserved residue of the 16S molecule, and the unique G681 residue, conserved in a functionally strategic region also of 16S, are the most pronounced attributes (autapomorphies) of the "classical" and the "LLG/POS variant" lineages, respectively. Both lineages were found to be descendants of a common ancestor with the Prk/Daruma C. psittaci variant. Compared with the "classical", the "LLG/POS variant" lineage has retained more ancestral features. The current rRNA secondary structure-based analysis and phylogenetic inference reveal new insights into how these two C. abortus lineages have differentiated during their evolution. PMID- 21629696 TI - Variation in the analysis of positively selected sites using nonsynonymous/synonymous rate ratios: an example using influenza virus. AB - Sites in a gene showing the nonsynonymous/synonymous rate ratio (omega) >1 have been frequently identified to be under positive selection. To examine the performance of such analysis, sites of the omega ratio >1 in the HA1 gene of H3N2 subtype human influenza viruses were identified from seven overlapping sequence data sets in this study. Our results showed that the sites of the omega ratio >1 were of significant variation among the data sets even though they targeted similar clusters, indicating that the analysis is likely to be either of low sensitivity or of low specificity in identifying sites under positive selection. Most (43/45) of the sites showing omega >1 calculated from at least one data set are involved in B-cell epitopes which cover less than a half sites in the protein, suggesting that the analysis is likely to be of low sensitivity rather than of low specificity. It was further found that the analysis sensitivity could not be enhanced by including more sequences or covering longer time intervals. Previously some reports also likely identified only a portion of the sites under positive selection in the viral gene using the omega ratio. Low sensitivity of the analysis may result from that some sites under positive selection in the gene are also under negative (purifying) selection simultaneously for functional constrains, and so their omega ratios could be <1. Theoretically, the sites under the two opposite selection forces at the same time favor only certain nonsynonymous changes, e.g. those changing the antigenicity of the gene and maintaining the gene function. This study also suggested that sometimes we can identify more sites under positive selection using the omega ratio by integrating the positively selected sites estimated from multiple data sets. PMID- 21629697 TI - A high-throughput platform for lentiviral overexpression screening of the human ORFeome. AB - In response to the growing need for functional analysis of the human genome, we have developed a platform for high-throughput functional screening of genes overexpressed from lentiviral vectors. Protein-coding human open reading frames (ORFs) from the Mammalian Gene Collection were transferred into lentiviral expression vector using the highly efficient Gateway recombination cloning. Target ORFs were inserted into the vector downstream of a constitutive promoter and upstream of an IRES controlled GFP reporter, so that their transfection, transduction and expression could be monitored by fluorescence. The expression plasmids and viral packaging plasmids were combined and transfected into 293T cells to produce virus, which was then used to transduce the screening cell line. We have optimised the transfection and transduction procedures so that they can be performed using robotic liquid handling systems in arrayed 96-well microplate, one-gene-per-well format, without the need to concentrate the viral supernatant. Since lentiviruses can infect both dividing and non-dividing cells, this system can be used to overexpress human ORFs in a broad spectrum of experimental contexts. We tested the platform in a 1990 gene pilot screen for genes that can increase proliferation of the non-tumorigenic mammary epithelial cell line MCF 10A after removal of growth factors. Transduced cells were labelled with the nucleoside analogue 5-ethynyl-2'-deoxyuridine (EdU) to detect cells progressing through S phase. Hits were identified using high-content imaging and statistical analysis and confirmed with vectors using two different promoters (CMV and EF1alpha). The screen demonstrates the reliability, versatility and utility of our screening platform, and identifies novel cell cycle/proliferative activities for a number of genes. PMID- 21629699 TI - Preclinical development of an in vivo BCG challenge model for testing candidate TB vaccine efficacy. AB - There is an urgent need for an immunological correlate of protection against tuberculosis (TB) with which to evaluate candidate TB vaccines in clinical trials. Development of a human challenge model of Mycobacterium tuberculosis (M.tb) could facilitate the detection of such correlate(s). Here we propose a novel in vivo Bacille Calmette-Guerin (BCG) challenge model using BCG immunization as a surrogate for M.tb infection. Culture and quantitative PCR methods have been developed to quantify BCG in the skin, using the mouse ear as a surrogate for human skin. Candidate TB vaccines have been evaluated for their ability to protect against a BCG skin challenge, using this model, and the results indicate that protection against a BCG skin challenge is predictive of BCG vaccine efficacy against aerosol M.tb challenge. Translation of these findings to a human BCG challenge model could enable more rapid assessment and down selection of candidate TB vaccines and ultimately the identification of an immune correlate of protection. PMID- 21629698 TI - Gene expression profiling and association with prion-related lesions in the medulla oblongata of symptomatic natural scrapie animals. AB - The pathogenesis of natural scrapie and other prion diseases remains unclear. Examining transcriptome variations in infected versus control animals may highlight new genes potentially involved in some of the molecular mechanisms of prion-induced pathology. The aim of this work was to identify disease-associated alterations in the gene expression profiles of the caudal medulla oblongata (MO) in sheep presenting the symptomatic phase of natural scrapie. The gene expression patterns in the MO from 7 sheep that had been naturally infected with scrapie were compared with 6 controls using a Central Veterinary Institute (CVI) custom designed 4*44K microarray. The microarray consisted of a probe set on the previously sequenced ovine tissue library by CVI and was supplemented with all of the Ovis aries transcripts that are currently publicly available. Over 350 probe sets displayed greater than 2-fold changes in expression. We identified 148 genes from these probes, many of which encode proteins that are involved in the immune response, ion transport, cell adhesion, and transcription. Our results confirm previously published gene expression changes that were observed in murine models with induced scrapie. Moreover, we have identified new genes that exhibit differential expression in scrapie and could be involved in prion neuropathology. Finally, we have investigated the relationship between gene expression profiles and the appearance of the main scrapie-related lesions, including prion protein deposition, gliosis and spongiosis. In this context, the potential impacts of these gene expression changes in the MO on scrapie development are discussed. PMID- 21629700 TI - Production of multiple brain-like ganglioside species is dispensable for fas induced apoptosis of lymphoid cells. AB - Activation of an acid sphingomyelinase (aSMase) leading to a biosynthesis of GD3 disialoganglioside has been associated with Fas-induced apoptosis of lymphoid cells. The present study was undertaken to clarify the role of this enzyme in the generation of gangliosides during apoptosis triggered by Fas ligation. The issue was addressed by using aSMase-deficient and aSMase-corrected cell lines derived from Niemann-Pick disease (NPD) patients. Fas cross-linking elicited a rapid production of large amounts of complex a- and b-series species of gangliosides with a pattern and a chromatographic behavior as single bands reminiscent of brain gangliosides. The gangliosides were synthesized within the first ten minutes and completely disappeared within thirty minutes after stimulation. Noteworthy is the observation that GD3 was not the only ganglioside produced. The production of gangliosides and the onset of apoptotic hallmarks occurred similarly in both aSMase-deficient and aSMase-corrected NPD lymphoid cells, indicating that aSMase activation is not accountable for ganglioside generation. Hampering ganglioside production by inhibiting the key enzyme glucosylceramide synthase did not abrogate the apoptotic process. In addition, GM3 synthase deficient lymphoid cells underwent Fas-induced apoptosis, suggesting that gangliosides are unlikely to play an indispensable role in transducing Fas induced apoptosis of lymphoid cells. PMID- 21629701 TI - Spliced leader RNAs, mitochondrial gene frameshifts and multi-protein phylogeny expand support for the genus Perkinsus as a unique group of alveolates. AB - The genus Perkinsus occupies a precarious phylogenetic position. To gain a better understanding of the relationship between perkinsids, dinoflagellates and other alveolates, we analyzed the nuclear-encoded spliced-leader (SL) RNA and mitochondrial genes, intron prevalence, and multi-protein phylogenies. In contrast to the canonical 22-nt SL found in dinoflagellates (DinoSL), P. marinus has a shorter (21-nt) and a longer (22-nt) SL with slightly different sequences than DinoSL. The major SL RNA transcripts range in size between 80-83 nt in P. marinus, and ~ 83 nt in P. chesapeaki, significantly larger than the typical <= 56-nt dinoflagellate SL RNA. In most of the phylogenetic trees based on 41 predicted protein sequences, P. marinus branched at the base of the dinoflagellate clade that included the ancient taxa Oxyrrhis and Amoebophrya, sister to the clade of apicomplexans, and in some cases clustered with apicomplexans as a sister to the dinoflagellate clade. Of 104 Perkinsus spp. genes examined 69.2% had introns, a higher intron prevalence than in dinoflagellates. Examination of Perkinsus spp. mitochondrial cytochrome B and cytochrome C oxidase subunit I genes and their cDNAs revealed no mRNA editing, but these transcripts can only be translated when frameshifts are introduced at every AGG and CCC codon as if AGGY codes for glycine and CCCCU for proline. These results, along with the presence of the numerous uncharacterized 'marine alveolate group I' and Perkinsus-like lineages separating perkinsids from core dinoflagellates, expand support for the affiliation of the genus Perkinsus with an independent lineage (Perkinsozoa) positioned between the phyla of Apicomplexa and Dinoflagellata. PMID- 21629702 TI - Adaptive movement compensation for in vivo imaging of fast cellular dynamics within a moving tissue. AB - In vivo non-linear optical microscopy has been essential to advance our knowledge of how intact biological systems work. It has been particularly enabling to decipher fast spatiotemporal cellular dynamics in neural networks. The power of the technique stems from its optical sectioning capability that in turn also limits its application to essentially immobile tissue. Only tissue not affected by movement or in which movement can be physically constrained can be imaged fast enough to conduct functional studies at high temporal resolution. Here, we show dynamic two-photon Ca(2+) imaging in the spinal cord of a living rat at millisecond time scale, free of motion artifacts using an optical stabilization system. We describe a fast, non-contact adaptive movement compensation approach, applicable to rough and weakly reflective surfaces, allowing real-time functional imaging from intrinsically moving tissue in live animals. The strategy involves enslaving the position of the microscope objective to that of the tissue surface in real-time through optical monitoring and a closed feedback loop. The performance of the system allows for efficient image locking even in conditions of random or irregular movements. PMID- 21629703 TI - Structural heterogeneity and quantitative FRET efficiency distributions of polyprolines through a hybrid atomistic simulation and Monte Carlo approach. AB - Forster Resonance Energy Transfer (FRET) experiments probe molecular distances via distance dependent energy transfer from an excited donor dye to an acceptor dye. Single molecule experiments not only probe average distances, but also distance distributions or even fluctuations, and thus provide a powerful tool to study biomolecular structure and dynamics. However, the measured energy transfer efficiency depends not only on the distance between the dyes, but also on their mutual orientation, which is typically inaccessible to experiments. Thus, assumptions on the orientation distributions and averages are usually made, limiting the accuracy of the distance distributions extracted from FRET experiments. Here, we demonstrate that by combining single molecule FRET experiments with the mutual dye orientation statistics obtained from Molecular Dynamics (MD) simulations, improved estimates of distances and distributions are obtained. From the simulated time-dependent mutual orientations, FRET efficiencies are calculated and the full statistics of individual photon absorption, energy transfer, and photon emission events is obtained from subsequent Monte Carlo (MC) simulations of the FRET kinetics. All recorded emission events are collected to bursts from which efficiency distributions are calculated in close resemblance to the actual FRET experiment, taking shot noise fully into account. Using polyproline chains with attached Alexa 488 and Alexa 594 dyes as a test system, we demonstrate the feasibility of this approach by direct comparison to experimental data. We identified cis-isomers and different static local environments as sources of the experimentally observed heterogeneity. Reconstructions of distance distributions from experimental data at different levels of theory demonstrate how the respective underlying assumptions and approximations affect the obtained accuracy. Our results show that dye fluctuations obtained from MD simulations, combined with MC single photon kinetics, provide a versatile tool to improve the accuracy of distance distributions that can be extracted from measured single molecule FRET efficiencies. PMID- 21629704 TI - Autophagic and apoptotic effects of HDAC inhibitors on cancer cells. AB - Because epigenetic alterations are believed to be involved in the repression of tumor suppressor genes and the promotion of tumorigenesis in cancers, novel compounds endowed with histone deacetylase (HDAC) inhibitory activity are an attractive therapeutic approach. Indeed, the potential of HDAC inhibitors for cancer therapy has been explored in preclinical models, and some agents approved for hematologic malignancies have reached the clinical setting. HDAC inhibitors are able to mediate the induction of both apoptosis and autophagy, which are related to anticancer activity in a variety of cancer cell lines. Given the inherent resistance to apoptosis that characterizes cancer, the targeting of alternative pathways is an attractive strategy to improve anti-tumor therapy. The activation of autophagy represents novel cancer treatment targets. This paper aims to critically discuss how the anticancer potential of HDAC inhibitors may elicit a response to human cancers through different cell pathways leading to cell death. PMID- 21629705 TI - Mitochondria and PGC-1alpha in Aging and Age-Associated Diseases. AB - Aging is the most significant risk factor for a range of degenerative disease such as cardiovascular, neurodegenerative and metabolic disorders. While the cause of aging and its associated diseases is multifactorial, mitochondrial dysfunction has been implicated in the aging process and the onset and progression of age-associated disorders. Recent studies indicate that maintenance of mitochondrial function is beneficial in the prevention or delay of age associated diseases. A central molecule seems to be the peroxisome proliferator activated receptor gamma coactivator alpha (PGC-1alpha), which is the key regulator of mitochondrial biogenesis. Besides regulating mitochondrial function, PGC-1alpha targets several other cellular processes and thereby influences cell fate on multiple levels. This paper discusses how mitochondrial function and PGC 1alpha are affected in age-associated diseases and how modulation of PGC-1alpha might offer a therapeutic potential for age-related pathology. PMID- 21629706 TI - Metals and Alzheimer's disease. PMID- 21629707 TI - The role of natural killer cells in sepsis. AB - Severe sepsis and septic shock are still deadly conditions urging to develop novel therapies. A better understanding of the complex modifications of the immune system of septic patients is needed for the development of innovative immunointerventions. Natural killer (NK) cells are characterized as CD3( )NKp46(+)CD56(+) cells that can be cytotoxic and/or produce high amounts of cytokines such as IFN-gamma. NK cells are also engaged in crosstalks with other immune cells, such as dendritic cells, macrophages, and neutrophils. During the early stage of septic shock, NK cells may play a key role in the promotion of the systemic inflammation, as suggested in mice models. Alternatively, at a later stage, NK cells-acquired dysfunction could favor nosocomial infections and mortality. Standardized biological tools defining patients' NK cell status during the different stages of sepsis are mandatory to guide potential immuno interventions. Herein, we review the potential role of NK cells during severe sepsis and septic shock. PMID- 21629708 TI - Lateral Spread of Orientation Selectivity in V1 is Controlled by Intracortical Cooperativity. AB - Neurons in the primary visual cortex receive subliminal information originating from the periphery of their receptive fields (RF) through a variety of cortical connections. In the cat primary visual cortex, long-range horizontal axons have been reported to preferentially bind to distant columns of similar orientation preferences, whereas feedback connections from higher visual areas provide a more diverse functional input. To understand the role of these lateral interactions, it is crucial to characterize their effective functional connectivity and tuning properties. However, the overall functional impact of cortical lateral connections, whatever their anatomical origin, is unknown since it has never been directly characterized. Using direct measurements of postsynaptic integration in cat areas 17 and 18, we performed multi-scale assessments of the functional impact of visually driven lateral networks. Voltage-sensitive dye imaging showed that local oriented stimuli evoke an orientation-selective activity that remains confined to the cortical feedforward imprint of the stimulus. Beyond a distance of one hypercolumn, the lateral spread of cortical activity gradually lost its orientation preference approximated as an exponential with a space constant of about 1 mm. Intracellular recordings showed that this loss of orientation selectivity arises from the diversity of converging synaptic input patterns originating from outside the classical RF. In contrast, when the stimulus size was increased, we observed orientation-selective spread of activation beyond the feedforward imprint. We conclude that stimulus-induced cooperativity enhances the long-range orientation-selective spread. PMID- 21629709 TI - Multimodal Imaging of Alzheimer Pathophysiology in the Brain's Default Mode Network. AB - The spatial correlations between the brain's default mode network (DMN) and the brain regions known to develop pathophysiology in Alzheimer's disease (AD) have recently attracted much attention. In this paper, we compare results of different functional and structural imaging modalities, including MRI and PET, and highlight different patterns of anomalies observed within the DMN. Multitracer PET imaging in subjects with and without dementia has demonstrated that [C-11]PIB and [F-18]FDDNP-binding patterns in patients with AD overlap within nodes of the brain's default network including the prefrontal, lateral parietal, lateral temporal, and posterior cingulate cortices, with the exception of the medial temporal cortex (especially, the hippocampus) where significant discrepancy between increased [F-18]FDDNP binding and negligible [C-11]PIB-binding was observed. [F-18]FDDNP binding in the medial temporal cortex-a key constituent of the DMN-coincides with both the presence of amyloid and tau pathology, and also with cortical areas with maximal atrophy as demonstrated by T1-weighted MR imaging of AD patients. PMID- 21629710 TI - Morphofunctional and Biochemical Approaches for Studying Mitochondrial Changes during Myoblasts Differentiation. AB - This study describes mitochondrial behaviour during the C2C12 myoblast differentiation program and proposes a proteomic approach to mitochondria integrated with classical morphofunctional and biochemical analyses. Mitochondrial ultrastructure variations were determined by transmission electron microscopy; mitochondrial mass and membrane potential were analysed by Mitotracker Green and JC-1 stains and by epifluorescence microscope. Expression of PGC1alpha, NRF1alpha, and Tfam genes controlling mitochondrial biogenesis was studied by real-time PCR. The mitochondrial functionality was tested by cytochrome c oxidase activity and COXII expression. Mitochondrial proteomic profile was also performed. These assays showed that mitochondrial biogenesis and activity significantly increase in differentiating myotubes. The proteomic profile identifies 32 differentially expressed proteins, mostly involved in oxidative metabolism, typical of myotubes formation. Other notable proteins, such as superoxide dismutase (MnSOD), a cell protection molecule, and voltage dependent anion-selective channel protein (VDAC1) involved in the mitochondria mediated apoptosis, were found to be regulated by the myogenic process. The integration of these approaches represents a helpful tool for studying mitochondrial dynamics, biogenesis, and functionality in comparative surveys on mitochondrial pathogenic or senescent satellite cells. PMID- 21629711 TI - Does EEG montage influence Alzheimer's disease electroclinic diagnosis? AB - There is not a specific Alzheimer's disease (AD) diagnostic test. AD diagnosis relies on clinical history, neuropsychological, and laboratory tests, neuroimaging and electroencephalography. Therefore, new approaches are necessary to enable earlier and more accurate diagnosis and to measure treatment results. Quantitative EEG (qEEG) can be used as a diagnostic tool in selected cases. The aim of this study was to answer if distinct electrode montages have different sensitivity when differentiating controls from AD patients. We analyzed EEG spectral peaks (delta, theta, alpha, beta, and gamma bands), and we compared references (Biauricular, Longitudinal bipolar, Crossed bipolar, Counterpart bipolar, and Cz reference). Support Vector Machines and Logistic Regression classifiers showed Counterpart bipolar montage as the most sensitive electrode combination. Our results suggest that Counterpart bipolar montage is the best choice to study EEG spectral peaks of controls versus AD. PMID- 21629712 TI - Older adults' perceptions of clinical fall prevention programs: a qualitative study. AB - Objective. To investigate motivational factors and barriers to participating in fall risk assessment and management programs among diverse, low-income, community dwelling older adults who had experienced a fall. Methods. Face-to-face interviews with 20 elderly who had accepted and 19 who had not accepted an invitation to an assessment by one of two fall prevention programs. Interviews covered healthy aging, core values, attributions/consequences of the fall, and barriers/benefits of fall prevention strategies and programs. Results. Joiners and nonjoiners of fall prevention programs were similar in their experience of loss associated with aging, core values they expressed, and emotional response to falling. One difference was that those who participated endorsed that they "needed" the program, while those who did not participate expressed a lack of need. Conclusions. Interventions targeted at a high-risk group need to address individual beliefs as well as structural and social factors (transportation issues, social networks) to enhance participation. PMID- 21629713 TI - Regulation of cell survival mechanisms in Alzheimer's disease by glycogen synthase kinase-3. AB - A pivotal role has emerged for glycogen synthase kinase-3 (GSK3) as an important contributor to Alzheimer's disease pathology. Evidence for the involvement of GSK3 in Alzheimer's disease pathology and neuronal loss comes from studies of GSK3 overexpression, GSK3 localization studies, multiple relationships between GSK3 and amyloid beta-peptide (Abeta), interactions between GSK3 and the microtubule-associated tau protein, and GSK3-mediated apoptotic cell death. Apoptotic signaling proceeds by either an intrinsic pathway or an extrinsic pathway. GSK3 is well established to promote intrinsic apoptotic signaling induced by many insults, several of which may contribute to neuronal loss in Alzheimer's disease. Particularly important is evidence that GSK3 promotes intrinsic apoptotic signaling induced by Abeta. GSK3 appears to promote intrinsic apoptotic signaling by modulating proteins in the apoptosis signaling pathway and by modulating transcription factors that regulate the expression of proteins involved in apoptosis. Thus, GSK3 appears to contribute to several neuropathological mechanisms in Alzheimer's disease, including apoptosis-mediated neuronal loss. PMID- 21629714 TI - Electroencephalographic rhythms in Alzheimer's disease. AB - Physiological brain aging is characterized by synapses loss and neurodegeneration that slowly lead to an age-related decline of cognition. Neural/synaptic redundancy and plastic remodelling of brain networking, also due to mental and physical training, promotes maintenance of brain activity in healthy elderly subjects for everyday life and good social behaviour and intellectual capabilities. However, age is the major risk factor for most common neurodegenerative disorders that impact on cognition, like Alzheimer's disease (AD). Brain electromagnetic activity is a feature of neuronal network function in various brain regions. Modern neurophysiological techniques, such as electroencephalography (EEG) and event-related potentials (ERPs), are useful tools in the investigation of brain cognitive function in normal and pathological aging with an excellent time resolution. These techniques can index normal and abnormal brain aging analysis of corticocortical connectivity and neuronal synchronization of rhythmic oscillations at various frequencies. The present review suggests that discrimination between physiological and pathological brain aging clearly emerges at the group level, with suggested applications also at the level of single individual. The possibility of combining the use of EEG together with biological/neuropsychological markers and structural/functional imaging is promising for a low-cost, non-invasive, and widely available assessment of groups of individuals at-risk. PMID- 21629715 TI - A biophysical model of cell adhesion mediated by immunoadhesin drugs and antibodies. AB - A promising direction in drug development is to exploit the ability of natural killer cells to kill antibody-labeled target cells. Monoclonal antibodies and drugs designed to elicit this effect typically bind cell-surface epitopes that are overexpressed on target cells but also present on other cells. Thus it is important to understand adhesion of cells by antibodies and similar molecules. We present an equilibrium model of such adhesion, incorporating heterogeneity in target cell epitope density, nonspecific adhesion forces, and epitope immobility. We compare with experiments on the adhesion of Jurkat T cells to bilayers containing the relevant natural killer cell receptor, with adhesion mediated by the drug alefacept. We show that a model in which all target cell epitopes are mobile and available is inconsistent with the data, suggesting that more complex mechanisms are at work. We hypothesize that the immobile epitope fraction may change with cell adhesion, and we find that such a model is more consistent with the data, although discrepancies remain. We also quantitatively describe the parameter space in which binding occurs. Our model elaborates substantially on previous work, and our results offer guidance for the refinement of therapeutic immunoadhesins. Furthermore, our comparison with data from Jurkat T cells also points toward mechanisms relating epitope immobility to cell adhesion. PMID- 21629716 TI - Targeting Glycogen Synthase Kinase-3beta for Therapeutic Benefit against Oxidative Stress in Alzheimer's Disease: Involvement of the Nrf2-ARE Pathway. AB - Specific regions of the Alzheimer's disease (AD) brain are burdened with extracellular protein deposits, the accumulation of which is concomitant with a complex cascade of overlapping events. Many of these pathological processes produce oxidative stress. Under normal conditions, oxidative stress leads to the activation of defensive gene expression that promotes cell survival. At the forefront of defence is the nuclear factor erythroid 2-related factor 2 (Nrf2), a transcription factor that regulates a broad spectrum of protective genes. Glycogen synthase kinase-3beta (GSK-3beta) regulates Nrf2, thus making this kinase a potential target for therapeutic intervention aiming to boost the protective activation of Nrf2. This paper aims to review the neuroprotective role of Nrf2 in AD, with special emphasis on the role of GSK-3beta in the regulation of the Nrf2 pathway. We also examine the potential of inducing GSK-3beta by small molecule activators, dithiocarbamates, which potentially exert their beneficial therapeutic effects via the activation of the Nrf2 pathway. PMID- 21629717 TI - Dysregulated Neuronal Activity Patterns Implicate Corticostriatal Circuit Dysfunction in Multiple Rodent Models of Huntington's Disease. AB - Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder that targets the corticostriatal system and results in progressive deterioration of cognitive, emotional, and motor skills. Although cortical and striatal neurons are widely studied in animal models of HD, there is little information on neuronal function during expression of the HD behavioral phenotype. To address this knowledge gap, we used chronically implanted micro-wire bundles to record extracellular spikes and local field potentials (LFPs) in truncated (R6/1 and R6/2) and full-length (knock-in, KI) mouse models as well as in transgenic HD rats (tgHD rats) behaving in an open-field arena. Spike activity was recorded in the striatum of all models and in prefrontal cortex (PFC) of R6/2 and KI mice, and in primary motor cortex (M1) of R6/2 mice. We also recorded LFP activity in R6/2 striatum. All HD models exhibited altered neuronal activity relative to wild type (WT) controls. Although there was no consistent effect on firing rate across models and brain areas, burst firing was reduced in striatum, PFC, and M1 of R6/2 mice, and in striatum of KI mice. Consistent with a decline in bursting, the inter-spike-interval coefficient of variation was reduced in all regions of all models, except PFC of KI mice and striatum of tgHD rats. Among simultaneously recorded neuron pairs, correlated firing was reduced in all brain regions of all models, while coincident bursting, which measures the temporal overlap between bursting pairs, was reduced in striatum of all models as well as in M1 of R6/2s. Preliminary analysis of striatal LFPs revealed aberrant behavior-related oscillations in the delta to theta range and in gamma activity. Collectively, our results indicate that disrupted corticostriatal processing occurs across multiple HD models despite differences in the severity of the behavioral phenotype. Efforts aimed at normalizing corticostriatal activity may hold the key to developing new HD therapeutics. PMID- 21629719 TI - Gene expression profiling and molecular characterization of antimony resistance in Leishmania amazonensis. AB - BACKGROUND: Drug resistance is a major problem in leishmaniasis chemotherapy. RNA expression profiling using DNA microarrays is a suitable approach to study simultaneous events leading to a drug-resistance phenotype. Genomic analysis has been performed primarily with Old World Leishmania species and here we investigate molecular alterations in antimony resistance in the New World species L. amazonensis. METHODS/PRINCIPAL FINDINGS: We selected populations of L. amazonensis promastigotes for resistance to antimony by step-wise drug pressure. Gene expression of highly resistant mutants was studied using DNA microarrays. RNA expression profiling of antimony-resistant L. amazonensis revealed the overexpression of genes involved in drug resistance including the ABC transporter MRPA and several genes related to thiol metabolism. The MRPA overexpression was validated by quantitative real-time RT-PCR and further analysis revealed that this increased expression was correlated to gene amplification as part of extrachromosomal linear amplicons in some mutants and as part of supernumerary chromosomes in other mutants. The expression of several other genes encoding hypothetical proteins but also nucleobase and glucose transporter encoding genes were found to be modulated. CONCLUSIONS/SIGNIFICANCE: Mechanisms classically found in Old World antimony resistant Leishmania were also highlighted in New World antimony-resistant L. amazonensis. These studies were useful to the identification of resistance molecular markers. PMID- 21629720 TI - PCR-based identification of Klebsiella pneumoniae subsp. rhinoscleromatis, the agent of rhinoscleroma. AB - Rhinoscleroma is a chronic granulomatous infection of the upper airways caused by the bacterium Klebsiella pneumoniae subsp. rhinoscleromatis. The disease is endemic in tropical and subtropical areas, but its diagnosis remains difficult. As a consequence, and despite available antibiotherapy, some patients evolve advanced stages that can lead to disfiguration, severe respiratory impairment and death by anoxia. Because identification of the etiologic agent is crucial for the definitive diagnosis of the disease, the aim of this study was to develop two simple PCR assays. We took advantage of the fact that all Klebsiella pneumoniae subsp. rhinoscleromatis isolates are (i) of capsular serotype K3; and (ii) belong to a single clone with diagnostic single nucleotide polymorphisms (SNP). The complete sequence of the genomic region comprising the capsular polysaccharide synthesis (cps) gene cluster was determined. Putative functions of the 21 genes identified were consistent with the structure of the K3 antigen. The K3-specific sequence of gene Kr11509 (wzy) was exploited to set up a PCR test, which was positive for 40 K3 strains but negative when assayed on the 76 other Klebsiella capsular types. Further, to discriminate Klebsiella pneumoniae subsp. rhinoscleromatis from other K3 Klebsiella strains, a specific PCR assay was developed based on diagnostic SNPs in the phosphate porin gene phoE. This work provides rapid and simple molecular tools to confirm the diagnostic of rhinoscleroma, which should improve patient care as well as knowledge on the prevalence and epidemiology of rhinoscleroma. PMID- 21629721 TI - Latent microsporidial infection in immunocompetent individuals - a longitudinal study. AB - BACKGROUND: Microsporidia (Fungi) have been repeatedly identified as the cause of opportunistic infections predominantly in immunodeficient individuals such as AIDS patients. However, the global epidemiology of human microsporidiosis is poorly understood and the ability of microsporidia to survive and multiply in immunocompetent hosts remains unsolved. AIMS: To determine the presence of latent microsporidia infections in apparently healthy humans in the Czech Republic, the authors tested sera, urine and stool originating from fifteen persons within a three month period examined on a weekly basis. METHODS: Sera, stool and urine samples originating from fifteen HIV-negative people at risk with occupational exposure to animals, aged 22-56 years, living in the Czech Republic were tested by indirect immunofluorescence assay (IFA) for the presence of specific anti microsporidial antibodies, standard Calcofluor M2R staining for the detection of microsporidian spores in all urine sediments and stool smears and molecular methods for the microsporidial species determination. RESULTS: Specific anti microsporidial antibodies were detected in fourteen individuals, asymptomatic Encephalitozoon spp. infection was found in thirteen and E. bieneusi infection was detected in seven of those examined. While E. hellem 1A and E. cuniculi II were the major causative agents identified, seven different genotypes of E. bieneusi were recorded. CONCLUSIONS: These findings clearly show that exposure to microsporidia is common and chronic microsporidiosis is not linked to any clinical manifestation in healthy population. Moreover, our results indicate much higher incidence of microsporidial infections among an apparently healthy population than previously reported. These results open the question about the potential risk of reactivation of latent microsporidiosis in cases of immunosupression causing life-threatening disease. PMID- 21629722 TI - Clinical manifestations associated with neurocysticercosis: a systematic review. AB - BACKGROUND: The clinical manifestations of neurocysticercosis (NCC) are poorly understood. This systematic review aims to estimate the frequencies of different manifestations, complications and disabilities associated with NCC. METHODS: A systematic search of the literature published from January 1, 1990, to June 1, 2008, in 24 different electronic databases and 8 languages was conducted. Meta analyses were conducted when appropriate. RESULTS: A total of 1569 documents were identified, and 21 included in the analysis. Among patients seen in neurology clinics, seizures/epilepsy were the most common manifestations (78.8%, 95%CI: 65.1%-89.7%) followed by headaches (37.9%, 95%CI: 23.3%-53.7%), focal deficits (16.0%, 95%CI: 9.7%-23.6%) and signs of increased intracranial pressure (11.7%, 95%CI: 6.0%-18.9%). All other manifestations occurred in less than 10% of symptomatic NCC patients. Only four studies reported on the mortality rate of NCC. CONCLUSIONS: NCC is a pleomorphic disease linked to a range of manifestations. Although definitions of manifestations were very rarely provided, and varied from study to study, the proportion of NCC cases with seizures/epilepsy and the proportion of headaches were consistent across studies. These estimates are only applicable to patients who are ill enough to seek care in neurology clinics and likely over estimate the frequency of manifestations among all NCC cases. PMID- 21629723 TI - Crystal structures of T. b. rhodesiense adenosine kinase complexed with inhibitor and activator: implications for catalysis and hyperactivation. AB - BACKGROUND: The essential purine salvage pathway of Trypanosoma brucei bears interesting catalytic enzymes for chemotherapeutic intervention of Human African Trypanosomiasis. Unlike mammalian cells, trypanosomes lack de novo purine synthesis and completely rely on salvage from their hosts. One of the key enzymes is adenosine kinase which catalyzes the phosphorylation of ingested adenosine to form adenosine monophosphate (AMP) utilizing adenosine triphosphate (ATP) as the preferred phosphoryl donor. METHODS AND FINDINGS: Here, we present the first structures of Trypanosoma brucei rhodesiense adenosine kinase (TbrAK): the structure of TbrAK in complex with the bisubstrate inhibitor P(1),P(5) di(adenosine-5')-pentaphosphate (AP5A) at 1.55 A, and TbrAK complexed with the recently discovered activator 4-[5-(4-phenoxyphenyl)-2H-pyrazol-3-yl]morpholine (compound 1) at 2.8 A resolution. CONCLUSIONS: The structural details and their comparison give new insights into substrate and activator binding to TbrAK at the molecular level. Further structure-activity relationship analyses of a series of derivatives of compound 1 support the observed binding mode of the activator and provide a possible mechanism of action with respect to their activating effect towards TbrAK. PMID- 21629724 TI - Characterization of the phytochelatin synthase of Schistosoma mansoni. AB - Treatment for schistosomiasis, which is responsible for more than 280,000 deaths annually, depends exclusively on the use of praziquantel. Millions of people are treated annually with praziquantel and drug resistant parasites are likely to evolve. In order to identify novel drug targets the Schistosoma mansoni sequence databases were queried for proteins involved in glutathione metabolism. One potential target identified was phytochelatin synthase (PCS). Phytochelatins are oligopeptides synthesized enzymatically from glutathione by PCS that sequester toxic heavy metals in many organisms. However, humans do not have a PCS gene and do not synthesize phytochelatins. In this study we have characterized the PCS of S. mansoni (SmPCS). The conserved catalytic triad of cysteine-histidine-aspartate found in PCS proteins and cysteine proteases is also found in SmPCS, as are several cysteine residues thought to be involved in heavy metal binding and enzyme activation. The SmPCS open reading frame is considerably extended at both the N- and C-termini compared to PCS from other organisms. Multiple PCS transcripts are produced from the single encoded gene by alternative splicing, resulting in both mitochondrial and cytoplasmic protein variants. Expression of SmPCS in yeast increased cadmium tolerance from less than 50 uM to more than 1,000 uM. We confirmed the function of SmPCS by identifying PCs in yeast cell extracts using HPLC-mass spectrometry. SmPCS was found to be expressed in all mammalian stages of worm development investigated. Increases in SmPCS expression were seen in ex vivo worms cultured in the presence of iron, copper, cadmium, or zinc. Collectively, these results indicate that SmPCS plays an important role in schistosome response to heavy metals and that PCS is a potential drug target for schistosomiasis treatment. This is the first characterization of a PCS from a parasitic organism. PMID- 21629725 TI - Genetic reconstruction of protozoan rRNA decoding sites provides a rationale for paromomycin activity against Leishmania and Trypanosoma. AB - Aminoglycoside antibiotics target the ribosomal decoding A-site and are active against a broad spectrum of bacteria. These compounds bind to a highly conserved stem-loop-stem structure in helix 44 of bacterial 16S rRNA. One particular aminoglycoside, paromomycin, also shows potent antiprotozoal activity and is used for the treatment of parasitic infections, e.g. by Leishmania spp. The precise drug target is, however, unclear; in particular whether aminoglycoside antibiotics target the cytosolic and/or the mitochondrial protozoan ribosome. To establish an experimental model for the study of protozoan decoding-site function, we constructed bacterial chimeric ribosomes where the central part of bacterial 16S rRNA helix 44 has been replaced by the corresponding Leishmania and Trypanosoma rRNA sequences. Relating the results from in-vitro ribosomal assays to that of in-vivo aminoglycoside activity against Trypanosoma brucei, as assessed in cell cultures and in a mouse model of infection, we conclude that aminoglycosides affect cytosolic translation while the mitochondrial ribosome of trypanosomes is not a target for aminoglycoside antibiotics. PMID- 21629726 TI - Polyparasitism is associated with increased disease severity in Toxoplasma gondii infected marine sentinel species. AB - In 1995, one of the largest outbreaks of human toxoplasmosis occurred in the Pacific Northwest region of North America. Genetic typing identified a novel Toxoplasma gondii strain linked to the outbreak, in which a wide spectrum of human disease was observed. For this globally-distributed, water-borne zoonosis, strain type is one variable influencing disease, but the inability of strain type to consistently explain variations in disease severity suggests that parasite genotype alone does not determine the outcome of infection. We investigated polyparasitism (infection with multiple parasite species) as a modulator of disease severity by examining the association of concomitant infection of T. gondii and the related parasite Sarcocystis neurona with protozoal disease in wild marine mammals from the Pacific Northwest. These hosts ostensibly serve as sentinels for the detection of terrestrial parasites implicated in water-borne epidemics of humans and wildlife in this endemic region. Marine mammals (151 stranded and 10 healthy individuals) sampled over 6 years were assessed for protozoal infection using multi-locus PCR-DNA sequencing directly from host tissues. Genetic analyses uncovered a high prevalence and diversity of protozoa, with 147/161 (91%) of our sampled population infected. From 2004 to 2009, the relative frequency of S. neurona infections increased dramatically, surpassing that of T. gondii. The majority of T. gondii infections were by genotypes bearing Type I lineage alleles, though strain genotype was not associated with disease severity. Significantly, polyparasitism with S. neurona and T. gondii was common (42%) and was associated with higher mortality and more severe protozoal encephalitis. Our finding of widespread polyparasitism among marine mammals indicates pervasive contamination of waterways by zoonotic agents. Furthermore, the significant association of concomitant infection with mortality and protozoal encephalitis identifies polyparasitism as an important factor contributing to disease severity in marine mammals. PMID- 21629727 TI - Host alternation is necessary to maintain the genome stability of rift valley fever virus. AB - BACKGROUND: Most arthropod-borne viruses (arboviruses) are RNA viruses, which are maintained in nature by replication cycles that alternate between arthropod and vertebrate hosts. Arboviruses appear to experience lower rates of evolution than RNA viruses that replicate in a single host. This genetic stability is assumed to result from a fitness trade-off imposed by host alternation, which constrains arbovirus genome evolution. To test this hypothesis, we used Rift Valley fever virus (RVFV), an arbovirus that can be transmitted either directly (between vertebrates during the manipulation of infected tissues, and between mosquitoes by vertical transmission) or indirectly (from one vertebrate to another by mosquito-borne transmission). METHODOLOGY/PRINCIPAL FINDINGS: RVFV was serially passaged in BHK21 (hamster) or Aag2 (Aedes aegypti) cells, or in alternation between the two cell types. After 30 passages, these single host-passaged viruses lost their virulence and induced protective effects against a challenge with a virulent virus. Large deletions in the NSs gene that encodes the virulence factor were detectable from the 15(th) serial passage onwards in BHK21 cells and from the 10(th) passage in Aag2 cells. The phosphoprotein NSs is not essential to viral replication allowing clones carrying deletions in NSs to predominate as they replicate slightly more rapidly. No genetic changes were found in viruses that were passaged alternately between arthropod and vertebrate cells. Furthermore, alternating passaged viruses presenting complete NSs gene remained virulent after 30 passages. CONCLUSIONS/SIGNIFICANCE: Our results strongly support the view that alternating replication is necessary to maintain the virulence factor carried by the NSs phosphoprotein. PMID- 21629729 TI - Comparison of argentinean saint louis encephalitis virus non-epidemic and epidemic strain infections in an avian model. AB - St. Louis encephalitis virus (SLEV, Flavivirus, Flaviviridae) is an emerging mosquito-borne pathogen in South America, with human SLEV encephalitis cases reported in Argentina and Brazil. Genotype III strains of SLEV were isolated from Culex quinquefasciatus mosquitoes in Cordoba, Argentina in 2005, during the largest SLEV outbreak ever reported in South America. The present study tested the hypothesis that the recent, epidemic SLEV strain exhibits greater virulence in birds as compared with a non-epidemic genotype III strain isolated from mosquitoes in Santa Fe Province 27 years earlier. The observed differences in infection parameters between adult House sparrows (Passer domesticus) that were needle-inoculated with either the epidemic or historic SLEV strain were not statistically significant. However, only the House sparrows that were infected with the epidemic strain achieved infectious-level viremia titers sufficient to infect Cx. spp. mosquitoes vectors. Furthermore, the vertebrate reservoir competence index values indicated an approximately 3-fold increase in amplification potential of House sparrows infected with the epidemic strain when pre-existing flavivirus-reactive antibodies were present, suggesting the possibility that antibody-dependent enhancement may increase the risk of avian amplified transmission of SLEV in South America. PMID- 21629728 TI - Tissue and stage-specific distribution of Wolbachia in Brugia malayi. AB - BACKGROUND: Most filarial parasite species contain Wolbachia, obligatory bacterial endosymbionts that are crucial for filarial development and reproduction. They are targets for alternative chemotherapy, but their role in the biology of filarial nematodes is not well understood. Light microscopy provides important information on morphology, localization and potential function of these bacteria. Surprisingly, immunohistology and in situ hybridization techniques have not been widely used to monitor Wolbachia distribution during the filarial life cycle. METHODS/PRINCIPAL FINDINGS: A monoclonal antibody directed against Wolbachia surface protein and in situ hybridization targeting Wolbachia 16S rRNA were used to monitor Wolbachia during the life cycle of B. malayi. In microfilariae and vector stage larvae only a few cells contain Wolbachia. In contrast, large numbers of Wolbachia were detected in the lateral chords of L4 larvae, but no endobacteria were detected in the genital primordium. In young adult worms (5 weeks p.i.), a massive expansion of Wolbachia was observed in the lateral chords adjacent to ovaries or testis, but no endobacteria were detected in the growth zone of the ovaries, uterus, the growth zone of the testis or the vas deferens. Confocal laser scanning and transmission electron microscopy showed that numerous Wolbachia are aligned towards the developing ovaries and single endobacteria were detected in the germline. In inseminated females (8 weeks p.i.) Wolbachia were observed in the ovaries, embryos and in decreasing numbers in the lateral chords. In young males Wolbachia were found in distinct zones of the testis and in large numbers in the lateral chords in the vicinity of testicular tissue but never in mature spermatids or spermatozoa. CONCLUSIONS: Immunohistology and in situ hybridization show distinct tissue and stage specific distribution patterns for Wolbachia in B. malayi. Extensive multiplication of Wolbachia occurs in the lateral chords of L4 and young adults adjacent to germline cells. PMID- 21629730 TI - The use of a mobile laboratory unit in support of patient management and epidemiological surveillance during the 2005 Marburg Outbreak in Angola. AB - BACKGROUND: Marburg virus (MARV), a zoonotic pathogen causing severe hemorrhagic fever in man, has emerged in Angola resulting in the largest outbreak of Marburg hemorrhagic fever (MHF) with the highest case fatality rate to date. METHODOLOGY/PRINCIPAL FINDINGS: A mobile laboratory unit (MLU) was deployed as part of the World Health Organization outbreak response. Utilizing quantitative real-time PCR assays, this laboratory provided specific MARV diagnostics in Uige, the epicentre of the outbreak. The MLU operated over a period of 88 days and tested 620 specimens from 388 individuals. Specimens included mainly oral swabs and EDTA blood. Following establishing on site, the MLU operation allowed a diagnostic response in <4 hours from sample receiving. Most cases were found among females in the child-bearing age and in children less than five years of age. The outbreak had a high number of paediatric cases and breastfeeding may have been a factor in MARV transmission as indicated by the epidemiology and MARV positive breast milk specimens. Oral swabs were a useful alternative specimen source to whole blood/serum allowing testing of patients in circumstances of resistance to invasive procedures but limited diagnostic testing to molecular approaches. There was a high concordance in test results between the MLU and the reference laboratory in Luanda operated by the US Centers for Disease Control and Prevention. CONCLUSIONS/SIGNIFICANCE: The MLU was an important outbreak response asset providing support in patient management and epidemiological surveillance. Field laboratory capacity should be expanded and made an essential part of any future outbreak investigation. PMID- 21629731 TI - Economic impact of cystic echinococcosis in peru. AB - BACKGROUND: Cystic echinococcosis (CE) constitutes an important public health problem in Peru. However, no studies have attempted to estimate the monetary and non-monetary impact of CE in Peruvian society. METHODS: We used official and published sources of epidemiological and economic information to estimate direct and indirect costs associated with livestock production losses and human disease in addition to surgical CE-associated disability adjusted life years (DALYs) lost. FINDINGS: The total estimated cost of human CE in Peru was U.S.$2,420,348 (95% CI:1,118,384-4,812,722) per year. Total estimated livestock-associated costs due to CE ranged from U.S.$196,681 (95% CI:141,641-251,629) if only direct losses (i.e., cattle and sheep liver destruction) were taken into consideration to U.S.$3,846,754 (95% CI:2,676,181-4,911,383) if additional production losses (liver condemnation, decreased carcass weight, wool losses, decreased milk production) were accounted for. An estimated 1,139 (95% CI: 861-1,489) DALYs were also lost due to surgical cases of CE. CONCLUSIONS: This preliminary and conservative assessment of the socio-economic impact of CE on Peru, which is based largely on official sources of information, very likely underestimates the true extent of the problem. Nevertheless, these estimates illustrate the negative economic impact of CE in Peru. PMID- 21629733 TI - Playing TAG with a bacterial stress response. PMID- 21629732 TI - Metabolic regulation of mycobacterial growth and antibiotic sensitivity. AB - Treatment of chronic bacterial infections, such as tuberculosis (TB), requires a remarkably long course of therapy, despite the availability of drugs that are rapidly bacteriocidal in vitro. This observation has long been attributed to the presence of bacterial populations in the host that are "drug-tolerant" because of their slow replication and low rate of metabolism. However, both the physiologic state of these hypothetical drug-tolerant populations and the bacterial pathways that regulate growth and metabolism in vivo remain obscure. Here we demonstrate that diverse growth-limiting stresses trigger a common signal transduction pathway in Mycobacterium tuberculosis that leads to the induction of triglyceride synthesis. This pathway plays a causal role in reducing growth and antibiotic efficacy by redirecting cellular carbon fluxes away from the tricarboxylic acid cycle. Mutants in which this metabolic switch is disrupted are unable to arrest their growth in response to stress and remain sensitive to antibiotics during infection. Thus, this regulatory pathway contributes to antibiotic tolerance in vivo, and its modulation may represent a novel strategy for accelerating TB treatment. PMID- 21629735 TI - The burden of selected chronic non-communicable diseases and their risk factors in Malawi: nationwide STEPS survey. AB - BACKGROUND: Chronic non-communicable diseases (NCDs) are becoming significant causes of morbidity and mortality, particularly in sub-Saharan African countries, although local, high-quality data to inform evidence-based policies are lacking. OBJECTIVES: To determine the magnitude of NCDs and their risk factors in Malawi. METHODS: Using the WHO STEPwise approach to chronic disease risk factor surveillance, a population-based, nationwide cross-sectional survey was conducted between July and September 2009 on participants aged 25-64 years. Socio demographic and behaviour risk factors were collected in Step 1. Physical anthropometric measurements and blood pressure were documented in Step 2. Blood cholesterol and fasting blood glucose were measured in Step 3. RESULTS AND CONCLUSION: A total of 5,206 adults (67% females) were surveyed. Tobacco smoking, alcohol drinking and raised blood pressure (BP) were more frequent in males than females, 25% vs 3%, 30% vs 4% and 37% vs 29%. Overweight, physical inactivity and raised cholesterol were more common in females than males, 28% vs 16%, 13% vs 6% and 11% vs 6%. Tobacco smoking was more common in rural than urban areas 11% vs 7%, and overweight and physical inactivity more common in urban than rural areas 39% vs 22% and 24% vs 9%, all with p<0.05. Overall (both sexes) prevalence of tobacco smoking, alcohol consumption, overweight and physical inactivity was 14%, 17%, 22%, 10% and prevalence of raised BP, fasting blood sugar and cholesterol was 33%, 6% and 9% respectively. These data could be useful in the formulation and advocacy of NCD policy and action plan in Malawi. PMID- 21629734 TI - Targeting the DNA double strand break repair machinery in prostate cancer. AB - Regardless of the achievable remissions with first line hormone therapy in patients with prostate cancer (CaP), the disease escapes the hormone dependent stage to a more aggressive status where chemotherapy is the only effective treatment and no treatment is curative. This makes it very important to identify new targets that can improve the outcome of treatment. ATM and DNA-PK are the two kinases responsible for signalling and repairing double strand breaks (DSB). Thus, both kinases are pertinent targets in CaP treatment to enhance the activity of the numerous DNA DSB inducing agents used in CaP treatment such as ionizing radiation (IR). Colony formation assay was used to assess the sensitivity of hormone dependent, p53 wt (LNCaP) and hormone independent p53 mutant (PC3) CaP cell lines to the cytotoxic effect of IR and Doxorubicin in the presence or absence of Ku55933 and NU7441 which are small molecule inhibitors of ATM and DNA PK, respectively. Flow cytometry based methods were used to assess the effect of the two inhibitors on cell cycle, apoptosis and H2AX foci formation. Neutral comet assay was used to assess the induction of DNA DSBs. Ku55933 or NU7441 alone increased the sensitivity of CaP cell lines to the DNA damaging agents, however combining both inhibitors together resulted in further enhancement of sensitivity. The cell cycle profile of both cell lines was altered with increased cell death, DNA DSBs and H2AX foci formation. This study justifies further evaluation of the ATM and DNA-PK inhibitors for clinical application in CaP patients. Additionally, the augmented effect resulting from combining both inhibitors may have a significant implication for the treatment of CaP patients who have a defect in one of the two DSB repair pathways. PMID- 21629736 TI - Glycogen synthase kinase-3beta: a mediator of inflammation in Alzheimer's disease? AB - Proliferation and activation of microglial cells is a neuropathological characteristic of brain injury and neurodegeneration, including Alzheimer's disease. Microglia act as the first and main form of immune defense in the nervous system. While the primary function of microglia is to survey and maintain the cellular environment optimal for neurons in the brain parenchyma by actively scavenging the brain for damaged brain cells and foreign proteins or particles, sustained activation of microglia may result in high production of proinflammatory mediators that disturb normal brain functions and even cause neuronal injury. Glycogen synthase kinase-3beta has been recently identified as a major regulator of immune system and mediates inflammatory responses in microglia. Glycogen synthase kinase-3beta has been extensively investigated in connection to tau and amyloid beta toxicity, whereas reports on the role of this enzyme in neuroinflammation in Alzheimer's disease are negligible. Here we review and discuss the role of glycogen synthase-3beta in immune cells in the context of Alzheimer's disease pathology. PMID- 21629737 TI - Self-Renewal Signalling in Presenescent Tetraploid IMR90 Cells. AB - Endopolyploidy and genomic instability are shared features of both stress-induced cellular senescence and malignant growth. Here, we examined these facets in the widely used normal human fibroblast model of senescence, IMR90. At the presenescence stage, a small (2-7%) proportion of cells overcome the 4n-G1 checkpoint, simultaneously inducing self-renewal (NANOG-positivity), the DNA damage response (DDR; gamma-H2AX-positive foci), and senescence (p16inka4a- and p21CIP1-positivity) signalling, some cells reach octoploid DNA content and divide. All of these markers initially appear and partially colocalise in the perinucleolar compartment. Further, with development of senescence and accumulation of p16inka4a and p21CIP1, NANOG is downregulated in most cells. The cells increasingly arrest in the 4n-G1 fraction, completely halt divisions and ultimately degenerate. A positive link between DDR, self-renewal, and senescence signalling is initiated in the cells overcoming the tetraploidy barrier, indicating that cellular and molecular context of induced tetraploidy during this period of presenescence is favourable for carcinogenesis. PMID- 21629738 TI - GSK-3 in Neurodegenerative Diseases. AB - Glycogen synthase kinase-3 (GSK-3) regulates multiple cellular processes, and its dysregulation is implicated in the pathogenesis of diverse diseases. In this paper we will focus on the dysfunction of GSK-3 in Alzheimer's disease and Parkinson's disease. Specifically, GSK-3 is known to interact with tau, beta amyloid (Abeta), and alpha-synuclein, and as such may be crucially involved in both diseases. Abeta production, for example, is regulated by GSK-3, and its toxicity is mediated by GSK-induced tau phosphorylation and degeneration. alpha synuclein is a substrate for GSK-3 and GSK-3 inhibition protects against Parkinsonian toxins. Lithium, a GSK-3 inhibitor, has also been shown to affect tau, Abeta, and alpha-synuclein in cell culture, and transgenic animal models. Thus, understanding the role of GSK-3 in neurodegenerative diseases will enhance our understanding of the basic mechanisms underlying the pathogenesis of these disorders and also facilitate the identification of new therapeutic avenues. PMID- 21629740 TI - End of life: a family narrative. AB - This paper is based on ethnographic research that examines family reaction to an elderly husband and father's end of life. From a group of 30 families in our study (family defined as a widow aged 70 and over and two adult biological children between the ages of 40 and 60), we offer an extreme case example of family bereavement. We report our findings through the open-ended responses of a widow and two children who were interviewed ten months after the death of the husband and father. Three general themes emerged: (1) how the family imputes meaning to the end of life, (2) changes in the roles of family members, and (3) the family's ways of coping with the death, particularly through their belief system. A key finding is that the meaning family members find in their loved one's death is tied to the context of his death (how and where he died), their perception of his quality of life as a whole, and their philosophical, religious, and spiritual beliefs about life, death, and the afterlife that are already in place. PMID- 21629739 TI - Acute myeloid leukemia with the t(8;21) translocation: clinical consequences and biological implications. AB - The t(8;21) abnormality occurs in a minority of acute myeloid leukemia (AML) patients. The translocation results in an in-frame fusion of two genes, resulting in a fusion protein of one N-terminal domain from the AML1 gene and four C terminal domains from the ETO gene. This protein has multiple effects on the regulation of the proliferation, the differentiation, and the viability of leukemic cells. The translocation can be detected as the only genetic abnormality or as part of more complex abnormalities. If t(8;21) is detected in a patient with bone marrow pathology, the diagnosis AML can be made based on this abnormality alone. t(8;21) is usually associated with a good prognosis. Whether the detection of the fusion gene can be used for evaluation of minimal residual disease and risk of leukemia relapse remains to be clarified. To conclude, detection of t(8;21) is essential for optimal handling of these patients as it has both diagnostic, prognostic, and therapeutic implications. PMID- 21629742 TI - Rapid assessment of age-related differences in standing balance. AB - As life expectancy continues to rise, in the future there will be an increasing number of older people prone to falling. Accordingly, there is an urgent need for comprehensive testing of older individuals to collect data and to identify possible risk factors for falling. Here we use a low-cost force platform to rapidly assess deficits in balance under various conditions. We tested 21 healthy older adults and 24 young adults during static stance, unidirectional and rotational displacement of their centre of pressure (COP). We found an age related increase in postural sway during quiet standing and a reduction of maximal COP displacement in unidirectional and rotational displacement tests. Our data show that even low-cost computerized assessment tools allow for the comprehensive testing of balance performance in older subjects. PMID- 21629743 TI - Centella asiatica Attenuates D-Galactose-Induced Cognitive Impairment, Oxidative and Mitochondrial Dysfunction in Mice. AB - D-galactose induced neurotoxicity is well known model for studying aging and related oxidative damage and memory impairment. Aging is a biological process, characterized by the gradual loss of physiological functions by unknown mechanism. Centella asiatica, Indian pennywort has been documented in the treatment of various neurological disorders including aging. Therefore, present study has been conducted in order to explore the possible role of Centella asiatica against D-galactose induced cognitive impairment, oxidative and mitochondrial dysfunction in mice. Chronic administration of D-galactose (100 mg/kg s.c.) for a period of six weeks significantly impaired cognitive task (both in both Morris water maze and elevated plus maze) and oxidative defense (Increased lipid peroxidation, nitrite concentration and decreased activity of superoxide dismutase, catalase and non-protein thiols) and impaired mitochondrial complex (I, II and III) enzymes activities as compared to sham group. Six weeks Centella asiatica (150 and 300 mg/kg, p.o) treatment significantly improved behavioral alterations, oxidative damage and mitochondrial enzyme complex activities as compared to contro l (D-galactose). Centella asiatica also attenuated enhanced acetylcholine esterase enzyme level in D-galactose senescence mice. Present study highlights the protective effect of Centella asiatica against D-galactose induced behavioral, biochemical and mitochondrial dysfunction in mice. PMID- 21629741 TI - Melatonin in mitochondrial dysfunction and related disorders. AB - Mitochondrial dysfunction is considered one of the major causative factors in the aging process, ischemia/reperfusion (I/R), septic shock, and neurodegenerative disorders like Parkinson's disease (PD), Alzheimer's disease (AD), and Huntington's disease (HD). Increased free radical generation, enhanced mitochondrial inducible nitric oxide (NO) synthase activity, enhanced NO production, decreased respiratory complex activity, impaired electron transport system, and opening of mitochondrial permeability transition pore all have been suggested as factors responsible for impaired mitochondrial function. Melatonin, the major hormone of the pineal gland, also acts as an antioxidant and as a regulator of mitochondrial bioenergetic function. Both in vitro and in vivo, melatonin was effective for preventing oxidative stress/nitrosative stress induced mitochondrial dysfunction seen in experimental models of PD, AD, and HD. In addition, melatonin is known to retard aging and to inhibit the lethal effects of septic shock or I/R lesions by maintaining respiratory complex activities, electron transport chain, and ATP production in mitochondria. Melatonin is selectively taken up by mitochondrial membranes, a function not shared by other antioxidants. Melatonin has thus emerged as a major potential therapeutic tool for treating neurodegenerative disorders such as PD or AD, and for preventing the lethal effects of septic shock or I/R. PMID- 21629744 TI - Myelodysplastic syndrome and histone deacetylase inhibitors: "to be or not to be acetylated"? AB - Myelodysplastic syndrome (MDS) represents a heterogeneous group of diseases with clonal proliferation, bone marrow failure and increasing risk of transformation into an acute myeloid leukaemia. Structured guidelines are developed for selective therapy based on prognostic subgroups, age, and performance status. Although many driving forces of disease phenotype and biology are described, the complete and possibly interacting pathogenetic pathways still remain unclear. Epigenetic investigations of cancer and haematologic diseases like MDS give new insights into the pathogenesis of this complex disease. Modifications of DNA or histones via methylation or acetylation lead to gene silencing and altered physiology relevant for MDS. First clinical trials give evidence that patients with MDS could benefit from epigenetic treatment with, for example, DNA methyl transferase inhibitors (DNMTi) or histone deacetylase inhibitors (HDACi). Nevertheless, many issues of HDACi remain incompletely understood and pose clinical and translational challenges. In this paper, major aspects of MDS, MDS associated epigenetics and the potential use of HDACi are discussed. PMID- 21629745 TI - The structural basis of ligand recognition by natural killer cell receptors. AB - Natural killer cells are a group of lymphocytes which function as tightly controlled surveillance operatives which identify transformed cells through a discrete balance of activating and inhibitory receptors ultimately leading to the destruction of incongruent cells. The understanding of this finely tuned balancing act has been aided by the high-resolution structure determination of activating and inhibitory receptors both alone and in complex with their ligands. This paper collates these structural studies detailing the aspects which directly relate to the natural killer cell function and serves to inform both the specialized structural biologist reader and a more general immunology audience. PMID- 21629746 TI - MicroRNAs Regulate Key Effector Pathways of Senescence. AB - MicroRNAs (miRNAs) are small (approximately 22 nt) noncoding endogenous RNA molecules that regulate gene expression and protein coding by base pairing with the 3' untranslated region (UTR) of target mRNAs. miRNA expression is associated with cancer pathogenesis because miRNAs are intimately linked to cancer development. Senescence blocks cell proliferation, representing an important barrier that cells must bypass to reach malignancy. Importantly, certain miRNAs have been shown to have an important role during cellular senescence, which is also involved in human tumorigenesis. Therefore, therapeutic induction of senescence by drugs or miRNA-based therapies is a potential method to treat cancer by inducing a persistent growth arrest in tumors. PMID- 21629747 TI - Deconstructing GSK-3: The Fine Regulation of Its Activity. AB - Glycogen synthase kinase-3 (GSK-3) unique position in modulating the function of a diverse series of proteins in combination with its association with a wide variety of human disorders has attracted significant attention to the protein both as a therapeutic target and as a means to understand the molecular basis of these disorders. GSK-3 is ubiquitously expressed and, unusually, constitutively active in resting, unstimulated cells. In mammals, GSK-3alpha and beta are each expressed widely at both the RNA and protein levels although some tissues show preferential levels of some of the two proteins. Neither gene appears to be acutely regulated at the transcriptional level, whereas the proteins are controlled posttranslationally, largely through protein-protein interactions or by posttranslational regulation. Control of GSK-3 activity thus occurs by complex mechanisms that are each dependent upon specific signalling pathways. Furthermore, GSK-3 appears to be a cellular nexus, integrating several signalling systems, including several second messengers and a wide selection of cellular stimulants. This paper will focus on the different ways to control GSK-3 activity (phosphorylation, protein complex formation, truncation, subcellular localization, etc.), the main signalling pathways involved in its control, and its pathological deregulation. PMID- 21629748 TI - Eicosanoid Production following One Bout of Exercise in Middle-Aged African American Pre- and Stage 1 Hypertensives. AB - Endothelial dysfunction and a sedentary lifestyle may be involved in the development of hypertension which is proliferative among middle-aged African Americans (AA). Signaling molecules derived from the oxidation of 20-carbon fatty acid molecules known as eicosanoids influence vascular tone. The relationship between aerobic fitness and eicosanoid formation following exercise in middle aged African American hypertensives is unknown. Purpose. To determine the relationship between aerobic capacity and eicosanoid formation after a bout of moderate-intensity exercise in middle-aged AA hypertensives. Methods. Ten sedentary hypertensive AA underwent 50 min of aerobic exercise at 65% VO(2)max. Urine was collected for 24 hr on two occasions, prior to testing and immediately following the bout of exercise. Urinary metabolites of prostacyclin (6-keto PGF(1alpha)) and thromboxane (11-dTXB(2)) were measured during the day and night periods by high-performance liquid chromatography (HPLC). Results. 6-keto PGF(1alpha) levels significantly increased (P = .04) following the bout of exercise compared to the control day. There was a significant relationship (r = .49, P < .05) between 6-keto PGF(1alpha) levels and VO(2)max during the exercise day. Conclusion. Based on this preliminary study, there appears to be a relationship between aerobic capacity and exercise-induced 6-keto PGF(1alpha) production in middle-aged hypertensive AAs. AAs with lower VO(2)max had lower 6 keto PGF(1alpha) formation. PMID- 21629749 TI - Brain functional network in Alzheimer's disease: diagnostic markers for diagnosis and monitoring. AB - Alzheimer's disease (AD) is the most common type of dementia that is clinically characterized by the presence of memory impairment and later by impairment in other cognitive domains. The clinical diagnosis is based on interviews with the patient and his/her relatives and on neuropsychological assessment, which are also used to monitor cognitive decline over time. Several biomarkers have been proposed for detecting AD in its earliest stages, that is, in the predementia stage. In an attempt to find noninvasive biomarkers, researchers have investigated the feasibility of neuroimaging tools, such as MR, SPECT, and FDG PET imaging, as well as neurophysiological measurements using EEG. In this paper, we investigate the brain functional networks in AD, focusing on main neurophysiological techniques, integrating with most relevant functional brain imaging findings. PMID- 21629751 TI - A simple and fast method for the production and characterization of methylic and ethylic biodiesels from tucum oil via an alkaline route. AB - A simple, fast, and complete route for the production of methylic and ethylic biodiesel from tucum oil is described. Aliquots of the oil obtained directly from pressed tucum (pulp and almonds) were treated with potassium methoxide or ethoxide at 40 degrees C for 40 min. The biodiesel form was removed from the reactor and washed with 0.1 M HCl aqueous solution. A simple distillation at 100 degrees C was carried out in order to remove water and alcohol species from the biodiesel. The oxidative stability index was obtained for the tucum oil as well as the methylic and ethylic biodiesel at 6.13, 2.90, and 2.80 h, for storage times higher than 8 days. Quality control of the original oil and of the methylic and ethylic biodiesels, such as the amount of glycerin produced during the transesterification process, was accomplished by the TLC, GC-MS, and FT-IR techniques. The results obtained in this study indicate a potential biofuel production by simple treatment of tucum, an important Amazonian fruit. PMID- 21629750 TI - KIR/HLA interactions and pathogen immunity. AB - The innate immune system is the first line of defence in response to pathogen infection. Natural killer (NK) cells perform a vital role in this response with the ability to directly kill infected cells, produce cytokines, and cross-talk with the adaptive immune system. These effector functions are dependent on activation of NK cells which is determined by surface receptor interactions with ligands on target cells. Of these receptors, the polymorphic killer immunoglobulin-like receptors (KIRs), which interact with MHC class 1 (also highly polymorphic), are largely inhibitory, and exhibit substantial genetic diversity. The result is a significant variation of NK cell repertoire between individuals and also between populations, with a multitude of possible KIR:HLA combinations. As each KIR:ligand interaction may have differential effects on NK cell activation and inhibition, this diversity has important potential influences on the host response to infections. Genetic studies have demonstrated associations between specific KIR:ligand combinations and the outcome of viral (and other) infections, in particular hepatitis C and HIV infection. Detailed functional studies are not required to define the mechanisms underpinning these disease associations. PMID- 21629752 TI - The Ambulatory and Home Care Record: A Methodological Framework for Economic Analyses in End-of-Life Care. AB - Provision of end-of-life care in North America takes place across a multitude of settings, including hospitals, ambulatory clinics and home settings. As a result, family caregiving is characteristically a major component of care within the home. Accordingly, economic evaluation of the end-of-life care environment must devote equal consideration to resources provided by the public health system as well as privately financed resources, such as time and money provided by family caregivers. This paper addresses the methods used to measure end-of-life care costs. The existing empirical literature will be reviewed in order to assess care costs with areas neglected in this body of literature to be identified. The Ambulatory and Home Care Record, a framework and tool for comprehensively measuring costs related to the provision and receipt of end-of-life care across all health care settings, will be described and proposed. Finally, areas for future work will be identified, along with their potential contribution to this body of knowledge. PMID- 21629753 TI - WF10 stimulates NK cell cytotoxicity by increasing LFA-1-mediated adhesion to tumor cells. AB - The redox-active chlorite-based drug WF10 (Immunokine) was shown to have modulatory effects on both the innate and adaptive immune system in vitro and in vivo. Animal studies suggest that WF10 enhances immunity against tumors. One possible explanation for such an effect is that WF10 stimulates natural killer cell cytotoxicity against malignant cells. Here, we show that WF10 regulates human NK cell cytotoxicity in a time-dependent manner, following an S-shaped kinetic with an initial stimulation of activity followed by a decrease in activity relative to the untreated controls. WF10 does not activate NK cells on its own but co-stimulates NK cell activation mediated by different activating receptors. This is mediated by enhancing NK cell adhesion to target cells through promoting the activation of the integrin LFA-1. These data demonstrate a direct effect of WF10 on the cytotoxicity of human NK cells. PMID- 21629755 TI - Faith and end of life in nursing homes. AB - This paper explores the role of religious belief in the experiences of dying and death in a Catholic nursing home. The home appeals to residents and their families due to the active religious presence. Thus, religion is a salient element of the "local culture" which exists in this long-term care setting. The preeminence of faith within the organization and the personal religious convictions of staff, residents, and families may drive how death and dying are discussed and experienced in this setting, as well as the meanings that are attached to them. This paper examines the relationship between faith and the experience and meaning of death in this nursing home. We present themes that emerged from open-ended interviews with residents, family members, and staff, gathered between 1996 and 2004. The data indicate that people select the home due to their Catholic faith and the home's religious tone. Themes also show that belief in God and an afterlife helps shape the experience of dying and death for our informants. Our paper does not compare ease of dying with other nursing homes or within other belief systems. PMID- 21629754 TI - What Are the bona fide GSK3 Substrates? AB - Nearly 100 proteins are proposed to be substrates for GSK3, suggesting that this enzyme is a fundamental regulator of almost every process in the cell, in every tissue in the body. However, it is not certain how many of these proposed substrates are regulated by GSK3 in vivo. Clearly, the identification of the physiological functions of GSK3 will be greatly aided by the identification of its bona fide substrates, and the development of GSK3 as a therapeutic target will be highly influenced by this range of actions, hence the need to accurately establish true GSK3 substrates in cells. In this paper the evidence that proposed GSK3 substrates are likely to be physiological targets is assessed, highlighting the key cellular processes that could be modulated by GSK3 activity and inhibition. PMID- 21629756 TI - Ever-young sex chromosomes in European tree frogs. AB - Non-recombining sex chromosomes are expected to undergo evolutionary decay, ending up genetically degenerated, as has happened in birds and mammals. Why are then sex chromosomes so often homomorphic in cold-blooded vertebrates? One possible explanation is a high rate of turnover events, replacing master sex determining genes by new ones on other chromosomes. An alternative is that X-Y similarity is maintained by occasional recombination events, occurring in sex reversed XY females. Based on mitochondrial and nuclear gene sequences, we estimated the divergence times between European tree frogs (Hyla arborea, H. intermedia, and H. molleri) to the upper Miocene, about 5.4-7.1 million years ago. Sibship analyses of microsatellite polymorphisms revealed that all three species have the same pair of sex chromosomes, with complete absence of X-Y recombination in males. Despite this, sequences of sex-linked loci show no divergence between the X and Y chromosomes. In the phylogeny, the X and Y alleles cluster according to species, not in groups of gametologs. We conclude that sex chromosome homomorphy in these tree frogs does not result from a recent turnover but is maintained over evolutionary timescales by occasional X-Y recombination. Seemingly young sex chromosomes may thus carry old-established sex-determining genes, a result at odds with the view that sex chromosomes necessarily decay until they are replaced. This raises intriguing perspectives regarding the evolutionary dynamics of sexually antagonistic genes and the mechanisms that control X-Y recombination. PMID- 21629757 TI - Employment of oligodeoxynucleotide plus interleukin-2 improves cytogenetic analysis in splenic marginal zone lymphoma. AB - To compare the efficiency of novel mitogenic agents and traditional mitosis inductors, 18 patients with splenic marginal zone lymphoma (SMZL) were studied. Three cultures using oligodeoxynucleotide (ODN) plus interleukin-2 (IL-2), or TPA, or LPS were setup in each patient. Seventeen/18 cases with ODN + IL2 had moderate/good proliferation (94, 4%) as compared with 10/18 cases with TPA and LPS (55%) (P = .015); 14/18 (77, 7%) cases with ODN + IL2 had sufficient good quality of banding as compared with 8/18 cases (44, 4%) with TPA and LPS. The karyotype could be defined from ODN + IL2-stimulated cultures in all 18 patients, 14 of whom (77, 7%) had a cytogenetic aberration, whereas clonal aberrations could be documented in 9 and in 3 cases by stimulation with LPS and TPA, respectively. Recurrent chromosome aberrations in our series were represented by aberrations of chromosome 14q in 5 patients, by trisomy 12 and 7q deletion in 4 cases each, and by abnormalities involving 11q and 13q in two cases each. These findings show that stimulation with ODN + IL2 offers more mitotic figures of better quality and results in an increased rate of clonal aberrations in SMZL, making this method ideal for prospective studies aiming at the definition of the prognostic impact of cytogenetic aberrations in this disorder. PMID- 21629758 TI - Estimating Premorbid Intelligence among Older Adults: The Utility of the AMNART. AB - This study examines the utility of the American version of the National Adult Reading Test (AMNART) as a measure of premorbid intelligence for older adults. In a sample of 130 older adults, aged 56 to 104, the AMNART was compared to other tests of premorbid intelligence. The results revealed that AMNART-estimated IQ was significantly higher than other premorbid estimates. Across specific educational groups (i.e., 0-12, 13-16, and 17 or more years of education), AMNART estimated IQ was inflated relative to all other premorbid estimates. The AMNART also declined as cognitive impairment increased, and there was a significant interaction between aging-related diagnostic group and premorbid estimate. The AMNART may therefore overestimate premorbid ability relative to other premorbid measures, particularly among those with greater cognitive impairment and lower levels of education. These results suggest that the AMNART should be used cautiously among older adults and in conjunction with other estimates of premorbid ability. PMID- 21629759 TI - The use of auditory event-related potentials in Alzheimer's disease diagnosis. AB - Event-related potentials (ERPs) are important clinical and research instruments in neuropsychiatry, particularly due to their strategic role for the investigation of brain function. These techniques are often underutilized in the evaluation of neurological and psychiatric disorders, but ERPs are noninvasive instruments that directly reflect cortical neuronal activity. Previous studies using the P300, P3a, and MMN components of the ERP to study dementing illness are reviewed. The results suggest that particularly the P300 brain potential is sensitive to Alzheimer's disease processes during its early stages, and that easily performed stimulus discrimination tasks are the clinically most useful. Finally, these data suggest that the P300 ERP can aid in the diagnosis of dementia and may help in the assessment of early Alzheimer's disease. PMID- 21629760 TI - The role of proteasome inhibition in nonsmall cell lung cancer. AB - Lung cancer therapy with current available chemotherapeutic agents is mainly palliative. For these and other reasons there is now a great interest to find targeted therapies that can be effective not only palliating lung cancer or decreasing treatment-related toxicity, but also giving hope to cure these patients. It is already well known that the ubiquitin-proteasome system like other cellular pathways is critical for the proliferation and survival of cancer cells; thus, proteosome inhibition has become a very attractive anticancer therapy. There are several phase I and phase II clinical trials now in non-small cell lung cancer and small cell lung cancer using this potential target. Most of the trials use bortezomib in combination with chemotherapeutic agents. This paper tends to make a state-of-the-art review based on the available literature regarding the use of bortezomib as a single agent or in combination with chemotherapy in patients with lung cancer. PMID- 21629761 TI - Recent developments in the remote radio control of insect flight. AB - The continuing miniaturization of digital circuits and the development of low power radio systems coupled with continuing studies into the neurophysiology and dynamics of insect flight are enabling a new class of implantable interfaces capable of controlling insects in free flight for extended periods. We provide context for these developments, review the state-of-the-art and discuss future directions in this field. PMID- 21629762 TI - Aging and the social cognitive determinants of physical activity behavior and behavior change: evidence from the guide to health trial. AB - Part one of this study investigated the effect of aging on social-cognitive characteristics related to physical activity (PA) among adults in the baseline phase of a health promotion intervention. Participants' questionnaire responses and activity logs indicated PA levels and self-efficacy declined with age, while social support and the use of self-regulatory behaviors (e.g., goal setting, planning, and keeping track) increased. With age participants were also less likely to expect PA to interfere with their daily routines and social obligations. Part two of the study was among overweight/obese, inactive participants completing the intervention; it examined whether improvements in psychosocial variables might counteract declining PA associated with age. After treatment, participants were more active and decreased body weight regardless of age, and improved self-efficacy, outcome expectations, and self-regulatory behaviors. In a causal model, increases in self-efficacy at 7-months lead to increased PA levels and, albeit marginally, weight loss at 16 months; increased PA was associated with greater weight loss. Aging adults who were more confident exercised more and as a result lost more weight. This longitudinal study suggests interventions that offset the effect of aging on self-efficacy may be more successful in helping older participants become more active and avoid weight gain. PMID- 21629763 TI - Combining transcranial magnetic stimulation and electroencephalography may contribute to assess the severity of Alzheimer's disease. AB - Alzheimer's disease (AD) is the most common form of old age dementia, and mild cognitive impairment (MCI) often precedes AD. In our previous study (Julkunen et al. 2008), we found that the combination of transcranial magnetic stimulation (TMS) and electroencephalography (EEG) was able to find distinct differences in AD and MCI patients as compared to controls. Here, we reanalyzed the small sample data from our previous study with the aim to test the sensitivity of the TMS-EEG characteristics to discriminate control subjects (n = 4) from MCI (n = 5) and AD (n = 5) subjects. Furthermore, we investigated how the TMS-EEG response characteristics related to the scores of the dementia rating scales used to evaluate the severity of cognitive decline in these subjects. We found that the TMS-EEG response P30 amplitude correlated with cognitive decline and showed good specificity and sensitivity in identifying healthy subjects from those with MCI or AD. Given the small sample size, further studies may be needed to confirm the results. PMID- 21629764 TI - Correlations decrease with propagation of spiking activity in the mouse barrel cortex. AB - Propagation of suprathreshold spiking activity through neuronal populations is important for the function of the central nervous system. Neural correlations have an impact on cortical function particularly on the signaling of information and propagation of spiking activity. Therefore we measured the change in correlations as suprathreshold spiking activity propagated between recurrent neuronal networks of the mammalian cerebral cortex. Using optical methods we recorded spiking activity from large samples of neurons from two neural populations simultaneously. The results indicate that correlations decreased as spiking activity propagated from layer 4 to layer 2/3 in the rodent barrel cortex. PMID- 21629765 TI - Activating transcription factor 3, a useful marker for regenerative response after nerve root injury. AB - Activating transcription factor 3 (ATF3) is induced in various tissues in response to stress. In this experiment, ATF3 expression was studied in adult rats subjected either to a dorsal or ventral root avulsion (VRA; L4-6), or sciatic nerve transection (SNT). Post-operative survival times varied between 1.5 h and 3 weeks. In additional experiments an avulsed ventral root was directly replanted to the spinal cord. Dorsal root ganglias (DRGs) from humans exposed to traumatic dorsal root avulsions were also examined. After SNT ATF3 immunoreactivity (ATF3 IR) was detected in a few DRG neurons already 6 h after the lesion. After 24 h the number had clearly increased and still at 3 weeks DRG neurons remained labeled. In the ventral horn, ATF3 IR in motoneurons (MN) was first detected 24 h after the SNT, and still 3 weeks post-operatively lesioned MN showed ATF3 labeling. After a VRA many spinal MN showed ATF3 IR already after 3 h, and after 6 h all MN were labeled. At 3 weeks a majority of the lesioned MN had died, but all the remaining ones were labeled. When an avulsed ventral root was directly replanted, MN survived and were still labeled at 5 weeks. In DRG, a few neurons were labeled already at 1.5 h after a dorsal root avulsion. At 24 h the number had increased but still only a minority of the neurons were labeled. At 3 days the number of labeled neurons was reduced, and a further reduction was at hand at 7 days and 3 weeks. In parallel, in humans, 3 days after a traumatic dorsal root avulsion, only a few DRG neurons showed ATF3 IR. At 6 weeks no labeled neurons could be detected. These facts imply that ATF3 response to axotomy involves a distance-dependent mechanism. ATF3 also appears to be a useful and reliable neuronal marker of nerve lesions even in humans. In addition, ATF3 up-regulation in both motor and sensory neurons seems to be linked to regenerative competence. PMID- 21629766 TI - Visual mismatch negativity reveals automatic detection of sequential regularity violation. AB - Sequential regularities are abstract rules based on repeating sequences of environmental events, which are useful to make predictions about future events. Here, we tested whether the visual system is capable to detect sequential regularity in unattended stimulus sequences. The visual mismatch negativity (vMMN) component of the event-related potentials is sensitive to the violation of complex regularities (e.g., object-related characteristics, temporal patterns). We used the vMMN component as an index of violation of conditional (if, then) regularities. In the first experiment, to investigate emergence of vMMN and other change-related activity to the violation of conditional rules, red and green disk patterns were delivered in pairs. The majority of pairs comprised of disk patterns with identical colors, whereas in deviant pairs the colors were different. The probabilities of the two colors were equal. The second member of the deviant pairs elicited a vMMN with longer latency and more extended spatial distribution to deviants with lower probability (10 vs. 30%). In the second (control) experiment the emergence of vMMN to violation of a simple, feature related rule was studied using oddball sequences of stimulus pairs where deviant colors were presented with 20% probabilities. Deviant colored patterns elicited a vMMN, and this component was larger for the second member of the pair, i.e., after a shorter inter-stimulus interval. This result corresponds to the SOA/(v)MMN relationship, expected on the basis of a memory-mismatch process. Our results show that the system underlying vMMN is sensitive to abstract, conditional rules. Representation of such rules implicates expectation of a subsequent event, therefore vMMN can be considered as a correlate of violated predictions about the characteristics of environmental events. PMID- 21629767 TI - Blocking Effects of Human Tau on Squid Giant Synapse Transmission and Its Prevention by T-817 MA. AB - Filamentous tau inclusions are hallmarks of Alzheimer's disease and related neurodegenerative tauopathies, but the molecular mechanisms involved in tau mediated changes in neuronal function and their possible effects on synaptic transmission are unknown. We have evaluated the effects of human tau protein injected directly into the presynaptic terminal axon of the squid giant synapse, which affords functional, structural, and biochemical analysis of its action on the synaptic release process. Indeed, we have found that at physiological concentration recombinant human tau (h-tau42) becomes phosphorylated, produces a rapid synaptic transmission block, and induces the formation of clusters of aggregated synaptic vesicles in the vicinity of the active zone. Presynaptic voltage clamp recordings demonstrate that h-tau42 does not modify the presynaptic calcium current amplitude or kinetics. Analysis of synaptic noise at the post synaptic axon following presynaptic h-tau42 microinjection revealed an initial phase of increase spontaneous transmitter release followed by a marked reduction in noise. Finally, systemic administration of T-817MA, a proposed neuro protective agent, rescued tau-induced synaptic abnormalities. Our results show novel mechanisms of h-tau42 mediated synaptic transmission failure and identify a potential therapeutic agent to treat tau-related neurotoxicity. PMID- 21629768 TI - The neurophysiology of auditory hallucinations - a historical and contemporary review. AB - Electroencephalography and magnetoencephalography are two techniques that distinguish themselves from other neuroimaging methodologies through their ability to directly measure brain-related activity and their high temporal resolution. A large body of research has applied these techniques to study auditory hallucinations. Across a variety of approaches, the left superior temporal cortex is consistently reported to be involved in this symptom. Moreover, there is increasing evidence that a failure in corollary discharge, i.e., a neural signal originating in frontal speech areas that indicates to sensory areas that forthcoming thought is self-generated, may underlie the experience of auditory hallucinations. PMID- 21629769 TI - Characterization of NMDAR-Independent Learning in the Hippocampus. AB - It is currently thought that memory formation requires the activation of NMDA receptors (NMDARs) in the hippocampus. However, recent studies indicate that these receptors are not necessary for all forms of learning. The current experiments examine this issue using context fear conditioning in mice. First, we show that context fear can be acquired without NMDAR activation in previously trained animals. Mice trained in one environment (context A) are subsequently able to learn about a second environment (context B) in the presence of NMDAR antagonists. Second, we demonstrate that NMDAR-independent learning requires the hippocampus and is dependent on protein synthesis. However, unlike NMDAR dependent learning, it is not contingent on the expression of activity-regulated cytoskeleton-associated protein (Arc). Lastly, we present data that suggests NMDAR-independent learning is only observed when recently stimulated neurons are reactivated during conditioning. These data suggest that context fear conditioning modifies synaptic plasticity mechanisms in the hippocampus and allows subsequent learning to occur in the absence of NMDAR activation. PMID- 21629770 TI - Dopamine-signaled reward predictions generated by competitive excitation and inhibition in a spiking neural network model. AB - Dopaminergic neurons in the mammalian substantia nigra display characteristic phasic responses to stimuli which reliably predict the receipt of primary rewards. These responses have been suggested to encode reward prediction-errors similar to those used in reinforcement learning. Here, we propose a model of dopaminergic activity in which prediction-error signals are generated by the joint action of short-latency excitation and long-latency inhibition, in a network undergoing dopaminergic neuromodulation of both spike-timing dependent synaptic plasticity and neuronal excitability. In contrast to previous models, sensitivity to recent events is maintained by the selective modification of specific striatal synapses, efferent to cortical neurons exhibiting stimulus specific, temporally extended activity patterns. Our model shows, in the presence of significant background activity, (i) a shift in dopaminergic response from reward to reward-predicting stimuli, (ii) preservation of a response to unexpected rewards, and (iii) a precisely timed below-baseline dip in activity observed when expected rewards are omitted. PMID- 21629771 TI - Breakdown of the blood-brain barrier during tick-borne encephalitis in mice is not dependent on CD8+ T-cells. AB - Tick-borne encephalitis (TBE) virus causes severe encephalitis with serious sequelae in humans. The disease is characterized by fever and debilitating encephalitis that can progress to chronic illness or fatal infection. In this study, changes in permeability of the blood-brain barrier (BBB) in two susceptible animal models (BALB/c, and C57Bl/6 mice) infected with TBE virus were investigated at various days after infection by measuring fluorescence in brain homogenates after intraperitoneal injection of sodium fluorescein, a compound that is normally excluded from the central nervous system. We demonstrate here that TBE virus infection, in addition to causing fatal encephalitis in mice, induces considerable breakdown of the BBB. The permeability of the BBB increased at later stages of TBE infection when high virus load was present in the brain (i.e., BBB breakdown was not necessary for TBE virus entry into the brain), and at the onset of the first severe clinical symptoms of the disease, which included neurological signs associated with sharp declines in body weight and temperature. The increased BBB permeability was in association with dramatic upregulation of proinflammatory cytokine/chemokine mRNA expression in the brain. Breakdown of the BBB was also observed in mice deficient in CD8(+) T-cells, indicating that these cells are not necessary for the increase in BBB permeability that occurs during TBE. These novel findings are highly relevant to the development of future therapies designed to control this important human infectious disease. PMID- 21629772 TI - Null genotypes of GSTM1 and GSTT1 contribute to risk of cervical neoplasia: an evidence-based meta-analysis. AB - BACKGROUND AND OBJECTIVES: Glutathione S-transferases (GSTs) are multifunctional enzymes that play a key role in the detoxification of varieties of both endogenous products of oxidative stress and exogenous carcinogens. METHODS: In this meta-analysis, twenty-five studies were identified by searching PubMed, EMBASE, ISI Web of Science and CBM databases: 23 evaluated GSTM1 and 19 evaluated GSTT1. Crude odds ratios with corresponding 95% confidence intervals were used to estimate the association between GSTM1 and GSTT1 polymorphisms and risk of cervical neoplasia. Subgroup analyses were conducted by pathological history, ethnicity, source of DNA for genotyping, quality score, and matching variable. RESULTS: The null genotypes of GSTM1 and GSTT1 polymorphisms were associated with a significantly increased risk of cervical neoplasia (for GSTM1: OR = 1.40; 95%CI, 1.19-1.65; for GSTT1: OR = 1.30; 95%CI, 1.05-1.62, respectively). Subgroup analyses showed that the null genotype of GSTM1 increased the risk of cervical neoplasia in Asians, studies with DNA isolation from white blood cells and tissue samples, both high and low quality studies, and matched studies. In GSTM1-GSTT1 interaction analysis, individuals with dual null genotype were associated with a significantly increased risk of cervical neoplasia (OR = 1.72; 95%CI, 1.18-2.51). CONCLUSION: These findings indicate that GSTM1 and GSTT1 polymorphisms, particularly GSTM1-GSTT1 interaction, may play critical roles in the development of cervical neoplasia. A conservative manner should be adopted to interpret these results because of obvious heterogeneity between-study, unadjusted data, and relatively small sample size in this meta-analysis. Well designed studies with larger sample size are of great value to confirm these results. PMID- 21629773 TI - MicroRNA-22 regulates hypoxia signaling in colon cancer cells. AB - MicroRNAs (MiRNAs) are short, non-coding RNA that regulate a variety of cellular functions by suppressing target protein expression. We hypothesized that a set of microRNA regulate tumor responses to hypoxia by inhibiting components of the hypoxia signaling pathway. We found that miR-22 expression in human colon cancer is lower than in normal colon tissue. We also found that miR-22 controls hypoxia inducible factor 1alpha (HIF-1alpha) expression in the HCT116 colon cancer cell line. Over-expression of miR-22 inhibits HIF-1alpha expression, repressing vascular endothelial growth factor (VEGF) production during hypoxia. Conversely, knockdown of endogenous miR-22 enhances hypoxia induced expression of HIF-1alpha and VEGF. The conditioned media from cells over-expressing miR-22 contain less VEGF protein than control cells, and also induce less endothelial cell growth and invasion, suggesting miR-22 in adjacent cells influences endothelial cell function. Taken together, our data suggest that miR-22 might have an anti angiogenic effect in colon cancer. PMID- 21629774 TI - De novo biosynthesis of biodiesel by Escherichia coli in optimized fed-batch cultivation. AB - Biodiesel is a renewable alternative to petroleum diesel fuel that can contribute to carbon dioxide emission reduction and energy supply. Biodiesel is composed of fatty acid alkyl esters, including fatty acid methyl esters (FAMEs) and fatty acid ethyl esters (FAEEs), and is currently produced through the transesterification reaction of methanol (or ethanol) and triacylglycerols (TAGs). TAGs are mainly obtained from oilseed plants and microalgae. A sustainable supply of TAGs is a major bottleneck for current biodiesel production. Here we report the de novo biosynthesis of FAEEs from glucose, which can be derived from lignocellulosic biomass, in genetically engineered Escherichia coli by introduction of the ethanol-producing pathway from Zymomonas mobilis, genetic manipulation to increase the pool of fatty acyl-CoA, and heterologous expression of acyl-coenzyme A: diacylglycerol acyltransferase from Acinetobacter baylyi. An optimized fed-batch microbial fermentation of the modified E. coli strain yielded a titer of 922 mg L(-1) FAEEs that consisted primarily of ethyl palmitate, -oleate, -myristate and -palmitoleate. PMID- 21629775 TI - Temporal anomalies in immunological gene expression in a time series of wild mice: signature of an epidemic? AB - Although the ecological importance of coinfection is increasingly recognized, analyses of microbial pathogen dynamics in wildlife usually focus on an ad hoc subset of the species present due to technological limitations on detection. Here we demonstrate the use of expression profiles for immunological genes (pattern recognition receptors, cytokines and transcription factors) as a means to identify, without preconception, the likelihood of important acute microbial infections in wildlife. Using a wood mouse population in the UK as a model we identified significant temporal clusters of individuals with extreme expression of immunological mediators across multiple loci, typical of an acute microbial infection. These clusters were circumstantially associated with demographic perturbation in the summertime wood mouse population. Animals in one cluster also had significantly higher individual macroparasite burdens than contemporaries with "normal" expression patterns. If the extreme transcriptional profiles observed are induced by an infectious agent then this implicates macroparasites as a possible player in mediating individual susceptibility or resilience to infection. The form of survey described here, combined with next generation nucleic acids sequencing methods for the broad detection of microbial infectious agents in individuals with anomalous immunological transcriptional profiles, could be a powerful tool for revealing unrecognized, ecologically important infectious agents circulating in wildlife populations. PMID- 21629776 TI - The rat IgGFcgammaBP and Muc2 C-terminal domains and TFF3 in two intestinal mucus layers bind together by covalent interaction. AB - BACKGROUND: The secreted proteins from goblet cells compose the intestinal mucus. The aims of this study were to determine how they exist in two intestinal mucus layers. METHODOLOGY/PRINCIPAL FINDINGS: The intestinal mucosa was fixed with Carnoy solution and immunostained. Mucus from the loose layer, the firm layer was gently suctioned or scraped, respectively, lysed in SDS sample buffer with or without DTT, then subjected to the western blotting of rTFF3, rIgGFcgammaBP or rMuc2. The non-reduced or reduced soluble mucus samples in RIPA buffer were co immunoprecipitated to investigate their possible interactions. Polyclonal antibodies for rTFF3, the rIgGFcgammaBP C-terminal domain and the rMuc2 C terminal domain confirmed their localization in the mucus layer and in the mucus collected from the rat intestinal loose layer or firm layer in both western blot and immunoprecipitation experiments. A complex of rTFF3, which was approximately 250 kDa, and a monomer of 6 kDa were present in both layers of the intestinal mucus; rIgGFcgammaBP was present in the complex (250-280 kDa) under non-reducing conditions, but shifted to 164 kDa under reducing conditions in both of the layers. rMuc2 was found mainly in a complex of 214-270 kDa under non-reducing conditions, but it shifted to 140 kDa under reducing conditions. The co immunoprecipitation experiments showed that binding occurs among rTFF3, rIgGFcgammaBP and rMuc2 in the RIPA buffer soluble intestinal mucus. Blocking the covalent interaction by 100 mM DTT in the RIPA buffer soluble intestinal mucus disassociated their binding. CONCLUSIONS/SIGNIFICANCE: Rat goblet cell-secreted TFF3, IgGFcgammaBP and Muc2, existing in the two intestinal mucus layers, are bound together by covalent interactions in the soluble fraction of intestinal mucus and form heteropolymers to be one of the biochemical mechanisms of composing the net-like structure of mucus. PMID- 21629777 TI - McMaster-Toronto Arthritis Patient Preference Disability Questionnaire sensitivity to change in low back pain: influence of shifts in priorities. AB - OBJECTIVE: To assess the sensitivity to change of the McMaster Toronto Arthritis Patient Preference Disability Questionnaire (MACTAR) in chronic low back pain (CLBP) and shifts in patients' priorities of disabling activities over time. METHODS: A prospective longitudinal survey of 100 patients (38 males) with CLBP in a tertiary care teaching hospital. Evaluation at baseline and 6 months by the MACTAR, Quebec Back Pain Disability Questionnaire (QUEBEC), Hospital Anxiety and Depression scale (HAD), Fear-Avoidance Beliefs Questionnaire (FABQ), Coping Strategies Questionnaire (CSQ), and pain and handicap visual analogue scales (VASs). Patients' perceived improvement or worsening of condition was assessed at 6 months. Effect size (ES) and Standardized response mean (SRM) and effect size (ES) were used to evaluate sensitivity to change of the MACTAR. RESULTS: The MACTAR SRM and ES values (SRM = 0.25; ES = 0.37) were among the highest for the instruments evaluated. For patients considering their condition as improved, the SRM was 0.66 and the ES 1. The 3 disability domains, classified by the International Classification of Functioning, Disability and Health (ICF), most often cited as priorities at baseline remained the most cited at follow-up: mobility (40.9% of patients); community, social and civic life (22.7%); and domestic life (22.4%). At 6 months, 48 patients shifted their priorities, for a decrease in MACTAR SRM and ES values for patients considering their condition improved and an increase in these values for those considering their condition deteriorated. CONCLUSIONS: Although the MACTAR has similar sensitivity to change as other outcome measures widely used in CLBP, shifts in patient priorities over time are common and influence scores and sensitivity to change. PMID- 21629778 TI - Propagation of respiratory aerosols by the vuvuzela. AB - Vuvuzelas, the plastic blowing horns used by sports fans, recently achieved international recognition during the FIFA World Cup soccer tournament in South Africa. We hypothesised that vuvuzelas might facilitate the generation and dissemination of respiratory aerosols. To investigate the quantity and size of aerosols emitted when the instrument is played, eight healthy volunteers were asked to blow a vuvuzela. For each individual the concentration of particles in expelled air was measured using a six channel laser particle counter and the duration of blowing and velocity of air leaving the vuvuzela were recorded. To allow comparison with other activities undertaken at sports events each individual was also asked to shout and the measurements were repeated while using a paper cone to confine the exhaled air. Triplicate measurements were taken for each individual. The mean peak particle counts were 658 * 10(3) per litre for the vuvuzela and 3.7 * 10(3) per litre for shouting, representing a mean log(10) difference of 2.20 (95% CI: 2.03,2.36; p < 0.001). The majority (>97%) of particles captured from either the vuvuzela or shouting were between 0.5 and 5 microns in diameter. Mean peak airflows recorded for the vuvuzela and shouting were 6.1 and 1.8 litres per second respectively. We conclude that plastic blowing horns (vuvuzelas) have the capacity to propel extremely large numbers of aerosols into the atmosphere of a size able to penetrate the lower lung. Some respiratory pathogens are spread via contaminated aerosols emitted by infected persons. Further investigation is required to assess the potential of the vuvuzela to contribute to the transmission of aerosol borne diseases. We recommend, as a precautionary measure, that people with respiratory infections should be advised not to blow their vuvuzela in enclosed spaces and where there is a risk of infecting others. PMID- 21629779 TI - Rab11 is required for epithelial cell viability, terminal differentiation, and suppression of tumor-like growth in the Drosophila egg chamber. AB - BACKGROUND: The Drosophila egg chamber provides an excellent system in which to study the specification and differentiation of epithelial cell fates because all of the steps, starting with the division of the corresponding stem cells, called follicle stem cells, have been well described and occur many times over in a single ovary. METHODOLOGY/PRINCIPAL FINDINGS: Here we investigate the role of the small Rab11 GTPase in follicle stem cells (FSCs) and in their differentiating daughters, which include main body epithelial cells, stalk cells and polar cells. We show that rab11-null FSCs maintain their ability to self renew, even though previous studies have shown that FSC self renewal is dependent on maintenance of E-cadherin-based intercellular junctions, which in many cell types, including Drosophila germline stem cells, requires Rab11. We also show that rab11-null FSCs give rise to normal numbers of cells that enter polar, stalk, and epithelial cell differentiation pathways, but that none of the cells complete their differentiation programs and that the epithelial cells undergo premature programmed cell death. Finally we show, through the induction of rab11-null clones at later points in the differentiation program, that Rab11 suppresses tumor-like growth of epithelial cells. Thus, rab11-null epithelial cells arrest differentiation early, assume an aberrant cell morphology, delaminate from the epithelium, and invade the neighboring germline cyst. These phenotypes are associated with defects in E-cadherin localization and a general loss of cell polarity. CONCLUSIONS/SIGNIFICANCE: While previous studies have revealed tumor suppressor or tumor suppressor-like activity for regulators of endocytosis, our study is the first to identify such activity for regulators of endocytic recycling. Our studies also support the recently emerging view that distinct mechanisms regulate junction stability and plasticity in different tissues. PMID- 21629780 TI - Thrombosis is reduced by inhibition of COX-1, but unaffected by inhibition of COX 2, in an acute model of platelet activation in the mouse. AB - BACKGROUND: Clinical use of selective inhibitors of cyclooxygenase (COX)-2 appears associated with increased risk of thrombotic events. This is often hypothesised to reflect reduction in anti-thrombotic prostanoids, notably PGI(2), formed by COX-2 present within endothelial cells. However, whether COX-2 is actually expressed to any significant extent within endothelial cells is controversial. Here we have tested the effects of acute inhibition of COX on platelet reactivity using a functional in vivo approach in mice. METHODOLOGY/PRINCIPAL FINDINGS: A non-lethal model of platelet-driven thromboembolism in the mouse was used to assess the effects of aspirin (7 days orally as control) diclofenac (1 mg.kg(-1), i.v.) and parecoxib (0.5 mg.kg(-1), i.v.) on thrombus formation induced by collagen or the thromboxane (TX) A(2) mimetic, U46619. The COX inhibitory profiles of the drugs were confirmed in mouse tissues ex vivo. Collagen and U46619 caused in vivo thrombus formation with the former, but not latter, sensitive to oral dosing with aspirin. Diclofenac inhibited COX-1 and COX-2 ex vivo and reduced thrombus formation in response to collagen, but not U46619. Parecoxib inhibited only COX-2 and had no effect upon thrombus formation caused by either agonist. CONCLUSIONS/SIGNIFICANCE: Inhibition of COX-1 by diclofenac or aspirin reduced thrombus formation induced by collagen, which is partly dependent upon platelet-derived TXA(2), but not that induced by U46619, which is independent of platelet TXA(2). These results are consistent with the model demonstrating the effects of COX-1 inhibition in platelets, but provide no support for the hypothesis that acute inhibition of COX-2 in the circulation increases thrombosis. PMID- 21629781 TI - Range-expanding populations of a globally introduced weed experience negative plant-soil feedbacks. AB - BACKGROUND: Biological invasions are fundamentally biogeographic processes that occur over large spatial scales. Interactions with soil microbes can have strong impacts on plant invasions, but how these interactions vary among areas where introduced species are highly invasive vs. naturalized is still unknown. In this study, we examined biogeographic variation in plant-soil microbe interactions of a globally invasive weed, Centaurea solstitialis (yellow starthistle). We addressed the following questions (1) Is Centaurea released from natural enemy pressure from soil microbes in introduced regions? and (2) Is variation in plant soil feedbacks associated with variation in Centaurea's invasive success? METHODOLOGY/PRINCIPAL FINDINGS: We conducted greenhouse experiments using soils and seeds collected from native Eurasian populations and introduced populations spanning North and South America where Centaurea is highly invasive and noninvasive. Soil microbes had pervasive negative effects in all regions, although the magnitude of their effect varied among regions. These patterns were not unequivocally congruent with the enemy release hypothesis. Surprisingly, we also found that Centaurea generated strong negative feedbacks in regions where it is the most invasive, while it generated neutral plant-soil feedbacks where it is noninvasive. CONCLUSIONS/SIGNIFICANCE: Recent studies have found reduced below ground enemy attack and more positive plant-soil feedbacks in range-expanding plant populations, but we found increased negative effects of soil microbes in range-expanding Centaurea populations. While such negative feedbacks may limit the long-term persistence of invasive plants, such feedbacks may also contribute to the success of invasions, either by having disproportionately negative impacts on competing species, or by yielding relatively better growth in uncolonized areas that would encourage lateral spread. Enemy release from soil-borne pathogens is not sufficient to explain the success of this weed in such different regions. The biogeographic variation in soil-microbe effects indicates that different mechanisms may operate on this species in different regions, thus establishing geographic mosaics of species interactions that contribute to variation in invasion success. PMID- 21629782 TI - Autoantibodies against a 43 KDa muscle protein in inclusion body myositis. AB - BACKGROUND: Inclusion body myositis (IBM) is a poorly understood and refractory autoimmune muscle disease. Though widely believed to have no significant humoral autoimmunity, we sought to identify novel autoantibodies with high specificity for this disease. METHODOLOGY/PRINCIPAL FINDINGS: Plasma autoantibodies from 65 people, including 25 with IBM, were analyzed by immunoblots against normal human muscle. Thirteen of 25 (52%) IBM patient samples recognized an approximately 43 kDa muscle protein. No other disease (N = 25) or healthy volunteer (N = 15) samples recognized this protein. CONCLUSIONS: Circulating antibodies against a 43 kDa muscle autoantigen may lead to the discovery of a novel biomarker for IBM. Its high specificity for IBM among patients with autoimmune myopathies furthermore suggests a relationship to disease pathogenesis. PMID- 21629783 TI - Changing patterns of malaria epidemiology between 2002 and 2010 in Western Kenya: the fall and rise of malaria. AB - BACKGROUND: The impact of insecticide treated nets (ITNs) on reducing malaria incidence is shown mainly through data collection from health facilities. Routine evaluation of long-term epidemiological and entomological dynamics is currently unavailable. In Kenya, new policies supporting the provision of free ITNs were implemented nationwide in June 2006. To evaluate the impacts of ITNs on malaria transmission, we conducted monthly surveys in three sentinel sites with different transmission intensities in western Kenya from 2002 to 2010. METHODS AND FINDINGS: Longitudinal samplings of malaria parasite prevalence in asymptomatic school children and vector abundance in randomly selected houses were undertaken monthly from February 2002. ITN ownership and usage surveys were conducted annually from 2004 to 2010. Asymptomatic malaria parasite prevalence and vector abundances gradually decreased in all three sites from 2002 to 2006, and parasite prevalence reached its lowest level from late 2006 to early 2007. The abundance of the major malaria vectors, Anopheles funestus and An. gambiae, increased about 5-10 folds in all study sites after 2007. However, the resurgence of vectors was highly variable between sites and species. By 2010, asymptomatic parasite prevalence in Kombewa had resurged to levels recorded in 2004/2005, but the resurgence was smaller in magnitude in the other sites. Household ITN ownership was at 50-70% in 2009, but the functional and effective bed net coverage in the population was estimated at 40.3%, 49.4% and 28.2% in 2010 in Iguhu, Kombewa, and Marani, respectively. CONCLUSION: The resurgence in parasite prevalence and malaria vectors has been observed in two out of three sentinel sites in western Kenya despite a high ownership of ITNs. The likely factors contributing to malaria resurgence include reduced efficacy of ITNs, insecticide resistance in mosquitoes and lack of proper use of ITNs. These factors should be targeted to avoid further resurgence of malaria transmission. PMID- 21629784 TI - A tri-marker proliferation index predicts biochemical recurrence after surgery for prostate cancer. AB - Prostate cancer exhibits tremendous variability in clinical behavior, ranging from indolent to lethal disease. Better prognostic markers are needed to stratify patients for appropriately aggressive therapy. By expression profiling, we can identify a proliferation signature variably expressed in prostate cancers. Here, we asked whether one or more tissue biomarkers might capture that information, and provide prognostic utility. We assayed three proliferation signature genes: MKI67 (Ki-67; also a classic proliferation biomarker), TOP2A (DNA topoisomerase II, alpha), and E2F1 (E2F transcription factor 1). Immunohistochemical staining was evaluable on 139 radical prostatectomy cases (in tissue microarray format), with a median clinical follow-up of eight years. Each of the three proliferation markers was by itself prognostic. Notably, combining the three markers together as a "proliferation index" (0 or 1, vs. 2 or 3 positive markers) provided superior prognostic performance (hazard ratio = 2.6 (95% CI: 1.4-4.9); P = 0.001). In a multivariate analysis that included preoperative serum prostate specific antigen (PSA) levels, Gleason grade and pathologic tumor stage, the composite proliferation index remained a significant predictor (P = 0.005). Analysis of receiver-operating characteristic (ROC) curves confirmed the improved prognostication afforded by incorporating the proliferation index (compared to the clinicopathologic data alone). Our findings highlight the potential value of a multi-gene signature-based diagnostic, and define a tri-marker proliferation index with possible utility for improved prognostication and treatment stratification in prostate cancer. PMID- 21629785 TI - Lack of the receptor for advanced glycation end-products attenuates E. coli pneumonia in mice. AB - BACKGROUND: The receptor for advanced glycation end-products (RAGE) has been suggested to modulate lung injury in models of acute pulmonary inflammation. To study this further, model systems utilizing wild type and RAGE knockout (KO) mice were used to determine the role of RAGE signaling in lipopolysaccharide (LPS) and E. coli induced acute pulmonary inflammation. The effect of intraperitoneal (i.p.) and intratracheal (i.t.) administration of mouse soluble RAGE on E. coli injury was also investigated. METHODOLOGY/PRINCIPAL FINDINGS: C57BL/6 wild type and RAGE KO mice received an i.t. instillation of LPS, E. coli, or vehicle control. Some groups also received i.p. or i.t. administration of mouse soluble RAGE. After 24 hours, the role of RAGE expression on inflammation was assessed by comparing responses in wild type and RAGE KO. RAGE protein levels decreased in wild type lung homogenates after treatment with either LPS or bacteria. In addition, soluble RAGE and HMGB1 increased in the BALF after E. coli instillation. RAGE KO mice challenged with LPS had the same degree of inflammation as wild type mice. However, when challenged with E. coli, RAGE KO mice had significantly less inflammation when compared to wild type mice. Most cytokine levels were lower in the BALF of RAGE KO mice compared to wild type mice after E. coli injury, while only monocyte chemotactic protein-1, MCP-1, was lower after LPS challenge. Neither i.p. nor i.t. administration of mouse soluble RAGE attenuated the severity of E. coli injury in wild type mice. CONCLUSIONS/SIGNIFICANCE: Lack of RAGE in the lung does not protect against LPS induced acute pulmonary inflammation, but attenuates injury following live E. coli challenge. These findings suggest that RAGE mediates responses to E. coli associated pathogen-associated molecular pattern molecules other than LPS or other bacterial specific signaling responses. Soluble RAGE treatment had no effect on inflammation. PMID- 21629786 TI - Diosgenin, a steroidal saponin, inhibits migration and invasion of human prostate cancer PC-3 cells by reducing matrix metalloproteinases expression. AB - BACKGROUND: Diosgenin, a steroidal saponin obtained from fenugreek (Trigonella foenum graecum), was found to exert anti-carcinogenic properties, such as inhibiting proliferation and inducing apoptosis in a variety of tumor cells. However, the effect of diosgenin on cancer metastasis remains unclear. The aim of the study is to examine the effect of diosgenin on migration and invasion in human prostate cancer PC-3 cells. METHODS AND PRINCIPAL FINDINGS: Diosgenin inhibited proliferation of PC-3 cells in a dose-dependent manner. When treated with non-toxic doses of diosgenin, cell migration and invasion were markedly suppressed by in vitro wound healing assay and Boyden chamber invasion assay, respectively. Furthermore, diosgenin reduced the activities of matrix metalloproteinase-2 (MMP-2) and MMP-9 by gelatin zymography assay. The mRNA level of MMP-2, -9, -7 and extracellular inducer of matrix metalloproteinase (EMMPRIN) were also suppressed while tissue inhibitor of metalloproteinase-2 (TIMP-2) was increased by diosgenin. In addition, diosgenin abolished the expression of vascular endothelial growth factor (VEGF) in PC-3 cells and tube formation of endothelial cells. Our immunoblotting assays indicated that diosgenin potently suppressed the phosphorylation of phosphatidylinositide-3 kinase (PI3K), Akt, extracellular signal regulating kinase (ERK) and c-Jun N-terminal kinase (JNK). In addition, diosgenin significantly decreased the nuclear level of nuclear factor kappa B (NF-kappaB), suggesting that diosgenin inhibited NF-kappaB activity. CONCLUSION/SIGNIFICANCE: The results suggested that diosgenin inhibited migration and invasion of PC-3 cells by reducing MMPs expression. It also inhibited ERK, JNK and PI3K/Akt signaling pathways as well as NF-kappaB activity. These findings reveal new therapeutic potential for diosgenin in anti-metastatic therapy. PMID- 21629787 TI - Acquired type III secretion system determines environmental fitness of epidemic Vibrio parahaemolyticus in the interaction with bacterivorous protists. AB - Genome analyses of marine microbial communities have revealed the widespread occurrence of genomic islands (GIs), many of which encode for protein secretion machineries described in the context of bacteria-eukaryote interactions. Yet experimental support for the specific roles of such GIs in aquatic community interactions remains scarce. Here, we test for the contribution of type III secretion systems (T3SS) to the environmental fitness of epidemic Vibrio parahaemolyticus. Comparisons of V. parahaemolyticus wild types and T3SS defective mutants demonstrate that the T3SS encoded on genome island VPaI-7 (T3SS 2) promotes survival of V. parahaemolyticus in the interaction with diverse protist taxa. Enhanced persistence was found to be due to T3SS-2 mediated cytotoxicity and facultative parasitism of V. parahaemolyticus on coexisting protists. Growth in the presence of bacterivorous protists and the T3SS-2 genotype showed a strong correlation across environmental and clinical isolates of V. parahaemolyticus. Short-term microcosm experiments provide evidence that protistan hosts facilitate the invasion of T3SS-2 positive V. parahaemolyticus into a coastal plankton community, and that water temperature and productivity further promote enhanced survival of T3SS-2 positive V. parahaemolyticus. This study is the first to describe the fitness advantage of GI-encoded functions in a microbial food web, which may provide a mechanistic explanation for the global spread and the seasonal dynamics of V. parahaemolyticus pathotypes, including the pandemic serotype cluster O3:K6, in aquatic environments. PMID- 21629788 TI - Application of two-part statistics for comparison of sequence variant counts. AB - Investigation of microbial communities, particularly human associated communities, is significantly enhanced by the vast amounts of sequence data produced by high throughput sequencing technologies. However, these data create high-dimensional complex data sets that consist of a large proportion of zeros, non-negative skewed counts, and frequently, limited number of samples. These features distinguish sequence data from other forms of high-dimensional data, and are not adequately addressed by statistical approaches in common use. Ultimately, medical studies may identify targeted interventions or treatments, but lack of analytic tools for feature selection and identification of taxa responsible for differences between groups, is hindering advancement. The objective of this paper is to examine the application of a two-part statistic to identify taxa that differ between two groups. The advantages of the two-part statistic over common statistical tests applied to sequence count datasets are discussed. Results from the t-test, the Wilcoxon test, and the two-part test are compared using sequence counts from microbial ecology studies in cystic fibrosis and from cenote samples. We show superior performance of the two-part statistic for analysis of sequence data. The improved performance in microbial ecology studies was independent of study type and sequence technology used. PMID- 21629789 TI - Mouse transgenesis identifies conserved functional enhancers and cis-regulatory motif in the vertebrate LIM homeobox gene Lhx2 locus. AB - The vertebrate Lhx2 is a member of the LIM homeobox family of transcription factors. It is essential for the normal development of the forebrain, eye, olfactory system and liver as well for the differentiation of lymphoid cells. However, despite the highly restricted spatio-temporal expression pattern of Lhx2, nothing is known about its transcriptional regulation. In mammals and chicken, Crb2, Dennd1a and Lhx2 constitute a conserved linkage block, while the intervening Dennd1a is lost in the fugu Lhx2 locus. To identify functional enhancers of Lhx2, we predicted conserved noncoding elements (CNEs) in the human, mouse and fugu Crb2-Lhx2 loci and assayed their function in transgenic mouse at E11.5. Four of the eight CNE constructs tested functioned as tissue-specific enhancers in specific regions of the central nervous system and the dorsal root ganglia (DRG), recapitulating partial and overlapping expression patterns of Lhx2 and Crb2 genes. There was considerable overlap in the expression domains of the CNEs, which suggests that the CNEs are either redundant enhancers or regulating different genes in the locus. Using a large set of CNEs (810 CNEs) associated with transcription factor-encoding genes that express predominantly in the central nervous system, we predicted four over-represented 8-mer motifs that are likely to be associated with expression in the central nervous system. Mutation of one of them in a CNE that drove reporter expression in the neural tube and DRG abolished expression in both domains indicating that this motif is essential for expression in these domains. The failure of the four functional enhancers to recapitulate the complete expression pattern of Lhx2 at E11.5 indicates that there must be other Lhx2 enhancers that are either located outside the region investigated or divergent in mammals and fishes. Other approaches such as sequence comparison between multiple mammals are required to identify and characterize such enhancers. PMID- 21629790 TI - Localization of the cochlear amplifier in living sensitive ears. AB - BACKGROUND: To detect soft sounds, the mammalian cochlea increases its sensitivity by amplifying incoming sounds up to one thousand times. Although the cochlear amplifier is thought to be a local cellular process at an area basal to the response peak on the spiral basilar membrane, its location has not been demonstrated experimentally. METHODOLOGY AND PRINCIPAL FINDINGS: Using a sensitive laser interferometer to measure sub-nanometer vibrations at two locations along the basilar membrane in sensitive gerbil cochleae, here we show that the cochlea can boost soft sound-induced vibrations as much as 50 dB/mm at an area proximal to the response peak on the basilar membrane. The observed amplification works maximally at low sound levels and at frequencies immediately below the peak-response frequency of the measured apical location. The amplification decreases more than 65 dB/mm as sound levels increases. CONCLUSIONS AND SIGNIFICANCE: We conclude that the cochlea amplifier resides at a small longitudinal region basal to the response peak in the sensitive cochlea. These data provides critical information for advancing our knowledge on cochlear mechanisms responsible for the remarkable hearing sensitivity, frequency selectivity and dynamic range. PMID- 21629791 TI - Bistable percepts in the brain: FMRI contrasts monocular pattern rivalry and binocular rivalry. AB - The neural correlates of binocular rivalry have been actively debated in recent years, and are of considerable interest as they may shed light on mechanisms of conscious awareness. In a related phenomenon, monocular rivalry, a composite image is shown to both eyes. The subject experiences perceptual alternations in which the two stimulus components alternate in clarity or salience. The experience is similar to perceptual alternations in binocular rivalry, although the reduction in visibility of the suppressed component is greater for binocular rivalry, especially at higher stimulus contrasts. We used fMRI at 3T to image activity in visual cortex while subjects perceived either monocular or binocular rivalry, or a matched non-rivalrous control condition. The stimulus patterns were left/right oblique gratings with the luminance contrast set at 9%, 18% or 36%. Compared to a blank screen, both binocular and monocular rivalry showed a U shaped function of activation as a function of stimulus contrast, i.e. higher activity for most areas at 9% and 36%. The sites of cortical activation for monocular rivalry included occipital pole (V1, V2, V3), ventral temporal, and superior parietal cortex. The additional areas for binocular rivalry included lateral occipital regions, as well as inferior parietal cortex close to the temporoparietal junction (TPJ). In particular, higher-tier areas MT+ and V3A were more active for binocular than monocular rivalry for all contrasts. In comparison, activation in V2 and V3 was reduced for binocular compared to monocular rivalry at the higher contrasts that evoked stronger binocular perceptual suppression, indicating that the effects of suppression are not limited to interocular suppression in V1. PMID- 21629792 TI - C1 Domain-targeted isophthalate derivatives induce cell elongation and cell cycle arrest in HeLa cells. AB - Diacylglycerol (DAG)-mediated signaling pathways, such as those mediated by protein kinase C (PKC), are central in regulating cell proliferation and apoptosis. DAG-responsive C1 domains are therefore considered attractive drug targets. Our group has designed a novel class of compounds targeted to the DAG binding site within the C1 domain of PKC. We have previously shown that these 5 (hydroxymethyl)isophthalates modulate PKC activation in living cells. In this study we investigated their effects on HeLa human cervical cancer cell viability and proliferation by using standard cytotoxicity tests and an automated imaging platform with machine vision technology. Cellular effects and their mechanisms were further characterized with the most potent compound, HMI-1a3. Isophthalate derivatives with high affinity to the PKC C1 domain exhibited antiproliferative and non-necrotic cytotoxic effects on HeLa cells. The anti-proliferative effect was irreversible and accompanied by cell elongation. HMI-1a3 induced down regulation of retinoblastoma protein and cyclins A, B1, D1, and E. Effects of isophthalates on cell morphology, cell proliferation and expression of cell cycle related proteins were different from those induced by phorbol 12-myristate-13 acetate (PMA) or bryostatin 1, but correlated closely to binding affinities. Therefore, the results strongly indicate that the effect is C1 domain-mediated. PMID- 21629793 TI - Concentration independent modulation of local micromechanics in a fibrin gel. AB - Methods for tuning extracellular matrix (ECM) mechanics in 3D cell culture that rely on increasing the concentration of either protein or cross-linking molecules fail to control important parameters such as pore size, ligand density, and molecular diffusivity. Alternatively, ECM stiffness can be modulated independently from protein concentration by mechanically loading the ECM. We have developed a novel device for generating stiffness gradients in naturally derived ECMs, where stiffness is tuned by inducing strain, while local mechanical properties are directly determined by laser tweezers based active microrheology (AMR). Hydrogel substrates polymerized within 35 mm diameter Petri dishes are strained non-uniformly by the precise rotation of an embedded cylindrical post, and exhibit a position-dependent stiffness with little to no modulation of local mesh geometry. Here we present the device in the context of fibrin hydrogels. First AMR is used to directly measure local micromechanics in unstrained hydrogels of increasing fibrin concentration. Changes in stiffness are then mapped within our device, where fibrin concentration is held constant. Fluorescence confocal imaging and orbital particle tracking are used to quantify structural changes in fibrin on the micro and nano levels respectively. The micromechanical strain stiffening measured by microrheology is not accompanied by ECM microstructural changes under our applied loads, as measured by confocal microscopy. However, super-resolution orbital tracking reveals nanostructural straightening, lengthening, and reduced movement of fibrin fibers. Furthermore, we show that aortic smooth muscle cells cultured within our device are morphologically sensitive to the induced mechanical gradient. Our results demonstrate a powerful cell culture tool that can be used in the study of mechanical effects on cellular physiology in naturally derived 3D ECM tissues. PMID- 21629794 TI - Internal Mammary Arterial Injury from Lead Extraction: A Clinically Subtle yet Important Complication of Implantable Device Removal. AB - Percutaneous implantable device extraction has increased in recent years and is associated with small but significant risk. Arteriovenous fistula formation is an uncommon complication of this procedure. We report two cases where lead extraction was complicated by an arteriovenous fistula between the left internal mammary artery and the left brachiocephalic vein. In both cases, the patients were asymptomatic and the presence of a continuous murmur in the left subclavicular region led to the appropriate diagnosis. These were successfully treated with coil embolization. Auscultation around prior extraction sites should be routinely done to aid in the diagnosis of this potentially harmful complication. PMID- 21629795 TI - The value of cryosurgery in treating a case of thoracic chondrosarcoma. AB - Chondrosarcomas of the spine are rare and difficult to treat. In this paper a case of thoracic chondrosarcoma is presented. Chondrosarcomas of the spine are generally smaller, more difficult to excise and are followed by higher local recurrence compared with chondrosarcomas of the peripheral skeleton. The tumor is radio- and chemoresistant, making the surgical treatment of utmost importance. The most important prognostic factor for local control is wide or marginal tumor resection. Our patient was treated in two stages, with total excision of the tumor, using cryosurgery. Liquid nitrogen was used to freeze the damaged tissue at a cellular level and made the excision more efficient. PMID- 21629796 TI - Mediterranean diet mediates the adverse effect of depressive symptomatology on short-term outcome in elderly survivors from an acute coronary event. AB - Aims. We evaluated the interaction effect between depressive symptoms and dietary habits on 30-day development of cardiovascular disease (CVD) (death or rehospitalization) in elderly, acute coronary syndrome (ACS) survivors. Methods. During 2006-2008, we recorded 277 nonfatal, consecutive ACS admissions (75 +/- 6 years, 70% males, 70% had diagnosis of myocardial infarction) with complete 30 day follow-up. Assessment of recent depressive symptoms was based on the CES-D scale. Among sociodemographic, bioclinical, lifestyle characteristics, the MedDietScore that assesses the inherent characteristics of the Mediterranean diet was applied. Results. 22% of the ACS pts developed a CVD event during the first 30 days (14.8% rehospitalization and 9.4% death). Patients in the upper tertile of the CES-D scale (i.e., >18) had higher incidence of CVD events as compared with those in the lowest tertile (21% versus 8%, P = .01). Multiple logistic regression analysis revealed that 1-unit increase in CES-D was associated with 4% higher odds (95% CI 1.008-1.076, P = .01) of CVD events; however, when MedDietScore was entered in the model, CES-D lost its significance (P = .20). Conclusion. Short-term depressive symptoms are related to a worsen 30-day prognosis of ACS patients; however, this relationship was mediated by Mediterranean diet adherence. PMID- 21629797 TI - Systemic lupus erythematosus presenting as thrombotic thrombocytopenia purpura: how close is close enough? AB - Thrombotic thrombocytopenic purpura (TTP) is an uncommon life-threatening disease characterized by microangiopathic hemolytic anemia and thrombocytopenia, commonly associated with infections, malignancy, drugs, and autoimmune diseases. We report a case of 19-year-old previously healthy female that presents with anemia and thrombocytopenia diagnosed with thrombotic thrombocytopenic purpura that was treated successfully with plasmapheresis and corticosteroids. Laboratory findings also revealed antinuclear antibodies and antibodies to double-stranded DNA. Two weeks after presentation developed inflammatory arthritis, fulfilling diagnostic criteria for systemic lupus erythematosus (SLE). Prompt diagnosis and treatment with plasma exchange and corticosteroids should be instituted as soon as the diagnosis of TTP is suspected, even if other diagnoses, including lupus, are possible. When present, the coexistence of these two etiologies can have a higher mortality than either disease alone. An underlying diagnosis of SLE should be considered in all patients presenting TTP and the study of this association may provide a better understanding of their immune-mediated pathophysiology. PMID- 21629798 TI - Vascular endothelial growth factor inhibitor-induced hypertension: basics for primary care providers. AB - Frequently, primary care providers continue to manage the overall medical care of cancer patients. With newer and often more potent antitumor agents, patients may present to their local physicians with drug-induced toxicities such as hypertension induced by vascular endothelial growth factor (VEGF) inhibitors. It is imperative that these healthcare providers are aware of basic aspects of this drug class, as its use has increased significantly in the last several years. Uncontrolled or malignant hypertension due to these agents should be recognized readily and treated early to prevent more severe outcomes. This overview provides a brief background on the role of VEGF and angiogenesis in tumor metabolism as well as theories of the mechanism of VEGF inhibitors and hypertension. Helpful clinical practice aspects including the types of inhibitors used in the United States and their pharmacologic characteristics will be discussed. Also, diagnosis and treatment of hypertension induced by vascular endothelial growth factors are reviewed. A summary of key aspects of this drug class and hypertension is included. PMID- 21629799 TI - Prolonged Refractory Hypotension following Combined Amlodipine and Losartan Ingestion Responsive to Metaraminol. AB - Introduction. Overdose with the calcium channel blocker amlodipine can cause profound hypotension that may be exacerbated by the concurrent ingestion of an angiotensin II receptor antagonist. Best management of such overdoses is uncertain although the use of hyperinsulinaemia-euglycaemia (HIE) has been recommended. Case report. We report a case of mixed amlodipine and losartan overdose in a 50-year-old lady. Severe hypotension was resistant to conventional vasopressors and high-dose insulin/euglycaemia, but did respond to a metaraminol infusion. Conclusion. A trial of metaraminol early in severe cases of calcium channel blocker and angiotensin II receptor antagonist toxicity may be of benefit, especially when conventional ionotropic treatment measures are failing. PMID- 21629800 TI - Interactions between CKD and MetS and the Development of CVD. PMID- 21629802 TI - Myasthenia gravis development and crisis subsequent to multiple sclerosis. AB - During the last decade, sporadic combination of multiple sclerosis (MS) and myasthenia gravis (MG) has been reported repeatedly. Although these are anecdotal, they are important enough to raise concerns about co-occurrence of MG and MS. Here, we present a case of an MS patient who developed an MG crisis. She had received interferon for relapsing remitting MS. Interestingly, she developed an MG crisis 4 years after the diagnosis of MS. MS and MG have relatively the same distribution for age, corresponding to the younger peak of the bimodal age distribution in MG. They also share some HLA typing characteristics. Furthermore, some evidences support the role of systemic immune dysregulation due to a genetic susceptibility that is common to these two diseases. The association may be underdiagnosed because of the possible overlap of symptoms especially bulbar manifestations in which either MG or MS can mimic each other, leading to underestimating incidence of the combination. The evidence warrants physicians, especially neurologists, to always consider the possibility of the other disease when encountering any patients either with MS or MG. Anecdotal and sporadic reports of combination of multiple sclerosis (MS) and myasthenia gravis (MG) have been raised concerns about co-occurrence of them. PMID- 21629801 TI - The registry and follow-up of complex pediatric therapies program of Western Canada: a mechanism for service, audit, and research after life-saving therapies for young children. AB - Newly emerging health technologies are being developed to care for children with complex cardiac defects. Neurodevelopmental and childhood school-related outcomes are of great interest to parents of children receiving this care, care providers, and healthcare administrators. Since the 1970s, neonatal follow-up clinics have provided service, audit, and research for preterm infants as care for these at risk children evolved. We have chosen to present for this issue the mechanism for longitudinal follow-up of survivors that we have developed for western Canada patterned after neonatal follow-up. Our program provides registration for young children receiving complex cardiac surgery, heart transplantation, ventricular assist device support, and extracorporeal life support among others. The program includes multidisciplinary assessments with appropriate neurodevelopmental intervention, active quality improvement evaluations, and outcomes research. Through this mechanism, consistently high (96%) follow-up over two years is maintained. PMID- 21629803 TI - A Case of Giant Bladder Carcinosarcoma without Submucosal Invasion. AB - Carcinosarcoma is a rare biphasic neoplasia containing both malignant mesenchymal and epithelial elements. Bladder carcinosarcoma commonly presented as high-grade, advanced stage, and aggressive behavior with a poor prognosis. An 83-year-old male presented with painless gross hematuria to our hospital. Cystoscopy revealed massive nonpapillary bladder tumor on the right wall. The 91 g tumor could be completely removed with transurethral resection. Histology of the tumor was diagnosed as carcinosarcoma with no submucosal invasion composed of biphasic malignant epithelial and mesenchymal cells. Epithelial malignancy was urothelial cancer and mesenchymal one was chondrosarcoma and leiomyosarcoma. The specimens taken at the second-look TUR-Bt revealed that carcinoma in situ (urothelial cancer) but not sarcoma existed at the mucosa surrounding the previous tumor site. 80 mg of BCG instillation intravesically every week for six weeks was successfully administered to the patient. There is no tumor recurrence for 6 months after treatments. PMID- 21629804 TI - Gastric perforation associated with tuberculosis: a case report. AB - Gastric tuberculosis is a rare presentation of tuberculosis infection. Gastric perforation associated with tuberculosis is exceedingly rare with five previously published cases. We present a case of a male patient that developed presumed gastric tuberculosis secondary to pulmonary tuberculosis infection. He subsequently developed gastric perforation and sepsis, for which he was treated both surgically and medically. Despite ongoing antituberculosis treatment, the patient's condition worsened and the patient died secondary to multiorgan failure. This case highlights gastric perforation as a rare but devastating complication of pulmonary tuberculosis. PMID- 21629805 TI - Tophaceous gout and renal insufficiency: a new solution for an old therapeutic dilemma. AB - The prevalence of gout is increasing with increased life expectancy. Approximately half of the patients with gout have some degree of renal impairment. If both conditions persistently coexist, and in severe tophaceous gout, in particular, treatment has been difficult. We here report on the case of an 87-year-old woman, who had been suffering from recurrent gouty arthritis over 4 years. Monthly polyarthritis attacks were accompanied by subcutaneous tophi. Serum uric acid levels were constantly above 600 MUmol/L (10 mg/dL). Allopurinol was no option because of intolerance, while benzbromarone was ineffective because of renal impairment. Therefore, the novel xanthin oxidase inhibitor febuxostat was started, achieving rapid control of serum urate levels (<360 MUmol/L). After initial worsening of inflammation in the first weeks, gouty attacks stopped and all tophi resolved within the following 10 months. Renal function remained stable. PMID- 21629806 TI - Relapse of neuromyelitis optica spectrum disorder associated with intravenous lidocaine. AB - Lidocaine unmasks silent symptoms and eases neuropathic pain in multiple sclerosis patients; however, the effects of lidocaine in neuromyelitis optica have never been reported. We describe the case of a 59-year-old Japanese woman with neuromyelitis optica spectrum disorder who developed optic neuritis 1 day after intravenous lidocaine injection for treating allodynia. Her symptom seemed to result from a relapse of neuromyelitis optica induced by lidocaine administration, and not because of the transient effects of intravenous lidocaine administration. The possibility that lidocaine administration results in relapse of neuromyelitis optica due to its immunomodulating effects cannot be ruled out. PMID- 21629807 TI - Management of an oral ingestion of transdermal fentanyl patches: a case report and literature review. AB - Purpose. Fentanyl is available as a transdermal system for the treatment of chronic pain in opioid-tolerant patients; however, it carries a black box warning due to both the potency of the product and the potential for abuse. In this report, we describe a case of transbuccal and gastrointestinal ingestion of fentanyl patches and the management of such ingestion. Summary. A 32-year-old man was brought to the emergency department (ED) via emergency medical services for toxic ingestion and suicide attempt. The patient chewed and ingested two illegally purchased transdermal fentanyl patches. In the ED, the patient was obtunded, dizzy and drowsy. Initial vital signs showed the patient to be afebrile and normotensive with a heart rate of 63, respiratory rate of 16, and oxygen saturation of 100% on 2 liters nasal cannula after administration of 2 milligrams of intravenous naloxone. The patient was treated with whole bowel irrigation and continuous intravenous naloxone infusion for approximately 48 hours without complications. Conclusion. Despite numerous case reports describing oral ingestion of fentanyl patches, information on the management of such intoxication is lacking. We report successful management of such a case utilizing whole bowel irrigation along with intravenous push and continuous infusion naloxone. PMID- 21629808 TI - Successful treatment of stent knot in the proximal ureter using ureteroscopy and holmium laser. AB - Knotted ureteral stent is rare yet tedious complication that might represent a treatment challenge to the endourologist. Only twelve cases of knotted stent have been reported. Different management options have been reported, including simple traction, ureteroscopy, percutaneous removal, and open surgery. In this paper, we present the successful untying of the knot using ureteroscopy with holmium laser. PMID- 21629809 TI - Toxocariasis presenting as encephalomyelitis. AB - We describe a farmer who presented with a clinical picture of a transverse thoracic myelitis. MRI showed inflammatory lesions in brain and thoracic spinal cord. Toxocariasis was suspected because of eosinophilia in blood and cerebrospinal fluid, and this diagnosis was confirmed immunologically. He was successfully treated with antihelminthics in combination with corticosteroids. Neurotoxocariasis is rare and diagnosis can be difficult because of the different and atypical clinical manifestations. It should be considered in every case of central neurological syndrome associated with eosinophilia. PMID- 21629810 TI - Vulvar merkel carcinoma: a case report. AB - This is a new case of Merkel cell carcinoma of the vulva. It is a rare neuroendocrine carcinoma with an aggressive behavior. Because of its rarity in this location, it is not clear whether it behaves differently from the usual neuroendocrine carcinomas of the skin. A case of a 63-year-old patient with vulvar Merkel carcinoma is presented. The clinical presentation, microscopic and immunohistochemical features, and treatment are discussed. PMID- 21629811 TI - Pheochromocytoma presenting as acute heart failure leading to cardiogenic shock and multiorgan failure. AB - Pheochromocytoma is an endocrine tumor classically presenting with headache, paroxysmal hypertension, and palpitations. We discuss the case of a young male, presenting with acute heart failure and cardiogenic shock requiring stabilization with an intra-aortic balloon pump and a combination of ionotropes and vasopressors. Pheochromocytoma was diagnosed by CT scan, as well as urine and plasma metanephrines. After pretreatment with phenoxy-benzamine, the patient underwent adrenalectomy with subsequent cardiovascular stabilization and full recovery. Unfortunately, pheochromocytoma often remains undiagnosed. Given the ample diagnostic tools and good prognosis when treated suitably, the diagnosis should be entertained early in patients, presenting with unexplained cardiovascular compromise. PMID- 21629812 TI - Anterior uveitis as an initial manifestation of polymyalgia rheumatica. AB - A 74-year-old woman without contributory medical history presented with acute iridocyclitis in the right eye. Although the iridocyclitis disappeared within two weeks under topical steroid, she complained of acute progressing bilateral shoulder pain and morning stiffness of upper extremities. She was diagnosed as having polymyalgia rheumatica (PMR), and iridocyclitis was considered as its related manifestation. PMR and giant cell arteritis (GCA) are closely related conditions and frequently occur together. GCA with uveitis has been rarely noted. However, ocular symptoms in PMR have not been previously mentioned. This is a first reported case of PMR presented with uveitis, without a complication of GCA. This anterior uveitis might be caused by ischemia of the posterior ciliary arteries and their branches. PMID- 21629813 TI - Galli-galli disease: a rare acantholytic variant of dowling-degos disease. AB - Galli-Galli disease is a rare acantholytic variant of Dowling-Degos disease, with few cases reported in the literature. We describe a case of Galli-Galli disease and review the literature. PMID- 21629814 TI - The effect of gallium nitrate on arresting blood flow from a wound. AB - A novel application of gallium nitrate, hitherto unreported, in reducing bleeding time from an open wound is presented. Experiments performed using simple punctures in the forearm demonstrated a very substantial reduction in bleeding time when a solution of gallium nitrate was applied relative to a control. This outcome was shown to be unaffected by the anticoagulant properties of warfarin. The mechanism for such action of gallium nitrate is unknown and merits further investigation, as do the possibilities for such an application to improve both civilian and defense trauma treatment modalities. PMID- 21629815 TI - Appendicoumbilical fistula: a rare reason for neonatal umbilical mass. AB - The normal umbilicus is a simple structure, but the intrauterine development of the umbilicus is highly complex. Neonatal umbilical mass anomalies usually represent failure of obliteration of the vitelline duct or the allantois which results in persistence of remnants, which can lead to a wide variety of disorders. In this paper, we present a case of an appendicoumbilical fistula in a neonate along with the differential diagnosis and management options. Embryologic explanation of the etiology was discussed with the possible association with different forms of malpositioning and rotation of the gut. PMID- 21629816 TI - Recurrent Incisional Hernia due to Pseudomyxoma Peritonei. AB - Pseudomyxoma peritonei is a rare but challenging neoplastic disease which is characterized with intraperitoneal mucinous-gelatinous fluid accumulation. It rarely presents as a mass mimicking abdominal wall hernias A recurrent incisional hernia due to pseudomyxoma peritonei is presented here. A 60-year-old female patient had been operated on for a left mucinous ovarian cyst 20 cm in diameter in 1998. Mucinous material had disseminated into interloop spaces through the right subdiaphragmatic region. Total abdominal hysterectomy + bilateral salpingooophorectomy and peritoneal toilet had been performed. She was rehospitalized for abdominal distention and a 4 cm defect over the incision and underwent a hernia repair using polypropylene mesh in 2001. Abdominal distention recurred to give a rise to an incisional hernia in 2006. She was reoperated for decompression and repair, but nothing could be done because of sticky adhesions and the incision were simply closed. The patient was referred to our department for operation. A prosthetic hernia repair with 30 * 30 cm polypropylene mesh was performed. The patient was discharged on the postoperative 5th day following an uneventful recovery. However, she died of disseminated disease after 18 months. PMID- 21629817 TI - Granulocytic sarcoma in a nonleukemic patient: place of radiotherapy and systemic therapies. AB - Granulocytic sarcoma is a rare extramedullary tumour, which most often occurs in the course of an acute or chronic leukaemia or myeloproliferative disorders. Rarely it is found before peripheral blood or bone marrow evidence of leukemia is present. We report an unusual case of acute paraplegia at first presentation of a spinal epidural granulocytic sarcoma without any haematological disorder. Therapeutic strategies are discussed in the light of the literature. PMID- 21629818 TI - Cardiac angiosarcoma-associated membranoproliferative glomerulonephropathy. AB - Primary cardiac angiosarcoma is a rare cardiac tumor. The initial clinical course is often asymptomatic, and metastatic disease is present in a majority of affected patients at diagnosis. We present a patient who presented with a hemorrhagic pericardial effusion. No malignant cells were evident on cytological examination. He subsequently developed membranoproliferative glomerulonephritis requiring hemodialysis. Metastatic cardiac angiosarcoma was diagnosed 5 months later. To our knowledge, this is the first reported case of paraneoplastic membranoproliferative glomerulopathy associated with cardiac angiosarcoma. PMID- 21629819 TI - A case of valproate induced hyperammonemic encephalopathy. AB - A 36-years-old man on phenytoin, levetiracetam, and sodium valproate presented with acute confusion. Routine investigations including serum valproate and phenytoin concentration were normal. His serum ammonia concentration was raised. His valproate was held and 2 days later he recovered with concordant normalisation of serum ammonia concentration. Urea acid cycle disorder was ruled out, and a diagnosis of valproate induced hyperammonemic encephalopathy (VHE) was made. Asymptomatic hyperammonemia occurs in 15-50% of valproate-treated patients, and while the true incidence of VHE is not known, it is a recognized complication of sodium valproate treatment. VHE typically presents acutely with impaired consciousness, lethargy, and vomiting. Valproate concentrations may be in the therapeutic range, and liver function tests are typically "normal." Treatment for VHE consists of ceasing valproate and providing supportive care. Some have advocated carnitine replacement. PMID- 21629820 TI - Bilateral Peritonsillar Abscesses: A Case Presentation and Review of the Current Literature with regard to the Controversies in Diagnosis and Treatment. AB - Although unilateral peritonsillar abscess is a common complication of acute bacterial tonsillitis, bilateral peritonsillar abscesses are quite rare. The incidence of unsuspected contralateral peritonsillar abscess identified at tonsillectomy has been reported to be between 1.9% and 24%, while the overall incidence of bilateral peritonsillar abscess is reported to reach 4.9%. Diagnosis can be based on clinical criteria or imaging techniques. As far as the treatment is concerned, it is generally accepted that the basic strategy consists of systemic antibiotics and drainage of the pus. We report the case of a 19-year-old girl, treated in the emergency room with a bilateral diagnostic needle aspiration followed by bilateral incision and drainage along with intravenous clindamycin plus anti-inflammatory agents and hydration. Following treatment, the patient progressively experienced a marked alleviation of her odynophagia. She was discharged 48 hours later on a 10-day course of clindamycin. PMID- 21629822 TI - Innate neural assemblies for lego memory. PMID- 21629821 TI - Does reconsolidation occur in humans? AB - Evidence for reconsolidation in non-human animals has accumulated rapidly in the last decade, providing compelling' demonstration for this phenomenon across species and memory paradigms. In vast contrast, scant evidence exists for human reconsolidation to date. A major reason for this discrepancy is the invasive nature of current techniques used to investigate reconsolidation, which are difficult to apply in humans. Pharmacological blockade of reconsolidation, for example, has been typically used in animals as a proof of concept. However, most compounds used in these studies are toxic for humans, and those compounds that are safe target related, but not direct mechanisms of reconsolidation. Thus, although human reconsolidation has been hypothesized, there is limited evidence it actually exists. The best evidence for human reconsolidation emerges from non invasive techniques that "update" memory during reconsolidation rather than block it, a technique only rarely used in animal research. Here we discuss the current state of human reconsolidation and the challenges ahead. We review findings on reconsolidation of emotional associative, episodic, and procedural memories, using invasive and non-invasive techniques. We discuss the possible interpretation of these results, attempt to reconcile some inconsistencies, and suggest a conceptual framework for future research. PMID- 21629823 TI - Transcription Mapping of Embryonic Rat Brain Reveals EGR-1 Induction in SOX2 Neural Progenitor Cells. AB - Neuronal expression of the early growth response-1 (EGR-1; NGFI-A/Zif268) transcription factor has been extensively studied in the adult mammalian brain and linked to aspects of mature physiological/behavioral function. In contrast, this factor has not been studied in detail in the embryonic brain. Here, we used a fluorescent protein-encoding Egr-1 transgene to map the cellular distribution of Egr-1 transcription in embryonic rat brain. We identified a novel, widely distributed population of GFP(+) cells, characterized as a precursor/stem cell phenotype by co-localization with SOX2/nestin/vimentin/S-100beta and exclusion from other known cellular markers including DCX/BLBP/TBR2/NURR1. At both E18 and E20, these cells were located across the developing brain but concentrated in the subplate and intermediate zones. The transgene was also highly expressed in developing (NeuN(+)) striatal neurons. The authentic expression pattern that we observed for the rEgr-1 transgene sequence indicates that restriction to neuronal/precursor cells is largely driven by proximal 5(') sequence. Deletion of conserved Egr-1 silencer (neuron restrictive silencer factor) elements did not markedly alter transcriptional activity in transfected cells; this is consistent with a dominant role for positive factors in the control of cell-specific Egr-1 expression. Induction of Egr-1 in a population of SOX2(+) cells indicates a co incidence of extrinsic (EGR-1) and cell-intrinsic (SOX2) cellular signals that may form a novel level of progenitor cell regulation. The wide distribution of EGR-1 signaling in SOX2(+) cells suggests an organizational role during late embryonic brain development. PMID- 21629824 TI - Habituation without NMDA Receptor-Dependent Desensitization of Hering-Breuer Apnea Reflex in a Mecp2 Mutant Mouse Model of Rett Syndrome. AB - Non-associative learning is a basic neuroadaptive behavior exhibited in almost all animal species and sensory modalities but its functions and mechanisms in the mammalian brain are poorly understood. Previous studies have identified two distinct forms of non-associative learning in the classic Hering-Breuer inflation reflex (HBIR) induced apnea in rats: NMDA receptor (NMDAR)-independent habituation in a primary vagal pathway and NMDAR-dependent desensitization in a secondary pontine pathway. Here, we show that abnormal non-associative learning of the HBIR may underlie the endophenotypic tachypnea in an animal model of Rett syndrome (RTT), an autism-spectrum disorder caused by mutations in the X-linked gene encoding methyl-CpG-binding protein 2 (MECP2). Mecp2(+/-) symptomatic mice on a mixed-strain background demonstrated significantly increased resting respiratory frequency with shortened expiration and normal inspiratory duration compared with asymptomatic mutants and wild-type controls, a phenotype that is characteristic of girls with RTT. Low-intensity electrical stimulation of the vagus nerve elicited fictive HBIR with time-dependent habituation in both Mecp2(+/-) and wild-type mice. However, time-dependent desensitization of the HBIR was evidenced only in wild-type controls and asymptomatic mutant mice but was absent or suppressed in Mecp2(+/-) symptomatic mice or in wild-type mice after blockade of NMDAR with dizocilpine. Remarkably, ~50% of the Mecp2(+/-) mice developed these X-linked phenotypes despite somatic mosaicism. Such RTT-like respiratory endophenotypes in mixed-strain Mecp2(+/-) mice differed from those previously reported in Mecp2(-/y) mice on pure C57BL/6J background. These findings provide the first evidence indicating that impaired NMDAR-dependent desensitization of the HBIR may contribute to the endophenotypic tachypnea in RTT. PMID- 21629825 TI - Signaling in migrating neurons: from molecules to networks. AB - During prenatal and postnatal development of the mammalian brain, new neurons are generated by precursor cells that are located in the germinal zones. Subsequently newborn neurons migrate to their destined location in the brain. On the migrational route immature neurons interact via a series of recognition molecules with a plethora of extracellular cues. Stimuli that are conveyed by extracellular cues are translated into complex intracellular signaling networks that eventually enable neuronal migration. In this Focused Review we discuss signaling networks underlying neuronal migration emphasizing molecules and pathways that appear to be neuron-specific. PMID- 21629826 TI - A bayesian foundation for individual learning under uncertainty. AB - Computational learning models are critical for understanding mechanisms of adaptive behavior. However, the two major current frameworks, reinforcement learning (RL) and Bayesian learning, both have certain limitations. For example, many Bayesian models are agnostic of inter-individual variability and involve complicated integrals, making online learning difficult. Here, we introduce a generic hierarchical Bayesian framework for individual learning under multiple forms of uncertainty (e.g., environmental volatility and perceptual uncertainty). The model assumes Gaussian random walks of states at all but the first level, with the step size determined by the next highest level. The coupling between levels is controlled by parameters that shape the influence of uncertainty on learning in a subject-specific fashion. Using variational Bayes under a mean field approximation and a novel approximation to the posterior energy function, we derive trial-by-trial update equations which (i) are analytical and extremely efficient, enabling real-time learning, (ii) have a natural interpretation in terms of RL, and (iii) contain parameters representing processes which play a key role in current theories of learning, e.g., precision-weighting of prediction error. These parameters allow for the expression of individual differences in learning and may relate to specific neuromodulatory mechanisms in the brain. Our model is very general: it can deal with both discrete and continuous states and equally accounts for deterministic and probabilistic relations between environmental events and perceptual states (i.e., situations with and without perceptual uncertainty). These properties are illustrated by simulations and analyses of empirical time series. Overall, our framework provides a novel foundation for understanding normal and pathological learning that contextualizes RL within a generic Bayesian scheme and thus connects it to principles of optimality from probability theory. PMID- 21629827 TI - Conservative decisions guided by the anterior cingulate cortex. PMID- 21629828 TI - Three-dimensional reconstruction of serial mouse brain sections: solution for flattening high-resolution large-scale mosaics. AB - Recent advances in high-throughput technology facilitate massive data collection and sharing, enabling neuroscientists to explore the brain across a large range of spatial scales. One such form of high-throughput data collection is the construction of large-scale mosaic volumes using light microscopy (Chow et al., 2006; Price et al., 2006). With this technology, researchers can collect and analyze high-resolution digitized volumes of whole brain sections down to 0.2 MUm. However, until recently, scientists lacked the tools to easily handle these large high-resolution datasets. Furthermore, artifacts resulting from specimen preparation limited volume reconstruction using this technique to only a single tissue section. In this paper, we carefully describe the steps we used to digitally reconstruct a series of consecutive mouse brain sections labeled with three stains, a stain for blood vessels (DiI), a nuclear stain (TO-PRO-3), and a myelin stain (FluoroMyelin). These stains label important neuroanatomical landmarks that are used for stacking the serial sections during reconstruction. In addition, we show that the use of two software applications, ir-Tweak and Mogrifier, in conjunction with a volume flattening procedure enable scientists to adeptly work with digitized volumes despite tears and thickness variations within tissue sections. These applications make processing large-scale brain mosaics more efficient. When used in combination with new database resources, these brain maps should allow researchers to extend the lifetime of their specimens indefinitely by preserving them in digital form, making them available for future analyses as our knowledge in the field of neuroscience continues to expand. PMID- 21629829 TI - Meiofauna in the Gollum Channels and the Whittard Canyon, Celtic Margin--how local environmental conditions shape nematode structure and function. AB - The Gollum Channels and Whittard Canyon (NE Atlantic) are two areas that receive high input of organic matter and phytodetritus from euphotic layers, but they are typified by different trophic and hydrodynamic conditions. Sediment biogeochemistry was analysed in conjunction with structure and diversity of the nematode community and differences were tested between study areas, water depths (700 m vs 1000 m), stations, and sediment layers. The Gollum Channels and Whittard Canyon harboured high meiofauna abundances (1054-1426 ind. 10 cm(-2)) and high nematode diversity (total of 181 genera). Next to enhanced meiofauna abundance and nematode biomass, there were signs of high levels of organic matter deposition leading to reduced sedimentary conditions, which in turn structured the nematode community. Striking in this respect was the presence of large numbers of 'chemosynthetic' Astomonema nematodes (Astomonema southwardorum, Order Monhysterida, Family Siphonolaimidae). This genus lacks a mouth, buccal cavity and pharynx and possesses a rudimentary gut containing internal, symbiotic prokaryotes which have been recognised as sulphur-oxidising bacteria. Dominance of Astomonema may indicate the presence of reduced environments in the study areas, which is partially confirmed by the local biogeochemical environment. The nematode communities were mostly affected by sediment layer differences and concomitant trophic conditions rather than other spatial gradients related to study area, water depth or station differences, pointing to small-scale heterogeneity as the main source of variation in nematode structure and function. Furthermore, the positive relation between nematode standing stocks, and quantity and quality of the organic matter was stronger when hydrodynamic disturbance was greater. Analogically, this study also suggests that structural diversity can be positively correlated with trophic conditions and that this relation is tighter when hydrodynamic disturbance is greater. PMID- 21629830 TI - Capecitabine in the management of colorectal cancer. AB - 5-Fluorouracil has been a mainstay in the treatment of colorectal cancer for nearly five decades; however, the use of oral formulations of the medication has been gaining increasing traction since capecitabine was approved for use in adjuvant settings by the US Food and Drug Administration in 2005. The use of capecitabine has since spread to a number of off-label indications, including the treatment of advanced or metastatic colorectal cancer and the neoadjuvant treatment of rectal cancer. In light of increasing utilization, it is critical that clinicians have a firm understanding of the literature supporting capecitabine across various settings as well as the attributes of the drug, such as its dosing recommendations, side-effect profile, and use in the elderly. The purpose of this review is to synthesize the literature in a fashion that can be used to help guide decisions. In a setting of increasing focus on cost, the pharmacoeconomic literature is also briefly reviewed. PMID- 21629831 TI - Chemoprevention of prostate cancer with nutrients and supplements. AB - As the adult population is increasing, prostate cancer (PCa) will become a considerable health problem in the next millennium. This has raised public interest in potential chemoprevention of this disease. As PCa is extremely common and generally slow to progress it is regarded as an ideal candidate for chemoprevention. At present, the 5 alpha-reductase inhibitors finasteride and dutasteride have been identified as preventive agents. This review describes whether selenium, alpha-tocopherol, isoflavones, lycopene green tea polyphenols, calcium, and resveratrol may be useful for decreasing the risk of PCa in men. Although encouraging results are present, some studies show negative results. Differences in study design, sample size, dose administered, and/or concentrations achieved in the body may be the reason for these inconsistencies. Today, chemopreventive agents may be appropriate for high-risk patients like those with high-grade prostatic intraepithelial neoplasia and other high-risk groups such as patients with elevated prostate specific antigen (PSA) and negative biopsy, rapid PSA velocity, and with a family history of PCa. Although larger randomized controlled studies are needed and epidemiologic evidence should be placed in a clinical context, physicians must be aware of these preventive opportunities in PCa care. Combinations of chemopreventive agents should be carefully investigated because mechanisms of action may be additive or synergistic. PMID- 21629832 TI - Improving evaluation of the distribution and density of immunostained cells in breast cancer using computerized video image analysis. AB - Quantitation of cell density in tissues has proven problematic over the years. The manual microscopic methodology, where an investigator visually samples multiple areas within slides of tissue sections, has long remained the basic 'standard' for many studies and for routine histopathologic reporting. Nevertheless, novel techniques that may provide a more standardized approach to quantitation of cells in tissue sections have been made possible by computerized video image analysis methods over recent years. The present study describes a novel, computer-assisted video image analysis method of quantitating immunostained cells within tissue sections, providing continuous graphical data. This technique enables the measurement of both distribution and density of cells within tissue sections. Specifically, the study considered immunoperoxidase stained tumor infiltrating lymphocytes within breast tumor specimens, using the number of immunostained pixels within tissue sections to determine cellular density and number. Comparison was made between standard manual graded quantitation methods and video image analysis, using the same tissue sections. The study demonstrates that video image techniques and computer analysis can provide continuous data on cell density and number in immunostained tissue sections, which compares favorably with standard visual quantitation methods, and may offer an alternative. PMID- 21629833 TI - Bioimpedance and chronoamperometry as an adjunct to prostate-specific antigen screening for prostate cancer. AB - BACKGROUND: Bioimpedance is an electrical property of living tissue that has been shown to be a safe technique when used in a number of biomedical applications. The aim of this research was to assess the utility of bioimpedance measurement as a rapid, cost-effective, and noninvasive adjunct to digital rectal examination and PSA in differentiating tumor from normal prostatic tissue. METHODS: Three hundred men were examined for signs and symptoms of prostate disorders. 147 patients with a digital rectal examination indicating a positive result underwent a prostate-specific antigen (PSA) test. A biopsy was advised for 103 of the men, of whom 50 completed the study. Before undergoing biopsy, an examination with the EIS (electro interstitial scan) system using bioimpedance and chronoamperometry was performed. In reference to the biopsy results (negative or positive), a statistical analysis of the EIS data and PSA was conducted using receiver operating characteristic curves to determine the specificity and sensitivity of each test. RESULTS: The PSA test had a sensitivity of 73.9% and specificity of 51.9% using a cutoff value >4 and a sensitivity of 52.2% and specificity of 81.5% using a cutoff value >=5.7 and P = 0.03. The delta of the electrical conductivity (DE) of the left foot-right foot pathway had a sensitivity of 62.5% and specificity of 85.2%, with a cutoff value <=-5 and P = 0.0001. Algorithms comprising the delta of electrical conductivity and PSA showed a sensitivity of 91.5% and a specificity of 59.3%, with a cutoff value <=-10.52 and P = 0.0003. CONCLUSION: The EIS system had a very good specificity of 85.2%. However, the sensitivity of 62.5% would be a problem. Using a PSA reference >4.1 ng/mL, the adjunctive use of bioimpedance and chronoamperometry provided by EIS technology could raise the sensitivity from 73.9% to 91.5% and the specificity from 51.9% to 59.3% in prostate cancer screening. PMID- 21629834 TI - Widespread occurrence of secondary lipid biosynthesis potential in microbial lineages. AB - Bacterial production of long-chain omega-3 polyunsaturated fatty acids (PUFAs), such as eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n 3), is constrained to a narrow subset of marine gamma-proteobacteria. The genes responsible for de novo bacterial PUFA biosynthesis, designated pfaEABCD, encode large, multi-domain protein complexes akin to type I iterative fatty acid and polyketide synthases, herein referred to as "Pfa synthases". In addition to the archetypal Pfa synthase gene products from marine bacteria, we have identified homologous type I FAS/PKS gene clusters in diverse microbial lineages spanning 45 genera representing 10 phyla, presumed to be involved in long-chain fatty acid biosynthesis. In total, 20 distinct types of gene clusters were identified. Collectively, we propose the designation of "secondary lipids" to describe these biosynthetic pathways and products, a proposition consistent with the "secondary metabolite" vernacular. Phylogenomic analysis reveals a high degree of functional conservation within distinct biosynthetic pathways. Incongruence between secondary lipid synthase functional clades and taxonomic group membership combined with the lack of orthologous gene clusters in closely related strains suggests horizontal gene transfer has contributed to the dissemination of specialized lipid biosynthetic activities across disparate microbial lineages. PMID- 21629835 TI - How Should Addiction-Related Research at the National Institutes of Health be Reorganized? PMID- 21629836 TI - Maternal genetic mutations as gestational and early life influences in producing psychiatric disease-like phenotypes in mice. AB - Risk factors for psychiatric disorders have traditionally been classified as genetic or environmental. Risk (candidate) genes, although typically possessing small effects, represent a clear starting point to elucidate downstream cellular/molecular pathways of disease. Environmental effects, especially during development, can also lead to altered behavior and increased risk for disease. An important environmental factor is the mother, demonstrated by the negative effects elicited by maternal gestational stress and altered maternal care. These maternal effects can also have a genetic basis (e.g., maternal genetic variability and mutations). The focus of this review is "maternal genotype effects" that influence the emotional development of the offspring resulting in life-long psychiatric disease-like phenotypes. We have recently found that genetic inactivation of the serotonin 1A receptor (5-HT1AR) and the fmr1 gene (encoding the fragile X mental retardation protein) in mouse dams results in psychiatric disease-like phenotypes in their genetically unaffected offspring. 5 HT1AR deficiency in dams results in anxiety and increased stress responsiveness in their offspring. Offspring of 5-HT1AR deficient dams display altered development of the hippocampus, which could be linked to their anxiety-like phenotype. Maternal inactivation of fmr1, like its inactivation in the offspring, results in a hyperactivity-like condition and is associated with receptor alterations in the striatum. These data indicate a high sensitivity of the offspring to maternal mutations and suggest that maternal genotype effects can increase the impact of genetic risk factors in a population by increasing the risk of the genetically normal offspring as well as by enhancing the effects of offspring mutations. PMID- 21629837 TI - Addiction, adolescence, and innate immune gene induction. AB - Repeated drug use/abuse amplifies psychopathology, progressively reducing frontal lobe behavioral control, and cognitive flexibility while simultaneously increasing limbic temporal lobe negative emotionality. The period of adolescence is a neurodevelopmental stage characterized by poor behavioral control as well as strong limbic reward and thrill seeking. Repeated drug abuse and/or stress during this stage increase the risk of addiction and elevate activator innate immune signaling in the brain. Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kappaB) is a key glial transcription factor that regulates proinflammatory chemokines, cytokines, oxidases, proteases, and other innate immune genes. Induction of innate brain immune gene expression (e.g., NF-kappaB) facilitates negative affect, depression-like behaviors, and inhibits hippocampal neurogenesis. In addition, innate immune gene induction alters cortical neurotransmission consistent with loss of behavioral control. Studies with anti oxidant, anti-inflammatory, and anti-depressant drugs as well as opiate antagonists link persistent innate immune gene expression to key behavioral components of addiction, e.g., negative affect-anxiety and loss of frontal cortical behavioral control. This review suggests that persistent and progressive changes in innate immune gene expression contribute to the development of addiction. Innate immune genes may represent a novel new target for addiction therapy. PMID- 21629838 TI - Increasing Opportunity through Interdisciplinary Research: Climbing Down and Shattering a Tower of Babel. PMID- 21629839 TI - Litter gender composition and sex affect maternal behavior and DNA methylation levels of the oprm1 gene in rat offspring. AB - The mu-opioid receptor is encoded by the Oprm1 gene and contributes to mother infant behaviors. Rodent dams lick male pups more than female pups in the anogenital region. This behavior is linked to stress responsivity in the offspring that may be mediated by epigenetic changes. We hypothesized that maternal behavior may affect DNA methylation levels of the Oprm1 gene and show sex differences. To further explore sex differences in mother-pup behaviors and DNA methylation levels, we altered the litter gender composition (LGC) of rats. Litters were culled to eight all male, all female, or four male/four female pups on postnatal (PN) day 1. On PN4, 7, and 10, a dam was placed in a test cage with a pup for a 10-min period. Latency to pup contact was determined as were times spent licking the anogenital and other body regions of the pup. Frequencies of other behaviors were tabulated. On PN35, samples from various brain regions were obtained. DNA methylation at specific CpG sites in the Oprm1 promoter region were measured by direct sequencing of bisulfite-treated DNA. LGC and sex interacted with day for latency to pup contact. Latencies were longest on PN4 for single-sex males and on PN10 for single-sex females. Dams licked male pups more than female pups in both the anogenital and other body areas. Sex differences were seen in other behaviors. LGC altered DNA methylation at specific CpG's of Oprm1 in hippocampus with higher levels in single-sex rats. In nucleus accumbens, single sex males showed hypermethylation levels, a trend seen in caudate-putamen. Results confirm and extend sex differences in maternal care with modest LGC effects. That both LGC and sex have enduring effects on DNA methylation of the Oprm1 gene in brain regions associated with addiction, stress regulation, motivation, and cognition may suggest one factor that contributes to gender differences in these behaviors. PMID- 21629840 TI - Aberrant NF-kappaB expression in autism spectrum condition: a mechanism for neuroinflammation. AB - Autism spectrum condition (ASC) is recognized as having an inflammatory component. Post-mortem brain samples from patients with ASC display neuroglial activation and inflammatory markers in cerebrospinal fluid, although little is known about the underlying molecular mechanisms. Nuclear factor kappa-light-chain enhancer of activated B cells (NF-kappaB) is a protein found in almost all cell types and mediates regulation of immune response by inducing the expression of inflammatory cytokines and chemokines, establishing a feedback mechanism that can produce chronic or excessive inflammation. This article describes immunodetection and immunofluorescence measurements of NF-kappaB in human post-mortem samples of orbitofrontal cortex tissue donated to two independent centers: London Brain Bank, Kings College London, UK (ASC: n = 3, controls: n = 4) and Autism Tissue Program, Harvard Brain Bank, USA (ASC: n = 6, controls: n = 5). The hypothesis was that concentrations of NF-kappaB would be elevated, especially in activated microglia in ASC, and pH would be concomitantly reduced (i.e., acidification). Neurons, astrocytes, and microglia all demonstrated increased extranuclear and nuclear translocated NF-kappaB p65 expression in brain tissue from ASC donors relative to samples from matched controls. These between-groups differences were increased in astrocytes and microglia relative to neurons, but particularly pronounced for highly mature microglia. Measurement of pH in homogenized samples demonstrated a 0.98-unit difference in means and a strong (F = 98.3; p = 0.00018) linear relationship to the expression of nuclear translocated NF-kappaB in mature microglia. Acridine orange staining localized pH reductions to lysosomal compartments. In summary, NF-kappaB is aberrantly expressed in orbitofrontal cortex in patients with ASC, as part of a putative molecular cascade leading to inflammation, especially of resident immune cells in brain regions associated with the behavioral and clinical symptoms of ASC. PMID- 21629841 TI - Are behavioral effects of early experience mediated by oxytocin? AB - Early experiences can alter adaptive emotional responses necessary for social behavior as well as physiological reactivity in the face of challenge. In the highly social prairie vole (Microtus ochrogaster), manipulations in early life or hormonal treatments specifically targeted at the neuropeptides oxytocin (OT) and arginine vasopressin (AVP), have long-lasting, often sexually dimorphic, consequences for social behavior. Here we examine the hypothesis that behavioral changes associated with differential early experience, in this case handling the family during the first week of life, may be mediated by changes in OT or AVP or their brain receptors. Four early treatment groups were used, differing only in the amount of manipulation received during the first week of life. MAN1 animals were handled once on post-natal day 1; MAN1 treatment produces a pattern of behavior usually considered typical of this species, against which other groups were compared. MAN1-7 animals were handled once a day for post-natal days 1-7, MAN 7 animals were handled once on post-natal day 7, and MAN0 animals received no handling during the first week of life. When tested following weaning, males in groups that had received manipulation during the first few days of life (MAN1 and MAN1-7) displayed higher alloparenting than other groups. Neuroendocrine measures, including OT receptor binding and OT and AVP immunoreactivity, varied by early treatment. In brain areas including the nucleus accumbens, bed nucleus of stria terminalis and lateral septum, MAN0 females showed increased OT receptor binding. MAN1 animals also displayed higher numbers of immunoreactive OT cell bodies in the supraoptic nucleus. Taken together these findings support the broader hypothesis that experiences in the first few days of life, mediated in part by sexually dimorphic changes in neuropeptides, especially in the receptor for OT, may have adaptive consequences for sociality and emotion regulation. PMID- 21629842 TI - Incorporating genetics into your studies: a guide for social scientists. AB - There has been a surge of interest in recent years in incorporating genetic components into on-going longitudinal, developmental studies and related psychological studies. While this represents an exciting new direction in developmental science, much of the research on genetic topics in developmental science does not reflect the most current practice in genetics. This is likely due, in part, to the rapidly changing landscape of the field of genetics, and the difficulty this presents for developmental scientists who are trying to learn this new area. In this review, we present an overview of the paradigm shifts that have occurred in genetics and we introduce the reader to basic genetic methodologies. We present our view of the current stage of research ongoing at the intersection of genetics and social science, and we provide recommendations for how we could do better. We also address a number of issues that social scientists face as they integrate genetics into their projects, including choice of a study design (candidate gene versus genome-wide association versus sequencing), different methods of DNA collection, and special considerations involved in the analysis of genotypic data. Through this review, we hope to equip social scientists with a deeper understanding of the many considerations that go into genetics research, in an effort to foster more meaningful cross-disciplinary initiatives. PMID- 21629843 TI - Functional genomic and proteomic analysis reveals disruption of myelin-related genes and translation in a mouse model of early life neglect. AB - Early life neglect is an important public health problem which can lead to lasting psychological dysfunction. Good animal models are necessary to understand the mechanisms responsible for the behavioral and anatomical pathology that results. We recently described a novel model of early life neglect, maternal separation with early weaning (MSEW), that produces behavioral changes in the mouse that persist into adulthood. To begin to understand the mechanism by which MSEW leads to these changes we applied cDNA microarray, next-generation RNA sequencing (RNA-seq), label-free proteomics, multiple reaction monitoring (MRM) proteomics, and methylation analysis to tissue samples obtained from medial prefrontal cortex to determine the molecular changes induced by MSEW that persist into adulthood. The results show that MSEW leads to dysregulation of markers of mature oligodendrocytes and genes involved in protein translation and other categories, an apparent downward biasing of translation, and methylation changes in the promoter regions of selected dysregulated genes. These findings are likely to prove useful in understanding the mechanism by which early life neglect affects brain structure, cognition, and behavior. PMID- 21629844 TI - Mesolimbic dopamine transients in motivated behaviors: focus on maternal behavior. AB - Phasic activity of the mesolimbic dopamine pathway - burst-firing of dopamine neurons and the resulting dopamine release events at striatal targets - have been associated with a variety of motivational events, such as novelty, salient stimuli, social interaction, and reward prediction. Over the past decade, advances in electrochemical techniques have allowed measurement of naturally occurring dopamine release events, or dopamine transients, in awake animals during ongoing behavior. Thus, a growing body of studies has revealed dynamic dopamine input to ventral striatum during motivated behavior in a variety of experimental paradigms. We propose that dopamine transients may be important neural signals in pup-directed aspects of maternal behavior, as preliminary data suggest that dopamine transients in dams are associated with pup cues. Measurements of dopamine transients may be useful to investigate not only typical maternal behavior but also maternal inattention induced by drug exposure or stress. PMID- 21629845 TI - Evolution of Substance use, Neurological and Psychiatric Symptoms in Schizophrenia and Substance use Disorder Patients: A 12-Week, Pilot, Case-Control Trial with Quetiapine. AB - Neurological and psychiatric symptoms are consequences of substance abuse in schizophrenia and non-schizophrenia patients. The present case-control study examined changes in substance abuse/dependence, and neurological and psychiatric symptoms in substance abusers with [dual diagnosis (DD) group, n = 26] and without schizophrenia [substance use disorder (SUD) group, n = 24] and in non abusing schizophrenia patients (SCZ group, n = 23) undergoing 12-week treatment with the atypical antipsychotic, quetiapine. Neurological and psychiatric symptoms were evaluated with the Positive and Negative Syndrome Scale, the Calgary Depression Scale for Schizophrenia, the Extrapyramidal Symptoms Rating Scale, and the Barnes Akathisia Rating Scale. At endpoint, DD and SCZ patients were receiving significantly higher doses of quetiapine (mean = 554 and 478 mg/day, respectively), relative to SUD patients (mean = 150 mg/day). We found that SUD patients showed greater improvement in weekly dollars spent on alcohol and drugs and SUD severity, compared to DD patients. At endpoint, there was no significant difference in dollars spent, but DD patients still had a higher mean SUD severity. Interestingly, DD patients had significantly higher parkinsonism and depression than SCZ patients at baseline and endpoint. On the other hand, we found that SUD patients had significantly more akathisia at baseline, improved more than SCZ patients, and this was related to cannabis abuse/dependence. Finally, SUD patients improved more in Positive and Negative Syndrome Scale positive scores than DD and SCZ patients. Taken together, our results provide evidence for increased vulnerability to the adverse effects of alcohol and drugs in schizophrenia patients. They also suggest that substance abuse/withdrawal may mimic some symptoms of schizophrenia. Future studies will need to determine the role quetiapine played in these improvements. PMID- 21629846 TI - Dramatic response of nail psoriasis to infliximab. AB - Nail psoriasis, affecting up to 50% of psoriatic patients, is an important cause of serious psychological and physical distress. Traditional treatments for nail psoriasis, which include topical or intralesional corticosteroids, topical vitamin D analogues, photochemotherapy, oral retinoids, methotrexate, and cyclosporin, can be time-consuming, painful, or limited by significant toxicities. Biological agents may have the potential to revolutionize the management of patients with disabling nail psoriasis. We present another case of disabling nail psoriasis that responded dramatically to infliximab. PMID- 21629847 TI - Comparison of the in vivo distribution of four different annexin a5 adducts in rhesus monkeys. AB - Annexin A5 has been used for the detection of apoptotic cells, due to its ability to bind to phosphatidylserine (PS). Four different labeled Annexin A5 adducts were evaluated in rhesus monkey, with radiolabeling achieved via 1,4,7,10 tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA). Of these adducts differing conjugation methods were employed which resulted in nonspecific radiolabeling (AxA5-I), or site-specific radiolabeling (AxA5-II). A nonbinding variant of Annexin A5 was also evaluated (AxA5-II(NBV)), conjugation here was site specific. The fourth adduct examined had both specific and nonspecific conjugation techniques employed (AxA5-II(mDOTA)). Blood clearance for each adduct was comparable, while appreciable uptake was observed in kidney, liver, and spleen. Significant differences in uptake of AxA5-I and AxA5-II were observed, as well as between AxA5-II and AxA5-II(NBV). No difference between AxA5-II and AxA5 II(mDOTA) was observed, suggesting that conjugating DOTA nonspecifically did not affect the in vivo biodistribution of Annexin A5. PMID- 21629848 TI - Phantom limb pain: low frequency repetitive transcranial magnetic stimulation in unaffected hemisphere. AB - Phantom limb pain is very common after limb amputation and is often difficult to treat. The motor cortex stimulation is a valid treatment for deafferentation pain that does not respond to conventional pain treatment, with relief for 50% to 70% of patients. This treatment is invasive as it uses implanted epidural electrodes. Cortical stimulation can be performed noninvasively by repetitive transcranial magnetic stimulation (rTMS). The stimulation of the hemisphere that isn't involved in phantom limb (unaffected hemisphere), remains unexplored. We report a case of phantom limb pain treated with 1 Hz rTMS stimulation over motor cortex in unaffected hemisphere. This stimulation produces a relevant clinical improvement of phantom limb pain; however, further studies are necessary to determine the efficacy of the method and the stimulation parameters. PMID- 21629849 TI - In vitro and ex vivo evaluation of smart infra-red fluorescent caspase-3 probes for molecular imaging of cardiovascular apoptosis. AB - Purpose. The aim of this paper is to develop new optical bioprobes for the imaging of apoptosis. Procedure. We developed quenched near-infrared probes which become fluorescent upon cleavage by caspase-3, the key regulatory enzyme of apoptosis. Results. Probes were shown to be selectively cleaved by recombinant caspase-3. Apoptosis of cultured endothelial cells was associated with an increased fluorescent signal for the cleaved probes, which colocalized with caspase-3 and was reduced by the addition of a caspase-3 inhibitor. Flow cytometry demonstrated a similar profile between the cleaved probes and annexin V. Ex vivo experiments showed that sections of hearts obtained from mice treated with the proapoptotic drug doxorubicin displayed an increase in the fluorescent signal for the cleaved probes, which was reduced by a caspase-3 inhibitor. Conclusion. We demonstrated the capacity of these novel probes to detect apoptosis by optical imaging in vitro and ex vivo. PMID- 21629850 TI - Systemic Bartonella henselae Infection in Immunocompetent Adult Presenting as Fever of Unknown Origin. AB - Systemic clinical presentations of infection caused by Bartonella henselae are rare in immunocompetent adults. We report four cases with hepatic and/or splenic involvement, presenting as fever of unknown origin. We discuss diagnosis and treatment of this infection. Bartonella henselae serology allows an easy diagnosis of hepatosplenic involvement in cat scratch disease, a clinical picture that appears to be underrecognized. PMID- 21629851 TI - Renal function but not asymmetric dimethylarginine is independently associated with retinopathy in type 2 diabetes. AB - Background. Asymmetric dimethylarginine (ADMA) is associated with macrovascular disease and possibly with microangiopathy in type 2 diabetes (T2DM). We tested the hypothesis that ADMA is related to diabetic retinopathy (DR) independently of macrovascular disease. Methods. This cross-sectional study included 127 T2DM patients selected to achieve equal distributions of patients with and without macrovascular disease in the groups with and without DR. Results. Patients with DR had increased ADMA, longer diabetes duration, and reduced glomerular filtration rate (GFR). ADMA correlated with GFR (rho = -0.35; P < .001), diabetes duration (rho = 0.19; P = .048), and age (rho = 0.19; P = .033). Logistic regression analysis revealed an association of ADMA with DR. After adjustment for macrovascular disease, this association remained significant (OR 1.48; 95% CI: 1.02-2.15; P = .039). Inclusion of GFR and T2DM duration into the model abolished this significant relationship. GFR remained the only independent predictor for DR. A 10 mL/min/1.73 m(2) GFR decrease was associated with DR in a multivariate model (OR 1.30; 95% CI: 1.08-1.56; P = .006). Conclusions. These findings indicate an association between ADMA and DR in T2DM independent of macrovascular disease. This relationship is modified by GFR, the only parameter significantly related to DR in multivariate analysis. PMID- 21629852 TI - A case of unresectable rectal necrosis. AB - Introduction. Necrosis of the rectum is an uncommon finding due to abundant collateral vasculature. Its management remains challenging, without clear consensus in the literature. Case Report. We describe a case of a 53-year-old woman with multiple medical comorbidities that presented in septic shock and hematochezia. Colonoscopy revealed ischemic colitis. Conservative management was instituted. At two weeks, she presented evidence of peritonitis. Exploratory laparotomy revealed extensive necrosis of the left colon and rectum. Due to dense inflammation, resection was deemed unsafe. Therefore, a transverse ostomy with mucosal fistula was preformed. Multiple drains were left in place. The patient healed uneventfully. Conclusion. This case illustrates that, if extensive dissection of the distal colon and rectum is unsafe due to the patient's critical condition or technical feasibility, then a diverting ostomy of the proximal viable bowel along with a mucus fistula and good drainage of the abdomen represents an acceptable alternative. PMID- 21629853 TI - Odontogenic fibromyxoma of the maxilla: a case report and review of the literature. AB - Fibromyxoma represents a rare benign neoplasm that mostly affects the posterior region of the mandible. Here, we report the case of a 46-year-old male with a swelling of the right maxilla. After proper diagnosis, he was treated with enucleation and curettage of the tumor. The defect was filled with a pedicled buccal fat pad flap. The mesenchymal origin from the dental follicle of the fibromyxoma is the most plausible explanation. Radiological examination with MRI, CT, and conventional radiography contributes to the differential diagnosis from other benign tumors, such as the ameloblastoma. Its management is surgical and comprises enucleation and curettage or en bloc resection. Patients must be monitored for at least two years postoperatively in order to diagnose possible recurrence. According to the literature, the maxilla is a rare location of a fibromyxoma and, to our knowledge, our case is the 30th presented case of a fibromyxoma of the maxilla. PMID- 21629856 TI - The Neurological Ecology of Fear: Insights Neuroscientists and Ecologists Have to Offer one Another. AB - That the fear and stress of life-threatening experiences can leave an indelible trace on the brain is most clearly exemplified by post-traumatic stress disorder (PTSD). Many researchers studying the animal model of PTSD have adopted utilizing exposure to a predator as a life-threatening psychological stressor, to emulate the experience in humans, and the resulting body of literature has demonstrated numerous long-lasting neurological effects paralleling those in PTSD patients. Even though much more extreme, predator-induced fear and stress in animals in the wild was, until the 1990s, not thought to have any lasting effects, whereas recent experiments have demonstrated that the effects on free-living animals are sufficiently long-lasting to even affect reproduction, though the lasting neurological effects remain unexplored. We suggest neuroscientists and ecologists both have much to gain from collaborating in studying the neurological effects of predator-induced fear and stress in animals in the wild. We outline the approaches taken in the lab that appear most readily translatable to the field, and detail the advantages that studying animals in the wild can offer researchers investigating the "predator model of PTSD." PMID- 21629857 TI - Information loss associated with imperfect observation and mismatched decoding. AB - We consider two types of causes leading to information loss when neural activities are passed and processed in the brain. One is responses of upstream neurons to stimuli being imperfectly observed by downstream neurons. The other is upstream neurons non-optimally decoding stimuli information contained in the activities of the downstream neurons. To investigate the importance of neural correlation in information processing in the brain, we specifically consider two situations. One is when neural responses are not simultaneously observed, i.e., neural correlation data is lost. This situation means that stimuli information is decoded without any specific assumption about neural correlations. The other is when stimuli information is decoded by a wrong statistical model where neural responses are assumed to be independent even when they are not. We provide the information geometric interpretation of these two types of information loss and clarify their relationship. We then concretely evaluate these types of information loss in some simple examples. Finally, we discuss use of these evaluations of information loss to elucidate the importance of correlation in neural information processing. PMID- 21629858 TI - Time scales of representation in the human brain: weighing past information to predict future events. AB - The estimates that humans make of statistical dependencies in the environment and therefore their representation of uncertainty crucially depend on the integration of data over time. As such, the extent to which past events are used to represent uncertainty has been postulated to vary over the cortex. For example, primary visual cortex responds to rapid perturbations in the environment, while frontal cortices involved in executive control encode the longer term contexts within which these perturbations occur. Here we tested whether primary and executive regions can be distinguished by the number of past observations they represent. This was based on a decay-dependent model that weights past observations from a Markov process and Bayesian Model Selection to test the prediction that neuronal responses are characterized by different decay half-lives depending on location in the brain. We show distributions of brain responses for short and long term decay functions in primary and secondary visual and frontal cortices, respectively. We found that visual and parietal responses are released from the burden of the past, enabling an agile response to fluctuations in events as they unfold. In contrast, frontal regions are more concerned with average trends over longer time scales within which local variations are embedded. Specifically, we provide evidence for a temporal gradient for representing context within the prefrontal cortex and possibly beyond to include primary sensory and association areas. PMID- 21629859 TI - Propagating waves in human motor cortex. AB - Previous studies in non-human primates (NHPs) have shown that beta oscillations (15-30 Hz) of local field potentials (LFPs) in the arm/hand areas of primary motor cortex (MI) propagate as traveling waves across the cortex. These waves exhibited two stereotypical features across animals and tasks: (1) The waves propagated in two dominant modal directions roughly 180 degrees apart, and (2) their propagation speed ranged from 10 to 35 cm/s. It is, however, unknown if such cortical waves occur in the human motor cortex. This study shows that the two properties of propagating beta waves are present in MI of a tetraplegic human patient while he was instructed to perform an instruction delay center-out task using a cursor controlled by the chin. Moreover, we show that beta waves are sustained and have similar properties whether the subject was engaged in the task or at rest. The directions of the successive sustained waves both in the human subject and a NHP subject tended to switch from one dominant mode to the other, and at least in the NHP subject the estimated distance traveled between successive waves traveling into and out of the central sulcus is consistent with the hypothesis of wave reflection between the border of motor and somatosensory cortices. Further, we show that the occurrence of the beta waves is not uniquely tied to periods of increased power in the beta frequency band. These results demonstrate that traveling beta waves in MI are a general phenomenon occurring in human as well as NHPs. Consistent with the NHP data, the dominant directions of the beta LFP waves in human aligned to the proximal to distal gradient of joint representations in MI somatotopy. This consistent finding of wave propagation may imply the existence of a hardwired organization of motor cortex that mediates this spatiotemporal pattern. PMID- 21629860 TI - Maximum probability reaction sequences in stochastic chemical kinetic systems. AB - The detailed behavior of many molecular processes in the cell, such as protein folding, protein complex assembly, and gene regulation, transcription and translation, can often be accurately captured by stochastic chemical kinetic models. We investigate a novel computational problem involving these models - that of finding the most-probable sequence of reactions that connects two or more states of the system observed at different times. We describe an efficient method for computing the probability of a given reaction sequence, but argue that computing most-probable reaction sequences is EXPSPACE-hard. We develop exact (exhaustive) and approximate algorithms for finding most-probable reaction sequences. We evaluate these methods on test problems relating to a recently proposed stochastic model of folding of the Trp-cage peptide. Our results provide new computational tools for analyzing stochastic chemical models, and demonstrate their utility in illuminating the behavior of real-world systems. PMID- 21629861 TI - The Involvement of Glutamate Metabolism in the Resistance to Thermal, Nutritional, and Oxidative Stress in Trypanosoma cruzi. AB - The inhibition of some glutamate metabolic pathways could lead to diminished parasite survival. In this study, the effects of L-methionine sulfoximine (MS), DL-methionine sulfone (MSO), and DL-methionine sulfoxide (MSE), three glutamate analogs, on several biological processes were evaluated. We found that these analogs inhibited the growth of epimastigotes cells and showed a synergistic effect with stress conditions such as temperature, nutritional starvation, and oxidative stress. The specific activity for the reductive amination of alpha ketoglutaric acid, catalyzed by the NADP(+)-linked glutamate dehydrogenase, showed an increase in the NADP(+) levels, when MS, MSE, and MSO were added. It suggests an eventual conversion of the compounds tested by the T. cruzi cells. The fact that trypomastigote bursting was not significantly inhibited when infected cells were treated with these compounds, remarks the existence of relevant metabolic differences among the different life-cycle stages. It must be considered when proposing a new therapeutic drug. PMID- 21629862 TI - Demonstrating Intertumoural Differences in Vascular-Metabolic Phenotype with Dynamic Contrast-Enhanced CT-PET. AB - Purpose. To assess whether the differences in vascular-metabolic relationships between lymphoma masses and colorectal liver metastases predicted from previous histopathological studies can be demonstrated by dynamic contrast-enhanced CT (DCE-CT) combined with fluorodeoxyglucose positron emission tomography (FDG-PET). Methods. DCE-CT and FDG-PET studies were drawn from an imaging archive for patients with either lymphoma masses (n = 11) or hepatic metastases from colorectal cancer (CRM: n = 12). Tumour vascularity was assessed using DCE-CT measurements of perfusion. Tumour glucose metabolism was expressed as the mean FDG Standardised Uptake Value (SUV(FDG)). The relationship between metabolism and vascularity in each group was assessed from SUV(FDG) /perfusion ratios and Pearson correlation coefficients. Results. An SUV(FDG) threshold of 3.0 was used to designate lymphoma masses as active (AL, n = 6) or inactive lymphoma (IL, n = 5). Tumour perfusion was significantly higher in AL (0.65 mL/min/mL) than CRM (0.37 mL/min/mL: P = .031) despite similar SUV(FDG) (5.05 and 5.33, resp.). AL demonstrated higher perfusion values than IL (0.24 mL/min/mL: P = .006). SUV(FDG)/perfusion was significantly higher in CRM (15.3 min) than IL (4.2 min, P < .01). There was no correlation between SUV(FDG) and perfusion for any patient group. PMID- 21629863 TI - Moving beyond the stigma: systematic review of video games and their potential to combat obesity. AB - Increasing epidemic proportions of overweight children in the United States presents formidable challenges for education and healthcare. Given the popularity and pervasiveness of video gaming culture in North American children, the perfect opportunity arises to investigate the potential of video games to promote healthful behaviour. Our objective was to systematically review the literature for possible benefits of active and educational video games targeting diet and physical activity in children. A review of English-language journal articles from 1998 to 2011 using EMBASE and PubMed was conducted. Thirty-four studies concerned with children, video games, physical, and/or nutritional outcomes were included. Results of these studies that showed some benefit (increased physical activity and nutritional knowledge as a result of gaming) demonstrate the possibility of video games to combat childhood obesity-looking beyond the stigma attached to gaming. PMID- 21629864 TI - Renal Sympathetic Denervation for the Treatment of Difficult-to-Control or Resistant Hypertension. AB - Hypertension represents a major health problem with an appalling annual toll. Despite the plethora of antihypertensive drugs, hypertension remains resistant in a considerable number of patients, thus creating the need for alternative strategies, including interventional approaches. Recently, catheter-based renal sympathetic denervation has been shown to be fairly safe and effective in patients with resistant hypertension. Pathophysiology of kidney function, interaction and crosstalk between the kidney and the brain, justifies the use of renal sympathetic denervation in the treatment of hypertension. Data from older studies have shown that sympathectomy has effectively lowered blood pressure and prolonged life expectancy of hypertensive patients, but at considerable cost. Renal sympathetic denervation is devoid of the adverse effects of surgical sympathectomy, due to its localized nature, is minimally invasive, and provides short procedural and recovery times. This paper outlines the pathophysiological background for renal sympathetic denervation, describes the past and the present of this interventional approach, and considers several future potential applications. PMID- 21629865 TI - Ambulatory blood pressure monitoring in resistant hypertension. AB - ABPM constitutes a valuable tool in the diagnosis of RH. The identification of white coat RH and masked hypertension (which may fulfill or not the definition of RH) is of great importance in the clinical management of such patients. Moreover, the various ABPM components such as average BP values, circadian BP variability patterns, and ambulatory BP-derived indices, such as ambulatory arterial stiffness index (AASI), add significantly to the risk stratification of RH. Lastly, ABPM may indicate the need for implementation of specific therapeutic strategies, such as chronotherapy, that is, administration-time dependent therapy, and the evaluation of their efficacy. PMID- 21629866 TI - A case of syncope induced in the supine position. AB - We experienced a reproducible supine syncope followed by upper abdominal pain. A 66-year-old man was transferred to our hospital after an episode of syncope during sleep. He had a history of acute pancreatitis, diabetes, hypertension, and dyslipidemia, but no history of presyncopal attack. One night, his wife noticed he was snoring abnormally in bed, and he did not respond to her voice until after she tried many times to wake him. The same attack was reproduced three times in the same situation. One of the attacks was recorded under a continuous ECG and radial tonometry. In this case, a presyncopal attack and a sense of ill-feeling were provoked by the patient lying in a prolonged supine position. He was eventually diagnosed as metastatic liver tumor 5 months after the first attack. Because few cases of syncopal attack have been reported in the supine position, its underlying mechanisms deserve consideration. PMID- 21629867 TI - Cell-phone use and self-reported hypertension: national health interview survey 2008. AB - Background. Cell-phone usage has increased dramatically over the last decade, along with a rising public concern over the health effects of using this device. The association between cell-phone usage and hypertension has not been examined before. Methods. We analysed data from 21,135 adults aged >=18 years who participated in the 2008 National Health Interview Survey. Based on reported cell phone use, participants were categorized as cell-phone nonusers, predominantly landline users, dual users of cell phone and landline, and predominantly cell phone users. The main outcome of interest was self-reported physician-diagnosed hypertension (n = 6,793). Results. 43.5% of the participants were cell-phone nonusers, while 13.8% were predominantly cell-phone users. We found that cell phone use was inversely associated with hypertension, independent of age, sex, race/ethnicity, smoking, alcohol consumption, education, body mass index (BMI), and physical activity. Compared to cell-phone nonusers, the multivariable odds ratio (95% confidence interval) of hypertension was 0.86 (0.75-0.98, P trend = .005) among predominantly cell-phone users. This inverse association between cell phone use and hypertension was stronger in women, those aged <60 years, whites, and those with BMI <25 kg/m(2). Conclusion. We found that cell-phone usage was protectively associated with self-reported hypertension in a nationally representative sample of US adults. PMID- 21629868 TI - Vagal withdrawal and sympathetic overactivity contribute to the genesis of early onset pregnancy-induced hypertension. AB - Objective. In this study, we have assessed sympathovagal imbalance (SVI) by spectral analysis of heart rate variability (HRV) that contributes to the genesis of early-onset PIH. Methods. Body mass index (BMI), basal heart rate (BHR), blood pressure (BP) and HRV indices such as LFnu, HFnu, LF-HF ratio, mean RR, SDNN and RMSSD were assessed in normal pregnant women (Control group) and pregnant women having risk factors for PIH (Study group) at all the trimesters pregnancy. Retrospectively, those who did not develop PIH (Study group I) were separated from those who developed PIH (Study group II). Study group II was subdivided into early-onset and late-onset PIH. Sympathovagal balance (LF-HF ratio) was correlated with BMI, BHR and BP. Results. LF-HF ratio was significantly high in study group II compared to study group I and control group, and in early-onset PIH group compared to the late-onset category at all the trimesters of pregnancy, which was significantly correlated with BHR and BP. Alteration in HFnu in early onset category was more prominent than the alteration in LFnu. Conclusion. Though the SVI in PIH is contributed by both sympathetic overactivity and vagal withdrawal, especially in early-onset type, SVI is mainly due to vagal inhibition. PMID- 21629869 TI - Long-term use of aldosterone-receptor antagonists in uncontrolled hypertension: a retrospective analysis. AB - Background. The long-term efficacy of aldosterone-receptor antagonists (ARAs) as add-on treatment in uncontrolled hypertension has not yet been reported. Methods. Data from 123 patients (21 with primary aldosteronism, 102 with essential hypertension) with difficult-to-treat hypertension who received an ARA between May 2005 and September 2009 were analyzed retrospectively for their blood pressure (BP) and biochemical response at first followup after start with ARA and the last follow-up available. Results. Systolic BP decreased by 22 +/- 20 and diastolic BP by 9.4 +/- 12 mmHg after a median treatment duration of 25 months. In patients that received treatment >5 years, SBP was 33 +/- 20 and DBP was 16 +/ 13 mmHg lower than at baseline. Multivariate analysis revealed that baseline BP and follow-up duration were positively correlated with BP response. Conclusion. Add-on ARA treatment in difficult-to-treat hypertension results in a profound and sustained BP reduction. PMID- 21629870 TI - Insulin resistance, obesity, hypertension, and renal sodium transport. AB - Sodium transport through various nephron segments is quite important in regulating sodium reabsorption and blood pressure. Among several regulators of this process, insulin acts on almost all the nephron segments and is a strong enhancer of sodium reabsorption. Sodium-proton exchanger type 3 (NHE3) is a main regulator of sodium reabsorption in the luminal side of proximal tubule. In the basolateral side of the proximal tubule, sodium-bicarbonate cotransporter (NBCe1) mediates sodium and bicarbonate exit from tubular cells. In the distal nephron and the connecting tubule, epithelial sodium channel (ENaC) is of great importance to sodium reabsorption. NHE3, NBCe1, and ENaC are all regulated by insulin. Recently with-no-lysine (WNK) kinases, responsible for familial hypertension, stimulating sodium reabsorption in the distal nephron, have been found to be also regulated by insulin. We will discuss the regulation of renal sodium transport by insulin and its roles in the pathogenesis of hypertension in insulin resistance. PMID- 21629871 TI - Management of hypertension and diabetes in obesity: non-pharmacological measures. AB - Obesity has become a global epidemic over the past few decades because of unhealthy dietary habits and reduced physical activity. Hypertension and diabetes are quite common among obese individuals and there is a linear relationship between the degree of obesity and these diseases. Lifestyle interventions like dietary modifications and regular exercise are still important and safe first line measures for treatment. Recently, bariatric surgery has emerged as an important and very effective treatment option for obese individuals especially in those with comorbidities like hypertension and diabetes. Though there are few effective drugs for the management of obesity, their efficacy is only modest, and they should always be combined with lifestyle interventions for optimal benefit. In this paper we aim to outline the non-pharmacological measures for the management of hypertension and diabetes in obesity. PMID- 21629872 TI - Circadian Variations in Blood Pressure, Heart Rate, and HR-BP Cross-Correlation Coefficient during Progression of Diabetes Mellitus in Rat. AB - Circadian changes in cardiovascular function during the progression of diabetes mellitus in the diabetes prone rat (BBDP) (n = 8) were studied. Age-matched diabetes-resistant rats (BBDR) served as controls. BP was recorded via telemetry in contiguous 4 hr time periods over 24 hours starting with 12 midnight to 4 am as period zero (P0). Prior to onset of diabetes BP was high at P0, peaked at P2, and then fell again at P3; BP and heart rate (HR) then increased gradually at P4 and leveled off at P5, thereby exhibiting a bipodal rhythm. These patterns changed during long-term diabetes. The cross-correlation coefficient of BP and HR was not significantly different across groups at onset, but it fell significantly at 9 months of duration of diabetes (BBDP: 0.39 +/- 0.06; BBDR: 0.65 +/- 0.03; P < .05). These results show that changes in circadian cardiovascular rhythms in diabetes mellitus became significant at the late stage of the disease. PMID- 21629873 TI - Prevalence of hypertension, obesity, diabetes, and metabolic syndrome in Nepal. AB - Background. This study was carried out to establish the prevalence of cardiovascular risks such as hypertension, obesity, and diabetes in Eastern Nepal. This study also establishes the prevalence of metabolic syndrome (MS) and its relationships to these cardiovascular risk factors and lifestyle. Methods. 14,425 subjects aged 20-100 (mean 41.4 +/- 15.1) were screened with a physical examination and blood tests. Both the International Diabetic Federation (IDF) and National Cholesterol Education Programme's (NCEP) definitions for MS were used and compared. Results. 34% of the participants had hypertension, and 6.3% were diabetic. 28% were overweight, and 32% were obese. 22.5% of the participants had metabolic syndrome based on IDF criteria and 20.7% according to the NCEP definition. Prevalence was higher in the less educated, people working at home, and females. There was no significant correlation between the participants' lifestyle factors and the prevalence of MS. Conclusion. The high incidence of dyslipidemia and abdominal obesity could be the major contributors to MS in Nepal. Education also appears to be related to the prevalence of MS. This study confirms the need to initiate appropriate treatment options for a condition which is highly prevalent in Eastern Nepal. PMID- 21629874 TI - Ambulatory blood pressure monitoring in diabetes and obesity-a review. AB - Diabetes mellitus and obesity are both related to the risk of cardiovascular disease and sudden death. In hypertensive guidelines, diabetes and obesity, especially abdominal obesity, are regarded as high-risk factors. Ambulatory blood pressure monitoring (ABPM) is an established method for the management of hypertension. However, ABPM is not a standard tool for the management of hypertension in diabetes and obesity. In this paper, recent data on the use of ABPM in diabetes and obesity will be discussed. In patients with diabetes, the ambulatory BP level has been shown to be better than clinic BP in predicting cardiovascular events. A riser pattern has been associated with increased risk of cardiovascular disease. White-coat hypertension and masked hypertension in diabetics constitute a moderate risk. A nondipping pattern is very common in obese hypertensive patients. In this paper, we will summarize the findings on the use of ABPM in patients with diabetes and obesity. PMID- 21629875 TI - Karolinska institutet 200-year anniversary. Symposium on traumatic injuries in the nervous system: injuries to the spinal cord and peripheral nervous system - injuries and repair, pain problems, lesions to brachial plexus. AB - The Karolinska Institutet 200-year anniversary symposium on injuries to the spinal cord and peripheral nervous system gathered expertise in the spinal cord, spinal nerve, and peripheral nerve injury field spanning from molecular prerequisites for nerve regeneration to clinical methods in nerve repair and rehabilitation. The topics presented at the meeting covered findings on adult neural stem cells that when transplanted to the hypoglossal nucleus in the rat could integrate with its host and promote neuron survival. Studies on vascularization after intraspinal replantation of ventral nerve roots and microarray studies in ventral root replantation as a tool for mapping of biological patterns typical for neuronal regeneration were discussed. Different immune molecules in neurons and glia and their very specific roles in synapse plasticity after injury were presented. Novel strategies in repair of injured peripheral nerves with ethyl-cyanoacrylate adhesive showed functional recovery comparable to that of conventional epineural sutures. Various aspects on surgical techniques which are available to improve function of the limb, once the nerve regeneration after brachial plexus lesions and repair has reached its limit were presented. Moreover, neurogenic pain after amputation and its treatment with mirror therapy were shown to be followed by dramatic decrease in phantom limb pain. Finally clinical experiences on surgical techniques to repair avulsed spinal nerve root and the motoric as well as sensoric regain of function were presented. PMID- 21629876 TI - Classification of intended phoneme production from chronic intracortical microelectrode recordings in speech-motor cortex. AB - We conducted a neurophysiological study of attempted speech production in a paralyzed human volunteer using chronic microelectrode recordings. The volunteer suffers from locked-in syndrome leaving him in a state of near-total paralysis, though he maintains good cognition and sensation. In this study, we investigated the feasibility of supervised classification techniques for prediction of intended phoneme production in the absence of any overt movements including speech. Such classification or decoding ability has the potential to greatly improve the quality-of-life of many people who are otherwise unable to speak by providing a direct communicative link to the general community. We examined the performance of three classifiers on a multi-class discrimination problem in which the items were 38 American English phonemes including monophthong and diphthong vowels and consonants. The three classifiers differed in performance, but averaged between 16 and 21% overall accuracy (chance-level is 1/38 or 2.6%). Further, the distribution of phonemes classified statistically above chance was non-uniform though 20 of 38 phonemes were classified with statistical significance for all three classifiers. These preliminary results suggest supervised classification techniques are capable of performing large scale multi class discrimination for attempted speech production and may provide the basis for future communication prostheses. PMID- 21629877 TI - Phytochemicals as Modulators of PPARs and RXRs. PMID- 21629878 TI - Maintaining optimal state probabilities in biological systems. AB - A biological problem is usually studied experimentally by reducing it into a number of modules. In contrast, the systems biology approach seeks to address the collective behavior of interacting molecules vis-a-vis the corresponding higher level behavior. Various attributes of a biological system are conditionally dependent on each other, and these conditionalities are usually represented through Bayesian networks for computing easily the joint probability for a state of an attribute. In this article, a genetic algorithm is investigated to a biological system, by representing it through a Bayesian network, for evaluating the optimum state probabilities of different attributes, in order to obtain a desired joint probability for a given state of an attribute. We believe that such a study would be helpful in achieving a desired health condition by maintaining various attributes of a system to their estimated optimum levels. PMID- 21629879 TI - Imaging brain tumor by dendrimer-based optical/paramagnetic nanoprobe across the blood-brain barrier. AB - A multimodal optical/paramagnetic nanoprobe, Den-Angio, was developed and demonstrated a capability to circumvent the blood brain barrier (BBB) and visualize brain tumors with high sensitivity in vivo. Den-Angio holds promise to pre-operatively localize brain tumors and make image-guided tumor resection possible during surgery. PMID- 21629880 TI - Multifunctional p-phosphonated calixarenes. AB - p-Phosphonic acid calix[n]arenes and their O-alkylated lower rim analogues are remarkably versatile macrocycles, with applications in selective diameter uptake of single walled carbon nano-tubes, as surfactants in stabilising and protecting nano-particles and graphene sheets, as crystal growth modifiers for inorganic systems, in encapsulating molecules of anti-cancer carboplatin, self assembly into nano-arrays, including nano-fibres and molecular capsules, and for binding metal ions including biologically relevant Ca(2+). They are readily accessible via five or six high yielding steps from the parent p-Bu(t) substituted compounds. PMID- 21629881 TI - Divergent reactions on racemic mixtures. AB - Methods that furnish enantioenriched products are crucial in modern organic synthesis. An underutilized strategy to arrive at enantioenriched products is to perform divergent reactions on racemic mixtures, where each enantiomer of the starting material reacts with a single chiral reagent to furnish two separable, non-enantiomeric products that are enantioenriched. Stereodivergent, regiodivergent and structurally divergent reactions on racemic mixtures are discussed in this tutorial review. PMID- 21629882 TI - AsS2Cl-an Arsenic(V) compound? Formation, stability and structure of gaseous AsSCl and AsS2Cl--a combined experimental and theoretical study. AB - By reaction of solid As(4)S(4) with gaseous Cl(2) at a temperature of 410 K gaseous AsSCl and AsS(2)Cl are formed. Unexpectedly in AsS(2)Cl the arsenic is not of formal oxidation state +V but +III: the molecular structure of AsS(2)Cl is arranged as a 1-chloro-dithia-arsirane and comprises an hitherto unknown AsS(2) three-membered ring. Thermodynamic data on AsSCl and AsS(2)Cl are obtained by mass spectrometry (MS). The experimental data are extended and confirmed by ab initio quantum chemical calculations (QC). The following values are given: Delta(f)H(0)(298)(AsSCl) = -5.2 kJ mol(-1) (MS), Delta(f)H(0)(298)(AsSCl) = 1.7 kJ mol(-1) (QC), S(0)(298)(AsSCl) = 296.9 J K(-1) mol(-1) (QC) and c(p)(0)(T)(AsSCl) = 55.77 + 3.97 * 10(-3)T- 4.38 * 10(5)T(-2)- 1.83 * 10(-6)T(2) and Delta(f)H(0)(298)(AsS(2)Cl) = -39.0 kJ mol(-1) (MS), Delta(f)H(0)(298)(AsS(2)Cl) = -20.2 kJ mol(-1) (QC), S(0)(298)(AsS(2)Cl) = 321.3 J K(-1) mol(-1) (QC) and c(p)(0)(T)(AsS(2)Cl) = 80.05 + 5.09 * 10(-3)T- 7.61 * 10(5)T(-2)- 2.35 * 10(-6)T(2) (298.15 K < T < 1000 K) (QC). The ionization energies are determined (IP(AsSCl) = 10.5, IP(AsS(2)Cl) = 10.2 eV). The IR spectrum of AsSCl is detected by means of matrix isolation spectroscopy. The estimated force constant f(As=S) = 4.47 mdyn.A(-1) gives rise to an As=S double bond. PMID- 21629883 TI - Palladium(IV) chemistry supported by pincer type ligands. AB - Palladium(IV) species supported by pincer type ligands that either generated in the stoichiometric reaction and were characterized by spectroscopic methods or were proposed in the catalysis, especially in Heck-Mizoroki, Suzuki-Miyaura and Negishi couplings, are summarized in this perspective. PMID- 21629884 TI - Large area flexible SERS active substrates using engineered nanostructures. AB - Surface enhanced Raman scattering (SERS) is an analytical sensing method that provides label-free detection, molecularly specific information, and extremely high sensitivity. The Raman enhancement that makes this method attractive is mainly attributed to the local amplification of the incident electromagnetic field that occurs when a surface plasmon mode is excited at a metallic nanostructure. Here, we present a simple, cost effective method for creating flexible, large area SERS-active substrates using a new technique we call shadow mask assisted evaporation (SMAE). The advantage of large, flexible SERS substrates such as these is they have more area for multiplexing and can be incorporated into irregular surfaces such as clothing. We demonstrate the formation of four different types of nanostructure arrays (pillar, nib, ellipsoidal cylinder, and triangular tip) by controlling the evaporation angle, substrate rotation, and deposition rate of metals onto anodized alumina nanoporous membranes as large as 27 mm. In addition, we present experimental results showing how a hybrid structure comprising of gold nanospheres embedded in a silver nano-pillar structure can be used to obtain a 50* SERS enhancement over the raw nanoparticles themselves. PMID- 21629885 TI - Beating cancer in multiple ways using nanogold. AB - Gold nanoparticles possess a unique combination of properties which allow them to act as highly multifunctional anti-cancer agents (X. H. Huang, P. K. Jain, I. H. El-Sayed and M. A. El-Sayed, Nanomedicine, 2007, 2, 681-693; P. Ghosh, G. Han, M. De, C. K. Kim and V. M. Rotello, Adv. Drug Delivery Rev., 2008, 60, 1307-1315; S. Lal, S. E. Clare and N. J. Halas, Acc. Chem. Res., 2008, 41, 1842-1851; D. A. Giljohann, D. S. Seferos, W. L. Daniel, M. D. Massich, P. C. Patel and C. A. Mirkin, Angew. Chem., Int. Ed., 2010, 49, 3280-3294). Not only can they be used as targeted contrast agents for photothermal cancer therapy, they can serve as scaffolds for increasingly potent cancer drug delivery, as transfection agents for selective gene therapy, and as intrinsic antineoplastic agents. This tutorial review will highlight some of the many forms and recent applications of these gold nanoparticle conjugates by our lab and others, as well as their rational design and physiologic interactions. PMID- 21629886 TI - Covalent immobilization of active lysozyme on Si/glass surface using alkoxy Fischer carbene complex on SAM. AB - A cross-metathesis reaction between an alkene terminated self-assembled monolayer (SAM) on glass/Si wafer and an alkene tethered Fischer carbene complex yielded a functionalized surface. Rapid aminolysis of the Fischer carbene moieties permit efficient anchoring of amine containing molecules on such a surface. Attachment of 1-pyrenemethylamine was thus monitored by ATR-IR spectroscopy and fluorescence microscopy. Similarly, BSA and lysozyme were individually grafted to such Fischer carbene modified surfaces using their pendant lysine residues. It has been demonstrated that the anchored lysozyme retains its bactericidal property. PMID- 21629887 TI - Hydrothermal synthesis of monodisperse single-crystalline alpha-quartz nanospheres. AB - Uniformly-sized, single-crystal alpha-quartz nanospheres have been synthesized at 200 degrees C and 15 atm under continuous stirring starting from uniform, amorphous Stober silica colloids and using NaCl and alkali hydroxide as mineralizers. Quartz nanosphere size is controlled by the colloid particle size via direct devitrification. Uniform, high-purity nanocrystalline quartz is important for understanding nanoparticle toxicology and for advanced polishing and nanocomposite fabrication. PMID- 21629888 TI - Activation of olefins with low-valent gallium compounds under ambient conditions. AB - The reactions of Ar'GaGaAr' (Ar' = C(6)H(3)-2,6-(C(6)H(3)-2,6-(i)Pr(2))(2)) with alkenes revealed the addition of two olefins per Ar'GaGaAr' under ambient conditions for ethylene, propene, 1-hexene and styrene but no reactions with more hindered or cyclic olefins. PMID- 21629889 TI - Effect of size of catalytically active phases in the dehydrogenation of alcohols and the challenging selective oxidation of hydrocarbons. AB - The size of the active phase is one of the most important factors in determining the catalytic behaviour of a heterogeneous catalyst. This Feature Article focuses on the size effects in two types of reactions, i.e., the metal nanoparticle catalysed dehydrogenation of alcohols and the metal oxide nanocluster-catalysed selective oxidation of hydrocarbons (including the selective oxidation of methane and ethane and the epoxidation of propylene). For Pd or Au nanoparticle-catalysed oxidative or non-oxidative dehydrogenation of alcohols, the size of metal nanoparticles mainly controls the catalytic activity by affecting the activation of reactants (either alcohol or O(2)). The size of oxidic molybdenum species loaded on SBA-15 determines not only the activity but also the selectivity of oxygenates in the selective oxidation of ethane; highly dispersed molybdenum species are suitable for acetaldehyde formation, while molybdenum oxide nanoparticles exhibit higher formaldehyde selectivity. Cu(II) and Fe(III) isolated on mesoporous silica are highly efficient for the selective oxidation of methane to formaldehyde, while the corresponding oxide clusters mainly catalyse the complete oxidation of methane. The lattice oxygen in iron or copper oxide clusters is responsible for the complete oxidation, while the isolated Cu(I) or Fe(II) generated during the reaction can activate molecular oxygen forming active oxygen species for the selective oxidation of methane. Highly dispersed Cu(I) and Fe(II) species also function for the epoxidation of propylene by O(2) and N(2)O, respectively. Alkali metal ions work as promoters for the epoxidation of propylene by enhancing the dispersion of copper or iron species and weakening the acidity. PMID- 21629890 TI - A highly symmetric diamond-like assembly of cyclotricatechylene-based tetrahedral cages. AB - The combination of six Cu(II) centres with four cyclotricatechylene (ctc(6-)) ligands generates large (Cu(6)ctc(4))(12-) cages with the topology of the tetrahedron. The anionic tetrahedral cages are cemented together, triangular face to triangular face, by Na(+) cations together with water molecules to generate a highly symmetric diamond-like assembly (space group Fd-3m, the same as that of diamond itself) whose adamantane-like units have an iodide ion at the centre of every cyclohexane-like face. PMID- 21629891 TI - Simplifying and improving the extraction of nitrate from freshwater for stable isotope analyses. AB - Determining the isotopic composition of nitrate (NO(3)(-)) in water can prove useful to identify NO(3)(-) sources and to understand its dynamics in aquatic systems. Among the procedures available, the 'ion-exchange resin method' involves extracting NO(3)(-) from freshwater and converting it into solid silver nitrate (AgNO(3)), which is then analysed for (15)N/(14)N and (18)O/(16)O ratios. This study describes a simplified methodology where water was not pre-treated to remove dissolved organic carbon (DOC) or barium cations (added to precipitate O bearing contaminants), which suited samples with high NO(3)(-) (>= 00 MUM or 25 mg L(-1) NO(3)(-)) and low DOC (typically < 17 MUM of C or 5 mg L(-1) C) levels. % N analysis revealed that a few AgNO(3) samples were of low purity (compared with expected % N of 8.2), highlighting the necessity to introduce quality control/quality assurance procedures for silver nitrate prepared from field water samples. Recommendations are then made to monitor % N together with % O (expected at 28.6, i.e. 3.5 fold % N) in AgNO(3) in order to better assess the type and gravity of the contamination as well as to identify potentially unreliable data. PMID- 21629892 TI - Mechanism of epoxide hydrolysis in microsolvated nucleotide bases adenine, guanine and cytosine: a DFT study. AB - Six water molecules have been used for microsolvation to outline a hydrogen bonded network around complexes of ethylene epoxide with nucleotide bases adenine (EAw), guanine (EGw) and cytosine (ECw). These models have been developed with the MPWB1K-PCM/6-311++G(3df,2p)//MPWB1K/6-31+G(d,p) level of DFT method and calculated S(N)2 type ring opening of the epoxide due to amino group of the nucleotide bases, viz. the N6 position of adenine, N2 position of guanine and N4 position of cytosine. Activation energy (E(act)) for the ring opening was found to be 28.06, 28.64, and 28.37 kcal mol(-1) respectively for EAw, EGw and ECw. If water molecules were not used, the reactions occurred at considerably high value of E(act), viz. 53.51 kcal mol(-1) for EA, 55.76 kcal mol(-1) for EG and 56.93 kcal mol(-1) for EC. The ring opening led to accumulation of negative charge on the developing alkoxide moiety and the water molecules around the charge localized regions showed strong hydrogen bond interactions to provide stability to the intermediate systems EAw-1, EGw-1 and ECw-1. This led to an easy migration of a proton from an activated water molecule to the alkoxide moiety to generate a hydroxide. Almost simultaneously, a proton transfer chain reaction occurred through the hydrogen bonded network of water molecules and resulted in the rupture of one of the N-H bonds of the quaternized amino group. The highest value of E(act) for the proton transfer step of the reaction was 2.17 kcal mol(-1) for EAw, 2.93 kcal mol(-1) for EGw and 0.02 kcal mol(-1) for ECw. Further, the overall reaction was exothermic by 17.99, 22.49 and 13.18 kcal mol(-1) for EAw, EGw and ECw, respectively, suggesting that the reaction is irreversible. Based on geometric features of the epoxide-nucleotide base complexes and the energetics, the highest reactivity is assigned for adenine followed by cytosine and guanine. Epoxide-mediated damage of DNA is reported in the literature and the present results suggest that hydrated DNA bases become highly S(N)2 active on epoxide systems and the occurrence of such reactions can inflict permanent damage to the DNA. PMID- 21629893 TI - Hyperbranched polymers versus dendrimers containing a carbosilane framework and terminal ammonium groups as antimicrobial agents. AB - A new family of amine- and ammonium-terminated hyperbranched polycarbosilanes (PCS) and dendrimers has been synthesized. The functionalization of a polycarbosilane matrix was carried out with peripheral allyl groups by two strategies in the case of PCS: 1) hydrosilylation of allyl amines with PCS containing terminal Si-H bonds, or 2) hydrosilylation of PCS-allyl with an aminosilane. Dendrimers with terminal amine groups were synthesized by hydrosilylation of allydimethylamine. Quaternized systems with MeI are soluble and stable in water or other protic solvent. The antibacterial properties of the ammonium-terminated hyperbranched polycarbosilanes and dendrimers have been evaluated showing that they act as potent biocides against Gram-positive and Gram negative bacterial strains. PMID- 21629894 TI - Chemical alterations to murine brain tissue induced by formalin fixation: implications for biospectroscopic imaging and mapping studies of disease pathogenesis. AB - Understanding biochemical mechanisms and changes associated with disease conditions and, therefore, development of improved clinical treatments, is relying increasingly on various biochemical mapping and imaging techniques on tissue sections. However, it is essential to be able to ascertain whether the sampling used provides the full biochemical information relevant to the disease and is free from artefacts. A multi-modal micro-spectroscopic approach, including FTIR imaging and PIXE elemental mapping, has been used to study the molecular and elemental profile within cryofixed and formalin-fixed murine brain tissue sections. The results provide strong evidence that amino acids, carbohydrates, lipids, phosphates, proteins and ions, such as Cl(-) and K(+), leach from tissue sections into the aqueous fixative medium during formalin fixation of the sections. Large changes in the concentrations and distributions of most of these components are also observed by washing in PBS even for short periods. The most likely source of the chemical species lost during fixation is the extra-cellular and intra-cellular fluid of tissues. The results highlight that, at best, analysis of formalin-fixed tissues gives only part of the complete biochemical "picture" of a tissue sample. Further, this investigation has highlighted that significant lipid peroxidation/oxidation may occur during formalin fixation and that the use of standard histological fixation reagents can result in significant and differential metal contamination of different regions of tissue sections. While a consistent and reproducible fixation method may be suitable for diagnostic purposes, the findings of this study strongly question the use of formalin fixation prior to spectroscopic studies of the molecular and elemental composition of biological samples, if the primary purpose is mechanistic studies of disease pathogenesis. PMID- 21629895 TI - Mechanistic investigations on the efficient catalytic decomposition of peroxynitrite by ebselen analogues. AB - In this study, ebselen and its analogues are shown to be catalysts for the decomposition of peroxynitrite (PN). This study suggests that the PN-scavenging ability of selenenyl amides can be enhanced by a suitable substitution at the phenyl ring in ebselen. Detailed mechanistic studies on the reactivity of ebselen and its analogues towards PN reveal that these compounds react directly with PN to generate highly unstable selenoxides that undergo a rapid hydrolysis to produce the corresponding seleninic acids. The selenoxides interact with nitrite more effectively than the corresponding seleninic acids to produce nitrate with the regeneration of the selenenyl amides. Therefore, the amount of nitrate formed in the reactions mainly depends on the stability of the selenoxides. Interestingly, substitution of an oxazoline moiety on the phenyl ring stabilizes the selenoxide, and therefore, enhances the isomerization of PN to nitrate. PMID- 21629896 TI - Facile synthesis of functional bismuth-amino acid coordination polymer nano structures. AB - Bismuth-asparagine coordination polymer nanostructures with diverse morphologies were fabricated by a facile aqueous bottom-up coordination-based self-assembly approach, and they exhibited unique properties in catalysis and biological aspects. PMID- 21629897 TI - Efficient access to a dihydropyran-containing macrolide via a transannular oxy Michael reaction: total synthesis of (+)-aspergillide C. AB - The second-generation enantioselective total synthesis of aspergillide C, a new type of 14-membered macrolide isolated from a marine-derived fungus, has been accomplished in a longest linear sequence of 15 steps from a chiral building block in 19% overall yield employing the 6-exo-trig transannular oxy-Michael reaction as the key step. PMID- 21629898 TI - A precise hexagonal octadecanuclear Ag macrocycle with significant luminescent properties. AB - The first hexagonal octadecanuclear Ag macrocycle [Ag(18)(TTTMB)(12)](NO(3))(18).30H(2)O has been hydrothermally synthesized. The outside and inner diameters of the unprecedented architecture are about 27.72(13) A and 15.37(7) A, respectively. This complex displays a huge increase in the intensity and lifetime of phosphorescence with decreasing temperature. PMID- 21629899 TI - Copper(II)-mediated chiral helicity amplification and inversion of meta ethynylpyridine polymers with metal coordination sites. AB - meta-Ethynylpyridine polymers bearing metal coordination sites associate with alkyl glycoside guests to show induced circular dichroism (ICD) bands. The addition of a Cu(II) ion changed the intensity or the sign of these ICD bands. The changes suggested chiral helicity amplification or inversion of the polymers by Cu(II)-mediated cross-linking at the coordinating side chains. PMID- 21629900 TI - Efficient catalytic decomposition of sulfuric acid with copper vanadates as an oxygen-generating reaction for solar thermochemical water splitting cycles. AB - Copper pyrovanadate (Cu(2)V(2)O(7)) was found for the first time to achieve the high catalytic activity for the decomposition of sulfuric acid to evolve O(2) at moderate temperatures around 650 degrees C, which is essential for the development of solar thermochemical water splitting cycles. PMID- 21629901 TI - TiO(2)@carbon core/shell nanofibers: controllable preparation and enhanced visible photocatalytic properties. AB - TiO(2)@carbon core/shell nanofibers (TiO(2)@C NFs) with different thinkness of carbon layers (from 2 to 8 nm) were fabricated by combining the electrospinning technique and hydrothermal method. The results showed that a uniform graphite carbon layer was formed around the electrospun TiO(2) nanofiber via C-O-Ti bonds. By adjusting the hydrothermal fabrication parameters, the thickness of carbon layer could be easily controlled. Furthermore, the TiO(2)@C NFs had remarkable light absorption in the visible region. The photocatalytic studies revealed that the TiO(2)@C NFs exhibited enhanced photocatalytic efficiency of photodegradation of Rhodamine B (RB) compared with the pure TiO(2) nanofibers under visible light irradiation, which might be attributed to high separation efficiency of photogenerated electrons and holes based on the synergistic effect between carbon as a sensitizer and TiO(2) with one dimension structure. Notably, the TiO(2)@C NFs could be easily recycled due to their one-dimensional nanostructural property. PMID- 21629902 TI - Phosphine-catalyzed [3 + 2] cycloaddition of allenoates with trifluoromethylketones: synthesis of dihydrofurans and tetrahydrofurans. AB - The triphenylphosphine-catalyzed formal [3 + 2] cycloaddition of allenoates and trifluoromethylketones was realized to give the corresponding dihydrofurans in good yields with excellent gamma-regioselectivities. Hydrogenation of the dihydrofurans gave 2,4,4-trisubstituted tetrahydrofurans in good yields with exclusive cis-selectivities. PMID- 21629903 TI - Structural organization and phase behaviour of 1-butyl-3-methylimidazolium hexafluorophosphate: an high pressure Raman spectroscopy study. AB - The complexity of the phase diagram of a representative room temperature ionic liquid (1-butyl-3-methylimidazolium hexafluorophosphate, [bmim][PF(6)]) is explored by means of Raman spectroscopy at high pressure (up to 1000 MPa) and high temperature (from room temperature to 100 degrees C) conditions. The first experimental evidence of the existence of a second crystalline phase for this salt at high pressure conditions is provided. By comparing the low frequency vibrational bands for the liquid state and the two observed crystalline phases, we confirm the scenario that considers the crystal polymorphism in this class of materials as a consequence of the rotational isomerism of the butyl chain. Furthermore the pressure dependence of other vibrational bands indicates the existence of a structural rearrangement across p~ 50 MPa at ambient temperature. PMID- 21629904 TI - Aerobic, Cu-catalyzed desulfitative C-C bond-forming reaction of ketene dithioacetals/vinylogous thioesters and arylboronic acids. AB - A new Cu-catalyzed thioorganic-boronic acid desulfitative C-C bond-forming reaction involving ketene dithioacetals/ vinylogous thioesters is reported to proceed without the assistance of ligating S-pendant. Vinylogous thiolesters and tetrasubstituted olefins were prepared by this reaction in which Cu catalyst plays a dual role under aerobic conditions. PMID- 21629905 TI - Novel crosslinked membranes based on sulfonated poly(ether ether ketone) for direct methanol fuel cells. AB - Novel covalently crosslinked membranes based on sulfonated poly(ether ether ketone) and carboxylated polysulfone exhibit much lower methanol crossover and better performance in direct methanol fuel cells at 65 degrees C in 1 and 2 M methanol solutions compared to Nafion 115 membranes. PMID- 21629906 TI - On the utility of S-mesitylsulfinimines for the stereoselective synthesis of chiral amines and aziridines. AB - The synthetic utility of S-mesitylsulfinimines for the synthesis of chiral amines and aziridines was examined through their reactions with Grignard reagents, with the ylides derived from trimethylsulfonium iodide and S-allyl tetrahydrothiophenium bromide and through an aza-Darzens manifold, affording convenient access to a diverse range of highly substituted chiral amines and aziridines in high yields and excellent stereoselectivities. PMID- 21629907 TI - Selective visualization of point defects in carbon nanotubes at the atomic scale by an electron-donating molecular tip. AB - Electron-donating molecular tips were used for the observation of single-walled carbon nanotubes (SWNTs). Defects in SWNTs were selectively visualized at the atomic scale on the basis of charge-transfer interaction with the molecular tip. PMID- 21629908 TI - A new donor-acceptor molecule with uniaxial anisotropy for efficient vacuum deposited organic solar cells. AB - A new D-pi-A molecule (TPDCDTS) adopting coplanar diphenyl-substituted dithienosilole as a central pi-bridge of triphenylamine (donor) and dicyanovinylene (acceptor) has been synthesized as donor material for small molecule organic solar cells (SMOSC) incorporating C(70) as an acceptor showed an appreciable power conversion efficiency of 3.82%. PMID- 21629909 TI - DNAzyme crosslinked hydrogel: a new platform for visual detection of metal ions. AB - We propose the use of DNAzyme as a crosslinker of hydrogel to develop a catalytic platform for the sensing of metal ions. The DNAzyme crosslinked hydrogel can undergo gel-sol transition in response to Cu(2+) ions, which enables sensitive visual detection of Cu(2+) by observing the release of pre-trapped AuNPs. PMID- 21629910 TI - Biocatalytic synthesis of gold nanoparticles with cofactor regeneration in recombinant Escherichia coli cells. AB - Here we report the enzymatic synthesis of gold nanoparticles (Au NPs) by an engineered Escherichia coli harboring an NADH cofactor regeneration system coupled with glycerol dehydrogenase, which can be triggered by the addition of exogenous glycerol. PMID- 21629911 TI - Stabilization and functionalization of iron oxide nanoparticles for biomedical applications. AB - Superparamagnetic iron oxide nanoparticles (NPs) are used in a rapidly expanding number of research and practical applications in the biomedical field, including magnetic cell labeling separation and tracking, for therapeutic purposes in hyperthermia and drug delivery, and for diagnostic purposes, e.g., as contrast agents for magnetic resonance imaging. These applications require good NP stability at physiological conditions, close control over NP size and controlled surface presentation of functionalities. This review is focused on different aspects of the stability of superparamagnetic iron oxide NPs, from its practical definition to its implementation by molecular design of the dispersant shell around the iron oxide core and further on to its influence on the magnetic properties of the superparamagnetic iron oxide NPs. Special attention is given to the selection of molecular anchors for the dispersant shell, because of their importance to ensure colloidal and functional stability of sterically stabilized superparamagnetic iron oxide NPs. We further detail how dispersants have been optimized to gain close control over iron oxide NP stability, size and functionalities by independently considering the influences of anchors and the attached sterically repulsive polymer brushes. A critical evaluation of different strategies to stabilize and functionalize core-shell superparamagnetic iron oxide NPs as well as a brief introduction to characterization methods to compare those strategies is given. PMID- 21629912 TI - Effect of peptide-based captopril analogues on angiotensin converting enzyme activity and peroxynitrite-mediated tyrosine nitration. AB - Angiotensin converting enzyme (ACE) regulates the blood pressure by converting angiotensin I to angiotensin II and bradykinin to bradykinin 1-7. These two reactions elevate the blood pressure as angiotensin II and bradykinin are vasoconstrictory and vasodilatory hormones, respectively. Therefore, inhibition of ACE is an important strategy for the treatment of hypertension. The natural substrates of ACE, i.e., angiotensin II and bradykinin, contain a Pro-Phe motif near the site of hydrolysis. Therefore, there may be a Pro-Phe binding pocket at the active site of ACE, which may facilitate the substrate binding. In view of this, we have synthesized a series of thiol- and selenol-containing dipeptides and captopril analogues and studied their ACE inhibition activities. This study reveals that both the selenol or thiol moiety and proline residues are essential for ACE inhibition. Although the introduction of a Phe residue to captopril and its selenium analogue considerably reduces the inhibitory effect, there appears to be a Phe binding pocket at the active site of ACE. PMID- 21629913 TI - Rearrangement of lithiated S-alkyl O,O-dialkyl thiophosphates: scope and stereochemistry of the thiophosphate-mercaptophosphonate rearrangement. AB - S-Alkyl O,O-dialkyl thiophosphates are prepared by alkylation of the triethylammonium salt of O,O-diisopropyl thiophosphoric acid. S-Benzyl thiophosphate was metallated at temperatures of >=-45 degrees C by trityllithium and LiTMP (lithium 2,2,6,6-tetramethylpiperidide) and S-alkyl thiophosphates only by LiTMP to give dipole-stabilised carbanions which rearrange to alpha mercaptophosphonates in yields of up to 45%. Metallation occurs with a high primary kinetic isotope effect (k(H)/k(D) up to ~50). When the lithium (R)-N isopropyl-1-phenylethylamide was used to induce the isomerisation of S-pentyl thiophosphate an alpha-mercaptophosphonate with an ee of 22% was isolated. (R)-S [1-D(1)]hexyl O,O-diisopropyl thiophosphate was rearranged to a dextrorotary alpha-mercapto-[1-D(1)]hexylphosphonate, whose (R)-configuration was determined by chemical correlation. The thiophosphate-mercaptophosphonate rearrangement proceeds with retention of configuration. PMID- 21629914 TI - Debundling, selection and release of SWNTs using fluorene-based photocleavable polymers. AB - Photocleavable polymers based on 9,9-dialkylfluorene backbone and o nitrobenzylether were designed and synthesized to obtain stable (n,m) enriched suspensions of semiconducting SWNTs in toluene. Photoirradiation of the suspensions triggered the precipitation of the SWNTs and TEM images indicate close packing of SWNTs pointing at partial removal of the coating polymer. PMID- 21629915 TI - Highly stable surface modifications of poly(3-caprolactone) (PCL) films by molecular self-assembly to promote cells adhesion and proliferation. AB - In this paper, we report a simple and versatile surface coating method to functionalize poly(3-caprolactone) (PCL) films by molecular self-assembly of a hydrogelator. PMID- 21629916 TI - Three-dimensionally ordered mesoporous Pd networks templated by a silica super crystal and their application in formic acid electrooxidation. AB - Three-dimensionally ordered mesoporous Pd networks fabricated by a simple reduction method in solution using a face centered cubic silica super crystal as template exhibit high electroactivity in formic acid oxidation. PMID- 21629917 TI - Deposition of platinum patterns by a liquid process. AB - In contrast to the traditional chemical vapor deposition technique under high vacuum, we introduce a deposition method in liquid to prepare Pt patterns on substrate near 100 degrees C by seed growth. PMID- 21629918 TI - Mesoporous MFI zeolites by microwave induced assembly between sulfonic acid functionalized MFI zeolite nanoparticles and alkyltrimethylammonium cationic surfactants. AB - Mesoporous MFI zeolites (ZSM-5, TS-1, S-1) having intracrystalline mesoporosity within zeolite crystals were synthesized by microwave induced assembly through the ionic interaction between the sulfonic acid functionalized MFI zeolite nanoparticles and alkyltrimethylammonium cationic surfactants. PMID- 21629919 TI - Promoted aerobic oxidation of benzyl alcohol on CNT supported platinum by iron oxide. AB - The catalytic activity of Pt/CNT for benzyl alcohol aerobic oxidation was remarkably improved by decorating iron oxide on Pt nanoparticles, and electrochemical measurements evidenced the enhanced activation of oxygen and benzyl alcohol at the FeO(x)/Pt interface. PMID- 21629920 TI - Integrating top-down and self-assembly in the fabrication of peptide and protein based biomedical materials. AB - The capacity to create an increasing variety of bioactive molecules that are designed to assemble in specific configurations has opened up tremendous possibilities in the design of materials with an unprecedented level of control and functionality. A particular challenge involves guiding such self-assembling interactions across scales, thus precisely positioning individual molecules within well-organized, highly-ordered structures. Such hierarchical control is essential if peptides and proteins are to serve as both structural and functional building blocks of biomedical materials. To achieve this goal, top-down techniques are increasingly being used in combination with self-assembling systems to reproducibly manipulate, localize, orient and assemble peptides and proteins to form organized structures. In this tutorial review we provide insight into how both standard and novel top-down techniques are being used in combination with peptide or protein self-assembly to create a new generation of functional materials. PMID- 21629921 TI - A significant role of alkaline cations on the Reimer-Tiemann reaction. AB - The Reimer-Tiemann (R-T) reaction was investigated by DFT calculations. A model composed of CHCl(3), PhO(-)(Na(+))H(2)O and [NaOH(H(2)O)(2)](2) was employed for geometry optimizations. A K(+)-containing model was also investigated. The dichlorocarbene reagent, which has been thought of for a long time, was found to intervene only transiently in the carbenoid form. In this form, the Na(+) (or K(+)) coordination to CCl(2) enhances its electrophilicity toward C(6)H(5)O(-). The counter ion also works to stabilize the precursor phenoxide ion and intermediates of the substituted phenoxides in the hexagonal pyramidal coordination. The Na(+)-containing reaction consists of seven elementary processes, (K(+), six ones) with extremely high exothermicity and spontaneity. PMID- 21629922 TI - 1,3-Diphenylbenzo[e][1,2,4]triazin-7(1H)-one: selected chemistry at the C-6, C-7 and C-8 positions. AB - 1,3-Diphenylbenzo[e][1,2,4]triazin-7(1H)-one (6) reacts with tetracyanoethylene (TCNE) or tetracyanoethylene oxide (TCNEO) to give the deep green 2-[1,3 diphenylbenzo[e][1,2,4]triazin-7(1H)-ylidene]propanedinitrile (11) in 17 and 15% yields, respectively. Nucleophiles such as amines, alkoxides, thiols and Grignard reagents all reacted with the 1,3-diphenylbenzotriazinone 6 regioselectively at C 6, while halogenating agents reacted exclusively at C-8. Furthermore, 8-iodo-1,3 diphenylbenzo[e][1,2,4]triazin-7(1H)-one (32) undergoes palladium-catalysed Suzuki-Miyaura and Stille coupling reactions to give 8-aryl- or heteroaryl substituted benzotriazinones. By combining both the C-6 and C-8 chemistries 1,3,6,8-tetraphenylbenzo[e][1,2,4]triazin-7(1H)-one (42) and 1,3-diphenyl-6,8 di(thien-2-yl)-benzo[e][1,2,4]triazin-7(1H)-one (43) can be prepared. All new compounds are fully characterized. PMID- 21629923 TI - Amperometric detection of antibodies in serum: performance of self-assembled cyclodextrin/cellulose polymer interfaces as antigen carriers. AB - A bifunctionalised carboxymethyl-cellulose polymer bearing adamantane units and an antigenic fragment forms a highly stable interfacial complex with a betaCD containing surface. This allows the highly sensitive detection of antibodies using an amperometric immunosensor. PMID- 21629924 TI - Conia-ene annulation of the alpha-cyano beta-TMS-capped alkynyl cycloalkanone system and its synthetic application. AB - Under catalysis with ZnI(2), an effective annulation process of omega silylacetylenic alpha-cyano ketones, leading to the formation of various bicyclic frameworks characterized with a TMS-containing methylenecyclopentane ring, has been developed. PMID- 21629937 TI - To understand the whole, you must know the parts: unraveling the roles of protein DNA interactions in genome regulation. AB - The regulation of gene transcription is fundamental to the existence of complex multicellular organisms such as humans. This process dictates which genes are expressed in which tissues, and controls how various cell types grow, differentiate, and respond to their environments. Although the deciphering of the human genome sequence has given us the "source code" for life, we still know far too little about the mechanisms that control which sets of genes are active in which tissues, and how their expression is regulated. It is clear, however, that much of this system depends upon the sequence-specific interactions of regulatory proteins with particular genetic loci. To be able to unravel the details of these interactions on a genome-wide basis, it is necessary to know what proteins are bound to the DNA where in the genome, and to be able to monitor how those proteins change over time and in response to external stimuli. Developing a new technology to provide this information constitutes a "Grand Challenge" for Analytical Chemistry. In this brief article we outline the nature of this challenge, and propose one strategy to address it. PMID- 21629938 TI - 4,6-Di-O-pentyl-3-O-trifluoroacetyl/propionyl cyclofructan stationary phases for gas chromatographic enantiomeric separations. AB - 4,6-Di-O-pentyl-3-O-trifluoroacetyl cycloinulohexaose (DP-TA-CF6) and 4,6-di-O pentyl-3-O-propionyl cycloinulohexaose (DP-PN-CF6) were synthesized and used as chiral stationary phases (CSPs) in gas chromatography (GC). The chiral recognition ability of the two CSPs was investigated. A total of 47 racemic compounds were separated on the two new CSPs, including derivatized amino acids, amino alcohols, amines, alcohols, tartrates and lactones. Interestingly, several analytes were only separated on either the DP-TA-CF6 or the DP-PN-CF6 phase. The chiral recognition mechanism was evaluated through thermodynamic analysis. The result indicated there was no inclusion complex formation involved in the chiral recognition process. PMID- 21629939 TI - Luminescence enhancement and tuning via multiple cooperative supramolecular interactions in an ion-paired multinuclear complex. AB - [(3,5-(CF(3))(2)Pz)(AgL)(2)](+)[Ag(5)(3,5-(CF(3))(2)Pz)(6)(CH(3)CN)](-) (L = 2 (N,N-diethylanilino-4-yl)-4,6-bis(3,5-dimethylpyrazol-1-yl)-1,3,5-triazine) shows bright and tunable emissions influenced by its supramolecular structure. Columnar stacks are assembled via cooperative interactions that include Ag(I)...Ag(I) argentophilic bonding, pi...pi stacking and Ag(I)...pi interactions. PMID- 21629940 TI - A transparent photo-responsive organogel based on a glycoluril supergelator. AB - The synthesis of an azo-benzene glycoluril supergelator is reported. The control over the gel/sol state can be accomplished by irradiation and heat, but also (in a unidirectional sense) by incorporation of the glycoluril into a more stable supramolecular assembly. PMID- 21629941 TI - Facile synthesis of spiny mesoporous titania tubes with enhanced photocatalytic activity. AB - We demonstrate here for the first time an original and facile approach to synthesizing a novel TiO(2) tube with a spiny mesoporous wall. The photocatalytic activity of the spiny mesoporous tube was greatly enhanced owing to the superior light reflecting ability of the tubular structure as well as the crosslinked spiny nanosheets. PMID- 21629942 TI - Balancing binding strength and charge transfer lifetime in supramolecular associates of fullerenes. AB - A macrocyclic exTTF host for fullerenes offers control over the electronic coupling between an electron donor and an acceptor, and stabilizes the charge separated state lifetimes into the range of 500 ps. PMID- 21629943 TI - Artificial luminescent protein as a bioprobe for time-gated luminescence bioimaging. AB - An artificial luminescent protein, apoferritin-encapsulated luminescent europium complex, has been designed/fabricated and displays good biocompatibility and long lived luminescence, which means it can be used as a bioprobe to image living cells with a time-gated mode. PMID- 21629944 TI - DNA-templated ensemble for label-free and real-time fluorescence turn-on detection of enzymatic/oxidative cleavage of single-stranded DNA. AB - We have demonstrated a DNA-templated small molecule ensemble that allows the label-free fluorescence turn-on detection of enzymatic or oxidative cleavage of single-stranded DNA with high sensitivity and selectivity. PMID- 21629945 TI - Synthesis and enantioselective hydrogenation of seven-membered cyclic imines: substituted dibenzo[b,f][1,4]oxazepines. AB - Highly enantioselective hydrogenation of seven-membered cyclic imines, substituted dibenzo[b,f][1,4]oxazepines, was achieved, with up to 94% ee, by using the [Ir(COD)Cl](2)/(S)-Xyl-C(3)*-TunePhos complex as the catalyst in the presence of morpholine-HCl. PMID- 21629946 TI - Modification of the sigma-framework of [60]fullerene for bulk-heterojunction solar cells. AB - Modification of the sigma-framework of [60]fullerene has been shown to be a new avenue toward n-type materials for bulk-heterojunction solar cells. This approach allowed distinct control of the LUMO level of the C(60)pi-system and resulted in a clear improvement of an open-circuit voltage. PMID- 21629947 TI - Modular ligand variation in calcium bisimidazoline complexes: effects on ligand redistribution and hydroamination catalysis. AB - A series of calcium complexes supported by chiral bisimidazoline ligands have been studied in the catalytic intramolecular hydroamination/cyclisation of amino olefins. The complexes [Ca(R-BIM){N(SiMe(3))(2)}(THF)] (R = 4-C(6)H(4)Me, 5a, 4 C(6)H(4)F, 5b and (t)Bu, 5c) have given competitive enantioselectivities (up to 12%) when compared to current literature studies involving calcium. Bisimidazolines offer a significant advance over similar bisoxazoline ligands, by allowing a greater structural variance through a modular synthetic pathway. PMID- 21629948 TI - Dielectrophoresis-based cellular microarray chip for anticancer drug screening in perfusion microenvironments. AB - We present a dielectrophoresis (DEP)-based cellular microarray chip for cell based anticancer drug screening in perfusion microenvironments. Human breast cancer cells, MCF7, were seeded into the chip and patterned via DEP forces onto the planar interdigitated ring electrode (PIRE) arrays. Roughly, only one third of the cell amount was required for the chip compared to that for a 96-well plate control. Drug concentrations (cisplatin or docetaxel) were stably generated by functional integration of a concentration gradient generator (CGG) and an anti crosstalk valve (ACV) to treat cells for 24 hours. Cell viability was quantified using a dual staining method. Results of cell patterning show substantial uniformity of patterned cells (92 +/- 5 cells per PIRE). Furthermore, after 24 hour drug perfusion, no statistical significance in dose-responses between the chip and the 96-well plate controls was found. The IC(50) value from the chip also concurred with the values from the literature. Moreover, the perfusion culture exhibited reproducibility of drug responses of cells on different PIREs in the same chamber. The chip would enable applications where cells are of limited supply, and supplement microfluidic perfusion cultures for clinical practices. PMID- 21629949 TI - Reactions in double emulsions by flow-controlled coalescence of encapsulated drops. AB - We demonstrate a microfluidic method to first generate double emulsion droplets containing two different inner drops, and to then control the internal coalescence of the encapsulated drops. The advantages of the core-coalescence method are illustrated by fabricating high viscosity particles and by demonstrating the dissolution of cell membranes. PMID- 21629950 TI - Biocompatible multi-address 3D cell assembly in microfluidic devices using spatially programmable gel formation. AB - Programmable 3D cell assembly under physiological pH conditions is achieved using electrodeposited stimuli-responsive alginate gels in a microfluidic device, with parallel sidewall electrodes enabling direct observation of the cell assembly. Electrically triggered assembly and subsequent viability of mammalian cells is demonstrated, along with spatially programmable, multi-address assembly of different strains of E. coli cells. Our approach enables in vitro study of dynamic cellular and inter-cellular processes, from cell growth and stimulus/response to inter-colony and inter-species signaling. PMID- 21629951 TI - Famous last words... PMID- 21629952 TI - Are we ready for industrial production of bioactive plant secondary metabolites utilizing endophytes? AB - The prospect of endophytes capable of producing associated plant natural products with therapeutic potential is undeniable. However, the unyielding struggle so far to transform these potential sources into sustainable microbial manufacturers on a commercial scale has been disappointing. In this Viewpoint, we address this trade-off between endophyte discovery and viable industrial application. PMID- 21629953 TI - Solid phase combinatorial synthesis of a xanthone library using click chemistry and its application to an embryonic stem cell probe. AB - We report the first solid phase synthesis of a xanthone library CX and its application to embryonic stem cell probe development. The CX library was further derivatised with an activated ester resin to provide an acetylated CX (CXAC) library. Screening of these libraries led to the discovery of a novel fluorescent mESC probe, CDb8. PMID- 21629954 TI - Identification of secondary effects of hyperexcitability by proteomic profiling of myotonic mouse muscle. AB - Myotonia is a symptom of various genetic and acquired skeletal muscular disorders and is characterized by hyperexcitability of the sarcolemma. Here, we have performed a comparative proteomic study of the genetic mouse models ADR, MTO and MTO*5J of human congenital myotonia in order to determine myotonia-specific changes in the global protein complement of gastrocnemius muscle. Proteomic analyses of myotonia in the mouse, which is caused by mutations in the gene encoding the muscular chloride channel Clc1, revealed a generally perturbed protein expression pattern in severely affected ADR and MTO muscle, but less pronounced alterations in mildly diseased MTO*5J mice. Alterations were found in major metabolic pathways, the contractile machinery, ion handling elements, the cellular stress response and cell signaling mechanisms, clearly confirming a glycolytic-to-oxidative transformation process in myotonic fast muscle. In the long-term, a detailed biomarker signature of myotonia will improve our understanding of the pathobiochemical processes underlying this disorder and be helpful in determining how a single mutation in a tissue-specific gene can trigger severe downstream effects on the expression levels of a very large number of genes in contractile tissues. PMID- 21629955 TI - Confirmation of the classic tris(2,2'-bipyridyl)ruthenium(II) and oxalate electrochemiluminescence mechanism using EPR spectroscopy. AB - A chemically initiated adaptation of the classic [Ru(bipy)(3)](2+)/oxalate electrochemiluminescence coreactant system has revealed the elusive radical intermediates of the light-producing pathway. Oxalyl (HC(2)O(4)) and hydroxyformyl (HCO(2)) radicals have been captured on a quartz surface and characterised using EPR spectroscopy. PMID- 21629956 TI - pH responsive self-healing hydrogels formed by boronate-catechol complexation. AB - Here we report the synthesis and characterization of pH-responsive, self-healing hydrogels based on boronate-catechol complexation. PMID- 21629957 TI - Fluorescent and luminescent probes for detection of reactive oxygen and nitrogen species. AB - Oxidative and nitrosative stress induced by ROS/RNS play crucial roles in a wide range of physiological processes and are also implicated in various diseases, including cancer and neurodegenerative disorders. Sensitive and selective methods for the detection of ROS/RNS based on fluorescent and luminescent probes are of great use in monitoring the in vivo production of these species and elucidating their biological functions. This critical review highlights recent advances that have been made in the development of fluorescent and luminescent probes employed to monitor various ROS/RNS (132 references). PMID- 21629958 TI - Enthalpy-entropy correlations as chemical guides to unravel self-assembly processes. AB - Intermolecular connections play a crucial role in biology (recognition, signalling, binding), in physics (material cohesion) and in chemistry ((supra)molecular engineering). While a phenomenological thermodynamic free energy approach for modelling self-assemblies is now at hand, a more satisfying description based on the chemically-intuitive enthalpic and entropic contributions remains elusive. On the other hand, the innumerable reports of empirical enthalpy/entropy correlations characterizing intermolecular interactions justify a questioning about the emergence and exploitation of an apparent 'fourth law of thermodynamics', which could provide a simple manipulation of intermolecular binding processes. This tutorial Perspective aims at highlighting the current level of non-quantum rationalization of enthalpy entropy correlations and their chemical consequences on the tuning and on the programming of intermolecular interactions in pure materials, and in diluted solutions. PMID- 21629959 TI - Synthesis, characterization, mechanochromism and photochromism of [Fe(dm4bt)3][FeCl4]2 and [Fe(dm4bt)3][FeBr4]2, along with the investigation of steric influence on spin state. AB - The complexes [Fe(dm4bt)(3)][FeCl(4)](2) (1) and [Fe(dm4bt)(3)][FeBr(4)](2) (2) were prepared from the reaction of 2,2'-dimethyl-4,4'-bithiazole (dm4bt) with FeCl(3).6H(2)O and FeBr(3), respectively, in methanol. Both complexes were characterized by IR, UV-Vis and (1)H NMR spectroscopy and their structures were studied by single-crystal diffraction. The methylated bithiazole led to high spin Fe(II) centers in the octahedral cation part of complexes 1 and 2 with Fe-N distance of 2.220 A, in spite of the low spin octahedral Fe(II) complexes with unsubstituted bithiazole ligands. Crystal structure determination of 2 was performed at 90, 120 and 298 K. Temperature reduction to 90 K resulted in a decrease in the Fe-N bond length to 2.206 A which is still in the range of high spin Fe(II). Complex 1 shows a reversible mechanochromic effect from the crystalline phase to powder form from red to yellow; it also displays reversible photochromism from yellow to green in solution under sunlight. The magnetic behaviour of the complexes was also studied at 2-300 K. The temperature dependence of chi(m)T curves for the two forms of 1, crystal and powder, demands some changes in their magnetic behavior, causing different colors i.e. red and yellow. At low temperatures, chi(m)T decreases where the decrease starts at around 65 K for compound 1, and at around 100 K for compound 2, due to different counterions. The two complexes exhibit antiferromagnetism at around 4 K. PMID- 21629960 TI - Toward design of multiple-property inorganic-organic hybrid compounds based on face-sharing octahedral iodoplumbate chains. AB - In this review article, we have illustrated the strategies developed to achieve inorganic-organic hybrid compounds with technologically important physical properties. A series of target inorganic-organic hybrid compounds have been accomplished by incorporating the functional organic components (with a large hyperpolarizability and luminophore Schiff base cation) into the highly polarizable one-dimensional (1-D) iodoplumbate chain network. The effect of substituent features in the phenyl ring of the Schiff base cation on its molecular conformation as well as the crystal packing structure of the hybrid compound will be discussed and the multiple physical properties (ferroelectricity, NLO and multiple band emission) will also be mentioned. PMID- 21629961 TI - Pyrroloquinoxaline hydrazones as fluorescent probes for amyloid fibrils. AB - Here we describe the identification and preliminary characterization of a new class of pyrrolo(imidazo)quinoxaline hydrazones as florescent probes for Abeta(1 42) fibrils. All the newly developed compounds were able to bind amyloid fibrils formed in vitro and some of them displayed an increase of their fluorescence upon binding. When tested on brain tissue preparations presenting Abeta deposits, the described hydrazones selectively stained amyloid structures and did not display aspecific binding. The hydrazones did not show antifibrillogenic activity and electron microscopy analysis revealed that they do not interfere with fibrils structure. The described pyrrolo(imidazo)quinoxalines could be useful for studying amyloid structures in vitro. Moreover, their experimentally proven ability to cross the blood-brain barrier in mouse opens the possibility of developing these compounds as potential amyloid imaging agents for in vivo applications. PMID- 21629962 TI - The coordination chemistry of Zn(II), Cd(II) and Hg(II) complexes with 1,2,4 triazole derivatives. AB - This perspective illustrates the coordination features of complexes constructed by 1,2,4-triazole derivatives and transition metal ions which belong to Group IIB, namely Zn(II), Cd(II) and Hg(II), demonstrates their behaviors in thermal stabilities, gas or liquid adsorption, fluorescence and nonlinear optical properties and also discusses the relation between their properties and crystal structures. Various 1,2,4-triazole derivatives containing versatile donor sites for coordination can be obtained through introducing different substituent groups to C3, N4 and C5 positions, thus offering rich coordination modes. The structures of these complexes rely on their triazole ligands, as well as mixed ligands, metal ions, anions and synthetic conditions. Obviously, the diversity in structure induces the controllability of properties, since the properties are influenced by several factors, which is significant for the applications of potential multifunctional materials. PMID- 21629963 TI - Ruthenium anticancer compounds: myths and realities of the emerging metal-based drugs. AB - Ruthenium anticancer drugs have attracted an increasing interest in the last 20 years and two of them have entered clinical trials. Compared to platinum drugs, the complexes based on ruthenium are often identified as less toxic and capable of overcoming the resistance induced by platinum drugs in cancer cells. These activities were attributed to the transportation to tumour cells by transferrin and to the selective activation to more reactive species by the reducing environment of solid tumours as compared to healthy tissues. Ruthenium anticancer drugs have been almost always designed to mimic platinum drugs, particularly for targeting DNA. Indeed, none of the above properties has never been clearly demonstrated even for the ruthenium drugs that entered clinical trials. The suggestion for the future is to change the perspective when designing new chemical entities, abandoning the philosophy that guided the actual panel of ruthenium drugs and to look further into the fine mechanism by which the most relevant ruthenium complexes available kill the target tumour cells, then focusing on targets selective of tumour cells and responsible for cell growth and malignancy. PMID- 21629964 TI - Definitive solid-state 185/187Re NMR spectral evidence for and analysis of the origin of high-order quadrupole-induced effects for I=5/2. AB - Rhenium-185/187 solid-state nuclear magnetic resonance (SSNMR) experiments using NaReO(4) and NH(4)ReO(4) powders provide unambiguous evidence for the existence of high-order quadrupole-induced effects (HOQIE) in SSNMR spectra. Fine structure, not predicted by second-order perturbation theory, has been observed in the (185/187)Re SSNMR spectrum of NaReO(4) at 11.75 T, where the ratio of the Larmor frequency (nu(0)) to the quadrupole frequency (nu(Q)) is ~2.6. This is the first experimental observation that under static conditions, HOQIE can directly manifest in SSNMR powder patterns as additional fine structure. Using NMR simulation software which includes the quadrupole interaction (QI) exactly, extremely large (185/187)Re nuclear quadrupole coupling constants (C(Q)) are accurately determined. QI parameters are confirmed independently using solid state (185/187)Re nuclear quadrupole resonance (NQR). We explain the spectral origin of the HOQIE and provide general guidelines that may be used to assess when HOQIE may impact the interpretation of the SSNMR powder pattern of any spin 5/2 nucleus in a large, axially symmetric electric field gradient (EFG). We also quantify the errors incurred when modeling SSNMR spectra for any spin-5/2 nucleus within an axial EFG using second-order perturbation theory. Lastly, we measure rhenium chemical shifts in the solid state for the first time. PMID- 21629965 TI - New insights on the mechanism of oxidation of D-galacturonic acid by hypervalent chromium. AB - The pollutant Cr(VI) is known to be very carcinogenic. In conditions of excess of Cr(VI), oxidation of D-galacturonic acid (Galur), the major metabolite of pectin, yields d-galactaric acid (Galar) and Cr(III). The redox reaction takes place through a multistep mechanism involving formation of intermediate Cr(II/IV) and Cr(V) species. The mechanism combines one- and two-electron pathways for the reduction of Cr(IV) by the organic substrate: Cr(VI)-> Cr(IV)-> Cr(II) and Cr(VI) > Cr(IV)-> Cr(III). This is supported by the observation of the optical absorption spectra of Cr(VI) esters, free radicals, CrO(2)(2+) (superoxoCr(III) ion) and oxo-Cr(V) complexes. Cr(IV) cannot be directly detected; however, formation of CrO(2)(2+) provides indirect evidence for the intermediacy of Cr(II/IV). Cr(IV) reacts with Galur much faster than Cr(V) and Cr(VI) do. The analysis of the reaction kinetics via optical absorption spectroscopy shows that the Cr(IV)-Galur reaction rate inversely depends on [H(+)]. Nevertheless, high [H(+)] still does not facilitate accumulation of Cr(IV) in the Cr(VI)-Galur mixture. Cr(VI) and the intermediate Cr(V) react with Galur at comparable rates; therefore the build-up and decay of Cr(V) accompany the decay of Cr(VI). The complete rate laws for the Cr(VI), Cr(V) and Cr(IV)-Galur redox reaction are here derived in detail. Furthermore, the nature of the five-co-ordinated oxo-Cr(V) bischelate complexes formed in Cr(VI)-Galur mixtures at pH 1-5 is investigated using continuous-wave and pulsed electron paramagnetic resonance (EPR) and density functional theory (DFT). PMID- 21629966 TI - Thrombin activatable fibrinolysis inhibitor. AB - Thrombin activatable fibrinolysis inhibitor (TAFI) was discovered two decades ago as a consequence of the identification of an unstable carboxypeptidase (CPU), which was formed upon thrombin activation of the respective pro-enzyme (proCPU). The antifibrinolytic function of the activated form (TAFIa, CPU) is directly linked to its capacity to remove C-terminal lysines from the surface of the fibrin clot. No endogenous inhibitors have been identified, but TAFIa activity is regulated by its intrinsic temperature-dependent instability with a half-life of 8 to 15 min at 37 degrees C. A variety of studies have demonstrated a role for TAFI/TAFIa in venous and arterial diseases. In addition, a role in inflammation and cell migration has been shown. Since an elevated level of TAFIa it is a potential risk factor for thrombotic disorders, many inhibitors, both at the level of activation or at the level of activity, have been developed and were proven to exhibit a profibrinolytic effect in animal models. Pharmacologically active inhibitors of the TAFI/TAFIa system may open new ways for the prevention of thrombotic diseases or for the establishment of adjunctive treatments during thrombolytic therapy. PMID- 21629967 TI - CALHM1 P86L polymorphism modulates CSF Abeta levels in cognitively healthy individuals at risk for Alzheimer's disease. AB - The calcium homeostasis modulator 1 (CALHM1) gene codes for a novel cerebral calcium channel controlling intracellular calcium homeostasis and amyloid-beta (Abeta) peptide metabolism, a key event in the etiology of Alzheimer's disease (AD). The P86L polymorphism in CALHM1 (rs2986017) initially was proposed to impair CALHM1 functionally and to lead to an increase in Abeta accumulation in vitro in cell lines. Recently, it was reported that CALHM1 P86L also may influence Abeta metabolism in vivo by increasing Abeta levels in human cerebrospinal fluid (CSF). Although the role of CALHM1 in AD risk remains uncertain, concordant data have now emerged showing that CALHM1 P86L is associated with an earlier age at onset of AD. Here, we have analyzed the association of CALHM1 P86L with CSF Abeta in samples from 203 AD cases and 46 young cognitively healthy individuals with a positive family history of AD. We failed to detect an association between the CALHM1 polymorphism and CSF Abeta levels in AD patients. Our data, however, revealed a significant association of CALHM1 P86L with elevated CSF Abeta42 and Abeta40 in the normal cohort at risk for AD. This work shows that CALHM1 modulates CSF Abeta levels in presymptomatic individuals, strengthening the notion that CALHM1 is involved in AD pathogenesis. These data further demonstrate the utility of endophenotype-based approaches focusing on CSF biomarkers for the identification or validation of risk factors for AD. PMID- 21629968 TI - Prognostic potential and tumor growth-inhibiting effect of plasma advanced glycation end products in non-small cell lung carcinoma. AB - The plasma fluorescence related to the standard fluorescence of advanced glycation end products (AGEs) is a simple measurable blood parameter for distinct diseases but its importance in human cancer, including non-small cell lung carcinoma (NSCLC), is unknown. Plasma samples of 70 NSCLC patients who underwent resection surgery of the tumor were analyzed for the distinct AGE-related fluorescence at 370 nm excitation/440 nm emission. In a retrospective study, we tested the prognostic relevance of this AGE-related plasma fluorescence. The effect of circulating AGEs on the NSCLC growth was studied experimentally in vitro and in vivo. NSCLC patients with high (> median) AGE-related plasma fluorescence were characterized by a later reoccurrence of the tumor after curative surgery and a higher survival rate compared with patients with low plasma fluorescence (25% versus 47% 5-y survival, P = 0.011). Treating NSCLC cell spheroids with patients' plasma showed an inverse correlation between the growth of spheroids in vitro and the individual AGE-related fluorescence of each plasma sample. To confirm the impact of circulating AGEs on the NSCLC progression, we studied the NSCLC growth in mice whose circulating AGE level was elevated by AGE rich diet. In vivo tumorigenicity assays demonstrated that mice with higher levels of circulating AGEs developed smaller tumors than mice with normal AGE levels. The AGE-related plasma fluorescence has prognostic relevance for NSCLC patients in whom the tumor growth-inhibiting effect of circulating AGEs might play a critical role. PMID- 21629970 TI - Dermatoscopy for Diagnosis of Creeping Hair: A Case Report and Mini-review of the Literature. PMID- 21629969 TI - Inhibition of c-Jun N-terminal kinase attenuates low shear stress-induced atherogenesis in apolipoprotein E-deficient mice. AB - Atherosclerosis begins as local inflammation of arterial walls at sites of disturbed flow, such as vessel curvatures and bifurcations with low shear stress. c-Jun NH2-terminal kinase (JNK) is a major regulator of flow-dependent gene expression in endothelial cells in atherosclerosis. However, little is known about the in vivo role of JNK in low shear stress in atherosclerosis. We aimed to observe the effect of JNK on low shear stress-induced atherogenesis in apolipoprotein E-deficient (ApoE(-/-)) mice and investigate the potential mechanism in human umbilical vein endothelial cells (HUVECs). We divided 84 male ApoE(-/-) mice into two groups for treatment with normal saline (NS) (n = 42) and JNK inhibitor SP600125 (JNK-I) (n = 42). Perivascular shear stress modifiers were placed around the right carotid arteries, and plaque formation was studied at low shear stress regions. The left carotid arteries without modifiers represented undisturbed shear stress as a control. The NS group showed atherosclerotic lesions in arterial regions with low shear stress, whereas the JNK-I group showed almost no atherosclerotic lesions. Corresponding to the expression of proatherogenic vascular cell adhesion molecule 1 (VCAM-1), phospho-JNK (p-JNK) level was higher in low shear stress regions with NS than with JNK-I inhibitor. In HUVECs under low shear stress, siRNA knockdown and SP600125 inhibition of JNK attenuated nuclear factor (NF)-kappaB activity and VCAM-1 expression. Furthermore, siRNA knockdown of platelet endothelial cell adhesion molecule 1 (PECAM-1) (CD31) reduced p-JNK and VCAM-1 levels after low shear stress stimulation. JNK may play a critical role in low shear stress-induced atherogenesis by a PECAM-1-dependent mechanosensory pathway and modulating NF kappaB activity and VCAM-1 expression. PMID- 21629971 TI - Evaluation of an IFN-gamma assay in the diagnosis of latent tuberculosis in patients with psoriasis in a highly endemic setting. AB - Screening for latent tuberculosis infection is mandatory before starting anti tumour necrosis factor treatments, but its diagnosis still poses a challenge. While studies performed in developed countries have demonstrated superior performance of T-cell based interferon-gamma release assay (IGRA) compared with the tuberculin skin test, there is a debate about whether this holds true in tuberculosis endemic areas. The performance of an IGRA kit T-SPOT.TB was evaluated in 33 moderate-to-severe untreated psoriasis patients and, as controls, 30 patients with common dermatological diseases at a tuberculosis highly endemic setting. The frequency of positive tuberculin skin test responses and induration size in controls were higher than in psoriasis patients (53% vs. 18% and 9.3 +/- 1.4 vs. 2.6 +/- 0.7 mm, respectively, p < 0.001). In contrast, the frequency of positive response and mean number of spots elicited with the T-SPOT.TB test were not significantly different between patients and controls (47% vs. 43% and 14.7 +/- 3.2 vs. 20.5 +/- 3.1 spots/well, respectively). The two tests presented good agreement in the control, but not the psoriasis group (kappa values of 0.625 and 0.375, respectively). Thus, in a highly tuberculosis-endemic setting the T SPOT.TB test was superior to the tuberculin skin test in diagnosing latent tuberculosis infection in psoriasis, probably because the immune dysregulation of psoriasis shows a lower interference in the in vitro test. PMID- 21629972 TI - Acute generalized exanthematous pustulosis due to etodolac in a patient with an iliopsoas muscle abscess. PMID- 21629973 TI - Severe systemic reaction to topical imiquimod. PMID- 21629974 TI - A dermoscopic figure of polymorphous atypical vessels with colour gradation. Metastatic cutaneous carcinoma into the lymphatic vessels. PMID- 21629975 TI - Systemic rituximab in multifocal primary cutaneous follicle centre lymphoma. AB - The aim of this retrospective study was to assess the efficacy and tolerance of intravenous rituximab in multifocal primary cutaneous follicle centre lymphomas (PCFCL). Eleven patients with a multifocal PCFCL were treated with rituximab (MabThera((r))) administrated intravenously. After four infusions, an objective response was observed in 90% of patients, and one month after all the infusions (median of 6 infusions) all the patients had an objective response and complete remission was obtained in 7 of 11 patients (64%). Follow-up ranged from 9 to 65 months (median: 30 months). Local disease recurrence was observed in five patients. The median progression-free survival time after the end of treatment was 23.6 months. This study is the largest series of patients with multifocal primary PCFCL treated with intravenous rituximab. This therapy is a safe and effective treatment and could represent an excellent alternative treatment to radiotherapy. PMID- 21629976 TI - Bullous dermolysis of the newborn and dystrophic epidermolysis bullosa pruriginosa within the same family: two phenotypes associated with a COL7A1 mutation. PMID- 21629977 TI - Lymphomatoid papulosis showing gammadelta T-cell phenotype. PMID- 21629978 TI - Papillary tumor of the pineal region. AB - Tumors of the pineal region are rare in adulthood, accounting for approximately 1% of intracranial neoplasms in this age range. Papillary tumor of the pineal region (PTPR) was first described by Jouvet et al. in 2003. In 2007, PTPR was included by the World Health Organization as a distinct entity in their new classification of central nervous system tumors. We report a case of PTPR, and undertake a literature review of this rare neuroepithelial neoplasm. Patients with PTPR need long-term follow up, and new cases should be well documented so that we can gain a better understanding of this neoplasm. PMID- 21629979 TI - Molecular and clinical analysis of glioblastoma with an oligodendroglial component (GBMO). AB - The genetic and clinical features of glioblastoma with an oligodendroglial component (GBMO), pathologically defined as anaplastic oligo-astrocytoma with necrosis, remain unclear. We investigated the correlation between genetic alterations and clinical outcomes in 19 GBMO patients we have encountered since 1997. Using single nucleotide polymorphism oligonucleotide genomic (SNP) microarrays, we analyzed gene amplification, loss of heterozygosity (LOH), and homozygous deletions in their whole genome. We also analyzed their overall survival (OS). Pathological studies revealed the presence of calcification in 11 and of a cyst in 9 of the 19 patients. Whole-genome analysis using SNP microarrays revealed LOH of chromosome 10 in 11, EGFR amplification in 8, 9p21 (INK4 locus) deletion in 12, PDGFR amplification in 2, and LOH of 1p19q in 2 patients. Median OS was 14 months (average 22.8 months). The pattern of genetic alterations was similar in GBMO and glioblastoma multiforme (GBM) patients, and the clinical outcomes were similar in GBMO and GBM patients. PMID- 21629980 TI - Loss of heterozygosity analysis in malignant gliomas. AB - Despite recent advances in the diagnosis and treatment of glioblastomas, patient outcomes for these highly malignant tumors remain poor. Research into the molecular pathology of glioblastoma has uncovered various genetic changes that contribute to malignancy. Some of the identified molecular markers--such as loss of heterozygosity (LOH) on chromosome 1p/19q and chromosome 10, O6-methylguanine methyltransferase promoter hypermethylation, and mutation of isocitrate dehydrogenase-1--may help to predict patient outcomes. Indeed, LOH analysis is an effective approach to classify malignant gliomas. Genome-wide analyses have revealed that the extent and pattern of LOH regions may have important implications for the clinical course of the disease. As the genetic underpinnings of malignant gliomas are complex and varied, careful selection of the methods for genetic analysis in the clinic is important. The fundamental principles of each assay need to be understood to allow careful selection of practically useful methods. This review summarizes recent developments in the molecular analysis of malignant glioma. PMID- 21629981 TI - A signal-inducing bone cement for magnetic resonance imaging-guided spinal surgery based on hydroxyapatite and polymethylmethacrylate. AB - The aim of this study was to develop a signal-inducing bone cement for magnetic resonance imaging (MRI)-guided cementoplasty of the spine. This MRI cement would allow precise and controlled injection of cement into pathologic lesions of the bone. We mixed conventional polymethylmethacrylate bone cement (PMMA; 5 ml methylmethacrylate and 12 g polymethylmethacrylate) with hydroxyapatite (HA) bone substitute (2-4 ml) and a gadolinium-based contrast agent (CA; 0-60 MUl). The contrast-to-noise ratio (CNR) of different CA doses was measured in an open 1.0 Tesla scanner for fast T1W Turbo-Spin-Echo (TSE) and T1W TSE pulse sequences to determine the highest signal. We simulated MRI-guided cementoplasty in cadaveric spines. Compressive strength of the cements was tested. The highest CNR was (1) 87.3 (SD 2.9) in fast T1W TSE for cements with 4 MUl CA/ml HA (4 ml) and (2) 60.8 (SD 2.4) in T1W TSE for cements with 1 MUl CA/ml HA (4 ml). MRI-guided cementoplasty in cadaveric spine was feasible. Compressive strength decreased with increasing amounts of HA from 46.7 MPa (2 ml HA) to 28.0 MPa (4 ml HA). An MRI-compatible cement based on PMMA, HA, and CA is feasible and clearly visible on MRI images. MRI-guided spinal cementoplasty using this cement would permit direct visualization of the cement, the pathologic process, and the anatomical surroundings. PMID- 21629982 TI - Case-based lung image categorization and retrieval for interstitial lung diseases: clinical workflows. AB - PURPOSE: Clinical workflows and user interfaces of image-based computer-aided diagnosis (CAD) for interstitial lung diseases in high-resolution computed tomography are introduced and discussed. METHODS: Three use cases are implemented to assist students, radiologists, and physicians in the diagnosis workup of interstitial lung diseases. RESULTS: In a first step, the proposed system shows a three-dimensional map of categorized lung tissue patterns with quantification of the diseases based on texture analysis of the lung parenchyma. Then, based on the proportions of abnormal and normal lung tissue as well as clinical data of the patients, retrieval of similar cases is enabled using a multimodal distance aggregating content-based image retrieval (CBIR) and text-based information search. The global system leads to a hybrid detection-CBIR-based CAD, where detection-based and CBIR-based CAD show to be complementary both on the user's side and on the algorithmic side. CONCLUSIONS: The proposed approach is in accordance with the classical workflow of clinicians searching for similar cases in textbooks and personal collections. The developed system enables objective and customizable inter-case similarity assessment, and the performance measures obtained with a leave-one-patient-out cross-validation (LOPO CV) are representative of a clinical usage of the system. PMID- 21629983 TI - Statistical shape and texture model of quadrature phase information for prostate segmentation. AB - PURPOSE: Prostate volume estimation from segmentation of transrectal ultrasound (TRUS) images aids in diagnosis and treatment of prostate hypertrophy and cancer. Computer-aided accurate and computationally efficient prostate segmentation in TRUS images is a challenging task, owing to low signal-to-noise ratio, speckle noise, calcifications, and heterogeneous intensity distribution in the prostate region. METHOD: A multi-resolution framework using texture features in a parametric deformable statistical model of shape and appearance was developed to segment the prostate. Local phase information of log-Gabor quadrature filter extracted texture of the prostate region in TRUS images. Large bandwidth of log Gabor filter ensures easy estimation of local orientations, and zero response for a constant signal provides invariance to gray level shift. This aids in enhanced representation of the underlying texture information of the prostate unaffected by speckle noise and imaging artifacts. The parametric model of the propagating contour is derived from principal component analysis of prior shape and texture information of the prostate from the training data. The parameters were modified using prior knowledge of the optimization space to achieve segmentation. RESULTS: The proposed method achieves a mean Dice similarity coefficient value of 0.95 +/- 0.02 and mean absolute distance of 1.26 +/- 0.51 millimeter when validated with 24 TRUS images of 6 data sets in a leave-one-patient-out validation framework. CONCLUSIONS: The proposed method for prostate TRUS image segmentation is computationally efficient and provides accurate prostate segmentations in the presence of intensity heterogeneities and imaging artifacts. PMID- 21629984 TI - Functional characterization of two new members of the caffeoyl CoA O methyltransferase-like gene family from Vanilla planifolia reveals a new class of plastid-localized O-methyltransferases. AB - Caffeoyl CoA O-methyltransferases (OMTs) have been characterized from numerous plant species and have been demonstrated to be involved in lignin biosynthesis. Higher plant species are known to have additional caffeoyl CoA OMT-like genes, which have not been well characterized. Here, we identified two new caffeoyl CoA OMT-like genes by screening a cDNA library from specialized hair cells of pods of the orchid Vanilla planifolia. Characterization of the corresponding two enzymes, designated Vp-OMT4 and Vp-OMT5, revealed that in vitro both enzymes preferred as a substrate the flavone tricetin, yet their sequences and phylogenetic relationships to other enzymes are distinct from each other. Quantitative analysis of gene expression indicated a dramatic tissue-specific expression pattern for Vp-OMT4, which was highly expressed in the hair cells of the developing pod, the likely location of vanillin biosynthesis. Although Vp-OMT4 had a lower activity with the proposed vanillin precursor, 3,4 dihydroxybenzaldehyde, than with tricetin, the tissue specificity of expression suggests it may be a candidate for an enzyme involved in vanillin biosynthesis. In contrast, the Vp-OMT5 gene was mainly expressed in leaf tissue and only marginally expressed in pod hair cells. Phylogenetic analysis suggests Vp-OMT5 evolved from a cyanobacterial enzyme and it clustered within a clade in which the sequences from eukaryotic species had predicted chloroplast transit peptides. Transient expression of a GFP-fusion in tobacco demonstrated that Vp-OMT5 was localized in the plastids. This is the first flavonoid OMT demonstrated to be targeted to the plastids. PMID- 21629986 TI - Depression among the Moroccan systemic sclerosis. AB - To determine the prevalence and predictors factors of depression in Moroccan patients with systemic sclerosis (SSc), we conducted a cross-sectional study of 59 Moroccan patients with systemic sclerosis. Patients were assessed by using the Patient Health Questionnaire depression scale "PHQ-9" and through extensive clinical histories and medical examinations. The Arabic version of HAQ and SF-36 was used to assess functional disability and health status, respectively. Forty six patients (77.4%) presented symptoms of depression. Thirty six (61%) have a major depressive syndrome and 10 (16.4%) have a minor depressive syndrome. The PHQ-9 score was significantly higher in the patient with prolonged disease duration, severe joint pain, higher disease severity, and important acute-phase reactants. Also, depression had a negative impact on physical and mental scores. Systemic scleroderma is associated with a high prevalence of depression. Screening for depression among patients with SSc is recommended and it should be assessed for routinely. PMID- 21629987 TI - Granulocyte colony stimulating factor in the treatment of cardiac ischemic disease. A decade has passed: is it time to give up? : editorial to "disassociation between left ventricular mechanical and electrical properties in ischemic rat heart after g-csf treatment" by Liu et al. PMID- 21629988 TI - Predictive and prognostic values of transient ischemic dilatation of left ventricular cavity for coronary artery disease and impact of various managements on clinical outcome using technetium-99m sestamibi gated myocardial perfusion imaging. AB - BACKGROUND: Transient ischemic dilatation (TID) of LV cavity during stress gated myocardial perfusion imaging (GMPI) is known as a predictor of severe CAD and signifies worse prognosis. OBJECTIVES: To assess predictive and prognostic value of TID of LV cavity using GMPI and clinical outcome in patients treated conservatively or with revascularization. METHODS: 189 patients out of 2689 were recruited (M:F 127/62, mean age 56 +/- 9 years) whose same-day stress GMPI revealed TID ratio (> 1.22) with no (sum stress score, SSS < 2) or reversible perfusion defects (sum difference score, SDS > 2). Coronary angiography (CA) was performed within 3 months in 125/189 cases who were followed for mean period of 18 +/- 4 months for fatal or non-fatal MI. RESULTS: CA was positive in 121/125 patients with TID for significant CAD (LAD = 11, multi vessel disease = 110 patients, positive predictive value 95%) and negative for obstructive disease in 4/125 (false-positive cases). 41/121 underwent revascularization within 2 months of CA (Intervention group), and remaining 80/121 were managed conservatively (Non Intervention group). Overall event rate was 20% (4/16%: fatal/non-fatal MIs). Kaplan-Meier survival curves revealed event-free survival in Intervention and Non Intervention groups for fatal MI 98/96% (P = 0.758), and for non-fatal MI, it was 97/58%, respectively (P = 0.042). CONCLUSION: We conclude that TID is a reliable predictor of multi vessel CAD and is associated with high incidence of non-fatal MIs than fatal MIs. Revascularization (PCI/CABG) rather than medical treatment should be considered in patients with TID for better clinical outcome. PMID- 21629989 TI - Spinal cord involvement in a child with raccoon roundworm (Baylisascaris procyonis) meningoencephalitis. AB - A 14-month-old previously healthy boy developed progressively worsening neurological symptoms secondary to eosinophilic meningoencephalitis with myelitis caused by raccoon roundworm (Baylisascaris procyonis) infection. MRI demonstrated T2 hyperintensity and enhancement of the cerebral white matter, cerebellum and spinal cord. Prior case reports have described signal abnormality within the brains of patients with raccoon roundworm neural larva migrans (NLM). This is a unique case in which spinal cord involvement was established by imaging. Knowledge of this combination of imaging findings expands the known imaging phenotype of this noteworthy infection. PMID- 21629990 TI - A phase II randomized study of cetuximab and bevacizumab alone or in combination with gemcitabine as first-line therapy for metastatic pancreatic adenocarcinoma. AB - The purpose of this study was to assess the efficacy and safety of bevacizumab plus cetuximab with or without gemcitabine in patients with advanced pancreatic adenocarcinoma. Patients with locally advanced or metastatic pancreatic adenocarcinoma, previously untreated, were randomized to bevacizumab (10 mg/kg q2w) plus cetuximab (400/250 mg/m(2) initial/weekly), either with (Arm A) or without (Arm B) gemcitabine (1000 mg/m(2) weekly * 3 of 4 weeks). Tumor assessments were performed q8w. Primary study endpoint was progression-free survival (PFS). Sixty-one patients were randomized to Arm A (n = 30) or Arm B (n = 31). Median treatment duration was 9 weeks in Arm A and 8 weeks in Arm B (range, 2.0-40.4). Patients in Arm A had median PFS and overall survival values of 3.55 months and 5.41 months, respectively, compared to 1.91 months and 4.17 months in Arm B. The study closed early due to lack of sufficient efficacy in both treatment arms. Although both regimens were well tolerated, patients treated with gemcitabine experienced more grade 3-4 toxicities, including proteinuria and thromboembolic events. The combination of cetuximab and bevacizumab did not result in promising activity with or without gemcitabine, suggesting that a strategy of dual EGFR/VEGF inhibition in pancreatic cancer does not warrant further development. To our knowledge, this is one of the first trials to evaluate a completely noncytotoxic regimen in the first-line treatment of advanced pancreatic cancer. (ClinicalTrials.gov number, NCT00326911). PMID- 21629991 TI - Severity of children's ADHD symptoms and parenting stress: a multiple mediation model of self-regulation. AB - The goal of the current study was to determine the extent to which the perceived self-regulation deficits across behavioral, cognitive, and emotional domains seen in children with ADHD explain the association between the severity of ADHD symptoms and parenting stress. Participants for this study included 80 children (mean age = 10 years, 9 months) with a DSM-IV diagnosis of ADHD confirmed by a comprehensive clinical diagnostic assessment. Parents reported their own stress levels as well as the severity of their children's ADHD symptoms, aggression, emotional lability, and executive functioning difficulties. Results indicated that the severity of children's hyperactivity/impulsivity symptoms but not their inattention related to parenting stress. Multiple mediational analyses indicated that the association between hyperactivity/impulsivity and parenting stress was explained by children's perceived comorbid aggression levels, emotional lability, and executive functioning difficulties. No significant differences in the strength of the mediators were found. The current study provides initial data showing that the perceived impairments in children's self-regulation across emotional, cognitive, and behavioral domains are what parents report as stressful, not simply the severity of ADHD symptoms. Due to the cross-sectional nature of this study and shared variance from relying solely on parent report, it will be critical for future research to replicate our findings using longitudinal and multi-informant data such as teacher reports and standardized assessments. PMID- 21629992 TI - Tamarind tree seed dispersal by ring-tailed lemurs. AB - In Madagascar, the gallery forests of the south are among the most endangered. Tamarind trees (Tamarindus indica) dominate these riverine forests and are a keystone food resource for ring-tailed lemurs (Lemur catta). At Berenty Reserve, the presence of tamarind trees is declining, and there is little recruitment of young trees. Because mature tamarinds inhibit growth under their crowns, seeds must be dispersed away from adult trees if tree recruitment is to occur. Ring tailed lemurs are likely seed dispersers; however, because they spend much of their feeding, siesta, and sleeping time in tamarinds, they may defecate a majority of the tamarind seeds under tamarind trees. To determine whether they disperse tamarind seeds away from overhanging tamarind tree crowns, we observed two troops for 10 days each, noted the locations of feeding and defecation, and collected seeds from feces and fruit for germination. We also collected additional data on tamarind seedling recruitment under natural conditions, in which seedling germination was abundant after extensive rain, including under the canopy. However, seedling survival to 1 year was lower when growing under mature tamarind tree crowns than when growing away from an overhanging crown. Despite low fruit abundance averaging two fruits/m(3) in tamarind crowns, lemurs fed on tamarind fruit for 32% of their feeding samples. Daily path lengths averaged 1,266 m, and lemurs deposited seeds throughout their ranges. Fifty-eight percent of the 417 recorded lemur defecations were on the ground away from overhanging tamarind tree crowns. Tamarind seeds collected from both fruit and feces germinated. Because lemurs deposited viable seeds on the ground away from overhanging mature tamarind tree crowns, we conclude that ring-tailed lemurs provide tamarind tree seed dispersal services. PMID- 21629993 TI - Prevention of hepatitis B virus reactivation in immunosuppressive therapy or chemotherapy. AB - In recent years, hepatitis B virus (HBV) has been found to reproliferate either during or following immunosuppressive therapy or chemotherapy, with hepatitis caused by HBV reactivation now considered a serious issue. HBV reactivation is categorized into occurrence in HBsAg- and anti-HBe-positive asymptomatic carriers, and in HBsAg-negative, anti-HBc low-titer-positive, and/or anti-HBs positive resolved HBV infection cases. Despite the fact that "clinical cure" is claimed for such resolved HBV cases, low levels of ongoing HBV production are now recognized as being sustained within the liver or in peripheral blood mononuclear cells, with the infection thus now considered to be virologically persistent. The risk of HBV reactivation rises as the level of immunosuppression intensifies, but in recent years HBV reactivation risk has been clearly shown to increase in cases of rituximab plus steroid-containing regimen for treatment of malignant lymphoma. In particular, the incidence of fulminant hepatitis caused by HBV reactivation in cases with resolved HBV infection is reported to be higher than that brought about by acute hepatitis B. Therefore, for all cases in which immunosuppressive therapy or chemotherapy treatment regimens are used, screening for HBV infection and appropriate management in accordance with the status of HBV-related markers are crucial, aimed at preventing occurrence of HBV reactivation. The foundation of the aforementioned management, regardless of HBsAg status, is administration of nucleoside analogues, with their powerful anti-viral properties, when HBV DNA levels reach detectable levels. PMID- 21629994 TI - Mechanism by which chronic kidney disease causes cardiovascular disease and the measures to manage this phenomenon. AB - In Japan, the number of chronic kidney disease (CKD) patients is thought to be 13,300,000, next in size after those with hypertension and diabetes. Although the number of patients with CKD seems large, it does not mean that all these patients require special treatment. Among them, nephrologists should pay special attention to patients with glomerular filtration rate below 50 mL/min/1.73 m(2) and progressive deterioration of renal function. Treatment of these CKD patients by a limited number of specialists is simply impossible; hence, it is essential to request treatment from physicians who are involved in general internal medicine and primary care. It is well known that not only does CKD cause end-stage renal failure, it also causes the onset of cardiovascular diseases (CVD) such as cardiac infarction and cerebral stroke; however, the question is how much significance does CKD have as a risk factor for CVD. It is understandable that hypertension and CVD are often complications of CKD; however, in addition to what is conventionally mentioned, there are three or four mechanisms that we would like to emphasize, and discuss herein. Among them, we would like to stress the role of klotho genes with special reference to the generation of CVD in CKD patients. When patients develop CKD, it is therefore necessary to remove as far as possible any factors that could represent a risk for CVD. Moreover, by taking appropriate measures against clinical conditions that often complicate CKD, such as hypertension, renal anemia, hyperuricemia, and hyperlipidaemia, the development of CVD can be prevented. PMID- 21629995 TI - Lung injury following acute kidney injury: kidney-lung crosstalk. AB - The mortality of acute kidney injury (AKI) remains unacceptably high, especially associated with acute respiratory failure. Lung injury complicated with AKI was previously considered as "uremic lung", which is characterized by volume overload and increased vascular permeability. New experimental data using rodent models of renal ischemia-reperfusion and bilateral nephrectomy have emerged recently focusing on kidney-lung crosstalk in AKI, and have highlighted the pathophysiological significance of increased cytokine concentration, enhanced inflammatory responses, and neutrophil activation. In this review, we outline the history of uremic lung and acute lung injury (ALI)/acute respiratory distress syndrome (ARDS), the epidemiological data on the synergistic effect of AKI and lung injury on mortality, and recent basic research which has identified possible pathways in AKI-induced lung injury. These findings will enable us to develop new therapeutic strategies against lung injury associated with AKI and improve the outcomes of critically ill patients in intensive care units. PMID- 21629996 TI - A role for neuropeptide Y Y5 but not the Y1-receptor subtype in food deprivation induced reinstatement of heroin seeking in the rat. AB - RATIONAL AND OBJECTIVES: Neuropeptide Y (NPY), an orexigenic peptide that is released during periods of food restriction, has been shown to have a significant modulatory impact on drug-related behaviors. We have previously reported that both acute food deprivation (FD) and NPY injections can reinstate extinguished drug-seeking behavior, a proposed animal model of relapse to drug abuse. However, it is not clear whether the FD effect on drug seeking is dependent on NPY transmission. Here, we used the reinstatement model to assess the role of NPY Y1 and Y5-receptor-mediated transmission in FD-induced reinstatement of heroin seeking. METHODS: Rats were trained to self-administer heroin for 10-12 days (0.1 mg/kg/infusion/intravenous). Animals then underwent extinction training followed by drug-seeking reinstatement tests under 21 h of FD and sated conditions. RESULTS: Injections of a novel NPY Y5-receptor antagonist, Lu AA33810 (0.0, 1.0, or 30.0 mg/kg/IP), resulted in a significant attenuation of FD-induced reinstatement of extinguished heroin seeking. However, no significant effects on reinstatement were found for the Y1-receptor antagonist, BIBO 3304 (0.0, 5.0, or 10.0 nmol/intracerebroventricular). CONCLUSIONS: These results suggest that while signals mediated through NPY Y1 receptors play a modest role in reinstatement, activation of Y5 receptors has a critical function in FD-induced reinstatement of heroin-seeking behavior. PMID- 21629997 TI - Prompt but inefficient: nicotine differentially modulates discrete components of attention. AB - RATIONALE: Nicotine has been shown to improve both memory and attention when assessed through speeded motor responses. Since very few studies have assessed effects of nicotine on visual attention using measures that are uncontaminated by motoric effects, nicotine's attentional effects may, at least partially, be due to speeding of motor function. OBJECTIVES: Using an unspeeded, accuracy-based test, the CombiTVA paradigm, we examined whether nicotine enhances attention when it is measured independently of motor processing. METHODS: We modelled data with a computational theory of visual attention (TVA; Bundesen 1990) so as to derive independent estimates of several distinct components of attention from performance of the single task: threshold of visual perception, perceptual processing speed, visual short-term memory storage capacity and top-down controlled selectivity. Acute effects of nicotine (2 mg gum) on performance were assessed in 24 healthy young non-smokers in a placebo-controlled counterbalanced, crossover design. Chronic effects of nicotine were investigated in 24 age- and education-matched minimally deprived smokers. RESULTS: Both an acute dose of nicotine in non-smokers and chronic nicotine use in temporarily abstaining smokers improved perceptual thresholds but slowed subsequent perceptual speed. Moreover, both acute and chronic nicotine use reduced attentional selectivity though visual short-term memory capacity was unimpaired. CONCLUSIONS: Nicotine differentially affected discrete components of visual attention, with acute and chronic doses revealing identical patterns of performance. We challenge prior reports of nicotine-induced speeding of information processing by showing, for the first time, that nicotine slows down perceptual processing speed when assessed using accuracy-based measures of cognitive performance. PMID- 21629998 TI - Management of complications after varicoportal anastomosis in liver transplantation. AB - The presence of portal vein thrombosis is a potential limitation for liver transplantation. An intraoperative diagnosis is linked to extensive surgical treatment and massive postoperative complications and mortality. We present a surgical less risky method for the treatment of intraoperatively diagnosed portal and mesenteric vein thrombosis that served as salvage therapy for a patient who underwent liver transplantation in our centre. Postoperative complications were ascites and renal failure. Persistent ascites required repeated paracentesis during the first month after liver transplantation but medical treatment sufficed thereafter. Moderate renal failure as defined by the K/DOQI-guidelines improved gradually and dialysis was never indicated. Six months after transplantation, the patient had normal liver function and adequate renal function. PMID- 21629999 TI - Treating primary insomnia: clinical effectiveness and predictors of outcomes on sleep, daytime function and health-related quality of life. AB - Primary insomnia, as defined by DSM-IV-TR, refers to a persistent sleep disturbance which is not connected to a current psychiatric or physical condition, but significantly impairs social and occupational functioning. This study explored the impact of Cognitive Behavioral Therapy for Insomnia (CBT-i) on sleep, daytime functioning and health-related quality of life (HRQoL). Next, we investigated which factors predicted positive treatment outcome by examining demographics, insomnia characteristics, baseline levels of daytime function, HRQoL, sleep-disruptive beliefs and psychological health on post-treatment sleep quality, daytime function and HRQoL. 138 consecutive primary insomnia patients completed questionnaires pre- and post-treatment and at 6 months follow-up. After CBT-i, robust clinical improvements were observed in sleep, daytime function and HRQoL, regardless of age, gender, type or duration of the complaint. Patients with pre-treatment severe insomnia, pronounced daytime impairment and low psychological well-being benefited most. PMID- 21630000 TI - Psychological distress in patients presenting for initial renal transplant evaluation. AB - The comorbidity of psychological disorders with end-stage renal disease (ESRD) presents challenges for renal transplantation, including increased likelihood of medication noncompliance and poorer quality of life. Estimates of rates and severity of affective and anxiety disorders have varied significantly across studies of renal transplant patients, possibly due in part to variation in the methodology and timing of evaluations. To this point, few researchers have examined the psychological condition of patients who are newly referred for renal transplantation. This study examined rates of psychological distress using the Patient Health Questionnaire (PHQ) in a sample of 518 ESRD patients at the specific time point of first contact with the transplant center. In this sample, 15.1% of patients endorsed symptoms consistent with a depressive condition and 7.6% of patients endorsed an anxiety condition. These rates were lower than expected, which may be due to decreased distress in this sample, selection biases, or underreporting of symptoms due to patients' motivation to present themselves positively. PMID- 21630001 TI - Human and economic burden of GAD, subthreshold GAD, and worry in a primary care sample. AB - Generalized anxiety disorder (GAD) has been associated with significant impairment and estimates of human and economic burden associated with the disorder are substantial. Little has been done, however, to examine impairment associated with subthreshold presentations of the disorder in medically underserved populations. This study compared primary care patients with GAD (n = 30), subthreshold GAD (n = 21), worry (n = 79), and no worry (n = 199) on measures of human and economic burden. On measures of human burden, all three symptomatic groups reported poorer perceived physical health, greater stress, and sleep difficulty. Worried and subthreshold groups also reported lower social support. For economic burden, GAD and worry groups reported a greater number of prescription medications. However, when co-morbid depression was accounted for the effect was no longer significant. Groups did not differ on employment status, number of visits to the clinic in the last 90 days, or physical health. Results are discussed in terms of identification, prevention, and intervention for GAD in primary care settings. PMID- 21630002 TI - Pain intensity, psychological inflexibility, and acceptance of pain as predictors of functioning in adolescents with juvenile idiopathic arthritis: a preliminary investigation. AB - Juvenile Idiopathic Arthritis (JIA) is a chronic rheumatic disease associated with pain and maladjustment. This study investigated whether pain, acceptance of pain, and psychological inflexibility uniquely predicted functional disability, anxiety, general quality of life (QOL), and health-related quality of life (HQOL) among adolescents with JIA. Twenty-three adolescents with JIA and pain were recruited from a pediatric rheumatology clinic. Participants completed self report measures pertaining to the key study variables. A series of multiple regression analyses demonstrated that higher pain uniquely predicted higher functional disability. Greater psychological inflexibility uniquely predicted higher anxiety, lower general QOL, and lower HQOL. Increases in acceptance of pain were found to be uniquely related to increases in general QOL. These data confirm prior findings that pain impacts functioning, and provide preliminary findings that psychological inflexibility and acceptance may be important targets of psychological intervention for youth with JIA and pain to improve functioning and QOL. PMID- 21630003 TI - Serum procalcitonin is not an early marker of pulmonary exacerbation in children with cystic fibrosis. AB - Serum procalcitonin (PCT) has been proposed as a marker to identify bacterial infection in children. For optimal management of cystic fibrosis (CF) patients, early recognition of pulmonary exacerbations is necessary, but sensitive biomarkers to do so are lacking. Our study was done to establish baseline values for PCT in children with CF and to compare these to values at onset of a pulmonary exacerbation. Serum PCT values were determined in CF children during an outpatient clinic visit and at onset of treatment with intravenous (IV) antibiotics for a pulmonary exacerbation. Serum PCT was measured using a quantitative immunoassay (BRAHMS Kryptor PCTsensitive, Henningsdorf, Germany). In 92 outpatients (mean age 10.0 years, SD 4.8 years; mean forced expiratory volume in 1 s 91%, SD 18; 9 chronically colonized with Pseudomonas aeruginosa), mean baseline PCT was 0.05 ng/ml (SD 0.07). Mean PCT on admission for IV treatment of pulmonary exacerbation was 0.07 ng/ml (SD 0.06) (n = 22) and not different from the baseline value. PCT values were markedly higher in two CF patients with an acute nonrespiratory infection (central venous catheter-associated bloodstream infection, acute gastroenteritis), demonstrating that they can mount a PCT response. CONCLUSION: PCT values in CF children are not different from values reported in healthy children. In CF children, PCT values do not rise significantly at the onset of a respiratory exacerbation and thus hold no promise as an early marker to identify a pulmonary exacerbation. PMID- 21630005 TI - Are water vole resistant to anticoagulant rodenticides following field treatments? AB - The anti-vitamin Ks (AVKs) are widely used to control rodent populations. They inhibit Vitamin K regeneration by the Vitamin K Epoxide Reductase (VKOR) and cause a fatal hemorrhagic syndrome. Because of repeated use, some populations of commensal rodents have expressed resistance to these compounds. In Franche-Comte (France), the water vole exhibits cyclic population outbreaks. A second generation AVK, bromadiolone, has been used for the last 20 years to control vole populations. The aim of this study is to determine whether these repeated treatments could have led to the development of resistance to AVKs in water vole populations. We conducted enzymatic and genetic studies on water voles trapped in treated and non treated plot. The results indicate that voles from the most heavily treated area exhibit enzymatic changes in VKOR activity hence arguing for resistance to AVKs and that an intronic haplotype on the vkorc1 gene seems to be associated with these enzymatic changes. PMID- 21630006 TI - IT delivery of ChABC modulates NG2 and promotes GAP-43 axonal regrowth after spinal cord injury. AB - Chondroitin sulphate proteoglycans (CSPGs) with the major component NG2 have an inhibitory effect on regeneration of damaged axons after spinal cord injury. In this study, we investigate whether the digestion of CSPGs by chondroitinase ABC (ChABC) may decrease the NG2 expression and promote axon regrowth through the lesion site. Rats underwent spinal cord compression injury and were treated with ChABC or vehicle through an intrathecal catheter delivery at 2, 3, and 4 days after injury. In addition, animals were behaviorally scored using BBB test in weekly intervals after SCI. Based on immunocytochemical analyses, we have quantified distribution of NG2 glycoprotein and GAP-43 in spinal cord tissue in both experimental groups. Multiple injections of ChABC caused decrease of NG2 expression at lesion site at 5 and 7 days, but not at 14 and 28 days in comparison with vehicle-treated rats and significantly enhanced GAP-43 expression during the entire survival. The densitometry analysis showed significantly higher GAP-43 immunoreactivity (1.8-2.2-fold) in the regrowing axons and cell bodies within the central lesion cavity when compared with vehicle group. Longitudinally oriented and disorganized GAP-43-labeled axons were able to infiltrate and penetrate damaged tissue. The outgrowth of GAP-43 axons after CHABC delivery was significantly longer (<=0.457 mm) when compared with the length of axons in vehicle-treated rats (<=0.046 mm). Present findings suggest that degradation of NG2 with acute IT ChABC treatment may promote ongoing (long-lasting) axonal regenerative processes at late survival (14 and 28 days), but with no significant impact on the improvement of motor function. PMID- 21630007 TI - Transplantation of predifferentiated adipose-derived stromal cells for the treatment of spinal cord injury. AB - Adipose-derived stromal cells (ASCs) are an alternative source of stem cells for cell-based therapies of neurological disorders such as spinal cord injury (SCI). In the present study, we predifferentiated ASCs (pASCs) and compared their behavior with naive ASCs in vitro and after transplantation into rats with a balloon-induced compression lesion. ASCs were predifferentiated into spheres before transplantation, then pASCs or ASCs were injected intraspinally 1 week after SCI. The cells' fate and the rats' functional outcome were assessed using behavioral, histological, and electrophysiological methods. Immunohistological analysis of pASCs in vitro revealed the expression of NCAM, NG2, S100, and p75. Quantitative RT-PCR at different intervals after neural induction showed the up regulated expression of the glial markers NG2 and p75 and the neural precursor markers NCAM and Nestin. Patch clamp analysis of pASCs revealed three different types of membrane currents; however, none were fast activating Na(+) currents indicating a mature neuronal phenotype. Significant improvement in both the pASC and ASC transplanted groups was observed in the BBB motor test. In vivo, pASCs survived better than ASCs did and interacted closely with the host tissue, wrapping host axons and oligodendrocytes. Some transplanted cells were NG2- or CD31-positive, but no neuronal markers were detected. The predifferentiation of ASCs plays a beneficial role in SCI repair by promoting the protection of denuded axons; however, functional improvements were comparable in both the groups, indicating that repair was induced mainly through paracrine mechanisms. PMID- 21630008 TI - Special Issue on A Tribute to Marshall Nirenberg. Introduction. PMID- 21630009 TI - Marshall Nirenberg 1927-2010. PMID- 21630010 TI - Activation of liver X receptor decreases BACE1 expression and activity by reducing membrane cholesterol levels. AB - The synthetic Liver X receptor (LXR) activator T0901317 has been reported to exert neuroprotective effect in Alzheimer's disease, but the relationship between LXR activation and beta-site amyloid precursor protein cleaving enzyme 1 (BACE-1) remains uncertain. This study investigated the effect of T0901317 on membrane cholesterol levels, BACE1 expression and activity. We found that T0901317 decreased membrane cholesterol levels, reduced BACE1 expression and activity as well as beta-secretase cleaved C-terminal fragment (beta-CTF) levels in vivo and in vitro. Meanwhile, the expression of ATP-binding membrane cassette transport protein A1 (ABCA1) enhanced. Additionally, inhibition of ABCA1 abrogated the effects of T0901317 on membrane cholesterol levels and beta-secretase activity. Moreover, addition of LXR antagonist reversed the effect of T0901317 on ABCA1 mRNA expression, membrane cholesterol levels and beta-secretase activity. Our results suggest that activation of LXR may decrease BACE1 expression and activity through a pathway associated with ABCA1-mediated reduction in membrane cholesterol levels. PMID- 21630011 TI - Correlates of HIV testing uptake among kothi-identified men who have sex with men in public sex environments in Chennai, India. AB - Kothi-identified men who have sex with men in India are highly marginalized and are at high-risk for HIV. This study examines HIV testing among 132 self-reported HIV-negative and unknown serostatus kothis recruited from public sex environments in Chennai, India. Using logistic regression we identified variables associated with HIV testing uptake (i.e., being tested and knowing the result). Sixty-one percent reported HIV testing uptake. At the bivariate level, married men, those with low HIV transmission knowledge, those who engaged in unprotected anal sex and unprotected receptive anal sex were at lower odds of reporting testing uptake. In multivariate analysis, married men and those with low levels of HIV transmission knowledge were at decreased odds of being tested, as were kothis who experienced forced sex. Culturally competent programs engaging married kothis are needed. Interventions to facilitate HIV prevention education and systemic interventions to combat sexual violence may facilitate HIV testing uptake among kothis. PMID- 21630012 TI - The critical role of intimacy in the sexual risk behaviors of gay and bisexual men. AB - Research indicates that high numbers of gay and bisexual men report infrequent or inconsistent condom use, placing them at risk for HIV and other STDs. The present study examined positive and negative condom-related attitudes along three dimensions-risk reduction, pleasure reduction, and intimacy interference-and examined their relative predictive power in determining condom use among a sample of sexually risky gay and bisexual men in New York City. In a multivariate model, both risk reduction and intimacy interference attitudes emerged as significant predictors of unprotected sex; however, the variance accounted for by a model including intimacy interference was almost three times that accounted for by a model including risk reduction alone. These data suggest a pivotal role for intimacy in shaping condom attitudes and behavior among gay and bisexual men. HIV prevention interventions should consider incorporating intimacy as a motivating factor for sexual behavior and a potential barrier to condom use. PMID- 21630014 TI - Homophobia and racism experienced by Latino men who have sex with men in the United States: correlates of exposure and associations with HIV risk behaviors. AB - Using cross-sectional data collected from 1081 Latino men who have sex with men (MSM) recruited with respondent-driven sampling (RDS) techniques from Los Angeles and New York, we examined the extent to which Latino MSM reported exposure to social discrimination (i.e., experienced both homophobia and racism, homophobia only, racism only, or neither homophobia nor racism). More than 40% of respondents experienced both homophobia and racism in the past 12 months. Los Angeles participants, those with lower income, and those who reported being HIV positive were more likely to report experiencing both types of social discrimination. Adjusting for potential confounders, men exposed to both homophobia and racism were more likely than men exposed to neither form of discrimination to report unprotected receptive anal intercourse with a casual sex partner (AOR = 1.92, 95% CI, 1.18-3.24) and binge drinking (AOR = 1.42, 95% CI, 1.02-1.98). Our findings suggest the presence of a syndemic of adverse social experiences and call for more intervention research to address both homophobia and racism experienced among Latino MSM in the United States. PMID- 21630013 TI - Outcomes of antiretroviral therapy in the Swiss HIV Cohort Study: latent class analysis. AB - An in-depth understanding of the different groups that make up the HIV-infected population should inform prevention and care. Using latent class analysis (LCA) we identified seven groups with similar socio-demographic and behavioral characteristics at enrolment in the Swiss HIV Cohort Study: older gay men, younger gay men, older heterosexual men, injection drug users, single migrants, migrant women in partnerships and heterosexual men and women. Outcomes of combination antiretroviral therapy (ART) were analyzed in 1,633 patients starting ART. Compared to older gay men, the probability of a virologic response to ART was reduced in single migrants, in older heterosexual men and in IDUs. Loss to follow-up was higher in single migrants and IDUs, and mortality was increased in older heterosexual men and IDUs. Socio-behavioral groups identified by LCA allow insights above what can be gleaned from traditional transmission groups, and may identify patients who could benefit from targeted interventions. PMID- 21630015 TI - Sex and relationships on the street: how homeless men judge partner risk on Skid Row. AB - Homeless men in the U.S. represent a large and growing population, and have elevated rates of HIV/AIDS and sexual risk behaviors, including unprotected sex with women. We conducted qualitative interviews (n = 30) with homeless men using shelters and meal lines in downtown Los Angeles (Skid Row) to better understand how such men view the risks of sexual encounters with female partners. Men living on Skid Row perceived multiple risks, including HIV and unwanted pregnancy as well as emotional trauma, loss of resources, exacerbation of drug addiction, and physical attack. Respondents described using visual and behavioral cues, social reputation, geographical location, feelings of trust, perceived relationship seriousness, and medically inaccurate "folk" beliefs to judge whether partners were risky and/or condom use was warranted. Medically inaccurate beliefs suggest the potential utility of evidence-based interventions to change such beliefs. We also consider implications for relationships on the street and housing interventions. PMID- 21630016 TI - Defective nuclear translocation of stress-activated signaling in senescent diploid human fibroblasts: a possible explanation for aging-associated apoptosis resistance. AB - In order to study the nature of aging-dependent apoptosis resistance, we compared the activation pattern of mitogen-activated protein kinases (MAPK) in response to three different stress modalities: hydrogen peroxide (H(2)O(2)), staurosporine, and thapsigargin. We observed the agonist-specific activation pattern of MAP kinases in human diploid fibroblasts (HDFs). When young HDFs were treated with PD98059, a specific inhibitor of extracellular signal-regulated kinase (ERK), H(2)O(2)-induced apoptosis was blocked, whereas staurosporine-induced apoptosis was inhibited by treatment with SB203580, a specific inhibitor of p38. In addition, the levels of anti-apoptotic protein Bcl-2 (B-cell lymphoma protein-2) were restored by PD98059 or SB239063 in cells treated with H(2)O(2) or staurosporine, respectively. We also found that inhibition of the nuclear import of p-Erk and p-p38 using wheat germ agglutinin induced apoptosis resistance in young HDF cells in response to H(2)O(2) or staurosporine. These data indicate a potential role of the nuclear translocation of apoptotic signals in the induction of apoptosis. Moreover, the nuclear translocation of activated ERK1/2 and p38 in response to H(2)O(2) or staurosporine was significantly compromised in senescent HDFs, compared with young cells. Taken together, we propose that the apoptosis resistance of senescent HDFs might be related to the defective nuclear translocation of stress-activated signals in an agonist-specific manner, which would imply the operation of an aging-dependent functional nucleo-cytoplasmic trafficking barrier. PMID- 21630017 TI - Pro-apoptotic cell death genes, hid and reaper, from the tephritid pest species, Anastrepha suspensa. AB - Pro-apoptotic proteins from the reaper, hid, grim (RHG) family are primary regulators of programmed cell death in Drosophila due to their antagonistic effect on inhibitor of apoptosis (IAP) proteins, thereby releasing IAP-inhibition of caspases that effect apoptosis. Using a degenerate PCR approach to conserved domains from the 12 Drosophila species, we have identified the first reaper and hid orthologs from a tephritid, the Caribfly Anastrepha suspensa. As-hid is the first identified non-drosophilid homolog of hid, and As-rpr is the second non drosophilid rpr homolog. Both genes share more than 50% amino acid sequence identity with their Drosophila homologs, suggesting that insect pro-apoptotic peptides may be more conserved than previously anticipated. Importantly, both genes encode the conserved IBM and GH3 motifs that are key for IAP-inhibition and mitochondrial localization. Functional verification of both genes as cell death effectors was demonstrated by cell death assays in A. suspensa embryonic cell culture, as well as in heterologous Drosophila melanogaster S2 cells. Notably, heterologous cell death activity was found to be higher for Anastrepha genes than their Drosophila counterparts. In common with the Drosophila cognates, As-hid and As-rpr negatively regulated the Drosophila inhibitor of apoptosis (DIAP1) gene to promote apoptosis, and both genes when used together effected increased cell death activity, indicating a co-operative function for As-hid and As-rpr. We show that these tephritid cell death genes are functional and potent as cell death effectors, and could be used to design improved transgenic lethality systems for insect population control. PMID- 21630018 TI - Propofol increases the rate of albumin-unbound free midazolam in serum albumin solution. AB - Propofol and midazolam have a synergistic anesthetic action. One of the reasons for this is thought to be the inhibitory effect of propofol on midazolam metabolism. However, because both drugs bind strongly to serum protein, their interaction may not only involve the effects of propofol on midazolam metabolism, but may also involve propofol's effects on serum protein-binding. Against this background, we investigated the characteristics of midazolam binding to serum albumin, and evaluated the effects of both propofol and ketamine on this binding. Midazolam was added to a serum albumin solution with propofol or ketamine, and, after incubation for 1 h, albumin-free solution was separated from the sample and the midazolam concentration was measured using a high-performance liquid chromatography system. The albumin-unbound rate of midazolam was evaluated and compared with the rate in the control solution (only midazolam). Propofol significantly raised the rate of albumin-unbound free midazolam, while ketamine had no effect on the binding of midazolam to serum albumin. These findings suggest that the increase in albumin-unbound free midazolam brought about by propofol is involved in the synergistic effect of these two agents. PMID- 21630019 TI - Feasibility and results of a modified intrafollicular insemination technique for treating primary infertility. PMID- 21630020 TI - Psychometric properties of the Spence Children's Anxiety Scale (SCAS) in Cypriot children and adolescents. AB - The Spence Children's Anxiety Scale (SCAS) is a 38-item self-report questionnaire which measures symptoms of DSM-IV anxiety disorders in children and adolescents. This study examined the psychometric properties of the Greek translation of SCAS in a large community sample of children and adolescents (N = 1,072), aged 12-17 years, in the non-occupied territory of Cyprus. A subsample of these participants was retested 8 weeks after the initial assessment. The SCAS demonstrated high internal consistency (alpha = .92) and test-retest reliability (r = .88). The SCAS total scores correlated significantly with the anxious/depressed subscale of the Youth Self-Report, the Columbia Impairment Scale, and with the emotional subscale of the Strength and Difficulties Questionnaire. Confirmatory factor analyses revealed the same six-factor structure as the original SCAS. The SCAS proved to be a reliable and valid measure of anxiety symptoms in the Cypriot context. PMID- 21630021 TI - Manipulation of saponin biosynthesis by RNA interference-mediated silencing of beta-amyrin synthase gene expression in soybean. AB - Soybean seeds contain substantial amount of diverse triterpenoid saponins that influence the seed quality, although little is known about the physiologic functions of saponins in plants. We now describe the modification of saponin biosynthesis by RNA interference (RNAi)-mediated gene silencing targeted to beta amyrin synthase, a key enzyme in the synthesis of a common aglycon of soybean saponins. We identified two putative beta-amyrin synthase genes in soybean that manifested distinct expression patterns with regard to developmental stage and tissue specificity. Given that one of these genes, GmBAS1, was expressed at a much higher level than the other (GmBAS2) in various tissues including the developing seeds, we constructed two RNAi vectors that encode self-complementary hairpin RNAs corresponding to the distinct regions of GmBAS1 under the control of a seed-specific promoter derived from the soybean gene for the alpha' subunit of the seed storage protein beta-conglycinin. These vectors were introduced independently into soybean. Six independent transgenic lines exhibited a stable reduction in seed saponin content, with the extent of saponin deficiency correlating with the beta-amyrin synthase mRNA depletion. Although some transgenic lines produced seeds almost devoid of saponins, no abnormality in their growth was apparent and the antioxidant activity of their seeds was similar to that of control seeds. These results suggest that saponins are not required for seed development and survival, and that soybean seeds may therefore be amenable to the modification of triterpenoid saponin content and composition through molecular biologic approaches. PMID- 21630022 TI - Synthesis and field tests of possible minor components of the sex pheromone of Prionus californicus. AB - Earlier work has shown that adult male Prionus californicus Motschulsky (Coleoptera: Cerambycidae) are attracted to the female-produced compound (3R,5S) 3,5-dimethyldodecanoic acid, and to a synthetic mixture of the four stereoisomers of 3,5-dimethyldodecanoic acid. Here, we report the results of field trials that tested whether or not three structurally related compounds (methyl 3,5 dimethyldodecanoate, 3,5-dimethyltridecanoic acid, and 3,5-dimethylpentadecanoic acid), present in extracts of virgin females, are attractive, and whether or not they influence attraction to 3,5-dimethyldodecanoic acid. In a trial with single components, only traps baited with the acid or its methyl ester captured more beetles than did control traps; catches to the acid were five times higher than to the methyl ester. Another trial, excluding 3,5-dimethyldodecanoic acid, confirmed the activity of the methyl ester. Finally, addition of the three compounds to 3,5-dimethyldodecanoic acid, in the ratio found in extracts from female beetles, gave a catch similar to that of traps baited with 3,5 dimethyldodecanoic acid alone. Consequently, the function of these minor compounds remains undetermined. PMID- 21630023 TI - A novel approach using telomerase-specific replication-selective adenovirus for detection of circulating tumor cells in breast cancer patients. AB - The aim of this study was to develop a new method for detecting circulating tumor cells (CTCs) in breast cancer patients by using the telomerase-specific replication-selective adenovirus OBP-401. Once transfected, OBP-401 can replicate only in telomerase expressing cells and emit fluorescence as it replicates so that the transfected cells become easily recognizable. Peripheral blood samples were drawn from 50 metastatic breast cancer patients and 27 early breast cancer patients. Blood samples were subjected to both the OBP-401 and CellSearch assays for the detection of CTCs and the results were compared. The recovery rate of the OBP-401 assay was one CTC in 7.5 ml blood combined with high specificity since no CTC was observed in 80 healthy controls. In 50 metastatic patients, 21 patients (42%) were identified as positive with the OBP-401 assay and 27 patients (54%) with the CellSearch assay. The CellSearch assay showed a significantly higher positivity for hormone receptor (HR)-positive tumors (estrogen receptor and/or progesterone receptor-positive tumors) (61%, 25/41, P = 0.012) or CA15-3-positive tumors (69%, 24/35, P = 0.003) than for HR-negative tumors (13%, 1/8) or CA15-3 negative tumors (21%, 3/14), respectively. Contrary, the OBP-401 assay results were similar regardless of their HR status (positive: 44% vs. negative: 38%, P = 0.738) or CA15-3 positivity (positive: 40% vs. negative: 50%, P = 0.523). Of the 27 early stage patients, four patients (15%) were identified by the OBP-401 assay and by the CellSearch assay, respectively, but there was no overlap in the CTCs positive patients. In conclusion, the OBP-401 assay is comparable to the CellSearch assay in the detection rate of CTCs in both metastatic and early breast cancer patients. However, there was a great discrepancy in patients with CTCs between both assays. The OBP-401 assay may isolate CTCs with other biological characteristics which CTCs detected by the CellSearch assay do not have. PMID- 21630024 TI - Single nucleotide polymorphisms in the 20q13 amplicon genes in relation to breast cancer risk and clinical outcome. AB - The 20q13 region is frequently amplified/overexpressed in breast tumours. However, the nature of this amplification/overexpression is unknown. Here, we investigated genetic variation in five 20q13 amplicon genes (MYBL2, AURKA, ZNF217, STK4 and PTPN1) and its impact on breast cancer (BC) susceptibility and clinical outcome. As a novel finding, four polymorphisms in STK4 (rs6017452, rs7271519) and AURKA (rs2273535, rs8173) associated with steroid hormone receptor status both in a Swedish population-based cohort of 783 BC cases and in a Polish familial/early onset cohort of 506 BC cases. In the joint analysis, the minor allele carriers of rs6017452 had more often hormone receptor positive tumours (OR 0.57, 95% CI 0.40-0.81), while homozygotes for the minor allele of rs7271519, rs2273535 and rs8173 had more often hormone receptor negative tumours (2.26, 1.30 3.39; 2.39, 1.14-5.01; 2.39, 1.19-4.80, respectively) than homozygotes for the common allele. BC-specific survival analysis of AURKA suggested that the Swedish carriers of the minor allele of rs16979877, rs2273535 and rs8173 might have a worse survival compared with the major homozygotes. The survival probabilities associated with the AURKA genotypes depended on the tumour phenotype. In the Swedish case-control study, associations with BC susceptibility were observed in a dominant model for three MYBL2 promoter polymorphisms (rs619289, P = 0.02; rs826943, P = 0.03 and rs826944, P = 0.02), two AURKA promoter polymorphisms (rs6064389, P = 0.04 and rs16979877, P = 0.02) and one 3'UTR polymorphism in ZNF217 (rs1056948, P = 0.01). In conclusion, our data confirmed the impact of the previously identified susceptibility locus and provided preliminary evidence for novel susceptibility variants in BC. We provided evidence for the first time that genetic variants at 20q13 may affect hormone receptor status in breast tumours and influence tumour aggressiveness and survival of the patients. Future studies are needed to confirm the prognostic value of our findings in the clinic. PMID- 21630025 TI - Evidence for sustained ATP release from liver cells that is not mediated by vesicular exocytosis. AB - Extracellular ATP regulates many important cellular functions in the liver by stimulating purinergic receptors. Recent studies have shown that rapid exocytosis of ATP-enriched vesicles contributes to ATP release from liver cells. However, this rapid ATP release is transient, and ceases in ~30 s after the exposure to hypotonic solution. The purpose of these studies was to assess the role of vesicular exocytosis in sustained ATP release. An exposure to hypotonic solution evoked sustained ATP release that persisted for more than 15 min after the exposure. Using FM1-43 (N-(3-triethylammoniumpropyl)-4-(4 (dibutylamino)styryl)pyridinium dibromide) fluorescence to measure exocytosis, we found that hypotonic solution stimulated a transient increase in FM1-43 fluorescence that lasted ~2 min. Notably, the rate of FM1-43 fluorescence and the magnitude of ATP release were not correlated, indicating that vesicular exocytosis may not mediate sustained ATP release from liver cells. Interestingly, mefloquine potently inhibited sustained ATP release, but did not inhibit an increase in FM1-43 fluorescence evoked by hypotonic solution. Consistent with these findings, when exocytosis of ATP-enriched vesicles was specifically stimulated by 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB), mefloquine failed to inhibit ATP release evoked by NPPB. Thus, mefloquine can pharmacologically dissociate sustained ATP release and vesicular exocytosis. These results suggest that a distinct mefloquine-sensitive membrane ATP transport may contribute to sustained ATP release from liver cells. This novel mechanism of membrane ATP transport may play an important role in the regulation of purinergic signaling in liver cells. PMID- 21630026 TI - The major lipid cores of the archaeon Ignisphaera aggregans: implications for the phylogeny and biosynthesis of glycerol monoalkyl glycerol tetraether isoprenoid lipids. AB - The lipid cores from Ignisphaera aggregans, a hyperthermophilic Crenarchaeon recently isolated from New Zealand hot springs, have been profiled by liquid chromatography-tandem mass spectrometry. The distribution revealed includes relatively high proportions of monoalkyl (also known as H-shaped) tetraether cores which have previously been implicated as kingdom-specific biomarkers for the Euryarchaeota. Such high expression of monoalkyl tetraether lipids is unusual in the archaeal domain and may indicate that formation of these components is an adaptive mechanism that allows I. aggregans to regulate membrane behaviour at high temperatures. The observed dialkyl tetraether and monoalkyl tetraether lipid distributions are similar but not fully concordant, showing differences in the average number of incorporated rings. The similarity supports a biosynthetic route to the ring-containing dialkyl and monoalkyl tetraether lipids via a dialkyl tetraether core containing zero rings, or a closely related structural relative, as an intermediate. Currently, however, the precise nature of the biosynthetic route to these lipids cannot be deduced. PMID- 21630028 TI - Amelioration of chronic pelvic sepsis secondary to a non-collapsible pelvic cavity using a gracilis rotational flap. PMID- 21630027 TI - Regulation of expression of trehalose-6-phosphate synthase during cold shock in Arthrobacter strain A3. AB - Trehalose is a chemical chaperone known to protect a variety of organisms against cold stress. Members of the genus Arthrobacter, which belongs to the Actinomycetales group, exhibit strong resistance to stress conditions, but exactly how trehalose synthesis is regulated in conditions of cold stress is still unknown. Here, we report that Arthrobacter strain A3, which was isolated from the alpine permafrost, has only two trehalose synthesis pathways (OtsA/B and TreS), while other Arthrobacter spp. have three. Mutants and immunoblot analyses indicate that trehalose is mainly synthesized via OtsA at low temperatures in Arthrobacter strain A3. Therefore, we have focused on the regulation of OtsA expression during cold shock. The results indicated that both low temperature and accumulation of trehalose can inhibit OtsA expression. The elongation factor Tu, which binds to OtsA, stabilizes the expression of OtsA in the cold. PMID- 21630029 TI - Abstracts of the Irish Society for Rheumatology and Irish Rheumatology Health Professionals Society Autumn Scientific Meeting. September 23-24, 2010. Dublin, Ireland. PMID- 21630030 TI - Frequency of the SLCO1B1 388A>G and the 521T>C polymorphism in Tanzania genotyped by a new LightCycler(r)-based method. AB - PURPOSE: The 388A>G and the 521T>C polymorphism of the SLCO1B1 gene affect the activity of the uptake transporter OATP1B1, thus influencing kinetics, safety, and efficacy of substrate drugs. To evaluate the impact of these polymorphisms in populations of different ethnic origins, it is important to know their frequencies and to develop fast and reliable methods for genotyping. We therefore established a new, high-throughput method and determined the genotype and allelic frequencies of these polymorphisms in Tanzanians, for which the frequencies were unknown thus far. METHODS: New LightCycler(r) 480-based methods with hybridization probes were established and used to genotype 366 Tanzanian and 236 European individuals for 388A>G (rs2306283) and 521T>C (rs4149056) in the SLCO1B1 gene. The methods were validated by direct sequencing of the polymerase chain reaction (PCR) products of heterozygous individuals and checked for intrarun precision repeatability, interrun precision reproducibility, robustness, and deviations from the Hardy-Weinberg equilibrium. RESULTS: The new methods allow unambiguous identification of the corresponding genotypes. There was a clear difference in allelic distribution between Tanzanians and Europeans, with the 388A>G (rs2306283) being much more prevalent in Tanzanians (87% vs 41%) and the 521T>C (rs4149056) being very rare in this African population (6% vs 17%). CONCLUSIONS: We developed robust and high-throughput methods to genotype common SLCO1B1 allelic variants with the LightCycler(r) 480. In Tanzanians, we identified the highest frequency of the 388A>G and 521T>C polymorphisms ever reported from black populations. The clinical relevance of SLCO1B1 genetic variation in the African population remains to be investigated. PMID- 21630031 TI - BCHE and CYP2D6 genetic variation in Alzheimer's disease patients treated with cholinesterase inhibitors. AB - PURPOSE: Cholinesterase inhibitors are commonly prescribed to patients with Alzheimer's disease (AD) to enhance cholinergic neurotransmission. Differential response to these treatments has been observed, and claims have been made that individual genetic variants may influence the pharmacokinetic and pharmacodynamic properties of these drugs. Here we assess the effects of genetic variation at two loci involved in the activity of cholinesterase inhibitors on longitudinal clinical change in AD patients being treated with donepezil, galantamine, and rivastigmine. METHODS: This was an open study in which 171 Italian AD patients treated with donepezil (n = 92), galantamine (n = 33), or rivastigmine (n = 46) were enrolled. Response to treatment was quantified by grading the patient's cognitive state (Mini-Mental State Examination) and the patient's ability to perform normal daily activities (Activities of Daily Living, Instrumental Activities of Daily Living) at baseline and after 6 and 12 months of treatment. Genetic variation was comprehensively characterized and analyzed at two loci: CYP2D6, which is involved in donepezil and galantamine metabolism, and BCHE, which codes for an enzyme (butyrylcholinesterase) which is both target and metabolizer of rivastigmine. APOE (coding for apolipoprotein E), which is associated with the risk of AD and inefficacy of specific AD treatments, was genotyped to control for patient stratification. The influence of the CYP2D6 and BCHE genotype on clinical changes after 12 months was evaluated by several tests of association. RESULTS: After 1 year of treatment, 29, 12, and 12 of the patients receiving donepezil, galantamine, and rivastigmine, respectively, showed a cognitive decrement, while eight patients interrupted the therapy before 12 months of treatment. No significant differences between the three treatments were observed in terms of response and tolerability. Non-responders show a higher proportion of BCHE and CYP2D6 mutated alleles, but genetic variation at the two loci was not a reliable predictor of clinical changes in AD patients treated with cholinesterase inhibitors. CONCLUSIONS: Individualized therapy based on CYP2D6 and BCHE genotypes is unlikely to be beneficial for treating Alzheimer's disease patients in routine clinical practice. PMID- 21630032 TI - The effect of bezafibrate and omega-3 fatty acids on lymphocyte cytokine release and systemic inflammation in patients with isolated hypertriglyceridemia. AB - PURPOSE: The aim of this study was to compare the effects of fibrates and omega-3 fatty acids on lymphocyte secretory function and systemic inflammation in patients with isolated hypertriglyceridemia. METHODS: The study included 107 patients with isolated hypertriglyceridemia who received bezafibrate (200 mg twice daily), omega-3 fatty acids (1 g twice daily) or placebo for 12 weeks. The lipid profile, fasting and 2-h post-glucose load plasma glucose levels, homeostasis model assessment index (HOMA), plasma high-sensitivity C-reactive protein (hsCRP) levels and lymphocyte release of interleukin-2, interferon-gamma and tumor necrosis factor-alpha were assessed at baseline, on the day of randomization, and after 4 and 12 weeks of treatment. RESULTS: Both bezafibrate and omega-3 fatty acids reduced plasma triglyceride levels. Bezafibrate additionally decreased total and low-density lipoprotein-cholesterol levels and the HOMA and insignificantly decreased post-glucose load plasma glucose, as well as increased high-density lipoprotein-cholesterol. Bezafibrate treatment was associated with a reduction in lymphocyte release of interleukin-2, interferon gamma and tumor necrosis factor-alpha, which was accompanied by a reduction in plasma hsCRP levels. Omega-3 fatty acid did not significantly reduce lymphocyte cytokine release and plasma hsCRP. The anti-inflammatory effects of both drugs did not correlate with their action on plasma lipids, but in the case of the former the effect was related to the improvement in insulin sensitivity. CONCLUSION: Our results indicate that bezafibrate is superior to omega-3 fatty acid in inhibiting systemic inflammation and lymphocyte secretory function. PMID- 21630033 TI - Call for participation in the neurogenetics consortium within the Human Variome Project. AB - The rate of DNA variation discovery has accelerated the need to collate, store and interpret the data in a standardised coherent way and is becoming a critical step in maximising the impact of discovery on the understanding and treatment of human disease. This particularly applies to the field of neurology as neurological function is impaired in many human disorders. Furthermore, the field of neurogenetics has been proven to show remarkably complex genotype-to-phenotype relationships. To facilitate the collection of DNA sequence variation pertaining to neurogenetic disorders, we have initiated the "Neurogenetics Consortium" under the umbrella of the Human Variome Project. The Consortium's founding group consisted of basic researchers, clinicians, informaticians and database creators. This report outlines the strategic aims established at the preliminary meetings of the Neurogenetics Consortium and calls for the involvement of the wider neurogenetic community in enabling the development of this important resource. PMID- 21630034 TI - Non-verbal memory measured by Rey-Osterrieth Complex Figure B: normative data. AB - One of the major problems that clinical neuropsychology has had in memory clinics is to apply ecological, easily administrable and sensitive tests that can make the diagnosis of dementia both precocious and reliable. Often the choice of the best neuropsychological test is hard because of a number of variables that can influence a subject's performance. In this regard, tests originally devised to investigate cognitive functions in healthy adults are not often appropriate to analyze cognitive performance in old subjects with low education because of their intrinsically complex nature. In the present paper, we present normative values for the Rey-Osterrieth Complex Figure B Test (ROCF-B) a simple test that explores constructional praxis and visuospatial memory. We collected normative data of copy, immediate and delayed recall of the ROCF-B in a group of 346 normal Italian subjects above 40 years. A multiple regression analysis was performed to evaluate the potential effect of age, sex, and education on the three tasks administered to the subjects. Age and education had a significant effect on copying, immediate recall, and delayed recall as well as on the rate of forgetting. Correction grids and equivalent scores with cut-off values relative to each task are available. The availability of normative values can make the ROCF-B a valid instrument to assess non-verbal memory in adults and in the elderly for whom the commonly used ROCF-A is too demanding. PMID- 21630035 TI - Pituitary tumor apoplexy presenting as infective meningoencephalitis. AB - We report on a case of a 80-year-old man who developed progressive drowsiness with headache, fever and signs of meningeal irritation 2 days after a head trauma. Suspecting an infective meningoencephalitis, the patient was treated with wide spectrum antibiotic and antiviral therapy. Brain CT scan revealed a previously unknown pituitary expansive lesion. A brain MRI study confirmed the presence of an intrasellar lesion, which presented remarkable contrast ring enhancement, and showed non-specific inflammatory tissue on the clivus, possibly responsible of the clinical features of sterile meningitis. A biopsy proven diagnosis of pituitary apoplexy was made. This case highlights MRI as an important investigation for earlier recognition of pituitary apoplexy that can present with a clinical picture resembling an infective meningoencephalitis. PMID- 21630036 TI - The concept of FDG-PET endophenotype in Alzheimer's disease. AB - Often viewed as a potential tool for preclinical diagnosis in early asymptomatic stages of Alzheimer's disease (AD), the term "endophenotype" has acquired a recent popularity in the field. In this review, we analyze the construct of endophenotype-originally designed to discover genes, and examine the literature on potential endophenotypes for the late-onset form of AD (LOAD). We focus on the [18F]-fluoro-2-deoxyglucose (FDG) PET technique, which shows a characteristic pattern of hypometabolism in AD-related regions in asymptomatic carriers of the ApoE E4 allele and in children of AD mothers. We discuss the pathophysiological significance and the positive predictive accuracy of an FDG-endophenotype for LOAD in asymptomatic subjects, and discuss several applications of this endophenotype in the identification of both promoting and protective factors. Finally, we suggest that the term "endophenotype" should be reserved to the study of risk factors, and not to the preclinical diagnosis of LOAD. PMID- 21630037 TI - Epilepsy in coeliac disease: not just a matter of calcifications. AB - The clinical spectrum of epilepsy related to celiac disease (CD) ranges from benign syndromes to intractable epilepsy with evolution to a severe encephalopathy, including progressive myoclonic epilepsy (PME). A more specific syndrome characterised by the association of CD, epilepsy, and occipital calcifications (CEC) has also been reported. This study describes the clinical, neuroradiological and neurophysiological features of eight consecutive epileptic patients with a diagnosis of CD confirmed by laboratory tests and duodenal biopsy, referring to our Epilepsy Centre. Despite its small size, this series reflects the broad spectrum of the association between the two diseases, since it includes four cases of CEC and a more heterogeneous group of patients without cerebral calcifications comprising one case of limbic encephalitis and a case of PME. Our cohort suggests that more complex pathogenic mechanisms may be involved in the association between epilepsy and CD, and that CD should be included in the screening for PME etiology. Our data also confirm the major involvement of the occipital lobe, and minimise both the importance of calcifications in epileptogenesis and folic acid deficit in the development of calcifications. PMID- 21630038 TI - Neuropsychological evaluation and follow up in jcv- and non-jcv-related leukoencephalopathies in HIV infection. AB - The introduction of highly active antiretroviral therapy does not seem to have altered the incidence of progressive multifocal leukoencephalopathy (PML) in HIV infection. Moreover, the occurrence of a HIV-related leukoencephalopathy, called not determined leukoencephalopaties (NDLE), has been reported. As neuropsychological impairment remains highly prevalent in HIV infection, the aim of this study is to describe the neuropsychological profile of PML and NDLE patients, analyzing the time-related changes. Clinical and neuropsychological data from 32 patients (17 PML, 15 NDLE) were compared with two control groups: (1) asymptomatic HIV+ patients without magnetic resonance imaging evidence of leukoencephalopathy; (2) age-/gender-/education-matched healthy subjects. Patients with rapidly worsening PML were significantly impaired on all neuropsychological tests, while PML with more benign course and NDLE groups showed a dysexecutive pattern of impairment. Asymptomatic HIV+ subjects showed mild and isolated cognitive deficits, without functional impact. Cognitive impairment should therefore be considered a key feature from HIV infection diagnosis. PMID- 21630039 TI - Tetraethylammonium enhances the rectal and colonic motility in rats and human in vitro. AB - Hirschsprung's disease is the congenital absence of generating the peristaltic contractions transmitting from the proximal colon to rectum. We previously have found that tetraethylammonium (TEA), the nonselective Ca(2+)-activated K(+) channel blocker, increases the maximal contractile force and the amplitude of the contraction in rat duodenum. The present study is to test the effect of TEA on motility of colon and rectum from rats and Hirschsprung's disease patients in vitro, in order to find an alternative method to improve the syndrome of Hirschsprung's disease. The rectal and colonic motility was recorded by a tension transducer connected to a biology function experiment system. Histology was analyzed with standard hematoxylin and eosin staining. TEA (1, 3, and 5 mM) significantly increased the amplitude and frequency of contractility of colon and rectum from rats in longitudinal and circular direction. TEA at 5 and 15 mM concentrations showed no effect on histology of colon and rectum from rats that were administered locally with TEA into colon lumen from anus for 10 days. TEA at 15 mM increased the amplitude and frequency of contractions of the colon and rectum from Hirschsprung's disease patients. Our data showed that TEA increased the contractility of colon and rectum from rats and Hirschsprung's disease patients in vitro, suggesting that local administration of TEA in colon or rectum lumen might be an alternative method to ameliorate the syndrome of Hirschsprung's disease patients who are not cured completely by surgery or not suitable for surgery. PMID- 21630040 TI - Ontogeny of salinity tolerance and evidence for seawater-entry preparation in juvenile green sturgeon, Acipenser medirostris. AB - We measured the ontogeny of salinity tolerance and the preparatory hypo osmoregulatory physiological changes for seawater entry in green sturgeon (Acipenser medirostris), an anadromous species occurring along the Pacific Coast of North America. Salinity tolerance was measured every 2 weeks starting in 40 day post-hatch (dph) juveniles and was repeated until 100% survival at 340/00 was achieved. Fish were subjected to step increases in salinity (50/00 12 h(-1)) that culminated in a 72-h exposure to a target salinity, and treatment groups (0, 15, 20, 25, 30, 340/00; and abrupt exposure to 340/00) were adjusted as fish developed. After 100% survival was achieved (134 dph), a second experiment tested two sizes of fish for 28-day seawater (330/00) tolerance, and gill and gastrointestinal tract tissues were sampled. Their salinity tolerance increased and plasma osmolality decreased with increasing size and age, and electron microscopy revealed three types of mitochondria-rich cells: one in fresh water and two in seawater. In addition, fish held on a natural photoperiod in fresh water at 19 degrees C showed peaks in cortisol, thyroid hormones and gill and pyloric ceca Na(+), K(+)-ATPase activities at body sizes associated with seawater tolerance. Therefore, salinity tolerance in green sturgeon increases during ontogeny (e.g., as these juveniles may move down estuaries to the ocean) with increases in body size. Also, physiological and morphological changes associated with seawater readiness increased in freshwater-reared juveniles and peaked at their seawater-tolerant ages and body sizes. Their seawater-ready body size also matched that described for swimming performance decreases, presumably associated with downstream movements. Therefore, juvenile green sturgeon develop structures and physiological changes appropriate for seawater entry while growing in fresh water, indicating that hypo-osmoregulatory changes may proceed by multiple routes in sturgeons. PMID- 21630041 TI - Automated motion estimation of root responses to sucrose in two Arabidopsis thaliana genotypes using confocal microscopy. AB - Root growth is a highly dynamic process influenced by genetic background and environment. This paper reports the development of R scripts that enable root growth kinematic analysis that complements a new motion analysis tool: PlantVis. Root growth of Arabidopsis thaliana expressing a plasma membrane targeted GFP (C24 and Columbia 35S:LTI6b-EGFP) was imaged using time-lapse confocal laser scanning microscopy. Displacement of individual pixels in the time-lapse sequences was estimated automatically by PlantVis, producing dense motion vector fields. R scripts were developed to extract kinematic growth parameters and report displacement to +/- 0.1 pixel. In contrast to other currently available tools, Plantvis-R delivered root velocity profiles without interpolation or averaging across the root surface and also estimated the uncertainty associated with tracking each pixel. The PlantVis-R analysis tool has a range of potential applications in root physiology and gene expression studies, including linking motion to specific cell boundaries and analysis of curvature. The potential for quantifying genotype * environment interactions was examined by applying PlantVis R in a kinematic analysis of root growth of C24 and Columbia, under contrasting carbon supply. Large genotype-dependent effects of sucrose were recorded. C24 exhibited negligible differences in elongation zone length and elongation rate but doubled the density of lateral roots in the presence of sucrose. Columbia, in contrast, increased its elongation zone length and doubled its elongation rate and the density of lateral roots. PMID- 21630042 TI - Structure and regulation of the Asr gene family in banana. AB - Abscisic acid, stress, ripening proteins (ASR) are a family of plant-specific small hydrophilic proteins. Studies in various plant species have highlighted their role in increased resistance to abiotic stress, including drought, but their specific function remains unknown. As a first step toward their potential use in crop improvement, we investigated the structure and regulation of the Asr gene family in Musa species (bananas and plantains). We determined that the Musa Asr gene family contained at least four members, all of which exhibited the typical two exons, one intron structure of Asr genes and the "ABA/WDS" (abscisic acid/water deficit stress) domain characteristic of Asr genes. Phylogenetic analyses determined that the Musa Asr genes were closely related to each other, probably as the product of recent duplication events. For two of the four members, two versions corresponding to the two sub-genomes of Musa, acuminata and balbisiana were identified. Gene expression and protein analyses were performed and Asr expression could be detected in meristem cultures, root, pseudostem, leaf and cormus. In meristem cultures, mAsr1 and mAsr3 were induced by osmotic stress and wounding, while mAsr3 and mAsr4 were induced by exposure to ABA. mASR3 exhibited the most variation both in terms of amino acid sequence and expression pattern, making it the most promising candidate for further functional study and use in crop improvement. PMID- 21630043 TI - Palliative functional hemispherectomy for treatment of refractory status epilepticus associated with Alpers' disease. AB - PURPOSE: Palliative epilepsy surgery is considered for patients that would benefit from surgical therapy for intractable epilepsy but are not candidates for curative procedures. In many cases, the goals of therapy focus on improved quality of life more than seizure freedom. We discuss the use of epilepsy surgery for refractory status epilepticus, as well as the rationale and ethical considerations for employing a palliative procedure in otherwise fatal diseases. METHODS: We present a child with Alpers' disease presenting with refractory status epilepticus which was treated with functional hemispherectomy after failure of multiple typical therapies. Hemispherectomy allowed for the child to be extubated and ultimately discharged to home with her family. Unfortunately, the child died several months later after developing new-onset liver failure in the setting of a viral illness. CONCLUSION: Functional hemispherectomy was effective for the treatment of refractory status epilepticus in Alpers' disease. We believe that the procedure resulted in improved quality of life which was the primary outcome goal. Palliative procedures should be considered in diseases with ultimate fatal outcome when the short-term benefits outweigh the risks. The ethical aspects of treatment must be carefully considered to insure treatment is provided in the best interest of the patient. PMID- 21630044 TI - Fusion of a family 9 cellulose-binding module improves catalytic potential of Clostridium thermocellum cellodextrin phosphorylase on insoluble cellulose. AB - Clostridium thermocellum cellodextrin phosphorylase (CtCDP), a single-module protein without an apparent carbohydrate-binding module, has reported activities on soluble cellodextrin with a degree of polymerization (DP) from two to five. In this study, CtCDP was first discovered to have weak activities on weakly water soluble celloheptaose and insoluble regenerated amorphous cellulose (RAC). To enhance its activity on solid cellulosic materials, four cellulose binding modules, e.g., CBM3 (type A) from C. thermocellum CbhA, CBM4-2 (type B) from Rhodothermus marinus Xyn10A, CBM6 (type B) from Cellvibrio mixtus Cel5B, and CBM9 2 (type C) from Thermotoga maritima Xyn10A, were fused to the C terminus of CtCDP. Fusion of any selected CBM with CtCDP did not influence its kinetic parameters on cellobiose but affected the binding and catalytic properties on celloheptaose and RAC differently. Among them, addition of CBM9 to CtCDP resulted in a 2.7-fold increase of catalytic efficiency for degrading celloheptaose. CtCDP CBM9 exhibited enhanced specific activities over 20% on the short-chain RAC (DP = 14) and more than 50% on the long-chain RAC (DP = 164). The chimeric protein CtCDP-CBM9 would be the first step to construct a cellulose phosphorylase for in vitro hydrogen production from cellulose by synthetic pathway biotransformation (SyPaB). PMID- 21630045 TI - Uptake of KRAS mutation testing in patients with metastatic colorectal cancer in Europe, Latin America and Asia. AB - The mutation status of the KRAS gene in the tumors of patients with metastatic colorectal cancer (mCRC) is a predictive biomarker for the efficacy of epidermal growth factor receptor monoclonal antibody therapy. The establishment of KRAS mutation testing in this setting represents a significant change to standard diagnostic procedures and a major advance in the personalization of cancer care. Against a changing regulatory background, three cross-sectional surveys of physicians in 14 countries in Europe, Latin America and Asia were conducted in 2008, 2009, and 2010 to investigate the uptake and outcome of KRAS testing for patients with mCRC. Physicians in each year answered questions on four patients (last patient seen and last seen in first-, second- and third-line settings). Fieldwork was carried out February-May 2008, January-April 2009, and January April 2010. Data from 3,819, 3,740 and 3,820 anonymized, uncoded patient records were collated. The frequency of KRAS testing in patients with mCRC increased from 3% in 2008 to 47% in 2009 and 69% in 2010. The 2010 survey revealed that test results were available within 15 days for 82%, 51% and 98% of the 1679, 679, and 261 tested patients in the European, Latin American and Asian regions, respectively. Cetuximab was the most commonly administered targeted agent in tested patients with KRAS wild-type mCRC (798/1607 patients; 50%) and bevacizumab was the most commonly administered targeted agent in tested patients with KRAS mutant tumors (396/893; 44% overall). In conclusion, KRAS testing is now widely established as a routine diagnostic procedure for patients with mCRC and is used increasingly to guide treatment selection. PMID- 21630046 TI - Late cardiotoxicity after low dose of anthracycline therapy for acute lymphoblastic leukemia in childhood. AB - INTRODUCTION: Late cardiotoxicity is a known complication of anthracycline therapy but the long-term effects of low cumulative doses are not well documented. We studied late cardiotoxicity in survivors of childhood acute lymphoblastic leukemia (ALL) treated with low anthracycline doses 10 to 20 years earlier. METHODS: Seventy-seven ALL survivors who received a cumulative anthracycline dose <250 mg/m2 and were at least 10 years after treatment were evaluated for signs of clinical heart failure. Cardiac function was assessed by echocardiography including tissue Doppler measurements of the septal mitral annulus in 37 ALL survivors 10.6-18.3 years (median 13.3 years) after anthracycline treatment with cumulative doses of 180 (n = 19) or 240 mg/m2 (n = 18). The control group consisted of 30 healthy volunteers matched for age, sex, BSA, and BMI. RESULTS: No clinical relevant cardiotoxicity was found. Left ventricular shortening fraction (SF) was significantly reduced in male ALL survivors. Three of the 19 male ALL survivors had an SF below 30%. Male ALL survivors showed a significantly lower early filling velocity to atrial contraction velocity ratio but myocardial velocity during early filling was comparable between patients and controls. ALL survivors had a significantly longer isovolumetric relaxation time (IVRT). Thirty percent of the ALL survivors have an abnormal IVRT compared to the normal range of the controls. CONCLUSION AND IMPLICATIONS FOR CANCER SURVIVORS: At a median of 13.3 years after exposure to cumulative doses of anthracyclines of 180 or 240 mg/m2, no clinical relevant cardiotoxicity was found but subclinical cardiac abnormalities were present in 30% of the patients. PMID- 21630047 TI - A randomised controlled trial for the effectiveness of intra-articular Ropivacaine and Bupivacaine on pain after knee arthroscopy: the DUPRA (DUtch Pain Relief after Arthroscopy)-trial. AB - PURPOSE: In this double-blinded, randomised clinical trial, the aim was to compare the analgesic effects of low doses of intra-articular Bupivacaine and Ropivacaine against placebo after knee arthroscopy performed under general anaesthesia. METHODS: A total of 282 patients were randomised to 10 cc NaCl 0.9%, 10 cc Bupivacaine 0.5% or 10 cc Ropivacaine 0.75%. Patients received the assigned therapy by intra-articular injection after closure of the portal. Pain and satisfaction were measured at one, 4 h and 5-7 days after arthroscopy with Numerical Rating Scale (NRS) -scores. NSAID consumption was also recorded. RESULTS: One-h NRS-scores at rest were higher in the NaCl group compared with the Bupivacaine group (P < 0.01), 1 h NRS-scores in flexion were higher in the NaCl group compared with the Bupivacaine (P < 0.01) and Ropivacaine (P < 0.01) groups. NRS-satisfaction at 4 h was higher for the Bupivacaine group compared with the NaCl group (P = 0.01). Differences in NRS-scores were significant but low in magnitude. NSAID consumption was lower in the Bupivacaine group compared with the NaCl group (P < 0.01). CONCLUSIONS: The results of this randomised clinical trial demonstrate improved analgesia after administration of low doses of intra articular Bupivacaine and Ropivacaine after arthroscopy of the knee. Considering reports of Bupivacaine and Ropivacaine being chondrotoxic agents and the relatively small improvement on patient comfort found in this trial, it is advised to use systemic anaesthetic instead of intra-articular Bupivacaine or Ropivacaine for pain relief after knee arthroscopy. PMID- 21630048 TI - No difference in anterior tibial translation with and without posterior cruciate ligament in less invasive total knee replacement. AB - PURPOSE: The relative advantages of cruciate retaining or cruciate resecting total knee replacement are still controversial. If the posterior cruciate ligament (PCL) is preserved, it should be properly balanced. In a previous study, it was demonstrated that increasing the flexion gap leads to an anterior translation of the tibia relative to the femur. Based on these results, we hypothesized that cutting the PCL increases the flexion gap and lessens anterior tibial translation. METHODS: The amount of anterior tibial translation versus distraction force in the flexion gap was measured in 88 total knee replacements with a less invasive midvastus approach using a custom-made tensioner. Measurements were performed with intact and resected PCL. RESULTS: The difference in tibial translation with and without PCL is not significant. A 1-mm increase in the flexion gap led to an average anterior translation of 0.6 mm with intact PCL and 0.4 mm with cut PCL, which is less than that reported in a previous study. CONCLUSIONS: The results have not confirmed our initial hypothesis. The reasons for this may be other soft tissue structures that prevent anterior tibial translation, such as the collateral ligaments, and/or the extensor apparatus. Moreover, the knee flexion angle for the used specific implant may play a role. LEVEL OF EVIDENCE: Prospective comparative study, Level II. PMID- 21630049 TI - Education in arthroscopy, sports medicine and knee surgery. PMID- 21630050 TI - Improved stability with intramedullary stem after anterior femoral notching in total knee arthroplasty. AB - PURPOSE: It has been hypothesized that femoral notching in total knee arthroplasties weakens the cortex of the femur, which can predispose to femoral fractures in the postoperative period. Some authors suggest that patients who sustain inadvertent notching should have additional protection in the postoperative period, and consideration should be given to the use of prophylactic femoral stems. In this case, a question can be raised: Is the use of femoral stem in an anterior femoral notching an effective way to reduce the fracture risk? We hypothesized that for a larger notch, the use of a femoral stem does not decrease considerably the stress-riser at the notch edge, and the use of stem is not enough to reduce the risk of fracture. METHODS: In the present in vitro study, twelve synthetic femurs were selected and used for the experiments under two load scenarios. Femoral components with and without femoral stems were implanted in femurs with different notch sizes to predict experimentally the strain levels at the notch edge with the use of fiber Bragg gratings and at notch region with strain gauges. RESULTS: Despite the global strain reduction in stemmed condition, at the notch edge, the strain behavior was dissimilar for the different notch depths. For notch depths lower than 5 mm, the use of stem reduces the strain level at the notch edge to values below the intact femur condition, while for depths greater or equal to 5 mm, the strain levels at the notch edge were higher than the intact femur condition with values ranging from +10 to +189%. CONCLUSIONS: The present study suggests the use of a prophylactic stem for notch depths greater than 5 mm. For notch depths below 5 mm, the fracture risk due to strain increase at the notch edge seems to be low in the stemless condition. PMID- 21630051 TI - Use of pineapple juice with gadopentetate dimeglumine as a negative oral contrast for magnetic resonance cholangiopancreatography: a multicentric study. AB - We evaluated the efficacy of pineapple juice with gadopentetate dimeglumine as a negative oral contrast agent for magnetic resonance cholangiopancreatography (MRCP). Images were obtained before and after the intake of a negative oral contrast agent. Images obtained from six different areas of the biliary tree were analyzed by three different radiologists, who were blind to the exams; scores regarding image quality were given to each area. The statistical analysis showed a significant difference between images before and after the use of the contrast agent (P < 0.001) for the three radiologists (R1-R3). Mean scores given by radiologists before the intake of the contrast agent were 2.49 +/- 0.42 (R1), 2.62 +/- 0.32 (R2), and 2.22 +/- 0.46 (R3). After the intake, mean scores were 3.38 +/- 0.62 (R1), 3.48 +/- 0.55 (R2), and 2.89 +/- 0.69 (R3). The ducts that showed the highest scores were the common bile duct and duct of Wirsung, the distal portion of the common bile duct and the cystic duct. We suggest herein that the contrast agent pineapple juice with gadopentate dimeglumine constitutes an efficient negative oral contrast agent for MRCP, for it efficiently eliminates the signal of the digestive tube in MRCP images. PMID- 21630052 TI - Screening of the common culture conditions affecting crystallinity of bacterial cellulose. AB - By analyzing with X-ray diffraction and FT-IR spectroscopy, the main effective factors among common culture conditions on crystallinity index and I(alpha) fraction of cellulose produced by Gluconacetobacter xylinus were examined with Plackett-Burman design experiment. Varying pH value in the medium by adjusting the composition of citrate buffer or by adding HCl/NaOH solution indicates it is the content of citrate buffer rather than its function of pH buffering that gives the influence on crystallinity. Further experiment reveals that Na+ concentration of 0.174 mol/l in medium with citrate buffer added would decrease the crystallinity index significantly. Comparison of carbon sources shows that fructose leads to a higher crystallinity index than glucose, which suggests a relationship between crystallinity and production speed of bacterial cellulose affected by carbon sources. An interesting phenomenon was that a longer period of cultivation would decrease the crystallinity of bacterial cellulose. The reason is assumed to be the dense network of cellulose formed by bacterial cells that restrict the motion of themselves as the incubation period extends. Though the effect of inoculum age is still unclear, the influence on crystallinity of bacterial cellulose caused by variation of some ordinary culture conditions can be drawn out from data of this work. PMID- 21630053 TI - Monitoring intra-cardiac shunts correction with transpulmonary thermodilution curve: the best is yet to come! PMID- 21630054 TI - Fourth ventricular choroid plexus papilloma. PMID- 21630055 TI - The expression of CXCL12 and CXCR4 in gastric cancer and their correlation to lymph node metastasis. AB - The purpose of this study was to investigate the expression of CXCL12 and its receptor CXCR4 in gastric cancer and to determine their relationship with lymph node metastasis. Fifty patients with pathologically confirmed gastric cancer were analyzed from September 2004 to December 2004. The expression levels of CXCL12 and CXCR4 were examined by immunohistochemical staining in the primary gastric tumor tissues, adjacent normal mucosa tissues, and metastatic lymph nodes and were analyzed along with clinicopathological risk factors, to determine their correlation with the prognosis. Positive staining for CXCL12 and CXCR4 was identified in 90.0 and 80.0% of the primary gastric tumor tissues, respectively, with significantly higher expression intensities observed in the primary gastric tumor tissues than in the adjacent normal mucosa tissues (P < 0.01 and P = 0.01, respectively). Positive staining for CXCL12 and CXCR4 was identified in 94.4 and 91.7% of metastatic lymph nodes, respectively, with significantly higher expression intensities in the metastatic lymph nodes than in the adjacent normal mucosa tissues (P < 0.01 and P = 0.01, respectively). Expression of CXCL12 in the primary gastric tumor tissues was not significantly associated with the clinicopathological characteristics of the tumor or the disease prognosis. However, the intensity of CXCR4 staining in primary tumor tissues was positively related with lymph node metastasis, TNM staging, and disease prognosis (P = 0.04, 0.03, 0.03, respectively). CXCL12 and CXCR4 are related to formation of gastric tumors and lymph node metastasis. Furthermore, the expression of CXCR4 could be used as a biomarker to predict malignant features of gastric cancer. PMID- 21630056 TI - Can patients with metastatic breast cancer be cured after introduction of newer and more effective agents? AB - Unlike early stage breast cancer, metastatic breast cancer (MBC) is generally considered incurable except for a small number of patients with oligometastatic disease. The goal of treatment of MBC should be the prolongation of life and improvement of symptoms and quality of life. The prognoses of patients with MBC, however, have been improved with the introduction of newer, more effective agents. Therefore, the clinical question arises whether MBC patients can be cured with these new therapeutic agents. However, there are a couple of problems in tackling this question, including the duration of follow-up and the presence of strong adjuvant therapy. Firstly, most trials in MBC have a relatively short follow-up; long-term surveillance (>3-5 years) is exceptional, so little is known about the definitive outcome and the exact proportion of long-term survivors. Secondly, most of the patients have received pre- or postoperative adjuvant therapy. The cancer cells at metastatic sites are considered to be relatively resistant to the agents used in metastatic settings. Promisingly, a number of novel therapeutic agents including antibody-drug conjugates, irreversible small molecule HER2-tyrosine inhibitors, and HER2 dimerization inhibitors show promise in the treatment of HER2-overexpressing MBC, as well as PARP-1 [poly(ADP-ribose) polymerase-1] inhibitors for triple-negative breast cancer. PMID- 21630057 TI - TS gene polymorphisms are not good markers of response to 5-FU therapy in stage III colon cancer patients. AB - AIM: Although the predictive and prognostic value of thymidylate synthase (TS) expression and gene polymorphism in colon cancer has been widely studied, the results are inconclusive probably because of methodological differences. With this study, we aimed to elucidate the role of TS gene polymorphisms genotyping in therapy response in stage III colon carcinoma patients treated with 5-FU adjuvant chemotherapy. PATIENTS AND METHODS: 251 patients diagnosed with stage III colon carcinoma treated with surgery followed by 5-FU based adjuvant therapy were selected. The variable number of tandem repeats (VNTR) and the single nucleotide polymorphism (SNP) in the 5'untranslated region of the TS gene were genotyped. RESULTS: There was a positive association between tumor T stage and the VNTR genotypes (p = 0.05). In both univariate and multivariate survival analysis no effects of the studied polymorphisms on survival were found. However, there was an association between both polymorphisms and age. Among patients younger than 60 years, the patients homozygous for 2R seemed to have a better overall survival, whereas among the patients older than 67 this longer survival was seen by the carriers of other genotypes. CONCLUSION: We conclude that the TS VNTR and SNP do not predict response to 5-FU therapy in patients with stage III colon carcinoma. However, age appears to modify the effects of TS polymorphisms on survival. PMID- 21630058 TI - Controversies in RELN/reelin expression in otosclerosis. AB - Several studies have reported a potential genetic association between disease specific single nucleotide polymorphism (SNPs) of RELN and otosclerosis and confirmed RELN expression in human stapes footplates. These are conflicting results, since RELN expression has been attributed exclusively to neural tissues and to odontoblasts. Otosclerosis is a disease of complex bone remodeling disorder, which is limited to the human otic capsule. Genetic predisposition has long been suspected, however, the pathogenesis remained unclear. Ankylotic stapes footplates (n = 85), cortical bone fragments (n = 4), hearing ossicles (n = 2) and human brain tissue specimens (n = 4) were processed to RELN-specific RT-PCR and reelin-specific immunofluorescent assay (IFA). The first group of ankylotic stapes footplates (n = 22) showed a consistent positive reaction against reelin by IFA; however, RELN-specific mRNA could not be detected in the second, RT-PCR group (n = 63). Brain specimens were characterized by robust expression of reelin (n = 2) and RELN-specific mRNA (n = 2). In case of bone-specific controls (n = 6), reelin/RELN expression was excluded obviously. Concerning current observations, RELN gene does not show active expression in adult stapes footplates. Since, the otic capsule surrounds a special neural structure (membranous labyrinth), reelin might play a coordinative role in the early embryonic stage of development. As being a part of the otic capsule, stapes footplate might be characterized by persisting reelin detectability without mRNA expression. Between these conditions, the etiologic role of RELN is questionable in the pathogenesis of otosclerosis. PMID- 21630059 TI - The alternative ear-piercing technique by using superpulsed carbon dioxide laser: a comparative study with spring-loaded gun. AB - Ear piercing (EP) is increasing in popularity among teenagers. Techniques for EP should be selected carefully to prevent possible complications. The purpose of this study is to compare the clinical outcomes of EP techniques between CO(2) laser and spring-loaded gun. This is a prospective and comparative clinical trial. Under local anesthesia, EP was performed on left ear with CO(2) laser (20 watt/single mode) and on right ear with spring-loaded gun. With visual analog scale (VAS, 0-10) and questionnaire, post-operative pain and wound healing status were assessed immediately, 1, 2, 4 and 8 weeks after EP procedure. Fourteen subjects (14/17, 82.4%) completed the whole study. Immediately after the procedure, the level of post-operative pain (VAS) was 3.2 for CO(2) laser and 1.5 for spring-loaded gun (p < 0.05). In CO(2) laser group, pain severity decreased to 0.4, 0.1, 0, and 0, while in spring-loaded gun group, the decrease was only to 0.7, 0.6, 0.3 and 0 at 1, 2, 4, and 8 weeks, respectively. Duration of post operative pain was 3.8 and 17.5 days for CO(2) laser and spring-loaded gun, respectively (p < 0.05). There was no major complication like infection, bleeding or hypertrophic scar. Our study suggests that CO(2) laser is a precise, simple, safe and aseptic technique for EP. It has a lower level and shorter duration of post-operative pain, when compared with spring-loaded gun. Therefore, CO(2) assisted EP is an alternative and feasible technique in our daily clinical practice. The level of evidence: 2b. PMID- 21630060 TI - Controversies about the central compartment in thyroid cancer. Editorial regarding the article "Clinical impact of cervical lymph node involvement and central neck dissection in patients with papillary thyroid carcinoma: a retrospective analysis of 368 cases" by Alexandre Bozec et al. PMID- 21630061 TI - Interactions in the cochlea between air conduction and osseous and non-osseous bone conduction stimulation. AB - Since air-conducted (AC) and clinical (mastoid) bone-conducted (BC) sounds interact in the cochlea (e.g. pitch, cancellation, masking, beats), it has been thought that both AC and BC stimulations lead to a mechanical wave in the cochlea. However, there are also "non-osseous" forms of BC, i.e. auditory sensation produced when the clinical bone vibrator is applied to "non-osseous" soft tissue sites. In the present study, such "non-osseous" sites were identified (e.g. eye, cheek, neck) and they interacted with AC and osseous BC (pitch matching, beats, masking), indicating that all of these forms of auditory stimulation converge in the cochlea, producing the same pattern of mechanical activity, leading to their interactions. PMID- 21630062 TI - Management of ear lobule keloids using 980-nm diode laser. AB - The objective of the study was to evaluate the 980-nm diode laser in conjunction with corticosteroids in the treatment of ear lobule keloids. Several methods have been described for the treatment of keloid scars, but none of them have been 100% successful. Advances in laser techniques have enabled surgeons to define the most appropriate lasers for use in the treatment of different scar types. The diode laser pulses are delivered interstitially in a single repeated mode in non overlapping sites using a bare optical fiber, followed by intralesional triamcinolone acetonide injection. The number of sessions varies between two to five for the management of more than 75% of keloid size, with a total success rate of 75% and no recurrence in the follow-up of 12 months. The technique used proved to be effective in the treatment of ear lobule keloids. PMID- 21630063 TI - Novel composite implant in craniofacial bone reconstruction. AB - Bioactive glass (BAG) and polymethyl methacrylate (PMMA) have been used in clinical applications. Antimicrobial BAG has the ability to attach chemically to surrounding bone, but it is not possible to bend, drill or shape BAG during the operation. PMMA has advantages in terms of shaping during the operation, but it does not attach chemically to the bone and is an exothermic material. To increase the usefulness of BAG and PMMA in skull bone defect reconstructions, a new composite implant containing BAG and PMMA in craniofacial reconstructions is presented. Three patients had pre-existing large defects in the calvarial and one in the midface area. An additive manufacturing (AM) model was used preoperatively for treatment planning and custom-made implant production. The trunk of the PMMA implant was coated with BAG granules. Clinical and radiological follow-up was performed postoperatively at 1 week, and 3, 6 and 12 months, and thereafter annually up to 5 years. Computer tomography (CT) and positron emission tomography (PET-CT) were performed at 12 and 24 months postoperatively. Uneventful clinical recovery with good esthetic and functional outcome was seen. CT and PET-CT findings supported good clinical outcome. The BAG-PMMA implant seems to be a promising craniofacial reconstruction alternative. However, more clinical experience is needed. PMID- 21630064 TI - [Pain and anesthesiology : aspects of the development of modern pain therapy in the twentieth century]. AB - The connection between the development of anesthesiology and pain therapy in the twentieth century is close. The optimistic idea to overcome pain by using general anesthesia derives from the nineteenth century. Treatment of nonsurgical pain remained in the background for a long time and innovations in pain medicine did not improve the insufficient care for patients with postoperative pain. Therapy of chronic pain was mainly surgical and the extreme of this surgical approach was psychosurgery. In the years following World War II leucotomy and lobotomy were established as methods to separate the psychological processing of pain from the experience of pain. Meanwhile, the French "pain surgeon" Rene Leriche elaborated a theory of pain where chronic pain was no longer seen as a symptom but as a "douleur-maladie", a pain disease. His theory was considered on various occasions but did not gain acceptance before the 1950s. Research in anesthesiology, such as that conducted by the American scientist Henry Beecher separated psyche and physiology with respect to pathological pain. This was contrasted by the approach of clinical anesthesia to pain therapy, which was based on regional anesthesia. The first "pain clinics" were "nerve block clinics". John Bonica, a regional anesthesiologist, extended the framework of pain therapy by introducing multidisciplinary teamwork into the therapy of chronic pain. From today's viewpoint his 1953 monograph The Management of Chronic Pain is a milestone in the development of modern pain therapy. However, Bonica's work did not attain major importance until 1960 when he was appointed to a newly established chair. Gradually, chronic pain was recognized as an independent illness and differentiated as such from acute pain. In 1965 the gate control theory by Melzack and Wall offered a possible explanation for the mechanisms of chronic pain. By the end of the 1970s the spectrum was extended to the biopsychosocial approach which was foremost developed by the American psychiatrist George Engel, defined chronic pain as an illness rather than a disease. Concurrently, the radical behaviorism of the late 1960s affected both the therapy of chronic and of acute pain. Based on this theory, patient-controlled analgesia (PCA) was introduced in the 1970s and 1980s. Acute pain services (APS) in hospitals, were developed beginning in the 1980s using the continuous release of opioids. Regional anesthesia played a greater role than general anesthesia in developing pain therapy in the twentieth century and paved the way for pain therapy. The restriction to nerve blocks in pain centers was overcome by the expansion of theoretical foundations beyond the framework of anesthesiology. Impulses from psychology and psychosomatic medicine were crucial. The evolution of cancer pain therapy was distinct from non-cancer pain therapy. PMID- 21630065 TI - Variability in the clinical management of fatty acid oxidation disorders: results of a survey of Canadian metabolic physicians. AB - INTRODUCTION: There is little robust empirical evidence on which to base treatment recommendations for fatty acid oxidation disorders. While consensus guidelines are important, understanding areas where there is a lack of consensus is also critical to inform priorities for future evaluative research. METHODS: We surveyed Canadian metabolic physicians on the treatment of medium-chain acyl-CoA dehydrogenase (MCAD) deficiency, very long-chain acyl-CoA dehydrogenase (VLCAD) deficiency, long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency, and mitochondrial trifunctional protein (MTP) deficiency. We ascertained physicians' opinions on the use of different interventions for the long-term management of patients as well as for the management of acute illness, focusing on identifying interventions characterized by high variability in opinions. We also investigated factors influencing treatment decisions. RESULTS: We received 18 responses (response rate 45%). Participants focused on avoidance of fasting and increased meal frequency as interventions for the management of MCAD deficiency. For the long-chain disorders, avoidance of fasting remained the most consistently endorsed intervention, with additional highly endorsed treatments differing for VLCAD versus LCHAD/MTP deficiency. L-carnitine supplementation and restriction of dietary fat were characterized by high variability in physicians' opinions, as were several interventions specific to long-chain disorders. Social factors and patient characteristics were important influences on treatment decisions. CONCLUSIONS: Based on our findings we suggest that high priority treatments for rigorous effectiveness studies could include L-carnitine supplementation (MCAD and LCHAD/MTP deficiencies), restriction of dietary fat, and, for the long-chain disorders, feeding practices for breastfed infants and the use of various supplements (essential fatty acids, carbohydrates, cornstarch, multivitamins). PMID- 21630066 TI - Hemolytic disease of the newborn- anti c antibody induced hemolysis. AB - Hemolytic disease in the newborn, as a cause of early jaundice, is not uncommon. This is mostly due to Rh (D), ABO incompatibility and rarely due to other minor blood group incompatibility. The authors report two cases of Rh anti c isoimmunization presenting as significant early neonatal jaundice within the 20 h of life. Both the babies were treated with intensive phototherapy. One baby underwent exchange transfusion and the other required packed cell transfusion for anemia. PMID- 21630067 TI - Acute myeloid leukemia (AML-M2) with translocation (8;21) (q22;q22) and abnormal eosinophilic precursors in the bone marrow--a case report. AB - The translocation (8;21)(q22;q22) is frequently associated with M2 subtype of AML. The authors herein present a case of AML-M2 in a nine-year-old boy without hepatosplenomegaly, lymphadenopathy or any bleeding diathesis. Bone marrow examination revealed high number of eosinophilic precursors (60%) among the total nucleated bone marrow cells. Cytogenetic study with G- banding method showed 46, XY, t (8;21)(q22;q22). The morphological abnormalities in eosinophils observed in AML suggested that eosinophils may be a part of leukemic process. PMID- 21630068 TI - Wiedemann-Rautenstrauch syndrome: first Indian case. AB - Wiedemann-Rautenstrauch syndrome, also known as neonatal progeroid syndrome (NPS), is an extremely rare genetic disorder characterised by an aged appearance at birth. About thirty cases have been reported in the literature. The authors report first Indian baby with Wiedemann-Rautenstrauch syndrome with the lowest birth weight documented in such a patient, who is still surviving at 24 months. This ethnicity has not been reported previously in literature. PMID- 21630069 TI - Characteristics of pediatric scrub typhus during an outbreak in the North Eastern region of India: peculiarities in clinical presentation, laboratory findings and complications. AB - OBJECTIVE: To identify and report a recent outbreak of scrub typhus cases recorded from October 2009 to January 2010 in the state of Meghalaya, India. METHODS: The case sheets of all the children were retrieved and reviewed retrospectively. Twenty four (24) patients, who were both clinically and serologically confirmed as scrub typhus cases were included in the study. RESULTS: Fever for more than 1 wk duration was the most common manifestation (100%) followed by splenomegaly (45.8%), eschar (41.7%), cough (37.5%), hepatomegaly (33.3%), headache and body ache (25%), pain abdomen (25%), vomiting (20.8%), altered sensorium (16.6%), seizures (12.5%) lymphadenopathy (12.5%), and loose stools (8.3%). Meningoencephalitis was the most common complication (29.2%) followed by pneumonia (16.3%) and subconjunctival hemorrhage (8.3%).Cortical blindness, septic shock, peritonitis, myocarditis with CCF, pancytopenia, acute renal failure, coagulopathy, prolonged oxygen dependency and urinary tract infection (UTI) were found in one of each case. Hyponatremia (66.7%), elevated liver enzymes without significant rise of bilirubin (58.3%), hypoalbuminemia (52.2%) and thrombocytopenia (26%) were the other significant laboratory findings. Patients were treated with chloramphenicol, doxycycline and azithromycin. There was no mortality. CONCLUSIONS: This is the first outbreak report from the north eastern region of India with varied clinical presentations, laboratory investigations and complications. Weil Felix test still remains fruitful for diagnosing this disease in a resource limited set up. PMID- 21630070 TI - Movement assessment of infants as a predictor of one year neuromotor outcome in very low birth weight infants. AB - OBJECTIVE: To predict neuromotor outcome in Very Low Birth Weight infants at 1 year using Movement Assessment of Infants scale at 4 and 8 months of corrected age. METHODS: This hospital based prospective observational study consecutively recruited 45 infants with birth weight <= 1,500 g and admitted to neonatal intensive care unit. The infants were evaluated at 4 and 8 months of corrected age using Movement Assessment of Infants scale and then a follow up evaluation was done at 1 year of corrected age with a neuropschycological tool, Bayley Scale of Infant Development-II and psychomotor development index was calculated to identify the neuromotor outcome. RESULTS: For predicting neuromotor outcome on the Bayleys motor scale at 1 year, the sensitivity of Movement Assessment of Infants for 4-month was 70%, specificity 83.3%, positive predictive value 58.3% and negative predictive value was 89.3% for risk points >= 10. The sensitivity of Movement Assessment of Infants for 8-month was 80%, specificity 93.3%, positive predictive value 80% and negative predictive value was 93.3% for risk points >= 10 .The correlations of the Movement Assessment of Infants total risk scores and categorical risk scores for muscle tone, primitive reflex, automatic reactions and volitional movement at 4 and 8 months with Bayley motor scale at 1 year were highly significant (p < 0.0000). CONCLUSIONS: Movement Assessment of Infants is an effective clinical tool to identify neuromotor abnormalities, as the best predictive values for neuromotor outcome at 1 year in Very Low Birth Weight infants were obtained at 8-months and the 4-month Movement Assessment of Infants had high specificity and acceptable sensitivity. PMID- 21630071 TI - Megaloblastic anemia--a rare cause. AB - A 2- year- old boy presented with non responsive megaloblastic anemia, growth failure and developmental delay. Blood levels of B(12), folic acid and iron were normal. Tandem mass spectroscopy for common inborn errors of metabolism did not reveal any abnormality. There was an increased excretion of orotic acid in urine. The authors report this as a rare cause of megaloblastic anemia. PMID- 21630072 TI - The role of hemoglobin variant replacement in retinopathy of prematurity. AB - OBJECTIVE: To conduct tests of relationships between different factors that could influence the course of retinopathy of prematurity (ROP) and ROP, particularly the role of hemoglobin variant replacement in adult blood transfusions. METHODS: A retrospective, observational study of 83 infants born between 23 and 34 wks gestation was conducted. RESULTS: The infants without ROP, with 1 and 2 stage of ROP and with >= 3 stage of ROP received Q 28 (12-134); 51 (14-149); 156 (38-244) ml/kg of transfused blood, respectively, and the factor Qt was 1,545 (560 10,045); 3,093 (614-13,419); 11,907 (1,288-20,638) (ml/kg).day, respectively. For the same groups MCV(35) (mean cell volume at the arbitrary time of the 35(th) wk post-conception) was 92.3 (82.9-110.5); 91.0 (79.3-101.4); 87.1 (80.2-94.8) fl, respectively, and factor P(MCV)/t was 99.5 (89.2-108.8); 96.3 (84.6-106.3); 90.7 (85.3-96.5) fl, respectively. There is high influence on the stage of ROP of the amount of transfused blood and MCV, both with or without the time factor. The statistical differences between P(MCV)/t were more significant than the differences between MCV(35), for different stages of ROP. CONCLUSIONS: The influence of the time factor on the statistical differences of MCV but not on the amount of transfused adult blood suggests that HbF - HbA replacement may play a role in ROP development. PMID- 21630073 TI - Refractory anemia as a presenting feature of microscopic polyangiitis: a rare vasculitis in children. AB - Microcytic anemia refractory to usual supplementation is an important clue to an alternative diagnosis. Accompanying pulmonary and renal involvement during the disease course suggests a multisystem disease. Small-vessel vasculitis should be suspected in any patient who presents with a multisystem disease that is not caused by an infectious or malignant process. Among these, antineutrophilic cytoplasmic antibodies (ANCA)-associated vasculitis is a distinct subclass involving anti-neutrophil cytoplasmic antibody (ANCA) as the common pathogenesis. Microscopic polyangiitis is a rare form of such vasculitis in children characterized by pulmonary-renal syndrome with pauci-imune rapidly progressive glomerulonephritis and a poor outcome. PMID- 21630074 TI - Psychopathology, traumatic life events, and coping skills among patients attending a primary-care adolescent clinic. AB - OBJECTIVE: To compare the type of life events experienced and coping styles used by adolescents with and without psychopathology, attending a primary-care adolescent clinic. METHODS: One hundred adolescents with and without psychopathology attending a drop-in adolescent clinic in a tertiary-care teaching hospital were recruited. Face-to-face interview used Child Behaviour Checklist, Life Event Scale, Coddington's life event scale, Impact of Event Scale and Modified Jalowiec coping scale as measures after getting written, informed consent from the primary care-giver and verbal assent from the adolescents. Bivariate and multivariate comparisons were done between the groups appropriately. RESULTS: Adolescents with psychopathology had experienced more parental fights, increased arguments with parents, increased arguments between parents, serious illness requiring hospitalization of the adolescent. The intrusive symptoms of PTSD were noted more than avoidant symptoms among those adolescents with life events. Confrontative, emotive and optimistic coping styles were most often used in adolescent with psychopathology. CONCLUSIONS: In India, adolescents with psychopathology attending a primary care clinic have significant life events and different coping styles. Therefore, adolescents with psychopathology in this setting should be screened for life events as well as dysfunctional coping styles and given appropriate intervention. PMID- 21630075 TI - Protein and micronutrient supplementation in complementing pubertal growth. AB - For several reasons, adolescence is considered as a nutritionally critical period of life. The dramatic increase in physical growth and development puts greater pressure on the need for nutrients, as it is during this period when adolescents experience a weight gain equivalent to 65% of their weight at the beginning of the period or 40% of their final weight, and a height gain equivalent to 15% of their adult height. The requirement of some of the nutrients is as high as, or higher in adolescents than in any other age groups. Moreover, adolescence is a time where the life-style and food habits are influenced by socio-cultural factors resulting in an imbalance in nutrient intake and nutrient inadequacy can result in delayed sexual maturation and can arrest or slow linear growth. With the increasing prevalence of hidden malnutrition among Indian children, adolescence can be the second opportunity to catch up growth and cover the deficits suffered during childhood and to meet the demands of physical and cognitive growth and development, provide adequate stores of energy for illnesses and pregnancy, and prevent adult onset of nutrition-related diseases. Adolescents should be recognized as a priority target group as an integral part of health promotion and optimal nutrition through balanced diet and nutrient supplementation should be promoted in conjunction with healthy eating habits and physical activity. PMID- 21630076 TI - Approach to a child with breathing difficulty. AB - Breathing difficulty and respiratory distress is the most common cause of admission to the Pediatric Emergency. Respiratory distress presents as altered breathing pattern, forced breathing efforts or obstructed breathing, and chest indrawing; respiratory failure is defined as paCO(2) >50 mmHg (inadequate ventilation) and/or a paO(2) < 60 mmHg (inadequate oxygenation). Rapid assessment is aimed to ascertain adequacy of airway patency, breathing, and circulation. Immediate care is directed at (a) restoration of airway patency- by positioning (head tilt -chin lift), cleaning the oropharynx, and/or insertion of oropharyngeal airway; (b) supporting breathing- with high flow oxygen and assisted ventilation (with bag and mask or endotracheal intubation and ventilation), and (c) restoration of circulation- using fluid boluses and inotropes, if necessary. Immediate specific management may require endotracheal intubation/tracheostomy for upper airway obstruction; needle thoracotomy and drainage of pneumothorax; and first dose of antibiotic for febrile children. Thereafter meticulous history, focused physical examination, and specific laboratory/radiological investigations are undertaken to identify the underlying cause. At the end of this, one should be able to categorize the child to one of the following: (a) upper airway obstruction, (b) pneumonia (syndrome of cough, fever and breathing difficulty), (c) lower airway obstruction, (d) slow or irregular breathing without pulmonary signs, and (e) respiratory distress with cardiac findings, to initiate specific treatment. Further respiratory support by Continuous Positive Airways Pressure (CPAP) and mechanical ventilation may be required in some cases. All children with respiratory distress must be monitored for early detection of worsening/complications, assessment of response to therapy and rapid documentation of clinical state. PMID- 21630077 TI - Integrating the life course perspective into a local maternal and child health program. AB - For many decades, early access to prenatal care has been considered the gold standard for improving birth outcomes. In Contra Costa County, a diverse urban and suburban county of over one million people in the San Francisco Bay Area, the Family Maternal and Child Health Programs of Contra Costa Health Services (CCHS) have seen high rates of early entry into prenatal care since 2000. Yet despite our best efforts to increase access to quality prenatal care, our rates of low birth weight and infant mortality, especially among African Americans, continue to be high. When we were introduced to the Life Course Perspective in 2003 as an organizational framework for our programmatic activities, we recognized that emerging scientific evidence in the literature demonstrated the importance of social and environmental factors in determining health and health equity, and supported a general impression in the field that prenatal care was not enough to improve birth outcomes. The Life Course Perspective suggests that many of the risk and protective factors that influence health and wellbeing across the lifespan also play an important role in birth outcomes and in health and quality of life beyond the initial years. In this article, we describe the Life Course Perspective and how one local Maternal and Child Health Program adopted and adapted this paradigm by creating and launching a Life Course Initiative to guide our programs and services. The Life Course Initiative implemented by CCHS is designed to reduce inequities in birth outcomes, improve reproductive potential, and change the health of future generations by introducing a longitudinal, integrated, and ecological approach to implementing maternal and child health programs. PMID- 21630078 TI - Bioproduction of hydrolytic enzymes using apple pomace waste by A. niger: applications in biocontrol formulations and hydrolysis of chitin/chitosan. AB - Studies were carried out for beta-glucosidase production using apple pomace (AP) in solid-state fermentation using 2(4) factorial design and response surface methodology. The influence of four independent variables including initial moisture level and inducers [veratryl alcohol (VA), lactose (LAC) and copper sulfate (CS)] was studied. The experimental design showed that initial moisture level had significant negative effect on the response. Higher beta-glucosidase activity of 64.18 IU/gram fermented substrate (gfs) was achieved in solid-state tray fermentation with optimum conditions having initial moisture level 55% (v/w), pH 4.5, 2 mM/kg VA, 2% (w/w) LAC and 1.5 mM/kg CS concentration, respectively,. The non-specific chitinase 70.28 +/- 6.34 IU/gfs and chitosanase activities 60.18 +/- 6.82 to 64.20 +/- 7.12 IU/gfs were observed. The study demonstrated that AP can be potentially used for the beta-glucosidase production by Aspergillus niger. Moreover, beta-glucosidase can be used for the hydrolysis of chitin/chitosan to depolymerized products and in the formulation of biocontrol agents for enhanced entomotoxicity levels. PMID- 21630079 TI - Plasma citrulline as marker of bowel adaptation in children with short bowel syndrome. AB - PURPOSE: The aim of this study is to determine if prospective determinations of citrulline could be predictive of the bowel adaptation in children with short bowel syndrome (SBS). METHODS: Between March 2005 and March 2010, we prospectively included 28 SBS patients on parenteral nutrition. The citrulline and the enteral intake determinations were scheduled at the inclusion and at 6 month intervals. We assessed the correlation between citrulline and bowel length as well as enteral caloric intake, longitudinal trend of citrulline and association between patients characteristics according to the course of bowel adaptation. RESULTS: Citrulline significantly correlated with the residual duodenum-jejunum length (r (2) = 0.22, P = 0.0113) and with enteral intake (r (2) = 0.20, P = 0.016 and r (2) = 0.48, P = 0.0001). Baseline citrulline at the cutoff >10 MUmol/L and a longitudinal increase >25% provided a weak association with bowel adaptation (likelihood ratio (LR), 2.6 and 2.4, respectively), unlike residual small bowel length >=20 cm and the presence of >50% of the colon (LR, 10 and 6, respectively). CONCLUSIONS: Citrulline seems to be a powerful biomarker of the intestinal function, showed by the correlation with the residual duodenum jejunum length and the enteral absorption, but not of its prospective changes during the bowel adaptation process. Future studies may be necessary to confirm this finding. PMID- 21630080 TI - State of the art: surgery for endemic goiter--a plea for individualizing the extent of resection instead of heading for routine total thyroidectomy. AB - BACKGROUND: During the past 150 years of nodular goiter surgery, the pendulum has been swinging from limited to more extensive forms of thyroidectomy and all the way back reflecting the challenges of striking a balance between surgical morbidity and disease recurrence. PURPOSE: This article aimed at providing guidance for individualizing decisions using a risk-oriented surgical approach to endemic goiter based on literature review and personal experience. DISCUSSION: The following arguments favor total thyroidectomy: (a) Endemic goiter involves the entire thyroid gland; (b) Increasing standardization and specialization supported by better visualization, surgical devices, and intraoperative parathyroid hormone assays have decreased surgical morbidity; (c) One third of goiter patients require completion thyroidectomy for incidental thyroid cancer; (d) Recurrent goiter is frequently seen on ultrasonography after subtotal thyroidectomy; (e) Thyroid hormone replacement is well tolerated and inexpensive. Important counter-arguments include: (a) Restoration of iodine sufficiency does not reverse nodular goiter nor can the growth of individual nodules be predicted; (b) To gather the annual case load necessary to achieve improved outcomes, surgeons need to "super-specialize", which may not be viable globally; (c) Many incidental cancers are detectable through high-resolution ultrasonography, fine needle aspiration cytology, and frozen section during thyroidectomy; (d) Not all recurrent goiters require reoperations; (e) Thyroid hormone replacement is not available and affordable everywhere. CONCLUSION: The higher surgical morbidity associated with total thyroidectomy, notably recurrent laryngeal nerve palsy and hypoparathyroidism, calls for individualizing the extent of resection for endemic goiter as a new standard of care instead of heading for routine total thyroidectomy. PMID- 21630081 TI - Depressive symptoms in the Belgian population: disentangling age and cohort effects. AB - OBJECTIVE: Although the association between age and depression has been previously demonstrated, uncertainty remains because of the confounding relationship existing between age and cohort. A study by Yang (J Health Soc Behav 48(1):16, 2007) has evidenced important cohort effects and age-by-cohort interactions in depressive symptoms among US citizens. A crucial limitation, however, is that this study confines itself to elderly population. The objective of the present study is to bring further clarification to the association between age, cohort membership and depressive symptoms, by analyzing a sample with a wider age range. METHODS: The Panel Study of Belgian Households is a prospective longitudinal survey, following adults ages 25-74, annually from 1992 to 2002. Missing data were replaced using multiple imputation, allowing for a complete dataset (N = 7,000) at each wave. Respondents were classified into one of five birth cohorts: 1918-1927; 1928-1937; 1938-1947; 1948-1957; 1958-1967. Frequency of depressive symptoms was reported using a modified version of the Health and Daily Living form. Growth curve modeling was used to determine the effect of age and cohort on depression trajectory. RESULTS: All cohorts differed significantly from one another, with recent cohorts always obtaining the highest mean HDL depression score. The intensity of depressive symptoms increases linearly with age, but significant age-by-cohorts interactions were detected, indicating that the relationship between age and depression varies across cohorts. No evidence of a WW2 effect was found. CONCLUSION: The association between age and depression has to take cohort membership into account. Cohort replacement effects explain the increase in depression in Belgium. PMID- 21630082 TI - Measuring attitude toward social health insurance. AB - In order to understand the health care system a country chooses to adopt or the health care reform a country decides to undertake, one must first be able to measure a country's attitude toward social health insurance. Our primary goal was to develop a construct that allows us to measure this "attitude toward social health insurance". Using a sample of 724 students from the People's Republic of China, Germany, and the United States and an initial set of sixteen items, we extract a scale that measures the basic attitude toward social health insurance in the three countries. The scale is internally consistent in each of the three countries. A secondary factor labeled "government responsibility" is marginally consistent for the total sample and for the German sample. German respondents have the most favorable attitude toward social health insurance, followed by China, and then the United States. Chinese respondents have the most favorable attitude toward government responsibility in health insurance. The scale developed here can be used to further investigate and understand which health care system will succeed and which will fail in a given country, which is important from both a political and an economic perspective. PMID- 21630083 TI - An engineered cysteine-modified diabody for imaging activated leukocyte cell adhesion molecule (ALCAM)-positive tumors. AB - PURPOSE: The purpose of this study was to generate and evaluate a positron emission tomography (PET) radiotracer targeting activated leukocyte cell adhesion molecule (ALCAM). PROCEDURES: A human anti-ALCAM single chain variable fragment was reformatted to produce a covalent dimer, termed a cys-diabody (CysDb). Purified CysDb was characterized by gel electrophoresis and size exclusion chromatography, and immunoreactivity was assessed by flow cytometry and immunofluorescence. Targeting and imaging of ALCAM-positive tumors using (64)Cu DOTA-CysDb were evaluated in mice bearing human pancreatic adenocarcinoma xenografts (HPAF-II or BxPC-3). RESULTS: CysDb binds specifically to ALCAM positive cells in vitro with an apparent affinity in the range of 1-3 nM. MicroPET images at 4 h showed specific targeting of positive tumors in vivo, a finding confirmed by biodistribution analysis, with positive/negative tumor ratios of 1.9 +/- 0.6 and 2.4 +/- 0.6, and positive tumor/blood ratios of 2.5 +/- 0.9 and 2.9 +/- 0.6 (HPAF-II and BxPC-3, respectively). CONCLUSIONS: Successful imaging with (64)Cu-DOTA-CysDb in animal models suggests further investigation of ALCAM as an imaging biomarker is warranted. PMID- 21630085 TI - Regulation of chemosensitivity and migration by clusterin in non-small cell lung cancer cells. AB - PURPOSE: In terms of drug resistance, cancer cells usually benefit from high clusterin (CLU) expression on chemotherapy. In contrast, CLU expression has been found to be a favorable prognostic factor in lung cancer patients. The aims of this study are to determine the association between CLU expression and chemotherapeutic sensitivity and the potential role of CLU in migration in human non-small-cell lung cancer (NSCLC) cell lines. METHODS: The levels of clusterin in NSCLC cell lines were altered by short hairpin RNA interference (shRNAi) and overexpression on chemosensitivity assay. Migratory ability of these cell lines was also investigated. RESULTS: H1355 cells with the highest level of CLU demonstrated the lowest sensitivities to Adriamycin (ADR), docetaxel (DOC), and gemcitabine (GEM) treatment. Inhibition of CLU expression in H1355 cells resulted in higher chemosensitivities. When CLU was stably overexpressed in A549 and H1299 cells, only the chemosensitivity to ADR was reduced. The migratory ability of CLU overexpressing cells significantly decreased. Moreover, MMP2 transcription was inhibited in CLU-overexpressing H1299 cells. These results indicated lower metastatic potential for cancer cells with high CLU level. CONCLUSION: Lung cancer cells with high level of CLU have reduced chemosensitivity. High level of CLU may result in migratory inhibition and thus favorable prognosis in lung cancer. PMID- 21630086 TI - Further characterisation of the cellular activity of the DNA-PK inhibitor, NU7441, reveals potential cross-talk with homologous recombination. AB - PURPOSE: Inhibition of DNA repair is emerging as a new therapeutic strategy for cancer treatment. One promising target is DNA-PK, a pivotal kinase in double strand break repair. The purpose of this study was to further characterise the activity of the DNA-PK inhibitor NU7441, giving some new insights into the biology of DNA-PK. METHODS: We used NU7441, a potent DNA-PK inhibitor, to evaluate potential pharmacodynamic markers of DNA-PK inhibition, inhibition of DNA repair and chemo- and radio-potentiation in isogenic human cancer cells proficient (M059-Fus1) and deficient (M059 J) in DNA-PK. RESULTS: NU7441 strongly inhibited DNA-PK in cell lines (IC(50) = 0.3 MUM) but only weakly inhibited PI3 K (IC(50) = 7 MUM). The only available anti-phospho-DNA-PK antibody also recognised some phosphoprotein targets of ATM. NU7441 caused doxorubicin- and IR-induced DNA DSBs (measured by gamma-H2AX foci) to persist and also slightly decreased homologous recombination activity, as assessed by Rad51 foci. Chemo- and radio potentiation were induced by NU7441 in M059-Fus-1, but not in DNA-PK-deficient M059 J cells. DNA-PK was highly expressed in a chronic lymphocytic leukaemia sample but undetectable in resting normal human lymphocytes, although it could be induced by PHA-P treatment. In K652 cells, DNA-PK expression was not related to cell cycle phase. CONCLUSION: These data confirm NU7441 not only as a potent chemo- and radio-sensitiser clinical candidate but also as a powerful tool to study the biology of DNA-PK. PMID- 21630084 TI - Sensory integration, sensory processing, and sensory modulation disorders: putative functional neuroanatomic underpinnings. AB - This paper examines conditions that have variously been called sensory integration disorder, sensory processing disorder, and sensory modulation disorder (SID/SPD/SMD). As these conditions lack readily and consistently agreed upon operational definitions, there has been confusion as to how these disorders are conceptualized. Rather than addressing various diagnostic controversies, we will instead focus upon explaining the symptoms that are believed to characterize these disorders. First, to clarify the overall context within which to view symptoms, we summarize a paradigm of adaptation characterized by continuous sensorimotor interaction with the environment. Next, we review a dual-tiered, integrated model of brain function in order to establish neuroanatomic underpinnings with which to conceptualize the symptom presentations. Generally accepted functions of the neocortex, basal ganglia, and cerebellum are described to illustrate how interactions between these brain regions generate both adaptive and pathological symptoms and behaviors. We then examine the symptoms of SID/SPD/SMD within this interactive model and in relation to their impact upon the development of inhibitory control, working memory, academic skill development, and behavioral automation. We present likely etiologies for these symptoms, not only as they drive neurodevelopmental pathologies but also as they can be understood as variations in the development of neural networks. PMID- 21630087 TI - Three-dimensional vibration-induced vestibulo-ocular reflex identifies vertical semicircular canal dehiscence. AB - Vertical semicircular canal dehiscence (VSCD) due to superior canal dehiscence (SCD) or posterior canal dehiscence (PCD) of the temporal bone causes vestibular and cochlear hypersensitivity to sound. This study aimed to characterize the vibration-induced vestibulo-ocular reflex (ViVOR) in VSCD. ViVORs in one PCD and 17 SCD patients, confirmed by CT imaging reformatted in semicircular canal planes, were measured with dual-search coils as binocular three-dimensional eye rotations induced by skull vibrations from a bone oscillator (B71-10 ohms) at 7 ms, 500 Hz, 135-dB peak-force level (re: 1 MUN). The ViVOR eye rotation axes were computed by vector analysis and referenced to known semicircular canal planes. Onset latency of the ViVOR was 11 ms. ViVOR from VSCD was up to nine times greater than normal. The ViVOR's torsional rotation was always contraversive torsional (the eye's upper pole rotated away from the stimulated ear), i.e. its direction was clockwise from a left and counterclockwise from a right VSCD, thereby lateralizing the side of the VSCD. The ViVORs vertical component distinguishes PCD from SCD, being downwards in PCD and upwards in SCD. In unilateral VSCD, the ViVOR eye rotation axis aligned closest to the dehiscent vertical semicircular canal axis from either ipsilateral or contralateral mastoid vibrations. However, in bilateral VSCDs, the ViVOR eye rotation axis lateralized to the ipsilateral dehiscent vertical semicircular canal axis. ViVOR was evoked in ossicular chain dysfunction, even when air-conducted click vestibulo-ocular reflex (VOR) was absent or markedly reduced. Hence, ViVOR could be a useful measurement to identify unilateral or bilateral VSCD even in the presence of ossicular chain dysfunction. PMID- 21630088 TI - Genetic effects of adiponectin single nucleotide polymorphisms on the clustering of metabolic risk factors in young Korean adults. AB - Little is known if lifestyle-related risk factors modulate the adiponectin genetic effects on its outcome phenotypes. The aims of the study were to investigate whether the association between the adiponectin gene two SNPs (+45T>G, rs2241766 and +276G>T, rs1501299) and a clustering of metabolic risk factors is modified by both cardiorespiratory fitness (CRF) and insulin resistance in a study sample of 1,622 young Korean adults (941 men and 681 women, mean age 22.9 +/- 2.4 years). The clustering of metabolic risk factors was defined as a sum of Z scores for waist circumference (WC), blood pressure (BP), triacylglycerols (TAG), high-density lipoprotein cholesterol, and fasting glucose. With respect to SNP +45T>G, the TT genotype had significantly higher values for BMI, WC, systolic BP, TAG, insulin, and a clustered risk score than did the TG+GG genotype. The genetic effect of SNP +45 on the clustered risk score remained significant even after controlling for age, sex, and smoking (P = 0.019). However, the genetic impact was no longer significant when additionally controlling for CRF (P = 0.097) and fasting insulin (P = 0.181), respectively. With respect to SNP +276G>T, the GT+TT genotypes had significantly higher values for BMI and TAG than did the TT genotype. In summary, the present findings suggest that the SNPs at position +45 and +276 are associated with several of metabolic risk factors; however, the genetic effect of SNP +45T/G variant on the clustered risk score is modulated by both CRF and insulin resistance. PMID- 21630089 TI - A non-negative matrix factorization framework for identifying modular patterns in metagenomic profile data. AB - Metagenomic studies sequence DNA directly from environmental samples to explore the structure and function of complex microbial and viral communities. Individual, short pieces of sequenced DNA ("reads") are classified into (putative) taxonomic or metabolic groups which are analyzed for patterns across samples. Analysis of such read matrices is at the core of using metagenomic data to make inferences about ecosystem structure and function. Non-negative matrix factorization (NMF) is a numerical technique for approximating high-dimensional data points as positive linear combinations of positive components. It is thus well suited to interpretation of observed samples as combinations of different components. We develop, test and apply an NMF-based framework to analyze metagenomic read matrices. In particular, we introduce a method for choosing NMF degree in the presence of overlap, and apply spectral-reordering techniques to NMF-based similarity matrices to aid visualization. We show that our method can robustly identify the appropriate degree and disentangle overlapping contributions using synthetic data sets. We then examine and discuss the NMF decomposition of a metabolic profile matrix extracted from 39 publicly available metagenomic samples, and identify canonical sample types, including one associated with coral ecosystems, one associated with highly saline ecosystems and others. We also identify specific associations between pathways and canonical environments, and explore how alternative choices of decompositions facilitate analysis of read matrices at a finer scale. PMID- 21630090 TI - Analysis of rpoS and bolA gene expression under various stress-induced environments in planktonic and biofilm phase using 2(-DeltaDeltaCT) method. AB - Genetic adaptation is one of the key features of Escherichia coli (E. coli) that ensure its survival in different hostile environments. E. coli seems to initiate biofilm development in response to specific environmental cues. A number of properties inherent within bacterial biofilms indicate that their gene expression is different from that of planktonic bacteria. Two of the possible important genes are rpoS and bolA. The rpoS gene has been known as the alternative sigma (sigma) factor, which controls the expression of a large number of genes, which are involved in responses to a varied number of stresses, as well as transition to stationary phase from exponential form of growth. Morphogene bolA response to stress environment leads to round morphology of E. coli cells, but little is known about its involvement in biofilms and its development or maintenance. The purpose of this study was to understand and analyse the responses of rpoS and bolA gene to sudden change in the environment. In this study, E. coli K-12 MG1655, rpoS, and bolA mutant strains were used and gene expression was studied. Results show that both genes contribute to the ability to respond and adapt in response to various types of stresses. RpoS response to various stress environments was somehow constant in both the planktonic and biofilm phases, whereas bolA responded well under various stress conditions, in both planktonic and biofilm mode, up to 5-6-fold change in the expression was noticed in the case of pH variation and hydrogen peroxide stress (H(2)O(2)) as compared with rpoS. PMID- 21630091 TI - Urinary-type plasminogen activator receptor (uPAR) modulates oral cancer cell behavior with alteration in p130cas. AB - Oral cavity cancer is among the most frequently diagnosed cancers worldwide and urinary-type plasminogen activator receptor (uPAR) is clinically associated with more invasive tumors and enhanced lymph node metastasis. We seek to further elucidate the mechanism of by which uPAR promotes cell aggressiveness in the unique context of oral squamous cell carcinoma (OSCC). The contribution of uPAR expression to aggressive cellular behavior of OSCC was examined using in vitro cellular models wherein the expression of uPAR was manipulated and in a human OSCC tissue microarray. Results show altered adhesion, motility, and invasion in cells that overexpress uPAR relative to vector control cells. Distinct alterations of focal adhesion protein expression and phosphorylation, including p130cas and paxillin were observed, suggestive of enhanced focal adhesion turnover. Immunohistochemical analysis of microarrayed human OSCC revealed a significant correlation between uPAR and p130cas expression. The non-receptor protein tyrosine kinase c-Src was responsible for the phosphorylation of p130cas in response to uPAR/alpha3beta1/laminin-5 engagement. Further downstream, the Rho family GTPase Cdc42, but not Rac1, was activated, suggesting a pathway leading to actin reorganization, filopodial protrusion and enhanced motility in uPAR overexpressing oral cancer cells. These data shed light on a molecular mechanism whereby acquisition of uPAR expression may modulate OSCC invasive activity through alteration of focal adhesion dynamics. PMID- 21630092 TI - Id1-induced inhibition of p53 facilitates endothelial cell migration and tube formation by regulating the expression of beta1-integrin. AB - The Id1 protein is critical for endothelial cell angiogenesis, and this function is particularly relevant to cancer development, cardiovascular disease, and wound healing. We hypothesized that Id1 enhanced migration and tubulogenesis by controlling the expression and function of p53. In this study, we examined cell migration following Id1 overexpression and silencing endothelial cells. The results showed that overexpression of Id1 enhanced cell migration and increased beta1-integrin expression, but inhibition of beta1-integrin blocked motility even in clones overexpressing Id1, suggesting that Id1 regulated motility through beta1-integrin. Further analysis revealed that p53, whose expression and distribution is regulated by Id1, was critical for cell migration, and may be involved in regulating the expression of beta1-integrin. Inhibiting p53 function using PFT-alpha, a functional inhibitor of p53, increased the expression of beta1 integrin and promoted cell migration even in Id1-silencing endothelial cells, demonstrating that the Id1 knockdowns induced inhibition of endothelial cell migration and the expression of beta1-integrin were controlled by p53. In addition, Id1-p53 pathway regulated the cytoskeleton formation and tubulogenesis. These results demonstrate that Id1-induced beta1-integrin expression in endothelial cells and the function of Id1 in cell migration and tubulogenesis are dependent on p53. PMID- 21630093 TI - Anticorrosion and cytocompatibility behavior of MAO/PLLA modified magnesium alloy WE42. AB - Recently, biodegradable magnesium alloys have been introduced in the field of cardiovascular stents to avoid the specific drawbacks of permanent metallic implants. However, the major obstacle of the clinical use of magnesium-based materials is their rapid corrosion rate. In this paper, a composite micro-arc oxidation/poly-L: -lactic acid (MAO/PLLA) coating was fabricated on the surface of the magnesium alloy WE42 to improve its corrosion resistance and the cytocompatibility of the modified materials was also investigated for safety aim. In our study, the morphology of materials was analyzed by Scanning electron microscopy. Potentiodynamic polarization was used to evaluate the corrosion behavior of the samples and corrosion weight loss was used to demonstrate their degradation rate. Furthermore, we applied cytotoxicity test in testing the cytocompatibility of the modified samples. The results showed that the PLLA coating effectively sealed the microcracks and micropores on the surface of the MAO coating by physical interlocking to interfere the corrosion ions. The corrosion rate was decreased and the cyototoxicity test showed that the MAO/PLLA composite coating WE42 had good cytocompatibility. PMID- 21630094 TI - Ultra-high field diffusion tensor imaging of articular cartilage correlated with histology and scanning electron microscopy. AB - OBJECT: To investigate the relationship of the different diffusion tensor imaging (DTI) parameters (ADC, FA, and first eigenvector (EV)) to the constituents (proteoglycans and collagen), the zonal arrangement of the collagen network, and mechanical loading of articular cartilage. MATERIAL AND METHODS: DTI of eight cartilage-on-bone samples of healthy human patellar cartilage was performed at 17.6 T. Three samples were additionally imaged under indentation loading. After DTI, samples underwent biomechanical testing, safranin-O staining for semiquantitative proteoglycan estimation, and scanning electron microscopy (SEM) for depicting collagen architecture. RESULTS: From the articular surface to the bone-cartilage interface, ADC continuously decreased and FA increased. Cartilage zonal heights calculated from EVs strongly correlated with SEM-derived zonal heights (P < 0.01, r (2)=0.87). Compression reduced ADC in the superficial 30% of cartilage and increased FA in the superficial 5% of cartilage. Reorientation of the EVs indicative of collagen fiber reorientation under the indenter was observed. No significant correlation was found between ADC, FA, and compressive stiffness. CONCLUSIONS: Correlating ADC and FA with proteoglycan and collagen content suggests that diffusion is dominated by different depth-dependent mechanisms within cartilage. Knowledge of the spatial distribution of the DTI parameters and their variation contributes to form a database for future analysis of defective cartilage. PMID- 21630098 TI - Genome-wide analysis of the RING finger gene family in apple. AB - The RING finger protein family plays a crucial role in plant growth and development and in response to biotic and abiotic stresses. However, no detailed information concerning this family is available for apple (Malus * domestica L. Borkh) due to the limited information on whole genome sequences. In this study, 688 RING domains in 663 predicted proteins were identified in apple. Based on the spacing between metal ligands or substitutions at one or more of the metal ligand positions, nine RING types were identified: RING-H2, RING-HC, RING-C2, RING-v, RING-D, RING-S/T, RING-G, RING-mH2, and RING-mHC, in which the first seven types were described previously in Arabidopsis, while the latter two were newly identified in apple. Proteins containing RING finger motifs were further classified into 57 groups according to the different known or unknown domains outside the RING domains. A total of 643 retrieved proteins appear to be distributed over all 17 linkage groups with different densities. Microarray and expressed sequence tag data revealed that only a few of these RING finger proteins may be involved in fruit development. As a first step towards genome wide analyses of the RING-containing genes in apple, our results provide valuable information for understanding the classification and putative functions of the RING finger gene family in higher plants. PMID- 21630099 TI - A novel gammaherpesvirus isolated from a black-tailed prairie dog (Cynomys ludovicianus). AB - A new gammaherpesvirus, tentatively named cynomys herpesvirus 1 (CynGHV-1), was isolated from a black-tailed prairie dog (Cynomys ludovicianus). CynGHV-1 replicated cytopathogenically to moderate titers in various cell lines. Ten kb of the CynGHV-1 genome was sequenced using degenerate PCR and genomic cloning. Sequence similarities were found to different genes from known gammaherpesviruses. Phylogenetic analysis suggested that CynGHV-1 was in fact a novel virus closely related to representatives of different genera and unclassified members of the subfamily Gammaherpesvirinae. However, CynGHV-1 could not be assigned to any particular genus and therefore remains unclassified. PMID- 21630100 TI - Bone-prognostic status after cessation of cadmium exposure for one month in male rats. AB - This study investigated bone status after decreased cadmium (Cd) exposure in male rats. Sprague-Dawley male rats were randomly divided into three groups. One group was injected subcutaneously with sodium chloride as control. The others were given CdCl2 by subcutaneous injection at doses of 0.5 mg Cd/kg body weight (bw) for 2 months (Cd+2m) and for 3 months (Cd+3m). For the Cd+2m group, the rats were shifted to cessation of Cd injection for 1 month after 2 months' exposure. At month 3, micro-computed tomography (micro-CT) analyses were performed on the proximal tibia and lumbar spine, and urine was collected from all rats. Rats were then killed and blood collected for metabolic-marker measurement and Cd assay. Bone tissues were also collected for bone-mass assay, biomechanical test, and bone-histology analysis. Cd burdens of rats in the Cd+2m and Cd+3m groups were both significantly greater than those in the control group. Cd burdens of rats were lower in the Cd+2m group compared with the Cd+3m group. Bone damage occurred in the Cd+2m and Cd+3m groups compared with the control group (p<0.05), but no significant improvement was found in the Cd+2m group compared with the Cd+3m group. Cd damage to bone could not be reversed over the short term. More attention should be paid to Cd's toxic effects on bone after decreased exposure. PMID- 21630101 TI - Effect of phototherapy on gastrointestinal smooth muscle activity and oxidative stress. AB - AIM: To evaluate the effect of phototherapy on gastrointestinal smooth muscle activity and oxidative stress. METHODS: Wistar albino rats (n = 18, in the first 7 days of life) weighing 7 +/- 2 g with both sexes were included in the study. The animals were randomized into three groups. In control group (CG), median laparotomy was performed to obtain 1 cm of jejunum, terminal ileum and colonic segments. In the phototherapy group (PTG), led phototherapy with a wave density of 40 MUw/cm(2)/nm were used (Bilitron 3006, Fanem, Brasil). The efficacy surface of phototherapy was 30-40 cm and the exposure distance was 30 cm. The duration of phototherapy was 24 h. Sham group (SG) received white light with the same wave density and exposure distance. The oxidative stress markers and contraction responses were investigated from intestinal segments obtained from experiments. RESULTS: The jejunum segments showed significantly lowered contraction response to carbachol in SG when compared to CG and PTG (p < 0.05). Decreased contractile response to KCl was detected in both SG and PTG in terminal ileum segments. MDA levels showed no difference between groups (p > 0.05). Total sulfhydryl (T-SH) levels were found significantly increased in PTG when compared to CG and SG (p < 0.05). When NO levels were evaluated, NO levels were found decreased in PTG and SG with respect to CG (p < 0.05). CONCLUSION: PT may cause various alterations in oxidant/antioxidant system in intestinal segments. Unlike to clinical findings, decreased contractile responses were detected in rat gastrointestinal smooth muscles after PT. PMID- 21630103 TI - Teaching uncommon and highly complex operations: maximizing the teaching and learning. AB - Teaching complex and uncommon operative procedures require instruction on technical skill and cognitive decision-making frameworks. To understand this dynamic process, several adult learning theories are discussed including the Fitts-Posner's three stage theory, transfer learning theory, and expert performance theory. By understanding how trainees can deconstruct complex task and transfer learning from more basic operative cases, surgical educators can reflect on how they structure and stratify operative and simulation experiences to maximize learning. PMID- 21630102 TI - MTHFD1 G1958A, BHMT G742A, TC2 C776G and TC2 A67G polymorphisms and head and neck squamous cell carcinoma risk. AB - Alterations in folate metabolism may contribute to the process of carcinogenesis by influencing DNA methylation and genomic stability. Polymorphisms in genes encoding enzymes involved in this pathway may alter enzyme activity and consequently interfere in concentrations of homocysteine and S-adenosylmethionine that are important for DNA synthesis and cellular methylation reactions. The objectives were to investigate MTHFD1 G1958A, BHMT G742A, TC2 C776G and TC2 A67G polymorphisms involved in folate metabolism on head and neck cancer risk and the association between these polymorphisms with risk factors. Polymorphisms were investigated in 762 individuals (272 patients and 490 controls) by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and Real Time PCR. Chi-square and Multiple logistic regression were used for the statistical analysis. Multiple logistic regression showed that tobacco and male gender were predictors for the disease (P < 0.05). Hardy-Weinberg equilibrium showed that the genotypic distributions were in equilibrium for both groups in all polymorphisms studied. The BHMT 742GA or AA genotypes associated with tobacco consumption (P = 0.016) increase the risk for head and neck squamous cell carcinoma (HNSCC). The present study suggests that BHMT 742GA polymorphism associated to tobacco modulate HNSCC risk. However, further investigation of gene-gene interactions in folate metabolism and studies in different populations are needed to investigate polymorphisms and HNSCC risk. PMID- 21630104 TI - Video: totally laparoscopic left lateral segmentectomy for hepatic malignancies: a modified technique. AB - We present our series of over 30 totally laparoscopic left hepatic lobectomies (hepatic segments II/III) performed only for malignancy. The short- and long-term results support this technique as safe and efficient. This video will illustrate the pertinent issues regarding trocar placement, intrahepatic anatomy, and the technical maneuvers necessary to perform the modified approach using totally laparoscopic techniques. PMID- 21630105 TI - Efficacy of an emergency department-based HIV screening program in the Deep South. AB - Human Immunodeficiency Virus (HIV) and Acquired Immunodeficiency Syndrome (AIDS) continue to be a significant public health concern in the United States. It disproportionately affects persons in the Deep South of the United States, specifically African Americans. This is a descriptive report of an Emergency Department (ED)-based HIV screening program in the Deep South using the 2006 Centers for Disease Control and Prevention (CDC) recommendations for rapid testing and opt-out consent. Between May 2008 and March 2010, patients presenting for medical care to the ED Monday through Friday between 10 AM: and 10 PM: were approached for HIV screening. Patients were eligible for screening if they were 18 or older, had no previous history of positive HIV tests, were English Speaking, and were not incarcerated, medically unstable, or otherwise able to decline testing. All patients were tested using the OraQuick(r) rapid HIV 1/2 antibody test. Patients with non-reactive results were referred to community anonymous testing sites for further testing. Patients with reactive results had confirmatory Western blot and CD4 counts drawn and were brought back to the ED for disclosure of the results. All patients with confirmed HIV positive via reactive Western blot were referred to the hospital-based infectious disease clinic or county health department. We tested 7,616 patients out of 8,922 approached. The overall test acceptance rate was 85.4%. 91.0% of patients tested were African American. The most common reason for refusal was recent HIV test. 1.7% of patients tested were confirmed HIV positive via Western blot. 95.2% of patients testing HIV positive were African American. The average CD4 count for patients testing positive was 276 cells/MUl, with 42.0% of patients having CD4 counts <=200 MUl, consistent with an AIDS diagnosis. 88.4% of patients who had reactive oral swabs returned for Western blot results and 75.0% of patients attended their first clinic visit. We have been able to successfully carry out an ED-based HIV screening program in a resource-poor urban teaching facility in the Deep South. We define our success based on our relatively high test acceptance rate and high rate of attendance at first clinic visit. Our patient population has a relatively high undocumented HIV prevalence and are at advanced stage of disease at the time of diagnosis. PMID- 21630106 TI - Fatal injuries in the slums of Nairobi and their risk factors: results from a matched case-control study. AB - Injuries contribute significantly to the rising morbidity and mortality attributable to non-communicable diseases in the developing world. Unfortunately, active injury surveillance is lacking in many developing countries, including Kenya. This study aims to describe and identify causes of and risk factors for fatal injuries in two slums in Nairobi city using a demographic surveillance system framework. The causes of death are determined using verbal autopsies. We used a nested case-control study design with all deaths from injuries between 2003 and 2005 as cases. Two controls were randomly selected from the non-injury deaths over the same period and individually matched to each case on age and sex. We used conditional logistic regression modeling to identity individual- and community-level factors associated with fatal injuries. Intentional injuries accounted for about 51% and unintentional injuries accounted for 49% of all injuries. Homicides accounted for 91% of intentional injuries and 47% of all injury-related deaths. Firearms (23%) and road traffic crashes (22%) were the leading single causes of deaths due to injuries. About 15% of injuries were due to substance intoxication, particularly alcohol, which in this community comes from illicit brews and is at times contaminated with methanol. Results suggest that in the pervasively unsafe and insecure environment that characterizes the urban slums, ethnicity, residence, and area level factors contribute significantly to the risk of injury-related mortality. PMID- 21630107 TI - A peptide epitope derived from the cancer testis antigen HOM-MEL-40/SSX2 capable of inducing CD4+ and CD8+ T-cell as well as B-cell responses. AB - BACKGROUND: Antigen-derived HLA class I-restricted peptides can generate specific CD8(+) T-cell responses in vivo and are therefore often used as vaccines for patients with cancer. However, only occasional objective clinical responses have been reported suggesting the necessity of CD4(+) T-cell help and possibly antibodies for the induction of an effective anti-tumor immunity in vivo. The SSX2 gene encodes the cancer testis antigen (CTA) HOM-MEL-40/SSX2, which is frequently expressed in a wide spectrum of cancers. Both humoral and cellular immune responses against SSX2 have been described making SSX2 an attractive candidate for vaccine trials. METHODS: SYFPEITHI algorithm was used to predict five pentadecamer peptides with a high binding probability for six selected HLA DRB1 subtypes (*0101, *0301, *0401, *0701, *1101, *1501) which are prevalent in the Caucasian population. RESULTS: Using peripheral blood cells of 13 cancer patients and 5 healthy controls, the HOM-MEL-40/SSX2-derived peptide p101-111 was identified as an epitope with dual immunogenicity for both CD4(+) helper and cytotoxic CD8(+) T cells. This epitope also reacted with anti-SSX2 antibodies in the serum of a patient with breast cancer. Most remarkably, SSX2/p101-111 simultaneously induced specific CD8, CD4, and antibody responses in vitro. CONCLUSIONS: p101-111 is the first CTA-derived peptide which induces CD4(+), CD8(+), and B-cell responses in vitro. This triple-immunogenic peptide represents an attractive vaccine candidate for the induction of effective anti-tumor immunity. PMID- 21630108 TI - Physical activity and fruit and vegetable intake among American Indians. AB - The American Indian population has among the highest rates of obesity in the United States. Thus, it is critical to understand factors related to this epidemic (e.g., physical activity, nutrition) among this ethnic minority population. The current study examined factors related to engaging in at least 4 days of physical activity (PA) per week and factors related to consuming at least 5 fruits and vegetables (FV) per day among a sample of American Indians in the Midwest. We used multiple methods to recruit participants for this study, including recruitment at pow wows, focus groups, health fairs, new student orientation for American Indian students, and other venues. A total of 998 American Indians (76% participation rate) completed a survey assessing sociodemographics, physical activity level, fruit and vegetable intake, and perceptions regarding the recommendations for physical activity and fruit and vegetable intake. Factors associated with exercising >=4 days in the past week (44.77% of the sample) include being younger (P = .002), being male (P < .001), having at least some college education (P = .048), eating >=5 FV per day, and higher perceived number of days of PA recommended (P < .001). Factors associated with eating >=5 servings of FV per day (37.01% of the sample) included exercising >=4 days in the past week (P < .001) and higher perceived number of servings of FV recommended (P < .001). These findings highlight the importance of education in enhancing engagement in positive weight control behaviors and the importance of addressing both physical activity and nutrition among the American Indian population. PMID- 21630109 TI - Venue staff knowledge of their patrons' gambling and problem gambling. AB - In several jurisdictions around the world, venue staff are encouraged to make reasonable attempts to determine whether patrons may be experiencing problems associated with their gambling. Although a number of visible indicators and behaviours are recognised as being indicative of gambling problems, no research has investigated how effective staff might be in identifying problem gamblers in venues. The aim of this field test was to examine the level of patron familiarity amongst staff working in small to moderate-sized venues. Patrons (n = 303) were asked to complete a short survey including the Problem Gambling Severity Index and venue staff were asked to describe their knowledge of the same patrons independently. The results showed that patrons rated as more at risk by staff scored significantly higher on the PGSI, but that point-in-time staff ratings were not sufficiently accurate to allow effective identification of problem gamblers. The importance of using accumulated information over multiple sessions as well as technological monitoring of behaviour was emphasised by these results. PMID- 21630110 TI - Established and emerging dose reduction methods in cardiac computed tomography. AB - Cardiac computed tomography (CT) is a non-invasive modality that is commonly used as an alternative to invasive coronary angiography for the investigation of coronary artery disease. The enthusiasm for this technology has been tempered by a growing appreciation of the potential risks of malignancy associated with the use of ionising radiation. In the spirit of minimizing patient risk, the medical profession and industry have worked hard to developed methods and protocols to reduce patient radiation exposure while maintaining excellent diagnostic accuracy. A complete understanding of radiation reduction techniques will allow clinicians to reduce patient risk while providing an important diagnostic service. This review will consider the established and emerging techniques that may be adopted to reduce patient absorbed doses from x-ray CT. By modifying (1) x ray tube output, (2) imaging time (scan duration), (3) imaging distance (scan length) and (4) the appropriate use of shielding, clinicians will be able to adhere to the 'as low as reasonably achievable (ALARA)' principle. PMID- 21630111 TI - Can we manage for resilience? The integration of resilience thinking into natural resource management in the United States. AB - The concept of resilience is now frequently invoked by natural resource agencies in the US. This reflects growing trends within ecology, conservation biology, and other disciplines acknowledging that social-ecological systems require management approaches recognizing their complexity. In this paper, we examine the concept of resilience and the manner in which some legal and regulatory frameworks governing federal natural resource agencies have difficulty accommodating it. We then use the U.S. Forest Service's employment of resilience as an illustration of the challenges ahead. PMID- 21630112 TI - Lateralization of frequency-specific networks for covert spatial attention to auditory stimuli. AB - We conducted a cued spatial attention experiment to investigate the time frequency structure of human EEG induced by attentional orientation of an observer in external auditory space. Seven subjects participated in a task in which attention was cued to one of two spatial locations at left and right. Subjects were instructed to report the speech stimulus at the cued location and to ignore a simultaneous speech stream originating from the uncued location. EEG was recorded from the onset of the directional cue through the offset of the inter-stimulus interval (ISI), during which attention was directed toward the cued location. Using a wavelet spectrum, each frequency band was then normalized by the mean level of power observed in the early part of the cue interval to obtain a measure of induced power related to the deployment of attention. Topographies of band specific induced power during the cue and inter-stimulus intervals showed peaks over symmetric bilateral scalp areas. We used a bootstrap analysis of a lateralization measure defined for symmetric groups of channels in each band to identify specific lateralization events throughout the ISI. Our results suggest that the deployment and maintenance of spatially oriented attention throughout a period of 1,100 ms is marked by distinct episodes of reliable hemispheric lateralization ipsilateral to the direction in which attention is oriented. An early theta lateralization was evident over posterior parietal electrodes and was sustained throughout the ISI. In the alpha and mu bands punctuated episodes of parietal power lateralization were observed roughly 500 ms after attentional deployment, consistent with previous studies of visual attention. In the beta band these episodes show similar patterns of lateralization over frontal motor areas. These results indicate that spatial attention involves similar mechanisms in the auditory and visual modalities. PMID- 21630113 TI - Effects of testing on learning of functions. AB - Is learning of a complex functional relationship enhanced by trying to predict what output will go with a given input, as compared to studying an input-output pair? We examined learning of a bilinear function and transfer to new items outside the trained range. Subjects either saw the input-output pairs (study-only condition) or attempted to guess the output and then saw the pair (test/study condition). The total study times were equated, and motivation was enhanced with a monetary bonus. Performance was markedly better for the test/study condition, both within the trained range and in the transfer test. This benefit of testing during training was observed on a criterial test administered shortly after training. Testing has long been shown to enhance the explicit learning and retention of verbal material; our present findings reveal a novel domain for which testing can also be advantageous-that is, function learning. PMID- 21630114 TI - On perfect working-memory performance with large numbers of items. AB - Many popular models conceptualize working memory as consisting of three or four discrete slots or bins. This conceptualization, however, has been seemingly refuted by Bays and Husain (2009), who reported perfect performance on a working memory task with a large number of very simple items. We show, however, that this perfect-performance result likely reflects a design flaw rather than mnemonic structure. The flaw is that the test array itself in Bays and Husain's study provides information about the correct answer without recourse to working memory. We show perfect performance on eight items for 18 participants when this information is present. We show that performance is poorer, however, when this information is removed. Hence, the Bays and Husain result does not threaten models that stipulate that working memory is composed of limited slots. PMID- 21630115 TI - Postsynaptic degeneration as revealed by PSD-95 reduction occurs after advanced Abeta and tau pathology in transgenic mouse models of Alzheimer's disease. AB - Impairment of synaptic plasticity underlies memory dysfunction in Alzheimer's disease (AD). Molecules involved in this plasticity such as PSD-95, a major postsynaptic scaffold protein at excitatory synapses, may play an important role in AD pathogenesis. We examined the distribution of PSD-95 in transgenic mice of amyloidopathy (5XFAD) and tauopathy (JNPL3) as well as in AD brains using double labeling immunofluorescence and confocal microscopy. In wild type control mice, PSD-95 primarily labeled neuropil with distinct distribution in hippocampal apical dendrites. In 3-month-old 5XFAD mice, PSD-95 distribution was similar to that of wild type mice despite significant Abeta deposition. However, in 6-month old 5XFAD mice, PSD-95 immunoreactivity in apical dendrites markedly decreased and prominent immunoreactivity was noted in neuronal soma in CA1 neurons. Similarly, PSD-95 immunoreactivity disappeared from apical dendrites and accumulated in neuronal soma in 14-month-old, but not in 3-month-old, JNPL3 mice. In AD brains, PSD-95 accumulated in Hirano bodies in hippocampal neurons. Our findings support the notion that either Abeta or tau can induce reduction of PSD 95 in excitatory synapses in hippocampus. Furthermore, this PSD-95 reduction is not an early event but occurs as the pathologies advance. Thus, the time dependent PSD-95 reduction from synapses and accumulation in neuronal soma in transgenic mice and Hirano bodies in AD may mark postsynaptic degeneration that underlies long-term functional deficits. PMID- 21630116 TI - Tracheal intubation during chest compressions using Pentax-AWS((r)), GlideScope ((r)), and Macintosh laryngoscope: a randomized crossover trial using a mannequin. AB - BACKGROUND: A randomized crossover trial was conducted to compare the performance of two videolaryngoscopes (Pentax-AWS((r)), GlideScope((r))) with the Macintosh laryngoscope for tracheal intubation during continuous chest compressions on a mannequin. METHODS: Thirty-two inexperienced junior interns performed tracheal intubations on an advanced life support simulator with either a normal or difficult airway scenario. The sequence of intubating devices and airway difficulty were randomized. The following data were measured and recorded: time to complete tracheal intubation (primary end point), overall success rate, time to visualize the vocal cords, percentage of glottic opening, dental compression, and ease of intubation. RESULTS: With a normal airway, the times (median [interquartile range]) to complete tracheal intubation were shorter with the Pentax-AWS (12.1 [10.1-14.4] sec) and the GlideScope (14.3 [12.4-17.6] sec) than with the Macintosh laryngoscope (16.5 [13.1-22.1] sec) (P < 0.03 for both). The time difference between the two videolaryngoscopes was not statistically significant. With a difficult airway scenario, the times to complete tracheal intubation were 13.9 [10.9-20.4] sec, 19.2 [16.4-32.3] sec, and 30.1 [21.0-56.5] sec, respectively (P < 0.05 for all differences). The videolaryngoscopes were also more effective than the Macintosh laryngoscope with respect to secondary outcomes. CONCLUSIONS: The two videolaryngoscopes were superior to the Macintosh laryngoscope in terms of performing tracheal intubation during continuous chest compressions on a mannequin. In a difficult airway scenario simulating cardiac arrest, the Pentax-AWS performed better than the GlideScope. PMID- 21630117 TI - Evolving challenges and opportunities for difficult airway management guidelines. PMID- 21630118 TI - Meta-analysis of desflurane and propofol average times and variability in times to extubation and following commands. AB - PURPOSE: We performed a meta-analysis to compare the operating room recovery time of desflurane with that of propofol. METHODS: Studies were included in which a) humans were assigned randomly to propofol or desflurane groups without other differences between groups (e.g., induction drugs) and b) mean and standard deviation were reported for extubation time and/or time to follow commands. Since there was heterogeneity of variance between treatment groups in the log-scale (i.e., unequal coefficients of variation of observations in the time scale), generalized pivotal methods for the lognormal distribution were used as inputs of the random effects meta-analyses. RESULTS: Desflurane reduced the variability (i.e., standard deviation) in time to extubation by 26% relative to propofol (95% confidence interval [CI], 6% to 42%; P = 0.006) and reduced the variability in time to follow commands by 39% (95% CI, 25% to 51%; P < 0.001). Desflurane reduced the mean time to extubation by 21% (95% CI, 9% to 32%; P = 0.001) and reduced the mean time to follow commands by 23% (95% CI, 16% to 30%; P < 0.001). CONCLUSIONS: The mean reduction in operating room recovery time for desflurane relative to propofol was comparable with that shown previously for desflurane relative to sevoflurane. The reduction in variability exceeded that of sevoflurane. Facilities can use the percentage differences when making evidence based pharmacoeconomic decisions. PMID- 21630119 TI - Anger regulation in traumatized Cambodian refugees: the perspectives of Buddhist monks. AB - Recent research has highlighted the importance of traditional methods of healing in relation to the treatment of psychological distress in non-Western populations. This pilot study, conducted in Massachusetts, investigates what Buddhist Cambodian monks consider to be the causes, phenomenology and appropriate intervention strategies for anger among Cambodian refugees. Six monks were interviewed at four major temples in Massachusetts. Findings suggested that anger was common in the Cambodian community, was frequently triggered by marital discord, and commonly resulted in verbal and physical violence and, sometimes, suicidality. Buddhist-based anger management strategies identified as useful by the monks included education about Buddhist doctrines, mindfulness meditation practices, and the use of herbal medication and holy water. These anger regulation strategies and treatments are discussed in the context of Buddhist beliefs and Western psychological interventions. PMID- 21630120 TI - No overt influence of lymphadenectomy on cancer-specific survival in organ confined versus locally advanced upper urinary tract urothelial carcinoma undergoing radical nephroureterectomy: a retrospective international, multi institutional study. AB - PURPOSE: Lymph node dissection (LND) is not routinely performed during radical nephroureterectomy (RNU) in upper tract urothelial carcinomas (UTUC), and its clinical relevance is unclear. The purpose of the present study was to evaluate the impact of LND on clinical outcomes in a large multicenter series of RNU for UTUC. METHODS: Detailed data on 785 patients subject to RNU were provided by nine international academic centers. The choice to perform lymphadenectomy was determined by the treating surgeon. All pathology slides were evaluated by dedicated genitourinary pathologists. Univariable and multivariable Cox regression models evaluated the association of nodal status with recurrence-free (RFS) and cancer-specific (CSS) survival. RESULTS: One hundred and ninety patients had LND. Pathological N stage was pN0 in 17%, pNx in 76%, and pN+ in 7%. The median follow-up period of the entire cohort was 34 months (interquartile range [IQR]: 15-65 months). Overall, five-year RFS and CSS estimates were 72.2 and 76%, respectively. In multivariable Cox regression analyses, pN0/pNx substaging was not an independent predictor of either RFS (hazard ratio [HR]: 1.1; P = 0.631) or CSS (HR: 1.3; P = 0.223). Similar results were obtained in a subgroup analysis limited to patients with organ-confined disease (HR: 0.9; P = 0.907 for RFS; HR: 0.4; P = 0.419 for CSS). Conversely, in patients with locally advanced disease, patients with pN0 disease have significantly better cancer related outcomes (HR: 0.3; P < 0.001 for RFS; HR: 0.3; P < 0.001 for CSS). CONCLUSION: The present series suggests pNx is more significantly associated with a worse prognosis than pN0, but only in patients with locally advanced UTUC. PMID- 21630121 TI - Erlotinib and pancreatic adenocarcinoma: uncertainty and hope. PMID- 21630122 TI - Needle versus excisional biopsy for noninvasive and invasive breast cancer: report from the National Cancer Data Base, 2003-2008. AB - BACKGROUND: Needle biopsy to diagnose breast cancer may soon become a quality measure for which hospitals are held accountable. This study examines the utilization of needle versus excisional biopsy in a contemporary cohort of patients and identifies factors associated with biopsy type. METHODS: Women with nonmetastatic, clinical Tis-T3 breast cancers diagnosed between 2003 and 2008 were selected from the National Cancer Data Base, which captures information from ~79% of breast cancers in the United States. Patients whose cancer was diagnosed by needle biopsy (fine-needle aspiration or core) were compared with patients diagnosed via excision, analyzing patient, hospital, and tumor characteristics. Logistic regression was used to identify important predictors of biopsy type. RESULTS: Of 373,837 patients, 303,677 (81.2%) underwent needle biopsy while 70,160 (18.8%) had excisional biopsy to diagnose their cancer. The needle biopsy rate increased from 73.8 to 86.7% whereas excisional biopsy declined from 26.2 to 13.3% over the study period (P < 0.001). In 2008, patients were statistically significantly more likely to undergo excisional biopsy if they had stage 0 disease; were treated at low-volume (<25 cases/year), community, or Atlantic census region hospitals; were <40 years old at diagnosis; were less educated; or were Asian/Pacific Islander (P < 0.001). The median rate of needle biopsy at high volume hospitals (>=140 cases/year) was 89.6%. CONCLUSION: The use of needle biopsy is increasing. Tumor stage, hospital volume, and hospital location were the most statistically significant predictors of biopsy type. Rates of needle biopsy at high-volume hospitals suggest that appropriate utilization of this preferred diagnostic method should approach 90%. PMID- 21630123 TI - The safety of multiple re-excisions after lumpectomy for breast cancer. AB - BACKGROUND: Breast cancer patients may undergo multiple re-excisions after lumpectomy in an attempt to obtain clear margins and avoid mastectomy. We sought to determine the overall local recurrence rate and surgical outcome of patients undergoing two or more re-excisions and to identify predictors of success in attaining clear margins. METHODS: Retrospective review of breast cancer patients who underwent lumpectomy for invasive cancer or ductal carcinoma in situ (DCIS) from 1997 to 2007. Patients who underwent two or more re-excisions were identified and analyzed. RESULTS: We identified 3,737 patients who underwent lumpectomy over this 10-year period. 875 (23.4%) had close or positive margins requiring a second procedure; 797 (91.1%) had a re-excision; and 78 (8.9%) went directly to mastectomy. Seventy patients underwent multiple re-excisions; 66 patients had 2 re-excisions, 3 patients had 3 re-excisions, and 1 patient had 4 re-excisions. 70% (49/70) of multiple re-excision patients achieved clear margins (26 DCIS, 35 T1, 8 T2, and 1 T3 tumors). All 49 patients who successfully treated with multiple re-excisions received radiation. At a median follow-up of 64 months, 1 of 49 (2.0%) patients had an in-breast recurrence, and 1 of 49 (2.0%) patients had a distant recurrence. Statistically significant risk factors for persistently involved margins after two re-excisions included multifocality and positive lymph node status. CONCLUSION: Multiple re-excisions to obtain clear margins are a safe alternative to mastectomy for women with invasive cancer or DCIS. There is an acceptably low risk of local and systemic failure when negative margins are ultimately achieved. PMID- 21630124 TI - Outcomes of screening-detected ductal carcinoma in situ treated with wide excision alone. AB - BACKGROUND: Ductal carcinoma in situ (DCIS) is commonly identified on screening mammography. Standard treatment for localized DCIS is wide local excision (WLE) and adjuvant radiotherapy. This approach represents overtreatment in many cases, where the DCIS would never have become clinically significant, or where less intensive treatment would have been satisfactory. We reviewed the medium-term outcome of a cohort of screen detected DCIS patients treated mainly with WLE without radiotherapy. METHODS: All patients diagnosed with DCIS at NorthWestern BreastScreen between January 1994 and December 2005 were identified from a prospective database. Demographic, pathological, treatment, and outcome data were collected and analyzed. Survival and local recurrence (LR) rates were determined, and associations between various factors and recurrence were analyzed. RESULTS: A total of 422 patients were diagnosed with DCIS. There were 400 patients treated with WLE, and 27 of these received adjuvant radiotherapy. The 5- and 8-year overall and breast cancer specific survival rates were 96.1 and 91.3%, and 99.6 and 99.3%, respectively. The local recurrence rate was 15.4 and 17.1% at 5 and 8 years. Of 56 local recurrences, 34 had WLE after recurrence, 16 of which had adjuvant radiotherapy. No single factor was statistically significantly associated with local recurrence, although combining factors revealed groups where the LR rate was less than 5%. CONCLUSIONS: Breast cancer specific mortality was very low in this cohort of older patients with screen-detected DCIS. There was a moderate rate of local recurrence that could usually be salvaged with breast conservation. Decisions regarding adjuvant radiotherapy should take these findings into account. PMID- 21630125 TI - High expression of Y-box-binding protein-1 is associated with poor survival in resectable esophageal squamous cell carcinoma. AB - BACKGROUND: Y-box-binding protein-1 (YB-1) is a multifunctional protein that regulates gene expression through both transcriptional and translational mechanism. Its expression has been associated with tumor progression and poor prognosis of many cancers. However, its role and clinical significance in resectable esophageal squamous cell carcinoma (ESCC) is still scanty. The purpose of this study was to investigate the prognostic significance of YB-1 expression by immunohistochemistry in a group of patients with ESCC treated with surgical resection. METHODS: Tissue microarray that included 233 surgically resected ESCC specimens and 49 cases of adjacent normal tissues was successfully generated for immunohistochemical evaluation. The clinical/prognostic significance of YB-1 expression was statistically analyzed. Kaplan-Meier analysis was used to compare the postoperative survival between groups. RESULTS: The results showed the immunostaining of YB-1 was distributed predominantly in cytoplasm in tumor cells, which occurred in all of the 233 patients. A higher recurrence (disease-free survival) and lower survival (overall survival) of ESCC was found in patients whose tissues had increased YB-1 expression (P<.001/P=.001). Furthermore, YB-1 expression could stratify the patient survival (disease-free survival/overall survival) in stage II (P=.012/.016). The Cox proportionate hazard regression model also established that high YB-1 expression was significantly correlated with increased risk (RR=1.752) of recurrence compared with lowYB-1 expression (P=.004). CONCLUSIONS: High expression of YB-1 is associated with poor survival in resectable ESCC patients. PMID- 21630126 TI - Incidence and progression of diabetic retinopathy in Japanese adults with type 2 diabetes: 8 year follow-up study of the Japan Diabetes Complications Study (JDCS). AB - AIMS/HYPOTHESIS: The aim of this study was to determine the incidence and progression rates of diabetic retinopathy and their associations in Japanese individuals with type 2 diabetes. METHODS: This is a part of the Japan Diabetic Complications Study (JDCS), a multi-centred randomised trial of type 2 diabetes patients aged 40-70 years with an 8 year follow-up. There were 1,221 patients without diabetic retinopathy at baseline; incidence of diabetic retinopathy was defined as the development of any diabetic retinopathy. There were 410 patients with mild non-proliferative diabetic retinopathy at baseline; progression of diabetic retinopathy was defined as the development of severe non-proliferative diabetic retinopathy or proliferative diabetic retinopathy. We used multivariate proportional Cox hazard models, and generalised additive models were also applied to identify potential threshold effect. RESULTS: The incidence and progression rate of diabetic retinopathy was 38.3/1,000 person-years and 21.1/1,000 person years, respectively. Higher HbA(1c) (adjusted HR [aHR] per 1% [10.9 mmol/mol] 1.36 [95% CI 1.28-1.45]), longer duration of diabetes (aHR per 5 year period 1.26 [95% CI 1.17-1.35]), higher systolic blood pressure (aHR per +10 mmHg 1.01 [95% CI 1.00-1.02]) and higher body mass index (aHR per 1 kg/m(2) 1.05 [95% CI 1.00 1.09]) were associated with incident diabetic retinopathy. The association between HbA(1c) and incident diabetic retinopathy was linear; the association with duration of diabetes increased rapidly between 5 and 10 years. Higher HbA(1c) was also associated with progression of diabetic retinopathy (aHR per 1% [10.9 mmol/mol] 1.66 [95% CI 1.41-1.96]). CONCLUSIONS: Observed incidence and progression rates of diabetic retinopathy seemed lower than that in western populations. HbA(1c) was the only factor associated with both incidence and progression of diabetic retinopathy. The strength of the association between duration of diabetes and incidence of diabetic retinopathy increased rapidly during a period of 5 to 10 years duration of diabetes. TRIAL REGISTRATION: C000000222 ( www.umin.ac.jp ) FUNDING: This study is supported by the Ministry of Health, Labour and Welfare, Japan. PMID- 21630127 TI - AAPS workshop report: strategies to address therapeutic protein-drug interactions during clinical development. AB - Therapeutic proteins (TPs) are increasingly combined with small molecules and/or with other TPs. However preclinical tools and in vitro test systems for assessing drug interaction potential of TPs such as monoclonal antibodies, cytokines and cytokine modulators are limited. Published data suggests that clinically relevant TP-drug interactions (TP-DI) are likely from overlap in mechanisms of action, alteration in target and/or drug-disease interaction. Clinical drug interaction studies are not routinely conducted for TPs because of the logistical constraints in study design to address pharmacokinetic (PK)- and pharmacodynamic (PD)-based interactions. Different pharmaceutical companies have developed their respective question- and/or risk-based approaches for TP-DI based on the TP mechanism of action as well as patient population. During the workshop both company strategies and regulatory perspectives were discussed in depth using case studies; knowledge gaps and best practices were subsequently identified and discussed. Understanding the functional role of target, target expression and their downstream consequences were identified as important for assessing the potential for a TP DI. Therefore, a question-and/or risk-based approach based upon the mechanism of action and patient population was proposed as a reasonable TP-DI strategy. This field continues to evolve as companies generate additional preclinical and clinical data to improve their understanding of possible mechanisms for drug interactions. Regulatory agencies are in the process of updating their recommendations to sponsors regarding the conduct of in vitro and in vivo interaction studies for new drug applications (NDAs) and biologics license applications (BLAs). PMID- 21630128 TI - Does training learners on simulators benefit real patients? AB - Despite limited data on patient outcomes, simulation training has already been adopted and embraced by a large number of medical schools. Yet widespread acceptance of simulation should not relieve us of the duty to demonstrate if, and under which circumstances, training learners on simulation benefits real patients. Here we review the data on performance of healthcare providers or trainees following simulation training, and discuss ways of enhancing transfer of learning from simulated to real patients. While there is tremendous potential for simulation in medical education and healthcare, further studies are needed to identify if and when simulation training improves the quality of care delivered to patients, and to compare the cost-effectiveness of simulated learning experiences to lower fidelity and less expensive interventions. PMID- 21630129 TI - Emerging trends in research on spatial and temporal organization of terpenoid indole alkaloid pathway in Catharanthus roseus: a literature update. AB - Catharanthus roseus (The Madagaskar Periwinkle) plant is commercially valued for harbouring more than 130 bioactive terpenoid indole alkaloids (TIAs). Amongst these, two of the leaf-derived bisindole alkaloids-vinblastine and vincristine are widely used in several anticancer chemotherapies. The great pharmacological values, low in planta occurrence, unavailability of synthetic substitutes and exorbitant market cost of these alkaloids have prompted scientists to understand the basic architecture and regulation of biosynthesis of these TIAs in C. roseus plant and its cultured tissues. The knowledge gathered over a period of 30 years suggests that the TIA biosynthesis is highly regulated by developmental and environmental factors and operates through a complex multi-step enzymatic network. Extensive spatial and temporal cross talking also occurs at inter- and intracellular levels in different plant organs during TIA biogenesis. A close association of indole, methylerythritol phosphate and secoiridoid monoterpenoid pathways and involvement of at least four cell types (epidermis, internal phloem associated parenchyma, laticifers and idioblasts) and five intracellular compartments (chloroplast, vacuole, nucleus, endoplasmic reticulum and cytosol) have been implicated with this biosynthetic mechanism. Accordingly, the research in this area is primarily advancing today to address and resolve six major issues namely: precise localization and expression of pathway enzymes using modern in situ RNA hybridization tools, mechanisms of intra- and intercellular trafficking of pathway intermediates, cloning and functional validation of genes coding for known or hitherto unknown pathway enzymes, mechanism of global regulation of the pathway by transcription factors, control of relative diversion of metabolite flux at crucial branch points and finally, strategising the metabolic engineering approaches to improve the productivity of the desired TIAs in plant or corresponding cultured tissues. The present literature update has been compiled to provide a brief overview of some of the emerging developments in our current understanding of TIA metabolism in C. roseus. PMID- 21630130 TI - Clinical practice guidelines for the prevention and treatment of EGFR inhibitor associated dermatologic toxicities. AB - BACKGROUND: Epidermal growth factor receptor inhibitors (EGFRI) produce various dermatologic side effects in the majority of patients, and guidelines are crucial for the prevention and treatment of these untoward events. The purpose of this panel was to develop evidence-based recommendations for EGFRI-associated dermatologic toxicities. METHODS: A multinational, interdisciplinary panel of experts in supportive care in cancer reviewed pertinent studies using established criteria in order to develop first-generation recommendations for EGFRI associated dermatologic toxicities. RESULTS: Prophylactic and reactive recommendations for papulopustular (acneiform) rash, hair changes, radiation dermatitis, pruritus, mucositis, xerosis/fissures, and paronychia are presented, as well as general dermatologic recommendations when possible. CONCLUSION: Prevention and management of EGFRI-related dermatologic toxicities is critical to maintain patients' health-related quality of life and dose intensity of antineoplastic regimens. More rigorous investigation of these toxicities is warranted to improve preventive and treatment strategies. PMID- 21630131 TI - CCN-2 is up-regulated by and mediates effects of matrix bound advanced glycated end-products in human renal mesangial cells. AB - CCN-2, also known as connective tissue growth factor (CCN-2/CTGF) is a cysteine rich, extracellular matrix protein that acts as a pro-fibrotic cytokine in tissues in many diseases, including in diabetic nephropathy. We have published that soluble advanced glycation end products (AGEs), that are present in increased amounts in diabetes, induce CCN-2. However in vivo AGEs are known to be heavily tissue bound and whether matrix bound AGEs regulate CCN-2 has not been investigated. In this study we determined in human renal mesangial cells if CCN-2 is induced by matrix associated AGEs and if CCN-2 may then secondarily mediate effects of matrix AGEs on extracellular matrix expansion. Data generated show that CCN-2 mRNA and protein expression are induced by matrix bound AGEs, and in contrast, this was not the case for TGF-beta1 mRNA regulation. Using CCN-2 adenoviral anti-sense it was found that CCN-2 mediated the up-regulation of fibronectin and the tissue inhibitor of matrix metalloproteinase, TIMP-1, that was caused by matrix bound AGEs. In conclusion, CCN-2 is induced by non enzymatically glycated matrix and it mediates downstream fibronectin and TIMP-1 increases, thus through this mechanism potentially contributing to ECM accumulation in the renal glomerulus in diabetes. PMID- 21630133 TI - First time proof of sage's tolerability and efficacy in menopausal women with hot flushes. AB - BACKGROUND: This trial aimed to assess the tolerability and efficacy of a fresh sage preparation in treating hot flushes and other menopausal complaints. Sage (Salvia officinalis) has been traditionally used to treat sweating and menopausal hot flushes, as well as to alleviate associated menopausal symptoms and as a general tonic. However, no clinical studies substantiating the use of sage in menopause have been published previously. METHODS: In an open, multicenter clinical trial conducted in eight practices in Switzerland, 71 patients (intent to-treat population [ITT], n=69; with a mean age of 56.4+/-4.7 years, menopausal for at least 12 months, and with at least five flushes daily) were recruited and treated with a once-daily tablet of fresh sage leaves for 8 weeks after an introductory baseline week. Parameters for the evaluation of efficacy were the change in intensity and frequency of hot flushes, and total score of the mean number of intensity-rated hot flushes (TSIRHF) as determined by diary protocol over the 2-month treatment period. Other variables included assessment of the Menopause Rating Scale (MRS) by the treating physician at baseline and after 2 months of therapy. RESULTS: In the ITT population there was a significant decrease in the TSIRHF by 50% within 4 weeks and by 64% within 8 weeks (P<0.0001). The mean total number of hot flushes per day decreased significantly each week from week 1 to 8. The mean number of mild, moderate, severe, and very severe flushes decreased by 46%, 62%, 79%, and 100% over 8 weeks, respectively. The MRS and its somato-vegetative, psychological, and urogenital subscales decreased significantly by 43%, 43%, 47%, and 20% respectively. The treatment was very well tolerated. CONCLUSION: A fresh sage preparation demonstrated clinical value in the treatment of hot flushes and associated menopausal symptoms. PMID- 21630132 TI - Cerebral gene expression and neurobehavioural development after perinatal exposure to an environmentally relevant polybrominated diphenylether (BDE47). AB - Nutrients in seafood are known to be beneficial for brain development. Effects of maternal exposure to 2,2',4,4' tetrabromo diphenylether (BDE47) was investigated, alongside the potential ameliorating impact of seafood nutrients, through assessment of neurobehaviour and gene expression in brain and liver. Developing mice were exposed during gestation and lactation via dams dosed through casein- or salmon-based feed, spiked with BDE47. Two concentrations were used: a low level (6 MUg/kg feed) representing an environmentally realistic concentration and a high level (1,900 MUg/kg feed) representing a BDE47 intake much higher than expected from frequents consumption of contaminated seafood. Experimental groups were similar with respect to reproductive success, growth and physical development. Minor, transient changes in neurobehavioural metrics were observed in groups given the highest dose of BDE47. No significant differences in behaviour or development were seen on postnatal day 18 among maternally exposed offspring. Cerebral gene expression investigated by microarray analyses and validated by RT-qPCR showed low fold changes for all genes, despite dose dependent accumulation of BDE47 in brain tissue. The gene for glutamate ammonia ligase was upregulated compared to control in the casein-based high BDE47diet, suggesting potential impacts on downstream synaptic transmission. The study supported a previously observed regulation of Igfbp2 in brain with BDE47 exposure. Genes for hepatic metabolic enzymes were not influenced by BDE47. Potential neurotoxic effects and neurobehavioural aberrations after perinatal exposure to high levels of BDE47 were not readily observed in mice pups with the present experimental exposure regimes and methods of analysis. PMID- 21630134 TI - Assisting oxidative protein folding: how do protein disulphide-isomerases couple conformational and chemical processes in protein folding? AB - Oxidative folding is the simultaneous process of forming disulphide bonds and native structure in proteins. Pathways of oxidative folding are highly diverse and in eukaryotes are catalysed by protein disulphide isomerases (PDIs). PDI consists of four thioredoxin-like domains, two of which contain active sites responsible for disulphide interchange reactions. The four domains are arranged in a horseshoe shape with the two active sites facing each other at the opening of the horseshoe. An extended hydrophobic surface at the bottom of the horseshoe is responsible for non-covalent, hydrophobic interactions with the folding protein. This binding site is capable of distinguishing between fully-folded and partially- or un-folded proteins. PDI is not only a catalyst of the formation of disulphide bonds, but also catalyses folding steps which involve significant conformational change in the folding protein. This review brings together the latest catalytic and structural data aimed at understanding how this is achieved. PMID- 21630135 TI - Multiscale approaches and perspectives to modeling aqueous electrolytes and polyelectrolytes. AB - We review recent work on scale-bridging modeling approaches applied to aqueous electrolytes and polyelectrolytes, connecting the local quantum chemical details to classical statistical and thermodynamics properties. We discuss solvation and pairing of ions in water, ways to include solvent degrees of freedom in effective ion-ion interactions, and coarse-grained simulations of polyelectrolytes including dielectric boundary effects. PMID- 21630136 TI - Prion protein and its conformational conversion: a structural perspective. AB - The key molecular event in the pathogenesis of prion diseases is the conformational conversion of a cellular prion protein, PrP(C), into a misfolded form, PrP(Sc). In contrast to PrP(C) that is monomeric and alpha-helical, PrP(Sc) is oligomeric in nature and rich in beta-sheet structure. According to the "protein-only" model, PrP(Sc) itself represents the infectious prion agent responsible for transmissibility of prion disorders. While this model is supported by rapidly growing experimental data, detailed mechanistic and structural aspects of prion protein conversion remain enigmatic. In this chapter we describe recent advances in understanding biophysical and biochemical aspects of prion diseases, with a special focus on structural underpinnings of prion protein conversion, the structural basis of prion strains, and generation of prion infectivity in vitro from bacterially-expressed recombinant PrP. PMID- 21630137 TI - Solid-state NMR techniques for the structural determination of amyloid fibrils. AB - This review discusses the solid-state NMR techniques developed for the study of amyloid fibrils. Literature up to the end of 2010 has been surveyed and the materials are organized according to five categories, viz. homonuclear dipolar recoupling and polarization transfer via J-coupling, heteronuclear dipolar recoupling, correlation spectroscopy, recoupling of chemical shift anisotropy, and tensor correlation. Our emphasis is on the NMR techniques and their practical aspects. The biological implications of the results obtained for amyloid fibrils are only briefly discussed. Our main objective is to showcase the power of NMR in the study of biological unoriented solids. PMID- 21630138 TI - Experimental systems to explore life origin: perspectives for understanding primitive mechanisms of cell division. AB - Compartmentalization is a necessary element for the development of any cell cycle and the origin of speciation. Changes in shape and size of compartments might have been the first manifestation of development of so-called cell cycles. Cell growth and division, processes guided by biological reactions in modern cells, might have originated as purely physicochemical processes. Modern cells use enzymes to initiate and control all stages of cell cycle. Protocells, in the absence of advanced enzymatic machinery, might have needed to rely on physical properties of the membrane. As the division processes could not have been controlled by the cell's metabolism, the first protocells probably did not undergo regular cell cycles as we know it in cells of today. More likely, the division of protocells was triggered either by some inorganic catalyzing factor, such as porous surface, or protocells divided when the encapsulated contents reached some critical concentration. PMID- 21630139 TI - Evolution of bet-hedging mechanisms in cell cycle and embryo development stimulated by weak linkage of stochastic processes. AB - Our current understanding of the origin and evolution of the cell cycle is largely filled with gaps and unresolved questions. Numerous similarities between the processes comprising the cell cycle in distant organisms from the Pro- and Eukaryota kingdoms provide some clues about the course that evolution has taken. Contemporary Prokaryotes and Eukaryotes regulate their cell cycles in a quite similar way, using a master oscillator that regulates cell division. Despite this striking similarity, they use entirely different molecules for this purpose. The necessity to keep the master oscillator intact for the survival of every cell/organism allows evolutionary changes in only the secondary mechanisms and processes of the cell cycle. This is especially clear in oocytes and embryos, which have a direct impact on the reproductive success of an adult organism. Here, we present examples of cues driving such mild evolutionary changes of certain aspects of cell cycle progression in oocytes and early embryos. We suggest that weak linkages between core processes that rely on randomness (stochasticity) have led to the evolution of strategies increasing fitness similar to bet-hedging, a stochastic-based survival strategy of risk minimization widely implemented by populations of bacteria, yeast, arthropods, and birds. Stochastic diversification of phenotypes by isogenic cells increases their fitness in unpredictable environments and improves their survival rate upon exposure to stress, a trait beneficial in evading antibiotic treatment by bacteria or withstanding chemotherapy by cancer cells. The evolution of bet hedging has been observed experimentally for bacteria and attributed to specific molecular mechanisms involved in this strategy. In this chapter, we set out to answer whether similar strategies could have evolved at the level of oocytes and embryos. We indicate possible evolutionary cues capable of realizing bet-hedging like mechanisms. PMID- 21630140 TI - Mechanics and regulation of cell shape during the cell cycle. AB - Many cell types undergo dramatic changes in shape throughout the cell cycle. For individual cells, a tight control of cell shape is crucial during cell division, but also in interphase, for example during cell migration. Moreover, cell cycle related cell shape changes have been shown to be important for tissue morphogenesis in a number of developmental contexts. Cell shape is the physical result of cellular mechanical properties and of the forces exerted on the cell. An understanding of the causes and repercussions of cell shape changes thus requires knowledge of both the molecular regulation of cellular mechanics and how specific changes in cell mechanics in turn effect global shape changes. In this chapter, we provide an overview of the current knowledge on the control of cell morphology, both in terms of general cell mechanics and specifically during the cell cycle. PMID- 21630141 TI - The spindle assembly checkpoint: clock or domino? AB - In each cell division, the newly duplicated chromosomes must be evenly distributed between the sister cells. Errors in this process during meiosis or mitosis are equally fatal: improper segregation of the chromosome 21 during human meiosis leads to Down syndrome (Conley, Aneuploidy: etiology and mechanisms, pp 35-89, 1985), whereas in somatic cells, aneuploidy has been linked to carcinogenesis, by unbalancing the ratio of oncogenes and tumor suppressors (Holland and Cleveland, Nat Rev Mol Cell Biol 10(7):478-487, 2009; Yuen et al., Curr Opin Cell Biol 17(6):576-582, 2005). Eukaryotic cells have developed a mechanism, known as the spindle assembly checkpoint, to detect erroneous attachment of chromosomes to the mitotic/meiotic spindle and delay the cell cycle to give enough time to resolve these defects. Research in the last 20 years, has demonstrated that the spindle assembly checkpoint is not only a pure checkpoint pathway, but plays a constitutive role in every cell cycle. Here, we review our current knowledge of how the spindle assembly checkpoint is integrated into the cell cycle machinery, and discuss some of the questions that have to be addressed in the future. PMID- 21630142 TI - Cell-size-dependent control of organelle sizes during development. AB - During development, cells differentiate into diverse cell types with different sizes. The size of intracellular organelles often correlates with the size of the cell, which may be important for cell homeostasis. The nucleus is a well-known example of an organelle whose size correlates with cell size. However, the mechanical basis of the correlation is unknown. The lengths of the mitotic spindle and contractile ring are emerging as model system to investigate the cell size-dependent control mechanisms of organelle size. Mechanistic models are proposed for the cell-size-dependent control of these organelles. Understanding the cell-size dependency of organelle sizes is expected to impact not only on the morphogenesis of the individual organelle, but also on cell homeostasis, cell cycle progression, and cell differentiation. PMID- 21630143 TI - The first cell cycle of the Caenorhabditis elegans embryo: spatial and temporal control of an asymmetric cell division. AB - Throughout the development of an organism, it is essential that the cell cycle machinery is fine-tuned to generate cells of different fate. A series of asymmetric cell divisions leads to lineage specification. The Caenorhabditis elegans embryo is an excellent system to study various aspects of the early embryonic cell cycle. The invariant nature of the rapid cell divisions is the key feature for studying the effects of small perturbations to a complex process such as the cell cycle. The thorough characterization of the asymmetric first cell division of the C. elegans embryo has given great insight on how the oscillations of the cell cycle coordinate with the cytoplasmic rearrangements that ultimately lead to two developmentally distinct daughter cells. PMID- 21630145 TI - Cell cycle in ascidian eggs and embryos. AB - In ascidians the cell cycle machinery has been studied mainly in oocytes while ascidian embryos have been used to dissect the mechanism that controls asymmetric cell division (ACD). Here we overview the most specific and often exceptional points and events in cell cycle control in ascidian oocytes and early embryos. Mature stage IV eggs are arrested at metaphase I due to cytostatic factor (CSF). In vertebrates, unfertilized eggs are arrested at metaphase II by CSF. Meta II CSF is mediated by the Mos/MEK/MAPK/Erp1 pathway, which inhibits the ubiquitin ligase APC/C(cdc20) preventing cyclin B destruction thus stabilizing MPF activity. CSF is inactivated by the fertilization Ca(2+) transient that stimulates the destruction of Erp1 thus releasing APC/C(cdc20) from inhibition. Although many of the components of CSF are conserved between the ascidian and the vertebrates, the lack of Erp1 in the ascidians (and indeed other invertebrates) is notable since the Mos/MAPK pathway nonetheless mediates Meta I-CSF. Moreover, since the fertilization Ca(2+) transient targets Erp1, it is not clear how the sperm-triggered Ca(2+) transient in ascidians (and again other invertebrates) stimulates cyclin B destruction in the absence of Erp1. Nonetheless, like mammalian eggs, sperm trigger a series of Ca(2+) oscillations that increases the rate of cyclin B destruction and the subsequent loss of MAPK activity leading to meiotic exit in ascidians. Positive feedback from MPF maintains the Ca(2+) oscillations in fertilized ascidian eggs ensuring the eventual loss of MPF stimulating the egg-to-embryo transition. Embryonic cell cycles in the ascidian are highly stereotyped where both the rate of cell division and the orientation of cell division planes are precisely controlled. Three successive rounds of ACD generate two small posterior germ cell precursors at the 64 cell stage. The centrosome-attracting body (CAB) is a macroscopic cortical structure visible by light microscopy that causes these three rounds of ACD. Entry into mitosis activates the CAB causing the whole mitotic spindle to rotate and migrate toward the cortical CAB leading to a highly ACD whereby one small cell is formed that inherits the CAB and approximately 40 maternal postplasmic/PEM RNAs including the germ cell marker vasa. PMID- 21630144 TI - EGG molecules couple the oocyte-to-embryo transition with cell cycle progression. AB - The oocyte-to-embryo transition is a precisely coordinated process in which an oocyte becomes fertilized and transitions to an embryonic program of events. The molecules involved in this process have not been well studied. Recently, a group of interacting molecules in C. elegans have been described as coordinating the oocyte-to-embryo transition with the advancement of the cell cycle. Genes egg-3, egg-4, and egg-5 represent a small class of regulatory molecules known as protein tyrosine phosphase-like proteins, which can bind phosphorylated substrates and act as scaffolding molecules or inhibitors. These genes are responsible for coupling the movements and activities of regulatory kinase mbk-2 with advancement of the cell cycle during the oocyte-to-embryo transition. PMID- 21630146 TI - Regulatory pathways coordinating cell cycle progression in early Xenopus development. AB - The African clawed frog, Xenopus laevis, is used extensively as a model organism for studying both cell development and cell cycle regulation. For over 20 years now, this model organism has contributed to answering fundamental questions concerning the mechanisms that underlie cell cycle transitions--the cellular components that synthesize, modify, repair, and degrade nucleic acids and proteins, the signaling pathways that allow cells to communicate, and the regulatory pathways that lead to selective expression of subsets of genes. In addition, the remarkable simplicity of the Xenopus early cell cycle allows for tractable manipulation and dissection of the basic components driving each transition. In this organism, early cell divisions are characterized by rapid cycles alternating phases of DNA synthesis and division. The post-blastula stages incorporate gap phases, lengthening progression, and allowing more time for DNA repair. Various cyclin/Cdk complexes are differentially expressed during the early cycles with orderly progression being driven by both the combined action of cyclin synthesis and degradation and the appropriate selection of specific substrates by their Cdk components. Like other multicellular organisms, chief developmental events in early Xenopus embryogenesis coincide with profound remodeling of the cell cycle, suggesting that cell proliferation and differentiation events are linked and coordinated through crosstalk mechanisms acting on signaling pathways involving the expression of cell cycle control genes. PMID- 21630147 TI - Control of DNA replication by cyclin-dependent kinases in development. AB - Cyclin-dependent kinases (CDKs) are required for initiation of DNA replication in all eukaryotes, and appear to act at multiple levels to control replication origin firing, depending on the cell type and stage of development. In early development of many animals, both invertebrate and vertebrate, rapid cell cycling is coupled with transcriptional repression, and replication initiates at closely spaced replication origins with little or no sequence specificity. This organisation of DNA replication is modified during development as cell proliferation becomes more controlled and defined. In all eukaryotic cells, CDKs promote conversion of "licensed" pre-replication complexes (pre-RC) to active initiation complexes. In certain circumstances, CDKs may also control pre-RC formation, transcription of replication factor genes, chromatin remodelling, origin spacing, and organisation of replication origin clusters and replication foci within the nucleus. Although CDK1 and CDK2 have overlapping roles, there is a limit to their functional redundancy. Here, I review these findings and their implications for development. PMID- 21630148 TI - Greatwall kinase, ARPP-19 and protein phosphatase 2A: shifting the mitosis paradigm. AB - Control of entry into mitosis has long been seen in terms of an explosive activation of cyclin-dependent kinase 1, the mitotic driver ensuring the phosphorylation of hundreds of proteins required for cell division. However, if these phosphorylations are maintained during M-phase, they must be removed when cells exit mitosis. It has been surmised that an "antimitotic" phosphatase must be inhibited to allow mitosis entry and activated for returning to interphase. This chapter discusses a series of recent works conducted on Xenopus egg extracts that provide the answers regarding the identity and the regulation of such a phosphatase. PP2A-B55delta is the major phosphatase controlling exit from mitosis; it is negatively regulated by the kinase Greatwall that phosphorylates the small protein ARPP-19 and converts it into a potent PP2A inhibitor. These findings provide a new element of paramount importance in the control of mitosis. PMID- 21630149 TI - The role of RanGTP gradient in vertebrate oocyte maturation. AB - The maturation of vertebrate oocyte into haploid gamete, the egg, consists of two specialized asymmetric cell divisions with no intervening S-phase. Ran GTPase has an essential role in relaying the active role of chromosomes in their own segregation by the meiotic process. In addition to its conserved role as a key regulator of macromolecular transport between nucleus and cytoplasm, Ran has important functions during cell division, including in mitotic spindle assembly and in the assembly of nuclear envelope at the exit from mitosis. The cellular functions of Ran are mediated by RanGTP interactions with nuclear transport receptors (NTRs) related to importin beta and depend on the existence of chromosome-centered RanGTP gradient. Live imaging with FRET biosensors indeed revealed the existence of RanGTP gradient throughout mouse oocyte maturation. NTR dependent transport of cell cycle regulators including cyclin B1, Wee2, and Cdc25B between the oocyte cytoplasm and germinal vesicle (GV) is required for normal resumption of meiosis. After GVBD in mouse oocytes, RanGTP gradient is required for timely meiosis I (MI) spindle assembly and provides long-range signal directing egg cortex differentiation. However, RanGTP gradient is not required for MI spindle migration and may be dispensable for MI spindle function in chromosome segregation. In contrast, MII spindle assembly and function in maturing mouse and Xenopus laevis eggs depend on RanGTP gradient, similar to X. laevis MII-derived egg extracts. PMID- 21630150 TI - Cell cycle control of germ cell differentiation. AB - The germ cell lineage is our lifelong reservoir of reproductive stem cells and our mechanism for transmitting genes to future generations. These highly specialised cells are specified early during development and then migrate to the embryonic gonads where sex differentiation occurs. Germ cell sex differentiation is directed by the somatic gonadal environment and is characterised by two distinct cell cycle states that are maintained until after birth. In the mouse, XY germ cells in a testis cease mitotic proliferation and enter G(1)/G(0) arrest from 12.5 dpc, while XX germ cells in an ovary enter prophase I of meiosis from 13.5 dpc. This chapter discusses the factors known to control proliferation and survival of germ cells during their journey of specification to sex differentiation during development. PMID- 21630151 TI - Protein kinases and protein phosphatases that regulate meiotic maturation in mouse oocytes. AB - Oocytes arrest at prophase of meiosis I (MI) and in vivo do not resume meiosis until they receive ovulatory cues. Meiotic resumption entails two rounds of chromosome segregation without an intervening round of DNA replication and an arrest at metaphase of meiosis II (MII); fertilization triggers exit from MII and entry into interphase. During meiotic resumption, there is a burst of protein phosphorylation and dephosphorylation that dramatically changes during the course of oocyte meiotic maturation. Many of these phosphorylation and dephosphorylation events are key to regulating meiotic cell cycle arrest and/or progression, chromosome dynamics, and meiotic spindle assembly and disassembly. This review, which is subdivided into sections based upon meiotic cell cycle stages, focuses on the major protein kinases and phosphatases that have defined requirements during meiosis in mouse oocytes and, when possible, connects these regulatory pathways. PMID- 21630152 TI - Anaphase-promoting complex control in female mouse meiosis. AB - Entry into, and passage through, the two meiotic divisions of the oocyte has to be highly coordinated to ensure proper segregation of chromosomes. This coordination ensures that the hallmark stops and starts of the meiotic process occur at the right time to prevent aneuploidy. The Anaphase-Promoting Complex is an activity mostly studied in the mitotic cell cycle division, where it has essential functions during mitosis. As detailed here the Anaphase-Promoting Complex also plays vital roles in controlling at least three meiotic events: maintenance of prophase I arrest, timely and faithful segregation of homologous chromosomes in meiosis I, and the meiotic arrest following ovulation. PMID- 21630153 TI - Established and novel Cdk/cyclin complexes regulating the cell cycle and development. AB - The identification of new members in the Cdk and cyclin families, functions for many of which are still emerging, has added new facets to the cell cycle regulatory network. With roles extending beyond the classical regulation of cell cycle progression, these new players are involved in diverse processes such as transcription, neuronal function, and ion transport. Members closely related to Cdks and cyclins such as the Speedy/RINGO proteins offer fresh insights and hope for filling in the missing gaps in our understanding of cell division. This chapter will present a broad outlook on the cell cycle and its key regulators with special emphasis on the less-studied members and their emerging roles. PMID- 21630155 TI - Cell cycle adaptations and maintenance of genomic integrity in embryonic stem cells and induced pluripotent stem cells. AB - Pluripotent stem cells have the capability to undergo unlimited self-renewal and differentiation into all somatic cell types. They have acquired specific adjustments in the cell cycle structure that allow them to rapidly proliferate, including cell cycle independent expression of cell cycle regulators and lax G(1) to S phase transition. However, due to the developmental role of embryonic stem cells (ES) it is essential to maintain genomic integrity and prevent acquisition of mutations that would be transmitted to multiple cell lineages. Several modifications in DNA damage response of ES cells accommodate dynamic cycling and preservation of genetic information. The absence of a G(1)/S cell cycle arrest promotes apoptotic response of damaged cells before DNA changes can be fixed in the form of mutation during the S phase, while G(2)/M cell cycle arrest allows repair of damaged DNA following replication. Furthermore, ES cells express higher level of DNA repair proteins, and exhibit enhanced repair of multiple types of DNA damage. Similarly to ES cells, induced pluripotent stem (iPS) cells are poised to proliferate and exhibit lack of G(1)/S cell cycle arrest, extreme sensitivity to DNA damage, and high level of expression of DNA repair genes. The fundamental mechanisms by which the cell cycle regulates genomic integrity in ES cells and iPS cells are similar, though not identical. PMID- 21630156 TI - Cell cycle regulation by microRNAs in stem cells. AB - The ability to self-renew and to differentiate into at least one-cell lineage defines a stem cell. Self-renewal is a process by which stem cells proliferate without differentiation. Proliferation is achieved through a series of highly regulated events of the cell cycle. MicroRNAs (miRNAs) are a class of short noncoding RNAs whose importance in these events is becoming increasingly appreciated. In this chapter, we discuss the role of miRNAs in regulating the cell cycle in various stem cells with a focus on embryonic stem cells. We also present the evidence indicating that cell cycle-regulating miRNAs are incorporated into a large regulatory network to control the self-renewal of stem cells by inducing or inhibiting differentiation. In addition, we discuss the function of cell cycle-regulating miRNAs in cancer. PMID- 21630154 TI - Function of the A-type cyclins during gametogenesis and early embryogenesis. AB - The cyclins and their cyclin-dependent kinase partners, the Cdks, are the basic components of the machinery that regulates the passage of cells through the cell cycle. Among the cyclins, those known as the A-type cyclins are unique in that in somatic cells, they appear to function at two stages of the cell cycle, at the G1 S transition and again as the cells prepare to enter M-phase. Higher vertebrate organisms have two A-type cyclins, cyclin A1 and cyclin A2, both of which are expressed in the germ line and/or early embryo, following highly specialized patterns that suggest functions in both mitosis and meiosis. Insight into their in vivo functions has been obtained from gene targeting experiments in the mouse model. Loss of cyclin A1 results in disruption of spermatogenesis and male sterility due to cell arrest in the late diplotene stage of the meiotic cell cycle. In contrast, cyclin A2-deficiency is marked by early embryonic lethality; thus, understanding the function of cyclin A2 in the adult germ line awaits conditional mutagenesis or other approaches to knock down its expression. PMID- 21630157 TI - Cell cycle regulation during proliferation and differentiation of mammalian muscle precursor cells. AB - Proliferation and differentiation of muscle precursor cells are intensively studied not only in the developing mouse embryo but also using models of skeletal muscle regeneration or analyzing in vitro cultured cells. These analyses allowed to show the universality of the cell cycle regulation and also uncovered tissue specific interplay between major cell cycle regulators and factors crucial for the myogenic differentiation. Examination of the events accompanying proliferation and differentiation leading to the formation of functional skeletal muscle fibers allows understanding the molecular basis not only of myogenesis but also of skeletal muscle regeneration. This chapter presents the basis of the cell cycle regulation in proliferating and differentiating muscle precursor cells during development and after muscle injury. It focuses at major cell cycle regulators, myogenic factors, and extracellular environment impacting on the skeletal muscle. PMID- 21630158 TI - Drosophila neural stem cells: cell cycle control of self-renewal, differentiation, and termination in brain development. AB - The wealth of neurons that make up the brain are generated through the proliferative activity of neural stem cells during development. This neurogenesis activity involves complex cell cycle control of proliferative self-renewal, differentiation, and termination processes in these cells. Considerable progress has been made in understanding these processes in the neural stem cell-like neuroblasts which generate the brain in the genetic model system Drosophila. Neuroblasts in the developing fly brain generate neurons through repeated series of asymmetrical cell divisions, which balance self-renewal of the neuroblast with generation of differentiated progeny through the segregation of cell fate determinants such as Numb, Prospero, and Brat to the neural progeny. A number of classical cell cycle regulators such as cdc2/CDK1, Polo, Aurora A, and cyclin E are implicated in the control of asymmetric divisions in neuroblasts linking the cell cycle to the asymmetrical division machinery. The cellular and molecular identity of the postmitotic neurons produced by proliferating neuroblasts is influenced by the timing of their exit from the cell cycle through the action of a temporal expression series of transcription factors, which include Hunchback, Kruppel, Pdm, and Castor. This temporal series is also implicated in the control of termination of neuroblast proliferation which is effected by two different cell cycle exit strategies, terminal differentiative division or programmed cell death of the neuroblast. Defects in the asymmetric division machinery which interfere with the termination of proliferation can result in uncontrolled tumorigenic overgrowth. These findings in Drosophila brain development are likely to have general relevance in neural stem cell biology and may apply to cell cycle control in mammalian brain development as well. PMID- 21630159 TI - Control of neuronal ploidy during vertebrate development. AB - Somatic tetraploid neurons are present in different structures of the vertebrate nervous system, including cortex and retina. In this chapter, we provide evidence that these neurons can be widely detected in the chick nervous system. We also discuss mechanisms creating neuronal tetraploidy in vertebrates, concluding that the neurotrophin receptor p75 could be responsible for the generation of these neurons in most neural tissues, as previously observed in the retina. Somatic tetraploidy in the chick retina correlates with increased neurons' soma size and dendritic arborization, giving rise to neurons known to innervate a specific layer of the optic tectum. Tetraploidy could therefore account for neuronal diversity in the normal nervous system. De novo generation of tetraploid neurons has been shown to occur in Alzheimer's disease. This suggests that the morphological changes expected to occur in the affected neurons could lead to altered neuronal function, thus providing a basis for neurodegeneration. PMID- 21630160 TI - Cell cycle deregulation in the neurons of Alzheimer's disease. AB - The cell cycle consists of four main phases: G(1), S, G(2), and M. Most cells undergo these cycles up to 40-60 times in their life. However, neurons remain in a nondividing, nonreplicating phase, G(0). Neurons initiate but do not complete cell division, eventually entering apoptosis. Research has suggested that like cancer, Alzheimer's disease (AD) involves dysfunction in neuronal cell cycle reentry, leading to the development of the two-hit hypothesis of AD. The first hit is abnormal cell cycle reentry, which typically results in neuronal apoptosis and prevention of AD. However, with the second hit of chronic oxidative damage preventing apoptosis, neurons gain "immortality" analogous to tumor cells. Once both of these hits are activated, AD can develop and produce senile plaques and neurofibrillary tangles throughout brain tissue. In this review, we propose a mechanism for neuronal cell cycle reentry and the development of AD. PMID- 21630161 TI - Sequential or concomitant chemotherapy in limited stage small-cell lung cancer. AB - PURPOSE: Chemotherapy (CT) combined with radiation therapy (RT) is the standard treatment for limited disease small-cell lung cancer (LDSCLC). Many questions including RT dose, fractionation, and sequence of RT/CT administration remain controversial. In this paper, we retrospectively assessed the outcome of patients with LDSCLC treated with radiation of at least 50 Gy. METHODS AND MATERIALS: From December 1997 to January 2006, 69 consecutive patients with LDSCLC were treated at our institutions. Treatment consisted of at least 4 cycles of CT, and 3D conformal thoracic RT. The median age was 61 years (range, 37-78 years). Sequential or concomitant CT/RT was given in 47 (68%) and 22 (32%) of the patients, respectively. The median RT dose was 60 Gy. Prophylactic cranial irradiation (PCI) was administered in 47 (68%) patients. RESULTS: With a median follow-up of 36 months (range, 6-107), 16 patients were alive without disease. The median overall survival time was 24 months, with a 3-year survival rate of 29%. The 3-year disease-free survival (DFS) and loco-regional control (LRC) rates were 23% and 60%, respectively. A better DFS was significantly associated with performance status (PS) 0 (p = 0.004), complete response to treatment (p = 0.03), and PCI group (p = 0.03). A trend towards improved overall survival (OS) was observed for patients who underwent PCI (p = 0.07). Patients treated with sequential CT/RT had a better outcome than those treated with concomitant treatment (3-year DFS rate 27% vs. 13%; p = 0.04). However, PCI was delivered more frequently for the sequential group. No significant dose-response relationship was found in terms of LRC. The multivariate analysis showed that complete response to treatment was the only significant factor for OS. CONCLUSION: Complete response to treatment was the most important factor for OS. A better DFS was significantly associated with the PCI group. We did not find a significant difference in outcome between patients receiving doses of 60 Gy or more and patients receiving 60 Gy or less. PMID- 21630162 TI - Implementation of an INR self-testing and self-management programme in common ambulatory private practice: our experience with 90 patients. AB - QUESTIONS UNDER STUDY: INR self-testing devices allow patients on vitamin K antagonists (VKA) to determine their INR and then have their VKA dose adapted by a physician (INR self-testing, ST) or adapt it themselves according to pre established guidelines (INR self-management, SM). The safety, efficacy and advantages of ST and SM have been demonstrated, but their use remains limited. In an effort to improve the availability of ST and SM, we tested the hypothesis that implementing a teaching programme for ST and SM in a small structure in common ambulatory private practice is feasible, safe and can lead to high patient satisfaction. METHODS: Patients on long-term anticoagulation were assigned to a specific training programme. Patients used the CoaguChek (S then XS) INR testing system. Technical problems, adverse events and INR values were then recorded during the first year of follow-up and analysed. Patient satisfaction data were obtained via a specific questionnaire. RESULTS: 169 patients were referred and 90 included in the teaching programme. 80 performed SM and 10 performed ST. 54 patients (60%) returned the 1-year questionnaire with complete INR data available for 35 patients. The percentage of INR in the target range (target +/- 0.5) was 60.6%. The rate of major adverse clinical events (MACE) was 3.7 per 100 patient years. The main reported advantages were a reduction in visits to the INR testing facility and increased autonomy. There was better venous to capillary INR correlation with the CoaguChek XS than with the S (p <0.025). CONCLUSIONS: The development in a small structure in common ambulatory practice of a specific teaching programme made ST and SM available to a new patient population. It led to high patient satisfaction, significantly reducing the burden of VKA monitoring. These results were obtained while preserving the safety and efficacy standards of VKA treatment and favour greater expansion of ST and SM programmes. PMID- 21630163 TI - Validation of prognostic factors and survival of patients with multiple myeloma in a real-life autologous stem cell transplantation setting: a Swiss single centre experience. AB - PRINCIPLES: High-dose chemotherapy with subsequent autologous stem cell transplantation (ASCT) is an important treatment option in younger patients with multiple myeloma (MM). We analysed the outcome of patients treated at our institution outside the clinical trials framework and tried to identify risk factors prognostic for survival. METHODS: Medical histories of the patients were screened for response, event-free survival (EFS) and overall survival (OS). Pre transplant variables were analysed to identify possible prognostic risk factors. RESULTS: Overall, 182 ASCT were performed in 120 patients with MM from 2002 to 2007. Treatment-related mortality (TRM) was 0.5%. Median EFS was 23.1 months (95% confidence interval [CI]: 19.4-28.4) and median OS was 49.8 months (95%CI: 43.7 - not reached) in the whole patient population. The median OS in patients who received one ASCT was 46.4 months (95%CI: 35.2 - not reached), and 63.7 months (95%CI: 48.9 - not reached) in patients who underwent double ASCT. Patients who already achieved a complete remission (CR) before ASCT had a longer EFS (p = 0.016) than patients without CR. Additionally, patients who achieved a CR after ASCT had a longer EFS (p = 0.0061) and OS (p = 0.0024) than patients without CR. ISS stage 34 + 0 were included in this case control study. The study period was 18 months. The last Doppler measurement in pregnancies with GDM (diet-controlled and insulin-dependent) was compared to the healthy control group. Our collected data included the last prenatal Doppler flow recordings (resistance index (RI) in the umbilical artery (UA)). RESULTS: In women with diet-controlled GDM, a significant decrease in the RI (p = 0.002) in the UA has been observed. Insulin-treated diabetic and healthy control pregnancies showed no difference in the RI. CONCLUSION: Doppler flow examinations with RI measurements in patients with GDM differ significantly with respect to healthy controls. In insulin-treated women the RI indices are not different from the control group, while in the diet-controlled group a significant decrease was noted and additionally might show a possible maternal metabolic dysfunction. PMID- 21630181 TI - [Automated breast ultrasound vs. handheld ultrasound: BI-RADS classification, duration of the examination and patient comfort]. AB - PURPOSE: Automated breast ultrasound (ABUS) is a potentially valuable adjunct to mammography in breast cancer screening. The reliability and the inter-observer variability in the BI-RADS classification, compared to handheld ultrasound (US), as well as the duration of the examination and patient comfort have only been investigated in a limited number of papers to date. MATERIALS AND METHODS: In a prospective study, we examined 148 breasts of 76 patients with handheld US and ABUS. The ABUS data were evaluated separately by two investigators. Patient comfort was assessed using a standardized questionnaire. RESULTS: The inter observer agreement for the BI-RADS classification among the two observers using ABUS was high (kappa = 0,750), the agreement with handheld US was moderate. The sensitivity in the detection of breast cancer was 87.5 % for handheld US and 75 % for the ABUS evaluation by observer 1. The sensitivity was 87.5 % for the ABUS evaluation and 83 % for mammography by observer 2. The ABUS examination was rated as completely painless by 64 % of the patients. 25 % of the patients indicated minor pain, and 10 % indicated moderate pain. Handheld US was rated as completely painless by 66 % of the patients. 26 % of the patients indicated minor pain, and 8 % indicated moderate pain. CONCLUSION: ABUS examinations focusing on the BIRADS classification have low inter-observer variability, compared to handheld US. The sensitivity of ABUS did not differ significantly from handheld US. PMID- 21630182 TI - Assessment of fetal global and regional ventricular function in congenital heart disease using a novel feature tracking technique. AB - PURPOSE: The aim of this study was to assess global and regional longitudinal peak systolic ventricular function in fetuses with congenital heart disease (CHD) and compare measurements derived from feature tracking with reference values of healthy fetuses with a matching gestational age. MATERIALS AND METHODS: Global and segmental longitudinal peak systolic strain (LPSS), strain rate and velocity values of the left (LV) ventricular myocardium and right (RV) ventricular myocardium were assessed by a novel feature tracking technique in 17 fetuses with congenital heart disease (10 fetuses with left heart obstruction (LHO), 5 fetuses with tetralogy of Fallot (TOF) and 2 fetuses with double outlet right ventricle (DORV)) and were compared in a second step with 24 matched healthy fetuses (gestational age range 21 - 36 weeks of gestation). RESULTS: The global LPSS of both ventricles was slightly elevated in fetuses with TOF or DORV compared with controls (RV: p = 0.055; LV: p = 0.063). The RV strain rate presented a trend toward higher values (p = 0.09). Corresponding global LV velocity values of fetuses with TOF or DORV revealed a tendency to decrease compared to healthy controls (p = 0.054). In contrast, all measurements of fetuses with LHO did not show any statistical difference regarding deformation parameters or velocity compared to healthy controls. CONCLUSION: Global longitudinal peak systolic measurements in fetuses with congenital heart disease revealed slightly higher strain (RV/LV) and strain rate (RV) values in the subgroup of fetuses with TOF or DORV compared to healthy controls. Whether the application of this new angle independent technique is suitable for discriminating between healthy and diseased fetuses has to be verified in further investigations. PMID- 21630183 TI - Intraobserver and interobserver reliability of transvaginal cervical length measurements and quantitative ultrasound tissue characterization of the cervix in the second and third trimester of pregnancy. AB - PURPOSE: To determine the intraobserver and interobserver reliability of transvaginal ultrasound measurements of cervical length and quantitative ultrasonic tissue characterization (QUTC). MATERIALS AND METHODS: 20 healthy women in the second and third trimester of pregnancy underwent transvaginal ultrasound examination of the cervix by two sonographers. 3 replicate vaginal measurements of cervical length and QUTC were taken by each observer for each woman. The repeatability coefficient and intraclass correlation coefficient as well as the interclass correlation coefficient and limits of agreement were calculated. RESULTS: The reliability coefficient for transvaginal cervical length measurements was 0.95 for sonographer 1 and 0.99 for sonographer 2. The corresponding intraclass correlation coefficients for cervical length measurements were 0.872 and 0.973. The interclass correlation coefficient for transvaginal cervical length measurements was 0.986, the mean interobserver difference was -2 mm, and the limits of agreement were -5.4 mm to 1.5 mm. The reliability coefficients for QUTC were 0.90 (sonographer 1) and 0.97 (sonographer 2) and the corresponding intraclass correlation coefficients for cervical length measurements were 0.74 and 0.9. The interclass correlation coefficient was 0.56, the mean interobserver difference was 0.14, and the limits of agreement were 22.46 to 22.74. CONCLUSION: Transvaginal cervical length measurements showed a high intraobserver and interobserver reliability, while quantitative ultrasonic tissue characterization had poor interobserver reliability and should currently not be applied in clinical practice. PMID- 21630184 TI - Diagnostic value of quantitative EUS elastography for malignant pancreatic tumors: relationship with pancreatic fibrosis. AB - PURPOSE: EUS elastography has been used to facilitate the diagnosis of pancreatic cancer, but as yet the interpretation of this procedure has been largely subjective. The present study has been designed to validate a quantitative approach for the analysis of EUS elastography, and to assess its relationship with pancreatic fibrosis. MATERIALS AND METHODS: 86 patients with malignant pancreatic masses and 28 control subjects without any evidence of pancreatic diseases were examined by EUS elastography. EUS video sequences were subjected to a quantitative analysis based on mean hue histogram analysis. Pancreatic fibrosis was determined by quantitative morphometry in tissue specimens from 36 patients. RESULTS: The mean RGB (red, green, blue) value was significantly higher in the cancer patients compared to the controls (14.0 +/- 0.4 vs. 11.5 +/- 0.9; p = 0.0085), albeit with significant overlap between the groups. In contrast, a much sharper separation between the groups was obtained based on the individual color values for blue, green and red (p < 0.0001, respectively). By these means, 100 % sensitivity and specificity for the distinction between tumor and normal tissue was obtained for the blue color value, while the red and green color values were less discriminative. The fractional fiber content of the tumors was unrelated to the respective hue histogram color values. CONCLUSION: Quantitative EUS elastography allows for clear differentiation between malignant pancreatic tumors and normal tissue. Using this approach, we demonstrated that the stiffness of pancreatic tumors is largely independent of their fiber content. PMID- 21630185 TI - [Contrast-enhancend sonography (CEUS) in pneumonia: typical patterns and clinical value - a retrospective study on n = 50 patients]. AB - PURPOSE: To describe contrast-enhanced ultrasound (CEUS) patterns of pneumonia, to characterize CEUS patterns and to determine the clinical value of deviant CEUS patterns. PATIENTS AND METHODS: N = 50 patients with radiologically diagnosed alveolar pneumonia were investigated by CEUS and retrospectively evaluated. Pulmonary enhancement was differentiated from bronchial arterial enhancement by measurement of time to enhancement from the application of the contrast agent (CA). The echogenicity of the CA enhancement was evaluated (isoechoic/hypoechoic) using the spleen as an "in vivo reference". In addition, the homogeneity of the CA enhancement (homogeneous/ inhomogeneous) was recorded. The patients were divided into two groups according to the CEUS pattern (type 1/type 2) and compared to each other in terms of age, days of hospitalization, comorbidity, rate of complications and the presence of pleural effusion. RESULTS: The majority showed a type 1 CEUS pattern consisting of a pulmonary arterial supply (92 %), an isoechoic extent of enhancement (74 %) and a homogeneous enhancement (78 %) of the CA in the pulmonary lesions. The only significant difference found between the two groups was the average age. CONCLUSION: Pneumonia most likely shows a type 1 CEUS pattern consisting of a pulmonary arterial supply, an isoechoic extent of enhancement compared to the spleen and a homogeneous enhancement of the CA in the pulmonary lesions. Prognostic value of a type 2 CEUS pattern (bronchial arterial supply and/or hypoechoic extent of enhancement and/or inhomogeneous enhancement) in pneumonia regarding days of hospitalization, comorbidity, rate of complications and the presence of pleural effusion could not be shown. PMID- 21630186 TI - [Diagnostic value of transthoracic ultrasound compared to chest radiography in the detection of a post-interventional pneumothorax]. AB - PURPOSE: Chest radiography is standard for the diagnosis of a pneumothorax. However, also ultrasound of the chest has considerable value in the detection of a pneumothorax. A typical sonographic feature is the lack of the lung gliding sign. Aim of our trial was to evaluate the significance of ultrasound of the chest in the diagnosis of a pneumothorax supplemented by using power colour Doppler imaging. PATIENTS AND METHODS: Following transbronchial biopsy, patients received transthoracic ultrasound of the chest for evaluation of a pneumothorax. Immediately afterwards, a chest radiograph was performed and results were compared. RESULTS: 1023 patients (321 female, 702 male, median age 47 years) were examined. In 30 patients (2.9%) chest radiograph revealed a pneumothorax, while in ultrasound of the chest a pneumothorax was diagnosed in 36 of the cases. Defining chest radiography as gold standard, ultrasound of the chest had a sensitivity of 100%, a specificity of 83% and an accuracy of 99%. CONCLUSION: Transthoracic ultrasound of the chest is a highly sensitive and specific diagnostic tool in the diagnosis of a pneumothorax. In comparison to chest radiography, it is better available and prevents administration of ionizing radiation. However, a disadvantage of ultrasound is the lack of quantification of a pneumothorax and the assessment of the indication for chest tube drainage. PMID- 21630187 TI - New sonographic method for fetuses with small abdominal circumference improves fetal weight estimation. AB - PURPOSE: Accurate estimation of fetal weight is a valuable tool for determining further obstetric management. Commonly used weight formulas lack accuracy, even though some equations appear to be favorable within defined weight ranges. However, due to the fact that fetal weight is not known in advance, it is not always clear which formula is suitable. In most of the commonly used equations, the fetal abdominal circumference (AC) is not only included but also has the greatest impact on weight estimation. The aim of our study was to develop and evaluate a new formula specifically designed for a small fetal AC in order to improve weight estimation. MATERIALS AND METHODS: The study included 323 pregnancies. The inclusion criteria were singleton pregnancy, ultrasound examination with complete biometric parameters and an AC <= 29.0 cm within 7 days of delivery, and an absence of structural or chromosomal malformations. Two "best fit" formulas were derived by forward regression analysis. Finally, the accuracy of the new formulas was compared to commonly used weight equations by using the percentage error, absolute percentage error (APE), limits of agreement (LOA) and cumulative distribution. RESULTS: Contrary to the routine methods, which significantly underestimated fetal weight, the new formulas did not have a systematic error. The medians of the APE were the lowest (7.13 and 7.16) when compared to other equations. Moreover, the new formulas demonstrated the narrowest LOA. At all discrepancy levels (5%, 10%, 15%, and 20%), the new formulas included significantly more cases than the commonly used methods. CONCLUSION: The specifically designed equations help to improve fetal weight estimation for fetuses with an AC <= 29.0 cm. For optimal weight estimation, we recommend using the new formula II. PMID- 21630188 TI - Course and outcome of pregnancy after maternal exposure to angiotensin-II receptor blockers--case report and review of the literature. PMID- 21630189 TI - Treatment results in patients with intracranial ependymomas. AB - OBJECTIVE: To evaluate the treatment outcomes of patients with intracranial ependymomas. METHODS: Between 1988-2007, 27 consecutive patients, with 9 patients (33%) aged under 16 years, were treated at our institution for an intracranial ependymoma. Pertinent clinical data were retrieved from the patients' charts. The histopathological findings in 25 cases were reviewed using the 2007 World Health Organization (WHO) classification system. Median follow-up was 84 months. RESULTS: Infratentorial tumors were diagnosed in 22, and supratentorial tumors in 5 patients. Histopathological findings were ependymoma WHO grade II (E II) in 14 patients, and anaplastic ependymoma WHO grade III (AE III) in 13 patients. A complete tumor resection was achieved with the first operation in 20 cases (74%). Primary adjuvant therapy consisted of chemotherapy alone in a 17-month-old child with an incompletely resected posterior fossa AE III, radiation therapy alone in 4 cases, and combined radiation therapy and chemotherapy in 7 cases. Tumor recurrence was seen in 10 cases (37%), including 5 patients with an E II and 5 patients with an AE III. The 5-year progression-free survival (PFS) was 74% and 67% for E II and AE III, respectively. The 5-year PFS was 80% following a complete resection, and 56% in patients with a residual tumor. CONCLUSIONS: Surgery alone, as the primary treatment, achieves a good outcome in most patients with E II. Good results can be achieved with surgery and adjuvant local radiotherapy in patients with AE III. PMID- 21630190 TI - Sustained relief after discontinuation of DBS for dystonia: implications for the possible role of synaptic plasticity and cortical reorganization. PMID- 21630191 TI - [Violation Type III acromioclavicular joint - a challenge even for the physical therapy!]. PMID- 21630193 TI - [Stem cell--is there any role in tumorigenic activity]. AB - Stem cells are a quintessential key to proper behavior of homeostatic processes. They are often thought of as the solution to a wide range of human conditions, with the ability to rescue malfunctioning or non-functioning organs and tissues. However, there is increasing evidence that stem cells can play a central role in disease. Most recently stem cells have been implicated in cancer after not responding to homeostasic controls such as proliferation and differentiation. Cancer has long been seen as a disease that arises from mutations that impair the capacity of any cell within the organism to respond to the signals that regulate proliferation. Besides their scarcity or abundance, a second important issue with respect to cancer stem cells is their origin. A new defining model for carcinogenesis, the "cancer stem cell hypothesis" was put forward in 2006, according to which cancer is a stem cell disease that places malignant stem cells at the centre of its tumorigenic activity as they have the capacity to undergo self-renewal, and have the potential to differentiate into different types of cells in a specific lineage. PMID- 21630194 TI - [Pathology laboratories staff workload evaluation in Turkey: a survey study]. AB - OBJECTIVE: The workload affects the quality of the pathology report. The aim of this study was to investigate the territorial distribution and productivity of pathology laboratories around Turkey and to estimate the staff workload. MATERIAL AND METHOD: A survey questioning the workload was sent to all Ministry of Health and university hospitals. Staff workload was questioned according to the hospital classification and educational activity to evaluate the productivity. Data were entered using SPSS 16.0 statistical software package program and the distribution criteria, t-test and one-way anova were used in the analysis to evaluate the differences between the averages. RESULTS: An average of 2.8 pathologists worked at the pathology laboratories. A total of 5.500 biopsies and 3.750 cytology specimens were received and 20.000 blocks prepared per year. Pathologists evaluated 1.935 biopsies and 1.400 cytology specimens on average and this is equivalent to 2.718 biopsies per year. Gynecology and general surgery department materials constituted 57 percent of all biopsies. Each technician prepared 6.200 blocks, 11.500 slides and 1.000 immunohistochemistry preparations on average. An average of 3.4 paraffin blocks was prepared for each biopsy. The efficiency was low in 17% of teaching hospitals and 77.8% of non-teaching hospitals. In contrast 62.5% of teaching hospitals had work overload. The majority (70.5%) of the respondents mentioned staff shortage. CONCLUSION: There is no pathologist shortage in Turkey and the problem is workload distribution. Pathology residents' overwork would be reduced by using pathology assistants. There is no shortage of technicians or secretaries, but uneven distribution. Pathology staff planning must be tailored taking into account the features of each hospital. Standard planning for all hospitals is not suitable. PMID- 21630195 TI - [A new remedy in pathology practice: molecular solution to sample mix-up]. AB - OBJECTIVE: Although very rare, suspicious situations about the identity of diagnostic tissue material have been encountering in pathology practice. Such situations undoubtedly have the potential to create undesirable results. In the present study, an application targeting getting rid of any doubts about the identity of the diagnostic tissue samples is described. MATERIAL AND METHOD: A combination of short tandem repeats (STR) of the human genome consisting of CSF1PO, TH01, TPOX, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539 and Penta E were selected on the basis of ease of application and bioinformatic discrimination power. Possible forms of diagnostic tissue mix up were set in 3 different models with 3 diagnostic tissue samples of 2 different cases. Of the tissue samples selected, A (salivary gland) and B (striated muscle) belonged to the same case and C (uterus wall) belonged to another case. In the first model, there was no problem about tissue identity (M1: A/B). In the second model, two different diagnostic material were mixed up (M2: B/C). In the last model, there were 3 diagnostic material obtained from 2 different cases (M3: A/B/C). DNA was extracted from all tissue samples and all of the selected 10 STR were amplified with specially designed primers by PCR. After chemical denaturation, amplicons were submitted to polyacrylamide gel electrophoresis for discrimination of single DNA strands according to their conformation polymorphism (SSCP). Special patterns of each STR in the gel matrix obtained from M1, M2 and M3 models, were evaluated on the principle of being 'same or different' to determine the diagnostic material identity. RESULTS: Each of the salivary gland, striated muscle and uterus wall samples were correctly identified (matched with the right source cases) after evaluating 10 different STR SSCP patterns designed under M1, M2 and M3 models. CONCLUSION: This application targeting to solve diagnostic tissue identity problems is a simple and cheap application of SSCP and its efficacy was proven on the designed models. PMID- 21630196 TI - CD138 expression in renal tumors and its usage in the differential diagnosis. AB - OBJECTIVE: The differential diagnosis of kidney tumors, especially those with eosinophilic cytoplasms, can be problematic due to overlapping morphologic features. CD138 is primarily a plasma cell marker but is known to be expressed in the proximal renal tubular epithelium as well. This study aims to investigate the possible contribution of CD138 expression in the differential diagnosis of kidney tumors with eosinophilic cytoplasm. MATERIAL AND METHOD: The case series consisted of 15 chromophobe (ChRCC), 5 eosinophilic variant (EoRCC), 10 clear cell (CCRCC) and 9 papillary (PRCC) renal cell carcinomas, and 13 oncocytomas. Sections obtained from representative paraffin blocks were stained against CD138 antibody. RESULTS: All CCRCC and PRCC showed membranous CD138 expression. In some of the other eosinophilic renal tumors, cytoplasmic CD138 labeling in varying degrees was detected. In CCRCC cases, CD138 expression was especially observed in low grade areas and areas showing cystic and pseudopapillary growth patterns. A similar pattern of cytoplasmic staining was seen in 3 of the EoRCC and the most of the PRCC cases (6/9). CONCLUSION: Our findings suggest that CD138 may contribute to the differential diagnosis of renal tumors because of the membranous staining pattern in CCRCC and EoRCC cases and the cytoplasmic staining in CHRCC and oncocytoma cases. Its contributory role may be improved by combined usage with markers like Cytokeratin 7 and RCC marker. PMID- 21630197 TI - Mucinous tubular and spindle cell carcinoma of kidney and problems in diagnosis. AB - OBJECTIVE: Mucinous tubular and spindle cell carcinomas (MTSCC's) are recently described rare type of renal cell carcinoma (RCC). MTSCC's are characterized by small, elongated tubules lined by cuboidal cells and/or cords of spindled cells separated by pale mucinous stroma. They have morphological similarities to papillary RCC (papRCC). We evaluated the importance of the immunohistochemical features in the differential diagnosis of MTSCC and papRCC. MATERIAL AND METHOD: We re-evaluated 9 cases of MTSCC diagnosed between 2004 and 2010 and compared 10 cases of papRCC. All tumors were stained with alpha-methylacyl-CoA racemase (AMACR), cytokeratin 7 (CK7), CK19, renal cell carcinoma marker (RCC Ma), CD10 and kidney specific cadherin (KspCad). RESULTS: A total of 6/9 cases were considered classical. Two of 9 MTSCC's were classified as "mucin-poor". Foamy macrophages were identified in 4 cases. The immunoreactivity in MTSCC was AMACR 100%, CK7 100%, CK19 100%, RCC Ma 50%, CD10 11%, and KspCad 38% while the values for papRCC were AMACR 100%, CK7 90%, CK19 100%, RCC Ma 100%, CD10 80%, and KspCad 0%. CONCLUSION: MTSCCs may include little mucin and show a marked predominance of either of its principal morphological components. They may mimic other forms of RCC. Pathologists should be aware of the histological spectrum of MTSCCs to ensure an accurate diagnosis. Careful attention to the presence of a spindle cell population may be helpful in the differential diagnosis in tumors with predominant compact tubular growth. Immunohistochemical stains for papRCC are also expressed in MTSCC, but strong CD10 expression may not favor MTSCC. PMID- 21630198 TI - Expression of CD44 and MMP-2: possible association with histopathological features of pleuro-pulmonary solitary fibrous tumors. AB - OBJECTIVE: Recent studies have shown that tumor cell adhesion molecules CD44 and matrix metalloproteinases (MMP-2) are expressed strongly in many tumors and associated closely with invasion and metastasis of these tumors. Although solitary fibrous tumors (SFT) have a good prognosis, a minority behave malignantly. The aim of this study was to analyze the correlation between CD44 and MMP-2 expression with histopathological parameters in SFT. MATERIAL AND METHOD: Haemotaxylin-Eosin stained sections of 10 patients with SFT were reexamined for evaluation of histopathological parameters. Immunostaining of CD44 and MMP-2 was performed by using the streptavidin-biotin method with mouse monoclonal antibody. RESULTS: Our cases consisted of three male and seven female patients with a mean age of 54.5 years. Three patients had a history of asbest exposure. Complete resection was performed in 2 malignant (multiple masses) and 8 benign SFT cases. One intrapulmonary tumor was treated with pneumonectomy. 3 cases originated from the right and 7 from the left hemithorax. Tumor size ranged from 5 to 27cm. All cases expressed strong CD44. Only 2 malignant SFT and intrapulmonary SFT expressed focal MMP-2. CONCLUSION: Although MMP-2 positivity was observed in 2 malignant cases, CD44 positivity was not associated with malignancy criteria in solitary fibrous tumors. PMID- 21630199 TI - Histopathological examination of patients operated on for a neck mass: 4-year follow-up results. AB - OBJECTIVE: To investigate the diagnostic distribution of neck masses treated surgically at our clinic and also to discuss the clinical presentation and histological finding of these pathologies. MATERIAL AND METHOD: We retrospectively reviewed the records (age, sex, clinical presentation and histological findings) of 201 patients who presented at our clinic with neck masses between 2006 and 2010. RESULTS: Of the 201 patients, 98 (48.75%) were classified as inflammatory masses, 67 (33.33%) as neoplastic neck masses, and 36 (17.91%) as congenital neck masses. The mean age was 27.2+/-17.2 in patients with inflammatory neck masses and 11.4+/-4.6 for congenital neck masses. In the neoplastic neck masses group, the mean age was 37.4+/-11.6 for benign lesions but 49.4+/-20.6 in malignant lesions. CONCLUSION: When a neck mass is seen, neoplasms should be considered in older adults and inflammatory and congenital masses in children and young patients. Although the history, medical examination and additional diagnostic methods provide important information, the exact diagnosis may only be obtained by histopathological examination. PMID- 21630200 TI - Low-grade osteosarcoma, review of 15 cases in a series of 156 osteosarcoma cases. AB - OBJECTIVE: Osteosarcoma of the bone accounts for approximately 20% of all primary malignant bone tumors. Most cases (75-85%) are high-grade osteosarcomas. We aimed to introduce low-grade osteosarcomas that we found by retrospectively scanning our archives because of the diagnostic difficulties, rarity and different therapeutic approach required. MATERIAL AND METHOD: We found 156 osteosarcoma cases in the archives of the Department of Pathology of Istanbul University, Cerrahpasa Faculty of Medicine covering the period 2000 to 2010. These cases included 141 high-grade and 15 low-grade osteosarcomas. RESULTS: Low-grade osteosarcomas made up 10.4% of all osteosarcomas. Age and sex distribution were 19 to 54 (median 33.6) with 9 females and 6 males. Most cases were located in the distal femur while others were in the proximal tibia, the left wrist, the distal tibia and the right knee. The tumors were generally made up of relatively well formed trabeculae of woven bone surrounded by a moderately cellular spindle cell proliferation entangled in collagen fibers. Thirteen cases were reported as parosteal osteosarcoma, one as intramedullary well-differentiated osteosarcoma, and one as low-grade osteosarcoma metastasis in the lung. CONCLUSION: Low-grade osteosarcomas account for approximately 3-5% of all osteosarcomas. They are more commonly seen in women and 9 cases out of 6 were female in our series. The average age was 33.6 (19-54). The distal femur was the most common localization as is typical in osteosarcomas. In contrast to high-grade lesions, surgery will be sufficient in low-grade osteosarcomas. The survival rate ranges from 90% to 100%. Differentiating low-grade osteosarcomas from high-grade osteosarcomas and benign lesions and using a multidisiplinary approach for the diagnosis, treatment and follow-up periods are of vital importance. PMID- 21630201 TI - Histopathological correlation of squamous cell abnormalities detected on cervical cytology. AB - OBJECTIVE: To investigate the correlation between cytology and cervical biopsy in patients with squamous cell abnormality on cervical cytology. MATERIAL AND METHOD: The cervical smears diagnosed in our clinic between 2005-2008 were reviewed retrospectively. Cases which exhibited squamous cell abnormality (n: 374) were evaluated. RESULTS: The mean age was 45.15+/-10.78. In the cytopathological results, 256 (68.4%) ASC-US, 21 (5.6%) ASC-H, 31 (8.2%) LSIL, 48 (12.8%) HSIL, and 8 (4.8%) invasive carcinomas were diagnosed. Histopathological results were 213 (57%) nonneoplastic, 85 (22.7%) CIN I, 14 (3.7%) CIN II, 34(9.0%) CIN III and 28 (7.5%) invasive squamous cell carcinomas. Including all squamous cell abnormalities, the sensitivity of the smear test in CIN I and higher grade lesions was 56.95% and the false positivity was 43.04%. Excluding ASC-US and ASC-H lesions, the sensitivity of the smear test was 77.31% and the false positivity was 22.68%. After evaluating cervical cytohistopathological correlation, the positive predictive value was found to be 100% in invasive carcinoma, 62% in HSIL and 38% in LSIL. CONCLUSION: As the grade of cytopathological result increases, the correlation between biopsy and the smear test also increases. The high sensitivity of the cervical smear test for high grade lesions shows that it is an effective screening test. PMID- 21630202 TI - Diagnostic value of transbronchial needle aspiration cytology in sarcoidosis. AB - OBJECTIVE: Sarcoidosis is a systemic granulomatous inflammation that occurs as a result of disturbed immune regulation in individuals exposed to certain environmental agents. Although tissue sampling is considered the "gold standard" for the diagnosis of sarcoidosis, a medically treated disease, minimally invasive diagnostic methods are preferred instead of surgical tissue sampling. Transbronchial needle aspiration has been reported as an effective diagnostic method. MATERIAL AND METHOD: In this study, transbronchial needle aspiration cytology samples were assessed from 38 patients where sarcoidosis was suspected clinically and radiologically. The existence of epithelioid histiocytes and/or giant cells that formed granulomas was used as a cytological diagnostic criterion for chronic granulomatous inflammation. The presence of lymphocytes and/or germinal center cells, and of histiocytes in lymph nodes was regarded as adequate sampling criteria. RESULTS: A total of 31 out of the 38 patients were diagnosed as sarcoidosis with clinical, radiological and microbiological findings, after chronic granulomatous inflammation was considered by cytologic assessment. Cytologic diagnosis was achieved in: 4 of 7 patients with sampling from a single lymph node region, 25 of 28 patients with sampling from two different lymph node regions and 2 of 3 patients with sampling from three different lymph node regions. Two of the 7 patients who could not be diagnosed cytologically underwent a transbronchial parenchyma biopsy and the rest were diagnosed histologically from mediastinoscopic lymph node sampling. CONCLUSION: We would like to emphasize that transbronchial needle aspiration is a successful diagnostic method. We highlighted the adequacy criteria of cytological sampling and the encountered cytological findings of chronic granulomatous inflammation. PMID- 21630203 TI - An unusual case of thyroid papillary carcinoma with solitary cerebral metastasis presenting with neurological symptoms. AB - Papillary carcinoma of the thyroid is well known for metastasis to regional lymph nodes resulting in early detection and overall favorable prognosis. Solitary metastasis of papillary carcinoma of the thyroid in the cerebrum is extremely rare with only a few cases reported in the literature. Here, we report an unusual case of a 40-year-old female patient who presented with neurological symptoms and a radiological diagnosis of meningioma was made. Subsequent histopathology from the resected brain tumor revealed a solitary cerebral metastasis from papillary carcinoma of the thyroid. There was no involvement of regional lymph nodes and the radiological findings were indicative of a primary brain tumor. PMID- 21630204 TI - Cytopathological dilemma of anaplastic sacral chordoma with radiological and histological corroboration. AB - Chordoma is a relatively rare locally invasive and potentially malignant tumor of fetal notochord origin, affecting the axial skeleton. Cytopathological diagnosis of chordoma is favored by the presence of characteristic physaliphorous cells, bearing abundant foamy cytoplasm dispersed in a myxoid matrix. Anaplastic chordoma or dedifferentiated chordoma, an even rarer variant, can cause a diagnostic confusion with chondrosarcoma from the cytopathological point of view, with similar chondromyxoid matrix and atypical cells. Hence, chordoma bearing anaplastic features needs to be identified and should be distinguished from chondrosarcoma on aspiration cytopathology. We present a case of anaplastic sacral chordoma in a man 59 years of age, causing extensive destruction of sacrum and invading the paravertebral tissues as evidenced by radiology. Fine needle aspiration cytopathology revealed few large pleomorphic hyperchromatic cells, admixed with characteristic physaliphorous cells and myxoid matrix. The cytopathological diagnosis has been confirmed by histopathology and immunohistochemistry. Since anaplastic chordoma bears an unfavorable prognosis, it should be suspected on preoperative aspiration cytopathology. Clinicoradiological correlation along with histopathological and immunohistochemical confirmation is necessary subsequently. PMID- 21630205 TI - Gingival granular cell tumor of the newborn: a case report and review of literature. AB - The etiology and histogenesis of granular cell tumor are still debated. Granular cell tumor of the newborn is considered to be a different entity than the adult form of this lesion with different immunohistochemical features. We present a case of a rare gingival granular cell tumor in a newborn and review the literature. Gingival granular cell tumor must be clinically differentiated from teratoma, congenital dermoid cyst, congenital fibrosarcoma, hemangioma, lymphangioma, leiomyoma, rhabdomyoma, heterotopic gastrointestinal cyst, congenital cystic choristoma and congenital lipoma. Surface ulceration or pseudoepitheliomatous hyperplasia may lead to confusion with malignancy. PMID- 21630206 TI - Sarcomatoid chromophobe renal cell carcinoma with osteosarcoma-like differentiation which presented as a retroperitoneal mass: a case report. AB - Sarcomatoid component can occur in all histological subtypes of renal cell carcinoma but most commonly accompanies chromophobe renal cell carcinoma. The majority of the sarcomatoid components consist of malignant fibrous histiocytoma, fibrosarcoma or undifferentiated sarcoma areas. Heterologous differentiation in the form of osteosarcoma or rhabdomyosarcoma is very rarely encountered. We report a very rare case of a chromophobe renal cell carcinoma with osteosarcomatous differentiation that presented as a retroperitoneal mass. Renal cell carcinoma with sarcomatoid change should be always kept in mind when a retroperitoneal mass with sarcomatoid differentiation is encountered. PMID- 21630207 TI - Primary malignant melanoma of the ovary: case report and review of the literature. AB - Ovarian malignant melanomas are extremely rare tumors. Most of them are secondary tumors and disseminated metastases are recognized at the time of diagnosis. Primary tumors are even more rare and usually associated with a teratoma. A 67 year-old female had a pelvic mass that was recognized on ultrasonography (USG) and physical examination. Intraoperative pathological consultation was reported as "pigmented solid ovarian tumor, probably compatible with malignant melanoma". Paraffin sections, and histochemical (Masson Fontana and Prussia blue) and immunohistochemical examination (S-100 and HMB-45) were also consistent with "malignant melanoma". This case was accepted as "Probably primary ovarian malignant melanoma" in lack of any other tumor focus on detailed clinical and radiological investigation, skin biopsies or pigmented lesions in medical history. It is reported for being an extremely rare tumor and its distinctive characteristics for differential diagnosis are emphasized. PMID- 21630208 TI - Parachordoma: a recurrent case and review of the literature. AB - Parachordoma is an uncommon tumor of soft tissue, and the origin is not clear. Recurrence and metastasis are rarely seen. A piecemeal mass measuring 7x4x3 cm was excised from a 28-year-man who had presented with pain and swelling of the right shoulder for 5 years. Histopathologically, the tumor was composed of cells with clear eosinophilic cytoplasm and an epithelioid appearance in a myxoid stroma separated by fibrous tissue with mild pleomorphism and mitotic activity. Tumoral cells were immunoreactive for cytokeratin 8/18, EMA, S-100 and vimentin, immunohistochemically. Recurrence was seen one year after the initial diagnosis. Areas of increased mitotic activity and atypical mitoses were observed in the recurrent tumor. We report this case as recurrence occurred earlier than usual and exhibited malignant features. PMID- 21630209 TI - Histopathology in iraq: reliable diagnostics in spite of shortages. AB - The purpose of this study is to understand the status of Pathology in Iraq evaluating health problems in the country. Pathologists in Iraq are at the top of the level of scientific knowledge and professional responsibility. Although there are adequate modern equipment, lack of chemicals and experienced technical service inadequacies are evident. Special stains and immunohistochemistry are performed only in some of the private laboratories. Consultation is often carried out within the department, but external consultations are less frequent. In terms of specialized training, highly motivated, knowledgeable rotators and residents are available. Technical personnel, even though not sufficient in number, have got adequate professional training and skills. There are many problems in running quality control programs in the laboratories. As a result, the progress in the health system of Iraq, and therefore the status of pathology discipline, are of great importance for the development of the country. Foreign pathologists as a volunteer or a staff will provide the objective points of identifying the problems and will help to find a solution. PMID- 21630210 TI - Evaluation of CT as a predictor for kidney and renal artery mobility. AB - PURPOSE: Renal artery stent failure may result from excessive kidney mobility in some patients. We used computed tomography (CT) to determine the prevalence and magnitude of renal displacement due to postural changes. MATERIALS AND METHODS: A retrospective review of 100 consecutive CT colonography examinations was performed to measure renal artery location and displacement in both axial and coronal views using paired supine and prone non-contrast scans. Kidney displacement from the prone to supine position was correlated with a change in renal artery angular deviation. Statistical significance was determined using t tests and Pearson correlations. Results were based on measurements made by a single observer. RESULTS: Mobility and angular displacement between the prone and supine positions were significant bilaterally and in both planes (P < 0.01) except for the coronal plane kidney mobility on the left (P = 0.32). The axial plane correlation between kidney and artery mobility was significant bilaterally (left/right R=0.44/0.22, P < 0.01/0.03); the coronal plane correlation was only significant on the left (left/right R=0.26/0.18, P = 0.01/0.08). The mean axial plane mobility and angle change were greater on the left (left/right mobility 13 mm/7 mm; left/right angle change 18 degrees /8 degrees ). In contrast, the mean coronal plane mobility and angle change were greater on the right (left/right mobility 4 mm/22 mm; left/right angle change 4 degrees /8 degrees ). Fourteen patients had a mobility in excess of 32 degrees . CONCLUSION: During postural changes, the kidneys and renal arteries demonstrate significant correlated mobility. Renal artery movements can be identified using a low-radiation dose CT exam. PMID- 21630211 TI - Immunohistochemical study of the apoptosis process in epidermal epithelial cells of rats under a physiological condition. AB - Epidermal homeostasis is maintained by both epithelial proliferation in the stratum basale (SB) and the apoptosis of epithelial cells under physiological conditions. In this study, the induction and regulation mechanisms of epidermal apoptosis were immunohistochemically investigated in the epidermis from Wistar rat's palm and foot pad by using several apoptotic related proteins under a physiological condition. The results showed that Fas and Fas-L were expressed in cellular membranes of the stratum spinosum (SS), whereas TNF-R1 did not show any membranous expression in any epidermal layers. TNF-alpha was not observed in the epidermis. Caspase-10, cleaved caspase-3 and DNase-1 were found in the epithelial cytoplasms from the SS to stratum granulosum (SG), whereas caspase-8 was not detected in the epidermis. XIAP and Bak were found in the cytoplasm from the SS to SG, and the intensity of Bak-positivity was stronger in the SG than the SS, whereas Bid, Apaf-1 and cleaved caspase-9 were restricted in the SG. Homogenous cytoplasmic immunoreactivity of Bcl-2 was found in the SB and the intensity was gradually decreased from the SB to the SG. The granular-cytoplasmic immunopositivity of cytochrome C gradually altered into homogenous cytoplasmic expression in the upper half of the SG. Single-stranded DNA was rarely detected in the upper portion of the SG. These results suggest that epidermal apoptosis is induced by the interaction between Fas and Fas-L and the activation of caspase 10, and might initially proceed through a mitochondrial-independent pathway, and that a mitochondrial-dependent pathway finally accelerated under physiological conditions. PMID- 21630212 TI - Comparative immunohistochemical study of tissue integration of macroporous and laminar surgical meshes. AB - Intraperitoneal surgical mesh implantation is required for laparoscopic ventral hernia repair. Composite meshes are well known in animal models and human practice. The aim of our study is to compare the biological behaviour of two different textured silicone-covered polypropylene meshes. Transmural abdominal wall defect was created in 40 rabbits and treated as follows: In 20 animals a polypropylene mesh with a laminar silicone covering (LSPP) and in the rest a macroporous textured mesh knitted of silicone-impregnated polypropylene filaments (MSPP) was applied. One and three weeks after implantation we evaluated the intraperitoneal adhesion formation of the mesh macroscopically, histologically and immunohistochemically to detect the reactive cells, especially inflammatory, endothelial and mesothelial cells, as well as their proliferative activity, and with Scanning Electron microscopy to visualize the surface of the meshes. The adhesion formation caused by the composites showed no statistical difference after one week although in the three weeks old samples the LSPP adhesion was significantly weaker than that of MSPP. As complications, serome formation in both groups, fistulas, abscesses, and sc. haematoma in the LSPP group were found. Only in MSPP containing tissues was the decrease of Ki-67 positive proliferating cells significant. A significant increase in VEGF expressing cells was observed only in MSPP containing three week old samples, suggesting better regulation of vascular growth in tissues surrounding the implants. In one week old specimens we observed an irregular proliferation of cytokeratin containing mesothelial cells in both group. The intraperitoneal surface of MSPP mesh was covered with neoperitoneum, while it was not regularly seen on LSPP mesh after three week. PMID- 21630213 TI - Different patterns of apoptosis in response to cisplatin in B50 neuroblastoma rat cells. AB - Cisplatin (cisPt) is a chemotherapeutic drug used for several human malignancies. CisPt cytotoxicity is primarily mediated by its ability to cause DNA damage and subsequent apoptotic cell death. DNA is the primary target of cisPt; however, recent data have shown that cisPt may have important direct interactions with mitochondria, which can induce apoptosis and may account for a significant part of the clinical activity associated with this drug. We have previously demonstrated that in the rat neuronal cell line B50, at 20 h-treatment with cisPt activates apoptosis through an intrinsic pathway involving an alteration of mitochondrial membrane permeability and the release of cytochrome c. The present study investigates different death pathways induced in the same cell line by a prolonged treatment with 40 uM cisPt for 48 h. To address this issue, we focused on caspases-8 and -12, and on the mitochondrial apoptosis inducing factor (AIF), which translocates to the nucleus and induces cell death via caspase-independent pathway. We found that cisPt activates different forms of cell death, i.e. the receptor-mediated apoptotic extrinsic pathway and a death process mediated by endoplasmic reticulum stress. Moreover, we demonstrated that AIF-mediated death occurs, being characterized by the translocation of AIF from mitochondria to the nucleus. On the whole, we provided evidence that prolonged cisPt treatment is able to activate both caspase-dependent and caspase-independent apoptotic pathways in B50 rat neuronal cells. PMID- 21630214 TI - C-reactive protein and embolization during carotid artery stenting. A serological and morphological study. AB - INTRODUCTION: High-sensitivity C-Reactive Protein (hsCRP) levels are correlated with vulnerable carotid plaques, although their impact on the outcome of carotid revascularization is unknown. The aim of our study was to investigate the correlation between hsCRP and embolization during carotid artery stenting (CAS). METHODS: Patients with symptomatic carotid stenosis were submitted to CAS with distal protection filters. Serum hsCRP was analysed prior to CAS and patients were divided into two groups: Class I, patients presenting hsCRP < 5 mg/l and, Class II, patients presenting hsCRP>=5 mg/l. Plaques were categorised by ultrasound grey scale measurement as homogenous and dishomogenous. Afterwards CAS filters were analyzed microscopically and ultrastructurally to determine the type and the amount of debris present, based on percentage of surface involvement (SI) and pore occluded (PO) by embolic material. RESULTS: Fourteen patients underwent uneventful CAS, with a mean hsCRP of 11.5+/-18.4 mg/l. Eight patients were in Class I and six in Class II. All filters had microscopic debris. SI was 25.4% in Class I and 33.3% in Class II (p=ns), PO 22.9% and 33.3% respectively (p=0.049). Patients in Class II who also had a dishomogenous plaque showed greater SI and PO compared with patients in Class I with homogenous plaque (35.0% vs. 21.8% and 40.4% vs. 22.7% respectively, p<0.05). Microscopically embolic material was identified as atherosclerotic plaque fragments and platelet aggregates and was similar in both groups. DISCUSSION: High hsCRP levels are associated with significantly greater embolization during CAS in symptomatic patients, particularly in dishomogenous plaque. Although these results need further investigation due to the limited number of enrolled patients, this study suggests that CAS may not be indicated as a method of carotid revascularization in this setting. PMID- 21630215 TI - Significance of alpha-SMA in myofibroblasts emerging in renal tubulointerstitial fibrosis. AB - Myofibroblast transdifferentiation plays a crucial role in the development and progression of renal tubulointerstitial fibrosis. However, the significance of alpha-smooth muscle actin (alpha-SMA) expression, which is the major morphological characteristic of myofibroblasts, remains to be determined in detail. The effect of alpha-SMA expression on fibrosis tissue was examined by using a fibrosis model (collagen gel) in vitro. The transdifferentiation of fibroblasts into myofibroblasts was triggered in the culture medium with 0.5% fetal bovine serum (FBS)+transforming growth factor (TGF)-beta1, but not with 10% FBS+TGF-beta1. The TGF-beta1-induced gel contraction caused by myofibroblasts was greater than that by fibroblasts. Gel contraction by myofibroblasts involved the Ca2+-dependent myosin light chain kinase pathway, as well as the activation of Rho kinase and p38 mitogen-activated protein kinase (MAPK). Taken together, these findings suggest that alpha-SMA expression in renal interstitial fibroblasts, i.e., myofibroblast transdifferentiation, accelerates the contraction of the tubulointerstitial fibrosis tissue via the Ca2+-dependent pathway, in addition to the pathways involved in fibroblast contraction; this event may lead to renal atrophy and renal failure. PMID- 21630216 TI - Prospective evaluation of the learning curve of confocal laser endomicroscopy in patients with IBD. AB - BACKGROUND AND AIMS: Confocal laser endomicroscopy (CLE) represents a novel endoscopic imaging technique which enables the in vivo microscopic imaging within the mucosal layer of the gut at subcellular resolution. Currently, there are no data available on the learning curve of CLE, which was therefore the aim of our study. METHODS: Twenty-six consecutive patients with inflammatory bowel disease (IBD) underwent total colonoscopy and were examined by fluorescein-aided CLE. Image data were collected and reviewed by two endoscopists in a blinded fashion. CLE images were compared to endoscopical and histological findings. Prospectively, the following performance parameters were documented: total duration of the procedure, confocal imaging time, time to receive a confocal image in focus, number of confocal images, number of confocal images in focus, CLE diagnosis and final histopathological diagnosis. RESULTS: A significance decrease of CLE duration was detected between the first 8 and the subsequent cases (p=0.002). Confocal imaging time and the time to receive an image in focus declined significantly over time (p=0.0001), while number of images in focus significantly increased (p=0.0007). Agreement between CLE and histopathology improved over time with kappa values of 0.81 after twenty-six cases. CONCLUSIONS: There was a significant improvement in CLE performance over time, including decreased confocal imaging time, successful CLE diagnosis and decline in procedural time. These parameters improved significantly after the initial three cases. Therefore, CLE represents an easy to learn and apply novel diagnostic method for in vivo analysis and diagnosis in IBD. PMID- 21630217 TI - Biological effects of low-frequency pulsed magnetic fields on the embryonic central nervous system development. A histological and histochemical study. AB - Numerous experiments have yielded contradictory results on the harmful action of magnetic fields on embryonic development. Pulsed magnetic fields appear to be able to delay normal development of embryos. In the present study, fertilized Gallus domesticus eggs were exposed during incubation to pulsed magnetic fields (harmonic signals of 10 uT for 1 second with silences of 0.5 seconds) of 50 or 100 Hz frequency. Embryos extracted at 45 h of exposure to fields of 50 Hz or 100 Hz frequency had significantly (p<0.05) fewer somite pairs compared with controls of the same age. At 15 days of incubation, only embryos exposed to a 10 uT- 50 Hz field had a significantly (p<0.05) higher somatic weight. At 21 days of incubation, a significantly lower somatic weight (p<0.01) and development stage (p<0.05) was found in embryos exposed to a 10 uT-100 Hz field than in controls, while a lower development stage (p<0.05) alone was observed in those exposed to a 10 uT-50 Hz field. In addition, animals showed higher expression of the neural marker NSE (neural specific enolase) after 21 days of development as determined by immunohistochemistry, with very low expression of glycosaminoglycans identified by alcyan blue staining. These results suggest that pulsed magnetic fields may be able to hinder normal embryonic development in vivo and to alter normal neural function, at least at the intensities and frequencies analyzed in the present study. PMID- 21630218 TI - Histopathological changes in gerbil liver and kidney after aluminum subchronic intoxication. AB - Young gerbil livers and kidneys were analyzed by means of light and electron microscope to assess the histopathological changes caused by prolonged systemic aluminum (Al) administration. The experimental group was injected with AlCl3 i.p. for 5 weeks, while litter mates received PBS as sham-injected controls or served as untouched controls. Mortality occurred in 33% of experimental and 12.5% of sham-injected groups. The animals were perfused intracardially with 1% glutaraldehyde plus 1% paraformaldehyde and samples of liver and kidneys were processed for aluminum and iron histochemistry and conventional light- and transmission electron microscopy. White deposits composed of cellular debris appeared on the surface of liver and kidneys and in the mesentery as a consequence of Al treatment. Adherences of Glisson capsule to the diaphragm, as well as scattered small foci of hepatocyte necrosis with non-caseificant microgranulomas and mild portal inflammation, developed in the experimental group. Sham-injected animals also exhibited these granulomas but to a lesser degree. Al deposits were found in experimental animal granulomas and inside macrophages cytoplasm scattered throughout the liver. Iron deposition appeared in pericentral hepatocytes of experimental animals, in granulomas and in portal spaces of the three groups of animals. Ultrastructurally, hepatocytes of experimental animals showed mitochondria hyalinization, disintegration of endoplasmic reticulum and clustering of ribosomes. Phagolysosomes appeared larger and occurred more frequently in both hepatocytes and Kupffer cells of experimental animals. In 2 out of the 6 experimental animals studied, tubular atrophy was present in the renal cortical region, the kidneys of the remaining animals appearing normal. Al and iron were found very occasionally in the kidney parenchyma of experimental animals, while isolated mesangial cells showed iron deposits in a few glomeruli of both experimental and the two groups of control animals. PMID- 21630219 TI - Expression and regulation of nicotine receptor and osteopontin isoforms in human pancreatic ductal adenocarcinoma. AB - Osteopontin (OPN) is a secreted phospho-protein that confers on cancer cells a migratory phenotype. We have recently shown that nicotine, a risk factor in pancreatic ductal adenocarcinoma (PDA), induces an alpha7-nicotine acetylcholine receptor (alpha7-nAChR)-mediated increase of OPN in PDA cells. In this study, we tested nicotine's effect on the expression of OPN splice variants (OPNa, b, c) in PDA cells. We also analyzed the correlation between patients' smoking history with OPN and alpha7-nAChR levels. RT-PCR and UV-light-illumination of ethidium bromide staining were used to examine the mRNA expression in tissue and PDA cells treated with or without nicotine (3-300 nM). Localization of total OPN, OPNc and alpha7-nAChR was analyzed by immunohistochemistry, and their mRNA tissue expression levels were correlated with the patients' smoking history. PDA cells expressed varying levels of OPNa, OPNb, and alpha7-nAChR. Nicotine treatment selectively induced denovo expression of OPNc and increased alpha7-nAChR expression levels. In PDA tissue, OPNc was found in 87% of lesions, of which 73% were smokers. OPNc and total OPN levels were correlated in the tissue from patients with invasive PDA. Nicotine receptor was expressed in the invasive and premalignant lesions without clear correlation with smoking history. We show here for the first time that alpha7-nAChR is expressed in PDA cells and tissues and is regulated by nicotine in PDA cells. This, together with our previous findings that alpha7-nAChR mediates the metastatic effects of nicotine in PDA, suggest that combined targeting of alpha7-nAChR and OPNc could be a valid novel therapeutic strategy for invasive PDA, especially in the smoking population. PMID- 21630220 TI - beta-lapachone accelerates the recovery of burn-wound skin. AB - beta-lapachone is a quinone of lapachol extracted from the bark of lapacho tree. Recent findings demonstrated that punched skin wounds of mice healed faster with beta-lapachone treatment. The present study investigates the effects of beta lapachone on burn-wound skin of C57BL/6 mice injured by a 100 degrees C iron stick. Our results indicated that wounds treated with beta-lapachone recovered faster than those treated with control ointment containing no beta-lapachone. On the third day after burning, the area of beta-lapachone treated-wound was 30% smaller than wound treated with control ointment. H&E and immunohistochemistry staining showed that burn-wound skin treated with ointment containing beta lapachone healed faster in its epidermis, dermis, and underlying connective tissues with more macrophages appeared than those treated with control ointment alone. RAW264.7 cell, a macrophage-like cell line derived from BALB/C mice, was used as a model for scrutinizing the effect of beta-lapachone on macrophages. We found that the proliferation and the secretion of EGF and VEGF by macrophages were higher in cultures treated with beta-lapachone and that beta-lapachone can also increase the release of EGF with TNF-alpha pretreatment. We conclude that beta-lapachone plays an important role in accelerating burn wound healing, and that beta-lapachone not only can raise the proliferation of macrophages but also increase the release of VEGF from macrophages. PMID- 21630221 TI - The function of TRPS1 in the development and differentiation of bone, kidney, and hair follicles. AB - TRPS1 is a gene involved in Tricho-rhino-phalangeal syndrome (TRPS), an autosomal dominant skeletal disorder. TRPS1 encodes a GATA-type transcription factor that has nine zinc-finger motifs. A variety of mutations in TRPS1 including deletions and insertions, have been found in patients with TRPS type I and III. The functions of each domain of TRPS1 have been clarified from study of these mutations. Further studies on the localization and the function of TRPS1 have been performed using TRPS1Deltagt and Trps1-deficient mice, which allow examination of the development and differentiation of all tissues with Trps1 expression. These studies suggest that TRPS1 exhibits a variety of functions in cartilage, kidneys, and hair follicles. In the growth plate cartilage, TRPS1 regulates the differentiation, proliferation, and apoptosis of chondrocytes through interaction of several signaling molecules. In addition, TRPS1 has a function downstream of BMP7, which regulates the mesenchymal-epithelial transition when nephrons are formed in renal development. Furthermore, TRPS1 suppresses the epithelial-mesenchymal transition and renal fibrosis induced by unilateral ureteral obstruction by decreasing Arkadia expression. Finally, TRPS1 is expressed in the dermal papillae and the mesenchymal cells surrounding the hair pegs, and the loss of TRPS1 largely influences the development of hair follicles. The molecular mechanisms of the function of TRPS1 in cartilage, kidneys, and hair follicles are discussed in this review. PMID- 21630222 TI - Cell- and gene-therapy approaches to inner ear repair. AB - Sensorineural hearing loss is the most common sensory disorder in humans. It is primarily due to the degeneration of highly specialised mechanosensory cells in the cochlea, the so-called hair cells. Hearing problems can also be caused or further aggravated by the death of auditory sensory neurons that convey the information from the hair cells to the brain stem. Despite the discovery of stem/progenitor cells in the mammalian cochlea, no regeneration of either damaged hair cells or auditory neurons has been observed in mammals, in contrast to what is seen in avians and non-mammalian vertebrates. The reasons for this divergence have not yet been elucidated, although loss of stem cells and/or loss of their phenotypic plasticity in adult mammals have been put forward as possible explanations. Given the high incidence of this disorder and its economic and social implications, a considerable number of research lines have been set up aimed towards the regeneration of cochlear sensory cell types. This review summarizes the various routes that have been explored, ranging from the genetic modification of endogenous cells remaining in the inner ear in order to promote their transdifferentiation, to the implantation of exogenous stem or progenitor cells and their subsequent differentiation within the host tissue. Prophylactic treatments to fight against progressive sensory cell degeneration in the inner ear are also discussed. PMID- 21630223 TI - Immunosuppressive cells and tumour microenvironment: focus on mesenchymal stem cells and myeloid derived suppressor cells. AB - Tumours have been compared to unhealed wounds that produce large amounts of inflammatory mediators, including cytokines, chemokines, and growth factors. These molecules participate in the formation of a rich and heterogeneous microenvironment by attracting non malignant cells that promote tumour progression and dissemination. Tumour infiltrating cells include macrophages, myeloid-derived suppressor cells (MDSCs), mesenchymal stromal cells (MSCs) and TIE2-expressing monocytes. Most of them are bone marrow-derived, although MSC are present in virtually every tissue. This review focuses on MDSCs and MSCs, both of which can exert pro-tumorigenic effects through negative regulation of immune responses. MDSCs represent a heterogeneous population of cells of myeloid origin that are expanded and activated in response to growth factors and cytokines released by tumours. Once MDSCs are activated, they accumulate in lymphoid organs and tumours where they exert T cell immunosuppression. Like MDSCs, MSCs can be mobilized from the bone marrow into the bloodstream and home in the tumour stroma, where they either help or hinder tumour growth. Here, we will discuss the origin, the functions and the mechanisms of action of MSCs and MDSCs, as well as the strategies to target these cells for the therapeutic benefit of cancer patients. PMID- 21630224 TI - Human ligaments classification: a new proposal. AB - A high concern exists among physicians about surgically important ligaments such as cruciate and collateral ligaments of the knee, patellar ligament, tibiofibular syndesmosis, collateral ligaments of the ankle, and coracoclavicular ligament. However, the classification of the ligaments is insufficient in the literature, due to their origin from connective tissue. A new classification is proposed, based on various parameters such as the macroscopic and microscopic features, the function and the nature of their attachment areas. PMID- 21630225 TI - A bicarotid trunk in association with an aberrant right subclavian artery. Report of two cases, clinical impact, and review of the literature. AB - A combination of an aberrant right subclavian artery (ARSA) and a bicarotid trunk (BCT) appears in up to 2.5% of the population. The aim of this study is to report the higher total and male incidence of this variation in the literature and to summarise its clinical impact, providing useful knowledge to anatomists, radiologists, cardiologists, and vascular and thoracic surgeons in order to avoid diagnostic pitfalls and therapeutic complications. A total of 72 (43 female and 29 male) Greek Caucasian formalin-embalmed cadavers were studied. The international literature was reviewed along with the dissection archives of the Department of Anatomy from 1986 to 2009. Two male cadavers were found to have an ARSA combined with a BCT (incidence: total 2.78%, males 6.9%, females 0%). Both aortic arches consisted of three branches: (1) the BCT, (2) the left subclavian artery, and (3) the ARSA. The common carotids followed a normal route to the neck; the ARSA passed between the trachea and the oesophagus in the first case and behind the oesophagus in the second case, and was accompanied by a non recurrent laryngeal nerve. In the second cadaver the ARSA formed a sharp angle (kinking/buckling) on its route to the right arm. The ARSA is associated with several congenital cardiovascular anomalies and some chromosomal and other syndromes. It is occasionaly responsible for causing dysphagia, dyspnoea, or acute ischaemia to the right upper limb, and it may present as a superior mediastinal mass in cases of aneurysm formation. PMID- 21630226 TI - Histological and ultrastructural studies of the olfactory epithelium of spotted butter fish Scatophagus argus (Linnaeus). AB - The olfactory epithelium of Scatophagus argus (Linnaeus) was investigated by light and scanning electron microscopy. The elongated olfactory organ is made up of 20 to 22 primary lamellae arranged on both sides of the narrow median raphe. Sensory and non-sensory regions are located separately on each lamella. The sensory epithelium occupies the upper apical broad half and extreme basal part of the olfactory lamellae whereas the middle slender part is covered with non sensory epithelium. The sensory epithelium consists of ciliated, microvillus, and crypt cells. The non-sensory epithelium is made up of stratified epithelial cells having different patterns of finger-like micro-ridges and mucous cells. Different cells lining the olfactory epithelium have been correlated with the functional views of the fish concerned. PMID- 21630227 TI - Basilar artery fenestration. AB - The posterior circulation of the brain constitutes the vertebrobasilar system and its branches, which are responsible for about 30% of the brain's blood supply. The aim of this study was to describe the anomalies of the basilar artery, especially fenestrations. For that purpose, we examined 50 patients with computed tomography (CT) angiography during an 8-month period. In the CT reports of 2 (4%) patients of the 50 analysed, fenestration was found at the proximal basilar trunk. The two fenestrations in our series were not associated with aneurysms. No collateral branches originated from the two limbs of the fenestration. In conclusion, basilar artery fenestrations are a rare finding. The data derived from this study are useful teaching material for anatomists, and for the radiologists and neurosurgeons they are important for diagnostic and intervention procedures such as CT, magnetic resonance imaging, angiography, and surgical and endovascular procedures. PMID- 21630228 TI - Median aperture of the fourth ventricle revisited. AB - BACKGROUND: The median aperture of Magendie is the largest of three openings of the fourth ventricle and thus it forms the main path for the outflow of the cerebrospinal fluid from the ventricle. The Magendie aperture connects the fourth ventricle with the cisterna magna and makes a natural corridor for neurosurgical approach and inspection of the ventricle and its floor. The purpose of this study was to give a contemporary anatomical view of this structure in the context of historical data. MATERIAL AND METHODS: The Magendie foramen was studied in 30 fixed specimens of human brainstems with cerebella. The microdissection technique was used. Measurements were taken with a microscope ocular ruler. RESULTS: The aperture is limited by the following structures: obex and gracile tubercles inferiorly, and tela choroidea with choroid plexus superolaterally. Obex tubercles usually have the form of a piece of neural tissue bridging two halves of the brainstem above the entrance to the central canal. Gracile tubercles together are 8.15 mm wide and the maximal width of the foramen is 6.53 mm. Tela choroidea attaches laterally at both sides to the inferior medullary velum. In most cases the right and left choroid plexus are connected to each other with a triangular membrane of tela choroidea, which protrudes through the median foramen and attaches to the vermis at a highly variable level. CONCLUSIONS: We hope that the presented description of anatomical relations around the Magendie aperture, with its new measurements, will be helpful for those operating in the area and will explain some of the inaccuracies found in literature. PMID- 21630229 TI - Lateral approach to the ankle and distal leg. AB - Fractures of the distal tibia are usually high-energy injuries and are well known for their soft tissue complications after surgery. Various factors have been studied regarding the possibility of them reducing the incidence of such damage, including timing of surgery, staged surgery, fixation method, and surgical approach. Less invasive surgical techniques with vascularity preserving approaches were proposed as reasonable solutions to this problem. The aim of this study was to investigate the presence of minor vascular structures, which can be preserved during anterolateral approach, possibly contributing to the success of the approach. Lateral approach for the distal tibia was performed in 22 cadaver feet. The mean distance between the lateral malleolus and the superficial peroneal nerve was 12.2 cm. Two different vascular pedicles, from peroneal vessels to muscles of the anterior compartment, were 4.3 and 8.2 cm away from the lateral malleolus, respectively. We conclude that preserving greater vascularity was possible in the lateral approach for the distal tibia, placing the plate in a completely submuscular plane. PMID- 21630230 TI - Assessment of morphological and functional changes in neonate vitrified testis grafts after host treatment with melatonin. AB - This study was conducted to assess the effect of melatonin on the structure of testis and spermatogenesis dynamics in neonate vitrified testis grafts. Neonate vitrified testes, candidates for transplantation heterotopically to experiment or control groups, were warmed in thawing media which had or did not have a supplement of 100 MUM melatonin, respectively. Following transplantation, melatonin (20 mg/ /kg/day) or saline solution was intraperitoneally injected into the treated and the non-treated groups, respectively. The initiating spermatogenesis, spermatogonia survival, and structure of tissue in the testis graft were examined. Cell apoptosis (TUNEL assay) and proliferation (Brdu assay) in germ cells were determined. Histological studies revealed the dynamic of the spermatogenesis process in the vitrified testis graft. However, dilation of the lumen accompanied by a disorganised epithelium in the non-treated group was higher than in the treated group. Furthermore, the proportion of apoptotic germ cells together with a reduced proportion of proliferated germ cells was higher in the non-treated group than in the treated group. Overall, the number of seminiferous tubules in the testes grafts of both groups remained steady. However, the non-treated testes grafts contained more damaged seminiferous tubules than the treated ones. The thickness of the seminiferous tubules was greater in the melatonin treated group than in the non-treated group. In fact, the thickness of germinal epithelium was significantly higher in the treated group than in the non-treated group. The study may show a positive effect from melatonin resulting in more grafts restoring puberty. Furthermore, the associated increase in the healthy number of seminiferous tubules suggests that melatonin may have a preventative ischaemia/antioxidant role and in fact may be useful to initiate the spermatogenesis process. PMID- 21630231 TI - Morphometric features of the thyroid gland: a cadaveric study of Turkish people. AB - BACKGROUND: Although racial and ethnic variations in the morphology of anatomical structures are defined well, the size, shape, and weight of the thyroid gland have not previously been reported in Turkish people. This study provides data about the morphometric features of the thyroid gland, thyroid lobes, and pyramidal lobe, and highlights some anatomical variations in people from the Marmara region in Turkey. MATERIAL AND METHODS: The material for the present study consisted of thyroid glands obtained from 75 male and 15 female adult cadavers aged between 18 and 80 years. A dissection was carried out and the thyroid glands were exposed. The glands were weighed and measured according to the various age groups of the patients. RESULTS: A pyramidal lobe was found to be present in 57.8% of the cadavers (52/90). During midline dissection of the neck 2 males out of 90 cadavers, giving an incidence of 2.22%, did not show an isthmus. The mean thyroid weight was 26.11 +/- 8.14 g. In males it was 26.93 +/- 7.96 g while in females it was 21.93 +/- 7.98 g. CONCLUSIONS: This is the first reported morphometric study on cadaveric thyroid glands from Turkey and it highlights individual and ethnic/racial variations. In order to perform safe and effective surgery and for the accurate diagnosis of thyroid disorders, knowledge of normal anatomy and the variations of the thyroid gland are essential. PMID- 21630232 TI - Correlation between morphometry of the suprascapular notch and anthropometric measurements of the scapula. AB - The concept of the study was to find the correlation between the morphometry of the suprascapular notch and basic anthropometric measurements of the human scapula. The measurements of the human scapulae included: morphological length and width, maximal width and length projection of scapular spine, length of acromion, and maximal length of the coracoid process. The glenoid cavity was measured in two perpendicular directions to evaluate its width and length. The width-length scapular and glenoid cavity indexes were calculated for every bone. In addition to standard anthropometric measurements two other measurements were defined and evaluated for every suprascapular notch: maximal depth (MD) and superior transverse diameter (STD). The superior transverse suprascapular ligament was completely ossified in 7% of cases. Ten (11.6%) scapulae had a discrete notch. In the studied material, in 21 (24.4%) scapulae the MD was longer than the STD. Two (2.3%) scapulae had equal maximal depth and superior transverse diameter. In 47 (57.7%) scapulae the superior transverse diameter was longer than the maximal depth. There was no statistically significant difference between anthropometric measurements in the group with higher MD and the group with higher STD. The maximal depth of the suprascapular notch negatively correlated with the scapular width-length index. The maximal depth of the scapular notch correlated with the morphological length of the scapulae. PMID- 21630233 TI - An anatomical study on the three-headed biceps brachii in human foetuses, and clinical relevance. AB - The biceps brachii (BB) is as one of the most variable muscles in the human body in terms of number and morphology of its heads. The most frequent variation is the presence of a third head, which has been reported by several authors in different populations. Our aim was to find the occurrence of the supernumerary head of BB in Turkish foetuses. Out of the 24 upper limbs of the foetuses, two (8.33%) arms were found to have a three-headed BB. The variations were present unilaterally in the right arm of one male foetus and one female foetus. In one of the cases, the third head of BB originated from the anteromedial aspect of the humerus just distal to the insertion of the coracobrachilais, medial to the brachialis, and in the other the third head was a thin muscle bundle, which arose on the lateral side of the insertion of coracobrachialis and over the origin of the brachialis. Both of them were extended distally and joined the common tendon. The occasional presence of the three-headed BB in the foetuses observed in the present study was similar to those of adults reported in previous studies. In conclusion, these variations are not rare and are interesting not only to anatomists but also to orthopaedic surgeons, plastic surgeons, traumatologists, physiotherapists, doctors dealing with sports medicine, and radiologists. PMID- 21630234 TI - Muscle pathology in myotonic dystrophy: light and electron microscopic investigation in eighteen patients. AB - Myotonic dystrophy (DM) is the most common muscular dystrophy in adults. Two known genetic subtypes include DM1 (myotonic dystrophy type 1) and DM2 (myotonic dystrophy type 2). Genetic testing is considered as the only reliable diagnostic criterion in myotonic dystrophies. Relatively little is known about DM1 and DM2 myopathology. Thus, the aim of our study was to characterise light and electron microscopic features of DM1 and DM2 in patients with genetically proven types of the disease. We studied 3 DM1 cases and 15 DM2 cases from which muscle biopsies were taken for diagnostic purposes during the period from 1973 to 2006, before genetic testing became available at our hospital. The DM1 group included 3 males (age at biopsy 15-19). The DM2 group included 15 patients (5 men and 10 women, age at biopsy 26-60). The preferential type 1 fibre atrophy was seen in all three DM1 cases in light microscopy, and substantial central nucleation was present in two biopsies. Electron microscopy revealed central nuclei in all three examined muscle biopsies. No other structural or degenerative changes were detected, probably due to the young age of our patients. Central nucleation, prevalence of type 2 muscle fibres, and the presence of pyknotic nuclear clumps were observed in DM2 patients in light microscopy. Among the ultrastructural abnormalities observed in our DM2 group, the presence of internal nuclei, severely atrophied muscle fibres, and lipofuscin accumulation were consistent findings. In addition, a variety of ultrastructural abnormalities were identified by us in DM2. It appears that no single ultrastructural abnormality is characteristic for the DM2 muscle pathology. It seems, however, that certain constellations of morphological changes might be indicative of certain types of myotonic dystrophy. PMID- 21630235 TI - Variations in hepatic vascularisation: lack of a proper hepatic artery. Two case reports. AB - The blood supply of the liver and other abdominal organs plays a significant role during abdominal surgery. Knowledge of the most common patterns of vascularisation should be broadened and new anomalies of the celiac trunk and its branches dutifully reported. This paper presents two case reports which describe the lack of a proper hepatic artery. Case 1 describes the cadaver of a 64-year old female in whom the right hepatic artery was observed to arise from the common hepatic artery and run behind the portal vein. The common hepatic artery was observed to be divided into three terminal vessels: the left hepatic artery, the gastroduodenal artery, and the right gastric artery. Case 2 describes the cadaver of a 75-year-old male with a liver that was supplied from 3 different sources: the left hepatic artery from the left gastric artery (which arose directly from the aorta), the right hepatic artery from the superior mesenteric artery, and the middle hepatic artery from the common hepatic artery - (branch of the hepato splenic trunk). Moreover, the left inferior phrenic artery arose from the left hepatic artery. PMID- 21630236 TI - Cor triloculare biventriculare with left superior vena cava. AB - Cor triloculare biventriculare is a rare congenital malformation of the heart in which there is a complete absence of the atrial septum. It is usually associated with other anomalies like complete atrioventricular canal defect, polysplenic syndrome, isolated dextrocardia, Ellis-van Creveld syndrome, or persistent left superior vena cava. We report a case of a stillborn male foetus of 35 weeks gestation with common atrium, complete atrioventricular canal defect, and persistent left superior vena cava. The possible embryological basis and clinical implication of this variation are discussed. PMID- 21630237 TI - Professor Olgierd Narkiewicz--the great Polish anatomist and neuroanatomist of the twentieth century (1925-2010). PMID- 21630240 TI - Rheumatoid arthritis and pregnancy: beyond smaller and preterm babies. PMID- 21630241 TI - Sulfasalazine for the treatment of ankylosing spondylitis: relic or niche medication? PMID- 21630242 TI - Closing ranks: the chondrocytes' move in bad times or biomechanical job sharing? PMID- 21630243 TI - Pregnancy and delivery in women with chronic inflammatory arthritides with a specific focus on first birth. AB - OBJECTIVE: To examine possible associations between chronic inflammatory arthritides and pregnancy outcomes with separate analyses of first and subsequent births before and after diagnosis. METHODS: Linkage of data from a registry of patients with chronic inflammatory arthritides and the Medical Birth Registry of Norway enabled a comparison of pregnancy outcomes in women with chronic inflammatory arthritides and pregnancy outcomes in reference subjects. Outcomes of first birth and subsequent births before and after diagnosis were analyzed separately. Associations between chronic inflammatory arthritides and the women's health during pregnancy and delivery as well as perinatal outcomes were assessed in logistic regression analyses with adjustments for maternal age at delivery and gestational age. RESULTS: We analyzed 128 first births and 151 subsequent births after diagnosis and 286 first births and 262 subsequent births before diagnosis in patients and compared them with first and subsequent births in reference subjects. Firstborn children of women diagnosed as having chronic inflammatory arthritides were more often preterm (odds ratio [OR] 1.85 [95% confidence interval (95% CI) 1.09-3.13]) and small for gestational age (OR 1.60 [95% CI 1.00 2.56]). They also had lower mean birth weight (P=0.01) and higher perinatal mortality (OR 3.26 [95% CI 1.04-10.24]). Birth by caesarean section (all classifications) was more frequent in patients than in reference subjects, and elective caesarean section was 2-fold more frequent in patients, both in first birth (OR 2.60 [95% CI 1.43-4.75]) and in subsequent births (OR 2.18 [95% CI 1.33 3.58]). No excess risks of clinical importance were observed prior to diagnosis of chronic inflammatory arthritides. CONCLUSION: Excess risks were related to first birth in women diagnosed as having chronic inflammatory arthritides, including a higher rate of perinatal mortality. A higher caesarean section rate was related to all patient deliveries. Mainly, pregnancy outcomes before diagnosis did not differ from those in reference subjects. PMID- 21630244 TI - Clinical images: Bone sarcoidosis mimicking multiple metastases. PMID- 21630245 TI - Clinical efficacy and safety of etanercept versus sulfasalazine in patients with ankylosing spondylitis: a randomized, double-blind trial. AB - OBJECTIVE: Etanercept, a fully human tumor necrosis factor (TNF) receptor, is an effective treatment in patients with ankylosing spondylitis (AS). Sulfasalazine is frequently used for the treatment of both axial symptoms and peripheral symptoms of AS, and it has been the recommended therapy before the use of an anti TNF agent when peripheral arthritis is present. Until now, no clinical trial has compared the efficacy and safety of a TNF blocker with that of sulfasalazine. This study was undertaken to compare the efficacy and safety of etanercept with that of sulfasalazine after 16 weeks of treatment in patients with axial and peripheral manifestations of AS. METHODS: In this randomized, double-blind study, patients received etanercept 50 mg once weekly (n=379) or sulfasalazine titrated to a maximum of 3 gm/day (n=187) for 16 weeks. The primary end point was the proportion of patients who achieved the Assessment of SpondyloArthritis international Society criteria for 20% improvement (ASAS20) at 16 weeks. Last observation carried forward was predefined for imputation of missing values. RESULTS: The mean age of the patients was 41 years, 74% were male, and the mean disease duration was 7.6 years. The proportion of ASAS20 responders at week 16 was greater among patients treated with etanercept compared with those treated with sulfasalazine (75.9% versus 52.9%; P<0.0001). As early as week 2 of treatment, etanercept was found to be more effective than sulfasalazine (P<0.0001) in ameliorating both the axial symptoms and peripheral manifestations. Serious adverse events rarely occurred, and the rate of serious adverse events did not differ between groups. CONCLUSION: In this population of patients with AS, etanercept was significantly more effective than sulfasalazine in improving the signs and symptoms of AS in the axial skeleton and peripheral joints. PMID- 21630246 TI - Onset of preclinical osteoarthritis: the angular spatial organization permits early diagnosis. AB - OBJECTIVE: Superficial articular chondrocytes display distinct spatial remodeling processes in response to the onset of distant osteoarthritis (OA). Such processes may be used to diagnose early events before manifest OA results in tissue destruction and clinical symptoms. Using a novel method of spatial quantification by calculating the angles between a chondrocyte and its surrounding neighbors, we compared maturational and degenerative changes of the cellular organizations in rat and human cartilage specimens. METHODS: The nuclei of superficial chondrocytes obtained from intact rat cartilage and from human knee cartilage, as well as from cartilage with focal and severe OA, were digitally recorded in top down views. Their Cartesian coordinates were used to determine the nearest neighbor for each chondrocyte and the angle between these 2 cells and a reference. These angles, cellularity, nearest neighbor distances, and aggregation were analyzed as a function of location and OA severity. RESULTS: Neighboring rat chondrocytes exhibited intricate angular patterns with 4 dominant angles that were maintained during maturation and during the onset and progression of OA. Within intact cartilage, human chondrocytes demonstrated 1 dominant angle and, thus, a significantly different angular organization. With early OA onset, human chondrocytes that were located within intact cartilage displayed an increased occurrence of 4 angles; the resulting angular patterns were indistinguishable from those observed in rats. The angular remodeling was associated with location- and OA severity-dependent changes in cellularity and aggregation. CONCLUSION: This study is the first to identify the presence of angular characteristics of spatial chondrocyte organization and species-specific remodeling processes correlating with OA onset. The appearance of distinct angular and spatial patterns between neighboring chondrocytes can identify the onset of distant OA prior to microscopically visible tissue damage and possibly before clinical onset. With further development, this novel concept may become suitable for the diagnosis and followup of patients susceptible to OA. PMID- 21630247 TI - Prenatal exposure to LPS leads to long-lasting physiological consequences in male offspring. AB - Growing evidence suggests that early life events are critical determinants for disorders later in life. According to a comprehensive number of epidemiological/animal studies, exposure to lipopolysaccharide, causes alteration in pro-inflammatory cytokine levels, hypothalamic-pituitary-adrenal functioning and the hormonal system which may contribute to behavioral and neurological injuries. In this study we investigated the effects of lipopolysaccharide administration on physiological parameters in pregnant dams and their male offspring aged 9 weeks. In gestational Day 10, pregnant mice were injected intrapritoneally with Salmonella enterica lipopolysaccharide to model prenatal exposure to infection. The following results were obtained for offspring from dams stressed during pregnancy: (a) reduced anxiety-related behavior in the elevated plus maze; (b) reduced food and water intake; (c) reduced body weight from birth up to postnatal Day 40. The observed data provide experimental evidence showing that prenatal stress can have complex and long-lasting physiological/behavioral consequences in offspring. PMID- 21630248 TI - Early viral replication in lymph nodes provides HIV with a means by which to escape NK-cell-mediated control. AB - Acute HIV infection is marked by dramatic viral replication associated with preferential replication within secondary lymphoid tissues, such as lymph nodes (LNs), that is rapidly but incompletely contained to a viral setpoint. Accumulating evidence supports a role for natural killer (NK) cells in the early control of HIV infection; however, little is known about the location of their antiviral control. Given that HIV replicates profusely in LNs during early infection, we sought to define whether changes occurred in the NK cell infiltrate within these sites during the first year of HIV infection. Surprisingly, NK cell numbers and distribution were unaltered during early HIV infection. LN NK cells expressed decreased inhibitory receptors, were more highly activated, and expressed elevated TRAIL, potentially conferring a superior capacity for NK cells to become activated and control infection. Most noticeably, KIR(+) NK cells were rarely detected in the LN during HIV infection, associated with diminished migratory capacity in the setting of reduced expression of CX3CR1 and CXCR1. Thus, incomplete control of HIV viral replication during early disease may be due to the inefficient recruitment of KIR(+) NK cells to this vulnerable site, providing HIV a niche where it can replicate unabated by early NK-cell-mediated innate pressure. PMID- 21630249 TI - Antigen-presentation capacity of dendritic cells is impaired in ongoing enterovirus myocarditis. AB - Coxsackievirus B3 (CVB3)-infection is a frequent cause of acute myocarditis, which may result in chronic myocarditis and virus persistence. Investigation of the initial immune responses to CVB3 may shed light on the mechanisms that contribute to ongoing disease. DCs, as key professional APCs, were investigated in two MHC-matched hosts: while C57BL/6 mice are resistant to chronic CVB3 myocarditis, the A.BY/SnJ mouse strain exhibits susceptibility. DC maturation and activation were critically impaired in A.BY/SnJ mice, as reflected by the failure of DCs to induce co-stimulatory molecules and cytokine/chemokine responses. MHC class I-restricted antigen presentation via the cross-presentation pathway was also affected in DCs from A.BY/SnJ mice. DC maturation involves the accumulation of DC aggresome-like induced structures (DALISs) and the transient storage of defective ribosomal products (DRiPs). DCs from A.BY/SnJ mice showed permanent DALIS accumulation; the detection of poly-ubiquitinylated CVB3 proteins in these DALISs suggested a limitation in the MHC class I antigenic supply in this host. In conclusion, ongoing chronic disease in A.BY/SnJ mice due to a failure to clear the virus may be attributed to defects in DC maturation/activation and DC MHC class I antigen processing. PMID- 21630250 TI - TLR activation enhances C5a-induced pro-inflammatory responses by negatively modulating the second C5a receptor, C5L2. AB - TLR and complement activation ensures efficient clearance of infection. Previous studies documented synergism between TLRs and the receptor for the pro inflammatory complement peptide C5a (C5aR/CD88), and regulation of TLR-induced pro-inflammatory responses by C5aR, suggesting crosstalk between TLRs and C5aR. However, it is unclear whether and how TLRs modulate C5a-induced pro-inflammatory responses. We demonstrate a marked positive modulatory effect of TLR activation on cell sensitivity to C5a in vitro and ex vivo and identify an underlying mechanistic target. Pre-exposure of PBMCs and whole blood to diverse TLR ligands or bacteria enhanced C5a-induced pro-inflammatory responses. This effect was not observed in TLR4 signalling-deficient mice. TLR-induced hypersensitivity to C5a did not result from C5aR upregulation or modulation of C5a-induced Ca(2+) mobilization. Rather, TLRs targeted another C5a receptor, C5L2 (acting as a negative modulator of C5aR), by reducing C5L2 activity. TLR-induced hypersensitivity to C5a was mimicked by blocking C5L2 and was not observed in C5L2KO mice. Furthermore, TLR activation inhibited C5L2 expression upon C5a stimulation. These findings identify a novel pathway of crosstalk within the innate immune system that amplifies innate host defense at the TLR-complement interface. Unravelling the mutually regulated activities of TLRs and complement may reveal new therapeutic avenues to control inflammation. PMID- 21630251 TI - Adaptive immune defects against glycoantigens in chronic granulomatous disease via dysregulated nitric oxide production. AB - Chronic granulomatous disease (CGD) is a primary immunodeficiency defined by mutations in the NADPH oxidase complex leading to reduced superoxide production, increased susceptibility to infection, chronic inflammation, and recurring abscess and granuloma formation. Here, we found that CGD mice were hyperresponsive to abscess-inducing T-cell-dependent carbohydrate antigens (glycoantigens) due to a ten-fold increase in NO production within APCs, which is known to be necessary for glycoantigen presentation on MHC class II. CGD mice exhibited increased Th1 pro-inflammatory T-cell responses in vitro and in vivo, characterized by more severe abscess pathology. This phenotype was also seen in WT animals following adoptive transfer of neutrophil-depleted APCs from CGD animals, demonstrating that this phenotype was independent of neutrophil and T cell defects. Finally, pharmacological attenuation of NO production to WT levels in vivo reduced abscess incidence and severity in CGD without overt increases in inflammation or the ability to clear infection, suggesting a potential new treatment option for early stage CGD-associated infections. PMID- 21630252 TI - T-zone localized monocyte-derived dendritic cells promote Th1 priming to Salmonella. AB - Control of intracellular Salmonella infection requires Th1 priming and IFN-gamma production. Here, we show that efficient Th1 priming after Salmonella infection requires CD11c(+) CD11b(hi) F4/80(+) monocyte-derived dendritic cells (moDCs). In non-infected spleens, moDCs are absent from T-cell zones (T zones) of secondary lymphoid tissues, but by 24 h post-infection moDCs are readily discernible in these sites. The accumulation of moDCs is more dependent upon bacterial viability than bacterial virulence. Kinetic studies showed that moDCs were necessary to prime but not sustain Th1 responses, while ex vivo studies showed that antigen experienced moDCs were sufficient to induce T-cell proliferation and IFN-gamma production via a TNF-alpha-dependent mechanism. Importantly, moDCs and cDCs when co-cultured induced superior Th1 differentiation than either subset alone, and this activity was independent of TNF-alpha. Thus, optimal Th1 development to Salmonella requires the rapid accumulation of moDCs within T zones and their collaboration with cDCs. PMID- 21630253 TI - The earliest intrathymic precursors of CD8alpha(+) thymic dendritic cells correspond to myeloid-type double-negative 1c cells. AB - The dendritic cells (DCs) present in lymphoid and non-lymphoid organs are generated from progenitors with myeloid-restricted potential. However, in the thymus a major subset of DCs expressing CD8alpha and langerin (CD207) appears to stand apart from all other DCs in that it is thought to derive from progenitors with lymphoid potential. Using mice expressing a fluorescent reporter and a diphtheria toxin receptor under the control of the cd207 gene, we demonstrated that CD207(+) CD8alpha(+) thymic DCs do not share a common origin with T cells but originate from intrathymic precursors that express markers that are normally present on all (CD11c(+) and MHCII molecules) or on some (CD207, CD135, CD8alpha, CX3CR1) DC subsets. Those intrathymic myeloid-type precursors correspond to CD44(+) CD25(-) double-negative 1c (DN1c) cells and are continuously renewed from bone marrow-derived canonical DC precursors. In conclusion, our results demonstrate that the earliest intrathymic precursors of CD8alpha(+) thymic DCs correspond to myeloid-type DN1c cells and support the view that under physiological conditions myeloid-restricted progenitors generate the whole constellation of DCs present in the body including the thymus. PMID- 21630254 TI - The synergistic therapeutic effect of cisplatin with Human papillomavirus E6/E7 short interfering RNA on cervical cancer cell lines in vitro and in vivo. AB - Human papillomavirus (HPV) types 16 and 18 are the major etiologic factors in the development of cervical epithelial neoplasia. Our study was designed to validate antiviral short interfering RNA (siRNA) targeting the E6 and E7 oncogenes as a potential chemosensitizer of cisplatin (cis-diaminedichloroplatinum II; CDDP) in cervical carcinoma. Specifically, the therapeutic efficacy of combination of CDDP and E6/E7-specific siRNA was assessed in an in vivo cervical cancer xenograft models. The combination of CDDP and E6/E7-specific siRNA had greater efficacy than the combination of CDDP and E6-specific siRNA especially in terms of inducing cellular senescence. Through in vitro and in vivo experiments, the mechanism of synergy between these two treatments was revealed, demonstrating that the combination of E6/E7-specific siRNA and CDDP therapy was significantly superior to either modality alone. In vitro, long-term exposure of HeLa cells to the combination of CDDP and E6/E7-specific siRNA induced apoptosis and cellular senescence. In vivo, E6/E7-specific siRNA potentiated the antitumor efficacy of CDDP via induction of apoptosis, senescence and antiangiogenesis. Our results suggest that E6/E7-specific siRNA may be an effective sensitizer of CDDP chemotherapy in cervical cancer. PMID- 21630255 TI - Improved sensitivity of vaginal self-collection and high-risk human papillomavirus testing. AB - Self-collected vaginal specimens tested for high-risk human papillomavirus (HR HPV) have been shown to be less sensitive for the detection of cervical intraepithelial neoplasia or cancer (>=CIN 3) than physician-collected endocervical specimens. To increase the sensitivity of self-collected specimens, we studied a self-sampling device designed to obtain a larger specimen from the upper vagina (POI/NIH self-sampler) and a more sensitive polymerase chain reaction (PCR)-based HR-HPV assay. Women (10,000) were screened with cervical cytology and HR-HPV testing of vaginal self-collected and endocervical physician collected specimens. Women were randomly assigned to use either a novel self collection device (POI/NIH self-sampler) or conical-shaped brush (Qiagen). The self-collected and clinician-collected specimens were assayed by Cervista (Hologic) and the research only PCR-based matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF). Women with any abnormal screening test underwent colposcopy and biopsy. Women (8,556), mean age of 38.9, had complete data; 1.6% had >= CIN 3. For either HR-HPV assay, the sensitivity was similar for the two self-collection devices. Tested with Cervista, the sensitivity for >=CIN 3 of self-collected specimens was 70.9% and for endocervical specimens was 95.0% (p = 0.0001). Tested with MALDI-TOF, the sensitivity for >=CIN 3 of self-collected specimens was 94.3% and for endocervical specimens was also 94.3% (p = 1.0). A self-collected sample using a PCR-based assay with the capability of very high throughput has similar sensitivity as a direct endocervical specimen obtained by a physician. Large population-based screening "events" in low-resource settings could be achieved by promoting self-collection and centralized high-throughput, low-cost testing by PCR-based MALDI-TOF. PMID- 21630256 TI - Antibodies reactive to Plasmodium falciparum serine repeat antigen in children with Burkitt lymphoma from Ghana. AB - The role of protective immunity to Plasmodium falciparum (Pf) malaria in Burkitt lymphoma (BL) is unknown. We investigated the association between BL and antibodies reactive to SE36 antigen, a recombinant protein based on P. falciparum serine repeat antigen 5 gene, targeted by protective malaria immune responses. Cases were children (0-14 years) enrolled at the Korle-Bu Teaching Hospital, Accra, Ghana, during 1965-1994 with BL confirmed by histology or cytology (92% of cases). Controls were apparently healthy children enrolled contemporaneous to the cases from the nearest neighbor house to the case house and were age,- sex frequency-matched to the cases. Anti-SE36 IgG antibodies were measured using enzyme-linked absorbent immunoassays (ELISAs). SE36 titers were estimated by extrapolating ELISA optical density readings to a standard fitting curve. Anti SE36 titers were log-transformed for analysis. Odds ratios (ORs) and two-sided 95% confidence intervals (95% CIs) were estimated using unconditional logistic regression. The mean log endpoint dilution titers were 0.63 logs lower in cases than in controls (8.26 [SD 1.68] vs. 8.89 [SD 1.75], Student's t-test, p = 0.019). Lower titers were observed in cases than controls aged 0-4 years (p = 0.05) and in those aged 5-14 years (p = 0.06). Low and medium tertiles of anti SE36 IgG antibodies were associated with increased OR for BL ([OR 1.67, 95% CI 1.21-2.31] and [OR 1.33, 95% CI 0.96-1.86], respectively, p(trend) = 0.002) in analyses adjusting for age, sex, calendar period and test plate. Our findings suggest that compared to similarly aged children enrolled from the same community, children with BL in Ghana have lower antibodies to SE36 antigen. PMID- 21630257 TI - High throughput sequencing reveals diversity of Human Papillomaviruses in cutaneous lesions. AB - There are at least 120 completely characterized human papillomavirus (HPV) types and putative new types are continuously found. Both squamous cell carcinoma of the skin (SCC) and other skin lesions commonly contain multiple cutaneous HPV types. The objective of this study was to achieve an improved resolution of the diversity of HPV types in lesions such as SCCs, actinic keratoses (AKs) and keratoacanthomas (KAs). Fresh frozen biopsies from 37 SCC lesions, 36 AK lesions and 92 KA lesions and swab samples from the top of the lesion from 86 SCCs and 92 AKs were amplified using the general HPV primers FAP and mixed to three pools followed by high throughput sequencing. We obtained 2196 reads with homology to HPV. In the pool of SCC/AK biopsies 48 different HPV types were found. Eighty three types were found in the pool of SCC/AK swab samples and 64 types in the KA biopsies, respectively. For 9 novel putative HPV types most of the amplimer sequence was obtained, whereas for an additional 35 novel putative HPV types only partial amplimer sequences were obtained. Most of the novel putative types belonged to the genus Gamma. In conclusion, high throughput sequencing was an effective means to identify both known and previously unknown HPV types in putatively HPV-associated lesions and has revealed an extended diversity of HPV types. PMID- 21630258 TI - Mammographic density and risk of breast cancer by adiposity: an analysis of four case-control studies. AB - The association of mammographic breast density with breast cancer risk may vary by adiposity. To examine effect modification by body mass index (BMI), the authors standardized mammographic density data from four case-control studies (1994-2002) conducted in California, Hawaii and Minnesota and Gifu, Japan. The 1,699 cases and 2,422 controls included 45% Caucasians, 40% Asians and 9% African Americans. Using ethnic-specific BMI cut points, 34% were classified as overweight and 19% as obese. A single reader assessed density from mammographic images using a computer-assisted method. Logistic regression was used to estimate odds ratios (OR) and 95% confidence intervals (95% CI) while adjusting for potential confounders. Modest heterogeneity in the relation between percent density and breast cancer risk across studies was observed (p(heterogeneity) = 0.08). Cases had a greater age-adjusted mean percent density than controls: 31.7% versus 28.5%, respectively (p <0.001). Relative to <20 percent density, the ORs for >35 were similar across BMI groups whereas the OR for 20-35 was slightly higher in overweight (OR = 1.69, 95% CI: 1.28, 2.24) and obese (OR = 1.62, 95% CI: 1.12, 2.33) than in normal weight women (OR = 1.49, 95% CI: 1.11, 2.01). Furthermore, limited evidence of effect modification by BMI of the OR per 10% increase in percent density (p(interaction) = 0.06) was observed, including subgroup analyses by menopausal status and in analyses that excluded women at the extremes of the BMI scale. Our findings indicate little, if any, modification by BMI of the effects of breast density on breast cancer risk. PMID- 21630259 TI - Breast cancer prognosis is inherited independently of patient, tumor and treatment characteristics. AB - Population-based studies have shown a concordance of breast cancer survival among first-degree relatives (FDRs), suggesting a heritable component. Reasons for such heritability remain to be elucidated. We aimed to determine whether association of breast cancer survival among FDRs is linked to shared patient and tumor characteristics or type of treatment. At the population-based Geneva Breast Cancer Registry, we identified 162 FDR pairs diagnosed with breast cancer. We categorized FDRs into poor, medium and good familial survival risk groups according to breast cancer-specific survival of their proband (mother or sister). We compared patient, tumor and treatment characteristics between categories and calculated standardized mortality ratios (SMRs) and adjusted disease-specific mortality for each group. Breast cancer patients in the poor familial survival risk group were more likely to be diagnosed at later stages than those in the good familial survival risk group. Similarly, they had higher SMRs than those in the medium and good survival risk groups (18.7, 95% confidence interval [CI]: 9.4 33.5 vs. 16.5, 95% CI: 7.5-31.3 and 9.4, 95% CI: 3.4-20.4, respectively). After adjustment for patient and tumor characteristics and type of treatment, women in the poor familial survival risk group were almost five times more likely to die of breast cancer than those in the good familial survival risk group (adjusted hazard ratio 4.8, 95% CI: 1.4-16.4). Our study shows that breast cancer prognosis clusters within families and suggests that the hereditary component is independent of patient and tumor characteristics and type of treatment. PMID- 21630260 TI - JNK1 and JNK2 play redundant functions in Myc-induced B cell lymphoma formation. PMID- 21630262 TI - T cells raised against allogeneic HLA-A2/CD20 kill primary follicular lymphoma and acute lymphoblastic leukemia cells. AB - T cells mediating a graft-versus-leukemia/lymphoma effects without causing graft versus-host disease would greatly improve the safety and applicability of hematopoietic stem cell transplantation. We recently demonstrated that highly peptide- and HLA-specific T cells can readily be generated against allogeneic HLA A*02:01 in complex with a peptide from the B cell-restricted protein CD20. Here, we show that such CD20-specific T cells can easily be induced from naive precursors in cord blood, demonstrating that they do not represent cross-reactive memory cells. The cells displayed high avidity and mediated potent cytotoxic effects on cells from patients with the CD20(pos) B cell malignancies follicular lymphoma (FL) and acute lymphoblastic leukemia (ALL). However, the cytotoxicity was consistently lower for cells from two of the ALL patients. The ALL cells that were less efficiently killed did not display lower surface expression of CD20 or HLA-A*02:01, or mutations in the CD20 sequence. Peptide pulsing fully restored the levels of cytotoxicity, indicating that they are indeed susceptible to T cell mediated killing. Adoptive transfer of CD20-specific T cells to an HLA A*02:01(pos) patient requires an HLA-A*02:01(neg) , but otherwise HLA identical, donor. A search clarified that donors meeting these criteria can be readily identified even for patients with rare haplotypes. The results bear further promise for the clinical utility of CD20-specific T cells in B cell malignancies. PMID- 21630261 TI - The neurotensin receptor-1 promotes tumor development in a sporadic but not an inflammation-associated mouse model of colon cancer. AB - Neurotensin receptor-1 (NTR-1) is overexpressed in colon cancers and colon cancer cell lines. Signaling through this receptor stimulates proliferation of colonocyte-derived cell lines and promotes inflammation and mucosal healing in animal models of colitis. Given the causal role of this signaling pathway in mediating colitis and the importance of inflammation in cancer development, we tested the effects of NTR-1 in mouse models of inflammation-associated and sporadic colon cancer using NTR-1-deficient (Ntsr1(-) (/-)) and wild-type (Ntsr1(+/+)) mice. In mice treated with azoxymethane (AOM) to model sporadic cancer, NTR-1 had a significant effect on tumor development with Ntsr1(+/+) mice developing over twofold more tumors than Ntsr1(-) (/-) mice (p = 0.04). There was no effect of NTR-1 on the number of aberrant crypt foci or tumor size, suggesting that NT/NTR-1 signaling promotes the conversion of precancerous cells to adenomas. Interestingly, NTR-1 status did not affect tumor development in an inflammation-associated cancer model where mice were treated with AOM followed by two cycles of 5% dextran sulfate sodium (DSS). In addition, colonic molecular and histopathologic analyses were performed shortly after a single cycle of DSS. NTR 1 status did not affect colonic myeloperoxidase activity or histopathologic scores for damage and inflammation. However, Ntsr1(-) (/-) mice were more resistant to DSS-induced mortality (p = 0.01) and had over twofold higher colonic expression levels of Il6 and Cxcl2 (p < 0.04), cytokines known to promote tumor development. These results represent the first direct demonstration that targeted disruption of the Ntsr1 gene reduces susceptibility to colon tumorigenesis. PMID- 21630263 TI - Overexpression of NaV 1.6 channels is associated with the invasion capacity of human cervical cancer. AB - Functional activity of voltage-gated sodium channels (VGSC) has been associated to the invasion and metastasis behaviors of prostate, breast and some other types of cancer. We previously reported the functional expression of VGSC in primary cultures and biopsies derived from cervical cancer (CaC). Here, we investigate the relative expression levels of VGSC subunits and its possible role in CaC. Quantitative real-time PCR revealed that mRNA levels of Na(V) 1.6 alpha-subunit in CaC samples were ~40-fold higher than in noncancerous cervical (NCC) biopsies. A Na(V) 1.7 alpha-subunit variant also showed increased mRNA levels in CaC (~20 fold). All four Na(V) beta subunits were also detected in CaC samples, being Na(V) beta1 the most abundant. Proteins of Na(V) 1.6 and Na(V) 1.7 alpha-subunits were immunolocalized in both NCC and CaC biopsies and in CaC primary cultures as well; however, although in NCC sections proteins were mainly relegated to the plasma membrane, in CaC biopsies and primary cultures the respective signal was stronger and widely distributed in both cytoplasm and plasma membrane. Functional activity of Na(V) 1.6 channels in the plasma membrane of CaC cells was confirmed by whole-cell patch-clamp experiments using Cn2, a Na(V) 1.6-specific toxin, which blocked ~30% of the total sodium current. Blocking of sodium channels VGSC with tetrodotoxin and Cn2 did not affect proliferation neither migration, but reduced by ~20% the invasiveness of CaC primary culture cells in vitro assays. We conclude that Na(V) 1.6 is upregulated in CaC and could serve as a novel molecular marker for the metastatic behavior of this carcinoma. PMID- 21630264 TI - The age-specific prevalence of human papillomavirus and risk of cytologic abnormalities in rural Nigeria: implications for screen-and-treat strategies. AB - Cervical screening for carcinogenic human papillomavirus (HPV) infection is being considered for low-income countries. Effectiveness requires targeted screening in older women in whom prevalent infections are more likely to be persistent and predictive of precancer. Some studies in West Africa have found unusually high HPV prevalences across all adult ages, which may reduce the positive predictive value (PPV) of HPV-based screening, if positivity in older women does not sufficiently predict elevated risk. We conducted a population-based study in rural Nigeria to identify HPV prevalence and associated cervical abnormalities. Using stratified random sampling, we enrolled women age 15+. Nonvirgins had a cervical exam including liquid-based cytology and PCR HPV DNA testing from residual cytology specimens. Two-thirds of invited women participated, and 14.7% had detectable carcinogenic HPV, a proportion that did not decline with age (p trend = 0.36) and showed slight peaks in the 15-29 and 60-69 age groups. Among women of the age typically considered for screen-and-treat programs (30-49 years), 12.8% were HPV positive, and the PPV for high-grade or worse cytology was 16.4%. Comparatively, women age < 30 were more likely to be HPV positive (18.9%, p = 0.03) with a lower PPV (4.2% p = 0.05). Among women age 50+ (typically excluded from screening in resource-poor settings because inexpensive treatment is not available), HPV positivity was 14.2% with a PPV of 13.9%. In Irun and similar settings where HPV does not decline with age, HPV-based screen-and-treat programs might be feasible for mid-adult women because prevalence is sufficiently low and positivity predicts elevated risk of more easily treated precancer. PMID- 21630265 TI - The interplay between lipid profiles, glucose, BMI and risk of kidney cancer in the Swedish AMORIS study. AB - With exception of cholesterol and total fat intake, associations between lipid biomarkers and kidney cancer have not often been researched. We aimed to assess possible links between lipid profiles and kidney cancer risk in a large prospective cohort study, while also taking into account glucose levels and BMI. A cohort based on 542,924 persons with baseline information on glucose, triglycerides (TGs), total cholesterol (TC) and creatinine was selected from the Swedish Apolipoprotein Mortality Risk study. A subgroup of 85,621 also had baseline measurements of HDL, LDL, apolipoprotein A-I and apoB. Multivariate Cox proportional hazard models were used to analyze associations between quartiles and dichotomized values of these lipid components and kidney cancer risk. During a mean follow-up of 13 years, 958 persons developed kidney cancer. TGs were the only lipid component for which a statistically significant association was found with kidney cancer risk when using both quartiles and a clinical cutoff (hazard ratio: 1.25 (95% CI: 0.99-1.60), 1.29 (1.01-1.66) and 1.66 (1.30-2.13) for the 2nd, 3rd and 4th quartile, compared to the 1st, with p-value for trend: <0.001). The association remained after exclusion of the 95% percentile of TG. Quartiles of glucose were also positively associated with kidney cancer risk, whereas quartiles of TC were negatively associated with kidney cancer risk. This detailed analysis of lipid components only showed a consistent relation between TG levels and kidney cancer risk. Further mechanistic studies are required to assess links between lipid abnormalities and kidney cancer. PMID- 21630266 TI - Bistable switch in migration stimulating factor expression: regulation by the concerted signalling of transforming growth factor-beta1 and the extracellular matrix. AB - Migration stimulating factor (MSF) is an oncofetal motogenic/angiogenic cytokine constitutively expressed by epithelial and stromal cells in fetal and neoplastic tissues. Fibroblasts derived from healthy adult skin do not express MSF but can be induced to do so by treatment with transforming growth factor-beta1 (TGF beta1). As the bioactivities of both MSF and TGF-beta1 are modulated by the extracellular matrix, we investigated whether the induction of MSF expression by TGF-beta1 is also matrix dependent. We now report that adult fibroblasts are induced to express MSF by a transient treatment with TGF-beta1 (as short as 2 hr) but only when the cells are adherent to a "wound" matrix, such as denatured type I collagen, fibrin or plastic tissue culture dishes. Unexpectedly, this induction of MSF expression persists unabated for the entire subsequent lifespan of the treated cells in the absence of further TGF-beta1 and irrespective of the substratum. Such "activated" MSF expression may, however, be persistently switched off again by a second transient exposure to TGF-beta1 but this time only when the cells are adherent to a "healthy" matrix of native type I collagen. Significantly, the constitutive expression of MSF by fetal and cancer patient fibroblasts could also be persistently switched off by this means. We conclude that TGF-beta1 may both switch on and switch off MSF expression in a manner critically determined by the nature of the matrix substratum and suggest that this may be a possible mechanism underlying the observed dual functionality of TGF-beta1 as both a tumour suppressor and tumour promoter. PMID- 21630267 TI - Integrin targeted oncolytic adenoviruses Ad5-D24-RGD and Ad5-RGD-D24-GMCSF for treatment of patients with advanced chemotherapy refractory solid tumors. AB - The safety of oncolytic viruses for treatment of cancer has been shown in clinical trials while antitumor efficacy has often remained modest. As expression of the coxsackie-adenovirus receptor may be variable in advanced tumors, we developed Ad5-D24-RGD, a p16/Rb pathway selective oncolytic adenovirus featuring RGD-4C modification of the fiber. This allows viral entry through alpha-v-beta integrins frequently highly expressed in advanced tumors. Advanced tumors are often immunosuppressive which results in lack of tumor eradication despite abnormal epitopes being present. Granulocyte-macrophage colony stimulating factor (GMCSF) is a potent activator of immune system with established antitumor properties. To stimulate antitumor immunity and break tumor associated immunotolerance, we constructed Ad5-RGD-D24-GMCSF, featuring GMCSF controlled by the adenoviral E3 promoter. Preliminary safety of Ad5-D24-RGD and Ad5-RGD-D24 GMCSF for treatment of human cancer was established. Treatments with Ad5-D24-RGD (N = 9) and Ad5-RGD-D24-GMCSF (N = 7) were well tolerated. Typical side effects were grade 1-2 fatigue, fever and injection site pain. 77% (10/13) of evaluable patients showed virus in circulation for at least 2 weeks. In 3 out of 6 evaluable patients, disease previously progressing stabilized after a single treatment with Ad5-RGD-D24-GMCSF. In addition, 2/3 patients had stabilization or reduction in tumor marker levels. All patients treated with Ad5-D24-RGD showed disease progression in radiological analysis, although 3/6 had temporary reduction or stabilization of marker levels. Induction of tumor and adenovirus specific immunity was demonstrated with ELISPOT in Ad5-RGD-D24-GMCSF treated patients. RGD modified oncolytic adenoviruses with or without GMCSF seem safe for further clinical development. PMID- 21630268 TI - Role of exosomes released by chronic myelogenous leukemia cells in angiogenesis. AB - Our study is designed to assess if exosomes released from chronic myelogenous leukemia (CML) cells may modulate angiogenesis. We have isolated and characterized the exosomes generated from LAMA84 CML cells and demonstrated that addition of exosomes to human vascular endothelial cells (HUVEC) induces an increase of both ICAM-1 and VCAM-1 cell adhesion molecules and interleukin-8 expression. The stimulation of cell-cell adhesion molecules was paralleled by a dose-dependent increase of adhesion of CML cells to a HUVEC monolayer. We further showed that the treatment with exosomes from CML cells caused an increase in endothelial cell motility accompanied by a loss of VE-cadherin and beta-catenin from the endothelial cell surface. Functional characterization of exosomes isolated from CML patients confirmed the data obtained with exosomes derived from CML cell line. CML exosomes caused reorganization into tubes of HUVEC cells cultured on Matrigel. When added to Matrigel plugs in vivo, exosomes induced ingrowth of murine endothelial cells and vascularization of the Matrigel plugs. Our results suggest for the first time that exosomes released from CML cells directly affect endothelial cells modulating the process of neovascularization. PMID- 21630269 TI - MicroRNA profiling for the identification of cancers with unknown primary tissue of-origin. AB - Cancer of unknown primary (CUP) represents a common and important clinical problem. There is evidence that most CUPs are metastases of carcinomas whose primary site cannot be recognized. Driven by the hypothesis that the knowledge of primary cancer could improve patient's prognosis, we investigated microRNA expression profiling as a tool for identifying the tissue of origin of metastases. We assessed microRNA expression from 101 formalin-fixed, paraffin embedded (FFPE) samples from primary cancers and metastasis samples by using a microarray platform. Forty samples representing ten different cancer types were used for defining a cancer-type-specific microRNA signature, which was used for predicting primary sites of metastatic cancers. A 47-miRNA signature was identified and used to estimate tissue-of-origin probabilities for each sample. Overall, accuracy reached 100% for primary cancers and 78% for metastases in our cohort of samples. When the signature was applied to an independent published dataset of 170 samples, accuracy remained high: correct prediction was found within the first two options in 86% of the metastasis cases (first prediction was correct in 68% of cases). This signature was also applied to predict 16 CUPs. In this group, first predictions exhibited probabilities higher than 90% in most of the cases. These results establish that FFPE samples can be used to reveal the tissue of origin of metastatic cancers by using microRNA expression profiling and suggest that the approach, if applied, could provide strong indications for CUPs, whose correct diagnosis is presently undefined. PMID- 21630270 TI - Clopidogrel protects from cell apoptosis and oxidative damage in a mouse model of renal ischaemia-reperfusion injury. AB - Renal ischaemia-reperfusion injury (IRI) is consecutive to tissue oxidative damage and cell apoptosis that lead to acute renal failure (ARF) in renal allografts. The aim of this study was to investigate the beneficial effects of a pretreatment by clopidogrel on renal IRI in mice. IRI was induced by bilateral renal ischaemia for 45 min followed by reperfusion. Sixty-two healthy male BALB/c mice were randomly assigned to one of the following groups: PBS + ischaemia reperfusion (IR); clopidogrel + IR; PBS + sham IR; clopidogrel + sham IR. Clopidogrel (25 mg/kg) or PBS was administered per os to the animals via a gastric cannula 24 h before operation. All mice were given a single dose of clopidogrel or PBS. Renal function histological damage, renal cell apoptosis, renal antioxidant activities, and CD41 expression were determined 24 h after reperfusion. The survival rates were evaluated over 7 days. Animals pretreated with clopidogrel had lower plasma levels of blood urea nitrogen (BUN) and creatinine, lower histopathological scores, and improved survival rates following IR. Renal cell apoptosis induced by IR was decreased in kidneys of mice pretreated by clopidogrel, with an increase in Bcl-2 and Bcl-xL expression and a decrease in caspase-3, caspase-8, and Bax expression. Renal reduced glutathione, superoxide dismutase, and catalase activities were unmodified by the pretreatment with clopidogrel. However, clopidogrel resulted in an increased total antioxidant capacity of the kidney. Furthermore, pretreatment by clopidogrel decreased the number of CD41-positive cells. Thus, clopidogrel exerts protective effects on renal IRI in mice by abrogating renal cell apoptosis as a consequence of improved renal antioxidant capacity and could be tried as a novel therapeutic tool in renal IRI. PMID- 21630271 TI - Reduced Paneth cell antimicrobial protein levels correlate with activation of the unfolded protein response in the gut of obese individuals. AB - The intestinal microbiota is increasingly acknowledged to play a crucial role in the development of obesity. A shift in intestinal microbiota composition favouring the presence of Firmicutes over Bacteroidetes has been observed in obese subjects. A similar shift has been reported in mice with deficiency of active Paneth cell alpha-defensins. We aimed at investigating changes in Paneth cell antimicrobial levels in the gut of obese subjects. Next, we studied activation of the unfolded protein response (UPR) as a possible mechanism involved in altered Paneth cell function. Paneth cell numbers were counted in jejunal sections of 15 severely obese (BMI > 35) and 15 normal weight subjects. Expression of Paneth cell antimicrobials human alpha-defensin 5 (HD5) and lysozyme were investigated using immunohistochemistry, qPCR, and western blot. Activation of the UPR was assessed with western blot. Severely obese subjects showed decreased protein levels of both HD5 and lysozyme, while Paneth cell numbers were unchanged. Lysozyme protein levels correlated inversely with BMI. Increased expression of HD5 (DEFA5) and lysozyme (LYZ) transcripts in the intestine of obese subjects prompted us to investigate a possible translational block caused by UPR activation. Binding protein (BiP) and activating transcription factor 4 (ATF4) levels were increased, confirming activation of the UPR in the gut of obese subjects. Furthermore, levels of both proteins correlated with BMI. Involvement of the UPR in the lowered antimicrobial protein levels in obese subjects was strongly suggested by a negative correlation between BiP levels and lysozyme levels. Additionally, indications of ER stress were apparent in Paneth cells of obese subjects. Our findings provide the first evidence for altered Paneth cell function in obesity, which may have important implications for the obesity-associated shift in microbiota composition. In addition, we show activation of the UPR in the intestine of obese subjects, which may underlie the observed Paneth cell compromise. PMID- 21630272 TI - Nephrin expression in adult rodent central nervous system and its interaction with glutamate receptors. AB - Nephrin is an immunoglobulin-like adhesion molecule first discovered as a major component of the podocyte slit diaphragm, where its integrity is essential to the function of the glomerular filtration barrier. Outside the kidney, nephrin has been shown in other restricted locations, most notably in the central nervous system (CNS) of embryonic and newborn rodents. With the aim of better characterizing nephrin expression and its role in the CNS of adult rodents, we studied its expression pattern and possible binding partners in CNS tissues and cultured neuronal cells and compared these data to those obtained in control renal tissues and podocyte cell cultures. Our results show that, besides a number of locations already found in embryos and newborns, endogenous nephrin in adult rodent CNS extends to the pons and corpus callosum and is expressed by granule cells and Purkinje cells of the cerebellum, with a characteristic alternating expression pattern. In primary neuronal cells we find nephrin expression close to synaptic proteins and demonstrate that nephrin co-immunoprecipitates with Fyn kinase, glutamate receptors and the scaffolding molecule PSD95, an assembly that is reminiscent of those made by synaptic adhesion molecules. This role seems to be confirmed by our findings of impaired maturation and reduced glutamate exocytosis occurring in Neuro2A cells upon nephrin silencing. Of note, we disclose that the very same nephrin interactions occur in renal glomeruli and cultured podocytes, supporting our hypothesis that podocytes organize and use similar molecular intercellular signalling modules to those used by neuronal cells. PMID- 21630273 TI - Expression of hypoxia-associated proteins in sporadic medullary thyroid cancer is associated with desmoplastic stroma reaction and lymph node metastasis and may indicate somatic mutations in the VHL gene. AB - Medullary thyroid carcinomas (MTCs) are mostly aggressive but slowly growing malignant tumours that metastasize early to loco-regional lymph nodes. Desmoplastic stroma reaction is a strong risk factor associated with lymph node metastases. We evaluated immunohistochemically the expression of two hypoxia associated proteins, carbonic anhydrase IX (CAIX) and hypoxia-induced factor 1alpha (HIF1alpha), and ki-67, intercellular matrix adhesion molecule E-cadherin and the stroma remodelling marker tenascin C in a series of 100 sporadic MTCs and corresponding lymph node metastases, if present. Moderate to strong expression of CAIX was seen in 53 cases, and of HIF1alpha in 51 cases, showing a strong correlation (p < 0.001; Spearman's coefficient of correlation, 0.59). Expression correlated with the degree of desmoplasia (pCAIX = 0.001 and pHIF1alpha = 0.001), with tenascin C expression (pCAIX = 0.001,pHIF1alpha = 0.038), with the ki-67 proliferation index (pCAIX = 0.001, pHIF1alpha = 0.001) and with the presence of lymph node metastases (pCAIX < 0.001 and pHIF1alpha = 0.007). The absence of membranous E-cadherin staining was significantly associated with the grade of desmoplasia, tenascin expression and lymph node metastases(p <= 0.05) but not with ki67 proliferation index or expression of hypoxia-associated factors. Expression of hypoxia-associated proteins was in most cases identical between primary tumours and lymph node metastases.Two cases showed strong uniform expression of CAIX and HIF1alpha in the primary tumour as well as in the lymph node metastases, and sequencing revealed mutations in the coding regions of the Von-Hippel-Lindau gene (VHL ).Our findings suggest that despite of the fact that MTCs have only slowly growth, tumour hypoxia plays an important role in the development of loco-regional metastases. Since traditional cytotoxic chemotherapy has only little effect on MTCs, targeting hypoxia-associated and -regulated proteins might be of benefit for patients. PMID- 21630275 TI - Solute carrier transporters as targets for drug delivery and pharmacological intervention for chemotherapy. AB - Many solute carrier transporters that interact with anticancer agents and contribute to their pharmacokinetics have been shown to be differentially upregulated in cancer cells as a result of adaptive response to altered nutritional requirements. This review focuses on pathophysiological function of membrane transporters responsible for the influx of physiological substances including oligopeptides, amino acids, and organic cations and anions, and summarizes the recent knowledge regarding mechanisms in their gene expressions. Broad substrate specificity of enhanced oligopeptide H(+) /peptide cotransporter 1 activity in cancer cells is useful for tumor tissue-specific delivery of chemotherapeutic agents and positron emission tomography diagnostic probes. Amino acid transporters such as LAT1 and ASCT2 are upregulated in human cancer cells and are thought to stimulate tumor growth by regulating mammalian target of rapamycin through nutrient pathway. Especially, LAT1 could be a molecular target to deprive cancer cells of amino acids in combination with aminopeptidase inhibitors. As organic anion transporting polypeptides carry estrone-3-sulfate that is intracellularly hydrolyzed to estrone, their overexpression may provide a pharmacological merit to treat hormone-responsive breast tumors. Therefore, it is important to understand the pathophysiological significance and gene expression in cancer to develop new rationales for drug delivery and pharmacological interventions for chemotherapy. PMID- 21630274 TI - Aberrant succination of proteins in fumarate hydratase-deficient mice and HLRCC patients is a robust biomarker of mutation status. AB - Germline mutations in the FH gene encoding the Krebs cycle enzyme fumarate hydratase predispose to hereditary leiomyomatosis and renal cell cancer (HLRCC) syndrome. FH-deficient cells and tissues accumulate high levels of fumarate, which may act as an oncometabolite and contribute to tumourigenesis. A recently proposed role for fumarate in the covalent modification of cysteine residues to S (2-succinyl) cysteine (2SC) (termed protein succination) prompted us to assess 2SC levels in our existing models of HLRCC. Herein, using a previously characterized antibody against 2SC, we show that genetic ablation of FH causes high levels of protein succination. We next hypothesized that immunohistochemistry for 2SC would serve as a metabolic biomarker for the in situ detection of FH-deficient tissues. Robust detection of 2SC was observed in Fh1 (murine FH)-deficient renal cysts and in a retrospective series of HLRCC tumours (n = 16) with established FH mutations. Importantly, 2SC was undetectable in normal tissues (n = 200) and tumour types not associated with HLRCC (n = 1342). In a prospective evaluation of cases referred for genetic testing for HLRCC, the presence of 2SC-modified proteins (2SCP) correctly predicted genetic alterations in FH in every case. In two series of unselected type II papillary renal cancer (PRCC), prospectively analysed by 2SCP staining followed by genetic analysis, the biomarker accurately identified previously unsuspected FH mutations (2/33 and 1/36). The investigation of whether metabolites in other tumour types produce protein modification signature(s) that can be assayed using similar strategies will be of interest in future studies of cancer. PMID- 21630276 TI - Solubility prediction of drugs in mixed solvents using partial solubility parameters. AB - Solubility of drugs in binary and ternary solvent mixtures composed of water and pharmaceutical cosolvents at different temperatures were predicted using the Jouyban-Acree model and a combination of partial solubility parameters as interaction descriptors in the solution. The generally trained version of the model produced the overall mean percentage deviation values for the back calculated solubility of drugs in binary solvents of 34.3% and the predicted solubilities in ternary solvent mixtures of 38.0%. In addition, the applicability of the trained model for predicting the solvent composition providing the maximum solubility of a drug was investigated. The results of collected solubility data of drugs in various mixed solvents and the newly measured solubility data of five drugs in ethanol + propylene glycol + water mixtures at 25 degrees C showed that the model provided acceptable predictions and could be used in the pharmaceutical industry. PMID- 21630277 TI - Inhibition of crystallization and growth of celecoxib nanoparticles formed from volatile microemulsions. AB - The inhibitory effect of ammonium glycyrrhizinate (AG) on crystallization of celecoxib (CXB) nanoparticles in aqueous medium was studied. CXB nanoparticles in powder form were prepared by rapid evaporation of all solvents from a volatile oil-in-water microemulsion. A powder containing 13 wt % CXB was obtained by immediate conversion of microemulsion droplets into nanoparticles by spray drying. CXB was amorphous in this powder, which could be easily dispersed 1 wt % in water as nanoparticles. However, these particles crystallized rapidly upon dispersion, and a significant particle growth was observed. The natural surfactant, AG, which is US Food and Drug Administration approved for oral administration, inhibited crystallization of CXB, enabling a stable dispersion of nanoparticles with average size of 14 nm. Molecular dynamics simulations revealed rapid attachment of glycyrrhizinate to the growing CXB crystal and suggested that crystallization inhibition is due to interactions between the hydrophobic part of glycyrrhizinate and the phenyl moieties of CXB. PMID- 21630278 TI - Lead PK commentary: predicting human pharmacokinetics. PMID- 21630279 TI - Professor Akira Tsuji: scientist, educator, and leader. PMID- 21630280 TI - Solubility advantage of amorphous pharmaceuticals, part 3: Is maximum solubility advantage experimentally attainable and sustainable? AB - A method is described for screening compounds that inhibit crystallization in solution to enable more accurate measurement of amorphous drug solubility. Three polymers [polyvinylpyrrolidone, hydroxypropyl methylcellulose, and hydroxypropyl methylcellulose acetate succinate (HPMCAS)] were screened for their ability to inhibit the crystallization of neat amorphous drugs during measurement of solubility of the amorphous form in water. Among the polymers evaluated, HPMCAS was found to be most promising. The use of HPMCAS provided an "apparent solubility" of amorphous drugs that was closer to the theoretically calculated values. With danazol, agreement was essentially quantitative, and for griseofulvin and iopanoic acid, agreement was within a factor of two; these maximum concentrations were sustained for a period of 40-90 min. Dynamic light scattering of filtered samples (0.22 u) revealed the presence of colloidal drug polymer assemblies in solution (100-150 nm). The supernatant resulting from this centrifugation gradually decreased in concentration, but remained supersaturated with respect to crystalline drug for several hours. Thus, HPMCAS has been shown to be a useful additive in dissolution media to allow a more accurate determination of the solubility of fast crystallizing neat amorphous drugs, at least for the drugs studied, and it should also serve to retard crystallization in vivo and therefore, facilitate improved bioavailability. PMID- 21630281 TI - Pharmacokinetics of SN2310, an injectable emulsion that incorporates a new derivative of SN-38 in patients with advanced solid tumors. AB - SN2310 is an injectable emulsion composed of vitamin E, a succinate derivative, as well as 7-ethyl-10-hydroxycamptothecin (SN-38), the active metabolite of irinotecan. Single intravenous doses of 15, 20, 25, and 30 mg/m(2) of SN2310 emulsion were administered in a total of 26 patients with advanced solid malignancies. Serial blood samples were collected and concentrations of SN2310, SN-38, and SN-38 glucuronide were assayed. Mean systemic clearance of SN2310 ranged between 1.91 and 2.02 L/h/m(2) . Peak concentrations of SN-38 were observed at the end of infusion, suggesting a fast metabolic conversion of SN2310 to its active form, SN-38. Mean t1/2 values of SN-38 across the 20-30 mg/m(2) dose levels (131-199 h) were 33-55-fold longer than those observed for SN2310. The systemic exposure of SN-38 increased in a proportional manner over the dose range studied. SN2310 emulsion displayed an improved safety profile as compared with irinotecan. The most significant safety risk was neutropenia. Considering the rapid formation of SN-38, the proportional increase in exposure levels, and its longer elimination half-life, less frequent dosing of SN2310 emulsion may be considered for the treatment of patients with advanced solid malignancies. PMID- 21630282 TI - Mechanism for distribution of acotiamide, a novel gastroprokinetic agent for the treatment of functional dyspepsia, in rat stomach. AB - The novel gastroprokinetic agent acotiamide improves gastric motility by inhibiting acetylcholinesterase activity in stomach; however, the mechanism of distribution of acotiamide from blood to stomach has not been clarified. Here, the tissue distribution of acotiamide was investigated in rats. The tissue-to plasma concentration ratio (K(p,app,in vivo)) for stomach decreased from 4.1 to 2.4 mL/g of tissue at steady state with increasing plasma concentrations, whereas the K(p,app,in vivo) for skeletal muscle was much lower and constant, regardless of the concentration of acotiamide in plasma. In vitro binding to stomach tissue protein exhibited a linear profile, with a predicted K(p,app,in vitro) of 2.2 from free fractions under linear conditions. Therefore, protein binding to stomach tissue might only play a limited role in the stomach distribution of acotiamide. The influx permeability (f (u,b) * PS(inf,app)) in the stomach exhibited dose-dependent saturation at the lowest range of examined blood unbound concentrations of acotiamide, whereas that in skeletal muscle exhibited only minimal dose dependence. In addition, the unbound concentration ratio of stomach to plasma (2.8) at steady state was markedly higher than unity. Taken together, these results suggest that carrier-mediated concentrative uptake processes play an important role in the distribution of acotiamide to the stomach but not skeletal muscle. PMID- 21630283 TI - Nest membership determines the levels of aggression and cooperation between females of a supposedly communal digger wasp. AB - Females of communal and eusocial Hymenoptera differ greatly in the treatment of conspecifics: communal species lack the nest membership-bias treatment via nestmate (NM) recognition, which is typical of eusocial Hymenoptera. Therefore, an analysis of social interactions can suggest whether a given bee or wasp is communal or eusocial. For this reason, we observed females of the digger wasp Cerceris rubida in experimental dyadic encounters. Because this species had previously been labeled "communal," we expected to find little evidence of a NM bias with respect to cooperation or aggression when paired with NMs and non nestmates (NNM). Contrary to expectation, females were highly cooperative toward NMs and highly aggressive toward NNMs. Variation in the intensity of aggression toward NNMs partially stems from the degree to which the paired females differed in size. From these results and quantitative comparisons with studies on solitary, communal, and eusocial bee species, it appears that C. rubida has a more complex social organization than previously believed and, in particular, that females of this species exhibit NM recognition, a trait associated with eusociality. PMID- 21630284 TI - Fraction absorbed (Fabs ): different connotations and confusion for the literature? PMID- 21630285 TI - The coexistence of multiple cardiovascular diseases is an independent predictor of the 30-day mortality of hospitalized patients with congestive heart failure: a study in Beijing. AB - BACKGROUND: Congestive heart failure (CHF) has become a major clinical and public health challenge with the aging of the population in China. However, the effect of the coexistence of multiple cardiovascular diseases on the prognosis of hospitalized patients with CHF remains unclear. A comparative analysis was performed to explore the etiology and comorbidities of CHF and in-hospital mortality in patients with CHF. HYPOTHESIS: The coexistence of multiple cardiovascular diseases is an independent predictor of the 30-day mortality of hospitalized patients with CHF. METHODS: All 6949 patients (4344 males, 2605 females) in the database with a principal ICD-9-CM discharge diagnosis of CHF were identified and divided into 5 groups according to the number of comorbidities. Cox proportional hazards regression was used to examine the association between the number of comorbidities and in-hospital mortality in patients with CHF. RESULTS: A single comorbidity was predominant in younger patients (18-39 years, P<0.001), whereas multiple comorbidities were predominant in the elderly (>=80 years, P<0.001). The most common single etiology was valvular heart disease, and the most common triple etiology was hypertension and diabetes mellitus complicated by coronary artery disease. Cox regression analysis showed a higher hospital mortality rate associated with an increased number of comorbidities (hazard ratio [HR] from 1.22 [95% CI: 0.89-1.68] to HR 3.49 [95% CI: 2.55-4.78], P<0.001). CONCLUSIONS: This study demonstrates the coexistence of multiple cardiovascular diseases is an independent predictor of the 30-day mortality of hospitalized patients with CHF. PMID- 21630286 TI - Modeling for accurate dimensional scanning electron microscope metrology: then and now. AB - A review of the evolution of modeling for accurate dimensional scanning electron microscopy is presented with an emphasis on developments in the Monte Carlo technique for modeling the generation of the electrons used for imaging and measurement. The progress of modeling for accurate metrology is discussed through a schematic technology timeline. In addition, a discussion of a future vision for accurate SEM dimensional metrology and the requirements to achieve it are presented. PMID- 21630287 TI - Surface modification of vitreoretinal surgical instruments with layer-by-layer films. AB - Commercially available vitreoretinal surgical forceps were modified with layer-by layer (LbL) films designed to render them the ability to specifically adhere human the inner limiting membrane. Surgical forceps with two different geometries were etched, polished, and silanized before deposition of the films composed of poly (allylamine hydrochloride), poly (styrene sulfonate), and cationic gold nanoparticles. Stability and integrity of the LbL films was scrutinized by exposing the modified forceps to commercial disinfectant Cidex-OPA and then placing the instruments in a physiological-like HEPES buffer (pH 7.4, 5 mM, 154 mM NaCl). Surface topography analysis with scanning electron microscopy revealed that the geometry of the surgical instrument may affect the integrity of the film. Analysis of the HEPES buffer with inductively coupled plasma mass spectrometry demonstrated that gold nanoparticles did not leach from the LbL film after 60 min. PMID- 21630288 TI - Long-term structural changes of plasmid DNA studied by atomic force microscopy. AB - Long-term stability of plasmid DNA (pDNA) conformations is critical in many research areas, especially those concerning future gene therapy. Despite its importance, the time-evolution of pDNA structures has rarely been studied at a molecular resolution. Here, the time-evolution of pDNA solutions spanning four years was observed with atomic force microscopy (AFM). The AFM data show that the pDNA molecules changed over time from isolated supercoiled structures, to aggregated supercoiled structures, to thin, branched network structures, and finally to wider, branched network structures. Additional topographical analysis of the AFM data suggests that the actions of residual proteins could be the main mechanism for the structural changes in our laboratory-prepared pDNA. PMID- 21630290 TI - Genotyping of Saccharomyces cerevisiae strains by interdelta sequence typing using automated microfluidics. AB - Amplification of genomic sequences flanked by delta elements of retrotransposons TY1 and TY2 is a reliable method for characterization of Saccharomyces cerevisiae strains. The aim of this study is to evaluate the usefulness of microfluidic electrophoresis (Caliper LabChip) to assess the factors that affect interlaboratory reproducibility of interdelta sequence typing for S. cerevisiae strain delimitation. We carried out experiments in two laboratories, using varying combinations of Taq DNA polymerases and thermal cyclers. The reproducibility of the technique is evaluated using non-parametric statistical tests and we show that the source of Taq DNA polymerase and technical differences between laboratories have the highest impact on reproducibility, whereas thermal cyclers have little impact. We also show that the comparative analysis of interdelta patterns is more reliable when fragment sizes are compared than when absolute and relative DNA concentrations of each band are considered. Interdelta analysis based on a smaller fraction of bands with intermediate sizes between 100 and 1000 bp yields the highest reproducibility. PMID- 21630291 TI - PROCOS: computational analysis of protein-protein complexes. AB - One of the main challenges in protein-protein docking is a meaningful evaluation of the many putative solutions. Here we present a program (PROCOS) that calculates a probability-like measure to be native for a given complex. In contrast to scores often used for analyzing complex structures, the calculated probabilities offer the advantage of providing a fixed range of expected values. This will allow, in principle, the comparison of models corresponding to different targets that were solved with the same algorithm. Judgments are based on distributions of properties derived from a large database of native and false complexes. For complex analysis PROCOS uses these property distributions of native and false complexes together with a support vector machine (SVM). PROCOS was compared to the established scoring schemes of ZRANK and DFIRE. Employing a set of experimentally solved native complexes, high probability values above 50% were obtained for 90% of these structures. Next, the performance of PROCOS was tested on the 40 binary targets of the Dockground decoy set, on 14 targets of the RosettaDock decoy set and on 9 targets that participated in the CAPRI scoring evaluation. Again the advantage of using a probability-based scoring system becomes apparent and a reasonable number of near native complexes was found within the top ranked complexes. In conclusion, a novel fully automated method is presented that allows the reliable evaluation of protein-protein complexes. PMID- 21630293 TI - Preoperative serum levels of early prostate cancer antigen (EPCA) predict prostate cancer progression in patients undergoing radical prostatectomy. AB - BACKGROUND: Early prostate cancer antigen (EPCA) has been shown a prostate cancer (PCa)-associated nuclear matrix protein, however, its serum status and prognostic power in patients with PCa are unknown. The goals of this study are to measure preoperative serum EPCA levels in a cohort of PCa patients who were treated with radical prostatectomy (RP), and to investigate whether serum EPCA levels would independently predict cancer prognosis after the surgery. METHODS: The study group consisted of 109 consecutive patients with clinically localized PCa who were candidates for RP. Serum EPCA levels were measured by ELISA prior to the surgery, and were correlated with pathologic parameters and clinical outcomes postoperatively. RESULTS: A total of 106 patients underwent RP. Preoperative mean serum EPCA level in RP patients (15.84 +/- 3.63 ng/ml) was significantly higher than that in healthy subjects (4.62 +/- 1.15 ng/ml) (P < 0.001), but serum EPCA levels in the both groups were statistically lower than the levels in patients with PCa metastatic to regional lymph nodes (27.83 +/- 6.22 ng/ml) and metastatic to bone (28.50 +/- 6.67 ng/ml) (all P's < 0.001). In patients who progressed during follow-up, preoperative serum mean EPCA levels were higher in those with aggressive disease progression (27.64 +/- 5.48 ng/ml) compared with nonaggressive disease progression (18.15 +/- 4.63 ng/ml; P < 0.001). In pre- and postoperative multivariate analyses, preoperative serum EPCA level was an independent predictor for disease progression (Hazards Ratio = 5.016, P < 0.001 and Hazards Ratio = 4.305, P < 0.001, respectively). CONCLUSIONS: Preoperative serum EPCA level is significantly elevated in localized PCa patients with metastatic disease and strongly predicts cancer progression postoperatively. PMID- 21630292 TI - Serum prosaposin levels are increased in patients with advanced prostate cancer. AB - BACKGROUND: We previously cloned prosaposin (PSAP) from metastatic castrate resistant prostate cancer (mCRPCa) cells and demonstrated its genomic amplification and/or overexpression in metastatic PCa cell lines, xenografts, and lymph node metastases. The clinicohistopathological significance of serum PSAP levels and its tissue expression and association with predictive or prognostic variable in primary or advanced PCa are not known. METHODS: We examined PSAP expression by immunohistochemical staining during early embryogenic development of the prostate and within a large tissue microarray which included 266 benign and malignant prostate tissues. In addition, serum PSAP levels in the age adjusted normal male population and in 154 normal individuals and patients with primary or mCRPCa were measured by an ELISA assay. RESULTS: Univariate and multivariate analyses revealed a significant and inverse association between PSAP expression and clinical stages II and III tumors, dominant Gleason patterns 3 and 4, and seminal vesicle invasion. In the normal male population, the lowest serum PSAP level was detected before puberty, peaked at the most reproductive age group (20- to 39-year old), and then, decreased to a range between the two groups for men above 40-year old. Regardless of age and when compared with normal individuals, serum PSAP levels significantly decreased in primary organ-confined PCa, but increased in those with mCRPCa. CONCLUSION: Our results show that PSAP has the potential to differentiate between primary and advanced PCa. Additional large-scale studies are needed to define the usefulness of tissue expression or serum PSAP levels as a diagnostic or prognostic marker or as a therapeutic target in PCa. PMID- 21630294 TI - Use of tumor dynamics to clarify the observed variability among biochemical recurrence nomograms for prostate cancer. AB - BACKGROUND: Nomograms for biochemical recurrence (BCR) of prostate cancer (PC) after radical prostatectomy can yield very different prognoses for individual patients. Since the nomograms are optimized on different cohorts, the variations may be due to differences in patient risk-factor distributions. In addition, the nomograms assign different relative scores to the same PC risk factors and rarely stratify for tumor growth rate. METHODS: We compared BCR-free probabilities from the GPSM model with a cell kinetics (CK) model that uses the individual's tumor state and growth rate. We first created a cohort of 143 patients that reproduced the GPSM patient distribution in Gleason score, Prostate specific antigen (PSA), Seminal vesicle involvement and Margin status since they form the GPSM score. We then performed 143 CK calculations to determine BCR-free probabilities for comparison with the GPSM results for all scores and with four other prominent nomograms for a high-risk patient. RESULTS: The BCR-free probabilities from the CK model agree within 10% with those from the GPSM study for all scores once the CK model parameters are stratified in terms of the GPSM risk factors and the PSA doubling time (PSADT). However, the probabilities from widely used nomograms vary significantly. CONCLUSIONS: The CK model reproduces the observed GPSM BCR-free probabilities with a broad stratification of model parameters for PC risk factors and can thus be used to describe PC progression for individual patients. The analysis suggests that nomograms should stratify for PSADT to be predictive. PMID- 21630295 TI - Androgens repress expression of the F-box protein Skp2 via p107 dependent and independent mechanisms in LNCaP prostate cancer cells. AB - BACKGROUND: Androgens control homeostasis of the normal prostate and growth of prostate cancer (PCa) through the androgen receptor (AR) by regulating gene networks involving in cell proliferation, differentiation, and survival. We demonstrated previously that expression of Skp2, a key protein regulating cell entry into the S phase, is inhibited by androgens in an AR-dependent manner (Oncogene, 2004; 23(12): 2161-2176). However, the underlying mechanism of this regulation is unknown. METHODS: Using the LNCaP PCa cell line as a working model, the effect of androgens on the expression of Skp2 was examined by Western and Northern blot analyses. Cell cycle was measured by fluorescence-activated cell sorting (FACS). Gene transfection was performed by electroporation to manipulate the expression levels of proteins studied. RESULTS: At physiological levels androgens markedly repressed Skp2 expression but slightly induced Skp2 expression at subphysiological levels. Androgens modestly decreased the stability of the Skp2 protein. Androgenic repression of Skp2 expression was completely abolished by E1A-mediated inactivation of pocket proteins including RB, p130, and p107. Moreover, ectopic expression of p107 inhibited Skp2 expression, and silencing of p107 partially blocked androgenic repression of Skp2. CONCLUSIONS: Our data indicate that androgens repress Skp2 expression via p107-dependent and independent pathways in PCa cells. These regulatory mechanisms may be targeted for the development of new therapeutics of androgen-refractory PCa. PMID- 21630296 TI - The need for pathological confirmation of the diagnosis of peritoneal malignant mesothelioma. PMID- 21630297 TI - Peer coaching and mentoring: a new model of educational intervention for safe patient handling in health care. AB - BACKGROUND: To reduce the risk of patient handling-related musculoskeletal injury, overhead ceiling lifts have been installed in health care facilities. To increase ceiling lift usage for a variety of patient handling tasks, a peer coaching and mentoring program was implemented among the direct care staff in the long-term care subsector in British Columbia, Canada. They received a 4-day training program on body mechanics, ergonomics, patient-handling techniques, ceiling lift usage, in addition to coaching skills. METHODS: A questionnaire was administered among staff before and after the intervention to evaluate the program's effectiveness. RESULTS: There were 403 and 200 respondents to the pre intervention and post-intervention questionnaires. In general, staff perceived the peer-coaching program to be effective. The number of staff who reported to be using ceiling lifts "often and always" went higher from 64.5% to 80.5% (<0.001) after coaching program implementation. Furthermore, staff reported that they were using the ceiling lifts for more types of tasks post-intervention. Staff reported that the peer coaching program has increased their safety awareness at work and confidence in using the ceiling lifts. CONCLUSIONS: The findings suggest that this educational model can increase the uptake of mechanical interventions for occupational health and safety initiatives. It appears that the training led to a greater awareness of the availability of or increased perceptions of the number of ceiling lifts, presumably through coaches advocating their use. PMID- 21630298 TI - Elevated serum liver enzymes and fatty liver changes associated with long driving among taxi drivers. AB - BACKGROUND: Previous studies suggested increased morbidities and mortalities of liver diseases in drivers. METHODS: To examine whether driving (monthly driving distance; tenure) is associated with elevated alanine aminotransferase (ALT), aspartate aminotransferase (AST), or chronic fatty liver (FL) changes, we performed a cross-sectional, secondary analysis of the Taxi Drivers' Health Study (n = 1,355), adjusting for clinical, demographic, and lifestyle factors. RESULTS: Prevalence of elevated ALT, elevated AST, and fatty liver changes were 22.0%, 5.1%, and 9.3%, respectively. Driving distance had a positive association with elevated ALT with a prevalence ratio of 1.35 (95% CI: 0.98, 1.89) comparing the highest versus lowest driving quartile. This association differed by alcohol use, with a corresponding prevalence ratio of 2.08 (95% CI: 1.30, 3.33) among "past/current" drinkers but no association among "never" drinkers. Similar patterns were found for AST, but estimates were less stable. We found a curvilinear response pattern for fatty liver changes; prevalence first increased with years as a taxi driver and then receded in the highest ranges of driving tenure, regardless of the alcohol history. CONCLUSIONS: Our results provide evidence that long driving is associated with both short-term and chronic liver insults, although alcohol use appears to modify this putative effect. PMID- 21630299 TI - Blood oxidative stress in Taiwan workers exposed to carbon disulfide. AB - BACKGROUND: Overexposure to carbon disulfide (CS(2) ) has been associated with an increase in coronary heart disease, but the mechanisms mediating this effect remain unclear. We aimed to examine the relationship between CS(2) exposure and oxidative stress markers, in order to clarify the oxidative mechanisms involved in CS(2) -induced atherosclerosis. METHODS: A total of 89 workers from a viscose rayon plant were recruited for this study, and 111 workers not exposed to CS(2) served as controls. Cholesterol, triglyceride, malondialdehyde (MDA), superoxide dismutase (SOD), catalase, GSH peroxidase, as well as total antioxidants were analyzed. RESULTS: The workers exposed to CS(2) had significantly higher MDA levels and lower SOD levels than the controls. The average MDA levels were 776 +/ 268.2 (240-1,220) in the high exposure (>=10 ppm; n = 38), 751.6 +/- 274 (170 1,320) in the low exposure (<10 ppm; n = 51), and 550.4 +/- 199 (115-1,050) mM in the control group (n = 111). The average SOD levels were 36.5 +/- 38.8 (0-223.5), 39.3 +/- 38.8 (0-160), and 58.8 +/- 60.8 (5.25, 400) U/ml in the high exposure-, low exposure-, and control group, respectively. MDA level increased significantly at a cumulative CS(2) exposure of over 60 ppm-years. Dyslipoproteinemia was borderline significantly associated with CS(2) exposure and MDA level. CONCLUSIONS: These results indicate that CS(2) exposure can induce oxidative stress as well as reduce the levels of antioxidative enzymes, and that a cumulative exposure level of 60 ppm-years may be a threshold value for the oxidative and the antioxidant response. Am. J. Ind. Med. 54:637-645, 2011. (c) 2011 Wiley-Liss, Inc. PMID- 21630300 TI - Incidence of work and non-work related disability claims in Brazil. AB - BACKGROUND: Sickness benefit claims are an important economic burden to society. This study aims to determine the incidence of sickness benefit claims in Brazil in 2008, exploring the role of economic activity. METHODS: Population-based study on sickness claims lasting longer than 15 days of sickness absence granted to private sector employees. Data on gender, age, economic activity, diagnosis, and work-relatedness were collected. RESULTS: The annual incidence of sickness benefits was 421.8/10,000 jobs, 435.4 for males and 452.0 for females. There were 3.5 times more non-work-related than work-related claims. The main diagnoses were injuries, musculoskeletal disorders, and mental disorders. Rates increased with age up to 59 years. Economic activity 37-Sewage had the highest incidence of non work-related and work-related claims. CONCLUSION: The incidence of sickness benefits is higher among female and older workers. Economic activities show great variability of sickness benefit rates, work-relatedness, diagnostic categories, and gender. PMID- 21630301 TI - Genotyping single nucleotide polymorphisms (SNPs) across species in Old World Monkeys. AB - The development of DNA markers is becoming increasingly useful in the field of primatology for studies on paternity, population history, and biomedical research. In this study, we determine the efficacy of using cross-species amplification to identify single nucleotide polymorphisms (SNPs) in closely related species. The DNA of 93 individuals representing seven Old World Monkey species was analyzed to identify SNPs using cross-species amplification and genotyping. The loci genotyped were 653 SNPs identified and validated in rhesus macaques. Of the 653 loci analyzed, 27% were estimated to be polymorphic in the samples studied. SNPs identified at the same locus among different species (coincident SNPs) were found in six of the seven species studied with longtail macaques exhibiting the highest number of coincident SNPs (84). The distribution of coincident SNPs among species is not biased based on proximity to genes in the samples studied. In addition, the frequency of coincident SNPs is not consistent with expectations based on their phylogenetic relationships. This study demonstrates that cross-species amplification and genotyping using the Illumina Golden Gate Array is a useful method to identify a large number of SNPs in closely related species, although issues with ascertainment bias may limit the type of studies where this method can be applied. PMID- 21630302 TI - Fetal hemoglobin in sickle cell anemia: Saudi patients from the Southwestern province have similar HBB haplotypes but higher HbF levels than African Americans. PMID- 21630303 TI - Blindness in a patient with chronic lymphocytic leukemia. PMID- 21630304 TI - Biomarkers for early detection of sickle nephropathy. AB - Renal complications affect nearly 30-50% of adults with sickle cell anemia (SCA), causing significant morbidity and mortality. Standard renal function tests like serum creatinine and glomerular filtration rate become abnormal in this disease only when renal damage has become extensive and largely irreversible. Moreover, not all patients develop sickle nephropathy (SN). Therefore, noninvasive biomarkers that predict early onset of SN are necessary. We performed a cross sectional analysis for nephropathy in 116 patients with sickle cell disease, analyzing urinary kidney injury molecule-1 (KIM-1), liver-type fatty acid binding protein (L-FABP), N-acetyl-b-D-glucosaminidase (NAG), neutrophil gelatinase associated lipocalin (NGAL) and transforming growth factor-beta1 (TGF-beta), together with conventional renal biomarkers (urine albumin and osmolality, and serum creatinine and cystatin C estimated GFR) during routine clinic visits when patients were at steady-state/baseline. We observed a distinct biomarker pattern: KIM-1 and NAG emerged as biomarkers with a strong association with albuminuria. Surprisingly, and in contrast to other acute/chronic renal disorders, NGAL, L FABP, and TGF-beta levels did not show any relationship with albuminuria in patients with SCA. Our study identifies potential biomarkers for SN, and suggests longitudinal validation of these biomarkers for early detection of SN, so that therapeutic interventions can be applied before renal damage becomes irreversible. PMID- 21630305 TI - A phase I safety study of enzastaurin plus bortezomib in the treatment of relapsed or refractory multiple myeloma. AB - The purpose of this study was to assess the safety and identify the recommended doses of enzastaurin and bortezomib in combination for future Phase II studies in patients with relapsed or refractory multiple myeloma. Three dose levels (DLs) of oral enzastaurin and intravenous bortezomib were used according to a conventional "3 + 3" design. A loading dose of enzastaurin (250 mg twice/day [BID]) on Day 1 was followed by enzastaurin 125 mg BID for 1 week, after which bortezomib was added (Cycle 1, 28 days, 1.0 mg/m(2) : Days 8, 11, 15, and 18; seven subsequent 21-day cycles, 1.3 mg/m(2) : Days 1, 4, 8, and 11). Twenty-three patients received treatment; all patients received prior systemic therapy. Most patients received >=3 regimens; 17 patients were bortezomib-refractory. A median of four treatment cycles (range 1-24) was completed. No dose-limiting toxicities were observed; thus, DL 3 was the recommended Phase II dose. The most common drug related Grade 3/4 toxicities were thrombocytopenia (n = 6) and anemia (n = 2). No patients died on therapy. One patient (DL 1) achieved a very good partial response; three patients (DLs 2 and 3), a partial response; nine patients, stable disease; and four patients, progressive disease. The recommended Phase II doses in patients with relapsed or refractory multiple myeloma are as follows: enzastaurin loading dose of 375 mg three times/day on Day 1 followed by 250 mg BID, with bortezomib 1.3 mg/m(2) on Days 1, 4, 8, and 11 of a 21-day cycle. The combination was well-tolerated and demonstrated some antimyeloma activity. PMID- 21630306 TI - Positive HIV ELISA test, autoimmune hemolytic anemia, and generalized lymphadenopathy: a unifying diagnosis. PMID- 21630307 TI - Nilotinib treatment-associated peripheral artery disease and sudden death: yet another reason to stick to imatinib as front-line therapy for chronic myelogenous leukemia. PMID- 21630308 TI - Lenalidomide, cyclophosphamide and dexamethasone (CRd) for newly diagnosed multiple myeloma: results from a phase 2 trial. AB - The combination of lenalidomide and low-dose dexamethasone is an effective treatment for multiple myeloma (MM). Addition of alkylating agents to lenalidomide or thalidomide results in increased response rates and deeper responses. We designed this trial to study the combination of cyclophosphamide, lenalidomide, and dexamethasone (CRd) as initial therapy for MM. Fifty-three patients with previously untreated symptomatic MM was enrolled. Patients received 4-week treatment cycles consisting of lenalidomide (25 mg daily for 3 weeks), dexamethasone (40 mg weekly), and cyclophosphamide (300 mg/m(2) weekly for 3 weeks). A partial response or better was seen in 85% of patients including 47% with a very good partial response or better. The toxicities were manageable with over 80% of planned doses delivered; six patients went off study for toxicity. The median progression free survival (PFS) for the entire group was 28 months (95% CI: 22.7-32.6) and the overall survival (OS) at 2 years was 87% (95% CI: 78 96). Importantly, 14 patients with high-risk MM had similar PFS and OS as the standard-risk patients (n = 39). CRd is an effective and well-tolerated regimen for upfront therapy of MM with high response rates and excellent 2-year OS, and is suitable for long-term therapy. PMID- 21630309 TI - Effectiveness and safety of high-dose cyclophosphamide as salvage therapy for high-risk multiple myeloma and plasma cell leukemia refractory to new biological agents. PMID- 21630310 TI - Dysplastic basophils in the accelerated phase of chronic myelogenous leukemia. PMID- 21630311 TI - Thrombotic microangiopathy complicating pegylated interferon treatment of hepatitis C infection. PMID- 21630312 TI - Laboratory and echocardiography markers in sickle cell patients with leg ulcers. PMID- 21630313 TI - A prognostic model for short term adverse events in normotensive patients with pulmonary embolism. AB - Risk stratification of patients with PE has gained interest in terms of the identification of patients in whom treatment on an outpatient base can be considered. Previous studies are of limited value due to their focus on adverse clinical events within several months after diagnosis of PE. We developed a prognostic model, based on easily accessible, clinical, and laboratory parameters, to predict adverse events during the first 10 days after the diagnosis of acute PE. We have analyzed the data of 210 outpatients with confirmed PE. Collected data included medical history, pulse rate, blood pressure, NT-proBNP, and D-dimer concentrations. The primary outcome was the occurrence of adverse clinical events in a 10 day follow-up period. Our final prognostic model to predict short-term adverse events consists of NT-proBNP levels, D-dimer concentrations, pulse rate, and the occurrence of active malignancy; the total score ranges from 0 to 37 points. Patients with a low score (no active malignancy, pulse rate <90 bpm, NT-proBNP <500 pg/ml, and D-dimer <3,000 MUg/l FEU) have a 10-day adverse event risk <1.5%. This risk increases to over 30% in patients with a maximum score, based on high pulse rate, D-dimer concentrations, and NT-proBNP levels. Our prognostic model, once prospectively validated in an independent sample of patients, can be used in the early risk stratification of PE to estimate the risk of adverse events and to differentiate between candidates for in- or out- hospital treatment. PMID- 21630314 TI - IgM multiple myeloma: more on a rare and heterogeneous disease. PMID- 21630315 TI - Primary effusion lymphoma in a HIV-negative patient associated with hypogammaglobulinemia. PMID- 21630316 TI - Circulating and progenitor endothelial cells are abnormal in patients with different types of von Willebrand disease and correlate with markers of angiogenesis. AB - von Willebrand disease (VWD) is the most common inherited bleeding disorder and is caused by quantitative or qualitative defects of von Willebrand factor (VWF). VWF, synthesized by endothelium and megakaryocytes (MK), circulates in plasma and is present in subendothelium and platelets. Circulating endothelial cells (CEC) and progenitor endothelial cells (EPC) have been recently proposed as markers of peripheral and bone marrow-derived angiogenesis. To evaluate the association of CEC/EPC with known inherited defects of cellular and circulating VWF, we have measured the number of CEC/EPC together with cytokines involved in angiogenesis in different VWD types. A group of 74 patients was composed by the following VWD types: VWD1 (n = 22), VWD2A (n = 9), VWD2B (n = 19), VWD2M (n = 17), and VWD3 (n = 7). Healthy individuals (n = 20) were used as controls. CEC (CD146(+) , CD31(+) , and CD45(-) ) and EPC (CD34(+) , CD133(+) , and CD45(-) ) were evaluated by flow cytometry. Circulating serum levels of VEGF, E-selectin, P-selectin, EPO, and TPO were determined by ELISA. CEC, VEGF, E-selectin, and EPO were higher and EPC lower in VWD patients than in controls (P < 0.01). Among the five groups of VWD patients and controls, a significant difference was found for CEC (one-way ANOVA: P = 0.005), EPC (P = 0.001), E-Selectin (P < 0.0001), EPO (P = 0.021), and TPO (P = 0.004): the latter was high in VWD3 patients. In VWD1, we found an inverse relationship between CEC and VWF:Ag levels (P = 0.048; R(2) = 0.19). Based on these data, CEC are increased in VWD and are associated with the high levels of cytokines involved in angiogenesis (up-regulation). EPC are decreased, suggesting down-regulation of bone marrow-derived angiogenesis in VWD. PMID- 21630317 TI - Intra-arterial methylprednisolone for the management of steroid-refractory acute gastrointestinal and hepatic graft versus host disease. PMID- 21630318 TI - Gastric teeth of some thoracotreme crabs and their contribution to the brachyuran phylogeny. AB - The gastric teeth of three ocypodoid species were investigated using scanning electron microscopy, and the morphological results were discussed with respect to the known food preferences. The species were chosen in particular because of contrasting ideas about their relationships within the Thoracotremata. For the genera Heloecius, Dotilla, Mictyris, and "Uca" (s. str.), we find a specific correlation of the gastric teeth with the suspension feeding. The lateral gastric teeth of Uca have no prominent lateral teeth cusps, and most of their teeth surface consists of transverse comb-like lamellae. However, this possible food adaptation does not exclude the usability of specific teeth characters to distinguish species of suspension feeders. The closer relationship of the Dotillidae to grapsoid lines of gecarcinid or sesarmid crabs suggested by molecular data is not supported by the gastric teeth. For the genus Ucides, we found several characters that distinguish Ucides from the remaining ocypodoid genera Heloecius, Dotilla, Mictyris, and "Uca." In particular, the structures of the lateral and the dorsomedian teeth show some similarities to genera of the Gecarcinidae and Sesarmidae. Our results suggest that foregut characters can be used for phylogenetic analyses. PMID- 21630319 TI - Re-establishment of biting mouthparts in desert-living dung beetles (Scarabaeidae: Scarabaeinae) feeding on plant litter-Old structures reacquired or new ones evolved? AB - Evolution of mouthparts in adult dung beetles (Scarabaeidae: Scarabaeinae) for eating moist, fresh dung was linked with a loss of the ability to chew. However, the desert-living genus Pachysoma, probably evolved from a wet-dung feeding, Scarabaeus-like ancestor, has switched to a diet of dry fecal pellets (of rodents or small ruminants) and plant litter that requires re-establishment of chewing. Indeed, gut contents of a litter-feeding Pachysoma species indicate efficient food comminution. Based on scanning electron microscopy, cutting and grinding mouthpart structures in six Pachysoma species, of two lineages and with different food preferences, are described and compared with homologous structures in wet dung feeding Scarabaeus species. In Pachysoma, cutting and breaking of large food items is probably performed by a clypeal scraper, a prominent epipharyngeal tooth and large maxillary galeal hooks. Further comminution is achieved by a large, grinding area evolved on the mandibular molae. Interspecific differences and the probable function and evolution of these structures are discussed. Particularly, the unique tools for cutting/breaking are completely novel structures and not results of some reacquisition of normal biting mouthparts. PMID- 21630320 TI - Fine structure of the gnathosoma of archegozetes longisetosus [corrected] aoki (acari: oribatida, trhypochthoniidae). AB - Oribatida are one of the main groups of Acari comprising mostly important decomposers in soils. Most species are particle feeders, an exceptional mode of nutrition in Arachnida. Hence, their feeding organs, the gnathosoma, are of special functional interest. We studied nearly all components using scanning and transmission electron microscopies as well as reconstructions based on synchrotron X-ray microtomography from the model oribatid Archegozetes longisetosus. Besides cuticular structures, we describe the full set of muscles and confirm the presence of a trochanter remnant at the base of the chelicera. Setae on the prodorsum and the anterior and posterior infracapitular setae are mechanoreceptors innervated by two dendrites ending with tubular bodies. Dendrites of adoral setae, anterior setae of the chelicerae, and the supracoxal setae show neither obvious tubular bodies nor wall or terminal pores. Thus their function remains obscure. For the first time, a muscular proprioreceptor has been found in Arachnida. It likely monitors the actions of muscles moving the movable digit of the chelicera. Glandular structures within and associated with the gnathosoma are described. Dermal glands represented by secretory porose areas are found within the infracapitulum. More complex associated glands comprise the podocephalic glands and the infracapitular glands, the ducts of which were traced completely for the first time. The components described are mostly fundamental for the gnathosoma of Actinotrichida (Acariformes), one of the two lineages of Acari, to which Oribatida belong. The gnathosoma is generally considered the most relevant putative synapomorphy of Acari. Since the monophyly of Acari has become more and more questionable during the last decades, a thorough reinvestigation of this body part is necessary for a comprehensive understanding of acarine and even arachnid phylogeny and evolution. This article provides a starting point of such a re-evaluation of the gnathosoma. PMID- 21630321 TI - Finite element modeling of shell shape in the freshwater turtle Pseudemys concinna reveals a trade-off between mechanical strength and hydrodynamic efficiency. AB - Aquatic species can experience different selective pressures on morphology in different flow regimes. Species inhabiting lotic regimes often adapt to these conditions by evolving low-drag (i.e., streamlined) morphologies that reduce the likelihood of dislodgment or displacement. However, hydrodynamic factors are not the only selective pressures influencing organismal morphology and shapes well suited to flow conditions may compromise performance in other roles. We investigated the possibility of morphological trade-offs in the turtle Pseudemys concinna. Individuals living in lotic environments have flatter, more streamlined shells than those living in lentic environments; however, this flatter shape may also make the shells less capable of resisting predator-induced loads. We tested the idea that "lotic" shell shapes are weaker than "lentic" shell shapes, concomitantly examining effects of sex. Geometric morphometric data were used to transform an existing finite element shell model into a series of models corresponding to the shapes of individual turtles. Models were assigned identical material properties and loaded under identical conditions, and the stresses produced by a series of eight loads were extracted to describe the strength of the shells. "Lotic" shell shapes produced significantly higher stresses than "lentic" shell shapes, indicating that the former is weaker than the latter. Females had significantly stronger shell shapes than males, although these differences were less consistent than differences between flow regimes. We conclude that, despite the potential for many-to-one mapping of shell shape onto strength, P. concinna experiences a trade-off in shell shape between hydrodynamic and mechanical performance. This trade-off may be evident in many other turtle species or any other aquatic species that also depend on a shell for defense. However, evolution of body size may provide an avenue of escape from this trade off in some cases, as changes in size can drastically affect mechanical performance while having little effect on hydrodynamic performance. PMID- 21630322 TI - Functional morphology of the forelimb of living and extinct tree-kangaroos (Marsupialia: Macropodidae). AB - Tree-kangaroos are a unique group of arboreal marsupials that evolved from terrestrial ancestors. The recent discovery of well-preserved specimens of extinct tree-kangaroo species (genus Bohra) within Pleistocene cave deposits of south-central Australia provides a unique opportunity to examine adaptive evolution of tree-kangaroos. Here, we provide the first detailed description of the functional anatomy of the forelimb, a central component of the locomotor complex, in the extant Dendrolagus lumholtzi, and compare its structure and function with representatives of other extant marsupial families. Several features were interpreted as adaptations for coping with a discontinuous, uneven and three-dimensional arboreal substrate through enhanced muscular strength and dexterity for propulsion, grasping, and gripping with the forelimbs. The forelimb musculoskeletal anatomy of Dendrolagus differed from terrestrial kangaroos in the following principal ways: a stronger emphasis on the development of muscles groups responsible for adduction, grasping, and gripping; the enlargement of muscles that retract the humerus; and modified shape of the scapula and bony articulations of the forelimb bones to allow improved mobility. Many of these attributes are convergent with other arboreal marsupials. Tree-kangaroos, however, still retain the characteristic bauplan of their terrestrial ancestors, particularly with regard to skeletal morphology, and the muscular anatomy of the forelimb highlights a basic conservatism within the group. In many instances, the skeletal remains of Bohra have similar features to Dendrolagus that suggest adaptations to an arboreal habit. Despite the irony of their retrieval from deposits of the Nullarbor "Treeless" Plain, forelimb morphology clearly shows that the species of Bohra were well adapted to an arboreal habitat. PMID- 21630323 TI - Activity of the ovarian germinal epithelium in the freshwater catfish, Pimelodus maculatus (Teleostei: Ostariophysi: Siluriformes): germline cysts, follicle formation and oocyte development. AB - Distinct types of oogonia are found in the germinal epithelium that borders the ovarian lamellae of Pimelodus maculatus: A-undifferentiated, A-differentiated and B-oogonia. This is similar to the situation observed for spermatogonia in the vertebrate testis. The single A-undifferentiated oogonia divide by mitosis giving rise to A-groups of single differentiated oogonia, each enclosed by epithelial cells that are prefollicle cells. Subsequently, the single A-differentiated oogonia proliferate to generate B-oogonia that are interconnected by cytoplasmic bridges, hence, forming germline cysts. The prefollicle cells associated with them also divide. Within the germline cysts, B-oogonia enter meiosis becoming oocytes. Meiotic prophase and early folliculogenesis occur within the germline cysts. During folliculogenesis, prefollicle cells grow between the oocytes, encompassing and individualizing each of them. The intercellular bridges disappear, and the germline cysts are broken down. Next, a basement membrane begins to form around the nascent follicle, separating an oocyte and its associated prefollicle cells from the cell nest. Folliculogenesis is completed when the oocyte and the now follicle cells are totally encompassed by a basement membrane. Cells derived from the ovarian stroma encompass the newly-formed ovarian follicle, and become the theca, thereby completing the formation of the follicle complex. Follicle complexes remain attached to the germinal epithelium as they share a portion of basement membrane. This attachment site is where the oocyte is released during ovulation. The postovulatory follicle complex is continuous with the germinal epithelium as both are supported by a continuous basement membrane. The findings in P. maculatus reinforce the hypothesis that ovarian follicle formation represents a conserved process throughout vertebrate evolution. PMID- 21630324 TI - Morphological variation in head shape of pipefishes and seahorses in relation to snout length and developmental growth. AB - The feeding apparatus of Syngnathidae, with its elongate tubular snout and tiny, toothless jaws, is highly specialized for performing fast and powerful pivot feeding. In addition, the prolonged syngnathid parental care probably enables the juveniles to be provided with a feeding apparatus that resembles the one in adults, both in morphology and function. In this study, a landmark-based geometric morphometric analysis was carried out on the head of syngnathid representatives in order to (1) examine to what degree pipefish shape variation is different from that of seahorses; (2) determine whether the high level of specialization reduces the amount of intraspecific morphological variation found within the family; and (3) elucidate whether or not important shape changes occur in the seahorse head during postrelease ontogeny. We found that (1) there is a significant shape difference between head shape of pipefish and seahorse: the main differences concern snout length and height, position and orientation of the pectoral fin base, and height of the head and opercular bone. We hypothesize that this might be related to different prey capture kinematics (long snout with little head rotation versus short snout with large head rotation) and to different body postures (in line with the head versus vertical with a tilted head) in pipefishes and seahorses; (2) both pipefishes and seahorses showed an inverse relation between relative snout length and intraspecific variation and although pipefishes show a large diversity in relative snout elongation, they are more constrained in terms of head shape; and (3) the head of juvenile Hippocampus reidi specimens still undergoes gradual shape changes after being expelled from the brood pouch. Ontogenetic changes include lowering of the snout and head but also differences in orientation of the preopercular bone and lowering of the snout tip. PMID- 21630325 TI - Connexin43 interacts with betaarrestin: a pre-requisite for osteoblast survival induced by parathyroid hormone. AB - Parathyroid hormone (PTH) promotes osteoblast survival through a mechanism that depends on cAMP-mediated signaling downstream of the G protein-coupled receptor PTHR1. We present evidence herein that PTH-induced survival signaling is impaired in cells lacking connexin43 (Cx43). Thus, expression of functional Cx43 dominant negative proteins or Cx43 knock-down abolished the expression of cAMP-target genes and anti-apoptosis induced by PTH in osteoblastic cells. In contrast, cells lacking Cx43 were still responsive to the stable cAMP analog dibutyril-cAMP. PTH survival signaling was rescued by transfecting wild type Cx43 or a truncated dominant negative mutant of betaarrestin, a PTHR1-interacting molecule that limits cAMP signaling. On the other hand, Cx43 mutants lacking the cytoplasmic domain (Cx43(Delta245)) or unable to be phosphorylated at serine 368 (Cx43(S368A)), a residue crucial for Cx43 trafficking and function, failed to restore the anti-apoptotic effect of PTH in Cx43-deficient cells. In addition, overexpression of wild type betaarrestin abrogated PTH survival signaling in Cx43 expressing cells. Moreover, betaarrestin physically associated in vivo to wild type Cx43 and to a lesser extent to Cx43(S368A) ; and this association and the phosphorylation of Cx43 in serine 368 were reduced by PTH. Furthermore, induction of Cx43(S368) phosphorylation or overexpression of wild type Cx43, but not Cx43(Delta245) or Cx43(S368A) , reduced the interaction between betaarrestin and the PTHR1. These studies demonstrate that betaarrestin is a novel Cx43 interacting protein and suggest that, by sequestering betaarrestin, Cx43 facilitates cAMP signaling, thereby exerting a permissive role on osteoblast survival induced by PTH. PMID- 21630326 TI - Ghrelin induces cell migration through GHS-R, CaMKII, AMPK, and NF-kappaB signaling pathway in glioma cells. AB - Ghrelin is a newly discovered gastric peptide which stimulates food intake, energy balance, and growth hormone release. Recent reports have also shown that circulating ghrelin can efficiently reach the brain. However, the molecular mechanisms and pathophysiologic roles underlying ghrelin-induced glioma migration remain unclear. Glioma is the most common primary adult brain tumor with poor prognosis because of the spreading of tumor cell to the other regions of brain easily. In present study, we found that application of recombinant human ghrelin enhances the glioma cell migration in both rat C6 and human U251 cells. Ghrelin and its receptor GHS-R (growth hormone secretagogue receptor) are expressed in a wide variety of tissues and cell types, including various cancer cells. However, little is known about the expression of ghrelin or GHS-R in brain tumors. Here, we found that ghrelin increased GHS-R receptor up-regulation, and the enhancement of ghrelin-induced glioma cell motility markedly inhibited by a GHS-R antagonist. In addition, ghrelin-mediated migration was attenuated by treatment of CaMKII inhibitor, and AMPK inhibitors and pre-transfection with AMPK siRNA. Moreover, ghrelin stimulation also increased the phosphorylation of CaMKII and AMPK. Treatment with three different types of NF-kappaB inhibitors or pre-transfection with KM-IKKalpha, or KM-IKKbeta also reduced ghrelin-induced glioma cell migration. Moreover, treatment of ghrelin also induced IKKalpha/beta activation, IkappaBalpha phosphorylation, p65 phosphorylation at Ser(536), and increased NF kappaB-DNA binding activity and kappaB-transcriptional activity. These results indicate that ghrelin enhances migration of glioma cells is mainly regulated by the GHS-R, CaMKII, AMPK, and NF-kappaB pathway. PMID- 21630327 TI - Hyperglycemia regulates MDR-1, drug accumulation and ROS levels causing increased toxicity of carboplatin and 5-fluorouracil in MCF-7 cells. AB - There is constant increase in number of diabetic cases thereby giving it status of a serious epidemic. Diabetes increases the risk of occurrence of several cancers including breast cancer and may also have a serious impact on the outcome of cancer treatment. In the present study we investigated effect of hyperglycemia on cytotoxic efficacy of carboplatin and 5-fluorouracil in MCF-7 cells. MCF-7 cells were grown in 5.5 or 25 mM glucose chronically. We show that hyperglycemia favors proliferation of MCF-7 cells and increases expression of cell cycle regulatory proteins cyclin E and cdk-2. Hyperglycemia enhances cytotoxicity of carboplatin and 5-fluorouracil in MCF-7 cells by approximately 30% and decreases their IC50 by 1.5- and 1.3-folds, respectively. Hyperglycemia reduces expression of P-glycoprotein and promotes cell killing by increasing drug accumulation. Treatment with 40 uM verapamil, an inhibitor of P-gp activity specifically increases killing of MCF-7 cells cultured in 5.5 mM glucose. Further, this effect is synergized by elevated reactive oxygen species and treatment with N Acetylcysteine, an inhibitor of ROS, increases survival by 30 and 18% in carboplatin- and 5-fluorouracil-treated cells cultured in high glucose, respectively. Cytotoxicity of these drugs is associated with reduced activation of Akt and decreased transcriptional activation of NF-kappaB. In conclusion, hyperglycemia potentiates cytotoxicity of drugs by reducing P-gp expression and, increased ROS levels may partially or completely contribute to enhanced toxicity. PMID- 21630328 TI - Tissue engineering-based cartilage repair with mesenchymal stem cells in a porcine model. AB - This in vivo pilot study explored the use of mesenchymal stem cell (MSC) containing tissue engineering constructs in repair of osteochondral defects. Osteochondral defects were created in the medial condyles of both knees of 16 miniature pigs. One joint received a cell/collagen tissue engineering construct with or without pretreatment with transforming growth factor beta (TGF-beta) and the other joint from the same pig received no treatment or the gel scaffold only. Six months after surgery, in knees with no treatment, all defects showed contracted craters; in those treated with the gel scaffold alone, six showed a smooth gross surface, one a hypertrophic surface, and one a contracted crater; in those with undifferentiated MSCs, five defects had smooth, fully repaired surfaces or partially repaired surfaces, and one defect poor repair; in those with TGF-beta-induced differentiated MSCs, seven defects had smooth, fully repaired surfaces or partially repaired surfaces, and three defects showed poor repair. In Pineda score grading, the group with undifferentiated MSC, but not the group with TGF-beta-induced differentiated MSCs, had significantly lower subchondral, cell morphology, and total scores than the groups with no or gel only treatment. The compressive stiffness was larger in cartilage without surgical treatment than the treated area within each group. In conclusion, this preliminary pilot study suggests that using undifferentiated MSCs might be a better approach than using TGF-beta-induced differentiated MSCs for in vivo tissue engineered treatment of osteochondral defects. PMID- 21630329 TI - Characterization of a cartilage-like engineered biomass using a self-aggregating suspension culture model: molecular composition using FT-IRIS. AB - Maintenance of chondrocyte phenotype and robust expression and organization of macromolecular components with suitable cartilaginous properties is an ultimate goal in cartilage tissue engineering. We used a self-aggregating suspension culture (SASC) method to produce an engineered cartilage, "cartilage tissue analog" (CTA). With an objective of understanding the stability of phenotype of the CTA over long periods, we cultured chondrocytes up to 4 years and analyzed the matrix. Both early (eCTAs) (6 months) and aged (aCTAs) (4 years) showed type II collagen throughout with higher concentrations near the edge. Using Fourier transform-infrared imaging spectroscopy (FT-IRIS), proteoglycan/collagen ratio of eCTA was 2.8 times greater than native cartilage at 1 week, but the ratio was balanced to native level (p = 0.017) by 36 weeks. Surprisingly, aCTAs maintained the hyaline characteristics, but there was evidence of calcification within the tissue with a distinct range of intensities. Mineral/matrix ratio of those aCTA with "intensive" calcification was significantly higher (p = 0.017) than the "partial," but when compared to native bone the ratio of "intensive" aCTAs was 2.4 times lower. In this study we utilized the imaging approach of FT-IRIS and have shown that a biomaterial formed is compositionally closely related to natural cartilage for long periods in culture. We show that this culture platform can maintain a CTA for extended periods of time (4 years) and under those conditions signs of mineralization can be found. This method of cartilage tissue engineering is a promising method to generate cartilaginous biomaterial and may have potential to be utilized in both cartilage and boney repairs. PMID- 21630330 TI - In vivo strain analysis of the intact supraspinatus tendon by ultrasound speckles tracking imaging. AB - Limited data exist regarding why delaminated tears occur within the rotator cuff tendon, and no general agreement exists on how to handle this complicated tear. To analyze in vivo intratendinous strain of the supraspinatus tendon, the superficial, middle, and deep regions were marked with speckles using 2D speckle tracking echocardiography (2D STE) of 15 shoulders. The displacement and the strain of each speckle during isotonic and isometric shoulder motion were evaluated. Significantly different displacement and strains in the tendon were found between isometric and isotonic shoulder motions. In isometric motion, the average longitudinal displacement of the speckle at the superficial region (1.66 mm) was larger than at the deep region (0.61 mm), and the average peak strain at the superficial region (17.03%) was also higher than that at the deep region (3.42%). Conversely, in isotonic motion, the average longitudinal displacement of the speckle at the superficial region (0.70 mm) was less than that at the deep region (1.61 mm), and the average peak strain at the superficial region (4.73%) was also lower than that at the deep region (15.69%). A different strain was found between the superficial and deep regions within the intact live supraspinatus tendon. The strain and displacement patterns vary according to isometric versus isotonic shoulder motions. On the basis of our observations, we suggest that the delaminated tear of the rotator cuff tendon must be repaired separately layer by layer to resist the inhomogeneous strain after the repair. PMID- 21630331 TI - Long-term evaluation of a compliant cushion form acetabular bearing for hip joint replacement: a 20 million cycles wear simulation. AB - Soft bearing materials that aim to reproduce the tribological function of the natural joint are gaining popularity as an alternative concept to conventional hard bearing materials in the hip and knee. However, it has not been proven so far that an elastic cushion bearing can be sufficiently durable as a long term (~20 years) articulating joint prosthesis. The use of new bearing materials should be supported by accurate descriptions of the implant following usage and of the number, volume, and type of wear particles generated. We report on a long term 20 million cycle (Mc) wear study of a commercial hip replacement system composed of a compliant polycarbonate-urethane (PCU) acetabular liner coupled to a cobalt-chromium alloy femoral head. The PCU liner showed excellent wear characteristics in terms of its low and steady volumetric wear rate (5.8-7.7 mm(3)/Mc) and low particle generation rate (2-3 * 10(6) particles/Mc). The latter is 5-6 orders of magnitude lower than that of highly cross-linked polyethylene and 6-8 orders of magnitude lower than that of metal-on-metal bearings. Microscopic analysis of the implants after the simulation demonstrated a low damage level to the implants' articulating surfaces. Thus, the compliant PCU bearing may provide a substantial advantage over traditional bearing materials. PMID- 21630332 TI - A toenail flap based on the fibro-osseous hiatus branch for fingernail reconstruction. AB - This study included two parts: 1) cadaver dissection to elucidate the perfusion of toenail flaps by the fibro-osseous hiatus branch (FHB), and 2) clinical application of the toenail flap for reconstruction of a fingernail defect. Four second toes of two fresh Korean cadavers were dissected. The plantar digital artery (PDA) and terminal segment branch (TSB) were ligated, and red latex was injected distally into the ligated PDA. Perfusion of the dye into the toenail bed through the FHB was observed. From Oct 2004 to Sep 2009, eight toenail flaps based on the FHB pedicle with or without the distal phalanx and pulp were applied to seven patients for finger nail reconstruction. The toenail flap was marked at 5 mm distal to the nail fold and 5 mm lateral to the paronychium. The toenail complex based on the FHB was elevated and transferred to the finger. The nail and matrix were elevated with or without including the distal phalanx. The results of perfusion study showed that one side of the unilateral FHB was identified and traced proximal to the PDA, which was ligated. The distal toenail bed was perfused by the dye through the FHB. In clinical application, all the toenail flaps flourished and survived. We suggest that the toenail flap based on the FHB may be useful for fingernail reconstruction with minimal donor morbidity. PMID- 21630333 TI - Lower extremity free flap reconstruction outcomes using venous coupler. AB - BACKGROUND: Microvascular anastomotic coupling devices have been available to microsurgeons for over 20 years. Many studies have validated the efficacy of these devices for venous anastomosis. To date, there have been no large reports of their success in the anatomical region with the highest free flap failure rate, the lower extremity. METHODS: A retrospective review of 67 consecutive patients who underwent lower extremity microvascular reconstruction performed from August 2003 to September 2010 was performed. Patient charts were reviewed for age, sex, medical comorbidities, etiology of defect, location of defect, flap type, anastomotic technique, complications, flap survival, and limb salvage outcome. RESULTS: No patients returned to the operating room to have an arterial or venous anastomosis revised. Despite 100% vascular anastomosis patency rates in 67 consecutive lower extremity free flaps, flap survival rate was 95.5%. Total complication rate (13.4%) was due to two partial and one complete flap loss, three infections, two skin graft loses, and one hematoma. There were no intraoperative or perioperative complications involving the use of a microvascular anastomotic coupling device itself. Thirty-day and long term limb salvage rate was 97% and 92.5%, respectively. CONCLUSION: Microvascular anastomotic coupling devices create effective venous anastomoses in lower extremity microvascular reconstruction. Thus, it presents an important tool in the armamentarium for lower extremity microsurgical reconstruction. PMID- 21630334 TI - Mesh epineurial splinting for late median nerve repair in older patients: a preliminary report. AB - Eleven patients over 40 years old, with median nerve lesions at the wrist, were operated on an average of 5 months after their injury. In six patients, the median nerve was repaired using a polypropylene mesh applied to secure the nerve stumps in contact, thereby allowing for direct repair with microsutures. Six patients had their median nerve repaired with sural grafts. The average gap length was 2.8 cm for the mesh repair, whereas it was 3.7 cm for the graft repair group. Eighteen months after surgery, pressure thresholds were perceived in the index and thumb pulp by all six patients with a mesh repair but in only two of five patients with a graft repair. Five in the mesh repair group recovered function in the abductor pollicis brevis muscle, versus none in the graft group. These preliminary results suggest that, in older patients, the use of a mesh to splint the coaptation site followed by direct nerve repair yields better recovery than conventional sural grafts. PMID- 21630335 TI - Free tissue transfer in reconstruction following soft tissue sarcoma resection. AB - BACKGROUND: Radical surgical resection remains the single-most important treatment in the curative multimodal therapy of soft tissue sarcomas. Refinements in surgical techniques have resulted in the development of function preserving approaches increasingly avoiding limb amputation. PATIENTS AND METHODS: The records of all patients (n = 34) who underwent microsurgical soft tissue coverage subsequent to primary resection of soft tissue sarcoma of the upper or lower limb from 1999 to 2009 are reviewed regarding postoperative complications, time until start of adjuvant radiation and functional outcome (Toronto Extremity Salvage Score, TESS). RESULTS: Thirty-four patients (range: 21-86 years) received a total of 35 free flaps. Complete tumor resection was obtained in 33 patients, one patient required re-excision ultimately resulting in tumor-free margin status (R0 resection). Major complications were encountered in four cases including one patient with complete flap loss requiring an additional free flap and three patients with partial flap loss requiring split-thickness skin graft procedures. Minor complications were observed in three patients (9%). Extremity salvage could be achieved in 33 patients with adequate postoperative ambulation (TESS 84 +/- 18) and adequate use of the upper extremity (TESS 80 +/- 22). One patient underwent amputation. Mean time until start of adjuvant radiotherapy was 37 days (range 24-56 days). CONCLUSION: A synergetic center-based interdisciplinary approach is crucial in therapeutical management of soft tissue sarcomas with the aim of R0 resection status and limb preservation. Plastic surgery contributes by offering microsurgical reconstruction using free tissue transfer, thus broadening surgical possibilities. This increases the chance of both adequate oncosurgical resection and limb preservation. PMID- 21630336 TI - Intravenous heparin use in digital replantation and revascularization: The Quebec Provincial Replantation program experience. AB - BACKGROUND: No consensus exists among microsurgeons regarding the role of intravenous (IV) heparin in digital replantation/revascularization. The current experience of the Provincial Replantation Center in Quebec was reviewed over a 4 year period. METHODS: An initial retrospective review of all revascularized or reimplanted digits at our Replantation Center from April 2004 to April 2006 was conducted. Then, data of all patients treated at our center from January 08 to September 08 were prospectively collected. The two cohorts were compared with regards to demographics, injury characteristics, postoperative thromboprophylaxis medication as well as complication and success rates. Proportions were compared using chi(2) tests/Fisher's exact tests. Multivariate analysis was conducted with logistic regression. RESULTS: 175 digits were treated from April 2004 to April 2006, including 104 revascularizations and 71 amputations. IV heparin was used in 35.1% of the cases and was associated with a 3.59-fold (95% CI, 1.55-8.31) increase risk of developing a complication compared with cases where heparin was not used (P = 0.001). In 2008, 106 digits were treated. IV heparin was used in 14.6% of the cases and was not significantly associated with a higher complication rate compared with cases where heparin was not used (P = 0.612). Both cohorts' success rates were very similar (P = 0.557). The number of complications decreased from the first period (20.5%) to the second one (12.8%). CONCLUSION: Routine use of IV heparin following digital replantation and revascularization is not warranted. Surgical technique and type of injury remains the most important predictors for success in these complex procedures. PMID- 21630337 TI - The mesh repair: tension free alternative on dealing with nerve gaps-experimental results. AB - We evaluated the feasibility of external epineurial splinting as a way of alleviating tension caused by sutures in the reconstruction of peripheral nerve injuries, utilizing Wistar rat median nerve injury on 40 animals, in four experimental groups with 10 animals on each surgical setting. The nerve regeneration outcomes of four surgical procedures were compared: 1) primary end to-end sutures (EES); 2) alleviated tension sutures (ATS) with a removal of 7 mm nerve segment, namely external epineurial splinting, utilizing a polypropylene mesh as a protective scaffold; 3) sutures under tension with a 7 mm gap between nerve stumps; and 4) sham (C) (n = 10 animals). Regeneration of the median nerve postneurorrhaphy was followed by means of functional evaluations, including time to first day of finger flexion recovery, and grasp strength; quantification of atrophy of the flexor pronator muscle group; and mRNA expression of nerve growth factor and neurotrophin-3 by polymerase chain reaction-reverse transcriptase. Similar, significant median nerve regeneration was observed in the EES-treated and ATS-treated groups, relative to controls. The EES and ATS surgical procedures methods demonstrated important similar results considering functional and molecular biology analysis of the median nerve injury. PMID- 21630338 TI - Vascular complications and microvascular free flap salvage: the role of thrombolytic agents. AB - BACKGROUND: Vascular thrombosis with flap loss is the most dreaded complication of microvascular free tissue transfer. Thrombolytic agents such as tissue plasminogen activator have been used clinically for free flap salvage in cases of pedicle thrombosis. Yet, there is a paucity of data in the literature validating the benefit of their use. METHODS: A retrospective review of the breast reconstruction free flap database was performed at a single institution between the years of 1991-2010. The incidence of vascular complications (arterial and/or venous thrombosis) was examined to determine the role of adjuvant thrombolytic therapy in flap salvage. Pathologic examination was used to determine the incidence of fat necrosis after secondary revision procedures. RESULTS: Seventy four cases were identified during the study period. In 41 cases, revision of the anastamoses was performed alone without thrombolytics with 38 cases of successful flap salvage (92.7%). In 33 cases, anastamotic revision was performed with adjuvant thrombolytic therapy, and successful flap salvage occurred in 28 of these cases (84.8%). Thrombolysis did not appear to significantly affect flap salvage. Interestingly, only two of the salvaged flaps that had received thrombolysis developed fat necrosis, whereas 11 of the nonthrombolysed flaps developed some amount fat necrosis (7.1% vs. 28.9%, P < 0.05). CONCLUSIONS: The decreased incidence of fat necrosis may be attributable to dissolution of thrombi in the microvasculature with the administration of thrombolytics. Although the use of adjuvant thrombolytic therapy does not appear to impact the rate of flap salvage, their use may have secondary benefits on overall flap outcomes. PMID- 21630339 TI - Sternocleidomastoid region restoration with lateral hemisoleus muscle incorporatied in free fibular flap for reconstruction of radical neck dissection and hemimandibulectomy. AB - In the treatment of head and neck carcinoma, radical cervical lymphadenectomy leaves the affected side of the neck devoid of the sternocleidomastoid muscle, thus more vulnerable to the unwanted side effects of the adjuvant radiotherapy. It also causes asymmetry and cosmetically unpleasant appearance of the cervical region. In the reported case with widely ulcerated squamous-cell carcinoma over mandible, hemimandibulectomy and radical neck dissection was performed. Following the mandibular reconstruction, the lateral hemisoleus muscle of the harvested osteomyocutaneous fibula flap was utilized to restore the ipsilateral sternocleidomastoid region. This new application promises to be a useful method, which can aid in the restoration of the aesthetic contour of the neck and provide protection against unwanted effects of the adjuvant radiotherapy on the ipsilateral carotid artery. PMID- 21630340 TI - Changes in arterial blood flow of free flaps after the administration of sildenafil in swine. AB - We evaluated blood flow changes after experimental free tissue transfer and the potential hemodynamic effect of sildenafil on the free flap. Sixteen swine were used for free transfer of a latissimus dorsi myocutaneous flap to the chest that was anastomosed to the internal mammary vessels, and were randomized into controls and study group. The latter received a single dose of sildenafil, 6 hours following flap revascularization. Doppler ultrasonography revealed that arterial flow was mainly systolic postoperatively. Diastolic flow patterns were gradually restored after the first postoperative day. Pulsatility index (PI) significantly increased and flow volume decreased in all animals postoperatively. In the sildenafil group, PI significantly decreased and flow volume increased, while diastolic flow patterns were restored earlier on compared to controls, postoperatively. In conclusion, the administration of sildenafil after free tissue transfer increases flow volume and facilitates the restoration of diastolic blood flow patterns in the early critical postoperative period. PMID- 21630341 TI - Modeling of Look-Locker estimates of the magnetic resonance imaging estimate of longitudinal relaxation rate in tissue after contrast administration. AB - This paper models the behavior of the longitudinal relaxation rate of the protons of tissue water R(1) (R(1) = 1/T(1) ), measured in a Look-Locker experiment at 7 Tesla after administration of a paramagnetic contrast agent (CA). It solves the Bloch-McConnell equations for the longitudinal magnetization of the protons of water in a three-site two-exchange (3S2X) model with boundary conditions appropriate to repeated sampling of magnetization. The extent to which equilibrium intercompartmental water exchange kinetics affect monoexponential estimates of R(1) after administration of a CA in dynamic contrast enhanced experiment is described. The relation between R(1) and tissue CA concentration was calculated for CA restricted to the intravascular, or to the intravascular and extracellular compartments, by varying model parameters to mimic experimental data acquired in a rat model of cerebral tumor. The model described a nearly linear relationship between R(1) and tissue concentration of CA, but demonstrated that the apparent longitudinal relaxivity of CA depends upon tissue type. The practical consequence of this finding is that the extended Patlak plot linearizes the DeltaR(1) data in tissue with leaky microvessels, accurately determines the influx rate of the CA across these microvessels, but underestimates the volume of intravascular blood water. PMID- 21630342 TI - Design of a radiative surface coil array element at 7 T: the single-side adapted dipole antenna. AB - Ultra high field MR imaging (>=7 T) of deeply located targets in the body is facing some radiofrequency-field related challenges: interference patterns, reduced penetration depth, and higher Specific Absorbtion Ratio (SAR) levels. These can be alleviated by redesigning the elements of the transmit or transceive array. This is because at these high excitation field (B(1) ) frequencies, conventional array element designs may have become suboptimal. In this work, an alternative design approach is presented, regarding coil array elements as antennas. Following this approach, the Poynting vector of the element should be oriented towards the imaging target region. The single-side adapted dipole antenna is a novel design that fulfills this requirement. The performance of this design as a transmit coil array element has been characterized by comparison with three other, more conventional designs using finite difference time domain (FDTD) simulations and B +1 measurements on a phantom. Results show that the B +1 level at the deeper regions is higher while maintaining relatively low SAR levels. Also, the B +1 field distribution is more symmetrical and more uniform, promising better image homogeneity. Eight radiative antennas have been combined into a belt like surface array for prostate imaging. T(1) -weighted (T1W) and T(2) -weighted (T2W) volunteer images are presented along with B +1 measurements to demonstrate the improved efficiency. PMID- 21630343 TI - The concordance of MRI and quantitative autoradiography estimates of the transvascular transfer rate constant of albumin in a rat brain tumor model. AB - The apparent forward transfer constant, K transa, for albumin was measured in 9L cerebral tumors in 15 rats. An MRI study using gadolinium-labeled bovine serum albumin was followed by terminal quantitative autoradiography (QAR) using radioiodinated serum albumin. Look-Locker MRI estimates of T(1) followed gadolinium-labeled bovine serum albumin blood and tissue concentration. QAR and MRI maps of K transa were coregistered, a region of interest (ROI) that included the tumor and its surround was selected, and the two estimates of K transa from the ROI on QAR and MRI maps were compared by either mean per animal ROI or on pixel-by-pixel data using a generalized estimating equation. An ROI analysis showed a moderate correlation between the two measures (r = 0.57, P = 0.026); pixel-by-pixel generalized estimating equation analysis concurred (r = 0.54, P < 0.0001). The estimates of QAR with MRI of last time points (e.g., 25 min) showed a moderate correlation (ROI r = 0.55, P < 0.035; generalized estimating equation r = 0.58, P < 0.0001). Differences between the QAR and MRI estimates of K transa did not differ from zero, but the MRI 25-min estimate was significantly lower than the QAR estimate. Thus, noninvasive MRI estimates of vascular permeability can serve as a surrogate for QAR measures. PMID- 21630344 TI - Quantitative analysis of transmural gradients in myocardial perfusion magnetic resonance images. AB - Conventional quantitative assessments of myocardial perfusion analyze the temporal relation between the arterial input function and the myocardial signal intensity curves, thereby neglecting the important spatial relation between the myocardial signal intensity curves. The new method presented in this article enables characterization of sub-endocardial to sub-epicardial gradients in myocardial perfusion based on a two dimensional, "gradientogram" representation, which displays the evolution of the transmural gradient in myocardial contrast uptake over time in all circumferential positions of the acquired images. Moreover, based on segmentation in these gradientograms, several new measurements that characterize transmural myocardial perfusion distribution over time are defined. In application to clinical image data, the new two-dimensional representations, as well as the newly defined measurements revealed a clear distinction between normal perfusion and inducible ischaemia. Thus, the new measurements may serve as diagnostic markers for the detection and characterization of epicardial coronary and microvascular disease. PMID- 21630345 TI - Quantitative myocardial perfusion imaging with a MR cold pressor test. AB - The response of myocardial blood flow to sympathetic stimulation with cold is modulated by endothelium-related factors. As endothelial dysfunction is an early step in patients with coronary artery disease, the aim of this study was to establish a cold pressor test (CPT) setting for quantitative analysis of myocardial perfusion in a MR scanner. First pass perfusion studies were performed in 10 healthy volunteers using a 1.5 T MR scanner with a multislice steady state free precession perfusion trueFISP sequence in prebolus technique (1 cc/4 cc gadobenate dimeglumine). MR-CPT was established using an over head ice-water bath of the left hand. First pass perfusion imaging was started after 1 min to assure an adequate stimulus followed by a second series after 15 min to evaluate the rest perfusion. After motion correction images were segmented with an adapted, automated tool, myocardial contours were determined. Perfusion was quantitatively evaluated after contamination and baseline correction by deconvolution with the arterial input function using an exponential function model as residuum. All data could be evaluated. Mean myocardial perfusion rose from 0.61 +/- 0.22 cc/g/min at rest to 1.15 +/- 0.34 cc/g/min under CPT. MR myocardial perfusion values show a comparable increase under CPT as published positron emission tomography data. Consequently, CPT for the presence of endothelial dysfunction is feasible in the MR environment. PMID- 21630346 TI - Toward individualized SAR models and in vivo validation. AB - The specific absorption rate (SAR) is a limiting constraint in sequence design for high-field MRI. SAR estimation is typically performed by numerical simulations using generic human body models. This entails an intrinsic uncertainty in present SAR prediction. This study first investigates the required detail of human body models in terms of spatial resolution and the number of soft tissue classes required, based on finite-differences time-domain simulations of a 3 T body coil. The numerical results indicate that a resolution of 5 mm is sufficient for local SAR estimation. Moreover, a differentiation between fatty tissues, water-rich tissues, and the lungs was found to be essential to represent eddy current paths inside the human body. This study then proposes a novel approach for generating individualized body models from whole-body water-fat separated MR data and applies it to volunteers. The SAR hotspots consistently occurred in the arms due to proximity to the body coil as well as in narrow regions of the muscles. An initial in vivo validation of the simulated fields in comparison with measured B(1)-field maps showed good qualitative and quantitative agreement. PMID- 21630347 TI - MRI of frozen tissue demonstrates a phase shift. AB - While temperature mapping is desired during cryosurgery for prostate cancer treatment, an effective approach for this purpose is still needed. We have demonstrated a phase shift with temperature in our in vivo canine experiments and ex vivo tissue sample experiments within the frozen tissue. The phase shift is much larger (~0.7 degrees / degrees C with an echo time of 0.1 ms at 0.5 T) in magnitude than that predicted by conventional proton resonant frequency shift (0.008 degrees / degrees C). It shows little dependence on the echo times used and thus is not due to a frequency change, although frequency-dependent phase shift has been observed near the frozen tissue. This phase shift varies monotonically with temperature within the frozen tissue and therefore may be potentially used as a novel temperature mapping approach in cryoablation applications. PMID- 21630348 TI - Assessment of early diastolic strain-velocity temporal relationships using spatial modulation of magnetization with polarity alternating velocity encoding (SPAMM-PAV). AB - A novel MR imaging technique, spatial modulation of magnetization with polarity alternating velocity encoding (SPAMM-PAV), is presented to simultaneously examine the left ventricular early diastolic temporal relationships between myocardial deformation and intra-cavity hemodynamics with a high temporal resolution of 14 ms. This approach is initially evaluated in a dynamic flow and tissue mimicking phantom. A comparison of regional longitudinal strains and intra-cavity pressure differences (integration of computed in-plane pressure gradients within a selected region) in relation to mitral valve inflow velocities is performed in eight normal volunteers. Our results demonstrate that apical regions have higher strain rates (0.145 +/- 0.005 %/ms) during the acceleration period of rapid filling compared to mid-ventricular (0.114 +/- 0.007 %/ms) and basal regions (0.088 +/- 0.009 %/ms), and apical strain curves plateau at peak mitral inflow velocity. This pattern is reversed during the deceleration period, when the strain-rates in the basal regions are the highest (0.027 +/- 0.003 %/ms) due to ongoing basal stretching. A positive base-to-apex gradient in peak pressure difference is observed during acceleration, followed by a negative base-to-apex gradient during deceleration. These studies shed insight into the regional volumetric and pressure difference changes in the left ventricle during early diastolic filling. PMID- 21630349 TI - Myocardial BOLD imaging at 3 T using quantitative T2: application in a myocardial infarct model. AB - Left ventricular remodeling as a result of acute myocardial infarction (AMI) is associated with significant morbidity, leading to cardiovascular dysfunction, disability, and death. Despite successful revascularization, coronary vasodilatory dysfunction has been shown in infarcted and remote myocardium of patients following AMI. Our study explored the utility of a T(2)-based blood oxygen-level-dependent approach in probing regional and longitudinal fluctuations in vasodilatory function in a porcine model of AMI at 3 T. Ten pigs underwent MRI in control state and at day 2, weeks 1-6 following 90 min occlusion followed by reperfusion. The remote myocardium exhibited vasodilatory dysfunction at weeks 1 and 2 that gradually recovered, whereas the infarct zone showed no vasodilatory alterations. Our study suggests that microvascular alterations occurring in infarcted and remote myocardium after AMI might serve as an indicator of adverse left ventricular remodeling. The blood-oxygen-level-dependent technique using quantitative T(2) could potentially be a useful noninvasive tool to evaluate novel therapeutic strategies aimed at limiting vasoconstriction and improving coronary flow reserve after AMI. PMID- 21630350 TI - T2 -weighted MRI of post-infarct myocardial edema in mice. AB - T(2) -weighted, cardiac magnetic resonance imaging (T(2) w CMR) can be used to noninvasively detect and quantify the edematous region that corresponds to the area at risk (AAR) following myocardial infarction (MI). Previously, CMR has been used to examine structure and function in mice, expediting the study of genetic manipulations. To date, CMR has not been applied to imaging of post-MI AAR in mice. We developed a whole-heart, T(2) w CMR sequence to quantify the AAR in mouse models of ischemia and infarction. The DeltaB(0) and DeltaB(1) environment around the mouse heart at 7 T were measured, and a T(2) -preparation sequence suitable for these conditions was developed. Both in vivo T(2) w and late gadolinium enhanced CMR were performed in mice after 20-min coronary occlusions, resulting in measurements of AAR size of 32.5 +/- 3.1 (mean +/- SEM)% left ventricular mass, and MI size of 50.1 +/- 6.4% AAR size. Excellent interobserver agreement and agreement with histology were also found. This T(2) w imaging method for mice may allow for future investigations of genetic manipulations and novel therapies affecting the AAR and salvaged myocardium following reperfused MI. PMID- 21630351 TI - Closed circuit MR compatible pulsatile pump system using a ventricular assist device and pressure control unit. AB - The aim of this study was to evaluate the performance of a closed circuit MR compatible pneumatically driven pump system using a ventricular assist device as pulsatile flow pump for in vitro 3D flow simulation. Additionally, a pressure control unit was integrated into the flow circuit. The performance of the pump system and its test-retest reliability was evaluated using a stenosis phantom (60% lumen narrowing). Bland-Altman analysis revealed a good test-retest reliability (mean differences = -0.016 m/s, limits of agreement = +/-0.047 m/s) for in vitro flow measurements. Furthermore, a rapid prototyping in vitro model of a normal thoracic aorta was integrated into the flow circuit for a direct comparison of flow characteristics with in vivo data in the same subject. The pneumatically driven ventricular assist device was attached to the ascending aorta of the in vitro model to simulate the beating left ventricle. In the descending part of the healthy aorta a flexible stenosis was integrated to model an aortic coarctation. In vivo and in vitro comparison showed significant (P = 0.002) correlations (r = 0.9) of mean velocities. The simulation of increasing coarctation grade led to expected changes in the flow patterns such as jet flow in the post-stenotic region and increased velocities. PMID- 21630352 TI - Nonexponential T2 decay in white matter. AB - Visualizing myelin in human brain may help the study of diseases such as multiple sclerosis. Previous studies based on T(1) and T(2) relaxation contrast have suggested the presence of a distinct water pool that may report directly on local myelin content. Recent work indicates that T(2) contrast may offer particular advantages over T(1) and T(2) contrast, especially at high field. However, the complex mechanism underlying T(2) relaxation may render interpretation difficult. To address this issue, T(2) relaxation behavior in human brain was studied at 3 and 7 T. Multiple gradient echoes covering most of the decay curve were analyzed for deviations from mono-exponential behavior. The data confirm the previous finding of a distinct rapidly relaxing signal component (T(2) ~ 6 ms), tentatively attributed to myelin water. However, in extension to previous findings, this rapidly relaxing component displayed a substantial resonance frequency shift, reaching 36 Hz in the corpus callosum at 7 T. The component's fractional amplitude and frequency shift appeared to depend on both field strength and fiber orientation, consistent with a mechanism originating from magnetic susceptibility effects. The findings suggest that T(2) contrast at high field may be uniquely sensitive to tissue myelin content and that proper interpretation will require modeling of susceptibility-induced resonance frequency shifts. PMID- 21630353 TI - Fragile X-associated tremor ataxia syndrome sine tremor. PMID- 21630354 TI - Incidence, prescription patterns, and determinants of antipsychotic use in patients with Parkinson's disease. AB - Empirical data regarding the choice of antipsychotics for the management of psychosis in patients with Parkinson's disease are limited. This study aimed to evaluate the incidence and prescribing patterns of antipsychotics and to determine the predictors associated with the prescribing of typical antipsychotics in patients with Parkinson's disease. This was a retrospective cohort study analyzing data from the National Health Insurance Research Database in Taiwan between January 1, 2000, and December 31, 2006, in which patients with Parkinson's disease (ICD-9-CM codes 332) initially receiving any antiparkinsonian drug (n = 2095) were followed up to evaluate the subsequent use of antipsychotics. Kaplan-Meier statistics and multiple logistic regression were employed to evaluate the cumulative probability of antipsychotic use and determinants of prescribing of typical antipsychotics, respectively. The cumulative probability of initiation of an antipsychotic within 6 years was found to be 51%, and the proportion of patients who began taking an atypical antipsychotic increased from 11.1% in 2001 to 36.1% in 2005. Physician specialty was found to be the most influential predictor of the prescribing of typical antipsychotics: physicians with an internal medicine specialty were 10.62 times more likely (95% confidence interval, 4.64-24.32) to prescribe typical antipsychotics than were neurologists. The use of antipsychotics in Parkinson's disease is common, and the use of typical antipsychotics dominates antipsychotic treatment. Particular attention needs to be paid to improving practice, including efforts that encourage primary care providers to have the appropriate choice of antipsychotics in patients with Parkinson's disease. PMID- 21630355 TI - Inhibitory control during smooth pursuit in Parkinson's disease and Huntington's disease. AB - The basal ganglia are involved in the preferential selection and suppression of competing responses. Parkinson's disease and Huntington's disease are 2 prototypical basal ganglia disorders that feature impaired inhibitory control, a function of poor conflict resolution. Previous saccadic studies showed that individuals with Parkinson's disease experience difficulty suppressing unwanted ocular motor responses, whereas evidence for a similar difficulty in Huntington's disease is more equivocal. Relative to saccades, few research studies have examined inhibitory control processes in the context of an ongoing smooth pursuit task. In this study, we examined the ability of 16 patients with Parkinson's disease and 12 patients with Huntington's disease to suppress automatic responses to irrelevant distracters that transiently appeared during the tracking of a moving visual stimulus. Compared with an equivalent number of age-matched controls, patients with Parkinson's disease generated proportionately more saccades to distracter stimuli. This was particularly evident for distracters appearing far away from the target. Conversely, whereas individuals with early stage Huntington's disease and healthy controls made a comparable number of errors toward distracter stimuli, those in a more advanced clinical stage demonstrated significantly poorer inhibitory control. The current findings in parkinsonian patients replicate those previously reported in the saccadic and manual response literature, demonstrating difficulty inhibiting a competing motor response. However, in Huntington's disease we demonstrate for the first time that inhibitory control declines in more advanced-disease stages. This suggests that ocular motility may provide a sensitive marker of clinical disease progression in Huntington's disease. PMID- 21630356 TI - Atypical symptomatology of myoclonus dystonia (DYT-11) with positive response to bilateral pallidal deep brain stimulation. PMID- 21630357 TI - A missense mutation in CLIC2 associated with intellectual disability is predicted by in silico modeling to affect protein stability and dynamics. AB - Large-scale next generation resequencing of X chromosome genes identified a missense mutation in the CLIC2 gene on Xq28 in a male with X-linked intellectual disability (XLID) and not found in healthy individuals. At the same time, numerous nsSNPs (nonsynonomous SNP) have been reported in the CLIC2 gene in healthy individuals indicating that the CLIC2 protein can tolerate amino acid substitutions and be fully functional. To test the possibility that p.H101Q is a disease-causing mutation, we performed in silico simulations to calculate the effects of the p.H101Q mutation on CLIC2 stability, dynamics, and ionization states while comparing the effects obtained for presumably harmless nsSNPs. It was found that p.H101Q, in contrast with other nsSNPs, (a) lessens the flexibility of the joint loop which is important for the normal function of CLIC2, (b) makes the overall 3D structure of CLIC2 more stable and thus reduces the possibility of the large conformational change expected to occur when CLIC2 moves from a soluble to membrane form, and (c) removes the positively charged residue, H101, which may be important for the membrane association of CLIC2. The results of in silico modeling, in conjunction with the polymorphism analysis, suggest that p.H101Q may be a disease-causing mutation, the first one suggested in the CLIC family. PMID- 21630358 TI - Bootstrap estimation of confidence intervals on mutation rate ratios. AB - The fluctuation test is a useful tool for estimating the mutation rate of cells. However, statistical methods for comparing mutation rate estimates between different strains or conditions have not yet been fully developed. Methods exist for placing confidence intervals on estimates of the number of mutational events in cultures for a given strain and set of conditions, but placing confidence intervals on mutation rate is complicated by differences in the final number of cells in culture between parallel cultures. Additionally, confidence intervals on individual mutation rate estimates are not always the most useful statistical tool when comparing two or more different strains or conditions. We present a bootstrap method for estimating confidence intervals on the quotient of two mutation rates determined from two fluctuation test experiments using experimental and control strains or conditions. We use Monte Carlo simulations to validate this method over a wide range of mutation rates and for empirically measured variation in the estimates of final number of cells in culture. Furthermore, we provide the computational tools to implement the bootstrap method described here on experimental fluctuation test data and to evaluate this method for experimental parameters other than those considered herein. PMID- 21630359 TI - Identification of mutagens in freshwater sediments by the Ames-fluctuation assay using nitroreductase and acetyltransferase overproducing test strains. AB - Extracts of sediments from an area of concern in the Elbe river basins (Spittelwasser creek) were analyzed with the Ames-fluctuation test and in parallel with gas chromatography/mass spectrometry for compound identification. The standard test strains TA 98 and TA 100 showed mutagenicity mainly in medium polar fractions of the sediment extracts. PAHs contribute to the overall mutagenic potential of the sample. Especially, cyclopenta[c,d]pyrene that was previously not defined as a priority hazardous substance has to be considered as well. The addition of metabolically competent test strains, which overexpress nitroreductase and acetyltransferase (e.g., YG1041 and YG1042) to the test battery, increased significantly the sensitivity of the Ames test for medium polar to polar genotoxins. The increased mutagenicity that was found in these bacterial strains indicates the presence of nitroarenes and/or aromatic amines. In fact, a number of heterocyclic and nitrogen-substituted aromatic compounds were identified in the sediments of the Spittelwasser creek of which methyl parathion, 1-naphthylamine, and N-phenyl-2-naphthylamine are mutagenic. PMID- 21630360 TI - Protection and promotion of UV radiation-induced liposome leakage via DNA directed assembly with gold nanoparticles. PMID- 21630361 TI - Evaluation of the efficacy and safety of Salvia officinalis in controlling hot flashes in prostate cancer patients treated with androgen deprivation. AB - The purpose of this pilot study was to evaluate the efficacy and safety of Salvia officinalis in controlling hot flashes in prostate cancer patients treated with androgen deprivation therapy (ADT). Ten patients experiencing hot flashes were included in a single-centre prospective pilot study. Treatment consisted of 150 mg of Salvia officinalis extract taken orally three times daily. A diary questionnaire scoring hot flashes, subjective side effects and quality of life (QOL) had to be completed. Clinical examination was performed at every visit and the concentration of ADT-linked hormones, haemoglobin and cholesterol was measured before, during and after ending treatment. Before the start of treatment, a 1 week baseline registration was performed. An analysis of variance with time of measurement as a within-subject factor was performed. When analysing the hot flashes score, one patient was excluded due to insufficient diary notes. The mean weekly score declined from 112 (SD = 71) at baseline to 59 (SD = 54) at the end of treatment (p = 0.002). Hot flashes diminished significantly from the first week up to and including week 3. This was maintained during treatment. There was no effect on QOL. There were no side effects. It is concluded that Salvia officinalis is efficient and safe in the treatment hot flashes, without improving QOL. PMID- 21630362 TI - Agreement between two- and three-dimensional transperineal ultrasound methods in assessing fetal head descent in the first stage of labor. AB - OBJECTIVE: To study intraobserver repeatability and intermethod agreement between two- (2D) and three-dimensional (3D) transperineal ultrasound methods in assessing fetal head descent during the first stage of labor. METHODS: Fetal head descent was measured with transperineal ultrasound as the fetal head-perineum distance and the angle of progression in 106 primiparous women with prolonged first stage of labor. A single obstetrician performed all the scans, and another obstetrician analyzed the acquired 2D images and 3D volumes, blinded to clinical assessments and labor outcome. Intraobserver repeatability and intermethod agreement between 2D and 3D methods were analyzed. RESULTS: The repeatability coefficient was +/- 4.1 mm in 2D acquisitions and +/- 1.7 mm in 3D acquisitions of fetal head-perineum distance. The intraclass correlation coefficients (ICC) were 0.94 for 2D and 0.99 for 3D measurements. The angle of progression repeatability coefficients were +/- 6.7 degrees using 2D and +/- 5.7 degrees using 3D ultrasound and ICCs were 0.91 and 0.94, respectively. The intermethod ICC for fetal head-perineum distance in 2D vs 3D acquisitions was 0.95 and for angle of progression it was 0.93; the intermethod 95% limits of agreement were - 5.8 mm to + 7.2 mm and - 8.9 degrees to + 13.7 degrees , respectively. Cohen's kappa for 2D vs 3D acquisitions was 0.85 using 40 mm as a cut-off level for fetal head-perineum distance and 0.79 using 110 degrees as cut-off level for angle of progression. CONCLUSIONS: For one ultrasound operator the intraobserver repeatability and agreement between 2D and 3D ultrasound methods in prolonged first stage of labor were good. Given that 2D methods are simpler to learn and can be analyzed quickly online, 2D equipment might therefore be preferred in the labor room. PMID- 21630363 TI - Association of fetal cranial shape with shoulder dystocia. AB - OBJECTIVE: To evaluate whether fetal cranial shape is related to shoulder dystocia. METHODS: We compared shoulder dystocia cases (n = 18) with controls (normal vaginal deliveries, n = 18) in a retrospective matched-pairs observational study. Subjects were matched for known maternal and fetal risk factors and then evaluated for fetal biometric differences, which were measured by ultrasound near delivery. We tested multivariable risk models to predict shoulder dystocia by logistic regression. RESULTS: Cases had a smaller estimated occipitofrontal diameter (OFD) (P = 0.02) and a larger biparietal diameter/estimated OFD ratio (P = 0.003). A multivariable model including estimated fetal weight, estimated OFD, maternal weight and diabetes mellitus had sensitivity and specificity of 86% and 95%, respectively, and positive and negative likelihood ratios of 18.9 and 0.15, respectively. Estimated OFD significantly increased the predictive value of the model. CONCLUSION: A small estimated OFD is a risk factor for shoulder dystocia in the presence of other significant risk factors. A multivariable model including estimated OFD can predict shoulder dystocia in a clinically useful range. PMID- 21630364 TI - Three-dimensional sonography of the posterior fossa in fetuses with open spina bifida at 11-13 weeks' gestation. AB - OBJECTIVES: To investigate the posterior fossa of normal fetuses and fetuses with open spina bifida in stored three-dimensional (3D) volumes and to describe signs that might allow early detection of this defect. METHODS: A prospective study of 3D volumes of the fetal brain obtained from 10 normal fetuses and three fetuses with open spina bifida was undertaken. Measurements of the anteroposterior diameters of the cisterna magna and fourth ventricle were taken in the tilted axial view. In the mid-sagittal plane the brainstem (BS) diameter and the brainstem-occipital bone (BSOB) distance were measured. The BS/BSOB ratio was calculated. All measurements were expressed as Z-scores. Structural analysis of the differences in the posterior fossa between normal fetuses and fetuses with open spina bifida was undertaken. RESULTS: In normal fetuses all measurements were within +/-2.5 Z-scores. In three fetuses with open spina bifida the BS Z scores were 2.7, 2.8 and 2.8; the BSOB scores were -3.4, -2.8 and -2.9; the cisterna magna scores were -5.6, -3.7 and -4.2; and the BS/BSOB ratio scores were 4.1, 9.7 and 8.9. In normal fetuses the cisterna magna was posterior to the fourth ventricle and extended along its entire length. In fetuses with open spina bifida the cisterna magna was partially or completely obliterated. CONCLUSIONS: Assessment of the cranial posterior fossa is feasible at 11-13 weeks' gestation. There are distinct signs in fetuses with open spina bifida which can be evaluated by ultrasonography. PMID- 21630365 TI - Iris texture traits show associations with iris color and genomic ancestry. AB - OBJECTIVES: This study seeks to identify associations among genomic biogeographic ancestry (BGA), quantitative iris color, and iris texture traits contributing to population-level variation in these phenotypes. METHODS: DNA and iris photographs were collected from 300 individuals across three variably admixed populations (Portugal, Brazil, and Cape Verde). Two raters scored the photos for pigmentation spots, Fuchs' crypts, contraction furrows, and Wolflinn nodes. Iris color was quantified from RGB values. Maximum likelihood estimates of individual BGA were calculated from 176 ancestry informative markers. RESULTS: Pigmentation spots, Fuchs' crypts, contraction furrows, and iris color show significant positive correlation with increasing European BGA. Only contraction furrows are correlated with iris color. CONCLUSIONS: The relationship between BGA and iris texture illustrates a genetic contribution to this population-level variation. PMID- 21630366 TI - Perinatal winter conditions affect later reproductive performance in Romanian women: intra and intergenerational effects. AB - OBJECTIVES: To compare epidemiological studies on Austrian and New Zealand women that report below average reproductive performance after birth in summer, with a similar study in Romanian women. METHODS: We examined the association between birth month and offspring count analyzing census data from Romania obtained from IPUMS International (Population and Housing Census of Romania, year 2002), totaling 411,270 women aged older than 45 years. We investigated whether socio economic status affected this association, and whether intergenerational effects exist for mother's birth month in relation to their daughter's offspring number. RESULTS: The time series of mean offspring count per birth month has a highly significant period of 12 months. Contrary to our expectations, average offspring count is highest in women born in June and lowest in those born in December, with a strong coincidence between negative peak values of residuals of mean offspring count per birth month and corresponding monthly means of ambient temperature in Romania. The association between winter birth and lower offspring number is significant in poorly educated women only. For mothers born in winter there is also an association with a lower daughters' offspring count. CONCLUSIONS: Conflicting results exist between Romania and Austria/New Zealand, and may be most easily explained by women's different exposure to winter conditions in these countries. PMID- 21630367 TI - Testosterone and romance: the association of testosterone with relationship commitment and satisfaction in heterosexual men and women. AB - OBJECTIVES: The current study extends previous research on testosterone (T) and mating effort by examining whether relationship commitment and satisfaction explain variance in T beyond relationship status alone. METHODS: Salivary testosterone and self-reported assessments of relationship commitment and satisfaction were assessed among 90 heterosexual men and women (age M = 23.57) in a cross-sectional community sample. RESULTS: Relationship commitment was significantly related to T among men (P < 0.01), with increasing levels of commitment predicting lower T, even among paired men (P < 0.05). In contrast, relationship commitment was not related to women's T (P > 0.05). Controlling for relationship commitment, satisfaction did not predict T levels in men or women (P's > 0.18). CONCLUSIONS: The association of increasing relationship commitment with reduced T levels in men confirms and extends prior research linking T with mating effort. Together with previous research, this study suggests that T does not vary with relationship commitment or quality in monogamous, heterosexual women. PMID- 21630368 TI - Hand-washing, subclinical infections, and growth: a longitudinal evaluation of an intervention in Nepali slums. AB - OBJECTIVE: We conducted a longitudinal study to assess the impact of a hand washing intervention on growth and biomarkers of child health in Nepali slums. This is the first study to evaluate the impact of hand-washing on markers of subclinical, asymptomatic infections associated with childhood growth faltering. METHODS: We recruited a total sample of infants in the target age-range (3-12 months) living in the eight largest Kathmandu slums, allocating them to intervention (n = 45) and control (n = 43) groups. In intervention areas, a small scale community-based hand-washing program was implemented for six months; in control areas, mothers continued their normal practices. Time series linear regression was used to assess the impact of the intervention on levels of morbidity, mucosal damage, immune stimulation and growth. RESULTS: As expected, children with higher levels of mucosal damage exhibited worse growth over the period of the intervention (P = 0.01, <0.001 and 0.03 for height-for-age, weight for-age, and weight-for-height z-scores, respectively). We observed a 41% reduction in diarrheal morbidity (P = 0.023) for the intervention group relative to control. However, the hand-washing intervention did not lower levels of mucosal damage or immune stimulation, nor slow growth faltering. CONCLUSIONS: Reducing exposure to pathogens is an important global health priority. This study confirms the importance of hand-washing campaigns for reducing childhood morbidity. Yet our data suggest that promoting hand-washing is necessary but not sufficient to address chronic, subclinical infections. From a human biology standpoint, tackling the root causes of childhood infections is needed to address growth faltering in the context of highly contaminated slum environments. PMID- 21630369 TI - Genetics of head circumference in infancy: a longitudinal study of Japanese twins. AB - OBJECTIVES: Previous studies have shown strong genetic influence to head circumference (HC), but still little is known on the development of genetic etiology of HC in infancy, especially in non-Caucasian populations. Thus, we decided to analyze the genetics of HC growth in Japanese infants. METHODS: Longitudinal measures of HC were available from birth to 13 months of age in 206 monozygotic and 156 dizygotic complete twin pairs. Genetic modeling for twin data was used. RESULTS: We found only little evidence for sex-specific differences in the genetics of HC and thus analyzed boys and girls together. After 5 months of age the heritability of HC was high, but before that age also a substantial common environmental component was present. Not only strong genetic persistence for HC was found but also a new genetic variation emerged. New environmental variation shared by co-twins affecting HC was found until 3 months of age, and this effect was further transmitted until 1 year of age. CONCLUSIONS: HC and its growth are strongly genetically regulated. Largely, the same genetic factors affect the variation of HC at different ages, and new genetic variation emerged during the first year of life. Knowledge on the genetic component in the variation of HC may help to design tools for defining abnormal growth of HC in population-based screenings for related disorders. PMID- 21630370 TI - Growing into obesity: patterns of height growth in those who become normal weight, overweight, or obese as young adults. AB - OBJECTIVES: To study whether patterns of height growth differ by adult obesity status, and determine the contribution of subcutaneous fatness as an explanatory variable for any differences. STUDY DESIGN: A multicenter, prospective longitudinal cohort assessed in 3rd grade (8.8 years), 5th grade (11.1 years), 8th grade (14.1 years), and 12th grade (18.3 years). Exposures were young adult obesity status classified by CDC adult BMI categories at 12th grade. Skinfolds were measured in third, fifth, and eighth grades. Outcome was mean height (cm) at the four measurements using repeated-measures ANCOVA for young adult obesity status, and height increments between grades by adult obesity status in sequential models including initial height and, secondarily, initial skinfolds. RESULTS: Adjusted for age, and race/ethnicity, young adult obesity status explained a small, but statistically significant amount of height growth among both females and males within each of the three intervals. Compared with normal weight young adults, overweight or obese young adults stood taller in childhood, but had relatively less growth in height throughout the teenage years. There was no association between adult height and weight status. Skinfolds explained only a small amount of the height patterns in the three weight groups. CONCLUSION: Childhood and adolescent height growth patterns differ between those who become young adults who are normal weight and those who become overweight or obese. Since differences in fatness explain only a small amount of these height growth patterns, research is needed to identify other determinants. PMID- 21630371 TI - Frozen chicken for wild fish: nutritional transition in the Brazilian Amazon region determined by carbon and nitrogen stable isotope ratios in fingernails. AB - OBJECTIVES: Amazonian populations are experiencing dietary changes characteristic of the nutrition transition. However, the degree of change appears to vary between urban and rural settings. To investigate this process, we determined carbon and nitrogen stable isotope ratios in fingernails and dietary intake of Amazonian populations living along a rural to urban continuum along the Solimoes River in Brazil. METHODS: Carbon and nitrogen stable isotope ratios were analyzed from the fingernails of 431 volunteer subjects living in different settings ranging from rural villages, small towns to urban centers along the Solimoes River. Data from 200 dietary intake surveys were also collected using food frequency questionnaires and 24-h recall interviews in an effort to determine qualitative aspects of diet composition. RESULTS: Fingernail delta(13) C values (mean +/- standard deviation) were -23.2 +/- 1.3, -20.2 +/- 1.5, and -17.4 +/- 1.30/00 and delta(15) N values were 11.8 +/- 0.6, 10.4 +/- 0.8, and 10.8 +/- 0.70/00 for those living in rural villages, small towns, and major cities, respectively. We found a gradual increase in the number of food items derived from C(4) plant types (meat and sugar) and the replacement of food items derived from C(3) plant types (fish and manioc flour) with increasing size of urban centers. CONCLUSION: Increasing urbanization in the Brazilian Amazon is associated with a significant change in food habits with processed and industrialized products playing an increasingly important role in the diet and contributing to the nutrition transition in the region. PMID- 21630372 TI - Sex differences in birth size and intergenerational effects of intrauterine exposure to Ramadan in Saudi Arabia. AB - OBJECTIVES: In Europe, boys and girls have different body proportions at birth. We examined newborn babies in Saudi Arabia to determine the sex differences and whether fetal growth differed if the mother was in utero during Ramadan. METHODS: We examined body size at birth among 967 babies (479 boys and 488 girls) born in Unizah, a small city in Saudi Arabia. RESULTS: Large head circumference was the strongest single predictor of male sex. In a simultaneous regression, female sex was predicted by small head circumference (P < 0.001), low birth weight (P = 0.002), and large chest circumference (P = 0.008). The mothers of boys were heavier in pregnancy than the mothers of girls and had a higher body mass index, 31.7 kg/m(2) compared to 30.2 (P < 0.001). The mothers of girls, however, were taller than the mothers of boys, 158.6 cm compared to 157.4 (P = 0.001). Compared to babies whose mothers were not in utero during Ramadan boys whose mothers were in mid gestation during Ramadan were 1.2 cm longer (P = 0.005) while girls had a 0.4 week shorter gestation period (P = 0.04). CONCLUSION: Our findings are consistent with other evidence that boys are more ready than girls to trade off visceral development in utero to protect somatic and brain growth. They also support the hypothesis that boys are more responsive to their mother's current diet than girls, who respond more to their mother's life time nutrition and metabolism. They provide the first evidence that changes in the life style of pregnant women during Ramadan affect more than one generation. PMID- 21630373 TI - The absence of the calcium-buffering protein calbindin is associated with faster age-related decline in hippocampal metabolism. AB - Although reductions in the expression of the calcium-buffering proteins calbindin D-28K (CB) and parvalbumin (PV) have been observed in the aging brain, it is unknown whether these changes contribute to age-related hippocampal dysfunction. To address this issue, we measured basal hippocampal metabolism and hippocampal structure across the lifespan of C57BL/6J, calbindin D-28k knockout (CBKO) and parvalbumin knockout (PVKO) mice. Basal metabolism was estimated using steady state relative cerebral blood volume (rCBV), which is a variant of fMRI that provides the highest spatial resolution, optimal for the analysis of individual subregions of the hippocampal formation. We found that like primates, normal aging in C57BL/6J mice is characterized by an age-dependent decline in rCBV estimated dentate gyrus (DG) metabolism. Although abnormal hippocampal fMRI signals were observed in CBKO and PVKO mice, only CBKO mice showed accelerated age-dependent decline of rCBV-estimated metabolism in the DG. We also found age independent structural changes in CBKO mice, which included an enlarged hippocampus and neocortex as well as global brain hypertrophy. These metabolic and structural changes in CBKO mice correlated with a deficit in hippocampus dependent learning in the active place avoidance task. Our results suggest that the decrease in CB that occurs during normal aging is involved in age-related hippocampal metabolic decline. Our findings also illustrate the value of using multiple MRI techniques in transgenic mice to investigate mechanisms involved in the functional and structural changes that occur during aging. PMID- 21630374 TI - Complimentary roles of the hippocampus and retrosplenial cortex in behavioral context discrimination. AB - Complex cognitive functions, such as learning and memory, arise from the interaction of multiple brain regions that comprise functional circuits and different components of these circuits make unique contributions to learning. The hippocampus and the retrosplenial cortex (RSC) are anatomically interconnected and both regions are involved in learning and memory. Previous studies indicate that the hippocampus exhibits unique firing patterns for different contexts and that RSC neurons selectively respond to cues that predict reinforcement or the need for a behavioral response, suggesting a hippocampal role in encoding contexts and an RSC role in encoding behaviorally significant cues. To test this, we simultaneously recorded hippocampal and RSC neuronal activity as rats learned to discriminate two behavioral contexts. The rats learned to approach the east arm of a plus maze for reward during the first half of each session and to approach the west arm during the second half. The "go east" and "go west" conditions constitute distinct behavioral contexts, which were cued by the reward location. Neurons in both regions developed highly context-specific responses as subjects learned to discriminate the contexts, but the response patterns differed in the two brain regions. Consistent with a context processing role, hippocampal neurons developed context-specific responses to a variety of task stimuli and events. In contrast, RSC neurons only developed context-specific responses to the reward location, which served as the context identifying cue. These results suggest that the hippocampus and RSC play distinct, but complimentary roles in mediating context appropriate memories and behaviors. PMID- 21630375 TI - Assessing the impact of high deductible health plans on health-care utilization and cost: a changes-in-changes approach. AB - High deductible health plans (HDHPs) have become an increasingly common form of benefit design used by employers to manage health-care costs. Numerous studies have evaluated the uptake and impact of HDHPs on health-care utilization. Most studies have employed the standard difference-in-differences (DID) methodology. In this paper, we employ three alternative methodologies to evaluate a natural experiment in which a traditional health plan was fully replaced by a HDHP. We implement the standard DID and the quantile difference-in-differences (QDID) estimators to evaluate the impact of the HDHP on following six outcomes: overall cost, medical cost, pharmacy cost, outpatient visit count, inpatient visit count and emergency room visit count. We compare these results to a changes-in-changes (CIC) estimator, a generalized version of the standard (DID) estimator. We find that both the DID and CIC models yielded similar results, while the QDID model provided additional insights on the HDHP impact across different parts of the outcome distributions. Overall, introduction of HDHP had no impact on health-care costs, positive impact on the number of outpatient visits and mixed impacts on the inpatient and emergency room visit counts. The QDID estimates suggest HDHP introduction generally impacted subjects in upper percentiles (50th, 75th and 90th). PMID- 21630376 TI - A brain-potential study of preparation for and execution of a task-switch with stimuli that afford only the relevant task. AB - Behavioural and neurophysiological studies of task-switching have tended to employ 'bivalent' stimuli (which afford responses in two tasks). Using brain potential recordings, we investigated task-switching with 'univalent' stimuli affording responses in only one of the tasks, and compared the outcomes to those recently obtained with bivalent stimuli (Lavric et al. [2008]: Eur J Neurosci 1 14), in order to examine two phenomena. First, when only univalent stimuli are presented, the processing of task cues becomes optional. Our results showed that in these circumstances linguistic (but not pictorial) cues were still effective in eliciting at least some degree of preparation for a task-switch, as evidenced by the reduction in the error cost of switching at the longer preparation interval and by a posterior switch-induced ERP positivity at about 450-800 ms in the cue-stimulus interval. Second, single affordance stimuli not only reduced behavioural switch costs relative to bivalent stimuli; they also produced a smaller post-stimulus switch-induced negativity, consistent with the latter being a marker of conflict between task-sets. However, using stimuli not associated with responses in the alternative task did not completely eliminate the negativity. We speculate that the residue reflects other sources of conflict: attention to the irrelevant perceptual dimension and/or persistence of task goals. PMID- 21630377 TI - Activin/Nodal signaling controls divergent transcriptional networks in human embryonic stem cells and in endoderm progenitors. AB - Activin/Nodal signaling is necessary to maintain pluripotency of human embryonic stem cells (hESCs) and to induce their differentiation toward endoderm. However, the mechanisms by which Activin/Nodal signaling achieves these opposite functions remain unclear. To unravel these mechanisms, we examined the transcriptional network controlled in hESCs by Smad2 and Smad3, which represent the direct effectors of Activin/Nodal signaling. These analyses reveal that Smad2/3 participate in the control of the core transcriptional network characterizing pluripotency, which includes Oct-4, Nanog, FoxD3, Dppa4, Tert, Myc, and UTF1. In addition, similar experiments performed on endoderm cells confirm that a broad part of the transcriptional network directing differentiation is downstream of Smad2/3. Therefore, Activin/Nodal signaling appears to control divergent transcriptional networks in hESCs and in endoderm. Importantly, we observed an overlap between the transcriptional network downstream of Nanog and Smad2/3 in hESCs; whereas, functional studies showed that both factors cooperate to control the expression of pluripotency genes. Therefore, the effect of Activin/Nodal signaling on pluripotency and differentiation could be dictated by tissue specific Smad2/3 partners such as Nanog, explaining the mechanisms by which signaling pathways can orchestrate divergent cell fate decisions. PMID- 21630379 TI - CID of singly charged antioxidants applied in lubricants by means of a 3D ion trap and a linear ion trap-Orbitrap mass spectrometer. AB - The aim of this study was to investigate the fragmentation behavior induced by low-energy collision-induced dissociation (LE-CID) of four selected antioxidants applied in lubricants, by two different types of ion trap mass spectrometers: a three-dimensional ion trap (3D-IT) and a linear IT (LIT) Orbitrap MS. Two sterically hindered phenols and two aromatic amines were selected as model compounds representing different antioxidant classes and were characterized by positive-ion electrospray ionization (ESI) and LE-CID. Various types of molecular ions (e.g. [M](+*) , [M + H](+) , [M + NH(4) ](+) or [M + Na](+) ) were used as precursor ions generating a significant number of structurally relevant product ions. Furthermore, the phenolic compounds were analyzed by negative-ion ESI. For both IT types applied for fragmentation, the antioxidants exhibited the same unusual LE-CID behavior: (1) they formed stable radical product ions and (2) C?C bond cleavages of aliphatic substituents were observed and their respective cleavage sites depended on the precursor ion selected. This fragmentation provided information on the type of structural isomer usually not obtainable for branched aliphatic substituents utilizing LE-CID. Comparing the two instruments, the main benefit of applying the LIT-Orbitrap was direct access to elemental composition of product ions enabling unambiguous interpretation of fragmentation trees not obtainable by the 3D-IT device (e.g. loss of isobaric neutrals). It should be emphasized that the types of product ions formed do not depend on the type of IT analyzer applied. For characterizing degradation products of antioxidants, the LIT-Orbitrap hybrid system, allowing the determination of accurate m/z values for product ions, is the method of choice. PMID- 21630378 TI - Bone marrow stromal cells produce long-term pain relief in rat models of persistent pain. AB - Chronic pain conditions are difficult to treat and are major health problems. Bone marrow stromal cells (BMSCs) have generated considerable interest as a candidate for cell-based therapy. BMSCs are readily accessible and are easy to isolate and expand ex vivo. Clinical studies show that direct injection of BMSCs does not produce unwanted side effects and is well tolerated and safe. Here, we show that a single systemic (intravenous) or local injection (into the lesion site) of rat primary BMSCs reversed pain hypersensitivity in rats after injury and that the effect lasted until the conclusion of the study at 22 weeks. The pain hypersensitivity was rekindled by naloxone hydrochloride, an opioid receptor antagonist that acts peripherally and centrally, when tested at 1-5 weeks after BMSC infusion. In contrast, naloxone methiodide, a peripherally acting opioid receptor antagonist, only rekindled hyperalgesia in the first 3 weeks of BMSC treatment. Focal downregulation of brainstem mu opioid receptors by RNA interference (RNAi) reversed the effect of BMSCs, when RNAi was introduced at 5- but not 1-week after BMSC transplantation. Thus, BMSCs produced long-term relief of pain and this effect involved activation of peripheral and central opioid receptors in distinct time domains. The findings prompt studies to elucidate the cellular mechanisms of the BMSC-induced pain relieving effect and translate these observations into clinical settings. PMID- 21630380 TI - Proton transfer reactions for improved peptide characterisation. AB - The combination of deprotonation (via ion/molecule and ion/ion reactions) and low energy collision-induced dissociation (CID) has been explored for the enhanced characterisation of tryptic peptides via access to different precursor charge states. This approach allows instant access to fragmentation properties of singly and doubly protonated precursors (arising from the availability of mobile protons) in a single experiment. Considering both charge states extended our base of structurally informative data (in comparison with considering just a single charge state) due to generation of additional sequence ions and by obtaining supplementary structural information derived from selective cleavages. Roughly 37% of combined data sets (CID spectra of doubly and singly charged precursor) showed a greater database identification confidence than each set alone. Moreover, comparison between a number of sequence ions of the singly charged precursor and the doubly charged precursor provided a mean of distinguishing the two classes of tryptic peptides (arginine or lysine containing). PMID- 21630381 TI - Serine-phosphoric acid cluster ions studied by electrospray ionization and tandem mass spectrometry. AB - More than 310 kinds of cluster ions of S(m) P(n) H(k) (k+) are observed in a single ESI mass spectrum of a mixed solution of serine and phosphoric acid. Some typical cluster ions are selected, activated by collision in a FT ICR cell, and the dissociation pathways were deduced in detail. For large singly protonated ions, the collisions cause the ejection of subunits of serine or phosphoric acid subsequently producing the ions of S(2) P(4) H(1) (1+) , which can be further dissociated by the loss of phosphoric acid molecules in turn and form the protonated serine dimer and monomer. However, for the doubly protonated ions, the dissociation pathways change from the loss of a protonated serine dimer for the ions of S(7) P(9) H(2) (2+) to the neutral loss of H(3) PO(4) for the ions of S(7) P(12) H(2) (2+) or the neutral loss of serine or H(3) PO(4) for the larger clusters, indicating the effect of cluster sizes on the process of dissociation. The structure of S(2) P(4) H(1) (1+) is suggested based on B3LYP/6-31G(d,p) calculations. The diversity and structural orderliness of the hetero-cluster ions are mainly attributed to the network of hydrogen bonds inside the cluster ions and the extraordinary amphotericity of the components. PMID- 21630382 TI - A neutralization-reionization and reactivity mass spectrometry study of the generation of neutral hydroxymethylene. AB - Neutral hydroxymethylene HCOH is an important intermediate in several chemical reactions; however, it is difficult to observe due to its high reactivity. In this work, neutral hydroxymethylene and formaldehyde were generated by charge exchange neutralization of their respective ionic counterparts and then were reionized and detected as positive-ion recovery signals in neutralization reionization mass spectrometry in a magnetic sector instrument of BEE geometry. The reionized species were characterized by their subsequent collision-induced dissociation mass spectra. The transient hydroxymethylene neutral was observed to isomerize to formaldehyde with an experimental time span exceeding 13.9 us. The vertical neutralization energy of the HCOH(+*) ion has also been assayed using charge transfer reactions between the fast ions and stationary target gases of differing ionization energy. The measured values match the result of ab initio calculations at the QCISD/6-311 + G(d,p) and CCSD(T)/6-311 + + G(3df,2p) levels of theory. Neutral hydroxymethylene was also produced by proton transfer from CH(2) OH(+) to a strong base such as pyridine, confirmed by appropriate isotopic labeling. There is a kinetic isotope effect (KIE) for H(+) versus D(+) transfer from the C atom of the hydroxymethyl cation of ~3, consistent with a primary KIE of a nearly thermoneutral reaction. PMID- 21630383 TI - Characteristics and origins of common chemical noise ions in negative ESI LC-MS. AB - Ionic chemical background noise in LC-MS has been one of the major problems encountered in trace analysis. In this study, the typical negative background ions in ESI LC-MS are investigated exemplarily. It was carried out using tandem mass spectrometry to study the products and precursors of the major background ions to examine their structures and structure relationship. Various typical LC eluents with different compositions and additives such as ammonium formate/formic acid and ammonium acetate/acetic acid have been studied. Several types of negative noise ions are concluded, which include the cluster chemical background ions only from mobile phase components and additives. Furthermore, there are also abundant clusters resulting from the solvation of some typical individual contaminants (e.g. additives and degradation products from tubing, impurities in the mobile phase, etc.), accompanied by some minor contribution from contaminants. The elemental composition of some selected ions was confirmed using the FT-ICR accurate mass measurement. This work provides us insight into information about the structures and types of common negative background ions and will help to understand their formation and origins. More importantly, it will guide us to prevent chemical noise interference in practice and also contribute to develop methods for noise reduction based on selective ion-molecule reactions. PMID- 21630384 TI - Presence of galactosylated core fucose on N-glycans in the planaria Dugesia japonica. AB - Planarial species are of especial interest to biologists due to the phenomenon of pluripotency and, in comparison to other developmental processes, it can be hypothesised that glycan-lectin interactions may play a role. In order to examine the N-glycans of one of these organisms, Dugesia japonica, peptide:N-glycosidase A was employed and the released glycans were subject to pyridylamination, HPLC and mass spectrometric analysis. A range of oligomannosidic glycans was observed with a trimethylated Man(5) GlcNAc(2) structure being the dominant species. Three glycans were also observed to contain deoxyhexose; in particular, a glycan with the composition Hex(4) HexNAc(2) Fuc(1) Me(2) was revealed by exoglycosidase digestion, in combination with MS/MS, to contain a galactosylated core alpha1,6 fucose residue, whereas this core modification was found to be capped with a methylhexose residue in the case of a Hex(5) HexNAc(2) Fuc(1) Me(3) structure. This is the first report of these types of structures in a platyhelminth and indicates that the 'GalFuc' modification of N-glycans is not just restricted to molluscs and nematodes. PMID- 21630385 TI - High spatial resolution imaging mass spectrometry and classical histology on a single tissue section. AB - The work presented in this report describes and demonstrates a protocol for protein imaging analysis of biological tissue using MALDI IMS where histological staining and MS analysis are performed on the same tissue section. Spatial image resolution is shown at 35 um for sagittal sections of the cerebellum from rat brain. PMID- 21630386 TI - Gaseous cation chemistry and chain-length effects in electron ionization and collision-induced dissociation mass spectra of symmetric 1,n-bis(9 anthracenyl)alkanes. AB - The behavior of the gaseous cations resulting from EI (30 and 70 eV) of the bichromophoric title compounds 1-5 (for n = 1-5, respectively) is examined by ion trap mass spectrometry, including collision-induced dissociation (CID) with variation in collision energy. These results are compared with those from anthracene and 9-methylanthracene and with previously reported mass spectrometric results for 3 and dicarbazolylalkanes. Rather than using the kinetic method to obtain ion energetics where the fragmentation mechanism is clear, as commonly done, the method is used here with relative complementary-ion abundances from CID to test the proposed fragmentation mechanisms using B3LYP calculations of relative ionization energies and optimized geometries of ionic and neutral fragments. Hydrogen migrations are common, and skeletal rearrangements including formation of expanded, fused and spiro rings are proposed in several cases. Of the chain cleavages, alpha-homolysis giving C(15) H(11) (+) , likely as dibenzotropylium, is most important for each of 1-5 except 3, where beta-cleavage to C(16) H(13) (+) dominates with a proposed methyldibenzotropylium structure. alpha-Cleavage was important also in the dicarbazolylalkanes. A previous inference of a McLafferty rearrangement to explain C(15) H(12) (+*) from 3 is not supported by the present results. The fragmentation behavior of 1-5 depends strongly on n and implies significant interchromophoric interaction between anthracenyl groups. PMID- 21630387 TI - Thymine quintets and their higher order assemblies studied by electrospray ionization mass spectrometry and theoretical calculation. AB - We previously reported that thymine molecules can specifically form a pentameric magic number cluster named as thymine quintet in the presence of K(+) , Rb(+) and Cs(+) . Actually, thymine decamer and doubly charged thymine 15-mer metaclusters can be observed along with thymine quintet in the ESI mass spectra of thymine with the addition of K(+) , Rb(+) and Cs(+) . The product ion spectra of these metaclusters, especially the 15-mer with hetero central ions, indicate that they are higher order assemblies of thymine quintets. The collision-induced dissociation experiments show that the gas-phase stabilities of these metaclusters depend on the size of the central ions, following the order Cs(+) > Rb(+) > K(+) , while K(+) leads to the highest dissociation energy of a thymine quintet. The optimized structures of thymine quintet and decamer were provided by density functional theory calculations, which showed that thymine quintet is bowl shaped and its tilting angle increases with the size of the central ion. Furthermore, the chirality of thymine quintet was defined for the first time and the resulting different diastereoisomers of thymine decamers were also revealed by the calculation study. Copyright (c) 2011 John Wiley & Sons, Ltd. PMID- 21630388 TI - It is time for a paradigm shift in drug discovery bioanalysis: from SRM to HRMS. AB - It can be argued that the last true paradigm shift in the bioanalytical (BA) arena was the shift from high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection to HPLC with tandem mass spectrometry (MS/MS) detection after the commercialization of the triple quadrupole mass spectrometer in the 1990s. HPLC-MS/MS analysis based on selected reaction monitoring (SRM) has become the gold standard for BA assays and is used by all the major pharmaceutical companies for the quantitative analysis of new drug entities (NCEs) as part of the new drug discovery and development process. While LC-MS/MS continues to be the best tool for drug discovery bioanalysis, a new paradigm involving high-resolution mass spectrometry (HRMS) and ultrahigh-pressure liquid chromatography (uHPLC) is starting to make inroads into the pharmaceutical industry. The ability to collect full scan spectra, with excellent mass accuracy, mass resolution, 10-250 ms scan speeds and no NCE-related MS parameter optimization, makes the uHPLC-HRMS techniques suitable for quantitative analysis of NCEs while preserving maximum qualitative information about other drug-related and endogenous components such as metabolites, degradants, biomarkers and formulation materials. In this perspective article, we provide some insight into the evolution of the hybrid quadrupole-time-of-flight (Qq-TOF) mass spectrometer and propose some of the desirable specifications that such HRMS systems should have to be integrated into the drug discovery bioanalytical workflow for performing integrated qualitative and quantitative bioanalysis of drugs and related components. PMID- 21630389 TI - Ulcerative colitis complicated by atypical peristomal pyoderma gangrenosum. PMID- 21630390 TI - Stereo-specific inhibition of acetyl- and butyryl-cholinesterases by enantiomers of cis,cis-decahydro-2-naphthyl-N-n-butylcarbamate. AB - Enantiomers of cis,cis-decahydro-2-naphthyl-N-n-butylcarbamate show stereo specific inhibition for acetylcholinesterase and butyrylcholinesterase. For both inhibition reaction, (2S,4aR,8aS)-cis,cis-decahydro-2-naphthyl-N-n- butylcarbamate is more potent than (2R,4aS,8aR)-cis,cis-decahydro-2-naphthyl-N-n butylcarbamate. Optically pure (2S,4aR,8aS)-(-)- and (2R,4aS,8aR)-(+)-cis,cis decahydro-2-naphthols are resolved by the porcine pancreatic lipase-catalyzed acetylation of decahydro-2-naphthols with vinyl acetate. Absolute configurations and the enantiomeric excess values of (2S,4aR,8aS)-(-)- and (2R,4aS,8aR)-(+) cis,cis-decahydro-2-naphthols are determined from the (19)F NMR spectra of their Mosher's ester derivatives. We fail to resolve (2S,4aR,8aR)- and (2R,4aS,8aS) trans,cis-decahydro-2-naphthols from the porcine pancreatic lipase-catalyzed acetylation of decahydro-2-naphthols with vinyl acetate. PMID- 21630391 TI - Antioxidant role of coumarin on streptozotocin-nicotinamide-induced type 2 diabetic rats. AB - The purpose of this study was to investigate the antioxidant role of coumarin on streptozotocin-nicotinamide-induced Type 2 diabetic rats. In experimental rats, the levels of plasma glucose, insulin, and the levels of thiobarbituric acid reactive substances, lipid hydroperoxides, conjugated dienes, and the activities of superoxide dismutase, catalase, glutathione-S-transferase, and glutathione peroxidase were assayed in liver and kidney. Diabetic rats showed elevated levels of plasma glucose and lipid peroxidation markers and reduced plasma insulin and antioxidant enzymes. Oral administration of coumarin resulted in a significant reduction in the plasma glucose and lipid peroxides and a significant increase in the plasma insulin and antioxidant enzymes. Chronic treatment of coumarin remarkably restored the normal status of the histopathological changes observed in the selected tissues. It can be concluded that coumarin has antioxidant effect in Type 2 diabetic rats. PMID- 21630394 TI - An oxygen-deficient perovskite as selective catalyst in the oxidation of alkyl benzenes. PMID- 21630395 TI - Palladium-catalyzed bromoalkynylation of C-C double bonds: ring-structure dependent synthesis of 7-alkynyl norbornanes and cyclobutenyl halides. PMID- 21630396 TI - The influence of hydrogen-bond defects on the properties of ionic liquids. PMID- 21630397 TI - A palladium-catalyzed multicomponent coupling approach to pi-conjugated oligomers: assembling imidazole-based materials from imines and acyl chlorides. PMID- 21630398 TI - The mechanism of the titanium-catalyzed hydroaminoalkylation of alkenes. PMID- 21630399 TI - Fragment condensation of C-terminal pseudoproline peptides without racemization on the solid phase. PMID- 21630400 TI - A molecular four-stroke motor. PMID- 21630401 TI - Low-temperature catalytic H2 oxidation over Au nanoparticle/TiO2 dual perimeter sites. PMID- 21630402 TI - Total synthesis of the analgesic conotoxin MrVIB through selenocysteine-assisted folding. PMID- 21630403 TI - Extension of the Stober method to the preparation of monodisperse resorcinol formaldehyde resin polymer and carbon spheres. PMID- 21630404 TI - A facile palladium-mediated contraction of benzene to cyclopentadiene: transformations of palladium(II) p-benziporphyrin. PMID- 21630405 TI - Highly enantioselective catalytic synthesis of functionalized chiral diazoacetoacetates. PMID- 21630406 TI - Enantioselective synthesis of siloxyallenes from alkynoylsilanes by reduction and a Brook rearrangement and their subsequent trapping in a [4+2] cycloaddition. PMID- 21630408 TI - Switching modulation for protein labeling with activatable fluorescent probes. PMID- 21630409 TI - Dissecting the role of single regions of an IAPP mimic and IAPP in inhibition of Abeta40 amyloid formation and cytotoxicity. AB - Alzheimer's disease (AD) and type 2 diabetes (T2D) are linked to the self association of beta-amyloid peptide (Abeta) and islet amyloid polypeptide (IAPP), respectively. We have shown that IAPP-GI, a soluble IAPP analogue and mimic of nonamyloidogenic and nontoxic IAPP, binds Abeta with high affinity and blocks its cytotoxic self-assembly and fibrillogenesis. We have also shown that IAPP and Abeta interact with each other into nonfibrillar and nontoxic heterocomplexes that suppress cytotoxic self-association by both polypeptides. The Abeta-IAPP interaction might thus be a molecular link between AD and T2D. We studied the role of individual IAPP-GI and IAPP regions in their inhibitory function on Abeta40 self-association and cytotoxicity. We found that the presence of the two hot-spot regions of the Abeta-IAPP interaction interface in IAPP(8-28) is not sufficient for inhibitory function and that, in addition to IAPP(8-28), the presence of the N-terminal region IAPP(1-7) is absolutely required. By contrast, the C-terminal region, IAPP(30-37), is not required although its presence together with IAPP(1-7) in IAPP-GI results in a marked enhancement of the inhibitory effect as compared to IAPP(1-28)-GI. We suggest that the inhibitory effect of IAPP-GI and IAPP on Abeta40 fibrillogenesis and cell toxicity is mediated primarily by interactions involving the hot regions of the Abeta-IAPP interaction interface and the N terminus of IAPP while a concerted and likely structure-stabilizing action of the N- and C-terminal IAPP regions potentiates this effect. These results identify important molecular determinants of the amyloid suppressing function of the Abeta40-IAPP interaction and could contribute to the design of novel inhibitors of Abeta40 aggregation and cell degeneration. PMID- 21630410 TI - Protein containers--promising tools for the future. PMID- 21630411 TI - Alternating current measurements in scanning electrochemical microscopy, part 2: detection of adsorbates. AB - A scanning electrochemical microscope (SECM) in ac mode is used for the characterisation of the adsorption process during the hydrogen evolution reaction (HER) in sulfuric acid solution. It is shown that this technique allows quantitative analysis of the adsorption process, and measurements of the differential capacitance with the frequency as parameter are obtained. The time constant for relaxation of adsorbed hydrogen (H(ads)) is approximately 2 Hz, and analysis of the Nyquist plot allows direct evaluation of the charge involved. In addition, the direct comparison of the usual electrochemical impedance data and ac-SECM results obtained simultaneously permits characterisation of processes occurring at the surface and in solution. PMID- 21630412 TI - Improved photon yield from a green dye with a reducing and oxidizing system. PMID- 21630413 TI - Dynamic enantiodivergent organocatalysis: merging molecular motors with bifunctional organocatalysts. PMID- 21630414 TI - Genome-wide association studies (GWAS): impact on elucidating the aetiology of diabetes. AB - It has proven to be challenging to isolate the genes underlying the genetic components conferring susceptibility to type 1 and type 2 diabetes. Unlike previous approaches, 'genome-wide association studies' have extensively delivered on the promise of uncovering genetic determinants of complex diseases, with a number of novel disease-associated variants being largely replicated by independent groups. This review provides an overview of these recent breakthroughs in the context of type 1 and type 2 diabetes, and outlines strategies on how these findings will be applied to impact clinical care for these two highly prevalent disorders. PMID- 21630415 TI - Generation of transgenic dogs that conditionally express green fluorescent protein. AB - We report the creation of a transgenic dog that conditionally expresses eGFP (enhanced green fluorescent protein) under the regulation of doxycycline. Briefly, fetal fibroblasts infected with a Tet-on eGFP vector were used for somatic cell nuclear transfer. Subsequently reconstructed oocytes were transferred to recipients. Three clones having transgenes were born and one dog was alive. The dog showed all features of inducible expression of eGFP upon doxycycline administration, and successful breeding resulted in eGFP-positive puppies, confirming stable insertion of the transgene into the genome. This inducible dog model will be useful for a variety of medical research studies. PMID- 21630416 TI - Production of offspring from cloned transgenic RFP female dogs and stable generational transmission of the RFP gene. AB - The purpose of this study was to analyze the reproductive ability of transgenic female dogs born bysomatic cell nuclear transfer and to determine inheritance of the red fluorescent protein (RFP) transgene. The four founder transgenic bitches (F0) reached puberty at 340.8 +/- 39.6 days after birth and were bred with wild type male dogs by natural mating or by artificial insemination. The bitches all became pregnant and successfully delivered 13 puppies (F1), of which two females were bred with wild-type dogs to deliver 7 offspring (F2), including 1 stillbirth. Among the 19 live offspring, 10 puppies showed emission of RFP under UV light and the presence of the RFP transgene was confirmed by genomic PCR and Southern blot analyses. In conclusion, transgenic RFP female dogs exhibited normal reproductive ability and expression of the transgene was demonstrated in F1 and F2 generations. PMID- 21630420 TI - Cellular DNA methylation program during neurulation and its alteration by alcohol exposure. AB - BACKGROUND: Epigenetic changes are believed to be among the earliest key regulators for cell fate and embryonic development. To support this premise, it is important to understand whether or not systemic epigenetic changes coordinate with the progression of development. We have demonstrated that DNA methylation is programmed when neural stem cells differentiate (Zhou et al.,2011). Here, we analyzed the DNA methylation events that occur during early neural tube development. METHODS AND RESULTS: Using immunocytochemistry, we demonstrated that the DNA methylation marks - 5-methylcytosine (5-MeC), DNA methylation binding domain 1 (MBD1), and DNA methytransferases 1 (DNMT1) were highly coordinated in temporal and spatial patterns that paralleled the progress of embryonic development. The above ontogenic program of DNA methylation was, however, subjected to environmental modification. Alcohol exposure during fetal development, which is known to cause fetal alcohol spectrum disorder, altered the density and distribution of the DNA methylation marks. The alcohol exposure (88 mM) over 6 or 44 hours at gestation day 8 (GD-8) to GD-10 altered timely DNA methylation and retarded embryonic growth. We further demonstrated that the direct inhibiting of DNA methylation with 5-aza-cytidine (5-AZA) resulted in similar growth retardation. CONCLUSIONS: We identified a temporal and spatial cellular DNA methylation program after initial erasure, which parallels embryonic maturation. Alcohol delayed the cellular DNA methylation program and also retarded embryonic growth. Since direct inhibiting of DNA methylation resulted in similar retardation, alcohol thus can affect embryonic development through a epigenetic pathway. PMID- 21630421 TI - Associations between periconceptional alcohol consumption and craniosynostosis, omphalocele, and gastroschisis. AB - BACKGROUND: Alcohol consumption during pregnancy is known to be associated with certain birth defects, but the risk of other birth defects is less certain. The authors examined associations between maternal alcohol consumption during pregnancy and craniosynostosis, omphalocele, and gastroschisis among participants in the National Birth Defects Prevention Study, a large, multicenter case-control study. METHODS: A total of 6622 control infants and 1768 infants with birth defects delivered from 1997-2005 were included in the present analysis. Maternal alcohol consumption was assessed as any periconceptional consumption (1 month prepregnancy through the third pregnancy month), and by quantity-frequency, duration, and beverage type. Alcohol consumption throughout pregnancy was explored for craniosynostosis since the period of development may extend beyond the first trimester. Adjusted odds ratios (OR) and 95% confidence intervals (CI) were estimated using unconditional logistic regression analysis. OR were adjusted for age, race/ethnicity, and state of residence at time of infant's birth. Gastroschisis OR were also adjusted for periconceptional smoking. RESULTS: Periconceptional alcohol consumption and craniosynostosis showed little evidence of an association (OR = 0.92; CI: 0.78-1.08), but alcohol consumption in the second (OR = 0.65; CI: 0.47-0.92) and third trimesters (OR = 0.68; CI: 0.49-0.95) was inversely associated with craniosynostosis. Periconceptional alcohol consumption was associated with omphalocele (OR = 1.50; CI: 1.15-1.96) and gastroschisis (OR = 1.40; CI: 1.17-1.67). CONCLUSIONS: Results suggest that maternal periconceptional alcohol consumption is associated with omphalocele and gastroschisis, and second and third trimester alcohol consumption are inversely associated with craniosynostosis. PMID- 21630422 TI - Issue overview: epigenetic processes in development. PMID- 21630423 TI - Origin and development of the atrioventricular myocardial lineage: insight into the development of accessory pathways. AB - Defects originating from the atrioventricular canal region are part of a wide spectrum of congenital cardiovascular malformations that frequently affect newborns. These defects include partial or complete atrioventricular septal defects, atrioventricular valve defects, and arrhythmias, such as atrioventricular re-entry tachycardia, atrioventricular nodal block, and ventricular preexcitation. Insight into the cellular origin of the atrioventricular canal myocardium and the molecular mechanisms that control its development will aid in the understanding of the etiology of the atrioventricular defects. This review discusses current knowledge concerning the origin and fate of the atrioventricular canal myocardium, the molecular mechanisms that determine its specification and differentiation, and its role in the development of certain malformations such as those that underlie ventricular preexcitation. PMID- 21630424 TI - Modeling geographic risk of complex congenital heart defects in Eastern Wisconsin. AB - BACKGROUND: Geographic variation may be an indicator of risk factors for birth defects. This study models the geographic distribution of three complex congenital heart defects (CHDs) in eastern Wisconsin, and evaluates effects of demographic census variables linked to geographic location. METHODS: Cases of Hypoplastic Left Heart Syndrome (HLHS), Tetralogy of Fallot (TOF) and d Transposition of the Great Arteries (d-TGAs) born between1995 and 2004 were identified from three medical centers serving eastern Wisconsin. Case diagnoses were assigned by a pediatric cardiologist using echocardiographic records. Births by ZIP code were obtained from the State of Wisconsin. ZIP Code demographic variables were derived from 2000 census data. Numbers of cardiac defects by ZIP code were modeled using cluster analysis and Poisson generalized additive models (GAMs) for spatial coordinates including all and white only cases (excluding trisomies). GAM analyses were repeated adjusting for census variables. RESULTS: Four hundred forty-eight cases were ascertained. A significant south-to-north spatial gradient for HLHS, TOF, and combined CHDs, but not d-TGAs was identified. This gradient remained significant when census variables were included in the model for the full sample. In the analysis excluding non-white cases, findings were the same for TOF, combined CHDs, and d-TGAs. However, the geographic gradient for HLHS was not significant in the adjusted model. CONCLUSIONS: A south to-north gradient was apparent for two of three complex CHDs in eastern Wisconsin. For white cases, demographic variation seems to explain some of this spatial gradient in HLHS. Further studies are needed to confirm demographic and other risk factors underlying this geographic gradient. PMID- 21630425 TI - Evaluation of mailed pediatric buccal cytobrushes for use in a case-control study of birth defects. AB - BACKGROUND: Buccal cell collection is a convenient DNA collection method; however, little attention has been given to the quality of DNA obtained from pediatric populations. The purpose of this study was to determine the effect of a modified cytobrush collection method on the yield and quality of infant buccal DNA collected as part of a population-based case-control study of birth defects. METHODS Cytobrushes were collected from infants, mothers, and fathers using a standard collection method in 1997 to 2003 and a modified protocol that allows air-drying of the cytobrushes after collection from 2003 to the present. Yield and quality of DNA from 1057 cytobrushes was assessed by quantitative PCR and short tandem repeat (STR) genotyping, respectively. RESULTS Air-dried cytobrushes from infants had higher median DNA yields (1300 ng) and STR completion rates (99.5%) than standard collection method cytobrushes (60 ng and 59.5%, respectively). A subset of DNA aliquots was genotyped for six single nucleotide polymorphisms (SNPs). Aliquots from both collection methods that passed the quality protocol (DNA concentration >1 ng/MUl, and successful amplification of >=1 STR) had high genotype completion rates (99-100%). The median DNA yield following whole genome amplification was more than twofold higher for air-dried than standard collection specimens (p < 0.001). CONCLUSION Yield and quality of buccal DNA collected from infants are improved by using a method that incorporates air-drying; however, DNA collected by both methods is suitable for genotyping if stringent quality control procedures are instituted. These findings may be helpful for future epidemiologic studies of birth defects and other adverse pediatric outcomes. PMID- 21630426 TI - Effects of folic acid fortification on spina bifida prevalence in Brazil. AB - BACKGROUND: To assess spina bifida birth prevalence changes after folic acid fortification of wheat and maize flours began in Brazil in June 2004. METHODS: Cross-sectional study of Brazilian live births in 2004 and 2006. Spina bifida birth prevalence from the Live Births Information System (SINASC: Sistema de Informacoes sobre Nascidos Vivos) in a prefortified period was compared to a period fortified with folic acid in each state. Observed prevalence rates in 2004 were used to calculate the expected prevalence rates in 2006 under the null hypothesis that both were similar. The observed/expected (O/E) ratios were tested by two-tailed Z-test. To minimize ascertainment differences among states, the O/E ratio of each one of the 27 Brazilian states was adjusted for the number of births with the Mantel-Haenszel statistic. RESULTS The reduction in spina bifida birth prevalence in 2006 was 39% (O/E = 0.61; 95% confidence interval [CI], 0.55 0.67), and 40% (O/E = 0.60; 95% CI, 0.53-0.68), after adjusting for state birth number. This reduction was significant (p < 0.0001), and heterogeneous among states (chi(2) = 72.96; p < 0.0001). CONCLUSIONS: Using SINASC data, there was a significant reduction in spina bifida birth prevalence in Brazil, probably related to the folic acid food fortification program. PMID- 21630427 TI - Calcium-mediated repression of beta-catenin and its transcriptional signaling mediates neural crest cell death in an avian model of fetal alcohol syndrome. AB - Fetal alcohol syndrome (FAS) is a common birth defect in many societies. Affected individuals have neurodevelopmental disabilities and a distinctive craniofacial dysmorphology. These latter deficits originate during early development from the ethanol-mediated apoptotic depletion of cranial facial progenitors, a population known as the neural crest. We showed previously that this apoptosis is caused because acute ethanol exposure activates G-protein-dependent intracellular calcium within cranial neural crest progenitors, and this calcium transient initiates the cell death. The dysregulated signals that reside downstream of ethanol's calcium transient and effect neural crest death are unknown. Here we show that ethanol's repression of the transcriptional effector beta-catenin causes the neural crest losses. Clinically relevant ethanol concentrations (22-78 mM) rapidly deplete nuclear beta-catenin from neural crest progenitors, with accompanying losses of beta-catenin transcriptional activity and downstream genes that govern neural crest induction, expansion, and survival. Using forced expression studies, we show that beta-catenin loss of function (via dominant negative T cell transcription factor [TCF]) recapitulates ethanol's effects on neural crest apoptosis, whereas beta-catenin gain-of-function in ethanol's presence preserves neural crest survival. Blockade of ethanol's calcium transient using Bapta-AM normalizes beta-catenin activity and prevents the neural crest losses, whereas ionomycin treatment is sufficient to destabilize beta-catenin. We propose that ethanol's repression of beta-catenin causes the neural crest losses in this model of FAS. beta-Catenin is a novel target for ethanol's teratogenicity. beta-Catenin/Wnt signals participate in many developmental events and its rapid and persistent dysregulation by ethanol may explain why the latter is such a potent teratogen. PMID- 21630428 TI - Combination testing (Stage 2) of the Anti-IGF-1 receptor antibody IMC-A12 with rapamycin by the pediatric preclinical testing program. AB - BACKGROUND: IMC-A12, a fully human antibody that blocks ligand binding to the Type 1 insulin-like growth factor receptor, and rapamycin, a selective inhibitor of mTORC1 signaling, have both demonstrated significant antitumor activity against PPTP solid tumor models. Here we have evaluated antitumor activity of each agent individually and in combination against nine tumor models. PROCEDURES: IMC-A12 was administered twice weekly and rapamycin was administered daily for 5 days per week for a planned 4 weeks. The impact of combining IMC-A12 with rapamycin was evaluated using two measures: (1) the "therapeutic enhancement" measure, and (2) a linear regression model for time-to-event to formally evaluate for sub- and supra-additivity for the combination compared to the agents used alone. RESULTS: Two osteosarcomas, and one Ewing sarcoma of the nine xenografts tested showed therapeutic enhancement. The combination effect was most dramatic for EW-5 for which PD2 responses of short duration were observed for both single agents and a prolonged PR response was observed for the combination. Both OS-2 and OS-9 showed significantly longer times to progression with the combination compared to either of the single agents, although objective response criteria were not met. CONCLUSIONS: The combination of IMC-A12 with rapamycin was well tolerated, and induced tumor responses that were superior to either single agent alone in several models. These studies confirm reports using other antibodies that inhibit IGF-1 receptor-mediated signaling that indicate enhanced therapeutic effect for this combination, and extend the range of histotypes to encompass additional tumors expressing IGF-1R where this approach may be effective. PMID- 21630429 TI - Vascular endothelial growth factor and fibroblast growth factor 2 delivery from spinal cord bridges to enhance angiogenesis following injury. AB - The host response to spinal cord injury can lead to an ischemic environment that can induce cell death and limits cell transplantation approaches to promote spinal cord regeneration. Spinal cord bridges that provide a localized and sustained release of vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (FGF-2) were investigated for their ability to promote angiogenesis and nerve growth within the injury. Bridges were fabricated by fusion of poly(lactide-co-glycolide) microspheres using a gas foaming/particulate leaching technique, and proteins were incorporated by encapsulation into the microspheres and/or mixing with the microspheres before foaming. Compared to the mixing method, encapsulation reduced the losses during leaching and had a slower protein release, while VEGF was released more rapidly than FGF-2. In vivo implantation of bridges loaded with VEGF enhanced the levels of VEGF within the injury at 1 week, and bridges releasing VEGF and FGF-2 increased the infiltration of endothelial cells and the formation of blood vessel at 6 weeks postimplantation. Additionally, substantial neurofilament staining was observed within the bridge; however, no significant difference was observed between bridges with or without protein. Bridges releasing angiogenic factors may provide an approach to overcome an ischemic environment that limits regeneration and cell transplantation-based approaches. PMID- 21630430 TI - Adipose tissue-derived stem cells display a proangiogenic phenotype on 3D scaffolds. AB - Ischemic heart disease is the leading cause of death worldwide. Recent studies suggest that adipose tissue-derived stem cells (ASCs) can be used as a potential source for cardiovascular tissue engineering due to their ability to differentiate along the cardiovascular lineage and to adopt a proangiogenic phenotype. To understand better ASCs' biology, we used a novel 3D culture device. ASCs' and b.END-3 endothelial cell proliferation, migration, and vessel morphogenesis were significantly enhanced compared to 2D culturing techniques. ASCs were isolated from inguinal fat pads of 6-week-old GFP+/BLI+ mice. Early passage ASCs cells (P3-P4), PKH26-labeled murine b.END-3 cells or a co-culture of ASCs and b.END-3 cells were seeded at a density of 1 * 10(5) on three different surface configurations: (a) a 2D surface of tissue culture plastic, (b) Matrigel, and (c) a highly porous 3D scaffold fabricated from inert polystyrene. VEGF expression, cell proliferation, and tubulization, were assessed using optical microscopy, fluorescence microscopy, 3D confocal microscopy, and SEM imaging (n = 6). Increased VEGF levels were seen in conditioned media harvested from co cultures of ASCs and b.END-3 on either Matrigel or a 3D matrix. Fluorescence, confocal, SEM, bioluminescence revealed improved cell, proliferation, and tubule formation for cells seeded on the 3D polystyrene matrix. Collectively, these data demonstrate that co-culturing ASCs with endothelial cells in a 3D matrix environment enable us to generate prevascularized tissue-engineered constructs. This can potentially help us to surpass the tissue thickness limitations faced by the tissue engineering community today. PMID- 21630431 TI - Osteoinductivity of partially purified bovine, ostrich and emu bone morphogenetic proteins in vitro. AB - The aim of this study was to observe the osteogenic activity of native bone morphogenetic proteins (BMPs) obtained from different species including bovine, ostrich and emu sources in order to compare mammalian and avian BMPs. Rat mesenchymal progenitor marrow stromal cells and pre-osteoblastic C2C12 cell cultures, were exposed to the native BMPs and alkaline phosphatase (ALP) and creatine kinase (CK) levels were determined by assay. The results showed that the ALP activity in C2C12 cultures was elevated by bovine BMP by 2- to 10-fold (p < 0.05-0.001) from day 3 during 14 days. There were no significant differences in avian BMP related elevations of ALP activity except with ostrich BMPs at day 14 (p < 0.05). However, exposure of MSCs cultures to BMPs derived from bovine, ostrich or emu sources resulted in elevated ALP from day 3 (p < 0.05). Bovine BMP resulted in more ALP elevation than with either of the avian BMPs. All of BMPs elevated Creatine kinase (CK) activity from day 1 and climbed until peaking at day 7. Compared with control cultures, CK was elevated more with exposure to emu BMP and was more elevated with greater statistical significance than with bovine and ostrich BMP before day 5. These higher levels remained until day 14 (p < 0.05). The results of this study suggest that both bovine and avian BMPs are able to stimulate osteogenesis in mature osteoblasts in vitro. The strongest synergistic effect on osteogenesis was detected in cells stimulated with bovine BMP. Avian BMPs had lower effects on ALP and CK activity, emu BMP being more effective than ostrich BMP. PMID- 21630432 TI - Structural surface changes and inflammatory responses against alginate-based microcapsules after exposure to human peritoneal fluid. AB - Microencapsulation of cells is a promising approach to prevent rejection in the absence of immunosuppression. Clinical application, however, is hampered by insufficient insight in factors influencing biocompatibility of the capsules in humans. In the present study we exposed alginate-based capsules prepared of different types of alginate to human peritoneal fluid. Subsequently we studied the physicochemical changes of the capsule's surface by applying micro-Fourier Transform Infrared Spectroscopy. We did test alginate-beads and alginate-poly-L lysine capsules prepared of different types of alginate. In all tested capsule formulations we found adsorption of components from human peritoneal fluid and clear physicochemical changes of the surface. These changes were alginate dependent. The adsorption had no significant effects on the permselective properties of the capsule but we found a strong increase of TNFalpha production by human peripheral blood mononuclear cells when exposed to alginate-beads treated with human peritoneal fluid. This elevated responsiveness was not observed with alginate-PLL capsules. The results show that alginate-based capsule surfaces always undergo physicochemical changes of the surface when exposed to human peritoneal fluid. This adsorption may lead to enhancement of the inflammatory responses against the microcapsules. Our result implicate that biocompatibility measurements should not only been done with freshly prepared capsules but also with capsules that have been exposed to fluid from the implantation site in order to predict the in vivo responses. PMID- 21630433 TI - Inhibition of hydrogen embrittlement of Ni-Ti superelastic alloy in acid fluoride solution by hydrogen peroxide addition. AB - Inhibition of the hydrogen embrittlement of Ni-Ti superelastic alloy in an acidulated phosphate fluoride (APF) solution has been attempted by adding various amounts of H(2)O(2). In a 0.2% APF solution, hydrogen absorption is markedly inhibited by adding H(2)O(2), although corrosion is slightly enhanced by increasing the amount of added H(2)O(2). By adding a small amount of H(2)O(2) (0.001 M), in the early stage of immersion, hydrogen embrittlement is inhibited and corrosion is only slightly enhanced. Upon adding H(2)O(2), it appears that the dominant cathodic reactions change from hydrogen evolution to H(2)O(2) reduction reactions, or the surface conditions of the alloy are changed by H(2)O(2) with a high oxidation capability, thereby inhibiting hydrogen absorption. The present study clearly indicates that infinitesimal addition of H(2)O(2) into acid fluoride solutions is effective for the inhibition of the hydrogen embrittlement of the alloy. PMID- 21630434 TI - Chondrogenic differentiation of neonatal human dermal fibroblasts encapsulated in alginate beads with hydrostatic compression under hypoxic conditions in the presence of bone morphogenetic protein-2. AB - In the present work, neonatal human dermal fibroblasts (nHDFs) were evaluated as a potential cell source for intervertebral disc repair. Chondrogenic differentiation of nHDFs was studied in the presence or absence of hydrostatic compression under normal and hypoxic conditions. In addition, the potentially synergistic effects of mechanical stimulation and bone morphogenetic protein (BMP)-2 on the chondrogenic differentiation of nHDFs were assessed. Mechanical stimulation was applied to the cells encapsulated in alginate beads using a custom-built bioreactor system for either a 1- or 3-week period at a frequency of 1 Hz for 4 h/day. In general, after 21 days of culture, high cell viability was observed for all the groups, with the exception of the groups exposed to intermittent mechanical stimulation for 3 weeks. Long-term intermittent application of mechanical stimulation under low O(2) conditions resulted in elevated collagen biosynthesis rate from day 0. Inclusion of BMP-2 for this group improved the chondrogenic differentiation of nHDFs, as indicated by elevated aggrecan gene expression and an increased collagen production rate compared to the day 0 group. Thus, the combination of hypoxia, BMP-2 supplementation, and long-term intermittent application of dynamic hydrostatic pressure was found to be a potent promoter of the chondrogenic differentiation of nHDFs. PMID- 21630435 TI - Reduction of protein adsorption and macrophage and astrocyte adhesion on ventricular catheters by polyethylene glycol and N-acetyl-L-cysteine. AB - Cellular obstruction of poly(dimethyl)siloxane (PDMS) catheters is one of the most prevalent causes of shunt failure in the treatment of hydrocephalus. By modifying PDMS using short- and long-chain mono-functional polyethylene glycol (PEG604 and PEG5K, respectively) and N-acetyl-L-cysteine via adsorption and covalent binding (NAC and NAC/EDC/NHS, respectively), we increased surface wettability. We hypothesized that these surface modifications would inhibit protein adsorption and decrease host macrophage and astrocyte adhesion. Tested in a bioreactor set to mimic physiological flow, all modified surfaces significantly decreased albumin adsorption compared with PDMS (p < 0.05) except for PEG604 modified PDMS (p = 0.14). All four modification strategies significantly reduced (p < 0.01) fibronectin adsorption. PEG604, PEG5K, NAC, and NAC/EDC/NHS reduced the average level of macrophage adhesion by 53%, 63%, 40%, and 58% (p <.0.05 except when comparing PDMS with NAC) and astrocyte adhesion by 47%, 83%, 91%, and 72% (p < 0.05 except when comparing PDMS with PEG604), respectively. Combined with saline soak results which suggest that the surface wettability is stable over 30 days for each modification, our results are consistent with the hypothesis that these modifications decrease cell adhesion on catheters in vitro for the treatment of hydrocephalus. PMID- 21630436 TI - Three-dimensional honeycomb-patterned chitosan/poly(L-lactic acid) scaffolds with improved mechanical and cell compatibility. AB - Micro-size patterned surfaces trigger specific biological responses such as the promotion of cell growth, cell migration, cell differentiation, and ECM production. The aim of this work was to elaborate three-dimensional scaffolds with honeycomb patterned surfaces and large open pores, and to study the influence of surface patterning on cell behavior. In this study, we used water droplets as porogen material to prepare a novel type of chitosan sponge with large open pores on its surface. The sponges obtained were then immersed into 6 wt % Poly(L-lactic acid) chloroform solution to obtain honeycomb patterned composite porous scaffolds. The morphology and mechanical properties were characterized with SEM and compression testing. The fibroblast behaviors in scaffolds were analyzed with SEM, VG, PAS, live-dead staining, and flow cytometer. Results showed that these composite scaffolds possessed better mechanical properties and hierarchical porous structure than pure chitosan sponges. Cell culture revealed that the honeycomb patterned surface had positive influences on fibroblast behaviors, wherein the cell adhesion, proliferation, ECM secretion and viability were improved dramatically. Such a hierarchical composite scaffold would be a suitable candidate for tissue engineering purposes. PMID- 21630437 TI - A follow-up case-control association study of tractable (druggable) genes in recurrent major depression. AB - The High-Throughput Disease-specific target Identification Program (HiTDIP) aimed to study case-control association samples for 18 common diseases. Here we present the results of a follow-up case-control association study of HiTDIP in major depressive disorder (MDD). The HiTDIP in MDD was conducted in a sample of 974 cases of recurrent MDD of white German origin collected at the Max-Planck Institute (MP-GSK) and 968 ethnically matched controls screened for lifetime absence of depression. Six genes were identified as of interest for a follow-up, based on the strength of the association and based on the interest as potential candidate target for developing new treatment for depression: Solute Carrier Family 4 Member 10 (SLC4A10), Dipeptidyl Peptidase IV (DPP4), Dopamine Receptor D3 (DRD3), Zinc Finger Protein 80 (ZNF80), Nitric Oxide Synthase 2A (NOS2A) and Peroxisome Proliferator-Activated Receptor-Gamma, Coactivator 1, Alpha (PPARGC1A). Within the current study, we attempted to follow-up these findings in a sample from the UK, the Depression Case Control (DeCC) sample consisting of 1,196 cases and 842 screened controls, phenotyped using exactly the same methods as the MP-GSK sample. Performing Cochran-Mantel-Haenzel statistics to test for genotypic and/or allelic differences between the DeCC and MP-GSK samples, we found no significant differences, thus being able to combine the two samples for association testing. In the combined sample of 2,170 MDD cases and 1,810 controls, there were positive findings in the Nitric Oxide Synthase 2A (NOS2A) gene both using single SNP analysis and haplotype analysis. PMID- 21630439 TI - Effects on human health, functionalities, and occurrence of phenolic compounds. PMID- 21630440 TI - Graphene-based materials: synthesis, characterization, properties, and applications. AB - Graphene, a two-dimensional, single-layer sheet of sp(2) hybridized carbon atoms, has attracted tremendous attention and research interest, owing to its exceptional physical properties, such as high electronic conductivity, good thermal stability, and excellent mechanical strength. Other forms of graphene related materials, including graphene oxide, reduced graphene oxide, and exfoliated graphite, have been reliably produced in large scale. The promising properties together with the ease of processibility and functionalization make graphene-based materials ideal candidates for incorporation into a variety of functional materials. Importantly, graphene and its derivatives have been explored in a wide range of applications, such as electronic and photonic devices, clean energy, and sensors. In this review, after a general introduction to graphene and its derivatives, the synthesis, characterization, properties, and applications of graphene-based materials are discussed. PMID- 21630441 TI - Facile synthesis of gold nanorice enclosed by high-index facets and its application for CO oxidation. AB - A facile method for generating Au nanorice enclosed by high-index facets in high purity. The nanorice shows much higher catalytic activity for CO oxidation than multiply twinned particles of Au enclosed by {111} facets at temperatures below 300 degrees C. PMID- 21630442 TI - One-step high-yield aqueous synthesis of size-tunable multispiked gold nanoparticles. AB - Multispiked gold nanoparticles are required in large quantities for many fundamental studies and applications like (bio)sensing, but their preparation in high yield by the bottom-up chemical synthetic method is challenging. A water based, non-'seed-mediated', straightforward method for the synthesis of gold nanoparticles with well-developed surface spikes is reported here. The yield of multispiked gold particles is very high (>90%). The method allows the tuning of the number and size of the spikes and the overall size of the particles, and hence the localized surface plasmon resonances of the particles over the broad spectral range in the visible and near-infrared. A mechanism for the evolution of twinned, sharp-tipped surface protrusions has been proposed based on systematic spectrophotometric and transmission electron microscopic studies, which were employed to elucidate the morphological features, structure, chemical composition, and optical properties of the multispiked gold nanoparticles. PMID- 21630443 TI - Growth inhibition of Staphylococcus aureus by mixed monolayer gold nanoparticles. PMID- 21630444 TI - Cisplatin-loaded porous Si microparticles capped by electroless deposition of platinum. AB - The loading and release of the anti-cancer drug platinum cis-dichlorodiamine (cisplatin) from mesoporous silicon (pSi) microparticles is studied. The pSi microparticles are modified with 1-dodecene or with 1,12-undecylenic acid by hydrosilylation, and each modified pSi material acts as a reducing agent, forming a deposit of Pt on its surface that nucleates further deposition, capping the mesoporous structure and trapping free (unreduced) cisplatin within. Slow oxidation and hydrolytic dissolution of the Si/SiO(2) matrix in buffer solution or in culture medium leads to the release of drugs from the microparticles. The drug-loaded particles show significantly greater toxicity toward human ovarian cancer cells (in vitro), relative to an equivalent quantity of free cisplatin. This result is consistent with the mechanism of drug release, which generates locally high concentrations of the drug in the vicinity of the degrading particles. Control assays with pSi particles loaded in a similar manner with the therapeutically inactive trans isomer of the platinum drug, and with pSi particles containing no drug, result in low cellular toxicity. A hydrophobic pro drug, cis,trans,cis-[Pt(NH(3))(2)(O(2)C(CH(2))(8)CH(3))(2)Cl(2)], is loaded into the pSi films from chloroform without concomitant reduction of the pSi carrier. PMID- 21630445 TI - Anisotropic adhesion properties of triangular-tip-shaped micropillars. AB - Directional dry adhesive microstructures consisting of high-density triangular tip-shaped micropillars are described. The wide-tip structures allow for unique directional shear adhesion properties with respect to the peeling direction, along with relatively high normal adhesion. PMID- 21630447 TI - Self-assembled large Au nanoparticle arrays with regular hot spots for SERS. AB - The cost-effective self-assembly of 80 nm Au nanoparticles (NPs) into large domain, hexagonally close-packed arrays for high-sensitivity and high-fidelity surface-enhanced Raman spectroscopy (SERS) is demonstrated. These arrays exhibit specific optical resonances due to strong interparticle coupling, which are well reproduced by finite-difference time-domain (FDTD) simulations. The gaps between NPs form a regular lattice of hot spots that enable a large amplification of both photoluminescence and Raman signals. At smaller wavelengths the hot spots are extended away from the minimum-gap positions, which allows SERS of larger analytes that do not fit into small gaps. Using CdSe quantum dots (QDs) a 3-5 times larger photoluminescence enhancement than previously reported is experimentally demonstrated and an unambiguous estimate of the electromagnetic SERS enhancement factor of ~10(4) is obtained by direct scanning electron microscopy imaging of QDs responsible for the Raman signal. Much stronger enhancement of ~10(8) is obtained at larger wavelengths for benzenethiol molecules penetrating the NP gaps. PMID- 21630446 TI - Addressing brain tumors with targeted gold nanoparticles: a new gold standard for hydrophobic drug delivery? AB - EGF-modified Au NP-Pc 4 conjugates showed 10-fold improved selectivity to the brain tumor compared to untargeted conjugates. The hydrophobic photodynamic therapy drug Pc 4 can be delivered efficiently into glioma brain tumors by EGF peptide-targeted Au NPs. Compared to the untargeted conjugates, EGF-Au NP-Pc 4 conjugates showed 10-fold improved selectivity to the brain tumor. This delivery system holds promise for future delivery of a wider range of hydrophobic therapeutic drugs for the treatment of hard-to-reach cancers. PMID- 21630448 TI - Proteomic analysis reveals the strategies of Bacillus thuringiensis YBT-1520 for survival under long-term heat stress. AB - Bacillus thuringiensis (Bt) has been widely used for 50 years as a safe biopesticide for controlling agricultural and sanitary insect pests because of its insecticidal crystal proteins. In this study a proteomic approach was used to investigate the responses and survival strategies of Bt YBT-1520 under a long term heat stress condition (42 degrees C). Heat stress mainly influenced the characteristics of YBT-1520 on four aspects: (i) the abilities to synthesise insecticidal crystal proteins and other potential pathogenic factors were almost lost, (ii) cell adhesion and motility were also lost, (iii) cell did not sporulate, (iv) cell kept accumulating poly(3-hydroxybutyrate) (PHB). Proteomic analyses to the physiological changes of the strain revealed three strategies of YBT-1520 for survival under long-term heat stress. The first strategy is to up regulate enzymes (BDH1, GuaB and PepA) for long-term heat stress tolerance. The second one is to down-regulate metabolic enzymes to reduce metabolic burden. The third strategy is to increase the synthesis and accumulation of PHB. Under heat stress condition, the bacterium adjusted its metabolism by up-/down-regulation and continuous accumulation of PHB. These strategies would help cells to gain more tolerance to heat stress. PMID- 21630449 TI - Four-dimensional visualisation and analysis of protein-protein interaction networks. AB - Protein-protein interaction networks are typically built with interactions collated from many experiments. These networks are thus composite and show all interactions that are currently known to occur in a cell. However, these representations are static and ignore the constant changes in protein-protein interactions. Here we present software for the generation and analysis of dynamic, four-dimensional (4-D) protein interaction networks. In this, time course-derived abundance data are mapped onto three-dimensional networks to generate network movies. These networks can be navigated, manipulated and queried in real time. Two types of dynamic networks can be generated: a 4-D network that maps expression data onto protein nodes and one that employs 'real-time rendering' by which protein nodes and their interactions appear and disappear in association with temporal changes in expression data. We illustrate the utility of this software by the analysis of singlish interface date hub interactions during the yeast cell cycle. In this, we show that proteins MLC1 and YPT52 show strict temporal control of when their interaction partners are expressed. Since these proteins have one and two interaction interfaces, respectively, it suggests that temporal control of gene expression may be used to limit competition at the interaction interfaces of some hub proteins. The software and movies of the 4-D networks are available at http://www.systemsbiology.org.au/downloads_geomi.html. PMID- 21630450 TI - A cost-benefit analysis of multidimensional fractionation of affinity purification-mass spectrometry samples. AB - Affinity purification coupled to mass spectrometry (AP-MS) is gaining widespread use for the identification of protein-protein interactions. It is unclear, however, whether typical AP sample complexity is limiting for the identification of all protein components using standard one-dimensional LC-MS/MS. Multidimensional sample separation is useful for reducing sample complexity prior to MS analysis and increases peptide and protein coverage of complex samples. Here, we monitored the effects of upstream protein or peptide separation techniques on typical mammalian AP-MS samples, generated by FLAG affinity purification of four baits with different biological functions and/or subcellular distribution. As a first separation step, we employed SDS-PAGE, strong cation exchange LC, or reversed-phase LC at basic pH. We also analyzed the benefits of using an instrument with a faster scan rate, the new TripleTOF 5600 mass spectrometer. While all multidimensional approaches yielded a clear increase in spectral counts, the increase in unique peptides and additional protein identification was modest and came at the cost of increased instrument and handling time. The use of a high duty-cycle instrument achieved similar benefits without these drawbacks. An increase in spectral counts is beneficial when data analysis methods relying on spectral counts, including Significance Analysis of INTeractome (SAINT), are used. PMID- 21630451 TI - Constructing the metabolic and regulatory pathways in germinating rice seeds through proteomic approach. AB - Construction of metabolic and regulatory pathways from proteomic data can contextualize the large-scale data within the overall physiological scheme of an organism. It is an efficient way to predict metabolic phenotype or regulatory style. We did protein profiling in the germinating rice seeds through 1-DE via LC MS/MS proteomic shotgun strategy. In total, 673 proteins were identified, and could be sorted into 14 functional groups. The largest group was metabolism related. The metabolic proteins were integrated into different metabolic pathways to show the style of reserves mobilization and precursor preparation during the germination. Analysis of the regulatory proteins indicated that regulation of redox homeostasis and gene expression also play important roles for the rice seed germination. Although transcription is unnecessary for the germination, it could ensure the rapidity and uniformity of germination. On the contrary, translation with the stored mRNA is required for the germination. This study will help us to further understand the metabolic style, regulation of redox homeostasis, and gene expression during rice seed germination. PMID- 21630452 TI - Using 2-DE for the differentiation of local chicken breeds. AB - The aim of this study was to apply a proteomic approach for the characterisation of local chicken breeds. The experiment involved a total of 29 males of three local Italian chicken breeds: Pepoi, Padovana and Ermellinata di Rovigo. Sarcoplasmic protein fractions of breast muscle were analysed by 2-DE. Image analysis followed by statistical analysis enabled to differentiate groups of individuals based on the similarities of protein expression. Individuals were distinguished into clusters and groups, corresponding to the breed of origin. Distances among individuals, calculated using data on spot volumes, were used to draw a neighbour-joining tree, showing clear individual and breed grouping. The most relevant spots regarding breed differentiation were detected; 11 were identified by MS revealing preliminary evidences on the mechanisms of the breed differentiation process. The results evidenced the ability of proteomic analyses for the characterisation of chicken breeds. PMID- 21630453 TI - Rapid isolation and characterization of bacterial lipopeptides using liquid chromatography and mass spectrometry analysis. AB - The structural analysis of post-translational modifications (PTMs) of lipoproteins is difficult due to the hydrophobic properties of their fatty acid moieties. At the present time, the relative positions of fatty acid components on the N-acyl-S-diacylglycerylcysteine core structure has not been specifically identified in any natural or bacterial expressed recombinant lipoproteins. In this study, we describe a rapid solid-phase extraction using acetonitrile and isopropanol method that can be performed manually to isolate large amounts of relatively pure lipopeptides generated by the limited tryptic-digestion of recombinant lipoproteins. Using these lipopeptides and LC/MS mass spectra analysis, two groups of N-terminal lipidated (diacyl or triacyl) molecules that differ by one fatty acid unit were successfully identified. This LC/MS method also provided the separation of lipopeptides differing by 14 Da for the on-line MS identification. Multiple-stage fragmentation analyses of the di- and triacyl lipopeptides using both the positive and negative ion modes enabled to identify the putative structure of the N-acyl-S-diacylglycerylcysteine containing an amide bond to palmitic acid at the N-terminal cysteine, a palmitic acid at sn1 position, and an unsaturated fatty acid of either hexadecenoic acid, cyclopropaneoctanoic acid, oleic acid and nonadecenoic acid at sn2 position of diacylglycerol residue through ester bonding. For diacyl lipoprotein, the saturated palmitoyl fatty acid group is absent at sn1 position of glycerol derived lipid residue of lipopeptide. PMID- 21630454 TI - Quantitative peptidomic analysis by a newly developed one-step direct transfer technology without depletion of major blood proteins: its potential utility for monitoring of pathophysiological status in pregnancy-induced hypertension. AB - We have recently developed a new target plate (BLOTCHIP(r)) for MALDI-MS. An advantage of this procedure is that it does not require the lowering of protein concentrations in test samples prior to analysis. Accordingly, this new technology enables the detection of peptides present in blood samples, including those that would otherwise be adsorbed to abundant blood proteins and would thus escape detection. Using this technology, we analyzed the peripheral blood of patients with pregnancy-induced hypertension (PIH; the most common serious complication of pregnancy) to test a potential utility of the technology for monitoring of the pathophysiological status. In the present study, we found 23 characteristic peptides for PIH in the blood serum of pregnant women. Offline LC MALDI MS/MS identified 7 of the 23 peptides as fragments derived from kininogen-1 (three peptides), fibrinogen-alpha, complement component C4-A/B, alpha-2-HS glycoprotein and inter-alpha-trypsin inhibitor heavy chain H4. 2-D scatter plots with combinations of the peptides found in the present study can be grouped for pregnant women with/without PIH, which would be satisfactory reflected for their status. Additionally, the levels of most of these peptides found were significantly decreased by albumin/IgG depletion prior to BLOTCHIP(r) analysis in accordance with conventional proteomics procedures. These results indicated that BLOTCHIP(r) analysis can be applied for discovery study of PIH biomarker candidates. PMID- 21630455 TI - Proteomic analysis of papaya (Carica papaya L.) displaying typical sticky disease symptoms. AB - Papaya (Carica papaya L.) hosts the only described laticifer-infecting virus (Papaya meleira virus, PMeV), which is the causal agent of papaya sticky disease. To understand the systemic effects of PMeV in papaya, we conducted a comprehensive proteomic analysis of leaf samples from healthy and diseased plants grown under field conditions. First, a reference 2-DE map was established for proteins from healthy samples. A total of 486 reproducible spots were identified, and MALDI-TOF-MS/MS data identified 275 proteins accounting for 159 distinct proteins from 231 spots that were annotated. Second, the differential expression of proteins from healthy and diseased leaves was determined through parallel experiments, using 2-DE and DIGE followed by MALDI-TOF-MS/MS and LC-IonTrap MS/MS, respectively. Conventional 2-DE analysis revealed 75 differentially expressed proteins. Of those, 48 proteins were identified, with 26 being upregulated (U) and 22 downregulated (D). In general, metabolism-related proteins were downregulated, and stress-responsive proteins were upregulated. This expression pattern was corroborated by the results of the DIGE analysis, which identified 79 differentially expressed proteins, with 23 identified (17 U and 6 D). Calreticulin and the proteasome subunits 20S and RPT5a were shown to be upregulated during infection by both 2-DE and DIGE analyses. These data may help shed light on plant responses against stresses and viral infections. PMID- 21630456 TI - Nanoprobe-based immobilized metal affinity chromatography for sensitive and complementary enrichment of multiply phosphorylated peptides. AB - Magnetic nanoparticles (MNP, <100 nm) have rapidly evolved as sensitive affinity probes for phosphopeptide enrichment. By taking advantage of the easy magnetic separation and flexible surface modification of the MNP, we developed a surface blocked, nanoprobe-based immobilized metal ion affinity chromatography (NB-IMAC) method for the enhanced purification of multiply phosphorylated peptides. The NB IMAC method allowed rapid and specific one-step enrichment by blocking the surface of titanium (IV) ion-charged nitrilotriacetic acid-conjugated MNP (Ti4 NTA-PEG@MNP) with low molecular weight polyethylene glycol. The MNP demonstrated highly sensitive and unbiased extraction of both mono- and multiply phosphorylated peptides from diluted beta-casein (2 * 10-10 M). Without chemical derivation or fractionation, 1283 phosphopeptides were identified from 400 MUg of Raji B cells with 80% purification specificity. We also showed the first systematic comparison on the particle size effect between nano-sclae IMAC and micro-scale IMAC. Inductively coupled plasma-mass spectrometry (ICP-MS) analysis revealed that MNP had a 4.6-fold higher capacity for metal ions per unit weight than did the magnetic micro-sized particle (MMP, 2-10 MUm), resulting in the identification of more phosphopeptides as well as a higher percentage of multiply phosphorylated peptides (31%) at the proteome scale. Furthermore, NB-IMAC complements chromatography-based IMAC and TiO2 methods because <13% of mono- and 12% of multiply phosphorylated peptide identifications overlapped among the 2700 phosphopeptides identified by the three methods. Notably, the number of multiply phosphorylated peptides was enriched twofold and threefold by NB-IMAC relative to micro-scale IMAC and TiO2, respectively. NB-IMAC is an innovative material for increasing the identification coverage in phosphoproteomics. PMID- 21630457 TI - ProteoConnections: a bioinformatics platform to facilitate proteome and phosphoproteome analyses. AB - Novel and improved computational tools are required to transform large-scale proteomics data into valuable information of biological relevance. To this end, we developed ProteoConnections, a bioinformatics platform tailored to address the pressing needs of proteomics analyses. The primary focus of this platform is to organize peptide and protein identifications, evaluate the quality of the acquired data set, profile abundance changes, and accelerate data interpretation. Peptide and protein identifications are stored into a relational database to facilitate data mining and to evaluate the quality of data sets using graphical reports. We integrated databases of known PTMs and other bioinformatics tools to facilitate the analysis of phosphoproteomics data sets and to provide insights for subsequent biological validation experiments. Phosphorylation sites are also annotated according to kinase consensus motifs, contextual environment, protein domains, binding motifs, and evolutionary conservation across different species. The practical application of ProteoConnections is further demonstrated for the analysis of the phosphoproteomics data sets from rat intestinal IEC-6 cells where we identified 9615 phosphorylation sites on 2108 phosphoproteins. Combined proteomics and bioinformatics analyses revealed valuable biological insights on the regulation of phosphoprotein functions via the introduction of new binding sites on scaffold proteins or the modulation of protein-protein, protein-DNA, or protein-RNA interactions. Quantitative proteomics data can be integrated into ProteoConnections to determine the changes in protein phosphorylation under different cell stimulation conditions or kinase inhibitors, as demonstrated here for the MEK inhibitor PD184352. PMID- 21630458 TI - An expanded protein-protein interaction network in Bacillus subtilis reveals a group of hubs: Exploration by an integrative approach. AB - We have generated a protein-protein interaction network in Bacillus subtilis focused on several essential cellular processes such as cell division, cell responses to various stresses, the bacterial cytoskeleton, DNA replication and chromosome maintenance by careful application of the yeast two-hybrid approach. This network, composed of 793 interactions linking 287 proteins with an average connectivity of five interactions per protein, represents a valuable resource for future functional analyses. A striking feature of the network is a group of highly connected hubs (GoH) linking many different cellular processes. Most of the proteins of the GoH have unknown functions and are associated to the membrane. By the integration of available knowledge, in particular of transcriptome data sets, the GoH was decomposed into subgroups of party hubs corresponding to protein complexes or regulatory pathways expressed under different conditions. At a global level, the GoH might function as a very robust group of date hubs having partially redundant functions to integrate information from the different cellular pathways. Our analyses also provide a rational way to study the highly redundant functions of the GoH by a genetic approach. PMID- 21630459 TI - Proteomic characterization of the human sperm nucleus. AB - Generating a catalogue of sperm nuclear proteins is an important first step towards the clarification of the function of the paternal chromatin transmitted to the oocyte upon fertilization. With this goal, sperm nuclei were obtained through CTAB treatment and isolated to over 99.9% purity without any tail fragments, acrosome or mitochondria as assessed by optical microscopy and transmission electron microscopy. The nuclear proteins were extracted and separated in 2-D and 1-D gels and the 2-D spots and 1-D bands were excised and analysed to identify the proteins through LC-MS/MS. With this approach, 403 different proteins have been identified from the isolated sperm nuclei. The most abundant family of proteins identified are the histones, for which several novel members had not been reported previously as present in the spermatogenic cell line or in the human mature spermatozoa. More than half (52.6%) of the proteins had not been detected in the previous human whole sperm cell proteome reports. Of relevance, several chromatin-related proteins, such as zinc fingers and transcription factors, so far not known to be associated with the sperm chromatin, have also been detected. This provides additional information about the nuclear proteins that are potentially relevant for epigenetic marking, proper fertilization and embryo development. PMID- 21630460 TI - Quantitative chemical proteomics in small-scale culture of phorbol ester stimulated basal breast cancer cells. AB - Basal-like breast cancers are commonly negative for expression of estrogen and progesterone receptors and HER-2 (triple-negative breast cancer), which makes this subtype of breast cancers more aggressive and less responsive to standard treatment. We have applied a small-scale chemical proteomics method using bisindolylmaleimide (Bis) class of protein kinase C inhibitors to study the Bis binding proteome in a cell culture model of basal breast carcinoma (MDA-MB-231). Using MS, we identified 174 proteins captured by the Bis-probe in phorbol ester (PMA) stimulated cells. Gene ontology analysis broadly categorised these proteins as ATP binding (42%), GTP binding (6%) and having nucleoside-triphosphatase activity (21%). Of the 64 enzymes captured by the Bis-probe, the majority had either ATP and/or nucleotide binding functions. Two previously unreported Bis binding protein kinases, serine/arginine-rich protein-specific kinase 1 (SRPK1) and interferon-induced RNA-dependent protein kinase (PKR) were observed. We then incorporated SILAC for quantitation to examine the proteins that were differentially captured by the Bis-probe following 30 and 60 min PMA stimulation. This provided novel evidence for PMA regulation of the enzymes glyceraldehyde-3 phosphate dehydrogenase, nucleolar RNA helicase 2 and Heterogeneous nuclear ribonucleoprotein M. PMID- 21630461 TI - The cell surface proteome of Staphylococcus aureus. AB - The Gram-positive bacterium Staphylococcus aureus is a wide spread opportunistic pathogen that can cause a range of life-threatening diseases. To obtain a better understanding of the global mechanisms for pathogenesis and to identify novel targets for therapeutic interventions, the S. aureus proteome has been recently 'dissected' in several studies. Proteins that are exposed on the cell surface - collectively referred to as the 'surfacome' - have received particular attention, because they can directly interact with extracellular molecules, including drugs and antibodies. Accordingly, these proteins represent interesting candidate targets for active or passive immunization against S. aureus. Here, we review the proteomics strategies used, and we compare the results that were so far obtained. Since the surfacome is part of the cell wall proteome, we first present an overview of general properties of the S. aureus cell envelope, cell wall associated proteins and mechanisms for protein attachment to the cell wall. Then we zoom in on the surfacome, and discuss the pro's and con's of the specific strategies that have been applied for surfacome profiling. The insights thus obtained may serve as leads for future studies on the S. aureus surfacome and possible applications. PMID- 21630462 TI - Proteomic analysis of microvesicles derived from human colorectal cancer ascites. AB - The presence of malignant ascites in the peritoneal cavity is a poor prognostic indicator of low survival rate. Various cancer cells, including those of colorectal cancer (CRC), release microvesicles (exosomes) into surrounding tissues and peripheral circulation including malignant ascites. Although recent progress has revealed that microvesicles play multiple roles in tumor progression, the protein composition and the pathological function of malignant ascites-derived microvesicles are still unknown. Here, we report the first global proteomic analyses of highly purified microvesicles derived from human CRC ascites. With 1-D SDS-PAGE and nano-LC-MS/MS analyses, we identified a total of 846 microvesicular proteins from ascites of three CRC patients with high confidence; 384 proteins were identified in at least two patients. We identified proteins that might function in tumor progression via disruption of epithelial polarity, migration, invasion, tumor growth, immune modulation, and angiogenesis. Furthermore, we identified several potential diagnostic markers of CRC including colon-specific surface antigens. Our proteomic analyses will help to elucidate diverse functions of microvesicles in cancer progression and will aid in the development of novel diagnostic tools for CRC. PMID- 21630463 TI - Proteomics analysis of Bifidobacterium longum NCC2705 growing on glucose, fructose, mannose, xylose, ribose, and galactose. AB - To investigate the molecular mechanisms underlying carbohydrate uptake and connected metabolic pathways of Bifidobacterium longum NCC2705, the proteomic profiles of bacteria grown on different carbon sources including glucose, fructose, mannose, xylose, ribose, and galactose were analyzed. Our results show that all sugars tested were catabolized via the bifid shunt. Sixty-eight proteins that exhibited changes in abundance of threefold or greater were identified by MS. A striking observation was the differential expression of proteins related to the pyruvate metabolism. Further analysis of acetic acid and lactic acid in the culture supernatants by HPLC at the end of fermentation showed that more lactic acid was produced during growth on fructose, ribose, xylose, galactose and more acetic acid was produced during the fermentation of glucose and mannose. Growth experiments revealed that B. longum NCC2705 preferentially used fructose, ribose, xylose, and galactose with higher growth rates over glucose and mannose. Furthermore, five proteins (GroEL, Eno, Tal, Pgm, and BL0033) exhibited clear phosphorylation modifications at serine and/or tyrosine residues. BL0033, a component of an ATP-binding cassette (ABC) transporter, was significantly more abundant in bacteria grown on fructose and, to a lesser extent, ribose and xylose. RT-PCR analysis revealed that all genes of the ABC transporter are induced in the presence of these sugars suggesting that BL0033, BL0034, BL0035, and BL0036 constitute an ABC transporter with fructose as preferred substrate. PMID- 21630467 TI - Editorial: In vivo protein folding--at the crossroad between basic research and biotechnology. PMID- 21630470 TI - Chemistry and biological activity of platinum amidine complexes. AB - Platinum amidine complexes represent a new class of potential antitumor drugs that contain the imino moiety HN=C(sp(2)) bonded to the platinum center. They can be related to the iminoether derivatives, which were recently shown to be the first Pt(II) compounds with a trans configuration endowed with anticancer activity. The chemical and biological properties of platinum amidine complexes, and more generally of platinum imino derivatives, can be rationally modified through suitable synthetic procedures with the aim of improving their cytotoxicity and antitumor activity. The addition of protic nucleophiles to nitriles coordinated to platinum in various oxidation states can offer a wide variety of complexes with chemical, structural, and physical properties specifically tuned for a more efficacious biological response. PMID- 21630471 TI - Barfly: sculpting membranes at the Drosophila neuromuscular junction. AB - The ability of a cell to change the shape of its membranes is intrinsic to many cellular functions. Proteins that can alter or recognize curved membrane structures and those that can act to recruit other proteins which stabilize the membrane curvature are likely to be essential in cell functions. The BAR (Bin, amphiphysin, RVS167 homology) domain is a protein domain that can either induce lipidic membranes to curve or can sense curved membranes. BAR domains are found in several proteins at neuronal synapses. We will review BAR domain structure and the role that BAR domain containing proteins play in regulating the morphology and function of the Drosophila neuromuscular junction. In flies the BAR domain containing proteins, endophilin and syndapin affect synaptic vesicle endocytosis, whereas CIP4, dRich, nervous wreck and syndapin affect synaptic morphology. We will review the growing evidence implicating mutations in BAR domain containing proteins being the cause of human pathologies. PMID- 21630472 TI - Extrinsic factors as multifunctional regulators of retinal ganglion cell morphogenesis. AB - Neurons acquire a unique cell-type dependent morphology during development that is critical for their function in a neural circuit. The process involves a neuron sending out an axon that grows in a directed fashion to its target, and the elaboration of multiple, branched dendrites. The ultimate morphology of the neuron is sculpted by factors in the environment that act directly or indirectly to influence the behavior of the growing axon and dendrites. The output neuron of the retina, the retinal ganglion cell (RGC), has served as a useful model for the identification of molecular signals that control neuronal morphogenesis, because the entire development of the neuron, from the initiation of neurites to the establishment of synapses, is accessible for experimental manipulation and visualization. In this review we discuss data which argue that the visual system uses a limited number of signals to control RGC morphogenesis, with single molecules being reused multiple times to control distinct events in axon and dendrite outgrowth. PMID- 21630474 TI - Neurochemical and structural organization of the principal nucleus of the inferior olive in the human. AB - The inferior olive (IO) is the sole source of the climbing fibers that innervate the Purkinje cells of the cerebellar cortex. The IO comprises several subdivisions, the dorsal accessory olive (DAO), medial accessory olive (MAO), and principal nuclei of the IO (IOpr); the relative sizes of these subnuclei vary among species. In human, there is an expansion of the cerebellar hemispheres and a corresponding expansion of the IOpr. We have examined the structural and neurochemical organization of the human IOpr, using sections stained with cresyl violet (CV) or immunostained for the calcium-binding proteins calbindin (CB), calretinin (CR), and parvalbumin (PV), the synthetic enzyme for nitric oxide (nNOS), and nonphosphorylated neurofilament protein (NPNFP). We found qualitative differences in the folding patterns of the IOpr among individuals and between the two sides of the brainstem. Quantification of IOpr volumes and indices of folding complexity, however, did not reveal consistent left-right differences in either parameter. Single-label immunohistochemistry showed that populations of neurons in the IOpr express CB, CR, NPNFP, and nNOS. Individual fibers labeled for PV, CB, CR, NPNFP, and nNOS were also found. There was individual variability in the numbers and density of stained neurons in the human IOpr; such variability was not seen in other brainstem nuclei. These data are consistent with, and complement, earlier studies showing a dramatic age-related increase in lipofuscin and decrease in RNA in the human IOpr. The impact of these changes in the IOpr on cerebellar function is, however, not known. PMID- 21630473 TI - Candidate molecular mechanisms for establishing cell identity in the developing retina. AB - In the developing nervous system, individual neurons must occupy appropriate positions within circuits. This requires that these neurons recognize and form connections with specific pre- and postsynaptic partners. Cellular recognition is also required for the spacing of cell bodies and the arborization of dendrites, factors that determine the inputs onto a given neuron. These issues are particularly evident in the retina, where different types of neurons are evenly spaced relative to other cells of the same type. This establishes a reiterated columnar circuitry resembling the insect retina. Establishing these mosaic patterns requires that cells of a given type (homotypic cells) be able to sense their neighbors. Therefore, both synaptic specificity and mosaic spacing require cellular identifiers. In synaptic specificity, recognition often occurs between different types of cells in a pre- and postsynaptic pairing. In mosaic spacing, recognition is often occurring between different cells of the same type, orhomotypic self-recognition. Dendritic arborization can require recognition of different neurites of the same cell, or isoneuronal self-recognition. The retina is an extremely amenable system for studying the molecular identifiers that drive these various forms of recognition. The different neuronal types in the retina are well defined, and the genetic tools for marking cell types are increasingly available. In this review we will summarize retinal anatomy and describe cell types in the retina and how they are defined. We will then describe the requirements of a recognition code and discuss newly emerging candidate molecular mechanisms for recognition that may meet these requirements. PMID- 21630475 TI - The expression pattern of polycomb group protein Ezh2 during mouse embryogenesis. AB - The Polycomb group (PcG) family proteins are required for the stability of homeotic selector genes and other genes related to the regulation of mammalian development through their roles in the modulation of chromatin domains. Among them, the mammalian enhancer zeste homologue 2 (Ezh2) contributes to the transcriptional repression of these genes. Previous studies tracked the Ezh2 expression at cDNA and mRNA levels during mouse development. However, little information is known about the expression patterns of Ezh2 at the protein levels. In this study, the embryos (E6.5-E18.5) obtained through timed matings of strain Kunming mice were inserted into paraffin blocks. Tissue microarrays were constructed and followed by subsequent immunohistochemical staining. The positive cells were identified and scored based on both the percentage of stained cells and their staining intensities. Ezh2 protein expression was found throughout the embryonic tissues including the nerves, intestine epithelial, liver, pancreas, renal tubule, and lungs. Its expression level was higher at early embryonic developmental stages. However, the nerve fibers and myocardium showed weak or no immunostaining reactivities. Ezh2 protein was moderately expressed in the nuclei of renal tubule epithelial cells at E14.5. In contrast, it was weakly expressed in the fetal kidneys at E18.5 and the protein was localized in the cytoplasm of the renal tubule epithelial cells. Our data confirmed that Ezh2 protein was expressed in mouse embryos and its expression exhibited tissue specificity and dependence on the stages of embryo development. thus providing new information helpful for understanding the possible roles of Ezh2 in embryogenesis. PMID- 21630479 TI - Systematic review of design and effects of splints and exercise programs in hand osteoarthritis. AB - OBJECTIVE: To describe and evaluate the design and effects of splints and exercise programs in hand osteoarthritis. METHODS: Controlled trials identified through systematic literature reviews were included. Design of splints and exercise programs were evaluated according to existing recommendations and classification systems. The risk of bias was assessed by 2 independent reviewers and effects were summarized descriptively or by meta-analyses. RESULTS: Twelve trials were included in the review: 7 assessed the effect of splints, 3 the effect of exercises, and 2 a combination of splints and exercises. The results revealed a great variety in the design of splint and exercise programs. A meta analysis of the 2 randomized trials with low risk of bias demonstrated that splints significantly reduced hand pain at short-term (<3 months) and long-term (>=3 months) followup, with a standardized mean difference of 0.37 (95% confidence interval [95% CI] 0.03, 0.71) and 0.80 (95% CI 0.45, 1.15), respectively. Further, results from single trials indicated that hand exercises may reduce pain and increase range of motion and strength, while a combination of splints and daily exercises may reduce pain and stiffness and improve function. CONCLUSION: There is consistent evidence that splints reduce hand pain, but limited evidence for the effects of hand exercises and a combination of hand exercises and splints in hand osteoarthritis. PMID- 21630480 TI - Fine needle aspiration cytology in sclerosing stromal tumor of the ovary: a series of three cases. AB - Sclerosing stromal tumor (SST) of the ovary is a distinct subtype of sex cord stromal tumor, which is derived from the ovarian stroma and, in turn, from the sex cords of the embryonic gonads. It is a benign tumor, predominantly, unilateral in nature and mostly involves the right side of the ovary. Histopathological features are characteristic and are well described in the literature; however to the best of our knowledge fine needle aspiration cytology (FNAC) has not been described previously. We present FNAC findings in a series of three cases of SST and discuss the cytomorphological features. We have described Call-Exner like bodies in one of our cases and hence, suggest that these may not be an exclusive feature of granulosa cell tumor. PMID- 21630481 TI - Crush intraoperatory analysis in craniopharyngioma. AB - Intraoperative smear cytology provides a rapid and reliable intraoperative diagnosis and guidance to the neurosurgeon during surgical resection and lesion targeting. It also helps the surgeon to monitor and modify the approach at surgery. The aim of this article was a clinicopathological and cytomorphological intraoperative crush smear correlation in craniopharyngioma. Thirty craniopharyngiomas were included in this study. Twenty-seven cases were adaCP and only three cases were papCP. This series included 16 (53%) males and 14 (47%) females adult patients, aged from 15 to 86 years (median, 49 year). Two cases were frank errors, 12 cases showed partial correlation, 5 cases showed incomplete typing of the cell type, and 7 cases discrepancy in type of tumors. The percent error was 14%. Correlations with clinical details and radiological findings were helpful in improving the accuracy rate. Smear technique is a fairly accurate, relatively safe, rapid, simple, easily reproducible, and cost-effective tool to diagnose brain tumors. Smear cytology is of great value in intraoperative consultation of central nervous system pathology. The cytological aspects and smear patterns disclose important complementary diagnostic information for the histopathological examination. PMID- 21630482 TI - Screening for cervical neoplasia: a community-based trial comparing Pap staining, human papilloma virus testing, and the new bi-functional Celldetect(r) stain. AB - Although cytological screening for cervical neoplasia has lowered mortality rates, current screening methods are plagued by sub-optimal sensitivity and/or specificity. The purpose of this study was to compare the performance of the new CellDetect(r) staining technology as a potential screening tool. This initial, non-blinded study, utilized samples are taken at a community-based clinic. The diagnostic results using CellDetect(r) were compared with the performance of Pap staining and human papilloma virus (HPV) testing on the same material, as well as the follow-up biopsies. These data were statistically analyzed in terms of sensitivity, specificity, predictive value (N.P.V and P.P.V), and inter-observer agreement. Bi-functional CellDetect(r) staining revealed morphological details and tinctorial properties that permitted recognition of neoplasia even at low magnification. Performance-wise, CellDetect(r) demonstrated non-inferiority for all statistical parameters to both Pap and HPV tests. Importantly, superior sensitivity compared with Pap staining was observed, as well as higher specificity than HPV testing with near equivalent sensitivity. We conclude that CellDetect(r) is a promising approach to early detection of cervical cancer because of its bi-functional capabilities that afford high sensitivity and specificity. The data suggest that this new methodology warrants further and more extensive clinical evaluation. PMID- 21630483 TI - Elastofibroma: cytomorphologic, histologic, and radiologic findings in five cases. AB - Elastofibroma is a rare benign, solid, ill-defined and slow-growing soft tissue lesion that typically arises between the tip of scapula and chest wall of elderly. Fine needle aspiration (FNA) is a commonly used technique to definitively diagnose the condition. However, the collagenous nature of the mass often results in a paucicellular FNA smear, resulting in a false-negative report. Herein, we report cytomorphologic, histological, and radiologic findings in five cases. Five cases of elastofibroma were retrospectively retrieved from the cytopathology archives of a teaching hospital. The cytological material was obtained by CT-guided (n = 2) and ultrasound-guided (n = 3) FNA. Smears were stained with Diff-Quik and Papanicolaou stains. Elastic stain and mucicarmine were reviewed in one case. Radiologic images and medical records and histology of all cases were reviewed. There were four patients (two men and two women). Their ages ranged from 64 to 84 (mean = 71.25 years). All tumors were subscapular (n = 2 right side, n = 1 left side, and one bilateral). Tumor sizes ranged from 3.3 to 7.5 cm in greatest dimension. Aspirated material was hypocellular in all cases, consisting of scattered uniform spindle cells, mature adipocytes, and collagen. Petaloid globules and serrated and braid-like linear elastic fibers were haphazardly scattered. The characteristic petaloid globules typical of elastofibroma in aspirated material can be inadvertently mistaken or overlooked sincethe samples are hypocellular. Awareness of cytomorphologic features of elastofibroma and the typical clinical setting will permit an accurate diagnosis and eliminate the need for preoperative biopsy. PMID- 21630484 TI - Rosai-Dorfman disease: A case report with extranodal thyroid involvement. AB - Rosai-Dorfman Disease (RDD) or Sinus Histiocytosis with Massive Lymphadenopathy (SHML) is a rare disorder typically manifesting as enlarged lymph nodes with or without systemic involvement. These cases are often clinically misdiagnosed as lymphoma. Recognising this entity to distinguish it from lymphoma and other causes of histiocytosis is important because of different treatment modalities for these disorders. Extranodal involvement is also common, often with a particular predilection for the head and neck region. We report a rare case of Rosai-Dorfman Disease with both nodal and extranodal involvement in a 33-year-old woman. The patient had bilateral cervical lymphadenopathy and diffuse thyroid enlargement. Thyroid gland involvement in RDD diagnosed on fine needle aspiration cytology (FNAC) has rarely been reported in literature. FNAC is a useful and reliable tool for the diagnosis of RDD and the biopsy can be avoided in these patients, thus reducing inconvenience to patients. PMID- 21630485 TI - Diagnosis and subclassification of lymphomas and non-neoplastic lesions involving mediastinal lymph nodes using endobronchial ultrasound-guided transbronchial needle aspiration. AB - INTRODUCTION: The value of endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) has been established for staging mediastinal lymph nodes in lung carcinoma patients with radiologically enlarged lymph nodes, but its utility for evaluation of primary lymph node disorders is not well defined. The objective of this study was to evaluate the usefulness of EBUS-TBNA with on-site assessment and triage of sample for multiple ancillary techniques, for the diagnosis and subclassification of lymphomas and non-neoplastic lesions involving mediastinal lymph nodes. METHODS: One hundred and twenty consecutive patients who underwent EBUS-TBNA between January 2008 and August 2009 were reviewed. The final cytological diagnosis was based on air-dried Romanowsky and alcohol-fixed Papanicolaou stained direct smears, immunohistochemistry, immunophenotyping, and fluorescence in situ hybridization (FISH). RESULTS: A total of 38 cases were included in this study consisting of eight reactive lymphoid hyperplasia, 20 granulomatous lymphadenitis (17 non-necrotizing and 3 necrotizing granulomatous inflammations), 3 Hodgkin lymphomas and 7 non-Hodgkin lymphomas (1 small lymphocytic lymphoma (SLL), 1 SLL with scattered Reed-Sternberg cells, 1 marginal zone lymphoma, and 4 large B cell lymphomas). Cultures performed in 13 cases were negative for AFB and fungi. Immunophenotyping and immunohistochemistry for MIB1 in six cases, and FISH in five cases provided necessary information for subclassification. CONCLUSIONS: EBUS-TBNA is a minimally invasive procedure which provides sufficient sample for definitive primary diagnosis and classification of malignant lymphoma and granulomatous inflammation in patients with mediastinal lymphadenopathy. Rapid on-site specimen assessment is invaluable for appropriate assignment of sample to ancillary studies. PMID- 21630486 TI - Ultra-stable phosphoglucose isomerase through immobilization of cellulose-binding module-tagged thermophilic enzyme on low-cost high-capacity cellulosic adsorbent. AB - One-step enzyme purification and immobilization were developed based on simple adsorption of a family 3 cellulose-binding module (CBM)-tagged protein on the external surface of high-capacity regenerated amorphous cellulose (RAC). An open reading frame (ORF) Cthe0217 encoding a putative phosphoglucose isomerase (PGI, EC 5.3.1.9) from a thermophilic bacterium Clostridium thermocellum was cloned and the recombinant proteins with or without CBM were over-expressed in Escherichia coli. The rate constant (kcat ) and Michaelis-Menten constant (Km ) of CBM-free PGI at 60 degrees C were 2,765 s(-1) and 2.89 mM, respectively. PGI was stable at a high protein concentration of 0.1 g/L but deactivated rapidly at low concentrations. Immobilized CBM (iCBM)-PGI on RAC was extremely stable at ~60 degrees C, nearly independent of its mass concentration in bulk solution, because its local concentration on the solid support was constant. iCBM-PGI at a low concentration of 0.001 g/L had a half-life time of 190 h, approximately 80-fold of that of free PGI. Total turn-over number of iCBM-PGI was as high as 1.1*10(9) mole of product per mole of enzyme at 60 degrees C. These results suggest that a combination of low-cost enzyme immobilization and thermoenzyme led to an ultra stable enzyme building block suitable for cell-free synthetic pathway biotransformation that can implement complicated biochemical reactions in vitro. PMID- 21630487 TI - Gas-permeable membranes and co-culture with fibroblasts enable high-density hepatocyte culture as multilayered liver tissues. AB - To engineer reliable in vitro liver tissue equivalents expressing differentiated hepatic functions at a high level and over a long period of time, it appears necessary to have liver cells organized into a three-dimensional (3D) multicellular structure closely resembling in vivo liver cytoarchitecture and promoting both homotypic and heterotypic cell-cell contacts. In addition, such high density 3D hepatocyte cultures should be adequately supplied with nutrients and particularly with oxygen since it is one of the most limiting nutrients in hepatocyte cultures. Here we propose a novel but simple hepatocyte culture system in a microplate-based format, enabling high density hepatocyte culture as a stable 3D-multilayer. Multilayered co-cultures of hepatocytes and 3T3 fibroblasts were engineered on collagen-conjugated thin polydimethylsiloxane (PDMS) membranes which were assembled on bottomless frames to enable oxygen diffusion through the membrane. To achieve high density multilayered co-cultures, primary rat hepatocytes were seeded in large excess what was rendered possible due to the removal of oxygen shortage generally encountered in microplate-based hepatocyte cultures. Hepatocyte/3T3 fibroblasts multilayered co-cultures were maintained for at least 1 week; the so-cultured cells were normoxic and sustained differentiated metabolic functions like albumin and urea synthesis at higher levels than hepatocytes monocultures. Such a microplate-based cell culture system appears suitable for engineering in vitro miniature liver tissues for implantation, bioartificial liver (BAL) development, or chemical/drug screening. PMID- 21630488 TI - Process development and cGMP manufacturing of a recombinant ricin vaccine: an effective and stable recombinant ricin A-chain vaccine-RVEcTM. AB - Ricin is a potent toxin and a potential bioterrorism weapon with no specific countermeasures or vaccines available. The holotoxin is composed of two polypeptide chains linked by a single disulfide bond: the A-chain (RTA), which is an N-glycosidase enzyme, and the B-chain (RTB), a lectin polypeptide that binds galactosyl moieties on the surface of the mammalian target cells. Previously (McHugh et al.), a recombinant truncated form of RTA (rRTA1-33/44-198 protein, herein denoted RVEaTM) expressed in Escherichia coli using a codon-optimized gene was shown to be non-toxic, stable, and protective against a ricin challenge in mice. Here, we describe the process development and scale-up at the 12 L fermentation scale, and the current Good Manufacturing Practice (cGMP)-compliant production of RVEcTM at the 40 L scale. The average yield of the final purified bulk RVEcTM is approximately 16 g/kg of wet cell weight or 1.2 g/L of fermentation broth. The RVEcTM was >99% pure by three HPLC methods and SDS-PAGE. The intact mass and peptide mapping analysis of RVEcTM confirmed the identity of the product and is consistent with the absence of posttranslational modifications. Potency assays demonstrated that RVEcTM was immunoprotective against lethal ricin challenge and elicited neutralizing anti-ricin antibodies in 95-100% of the vaccinated mice. PMID- 21630489 TI - [Application of thrombolytic therapy of ischemic stroke in the Russian Federation]. AB - To study the efficacy and safety of thrombolytic therapy (TLT) with the recombinant tissue plasminogen activator (rt-PA) in stroke, we treated 691 patients in 48 clinical units using systemic or selective TLT. Safety and high efficacy of TLT was shown: the three-months fatality rate was 18.2%, the symptomatic hemorrhage transformation rate related to clinical worsening was 6.1%. The good functional recovery (scores 0 or 1 on the modified Rankin scale) was observed in 48.6% of patients. PMID- 21630490 TI - [Rehabilitation of poststroke patients]. AB - The medical and social significance of acute cerebrovascular disturbances in the Russian Federation is considered. Purpose and tasks of early rehabilitation of patients with stroke are presented, its technology is described. Non-drug and pharmacological methods of rehabilitation are presented. The efficacy of neuromidin in the early rehabilitation of patients with a stroke is shown. PMID- 21630492 TI - Proceedings of the 15th Interdisciplinary Toxicology Conference TOXCON 2010. September 6-10, 2010. High Tatras, Stara Lesna . Slovakia. PMID- 21630491 TI - [The prognostic significance of fibrinogen concentration in patients with ischemic stroke without thrombolytic treatment]. AB - Recent studies showed that the increase of fibrinogen preceded not only thrombolytic events but also the development of intracerebral hemorrhage. The objective of the present study was to determine the threshold levels of fibrinogen concentration for the prognosis of ischemic stroke complications and, particularly, hemorrhagic transformation. Fibrinogen concentrations in patients with acute cerebral hemispheric stroke without thrombolytic treatment were examined. The lethal outcome and hemorrhagic transformation in patients during the hospital stay were selected as prognosis criteria. Four prognostic levels associated with the size of ischemic lesion, age and concomitant diseases were revealed. The high probability of the absence of complications regardless of the lesion size was observed when fibrinogen concentrations ranged between 200 and 350 mg/dL, the risk of hemorrhagic transformation with lethal outcome was higher in the range from 351 to 430 mg/dL and fibrinogen concentrations above 560 mg/dL predicted the high risk of lethal outcome. PMID- 21630493 TI - Pressure necrosis of ala nasi by Sengstaken-Blackemore tube. PMID- 21630494 TI - Impact of storage conditions on genetic analysis or viral load determination in clinical specimens. AB - BACKGROUND: Storage and shipment conditions of clinical specimens affect the quality of nucleic acids and may interfere with molecular analysis. The aim of our study was to verify whether blood storage at room temperature affects single nucleotide polymorphisms analysis; moreover, we analysed the consequences of serum storage at 4 degrees C on viral load determination of hepatitis B and C viruses. METHODS: For single nucleotide polymorphism screening, genomic DNA was extracted from EDTA whole blood samples stored at room temperature for different times, quantified photometrically, and Factor V Leiden point mutation analysis was performed. For viral load determination, serum samples with medium or low viremias were stored at +4 degrees C for different times and analysed by Cobas AmpliPrep/Cobas TaqMan tests for hepatitis B virus (HBV) DNA or hepatitis C virus (HCV) RNA. RESULTS: While mutation analysis was successfully performed on all samples tested, serum storage at +4 degrees C of HBV- and HCV-infected sera decreased viral load, in particular for low viremias. CONCLUSIONS: Storage of blood samples at room temperature up to 1 month does not affect the feasibility of genetic analysis, while serum storage at +4 degrees C affects viral load. PMID- 21630495 TI - Proceedings of the 3rd Croatian Congress on Side Effects of Psychotropics. March 23-27, 2011. Rovinj, Croatia. PMID- 21630496 TI - Political tussle continues over US health-care reform. PMID- 21630497 TI - Australian scientists lobby for antimicrobial oversight body. PMID- 21630499 TI - Retraction. Pharmacokinetics and clinical pharmacodynamics of the new propofol prodrug GPI 15715 in volunteers. PMID- 21630498 TI - The art of the demographic dividend. PMID- 21630500 TI - Retraction. Comparative pharmacokinetics and pharmacodynamics of the new propofol prodrug GPI 15715 and propofol emulsion. PMID- 21630501 TI - Retraction. AQUAVAN injection, a water-soluble prodrug of propofol, as a bolus injection: a phase I dose-escalation comparison with DIPRIVAN, Part 1: pharmacokinetics. PMID- 21630502 TI - Retration. AQUAVAN injection, a water-soluble prodrug of propofol, as a bolus injection: a phase I dose-escalation comparison with DIPRIVAN, Part 2: pharmacodynamics and safety. PMID- 21630503 TI - Brian Druker and Charles Sawyers receive the 2011 ASCI/Stanley J. Korsmeyer Award. PMID- 21630504 TI - Erratum: Purification of a toxic cysteine protease produced by pathogenic Aeromonas hydrophila isolated from rainbow trout. AB - An extracellular lethal toxin produced by Aeromonas hydrophila strain RT860715K originally isolated from diseased rainbow trout (Oncorhynchus mykiss) was purified by using Fast Protein Liquid Chromatography system with hydrophobic interaction chromatography and anion exchange columns. The toxin was a cysteine protease, inhibited by L-cysteine, iodoacetic acid, N-ethylamleimide, P chloromercuibenzene-sulfonic acid and N-alpha-p-tosyl-1-lysine-chloromethyl ketone (TLCK), and showed maximal activity at pH 6.0. The molecular weight of the purified enzyme proved to be 94 kDa as estimated by SDS-PAGE. In addition, the toxin was also completely inhibited by HgCl2 but partially inhibited by ethylenediamine tetraacetic acid (EDTA) and CuCl2. Both the extracellular products of Aeromonas hydrophila RT860715K and the purified protease were lethal to rainbow trout (weighing 18 g) with LD50 values of 2.87 and 0.93 MUg protein g 1 fish body weight, respectively. The addition of L-cysteine completely inhibited the lethal toxicity of the purified protease, indicating that this cysteine protease was a lethal toxin produced by the bacterium. PMID- 21630505 TI - Preventive health care, not examination. PMID- 21630506 TI - Reflections on then and now. PMID- 21630507 TI - Recent advances in the synthesis of morphine and related alkaloids. AB - Morphine, an alkaloid isolated from the opium poppy, has been widely used as an analgesic, and has been a fascinating synthetic target of organic chemists. After the first total synthesis reported in 1952, a number of synthetic studies toward morphine have been reported, and findings obtained in such studies have greatly contributed to the progress of synthetic organic chemistry as well as medicinal chemistry. This review provides an overview of recent studies toward the total synthesis of morphine and related alkaloids. Work reported in the literature since 2004 will be reviewed. PMID- 21630508 TI - Synthesis of 14-alkoxymorphinan derivatives and their pharmacological actions. AB - Among opioids, morphinans play an important role as therapeutically valuable drugs. They include pain relieving agents such as naturally occurring alkaloids (e.g. morphine, codeine), semisynthetic derivatives (e.g. oxycodone, oxymorphone, buprenorphine), and synthetic analogs (e.g. levorphanol). Currently used opioid analgesics also share a number of severe side effects, limiting their clinical usefulness. The antagonist morphinans, naloxone and naltrexone are used to treat opioid overdose, opioid dependence, and alcoholism. All these opioid drugs produce their biological actions through three receptor types, mu, delta, and kappa, belonging to the G-protein-coupled receptor family. Considerable effort has been put forward to understand the appropriate use of opioid analgesics, while medicinal chemistry and opioid pharmacology have been continuously engaged in the search for safer, more efficacious and nonaddicting opioid compounds, with the final goal to reduce complications and to improve patient compliance. Toward this goal, recent advances in chemistry, ligand-based structure activity relationships and pharmacology of 14-alkoxymorphinans are reviewed in this chapter. Current developments of different structural patterns of 14 alkoxymorphinans as research tools and their potential therapeutic opportunities are also summarized. PMID- 21630509 TI - Nonpeptidic delta (delta) opioid agonists and antagonists of the diarylmethylpiperazine class: what have we learned? AB - The discovery of the selective delta (delta) opioid agonists SNC 80 and BW373U86, which possess a diarylmethylpiperazine structure unique among opioids, represented a major advance in the field of delta-opioid ligands. Extensive research has recently been performed to uncover the structure-activity relationships (SAR) of this class of ligands, thereby providing valuable tools for the pharmacological characterization of the delta opioid receptor. This review focuses on the SAR of this unique series of ligands, and provides an overview of the various chemical routes that have been developed and optimized through the years to allow the syntheses of these ligands on a multigram scale. The search for selective delta opioid agonists and antagonists, as well as for those with mixed opioid agonist properties with potential therapeutic value, continues. Several questions regarding the interaction at the molecular level of diphenylmethylpiperazine derivatives and related analogs with opioid receptors and in particular with the delta opioid system still remain unanswered. Indeed, the development and pharmacological characterization of novel nonpeptidic delta opioid ligands remains an active area of research, as it may provide a better understanding of the role of this receptor in multiple disease states and disorders. PMID- 21630510 TI - Synthesis of novel basic skeletons derived from naltrexone. AB - We will describe eight interesting reactions using naltrexone derivatives. Almost all these reactions are characteristic of naltrexone derivatives, and can lead to the synthesis of many novel skeletons that provide new interesting pharmacological data. Some of the new reactions that were found with naltrexone derivatives were expanded into general reactions. For example, the reaction of 6alpha-hydroxyaldehyde derived from naltrexone led to the oxazoline dimer and the 1,3,5-trioxazatriquinane skeleton (triplet drug); this reaction was applied to general ketones which were converted to alpha-hydroxyaldehydes, followed by conversion to dimers and trimers, as described in Sect. 7. PMID- 21630511 TI - 3D-pharmacophore identification for kappa-opioid agonists using ligand-based drug design techniques. AB - A selective kappa-opioid receptor (KOR) agonist might act as a powerful analgesic without the side effects of micro-opioid receptor-selective drugs such as morphine. The eight classes of known KOR agonists have different chemical structures, making it difficult to construct a pharmacophore model that takes them all into account. Here, we summarize previous efforts to identify the pharmacophore for kappa-opioid agonists and propose a new three-dimensional pharmacophore model that encompasses the kappa-activities of all classes. This utilizes conformational sampling of agonists by high-temperature molecular dynamics and pharmacophore extraction through a series of molecular superpositions. PMID- 21630512 TI - Muscular dystrophies: a way forward. Proceedings of the Ninth Neurobiology of Diseases in Children Symposium. October 13, 2009. Louisville, Kentucky, USA. PMID- 21630514 TI - Chemistry of opioids. Preface. PMID- 21630513 TI - Electromigration techniques in pharmaceutical and biomedical analysis. Festschrift in honour of Professor Jacques Crommen on the occasion of his 60th birthday. PMID- 21630515 TI - Opioids in preclinical and clinical trials. AB - Since 1952, when Gates determined the stereo structure of morphine, numerous groups have focused on discovering a nonnarcotic opioid drug. Although several natural, semisynthetic, and synthetic opioid ligands (alkaloids and peptides) have been developed in clinical studies, very few were nonnarcotic opioid drugs. One of the most important studies in the opioid field appeared in 1976, when Martin and colleagues established types of opioid receptors (these are now classified into mu, delta, and kappa types). Later, Portoghese discovered a highly selective mu type opioid receptor antagonist, beta-funaltrexamine. This led to the finding that the mu type opioid receptor was correlated to drug dependence. Consequently, delta, and particularly kappa, opioid agonists were expected to lead to ideal opioid drugs. Moreover, opioid antagonists were evaluated for the treatment of symptoms related to undesirable opioid system activation. In this chapter, we provide a short survey of opioid ligands in development and describe the discovery of the two most promising drugs, TRK-851 and TRK-820 (nalfurafine hydrochloride). PMID- 21630516 TI - 14-Amino-4,5-epoxymorphinan derivatives and their pharmacological actions. AB - 14-Hydroxy-7,8-dihydromorphinone (oxymorphone) and its derivatives (oxycodone, naloxone, naltrexone) have become among the most important clinical agents to have been produced from opium. 14-Aminocodeinone and its 7,8-dihydro and morphinone derivatives are of more recent origin thanks to the work of Professor Gordon Kirby and his collaborators. The 14-amino parent compounds have proved of limited interest but their 14-acylamino- and 14-alkylamino derivatives have been extensively studied. The 4'-substituted cinnamoylamino-17-cyclopropylmethyl-7,8 dihydronormorphinones, C-CAM and M-CAM are the best available selective MOR irreversible antagonists and the related dihydrocodeinone MC-CAM, 4'-chloro cinnamoylamino-17-cyclopropylmethyl-7,8-dihydronorcodeinone, is a long-acting MOR partial agonist with extended MOR-pseudoirreversible antagonist activity that could be a candidate for pharmacotherapy of opiate abuse/dependence. PMID- 21630517 TI - Synthesis of neoclerodane diterpenes and their pharmacological effects. AB - Salvinorin A is a neoclerodane diterpene that has been shown to be an agonist at kappa opioid receptors. Its unique structure makes it an attractive target for synthetic organic chemists due to its seven chiral centers and diterpene scaffold. This molecule is also interesting to pharmacologists because it is a non-serotonergic hallucinogen, and the first opioid ligand discovered that lacks a basic nitrogen. There have been several total synthesis approaches to salvinorin A, and these will be detailed within this chapter. Additionally, research efforts have concentrated on structure modification of the salvinorin A scaffold through semi-synthetic methods. Most modifications have focused on the manipulation of the acetate at C-2 and the furan ring. However, chemistry has also been developed to generate analogs at the C-1 ketone, the C-4 methyl ester, and the C-17 lactone. The synthetic methodologies developed for the salvinorin A scaffold will be described, as well as specific analogs with interesting biological activities. PMID- 21630518 TI - Twin and triplet drugs in opioid research. AB - Twin and triplet drugs are defined as compounds that contain respectively two and three pharmacophore components exerting pharmacological effects in a molecule. The twin drug bearing the same pharmacophores is a "symmetrical twin drug", whereas that possessing different pharmacophores is a "nonsymmetrical twin drug." In general, the symmetrical twin drug is expected to produce more potent and/or selective pharmacological effects, whereas the nonsymmetrical twin drug is anticipated to show both pharmacological activities stemming from the individual pharmacophores (dual action). On the other hand, nonsymmetrical triplet drugs, which have two of the same pharmacophores and one different moiety, are expected to elicit both increased pharmacological action and dual action. The two identical portions could bind the same receptor sites simultaneously while the third portion could bind a different receptor site or enzyme. This review will mainly focus on the twin and triplet drugs with an evaluation of their in vivo pharmacological effects, and will also include a description of their pharmacology and synthesis. PMID- 21630519 TI - [The role of human papillomavirus typization and cytology in early detection of relapse of cervical intraepithelial neoplasia]. AB - BACKGROUND/AIM: Female patients who underwent ceratin treatment forms of cervical intraepithelial neoplasia (CIN) are at five times greater risk for disease relapse in comparison to the rest of female population. The aim of the study was to investigate validity of human papillomavirus (HPV) typization and cytology in detection of relapse. METHODS; The prospective clinical investigation included 35 patients with relapse and 30 ones without it after adequate treatment of cervical intraepithelial neoplasia. HPV typization using PCR methods and cytological test (conventional Pap smear) were performed in all the patients. Validation of tests applied was performed by determining their sensitivity, specificity, and positive and negative predictive value. RESULTS: More severe degrees of CIN relapse occur significantly more often in patients which remain HPV positive despite of the treatment. The patients which remain positive on HPV type 18 or, on both HPV types 18 and 16, have more often CIN relapses of more severe degree in relation to those just positive on HPV type 16. HPV typization has higher predictive value for diagnosis of the rezidual disease in older patients. Sensitivity of HPV typization for diagnosis all CIN relapse degrees is 68.57%, for more severe degrees (HSIL and MIC) 90.47%, specificity is 93.33%, while positive and negative predictive values are 90.47% and 93.53%, respectively. Sensitivity of cytology for diagnosis of more severe CIN relapses is 80.95%. HPV typization used along with cytology ofters the highest sensitivity (95.23%). CONCLUSION: Both tests, HPV typization and Pap smear, offer satisfactory sensitivity and high specificity in detection of relapse, parcticularly those with more severe degree. The highest sensitivity in detection of CIN relapse is obtained by using both tests. PMID- 21630520 TI - [Hormonal status in patients with advanced prostatic cancer on the therapy with androgen blockade]. AB - BACKGROUND/AIM: Hormone suppression therapy is used in men with advanced prostate cancer improving chances of longer survival. The aim of this study was to investigate the influence of androgen blockades on testosterone and luteinizing hormone (LH) values in patients with locally advanced and metastatic prostatic cancer. METHODS: The study included a total of 60 patients out of which 45 with prostatic cancer divided into 3 subgroups based on the type of the applied treatment protocol: 15 patients on monotherapy with luteinizing-releasing hormone (LH-RH) agonists (group I), 15 patients on total androgen blockade (group II) and 15 patients on monotherapy with antiandrogen (group III)). The control group consisted of 15 patients with benign prostatic hyperplasia. In all the patients, values of testosteron, LH and prostate-specific antigen (PSA) were monitored initially, as well as 3 and 6 months after the treatment protocol introduction. RESULTS: In the patients of the groups I, II and III, values of testosterone decreased after three months by 95.58%, 95.72%, and 67%, respectively. The difference was significant (p < 0.01). Between the values after three and six months there was no significant difference in these groups of participants. Testosterone values were significantly higher in the patients of the group III in both analyses. Comparing the values between the groups III and I, as well as those of the groups III and II, a significant difference was found after three and six months of the therapy (p < 0.01). There was a difference in testosterone values between the groups I and II after 3 and 6 months, but not significant. All types of the applied treatment protocols in the therapy of prostatic cancer significantly decreased the values of LH compared to the basal ones. CONCLUSION: Total androgen blockade and LH-RH agonists are more effective in lowering testosterone values (to castration values) compared to the antiandrogen monotherapy, where testosterone values stay above the castration level. This therapy approach has advantages, since it decreases testosterone values providing better therapy response. There is a difference in testosterone values, but not significant, when total androgen blockade and monotherapy with LH-RH agonists are administered. Registered lower basal values of LH in all patients with prostatic cancer open the possibility to introduce LH as a new additional, significant marker in diagnosis of this neoplasm. PMID- 21630521 TI - [Heritability of bipolar affective disorder--family study]. AB - BACKGROUND/AIM: Bipolar affective disorder is mental disorder with polygenic type of heredity. Heritability--relation between genetic and environmental variance is used to estimate the level of influence of genetic variance to phenotype variance. Study results show decreasing trend in the value of heritability of bipolar affective disorder, thus indicating that this disorder is a complex behavioral threshold characteristic. Therefore, the aim of this study was to estimate the contribution of genetic variance to phenotype variance of bipolar affective disorder, i.e. to estimate heritability of this disorder. METHODS: By the use of a questionnaire, 80 patients with over crossed threshold for bipolar affective disorder were asked for functional information about the members of their families belonging to the first degree of relation (fathers, mothers and full-sibs). By using "Applet for calculating heritability for threshold traits (disease)", and regression analysis, heritability of bipolar affective disorder as well as its statistical significance, were estimated (chi2 test). RESULTS: Heritability and relationship of genetic and environmental variance of bipolar affective disorder is 0.2 with statistically significant difference from zero (p < 0.001). CONCLUSION: The estimated contribution of genetic variance to phenotype variance of bipolar affective disorder is low being 20%, while the contribution of environmental variance is 80%. This result contributes to the understanding of bipolar affective disorder as a complex behavioral threshold trait. PMID- 21630522 TI - [Computer-aided technologies in diagnostics and therapy of impacted teeth]. PMID- 21630523 TI - [Delayed angioedema during therapy with angiotensin-converting enzyme inhibitors]. AB - INTRODUCTION: Angiotensin-converting enzyme inhibitors are leading cause of drug induced angioedema, with incidence of 0.1 to 0.2%. The angioedema is not of immune nature; in predisposed individuals it is caused by accumulation of vasoactive mediators due to reduced activity of angiotensin-converting enzyme. CASE REPORT: We presented a 63-year old male patient suffering from hypertension and chronic obstructive pulmonary disease, who had developed two episodes of angioedema during a 5-year long therapy with enalapril. The first episode happened after three, and the second after five years of the therapy. On both occasions, the patient was admitted to the hospital and tracheotomy was avoided in the last moment. CONCLUSION: Long-term therapy with angiotensin-converting enzyme inhibitors could be associated with delayed angioedema, especially in patients with inflammation of airways caused by infection or chronic irritation. PMID- 21630524 TI - [Overview of psychiatry in Japan--from the viewpoint of psychiatric emergencies]. PMID- 21630525 TI - [Conditions for effective psychiatric emergency care in the framework of regional health care]. PMID- 21630526 TI - [Positive and negative aspects of therapy, treatment and assistance in rehabilitation of patients with drug dependence--from the acute care to assistance to normal return to the local community]. PMID- 21630527 TI - [Goal for emergency psychiatric care]. PMID- 21630528 TI - [Diagnosis and psychoanalytical care of personality disorders]. PMID- 21630529 TI - [Treatment of PTSD by applying prolonged exposure therapy--evidence found in Japan and its application at the care of victims]. PMID- 21630530 TI - [Concept of dysfunctions in daily activities of dementia victims and a need for scientific approach]. PMID- 21630531 TI - [BAL30072--an iron complexing beta lactam antibiotic against multidrug-resistant gram negative bacteria]. PMID- 21630532 TI - [The flexibility of the protein neuraminidase opens new possibilities for drug development]. PMID- 21630533 TI - [Vascular damage in cancer therapy]. PMID- 21630534 TI - [Flu outbreaks. Influenza viruses on the way from animal to man]. PMID- 21630535 TI - [Seasonal pathogen epidemics and global pandemics. Influenza virus: biology and pathogenicity]. PMID- 21630536 TI - [Pathogenesis, symptoms and treatment of virus influenza]. PMID- 21630537 TI - [The new (threatening?) influenza pandemics]. PMID- 21630538 TI - [Flu vaccines]. PMID- 21630539 TI - [Flu vaccine production in hens's eggs]. PMID- 21630540 TI - [The production of influenza vaccines from MDCK cell cultures]. PMID- 21630541 TI - [M2 inhibitors and neuraminidase inhibitors]. PMID- 21630542 TI - [How safe was oseltamivir care in the flu pandemic?]. PMID- 21630543 TI - Surgery in Vukovar war. PMID- 21630544 TI - [RNA interference test for discovery of new targets]. PMID- 21630545 TI - [Strategies for development of new influenza medication]. PMID- 21630546 TI - State-based coverage solutions: the California Health Benefit Exchange. AB - California was the first state to create its own health insurance exchange after the passage of the Affordable Care Act. Because of its front-runner status and the sheer size of its coverage expansion, California's choices will have implications for other states as they address difficult issues, including minimizing adverse selection, promoting cost-conscious consumer choice, and seamlessly coordinating with public programs. California took advantage of the flexibility in the federal health reform law to create an exchange that will function as an active purchaser in the marketplace; take significant steps to combat adverse selection both against and within the exchange, including requiring all insurers to sell all tiers of products and making exchange participation a condition of selling catastrophic plans; and allow community based health plans to develop commercial offerings for the exchange. This brief examines these decisions, which will provide a roadmap for other states as they set up their exchanges. PMID- 21630547 TI - [Prevention of visceral worms and schistosomiasis]. PMID- 21630548 TI - [The modified Lichtenstein technique for complex inguinal hernia repair--how I do it]. AB - The Lichtenstein technique is modified for solving complex groin hernias such as huge hernias with massive transversal fascia destruction associated with the increased intraabdominal pressure or recurrent hernias with the destroyed Poupart's ligament. Whilst these hernias are usually managed by preperitoneal techniques (open or laparoscopic) under general or regional anesthesia, as an "inpatient" procedure, they can be solved applying a modified Lichtenstein technique, most frequently under local anesthesia, as an "out-patient" procedure. The modifications of Lichtenstein technique include the foIlowing: a) lateral movement and fixation of the lower corner of the mesh, caudally to the tubercle, by 20-30 degrees in relation to its lower border, fully protecting the medial triangle (direct inguinal recurrence prevention); b) fixation of the lower border of the mesh by a running "U" suture to both Poupart's and Coopers's ligaments, from the tubercle to the femoral vein, fully protecting the femoral triangle (femoral recurrence prevention); c) the lower mesh border fixation by a running suture, 2-3 cm laterally to the internal inguinal ring, together with the "locking" of the internal inguinal ring by two interrupted sutures, one fixing the superior mesh tail to the inferior one--cranial to the spermatic cord, 1-1,5 cm medially to the Poupart's ligament, and the other fixing the lower border of the superior mesh tail and the lower border of the inferior mesh tail to the inferior part of the Poupart's ligament, 1 cm cranially and laterally to the preceding suture, fully protecting the lateral triangle (indirect inguinal recurrence prevention). One thousand eighteen patients with 1236 (unilateral 800, bilateral 218) inguinal hernias were electively operated on by the modified Lichtenstein technique between January 2003-January 2011. All operations were performed by a single surgeon. One hundred and thirty (10.5%) hernias were recurrent following one or more tension or tension-free repairs, and 203 (16.4%) hernias had a > or = 5 cm hernial defect. In seven hundred and twentyfour (71.1%) patients, the operation was performed under local, in 271 (26.6%) under general, and in 23 (2.3%) under regional anesthesia, while 635 (62.4%) patients were operated on an "out-patient" basis, and 383 (37.6%) on an "in-patient" basis. The ASA score was: 388 ASA I, 450 ASA II, 153 ASA III, and 27 ASA IV. The mean stay at a day surgery unit was 2.5 (2-8) hours, and the mean hospital stay was 1.6 (1 10) days. During the mean follow-up of 37 (1-96) months, the rate of complications was: 23 (1.86%) haematoma, 5 (0.4%) seroma, 5 (0.4%) wound infections, 6 (0.48%) ischaemic orcihitis, 2 (0.16%) testicle atrophy, 1 (0.08%) disejaculation, 3 (0.24%) hydrocoella, 21 (1.7%) pain, and 2 (0.16%) recurrence. There were 6 reoperations due to the complications. The modified Lichtenstein technique performed usually under local anesthesia as "a day-case" procedure is a good solution for challenging groin hernias. PMID- 21630549 TI - Dynamic MR imaging of kidneys perfused with EOB-Gd-DTPA. AB - Gandolinium-ethoxybenzyl-diethylenetriamine pentaacetic acid (Gd-EOB-DTPA) is a hepatobiliary contrast agent for MRI. It was reported that Gd-EOB-DTPA is useful to detect liver tumors and differentially diagnose benign and malignant pathologies in the liver. Since Gd-EOB-DTPA partially accumulates in the hepatocytes and bile via various transporters after intravenous injection, signal intensity in the liver increases on T1-weighted images. The signal intensity of the liver after Gd-EOB-DTPA injection depends on the Gd-EOB-DTPA uptake by hepatocytes and bile excretion. It is known tha the Gd-EOB-DTPA accumulating in the kidney is excreted to the urine through glomerular filtration. Because Gd DTPA is concentrated in the renal tubules after being filtered at the Bowman's capsule, and since it is neither secreted nor reabsorbed the concentrating and diluting function of the renal tubules can be studied by imaging techniques. since renal function can be evaluated with Gd-EOB-DTPA can also be used to evaluate renal function. Eith the development of MRI equipment and rapid imaging techniques, temporal resolution had improved greatly. However, no previous study has been carried out on renal function using Gd-EOB-DTPA dynamic MR study that was correlated with estimated glomerular filtration rate (eGFR) and stage of chronic kidney disease (CKD) in the Japan Association of chronic kidney disease initiative. PMID- 21630550 TI - Possible role of the scintigraphic estimation of the relative liver perfusion in the diagnosis and therapy of liver carcinomas. AB - BACKGROUND: The aim of the study is evaluation of the possible role of the scintigraphic estimation of the relative liver perfusion in diagnosis and the choice of treatment of liver carcinomas. MATERIAL AND METHODS: Hepatic perfusion index was obtained by dynamic scintigraphy in 126 patients. RESULTS: In the control group values did not differ from the value in the patients with benign tumors (p > 0.05). However, in hepatocellular carcinoma and liver metastases of different tumors, HPI values were significantly decreased in comparison to controls and benign tumors (p < 0.01), but they didn't differ between themselves (p > 0.05). The values were especially low in the patients with malignant diseases in the liver accosciated with vascular disturbances in the portal system. CONCLUSION: HRA could be easily done during the different conventional nuclear medicine methods. It can be an useful method for the assessment of different degrees of hemodynamic alterations in portal system, for differential diagnosis of benign and malignant liver tumors, as well as for assessment of the liver tissue and tumor perfusion, which might be helpful in the decision making for the undertaking of intraarterial (radionuclide, chemotherapy etc.) therapy. PMID- 21630551 TI - The analysis of some factors that influence on serum PSA level in localized prostate cancer patients: mathematical model. AB - INTRODUCTION: Serum prostate specific antigen (PSA) concentration in localized prostate cancer (PCa) patients depends of numerous factors related to tumor, prostate and the patient. Tumor factors include tumor volume, localization, growth rate and aggressiveness. Prostate factors include prostate volume, the activity of dihydrotestosterone (DHT) and PSA synthesis. Host factors include androgenic status and total blood volume. OBJECTIVE: to discuss all the possible factors that influence on serum PSA levels in localized PCa patients. To present the mathematical model that predicts serum PSA concentration in localized PCa patients. METHODS: Mathematical model is based on the hypothesis that tumor related PSA molecules are released from the epithelial prostatic cells around the tumor, so called "Destruction Zone" around the tumor (DZ). The amount of PSA is dependent of DZ volume. Moreover, DZ volume depends on tumor volume, tumor localization in the prostate and tumor growth rate. RESULTS: The study offers the wide spectrum of PSA values, dependent on tumor, prostate and patient characteristics. These values are comparable with the empirical results in the literature, especially if tumor-volume doubling time of three years is entered into the calculations. CONCLUSION: Although with some limitations, this mathematical model can explain the great variety of PSA serum concentrations in PCa patients. PMID- 21630552 TI - Ureteral fistulae after kidney transplantation: experience with 224 cases. AB - Ureteral fistulae after kidney transplantation represent major early urological complications with reported incidence from 1.2% to 12% in large series. The aim of the study is to establish the incidence, types and ureteral fistula related morbidity and lethality rates, by donor type. From 1995. to 2001, a total of 224 kidney transplantations (171 from living and 53 from cadaveric donor) have been performed at the Institute of Urology and Nephrology in Belgrade. Mean patients age was 36,67 years (11-64; SD = 10.69). Ureteral fistulae appeared only after living donor transplantation in a total of five patients (2.2%) (p > 0.05). In all patients open fistula repair was performed. Two patients had recidive ureteral fistula after primary and after secondary open repair. Following the third open repair one patient had lethal outcome. Ureteral fistulae after kidney transplantation still remain challenging urological problem with considerable morbidity and lethality rates. PMID- 21630553 TI - Helicobacter pylori infection in various groups of patients studied, estimated by 14C-urea breath test. AB - The aim of this study was the detection of helicobacter pylori (HP) infection and estimation of the clinical validity and the accuracy of the 14C-urea breath test in the groups of patients studied. A total of 248 patients with gastric diseases were examined. There were 38 patients with gastric ulcer, 41 with duodenal ulcer, 43 with gastroduodenitis erosiva, 26 with hiatus hernia, 36 with gastric carcinoma and 64 patients with gastritis. There were 103 true positive (TP), 139 true negative (TN), 4 false negative (FN) and 2 false positive (FP) patients. There was no significant difference in the incidence of the HP infection between the groups of patients studied (p > 0.05). Sensitivity of the method was 96.3%, specificity 98.6%, positive predictive value 98.1%, negative predictive value 97.2% and accuracy 97.6%. Our results point out that the method is very accurate for the detection of HP infection. PMID- 21630554 TI - Usage of a Trans-Obturator-Tape (T.O.T.) "outside-in" approach in surgical treatment of female stress urinary incontinence. AB - OBJECTIVE: The aim of the study was to analyzed the efficacy and safety of a new minimally invasive surgical procedure using the Trans-Obturator-Tape with"outside in" approach for treatment female stress urinary incontinence. PATIENTS AND METHODS: 31 women with stress urinary incontinence (SUI) associated with urethral hypermobility, underwent the T.O.T. procedure (March 2010 to January 2011). 5 patients were previously operated for incontinence. Mean age was 59 years (37 80). 10 patients were having mixed incontinence. A non-elastic, polypropylene tape was placed under the mid-urethra. The surgical placement technique utilises a trans-obturator percutaneous approach. All patients underwent post-operative clinical examination, cough-stress test (full bladder), uroflowmetry, and post voiding residual assessment. RESULTS: Mean follow-up was 5 months (1-9). At 6 months follow-up 96.7% of the patients were completely cured. The overall peri operative complication rate was 6.4% with no vascular, nerve or bowel injury. One patients (3.4%) had post-operative urinary retention. CONCLUSION: The present study confirms the results obtained by the instigator of the technique, E. Delorme, and allows us to consider T.O.T. as an effective and safe technique for the treatment of female stress urinary incontinence. PMID- 21630555 TI - The importance of physical therapy in the postoperative period after total cystectomy of bladder carcinoma. AB - The aim of this study is to evaluate the results and complications after radical cystectomy due to carcinoma of the bladder and to point out the significance of post-operative physical treatment and rehabilitation of these patients. MATERIAL AND METHOD: In the period of 3 years (2007-2010), at the Urological Clinic in Belgrade, we performed 195 total cystectomies for invasive bladder carcinoma with the use of different types of urinary diversion. The operation was performed in 162 men (83%) and 33 women (17%). RESULTS: Survival, complications and postoperative recovery was dependent on the type of urinary diversion which was used, stage of disease and general condition of patients before surgery. The worst result was achieved in patients who underwent ureterocutaneostomy and the complications were represented in 30% of patients. In the group of patients where the ileal conduit was applied, complications were recorded in 10% of patients, while mortality was 5%. In the group of patients where the continent urinary diversion was performed, complications were recorded in 5% of patients in mind of stecoral fistulas, urinary fistulas and ileus. CONCLUSION: The timely application of the physical therapy and rehabilitation in these patients is of great importance, because it reduces complications and allows faster recovery and release from the hospital. PMID- 21630556 TI - Get documentation improvement in order or risk major trouble. AB - Clinical documentation improvement is taking on a bigger importance as payers tighten reimbursement rules, public reporting of hospital data increases, and a variety of auditors are scrutinizing hospital records, making it imperative that the medical record accurately reflects the patient's condition and the services he or she received. Assemble a multidisciplinary team to create and monitor your documentation improvement program. Develop a training program for your documentation improvement staff. Encourage collaboration between the documentation staff and case management. Look for opportunities for improvement and track your outcomes. PMID- 21630557 TI - Language is key in clinical documentation. AB - Medical language and coding language are not the same, which means that physicians often write what they learned in medical school to describe a patient's condition: if it doesn't contain the proper language, the coders can't accurately code the chart. Educate physicians to use appropriate language to reflect the condition of the patient. Look for hints in documentation by other clinicians that the physician documentation might not be accurate. Ensure that documentation is complete to indicate major complication/comorbidity rates (CCs or MCCs). PMID- 21630558 TI - Documentation staff can alleviate CM duties. AB - Hospitals need a separate, dedicated clinical documentation staff to fill in the gap between what is documented in the chart and what is coded. Case managers have too many other duties to take on this complex task. Candidates should be able to work independently, have good communication skills, good rapport with doctors. They need to have clinical credibility and understand the MS-DRG system, public reporting, and ICD-9 codes. Staff needs extensive training, preferably by an outside consultant. PMID- 21630559 TI - Case management report cards--keep it simple, keep it clear. PMID- 21630560 TI - Pre-billing review will improve mortality index. AB - At Stony Brook University and Medical Center in Stony Brook, NY, clinical documentation specialists conduct pre-billing reviews of the charts of patients who die in the hospital to ensure that risk of mortality is accurately represented. Staff reviews charts of all patients with an expected mortality rate of 3 or lower. Review is pre-billing because publicly reported mortality data is based on billing information. When physician query is returned, coders can recode the chart. PMID- 21630561 TI - Reorganization adds staff and improvements. AB - Wake Forest Baptist Medical Center in Winston-Salem, NC, reorganized its clinical documentation program and added more staff after working with a consultant to determine opportunities for improvement. Staff members are assigned by service and work as a team with case managers. The hospital added four clinical documentation consultants. Clinical documentation consultants, coders, physicians received training over four weeks. PMID- 21630562 TI - Notifying patients of their right to complain. AB - A proposed rule issued by the Centers for Medicare and Medicaid Services would require hospitals and other providers to formally notify Medicare beneficiaries about their right to complain to the state Medicare Quality Improvement Organization (QIO). Rule applies to outpatients as well as inpatients. Process is likely to tie into value-based purchasing. Case managers should take proactive steps to deal with complaints. PMID- 21630563 TI - ED connects patients to better venues of care. PMID- 21630564 TI - Novel mutation in TGA stop-codon of bovine SIX6 gene. AB - As a transcriptional regulatory gene of the SIX family, SIX6 (also known as OPTX2, SIX9), probably affects pituitary development and secretion of hormones, suggesting that this gene is a potential candidate gene for studying association with growth trait in animals. Therefore, this study is first of all focused on detecting sequence variations in a bovine SIX6 gene and on its effects on growth traits in 1087 cattle from five Chinese cattle breeds using DNA sequencing and HhaI-ACRS-PCR methods. Herein, a novel mutation (NC_007308: g 2015T > C) in the TGA stop-codon of a bovine SIX6 gene was found, which leads to an ORF shift and extension of the encoded protein for four amino acids (Arg223-Gln224-Arg225 Val226). The frequency of allele "C" varied from 0.255 (Chinese Holsteins) to 0.614 (Hasake). They all were in the Hardy-Weinberg equilibrium except Jiaxian Red and Hasake cattle. Using amino acid sequence alignment and online prediction software, a new helix in the C-terminal domain of the mutated bovine SIX6 protein was revealed, which possibly affects pituitary development and hormone secretion. So, relationship analysis between this polymorphism and growth traits in the Nanyang breed was carried out based on a proper linear model. Although no statistically significant associations were observed (P > 0.05), the presented work preliminarily demonstrated a novel mutation in the TGA stop-codon which extends the spectrum of genetic variations of the bovine SIX6 gene and might be of interest in terms of its association with other biophysical and biochemical indexes. PMID- 21630565 TI - Genomic analysis of silkworm microRNA promoters and clusters. AB - MicroRNAs (miRNAs) are endogenous single-stranded RNAs of 18-22 nt in length, which can regulate the complementary mRNAs at the post-transcriptional level by cleavage or repression of translation of the target mRNAs. Studies have shown that the majority of animal miRNAs are transcribed from independent transcription units, and someare transcribed together with their host genes. However, the nature of the primary transcript of intergenic miRNAs remains unknown. Silkworm (Bombyx mori) miRNAs are representative of those of the Lepidoptera insects and many of them are conserved in Caenorhabditis elegans and other animal species. To date, little is known about the transcriptional regulation of silkworm miRNA genes. We performed the genomic analysis on the silkworm miRNA transcripts around the promoter region including the transcription start site (TSS) and the TATA box, and on the organization of the miRNA cluster. In 73 pre-miRNAs from the silkworm 131 promoters were detected via a bioinformatics approach. Among them the portion of non-conserved promoters is greater than that of the conserved ones. The genomic organization of pre-miRNAs of the silkworm was globally analyzed and it was determined that 11 of them we reorganized into five clusters. Sequence alignment showed that paralogs existed for some of the miRNAs in the cluster. These results may increase the understanding of the specific sequences upstream of the pre-miRNAs and of the functional implications of miRNA clusters in the silkworm. PMID- 21630566 TI - Identification of exotic genetic components and DNA methylation pattern analysis of three cotton introgression lines from Gossypium bickii. AB - The impact of alien DNA fragments on plant genome has been studied in many species. However, little is known about the introgression lines of Gossypium. To study the consequences of introgression in Gossypium, we investigated 2000 genomic and 800 epigenetic sites in three typical cotton introgression lines, as well as their cultivar (Gossypium hirsutum) and wild parents (Gossypium bickii), by amplified fragment length polymorphism (AFLP) and methylation-sensitive amplified polymorphism (MSAP). The results demonstrate that an average of 0.5% of exotic DNA segments from wild cotton is transmitted into the genome of each introgression line, with the addition of other forms of genetic variation. In total, an average of 0.7% of genetic variation sites is identified in introgression lines. Simultaneously, the overall cytosine methylation level in each introgression line is very close to that of the upland cotton parent (an average of 22.6%). Further dividing patterns reveal that both hypomethylation and hypermethylation occurred in introgression lines in comparison with the upland cotton parent. Sequencing of nine methylation polymorphism fragments showed that most (7 of 9) of the methylation alternations occurred in the noncoding sequences. The molecular evidence of introgression from wild cotton into introgression lines in our study is identified by AFLP. Moreover, the causes of petal variation in introgression lines are discussed. PMID- 21630567 TI - Anti-inflammatory effect of the alpha-melanocyte stimulating hormone in animal eyes undergoing extracapsular lens extraction. AB - In this study anti-inflammatory effects of the alpha-melanocyte stimulating hormone (alpha-MSH) on ocular inflammation caused by extracapsular lens extraction (ECLE) have been investigated and the potential mechanism of an anti inflammatory effect is discussed. Pigmented rabbit eyes after ECLE were treated locally with alpha-MSH, dexamethasone, diclofenac, or saline 4 times a day (q.i.d.) for 4 weeks. The inhibitory effect of alpha-MSH on infiltrating cells in the aqueous humor (AqH) was almost twice as good as that of dexamethasone or diclofenac for 3 days, 1 week, and 2 weeks after the operation. The eyes of Sprague-Dawley rats were treated with an intravenous injection of alpha-MSH or saline immediately after ECLE. Six hours postoperatively, the iris/ciliary body exhibited increased expression of TNF-alpha and IL-6 mRNAs, which were significantly decreased after alpha-MSH treatment. The number of activated NF kappa B (NFkappaB)-positive cells in the iris/ciliary body was also significantly reduced by the alpha-MSH treatment. These results suggested that alpha-MSH could effectively reduce ocular inflammation after ECLE, and the potential mechanism for this is by down-regulating the expression of proinflammatory cytokines and inhibiting the NFkappaB-dependent signaling pathway. PMID- 21630568 TI - New antibacterial therapeutics and strategies. AB - Studies on new antibacterial therapeutics and strategies are currently being conducted in many microbiological, pharmaceutical and biochemical laboratories. The antibacterial activity of plant-derived compounds as well as silver and gold nanoparticles is the subject of this minireview. The application of photodynamic therapy is also discussed. PMID- 21630569 TI - Cloning, expression and identification by immunohistochemistry of humanized single-chain variable fragment antibody against hepatitis C virus core protein. AB - Expression of single-chain variable fragment (scFv) antibodies on the surface of bacteriophage is widely used to prepare antibodies with pre-defined specificities. A phage antibody library containing the gene for scFv antibody against Hepatitis C virus core protein was panned with core protein immobilized on microtiter plate wells. After five rounds of panning 60 phage clones specific to core protein were obtained and one selected clone was sequenced. It was found that the specifically detected antigen consists of 774bp and is capable of encoding 257 amino acids in the patients but not in healthy persons. PMID- 21630570 TI - The occurrence and comparative phenotypic characteristics of Staphylococcus spp. from healthy and diseased, household and shelter dogs, based on routine biochemical diagnostic methods. AB - To determine the staphylococcal colonization pattern in healthy and diseased dogs, living in two particular environments, a number of microbiological samples were taken. Overall, twenty dogs, either healthy or with infected skin lesions, were examined. In each case bacterial swabs were collected from the nasal mucosa, ear, perineum, lumbo-sacralis triangle, and from the infection sites if such were present. A total number of 104 isolates representing different staphylococcal species were isolated and identified using routine biochemical methods applied in diagnostic laboratories. Among 17 isolated staphylococcal species, Staphylococcus intermedius was the most common species isolated from both healthy or diseased dogs living either in animal shelter or household environments. The pattern of Staphylococcus sp. colonization differs considerably for animals living in the two tested habitats. In particular, S. aureus MRSA and MSSA isolates were detected only in infected skin lesion samples from animals that dwelled in the animal shelter. As could be expected, S. intermedius was found to be a predominant causative agent in canine skin infections. In our study, we demonstrated that S. intermedius in its carrier-state, inhabits mainly the mucosal membrane of the nasal vestibule. It was also found in the samples taken from the skin, the lumbo-sacralis triangle and perineum, but was rarely isolated from the ears. PMID- 21630571 TI - Comparison of different PCR methods for detection of Brucella spp. in human blood samples. AB - For detection of Brucella species by PCR four DNA extraction methods and four targets were compared using pure culture of Brucella melitensis and the best conditions were applied in clinical samples. It was found that the MagNA Pure LC method was the most efficient and sensitive method showing a positive PCR reaction with DNA extracted from as low as 25 and 100 CFU suspended in one ml blood and one ml water, respectively. Detection of Brucella spp. by conventional PCR was investigated using four different targets. The results indicated that The B4-B5 amplification method was the most sensitive one as it could amplify DNA extracted from as a low as 25 and 100 CFU/ml suspended in one ml water and blood, respectively. Furthermore real-time PCR was able to detect Brucella using DNA extracted from as low as 50 CFU/ml blood and 15 CFU/ml water, respectively. The best and optimum detection conditions were applied to the clinical samples. Evaluation of conventional PCR assays on blood specimens confirmed 72% of the results obtained by conventional blood culture methods with a specificity of 95%, while serum samples had a sensitivity of 54% and specificity of 100%. Real-time PCR was generally found to be more sensitive and specific for detecting Brucella spp. in blood and serum samples compared to conventional PCR. The real-time PCR done on blood specimens confirmed 77.5% of the results obtained by conventional blood culture methods with specificity of 100%, while 60% of serum samples were found to be positive with specificity of 100%. These results suggest that serum and blood analysis by conventional and real time PCR is a convenient and safe method for rapid and accurate diagnosis of brucellosis. PMID- 21630572 TI - Antibiofilm activity of selected plant essential oils and their major components. AB - The aim of the study was to examine the antibiofilm activity of selected essential oils (EO): Lavandula angustifblia (LEO), Melaleuca alternifolia (TTO), Melissa officinalis (MEO) and some of their major constituents: linalool, linalyl acetate, alpha-terpineol, terpinen-4-ol. Biofilms were formed by Staphylococcus aureus ATCC 29213 and Escherichia coli NCTC 8196 on the surface of medical biomaterials (urinary catheter, infusion tube and surgical mesh). TTC reduction assay was used for the evaluation of mature biofilm eradication from these surfaces. Moreover, time-dependent eradication ofbiofilms preformed in polystyrene 96-well culture microplates was examined and expressed as minimal biofilm eradication concentration (evaluated by MTT reduction assay). TTO, alpha terpineol and terpinen-4-ol as well as MEO, showed stronger anti-biofilm activity than LEO and linalool or linalyl acetate. Among the biomaterials tested, surgical mesh was the surface most prone to persistent colonization since biofilms formed on it, both by S. aureus and E. coli, were difficult to destroy. The killing rate studies of S. aureus biofilm treated with TTO, LEO, MEO and some of their constituents revealed that partial (50%) destruction of 24-h-old biofilms (MBEC50) was achieved by the concentration 4-8 x MIC after 1 h, whereas 2-4 x MIC was enough to obtain 90% reduction in biomass metabolic activity (MBEC90) after just 4 h of treatment. A similar dose-dependent effect was observed for E. coli biofilm which, however, was more susceptible to the action of phytochemicals than the biofilms of S. aureus. It is noteworthy that an evident decrease in biofilm cells metabolic activity does not always lead to their total destruction and eradication. PMID- 21630573 TI - Competitiveness of Rhizobium leguminosarum bv. trifolii strains in mixed inoculation of clover (Trifolium pratense). AB - Rhizobium leguminosarum by. trifolii (Rlt) establishes beneficial root nodule symbiosis with clover. Twenty Rlt strains differentially marked with antibiotic resistance markers were investigated in terms of their competitiveness and plant growth promotion in mixed inoculation of clover in laboratory experiments. The results showed that the studied strains essentially differed in competition ability. These differences seem not to be dependent on bacterial multiplication in the vicinity of roots, but rather on complex physiological traits that affect competitiveness. The most remarkable result of this study is that almost half of the total number of the sampled nodules was colonized by more than one strain. The data suggest that multi-strain model of nodule colonization is common in Rhizobium-legume symbiosis and reflects the diversity ofrhizobial population living in the rhizosphere. PMID- 21630574 TI - Selection and adaptation of Saccharomyces cerevisae to increased ethanol tolerance and production. AB - A total of 24 yeast strains were tested for their capacity to produce ethanol, and of these, 8 were characterized by the best ethanol yields (73.11-8 1.78%). The most active mutant Saccharomyce s cerevisiae ER-A, resistant to ethanol stress, was characterized by high resistance to acidic (pH 1.0 and 2.0), oxidative (1 and 2% of H2O2), and high temperature (45 and 52 degrees C) stresses. During cultivation under all stress conditions, the mutants showed a considerably increased viability ranging widely from about 1.04 to 3.94-fold in comparison with the parent strain S. cerevisiae ER. At an initial sucrose concentration of 150 g/l in basal medium A containing yeast extract and mineral salts, at 300C and within 72 h, the most active strain, S. cerevisiae ER-A, reached an ethanol concentration of 80 g/1, ethanol productivity of 1.1 g/Il/h, and an ethanol yield (% of theoretical) of 99.13. Those values were significantly higher in comparison with parent strain (ethanol concentration 71 g/1 and productivity of 0,99 g/l/h). The present study seems to confirm the high effectiveness of selection of ethanol-resistant yeast strains by adaptation to high ethanol concentrations, for increased ethanol production. PMID- 21630575 TI - Cytotoxicity of Aspergillus fungi isolated from hospital environment. AB - The majority of mycotoxins produced by Aspergillus fungi are immunosuppressive agents, and their cytotoxicity may impair defense mechanisms in humans. The objective of the study was evaluation of the cytotoxicity of fungi isolated from an environment where inpatients with impaired immunity were present. The materials comprised 57 fungal strains: Aspergillus fumigatus, Aspergillus niger: Aspergillus ochraceus, Aspergillus flavus, Aspergillus versicolor and Aspergillus ustus isolated from hospital rooms in Cracow. The cytotoxicity of all the strains was evaluated using the MTT test (3-(4,5-dimethylthiazol-2-yl) 2,5 diphenyltetrazolium bromide). To emphasize the differences in cytotoxicity among the particular strains, variance analysis (ANOVA) and Tukey's difference test were used. Out of 57 Aspergillus strains tested, 48 (84%) turned out to be cytotoxic. The cytyotoxicity was high (+++) in 21 strains, mainly in A. fumigatus. The least cytotoxic were A. niger fungi, this being statistically significant (p<0,05). To protect a patient from the adverse effects of mycotoxins, not only his or her immunity status should be evaluated but also the presence of fungi in hospital environment and their cytotoxicity should be monitored (possible exposure). PMID- 21630576 TI - Production and partial characterization of high molecular weight extracellular alpha-amylase from Thermoactinomyces vulgaris isolated from Egyptian soil. AB - Optimizing production of alpha-amylase production by Thermoactinomyces vulgaris isolated from Egyptian soil was studied. The optimum incubation period, temperature and initial pH of medium for organism growth and enzyme yield were around 24 h, 55 degrees C and 7.0, respectively. Maximum alpha-amylase activity was observed in a medium containing starch as carbon source. The other tested carbohydrates (cellulose, glucose, galactose, xylose, arabinose, lactose and maltose) inhibited the enzyme production. Adding tryptone as a nitrogen source exhibited a maximum activity of alpha-amylase. Bactopeptone and yeast extract gave also high activity comparing to the other nitrogen sources (NH4CI, NH4NO3, NaNO3, KNO3, CH3CO2NH4). Electrophoresis profile of the produced two alpha amylase isozymes indicated that the same pattern at about 135-145 kDa under different conditions. The optimum pH and temperature of the enzyme activity were 8.0 and 60 degrees C, respectively and enzyme was stable at 50 degrees C over 6 hours. The enzyme was significantly inhibited by the addition of metal ions (Na+, Co2+ and Ca2+) whereas CI- seemed to act as activator. The enzyme was not affected by 0.1 mM EDTA while higher concentration (10 mM EDTA) totally inactivated the enzyme. PMID- 21630577 TI - Halomonas sp. nov., an EPA-producing mesophilic marine isolate from the Indian Ocean. AB - Marine samples from the Indian Ocean were used to isolate and characterize the organisms with respect to their fatty acid profiles. Six mesophilic isolates (MBRI 6, MBRI 8, MBRI 9, MBRI 10, MBRI 12 and MBRI 13) were obtained from three different water samples. They were i) Gram-negative, ii) catalase positive, iii) produced acid from glucose and maltose, iv) tolerated 5 to 15% NaCI v) except MBRI 9, showed pH tolerance in the range of 5.0 to 9.0 with optimum pH 7.0 to 8.0 v) grew well at 30 degrees C and were able to grow in the range of 15 to 45 degrees C. EPA, an essential omega-3 fatty acid, was produced by these isolates in the range of 12 to 60% at 30 degrees C. MBRI 12 was found to be a potential source as it produced 60% EPA. This isolate was further identified by partial 16S rDNA sequencing and phylogenetic analysis revealed that the strain belonged to Gammaproteobacteria and was closely related to Halomonas bolviensis (96% sequence similarity, 570 bp). Thus a new genus of Halomonas may be included in earlier reported EPA- producing prokaryotic genera affiliated to the Gammaproteobacteria. PMID- 21630578 TI - Yersinia species in the dunnock (Prunella modularis) in sub-alpine habitats of the Western Carpathians. AB - The study presents the prevalence of Yersinia species in dunnok Prunella modularis from the sub-alpine zone of the Western Carpathians. Bacteria were detected from cloacal and pharyngeal swabs from 97 specimens using PCR assay. Yersinia enterocolitica showed the highest prevalence (47.4%) from among the determined Yersinia species. Yersinia species (except Y frederiksenii) were detected more frequently in pharyngeal than cloacal samples. The highest prevalence of yersiniosis was detected in April (Yersinia spp. - 80%, Y. enterocolitica - 70%). No statistically differences were observed in the prevalence of Yersinia spp. between males and females and between juveniles and adult birds. Bacterial contamination did not affect body weight or tarsus length. PMID- 21630579 TI - First report of Serratia plymuthica causing onion bulb rot in Poland. AB - Specific bacterial disease symptoms were observed on onion bulbs in almost all regions in Poland. For the purpose of identification of agents causing disease, bacteria were isolated from the symptomatic plants. Their pathogenicity was confirmed by using pathogenicity test on onion scales. These bacteria were identified biochemically and molecularly as Serratia plymuthica. PMID- 21630580 TI - Assessment of predictors of mortality and severity in patients with colorectal perforation. AB - Colorectal perforation is a life-threatening disease with high mortality and morbidity. The correct and prompt diagnosis and accurate judgment of severity are necessary. We retrospectively investigated 30 patients with colorectal perforation to assess predictors of mortality and severity, and evaluated the usefulness of computed tomography (CT) for the initial diagnosis. The severity of peritonitis was assessed using clinical factors and Physiological and Operative Severity Score for the enUmeration of Mortality and morbidity (POSSUM) scores. Abdominal free air was detected by CT in 92% of patients with colorectal perforation, whereas only 36.6% showed evidence of abdominal free air by conventional radiography. The perforation site was correctly diagnosed in 14 of 25 cases (56%). Overall mortality was 16.7%. Survivors were younger than nonsurvivors, and POSSUM physiological and mortality scores were significantly lower for survivors compared with nonsurvivors. The amount of intraperitoneal soiling by large bowel content determined disease severity in terms of the need for postoperative respiratory management. In conclusion, CT is necessary for precise diagnosis, and the POSSUM score is helpful for the evaluation of disease mortality and severity. PMID- 21630581 TI - Isoflavonoid derivatives from Lophira alata stem heartwood. AB - Six isoflavonoid derivatives among which three are new have been isolated from the stem heartwood of Lophira alata. The structures were elucidated from spectroscopic and chemical evidences. Two have unusual carbon skeletons, possibly resulting from a variant of isoflavonoid biogenesis. The two compounds form the first members of a new subclass of flavonoid compounds which we call "isobiflavonoids". The presence of these isoflavonoid compounds in this plant of the Ochnaceae family has important chemotaxonomic implications since it modifies the botanic distribution of isoflavonoid compounds in non-leguminous plants. PMID- 21630582 TI - Synthesis of benzothiophene carboxamide derivatives and their pharmacological evaluation as potent antihypertriglyceridemic agents in rats. AB - Benzothiophene carboxamide derivatives of aminobenzophenone, aminopyridine, aminobenzimidazole, and aniline derivatives (compounds 1-9) were synthesized and compounds 3, 6, 7, 8, and 9 tested in vivo for their hypolipidemic activity. Compounds 1-8 were prepared adopting the fusion process at 130-150 degrees C between benzothiophene-2-carbonyl chloride and aminobenzophenones, aminopyridine, and anilines, respectively, and were obtained in high yield, while compound 9 was obtained from the reaction of benzothiophene acyl chloride with aminobenzimidazole in DMF at 160 degrees C. At a dose of 15 mg/kg body weight compounds 6, 7, and 9 significantly reduced plasma triglyceride levels in Triton WR-1339-induced hyperlipidemic rats in comparison to control rats. Furthermore, they significantly increased high-density lipoprotein levels. It is therefore reasonable to assume that compounds 6, 7, and 9 may have a promising potential in the treatment of hyperlipidemia and atherosclerosis. PMID- 21630583 TI - Synthetic 3-arylideneflavanones as inhibitors of the initial stages of biofilm formation by Staphylococcus aureus and Enterococcus faecalis. AB - The antimicrobial activity of twenty two synthetic flavonoids is reported. Among them three 3-arylideneflavanones, 2b, 2c, and 2i, were shown to be highly active against Staphylococcus aureus, S. epidermidis, and Enterococcus faecalis reference strains, with MIC (minimal inhibitory concentration) values ranging from 4.68 microg/ml (14.3 microM) to 37.5 microg/ml (119.7 microM). The synergy of oxacillin and vancomycin with 2c, evaluated as fractional inhibitory concentration index (FICI) was shown (against planktonic culture of S. aureus A3 and E. faecium 138/09 clinical strains). The presence of 2c in the culture medium diminished the initial adhesion of bacteria to an abiotic surface. Such an effect resulted in a decrease in biofilm formation during prolonged culture. Unfortunately, 2e failed to eradicate the S. aureus mature biofilm which was already preformed, however, decreased the number of live biofilm cells. The biofilm of E. faecalis was more susceptible to the action of 3-arylideneflavanone 2c than the S. aureus biofilm. The finding that 3-arylideneflavanones are lipophilic, cause bacterial aggregation, and influence the integrity of membranes making them permeable to SYTO 9/propidium iodide dyes may implicate the cytoplasmic membrane as a target site for these compounds activity. PMID- 21630584 TI - Chemical composition and biological activity of Laennecia schiedeana. AB - The chemical study of Laennecia schiedeana afforded three sterols, five diterpenes, five flavonoids, three caffeoyl derivatives of quinic acid, and two triterpenes. Evaluation of the cytotoxic activity of the extracts and isolated metabolites showed that 15-methoxy-16-oxo-15,16H-strictic acid was the most active compound [(15.05 +/- 2.2) microg/mL against U-251 cells]. In antibacterial assays the acetonic extract of leaves was the only active extract exhibiting its highest effect against the multiresistant Staphylococcus epidermidis (MIC 0.25 mg/mL). The anti-inflammatory activity observed was mild in the extracts and not relevant in the isolated compounds. PMID- 21630585 TI - Antibacterial activity of an oligosaccharide of native Paecilomyces sp. and its aminoglycosylated derivative. AB - This study reports the antibacterial activity of an oligosaccharide, prepared by partial acid hydrolysis of a native Paecilomyces sp. exopolysaccharide, and of its aminoglycosylated derivative, prepared by reductive alkylation of the oligosaccharide, against E. coli and S. aureus. PMID- 21630586 TI - Repellent and insecticidal activities of Melia azedarach L. against cotton leafworm, Spodoptera littoralis (Boisd.). AB - A crude acetone extract and oil of ripe fruits from Melia azedarach L. were evaluated against the 2nd and 4th instar larvae of Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae). Both oil and extract exhibited highly significant growth inhibition at all concentrations tested, while the oil of M. azedarach recorded higher insecticidal activity against both instars than the crude extract. GC-MS analysis of the oil revealed the presence of linoleic acid methyl ester, oleic acid methyl ester, and free oleic acid as the main components in addition to hexadecanol, palmitic acid, methyl esters of stearic acid and myristic acid. Fatty acids and their esters were not only the main constituents of essential oil from the ripe fruits of M. azedarach, but also mainly responsible for the insecticidal and growth inhibition activity against S. littoralis. PMID- 21630587 TI - Quantitative structure-activity relationship of Morita-Baylis-Hillman adducts with leishmanicidal activity. AB - A quantitative structure-activity relationship model for Morita-Baylis-Hillman adducts with leishmanicidal activities was developed which correlates molecular orbital energy and dipole with percentage in the promastigote stage. PMID- 21630588 TI - Effect of the method of preparation on the composition and cytotoxic activity of the essential oil of Pituranthos tortuosus. AB - The aerial parts of Pituranthos tortuosus (Desf.) Benth and Hook (Apiaceae), growing wild in Egypt, yielded 0.8%, 0.6%, and 1.5% (v/w) of essential oil when prepared by hydrodistillation (HD), simultaneous hydrodistillation-solvent (n pentane) extraction (Lickens-Nickerson, DE), and conventional volatile solvent extraction (preparation of the "absolute", SE), respectively. GC-MS analysis showed that the major components in the HD sample were beta-myrcene (18.81%), sabinene (18.49%), trans-iso-elemicin (12.90%), and terpinen-4-ol (8.09%); those predominent in the DE sample were terpinen-4-ol (29.65%), sabinene (7.38%), gamma terpinene (7.27%), and beta-myrcene (5.53%); while the prominent ones in the SE sample were terpinen-4-ol (15.40%), dill apiol (7.90%), and allo-ocimene (4E,6Z) (6.00%). The oil prepared in each case was tested for its cytotoxic activity on three human cancer cell lines, i.e., liver cancer cell line (HEPG2), colon cancer cell line (HCT116), and breast cancer cell line (MCF7). The DE sample showed the most potent activity against the three human cancer cell lines (with IC50 values of 1.67, 1.34, and 3.38 microg/ml against the liver, colon, and breast cancer cell lines, respectively). Terpinen-4-ol, sabinene, gamma-terpinene, and beta myrcene were isolated from the DE sample and subjected to a similar evaluation of cytotoxic potency; significant activity was observed. PMID- 21630589 TI - Lipid profiling of Synechococcus sp. PCC7002 and two related strains by HPLC coupled to ESI-(ion trap)-MS/MS. AB - The lipid profiles of Synechococcus sp. PCC7002 and two related 16S rDNA (99% identity) strains were established by a new method of high-performance liquid chromatography coupled to electrospray-mass spectrometry (HPLC-MS). Lipids were analysed in the positive and negative ionization mode, and fragmentation patterns are reported. No differences in the lipid profile between the three strains could be observed, but the relative content of some species differed. Major lipid species were found to be 1-octadecatrienoyl-2-hexadecanoyl-3-(6'-sulfo-alpha-D quinovosyl)-sn-glycerol [SQDG (18:3/16:0)] and 1-octadecatrienoyl-2-hexadecenoyl 3-beta-D-monogalactosyl-sn-glycerol [MGDG (18:3/16:1)]. Ten species of SQDG, six species of PG (phosphatidyl-glycerol), seven species of MGDG, and two species of DGDG (digalactosyl-diacyl-glycerol) were detected. A PG species (m/z 761) containing hydroxylinolenic acid or oxophytodienoic acid acyl ester (C18H32O3), and SQDG species containing C17:1 and C17:3 fatty acyl esters are reported for the first time in cyanobacteria. The method also allowed the separation of two pairs of closely related isobaric MGDG species (m/z 770 and m/z 772 in positive ionization). PMID- 21630590 TI - Darkness affects differentially the expression of plastid-encoded genes and delays the senescence-induced down-regulation of chloroplast transcription in cotyledons of Cucurbita pepo L. (Zucchini). AB - In contrast to differentiated leaves, the regulatory mechanisms of chloroplast gene expression in darkened cotyledons have not been elucidated. Although some results have been reported indicating accelerated senescence in Arabidopsis upon reillumination, the capacity of cotyledons to recover after dark stress remains unclear. We analysed the effect of two-days dark stress, applied locally or at the whole-plant level, on plastid gene expression in zucchini cotyledons. Our results showed that in the dark the overall chloroplast transcription rate was much more inhibited than the nuclear run-on transcription. While the activities of the plastid-encoded RNA polymerase (PEP) and nuclear RNA polymerase II were strongly reduced, the activities of the nuclear-encoded plastid RNA polymerase (NEP) and nuclear RNA polymerase I were less affected. During recovery upon reillumination, chloroplast transcription in the cotyledons was strongly stimulated (3-fold) compared with the naturally senescing controls, suggesting delayed senescence. Northern blot and dot blot analyses of the expression of key chloroplast-encoded photosynthetic genes showed that in contrast to psbA, which remained almost unaffected, both the transcription rate and mRNA content of psaB and rbcL were substantially decreased. PMID- 21630591 TI - Identification of species-diagnostic inter simple sequence repeat markers for ten Phyllanthus species. AB - Phyllanthus has been widely used in traditional medicine as an antipyretic, a diuretic, and to treat liver diseases and viral infections. Correct genotype identification of medicinal plant material remains important for the botanical drug industry. Limitations of chemical and morphological approaches for authentication have generated the need for newer methods in quality control of botanicals. In the present study, attempts were made to identify species diagnostic markers for ten Phyllanthus species using the inter simple sequence repeat-polymerase chain reaction (ISSR-PCR) fingerprinting method. PCR amplification using seven ISSR primers resulted in significant polymorphism among the populations from different species. P. angustifolius and P. urinaria showed monomorphic frequency of maximum (63.88%) and minimum (20.64%), respectively. Seventeen species-diagnostic markers were identified for seven species (P. acidus, P. emblica, P. fraternus, P. urinaria, P. rotundifolius, P. amarus, and P. angustifolius) while no marker was detected for P. reticulatus, P. nivosus, and P. virgulatus. A maximum of six species-diagnostic markers were identified for P. acidus and a minimum of only one of 755 bp was available for P. amarus. Among the seventeen markers, nine were present in all individuals of particular species. The species-specific differences in fragment numbers and sizes could be used as diagnostic markers to distinguish the Phyllanthus species quickly. PMID- 21630592 TI - Statistical optimization of the medium composition by response surface methodology to enhance schizophyllan production by Schizophyllum commune. AB - The response surface methodology (RSM) involving central composite design (CCD) was employed to optimize the fermentation medium for the cell growth and schizophllan production by Schizophyllum commune CGMCC 5.113 in submerged culture at pH 6.5 and 26 degrees C. The four variables involved in this study were glucose, yeast extract, ammonium nitrate, and magnesium sulfate. The statistical analysis of the results showed that, in the range studied, glucose and yeast extract had a highly significant effect on schizophyllan production. The optimal medium for schizophyllan production calculated from the regression model of RSM was as follows: glucose, 18 g/l; yeast extract, 0.5 g/l; NH4NO3, 0.48 g/l; and MgSO4, 0.05 g/l, with a predicted maximum schizophyllan production of 11.74 g/l. These predicted values were experimentally validated. The excellent correlation between predicted and measured values justifies the validity of the response model. The results of bioreactor fermentation also show that the optimized medium enhanced schizophyllan production (12.80 g/l) by S. commune in a 5-1 fermenter. PMID- 21630593 TI - Copper-binding peptides from human prion protein and newly designed peroxidative biocatalysts. AB - A previous work suggested that peptides from the histidine-containing copper binding motifs in human prion protein (PrP) function as peroxidase-like biocatalysts catalyzing the generation of superoxide anion radicals in the presence of neurotransmitters (aromatic monoamines) and phenolics such as tyrosine and tyrosyl residues on proteins. In this study, using various phenolic substrates, the phenol-dependent superoxide-generating activities of PrP-derived peptide sequences were compared. Among the peptides tested, the GGGTH pentapeptide was shown to be the most active catalyst for phenol-dependent reactions. Based on these results, we designed a series of oligoglycyl-histidines as novel peroxidative biocatalysts, and their catalytic performances including kinetics, heat tolerance, and freezing tolerance were analysed. PMID- 21630594 TI - Single-embryo metabolomics and systematic prediction of developmental stage in zebrafish. AB - Metabolites, the end products of gene expression in living organisms, are tightly correlated with an organism's development and growth. Thus, metabolic profiling is a potentially important tool for understanding the events that have occurred in cells, tissues, and individual organisms. Here, we present a method for predicting the developmental stage of zebrafish embryos using novel metabolomic non-target fingerprints of "single-embryos". With this method, we observed the rate of development at different temperatures. Our results suggest that this method allows us to analyse the condition, or distinguish the genotype, of single embryos before expression of their ultimate phenotype. PMID- 21630595 TI - New risks in executive pay. PMID- 21630596 TI - True partners. AB - Physician alignment is essential for quality improvement. Hospitals can turn independent docs into allies by giving them a voice in governance, management and strategic planning. PMID- 21630597 TI - Simple steps to better physician relations. PMID- 21630598 TI - Physician compensation requires enhanced board oversight. PMID- 21630599 TI - Drug shortage turns severe. AB - Limited production capacity and concerns about raw materials mean common medications are in short supply. How can your facility avoid a crisis? PMID- 21630600 TI - Trends in board performance. PMID- 21630601 TI - Standing by your CEO. PMID- 21630602 TI - Status quo: not an option. PMID- 21630603 TI - Medication safety. PMID- 21630604 TI - Patient adherence to oral anticancer drugs: an emerging issue in modern oncology. AB - The steady increase in the use of oral anticancer drugs in modern oncology has created a paradigm shift, challenging traditional attitudes towards cancer care and requiring new concepts of organization of oncology services. Important issues are the prolonged treatment period, management of toxicity, treatment adherence, reimbursement conditions and patient and family education. Although most patients generally prefer oral therapy over intravenous treatment for reasons of convenience, the daily use of oral anticancer drugs can be a challenging commitment for many patients. Reports on adherence and persistence among patients with cancer show that adherence ranges from 16% to 100%, depending on the type of therapy and the measurement/definition of adherence. Apart from demographic, disease and therapy related factors, the determinants that mostly influence (non )adherence are the satisfaction with care activities performed at the initiation of the drug treatment, and the perceived necessity of treatment. Therefore, patient education addressing these issues is considered the cornerstone of successful oral anticancer treatment. Studies examining the role of different health care providers in the pharmacotherapeutic care of patients with cancer, treated with oral anti-cancer drugs, support the need for a multidisciplinary approach to achieve a maximum benefit for the individual patient and consequently for the whole health system. Limiting adverse events and developing appropriate supportive care are only some aspects that need to be considered in this. PMID- 21630605 TI - Dermatological side effects of current and upcoming targeted therapies in oncology. AB - Targeted therapies are gaining field in oncology practice. Some of them are already well established, others are upcoming. They target cancer cells more selectively, therefore causing less collateral damage. Dermatologic side effects are common and sometimes class specific. The skin toxicity profile of EGFR inhibitors, MEK en Raf inhibitors, mTOR inhibitors, VEGF targeting molecules, multikinase inhibitors, the HER2 monoclonal antibody trastuzumab and the CTLA-4 monoclonal antibodies are discussed. When possible, a pathogenic mechanism and treatment options are described. PMID- 21630606 TI - Renal biopsy findings in Belgium: a retrospective single center analysis. AB - Renal biopsy is the definitive diagnostic test in patients with renal parenchymal disease. Renal biopsy registry is an important tool which can provide valuable data concerning early and correct epidemiological description and clinical correlations of renal diseases. Records of 326 adult renal biopsies performed at our hospital from January 1991 till the end of December 2006 were retrospectively examined. Overall, secondary glomerular diseases (SGD) were predominant (39.9%), followed by primary glomerular diseases (PGD) (30.4%), vascular diseases (13.2%) and TIN (6.7%). Total sclerosis of the kidney did not allow histopathological diagnosis in 5.8% of all biopsied kidneys. Focal and Segmental Glomerular Sclerosis (FSGS), IgA Nephropathy (IgAGN) and Minimal Change Disease (MCD) and Membranous Glomerulopathy (MGN) were the most common PGD, altogether representing 75.7% of all PGD. FSGS was the most frequent (30.3%), followed by IgAGN (21.2%), MCD (19.1%) and MGN in 15.1%. Vasculitis, HIVAN, diabetic nephropathy and amyloidosis were the most common SGD, altogether representing 90% of all SGD. Immune Mediated Glomerulonephritis (IMGN) were the most frequent (32.3%), followed by HIVAN (16.9%), diabetic nephropathy (14.6%) and amyloidosis (10%). Nephroangiosclerosis (benign and malignant nephroangiosclerosis) was the most frequent vascular nephropathy responsible for 79% of all vascular diseases. Thrombotic microangiopathy was seen in 9.3% and atherothrombotic disease in 7% of all vascular diseases. Concerning tubular diseases, chronic TIN accounted for 63.6% of all tubular diseases, followed by light chain-cast nephropathy (22.7%) and acute TIN (13.6%). Because of lack of material, 3.4% of all biopsies could not be analyzed. These data demonstrate that the distribution of biopsy-proved renal diseases in a Belgian population of the Brussels area is strongly influenced by the indications of renal biopsy. Harmonization of these indications might reflect with more accuracy the actual incidence of different nephropathies in a given population. Nation and worldwide renal biopsy registers are important to follow patterns of renal diseases in different populations. This information is important not only for health organizations in order to plan health budget but also for helping clinicians to provide a better care to patients. PMID- 21630607 TI - Diagnostic yield of syncope investigation (initiated) in the emergency department: a pilot study. AB - OBJECTIVE: To determine the diagnostic yield of investigations in patients presenting to the emergency department (ED) with syncope. To determine the cause of the syncope. PATIENTS AND METHODS: We retrospectively identified consecutive patients presenting to the ED who underwent investigations for syncope at a 1900 bed university teaching hospital during 4 months. From the medical records we abstracted clinical information, results of testing and the cause of syncope. RESULTS: A total of 117 patients participated in the study. The mean age was 57 year (range 6-93 year), and 45% were male. Diagnostic procedures most often performed were blood analysis, ECG, chest X-ray and Holter monitoring (respectively in 94.1%, 88.8%, 74.4% and 36,8% of the patients). The corresponding diagnostic yield for these test was 4.5%, 6.7%, 0% and 16.3%. Other procedures were (% of patients; diagnostic yield): Ct scan of the head (35.1%; 0%), transthoracic echocardiogram (24.8%; 6.9%), ECG monitoring (24.8%; 27.6%), EEG (19.7%; 0%), neurovascular imaging (19.7%; 0%), abdominal ultrasound (11.1%; 0%), Ct scan of the chest (11.1%; 23.1%), head up tilt test (7.7%; 11.1%), carotid sinus massage (3.4%; 0%), pulmonary ventilation perfusion scintigraphy (2.6%; 33%) and exercise test (1.7%; 0%). The most common cause of syncope was neurocardiogenic (58,5% of patients), followed by orthostatic (11.1%), cardiac (11.1%), unknown (9.3%), hyperventilation (3.4%), pulmonary embolism (2.5%), drug related (2.5%) and others (1.7%). CONCLUSION: Patients admitted in the ED for syncope undergo various investigations. However, the diagnostic yield is poor.The most common cause of syncope is neurocardiogenic, followed by orthostatic and cardiac. PMID- 21630608 TI - Do-not-resuscitate decisions in a large tertiary hospital: differences between wards and results of a hospital-wide intervention. AB - BACKGROUND: Despite the advent of palliative care, the quality of dying in the hospital remains poor. Differences in quality of end-of-life practice between hospital wards are well known in clinical practice but rarely have been investigated. METHODS: A prospective observation of do-not-resuscitate (DNR) decisions was conducted in patients dying in a Belgian university hospital before (115 patients) and after (113 patients) a hospital-wide intervention consisting in informing hospital staff on the law on Patient's Rights and the introduction of a new DNR form.The new DNR form puts more emphasis on the motivation of the DNR decision and on the description of the participants in the decision-making process. RESULTS: The completion of DNR forms improved after the intervention: physicians better documented who participated in DNR decisions (for participation of family: 63% after the intervention vs. 44% before the intervention, p = 0.022, for nurses: 27% vs. 14%, p = 0.047) and the motivation for these decisions (59% vs. 32%, p = 0.001). However, there was no difference in referral to the intensive care unit (ICU) at the end of life (in 40% of patients after and 37% before the intervention). Furthermore, the number of patients dying without DNR form on the wards was similar (13% and 8%). Surgical patients and patients with non-malignant diseases were more often referred to ICU at the end of life (71% in surgical vs. 35% in medical patients, p < 0.001 and 49% in patients with non malignant diseases vs. 23% in patients with malignancy, p < 0.001). Moreover, surgical patients less frequently received a DNR order (56% in surgical vs. 92% in medical patients, p = 0.007). CONCLUSIONS: The introduction of a new DNR form and informing hospital staff on patients' right to information did not improve physicians' end-of-life practice.Transition from life-prolonging treatment to a more palliative approach was less anticipated in surgical patients and patients with non-malignant diseases. PMID- 21630609 TI - Fluctuations of haemoglobinaemia in chronic haemodialysis patients. AB - In March 2008 and June 2009, an ad hoc working group of nephrologists discussed the status of anaemia therapy with erythropoiesis-stimulating agents [ESA] in patients on chronic haemodialysis, the phenomenon of fluctuations of haemoglobinaemia, and the need for individualisation of ESA treatment. The working group put together the following statements: (1) ESAs increase the haemoglobin concentration and adaptations of the ESA dose adjust the response according to a negative-feedback loop. The long lag time between an ESA dose change and its effect on erythropoiesis is cumbersome. The optimal haemoglobin target concentration is different for every haemodialysis patient; the lowest haemoglobin concentration upon which one could consistently demonstrate a positive subjective and objective clinical benefit in chronic dialysis is 11 g/dL, in contrast to the lowest haemoglobin concentration of 10 g/dL recommended in the current EMEA label for ESAs. (2) Intra-individual fluctuation of haemoglobinaemia over time is unavoidable, not only due to the ESA dose/haemoglobin response interaction, but also, and more importantly, due to the occurrence of acute illnesses and exacerbations of co-morbid conditions. Many different methodologies to characterise haemoglobin variability have been described but there is currently no universally applied definition of the phenomenon. (3) An impact of the haemoglobin level and the amplitude of the haemoglobin fluctuations on patient outcome has been observed. Without disclosing any causal relationship, worse outcomes were associated with haemoglobin fluctuations around the lower target level, but later on, more simply linked to the relative time spent below the haemoglobin concentration of 11 g/dL and to the administration of inappropriately high ESA doses in order to achieve the recommended haemoglobin target range. A plausible mechanism might be that acute illnesses blunt the patients' basal ESA sensitivity; this leads to subnormal and/or varying haemoglobin levels, currently initiating an ESA dose increase. The longer it takes the patient to recover from the acute illness, the more the prolongation of the clinically poor condition is to some extent maintained by the persistence of low haemoglobinaemia and/or by the administration of high ESA doses, and, as such, on their turn possibly contributing to an ultimate poor outcome. In the absence of clinical trials, recommendations should be offered how to proceed with the administration of ESAs as optimal as possible in periods of clinical instability. PMID- 21630610 TI - A woman with recurrent "infections" since birth--a new mevalonate kinase mutation. AB - A tired 32-year-old woman complaining of tiredness was referred for work-up of a possible immune deficiency. She had a history of recurrent infections since birth, which usually responded to antibiotics within a few days. Her mother, a nurse, had reported that early charts had disappeared. Munchausen's by proxy was suspected for years. Careful anamnesis indicated possible recurrent fever. Serum IgD levels were high, which led us to suspect Hyper IgD Syndrome. Sequencing of the mevalonate kinase gene revealed 2 mutations, leading to amino acid substitutions: one already described (V3771) and R40W: never reported before. Mevalonate kinase activity was very low in the patient's peripheral blood cells. We used the "Poly Phen" prediction program successfully. Our experiments confirmed the diagnosis of mevalonate kinase deficiency. We used steroids to abort recurrent crises. PMID- 21630611 TI - Polycythaemia vera presented with bilateral adrenal haemorrhage and adrenal insufficiency: a case report. AB - Adrenal insufficiency is rarely associated with adrenal haemorrhage. In this report, a case of polycythaemia vera with adrenal haemorrhage and insufficiency has been presented. PMID- 21630612 TI - Concomitant use of simvastatin and amiodarone resulting in severe rhabdomyolysis: a case report and review of the literature. AB - Myopathy, including rhabdomyolysis, is a well-known, albeit rare complication of statin therapy. Predisposing factors include comorbidities and the concomitant use of cytochrome P-450 (CYP) 3A4 inhibitors. We report a case of severe simvastatin-induced rhabdomyolysis triggered by the addition of amiodarone to previously well-tolerated chronic statin therapy. Physicians should be aware of the risk of this potentially severe drug interaction. The dose of simvastatin should be reduced (to 20 mg daily) when concomitant treatment with amiodarone is required, or preference should be given to pravastatin, rosuvastatin or fluvastatin, which are not metabolised by the CYP 3A4. PMID- 21630613 TI - Delayed diagnosis of abdominal mass due to morbid obesity. AB - Obesity has become an increasing problem in developed countries. Obesity is associated with many of the most common causes of morbidity and mortality, including diabetes mellitus, coronary artery diseases, sleep apnoea, and many types of cancers. Not only is morbid obesity associated to a greater extent to the many medical problems related to obesity, but the body habitus of a morbidly obese patient causes limitation to physical examination, radiologic evaluations, and therapeutic interventions that can delay diagnosis and treatment of the patient. We present a case of a morbidly obese patient whose diagnosis of liposarcoma was delayed for over a year as his obesity hindered proper diagnosis and medical treatment. PMID- 21630614 TI - Microscopic polyangiitis involving the breast. AB - We report a case of a 76-year-old woman, presenting with a persistent dry cough, subfebrility, arthralgia and myalgia, weight loss and a breast lesion. She has elevated inflammatory parameters, impaired renal function with proteinuria, bilateral lung nodules on computed tomography scan (CT scan) and a suspect lesion on mammography. A diagnosis of microscopic polyangiitis with involvement of the breast is made based on clinical and radiographic findings, with positive auto immune serology and histological confirmation. Although vasculitis of the breast is uncommon, this case illustrates that when a breast lesion is found, in combination with constitutional symptoms, we should think about the possibility of an anti-Neutrophil cytoplasmic antibodies (ANCA)-associated vasculitis. Therefore it may be important to perform auto-immune serology in these cases before proceeding to major surgery. PMID- 21630615 TI - Late vitamin K deficiency bleeding leading to a diagnosis of cystic fibrosis: a case report. AB - Vitamin K deficiency bleeding (VKDB) in infants still occurs despite worldwide use of prophylaxis. Clinical manifestations can be dramatic with over 50% of patients presenting with intracranial haemorrhage and a mortality rate of 20% in late vitamin K deficiency bleeding. Special attention should be given to infants with a high risk profile (preterm, breast feeding, cholestasis, malabsorption). A tentative diagnosis can be made observing quick normalisation of some easy-to perform haemostatic parameters (PT, aPTT) after administration of vitamin K. Nowadays, VKDB can still be the first clinical sign of diseases causing malabsorption of fat-soluble vitamins. In this case report, VKDB led to the diagnosis of cystic fibrosis, the most common fatal autosomal recessive disease among Caucasian people. PMID- 21630616 TI - Soft tissue infections with atypical mycobacteria in two patients with inflammatory rheumatic diseases using TNF-inhibitors and/or leflunomide. AB - Infections of the soft tissues due to atypical mycobacteria are relatively uncommon. We describe two cases. A 61-year-old woman with rheumatoid arthritis (RA) who was treated with the combination of the TNF inhibitor etanercept (Enbrel) and leflunomide (Arava), developed paronychia and cellulitis of the index finger due to Mycobacterium chelonae/abscessus complex. The patient was successfully treated with clarithromycin and surgical debridement. A second case describes a 50-year-old man with ankylosing spondylitis, receiving infliximab (Remicade) and low dose corticosteroids, who developed a granulomatous infection of the right thumb and forearm due to Mycobacterium marinum.The patient was successfully treated with clarithromycin and ethambutol. The increased risk for subcutaneous mycobacterial infections in these cases are probably related to the DMARD treatment with a TNF-inhibitor and leflunomide. PMID- 21630617 TI - Mediastinal extension of a goiter. PMID- 21630618 TI - Persistent secondary hyperparathyroidism. PMID- 21630619 TI - One guest on a visit in two hosts. PMID- 21630620 TI - A rare cause of severe bradycardia and hypotension: mad honey poisining. PMID- 21630621 TI - Engaging the next generation of health leaders: perspectives of emerging health leaders. AB - Identifying and engaging the next generation of emerging health leaders (EHLs) is a foremost challenge for the Canadian healthcare system. If intellectual capital is the currency of the 21st century, identifying and developing EHLs must be a strategic objective of innovative and progressive health organizations. We have integrated our experiential knowledge with some relevant findings from the literature and data collected from a cohort of EHLs to assist senior leaders and organizations with this challenge. PMID- 21630622 TI - When ordinary days become extraordinary: the response to H1N1 at the Izaak Walton Killam Health Centre. AB - In April 2009, the H1N1 swine flu virus entered Canada with a sentinel case presenting in a private school in Windsor, Nova Scotia, signaling the beginning of wave 1 of a pandemic. By June, the World Health Organization raised the alert level to 6. Wave 2 was expected in 4 to 6 months. Between waves, the IWK took the opportunity to revise pandemic plans that had been prepared for the H5N1 avian influenza virus to ensure that the organization would be prepared for the emergency response that would be required to manage large numbers of ill people seeking healthcare services for the pending second wave of the swine flu. PMID- 21630623 TI - Suburban cardiac screening: improving access to specialist services within a primary care network. AB - This article evaluates a cardiac screening program by analyzing wait times and exploring associations between administratively tracked variables and confirmed cardiac diagnosis. The findings indicate that the wait times for specialist consultation are shorter than previously reported in Alberta and age and sex have the strongest associations with a confirmed cardiac diagnosis. PMID- 21630624 TI - Towards sustainable traumatic brain injury care systems: healthcare leadership imperatives in Canada. AB - Traumatic brain injuries pose strategic population health challenges in the face of burgeoning clinical demands that continue to tax capital, financial, and social resource capacities. The sustainability of traumatic brain injury care systems depends on paradigmatic shifts in healthcare leadership thinking. In quest for high-performance care and sustained quality of life for traumatic brain injury patients, this article presents a unique paradigm of seven care performance layers and seven health leadership imperatives that together form the paradigm for the systemic sustainability of TBI care systems of the future. PMID- 21630625 TI - Ethical management in health services administration: a sequel. PMID- 21630626 TI - Understanding sleep-disordered breathing as a risk factor for hypertension and metabolic diseases: implications for clinical assessment. PMID- 21630627 TI - Factors affecting program completion in phase II cardiac rehabilitation. AB - BACKGROUND: Completion of a cardiac rehabilitation (CR) program post cardiac disease event promotes successful recovery and subsequent cardiovascular health. Attrition rates for CR programs have been reported as high as 65%. Little is known about the attrition population. PURPOSE: The purpose of this study was to describe demographic and clinical variables associated with non-completion of CR and to identify factors that led to attrition. METHODS: A comparative retrospective survey design was used to identify differences in demographic and clinical variables between patients who completed CR and those who did not. Prospectively, CR participants who dropped out received follow-up calls to identify reasons for program cessation. RESULTS: Demographic variables were not significantly different between the attrition group and the control group. Having a normal ECG during a pre-program stress test and having higher levels of pre program stress were significant for the attrition group. The most common reason for dropping out was physical health problems. Other influential factors included patients' perception that the exercise component of the program was too difficult and personal perceptions and reactions to the program. IMPLICATIONS: Patients entering CR who present in better physical risk categories with higher home or occupational stress levels may be at risk for dropping out. CR staff should monitor patients early for personal reactions to the program along with their response to physical exercise in order to address issues that promote program attrition. PMID- 21630628 TI - Patient-centred assessment of social support, health status and quality of life in patients with acute coronary syndrome. AB - BACKGROUND: Measurement of health status (HS) and social support are becoming increasingly accepted as tools to guide clinical decision-making and patient centred practice. PURPOSE: To assess self-reported HS, cardiac-health related quality of life and social support in subjects with a diagnosis of acute coronary syndrome (ACS). DESIGN: The study used a quantitative descriptive design. SAMPLE: 36 subjects with a diagnosis of ACS were selected from patients admitted to medical units at a teaching hospital in Toronto, Ontario. METHODS: One-time, semi structured interviews were conducted using valid and reliable cardiac-specific HS and social support measures. RESULTS: Analysis indicated that subjects with higher perceived social support and patients with higher income reported greater treatment satisfaction and C-HROL. Subjects with severe angina reported a higher perceived level of social support than those with more moderate physical limitation due to angina. CONCLUSION: Patients' social environment and HS significantly impact their satisfaction with treatment. Patient-centred measures assist in clinical decision-making, patient-centred care planning and patient involvement in their care. PMID- 21630629 TI - When blood runs cold: cold agglutinins and cardiac surgery. AB - Cold agglutinins are particular cold-reactive antibodies that react with red blood cells when the blood temperature drops below normal body temperature causing increased blood viscosity and red blood cell clumping. Most individuals with cold agglutinins are not aware of their presence, as these antibodies have little effect on daily living, often necessitating no treatment. However, when those with cold agglutinins are exposed to hypothermic situations or undergo procedures such as cardiopulmonary bypass with hypothermia during cardiac surgery, lethal complications of hemolysis, microvascular occlusion and organ failure can occur. By identifying those suspected of possessing cold agglutinins through a comprehensive nursing assessment and patient history, cold agglutinin screening can be performed prior to surgery to determine a diagnosis of cold agglutinin disease. With a confirmed diagnosis of cold agglutinin disease, the plan of care can be focused on measures to maintain the patient's blood temperature above the thermal amplitude throughout their hospitalization including the use of normothermic cardiopulmonary bypass with warm myocardial preservation techniques to prevent these fatal complications. Using a case report approach, the authors review the mechanism, clinical manifestations, detection and nursing management of a patient with cold agglutinins undergoing scheduled cardiac surgery. Cold agglutinin disease is rare. However, the risk to patients warrants an increased awareness of cold agglutinins and screening for those who are suspected of carrying these antibodies. PMID- 21630630 TI - My abstract was accepted--now what? A guide to effective conference presentations. AB - Although conference presentations are central to the dissemination of new nursing knowledge, public speaking can be a daunting prospect. The purpose of this article is to provide nurses with the essential tools to deliver a successful conference presentation. Accordingly, the all-too-familiar steps of assessment, planning, implementation, and evaluation provide an organizational framework for a practical guide to developing and executing a relatively stress-free, effective presentation. PMID- 21630631 TI - ...that exercise training is safe, useful and effective therapy and does not worsen cardiac function in heart failure? AB - Exercise training in heart failure is safe, useful and effective therapy in the management of heart failure patients. Contrary to the old belief, it does not worsen cardiac function. Smaller studies also reported reduction in mortality and morbidity associated with exercise training. The available data support training HF patients to improve exercise tolerance and symptoms. Patients should be properly reviewed by their physicians prior to engaging in exercise training. It is highly recommended that patients be referred to a cardiac rehabilitation program, especially those with severe or advanced heart failure. PMID- 21630633 TI - cis- and trans-.beta.-Sesquiphellandrol. Two new sesquiterpene alcohols from oil of ginger (Zingiber officinale). PMID- 21630632 TI - Self-assembly and photo-cross-linking of eight-armed PEG-PTMC star block copolymers. AB - Eight-armed poly(ethylene glycol)-poly(trimethylene carbonate) star block copolymers (PEG-(PTMC)(8)) linked by a carbamate group between the PEG core and the PTMC blocks were synthesized by the metal-free, HCl-catalyzed ring-opening polymerization of trimethylene carbonate using an amine-terminated eight-armed star PEG in dichloromethane. Although dye solubilization experiments, nuclear magnetic resonance spectroscopy, and dynamic light scattering clearly indicated the presence of aggregates in aqueous dispersions of the copolymers, no physical gelation was observed up to high concentrations. PEG-(PTMC(9))(8) was end-group functionalized using acryloyl chloride and photopolymerized in the presence of Irgacure 2959. When dilute aqueous dispersions of PEG-(PTMC(9))(8)-Acr were UV irradiated, chemically cross-linked PEG-PTMC nanoparticles were obtained, whereas irradiation of more concentrated PEG-(PTMC(9))(8)-Acr dispersions resulted in the formation of photo-cross-linked hydrogels. Their good mechanical properties and high stability against hydrolytic degradation make photo-cross-linked PEG-PTMC hydrogels interesting for biomedical applications such as matrices for tissue engineering and controlled drug delivery systems. PMID- 21630634 TI - Conic electrophoretic concentrator for charged macromolecules. AB - A simple, rapid, and highly effective technique for concentrating charged macromolecules is described which employs electrophoresis in a conic cell made of a dialysis membrane. The cell is partly submerged in electrolyte solution, and the level of solution slowly moves down during the process. The electric field within the cell is at its maximum in the area that is level with the surface of the external solution. This maximum value increases and its location moves downward following the decreasing level of external solution carrying downward and concentrating charged macromolecules. It has been demonstrated that proteins can be concentrated within 12-15 min by a factor of ~100,000 with the total yield of 60-80%. Concentrated proteins can be harvested from the nanoliter-sized cul-de sac of the conic concentrator using chemically activated magnetic beads. The presence of certain protein molecules linked to the bead's surface can be further revealed by specific reaction with a microarray of antibody molecules. Such "reversed magnetic array" format was applied to a cone-concentrated exhaled breath condensate (EBC) to reveal the presence of human immunoglobulin in the EBC and to estimate its concentration. The technique may be used for concentrating and detecting trace amounts of pathogens and toxins, in protein crystallization, and in many other applications. PMID- 21630635 TI - Single-cell metabolomics comes of age: new developments in mass spectrometry profiling and imaging. AB - Modern mass spectrometry is ready to explore individual cell metabolomes, and 4 massive advances in medicinal and biological applications may be expected in a near future. PMID- 21630636 TI - Polymeric complements to the Alzheimer's disease biomarker beta-amyloid isoforms Abeta1-40 and Abeta1-42 for blood serum analysis under denaturing conditions. AB - Treatment of Alzheimer's disease (AD) is plagued by a lack of practical and reliable methods allowing early diagnosis of the disease. We here demonstrate that robust receptors prepared by molecular imprinting successfully address current limitations of biologically derived receptors in displaying affinity for hydrophobic peptide biomarkers for AD under denaturing conditions. C-terminal epitope-imprinted polymers showing enhanced binding affinity for Abeta1-42 were first identified from a 96-polymer combinatorial library. This information was then used to synthesize molecularly imprinted polymers for both of the beta amyloid (Abeta) isoforms and a corresponding nonimprinted polymer. A solid-phase extraction method was developed to be compatible with sample loading under conditions of complete protein denaturation. This resulted in a method capable of quantitatively and selectively enriching a shorter C-terminal peptide corresponding to the sequences Abeta33-40 and Abeta33-42 as well as the full length sequence Abeta1-40 and Abeta1-42 from a 4 M guanidinum chloride solution. Application of the method to serum allowed selective, high-recovery extraction of both biomarkers at spiking levels marginally higher than clinically relevant concentrations found in cerebrospinal fluid. PMID- 21630637 TI - tert-Butylhydroquinone as a spectroscopic probe for the superoxide radical scavenging activity assay of biological samples. AB - As a more convenient and less costly alternative to electron spin resonance (ESR) and nonspecific nitroblue tetrazolium (NBT) and cytochrome c assays of superoxide radical (SR, O(2)(*-)) detection, a novel probe, tert-butylhydroquinone (TBHQ), is introduced for SR nonenzymatically generated in the phenazine methosulfate beta-nicotinamide adenine dinucleotide (PMS-NADH) system. SR attacks both TBHQ and SR scavengers incubated in solution for 30 min where scavengers compete with TBHQ for the O(2)(*-) produced. TBHQ, but not its O(2)(*-) oxidation product, tert-butyl-1,4-benzoquinone (TBBQ), is responsive to the CUPRAC (cupric reducing antioxidant capacity) spectrophotometric assay. The CUPRAC absorbance of the ethyl acetate extract of the incubation solution arising from the reduction of Cu(II)-neocuproine reagent by the remaining TBHQ was higher in the presence of O(2)(*-) scavengers (due to less conversion to TBBQ), the difference being correlated to the SR scavenging activity (SRSA) of the analytes. With the use of this reaction, a kinetic approach was adopted to assess the SRSA of amino acids, vitamins, and plasma and thiol antioxidants. This assay, applicable to small molecule antioxidants and tissue homogenates, proved to be efficient for cysteine, uric acid, and bilirubin, for which the widely used NBT test is nonresponsive. Thus, conventional problems of NBT assay arising from formazan insolubility and direct reduction of NBT by tested scavengers were overcome. PMID- 21630638 TI - Hispolon induces apoptosis and cell cycle arrest of human hepatocellular carcinoma Hep3B cells by modulating ERK phosphorylation. AB - Hispolon is an active phenolic compound of Phellinus igniarius , a mushroom that has recently been shown to have antioxidant, anti-inflammatory, and anticancer activities. This study investigated the antiproliferative effect of hispolon on human hepatocellular carcinoma Hep3B cells by using the MTT assay, DNA fragmentation, DAPI (4,6-diamidino-2-phenylindole dihydrochloride) staining, and flow cytometric analyses. Hispolon inhibited cellular growth of Hep3B cells in a time-dependent and dose-dependent manner, through the induction of cell cycle arrest at S phase measured using flow cytometric analysis and apoptotic cell death, as demonstrated by DNA laddering. Hispolon-induced S-phase arrest was associated with a marked decrease in the protein expression of cyclins A and E and cyclin-dependent kinase (CDK) 2, with concomitant induction of p21waf1/Cip1 and p27Kip1. Exposure of Hep3B cells to hispolon resulted in apoptosis as evidenced by caspase activation, PARP cleavage, and DNA fragmentation. Hispolon treatment also activated JNK, p38 MAPK, and ERK expression. Inhibitors of ERK (PB98095), but not those of JNK (SP600125) and p38 MAPK (SB203580), suppressed hispolon-induced S-phase arrest and apoptosis in Hep3B cells. These findings establish a mechanistic link between the MAPK pathway and hispolon-induced cell cycle arrest and apoptosis in Hep3B cells. PMID- 21630639 TI - Boron-assisted transformation to rod-like graphitic carbons from multi-walled carbon nanotubes in boron-mixed multi-walled carbon nanotube solids. AB - We produced boron-mixed multi-walled carbon nanotube solids (B-mixed MWCNT solids) by heating and pressing the powder of purified MWCNTs mixed with 1, 5, and 10 wt % boron in the temperature range 1400-1800 degrees C every 200 degrees C under a constant pressure of 20 MPa in vacuo, and investigated the influence of boron addition on nanotube structure and the mechanical and electrical properties of the resulting B-mixed MWCNT solids. The structure of the prepared material was characterized by scanning electron microscopy equipped with energy-dispersive X-ray spectroscopy, high-resolution transmission electron microscopy-electron energy loss spectroscopy, Raman scattering spectroscopy, and X-ray diffraction, and their mechanical properties and conductivity were measured using a mechanical and Vickers indentation tester and an electric resistor, respectively. It is notable that part of the nanotubes in the B-mixed MWCNT solids solidified at 1800 degrees C had dramatically changed into rod-like graphitic carbons (RLGCs). The occupancy distribution of RLGCs increased with increasing boron contents. However, boron was not detected in the energy-loss near-edge structure spectrum of RLGCs. Furthermore, RLGCs were not observed in the boron-unmixed sample treated with the same solidified condition, indicating that adding boron causes a remarkable ability to transform the phase of MWCNT. Transformation from MWCNTs to RLGCs resulted in increased specific bending strength and modulus, Vickers hardness, and electrical conductivity of B-mixed MWCNT solids with increasing boron content and solidified temperature. PMID- 21630640 TI - PPII propensity of multiple-guest amino acids in a proline-rich environment. AB - There has been considerable debate about the intrinsic PPII propensity of amino acid residues in denatured polypeptides. Experimentally, this scale is based on the behavior of guest amino acid residues placed in the middle of proline-based hosts. We have used classical molecular dynamics simulations combined with replica-exchange methods to carry out a comprehensive analysis of the conformational equilibria of proline-based host oligopeptides with multiple guest amino acids including alanine, glutamine, valine, and asparagine. The tracked structural characteristics include the secondary structural motifs based on the Ramachandran angles and the cis/trans isomerization of the prolyl bonds. In agreement with our recent study of single amino acid guests, we did not observe an intrinsic PPII propensity in any of the guest amino acids in a multiple-guest setting. Instead, the experimental results can be explained in terms of (i) the steric restrictions imposed on the C-terminal guest amino acid that is immediately followed by a proline residue and (ii) an increase in the trans content of the prolyl bonds due to the presence of guest residues. In terms of the latter, we found that the more guests added to the system, the larger the increase in the trans content of the prolyl bonds, which results in an effective increase in the PPII content of the peptide. PMID- 21630641 TI - Apoptosis goes on a chip: advances in the microfluidic analysis of programmed cell death. AB - Recent years have brought enormous progress in cell-based lab-on-a-chip technologies, allowing dynamic studies of cell death with an unprecedented accuracy. As interest in the microfabricated technologies for cell-based bioassays is rapidly gaining momentum, we highlight the most promising technologies that provide a new outlook for the rapid assessment of programmed and accidental cell death and are applicable in drug discovery, high-content drug screening, and personalized clinical diagnostics. PMID- 21630642 TI - Metal binding ability of cysteine-rich peptide domain of ZIP13 Zn2+ ions transporter. AB - The coordination modes and thermodynamic stabilities of the complexes of the cysteine-rich N-terminal domain fragment of the ZIP13 zinc transporter (MPGCPCPGCG-NH(2)) with Zn(2+), Cd(2+), Bi(3+), and Ni(2+) have been studied by potentiometric, mass spectrometric, NMR, CD, and UV-vis spectroscopic methods. All of the studied metals had similar binding modes, with the three thiol sulfurs of cysteine residues involved in metal ion coordination. The stability of the complexes formed in solution changes in the series Bi(3+) ? Cd(2+) > Zn(2+) > Ni(2+), the strongest being for bismuth and the weakest for nickel. The N terminal fragment of the human metalothionein-3 (MDPETCPCP-NH(2)) and unique histidine- and cysteine-rich domain of the C-terminus of Helicobacter pyroli HspA protein (Ac-ACCHDHKKH-NH(2)) have been chosen for the comparison studies. It confirmed indirectly which groups were the anchoring ones of ZIP13 domain. Experimental data from all of the used techniques and comparisons allowed us to propose possible coordination modes for all of the studied ZIP13 complexes. PMID- 21630643 TI - Synthesis, crystal structures, and magnetic properties of a new family of heterometallic cyanide-bridged Fe(III)2M(II)2 (M=Mn, Ni, and Co) square complexes. AB - New heterobimetallic tetranuclear complexes of formula [Fe(III){B(pz)(4)}(CN)(2)(MU-CN)Mn(II)(bpy)(2)](2)(ClO(4))(2).CH(3)CN (1), [Fe(III){HB(pz)(3)}(CN)(2)(MU-CN)Ni(II)(dmphen)(2)](2)(ClO(4))(2).2CH(3)OH (2a), [Fe(III){B(pz)(4)}(CN)(2)(MU-CN)Ni(II)(dmphen)(2)](2)(ClO(4))(2).2CH(3)OH (2b), [Fe(III){HB(pz)(3)}(CN)(2)(MU-CN)Co(II)(dmphen)(2)](2)(ClO(4))(2).2CH(3)OH (3a), and [Fe(III){B(pz)(4)}(CN)(2)(MU-CN)Co(II)(dmphen)(2)](2)(ClO(4))(2).2CH(3)OH (3b), [HB(pz)(3)(-) = hydrotris(1-pyrazolyl)borate, B(Pz)(4)(-) = tetrakis(1 pyrazolyl)borate, dmphen = 2,9-dimethyl-1,10-phenanthroline, bpy = 2,2' bipyridine] have been synthesized and structurally and magnetically characterized. Complexes 1-3b have been prepared by following a rational route based on the self-assembly of the tricyanometalate precursor fac [Fe(III)(L)(CN)(3)](-) (L = tridentate anionic ligand) and cationic preformed complexes [M(II)(L')(2)(H(2)O)(2)](2+) (L' = bidentate alpha-diimine type ligand), this last species having four blocked coordination sites and two labile ones located in cis positions. The structures of 1-3b consist of cationic tetranuclear Fe(III)(2)M(II)(2) square complexes [M = Mn (1), Ni (2a and 2b), Co (3a and 3b)] where corners are defined by the metal ions and the edges by the Fe CN-M units. The charge is balanced by free perchlorate anions. The [Fe(L)(CN)(3)](-) complex in 1-3b acts as a ligand through two cyanide groups toward two divalent metal complexes. The magnetic properties of 1-3b have been investigated in the temperature range 2-300 K. A moderately strong antiferromagnetic interaction between the low-spin Fe(III) (S = 1/2) and high spin Mn(II) (S = 5/2) ions has been found for 1 leading to an S = 4 ground state (J(1) = -6.2 and J(2) = -2.7 cm(-1)), whereas a moderately strong ferromagnetic interaction between the low-spin Fe(III) (S = 1/2) and high-spin Ni(II) (S = 1) and Co(II) (S = 3/2) ions has been found for complexes 2a-3b with S = 3 (2a and 2b) and S = 4 (3a and 3b) ground spin states [J(1) = +21.4 cm(-1) and J(2) = +19.4 cm(-1) (2a); J(1) = +17.0 cm(-1) and J(2) = +12.5 cm(-1) (2b); J(1) = +5.4 cm(-1) and J(2) = +11.1 cm(-1) (3a); J(1) = +8.1 cm(-1) and J(2) = +11.0 cm(-1) (3b)] [the exchange Hamiltonian being of the type H = -J(S(i).S(j))]. Density functional theory (DFT) calculations have been used to substantiate the nature and magnitude of the exchange magnetic coupling observed in 1-3b and also to analyze the dependence of the exchange magnetic coupling on the structural parameters of the Fe-C-N-M skeleton. PMID- 21630644 TI - Human hemolysate glycated proteome. AB - Despite continuous advances in hyperglycemia treatments, a precise control through monitoring of glucose and glycated hemoglobin remains in most diabetic patients as the diagnosis/prognosis tool. An alternative perspective could be the discovery and quantitation of new blood glycated proteins formed by nonenzymatic reaction with circulatory glucose. As a result, the human hemolysate is an incomparable source of glycated proteins to further monitor glycemia and interpret changes at the level of this post-translational modification. The human hemolysate is here studied based on the differential labeling of proteins with isotopically labeled-glucose ([(13)C(6)] glucose), named glycation isotopic labeling. Due to the chemoselectivity of glycation, only preferential targets are labeled by this protocol. The approach provides qualitative data through the detection of preferential protein glycation sites as well as quantitative information to evaluate the abundance of this modification. This strategy was applied to human hemolysate samples corresponding to different glycemic states estimated by laboratory-certified concentrations of glycated hemoglobin. The glycation level of each protein can then be employed to interpret the effect of glucose exposition as a consequence of glycemic unbalance. This information should provide new molecular insights into protein glycation mechanisms that might generate a new hypothesis to clinicians to improve the understanding of underlying pathologies associated to prolonged hyperglycemia. PMID- 21630645 TI - Exploring matrix effects and quantification performance in metabolomics experiments using artificial biological gradients. AB - Metabolomics has become an integral part of many life-science applications but is technically still very challenging. Numerous analytical approaches are needed as metabolites have very broad concentration ranges and extremely diverse chemical properties. Configuring a metabolomics pipeline and exploring its merits is a complex task that depends on effective and transparent evaluation procedures. Unfortunately, there are no widely applicable methods to evaluate how well acquired data can approximate actual concentration differences. Here, we introduce a powerful approach that provides semiquantitative calibration curves over a biologically defined concentration range for all detected compounds. By performing metabolomics on a stepwise gradient between two biological specimens, we obtain a data set where each peak would ideally show a linear dependency on the mixture ratio. An example gradient between extracts of tomato leaf and fruit demonstrates good calibration statistics for a large proportion of the peaks but also highlights cases with strong background-dependent signal interference. Analysis of artificial biological gradients is a general and inexpensive tool for calibration that greatly facilitates data interpretation, quality control and method comparisons. PMID- 21630646 TI - Ruthenium hydride complexes of the hindered phosphine ligand tris(3 diisopropylphosphinopropyl)phosphine. AB - The synthesis and characterization of the novel hindered tripodal phosphine ligand P(CH(2)CH(2)CH(2)P(i)Pr(2))(3) (P(3)P(3)(iPr)) (1) are reported, along with the synthesis and characterization of ruthenium chloro and hydrido complexes of 1. Complexes [RuCl(P(3)P(3)(i)Pr)][BPh(4)] (2[BPh(4)]), RuH(2)(P(3)P(3)(i)Pr) (3), and [Ru(H(2))(H)(P(3)P(3)(iPr))][BPh(4)] (4[BPh(4)]) were characterized by crystallography. Complex 2 is fluxional in solution, and low-temperature NMR spectroscopy of the complex correlates well with two dynamic processes, an exchange between stereoisomers and a faster turnstile-type exchange within one of the stereoisomers. PMID- 21630647 TI - Excited-state intermolecular proton transfer of firefly luciferin V. Direct proton transfer to fluoride and other mild bases. AB - We studied the direct proton transfer (PT) from electronically excited D luciferin to several mild bases. The fluorescence up-conversion technique is used to measure the rise and decay of the fluorescence signals of the protonated and deprotonated species of D-luciferin. From a base concentration of 0.25 M or higher the proton transfer rates to the fluoride, dihdyrogen phosphate or acetate bases are fast and comparable. The fluorescence signals are nonexponential and complex. We suggest that the fastest decay component arises from a direct proton transfer process from the hydroxyl group of D-luciferin to the mild base. The proton donor and acceptor molecules form an ion pair prior to photoexcitation. Upon photoexcitation solvent rearrangement occurs on a 1 ps time-scale. The PT reaction time constant is ~2 ps for all three bases. A second decay component of about 10 ps is attributed to the proton transfer in a contact pair bridged by one water molecule. The longest decay component is due to both the excited-state proton transfer (ESPT) to the solvent and the diffusion-assisted PT process between a photoacid and a base pair positioned remotely from each other prior to photoexcitation. PMID- 21630648 TI - Cholesteric liquid crystals doped with dodecylamine for detecting aldehyde vapors. AB - In this paper, we report a study of using cholesteric liquid crystals (CLCs) doped with dodecylamine for detecting aldehyde vapors. CLCs doped with dodecylamine show color change from red to yellow-green upon exposure to 300 ppmv pentyl aldehyde within 60 s. In contrast, no colorimetric response is observed when pure CLCs are used. Characterization by using FT-IR shows that the possible mechanism responsible for the colorimetric response is the formation of an imine bond between dodecylamine and pentyl aldehyde. A new C=N peak at 1670 cm(-1) appears in the spectrum after the exposure to aldehyde vapors. The CLCs doped with dodecylamine show good selectivity for pentyl aldehyde; they do not respond to 200 ppmv pentyl alcohol, pentylamine, acetone, ethanol, and water vapor. This study demonstrates the potential applications of doped CLCs as low-cost and portable gas sensors. PMID- 21630649 TI - Formation of supported lipid bilayers at surfaces with controlled curvatures: influence of lipid charge. AB - We have developed and characterized novel biomimetic membranes, formed at nanostructured sensor substrates with controlled curvatures, motivated by the many biological processes that involve membrane curvature. Model systems with convex nanostructures, with radii of curvatures (ROCs) of 70, 75, and 95 nm, were fabricated utilizing colloidal assembly and used as substrates for supported lipid bilayers (SLBs). The SLBs were formed via vesicle adsorption and rupture, and the vesicle deposition pathway was studied by means of quartz crystal microbalance with dissipation (QCM-D) and fluorescence microscopy. SLBs conforming to the underlying nanostructured surfaces, which exhibit increased surface area with decreased ROC, were confirmed from excess mass, monitored by QCM-D, and excess total fluorescence intensities. The formation of SLBs at the nanostructured surfaces was possible, however, depending on the ROC of the structures and the lipid vesicle charge the quality varied. The presence of nanostructures was shown to impair vesicle rupture and SLB formation was progressively hindered at surfaces with structures of decreasing ROCs. The introduction of a fraction of the positively charged lipid POEPC in the lipid vesicle membrane allowed for good quality and conformal bilayers at all surfaces. Alternatively, for vesicles formed from lipid mixtures with a fraction of the negatively charged lipid POPS, SLB formation was not at all possible at surfaces with the lowest ROC. Interestingly, the vesicle adsorption rate and the SLB formation were faster at surfaces with nanostructures of progressively smaller ROCs at high ratios of POPS in the vesicles. Development of templated SLBs with controlled curvatures provides a new experimental platform, especially at the nanoscale, at which membrane events such as lipid sorting, phase separation, and protein binding can be studied. PMID- 21630650 TI - Effects of aggregation on the excitation dynamics of LH2 from Thermochromatium tepidum in aqueous phase and in chromatophores. AB - We carried out femtosecond magic-angle and polarized pump-probe spectroscopies for the light-harvesting complex 2 (LH2) from Thermochromatium (Tch.) tepidum in aqueous phase and in chromatophores. To examine the effects of LH2 aggregation on the dynamics of excitation energy transfer, dominant monodispersed and aggregated LH2s were prepared by controlling the surfactant concentrations. The aqueous preparations solubilized with different concentrations of n-dodecyl-beta-D maltoside (DDM) show similar visible-to-near-infrared absorption spectra, but distinctively different aggregation states, as revealed by using dynamic light scattering. The B800 -> B850 intra-LH2 energy transfer time was determined to be 1.3 ps for isolated LH2, which, upon aggregation in aqueous phase or clustering in chromatophores, shortened to 1.1 or 0.9 ps, respectively. The light-harvesting complex 1 (LH1) of this thermophilic purple sulfur bacterium contains bacteriochlorophyll a absorbing at 915 nm (B915), and the LH2(B850) -> LH1(B915) intercomplex transfer time in chromatophores was found to be 6.6 ps. For chromatophores, a depolarization time of 21 ps was derived from the anisotropy kinetics of B850*, which is attributed to the migration of B850* excitation before being trapped by LH1. In addition, the B850* annihilation is accelerated upon LH2 aggregation in aqueous phase, but it is much less severe upon LH2 clustering in the intracytoplasmic membrane. These results are helpful in understanding the light-harvesting function of a bacterial photosynthetic membrane incorporating different types of antenna complexes. PMID- 21630651 TI - Electric-field-induced perfect anti-nematic order in isotropic aqueous suspensions of a natural beidellite clay. AB - We study the electric-field-induced birefringence and orientational order in the isotropic phase of aqueous suspensions of exfoliated natural beidellite clay particles, thin (L = 0.65 nm) flat charged sheets with high aspect ratio, D/L ~ 300. Our electric birefringence experiment is optimized for aqueous suspensions of colloidal particles, with a high frequency a.c. electric field, nu ~ 1 MHz, applied by two external electrodes to a thin flat sample, sealed in an optical capillary. In isotropic and biphasic samples, we observed strong field-induced birefringence Deltan(E), saturating at moderate E(sat) field to a plateau Deltan(sat) proportional to the volume fraction phi. The field-induced order parameter S(E) is negative and saturates to S(sat) = -0.5 above E(sat). This corresponds to a perfect "anti-nematic" order, i.e. the normals of the beidellite particles are perpendicular to the field, without any preferred azimuthal direction. The measured specific excess polarizability DeltaA(sp) is among the highest data reported for other strongly anisometric dielectric and metal particles. We explain the high DeltaA(sp) value with the strong induced polarization of the electric double layer of counterions at the charged particle/electrolyte interface. The estimated equivalent conductivity of the beidellite particle K(eq) = 2 K(sigma)/L is several orders of magnitude larger than the bulk conductivity of the electrolyte K(e), resulting in a metal-like behavior of the beidellite disks under field. In the isotropic regions of biphasic nematic/isotropic samples, the excess polarizability is further enhanced by an order of magnitude, indicating collective reorientation of the particles. We propose that this enhancement might be due to pretransitional fluctuations of the spontaneous nematic order S(N) of the colloidal suspension and/or formation of chains of particles, with antinematic order of the beidellite disks in the chains. PMID- 21630652 TI - Influence of pyrogenic particles on the micromechanical behavior of thin sol-gel layers. AB - Coatings based on sol-gel technology with different types of nanoparticles embedded into the sol-gel matrix were fabricated, and the resulting properties were investigated. Pyrogenic silica nanoparticles were added to the sol before coating. The silica particles varied in primary particle size and agglomerate size, and in their surface modification. The particles were wetted in ethanol and dispersed to certain finenesses. The difference in agglomerate size was partly caused by varying particle types, but also by the dispersing processes that were applied to the particles. The resulting coatings were examined by visual appearance and SEM microscopy. Furthermore, their micromechanical properties were determined by nanoindentation. The results show an important influence from the added nanoparticles and their properties on the visual appearance as well as the micromechanical behavior of the sol-gel coatings. It is shown that, in fact, the particle size distribution can have a major impact on the coating properties as well as the surface modification. PMID- 21630653 TI - Antimicrobial activity of olive solutions from stored Alpeorujo against plant pathogenic microorganisms. AB - The aim of this work was to assess the in vitro antimicrobial effects that wastewaters from alpeorujo oil extraction have against phytopathogenic bacteria and fungi. Alpeorujo was stored for 6 months and then processed to extract its oil, pomace, and a new liquid waste (OWSA), which was characterized by its content in phenolic compounds. OWSA at 20% decreased bu >4 log the population of Erwinia spp., Pseudomonas spp., and Clavibacter spp. viable cells in test tubes, whereas OWSA at 50% in agar medium was necessary to inhibit mycelial growth of most fungi. It was found that the bactericidal effect was due to the joint action of low molecular mass phenolic compounds, although neither hydroxytyrosol, its glucosides, hydroxytyrosol glycol, nor a glutaraldehyde-like compound individually explained this bioactivity. Hence, OWSA constitutes a promising natural solution to fight plant phytopathogenic bacteria and fungi. PMID- 21630654 TI - Metal interactions at the biochar-water interface: energetics and structure sorption relationships elucidated by flow adsorption microcalorimetry. AB - Plant-derived biochars exhibit large physicochemical heterogeneity due to variations in biomass chemistry and combustion conditions. However, the influence of biochar heterogeneity on biochar-metal interaction mechanisms has not been systematically described. We used flow adsorption microcalorimetry to study structure-sorption relationships between twelve plant-derived biochars and two metals (K(+) and Cd(2+)) of different Lewis acidity. Irrespective of the biochar structure, sorption of K(+) (a hard Lewis acid) occurred predominantly on deprotonated functional groups via ion exchange with molar heats of adsorption (DeltaH(ads)) of -4 kJ mol(-1) to -8 kJ mol(-1). By comparison, although ion exchange could not be completely ruled out, our data pointed to Cd(2+) (a soft Lewis acid) sorption occurring predominantly via two distinct cation-pi bonding mechanisms, each with DeltaH(ads) of +17 kJ mol(-1). The first, evident in low charge-low carbonized biochars, suggested Cd(2+)-pi bonding to soft ligands such as -C ? O; while the second, evident in low charge-highly carbonized biochars, pointed to Cd(2+)-pi bonding with electron-rich domains on aromatic structures. Quantitative contributions of these mechanisms to Cd(2+) sorption can exceed 3 times that expected for ion exchange and therefore could have significant implications for the biogeochemical cycling of metals in fire-impacted or biochar amended systems. PMID- 21630655 TI - Drop coating deposition Raman (DCDR) for microcystin-LR identification and quantitation. AB - A drop coating deposition Raman (DCDR) method was developed for the analysis of 2 200 ng samples of microcystin-LR (MC-LR), a ubiquitous and deadly hepatotoxin secreted by cyanobacteria. Solid phase extraction (SPE) of the toxin from a water sample enabled identification of MC-LR at 5 MUg/L to 100 mg/L concentrations, and the collected results suggest lower detection limits can be readily attained following DCDR substrate modification. The DCDR process was applied to aqueous sample volumes of 0.5-20 MUL that generated sample deposits from which MC-LR Raman spectra could be obtained within seconds. Larger volume samples were not required to improve spectral resolution. Volumes of 2 MUL were ideal, producing "coffee-ring" MC-LR deposits that displayed distinct MC-LR Raman signals with high signal-to-noise within 1 s for a 200 ng sample and 300 s for a 2 ng sample. A linear correlation between Raman signal intensity and concentration was observed for 2-100 ng MC-LR samples after signal normalization. Reproducible MC LR Raman spectra were collected from both fresh and aged samples. The presence of dissolved organic matter (DOM) did not preclude MC-LR identification in DCDR deposits of 3 MUg of DOM mixed with 0.7 MUg of MC-LR. Application of DCDR to environmental samples will require sample purification such as SPE before analysis, including critical cartridge wash and toxin rinsing steps. Raman based methods may one day facilitate simpler and faster sample throughput than traditional MC-LR detection methods. PMID- 21630656 TI - Ultrafast intersystem crossing and spin dynamics of zinc meso tetraphenylporphyrin covalently bound to stable radicals. AB - tert-Butylphenylnitroxide (BPNO(*)) and alpha,gamma-bisdiphenylene-beta phenylallyl (BDPA(*)) stable radicals are each attached to zinc meso tetraphenylporphyrin (ZnTPP) at a fixed distance using one of the ZnTPP phenyl groups. BPNO(*) and BDPA(*) are oriented para (1 and 3, respectively) or meta (2 and 4, respectively) relative to the porphyrin macrocycle. Following photoexcitation of 1-4, transient optical absorption spectroscopy is used to observe excited state quenching of (1)*ZnTPP by the radicals and time-resolved electron paramagnetic resonance (TREPR) spectroscopy is used to monitor the spin dynamics of the paramagnetic product states. The presence of BPNO(*) or BDPA(*) accelerates the intersystem crossing rate of (1)*ZnTPP about 10- to 500-fold in 1 4 depending on the structure compared to that of (1)*ZnTPP itself. In addition, the lifetime of (3)*ZnTPP in 1 is shorter than that of (3)*ZnTPP itself as a result of enhanced intersystem crossing (EISC) from (3)*ZnTPP to the ground state. The TREPR spectra of the three unpaired spins produced within 1 and 2 show spin-polarized excited doublet (D(1)) and quartet (Q) states and subsequent formation of a spin-polarized ground state radical (D(0)). All three signals are absorptive for 1 and emissive for 2. Polarization inversion of the Q state is observed on a tens of nanoseconds time scale in 2, while no polarization inversion is observed for 1. The lack of polarization inversion in 1 is attributed to the short lifetime of the doublet-quartet manifold as a result of the very large exchange interaction. The TREPR spectra of 3 and 4 show ground state radical polarization at X-band (9.5 GHz) at room temperature, but not at 85 K, and similarly no polarization is observed at W-band (94 GHz). No evidence of excited doublet or quartet states is observed, indicating that the exchange interaction is both weak and temperature dependent. These results show that although ultrafast EISC produces (3)*ZnTPP within 1-4, the magnitude of the exchange interactions between the three relevant spins in the resulting (3)*ZnTPP BPNO(*) and (3)*ZnTPP-BDPA(*) systems dramatically alters their spin dynamics. PMID- 21630657 TI - One-pot tuning of Au nucleation and growth: from nanoclusters to nanoparticles. AB - We describe a simple and effective method to obtain colloidal surface functionalized Au nanoparticles. The method is primarily based on irradiation of a gold solution with high-flux X-rays from a synchrotron source in the presence of 11-mercaptoundecanoic acid (MUA). Extensive tests of the products demonstrated high colloidal density as well as excellent stability, shelf life, and biocompatibility. Specific tests with X-ray diffraction, UV-visible spectrometry, visible microscopy, Fourier transform infrared spectroscopy, dark-field visible light scattering microscopy, and transmission electron microscopy demonstrated that MUA, being an effective surfactant, not only allows tunable size control of the nanoparticles, but also facilitates functionalization. The nanoparticle sizes were 6.45 +/- 1.58, 1.83 +/- 1.21, 1.52 +/- 0.37 and 1.18 +/- 0.26 nm with no MUA and with MUA-to-Au ratios of 1:2, 1:1, and 3:1. The MUA additionally enabled functionalization with l-glycine. We thus demonstrated flexibility in controlling the nanoparticle size over a large range with narrow size distribution. PMID- 21630658 TI - Structural and electronic characterization of non-heme Fe(II)-nitrosyls as biomimetic models of the Fe(B) center of bacterial nitric oxide reductase. AB - The detoxification of nitric oxide (NO) by bacterial NO reductase (NorBC) has gained much attention as this reaction provides a paradigm as to how NO can be detoxified anaerobically in cells. However, a clear mechanistic picture of how the heme/non-heme active site of NorBC activates NO is lacking, mostly as a result of insufficient knowledge about the properties of the non-heme iron(II)-NO adduct. Here we report the first biomimetic model complexes for this species that closely resemble the coordination environment found in the protein, using the ligands BMPA-Pr and TPA. The systematic investigation of these compounds allowed us to gain key insight into the electronic structure and geometric properties of high-spin non-heme iron(II)-NO adducts. In particular, we show how small changes in the ligand environment of iron could be used by NorBC to greatly modulate the properties, and hence, the reactivity of this species. PMID- 21630659 TI - Anticancer activities of thelephantin O and vialinin A isolated from Thelephora aurantiotincta. AB - Thelephora aurantiotincta is an edible mushroom belonging to the genus Thelephora; it grows in symbiosis with pine trees. Recently, phytochemical investigations have revealed that the genus Thelephora is an abundant source of p terphenyl derivatives. However, their bioactivity has not yet been well characterized. In screening for natural materials with anticancer activity, a T. aurantiotincta ethanol extract (TAE) was found to decrease cell viability in human hepatocellular carcinoma cells (HepG2). In this study, a new p-terphenyl derivative, thelephantin O, and a known compound, vialinin A, were isolated as the principal bioactive components of TAE. These compounds decreased cell viability in HepG2 and human colonic carcinoma cells (Caco2), but not in noncancerous human hepatocytes. This is the first report of the isolation from T. aurantiotincta of selective cytotoxic agents against cancer cells. PMID- 21630660 TI - Fish proteins as targets of ferrous-catalyzed oxidation: identification of protein carbonyls by fluorescent labeling on two-dimensional gels and MALDI TOF/TOF mass spectrometry. AB - Protein oxidation in fish meat is considered to affect negatively the muscle texture. An important source of free radicals taking part in this process is Fenton's reaction dependent on ferrous ions present in the tissue. The aim of this study was to investigate the susceptibility of cod muscle proteins in sarcoplasmic and myofibril fractions to in vitro metal-catalyzed oxidation and to point out protein candidates that might play a major role in the deterioration of fish quality. Extracted control proteins and proteins subjected to free radicals generated by Fe(II)/ascorbate mixture were labeled with fluorescein-5 thiosemicarbazide (FTSC) to tag carbonyl groups and separated by two-dimensional gel electrophoresis. Consecutive visualization of protein carbonyl levels by capturing the FTSC signal and total protein levels by capturing the SyproRuby staining signal allowed us to quantify the relative change in protein carbonyl levels corrected for changes in protein content. Proteins were identified using MALDI-TOF/TOF mass spectrometry and homology-based searches. The results show that freshly extracted cod muscle proteins exhibit a detectable carbonylation background and that the incubation with Fe(II)/ascorbate triggers a further oxidation of both sarcoplasmic and myofibril proteins. Different proteins exhibited various degrees of sensitivity to oxidation processes. Glyceraldehyde 3 phosphate dehydrogenase (GAPDH), nucleoside diphosphate kinase B (NDK), triosephosphate isomerase, phosphoglycerate mutase, lactate dehydrogenase, creatine kinase, and enolase were the sarcoplasmic proteins most vulnerable to ferrous-catalyzed oxidation. Moreover, NDK, phosphoglycerate mutase, and GAPDH were identified in several spots differing by their pI, and those forms showed different susceptibilities to metal-catalyzed oxidation, indicating that post translational modifications may change the resistance of proteins to oxidative damage. The Fe(II)/ascorbate treatment significantly increased carbonylation of important structural proteins in fish muscle, mainly actin and myosin, and degradation products of those proteins were observed, some of them exhibiting increased carbonylation levels. PMID- 21630661 TI - Enzymatic synthesis of lipophilic rutin and vanillyl esters from fish byproducts. AB - Lipase-catalyzed synthesis of lipophilic phenolic antioxidants was carried out with a concentrate of n-3 polyunsaturated fatty acids (PUFAs), recovered from oil extracted from salmon ( Salmon salar ) byproduct. Vanillyl alcohol and rutin were selected for the esterification reaction, and obtained esters yields were 60 and 30%, respectively. The antioxidant activities of the esters were compared with those of commercial butylated hydroxytoluene (BHT) and alpha-tocopherol using DPPH radical scavenging and thiobarbituric acid assays. In the DPPH assay, rutin esters showed better activity than vanillyl esters, and on the contrary in lipophilic medium, vanillyl esters were found to be superior to rutin esters. In bulk oil system, the antioxidant activities of rutin and vanillyl derivatives were lower than that of BHT and alpha-tocopherol, but in emulsion, they showed better activity than alpha-tocopherol. By attaching to natural phenolics, the PUFAs are protected against oxidation, and PUFA improves the hydrophobicity of the phenolic, which could enhance its function in lipid systems. PMID- 21630662 TI - A perspective for biowaivers of human bioequivalence studies on the basis of the combination of the ratio of AUC to the dose and the biopharmaceutics classification system. AB - The ratio of AUC to the dose (AUC/dose) was previously found as a parameter that predicts a risk of bioinequivalence of oral drug products. On the basis of the combination of this parameter and the biopharmaceutics classification system (BCS), a perspective for biowaivers of human bioequivalence studies is discussed. Databases of bioequivalence studies using immediate-release solid oral dosage forms were disclosed by 6 Japanese generic pharmaceutical companies, and the number of subjects required for demonstrating bioequivalence between generic and reference products was plotted as a function of AUC/dose for each BCS category. A small variation in the number of subjects was constantly observed in bioequivalence studies using dosage forms containing an identical BCS class 1 or class 3 drug, even though formulations of the generic product differ between companies. The variation was extremely enlarged when the drugs were substituted with BCS class 2 drugs. Rate-determining steps in oral absorption of highly water soluble BCS class 1 and class 3 drugs are independent of formulations when there is no significant difference in the in vitro dissolution profiles between formulations. The small variation observed for both BCS categories indicates that the number of subjects converges into one value for each drug. Our analysis indicates the appropriateness of biowaiver of bioequivalence studies for immediate-release solid oral dosage forms containing not only BCS class 1 drugs but also class 3 drugs. PMID- 21630664 TI - Laser-induced charge separation in CdSe nanowires. AB - A combination of electrostatic force microscopy and optical microscopy was used to investigate the charge state of individual CdSe nanowires upon local illumination with a focused laser beam. The nanowires were found to be positively charged at the excitation spot and negatively charged at the distant end(s). For high laser powers, the amount of accumulated charges increases logarithmically with the laser power. These effects are described by a diffusion-based model where the results are in good agreement with the experimentally observed effects. On the basis of this model the charge imbalance along the nanowire should establish in the course of nanoseconds. The net charge separation within homogeneous nanowires upon local illumination is of importance for several electronic devices. PMID- 21630665 TI - Manipulation of cracks in three-dimensional colloidal crystal films via recognition of surface energy patterns: an approach to regulating crack patterns and shaping microcrystals. AB - A new concept for dealing with cracks in colloidal crystals has been proposed. We induce the cracks rather than eliminate them via templates that possess hydrophilic/hydrophobic patterns on the surface (surface energy patterns), leading the cracks to propagate along the predetermined lines. Colloidal crystal arrays with various kinds of element figures separated by cracks could be reproducibly fabricated. Diverse crack patterns other than common stripes have been observed, and the mechanism of these behaviors has been explored. The factors that influence the crack density have been investigated to ensure that the templates could function effectively. Moreover, we obtained microcrystal blocks with specific shapes, detached from the substrate. PMID- 21630666 TI - Optimization of 1,2,3,4-tetrahydroacridin-9(10H)-ones as antimalarials utilizing structure-activity and structure-property relationships. AB - Antimalarial activity of 1,2,3,4-tetrahydroacridin-9(10H)-ones (THAs) has been known since the 1940s and has garnered more attention with the development of the acridinedione floxacrine (1) in the 1970s and analogues thereof such as WR 243251 (2a) in the 1990s. These compounds failed just prior to clinical development because of suboptimal activity, poor solubility, and rapid induction of parasite resistance. Moreover, detailed structure-activity relationship (SAR) studies of the THA core scaffold were lacking and SPR studies were nonexistent. To improve upon initial findings, several series of 1,2,3,4-tetrahydroacridin-9(10H)-ones were synthesized and tested in a systematic fashion, examining each compound for antimalarial activity, solubility, and permeability. Furthermore, a select set of compounds was chosen for microsomal stability testing to identify physicochemical liabilities of the THA scaffold. Several potent compounds (EC(50) < 100 nM) were identified to be active against the clinically relevant isolates W2 and TM90-C2B while possessing good physicochemical properties and little to no cross resistance. PMID- 21630667 TI - Initial efforts toward the optimization of arylomycins for antibiotic activity. AB - While most clinically used antibiotics were derived from natural products, the isolation of new broad-spectrum natural products has become increasingly rare and narrow-spectrum agents are typically deemed unsuitable for development because of intrinsic limitations of their scaffold or target. However, it is possible that the spectrum of a natural product antibiotic might be limited by specific resistance mechanisms in some bacteria, such as target mutations, and the spectra of such "latent" antibiotics might be reoptimized by derivatization, just as has been done with clinically deployed antibiotics. We recently showed that the spectrum of the arylomycin natural product antibiotics, which act via the novel mechanism of inhibiting type I signal peptidase, is broader than previously believed and that resistance in several key human pathogens is due to the presence of a specific Pro residue in the target peptidase that disrupts interactions with the lipopeptide tail of the antibiotic. To begin to test whether this natural resistance might be overcome by derivatization, we synthesized analogues with altered lipopeptide tails and identified several with an increased spectrum of activity against S. aureus. The data support the hypothesis that the arylomycins are latent antibiotics, suggest that their spectrum may be optimized by derivatization, and identify a promising scaffold upon which future optimization efforts might focus. PMID- 21630668 TI - UCST-like hybrid PAAm-AA/Fe3O4 microgels. Effect of Fe3O4 nanoparticles on morphology, thermosensitivity and elasticity. AB - The incorporation of metal oxide nanoparticles into microgels forming hybrid systems gives additional functionalities to the system and widens the field of potential application in biomedicine, biotechnology, and other fields. In particular, there have been very few investigations regarding UCST-like hybrid microgels. In connection with this, we report the preparation of UCST-like hybrid microgels of magnetite nanoparticles (Fe(3)O(4)) encapsulated in poly(acrylamide acrylic acid) microgel matrix via an inverse emulsion polymerization method. The key factor in the preparation of hybrid microgels is the need to divide in two the aqueous phase of the emulsion and feed them separately in order to avoid the aggregation of magnetic nanoparticles prior to polymerization reaction. The morphology, size, and spherical shape of hybrid microgels are determined by scanning electron microscopy. The encapsulation of magnetite nanoparticles within the polymer matrix is confirmed by transmission electron microscopy. Dynamic light scattering is employed to study both the swelling UCST-like behavior and the surface charge of the hybrid microgels. Swelling measurements confirm that the incorporation of magnetite does not affect the thermosensitivity of the system. In order to highlight the rheological behavior that can affect the final potential applications of these hybrid systems, a deep study of the viscoelastic properties is carried out by means of an oscillatory rheometer. The dependence of G' and G'' of the microgel dispersions with the frequency suggests a gel-like behavior and hence the occurrence of structural organization. In order to understand this structure formation and the influence of the magnetite in the interaction between hybrid microgels, scaling theory was applied. In terms of rheology, the addition of magnetite leads to a change in the interaction between hybrid microgels giving rise to an increase in the elasticity of the system. PMID- 21630669 TI - Effect of structural modification of alpha-aminoxy peptides on their intestinal absorption and transport mechanism. AB - A representative alpha-aminoxy peptide 1 has been demonstrated to have a potential for the treatment of human diseases associated with Cl(-) channel dysfunctions. However, its poor intestinal absorption was determined. The purpose of this study was to delineate the transport mechanism responsible for its poor absorption and also to prepare peptide analogues by structural modifications of 1 at its isobutyl side chains without changing the alpha-aminoxy core for retaining biological activity to improve the intestinal absorption. The poor intestinal absorption of 1 was proved to be due to the P-glycoprotein (P-gp) mediated efflux transport in Caco-2 cell monolayer, intestinal segments in Ussing chamber and rat single pass intestinal perfusion models. Four analogues with propionic acid (2), butanamine (3), methyl (4) and hydroxymethyl side chains (5) were synthesized and tested using the same models. Except for the permeability of 2, the absorbable permeability of the modified peptides in Caco-2 cell monolayer and their intestinal absorption in rats were significantly improved to 7-fold (3), 4-fold (4), 11-fold (5) and 36-fold (2), 42-fold (3), 55-fold (4), 102-fold (5), respectively, compared with 1 (P(app), 0.034 +/- 0.003 * 10(-6) cm/s; P(blood), 1.61 +/- 0.807 * 10(-6) cm/s). More interestingly, the structural modification remarkably altered transport mechanism of the peptides, leading to the conversion of the active transport via P-gp mediation (1, 2), to MRP mediation (3), MRP plus BCRP mediation (4) or a passive diffusion (5). Furthermore, P-gp mediated efflux transport of 1 and 2 was demonstrated to not alter the P-gp expression, while 1 but not 2 exhibited uncompetitive inhibitory effect on P-gp ATPase. The results demonstrated that intestinal absorption and transport mechanism of the alpha aminoxy peptides varied significantly with different structures, and their absorption can be dramatically improved by structural modifications, which allow us to further design and prepare better alpha-aminoxy peptide candidates with appropriate pharmacokinetic fates, including intestinal absorption, for potential clinical use. PMID- 21630671 TI - Ratiometric fluorescent receptors for both Zn2+ and H2PO4(-) ions based on a pyrenyl-linked triazole-modified homooxacalix[3]arene: a potential molecular traffic signal with an R-S latch logic circuit. AB - A ratiometric fluorescent receptor with a C(3) symmetric structure based on a pyrene-linked triazole-modified homooxacalix[3]arene (L) was synthesized and characterized. This system exhibited an interesting ratiometric detection signal output for targeting cations and anions through switching the excimer emission of pyrene from the "on-off" to the "off-on" type in neutral solution. (1)H NMR titration results suggested that the Zn(2+) center of receptor L.Zn(2+) provided an excellent pathway of organizing anion binding groups for optimal host-guest interactions. It is thus believed that this receptor has potential application in sensing, detection, and recognition of both Zn(2+) and H(2)PO(4)(-) ions with different optical signals. In addition, the fluorescence emission changes by the inputs of Zn(2+) and H(2)PO(4)(-) ions can be viewed as a combinational R-S latch logic circuit at the molecular level. PMID- 21630670 TI - A 3-in-1 polymeric micelle nanocontainer for poorly water-soluble drugs. AB - Poly(ethylene glycol)-block-poly(D,L-lactic acid) (PEG-b-PLA) micelles have a proven capacity for drug solubilization and have entered phase III clinical trials as a substitute for Cremophor EL in the delivery of paclitaxel in cancer therapy. PEG-b-PLA is less toxic than Cremophor EL, enabling a doubling of paclitaxel dose in clinical trials. We show that PEG-b-PLA micelles act as a 3-in 1 nanocontainer for paclitaxel, 17-allylamino-17-demethoxygeldanamycin (17-AAG), and rapamycin for multiple drug solubilization. 3-in-1 PEG-b-PLA micelles were ca. 40 nm in diameter; dissolved paclitaxel, 17-AAG, and rapamycin in water at 9.0 mg/mL; and were stable for 24 h at 25 degrees C. The half-life for in vitro drug release (t(1/2)) for 3-in-1 PEG-b-PLA micelles was 1-15 h under sink conditions and increased in the order of 17-AAG, paclitaxel, and rapamycin. The t(1/2) values correlated with log P(o/w) values, implicating a diffusion controlled mechanism for drug release. The IC(50) value of 3-in-1 PEG-b-PLA micelles for MCF-7 and 4T1 breast cancer cell lines was 114 +/- 10 and 25 +/- 1 nM, respectively; combination index (CI) analysis showed that 3-in-1 PEG-b-PLA micelles exert strong synergy in MCF-7 and 4T1 breast cancer cell lines. Notably, concurrent intravenous (iv) injection of paclitaxel, 17-AAG, and rapamycin using 3-in-1 PEG-b-PLA micelles was well-tolerated by FVB albino mice. Collectively, these results suggest that PEG-b-PLA micelles carrying paclitaxel, 17-AAG, and rapamycin will provide a simple yet safe and efficacious 3-in-1 nanomedicine for cancer therapy. PMID- 21630672 TI - Investigations of the catalytic mechanism of thioredoxin glutathione reductase from Schistosoma mansoni. AB - Thioredoxin glutathione reductase from Schistosoma mansoni (SmTGR) catalyzes the reduction of both thioredoxin and glutathione disulfides (GSSG), thus playing a crucial role in maintaining redox homeostasis in the parasite. In line with this role, previous studies have demonstrated that SmTGR is a promising drug target for schistosomiasis. To aid in the development of efficacious drugs that target SmTGR, it is essential to understand the catalytic mechanism of SmTGR. SmTGR is a dimeric flavoprotein in the glutathione reductase family and has a head-to-tail arrangement of its monomers; each subunit has the components of both a thioredoxin reductase (TrxR) domain and a glutaredoxin (Grx) domain. However, the active site of the TrxR domain is composed of residues from both subunits: FAD and a redox-active Cys-154/Cys-159 pair from one subunit and a redox-active Cys 596'/Sec-597' pair from the other; the active site of the Grx domain contains a redox-active Cys-28/Cys-31 pair. Via its Cys-28/Cys-31 dithiol and/or its Cys 596'/Sec-597' thiol-selenolate, SmTGR can catalyze the reduction of a variety of substrates by NADPH. It is presumed that SmTGR catalyzes deglutathionylation reactions via the Cys-28/Cys-31 dithiol. Our anaerobic titration data suggest that reducing equivalents from NADPH can indeed reach the Cys-28/Cys-31 disulfide in the Grx domain to facilitate reductions effected by this cysteine pair. To clarify the specific chemical roles of each redox-active residue with respect to its various reactivities, we generated variants of SmTGR. Cys-28 variants had no Grx deglutathionylation activity, whereas Cys-31 variants retained partial Grx deglutathionylation activity, indicating that the Cys-28 thiolate is the nucleophile initiating deglutathionylation. Lags in the steady-state kinetics, found when wild-type SmTGR was incubated at high concentrations of GSSG, were not present in Grx variants, indicating that this cysteine pair is in some way responsible for the lags. A Sec-597 variant was still able to reduce a variety of substrates, albeit slowly, showing that selenocysteine is important but is not the sole determinant for the broad substrate tolerance of the enzyme. Our data show that Cys-520 and Cys-574 are not likely to be involved in the catalytic mechanism. PMID- 21630673 TI - Duplex end breathing determines serum stability and intracellular potency of siRNA-Au NPs. AB - Structural requirements of siRNA-functionalized gold nanoparticles (siRNA-Au NPs) for Dicer recognition and serum stability were studied. We show that the 3' overhang on the nucleic acids of these particles is preferentially recognized by Dicer but also makes the siRNA duplexes more susceptible to nonspecific serum degradation. Dicer and serum nucleases show lower preference for blunt duplexes as opposed to those with 3' overhangs. Importantly, gold nanoparticles functionalized with blunt duplexes with relatively less thermal breathing are up to 15 times more stable against serum degradation without compromising Dicer recognition. This increased stability leads to a 300% increase in cellular uptake of siRNA-Au NPs and improved gene knockdown. PMID- 21630674 TI - Characterization of interactions between PilA from Pseudomonas aeruginosa strain K and a model membrane. AB - Type IV pili are important adhesion and motility factors in both gram-negative and gram-positive bacterial species, making pilus assembly from pilin subunits an important biophysical mechanism to understand at an atomic level. Knowledge of the pilus assembly mechanism has applications in antibiotic development, microbial physiology, and systems biology. We applied molecular dynamics simulations to investigate the position and orientation of Pseudomonas aeruginosa strain K PilA in a model membrane as well as the interactions of PilA with the surrounding lipids, identifying several key residues that stabilize the position of PilA within the membrane. Furthermore, we performed umbrella sampling to determine the free energy of extracting this protein from the membrane. Our results provide insight into the molecular events and energetics associated with pilus assembly, thereby initiating a detailed examination of the dynamics of this process. PMID- 21630675 TI - Observations of host guest interactions specific to molecular matrices: water monomers and dimers in hydrogen matrices. AB - Water monomers and dimers have been studied at low temperatures in matrices of solid p-H(2), o-D(2), n-H(2), and n-D(2) using infrared spectroscopy. Our data demonstrate interaction mechanisms between host matrix and guest molecules that are different from the ones observed in atomic noble gas matrices. Notably both guest/host rotational--rotational interaction and matrix induced modifications of the guest libration modes are observed. We also show that different types of interaction influence the relaxation times of some of guest modes. Water rotates freely in p-H(2) and o-D(2) but librates in n-H(2) and n-D(2). Rotational relaxation is faster in o-D(2) than in p-H(2) and faster in p-H(2) than in Ne. This is attributed to interactions between water rotation and matrix molecule rotation in p-H(2) and o-D(2). In n-H(2) and n-D(2), a strong water libration band is observed in the far-infrared, and strong water monomer vibration bands have libration satellites. Water dimer bands, close to matrix rotation bands, are perturbed by the matrix motions. The H-bonded isomer H(2)O--HOD rapidly converts to the D-bonded form H(2)O-DOH in p-H(2) and in o-D(2) but slowly in n-H(2) and n D(2). PMID- 21630676 TI - Character and chlorine reactivity of dissolved organic matter from a mountain pine beetle impacted watershed. AB - Lodgepole pine needle leachates from trees killed by the mountain pine beetle epidemic in Colorado were evaluated for dissolved organic matter (DOM) character, biodegradation, treatability by coagulation and disinfection byproduct (DBP) formation. An average of 8.0 (+/-0.62) mg-DOC/g-dry weight of litter was leached from three sets of needle samples representing different levels of forest floor degradation. Fluorescence analysis included collection of excitation and emission matrices, examination of peak intensities and development of a 4-component parallel factor (PARAFAC) analysis model. Peak intensity and PARAFAC analyses provided complementary results showing that fresh leachates were initially dominated by polyphenolic/protein-like components (60-70%) and humic-like fluorescence increased (40-70%) after biodegradation. Humic-like components were removed by coagulation (20-64%), while polyphenolic/protein-like components were not, which may create challenges for utilities required to meet OM removal regulations. DBP formation yields after 24 h chlorination were 20.5-26.4 MUg/mg DOC for trihalomethanes and 9.0-14.5 MUg/mg-DOC for haloacetic acids for fresh leachates; increased after biodegradation to 19.2-64.2 and 7.1-30.9 MUg/mg-DOC, respectively; and decreased after coagulation (fresh: 11.3-17.7;5.7-7.6 MUg/mg DOC, respectively; biodegraded: 12.0-27.3 and 2.9-7.2 MUg/mg-DOC, respectively), reflective of changes in concentration of humic material. Humic-like PARAFAC components and peak intensities were positively correlated (R(2) >= 0.45) to DBP concentrations, while polyphenolic/protein-like components were not (R(2) <= 0.17). PMID- 21630677 TI - The electronic spectra of the sandwich stacked PFBT: a theoretical study. AB - Stacked models that include 9,9'-bis(6''-N,N,N-trimethylammonium)hexyl]fluorene co-alt-4,7-(2,1,3-benzothiadiazole)dibromide (F(BT)F) monomer sandwiched between two stacked 2,1,3-benzothiadiazole (BT) units were explored using theoretical approaches. Molecular structures and the optical characteristics of the investigated species were investigated at the M06-2X/6-311G(d,p)//TD-M06-2X/6 311G(d,p) level of theory. In all models, the electronic excitation to the lowest singlet pipi* excited state (S1(pipi*)) is governed by the highest occupied molecular orbital to lowest unoccupied molecular orbital (HOMO -> LUMO) transitions. The obtained results suggest that stacking interaction might have only minor effects on the transition energy for both absorption and emission processes. Instead, the reduction in the excitation energy of the stacked complexes should be attributed to the dipole-dipole interaction. The larger the interaction energy of the stacked models, the bigger the observed differences between absorption-emission energies. The presence of the solvation medium with small dielectric constant may increase the absorption-emission energy differences. It is expected that the largest absorption-emission shift can be observed in the benzene solution. PMID- 21630678 TI - Polyvalent nucleic acid nanostructures. AB - Polyvalent oligonucleotide-nanoparticle conjugates possess several unique emergent properties, including enhanced cellular uptake, high antisense bioactivity, and nuclease resistance, which hypothetically originate from the dense packing and orientation of oligonucleotides on the surface of the nanoparticle. In this Communication, we describe a new class of polyvalent nucleic acid nanostructures (PNANs), which are comprised of only cross-linked and oriented nucleic acids. We demonstrate that these particles are capable of effecting high cellular uptake and gene regulation without the need of a cationic polymer co-carrier. The PNANs also exhibit cooperative binding behavior and nuclease resistance properties. PMID- 21630679 TI - Enhanced thermopower via carrier energy filtering in solution-processable Pt Sb2Te3 nanocomposites. AB - This work demonstrates the first solution-processable metal-semiconductor nanocomposites with enhanced thermoelectric properties via carrier energy filtering. Platinum nanocrystals are embedded in a p-type antimony(III) telluride (Sb(2)Te(3)) semiconductor matrix, thus introducing band-bending potentials for holes. By scattering low energy holes, an increase in thermopower is observed. Introduction of Pt nanocrystals also increases carrier concentration thereby partially compensating for reduced electrical conductivity due to the decreased mobility. At room temperature, an improvement in thermoelectric power factor was achieved compared to that of the Sb(2)Te(3) films. This work highlights the possibility of combining a diverse set of n- and p-type semiconductor matrices with nanocrystals to engineer and optimize energy-dependent carrier scattering with the ease of materials processing. PMID- 21630680 TI - Synthesis, crystal structures, and supramolecular assemblies of pyrrole-based anion receptors bearing modified pyrrole beta-substituents. AB - Dipyrrolyldiketone BF(2) complexes acting as acyclic anion receptors form supramolecular assemblies with structures and properties that are dependent on the pyrrole beta-substituents. In particular, although beta-alkyl substituents interfered with the formation of stable gel states, the introduction of fluorine moieties induced a stable supramolecular gel when compared to that of beta unsubstituted receptor. PMID- 21630681 TI - Synthesis and characterization of a BODIPY conjugate of the BCR-ABL kinase inhibitor Tasigna (nilotinib): evidence for transport of Tasigna and its fluorescent derivative by ABC drug transporters. AB - Tasigna (Nilotinib) is a BCR-ABL kinase inhibitor recently approved by the Food and Drug Administration, which is indicated for the treatment of drug-resistant chronic myelogenous leukemia (CML). The efflux of tyrosine kinase inhibitors by ATP-binding cassette (ABC) drug transporters, which actively pump these drugs out of cells utilizing ATP as an energy source, has been linked to the development of drug resistance in CML patients. We report here the synthesis and characterization of a fluorescent derivative of Tasigna to study its interaction with two major ABC transporters, P-glycoprotein (Pgp) and ABCG2, in in vitro and ex vivo assays. A fluorescent derivative of Tasigna, BODIPY FL Tasigna, inhibited the BCR-ABL kinase activity in K562 cells and was also effluxed by Pgp- and ABCG2 expressing cells in both cultured cells and rat brain capillaries expressing Pgp and ABCG2. In addition, [(3)H]-Tasigna was found to be transported by Pgp expressing polarized LLC-PK1 cells in a transepithelial transport assay. Consistent with these results, both Tasigna and BODIPY FL Tasigna were less effective at inhibiting the phosphorylation of Crkl (a substrate of BCR-ABL kinase) in Pgp- and ABCG2-expressing K562 cells due to their reduced intracellular concentration. Taken together, these data provide evidence that BODIPY FL Tasigna is transported by Pgp and ABCG2, and Tasigna is transported by Pgp. Further, we propose that BODIPY FL Tasigna can potentially be used as a probe for functional analysis of Pgp and ABCG2 in cancer cells and in other preclinical studies. PMID- 21630682 TI - Impact of silica nanoparticle design on cellular toxicity and hemolytic activity. AB - Understanding the toxicity of silica nanoparticles (SiO(2)) on the cellular level is crucial for rational design of these nanomaterials for biomedical applications. Herein, we explore the impacts of geometry, porosity, and surface charge of SiO(2) on cellular toxicity and hemolytic activity. Nonporous Stober silica nanospheres (115 nm diameter), mesoporous silica nanospheres (120 nm diameter, aspect ratio 1), mesoporous silica nanorods with aspect ratio of 2, 4, and 8 (width by length 80 * 200 nm, 150 * 600 nm, 130 * 1000 nm), and their cationic counterparts were evaluated on macrophages, lung carcinoma cells, and human erythrocytes. It was shown that the toxicity of SiO(2) is cell-type dependent and that surface charge and pore size govern cellular toxicity. Using inductively coupled plasma mass spectrometry, the cellular association of SiO(2) was quantitated with the association amount increasing in the following order: mesoporous SiO(2) (aspect ratio 1, 2, 4, 8) < amine-modified mesoporous SiO(2) (aspect ratio 1, 2, 4, 8) < amine-modified nonporous Stober SiO(2) < nonporous Stober SiO(2). Geometry did not seem to influence the extent of SiO(2) association at early or extended time points. The level of cellular association of the nanoparticles was directly linked to the extent of plasma membrane damage, suggesting a biological cause-and-effect relationship. Hemolysis assay showed that the hemolytic activity was porosity- and geometry-dependent for bare SiO(2) and surface-charge-dependent for amine-modified SiO(2). A good correlation between hemolytic activity and cellular association was found on a similar dosage basis. These results can provide useful guidelines for the rational design of SiO(2) in nanomedicine. PMID- 21630683 TI - Hybrid organo-inorganic clay with nonionic interlayers. mid- and near-IR spectroscopic studies. AB - The intercalation of organic polymers molecules (i.e., PEGs and BRIJ) into a standard Ca-montmorillonite has been studied by XRD, TG, and IR spectroscopy. The polymer intercalation is confirmed by the increasing of the d(001) in XRD spectra as well as by the complex multisteps thermal decomposition behavior of the organo clay materials. Mid-IR and diffuse reflectance near-IR spectra of the intercalated materials show the polymer diagnostic bands (CH stretching and deformation mode), shifted or changed in shape by the interaction with the clay matrix. Both PEG 1500 and PEG 4000 based materials are likely intercalated in an extended configuration, similar to the amorphous polymer form. BRIJ intercalated polymer spectra suggest the disordered conformation of the alkilic chain in a prevailing "gauche", poorly packed, conformation. Host montmorillonite IR bands, mainly OH and water stretching and deformation fundamentals, combination, and overtone bands, are reduced in intensity by polymer intercalation, pointing out an interaction, likely through H-bonding and/or a possible substitution of cations hydration water molecules. PMID- 21630684 TI - Langendorff heart: a model system to study cardiovascular effects of engineered nanoparticles. AB - Engineered nanoparticles (ENPs) are produced and used in increasing quantities for industrial products, food, and drugs. The fate of ENPs after usage and impact on health is less known. Especially as air pollution, suspended nanoparticles have raised some attention, causing diseases of the lung and cardiovascular system. Human health risks may arise from inhalation of ENPs with associated inflammation, dispersion in the body, and exposure of vulnerable organs (e.g., heart, brain) and tissues with associated toxicity. However, underlying mechanisms are largely unknown. Furthermore future use of ENPs in therapeutic applications is being researched. Therefore knowledge about potential cardiovascular risks due to exposure to ENPs is highly demanded, but there are no established biological testing models yet. Therefore, we established the isolated beating heart (Langendorff heart) as a model system to study cardiovascular effects of ENPs. This model enables observation and analysis of electrophysiological parameters over a minimal time period of 4 h without influence by systemic effects and allows the determination of stimulated release of substances under influence of ENPs. We found a significant dose and material dependent increase in heart rate accompanied by arrhythmia evoked by ENPs made of flame soot (Printex 90), spark discharge generated soot, anatas (TiO(2)), and silicon dioxide (SiO(2)). However, flame derived SiO(2) (Aerosil) and monodisperse polystyrene lattices exhibited no effects. The increase in heart rate is assigned to catecholamine release from adrenergic nerve endings within the heart. We propose the isolated Langendorff heart and its electrophysiological characterization as a suitable test model for studying cardiovascular ENP toxicity. PMID- 21630685 TI - Synthesis of molybdenum nitrido complexes for triple-bond metathesis of alkynes and nitriles. AB - Complexes of the type N=Mo(OR)(3) (R = tertiary alkyl, tertiary silyl, bulky aryl) have been synthesized in the search for molybdenum-based nitrile-alkyne cross-metathesis (NACM) catalysts. Protonolysis of known N=Mo(NMe(2))(3) led to the formation of N=Mo(O-2,6-(i)Pr(2)C(6)H(3))(3)(NHMe(2)) (12), N=Mo(OSiPh(3))(3)(NHMe(2)) (5-NHMe(2)), and N=Mo(OCPh(2)Me)(3)(NHMe(2)) (17 NHMe(2)). The X-ray structure of 12 revealed an NHMe(2) ligand bound cis to the nitrido ligand, while 5-NHMe(2) possessed an NHMe(2) bound trans to the nitride ligand. Consequently, 17-NHMe(2) readily lost its amine ligand to form N=Mo(OCPh(2)Me)(3) (17), while 12 and 5-NHMe(2) retained their amine ligands in solution. Starting from bulkier tris-anilide complexes, N=Mo(N[R]Ar)(3) (R = isopropyl, tert-butyl; Ar = 3,5-dimethylphenyl) allowed for the formation of base free complexes N=Mo(OSiPh(3))(3) (5) and N=Mo(OSiPh(2)(t)Bu)(3) (16). Achievement of a NACM cycle requires the nitride complex to react with alkynes to form alkylidyne complexes; therefore the alkyne cross-metathesis (ACM) activity of the complexes was tested. Complex 5 was found to be an efficient catalyst for the ACM of 1-phenyl-1-butyne at room temperature. Complexes 12 and 5-NHMe(2) were also active for ACM at 75 degrees C, while 17-NHMe(2) and 16 did not show ACM activity. Only 5 proved to be active for the NACM of anisonitrile, which is a reactive substrate in NACM catalyzed by tungsten. NACM with 5 required a reaction temperature of 180 degrees C in order to initiate the requisite alkylidyne-to nitride conversion, with slightly more than two turnovers achieved prior to catalyst deactivation. Known molybdenum nitrido complexes were screened for NACM activity under similar conditions, and only N=Mo(OSiPh(3))(3)(py) (5-py) displayed any trace of NACM activity. PMID- 21630686 TI - Aggregation kinetics of citrate and polyvinylpyrrolidone coated silver nanoparticles in monovalent and divalent electrolyte solutions. AB - The aggregation kinetics of silver nanoparticles (AgNPs) that were coated with two commonly used capping agents-citrate and polyvinylpyrrolidone (PVP)--were investigated. Time-resolved dynamic light scattering (DLS) was employed to measure the aggregation kinetics of the AgNPs over a range of monovalent and divalent electrolyte concentrations. The aggregation behavior of citrate-coated AgNPs in NaCl was in excellent agreement with the predictions based on Derjaguin Landau-Verwey-Overbeek (DLVO) theory, and the Hamaker constant of citrate-coated AgNPs in aqueous solutions was derived to be 3.7 * 10(-20) J. Divalent electrolytes were more efficient in destabilizing the citrate-coated AgNPs, as indicated by the considerably lower critical coagulation concentrations (2.1 mM CaCl(2) and 2.7 mM MgCl(2) vs 47.6 mM NaCl). The PVP-coated AgNPs were significantly more stable than citrate-coated AgNPs in both NaCl and CaCl(2), which is likely due to steric repulsion imparted by the large, noncharged polymers. The addition of humic acid resulted in the adsorption of the macromolecules on both citrate- and PVP-coated AgNPs. The adsorption of humic acid induced additional electrosteric repulsion that elevated the stability of both nanoparticles in suspensions containing NaCl or low concentrations of CaCl(2). Conversely, enhanced aggregation occurred for both nanoparticles at high CaCl(2) concentrations due to interparticle bridging by humic acid aggregates. PMID- 21630687 TI - Biomimetic cationic polyannulation reaction catalyzed by Bi(OTf)3: cyclization of 1,6-dienes, 1,6,10-trienes, and aryl polyenes. AB - Nonactivated trienes and aryltrienes were cyclized into polycyclic compounds in good to excellent yields under bismuth triflate catalysis in a biomimetic fashion. The reaction showed broad applicability and allowed for the formation of functionalized bicyclic to tetracyclic structures from simple precursors in one pot. For some specific substrates, the cyclization was followed by a methyl shift as encountered in terpenoid biosynthesis. PMID- 21630688 TI - Is indirect exposure a significant contributor to the burden of perfluorinated acids observed in humans? AB - In comparison to other persistent organic pollutants, human fluorochemical contamination is relatively complicated. This complication arises at least in part from a disparity between the chemicals used commercially and those measured in the environment and humans. Commercial fluorochemical products are dominated by fluorinated polymers used in textile or carpet applications, or fluorosurfactants used in applications ranging from personal care products, leveling and wetting agents, to greaseproofing food-contact materials. Investigations into environmental and human fluorochemical contamination have focused on perfluorinated acids (PFAs), either the perfluorinated carboxylates (PFCAs) or sulfonates (PFSAs). In this review we will present an overview of data related to human fluorochemical exposure including a discussion of fluorochemical production, concentrations in exposure media, biotransformation processes producing PFAs, and trends in human sera. These data will be presented in the context of how they can inform sources of human PFA contamination, specifically whether the contamination results from direct PFA exposure or indirect exposure via the biotransformation of commercial fluorochemicals or their residuals. Concentrations of both perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA) began to decrease in human sera around the year 2000, a change that mirrored the 2000-2002 phase-out of perfluorooctane sulfonyl fluoride (POSF) production. These temporal trends suggest exposure to current-use POSF-based materials was a significant source of PFOA and PFOS exposure prior to 2000. Relatively slow PFOA elimination and increasing concentrations of the C9 and C10 PFCAs in human sera suggest continued PFCA exposure, without similar exposure to PFOS, which is consistent with indirect exposure via the biotransformation of fluorotelomer-based materials. Conversely, human exposure models have suggested direct exposure to PFAs present in food items is the major source of human contamination. The data set presented here cannot unequivocally delineate between direct and indirect human exposure, however temporal trends in human sera and exposure media are consistent with indirect exposure representing a significant portion of observed human PFA contamination. PMID- 21630689 TI - Ruthenium-catalyzed hydrovinylation of N-acetylenamines leading to amines with a quaternary carbon center. AB - A catalytic hydrovinylation of N-acetylenamines with ethylene is reported. This new hydrovinylation reaction is catalyzed by a ruthenium hydride complex, RuHCl(CO)(PCy(3))(2), providing a series of N-acetylamines with a quaternary carbon center with up to 99% yield. PMID- 21630690 TI - Novel organometallic reagents: geminal dianionic derivatives of the heavy group 14 elements. AB - This Forum review describes the most recent achievements in the novel prospective field of highly reactive main-group organometallics, namely, geminal dianionic derivatives of the heavy group 14 elements (Si, Ge, Sn). A brief historical introduction to the topic is followed by discussion of the current state of affairs in the field of stable derivatives and prospects for future efforts, highlighting our own synthetic approach and recent results. The most important experimental contributions, including synthesis of 1,1-dilithiosilane, -germane, and -stannane derivatives; dilithio(halo)silanes (lithiosilylenoids); metallole 1,1-dianions; and heavy analogues of the cyclobutadiene dianion derivatives, are presented, along with a discussion of the synthetic applications of the above mentioned organometallic compounds. PMID- 21630691 TI - Hydrophobic transition in porous amorphous silica. AB - Realistic models of amorphous silica surfaces with different silanol densities are built using Monte Carlo annealing. Water-silica interfaces are characterized by their energy interaction maps, adsorption isotherms, self-diffusion coefficients, and Poiseuille flows. A hydrophilic to hydrophobic transition appears as the surface becomes purely siliceous. These results imply significant consequences for the description of surfaces. First, realistic models are required for amorphous silica interfaces. Second, experimental amorphous silica hydrophilicity is attributed to charged or uncharged defects, and not to amorphousness. In addition, autoirradiation in nuclear waste glass releases hydrogen atoms from silanol groups and can induce such a transition. PMID- 21630692 TI - Analysis of the reactivity on the c7h6 potential energy surface. AB - The reactivity and decomposition kinetics on the C(7)H(6) potential energy surface (PES) were investigated, determining structures of stationary points at the B3LYP/6-31+G(d,p) level and energies at the CCSD(T)/cc-pVTZ level with extension to the complete basis set limit. For the reactions characterized by a significant multireference character, the energies were calculated at the CASPT2/cc-pVTZ level. The portion of the PES investigated consisted of 27 wells connected by 39 saddle points. Of the 27 wells, 16 can be accessed through transition states having activation energies smaller than the dissociation threshold. In agreement with previous theoretical studies, it was found that the main interconversion channel takes place on the singlet PES and connects phenylcarbene, cycloheptatetrane, spiroheptatriene, fulvenallene, and three ethynylcyclopentadiene isomers. Two new mechanisms are proposed for the formation of 5-ethynylcyclopentadiene and for the conversion of spiroheptatriene to fulvenallene. The unimolecular decomposition kinetics was thoroughly investigated. It was found that the fastest high pressure decomposition channel, at the temperatures at which C(7)H(6) undergoes unimolecular decomposition (1500- 2000 K), leads to the formation of cyclopentadienylidene and acetylene. The rate of crossing from the singlet to the triplet PES may affect considerably this reaction channel, as it is formally spin forbidden. The alternative pathway, which is the decomposition to fulvenallenyl, is however only a factor of 2--3 slower and significantly less activated (82 vs 96 kcal/mol). PMID- 21630693 TI - Origin of buckling phenomenon during drying of micrometer-sized colloidal droplets. AB - The origin of the buckling of micrometer-sized colloidal droplets during evaporation-induced self-assembly (EISA) has been elucidated using electron microscopy and small-angle neutron scattering. Doughnut-like assembled grains with varying aspect ratios are formed during EISA at different physicochemical conditions. It has been revealed that this phenomenon is better explained by an existing hypothesis based on the formation of a viscoelastic shell of nanoparticles during drying than by other existing hypotheses based on the inertial instability of the initial droplets and hydrodynamic instability due to thermocapillary forces. This conclusion was further supported by the arrest of buckling through modification of the colloidal interaction in the initial dispersion. PMID- 21630694 TI - Measurement of free Cu ion activity in seawater using a passive-equilibrium sonic assisted free ion recorder (SAFIR). AB - A passive equilibrium probe, sonic-assisted free ion recorder (SAFIR), was recently developed to monitor free Cu ion activity ({Cu(2+)}) in aquatic environments. The SAFIR is user-friendly and can be deployed on-site to record the ambient {Cu(2+)} in an accurate and fast manner. It consists of a chemically modified polystyrene disk (Primaria disk) containing metal ion-binding functional groups, coupled with a sonic horn in a flow-through cell. The calibration of the disk was successfully conducted in metal-ion buffered seawater solutions (pH 8.2, salinity = 32) with {Cu(2+)} ranging from 10(-9.0) to 10(-15.0) mol/L. The Cu adsorbed on the surface of the Primaria disk (Cu(ads)) correlated linearly with the ambient {Cu(2+)} through a Frendlich equilibrium adsorption model. The adsorption process was kinetically enhanced by the introduction of ultrasound, allowing equilibrium between the SAFIR and the ambient {Cu(2+)} in 10 min. The SAFIR was deployed in several coastal regions, and the {Cu(2+)} recorded by SAFIR were in good agreement with the results obtained from corresponding electrochemical analysis. PMID- 21630696 TI - Membrane morphology modifications induced by hydroquinones. AB - We synthesize and characterize alkylthiohydroquinones (ATHs) in order to investigate their interactions with lipid model membranes, POPE and POPC. We observe the formation of structures with different morphologies, or curvature of the lipid bilayer, depending on pH and increasing temperature. We attribute their formation to changes in the balance charge/polarity induced by the ATHs. Mixtures of ATHs with POPE at pH 4 form two cubic phases, P4(3)32 and Im3m, that reach a maximum lattice size at 40 degrees C while under basic conditions these phases only expand upon heating from room temperature. The cubic phases coexist with lamellar or hexagonal phases and are associated with inhomogeneous distribution of the ATH molecules over the lipid matrix. The zwitterionic POPC does not form cubic phases but instead shows lamellar structures with no clear influence of the 2,6-BATH. PMID- 21630697 TI - Potential energy surfaces for oxygen adsorption, dissociation, and diffusion at the Pt(321) surface. AB - We report a first-principles, periodic supercell analysis of oxygen adsorption, diffusion, and dissociation at the kinked Pt(321) surface. Binding energies and binding site preferences of isolated oxygen atoms and molecules have been determined, and we show that both atomic and molecular oxygen prefer binding in bridge sites involving coordinatively unsaturated kink Pt atoms. Binding energies of atomic and molecular oxygen in different sites correlate well with the average metallic Pt coordination number of Pt atoms forming each site, although differences exist between adsorbates in symmetrically similar sites due to the inherent chirality of the surface. Atomic O in the strongest binding bridge sites experiences relatively small energy barriers for diffusion to neighboring sites compared to O on Pt(111). However, due to the structure of the surface, O diffusion is only rapid between different sites around the kink Pt atom, whereas the effective long-range tracer diffusion, as determined from a simple course grain model, is shown to be anisotropic and slower than on the Pt(111) surface. Four dissociation pathways for O(2) at low coverage are also reported and found to be in agreement with experimental observations of facile dissociation, even at low temperature. PMID- 21630698 TI - Synthesis of 3-substituted indazoles from arynes and N-tosylhydrazones. AB - Readily available, stable, and inexpensive N-tosylhydrazones react with arynes under mild reaction conditions to afford 3-substituted indazoles in moderate to good yields. The reaction appears to involve a dipolar cycloaddition of in situ generated diazo compounds and arynes. PMID- 21630699 TI - Dynamical and rheological properties of fluorinated surfactant films adsorbed at the pressurized CO2-H2O interface. AB - The dynamics of adsorption, interfacial tension, and rheological properties of two phosphocholine-derived partially fluorinated surfactants FnHmPC, designed to compensate for the weak CO(2)-surfactant tail interactions, were determined at the pressurized CO(2)-H(2)O interface. The two surfactants differ only by the length of the hydrocarbon spacer (5 CH(2) in F8H5PC and 11 CH(2) in F8H11PC) located between the terminal perfluoroalkyl chain and the polar head. The length of this spacer was found to have a critical impact on the adsorption kinetics and elasticity of the interfacial surfactant film. F8H5PC is soluble in both water and CO(2) phases and presents several distinct successive interfacial behaviors when bulk water concentration (C(W)) increases and displays a nonclassical isotherm shape. The isotherms of F8H5PC are similar for the three CO(2) pressures investigated and comprise four regimes. In the first regime, at low C(W), the interfacial tension is controlled by the organization that occurs between H(2)O and CO(2). The second regime corresponds to the adsorption of the surfactant as a monolayer until the CO(2) phase is saturated with F8H5PC, resulting in a first inflection point. In this regime, F8H5PC molecules reach maximal compaction and display the highest apparent interfacial elasticity. In the third regime, a second inflection is observed that corresponds to the critical micelle concentration of the surfactant in water. At the highest concentrations (fourth regime), the interfacial films are purely viscous and highly flexible, suggesting the capacity for this surfactant to produce water-in-CO(2) microemulsion. In this regime, surfactant adsorption is very fast and equilibrium is reached in less than 100 s. The behavior of F8H11PC is drastically different: it forms micelles only in the water phase, resulting in a classical Gibbs interface. This surfactant decreases the interfacial tension down to 1 mN/m and forms a strongly elastic interface. As this surfactant forms a very cohesive interface, it should be suitable for formulating stable water-in-CO(2) emulsions. The finding that the length of the hydrocarbon spacer in partially fluorinated surfactants can drastically influence film properties at the CO(2)-H(2)O interface should help control the formation of microemulsions versus emulsions and help elaborate a rationale for the design of surfactants specifically adapted to pressurized CO(2). PMID- 21630700 TI - Selective binding of carbamate pesticides by self-assembled monolayers of calix[4]arene lipoic acid: wettability and impedance dual-signal response. AB - A calix[4]arene lipoic acid (C4LA) was synthesized by click chemistry in 62% yield. It was immobilized on Au surfaces via self-assembly to offer C4LA Self Assembled Monolayers (SAMs). The SAMs show wettability and electrochemical impedance dual-signal response for methomyl with highly sensitivity and selectivity. PMID- 21630701 TI - Speciation, luminescence, and alkaline fluorescence quenching of 4-(2 methylbutyl)aminodipicolinic acid (H2MEBADPA). AB - 4-(2-Methylbutyl)aminodipicolinic acid (H(2)MEBADPA) has been synthesized and fully characterized in terms of aqueous phase protonation constants (pK(a)'s) and photophysical measurements. The pK(a)'s were determined by spectrophotometric titrations, utilizing a fully sealed titration system. Photophysical measurements consisted of room temperature fluorescence and frozen solution phosphorescence as well as quantum yield determinations at various pH, which showed that only fully deprotonated MEBADPA(2-) is appreciably emissive. The fluorescence of MEBADPA(2-) has been determined to be quenched by hydroxide and methoxide anions, most likely through base-catalyzed excited-state tautomerism or proton transfer. This quenching phenomenon has been quantitatively explored through steady-state and time-resolved fluorescence measurements. Utilizing the determined pK(a)s and quenching constants, the fluorescent intensity of MEBADPA(2-) has been successfully modeled as a function of pH. PMID- 21630702 TI - Heterogeneous reaction rates in an ionic liquid: quantitative results from two dimensional multiple population-period transient spectroscopy. AB - The hypotheses that ionic liquids are structurally heterogeneous at the molecular level and, even further, that this heterogeneity can transfer to the rates of reactions run in ionic liquids is being actively debated. Here, this hypothesis is tested using multiple population-period transient spectroscopy (MUPPETS), an emerging type of multidimensional measurement that resolves the kinetics of subensembles within a heterogeneous sample. A previous MUPPETS study of the excited-state twisting and electronic relaxation of auramine indicated that an ionic-liquid solvent induces rate dispersion due to a combination of heterogeneous and homogeneous processes, but those data could not quantitatively separate these contributions [Khurmi, C.; Berg, M. A. J. Phys. Chem. Lett.2010, 1, 161]. New MUPPETS data that include phase resolution and subtraction of thermal gratings are presented here and are successfully modeled. The total range of reaction rates (10--90%) is a factor of 70. If the solvent effect is viewed as a set of local viscosities, the viscosity distribution is broad and highly asymmetric. However, if the solvent is viewed as changing a reaction barrier, the data correspond to a Gaussian distribution of barrier heights. The relaxation of each subensemble is nonexponential with an initial induction period, but the shape of the decay is invariant across the rate distribution. A small (2%), long lived component is identified as a part of the homogeneous kinetic scheme and thus as a secondary channel for excited-state relaxation, not as an impurity or alternative ground-state form of auramine. On the basis of these results, we suggest that the primary cause of rate heterogeneity is a long-lived local electric field acting on the charge redistribution during the reaction. PMID- 21630703 TI - The singular gas-phase structure of 1-aminocyclopropanecarboxylic acid (Ac3c). AB - The natural nonproteinogenic alpha-amino acid 1-aminocyclopropanecarboxylic acid (Ac(3)c) has been vaporized by laser ablation and studied in the gas phase by molecular-beam Fourier transform microwave spectroscopy. Comparison of the experimental rotational and (14)N nuclear quadrupole coupling constants with the values predicted ab initio for these parameters has allowed the unambiguous identification of three Ac(3)c conformers differing in the hydrogen bonding pattern. Two of them resemble those characterized before for the coded aliphatic alpha-amino acids. Remarkably, a third conformer predicted to be energetically accessible for all of these amino acids but never observed (the so-called "missing conformer") has been found for Ac(3)c, close in energy to the global minimum. This is the first time that such a conformer, stabilized by an N H...O(H) hydrogen bond, is detected in the rotational spectrum of a gaseous alpha amino acid with a nonpolar side chain. The conjugative interaction established between the cyclopropane ring and the adjacent carbonyl group seems to be responsible for the unique conformational properties exhibited by Ac(3)c. PMID- 21630704 TI - Chemoenzymatic synthesis of inositols, conduritols, and cyclitol analogues. PMID- 21630705 TI - Novel tetrapeptide, RGDF, mediated tumor specific liposomal doxorubicin (DOX) preparations. AB - Arginine-glycine-aspartate (RGD) has been shown to possess a strong affinity for the integrins overexpressed in tumor cells, especially during tumor invasion, angiogenesis and metasis. Based on work from others, a novel tetrapeptide, arginine-glycine-aspartate-phenylanaline (RGDF), has been designed and studied as a homing device to direct liposomal doxorubicin (DOX) to tumor cells in this work. In order to incorporate RGDF into liposomal DOX preparations, RGDF was conjugated with three different fatty alcohols to achieve RGDF-fatty alcohol conjugates. Glycine-glycine-aspartate-phenylanaline (GGDF)-lauryl alcohol conjugate was synthesized as a negative control. RGDF-fatty alcohol conjugates (RGDFO(CH(2))(n)CH(3)) and GGDF-lauryl alcohol conjugate (L-GGDFC12-DOX) incorporated liposomal preparations were obtained by first preparing liposomes using the film dispersion method followed by loading DOX using a transmembrane pH gradient method. Because of their amphipathic nature, RGDF- or GGDF-fatty alcohol conjugates are expected to be readily incorporated into liposomes with their fatty alkanyl chains being intercalated between fatty acyl chains of liposomal bilayers and the hydrophilic peptide moiety (RGDF or GGDF) being anchored on the surface of liposomes. The particle size and zeta potential of liposomal DOX preparations containing RGDF-fatty alcohol conjugate (L-RGDF-DOXs) or L-GGDFC12 DOX were measured, and their morphology was studied using transmission electron microscopy. In vitro DOX release profile from RGDF incorporated liposomal DOX was measured. The antitumor activities of RGDF incorporated liposomal DOX preparations were evaluated in ICR mice inoculated with sarcoma S(180), which is known to express alpha(v)beta(3) integrin. Both conventional liposomal DOX preparation (L-DOX) without RGDFO(CH(2))(n)CH(3) and L-GGDFC12-DOX were used as negative controls. Our results showed improved tumor growth inhibition with L RGDF-DOXs over doxorubicin hydrochloride solution, L-DOX and L-GGDFC12-DOX. Pathological examination of tumor biopsy demonstrated that L-RGDF-DOXs induced enhanced tumor cell death in comparison to negative controls. Pharmacokinetic studies showed that the concentrations of DOX found in tumor sites were increased by 1.7-4.5-fold when liposomal DOX preparation containing RGDF-lauryl alcohol conjugate (L-RGDFC12-DOX) was administered in comparison to when L-GGDFC12-DOX or doxorubicin hydrochloride solution was administered. The concentrations of DOX found in the heart, which is the main site of toxic effects of DOX, were significantly reduced when L-RGDFC12-DOX was administered in comparison to when L GGDFC12-DOX or doxorubicin hydrochloride solution was administered. PMID- 21630707 TI - Donor-acceptor conjugated polymer based on naphtho[1,2-c:5,6 c]bis[1,2,5]thiadiazole for high-performance polymer solar cells. AB - Donor-acceptor conjugated polymers PBDT-DTBT and PBDT-DTNT, based on 2,1,3 benzothiadiazole (BT) and naphtho[1,2-c:5,6-c]bis[1,2,5]thiadiazole (NT), have been designed and synthesized for polymer solar cells. NT contains two fused 1,2,5-thiadiazole rings that lower the band gap, enhance the interchain packing, and improve the charge mobility of the resulting polymer. Consequently, the NT based polymer PBDT-DTNT exhibited considerably better photovoltaic performance with a power conversion efficiency (PCE) of 6.00% when compared with the BT-based polymer PBDT-DTBT, which gave a PCE of 2.11% under identical device configurations. PMID- 21630706 TI - On the mechanism of dimethylarginine dimethylaminohydrolase inactivation by 4 halopyridines. AB - Small molecules capable of selective covalent protein modification are of significant interest for the development of biological probes and therapeutics. We recently reported that 2-methyl-4-bromopyridine is a quiescent affinity label for the nitric oxide controlling enzyme dimethylarginine dimethylaminohydrolase (DDAH) (Johnson, C. M.; Linsky, T. W.; Yoon, D. W.; Person, M. D.; Fast, W. J. Am. Chem. Soc. 2011, 133, 1553-1562). Discovery of this novel protein modifier raised the possibility that the 4-halopyridine motif may be suitable for wider application. Therefore, the inactivation mechanism of the related compound 2 hydroxymethyl-4-chloropyridine is probed here in more detail. Solution studies support an inactivation mechanism in which the active site Asp66 residue stabilizes the pyridinium form of the inactivator, which has enhanced reactivity toward the active site Cys, resulting in covalent bond formation, loss of the halide, and irreversible inactivation. A 2.18 A resolution X-ray crystal structure of the inactivated complex elucidates the orientation of the inactivator and its covalent attachment to the active site Cys, but the structural model does not show an interaction between the inactivator and Asp66. Molecular modeling is used to investigate inactivator binding, reaction, and also a final pyridinium deprotonation step that accounts for the apparent differences between the solution-based and structural studies with respect to the role of Asp66. This work integrates multiple approaches to elucidate the inactivation mechanism of a novel 4-halopyridine "warhead," emphasizing the strategy of using pyridinium formation as a "switch" to enhance reactivity when bound to the target protein. PMID- 21630708 TI - Next-generation sequencing and its potential impact. AB - The new generation of DNA sequencing devices surpass their predecessors' outputs by many orders of magnitude, a quantitative increase with qualitative implications. Sequencing at this scale enables many advances in basic and medical research, ranging from identification of new medically relevant biological mechanisms to maturation of personalized therapies. Nevertheless, as the genomics era plainly demonstrated, massive amounts of DNA sequence information by itself cannot fully illuminate the mysteries of human physiology, including the problems that challenge predictive toxicologists. PMID- 21630709 TI - Diazaphenalenyl-containing spin sources designed by standardization of intramolecular magnetic interactions. AB - A new series of neutral radicals (DP1-DP6) based on diazaphenalenyl were designed via heteroatomic modifications. As spin sources, the designed radicals were implemented in four different diradical model systems (models I, II, III, and IV) by changing the reference radical and the linkage, and their magnetic interactions between the designed radicals and the reference radical were investigated by using density functional theory calculations. The trend in strength of magnetic interactions of radicals was found to be identical in different model systems. The strength of magnetic interactions could be understood by spin density distributions and NICS values. Our results will be helpful in designing and controlling the organic magnetic materials incorporated with diazaphenalenyl derivatives. In particular, as a new family of spin source radical, DP3 could be a potential candidate in designing new organic magnetic materials due to its strong magnetic coupling and high stability in diradical systems. PMID- 21630710 TI - Occurrence and risk assessment of four typical fluoroquinolone antibiotics in raw and treated sewage and in receiving waters in Hangzhou, China. AB - A sensitive liquid chromatography-fluorescence detection method, combined with one-step solid-phase extraction, was established for detecting the residual levels of the four typical fluoroquinolone antibiotics (ofloxacin, norfloxacin, ciprofloxacin, and enrofloxacin) in influent, effluent, and surface waters from Hangzhou, China. For the various environmental water matrices, the overall recoveries were from 76.8 to 122%, and no obvious interferences of matrix effect were observed. The limit of quantitation of this method was estimated to be 17 ng/L for ciprofloxacin and norfloxacin, 20 ng/L for ofloxacin, and 27 ng/L for enrofloxacin. All of the four typical fluoroquinolone antibiotics were found in the wastewaters and surface waters. The residual contents of the four typical fluoroquinolone antibiotics in influent, effluent, and surface water samples are 108-1405, 54-429, and 7.0-51.6 ng/L, respectively. The removal rates of the selected fluoroquinolone antibiotics were 69.5 (ofloxacin), 61.3 (norfloxacin), and 50% (enrofloxacin), indicating that activated sludge treatment is effective except for ciprofloxacin and necessary to remove these fluoroquinolone antibiotics in municipal sewage. The risk to the aquatic environment was estimated by a ratio of measured environmental concentration and predicted no effect concentration. At the concentrations, these fluoroquinolone antibiotics were found in influent, effluent, and surface waters, and they should not pose a risk for the aquatic environment. PMID- 21630711 TI - Viability of clathrate hydrates as CO2 capturing agents: a theoretical study. AB - Capture and sequestration of green house gas CO(2) is a major challenge for scientists and identifying right materials for this purpose is a task of outstanding importance. Through reliable computational studies, we have demonstrated that the clathrate cages (5(12), 4(3)5(6)6(3), 5(12)6(2), 5(12)6(4), and 5(12)6(8)) have a great potential to store CO(2). All the considered clathrates and their CO(2) inclusion complexes are optimized at B3LYP/6-31G(d) level of theory. The impact of DFT-D, M05-2X, and MP2 functionals on interaction energy were tested using various basis sets. Although different functionals and basis sets show variation in absolute IE values, the trend is consistent and does not depend on the level of the calculations. Dispersion was found important for these complexes and DFT-D shows comparable IE values with MP2 functional. The optimum and maximum cage occupancy for all the considered cages were tested on the basis of quantum chemical calculations. The maximum cage occupancy for all five considered cages (5(12), 4(3)5(6)6(3), 5(12)6(2), 5(12)6(4), and 5(12)6(8)) is one, two, two, two, and seven CO(2) molecules, respectively, and the optimum cage occupancy is one, one, one, two, and five CO(2) molecules, respectively. Thus, 5(12)6(8) cages can host up to 7 CO(2) molecules, resulting in about 32 wt %, which makes them highly promising materials. PMID- 21630712 TI - Catalytic asymmetric synthesis of a tertiary benzylic carbon center via phenol directed alkene hydrogenation. AB - An expeditious synthetic approach to chiral phenol 1, a key building block in the preparation of a series of drug candidates, is reported. The strategy includes a cost-effective and readily scalable route to cyclopentanone 3 from isobutyronitrile (10). The sterically hindered and enolizable ketone 3 was subsequently employed in a challenging Grignard addition mediated by LaCl(3).2LiCl. A novel preparation of the lanthanide reagent required for this transformation is described. To complete the process, a highly enantioselective hydrogenation step afforded the target (1). The importance of the phenol group to the success of this asymmetric transformation is discussed. PMID- 21630713 TI - Synthesis of alkylidenecyclopropanes by palladium-catalyzed reaction of propargyl substituted malonate esters with aryl halides by anti-carbopalladation pathway. AB - Palladium-catalyzed arylative cyclization of propargyl-substituted malonate esters with aryl halides offers a stereoselective approach to alkylidenecyclopropanes. The reaction proceeds by an anti-carbopalladation pathway, which guarantees the exclusive stereocontrol of the resulting double bond. The highly strained as well as densely substituted skeletons of the products facilitate further versatile transformations, which underscores the importance of the products as synthetic intermediates. PMID- 21630714 TI - Vibrational coherence from van der Waals modes in the native and molten-globule states of ZnII-substituted cytochrome c. AB - The low-frequency vibrational coherence from Zn(II)-substituted cytochrome c (ZnCytc) was characterized at room temperature in the native and acid/high-salt molten-globule states using femtosecond pump-probe, dynamic-absorption spectroscopy and impulsive excitation of the Soret absorption band. The pump probe signals observed from the native state contain two types of modulation components in the vibrational coherence. The first type is a set of slowly damped (damping time gamma > 1.5 ps) components with frequencies of 10, 30, 70, and 120 cm(-1) that are assigned to out-of-plane vibrations of the porphyrin macrocycle following similar assignments in other porphyrin systems. A similar set of components is observed in the pump-probe signal from the molten-globule state, but the signal is much less strongly modulated. The second type is a strong, very rapidly damped (gamma < 150 fs) 79 cm(-1) modulation component that is assigned to van der Waals interactions between the porphyrin and nonpolar groups in its first solvation shell from the surrounding protein structure; the line shape and intensity of this component are comparable to those observed previously for bacteriochlorophyll a and Zn(II)meso-tetrakis(N-methylpyridyl)porphyrin in solution. This component is almost completely absent from the signal from the molten-globule state. The results suggest that the van der Waals modes obtain intensity enhancement in the vibrational coherence because the attacking groups are displaced by the change of extent and/or change in shape of the pi-electron density that accompanies the pi -> pi* optical transition of the Zn(II) porphyrin. In the molten-globule state of ZnCytc, owing to the expanded hydrophobic core and to the loss of order for the groups that attack the pi electron density of the Zn(II) porphyrin, the van der Waals modes are rendered effectively inactive. These results support an assignment of the broad low frequency background in the spectrum of the vibrational coherence in purple bacterial photosynthetic reaction centers to van der Waals interactions between the primary electron donor, P, and the nonpolar protein-derived groups in its first solvation shell. PMID- 21631107 TI - Synthesis of heterocyclic homotriptycenes. AB - A series of novel heterocyclic homotriptycenes bearing furan, thiophene, and pyridine rings, 7a-f, were synthesized by intramolecular dehydration reactions of 10,10-dihetarylmethyl-9,10-dihydroanthracen-9-ols 6a-f. In the presence of acids, the secondary alcohols 6a-f show different reactions which depend on the electron densities of the attached heterocyclic rings. The initially formed carbenium ions react in an electrophilic substitution with electron-rich heterocycles. The formation of a transannular bridge (1,7-elimination) leads to homotriptycenes in high yields. When the heterocyclic ring has a moderate electron density, two competitive reactions exist, which afford 9-monosubstituted anthracenes by 1,4 elimination or 9,10-disubstituted anthracenes by a rearrangement, respectively. Electron-deficient heterocycles undergo a disproportionation to give hydrocarbons and ketones. PMID- 21631108 TI - Measurements of growth rates of an ice crystal from supercooled heavy water under microgravity conditions: basal face growth rate and tip velocity of a dendrite. AB - The growth of single ice crystals from supercooled heavy water was studied under microgravity conditions in the Japanese Experiment Module ''KIBO'' of the International Space Station (ISS). The velocities of dendrite tips parallel to the a axis and the growth rates of basal faces parallel to the c axis were both analyzed under supercooling ranging from 0.03 to 2.0 K. The velocities of dendrite tips agree with the theory for larger amounts of supercooling when the growth on the basal faces are not zero. At very low supercooling there is no growth on the basal faces. With increasing supercooling the basal faces start to grow, the growth rate changing as a function of supercooling with a power law with an exponent of about 2, with the exponent approaching 1 as supercooling increases further. We interpret the growth on the basal faces as being controlled by two-dimensional nucleation under low supercooling, with a change in the growth kinetics to spiral growth with the aid of screw dislocations with increasing supercooling then to a linear growth law. We discuss the combined effect of tip velocity and basal face kinetics on pattern formation during the growth of ice. PMID- 21631109 TI - iTRAQ-2DLC-ESI-MS/MS based identification of a new set of immunohistochemical biomarkers for classification of dysplastic nodules and small hepatocellular carcinoma. AB - The study aims to develop novel clinical immunohistochemical biomarkers for distinguishing small hepatocellular carcinoma (sHCC) from dysplastic nodules (DN). iTRAQ-2DLC-ESI-MS/MS technique was used to screen immunohistochemical biomarkers between precancerous lesions (liver cirrhosis and DN) and sHCC. A total of 1951 proteins were quantified, including 52 proteins upregulated in sHCC and 95 proteins downregulated in sHCC by at least 1.25- or 0.8-fold at p < 0.05. The selected biomarker candidates were further verified using Western blotting and immunohistochemistry. Furthermore, receiver operation characteristics (ROC) curves and logistic regression model were carried out to evaluate the diagnostic values of the biomarkers. Finally, aminoacylase-1 (ACY1) and sequestosome-1 (SQSTM1) were chosen as novel candidate biomarkers for distinction of sHCC from DN. A constructed logistic regression model included ACY1, SQSTM1, and CD34. The sensitivity and specificity of this model for distinguishing sHCC from DN was 96.1% and 96.7%. In conclusion, ACY1 and SQSTM1 were identified as novel immunohistochemical biomarkers distinguishing sHCC from DN. In conclusion, expression levels of CD34, ACY1, and SQSTM1 can be used to establish an accurate diagnostic model for distinction of sHCC from DN. PMID- 21631110 TI - Imaging intracellular pH in live cells with a genetically encoded red fluorescent protein sensor. AB - Intracellular pH affects protein structure and function, and proton gradients underlie the function of organelles such as lysosomes and mitochondria. We engineered a genetically encoded pH sensor by mutagenesis of the red fluorescent protein mKeima, providing a new tool to image intracellular pH in live cells. This sensor, named pHRed, is the first ratiometric, single-protein red fluorescent sensor of pH. Fluorescence emission of pHRed peaks at 610 nm while exhibiting dual excitation peaks at 440 and 585 nm that can be used for ratiometric imaging. The intensity ratio responds with an apparent pK(a) of 6.6 and a >10-fold dynamic range. Furthermore, pHRed has a pH-responsive fluorescence lifetime that changes by ~0.4 ns over physiological pH values and can be monitored with single-wavelength two-photon excitation. After characterizing the sensor, we tested pHRed's ability to monitor intracellular pH by imaging energy dependent changes in cytosolic and mitochondrial pH. PMID- 21631111 TI - Disruption of supported lipid bilayers by semihydrophobic nanoparticles. AB - Understanding the interaction between functional nanoparticles and cell membranes is critical to use nanomaterials for broad biomedical applications with minimal cytotoxicity. In this work, we have investigated the effect of adsorbed semihydrophobic nanoparticles (NPs) on the dynamics and morphology of model cell membranes. We have systematically varied the degree of surface hydrophobicity of carboxyl end-functionalized polystyrene NPs of varied size in buffer solutions with varied ionic strength. It is observed that semihydrophobic NPs can readily adsorb on neutral SLBs and drag lipids from SLBs to NP surfaces. Above a critical NP concentration, the disruption of SLBs is observed, accompanied with the formation and rapid growth of lipid-poor regions on NP-adsorbed SLBs. In the study of the effect of solution ionic strength on NP surface hydrophobic degree and the growth of lipid-poor regions, we have concluded that the hydrophobic interaction enhanced by screened electrostatic interaction underlies the envelopment of NPs by lipids that are attracted from SLBs to the surface of NPs or their aggregates. Hence, the formation and growth of lipid-poor regions, or vaguely referred as "pores" or "holes" in the literature, can be controlled by NP concentration, size, and surface hydrophobicity, which is critical to design functional nanomaterials for effective nanomedicine while minimizing possible cytotoxicity. PMID- 21631112 TI - Anti-inflammatory activities of mogrosides from Momordica grosvenori in murine macrophages and a murine ear edema model. AB - Momordica grosvenori (Luo Han Guo), grown primarily in Guangxi province in China, has been traditionally used for thousands of years by the Chinese to make hot drinks for the treatment of sore throat and the removal of phlegm. The natural noncaloric sweetening triterpenoid glycosides (mogrosides) contained in the M. grosvenori fruits are also antioxidative, anticarcinogenic, and helpful in preventing diabetic complications. The aim of this study was to assess the anti inflammatory properties of mogrosides in both murine macrophage RAW 264.7 cells and a murine ear edema model. The results indicate that mogrosides can inhibit inflammation induced by lipopolysaccharides (LPS) in RAW 264.7 cells by down regulating the expression of key inflammatory genes iNOS, COX-2, and IL-6 and up regulating some inflammation protective genes such as PARP1, BCL2l1, TRP53, and MAPK9. Similarly, in the murine ear edema model, 12-O-tetradecanoylphorbol-13 acetate-induced inflammation was inhibited by mogrosides by down-regulating COX-2 and IL-6 and up-regulating PARP1, BCL2l1, TRP53, MAPK9, and PPARdelta gene expression. This study shows that the anticancer and antidiabetic effects of M. grosvenori may result in part from its anti-inflammatory activity. PMID- 21631113 TI - Impact of low hydration of barley grain on beta-glucan degradation and lipid transfer protein (LTP1) modifications during the malting process. AB - One of the objectives of the malting industry is to reduce the energy cost during kilning without major effect on malt quality. In this study, the impact of a low hydration steeping process on lipid transfer protein (LTP1) modifications and beta-glucan breakdown was evaluated in low (LH) and high (HH) hydrated malts. LTP1 modifications analyzed by MS/MS revealed acylation, glycation, and disulfide bond breakage in both LH and HH malts. LTP1 free amine content measurement and fluorescence of Maillard protein adducts revealed no significant difference between LH and HH malts. Immunolabeling of LTP1 during malting highlighted the diffusion of the protein from the aleurone layer to the endosperm at the end of steeping in both LH and HH malts. By contrast, a significant higher amount of beta-glucans was measured in LH malts after five days of germination, whereas no significant difference between LH and HH malts was revealed through immunostaining of beta-glucans or evaluation of the endosperm integrity after seven days of germination. The possibility to reduce the effects of a low hydration steeping process on beta-glucan hydrolysis by increasing germination time was discussed. PMID- 21631115 TI - Clustering in complex fluids. PMID- 21631116 TI - A unique paradigm for a Turn-ON near-infrared cyanine-based probe: noninvasive intravital optical imaging of hydrogen peroxide. AB - The development of highly sensitive fluorescent probes in combination with innovative optical techniques is a promising strategy for intravital noninvasive quantitative imaging. Cyanine fluorochromes belong to a superfamily of dyes that have attracted substantial attention in probe design for molecular imaging. We have developed a novel paradigm to introduce a Turn-ON mechanism in cyanine molecules, based on a distinctive change in their pi-electrons system. Our new cyanine fluorochrome is synthesized through a simple two-step procedure and has an unprecedented high fluorescence quantum yield of 16% and large extinction coefficient of 52,000 M(-1)cm(-1). The synthetic strategy allows one to prepare probes for various analytes by introducing a specific triggering group on the probe molecule. The probe was equipped with a corresponding trigger and demonstrated efficient imaging of endogenous hydrogen peroxide, produced in an acute lipopolysaccharide-induced inflammation model in mice. This approach provides, for the first time, an available methodology to prepare modular molecular Turn-ON probes that can release an active cyanine fluorophore upon reaction with specific analyte. PMID- 21631117 TI - Formation of organo-highly charged mica. AB - The interlayer space of the highly charged synthetic Na-Mica-4 can be modified by ion-exchange reactions involving the exchange of inorganic Na(+) cations by surfactant molecules, which results in the formation of an organophilic interlayer space. The swelling and structural properties of this highly charged mica upon intercalation with n-alkylammonium (RNH(3))(+) cations with varying alkyl chain lengths (R = C12, C14, C16, and C18) have been reported. The stability, fine structure, and evolution of gaseous species from alkylammonium Mica-4 are investigated in detail by conventional thermogravimetric analysis (TGA), Fourier transform infrared spectroscopy (FTIR), in situ X-ray diffraction (XRD), and solid-state nuclear magnetic resonance (MAS NMR) techniques. The results clearly show the total adsorption of n-alkylammonium cations in the interlayer space which expands as needed to accommodate intercalated surfactants. The surfactant packing is quite ordered at room temperature, mainly involving a paraffin-type bilayer with an all-trans conformation, in agreement with the high density of the organic compounds in the interlayer space. At temperatures above 160 degrees C, the surfactant molecules undergo a transformation that leads to a liquid-like conformation, which results in a more disordered phase and expansion of the interlayer space. PMID- 21631118 TI - Aliphatic ketones from Ruta chalepensis (Rutaceae) induce paralysis on root knot nematodes. AB - This paper reports on the use of Ruta chalepensis L. extracts as a potential nematicide against root knot nematodes Meloidogyne incognita and Meloidogyne javanica . The essential oil (REO) and methanol extract (RME) of R. chalepensis were tested against second-stage juveniles, with REO inducing paralysis in both species (EC(50/1d) = 77.5 and 107.3 mg/L) and RME being selective for M. incognita (EC(50/1d) = 1001 mg/L). Chemical characterization of extracts was done by means of GC-MS and LC-MS, revealing mainly aliphatic ketones and coumarins, respectively. The first-ranking volatile nematicidal component in terms of individual activity against both species was 2-undecanone (EC(50) = 20.6 and 22.5 mg/L for M. incognita and M. javanica, respectively). This fact together with its high concentration in the most active extract found in this study, namely, REO (2926 mg/kg), categorizes 2-undecanone among the nematicidal principles of R. chalepensis. On the contrary, coumarins rutin and 8-methoxypsoralen were not found to be nematicidal at concentrations of <=500 mg/L. Interestingly, M. incognita was found more sensitive than M. javanica. PMID- 21631119 TI - Isolation and characterization of three novel peptides from casein hydrolysates that stimulate the growth of mixed cultures of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus. AB - In this study, sodium caseinate hydrolysates produced by papain with strong growth-stimulating activity for Streptococcus thermophilus (St) and Lactobacillus delbrueckii subsp. bulgaricus (Lb) were obtained. A series of separation methods including ultrafiltration, macroporous adsorption resin chromatography, gel filtration chromatography, and reverse-phase high-performance liquid chromatography (RP-HPLC) were applied to isolate and purify the peptide(s), which were mainly responsible for the activity. Finally, three novel growth-stimulating peptides, H-2-A, F2-c, and F2-b, corresponding to amino acid residues 29-35 and 103-108 of bovine alpha(S2)-casein and 181-186 of bovine alpha(S1)-casein, respectively, were obtained. With supplementation of H-2-A, F2-b, or F2-c at a protein concentration of 0.3%, the biomass yield of these two lactic acid bacteria (LAB) was enhanced by 193.3, 166.7, or 151.7%, respectively. In addition, there were significant (p < 0.05) increases in viable counts of St and lactic acid production of LAB in the presence of the purified peptides. PMID- 21631120 TI - Characterization of primary amine capped CdSe, ZnSe, and ZnS quantum dots by FT IR: determination of surface bonding interaction and identification of selective desorption. AB - Surface ligands of semiconductor quantum dots (QDs) critically influence their properties and functionalities. It is of strong interest to understand the structural characteristics of surface ligands and how they interact with the QDs. Three quantum dot (QD) systems (CdSe, ZnSe, and ZnS) with primary aliphatic amine capping ligands were characterized primarily by FT-IR spectroscopy as well as NMR, UV-vis, and fluorescence spectroscopy, and by transmission electron microscopy (TEM). Representative primary amines ranging from 8 to 16 carbons were examined in the vapor phase, KBr pellet, and neat and were compared to the QD samples. The strongest hydrogen-bonding effects of the adsorbed ligands were observed in CdSe QDs with the weakest observed in ZnS QDs. There was an observed splitting of the N-H scissoring mode from 1610 cm(-1) in the neat sample to 1544 and 1635 cm(-1) when bound to CdSe QDs, which had the largest splitting of this type. The splitting is attributed to amine ligands bound to either Cd or Se surface sites, respectively. The effect of exposure of the QDs dispersed in nonpolar medium to methanol as a crashing agent was also examined. In the CdSe system, the Cd-bound scissoring mode disappeared, possibly due to methanol replacing surface cadmium sites. The opposite was observed for ZnSe QDs, in which the Se-bound scissoring mode disappeared. It was concluded that surface coverage and ligand bonding partners could be characterized by FT-IR and that selective removal of surface ligands could be achieved through introduction of competitive binding interactions at the surface. PMID- 21631121 TI - High-fidelity solvent-resistant replica molding of hydrophobic polymer surfaces produced by femtosecond laser nanofabrication. AB - We demonstrate that hydrophobic areas formed by femtosecond laser irradiation on poly(methyl methacrylate) (PMMA) and polystyrene (PS) polymer substrates can be faithfully replicated on samples of the same material via a solvent-resistant perfluoropolyether (PFPE) elastomer mold. The replicated PMMA and PS samples show nearly identical micro-nanoscale topography and hydrophobic wetting characteristics as the laser-patterned master substrates. This work combines the femtosecond laser capability of spatially tailoring the wettability with a high resolution parallel replication method, offering the potential for the efficient production of microfluidic devices with selectively tailored flow behavior. PMID- 21631122 TI - Liquid marbles prepared from pH-responsive sterically stabilized latex particles. AB - Submicrometer-sized pH-responsive sterically stabilized polystyrene (PS) latex particles were synthesized by dispersion polymerization in isopropyl alcohol with a poly[2-(diethylamino)ethyl methacrylate]- (PDEA-) based macroinitiator. These PDEA-PS latexes were extensively characterized with respect to their particle size distribution, morphology, chemical composition, and pH-responsive behavior. Millimeter- and centimeter-sized "liquid marbles" with aqueous volumes varying between 15 MUL and 2.0 mL were readily prepared by rolling water droplets on the dried PDEA-PS latex powder. The larger liquid marbles adopted nonspherical shapes due to gravitational forces; analysis of this deformation enabled the surface tension to be estimated. Scanning electron microscopy and fluorescence microscopy studies indicated that flocs of the PDEA-PS particles were adsorbed at the surface of these water droplets, leading to stable liquid marbles. The relative mechanical integrity of the liquid marbles prepared from alkaline aqueous solution (pH 10) was higher than those prepared from acidic aqueous solution (pH 2) as judged by droplet roller experiments. These liquid marbles exhibited long term stability (over 1 h) when transferred onto the surface of liquid water, provided that the solution pH of the subphase was above pH 8. In contrast, the use of acidic solutions led to immediate disintegration of these liquid marbles within 10 min, with dispersal of the PDEA-PS latex particles in the aqueous solution. Thus the critical minimum solution pH required for long-term liquid marble stability correlates closely with the known pK(a) value of 7.3 for the PDEA stabilizer chains. Stable liquid marbles were also successfully prepared from aqueous Gellan gum solution and glycerol. PMID- 21631123 TI - Perfluorocarbon thin films and polymer brushes on stainless steel 316 L for the control of interfacial properties. AB - Perfluorocarbon thin films and polymer brushes were formed on stainless steel 316 L (SS316L) to control the surface properties of the metal oxide. Substrates modified with the films were characterized using diffuse reflectance infrared Fourier transform spectroscopy (DRIFT), contact angle analysis, atomic force microscopy (AFM), and cyclic voltammetry (CV). Perfluorooctadecanoic acid (PFOA) was used to form thin films by self-assembly on the surface of SS316L. Polypentafluorostyrene (PFS) polymer brushes were formed by surface-initiated polymerization using SAMs of 16-phosphonohexadecanoic acid (COOH-PA) as the base. PFOA and PFS were effective in significantly reducing the surface energy and thus the interfacial wetting properties of SS316L. The SS316L control exhibited a surface energy of 38 mN/m compared to PFOA and PFS modifications, which had surface energies of 22 and 24 mN/m, respectively. PFOA thin films were more effective in reducing the surface energy of the SS316L compared to PFS polymer brushes. This is attributed to the ordered PFOA film presenting aligned CF(3) terminal groups. However, PFS polymer brushes were more effective in providing corrosion protection. These low-energy surfaces could be used to provide a hydrophobic barrier that inhibits the corrosion of the SS316L metal oxide surface. PMID- 21631124 TI - Isotopic hydration of cellobiose: vibrational spectroscopy and dynamical simulations. AB - The conformation and structural dynamics of cellobiose, one of the fundamental building blocks in nature, its C4' epimer, lactose, and their microhydrated complexes, isolated in the gas phase, have been explored through a combination of experiment and theory. Their structures at low temperature have been determined through double resonance, IR-UV vibrational spectroscopy conducted under molecular beam conditions, substituting D(2)O for H(2)O to separate isotopically, the carbohydrate (OH) bands from the hydration (OD) bands. Car-Parrinello (CP2K) simulations, employing dispersion corrected density functional potentials and conducted "on-the-fly" from ~20 to ~300 K, have been used to explore the consequences of raising the temperature. Comparisons between the experimental data, anharmonic vibrational self-consistent field calculations based upon ab initio potentials, and the CP2K simulations have established the role of anharmonicity; the reliability of classical molecular dynamics predictions of the vibrational spectra of carbohydrates and the accuracy of the dispersion corrected (BLYP-D) force fields employed; the structural consequences of increasing hydration; and the dynamical consequences of increasing temperature. The isolated and hydrated cellobiose and lactose units both present remarkably rigid structures: their glycosidic linkages adopt a "cis" (anti-phi and syn-psi) conformation bound by inter-ring hydrogen bonds. This conformation is maintained when the temperature is increased to ~300 K and it continues to be maintained when the cellobiose (or lactose) unit is hydrated by one or two explicitly bound water molecules. Despite individual fluctuations in the intra- and intermolecular hydrogen bonding pattern and some local structural motions, the water molecules remain locally bound and the isolated carbohydrates remain trapped within the cis potential well. The Car-Parrinello dynamical simulations do not suggest any accessible pathway to the trans conformations that are formed in aqueous solution and are widespread in nature. PMID- 21631125 TI - The methylation effect in medicinal chemistry. PMID- 21631126 TI - Mathematical Biosciences and Engineering. From the guest editors. PMID- 21631127 TI - Investigating the steady state of multicellular spheroids by revisiting the two fluid model. AB - In this paper we examine the steady state of tumour spheroids considering a structure in which the central necrotic region contains an inner liquid core surrounded by dead cells that keep some mechanical integrity. This partition is a consequence of assuming that a finite delay is required for the degradation of dead cells into liquid. The phenomenological assumption of constant local volume fraction of cells is also made. The above structure is coupled with a simple mechanical model that views the cell component as a viscous fluid and the extracellular liquid as an inviscid fluid. By imposing the continuity of the normal stress throughout the whole spheroid, we show that a steady state can exist only if the forces on cells at the outer boundary (provided e.g. by a surface tension) are intense enough, and in such a case we can compute the stationary radius. By giving reasonable values to the parameters, the model predicts that the stationary radius decreases with the external oxygen concentration, as expected from experimental observations. PMID- 21631129 TI - Modeling and simulation of some cell dispersion problems by a nonparametric method. AB - Starting from the classical descriptions of cell motion we propose some ways to enhance the realism of modeling and to account for interesting features like allowing for a random switching between biased and unbiased motion or avoiding a set of obstacles. For this complex behavior of the cell population we propose new models and also provide a way to numerically assess the macroscopic densities of interest upon using a nonparametric estimation technique. Up to our knowledge, this is the only method able to numerically handle the entire complexity of such settings. PMID- 21631128 TI - A mathematical model for chronic wounds. AB - Chronic wounds are often associated with ischemic conditions whereby the blood vascular system is damaged. A mathematical model which accounts for these conditions is developed and computational results are described in the two dimensional radially symmetric case. Preliminary results for the three dimensional axially symmetric case are also included. PMID- 21631130 TI - Questions from the fourth son: a clinician reflects on immunomonitoring, surrogate markers and systems biology. AB - The fourth son is the one who doesn't even know how to ask a question. Tumor immunology is challenged by the failure to identify reliable surrogate markers in vaccine and other experimental therapies for cancer; perhaps investigators haven't yet asked the right questions. Unlike prophylactic vaccines for infectious disease, where the development of antibody is considered a satisfactory endpoint, no such endpoint exists for human therapeutic vaccines. Why is this? Despite an extensive roster of in vitro assays that correlate immune responses to favorable clinical outcomes, no assay is sufficiently reliable to be usefully predictive for vaccine therapy. The discussion reviews some of the historical developments in tumor immunology and the problem of defining a causal relationship when strong correlations are identified. The development of mathematical models from empirical data may help inform the clinician/scientist about underlying mechanisms and help frame new testable hypotheses. PMID- 21631131 TI - Mathematical modeling of cyclic treatments of chronic myeloid leukemia. AB - Cyclic treatment strategies in Chronic Myeloid Leukemia (CML) are characterized by alternating applications of two (or more) different drugs, given one at a time. One of the main causes for treatment failure in CML is the generation of drug resistance by mutations of cancerous cells. We use mathematical methods to develop general guidelines on optimal cyclic treatment scheduling, with the aim of minimizing the resistance generation. We define a condition on the drugs' potencies which allows for a relatively successful application of cyclic therapies. We find that the best strategy is to start with the stronger drug, but use longer cycle durations for the weaker drug. We further investigate the situation where a degree of cross-resistance is present, such that certain mutations cause cells to become resistant to both drugs simultaneously. PMID- 21631132 TI - Optimal and suboptimal protocols for a mathematical model for tumor anti angiogenesis in combination with chemotherapy. AB - We consider the problem of minimizing the tumor volume with a priori given amounts of anti-angiogenic and cytotoxic agents. For one underlying mathematical model, optimal and suboptimal solutions are given for four versions of this problem: the case when only anti-angiogenic agents are administered, combination treatment with a cytotoxic agent, and when a standard linear pharmacokinetic equation for the anti-angiogenic agent is added to each of these models. It is shown that the solutions to the more complex models naturally build upon the simplified versions. This gives credence to a modeling approach that starts with the analysis of simplified models and then adds increasingly more complex and medically relevant features. Furthermore, for each of the problem formulations considered here, there exist excellent simple piecewise constant controls with a small number of switchings that virtually replicate the optimal values for the objective. PMID- 21631133 TI - A model of competing saturable kinetic processes with application to the pharmacokinetics of the anticancer drug paclitaxel. AB - A saturable multi-compartment pharmacokinetic model for the anti-cancer drug paclitaxel is proposed based on a meta-analysis of pharmacokinetic data published over the last two decades. We present and classify the results of time series for the drug concentration in the body to uncover the underlying power laws. Two dominant fractional power law exponents were found to characterize the tails of paclitaxel concentration-time curves. Short infusion times led to a power exponent of -1.57 +/- 0.14, while long infusion times resulted in tails with roughly twice the exponent. Curves following intermediate infusion times were characterized by two power laws. An initial segment with the larger slope was followed by a long-time tail characterized by the smaller exponent. The area under the curve and the maximum concentration exhibited a power law dependence on dose, both with compatible fractional power exponents. Computer simulations using the proposed model revealed that a two-compartment model with both saturable distribution and elimination can reproduce both the single and crossover power laws. Also, the nonlinear dose-dependence is correlated with the empirical power law tails. The longer the infusion time the better the drug delivery to the tumor compartment is a clinical recommendation we propose. PMID- 21631134 TI - Robustness of optimal controls for a class of mathematical models for tumor anti angiogenesis. AB - We describe optimal protocols for a class of mathematical models for tumor anti angiogenesis for the problem of minimizing the tumor volume with an a priori given amount of vessel disruptive agents. The family of models is based on a biologically validated model by Hahnfeldt et al. and includes a modification by Ergun et al, but also provides two new variations that interpolate the dynamics for the vascular support between these existing models. The biological reasoning for the modifications of the models will be presented and we will show that despite quite different modeling assumptions, the qualitative structure of optimal controls is robust. For all the systems in the class of models considered here, an optimal singular arc is the defining element and all the syntheses of optimal controlled trajectories are qualitatively equivalent with quantitative differences easily computed. PMID- 21631135 TI - Tumor cells proliferation and migration under the influence of their microenvironment. AB - It is well known that tumor microenvironment affects tumor growth and metastasis: Tumor cells may proliferate at different rates and migrate in different patterns depending on the microenvironment in which they are embedded. There is a huge literature that deals with mathematical models of tumor growth and proliferation, in both the avascular and vascular phases. In particular, a review of the literature of avascular tumor growth (up to 2006) can be found in Lolas (G. Lolas, Lecture Notes in Mathematics, Springer Berlin / Heidelberg, 1872, 77 (2006)). In this article we report on some of our recent work. We consider two aspects, proliferation and of migration, and describe mathematical models based on in vitro experiments. Simulations of the models are in agreement with experimental results. The models can be used to generate hypotheses regarding the development of drugs which will confine tumor growth. PMID- 21631136 TI - Periodically forced discrete-time SIS epidemic model with disease induced mortality. AB - We use a periodically forced SIS epidemic model with disease induced mortality to study the combined effects of seasonal trends and death on the extinction and persistence of discretely reproducing populations. We introduce the epidemic threshold parameter, R0 , for predicting disease dynamics in periodic environments. Typically, R0 <1 implies disease extinction. However, in the presence of disease induced mortality, we extend the results of Franke and Yakubu to periodic environments and show that a small number of infectives can drive an otherwise persistent population with R0 >1 to extinction. Furthermore, we obtain conditions for the persistence of the total population. In addition, we use the Beverton-Holt recruitment function to show that the infective population exhibits period-doubling bifurcations route to chaos where the disease-free susceptible population lives on a 2-cycle (non-chaotic) attractor. PMID- 21631137 TI - Sensitivity of hemodynamics in a patient specific cerebral aneurysm to vascular geometry and blood rheology. AB - Newtonian and generalized Newtonian mathematical models for blood flow are compared in two different reconstructions of an anatomically realistic geometry of a saccular aneurysm, obtained from rotational CTA and differing to within image resolution. The sensitivity of the flow field is sought with respect to geometry reconstruction procedure and mathematical model choice in numerical simulations. Taking as example a patient specific intracranial aneurysm located on an outer bend under steady state simulations, it is found that the sensitivity to geometry variability is greater, but comparable, to the one of the rheological model. These sensitivities are not quantifiable a priori. The flow field exhibits a wide range of shear stresses and slow recirculation regions that emphasize the need for careful choice of constitutive models for the blood. On the other hand, the complex geometrical shape of the vessels is found to be sensitive to small scale perturbations within medical imaging resolution. The sensitivity to mathematical modeling and geometry definition are important when performing numerical simulations from in vivo data, and should be taken into account when discussing patient specific studies since differences in wall shear stress range from 3% to 18%. PMID- 21631138 TI - Blood coagulation dynamics: mathematical modeling and stability results. AB - The hemostatic system is a highly complex multicomponent biosystem that under normal physiologic conditions maintains the fluidity of blood. Coagulation is initiated in response to endothelial surface vascular injury or certain biochemical stimuli, by the exposure of plasma to Tissue Factor (TF), that activates platelets and the coagulation cascade, inducing clot formation, growth and lysis. In recent years considerable advances have contributed to understand this highly complex process and some mathematical and numerical models have been developed. However, mathematical models that are both rigorous and comprehensive in terms of meaningful experimental data, are not available yet. In this paper a mathematical model of coagulation and fibrinolysis in flowing blood that integrates biochemical, physiologic and rheological factors, is revisited. Three dimensional numerical simulations are performed in an idealized stenosed blood vessel where clot formation and growth are initialized through appropriate boundary conditions on a prescribed region of the vessel wall. Stability results are obtained for a simplified version of the clot model in quiescent plasma, involving some of the most relevant enzymatic reactions that follow Michaelis Menten kinetics, and having a continuum of equilibria. PMID- 21631139 TI - Regulation of modular Cyclin and CDK feedback loops by an E2F transcription oscillator in the mammalian cell cycle. AB - The cell cycle is regulated by a large number of enzymes and transcription factors. We have developed a modular description of the cell cycle, based on a set of interleaved modular feedback loops, each leading to a cyclic behavior. The slowest loop is the E2F transcription and ubiquitination, which determines the cycling frequency of the entire cell cycle. Faster feedback loops describe the dynamics of each Cyclin by itself. Our model shows that the cell cycle progression as well as the checkpoints of the cell cycle can be understood through the interactions between the main E2F feedback loop and the driven Cyclin feedback loops. Multiple models were proposed for the cell cycle dynamics; each with differing basic mechanisms. We here propose a new generic formalism. In contrast with existing models, the proposed formalism allows a straightforward analysis and understanding of the dynamics, neglecting the details of each interaction. This model is not sensitive to small changes in the parameters used and it reproduces the observed behavior of the transcription factor E2F and different Cyclins in continuous or regulated cycling conditions. The modular description of the cell cycle resolves the gap between cyclic models, solely based on protein-protein reactions and transcription reactions based models. Beyond the explanation of existing observations, this model suggests the existence of unknown interactions, such as the need for a functional interaction between Cyclin B and retinoblastoma protein (Rb) de-phosphorylation. PMID- 21631140 TI - Preliminary analysis of an agent-based model for a tick-borne disease. AB - Ticks have a unique life history including a distinct set of life stages and a single blood meal per life stage. This makes tick-host interactions more complex from a mathematical perspective. In addition, any model of these interactions must involve a significant degree of stochasticity on the individual tick level. In an attempt to quantify these relationships, I have developed an individual based model of the interactions between ticks and their hosts as well as the transmission of tick-borne disease between the two populations. The results from this model are compared with those from previously published differential equation based population models. The findings show that the agent-based model produces significantly lower prevalence of disease in both the ticks and their hosts than what is predicted by a similar differential equation model. PMID- 21631141 TI - Bacteria-phagocyte dynamics, axiomatic modelling and mass-action kinetics. AB - Axiomatic modeling is ensued to provide a family of models that describe bacterial growth in the presence of phagocytes, or, more generally, prey dynamics in a large spatially homogenous eco-system. A classification of the possible bifurcation diagrams that arise in such models is presented. It is shown that other commonly used models that do not belong to this class may miss important features that are associated with the limited growth curve of the bacteria (prey) and the saturation associated with the phagocytosis (predator kill) term. Notably, these features appear at relatively low concentrations, much below the saturation range. Finally, combining this model with a model of neutrophil dynamics in the blood after chemotherapy treatments we obtain new insights regarding the development of infections under neutropenic conditions. PMID- 21631142 TI - Physiologically structured populations with diffusion and dynamic boundary conditions. AB - We consider a linear size-structured population model with diffusion in the size space. Individuals are recruited into the population at arbitrary sizes. We equip the model with generalized Wentzell-Robin (or dynamic) boundary conditions. This approach allows the modelling of populations in which individuals may have distinguished physiological states. We establish existence and positivity of solutions by showing that solutions are governed by a positive quasicontractive semigroup of linear operators on the biologically relevant state space. These results are obtained by establishing dissipativity of a suitably perturbed semigroup generator. We also show that solutions of the model exhibit balanced exponential growth, that is, our model admits a finite-dimensional global attractor. In case of strictly positive fertility we are able to establish that solutions in fact exhibit asynchronous exponential growth. PMID- 21631143 TI - Adaptive response and enlargement of dynamic range. AB - Many membrane channels and receptors exhibit adaptive, or desensitized, response to a strong sustained input stimulus, often supported by protein activity dependent inactivation. Adaptive response is thought to be related to various cellular functions such as homeostasis and enlargement of dynamic range by background compensation. Here we study the quantitative relation between adaptive response and background compensation within a modeling framework. We show that any particular type of adaptive response is neither sufficient nor necessary for adaptive enlargement of dynamic range. In particular a precise adaptive response, where system activity is maintained at a constant level at steady state, does not ensure a large dynamic range neither in input signal nor in system output. A general mechanism for input dynamic range enlargement can come about from the activity-dependent modulation of protein responsiveness by multiple biochemical modification, regardless of the type of adaptive response it induces. Therefore hierarchical biochemical processes such as methylation and phosphorylation are natural candidates to induce this property in signaling systems. PMID- 21631144 TI - Use of quasi-normal form to examine stability of tumor-free equilibrium in a mathematical model of BCG treatment of bladder cancer. AB - Understanding the dynamics of human hosts and tumors is of critical importance. A mathematical model was developed by Bunimovich-Mendrazitsky et al., who explored the immune response in bladder cancer as an effect of BCG treatment. This treatment exploits the host's own immune system to boost a response that will enable the host to rid itself of the tumor. Although this model was extensively studied using numerical simulation, no analytical results on global tumor dynamics were originally presented. In this work, we analyze stability in a mathematical model for BCG treatment of bladder cancer based on the use of quasi normal form and stability theory. These tools are employed in the critical cases, especially when analysis of the linearized system is insufficient. Our goal is to gain a deeper insight into the BCG treatment of bladder cancer, which is based on a mathematical model and biological considerations, and thereby to bring us one step closer to the design of a relevant clinical protocol. PMID- 21631145 TI - Glucose level regulation via integral high-order sliding modes. AB - Diabetes is a condition in which the body either does not produce enough insulin, or does not properly respond to it. This causes the glucose level in blood to increase. An algorithm based on Integral High-Order Sliding Mode technique is proposed, which keeps the normal blood glucose level automatically releasing insulin into the blood. The system is highly insensitive to inevitable parametric and model uncertainties, measurement noises and small delays. PMID- 21631146 TI - Modelling seasonal influenza in Israel. AB - Mathematical modeling approaches are used to study the epidemic dynamics of seasonal influenza in Israel. The recent availability of highly resolved ten year timeseries of influenza cases provides an opportunity for modeling and estimating important epidemiological parameters in the Israeli population. A simple but well known SIR discrete-time deterministic model was fitted to consecutive epidemics allowing estimation of the initial number of susceptibles in the population S0, as well as the reproductive number R0 each year. The results were corroborated by implementing a stochastic model and using a maximum likelihood approach. The paper discusses the difficulties in estimating these important parameters especially when the reporting rate of influenza cases might only be known with limited accuracy, as is generally the case. In such situations invariant parameters such as the percentage of susceptibles infected, and the effective reproductive rate might be preferred, as they do not depend on reporting rate. Results are given based on the Israeli timeseries. PMID- 21631147 TI - On some models for cancer cell migration through tissue networks. AB - We propose some models allowing to account for relevant processes at the various scales of cancer cell migration through tissue, ranging from the receptor dynamics on the cell surface over degradation of tissue fibers by protease and soluble ligand production towards the behavior of the entire cell population. For a genuinely mesoscopic version of these models we also provide a result on the local existence and uniqueness of a solution for all biologically relevant space dimensions. PMID- 21631148 TI - New approach to modeling of antiangiogenic treatment on the basis of Hahnfeldt et al. model. AB - In the paper we propose a new methodology in modeling of antiangiogenic treatment on the basis of well recognized model formulated by Hahnfeldt et al. in 1999. On the basis of the Hahnfeldt et al. model, with the usage of the optimal control theory, some protocols of antiangiogenic treatment were proposed. However, in our opinion the formulation of that model is valid only for the antivascular treatment, that is treatment that is focused on destroying endothelial cells. Therefore, we propose a modification of the original model which is valid in the case of the antiangiogenic treatment, that is treatment which is focused on blocking angiogenic signaling. We analyze basic mathematical properties of the proposed model and present some numerical simulations. PMID- 21631150 TI - A delay-differential equation model of HIV related cancer-immune system dynamics. AB - In the human body, the appearance of tumor cells usually turns on the defensive immune mechanisms. It is therefore of great importance to understand links between HIV related immunosuppression and cancer prognosis. In the paper we present a simple model of HIV related cancer - immune system interactions in vivo which takes into account a delay describing the time needed by CD T lymphocyte to regenerate after eliminating a cancer cell. The model assumes also the linear response of immune system to tumor presence. We perform a mathematical analysis of the steady states stability and discuss the biological meanings of these steady states. Numerical simulations are also presented to illustrate the predictions of the model. PMID- 21631149 TI - Persistence and emergence of X4 virus in HIV infection. AB - Approximately 50% of late-stage HIV patients develop CXCR4-tropic (X4) virus in addition to CCR5-tropic (R5) virus. X4 emergence occurs with a sharp decline in CD4+ T cell counts and accelerated time to AIDS. Why this phenotypic switch to X4 occurs is not well understood. Previously, we used numerical simulations of a mathematical model to show that across much of parameter space a promising new class of antiretroviral treatments, CCR5 inhibitors, can accelerate X4 emergence and immunodeficiency. Here, we show that mathematical model to be a minimal activation-based HIV model that produces a spontaneous switch to X4 virus at a clinically-representative time point, while also matching in vivo data showing X4 and R5 coexisting and competing to infect memory CD4+ T cells. Our analysis shows that X4 avoids competitive exclusion from an initially fitter R5 virus due to X4v unique ability to productively infect nave CD4+ T cells. We further justify the generalized conditions under which this minimal model holds, implying that a phenotypic switch can even occur when the fraction of activated nave CD4+ T cells increases at a slower rate than the fraction of activated memory CD4+ T cells. We find that it is the ratio of the fractions of activated nave and memory CD4+ T cells that must increase above a threshold to produce a switch. This occurs as the concentration of CD4+ T cells drops beneath a threshold. Thus, highly active antiretroviral therapy (HAART), which increases CD4+ T cell counts and decreases cellular activation levels, inhibits X4 viral growth. However, we show here that even in the simplest dual-strain framework, competition between R5 and X4 viruses often results in accelerated X4 emergence in response to CCR5 inhibition, further highlighting the potential danger of anti-CCR5 monotherapy in multi-strain HIV infection. PMID- 21631151 TI - A multiscale model of the bone marrow and hematopoiesis. AB - The bone marrow is necessary for renewal of all hematopoietic cells and critical for maintenance of a wide range of physiologic functions. Multiple human diseases result from bone marrow dysfunction. It is also the site in which liquid tumors, including leukemia and multiple myeloma, develop as well as a frequent site of metastases. Understanding the complex cellular and microenvironmental interactions that govern normal bone marrow function as well as diseases and cancers of the bone marrow would be a valuable medical advance. Our goal is the development of a spatially-explicit in silico model of the bone marrow to understand both its normal function and the evolutionary dynamics that govern the emergence of bone marrow malignancy. Here we introduce a multiscale computational model of the bone marrow that incorporates three distinct spatial scales, cell, hematopoietic subunit, whole marrow. Our results, using parameter estimates from literature, recapitulates normal bone marrow function and suggest an explanation for the fractal-like structure of trabeculae and sinuses in the marrow, which would be an optimization of the hematopoietic function in order to maximize the number of mature blood cells produced daily within the volumetric restrictions of the marrow. PMID- 21631152 TI - Use of health care in the main area of Sami habitation in Norway - catching up with national expenditure rates. AB - INTRODUCTION: For many years political and professional concerns have centred on the health service access of Norway's modern Indigenous Sami people. Thirty years ago, a study determined that a low rate of health expenditure on Sami patients had lead to inferior health services for the Sami people, with their average consultation rate 6 times lower than the Norwegian national average. Since 1980, there have been few studies of differences in the utilization of medical services between the Sami people and the rest of the Norwegian population. There are few official statistics relating to the ethnic category Sami. This study explored the present utilization of healthcare services among the Sami people by investigating Sami municipalities' current expenditure on somatic hospital and specialist service. METHODS: To assess the use of health care in Sami municipalities, data on expenditure of somatic hospitals and specialist services were retrieved from the Norwegian Patient Registry, and age- and sex-adjusted expenditure rates were calculated. Predominantly Sami and non-Sami municipalities were compared, as well as a comparison with the national average. Factors considered to be explanatory variables for expenditure rates were distance to care, the supply and characteristics of the healthcare system, and the stability of GPs. RESULTS: The overall public hospital expenditure in Sami municipalities was above the national average and equivalent to corresponding municipalities in the same geographical area. However, there was considerable variation among the Sami municipalities. The age groups 35-49 and 50-64 years in all Sami municipalities had higher expenditure rates than the national average regarding out-patient contacts and hospitalizations, while the expenditure on the elderly (&#8805;80 years) was below the national average in most Sami municipalities. In addition to the public sector, there was a considerable volume of private practice specialist health care, mostly public funded and in urban parts of Norway. If the use of specialists in private practice is included, there is less variation in total out patient expenditure rates in the Sami municipalities, with one exception. The municipalities with the lowest rate of public expenditure have the highest rate of private expenditure. CONCLUSION: No marked differences in healthcare expenditure was observed between the Sami and other municipalities. Overall healthcare use in Sami municipalities is above the national average and similar to corresponding municipalities in the same geographic area. However, a considerable variation in expenditure was observed among the Sami municipalities. These results do not indicate that ethnic barriers prevent Sami inhabitants from utilization of somatic hospital and specialist services. Disregarding the magnitude of expenditure, however, it is not possible to exclude that Sami patients experience a patient-physician relationship of lower quality. PMID- 21631153 TI - Residual undifferentiated cells during differentiation of induced pluripotent stem cells in vitro and in vivo. AB - Induced pluripotent stem (iPS) cells are a potential cell source for regenerative medicine. However, the tumorigenicity of iPS cells is a big concern for clinical application. In addition to the genetic manipulation of the reprogramming process and the greater risk of tumor formation, it is unclear whether iPS cells with normal development potential are still tumorigenic. Here, we investigated 3 mouse iPS cell lines, including one line that is able to generate full-term mice via tetraploid blastocyst complementation. We found that a small number of undifferentiated iPS cells could be steadily isolated and expanded after long term differentiation of cells in vitro or in vivo. The residual undifferentiated iPS cells could be expanded and redifferentiated, and undifferentiated pluripotent stem cells could again be isolated after further rounds of differentiation, suggesting that residual undifferentiated iPS cells could not be eliminated by extended cell differentiation. The residual undifferentiated cells could form teratomas in vivo, indicating that they are a potential tumorigenic risk during transplantation. These findings prompt us to reconsider the strategies for solving the tumorigenic problem of iPS cells, not only focusing on improving the reprogramming process. PMID- 21631154 TI - Induction of Wnt/beta-catenin signaling in mouse mesenchymal stem cells is associated with activation of the p130 and E2f4 and formation of the p130/Gsk3beta/beta-catenin complex. AB - Proteins p130 and E2f4, members of the retinoblastoma protein (pRb) family/E2F transcription factor family, are the key elements in regulation of cell cycle and differentiation. The functional role of the p130/E2f4 in mesenchymal stem cells (MSC) is unclear. We demonstrate here that activation of the Wnt/beta-catenin pathway in mouse MSC is associated with accumulation of active forms of the p130, E2f4, and beta-catenin but does not result in inhibition of cell cycle progression. The levels and phosphorylation patterns of p130, E2f4, and beta catenin in MSC do not change during cell cycle progression. This is different from control T98G glyoblastoma cells that accumulated differently phosphorylated forms of the p130 in quiescence, and under active proliferation. In MSC, synchronized at G0/G1 and S cell cycle phases, the p130 and beta-catenin physically interact each other, whereas Gsk3beta was associated and co precipitated with both p130 and beta-catenin. Our results indicate that Wnt/beta catenin and pRb signal pathways interact with each other and form common p130/Gsk3beta/beta-catenin complex during MSC cycle progression. Physiological relevance of such complex may be associated with coupling of the cell cycle and differentiation in MSC, which is related to a wide differentiation potential of these stem cells. PMID- 21631155 TI - A semi-automated micronucleus-centromere assay to assess low-dose radiation exposure in human lymphocytes. AB - PURPOSE: The in vitro micronucleus (MN) assay is a reliable method to assess radiation-induced chromosomal damage in human peripheral blood lymphocytes. It is used to evaluate in vivo radiation over-exposure and to assess in vitro chromosomal radiosensitivity. A limitation of the MN assay is the relatively high and variable spontaneous MN frequency that restricts low-dose estimation to doses of about 0.3 gray (Gy). As radiation-induced MN mainly contain acentric fragments and spontaneous MN originate from lagging chromosomes, both MN types can be distinguished from each other by using fluorescence in situ hybridisation (FISH) with a pan-centromeric probe. The aim of this study was to investigate if the sensitivity, reliability and processing time of the MN assay can be enhanced by combining the automated MN assay with pan-centromere scoring. MATERIALS AND METHODS: Blood samples from 10 healthy donors were irradiated in vitro with low doses of gamma-rays. Dose response curves were determined for fully-automated and semi-automated MN scoring and semi-automated scoring of centromere negative MN (MNCM-). RESULTS: A good correlation was obtained between fully-automated and semi-automated MN scoring (r(2) = 0.9973) and between fully automated MN scoring and semi-automated scoring of MNCM- (r(2) = 0.998). With the Wilcoxon test, a significant p value was obtained between 0 and 0.2 Gy for the fully-automated MN analysis, between 0 and 0.1 Gy for semi-automated MN analysis and between 0 and 0.05 Gy for semi-automated scoring of MNCM-. CONCLUSION: The semi-automated micronucleus-centromere assay combines high-speed MN analysis with a more accurate assessment in the low-dose range which makes it of special interest for large-scale radiation applications. PMID- 21631156 TI - Biomarkers and personalized psychiatry. PMID- 21631157 TI - Pharmacogenetic approaches to cognitive enhancement in schizophrenia. AB - Research in the area of pharmacogenetics in psychiatry is aimed at identifying clinically relevant genetic variations that can predict treatment response. Ultimately, the goal is to individualize treatment in order to optimize outcome in disorders in which incomplete treatment response is common. Positive symptoms in patients with schizophrenia appear to be the most amenable to the currently available agents; however, negative symptoms and cognitive deficits frequently persist even when frank psychosis is well controlled. Given the relationship between these persistent traits and functional disability in schizophrenia, efforts are under way to directly target cognitive impairment and negative symptoms pharmacologically in order to improve quality of life. To date, most pharmacogenetic studies of schizophrenia have been focused on predicting clinical efficacy and side effects. In this review, we discuss the potential use of cognition as a primary outcome measure of interest in future pharmacogenetic trials of schizophrenia. PMID- 21631158 TI - Genes, environment, and individual differences in responding to treatment for depression. AB - A principal weakness of evidence-based psychiatry is that it does not account for the individual variability in therapeutic response among individuals with the same diagnosis. The aim of personalized psychiatry is to remediate this shortcoming and to use predictors to select treatment that is most likely to be beneficial for an individual. This article reviews the evidence that genetic variation, environmental exposures, and gene-environment interactions shape mental illness and influence treatment outcomes, with a primary focus on depression. Several genetic polymorphisms have been identified that influence the outcome of specific treatments, but the strength and generalizability of such influences are not sufficient to justify personalized prescribing. Environmental exposures in early life, such as childhood maltreatment, exert long-lasting influences that are moderated by inherited genetic variation and mediated through stable epigenetic mechanisms such as tissue- and gene-specific DNA methylation. Pharmacological and psychological treatments act on and against the background of genetic disposition, with epigenetic annotation resulting from previous experiences. Research in animal models suggests the possibility that epigenetic interventions may modify the impact of environmental stressors on mental health. Gaps in evidence are identified that need to be bridged before knowledge about cause can inform cure in personalized psychiatry. PMID- 21631159 TI - Glucocorticoids as predictors of treatment response in depression. AB - INTRODUCTION: Glucocorticoids and the stress hormone system have been implicated in the pathophysiology of depression and in the mechanism of action of antidepressant response. Many studies have investigated this system in an effort to predict response to antidepressant treatment. The purpose of this review is to evaluate the evidence for using glucocorticoid-related measures for personalized treatment of depression. METHODS: We conducted a MEDLINE search from 1966 through 2010 and examined English-language studies reporting on the use of endocrine challenge tests and genetic polymorphisms in genes regulating the stress hormone system as predictors of antidepressant response. RESULTS: While measures of glucocorticoid levels using endocrine tests, as well polymorphisms in genes regulating the stress hormone system, show associations with response to antidepressant treatment, these measures will need to be combined with other variables, including clinical information and other biological measures, to realize the goal of highly predictive and clinically relevant biomarkers. DISCUSSION: The glucocorticoid system is potentially of great use in predicting antidepressant response. New combinations of biomarkers including these measures should be tested to develop clinically relevant predictors. PMID- 21631160 TI - Electroencephalography-derived biomarkers of antidepressant response. AB - Recent meta-analyses point to the relatively low efficacy of commonly used antidepressant medications. Selecting the most effective medications for depressed subjects having failed previous treatments is especially difficult. There is a clear need for objective biomarkers that could assist and optimize such treatment selection. We will review here a growing body of evidence suggesting that several electroencephalography (EEG)-based methods may be useful for predicting antidepressant response and eventually for guiding clinical treatment decisions. While most of these methods are based on resting-state EEGs (e.g., alpha- and theta-band EEG abnormalities, the combined Antidepressant Response Index (ATR), cordance, referenced EEG), others include EEG source localization and evoked potentials. The limitations of these technologies and the potential clinical uses will also be outlined. PMID- 21631161 TI - The use of neuroimaging to predict treatment response for neurosurgical interventions for treatment-refractory major depression and obsessive-compulsive disorder. AB - Neuroimaging has contributed profoundly to our understanding of the pathophysiology of psychiatric disorders but has had little impact on treatment. An important goal in neuroscience research is identifying biological markers that predict subsequent response to given treatments. This approach may be especially valuable when considering high-risk and high-cost treatments such as psychiatric neurosurgery. Here, we review neuroimaging findings pertaining to treatment refractory major depression and obsessive-compulsive disorder, and imaging markers that predict response to neurotherapeutic interventions. Thus far, studies conducted with neurotherapeutic interventions have found patterns of predictive brain activity that are similar to those conducted with pharmacological treatments. The predictive neural correlates are concordant with pathophysiological models of major depressive and obsessive-compulsive disorder. These promising findings should motivate additional work establishing the reliability and cost-effectiveness of neuroimaging to predict treatment response across psychiatric diagnoses and interventions. PMID- 21631162 TI - Biomarkers in the psychotherapeutic relationship: the role of physiology, neurobiology, and biological correlates of E.M.P.A.T.H.Y. AB - Emerging biomarker research could powerfully influence the practice of psychotherapy, a standard treatment that is as strongly rooted in brain plasticity as are psychopharmacologic interventions. Psychotherapy is associated with measurable changes in central and peripheral neurophysiology. These markers could be harnessed to aid informed, personalized recommendations for specific psychosocial treatments, to guide a course of treatment, and to predict treatment outcomes, in lieu of relying on costly, trial-and-error approaches. Psychotherapy and empathy research also demonstrate that the patient-doctor relationship has important neurophysiological correlates that can be salient to treatment outcomes, as illustrated in a case example. These correlates include autonomic nervous system arousal manifested by heart rate, respiration rate, muscle tension, and galvanic skin resistance; electroencephalography; and brain-imaging markers. While additional biomarker research is unfolding, there are specific neurobiologically based clinical and subclinical observations, organized by using the E.M.P.A.T.H.Y. mnemonic, that may guide and enhance psychotherapy. Empathic attunement to patients is equally relevant for psychopharmacologic interventions and psychotherapy, and for all patient-doctor relationships. PMID- 21631164 TI - Properties of the target registration error for surface matching in neuronavigation. AB - OBJECTIVE: Surface matching is a relatively new method of spatial registration in neuronavigation. Compared to the traditional point matching method, surface matching does not use fiducial markers that must be fixed to the surface of the head before image scanning, and therefore does not require an image acquisition specifically dedicated for navigation purposes. However, surface matching is not widely used clinically, mainly because there is still insufficient knowledge about its application accuracy. This study aimed to explore the properties of the Target Registration Error (TRE) of surface matching in neuronavigation. MATERIALS AND METHODS: The surface matching process was simulated in the image space of a neuronavigation system so that the TRE could be calculated at any point in that space. For each registration, two point clouds were generated to represent the surface extracted from preoperative images (PC(image)) and the surface obtained intraoperatively by laser scanning (PC(laser)). The properties of the TRE were studied by performing multiple registrations with PC(laser) point clouds at different positions and generated by adding different types of error. RESULTS: For each registration, the TRE had a minimal value at a point in the image space, and the iso-valued surface of the TRE was approximately ellipsoid with smaller TRE on the inner surfaces. The position of the point with minimal TRE and the shape of the iso-valued surface were highly random across different registrations, and the surface registration error between the two point clouds was irrelevant to the TRE at a specific point. The overall TRE tended to increase with the increase in errors in PC(laser), and a larger PC(laser) made it less sensitive to these errors. With the introduction of errors in PC(laser), the points with minimal TRE tended to be concentrated in the anterior and inferior part of the head. CONCLUSION: The results indicate that the alignment between the two surfaces could not provide reliable information about the registration accuracy at an arbitrary target point. However, according to the spatial distribution of the target registration error of a single registration, enough application accuracy could be guaranteed by proper visual verification after registration. In addition, surface matching tends to achieve high accuracy in the inferior and anterior part of the head, and a relatively large scanning area is preferable. PMID- 21631165 TI - Genetic immunization with plasmid DNA mediated by electrotransfer. AB - The concept of DNA immunization was first advanced in the early 1990s, but was not developed because of an initial lack of efficiency. Recent technical advances in plasmid design and gene delivery techniques have allowed renewed interest in the idea. Particularly, a better understanding of genetic immunization has led to construction of optimized plasmids and the use of efficient molecular adjuvants. The field also took great advantage of new delivery techniques such as electrotransfer. This is a simple physical technique consisting of injecting plasmid DNA into a target tissue and applying an electric field, allowing up to a thousandfold more expression of the transgene than naked DNA. DNA immunization mediated by electrotransfer is now effective in a variety of preclinical models against infectious or acquired diseases such as cancer or autoimmune diseases, and is making its way through the clinics in several ongoing phase I human clinical trials. This review will briefly describe genetic immunization mediated by electrotransfer and the main fields of application. PMID- 21631166 TI - In this issue. Editorial. PMID- 21631167 TI - Luteolin inhibits microglial inflammation and improves neuron survival against inflammation. AB - Microglia activation is one of the causative factors for neuroinflammation, which results in brain damage during neurodegenerative disease. Accumulating evidence has shown that the flavonoid luteolin (Lut) possesses potent anti-inflammatory properties; however, its effect on microglia inhibition is currently unknown. Moreover, it is not clear whether Lut also has indirect neuroprotective effects by reducing inflammatory mediators and suppressing microglia activation. In this study, we examined the effects of Lut on lipopolysaccharide (LPS)-induced proinflammatory mediator production and signaling pathways in murine BV2 microglia. In addition, we cocultured microglia and neurons to observe the indirect neuroprotective effects of Lut. Lut inhibited the LPS-stimulated expression of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor alpha (TNF-alpha), and interleukin-1beta (IL-1beta) as well as the production of nitric oxide (NO) and prostaglandin E(2) (PGE(2)). Moreover, Lut blocked LPS-induced nuclear factor kappa B (NF-kappaB) activation. Preincubation of microglia with Lut diminished the neurotoxic effects, owing to the direct anti-inflammatory effects of the compound. Taken together, our findings suggest that Lut may have a potential therapeutic application in the treatment of neuroinflammatory disorders. PMID- 21631168 TI - Influence of intraperitoneal volume on QT dispersion in patients with continuous ambulatory peritoneal dialysis: acute cardiac impact of peritoneal dialysis. AB - AIM: The leading cause of mortality in dialysis patients is cardiovascular complications, including ventricular arrhythmias and sudden cardiac death. QT dispersion (QTd), a simple noninvasive arrhythmogenic marker, is used to assess homogeneity of cardiac repolarization. It was also significantly prolonged in continuous ambulatory peritoneal dialysis (CAPD) patients. The acute cardiac effect of increased abdominal pressure due to infused dialysate during CAPD is not clear yet. In this study we aimed to evaluate corrected QTd (cQTd) and cardiac injury markers such as plasma pro-brain natriuretic peptide (proBNP) and troponin I (TnI) in CAPD patients before and after an infusion of peritoneal dialysate fluid. METHODS: Thirty subjects (16 women, 14 men; mean age, 40.21 +/- 12.34 years) enrolled in our study. QTd, cQTd, maximum QT (QTmax), maximum corrected QT (cQTmax), minimum QT (QTmin), and minimum corrected QT (cQTmin) intervals were measured from standard 12-lead electrocardiography. RESULTS: We found that cQTmax, cQTmin, and cQTd were not changed from baseline measurement after infusion of dialysate in CAPD patients (460 +/- 49 vs. 460 +/- 38, p = 0.9; 410 +/- 36 vs. 410 +/- 41, p = 0.8; 470 +/- 30 vs. 460 +/- 25, p = 0.7, respectively). There were no statistically significant differences between before and after peritoneal dialysate according to the levels of proBNP and TnI (155.64 +/- 76.41 vs. 208.30 +/- 118.46, p = 0.2; 0.008 +/- 0.007 vs. 0.01 +/- 0.011; p = 0.4, respectively). CONCLUSION: In conclusion, we did not find any significant effect of peritoneal dialysate fluid infusion volume on QTd and cardiac injury markers in patients with chronic renal failure receiving CAPD therapy, which is thought to be a safer modality of dialysis. PMID- 21631169 TI - The effect of exercise training on the responsiveness of renal resistance arteries in rats. AB - Blood flow to several tissues changes during an acute bout of exercise. The kidney is one of the organs that are most affected by exercise-induced blood redistribution. The aim of the present study was to investigate possible exercise induced vascular reactivity changes in renal resistance arteries in rats. Renal resistance arteries were isolated from rats that underwent 8 weeks of swimming and sedentary control rats, and the arteries were evaluated using wire myography. Similar dilation responses to acetylcholine, bradykinin, adenosine, isoproterenol, and sodium nitroprusside were observed in both groups. The vasoconstrictive agents vasopressin, endothelin-1, potassium chloride, and thromboxane A(2) also induced similar responses in both groups; however, the trained group had an increased constrictive response to norepinephrine compared to the control rats. The results of our study show that renal resistance arteries of trained rats behave differently than conduit-type renal arteries and exhibit an increased contractile response to sympathetic agonists. This finding provides supporting evidence that renal blood flow markedly decreases during exercise in trained individuals. PMID- 21631170 TI - Renal immaturity mimicking chronic renal failure in an infant born at 22 weeks gestational age. AB - Glomerular function shows a progression directly correlated to gestational age (GA) and postnatal age in preterm infants. In preterms, glomerular filtration rate (GFR) is significantly lower than in term infants, and it matures more slowly in the postnatal period. In infants with very low GA, due to decreased GFR, normalization of serum creatinine values may be prolonged during recovery from acute renal failure (ARF). Herein, we report an extremely premature infant with high creatinine values from third week up to third month of life which normalized thereafter mimicking chronic renal failure. PMID- 21631171 TI - Reliability and validity of Milliken Activities of Daily Living Scale (MAS) in measuring activity limitations of a Turkish population. AB - AIM: The Milliken Activities of Daily Living Scale (MAS) is a self-report scale to address limitation of ability to perform daily tasks in upper extremity injuries. It can address the necessity of each task, which makes MAS useful while planning individual treatment. The objective of this study was to perform cross cultural adaptation of the MAS TURKISH and to evaluate its reliability and validity for Turkish-speaking patients with upper extremity conditions. METHOD: A total of 99 patients were asked to complete the adapted MAS and DASH at baseline and one week after the initial assessment. Also grip strength was evaluated with an interval of one week. RESULTS: The reliability of the adapted version was good, with high internal consistency (Cronbach's alpha = 0.964) and test-retest reliability (interclass correlation coefficient (ICC) = 0.772) for the total score. A statistically significant correlation between MAS and DASH scores and grip strength scores of the injured side was obtained. CONCLUSION: The results of the study have shown that the Turkish version of MAS has excellent test-retest reliability and validity. It is a suitable assessment for evaluating function and giving an overview of activity limitations in many performance areas in a Turkish population. PMID- 21631172 TI - Utility of the Canadian Occupational Performance Measure as an admission and outcome measure in interdisciplinary community-based geriatric rehabilitation. AB - In a community-based geriatric rehabilitation project, the Canadian Occupational Performance Measure (COPM) was used to develop a coordinated, interdisciplinary, and client-centred approach focusing on occupational performance. The purpose of this study was to evaluate the utility of the COPM as an admission and outcome measure in an interdisciplinary geriatric rehabilitation context in Denmark. Eighteen occupational and physiotherapists administered the COPM among elderly citizens. Of 185 citizens referred to the study, 152 were admitted to rehabilitation based on health indices, and 124 completed the COPM after their admission, identifying 404 occupational performance issues in all. Post assessment data were obtained from 95 participants and revealed statistically significant positive change (p < 0.001) in both performance and satisfaction with performance. Furthermore the therapists answered a questionnaire evaluating their experiences, showing that they found development in knowledge and community between the professions to benefit both therapists and citizens, and gained a better insight into their clients' everyday lives through the COPM. In conclusion, the COPM may be useful as an admission and outcome measurement for the rehabilitation of elderly citizens; however, aspects of education and administration must be considered before the instrument can be successfully administered in an interdisciplinary geriatric rehabilitation context. PMID- 21631173 TI - Sharing the agenda: pondering the politics and practices of occupational therapy research. AB - AIMS: Occupational therapists espouse a client-centred philosophy of practice, yet little attention has been given to pondering the politics or client-centred practices of occupational therapy research. The aim of this paper is thus to foster reflection on occupational therapy's commitment to client-centredness in the practice of occupational therapy research. MAJOR FINDINGS: Occupational therapy research is not consistently undertaken in a collaborative manner. Power resides in control of the research agenda and participants' priorities can be supplanted by those of researchers. However, examples from the literature and from the authors' research suggest that study participants may wish to influence the research agenda such that their needs and priorities are addressed. PRACTICE CONCLUSION: Client-centred principles appear to require occupational therapists to undertake collaborative research and to ensure that research agendas are informed by clients' priorities. Commitment to client-centred principles demands concerted efforts to identify and address potential barriers to meaningful client participation in the occupation of research. However, it is argued that if researchers and disabled people collaborate, and pool their knowledge and expertise, they may achieve research that is more philosophically compatible with espoused professional values; and that collaborative research may also inform more relevant and useful client-centred clinical practices. PMID- 21631174 TI - User involvement in mental health rehabilitation: a struggle for self determination and recognition. AB - AIM: The study aimed to explore service user involvement in supported housing schemes as experienced by adults with mental illness in interplay with professionals during rehabilitation. METHOD: The study was designed as a field study in two supported housing schemes, using an ethnographic approach to data collection, including participant observation, informal conversations, and individual and group interview. Twelve participants aged 22 to 45 were included. The data analysis was informed by Ricoeur's theory of textual interpretation, and followed his three levels of interpretation: naive reading, structural analysis, and critical interpretation. FINDINGS: The study showed that user involvement in rehabilitation was experienced by the users as a struggle for self-determination and recognition. Being able to decide and to have influence on daily activities and everyday life, together with having goals to reach during rehabilitation, were viewed as valuable. Situations without influence or involving coercion were reported especially during periods of illness or hospitalization. CONCLUSIONS: The results of this study contribute to knowledge about user involvement from service users' perspective, and highlight the difficulties experienced in achieving self-determination and recognition during rehabilitation. PMID- 21631175 TI - Occupational therapists' practice patterns for clients with cognitive impairment following acquired brain injury: development of a questionnaire. AB - Clients with cognitive impairment following acquired brain injury (CIABI) are a common group to receive occupational therapy services. Research has shown that occupational therapy has a positive effect on occupational performance for these clients, but the exact nature of the interventions is not clearly described and needs to be better understood and defined. The aim of this study was to develop and evaluate an empirically derived questionnaire for the purpose of surveying occupational therapists' (OTs') practice patterns in relation to CIABI. The questionnaire was developed from the results of a former qualitative study. It was evaluated for content validity by a group of six OT researchers with experience in CIABI, using the content validity index (CVI). Reliability was evaluated by a test-retest design with a group of 51 OTs. Data were analysed by non-parametric statistical methods. Initially the questionnaire consisted of 90 items dealing with OT practice and nine demographic questions. After the reliability and content validity process the OT practice items were reduced to 44. The revised questionnaire will be used to survey and explicitly describe occupational therapy practice for clients with CIABI. PMID- 21631176 TI - Height loss caused by bent posture: a risk factor for stroke from ENT clinic - is it time to reconsider the physical examination? AB - CONCLUSION: When excessive height loss occurs in the elderly, which may be indicated by a cervical-bent posture, examination of the head and neck should be performed to detect common carotid artery (CCA) and internal carotid artery (ICA) aberration. In such cases brain magnetic resonance imaging (MRI) examination should be conducted to determine whether infarction is present. OBJECTIVES: To examine the relationships among bent posture, height loss, aberration of carotid arteries, and ischemic stroke risk with a case-control study. METHODS: Controls (n = 163) were selected from among patients who had undergone MRI of the brain because of otolaryngologic symptoms. Case patients (n = 72) were selected from among those whose primary diagnosis was cerebral infarction in the area served by the ICA. Both groups were age-matched between 65 and 84 years old. The neck and pharyngeal cavity in each of the 235 patients were examined to determine whether the carotid arteries exhibited aberration. Patients' current height and greatest lifetime height were recorded, along with presence or absence of bent posture and other stroke risk factors. RESULTS: Height loss alone could predict stroke risk in 79.1% of patients: 90.2% based on carotid artery aberration and 91.4% when all risk factors (aberration of carotid artery, height loss, bent posture) were analyzed. PMID- 21631177 TI - Head and neck cancer in young adults and nonsmokers: study of cancer susceptibility by genome-wide high-density SNP microarray mapping. AB - CONCLUSION: Our results raise the question as to whether specific patterns of 'germline loss of heterozygosity (LOH)' could contribute to the genetic susceptibility for head and neck squamous cell carcinoma (HNSCC). OBJECTIVES: HNSCC usually occurs in older individuals with a history of smoking. However, about 5% of HNSCC patients have never used tobacco or develop this disease at an exceptionally young age. Therefore, genetic susceptibility must contribute significantly to HNSCC risk. The objective was to introduce a novel approach that might help to unveil candidate genes contributing to cancer predisposition and to identify individuals at risk for HNSCC, and to present our observations with this method in a specific group of patients. METHODS: High-resolution SNP (single nucleotide polymorphism) microarray mapping for homozygous stretches in germline DNA was performed in 12 patients who appeared particularly susceptible to develop HNSCC, because they were exceptionally young or never users of tobacco. RESULTS: We could identify strings of consecutive homozygous SNPs that were much longer than would be expected to appear by chance alone, indicating regions of DNA deletions that we named germline LOH. PMID- 21631178 TI - Age-related hearing loss and expression of antioxidant enzymes in BDF1 mice. AB - CONCLUSION: Our data suggest that the changes in expression of antioxidant enzymes may cause age-related hearing loss (AHL). OBJECTIVES: AHL is an aging process of the inner ear, and oxidant stressors are considered to be one of the leading causes. We investigated the hearing level and expression profile of antioxidant enzymes in aged mice. METHODS: Mice aged 3, 6, and 11 months were used. Hearing levels of the mice were examined using the auditory brainstem response (ABR). After measuring the ABR threshold, cochleae were dissected. RNA was isolated from the cochleae, and cDNA was synthesized using the retro transcription enzyme. Expression of the antioxidant enzymes was measured by quantitative real-time PCR. RESULTS: The ABR thresholds of the BDF1 mice were elevated by 6 months of age. The expression of superoxide dismutase 1 (SOD1) and heme oxygenase 1 (HO1) at 11 months of age significantly decreased compared with that of those at 6 months of age. In contrast, a decrease in the expression level was not observed regarding NAD(P)H-quinone oxidoreductase 1 (NQO1). PMID- 21631179 TI - Four systems involved with congenital abnormalities: a new type of syndromic hearing loss - ADOC Wang's syndrome? AB - Syndromic hearing impairment encompasses hundreds of phenotypes. We identified a young female patient affected by the unique combination of dysplasia of the auricular system, patent ductus arteriosus (PDA), choroideremia, and enamel hypoplasia. The patient was treated with PDA ligature and left exploratory tympanotomy. Impairment in all four systems suggests a correlation with the neural crest. It is presumed that all of the features result from the same origin, probably through autosomal recessive inheritance or a novel mutation during the embryonic period. When audio-dento-oculo-cardio systems are involved, we suggest that this new syndrome can be named 'ADOC Wang's syndrome', summarizing the disorders of the four systems and indicative of the founding person (Dr Wang, the first and corresponding author of the paper). PMID- 21631180 TI - Radix Astragali injection enhances recovery from acute acoustic trauma. AB - CONCLUSION: The average recovery of hearing and cessation of tinnitus was significantly better after treatment with Radix Astragali (RA) than after non treatment with RA. RA can be valuable adjuvant therapy for patients with acute acoustic trauma (AAT). OBJECTIVES: AAT is one of the early indications for the use of RA. The reasons for administering RA to patients with AAT are based on experimental studies showing that noise exposure results in the formation of reactive oxygen species (ROS), which trigger metabolic damage to the organ of Corti. RA is a natural antioxidant. The aim of this study was to investigate the efficacy of RA in patients with AAT. METHODS: We compared the recovery from hearing impairment and tinnitus in 40 ears treated with RA with 40 ears treated with non-RA. RA was given intravenously daily for 10 days. There were no significant differences in clinical or audiological data between RA and non-RA groups. RESULTS: The average recovery of hearing at both high and speech frequencies was significantly better and tinnitus persisted less commonly in the RA group than in the non-RA group. Normal hearing at the end of the follow-up period was regained in 27 ears in the RA group and in 21 ears in the non-RA group (p < 0.01). PMID- 21631181 TI - Tumor necrosis factor-alpha promoter gene polymorphisms are not associated with hepatitis C virus infection in Chinese hemodialysis patients. AB - BACKGROUND AND AIMS: The aim of this study was to investigate the possible influence of tumor necrosis factor-alpha (TNF-alpha) gene promoter polymorphisms on the hepatitis C virus (HCV) infection in Chinese hemodialysis (HD) patients. METHODS: A total of 884 HD patients from 14 HD centers in south China were investigated. The TNF-alpha gene promoter polymorphisms at positions 238 and 308 of the patients were detected by the polymerase chain reaction (PCR)-restriction fragment length polymorphism method. RESULTS: Among the 884 patients, 176 (19.9%) were anti-HCV (+), and 142 (80.7%) of the anti-HCV (+) patients became chronically infected. The anti-HCV (+) patients showed longer duration of HD, higher rate of blood transfusion, kidney transplantation, and dialyzer reuse, compared with the anti-HCV (-) patients. However, the distributions of TNF-alpha 238 and -308 alleles and genotypes had no significant differences between the anti-HCV (+) and the anti-HCV (-) patients (p > 0.05). And the frequencies of the above alleles and genotypes were also approximately equally distributed in the persistent infection and in the viral clearance HD patients (p > 0.05). CONCLUSIONS: This study did not suggest that the TNF-alpha -238 and -308 polymorphisms had influence on the infection of HCV in Chinese HD patients. PMID- 21631182 TI - Toxicodynamic evaluation of a cisplatin-chondroitin sulfate complex using a perfused kidney and human proximal tubular cells. AB - Cisplatin (CDDP) is an anticancer drug. The clinical limitations associated with CDDP have stimulated the development of macromolecular drug-carrier systems, in attempts to decrease its toxicity. A complex (CDDP-CSA-23) between CDDP and chondroitin sulfate (CSA), a natural polysaccharide with a mean molecular weight of 23 kDa, proved to have the same anticancer activity as CDDP. A toxicodynamic study was performed on perfused kidneys to determine the effect of CDDP-CSA-23 on renal functions and the extent of platinum accumulation. The results showed that CDDP-CSA-23 attenuates the reduction in urine flow and creatinine clearance induced by CDDP. Moreover, significantly lower amounts of platinum were excreted into the urine in the case of CDDP-CSA-23, compared with CDDP alone. Meanwhile, CDDP-CSA-23 effectively retarded the rapid perfusion of platinum into kidney tissues, as occurs when CDDP is being perfused alone. The cytoprotective effects of CDDP-CSA on human proximal tubular (HK-2) cells were examined by measuring the growth of HK-2 cells in the presence of CDDP or CDDP-CSA-23. Interestingly, CDDP CSA-23 was found to have a significantly reduced cytotoxicity, compared to CDDP. These results suggest that CDDP-CSA-23 greatly decreased the negative effects of CDDP on glomerular filtration and tubular transport in kidneys at early stages of its administration. PMID- 21631183 TI - Children's longing for everydayness: life following traumatic brain injury in the USA. AB - PRIMARY OBJECTIVE: Little is known about life after traumatic brain injury (TBI) from the child's perspective. RESEARCH DESIGN: This descriptive phenomenological investigation explored themes of children's experiences following moderate-to severe TBI. METHODS AND PROCEDURES: INCLUSION CRITERIA: (1) 6-18 years of age at injury; (2) moderate-to-severe TBI; (3) <=3 years since injury; and (4) English speaking and could participate in an interview. Children participated (n = 39) in two interviews at least 1 year apart. A preliminary model was developed and shared for participants' input. MAIN OUTCOMES AND RESULTS: Six themes emerged: (1) it is like waking up in a bad dream; (2) I thought going home would get me back to my old life, but it did not; (3) everything is such hard work; (4) you feel like you will never be like the person you were before; (5) it is not all bad; and (6) some people get it, but many people do not. CONCLUSIONS: Social support was important to how children adjusted to changes or losses. Most children did adjust to functional changes by second interviews. Children had a more difficult time adjusting to how others defined them and limited their possibilities for a meaningful life. PMID- 21631184 TI - Severity and outcome of traumatic brain injuries (TBI) with different causes of injury. AB - INTRODUCTION: Most epidemiological studies and reports have reached a consensus on the leading causes of traumatic brain injury (TBI). Despite the fact that the area of TBI is relatively well studied, reports on differences in severity and outcome of TBI with different causes are lacking. GOAL: This paper analyses the differences in severity and in short- and long-term outcome of TBIs with different causes. METHOD: This study analysed data on 1109 patients with severe TBI by dividing them into three injury-cause groups: 'traffic-related', 'falls' and 'other causes'. Severity and outcome was evaluated using chosen direct and indirect indicators. RESULTS: The most severe trauma occurred in the traffic related group followed by falls and injuries with other causes. On the other hand, patients with traffic-related TBI had the best outcome. Age improves the outcome in the traffic-related group significantly. However, in the multivariate analysis after adjusting for age (and other important predictors including level of care) the odds for favourable long-term outcome stayed significantly higher in the traffic-related group. CONCLUSIONS: It is concluded that the causes of TBI should be considered by both clinicians and public health professionals as a lead in prognosis of outcome and policy planning. PMID- 21631186 TI - Balint's syndrome and post-acute brain injury rehabilitation: a case report. AB - BACKGROUND: Balint's syndrome includes the clinical symptom triad of simultagnosia, ocular apraxia and optic ataxia. These symptoms, in combination, are rare and can be quite debilitating as they impact visuospatial skills, visual scanning and attentional mechanisms. CASE STUDY: The literature addressing rehabilitation of individuals with Balint's syndrome is sparse. The current case report describes the outcome of a 58-year old male who presented with Balint's syndrome secondary to severe traumatic brain injury and following completion of a comprehensive post-acute brain injury rehabilitation programme. The patient was 4 months post-injury onset upon admission and received 6 months of rehabilitation services as an inpatient. The patient's comprehensive rehabilitation programme involved a 3-pronged approach including the implementation of (a) compensatory strategies, (b) remediation exercises and (c) transfer of learned skills in multiple environments and situations with implementation of psychoeducation and psychotherapy. Comprehensive neuropsychological and occupational therapy evaluations were performed at admission and at discharge in order to monitor cognitive, affective, neurological and functional change over time. CONCLUSIONS: Neuropsychological test improvements were noted on tasks that assess visuospatial functioning, although most gains were noted for functional and physical abilities. PMID- 21631187 TI - Effects of exposure to static magnetic field on motor skills and iron levels in plasma and brain of rats. AB - PRIMARY OBJECTIVE: The present work investigated the behavioural and biochemistry effects of moderate exposure to a static magnetic field (SMF) in rats. SMF effects were evaluated in sham- and SMF-exposed rats. METHODS: Adult Wistar rats were exposed for 1 hour per day for 5 consecutive days to 128 millitesla (mT) SMF. Then, their motor skills were tested using a Stationary beam and Suspended string test. Iron level in plasma and brain (i.e. frontal cortex, basal ganglia, hippocampus and cerebellum) was measured. RESULTS: No significant change was observed between sham and SMF-exposed rats in the Stationary beam and Suspended string test. However, the same treatment induced an increase in plasma transferrin content (+25.4%) and decreased the iron level in plasma (-16.2%). The SMF treatment failed to alter the iron concentration in the brain. CONCLUSION: The findings indicate that SMF exposure induced iron deficiency in plasma but did not induce motor-skills deficit. PMID- 21631188 TI - Resilience and the mediating effects of executive dysfunction after childhood brain injury: a comparison between children aged 9-15 years with brain injury and non-injured controls. AB - PRIMARY OBJECTIVE: Acquired brain injury (ABI) during childhood can be associated with enduring difficulties related to impairments to executive functioning (EF). EF impairments may detrimentally affect outcome by restricting an individual's ability to access 'resiliency' resources after ABI. RESEARCH DESIGN: The purpose of this study was to explore whether there is deterioration in children's resilience compared with peers after ABI and whether EF is influential in mediating relationships between resilience and behaviour. METHODS AND PROCEDURES: Measures of resilience, depression and anxiety were administered with 21 children with ABI and 70 matched healthy children aged 9-15 years. Parents completed measures of behaviour and EF. MAIN OUTCOMES AND RESULTS: Children with ABI were identified as less resilient and more depressed and anxious than controls. Resiliency measures were correlated with depression and anxiety in both groups. Relationships between resiliency and socio-emotional behaviour were mediated by EF. CONCLUSIONS: Assessment of resilience after ABI may be useful in supporting or defining the delivery of more individualized rehabilitation programmes according to the resources and vulnerabilities a young person has. However, an accurate understanding of the role of EF in the relationship between resilience and behavioural outcome after ABI is essential. PMID- 21631189 TI - Participation in leisure activities during brain injury rehabilitation. AB - PRIMARY OBJECTIVE: To describe and compare pre- and post-injury leisure activities of individuals receiving brain injury rehabilitation and explore levels of leisure participation and satisfaction. RESEARCH DESIGN: Cross sectional descriptive study incorporating a survey of current and past leisure activities. METHODS AND PROCEDURES: Questionnaires were completed by 40 individuals with an acquired brain injury receiving inpatient or outpatient rehabilitation. MEASURES: Shortened Version of the Nottingham Leisure Questionnaire and Changes in Leisure Questionnaire (developed for this study). RESULTS: Leisure participation declined following injury, particularly in social leisure activities. Pre-injury activities with high rates of discontinued or decreased participation were driving, going to pubs and parties, do-it-yourself activities and attending sports events. Inpatient participants generally attributed decreased participation to the hospital environment, whereas outpatient participants reported this predominantly as a result of disability. Post-injury levels of perceived leisure satisfaction were significantly lower for the inpatient group compared to pre-injury, but not for the outpatient group. Uptake of some new leisure activities was reported post-injury, however not at the rate to which participation declined. CONCLUSIONS: Leisure participation decreases during brain injury rehabilitation compared to pre-injury levels. Re engagement in relevant, age-appropriate leisure activities needs to be addressed during rehabilitation to improve participation in this domain. PMID- 21631191 TI - Intracranial suppuration complicating sinusitis among children: an epidemiological and clinical study. AB - OBJECT: Intracranial abscess is a very infrequent complication of sinusitis among children. Case series, mostly small and focusing on surgical treatment, have appeared in the literature, but there has been no study on the epidemiology of this condition, nor has there been any large-scale study on the intensity of therapy or outcomes. METHODS: Data were drawn from the Kids' Inpatient Databases for 1997, 2000, 2003, and 2006 and the National Inpatient Sample for 2001. Cases were defined by the conjunction between the diagnostic codes for intracranial abscess and those for acute or chronic sinusitis or mastoiditis. RESULTS: Eight hundred forty-seven hospital admissions were captured. Over the 10 years of the study, admission rates ranged between 2.74 and 4.38 per million children per year. Boys were affected much more commonly than girls. Black children were affected out of proportion to their presence in the population. The overall incidence seemed to peak in early adolescence. Sinogenic cases had a marked seasonal pattern peaking in winter, but no seasonal variation was seen for otogenic cases. Asthma comorbidity was more prevalent among sinogenic cases. The mortality rate was 2%, and death occurred only among sinogenic cases. Moreover, sinogenic cases tended to require more intensive therapy, as measured by the number of procedures, and there was a trend toward less favorable discharge dispositions. Older patients and black patients were less likely to be discharged directly to home. At least a quarter of the cases were managed without neurosurgical intervention. White patients were treated without neurosurgery more often than others. CONCLUSIONS: Analysis of administrative data sets has yielded a descriptive picture of intracranial abscess complicating sinusitis among children, but the very low incidence of this condition impedes prospective clinical research directed at practical management issues. PMID- 21631192 TI - Harvey Cushing and pediatric brain tumors at Johns Hopkins: the early stages of development. AB - OBJECT: Harvey Cushing, credited with pioneering the field of neurosurgery as a distinct surgical subspecialty in the US, was at the forefront of neurooncology, publishing seminal papers on the diagnosis and treatment of pediatric brain tumors during the latter part of his career. However, his contributions to the surgical treatment of these lesions during the early stages of his tenure at the Johns Hopkins Hospital, from 1896 to 1912, remain largely unknown. METHODS: After obtaining institutional review board approval, and through the courtesy of the Alan Mason Chesney Archives, the authors reviewed the Johns Hopkins Hospital surgical files from the years 1896 to 1912. Patients younger than 18 years of age, presenting with symptoms suspicious for an intracranial tumor, and undergoing surgical intervention by Cushing were selected for further analysis. RESULTS: Of the 40 pediatric patients undergoing surgery for suspected intracranial neoplasms, 26 were male. The mean age among the entire sample was 10.1 years. Cushing used three main operative approaches in the surgical treatment of pediatric intracranial neoplasms: infratentorial/suboccipital, subtemporal, and hemisphere flap. Twenty-three patients had negative findings following both the primary and subsequent surgical interventions conducted by Cushing. Postoperative conditions following the primary surgical intervention were improved in 24 patients. Twelve patients (30%) died during their inpatient stay for the primary intervention. The mean time to the last follow-up was 24.9 months; the mean time to death was 10.0 months. CONCLUSIONS: Cushing strove to maximize exposure while minimizing blood loss in an attempt to increase his ability to successfully treat pediatric brain tumors. His early contributions to the field of pediatric neurooncology demonstrate his commitment to advancing the field of neurosurgery. PMID- 21631193 TI - Surgical management of tumors producing the thalamopeduncular syndrome of childhood. AB - OBJECT: Thalamopeduncular tumors arise at the junction of the inferior thalamus and cerebral peduncle and present with a common clinical syndrome of progressive spastic hemiparesis. Pathologically, these lesions are usually juvenile pilocytic astrocytomas and are best treated with resection with the intent to cure. The goals of this study are to define a common clinical syndrome produced by thalamopeduncular tumors and to discuss imaging characteristics as well as surgical adjuncts, intraoperative nuances, and postoperative complications relating to the resection of these neoplasms. METHODS: The authors present a retrospective review of their experience with 10 children presenting between 3 and 15 years of age with a thalamopeduncular syndrome. Formal preoperative MR imaging was obtained in all patients, and diffusion tensor (DT) imaging was performed in 9 patients. Postoperative MR imaging was obtained to evaluate the extent of tumor resection. A prospective analysis of clinical outcomes was then conducted by the senior author. RESULTS: Pilocytic astrocytoma was the pathological diagnosis in 9 cases, and the other was fibrillary astrocytoma. Seven of 9 pilocytic astrocytomas were completely resected. Radical surgery was avoided in 1 child after DT imaging revealed that the corticospinal tract (CST) coursed through the center of the tumor, consistent with the infiltrative nature of fibrillary astrocytoma as identified by stereotactic biopsy. In 8 patients, tractography served as an important adjunct for designing a surgical approach that spared the CST. In 6 cases the CSTs were pushed anterolaterally, making a transsylvian approach a poor choice, as was evidenced by the first patient in the series, who underwent operation prior to the advent of tractography, and who awoke with a dense contralateral hemiparesis. Thus, subsequent patients with this deviation pattern underwent a transcortical approach via the middle temporal gyrus. One patient exhibited medial deviation of the tracts and another had lateral deviation, facilitating a transtemporal and a transfrontal approach, respectively. CONCLUSIONS: The thalamopeduncular syndrome of progressive spastic hemiparesis presenting in children with or without symptoms of headache should alert the examiner to the possibility of a tumoral involvement of CSTs. Preoperative tractography is a useful adjunct to surgical planning in tumors that displace motor pathways. Gross-total resection of pilocytic astrocytomas usually results in cure, and therefore should be entertained when developing a treatment strategy for thalamopeduncular tumors of childhood. PMID- 21631194 TI - Interhemispheric endoscopic resection of large intraventricular and thalamic tumors. AB - A transfrontal route is the traditional endoscopic approach to intraventricular tumors. Small lesions can be biopsied using the parallel port channel of the endoscope. For larger tumors a ventriculoport can be used for resection. This technique nevertheless requires traversing the brain tissue, is difficult in the setting of small ventricles, and allows only limited mobility. The authors describe the endoscopic resection of large intraventricular tumors via an interhemispheric route using rigid suction with a mounted endoscope, and thus circumventing some of the problems with the traditional approach. PMID- 21631195 TI - Endoscopically assisted intratumoral embolization of juvenile nasopharyngeal angiofibroma using Onyx. AB - Juvenile nasopharyngeal angiofibromas are vascular tumors that may make resection difficult and potentially dangerous. Preoperative embolization is frequently used to decrease surgical morbidity and blood loss. Embolization has typically been performed via a transarterial route using a variety of embolic materials. The authors present a case in which endoscopic assistance was used for direct transnasal tumor puncture and intratumoral embolization using the liquid embolic agent Onyx. In this case there was excellent infiltration of the parenchymal vasculature with complete angiographic obliteration. There were no complications related to the embolization. The tumor was resected with minimal blood loss. To the authors' knowledge, there have been no previous reports of this novel direct intratumoral embolization technique using endoscopic guidance. PMID- 21631196 TI - Intracranial yolk sac tumor in a patient with Down syndrome. AB - The authors report a rare case of intracranial yolk sac tumor in a 13-year-old boy with Down syndrome who presented with left hemiparesis. Admission MR imaging revealed a tumor in the right basal ganglia. Serum alpha-fetoprotein was markedly elevated. Yolk sac tumor was diagnosed radiologically and serologically. The standard therapy for intracranial yolk sac tumor is platinum-based chemotherapy with concomitant radiotherapy. However, the authors used reduced-dose chemotherapy and asynchronized radiotherapy because of the well-known low tolerance of patients with Down syndrome to chemotherapy. This treatment was successful with no complications. Blood cancers are frequently associated with Down syndrome, whereas solid tumors occur less frequently in these patients, and the risk of chemoradiotherapy is unclear. The results indicate that dose reduction therapy can be effective for treatment of a brain tumor in a patient with Down syndrome. PMID- 21631197 TI - Perioperative seizure incidence and risk factors in 223 pediatric brain tumor patients without prior seizures. AB - OBJECT: The incidence of, and risk factors for, perioperative seizures and the need for perioperative antiepileptic drugs (AEDs) in previously seizure-free children with brain tumors remains unclear. The authors have undertaken a review of previously seizure-free pediatric patients with brain tumors undergoing resection to identify the incidence of seizures in the perioperative period, and to characterize risk factors for perioperative seizures in this population. METHODS: A retrospective review was conducted of all patients between 0 and 19 years of age without prior seizures who underwent intracranial tumor resection at the authors' institution between January 2005 and December 2009. RESULTS: Of the 223 patients undergoing 229 operations, 7.4% experienced at least 1 clinical seizure during the surgical admission. Over half of all tumors were supratentorial. Only 4.4% of patients received prophylactic AEDs. Independent factors associated with perioperative seizures included supratentorial tumor, age < 2 years, and hyponatremia due to syndrome of inappropriate antidiuretic hormone or cerebral salt wasting. Tumor type, lobe affected, operative blood loss, and length of surgery were not independently associated with seizure incidence. CONCLUSIONS: Perioperative seizures in previously seizure-free children undergoing resection of brain tumors are associated with supratentorial tumor location, age < 2 years, and postoperative hyponatremia. Perioperative seizures are not associated with tumor pathology, tumor size, or frontotemporal location. Due to the low incidence of seizures in this series in patients more than 2 years old with normal serum sodium, the authors recommend that pediatric patients with brain tumors not routinely receive perioperative prophylactic AEDs. However, the role for prophylaxis in patients younger than 2 years of age deserves further study. PMID- 21631198 TI - Minimally invasive lumbar microdiscectomy in pediatric patients: a series of 6 patients. AB - OBJECT: Lumbar disc herniation is rare in the pediatric age group, but may still cause a significant amount of pain and disability. Whereas minimally invasive surgery (MIS) for lumbar disc herniation is routinely performed in adults, it has not yet been described in the pediatric population. The purpose of this study was to describe the surgical results of pediatric MIS-treated lumbar disc disease. METHODS: The authors retrospectively reviewed a series of 6 consecutive cases of lumbar microdiscectomy performed using MIS techniques between April 2008 and July 2010. Presenting symptoms, physical examination findings, and preoperative MR imaging results were obtained from medical records. Perioperative results, including blood loss, length of hospital stay, and complications were assessed. Findings at latest follow-up evaluation were also recorded. RESULTS: This report represents the first surgical series regarding pediatric lumbar microdiscectomies performed using MIS. The mean patient age was 16 years (range 14-17 years); there were 4 girls and 2 boys. Preoperative signs and symptoms of radiculopathic pain were notable in 100% of patients, and myotomal weakness was noted in 33% of patients. The first line of treatment in all patients was a period of conservative management lasting an average of 11.5 months (range 6-12 months). The mean intraoperative blood loss was 10.8 ml, and the mean postoperative length of stay was 1.3 days. There were no complications in this small series. CONCLUSIONS: The treatment of pediatric lumbar disc herniation by using MIS techniques can be safe and efficacious. However, further study with a larger number of patients and longer follow-up is needed to compare outcomes between MIS and open microdiscectomies. PMID- 21631199 TI - The management of sagittal synostosis using endoscopic suturectomy and postoperative helmet therapy. AB - OBJECT: Suturectomy as a treatment for craniosynostosis was largely replaced in the late twentieth century by more extensive, but predictable, cranial remodeling procedures. Recent technical innovations, such as using the endoscope combined with postoperative orthotic reshaping, have led to a resurgence of interest in suturectomy as a safer, less invasive method. METHODS: A retrospective chart review was performed for all cases of sagittal synostosis treated with endoscopic sagittal suture strip craniectomy and helmet therapy between 2004 and 2008. Data collected included gestational age, genetic evaluations and syndromic status, age at operation, duration of procedure, need for blood transfusions, length of hospital stay, preoperative and postoperative head circumference percentile and cranial index, duration of helmet use, length of follow-up, complications, and revisions. RESULTS: Fifty-six patients with isolated sagittal synostosis were treated using endoscopic suturectomy and completed helmet therapy. Mean age at time of procedure was 3.24 months. Mean operative duration was 45.32 minutes. Mean hospital stay was 1.39 days. There were 2 transfusions and no deaths. The mean length of follow-up was 2.34 years. Helmet therapy was instituted for a mean of 7.47 months. Head circumference percentile increased from 61.42% to 89.27% over 2 years of follow-up. Cranial index increased from a preoperative mean of 0.69 to 0.76 over 2 years of follow-up. Reoperations for synostosis included 1 sagittal suture refusion and 2 cases in which other sutures fused. CONCLUSIONS: Sagittal synostosis can be safely treated with endoscopic suturectomy and helmet therapy. Improvements in cranial volume and shape are comparable to open procedures and are enduring. PMID- 21631200 TI - Effect of molding helmet on head shape in nonsurgically treated sagittal craniosynostosis. AB - OBJECT: Sagittal craniosynostosis is traditionally considered to be a surgical condition. Poor results of simple suturectomy follow from early reclosure of the suture. A wider craniectomy or use of interposing materials has not improved the outcome. However, endoscopic suturectomy supplemented with postoperative use of a molding helmet has shown good results. Because suturectomy reunites within 8-12 weeks of surgery, the authors questioned if the improved outcome was primarily related to use of the helmet. METHODS: In 4 patients whose families opted for calvarial reconstruction when the infant was 4-6 months old, instead of endoscopic suturectomy, a molding helmet was used to minimize compensatory changes in the interim. Patients underwent 3D CT scanning to confirm craniosynostosis. Follow-up visits were made at intervals of 4 weeks for adjustment of the helmet, head circumference measurements, clinical photographs, and cranial index measurement. RESULTS: There was significant improvement in the head shape within 6 weeks of use of the molding helmet. The cranial index score improved from a mean (+/- SD) of 67% +/- 3% to 75% +/- 2%. CONCLUSIONS: These cases demonstrate that molding helmets improve head shape even without a suturectomy in patients with sagittal craniosynostosis, challenging the traditional view. PMID- 21631201 TI - Tumor bleeding from a de novo aneurysm associated with optic glioma. AB - Aneurysms in children are rare and potentially devastating lesions. The authors report the case of a 16-year-old girl with a complicated medical history related to a chiasmal glioma diagnosed at 18 months of age. She had previously received multiple modalities of radiation treatment, including external beam, proton therapy, and Gamma Knife. She presented with hemorrhage centered in the tumor and extending into the ventricular space. There was no subarachnoid blood. Magnetic resonance imaging demonstrated the hemorrhage and tumor anatomy. Magnetic resonance angiography revealed an aneurysm at the internal carotid artery bifurcation, but the lesion was more clearly delineated on CT angiography. A comparison MR imaging study obtained 6 months earlier, even in retrospect, did not show evidence of an aneurysm. This case illustrates the salient point that the clinician must search for vascular lesions in the patient with spontaneous "tumor bleeding," especially if that patient has risk factors for aneurysm formation. The authors also suggest that a CT angiogram is better at radiographically demonstrating an intratumoral aneurysm than an MR angiogram in this scenario. PMID- 21631202 TI - Balloon-assisted Onyx embolization of cerebral single-channel pial arteriovenous fistulas. AB - OBJECT: Pial arteriovenous fistulas (AVFs) of the brain are rare vascular malformations associated with significant risks of hemorrhage and neurological deficit. Depending on their location and high-flow dynamics, these lesions can present treatment challenges for both endovascular and open cerebrovascular surgeons. The authors describe a novel endovascular treatment strategy that was used successfully to treat 2 pediatric patients with a pial AVF, and they discuss the technical nuances specific to their treatment strategy. METHODS: A single channel high-flow pial AVF was diagnosed in 2 male patients (6 and 17 years of age). Both patients were treated with endovascular flow arrest using a highly conformable balloon followed by Onyx infusion for definitive closure of the fistula. RESULTS: Neither patient suffered a complication as a result of the procedure. At the 6-month follow-up in both cases, the simple discontinuation of blood flow had resulted in durable obliteration of the fistula and stable or improved neurological function. CONCLUSIONS: Onyx can be delivered successfully into high-flow lesions after flow arrest to allow a minimally invasive and durable treatment for pial AVFs. PMID- 21631203 TI - Complications of endoscopic third ventriculostomy. AB - OBJECT: Endoscopic third ventriculostomy (ETV) is an established treatment for hydrocephalus. Most studies focus on success rate, and complications are insufficiently documented. The aim of this study was to perform a systematic review of ETV complications. METHODS: A Medline search discovered 34 series of ETV with detailed complications reports (17 series involving exclusively pediatric patient populations, 6 series involving exclusively adults, and 11 series involving mixed adult and pediatric populations). RESULTS: The analysis included 2985 ETVs performed in 2884 patients. The cause of hydrocephalus was aqueductal stenosis in 29.3% of patients, tumor in 37.6%, meningomyelocele in 7.6%, cysts in 2.6%, cerebellar infarct in 0.9%, Dandy-Walker malformation in 0.6%, and Chiari malformation Type I in 0.4%; 7.4% of the patients had posthemorrhagic hydrocephalus, 1.8% had postinfectious hydrocephalus, and 1.2% had normal pressure hydrocephalus. Hydrocephalus was due to other causes in 1.3% of cases and the cause was not reported in 9.8%. The overall complication rate was 8.5%. The rate of permanent morbidity was 2.38%; the rate of permanent neurological complications was 1.44% (hemiparesis, gaze palsy, memory disorders, altered consciousness), and the rate of permanent hormonal morbidity was 0.94% (diabetes insipidus, weight gain, precocious puberty). The rate of intraoperative hemorrhage rate was 3.7%; the rate of severe intraoperative hemorrhage was 0.6% (including a 0.21% rate of basilar rupture). The rate for intraoperative neural injury (thalamic, forniceal, hypothalamic, and midbrain injuries) was 0.24%. Central nervous system infections occurred in 1.81% of cases, CSF leak in 1.61%, and postoperative intracranial hematomas in 0.81% of cases. The early postoperative mortality rate was 0.21% (6 patients died; 2 of sepsis and 4 of hemorrhage). Another 2 children suffered delayed "sudden death" (one after 25 months and the other after 60 months), caused by acute hydrocephalus due to stoma occlusion. There were no significant differences between series involving pediatric or adult patient populations or series with fewer than 100 or more than 100 patients. All reported deaths were in series involving more than 100 patients. CONCLUSIONS: Endoscopic third ventriculostomy can be regarded as a low complication procedure, with an overall complication rate of 8.5%, permanent morbidity rate of 2.4%, mortality rate of 0.21%, and delayed "sudden death" rate of 0.07%. PMID- 21631204 TI - Bilateral subthalamic nucleus deep brain stimulation for dopa-responsive dystonia in a 6-year-old child. AB - Tyrosine hydroxylase (TH) deficiency is a rare autosomal recessive metabolic disease that results in the decreased production of catecholamines. Standard treatment relies on combinations of levodopa and carbidopa, anticholinergic agents, serotonergic agonists, and monamine oxidase B inhibitors. Unfortunately, severely affected children often require escalating doses of medication and suffer from dyskinesias as well as significant on/off symptomatology. The authors present a case of medically intractable dopa-responsive dystonia in a 6-year-old boy whose condition significantly improved with bilateral subthalamic nucleus deep brain stimulation. This case is unique in its novel approach to tyrosine hydroxylase deficiency and the young age of the patient. PMID- 21631205 TI - Coagulation abnormalities in children undergoing epilepsy surgery. AB - OBJECT: Surgery is increasingly used to treat children with refractory epilepsy. Before surgery, the authors routinely evaluated the coagulation profile to identify coagulation abnormalities not established by personal and family history, physical examination, and routine screening tests. METHODS: Thirty-nine consecutive children undergoing testing prior to epilepsy surgery were prospectively evaluated. The authors evaluated a detailed hematological history and an elaborative hematological panel including complete blood count, hepatic panel, anticoagulant levels, coagulation profile (prothrombin time, partial thromboplastin time, international normalized ratio, fibrinogen, thrombin time, von Willebrand antigen, ristocetin cofactor, factor VIII, and individual factor assays when indicated) and platelet aggregation studies (in the presence of adenosine diphosphate, epinephrine, collagen, and ristocetin). Patient variables included tuberous sclerosis complex (TSC), age at epilepsy onset, age at surgery, seizure frequency, number and type of antiepileptic drugs, recent or present ketogenic diet, and use of selective serotonin reuptake inhibitors. RESULTS: Ten children (25.6%) had either coagulation or platelet function abnormalities. Abnormal coagulation was identified in 5 children, and abnormal platelet function was discovered in 6. A diagnosis of TSC was associated with a platelet function abnormality (p = 0.012), whereas children without TSC had a higher rate of coagulopathy (p = 0.041). None of the other characteristics reached statistical significance. In 2 patients (5.1%) with TSC and platelet aggregation abnormalities, the authors noted normal standard screening laboratory studies and an uneventful detailed personal and family history. One of these 2 patients developed a significant intraoperative bleeding complication. CONCLUSIONS: A preoperative screening with standard laboratory studies and detailed history may not be adequate to fully examine underlying coagulation abnormalities in children with refractory epilepsy. Platelet aggregation studies should be considered in patients with TSC. PMID- 21631206 TI - Temporoparietooccipital disconnection in children with intractable epilepsy. AB - OBJECT: Temporoparietooccipital (TPO) disconnection is described mainly in children with diffuse posterior quadrant lesions and concordant electroencephalography (EEG) findings. The authors report on 16 children who underwent TPO surgery, including 4 with no definite epileptogenic lesion and 8 with generalized electroclinical manifestations. METHODS: The authors conducted a retrospective review of clinical, neuropsychological, EEG, imaging, and histopathological data in 16 children with intractable epilepsy who underwent TPO disconnection and/or resection at their center between December 1998 and March 2010. RESULTS: Seizure onset occurred between the ages of 1 and 24 months, and TPO surgery was performed between the ages of 0.2 and 17 years. All children had refractory seizures, including epileptic spasms in 10 and tonic seizures in 7, and all had developmental delay. Twelve children had epileptogenic lesions on MR imaging, including 6 with posterior quadrant dysplasia. Four children had only subtle white matter signal change or unusual sulcation on MR imaging, associated with subtle but concordant EEG and functional imaging abnormalities. After a mean follow-up of 52 months (range 12-114 months), 9 children (56%) are seizure-free and 5 (31%) experienced seizure reduction of greater than 50%. Focal or regional background slowing on EEG was correlated with favorable seizure outcome. Five children showed developmental progress and 3 had acceleration in development following surgery. None of the children developed new motor deficits postoperatively. CONCLUSIONS: Temporoparietooccipital disconnection is an effective, motor-sparing epilepsy surgery procedure for selected children with refractory focal or generalized seizures with localization to the posterior quadrant on 1 side, with or without a discrete lesion on MR imaging. PMID- 21631207 TI - Vagal nerve stimulator infection: a lead-salvage protocol. AB - OBJECT: Vagal nerve stimulator (VNS) hardware infections are fraught with difficult management decisions. As with most implanted medical device-related infections, standard practice traditionally involves complete hardware removal, systemic antibiotic therapy, and subsequent reimplantation of the device. To avoid the potential morbidity of 2 repeat left carotid sheath surgical dissections, the authors have implemented a clinical protocol for managing VNS infections that involves generator removal and antibiotic therapy without lead removal. METHODS: A prospective, single-surgeon database was compared with hospital billing records to identify patients who underwent primary implantation or reimplantation of a VNS lead, generator, or both, from January 2001 to May 2010, at Oregon Health & Science University. From these records, the authors identified patients with VNS hardware infections and characterized their management, using a lead salvage protocol. RESULTS: In their review, the authors found a matching cohort of 206 children (age 3 months-17 years) who met the inclusion criteria. These children underwent 258 operations (including, in some children, multiple operations for generator end of life and/or lead malfunction). Six children experienced a single postimplantation infection (2.3% of the 258 operative cases), and no child experienced repeated infection. A lead-salvage protocol was used in 4 of 6 infected patients and was successful in 3 (75%), with clinical follow-up ranging from 10 months to 7.5 years. The fourth patient subsequently underwent lead removal and later reimplantation in standard fashion, with no adverse sequelae. CONCLUSIONS: Vagal nerve stimulator lead salvage is a safe and potentially advantageous strategy in the management of VNS-related infection. Further study is necessary to validate appropriate patient selection, success rates, and risks of this approach. PMID- 21631208 TI - Undeveloped semiconic posterosuperior ligament and susceptibility to avulsion of the C-7 spinal nerve in Erb palsy. AB - OBJECT: In Erb palsy, the C-7 spinal nerve has been found to be more subject to avulsion than the C-5 and C-6 spinal nerves. This study investigated the morphological and biomechanical characteristics of the semiconic posterosuperior ligaments (SPLs) at the C-5, C-6, and C-7 spinal nerves in neonates. METHODS: Twenty-four brachial plexuses from 12 fresh neonate cadavers were used in this study. In 12 brachial plexuses from 6 cadavers, the following studies were performed with respect to the SPLs at the C-5, C-6, and C-7 spinal nerves: gross observation of morphological and histological characteristics; measurement of length, thickness, and width; and a semiquantitative analysis of collagen. In the other 6 cadavers, biomechanical tension testing was performed bilaterally on the C5-7 SPLs to assess the tensile strength of the ligaments. RESULTS: The C5-7 spinal nerves are fixed to the transverse process through the SPL, a structure not observed at the C-8 and T-1 spinal nerves. Except for the width of the SPL insertion on the spinal nerve, which was found to increase gradually from C-5 to C-7, there was no statistically significant difference in the dimensions of the C 5, C-6, and C-7 SPLs. The sectional area percentage of collagen was 51% +/- 10% in SPLs for C-5, 51% +/- 11% for C-6, and 41% +/- 10% for C-7; and this percentage was significantly lower in SPLs for C-7 than for C-5 or C-6 (1-way ANOVA, F = 4.3, p = 0.02; Tukey honestly significant difference test, p = 0.04 and 0.04, respectively). Sharpey fibers were observed at the transverse process origin of the SPL at C-5 and C-6 but not at C-7. Biomechanical tension testing showed that the mean failure load was 6.6 +/- 0.9 N for the C-5 SPL, 6.4 +/- 1.0 N for the C-6 SPL, and 5.4 +/- 0.9 N for the C-7 SPL, and the failure load was significantly lower in SPLs at C-7 than in those at C-5 or C-6 (1-way ANOVA, F = 5.1, p = 0.01; Tukey honestly significant difference, p = 0.01 and 0.048, respectively). Nine of 12 C-7 SPLs failed at their origin from the transverse process, while only 4 of 12 C-5 SPLs and 3 of 12 C-6 SPLs failed at the origin site. CONCLUSIONS: These findings suggest that the lower density of collagen and absence of Sharpey fibers decrease the biomechanical properties of the C-7 SPL, and this may account for the higher frequency of avulsion of the C-7 spinal nerve (in comparison with the C-5 or C-6 nerve) in Erb palsy. PMID- 21631209 TI - Preoperative transfusion. PMID- 21631210 TI - Evidence-based review of primary and secondary ischemic stroke prevention in adults: a neurosurgical perspective. AB - In this paper, the authors' aim is to provide an evidence-based review of primary and secondary ischemic stroke prevention guidelines covering most of the common risk factors and stroke etiologies for the practicing neurosurgeon. The key to stroke prevention is in the identification and treatment of the major risk factors for stroke. These include hypertension, heart disease, diabetes mellitus, dyslipidemia, and tobacco smoking. An updated approach to secondary prevention of stroke in the setting of intracranial and extracranial large vessel atherosclerosis and cardioembolism is provided along with a brief overview of pertinent clinical trials. Novel pharmacological options for prevention of cardioembolic strokes, such as new alternatives to warfarin, are addressed with recommendations for interruption of therapy for elective surgical procedures. In addition, the authors have reviewed the anticoagulation guidelines and the risk of thromboembolic complications of such therapies in the perioperative period, which is an invaluable piece of information for neurosurgeons. Less common etiologies such as arterial dissections and patent foramen ovale are also briefly discussed. Finally, the authors have outlined the quality measures in the Medicare Physician Quality Reporting System and essential guidelines for Primary Stroke Center certification, which have implications for day-to-day neurosurgical practice. PMID- 21631211 TI - Endovascular stroke therapy: a single-center retrospective review. AB - OBJECT: Endovascular treatment of acute ischemic stroke delivers direct therapy at the site of an occluded cerebral artery and can be employed beyond the 3-4.5 hour window limit set for intravenous recombinant tissue plasminogen activator. In this paper, the authors report their experience with various endovascular therapies in acute ischemic stroke. METHODS: The authors conducted a retrospective review of their clinical database for acute ischemic stroke in large-vessel cerebral territories that underwent endovascular treatment between May 2005 and February 2009. Endovascular treatment was defined as pharmacological and/or mechanical intervention, angioplasty, stenting, or a combination of these methods. Admission National Institutes of Health Stroke Scale and the modified Rankin Scale scores were recorded. Thrombolysis in Myocardial Infarction (TIMI) scores of 0, 1, 2A, 2B, and 3 were used to define recanalization. RESULTS: Forty procedures were performed in 39 patients, with 1 patient having sequential bilateral strokes. Nine patients were lost to follow-up after discharge. Strokes in the carotid artery circulation occurred in 82.5% of cases, and those in the vertebral-basilar territory occurred in 17.5%. The Merci device was used in 22 (55%) of 40 procedures, and the Penumbra device in 9 (22.5%) of 40. Angioplasty was performed in 15 (37.5%) of 40 procedures, and intraarterial recombinant tissue plasminogen activator was administered in 23 (57.5%) of 40 procedures. In 23 (57.5%) of 40 cases, multiple recanalization methods were used. The recanalization rate for all methods was 60%. The recanalization rate from TIMI Score 0/1 occlusions was 71.4% (20 of 28). An estimated modified Rankin Scale score of <= 2 was obtained in 11 (36.7%) of 30 cases. The overall mortality rate was 26.7% (8 of 30). Intracerebral hemorrhage at 24 hours postprocedure was noted in 17 (42.5%) of 40 cases, 3 (7.5%) of which were symptomatic. CONCLUSIONS: The authors' institution performs endovascular stroke treatment with a safety and efficacy profile comparable to those of other major endovascular stroke therapy studies. Recanalization was associated with an improved clinical outcome. Protocols to maximize efficient triage of patients and better documentation of stroke treatments can assist in further studies. PMID- 21631212 TI - Endovascular stroke therapy. PMID- 21631213 TI - Hemodynamic instability following carotid artery stenting. AB - OBJECT: Postprocedural hypotension and bradycardia are important complications of carotid artery stenting (CAS) and are referred to as hemodynamic instability (HI). However, the incidence and impact of HI on the short-term prognosis of patients have been of a large debate. METHODS: Twenty-seven patients were selected based on NASCET criteria, and they underwent CAS between September 2008 and September 2009. Continuous electrocardiography monitoring and supine blood pressure (BP) monitoring were performed before and after stent deployment and on the following day to detect HI, defined as systolic BP lower than 90 mm Hg or a heart rate lower than 60 bpm. Patients were asked to perform a Valsalva maneuver before and after stent deployment. The Valsalva ratios (VRs) along with other demographic and procedural data were documented and compared between patients with and without incidence of HI. RESULTS: Seventeen patients (63%) developed HI after CAS. The degree of stenosis was found to have a significant correlation with occurrence of HI (p < 0.006). No other risk factor or demographic data showed any correlation with HI. The VRs were significantly lower in the HI group compared with the non-HI group, indicating a significant autonomic dysfunction (p < 0.003). During follow-up, 1 patient (4.3%) developed major stroke, and the remaining patients were symptom free. CONCLUSIONS: Hemodynamic instability occurs frequently after CAS, but it seems to be a benign phenomenon and does not increase the risk of mortality or morbidity in the short term. A VR at rest less than 1.10, baseline autonomic dysfunction, and degree of carotid artery stenosis can be used as measures for predicting HI after CAS. PMID- 21631214 TI - Contrast stasis on noncontrast computed tomography as a predictor of stroke postthrombolysis. AB - Multimodal endovascular intervention is becoming more commonplace for the acute intervention of ischemic stroke. Hyperdensity in a portion of the treated territory is a common finding on postthrombolytic noncontrast CT (NCCT), but its significance is poorly understood. The authors conducted a single-institution, retrospective chart review of patients who had intraarterial thrombolysis of the anterior circulation between 2010 and 2011 with evidence of hyperdensity on NCCT following recanalization. Eighteen patients had evidence of postoperative contrast stasis causing hyperdensity on NCCT. One hundred percent of the patients had MR imaging evidence of completed strokes postoperatively in the same distribution as the stasis. Stasis on NCCT after intervention had a sensitivity and specificity of 82% and 0% for predicting stroke, respectively. Furthermore, the positive predictive value was 100%. The presence of contrast stasis on postthrombolytic NCCT correlates well with stroke seen on subsequent MR imaging. PMID- 21631216 TI - Applications of stenting for intracranial atherosclerosis. AB - Intracranial atherosclerosis presents a therapeutic challenge to medical and surgical physicians alike. Despite maximal medical therapy, the stroke rate from this disease is still high, especially when arterial stenosis is severe and patients are symptomatic. Open surgical therapy has yet to be shown to be a more efficacious treatment than medical therapy alone, largely due to the relatively high rates of perioperative complications. Angioplasty has a similar fate, with the risk of periprocedural complications outweighing the overall benefit of treatment. With the advent of stents for use in intracranial vasculature, new hope has arisen for the treatment of intracranial atherosclerosis. The NEUROLINK system, the drug-eluting stents Taxus and Cypher, the flexible Wingspan stent, the Apollo stent, and the Pharos stent have all been used in various prospective and retrospective clinical studies with varying technical and clinical results. The authors' objective is to review and loosely compare the data presented for each of these stenting systems. While the Wingspan stent appears to have somewhat of an advantage with regard to technical success in comparison with the other stenting systems, the clinical follow-up time of its studies is too short to properly compare its complication rates with those of other stents. Before we continue to move forward with stenting for intracranial stenosis, a randomized prospective trial is ultimately needed to directly compare intracranial stenting to medical therapy. PMID- 21631215 TI - Symptomatic intracranial arterial disease: incidence, natural history, diagnosis, and management. AB - Symptomatic intracranial arterial disease is associated with a high rate of recurrent ischemic events. The management of this condition is controversial, with some advocating medical therapy as a sole means of treatment and others recommending endovascular therapy in addition to best medical management. In rare cases, surgical intervention is considered. A thorough review of the available literature was performed, and treatment recommendations based on these data are provided. PMID- 21631217 TI - Endovascular recanalization of symptomatic flow-limiting cervical carotid dissection in an isolated hemisphere. AB - OBJECT: Internal carotid artery dissection (ICAD) is a common cause of stroke in young patients, which may lead to major transient or permanent disability. Internal carotid artery dissection may occur spontaneously or after trauma and may present with a rapid neurological deterioration or with hemodynamic compromise and a delayed and unstable neurological deficit. Endovascular intervention using stent angioplasty can be used as an alternative to anticoagulation and open surgical therapy in this setting to restore blood flow through the affected carotid artery. METHODS: The authors present the cases of 2 patients with flow-limiting symptomatic ICAD leading to near-complete occlusion and without sufficient collateral supply. Both patients had isolated cerebral hemispheres without significant blood flow from the anterior or posterior communicating arteries. In both cases, the patients demonstrated blood pressure dependent subacute unstable neurological deficits as a result of the hemodynamic compromise resulting from the dissection. RESULTS: Both patients underwent careful microwire-based selection of the true lumen followed by confirmatory microinjection and subsequent exchange-length microwire-based recanalization using tandem telescoping endovascular stenting. In both cases the neurological state improved, and no permanent neurological deficit ensued. CONCLUSIONS: The treatment of ICAD may be difficult in patients with subacute unstable neurological deficits related to symptomatic hypoperfusion, especially in the setting of a hemodynamically isolated hemisphere. Anticoagulation alone may be insufficient in these patients. Although there is no widely accepted guideline for the treatment of ICAD, the authors recommend stent-mediated endovascular recanalization in cases of symptomatic flow-limiting hemodynamic compromise, especially in cases of an isolated hemisphere lacking sufficient communicating artery compensatory perfusion. PMID- 21631218 TI - Therapeutic hypothermia in acute ischemic stroke. AB - Induced hypothermia has been used for neuroprotection in cardiac and neurovascular procedures. Experimental and translational studies provide evidence for its utility in the treatment of ischemic cerebrovascular disease. Over the past decade, these principles have been applied to the clinical management of acute stroke. Varying induction methods, time windows, clinical indications, and adjuvant therapies have been studied. In this article the authors review the mechanisms and techniques for achieving therapeutic hypothermia in the setting of acute stroke, and they outline pertinent side effects and complications. The manuscript summarizes and examines the relevant clinical trials to date. Despite a reasonable amount of existing data, this review suggests that additional trials are warranted to define the optimal time window, temperature regimen, and precise clinical indications for induction of therapeutic hypothermia in the setting of acute stroke. PMID- 21631219 TI - Decompressive hemicraniectomy after malignant middle cerebral artery infarction: rationale and controversies. AB - Malignant middle cerebral artery stroke carries a very poor prognosis. Significant retrospective data support the hypothesis that decompressive hemicraniectomy decreases mortality rates due to this disease entity. Recently, 3 randomized controlled studies have been published and shed light on these issues and enhance the quality of evidence revolving around this procedure. In this review, the rationale, risks, benefits, and unanswered questions related to hemicraniectomy for acute ischemic stroke are reviewed with an emphasis on how 3 randomized trials have influenced knowledge on this life-saving yet controversial procedure. Further randomized studies are needed to clarify lingering questions regarding age indications and impact on quality of life. PMID- 21631220 TI - Determination of geographic variance in stroke prevalence using Internet search engine analytics. AB - OBJECT: Previous methods to determine stroke prevalence, such as nationwide surveys, are labor-intensive endeavors. Recent advances in search engine query analytics have led to a new metric for disease surveillance to evaluate symptomatic phenomenon, such as influenza. The authors hypothesized that the use of search engine query data can determine the prevalence of stroke. METHODS: The Google Insights for Search database was accessed to analyze anonymized search engine query data. The authors' search strategy utilized common search queries used when attempting either to identify the signs and symptoms of a stroke or to perform stroke education. The search logic was as follows: (stroke signs + stroke symptoms + mini stroke--heat) from January 1, 2005, to December 31, 2010. The relative number of searches performed (the interest level) for this search logic was established for all 50 states and the District of Columbia. A Pearson product moment correlation coefficient was calculated from the statespecific stroke prevalence data previously reported. RESULTS: Web search engine interest level was available for all 50 states and the District of Columbia over the time period for January 1, 2005-December 31, 2010. The interest level was highest in Alabama and Tennessee (100 and 96, respectively) and lowest in California and Virginia (58 and 53, respectively). The Pearson correlation coefficient (r) was calculated to be 0.47 (p = 0.0005, 2-tailed). CONCLUSIONS: Search engine query data analysis allows for the determination of relative stroke prevalence. Further investigation will reveal the reliability of this metric to determine temporal pattern analysis and prevalence in this and other symptomatic diseases. PMID- 21631221 TI - Evidence-based treatment of carotid artery stenosis. AB - Carotid atheromatous disease is an important cause of stroke. Carotid endarterectomy (CEA) is a well-established option for reducing the risk of subsequent stroke due to symptomatic stenosis (> 50%). With adequately low perioperative risk (< 3%) and sufficient life expectancy, CEA may be used for asymptomatic stenosis (> 60%). Recently, carotid angioplasty and stent placement (CAS) has emerged as an alternative revascularization technique. Trial design considerations are discussed in relation to trial results to provide an understanding of why some trials were considered positive whereas others were not. This review then addresses both the original randomized studies showing that CEA is superior to best medical management and the newer studies comparing the procedure to stent insertion in both symptomatic and asymptomatic populations. Additionally, recent population-based studies show that improvements in best medical management may be lowering the stroke risk for asymptomatic stenosis. Finally, the choice of revascularization technique is discussed with respect to symptom status. Based on current evidence, CAS should remain limited to specific indications. PMID- 21631222 TI - Moyamoya disease: a review of histopathology, biochemistry, and genetics. AB - OBJECT: Moyamoya disease (MMD) is a rare cerebrovascular disorder involving stenosis of the major vessels of the circle of Willis and proximal portions of its principal branches. Despite concerted investigation, the pathophysiology of the disorder has not been fully elucidated. Currently, the major proteins believed to play an active role in the pathogenesis include vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), transforming growth factor-beta1 (TGFbeta1), and granulocyte colony stimulating factor (G-CSF). In terms of the genetics, recent literature suggests a low penetrance autosomal dominant or polygenic mode of transmission involving chromosomes 3, 6, 8, 12, and 17 for familial MMD. This review summarizes the current knowledge on the histopathology, pathophysiology and genetics of MMD. METHODS: A PubMed/Medline systematic study of the literature was performed, from which 45 articles regarding MMD pathophysiology were identified and analyzed. CONCLUSIONS: Moyamoya disease is characterized by the intimal thickening and media attenuation of the proximal vessels of the circle of Willis as well as the development of an aberrant distal vascular network. The primary proteins that are currently implicated in the pathophysiology of MMD include VEGF, bFGF, HGF, TGFbeta1, and G-CSF. Furthermore, the current literature on familial MMD has pointed to a low penetrance autosomal dominant or polygenic mode of transmittance at loci on chromosomes 3, 6, 8, 12, and 17. PMID- 21631223 TI - Moyamoya disease: functional and neurocognitive outcomes in the pediatric and adult populations. AB - OBJECT: Moyamoya disease is an occlusive cerebrovascular disorder commonly resulting in neurocognitive impairment. The cognitive outcome parameters commonly affected are intelligence, memory, executive function, and quality of life. In this paper, the authors review the existing literature on cognitive and clinical outcomes in adult and pediatric moyamoya populations separately. METHODS: A systematic review of the cognitive and clinical outcome literature was performed using the PubMed/MEDLINE database. Outcomes data were contrasted between adult and pediatric populations. RESULTS: Intelligence is the main cognitive outcome parameter affected in pediatric patients with moyamoya disease, whereas adults most commonly suffer from executive function impairment. Memory has not been studied sufficiently in pediatric patients, and its dysfunction in the adult population remains controversial. Quality of life has not been studied appropriately in either population. Surgical revascularization is the only beneficial treatment option, and a combination of direct and indirect bypass techniques has shown benefit, but the impact on the above-mentioned parameters has not been sufficiently elucidated. CONCLUSIONS: Moyamoya disease affects the cognition and daily function in pediatric patients to a greater extent than in adult patients. Due to the rarity of the disease, there is a distinct lack of high-level evidence regarding cognitive and clinical outcomes. PMID- 21631224 TI - State of the art in managing nontraumatic intracerebral hemorrhage. AB - Nontraumatic intracerebral hemorrhage constitutes a major public health problem worldwide. Intracerebral hemorrhage leads to a high rate of morbidity and mortality. To date, no medical or surgical trials have clearly attested to the benefit of a particular therapy. The aim of this review was to summarize the best evidence for management decision-making in intracerebral hemorrhage. PMID- 21631225 TI - The relationships between endothelial nitric oxide synthase polymorphisms and the formation of intracranial aneurysms in the Korean population. AB - OBJECT: Some genetic factors are known to be associated with the formation of cerebral aneurysms in the Caucasian population. One of these factors is endothelial nitric oxide synthase (eNOS) gene polymorphisms. Endothelial nitric oxide synthase genes encode eNOS, which synthesizes NO from l-arginine. There continues to be controversy about the relationships between eNOS gene polymorphisms and the formation of intracranial aneurysms. In this study, the authors evaluated these relationships in the Korean population. METHODS: Three eNOS polymorphisms (eNOS 27VNTR, T786C, and G894T) were genotyped in 96 patients with ruptured aneurysms, 53 patients with unruptured aneurysms, and in 121 volunteers via polymerase chain reaction-restriction fragment length polymorphism analysis. RESULTS: The mean ages of the patients and healthy volunteers were 52.9 +/- 12.3 years and 55.2 +/- 9.1 years, respectively. The patient group was composed of 56 men and 93 women, and the healthy volunteer group was composed of 46 men and 75 women. Only the incidence of smoking history was significantly higher in the patient group than in the control group (p = 0.001). The genotypic frequencies for the 3 eNOS gene polymorphisms were in agreement with those predicted by Hardy-Weinberg equilibrium. There were no significant associations between the eNOS recessive models and the formation of an aneurysm. The authors found no genotypic differences between similar races among patients with aneurysms. CONCLUSIONS: The present study shows that eNOS 27VNTR, T786C, and G894T polymorphisms cannot be used as indicators of the formation of intracranial aneurysms in Korean patients. To confirm these findings an additional analyses might need to be performed using a larger sample size. There were no differences in the genotypic distributions and allelic frequencies between similar races among patients with aneurysms, which were the same in previously reported normal populations. PMID- 21631226 TI - The natural history of intracranial cavernous malformations. AB - Literature reports on the natural history of cerebral cavernous malformations (CMs) are numerous, with considerable variability in lesion epidemiology, hemorrhage rates, and risk factors for hemorrhage. In this review, the authors performed a meta-analysis of 11 natural history studies. The overall male-to female ratio was 1:1, and the mean age at presentation was 30.6 years. Overall, 37% of patients presented with seizures, 36% with hemorrhage, 23% with headaches, 22% with focal neurological deficits, and 10% were asymptomatic. Some patients had more than one symptom. Seizure presentation was most prevalent among supratentorial CMs, while focal neurological deficits were common in patients with infratentorial CMs. By location, CMs were in the cerebral hemispheres (66%), brainstem (18%), basal ganglia or thalamus (8%), cerebellum (6%), and other (2.5% [combined supra- and infratentorial, callosal or insular]). Overall, 19% of patients harbored multiple intracranial CMs, and 9% had radiographically apparent associated developmental venous anomalies. An overall annual hemorrhage rate of 2.4% per patient-year (range 1.6%-3.1%) was identified across 3 studies. Prior hemorrhage and female sex were risk factors for bleeding, while CM size and multiplicity did not affect hemorrhage rates. Although not impacting the hemorrhage rate itself, deep location was a risk factor for increased clinical aggressiveness. PMID- 21631227 TI - Re: Evidence-based treatment of carotid stenosis: is the evidence strong enough? PMID- 21631228 TI - Utility of CT perfusion scanning in patient selection for acute stroke intervention: experience at University at Buffalo Neurosurgery-Millard Fillmore Gates Circle Hospital. AB - Computed tomography perfusion scanning generates physiological flow parameters of the brain parenchyma, allowing differentiation of ischemic penumbra and core infarct. Perfusion maps, along with the National Institutes of Health Stroke Scale score, are used as the bases for endovascular stroke intervention at the authors' institute, regardless of the time interval from stroke onset. With case examples, the authors illustrate their perfusion-based imaging guidelines in patient selection for endovascular treatment in the setting of acute stroke. PMID- 21631229 TI - Advanced neuroimaging in acute ischemic stroke: extending the time window for treatment. AB - Early treatment of ischemic stroke with thrombolytics is associated with improved outcomes, but few stroke patients receive thrombolytic treatment in part due to the 3-hour time window. Advances in neuroimaging may help to aid in the selection of patients who may still benefit from thrombolytic treatment beyond conventional time-based guidelines. In this article the authors review the available literature in support of using advanced neuroimaging to select patients for treatment beyond the 3-hour time window cutoff and explore potential applications and limitations of perfusion imaging in the treatment of acute ischemic stroke. PMID- 21631230 TI - Computed tomography perfusion-based selection of patients for endovascular recanalization. AB - Intravenous and intraarterial recombinant tissue plasminogen activator remains underutilized in the treatment of acute ischemic stroke, largely due to strict adherence to the concept of the therapeutic time window for administration. Recent efforts to expand the number of patients eligible for thrombolysis have been mirrored by an evolution in endovascular recanalization technology and techniques. As a result, there is a growing need to establish efficient and reliable means by which to select candidates for endovascular intervention beyond the traditional criteria of time from symptom onset. Perfusion imaging techniques, particularly CT perfusion used in combination with CT angiography, represent an increasingly recognized means by which to identify those patients who stand to benefit most from endovascular recanalization. Additionally, CT perfusion and CT angiography appear to provide sufficient data by which to exclude patients in whom there is little chance of neurological recovery or a substantial risk of postprocedure symptomatic intracranial hemorrhage. The authors review the current literature as it pertains to the limitations of time based selection of patients for intervention, the increasing utilization of endovascular therapy, and the development of a CT perfusion-based selection of acute stroke patients for endovascular recanalization. Future endeavors must prospectively evaluate the utility and safety of CT perfusion-based selection of candidates for endovascular intervention. PMID- 21631231 TI - Improving patient selection for endovascular treatment of acute cerebral ischemia: a review of the literature and an external validation of the Houston IAT and THRIVE predictive scoring systems. AB - Outcome after intraarterial therapy (IAT) for acute ischemic stroke remains variable, suggesting that improved patient selection is needed to better identify patients likely to benefit from treatment. The authors evaluate the predictive accuracies of the Houston IAT (HIAT) and the Totaled Health Risks in Vascular Events (THRIVE) scores in an independent cohort and review the existing literature detailing additional predictive factors to be used in patient selection for IAT. They reviewed their center's endovascular records from January 2004 to July 2010 and identified patients who had acute ischemic stroke and underwent IAT. They calculated individual HIAT and THRIVE scores using patient age, admission National Institutes of Health Stroke Scale (NIHSS) score, admission glucose level, and medical history. The scores' predictive accuracies for good outcome (discharge modified Rankin Scale score <= 3) were analyzed using receiver operating characteristics analysis. The THRIVE score predicts poor outcome after IAT with reasonable accuracy and may perform better than the HIAT score. Nevertheless, both measures may have significant clinical utility; further validation in larger cohorts that accounts for differences in patient demographic characteristics, variation in time-to-treatment, and center preferences with respect to IAT modalities is needed. Additional patient predictive factors have been reported but not yet incorporated into predictive scales; the authors suggest the need for additional data analysis to determine the independent predictive value of patient admission NIHSS score, age, admission hyperglycemia, patient comorbidities, thrombus burden, collateral flow, time to treatment, and baseline neuroimaging findings. PMID- 21631232 TI - Sources of variability in computed tomography perfusion: implications for acute stroke management. AB - OBJECT: Although dynamic, first-pass cerebral CT perfusion is used in the evaluation of acute ischemic stroke, a lack of standardization restricts the value of this imaging modality in clinical decision-making. The purpose of this study was to comprehensively review the reported sources of variability and error in cerebral CT perfusion results. METHODS: A systematic literature review was conducted, 120 articles were reviewed, and 23 published original research articles were included. Sources of variability and error were thematically categorized and presented within the context of the 3 stages of a typical CT perfusion study: data acquisition, postprocessing, and results interpretation. RESULTS: Seven factors that caused variability were identified and described in detail: 1) contrast media, the iodinated compound injected intravascularly to permit imaging of the cerebral vessels; 2) data acquisition rate, the number of images obtained by CT scan per unit time; 3) user inputs, the subjective selections that operators make; 4) observer variation, the failure of operators to repeatedly measure a perfusion parameter with precision; 5) software operational mode, manual, semiautomatic, or automatic; 6) software design, the mathematical algorithms used to perform postprocessing; and 7) value type, absolute versus relative values. CONCLUSIONS: Standardization at all 3 stages of the CT perfusion study cycle is warranted. At present, caution should be exercised when interpreting CT perfusion results as these values may vary considerably depending on a variety of factors. Future research is needed to define the role of CT perfusion in clinical decision-making for acute stroke patients and to determine the clinically acceptable limits of variability in CT perfusion results. PMID- 21631233 TI - Computed tomography perfusion. PMID- 21631235 TI - Aggressive surgical intervention in end-stage renal disease patients with spinal epidural abscess. AB - BACKGROUND: Despite advances in imaging, as well as antibiotic and surgical treatment, spinal epidural abscess (SEA) remains a challenging problem in end stage renal disease (ESRD) patients. This investigation assesses the influence of ESRD on clinical manifestations, complications, and outcomes in patients with SEA. METHODS: This study retrospectively reviewed medical records of 41 patients with SEA treated during 2003-2006. The patients comprised two groups: group I (patients with ESRD) and group II (patients without ESRD). Patient characteristics, including age, gender, comorbidities, clinical presentations, laboratory data, locations of epidural abscess, and outcome, were recorded and compared. RESULTS: The final sample comprised 41 patients. The mean age of the subjects was 62 + 12 years. The sample included 12 patients with ESRD and 29 without ESRD. The development of symptoms was similar for both groups. Group I patients displayed higher serum erythrocyte sedimentation rate (ESR) (108 + 26 vs. 81 + 31 mm/h, p = 0.014) and lower serum hematocrit (27.1 + 4.3 vs. 33.7 + 5.1%, p < 0.001) than group II patients. Overall survival at 3 months was 88.9% and 93.1% for patients in groups I and II, respectively (p = 0.876). Meanwhile, patients that underwent surgical decompression of epidural abscess received more surgeries than group I patients (1.08 vs. 0.55, p = 0.086), although the difference was not significant. CONCLUSIONS: ESRD is a risk factor for repeat surgery in patients with SEA. This investigation suggests that ESRD patients with SEA may require aggressive surgical intervention despite ESRD not affecting their long-term prognosis. PMID- 21631236 TI - An autopsy case of mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) with intestinal bleeding in chronic renal failure. AB - A 50-year-old man who underwent hemodialysis (HD) at local outpatient HD center due to end-stage renal disease (ESRD) was transferred to our hospital because of pneumonia. He had severe emaciation and past history of congestive heart failure. Presenting symptoms almost consistently involved difficulty in hearing and recurrent attacks of migraine-like headaches. He was diagnosed with dilated cardiomyopathy, showing diastolic mechanical dyssynchrony by tissue Doppler echocardiography. On the day of death, he had hematemesis and hemorrhagic shock. Autopsy revealed perforation of duodenum, and genetic analysis using mitochondrial DNA from cardiac muscle and iliopsoas muscle revealed a 3243A > G mutation in the mitochondrial tRNA(Leu(UUR)) gene, which is related to mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS). Multiple organ failure due to the mutation of mitochondrial DNA with gastrointestinal bleeding is not a common. PMID- 21631237 TI - Evidence of in vitro differential secretion of human beta-defensins-1, -2, and -3 after selective exposure to Streptococcus agalactiae in human fetal membranes. AB - OBJECTIVE: The aim of this work was to characterize the individual contribution of the amnion (AMN) and choriodecidua (CHD) regions to the secretion of human beta defensins (HBD)-1, -2, and -3, after stimulation with Streptococcus agalactiae. METHODS: Full-thickness membranes were mounted on a Transwell device, constituted by two independent chambers; 1 * 10(6) CFU/ml of S. agalactiae were added to either the AMN or CHD face or to both. Secretion profiles of HBD-1, HBD 2, and HBD-3 to the culture medium were quantified by enzyme-linked immunosorbent sandwich assay (ELISA). RESULTS: Secretion profile of HBD-1 remained without significant changes; HBD-2 secretion level by the CHD increased 2.0 (2.73 +/- 0.19 pg/MUg) and 2.6 (3.62 +/- 0.60 pg/MUg) times when the stimulus was applied only to the CHD region and simultaneously to both compartments, respectively. The bacterial stimulation in the AMN induced a 2.0 times (2.06 +/- 0.29 pg/MUg) increase in this region. HBD-3 secretion level increased significantly in the CHD (15.65 +/- 2.68 pg/MUg) and the AMN (14.94 +/- 1.85 pg/MUg) only when both regions were stimulated simultaneously. CONCLUSION: The stimulation of human fetal membranes with S. agalactiae induced a differential and tissue-specific profile of HBD-1, HBD-2, and HBD-3 secretion. PMID- 21631238 TI - Genetic variation in the HSD3B1 gene and recurrent spontaneous abortions. AB - OBJECTIVE: HSD3B1 gene encodes the 3beta-hydroxysteroid deydrogenases/isomerase (3beta-HSD) enzyme, which plays a crucial role in the biosynthesis of all hormonal steroids. The aim of this study was to examine the potential impact of a T -> C substitution at codon Leu(338) of HSD3B1 gene on pregnancy outcome. METHODS: In this prospective case-control study, 162 patients and 139 healthy controls were investigated for the possible association between the HSD3B1 T/C polymorphism and the risk of recurrent spontaneous abortions (RSA). The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used in order to genotype the subjects. RESULTS: The frequencies of TT, TC, and CC genotypes were 0.20, 0.51, and 0.29, respectively, in the patient group and 0.20, 0.45, and 0.35, respectively, in the control group. The allele frequencies were 0.456 and 0.428 for T allele for the patient group and control group, respectively and 0.543 and 0.572 for C allele for the patient and control group, respectively. The data between the two groups were analyzed by chi-square test or Fisher's exact test. Our results showed that there are no significant differences in genotype (P = 0.56) or in allele frequencies (P = 0.51) between the patient and the control group. CONCLUSION: The HSD3B1 T/C polymorphism cannot be used as genetic marker for the risk for RSA in our Caucasian population. PMID- 21631239 TI - Treatment of gestational diabetes mellitus: glyburide compared to subcutaneous insulin therapy and associated perinatal outcomes. AB - OBJECTIVE: To examine perinatal outcomes in women with gestational diabetes mellitus treated with glyburide compared to insulin injections. STUDY DESIGN: This is a retrospective cohort study of women diagnosed with gestational diabetes mellitus (GDM) who required pharmaceutical therapy and were enrolled in the Sweet Success California Diabetes and Pregnancy Program between 2001 and 2004, a California state-wide program. Women managed with glyburide were compared to women treated with insulin injections. Perinatal outcomes were compared using chi square test and multivariable logistic regression models; statistical significance was indicated by p < 0.05 and 95% confidence intervals (CI). RESULTS: Among the 10,682 women with GDM who required medical therapy and met study criteria, 2073 (19.4%) received glyburide and 8609 (80.6%) received subcutaneous insulin injections. Compared to insulin therapy and controlling for confounders, oral hypoglycemic treatment was associated with increased risk of birthweight >4000 g (aOR = 1.29; 95% CI [1.03-1.64]), and admission to the intensive care nursery (aOR = 1.46 [1.07-2.00]). CONCLUSION: Neonates born to women with gestational diabetes managed on glyburide, and were more likely to be macrosomic and to be admitted to the intensive care unit compared to those treated with insulin injections. These findings should be examined in a large, prospective trial. PMID- 21631240 TI - Mannose-binding lectin serum levels in neonatal sepsis and septic shock. AB - OBJECTIVE: To evaluate mannose-binding lectin (MBL) serum levels as a marker for predicting sepsis, septic shock, and their outcomes in neonates. PATIENTS AND METHODS: A prospective study was conducted on 62 neonates (27 preterm and 35 full term) with culture-proven sepsis and 35 controls. Serum levels of MBL were measured by immunoassay. RESULTS: Of 62 infants with positive blood cultures (Gram-negative = 44 and Gram-positive = 18), 11 infants had severe sepsis and 6 neonates developed septic shock. MBL levels were significantly lower in infants with sepsis than in control group (0.39 +/- 0.07 vs. 1.34 +/- 0.03 MUg/ml; p < 0.001). The lowest MBL levels were detected in those infants with septic shock, particularly those who died (p < 0.05). MBL had high sensitivity (96.7), specificity (97.1), positive (98.3), and negative (94.4) predictive values to detect sepsis. CONCLUSION: Low MBL serum levels could be considered as sensitive and specific marker for predicting sepsis, septic shock, and their clinical outcomes in newborn infants. PMID- 21631242 TI - Long-term complications of assisted reproductive technologies. AB - Since the birth of the first baby through in vitro fertilisation over 30 years ago, there has been a rapid increase in the number of women using assisted reproductive technologies. New techniques are constantly being introduced and are evolving rapidly. Understanding the long-term impact of these treatments, which have been linked with several potential complications affecting the mother and the child, poses a constant challenge for the scientific community. The main complications discussed in this review include the rare but serious risk of thromboembolic disease particularly in the upper half of the body and a higher risk of pregnancy complications including abnormal placentation, miscarriage, gestational diabetes and hypertensive disorders. A link between assisted conception and a number of genital cancers particularly of the breast, ovary and endometrium has also been suggested, but the evidence is currently inconclusive. A number of foetal and neonatal complications including imprinting disorders, low birth weight, congenital malformations and growth disorders are also discussed. PMID- 21631241 TI - Zinc-finger nucleases for somatic gene therapy: the next frontier. AB - Zinc-finger nucleases (ZFNs) are a powerful tool that can be used to edit the human genome ad libitum. The technology has experienced remarkable development in the last few years with regard to both the target site specificity and the engineering platforms used to generate zinc-finger proteins. As a result, two phase I clinical trials aimed at knocking out the CCR5 receptor in T cells isolated from HIV patients to protect these lymphocytes from infection with the virus have been initiated. Moreover, ZFNs have been successfully employed to knockout or correct disease-related genes in human stem cells, including hematopoietic precursor cells and induced pluripotent stem cells. Targeted genome engineering approaches in multipotent and pluripotent stem cells hold great promise for future strategies geared toward correcting inborn mutations for personalized cell replacement therapies. This review describes how ZFNs have been applied to models of gene therapy, discusses the opportunities and the risks associated with this novel technology, and suggests future directions for their safe application in therapeutic genome engineering. PMID- 21631243 TI - Quality manual project. PMID- 21631244 TI - The use of single embryo transfer to reduce the incidence of twins: Implications and questions for practice from the 'towardSET?' project. AB - In vitro fertilisation treatments where multiple embryos are transferred are associated with high multiple birth rates leading to a corresponding high infant morbidity. Here we review the results from a multidisciplinary project which aimed to combine state of the art statistical modelling of routine clinical data with consideration of patient perspectives to explore options for reducing multiple birth incidence by increased use of single embryo transfer (SET). Modelling was based on a large multicentre cohort, supplemented by analysis of HFEA register data. Patient perspectives were explored in qualitative interviews and focus groups. The data confirm the reduction of around one-third in the chance of a live birth for any couple in moving from double embryo transfer (DET) to SET in a fresh cycle. This can be somewhat offset by appropriate patient and cycle selection for SET, with many suggested schemes performing similarly, although many patients perceive such selection as unfair. If we take a complete cycle perspective, and consider the transfer of all good-quality embryos with cryopreservation then it is possible for SET to match or even outperform DET. However, the additional treatment cycles are seen by patients as physically and emotionally burdensome. Such treatments will require optimisation of embryo freezing policies and a number of options are explored. PMID- 21631245 TI - Treatment decision making in anonymous donor egg in-vitro fertilisation: a qualitative study of childless women and women with genetically related children. AB - The aim of this study was to analyse donor egg recipients' anticipatory concerns and feelings and to consider how these relate to their decision to enter into treatment. The women were divided into two groups: those who were childless and those with a genetically related child or children. In-depth interviews were carried out with seven women on the waiting list for the egg donation treatment programme at the Lister Hospital in London. The participants with children expressed reservations and anxiety about proceeding with treatment and this created 'a state of dilemma' but the possibility of treatment led to a pressure to proceed. In contrast, the prospect of treatment for the childless participants gave rise to feelings of excitement and hope of finally having a baby and their quest took on the form of a personal mission and they expressed very few reservations about the treatment. This was a small qualitative study but in terms of egg donation research, the distinction seen here between women seeking to be first time mothers and those attempting to build a new family or complete an existing one, warrants further attention and has implications for pre-treatment counselling. PMID- 21631246 TI - Reconsidering the number of offspring per gamete donor in the Dutch open-identity system. AB - The introduction of legislation in the Netherlands in 2004 enabling donor offspring to identify and make contact with their donors has led to a need to reconsider the number of offspring that an individual semen donor may produce. To this end, we made a survey on the limits for offspring per donor in 29 different countries, distinguishing between systems with anonymous and open-identity sperm donation. We counted donations as individual offspring conceived, or as the number of women/families helped, by a single donor. The interests of the various participants in gamete donation in open-identity and anonymous systems are considered with some of the basic assumptions and interactions between the different parties that might be used to formulate reasonable guidelines. A number of recommendations are made. PMID- 21631247 TI - Physical symptoms throughout IVF cycles. AB - The physical impact of in vitro fertilisation (IVF) is not well explored. The aim of this prospective controlled longitudinal study was to examine the impact of 11 symptoms during an entire long down regulation (LDR) cycle and to determine the effect of the mean recombinant follicle stimulating hormone (RecFSH) dose and ovarian responsiveness of patients and outcome had on reported symptoms. The severity of symptoms was measured using a daily questionnaire to determine a total summary score (TSS) for each symptom, a summary symptom score (SSS) based on all TSS and a mean daily severity for each symptom. Outcome was determined by beta human chorionic gonadotrophin (beta-hCG) test result. 79.1% of women undertaking LDR IVF cycles had significant physical symptoms. Treatment symptom severity peaked at oocyte retrieval, with prominent symptoms including bloating, abdominal pain/cramps and fatigue. No relationship was found between outcome, RecFSH dose and ovarian responsiveness with reported symptoms. In conclusion, IVF places a real physical burden on women should be encouraged to reduce undertaking LDR cycles. The impact of the symptoms, particularly the increase in severity leading up to day of oocyte retrieval suggests that women may be warned to reduce stress and activities during this time. PMID- 21631250 TI - Earth as an extrasolar planet: Earth model validation using EPOXI earth observations. AB - The EPOXI Discovery Mission of Opportunity reused the Deep Impact flyby spacecraft to obtain spatially and temporally resolved visible photometric and moderate resolution near-infrared (NIR) spectroscopic observations of Earth. These remote observations provide a rigorous validation of whole-disk Earth model simulations used to better understand remotely detectable extrasolar planet characteristics. We have used these data to upgrade, correct, and validate the NASA Astrobiology Institute's Virtual Planetary Laboratory three-dimensional line by-line, multiple-scattering spectral Earth model. This comprehensive model now includes specular reflectance from the ocean and explicitly includes atmospheric effects such as Rayleigh scattering, gas absorption, and temperature structure. We have used this model to generate spatially and temporally resolved synthetic spectra and images of Earth for the dates of EPOXI observation. Model parameters were varied to yield an optimum fit to the data. We found that a minimum spatial resolution of ~100 pixels on the visible disk, and four categories of water clouds, which were defined by using observed cloud positions and optical thicknesses, were needed to yield acceptable fits. The validated model provides a simultaneous fit to Earth's lightcurve, absolute brightness, and spectral data, with a root-mean-square (RMS) error of typically less than 3% for the multiwavelength lightcurves and residuals of ~10% for the absolute brightness throughout the visible and NIR spectral range. We have extended our validation into the mid-infrared by comparing the model to high spectral resolution observations of Earth from the Atmospheric Infrared Sounder, obtaining a fit with residuals of ~7% and brightness temperature errors of less than 1 K in the atmospheric window. For the purpose of understanding the observable characteristics of the distant Earth at arbitrary viewing geometry and observing cadence, our validated forward model can be used to simulate Earth's time dependent brightness and spectral properties for wavelengths from the far ultraviolet to the far infrared. Key Words: Astrobiology-Extrasolar terrestrial planets-Habitability-Planetary science-Radiative transfer. Astrobiology 11, 393 408. PMID- 21631251 TI - Relationship of lower breastfeeding score and problems in infancy. AB - BACKGROUND AND METHODS: We undertook a descriptive study in 2-month-old healthy infants to determine the factors that affect breastfeeding score. Mother's breastfeeding was evaluated and scored according to the World Health Organization/UNICEF B-R-E-A-S-T Feeding Observation Form. RESULTS: The breastfeeding score (BFS) was higher in female than male babies (p=0.005). The babies with regurgitation had lower BFS than the babies without (p=0.016). The BFS was lower in babies who had repeated, without cause, inconsolable crying than in those without such crying (p<0.004). When the crying was problematic for the family, BFS was lower (p=0.028). Babies who had another sibling with a history of colic had a lower BFS (p=0.038). A low BFS was associated with short duration of night sleeping (p=0.032). CONCLUSIONS: A decreased BFS may be a risk factor or indicator for infant crying, regurgitation, and short sleeping duration. As a result, tracking the BFS and appropriate breastfeeding intervention during the newborn period may assist in decreasing the frequency of regurgitation and infant crying. PMID- 21631252 TI - Male influence on infant feeding in rural Guatemala and implications for child nutrition interventions. AB - BACKGROUND AND AIMS: Guatemala has one of the highest rates of child stunting in the world, which especially impacts rural indigenous agricultural communities. Despite decades of intensive nutrition research and interventions, only rarely have nutrition programs successfully lowered the rate of stunting in these settings. The bulk of nutritional interventions in Guatemala are targeted at the education of female caregivers. However, women's ability to implement best practices in infant breastfeeding and complementary feeding are often constrained by external factors. This study evaluated the knowledge, beliefs, and practices of female caregivers, as well as the attitudes of fathers, toward breastfeeding and infant feeding in a rural Guatemalan village. METHODS: Clinical work, participant-observation, surveys, interviews, and focus groups were conducted in a rural Guatemalan village in conjunction with a child feeding program from August 2008 to January 2011. RESULTS: Male employment status, mental health, and attitudes towards child rearing and parenting responsibilities are often principal factors in infant growth failure. CONCLUSIONS: Successful child feeding programs must include educational elements for men and should consider structural elements that provide a safety net for unexpected changes in domestic finances. PMID- 21631253 TI - Factors that influence breastmilk and a tale from west virginia. PMID- 21631254 TI - ABM clinical protocol #10: breastfeeding the late preterm infant (34(0/7) to 36(6/7) weeks gestation) (first revision June 2011). AB - A central goal of The Academy of Breastfeeding Medicine is the development of clinical protocols for managing common medical problems that may impact breastfeeding success. These protocols serve only as guidelines for the care of breastfeeding mothers and infants and do not delineate an exclusive course of treatment or serve as standards of medical care. Variations in treatment may be appropriate according to the needs of an individual patient. These guidelines are not intended to be all-inclusive, but to provide a basic framework for physician education regarding breastfeeding. PMID- 21631257 TI - Third-track pathology: in unambiguous support of the Banbury Conference report. PMID- 21631258 TI - Citius, altius, fortius-self-assessment in pathology and laboratory medicine: College of American Pathologists Self-assessment Module Committee. PMID- 21631259 TI - Synoptic reporting for central nervous system tumors: has its time come? PMID- 21631260 TI - Where and by whom should gastric cancer HER2/neu status be assessed?: lessons from breast cancer. PMID- 21631261 TI - The 9th spring seminar of the Korean Pathologists Association of North America. PMID- 21631262 TI - Four molecular subtypes of colorectal cancer and their precursor lesions. AB - CONTEXT: In addition to chromosomal instability and microsatellite instability (MSI), a third pathway, epigenetic instability, has been implicated in progression to colorectal carcinogenesis. CpG island methylator phenotype (CIMP) refers to a subset of colorectal cancers (CRCs) that occur through the epigenetic instability pathway and that are characterized by widespread hypermethylation of promoter CpG island loci, resulting in the inactivation of several tumor suppressor genes or tumor-related genes. Colorectal cancers can be classified into 4 molecular subtypes according to their CIMP and MSI statuses: CIMP+/MSI+, CIMP+/MSI-, CIMP-/MSI+, and CIMP-/MSI-. There are differences between Western (United States and European Union) and Eastern (Korea and China) populations in the number of CRCs that are MSI+, and in the number of MSI+ CRCs that are CIMP+. OBJECTIVE: To review the clinicopathologic and molecular features of the 4 molecular subtypes of CRCs and their precursor lesions, and to emphasize geographic differences in CRCs between Eastern and Western populations. DATA SOURCES: This article is based on the author's own experimental data and a literature review of relevant articles indexed in PubMed (US National Library of Medicine). CONCLUSION: The 4 molecular subtypes of CRC that are defined by their CIMP and MSI statuses are characterized by their own distinct clinicopathologic and molecular features and precursor lesions. In particular, the clinicopathologic features of MSI+ CRCs differ depending on the CIMP status. Further understanding of the heterogeneity in CRC molecular pathways may help to explain the diverse morphologic features of CRCs. PMID- 21631263 TI - Update on precursor and early lesions of hepatocellular carcinomas. AB - CONTEXT: There is increasing evidence to support a multistep model of the process of human hepatocarcinogenesis. Precursor lesions are characterized by the appearance of dysplastic lesions in the form of microscopic dysplastic foci and macroscopic dysplastic nodules. There are 2 types of small hepatocellular carcinoma (HCC) (<=2 cm in diameter): (1) early HCC with an indistinct margin and (2) progressed HCC with a distinct margin. Pathologic diagnostic criteria for early HCC have recently been set up based on a consensus between Eastern and Western pathologists. OBJECTIVE: To review the nomenclature, pathology, and biomarkers of precursor and early lesions of HCC. DATA SOURCES: Literature review and illustrations from case materials were used. CONCLUSIONS: Dysplastic foci are composed of large and small cell changes. Small cell change is considered to be a more advanced precursor lesion than large cell change, and large cell change is a rather heterogeneous lesion that may represent both reactive change and true dysplasia. Dysplastic nodules can be categorized as low or high grade according to the degree of atypia. High-grade dysplastic nodules have been reported to show molecular changes similar to HCC and have a high risk of malignant transformation. Early HCC, which may correspond to microinvasive carcinomas of other organs, is a well-differentiated HCC, and differential diagnosis between early HCC and high-grade dysplastic nodule is difficult. Identification of stromal invasion and application of a panel of markers (glypican-3, heat shock protein 70, and glutamine synthetase) is helpful for diagnosis of early HCC. Detection of precursor lesions of HCC is important in recognizing patients with higher risk of developing HCC, and diagnosis of early HCC can improve patient survival by allowing for early and adequate treatment. PMID- 21631265 TI - The College of American Pathologists and National Society for Histotechnology workload study. AB - Limited data exist in regard to productivity and staffing in the anatomic pathology laboratory. In 2004, the National Society for Histotechnology (NSH) conducted a pilot study to examine productivity and staffing in the histology laboratory. After review of the data, The College of American Pathologists (CAP)/NSH Histotechnology Committee concluded that a larger survey was required to further address and expand on the pilot study findings. In 2007, a total of 2674 surveys were sent out to North American laboratories. From the responses, comparisons of laboratory demographics and productivity were examined by institution type and workload volume. Productivity was measured as the number of paraffin-embedded tissue blocks processed per full-time equivalent per year. This manuscript presents and discusses the data collected from the CAP/NSH Workload Study. PMID- 21631264 TI - Molecular signatures of pancreatic cancer. AB - CONTEXT: The introduction of genome- and epigenome-wide screening techniques has dramatically improved our understanding of the molecular mechanisms underlying the development of pancreatic cancer. There are now 3 recognized histologic precursors of pancreatic cancer: pancreatic intraepithelial neoplasia, intraductal papillary mucinous neoplasm, and mucinous cystic neoplasm. Each of these precursor lesions is associated with specific molecular alterations. OBJECTIVE: To understand the molecular characteristics of pancreatic ductal adenocarcinoma and its precursor lesions. DATA SOURCES: PubMed (US National Library of Medicine). CONCLUSIONS: In this review, we briefly summarize recent research findings on the genetics and epigenetics of pancreatic cancer. In addition, we characterize these molecular alterations in the context of the histologic subtypes of pancreatic cancer. PMID- 21631266 TI - Clinical implications of margin involvement by pleomorphic lobular carcinoma in situ. AB - CONTEXT: The appropriate treatment for patients with pleomorphic lobular carcinoma in situ (PLCIS) is unknown. When diagnosed on core biopsy, excision is recommended; however, management of PLCIS when it involves margins has not been addressed. OBJECTIVE: To evaluate the significance of PLCIS that is located close to, or at, a resection margin. DESIGN: We identified 26 patients with resection specimens containing PLCIS, all of whom were offered chemoprevention and radiation therapy. The margin status in these patients was subdivided as PLCIS cells at the margin without obvious truncation of lesion; PLCIS less than or equal to 1 mm from, but not involving, the margin; PLCIS 1.1 to 2 mm from the margin; and PLCIS at least 2.1 mm from the margin. RESULTS: Patient age ranged from 35 to 76 years (mean, 58 years), and length of follow-up ranged from 4 to 108 months (mean, 46 months). Six of the 26 patients (23%) received chemoprevention, 4 of 26 patients (15%) received radiation therapy, and 6 of 26 patients (23%) received both. The remaining 10 patients received no further therapy. Pleomorphic lobular carcinoma in situ was at the margin in 6 of the 26 cases (23%), 1 mm from the margin in 7 of 26 cases (27%), 1.1 to 2 mm from the margin in 4 of 26 cases (15%), and was at least 2.1 mm from the margin in 9 of 26 cases (35%). In 1 of the 26 patients, recurrent PLCIS was identified 18 months after initial surgery, for an overall recurrence rate of 3.8%. All other patients were clinically and radiologically free of disease at last follow-up. CONCLUSIONS: This is the first series, to our knowledge, that evaluates margin status in patients with PLCIS and documents recurrence. Recurrent PLCIS was identified at a rate similar to low- or intermediate-grade ductal carcinoma in situ. Therefore, known methods of local control, including surgical excision with negative margins (2 mm), may be the appropriate treatment in these patients. PMID- 21631267 TI - Comparison of formalin-free tissue fixatives: a proteomic study testing their application for routine pathology and research. AB - CONTEXT: Formalin-fixed, paraffin-embedded tissue is the routine processing method for diagnostics practiced in pathology departments worldwide. OBJECTIVE: To determine the potential value of non-cross-linking, formalin-free tissue fixation for diagnostics in pathology and proteomic investigations. DESIGN: We tested 3 commercially available, formalin-free tissue fixatives-FineFIX, RCL2, and HOPE-in lung cancer specimens from 10 patients. The fixatives were evaluated for their effects on tissue morphology, protein recovery, and immunoreactivity for a selected panel of proteins differently expressed in lung cancer, using immunohistochemistry and Western blotting. RESULTS: Tumor-cell analysis with hematoxylin-eosin worked equally well for all tested fixatives when compared with the standard formalin-fixed, paraffin-embedded procedure. Movat pentachrome stains showed comparable results for the different matrices and cellular proteins analyzed. The RCL2 (P = .01) and HOPE fixatives (P = .03) improved protein recovery when compared with formalin-fixed, paraffin-embedded or frozen tissues. Our data clearly show that the fixatives evaluated influenced immunoreactivity to matched, formalin-fixed, paraffin-embedded lung cancer tissue. In particular, membrane-bound proteins, such as epidermal growth factor receptor EGFR, can be detected more efficiently by immunohistochemistry and Western blotting. CONCLUSION: We have demonstrated that formalin-free fixatives have the potential in routine pathology and research to replace formalin in histomorphology and protein preservation. PMID- 21631268 TI - Characterization of CXCR4 expression in chondrosarcoma of bone. AB - CONTEXT: Alterations in molecular elements derived from the CXC chemokine receptor 4 (CXCR4)/stromal-derived factor 1 (SDF-1) cytokine system have been found to strongly correlate with neoplastic progression leading to metastasis in a number of tumors, including osteosarcoma. Excluding hematologic malignancies, chondrosarcoma of bone is the most common primary malignant tumor of bone in adults in the United States. Like osteosarcoma, chondrosarcoma preferentially metastasizes to lung, bone, and very rarely to regional lymph nodes. However, the role of the signal pathway(s) driving neoplastic progression in chondrosarcoma has not yet been clearly elucidated. OBJECTIVE: To test whether CXCR4 was detectable in chondrosarcoma and whether CXCR4 expression levels correlated with chondrosarcoma grade. DESIGN: Twenty-two chondrosarcoma samples banked at our institution between 2001 and 2006 were retrieved for study. By using invasive ductal carcinoma of the breast and osteosarcoma as the positive controls, immunohistochemistry was performed on paraffin-embedded tissue sections and the intensity of the tumor cells was analyzed by morphometric techniques. RESULTS: All chondrosarcoma cases (22 of 22) were immunoreactive for CXCR4. However, the staining intensity of the CXCR4 between the low- and high-grade groups was significantly different. There was a higher staining intensity in high-grade chondrosarcoma cells (P < .001). CONCLUSION: CXCR4 is expressed in chondrosarcomas. CXCR4 expression levels were higher in high-grade chondrosarcoma cells than in low-grade specimens. A larger number of cases will be required to confirm these results and expand the observation, but preliminary data would argue for CXCR4 immunohistochemistry as a potential marker for biologic aggressiveness in chondrosarcoma of bone. PMID- 21631269 TI - Aberrant CpG island hypermethylation in pediatric gastric mucosa in association with Helicobacter pylori infection. AB - CONTEXT: Helicobacter pylori infection is primarily acquired during childhood and persists throughout life in the absence of eradication with antibiotics. Helicobacter pylori infection induces methylation in the promoter CpG island loci in gastric epithelial cells. Thus, aberrant CpG island hypermethylation in gastric epithelial cells likely occurs early in life, although there are no existing data supporting this notion. OBJECTIVES: To identify whether aberrant CpG island hypermethylation occurs in pediatric stomach mucosa in association with H pylori infection and to compare methylation profiles of samples from pediatric and adult stomach tissues. DESIGN: We analyzed pediatric (n = 47) and adult (n = 38) gastric mucosa samples for their methylation status in 12 promoter CpG island loci using the MethyLight assay and compared the number of methylated genes and the methylation levels in individual genes between H pylori -positive and H pylori -negative sample results and between pediatric and adult samples. RESULTS: The average number of methylated genes was significantly higher in H pylori -infected pediatric samples than in H pylori -negative pediatric samples (3.4 versus 0.3, P < .001) and in H pylori -infected adult samples than in H pylori -negative adult samples (7.6 versus 0.9, P < .001). Seven genes showed significantly higher methylation levels in H pylori -infected pediatric samples than in H pylori -negative pediatric samples (all values were P < .05). CONCLUSIONS: These results indicate that CpG island hypermethylation occurs in pediatric gastric mucosa in association with H pylori infection and that the genes affected by H pylori -associated hypermethylation were similar in pediatric and adult samples. PMID- 21631270 TI - Acidophilic nuclear inclusions are specific for florid ductal hyperplasia among proliferative breast lesions. AB - CONTEXT: Recently we have observed distinctive acidophilic intranuclear inclusions in cases of usual intraductal hyperplasia of the breast. Similar inclusions were described almost 20 years ago in cases of mammary hyperplasia. These correlated ultrastructurally with so-called helioid inclusions. However, there since has been little discussion of these inclusions in the literature. OBJECTIVE: To examine the incidence and specificity of these inclusions in proliferative lesions of the breast. DESIGN: Forty cases of usual intraductal hyperplasia, 15 cases of atypical ductal hyperplasia, and 34 cases of low-grade ductal carcinoma in situ were examined for the presence of acidophilic intranuclear inclusions. RESULTS: Acidophilic intranuclear inclusions were present in 50% of cases of usual intraductal hyperplasia (20 of 40) but were not identified in any cases of atypical ductal hyperplasia (0 of 15) or low-grade ductal carcinoma in situ (0 of 34). CONCLUSIONS: Acidophilic intranuclear inclusions appear to be a common, specific feature found in usual intraductal hyperplasia and may be helpful in distinguishing it from atypical ductal hyperplasia and low-grade ductal carcinoma in situ in some cases. Elucidating the nature of these inclusions may provide insight into the pathogenesis of usual intraductal hyperplasia. PMID- 21631271 TI - Uniform expression of Notch1, suppressor of B-cell-specific gene expression, in plasmablastic lymphoma. AB - CONTEXT: Although the loss of B-lineage-specific gene expression is a distinctive feature of plasmablastic lymphoma, the underlying mechanism remains poorly understood. A candidate for this mechanism is Notch1 signaling, which interferes with the activity of B-cell-specific transcription factors E2A and early B-cell factor and positively regulates the mammalian target of rapamycin (mTOR) pathway. OBJECTIVE: To explore the mechanism of loss of B-cell phenotype by correlating expression of B-cell markers with that of Notch1 and downstream targets of the mTOR pathway in plasmablastic lymphoma. DESIGN: A combination of flow cytometric and immunohistochemical immunophenotyping techniques was used on 9 cases of plasmablastic lymphoma to correlate loss of B-cell markers with expression of Notch1 and downstream activation of the mTOR pathway. These results are compared with 5 cases of primary effusion lymphoma and 21 cases of plasma cell myeloma. RESULTS: Plasmablastic lymphoma cases exhibit nearly complete loss of B-cell associated markers and uniform expression of Notch1, with a predominantly nuclear staining pattern. There is a concurrent activation of the mTOR pathway, indicated by expression of mTOR targets eukaryotic initiation factor 4E-binding protein 1 and phosphorylated ribosomal protein S6 in most cases. Similar results are seen in cases of primary effusion lymphoma and plasma cell myeloma. CONCLUSIONS: These findings suggest that activation of Notch1 may be involved in suppression of B cell-specific gene expression and global loss of the B-cell phenotype in plasmablastic lymphoma, similar to primary effusion lymphoma and plasma cell myeloma. Thus, there might be a role for the Notch1 and mTOR pathways in the pathogenesis and therapy of plasmablastic lymphoma. PMID- 21631272 TI - p63 expression in giant cell-containing lesions of bone and soft tissue. AB - CONTEXT: Previous studies have demonstrated p63 overexpression in giant cell tumors of bone and advocate its use as a potential diagnostic marker. Although routine histology is often all that is required to diagnose giant cell tumor of bone, immunohistochemistry could prove useful to distinguish it from other benign and malignant giant cell-containing lesions of bone and soft tissue on needle biopsies and unusual clinical settings. OBJECTIVE: To assess p63 expression in giant cell-containing lesions of bone and soft tissue. DESIGN: p63 immunohistochemistry was performed in 23 giant cell tumors of bone, 8 primary aneurysmal bone cysts, 12 chondroblastomas, 4 giant cell reparative granulomas, 4 osteosarcomas, 15 tenosynovial giant cell tumors, 6 nonossifying fibromas, and 4 pigmented villonodular synovitides. RESULTS: p63 overexpression was identified in 20 of 23 giant cell tumors of bone (86.9%), 5 of 8 primary aneurysmal bone cysts (62.5%), 10 of 12 chondroblastomas (83.3%), 4 of 4 giant cell reparative granulomas (100%), 2 of 4 osteosarcomas (50%), 1 of 15 tenosynovial giant cell tumors (6.6%), 1 of 6 nonossifying fibromas (16.6%), and 1 of 4 pigmented villonodular synovitides (25%). The sensitivity, specificity, positive predictive value, and negative predictive value of p63 immunohistochemistry for the diagnosis of giant cell tumor of bone were 86.95%, 53.36%, 45.45%, and 91.17%, respectively. CONCLUSIONS: This study shows that although p63 is expressed by most giant cell tumors of bone, its lack of specificity limits its use as an immunohistochemical marker in the differential diagnosis of giant cell-containing lesions of bone and soft tissue. PMID- 21631273 TI - Modern concepts on the role of inflammation in pulmonary fibrosis. AB - CONTEXT: Idiopathic pulmonary fibrosis is a uniformly lethal disease with limited biomarkers and no proven therapeutic intervention short of lung transplantation. Pulmonary fibrosis at one time was thought to be a result of inflammation in the lung. Although some forms of pulmonary fibrosis may result from inflammation, idiopathic pulmonary fibrosis is currently thought to result from cell death primarily and inflammation secondarily. OBJECTIVE: To determine the role of inflammation in pulmonary fibrosis in light of our laboratory's published and unpublished research and published literature. DATA SOURCES: Review based on our laboratory's published and unpublished experimental data with relevant background and clinical context provided. CONCLUSIONS: Although cell death is central to pulmonary fibrosis, the proper cytokine environment leading to macrophage polarization is also critical. Evaluation of this environment is promising both for the development of disease biomarkers and for targets for therapeutic intervention. PMID- 21631274 TI - Practical neuropathology synoptic reporting for central nervous system tumors. AB - CONTEXT: Synoptic reporting for central nervous system (CNS) tumors has never been formally addressed, and neuropathologists lack practical templates that they can adapt to their laboratory information system to be compliant with College of American Pathologists (CAP) standards. OBJECTIVES: To provide practical synoptic report templates designed for CNS tumors that allow for easy data extraction and CAP compliance and improve the reporting of CNS tumors. DATA SOURCES: Review of literature and synoptic report format experience in our practice. CONCLUSIONS: Synoptic reporting of required elements is a recently introduced standard for CNS tumors. It is difficult to use a universal non-CNS tumor synoptic report template for CNS tumors because they are heavily weighted to include items not important or required for CNS tumors, such as margins and the TNM classification system. In addition, the CAP CNS protocol, published in 2008, is an immense comprehensive document that is not conducive to simple inclusion in a narrative report. We describe our experience using a synoptic template for CNS tumors that includes all required elements, is tailored to the practice of neuropathology, and can easily be adapted to other laboratory information systems. Because of the multidisciplinary nature of CNS tumor diagnoses, neuropathologists typically collect clinical, demographic, and imaging data on all CNS tumor cases. These data can readily be entered into a primary synoptic report that could replace our standard narrative report. PMID- 21631275 TI - A fatal central nervous system enterovirus 68 infection. AB - The anticipated eradication of poliovirus emphasizes the need to identify other enteroviral causes of severe central nervous system disease. Enterovirus 68 has been implicated only in cases of respiratory illness. We therefore report a case of fatal meningomyeloencephalitis caused by enterovirus 68 in a 5-year-old boy, which required neuropathology, microbiology, and molecular techniques to diagnose. PMID- 21631276 TI - A case of black esophagus with histopathologic description and characterization. AB - Black esophagus, or acute necrotizing esophagitis, is a blackening of the esophagus that is usually distal with a sharp demarcation at the gastroesophageal border. Black esophagus is known to the gastroenterology community; however, to our knowledge it is virtually unknown in the pathology literature with only a single instance described in 1967. It is thought to occur as a poorly elucidated ischemic phenomenon. We report a case of black esophagus in a 45-year-old woman with a history of cocaine and alcohol abuse who was found unresponsive after a vague 2-day illness. On autopsy examination, the esophagus was black with ischemic necrosis of the mucosa, submucosa, and muscularis propria including a diffuse acute inflammatory infiltrate and brown pigmentation limited to the mucosa. Positive periodic acid-Schiff and negative iron stains suggest that the pigment is lipofuscin, likely secondary to ischemia. PMID- 21631277 TI - Blue nevus of the prostate. AB - Blue nevus is one of the melanotic lesions that can incidentally arise in the prostate gland. A literature review identified 28 previously reported cases, and although rare, the blue nevus appeared to be the commonest melanocytic lesion arising in the prostate. The differential diagnosis includes melanosis and malignant melanoma, as well as nonmelanotic lesions due to deposition of lipofuscin, hemosiderin and, rarely, homogentisic acid. The distinction among these lesions can typically be made based on morphologic grounds but may also be aided by histochemical and immunohistochemical stains such as stains for iron, S100 protein, HMB-45, and CD68 as needed. Blue nevus of the prostate is a benign lesion with no malignant potential to date, so no further treatment is warranted. PMID- 21631278 TI - Collagenous sprue: a rare, severe small-bowel malabsorptive disorder. AB - Collagenous sprue is a severe malabsorptive disorder, histologically characterized by small intestinal villous and crypt atrophy, and a subepithelial collagen deposit, thicker than 12 um, that entraps lamina propria cellular elements. Collagenous sprue is a rare disease entity, with only about 60 sporadic cases reported worldwide since it was first described in 1947. Its exact etiology is still under investigation, and its relationship with classic celiac disease and other refractory, spruelike intestinal disorders remains controversial. Two larger-scale studies, in 2009, brought new insights into this elusive, yet emerging, topic. Here, we present a review of the literature on the possible etiology of collagenous sprue, its proposed links to classic celiac disease and to refractory sprue, and its clinical, biochemical, histologic, and molecular features. To our knowledge, all case reports on collagenous sprue in the medical literature to date are summarized. PMID- 21631279 TI - An overview of foot infections in diabetes. AB - Patients with diabetes are prone to ulcerations of the lower extremities, frequently complicated by infection, and are then reliant upon their caregivers for preservation of their limbs without the dreaded outcome of amputation. The enormous tolls of foot infections in diabetes, in terms of both health-related quality of life issues and associated economic burdens, have only been fully realized within the last few decades, and it is anticipated that these burdens will only increase over time. Early and appropriate antibiotic treatment targeting the most likely etiologic pathogens is a cornerstone of management of foot infections in diabetes, but these decisions are now complicated by the emergence of resistant organisms, particularly methicillin-resistant Staphylococcus aureus and multidrug-resistant Gram-negative species. This review will examine the impact of foot infections in diabetes and the overall care and management of the diabetes patient with foot infection, including the potential value of emerging antibiotic therapies within the milieu of antibiotic resistance. PMID- 21631280 TI - Characterization of genetic variation in the Forkhead box class O3 gene (FOXO3) in sheep. AB - FOXO3 is a member of the Forkhead box class O (FOXO) transcription factor family that modulates the transcription of target genes in a variety of cellular functions in lower animals to mammals. It has been implicated in the attenuation of insulin-signaling pathways, including having a potential role in lifespan control in rodents and invertebrates. Recent studies also provide some evidence for an association between ageing, human longevity, and variation in the FOXO3 gene. To date there has been no study on variation in ovine FOXO3 or its relationship to longevity in sheep. In this study, we investigated the variation in exon 2 of ovine FOXO3. This exon encompasses the DNA-binding domain, which is the key regulator of transcription activity in the target genes. To enable investigation of the entire exon 2 of the gene, four overlapping fragments were amplified and analyzed by polymerase chain reaction-single-strand conformational polymorphism and DNA sequencing. Seven haplotypes with 10 nucleotide substitutions were identified across the 1422-bp exon 2 fragment. One of these substitutions was predicted to produce an amino acid change (p.M201V). The variation identified in ovine FOXO3 may have an impact on both the structure and function of ovine FOXO3 and, consequently, affect the longevity of sheep. PMID- 21631281 TI - Comparison of mRNA expression patterns of class B scavenger receptors in BV2 microglia upon exposure to amyloidogenic fragments of beta-amyloid and prion proteins. AB - The inflammatory responses in Alzheimer's disease and prion diseases are dominated by microglia activation. Scavenger receptors have been recently related to the innate immune activation of microglia initiated by endogenous ligands. In this study, we investigated mRNA expression patterns of B class scavenger receptors CD36 and scavenger receptor B1 (SR-B1) in BV2 microglia upon exposure to amyloid fibril Abeta(1-42) and PrP(106-126), respectively. CD36 and SR-B1 showed similar mRNA expression patterns following each treatment. PrP(106-126) induced a rapid increase of CD36 and SR-B1 mRNA levels in the treated microglia, whereas Abeta(1-42) induced a delayed but persistent increase in the mRNA expression of CD36 and SR-B1. These results suggest a possible involvement of CD36 and SR-B1 in microglial interaction with amyloidogenic fragments of beta amyloid and prion proteins. PMID- 21631282 TI - Transcription factor Sp1 is essential for the regulation of the porcine caveolin 1 gene. AB - Caveolin-1 (CAV-1) is a key structural component of caveolae that regulates cholesterol. Employing transgenic techniques to regulate the cholesterol content of pork through CAV-1 is hindered by our lack of knowledge about its regulation. To investigate the regulatory mechanism of porcine CAV-1, a DNA segment containing the 5'-flanking region of CAV-1 was isolated from porcine genomic DNA and sequenced. The luciferase reporter assay detected five cis-acting elements for efficient expression of the CAV-1 gene at the region spanning nucleotides 213 to -20 with serially deleted 5'-flanking sequences and site-directed mutants, -123 to -114 was the core promoter. The electrophoretic mobility shift assay demonstrated potential binding of Sp1 protein to this core promoter. The purpose of this study is to systematically elucidate the transcriptional regulation mechanism of porcine CAV-1 and to contribute to the investigation of the interaction between CAV-1 and cholesterol. PMID- 21631283 TI - H2AFX polymorphisms are associated with decreased risk of diffuse large B cell lymphoma in Koreans. AB - Polymorphisms of the H2A histone family member X (H2AFX) gene have been associated with decreased non-Hodgkin lymphoma (NHL, -417AA) risk and increased breast cancer (1654AG/GG, and -1420GA/AA) risk. We investigated whether H2AFX polymorphisms are associated with the risk of NHL and its subtypes in 573 NHL Korean patients and 721 cancer-free control subjects, using high resolution melting polymerase chain reaction and an automatic sequencer. There was no association between polymorphisms and the risk of overall NHL, all B cell lymphoma, or all T cell lymphoma. However, the -1420 AA genotype was associated with decreased diffuse large B cell lymphoma (DLBCL) risk (OR, 0.65; 95% CI, 0.43 0.97), and there was a trend for allele dose-effect (p-trend=0.03). The -1187 CC genotype was associated with decreased DLBCL risk with borderline significance (OR, 0.70; 95% CI, 0.48-1.02). There was a trend for an allele dose-effect with borderline significance (p-trend=0.06). These results suggest that the -1420 AA genotype of H2AFX may be associated with reduced DLBCL risks in the Korean population. PMID- 21631284 TI - Evaluating a set of reference genes for expression normalization in multiple tissues and skeletal muscle at different development stages in pigs using quantitative real-time polymerase chain reaction. AB - Gene expression analysis requires the use of reference genes consistently expressed under various conditions. In many cases, however, the commonly used reference genes are not uniformly expressed independently of tissues or environmental conditions. To provide a set of reliable reference genes in pigs, we used quantitative polymerase chain reaction to examine expression of six common reference genes (GAPDH, ACTB, H3F3A, HPRT1, RPL32, and RPS18) in adult tissues and prenatal skeletal muscles at 33, 65, and 90 days postcopulation from Tongcheng (obese-type) and Landrace (lean-type) pigs. The expression stability of these reference genes was evaluated by NormFinder, BestKeeper, and geNorm methods. Our data suggest that the reference genes were expressed variably in different tissues, developmental stages and breeds. RPS18, PRL32, and H3F3A could be used as internal controls to normalize gene expression in pig tissues and developmental skeletal muscle. The combination of internal control genes was necessary for accurate expression normalization. During skeletal muscle development, H3F3A and RPS18 would be the most appropriate combination to normalize gene expression in Tongcheng pigs, whereas the combination of PRL32 and RPS18 would be more suitable in Landrace pigs. In different tissues, the expression of PRL32 and RPS18 was the most consistent, and the combination of three genes (RPL32, RPS18, and H3F3A) is the most suitable for accurate normalization. PMID- 21631285 TI - Space radiation damage in ZnO induced by subthreshold electrons: defect identity and optical degradation. AB - Space radiation damage in ZnO induced by subthreshold electrons was studied through reflectance spectra, electron paramagnetic resonance, and photoluminescence. Owing to the vacuum freezing treatment, perturbed singly ionized zinc vacancies (V'(Zn)-), chemisorbed species, and electrons in the conduction band and/or bound to shallow donor levels were observed. V'(Zn)- is due to the ionization and the ionization-induced diffusion processes and is most likely responsible for the 420-nm absorption band. These results also support that the green luminescence in ZnO is related to zinc vacancies. PMID- 21631286 TI - Radiation-induced bystander effects: what are they, and how relevant are they to human radiation exposures? AB - The term radiation-induced bystander effect is used to describe radiation-induced biological changes that manifest in unirradiated cells remaining within an irradiated cell population. Despite their failure to fit into the framework of classical radiobiology, radiation-induced bystander effects have entered the mainstream and have become established in the radiobiology vocabulary as a bona fide radiation response. However, there is still no consensus on a precise definition of radiation-induced bystander effects, which currently encompasses a number of distinct signal-mediated effects. These effects are classified here into three classes: bystander effects, abscopal effects and cohort effects. In this review, the data have been evaluated to define, where possible, various features specific to radiation-induced bystander effects, including their timing, range, potency and dependence on dose, dose rate, radiation quality and cell type. The weight of evidence supporting these defining features is discussed in the context of bystander experimental systems that closely replicate realistic human exposure scenarios. Whether the manifestation of bystander effects in vivo is intrinsically limited to particular radiation exposure scenarios is considered. The conditions under which radiation-induced bystander effects are induced in vivo will ultimately determine their impact on radiation-induced carcinogenic risk. PMID- 21631287 TI - Influence of dental restorative materials on ESR biodosimetry in tooth enamel. AB - Using an experimental model and PENELOPE Monte Carlo simulations, the effects of resin and amalgam on the absorbed doses in tooth enamel were studied to evaluate the feasibility of using restored teeth in electron spin resonance (ESR) dose reconstruction. The model consisted of a phantom containing a plate of these restorative materials placed between powered enamel layers exposed to X rays and a 60Co beam. The experimental results and simulations agreed, showing that the attenuation produced by amalgam and resin with a thickness of 1, 2, and 4 mm is similar to that produced by the enamel itself in the case of the radiation sources employed. For X rays and 60Co gamma radiation the attenuation reached almost 100% and 40%, respectively. These results show that for ESR dose reconstruction, the use of all available enamel of a tooth leads to errors in the estimated dose due to attenuation effects in both healthy and restored teeth. Thus the importance of an enamel selection from different sides of the tooth surface to apply ESR dose reconstruction in the case of a practical situation is shown. PMID- 21631288 TI - Alpha particles induce different F values in monocellular layers of settled and attached human lymphocytes. AB - There is rapidly increasing information on the issue of three-dimensional nuclear architecture, according to which chromosomes are organized in localized territories and chromosome arms in exclusive domains within a given territory. The aim of the present study was to investigate the impact of different cell exposure conditions on cytogenetic damage induced by high-LET radiation. To this end the yield ratio of dicentrics to centric rings (F value) induced by (241)Am alpha particles was analyzed in monolayer cultures of human lymphocytes that were either settled or attached to foils, simulating a rounded or spread out cellular geometry, respectively. Monolayers were exposed in special irradiation chambers to 0.1 and 1.0 Gy and subsequently analyzed for chromosome aberrations. Independent of these different dose levels, significantly different F values of 10.07 +/- 1.73 and 4.27 +/- 0.44 have been determined in attached and settled lymphocytes, respectively. Since the diameter of nuclei vertically traversed by alpha particles in attached cells is about one-half that in settled cells, these F values support the postulate that proximity effects regarding the chromatin geometry in flattened or spherical human lymphocytes influence the formation of high-LET radiation-induced dicentrics and centric rings. A comparison with our earlier data sets obtained for both in vitro and in vivo exposure of human lymphocytes to alpha particles or (137)Cs gamma rays supports the notion that the F value depends on the radiation quality when investigations are confined to spherical human lymphocytes. Thus the F value should not be ruled out as a practical chromosomal "fingerprint" for past exposure to high-LET radiation. PMID- 21631290 TI - Knockdown of cytoglobin expression sensitizes human glioma cells to radiation and oxidative stress. AB - Cytoglobin is a recently identified vertebrate globin whose functions include scavenging reactive oxygen and nitrosative species. In tumor cells, CYGB may function as a tumor suppressor gene. Here we show that knockdown of cytoglobin expression can sensitize human glioma cells to oxidative stress induced by chemical inhibitors of the electron transport chain and as well can increase cellular radiosensitivity. When treated with antimycin A, an inhibitor of the mitochondrial electron transport chain, cytoglobin-deficient cells showed significantly higher H2O2 levels, whereas H2O2 levels were significantly reduced in cytoglobin-overexpressing cells. In addition, cytoglobin knockdown significantly decreased the doubling time of glioma cell lines, consistent with a putative tumor suppressor function. These finding suggest that modulating cytoglobin levels may be a promising treatment strategy for sensitizing human glioma cells to oxidative stress that is induced by ionizing radiation, certain chemotherapies and ischemia-reperfusion. PMID- 21631289 TI - Alpha particles induce apoptosis through the sphingomyelin pathway. AB - The sphingomyelin pathway involves the enzymatic cleavage of sphingomyelin to produce ceramide, a second messenger that serves as a key mediator in the rapid apoptotic response to various cell stressors. Low-linear energy transfer (LET) gamma radiation can initiate this pathway, independent of DNA damage, via the cell membrane. Whether short-ranged, high-LET alpha particles, which are of interest as potent environmental carcinogens, radiotherapies and potential components of dirty bombs, can act through this mechanism to signal apoptosis is unknown. Here we show that irradiation of Jurkat cells with alpha particles emitted by the 225Ac-DOTA-anti-CD3 IgG antibody construct results in dose dependent apoptosis. This apoptosis was significantly reduced by pretreating cells with cholesterol-depleting nystatin, a reagent known to inhibit ceramide signaling by interfering with membrane raft coalescence and ceramide-rich platform generation. The effects of nystatin on alpha-particle-induced apoptosis were related to disruption of the ceramide pathway and not to microdosimetry alterations, because similar results were obtained after external irradiation of the cells with a broad beam of collimated alpha particles using a planar 241Am source. External irradiation allowed for more precise control of the dosimetry and geometry of the irradiation, independent of antibody binding or cell internalization kinetics. Mechanistically consistent with these findings, Jurkat cells rapidly increased membrane concentrations of ceramide after external irradiation with an average of five alpha-particle traversals per cell. These data indicate that alpha particles can activate the sphingomyelin pathway to induce apoptosis. PMID- 21631291 TI - Nitration activates tyrosine toward reaction with the hydrated electron. AB - 3-Nitrotyrosine has been reported as an important biomarker of oxidative stress that may play a role in a variety of diseases. In this work, transient UV-visible absorption spectra and kinetics observed during the reaction of the hydrated electron, e(aq)(-), with 3-nitrotyrosine and derivatives thereof were investigated. The absorption spectra show characteristics of aromatic nitro anion radicals. The absorptivity of radical anion product at 300 nm is estimated to be (1.0 +/- 0.2) * 10(4) M(-1) cm(-1) at pH 7.3. The rate constants determined for the reaction of e(aq)(-) with 3-nitrotyrosine, N-acetyl-3-nitrotyrosine ethyl ester and glycylnitrotyrosylglycine at neutral pH (3.0 +/- 0.3) * 10(10) M(-1) s( 1), (2.9 +/- 0.2) * 10(10) M(-1) s(-1) and (1.9 +/- 0.2) * 10(10) M(-1) s(-1), respectively, approach the diffusion-control limit and are almost two orders of magnitude higher than those for the reactions with tyrosine and tyrosine containing peptides. The magnitude of the rate constants supports reaction of e(aq)(-) at the nitro group, and the product absorbance at 300 nm is consistent with formation of the nitro anion radical. The pH dependence of the second-order rate constant for e(aq)(-) decay (720 nm) in the presence of 3-nitrotyrosine shows a decrease with increasing pH, consistent with unfavorable electrostatic interactions. The pH dependence of the second-order rate constant for formation of radical anion (300 nm) product suggests that deprotonation of the amino group slows the rate, which indicates that deamination to form the 1-carboxy-2-(4 hydroxy-3-nitrophenyl)ethyl radical occurs. We conclude that the presence of the nitro group activates tyrosine and derivatives toward reaction with e(aq)(-) and can affect the redox chemistry of biomolecules exposed to oxidative stress. PMID- 21631293 TI - Bone remodeling surrounding primary teeth in skeletally immature dogs. AB - OBJECTIVE: To quantify remodeling in the bone surrounding fully erupted primary teeth and to compare bone remodeling in the primary and permanent dentitions. MATERIALS AND METHODS: Two bone sections were obtained bilaterally from the maxilla and mandible of the primary molar region of approximately 5-month-old male beagle dogs. Histomorphometric methods were used to estimate the osteonal remodeling in the alveolar and basal regions of the tooth supporting bone. The following variables were calculated: mineral apposition rate (MAR, um/d), mineralizing surface/bone surface (MS/BS, %), bone formation rate (BFR, %/y), and erosion surface/bone surface (ES/BS, %). Comparisons were made between jaws (maxilla vs mandible) and bone types (alveolar vs basal), and data analyzed by analysis of variance (ANOVA) and Tukey-Kramer tests. Remodeling (BFR) surrounding primary teeth was compared to existing data from bone surrounding permanent teeth. RESULTS: The mean and standard deviation BFR values (%/y) were as follows: mandibular alveolar, 44.10 (+/-26.89); maxillary alveolar, 3.54 (+/-3.57); mandibular basal, 22.65 (+/-14.65); and maxillary basal, 12.33 (+/-7.11). The mandibular BFR was significantly (P < .05) higher than the maxillary bone. The BFR of the alveolar bone of primary teeth was not significantly (P = .48) different from the alveolar bone supporting permanent teeth. CONCLUSIONS: The remodeling rate of alveolar bone in skeletally immature dogs was greater in the mandible than in the maxilla and remained unaltered between primary and permanent dentitions. PMID- 21631294 TI - GATA4 specific nonsynonymous single-nucleotide polymorphisms in congenital heart disease patients of Mysore, India. AB - Congenital heart disease (CHD) is the most common type of birth defect, affecting 1% of all live births. The recent exponential increase in the knowledge of medical genetics has revolutionized the understanding of CHDs during the past few decades. GATA4, a transcription factor, is involved in heart development. There are many contradictory reports on involvement of single-nucleotide polymorphisms (SNPs) of GATA4 in the manifestation of CHD. In view of this, an attempt has been made to analyze the known SNPs of GATA4 in Mysore patients with CHD. Of the 308 CHD patients recruited, 100 were screened for SNPs of GATA4 by MassARRAY, which identified 11 SNPs, of which 6 were found in both CHD cases and controls. The other 5 SNPs, c.278G>C (G93A), c.1207C>A (L403M), c.1232C>T (A411V), c.1295T>C (L432S), and c.1180C>G (P394A), were found only in CHD patients. Secondary structure analysis revealed that mutant proteins with the SNPs G93A, L403M, and L432S showed structural changes in their helix, sheet, and turn. Thus, these findings suggest the involvement of specific SNPs of GATA4 in the manifestation of CHD, reported for the first time in an Indian scenario. However, screening for a larger number of CHD patients would help us to establish genotype-phenotype correlation. PMID- 21631295 TI - Frequencies of three CYP2D6 nonfunctional alleles (CYP2D6*3, *4, and *6) within an Iranian population (Mazandaran). AB - Although the frequencies of CYP2D6 nonfunctional alleles have been extensively studied in most populations worldwide, limited information is available for those of the Iranian population. The present study aimed to determine the frequency of three CYP2D6 nonfunctional alleles (CYP2D6*3, *4, and *6) in the Mazandarani ethnic group among the Iranian population. A total of 100 unrelated healthy subjects living in Mazandaran, a Caspian province in the north of Iran, were selected. Lymphocytic genomic DNA was genotyped by a polymerase chain reaction amplification method for detection of three nonfunctional alleles. Finally, the obtained data were used to determine the frequencies of the three alleles, and the results were compared with published data from other populations. The frequencies for CYP2D6 alleles *3, *4, and *6 were 0.5%, 9%, and 0.5%, respectively. Homozygous or compound heterozygous genotypes that predict poor metabolizer phenotype, that is, *4/*4 or *4/*6, were not found in this study. The result of the present study showed that CYP2D6*4 is the major nonfunctional allele found in Mazandarani subjects. In addition, the three inactive alleles of CYP2D6 accounted for 10% of CYP2D6 alleles in our sample versus 0.2%-25.2% reported in other populations. The frequencies of the studied alleles resulted in significant differences between our sample and East Asians, Black-Tanzanians, Saudi Arabians, and Caucasians. PMID- 21631296 TI - A rapid screening method for the identification of a single-nucleotide polymorphism in the carbonic anhydrase VI gene in studies of sensitivity to the bitter taste of 6-n-propylthiouracil. AB - The ability to perceive the bitter taste of 6-n-propylthiouracil (PROP) is a variable phenotype that has been associated with body mass index (in kg/m(2)) and linked to food choice and satiety. PROP-sensitive and -nonsensitive individuals are defined as tasters and nontasters, respectively. Sensitivity to PROP is a heritable trait based on the TAS2R38 gene on chromosome 7q34. In a recent study we demonstrated an association between PROP sensitivity and the single-nucleotide polymorphism (SNP) rs2274333 (+292A/G) within a coding sequence of the gustin/carbonic anhydrase VI gene. The purpose of this study was to develop a rapid and inexpensive screening method for identification of the rs2274333 SNP in individuals with varying sensitivity to PROP. Our results show that the methodology employed allows distinguishing A/G alleles perfectly, with a simple DNA digestion of a polymerase chain reaction fragment covering the SNP site of interest. So, the polymerase chain reaction followed by restriction fragment length polymorphism assay described in this article can be used as an alternative to sequencing in bitter taster status research, and could be employed as a survey tool in nutrigenomic studies. PMID- 21631297 TI - Association between polymorphism of the tumor necrosis factor alpha-308 gene promoter and colon cancer in the Chinese population. AB - Tumor necrosis factor (TNF) is a type of cytokine that inhibits tumorigenesis. Several studies have assessed the relationship between the polymorphism of TNF alpha 308G/A and the susceptibility to colon cancer; however, the results have been controversial. A case-control study was carried out to investigate whether TNF-alpha 308G/A gene polymorphism was associated with the risk of colon cancer in a group of 180 cases and 180 controls from Heilongjiang, China. DNA was isolated from peripheral blood, and gene polymorphisms were analyzed by polymerase chain reaction-restriction fragment length polymorphism. Results of our study showed that colon cancer cases had a significantly higher frequency of TNF-alpha 308AA genotype (odds ratio=9.42, 95% confidence interval=1.18, 75.15; p=0.03) than controls. When stratified by the tumor location, tumor size, growth pattern, differentiation, and stage of colon cancer, no statistically significant results was observed. The present study demonstrated that TNF-alpha 308AA genotype was associated with a higher risk of colon cancer in the Chinese population. Confirmation of these findings in other populations is required. PMID- 21631298 TI - Role of glutathione-S-transferase P1 hypermethylation in molecular detection of prostate cancer. AB - AIM: The aim of this study was to evaluate the diagnostic potential of the quantification of glutathione-S-transferase P1 (GSTP1) gene hypermethylation in molecular detection of prostate cancer in tissue biopsies. METHODS: One hundred fourteen male patients were enrolled in the study; 44 patients with histopathologically confirmed prostate adenocarcinoma, 20 patients with variable degrees of prostate intraepithelial neoplasia, and 50 with benign prostatic hyperplasia, who served as a control group. Real-time quantitative methylation specific polymerase chain reaction was used for assessment of methylation of the promoter region of the GSTP1 gene. RESULTS: Methylation of the GSTP1 promotor was detected in 24% of patients with benign prostatic hyperplasia, 60% of patients with prostate intraepithelial neoplasia, and in 86.3% of prostate adenocarcinoma patients. A statistically significant difference in the GSTP1/MYOD1 (myogenic differentiation 1gene) methylation ratios among the three groups was observed (p=0.0001). At the cutoff value of 9, GSTP1/MYOD1 methylation ratios showed sensitivity in the detection of prostate adenocarcinoma of 71.8% and specificity of 96%. CONCLUSIONS: Methylation of the GSTP1 promotor is a common molecular alteration in prostate cancer that may be a useful adjunct to serum screening tests and digital rectal examination findings and the use of quantitative real time methylation-specific polymerase chain reaction is a promising technique that often distinguishes malignant from nonmalignant prostate disease. PMID- 21631299 TI - Diagnosis of spinal muscular atrophy via high-resolution melting analysis symmetric polymerase chain reaction without probe: a screening evaluation for SMN1 deletions and intragenic mutations. AB - AIM: Spinal muscular atrophy (SMA) is a well-defined autosomal recessive neuromuscular disorder caused by mutations in the survival motor neuron 1 (SMN1) gene. The most frequently observed mutation is a deletion of exon 7, which has been documented in >95% of SMA patients. A novel technique for detecting mutations known as high-resolution melting analysis (HRMA) has rapidly become the tool of choice for screening pathogenic genetic variants. In the present study, we attempt to validate the applicability of HRMA to the detection of exon 7 deletions and other intragenic mutations in SMN1. RESULTS: Three primer sets were adopted in our HRMA screening for deletion of SMN1 exon 7. In screening attempts utilizing two primer sets, the results of HRMA were not compatible with those obtained by polymerase chain reaction-restriction fragment length polymorphism. Therefore, we applied a modified protocol using revised primer sets, which resulted in an absolute compatibility of results between HRMA and polymerase chain reaction-restriction fragment length polymorphism. With regard to screenings for intragenic mutations in SMN1 exon 3, two primer sets were adopted for use in HRMA. In the initial HRMA screening using the first primer set, we failed to identify any intragenic mutations; however, when using a revised primer set, HRMA successfully detected the presence of a c.275G>C mutation. CONCLUSION: HRMA is a simple but versatile tool to add to the existing arsenal of diagnostic techniques that could aid clinicians/researchers in diagnosing SMA. However, as we demonstrate in the present study, the design and selection of primers is of monumental importance in ensuring the successful application of HRMA to screening for pathogenic variants. PMID- 21631300 TI - Associations of the single-nucleotide polymorphisms of the Mina gene with the development of asthma in Chinese Han children: a case-control study. AB - The single-nucleotide polymorphisms (SNPs) of the Mina gene in animals are associated with the development of Th2-mediated diseases. However, there is no information whether the association occurs in humans. This case-control study aimed at examining the potential association of the SNP of the Mina gene with the development of asthma in Chinese Han children. The DNA genotypes and serum immunoglobulin E and interleukin-4 levels of 202 asthmatic patients and 191 nonasthmatic subjects were determined by matrix-assisted laser desorption ionization-time of flight mass spectrometry method and enzyme-linked immunosorbent assay, respectively. We found that the frequency of the T allele of rs4857304, but not rs832081, rs832078, rs9879532, and rs17374916, in the Mina gene in asthmatic patients was significantly higher than that of controls (p = 0.0199). Using a recessive model, we found that the percentage of patients with TT homozygous rs4857304 was significantly higher than that of controls (p = 0.0282, odds ratio=1.568, 95% confidence interval=1.048-2.346). Further, the mean levels of serum immunoglobulin E and interleukin-4 in the patients with TT genotype of rs4857304 were significantly higher than that of patients with the G allele (p = 0.000 and p = 0.03, respectively). Apparently, the T allele of rs4857304 of the Mina gene may be associated with increased risk for the development of asthma in Chinese Han children. PMID- 21631301 TI - Frequency of uridine monophosphate synthase Gly(213)Ala polymorphism in Caucasian gastrointestinal cancer patients and healthy subjects, investigated by means of new, rapid genotyping assays. AB - AIMS: Uridine monophosphate synthase (UMPS) is a fundamental enzyme in pyrimidine synthesis. A single-nucleotide polymorphism, a G-C transversion at the 638th nucleotide, was demonstrated to increase UMPS activity and suggested to have clinical effects. The aims of this study were to set up simple genotyping methods and investigate the UMPS 638G>C polymorphism in the Caucasian population. RESULTS: Two hundred forty-one patients with gastrointestinal cancers and 189 healthy subjects were enrolled. Genomic DNA was extracted from peripheral blood. A polymerase chain reaction-restriction fragment length polymorphism (RFLP) method was implemented using a forward primer incorporating a mismatched base to produce an artificial restriction site and BsrI restriction enzyme digestion; a denaturing high performance liquid chromatography (DHPLC) method was developed to further speed up UMPS genotyping. A 153 bp UMPS gene fragment was successfully amplified and analyzed in all samples. RFLP and DHPLC results showed a 100% match and where confirmed by direct sequencing. UMPS genotype distribution was similar in patients with cancer and control subjects. CONCLUSIONS: Although no association was detected between UMPS variants and gastrointestinal cancer risk in Caucasians, polymerase chain reaction-RFLP with BsrI digestion and DHPLC set up at 59 degrees C are reliable and cost-effective methods to genotype UMPS. PMID- 21631303 TI - A prospective randomized comparison of traditional laparoendoscopic single-site surgery with needlescopic-assisted laparoscopic nephrectomy in the porcine model. AB - BACKGROUND AND PURPOSE: Laparoendoscopic single-site (LESS) surgery produces virtually no scar but is technically challenging because of the loss of triangulation. The objective of this study is to compare classic transumbilical LESS nephrectomy with needlescopic-assisted laparoscopy (NAL) surgery. In doing so, we evaluated whether the addition of a single 2-mm subcostal port could restore triangulation while not jeopardizing recovery or cosmetic outcome in the porcine model. MATERIALS AND METHODS: Ten female farm pigs were randomized to laparoscopic nephrectomy with either LESS or NAL. In LESS, a TriPort was placed through a single 2.5-cm umbilical incision. In NAL, 5- and 10-mm ports were placed in the umbilicus and a 2-mm port was placed in the midclavicular line. Preoperative, perioperative, and postoperative parameters were compared. Variables were analyzed with the Wilcoxon signed-rank test and two-tailed Fisher exact test. Cosmesis was evaluated objectively using the Vancouver Scar Scale and subjectively by a blinded dermatologist. A cost analysis was performed. RESULTS: Estimated blood loss was minimal in both groups (28.8 mL in LESS and 9.4 mL in NAL). Operative time was significantly shorter in NAL (103 vs 150 min; P<0.001). There was no difference in complications (2 vs 1; P=0.500), objective cosmesis (3.9 vs 3.8; P>0.2), or subjective cosmesis (2 vs 3; P=0.500). The NAL protocol had significantly lower disposable equipment costs ($363 vs $1696). CONCLUSIONS: The addition of a 2-mm subcostal port and the restoration of triangulation in the NAL protocol enable shorter operative times, increased surgeon comfort, improved technical ease, and lower costs while maintaining the scarless cosmesis of the traditional LESS protocol. PMID- 21631304 TI - Feasibility and adequacy of robot-assisted lymphadenectomy for renal-cell carcinoma. AB - BACKGROUND AND PURPOSE: The role of lymph node dissection (LND) for renal-cell carcinoma (RCC) is evolving. When clinically negative, nodal disease is rare, but LND remains important in selected patients. Earlier identification of micrometastasis may become beneficial with emerging systemic agents. The ability to perform an adequate LND laparoscopically is uncertain. Open surgical data suggest a minimum of 12 nodes needed to identify most nodal metastases. Robotics may improve adequacy of laparoscopic LND. We report our results with the first reported robot-assisted LND series for RCC. PATIENTS AND METHODS: Robot-assisted LND was performed in 36 patients with RCC by a single surgeon. For right-sided tumors, LND included paracaval, retrocaval, and interaortocaval nodes, and left sided tumors included interaortocaval and periaortic nodes. RESULTS: Mean patient age was 58 years (22-79) with a mean body mass index of 32 kg/m(2) (20-54). Mean tumor size was 7.3 cm with 16 T(3) tumors, including 4 vena caval tumor thrombi. Mean time for LND was 31 minutes, and mean estimated blood loss was 74 mL with no transfusions. Discharge was postoperative day (POD) 1 in 94% and POD 2 in 6%. A mean of 13.9 nodes was obtained with 1 pN+ (2.8%) patient. Mean nodal yield from the first to second half of cases rose from 11 to 16.8 nodes (P=0.02) with 77% having a minimum of 12 nodes in the second half. CONCLUSIONS: Robot-assisted LND for RCC is feasible with adequate nodal yield. Increased yield in later cases may reflect a learning curve. The positivity rate was low as expected, but higher yield was obtained than in the limited laparoscopic literature. PMID- 21631305 TI - Understanding the relationship between language proficiency, language impairment and rehabilitation: Evidence from a case study. AB - The goal of this study was to address the relationship between language proficiency, language impairment and rehabilitation in bilingual Russian-English individuals with aphasia. As a first step, we examined two Russian-English patients' pre-stroke language proficiency using a detailed and comprehensive language use and history questionnaire and evaluated their impairment using the Bilingual Aphasia Test. We then attempted to replicate and extend Kiran and Roberts' study in 2010, examining results of a primarily semantic treatment for anomia in one Russian-English bilingual patient. The patient's ability to name the trained and untrained items in both the trained (English) and untrained (Russian) languages significantly improved by achieving 100% accuracy. Finally, we examined whether improvements observed in treatment were captured by a broader language test such as the Bilingual Aphasia Test. Results are discussed with respect to factors contributing to the successful treatment and the implications of rehabilitation on assessment of language skills as a function of treatment. PMID- 21631306 TI - The effect of literacy on oral language processing: Implications for aphasia tests. AB - Most studies investigating the impact of literacy on oral language processing have shown that literacy provides phonological awareness skills in the processing of oral language. The implications of these results on aphasia tests could be significant and pose questions on the adequacy of such tools for testing non literate individuals. Aiming at examining the impact of literacy on oral language processing and its implication on aphasia tests, this study tested 12 non literate and 12 literate individuals with a modified Amharic version of the Bilingual Aphasia Test (Paradis and Amberber, 1991, Bilingual Aphasia Test. Amharic version. Hillsdale, NJ: Lawrence Erlbaum.). The problems of phonological awareness skills in oral language processing in non-literates are substantiated. In addition, compared with literate participants, non-literate individuals demonstrated difficulties in the word/sentence-picture matching tasks. This study has also revealed that the Amharic version of the Bilingual Aphasia Test may be viable for testing Amharic-speaking non-literate individuals with aphasia when modifications are incorporated. PMID- 21631307 TI - Primary progressive aphasia in a bilingual speaker: a single-case study. AB - We report on the case of an elderly bilingual woman presenting with a diagnosis of primary progressive aphasia. The participant's native language was Friulian (L1), a predominantly oral Romance language, and her second language was Italian (L2), formally learned at primary school in oral and written forms. We investigated her linguistic abilities by means of the Bilingual Aphasia Test ( Paradis, M., & Libben, G. (1987). The assessment of bilingual aphasia. Hillsdale, NJ: Lawrence Erlbaum Associates), which is specifically devised for studying language levels and skills in bilingual/polyglot individuals with aphasia. Specifically, we focused on different tasks extracted from the Bilingual Aphasia Test, targeting phonology, morphology, syntax and lexical semantics. Results show that both languages were affected to a clinically significant degree, but with different profiles in terms of linguistic levels, suggesting the presence of greater phonological, morphological, grammatical and syntactic impairments in L2. Results are discussed in terms of possible dissociations both within the language system of each language and between languages, within the Procedural/Declarative theoretical framework of language acquisition in bilinguals. PMID- 21631308 TI - Effects of cognate status and language of therapy during intensive semantic naming treatment in a case of severe nonfluent bilingual aphasia. AB - As bilingualism becomes less exceptional in the world, and with the growing incidence of stroke and aphasia, a better understanding of how bilingualism affects aphasia recovery is increasingly important. The present study examined the effect of intensive semantic naming therapy in three phases (Spanish, English and mixed) on within- and across-language generalization for cognates and non cognates, in a bilingual individual with chronic, severe expressive aphasia. We hypothesized that cognates would positively influence cross-linguistic generalization, which might be more likely to occur from L2 to L1. Results indicate relative increases in confrontation naming ability in the following conditions: trained versus untrained, L1 versus L2 or mixed and non-cognates versus cognates. This participant demonstrated a pattern of results consistent with a differential recovery pattern in which presentation of treatment in both languages and training of cognates may have promoted interference, thus increasing the activation threshold, and lowering performance under these conditions. PMID- 21631309 TI - Some critical concerns for adapting the Bilingual Aphasia Test to Bahasa Indonesia. AB - One of the most widely spoken languages of the world, Bahasa Indonesia (BI), became standardized as the official language of Indonesia. Based on Malay, it served as lingua franca in various forms throughout the Indonesian archipelago for centuries. Although BI has been habitually learned as a second language, the number of native speakers of BI continues to increase. As a member of the Western Austronesian branch of the Austronesian language family, its grammar and usage bear some resemblance to related languages such as Tagalog. At the same time, certain morphosyntactic and pragmatic characteristics of BI that distinguish it from other languages have been the subject of extensive research and deliberation. For these reasons, the clinical utility of adapting the Bilingual Aphasia Test ( Paradis, M., & Libben, G. (1987) . The assessment of bilingual aphasia. Hillsdale, NJ: Lawrence Erlbaum Associates) is as evident as it is essential. PMID- 21631310 TI - Adapting the Bilingual Aphasia Test to Rarotongan (Cook Islands Maori): Linguistic and clinical considerations. AB - This article describes the adaptation of the Bilingual Aphasia Test (BAT) to the Rarotongan dialect of Cook Islands Maori, a Polynesian language spoken in the Cook Islands and expatriate communities. A brief linguistic sketch of Rarotongan is presented. As Rarotongan is characterised by a complex pronominal system, 'a' versus 'o' possession and optional topicalisation and focus constructions, particular issues arose in obtaining a rigorous adaptation of the BAT. Methods for ensuring effective adaptation across contrastive language pairs and sociocultural aspects of adapting the BAT to Rarotongan are discussed. Obtaining adaptations from several proficient bilingual consultants, comparing versions and group discussion to resolve discrepancies were used for this adaptation and are recommended. It is asserted that every individual has the right to receive accurate, detailed language assessment in each of their languages, irrespective of the languages spoken in the wider community. Further adaptations of the BAT will assist this to be achieved. PMID- 21631311 TI - An automated version of the BAT Syntactic Comprehension task for assessing auditory L2 proficiency in healthy adults. AB - Studies of bilingualism sometimes require healthy subjects to be assessed for proficiency at auditory sentence processing in their second language (L2). The Syntactic Comprehension task of the Bilingual Aphasia Test could satisfy this need. For ease and uniformity of application, we automated its English ( Paradis, M., Libben, G., and Hummel, K. (1987) . The Bilingual Aphasia Test. English version. Hillsdale, NJ: Lawrence Erlbaum Associates) and French ( Paradis, M., & Goldblum, M. C. (1987) . The Bilingual Aphasia Test, French version. Hillsdale, NJ: Lawrence Erlbaum Associates) versions. Although the Bilingual Aphasia Test is meant to assess neurological disorders affecting language, we hypothesised that ceiling performance in L2 would be rare and L2 errors should be consistent with lack of processing automaticity. Initial data from 13 French-English and 4 English-French bilinguals confirm these expectations. Thus, the automated Syntactic Comprehension task (available online for PC and Mac platforms) is indeed suited to test bilingual English and French proficiency levels in healthy adults. PMID- 21631312 TI - Use of the BAT with a Cantonese-Putonghua speaker with aphasia. AB - The aim of this article is to illustrate the use of the Bilingual Aphasia Test (BAT) with a Cantonese-Putonghua speaker. We describe G, who is a relatively young Chinese bilingual speaker with aphasia. G's communication abilities in his L2, Putonghua, were impaired following brain damage. This impairment caused specific difficulties in communication with his wife, a native Putonghua speaker, and was thus a priority for investigation. Given a paucity of standardised tests of aphasia in Putonghua, our goal was to use the BAT to assess G's impairments in his L2. Results showed that G's performance on the BAT subtests measuring word and sentence comprehension and production was impaired. His pattern of performance on the BAT allowed us to generate hypotheses about his higher-level language impairments in Putonghua, which were subsequently found to be impaired. We argue that the BAT is able to capture the primary language impairments in Chinese-speaking patients with aphasia when Putonghua is the second language. We also suggest some modifications to the BAT for testing Chinese-speaking patients with bilingual aphasia. PMID- 21631313 TI - The use of the Bilingual Aphasia Test for assessment and transcranial direct current stimulation to modulate language acquisition in minimally verbal children with autism. AB - Minimally verbal children with autism commonly demonstrate language dysfunction, including immature syntax acquisition. We hypothesised that transcranial direct current stimulation (tDCS) should facilitate language acquisition in a cohort (n = 10) of children with immature syntax. We modified the English version of the Bilingual Aphasia Test (BAT) to test only basic canonical subject-verb-object sentences. We tested syntactic accuracy after teaching then testing all vocabulary from the subsequent syntax test to ensure validity of syntactic scoring. We used scaffolding sentences for syntax training. All procedures were performed both before and after tDCS. Results demonstrated a large effect size of the difference between pre-/post-tDCS groups (p < 0.0005, d = 2.78), indicating syntax acquisition. Combining a modified BAT with tDCS constitutes effective modalities for assessment and treatment of immature syntax in children with autism. Future studies should explore the BAT for patients with an inability to use or understand language, in particular bilingual children with autism. PMID- 21631314 TI - Winning their trust. PMID- 21631315 TI - A model health care delivery system for Medicaid. PMID- 21631317 TI - Ensuring equal access to specialty care. PMID- 21631316 TI - Outcomes of treatment for hepatitis C virus infection by primary care providers. AB - BACKGROUND: The Extension for Community Healthcare Outcomes (ECHO) model was developed to improve access to care for underserved populations with complex health problems such as hepatitis C virus (HCV) infection. With the use of video conferencing technology, the ECHO program trains primary care providers to treat complex diseases. METHODS: We conducted a prospective cohort study comparing treatment for HCV infection at the University of New Mexico (UNM) HCV clinic with treatment by primary care clinicians at 21 ECHO sites in rural areas and prisons in New Mexico. A total of 407 patients with chronic HCV infection who had received no previous treatment for the infection were enrolled. The primary end point was a sustained virologic response. RESULTS: A total of 57.5% of the patients treated at the UNM HCV clinic (84 of 146 patients) and 58.2% of those treated at ECHO sites (152 of 261 patients) had a sustained viral response (difference in rates between sites, 0.7 percentage points; 95% confidence interval, -9.2 to 10.7; P=0.89). Among patients with HCV genotype 1 infection, the rate of sustained viral response was 45.8% (38 of 83 patients) at the UNM HCV clinic and 49.7% (73 of 147 patients) at ECHO sites (P=0.57). Serious adverse events occurred in 13.7% of the patients at the UNM HCV clinic and in 6.9% of the patients at ECHO sites. CONCLUSIONS: The results of this study show that the ECHO model is an effective way to treat HCV infection in underserved communities. Implementation of this model would allow other states and nations to treat a greater number of patients infected with HCV than they are currently able to treat. (Funded by the Agency for Healthcare Research and Quality and others.). PMID- 21631318 TI - Images in clinical medicine. Pulmonary alveolar microlithiasis in a child. PMID- 21631319 TI - Medicine and the epidemic of incarceration in the United States. PMID- 21631320 TI - Integrating neglected tropical diseases into AIDS, tuberculosis, and malaria control. PMID- 21631322 TI - Early-childhood membranous nephropathy due to cationic bovine serum albumin. AB - BACKGROUND: The M-type phospholipase A(2) receptor (PLA(2)R) was recently identified as a candidate antigen in 70% of cases of idiopathic membranous nephropathy, a common form of the nephrotic syndrome. The nature of antigens involved in other idiopathic and secondary membranous nephropathies remains unclear. METHODS: We searched for antibodies against bovine serum albumin and circulating bovine serum albumin by means of enzyme-linked immunosorbent assay and Western blotting in serum specimens obtained from 50 patients with membranous nephropathy and 172 controls. The properties of immunopurified circulating bovine serum albumin obtained from serum specimens were analyzed with the use of two dimensional sodium dodecyl sulfate-polyacrylamide-gel electrophoresis. We detected bovine serum albumin in glomerular deposits and analyzed the reactivity of eluted IgG. RESULTS: Eleven patients, including four children, had high levels of circulating anti-bovine serum albumin antibodies, of both the IgG1 and IgG4 subclasses. These patients also had elevated levels of circulating bovine serum albumin, without an increase in circulating immune complex levels. Bovine serum albumin immunopurified from the serum specimens of these four children migrated in the basic range of pH, whereas the bovine serum albumin from adult patients migrated in neutral regions as native bovine serum albumin. Bovine serum albumin was detected in subepithelial immune deposits only in the children with both high levels of cationic circulating bovine serum albumin and bovine serum albumin specific antibodies, and it colocalized with IgG in the absence of PLA(2)R. IgG eluted from such deposits was specific for bovine serum albumin. CONCLUSIONS: Some patients with childhood membranous nephropathy have both circulating cationic bovine serum albumin and anti-bovine serum albumin antibodies. Bovine serum albumin is present in immune deposits, suggesting that cationic bovine serum albumin is pathogenic through binding to the anionic glomerular capillary wall and in situ formation of immune complexes, as shown in experimental models. PMID- 21631321 TI - Detection of blast-related traumatic brain injury in U.S. military personnel. AB - BACKGROUND: Blast-related traumatic brain injuries have been common in the Iraq and Afghanistan wars, but fundamental questions about the nature of these injuries remain unanswered. METHODS: We tested the hypothesis that blast-related traumatic brain injury causes traumatic axonal injury, using diffusion tensor imaging (DTI), an advanced form of magnetic resonance imaging that is sensitive to axonal injury. The subjects were 63 U.S. military personnel who had a clinical diagnosis of mild, uncomplicated traumatic brain injury. They were evacuated from the field to the Landstuhl Regional Medical Center in Landstuhl, Germany, where they underwent DTI scanning within 90 days after the injury. All the subjects had primary blast exposure plus another, blast-related mechanism of injury (e.g., being struck by a blunt object or injured in a fall or motor vehicle crash). Controls consisted of 21 military personnel who had blast exposure and other injuries but no clinical diagnosis of traumatic brain injury. RESULTS: Abnormalities revealed on DTI were consistent with traumatic axonal injury in many of the subjects with traumatic brain injury. None had detectable intracranial injury on computed tomography. As compared with DTI scans in controls, the scans in the subjects with traumatic brain injury showed marked abnormalities in the middle cerebellar peduncles (P<0.001), in cingulum bundles (P=0.002), and in the right orbitofrontal white matter (P=0.007). In 18 of the 63 subjects with traumatic brain injury, a significantly greater number of abnormalities were found on DTI than would be expected by chance (P<0.001). Follow-up DTI scans in 47 subjects with traumatic brain injury 6 to 12 months after enrollment showed persistent abnormalities that were consistent with evolving injuries. CONCLUSIONS: DTI findings in U.S. military personnel support the hypothesis that blast-related mild traumatic brain injury can involve axonal injury. However, the contribution of primary blast exposure as compared with that of other types of injury could not be determined directly, since none of the subjects with traumatic brain injury had isolated primary blast injury. Furthermore, many of these subjects did not have abnormalities on DTI. Thus, traumatic brain injury remains a clinical diagnosis. (Funded by the Congressionally Directed Medical Research Program and the National Institutes of Health; ClinicalTrials.gov number, NCT00785304.). PMID- 21631323 TI - Saliva polymerase-chain-reaction assay for cytomegalovirus screening in newborns. AB - BACKGROUND: Congenital cytomegalovirus (CMV) infection is an important cause of hearing loss, and most infants at risk for CMV-associated hearing loss are not identified early in life because of failure to test for the infection. The standard assay for newborn CMV screening is rapid culture performed on saliva specimens obtained at birth, but this assay cannot be automated. Two alternatives -real-time polymerase-chain-reaction (PCR)-based testing of a liquid-saliva or dried-saliva specimen obtained at birth--have been developed. METHODS: In our prospective, multicenter screening study of newborns, we compared real-time PCR assays of liquid-saliva and dried-saliva specimens with rapid culture of saliva specimens obtained at birth. RESULTS: A total of 177 of 34,989 infants (0.5%; 95% confidence interval [CI], 0.4 to 0.6) were positive for CMV, according to at least one of the three methods. Of 17,662 newborns screened with the use of the liquid-saliva PCR assay, 17,569 were negative for CMV, and the remaining 85 infants (0.5%; 95% CI, 0.4 to 0.6) had positive results on both culture and PCR assay. The sensitivity and specificity of the liquid-saliva PCR assay were 100% (95% CI, 95.8 to 100) and 99.9% (95% CI, 99.9 to 100), respectively, and the positive and negative predictive values were 91.4% (95% CI, 83.8 to 96.2) and 100% (95% CI, 99.9 to 100), respectively. Of 17,327 newborns screened by means of the dried-saliva PCR assay, 74 were positive for CMV, whereas 76 (0.4%; 95% CI, 0.3 to 0.5) were found to be CMV-positive on rapid culture. Sensitivity and specificity of the dried-saliva PCR assay were 97.4% (95% CI, 90.8 to 99.7) and 99.9% (95% CI, 99.9 to 100), respectively. The positive and negative predictive values were 90.2% (95% CI, 81.7 to 95.7) and 99.9% (95% CI, 99.9 to 100), respectively. CONCLUSIONS: Real-time PCR assays of both liquid- and dried-saliva specimens showed high sensitivity and specificity for detecting CMV infection and should be considered potential screening tools for CMV in newborns. (Funded by the National Institute on Deafness and Other Communication Disorders.). PMID- 21631324 TI - gp100 peptide vaccine and interleukin-2 in patients with advanced melanoma. AB - BACKGROUND: Stimulating an immune response against cancer with the use of vaccines remains a challenge. We hypothesized that combining a melanoma vaccine with interleukin-2, an immune activating agent, could improve outcomes. In a previous phase 2 study, patients with metastatic melanoma receiving high-dose interleukin-2 plus the gp100:209-217(210M) peptide vaccine had a higher rate of response than the rate that is expected among patients who are treated with interleukin-2 alone. METHODS: We conducted a randomized, phase 3 trial involving 185 patients at 21 centers. Eligibility criteria included stage IV or locally advanced stage III cutaneous melanoma, expression of HLA*A0201, an absence of brain metastases, and suitability for high-dose interleukin-2 therapy. Patients were randomly assigned to receive interleukin-2 alone (720,000 IU per kilogram of body weight per dose) or gp100:209-217(210M) plus incomplete Freund's adjuvant (Montanide ISA-51) once per cycle, followed by interleukin-2. The primary end point was clinical response. Secondary end points included toxic effects and progression-free survival. RESULTS: The treatment groups were well balanced with respect to baseline characteristics and received a similar amount of interleukin 2 per cycle. The toxic effects were consistent with those expected with interleukin-2 therapy. The vaccine-interleukin-2 group, as compared with the interleukin-2-only group, had a significant improvement in centrally verified overall clinical response (16% vs. 6%, P=0.03), as well as longer progression free survival (2.2 months; 95% confidence interval [CI], 1.7 to 3.9 vs. 1.6 months; 95% CI, 1.5 to 1.8; P=0.008). The median overall survival was also longer in the vaccine-interleukin-2 group than in the interleukin-2-only group (17.8 months; 95% CI, 11.9 to 25.8 vs. 11.1 months; 95% CI, 8.7 to 16.3; P=0.06). CONCLUSIONS: In patients with advanced melanoma, the response rate was higher and progression-free survival longer with vaccine and interleukin-2 than with interleukin-2 alone. (Funded by the National Cancer Institute and others; ClinicalTrials.gov number, NCT00019682.). PMID- 21631325 TI - Trends in hospital volume and operative mortality for high-risk surgery. AB - BACKGROUND: There were numerous efforts in the United States during the previous decade to concentrate selected surgical procedures in high-volume hospitals. It remains unknown whether referral patterns for high-risk surgery have changed as a result and how operative mortality has been affected. METHODS: We used national Medicare data to study patients undergoing one of eight different cancer and cardiovascular operations from 1999 through 2008. For each procedure, we examined trends in hospital volume and market concentration, defined as the proportion of Medicare patients undergoing surgery in the top decile of hospitals by volume per year. We used regression-based techniques to assess the effects of volume and market concentration on mortality over time, adjusting for case mix. RESULTS: Median hospital volumes of four cancer resections (lung, esophagus, pancreas, and bladder) and of repair of abdominal aortic aneurysm (AAA) rose substantially. Depending on the procedure, higher hospital volumes were attributable to an increasing number of cases nationwide, an increasing market concentration, or both. Hospital volumes rose slightly for aortic-valve replacement but fell for coronary-artery bypass grafting and carotid endarterectomy. Operative mortality declined for all eight procedures, ranging from a relative decline of 8% for carotid endarterectomy (1.3% mortality in 1999 and 1.2% in 2008) to 36% for AAA repair (4.4% in 1999 and 2.8% in 2008). Higher hospital volumes explained a large portion of the decline in mortality for pancreatectomy (67% of the decline), cystectomy (37%), and esophagectomy (32%), but not for the other procedures. CONCLUSIONS: Operative mortality with high-risk surgery fell substantially during the previous decade. Although increased market concentration and hospital volume have contributed to declining mortality with some high-risk cancer operations, declines in mortality with other procedures are largely attributable to other factors. (Funded by the National Institute on Aging.). PMID- 21631326 TI - Intravenous thrombolytic therapy for acute ischemic stroke. PMID- 21631327 TI - Images in clinical medicine. Amyand's hernia. PMID- 21631328 TI - Clinical problem-solving. A recurrent problem. PMID- 21631330 TI - Milk and membranous nephropathy. PMID- 21631329 TI - Brain injuries from blasts. PMID- 21631331 TI - Induced pluripotent stem cells--a cautionary note. PMID- 21631332 TI - HPV vaccine against HPV infection and disease in males. PMID- 21631333 TI - HPV vaccine against HPV infection and disease in males. PMID- 21631334 TI - HPV vaccine against HPV infection and disease in males. PMID- 21631337 TI - Body-mass index and risk of death in Asians. PMID- 21631338 TI - Body-mass index and risk of death in Asians. PMID- 21631340 TI - Cytarabine dose for acute myeloid leukemia. PMID- 21631341 TI - Cytarabine dose for acute myeloid leukemia. PMID- 21631342 TI - Cytarabine dose for acute myeloid leukemia. PMID- 21631344 TI - The ClinicalTrials.gov results database. PMID- 21631345 TI - The ClinicalTrials.gov results database. PMID- 21631346 TI - The ClinicalTrials.gov results database. PMID- 21631347 TI - The ClinicalTrials.gov results database. PMID- 21631349 TI - Intraosseous catheter placement in children. PMID- 21631351 TI - Closed-eye ocular injuries in the Iraq and Afghanistan wars. PMID- 21631354 TI - The role of nuclear factor kappaB in the interferon response. AB - The nuclear factor kappaB (NF-kappaB) transcription factor regulates the expression of genes involved in cell survival and immune responses. We have identified a novel interferon (IFN)-activated signaling pathway that leads to NF kappaB activation and demonstrate that a subset of IFN-stimulated genes and microRNAs that play key roles in cellular response to IFN is regulated by NF kappaB. This review focuses on the IFN-induced NF-kappaB activation pathway and the role of NF-kappaB in the expression of IFN-induced coding and noncoding genes, antiviral activity and apoptosis, and the therapeutic application of IFN in cancer and infectious disease. PMID- 21631355 TI - Macrophages in allergic asthma: fine-tuning their pro- and anti-inflammatory actions for disease resolution. AB - Macrophages exert prominent effects in the defense of the respiratory tract from airborne pathogens. These cells are specialized to recognize, phagocytose, and destroy these infectious agents and then promote appropriate tissue repair after successful pathogen clearance. For reasons that are not presently clear, macrophages appear to be inappropriately activated during asthma responses. Evidence stems from the appearance of either classically (or M1) and alternatively activated (or M2) cells in the alveolar compartment of asthmatic lung. Macrophages localized in the interstitial area of the lung appear to be less prone to polarization toward either the M1 or M2 phenotype as these cells predominately express interleukin-10 and exhibit immunoregulatory properties. Effective treatment of clinical asthma, regardless of severity, might depend on restoring an appropriate balance between M1, M2, and immunoregulatory macrophages in the lung. PMID- 21631356 TI - Laparoscopy rein and a backward needle entrance. AB - Since the awakening of single-port laparoscopy and culdolaparoscopy and with the increased interest in minilaparoscopy-assisted natural orifice surgery and pure natural orifice peritoneoscopy, laparoscopists are in pursuit of techniques with less percutaneous assistance. We made a rein using a 6-cm straight cutting edge needle in a 2-0 nylon suture with a large clip tied near the end as stoppage. We are also presenting an optional technique of placing the 6-cm straight needle backward, when the diameter of the cannula does not allow the parallel passage of the needle holder. The rein is simple, inexpensive, and easy to assemble, and may solve some problems of traction and triangulation without additional ports. PMID- 21631357 TI - Antihypertensive and cardiovascular effects of catechin-rich oil palm (Elaeis guineensis) leaf extract in nitric oxide-deficient rats. AB - Oil palm (Elaeis guineensis) leaf extract (OPLE) possesses good ex vivo vasodilation and antioxidant properties. This study evaluated the catechin-rich OPLE antioxidant, antihypertensive, and cardiovascular effects in normal and nitric oxide (NO)-deficient hypertensive rats. OPLE was administered orally (500 mg/kg of body weight/day) to normotensive Wistar rats and N(omega)-nitro-L arginine methyl ester (L-NAME)-induced NO-deficient hypertensive rats. OPLE significantly (P<.05) attenuated blood pressure increases, increased serum NO, reduced lipid peroxidation, and showed antioxidant effects in NO-deficient hypertensive rats. OPLE decreased the coronary arteriole wall-to-lumen ratio to near normal values under NO deficiency. Although OPLE showed good antihypertensive and antioxidant effects under NO deficiency, it was not hypotensive to normal rats and produced no chronic cardiovascular toxicity in any of the rats throughout the 12-week study. This is the first report on the in vivo antihypertensive properties of green tea catechins extracted from an alternative source, namely, oil palm leaf, for use as a medicinal food for hypertension and cardiovascular ailments. PMID- 21631358 TI - Phenotypic and functional maturation of dendritic cells induced by polysaccharide isolated from Paecilomyces cicadae. AB - Paecilomyces cicadae Miquel Samson is the anamorph of Cordyceps cicadae Shing and is used in functional foods for the prevention and treatment of various diseases. In the present study, we examined the effects of P. cicadae polysaccharide (PCP) on dendritic cell (DC) maturation. Phenotypic maturation of DCs by PCP was confirmed by the elevated expressions of CD80, CD86, major histocompatibility complex (MHC)-I, and MHC-II molecules and functional maturation by increased expression of interleukin-12, interleukin-1beta, and tumor necrosis factor-alpha, enhanced allogenic T cell stimulation, and decreased endocytosis. PCP induced the maturation of DCs from C3H/HeN and C57BL/6 mice but not from Toll-like receptor (tlr) 4-/- knockout mice and TLR4-mutated C3H/HeJ mice, which suggests that TLR4 is the membrane receptor for PCP. PCP increased the degradation of inhibitor of nuclear factor-kappaB (NF-kappaB) alpha/beta, which enhanced the nuclear translocation of NF-kappaB p50/p65 and induced the phosphorylation of extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38 mitogen activated protein kinases, which are signaling molecules downstream of TLR4. These results indicate that PCP induces DC maturation through TLR4 signaling. PMID- 21631359 TI - Punica granatum protects against oxidative stress in PC12 cells and oxidative stress-induced Alzheimer's symptoms in mice. AB - Alzheimer's disease (AD) is a progressive degenerative brain disorder that is characterized by neuronal loss, neurofibrillary tangles, and the abnormal deposition of senile plaque and amyloid beta peptide (Abeta). The brains of AD patients are under intense oxidative stress. The overproduction of Abeta leads to Abeta-associated free radical oxidative stress. In this study, the antioxidative and neuronal protective effects of Punica granatum extract were investigated against oxidative stress-induced cytotoxicity in PC12 cells. The ethanol extracts of P. granatum protected PC12 cells from hydrogen peroxide (H2O2-induced oxidative stress. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction assays revealed a significant increase in cell viability when oxidatively stressed PC12 cells were treated with the P. granatum extract. To examine the effects of P. granatum on Abeta1-42-induced learning and memory impairment in mice, in vivo behavioral tests were performed. Treatment with the extract of P. granatum increased step-through latency in mice injected with Abeta1-42. The results of this study suggest that the ethanol extract of P. granatum mitigated H2O2-induced oxidative stress in PC12 cells. In addition, the extract inhibited neuronal cell death caused by Abeta-induced oxidative stress and Abeta-induced learning and memory deficiency. PMID- 21631360 TI - The effects of powdered fertilized eggs on depression. AB - This 12-week, double-blind, placebo-controlled study investigated the effects of fertilized egg powder (Young Tissue Extract; YT(r)) intake on outcome measures for depression. Fifty-five patients with depression were randomly assigned to receive YTE, YTE plus Melissa officinalis, or placebo for 12 weeks. At baseline, there were no significant differences in scores on the Hamilton Depression Rating Scale (HAM-D) or Beck Depression Inventory II (BDI-II) among the 3 groups. At 12 weeks, the HAM-D scores in groups treated with YTE or YTE with M. officinalis were both significantly lower than those in the placebo group. In addition, both treatment groups showed a significant improvement in depression as measured by the change in HAM-D scores from baseline to 12 weeks, whereas the placebo group showed no significant change. There were no significant differences between the 2 treatment groups. The study indicates that the fertilized egg powder has an antidepressive effect and may be an alternative or adjunct to antidepressive medication for some patients, but further research is necessary. PMID- 21631361 TI - Mulberry leaf extract reduces postprandial hyperglycemia with few side effects by inhibiting alpha-glucosidase in normal rats. AB - Mulberry (Morus alba L.) leaf extract (MLE) was investigated as a potent plant derived alpha-glucosidase inhibitor with low alpha-amylase inhibitory activity. MLE was prepared by heating in an autoclave at 121 degrees C for 15 minutes, and its in vitro and in vivo antihyperglycemic activities were investigated. The adverse side effects of MLE were analyzed by measuring the weight and volume of the cecum, stool color, starch content in the cecum, and the integrity of intestinal transporting capacity. The in vitro inhibitory activity of MLE on intestinal alpha-glucosidase was potent and that on intestinal alpha-amylase was very weak compared with acarbose. Sugar loading tests with starch, maltose, and sucrose showed that MLE may reduce postprandial increases in blood glucose by acting as an intestinal alpha-glucosidase inhibitor. Feeding tests suggested that MLE may exhibit fewer adverse side effects than other alpha-glucosidase inhibitors, such as abdominal flatulence and meteorism, which are attributed to the impaired digestion of starch by strong inhibition of intestinal alpha amylase. These results suggest that MLE could be used in the development of pharmaceutical foods to control the blood glucose levels of diabetic patients by inhibiting intestinal alpha-glucosidase with reduced side effects. PMID- 21631362 TI - Dietary spinach saponin-enriched lipophilic fraction inhibits platelet aggregation and blood coagulation. AB - In this study, we investigated the effect of spinach saponin-enriched lipophilic fraction (SSEF) on collagen (10 MUg/mL)-stimulated platelet aggregation in vivo. Dietary SSEF dose-dependently inhibited collagen-induced platelet aggregation by decreasing thromboxane A2 production and intracellular Ca2+ agonist activity as an aggregation-inducing autacoidal molecule. In addition, SSEF significantly increased the formation of cyclic AMP and cyclic GMP, intracellular Ca2+ antagonists that are aggregation-inhibiting molecules in collagen-stimulated platelets. These results suggest that SSEF is a potent inhibitor of collagen stimulated platelet aggregation in vivo. Prothrombin time and activated partial thromboplastin time, indicators of blood coagulation, were potently prolonged by dietary SSEF in vivo. These findings suggest that SSEF prolongs the interval time between the conversion of fibrinogen to fibrin. Dietary SSEF also inhibited 0.4 M sucrose-induced hemolysis. Accordingly, our data demonstrate that SSEF might be a useful tool for inhibiting platelet activation and blood coagulation in thrombotic diseases. PMID- 21631363 TI - Efficacy of epigallocatechin-3-gallate and Amla (Emblica officinalis) extract for the treatment of diabetic-uremic patients. AB - Uremic patients with diabetes suffer from high levels of oxidative stress due to regular hemodialysis therapy (neutrophil activation induced by hemo incompatibility between the hemodialyser and blood) and complications associated with diabetes. Several plasma biomarkers were screened in 13 uremic diabetic patients after receiving the mixture of (-)-epigallocatechin gallate (EGCG), a major component of green tea extract, and Amla extract (AE), from Emblica officinalis, the Indian gooseberry, for 3 months. We found that oral administration of a 1:1 mixture of EGCG and AE for 3 months significantly improved antioxidant defense as well as diabetic and atherogenic indices in uremic patients with diabetes. Furthermore, no significant changes in hepatic function, renal function, or inflammatory responses were observed. These results suggest that a 1:1 combination of EGCG and AE is a safe and effective treatment for uremic patients with diabetes. PMID- 21631364 TI - Biocompatibility of pooled human immunoglobulin (Gamunex 10%TM) with ocular infusion solutions (BSSTM and BSS PlusTM): an in vitro evaluation of a potential antitoxin treatment for infectious endophthalmitis. AB - PURPOSE: Gamunex 10% (Talecris Biotherapeutics, Research Triangle Park, NC), a commercially available preparation of pooled human immunoglobulin G, has been proposed as an antitoxin therapy against bacterial toxins released in infectious endophthalmitis. Its biocompatibility with two commonly used intraocular infusion fluids was evaluated to determine feasibility of its clinical application in endophthalmitis treatment. METHODS: Gamunex 10% was mixed with BSS or BSS Plus (Alcon Laboratories, Fort Worth, TX) such that it constituted a range of 1.25% 50% by volume. Osmolality, pH, optical density, and ionic strength were measured across this range of concentrations. RESULTS: The amount of pH reduction with increasing concentrations of Gamunex 10% was similar for both BSS and BSS Plus. In BSS Plus, solutions containing up to 20% by volume of Gamunex 10% remained at near-physiologic pH (~7.0 or above). No physiologically significant changes in osmolality or optical density measurements that would be anticipated to have profound physiological effects were observed at any of the measured concentrations, nor was there visual evidence of tubidity/precipitation. A gradual increase in ionic strength was observed with increasing concentrations of Gamunex 10%. CONCLUSIONS: Potentially therapeutic mixtures of Gamunex 10% in 2 commonly used intraocular infusion fluids, BSS and BSS Plus, showed no evidence of bioincompatibility when the solutions were evaluated for changes in osmolality, pH, ionic strength, aggregation, or precipitation. PMID- 21631365 TI - Effect of brimonidine on retinal blood flow autoregulation in primary open-angle glaucoma. AB - PURPOSE: To determine whether topically applied brimonidine affects the retinal hemodynamic autoregulatory response to posture change in patients with normal tension glaucoma. METHODS: Six patients with normal tension glaucoma (primary open-angle glaucoma and maximum untreated intraocular pressure <22 mmHg) in each eye were studied. We retrospectively reviewed retinal hemodynamic data acquired when the patients were off and on treatment with brimonidine 0.15% (twice a day, both eyes) during the course of their care. At each testing session, vessel diameter and blood speed at the same site along the inferior temporal retinal artery of 1 eye were measured while sitting, while reclining for 30 min, and again while sitting using a retinal laser Doppler instrument. Blood flow was computed automatically. Brachial artery blood pressure and heart rate were also measured. The Wilcoxon signed rank test was used to assess the statistical significance of the differences in each measured parameter while subjects were on and off brimonidine. RESULTS: Off brimonidine, the mean blood flow rate increased by 68.0%+/-34.3% (range: +17% to +108%) after 30 min in reclined posture compared to baseline-seated measures. On brimonidine, the mean blood flow rate increased by 8.9%+/-16.8% (range: -9.7 to +28.0%) after 30 min in reclined posture compared to baseline-seated measures. The difference in the posture-induced changes for blood flow rate while on brimonidine compared to off brimonidine was statistically significant (P=0.027). CONCLUSIONS: Off brimonidine, the patients exhibited marked increases in retinal blood flow while reclining. On brimonidine, the hemodynamic changes were consistent with normal autoregulatory control of retinal blood flow. PMID- 21631366 TI - Comparison of intravitreal bevacizumab and ranibizumab treatment for diabetic macular edema. AB - AIM: The aim of this study was to compare the effects of bevacizumab and ranibizumab on visual function and macular thickness in patients with diabetic macular edema (DME). METHODS: The data of diabetic patients who had been treated with bevacizumab for DME were reviewed. Those patients who received 1 injection of intravitreal bevacizumab and ranibizumab with at least 6-month interval were considered for enrollment. The best-corrected visual acuity (BCVA) assesment with Early Treatment Diabetic Retinopathy Study (ETDRS) chart and central subfield macular thickness (CSMT) measurement using optical coherence tomography-3 before and after the injections were recorded as outcome measures. RESULTS: The study included 29 eyes of 29 patients with a mean age of 56.18+/-13.07 years. The median BCVA was 59 ETDRS letters and the median CSMT was 411 MUm preceeding the bevacizumab injection. At the 4th-6th week control after the injection, median BCVA increased to 61.50 ETDRS letters and the median CSMT decreased to 373 MUm. This change in BCVA and CSMT was found to be statistically significant (P=0.029 and P=0.011, respectively). The mean interval between bevacizumab and ranibizumab treatment was 9.54+/-2.64 months. Ranibizumab treatment increased the median BCVA from 53 to 66 ETDRS letters and decreased the median CSMT from 428 MUm to a level of 279 MUm, which were statistically significant (P<0.001 and P<0.001, respectively). The median change in BCVA was 4.5 ETDRS letters in the bevacizumab group and 6 ETDRS letters in the ranibizumab group (P=0.58), whereas the median changes in CSMT were 41 and 100 MUm after bevacizumab and ranibizumab injections, respectively (P=0.005). CONCLUSIONS: Bevacizumab and ranibizumab are both effective antivascular endothelial growth factor drugs preferred in the treatment of DME. Our comparison of both therapies on the same patients suggested that the effect on BCVA was not statistically different, but ranibizumab provided more decrease in CSMT. PMID- 21631367 TI - Patient-clinician communication about end-of-life care topics: is anyone talking to patients with chronic obstructive pulmonary disease? AB - BACKGROUND: Reports describe patient and health care system benefits when clinicians engage in end-of-life conversations with patients diagnosed with life limiting illnesses, yet most clinicians focus on life-preserving treatments and avoid conversations about end-of-life care. We describe patient-clinician communication practices about end-of-life care in patients with chronic obstructive pulmonary disease (COPD) using self-report questionnaires to: (1) characterize the content of patient-clinician communication about end-of-life care from the patient perspective, including topics that were not addressed and ratings of the quality of the communication for topics discussed and (2) determine whether clinician characteristics was associated with the absence of specific communication items addressed. METHODS: Cross-sectional study of outpatients (n = 376) who completed the Quality of Communication (QOC) questionnaire (outcome measure). The primary exposure was clinician training. We used logistic regression. All tests were two-tailed and p < 0.05 was considered significant. RESULTS: Clinicians (n = 92) were staff physicians (33.7%), physician trainees (35.9%), and advanced practice nurses (30.4%). Patients were older (mean age, 69.4 years, standard deviation [SD] 10.0); white (86%) men (97%) with severe COPD (mean forced expiraory volume in 1 second [FEV(1)] percent predicted 50%, SD 20). All end-of-life topics were underaddressed. Four topics were not addressed 77%-94% of the time. None of the QOC items varied significantly by clinician type in adjusted logistic regression. CONCLUSIONS: All end-of-life communication topics were underaddressed by clinicians, regardless of training, with four topics particularly unlikely to be discussed. End-of-life topics that are important to patients should be targeted for an intervention to facilitate improvement in clinicians' communication skills and practice and may improve patient satisfaction with clinician communication. PMID- 21631368 TI - Paroxysmal sympathetic hyperactivity (sympathetic storm) in a patient with permanent vegetative state. AB - BACKGROUND: There are many people living in a permanent vegetative state in the United States. We report the first case of a patient in permanent vegetative state (PVS) who developed paroxysmal sympathetic hyperactivity (sympathetic storm) 8 months after the hypoxic brain injury that lead to PVS. DISCUSSION: Paroxysmal sympathetic hyperactivity is a relatively common complication early in the course of traumatic brain injury. Recognition of the clinical presentation of this syndrome is important to palliative and hospice care providers who may be caring for patients with PVS. The treatment of sympathic hyperactivity to reduce potential physical suffering includes medications targeted to the sympathetic nervous system. PMID- 21631369 TI - Neostigmine for the treatment of gastrointestinal atony: a report of one case. AB - BACKGROUND: A case of severe constipation is described in a 75-year- old cancer patient receiving methadone for pain. Her constipation was refractory to the current treatment and she suffered severe discomfort and cognitive impairment. Due to the severity of the clinical situation and after excluding mechanical obstruction, low doses of subcutaneous neostigmine were administered, having bowel movements with evacuation of stools in a few hours after its administration. DISCUSSION: The results suggest that subcutaneous neostigmine could be an alternative choice in a group of selected patients with advanced cancer and opioid-induced constipation. PMID- 21631371 TI - Efficacy and undesirable effects of corticosteroid therapy experienced by palliative care specialists in Japan: a nationwide survey. AB - BACKGROUND AND METHODS: Corticosteroids are commonly used for symptom relief in the treatment of patients with advanced cancer. Consistent efficacy of corticosteroid treatment in palliative care remains controversial. A cross sectional anonymous survey was mailed to representative managing physicians in certified palliative care units in Japan to clarify the physician-perceived efficacy of steroid treatment on anorexia, fatigue, and dyspnea in terminal cancer patients, to clarify physicians' experience of side effects of corticosteroid use, and to determine the Japanese palliative care physician reported predictive factors for efficacy and lack of efficacy. RESULTS AND CONCLUSIONS: Many Japanese palliative care specialists perceived that corticosteroids are effective for each of the symptoms, are aware of the prevalence and importance of serious adverse effects, and predict the effectiveness of steroid therapy by etiological factors. PMID- 21631370 TI - Parental bereavement needs in the pediatric intensive care unit: review of available measures. AB - BACKGROUND: Pediatric intensive care units (PICUs) are highly technological settings in which advanced care is used to restore health to critically ill children; however, they are also places where children die. Understanding the needs of parents bereaved in this setting is essential for better family care. OBJECTIVE: To systematically review the extant literature to identify instruments potentially useful for assessing the needs of parents bereaved in the PICU. METHODS: We searched PubMedTM, CINAHLTM, and Health and Psychosocial InstrumentsTM for tools to assess family needs during a relative's hospitalization. From 357 abstracts, 96 articles were reviewed that described 31 instruments. Fifteen instruments were selected based on their (1) use with parents and/or the bereaved, (2) use in PICU, neonatal intensive care, or pediatric wards, (3) measurement of family needs or related constructs, and (4) published psychometrics. Need-related constructs included satisfaction with family care and environmental stress since these have been related to met and unmet needs, respectively. RESULTS: No instruments specifically designed to assess the needs of parents bereaved in the PICU were identified. Most tools reviewed showed validity and reliability in the populations and settings for which the tools were intended; however, validity and reliability were not established for parents bereaved in the PICU. No tools addressed the full range of needs for parents bereaved in the PICU. CONCLUSIONS: A new instrument is needed to adequately assess the needs of parents bereaved in the PICU. Patient conditions, illness trajectories, and life course perspectives must be considered in designing a new tool. PMID- 21631372 TI - Tailoring combination oral contraceptives to the individual woman. AB - BACKGROUND: Over the last 50 years, there has been intense interest in the type of progestin used in combined oral contraceptives (COC) in an attempt to exploit novel properties and minimize adverse events. At the same time, the dose of synthetic estrogen, ethinylestradiol, in COC pills has been reduced to minimize risks for ischemic stroke, myocardial infarction, and venous thromboembolism. New formulations, including extended-cycle or continuous-use COCs or those that use a natural estrogen, estradiol, may offer improvements over their predecessors. METHODS: A Medline search was performed to encompass studies published since 1990 that pertain to the pharmacology of estrogens and progestins used in COCs, risks and adverse events associated with COCs, and extended or continuous-use COCs. RESULTS: New progestins structurally related to progesterone and spironolactone may exhibit more selective binding to the progesterone receptor and lack androgenic adverse effects associated with progestins structurally related to testosterone. Recently, COCs containing natural (17beta-estradiol) or conjugated estrogen (estradiol valerate) rather than ethinylestradiol have been developed in order to move to a more natural estrogen. Although many of the new progestins incorporated into COCs have not demonstrated the negative effects on lipid metabolism and other adverse events associated with the traditional progestins, the goal of attaining good cycle control has yet to be achieved. Extended-cycle and continuous-use regimens are now available that reduce the frequency of menses, but breakthrough bleeding remains a problem. CONCLUSIONS: Appropriate counseling to raise awareness of the specific characteristics of the different COC options available may optimize adherence and patient acceptability. PMID- 21631373 TI - Is Spanish language a barrier to domestic violence assessment? AB - BACKGROUND: Domestic violence is the leading cause of injury to premenopausal women and fatal in over 1000 women annually, but few healthcare providers ask about it, citing numerous barriers, including language. This study tested the hypothesis that language does, in fact, pose a barrier to screening and that Spanish-speaking women report lower lifetime screening rates. METHODS: This study was part of an ongoing, multiclinic site, cervical cancer prevention trial in which patients completed a baseline survey, available in both Spanish and English, with the question: "Has a doctor or other healthcare provider ever asked you about domestic violence?" as well as other questions. RESULTS: Of 2591 women, 1017 (39%) chose to complete the survey in Spanish and 1574 (61%) in English. Within the entire group, 1137 (44%) reported having been asked about domestic violence. Among those completing the Spanish survey, this rate was 47% (lifetime assessment), and among English-language respondents, it was surprisingly lower at 42% (p=0.011). In multivariate analyses, however, this language effect was reduced to nonsignificance. Instead, age (particularly the 28-34-year quartile), having been pregnant, clinic site, and type of medical visit (postpartum) were positively associated with lifetime assessment. CONCLUSIONS: This study found a Spanish language preference is not a barrier to domestic violence assessment. PMID- 21631374 TI - Effect of chemical substances in removing methylene blue after photodynamic therapy in root canal treatment. AB - BACKGROUND AND OBJECTIVE: The disadvantage of photodynamic therapy (PDT) is in the photosensitizing agents that may stain the tooth structure. There is no register of PDT studies evaluating protocols to minimize that concern. The present study evaluated the efficiency of chemical adjuncts in methylene blue dye (MB) removal after PDT. MATERIALS AND METHODS: Forty single-rooted teeth, after root canal preparation, were filled with 0.01% MB for 5 min, and irradiated with diode laser 660 nm at 40 mW for 240 sec (total energy 9.6 J). The specimens were divided into four groups (n = 10), according to the chemical adjuncts used for dye removal: (a) 2.5% sodium hypochlorite (NaOCl); (b) 2.5% NaOCl + Endo-PTC cream; (c) 70% ethyl alcohol and (d) saline (control). The crowns were sectioned and fixed in a device. Photographs were taken before the PDT (T0), immediately after (T1) and upon dye removal (T2). The chromatic alterations were evaluated using Adobe Photoshop and K values were determined in four fixed points of each crown. RESULTS: K values (dental staining) increased in all groups when comparing T0 and T1. The effectiveness of the tested adjuncts was, in decreasing order: G1 (-3.11) > G2 (-2.97) > G3 (-1.28) > G4 (-1.19), not observing significant statistical differences (p < 0.05) between G1 and G2 and also between G3 and G4. CONCLUSIONS: Protocols to remove photosensitizing dyes should be applied after PDT in order to minimize dental stain. The protocols tested in this study by using 2.5% NaOCl, associated or not with Endo-PTC cream, were effective in avoiding tooth staining caused by MB during PDT. PMID- 21631375 TI - Influence of superpulsed laser therapy on healing processes following tooth extraction. AB - OBJECTIVE: This research studied the effects of laser therapy on healing processes following tooth extraction in healthy human subjects, evaluating some inflammation, osteogenesis, and clinical parameters. BACKGROUND DATA: Alveolar healing following tooth extraction is a complex repair process involving different types of tissues, including epithelium and bone. Therefore, it can be advantageous to use techniques able to influence the healing of all tissues. PATIENTS AND METHODS: Ten healthy human subjects with indications for bilateral tooth extraction entered the split-mouth study. The subject/patient becomes his/her own control, thereby eliminating all individual differences in response to laser treatment. This consisted of: 904-nm laser, 33 W peak power, 30 KHz, 200 ns, average power 200 mW, illuminated area 1 cm(2), 200 mW/cm(2), 15 min, 180 J, 180 J/cm(2). In each patient, one post-extraction site was treated with laser radiation, whereas the other was left untreated as a control. Soft-tissue specimens were removed from the extraction site before tooth extraction (T0) and 7 days after from extraction (T7); expression of inflammatory and osteogenesis parameters was evaluated on these specimens. The clinical parameter "pain" was evaluated for each subject. RESULTS: Superpulsed laser irradiation prevented the increase of interleukin (IL)-1beta, IL-6, IL-10, and cyclooxygenase-2 (COX-2), and induced an insignificant increase in collagen at 7 days after extraction, versus levels on day of extraction; no changes were found in the other parameters examined. Patients reported less pain at the site treated with superpulsed laser irradiation than at the control site. CONCLUSIONS: This study suggests that superpulsed laser irradiation may be a treatment of choice for patients scheduled for tooth extraction, as it provides clinical efficacy, is safe and well tolerated, and is able to prevent inflammation. PMID- 21631376 TI - Evaluation of the effects of polarized light (lambda400-200 nm) on the healing of third-degree burns in induced diabetic and nondiabetic rats. AB - OBJECTIVE: The study evaluated, by light microscopy, the repair process on third degree burns on diabetic and nondiabetic rats, illuminated or not, with a polarized light (PL) source. BACKGROUND DATA: Burns are severe injuries that result in the loss of fluid and destruction of tissue, infection, and shock that may result in death. Diabetes mellitus is a public health problem that, being uncontrolled, causes severe disturbance to the body metabolism, including on wound healing. PL sources have been shown to be effective in improving healing in many situations. MATERIALS AND METHODS: Ninety male Wistar rats were divided into two groups (n=45): nondiabetic and diabetic. In one of the groups, diabetes mellitus was induced by streptozotocin. A third-degree burn, measuring 1.5*1.5 cm(2), was created in the dorsum of each animal. Phototherapy (lambda400-2000 nm, 10.2 or 20.4 J/cm(2)) started immediately after burning and was repeated daily until animal death (7, 14, and 21 days). Specimens were taken, processed, and stained with H&E and Sirius red and immunomarked with cytokeratin (CK) AE1/AE3. Descriptive analysis was performed by light microscopy. RESULTS: Animals subjected to phototherapy showed an acceleration of the repair, the dose of 10.2 J/cm(2) being the one that caused best results, including higher deposition of collagen, quicker inflammatory reaction, and improved revascularization. CONCLUSIONS: Our results suggest that the use of PL (10.2 J/cm(2)) improves the healing of third-degree burns on both diabetic and nondiabetic animals. PMID- 21631377 TI - Leave your sidearms not your ethics at the door. PMID- 21631378 TI - The effect of increasing fluence on the treatment of actinic keratosis and photodamage by photodynamic therapy with 5-aminolevulinic acid and intense pulsed light. AB - BACKGROUND AND OBJECTIVE: Photodynamic therapy (PDT) with 5-aminolevulinic acid (ALA) and intense pulsed light (IPL) is a relatively new combination for the treatment of actinic keratosis (AK) and photodamage. The objective of this study was to determine the effect of increasing the fluence of IPL on the outcome of patients with these skin conditions. METHODS: Patients (N = 24) were randomly assigned to five treatment treatment groups: control (IPL alone) and ALA with 20, 25, 40, and 50 J/cm(2) fluence of IPL. Each patient received a single treatment. ALA was applied twice and allowed to incubate 2 h before IPL irradiation. Results were evaluated 5-7 days and 8 weeks after treatment. Clearance of AK lesions was evaluated by counting lesions before and after treatment, and improvement in photodamage was assessed by comparing pre- and post-treatment photographs. Statistical evaluation was based at nonparametric tests with a cut off level at p < 0.10 and a confidence interval at 95%. RESULTS: Responses to treatment were greatest in patients who received ALA and IPL fluences of 40 and 50 J/cm(2). Responses were "marked" in 19% of the patients receiving 50 J and "moderate" in 19% of the patients receiving 40 J. Compared to the mean pre treatment AK grades, the mean post-treatment grades were 56% lower in the 50 J treatment group, 32% lower in the 25 J group, 50% lower in the 40 J group, 20% lower in the 20 J group, and 7% lower in the control group. Erythema, edema, crusts and erosion, and pain did not cause any patient to discontinue the study. CONCLUSION: AK clearance, but not photorejuvenation, appears to improve with increasing fluence at the ALA PDT-IPL levels used in this study without serious adverse effects. PMID- 21631379 TI - Photomedicine and LLLT literature watch. PMID- 21631380 TI - An exploratory analysis of the effects of a weight loss plus exercise program on cellular quality control mechanisms in older overweight women. AB - Obese older adults are particularly susceptible to sarcopenia and have a higher prevalence of disability than their peers of normal weight. Interventions to improve body composition in late life are crucial to maintaining independence. The main mechanisms underlying sarcopenia have not been determined conclusively, but chronic inflammation, apoptosis, and impaired mitochondrial function are believed to play important roles. It has yet to be determined whether impaired cellular quality control mechanisms contribute to this process. The objective of this study was to assess the effects of a 6-month weight loss program combined with moderate-intensity exercise on the cellular quality control mechanisms autophagy and ubiquitin-proteasome, as well as on inflammation, apoptosis, and mitochondrial function, in the skeletal muscle of older obese women. The intervention resulted in significant weight loss (8.0 +/- 3.9 % vs. 0.4 +/- 3.1% of baseline weight, p = 0.002) and improvements in walking speed (reduced time to walk 400 meters, - 20.4 +/- 16% vs. - 2.5 +/- 12%, p = 0.03). In the intervention group, we observed a three-fold increase in messenger RNA (mRNA) levels of the autophagy regulators LC3B, Atg7, and lysosome-associated membrane protein-2 (LAMP 2) compared to controls. Changes in mRNA levels of FoxO3A and its targets MuRF1, MAFBx, and BNIP3 were on average seven-fold higher in the intervention group compared to controls, but these differences were not statistically significant. Tumor necrosis factor-alpha (TNF-alpha) mRNA levels were elevated after the intervention, but we did not detect significant changes in the downstream apoptosis markers caspase 8 and 3. Mitochondrial biogenesis markers (PGC1alpha and TFAm) were increased by the intervention, but this was not accompanied by significant changes in mitochondrial complex content and activity. In conclusion, although exploratory in nature, this study is among the first to report the stimulation of cellular quality control mechanisms elicited by a weight loss and exercise program in older obese women. PMID- 21631381 TI - Evaluation of unconstrained monitoring technology used in the smart bed for u health environment. AB - OBJECTIVE: Since ubiquitous technology is an emerging paradigm in healthcare, we wanted to evaluate its feasibility in the healthcare area. As a first step, we evaluated the feasibility of the smart bed. In this study, a series of experiments were conducted to evaluate the smart bed. MATERIALS AND METHODS: Smart bed is a continuous ballistocardiogram monitoring device developed by Seoul National University. Eleven healthy subjects participated in the study. Each subject slept in the smart bed for two nights. To measure meaningful sleep periods, noisy sections were removed from the original signal. The subject's heart activity was obtained by automatic peak detection, using the AF2 peak detection algorithm. In this study, the feasibility of the smart bed was evaluated in terms of the meaningful sleep periods and the accuracy of automatic peak detection. RESULTS: On an average, 92.8% of sleep duration was meaningful. The accuracy of peak detection algorithm was also evaluated. The algorithm proved to have an accuracy of 95%; thus, we observe that 95% of the peak detection results were correct in comparison with the manual results. CONCLUSIONS: Based on the experimental results, we conclude that the smart bed can be a useful device for long-term monitoring in ubiquitous healthcare environments. PMID- 21631382 TI - Service design attributes affecting diabetic patient preferences of telemedicine in South Korea. AB - OBJECTIVE: Attempts to introduce telemedicine in South Korea have failed mostly, leaving critical questions for service developers and providers about whether patients would be willing to pay for the service and how the service should be designed to encourage patient buy-in. In this study, we explore patients' valuations and preferences for each attribute of telemedicine service for diabetes management and evaluate patient willingness to pay for specific service attributes. MATERIALS AND METHODS: We conducted a conjoint survey to collect data on patients' stated preferences among telemedicine service alternatives. The alternatives for diabetes-related service differed in 10 attributes, including those related to price, type of service provider, and service scope. To estimate the relative importance of attributes, patients' willingness to pay for each attribute, and their probable choice of specific alternatives, we used a rank ordered logit model. A total of 118 respondents participated in the survey. RESULTS: All 10 attributes significantly affected patients' valuations and preferences, and demographic and disease characteristics, such as existence of complications and comorbidities, significantly affected patients' valuations of the attributes. Price was the most important attribute, followed by comprehensive scope of service, the availability of mobile phone-based delivery, and large general-hospital provided services. CONCLUSIONS: The study findings have significant implications for adoption policy and strategy of telemedicine in diabetes management care. Further, the methodology presented in this study can be used to draw knowledge needed to formulate effective policy for adoption of the necessary technology and for the design of services that attract potential beneficiaries. PMID- 21631383 TI - Economic evaluations of the effects of longevity on telemedicine and conventional healthcare provision. AB - OBJECTIVE: Providing healthcare for the elderly is one of the expectations of a telemedicine system to reduce the isolation of the aging and enhance the quality of long-term care for the elderly. Based on the experiences in Taiwan, this research aimed at investigating the influences of longevity on the demand for health services through telecommunications and conventional face-to-face methods and to note the relationship between telemedicine and conventional health services. MATERIALS AND METHODS: Multiple regression analysis was undertaken. The sample covers the period from 1995 to 2004. Our results indicate that a 1% increase in the proportion of elderly relative to the population significantly increases the quantity demanded of telehealth by 305.73 and the quantity demanded of face-to-face health services by 1.99 * 107. On the relationship between longevity and telemedicine, the society begins to request telehealth when the elderly proportion is >2.06%. With current 9.5% elderly proportion, there is a potential to provide 2,274.86 telemedicine health services that are currently provided at only 954.9 and underprovided by 1.38 times. On the relationship between longevity and conventional health services, when the elderly proportion is >0.56%, the needs for face-to-face health services increase in the elderly percentage. With 9.5% elderly proportion, the estimated volume of health services via conventional medical care system is 1.54 * 108. The current quantity of such services is 1.11*108 and underprovided by 38%. Estimation results reveal that online and conventional health services are complementary for enhancing health status in an aging society. PMID- 21631384 TI - Automatic identification of apnea through acoustic analysis for at-home screening. AB - OBJECTIVE: Although many studies have analyzed breathing sounds in the diagnosis of obstructive sleep apnea syndrome, the recording of snoring sounds at home is hampered by the various background noises of daily life. Recordings also frequently include talking during sleep, which may infringe the privacy of patients. MATERIALS AND METHODS: A recording system used a bone conduction microphone to record snoring sounds. This microphone reduced background noise. A simple system transmitted recorded breathing sound data for screening at a hospital as envelope data instead of complete sound recordings, thereby decreasing data volume and protecting privacy. RESULTS: In periods in which blood oxygen levels (SpO2) were drastically decreased, the probability of apnea as deduced from the envelope curve of breathing sounds was consistent with SpO2 values. CONCLUSIONS: This method provides a basis for telemonitoring of sleep apnea syndrome. PMID- 21631385 TI - Enhancing the quality of life and preserving independence for target needs populations through integration of assistive technology devices. AB - Telehealth Test Bed-Quality of Life Studies is a research study investigating, testing, evaluating, and demonstrating technologies that have the potential to improve the quality of life for target populations, such as warriors in transition, veterans, individuals with physical and mental disabilities, and adults age 65 and older, who may require assistive technology devices to aid in maintaining or improving their quality of life. Thousands of soldiers who fought in Operation Iraqi Freedom and Operation Enduring Freedom have been wounded in action or have sustained injuries from noncombat accidents. Many of these injuries affecting military populations, as well as the general public, have resulted in amputations, traumatic brain injuries, and other physical or mental impairments. Depending on the severity of the injury, assistive technologies may be temporarily needed, or as a long-term solution, to regain and maintain normal daily functions. Saint Francis University's Center of Excellence for Remote and Medically Under-Served Areas developed an evaluation matrix comparing assistive technologies to identify devices that will improve or maintain the quality of life for these target populations. The integration of telehealth and telerehabilitation applications into patients' daily lives was examined to help improve home rehabilitation via access to healthcare specialties in rural and medically underserved settings. Researchers identified and tested assistive technology devices to be included in a self-sufficient living environment. The continuation of this research involves recruiting individuals to test and evaluate the functions of these commercially available technologies and to complete data collection surveys and questionnaires. The results are useful in selecting devices that will enhance or extend the quality of life of the target populations. PMID- 21631386 TI - Improving older adults' experience with interactive voice response systems. AB - BACKGROUND: Interactive voice response (IVR) systems use computer-based voice recognition and software algorithms to conduct human/computer interactions. In recent years, there has been a proliferation of IVR applications in business and healthcare. The available evidence suggests that older people have negative attitudes towards IVR and experience significant difficulties using these systems. OBJECTIVE: The goal of this project was to identify areas of difficulties in IVR use by older people and propose strategies for improvement. MATERIALS AND METHODS: During two focus groups, we examined older people's perceptions of IVR systems and the most common difficulties experienced by seniors in interacting with these systems. We also recorded their suggestions for improvement of IVR. RESULTS: Frequency and chi square analyses were performed on the focus groups data. Some of the difficulties reported by participants in this study were congruent with previous findings, but we also uncovered some additional problems, such as frustration for not being able to reach an operator, being asked to wait too long on hold, being unable to recover from mistakes, and an absence of shortcuts in the systems. In addition, significant number of participants indicated that they prefer a system that adjusts to them automatically as opposed to a system that allows for adjustment. CONCLUSION: Generally, our findings suggest that the poor acceptability of IVR systems by older people could be improved by designing IVR algorithms that detect difficulties during an ongoing IVR exchange and direct people to different algorithms adapted for each person. PMID- 21631387 TI - In vitro and in vivo characterization of nonbiomedical- and biomedical-grade alginates for articular chondrocyte transplantation. AB - Alginate is a key hydrogel for cartilage tissue engineering. Here, we systematically evaluated four biomedical- and two nonbiomedical-grade alginates for their capacity to support the in vitro culture and in vivo transplantation of articular chondrocytes. Chondrocytes in all ultrapure alginates maintained high cell viability. Spheres composed of biomedical-grade, low-viscosity, high mannuronic acid content alginate showed the lowest decrease in size over time. Biomedical-grade, low-viscosity, high-guluronic acid content alginate allowed for optimal cell proliferation. Biomedical-grade, medium-viscosity, high-mannuronic acid content alginate promoted the highest production of proteoglycans. When transplanted into osteochondral defects in the knee joint of sheep in vivo, empty spheres were progressively surrounded by a granulation tissue. In marked contrast with these observations, all alginate spheres carrying allogeneic chondrocytes were gradually invaded by a granulation tissue containing multinucleated giant cells, lymphocytes, and fibroblasts, regardless whether they were based on biomedical- or nonbiomedical-grade alginates. After 21 days in vivo, transplanted chondrocytes were either viable or underwent necrosis, and apoptosis played a minor role in their early fate. The individual characteristics of these alginates may be valuable to tailor specific experimental and clinical strategies for cartilage tissue engineering. PMID- 21631388 TI - Efficient differentiation of human embryonic and induced pluripotent stem cells into functional astrocytes. AB - Human high-grade gliomas (hHGG) remain a therapeutic challenge in neuro-oncology despite current multimodality treatments. We recently demonstrated that murine embryonic stem cell (mESC)-derived astrocytes conditionally expressing proapoptotic genes can successfully be used to induce apoptosis and tumor shrinkage of hHGG tumor in vitro and in an in vivo mouse model. The first step in the translation of these results to the clinical settings, however, requires availability of human embryonic stem cells (hESC)- and/or induced pluripotent cell (hiPSC)-derived astrocytes engineered to express proapoptotic genes. The potential for directed differentiation of hESCs and hiPSCs to functional postmitotic astrocytes is not fully characterized. In this study, we show that once specified to neuro-epithelial lineage, hiPSC could be differentiated to astrocytes with a similar efficiency as hESC. However, our analyses of 2 hESC and 2 hiPSC cell lines showed some variability in differentiation potential into astrocytic lineages. Both the hESC- and hiPSC-derived astrocytes appeared to follow the functional properties of mESC-derived astrocytes, namely, migration and tropism for hHGG. This work provides evidence that hESC- and hiPSC-derived cells are able to generate functionally active astrocytes. These results demonstrate the feasibility of using iPSC-derived astrocytes, a new potential source for therapeutic use for brain tumors and other neurological diseases. PMID- 21631389 TI - Development and characterization of rifampicin loaded floating microspheres. AB - Gastroretentive floating microspheres have a potential for enhancing the bioavailability and controlled delivery of drugs. The present study involves development of rifampicin floating microspheres in order to increase the gastric retention time. The microspheres were prepared by solvent evaporation technique and characterized for particle size, shape, zeta-potential, entrapment, and release kinetics. The developed systems were almost spherical in shape. The entrapment efficiency was found to be 86.34%. The percentage buoyancy after 8 hours was found to be 61.06. The prepared microspheres exhibited prolonged drug release in gastric medium and hence could be utilized for sustained delivery of anti-tubercular drugs. PMID- 21631390 TI - Caffeic acid phenethyl ester (CAPE): correlation of structure and antioxidant properties. AB - Caffeic acid phenethyl ester (CAPE), a plant polyphenolic concentrated in honeybee propolis, has been found to be biologically active in a variety of pathways. The aim of this study was to determine the antioxidant activity of CAPE using different methods such as total antioxidant activity by the thiocyanate method, 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid radicals, 1,1 diphenyl-2-picryl-hydrazyl free radicals, N,N-dimethyl-p-phenylenediamine dihydrochloride radicals and superoxide anion radicals scavenging activities, reducing power and ferrous ions (Fe(2+)) chelating activities. CAPE showed 97.9% inhibition on lipid peroxidation of linoleic acid emulsion. On the other hand, butylated hydroxyanisole, butylated hydroxytoluene, alpha-tocopherol and trolox indicated an inhibition of 87.3, 97.6, 75.3 and 90.3% on peroxidation in the same system, respectively. PMID- 21631391 TI - Within-subject biological variation of glucose and HbA(1c) in healthy persons and in type 1 diabetes patients. AB - BACKGROUND: Several articles describing within-subject biological variation of fasting glucose and HbA(1c) in healthy populations have been published, but information about biological variation of glucose and HbA(1c) in patients with type 1 diabetes is scarce. It is reasonable to assume that type 1 diabetics differ from their healthy counterparts in this matter. The aim of our study was to estimate the biological variation of glucose and HbA(1c) in healthy subjects and in patients with type 1 diabetes. METHODS: Fifteen healthy individuals and 15 type 1 diabetes patients were included. Biological variations were calculated based on blood samples collected weekly for 10 consecutive weeks from the healthy and the eligible of the type 1 diabetes patients. RESULTS: The within-subject variations of glucose were approximately 5% in healthy individuals and 30% in diabetes patients, and for HbA(1c) they were 1.2% in healthy individuals and 1.7% in diabetes patients. CONCLUSIONS: In conclusion, we found a high within-subject biological variation of glucose in diabetes patients as expected compared to healthy individuals (30% vs. 5%). The short-term (2 months) within-subject biological variation of HbA(1c) did not differ significantly between well regulated type 1 diabetes patients and healthy individuals (1.7% vs. 1.2%). PMID- 21631392 TI - Prospective study of first stroke in relation to plasma homocysteine and MTHFR 677C>T and 1298A>C genotypes and haplotypes - evidence for an association with hemorrhagic stroke. AB - BACKGROUND: Abnormalities in homocysteine metabolism have been suggested as risk factors for stroke. The aim of this prospective study was to examine whether total plasma homocysteine concentration (tHcy) and its main genetic determinant, methylene tetrahydrofolate reductase (MTHFR) polymorphisms, were associated with first ischemic or hemorrhagic stroke. METHODS: This was a nested case-referent study of 321 ischemic and 60 hemorrhagic stroke cases, defined by WHO MONICA criteria and each matched with two event-free referents for sex, age, cohort, recruitment date and geographical area. All subjects were from the population based Northern Sweden Health and Disease Study cohorts. Odds ratios were determined by conditional logistic regression. RESULTS: The mean follow-up time was 4.2 years. Both tHcy and MTHFR were independent predictors of hemorrhagic stroke in multivariate models including body mass index, hypertension and, for MTHFR, tHcy [OR for the highest vs. lowest tHcy quartile 8.13 (95% CI 1.83-36.1), p(trend)=0.002; OR for MTHFR 677TT vs. 677CC genotype 3.62 (95% CI 0.77-17.0), p(trend)=0.040]. Haplotype analyses confirmed that the MTHFR 677T-1298A haplotype was positively associated with hemorrhagic stroke [OR 1.81 (95% CI 1.09-3.00), p=0.022], whereas the MTHFR 677C-1298C haplotype was not significantly related to either hemorrhagic or ischemic stroke. Neither tHcy nor the MTHFR polymorphisms were significant predictors of ischemic stroke. CONCLUSION: Both elevated plasma homocysteine levels and the MTHFR 677T allele are indicators of increased risk of hemorrhagic stroke in the northern Swedish population. PMID- 21631393 TI - Paradoxical action of growth factors: antiproliferative and proapoptotic signaling by HGF/c-MET. AB - Hepatocyte growth factor (HGF)/mesenchymal-epithelial transition factor (c-MET) signaling is usually associated with the promotion of cellular growth and often with progression of tumors. Nevertheless, under certain conditions HGF can also act as an antiproliferative and proapoptotic factor and can sensitize various cancer cells, treated with anticancer drugs, to apoptosis. Not only HGF but also its various truncated forms as well as intracellular fragments of its membrane receptor, c-MET, may act as antiproliferative and proapoptotic factors toward various cells. This review focuses on different mechanisms responsible for such paradoxical action of the known typical growth factor. It also points toward the possibilities of usage of this information in anticancer therapy. PMID- 21631394 TI - Preparation and characterization of bioadhesive systems containing propolis or sildenafil for dental pulp protection. AB - PURPOSE: Binary polymeric systems containing poloxamer 407 (P407) and Carbopol 934P (C934P) were designed to deliver propolis extract (PE) or sildenafil citrate for the endodontic treatment (pulp protection). METHODS: Gelation temperature, rheology (flow), bioadhesion, and in vitro drug release of formulations were determined. RESULTS: Formulations showed thermoresponsive behavior, existing as a liquid at room temperature and gel at 34-37 degrees C. In addition, they exhibited pseudoplastic flow and low degrees of thixotropy or rheopexy. The greatest bioadhesion was noted in the formulation containing 20% P407 (w/w) and 0.10% C934P (w/w). PE release from formulation containing 15% P407 (w/w) and 0.25% C934P (w/w) was controlled by the phenomenon of relaxation of polymer chains. Moreover, sildenafil release from formulation containing 20% P407 (w/w) and 0.10% C934P (w/w) was controlled by Fickian diffusion. CONCLUSION: The data obtained on these formulations indicate a potentially useful role in the endodontic treatment (pulp protection) and suggest they are worthy of clinical evaluation. PMID- 21631395 TI - Fructans from aged garlic extract produce a delayed immunoadjuvant response to ovalbumin antigen in BALB/c mice. AB - Garlic (Allium sativum) is known for its innumerable biological activities including immunomodulation. Aged garlic extract (AGE), an odorless garlic preparation, has been shown to have superior immunomodulatory properties over raw garlic extract. Although garlic is a very rich source of fructans (17%, fresh weight basis), AGE contains only 0.22% of raw garlic fructans. Aged garlic fructans (AGF) have recently been shown to possess immunomodulatory activities in vitro. Natural adjuvants capable of eliciting better immune response of a model antigen are important in developing newer vaccines. In the present study, the adjuvant activity of AGF has been investigated in BALB/c mice using ovalbumin (OVA, 30 ug) as an experimental antigen. The body weights of animals did not change significantly indicating that the administration of garlic fructans is well-tolerated. AGF produce a significant humoral (serum IgG) response to OVA in BALB/c mice administered mucosally by either intranasal or oral route--a delayed response appearing on 50th day at a dose of 30 ug AGF by intranasal route. However, the serum IgG response was seen earlier on 35th day at a dose of 100 ug AGF by oral route. Higher concentrations of AGF (>50 ug) were inhibitory for adjuvant activity by intranasal administration. These observations indicate that AGF display immunoadjuvant activity for a test antigen though the humoral immune response is delayed. PMID- 21631396 TI - Pseudoephedrine inhibits T-cell activation by targeting NF-kappaB, NFAT and AP-1 signaling pathways. AB - Pseudoephedrine (PSE) is a stereoisomer of ephedrine that is commonly used as a nasal decongestant in combination with other anti-inflammatory drugs for the symptomatic treatment of some common pathologies such as common cold. Herein, we describe for the first time the effects of PSE on T-cell activation events. We found that PSE inhibits interleukin-2 (IL-2) and tumor necrosis factor (TNF) alpha-gene transcription in stimulated Jurkat cells, a human T-cell leukemia cell line. To further characterize the inhibitory mechanisms of PSE at the transcriptional level, we examined the transcriptional activities of nuclear factor kappa B (NF-kappaB), nuclear factor of activated T cells (NFAT), and activator protein-1 (AP-1) transcription factors and found that PSE inhibited NF kappaB-dependent transcriptional activity without affecting either the phosphorylation, the degradation of the cytoplasmic NF-kappaB inhibitory protein, IkappaBalpha or the DNA-binding activity. However, phosphorylation of the p65/RelA subunit was clearly inhibited by PSE in stimulated cells. In addition, PSE inhibited the transcriptional activity of NFAT without interfering with the calcium-induced NFAT dephosphorylation event, which represents the major signaling pathway for its activation. NFAT cooperates with c-Jun, a compound of the AP-1 complex, to activate target genes, and we also found that PSE inhibited both JNK activation and AP-1 transcriptional activity. These findings provide new mechanistic insights into the potential immunomodulatory activities of PSE and highlight their potential in designing novel therapeutic strategies to manage inflammatory diseases. PMID- 21631398 TI - Abnormalities of the penis in utero--hypospadias on fetal MRI. AB - OBJECTIVE: To demonstrate the visualization of penile abnormalities on fetal magnetic resonance imaging (MRI). METHODS: This retrospective study included five fetuses (25+0 to 31+6 gestational weeks) with penile abnormalities, positively depicted on fetal MRI, using prenatal ultrasonography (US) as a standard of reference. On MRI, the penis, as well as the scrotum and testicles, were evaluated. All fetal organs were reviewed to define penile abnormalities as isolated or in association with other anomalies. Furthermore, US and MRI findings were compared. RESULTS: Posterior hypospadias were demonstrated in all five fetuses, and abnormal testicular descent in two. Associated anomalies were present in all five fetuses on MRI, including abdominal/urogenital pathologies in four; brain pathologies in three; and craniofacial, cardiac, musculoskeletal, pathologies each in one fetus, and intrauterine growth retardation in one fetus. Compared to US, additional MRI findings were shown in four of five fetuses. CONCLUSIONS: Our MRI results demonstrate the visualization of fetal penile abnormalities and associated pathologies, which may provide information for perinatal management. MRI may show additional findings compared to prenatal US in certain cases. PMID- 21631399 TI - Effect of geometric challenges on cell migration. AB - Cellular infiltration and colonization of three-dimensional (3D) porous scaffolds is influenced by many factors. One of the major factors is the internal architecture presented to the cells. In this work, we have developed and validated a microfluidic device that presents a multitude of geometric challenges to cells, mimicking the architectural aspects and characteristics of 3D porous scaffolds in a two-dimensional arrangement. This device has been utilized to investigate the influence of varying channel widths, degrees of channel tortuosity, the presence of contractions or expansions, and channel junctions on the migration of NIH 3T3 mouse fibroblasts and human bone marrow-derived mesenchymal stromal cell (hMSCs). These two cell types were observed to have vastly different migration characteristics; 3T3 fibroblasts migrate as a collective cell front, whereas hMSCs migrate as single cells. This resulted in 3T3 fibroblasts displaying significant differences in migration depending on the type of geometrical constraint, whereas hMSCs were only influenced by channel width when it approached that of the length scale of a single cell. The differences in migration characteristics were shown to be related to the expression of the intercellular junction protein N-cadherin. We observed that 3T3 fibroblasts express higher levels of N-cadherin than hMSCs and that N-cadherin inhibition modified the migration characteristics of the 3T3 fibroblasts, so that they were then similar to that of hMSCs. The results of this study both confirm the utility of the device and highlight that differences in migration characteristics of different cell types can be deterministic of how they may respond to geometric constraints within porous tissue engineering constructs. PMID- 21631400 TI - Therapeutic targets for the management of peripheral nerve injury-induced neuropathic pain. AB - Neuropathic pain is a debilitating form of treatment resistant chronic pain and responds poorly to the clinically available therapies. Studies from animal models of neuropathic pain have led to understanding of its pathobiology which includes complex interrelated pathways leading to peripheral and central neuronal sensitization. Advancements in the elucidation of neuropathic pain mechanisms have revealed a number of key targets that have been hypothesized to modulate clinical status. The present review discusses these therapeutic targets including noradrenaline and 5-HT reuptake inhibitors; sodium, calcium and potassium channels; inhibitory and excitatory neurotransmitters; neuropeptides including bradykinin, tachykinin, cholecystokinin, neuropeptide Y, vasoactive intestinal peptide, and CGRP; pro-inflammatory cytokines; MAP kinases; PPAR gamma; Na(+)/Ca(2+) exchanger; nitric oxide; purinergic receptors; neuronal nicotinic receptors; cation-dependent chloride transporters; oxidative stress; matrix metalloproteinase and plasminogen activators; growth factors; transient receptor potential (TRP) channels; endocannabinoids; histamine receptors; dopamine; sigma receptors, beta adrenergic receptors, endothelins, and D-amino acid oxidase. The exploitation of these targets may provide effective therapeutic agents for the management of peripheral nerve injury-induced neuropathic pain. PMID- 21631401 TI - Dendrimers as novel systems for delivery of neuropharmaceuticals to the brain. AB - Most of the newly developed drugs fails to achieve sufficient bioavailability in to brain due to low water solubility and low permeability. Drug delivery systems are one method for achieving entry of molecules to their desired site of action within the body. Dendrimers are the customizable nanopolymers with uniform and well-defined particle size and shape. Dendrimers are of eminent interest for biomedical applications because of their ability to cross cell membranes. This potential pharmaceutical delivery system crosses the blood brain barrier (BBB) and other important target points. The high level of control over the dendritic architecture (size, branching density, surface functionality) make dendrimers ideal carriers in the field of brain drug delivery of anticancer, antiinflammatory, and antimicrobial agents. Examples of dendrimers such as poly(amidoamine) (PAMAM), poly(propylene imine) (PPI) and polyether-copolyester (PEPE), Glyco, PEGylated, peptide and pH dendrimers are of outmost significance. These dendrimers carry the drug molecules by physical interactions (encapsulation) or through chemical bonding (prodrug approach), while pH sensitive dendrimers are able to deliver drug molecules by alteration of ionic exchange in the brain microenvironment at the tumor site. Techniques employing dendrimers could be especially useful for drugs targeting to Alzheimer's and Prion's diseases. The present review should be of value to scientists who wish to work on the dendrimers for the delivery of molecules into the brain by systemic dosing. PMID- 21631402 TI - A dual mechanism linking NGF/proNGF imbalance and early inflammation to Alzheimer's disease neurodegeneration in the AD11 anti-NGF mouse model. AB - The neurotrophin Nerve Growth Factor (NGF) is essential for the maintenance and differentiation of basal forebrain cholinergic neurons. Since basal forebrain cholinergic neurons represent one major neuronal population affected and progressively degenerating in Alzheimer's disease (AD), interest has grown for NGF as a potential therapeutic agent in neurodegenerative disorders linked to aging, particularly for AD. However, no evidence was available, to link, in a cause-effect manner, deficits in NGF signalling to the broader activation in the Alzheimer's cascade, besides cholinergic deficits. The phenotypic analysis of the AD11 anti-NGF transgenic mouse, obtained by the "neuroantibodies" phenotypic protein knock out strategy, allowed demonstrating a direct causal link between NGF deprivation and AD pathology. Since then, extensive mechanistic studies on the AD11 model provided a new twist to the concept that alterations in NGF transport and signalling play a crucial role in sporadic Alzheimer's neurodegeneration, leading to the hypothesis of "Neurotrophic imbalance" as an upstream driver for sporadic AD. The results obtained with the AD11 anti-NGF mice highlight the fact that the particular mode of NGF neutralization, with an NGF antibody expressed in the brain, selectively interfering with mature NGF versus unprocessed proNGF, plays a major role in the mechanism of neurodegeneration, and could lead to new insights into the mechanisms of human sporadic AD. Here, we will review (1) the renewed neurotrophic imbalance hypothesis for AD and (2) the mechanisms underlying the neurodegenerative phenotype of AD11 anti-NGF mice. PMID- 21631403 TI - The value of repetitive transcranial magnetic stimulation (rTMS) for the treatment of anxiety disorders: an integrative review. AB - Unlike for depression, only few studies are available today investigating the therapeutic effects of repetitive transcranial magnetic stimulation (rTMS) for anxiety disorders. This review aims to provide information on the current research approaches and main findings regarding the therapeutic use of rTMS in the context of various anxiety disorders. Although positive results have frequently been reported in both open and randomized controlled studies, our review of all identified studies indicates that at present no conclusive evidence of the efficacy of rTMS for the treatment for anxiety disorders is provided. Several treatment parameters have been used, making the interpretation of the results difficult. Moreover, sham-controlled research has often been unable to distinguish between response to rTMS and sham treatment. However, there is a limitation in the rTMS methods that likely impacts only the superficial cortical layers. It is not possible to directly stimulate more distant cortical areas, and also subcortical areas, relevant to the pathogenesis of anxiety disorders, though such effects in subcortical areas are thought to be indirect, via trans-synaptic connections. We thus recommend further studies to clearly determine the role of rTMS in the treatment of anxiety disorders. Key advances in combining TMS with neuroimaging technology may aid in such future developments. PMID- 21631404 TI - Neuroinflamm-aging and neurodegenerative diseases: an overview. AB - Neuroinflammation is considered a chronic activation of the immune response in the central nervous system (CNS) in response to different injuries. This brain immune activation results in various events: circulating immune cells infiltrate the CNS; resident cells are activated; and pro-inflammatory mediators produced and released induce neuroinflammatory brain disease. The effect of immune diffusible mediators on synaptic plasticity might result in CNS dysfunction during neuroinflammatory brain diseases. The CNS dysfunction may induce several human pathological conditions associated with both cognitive impairment and a variable degree of neuroinflammation. Furthermore, age has a powerful effect on enhanced susceptibility to neurodegenerative diseases and age-dependent enhanced neuroinflammatory processes may play an important role in toxin generation that causes death or dysfunction of neurons in neurodegenerative diseases This review will address current understanding of the relationship between ageing, neuroinflammation and neurodegenerative disease by focusing on the principal mechanisms by which the immune system influences the brain plastic phenomena. Also, the present review considers the principal human neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis and psychiatric disorders caused by aging and neuroinflammation. PMID- 21631405 TI - Neuronal nicotinic acetylcholine receptors: emerging therapeutic targets for alcoholism. Commentary. PMID- 21631406 TI - Pain--unmet need and emerging targets & therapies. PMID- 21631407 TI - Cannabinoid 1 (CB1) receptor--pharmacology, role in pain and recent developments in emerging CB1 agonists. AB - Cannabinoids are antinociceptive in animal models of acute pain, tissue injury and nerve injury induced nociception and act via their cognate receptors, cannabinoid receptor 1 and 2. This review examines the underlying biology of the endocannabinoids and behavioural, neurophysiological, neuroanatomical evidence supporting the notion of pain modulation by these ligands with a focus on the current evidence encompassing the pharmacological characterization of CB1 agonists in this therapy. Separating the psychotropic effects of CB1 agonists from their therapeutic benefits is the major challenge facing researchers in the field today and with the discovery of peripherally acting agonists there seems to be a ray of hope emerging for the diverse potential therapeutic applications of this class of ligands. PMID- 21631408 TI - Control of neuropathic pain by immune cells and opioids. AB - Neuropathic pain is a compilation of somatosensory, cognitive and emotional alterations developing following nerve injuries. Such pain often outlasts the initial cause and becomes a disease of its own that challenges its management. The actions of currently used anticonvulsants, antidepressants and opioids are hampered by serious central nervous system adverse effects, which preclude their sufficient dosing and long-term use. Conversely, selective activation of opioid receptors on peripheral sensory neurons has the advantage of pain relieve without central side effects. Considerable number of animal studies supports analgesic effects of exogenously applied opioids acting at peripheral opioid receptors in neuropathic conditions. In contrast to currently highlighted pain-promoting properties of neuroimmune interactions associated with neuropathy, recent findings suggest that opioid peptide-containing immune cells that accumulate at damaged nerves can also locally alleviate pain. Future aims include the exploration of opioid receptor signaling in injured nerves and of leukocytic opioid receptor function in pain modulation, development of approaches selectively delivering opioids and opioid-containing cells to injured tissues and investigation of interactions between exogenous and leukocyte-derived opioids. These efforts should lay a foundation for efficient and safe control of neuropathic pain. This article comprehensively analyzes the consequences of nerve injury on the expression of peripheral opioid receptors and peptides, and the impact of these changes on opioid analgesia, critically discussing positive and negative findings. Further focus is on a dual character of immune responses in the control of painful neuropathies. PMID- 21631409 TI - Targeting interleukin-1beta for pain. AB - Interleukin-1beta (IL-1beta) has been implicated in many inflammatory and autoimmune diseases. Its role in pain, however, is under-appreciated. This may in part be due to the challenges involved in approaching the target from a therapeutic stand point. The scope of this brief review is to understand the direct and indirect roles of IL-1beta in contributing to different pain states including inflammatory and neuropathic pain, and discuss approaches to block IL 1beta production or IL-1beta-receptor interactions. PMID- 21631411 TI - Preclinical assessment of drug combinations for the treatment of pain: isobolographic and dose-addition analysis of the opioidergic system. AB - Opioid analgesics are the most frequently prescribed medications for the treatment of moderate or severe pain; however, their use is constrained by unwanted side effects. One therapeutic approach used to improve the side effect profile of opioids is the administration of a second drug in an opioid-containing mixture. Preclinical studies designed to predict the therapeutic potential of novel opioid-containing drug combinations are currently underway, and must rely on quantitative methods to assess their interactive effects. In this manuscript, an overview of isobolographic analysis is presented along with recent advances in isobolographic theory pertaining to drugs that differ in efficacy and to the statistical analysis of dose-addition. Next, studies using these analyses to assess the interactive effects of opioids and novel adjunct drugs, including selective COX-2 inhibitors, alpha(2)-adrenergic receptor agonists, delta-opioids, glutamate receptor antagonists and cannabinoid receptor agonists, are reviewed. Finally, comments on the future assessment of drug combinations for the treatment of pain-related disorders are made. PMID- 21631410 TI - Fatty acid amide hydrolase inhibitors--progress and potential. AB - Fatty acid amide hydrolase (FAAH) is responsible for hydrolysis of endocannabinoid, anandamide (AEA), and N-acyl ethanolamines such as palmitoylethanolamine (PEA) and N-oleoylethanolamide (OEA). Genetic deletion or pharmacological inactivation of FAAH shows site-specific elevation of AEA that plays a role in the modulation of pain and other neurodegenerative disorders. The review elaborates recent progress and current status of diverse structural classes of reversible and irreversible FAAH inhibitors. The discussion also addresses ligand-enzyme active site interactions and mechanism of enzyme inactivation, emerging approaches to novel FAAH inhibitors, and ongoing efforts to address gaps in therapeutic utility of FAAH inhibitors. PMID- 21631412 TI - K-Screen: a free application for high throughput screening data analysis, visualization, and laboratory information management. AB - High throughput screening (HTS) has emerged as an important technique for allowing researchers to rapidly profile very large numbers of chemicals against drug targets. As recent and future advances make HTS cheaper to perform on even larger scales, the amount of data that has to be processed, analyzed, and searched will only grow larger in size and harder for researchers to manually sift through. It is therefore an unavoidable requirement that institutions utilizing HTS technology will need to begin looking for effective solutions in the maturing area of laboratory information management systems like many other types of labs have already done. K-Screen is one such solution. Our initial goal with K-Screen was to have an integrated application environment that supported data analysis, management, and presentation so we could efficiently perform client requested screens and searches as well as generate detailed reports on the results of those. Previously, we had attempted but failed to locate an existing software suite that sufficiently addressed all our requirements. K-Screen is a web accessible application that offers the ability to host a large chemical structure library, process and store single-dose (primary) and dose response (secondary) screening data, perform searches based on screening results, plate coordinates, and structure, substructure and structure similarity. It uses heat maps and histograms to visualize screen or plate level statistics. Interfaces to external searches against PubChem and ZINC databases are also provided. We feel that these features make K-Screen a practical and effective alternative to other commercial or academic HTS LIMS systems. PMID- 21631413 TI - Essential attributes identified in the design of a Laboratory Information Management System for a high throughput siRNA screening laboratory. AB - In recent years high throughput screening operations have become a critical application in functional and translational research. Although a seemingly unmanageable amount of data is generated by these high-throughput, large-scale techniques, through careful planning, an effective Laboratory Information Management System (LIMS) can be developed and implemented in order to streamline all phases of a workflow. Just as important as data mining and analysis procedures at the end of complex processes is the tracking of individual steps of applications that generate such data. Ultimately, the use of a customized LIMS will enable users to extract meaningful results from large datasets while trusting the robustness of their assays. To illustrate the design of a custom LIMS, this practical example is provided to highlight the important aspects of the design of a LIMS to effectively modulate all aspects of an siRNA screening service. This system incorporates inventory management, control of workflow, data handling and interaction with investigators, statisticians and administrators. All these modules are regulated in a synchronous manner within the LIMS. PMID- 21631414 TI - Tracking and controlling everything that affects quality is the key to a quality management system. AB - Every laboratory has a need to track and control the variables that drive the quality of the results. However, each laboratory is unique and what one organization deems to be a critical process to track and control will likely differ from other organizations. Furthermore, there is more than just the end product or result that needs to be tracked and controlled. All of the intermediate products and resources play a significant role in producing the final product and each of these needs to be included in the LIMS. At a high level, this article will present ideas and opinions on the following topics in relation to implementing a LIMS process tracking and control system in a laboratory: The difference between tracking and controlling processes; What to track and control in the lab; The "product" of the laboratory; Preventing mistakes in a laboratory; Comprehensive software platform options; The value of seeing a system as opposed to imagining it; The use of barcodes in the laboratory; and an assessment on using the Risk Based Approach in deciding what to include in the tracking system. PMID- 21631415 TI - Challenges in small screening laboratories: implementing an on-demand laboratory information management system. AB - The Miami Project to Cure Paralysis, part of the University of Miami Miller School of Medicine, includes a laboratory devoted to High Content Analysis (HCA) of neurons. The goal of the laboratory is to uncover signaling pathways, genes, compounds, or drugs that can be used to promote nerve growth. HCA permits the quantification of neuronal morphology, including the lengths and numbers of axons. HCA of various libraries on primary neurons requires a team-based approach, a variety of process steps and complex manipulations of cells and libraries to obtain meaningful results. HCA itself produces vast amounts of information including images, well-based data and cell-based phenotypic measures. Documenting and integrating the experimental workflows, library data and extensive experimental results is challenging. For academic laboratories generating large data sets from experiments involving thousands of perturbagens, a Laboratory Information Management System (LIMS) is the data tracking solution of choice. With both productivity and efficiency as driving rationales, the Miami Project has equipped its HCA laboratory with an On Demand or Software As A Service (SaaS) LIMS to ensure the quality of its experiments and workflows. The article discusses how the system was selected and integrated into the laboratory. The advantages of a SaaS based LIMS over a client-server based system are described. PMID- 21631416 TI - Master data management: getting your house in order. AB - The availability of high-throughput techniques combined with more exploratory and confirmatory studies in small-molecule science (e.g., probe- and drug-discovery) creates a significant need for structured approaches to data management. The probe- and drug-discovery scientific processes start and end with lower throughput experiments, connected often by high-throughput cheminformatics, screening, and small-molecule profiling experiments. A rigorous and disciplined approach to data management ensures that data can be used to ask complex questions of assay results, and allows many questions to be answered computationally, without the need for significant manual effort. A structured approach to recording scientific experimental design and observations involves using a consistently maintained set of 'master data' or 'metadata'. Master data include sets of tightly controlled terminology used to describe an experiment, including both materials and methods. Master data can be used at the level of an individual laboratory or with a scope as extensive as a whole community of scientists. Consistent use of master data increases experimental power by allowing data analysis to connect all parts of the discovery life cycle, across experiments performed by different researchers and from different laboratories, thus decreasing the opportunity cost for making novel connections between results. Despite the promise of this increased experimental power, challenges remain in implementation and consistent use of master data management (MDM) techniques in the laboratory. In this paper, we discuss how specific MDM techniques can enhance the quality and utility of scientific data at a project, laboratory, and institutional level. We present a model for storage and exploitation of master data, practical applications of these techniques in the research context of small-molecule science, and specific benefits of MDM to small molecule screening aimed at probe- and drug-discovery. PMID- 21631417 TI - Safety and efficacy of thioridazine as salvage therapy in Indian patients with XDR-TB. AB - New drugs are desperately needed to combat XDR-TB as effective treatment involves at least four drugs to which the patient is sensitive or has never received in the past. Most Indian patients have received almost all second line drugs and have amplified resistance to most of the available drugs. Thioridazine has proven anti tuberculous effects in vitro and in vivo mouse models and we used this drug as salvage therapy in 4 Indian patients with XDR (near total drug resistance) with advanced disease. We found this drug to be well tolerated, even in this malnourished and ill patient population. It also resulted in clinical improvement in 3 of the 4 patients. Larger studies are being planned with this drug being added on to standardized or individualized XDR-TB regimens at an earlier stage. Because thioridazine has been used successfully for therapy of XDR-TB when in combination with antibiotics to which the patients were nonresponsive, the suggestion has been made that Thioridazine is eligible for patent as "New Use". PMID- 21631418 TI - Novel pharmacological therapies for cardiac hypertrophy and heart failure. PMID- 21631419 TI - Nitric oxide and protection against cardiac ischemia. AB - Nitric oxide (NO) is produced in almost all tissues and it exerts a variety of biological actions under both physiological and pathological conditions. It is synthesized by three distinct enzymes: endothelial (eNOS), neuronal (nNOS) and inducible (iNOS) nitricoxide synthases. NO is a cardioprotective mediator in powerful cardioprotective processes such as pre- and post-conditioning ischemia;they operate largely in a NO-dependent manner. However, the activity of different NOSs isoforms as well as, the bioavailability of NO can be affected by a variety of disease conditions (in particular diabetes) and pathological situations associated with significantly elevated levels of the pro-inflammatory cytokine tumor necrosis factor-alpha (TNF-alpha). These adversely affect NO signaling, as well as the efficacy and safety of treatments with NO or NO containing agents. PMID- 21631420 TI - Neuregulin1 as novel therapy for heart failure. AB - Neuregulin1 proteins (NRG1s) are epidermal growth factor (EGF) family members which are ligands for the ErbB receptor tyrosine kinases (RTKs). A decade of research has revealed that the NRG1-ErbB signaling is essential for the cardiac development and pivotal for maintaining the physiological function of the adult heart. The first evidence regarding the protective effect of the ErbB2 signaling in the adult heart came from clinical trials in breast cancer patients using Trastuzumab, a monoclonal antibody that blocks the ErbB2 receptor. The incidence of the New York Heart Association (NYHA) class III/IV heart failure increased five-fold in patients treated concurrently with chemotherapy drug doxorubicin and Trastuzumab compared to those treated with doxorubicin alone. Subsequent studies further show that stimulation of the ErbB2 signaling by NRG1s improves cardiomyocyte survival, growth and proliferation, maintains cardiac myofibril structure, counterbalances excessive beta-adrenergic signaling and promotes angiogenesis in the heart. Injections of recombinant NRG1s improve cardiac function in animal models with myocardial infarction, doxorubicin, viral infection or pacing-induced heart failure. Recent clinical trials show that NRG1s are effective for improving the cardiac function in heart failure patients. These results suggest that NRG1s may become a new drug for the treatment of heart failure. NRG1s stimulate RTKs. This is different from Beta-blockers, ACE inhibitors (Angiotensin-Converting Enzyme) and Angiotensin II receptor blockers which inhibit the excessive activation of G-protein coupled receptors (GPCRs). A clear understanding of how NRG1-ErbB signaling regulates cardiac function is essential for successful use of NRG1s for heart failure. Here, we review the current knowledge of the NRG1-ErbB signaling in the heart and discuss the potential use of NRG1s as novel therapy for heart failure. PMID- 21631422 TI - Mechanisms and inhibitors of apoptosis in cardiovascular diseases. AB - Apoptosis or progress of programmed cell death is a tightly regulated process which plays an important role in various cardiovascular diseases particularly in myocardial infarction, reperfusion injury, and heart failure. Over the past two decades, investigations of several pathways have broadened our understanding of programmed cell death. Many anti-apoptotic interventions have targeted ischemia reperfusion, however only a limited number have been considered at the chronic stage of heart failure. Endogenous inhibitors, caspase inhibitors, PARP-1 inhibitors, as well as various other agents have been implicated as anti apoptotic interventions. This review summarizes the apoptotic pathways associated with heart failure, discusses the current anti-apoptotic interventions available and reviews the clinical implications. PMID- 21631421 TI - Phosphoinositide-3 kinase signaling in cardiac hypertrophy and heart failure. AB - Heart failure, a major symptom in the progression of cardiac hypertrophy, is a critical risk factor for cardiac death. A large body of research has investigated cardioprotective mechanisms that prevent or minimize hypertrophy, identifying a variety of specific peptide hormones, growth factors, and cytokines with cardioprotective properties. Recent investigation of the downstream effector pathways for these growth factors has identified molecules involved in the progression of cardiac hypertrophy and heart failure, including phosphoinositide 3-kinase (PI3K), Akt and mammalian target of rapamycin (mTOR). Using genetically modified transgenic or knockout mice and adenoviral targeting to manipulate expression or function in experimental models of heart failure, several investigators have demonstrated that the PI3K-Akt pathway regulates cardiomyocyte size, survival, angiogenesis, and inflammation in both physiological and pathological cardiac hypertrophy. In this review, we discuss the reciprocal regulation of PI3K, Akt and mTOR in cardiomyocytes and their association with cardiac disease. PMID- 21631423 TI - Vitamin D therapy in cardiac hypertrophy and heart failure. AB - Vitamin D3 is made in the skin, modified in the liver to form 25(OH)D, and then further hydroxylated in the kidney to form the active hormone, 1,25 dihydroxyvitamin D3 (calcitriol). Calcitriol binds to and activates the vitamin D receptor (VDR), a nuclear receptor, to regulate numerous downstream signaling pathways in different cells and tissues. Emerging evidence suggests that VDR plays an important role in modulating cardiovascular, immunological, metabolic and other functions. Data from preclinical, epidemiological and clinical studies have shown that deficiency in VDR activation is associated with an increased risk for cardiovascular disease (CVD). Results from interventional trials using either nutritional vitamin D or VDR agonists (VDRAs) support the idea that VDR activation is beneficial for improving the underlying factors of CVD such as hypertension, endothelial dysfunction, atherosclerosis, vascular calcification, cardiac hypertrophy and progressive renal dysfunction. Furthermore, a majority of chronic kidney disease (CKD) patients die of CVD and VDRA therapy is associated with a survival benefit in both pre-dialysis and dialysis CKD patients. Most of the studies measured serum 25(OH)D as an indication for vitamin D deficiency, which does not truly reflect the VDR activation status. Although VDR plays an important role in regulating cardiovascular function and VDRAs may be potentially useful for treating CVD, at present VDRAs are not indicated for the treatment of CVD. PMID- 21631424 TI - Application of nanomedicine in cardiovascular diseases and stroke. AB - Nanomedicine is using nanotechnology in the medical application, which could range from medical use of nanomaterials in drug delivery and bio-imaging to development of nano-scale devices and sensors for diagnosis and therapy. Nanomedicine could also include methods to evaluate nanomaterials solely to ensure safe use through careful monitoring for potential toxicity. In this review, we will outline some of the potential uses of nanotechnology in different fields of medicine with special emphasis on cardiovascular diseases and stroke, based on pathophysiologic basis. We will also review some of the known nanomaterials that are already being utilized in diagnosis and treatment, commonly the FDA approved nanomaterials and others that have demonstrated to be promising in clinical applications.Finally, we will discuss the potential limitations of using nanotechnology in medical applications. Since nanomedicine is now emerging and still in development, this review is not intended as a comprehensive or conclusive overview of nanomedicine. Instead, we hope to provide examples of what are available currently, and to demonstrate the enormous potentials of nanomedicine in order to meet the unresolved needs and new challenges of medicine. PMID- 21631425 TI - Long-term Medication Adherence in Patients Receiving Antiretroviral Drug Therapy. AB - OBJECTIVE: A long-term medication adherence project was designed and implemented in an urban HIV clinic to address antiretroviral medication adherence. DESIGN AND METHODS: We conducted a prospective study of patients on long-term antiretroviral therapy. Referred patients were on antiretroviral agents at least six months and had two consecutive detectable viral loads. A standarized form was utilized to assess medication adherence, including patient report, practitioner assessed barriers, and pharmacy refill history. Individualized interventions were developed to accommodate patient needs. RESULTS: Seventy-eight patients met inclusion criteria for a total of 81 cases per study protocol. The majority of cases had an identifiable cause related to missed and/or mistimed doses. Following adherence interventions, 51 of the 81 cases (63%) experienced a successful outcome. In addition, 16 of the 27 cases (59%) without an identifiable cause became undetectable following intervention. CONCLUSIONS: This novel approach demonstrates that a proactive method for addressing barriers to long term medication adherence yields improved patient understanding and preservation of treatment regimens. PMID- 21631426 TI - Vertebral Lesions from AIDS-Related Kaposi's Sarcoma. AB - BACKGROUND: Kaposi's sarcoma is commonly described in HIV/AIDS patients but usually manifests as overt skin lesions or visceral involvement. Bone involvement, particularly vertebral, is uncommon, especially when there is no adjacent cutaneous lesion but a small number of cases have been reported. Unlike many other diseases associated with HIV, Kaposi's sarcoma can occur despite a normal CD4 count. CASE PRESENTATION: A 44 year-old HIV positive Nigerian man presented with a 20 day history of severe, worsening lumbar back pain, nearly three years after an earlier diagnosis of a single cutaneous lesion consistent with Kaposi's sarcoma, for which he received chemo-radiotherapy. Despite varying previous compliance with his anti-retroviral therapy, he was thought to be taking his medications at time of presentation and his CD4 count was 408 cells/mm(3). No other organ involvement was found but a pathological fracture was seen on magnetic resonance imaging affecting L1 vertebra. A CT-guided needle aspiration biopsy was performed and a histological diagnosis subsequently confirmed Kaposi's sarcoma. The patient was treated with further courses of radiotherapy but had little clinical improvement. Indeed, a follow-up MRI four months later showed new involvement of a further four vertebrae, fortunately in the absence of progressive focal neurology. CONCLUSION: Vertebral Kaposi's sarcoma is a rare diagnosis but can be accurately diagnosed with CT or MRI imaging in conjunction with a histological diagnosis. An immunosuppressed patient presenting with bone pain should be thoroughly investigated for Kaposi's sarcoma as modern chemotherapeutic agents alongside anti-retroviral therapy may delay or prevent further devastating complications such as spinal cord compression. PMID- 21631427 TI - The burden of metabolic diseases amongst HIV positive patients on HAART attending The Johannesburg Hospital. AB - South Africa has the highest prevalence of human immunodeficiency virus (HIV) infection in the world. The improved life expectancy, due to the recent introduction of highly active antiretroviral therapy (HAART), may lead to an increased health burden related to metabolic disorders, resulting in an increased pressure on health-care services. The main objective of this study was to determine the prevalence of hypertension, diabetes, obesity and dyslipidemia in a sample of HAART-treated HIV infected patients, attending an HIV clinic in the Gauteng province. This was a cross-sectional study of 304 HIV positive patients enrolled between January 2009 and March 2009, including patients aged 18 to 45 years, on HAART for more than one year. Hypertension prevalence was 19.1% (95% confidence interval (CI) 14.7-23.5): 23.9% in men and 17.7% in women (P=0.10). Diabetes was diagnosed in 4 women. Hypercholesterolemia (total cholesterol >5.00 mmol/mL) was found in 32.2% (95% CI 27.0-37.5), low HDL cholesterol (<1.20 mmol/mL) in 45.7% (95% CI 40.1-51.3) and elevated LDL cholesterol (>4.10 mmol/mL) in 9.5% (95% CI 6.2-12.8); these prevalences were not different between sexes, whereas hypertriglyceridemia (>2.25 mmol(mL) (15.8%, 95% CI 11.7-19.9) was significantly more frequent in men (28.4% versus 12.2%, P=0.002). TC and LDL-C were positively correlated with CD4+ cell count (r=0.13, P=0.03 and r=0.12, P=0.03). In this sample, the traditional risk factors for cardiovascular disease had a high prevalence, despite the young age of our patients. Women seemed to be at higher risk than man, unlike other HIV populations where these comparisons were made (Uganda, Italy and Norway). Obesity and lipid abnormalities, highly prevalent in the general population, also appeared related to HIV-infection and CD4+ cell count, presumably as a consequence of ART exposure. Further studies are needed in order to survey a population where HIV infection is turning into a chronic disease, with its complications. PMID- 21631428 TI - HIV-1 reactivation induced by apicidin involves histone modification in latently infected cells. AB - The existence of stable, transcriptionally silent human immunodeficiency virus (HIV) in latently infected cells represents a major obstacle to acquired immune deficiency syndrome (AIDS) therapy. Histone deacetylase (HDAC) can inhibit histone acetylation, resulting in HIV-1 provirus transcription silence. Apicidin, a widely used antiparasitic drug, exhibits antiparasitic activity by inhibiting HDAC. Using the latently infected A10.6 cell line, we describe the dose- and time dependent manner in which Apicidin reverses HIV-1 latency. We found that Apicidin can synergize with trichostatin A (TSA) to activate HIV-1 gene expression. Chromatin immunoprecipitation (ChIP) assay further indicates that Apicidin induces HIV-1 reactivation by increasing the acetylation levels of H3 and H4 at nucleosome 1 in HIV-1 long terminal repeats (LTR). Our research reveals a potent activator for reactivating latent HIV-1 and shows promise for HIV-1 therapy. PMID- 21631429 TI - Response to HAART in treatment-naive HIV-infected patients with a prior diagnosis of tuberculosis or other opportunistic infections. AB - We aimed to evaluate immunological, virological and clinical response to HAART, as well as all-cause mortality, in treatment-naive patients with a diagnosis of tuberculosis (TB) in the prior 6 months, compared to subjects with another AIDS defining illness (ADI) or event-free individuals in an open, prospective and multicenter hospital-based cohort of HIV-infected naive adults (2004-2008). All cause mortality rates were calculated by Cox regression models. Among 4407 patients, 2400 (54.5%) started HAART: 110 (4.6%) had had previous TB and 414 (17.3%) another ADI. Median time from TB diagnosis to inititation of HAART was 53 days (IQR: 25.75-83.25), and for other ADI was 22 days (IQR: 8-42). Overall, 151 (6.3%) patients developed a new ADI during follow-up; 63% reached virological suppression and 69.4% had increases of >=50 CD4+/ul, at 6 months. No statistically significant differences were found according to a previous history of TB or another ADI. Overall, 85 subjects died in 4031 person-years of follow-up with a mortality rate of 2.1 (95%CI: 1.7-2.6). When compared to subjects who started HAART in the absence of a previous ADI (HR 1), a prior diagnosis of an ADI other than TB was significantly associated with an increased risk of death. (HR 1.6; 95%CI: 1.1-2.3), but not a diagnosis of TB (HR 1.15; 95%CI: 0.5-2.5). In conclusion, a previous diagnosis of TB or another ADI before HAART did not compromise short-term virological and immunological response to treatment. A prior diagnosis of an ADI different to TB significantly increased all cause mortality. PMID- 21631430 TI - The circularly permuted yellow fluorescent protein cpYFP that has been used as a superoxide probe is highly responsive to pH but not superoxide in mitochondria: implications for the existence of superoxide 'flashes'. AB - The properties of a cpYFP [circularly permuted YFP (yellow fluorescent protein)] reported to act as a superoxide sensor have been re-examined in Arabidopsis mitochondria. We have found that the probe has high pH sensitivity and that dynamics in the cpYFP signal disappeared when the matrix pH was clamped by nigericin. In contrast, genetic and pharmacological manipulation of matrix superoxide had no detectable effect on the cpYFP signal. These findings question the existence of superoxide flashes in mitochondria. PMID- 21631431 TI - Mutations in the transmembrane and juxtamembrane domains enhance IL27R transforming activity. AB - Cytokines and their receptors regulate haemopoiesis by controlling cellular growth, survival and differentiation. Thus it is not surprising that mutations of cytokine receptors contribute to the formation of haemopoietic disorders, including cancer. We recently identified transforming properties of IL27R, the ligand-binding component of the receptor for interleukin-27. Although wild-type IL27R exhibits transforming properties in haemopoietic cells, in the present study we set out to determine if the transforming activity of IL27R could be enhanced by mutation. We identified three mutations of IL27R that enhance its transforming activity. One of these mutations is a phenylalanine to cysteine mutation at residue 523 (F523C) in the transmembrane domain of the receptor. The two other mutations identified involve deletions of amino acids in the cytoplasmic juxtamembrane region of the receptor. Expression of each of these mutant IL27R proteins led to rapid cytokine-independent transformation in haemopoietic cells. Moreover, the rate of transformation induced by these mutants was significantly greater than that induced by wild-type IL27R. Expression of these IL27R mutants also induced enhanced activation of JAK (Janus kinase)/STAT (signal transducer and activator of transcription) signalling compared with wild type. An activating deletion mutation of IL27R enhanced homodimerization of the receptor by a mechanism that may involve disulfide bonding. These transforming IL27R mutants displayed equal or greater transforming activity than bona fide haemopoietic oncogenes such as BCR-ABL (breakpoint cluster region-Abelson murine leukaemia viral oncogene homologue) and JAK2-V617F. Since IL27R is expressed on haemopoietic stem cells, lymphoid cells and myeloid cells, including acute myeloid leukaemia blast cells, mutation of this receptor has the potential to contribute to a variety of haemopoietic neoplasms. PMID- 21631432 TI - The Arabidopsis phi class glutathione transferase AtGSTF2: binding and regulation by biologically active heterocyclic ligands. AB - The plant-specific phi class of glutathione transferases (GSTFs) are often highly stress-inducible and expressed in a tissue-specific manner, suggestive of them having important protective roles. To date, these functions remain largely unknown, although activities associated with the binding and transport of reactive metabolites have been proposed. Using a sensitive and selective binding screen, we have probed the Arabidopsis thaliana GSTFs for natural product ligands from bacteria and plants. Uniquely, when overexpressed in bacteria, family members GSTF2 and GSTF3 bound a series of heterocyclic compounds, including lumichrome, harmane, norharmane and indole-3-aldehyde. When screened against total metabolite extracts from A. thaliana, GSTF2 also selectively bound the indole-derived phytoalexin camalexin, as well as the flavonol quercetin-3-O rhamnoside. In each case, isothermal titration calorimetry revealed high-affinity binding (typically Kd<1 MUM), which was enhanced in the presence of glutathione and by the other heterocyclic ligands. With GSTF2, these secondary ligand associations resulted in an allosteric enhancement in glutathione-conjugating activity. Together with the known stress responsiveness of GSTF2 and its association with membrane vesicles, these results are suggestive of roles in regulating the binding and transport of defence-related compounds in planta. PMID- 21631433 TI - A critical review of the contribution of eye movement recordings to the neuropsychology of obsessive compulsive disorder. AB - OBJECTIVE: Dysfunctions of saccadic and/or smooth pursuit eye movements have been proposed as markers of obsessive compulsive disorder (OCD), but experimental results are inconsistent. The aim of this paper was to review the literature on eye movement dysfunctions in OCD to assess whether or not saccades or smooth pursuit may be used to diagnose and characterize OCD. METHOD: Literature was searched using PubMed, ISI Web of Knowledge, and PsycINFO databases for all studies reporting eye movements in adult patients suffering from OCD. RESULTS: Thirty-three articles were found. As expected, eye movements of the patients with OCD were mostly assessed with simple oculomotor paradigms involving saccadic and/or smooth pursuit control. In contrast to patients with schizophrenia, however, patients with OCD only displayed rather unspecific deficits, namely slight smooth pursuit impairments and longer response latencies on antisaccade tasks. There was no relationship between these deficits and the severity of patients' symptoms. Interestingly, eye movements of the patients with OCD were almost never recorded during more complex cognitive tasks. CONCLUSION: As in schizophrenia and autism, eye movement recordings during more complex tasks might help to better characterize the cognitive deficits associated with OCD. Such recordings may reveal specific OCD-related deficits that could be used as reliable diagnostic and/or classification tools. PMID- 21631434 TI - Fetomaternal alloimmune thrombocytopenia: increasing awareness. PMID- 21631435 TI - A review of pathophysiology and current treatment for neonatal alloimmune thrombocytopenia (NAIT) and introducing the Australian NAIT registry. AB - Fetomaternal or neonatal alloimmune thrombocytopenia (NAIT) is a rare but serious condition associated with significant fetal and neonatal morbidity and mortality. The most useful predictor of severe disease is a history of a sibling with an antenatal intracranial haemorrhage. However, NAIT can occur during the first pregnancy and may not be diagnosed until the neonatal period. Antenatal treatment options include maternal intravenous immunoglobulin (IVIG) and corticosteroid treatment, fetal blood sampling (FBS) and intrauterine platelet transfusion (IUT) and early delivery. FBS (with or without IUT) can be used to direct and monitor response to therapy, and to inform mode and timing of delivery. However, this procedure is associated with significant risks, including fetal death, and is generally now reserved for high-risk pregnancies. This review highlights the current understanding of the epidemiology and pathophysiology of NAIT and summarises current approaches to investigation and management. It also introduces the newly established Australian NAIT registry. Owing to the relative rarity of NAIT, accruing sufficient patient numbers for studies and clinical trials at an institutional level is difficult. This national registry will provide an opportunity to collect valuable information and inform future research on this condition. PMID- 21631436 TI - Uterine transplantation: one human case followed by a decade of experimental research in animal models. AB - Uterine transplantation (UTx) aims to treat unconditional uterine factor infertility by replacing a non-functioning or non-existing uterus. After one attempt of UTx in the human 10 years ago, intensive research has been performed. The results of these specific studies on surgical technique, ischaemia reperfusion injury, immunosuppression and fertility are discussed. PMID- 21631437 TI - Early adverse perinatal complications in preterm growth-restricted fetuses. AB - INTRODUCTION: Growth-restricted fetuses are at increased risk of adverse perinatal outcome when compared to their normally grown counterparts. The additional risks associated with growth restriction in preterm fetuses are not well quantified, and this meta-analysis serves to address this uncertainty. MATERIALS AND METHODS: This is a meta-analysis and meta-regression of all relevant studies published since 1997 investigating perinatal outcome in preterm growth-restricted fetuses. RESULTS: Growth-restricted fetuses across all gestational ages were found to be at significant risk of having low Apgar scores at 5 min, sepsis, intracranial haemorrhage, intrauterine and neonatal death, necrotising enterocolitis and respiratory complications. Although this risk reduced as gestation increased, it remained amplified in growth-restricted fetuses when compared to normally grown fetuses. CONCLUSION: This large meta analysis for the first time quantifies the additional perinatal risks associated with preterm fetal growth restriction and may help counsel parents about the complications these fetuses face following birth. PMID- 21631438 TI - Emergency peripartum hysterectomy: a 10-year review at the Royal Hospital for Women, Sydney. AB - BACKGROUND: There appears to be a rise in the rate of emergency peripartum hysterectomy (EPH) in the developed world. AIMS: To determine the incidence, indications, risk factors, complications and management of EPH in our tertiary level teaching hospital, the Royal Hospital for Women (RHW) in Sydney, over the last decade. METHODS: A retrospective analysis was conducted of all cases of EPH performed at the RHW between the years 1999-2008 inclusive. EPH was defined as one performed after 20 weeks gestation for uncontrollable uterine bleeding not responsive to conservative measures occurring at any time after delivery but within the first 6 weeks post-partum. Cases were ascertained via our hospital obstetric database. RESULTS: There were 33 EPH among 38,998 births, a rate of 0.85 per 1000 births. Indications for EPH were morbid adherence of the placenta (54.8%), placenta praevia (19.4%), uterine atony (12.9%) and uterine rupture or cervical laceration (9.7%). A significant association between previous caesarean section (CS) and abnormal placentation was confirmed (P=0.011), especially for morbid adherence of the placenta (P=0.004). There was one maternal death. Maternal morbidity was significant, with disseminated intravascular coagulation and urinary tract injury among the most common complications. All women required blood transfusions, and over a quarter were admitted to the intensive care unit. CONCLUSIONS: In our series, abnormal placentation causing severe haemorrhage was the commonest indication for EPH. Previous CS is a risk factor for abnormal placentation and particularly for morbid adherence of the placenta. The morbidity associated with EPH is considerable. PMID- 21631439 TI - Rate of postdates induction using first-trimester ultrasound to determine estimated due date: Wellington Regional Hospital audit. AB - AIM: The aim of this audit was to examine the effect of using first-trimester (<14 weeks) ultrasound scan to determine EDD (US EDD) on the rate of induction for postdates pregnancies at Wellington Regional Hospital. METHODS: Women with singleton live pregnancies who had postdates (>=41 weeks) induction at Wellington Hospital during January 2009 to November 2009 were identified using a computerised database [Perinatal Information Management System (PIMS)]. The first trimester ultrasound images and reports for these women were retrieved and reviewed by a specialist in obstetric ultrasound. Only ultrasound studies that had technically satisfactory images at <14 weeks were included. RESULTS: A total of 329 women with a singleton live pregnancy were induced for postdates during the study period. Of these women, 50 (15.2%) were not >=41 weeks on PIMS EDD and therefore on the best available evidence should not have been induced for being postdates. Of the remaining 279 women, 158 had first-trimester scans available for review. Forty-three of 158 (27%) were <41 weeks when US EDD was used. CONCLUSIONS: The rate of postdates inductions at Wellington NRH could be decreased by 38% if induction was limited to women over 41-week gestation and by using US EDD as opposed to last menstrual period EDD. The use of early gestational scans (<14 weeks) to estimate EDD lowers the rate of postdates induction. This is very similar to the observed findings in literature. PMID- 21631440 TI - Pregnancy outcomes in the current era of cystic fibrosis care: a 15-year experience. AB - BACKGROUND: With improvement in clinical care and longer survival of patients with cystic fibrosis (CF), pregnancy has become commonplace. However, the impact of pregnancy on maternal health and fetal outcomes requires ongoing review. METHODS: A retrospective study of 20 pregnancies from 18 women with CF during the period 1995-2009 was performed. Changes in lung function, body mass index (BMI) and development of gestational diabetes were recorded. Fetal outcomes and maternal survival were examined, and the influence of pre-pregnancy parameters on outcomes was evaluated. RESULTS: Mean maternal age at pregnancy was 29+/-5 years with a mean pre-pregnancy forced expiratory volume in 1 s (FEV1) of 65.6+/-20.8% predicted. Eleven of 20 pregnancies had a pre-pregnancy FEV1 <60% predicted. During pregnancy, FEV1% predicted fell by 4.8% (CI 1.6-7.9), but recovered to baseline within 6 months post-partum. Mothers gained a mean weight of 7.6+/-3.2 kg, and gestational diabetes developed in 43% of women. All women delivered live births apart from one therapeutic abortion. Five infants were preterm, and two mature infants had low birth weight. Three mothers either died or required lung transplantation after pregnancy (range 2.5-8.0 years). FEV1 <60% predicted and BMI <20 kg/m(2) were significant predictors of fetal complications. CONCLUSION: Most women tolerated pregnancy well without major complications despite many having at least moderate lung function impairment. Pre-pregnancy FEV1 and BMI were important predictors of outcomes. PMID- 21631441 TI - Obstetric profiles of foreign-born women in Western Australia using data linkage, 1998-2006. AB - BACKGROUND: Despite Western Australia (WA) having the highest proportion of overseas-born residents of any Australian state, no previous study has examined the general patterns of obstetric health of foreign-born women in WA. AIMS: To examine the obstetric profiles of foreign-born women in WA using routinely collected perinatal data. METHODS: The records of 59,245 confinements to foreign born women were compared with those of 149,737 Australian-born, non-Indigenous women in WA between 1998 and 2006 using chi-square tests and ANOVA procedures. RESULTS: Foreign-born women were generally older, more likely to be married and have partners in highly skilled occupations, and were less likely to have private insurance or be teenage mothers. They were more commonly grand multiparae and were more likely to give birth at age 35 or older. On average, foreign-born women experienced increased risk of gestational diabetes, pre-labour rupture of membranes, failure to progress, fetal distress, perineal laceration and post partum haemorrhage. They were less likely to have an induced labour and more likely to use fetal monitoring. Instrumental delivery, episiotomy and caesarean sections varied with maternal region of origin. CONCLUSIONS: Several important differences in the obstetric profiles of foreign-born women were found. These differences have useful implications for obstetric services in culturally and linguistically diverse populations. Collection of further variables would also benefit the future provision of equitable and culturally appropriate care to diverse immigrant groups. PMID- 21631442 TI - Obstetric admissions to an integrated general intensive care unit in a quaternary maternity facility. AB - BACKGROUND: Monash Medical Centre (MMC) is a university-affiliated tertiary referral hospital in Melbourne, Victoria, Australia. The hospital has a large obstetric service and is the only quarternary obstetric unit in Victoria. The intensive care unit (ICU) is a busy 21-bed general unit with a broad casemix. While there is no designated state service obstetric ICU in Victoria, MMC ICU has increasingly tried to accept all obstetric patients referred, from both MMC and externally. AIM: To provide a local perspective on obstetric intensive care in Australia. METHODS: A retrospective audit of obstetric ICU admissions over 2 years. RESULTS: Sixty women were admitted, of whom 46 were postpartum. Twenty nine women were transferred from external sites. Mean maternal age was 30.7 years, mean gestational age 34.5 weeks and mean Acute Physiology and Chronic Health Evaluation (APACHE) version IIIj score 33. Obstetric haemorrhage was the most common admission diagnosis, followed by hypertensive spectrum disorders. Three women were admitted for induction of labour. Median length of stay was 35 h. Twenty-seven women (45%) required mechanical ventilation. No woman died in the ICU, although one died in hospital post-ICU discharge. No data were collected on neonatal outcomes. CONCLUSIONS: Critically ill obstetric patients can be managed successfully in a general ICU with obstetric input. It may be sensible to cluster these patients into units that are best equipped to deal with them, especially in the ante- and peripartum period. PMID- 21631443 TI - A review of the technique and complications from 2,012 cases of laparoscopically assisted vaginal hysterectomy at a single institution. AB - AIMS: To present our experience of modified laparoscopically assisted vaginal hysterectomy (LAVH) and to evaluate the surgical outcomes and complications. METHODS: Women with benign gynaecologic tumours that underwent a modified LAVH at the Samsung Medical Centre were analysed retrospectively. The technique is primarily a vaginal hysterectomy with a minor component of the laparoscopic procedures (stage 2 laparoscopic hysterectomy (LH)) and had two modifications (vaginal anterior colpotomy and McCall culdoplasty) from the standard technique. RESULTS: A total of 2012 LAVH procedures were performed from January 2000 to May 2008. The mean duration of the operations and the uterine weight were 102+/-32 min and 305+/-168 g, respectively. In 196 (9.7%) cases, the uterine weight was more than 500 g. Conversion to laparotomy was needed in 97 cases. Major intraoperative complications occurred in 45 cases (2.2%): bladder injury, 26 (1.29%); bowel injury, nine (0.45%); haemorrhage of major vessels, nine (0.45%); and ureteral injury, one (0.05%). Major long-term complications occurred in three cases: one fistula and two trocar site herniations. CONCLUSIONS: Stage 2 LH combined with modified vaginal anterior colpotomy and modified McCall culdoplasty is safe and effective for benign gynaecologic tumours and the prevention of post LAVH vaginal prolapse. PMID- 21631444 TI - Clinical manifestations in women with isolated fallopian tubal torsion; a rare but important entity. AB - BACKGROUND: The diagnosis of adnexal torsion can be difficult to make, especially in isolated fallopian tube torsion (FTT). Only small series and several case reports on isolated FTT have been published in the literature. AIMS: To demonstrate symptom presentations, objective findings and surgical outcomes in women with isolated FTT over a 12-year period at a tertiary hospital. METHODS: Using the International Classification of Disease, Ninth Revision, and Clinical Modification, we reviewed the clinical records of women with isolated FTT during 1996 and 2008 at our institute. RESULTS: Seventeen women, age 13-50 (mean 32) with surgically proven isolated FTT, were included. Five women (29.4%) had tubal ligation history. None of these cases were diagnosed before operation. Pain characteristics were variable; the onset was sudden in 10 (58.8%) and 17 (100%) complained of lower abdominal pain, but only six (35.3%) had peritoneal signs. Other clinical manifestations were as follows: nausea or vomiting in seven women (41.2%), lower urinary tract symptoms in four (23.5%) and fever in three (17.6%). All the women had a cystic adnexal mass on ultrasound. An enlarged tubal mass (5 14 cm) was found in all the cases at surgery. Eleven women (64.7%) underwent laparoscopy, and six (35.3%) laparotomy for salpingectomy. CONCLUSIONS: The diagnosis of isolated FTT is often not made before surgical intervention because clinical features are non-specific. However, it needs to be considered in women with acute lower abdominal pain with a unilateral cystic adnexal mass. PMID- 21631445 TI - Can misoprostol reduce blood loss in laparoscopy-assisted vaginal hysterectomy? AB - BACKGROUND: Intraoperative blood loss is a concern in the surgical treatment of myomatous uteri. Misoprostol causes the myometrium and isolated uterine arteries to contract and has blood-saving effects in myomectomy. AIM: To assess the efficacy of rectal misoprostol in reducing haemorrhage during laparoscopy assisted vaginal hysterectomy (LAVH). METHODS: Retrospective case-control study. Women who had undergone LAVH for leiomyoma were enrolled (n=117). Forty-nine women who used 400 MUg of misoprostol rectally 1 h before LAVH were compared with 68 women who did not. The surgical outcomes were compared statistically with Mann Whitney rank sum test, chi(2) test, or Fisher's exact test. RESULTS: The demographic variables were similarly distributed in both groups. There were no significant differences in the estimated blood loss, reduction in haemoglobin, operation time, or uterine weight between the two groups (P>0.05). The rates of operative complications (4.1 vs 10.3% for the misoprostol and control groups, respectively, P=0.21) were not different. There was no febrile morbidity (>38 degrees C) within 24 h of the misoprostol insertion. CONCLUSION: These data do not support the use of rectal misoprostol to reduce blood loss during LAVH. The pharmacoclinical effects of misoprostol in the uterus should be clarified. PMID- 21631446 TI - Analysis of factors predicting success of metformin and clomiphene treatment for women with infertility owing to PCOS-related ovulation dysfunction in a randomised controlled trial. AB - BACKGROUND: Metformin has failed to gain wide acceptance as a first-line treatment option for women with anovulatory infertility related to polycystic ovary syndrome. This study aimed to ascertain factors that predict fertility success with treatment that included metformin compared to standard (non metformin) treatment. METHODS: Randomised trial data analysis by logistic regression of factors likely to have a differential influence on the likelihood of success of metformin versus non-metformin treatment amongst women with ovulation dysfunction related to polycystic ovary syndrome. RESULTS: metformin versus those receiving placebo and those with lower BMI who received metformin were more likely to become pregnant than their lower BMI counterparts who received placebo (P=0.039). The subpopulation of women with BMI<=32 kg/m(2) had no factors showing a significantly different impact on the chance of pregnancy for women treated with metformin versus those receiving clomiphene treatment or combination metformin/clomiphene treatment versus clomiphene treatment. There were no significantly different effects of free testosterone, fasting insulin, duration of infertility or ultrasound appearance of polycystic ovaries in any treatment groups. CONCLUSION: This study provides preliminary evidence that BMI may be an important prognostic factor in response to metformin for women with ovulation dysfunction related to polycystic ovary syndrome, suggesting that women with a lower BMI may respond better to metformin treatment versus placebo amongst women with BMI>32 kg/m(2) . Individual patient data meta-analysis of existing randomised trials would clarify this further and would assess whether other factors might predict better response to metformin versus standard treatments. PMID- 21631447 TI - Emergency department presentations for problems in early pregnancy. AB - BACKGROUND: Many women with problems in early pregnancy such as vaginal bleeding or abdominal pain present to the emergency department (ED). AIM: To describe demographic, service delivery and diagnostic characteristics of women who presented to the ED with a problem in early pregnancy. METHODS: Data were reviewed for all electronically available ED presentations in 2008 in NSW, Australia according to diagnostic codes related to problems in early pregnancy (N=12,061). Descriptive statistics were used to illustrate relevant characteristics, and adjusted odds ratios were used to highlight the predictors of key service delivery outcomes. RESULTS: Women who presented to the ED for a problem in early pregnancy accounted for 1.2% of all ED presentations for women. The average age of women who presented to the ED for a problem in early pregnancy was 29.3 years, with 25% aged 35 years or older. Over a fourth (28%) of women presented to the ED on a weekend and over a third (37%) presented after-hours. Most (70%) women were seen according to their triage category, and the median length of stay in the ED was just under 4 h. One-fourth of women were admitted to hospital, which was 3.8 times more likely among women with an ectopic pregnancy. CONCLUSIONS: The findings of this research may be useful for hospitals and clinicians to review and improve their current service delivery models for women who present to the ED with a problem in early pregnancy. PMID- 21631448 TI - A common sleep disorder in pregnancy: restless legs syndrome and its predictors. AB - Restless legs syndrome (RLS) is common in the third trimester of pregnancy. In this case-control study, 22.5% of 211 women had RLS. The RLS cases had a history of growing pains (GP) more frequently than controls (P=0.042). A family history of GP (P=0.025) and RLS (P=0.018) was more frequent among cases than controls. RLS in pregnancy is predicted by family history of RLS and GP, and by childhood history of GP. PMID- 21631449 TI - Rapid hypnosis as an anaesthesia adjunct for evacuation of postpartum vulval haematoma. AB - Hypnosis can be a useful therapeutic adjunct to pharmacological analgesia or anaesthesia in obstetrics. However, it is rarely considered a primary anaesthetic technique and is seldom employed in the acute surgical setting. Few obstetricians and anaesthetists currently utilise this technique in their clinical practice. We present a case report of a 34-year-old woman who successfully underwent evacuation of a large vulval haematoma using the simple hypnosis technique of 'believed-in imagination' as the principal anaesthetic technique with only minimal adjunctive pharmacological analgesia. PMID- 21631450 TI - The first Australian experience of heterotopic grafting of cryopreserved ovarian tissue: evidence of establishment of normal ovarian function. AB - Cryostorage of reproductive potential, in the form of ovarian cortex, for young women about to undergo cytotoxic therapies has been offered clinically for some time. However, the prospects of re-establishing reproductive function using this tissue remain unclear. We now report reproducible follicular development, oocyte retrieval and embryo development following heterotopic grafting of cryopreserved ovarian cortex which had been stored for over 10 years. PMID- 21631451 TI - Intra-operative cystoscopy in gynaecological surgery: a brief overview. AB - Recent studies using universal cystoscopy have demonstrated a higher rate of urinary tract injury at major gynaecological procedures than previously thought. These injuries are associated with significant medical and medico-legal implications. The sensitivity of visual inspection is low and cystoscopic screening with intravenous dye improves detection. We review the evidence for intra-operative cystoscopy in gynaecology, with emphasis on rates of urinary tract injury and the use of intravenous contrast dye. PMID- 21631452 TI - Assessment of first sexual intercourse in young women with a history of childhood sexual abuse. AB - Women reporting childhood sexual abuse (CSA) that involved actual or attempted penetration may not identify this as their first sexual intercourse. Data were drawn from a population-based, prospective cohort study spanning adolescence to adulthood. CSA prior to age 16 and age of first sexual intercourse with a male were assessed retrospectively. More than half of women reporting CSA in the form of actual or penetrative abuse reported an age of first sexual intercourse at or beyond 16 years. Direct questioning about CSA is needed to accurately ascertain sexual history. PMID- 21631453 TI - Adenomyosis is a potential cause of recurrent implantation failure during IVF treatment. AB - Four women, who previously had undergone multiple unsuccessful in vitro fertilisation (IVF) cycles because of failure of implantation of good quality embryos, were identified as having coexisting uterine adenomyosis. Endometrial biopsies showed that adenomyosis was associated with a prominent aggregation of macrophages within the superficial endometrial glands, potentially interfering with embryo implantation. The inactivation of adenomyosis by an ultra-long pituitary downregulation regime promptly resulted in successful pregnancy for all women in this case series. PMID- 21631454 TI - Primary clear cell carcinoma of a paratubal cyst: a case report with literature review. PMID- 21631455 TI - Making Bengali Brick Lane: claiming and contesting space in East London. AB - Based on a recent empirical project on 'the Bengal diaspora', the paper explores the construction and contestation of meanings around the iconic East London street, Brick Lane. Taking the 2006 protests around the film Brick Lane as its starting point, the paper draws on original interviews conducted in 2008 with a range of Bengali community representatives, to examine the narratives of space, community and belonging that emerge around the idea of Brick Lane as the 'cultural heartland' of the British Bangladeshi community. By exploring the representation, production and contestation of 'social space' through everyday practices, the paper engages with and contests the representation of minority ethnic 'communities' in the context of contemporary multicultural London and examines the process of 'claiming' and 'making' space in East London. In so doing, the paper contributes to a critical tradition that challenges essentialising and pathologizing accounts of ethnic communities and racialized spaces, or that places them outside of broader social and historical processes - redolent, for example, in contemporary discussions about 'parallel lives' or 'the clash of civilizations'. By contrast, this paper views social space as made through movement and narration, with a particular emphasis on the social agency of local Bengali inhabitants and the multiple meanings that emerge from within this 'imagined community'. However, rather than simply stressing the unfinished and processual nature of spatial meanings, the paper insists on the historical, embodied and affective dimensions of such meaning making, and a reckoning with the broader social and political landscape within which such meanings take shape. The focus on Brick Lane provides an empirically rich, geographically and historically located lens through which to explore the complex role of ethnicity as a marker of social space and of spatial practices of resistance and identity. By exploring Bengali Brick Lane through its narratives of past, present and future, these stories attest to the symbolic and emotional importance of such spaces, and to their complex imaginings. PMID- 21631456 TI - Paths to citizenship? Public views on the extension of rights to legal and second generation immigrants in Europe. AB - This study uses variations in the legal-institutional frameworks of citizenship to explore cross-nationally public views about granting equal rights to legal immigrants and citizenship status to second-generation immigrants in 20 European countries. We link the literatures on citizenship regimes and attitudes toward immigrants to construct a conceptual model that is tested using ISSP data from 2003 and a set of matching contextual measures. Results from hierarchical linear regression analyses indicate that (1) opposition to the extension of rights to legal immigrants is augmented by shorter periods of required residency for naturalization and (2) granting citizenship status to second-generation immigrants is not sensitive to whether a regime consents or not to citizenship by birth. Net of individual and contextual controls, the findings also show that resistance to the expansion of rights to legal immigrants is higher in countries consenting to dual citizenship. Furthermore, our analyses reveal that Eastern European respondents do not differ significantly from their Western counterparts with respect to extending rights to either category of immigrants. These results are discussed in reference to the diversity of citizenship regimes in Europe and in light of the existing debates on harmonizing immigration policies. PMID- 21631457 TI - Dissenting families and social movement abeyance: the transmission of neo-fascist frames in postwar Italy. AB - What explains the re-emergence of social movements after abeyance? Based on interviews with activists who belonged to the Italian neo-fascist movement of the late 1960s to early 1980s, this article documents the preservation of a neo fascist mobilization potential after 1945 through the parent-child transmission of frames. This process involved learning through talk, action and text. Both the nature of family frames and their congruence with movement frames depended on whether parents were right-wing or non-partisan. Research on abeyance should include the family among institutions that uphold continuity between waves of contention in pluralist regimes. PMID- 21631458 TI - Ageing in a spectacular 'youth culture': continuity, change and community amongst older goths. AB - This article explores the continuing involvement in youth music and style cultures of older participants through examination of the case study of the goth scene. It does so in the context of a widespread neglect, until recently, of what happens to participants of 'youth cultures' as they move beyond adolescence and also of a growing consensus about the broadening of youth itself as a life course period. Drawing on recent work on older participants in other music and style related groupings, the article uses original qualitative research to examine the developing lives and identities of goths as they become older. Rather than regarding continuing participation as a simple extension of youth, the focus is on the ways participation accompanied and was reconciled with material, domestic and physical elements of developing adult lives. Through reference to the case study, I emphasize the ways the experience of ageing for long-term music and style culture participants can constitute a collective experience. PMID- 21631459 TI - A tainted trade? Moral ambivalence and legitimation work in the private security industry. AB - The private security industry is often represented - and typically represents itself - as an expanding business, confident of its place in the world and sure of its ability to meet a rising demand for security. But closer inspection of the ways in which industry players talk about its past, present and future suggests that this self-promotion is accompanied by unease about the industry's condition and legitimacy. In this paper, we analyse the self-understandings of those who sell security - as revealed in interviews conducted with key industry players and in a range of trade materials - in order to highlight and dissect the constitutive elements of this ambivalence. This analysis begins by describing the reputational problems that are currently thought to beset the industry and the underlying fears about its status and worth that these difficulties disclose. We then examine how security players seek to legitimate the industry using various narratives of professionalization. Four such narratives are identified - regulation, education, association and borrowing - each of which seeks to justify private security and enhance the industry's social worth. What is striking about these legitimation claims is that they tend not to justify the selling of security in market terms. In conclusion we ask why this is the case and argue that market justifications are 'closed-off' by a moral ambivalence that attaches to an industry trading in products which cannot guarantee to deliver the condition that its consumers crave. PMID- 21631460 TI - Economic advice as a vocation: symbioses of scientific and political authority. AB - Academic economists perform an important function in advising politicians and state bureaucrats, lending them epistemological authority. This creates a challenge of institutional design and of professional vocation, of how these experts can combine their commitment to scientific analysis with their commitment towards their governmental patrons. This article examines the case of anti-trust economics, in which government economists are encouraged to remain as academically engaged as possible, so that their advice will be - or appear to be unpolluted by political or bureaucratic pressures. Yet this ideal is constantly compromised by the fact that the economists are nevertheless government employees, working beneath lawyers. Max Weber's concept of a 'vocation' is adopted to explore this tension, and his two lectures, 'Science as a Vocation' and 'Politics as a Vocation' are read side by side, to consider this core dilemma of academic policy advisors. PMID- 21631461 TI - Laughing it off? Humour, affect and emotion work in communities living with nuclear risk. AB - Over the past two decades, an increasing number of risk researchers have recognized that risks are not simply objective hazards but that the meanings of risk are discursively negotiated, dynamic and embedded within the wider social relations that constitute everyday life. A growing interest in the complexity and nuances of risk subjectivities has alerted sociocultural researchers not only to what is said in a risk situation, but also to how it is said and to what is unsaid and even, in a particular context, unsayable; to the intangible qualities of discourse that communicate additional meanings. Humour is both an intangible and marks such intangible meanings, yet it has largely been ignored and insufficiently theorized by risk researchers. In this paper, we draw upon insights from the humour literature - suspending the belief that humour is inherently good - to analyse and theorize humour as a way of examining the meanings and functions of risk. We show how humour can both mask and carefully reveal affectively charged states about living with nuclear risk. As such, it helps risk subjects to live with risk by suppressing vulnerabilities, enabling the negotiation of what constitutes a threat, and engendering a sense of empowerment. We conclude that humorous talk can be serious talk which can enrich our understandings of the lived experience of risk and of risk subjectivities. PMID- 21631462 TI - The cultural currency of a 'good' sense of humour: British comedy and new forms of distinction. AB - Traditionally considered lowbrow art par excellence, British comedy has grown steadily in legitimacy since the 'Alternative Comedy Movement' of the early 1980s. Yet while there might be evidence of a transformation in British comic production, there is little understanding of how this has been reflected in patterns of consumption. Indeed, there is a remarkable absence of studies probing comedy taste in British cultural sociology, most notably in Bennett et al's (2009) recent and otherwise exhaustive mapping of cultural taste and participation. This paper aims to plug this gap in the literature by examining contemporary comedy taste cultures in Britain. Drawing on a large-scale survey and in-depth interviews carried out at the Edinburgh Festival Fringe, it argues that comedy now represents an emerging field for the culturally privileged to activate their cultural capital resources. However, unlike previous studies on cultural capital and taste, this research finds that field-specific 'comic cultural capital' is mobilized less through taste for certain legitimate 'objects' of comedy and more through the expression of rarefied and somewhat 'disinterested'styles of comic appreciation. In short, it is 'embodied' rather than 'objectified' forms of cultural capital that largely distinguish the privileged in the field of comedy. PMID- 21631463 TI - De novo mutations in epilepsy. PMID- 21631464 TI - Motor skill deficits in children with partial hearing. AB - AIM: We examined the effect of partial hearing, including cochlear implantation, on the development of motor skills in children (aged 6-12y). METHOD: Three independent groups of children were selected: a partial hearing group (n=25 [14 males, 11 females]; mean age 8y 8mo, SD 1y 10mo), a nonverbal IQ-matched group (n=27 [15 males, 12 females]; mean age 9y, SD 1y 6mo), and an age-matched group (n=26 [8 males, 18 females]; mean age 8y 8mo, SD 1y 7mo) from three schools with special units for children with partial hearing. All children with partial hearing had a bilateral hearing loss >60 decibels. Motor and balance skills were assessed using the Movement Assessment Battery for Children (MABC) and two protocols from the NeuroCom Balance Master clinical procedures. RESULTS: The mean standardized total MABC score of the children with partial hearing (95% confidence interval [CI] 71.8-88.7) was significantly lower than both the age matched (95% CI 95.8-111.4; p<0.01) and the IQ-matched (95% CI 87.6-103.0; p=0.03) comparison groups. The children with partial hearing had particular difficulties with balance, most notably during tests of intersensory demand. However, subgroup analyses revealed that the effect of cochlear implantation was clearly dependent on the nature of the task. INTERPRETATION: Children with partial hearing are at high risk of clinical levels of motor deficit, with balance difficulties providing support for conventional vestibular deficit theory. However, the effect of cochlear implantation suggests that other sensory systems may be involved. A broader ecological perspective, which takes into account factors external to the child, may prove a useful framework for future research. PMID- 21631465 TI - Pain, pain genetics, and 'next-generation' pain genetics. PMID- 21631466 TI - Inflamed gut mucosa: downstream of interleukin-10. AB - BACKGROUND: Interleukin-10 is a pleiotropic cytokine, whose main function is limitation and ultimately termination of immune responses. This is especially true for environmental interfaces such as the gastrointestinal tract. IL-10 acts as a key mediator for maintaining gut homeostasis. IL-10 knockout mice are well established as a genetic model for inflammatory bowel disease (IBD), and sequence variants in the IL-10 locus contribute to ulcerative colitis (UC). DESIGN: This review covers the significance of IL-10 signalling in the intestinal immune response both in health and disease. It explains the biological role of IL-10, its deregulation in IBD and its contribution to intestinal inflammation via endoplasmic reticulum stress response. RESULTS: Many IBD susceptibility genes have been discovered in the past years, linking fundamental biological systems, like innate and adaptive immunity, stress responses, autophagy and mucosal barrier to the pathogenesis of Crohn's disease (CD) and UC. IL-10 has long been known for its substantial role in regulating gut immunity, but its contribution to IBD was somewhat elusive. A recent study identified mutations in either IL-10 receptor subunits that are associated with early-onset enterocolitis, a severe phenotype of IBD. Other than genetic variants of IL-10 receptors, IL-10 and STAT3 genes are also associated with IBD, emphasizing the involvement of the IL-10 signalling cascade in the pathogenesis of CD and UC. CONCLUSIONS: The discovery of inherited deregulations in the IL-10 signalling cascade is not only considered the missing link between IL-10 and intestinal homeostasis, but also demonstrates how findings made in animal models help explaining human disease. PMID- 21631467 TI - Small artery dilation and endothelial markers in cardiovascular risk patients. AB - BACKGROUND: The use of methods based on reactive hyperaemia of small distal arteries to assess endothelial function (EF) is increasing; however, the mechanisms regulating vascular function in large and small arteries are probably different. We studied the correlations between the hyperaemia reactivity of small peripheral arteries determined by peripheral artery tonometry (PAT) and the levels of serum biomarkers of EF, inflammation and oxidation in patients with cardiovascular (CV) risk factors. METHODS: Four hundred and seven patients with intermediate CV risk were recruited into a cross-sectional study to examine whether soluble endothelial, inflammatory and lipid oxidative biomarkers correlate with small artery reactive hyperaemia index (saRHI) values, which were measured by PAT. RESULTS: A significant correlation was found between saRHI values and the concentrations of soluble E-selectin (sE-selectin) and soluble vascular cell adhesion molecule 1 (sVCAM-1). These correlations were stronger when only non-metabolic syndrome patients (46%) were analysed (r = -0.310, P < 0.0001; r = -0.264, P < 0.0001, respectively). In this subgroup, the oxidised low density lipoprotein/LDL (oxLDL/LDL) was also correlated with saRHI (r = -0.193, P = 0.009). A stepwise regression study showed that sE-selectin was the only biomarker significantly correlated with saRHI values (P < 0.0001). In multivariate linear regression analysis, this relationship was still strong when the main confounding covariates were taken into consideration. CONCLUSIONS: Elevated levels of sE-selectin and, to a smaller degree, sVCAM-1 and oxLDL/LDL are associated with lower postischemic reactivity in the small distal arteries. sE-selectin is the main determinant biomarker of saRHI as assessed by regression analysis. The presence of multiple risk factors weakens this association. PMID- 21631471 TI - Sex matters: evaluating sex and gender in migraine and headache research. AB - Significant sex differences exist in migraine and other headache disorders. Several hypotheses have been proposed to explain these differences, including fluctuations in sex hormones and receptor binding, genetic factors, differences in exposure to environmental stressors, as well as differences in response to stress and pain perception; but how valid are some of these findings and can we improve the quality of research in this field? It is notable that the preponderance of animal pain studies use male subjects to study a predominantly female disorder. Furthermore, with respect to headache and migraine sex differences, limited data have been derived from animal models. Additionally, although sex differences (based on the categorization of male vs female) may be more routinely evaluated in clinical headache research than in the basic science research, greater attention to potential differences across the life cycle of women (ie, premenopausal vs postmenopausal differences) and menstrual cycle is warranted. In this manuscript we define the differences between "sex" and "gender" and highlight the importance of their application and use in headache research. The enhanced recognition and implementation of attention to sex differences throughout the hormonal and life-cycle phase in both human and animal research will only help to strengthen and further our understanding of migraine and may help guide the direction of future headache research. PMID- 21631472 TI - Sex-related differences in epidemiological and clinic-based headache studies. AB - This manuscript discusses sex-related differences in headache prevalence, the symptoms and natural history of migraine, associated disability, and co-morbid disorders. The role of sex hormones is discussed with reference to the effects of hormonal events across the reproductive years and the specific effects of the menstrual cycle on migraine. Differences between the sexes were identified across all parameters reviewed. Future research should ensure that data are analyzed separately for men and women to ensure that differences between the sexes are identified. PMID- 21631474 TI - Migraine genes and the relation to gender. AB - Migraine is an episodic brain disorder that is characterized by recurrent attacks of severe unilateral headache that are accompanied by various neurological symptoms. In addition, many patients have what is called an aura with visual and sensory disturbances. The majority of patients are female, suggesting that female hormones play an important role in the pathophysiology of the disorder. The molecular mechanisms, however, underlying this female preponderance are not well understood. It can be expected that the field of genetics that aims at identifying genetic factors that cause migraine by lowering the threshold for attacks will unravel some of these mechanisms. The 3 best known migraine genes encode ion transporters and were identified in families with familial hemiplegic migraine (FHM), a rare subtype of migraine with aura. FHM gene mutations cause alterations in mechanisms that control and modulate the neurotransmitter balance in the brain. Transgenic mice knock-in with human pathogenic mutations that were shown to exhibit some migraine-relevant features were very helpful in dissecting molecular mechanisms of migraine and pointed to a central role for cortical glutamate. In addition, transgenic mice that overexpress human RAMP1 exist and exhibit an increased sensitivity to calcitonin gene-related peptide. Findings from genetic and animal experiments on gender differences in migraine are discussed. Recently, a role for glutamate also came forward from a genome-wide association study in common migraine. By deciphering genetic and pathogenic migraine pathways, it can be expected that in the near future we will better understand mechanisms behind the female preponderance in migraine. PMID- 21631473 TI - Childhood abuse and migraine: epidemiology, sex differences, and potential mechanisms. AB - Migraine and maltreatment are both common conditions that are more prevalent in women. Epidemiological evidence supports an association between childhood abuse and headache, as well as pain in general, although some controversy exists based on methodological concerns of studying the influence of remote, traumatic, stigmatizing events in an often depressed population. There is a growing scientific body of knowledge regarding the neurobiological effects of abuse on brain function and structure that suggest a possible role of early life stress in the pathogenesis of migraine, and a differential impact based on sex. Advances in our understanding of the basic mechanisms by which an adverse environment interacts with and changes the genome, may suggest new treatment strategies. PMID- 21631475 TI - Sex-related differences in animal models of migraine headache. AB - Trigeminal nerve-mediated pain disorders such as migraine, temporomandibular joint disorder, and classical trigeminal neuralgia are more prevalent in women than in men. Female laboratory animals also show greater responses to various nociceptive stimuli than male animals. However, current knowledge of migraine pathogenesis is based primarily on experimental studies conducted in male animals and lack of migraine research with female animals limits clinical relevance. Migraine is triggered by any alteration in the intrinsic or extrinsic milieu and women at reproductive age are continuously prone to waxing and waning effects of female sex hormones. The experimental approach to this problem is complex because the rodent estrous cycle differs from the human cycle, and because exogenous hormone replacement in ovariectomized females has its limitations. The existence of multiple estrogen receptors in the trigeminal system also presents a challenge. Estrogens do not seem to directly affect calcitonin gene-related peptide or 5-HT(1D) receptors in the trigeminal system. Nonetheless, 2 estrogen receptors activate MAPK/ERK signaling pathway that mediates nociceptive processing in trigeminal nucleus caudalis. In addition, estrogen enhances susceptibility to cortical spreading depression, the neurobiological event underlying migraine aura, which may be independent of the estrous cycle. Further studies in female animals are required to clarify mechanisms underlying sex differences with respect to fluctuating sex hormones, cortical spreading depression, and excitability of the trigeminovascular system. PMID- 21631476 TI - Mechanisms of pain modulation by sex hormones in migraine. AB - A number of pain conditions, acute as well as chronic, are much more prevalent in women, such as temporomandibular disorder (TMD), irritable bowel syndrome, fibromyalgia, and migraine. The association of female sex steroids with these nociceptive conditions is well known, but the mechanisms of their effects on pain signaling are yet to be deciphered. We reviewed the mechanisms through which female sex steroids might influence the trigeminal nociceptive pathways with a focus on migraine. Sex steroid receptors are located in trigeminal circuits, providing the molecular substrate for direct effects. In addition to classical genomic effects, sex steroids exert rapid nongenomic actions to modulate nociceptive signaling. Although there are only a handful of studies that have directly addressed the effect of sex hormones in animal models of migraine, the putative mechanisms can be extrapolated from observations in animal models of other trigeminal pain disorders, like TMD. Sex hormones may regulate sensitization of trigeminal neurons by modulating expression of nociceptive mediator such as calcitonin gene-related peptide. Its expression is mostly positively regulated by estrogen, although a few studies also report an inverse relationship. Serotonin (5-Hydroxytryptamine [5-HT]) is a neurotransmitter implicated in migraine; its synthesis is enhanced in most parts of brain by estrogen, which increases expression of the rate-limiting enzyme tryptophan hydroxylase and decreases expression of the serotonin re-uptake transporter. Downstream signaling, including extracellular signal-regulated kinase activation, calcium-dependent mechanisms, and cAMP response element-binding activation, are thought to be the major signaling events affected by sex hormones. These findings need to be confirmed in migraine-specific animal models that may also provide clues to additional ion channels, neuropeptides, and intracellular signaling cascades that contribute to the increased prevalence of migraine in women. PMID- 21631477 TI - Psychosocial factors of relevance to sex and gender studies in headache. AB - Sex and gender differences in humans are being increasingly recognized not only in experimental pain paradigms but also clinically. Women experience various chronic pain conditions such as headache more than men and evidence differences in pain threshold and pain tolerance experimentally. In addition to biological underpinnings, psychosocial factors such as gender and social role expectations, coping strategies, and affective variables likely contribute to observed sex- and gender-related differences in headache. The present narrative reviews and summarizes extant literature pertaining to these psychosocial factors. Gender and social role expectations and coping styles differ between men and women who experience headache and pain, in turn affecting differences in responding to pain. Epidemiologic findings that women have higher rates of headache-related disability and psychiatric comorbidity have not been replicated regularly among treatment-seeking headache samples. Awareness of these differences may stimulate further research and enhance therapeutic opportunities for headache patients. PMID- 21631478 TI - Effect of telcagepant on spontaneous ischemia in cardiovascular patients in a randomized study. AB - OBJECTIVE: The primary purpose of the study was to explore the safety and tolerability of telcagepant in patients with stable coronary artery disease. BACKGROUND: Triptans are effective acute anti-migraine drugs whose vasoconstrictive effects limit their use in patients at risk for adverse cardiovascular events. Telcagepant, a calcitonin gene-related peptide receptor antagonist, is being developed for the acute treatment of migraine. Antagonism of calcitonin gene-related peptide, which does not appear to cause vasoconstriction, may allow for treatment of migraine in patients with coronary artery disease. METHODS: In this randomized, double-blind, placebo-controlled, crossover study, patients with documented stable coronary artery disease were assigned to 1 of 2 treatment sequences: telcagepant then placebo, or placebo then telcagepant. In each treatment period, patients received 2 doses of telcagepant 300-mg or placebo 2 hours apart. They remained in the research center for 24 hours after receiving the first dose of each period, during which time continuous 12-lead ambulatory electrocardiographic (Holter) monitoring was performed. RESULTS: Twenty-eight patients were enrolled; all patients completed the study and were included in all analyses. Telcagepant was generally well tolerated. No laboratory or serious adverse experiences were reported, and no patient discontinued due to an adverse experience. There were no consistent treatment-related changes in laboratory, vital signs or electrocardiogram safety parameters. Three patients (2 after receiving placebo and 1 after receiving telcagepant) experienced ST segment depression during the study; none of these patients reported chest pain. CONCLUSIONS: Two doses of 300-mg telcagepant, administered 2 hours apart, did not appear to exacerbate spontaneous ischemia and were generally well tolerated in a small cohort of patients with stable coronary artery disease. Results of this study support further evaluation of telcagepant in patients with stable coronary artery disease. PMID- 21631479 TI - The prevalence of migraine and probable migraine in a Brazilian favela: results of a community survey. AB - OBJECTIVES: The objectives of the present study were to estimate the 1-year prevalence of primary headaches and the role of select socio-demographic aspects in a representative sample of adults living in a Brazilian shanty town. BACKGROUND: Some socio-demographic factors, such as marital status, income, education, and job status have been described in studies with contentious results. Nevertheless, few studies have assessed the prevalence of headache and the role of socio-demographic aspects in very low-income communities. METHODS: A cross-sectional, population-based study was undertaken. Door-to-door interviews with 383 people were conducted. Individuals were aged greater than 18 years, randomly selected from the "Paraisopolis" shanty town in Sao Paulo, Brazil. The degree of the association was calculated through prevalence ratios and adjusted with backward logistic regression by gender, age, and some socio-demographic factors, including living conditions. RESULTS: The estimated 1-year prevalence of headache, migraine, chronic migraine, and tension-type headache were 47% (CI 95%: 39.5-52.6%), 20.4% (CI 95%: 16.6-24.9%), 8.4% (CI 95%: 6.1-12.0%), and 6.2% (CI 95%: 3.3-9.8%), respectively. Migraine was more prevalent in women and among employed people. No other relationship was found. The overall prevalence of migraine and chronic migraine in this very low-income community were high and migraine was associated with gender and job status. CONCLUSION: The overall prevalence of migraine and chronic migraine in this very low-income community were high and tension-type headache was low. A paradox was noted in the employment status and income association, one would expect higher levels of migraine in a low-income population, but higher numbers were found in those employed vs unemployed. These findings will need to be replicated in other population samples. PMID- 21631480 TI - Botulinum toxin for the treatment of chronic migraine: the placebo effect. AB - Botulinum toxin A used to treat headache evokes prominent placebo effects and it is likely that these effects are solely responsible for its apparent effectiveness. PMID- 21631482 TI - Spontaneous resolution of acute headache by migration of fungus ball. PMID- 21631481 TI - Direct costs of preventive headache treatments: comparison of behavioral and pharmacologic approaches. AB - OBJECTIVES: This study provides preliminary data and a framework to facilitate cost comparisons for pharmacologic vs behavioral approaches to headache prophylactic treatment. BACKGROUND: There are few empirical demonstrations of cumulative costs for pharmacologic and behavioral headache treatments, and there are no direct comparisons of short- and long-range (5-year) costs for pharmacologic vs behavioral headache treatments. METHODS: Two separate pilot surveys were distributed to a convenience sample of behavioral specialists and physicians identified from the membership of the American Headache Society. Costs of prototypical regimens for preventive pharmacologic treatment (PPT), clinic based behavioral treatment (CBBT), minimal contact behavioral treatment (MCBT), and group behavioral treatment were assessed. Each survey addressed total cost accumulated during treatment (ie, intake, professional fees) excluding costs of acute medications. The total costs of preventive headache therapy by type of treatment were then evaluated and compared over time. RESULTS: During the initial months of treatment, PPT with inexpensive mediations (<0.75 $/day) represents the least costly regimen and is comparable to MCBT in expense until 6 months. After 6 months, PPT is expected to become more costly, particularly when medication cost exceeds 0.75$ a day. When using an expensive medication (>3 $/day), preventive drug treatment becomes more expensive than CBBT after the first year. Long-term, and within year 1, MCBT was found to be the least costly approach to migraine prevention. CONCLUSIONS: Through year 1 of treatment, inexpensive prophylactic medications (such as generically available beta-blocker or tricyclic antidepressant medications) and behavioral interventions utilizing limited delivery formats (MCBT) are the least costly of the empirically validated interventions. This analysis suggests that, relative to pharmacologic options, limited format behavioral interventions are cost-competitive in the early phases of treatment and become more cost-efficient as the years of treatment accrue. PMID- 21631483 TI - The sexagenarian woman with new-onset cluster headaches. PMID- 21631484 TI - Migrainous complex hallucinations in a 17-year-old adolescent. AB - Olfactory hallucinations have been reported in association with numerous neurological and psychiatric disorders, in particular as a component of partial complex seizure and psychiatric disorders, but are rarely described in migraine disease. We report the case of an adolescent who reported complex hallucinations during a migraine attack. PMID- 21631485 TI - OnabotulinumtoxinA for treatment of chronic migraine. PMID- 21631487 TI - New daily persistent headaches: another traveling companion of the older headache doctor. PMID- 21631488 TI - A comment on brief sharp stabs of head pain and giant cell arteritis. PMID- 21631490 TI - Resident and fellow section. Teaching case: symptomatic SUNCT. PMID- 21631492 TI - Sex and headache. PMID- 21631494 TI - A double-blind placebo-controlled pilot study of sublingual feverfew and ginger (LipiGesicTM M) in the treatment of migraine. AB - BACKGROUND: Therapeutic needs of migraineurs vary considerably from patient to patient and even attack to attack. Some attacks require high-end therapy, while other attacks have treatment needs that are less immediate. While triptans are considered the "gold standard" of migraine therapy, they do have limitations and many patients are seeking other therapeutic alternatives. In 2005, an open-label study of feverfew/ginger suggested efficacy for attacks of migraine treated early during the mild headache phase of the attack. METHODS/MATERIALS: In this multi center pilot study, 60 patients treated 221 attacks of migraine with sublingual feverfew/ginger or placebo. All subjects met International Headache Society criteria for migraine with or without aura, experiencing 2-6 attacks of migraine per month within the previous 3 months. Subjects had <15 headache days per month and were not experiencing medication overuse headache. Inclusion required that subjects were able to identify a period of mild headache in at least 75% of attacks. Subjects were required to be able to distinguish migraine from non migraine headache. Subjects were randomized 3:1 to receive either sublingual feverfew/ginger or a matching placebo and were instructed but not required to treat with study medication at the earliest recognition of migraine. RESULTS: Sixty subjects treated 208 evaluable attacks of migraine over a 1-month period; 45 subjects treated 163 attacks with sublingual feverfew/ginger and 15 subjects treated 58 attacks with a sublingual placebo preparation. Evaluable diaries were completed for 151 attacks of migraine in the population using feverfew/ginger and 57 attacks for those attacks treated with placebo. At 2 hours, 32% of subjects receiving active medication and 16% of subjects receiving placebo were pain-free (P= .02). At 2 hours, 63% of subjects receiving feverfew/ginger found pain relief (pain-free or mild headache) vs 39% for placebo (P= .002). Pain level differences on a 4-point pain scale for those receiving feverfew/ginger vs placebo were -0.24 vs -0.04 respectively (P= .006). Feverfew/ginger was generally well tolerated with oral numbness and nausea being the most frequently occurring adverse event. CONCLUSION: Sublingual feverfew/ginger appears safe and effective as a first-line abortive treatment for a population of migraineurs who frequently experience mild headache prior to the onset of moderate to severe headache. PMID- 21631491 TI - Chemical mediators of migraine: preclinical and clinical observations. AB - Migraine is a neurovascular disorder, and although the pathophysiology of migraine has not been fully delineated, much has been learned in the past 50 years. This knowledge has been accompanied by significant advancements in the way migraine is viewed as a disease process and in the development therapeutic options. In this review, we will focus on 4 mediators (nitric oxide, histamine, serotonin, and calcitonin gene-related peptide) which have significantly advanced our understanding of migraine as a disease entity. For each mediator we begin by reviewing the preclinical data linking it to migraine pathophysiology, first focusing on the vascular mechanisms, then the neuronal mechanisms. The preclinical data are then followed by a review of the clinical data which support each mediator's role in migraine and highlights the pharmacological agents which target these mediators for migraine therapy. PMID- 21631496 TI - The immunological synapse: a cause or consequence of T-cell receptor triggering? AB - The immunological synapse forms as a result of the tight apposition of a T cell with an antigen-presenting cell (APC) and it is the site where the T-cell receptor (TCR) is triggered by its antigen ligand, the peptide-MHC complex present in the APC membrane. The immunological synapse was initially characterized in the T-cell membrane as three concentric rings of membrane receptors and their underlying cytoskeletal and signalling proteins. The inner circle, or central supramolecular activation cluster (cSMAC), concentrates most of the TCR and CD28, and it is surrounded by the peripheral SMAC that is formed by integrins. Finally, the most external ring or distal SMAC (dSMAC) is where proteins with large ectodomains are located, such as CD43 and CD45, far from the cSMAC. This arrangement was initially thought to be responsible for maintaining sustained TCR signalling, however, this typical concentric bull's-eye pattern is not found in the immunological synapses formed with the APCs of dendritic cells. Interestingly, TCR signalling has been detected in microclusters formed in the dSMAC area and it extinguishes as the TCRs reach the cSMAC. Hence, it appears that TCR signalling and full T-cell activation do not require the formation of the cSMAC and that this structure may rather play a role in TCR down-regulation, as well as participating in the polarized secretion of lytic granules. Here, we shall review the historical evolution of the role of the cSMAC in T-cell activation, finally discussing our most recent data indicating that the cSMAC serves to internalize exhausted TCRs by phagocytosis. PMID- 21631497 TI - Subversion of innate and adaptive immune activation induced by structurally modified lipopolysaccharide from Salmonella typhimurium. AB - Salmonella are successful pathogens that infect millions of people every year. During infection, Salmonella typhimurium changes the structure of its lipopolysaccharide (LPS) in response to the host environment, rendering bacteria resistant to cationic peptide lysis in vitro. However, the role of these structural changes in LPS as in vivo virulence factors and their effects on immune responses and the generation of immunity are largely unknown. We report that modified LPS are less efficient than wild-type LPS at inducing pro inflammatory responses. The impact of this LPS-mediated subversion of innate immune responses was demonstrated by increased mortality in mice infected with a non-lethal dose of an attenuated S. typhimurium strain mixed with the modified LPS moieties. Up-regulation of co-stimulatory molecules on antigen-presenting cells and CD4(+) T-cell activation were affected by these modified LPS. Strains of S. typhimurium carrying structurally modified LPS are markedly less efficient at inducing specific antibody responses. Immunization with modified LPS moiety preparations combined with experimental antigens, induced an impaired Toll-like receptor 4-mediated adjuvant effect. Strains of S. typhimurium carrying structurally modified LPS are markedly less efficient at inducing immunity against challenge with virulent S. typhimurium. Hence, changes in S. typhimurium LPS structure impact not only on innate immune responses but also on both humoral and cellular adaptive immune responses. PMID- 21631495 TI - Interleukin-23 and T helper 17-type responses in intestinal inflammation: from cytokines to T-cell plasticity. AB - Interleukin-23 (IL-23) plays an essential role in driving intestinal pathology in experimental models of both T-cell-dependent and innate colitis. Furthermore, genome-wide association studies have identified several single-nucleotide polymorphisms in the IL-23 receptor (IL-23R) gene that are associated with either susceptibility or resistance to inflammatory bowel disease in humans. Although initially found to support the expansion and maintenance of CD4(+) T helper 17 (Th17) cells, IL-23 is now recognized as having multiple effects on the immune response, including restraining Foxp3(+) regulatory T-cell activity and inducing the expression of Th17-type cytokines from non-T-cell sources. Here we focus on Th17 cells and their associated cytokines IL-17A, IL-17F, IL-21 and IL-22. We review studies performed in mouse models of colitis where these effector cytokines have been shown to have either a pathogenic or a tissue-protective function. We also discuss the heterogeneity found within the Th17 population and the phenomenon of plasticity of Th17 cells, in particular the ability of these lymphocytes to extinguish IL-17 expression and turn on interferon-gamma production to become Th1-like 'ex-Th17' cells. Interleukin-23 has been identified as a key driver in this process, and this may be an additional mechanism by which IL-23 promotes pathology in the intestinal tract. These 'ex-Th17' cells may contribute to disease pathogenesis through their secretion of pro-inflammatory mediators. PMID- 21631499 TI - The impact of a pathogenic bacterium on a social carnivore population. AB - 1. The long-term ecological impact of pathogens on group-living, large mammal populations is largely unknown. We evaluated the impact of a pathogenic bacterium, Streptococcus equi ruminatorum, and other key ecological factors on the dynamics of the spotted hyena Crocuta crocuta population in the Ngorongoro Crater, Tanzania. 2. We compared key demographic parameters during two years when external signs of bacterial infection were prevalent ('outbreak') and periods of five years before and after the outbreak when such signs were absent or rare. We also tested for density dependence and calculated the basic reproductive rate R(0) of the bacterium. 3. During the five pre-outbreak years, the mean annual hyena mortality rate was 0.088, and annual population growth was relatively high (13.6%). During the outbreak, mortality increased by 78% to a rate of 0.156, resulting in an annual population decline of 4.3%. After the outbreak, population size increased moderately (5.1%) during the first three post-outbreak years before resuming a growth similar to pre-outbreak levels (13.9%). We found no evidence that these demographic changes were driven by density dependence or other ecological factors. 4. Most hyenas showed signs of infection when prey abundance in their territory was low. During the outbreak, mortality increased among adult males and yearlings, but not among adult females - the socially dominant group members. These results suggest that infection and mortality were modulated by factors linked to low social status and poor nutrition. During the outbreak, we estimated R(0) for the bacterium to be 2.7, indicating relatively fast transmission. 5. Our results suggest that the short-term 'top-down' impact of S. equi ruminatorum during the outbreak was driven by 'bottom-up' effects on nutritionally disadvantaged age-sex classes, whereas the longer-term post outbreak reduction in population growth was caused by poor survival of juveniles during the outbreak and subsequent poor recruitment of breeding females. These results suggest synergistic effects of 'bottom-up' and 'top-down' processes on host population dynamics. PMID- 21631498 TI - The phenotypic and functional consequences of tumour necrosis factor receptor type 2 expression on CD4(+) FoxP3(+) regulatory T cells. AB - Cytokine receptors expressed by CD4(+) FoxP3(+) regulatory T cells (Treg cells) not only serve as a phenotypic marker for the identification of this important population of immunosuppressive cells, they also promote the function of Treg cells. CD25, the alpha-chain of interleukin-2 receptor, is a prototype of such a receptor, which enables Treg cells to be activated by interleukin-2. We and others have found that tumour necrosis factor receptor type 2 (TNFR2) is another important cytokine receptor preferentially expressed by Treg cells with important phenotypic and functional roles. TNFR2 is preferentially expressed by highly functional human and mouse Treg cells, and mediates the activating effect of TNF on Treg cells. We review here the studies of the regulation of expression of TNFR2 on functional Treg cells as well as on CD4(+) FoxP3(-) effector T cells (Teff cells). We document the critical role of this receptor in the activation, proliferative expansion and survival of Treg cells. The contribution of TNFR2 expression on Treg and Teff cells to the beneficial and detrimental effects of anti-TNF treatment in autoimmune disorders will also be discussed. PMID- 21631500 TI - Simulation of transmission electron microscope images of biological specimens. AB - We present a new approach to simulate electron cryo-microscope images of biological specimens. The framework for simulation consists of two parts; the first is a phantom generator that generates a model of a specimen suitable for simulation, the second is a transmission electron microscope simulator. The phantom generator calculates the scattering potential of an atomic structure in aqueous buffer and allows the user to define the distribution of molecules in the simulated image. The simulator includes a well defined electron-specimen interaction model based on the scalar Schrodinger equation, the contrast transfer function for optics, and a noise model that includes shot noise as well as detector noise including detector blurring. To enable optimal performance, the simulation framework also includes a calibration protocol for setting simulation parameters. To test the accuracy of the new framework for simulation, we compare simulated images to experimental images recorded of the Tobacco Mosaic Virus (TMV) in vitreous ice. The simulated and experimental images show good agreement with respect to contrast variations depending on dose and defocus. Furthermore, random fluctuations present in experimental and simulated images exhibit similar statistical properties. The simulator has been designed to provide a platform for development of new instrumentation and image processing procedures in single particle electron microscopy, two-dimensional crystallography and electron tomography with well documented protocols and an open source code into which new improvements and extensions are easily incorporated. PMID- 21631501 TI - Geometrical characterization of interconnected phase networks in three dimensions. AB - In electrochemical devices such as fuel cells or batteries the microstructure is a determining factor for the performance of the device. To be able to optimize the microstructure it is important to be able to quantitatively measure key structural parameters, such that systematic studies can be made. We present several general methods for quantitative characterization of network structures without prior assumptions of shape or application. The characterization is performed by extracting distributions of values rather than single value descriptions, thus allowing more detailed comparisons between samples to be made. The methods characterize tortuosity, path diameters, the novel dead ends property and a particle shape independent alternative to a particle size distribution. The parameters are calculated by the computation of arrival time maps by the fast marching method. The methods are applied to the analysis of each of the three phases in a solid oxide fuel cell sample. PMID- 21631503 TI - Introduction to a Virtual Special Issue on plant respiration in variable environments. PMID- 21631504 TI - Pushing back in time: the role of fire in plant evolution. PMID- 21631505 TI - The hairy problem of epigenetics in evolution. PMID- 21631506 TI - How do we improve Earth system models? Integrating Earth system models, ecosystem models, experiments and long-term data. 1st INTERFACE workshop, Captiva Island, FL, USA, 28 February-3 March 2011. PMID- 21631507 TI - Genes to ecosystems: exploring the frontiers of ecology with one of the smallest biological units. AB - Genes and their expression levels in individual species can structure whole communities and affect ecosystem processes. Although much has been written about community and ecosystem phenotypes with a few model systems, such as poplar and goldenrod, here we explore the potential application of a community genetics approach with systems involving invasive species, climate change and pollution. We argue that community genetics can reveal patterns and processes that otherwise might remain undetected. To further facilitate the community genetics or genes-to ecosystem concept, we propose four community genetics postulates that allow for the conclusion of a causal relationship between the gene and its effect on the ecosystem. Although most current studies do not satisfy these criteria completely, several come close and, in so doing, begin to provide a genetic-based understanding of communities and ecosystems, as well as a sound basis for conservation and management practices. PMID- 21631508 TI - An early treatment with 17-beta-estradiol is neuroprotective against the long term effects of neonatal ionizing radiation exposure. AB - Ionizing radiations can induce oxidative stress on target tissues, acting mainly through reactive oxygen species (ROS). The aim of this work was to investigate if 17-beta-estradiol (betaE) was able to prevent hippocampal-related behavioral and biochemical changes induced by neonatal ionizing radiation exposure and to elucidate a potential neuroprotective mechanism. Male Wistar rats were irradiated with 5 Gy of X-rays between 24 and 48 h after birth. A subset of rats was subcutaneously administered with successive injections of betaE or 17-alpha estradiol (alphaE), prior and after irradiation. Rats were subjected to different behavioral tasks to evaluate habituation and associative memory as well as anxiety levels. Hippocampal ROS levels and protein kinase C (PKC) activity were also assessed. Results show that although betaE was unable to prevent radiation induced hippocampal PKC activity changes, most behavioral abnormalities were reversed. Moreover, hippocampal ROS levels in betaE-treated irradiated rats approached control values. In addition, alphaE administered to irradiated animals was effective in preventing radiation-induced alterations. In conclusion, betaE was able to counteract behavioral and biochemical changes induced in irradiated animals, probably acting through an antioxidant mechanism. PMID- 21631509 TI - Effects of nonsurgical periodontal therapy on C-reactive protein and serum lipids in Jordanian adults with advanced periodontitis. AB - BACKGROUND AND OBJECTIVE: Data on whether periodontal therapy affects serum CRP levels are inconclusive. The aim of this study was to determine if nonsurgical periodontal therapy has any effect on CRP and serum lipid levels in patients with advanced periodontitis. MATERIAL AND METHODS: Thirty-six systemically healthy patients, >= 40 years of age and with advanced periodontitis, were recruited for the study. Patients were randomized consecutively to one of two groups: the treatment group (n = 18) or the control group (n = 18). Treated subjects received nonsurgical periodontal therapy, which included oral hygiene instructions and subgingival scaling and root planing. Systemic levels of inflammatory markers [C reactive protein (CRP) and the lipid profile] were measured at baseline and 3 mo after periodontal therapy. RESULTS: Nonsurgical periodontal therapy in the treatment group resulted in a significant reduction in the serum CRP level. The average CRP level decreased from 2.3 mg/dL at baseline to 1.8 mg/dL (p < 0.005) after 3 mo of periodontal therapy. The average reduction (95% confidence interval) in CRP was 0.498 (95% confidence interval = 0.265-0.731). In the treatment group, the reduction in CRP was significantly, linearly and directly correlated with the reduction in the plaque index, the gingival index and the percentage of sites with pocket depth >= 7 mm (Pearson correlation coefficient = 0.746, 0.425 and 0.621, respectively). Nonsurgical periodontal therapy had no effect on the lipid parameters. CONCLUSION: This study demonstrated that nonsurgical periodontal therapy results in a significant reduction in the serum CRP level. The effect of this outcome on systemic disease is still unknown. PMID- 21631510 TI - EMD or subepithelial connective tissue graft for the treatment of single gingival recessions: a pilot study. AB - BACKGROUND AND OBJECTIVE: The combination of EMD with root-coverage procedures has been shown to promote periodontal regeneration on the exposed root surface. The aim of this randomized clinical study was to compare the efficacy of EMD with connective tissue graft (CTG) for the treatment of Miller Class I and Class II single recessions in a split-mouth design. MATERIAL AND METHODS: The study included 12 systemically healthy patients, each with two single bilateral gingival recession defects (24 recessions). One recession defect in each patient was treated with the coronally advanced flap (CAF) + EMD procedure and the other recession defect was treated with the CTG + EMD procedure, in a split-mouth study design. The parameters recession depth (RD), recession width, percentage of root coverage, height of keratinized tissue (HKT), gingival thickness, probing depth and clinical attachment level (CAL) were recorded at baseline. All parameters, except for gingival thickness, were remeasured at 6 and 12 mo. RESULTS: The mean percentage root coverage at the final evaluation was 92 +/- 14% for the CAF + EMD group and 89 +/- 14% for the CAF + CTG group. Both treatments resulted in statistically significant (p < 0.05) decreases in RD and recession width, and increases in HKT, at 6 and 12 mo. There was also a significant decrease in the probing depth and a significant gain in the CAL for both groups. The probing depth was statistically higher in the CAF + CTG group than in the CAF + EMD group at 6 mo (p < 0.05), while the CAL was statistically lower in the CAF + EMD group than in the CAF + CTG group at 6 and 12 mo (p < 0.05). CONCLUSION: The present study demonstrated that both CAF + EMD and CAF + CTG procedures were similarly successful in treating Miller Class I and Class II single gingival recession defects. PMID- 21631511 TI - Cholesterol-lowering effects of oat beta-glucan. AB - Elevated total and low-density lipoprotein (LDL) cholesterol levels are considered major risk factors for cardiovascular disease. Oat beta-glucan, a soluble dietary fiber that is found in the endosperm cell walls of oats, has generated considerable interest due to its cholesterol-lowering properties. The United States Food and Drug Administration (FDA) approved a health claim for beta glucan soluble fiber from oats for reducing plasma cholesterol levels and risk of heart disease in 1997. Similarly, in 2004 the United Kingdom Joint Health Claims Initiative (JHCI) allowed a cholesterol-lowering health claim for oat beta glucan. The present review aims to investigate if results from more recent studies are consistent with the original conclusions reached by the FDA and JHCI. Results of this analysis show that studies conducted during the past 13 years support the suggestion that intake of oat beta-glucan at daily doses of at least 3 g may reduce plasma total and low-density lipoprotein (LDL) cholesterol levels by 5-10% in normocholesterolemic or hypercholesterolemic subjects. Studies described herein have shown that, on average, oat consumption is associated with 5% and 7% reductions in total and LDL cholesterol levels, respectively. Significant scientific agreement continues to support a relationship between oat beta-glucan and blood cholesterol levels, with newer data being consistent with earlier conclusions made by the FDA and JHCI. PMID- 21631512 TI - Inhibition of pattern recognition receptor-mediated inflammation by bioactive phytochemicals. AB - Emerging evidence reveals that pattern-recognition receptors (PRRs), Toll-like receptors (TLRs), and nucleotide-binding oligomerization domain proteins (NODs) mediate both infection-induced and sterile inflammation by recognizing pathogen associated molecular patterns and endogenous molecules, respectively. PRR mediated chronic inflammation is a determinant for the development and progression of chronic diseases including cancer, atherosclerosis, and insulin resistance. Recent studies demonstrated that certain phytochemicals inhibit PRR mediated pro-inflammation. Curcumin, helenalin, and cinnamaldehyde with alpha, beta-unsaturated carbonyl groups, or sulforaphane with an isothiocyanate group, inhibit TLR4 activation by interfering with cysteine residue-mediated receptor dimerization, while resveratrol, with no unsaturated carbonyl group, did not. Similarly, curcumin, parthenolide, and helenalin, but not resveratrol and (-) epigallocatechin-3-gallate (EGCG), also inhibit NOD2 activation by interfering with NOD2 dimerization. In contrast, resveratrol, EGCG, luteolin, and structural analogs of luteolin specifically inhibit TLR3 and TLR4 signaling by targeting TANK binding kinase 1 (TBK1) and receptor interacting protein 1 (RIP1) in Toll/IL 1 receptor domain-containing adaptor inducing IFN-beta (TRIF) complex. Together, these results suggest that PRRs and downstream signaling components are molecular targets for dietary strategies to reduce PRR-mediated chronic inflammation and consequent risks of chronic diseases. PMID- 21631513 TI - Evolution of food provision to athletes at the summer Olympic Games. AB - The history of food provision at the summer Olympic Games (OG) over the past century (1896-2008) provides insight into the evolution of sports nutrition research and the dietary strategies of athletes. Early research favoring protein as the main fuel for exercise was reflected in OG menus from 1932 to 1968. Despite conclusive research from the 1960s demonstrating the clear benefit of carbohydrate on exercise performance, a specific emphasis on carbohydrate-rich foods was not noted until the 1970s. Athlete food preferences and catering complexity evolved rapidly between 1970 and 2000, driven predominantly by a dramatic expansion of the OG and the emergence of systematic sports nutrition research. Nutritional advice by experts and sponsorship by food companies became increasingly important beginning with the 1984 Los Angeles OG. More recent developments include nutritional labeling of menu items and provision of a nutrition information desk (Barcelona 1992), demand for a "high-starch, low-fat menu" (Atlanta 1996), the addition of a dedicated menu website and the systematic gathering of information on athletes' apparent consumption (Sydney 2000), and appointment of the first international dietetic review committee (Beijing 2008). The history of catering at the OG tracks the evolution of sports nutrition practice from anecdotes and myth towards an established specialty in nutrition and dietetics grounded in evidence-based science. PMID- 21631514 TI - Calcium and obesity: effect size and clinical relevance. PMID- 21631515 TI - Efficacy of calcium supplementation for management of overweight and obesity: systematic review of randomized clinical trials. AB - Numerous dietary supplements are marketed as slimming aids, but the efficacy of most has not been proven. One such slimming aid is calcium. Presented here are the results of a systematic review that aimed to evaluate the evidence for or against the efficacy of calcium supplements for body-weight reduction in overweight and obese individuals. Electronic searches were conducted to identify relevant randomized clinical trials of at least 6 months duration. No restrictions of age, gender, language, or time of publication were imposed. Two reviewers independently determined the eligibility of studies, assessed the reporting quality of the studies included, and extracted data. Twenty-four eligible trials were identified, and seven were included. Five of the randomized clinical trials included were not of good reporting quality. A meta-analysis revealed a small, significant reduction in body weight for calcium compared with placebo (mean difference, (-) 0.74 kg; 95% confidence interval, (-) 1.00-(-) 0.48). A small, significant reduction in body fat favoring calcium over placebo was also noted (mean difference, (-) 0.93 kg; 95% confidence interval, (-) 1.16-( ) 0.71). In conclusion, the evidence from randomized clinical trials suggests calcium supplementation generates small, statistically significant weight loss in overweight and obese individuals, but the clinical relevance of this finding is uncertain. PMID- 21631517 TI - Serotonin transporter promoter region (5-HTTLPR) polymorphism predicts resting respiratory sinus arrhythmia. AB - Respiratory sinus arrhythmia (RSA) is often conceptualized as an index of physiological flexibility that has been related to emotion regulatory capacity. Although behavioral genetics research indicates that RSA is partly heritable, relatively few molecular genetics studies have been conducted. We examined whether the serotonin transporter promoter region (5-HTTLPR) polymorphism was associated with resting RSA among healthy young adults (N=71). Short 5-HTTLPR allele carriers had significantly lower resting RSA than long 5-HTTLPR homozygotes. Genotype explained 5% of the variance in resting RSA. Although firm conclusions depend on further study, the short allele of the 5-HTTLPR polymorphism may contribute to individual differences in RSA and its behavioral correlates. PMID- 21631518 TI - Approach and withdrawal actions modulate the startle reflex independent of affective valence and muscular effort. AB - The startle reflex is modulated during processing of pleasant and unpleasant affective cues. One explanation of this finding contends that approach and withdrawal motivational processes are key to explaining the effect. Undergraduates performed arm flexion and arm extension actions shown elsewhere to reliably elicit approach and withdrawal motives, respectively. Results showed that arm extension (a withdrawal action) was associated with the largest startles, followed by a neutral control action and arm flexion (an approach action). This pattern was not attributable to the subjective pleasantness or muscular effort associated with the actions. Results support motivational priming accounts of startle reflex modulation. PMID- 21631519 TI - The risk for cross-reactions after a cutaneous delayed-type hypersensitivity reaction to heparin preparations is independent of their molecular weight: a systematic review. AB - BACKGROUND: Heparins are a widely used class of drugs known to cause delayed-type hypersensitivity (DTH) reactions. Recent publications indicate that the incidence of these may be higher than previously thought. To date, patient-related but no drug-related risk factors for the development of DTH reactions to heparins have been identified, although molecular weight is discussed as a potentially relevant parameter. OBJECTIVES: To address this, a systematic review was conducted on the frequency of cross-reactions after DTH reactions to heparin preparations. METHODS: We electronically searched MEDLINE and EMBASE, hand-searched selected journals and references, and contacted experts for unpublished data. RESULTS: Sixty-six publications and unpublished data of 14 patients resulted in 198 patients with 1084 tests for cross-reactivity. The primary causative agents were mostly unfractionated heparin (50%) and low molecular weight heparins (49.5%). Cross-reactions were more likely after an initial DTH reaction to unfractionated heparin than after an initial DTH reaction to low molecular weight heparin. Our findings also indicate that molecular weight does not correlate with the risk for cross-reactivity, which is in line with recent observations, indicating that different heparins have to be individually considered. CONCLUSIONS: The available data demonstrated the lowest overall risk for cross-reactions for pentosan polysulfate (36.4%) and fondaparinux (10.4%). In the clinical context, fondaparinux is recommended as the current best alternative when a DTH reaction occurs. PMID- 21631521 TI - The high prevalence of peanut sensitization in childhood is due to cross reactivity to pollen. PMID- 21631520 TI - Hypersensitivity to nonsteroidal anti-inflammatory drugs (NSAIDs) - classification, diagnosis and management: review of the EAACI/ENDA(#) and GA2LEN/HANNA*. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are responsible for 21-25% of reported adverse drug events which include immunological and nonimmunological hypersensitivity reactions. This study presents up-to-date information on pathomechanisms, clinical spectrum, diagnostic tools and management of hypersensitivity reactions to NSAIDs. Clinically, NSAID hypersensitivity is particularly manifested by bronchial asthma, rhinosinusitis, anaphylaxis or urticaria and variety of late cutaneous and organ-specific reactions. Diagnosis of hypersensitivity to a NSAID includes understanding of the underlying mechanism and is necessary for prevention and management. A stepwise approach to the diagnosis of hypersensitivity to NSAIDs is proposed, including clinical history, in vitro testing and/or provocation test with a culprit or alternative drug depending on the type of the reaction. The diagnostic process should result in providing the patient with written information both on forbidden and on alternative drugs. PMID- 21631522 TI - Factor XII gene missense mutation Thr328Lys in an Arab family with hereditary angioedema type III. PMID- 21631523 TI - Chronic palpable purpura mediated by Kiwi antigen Act c 1-induced immune complex vasculitis. PMID- 21631524 TI - Cow's milk allergy as a cause of anaphylaxis to systemic corticosteroids. PMID- 21631525 TI - SRD5A2 gene mutations and polymorphisms in Spanish 46,XY patients with a disorder of sex differentiation. AB - One hundred and forty-six index patients with 46,XY DSD in whom gonads were confirmed as testes were consecutively studied for a molecular diagnosis during the period 2002-2010. AR gene was analysed in all patients as the first candidate gene, yielding a mutation in 42.5% of cases and SRD5A2 gene was analysed as the second candidate gene, resulting in the characterization of 10 different mutations (p.Y91D, p.G115D, p.Q126R, p.R171S, p.Y188CfsX9, p.N193S, p.A207D, p.F219SfsX60, p.R227Q and p.R246W) in nine index patients (6.2% of the total number of 46,XY DSD patients). One of the mutations (p.Y188CfsX9) has never been reported. In addition, we genotyped SRD5A2 gene p.V89L and c.281+15T>C polymorphisms in 46,XY DSD and in 156 normal adult males and found that patients with SRD5A2 mutations or without a known molecular diagnosis presented a higher frequency of homozygous p.L89, homozygous TT and combined CCTT genotypes compared with controls. This result suggests that 46,XY DSD patient phenotypes may be influenced by SRD5A2 polymorphism genotypes. SRD5A2 gene mutations may not be as infrequent as previously considered in 46,XY DSD patients with variable degrees of external genitalia virilization at birth and normal T production and appears to be the second aetiology in our series. PMID- 21631526 TI - Expression and interdependencies of pluripotency factors LIN28, OCT3/4, NANOG and SOX2 in human testicular germ cells and tumours of the testis. AB - OCT3/4, NANOG, SOX2 and, most recently, LIN28 have been identified as key regulators of pluripotency in mammalian embryonic and induced stem cells, and are proven to be crucial for generation of the mouse germ-cell lineage. These factors are a hallmark of certain histological types of germ-cell tumours (GCTs). Here, we report novel information on the temporal and spatial expression pattern of LIN28 during normal human male germ-cell development as well as various types of GCTs. To investigate LIN28 expression, immunohistochemical analyses and quantitative proximity ligation assay-based TaqMan protein assays were applied on snap-frozen and formalin-fixed, paraffin-embedded samples as well as representative cell lines. LIN28 was found in primordial germ cells, gonocytes and pre-spermatogonia, in contrast to OCT3/4 and NANOG, which were found only in the first two stages. LIN28 was also found in all precursor lesions (carcinoma in situ and gonadoblastoma) of type II GCTs, as well as the invasive components seminoma and the non-seminomatous elements embryonal carcinoma and yolk sac tumour. Choriocarcinoma showed a heterogeneous pattern, while teratomas and spermatocytic seminomas (type III GCTs) were negative. This expression pattern suggests that LIN28 is associated with malignant behaviour of type II GCTs. Cell line experiments involving siRNA knockdown of LIN28, OCT3/4 and SOX2 showed that LIN28 plays a role in the maintenance of the undifferentiated state of both seminoma and embryonal carcinoma, closely linked to, and likely upstream of OCT3/4 and NANOG. In conclusion, LIN28 regulates the differentiation status of seminoma and embryonal carcinoma and is likely to play a related role in normal human germ-cell development. PMID- 21631528 TI - Relative importance of prenatal and postnatal androgen action in determining growth of the penis and anogenital distance in the rat before, during and after puberty. AB - Experimental animal studies show that measurement of anogenital distance (AGD) and/or penis length may provide lifelong 'read-outs' of foetal androgen exposure during the masculinization programming window (MPW). However, variation in postnatal androgen exposure may complicate interpretation of such measurements. This is important to clarify if such measurements are to be applied to humans. The present aim was to evaluate effects of prenatal and/or postnatal manipulation of androgen production/action on growth of AGD and the penis in rats. Pregnant rats were treated daily before (e13.5-e21.5) and after birth (postnatal days 1 15) with either vehicle, 500 mg/kg di(n-butyl) phthalate (DBP) or 100 mg/kg flutamide (postnatal only) in prenatal + postnatal treatment combinations (N = 6 treatment combinations); DBP impairs androgen production whereas flutamide impairs androgen action. Male offspring were killed on postnatal day 8 (prepuberty), 25 (early puberty) or 90 (adulthood) when AGD was measured, the penis dissected out and its weight and length measured; plasma testosterone and ventral prostate weight were measured at day 90 to assess endogenous androgen exposure. In controls, penis length, girth and AGD increased 2.2-, 5.3-and 5.9 fold respectively from day 8 to day 90. Significant inhibition of penis growth and final length and girth was induced by treatments that inhibited postnatal androgen action. Conversely, growth and ultimate (adult) AGD was inhibited by prenatal inhibition of androgen production whereas postnatal androgen inhibition had negligible effect. Nevertheless, AGD and penis length were highly correlated at every age (R(2) > 0.33; p < 0.0001). However, altered endogenous androgen exposure may confound interpretation of changes in adults exposed prenatally/postnatally to DBP/flutamide. We conclude that AGD provides a lifelong guide to prenatal androgen exposure (in the MPW) whereas penis size reflects both prenatal + postnatal androgen exposure. At the group treatment level, prepubertal measurement of either AGD or penis size accurately predicts their size in adulthood. PMID- 21631527 TI - Associations between testicular hormones at adolescence and attendance at chlorinated swimming pools during childhood. AB - The goal was to evaluate the associations between testicular hormones at adolescence and the exposure to chlorination by-products when attending chlorinated swimming pools. We obtained serum samples from 361 school male adolescents (aged 14-18years) who had visited swimming pools disinfected with chlorine or by copper-silver ionization. We analysed serum concentrations of inhibin B (two different assays), total and free testosterone, sex hormone binding globulin, luteinizing hormone (LH), follicle stimulating hormone (FSH) and dehydroepiandrosterone sulphate (DHEAS). There were strong inverse associations between serum levels of inhibin B (both assays) or of total testosterone, adjusted or unadjusted for gonadotropins and the time adolescents had spent in indoor chlorinated pools, especially during their childhood. Adolescents having attended indoor chlorinated pools for more than 250h before the age of 10years or for more than 125h before the age of 7years were about three times more likely to have an abnormally low serum inhibin B and/or total testosterone (<10th percentile) than their peers who never visited this type of pool during their childhood (odds ratio, 95% CI, 2.83, 1.06-7.52, p=0.04 and 3.67, 1.45-9.34, p=0.006, respectively). Such associations were not seen with free testosterone, LH, FSH and DHEAS or with the attendance of outdoor chlorinated pools or of the copper-silver pool. Swimming in indoor chlorinated pools during childhood is strongly associated with lower levels of serum inhibin B and total testosterone. The absorption of reprotoxic chlorination by-products across the highly permeable scrotum might explain these associations. PMID- 21631529 TI - Stable extramarital affairs are breaking the heart. AB - The relationship between extramarital affairs and cardiovascular risk is still not completely clarified. The aim of this study was to investigate whether extramarital affairs have a protective effect on cardiovascular risk or, conversely, a deleterious one. Among patients studied, 91.8% of the whole sample reported no or occasional extramarital affairs, while 8.2% declared a stable secondary relationship. During a median follow-up of 4 [0-8] years, 95 major adverse cardiovascular events (MACE), eight of which were fatal, were observed. Cox analysis, after adjustment for confounding factors, showed that presence of stable extramarital affair was associated with a higher incidence of MACE (HR = 2.13 [1.12; 4.07], p = 0.023). The introduction in the Cox model of patient perceived partner's hypoactive sexual desire (PPPHSD) attenuates the association (HR 1.86 [0.93; 3.70], p = 0.078). The sample was therefore divided according to PPPHSD. We observed that unadjusted incidence of MACE was significantly associated with presence of extramarital affairs only in men reporting a primal partner without PPPHSD. This association was also confirmed in a Cox regression model, after adjusting for confounders (HR = 2.87 [1.81; 6.98], p = 0.020). We can conclude that to be unfaithful represents an independent risk factor for MACE. Therefore, infidelity induces not only heart trouble in the betrayed partners, but seems to be also able to increase the betrayer's heart-related events. PMID- 21631530 TI - EIN2 regulates salt stress response and interacts with a MA3 domain-containing protein ECIP1 in Arabidopsis. AB - Ethylene signalling regulates plant growth and development. However, its roles in salt stress response are less known. Here we studied functions of EIN2, a central membrane protein of ethylene signalling, and its interacting protein ECIP1 in salt stress responses. Mutation of EIN2 led to extreme salt sensitivity as revealed by phenotypic and physiological changes, and overexpression of C terminus of EIN2 suppressed salt sensitivity in ein2-5, indicating that EIN2 is required for salt tolerance. Downstream components EIN3 and EIL1 are also essential for salt tolerance because ein3-1eil1-1 double mutant showed extreme salt-sensitive phenotype. A MA3 domain-containing protein ECIP1 was further identified to interact with EIN2 in yeast two-hybrid assay and GST pull-down assay. Loss-of-function of ECIP1 resulted in enhanced ethylene response but altered salt response during seed germination and plant growth. Double mutant analysis revealed that ein2-1 was epistatic to ecip1, and ecip1 mutation partially suppressed ethylene-insensitivity of etr2-1 and ein4-1. These studies strengthen that interactions between ECIP1 and EIN2 or ethylene receptors regulate ethylene response and stress response. PMID- 21631531 TI - Carbonic anhydrase and the molecular evolution of C4 photosynthesis. AB - C(4) photosynthesis, a biochemical CO(2)-concentrating mechanism (CCM), evolved more than 60 times within the angiosperms from C(3) ancestors. The genus Flaveria, which contains species demonstrating C(3), C(3)-C(4), C(4)-like or C(4) photosynthesis, is a model for examining the molecular evolution of the C(4) pathway. Work with carbonic anhydrase (CA), and C(3) and C(4) Flaveria congeners has added significantly to the understanding of this process. The C(4) form of CA3, a beta-CA, which catalyses the first reaction in the C(4) pathway by hydrating atmospheric CO(2) to bicarbonate in the cytosol of mesophyll cells (mcs), evolved from a chloroplastic C(3) ancestor. The molecular modifications to the ancestral CA3 gene included the loss of the sequence encoding the chloroplast transit peptide, and mutations in regulatory regions that resulted in high levels of expression in the C(4) mesophyll. Analyses of the CA3 proteins and regulatory elements from Flaveria photosynthetic intermediates indicated C(4) biochemistry very likely evolved in a specific, stepwise manner in this genus. The details of the mechanisms involved in the molecular evolution of other C(4) plant beta-CAs are unknown; however, comparative genetics indicate gene duplication and neofunctionalization played significant roles as they did in Flaveria. PMID- 21631532 TI - Identification of a homolog of Arabidopsis DSP4 (SEX4) in chestnut: its induction and accumulation in stem amyloplasts during winter or in response to the cold. AB - Oligosaccharide synthesis is an important cryoprotection strategy used by woody plants during winter dormancy. At the onset of autumn, starch stored in the stem and buds is broken down in response to the shorter days and lower temperatures resulting in the buildup of oligosaccharides. Given that the enzyme DSP4 is necessary for diurnal starch degradation in Arabidopsis leaves, this study was designed to address the role of DSP4 in this seasonal process in Castanea sativa Mill. The expression pattern of the CsDSP4 gene in cells of the chestnut stem was found to parallel starch catabolism. In this organ, DSP4 protein levels started to rise at the start of autumn and elevated levels persisted until the onset of spring. In addition, exposure of chestnut plantlets to 4 degrees C induced the expression of the CsDSP4 gene. In dormant trees or cold-stressed plantlets, the CsDSP4 protein was immunolocalized both in the amyloplast stroma and nucleus of stem cells, whereas in the conditions of vegetative growth, immunofluorescence was only detected in the nucleus. The studies indicate a potential role for DSP4 in starch degradation and cold acclimation following low temperature exposure during activity-dormancy transition. PMID- 21631533 TI - Role of peroxidases in the compensation of cytosolic ascorbate peroxidase knockdown in rice plants under abiotic stress. AB - Current studies, particularly in Arabidopsis, have demonstrated that mutants deficient in cytosolic ascorbate peroxidases (APXs) are susceptible to the oxidative damage induced by abiotic stress. In contrast, we demonstrate here that rice mutants double silenced for cytosolic APXs (APx1/2s) up-regulated other peroxidases, making the mutants able to cope with abiotic stress, such as salt, heat, high light and methyl viologen, similar to non-transformed (NT) plants. The APx1/2s mutants exhibited an altered redox homeostasis, as indicated by increased levels of H2O2 and ascorbate and glutathione redox states. Both mutant and NT plants exhibited similar photosynthesis (CO2) assimilation and photochemical efficiency) under both normal and stress conditions. Overall, the antioxidative compensatory mechanism displayed by the mutants was associated with increased expression of OsGpx genes, which resulted in higher glutathione peroxidase (GPX) activity in the cytosolic and chloroplastic fractions. The transcript levels of OsCatA and OsCatB and the activities of catalase (CAT) and guaiacol peroxidase (GPOD; type III peroxidases) were also up-regulated. None of the six studied isoforms of OsApx were up-regulated under normal growth conditions. Therefore, the deficiency in cytosolic APXs was effectively compensated for by up-regulation of other peroxidases. We propose that signalling mechanisms triggered in rice mutants could be distinct from those proposed for Arabidopsis. PMID- 21631534 TI - Characterization of C3--C4 intermediate species in the genus Heliotropium L. (Boraginaceae): anatomy, ultrastructure and enzyme activity. AB - Photosynthetic pathway characteristics were studied in nine species of Heliotropium (sensu lato, including Euploca), using assessments of leaf anatomy and ultrastructure, activities of PEP carboxylase and C4 acid decarboxylases, and immunolocalization of ribulose 1.5-bisphosphate carboxylase/oxygenase (Rubisco) and the P-subunit of glycine decarboxylase (GDC). Heliotropium europaeum, Heliotropium calcicola and Heliotropium tenellum are C3 plants, while Heliotropium texanum and Heliotropium polyphyllum are C4 species. Heliotropium procumbens and Heliotropium karwinskyi are functionally C3, but exhibit 'proto Kranz' anatomy where bundle sheath (BS) cells are enlarged and mitochondria primarily occur along the centripetal (inner) wall of the BS cells; GDC is present throughout the leaf. Heliotropium convolvulaceum and Heliotropium greggii are C3--C4 intermediates, with Kranz-like enlargement of the BS cells, localization of mitochondria along the inner BS wall and a loss of GDC in the mesophyll (M) tissue. These C3--C4 species of Heliotropium probably shuttle photorespiratory glycine from the M to the BS tissue for decarboxylation. Heliotropium represents an important new model for studying C4 evolution. Where existing models such as Flaveria emphasize diversification of C3--C4 intermediates, Heliotropium has numerous C3 species expressing proto-Kranz traits that could represent a critical initial phase in the evolutionary origin of C4 photosynthesis. PMID- 21631535 TI - Day length is a key regulator of transcriptomic responses to both CO(2) and H(2)O(2) in Arabidopsis. AB - Growth day length, CO(2) levels and H(2)O(2) all impact plant function, but interactions between them remain unclear. Using a whole-genome transcriptomics approach, we identified gene expression patterns responding to these three factors in Arabidopsis Col-0 and the conditional catalase-deficient mutant, cat2. Plants grown for 5 weeks at high CO(2) in short days (hCO(2)) were transferred to air in short days (SD air) or long days (LD air), and microarray data produced were subjected to three independent studies. The first two analysed genotype independent responses. They identified 1549 genes differentially expressed after transfer from hCO(2) to SD air. Almost half of these, including genes modulated by sugars or associated with redox, stress or abscisic acid (ABA) functions, as well as light signalling and clock genes, were no longer significant after transfer to air in LD. In a third study, day length-dependent H(2)O(2)-responsive genes were identified by comparing the two genotypes. Two clearly independent responses were observed in cat2 transferred to air in SD and LD. Most H(2)O(2) responsive genes were up-regulated more strongly in SD air. Overall, the analysis shows that both CO(2) and H(2)O(2) interact with day length and photoreceptor pathways, indicating close networking between carbon status, light and redox state in environmental responses. PMID- 21631536 TI - The influence of ascorbate on anthocyanin accumulation during high light acclimation in Arabidopsis thaliana: further evidence for redox control of anthocyanin synthesis. AB - Ascorbate and anthocyanins act as photoprotectants during exposure to high light (HL). They accumulate in Arabidopsis leaves in response to HL on a similar timescale, suggesting a potential relationship between them. Flavonoids and related metabolites were identified and profiled by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The ascorbate-deficient mutants vtc1, vtc2 and vtc3 accumulated less anthocyanin than wild-type (WT) during HL acclimation. In contrast, kaempferol glycoside accumulation was less affected by light and not decreased by ascorbate deficiency, while sinapoyl malate levels decreased during HL acclimation. Comparison of six Arabidopsis ecotypes showed a positive correlation between ascorbate and anthocyanin accumulation in HL. mRNA-Seq analysis showed that all flavonoid biosynthesis transcripts were increased by HL acclimation in WT. RT-PCR analysis showed that vtc1 and vtc2 were impaired in HL induction of transcripts of anthocyanin biosynthesis enzymes, and the transcription factors PAP1, GL3 and EGL3 that activate the pathway. Abscisic acid (ABA) and jasmonic acid (JA), hormones that could affect anthocyanin accumulation, were unaffected in vtc mutants. It is concluded that HL induction of anthocyanin synthesis involves a redox-sensitive process upstream of the known transcription factors. Because anthocyanins accumulate in preference to kaempferol glycosides and sinapoyl malate in HL, they might have specific properties that make them useful in HL acclimation. PMID- 21631537 TI - The low temperature response pathways for cold acclimation and vernalization are independent. AB - Vernalization is the promotion of flowering in response to the prolonged cold of winter. To survive sub-zero winter temperatures, plants must first acclimate to low, non-freezing temperatures (cold acclimation). Induction of VERNALIZATION INSENSITIVE 3 (VIN3), the first gene in the vernalization pathway, is initiated within the same time frame as the induction of genes in the cold acclimation pathway raising the question of whether there are common elements in the signal transduction pathways that activate these two responses to cold. We show that none of the signalling components required for cold acclimation, including the 'master regulator'INDUCTION OF CBF EXPRESSION1 (ICE1) or HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENE1 (HOS1), which has been described as a link between cold acclimation and vernalization, play a role in VIN3 induction. We also show that the hormone abscisic acid (ABA) does not modulate VIN3 induction, consistent with earlier reports that ABA signalling plays no role in the vernalization response. The cold acclimation pathway is activated at 12 degrees C, at which temperature there is no induction of VIN3 expression. Taken together, our data demonstrate that the responses to low temperatures leading to cold acclimation and vernalization are controlled by distinct signalling pathways. PMID- 21631538 TI - Links between root developmental traits and foraging performance. AB - We designed a simple dynamic and stochastic architectural model with six parameters to link the foraging performance of root systems to their developmental processes. Soil foraging was quantified by the volume enveloping the roots until a given uptake distance. Many simulated architectures were obtained by combining four different values for each parameter. The rate of soil colonization was mainly defined by individual root elongation rates and interbranch distances. Less intuitively, we showed that differentiation of elongation rates among the roots increased this colonization rate. Uptake efficiency--the ratio of the actual colonized volume to the volume of a unique cylinder with the same length and a radius corresponding to the uptake distance- declined with root system size. Nevertheless, large variations in efficiency existed among root systems for a given size, typically in a 4- to 10-fold range. Therefore, the 'efficiency gain' was defined as the deviation from the average trend in efficiency versus size. Between-root differentiation in elongation rates increased this gain. The level of hierarchy between mother and lateral roots, as well as the variation of elongation rates among lateral roots, was also shown to contribute to this optimization. Several parameter combinations could lead to similar efficiency gains. PMID- 21631539 TI - On reducing an empathy gap: the impact of self-construal and order of judgment. AB - Empathy gaps, in which individuals exaggerate self-other similarities or differences, generate errors in social judgments. We investigated whether changing individuals' self-construal may reduce one specific empathy gap: the illusion of courage. Participants primed with independent or interdependent self construal made judgments about their own and other people's willingness to dance in public. Participants in the interdependence condition showed a reduction of the empathy gap, but only when judging the other first. This finding highlights that simple contextual manipulations have the potential to reduce egocentric biases in social judgments. PMID- 21631540 TI - Persistent escalation of alcohol drinking in C57BL/6J mice with intermittent access to 20% ethanol. AB - BACKGROUND: Intermittent access (IA) to drugs of abuse, as opposed to continuous access, is hypothesized to induce a kindling-type transition from moderate to escalated use, leading to dependence. Intermittent 24-hour cycles of ethanol access and deprivation can generate high levels of voluntary ethanol drinking in rats. METHODS: The current study uses C57BL/6J mice (B6) in an IA to 20% ethanol protocol to escalate ethanol drinking levels. Adult male and female B6 mice were given IA to 20% ethanol on alternating days of the week with water available ad libitum. Ethanol consumption during the initial 2 hours of access was compared with a short-term, limited access "binge" drinking procedure, similar to drinking in-the-dark (DID). B6 mice were also assessed for ethanol dependence with handling-induced convulsion, a reliable measure of withdrawal severity. RESULTS: After 3 weeks, male mice given IA to ethanol achieved high stable levels of ethanol drinking in excess of 20 g/kg/24 h, reaching above 100 mg/dl blood ethanol concentrations, and showed a significantly higher ethanol preference than mice given continuous access to ethanol. Also, mice given IA drank about twice as much as DID mice in the initial 2-hour access period. B6 mice that underwent the IA protocol for longer periods of time displayed more severe signs of alcohol withdrawal. Additionally, female B6 mice were given IA to ethanol and drank significantly more than males (ca. 30 g/kg/24 h). DISCUSSION: The IA method in B6 mice is advantageous because it induces escalated, voluntary, and preferential per os ethanol intake, behavior that may mimic a cardinal feature of human alcohol dependence, though the exact nature and site of ethanol acting in the brain and blood as a result of IA has yet to be determined. PMID- 21631541 TI - Functioning of alcohol use disorder criteria among men and women with arrests for driving under the influence of alcohol. AB - BACKGROUND: Many states require screening of individuals arrested for driving under the influence (DUI) of alcohol to determine recidivism risk and the need for treatment based on severity of alcohol problems. Several screening instruments use DSM-IV criteria for alcohol abuse and dependence to assess alcohol problems in this population, but whether they adequately measure alcohol problems in individuals with DUIs has not been examined. In addition, gender differences in DUI samples suggest that female offenders have more severe alcohol problems than male offenders. The current study examines differences in alcohol criteria functioning by DUI history and gender using an item response theory (IRT) approach. METHODS: Data from diagnostic interviews with 8,605 participants in the Collaborative Study on the Genetics of Alcoholism, including 1,655 who ever reported a DUI arrest (20% women), were used to examine differences in alcohol criteria functioning between men and women with and without DUIs. The factor underlying item response was conceptualized as unidimensional, representing alcohol problem severity. RESULTS: Social/interpersonal problems, larger/longer, and inability/persistent desire to quit displayed greater discrimination of IRT-defined alcohol problem severity among individuals with DUIs than those without. Irrespective of DUI status, women had a higher threshold than men for time spent drinking or recovering. Women without DUIs had a higher threshold than similar men for social/interpersonal problems. Taken as a whole, the criteria yielded similar amounts of information in all groups. CONCLUSIONS: DSM-IV criteria for alcohol abuse and dependence adequately detect alcohol problem severity in individuals with DUIs, and some are better at detecting severity in this particularly high-risk group than in individuals without DUIs. However, the criteria as a whole are equally effective in measuring alcohol problem severity among individuals with and without DUIs and may be used with confidence in screening DUI offenders. PMID- 21631542 TI - Neurocognitive performance, alcohol withdrawal, and effects of a combination of flumazenil and gabapentin in alcohol dependence. AB - BACKGROUND: Among some alcohol-dependent individuals, early alcohol abstinence is marked by alcohol withdrawal (AW), a phenomenon mediated by GABA and glutamate signaling. We previously reported that a combination of 2 medications that affect GABA and glutamate tone, gabapentin and flumazenil, more effectively reduced drinking among individuals with higher pretreatment AW (Anton et al., 2009). This study evaluated whether this finding is related to changes in neurocognitive performance, which is also affected by cortical GABA and glutamate tone. METHODS: Neurocognitive performance was assessed at baseline and twice during the first week of treatment among 60 alcohol-dependent participants in the previously published clinical trial. RESULTS: AW was associated with poorer baseline performance on 4 of 8 measures, and individuals with higher baseline AW who received the gabapentin and flumazenil combination demonstrated greater improvement on a measure of response inhibition than those with lower AW or those who received a combination of placebos. Improvement in response inhibition during the first week and medication group interacted in their effect on subsequent drinking, such that improvement predicted greater abstinence only among individuals who received gabapentin and flumazenil. Improvement on other neurocognitive measures was neither differentially impacted by medication or baseline AW nor related to subsequent drinking. CONCLUSIONS: Taken together, these data suggest that acute AW accounts for a small proportion of variance in neurocognitive performance, that gabapentin and flumazenil slightly improve response inhibition during early abstinence, and that such improvement may somewhat reduce later drinking. However, these medications may not affect other neurocognitive domains. PMID- 21631543 TI - The effects of short-term chronic ethanol intoxication and ethanol withdrawal on the molecular composition of the rat hippocampus by FT-IR spectroscopy. AB - BACKGROUND: The numerous adverse effects of ethanol abuse and ethanol withdrawal on biological systems are well documented. Conversely, the understanding of the molecular mechanisms underlying these pathological effects is still incomplete. This study was undertaken to investigate the effects of short-term chronic ethanol administration and ethanol withdrawal on the molecular structure and function of hippocampal tissue, a brain region important for mnemonic processes and known to be highly susceptible to ethanol intoxication. METHODS: Ethanol was administered to adult Wistar rats by intragastric intubation for 15 days with a stepwise increase in the daily dose from 6 to 12 g/kg body weight, with the highest dose delivered for the last 2 days only. The total daily dose of ethanol was divided into 3 equal portions administered 4 hours apart. Animals were sacrificed by decapitation at 4, 24, and 72 hours after the last ethanol administration to examine potential effects of ethanol intoxication and ethanol withdrawal. Ethanol-related molecular changes were monitored by Fourier transform infrared (FT-IR) spectroscopy. RESULTS: Significant changes in the hippocampal content, structure, and function of lipids, proteins, and nucleic acids were recorded under ethanol intoxication. Seventy-two hours after the cessation of ethanol administration, during the late phase of withdrawal, alterations in the macromolecules' content and conformational changes in protein and nucleic acid structure ameliorated, while the changes in macromolecular ratios, lipid order, and dynamics aggravated. CONCLUSIONS: Our results suggest that 15 days of binge like drinking resulting in the high blood alcohol concentration (varying in the dose-dependent manner between 253 and 606 mg/dl) produced a strong physical dependence manifested mainly by the changes in lipid profiles pointing toward withdrawal-induced oxidative stress. These results show that ethanol withdrawal may cause equal to or even more severe brain damage than the ethanol itself, which should be considered when designing antialcohol therapies. PMID- 21631544 TI - Impulsivity is an independent predictor of 15-year mortality risk among individuals seeking help for alcohol-related problems. AB - BACKGROUND: Although past research has found impulsivity to be a significant predictor of mortality, no studies have tested this association in samples of individuals with alcohol-related problems or examined moderation of this effect via socio-contextual processes. The current study addressed these issues in a mixed-gender sample of individuals seeking help for alcohol-related problems. METHODS: Using Cox proportional hazard models, variables measured at baseline and Year 1 of a 16-year prospective study were used to predict the probability of death from Years 1 to 16 (i.e., 15-year mortality risk). There were 628 participants at baseline (47.1% women); 515 and 405 participated in the follow-up assessments at Years 1 and 16, respectively. Among Year 1 participants, 93 individuals were known to have died between Years 1 and 16. RESULTS: After controlling for age, gender, and marital status, higher impulsivity at baseline was associated with an increased risk of mortality from Years 1 to 16; however, this association was accounted for by the severity of alcohol use at baseline. In contrast, higher impulsivity at Year 1 was associated with an increased risk of mortality from Years 1 to 16, and remained significant when accounting for the severity of alcohol use, as well as physical health problems, emotional discharge coping, and interpersonal stress and support at Year 1. In addition, the association between Year 1 impulsivity and 15-year mortality risk was moderated by interpersonal support at Year 1, such that individuals high on impulsivity had a lower mortality risk when peer/friend support was high than when it was low. CONCLUSIONS: The findings highlight impulsivity as a robust and independent predictor of mortality and suggest the need to consider interactions between personality traits and socio-contextual processes in the prediction of health related outcomes for individuals with alcohol use disorders. PMID- 21631545 TI - Soil-specific limitations for access and analysis of soil microbial communities by metagenomics. AB - Metagenomics approaches represent an important way to acquire information on the microbial communities present in complex environments like soil. However, to what extent do these approaches provide us with a true picture of soil microbial diversity? Soil is a challenging environment to work with. Its physicochemical properties affect microbial distributions inside the soil matrix, metagenome extraction and its subsequent analyses. To better understand the bias inherent to soil metagenome 'processing', we focus on soil physicochemical properties and their effects on the perceived bacterial distribution. In the light of this information, each step of soil metagenome processing is then discussed, with an emphasis on strategies for optimal soil sampling. Then, the interaction of cells and DNA with the soil matrix and the consequences for microbial DNA extraction are examined. Soil DNA extraction methods are compared and the veracity of the microbial profiles obtained is discussed. Finally, soil metagenomic sequence analysis and exploitation methods are reviewed. PMID- 21631546 TI - Surfactants expressed by soil pseudomonads alter local soil-water distribution, suggesting a hydrological role for these compounds. AB - Several biological roles have been demonstrated for surfactants expressed by soil and rhizosphere Pseudomonas spp., but the impact of these powerful surface-active agents on the local soil-water distribution within the partially saturated soil pore network has not been examined. To investigate this potential hydrological role, the liquid surface tension (gamma)-reducing activities (LSTRA) of 72 pseudomonads isolated from a sandy loam soil by tensiometry of culture supernatants were characterized. Of these, 67% exhibited LSTRA, reducing gamma to a minimum (gamma(Min)) of 24 mN m(-1) established by individual distribution identification analysis. Soil microcosms were then used to examine the impact of surfactant expression on the local soil-water distribution. The volumetric water content (theta) of soil microcosms was significantly lowered (0.78 *) by Pseudomonas fluorescens SBW25 expressing the surfactant viscosin compared with a surfactant-deficient mutant (P<0.002). Six of 15 soil pseudomonad isolates examined were found to have a similar impact on theta when compared with sterile microcosms (P<0.05). These findings indicate that surfactant-expressing pseudomonads could modify the local soil-water distributions and that surfactants may therefore play a significant hydrological role in soils, in addition to their recognized biological activities. PMID- 21631547 TI - The use of multiple typing methods allows a more accurate molecular characterization of Vibrio parahaemolyticus strains isolated from the Italian Adriatic Sea. AB - Vibrio parahaemolyticus is a natural inhabitant of marine environments and constitutes part of the autochthonous microbial communities, but is also associated with human gastroenteritis, wound infections and septicemia. Recently, a number of clinical cases of infection due to ingestion of seafood contaminated with V. parahaemolyticus and potentially pandemic marine strains isolated from water and plankton have been reported in Europe. To identify the source of virulent strains and to analyze the possible persistence, in time and space, of particular clones, the molecular typing of Vibrio strains is of high epidemiological interest. In this study, we applied pulsed-field gel electrophoresis and two PCR-based techniques (enterobacterial repetitive intergenic consensus- and repetitive extragenic palindromic-PCR) to establish the DNA fingerprints for the analysis of genetic variability among the environmental V. parahaemolyticus strains isolated in the area of the Venetian Lagoon. A temporal distribution of the environmental strains in the studied geographical area and, in some cases, a strong association between a certain genetic profile and a specific source have been evidenced. A number of genetic clusters/clones seem to persist over time, reappearing in the marine environment for subsequent months and also at a 1-year gap. The use of multiple typing methods allowed a more accurate characterization of the environmental strain genetic profiles and the identification of clones hardly revealed through common techniques. PMID- 21631548 TI - High expression levels of chitinase genes in Streptomyces coelicolor A3(2) grown in soil. AB - Although Streptomyces species are major chitin-degraders in soil ecosystems, the expression of the diverse chitinase genes within Streptomyces coelicolor grown in soil has not been assessed. As a first step, the induction pattern of nine chitinase genes in S. coelicolor growing in autoclaved soil was compared with those in liquid cultures. The relative expression levels of nine chitinase genes were measured using real-time reverse transcription PCR. The expression of all chitinase genes was induced by chitin in both autoclaved soil and liquid cultures, but to different levels. The expression levels of five chitinase genes in autoclaved soil were significantly higher than those in the liquid cultures. In particular, a putative chitinase gene, chitinase H, showed the highest induction in autoclaved soil. The same induction pattern was confirmed in nonautoclaved soil, indicating that soil contains some factors affecting the expression of chitinase genes. The chiH gene product, ChiH, cloned in Streptomycetes lividans was secreted and exhibited chitin degradation activity that was stable within a wide range of acidic pHs. The disruption of dasR, a transcriptional regulator for the uptake of N-acetylglucosamine, abolished the expression of chiH, demonstrating that DasR is required for the regulation of ChiH expression. PMID- 21631549 TI - Patterns of lignin degradation and oxidative enzyme secretion by different wood- and litter-colonizing basidiomycetes and ascomycetes grown on beech-wood. AB - The degradation of lignocellulose and the secretion of extracellular oxidoreductases were investigated in beech-wood (Fagus sylvatica) microcosms using 11 representative fungi of four different ecophysiological and taxonomic groups causing: (1) classic white rot of wood (e.g. Phlebia radiata), (2) 'nonspecific' wood rot (e.g. Agrocybe aegerita), (3) white rot of leaf litter (Stropharia rugosoannulata) or (4) soft rot of wood (e.g. Xylaria polymorpha). All strong white rotters produced manganese-oxidizing peroxidases as the key enzymes of ligninolysis (75-2200 mU g(-1)), whereas lignin peroxidase activity was not detectable in the wood extracts. Interestingly, activities of two recently discovered peroxidases - aromatic peroxygenase and a manganese independent peroxidase of the DyP-type - were detected in the culture extracts of A. aegerita (up to 125 mU g(-1)) and Auricularia auricula-judae (up to 400 mU g( 1)), respectively. The activity of classic peroxidases correlated to some extent with the removal of wood components (e.g. Klason lignin) and the release of small water-soluble fragments (0.5-1.0 kDa) characterized by aromatic constituents. In contrast, laccase activity correlated with the formation of high-molecular mass fragments (30-200 kDa). The differences observed in the degradation patterns allow to distinguish the rot types caused by basidiomycetes and ascomycetes and may be suitable for following the effects of oxidative key enzymes (ligninolytic peroxidases vs. laccases, role of novel peroxidases) during wood decay. PMID- 21631550 TI - Bifidobacterium bifidum MIMBb75 in irritable bowel syndrome. PMID- 21631552 TI - Limitations of retrospective IL28B polymorphisms and IP-10 real life studies in hepatitis C. PMID- 21631553 TI - Disruption of sleep architecture in minimal hepatic encephalopathy and ghrelin secretion. PMID- 21631554 TI - Single-drug treatments for chronic hepatitis B: summarising current information by network meta-analysis. PMID- 21631555 TI - Early normalisation of aminotransferase predicts complete biochemical remission in autoimmune hepatitis patients. PMID- 21631556 TI - Eosinophilic oesophagitis in noncardiac chest pain. PMID- 21631558 TI - Abnormal liver tests in people aged 75 and above: prevalence and association with mortality. AB - BACKGROUND: Despite their common use the occurrence and consequences of abnormal liver tests remain unclear. AIMS: To estimate the prevalence and mortality associated with abnormal liver tests in people aged 75 years and above. METHODS: A cohort study on 13,276 people aged 75 years and above, registered with general practices, with a valid measurement of one or more liver test, calculating the prevalence of abnormal aspartate transaminase (AST), alkaline phosphatase (ALP) or bilirubin. Hazard ratios (HRs) were calculated for all-cause and cause specific mortality comparing elderly patients with abnormal liver tests to elderly patients with normal liver tests. RESULTS: At least one abnormal liver test was found in 2175 subjects (16.1%, 95% CI [15.4%, 16.7%]). The prevalence of a single abnormal liver test was 3.3% (95% CI [3.0%, 3.7%]) for AST, 9.2% (95% CI [8.8%, 9.7%]) for ALP and 5.4% (95% CI [5.1%, 5.9%]) for bilirubin. Abnormal AST, ALP and bilirubin were associated with increased risks of all-cause mortality; adjusted HRs, 1.27(95% CI [1.09, 1.47]), 1.47(95% CI [1.35, 1.61]) and 1.15(95% CI [1.02, 1.30]), respectively. Abnormal AST and ALP were associated with sevenfold and sixfold increased risk of death from liver disease, respectively. Two or more abnormal liver tests were associated with 2-fold and 17-fold increased risk of death from cancer and liver disease, respectively. Of the causes examined, absolute mortality rates were highest for cardiovascular disease in subjects with and without abnormal liver tests. CONCLUSIONS: Abnormal liver tests occur commonly in elderly people and are associated with a modest increase in all-cause mortality. There was a strong association with liver disease; however, the majority of deaths were not due to this cause. PMID- 21631559 TI - Dual cut-off transient elastography to assess liver fibrosis in chronic hepatitis B: a cohort study with internal validation. AB - BACKGROUND: Transient elastography has gained popularity to stage liver fibrosis in chronic viral hepatitis, however, diagnostic cut-offs for severe fibrosis in chronic hepatitis B are poorly defined. AIM: To evaluate an algorithm with two distinct cut-offs for positive and negative prediction of significant fibrosis and cirrhosis in chronic hepatitis B patients. METHODS: Two cohorts of treatment naive patients with chronic hepatitis B (125 training and 92 validations) were consecutively and concurrently examined by percutaneous liver biopsy and transient elastography. Fibrosis was staged by Metavir (significant fibrosis = F >= 2; cirrhosis = F4) in >= 2 cm long liver tissue cores. RESULTS: A >13.1 kPa positive and a <= 9.4 kPa negative cut-off for cirrhosis had a >90% sensitivity and specificity, with an accuracy of 94%. The corresponding cut-offs for F >= 2 were >9.4 and <= 6.2 kPa, thus classifying 56% of patients with an overall accuracy of 90%. In the validation cohort, F4 and F >= 2 were predicted by the above transient elastography cut-offs with an overall accuracy >90%. In 165 patients with higher than upper limit of normal transaminase activity the dual cut-off algorithm of transient elastography was as accurate as in the 52 patients with normal alanine aminotransferase values in the prediction and exclusion of cirrhosis, only. CONCLUSIONS: A dual cut-off algorithm allowed for correctly classifying both significant fibrosis and cirrhosis in the majority of the patients with chronic hepatitis B, independent of alanine aminotransferase values, thus reducing the need for liver biopsy investigations. PMID- 21631560 TI - Association between the presence of brown adipose tissue and non-alcoholic fatty liver disease in adult humans. AB - BACKGROUND: The presence of active brown adipose tissue (BAT) has been associated with a reduced risk of obesity in adult humans. AIM: To examine whether the presence and activity of BAT in patients undergoing PET-CT examinations is related to the presence of fatty liver. METHOD: We retrospectively analysed 3666 consecutive PET-CT whole-body scans performed on a total of 1832 patients who were referred for suspected malignancies. BAT-positive subjects (BAT+) were defined as subjects who showed substantial amounts of brown adipose tissue on PET CT scans. In areas where uptake of [(18)F]FDG was identified by CT for BAT, the maximal standardised uptake values (SUVmax), defined as the maximum activity per millilitre within the region of interest divided by the injected dose in megabecquerels per gram of body weight, were determined. A ratio of mean liver attenuation to spleen attenuation <0.8 on CT scans was considered to indicate NAFLD. RESULTS: Thirty patients of the 1832 screened individuals (2%) demonstrated brown fat uptake (BAT+ subjects). Ninety matched individuals without evidence of BAT on PET scans (BAT- subjects) were enrolled for comparison purposes. After adjustment for potential confounders, the odds ratio for having NAFLD was significantly higher for BAT- subjects (3.12, 95% confidence interval = 1.03-9.88, P < 0.05). The SUVmax for brown fat tissue was significantly correlated with the ratio of mean liver attenuation to spleen attenuation (P < 0.05). CONCLUSION: The presence of brown adipose tissue in adulthood is independently associated with a lower likelihood of NAFLD diagnosed by CT findings. PMID- 21631561 TI - Veterinary clinical pathologists have an impact on science and the biopharmaceutical industry. PMID- 21631562 TI - Best practices for evaluation of bone marrow in nonclinical toxicity studies. AB - This manuscript is intended to provide a best practice approach to accurately and consistently assess toxicant-induced bone marrow effects of test articles. In nonclinical toxicity studies, complete blood count data in conjunction with the histological examination of the bone marrow are recommended as the foundation for assessing the effect of test articles on the hematopoietic system. This approach alone can be used successfully in many studies. However, in some situations it may be necessary to further characterize effects on the different hematopoietic lineages, either by cytological or flow cytometric evaluation of the bone marrow. Both modalities can be used successfully, and which one is selected will depend on the expertise, preference of the facility, and the nature of the change in the bone marrow. Other specialized techniques such as clonogenic assays or electron microscopy are used rarely to further characterize hematotoxicity. The indications and techniques to successfully employ histological, cytological, or flow cytometric evaluation as well as clonogenic assays and electron microscopy are reviewed. PMID- 21631563 TI - The elusive globule leukocyte. PMID- 21631565 TI - Paraneoplastic production of GM-CSF. PMID- 21631566 TI - Bison and copper. PMID- 21631567 TI - Development of a specific indicator of the well-being of wearers of removable dentures. AB - OBJECTIVES: Bearers of removable prostheses have certain particular characteristics that require the inclusion of items able to detect the specific effect on their oral functions (derived from the presence of such prostheses). We aimed to develop and validate a specific indicator for the assessment of oral health-related quality of life in bearers of dentures. METHODS: A panel of experts compiled a questionnaire addressing oral health-related quality of life (11 items) to assess the degree of well-being afforded by removable dentures. After a pilot trial in patients bearing prostheses (n = 36), a consecutive sampling of denture wearers (n = 123) was conducted among patients not requesting odontological treatment and accompanying persons, attending their respective health centres in the City of Granada and its metropolitan area. The subjects were classified as RPP1 (removable partial prosthesis on one of the arches, RPP2 (on both) and complete prosthesis (CP on both arches). RESULTS: The questionnaire was consistent (Cronbach alpha 0.814) and proved to have suitable validity. Factor analysis revealed three factors termed physical, psychological and social well-being, which coherently supported the interrelationship among the 11 items. The prevalence of impact was significantly higher in the CP individuals (97.1%) and in that of the RPP2 group (92.1%) than in the RPP1 subjects (80.7%). Prosthetic fit, aesthetics and self-confidence were significantly discrepant among the prosthetic groups, although the total prosthetic quality of life score was not able to differentiate them. The main prosthetic impact in all the prosthesis-wearing groups was attributed to 'food impaction', although all the groups were essentially satisfied with their mouth. CONCLUSIONS: The prosthetic quality of life (PQL) questionnaire is a valid and reliable indicator of the specific well-being of partial or complete denture wearers. PMID- 21631568 TI - Celecoxib does not alter cardiovascular and renal function during dietary salt loading. AB - 1. Cyclo-oxygenase-2 (COX-2)-derived prostaglandins are important in controlling sodium excretion and renin release. In the present study, we tested the hypothesis that a clinical dose of celecoxib would impair urinary sodium excretion and elevate blood pressure (BP) during dietary salt loading. 2. Twelve normotensive individuals (mean (+/- SEM) age 35 +/- 2 years) completed two separate 17 day dietary perturbations, one taking 200 mg/day celecoxib (CX2) and the other taking placebo (PL), randomized with a 1 month wash out. The controlled 17 day diet consisted of a 3 day run-in diet, 7 days of a low-salt (LS, 20 mmol sodium/day) diet and 7 days of a high-salt diet (HS, 350 mmol sodium/day) diet. The order in which the diets were applied was randomized. Data were collected on the last day of the LS and HS diets. 3. Plasma and urinary prostaglandins were modestly lower during celecoxib (P < 0.05). Urinary sodium excretion was greater (P < 0.01) during the HS diet (253 +/- 10 vs 281 +/- 27 mmol/24 h for PL vs CX2, respectively) compared with the LS diet (14 +/- 3 vs 17 +/- 7 mmol/24 h for PL vs CX2, respectively; P(drug) = 0.26). Celecoxib did not alter creatinine clearance (P > 0.50). Twenty-four hour mean arterial BP was similar during PL (87 +/- 2 vs 87 +/- 2 mmHg for LS and HS, respectively) and CX2 (88 +/- 2 vs 87 +/- 2 mmHg for LS and HS, respectively; P = 0.85), with no effect of dietary salt (P > 0.80). Plasma renin activity, angiotensin II and aldosterone were all suppressed with dietary salt loading (P < 0.05), with no effect of drug (P > 0.35). 4. In conclusion, blood pressure and renal function were not adversely affected by celecoxib, even during dietary salt loading. These findings support current guidelines suggesting minimal cardiovascular risks associated with short-term, low-dose use of celecoxib in young to middle-aged adults. PMID- 21631569 TI - Epidermal growth factor receptor signalling mediates growth hormone-induced growth of chondrocytes from sex hormone-inhibited adolescent rats. AB - 1. Growth hormone (GH) has been demonstrated to overcome the inappropriate deceleration of growth rate in children with central precocious puberty treated with gonadotropin-releasing hormone analogue (GnRHa). However, the underlying mechanisms remain largely unclear. In the present study, we investigated the potential involvement of the epidermal growth factor receptor (EGFR) pathway in the growth promotion by GH using in vitro cultured growth plate chondrocytes isolated from adolescent rats treated with GnRHa. 2. Chondrocytes were stimulated with GH in the presence or absence of the Janus tyrosine kinase (JAK) 2 inhibitor AG490 (1, 10 and 100 nmol/L), the EGFR kinase inhibitor AG1478 (0.1, 1 and 10 nmol/L), U0126 (an inhibitor of extracellular signal-regulated kinase (Erk) activation; 10 MUmol/L) or a neutralizing antibody against epidermal growth factor (EGF Ab; 0.1, 1 and 10 MUg/mL). The proliferation of chondrocytes was assessed by the 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide assay and immunostaining for proliferating cell nuclear antigen (PCNA). Phosphorylation of Erk1/2 and EGFR was detected by western-blotting. Intracellular mRNA and extracellular protein levels of EGF were detected using reverse transcription-polymerase chain reaction and ELISA, respectively. 3. Growth hormone promoted the proliferation of chondrocytes, which was correlated with increased phosphorylation of Erk1/2 and EGFR and enhanced expression of EGF. Pretreatment with AG490, AG1478, U0126 or EGF Ab completely or partially inhibited the proliferation of chondrocytes and activation of Erk1/2 and EGFR. Pretreatment with AG490, AG1478, or U0126 partially inhibited the expression of EGF. 4. The findings indicate that GH promotes chondrocyte proliferation by activating EGFR signalling. PMID- 21631570 TI - NAMPT -3186C/T polymorphism affects repaglinide response in Chinese patients with Type 2 diabetes mellitus. AB - 1. In the present study, we investigated the associations of nicotinamide phosphoribosyltransferase (NAMPT)-3186 C/T and -948G/T polymorphisms with the risk of Type 2 diabetes mellitus (T2DM) and their impact on the efficacy of repaglinide in Chinese Han T2DM patients. 2. In all, 170 patients with T2DM and 129 healthy controls were genotyped for NAMPT-948G>T and -3186C>T polymorphisms. Thirty-five patients with different NAMPT -3186 C/T genotypes and the same organic anion-transporting polypeptide 1B1 (OATP1B1521) T/C genotype were randomly selected to undergo 8 weeks preprandial repaglinide treatment (1 mg, three times daily). Serum fasting plasma glucose (FPG), post-prandial plasma glucose (PPG), glycated haemoglobin (HbAlc), fasting serum insulin (FINS), post prandial serum insulin (PINS), triglyceride (TG), total cholesterol (CHO), homeostasis model assessment of insulin resistance (HOMA-IR), low-density lipoprotein-cholesterol (LDL-C) and high-density lipoprotein-cholesterol (HDL-C) were determined before and after repaglinide treatment. 3. After repaglinide treatment for 8 consecutive weeks, there were significantly decreases in PFG, PPG, HbAlc, CHO and LDL-C, and increases in FINS, HDL-C and the HDL-C : LDL-C ratio, in T2DM patients. The elevated PINS value in patients with CT genotypes was significantly lower than that in patients with the CC and TT genotypes (P < 0.05) and there were significant differences in CHO between patients with the CT genotype and the CC or TT genotype (P < 0.05). 4. The data suggest that the NAMPT -3186C>T polymorphism is significantly associated with plasma levels of PINS and CHO in Chinese T2DM patients with repaglinide monotherapy. PMID- 21631571 TI - Expression of interleukin-17 is correlated with interferon-alpha expression in cutaneous lesions of lupus erythematosus. AB - BACKGROUND: Type I interferon (IFN) has been reported to have an important role in the development of cutaneous lupus erythematosus (CLE) and systemic lupus erythematosus (SLE). A new subset of CD4+ T cells, T helper (Th)17 cells, also plays a role in the development of autoimmunity. AIM: To investigate expression of interleukin (IL)-17 and IFN-alpha in different CLE subsets, and their associations with the pathogenesis of LE. METHODS: Skin tissue samples from 33 cases, including chronic discoid LE (n = 24), acute (A)CLE (n = 4), subacute CLE (n = 1) and lupus panniculitis (n = 4) were collected for immunohistochemistry. Expression of IL-6, IL-17A, IFN-alpha, IFN-gamma, myxovirus protein (Mx)A and transforming growth factor (TGF)-beta was assessed in these samples. RESULTS: All LE specimens had staining for IL-6 and TGF-beta in the infiltrated inflammatory cells. IL-17A staining was seen in 84.8% of specimens, and IFN-alpha or MxA was seen in 93.9%. TGF-beta expression in ACLE was significantly greater than that in both chronic cutaneous (CC)LE and in lupus panniculitis (P = 0.02 for both). Expression of IL-17A was positively associated with expression of IFN-alpha and MxA (Spearman's rho = 0.56 and 0.39, respectively). In addition, the Cutaneous Lupus Erythematosus Disease Area and Severity Index (CLASI) correlated positively with expression of IFN-alpha and MxA (rho = 0.40 for both), whereas there was no correlation with IL-17A expression. CONCLUSIONS: Two major cytokines, IL-17A and IFN-alpha, may play roles in the pathogenesis of CLE. Their patterns of expression positively correlated with each other. PMID- 21631572 TI - Risk of suicidal ideation in adolescents with both self-asphyxial risk-taking behavior and non-suicidal self-injury. AB - This study examined adolescent participation in self-asphyxial risk-taking behaviors (SAB), sometimes known as the "choking game," and its relationship with other adolescent risk behaviors, including non-suicidal self-injury (NSSI). Researchers proposed that participation in SAB and NSSI would be associated with suicidal behavior, disordered eating, and substance use. Using a large community based sample, results revealed preliminary associations between SAB and other risk-taking behaviors. Adolescents who had engaged in both SAB and NSSI reported more concurrent risk behaviors than adolescents who participated in only one of the behaviors or neither behavior. Results indicate that greater awareness of SAB is important, and continued research can evaluate the possible link between the behavior and risk for suicide. PMID- 21631573 TI - Ecstasy use and suicidal behavior among adolescents: findings from a national survey. AB - The relationship between ecstasy use and suicidal behavior among adolescents in the United States was examined. Data from the adolescent subsample (ages 12-17, N = 19,301) of the 2000 National Household Survey on Drug Abuse were used in the analyses. Information on adolescent substance use, suicidal behaviors, and related sociodemographic, family, and individual factors was obtained in the survey. The rate of past year suicide attempt among adolescents with lifetime ecstasy use was almost double that of adolescents who had used other drugs only, and nine times that of adolescents with no history of illicit drug use. In multinomial logistic regression analyses controlling for related factors, the effect of ecstasy use remained significant. Adolescent ecstasy users may require enhanced suicide prevention and intervention efforts. PMID- 21631574 TI - Completed suicides in Colorado: differences between Hispanics and non-Hispanic Whites. AB - All suicides by Hispanics (n = 434) and non-Hispanic Whites (n = 3,370) in Colorado from 2004 to 2008 using the Violent Death Reporting System were examined. Hispanic victims were significantly younger. Adjusting for age and gender, Hispanic victims were less likely to have reported depressed mood [odds ratio (OR) 0.78; 95% confidence interval (CI) 0.63-0.97], mental health diagnosis (OR 0.53; 95% CI 0.41-0.7), or current psychiatric treatment (OR 0.47; 95% CI 0.43-0.77). There were no differences in reports of financial, relationship, job, or legal stresses. Hispanic suicides were equally likely to be by overdose, firearm, or hanging, but more likely to be in jail (OR 2.68; 95% CI 1.55-4.65). To prevent suicides, stronger partnerships are needed among public health, medical, mental health, and criminal justice professionals. PMID- 21631575 TI - Purpureocillium, a new genus for the medically important Paecilomyces lilacinus. AB - Paecilomyces lilacinus was described more than a century ago and is a commonly occurring fungus in soil. However, in the last decade this fungus has been increasingly found as the causal agent of infections in man and other vertebrates. Most cases of disease are described from patients with compromised immune systems or intraocular lens implants. In this study, we compared clinical isolates with strains isolated from soil, insects and nematodes using 18S rRNA gene, internal transcribed spacer (ITS) and partial translation elongation factor 1-alpha (TEF) sequences. Our data show that P. lilacinus is not related to Paecilomyces, represented by the well-known thermophilic and often pathogenic Paecilomyces variotii. The new genus name Purpureocillium is proposed for P. lilacinus and the new combination Purpureocillium lilacinum is made here. Furthermore, the examined Purpureocillium lilacinum isolated grouped in two clades based on ITS and partial TEF sequences. The ITS and TEF sequences of the Purpureocillium lilacinum isolates used for biocontrol of nematode pests are identical to those causing infections in (immunocompromised) humans. The use of high concentrations of Purpureocillium lilacinum spores for biocontrol poses a health risk in immunocompromised humans and more research is needed to determine the pathogenicity factors of Purpureocillium lilacinum. PMID- 21631576 TI - Molecular characterization of the microbial community in hydrogenetic ferromanganese crusts of the Takuyo-Daigo Seamount, northwest Pacific. AB - The abundance and phylogenetic diversity of the microbial community in the hydrogenetic ferromanganese crust, sandy sediment and overlying seawater were investigated using a culture-independent molecular analysis based on the 16S rRNA gene. These samples were carefully collected from the Takuyo-Daigo Seamount, located in the northwest Pacific Ocean, by a remotely operated vehicle. Based on quantitative PCR analysis, Archaea occupy a significant portion of the prokaryotic communities in the ferromanganese crust and the sediment samples, while Bacteria dominated in the seawater samples. Phylotypes belonging to Gammaproteobacteria and to Marine group I (MGI) Crenarchaeota were abundant in clone libraries constructed from the ferromanganese crust and sediment samples, while those belonging to Alphaproteobacteria were abundant in that from the seawater sample. Comparative analysis indicates that over 80% of the total phylotype richness estimates for the crust community were unique as compared with the sediment and seawater communities. Phylotypes related to Nitrosospira belonging to the Betaproteobacteria and those related to Nitrosopumilus belonging to MGI Crenarchaeota were detected in the ferromanganese crust, suggesting that these ammonia-oxidizing chemolithoautotrophs play a role as primary producers in the microbial ecosystem of hydrogenetic ferromanganese crusts that was formed as precipitates from seawater. PMID- 21631577 TI - Harvesting of novel polyhydroxyalkanaote (PHA) synthase encoding genes from a soil metagenome library using phenotypic screening. AB - We previously reported the construction of metagenomic libraries in the IncP cosmid vector pRK7813, enabling heterologous expression of these broad-host-range libraries in multiple bacterial hosts. Expressing these libraries in Sinorhizobium meliloti, we have successfully complemented associated phenotypes of polyhydroxyalkanoate synthesis mutants. DNA sequence analysis of three clones indicates that the complementing genes are homologous to, but substantially different from, known polyhydroxyalkanaote synthase-encoding genes. Thus we have demonstrated the ability to isolate diverse genes for polyhydroxyalkanaote synthesis by functional complementation of defined mutants. Such genes might be of use in the engineering of more efficient systems for the industrial production of bioplastics. The use of functional complementation will also provide a vehicle to probe the genetics of polyhydroxyalkanaote metabolism and its relation to carbon availability in complex microbial assemblages. PMID- 21631578 TI - Clinical grading systems: can we resolve the needs of clinical practice and those of clinical research? PMID- 21631579 TI - Can lameness be graded reliably? PMID- 21631580 TI - Clinical studies utilising ordinal data: pitfalls in the analysis and interpretation of clinical grading systems. PMID- 21631581 TI - Lower airway disease, now and in the future. AB - Respiratory disease is an important cause of wastage in the Thoroughbred industry and is a cause of welfare problems in all types of horse. This review concerns developments in our understanding of equine respiratory disease since 1998 and illustrates how strategic funding from the Horserace Betting Levy Board has contributed to this understanding. PMID- 21631582 TI - Maternal dexamethasone treatment in late gestation induces precocious fetal maturation and delivery in healthy Thoroughbred mares. AB - REASONS FOR PERFORMING STUDY: The foal requires an active hypothalamo-pituitary adrenal (HPA) axis for organ maturation and post natal survival. Prenatal administration of synthetic glucocorticoids may provide an effective method for inducing fetal maturation safely in the mare. OBJECTIVES: To determine whether dexamethasone administered to late pregnant mares: 1) will induce fetal maturation and precocious delivery; 2) is safe to use and 3) to identify endocrine responses in the mare and foal. METHODS: Pregnant Thoroughbred mares received either 100 mg dexamethasone i.m. (treated n = 5) or 50 ml saline i.m. (control n = 5) at 315, 316 and 317 days of gestation. Plasma progestagens, cortisol and prostaglandin F(2alpha) metabolite (PGFM) concentrations were measured before and after treatment. The foals were weighed, the crown-rump length (CRL) measured and an adrenal stimulation test performed on Day 1. RESULTS: Dexamethasone significantly (P<0.01) reduced gestation length in treated mares without apparent adverse effects. Plasma progestagens increased (P<0.05), and cortisol and PGFM (P<0.05) decreased, following dexamethasone treatment compared with control mares. Foals were clinically mature but those from dexamethasone treated mares had reduced (P<0.05) CRL, but not bodyweights, compared with controls. Their cortisol concentrations increased following exogenous adrenocorticotrophic hormone stimulation but 2 foals from dexamethasone treated mares showed evidence of adrenal suppression. CONCLUSIONS: Dexamethasone stimulates precocious fetal maturation and delivery in healthy late pregnant mares. However, fetal HPA activity may be suppressed. POTENTIAL RELEVANCE: Dexamethasone treatment could be used to improve foal viability in mares at risk of preterm delivery. The endocrine effects of such a therapy must be evaluated before clinical intervention with glucocorticoids can be recommended. PMID- 21631583 TI - Radiological anatomy of the donkey's foot: objective characterisation of the normal and laminitic donkey foot. AB - REASONS FOR PERFORMING STUDY: Anatomical change within a laminitic foot is of diagnostic and prognostic importance. A lateromedial radiograph represents the current 'gold standard' by which these changes are identified. Detection of anatomical change is dependent upon a priori knowledge of normality and subjective assessment alone may not identify modest change. Normal baseline data is, therefore, needed against which objective comparisons can be made. There is little information regarding the radiological anatomy of the donkey foot, hence an equine model has been widely adopted. However, descriptive accounts suggest fundamental anatomical differences between these 2 species. OBJECTIVES: To characterise objectively the radiological anatomy of normal donkey feet and define the nature and extent of anatomical change associated with laminitis. METHODS: The anatomy of the forefoot was quantified from lateromedial radiographs of 83 normal and 74 laminitic donkeys, using a computer based imaging system. Data were analysed using univariate and bivariate statistical methods. RESULTS: Baseline data were established that define the radiological characteristics of the anatomy of normal donkey feet. The key hoof, bone and weightbearing stance parameters of lateromedial radiographs have been evaluated. Laminitis was associated with significant rotation and distal displacement of the distal phalanx, increases in integument depth and morphometric change to the distal phalanx (P<0.05). CONCLUSIONS: This study challenges the validity of applying an equine model to the radiological anatomy of donkey feet. Hence, the diagnosis of anatomical change cannot be based on baseline data previously given for the horse and guidelines should be revised accordingly for the donkey. POTENTIAL RELEVANCE: This study provides an objective basis for the identification of anatomical change associated with laminitis in donkey feet. PMID- 21631584 TI - Selective blockade of A(2A) receptor protects against neurotoxicity induced by kainic acid in young rats. AB - The aim of this study was to investigate the effect of SCH 58261, a selective adenosine A(2A) receptor (A(2A)R) antagonist, on kainic acid (KA)-induced seizures in 21-day-old rats. Rats were pretreated with SCH 58261 (1 or 3 mg/kg) by intraperitoneal (i.p.) route 30 min before KA (10 mg/kg, i.p.) administration. The appearance of clonic seizures, the latency for the onset of the first clonic seizure episode, and the number of deaths induced by KA were evaluated. To test the hypothesis of the oxidative imbalance induced by KA exposure, reactive species (RS) levels, catalase (CAT), glutathione peroxidase (GPx), and glutathione S-transferase (GST) activities in the brains of rats were measured. Both doses of SCH 58261 prolonged the latency for the onset of the first clonic seizure episode. SCH 58261, at the highest dose, decreased the appearance of clonic seizures as well as the mortality rate induced by KA administration. SCH 58261, at the dose of 3 mg/kg, was also effective in protecting against alterations in oxidative stress parameters (RS levels, CAT, GPx, and GST activities) in the brains of young rats exposed to KA. Our data reveal that SCH 58261 was protective against the neurotoxicity induced by KA. Therefore, the blockade of A(2A)R might represent a novel approach for the treatment of seizures. PMID- 21631585 TI - Physicochemical and biological aspects of macrophage-mediated drug targeting in anti-microbial therapy. AB - Macrophages are important drug targets as they mediate a wide variety of infectious diseases. Visceral leishmaniasis (VL), schistosomiasis, brucellosis, and salmonellosis are some of the well-known infectious diseases in which macrophages play a prominent pathophysiological role. For instance, VL parasites exclusively house in the macrophages of liver and spleen. They are resistant to lysosomal degradation by unknown mechanisms, they survive and thrive safely within macrophages, they multiply, and they ultimately affect visceral organs, leading to severe pathological and sometimes even fatal conditions. The majority of routinely used drugs administered in free form distribute all over the body via systemic circulation, leading to relatively low therapeutic activity and a certain degree of toxicity. Unlike for nonmicrobial diseases, targeting parasites procuring resistance and ineffective therapeutic outcome can be obviously speculated in case of infectious disease. The preferential uptake by macrophages, intended to improve the balance between efficacy and toxicity, can be achieved by the use of nanomedicines, i.e. submicron-sized macromolecular or particulate drug delivery systems. This insight has stimulated researchers to use nanomedicines- which tend to be recognized by macrophages as 'foreign' and consequently are taken up by the intended target cells much more effectively than their free drug counterparts--to improve the treatment of infectious diseases. The literature reports extensively on such approaches; however, there are several constraints that limit the application of nanomedicine in macrophage-mediated drug targeting. Here, we briefly describe the strategies that are used to achieve effective drug targeting to macrophages, using VL as a model disease, and we also put forth an understanding of the most important limiting factors. Various physicochemical and biological factors used by researchers as reported in the literature are addressed, and the most important mechanisms and modes by which macrophage specific drug targeting can be achieved are summarized. Based on the evidence obtained to date, it can be concluded that targeting macrophages is a valuable and validated strategy for improving the treatment of infectious diseases. PMID- 21631586 TI - Protective effects of liquiritin apioside on cigarette smoke-induced lung epithelial cell injury. AB - Cigarette smoking is associated with an increased incidence of chronic obstructive pulmonary disease (COPD). In this study, we hypothesized that liquiritin apioside (LA), a main flavonoid component from Glycyrrhiza uralensis, had antioxidant properties by inducing glutathione (GSH) biosynthesis via the inhibition of cytokines and protected lung epithelial cells against cigarette smoke-mediated oxidative stress. A549 cells were treated with cigarette smoke extract (CSE) and/or LA. ICR mice were exposed to cigarette smoke (CS) for four days with increasing exposure time for up to 6 h per day to elicit epithelial cells injury. One hour before smoke exposure, mice were treated with LA by gavage; 18 h after the last CS exposure all examinations were performed. Treatment with LA concentration-dependently prevented CSE-induced cytotoxicity, increase of TGF-beta and TNF-alpha mRNA expression, depletion of GSH and apoptosis in A549 cells. LA at doses 3, 10 and 30 mg/kg dose-dependently inhibited pulmonary neutrophil and macrophage inflammation. Lung sections of the CS-exposed LA treated mice showed an apparently reduced pulmonary inflammation and a significant inhibitory effect on mucus containing goblet cells in the large airways. Furthermore, the CS-induced pulmonary release of TGF-beta, TNF-alpha and myeloperoxidase activity was reduced, and superoxide dismutase activity was enhanced.These results indicate that protective roles of LA on CS-induced the lung epithelial cell injury are mediated by inhibiting TGF-beta and TNF-alpha expression and increasing anti-oxidative levels of GSH, suggesting that LA might be effective as protective agent against epithelial injury in COPD. PMID- 21631587 TI - Sirolimus and everolimus intestinal absorption and interaction with calcineurin inhibitors: a differential effect between cyclosporine and tacrolimus. AB - The mTOR inhibitors (ImTORs) sirolimus (SRL) and everolimus (EVR) have been increasingly used in renal transplantation as part of calcineurin inhibitor (CNI) sparing or avoidance regimens. Those drugs have low and variable oral bioavailability that is increased when combined with cyclosporine or tacrolimus (TAC). We investigated the mechanisms involved in ImTORs intestinal absorption in vitro and associated it with their drug-drug interactions with CNIs. The transport of ImTORs across Caco-2 cells was studied in the apical (A) to basolateral (B) and B to A directions, in the absence or presence of cyclosporine, TAC, and GF120918 (P-gp inhibitor). In Caco-2 cells, EVR and SRL displayed a polarized transport with 8.7- and 5.9-fold higher P(app,B->A) than P(app,A->B), respectively. P-gp inhibition by GF120918 resulted in a 70 and 41% decrease in EVR and SRL efflux, respectively. Cyclosporine and TAC led to a comparable and significant decrease in the efflux ratio of ImTORs, suggesting inhibition of a P-gp-mediated efflux transport. Cyclosporine also exhibited a specific increase of P(app,B->A), which may be attributed to the inhibition of other transporters and/or metabolizing enzymes. In conclusion, EVR and SRL are both subject to an apically directed efflux mediated by P-gp. TAC mainly inhibits this efflux mechanism, while the effect of cyclosporine appears to be more complex with mechanisms to be confirmed by further studies. PMID- 21631588 TI - Hallucinations treated with rivastigmine in Creutzfeldt-Jakob disease. AB - Clinical features at onset of Creutzfeldt-Jakob disease (CJD) may mimic symptoms of Lewy bodies dementia. Clinical evolution, neuroimaging, metabolism exploration, and cerebrospinal fluid investigations may help establishing the diagnosis. However, CJD definite diagnosis requires postmortem autopsy. This symptom overlap led us to successfully prescribe an anticholinesterasic treatment, rivastigmine, to a patient for whom a probable CJD disease was finally diagnosed. PMID- 21631590 TI - Short progression-free survival predicts for poor overall survival in older patients with multiple myeloma treated upfront with novel agent-based therapy. AB - OBJECTIVES: To assess the importance of the quality of response and of early relapse in unselected elderly patients with myeloma treated upfront with novel agents. METHODS: We analyzed 135 unselected transplant-ineligible patients older than 65 yr who were treated upfront with novel agent-based regimens in a single center. RESULTS: On intent to treat, 81% of patients achieved a response (28% sCR/CR, 23% VGPR, and 30% PR). Median progression-free survival (PFS) for patients who achieved sCR/CR was 31 vs. 20 months for VGPR and 23 months for PR (P = 0.048). Median overall survival (OS) for patients with sCR/CR was 62 months, 53 months for VGPR and 38 months for patients with PR (P = 0.028). Early relapse (PFS < 12 months) was more common in patients with PR (39% vs. 21% for VGPR vs. 3% for sCR/CR). Patients who relapsed or progressed < 12 months from initiation of treatment had a median OS of 15.4 months compared with 53 months (P < 0.001) for patients who had a PFS > 12 months despite the fact that after relapse or progression most patients were treated again with novel agents. In multivariate analysis, short PFS was the most significant adverse prognostic factor affecting OS, associated with a 7.25-fold (P < 0.0001) increase in the risk of death. CONCLUSION: In newly diagnosed patients over 65 yr, treated upfront with novel agents achievement of CR and a PFS >= 12 months is associated with improved outcome. Patients who fail to respond or experience early relapse after primary therapy with novel agent-based regimens should be encouraged to participate in clinical trials of novel agents and combinations. PMID- 21631589 TI - Chymotrypsin C mutations in chronic pancreatitis. AB - Chronic pancreatitis is a persistent inflammatory disorder characterized by destruction of the pancreatic parenchyma, maldigestion, and chronic pain. Mutations in the chymotrypsin C (CTRC) gene encoding the digestive enzyme CTRC have been shown to increase the risk of chronic pancreatitis in European and Asian populations. Here, we review the biochemical properties and physiological functions of human CTRC, summarize the functional defects associated with CTRC mutations, and discuss mechanistic models that might explain the increased disease risk in carriers. PMID- 21631591 TI - Measurement of buccal bone volume of dental implants by means of cone-beam computed tomography. AB - PURPOSE: The aim of this study was to evaluate the accuracy of cone-beam computed tomography (CBCT) for measuring the buccal bone volume around dental implants. MATERIALS AND METHODS: Three to six implants were inserted into the anterior maxilla of eight skulls, depending on the availability of bone, and after this, the CBCT was performed. By means of CBCT image, measurements of the bone wall at three points of the implant were obtained, analyzed and compared with those obtained in the plaster skull casting. RESULTS: The results showed that for the three points of the implants, no statistically significant difference in the measurements was obtained from the plaster model and CBCT images. CONCLUSIONS: CBCT can be a useful tool for assessing buccal bone volume along the implant. PMID- 21631592 TI - Masseter muscle thickness, chewing efficiency and bite force in edentulous patients with fixed and removable implant-supported prostheses: a cross-sectional multicenter study. AB - OBJECTIVES: Edentulous patients may be restored with conventional dentures (C/C), implant-supported overdentures (IOD) or implant-supported fixed dental prostheses (IFDP). Null-hypotheses: chewing efficiency, maximum voluntary bite force (MBF) and masseter muscle thickness (MMT) are lower in patients with C/IOD compared with the patients with bimaxillary IFDPs. Both groups perform better than C/C and are inferior to fully dentate controls. MATERIAL AND METHODS: Ethical approval was obtained. For this multicenter cross-sectional study, 80 patients were recruited. Four groups of different dental states comprised of either implant supported prostheses (C/IOD and IFDP/IFDP) or served as control-groups (C/C and fully dentate D/D). Chewing efficiency was assessed with a two-colour mixing ability test. MBF was measured bilaterally with a force gauge. Two dimensional ultrasonography was used to measure MMT bilaterally. RESULTS: Chewing efficiency in C/IOD and IFDP/IFDP (difference NS) was better than in C/C, but not as good as in D/D. MBF in C/IOD was lower than in IFDP/IFDP. Chewing efficiency and MBF were significantly lower in IFDP/IFDP, who had experienced chipping or fracture of the prosthetic superstructure. Median MMT of patients with implant-supported prostheses was between those with C/C and fully dentate participants. There was no significant difference in MMT between C/IOD and IFDP/IFDP. CONCLUSION: Supporting complete prostheses with oral implants seems to have positive effects on the thickness of the masseter muscle, maximum bite force as well as chewing efficiency. The type of implant-supported prostheses may have an influence on the magnitude of the effect. PMID- 21631593 TI - Detecting bone loss along dental implants by subtraction of panoramic radiographs. AB - OBJECTIVES: To test an extended subtraction method for measuring changes in bone along dental implants depicted on non-standardized panoramic radiographs. MATERIAL AND METHODS: The measuring method consisted of a subtraction module extended with modules correcting gray values, correcting geometry and defining the region of interest mesial and distal of dental implants. It was applied to an archive of panoramic radiographs of implant patients who had been monitored up to 16 years. RESULTS: Significant loss of gray value was demonstrated at a constant rate of 0.6 units per month throughout the study. This indicates gradual bone loss on the mesial and/or distal sides of the implants. Females were found to lose bone at a higher rate than males. Smokers and non-smokers, and various implant strategies yielded the same rate of bone loss. CONCLUSIONS: The measuring method is a helpful tool to monitor changes around implants even when non standardized radiographs are being used. Whereas changes of marginal bone level are increased five to eight times during the first year after surgery, the present method shows gradual loss of bone during 15 years after surgery. PMID- 21631594 TI - Calcium phosphate/poly(D,L-lactic-co-glycolic acid) composite bone substitute materials: evaluation of temporal degradation and bone ingrowth in a rat critical sized cranial defect. AB - OBJECTIVES: The present study aimed to provide temporal information on material degradation and bone formation using composite (C) bone defect filler materials consisting of calcium phosphate cement (CaP) and poly(D,L-lactic-co-glycolic acid) (PLGA) microparticles (20 or 30 wt%) in rat critical-sized cranial defects. MATERIALS AND METHODS: Critical-sized bicortical cranial defects were created in 48 rats and CaP/PLGA cement composites were implanted for 4, 8 and 12 weeks (n=8). RESULTS: Histological analysis of the retrieved specimens revealed that implant degradation was significantly faster for C30% (remaining implant up to 89.4 +/- 4.4% at 12 weeks) compared with C20% (remaining implant upto 94.8 +/- 2.1% at 12 weeks), albeit that overall degradation was limited. Although bone formation was limited in both experimental groups (upto 685765.9 MUm(2) for C20% vs. 917603.3 MUm(2) for C30%), C30% showed a significant temporal increase of total bone formation. The percentage of defect bridging was comparable for C20% and C30% at all implantation periods (range 40 +/- 25.5% at week 4 to 65 +/- 20% at week 12 for C20%; range 51.8 +/- 7.8% at week 4 to 70.5 +/- 16.2% at week 12 for C30%). CONCLUSION: The amount of PLGA-microparticles in the CaP/PLGA cement composites demonstrated acceleration of material degradation, while bone formation was found not to be influenced. Further optimization of the composite material is necessary to increase control over degradation and tissue ingrowth. PMID- 21631595 TI - Accuracy of implant casts generated with splinted and non-splinted impression techniques for edentulous patients: an optical scanning study. AB - BACKGROUND: The accuracy of implant casts generated with various impression techniques was mainly investigated in vitro resulting in limited clinical data. PURPOSE: (1) To compare the three-dimensional (3-D) accuracy of splinted and non splinted impression techniques to the control casts (verification jigs) that had been used for actual patient treatment; and (2) to determine the maximum level of clinically undetectable misfit. The null hypothesis was that there would be no significant difference in the accuracy of casts generated with different impression techniques. MATERIALS AND METHODS: The implant casts used for the prosthetic rehabilitation of 12 edentulous jaws with CAD/CAM zirconia, implant fixed complete dental prosthesis (IFCDP) were included in this study. Intraoral acrylic jigs were used to fabricate index casts. Splinted and non-splinted, open tray techniques were used to generate two casts. Optical scanning acquisition of the x-coordinates, y-coordinates and z-coordinates of the implant positions for each individual cast was performed. The "best fit" algorithm was used with computer software to superimpose the scanning datasets. Group I (n=12) included casts from the splinted impression technique vs. acrylic jig casts, and group II (n=12) included casts from non-splinted technique vs. jig casts. RESULTS: The paired t-test and Wilcoxon's signed ranks test were used to compare the 3-D discrepancies within and between groups I (splinted vs. jig) and II (non-splinted vs. jig), respectively. Significant difference was found at the x-axis, y-axis and 3-D between groups I and II (P<0.05), but not in the vertical z-axis (P>0.05). Within subject, global 3-D discrepancies between groups I and II were significantly different (P<0.05), corroborated by in vivo observations of clinical fit. Implant position in the arch affected the 3-D accuracy of casts for both anterior and posterior implants (P<0.05). CONCLUSION: The splinted technique generated more accurate master casts than the non-splinted technique for one piece IFCDPs in edentulous jaws and the null hypothesis was rejected. These clinical implications demonstrate improved accuracy of splinted impression techniques compared with the non-splinted technique. For the external connection, the implant system used in this study, a 3-D misfit ranging from 59 to 72 MUm, may be considered the maximum discrepancy resulting in an acceptable clinical fit with one-piece IFCDPs. PMID- 21631596 TI - Sinus augmentation using absorbable collagen sponge loaded with Escherichia coli expressed recombinant human bone morphogenetic protein 2 in a standardized rabbit sinus model: a radiographic and histologic analysis. AB - OBJECTIVES: The purpose of this pilot study was to determine the osteoinductive effect of absorbable collagen sponge (ACS) loaded with Escherichia coli-expressed recombinant human bone morphogenetic protein 2 (ErhBMP-2) and evaluate structural stability of ACS in a standardized rabbit sinus model. MATERIAL AND METHODS: The maxillary sinuses were prepared bilaterally in six male white rabbits. The windows were prepared using a 6 mm trephine bur, and circular bony windows were carefully removed. Following reflection of the sinus membrane, a saline-soaked ACS alone and an ErhBMP-2-loaded ACS were inserted into the left and right maxillary sinuses, respectively. After a healing period of 8 weeks, sections of the augmented sinus and surrounding bone were made and analyzed by microcomputed tomography and histologically for signs of window closure and bone augmentation. RESULTS: Radiographic analysis revealed new bone formation in both groups of augmented sinus (i.e., with and without ErhBMP-2). The maximum augmented height did not differ significantly between the groups; however, window closure was significantly more advanced in the ErhBMP-2 group than in the control group (P=0.02). The defect was significantly deeper in the control group than in the ErhBMP-2-treated group (P=0.02). CONCLUSIONS: In conclusion, ErhBMP-2-loaded ACS showed enhanced osteoinductive potential, particularly with regard to bone closure of a sinus window and facilitated maturation of the newly formed bone within the rabbit sinus cavity. However, the structural durability of ACS was not sufficient to maintain the augmented volume in the sinus. PMID- 21631597 TI - Implant-supported mandibular overdentures: a cross-sectional study. AB - OBJECTIVE: The aim of this cross-sectional study was to determine the clinical outcome and patient satisfaction in subjects treated with mandibular overdentures supported by two implants. MATERIAL AND METHODS: One hundred and fifty-nine patients, who received restorative therapy in the edentulous mandible consisting of a bar-retained overdenture supported by two osseointegrated implants in a private clinic in Italy, were recalled for a clinical and radiographic examination. One hundred and forty-one subjects with 280 implants attended the examination. The average follow-up time was 3.9 years. The radiographic examination included assessments of the distance between the implant margin and the most coronal position of bone-to-implant contact at the mesial and distal aspects of each implant. A questionnaire regarding comfort, satisfaction with the treatment, aesthetics, speaking capacity and efficiency in chewing was obtained from each subject. Biological and technical complications were recorded and the number of visits due to complications between the delivery of the prostheses and the re-examination was determined. RESULTS: The results from the examination revealed that the number of lost implants was small and the average marginal bone level around the implants was 0.67 mm apical of the implant margin. The most frequently observed complication was hyperplasia of the mucosal tissue under the bar construction. Few patients experienced loosening of retention. The vast majority of patients reported to be satisfied in relation to the restorative therapy from both functional and aesthetic points of view. CONCLUSION: Patients with edentulous mandibles may be successfully rehabilitated by means of two implants supporting a bar-retained overdenture. PMID- 21631598 TI - The use of onplants and implants in children with severe oligodontia: a retrospective evaluation. AB - OBJECTIVE: To evaluate a stable treatment concept for the oral rehabilitation of children with severe oligodontia. The treatment is intended to solve a specific problem caused by limited skeletal growth, multiple congenital missing teeth, underdeveloped bone volume and young age. MATERIAL AND METHODS: A surgical/prosthodontic approach was chosen for six patients with subtotal edentulism despite their young age. To support removable dentures the maxilla was treated with onplants (OnPlant), whereas dental rootform implants (NobelReplace, Nobel Biocare) were placed in the mandible. RESULTS: Eight onplants were placed in the hard palate of four patients for rehabilitating the maxilla and eight dental rootform implants were placed interforaminally in three patients to rehabilitate the mandible. While none of the rootform implants was lost, one onplant was lost and replaced with a stable outcome. The mean follow-up time of the onplants was 5 (+/-1) years vs. 3 (+/-2) years for the rootform implants. CONCLUSION: The evaluation showed the treatment concept described to ensure stable oral rehabilitation and recovery of the stomatognathic system even during the juvenile growth period. PMID- 21631599 TI - Effect of block-periodized exercise training on bone and coronary heart disease risk factors in early post-menopausal women: a randomized controlled study. AB - The purpose of this 12 month randomized exercise intervention was to determine the effect of a block-periodized multipurpose exercise program on bone mineral density (BMD) and parameters of the metabolic syndrome (MetS) in early post menopausal women. Eighty-five subjects (52.3 +/- 2.4 years) living in the area of Erlangen (Germany) were randomly assigned into an exercise (EG, n=43) or a wellness-control group (CG: n=42). The EG performed a periodized multipurpose exercise program with 4-6-week blocks of high-intensity bone-specific exercise intermitted by 10-12 weeks of exercise dedicated to increase endurance and reduce cardiac and metabolic risk factors. The CG performed a low-volume/low-intensity "wellness" program to increase well-being. After 12 months, significant exercise effects were observed for the lumbar spine (LS) BMD as assessed by quantitative computed tomography [total BMD (EG: -0.3 +/- 2.1% vs CG: -2.1 +/- 2.2%, P=0.015); trabecular BMD (EG: -0.7 +/- 3.4% vs CG: -4.7 +/- 4.9%, P=0.001) and dual-energy x-ray absorptiometry (DXA) (EG: -0.1 +/- 2.2% vs CG: -2.0 +/- 2.0%, P=0.002)]. However, no significant effects were observed for total hip BMD as assessed by DXA (P=0.152). Although all MetS parameters were favorably affected among the EG, only the effect for waist circumference was significant. In summary, short periods of bone-specific intervention embedded in longer periods of exercises dedicated to improve cardiovascular and metabolic risk factors positively affected BMD at the LS. PMID- 21631600 TI - Capsular serotype of Staphylococcus aureus in the era of community-acquired MRSA. AB - Capsular polysaccharide (CP) plays an important role in the pathogenicity and immunogenicity of Staphylococcus aureus, yet the common serotypes of S. aureus isolated from US pediatric patients have not been reported. We investigated capsular serotype as well as methicillin susceptibility, presence of Panton Valentine leukocidin (PVL), and clonal relatedness of pediatric S. aureus isolates. Clinical isolates were tested for methicillin susceptibility, presence of mecA, lukS-PV and lukF-PV, cap5 and cap8 genes by PCR, and for capsular or surface polysaccharide expression (CP5, CP8, or 336 polysaccharide) by agglutination. Genetic relatedness was determined by pulsed-field gel electrophoresis. All S. aureus isolates encoded cap5 or cap8. Sixty-nine percent of 2004-2005 isolates were methicillin-susceptible (MSSA) and most expressed a detectable capsule. The majority of MRSA isolates (82%) were unencapsulated, exposing an expressed cell wall techoic acid antigen 336. Pulsed-field type USA300 were MRSA, PVL-positive, unencapsulated strains that were associated with deep skin infections and recurrent disease. Over half (58%) of all isolates from invasive pediatric dermatologic infections were USA300. All pediatric isolates contained either capsule type 5 or capsule type 8 genes, and roughly half of the S. aureus clinical disease isolates from our population were diverse MSSA encapsulated strains. The majority of the remaining pediatric clinical disease isolates were unencapsulated serotype 336 strains of the PVL(+) USA300 community associated-MRSA clone. PMID- 21631601 TI - IFN-gamma plays an essential role in the pathogenesis of gastric lymphoid follicles formation caused by Helicobacter suis infection. AB - In this study, we aimed to assess the role of helper T cells in the development of gastric lymphoid follicles induced by Helicobacter suis infection. C57BL/6J mice were orally inoculated with H. suis. Six weeks after infection, gastric lymphoid follicles were observed in the gastric mucosa by hematoxylin and eosin staining, and the number of follicles was increased throughout the infection period. An immunohistological examination showed that the lymphoid follicles were composed of B cells, CD4-positive helper T cells, and dendritic cells (DC). It was also revealed that the mRNA expression level of interferon-gamma (IFN-gamma) in the gastric mucosa was significantly increased at 12 weeks after infection. No gastric lymphoid follicles were detected in IFN-gamma-deficient mice that had been infected with H. suis at 12 weeks after infection, although the development of lymphoid follicles in IL-4-deficient mice infected with H. suis was similar to that seen in the wild-type mice. In conclusion, IFN-gamma, a Th1 cytokine, is deeply involved in the pathogenesis of gastric lymphoid follicles induced by H. suis infection, and it is suggested that CD4-positive T cells and DC aid in the expansion of gastric lymphoid follicles. PMID- 21631602 TI - Loss of co-chaperone TopJ impacts adhesin P1 presentation and terminal organelle maturation in Mycoplasma pneumoniae. AB - Mycoplasma pneumoniae is a wall-less human respiratory tract pathogen that colonizes mucosal epithelium via a polar terminal organelle having a central electron-dense core and adhesin-related proteins clustered at a terminal button. A mutant lacking J-domain co-chaperone TopJ is non-cytadherent and non-motile, despite having a core and normal levels of the major cytadherence-associated proteins. J-domain co-chaperones work with DnaK to catalyse polypeptide binding and subsequent protein folding. Here we compared features of the topJ mutant with other cytadherence mutants to elucidate the contribution of TopJ to cytadherence function. The topJ mutant was similar ultrastructurally to a non-cytadherent mutant lacking terminal organelle proteins B/C, including aberrant core positioning and cell morphology in thin sections, but exhibited a hybrid satellite growth pattern with features of mutants both having and lacking a core. Time-lapse images of mycoplasmas expressing a YFP fusion with terminal organelle protein P41 suggested that terminal organelle formation/positioning was delayed or poorly co-ordinated with cell growth in the absence of TopJ. TopJ required a core for localization, perhaps involving HMW1. P1 trypsin accessibility on other non-cytadherent mutants was significantly enhanced over wild type but unexpectedly was reduced with topJ mutant cells, suggesting impaired processing, translocation and/or folding of this adhesin. PMID- 21631603 TI - Vanadate and triclosan synergistically induce alginate production by Pseudomonas aeruginosa strain PAO1. AB - Alginate overproduction by P. aeruginosa strains, also known as mucoidy, is associated with chronic lung infections in cystic fibrosis (CF). It is not clear how alginate induction occurs in the wild-type (wt) mucA strains. When grown on Pseudomonas isolation agar (PIA), P. aeruginosa strains PAO1 and PA14 are non mucoid, producing minimal amounts of alginate. Here we report the addition of ammonium metavanadate (AMV), a phosphatase inhibitor, to PIA (PIA-AMV) induced mucoidy in both these laboratory strains and early lung colonizing non-mucoid isolates with a wt mucA. This phenotypic switch was reversible depending on the availability of vanadate salts and triclosan, a component of PIA. Alginate induction in PAO1 on PIA-AMV was correlated with increased proteolytic degradation of MucA, and required envelope proteases AlgW or MucP, and a two component phosphate regulator, PhoP. Other changes included the addition of palmitate to lipid A, a phenotype also observed in chronic CF isolates. Proteomic analysis revealed the upregulation of stress chaperones, which was confirmed by increased expression of the chaperone/protease MucD. Altogether, these findings suggest a model of alginate induction and the PIA-AMV medium may be suitable for examining early lung colonization phenotypes in CF before the selection of the mucA mutants. PMID- 21631604 TI - Inside-out Z rings--constriction with and without GTP hydrolysis. AB - The bacterial tubulin homologue FtsZ forms a ring-like structure called the Z ring that drives cytokinesis. We showed previously that FtsZ-YFP-mts, which has a short amphipathic helix (mts) on its C terminus that inserts into the membrane, can assemble contractile Z rings in tubular liposomes without any other protein. Here we study mts-FtsZ-YFP, where the membrane tether is switched to the opposite side of the protofilament. This assembled 'inside-out' Z rings that wrapped around the outside surface of tubular liposomes. The inside-out Z rings were highly dynamic, and generated a constriction force that squeezed the tubular liposomes from outside. This is consistent with models where the constriction force is generated by curved protofilaments bending the membrane. We used this system to test how GTP hydrolysis by FtsZ is involved in Z-ring constriction. Without GTP hydrolysis, Z rings could still assemble and generate an initial constriction. However, the constriction quickly stopped, suggesting that Z rings became rigidly stabilized in the absence of GTP hydrolysis. We propose that remodelling of the Z ring, mediated by GTP hydrolysis and exchange of subunits, is necessary for the continuous constriction. PMID- 21631605 TI - Regulation of transcription by DNA supercoiling in Mycoplasma genitalium: global control in the smallest known self-replicating genome. AB - The mollicute Mycoplasma genitalium causes sexually transmitted disease in humans. It has recently come to widespread public attention through its involvement in pioneering synthetic biology experiments. The 580-kilo-base-pair genome of M. genitalium contains just 470 genes, few of which seem to encode conventional transcription regulators. This raises the important question of how does this simple organism control its gene expression? The finding that the transcription of an osmotically inducible gene encoding a lipoprotein is sensitive to antibiotics that inhibit the activity of DNA gyrase, the enzyme that introduces negative supercoiling into DNA, raises the possibility that changes in DNA supercoiling provide a regulatory mechanism for controlling transcription in M. genitalium. PMID- 21631606 TI - A mutation within the C-terminal domain of Sup35p that affects [PSI+] prion propagation. AB - The epigenetic factor [PSI+] in the yeast Saccharomyces cerevisiae is due to the prion form of Sup35p. The N-terminal domain of Sup35p (N), alone or together with the middle-domain (NM), assembles in vitro into fibrils that induce [PSI+] when introduced into yeast cells. The Sup35p C-terminal domain (C), involved in translation termination, is essential for growth. The involvement of Sup35p C terminal domain into [PSI+] propagation is subject to debate. We previously showed that mutation of threonine 341 within Sup35p C-domain affects translation termination efficiency. Here, we demonstrate that mutating threonine 341 to aspartate or alanine results in synthetic lethality with [PSI+] and weakening of [PSI+] respectively. The corresponding Sup35D and Sup35A proteins assemble into wild-type like fibrils in vitro, but with a slower elongation rate. Moreover, cross-seeding between Sup35p and Sup35A is inefficient both in vivo and in vitro, suggesting that the point mutation alters the structural properties of Sup35p within the fibrils. Thus, Sup35p C-terminal domain modulates [PSI+] prion propagation, possibly through a functional interaction with the N and/or M domains of the protein. Our results clearly demonstrate that Sup35p C-terminal domain plays a critical role in prion propagation and provide new insights into the mechanism of prion conversion. PMID- 21631607 TI - Lipoamide dehydrogenase is essential for both bloodstream and procyclic Trypanosoma brucei. AB - Lipoamide dehydrogenase (LipDH) is a component of four mitochondrial multienzyme complexes. RNA interference or the deletion of both alleles in bloodstream Trypanosoma brucei resulted in an absolute requirement for exogenous thymidine. In the absence of thymidine, lipdh-/- parasites showed a severely altered morphology and cell cycle distribution. Most probably, in bloodstream cells with their only rudimentary mitochondrion, LipDH is required as component of the glycine cleavage complex which generates methylene-tetrahydrofolate for dTMP and thus DNA synthesis. The essential role of LipDH in bloodstream parasites was confirmed by an in vivo model. Lipdh-/- cells were unable to infect mice. Our data further revealed that degradation of branched-chain amino acids takes place but is dispensable. In cultured bloodstream--but not procyclic--African trypanosomes, the total cellular concentration of LipDH increases with increasing cell densities. In procyclic parasites, LipDH mRNA depletion caused an even stronger proliferation defect that was not reversed by thymidine suggesting that in the fully elaborated mitochondrion of these cells the primary effect is not on the glycine cleavage complex. Since the medium used for the cultivation of procyclic cells was not supplemented with glucose, impairment of the 2 ketoglutarate dehydrogenase complex is probably the main effect of LipDH depletion. PMID- 21631608 TI - Understanding inverse oxygenation responses during motor imagery: a functional near-infrared spectroscopy study. AB - Motor imagery (MI) is described as the mental rehearsal of voluntary movements. We used wireless functional near-infrared spectroscopy (fNIRS) recorded over secondary motor areas during performance of MI and motor execution (ME) in 11 healthy subjects, who either executed or imagined two drawing tasks differing in shape and frequency, i.e. simple (circle, 0.2 Hz) and complex (curved shape, 0.333 Hz). At the group level, results showed that fNIRS is capable of discriminating between the task mode, i.e. MI vs. ME, and the task complexity, i.e. simple vs. complex. At the single-subject level, we observed inverse oxygenation responses, i.e. a decrease in Delta[O(2) Hb] and/or increase in Delta[HHb]. These inverse responses only occurred during MI tasks and were highly correlated, in the first place, with task mode, and secondly with task complexity. Inverse Delta[O(2) Hb] responses are likely to reflect individual differences in performance-related signals and may contribute to the commonly observed inter-subject variability in fNIRS measurements. As MI is now widely used as a mental task in neurorehabilitative applications, the resulting oxygenation pattern may be of use for future developments. For this programme to be successful it is crucial to determine the sources of inter-subject variability. Our study presents a first effort in this direction, indicating that MI-related inverse Delta[O(2) Hb] responses are correlated, first, with task mode and, secondly, with task complexity. PMID- 21631609 TI - Early onset wheeze associated with enhanced combined IL-1beta, IL-6, and IL-12/IL 23p40 in LPS-stimulated cord blood mononuclear cells. AB - BACKGROUND: Neonates with a family history of atopy are at higher risk for developing wheezing in early life. OBJECTIVE: From a birth cohort of at risk infants (first-degree family with atopic disease), we evaluated the influence of distinct intrinsic immunologic risk factors on wheezing disorders in the first 2 years of life. METHODS: Cord blood samples were collected from 195 eligible subjects of a birth cohort of 253 subjects. The subjects studied were those who developed wheezing (n=34) or eczema (n=29) in the first 2 years of life, and 65 healthy control infants. At the time of thawing the viability of the cells were median 70% (range 67.5%-72.5%). Cytokines from lipopolysaccharide (LPS) stimulated mononuclear cells were analysed using fluorescent-activated cell sorting-array and their profiles were evaluated using factor analysis. RESULTS: Infants with wheeze were significantly associated with enhanced combined LPS stimulated IL-1beta, IL-6, and IL-12/IL-23p40 compared with healthy controls (P=0.003). This profile was also associated with the increased risk for wheeze at 2 years of age (OR=2.45; 95% CI=1.50-3.93, P=0.001). LPS-stimulated cytokine IL-8 was also significantly higher in the wheeze group compared with healthy controls and eczema (P=0.003). Intracellular staining showed that monocytes are main producers of IL-6 and IL-8 from cord blood mononuclear cells. Most of the subjects were non-atopic with 3/34 (9%) wheeze and 9/29 (31%) eczema subjects sensitized to the common dietary or inhalant allergens. CONCLUSION AND CLINICAL RELEVANCE: In infants at genetic risk of atopy, wheeze but not eczema in the first 2 years of life is associated with intrinsic hyperresponsive innate cytokine responses which might predispose infants to wheeze development. Distinct pre-symptomatic hyperresponsive innate immune responses risk factors were found to be associated with early onset wheeze disorders, but not eczema. PMID- 21631610 TI - Potential allergens of green gram (Vigna radiata L. Millsp) identified as members of cupin superfamily and seed albumin. AB - BACKGROUND: No systematic study on allergenicity of green gram seed proteins have been performed so far, although incidences of IgE-mediated reaction to green gram seedlings have been reported. OBJECTIVE: We sought to investigate the allergenic potential of green gram, followed by identification and characterization of its relevant allergens using proteomic approaches. METHODS: BALB/c mice were sensitized intraperitoneally with green gram proteins, and levels of specific Igs, Th2 cytokines, histamine, anaphylactic symptoms and histopathological responses were studied. Twelve naso-bronchial allergic patients with a history of sensitization to green gram were selected on the basis of positive skin prick test and elevated specific IgE levels. Green gram allergens were identified and characterized by their ability to endure pepsin, by IgE immunoblot of two dimensional (2D) gels in combination with mass spectrometry and by bioinformatics approaches. RESULTS: Increased specific IgE, IgG1, Th2 cytokine and histamine levels, high anaphylactic scores and histological changes in lungs and spleen of green gram crude protein extract-treated mice are indicative of its sensitization ability. Four proteins (molecular weights: 52, 50, 30 and 18 kDa) showed pepsin resistance and IgE-binding capability with sensitized human and mice sera. The four proteins tentatively named as Vig r2 (52 kDa, pI 5.7), Vig r3 (50 kDa, pI 5.8), Vig r4 (30 kDa, pI 6.6) and Vig r5 (18 kDa, pI 5.5) showed significant sequence similarity with known allergens of soybean, lentil, pea, lupin, etc. Mass spectrometric analysis identified Vig r2 as 8S globulin beta-isoform precursor, Vig r3 as 8S globulin alpha-isoform precursor and Vig r4 as seed albumin. CONCLUSION AND CLINICAL RELEVANCE: Green gram seeds contain at least four clinically relevant allergenic proteins, namely Vig r2, Vig r3, Vig r4 and Vig r5 that were capable of inducing strong IgE-mediated reactions. One of the most important steps towards diagnostic and therapeutic approaches to deal effectively with food allergy is continued identification of newer food allergens and their characterization. The significance of this study can be enormous as the data generated may work as basic biology data in developing a green gram species modified genetically that may have reduced allergenicity. PMID- 21631611 TI - Allergen inhalation challenge in smoking compared with non-smoking asthmatic subjects. AB - BACKGROUND: Smoking asthmatics experience more severe symptoms, require more rescue medication and have more asthma-related hospitalizations than non-smoking asthmatics. However, studies in mice suggest that mainstream cigarette smoke may reduce airway inflammation and may attenuate airway hyperresponsiveness. A comparison of allergen-induced airway inflammatory responses of smoking and non smoking atopic asthmatics has not been examined previously. OBJECTIVES: To determine whether allergen-induced airway responses and inflammatory profiles are attenuated in smoking when compared with non-smoking mild allergic asthmatic subjects. METHODS: Allergen inhalation challenges were performed in 13 smoking and 19 non-smoking mild allergic asthmatic subjects. The forced expired volume in 1 s (FEV(1) ) was measured up to 7 h after allergen inhalation. Methacholine airway responsiveness was measured before and at 24 h after allergen and sputum was induced before and at 7 and 24 h after allergen. RESULTS: Both the smoking and non-smoking groups developed similar allergen-induced falls in FEV(1) during the early and late asthmatic responses and similar increases in allergen-induced airway eosinophils. The mean maximum fall in FEV(1) during the late response was 16.3 +/- 4.3% in non-smokers and 12.9 +/- 7.2% in smokers. The smoking asthmatics, however, did not develop allergen-induced methacholine airway hyperresponsiveness, whereas the non-smoking controls developed a 1.18 doubling dose shift in methacholine PC(20) (P < 0.05). CONCLUSIONS AND CLINICAL RELEVANCE: Mild allergic asthmatic subjects, who were current smokers with a mean 6-year pack history, develop allergen-induced eosinophilic airway inflammation and late responses, similar in magnitude to non-smoking asthmatics, but do not develop methacholine airway hyperresponsiveness associated with the allergen-induced airway eosinophilia. PMID- 21631612 TI - Activin-A: a novel critical regulator of allergic asthma. AB - Activin-A is a pleiotropic cytokine that belongs to the TGF-beta superfamily and plays an important role in fundamental biological processes, such as development and tissue repair. Growing evidence proposes a crucial role for activin-A in immune-mediated responses and associated diseases, with both enhancing and suppressive effects depending on the cell type, the cytokine micromilieu and the context of the response. Several recent studies have demonstrated a striking increase in activin-A expression in experimental models of asthma, as well as, in the asthmatic airway in humans. Importantly, a strong immunoregulatory role for activin-A in allergic airway disease, with suppression of T helper (Th) type 2 cell-driven allergic responses and protection against the development of cardinal features of the asthmatic phenotype was revealed by in vivo functional studies. Activin-A-mediated immunosuppression is associated with induction of functional allergen-specific regulatory T cells. In human asthma, although activin-A levels are increased in the airway epithelium and submucosal cells, the expression of its signalling components is markedly decreased, pointing to decreased regulation. Nevertheless, a rapid activation of the activin-A signalling pathway is observed in the airway of individuals with asthma following inhalational allergen challenge, suggestive of an inherent protective mechanism to control disease. In support, in vitro studies using human airway epithelial cells have demonstrated that endogenous activin-A suppresses the release of inflammatory mediators, while it induces epithelial repair. Collectively, compelling evidence suggests that activin-A orchestrates the regulation of key events involved in the pathogenesis of allergic asthma. The critical role of activin-A in allergic airway responses places this cytokine as an exciting new therapeutic target for asthma. PMID- 21631613 TI - Leukotriene C4 synthase: the engine of aspirin intolerance? PMID- 21631614 TI - Remineralization effects of casein phosphopeptide-amorphous calcium phosphate creme on artificial early enamel lesions of primary teeth. AB - BACKGROUND: Caries in children younger than 72 months is called early childhood caries (ECC). Sixty-six per cent of Chinese children younger than 5 years old have dental decay, and about 97% of them are untreated. AIMS: This in vitro study was conducted to evaluate the remineralization effects of the casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) creme on the artificial early enamel lesions of the primary teeth and to assess its caries-prevention efficiency. DESIGN: Enamel specimens with artificial early lesions were produced and were then randomly divided into Group A: distilled and deionized water, DDW, as negative control; Group B: CPP-ACP creme, test group; Group C: 500 ppm NaF solution, as positive control. The enamel surface microhardness (SMH) was measured before, after demineralization, and 30 days after remineralization. The results were analysed with the SPSS 13.0 software package. The enamel specimens were analysed by the scanning electron microscope. RESULTS: The CPP-ACP creme increased SMH of the eroded enamel significantly more than 500 ppm NaF solution did. The morphology of the enamel was different in each group. CONCLUSIONS: The CPP-ACP creme is effective in remineralizing early enamel lesions of the primary teeth, a little more effective than 500 ppm NaF and can be used for the prevention of ECC. PMID- 21631615 TI - Group integrative reminiscence therapy on self-esteem, life satisfaction and depressive symptoms in institutionalised older veterans. AB - AIMS AND OBJECTIVES: The purpose of this study was to use group integrative reminiscence as a nursing intervention to evaluate the immediate effects on self esteem, life satisfaction and depressive symptoms for a special group named 'institutionalised older veterans' after a 12-week intervention. BACKGROUND: The study group comprised institutionalised older veterans with combat experience, including being wounded in war and who were twice forced to relocate. The group participants had lower life satisfaction, and greater use for mental health services and greater non-specific health complaints were reported from this group. Reminiscence therapy has been considered an effective nursing intervention, but the effects on institutionalised older veterans have not been studied. DESIGN: A quasi-experimental design and purposive sampling were conducted. METHODS: A total of 74 participants were studied with pre- and post tests to measure the effect of group integrative reminiscence therapy. The activity was held once weekly for 12 weeks. The Life Satisfaction Index A, self esteem scale and Geriatric Depression Scale Short Form were used as research tools, and the t-test, Fisher's exact test and generalised estimating equation were used for data analysis. RESULTS: All participants were male, with an average age of 81.34 years old, 91.9% unmarried and were in bad health. After 12 weeks of intervention, the reminiscence groups significantly improved their self-esteem and life satisfaction and decreased depressive symptoms compared with control groups. CONCLUSION: Group integrative reminiscence revealed immediate effects on improving the self-esteem and life satisfaction of institutionalised older veterans, and depressive symptoms were also decreased. Moreover, a sense of positive self-value and belonging to the institution was produced. RELEVANCE TO CLINICAL PRACTICE: Group integrative reminiscence is an applicable nursing intervention for vulnerable persons such as institutionalised older veterans. A structured protocol based on the characteristics of the residents and the aim of the intervention can lead nurses to provide appropriate reminiscent activities. PMID- 21631616 TI - Boundary matters: clinical leadership and the distinctive disciplinary contribution of nursing to multidisciplinary care. AB - AIMS: To describe Irish nurses' views of clinical leadership and to describe their clinical leadership development needs. BACKGROUND: Nurses are often unclear about the precise nature of clinical leadership and its impact on the processes and outcomes of care and little is known about their self-perceived clinical leadership development needs. DESIGN: Seventeen focus group interviews were conducted with a purposive sample of 144 nurses from 13 practice settings. A conceptual lens was provided by the work of Bernstein and Young who emphasise the epistemological, practical and relational significance of boundaries and how they relate in fundamental ways to professionals' sense of their distinctive disciplinary identities and membership of specialised communities of practice. METHODS: Focus group data were collected using semi-structured topic guides. Analysis was facilitated by NVivo 7(c) and interpretation was informed by a conceptual framework arising from the interplay of emerging themes and the literature review. RESULTS: The implications for clinical leadership development of two critical concepts, 'representing nursing' and 'compensatory action', are discussed in detail. CONCLUSIONS: Clinical leadership development should emphasise the development of all nurses as clinical leaders in the context of the delineation, clarification and articulation of their distinctive contribution in multidisciplinary care settings. RELEVANCE TO CLINICAL PRACTICE: Clinical leaders are recognised as practice experts and as leaders in their particular fields. Recognition and influence in and beyond the immediate context of care depends greatly on their ability to articulate the distinct nursing contribution to patient care. This ability provides an essential resource to resist the ongoing blurring, effacement and dilution of nurses' roles. PMID- 21631617 TI - Social gradient in initiation and transition to daily use of tobacco and cannabis during adolescence: a retrospective cohort study. AB - AIMS: This study explores whether the family socio-economic status (F-SES) and school situation could have an impact on tobacco and cannabis initiation and transition to daily use during adolescence. DESIGN AND SETTING: A French cross sectional nation-wide survey conducted in 2005 containing retrospective data. PARTICIPANTS: French teenagers aged 17 (n = 29 393). MEASUREMENT: The F-SES was defined by the highest occupational category of either parent, with seven categories ranging from unemployed/inactive to managers/professionals. Ages at repeat school years, at leaving school, at the first episode of drunkenness and at initiation of illicit drug use were used to model tobacco and cannabis initiation and transition to daily use with time-discrete logistic regressions. FINDINGS: The risk for tobacco initiation was almost equally distributed across F SES groups, but the risk of a progression to daily use was higher in every F-SES category compared to managers/professionals [odds ratio (OR) from 1.17 to 1.90]. Compared to managers/professionals, risk of cannabis initiation was lower in all F-SES categories (OR from 0.63 to 0.87), but all categories except farmers were at increased risk of transition to daily use: the OR range between 1.29 (intermediate) and 1.98 (unemployed/inactive). Repeating school years and leaving school predicted daily use of tobacco (OR = 2.00 and 2.37) and cannabis (4.58 and 2.07). CONCLUSIONS: Adolescents from the highest family socio-economic status categories are at risk for tobacco and cannabis experimentation but are less prone to engage in daily use. Psychological and social mechanisms that inhibit transition to daily use should be investigated, including school attainment and performance. PMID- 21631618 TI - Hypothalamic-pituitary-adrenal axis reactivity to social stress and adolescent cannabis use: the TRAILS study. AB - AIMS: To investigate the relationship of life-time and repeated cannabis use with hypothalamic-pituitary-adrenal (HPA) axis reactivity to social stress in a general population sample of adolescents. DESIGN: Adolescents who reported life time or repeated cannabis use, life-time or repeated tobacco use and never use of either cannabis or tobacco were compared with respect to their HPA axis reactivity during the Groningen Social Stress Task (GSST), which was based on the Trier Social Stress Task. SETTING: A large prospective population study of Dutch adolescents [the TRacking Adolescents' Individual Lives Survey (TRAILS) study]. PARTICIPANTS: A total of 591 adolescents (51% male) who participated in the GSST, which was an additional measurement during the third assessment wave. MEASUREMENTS: HPA axis stress-reactivity was indexed by four cortisol samples collected before, during and after the GSST. Furthermore, all adolescents in our study completed self-reported questionnaires on life-time and repeated cannabis and tobacco use. Models were adjusted for sex, recent alcohol use, experimental session risk status, socio-economic status, mood and time of the experimental session. FINDINGS: Life-time cannabis users had significantly lower stress reactivity levels when compared to abstainers [odds ratio (OR) = 0.68, confidence interval (CI) = 0.55-0.85, P < 0.01] and life-time tobacco users (OR = 0.79, CI = 0.64-0.98, P < 0.05). In addition, repeated cannabis users also exhibited lower stress-reactivity levels when compared to life-time ever users of either tobacco or cannabis (OR = 0.74, CI = 0.53-0.98, P < 0.05). CONCLUSIONS: Lower hypothalamic-pituitary-adrenal-axis stress-reactivity in adolescents is related specifically to life-time and repeated cannabis use. PMID- 21631619 TI - Global research neglect of population-based approaches to smoking cessation: time for a more rigorous science of population health interventions. AB - It has been argued that the preponderance of studies into individual smoking cessation therapies seems grossly out of proportion to the number of people who use these therapies to quit smoking, and that this imbalance is due to factors such as the role of the pharmaceutical industry in funding research and a general bias towards individual- rather than population-based approaches to medical and health problems. We believe that there are other significant factors that affect the balance of research in smoking cessation, such as the higher standards of evidence required to justify the implementation of individual medical therapies compared with population-based interventions. We argue that research practitioners in the area of population tobacco control are well placed to address this imbalance by setting more rigorous standards of evidence for population health interventions. This could be achieved by setting aside a small proportion of funds from population health and advocacy activities to invest in studying their effectiveness. We believe that this would potentially return information of sufficient value to justify increasing overall population investment beyond the cost of the additional research component. Additional benefits would be gained from increased research in this area, such as better understanding of how to translate tobacco control initiatives to developing countries with high smoking rates, and how to target disadvantaged and marginalized populations more effectively in developed countries that continue to have high rates of smoking and low rates of smoking cessation, despite the existence of broad population-based strategies. PMID- 21631620 TI - A policy-oriented review of strategies for improving the outcomes of services for substance use disorder patients. AB - AIMS: To inform policy makers on available options for improving the effectiveness of treatments for substance use disorders and to stimulate debate about treatment improvement strategies among public officials, clinical providers, care managers, service users, families and researchers. METHODS: We draw on the scientific literature and our public policy experiences in two countries (the United Kingdom and the United States) to give an overview of policies which may improve care for individuals with substance use disorders. We divide such policies into 'process-focused quality improvement strategies' that attempt to change some aspect of treatment (e.g. increased retention, greater use of evidence-based practices) and 'patient-focused strategies' that attempt to reward outcomes directly (e.g. contingency management for patients, payment by results for providers). FINDINGS: Many policies of both types are poorly developed, have shown poor results, or both. The evidence is clear that process focused quality improvement strategies can change what providers do and how treatment programs work, but such changes have thus far demonstrated only minimal impact on patient outcomes. Patient-focused strategies face challenges including treatment providers avoiding hard-to-treat patients or spending inordinate time relocating patients after treatment to assess outcome. However, policies that reward in-treatment outcomes and policies that allow the patient to purchase desired recovery support services show more promise. As policy makers go forward in this endeavor, they can do an enormous service to their countries and the field by embedding careful evaluation studies alongside new treatment outcome improvement initiatives. PMID- 21631621 TI - DSM-IV to DSM-5: the impact of proposed revisions on diagnosis of alcohol use disorders. AB - AIMS: To determine the prevalence of past 12-month DSM-5 alcohol use disorders (AUDs), to quantify and characterize individuals who remain stably unaffected or affected and those who 'switch' diagnostically between DSM-IV and DSM-5 classifications. DESIGN: Data from the nationally representative wave 2 of the National Epidemiological Survey of Alcohol and Related Conditions (NESARC) collected in 2004-05. SETTING: General population survey. PARTICIPANTS: All surveyed participants (n=34,653, aged 21 years and older) and 29,993 individuals reporting life-time alcohol use across both waves of NESARC. MEASUREMENTS: DSM-IV and DSM-5 criteria were coded using proposed guidelines. FINDINGS: The prevalence of DSM-5 AUDs was 10.8% with the corresponding prevalence of DSM-IV abuse/dependence being 9.7%, implying a modest 11.3% increase. Those who switched diagnostically from affected to unaffected (19.6% of DSM-IV affected) were most likely to have endorsed hazardous use, due particularly to drinking and driving, while those who transitioned from unaffected to affected (3.3% of DSM-IV unaffected) were primarily DSM-IV diagnostic orphans reporting larger/longer and quit/cut-back. Dropping the legal criterion did not affect the prevalence significantly, while the addition of craving also had a relatively modest impact on prevalence. CONCLUSION: The proposed DSM-5 revisions eliminate successfully individuals diagnosed previously with DSM-IV alcohol abuse due primarily to hazardous use alone and incorporate diagnostic orphans into the diagnostic realm. Definitions of craving and importantly, hazardous use require considerable attention as it is likely that they will contribute to variations in reports of increased prevalence of alcohol use disorders between DSM-IV to DSM-5. PMID- 21631622 TI - Treatment outcomes of a stage 1 cognitive-behavioral trial to reduce alcohol use among human immunodeficiency virus-infected out-patients in western Kenya. AB - AIMS: Dual epidemics of human immunodeficiency virus (HIV) and alcohol use disorders, and a dearth of professional resources for behavioral treatment in sub Saharan Africa, suggest the need for development of culturally relevant and feasible interventions. The purpose of this study was to test the preliminary efficacy of a culturally adapted six-session gender-stratified group cognitive behavioral therapy (CBT) intervention delivered by paraprofessionals to reduce alcohol use among HIV-infected out-patients in Eldoret, Kenya. DESIGN: Randomized clinical trial comparing CBT against a usual care assessment-only control. SETTING: A large HIV out-patient clinic in Eldoret, Kenya, part of the Academic Model for Providing Access to Healthcare collaboration. PARTICIPANTS: Seventy five HIV-infected out-patients who were antiretroviral (ARV)-initiated or ARV eligible and who reported hazardous or binge drinking. MEASUREMENTS: Percentage of drinking days (PDD) and mean drinks per drinking days (DDD) measured continuously using the Time line Follow back method. FINDINGS: There were 299 ineligible and 102 eligible out-patients with 12 refusals. Effect sizes of the change in alcohol use since baseline between the two conditions at the 30-day follow-up were large [d=0.95, P=0.0002, mean difference=24.93, 95% confidence interval (CI): 12.43, 37.43 PDD; d=0.76, P=0.002, mean difference=2.88, 95% CI: 1.05, 4.70 DDD]. Randomized participants attended 93% of the six CBT sessions offered. Reported alcohol abstinence at the 90-day follow-up was 69% (CBT) and 38% (usual care). Paraprofessional counselors achieved independent ratings of adherence and competence equivalent to college-educated therapists in the United States. Treatment effect sizes were comparable to alcohol intervention studies conducted in the United States. CONCLUSIONS: Cognitive-behavioral therapy can be adapted successfully to group paraprofessional delivery in Kenya and may be effective in reducing alcohol use among HIV-infected Kenyan out-patients. PMID- 21631623 TI - Statistical analysis of daily smoking status in smoking cessation clinical trials. AB - AIMS: Smoking cessation trials generally record information on daily smoking behavior, but base analyses on measures of smoking status at the end of treatment (EOT). We present an alternative approach that analyzes the entire sequence of daily smoking status observations. METHODS: We analyzed daily abstinence data from a smoking cessation trial, using two longitudinal logistic regression methods: a mixed-effects (ME) model and a generalized estimating equations (GEE) model. We compared results to a standard analysis that takes abstinence status at EOT as outcome. We evaluated time-varying covariates (smoking history and time varying drug effect) in the longitudinal analysis and compared ME and GEE approaches. RESULTS: We observed some differences in the estimated treatment effect odds ratios across models, with narrower confidence intervals under the longitudinal models. GEE yields similar results to ME when only baseline factors appear in the model, but gives biased results when one includes time-varying covariates. The longitudinal models indicate that the quit probability declines and the drug effect varies over time. Both the previous day's smoking status and recent smoking history predict quit probability, independently of the drug effect. CONCLUSION: When analysing outcomes of studies from smoking cessation interventions, longitudinal models with multiple outcome data points, rather than just end of treatment, can makes efficient use of the data and incorporate time varying covariates. The generalized estimating equations approach should be avoided when using time-varying predictors. PMID- 21631624 TI - Non-medical prescription use increases the risk for the onset and recurrence of psychopathology: results from the National Epidemiological Survey on Alcohol and Related Conditions. AB - AIMS: Given the rising rates of the non-medical use of prescription medications (NUPM) and strong cross-sectional associations between psychopathology and NUPM, we examined whether a history of NUPM increased the risk for onset and recurrence of psychopathology. DESIGN: Longitudinal data are from waves 1 and 2 of the National Epidemiological Survey on Alcoholism and Related Conditions (NESARC). SETTING: The NESARC is a nation-wide, household-based survey. PARTICIPANTS: A total of 34,653 US resident participants who completed both NESARC waves were included in analyses. MEASUREMENTS: Primary measures were history of NUPM and Axis I psychopathology at wave 1. Wave 1 personality disorder diagnosis and sociodemographic characteristics were used as control variables. FINDINGS: Design based logistic regression analyses indicated that life-time and past year NUPM at wave 1 increased risk for onset of psychopathology among those with no history of psychopathology at baseline, with particular risk for non-NUPM substance use and bipolar disorders. Life-time and past year NUPM were associated with recurrence of alcohol and non-NUPM substance use disorders among those with these diagnoses at baseline. In contrast, life-time and past year NUPM was associated with the onset of all examined disorders among those with a different diagnosis at baseline. CONCLUSIONS: Non-medical use of prescription medications is a consistent risk factor for the onset and recurrence of psychopathology. PMID- 21631625 TI - Reducing the legal blood alcohol concentration limit for driving in developing countries: a time for change? Results and implications derived from a time-series analysis (2001-10) conducted in Brazil. AB - AIMS: In Brazil, a new law introduced in 2008 has lowered the blood alcohol concentration limit for drivers from 0.06 to 0.02, but the effectiveness in reducing traffic accidents remains uncertain. This study evaluated the effects of this enactment on road traffic injuries and fatalities. DESIGN: Time-series analysis using autoregressive integrated moving average (ARIMA) modelling. SETTING: State and capital of Sao Paulo, Brazil. PARTICIPANTS: A total of 1,471,087 non-fatal and 51,561 fatal road traffic accident cases in both regions. MEASUREMENTS: Monthly rates of traffic injuries and fatalities per 100,000 inhabitants from January 2001 to June 2010. FINDINGS: The new traffic law was responsible for significant reductions in traffic injury and fatality rates in both localities (P<0.05). A stronger effect was observed for traffic fatality ( 7.2 and -16.0% in the average monthly rate in the State and capital, respectively) compared to traffic injury rates (-1.8 and -2.3% in the State and capital, respectively). CONCLUSIONS: Lowering the blood alcohol concentration limit in Brazil had a greater impact on traffic fatalities than injuries, with a higher effect in the capital, where presumably the police enforcement was enhanced. PMID- 21631626 TI - Alcoholics Anonymous attendance, decreases in impulsivity and drinking and psychosocial outcomes over 16 years: moderated-mediation from a developmental perspective. AB - AIMS: To examine whether decreases in impulsivity account for links between Alcoholics Anonymous (AA) attendance and better drinking and psychosocial outcomes, and whether these mediational 'effects' are conditional on age. DESIGN: A naturalistic study in which individuals were assessed at baseline, and 1, 8 and 16 years later. SETTING: Participants initiated help-seeking through the alcohol intervention system (detoxification programs, information and referral centers). PARTICIPANTS: Individuals with alcohol use disorders and no prior history of substance abuse treatment at baseline [n=628; 47% women; mean age=34.7 years (standard deviation=9.4)]. MEASUREMENTS: Self-reports of impulsivity and drinking pattern at baseline and year 1, duration of AA (number of weeks) in year 1 and drinking (alcohol use problems, self-efficacy to resist drinking) and psychosocial outcomes (emotional discharge coping, social support) at baseline and follow-ups. FINDINGS: Controlling for changes in drinking pattern, decreases in impulsivity were associated with fewer alcohol use problems, better coping and greater social support and self-efficacy at year 1, and better coping and greater social support at year 8. Decreases in impulsivity statistically mediated associations between longer AA duration and improvements on all year 1 outcomes and indirect effects were moderated by participant age (significant only for individuals 25 years of age or younger). CONCLUSIONS: Decreased impulsivity appears to mediate reductions in alcohol-related problems over 8 years in people attending Alcoholics Anonymous. PMID- 21631627 TI - Detection of 2-hydroxyglutarate in formalin-fixed paraffin-embedded glioma specimens by gas chromatography/mass spectrometry. AB - Mutations in the isocitrate dehydrogenase (IDH) 1 and 2 genes occur frequently in diffuse astrocytoma and oligodendroglioma. The consecutive amino acid substitutions in the mutant proteins result in a gain of the function to catalyze the reduction of alpha-ketoglutarate to 2-hydroxyglutarate (2HG). So far, all investigated IDH mutations share this gain of function. We here describe a method to detect 2HG levels in archival formalin-fixed paraffin-embedded tumor specimens by stable isotope dilution using gas chromatography followed by mass spectrometry (GC/MS). While 2HG levels are notably decreased during the routine embedding process, preserved amounts are still sufficient to indicate a mutation. Detection of 2HG in archival specimens could make routinely processed tissue accessible for research on 2HG accumulation and may allow studies on correlation with clinical data. PMID- 21631628 TI - DNA hypermethylation and histone modifications downregulate the candidate tumor suppressor gene RRP22 on 22q12 in human gliomas. AB - RRP22 (Ras-related protein on chromosome 22) has been suggested as a candidate tumor suppressor in human cancers. Investigating a panel of 70 human gliomas, we found a frequent decrease in the RRP22 mRNA expression levels (67%), preferentially in high-grade gliomas [World Health Organization (WHO) grades III and IV] as compared with low-grade gliomas (WHO grade II). Moreover, reduced RRP22 mRNA expression was associated with shorter overall survival in 180 glioblastoma patients included in the National Institutes of Health Repository for Molecular Brain Neoplasia Data (NIH REMBRANDT) database. Decreased RRP22 expression levels were in part explained by 5'-CpG island hypermethylation and increased by the treatment with the demethylating agent 5-aza-2'-deoxycytidine in glioblastoma cell lines. In addition, the in vitro treatment with the histone deacetylase inhibitor trichostatin A alone resulted in RRP22 reexpression as well as a significant increase in the levels of RRP22 promoter DNA bound to pan acetylated histone H3 and H4. Moreover, in primary human glioblastomas, we observed an increase of H3K9me3-bound and a decrease of pan-Ac-H3-bound RRP22 in comparison with non-neoplastic brain tissue, consistent with a heterochromatinization of the RRP22 promoter. Taken together, our findings demonstrate that both 5'-CpG island hypermethylation and histone modifications contribute to the frequent and prognostically unfavorable transcriptional downregulation of RRP22 in malignant gliomas. PMID- 21631629 TI - Fracture resistance of ceramic veneers with different preparation designs. AB - PURPOSE: The purpose of this study was to examine the fracture load of ceramic veneers with different preparation designs. MATERIALS AND METHODS: Seventy-five extracted, intact, human maxillary central incisors were prepared according to five preparation designs (P) (n: 15) as follows: (1) P2e: 2-mm incisal reduction, preparation entirely in enamel; (2) P4e: 4-mm incisal reduction, preparation entirely in enamel; (3) P2d: 2-mm incisal reduction, preparation entirely in dentin; (4) P4d: 4-mm incisal reduction, preparation entirely in dentin; and (6) Pc: Unrestored, intact teeth as control. All preparations had a butt joint incisal finish line, rounded internal line angles, and cervical finish lines 1 mm above the cementoenamel junction. Ceramic veneers were fabricated with IPS Empress (Ivoclar Vivadent AG, Schaan, Liechtenstein) and cemented with Syntac Classic Adhesive system and Variolink II (Ivoclar) resin cement. Veneers were loaded until fracture at a 90 degrees angle to the lingual surface of the test tooth following the thermocycling process (5 degrees to 55 degrees , 3500 times). Statistical analyses were performed using analysis of variance (ANOVA) and Tukey's Multiple Range Test. RESULTS: The mean fracture loads (SD) were (in N) as follows: (1) P2e: 262 (63); (2) P4e: 189 (40); (3) P2d: 239 (53); (4) P4d: 162 (36); and (5) Pc: 277 (66). The amount of incisal reduction exhibited a significant influence on fracture resistance regardless of the preparation depth (p < 0.05). CONCLUSIONS: Ceramic veneers with preparation designs entirely on dentin with 4-mm incisal reduction yielded lower fracture loads than those prepared with 2-mm incisal reduction. Veneers with 2-mm incisal reduction exhibited fracture resistance similar to that of intact teeth for preparation designs supplied on both enamel and dentin. PMID- 21631630 TI - Dimensional changes in gypsum fragments bonded with cyanoacrylate. AB - PURPOSE: Accidental fractures may occur during manipulation and transportation of plaster casts. In clinical practice, plaster fragments may be bonded without harming the accuracy of the final denture, provided that the bonding agent does not cause dimensional alterations. Cyanoacrylate could be a good material because of its ease of use, quick set, wide availability, and low cost. The aim of this study was to assess the dimensional alteration of Type IV plaster fragments bonded with a cyanoacrylate-based adhesive. MATERIALS AND METHODS: Ten hexagonal regular prisms were made of Type IV plaster, with two reference marks on one of the faces. The distance between the marks was measured under a comparison microscope. After this, the prisms were fractured so that the fracture line would be between the two reference marks, bonded with a cyanoacrylate-based universal adhesive and measured again. RESULTS: The mean difference between the measurements performed before and after fracture and bonding of the fragments was 0.0194 mm. At a level of significance of 0.05, there was no statistically significant difference between the measurements before and after fracture and bonding of the dies (p = 0.1582). CONCLUSION: It may be concluded that bonding of Type IV plaster fragments with a cyanoacrylate-based adhesive did not cause significant dimensional alterations. PMID- 21631631 TI - Treatment options for the replacement of missing mandibular incisors. AB - The replacement of a mandibular incisor is a dental treatment warranting special consideration. Some of the challenges associated with the anterior mandible are limited space, challenging surrounding anatomy, and tough esthetic requirements. Proper diagnosis and treatment planning may require a multidisciplinary approach to successfully meet the demands of replacing a missing tooth in this sextant. Several treatment options currently exist for mandibular incisor replacement. These options include (1) resin-bonded fixed dental prostheses (RBFDPs), (2) orthodontic treatment, (3) full-veneer fixed dental prostheses (FDPs), (4) dental implants for single-tooth replacement, (5) possible extraction of one or more incisors and restoration with implant-supported FDPs, (6) possible extraction of one or more teeth and restoration with FDPs from #22 to 27, (7) possible extraction of one or more teeth and restoration with removable dental prostheses (RDPs). This manuscript outlines the various treatment options for the replacement of mandibular incisors and discusses benefits and drawbacks of each. PMID- 21631632 TI - Breast metastasis from rhabdomyosarcoma of the nasal septum in a pregnant adult woman. PMID- 21631634 TI - Photodynamic therapy with methyl-aminolevulinate for the treatment of double extramammary Paget's disease. PMID- 21631635 TI - Surgical treatments for lentigo maligna: a review. AB - BACKGROUND: Since its initial description by Jonathan Hutchinson 120 years ago, a substantial amount of research has occurred to determine the optimum surgical therapy for lentigo maligna (LM). OBJECTIVE: To summarize the literature regarding the surgical treatment of LM. METHODS: We searched the National Library of Medicine using Pubmed Central and MEDLINE and included as many investigational reports regarding LM therapy that were available in an attempt to form a comprehensive review of surgical modalities. The key words "lentigo maligna,""lentigo maligna treatment,""lentigo maligna therapy," and "lentigo maligna therapeutic modalities" were used. RESULTS: We included 12 studies examining staged surgical excision (SSE), nine using Mohs micrographic surgery (MMS), six investigating cryosurgery, 22 investigating imiquimod, seven using lasers, nine investigating radiation therapy, and two investigating electrosurgery and curettage. CONCLUSIONS: SSE and MMS are associated with the lowest recurrence rates for LM. Cryotherapy and radiation therapy may be considered the options for treatment of LM in patients who cannot tolerate surgery. Imiquimod, although not currently approved by the FDA, has shown some efficacy in limited experimental studies and may play a future role in the treatment of LM. PMID- 21631636 TI - Merkel cell carcinoma in the setting of chronic lymphocytic leukemia. AB - BACKGROUND: Merkel cell carcinoma (MCC) is a rare, aggressive, primary skin cancer that usually occurs in elderly white people on the sun-exposed areas of skin. Chronic lymphocytic leukemia (CLL) is a low-grade, clonal, B-cell, lymphoproliferative disorder that also usually affects older adults. CLL has been associated with multiple other secondary malignancies, especially skin cancer. OBJECTIVE: To further understand the characteristics and behavior of MCC in the setting of concomitant CLL. MATERIALS AND METHODS: We identified patients with diagnoses of both MCC and CLL who were seen at the Mayo Clinic (Rochester, MN) from January 1, 1976, to December 31, 2008, and retrospectively reviewed the charts of the identified patients. RESULTS: Seven patients received diagnoses of MCC and CLL. Five of these patients appeared to have higher rates of recurrence, metastasis, and mortality than patients with MCC but not CLL. CONCLUSIONS: MCC may behave more aggressively in the setting of concomitant CLL, but more studies are needed to definitively evaluate the characteristics and behavior of MCC in these patients. PMID- 21631637 TI - The efficacy of stereoimage optical topometry to evaluate depressed acne scar treatment using cultured autologous fibroblast injection. AB - BACKGROUND: Despite the use of several evaluation methods, the lack of a true consensus scale hinders standardization of evaluation and treatment of depressed acne scars. OBJECTIVE: To investigate the usefulness of a stereoimage optical topometer (SOT) system in evaluating the efficacy of autologous cultured fibroblast injection for depressed acne scars and to compare the results with those obtained using a visual grading system. MATERIALS AND METHODS: Twenty-two patients with depressed acne scars were included. Three doses of cultured autologous fibroblast (2 * 10(7) cell/mL) injections were given at 2-week intervals. Efficacy was evaluated at 2, 4, 12, and 16 weeks using a visual grading system and a SOT. RESULTS: After autologous fibroblast injection, the lesions evaluated using the visual grading and SOT systems steadily improved. Visual grading and SOT variables obtained showed a similar tendency, although an interrater reliability test of the visual grading system correlated poorly. CONCLUSIONS: A three-dimensional morphologic study can be a useful and reliable method of evaluating the severity of acne scars. We were able to more objectively and quantitatively evaluate acne scar severity through combined analysis using visual grading and SOT systems. PMID- 21631638 TI - Photoinduced collagen cross-linking: a new approach to venous insufficiency. AB - BACKGROUND: What little research has been done on methods of venous valve function recovery with radiofrequency has had disappointing results. Valvuloplasty has some supporters, but the majority of physicians do not consider it a valid therapeutic option. OBJECTIVE: To test a new method of treating varicose veins based on their collagen structure. This procedure it is not a thermal treatment, but it is fast, with significant shrinking and preservation of the endothelium. MATERIALS & METHODS: In the laboratory, we subjected greater saphenous vein specimens to irradiation with a blue light-emitting diode generated (wavelength 450-480 nm) while a riboflavin solution (vitamin B2) was administered. The riboflavin acts as a cross-linking agent, and the blue light as the activator. In this photo-induced reaction, oxygen singlet is produced with oxidative deamination, forming new covalent bonds between collagen fibrils and water. RESULTS: In venous specimens, we demonstrated fast and significant shrinkaged without histologic evidence of endothelial damage and with evident change in mechanical properties of varicose veins. CONCLUSIONS: Photochemically induced collagen cross-linking to restructure varicose veins is only a research field but may become an important tool for recovery of vein diameter and valve function. PMID- 21631639 TI - Excisional surgery (scalpectomy) for dissecting cellulitis of the scalp. PMID- 21631640 TI - Mycobacterial peritonitis: difference between non-tuberculous mycobacteria and Mycobacterium tuberculosis. AB - Unlike tuberculous peritonitis, peritonitis due to non-tuberculous mycobacteria (NTM) has unclear clinical manifestations. This study aimed to clarify the clinical manifestations and laboratory results of NTM peritonitis and compare it to tuberculous peritonitis. This retrospective study was conducted from 2000 to 2008 in a medical centre in Taiwan. Patients with mycobacteria isolated from ascites were identified and compared according to causative pathogens (Mycobacterium tuberculosis or NTM). Those with NTM peritonitis were further classified into the 'probable' and 'possible' groups based on diagnostic evidence. Twenty-five patients with NTM peritonitis and 65 with tuberculous peritonitis were reviewed. Mycobacterium avium complex was the most common NTM pathogen (52%). There was no obvious difference between the 'probable' and 'possible' NTM peritonitis groups regarding age and laboratory data. Patients with NTM peritonitis and those with tuberculous peritonitis had no differences in age or gender but varied in symptoms and serum laboratory data. NTM peritonitis was 100% associated with underlying co-morbidities and had lower proportions of lymphocytes and albumin level in ascites. Twelve (48%) NTM peritonitis and 21 (32%) tuberculous peritonitis patients died during the 6-month follow-up. Anti mycobacterial treatment, but not mycobacterial species, was correlated with better 6-month survival. In Taiwan, NTM is responsible for 28% of mycobacterial peritonitis cases, which have a poor prognosis if untreated. There are some differences in clinical manifestations between NTM and tuberculous peritonitis. NTM peritonitis should be considered in patients with peritonitis but without causative microorganisms identified other than NTM. PMID- 21631641 TI - The ins and outs of Mycobacterium tuberculosis drug susceptibility testing. AB - Drug susceptibility testing of Mycobacterium tuberculosis in the diagnostic laboratory classifies clinical isolates as either drug-'resistant' or drug 'susceptible', on the basis of their ability to grow in the presence of a 'critical concentration' of the test compound. From knowledge of the mechanisms that underlie drug resistance, it has become evident that drug resistance in M. tuberculosis is quite heterogeneous and involves low-level, moderate-level and high-level drug resistance phenotypes. Different mutations are associated with different levels of phenotypic resistance, and the acquisition of a genetic alteration leading to a decrease in drug susceptibility does not inevitably exclude the affected compound from treatment regimens. As a result, the simple categorization of clinical M. tuberculosis isolates as 'resistant' on the basis of susceptibility testing at 'critical concentrations' may need to be revised and supplemented by quantitative measures of resistance testing to reflect the biological complexity of drug resistance, with the view of optimally exploiting the compounds available for treatment. PMID- 21631642 TI - Update on the prevention, diagnosis and management of cytomegalovirus infection during pregnancy. AB - Human cytomegalovirus (CMV) is the leading cause of congenital infection, with morbidity and mortality at birth and sequelae. Each year approximately 1-7% (Rev Med Virol 2010; 20: 311) of pregnant women acquire a primary CMV infection. Of these, about 30-40% transmit infection to their fetuses. The risk of serious fetal injury is greatest when maternal infection develops in the first trimester or early in the second trimester. Between 10 and 15% of congenitally infected infants are acutely symptomatic at birth and most of the survivors have serious long-term complications. Until a few years ago, laboratory testing was not possible to precisely define the maternal immune status, the recent development of advanced serological tests (IgG avidity test, IgM immunoblot and neutralizing antibody testing) allow us to identify, among pregnant women with suspected CMV, those with primary infection who are therefore at high risk of transmitting CMV to the fetus. This is done with the use of a screening test. As most maternal infections are asymptomatic, the only way to disclose primary infection is to implement specific serological testing as early in pregnancy as possible (before week 12-16 of gestation). Given the high risk of mother-fetus transmission and fetal damage, prenatal diagnosis is recommended to women with primary CMV infection contracted in the first half of pregnancy and in case of fetal abnormalities suggestive of infection. The correct interpretation of serological and virological tests followed by appropriate counselling by an expert physician is an effective tool to reduce the number of unnecessary pregnancy terminations by over 70%. PMID- 21631643 TI - Metronomic doxifluridine chemotherapy combined with the anti-angiogenic agent TNP 470 inhibits the growth of human uterine carcinosarcoma xenografts. AB - Uterine carcinosarcoma is a highly aggressive gynecological neoplasm that responds poorly to conventional chemotherapy and radiotherapy. Metronomic chemotherapy is accepted as a new approach for cancer treatment, and its underlying mechanism seems to involve the suppression of angiogenesis. However, the efficacy of metronomic and anti-angiogenic therapies against uterine carcinosarcoma is unknown. The anti-angiogenic effect of doxifluridine was assessed in vitro using human umbilical vein endothelial cells (HUVEC) co cultured with FU-MMT-1 human uterine carcinosarcoma cells. The antitumor and anti angiogenic effects of metronomic doxifluridine (delivered via oral gavage) in combination with TNP-470 were evaluated in vivo. Tumor vascularity was assessed by contrast-enhanced color Doppler ultrasound, laser Doppler and microvessel density staining. Doxifluridine suppressed tube formation of HUVEC and vascular endothelial growth factor production by FU-MMT-1 cells. Metronomic doxifluridine alone significantly suppressed tumor growth compared with the untreated (control) group, while metronomic doxifluridine in combination with TNP-470 significantly inhibited tumor growth compared with each treatment alone. A significant reduction of intratumoral vascularity was observed in FU-MMT-1 xenografts following treatment with metronomic doxifluridine in combination with TNP-470, as compared with each treatment alone. Intestinal bleeding was only observed when the maximum tolerated dose of doxifluridine was administered in combination with TNP-470. Metronomic doxifluridine chemotherapy in combination with TNP-470 might be effective for uterine carcinosarcoma without marked toxicity, possibly acting via its potent anti-angiogenic effects. Clinical studies are needed to evaluate the safety and efficacy of this treatment in humans. PMID- 21631644 TI - Gene status of head and neck squamous cell carcinoma cell lines and cetuximab mediated biological activities. AB - Cetuximab is a chimeric IgG1 monoclonal antibody that targets epidermal growth factor receptor (EGFR). Cetuximab binds to EGFR and prevents phosphorylation of EGFR. Moreover, preclinical results have shown the ability of cetuximab to induce either complement-mediated tumor cell killing (CDC) or antibody-dependent cell mediated-cytotoxicity (ADCC). We previously reported mutation in EGFR regarding head and neck squamous cell carcinoma (HNSCC) cell lines. In the present study, we analyzed the same 16 HNSCC cell lines for mutations in KRAS, PIK3CA, BRAF and PTEN. Furthermore, we evaluated cetuximab-mediated biological activities (antiproliferative effect by the MTT assay and ADCC) regarding these cell lines. Mutations in PIK3CA and PTEN were observed in two cell lines (2/16, 12.5%), but no mutation was observed in KRAS and BRAF. The antiproliferative effect of cetuximab by the MTT assay was not strong, and no correlation was observed between the antiproliferative effect of cetuximab and mutations in EGFR, KRAS, PIK3CA, BRAF and PTEN in these cell lines. Therefore, the mutation status of EGFR and downstream molecules were not useful for predicting the antitumor effects of cetuximab on HNSCC. Cetuximab-mediated ADCC was observed in these cell lines and might have been influenced by the expression of EGFR. Therefore, cetuximab mediated ADCC seems to be an important part of the antitumor mechanisms of cetuximab and the expression levels of EGFR might influence the antitumor activity of cetuximab. Therefore, besides the antiproliferative effect of cetuximab by the MTT assay, it appeared important to evaluate cetuximab-mediated ADCC as well as EGFR expression in HNSCC cells. PMID- 21631645 TI - Etiology and treatment of ischaemic stroke in patients with beta-thalassemia major. AB - BACKGROUND AND PURPOSE: Although hypercoagulability-induced thromboembolism is generally accepted as cause of cerebral ischaemia in thalassemic patients, cardiogenic embolism has been recently suggested as another possible stroke etiology. METHODS: We present four adult beta-thalassemia major patients with manifest cardiac involvement who suffered territorial strokes. RESULTS: In the presence of siderotoxic cardiomyopathy and arrhythmia, we assumed cardiogenic embolism as etiology of stroke and initiated oral anticoagulation as preventive medication. Two of our patients were the first beta-thalassemia major patients who underwent successful thrombolysis with rtPA. CONCLUSIONS: Cardioembolism seems to be the cause of stroke in cases of beta-thalassemia major. Thrombolysis can be applied in the setting of acute brain ischaemia in such high risk patients. PMID- 21631646 TI - Medical and non-medical correlates of carpal tunnel syndrome in a Taiwan cohort of one million. AB - BACKGROUND: Carpal tunnel syndrome (CTS), with unclear etiology, is the most common entrapment neuropathy. Its occurrence is related to lots of medical and non-medical conditions with uncertain causality. With a large population, we characterized selected demographical and clinical factors to add more information on CTS-correlated factors and new insight into future CTS prevention. METHODS: A national insurance claim dataset of one million enrollees in Taiwan was used to identify 15 802 patients with CTS and 31 604 randomly selected controls, during a period of 7 years starting 1 January 2000. Statistical association with CTS was determined for five sociodemographic and nine medical factors. RESULTS: Patients were predominantly women (65.6% vs. 47.7% in the control group) and older (40 and above, 62.6% vs. 36.2%). Rheumatoid arthritis was found to be the most significant comorbidity associated with CTS, followed by gout, hypertension, diabetes, obesity, uremia, and acromegaly. For younger group age <=39, the association of these comorbidities was stronger, and hypothyroidism and vitamin B(6) deficiency were additional comorbidities. Aging appears to reduce the relative impact of the diseases commonly associated with CTS as the possible risk factors. CONCLUSIONS: Identification of the CTS correlates in younger group would be of greater value in timely detection and treatment for these diseases. Correcting these disorders may aid in removing possible causes of CTS. This is the first report on the effect of aging on probable CTS risk factors. How factors associated with aging contribute to the development of CTS remains to be determined. PMID- 21631647 TI - Disease severity affects quality of life of hereditary spastic paraplegia patients. AB - BACKGROUND AND PURPOSE: Hereditary spastic paraplegia (HSP) causes progressive gait disturbance because of degeneration of the corticospinal tract. To assess its impact on Health-Related Quality of Life (HRQoL), we analyzed the correlation of HRQoL with disease severity and clinical symptoms in HSP. METHODS: HRQoL was assessed by the Short-Form 36 (SF-36) Mental and Physical Component summary scores (MCS and PCS) in 143 German patients with HSP. Disease severity was assessed by the Spastic Paraplegia Rating Scale (SPRS) and landmarks of walking ability. Patients with 'pure' or 'complicated' HSP were compared. RESULTS: Higher SPRS scores indicating higher disease severity correlated significantly with lower PCS (r = -0.63; P < 0.0005) and MCS (r = -0.38; P < 0.0005) scores. MCS and PCS were reduced in patients with 'complicated' forms compared to 'pure' HSP and with decreasing walking ability. CONCLUSION: HRQoL is substantially impaired in patients with HSP and decreases with disease severity and the presence of 'complicating' symptoms. Patients are most affected by the physical restraints of their disease, but mental health is impaired as well. HRQoL is a valid parameter in HSP that should be considered in upcoming therapeutical trials. PMID- 21631648 TI - Usefulness of MRA-DWI mismatch in neuroendovascular therapy for acute cerebral infarction. AB - BACKGROUND: This study evaluated the usefulness of MR angiography (MRA)-diffusion weighted imaging (DWI) mismatch in neuroendovascular therapy over 3 h after onset of acute cerebral infarction. METHODS: The subjects were 14 cases (age, 73 +/- 8.4 years) who had an anterior circulation deficit on DWI/MRA on arrival and underwent neuroendovascular therapy over 3 h after onset. MRA-DWI mismatch (MDM) (+) was defined as 'major artery lesion (+) and diffusion-weighted image-Alberta Stroke Program Early CT Score (DWI-ASPECTS) >=6'; MDM (-) was defined as 'major artery lesion (+) and DWI-ASPECTS <6'. RESULTS: Reperfusion was achieved in nine of 14 patients (64%) undergoing neuroendovascular therapy. Within the reperfusion group, in the five MDM (+) patients and the four MDM (-) patients, the outcome was a favorable clinical response in the MDM (+) group. The modified Rankin Scale (mRS) scores after 90 days were 0-2 in 3 (60%) and 3-6 in 2 (40%) of the MDM (+) group patients and 0-2 in 0 (0%) and 3-6 in 4 (100%) of the MDM (-) group patients. In the MDM (+) group, a good outcome was achieved. However, the number of cases was small, so this was not a significant difference. Within the non reperfusion group, in the three MDM (+) patients and the two MDM (-) patients, the mRS scores after 90 days were 0-2 in 1 (33%) and 3-6 in 2 (67%) of the MDM (+) group patients and 0-2 in 0 (0%) and 3-6 in 2 (100%) of the MDM (-) group patients. In both groups, the outcome was poor. CONCLUSIONS: With neuroendovascular therapy, a good outcome with reperfusion was achieved in the MDM (+) group compared to the MDM (-) group. This suggests that the presence or absence of MDM may be useful in determining prognosis after reperfusion. PMID- 21631649 TI - Miller Fisher syndrome: brief overview and update with a focus on electrophysiological findings. AB - Miller Fisher syndrome (MFS), a variant of the Guillain-Barre syndrome (GBS), is characterized by ophthalmoplegia, ataxia, and areflexia. The annual incidence is around one patient per one million population. The antiganglioside anti-GQ1b IgG antibody has a role in the pathogenesis of the syndrome, especially of ophthalmoplegia. The presence of this antibody in the serum can be identified in over 80% of the patients, peaking in the first week, whereas albuminocytological dissociation in the cerebrospinal fluid (CSF) appears later. The most consistent electrophysiological findings in MFS are reduced sensory nerve action potentials and absent H reflexes. More variability is seen with F waves and various investigations involving cranial structures. Although there are usually no abnormalities in MFS by routine neuroimaging, in a few cases, contrast enhancement of nerve roots and signs of central nervous system involvement were described supporting the hypothesis of an anti-GQ1b-syndrome, a continuum involving GBS, MFS, and Bickerstaff's brainstem encephalitis. Owing to the lack of randomized trials, treatments used for GBS (intravenous immunoglobulin and plasmapheresis) are usually applied, although from retrospective analyses, the outcome was similar between treated and untreated subjects. The outcome of MFS is usually good with case fatality of < 5%. In the few autopsy cases, macroscopic abnormalities were generally not seen in the nervous system. Microscopic examination of the peripheral nervous system (including cranial nerves) showed segmental demyelination with minimal perivascular infiltration with normal spinal cord and brain stem. PMID- 21631650 TI - Earthquakes and neurological care. PMID- 21631651 TI - The prediction of functional dependency by lateralized and non-lateralized neglect in a large prospective stroke sample. AB - BACKGROUND AND PURPOSE: Lateralized and non-lateralized impairments in visual attention have been identified as important components in patients with visuospatial neglect. This study investigated the course of these two phenomena across time in relation to neurological symptoms and functional outcome in a large consecutive and prospective stroke sample. METHODS: A total of 375 consecutive stroke patients were divided into three groups (lateralized, non lateralized or no visual inattention) acutely and 3 months post-stroke using the star cancellation test. Neurological impairments, localization of brain damage, asymmetry in clinical symptoms and functional outcome were assessed. Possible group differences were analysed, and stepwise logistic regressions were performed to examine the relative importance of predictors of functional dependency. RESULTS: Participants with acute lateralized inattention differed (P <= 0.05) from the other two groups by more often exhibiting severe neurological symptoms, functional dependency and persisting visual inattention. The regression analyses selected acute lateralized inattention as an important and independent predictor of functional dependency following right hemisphere damage, but not following left hemisphere damage. CONCLUSIONS: The results emphasize the prognostic value of lateralized inattention and the importance of separating lateralized and non lateralized symptoms of visual inattention at the commencement of rehabilitation. PMID- 21631652 TI - Neuropsychological outcome after a first symptomatic ischaemic stroke with 'good recovery'. AB - BACKGROUND: Neuropsychological impairment after stroke when no motor, sensory or language deficits are left remains understudied. The primary aim of this study was to assess neuropsychological outcome in a specific population of patients after a first symptomatic stroke without previous cognitive decline and with a good motor, linguistic, and functional recovery (i.e. 'good outcome'). The secondary aims were to identify the profile of this potential impairment and relations between brain lesions and neuropsychological outcome. METHODS: Sixty consecutive patients were evaluated by a comprehensive neuropsychological assessment focusing specifically on executive and attentional functions but also on memory 109 days, on average, after the infarct. Patients were compared with 40 healthy controls matched for age and education. RESULTS: Patients showed lower performance in every cognitive domain compared with controls. Along with an important executive deficit, patients were also impaired on attention and memory. Patients were not more depressed than controls, although they were more apathetic. We also found a significant positive correlation between cognitive impairment and pre-existing white matter brain lesions assessed by MRI. CONCLUSIONS: We report the first study examining the impact of a first stroke on cognition but also on psychiatric disorders in patients with good functional outcome. We found that patients considered as asymptomatic were, in fact, exhibiting a multidomain cognitive deficit that could impact return to life as before stroke. PMID- 21631653 TI - Development of the outcome expectancy scale for self-care among periodontal disease patients. AB - RATIONALE: The theory of self-efficacy states that specific efficacy expectations affect behaviour. Two types of efficacy expectations are described within the theory. Self-efficacy expectations are the beliefs in the capacity to perform a specific behaviour. Outcome expectations are the beliefs that carrying out a specific behaviour will lead to a desired outcome. OBJECTIVE: To develop and examine the reliability and validity of an outcome expectancy scale for self-care (OESS) among periodontal disease patients. METHODS: A 34-item scale was tested on 101 patients at a dental clinic. Accuracy was improved by item analysis, and internal consistency and test-retest stability were investigated. Concurrent validity was tested by examining associations of the OESS score with the self efficacy scale for self-care (SESS) score and plaque index score. Construct validity was examined by comparing OESS scores between periodontal patients at initial visit (group 1) and those continuing maintenance care (group 2). RESULTS: Item analysis identified 13 items for the OESS. Factor analysis extracted three factors: social-, oral- and self-evaluative outcome expectancy. Cronbach's alpha coefficient for the OESS was 0.90. A significant association was observed between test and retest scores, and between the OESS and SESS and plaque index scores. Further, group 2 had a significantly higher mean OESS score than group 1. CONCLUSION: We developed a 13-item OESS with high reliability and validity which may be used to assess outcome expectancy for self-care. A patient's psychological condition with regard to behaviour and affective status can be accurately evaluated using the OESS with SESS. PMID- 21631654 TI - Confidence in receiving medical care when seriously ill: a seven-country comparison of the impact of cost barriers. AB - OBJECTIVE: This paper examines how negative experiences with the health-care system create a lack of confidence in receiving medical care in seven countries: Australia, Canada, Germany, The Netherlands, New Zealand, the United Kingdom, and the United States. METHODS: The empirical analysis is based on data from the Commonwealth Fund International Health Policy Survey 2007, with nationally representative samples of adults aged 18 and over. For the analysis of the experience of cost barriers and confidence in receiving medical care, we conducted pairwise comparisons of group percentages as well as country-wise multivariate logistic regression models. RESULTS: Individuals who have experienced cost barriers show a significantly lower level of confidence in receiving safe and quality medical care than those who have not. This effect is most pronounced in the United States, where people who have foregone necessary treatment because of costs are four times as likely to lack confidence as individuals without the experience of cost barriers (adjusted odds ratio 4.00). In New Zealand, Germany, and Canada, individuals with the experience of cost barriers are twice as likely to report low confidence compared with those without this experience (adjusted odds ratios of 1.95, 2.19 and 2.24, respectively). In The Netherlands and UK, cost barriers are only a marginal phenomenon. CONCLUSIONS: The fact that the experience of financial barriers considerably lowers confidence indicates that financial incentives, such as private co payments, have a negative effect on overall public support and therefore on the legitimacy of health-care systems. PMID- 21631655 TI - The human metapneumovirus: a case series and review of the literature. AB - Human metapneumovirus (hMPV) is an emerging human pulmonary pathogen that is genetically related to respiratory syncytial virus. It has been increasingly associated with respiratory illnesses over the last few decades. Immunocompromised patients are particularly susceptible with resultant morbidity and mortality. We describe our experience with 9 immunocompromised patients diagnosed with pneumonia secondary to hMPV, 2 of whom were successfully treated with aerosolized and oral ribavirin along with intravenous immunoglobulin (IVIG). We suggest that hMPV should be considered in the differential diagnosis of immunocompromised patients with acute respiratory illness. Ribavirin (oral and aerosolized) with IVIG is potentially an effective treatment option for those with severe disease. PMID- 21631656 TI - Small-gauge vitrectomy in traumatic retinal detachment. AB - PURPOSE: To report the outcome of retinal detachment surgery following trauma treated with small-gauge vitrectomy. DESIGN: Retrospective chart review. PARTICIPANTS: Adult patients presenting with a retinal detachment following severe trauma between 2007 and 2009 that underwent vitreoretinal surgery with either 25- or 23-gauge vitrectomy. METHODS: Data collected included; age, sex, eye, time interval between injury and diagnosis of retinal detachment, anatomical and functional success, complications, and length of follow up. MAIN OUTCOME MEASURES: Anatomical and functional outcome. RESULTS: Nineteen consecutive adult patients with traumatic retinal detachment were included. The mean age was 51.4 +/- 19.3 years; 68.5% were male. Thirteen patients (68.4%) presented with visual acuity of hand movement or worse including six patients with light perception only. Five eyes achieved 6/60 vision or better, and three eyes had visual acuity of 6/12 or better. During the period of follow up six eyes experienced a recurrent retinal detachment. CONCLUSION: Traumatic retinal detachment can be successful managed with small-gauge vitrectomy. PMID- 21631657 TI - Periocular steroids in conjunctival reactive lymphoid hyperplasia, a new approach? PMID- 21631658 TI - Deep microsporidial keratitis after keratoconjunctivitis. PMID- 21631659 TI - Microperimetry evidence of functional improvement after vitrectomy for optic disc pit-related intraretinal fluid without serous detachment. PMID- 21631660 TI - Preservation of myelinated nerve fibres in advanced glaucoma. PMID- 21631661 TI - Evaluation and management of mechanical globe injury: comment. PMID- 21631663 TI - Eye health promotion in Western Pacific island countries. PMID- 21631664 TI - Iridotomy in Caucasians: a more conventional angle. PMID- 21631666 TI - Primary orbital squamous cell carcinoma. PMID- 21631667 TI - Extent of asymmetry and unilaterality among juvenile onset primary open angle glaucoma patients. AB - BACKGROUND: Juvenile onset open angle glaucoma (JOAG) due to its rarity is not well characterized. We aimed to assess the extent of interocular asymmetry of baseline intraocular pressure (IOP), disc morphometry and visual field defects at presentation in patients with primary JOAG. DESIGN: Retrospective, single centre, hospital-based study. PARTICIPANTS: Fifty-two consecutive JOAG patients who presented with glaucomatous optic neuropathy in at least one eye, without any secondary causes of glaucoma. METHODS: Participants were evaluated for baseline clinical features. The optic disc parameters were measured using scanning laser ophthalmoscopy (Heidelberg Retina Tomograph). Reliable and reproducible visual field tests using standard 30-2 Humphrey automated perimetry were analysed. MAIN OUTCOME MEASURES: Interocular asymmetry of baseline IOP, vertical cup:disc ratio and mean deviation. RESULTS: Fourteen patients (27%) had glaucomatous optic neuropathy in only one eye at initial presentation; the fellow eyes of which had IOP <21 mmHg in eight whereas six had IOP >21 mmHg. In 20 out of 52 patients (39%) one eye remained perimetrically unaffected. Patients presenting with bilateral optic neuropathy were found to be significantly younger (24.4 +/- 10.6 years) in age compared to those with unilateral optic neuropathy (32 +/- 8.2 years) (P = 0.02). CONCLUSIONS: One-fourth of primary JOAG patients present as a unilateral optic neuropathy with 60% of these having normal IOP in the fellow eyes. Primary JOAG may present with considerable asymmetry with a small proportion presenting as a unilateral disease. PMID- 21631668 TI - Preoperative visual acuity of people undergoing cataract surgery in rural and urban Nepal. AB - BACKGROUND: Preoperative presenting visual acuity (PPVA) is not a commonly reported indicator for the delivery of cataract services. This study aimed to evaluate the PPVA of patients undergoing cataract surgery in rural and urban areas of Nepal. DESIGN: A prospective, non-interventional study conducted from October 2007 to March 2008 in a tertiary hospital and outreach clinics of Nepal. PARTICIPANTS: A total of 3023 consecutive subjects from urban and rural settings (1323 from the tertiary hospital and 1700 from outreach clinics) with cataract were included. METHODS: Visual acuity was tested with a Snellen E-chart at 6 m by ophthalmic assistants. All Outreach Microsurgical Eye Clinic patients were examined by either ophthalmologists or senior ophthalmic assistants. Patients at the Tertiary Eye Care Centre were examined only by ophthalmologists. MAIN OUTCOME MEASURES: Preoperative presenting visual acuity of patients undergoing cataract surgery was measured in both the settings. RESULTS: The sex ratio was 0.96 (male/female). Based on PPVA, 11.5% of patients operated were blind (PPVA < 3/60 in the better eye). The proportion of blind people operated was similar in rural (11.4%) to urban (11.6%) areas. The proportion of blind eyes operated was higher in rural (50.7%) compared to urban (38.9%) areas. CONCLUSION: Despite a comparatively high volume of cataract surgery carried out in Nepal every year, only about one in eight patients operated for cataract in the sample population was blind. PPVA is an important indicator for the progress towards eliminating cataract blindness. PMID- 21631669 TI - Economic impact of primary open-angle glaucoma in Australia. AB - BACKGROUND: Glaucoma is the World's leading cause of irreversible blindness, and poses serious public health and economic concerns. DESIGN: Review. SAMPLES: Published randomized trials and population-based studies since 1985. METHODS: We report the economic impact of primary open-angle glaucoma and model the effect of changes in detection rates and management strategies. MAIN OUTCOME MEASURES: The cost-effectiveness of different interventions to prevent vision loss from primary open-angle glaucoma was measured in terms of financial cost (Australian dollars) and disability-adjusted life years. RESULTS: The prevalence of glaucoma in Australia is expected to increase from 208 000 in 2005 to 379 000 in 2025 because of the aging population. Health system costs over the same time period are estimated to increase from $AU355 million to $AU784 million. Total costs (health system costs, indirect costs and costs of loss of well-being) will increase from $AU1.9 billion to $AU4.3 billion in Australia. CONCLUSION: Primary open-angle glaucoma poses a significant economic burden, which will increase substantially by 2025. This dynamic model provides a valuable tool for ongoing policy formulation and determining the economic impact of interventions to better prevent visual impairment and blindness from glaucoma. PMID- 21631670 TI - Comparison of standard trabeculectomy versus microtrabeculectomy as a surgical treatment for glaucoma: a randomized clinical trial. AB - BACKGROUND: To determine the effect of scleral flap size on the medium-term intraocular pressure control and complication rates after augmented trabeculectomy. DESIGN: Prospective randomized clinical trial. PARTICIPANTS: Glaucoma patients undergoing primary trabeculectomy. Exclusion criteria included previous ocular surgery apart from cataract surgery, secondary glaucoma and age under 18. METHODS: Patients were randomized to either standard trabeculectomy (4 * 4 mm scleral flap) or microtrabeculectomy (2 * 2 mm scleral flap), both with adjustable sutures and antimetabolites. Bleb needling was performed as required. Patients were evaluated at day 1, weeks 1, 3, 6 and months 3, 6, 12, 18 and 24 postoperatively. MAIN OUTCOME MEASURES: Vision, intraocular pressure, complications and failure (intraocular pressure >= 21 mmHg or not reduced by >=20% from baseline, intraocular pressure <= 5 mmHg, repeat glaucoma surgery and no light perception vision). RESULTS: Forty-one patients were recruited; 20 had standard trabeculectomy, and 21 had microtrabeculectomy. At 2 years, the mean intraocular pressure and cumulative probability of failure was 12.4 +/- 4.6 mmHg and 0.28 for standard trabeculectomy, and 11.5 +/- 3.6 mmHg and 0.27 for microtrabeculectomy (P = 0.50 and 0.89, respectively). One patient in each group required Baerveldt device implantation. Vision reduced >=2 Snellen lines in 15% in the standard trabeculectomy group and 25% in the microtrabeculectomy group, mainly from cataract (P = 0.48). CONCLUSION: Both trabeculectomy techniques achieved good intraocular pressure reduction and had similar complication rates. Scleral flap size had no significant effect on medium-term intraocular pressure control and complication profile. PMID- 21631671 TI - Late form of early capsular block syndrome. PMID- 21631672 TI - Mannose-binding lectin as part of the complement pathway: characterization in non inflamed and inflamed human eyes. AB - BACKGROUND: Mannose-binding lectin plays a central effector role in the lectin pathway of complement activation. Frequently occurring MBL2 polymorphisms result in mannose-binding lectin deficiency, which increases susceptibility to infection. We characterized mannose-binding lectin levels and function in non inflamed and inflamed human eyes, and evaluated its relationship to blood mannose binding lectin levels and function. DESIGN: Prospective, observational clinical study with controls and cases. PARTICIPANTS: Twenty-seven patients with paired blood and ocular samples (aqueous and/or vitreous) including 15 controls (non inflamed) and 12 cases (inflamed). METHODS: Blood and ocular samples were collected from controls (n = 15) with quiet eyes during elective cataract surgery and cases with inflamed eyes including proven/suspected endophthalmitis (n = 11) and herpetic retinal vasculitis (n = 1). Mannan-binding and C4 deposition enzyme linked quantify mannose-binding lectin levels and function. MAIN OUTCOME MEASURES: Blood and ocular mannose-binding lectin levels and function. RESULTS: Of 27 patients, 10 (37%) were mannose-binding lectin-deficient (defined as blood mannose-binding lectin levels <500 ng/mL). Blood mannose-binding lectin levels (P= 0.16) or function (P= 0.43) were not significantly different between controls and cases. As expected, there was a high correlation between blood mannose binding lectin levels and function (r(2) = 0.74). However, there was significantly more mannose-binding lectin in inflamed eyes than non-inflamed eyes measured as level (P < 0.01) or C4 deposition function (P < 0.01). CONCLUSIONS: Our study demonstrated that mannose-binding lectin is significantly elevated in inflamed human eyes but virtually undetectable in non-inflamed control eyes, suggesting a role in sight-threatening ocular inflammation. PMID- 21631673 TI - Prevention of retinal ganglion cell swelling by systemic brimonidine in a rat experimental glaucoma model. AB - BACKGROUND: The objective of this study was to evaluate the neuroprotective effect of brimonidine on retinal ganglion cells in rats with elevated intraocular pressure and to characterize the subpopulation of cells that can be rescued, as well as assess the effect of this drug on retinal ganglion cell soma size. METHODS: Episcleral vein cauterization was used to increase intraocular pressure for 5 weeks on left eyes, considering right eyes as intrinsic controls in all cases. All the animals were then given weekly intraperitoneal injections, the experimental group receiving brimonidine, and the control group were administered only phosphate-buffered saline. Surviving retinal ganglion cells were quantified and their area and distribution measured by retrograde labelling with fluorogold. RESULTS: Brimonidine administered systemically has a neuroprotective effect on retinal ganglion cells, which is unrelated to its capacity to lower intraocular pressure. It prevents the increase of cell size that is associated with stages prior to cell death. This phenomenon is particularly evident in the zones of the retina most susceptible to the damage caused by glaucoma (middle and periphery). CONCLUSION: This effect of preventing retinal ganglion cell swelling can be considered as a new marker to study neuroprotection from antiglaucomatous drugs in the early stages of neurodegeneration in glaucoma. PMID- 21631674 TI - Attitudes of consultant ophthalmologists in the UK to initial management of glaucoma patients presenting with severe visual field loss: a national survey. AB - BACKGROUND: Recent National Institute of Clinical Excellence guidance suggests primary surgery should be offered to patients presenting with glaucoma with severe visual field loss. We undertook a survey of UK consultant ophthalmologists to determine if this represents current practice and explore attitudes towards managing patients with advanced glaucoma at presentation. DESIGN: Questionnaire evaluation study. PARTICIPANTS: All consultant ophthalmologists currently practicing in the UK. METHODS: A single-page questionnaire was posted to all consultants (n = 910) currently practicing in the UK along with a pre-paid return envelope. A second questionnaire was sent to non-responders (n = 459). MAIN OUTCOME MEASURES: Questionnaire responses. RESULTS: 626 responses were received representing 68.8% of the population surveyed. 152 (24%) volunteered a specialist interest in glaucoma. Consensus opinion for both glaucoma specialists (64.9%) and non-glaucoma specialists (62.4%) was to start with primary medical therapy, most commonly citing surgical risk as the primary reason (23% and 22%, respectively) for this approach. Most felt the highest intraocular pressure measurement during follow up (measured in clinic) was the most important variable for prevention of further visual loss (60% of glaucoma specialists and 55% of non-glaucoma specialists). Eighty-three per cent of all responders suggested they would change their practice if evidence supporting primary surgery as a safe and more effective approach existed. CONCLUSIONS: Recent National Institute of Clinical Excellence guidance does not reflect the current management approach of UK ophthalmologists. The primary concern was related to potential complications of surgery although most practitioners would be willing to change their practice if evidence existed supporting primary surgery in patients presenting with advanced glaucoma. PMID- 21631675 TI - Vitreous cavity haemorrhage post-vitrectomy for diabetic eye disease: the effect of perioperative anticoagulation and antiplatelet agents. AB - BACKGROUND: To evaluate the effect of perioperative anticoagulation and antiplatelet therapy on postoperative vitreous cavity haemorrhage following pars plana vitrectomy for diabetic eye disease. DESIGN: Retrospective chart review. PARTICIPANTS: 139 patients. METHODS: Retrospective collection of demographic, medical, surgical and postoperative data of all patients undergoing vitrectomy for diabetic eye disease at The Royal Victorian Eye and Ear Hospital. MAIN OUTCOME MEASURE: Correlation of the rates of persistent vitreous cavity haemorrhage and anticoagulation or antiplatelet treatment. RESULTS: Sixty-eight of 155 (43.9%) eyes of 139 patients were on anticoagulation or antiplatelet therapy prior to surgery. At the time of surgery, 29 (42.6%) were on therapy. Eight of 29 (27.6%) patients had significant persistent vitreous cavity haemorrhage in the postoperative period, with four (13.8%) requiring secondary surgery. Thirty-nine (57.4%) patients had discontinued therapy prior to surgery. Among these, four (10.3%) had persistent bleeding, of which three (7.7%) required additional surgery. Six of 87 (6.9%) patients not on any anticoagulation/antiplatelet therapy had persistent postoperative vitreous cavity haemorrhage, with none requiring further surgery. Patients on anticoagulation/antiplatelet therapy at the time of surgery were more likely to experience persistent haemorrhage and subsequent reoperation (OR = 4.8, P = 0.0045 and OR = 6.6, P = 0.024, respectively). CONCLUSION: Perioperative continuation of anticoagulation or antiplatelet treatment appears to increase the risk of persistent postoperative vitreous cavity haemorrhage and the necessity for vitreous cavity washout in this diabetic cohort. Appropriate preoperative cessation of treatment appeared to reduce this risk; however, caution must be taken with regard to the systemic risk associated with cessation of therapy. PMID- 21631676 TI - Outcomes of cataract surgery: a population-based developing world study in the Bhaktapur district, Nepal. AB - BACKGROUND: To evaluate the visual outcome after cataract surgery in a population of Nepal. DESIGN: Population-based cross-sectional study. PARTICIPANTS: Forty years and above residing in Bhaktapur district. METHODS: Subjects were selected from 30 clusters using cluster sampling procedure. All underwent a detailed examination at the base hospital, including logarithm of minimal angle of resolution visual acuity, refraction, applanation tonometry, cataract grading, retinal examination and perimetry when indicated. MAIN OUTCOME MEASURES: Visual acuity after cataract surgery. RESULTS: Out of 4003 subjects examined, 151 had undergone cataract surgery. Pseudophakia was present in 142 (94.0%), aphakia in nine (6%). Presenting and best-corrected visual acuity >=6/18 was achieved in 123 (54.4%) and 164 (72.4%) eyes, respectively. Among the pseudophakic eyes, at presentation 122 (57.5%), 72 (33.9%), 18 (8.5%) and after best correction 162 (76.2%), 33 (15.8%), 17 (8.0%) had visual acuity of >=6/18, <6/18->=6/60 and <6/60, respectively. Retinal disease (35.5%), surgical complications (27.4%) and posterior capsular opacification (14.5%) were the principle causes of visual impairment after best correction in all eyes. There was no significant association in visual outcome based on age, sex, literacy and the duration of surgery. CONCLUSION: Correction of refractive errors, preoperative screening of coincidental diseases, reduction in surgical complication rates and monitoring of postoperative follow-up care has to be addressed seriously in order to improve the outcome of cataract surgery to meet standards proposed by the World Health Organization. In the future, longitudinal studies need to be undertaken to provide specific information on the outcomes of cataract surgery in this community. PMID- 21631677 TI - Corneal endotheliitis triggered by cataract surgery in a Chinese patient with cytomegalovirus anterior uveitis. PMID- 21631678 TI - Use of a novel haemostatic agent: ankaferd blood stopper in conjunctival incisions. AB - BACKGROUND: To investigate the haemostatic efficacy and histopathological effects of a new haemostatic agent, ankaferd blood stopper, in a rat conjunctival incision model. METHODS: Twenty Wistar albino rats were divided into two equal groups (A, B). Limbal incisions of 90-120 degrees were performed in both eyes of all rats. In group A, bleeding at the site of incision was controlled by the administration of ankaferd blood stopper to the right eyes and balanced salt solution to the left eyes. In group B, bleeding was controlled by the application of ankaferd blood stopper to the right eyes and cautery to the left eyes. Time to haemostasis was recorded. After a 4-week period, conjunctival vascularity and postoperative adhesion between Tenon's capsule and sclera were assessed. Additionally, eyes were enucleated and evaluated histopathologically. RESULTS: In group A, the mean bleeding times were 15.2 and 66.7 s for right and left eyes, respectively (P = 0.002). In group B, the mean bleeding times were 17.6 and 17.5 s for right and left eyes, respectively (P = 0.939). Cautery was found to cause significantly more adhesion (P = 0.04). Histopathological examination of the conjunctiva and scleral revealed no statistically significant difference between the samples. CONCLUSIONS: Given the ease of use and lack of histopathological side-effects in the conjunctival incision model, ankaferd blood stopper is promising for use in ophthalmic surgery. Ankaferd blood stopper is a potent haemostatic agent. Its use in ophthalmic surgery should be investigated further in a larger cohort of patients and tested in clinical and experimental models. PMID- 21631679 TI - Prevalence and predictors of refractive error in a genetically isolated population: the Norfolk Island Eye Study. AB - BACKGROUND: We aimed to determine the prevalence and associations of refractive error on Norfolk Island. DESIGN: Population-based study on Norfolk Island, South Pacific. PARTICIPANTS: All permanent residents on Norfolk Island aged >= 15 years were invited to participate. METHODS: Patients underwent non-cycloplegic autorefraction, slit-lamp biomicroscope examination and biometry assessment. Only phakic eyes were analysed. MAIN OUTCOME MEASURES: Prevalence and multivariate associations of refractive error and myopia. RESULTS: There were 677 people (645 right phakic eyes, 648 left phakic eyes) aged >= 15 years were included in this study. Mean age of participants was 51.1 (standard deviation 15.7; range 15-81). Three hundred and seventy-six people (55.5%) were female. Adjusted to the 2006 Norfolk Island population, prevalence estimates of refractive error were as follows: myopia (mean spherical equivalent >= -1.0 D) 10.1%, hypermetropia (mean spherical equivalent >= 1.0 D) 36.6%, and astigmatism 17.7%. Significant independent predictors of myopia in the multivariate model were lower age (P < 0.001), longer axial length (P < 0.001), shallower anterior chamber depth (P = 0.031) and increased corneal curvature (P < 0.001). Significant independent predictors of refractive error were increasing age (P < 0.001), male gender (P = 0.009), Pitcairn ancestry (P = 0.041), cataract (P < 0.001), longer axial length (P < 0.001) and decreased corneal curvature (P < 0.001). CONCLUSIONS: The prevalence of myopia on Norfolk Island is lower than on mainland Australia, and the Norfolk Island population demonstrates ethnic differences in the prevalence estimates. Given the significant associations between refractive error and several ocular biometry characteristics, Norfolk Island may be a useful population in which to find the genetic basis of refractive error. PMID- 21631680 TI - Intravitreal ranibizumab for choroidal neovascularization associated with circumscribed choroidal haemangioma. PMID- 21631681 TI - Assessing the effectiveness of medical treatment for glaucoma. PMID- 21631682 TI - Eyelid deformities from pyodermatitis pyostomatitis vegetans. PMID- 21631683 TI - Prevalence of the refractive errors by age and gender: the Mashhad eye study of Iran. AB - BACKGROUND: Refractive errors are a common eye problem. Considering the low number of population-based studies in Iran in this regard, we decided to determine the prevalence rates of myopia and hyperopia in a population in Mashhad, Iran. DESIGN: Cross-sectional population-based study. PARTICIPANTS: Random cluster sampling. Of 4453 selected individuals from the urban population of Mashhad, 70.4% participated. METHODS: Refractive error was determined using manifest (age > 15 years) and cycloplegic refraction (age <= 15 years). Myopia was defined as a spherical equivalent of -0.5 diopter or worse. An spherical equivalent of +0.5 diopter or worse for non-cycloplegic refraction and an spherical equivalent of +2 diopter or worse for cycloplegic refraction was used to define hyperopia. MAIN OUTCOME MEASURES: Prevalence of refractive errors. RESULTS: The prevalence of myopia and hyperopia in individuals <= 15 years old was 3.64% (95% CI: 2.19-5.09) and 27.4% (95% CI: 23.72-31.09), respectively. The same measurements for subjects > 15 years of age was 22.36% (95% CI: 20.06-24.66) and 34.21% (95% CI: 31.57-36.85), respectively. Myopia was found to increase with age in individuals <= 15 years and decrease with age in individuals > 15 years of age. The rate of hyperopia showed a significant increase with age in individuals > 15 years. The prevalence of astigmatism was 25.64% (95% CI: 23.76-27.51). CONCLUSIONS: In children and the elderly, hyperopia is the most prevalent refractive error. After hyperopia, astigmatism is also of importance in older ages. Age is the most important demographic factor associated with different types of refractive errors. PMID- 21631684 TI - Heavy silicone oil effective in macular hole surgery associated with adult vitelliform macular dystrophy. PMID- 21631685 TI - Ability to determine postoperative status of macular hole in gas-filled eyes by spectral-domain optical coherence tomography. AB - BACKGROUND: To determine whether the status of a macular hole can be determined by spectral-domain optical coherence tomography in gas-filled eyes. DESIGN: Prospective study, university hospital setting. PARTICIPANTS: Twenty-six eyes with a macular hole, seven eyes with a macular hole retinal detachment and four eyes with a macular hole and myopic traction maculopathy underwent vitreous surgery. METHODS: The macular area was examined by spectral-domain optical coherence tomography (OCT-4000) on postoperative days 1, 3, 7 and 30. MAIN OUTCOME MEASURES: Detection of macular hole closure. RESULTS: The status of macular hole was able to be assessed in 24 eyes (92%) on postoperative day 1 and 23 were closed. The condition of the macular hole could not be determined in the other two eyes with a lower volume of gas (<=60%). The macular hole of seven eyes could be seen only when the patients looked upward or downward (P = 0.009). The 11 eyes with a macular hole retinal detachment or a myopic traction maculopathy had a posterior staphyloma, and the macular hole was visible in four eyes on day 1 (P = 0.001). When a closure of the macular hole was detected on day 1, the macular hole remained closed in all of these eyes on day 30. CONCLUSIONS: The status of a macular hole can be determined by spectral-domain optical coherence tomography on postoperative day 1 even in gas-filled eyes. The visibility depends on the volume of intravitreal gas and the absence of a posterior staphyloma. PMID- 21631686 TI - Influence of weather conditions on ophthalmic emergency presentations. PMID- 21631687 TI - Bevacizumab: not as good with more adverse reactions? PMID- 21631688 TI - Ammonia oxidation, denitrification and dissimilatory nitrate reduction to ammonium in two US Great Basin hot springs with abundant ammonia-oxidizing archaea. AB - Many thermophiles catalyse free energy-yielding redox reactions involving nitrogenous compounds; however, little is known about these processes in natural thermal environments. Rates of ammonia oxidation, denitrification and dissimilatory nitrate reduction to ammonium (DNRA) were measured in source water and sediments of two ~ 80 degrees C springs in the US Great Basin. Ammonia oxidation and denitrification occurred mainly in sediments. Ammonia oxidation rates measured using (15)N-NO(3)(-) pool dilution ranged from 5.5 +/- 0.8 to 8.6 +/- 0.9 nmol N g(-1) h(-1) and were unaffected or only mildly stimulated by amendment with NH(4) Cl. Denitrification rates measured using acetylene block ranged from 15.8 +/- 0.7 to 51 +/- 12 nmol N g(-1) h(-1) and were stimulated by amendment with NO(3)(-) and complex organic compounds. The DNRA rate in one spring sediment measured using an (15)N-NO(3)(-) tracer was 315 +/- 48 nmol N g( 1) h(-1). Both springs harboured distinct planktonic and sediment microbial communities. Close relatives of the autotrophic, ammonia-oxidizing archaeon 'Candidatus Nitrosocaldus yellowstonii' represented the most abundant OTU in both spring sediments by 16S rRNA gene pyrotag analysis. Quantitative PCR (qPCR) indicated that 'Ca. N. yellowstonii'amoA and 16S rRNA genes were present at 3.5 3.9 * 10(8) and 6.4-9.0 * 10(8) copies g(-1) sediment. Potential denitrifiers included members of the Aquificales and Thermales. Thermus spp. comprised <1% of 16S rRNA gene pyrotags in both sediments and qPCR for T. thermophilus narG revealed sediment populations of 1.3-1.7 * 10(6) copies g(-1) sediment. These data indicate a highly active nitrogen cycle (N-cycle) in these springs and suggest that ammonia oxidation may be a major source of energy fuelling primary production. PMID- 21631689 TI - Genetic differentiation, recombination and clonal expansion in environmental populations of Cryptococcus gattii in India. AB - Cryptococcus gattii is a ubiquitous eukaryotic pathogen capable of causing life threatening infections in a wide variety of hosts, including both immunocompromised and immunocompetent humans. Since infections by C. gattii are predominantly obtained from environmental exposures, understanding environmental populations of this pathogen is critical, especially in countries like India with a large population and with environmental conditions conducive for the growth of C. gattii. In this study, we analysed 109 isolates of C. gattii obtained from hollows of nine tree species from eight geographic locations in India. Multilocus sequence typing was conducted for all isolates using nine gene fragments. All 109 isolates belonged to the VGI group and were mating type alpha. Population genetic analyses revealed limited evidence of recombination but unambiguous evidence for clonal reproduction and expansion. However, the observed clonal expansion has not obscured the significant genetic differentiation among populations from either different geographic areas or different host tree species. A positive correlation was observed between genetic distance and geographic distance. The results obtained here for environmental populations of C. gattii showed both similarities and differences with those of the closely related Cryptococcus neoformans var. grubii from similar locations and host tree species in India. PMID- 21631690 TI - Low-diversity bacterial community in the gut of the fruitfly Drosophila melanogaster. AB - The bacteria in the fruitfly Drosophila melanogaster of different life stages was quantified by 454 pyrosequencing of 16S rRNA gene amplicons. The sequence reads were dominated by 5 operational taxonomic units (OTUs) at <= 97% sequence identity that could be assigned to Acetobacter pomorum, A. tropicalis, Lactobacillus brevis, L. fructivorans and L. plantarum. The saturated rarefaction curves and species richness indices indicated that the sampling (85,000-159,000 reads per sample) was comprehensive. Parallel diagnostic PCR assays revealed only minor variation in the complement of the five bacterial species across individual insects and three D. melanogaster strains. Other gut-associated bacteria included 6 OTUs with low %ID to previously reported sequences, raising the possibility that they represent novel taxa within the genera Acetobacter and Lactobacillus. A developmental change in the most abundant species, from L. fructivorans in young adults to A. pomorum in aged adults was identified; changes in gut oxygen tension or immune system function might account for this effect. Host immune responses and disturbance may also contribute to the low bacterial diversity in the Drosophila gut habitat. PMID- 21631691 TI - Optimizing vaccine development. AB - Optimizing the development of modern molecular vaccines requires a complex series of interdisciplinary efforts involving basic scientists, immunologists, molecular biologists, clinical vaccinologists, bioinformaticians and epidemiologists. This review summarizes some of the major issues that must be carefully considered. The intent of the authors is to briefly describe key components of the development process to give the reader an overview of the challenges faced from vaccine concept to vaccine delivery. Every vaccine requires unique features based on the biology of the pathogen, the nature of the disease and the target population for vaccination. This review presents general concepts relevant for the design and development of ideal vaccines protective against diverse pathogens. PMID- 21631692 TI - Reduced delays in time to first HIV consultation after diagnosis in France in the antiretroviral therapy era: the possible role of a free care system. PMID- 21631693 TI - Paediatric European Network for Treatment of AIDS response to 2010 revision of World Health Organization recommendations on 'Antiretroviral therapy for HIV infection in infants and children'. PMID- 21631694 TI - Increased incidence of Leydig cell tumours of the testis in the era of improved imaging techniques. AB - OBJECTIVE: To report an observed high frequency of Leydig cell tumours (LCTs) diagnosed at our centre. PATIENTS AND METHODS: Charts of all patients who underwent surgery for a testicular tumour between 1999 and 2008 at our department were searched and data from patients with LCT were collected. Before surgery all patients underwent ultrasound and complete staging. In all but two patients with LCT an organ-sparing surgery was performed. Surgery was performed under ultrasound or palpation guidance. All patients underwent postoperative follow-up. We retrospectively reviewed surgical technique, histology, epidemiology and outcome in all LCT patients. RESULTS: In the study period, 197 testicular tumours were surgically removed of which 29 were diagnosed as LCT (14.7% of 197; further study group) in 25 patients. Mean age of patients with LCT was 45 years (range 21 68 years). Tumour size ranged from 1.2 to 80 mm (mean 10.23 mm). In two patients (8%) the lesion was palpable whereas incidental diagnosis was made in seven patients (28%). In the remaining patients diagnosis was made by ultrasound performed for testicular pain (six patients, 24%) or during infertility or erectile dysfunction evaluation (10 patients, 40%). Definitive histology reported no malignant histopathological features in all but one patient; this particular patient experienced tumour progression after 2 months and died from advanced disease 1 year later. All other patients are free of disease after a mean follow up of 56 months (range 7-93 months). During this period one patient developed a second LCT on the contralateral side; another patient had a recurrence within the same testicle, but on the opposite pole. Both underwent a subsequent organ sparing tumour resection. CONCLUSION: The percentage of LCT (14.7% of all testicular tumours removed) was significantly higher than expected from the literature. One possible explanation for this phenomenon is the increasing use of better ultrasound technology and the subsequent increased detection of small nodules that have not been found in historical series. Use of 'observation-only' for very small lesions detected at infertility clinics is under debate. PMID- 21631695 TI - Comparison of dutasteride and finasteride for treating benign prostatic hyperplasia: the Enlarged Prostate International Comparator Study (EPICS). AB - OBJECTIVE: * To assess the efficacy and safety of dutasteride compared with finasteride in treating men with symptomatic benign prostatic hyperplasia (BPH) for 12 months. PATIENTS AND METHODS: * The Enlarged Prostate International Comparator Study was a multicentre, randomized, double-blind, 12-month, parallel group study. * Men aged >= 50 years with a clinical diagnosis of BPH received once-daily treatment with dutasteride 0.5 mg (n= 813) or finasteride 5 mg (n= 817). After a 4-week placebo run-in period, patients were randomized to receive dutasteride or finasteride for 48 weeks, followed by an optional 24-month, open label phase, during which patients received dutasteride 0.5 mg once daily. * The primary endpoint was change in prostate volume, and the secondary endpoints included improvement in American Urological Association Symptom Index (AUA-SI) scores, improvement in maximum urinary flow rate (Q(max)) and long-term safety in the 24-month open-label phase. RESULTS: * Both dutasteride and finasteride were effective at reducing prostate volume with no significant difference between the two treatments during the study. * Similar reductions in mean AUA-SI scores and Q(max) were also observed for men in both treatment groups. * A similar percentage of adverse events was experienced by patients of both treatment groups, and no new adverse events were reported in the open-label phase. CONCLUSION: * Dutasteride and finasteride, when administered for 12 months, were similarly effective in reducing prostate volume and improving Q(max) and urinary symptoms associated with BPH in men with an enlarged prostate. PMID- 21631696 TI - Clinical utility of diffusion-weighted magnetic resonance imaging in prostate cancer. AB - What's known on the subject? and What does the study add? This article reviews what is currently known about diffusion weighted MRI (DW-MRI) in prostate cancer. This mini-review concisely summarises, for clinical managing patients with prostate cancer, the clinical utility of diffusion weighted MRI. OBJECTIVE: * To review the clinical utility of diffusion-weighted magnetic resonance imaging (DW MRI) in patients with prostate cancer. MATERIAL AND METHODS: * The current literature on prostate cancer and DW-MRI was reviewed. RESULTS: * DW-MRI can be readily acquired on a modern scanner with a short image acquisition time and no need for i.v. contrast medium. * The image contrast is based on the diffusion of water molecules and thus reflects tissue cellularity. * There is increasing evidence that DW- MRI improves the sensitivity and specificity of prostate cancer detection as well as the identification of tumour aggressiveness. * DW-MRI is also showing substantial promise as a response biomarker for both local and metastatic disease CONCLUSIONS: * DW-MRI is proving to be a useful adjunct to conventional T2-weighted MRI sequences. * The eventual role of DW-MRI in combination with other MRI techniques for multiparametric assessment of prostate cancer needs to be defined further. PMID- 21631697 TI - Discussing the sexual consequences of treatment in radiotherapy and urology consultations with couples affected by prostate cancer. AB - OBJECTIVE: To explore the ways in which prostate cancer treatment-induced sexual changes are presented as viable topics for discussion in urology and radiotherapy clinics. PATIENTS AND METHODS: Ethnographic observations were made of 60 consultations between clinicians, patients and partners in clinical oncology and prostate cancer urology clinics. RESULTS: Sexual functioning was discussed infrequently in both clinic settings. Despite the presence of partners in nearly half of consultations, involvement of the partner tended to be minimal. Overall, discussions of wider psychosexual concerns were marginalised in consultations, and there were limited opportunities for couples to discuss the specific impact of prostate cancer and its treatments on sexual functioning. CONCLUSION: Given the potential burden of symptoms and side-effects, there is a need to include discussions of sexual recovery and rehabilitation in consultations, and to provide opportunities to discuss the sexual consequences of treatment with men and their partners. PMID- 21631698 TI - Combining imaging and ureteroscopy variables in a preoperative multivariable model for prediction of muscle-invasive and non-organ confined disease in patients with upper tract urothelial carcinoma. AB - OBJECTIVE: To create a preoperative multivariable model to identify patients at risk of muscle-invasive (pT2+) upper tract urothelial carcinoma (UTUC) and/or non organ confined (pT3+ or N+) UTUC (NOC-UTUC) who potentially could benefit from radical nephroureterectomy (RNU), neoadjuvant chemotherapy and/or an extended lymph node dissection. PATIENTS AND METHODS: We retrospectively analysed data from 324 consecutive patients treated with RNU between 1995 and 2008 at a tertiary cancer centre. Patients with muscle-invasive bladder cancer were excluded, resulting in 274 patients for analysis. Logistic regression models were used to predict pT2+ and NOC-UTUC. Pre-specified predictors included local invasion (i.e. parenchymal, renal sinus fat, or periureteric) on imaging, hydronephrosis on imaging, high-grade tumours on ureteroscopy, and tumour location on ureteroscopy. Predictive accuracy was measured by the area under the curve (AUC). RESULTS: The median follow-up for patients without disease recurrence or death was 4.2 years. Overall, 49% of the patients had pT2+, and 30% had NOC-UTUC at the time of RNU. In the multivariable analysis, only local invasion on imaging and ureteroscopy high grade were significantly associated with pathological stage. AUC to predict pT2+ and NOC-UTUC were 0.71 and 0.70, respectively. CONCLUSIONS: We designed a preoperative prediction model for pT2+ and NOC-UTUC, based on readily available imaging and ureteroscopic grade. Further research is needed to determine whether use of this prediction model to select patients for conservative management vs RNU, neoadjuvant chemotherapy, and/or extended lymphadenectomy will improve patient outcomes. PMID- 21631699 TI - Intermediate-term oncological efficacy of laparoscopic retroperitoneal lymph node dissection for non-seminomatous germ cell testicular cancer. AB - OBJECTIVE: To assess the therapeutic efficacy of laparoscopic retroperitoneal lymph node dissection (L-RPLND) for testicular cancer in patients with nodal disease managed without adjuvant chemotherapy. PATIENTS AND METHODS: Consecutive patients undergoing RPLND were treated laparoscopically. Medical records for 15 patients with pathological stage I and II were reviewed. A modified template dissection was performed laparoscopically. When metastatic disease was noted on intraoperative frozen section, a bilateral template dissection was performed. RESULTS: All patients had predominantly embryonal carcinoma and/or lymphovascular invasion in their orchidectomy specimen. All patients had normal tumour markers after orchidectomy. Laparoscopic RPLND was performed without intraoperative complications. The mean operative time was 299 min and mean length of hospital stay was 1.5 days. After L-RPLND, two patients were pN1 and five patients were pN2. Of the patients with nodal disease, five (two pN1 and three pN2) were followed without chemotherapy for a mean of 30 months with no evidence of recurrence. Isolated pulmonary recurrence occurred in two patients with pathologic stage I disease, and another stage I patient had recurrence in the lung and retroperitoneum outside the dissection template. CONCLUSIONS: Laparoscopic RPLND appears to be safe while providing the benefits of minimally invasive surgery. Although the therapeutic benefit of L-RPLND needs to be confirmed in additional patients and with longer follow-up, our results suggest that L-RPLND provides both diagnostic and therapeutic benefits. PMID- 21631700 TI - Meeting the demand for crop production: the challenge of yield decline in crops grown in short rotations. AB - There is a trend world-wide to grow crops in short rotation or in monoculture, particularly in conventional agriculture. This practice is becoming more prevalent due to a range of factors including economic market trends, technological advances, government incentives, and retailer and consumer demands. Land-use intensity will have to increase further in future in order to meet the demands of growing crops for both bioenergy and food production, and long rotations may not be considered viable or practical. However, evidence indicates that crops grown in short rotations or monoculture often suffer from yield decline compared to those grown in longer rotations or for the first time. Numerous factors have been hypothesised as contributing to yield decline, including biotic factors such as plant pathogens, deleterious rhizosphere microorganisms, mycorrhizas acting as pathogens, and allelopathy or autotoxicity of the crop, as well as abiotic factors such as land management practices and nutrient availability. In many cases, soil microorganisms have been implicated either directly or indirectly in yield decline. Although individual factors may be responsible for yield decline in some cases, it is more likely that combinations of factors interact to cause the problem. However, evidence confirming the precise role of these various factors is often lacking in field studies due to the complex nature of cropping systems and the numerous interactions that take place within them. Despite long-term knowledge of the yield-decline phenomenon, there are few tools to counteract it apart from reverting to longer crop rotations or break crops. Alternative cropping and management practices such as double-cropping or inter-cropping, tillage and organic amendments may prove valuable for combating some of the negative effects seen when crops are grown in short rotation. Plant breeding continues to be important, although this does require a specific breeding target to be identified. This review identifies gaps in our understanding of yield decline, particularly with respect to the complex interactions occurring between the different components of agro-ecosystems, which may well influence food security in the 21(st) Century. PMID- 21631701 TI - Misconduct in scientific publishing. PMID- 21631702 TI - Blood transfusion methods: time to reevaluate? PMID- 21631703 TI - Reassessment Campaign on Veterinary Resuscitation (RECOVER). PMID- 21631704 TI - Application of impedance threshold devices during cardiopulmonary cerebral resuscitation. AB - OBJECTIVE: To review the use of impedance threshold devices (ITD) during CPCR, their proposed mechanism of action, and their application in veterinary medicine. DATA SOURCES: Data sources include scientific reviews and original research publications using the PubMed search engine with the following keywords: 'impedance threshold device' and 'resuscitation' and the Veterinary Information Network search function using the keywords 'impedance threshold device.' HUMAN DATA SYNTHESIS: Studies in human medicine have demonstrated that the use of an ITD during CPCR in patients during out-of-hospital cardiac arrest improves coronary perfusion pressure and cerebral perfusion pressure. This improvement in vital organ blood flow results in increased cardiac output and faster return of spontaneous circulation. The use of an ITD has been studied in people and currently holds a class IIb level of recommendation according to the 2010 American Heart Association Guidelines for CPR and Emergency Cardiovascular Care. This device is recommended as a way to improve hemodynamics during CPCR by enhancing venous return and avoiding hyperventilation, thereby increasing the likelihood of a successful resuscitation. VETERINARY DATA SYNTHESIS: Multiple controlled studies using pigs with ventricular fibrillation induced cardiopulmonary arrest have demonstrated increased myocardial and cerebral perfusion with the use of an ITD. These studies have emphasized the importance of decreasing intrathoracic pressures during the decompression phase of CPCR and avoiding hyperventilation in order to maximize vital organ blood flow. CONCLUSIONS: Use of an ITD during CPCR in human and animal studies has demonstrated improved vital organ perfusion and faster return of spontaneous circulation. However, the majority of these studies have been carried out in people during out-of-hospital cardiac arrest and ventricular fibrillation cardiopulmonary arrest pig models. Further studies evaluating the use of an ITD during CPCR in the veterinary hospital setting are warranted. PMID- 21631705 TI - Myasthenia gravis in dogs with an emphasis on treatment and critical care management. AB - OBJECTIVE: To review the human and veterinary literature on the pathophysiology of myasthenia gravis (MG) and describe treatment options for clinical use in people and animals. DATA SOURCES: Human and veterinary clinical reports, studies and reviews, textbooks, and recent research findings in MG from 1996 present, with a focus on treatment and patient management. HUMAN DATA SYNTHESIS: MG is a well-described condition in people with new research and treatment options available. Many of the newest therapeutic options available in veterinary medicine for MG are based on current strategies used in people with this condition. Seronegative MG is well described in people and provides insight to clinical cases encountered in veterinary medicine when the index of suspicion is high though serologic tests are negative. VETERINARY DATA SYNTHESIS: Previous studies in veterinary medicine focused on the use of acetylcholinesterase inhibitors as the main form of treatment in canine MG. Recent studies, mainly case series and case reports, emphasize the use of immunomodulatory treatments as an alternative for long-term treatment. However, there are no randomized, controlled studies on treatment with immunomodulatory therapy for MG in dogs available to assess the efficacy of this treatment strategy. CONCLUSIONS: Although early recognition of clinical signs is most important in the outcome of patients with MG, further understanding the pathophysiology of MG may lead to earlier diagnosis and novel treatment strategies. The discovery of additional autoantibodies against striated muscle proteins in dogs, should enhance our understanding of diseases affecting the neuromuscular junction. In addition, clinical data for canine MG could be applied to other autoimmune disorders. PMID- 21631706 TI - Influence of transfusion technique on survival of autologous red blood cells in the dog. AB - OBJECTIVE: To determine the effect of 3 differing transfusion techniques on survival of autologous canine RBCs. DESIGN: Prospective, blinded study. SETTING: University Teaching Hospital. ANIMALS: Nine healthy dogs. INTERVENTIONS: Three distinct preparations of RBCs, each representing ~1% of red cell mass, were generated for each dog by biotinylation of RBCs at varying biotin densities. Labeled cells were transfused using 3 techniques (gravity, volumetric pump, syringe pump). Serial determinations of red cell survival were carried out by flow-cytometric analysis of RBCs collected at 7-day intervals for 49 days. In vitro analysis of the effect of transfusion methods on RBC integrity and osmotic fragility were carried out in 7/9 dogs. MEASUREMENTS AND MAIN RESULTS: RBCs administered via volumetric and syringe pumps exhibited a marked decrease in short-term probability of survival compared with RBCs delivered by gravity flow. At 24 hours, only 4/8 and 1/7 dogs had surviving cell populations delivered by volumetric and syringe pump, respectively, compared with 8/8 dogs which had surviving cell populations delivered by gravity flow. Circulating half-life of cells surviving at 24 hours after delivery by volumetric pump was not significantly different to that delivered by gravity flow. No significant effect on in vitro RBC integrity or osmotic fragility was detected in relation to transfusion technique. CONCLUSIONS: Delivery of autologous canine RBCs via mechanical delivery systems was associated with a high risk for early loss of transfused cells. PMID- 21631707 TI - Assessment of a point-of-care cardiac troponin I test to differentiate cardiac from noncardiac causes of respiratory distress in dogs. AB - OBJECTIVES: To (1) determine a reference interval for cardiac troponin I (cTnI) using a point-of-care device in normal dogs and compare the results with those published by the manufacturer and (2) determine if cTnI differs among dogs with cardiogenic and noncardiogenic respiratory distress. DESIGN: Prospective observational study. SETTING: Emergency and referral veterinary hospital. ANIMALS: Twenty-six clinically normal dogs and 67 dogs in respiratory distress. INTERVENTIONS: All dogs underwent whole blood sampling for cTnI concentrations. MEASUREMENTS AND RESULTS: Normal dogs had a median cTnI concentration of 0.03 ng/mL (range 0-0.11 ng/mL). Thirty-six dogs were diagnosed with noncardiogenic respiratory distress with a median cTnI concentration of 0.14 ng/mL (range 0.01 4.31 ng/mL). Thirty-one dogs were diagnosed with cardiogenic respiratory distress with a median cTnI concentration of 1.74 ng/mL (range 0.05-17.1 ng/mL). A significant difference between cTnI concentrations in normal dogs and dogs with noncardiogenic respiratory distress was not detected. Significant differences in cTnI concentrations were found between normals versus cardiogenic and cardiogenic versus noncardiogenic respiratory distress groups. Significant differences in cTnI concentrations were identified in > 10 when compared with the < 5 and the 5 10 years of age groups. Receiver operating curve analysis identified cTnI concentrations > 1.5 ng/mL as the optimal "cut-off point" having a sensitivity of 78% and specificity of 51.5%. The area under the receiver operating curve was 0.72. Overall test accuracy was 65%. CONCLUSIONS: cTnI concentrations were significantly increased in dogs with cardiogenic respiratory distress versus dogs with noncardiogenic respiratory distress and normal dogs. A significant difference between normal dogs and dogs with noncardiogenic causes of respiratory distress was detected. Although highly sensitive when cTnI concentrations exceed 1.5 ng/mL, the test has low specificity. Assessment of cTnI by the methodology used cannot be recommended as the sole diagnostic modality for evaluating the cause of respiratory distress in dogs. PMID- 21631708 TI - Indication, management, and outcome of brachycephalic dogs requiring mechanical ventilation. AB - OBJECTIVES: To evaluate the frequency, and need for mechanical ventilation (MV) in a population of brachycephalic dogs (BD) compared with non-BD. Also, to describe the pre-MV abnormalities, ventilator settings used, the cardiovascular and pulmonary monitoring results and complications encountered in the same BD population. In addition, we sought to identify factors associated with successful weaning and describe outcomes of BD requiring MV. DESIGN: Retrospective observational study (1990-2008). SETTING: University Small Animal Teaching Hospital. ANIMALS: Fifteen BD managed with MV. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Signalment, indication for MV, ventilator settings, arterial blood gas values, duration of MV, complications, and outcome were recorded for each patient enrolled in study. BD were more likely to receive MV than non-BD (P=0.036). Out of the 15 dogs that fulfilled the inclusion criteria 7 (47%) underwent MV for impending respiratory fatigue, 6 (40%) for hypoxemia and 2 for hypercapnea. The most common underlying disease was aspiration pneumonia. Duration of MV ranged from 2 to 240 hours (median 15 hours). Seven patients were weaned (47%). Seven dogs had a temporary tracheostomy tube and 5 of them (71%) were weaned. Dogs that were weaned had a significantly greater preweaning trial PaO2/FiO2 ratio than those that were not (359 +/- 92 versus 210 +/- 57 mm Hg, P=0.025). No significant difference for weaning success between dogs with and those without a tracheostomy was detected (P=0.132). The discharge rate was 27% (all from the respiratory fatigue group). CONCLUSION: Among all dogs admitted to ICU, BD were more likely to receive MV than non-BD. Aspiration pneumonia was frequently identified as the underlying cause of respiratory compromise. The survival rate for BD undergoing MV was not markedly different from previous studies. Weaning of BD from MV may be facilitated by employing preemptive strategies such as performing tracheostomy tube placements. PMID- 21631709 TI - Comparison of whole blood and plasma colloid osmotic pressure in healthy dogs. AB - OBJECTIVE: To determine the difference between colloid osmotic pressure (COP) values determined from plasma versus those determined from whole blood. DESIGN: Prospective observational study. SETTINGS: University veterinary teaching hospital. ANIMALS: Fifty-three healthy dogs. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Whole blood and plasma COP, CBC, plasma biochemistry. In all dogs, plasma COP values were significantly lower (P=0.02) than whole blood COP, with a mean of difference of 0.5 mm Hg. The mean and median whole blood COP was 21.75 and 21.4 mm Hg, respectively, with a range of 17.9-27.1 mm Hg. The mean and median plasma COP was 21.2 and 20.9 mm Hg with a range of 16.7-28.9 mm Hg. CONCLUSIONS: While significant difference exists between plasma and whole blood COP, the individual values are within expected reference intervals for dogs (21 25 mm Hg). Using either sample appears to provide the same information in clinically healthy dog; however, it is recommended that clinicians utilize the same sample type for comparison in an individual patient. PMID- 21631710 TI - Clinical evaluation of serum alcohol dehydrogenase activity in horses with acute intestinal obstruction. AB - OBJECTIVES: To measure serum alcohol dehydrogenase (ADH) activity in horses with acute intestinal obstruction and to determine the diagnostic and prognostic utility of this analyte. DESIGN: Prospective observational study. SETTING: University Veterinary Hospital. ANIMALS: Thirty healthy horses (control group) and 77 horses with acute intestinal obstruction, including 36 horses with nonstrangulating obstruction (23 with left ventral colon impaction and 13 with left dorsal displacement [G1], 22 with small intestinal strangulation [G2], and 19 with colon torsion [G3]). INTERVENTIONS: Serum ADH activity was assayed spectrophotometerically in all horses. Serum lactate concentration and hepatic enzyme (aspartate aminotransferase, gamma-glutamyl transferase, glutamate dehydrogenase) activities were measured using an automatic analyzer. MEASUREMENTS AND MAIN RESULTS: The median [interquartile range] serum ADH activity in healthy horses was 10.5 [8.7-11 U/L]. ADH activity was significantly increased (P<0.05) in G1=16.5 [13.8-18 U/L], G2=40 [20-74.9 U/L], and G3=63.2 [40-78 U/L] compared with healthy controls. Aspartate aminotransferase and glutamate dehydrogenase activities were also significantly increased in G3 in comparison with controls. ADH activity was correlated with serum lactate concentration in G1 and G3, respectively (P<0.01, r=0.55 and 0.8). Other liver enzymes did not show any significant correlation with lactate. ADH activity was directly related to the probability of strangulation; odds ratio=1.11. ADH activity >20 U/L had 80.6% specificity and 80.5% sensitivity for discriminating horses with strangulating obstruction. Twelve horses euthanized before surgery were excluded from the outcome analysis. Increasing ADH activity was associated with nonsurvival; odds ratio=1.03. ADH activity <80 U/L had 94.44% specificity and 66.67% sensitivity for survival. CONCLUSION: Serum ADH activity may be a useful clinical parameter in detecting intestinal strangulation in horses and may provide some prognostic value in horses with acute intestinal obstruction. PMID- 21631711 TI - Acute aldicarb toxicity in dogs: 15 cases (2001-2009). AB - OBJECTIVE: To describe the common clinical signs, laboratory abnormalities, treatment, and prognosis associated with acute aldicarb toxicosis in dogs. DESIGN: Retrospective observational study from 2001 to 2009. SETTING: Urban referral hospital. ANIMALS: Fifteen client-owned dogs. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The most common clinical signs associated with acute aldicarb toxicosis were vomiting, ptyalism, diarrhea, and tremors. Of the 15 dogs, 11 were admitted to the hospital for treatment, 2 were euthanized at presentation and 2 were discharged against medical advice following minimal treatment and lost to follow-up. Laboratory abnormalities included lactic acidosis and hyperglycemia in 12 and 9 patients, respectively. Treatment of hospitalized dogs included induction of emesis with apomorphine (4 dogs), activated charcoal (5), IV fluids (11), atropine (7), methocarbamol (3), diazepam (1), pralidoxime (1) and diphenhydramine (1). Ten of 11 hospitalized dogs survived to discharge; 1 was euthanized following a respiratory arrest after 36 hours of hospitalization. One patient received mechanical ventilation and treatment for pneumonia before discharge from the hospital. The median duration of hospitalization was 22 hours (range 12-168 h). CONCLUSIONS: Acute aldicarb toxicosis carries a good prognosis for survival and hospital discharge with treatment. Supportive care should be considered for at least 18-24 hours to monitor for response to therapy and development of respiratory failure. PMID- 21631712 TI - Retrospective evaluation of postoperative nasotracheal tubes for oxygen supplementation in dogs following surgery for brachycephalic syndrome: 36 cases (2003-2007). AB - OBJECTIVE: To assess the utility of nasotracheal tubes in postoperative oxygen supplementation in dogs following corrective surgery for brachycephalic syndrome. DESIGN: Retrospective study 2003-2007. SETTING: University teaching hospital. ANIMALS: Thirty-six client-owned dogs that underwent corrective surgery for brachycephalic syndrome. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Medical records were reviewed for animals that underwent surgical interventions for brachycephalic syndrome including palatoplasty, ventriculectomy, and rhinoplasty. Data collected included signalment, presenting complaints, analgesic and surgical interventions, type of supplemental oxygen therapy, complications and mortality occurring during hospitalization. A nasotracheal tube (NTT) was placed in 20 dogs at the end of surgery; 16 dogs received other forms of oxygen supplementation (8) or no oxygen supplementation (8) during recovery. The total number of postoperative complications was similar in both groups (8/20 dogs with NTTs and 7/16 in those without NTTs). However, respiratory distress was observed in 5 dogs without NTTs but was not observed in any dog while an NTT was in place. One dog in each group died postoperatively. CONCLUSION: Placement of an NTT was found to be easy and may offer benefit in dogs with brachycephalic syndrome as a noninvasive means of delivering oxygen. The use of NTT may minimize severe postoperative morbidity, in particular by reducing postoperative respiratory distress. PMID- 21631713 TI - 4-Methylpyrazole as a treatment in naturally occurring ethylene glycol intoxication in cats. AB - OBJECTIVE: To describe the clinical experience and therapeutic use of fomepizole (4-methylpyrazole [4-MP]) in 3 cats with naturally occurring ethylene glycol (EG) toxicity. CASE OR SERIES SUMMARY: All cats were documented to be EG positive by an ethylene glycol test kit. This report describes the dose of 4-MP used, available clinicopathological data, and clinical progression during hospitalization. All patients survived to discharge. NEW OR UNIQUE INFORMATION PROVIDED: IV use of 4-MP at 125 mg/kg as an initial dose and 31.25 mg/kg at 12, 24, and 36 hours is safe and effective for treatment of naturally occurring EG toxicity in cats. Increased HCO3 concentrations were noted after IV use of 4-MP. This is the first report documenting the successful treatment of naturally occurring EG intoxication in cats with 4-MP. PMID- 21631714 TI - Presumed immune-mediated hemolytic anemia in two foals with Rhodococcus equi infection. AB - OBJECTIVE: To describe the clinical presentation, case management, and outcome in 2 foals with Rhodococcus equi infection associated with presumptive severe immune mediated hemolytic anemia. SERIES SUMMARY: Two foals diagnosed with R. equi pneumonia on the basis of tracheal wash cultures, thoracic radiographs, and thoracic ultrasonography were concurrently diagnosed with hemolytic anemia. Both foals required whole blood transfusions, and were treated with the antimicrobial combination of rifampin and a macrolide (eg, clarithromycin, erythromycin, or azithromycin). Dexamethasone was used to prevent further hemolysis in both foals, and to treat acute lung injury/acute respiratory distress syndrome in 1 of the foals. Both foals survived, and required prolonged antimicrobial therapy. NEW OR UNIQUE INFORMATION PROVIDED: Although extra-pulmonary disorders are commonly diagnosed in foals infected with R. equi, hemolytic anemia is rarely described. Dexamethasone is considered the treatment of choice for immune-mediated hemolytic anemia, but may be contra-indicated in foals with severe bacterial infections. In these foals, a relatively low dose and short duration of dexamethasone was utilized in an attempt to minimize immune suppression, although early discontinuation in 1 foal precipitated a second hemolytic crisis. PMID- 21631715 TI - Uveitis associated with septic peritonitis in a cat. AB - OBJECTIVE: To report a case of bilateral uveitis believed to be a consequence of septic peritonitis in a 19-month-old cat. CASE SUMMARY: Bilateral anterior uveitis with suspicion of extension to the posterior segment was documented in a previously healthy young cat during hospitalization for severe septic peritonitis. Based on medical history and other findings uveitis was believed to result from concurrent abdominal sepsis, due either to metastatic seeding of bacterial organisms or to effects of bacterial toxins and inflammatory mediators on the blood-aqueous barrier. The cat was surgically and medically managed, and made a full recovery with respect to both his ocular and his abdominal disease. NEW OR UNIQUE INFORMATION PROVIDED: Ocular complications secondary to systemic sepsis are well documented in people but seldom reported in the veterinary literature. To the authors' knowledge this is the first report of uveitis linked to septic peritonitis in any veterinary species and the first to report sepsis related uveitis in a cat. Ocular inflammatory disease in the context of critical illness deserves attention as a potential significant source of morbidity. The development of ocular inflammatory disease may serve as a sentinel lesion for systemic sepsis and other life-threatening conditions. PMID- 21631723 TI - Pregnancy in dialysis-dependent women--the importance of frequent dialysis and collaborative care: a case report. AB - An increasing number of successful pregnancies have been reported among women on chronic hemodialysis. Even with reduced fertility and high risk of complications, women of childbearing age receiving hemodialysis, should not be discouraged from pregnancy. Practitioners should be familiar with the effects of renal disease on pregnancy, consult patients about the possibility of pregnancy and its hazards and provide, if necessary, prompt surveillance and treatment. This paper describes the case of an unplanned but successful pregnancy of a woman receiving hemodialysis, emphasizing pregnancy management, mother's response evaluation, and infant growth. PMID- 21631725 TI - Delayed removal of a primary incisor embedded in the upper lip after dental trauma: a case report about the importance of soft tissue examination. AB - This article describes a child patient who initially had inadequate treatment and suffered concomitant soft tissue trauma involving complete displacement of a primary central incisor into the lip tissue. The primary tooth was subsequently removed by surgery under general anesthesia. Despite the delay in diagnosis, there was an excellent outcome following removal of the embedded tooth. This paper again emphasizes the importance of an accurate history, physical, and radiographic evaluation of these patients in the acute phase. The importance of soft tissue inspection even in cases that are presented late for dental trauma management is also highlighted. This case shows that educated emergency room staff preferably including an oral and maxillofacial surgeon is required for a proper emergency management in orofacial traumas. PMID- 21631726 TI - Fiber Bragg grating sensor for measurement of impact absorption capability of mouthguards. AB - BACKGROUND: There is no standard technique to monitor impact absorption capability of mouthguards. Earlier investigations have established that strain transferred to the teeth through mouthguard is a good indication of their efficiency. In the present study, a unique experimental scheme utilizing fiber Bragg gratings (FBGs) as distributed strain sensors is proposed and investigated to estimate impact absorption capability of custom-made mouthguard. The proposed methodology is useful due to advantages such as, very small size and flexibility for ease of bonding, self-referencing, and multiplexing capability of using FBG sensors. MATERIAL AND METHODS: Finite-element analysis was performed to simulate the stress distribution due to impact on the mouthguard. The FBGs were fabricated by exposing the core of photosensitive fiber to intense Ultra-Violet light through a 'phase mask'. One FBG sensor was bonded on the jaw model and another on the mouthguard surface at similar positions, so that both gratings are simultaneously affected by impact. Two different sets of the sensors were used, one for the anterior region and another for posterior region. The impact was produced using customized pendulum device with interchangeable impact objects i.e. cricket ball, hockey ball, and steel ball. Response of gratings was monitored using optical spectrum analyzer and strain induced due to each impact was determined from the Bragg wavelength shifts for each grating. RESULTS AND CONCLUSIONS: Strain induced due to impact was calculated from the Bragg wavelength shifts. Difference in the strain values for the two gratings is interpreted as impact energy absorbed by the mouthguard. The Bragg wavelength shifts (induced strain) for FBG bonded on the jaw model was much lower than the shift for FBG bonded on the mouthguard, indicating that most of the impact energy is absorbed by the mouthguard. PMID- 21631727 TI - Environmental stress alters genetic regulation of novelty seeking in vervet monkeys. AB - Considerable attention has been paid to identifying genetic influences and gene environment interactions that increase vulnerability to environmental stressors, with promising but inconsistent results. A nonhuman primate model is presented here that allows assessment of genetic influences in response to a stressful life event for a behavioural trait with relevance for psychopathology. Genetic and environmental influences on free-choice novelty seeking behaviour were assessed in a pedigreed colony of vervet monkeys before and after relocation from a low stress to a higher stress environment. Heritability of novelty seeking scores, and genetic correlations within and between environments were conducted using variance components analysis. The results showed that novelty seeking was markedly inhibited in the higher stress environment, with effects persisting across a 2-year period for adults but not for juveniles. There were significant genetic contributions to novelty seeking scores in each year (h(2) = 0.35-0.43), with high genetic correlations within each environment (rhoG > 0.80) and a lower genetic correlation (rhoG = 0.35, non-significant) between environments. There were also significant genetic contributions to individual change scores from before to after the move (h(2) = 0.48). These results indicate that genetic regulation of novelty seeking was modified by the level of environmental stress, and they support a role for gene-environment interactions in a behavioural trait with relevance for mental health. PMID- 21631728 TI - Adenosine diphosphate ribosylation factor-GTPase-activating protein stimulates the transport of AUX1 endosome, which relies on actin cytoskeletal organization in rice root development. AB - Polar auxin transport, which depends on polarized subcellular distribution of AUXIN RESISTANT 1/LIKE AUX1 (AUX1/LAX) influx carriers and PIN-FORMED (PIN) efflux carriers, mediates various processes of plant growth and development. Endosomal recycling of PIN1 is mediated by an adenosine diphosphate (ADP)ribosylation factor (ARF)-GTPase exchange factor protein, GNOM. However, the mediation of auxin influx carrier recycling is poorly understood. Here, we report that overexpression of OsAGAP, an ARF-GTPase-activating protein in rice, stimulates vesicle transport from the plasma membrane to the Golgi apparatus in protoplasts and transgenic plants and induces the accumulation of early endosomes and AUX1. AUX1 endosomes could partially colocalize with FM4-64 labeled early endosome after actin disruption. Furthermore, OsAGAP is involved in actin cytoskeletal organization, and its overexpression tends to reduce the thickness and bundling of actin filaments. Fluorescence recovery after photobleaching analysis revealed exocytosis of the AUX1 recycling endosome was not affected in the OsAGAP overexpression cells, and was only slightly promoted when the actin filaments were completely disrupted by Lat B. Thus, we propose that AUX1 accumulation in the OsAGAP overexpression and actin disrupted cells may be due to the fact that endocytosis of the auxin influx carrier AUX1 early endosome was greatly promoted by actin cytoskeleton disruption. PMID- 21631729 TI - One women's voice for all. PMID- 21631731 TI - Current approach to the man with acute phase Peyronie's disease--case presentation and discussion. PMID- 21631734 TI - 2010 Sandra Leiblum Presidential Address. PMID- 21631741 TI - Evidence-based healthcare. PMID- 21631735 TI - Sex therapy and combined (sex and medical) therapy. PMID- 21631742 TI - Effectiveness of nurse-led cardiac clinics in adult patients with a diagnosis of coronary heart disease. AB - Background Coronary heart disease is the major cause of illness and death in Western countries and this is likely to increase as the average age of the population rises. Consumers with established coronary heart disease are at the highest risk of experiencing further coronary events. Lifestyle measures can contribute significantly to a reduction in cardiovascular mortality in established coronary heart disease. Improved management of cardiac risk factors by providing education and referrals as required has been suggested as one way of maintaining quality care in patients with established coronary heart disease. There is a need to ascertain whether or not nurse-led clinics would be an effective adjunct for patients with coronary heart disease to supplement general practitioner advice and care. Objectives The objective of this review was to present the best available evidence related to nurse-led cardiac clinics. Inclusion criteria This review considered any randomised controlled trials that evaluated cardiac nurse-led clinics. In the absence of randomised controlled trials, other research designs such as non-randomised controlled trials and before and after studies were considered for inclusion. Participants were adults (18 years and older) with new or existing coronary heart disease. The interventions of interest to the review included education, assessment, consultation, referral and administrative structures. Outcomes measured included adverse event rates, readmissions, admissions, clinical and cost effectiveness, consumer satisfaction and compliance with therapy. Results Based on the search terms used, 80 papers were initially identified and reviewed for inclusion; full reports of 24 of these papers were retrieved. There were no papers included that addressed cost effectiveness or adverse events; and none addressed the outcome of referrals. A critical appraisal of the 24 remaining papers identified a total of six randomised controlled trials that met the inclusion criteria. Two studies addressed nurse-led clinics for patients diagnosed with angina, one looked at medication administration and the other looked at educational plans. A further four studies compared secondary preventative care with a nurse-led clinic and general practitioner clinic. One specifically compared usual care versus shared care introduced by nurses for patients awaiting coronary artery bypass grafting. Of the remaining three studies, two have been combined in the results section, as they are an interim report and a final report of the same study. Because of inconsistencies in reporting styles and outcome measurements, meta-analysis could not be performed on all outcomes. However, a narrative summary of each study and comparisons of specific outcomes assessed from within each study has been developed. Although not all outcomes obtained statistical significance, nurse-led clinics were at least as effective as general practitioner clinics for most outcomes. Recommendations The following recommendations are made: * The use of nurse-led clinics is recommended for patients with coronary heart disease (Level II). * Utilise nurse-led clinics to increase clinic attendance and follow-up rates (Level II). * Nurse-led clinics are recommended for patients who require lifestyle changes to decrease their risk of adverse outcomes associated with coronary heart disease (Level II). PMID- 21631743 TI - Effectiveness of primary conservative management for infants with obstetric brachial plexus palsy. AB - Background Obstetric brachial plexus palsy, a complication of childbirth, occurs in 1-3 per 1000 live births internationally. Traction and/or compression of the brachial plexus is thought to be the primary mechanism of injury and this may occur in utero, during the descent through the birth canal or during delivery. This results in a spectrum of injuries that vary in severity, extent of damage and functional use of the affected upper limb. Most infants receive treatment, such as conservative management (physiotherapy, occupational therapy) or surgery; however, there is controversy regarding the most appropriate form of management. To date, no synthesised evidence is available regarding the effectiveness of primary conservative management for obstetric brachial plexus palsy. Objectives The objective of this review was to systematically assess the literature and present the best available evidence that investigated the effectiveness of primary conservative management for infants with obstetric brachial plexus palsy. Search strategy A systematic literature search was performed using 14 databases: TRIP, MEDLINE, CINAHL, AMED, Web of Science, Proquest 5000, Evidence Based Medicine Reviews, Expanded Academic ASAP, Meditext, Science Direct, Physiotherapy Evidence Database, Proquest Digital Dissertations, Open Archives Initiative Search Engine, Australian Digital Thesis Program. Those studies that were reported in English and published over the last decade (July 1992 to June 2003) were included in this review. Selection criteria Quantitative studies that investigated the effectiveness of primary conservative management for infants with obstetric brachial plexus palsy were eligible for inclusion in this review. This excluded studies that solely investigated the effect of primary surgery for these infants, management of secondary deformities and the investigation of the effects of pharmacological agents, such as botulinum toxin. Data collection and analysis Two independent reviewers assessed the eligibility of each study for inclusion into the review, the study design used and its methodological quality. Where any disagreement occurred, consensus was reached by discussion. Studies were assessed for clinical homogeneity by considering populations, interventions and outcomes. Where heterogeneity was present, synthesis was undertaken in a narrative format. Results Eight studies were included in the review. Most were ranked low on the Hierarchy of Evidence (no randomised controlled trials were found), and had only fair methodological quality. Conservative management was variable and could consist of active or passive exercise, splints or traction. All studies lacked a clear description of what constituted conservative management, which would not allow the treatment to be replicated in the clinical setting. A variety of outcome instruments were used, none of which had evidence of validity, reliability or sensitivity to detect change. Furthermore, less severely affected infants were selected to receive conservative management. Therefore, it is difficult to draw conclusions regarding the effectiveness of conservative management for infants with obstetric brachial plexus palsy. Conclusions There is scant, inconclusive evidence regarding the effectiveness of primary conservative intervention for infants with obstetric brachial plexus palsy. Further research should be directed to develop outcome instruments with sound psychometric properties for infants with obstetric brachial plexus palsy and their families. These outcome instruments should then be used in well designed comparative studies. PMID- 21631744 TI - Knowledge retention from preoperative patient information. AB - Background Preoperative education is a common feature of the preoperative preparation for many surgical procedures. It is anticipated that this education will result in beneficial outcomes for the patient. Many studies have evaluated the effectiveness of different formats used to deliver the information, and the effect of this information on a variety of outcomes. While there has been substantial research and several meta-analyses undertaken on different aspects of preoperative education, there has been no previous attempt to summarise this body of research to evaluate its impact on knowledge and understanding of patients. Objective The objective of this review was to present the best available evidence related to knowledge retention and/or correct performance of postoperative activities after preoperative patient education. Review method This review considered all studies that included adults in a hospital setting, either as inpatients or same day surgical patients, and who received some form of information and/or instruction before an operative procedure. Interventions were the methods of preoperative patient education, instruction or teaching, and included evaluations of the effectiveness of different presentations such as: * written information; * audio-visual aids; * computer-assisted instruction; * learning packages in either group or individual formats, at either pre-admission or post-admission. The primary outcomes were those associated with the understanding of the information related to the operative or postoperative period that were provided in the intervention and included: 1 increased knowledge; 2 ability to perform postoperative activities; 3 time to teach skills. This review considered randomised controlled trials that evaluated forms of preoperative patient education and their effect on patient understanding, knowledge and ability to perform postoperative activities. Results The findings of this review support the use of pamphlets to inform patients and to improve their skills. The role of videos as a preoperative instruction tool has not been rigorously evaluated. However, existing studies support the use of preoperative videos to improve patient knowledge and skill. The data suggest that the instructional method, the act of educating a patient by delivering directions for actions or behaviour is useful for improving patients' knowledge of their treatment and their ability to perform and comply with required exercises. However, instruction is likely to be more effective if provided before admission. If teaching is to be done after admission, using a group format has been shown to be equally as effective as individual instruction. Conclusions Although numerous studies have been performed on many aspects of preoperative education, little high-quality research has assessed the effectiveness of this information on patient knowledge and ability to perform specific skills such as exercises. Further research is required to examine: (i) the effectiveness of pamphlets and other written material for people with English as a second language or limited literary skills; (ii) the accuracy of information provided in preoperative pamphlets; (iii) the effectiveness of pamphlets on general populations; (iv) the role of videos and learning packages; (v) the effectiveness of preoperative instruction; and (vi) the changes in patient misconceptions with the provision of preoperative information. PMID- 21631745 TI - Effectiveness of topical skin care provided in aged care facilities. AB - Background The 2001 Australian census revealed that adults aged 65 years and over constituted 12.6% of the population, up from 12.1% in 1996. It is projected that this figure will rise to 21% or 5.1 million Australians by 2031. In 1998, 6% (134 000) of adults in Australia aged 65 years and over were residing in nursing homes or hostels and this number is also expected to rise. As skin ages, there is a decreased turnover and replacement of epidermal skin cells, a thinning subcutaneous fat layer and a reduced production of protective oils. These changes can affect the normal functions of the skin such as its role as a barrier to irritants and pathogens, temperature and water regulation. Generally, placement in a long-term care facility indicates an inability of the older person to perform all of the activities of daily living such as skin care. Therefore, skin care management protocols should be available to reduce the likelihood of skin irritation and breakdown and ultimately promote comfort of the older person. Objectives The objective of this review was to determine the best available evidence for the effectiveness and safety of topical skin care regimens for older adults residing in long-term aged care facilities. The primary outcome was the incidence of adverse skin conditions with patient satisfaction considered as a secondary outcome. Search strategy A literature search was performed using the following databases: PubMed (NLM) (1966-4/2003), Embase (1966-4/2003), CINAHL (1966-4/2003), Current Contents (1993-4/2003), Cochrane Library (1966-2/2003), Web of Science (1995-12/2002), Science Citation Index Expanded and ProceedingsFirst (1993-12/2002). Health Technology Assessment websites were also searched. No language restrictions were applied. Selection criteria Systematic reviews of randomised controlled trials, randomised and non-randomised controlled trials evaluating any non-medical intervention or program that aimed to maintain or improve the integrity of skin in older adults were considered for inclusion. Participants were 65 years of age or over and residing in an aged care facility, hospital or long-term care in the community. Studies were excluded if they evaluated pressure-relieving techniques for the prevention of skin breakdown. Data collection and analysis Two independent reviewers assessed study eligibility for inclusion. Study design and quality were tabulated and relative risks, odds ratios, mean differences and associated 95% confidence intervals were calculated from individual comparative studies containing count data. Results The resulting evidence of the effectiveness of topical skin care interventions was variable and dependent upon the skin condition outcome being assessed. The strongest evidence for maintenance of skin condition in incontinent patients found that disposable bodyworn incontinence protection reduced the odds of deterioration of skin condition compared with non-disposable bodyworns. The best evidence for non-pressure relieving topical skin care interventions on pressure sore formation found the no-rinse cleanser Clinisan to be more effective than soap and water at maintaining healthy skin (no ulcers) in elderly incontinent patients in long-term care. The quality of studies examining the effectiveness of topical skin care interventions on the incidence of skin tears was very poor and inconclusive. Topical skin care for prevention of dermatitis found that Sudocrem could reduce the redness of skin compared with zinc cream if applied regularly after each pad change, but not the number of lesions. Topical skin care on dry skin found the Bag Bath/Travel Bath no-rinse skin care cleanser to be more effective at preventing overall skin dryness and most specifically flaking and scaling when compared with the traditional soap and water washing method in residents of a long-term care facility. Information on the safety of topical skin care interventions is lacking. Therefore, because of the lack of evidence, no recommendation on the safety on any intervention included in this review can be made. PMID- 21631746 TI - Effectiveness of a pelvic floor muscle exercise program on urinary incontinence following childbirth. AB - EXECUTIVE SUMMARY: Objectives The primary objective of this review was to determine, from the available evidence, the effectiveness of an antenatal and/or a post-natal program of pelvic floor muscle exercises (PFME) compared with usual care on preventing, reducing or resolving the incidence and severity of stress incontinence, urge incontinence or mixed stress and urge urinary incontinence following childbirth. Secondary objectives were included to examine the effectiveness of a PFME program on pelvic floor muscle strength and on encouraging adherence to an exercising program. INCLUSION CRITERIA: TYPES OF STUDIES: Randomised controlled trials and non-randomised controlled trials were included in the review if, in relation to urinary incontinence, and/or adherence to a PFME program, and/or pelvic floor muscle strength, the following had been explored: * antenatal PFME compared with usual care; * post-natal PFME compared with usual care; * a PFME program compared with usual care. Usual care is commonly used to describe the care women normally receive from health professionals in the antenatal and/or post-natal period. In some cases usual care includes a standard information package given to all women attending the health service and in others it is advice about performing PFME. TYPES OF PARTICIPANTS: Participants included women who experienced a spontaneous onset of labour and who subsequently delivered at more than 20 weeks gestation either vaginally, both spontaneous and assisted, or by non-elective caesarean section. EXCLUSIONS: * women who delivered by elective caesarean section; * women experiencing post partum overflow urinary incontinence. TYPES OF INTERVENTIONS: 1 Pelvic floor muscle exercises. 2 PFME instruction and a PFME program's components, such as educational materials, feedback (including biofeedback, e.g. information about strength of pelvic floor muscle contractions by various means) and number of PFME. EXCLUSIONS: * electrical stimulation of pelvic floor muscles; * vaginal cones; or * other adjunct therapies. In studies that included a subgroup treated with one of these interventions, the results of the subgroup were excluded from the review's analysis. TYPES OF OUTCOME MEASURES: Outcomes that were of interest: * non-occurrence of urinary incontinence following childbirth; * a change in the frequency, duration or severity (as appropriate) of urinary incontinence up to 12 months following childbirth. * a change in the strength of pelvic floor muscle contractions; * period of time PFME continued after initial instruction; * frequency of PFME undertaken; * women's awareness of the importance of PFME; * satisfaction with PFME instruction. Search strategy All major electronic sources of information relevant to the topic (e.g. PubMed, CINAHL and the Cochrane Library) were searched to identify published and unpublished studies and previous work in the field. Printed journals were hand-searched and reference lists checked for potentially useful research. The review included any studies undertaken between 1981 and 2003. The search did not attempt to locate unpublished research before 1991. Assessment of quality An independent Review Panel carried out quality assessment of studies. Two members of the panel, using quality assessment checklists developed for the review, reviewed each study. Disagreements between reviewers were resolved through discussion or a third reviewer examining a study. Data extraction and analysis A data extraction tool was developed to extract data relating to participant characteristics, study methods, interventions and outcomes. Two reviewers independently extracted the required data. Randomised controlled trials included in the review were pooled in several meta-analyses using RevMan software program. Heterogeneity between studies was determined to ensure that they were sufficiently similar to allow for the pooling of their results. Non-randomised controlled trials were discussed in narrative comparisons. Results Six randomised controlled trials met the inclusion criteria for the primary objective of the systematic review. The results of this review indicate that antenatal PFME and post-natal PFME are effective in resolving or reducing urinary incontinence following childbirth. There was insufficient evidence to conclude that PFME can prevent urinary incontinence in post-partum women. In most of these studies women were selected randomly and therefore included women without urinary incontinence and women with urinary incontinence. Two randomised controlled trials selected their sample on the criteria of existing post-partum urinary incontinence. A subgroup analysis of these studies showed that post-natal PFME also have a significant effect on reducing or resolving urinary incontinence in women with existing post-partum urinary incontinence. Seven randomised controlled trials and three non-randomised controlled trials met the inclusion criteria for the secondary objectives of the review. Findings of the studies included in the review suggest a PFME program will improve the frequency with which women perform PFME. Two studies found that women receiving the intervention (a PFME program) and who were performing PFME regularly in the month before data collection were significantly less likely to have any incontinence. The review's results support previous findings showing there is little evidence that a high-intensity PFME program is more effective than a low-intensity PFME regimen of exercising. No conclusions about the effectiveness of feedback to a woman about pelvic floor muscle strength, for example, perineometer measures, as part of a PFME program can be reached. The mixed results of this review mean that no conclusions can be reached about the effectiveness of a PFME program, antenatal or post-natal, on improving pelvic floor muscle strength. A number of studies reported a high percentage of women lost to follow-up and the data collected in most of the studies relied on self reports relating to urinary incontinence and/or frequency of exercising. These factors may have affected the overall results of the review. However, wherever possible, tests for heterogeneity were carried out to determine if studies should be combined in meta-analyses and in other cases the results' limitations are acknowledged. Implications for practice In terms of the effectiveness of PFME programs, the results of this review indicate that urinary incontinence following childbirth can be improved by performing PFME and that any form of a specific PFME program appears to improve exercising frequency. However, the value of individual components of PFME programs, such as take-home materials, reminder telephone calls and feedback of exercising effectiveness, is less clear. * Encourage women to undertake both antenatal and post-natal PFME (E1). * Pay particular attention to women with antenatal and post-natal urinary incontinence in providing advice and PFME instruction (E1). * To encourage adherence and continuation, PFME education programs should be multifaceted with a number of components, rather than only supplying an information booklet (E4). * Include PFME as a specific program in all antenatal and post-natal care, incorporating at least two individual instruction sessions into the program (E1). * Provide post partum contact, particularly for those discharged early, either by telephone, electronic or home visits (E4). * Design pelvic floor muscle home exercise programs that are realistic given the demands on a mother and that can be incorporated into her daily routine in terms of number and frequency. Two or more training sessions per week are recommended (E4). * Health professionals working with women in the post-partum period should ask about symptoms of incontinence to ensure assistance is offered to those experiencing urinary incontinence (E4). PMID- 21631747 TI - Association between pacifier use and breast-feeding, sudden infant death syndrome, infection and dental malocclusion. AB - OBJECTIVE: To critically review all literature related to pacifier use for full term healthy infants and young children. The specific review questions addressed are: What is the evidence of adverse and/or positive outcomes of pacifier use in infancy and childhood in relation to each of the following subtopics: * breast feeding; * sudden infant death syndrome; * infection; * dental malocclusion. INCLUSION CRITERIA: Specific criteria were used to determine which studies would be included in the review: (i) the types of participants; (ii) the types of research design; and (iii) the types of outcome measures. To be included a study has to meet all criteria. TYPES OF PARTICIPANTS: The participants included in the review were healthy term infants and healthy children up to the age of 16 years. Studies that focused on preterm infants, and infants and young children with serious illness or congenital malformations were excluded. However, some total population studies did include these children. Types of research design: It became evident early in the review process that very few randomised controlled trials had been conducted. A decision was made to include observational epidemiological designs, specifically prospective cohort studies and, in the case of sudden infant death syndrome research, case-control studies. Purely descriptive and cross-sectional studies were excluded, as were qualitative studies and all other forms of evidence. A number of criteria have been proposed to establish causation in the scientific and medical literature. These key criteria were applied in the review process and are described as follows: (i) consistency and unbiasedness of findings; (ii) strength of association; (iii) temporal sequence; (iv) dose-response relationship; (v) specificity; (vi) coherence with biological background and previous knowledge; (vii) biological plausibility; and (viii) experimental evidence. Studies that did not meet the requirement of appropriate temporal sequencing of events and studies that did not present an estimate of the strength of association were not included in the final review. Types of outcome measures: Our specific interest was pacifier use related to: * breast-feeding; * sudden infant death syndrome; * infection; * dental malocclusion. Studies that examined pacifier use related to procedural pain relief were excluded. Studies that examined the relationship between pacifier use and gastro-oesophageal reflux were also excluded as this information has been recently presented as a systematic review. SEARCH STRATEGY: The review comprised published and unpublished research literature. The search was restricted to reports published in English, Spanish and German. The time period covered research published from January 1960 to October 2003. A protocol developed by New Zealand Health Technology Assessment was used to guide the search process. The search comprised bibliographic databases, citation searching, other evidence based and guidelines sites, government documents, books and reports, professional websites, national associations, hand search, contacting national/international experts and general internet searching. ASSESSMENT OF QUALITY: All studies identified during the database search were assessed for relevance to the review based on the information provided in the title, abstract and descriptor/MeSH terms, and a full report was retrieved for all studies that met the inclusion criteria. Studies identified from reference list searches were assessed for relevance based on the study title. Keywords included: dummy, dummies, pacifier(s), soother(s), comforter(s), non-nutritive sucking, infant, child, infant care. Initially, studies were reviewed for inclusion by pairs of principal investigators. Authorship of articles was not concealed from the reviewers. Next, the methodological quality of included articles was assessed independently by groups of three or more principal investigators and clinicians using a checklist. All 20 studies that were accepted met minimum set criteria, but few passed without some methodological concern. DATA EXTRACTION: To meet the requirements of the Joanna Briggs Institute, reasons for acceptance and non-acceptance at each phase were clearly documented. An assessment protocol and report form was developed for each of the three phases of review. The first form was created to record investigators' evaluations of studies included in the initial review. Those studies that failed to meet strict inclusion criteria were excluded at this point. A second form was designed to facilitate an in-depth critique of epidemiological study methodology. The checklist was pilot tested and adjustments were made before reviewers were trained in its use. When reviewers could not agree on an assessment, it was passed to additional reviewers and discussed until a consensus was reached. At this stage, studies other than cohort, case-control and randomised controlled trials were excluded. Issues of clarification were also addressed at this point. The final phase was that of integration. This phase, undertaken by the principal investigators, was assisted by the production of data extraction tables. Through a process of trial and error, a framework was formulated that adequately summarised the key elements of the studies. This information was tabulated under the following headings: authors/setting, design, exposure/outcome, confounders controlled, analysis and main findings. RESULTS: With regard to the breast-feeding outcome, 10 studies met the inclusion criteria, comprising two randomised controlled trials and eight cohort studies. The research was conducted between 1995 and 2003 in a wide variety of settings involving research participants from diverse socioeconomic and cultural backgrounds. Information regarding exposure and outcome status, and potential confounding factors was obtained from: antenatal and postnatal records; interviews before discharge from obstetric/midwifery care; post-discharge interviews; and post-discharge postal and telephone surveys. Both the level of contact and the frequency of contact with the informant, the child's mother, differed widely. Pacifier use was defined and measured inconsistently, possibly because few studies were initiated expressly to investigate its relationship with breast-feeding. Completeness of follow-up was addressed, but missing data were not uniformly identified and explained. When comparisons were made between participants and non-participants there was some evidence of differential loss and a bias towards families in higher socioeconomic groups. Multivariate analysis was undertaken in the majority of studies, with some including a large number of sociodemographic, obstetric and infant covariates and others including just maternal age and education. As might be expected given the inconsistency of definition and measurement, the relationship between pacifier use and breast feeding was expressed in many different ways and a meta-analysis was not appropriate. In summary, only one study did not report a negative association between pacifier use and breast-feeding duration or exclusivity. Results indicate an increase in risk for a reduced overall duration of breast-feeding from 20% to almost threefold. The data suggest that very infrequent use may not have any overall negative impact on breast-feeding outcomes. Six sudden infant death syndrome case-control studies met the criteria for inclusion. The research was conducted with information gathered between 1984 and 1999 in Norway, UK, New Zealand, the Netherlands and USA. Exposure information was obtained from a variety of sources including: hospital and antenatal records, death scene investigation, and interview and questionnaire. Information for cases was sought within 2 days after death, within 2-4 weeks after death and in one study between 3 and 11 years after death. Information for controls was sought from as early as 4 days of a nominated sudden infant death syndrome case, to between 1 and 7 weeks from the case date, and again in one study some 3-11 years later. In the majority of the studies case ascertainment was determined by post-mortem. Pacifier use was again defined and measured somewhat inconsistently. All studies controlled for confounding factors by matching and/or using multivariate analysis. Generally, antenatal and postnatal factors, as well as infant care practices, and maternal, family and socioeconomic issues were considered. (ABSTRACT TRUNCATED) PMID- 21631748 TI - Nursing and midwifery management of hypoglycaemia in healthy term neonates. AB - EXECUTIVE SUMMARY: OBJECTIVES: The primary objective of this review was to determine the best available evidence for maintenance of euglycaemia* in healthy term neonates, and the management of asymptomatic hypoglycaemia in otherwise healthy term neonates. INCLUSION CRITERIA: TYPES OF STUDIES: The review included any relevant published or unpublished studies undertaken between 1995 and 2004. Studies that focus on the diagnostic accuracy of point-of-care devices for blood glucose screening and/or monitoring in the neonate were initially included as a subgroup of this review. However, the technical nature and complexity of the statistical information published in diagnostic studies retrieved during the literature search stage, as well as the considerable volume of published research in this area, suggested that it would be more feasible to analyse diagnostic studies in a separate systematic review. TYPES OF PARTICIPANTS: The review focused on studies that included healthy term (37- to 42-week gestation) appropriate size for gestational age neonates in the first 72 h after birth. EXCLUSIONS: * preterm or small for gestational age newborns; * term neonates with a diagnosed medical or surgical condition, congenital or otherwise; * babies of diabetic mothers; * neonates with symptomatic hypoglycaemia; * large for gestational age neonates (as significant proportion are of diabetic mothers). TYPES OF INTERVENTION: All interventions that fell within the scope of practice of a midwife/nurse were included: * type (breast or breast milk substitutes), amount and/or timing of feeds, for example, initiation of feeding, and frequency; * regulation of body temperature; * monitoring (including screening) of neonates, including blood or plasma glucose levels and signs and symptoms of hypoglycaemia. Interventions that required initiation by a medical practitioner were excluded from the review. TYPES OF OUTCOME MEASURES: Outcomes that were of interest included: * occurrence of hypoglycaemia; * re-establishment and maintenance of blood or plasma glucose levels at or above set threshold (as defined by the particular study); * successful breast-feeding; * developmental outcomes. TYPES OF RESEARCH DESIGNS: The review initially focused on randomised controlled trials reported from 1995 to 2004. Insufficient randomised controlled trials were identified and the review was expanded to include additional cohort and cross-sectional studies for possible inclusion in a narrative summary. SEARCH STRATEGY: The major electronic databases, including MEDLINE/PubMed, CINAHL, EMBASE, LILACS, Cochrane Library, etc., were searched using accepted search techniques to identify relevant published and unpublished studies undertaken between 1995 and 2004. Efforts were made to locate any relevant unpublished materials, such as conference papers, research reports and dissertations. Printed journals were hand-searched and reference lists checked for potentially useful research. The year 1995 was selected as the starting point in order to identify any research that had not been included in the World Health Organisation review, which covered literature published up to 1996. The search was not limited to English language studies. ASSESSMENT OF QUALITY: Three primary reviewers conducted the review assisted by a review panel. The review panel was comprised of nine nurses with expertise in neonatal care drawn from senior staff in several metropolitan neonatal units and education programs. Authorship of journal articles was not concealed from the reviewers. Methodological quality of each study that met the inclusion criteria was assessed by two reviewers, using a quality assessment checklist developed for the review. Disagreements between reviewers were resolved through discussion or with the assistance of a third reviewer. DATA EXTRACTION AND ANALYSIS: Two reviewers used a data extraction form to independently extract data relating to the study design, setting and participants; study focus and intervention(s); and measurements and outcomes. As only one relevant randomised controlled trial was found, a meta-analysis could not be conducted nor tables constructed to illustrate comparisons between studies. Instead, the findings were summarised by a narrative identifying any relevant findings that emerged from the data. RESULTS: Seven studies met the inclusion criteria for the objective of this systematic review. The review provided information on the effectiveness of three categories of intervention - type of feeds, timing of feeds and thermoregulation on two of the outcome measures identified in the review protocol - prevention of hypoglycaemia, and re establishment and maintenance of blood or plasma glucose levels above the set threshold (as determined by the particular study). There was no evidence available on which to base conclusions for effectiveness of monitoring or developmental outcomes, and insufficient evidence for breast-feeding success. Given that only a narrative review was possible, the findings of this review should be interpreted with caution. The findings suggest that the incidence of hypoglycaemia in healthy, breast-fed term infants of appropriate size for gestational age is uncommon and routine screening of these infants is not indicated. The method and timing of early feeding has little or no influence on the neonatal blood glucose measurement at 1 h in normal term babies. In healthy, breast-fed term infants the initiation and timing of feeds in the first 6 h of life has no significant influence on plasma glucose levels. The colostrum of primiparous mothers provides sufficient nutrition for the infant in the first 24 h after birth, and supplemental feeds or extra water is unnecessary. Skin-to-skin contact appears to provide an optimal environment for fetal to neonatal adaptation after birth and can help to maintain body temperature and adequate blood glucose levels in healthy term newborn infants, as well as providing an ideal opportunity to establish early bonding behaviours. IMPLICATIONS FOR PRACTICE: The seven studies analysed in this review confirm the World Health Organisation's first three recommendations for prevention and management of asymptomatic hypoglycaemia, namely: 1 Early and exclusive breast-feeding is safe to meet the nutritional needs of healthy term newborns worldwide. 2 Healthy term newborns that are breast-fed on demand need not have their blood glucose routinely checked and need no supplementary foods or fluids. 3 Healthy term newborns do not develop 'symptomatic' hypoglycaemia as a result of simple underfeeding. If an infant develops signs suggesting hypoglycaemia, look for an underlying condition. Detection and treatment of the cause are as important as correction of the blood glucose level. If there are any concerns that the newborn infant might be hypoglycaemic it should be given another feed. Given the importance of thermoregulation, skin-to-skin contact should be promoted and 'kangaroo care' encouraged in the first 24 h after birth. While it is important to main the infant's body temperature care should be taken to ensure that the child does not become overheated. PMID- 21631751 TI - Enhancing the care of the elderly. PMID- 21631749 TI - The JBI model of evidence-based healthcare. AB - Evidence-based healthcare as it is contemporarily conceived is based on the view that clinical decisions should be based on the best available scientific evidence but recognising patient preferences, the context of healthcare and the judgement of the clinician. The ongoing debate on the nature of evidence for practice across all of the health professions is influenced by the experience of clinicians in everyday practice who, in using the evidence, assert that there are diverse sources of research-based and non-research-based evidence and that the process of evidence-based practice should be placed within a broader context that is grounded in practice; recognises different evidentiary bases; and is directed towards improving global health across vasty different practice contexts. We present a developmental framework of evidence-based practice that builds and expands on the work of leaders in the field of evidence-based healthcare; is contextualised; is inclusive of diverse forms of evidence; and incorporates understandings of knowledge transfer and utilisation. The conceptual model attempts to situate healthcare evidence and its role and use within the complexity of practice settings globally. PMID- 21631752 TI - Strategies to reduce medication errors with reference to older adults. AB - Background In Australia, around 59% of the general population uses prescription medication with this number increasing to about 86% in those aged 65 and over and 83% of the population over 85 using two or more medications simultaneously. A recent report suggests that between 2% and 3% of all hospital admissions in Australia may be medication related with older Australians at higher risk because of higher levels of medicine intake and increased likelihood of being admitted to hospital. The most common medication errors encountered in hospitals in Australia are prescription/medication ordering errors, dispensing, administration and medication recording errors. Contributing factors to these errors have largely not been reported in the hospital environment. In the community, inappropriate drugs, prescribing errors, administration errors, and inappropriate dose errors are most common. Objectives To present the best available evidence for strategies to prevent or reduce the incidence of medication errors associated with the prescribing, dispensing and administration of medicines in the older persons in the acute, subacute and residential care settings, with specific attention to persons aged 65 years and over. Search strategy Bibliographic databases PubMed, Embase, Current contents, The Cochrane Library and others were searched from 1986 to present along with existing health technology websites. The reference lists of included studies and reviews were searched for any additional literature. Selection criteria Systematic reviews, randomised controlled trials and other research methods such as non-randomised controlled trials, longitudinal studies, cohort or case-control studies, or descriptive studies that evaluate strategies to identify and manage medication incidents. Those people who are involved in the prescribing, dispensing or administering of medication to the older persons (aged 65 years and older) in the acute, subacute or residential care settings were included. Where these studies were limited, evidence available on the general patient population was used. Data collection and analysis Study design and quality were tabulated and relative risks, odds ratios, mean differences and associated 95% confidence intervals were calculated from individual comparative studies containing count data where possible. All other data were presented in a narrative summary. Results Strategies that have some evidence for reducing medication incidents are: * computerised physician ordering entry systems combined with clinical decision support systems; * individual medication supply systems when compared with other dispensing systems such as ward stock approaches; * use of clinical pharmacists in the inpatient setting; * checking of medication orders by two nurses before dispensing medication; * a Medication Administration Review and Safety committee; and * providing bedside glucose monitors and educating nurses on importance of timely insulin administration. In general, the evidence for the effectiveness of intervention strategies to reduce the incidence of medication errors is weak and high-quality controlled trials are needed in all areas of medication prescription and delivery. PMID- 21631753 TI - Developing clinical leaders in Australian aged care homes. AB - The role of clinical leadership in implementing evidence based practice is increasingly recognised in the health and social care fields. This paper briefly reviews the literature on clinical leadership and evidence-based practice in aged care and describes the established of an aged care clinical fellowship program in Australia. The purpose of this paper is to introduce the reports of four aged care clinical fellows reported elsewhere in this issue of the International Journal of Evidence-Based Healthcare. PMID- 21631754 TI - The prevention and management of constipation in older adults in a residential aged care facility. AB - The need to implement programs for developing leadership and practice improvement skills using an evidence-based practice approach to practice change is becoming more apparent in the health and aged care services. This is no more apparent than in high care residential health and aged care services, where health professionals are increasingly required to provide care for older people with multifocal and complex healthcare needs. This paper describes one of the projects undertaken as part of the Joanna Briggs Institute Commonwealth Department of Health and Ageing Clinical Aged Care Fellowship program from February 2005 to June 2005. This purpose of this particular project was twofold. First it sought to improve the local practice in the prevention and management of constipation and that this practice was performed according to the best available evidence. Second to use the Joanna Briggs Institute Practical Application of Clinical Guidance (PACES) program to implement a process of audit and feedback as a strategy to improve practice. The project was designed to link in with the facility's existing quality improvement program and better practice continence management project. The project was conducted over 6 months and was divided into six stages involving the identification of evidence-based standards of care, an initial audit to determine appropriate sample size, a clinical audit across the facility, planning of the implementation process, implementation of the action plan and re-audit to assess practice change. Overall, the results were extremely positive and demonstrated a real improvement in practice relating to constipation in the project facility. This success, however, needs to be seen in the context of the benefits of having the support of senior management, an existing quality improvement and continence management better practice project, and a culture of clinical review. Although there will always be more work to be done, the success of this project can be viewed in terms of the improvements gained and the long term benefits for the facility and the organisation using the time-efficient audit and feedback strategy. PMID- 21631755 TI - Improving the oral health of older adults with dementia/cognitive impairment living in a residential aged care facility. AB - Background Studies conducted in Residential Aged Care Facilities (RACFs) indicate that high levels of oral diseases and conditions are prevalent in older adults who have dementia. Poor oral health impacts on eating ability, weight, speech, hydration, severity of behavioural problems, appearance and social interactions. This study looked at a group of older adults with dementia in a RACF site in the northern suburbs of Adelaide, South Australia. It is known that in nursing home residents with dementia dental pain and problems are under detected and under-treated. Strategy An audit was conducted to assess the level of compliance of the RACFs oral hygiene care practices with established best practice. The audit questions were based on current best practice as identified from a rigorous international systematic review of the subject. A clinical audit software program (The Joanna Briggs Institute, Practical Application of Clinical Evidence System (JBI PACES)) was used to manage the audit. An audit, feedback, re audit cycle was followed. Stakeholders of the project were identified from which a Project team was formed. The Project team analysed the results of the first audit, conducted a situational analysis and formulated and implemented a strategic plan to target specific criteria for a change management process. Short term and longer-term strategies were identified. Those criteria targeted as achievable in the short term were then re-audited after 6 weeks to determine the effectiveness of the change management process. Findings The criterion Daily cleaning and night-time removal of dentures are documented was re-audited and although there was a slight increase in compliance across the site this increase was not statistically significant. The criterion Resident's dentures are individually marked was re-audited and showed a large increase in compliance across the site, this increase was statistically significant. PMID- 21631756 TI - Compliance with best practice in oral health: implementing evidence in residential aged care. AB - Background The changing oral health status and the increasing complexity of dental needs for older Australians highlights the importance of oral healthcare within the residential aged care setting. The increasing incidence of oral disease and conditions in older adults is related to higher levels of oral functional dependence, physical frailty, medical conditions, polypharmacy, cognitive impairment and increasing dependence on carers. Objectives The Compliance with Best Practice in Oral Health Project was designed to identify issues in relation to oral healthcare and to identify that evidence-based best practice was demonstrated within the organisation's existing policies and procedures. Evidence-based best practice recommendations were based on the systematic evidence review 'oral hygiene care for adults with dementia in residential aged care' developed by the Joanna Briggs Institute. Results Four aged care facilities participated in the project. In the pre-audit, compliance was achieved in one of the six evidence-based criteria. A plan was implemented to improve compliance with three of the remaining five criteria within a 4-week time frame. Post-audit results indicated an improvement in compliance with evidence based best practice. Conclusion The Compliance with Best Practice in Oral Health Project provided an opportunity to review and update existing organisational clinical care policies with evidence-based best practice recommendations and highlighted the importance of developing local clinical care leadership at the residential site. PMID- 21631757 TI - Maintaining oral hydration in older adults living in residential aged care facilities. AB - Background The Clinical Fellowship program was developed to address the need for best practice in aged care and therefore improve outcomes for residents in Residential Aged Care. The project chosen for this program was Oral Hydration with the aim of improving the hydration status of Nursing Home residents and management of those at risk of dehydration. Method The clinical audit tool used was the Joanna Briggs Institute Practical Application of Clinical Evidence System (JBI-PACES). This tool allows measurement of current practice against best practice. The Getting Research into Practice program was then used to address non compliance with best practice. This program requires consultation with key stakeholders to identify limitations to achieving best practice and plan action to address these limitations where possible. Conclusion While 100% compliance with all audit criteria was not achieved, strategies to improve current practice were identified and are in the process of being implemented. The tools developed by The JBI have been beneficial in identifying, measuring and implementing best practice. The process has the potential to contribute to improved resident outcomes and best practice in residential aged care through clinical audit and feedback. PMID- 21631758 TI - Of hedgehogs and foxes: issues in the review and utilisation of research-based evidence. PMID- 21631759 TI - Barriers to, and facilitators of, research utilisation: a survey of Hong Kong registered nurses. AB - Aim Despite increasing efforts to promote the utilisation of research in daily nursing practice it is apparent that there are a number of obstacles to address. This paper reports a study to examine the barriers to, and facilitators of, research utilisation among registered nurses in Hong Kong. Methods A survey design and a random sampling method was used. The final sample consisted of 1487 registered nurses working in private and public health-care sectors in Hong Kong. A bilingual version of the Research Utilisation Questionnaire, comprising a 31 item barriers scale, and an 8-item facilitators scale was used. The instrument was mailed to participants who were asked to return the completed questionnaire by mail. Results The highest ranking barriers to research utilisation reported by respondents were related mainly to organisational factors with regards to inadequate facilities, no authority to change procedures, and time constraints. Hong Kong nurses, however, did not appear to see any problem with regards to items related to characteristics of research, such as conclusions drawn from research being justified, research articles not being published fast enough, and literature reporting conflicting results. This indicates that nurses are aware of research developments in nursing and can critically analyse research reports. With regards to facilitators of research utilisation, respondents agreed that managerial and peer support are the greatest facilitators. Conclusions The results indicate that factors influencing research utilisation are multidimensional and should be taken into account by all involved in the research enterprise: researchers, practitioners, educators, managers and policy-makers. The results of this study provide directions on how to assist nurses in Hong Kong in their efforts to utilise research. PMID- 21631760 TI - Support for overseas qualified nurses in adjusting to Australian nursing practice: a systematic review. AB - Objectives The objective of the review was to summarise the best available evidence supporting overseas nurses' adjustment to Australian nursing practice. The specific review question was: what supportive interventions assist overseas nurses to adjust to Australian nursing practice? Inclusion criteria The review considered qualitative and quantitative papers that addressed adjustment issues of overseas qualified nurses coming and working in Australia. The types of participants were nurses who have received basic nursing education outside Australian, and either nurses who already registered and were working as nurses in Australia or nurses who were undertaking courses required for registration in Australia. Interventions of interest included creating positive work and educational environments that support overseas qualified overseas nurses' adjustment to nursing practice in Australia. Search strategy The search strategy sought to find both published and unpublished papers in English language. The search was performed using the following databases: Medline, CINAHL, ERIC, AUSTRUM, APAIS, Sociological Abstract, ProQuest, Dissertation Abstract. In addition, the reference lists and bibliographies of the articles were also hand searched to identify other studies. Relevant worldwide websites were also searched. Methodological quality Each paper was assessed by two reviewers for methodological quality prior to inclusion in the review using a critical appraisal instrument from Qualitative Assessment and Review Instrument (QARI) software developed by the Joanna Briggs Institute (JBI). Results A total of 12 papers, qualitative, quantitative and textual in nature, were included in the review. Sixty-four papers were identified and 52 papers were excluded as they did not meet the inclusion criteria. There were: three papers utilising qualitative methodology (two phenomenology, one grounded theory), three program evaluation reports, two descriptive studies, and four expert opinions being included. Findings from two qualitative studies utilising a phenomenological approach were extracted and meta-synthesised using JBI-QARI. Two syntheses were derived: (i) overseas nurses find entry into Australian culture very difficult; and (ii) overseas nurses who feel lonely, isolated or 'outsiders' experience difficulty in settling in to nursing in Australia. Findings from other papers were discussed in narrative form. Conclusion The clash of cultures between overseas nurses and the dominant Australian culture should be addressed in a transition program. If strategies to assist overseas nurses to establish informal networks of friends and professional colleagues are in place, the transition to becoming effective practitioners could be greatly enhanced. PMID- 21631761 TI - Effectiveness of standard short-course chemotherapy for treating tuberculosis and the impact of drug resistance on its outcome. AB - Background Treating tuberculosis (TB) with short-course chemotherapy is recommended by the World Health Organization tuberculosis program and is one of the five packages of directly observed treatment short-course (DOTS) strategy. Objectives To investigate the effectiveness of short-course chemotherapy for treating new and retreatment TB patients and to assess the impact of drug resistance on its outcomes. Search strategy and selection criteria Published studies from the electronic databases such as Cochrane Library, DARE, Medline, Embase, Current Contents, CINAHL, Expanded Academic Index, PsycInfo, AustHealth and unpublished studies from Dissertation Abstract International, Index to These, Cochrane Reviewer Handbook were searched between 1993-2002. Studies addressing the treatment of patients with short-course regimen and the association between drug resistance and treatment outcomes were included. Two reviewers independently assessed study quality and extracted data. Results and discussion Eleven cohort studies met the inclusion criteria. Treatment with 6-month daily regimens was effective for new TB patients with success rate (percentage of cure cases and treatment completed cases) of over 80%. Implementation of 100% directly observed treatment was necessary to achieve over 80% treatment success in new cases treated with the intermittent regimen. The intermittent regimens did not work effectively for retreatment cases since success rate was only 68.5% even in the setting using 100% directly observed treatment in Peru. Short-course regimens were effective against drug-sensitive TB with the success rate of 87%; ineffective for patients with multi-drug resistance (rate of treatment failure in new cases ranged 22.3%-35.5% and rate of treatment failure in retreatment cases ranged 16.3-37.1%). Drug resistance had a negative impact on the outcome of short course therapy and multi-drug resistance had a huge negative impact on the effectiveness of chemotherapy for TB. Conclusion The evidence suggests that the World Health Organization-targeted cure rate of 85% in new smear-positive TB cases is not achievable using the intermittent regimen and the target rate should be adjusted. DOTS-plus multi-drug resistant TB program of the world Health Organization should be adopted. However, the evidence is limited and the level of evidence was III-2. PMID- 21631762 TI - A comprehensive systematic review of evidence on the structure, process, characteristics and composition of a nursing team that fosters a healthy work environment. AB - Objectives The overall aim of this comprehensive systematic review was to identify the best available evidence on the effect of team characteristics, processes, structure and composition within the context of collaborative practice among nursing teams that create a healthy work environment. Search strategy The search strategy sought to find both published and unpublished studies and papers written in the English language. An initial limited search of Medline and CINAHL databases was undertaken to identify optimal search terms. A second extensive search using all identified keywords and index terms was then undertaken. Methodological quality Two independent reviewers assessed the methodological quality of retrieved papers using the corresponding checklist from the System for the Unified Management, Assessment and Review of Information (SUMARI) package. Results The papers included in the review included nine experimental or quasi experimental studies, 11 descriptive studies and four qualitative studies. A variety of different team structures such as interdisciplinary teams, primary nursing, team nursing, multidisciplinary models of care delivery and the use of a Partner in Patient Care model were investigated. Team characteristics should include accountability, commitment, enthusiasm and motivation. Social support within a team from a supervisor or colleague increased satisfaction levels of staff. Conclusion The results of the review lead to the development of a number of recommendations for practice that could assist with creating a health work environment. PMID- 21631763 TI - The bankruptcy of evidence- based practice? PMID- 21631764 TI - Implementation of oral health recommendations into two residential aged care facilities in a regional Australian city. AB - Background Residents of aged care facilities usually have a large number of oral health problems. Residents who suffer from dementia are at particular risk. A systematic review of the best available evidence with regard to maintaining the oral health of older people with dementia in residential aged care facilities provided a number of recommendations. Objectives The aim of the implementation project was to introduce evidence-based oral hygiene practices for patients with dementia in two publicly funded residential aged care facilities and monitor for changes in nursing awareness, knowledge, documentation and practice to improve patient outcomes and ensure appropriate accreditation standards were met. An additional aim was to identify barriers and strategies to overcome barriers to implementation of evidence-based recommendations. Methods Two facilities, a 40 bed facility and a 71-bed facility in the health service district of the regional Australian city of Toowoomba, provided the setting. A quality improvement approach was taken, using a number of strategies from the National Health and Medical Research Council guidelines for implementation studies. The implementation involved a number of stages, including project development, interactive oral health education, oral audits of residents, changes to oral hygiene practice via care plans and critical reflection. Results The multidisciplinary approach to improving oral healthcare appeared to improve knowledge and awareness and move oral health practices in facilities closer to best practice. Specialised training in oral health was provided to a Clinical Nurse Consultant. Regular oral audits were introduced and facility staff were trained in the use of the oral audit tool. Care plans at one facility were of better quality and more comprehensive than before the intervention. Comments made during critical reflection suggested improvements in the oral health of residents, increased use of oral swabs and saliva substitutes, improved care of dentures and mention of the use of mouth props in resident care plans. There was also some evidence that changes brought about by the implementation are sustainable. Conclusion The majority of recommendations provided in the systematic review of oral healthcare for dementia patients were applicable to the applied context. The importance of day-to-day leaders was highlighted by the apparently varied outcomes across target facilities. The quality improvement approach would appear to have considerable advantages when applied to improving practice in residential aged care. PMID- 21631765 TI - Deconstructing the evidence-based discourse in health sciences: truth, power and fascism. AB - Background Drawing on the work of the late French philosophers Deleuze and Guattari, the objective of this paper is to demonstrate that the evidence-based movement in the health sciences is outrageously exclusionary and dangerously normative with regards to scientific knowledge. As such, we assert that the evidence-based movement in health sciences constitutes a good example of microfascism at play in the contemporary scientific arena. Objective The philosophical work of Deleuze and Guattari proves to be useful in showing how health sciences are colonised (territorialised) by an all-encompassing scientific research paradigm - that of post-positivism - but also and foremost in showing the process by which a dominant ideology comes to exclude alternative forms of knowledge, therefore acting as a fascist structure. Conclusion The Cochrane Group, among others, has created a hierarchy that has been endorsed by many academic institutions, and that serves to (re)produce the exclusion of certain forms of research. Because 'regimes of truth' such as the evidence-based movement currently enjoy a privileged status, scholars have not only a scientific duty, but also an ethical obligation to deconstruct these regimes of power. PMID- 21631766 TI - Never mind the evidence, feel the width: a response to Holmes, Murray, Perron and Rail. PMID- 21631767 TI - Entertaining fascism? PMID- 21631768 TI - A comprehensive systematic review of evidence on the effectiveness and appropriateness of undergraduate nursing curricula. AB - Objectives The objective of this review was to appraise and synthesise the best available evidence on the effectiveness and appropriateness of undergraduate nursing curricula. Inclusion criteria This review considered research papers that addressed the effectiveness and appropriateness of undergraduate nursing curricula. Studies of higher evidence levels were given priority over lower evidence studies. Participants of interest were undergraduate nursing students, nursing staff and healthcare consumers. Nursing staff outcomes, consumer outcomes and system outcomes (e.g. competency, satisfaction, critical thinking skills, healthcare consumer rights and cost-effectiveness) that impact on the evaluation of undergraduate nursing curricula were considered in the review. Search strategy The search strategy sought to find both published and unpublished studies and reports limited to the English language. An initial limited search of MEDLINE and CINAHL was undertaken, followed by an analysis of the text contained in the title and abstract, and of the index terms used to describe the article. A second extensive search was then undertaken using all identified key words and index terms. Finally, the reference list of all identified reports and articles was searched for additional studies. Methodological quality Each paper was assessed by two independent reviewers for methodological quality before inclusion in the review using an appropriate critical appraisal instrument from the Unified Management, Assessment and Review of Information (SUMARI) package. Results A total of 16 papers, experimental and textual in nature, were included in the review. The majority of papers was descriptive and examined the relationships between nursing curricula and specific learning outcomes such as critical thinking skills. Because of the diverse nature of these papers, meta-analysis of the results was not possible and this section of the review is presented in narrative form. In this review, four undergraduate nursing curriculum models were identified: integrated curriculum, subject-centred curriculum, problem-based learning, and an integrated critical thinking model. It was possible to examine the effectiveness of an integrated curriculum model and a subject-centred curriculum model; however, the other two models could not be compared because of a lack of evidence. Conclusion The evidence regarding the effectiveness and appropriateness of undergraduate nursing curricula is notably weak because of the paucity of high-quality comparative studies and meaningful outcome measures of available studies. Therefore, no strong conclusion can be made regarding the effectiveness and appropriateness of undergraduate nursing curricula. PMID- 21631769 TI - The feasibility and appropriateness of introducing nursing curricula from developed countries into developing countries: a comprehensive systematic review. AB - Objectives The objective of this review was to appraise and synthesise the best available evidence on the feasibility and appropriateness of introducing nursing curricula from developed countries into developing countries. Inclusion criteria This review considered quantitative and qualitative research papers that addressed the feasibility and appropriateness of introducing developed countries' nursing curricula into developing countries. Papers of the highest level of evidence rating were given priority. Participants of interest were all levels of nursing staff, nursing students, healthcare consumers and healthcare administrators. Outcomes of interest that are relevant to the evaluation of undergraduate nursing curricula were considered in the review including cost effectiveness, cultural relevancy, adaptability, consumer satisfaction and student satisfaction. Search strategy The search strategy sought to find both published and unpublished studies and papers, limited to the English language. An initial limited search of MEDLINE and CINAHL was undertaken followed by an analysis of the text words contained in the title and abstract, and of the index terms used to describe the article. A second extensive search was then undertaken using all identified key words and index terms. Finally, the reference list of all identified reports and articles was searched, the contents pages of a few relevant journals were hand searched and experts in the field were contacted to find any relevant studies missed from the first two searches. Methodological quality Each paper was assessed by two independent reviewers for methodological quality before inclusion in the review using an appropriate critical appraisal instrument from the System for the Unified Management, Assessment and Review of Information (SUMARI) package. Results A total of four papers, including one descriptive study and three textual papers, were included in the review. Because of the diverse nature of these papers, meta-synthesis of the results was not possible. For this reason, this section of the review is presented in narrative form. In this review, a descriptive study and a textual opinion paper examined the cultural relevancy of borrowed curriculum models, and the global influence of American nursing. Another two opinion papers evaluated the adaptability of another country curriculum models in their countries. Conclusion The evidence regarding the feasibility and appropriateness of introducing developed countries' nursing curricula into developing countries is weak because of the paucity of high-quality studies. However, some lower-level evidence suggesting that direct transfer of the curriculum model from one country to another is not appropriate without first assessing the cultural context of both countries. Second, the approach of considering international, regional and local experiences more feasible and presumably a more effective strategy for adapting of a country's curriculum into a culturally or economically different country. PMID- 21631771 TI - Evidence-based healthcare: the role of our Journal. PMID- 21631770 TI - A systematic review of evidence on the professional practice of the nurse and developing and sustaining a healthy work environment in healthcare. AB - Objectives The overall aim of this systematic review was to identify the best available evidence on the relationship between the knowledge, competencies and behaviours of nurses exhibiting professional practice in their workplace; and the development of a healthy work environment. Search strategy The search strategy sought to find both published and unpublished studies written in the English language. An initial limited search of MEDLINE and CINAHL databases was undertaken to identify optimal search terms. A second extensive search using all identified key words and index terms was then undertaken. Methodological quality Two independent reviewers assessed the methodological quality of retrieved papers using the corresponding checklist from the System for the Unified Management, Assessment and Review of Information (SUMARI) package. Results Of the 4238 papers found in the search: 219 were selected for full paper retrieval; 19 of these papers were unable to be located; 200 full papers were assessed for methodological quality; 181 studies were excluded; and 19 were included in the review. Of these 19, four quantitative studies evaluated a professional nursing practice model and its impact on a variety of outcomes; five descriptive studies examined elements of nurses' professional practice and the impact these elements had on specified outcomes; and 10 qualitative papers examined varying behaviours, competencies and knowledge levels of nurses. Various methods were used, such as focus groups and open-ended interviews. Overall, the evidence suggests that professional practice has a positive impact on the work environment in terms of nurses' role satisfaction and patient outcomes. The evidence is, however, equivocal in many areas and the impact of the professional practice of the nurse requires further investigation. Conclusion The results of the review suggest a number of recommendations for practice and research on creating a healthy work environment. PMID- 21631772 TI - Lack of evidence of the effectiveness of primary brachial plexus surgery for infants (under the age of two years) diagnosed with obstetric brachial plexus palsy. AB - Background Obstetric brachial plexus palsy, which occurs in 1-3 per 1000 live births, results from traction and/or compression of the brachial plexus in utero, during descent through the birth canal or during delivery. This results in a spectrum of injuries that range in extent of damage and severity and can lead to a lifelong impairment and functional difficulties associated with the use of the affected upper limb. Most infants diagnosed with obstetric brachial plexus palsy receive treatment, such as surgery to the brachial plexus, physiotherapy or occupational therapy, within the first months of life. However, there is controversy regarding the most effective form of management. This review follows on from our previous systematic review which investigated the effectiveness of primary conservative management in infants with obstetric brachial plexus palsy. This systematic review focuses on the effects of primary surgery. Objectives The objective of this review was to systematically assess and collate all available evidence on effectiveness of primary brachial plexus surgery for infants with obstetric brachial plexus palsy. Search strategy A systematic literature search was performed using 13 databases: TRIP, MEDLINE, CINAHL, Web of Science, Proquest 5000, Evidence Based Medicine Reviews, Expanded Academic ASAP, Meditext, Science Direct, the Physiotherapy Evidence Database, Proquest Digital Dissertations, Open Archives Initiative Search Engine, the Australian Digital Thesis program. Those studies that were reported in English and published between July 1992 to June 2004 were included in this review. Selection criteria Quantitative studies that investigated the effectiveness of primary brachial plexus surgery for infants with obstetric brachial plexus palsy were eligible for inclusion into this review. This excluded studies where infants were solely managed conservatively or with pharmacological agents, or underwent surgery for the management of secondary deformities. Data collection and analysis Two independent reviewers assessed the eligibility of each study for inclusion into the review, the study design used and its methodological quality. Where any disagreement occurred, consensus was reached by discussion. Studies were also assessed for clinical homogeneity by considering populations, interventions and outcomes. Where heterogeneity was present, synthesis was undertaken in a narrative format. Results Twenty-one studies were included in the review. Most were ranked low on the hierarchy of evidence (no randomised controlled trials were found), and most had only fair methodological quality. Surgical intervention was variable, as were the eligibility criteria for surgery, the timing of surgery and the outcome instruments used to evaluate the effect of surgery. Therefore, it is difficult to draw conclusions regarding the effectiveness of primary brachial plexus surgery for infants with obstetric brachial plexus palsy. Conclusions Although there is a wealth of information regarding the outcome following primary brachial plexus surgery it was not possible to determine whether this treatment is effective in increasing functional recovery in infants with obstetric brachial plexus palsy. Further research is required to develop standardised surgical criteria, and standardised outcome measures should be used at specific points in time during the recovery process to facilitate comparison between studies. Moreover, comparison groups are required to determine the relative effectiveness of surgery compared with other forms of management. PMID- 21631773 TI - Factors associated with constructive staff-family relationships in the care of older adults in the institutional setting. AB - Background Modern healthcare philosophy espouses the virtues of holistic care and acknowledges that family involvement is appropriate and something to be encouraged due to the role it plays in physical and emotional healing. In the aged care sector, the involvement of families is a strong guarantee of a resident's well-being. The important role family plays in the support and care of the older adult in the residential aged care environment has been enshrined in the Australian Commonwealth Charter of Residents' Rights and Responsibilities and the Aged Care Standards of Practice. Despite wide acknowledgement of the importance of family involvement in the healthcare of the older adult, many barriers to the implementation of participatory family care have been identified in past research. For older adults in the healthcare environment to benefit from the involvement of their family members, healthcare professionals need an understanding of the issues surrounding family presence in the healthcare environment and the strategies to best support it. Objectives The objectives of the systematic review were to present the best available evidence on the strategies, practices and organisational characteristics that promote constructive staff-family relationships in the care of older adults in the healthcare setting. Specifically this review sought to investigate how staff and family members perceive their relationships with each other; staff characteristics that promote constructive relationships with the family; and interventions that support staff-family relationships. Search strategy A literature search was performed using the following databases for the years 1990 2005: Ageline, APAIS Health, Australian Family and Society Abstracts (FAMILY), CINAHL, Cochrane Library, Dare, Dissertation Abstracts, Embase, MEDLINE, PsycINFO and Social Science Index. Personal communication from expert panel members was also used to identify studies for inclusion. A second search stage was conducted through review of reference lists of studies retrieved during the first search stage. The search was limited to published and unpublished material in English language. Selection criteria The review was limited to studies involving residents and patients within acute, subacute, rehabilitation and residential settings, aged over 65 years, their family and healthcare staff. Papers addressing family members and healthcare staff perceptions of their relationships with each other were considered for this review. Studies in this review also included those relating to interventions to promote constructive staff-family relationships including organisational strategies, staff-family meetings, case conferencing, environmental approaches, etc. The review considered both quantitative and qualitative research and opinion papers for inclusion. Data collection and analysis All retrieved papers were critically appraised for eligibility for inclusion and methodological quality independently by two reviewers, and the same reviewers collected details of eligible research. Appraisal forms and data extraction forms designed by the Joanna Briggs Institute as part of the QARI and NOTARI systematic review software packages were used for this review. Findings Family members' perceptions of their relationships with staff showed that a strong focus was placed on opportunities for the family to be involved in the patient's care. Staff members also expressed a theoretical support for the collaborative process, however, this belief often did not translate to the staff members' clinical practice. In the studies included in the review staff were frequently found to rely on traditional medical models of care in their clinical practice and maintaining control over the environment, rather than fully collaborating with families. Four factors were found to be essential to interventions designed to support a collaborative partnership between family members and healthcare staff: communication, information, education and administrative support. Based on the evidence analysed in this systematic review, staff and family education on relationship development, power and control issues, communication skills and negotiating techniques is essential to promoting constructive staff-family relationships. Managerial support, such as addressing workloads and staffing issues; introducing care models focused on collaboration with families; and providing practical support for staff education, is essential to gaining sustained benefits from interventions designed to promote constructive family-staff relationships. PMID- 21631774 TI - Systematic review of evidence on the impact of nursing workload and staffing on establishing healthy work environments. AB - Background This systematic review set out to examine the impact, if any, of nursing workload and staffing on creating and maintaining healthy work environments. For the purposes of this review, the term 'healthy work environment' was defined as '. . . a practice setting that maximizes the health and well-being of nurses, quality patient outcomes and organizational performance'. This definition identifies nurse, patient and organisational outcomes as indicators of the establishment and maintenance of a healthy work environment. Objectives The review sought to determine the impact of: * Patient characteristics, nurse characteristics, system characteristics and system processes on workload, scheduling and concepts of productivity and utilisation * Workload, scheduling and concepts of productivity and utilisation on the quality of outcomes for clients, nurses and the system/organisation Search strategy The search strategy sought to find both published and unpublished studies and papers written in the English language. A three-step search strategy approach was used. An initial limited search of MEDLINE and CINAHL databases was undertaken to identify optimal search terms followed by an analysis of the text words contained in the title and abstract, and of the index terms used to describe the article. A second extensive search using all identified keywords and index terms was then undertaken. The third step consisted of a search of the reference lists of all identified reports and articles for additional studies. Selection criteria Types of studies: This review considered research papers that addressed the appropriateness and effectiveness of workload and staffing concepts in fostering a healthy work environment in healthcare. The types of papers to be considered included: meta-analysis, randomised controlled trials, quasi-randomised controlled trials, cohort studies, case-control studies, descriptive studies and correlational studies. TYPES OF PARTICIPANTS: The review considered all participants involved or affected by workload and staffing concepts within the nursing workforce in a healthcare environment, including staff and patients. System and policy issues were also considered. Types of interventions: All workload and staffing strategies that impact on the work environment, patient and nurse outcomes were considered in this review. Types of outcome measures: Outcomes of interest were categorised into four groups: nursing staff outcomes, patient outcomes, organisational outcomes and system outcomes. Data collection and analysis Following assessment of methodological quality, data were extracted using data extraction tools based on the work of the Cochrane Collaboration and the Centre for Reviews and Dissemination. Statistical pooling was not possible and findings were presented in narrative form. Results Of the 2162 papers identified in the search, 171 were selected for full paper retrieval and assessed independently by two reviewers for methodological quality. A total of 40 papers were included in the review: one systematic review; one cohort study; and 38 correlational descriptive studies. Results were summarised in narrative form. The evidence suggests strong correlations between patient characteristics and work environments; and workload and staffing and the quality of outcomes for clients, nurses and the system/organisation. This gave rise to a number of recommendations for practice and for further research, such as: * A greater proportion of regulated staffing (i.e. registered nurses, enrolled nurses, practical or vocational nurses) is associated with improved outcomes related to the Functional Independence Measure score, the Short Form Health Survey (SF-36) vitality score, patient satisfaction with nursing care, patient adverse events (including atelectasis, decubitus ulcers, falls, pneumonia, postsurgical and treatment infection and urinary tract infections) * An increase in the number of registered nurse hours available is associated with improved patient outcomes in relation to falls, pneumonia, pressure ulcers, urinary tract infection, length of stay and postoperative infection rates. PMID- 21631775 TI - Scientists, postmodernists or fascists? AB - The somewhat frenzied reaction to publication of a provocative, discursive paper titled 'Deconstructing the evidence-based discourse in health sciences: truth, power and fascism' by Holmes et al. in the International Journal of Evidence Based Healthcare is both surprising and worrying. The paper is essentially a postmodernist critique of evidence-based healthcare. In the same issue of the journal in which the paper was published both the guest editorial and a response to the paper refute its claims. However, media coverage on the paper gave rise to numerous defensive responses that attacked the paper through claiming it represents 'bad science' or by disparaging the International Journal of Evidence Based Healthcare, its Editor, its peer review processes or the organisation linked to the journal, the Joanna Briggs Institute. It is clear that those who mounted these attacks had no knowledge of the journal (or of the editorial and response refuting the claims made in the paper, published in the same issue) or its parent organisation; and none of them attempted to critique the paper in a scholarly fashion. This paper sets out to construct a scholarly argument to refute these claims and to consider why it is that those who support evidence based healthcare and/or science chose to disparage a journal and an organisation that promotes and facilitates evidence-based approaches to healthcare - and the value of the Cochrane Collaboration - rather than developing a rigorous critique of the argument developed in the Holmes et al. paper. Although this response appears to be an attempt to silence dissenting views (and may, to some, suggest that the reference to microfascism in the paper in question may, indeed, have some validity) we conclude that the postmodernist critique of evidence-based healthcare embodied in the paper sets out criticisms that, though widespread in healthcare, can be challenged in a considered, scholarly way. The ill-informed, reactionary responses to it by the defenders of science make little contribution to the ongoing development of evidence to improve global health. PMID- 21631776 TI - How to grab a headline: lots of smoke and little meat. (Response to Holmes et al.). PMID- 21631777 TI - Towards an understanding of the politics of 'evidence': a response to Dr Jefferson, MD. PMID- 21631778 TI - Some comments on the problems and merits of the evidence-based practice. PMID- 21631780 TI - Best available evidence. PMID- 21631781 TI - Systematic review of interventions in the management of overweight and obese children which include a dietary component. AB - Background The prevalence of overweight and obesity in children and adolescents is increasing at an alarming rate around the world and prevention has become a key public health objective. Treatment and management of those already overweight and obese must be aligned with the best available evidence on effectiveness, if the risk of obesity-related morbidity and mortality is yet be reduced. Diet plays a pivotal role in successful treatment of obesity but to date, there is limited evidence on which to base practice. Objectives To identify and present the best available evidence on the optimal dietetic treatment and management of children and adolescent who are overweight or obese. Search strategy Published English language literature was searched using the electronic databases CINAHL, MEDLINE, PRE-MEDLINE, DARE, COCHRANE, EMBASE, AUSTROM, Current Concepts and Dissertation Abstracts. The databases were limited to English Language from 1975 until 2003. Government reports from the UK, USA and Australian were also searched and a hand search performed for the Journal of the Dietitians Association of Australia, International Journal of Obesity and the Journal of Human Nutrition and Dietetics and the bibliographies of retrieved articles. Selection criteria (i) Interventions that evaluated the effectiveness of nutrition or dietary interventions to treat or manage overweight and obesity; (ii) Children aged less than 18 years; and (iii) Participants were defined as overweight or obese by relative weight or a measure of body weight status, studies that reported body weight per se were excluded. Data collection and analysis An experienced professional librarian searched the databases, and two trained research assistants independently identified studies for retrieval and assessed each article for inclusion. The included studies were critically appraised for methodological quality by two people independently. Data were extracted from the appropriate articles and when a discrepancy arose, a third party would arbitrate. Main results There were 116 articles that met the inclusion criteria. While 49 articles described randomised controlled trials, they arose from 37 separate studies. There were 67 non-randomised trials. Meta-analyses were performed on eight studies that included both a dietary intervention component and an adequate control group and on four studies that had follow-up data. There was a high degree of heterogeneity between studies and this made comparisons between studies problematic. Interventions that include diet therapy generally result in significant weight loss, at least in the short term. Many studies were poorly designed and had no or only minimal follow up. The details of the dietary intervention were often inadequately described and dietary outcomes rarely reported, making repetition of the studies difficult. Reviewers' conclusions There is an urgent need for high quality studies investigating the optimal dietary approach to management of paediatric overweight and obesity. These studies require adequate follow up to ascertain if weight loss can be sustained in the long term. Details of the dietary prescription, adherence to the dietary intervention and diet-specific outcomes need to be reported in order to inform best practice. PMID- 21631782 TI - Systematic review on embracing cultural diversity for developing and sustaining a healthy work environment in healthcare. AB - Objectives The objective of this review was to evaluate evidence on the structures and processes that support development of effective culturally competent practices and a healthy work environment. Culturally competent practices are a congruent set of workforce behaviours, management practices and institutional policies within a practice setting resulting in an organisational environment that is inclusive of cultural and other forms of diversity. Inclusion criteria This review included quantitative and qualitative evidence, with a particular emphasis on identifying systematic reviews and randomised controlled trials. For quantitative evidence, other controlled, and descriptive designs were also included. For qualitative evidence, all methodologies were considered. Participants were staff, patients, and systems or policies that were involved or affected by concepts of cultural competence in the nursing workforce in a healthcare environment. Types of interventions included any strategy that had a cultural competence component, which influenced the work environment, and/or patient and nursing staff in the environment. The types of outcomes of interest to this review included nursing staff outcomes, patient outcomes, organisational outcomes and systems level outcomes. Search strategy The search sought both published and unpublished literature written in the English language. A comprehensive three-step search strategy was used, first to identify appropriate key words, second to combine all optimal key words into a comprehensive search strategy for each database and finally to review the reference lists of all included reviews and research reports. The databases searched were CINAHL, Medline, Current Contents, the Database of Abstracts of Reviews of Effectiveness, The Cochrane Library, PsycINFO, Embase, Sociological Abstracts, Econ lit, ABI/Inform, ERIC and PubMed. The search for unpublished literature used Dissertation Abstracts International. Methodological quality Methodological quality was independently established by two reviewers, using standardised techniques from the Joanna Briggs Institute (JBI) System for the Unified Management, Assessment and Review of Information (SUMARI) package. Discussion with a third reviewer was initiated where a low level of agreement was identified for a particular paper. Following inclusion, data extraction was conducted using standardised data extraction tools from the JBI SUMARI suite for quantitative and qualitative research. Data synthesis was performed using the JBI Qualitative Assessment and Review Instrument and JBI Narrative, Opinion and Text Assessment and Review Instrument software to aggregate findings by identifying commonalities across texts. Quantitative data were presented in narrative summary, as statistical pooling was not appropriate with the included studies. Results Of the 659 identified papers, 45 were selected for full paper retrieval, and 19 were considered to meet the inclusion criteria for this review. The results identified a number of processes that would contribute to the development of a culturally competent workforce. Appropriate and competent linguistic services, and intercultural staff training and education, were identified as key findings in this review. Conclusions The review recommends that health provider agencies establish links with organisations that can address needs of culturally diverse groups of patients, include cultural competence in decision support systems and staff education as well as embed them in patient brochures and educational materials. The review also concluded that staff in-service programs consider the skills needed to foster a culturally competent workforce, and recruitment strategies that also explicitly address this need. PMID- 21631783 TI - Evidence to practice: pre-post-implementation study of a patient/provider resource for self-management with heart failure. AB - Background Evidence-based recommendations for heart failure self-management are contained in quality clinical practice guidelines. To implement these in practice requires additional translation. Partners in Care for Congestive Heart Failure (PCCHF) is a set of resource materials developed to encourage heart failure patients and their families to assume greater responsibility and to participate in daily decision-making related to their illness experience by enhancing their self-assessment and self-management skills. The study objectives were to evaluate its use, acceptability and relevance of this approach by heart failure patients, nurses and policy-makers. Methods A pre-post study was conducted across 10 rural urban, acute and community care sites within three Canadian provinces and one US state. Patients' health-related quality of life was assessed with Minnesota Living with Heart Failure Questionnaire and Medical Outcomes Short Form before and 6 weeks after using the PCCHF program. Nurses completed a survey and participated in focus groups. Policy-makers were interviewed before and post implementation. Results Baseline and 6-week measures were completed by 239 patients. Health-related quality of life measures revealed statistically significant improvement after 6 weeks. Thirty-three nurses and 19 policy-makers participated in interviews post implementation. Most patients, nurses and policy makers found the resource acceptable and relevant to support information needs. Conclusion The PCCHF program positively benefited both patients and clinical staff. The evidence-based teaching materials were considered a useful resource for self-management with heart failure. Time constraints and high staff turnover underline the need for resources like PCCHF to assist in patient-oriented heart failure self-management. Copyright for PCCHF has been transferred to the Canadian Heart and Stroke Foundation for widespread dissemination. PMID- 21631784 TI - Reducing physical restraint use in residential aged care: implementation of an evidence-based approach to improve practice. AB - Background This paper describes the approach taken by three high care residential facilities in reducing physical restraint use and improving practice when physical restraint is used as a care strategy. At the time of the project these three facilities provided care for 174 residents, with over half of the residents having a diagnosis of dementia. Method A process of audit and feedback was used to identify initial compliance with best practice and facilitate identification of barriers to changing and improving clinical care. Results Gaps were found in the areas of organisational documentation (including policy, procedures, forms and information for residents and their families) and staff education on the topic of physical restraint. Once these gaps were known, the task was to develop strategies to address the needs of three stakeholder groups: individual clinicians, residents and their families, and the organisation as a whole. Conclusion The paper shows the advantages of auditing compliance with evidence-based criteria as a measure of improvements in clinical care, and describes the changes implemented as a means of improving practice and reducing physical restraint use. The results show that the process has been effective in achieving results in the short term, with large increases in the area of staff education, and improvements in the assessment of people identified as possibly requiring restraint. Future plans for sustaining and consolidating change and moving forward with improvements are discussed. The results so far have been encouraging and the organisation is committed to continue to improve compliance with best practice with the vision of eventually achieving restraint-free care environments. PMID- 21631785 TI - TLC - Truth, Lies and Consequences. PMID- 21631786 TI - Minimising undernutrition in the older inpatient. AB - Background Malnutrition among elderly hospitalised patients is widespread and has been shown to lead to adverse health outcomes. The effectiveness of interventions to minimise undernutrition in elderly inpatients is not well documented. Objectives To identify the best available practices, in the hospital setting, that minimise undernutrition or the risk of undernutrition, in the acute care patient especially for the older patient. The review will assesses the effectiveness of a range of interventions designed to promote adequate nutritional intake in the acute care setting, with the aim of determining what practices minimise malnutrition in the elderly inpatients. Search strategy English language articles from 1980 onwards were sought using Medline, Premedline, Cinahl, Austrom-Australasian Medical Index and AustHealth, Embase and Science Citations Index. Selection criteria For inclusion the study had to include an intervention aiming to minimise undernutrition in hospitalised elderly patients aged 65 years or older. All study designs were included. Data collection and analysis Two independent reviewers assessed the eligibility of each study for inclusion into the review, critically appraised the study quality and extracted data using standardised tools. For each outcome measure results were tabulated by intervention type and discussed in a narrative summary. Results from randomised controlled trials were pooled in meta-analyses where appropriate. Main results Twenty-nine studies met the inclusion criteria, with a total of 4021 participants. The focus of 15 interventions was the supplying of oral supplements to the participants, six focused on enteral nutrition therapy, four interventions made changes to the foods provided as part of the hospital diet, one included the services of an additional staff member and three incorporated the implementation of evidence-based guidelines. Ten meta-analyses were conducted from which the main findings were: significant improvements in weight status and arm muscle circumferences with an oral supplement intervention, P < 0.05. Reviewers' conclusions The findings of the review support the use of oral supplements to minimise undernutrition in elderly inpatients. The results also emphasise the need for more high-quality research using appropriate outcome measures in the area of minimisation of undernutrition, particularly interventions that make alterations to the hospital diet and address support for feeding patients at the ward level. PMID- 21631787 TI - Effectiveness of arthroscopic versus open surgical stabilisation for the management of traumatic anterior glenohumeral instability. AB - Background Anterior instability is a frequent complication following a traumatic glenohumeral dislocation. Frequently the underlying pathology associated with recurrent instability is a Bankart lesion. Surgical correction of Bankart lesions and other associated pathology is the key to successful treatment. Open surgical glenohumeral stabilisation has been advocated as the gold standard because of consistently low postoperative recurrent instability rates. However, arthroscopic glenohumeral stabilisation could challenge open surgical repair as the gold standard treatment for traumatic anterior glenohumeral instability. Objectives Primary evidence that compared the effectiveness of arthroscopic versus open surgical glenohumeral stabilisation was systematically collated regarding best practice management for adults with traumatic anterior glenohumeral instability. Search strategy A systematic search was performed using 14 databases: MEDLINE, Cumulative Index of Nursing and Allied Health (CINAHL), Allied and Complementary Medicine Database (AMED), ISI Web of Science, Expanded Academic ASAP, Proquest Medical Library, Evidence Based Medicine Reviews, Physiotherapy Evidence Database, TRIP Database, PubMed, ISI Current Contents Connect, Proquest Digital Dissertations, Open Archives Initiative Search Engine, Australian Digital Thesis Program. Studies published between January 1984 and December 2004 were included in this review. No language restrictions were applied. Selection criteria Eligible studies were those that compared the effectiveness of arthroscopic versus open surgical stabilisation for the management of traumatic anterior glenohumeral instability, which had more than 2 years of follow up and used recurrent instability and a functional shoulder questionnaire as primary outcomes. Studies that used non-anatomical open repair techniques, patient groups that were specifically 40 years or older, or had multidirectional instability or other concomitant shoulder pathology were excluded. Data collection and analysis Two independent reviewers assessed the eligibility of each study for inclusion into the review, the study design used and its methodological quality. Where any disagreement occurred, consensus was reached by discussion with an independent researcher. Studies were assessed for homogeneity by considering populations, interventions and outcomes. Where heterogeneity was present, synthesis was undertaken in a narrative format; otherwise a meta-analysis was conducted. Results Eleven studies were included in the review. Two were randomised controlled trials. Evidence comparing arthroscopic and open surgical glenohumeral stabilisation was of poor to fair methodological quality. Hence, the results of primary studies should be interpreted with caution. Observed clinical heterogeneity in populations and outcomes was highlighted and should be considered when interpreting the meta-analysis. Authors also used variable definitions of recurrent instability and a variety of outcome measures, which made it difficult to synthesise results. When comparable data were pooled, there were no significant differences (P > 0.05) between the arthroscopic and open groups with respect to recurrent instability rates, Rowe score, glenohumeral external rotation range and complication rates. Conclusions Statistically, it appears that both surgical techniques are equally effective in managing traumatic anterior glenohumeral instability. In light of the methodological quality of the included studies, it is not possible to validate arthoscopic stabilisation to match open surgical stabilisation as the gold standard treatment. Further research using multicentred randomised controlled trials with sufficient power and instability-specific questionnaires with sound psychometric properties is recommended to build on current evidence. The choice of treatment should be based on multiple factors between the clinician and the patient. PMID- 21631788 TI - Comprehensive systematic review of evidence on developing and sustaining nursing leadership that fosters a healthy work environment in healthcare. AB - Objectives The objective of this review was to appraise and synthesise the best available evidence on the feasibility, meaningfulness and effectiveness of nursing leadership attributes that contribute to the development and sustainability of nursing leadership to foster a healthy work environment. Inclusion criteria This review considered quantitative and qualitative research papers that addressed the feasibility, meaningfulness and effectiveness of developing and sustaining nursing leadership to foster a healthy work environment in healthcare. Papers of the highest level of evidence ratings were given priority. Participants of interest were leaders and those who were affected by leadership, specifically staff and patients. Interventions of interest including positive leadership attributes, as well as system and policy constructs, that impact on the development and sustainability of nursing leadership within the healthcare environment were considered in the review. Search strategy The search strategy sought to find both published and unpublished studies and papers, limited to the English language. An initial limited search of MEDLINE and CINAHL was undertaken followed by an analysis of the text words contained in the title and abstract, and of the index terms used to describe the paper. A second extensive search was then undertaken using all identified key words and index terms. Methodological quality Each paper was assessed by two independent reviewers for methodological quality prior to inclusion in the review using an appropriate critical appraisal instrument from the System for the Unified Management, Assessment and Review of Information (SUMARI) package. Results A total of 48 papers, experimental, qualitative and textual in nature, were included in the review. The majority of papers were descriptive and examined the relationships between leadership styles and characteristics and particular outcomes, such as satisfaction. Because of the diverse nature of these papers meta-analysis of the results was not possible. For this reason, this section of the review was presented in narrative form. The qualitative and textual papers were analysed using The Joanna Briggs Institute-Qualitative Assessment and Review Instrument and The Joanna Briggs Institute-Narrative, Opinion and Text Assessment and Review Instrument. The process of meta-synthesis embodied in these programs involves the aggregation or synthesis of findings or conclusions. Eight syntheses were derived with key themes related to collaboration, education, emotional intelligence, organisational climate, professional development, positive behaviours and qualities and the need for a supportive environment. Conclusion A combination of leadership styles and characteristics was found to contribute to the development and sustainability of a healthy work environment. The current work conducted in this area provides a solid foundation for future directions in research. PMID- 21631789 TI - Evidence-based decision-making: the case for diabetes care. AB - Clinical decisions in diabetes care are complex, because they cover a wide range of unrelated issues which are affected by varied contexts of healthcare providers from multiple disciplines. The purpose of this paper is to reflect on the extent to which evidence is used to underpin clinical decisions within the area of diabetes care. In order to do this, it is necessary to capture information on the application of principles of evidence-based practice in diabetes care. Publications debating the nature of evidence-based practice and diabetes care were identified through searching the MEDLINE and CINAHL databases and located through the search engine Google. Additional publications were identified from references cited in relevant papers. These publications viewed evidence-based decisions as a balanced integration of different aspects of the clinical situation such as patient preference and clinician experience, in addition to the best available evidence. Simultaneously, each aspect of clinical care needs different forms of evidence. This is the argument for evidence-based diabetes care, where heterogeneous variables interact in different milieus. In conclusion, evidence-based diabetes care is a loose structure and set of ideas which needs to be adjusted according to each specific clinical situation. PMID- 21631790 TI - Politics of evidence: conflict and health in Iraq. PMID- 21631791 TI - Critique is crucial. PMID- 21631792 TI - Effectiveness of general practice nurse interventions in cardiac risk factor reduction among adults. AB - Background Cardiovascular disease is the leading cause of death for adults in Australia. In recent years there has been a shift in health service delivery from institutional to community-based care for chronic conditions, including cardiovascular disease. The general practice setting is seen to offer greater flexibility, higher levels of efficiency and more client focused healthcare delivery than is possible in the acute care sector. It has been suggested that practice nurses represent a useful adjunct to current models of cardiovascular disease management. To date, significant descriptive research has been conducted exploring the demographics, roles, educational needs and issues facing practice nurses. However, there is a need to evaluate the effectiveness of practice nurse interventions in terms of patient outcomes, clinician satisfaction and cost effectiveness. Objectives This review seeks to present the best available evidence regarding the efficacy of general practice nurse interventions for cardiac risk factor reduction in healthy adults, as well as those with established cardiovascular disease or known cardiac risk factors. Search Strategy A systematic literature search was performed using Medline (1966 - 2005), CINAHL (1982 -2005), Cochrane Controlled Trials Register (Issue 4, 2005) and the Joanna Briggs Institute Evidence Library. In addition, the reference lists of retrieved papers, conference proceedings and the Internet, were scrutinised for additional trials. Selection Criteria This review considered any English language randomised trials that investigated interventions conducted by the practice nurse for cardiovascular disease management or reduction of cardiac risk factors. Interventions conducted by specialist cardiac nurses in general practice were excluded. Outcomes measured included blood pressure, smoking cessation, total cholesterol, exercise, body weight/body mass index and cost-effectiveness. Results Eighteen trials, reported in 33 papers, were included in the review. Ten trials investigated multifaceted interventions, while the remaining eight trials reported targeted interventions. Of the trials that reported multifaceted interventions, three trials investigated risk reduction in those with established cardiovascular disease, four trials focused on those with known cardiovascular disease risk factors and three trials included the general community. The eight trials which examined the efficacy of targeted interventions focused upon dietary intake (two trials), smoking cessation (three trials), weight reduction (one trial) and physical activity (two trials). The effect of both the multifaceted and targeted interventions on patient outcomes was variable. However, both the multifaceted and targeted interventions demonstrated similar outcome trends for specific variables. Improvements were demonstrated by most studies in blood pressure, cholesterol level, dietary intake and physical activity. The variation in outcome measures and contradictory findings between some studies makes it difficult to draw definitive conclusions. Conclusions While interventions to reduce cardiovascular disease risk factors have produced variable results, they offer significant potential to assist patients in modifying their personal risk profile and should be developed. The public health importance of these changes is dependant upon the sustainability of the change and its effect on the health outcomes of these individuals. Further well-designed research is required to establish the effectiveness of practice nurse interventions for cardiovascular disease management and risk factor reduction in terms of patient outcomes and cost-effectiveness. PMID- 21631793 TI - Meta-analysis comparing clinical effectiveness of drug-eluting stents, bare metal stents and coronary artery bypass surgery. AB - Objective To compare clinical outcomes among patients receiving drug-eluting stents, bare metal stents, or coronary artery bypass grafting surgery (CABG) to treat coronary artery disease. Data sources Randomised controlled trials were systematically selected from electronic database for head-to-head comparisons. The results from these head-to-head comparisons were used for an adjusted indirect comparison. Methods Published randomised controlled trials were reviewed for outcome data in patients treated for coronary artery disease with drug-eluting stents, bare metal stents, or CABG. Head-to-head comparisons were conducted for drug-eluting stents versus bare metal stents and for CABG versus bare metal stents. Adjusted indirect comparison was used to compare drug-eluting stents and CABG. Mid-term clinical outcomes (range: 6-12 months) were investigated and included rates of mortality, myocardial infarction, thrombosis, target lesion revascularisation, target vessel revascularisation, restenosis and major adverse cardiac events. Results Systematic literature search identified 23 randomised controlled trials (15 for drug-eluting stents vs. bare metal stents, 8 for CABG vs. bare metal stents). Head-to-head comparisons for both single and multiple vessel disease demonstrated that compared with bare metal stents, drug eluting stents had better outcomes for target lesion revascularisation, target vessel revascularisation, restenosis and major adverse cardiac events. Except target lesion revascularisation, data were similarly favourable for CABG when compared with bare metal stents. Adjusted indirect comparison between drug eluting stents and CABG in single vessel disease failed to detect significant differences in any of the measured outcomes. Multiple vessel disease data analysis demonstrated that target vessel revascularisation (odds ratio 3.41 [95% CI 2.29-5.08]) and major adverse cardiac events (1.89 [1.28-2.79]) were superior to drug-eluting stents in patients undergoing CABG. Conclusions Drug-eluting stents and CABG were superior to bare metal stents in terms of target lesion revascularisation (drug-eluting stents only), target vessel revascularisation, restenosis and major adverse cardiac events. There was no difference in clinical outcomes when comparing CABG and drug-eluting stents in patients with single vessel disease, and CABG may be superior to drug-eluting stents for target vessel revascularisation and major adverse cardiac events in patients with multiple vessel disease. However, results may vary between subpopulations with different clinical or socioeconomic differences. PMID- 21631794 TI - Water for wound cleansing. AB - Background Various solutions have been recommended for cleansing wounds, however, normal saline is favoured as it is an isotonic solution and is less likely to interfere with the normal healing process. Tap water is commonly used in the community for cleansing wounds because it is easily accessible, efficient and cost-effective; however, there is an unresolved debate about its use. Objectives The objective of this review was to assess the effects of water compared with other solutions for wound cleansing. Search strategy Randomised and quasi-randomised controlled trials were identified by electronic searches of Cochrane Wounds Group Specialised Register (June 2004), MEDLINE (1996-2004), EMBASE (1980-2004), CINAHL (1982-2004), and the Cochrane Controlled Trials Register (Issue 2, 2004). Primary authors, company representatives and content experts were contacted to identify eligible studies. Reference lists from included trials were also searched. Selection criteria Randomised and quasi randomised controlled trials that compared the use of water with other solutions for wound cleansing were eligible for inclusion. Additional criteria were outcomes that included objective or subjective measures of wound infection or healing. Data collection and analysis Trial selection, data extraction and quality assessment were carried out independently by two reviewers and checked by a third reviewer. Differences in opinion were settled by discussion. Some data were pooled using a fixed effects model. Main results Five trials were identified that compared rates of infection and healing in wounds cleansed with water and normal saline; three compared cleansing with no cleansing and one compared cleansing with procaine spirit with water. There were no standard criteria for the assessment of wound infection across the trials which limited the ability to pool the data. The major comparisons were water versus normal saline, and tap water versus no cleansing. For chronic wounds, the relative risk of developing an infection when cleansed with tap water compared with normal saline was 0.16 (95% CI 0.01-2.96). Use of tap water to cleanse acute wounds in adults was associated with a lower rate of infection than saline (RR 0.55, 95% CI 0.31-0.97). The use of tap water to cleanse acute wounds in children was not associated with a statistically significant difference in infection, when compared with saline (RR 1.07, 95% CI 0.43-2.64). Similarly, no statistically significant differences in infection rates were seen when wounds were cleansed with tap water versus not cleansed at all (RR 0.81, 95% CI 0.31-2.12). Likewise, there was no difference in the infection rate in episiotomy wounds cleansed with water or procaine spirit. The use of isotonic saline, distilled water and boiled water for cleansing open fractures also did not demonstrate a statistically significant difference in the number of fractures that were infected. Conclusions Although the evidence is limited one trial has suggested that the use of tap water to cleanse acute wounds reduces the infection rate and other trials conclude that there is no difference in the infection and healing rates between wounds that were not cleansed and those cleansed with tap water and other solutions. In the absence of drinkable tap water, boiled and cooled water as well as distilled water can be used as cleansing agents. PMID- 21631795 TI - Nursing administration of medication via enteral tubes in adults: a systematic review. AB - Background Enteral tubes are frequently inserted as part of medical treatment in a wide range of patient situations. Patients with an enteral tube are cared for by nurses in a variety of settings, including general and specialised acute care areas, aged care facilities and at home. Regardless of the setting, nurses have the primary responsibility for administering medication through enteral tubes. Medication administration via an enteral tube is a reasonably common nursing intervention that entails a number of skills, including preparing the medication, verifying the tube position, flushing the tube and assessing for potential complications. If medications are not given effectively through an enteral tube, harmful consequences may result leading to increased morbidity, for example, tube occlusion, diarrhoea and aspiration pneumonia. There are resultant costs for the health-care system related to possible increased length of stay and increased use of equipment. Presently what is considered to be best practice to give medications through enteral tubes is unknown. Objectives The objective of this systematic review was to determine the best available evidence on which nursing interventions are effective in minimising the complications associated with the administration of medications via enteral tubes in adults. Nursing interventions and considerations related to medication administration included form of medication, verifying tube placement before administration, methods used to give medication, methods used to flush tubes, maintenance of tube patency and specific practices to prevent possible complications related to the administration of enteral medications. Search strategy The following databases were searched for literature reported in English only: CINAHL, MEDLINE, The Cochrane Library, Current Contents/All Editions, EMBASE, Australasian Medical Index and PsychINFO. There was no date restriction applied. In addition, the reference lists of all included studies were scrutinised for other potentially relevant studies. Selection criteria Systematic reviews of randomised controlled trials (RCTs) and RCTs that compared the effectiveness of nursing interventions and considerations used in the administration of medications via enteral tubes. Other research methods, such as non-randomised controlled trials, longitudinal studies, cohort and case control studies, were also included. Exclusion criteria included studies investigating drug-nutrient interactions or the bioavailability of specific medications. Data collection and analysis Initial consideration of potential relevance to the review was carried out by the primary author (NP). Two reviewers independently assessed study eligibility for inclusion. A meta-analysis could not be undertaken, as there were no comparable RCTs identified. All data were presented in a narrative summary. Results There is very limited evidence regarding the effectiveness of nursing interventions in minimising the complications associated with enteral tube medication administration in adults. The review highlights a lack of high quality research on many important nursing issues relating to enteral medication administration. There is huge scope for further research. Some of the evidence that was identified included that nurses should consider the use of liquid form medications as there may be fewer tube occlusions than with solid forms in nasoenteral tubes and silicone percutaneous endoscopic gastronomy tubes. Nurses may need to consider the sorbitol content of some liquid medications, for example, elixirs, as diarrhoea has been attributed to the sorbitol content of the elixir, not the drug itself. In addition, the use of 30 mL of water for irrigation when administering medications or flushing small diameter nasoenteral tubes may reduce the number of tube occlusions. PMID- 21631796 TI - An evaluation of the implementation of a best practice guideline on tracheal suctioning in intensive care units. AB - Aim To minimise suctioning-induced complications in intensive care patients, it is crucial that nurses are able to perform the procedure safely and act in accord with research-based recommendations. This paper reports the process of developing, disseminating and implementing the best practice guideline and an evaluation of the process and outcomes of care during and following its implementation in intensive care units. Methods The study was divided into four phases: (i) to develop the best practice guideline and plan strategies for its dissemination and implementation; (ii) to audit the current practice of nurses in the tracheal suctioning of patients in intensive care units with an artificial airway; (iii) to disseminate and implement the best practice guideline; and (iv) to evaluate the process as well as outcome of care following its implementation in intensive care units. Results The pretest results indicate that gaps exist between actual nursing practice and recommendations based on research evidence. Most nurses performed the skills in accord with the best practice guideline, with 65% nurses scoring above the 70% level. The post-test audit results show that, overall, nurses demonstrated a good endotracheal suctioning technique, with 96% scoring above 75%, indicating an overall improvement in compliance with the guideline. A statistically significant difference was found between the pretest (73%) and post-test (89%) compliance scores (t = -7.67, P < 0.005). Conclusions This implementation project highlights the importance of using a rigorous and systematic process to ensure the formal testing of an intervention. Some essential principles in implementing evidence are necessary, such as involving relevant staff and having a range of strategies and clear processes for implementation. PMID- 21631798 TI - Response to Critique of Cochrane systematic review of home-like setting for birth in the International Journal of Evidence-Based Healthcare. PMID- 21631797 TI - Critique of Cochrane systematic review of home-like setting for birth. AB - This paper is a critique of a Cochrane systematic review where the primary objective of the review was to evaluate the effects, on labour and birth outcomes, of care in a home-like birth setting compared with care in a conventional hospital labour ward. The unanticipated finding of that review was a trend towards higher perinatal mortality in the home-like setting group. Our critique focuses on one important question: is the Cochrane review of birth setting based on studies of sufficient scientific rigour to be able to establish whether or not there may be an added risk of perinatal mortality associated with intrapartum care in a home-like setting? In this commentary we critique two aspects of methodological quality in the studies underpinning the Cochrane review: the high levels of non-compliance with the planned treatment, and the low levels of 'treatment fidelity' even if subjects received the planned treatment. Of the 41 deaths in the home-like setting group 35 (85%) were not associated with intrapartum care in a home-like setting. We conclude that the Cochrane review of birth setting cannot be relied upon to make valid statements about whether there may be an added risk of perinatal mortality associated with experiencing intrapartum care in either setting. In the process of our analysis we uncovered what may be two broader problems for Cochrane systematic reviews. The Cochrane Reviewers' Handbook should be expanded to give specific guidance for researchers and reviewers about how to enhance or evaluate research quality in relation to non-compliance with assigned treatment and treatment fidelity. PMID- 21631799 TI - Rejoinder. PMID- 21631801 TI - Improving health and healthcare. PMID- 21631802 TI - Effectiveness of brief structured interventions on risk factor modification for patients with coronary heart disease: a systematic review. AB - Background The physical and psychosocial benefits of participation in cardiac rehabilitation following a coronary event have well been established. Despite these benefits there is strong evidence that participation in traditional cardiac rehabilitation programs remains low. Various models of cardiac rehabilitation have been implemented including the use of brief structured interventions to enable modification of coronary risk factors. Objectives The objective of this review was to determine the effect of brief structured interventions on risk factor modification in patients with coronary heart disease. Search strategy A literature search was performed using the following databases MEDLINE (1966 2006), CINAHL (1982-2006), EMBASE (1980-current) and up to the Cochrane Controlled Trials Register (Issue 2, 2006 of Cochrane Library). In addition, the reference lists of relevant trials and conference proceedings were also scrutinised. Company representatives, experts and investigators were contacted to elicit further information. Selection criteria All randomised and quasi randomised controlled trials that compared the effects of brief structured interventions on risk factor modification in patients with coronary heart disease were considered for inclusion in the review. Data collection and analysis Eligibility of the trials for inclusion in the review, details of eligible trials and the methodological quality of the trials were assessed independently by two reviewers. Relative risks for dichotomous data and a weighted mean difference for continuous data were calculated with 95% confidence intervals. Where synthesis was inappropriate, trials were considered separately. Main results Seventeen trials involving a total of 4725 participants were included in the final review: three trials compared the effects of brief structured interventions on diet modification; seven on smoking cessation; and seven on multiple risk factors. Two trials involving 76 patients compared brief structured intervention versus usual care for dietary modification. Although there was a tendency for more participants in the intervention arm to lose weight at the 12-week follow up and achieve target cholesterol levels at the 6-month follow up, these results were not statistically significant. Only one small trial involving 36 patients compared brief structured intervention and extensive intervention for dietary modification and demonstrated a significant reduction in the percentage of energy obtained from fat and saturated fat intake among participants receiving extensive intervention. However, no difference in fish, fruit and vegetable intake between the groups was evident. Six trials involving 2020 patients compared brief structured intervention versus usual care for smoking cessation. There was no difference in the smoking cessation rates at the 3- and 6-week follow up, however, there was evidence of a benefit of brief structured interventions for smoking cessation at the 3-, 6- and 12-month follow up. In the only trial that and compared brief structured intervention and extensive intervention for smoking cessation in 254 participants there was no clear difference of a likelihood of smoking cessation between the two groups. In the seven trials that compared brief structured intervention and usual care for multiple risk factor modification there was evidence of a benefit of the intervention on behavioural changes such as fat intake, weight loss and consequently on reduction in the body mass index, smoking cessation and physical activity among the participants. The findings concerning the effect on blood pressure, blood glucose levels and the lipid profile, however, remain inconclusive. Conclusions There is suggestive but inconclusive evidence from the trials of a benefit in the use of brief interventions for risk factor modification in patients with coronary heart disease. This review, however, supports the concept that brief interventions for patients with coronary heart disease can have beneficial effects on risk factor modification and consequently on progression of coronary heart disease. Further trials using larger sample sizes need to be undertaken to demonstrate the benefits of brief structured intervention targeted at the modification of single or multiple risk factors. PMID- 21631803 TI - Managing the wandering behaviour of people living in a residential aged care facility. AB - Background Wandering behaviour is frequently seen in older people with cognitive impairment. The prevalence of patients exhibiting wandering behaviour has been estimated to be 11.6% on traditional units and 52.7% on Alzheimer's units. Wandering is one of the core behavioural characteristics that impact on familial carers and is likely to influence the decision to place a family member in an aged care environment. Considering the possible risks associated with wandering behaviour, the successful identification and management of wandering is essential. Wandering is also a problem for caregivers in the institutionalised setting, in terms of containment, usually being addressed by securing the environment. There has been some research conducted to assist in the understanding and management of wandering behaviour; however, the findings have been diverse resulting in a level of confusion about the best approaches to take. Objectives This review aims to present the best available evidence on the management of wandering in older adults who reside in an aged care facility (both high and low care). Search strategy An extensive search of keywords contained in the title and abstract, and relevant MeSH headings and descriptor terms was performed on the following databases: MEDLINE, CINAHL, PsychINFO, AGELINE, Cochrane Library, Embase, APAIS Health, Current Contents, Dare, Dissertation Abstracts, Personal Communication, Social Science Index. Selection criteria Papers were selected if they focused on the treatment of wandering in an institutional setting. Some studies were not specifically examining wanderers over the age of 65 years as per the protocol requirements, but were included as it was felt that their findings could be applied to this age group. Data collection and analysis Study design and quality were tabulated and relative risks, odds ratios, mean differences and associated 95% confidence intervals were calculated from individual comparative studies containing count data where possible. All other data were presented in a narrative summary. Results Searches identified one care protocol, two systematic reviews and 24 other studies that satisfied the inclusion criteria. The following recommendations are divided into four categories of interventions (environmental, technology and safety, physical/psychosocial, and caregiving support and education) with only Level 1, 2 or 3 evidence presented. Environmental modifications Gridlines placed in front of doors or covering exit door doorknobs or panic bars may be effective at reducing exit-seeking behaviour (Level 3b). Technology and safety Mobile locator devices may be effective at enabling quick location of wandering residents (Level 3c). Physical/psychosocial interventions Implementation of a walking group or an exercise program may reduce the incidence of disruptive wandering behaviour (Level 3b). Use of air mat therapy may reduce wandering behaviour for at least 15 min post therapy (Level 2). Providing music sessions (and reading sessions) may keep residents from wandering during the period of the session (Level 3b). Caregiving support and education There is no evidence to support any interventions. Conclusions The majority of the available research for which the guidelines are based upon was derived from observational studies or expert opinion (Level of evidence 3 or 4). More rigorous research is required to demonstrate the efficacy of these recommendations. PMID- 21631804 TI - The effectiveness of non-surgical interventions in the treatment of Charcot foot. AB - Background Charcot neuropathic osteoarthropathy is commonly known as 'Charcot foot'. It is a serious foot complication of diabetes mellitus that can frequently lead to foot ulceration, gangrene, hospital admission and foot amputation. A multidisciplinary approach to the management of Charcot foot is taken involving medical and allied health professionals. The management approach may also differ between different countries. To date, there is no systematic review of the literature undertaken to identify the clinical effectiveness of non-operative interventions in the treatment of acute Charcot foot. Objective The objective of this review was to identify the effectiveness of non-surgical interventions with reducing lesions, ulceration, the rate of surgical intervention, reducing hospital admissions and improve the quality of life of subjects with Charcot foot. Search strategy A comprehensive search strategy was undertaken on databases available from University of South Australia from their inception to November 2006. Selection criteria Randomised controlled trials or clinical controlled trials were primarily sought. Critical appraisal of study quality and data extraction was undertaken using Joanna Briggs Institute instruments. Review Manager software was used to calculate comparative statistics. Results This review identified 11 trials and five trials were included in the review. Three trials involved the use of bisphosphonate, a pharmacological agent. Two experimental treatments were also included, evaluating palliative radiology and magnetic fields. No trials were found using immobilisation and off-loading interventions for acute Charcot foot. The overall methodological quality score of the five studies was moderate. Owing to heterogeneous data, meta-analysis could not be performed. The trials did not report on reducing lesions, ulceration, rate of surgical intervention, hospital admissions and the quality of life of subjects with Charcot foot. The trials evaluating bisphosphonates reported greater reduction in foot temperature and disease activity for intervention subjects compared with controls. Another outcome of this review indicated additional beneficial effects of bisphosphonates in reducing pain and discomfort. The trial evaluating palliative radiotherapy found no difference between groups on any outcome. A significant reduction in the amount of deformity and reduced healing time to consolidation was found after treatment in the group receiving magnetic therapy treatment. Discussion There is a lack of clinical trials evaluating the effectiveness of non-operative interventions for the management of Charcot foot (immobilisation, removable cast walkers, advice/dispensing of footwear and prescribing of orthotics). Bisphosphonates may be useful adjuncts to standard management of Charcot foot by improved healing demonstrated by a reduction in disease activity indicated by skin temperature and bone destruction. Magnetic therapy may reduce deformity, joint destruction and improve mobility. Conclusion There is a lack of evidence supporting the use of pharmacological or non-surgical interventions with reducing lesions, ulceration, rate of surgical intervention, hospital admissions and improving the quality of life of subjects with Charcot foot. Bisphosphonates may improve the healing of Charcot foot by reducing skin temperature and disease activity of Charcot foot, when applied in addition to standard interventions to control the position and shape of the foot. PMID- 21631805 TI - Advance care planning for residents in aged care facilities: what is best practice and how can evidence-based guidelines be implemented? AB - Background Advance care planning in a residential care setting aims to assist residents to make decisions about future healthcare and to improve end-of-life care through medical and care staff knowing and respecting the wishes of the resident. The process enables individuals and others who are important to them, to reflect on what is important to the resident including their beliefs/values and preferences about care when they are dying. This paper describes a project conducted as part of the Joanna Briggs Institute Clinical Aged Care Fellowship Program implemented at the Manningham Centre in metropolitan Melbourne in a unit providing services for 46 low and high care residents. Objectives The objectives of the study were to document implementation of best practice in advance care planning in a residential aged care facility using a cycle of audit, feedback and re-audit cycle audit with a clinical audit software program, the Practical Application of Clinical Evidence System. The evidence-based guidelines found in 'Guidelines for a Palliative Approach in Residential Aged Care' were used to inform the process of clinical practice review and to develop a program to implement advance care planning. Results The pre-implementation audit results showed that advance care planning practice was not based on high level evidence as initial compliance with five audit criteria was 0%. The barriers to implementation that became apparent during the feedback stage included the challenge of creating a culture where advance care planning policy, protocols and guidelines could be implemented, and advance care planning discussions held, by adequately prepared health professionals and carers. Opportunities were made to equip the resident to discuss their wishes with family, friends and healthcare staff. Some residents made the decision to take steps to formally document those wishes and/or appoint a Medical Enduring Power of Attorney to act on behalf of the resident when they are unable to communicate wishes. The post-implementation audit showed a clear improvement as compliance ranged from 15-100% for the five audit criteria. Strong leadership by the project team was effective in engaging staff in this quality improvement program. Conclusion The outcomes of the project were extremely positive and demonstrate a genuine improvement in practice. All audit criteria indicate that the Manningham Centre is now positively working towards improved practice based on the best available evidence. It is hoped that as the expertise developed during this project is shared, other areas of gerontological practice will be similarly improved and more facilities caring for the older person will embrace evidence-based practice. PMID- 21631806 TI - Reducing and/or minimising physical restraint in a high care, rural aged care facility. AB - Background This report takes you through a journey of implementing evidence based best practice guidelines in relation to physical restraint in an aged care facility. It describes the processes involved in making changes to an existing system, introducing evidence-based guidelines and collaboratively achieving compliance with best practice. The project formed part of a combined initiative between the Joanna Briggs Institute and the Commonwealth Department of Health and Ageing, called the Aged Care Clinical Fellowship Program. The Joanna Briggs Institute Practical Application of Clinical Evidence System, an online audit based tool that includes an action research process Getting Research Into Practice was used to facilitate audit data collection and analysis. This report demonstrates that effective clinical leadership and evidence-based research, combined with a strategy of audit, feedback and re-audit, can become an effective change agent to improve clinical practice in residential aged care facilities. Aims/objectives The aims of this project were to increase staff knowledge and awareness of restraints, improve practices in restraint assessment and usage, reduce or minimise the amount and type of restraints used and ensure the least restrictive device possible was utilised in a rural aged care facility. Methods A system of audit, feedback and re-audit was performed using the Joanna Briggs Institute Practical Application of Clinical Evidence System software. This was completed in a series of stages over a period of 6 months. The first stage was to apply defining characteristics to each of the evidence-based audit criteria to determine compliance. A team of staff was set up to assist with the project. An initial audit was conducted, followed by a situational analysis of the findings. From this an action plan for improvement using Getting Research Into Practice was developed. The action plan was then implemented and the criteria re-audited and reviewed. Results The initial audit confirmed non-compliance as expected in two criteria. All criteria showed improvement in the re-audit. Those criteria with the least compliance in the initial audit showed the most improvement in the re audit. Conclusion Effective change processes can be achieved when there is an identified reason or need for change, and when staff are aware of that need. With clinical leadership and education attitudes, fears and myths can be dispelled and improved performance will come out. PMID- 21631807 TI - Effectiveness of education in evidence-based healthcare: the current state of outcome assessments and a framework for future evaluations. AB - Background A discipline which critically looks at the evidence for practice should itself be critically examined. Credible evidence for the effectiveness of training in evidence-based healthcare (EBHC) is essential. We attempted to summarise the current knowledge on evaluating the effectiveness of training in EBHC while identifying the gaps. Methods A working group of EBHC teachers developed a conceptual framework of key areas of EBHC teaching and practice in need of evidence mapped to appropriate methods and outcomes. A literature search was conducted to review the current state of research in these key areas. Studies of training interventions that evaluated effectiveness by considering learner, patient or health system outcomes in terms of knowledge, skills, attitude, judgement, competence, decision-making, patient satisfaction, quality of life, clinical indicators or cost were included. There was no language restriction. Results Of 55 articles reviewed, 15 met the inclusion criteria: six systematic reviews, three randomised controlled trials and six before-after studies. We found weak indications that undergraduate training in EBHC improves knowledge but not skills, and that clinically integrated postgraduate teaching improves both knowledge and skills. Two randomised controlled trials reported no impact on attitudes or behaviour. One before-after study found a positive impact on decision-making, while another suggested change in learners' behaviour and improved patient outcome. We found no studies assessing the impact of EBHC training on patient satisfaction, health-related quality of life, cost or population-level indicators of health. Conclusions Literature evaluating the effectiveness of training in EBHC has focused on short-term acquisition of knowledge and skills. Evaluation designs were methodologically weak, controlled trials appeared inadequately powered and systematic reviews could not provide conclusive evidence owing to weakness of study designs. PMID- 21631808 TI - Supporting evidence-based service delivery and organisation: a comparison of an emergent realistic appraisal technique with a standard qualitative critical appraisal tool. AB - A major component of current policies aimed at promoting the delivery of an effective health service is to ensure that practitioners base their practice on research findings. Considerable attention has been focussed on the research community and developing appropriate methods and tools to support these objectives at the national and international level. The appraisal needs of individual practitioners, who might wish to inform local service delivery and organisation and who are often not methodological experts, has been hitherto ignored. This paper reports on the application and testing of a technique for the appraisal of evaluation studies, which is being developed for use by practitioners to inform local service delivery. This technique has previously been applied to a quantitative evaluation. In this paper we report on its application to the appraisal of a qualitative paper. Our aim is to compare it with a standard tool to establish which generates more meaningful information for use by practitioners for the purposes of informing service delivery and organisation. The emergent technique successfully extracted relevant methodological and contextual information, with developmental issues being identified. PMID- 21631809 TI - Context or quality, are they mutually exclusive? A response to Satherley, Allen and Lyne. PMID- 21631810 TI - Context or quality are they mutually exclusive? A rejoinder. PMID- 21631811 TI - The uncertainty and incongruity of evidence-based healthcare. PMID- 21631812 TI - Clinical effectiveness of barrier preparations in the prevention and treatment of nappy dermatitis in infants and preschool children of nappy age. AB - Nappy dermatitis is a broad term used to describe an acute inflammatory reaction of the skin in the nappy area because of irritation from urine, faeces, moisture or friction. The prevalence is estimated to be between 7% and 35% in infants. Regular application of a barrier preparation at every nappy change may be a valuable component of nappy dermatitis prevention and/or treatment. PMID- 21631813 TI - Effects of music listening on adult patients' pre-procedural state anxiety in hospital. AB - EXECUTIVE SUMMARY: BACKGROUND: Anticipation of an invasive procedure in hospital is likely to provoke feelings of anxiety and stress in patients. An unfamiliar environment, loss of control, perceived or actual physical risk, dependence on strangers and separation from friends and family are all factors that can contribute to the development of such feelings. Recently, there has been considerable interest in the anxiolytic potential of music listening in a variety of clinical settings, yet thus far, little is known about the impact of music listening on the pre-procedural patient population. A systematic review of all literature to date was indicated to improve understanding of outcomes and impact of music listening on pre-procedural anxiety, thus helping nurses decide whether or not to incorporate music listening into practice and to highlight a need, or otherwise, for a related primary research agenda. OBJECTIVE: The objective of this review was to determine the best available evidence on the effectiveness of music listening in reducing adult hospital patients' pre-procedural state anxiety. INCLUSION CRITERIA: Types of studies This review included randomised controlled trials and quasi-experimental research designs that examined the efficacy of music listening in reducing state anxiety among pre-procedural hospital patients published between January 1985 and February 2006. The search was limited to publications after 1985 to coincide with the increasing interest and use of complementary therapies within health care during the 1980s and 1990s. Types of participants Participants of interest to the review were adult day patients, ambulatory patients and inpatients who were about to undergo any type of clinical procedure. Types of intervention The review focused on studies that investigated pre-procedural music listening employed and prescribed as a potentially therapeutic activity. It excluded any other form of music therapy. Types of outcome measures The primary outcome measures examined were alterations in state anxiety and a variety of physiological variables such as blood pressure and respiration and heart rates. SEARCH STRATEGY: A search for published and unpublished literature between January 1985 and February 2006 was conducted using all major electronic databases. A three-step search strategy was devised which consisted of using high-precision MeSH terminology and keywords to ensure that all material relevant to the review was captured. CRITICAL APPRAISAL: The methodological quality of included studies was assessed by two reviewers, who appraised each study independently, using the standard Joanna Briggs Institute (JBI) critical appraisal tools. DATA EXTRACTION AND SYNTHESIS: Data were extracted from the studies that were identified as meeting the criteria for methodological quality using a data-extraction tool developed for the review. Studies were grouped by outcome measure and summarised using tabular and narrative formats. RESULTS: The review demonstrated that state anxiety is defined and measured using both psychological and physiological parameters. Music listening had a consistently positive and statistically significant effect on reducing psychological parameters of pre-procedural state anxiety. However, the results from the measurement of various pre-procedural physiological parameters failed to reveal any consistent positive changes in patients who had listened to music. This calls into question the adequacy of the theories in this area which link anxiety and the automated and central nervous systems and the effect that music listening may have on these processes and physiological responses. CONCLUSIONS/IMPLICATIONS FOR PRACTICE: 1 In order to reduce anxiety, it is likely that patients will benefit psychologically from having the opportunity to listen to music in the immediate pre-procedural period. 2 Patients do not appear to experience any alteration in physiological status as a result of listening to music. 3 Further research is indicated in order to replicate existing studies, to strengthen the evidence to support such interventions and to establish intervention parameters. 4 Further research is needed analysing the physiological mechanisms by which music listening is believed to reduce state anxiety and the contribution of the automated and other nervous systems to this reduction. PMID- 21631814 TI - Role of MRSA reservoirs in the acute care setting. AB - Background Nosocomial infection remains the most common complication of hospitalisation. Despite infection control efforts, nosocomial methicillin resistant Staphylococcus aureus (MRSA) transmission continues to rise. The associated costs of increased hospital stay and patient mortality cause considerable burden to the health system. Objectives This review sought to evaluate the role of reservoirs, particularly the environment and equipment commonly found in the clinical area, in the transmission of MRSA within the acute hospital. This review updates a review previously completed by the authors and published by the Joanna Briggs Institute (2002). Search strategy A systematic search for relevant published or unpublished literature was undertaken using electronic databases, the reference lists of retrieved papers and the Internet. This extended the search published in the original review. Databases searched included Medline (1966-August Week 1 2005), CINAHL (1982-August Week 1 2005), EMBASE (1996-Week 33), as well as the Cochrane Library (Issue 3, 2005) and the Joanna Briggs Institute Evidence Library (August 2005). Selection criteria All research reports published between 1990 and August 2005 in the English language that focused on the role of the environment and equipment commonly found in the clinical area on the nosocomial MRSA transmission in adult, paediatric or neonatal acute care settings were considered. Data collection and analysis Two reviewers assessed each paper against the inclusion criteria and a validated quality scale. Studies that scored less than the mean quality score were excluded from the review. Data extraction was undertaken using a tool designed specifically for this review. Statistical comparisons of findings were not possible, so findings are presented in a narrative form. Results Forty-two papers met the review inclusion criteria, of which 18 obtained a quality score above the threshold and are included in this review. Seven studies reported general investigations of MRSA in the clinical environment and 11 studies explored specific environmental aspects. All studies used exploratory, descriptive or comparative designs. The evidence suggests that MRSA strains within the environment often match those found in patients within that environment. MRSA can be found in the air around MRSA colonised or infected patients. The degree of airborne contamination is significantly increased by activities that promote airflow. Although the site of MRSA colonisation or infection can influence the degree of environmental contamination, these data are inconsistent. Therefore, there is limited evidence for tailoring infection control interventions based on the sites of MRSA colonisation or infection. The evidence suggests that the type of materials used in clinical equipment can influence the effectiveness of cleaning techniques. Current routine cleaning practices, including conventional terminal cleaning, do not necessarily effectively eradicate MRSA from the environment. This review demonstrates that there is a link between the environment and hospital equipment and the transmission of MRSA within the acute hospital setting. Further well-designed research is urgently required to explore the efficacy of specific cleaning and decontamination methods, staff compliance with infection control practices and the range of factors that affect the incidence of MRSA contamination of the environment and equipment commonly found in the clinical area. PMID- 21631815 TI - How has the impact of 'care pathway technologies' on service integration in stroke care been measured and what is the strength of the evidence to support their effectiveness in this respect? AB - EXECUTIVE SUMMARY: BACKGROUND: Across the developed world, we are witnessing an increasing emphasis on the need for more closely coordinated forms of health and social care provision. Integrated care pathways (ICPs) have emerged as a response to this aspiration and are believed by many to address the factors which contribute to service integration. ICPs map out a patient's journey, providing coordination of services for users. They aim to have: 'the right people, doing the right things, in the right order, at the right time, in the right place, with the right outcome'. The value for ICPs in supporting the delivery of care across organisational boundaries, providing greater consistency in practice, improving service continuity and increasing collaboration has been advocated by many. However, there is little evidence to support their use, and the need for systematic evaluations in order to measure their effectiveness has been widely identified. A recent Cochrane review assessed the effects of ICPs on functional outcome, process of care, quality of life and hospitalisation costs of inpatients with acute stroke, but did not specifically focus on service integration or its derivatives. To the best of our knowledge, no such systematic review of the literature exists. OBJECTIVES: * To systematically review all high-quality studies which have evaluated the impact of care pathway technologies on 'service integration' and its derivatives in stroke care * To examine how elements of service integration are defined in such studies * To examine the type of evidence utilised to measure service integration * To analyse the weight of evidence used to support claims about the effectiveness of ICPs on improving service integration * To produce recommendations for ICP developers, users and evaluators. INCLUSION CRITERIA: Types of participants The review focused on the care of adult patients who had suffered a stroke. It included the full spectrum of services - acute care, rehabilitation and long-term support - in hospital and community settings. Types of intervention(s)/phenomena of interest Integrated care pathways were the intervention of interest, defined for the purpose of this review as 'a multidisciplinary tool to improve the quality and efficiency of evidence based care and is used as a communication tool between professionals to manage and standardise the outcome orientated care' Here 'multidisciplinary' is taken to refer to the involvement of two or more disciplines. Types of outcomes 'Service integration' was the outcome of interest however, this was defined and measured in the selected studies. Types of studies This review was concerned with how 'service integration' was defined in evaluations of ICPs; the type of evidence utilised in measuring the impact of the intervention and the weight of evidence to support the effectiveness of care pathway technologies on 'service integration'. Studies that made an explicit link between ICPs and service integration were included in the review. Evidence generated from randomised controlled trials, quasi-experimental, qualitative and health economics research was sought. The search was limited to publications after 1980, coinciding with the emergence of ICPs in the healthcare context. Assessment for inclusion of foreign papers was based on the English-language abstract, where available. These were included only if an English translation was available. EXCLUSION CRITERIA: This review excluded studies that: * focused only on a single aspect of stroke care (e.g. dysphasia) * evaluated ICPs as part of a wider program of service development * did not make an explicit link between ICPs and service integration * did not meet the definition of ICP utilised for the purposes of the review * focused exclusively on the outcomes of variance analysis SEARCH STRATEGY: In order to avoid replication, the Joanna Briggs Institute for Evidence Based Nursing and Midwifery Database and the Cochrane Library were searched to establish that no systematic reviews existed and none were in progress. A three stage search strategy was then used to identify both published and unpublished studies (see Appendix III). DATA COLLECTION: Our search strategy located 2123 papers, of which 39 were retrieved for further evaluation. We critically appraised seven papers, representing five studies. These were all evaluation studies and, as is typical in this field, comprised a range of study designs and data collection methods. Owing to the diversity of the study types included in the review, we developed a single-appraisal checklist and data-extraction tool which could be applied to all research designs.(32) The tool drew on the Joanna Briggs Institute (JBI) appraisal checklists for experimental studies and interpretive and critical research, and also incorporated specific information and issues which were relevant for our purposes (see Appendix VI). This extends the thinking outlined in Lyne et al.(31) in which, drawing on Campbell and Stanley's classic paper, the case is made for developing an appraisal tool which is applicable to all types of evaluation, irrespective of study design. In assessing the quality of the papers, we were sympathetic to the methodological challenges of evaluating complex interventions such as ICPs. We were also cognisant of the very real constraints in which service evaluations are frequently undertaken in healthcare contexts. In accordance with the aims of this particular review, we have included studies, which are methodologically weaker than is typical of many systematic reviews because, in our view, in the absence of stronger evidence, they yield useful information. DATA SYNTHESIS: Given the heterogeneity of the included studies, meta-analysis and/or qualitative synthesis was not possible. A narrative summary of the study findings is presented. RESULTS: 1 ICPs can be effective in ensuring that patients receive relevant clinical interventions and/or assessments in a timely manner, although these improvements may reflect better documentation rather than actual changes in practice. 2 ICPs can be effective in improving the documentation of rehabilitation goals, documentation of communication with patients, carers (diagnosis, prognosis and follow-up arrangements) and documentation of notification of primary care physicians of discharge. However, this can create additional burdens of work for staff. 3 Early studies of ICP-managed care in the acute stroke context have demonstrated reduced length of stay without any associated adverse effects on discharge destination, morbidity or mortality. These effects do not reach statistical significance, however, and may reflect wider changes in service provision and a general trend towards reduced length of hospital stay. While later studies in the acute and rehabilitation contexts do not reveal any significant reduction in length of stay, they do report greater documented use of certain clinical interventions and assessments, suggesting that ICPs can be effective in mobilising hospital resources around the patient. 4 ICPs implemented in the context of acute stroke care can be effective in reducing the occurrence of urinary tract infections, although we do not know whether this can be attributed to improved service integration. 5 ICP management in stroke rehabilitation may not be flexible enough to meet diverse patient needs and can result in insufficient attention to higher-level functioning and carer needs influencing perceptions of quality of life. 6 ICP management may assist in clarifying role boundaries and a shared understanding of the work, but this can result in some members of the disciplinary team perceiving that their contribution is not appropriately reflected in the documentation. 7 There is some evidence that ICPs may be effective in changing professional behaviours in the desired direction where there is scope for improvement, but in situations in which multidisciplinary working is effective, their positive effects may be limited. Furthermore, it is far from clear what the active ingredients of ICPs actually are. Kwan et al. suggest that it was the process of ICP development that had most impact on behaviours rather than the use of the artefact per se.(20) 8 None of the studies assessed the balance of costs and benefits of ICP use. Therefore, we do not know whether the costs of ICP development and implementation are justified by any of the reported benefits. CONCLUSIONS: Implications for practice There is some evidence that ICPs may support certain elements of service integration in the context of stroke care. This seems to be as a result of their ability to support the timely implementation of clinical interventions and the mobilisation of resources around the patient without incurring additional increases in length of stay. ICPs appear to be most successful in improving service coordination in the acute stroke context where patient care trajectories are predictable. Their value in the context of rehabilitation settings in which recovery pathways are more variable is less clear. There is some evidence that ICPs may be effective in bringing about behavioural changes in contexts where deficiencies in service provision have been identified. Their value in contexts where inter-professional working is well established is less clear. While earlier before and after studies show a reduction in length of stay in ICP-managed care, this may reflect wider healthcare trends, and the failure of later studies to demonstrate further reductions suggests that there may be limits as to how far this can continue to be reduced. There is some evidence to suggest that ICPs bring about improvements in documentation, but we do not know how far documented practice reflects actual practice. It is unclear how ICPs have their effects and the relative importance of the process of development and the artefact in use. (ABSTRACT TRUNCATED) PMID- 21631816 TI - Joanna Briggs Collaboration Aged Care Fellowship Project: implementing a smoking cessation program in a young, frail aged residential care facility. AB - Background The subject site (Ian George Court) caters for clients from a socially disadvantaged background. All clients have been homeless or at risk of homelessness and have a history of alcohol and substance abuse often linked to mental health issues. This project was developed to examine if the site provided best practice in the promotion of smoking cessation. Objectives The first objective of this project was to improve client knowledge to make informed choice about smoking cessation, ensuring that client advice was given in line with best available evidence and assist the client in accessing community programs. The second objective was to fully review the current assessment tool used in relation to gathering baseline data about smoking habits and act on the information provided. Search strategy The search strategy sought to find published studies and papers. An initial limited search of MEDLINE and CINAHL was undertaken followed by an analysis of the text words contained in the title and abstract. A second extensive search was then undertaken using all identified keywords. Conclusion A smoking assessment tool was developed and is now in use across all Anglicare sites in South Australia. This provides staff with consistent baseline information and offers evidence-based health care in a package format to aid clients in smoking cessation. PMID- 21631817 TI - Engaging consumers in safety and quality at Royal Adelaide hospital. AB - Objective The objective of this study was to elicit barriers and enablers of safe, high-quality care as identified by consumers, and to position consumers as 'possessors' of valuable knowledge related to systems and practices (as they had experienced these directly) rather than the receivers of knowledge and information. The central aim was to develop recommendations for consumer input into quality improvement, generated from the analysis of narrative accounts of their experiences. Methods The four-phase methodology adopted for this project involved the development of quality improvement strategies as identified (phase one) and validated (phase two) by consumers through the conduct of discovery interviews with 30 consumers over the age of 18 years who had experienced an adverse event. Clinicians and quality managers were then provided with an opportunity to validate the strategies identified through participation in a focus group (phase three). All data collected through discovery interviews and focus groups were transcribed and entered into the Joanna Briggs Institute Qualitative Assessment and Review Manager for analysis. The final phase of the study involved integrating this process of consumer involvement and of identified improvement strategies into the quality improvement program of Royal Adelaide Hospital. Results A total of 28 findings were entered into the Joanna Briggs Institute Qualitative Assessment and Review Manager for analysis. The process of meta-synthesis embodied in these programs involves the aggregation or synthesis of findings or conclusions. Six categories and four syntheses were derived through this process with key themes relating to assessment and prevention strategies, a necessity for improved education and communication, the hospital environment and the potential life impact that the experience of an adverse event may have. Conclusion Consumers identified a number of strategies that could contribute to improved safety and clinical outcomes in hospital and a reduction in adverse events. This current study provides a solid foundation upon which future research may be conducted. PMID- 21631818 TI - The gap between discovery and practice implementation in evidence-based practice: is practice-based evidence a solution? PMID- 21631819 TI - Effectiveness of interventions that assist caregivers to support people with dementia living in the community: a systematic review. AB - Objectives The objective of this review was to assess the effectiveness of interventions that assist caregivers to provide support for people living with dementia in the community. Inclusion criteria Types of participants Adult caregivers who provide support for people with dementia living in the community (non-institutional care). Types of interventions Interventions designed to support caregivers in their role such as skills training, education to assist in caring for a person living with dementia and support groups/programs. Interventions of formal approaches to care designed to support caregivers in their role, care planning, case management and specially designated members of the healthcare team - for example dementia nurse specialist or volunteers trained in caring for someone with dementia. Types of studies This review considered any meta-analyses, systematic reviews, randomised control trials, quasi-experimental studies, cohort studies, case control studies and observational studies without control groups that addressed the effectiveness of interventions that assist caregivers to provide support for people living with dementia in the community. Search strategy The search sought to identify published studies from 2000 to 2005 through the use of electronic databases. Only studies in English were considered for inclusion. The initial search was conducted of the databases, CINAHL, MEDLINE and PsychINFO using search strategies adapted from the Cochrane Dementia and Cognitive Improvement Group. A second more extensive search was then conducted using the appropriate Medical Subject Headings (MeSH) and keywords for other available databases. Finally, hand searching of reference lists of articles retrieved and of core dementia, geriatric and psycho geriatric journals was undertaken. Assessment of quality Methodological quality of each of the articles was assessed by two independent reviewers using appraisal checklist developed by the Joanna Briggs Institute and based on the work of the Cochrane Collaboration and Centre for Reviews and Dissemination. Data collection and analysis Standardised mean differences or weighted mean differences and their 95% confidence intervals were calculated for each included study reported in the meta analysis. Results from comparable groups of studies were pooled in statistical meta-analysis using Review Manager Software from the Cochrane Collaboration. Heterogeneity between combined studies was tested using standard chi-square test. Where statistical pooling was not appropriate or possible, the findings are summarised in narrative form. Results A comprehensive search of relevant databases, hand searching and cross referencing found 685 articles that were assessed for relevance to the review. Eighty-five papers appeared to meet the inclusion criteria based on title and abstract, and the full paper was retrieved. Of the 85 full papers reviewed, 40 were accepted for inclusion, three were systematic reviews, three were meta-analysis, and the remaining 34 were randomised controlled trials. For the randomised controlled trials that were able to be included in a meta-analysis, standardised mean differences or weighted mean differences and their 95% confidence intervals were calculated for each. Results from comparable groups of studies were pooled in statistical meta-analysis using Review Manager Software and heterogeneity between combined studies was assessed by using the chi-square test. Where statistical pooling was not appropriate or possible, the findings are summarised in narrative form. The results are discussed in two main sections. Firstly it was possible to assess the effectiveness of different types of caregiver interventions on the outcome categories of depression, health, subjective well-being, self-efficacy and burden. Secondly, results are reported by main outcome category. For each of these sections, meta-analysis was conducted where it was possible; otherwise, a narrative summary describes the findings. Effectiveness of intervention type Four categories of intervention were included in the review - psycho-educational, support, multi-component and other. Psycho-educational Thirteen studies used psycho-educational interventions, and all but one showed positive results across a range of outcomes. Eight studies were entered in a meta-analysis. No significant impact of psycho-educational interventions was found for the outcome categories of subjective well-being, self-efficacy or health. However, small but significant results were found for the categories of depression and burden. Support Seven studies discussed support only interventions and two of these showed significant results. These two studies were suitable for meta-analysis and demonstrated a small but significant improvement on caregiver burden. Multi component Twelve of the studies report multi-component interventions and 10 of these report significant outcomes across a broad range of outcome measures including self-efficacy, depression, subjective well-being and burden. Unfortunately because of the heterogeneity of study designs and outcome measures, no meta-analysis was possible. Other interventions Other interventions included the use of exercise or nutrition which resulted in improvements in psychological distress and health benefits. Case management and a computer aided support intervention provided mixed results. One cognitive behavioural therapy study reported a reduction in anxiety and positive impacts on patient behaviour. Effectiveness of interventions using specific outcome categories In addition to analysis by type of intervention it was possible to analyse results based on some outcome categories that were used across the studies. In particular the impact of interventions on caregiver depression was available for meta-analysis from eight studies. This indicated that multi-component and psycho-educational interventions showed a small but significant positive effect on caregiver depression. Five studies using the outcome category of caregiver burden were entered into a meta analysis and findings indicated that there were no significant effects of any of interventions. No meta-analysis was possible for the outcome categories of health, self-efficacy or subjective well-being. Implications for practice From this review there is evidence to support the use of well-designed psycho educational or multi-component interventions for caregivers of people with dementia who live in the community. Factors that appear to positively contribute to effective interventions are those which: * Provide opportunities within the intervention for the person with dementia as well as the caregiver to be involved * Encourage active participation in educational interventions for caregivers * Offer individualised programs rather than group sessions * Provide information on an ongoing basis, with specific information about services and coaching regarding their new role * Target the care recipient particularly by reduction in behaviours Factors which do not appear to have benefit in interventions are those which: * Simply refer caregivers to support groups * Only provide self help materials * Only offer peer support. PMID- 21631820 TI - The psychosocial spiritual experience of elderly individuals recovering from stroke: a systematic review. AB - Objectives The objective of this review was to appraise and synthesise best available evidence on the psychosocial spiritual experience of elderly individuals recovering from stroke. Inclusion criteria This review considered qualitative studies whose participants were adults, mean age of 65 years and older, and who had experienced a minimum of one stroke. Studies were included that described the participant's own experience of recovering from stroke. Search strategy The search strategy sought to find both published and unpublished studies and papers, not limited to the English language. An initial limited search of MEDLINE and CINAHL was undertaken followed by an analysis of text words contained in the title and abstract, and of index terms used to describe the article. A second extensive search was then undertaken using all identified key words and index terms. Methodological quality Each paper was assessed by two independent reviewers for methodological quality prior to inclusion in the review using the Qualitative Assessment and Review Instrument (QARI) developed by the Joanna Briggs Institute. Disagreements were resolved through consultation with a third reviewer. Data collection Information was extracted from each paper independently by two reviewers using the data extraction tool from QARI developed by the Joanna Briggs Institute. Disagreements were resolved through consultation with a third reviewer. Data synthesis Data synthesis aimed to portray an accurate interpretation and synthesis of concepts arising from the selected population's experience during their recovery from stroke. Results A total of 35 studies were identified and of those 27 studies were included in the review. These qualitative studies examined the perceptions of elderly individuals who had experienced a stroke. Findings were analysed using JBI-QARI. The process of meta synthesis using this program involved categorising findings and developing synthesised topics from the categories. Four syntheses were developed related to the perceptions and experiences of stroke survivors: sudden unexpected event, connectedness, reconstruction of life and life-altering event. Conclusion The onset and early period following a stroke is a confusing and terrifying experience. The period of recovery involves considerable psychological and physical work for elderly individuals to reconstruct their lives. For those with a spiritual tradition, connectedness to others and spiritual connection is important during recovery. The experience of stroke is a life-altering one for most elderly individuals, involving profound changes in functioning and sense of self. PMID- 21631821 TI - The role of patient isolation and compliance with isolation practices in the control of nosocomial MRSA in acute care. AB - Background Nosocomial infection remains the most common complication of hospitalisation. Despite infection control efforts, nosocomial methicillin resistant Staphylococcus aureus (MRSA) transmission continues to rise. Various isolation practices are used to minimise MRSA transmission in acute care. However, the effectiveness of these practices has seldom been evaluated. Objectives This review sought to evaluate the efficacy of isolation practices in minimising MRSA transmission in the acute hospital setting and explore staff, visitor and patient compliance with isolation practices. This review updates a review published in 2002. Search strategy A systematic search for relevant published or unpublished English language literature was undertaken using electronic databases, the reference lists of retrieved papers and the Internet. This extended the search published in the original review. Databases searched included: Medline, CINAHL, EMBASE, Cochrane Library and Joanna Briggs Institute Evidence Library. Selection criteria All English language research reports published between 1990 and August 2005 that focused on the role of isolation practices on the nosocomial transmission of MRSA in adult, paediatric or neonatal acute care settings were eligible for inclusion in the review. Studies that evaluated multiple infection control strategies or control of MRSA outbreaks were excluded. The main outcome of interest was the incidence of new cases of MRSA. The secondary outcome was staff, visitor and patient compliance with the isolation practices. Data collection and analysis Two reviewers assessed each paper against the inclusion criteria and a validated quality scale. Data extraction was undertaken using a tool designed specifically for this review. Statistical comparisons of findings were not possible, so findings are presented in a narrative form. Results Seven studies met the inclusion criteria. Given the small number of included studies and variable methodological quality, care must be taken when interpreting the review findings. There is some evidence that cessation of single room isolation and cohorting of MRSA patients does not increase nosocomial MRSA transmission when hand-washing compliance and standard precautions are maintained. Indeed, there is some evidence that reduced MRSA transmission can be achieved by improving compliance with contact precautions alone. The low level of hand hygiene compliance reported in the literature suggests that staff compliance with isolation practices is a significant factor in evaluating any infection-controlled intervention in the clinical setting. While staff compliance data are conflicting, regular audit and feedback of performance may improve compliance. Implications for clinical practice The heterogeneous nature of the topic and methodological weaknesses of included studies impairs the ability to aggregate data and develop specific practice recommendations. While this review presents evidence to suggest that ceasing single room or cohort isolation does not lead to increased MRSA transmission, these studies maintained high levels of hand hygiene or standard precautions. Additionally, the role of extraneous factors, such as environmental reservoirs, specific MRSA strains and patient mix, is unclear. None of the included studies measured financial, social or psychological factors associated with isolation practices. There is an urgent need for well-designed research with significant sample sizes to develop an evidence base upon which to underpin future clinical practice. PMID- 21631822 TI - A systematic review of psychometric evidence and expert opinion regarding the assessment of faecal incontinence in older community-dwelling adults. AB - Objectives This review had two objectives: (i) to determine what is required in an assessment of faecal incontinence issues for older community-dwelling adults; and (ii) to determine the psychometric tools most effective for assessment of faecal incontinence in older community-dwelling adults. Inclusion criteria For the review of psychometric tools, studies were included if they were concerned with people living in the community, included a significant proportion of the sample aged 65 years or over, and either examined psychometric properties of assessment tools or assessed sensitivity of assessment tools to non-surgical interventions available in the community setting. For the review of expert opinion, the search was limited to expert opinion provided by an expert in faecal incontinence that related to community-based assessment. Only articles published in English were eligible for inclusion and no limits were placed on publication dates. Search strategy An initial search of Medline and CINAHL databases identified terminology frequently used in the literature with regard to assessment of faecal incontinence. An extensive search was then undertaken using all identified key words and index terms. The third step involved a search of reference lists and bibliographies of all relevant articles. Methodological quality All identified studies that met the inclusion criteria were assessed for methodological validity in the case of studies considered for inclusion in the psychometric review. Validity of expert text was also assessed prior to it being included in the review. Results The final search strategy identified approximately 7000 references. Full-text versions of 144 references were critically appraised for inclusion in the review. Of these, 25 sources were included in the review of expert opinion and 16 in the review of psychometric properties. In the review of expert opinion, 254 conclusions were extracted for synthesis. The 31 thematic categories were organised under five major themes: History-taking, bowel assessment, psychosocial aspects, physical examination and specialist referral. From the sources that survived critical appraisal, 52 conclusions relating to psychometric properties of assessment tools were derived. There was limited, if any, analysis of psychometric properties for the majority of assessment tools. The Wexner and Vaizey symptom severity scales demonstrated acceptable test-retest reliability and convergent validity. The Fecal Incontinence Quality of Life Scale (FIQLS) demonstrated reasonable reliability and good convergent and criterion-related validity. There was, however, some evidence questioning its discriminant validity. Conclusions This systematic review represents an important first stage in developing guidelines for assessment of faecal incontinence in community-dwelling older people. Assessment should be comprehensive in nature. Gaps in expert opinion are evident regarding issues such as assessment of cognitive decline and specialist referral. Continence advisors need to be appropriately trained in using and interpreting results from assessment tools and conducting physical examinations. Although studies in the review of psychometric properties suffer from limitations such as inadequate sample sizes, the Vaizey and Wexner scales would appear to be the tools of choice. The FIQLS is clearly the tool of choice at this stage for measuring faecal incontinence quality of life. Further validation of tests used in faecal incontinence assessments is required. PMID- 21631823 TI - Improving and ensuring best practice continence management in residential aged care. AB - Background Continence Management within residential aged care is an every day component of care that requires assessment, implementation of strategies, resource allocation and evaluation. At times the management of incontinence of aged residents can be challenging and unsuccessful. The project chosen through the Clinical Fellowship program was Continence Management with the aim of raising awareness of best practice to assist in improving and providing person-centred resident care. Aims/objectives * Review the literature on best practice management of incontinence * Evaluate current practice in continence management for elderly residents within residential aged care services * Improve adherence to best practice strategies of care for incontinence * Raise awareness within the nursing home of the best practice management of incontinence * Promote appropriate and effective use of resources for continence management * Deliver individualised person-centred care to residents. * Ensure best practice in continence management Methods The Joanna Briggs Institute (JBI) Practical Application of Clinical Evidence System clinical audit tool was utilised to measure current practice against best practice. The results identify gaps that require improvement. The Getting Research into Practice process then allowed analysis of the level of compliance with each of the audit criteria, which would identify any barriers in implementing a selected course of action and aim to improve compliance. The project team was consulted with additional stakeholder consultation to form an action plan and implement strategies to improve practice. Results Although 100% compliance with all audit criteria in audit 1 and 2 was not achieved, there was improvement in the criteria concerning the documented fluid intake for residents. Further strategies have been identified and implemented and this continues to be a 'work in progress'. Staff now have an acute awareness of what best practice means and the impact their practices have on continence management. The JBI clinical audit and feedback cycle will continue to facilitate the measuring and implementation of best practice for resident outcomes in residential aged care. PMID- 21631824 TI - Implementation of best practice in advance care planning in an 'ageing in place' aged care facility. AB - Background For many older people, a residential aged care facility is home, a place where they should have the choice to die comfortably with a sense of control of care and treatment. An advance care plan describes the process of planning end-of-life care while a person is still able to make decisions with the aim of using this information should a person become unable to communicate their wishes. In the residential aged care setting, an advance care plan should be based on discussions involving the resident, family members and caregivers regarding the beliefs, values and goals of the resident and their family. It is not limited to the final days and weeks before dying, it can encompass a positive and open attitude towards death and dying. Aim The aim of this project was to examine the process of how residents' end-of-life care wishes is recorded and to ensure that the implementation of an advance care plan is performed according to the best available evidence. Method There were four stages to the project. The first stage involved interpretation of the five audit criteria. The criteria related to involving residents and their families in an advance care plan, providing them with appropriate information about end-of-life issues, and ensuring that staff are appropriately trained about an advance care plan. Next, the records of staff and resident were audited using JBI-PACES, the Joanna Briggs Institute's Practical Application of Clinical Evidence System. The next stage (Getting Research into Practice, GRIP) involved a process of situational analysis, action planning and action taking to improve compliance with best practice, followed by a post-implementation audit. Results Initially, compliance with an advance care plan best practice was fairly poor - less than 50% for each audit criterion. The GRIP phase highlighted seven barriers which were addressed during the implementation part of the project. These barriers included deficits related to the knowledge and education of residents, families and staff members, and issues related to administration and documentation, and concerns that any implementation process would not be sustainable. The post-implementation compliance was greater than 75% for each criterion. Conclusion The project highlights the ongoing challenges of educating residents and families, as well as staff on the importance of discussing/completing an advance care plan. Several improvements were made to the facility's documentation and records keeping processes, including the identification of resident's wishes regarding end-of life treatment. Residents and their families expressed a high level of satisfaction with these changes. Indeed, an increase in completed advance care plans at the facility had a clear benefit in improving care for residents towards the end of their life. PMID- 21631825 TI - In pursuit of clinical excellence: the role of evidence. PMID- 21631826 TI - Effectiveness of simulation on health profession students' knowledge, skills, confidence and satisfaction. AB - Background Despite the recent wave of interest being shown in high-fidelity simulators, they do not represent a new concept in healthcare education. Simulators have been a part of clinical education since the 1950s. The growth of patient simulation as a core educational tool has been driven by a number of factors. Declining inpatient populations, concerns for patient safety and advances in learning theory are forcing healthcare educators to look for alternatives to the traditional clinical encounter for skill acquisition for students. Objective The aim of this review was to identify the best available evidence on the effectiveness of using simulated learning experiences in pre licensure health profession education. Inclusion criteria Types of studies: This review considered any experimental or quasi-experimental studies that addressed the effectiveness of using simulated learning experiences in pre-licensure health profession practice. In the absence of randomised controlled trials, other research designs were considered for inclusion, such as, but not limited to: non randomised controlled trials and before-and-after studies. TYPES OF PARTICIPANTS: This review included participants who were pre-licensure practitioners in nursing, medicine, and rehabilitation therapy. Types of intervention(s)/phenomena of interest: Studies that evaluated the use of human physical anatomical models with or without computer support, including whole-body or part-body simulators were included. Types of outcome measures Student outcomes included knowledge acquisition, skill performance, learner satisfaction, critical thinking, self confidence and role identity. Search strategy Using a defined search and retrieval method, the following databases were accessed for the period 1995-2006: Medline, CINAHL, Embase, PsycINFO, HealthSTAR, Cochrane Database of Systematic Reviews and ERIC. Methodological quality Each paper was assessed by two independent reviewers for methodological quality prior to inclusion in the review using the standardised critical appraisal instruments for evidence of effectiveness, developed by the Joanna Briggs Institute. Disagreements were dealt with by consultations with a third reviewer. Data collection Information was extracted from each paper independently by two reviewers using the standardised data extraction tool from the Joanna Briggs Institute. Disagreements were dealt with by consultation with a third reviewer. Data synthesis Due to the type of designs and quality of available studies, it was not possible to pool quantitative research study results in statistical meta-analysis. As statistical pooling was not possible, the findings are presented in descriptive narrative form. Results Twenty-three studies were selected for inclusion in this review including partial task trainers and high-fidelity human patient simulators. The results indicate that there is high learner satisfaction with using simulators to learn clinical skills. The studies demonstrated that human patient simulators which are used for teaching higher level skills, such as airway management, and physiological concepts are useful. While there are short-term gains in knowledge and skill performance, it is evident that performance of skills over time after initial training decline. Conclusion At best, simulation can be used as an adjunct for clinical practice, not a replacement for everyday practice. Students enjoyed the sessions and using the models purportedly makes learning easier. However, it remains unclear whether the skills learned through a simulation experience transfer into real-world settings. More research is needed to evaluate whether the skills acquired with this teaching methodology transfer to the practice setting such as the impact of simulation training on team function. PMID- 21631827 TI - Fluid regimens for colostomy irrigation: a systematic review. AB - Background Various techniques for managing faecal evacuation have been proposed; however, colostomy irrigation is favoured as it leads to better patient outcomes. Alternative fluid regimens for colostomy irrigation have been suggested to achieve effective evacuation. Aim The objective of this review was to summarise the best available evidence on the most effective fluid regimen for colostomy irrigation. Search strategy Trials were identified by electronic searches of CINAHL, PubMed, MEDLINE, Current Contents, the Cochrane Library and EMBASE. Unpublished articles and references lists from included studies were also searched. Selection criteria Randomised controlled trials and before-and-after studies investigating any fluid regimen for colostomy irrigation were eligible for inclusion. Outcomes measured included fluid inflow time, total wash-out time, haemodynamic changes during irrigation, cramps, leakage episodes, quality of life and level of satisfaction. Data collection and analysis Trial selection, quality appraisal and data extraction were carried out independently by two reviewers. Differences in opinion were resolved by discussion. Main results The systematic literature search strategy identified two cross-over trials that compared water with another fluid regimen. Owing to the differences in irrigating solutions used, the results were not pooled for analysis. Both the polyethylene glycol electrolyte solution and glyceryl trinitrate performed significantly better than water. Conclusion There is some evidence to support the effectiveness of fluid regimens other than water, such as polyethylene glycol electrolyte and glyceryl trinitrate, for colostomy irrigation. Further well-designed clinical trials are required to establish solid evidence on the effectiveness of other irrigating solutions that might enhance colonic irrigation. PMID- 21631828 TI - The clinical effectiveness of Ginger (Zingiber officinale) in adults with osteoarthritis. AB - Background Osteoarthritis (OA) is a degenerative joint disease, which is associated with increased pain and disability, and a simultaneous decline in the quality of life of sufferers. While there is no cure for OA, there are numerous treatments that aim to reduce sufferers' symptoms and disability, and improve their quality of life. Medications, which have long been integral interventions for the management of OA, have recently been found to cause harm in some patients. Simultaneously, the increasing recognition of complementary and alternative therapies as part of mainstream health care, has seen many sufferers of OA use these therapies. Ginger has been commonly prescribed by herbalists for sufferers of OA due to its anti-inflammatory and circulatory stimulant effects. However, to date there has been no systematic review of the literature to evaluate the clinical effectiveness of Ginger for OA. Objective The objective of this systematic review was to evaluate the safety and effectiveness of Ginger in adults with OA. Data sources A comprehensive search was undertaken on 18 electronic databases from their inception to January 2007, including AARP Ageline, AMED, AMI, BioMed central gateway, CAM on PubMed, CENTRAL, CINAHL, Cochrane library, Current controlled trials, Current contents connect, DARE, Dissertations Abstract International, EMBASE, Health Source Nursing/Academic edition, International Pharmaceutical Abstract, MEDLINE, Natural medicines comprehensive database and TRIP. Review methods Randomised controlled trials or clinical controlled trials were sought, which evaluated the effectiveness of mono preparations of ginger in adults with OA of the knee or hip. Critical appraisal of study quality was undertaken using Joanna Briggs Institute critical appraisal instruments. Data extraction was via the Joanna Briggs Institute standard data extraction form for evidence of effectiveness. Results Five randomised controlled trials were identified from the search, of which three met the inclusion criteria. The methodological quality of the included studies was good. However, given that studies were clinically and methodologically heterogeneous, meta-analysis could not be conducted. Instead, evidence was summarised in narrative form. For changes in pain severity, studies comparing ginger extract (n = 110) to placebo (n = 111) reported mixed findings in support of the use of Ginger. Studies comparing ginger to an active control found participants who received Ibuprofen (n = 96) had a greater change in median pain intensity compared with participants who received Ginger (n = 110), and while findings were statistically significant for only one of the two studies, the results had limited clinical significance. Similarly, while two placebo-controlled studies reported differences between ginger (n = 70) and placebo (n = 71) for changes in disability and functional capacity, the difference was statistically and clinically significant for only one study. In one study comparing ginger to an active control, participants receiving Ibuprofen (n = 56) reported a statistically significant improvement in disability and functional capacity over time when compared with participants receiving Ginger (n = 56). In terms of safety, Ginger was well tolerated when compared with Ibuprofen, with infrequent reports of mild, and predominantly gastrointestinal, adverse effects. Conclusion Current evidence is weak for the use of Ginger in adults with OA of the knee and/or hip. Much of this can be attributed to significant heterogeneity between studies. Improvements in research design, instrumentation and ginger dosage, which more closely reflect current clinical practice, may help to demonstrate the safe and effective use of Ginger in OA sufferers. PMID- 21631829 TI - Understanding young peoples' experience of chronic illness: a systematic review. AB - Objective It has been reported that the diagnosis of a chronic illness increases a child's susceptibility to future mental health problems. Accordingly, the objective of the current review was to summarise the best available evidence that described a young person's experience of chronic illness and make recommendations towards the promotion of mental health and prevention of future mental health difficulties. Inclusion criteria The review considered qualitative research that used the voices of young people (under 18 years) to describe their experience of chronic illness and the impact it had on their lives. Search strategy The search strategy sought to find both published and unpublished research papers (limited to the English language). An extensive search was performed using the following databases: PubMed, CINAHL, Web of Science, PsycInfo, Aust Health, Dissertation Abstract International, Expanded Academic Index, Health Source Nursing, and Academic Search Elite. In addition, the reference lists of identified papers were hand searched, to capture all pertinent material, as well as relevant worldwide web sites. Methodological quality Each paper was assessed by two reviewers for methodological quality prior to inclusion in the review using the critical appraisal instrument (Qualitative Assessment and Review Instrument (QARI) ) from software developed by the Joanna Briggs Institute (JBI). Results A total of 18 qualitative papers were included in the review (nine grounded theory, six phenomenology, one ethnography, one social ecological, and one multiple case study). Forty-four papers were initially identified but 26 were excluded as they did not meet the inclusion criteria. Findings were extracted and meta-synthesised using JBI-QARI. Five syntheses about a young person's experience of chronic illness were derived: (i) the experience of chronic illness makes young people feel uncomfortable in their body and world; (ii) the experience of chronic illness disrupts 'normal' life; (iii) the experience of chronic illness is not all bad; (iv) ways of getting through the chronic illness experience, 'what others can do'; and (v) ways of getting through the chronic illness experience, 'what I can do'. Conclusion A positive perspective needs to be taken to promote mental health in young people with chronic illness; clinicians, families and interventions need to (i) bolster their sense of self; (ii) normalise the experience; (iii) foster its positive impact; (iv) help them accept the situation; and (v) help to develop the future-orientated coping strategies that will provide them with a sense of hope. PMID- 21631830 TI - Using Qualitative Assessment and Review Instrument software to synthesise studies on older people's views and experiences of falls prevention. AB - Aim To report on the experiences of using software (Qualitative Assessment and Review Instrument (QARI)) specifically designed to synthesise qualitative research studies. Background The synthesis of qualitative research using a systematic review process is somewhat contentious and at present a variety of approaches exists. One approach is the QARI tool from the Joanna Briggs Institute (JBI) which provides a structured process to qualitative synthesis for appraising, extracting and synthesizing qualitative research that is considered more aggregative than interpretative. The process shares similarities with the methods of quantitative systematic review although the analysis stage is grounded in qualitative methodology. QARI provides a means for accumulating and synthesizing qualitative-based knowledge on the view and experiences of healthcare users and staff that can be used alongside systematic reviews of effectiveness to supply information on barriers and facilitators to adopting an intervention. The synthesis of qualitative research on older people's experiences of falls is used as the focus in this paper. Conclusions In common with other qualitative synthesis methodologies, there remain important areas for debate. These relate to the process and outcomes of: quality assessment, extracting findings from primary studies, the creation of categories (themes, metaphors), the expression of the synthesis and the credibility of an iterative review process. PMID- 21631831 TI - Compliance with best practice: implementing the best available evidence in the use of physical restraint in residential aged care. AB - The Aged Care Clinical Fellowship, funded by the Commonwealth Department of Health and Ageing and conducted through the Joanna Briggs Institute is an initiative designed to improve the care of older Australians through clinical leadership and promotion of best practice. This paper outlines one of the projects undertaken at Carinya of Bicton, a residential aged high care facility, using an audit and feedback process to implement best practice standards in the use of physical restraint. Aims Between 12% and 47% of residents in residential care facilities are restrained; however, initial observation of residents restrained in the project facility showed that restraint devices were utilised in up to 40% of residents. Within the aged care sector there has been a shift in attitude to reducing or eliminating restraint in aged care facilities. Restraint is seen as a negative experience for the resident, being associated with physical discomfort, embarrassment and restriction of freedom and of movement. The purpose of the project was to improve practice in the area of physical restraint through the process of auditing current practice against evidence-based, best practice criteria and ultimately to reduce the level of restraint in the facility. Methods This practice improvement project utilised an audit and implementation cycle. The Joanna Briggs Institute Practical Application of Clinical Evidence System and best practice criteria developed from a systematic review were used to determine compliance with best practice. The Getting Research into Practice module was then employed to develop strategies to improve practice. Results The follow-up audit indicated there has been a reduction in the number of residents restrained, increased use of alternatives to restraint and an awareness on the part of all care staff of the policies and procedures, which govern the use of restraint in the facility. Conclusions It is recognised that the success of this project is in part due to the focus of all staff in the area and the support and assistance given to staff by management and the project team. This support will need to continue because while practice has improved in the short term, there are still barriers to change in this area. Also of benefit has been a shift in emphasis of the continuous quality improvement program at the facility to a more clinical focus. Management and staff, especially members of the project team have agreed that this process be utilised to improve practice in a number of other areas such as falls, constipation and behaviour management. PMID- 21631832 TI - Improving patient experiences of health care. PMID- 21631833 TI - The clinical effectiveness of length of bed rest for patients recovering from trans-femoral diagnostic cardiac catheterisation. AB - Background Cardiac catheterisation plays a vital role in the diagnosis and evaluation of cardiac conditions. The goal of management of patients after cardiac catheterisation is to reduce the risk of development of any local or prolonged vascular complications, in particular bleeding and haematoma formation at the puncture site. Bed rest and immobilisation of the affected leg are recommended practices to ensure adequate haemostasis at the femoral arterial puncture site and prevent complications. Objectives The objective of this review was to present the best available evidence for the optimal length of bed rest after trans-femoral diagnostic cardiac catheterisation. The main outcome of interest was the incidence of bleeding and haematoma formation following varying periods of bed rest. Search strategy We searched the following databases: CINAHL, Medline, Cochrane Library, Current Contents, EBSCO, Web of Science, Embase, British Nursing Index, Controlled clinical trials database, Google Scholar. Reference lists of relevant articles and conference proceedings were searched. We also contacted key organisations and researchers in the field. Selection criteria All randomised and quasi-randomised controlled trials that compared the effects of different lengths of bed rest following trans-femoral diagnostic cardiac catheterisation on patient outcomes were considered for inclusion in the review. Data collection and analysis Eligibility of the trials for inclusion in the review, details of eligible trials and the methodological quality of the trials were assessed independently by two reviewers. Odds ratios (OR) for dichotomous data and a weighted mean difference for continuous data were calculated with 95% confidence intervals (CI). Where synthesis was inappropriate, trials were considered separately. Main results Eighteen trials involving a total of 4294 participants were included in the review. One trial included three treatment groups. In seven trials among 747 people there was no significant difference in the incidence of bleeding following six or less than 6 h of bed rest (OR 1.47; 95% CI 0.60, 3.64). Likewise, there was no significant difference in the incidence of bleeding following bed rest at other time periods. In eight trials involving 2272 patients there was no significant difference in the incidence of haematoma formation following 6 or less than 6 h of bed rest (OR 0.82; 95% CI 0.59, 1.16). Significantly fewer patients randomised to less than 6 h of bed rest complained of back pain. The odds of developing back pain at 4 (OR 24.60; 95% CI 1.29, 469) and 24 h (OR 2.47; 95% CI 1.16, 5.23) following coronary catheterisation was significantly higher among patients randomised to 6 compared with 3 h of bed rest. Authors' conclusions There is evidence of no benefit relating to bleeding and haematoma formation in patients who have more than 3 h of bed rest following trans-femoral diagnostic cardiac catheterisation. However, there is evidence of benefit relating to decreased incidence and severity of back pain and cost-effectiveness following 3 h of bed rest. There is suggestive but inconclusive evidence of a benefit from bed rest for 2 h following trans-femoral cardiac catheterisation. Clinicians should consider a balance between avoiding increased risk of haematoma formation following 2-2.5 h of bed rest and circumventing back pain following more than 4 h of bed rest. PMID- 21631834 TI - Prophylactic steroids for paediatric open-heart surgery: a systematic review. AB - Background The immune response to cardiopulmonary bypass in infants and children can lead to a series of post-operative morbidities and mortality, that is, hemodynamic instability, increased infection and tachyarrhythmias. Administration of prophylactic doses of corticosteroids is sometimes used to try and ameliorate this pro-inflammatory response. However, the clinical benefits and harms of this type of intervention in the paediatric patient remain unclear. Objectives To systematically review the beneficial and harmful effects of the prophylactic administration of corticosteroids, compared with placebo, in paediatric open heart surgery. Search strategy The trials registry of the Cochrane Heart Group, the Cochrane Central Register of Controlled Trials in The Cochrane Library (Issue 4, 2006), MEDLINE (1966 to January 2007), EMBASE (1980 to January 2007) were searched. An additional hand-search of the EMRO database for Arabic literature was performed. Grey literature was searched, and experts in the field were contacted for any unpublished material. No language restrictions were applied. Selection criteria All randomised and quasi-randomised controlled trials of open heart surgery in the paediatric population that received corticosteroids pre-, peri- or post-operatively, with reported clinical outcomes in terms of morbidity and mortality. Data collection and analysis Eligible studies were abstracted and evaluated by two independent reviewers. All meta-analyses were completed using RevMan4.2.8. Weighted mean difference (WMD) was the primary summary statistic with data pooled using a random-effects model. Main results All cause mortality could not be assessed as the data reports were incomplete. There was weak evidence in favour of prophylactic corticosteroid administration for reducing intensive care unit stay, peak core temperature and duration of ventilation (WMD (95% confidence intervals) -0.50 h (-1.41 to 0.41); -0.20 degrees C (-1.16 to 0.77) and -0.63 h (-4.02 to 2.75) respectively). PMID- 21631835 TI - Systematic review of the effectiveness of nursing interventions in reducing or relieving post-operative pain. AB - Objectives The primary objective of this systematic review was to assess the effectiveness of nursing interventions for the relief/reduction of post-operative pain when compared with either standard care or other nursing interventions. Inclusion criteria Types of studies This review included randomised controlled trials (RCTs) and other quasi-randomised experimental research designs. Types of participants Adults treated for post-operative pain in an acute care hospital. Types of interventions Interventions included in the review were only those directly administered by nursing staff such as: (i) administration of analgesia: including oral, intramuscular injection, patient controlled analgesia, epidural; (ii) pre-operative patient education; (iii) assessment and documentation of intensity of pain at regular intervals; (iv) protocols, clinical pathways or flow sheets used in the management of pain; and (v) non-pharmacological interventions such as massage and relaxation. Types of outcome measures The primary outcome was the relief or reduction of post-operative pain. Other measures included analgesia consumption, patient satisfaction and length of hospital stay. Search strategy Search terms were chosen after reviewing text words and MeSH terms in relevant articles and databases. A search for published and unpublished research covering the period 1985-2004 (inclusive) was conducted, using 11 major electronic databases. Reference lists of retrieved articles were hand searched. The search was restricted to reports published in English. Assessment of quality The methodological quality of eligible studies was appraised by two independent reviewers, using a standardised critical appraisal tool. Differences in opinion were decided by consensus after discussion with a third investigator. Data extraction and analysis Data were extracted from the studies that met the criteria for methodological quality. Two reviewers independently extracted data for each study, using a standardised extraction tool. Results from homogeneous RCTs where possible were pooled in meta-analysis using RevMan4.2 software program. If the studies were clinically or statistically heterogeneous, the results are discussed in a narrative form. Non-randomised studies are also discussed in narrative form. Results Overall, there is no strong evidence to support the use of any intervention even though a few interventions showed some benefits. However, evidence for these benefits was often based on single studies. Most of the included studies showed that there was no difference between the interventions and the usual care with both being found equally effective. Implications for practice There was no strong evidence to support a particular practice. No intervention was found to be harmful; however, this does not presume to be evidence of safety. When there are two similarly effective interventions nurses need to weigh the possible positive and negative of the intervention including side effects, risk of adverse events, cost and patient preference. Other considerations include variations in patients' past pain experiences, type of surgery, many different analgesics. Although there were no controlled trials to support assessment and documentation, professionally and legally, documentation is required. PMID- 21631836 TI - Resistance exercises for health promotion in coronary patients: evidence of benefits and risks. AB - Aim This review aims to search and evaluate evidence related to benefits and risks of resistance exercises for patients with coronary artery disease (CAD), post myocardial infarction (MI) and its surgical treatment or coronary artery bypass grafting (CABG). Method An integrative literature review was carried out. The guiding question of this review was: What evidence is available regarding benefits and risks of muscle resistance training for patients with CAD, post-MI or CABG? Studies from January 1997 to August 2007 were included, taken from the following electronic databases: Medline, PEDro, Cochrane Controlled Trials Database and CINAHL. Results After the selection process, 11 articles were included in the review. Five articles presented just the resistance exercises, and six combined the resistance exercises with aerobic exercise. The benefits of intervention found in the articles were the improvement of peripheral muscular strength, increase of physical capacity, improvement of emotional state and quality of life. No risks were reported in the articles related to the application of the resistance exercise. Conclusions The resistance exercises can be used as a safe and effective strategy in health promotion programmes for coronary patients, making social, physical and psychological rehabilitation easier for those people. PMID- 21631837 TI - The infection control management of MRSA in acute care. AB - Background Many acute care facilities report endemic methicillin-resistant Staphylococcus aureus (MRSA), while others describe the occurrence of sporadic disease outbreaks. The timely implementation of effective infection control measures is essential to minimise the incidence of MRSA cases and the magnitude of disease outbreaks. Management strategies for the containment and control of MRSA currently vary between facilities and demonstrate varying levels of effectiveness. Objectives This review sought to systematically review the best available research regarding the efficacy of infection control practices in controlling endemic MRSA or MRSA outbreaks in the acute hospital setting. It updates an original review published in 2002. Search strategy A systematic search for relevant published or unpublished English language literature was undertaken using electronic databases, the reference lists of retrieved papers and the Internet. This extended the search of the original review. Databases searched included: Medline, CINAHL, EMBASE, Cochrane Library and Joanna Briggs Institute Evidence Library. Selection criteria All research reports published between 1990 and August 2005 in the English language that focused upon the infection control strategies that were implemented in response to either a nosocomial outbreak of MRSA or endemic MRSA within an acute clinical setting were included. Only studies that reported interventions which were implemented following the collection of baseline data were included. Data collection and analysis Two reviewers assessed each paper against the inclusion criteria and a validated quality scale. Data extraction was undertaken using a purposely designed tool. Given the heterogeneity of the interventions and outcomes measures, statistical comparisons of findings were not possible, therefore, the findings of this review are presented in a narrative format. Results Fourteen papers met the inclusion criteria for this review. Of these, 11 papers achieved a quality score above the threshold and were included in the review. Of the included papers, five describe MRSA outbreaks, while the remaining six describe endemic MRSA. All included studies used either exploratory descriptive or comparative designs. The varying combinations of interventions described in the included studies make it impossible to differentiate the efficacy of individual or even groups of interventions. The fact that most studies reported positive findings may reflect the bias towards publication of effective interventions. Implications for clinical practice This review provides evidence that the use of multifaceted strategies can reduce nosocomial MRSA transmission in both outbreaks of MRSA and settings where MRSA is endemic. The heterogeneous nature of the topic, combinations of interventions implemented and methodological weaknesses of the studies impairs the ability to aggregate data and develop specific recommendations for practice. PMID- 21631838 TI - Implementation of a glucose management protocol to prevent hypo- and hyperglycaemia in critically ill patients. AB - Background Many healthcare providers acknowledge the importance of encouraging health professionals to base their practice on current evidence. There is a plethora of evidence supporting tight glucose control in critically ill patients to reduce mortality and morbidity and many studies have shown that hyperglycaemia in critically ill patients has been associated with an increased susceptibility to infection and impaired immune response. Methods This project aimed to improve current practice in the prevention and management of hypo- and hyperglycaemia in critically ill patients based on the best available evidence. The setting for the project was a large public hospital in Brisbane, Queensland, with an 11 bed tertiary referral intensive care unit (ICU) for critically ill adult patients. A pre-implementation audit using a set of evidence-based criteria from a systematic review on tight glucose control in critically ill patients was undertaken. The Joanna Briggs Institute on-line audit and feedback software, paces (Practical Application of Clinical Evidence System), was used to analyse the data to determine ICU's current compliance with established best practice in glucose management. These initial audit results were used to develop an action plan to improve glucose management practice within the unit. This included the development and implementation of a glucose management protocol, the identification of barriers and facilitators and education of clinical staff. Following the implementation of this protocol a post-implementation audit was conducted using the same criteria as in the initial audit. Results The post audit findings indicated improvement in most areas of glucose management practice within the ICU. Conclusion The success of this project is not only evident through improved ICU glucose management practice over 12 weeks, but also through clinicians gaining experience in the process of utilising evidence-based recommendations in all areas of practice. PMID- 21631839 TI - The epistemology of patient safety research. AB - Patient safety has only recently been subjected to wide-spread systematic study. Healthcare differs from other high risk industries in being more diverse and multi-contextual, and less certain and regulated. Also many patient safety problems are low-frequency events associated with many, varied contributing factors. The subject of this paper is the epistemology of patient safety (the science of the method of finding out about patient safety). Patient safety research is considered here on the background of a risk management framework which requires researchers to: * Understand the context - as a subset of healthcare quality, services and systems research, with technical and human behavioural (cultural) components and a range of external and internal organisational influences, a wide range of research disciplines is necessary * Identify the risks - identify the things that go wrong and the frequency and nature of different types of incidents from sources such as medical record review, observational studies, audit, incident and medico-legal reports * Analyse the risks - deconstruct the things that go wrong, identifying contributing factors and trying to detect trends and patterns in contributing factors, detection, mitigation factors, ameliorating factors and actions taken to reduce risk * Evaluate the risks - decide on priorities, identifying preventive and corrective strategies and judging the risk- and cost-benefit of potential corrective strategies such as standardisation or simplification of a process or device * Manage the risk - evaluate and scope preventive and/or corrective strategies and then implement these, or place the problem on a risk register pending solution, or accept that what is needed is unaffordable * Communicate and consult - use interactive sessions, audit, on-going feedback, reminders and patient mediated prompts * Monitor and review the state of the problem - get baseline trends and patterns so that changes can be tracked and properly attributed to an intervention A hierarchy of levels of evidence has been proposed for clinical research and we argue that insufficient weighting has been given to lower ranked levels of research and to qualitative research, although critical interpretive synthesis is now gaining acceptance in mainstream thinking (e.g. by the Cochrane Collaboration). Fundamental challenges remain including how to grasp the elusive concept of patient safety, how to quantify, characterise and cost the problems, how to judge the extent to which harm can be attributed to errors, violations or system failures, how to identify contributing factors and the extent to which they can be implicated, how to judge whether incidents or their precursors are preventable, how to generate strong evidence to make healthcare safer and how to translate research into practice. Future directions include addressing the mundane as well as rare, dramatic events, and developing further research in non-hospital settings and in developing countries. In summary, a mixture of qualitative and quantitative methods, using information from all available data sources and combining retrospective, real time and prospective study designs, is necessary to address some of the more difficult patient safety problems. PMID- 21631841 TI - Elements of Evidence-based Healthcare. PMID- 21631842 TI - The clinical effectiveness of nurse practitioners' management of minor injuries in an adult emergency department: a systematic review. AB - Background The increasing cost of healthcare in Australia demands changes in the way healthcare is delivered. Nurse practitioners have been introduced into specialty areas including emergency departments. Specific interventions are known to include the treatment and management of minor injuries, but little has been reported on their work. Objectives Examine the best available evidence to determine the clinical effectiveness of emergency department nurse practitioners in the assessment, treatment and management of minor injuries in adults. Inclusion criteria For inclusion studies had to include adult patients treated for minor injuries by nurse practitioners in emergency departments. All study designs were included. Search strategy English language articles from 1986 onwards were sought using MEDLINE, CINAHL, Embase and Science Citation Index. Methodological quality Two independent reviewers critically appraised the quality of the studies and extracted data using standardised tools. Data collection Two independent reviewers assessed the eligibility of each study for inclusion into the review and the study design used. Where any disagreement occurred, consensus was reached by discussion with an independent researcher. Data synthesis Studies were assessed for homogeneity by considering populations, interventions and outcomes. Where heterogeneity was present, synthesis was undertaken in a narrative format; otherwise a meta-analysis was conducted. For each outcome measure, results were tabulated by intervention type and discussed in a narrative summary. Results from randomised controlled trials were pooled in meta-analyses where appropriate. Results Nine studies from a total of 55 participants met the inclusion criteria. Two were randomised controlled trials. Metasynthesis of research findings generated five synthesised findings derived from 16 study findings aggregated into seven categories. Evidence comparing the clinical effectiveness of nurse practitioners to mainstream management of minor injuries was fair to poor methodological quality. When comparable data were pooled, there were no significant differences (P < 0.05) between nurse practitioners and junior doctors. Conclusions The results emphasise the need for more high-quality research using appropriate outcome measures in the area of clinical effectiveness of nurse practitioners, particularly interventions that improve outcomes for presentations to emergency departments and address issues of waiting and congestion. PMID- 21631843 TI - The effectiveness of Cognitive-Behavioural Therapy with hopeful elements to prevent the development of depression in young people: a systematic review. AB - Background The onset of depression during adolescence can adversely impact future functioning. Cognitive-Behavioural Therapy (CBT) has been suggested to prevent depression in adolescence by providing an individual with the ability to interpret and the tools to deal with the impact of negative life events. Objective Examine the best available evidence to determine the effectiveness of CBT to prevent the onset of depression in young people, and assess whether the incorporation of hopeful elements makes CBT more effective. Search strategy A comprehensive three-step search strategy was developed to find both published and unpublished studies in English from 1987 to March 2007. Papers selected for retrieval were then assessed for methodological validity by two independent reviewers. Selection criteria Papers that used a randomised controlled design and investigated the efficacy of CBT to prevent the onset of depression in young people between the age of 10 years and 16 years were included. Papers were included if the CBT involved between four and 15 sessions, a follow-up period of between 3 and 24 months and included typical strategies, such as the identification of negative and irrational beliefs, the establishment of links between thoughts, feelings and behaviours, and provided tools so participants could self-monitor these. Data analysis Data were extracted using the standard tool from the Joanna Briggs Institute, pooled in a meta-analysis, and then grouped and analysed according to the amount of hopeful elements the CBT was judged to contain. Results and conclusion Limited evidence was found to indicate that CBT, regardless of its content (i.e. with or without hopeful elements), is effective at preventing the onset of clinical levels of depression in young people on a sustained basis. Nonetheless, given the devastating impact that depression can have on young people's future functioning, further research is needed to develop effective interventions to equip young people with the cognitive skills to buffer its onset on a more sustained basis and to enable them to reach and sustain mental health. PMID- 21631844 TI - The implementation and evaluation of an oral healthcare best practice guideline in a paediatric hospital. AB - The oral health of the New Zealand population now compares unfavourably with other countries. A number of strategies have been introduced at a government and health provider level to improve the oral health status of children. One such strategy was the introduction of a recommended best practice (RBP) within Starship Children's Hospital. Nursing practice was evaluated 2 weeks prior and 6 months post-implementation of the RBP using a survey technique. While there was no significant change in practice post-introduction of the RBP, awareness regarding the oral health care needs of children while in hospital has been improved. This study provides increased understanding in regard to the oral healthcare practices of paediatric nurses, the influences on evidence-based practice change and health education and promotion within an acute paediatric hospital. PMID- 21631845 TI - Effects of bariatric surgery on morbidity and mortality in severe obesity. AB - Background Obesity is associated with premature mortality, particularly when very severe and/or complicated by significant co-morbidities such as diabetes mellitus, cardiovascular and respiratory disease. Conventional management of obesity, namely diet, exercise, behavioural modification and pharmacotherapy has limited and poorly sustained effects on weight loss and uncertain benefits for survival. Objectives We aimed to review the literature in to determine whether bariatric surgery for morbid obesity prolongs life. Search Strategy A Search was conducted of data bases including Medline, Cochrane library, and Science Direct. Results Bariatric surgery produces significant and sustained weight loss. Greater weight loss occurs with procedures that have both a restrictive and malabsorptive component. In addition to resolution of, or at least improvement in co-morbidities and enhanced quality of life, six studies provide compelling evidence that bariatric surgery significantly prolongs life, an effect which is most marked in diabetics and predominantly attributable to reductions in death due to cardiovascular causes and cancer. Conclusion Taken together, the cost, quality of life, and survival benefits provide a compelling argument for the provision of bariatric surgery as a management strategy of choice for severe obesity, particularly when associated with diabetes mellitus, or other factors conferring a significant cardiovascular risk. The optimal procedure and strategy for patient selection remains to be determined. PMID- 21631846 TI - Information technology for medication administration: assessing bedside readiness among nurses in Lebanon. AB - Medication errors continue to be of great concern to hospitals. The use of Information technology (IT) for medication administration was recommended to assist nurses to administer medications safely, decrease the chance of medication errors, and contribute to patient safety. Such IT will be operational soon in some Lebanese hospitals. Users' readiness and acceptance to use such an IT application is crucial as it is a prerequisite for successful system implementation. This descriptive study used the Technology Acceptance Model to determine the level of nurses' readiness to use IT for medication administration in Lebanon. The sample included nurses working in three different major hospitals in Beirut. Data were collected on nurses' demographics, attitudes, perceived usefulness and ease of use of IT for medication administration. During the first 2 weeks of July, 2007, nurses manually or electronically were asked to voluntarily complete the questionnaire. Results showed that the users' attitude towards the use of the proposed IT is correlated with their perceptions on system usefulness and ease of use. Many showed a positive attitude towards system use and scored high on both perceptions. Yet around 20% of the nurses in the sample showed a negative attitude towards the use of the proposed system. PMID- 21631847 TI - Standardising care using integrated care pathways: if they seem like a solution, did we ask the right questions? PMID- 21631848 TI - Systematic review of the effectiveness of integrated care pathways: what works, for whom, in which circumstances? AB - Aim Integrated care pathways (ICP) are management technologies which formalise multidisciplinary team-working and enable professionals to examine their roles and responsibilities. ICPs are now being implemented across international healthcare arena, but evidence to support their use is equivocal. The aim of this study was to identify the circumstances in which ICPs are effective, for whom and in what contexts. Methods A systematic review of high-quality randomised controlled trials published between 1980 and 2008 (March) evaluating ICP use in child and adult populations in the full range of healthcare settings. RESULTS: 1 For relatively predictable trajectories of care ICPs can be effective in supporting proactive care management and ensuring that patients receive relevant clinical interventions and/or assessments in a timely manner. This can lead to improvements in service quality and service efficiency without adverse consequences for patients. 2 ICPs are an effective mechanism for promoting adherence to guidelines or treatment protocols thereby reducing variation in practice. 3 ICPs can be effective in improving documentation of treatment goals, documentation of communication with patients, carers and health professionals. 4 ICPs can be effective in improving physician agreement about treatment options. 5 ICPs can be effective in supporting decision-making when they incorporate a decision-aide. 6 The evidence considered in this review indicates that ICPs may be particularly effective in changing professional behaviours in the desired direction, where there is scope for improvement or where roles are new. 7 Even in contexts in which health professionals are already experienced with a particular pathway, ICP use brings additional beneficial effects in directing professional practice in the desired direction. 8 ICPs may be less effective in bringing about service quality and efficiency gains in variable patient trajectories. 9 ICPs may be less effective in bringing about quality improvements in circumstances in which services are already based on best evidence and multidisciplinary working is well established. 10 Depending on their purpose, the benefits of ICPs may be greater for certain patient subgroups than others. 11 We do not know whether the costs of ICP development and implementation are justified by any of their reported benefits. 12 ICPs may need supporting mechanisms to underpin their implementation and ensure their adoption in practice, particularly in circumstances in which ICP use is a significant change in organisational culture. 13 ICP documentation can introduce scope for new kinds of error. Conclusions ICPs are most effective in contexts where patient care trajectories are predictable. Their value in settings in which recovery pathways are more variable is less clear. ICPs are most effective in bringing about behavioural changes where there are identified deficiencies in services; their value in contexts where inter-professional working is well established is less certain. None of the studies reviewed included an economic evaluation and thus it is not known whether their benefits justify the costs of their implementation. PMID- 21631849 TI - Policy related to abdominoplasty in publicly funded elective surgery programs: a systematic review. AB - Objectives This systematic review set out to establish best practice in relation to policy for the inclusion/exclusion of abdominoplasty procedures within public health systems. Inclusion criteria The review considered any studies relating to abdominoplasty that addressed issues of inclusion/exclusion from public funded health systems including criteria for clinical need, contraindications, fit/ready for surgery, policy compliance and issues in relation to surgical training. Search strategy The search strategy sought to find published and unpublished studies and papers limited to English. An initial search of Medline and CINAHL was undertaken, followed by an analysis of keywords contained in the title, abstract and index terms. A second comprehensive search was then undertaken using Medline, CINAHL, EMBASE, AUSTROM, Health Business, and FullTEXT Elite and PsycINFO. The search was restricted to the period 1995-2005. Methodological quality Each paper identified was assessed by two independent reviewers for methodological quality before inclusion in the review using an appropriate critical appraisal instrument from the Joanna Briggs Institute System for the Unified Management and Assessment Review Instrument package. Results A total of 19 papers were included in the review. Owing to the diverse nature of the papers no meta-analysis or meta-synthesis was able to be used to pool studies. The results are therefore presented in a narrative form. The papers identified were mainly retrospective audits and discussion/opinion papers. The main issues addressed were criteria to establish clinical need, contraindications and policy compliance. Conclusion There are clinical indicators, mainly in relation to physical symptoms/dysfunction, to support exemption of some cases of abdominoplasty. For abdominoplasty to be conducted clinical need must be assessed and formally documented. Where clinical need is primarily based on psychological distress/dysfunction a formal psychiatric assessment should be used to justify surgery. PMID- 21631850 TI - Policy related to rhinoplasty in publicly funded elective surgery programs: a systematic review. AB - Objectives This systematic review set out to establish best practice in relation to policy for the inclusion/exclusion of rhinoplasty within public health systems. Inclusion criteria The review considered any studies relating to rhinoplasty that addressed issues of inclusion/exclusion from public funded health systems including criteria for clinical need, contraindications, fit/ready for surgery, policy compliance and issues in relation to surgical training. Search strategy The search strategy sought to find published and unpublished studies and papers limited to English. An initial search of Medline and CINAHL was undertaken, followed by an analysis of keywords contained in the title, abstract and index terms. A second comprehensive search was then undertaken using Medline, CINAHL, EMBASE, AUSTROM, Health Business, and FullTEXT Elite and PsycINFO. The search was restricted to the period 1995-2005. Methodological quality Each paper was assessed by two independent reviewers for methodological quality before inclusion in the review using an appropriate critical appraisal instrument from the Joanna Briggs Institute System for the Unified Management and Assessment Review Instrument package. Results A total of nine papers were included in the review. Owing to the diverse nature of these papers, meta synthesis was only possible for three of the seven papers. For this reason, the rest of the results are presented in narrative form. The majority of the papers included in the review related to psychological indicators for surgery and the potential impact of surgery on quality of life. Conclusion Where clinical indications for surgery are determined and documented following thorough diagnostic evaluation, rhinoplasty may be conducted. PMID- 21631851 TI - Evidence-based review of therapies at the menopause. AB - Background and Objective The highest level of scientific evidence available for each therapy for menopausal symptoms was sought, for example, systematic reviews of randomised controlled trials (RCTs). Results There is reasonable evidence that some symptoms are modified by lifestyle, for example, cessation of smoking, exercise, reduction of alcohol, diet and alleviation of psychosocial stress. No complementary medicine, for example, phytoestrogens, black cohosh, herbal or homeopathic medicines or complementary therapies, for example, acupuncture, yoga, chiropractic manipulation, reflexology or magnetic devices have a greater effect than the usual placebo effect seen in quality blinded RCTs. Some have potential side-effects. So-called 'bioidentical hormones' have no evidence-base and potential for harm. None of the above therapies have evidence of efficacy and long-term safety. Selective serotonin and noradrenaline re-uptake inhibitors ameliorate vasomotor symptoms and sometimes menopausal depression better than placebo. The most effective therapy for menopausal (oestrogen) deficiency symptoms is oestrogen which is the main component of hormone replacement therapies (HRT). Compared with placebo HRT is highly effective in relieving hot flushes, night sweats, dry vagina and dyspareunia. It also improved joint pains, sexuality and sleeplessness and reduced subsequent fractures in RCTs. The increased risk of oral HRT for thromboembolism is small around menopause, for those without thrombotic risk factors, and is not elevated with non-oral routes. Cardiovascular disease may be reduced when HRT is initiated near menopause. Breast cancer risk increases after several years with the use of oral HRT containing progestogens at an annual rate of 8/10 000 (<0.1%). No increase in breast cancer risk was seen with oestrogen-only HRT. PMID- 21631852 TI - Knowledge translation and guidelines: a transfer, translation or transformation process? AB - Aim The case is made for conceptualising guidelines as more flexible, interpretative pieces of information that contain a mixture of factual evidence, evidence that requires interpretation and evidence that will be shaped and moulded by the particular context into which it will be introduced. By acknowledging this way of viewing guidelines, practitioners and researchers are more able to understand what works in terms of successful implementation. This perspective differs from rating the type of evidence within guidelines according to the nature of the evidence. Methods The approach used was based on a standard process of formulating theoretical arguments: presenting the background, making and justifying the case by presenting evidence on successful guideline implementation strategies. Having presented the reasons why we should relook at the way we conceptualise and use guidelines, the paper then contrasts the evidence derived from research with examples of practitioners' accounts of managing the introduction of guidelines in practice. From this a number of conceptualisations are presented that offer new ways of constructing multifaceted interventions for guideline implementation. Results A set of arguments has been presented that provide background evidence, draw broad conclusions, address the ongoing issues around non-spread of new information and offer different ways of constructing research studies. The arguments are built on empirical and experiential evidence and refer to several major studies. Conclusions Seven conclusions are drawn from the paper: guidelines are not 'literal' objects - they are complex communication vehicles that are used as catalysts to stimulate discussion, learning and debate across knowledge boundaries; current evidence on guideline implementation reinforces the need to look at complex, multifaceted interventions based on specific diagnosis of barriers to implementation; this process poses conceptual, theoretical and methodological challenges to the research community; research designs ought to consider the contribution of process models as well as causal models; the implication of such a move would be investment in more process studies before the setting up of expensive causal or intervention (RCT type) studies; refocusing of implementation research away from theories of behaviour change to more consideration of knowledge management is to be encouraged and organisational theory and theory formulation and testing ought to be heterogeneous rather than narrowly focused. PMID- 21631853 TI - The implementation of evidence-based practice for the prevention/management of post-operative nausea and vomiting. AB - Aim To bring the research evidence for the prevention/management of post operative nausea and vomiting into clinical practice in the surgical services department of a community hospital. Methods Audit and feedback were used to lead organisational change at the project hospital. A team of key stakeholders was created and helped to bring change to the way post-operative nausea and vomiting was treated and managed at the institution. The Chief of Anesthesia, Assistant Chief Nursing Officer, Nurse Researcher, Director of Surgical Services, operating room manager, post-anaesthesia care unit manager, pre-admission testing nurse and author as team leader made up the change management team. Current compliance with the six criteria set forth as research evidence for the change was derived from a surgical chart audit. Strategies for changes to increase compliance were then developed by the team. For the next several months change management occurred. Results There was a vast improvement in evidence-based practice following change management. Furthermore, post-operative nausea and vomiting decreased from 18% at the start to 5% nausea and 0% vomiting at the end. Conclusion Change management using audit and feedback is effective in changing organisational practice and in improving patient outcomes following surgery. PMID- 21631854 TI - Literature review to identify factors that support implementation of evidence based practice in residential aged care. AB - The aim was to undertake a review of the literature on change management, quality improvement, evidence-based practice and diffusion of innovations to identify key factors that might influence the uptake and continued use of evidence in residential aged care. The key factors will be used to shape and inform the evaluation of the Encouraging Best Practice in Residential Aged Care Program which commenced in Australia in 2007. MEDLINE, CINAHL and the Cochrane Database of Systematic Reviews were searched using combinations of search terms. Searching focused on existing literature reviews, discussions of relevant conceptual and theoretical frameworks and primary studies that have examined the implementation of evidence-based practice in residential aged care. Keyword searching was supplemented with snowball searching (following up on the references cited in the papers identified by the search), searching by key authors in the field and hand searching of a small number of journals. In general, the period covered by the searches was from 2002 to 2008. The findings from the literature are often equivocal. Analysis and consolidation of factors derived from the literature that might influence the implementation of evidence-based practice resulted in the identification of eight factors: (i) a receptive context for change; (ii) having a model of change to guide implementation; (iii) adequate resources; (iv) staff with the necessary skills; (v) stakeholder engagement, participation and commitment; (vi) the nature of the change in practice; (vii) systems in place to support the use of evidence; and (viii) demonstrable benefits of the change. Most of the literature included in the review is from studies in healthcare and hence the generalisability to residential aged care is largely unknown. However, the focus of this research is on clinical care, within the context of residential aged care, hence the healthcare literature is relevant. The factors are relatively broad and cover the evidence itself, the process of implementation, the context within which evidence will be implemented and the systems and resources to support implementation. It is likely that the factors are not independent of each other. The set of factors will be refined over the course of the evaluation. PMID- 21631855 TI - Implementation of evidence. PMID- 21631856 TI - Heart failure self-management education: a systematic review of the evidence. AB - OBJECTIVE: The objective of this systematic review is to identify educational content and techniques that lead to successful patient self-management and improved outcomes in congestive heart failure education programs. METHODS: MEDLINE, CINAHL and the Cochrane Central Register of Controlled Trials, as well as reference lists of included studies and relevant reviews, were searched. Eligible studies were randomised controlled trials evaluating congestive heart failure self-management education programs with outcome measures. Two of the investigators independently abstracted descriptive information, education content topics and outcomes data. RESULTS: A total of 7413 patients participated in the 35 eligible congestive heart failure self-management education studies. The congestive heart failure self-management programs incorporated 20 education topics in four categories: (i) knowledge and self-management (diagnosis and prognosis, pathophysiology of how congestive heart failure affects the body, aims of treatment, management and symptoms, medication review and discussion of side effects, knowing when to access/call the general practitioner, communication with the physician, follow up for assessment or reinforcement); (ii) social interaction and support (social interaction and support, stress, depression); (iii) fluids management (sodium restriction, fluid balance, daily measurement of weight, ankle circumference, self-monitoring and compliance relative to fluids); and (iv) diet and activity (dietary assessment and instructions, physical activity and exercise, alcohol intake, smoking cessation). A total of 113 unique outcomes in nine categories (satisfaction, learning, behaviour, medications, clinical status, social functioning, mortality, medical resource utilisation and cost) were measured in the studies. Sixty (53%) of the outcomes showed significant improvement in at least one study. CONCLUSION: Educational interventions should be based on scientifically sound research evidence. The education topic list developed in this review can be used by patients and clinicians to prioritise and personalise education. PMID- 21631857 TI - Use of qualitative research as evidence in the clinical guideline program of the National Institute for Health and Clinical Excellence. AB - AIM: To describe the use of qualitative research as evidence in a national clinical guideline program (National Institute for Health and Clinical Excellence - NICE, UK) and to identify training needs for guideline developers. METHODS: All published NICE clinical guidelines from December 2002 to June 2007 were reviewed to determine whether qualitative studies were considered as evidence in the development of recommendations and how this type of evidence had been used. Developers of clinical guidelines due to be published between July 2007 and March 2008 were asked to describe their training needs regarding the use of qualitative research in clinical guidelines. Data were summarised using simple descriptive statistics. RESULTS: Of the 49 clinical guidelines published by NICE within the study period, nearly half (45%, 22/49) used qualitative studies as an evidence base for developing recommendations for clinical practice. The number of qualitative studies used in these clinical guidelines increased from 2003 to 2006: 9 studies in 2003; 41 studies in 2004; 60 studies in 2005; 139 studies in 2006. In terms of how qualitative evidence was used in the guidelines, the study identified the following main issues: inconsistencies in the terminology used to describe types of qualitative study design; lack of standardised search strategies and/or targeted processes to select studies from databases; lack of a standardised approach to quality appraisal and poor reporting of how the identified evidence was used to inform the relevant guideline recommendations. Of the 17 clinical guidelines in development during the study period, the questionnaire was returned by approximately half of the guideline developers (response rate 47%, 8/17). A wide range of training needs was identified, chiefly training in the identification, quality appraisal and synthesis of qualitative studies and guidance as to the guideline areas where qualitative studies should be considered as evidence. CONCLUSION: Qualitative research is increasingly being used by NICE's clinical guideline developers as an evidence base to generate clinical practice recommendations. There are, however, clear training needs for NICE's guideline developers in terms of how best to identify, quality appraise and synthesise qualitative evidence for use in evidence-based clinical guidelines. PMID- 21631858 TI - Are we providing the best possible pain management for our elderly patients in the acute-care setting? AB - BACKGROUND: During 2008 seven practice improvement projects were conducted in an acute-care hospital under the banner of The Older Person and Improving Care (TOPIC 7). Each project team examined a discrete aspect of care of the elderly and this project focus was on pain management and in particular assessment of the older person with communication difficulties. AIMS: The project intended to assess current practice and implement changes to match best practice in the management of pain in the older person within an acute-care setting. METHODS: A multidisciplinary team was recruited to conduct the project. The pain team with the other six TOPIC 7 teams was facilitated by a coordinating team. The project was divided into four phases. Phase one was designated as Describing, where the clinical issue was identified and focused to priority areas. It was decided to focus on the elderly who were unable to verbally communicate their pain management needs. Standards of practice relevant to the area of practice were sourced to guide practice improvement. Phase two was Measuring activity where clinical audits were used to measure current practice and compare this with the appropriate standards. Phase three was Taking action where a range of practice improvement activities were implemented including the introduction of the Abbey Pain Scale. The final phase was designated as Review and share where the impact of the project activities was measured with a follow-up audit and the results were disseminated. RESULTS: Initial results showed a need for increased awareness of the difficulties in the pain assessment and management of older persons who cannot verbally communicate their needs. Seventy-eight per cent of patients had a documented pain assessment in the previous 24 h on audit. However, 83% of these assessments were only a general comment in the patient records. No tools were available for completing a systematic assessment that was reliable and reproducible. Sixty-two per cent of patients did not have analgesia administered 1 h before mobilising or having a significant dressing completed. Reaudit, posteducation sessions, showed an increase in the awareness in the complexities involved in caring for this group of patients. An alternate, specific tool (Abbey Pain Scale) was utilised well by staff and acted as a prompt in reminding staff to assess the pain levels of their elderly patients. CONCLUSIONS: Globally, pain in the older person is poorly managed. Pain in older persons who are unable to communicate or who have difficulty in communicating their needs is even more poorly managed. However, the availability of an appropriate tool and an increased awareness of this issue can have a significant and real impact on the pain management of this group of patients. PMID- 21631859 TI - Reducing functional decline of older people in an acute-care setting: are we providing adequate care to maintain/optimise the functional status of our elder patients? AB - AIMS: To improve practice in the care of the older person to ensure their functional status has not declined at the point of discharge by: determining available standards of care in relation to assessment of functional ability in the elderly; measuring current practice against standards of care based on the best available evidence; establishing priority areas for practice/improvement; implementing the practice/improvement plan; measuring practice to determine improvement in the management of functional decline in older patients and identifying further areas for investigation. METHODS: A multidisciplinary team used a range of knowledge translation tools and strategies in order to reduce functional decline of the older person in the acute-care setting. Audit was used to determine compliance with best practice standards and to determine the level of functional decline. A novel intervention was introduced in the form of a communal dining room and this was evaluated by the project team. The project was part of a program of seven projects (The Older Person and Improving Care) facilitated by a team with expertise in knowledge translation. Facilitation was provided though a structured education program and ongoing support. RESULTS: It was identified for this organisation that it was necessary to develop specific standards of care for the elderly in relation to functional decline. It was confirmed that there were unacceptable levels of functional decline in the elderly particularly in relation to mobility. The project team were able to show the feasibility and potential benefits of a communal dining room in reducing functional decline. CONCLUSIONS: It has been recognised that practice improvement activities are complex and that despite good planning and facilitation things do not always go to plan. This stated, it should be recognised that a great deal was achieved. Standards of care were able to be developed from an evidence base. These standards were used to develop an audit tool that confirmed deficits in practice. The outcome was a trial of a new intervention, a communal dining room, which showed considerable potential in reducing functional decline in the elderly. PMID- 21631860 TI - Benefits of implementing an interdisciplinary and multifactorial strategy to falls prevention in a rural, residential aged-care facility. AB - BACKGROUND: Falls are a frequent and serious problem facing residents in aged care facilities. Their prevention continues to be one of the most challenging aspects of the increasingly complex health-care needs of the aged. A fall can impact hugely on a resident's quality of life, health and health-care costs. Evidence supports the completion of a falls risk assessment and a program of multiple interventions as successful measures in reducing falls. OBJECTIVE: The aim of this project was to improve the local practice in the prevention of falls and to ensure this practice is performed according to the best available evidence. METHOD: The Joanna Briggs Institute Practical Application of Clinical Evidence System program was the clinical audit tool used. This program utilised a process of audit, feedback and reaudit as a strategy to improve practice. This was to be completed by developing a strategy to implement an interdisciplinary and multifactorial strategy to falls prevention in a rural, residential aged-care facility. A program of multiple interventions that were aimed at minimising individual client's risk of falling was utilised in this project. RESULTS: The 20 week intervention program of this project had a positive effect on audit criterion that supports the evidence-based practice for an interdisciplinary, multifactorial falls prevention program. The number of residents falling and the number of falls occurring were significantly reduced. CONCLUSION: An interdisciplinary and multifactorial prevention program may reduce falls in an aged-care facility. The Joanna Briggs Institute Practical Application of Clinical Evidence System program proved to be an effective and easy-to-use tool to stimulate clinical change in the workplace. Its cycle of audit, feedback and reaudit can easily be utilised to improve other clinical issues relevant to our facility. PMID- 21631861 TI - Using all the available evidence: perceptions of paediatric occupational therapists about how to increase evidence-based practice. AB - AIM: This study aimed to (i) explore occupational therapy practitioners' perceptions of the scope and sources of evidence that they use in clinical decision-making; and (ii) identify strategies and resources that would increase the use of evidence in a paediatric clinical service. METHODS: An action research methodology was applied to systematically explore participants' experiences of using evidence in practice and to democratically develop ways of increasing evidence-based practice. A series of six focus groups were conducted with nine occupational therapists, whose clinical experience ranged from 5 to 24 years. All participants had completed or were undertaking research qualifications. A systematic process of data coding was used whereby data were coded, categorised, analysed thematically and summaries developed. Rigour was maintained through triangulation (i.e. multiple data sources and occasions of data collection), and consultation and interpretation by two researchers. RESULTS: Participants identified two distinct sources of evidence they use in clinical practice: research-based and practice-based. They identified features of each source that they valued but also described barriers to the utilisation of each. Participants proposed strategies to increase the value of each source of evidence in their clinical practice. Strategies included appraising the clinical relevance and utility of research-based evidence, and developing standards and systematic processes to increase the credibility of practice-based evidence. CONCLUSIONS: Participants in this study perceived that although it is necessary to use all the available evidence in practice settings, there are currently barriers to using both research-based and practice-based evidence. In particular, the study indicates that there is need for the development of a structured model with processes and standards to guide practitioners in the collection, trial, evaluation and dissemination of practice-generated evidence. Further research is needed to explore the extent to which valid evidence can be generated from individual client programs by examining therapists' strategies for goal attainment in practice settings. PMID- 21631863 TI - Nutritional frailty: unrecognised it threatens independence. PMID- 21631864 TI - Review of systematic reviews about the efficacy of non-pharmacological interventions to improve sleep quality in insomnia. AB - Background Insomnia is a very common condition in various populations. Non pharmacological interventions might offer (safe) alternatives for hypnotics. Aim To evaluate the evidence for efficacy from systematic reviews about non pharmacological interventions to improve sleep quality in insomnia by a systematic review of systematic reviews and meta-analyses. Search strategy Search strategies were conducted in the Database of Abstracts of Reviews of Effects (2002-July 2008), The Cochrane Database of Systematic Reviews (2000-July 2008) and PubMed (1950-July 2008). Sleep quality was the outcome measure of interest. Selection criteria Systematic reviews about the efficacy of one or more non-pharmacological interventions for insomnia, concerning both adult and elderly populations, were included. Reviews that included studies performed among populations suffering with severe neurological or cognitive impairments or with addictive disorders were excluded. Data analysis Relevant data were extracted. The quality of the reviews found was appraised by using the Overview Quality Assessment Questionnaire. The evidence was appraised and divided into six classes. Results and conclusions Sixteen reviews about 17 interventions were included. Six reviews were of adequate methodological quality. Of these, only one provided an effect size: a moderate effect was found for music-assisted relaxation. Weak evidence indicating a large effect was found for multicomponent cognitive behavioural therapy, progressive muscle relaxation, stimulus control and 'behavioural only'. Weak evidence indicating a moderate effect was found for paradoxical intention. Finally, weak evidence indicating a moderate to large effect was found for relaxation training. Because of the lack of sufficient methodological quality and the lack of calculated effect sizes, most of the included reviews were not suitable for drawing rigorous conclusions about the effect of non-pharmacological interventions on sleep quality in insomniacs. The non-pharmacological treatment of insomnia would benefit from renewed reviews based on a rigorous methodological approach. PMID- 21631865 TI - Interventions to reduce the incidence of falls in older adult patients in acute care hospitals: a systematic review. AB - Aim Falls can cause serious physical and emotional injuries to patients leading to poor quality of life and increased length of hospital stay. The aim of this study was to present the best available evidence regarding the effectiveness of risk assessment or other interventions that aimed to minimise the number of falls. Methods A systematic review of randomised controlled trials was undertaken to determine the effectiveness of interventions that were designed to reduce the incidence of falls in older acute-care patients. Only trials published between 1998 and 2008 were considered. Results Only seven studies were included in the review, indicating the evidence on this topic is sparse. There is some evidence to suggest that implementing the following interventions in acute hospitals may be effective in reducing the amount of falls of older adult inpatients: * A multidisciplinary multifactorial intervention program consisting of falls risk alert card, an exercise program, an education program and the use of hip protectors after approximately 45 days * A one-on-one patient education package entailing information on risk factors and preventative strategies for falls as well as goal setting * A targeted fall risk factor reduction intervention that includes a fall risk factor screen, recommended interventions encompassing local advice and a summary of the evidence There is also some evidence to suggest that implementing a multidisciplinary multifactorial intervention that consists of systematic assessment and treatment of fall risk factors, as well as active management of postoperative complications, can reduce the amount of falls in patients following surgery for femoral neck fracture. Conclusion There is some evidence to suggest that certain multifactorial interventions are more effective than others and that increasing patient education or targeting fall risk factors may be of benefit. Further high-quality research is needed in order to ascertain effective fall-prevention strategies in acute-care facilities. PMID- 21631866 TI - Policy related to varicose veins in publicly funded elective surgery programs: a comprehensive systematic review. AB - Objectives This systematic review set out to establish best practice in relation to policy for the inclusion/exclusion of varicose vein procedures within public health systems. Inclusion criteria The review considered any studies relating to varicose veins that addressed issues of inclusion/exclusion from publicly funded health systems including criteria for clinical need, contraindications, fit/ready for surgery, policy compliance and issues in relation to surgical training. Search strategy The search strategy sought to find published and unpublished studies that were limited to English. An initial search of Medline and CINAHL was undertaken, followed by an analysis of key words contained in the title, abstract and index terms. A second comprehensive search was then undertaken using Medline, CINAHL, EMBASE, AUSTROM, Health Business FullTEXT Elite and PsycINFO. The search was restricted to the period 1995-2005. Methodological quality Each paper was assessed by two independent reviewers for methodological quality before inclusion in the review using an appropriate critical appraisal instrument from the Joanna Briggs Institute System for the Unified Management, Assessment and Review of Information software (JBI-SUMARI). Results A total of nine papers were included in the review. Because of the diverse nature of these papers, meta-analysis and synthesis of the results was not possible. For this reason, results are presented in narrative form. The majority of papers included in the review related to the identification of indicators for surgical intervention for the treatment of varicose veins. Other major themes were related to measures of quality of life of patients with varicose veins and the potential for surgical training. Recommendations for practice and research 1 Reasons for presentation should be thoroughly examined before any decision is made regarding surgery for varicose veins and indications documented thoroughly. 2 Every effort should be made to ensure opportunities are available to trainees within the constraints of the system. 3 In considering varicose veins surgery specifically, there would be value in examining further the use of psychometric scales/instruments to assess individual need for surgery. 4 The financial impact of varicose veins surgery should be investigated by way of a cost utility study. 5 Further investigation of the impact of excluding varicose vein surgery on training would be warranted. PMID- 21631867 TI - Physical leisure activities and their role in preventing dementia: a systematic review. AB - Aim To synthesise the best available evidence concerning the role of physical leisure activities in preventing dementia among older adults. Methods Studies containing adults aged 60 years and older with or without a clinical diagnosis of dementia who did or did not participate in physical leisure activities were considered. Activities were those that required active movement of the body such as gardening or playing sports, which were not for occupation-related purposes nor activities of daily living. Experimental and observational studies in the English language were targeted; there was no date restriction. Results A total of 17 epidemiological studies were included in the review. The evidence was equivocal regarding the relationship between participation in physical activities during midlife and later life and the prevention of dementia. Conclusion Participating in physical activities during middle and later adult life can be neither refuted nor recommended to prevent the onset of dementia. Engaging in some physical activities (i.e. gardening, walking) appears to be more beneficial than engaging in other activities. PMID- 21631868 TI - Improving the fundamentals of care for older people in the acute hospital setting: facilitating practice improvement using a Knowledge Translation Toolkit. AB - This paper reports on a structured facilitation program where seven interdisciplinary teams conducted projects aimed at improving the care of the older person in the acute sector. Aims To develop and implement a structured intervention known as the Knowledge Translation (KT) Toolkit to improve the fundamentals of care for the older person in the acute care sector. Three hypotheses were tested: (i) frontline staff can be facilitated to use existing quality improvement tools and techniques and other resources (the KT Toolkit) in order to improve care of older people in the acute hospital setting; (ii) fundamental aspects of care for older people in the acute hospital setting can be improved through the introduction and use of specific evidence-based guidelines by frontline staff; and (iii) innovations can be introduced and improvements made to care within a 12-month cycle/timeframe with appropriate facilitation. Methods Using realistic evaluation methodology the impact of a structured facilitation program (the KT Toolkit) was assessed with the aim of providing a deeper understanding of how a range of tools, techniques and strategies may be used by clinicians to improve care. The intervention comprised three elements: the facilitation team recruited for specific knowledge, skills and expertise in KT, evidence-based practice and quality and safety; the facilitation, including a structured program of education, ongoing support and communication; and finally the components of the toolkit including elements already used within the study organisation. Results Small improvements in care were shown. The results for the individual projects varied from clarifying issues of concern and planning ongoing activities, to changing existing practices, to improving actual patient outcomes such as reducing functional decline. More importantly the study described how teams of clinicians can be facilitated using a structured program to conduct practice improvement activities with sufficient flexibility to meet the individual needs of the teams. Conclusions The range of tools in the KT Toolkit were found to be helpful, but not all tools needed to be used to achieve successful results. Facilitation of the teams was a central feature of the KT Toolkit and allowed clinicians to retain control of their projects; however, finding the balance between structuring the process and enabling teams to maintain ownership and control was an ongoing challenge. Clinicians may not have the requisite skills and experience in basic standard setting, audit and evaluation and it was therefore important to address this throughout the project. In time this builds capacity throughout the organisation. Identifying evidence to support practice is a challenge to clinicians. Evidence-based guidelines often lack specificity and were found to be difficult to assimilate easily into everyday practice. Evidence to inform practice needs to be provided in a variety of forms and formats that allow clinicians to easily identify the source of the evidence and then develop local standards specific to their needs. The work that began with this project will continue - all teams felt that the work was only starting rather than concluding. This created momentum, motivation and greater ownership of improvements at local level. PMID- 21631869 TI - Organisational challenges when enabling ease of access for the elderly around a large acute hospital campus. AB - This paper describes one of several projects undertaken in a large acute care hospital under the banner of 'TOPIC7' The Older Person and Improving Care. It was conducted between January and November 2008. Although driven by the Royal Adelaide Hospital Nursing Service it was undertaken within a multidisciplinary framework. Large city-based tertiary hospitals, nearly without exception, are usually large sprawling campuses having evolved over time from buildings designed and constructed for lower populations and less traffic issues. Parking facilities may or may not be available on-site. In addition, drop-off points are located on the peripheries of the campus at often congested areas. These elements present a particular challenge to the elderly patient. Aims The purpose of this project was twofold: first, to investigate the challenges faced by older people when making their way from access points such as drop-off points and car parks through to hospital departments and second, to review potential solutions. Methods The multidisciplinary project team used a variety of knowledge translation tools and strategies to focus on areas of concern in relation to access for the elderly. They conducted a series of audits to guide practice improvement activities. This led to a reconsideration of currently planned strategies and planning of new initiatives to improve access for the elderly. Results The project was able to define clearly and prioritise the many challenges facing the older person trying to negotiate their way into hospital buildings and the barriers to them successfully reaching their destination in a safe and timely manner. At the beginning of the project it was apparent that a specific solution (an electric courtesy buggy) was in the process of being implemented. This intervention was reviewed in terms of consumer support and implementation feasibility. As a result of the comprehensive audit process, the 'courtesy buggy' was determined to not be the ideal solution and an alternative solution (wheelchair bank) is now being planned for implementation. Conclusions This project highlighted the challenges of working within a large and complex organisation with multiple key stakeholders and the need for clear pathways that clinician groups can follow to avoid duplication of effort and potential implementation of suboptimal strategies. PMID- 21631870 TI - South Africans' experiences of being old and of care and caring in a transitional period. AB - This focused ethnographic study aimed to illuminate a group of South Africans' experiences of being old and of care and caring in a transitional period. With a growing number of older people in Africa, studies on the individual experiences may help to develop care which is more sensitively based on the needs for older people in a changing Southern Africa context. Data were collected through group and individual in-depth interviews and participant observations which involved 16 individuals, aged 52-76. Data were analysed using a qualitative content analysis. The study showed two interrelated themes reflections on life and ubuntu - an orientation towards others. Findings were discussed from the viewpoint of the theory of gerotranscendence, showing similarities as well as differences, possibly due to societal and cultural differences. Shortage of formal care for older people living in poor conditions in Southern Africa, gave rise to the discussion for the need of a contextualized development of gerontological care. To enhance knowledge on the theory of gerotranscendence and develop guidelines for nursing in home-based care/community-based care in a South African context may be a first step to support older people in their process towards gerotranscendence. PMID- 21631871 TI - Food intake and nutritional status of hospitalised older people. AB - BACKGROUND AND AIMS: Disease is influenced by the nutritional status of the individual. We have assessed the relationship between nutritional status and food intake among recently hospitalised older people. METHODS: A cross-sectional study was undertaken with 240 older people in a hospital that provides care for the public and private healthcare systems. Nutritional status was classified by the MNA (Mini Nutritional Assessment) into: malnourished, risk of malnutrition and without malnutrition. Food intake was estimated by the reported food intake during a typical day. The Kruskal-Wallis test was used to compare the medians and the correlation coefficient of Spearman to verify the relationship between the consumption of energy, protein and vitamin C and MNA scores. RESULTS: 33.8% were classified as adequate regarding nutritional status; 37.1% were classified as being at risk of malnutrition and 29.1% were classified as malnourished. The malnourished individuals reported significantly less energy and nutrient intake than those at risk of malnutrition or those without malnutrition (P = 0.001). Not all nutrient intake, just some (iron, cholesterol and fibre), were lower in malnourished people. CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: Deterioration of the nutritional status of older people is accompanied by a reduction in energy and some nutrient intake. The investigation of food intake in older people could provide important information about nutritional risk. PMID- 21631872 TI - Evaluating pressure ulcer care by home healthcare nurses. AB - AIM: To field test an objective evaluation tool consisting of process-oriented quality indicators for pressure ulcer care, using nursing charts of homecare nurses. These quality indicators were developed by the authors. BACKGROUND: Most Japanese homecare nursing agencies are small and to use much of their human and economic resources to improve the care quality is not realistic. A simple and effective system for quality assurance/improvement needs to be considered. DESIGN: Descriptive study design, using the chart reviews of 34 homecare nursing clients from two homecare nursing agencies. METHODS: Nursing charts were evaluated using our quality indicators for pressure ulcer care, and whether the charts pass or not in terms of the practices described in the quality indicator was rated. The pass rates by chart and nurses' self-evaluation were compared, and pass rates by charts were examined. Results. The evaluation by chart review generally matched with self-evaluations. The pass rates by charts were higher for indicators related to wound treatment than those related to preventive care. CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: Home healthcare nurses could give more attention to pressure ulcer prevention. Regular self-checks of quality indicators may remind the nurses of the importance of prevention. PMID- 21631873 TI - Nurse-patient encounters in the hospital ward, from the perspectives of older persons: an analysis using the Authentic Consciousness Framework. AB - BACKGROUND: Despite a growing theoretical base to support the development of person-centred practice, the evidence would suggest that this way of working is somewhat elusive in the care of older people. AIMS: The study aims to explore nurse-older person encounters and relationships within the context of person centredness. DESIGN AND METHODS: This study involved secondary analysis of interview data originally collected from a commissioned study investigating whole systems approaches in services for older people. The process of analysis used the Authentic Consciousness Framework. RESULTS: Nurses are often invisible to the patient, unless they are providing care to address a physical need. A sense of rolelessness pervaded the data, when patients were deprived from actively participating in important decisions about their future care. CONCLUSIONS: Patients would like nurses to work with them in more transparent ways. Patients are very conscious that nurses are busy and attempt to share the coping with the busy workload by limiting their expectations of the nurse. RELEVANCE TO CLINICAL PRACTICE: Person-centred strategies must enhance the capacity of not only older patients and their ability to assert self, but also the capacity of their nurses. Nurses must work to actively recruit the patient in all decision making. PMID- 21631874 TI - Nursing home residents' views on dying and death: nursing home employee's perspective. AB - AIM: To reveal nursing home employees' views on dying and death among older people they cared for. BACKGROUND: Palliative care stakeholders recently included more groups in their definition of palliative care; older people constitute one such group. Consequently, palliative care systems, which will serve a large, aging cohort, will require new skills. The first stage in skills acquisition is to gather current views on dying and death. DESIGN: Qualitative descriptive study that uses focus group discussions for data collection; 20 employees in 4 Swedish nursing homes participated. METHOD: Data were analysed using qualitative content analysis. RESULTS: The following categories were conceptualised: alleviating suffering and pain; finding meaning in everyday life; revealing thoughts and attitudes about death; taking care of the dead person's body; and coping with the gap between personal ideals and reality. CONCLUSIONS: A deeper understanding of the palliative care philosophy is needed to further develop and tailor care for the dying persons in nursing homes. RELEVANCE TO CLINICAL PRACTICE: To get public support for palliative care, the silence surrounding dying and death must be broken. Employees must receive education to prepare for all aspects of their work, and management must account for employees' situation when planning the care. PMID- 21631875 TI - Online health information use by participants in selected senior centres in Korea: current status of internet access and health information use by Korean older adults. AB - AIMS AND OBJECTIVES: The purpose of this research was to describe how older adult internet users from senior centres in Korea accessed and used health information on the internet and the perceived barriers that prevented internet use. BACKGROUND: Increasingly, patients are using health-related information available on the internet. Internet based health information has influenced patient knowledge and expectations regarding healthcare. However, there are few studies about the use of internet for health information by older adults, and even less is known about older adults in Korea. DESIGN: Based on the Access to Information framework developed by McCreadie and Rice, this quantitative study with a qualitative adjunct explored the older adults' use of internet for health information. METHODS: A convenience sample of 91 community-dwelling older adults in Korea was recruited from 10 randomly selected senior centres, using the modified Kaiser Family Foundation Survey as the basis for structured interview. RESULTS: Half of older internet users (n = 13) had used online health information and found it was helpful. Barriers to internet use were: no interest (90.8%); too complicated (75%); no experience in using internet (~50%); too expensive (~50%); or disability (10%). Qualitative findings revealed that the prime reason for non use of the internet was related to literacy and level of education due to historical background. CONCLUSIONS: The results of quantitative and qualitative analysis supported that the internet users were relatively younger, more educated, male, had a higher income, received more support from others and all from large senior centres. IMPLICATIONS FOR PRACTICE: This study is important because the results can provide valuable insights into the potential for health education and health communication for older adults in Korea using the internet. It could affect health promotion, policy and education for older adults in Korea. PMID- 21631876 TI - Nutritional screening in community dwelling older adults. AB - AIMS AND OBJECTIVES: The purpose of this study was to test whether a combination of validated tools, one for each of five leading nutritional risk factors, could predict unintentional weight loss in community dwelling older adults. BACKGROUND: Non-invasive, easily administered nutritional screening tools for community dwelling older adults are few and those that are available are problematic. METHOD: Convenience samples of 115 adults >=65 were interviewed. Height, weight and measures of the five nutritional risk factors were collected at interviews 6 months apart. DESIGN: Repeated measures. RESULTS: 91 subjects completing T2 were largely white (95.6%), female (69%), well educated and in good health. Multiple regression was conducted with unintentional weight loss as the dependent variable and depression (the GDS-SF), the Lubben Social Networking Scale, food security, food intake and Independent Activities of Daily Living as the predictor variables. The regression model was statistically significant (F (5, 85) = 0.30852, P = 0.003) with an adjusted r(2) of 0.137. CONCLUSIONS: Five validated tools can be administered by nurses or non-professionals to screen for nutritional risk factors leading to unintentional weight loss. IMPLICATIONS FOR PRACTICE: Early screening has the potential to identify either nutritional risk or nutritional decline in older adults. PMID- 21631877 TI - Factors associated with nutritional risk in 75-year-old community living people. AB - AIM: To identify risk factors for being at nutritional risk, by means of a nutritional screening, in a population based sample of 75-year-old people living in three county councils in Sweden. BACKGROUND: Undernutrition in older people is known to contribute to poor health. The instrument 'Nutritional Form For the Elderly' (NUFFE) helps to identify those at nutritional risk. METHOD: The screening instrument 'Nutritional Form For the Elderly', background variables and health related questions were mail distributed. A total of 1461 persons (75 years old) were included in the study. Descriptive statistical methods were used in the analyses. RESULTS: One percent of the participants had high risk, 21.3% medium and 77.7% low risk for undernutrition. Medium or high risk was predicted by: living alone, receiving help and impaired perceived health. Low Body Mass Index was associated with low risk for undernutrition. CONCLUSION: By using a simple nutritional screening instrument, significant risk factors were highlighted. Relevance to practice. This instrument can identify older people at nutritional risk and is easy to use. Older people living alone have an increased risk of undernutrition. Body Mass Index (BMI) should be used with caution as one and only indicator of nutritional risk in older people. PMID- 21631878 TI - Older orthopaedic patients' perceptions of individualised care: a comparative survey. AB - AIM AND OBJECTIVES: To describe and compare the individualised care perceptions of older orthopaedic patients' and patients of working age. BACKGROUND: Age has been found to influence perceptions of care and although individualised care is highlighted in the literature, it is seldom studied from an older person's perspective. DESIGN: Descriptive and comparative. METHODS: Data were collected using the Individualised Care Scale from orthopaedic patients (n = 420, response rate 84%). The participants were divided into two groups: those 65 and over (n = 149) and those under 65 and working (n = 271). Data analysis used descriptive and inferential statistics. RESULTS: Patients expressed a desire for individualised care, and gave relatively good evaluations about the perceived support for their individuality and the realisation of individualised care. Differences in the perceptions of individualised care were found between, but not within, the two groups. The older patients were more positive in their evaluations. CONCLUSIONS: There is a need for programmes of individualised care that are age-adjusted. RELEVANCE TO CLINICAL PRACTICE: As the older population rises worldwide individualised care becomes more important in the care of older people. These findings provide baseline data for the development of individualised nursing care from the patients' perspective. PMID- 21631879 TI - Restraints or alternatives: safety work in care of older persons. AB - AIM: To examine the factors and complexity that account for physical restraints use and their alternatives as nurses' protect the safety of older adults. BACKGROUND: Research on physical restraints has repeatedly shown the human, financial and work costs associated with their use, but their use continues to be a major quality issue across hospitals and across countries for older persons. METHOD: Interviews with medical surgical nurses in a Midwestern hospital in the United States were recorded and interpreted using grounded theory. RESULTS: Using data that described the complexity of initiating, maintaining, or discontinuing restraints, we captured the density of these processes and showed how nurses made decisions about restraint use. The tasks carried out to protect the older person's safety often involved using restraints or their alternatives and these tasks then set in motion a cascade of other types of tasks or work, e.g., diagnostic, comfort, articulation. CONCLUSIONS: The study extended the work on chronic illness trajectories and formulated a framework for understanding the complexity of the use of restraints and their alternatives. Using a tracing approach we were able to predict the types of situations that would most likely result in the initiation, maintenance and discontinuation of restraints or the use of alternative procedures. RELEVANCE TO PRACTICE: The study has identified a process of 'safety work' that identified the risk associated with the problem, the work to cope with the problem and risk, the conditions that influenced the process and the consequences or outcomes of the work strategies to maintain the older person's safety. PMID- 21631880 TI - Association between self-reported care needs and the allocation of care in Norwegian home nursing care recipients. AB - OBJECTIVES: This study investigated the relationship between patients' self reported illness, daily afflictions, and the frequency of home nursing care, and whether patients' coping resources influenced the allocation of care. DESIGN, SAMPLE AND MEASUREMENTS: A cross-sectional survey was adopted. Two hundred and forty-two people aged 75 years and above receiving home nursing care participated in the study. Binary logistic regression model was used to test the effects of the independent variables on home nursing care. RESULTS: Poor capacity to perform activities of daily living and high level of education were directly associated with a high frequency of home nursing care. Lack of perceived social support affected the amount of home nursing care allocated only when feelings of loneliness were connected with poor activities of daily living functioning. Interaction effects revealed that perceived social support influenced the amount of home nursing care in persons with higher education, in persons with low education, no such association were found. No associations were found between coping resources and home nursing care. CONCLUSIONS: Impaired capacity to perform activities of daily living was the main reason for care allocation. Education was associated with more formal care. Patients with low perceived social support combined with a low education level was a particularly vulnerable group. PMID- 21631881 TI - The importance of 'approaching' older people: a grounded theory. AB - AIMS AND OBJECTIVES: The aim of this study was to identify and describe the ethical values in caring encounters as experienced by older patients in their daily interaction with nurses in wards for older people. BACKGROUND: Ethical values and morals are important aspects that influence the quality of care. METHODS: Empirical observational study including follow-up interviews. Twenty-two older patients participated voluntarily in this study. Constant comparative analysis, the core foundation of grounded theory was used. RESULTS: Five categories: being addressed, receiving respect, desiring to participate, increasing self-determination and gaining self-confidence formed the bases for the core category. APPROACHING: Approaching concerns how people become closer to each other in a physical space. It also includes how people become closer to each other in a dialogue, involving verbal or bodily communication. CONCLUSIONS AND RELEVANCE TO CLINICAL PRACTICE: Approaching indicates the ethical values that guide nurses in their caring encounters with older patients. These values are noted by the patient and have an individual value as well as leading to improved quality of their care. The older patient will be confident and satisfied with the caring encounter if the desired components in the nurse's approaching are exhibited. PMID- 21631882 TI - Upstream thinking and health promotion planning for older adults at risk of social isolation. AB - AIMS AND OBJECTIVES: To raise awareness of social isolation, and provide an approach to first conceptualise and then prevent social isolation among older community-dwelling persons. BACKGROUND: Older adults comprise a vulnerable population for social isolation and its associated health risks. DESIGN: Literature review. METHODS: Canada's Population Health Promotion Model was chosen as a comprehensive tool to understand and prevent social isolation. Research studies were sought to identify key health determinants and evidence-based options for preventing social isolation. RESULTS: Around 1 out of 6 older persons are socially isolated and three health determinants are of prime importance: (i) income and social status; (ii) personal health practices and coping skills and (iii) social support networks. Evidence-based interventions targeted to these health determinants are suggested. CONCLUSION: Nurses are a key group to advocate for actions needed to prevent social isolation. IMPLICATIONS FOR PRACTICE: Nurses can play a vital role in minimising social isolation through a variety of educational, prevention and political lobbying activities. PMID- 21631884 TI - Evaluating the effectiveness of the abilities-focused approach to morning care of people with dementia. AB - BACKGROUND AND PURPOSE: The abilities-focused approach demonstrated efficacy in promoting engagement of residents with dementia in care. The extent to which these resident outcomes can be replicated when the abilities-focused approach is implemented by nursing staff under the conditions of day-to-day practice was investigated in this study. The aim was to examine changes in resident outcomes before and after nursing staff' implementation of the abilities-focused approach and the contribution of this approach to resident outcomes. METHODS: A one-group pretest-post-test design was used. Observational data were obtained from 65 residents with dementia. The data pertained to the implementation of abilities focused strategies during episodes of morning care and residents' level of agitation, participation in morning care and physical and psychosocial functioning. RESULTS: The number of abilities-focused strategies used during morning care increased at post-test. No clinically important changes in resident outcomes were observed over time. However, the implementation of some strategies was associated with residents' participation in care and functioning at post test. The results provided preliminary evidence of the effectiveness of the abilities-focused approach in maintaining functioning in people with dementia. Additional research is needed to understand the mechanism underlying the effects of this person-centred approach to care on resident outcomes. IMPLICATIONS FOR PRACTICE: Implementation of the abilities-focused approach assists people with dementia to participate in morning care. PMID- 21631883 TI - Efficacy of the use of evidence-based algorithmic guidelines in the acute care setting for pain assessment and management in older people: a critical review of the literature. AB - AIM: The aim of the literature review was to identify all examples of primary research using an algorithmic approach for the implementation of a clinical practice guideline relating to pain assessment and/or management within acute care, with a specific focus on older people. DESIGN: Critical literature review. DATA SOURCES: Inclusion criteria were; English language publications within the past 13 years; peer reviewed; research conducted within a hospital; about adult inpatients. Exclusion criteria; research located outside of a hospital context; quality improvement studies; rehabilitation studies and literature reviews. REVIEW METHODS: Critical appraisal of the literature by using a qualitative interpretation of a translational approach. The literature was thematically mapped according to the criteria of credibility, transferability, plausibility and applicability. RESULTS: No clinical practice guideline was found that directly related to both assessment and management of pain using an algorithm in acute care for older people. Five studies were found to have relevance and were critically evaluated. CONCLUSION: A critique of the literature shows that an algorithmic approach is feasible for translation into a clinical practice guideline for assessment and management of pain in older people within the acute care setting. IMPLICATIONS FOR PRACTICE: Implementation of any algorithmic approach requires consideration of the environment, culture and availability of resources. PMID- 21631885 TI - Older adults' knowledge of pressure ulcer prevention: a prospective quasi experimental study. AB - AIM: To test an evidence base patient education leaflet to evaluate older adults' knowledge of pressure ulcers and prevention strategies. BACKGROUND: The increasing population of older adults living in the community managing chronic health conditions are at risk of pressure ulcer development. Education leaflets are a useful adjunct to reinforce healthcare professional's verbal information to promote healthy lifestyles choices. However, little is known of the effectives of pressure ulcer prevention educational leaflets for older adults. METHODS: A quasi experimental uncontrolled pre-test, post-test study of participants' knowledge of pressure ulcer and preventative strategies was conducted. Community dwelling older adults (n = 75) were recruited to this study. Older adult's knowledge was measured pre- and postdistribution of an education intervention. A risk assessment scale was recorded to identify whether this cohort of older adults were actually at risk of developing pressure ulcers. CONCLUSION: The results indicate that an education leaflet enhanced patients' knowledge relating to pressure ulceration. Printed education materials increase knowledge and understanding which may lend to older adults adopting healthy behaviours. IMPLICATIONS FOR PRACTICE: An education leaflet can help older adults and their carers to be more empowered as active participants in reducing the incidence of pressure ulceration. PMID- 21631886 TI - Use of the resiliency model of family stress, adjustment and adaptation in the analysis of family caregiver reaction among families of older people with congestive heart failure. AB - BACKGROUND: Little attention has been given to the resiliency process of family caregivers for older people hospitalized with congestive heart failure. AIMS: The purpose of this study was to examine the influences of older peoples' activities of daily living dependency, family caregivers' spiritual well-being, quality of relationship, family support, coping and care continuity on the burden of family caregivers of hospitalized older people with congestive heart failure using the Resiliency Model of Family Stress, Adjustment, and Adaptation. DESIGN AND SAMPLE. A descriptive, correlational research design was used. There were 50 family caregivers and 50 older people diagnosed with congestive heart failure recruited from medical and surgical units of two Midwest medical centres. METHODS: Data on activities of daily living dependence of older people, family caregiver burden, spiritual well-being, coping strategies, quality of older people-caregiver relationship and care continuity were collected using structured questionnaires. RESULTS: The findings indicated that there were significant positive associations between patients' activities of daily living dependence and family caregiver burden and between lack of family support and family caregiver burden. There were significant negative associations between quality of relationship and family caregiver burden, between care continuity and family caregiver burden, between coping and family caregiver burden and between spiritual well-being and burden. In hierarchical multiple regression, the model variables accounted for 66% of the variance in family caregiver burden. Patients' activities of daily living dependence, quality of relationship and lack of family support are significant predictors of family caregiver burden. CONCLUSIONS: The best predictors of family caregiver burden, using the resiliency model, were the older persons' activities of daily living dependence, family support, quality of relationship, care continuity, coping strategies and spiritual well-being. Implications for practice. It is vital for nurses to assess family caregivers' needs and resources and the quality of the older person-family caregiver relationship in developing a plan of care that reduces family caregiver burden. PMID- 21631887 TI - Nurse champions of older people: a national audit and critical review. AB - AIMS: This paper aims to equip nurses working with older people with an understanding of the roles and activities of Champions for Older People. BACKGROUND: The position of Older People's Champion was established in England to help implement the National Service Framework for Older People at local level. METHODS: This paper reports on the findings of a national survey of older people's Champions. It sets this in the context of developments around health polices for older people. FINDINGS: Forty-three of the 209 Champions who participated in this study were nurses. Nurses were the most commonly appointed clinical Champions in National Health Service Trusts responding to the survey; they used this position to promote changes within their organisation and locality. While focusing on the implementation of National Service Framework for Older People targets and requirements, some used their position to work with organisations of local older people and advocated for older patients' interests. CONCLUSION: Nurses conceived their position as clinical Champions broadly and reported that they used this role to promote service improvements for older people. Those who established networks with other Champions and older people's groups found this useful. Their role as Champions appeared to encourage a proactive and outward-looking perspective and provides evidence of the workings of one model of Championship. IMPLICATIONS FOR PRACTICE: Embedding championing roles within organisations may be one way of bringing about change in the way that services to older people are developed. PMID- 21631888 TI - Gerontological rehabilitation nursing of older patients in acute health centre hospitals: nursing views. AB - AIM: The aim was to describe gerontological rehabilitation nursing in an acute hospital setting from nursing staff's points of view. BACKGROUND: In the model of gerontological rehabilitation nursing, older people are active operators in their own rehabilitation process. It is the task of nursing staff, together with the patient, their relatives and a multiprofessional team, to support the patient's commitment to the rehabilitation goal. METHODS: In this study, a cross-sectional design was used. Registered nurses (RNs) and practical nurses (PNs) (n = 367) from four different hospitals responded to the questionnaire. Data analysis was carried out with spss using descriptive statistics. RESULTS: The majority of nursing staff (66%) considered that older rehabilitation patients thought that rehabilitation was activity undertaken by physiotherapists aimed at improving patients' functional abilities. The wards in question have well-functioning rehabilitation teams. The rehabilitation of older patients is a goal-oriented activity. Despite this, not everyone was aware of the goals. Essential patient care and promotion of rehabilitation were well attended to by nurses. Emotional support was emphasised in maintaining the motivation of older people. CONCLUSIONS: Because RNs and PNs, unlike other professionals, see older rehabilitation patients 24 hours a day and are able to assess their possibilities of coping independently with essential tasks, nursing staff could have an active role in the rehabilitation team. The education of nursing staff must focus on boosting self-esteem and teaching independent decision-making in promoting the rehabilitation of older persons and the assessment of their progress. IMPLICATIONS FOR PRACTICE: Nurse managers must arrange further education for nurses in promoting the rehabilitation of older persons. Nurse managers must also arrange time for the multiprofessional team to discuss and agree joint rehabilitation goals. PMID- 21631889 TI - 'Finding home': a grounded theory on how older people 'find home' in long-term care settings. AB - BACKGROUND: A link between residents 'feeling at home' in long-term care facilities and 'quality of life' is emerging in the literature. Few studies, however, have focused on what helps residents to find a home in long-term care settings. This study aimed to fill this gap. AIM: This study aimed to understand older peoples' perceptions of 'being at home' in long-term care settings and the factors that influence these perceptions. DESIGN: Grounded theory guided the study design. Residents (n = 61) living in public or private long-term care settings were interviewed using unstructured interviews. FINDINGS: Four categories were identified as critical to finding a home in long-term care settings: 'continuity', 'preserving personal identity', 'belonging' and 'being active and working'. 'Finding Home' was conceptualised as the core category. The potential to 'find home' was influenced by mediating and facilitating/constraining factors. CONCLUSIONS: The Theory of Finding Home was generated from the data. This theory describes the factors critical to 'finding home' in long-term care settings. IMPLICATIONS FOR PRACTICE: The Theory of Finding Home gives insight into what matters to older people living in long-term care settings. Strategies to help generate a feeling of home in long-term care settings are shared. PMID- 21631890 TI - Identifying and handling abused older clients in community care: the perspectives of nurse managers. AB - BACKGROUND: Abuse of older people can lead to an increased need for health care services, and early identification of and intervention with abused, older clients should be a priority. OBJECTIVES: To gain knowledge about how nurse managers and their staff identify and handle abused older clients. DESIGN: Qualitative study. METHODS: Fifty-two participants were interviewed, the text transcribed and analysed using manifest qualitative content analysis. FINDINGS AND DISCUSSION: The identification and handling of older victims depended on individual nurses and the involvement of the participants and are based on clinical experience and less on knowledge through professional training and education. The participants emphasised the need for an individualised approach to older victims suspected of abuse. The important role of the nurse managers in the handling of abuse is discussed as well as the need for support of the nurse managers and the staff in community care by increased interdisciplinary collaboration to handle complex cases. CONCLUSION: The identification and handling of abused older clients are a challenging issue for community care, and more knowledge and support are needed. IMPLICATIONS FOR PRACTICE: The concept of elder abuse must be clarified, and strategies for its identification and intervention must be developed. PMID- 21631891 TI - Evaluation of the nucleated red blood cell count in neonates using the Beckman Coulter UniCel DxH 800 analyzer. AB - INTRODUCTION: Nucleated red blood cell (NRBC) count is closely associated with the prognosis of neonates. The analysis of NRBC has traditionally been measured manually. Recently, a newly developed automated hematology analyzer, the UniCel DxH 800 (DxH 800), was released. The goal of our study was to evaluate the performance of the DxH 800 NRBC method in neonates with the reference manual method and against previous generation hematology analyzers. METHODS: NRBCs were counted in 162 neonatal blood samples using the DxH 800, LH 750, and ADVIA 2120 vs. the reference manual technique. The concordance rate, sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were obtained. RESULTS: The DxH 800 showed an R(2) value of 0.945 and the concordance rate of 93.8%. Further assessment revealed 85.3% sensitivity, 96.1% specificity, 85.3% PPV, and 96.1% NPV, resulting in the highest area under the curve (0.961). The LH 750 and ADIVA 2120 demonstrated R(2) values of 0.851 and 0.529, respectively. CONCLUSION: The results obtained indicate the UniCel DxH 800 to be an excellent test on neonatal blood and superior to the other analyzers. Therefore, the DxH 800 is an effective and highly sensitive system for the analysis of NRBCs on newborns. PMID- 21631892 TI - Precise quantification of haemoglobin in erythroid precursors and plasma. AB - INTRODUCTION: Haemoglobin (Hb) quantification in whole blood is possible by various spectrophotometric methods. However, determination of low-level Hb in erythroid precursors or haemolytic plasma is inaccurate because of contribution from light scatter and/or nonhaemoglobin components with overlapping absorbance. Therefore, this study developed a sole method allowing accurate spectrophotometric quantification of Hb at a low concentration range. METHODS: Advantage was taken of the unique absorption spectra of carbon monoxide-Hb complex (COHb) as compared to oxyHb. The visible absorption spectra of samples were recorded prior and following carbon monoxide exposure. A difference extinction coefficient at the maximal difference absorption was used to calculate Hb concentrations. RESULTS: Known amounts of Hb were added to mouse erythroleukaemia (MEL) cells lysate or plasma to yield known 'theoretical' concentrations. The concentrations were measured by the current and known methods. The current method was found much more accurate compared with previous methods specifically at low concentrations. CONCLUSION: The method is valid for accurate quantification of Hb at a wide concentration range (>0.1 MUm/L) in erythroid precursors or plasma and is optional for other biological fluids. PMID- 21631893 TI - Metformin as an antitumor agent in cancer prevention and treatment. AB - Recent epidemiological investigations conducted in diabetic cohorts and cancer patients have found that metformin users have lower risks for cancer than those using insulin or insulin secretagogues. Studies conducted in various animal tumor models and cancer cell lines have demonstrated that metformin prevents tumor development or inhibits cell proliferation. In addition, a recent clinical trial has shown that short-term use of metformin reduces aberrant crypt foci (ACF) formation in non-diabetic patients with ACF. The antitumor activity of metformin may be mediated through its regulatory effect on hormonal, metabolic, and immune functions. Metformin achieves glycemic control by reducing hepatic glucose production and increasing the muscle intake of glucose, thus lowering levels of circulating glucose and, consequently, insulin. The major molecular targets of metformin are the liver kinase B1 (LKB1)-AMP-activated protein kinase (AMPK) signaling and mammalian target of rapamycin (mTOR) pathways, which are central in the regulation of cellular energy homeostasis and play a crucial role in the control of cell division and cell proliferation. Metformin has been shown to improve endothelial function, decrease inflammatory activity, and regulate immune function. Increasing experimental evidence provides a strong biological rationale for metformin as an antitumor and chemopreventive agent. Metformin is being tested as an adjuvant cancer therapy in clinical settings, and metformin is recommended for all cases of Type 2 diabetes without contraindications. As described in this review, the chemopreventive value of metformin is not restricted to diabetic or obese individuals. PMID- 21631894 TI - Strategies to detect abnormal glucose metabolism in people at high risk of cardiovascular disease from the ORIGIN (Outcome Reduction with Initial Glargine Intervention) trial population. AB - BACKGROUND: To investigate whether the combination of HbA1c and fasting plasma glucose (FPG) can be used for the diagnosis of diabetes and impaired glucose tolerance (IGT) in people at high risk of cardiovascular disease (CVD). METHODS: A cross-sectional study was performed on 2907 people at high risk of cardiovascular events but without a previous diagnosis of diabetes. Optimal cut off points and the diagnostic potential of FPG, HbA1c, and their combination were determined. RESULTS: The sensitivity of the usually applied FPG cut-off point of 7.0 mmol/L to diagnose diabetes mellitus was low (59.0%). Receiver operating characteristic (ROC) curve analysis indicated that the optimal cut-off points for the diagnosis of diabetes using FPG and HbA1c were 6.4 mmol/L (sensitivity 75.7%; specificity 77.5%; likelihood ratio 3.37) and 5.9% (41 mmol/mol; sensitivity 68.7%; specificity 67.1%; likelihood ratio 2.09), respectively. To diagnose IGT, the optimal cut-off points for FPG and HbA1c were 6.1 mmol/L (sensitivity 57.1%; specificity 57.9%) and 5.7% (39 mmol/mol; sensitivity 63.8%; specificity 60.3%), respectively. For diabetes, combining cut-off points for FPG and HbA1c identified four categories with likelihood ratios ranging from 5.59 to 0.21, and post-test probabilities between 69.3% and 7.8%. For IGT, likelihood ratios varied between 2.05 and 0.56, whereas post-test probabilities ranged from 84.0% to 58.8%. CONCLUSIONS: Using FPG alone results in the underdiagnosis of glucometabolic abnormalities in people at high risk of CVD. Using an algorithm with both HbA1c and FPG improves the detection of diabetes, but not IGT, and could be easily implemented in patient care. PMID- 21631895 TI - Autologous islet cell transplantation to prevent surgical diabetes. AB - Autologous islet transplantation (AIT) is performed to prevent surgical diabetes after total or semi-total pancreatectomy for the treatment of chronic pancreatitis with severe abdominal pain. In addition, AIT is used in cases of benign pancreatic tumors and pancreatic trauma. It has been shown that AIT results in better outcomes in terms of glycemic control compared with allogeneic islet transplantation. The reasons for the favorable outcomes of AIT are thought to be: (i) patients have no autoimmune diseases; (ii) the transplanted islets do not suffer allogeneic rejection; (iii) diabetogenic antirejection drugs are not required; (iv) pancreata do not undergo a cytokine storm as a result of periods of brain death; (v) the period of cold preservation of retrieved pancreata is short; (vi) the isolated islets are immediately transplanted without culture; and (vii) pancreata with pancreatitis may contain more progenitor cells. Further research into AIT would help improve the results of allogeneic islet transplantation. Conversely, the technical difficulties associated with islet isolation appear to be the largest hurdle for AIT; therefore, remote center islet isolation may prove to be key in the promotion of this treatment. PMID- 21631896 TI - Review of the effects of the traditional Chinese medicine Rehmannia Six Formula on diabetes mellitus and its complications. AB - Rehmannia Six Formula (RF) is a formula that is commonly used in Traditional Chinese Medicine to treat patients with diabetes. A literature search was performed in PubMed for the years 2000-2009 using the key words RF, Rehmannia glutinosa, Fructus Corni, Dioscorea sp. (D. alata, D. opposita, D. batatas), Poria cocos, Alisma sp. (A. orientalis, A. plantago aquatica), and Paeonia suffruticosa/Cortex Moutan. On the basis of the publications found, RF appears to have beneficial effects on blood glucose, neuropathy, and nephropathy. There is also evidence of anti-inflammatory and antioxidant effects. Although there are many studies on compounds extracted from individual herbs, there are not many studies on RF as a whole. Because there is preliminary evidence that RF may be a useful supplement for the prevention of diabetic complications, clinical studies are warranted. For future clinical studies, it is recommended that details are provided regarding the preparation of RF and that the ratio of the individual components in RF is standardized so that results across studies can be compared. PMID- 21631897 TI - Replication and further characterization of a Type 1 diabetes-associated locus at the telomeric end of the major histocompatibility complex. AB - BACKGROUND: We recently reported an association between Type 1 diabetes and the telomeric major histocompatibility complex (MHC) single nucleotide polymorphism (SNP) rs1233478. As further families have been analyzed in the Type 1 Diabetes Genetics Consortium (T1DGC), we tested replication of the association and, with more data, analyzed haplotypic associations. METHODS: An additional 2717 case and 1315 control chromosomes have been analyzed from the T1DGC, with human leukocyte antigen (HLA) typing and data for 2837 SNPs across the MHC region. RESULTS: We confirmed the association of rs1233478 (new data only: P=2.2E-5, OR=1.4). We also found two additional SNPs nearby that were significantly associated with Type 1 diabetes (new data only rs3131020: P=8.3E-9, OR=0.65; rs1592410: P=2.2E-8, OR=1.5). For studies of Type 1 diabetes in the MHC region, it is critical to account for linkage disequilibrium with the HLA genes. Logistic regression analysis of these new data indicated that the effects of rs3131020 and rs1592410 on Type 1 diabetes risk are independent of HLA alleles (rs3131020: P=2.3E-3, OR=0.73; rs1592410: P=2.1E-3, OR=1.4). Haplotypes of 12 SNPs (including the three highly significant SNPs) stratify diabetes risk (high risk, protective, and neutral), with high-risk haplotypes limited to approximately 20,000 bp in length. The 20,000-bp region is telomeric of the UBD gene and contains LOC729653, a hypothetical gene. CONCLUSIONS: We believe that polymorphisms of the telomeric MHC locus LOC729653 may confer risk for Type 1 diabetes. PMID- 21631899 TI - Stratifying and tailoring HbA1c control targets for adults with Type 2 diabetes: interpretation of the consensus proposed by the Chinese Society of Endocrinology. AB - Glycemic control is an important goal of treatment to delay the progression of and complications associated with diabetes, but controversies exist regarding individual HbA1c control targets for different patients. With the aim of optimizing outcomes and minimizing adverse events, a preliminary consensus on HbA1c control targets for adults with Type 2 diabetes has been proposed by the Chinese Society of Endocrinology (CSE). Instead of recommending a general standard value for all patients, the CSE suggests that a relatively reasonable stratified and tailored target for individual patients should take into consideration both clinical status and social factors. Principles governing the establishment of a glycemic control target include safety, feasibility, scientific evidence, and customized care, of which the most important factor is safety. In addition to controlling plasma glucose, equal consideration should be given to other vascular disease risk factors. PMID- 21631900 TI - Liver enzymes, metabolic syndrome, and insulin resistance. PMID- 21631898 TI - Apelin and insulin resistance: another arrow for the quiver? AB - Apelin is a newly discovered peptide hormone that has recently been linked to insulin resistance and obesity. Data collected from both the clinical and basic research settings show that apelin: (i) is correlated with the states of insulin resistance and obesity; (ii) stimulates glucose utilization; (iii) decreases insulin secretion; and (iv) negatively regulates catecholamine-mediated lipolysis. These and other lines of evidence demonstrate that apelin may be a potentially viable candidate in the search for treatments for Type 2 diabetes and the insulin resistance (metabolic syndrome). The present review summarizes the literature on the regulation by apelin of glucose and lipid metabolism and the signaling pathways involved. PMID- 21631901 TI - Assessing the risk of lower extremity amputations using an administrative data based foot risk index in elderly patients with diabetes. AB - BACKGROUND: The aims of the present study were to derive a foot risk index using administrative data similar to the International Working Group on the Diabetic Foot clinical risk classification scheme and to evaluate its association with the risk of initial lower extremity (through foot or above) amputation (ILEA). METHODS: Merged Veterans Health Administration administrative and Medicare inpatient and outpatient claims data for a historical cohort of Veterans Health Administration users with diabetes were analyzed. Individuals with diabetes, aged >=67 years in 1999, were classified into seven foot risk categories (where 0=no foot risk conditions; and 6=severe foot conditions). The outcome variable was the incidence of an ILEA over the period 2000-2004. Foot risk was derived from 1998 to 1999, and other independent variables were derived from 1999. The associations between foot risk category and risk of ILEA were evaluated using Cox proportional hazards regression models. RESULTS: Of 255,534 individuals with diabetes, 54.8% had a documented foot condition in 1999 and 6869 (26.9 per 1000) had an ILEA over the period 2000-2004. We found increased rates of ILEAs as foot risk increased. Based on a Cox regression model containing all independent variables, the hazard ratios for risk of amputation increased progressively from 1.13 (95% CI 1.00 1.28) for the lowest foot risk category to 6.75 (95% CI 6.31-7.23) for the highest foot risk category, compared with individuals without defined foot conditions. CONCLUSIONS: An administrative data-derived foot risk category was associated with risk of ILEA in a risk level-dependent pattern. PMID- 21631902 TI - Five Ms of adherence. PMID- 21631903 TI - First insulinization with basal insulin in patients with Type 2 diabetes in a real-world setting in Asia. AB - BACKGROUND: The First Basal Insulin Evaluation (FINE) Asia study is a multinational, prospective, observational study of insulin-naive Type 2 diabetes mellitus (T2DM) patients in Asia, uncontrolled (A1c >= 8%) on oral hypoglycemic agents, designed to evaluate the impact of basal insulin initiation. METHODS: Basal insulin was initiated with or without concomitant oral therapy and doses were adjusted individually. All treatment choices, including the decision to initiate insulin, were at the physician's discretion to reflect real-life practice. RESULTS: Patients (n=2679) from 11 Asian countries were enrolled (mean [+/-SD] duration of diabetes 9.3+/-6.5 years; weight 68.1+/-12.7 kg; A1c 9.8 +/- 1.6%). After 6 months of basal insulin (NPH insulin, insulin glargine, or insulin detemir), A1c decreased to 7.7+/-1.4%; 33.7% patients reached A1c <7%. Fasting blood glucose (FBG) decreased from 11.7+/-3.6 to 7.2+/-2.5 mmol/L and 36.8% of patients reached FBG <6.1 mmol/L. The mean daily insulin dose prescribed increased marginally from 0.18 to 0.23 U/kg per day at baseline to 0.22-0.24 U/kg per day at Month 6. Mean changes in body weight and reported rates of hypoglycemia were low over the duration of the study. CONCLUSIONS: Initiation of insulin therapy is still being delayed by approximately 9 years, resulting in many Asian patients developing severe hyperglycemia. Initiating insulin treatment with basal insulin was effective and safe in Asian T2DM patients in a real-world setting, but insulin needs may differ from those in Western countries. PMID- 21631904 TI - Low serum total bilirubin concentrations are associated with increased prevalence of metabolic syndrome in Chinese. AB - BACKGROUND: To investigate the association between serum concentrations of total bilirubin (TBil) in the physiological range and metabolic syndrome (MS) in middle aged and elderly Chinese, as well as any associations between serum TBil concentrations and insulin resistance, hyperinsulinemia, and systemic inflammation. METHODS: A cross-sectional study was conducted on 1423 individuals recruited from an urban community of Shanghai (average age 62.3 years) to investigate the relationship between bilirubin and cardiovascular diseases. Metabolic syndrome was defined according to the National Cholesterol Education Program Adult Treatment Panel III criteria for Asian Americans. RESULTS: Total bilirubin concentrations were significantly lower in individuals with MS compared with those without (0.65 +/- 0.21 vs 0.69 +/- 0.22 mg/dL, respectively; P=0.002). The adjusted mean concentration of TBil decreased gradually with an increase in the number of components of MS (P(trend) < 0.0001). After adjustment for a range of potential confounders (e.g. age, sex, body mass index, smoking, alcohol intake, homeostasis model assessment of insulin resistance etc.), each 1 SD increase in TBil was found to be associated with a 17% reduction in the risk of MS (odds ratio 0.83; 95% confidence interval 0.73-0.95; P=0.006). Furthermore, after adjustment for all covariables, each 1 SD increase in TBil was found to be associated with lower odds of central obesity, hypertriglyceridemia, low high density lipoprotein-cholesterol, and hyperglycemia. Serum TBil concentrations were inversely associated with hyperinsulinemia, insulin resistance, and systemic inflammation. CONCLUSIONS: Serum TBil concentrations within the physiological range were inversely associated with MS and insulin resistance, hyperinsulinemia, and systemic inflammation in middle-aged and elderly Chinese. PMID- 21631905 TI - Outer retinal cysts in age-related macular degeneration. AB - PURPOSE: To describe novel cystic structures ('outer retinal cysts' or ORC) found in the outer retina in age-related macular degeneration (AMD). METHODS: One hundred and seventy-three consecutive eyes of 88 AMD patients were prospectively examined with spectral domain optical coherence tomography (SD-OCT). The prevalence of ORCs was searched, and their sizes and shapes were determined. RESULTS: SD-OCT revealed round or ovoid, intraretinal, hyporeflective cystic structures with a hyperreflective border in 60 eyes (56%) with neovascular AMD and in six eyes (21%) with atrophic AMD. These cystic structures were of different sizes and shapes. They remained stable in all the patients after a follow-up period of 6 months. CONCLUSIONS: Outer retinal cyst is a new type of cystic structure recently identified in AMD patients. ORCs should not be confused with intraretinal exudates or cystoid cavities and therefore do not require any treatment. The histopathological nature of ORC remains to be determined. Further studies are necessary to determine their true origin. PMID- 21631906 TI - Future medical applications of single-cell sequencing in cancer. AB - Advances in whole genome amplification and next-generation sequencing methods have enabled genomic analyses of single cells, and these techniques are now beginning to be used to detect genomic lesions in individual cancer cells. Previous approaches have been unable to resolve genomic differences in complex mixtures of cells, such as heterogeneous tumors, despite the importance of characterizing such tumors for cancer treatment. Sequencing of single cells is likely to improve several aspects of medicine, including the early detection of rare tumor cells, monitoring of circulating tumor cells (CTCs), measuring intratumor heterogeneity, and guiding chemotherapy. In this review we discuss the challenges and technical aspects of single-cell sequencing, with a strong focus on genomic copy number, and discuss how this information can be used to diagnose and treat cancer patients. PMID- 21631907 TI - Should persons with autosomal dominant AD be included in clinical trials? Authors' response. PMID- 21631908 TI - Intravenous magnesium in subarachnoid hemorrhage. PMID- 21631909 TI - Care bundles: implementing evidence or common sense? AB - Care bundles aim to improve standard of care and patient outcome by promoting the consistent implementation of a group of effective interventions. However, a variety of barriers prevent their full application in clinical practice. Here, we discuss some of the benefits and limitations of care bundles in the delivery of safer and more effective and consistent health care. PMID- 21631910 TI - Concurrent analysis of choice and control in childbirth. AB - BACKGROUND: This paper reports original research on choice and control in childbirth. Eight women were interviewed as part of a wider investigation into locus of control in women with pre-labour rupture of membranes at term (PROM). METHODS: The following study uses concurrent analysis to sample and analyse narrative aspects of relevant literature along with these interviews in order to synthesise a generalisable analysis of the pertinent issues. The original PROM study had found that women experienced a higher degree of control in hospital, a finding that appeared at odds with contemporary notions of choice. However, this paper contextualises this finding by presenting narratives that lucidly subscribe to the dominant discourse of hospital as the safest place to give birth, under the premise of assuring a live healthy baby irrespective of their management type. RESULTS: This complex narrative is composed of the following themes: 'perceiving risk', 'being prepared', 'reflecting on experience', maintaining control' and relinquishing control'. These themes are constructed within and around the medical, foetocentric, risk averse cultural context. Primary data are presented throughout to show the origins and interconnected nature of these themes. CONCLUSIONS: Within this context it is clear that there is a highly valued role for competent health professionals that respect, understand and are capable of facilitating genuine choice for women. PMID- 21631912 TI - A dual role for microglia in promoting tissue inhibitor of metalloproteinase (TIMP) expression in glial cells in response to neuroinflammatory stimuli. AB - BACKGROUND: By neutralizing the effect of the matrix metalloproteinases (MMPs), the tissue inhibitors of matrix metalloproteinases (TIMPs) play a critical role in maintaining tissue proteolysis in balance. As the major reactive glial cell types in the central nervous system (CNS), microglia and astrocytes play fundamental roles in mediating tissue breakdown and repair. As such, it is important to define the TIMP expression profile in these cells, as well as the mechanisms of regulation by neuroinflammatory stimuli. METHODS: Primary mixed glial cultures (MGC), pure microglia, and pure astrocytes were used in this study. To study astrocytes, we employed a recently described pure astrocyte culture system, which has the major advantage of totally lacking microglia. The three different types of culture were treated with lipopolysaccharide (LPS) or individual cytokines, and cell culture supernatants assayed for TIMP-1 or TIMP-2 protein expression by western blot. RESULTS: LPS induced TIMP-1 expression in MGC, but not in pure astrocyte or microglial cultures. When pure astrocytes were treated with the cytokines IL-1beta, IFN-gamma, TNF or TGF-beta1, only IL-1beta induced TIMP-1 expression. Significantly, astrocyte TIMP-1 expression was restored in LPS-treated astrocyte cultures after the addition of microglia, or conditioned medium taken from LPS-activated microglia (MG-CM). Furthermore, this effect was lost after depletion of IL-1beta from MG-CM. By contrast, TIMP-2 was constitutively expressed by astrocytes, whereas microglia expressed TIMP-2 only after exposure to serum. CONCLUSIONS: Taken together, these results demonstrate an important concept in glial interactions, by showing that microglia play a central role in regulating glial cell expression of TIMPs, and identify microglial IL-1beta as playing a key role in mediating microglial-astrocyte communication. PMID- 21631913 TI - Contraception in HIV-positive female adolescents. AB - Sexual behavior of HIV-positive youths, whether infected perinatally, through risky behavior or other ways, is not substantially different from that of HIV uninfected peers. Because of highly active antiretroviral therapy, increasing number of children, infected perinatally, are surviving into adolescence and are becoming sexually active and need reproductive health services. The objective of this article is to review the methods of contraception appropriate for HIV positive adolescents with a special focus on hormonal contraceptives. Delaying the start of sexual life and the use of two methods thereafter, one of which is the male condom and the other a highly effective contraceptive method such as hormonal contraception or an intrauterine device, is currently the most effective option for those who desire simultaneous protection from both pregnancy and sexually transmitted diseases. Health care providers should be aware of the possible pharmacokinetic interactions between hormonal contraception and antiretrovirals. There is an urgent need for more information regarding metabolic outcomes of hormonal contraceptives, especially the effect of injectable progestins on bone metabolism, in HIV-positive adolescent girls. PMID- 21631914 TI - Faster Smith-Waterman database searches with inter-sequence SIMD parallelisation. AB - BACKGROUND: The Smith-Waterman algorithm for local sequence alignment is more sensitive than heuristic methods for database searching, but also more time consuming. The fastest approach to parallelisation with SIMD technology has previously been described by Farrar in 2007. The aim of this study was to explore whether further speed could be gained by other approaches to parallelisation. RESULTS: A faster approach and implementation is described and benchmarked. In the new tool SWIPE, residues from sixteen different database sequences are compared in parallel to one query residue. Using a 375 residue query sequence a speed of 106 billion cell updates per second (GCUPS) was achieved on a dual Intel Xeon X5650 six-core processor system, which is over six times more rapid than software based on Farrar's 'striped' approach. SWIPE was about 2.5 times faster when the programs used only a single thread. For shorter queries, the increase in speed was larger. SWIPE was about twice as fast as BLAST when using the BLOSUM50 score matrix, while BLAST was about twice as fast as SWIPE for the BLOSUM62 matrix. The software is designed for 64 bit Linux on processors with SSSE3. Source code is available from http://dna.uio.no/swipe/ under the GNU Affero General Public License. CONCLUSIONS: Efficient parallelisation using SIMD on standard hardware makes it possible to run Smith-Waterman database searches more than six times faster than before. The approach described here could significantly widen the potential application of Smith-Waterman searches. Other applications that require optimal local alignment scores could also benefit from improved performance. PMID- 21631915 TI - Kernel density weighted loess normalization improves the performance of detection within asymmetrical data. AB - BACKGROUND: Normalization of gene expression data has been studied for many years and various strategies have been formulated to deal with various types of data. Most normalization algorithms rely on the assumption that the number of up regulated genes and the number of down-regulated genes are roughly the same. However, the well-known Golden Spike experiment presents a unique situation in which differentially regulated genes are biased toward one direction, thereby challenging the conclusions of previous bench mark studies. RESULTS: This study proposes two novel approaches, KDL and KDQ, based on kernel density estimation to improve upon the basic idea of invariant set selection. The key concept is to provide various importance scores to data points on the MA plot according to their proximity to the cluster of the null genes under the assumption that null genes are more densely distributed than those that are differentially regulated. The comparison is demonstrated in the Golden Spike experiment as well as with simulation data using the ROC curves and compression rates. KDL and KDQ in combination with GCRMA provided the best performance among all approaches. CONCLUSIONS: This study determined that methods based on invariant sets are better able to resolve the problem of asymmetry. Normalization, either before or after expression summary for probesets, improves performance to a similar degree. PMID- 21631916 TI - Multiple Sox genes are expressed in stem cells or in differentiating neuro sensory cells in the hydrozoan Clytia hemisphaerica. AB - BACKGROUND: The Sox genes are important regulators of animal development belonging to the HMG domain-containing class of transcription factors. Studies in bilaterian models have notably highlighted their pivotal role in controlling progression along cell lineages, various Sox family members being involved at one side or the other of the critical balance between self-renewing stem cells/proliferating progenitors, and cells undergoing differentiation. RESULTS: We have investigated the expression of 10 Sox genes in the cnidarian Clytia hemisphaerica. Our phylogenetic analyses allocated most of these Clytia genes to previously-identified Sox groups: SoxB (CheSox2, CheSox3, CheSox10, CheSox13, CheSox14), SoxC (CheSox12), SoxE (CheSox1, CheSox5) and SoxF (CheSox11), one gene (CheSox15) remaining unclassified. In the planula larva and in the medusa, the SoxF orthologue was expressed throughout the endoderm. The other genes were expressed either in stem cells/undifferentiated progenitors, or in differentiating (-ed) cells with a neuro-sensory identity (nematocytes or neurons). In addition, most of them were expressed in the female germline, with their maternal transcripts either localised to the animal region of the egg, or homogeneously distributed. CONCLUSIONS: Comparison with other cnidarians, ctenophores and bilaterians suggest ancient evolutionary conservation of some aspects of gene expression/function at the Sox family level: (i) many Sox genes are expressed in stem cells and/or undifferentiated progenitors; (ii) other genes, or the same under different contexts, are associated with neuro-sensory cell differentiation; (iii) Sox genes are commonly expressed in the germline; (iv) SoxF group genes are associated with endodermal derivatives. Strikingly, total lack of correlation between a given Sox orthology group and expression/function in stem cells/progenitors vs. in differentiating cells implies that Sox genes can easily switch from one side to the other of the balance between these fundamental cellular states in the course of evolution. PMID- 21631917 TI - Dental and prosthodontic status of an over 40 year-old population in Shandong Province, China. AB - BACKGROUND: This study aims to (1) describe the dental status using DMFT for the whole dentition and the anterior, premolar and molar regions; (2) determine associations of demographic variables and socio-economic status (SES) with DMFT and tooth replacement; (3) analyze to what extent the goal as proposed by the WHO -'the retention of not less than 20 teeth throughout life' is achieved. METHODS: DMFT and tooth replacement data of 1588 subjects over 40 years from urban and rural sites in Qingdao (Shandong Province, China) were collected. Relative D, M, and F scores per dental region were calculated and compared by paired T-tests. Multivariable logistic regression was used to determine relationships with age, gender, place of residence, and SES. RESULTS: Mean numbers of D and F were low (1.36 respectively 0.27) at all ages. Molars had highest chance for D and M. For the molar region every additional year of age gave significantly lower chance for D and higher chance for M (OR: 0.98 and 1.02 respectively; both p <= 0.01). Mean number of M was associated with age (approximately 1.5 in each jaw at 40 years and 6 at 80 years). Females had higher chance for D (OR: 1.34; p <= 0.05) and F (OR: 1.69; p <= 0.01), and lower chance for M (OR: 0.60; p <= 0.01). Urban and rural subjects had similar chance for D; urban subjects had approximately 5 times more chance for F (p <= 0.01). SES had no relationship with D and M, however SES low was associated with F (OR: 0.45; p <= 0.01). Replacements were significantly associated with age (all dental regions except anterior region), gender (all dental regions), place of residence (whole dentition and molar region), and SES (whole dentition and premolar and molar regions). CONCLUSIONS: The majority of subjects presented a reduced dentition. Molars were most frequently affected by D and M. D, M, F and replaced teeth were associated with the background variables, however differently for different dental regions. Above the age of 70 years, only 64% of the subjects presented 'not less than 20 natural teeth'. PMID- 21631918 TI - Force of tuberculosis infection among adolescents in a high HIV and TB prevalence community: a cross-sectional observation study. AB - BACKGROUND: Understanding of the transmission dynamics of tuberculosis (TB) in high TB and HIV prevalent settings is required in order to develop effective intervention strategies for TB control. However, there are little data assessing incidence of TB infection in adolescents in these settings. METHODS: We performed a tuberculin skin test (TST) and HIV survey among secondary school learners in a high HIV and TB prevalence community. TST responses to purified protein derivative RT23 were read after 3 days. HIV-infection was assessed using Orasure(r) collection device and ELISA testing. The results of the HIV-uninfected participants were combined with those from previous surveys among primary school learners in the same community, and force of TB infection was calculated by age. RESULTS: The age of 820 secondary school participants ranged from 13 to 22 years. 159 participants had participated in the primary school surveys. At a 10 mm cut off, prevalence of TB infection among HIV-uninfected and first time participants, was 54% (n = 334/620). HIV prevalence was 5% (n = 40/816). HIV infection was not significantly associated with TST positivity (p = 0.07). In the combined survey dataset, TB prevalence was 45% (n = 645/1451), and was associated with increasing age and male gender. Force of infection increased with age, from 3% to 7.3% in adolescents >= 20 years of age. CONCLUSIONS: We show a high force of infection among adolescents, positively associated with increasing age. We postulate this is due to increased social contact with infectious TB cases. Control of the TB epidemic in this setting will require reducing the force of infection. PMID- 21631919 TI - Predicting streptococcal pharyngitis in adults in primary care: a systematic review of the diagnostic accuracy of symptoms and signs and validation of the Centor score. AB - BACKGROUND: Stratifying patients with a sore throat into the probability of having an underlying bacterial or viral cause may be helpful in targeting antibiotic treatment. We sought to assess the diagnostic accuracy of signs and symptoms and validate a clinical prediction rule (CPR), the Centor score, for predicting group A beta-haemolytic streptococcal (GABHS) pharyngitis in adults (> 14 years of age) presenting with sore throat symptoms. METHODS: A systematic literature search was performed up to July 2010. Studies that assessed the diagnostic accuracy of signs and symptoms and/or validated the Centor score were included. For the analysis of the diagnostic accuracy of signs and symptoms and the Centor score, studies were combined using a bivariate random effects model, while for the calibration analysis of the Centor score, a random effects model was used. RESULTS: A total of 21 studies incorporating 4,839 patients were included in the meta-analysis on diagnostic accuracy of signs and symptoms. The results were heterogeneous and suggest that individual signs and symptoms generate only small shifts in post-test probability (range positive likelihood ratio (+LR) 1.45-2.33, -LR 0.54-0.72). As a decision rule for considering antibiotic prescribing (score >= 3), the Centor score has reasonable specificity (0.82, 95% CI 0.72 to 0.88) and a post-test probability of 12% to 40% based on a prior prevalence of 5% to 20%. Pooled calibration shows no significant difference between the numbers of patients predicted and observed to have GABHS pharyngitis across strata of Centor score (0-1 risk ratio (RR) 0.72, 95% CI 0.49 to 1.06; 2-3 RR 0.93, 95% CI 0.73 to 1.17; 4 RR 1.14, 95% CI 0.95 to 1.37). CONCLUSIONS: Individual signs and symptoms are not powerful enough to discriminate GABHS pharyngitis from other types of sore throat. The Centor score is a well calibrated CPR for estimating the probability of GABHS pharyngitis. The Centor score can enhance appropriate prescribing of antibiotics, but should be used with caution in low prevalence settings of GABHS pharyngitis such as primary care. PMID- 21631920 TI - Protein expression dynamics during Escherichia coli glucose-lactose diauxie. AB - BACKGROUND: Escherichia coli is a well-studied anaerobic bacteria which is able to regulate metabolic pathways depending on the type of sugar presented in the medium. We have studied the glucose-lactose shift in E. coli at the protein level using a recently developed mass spectrometry platform. METHOD: Cells were grown in minimal medium containing two sugars (glucose and lactose) and analyzed using novel mass spectrometry cluster. The cluster combines the high resolving power and dynamic range of Fourier transform ion cyclotron resonance (FTICR) for accurate mass measurement and quantitation with multiple ion traps for fast and sensitive tandem mass spectrometry. The protein expression profile was followed in time across the glucose-lactose diauxic shift using label-free quantitation from the FTICR data. RESULTS AND CONCLUSION: The entire dataset was interrogated by KEGG pathway analysis, mapping measured changes in protein abundance onto known metabolic pathways. The obtained results were consistent with previously published gene expression data, with beta-galactosidase being the most strongly induced protein during the diauxic shift. PMID- 21631921 TI - Childhood and adolescent predictors of leisure time physical activity during the transition from adolescence to adulthood: a population based cohort study. AB - BACKGROUND: Few studies have investigated factors that influence physical activity behavior during the transition from adolescence to adulthood. This study explores the associations of sociodemographic, behavioral, sociocultural, attitudinal and physical factors measured in childhood and adolescence with physical activity behavior during the transition from adolescence to adulthood. METHODS: Childhood and adolescent data (at ages 7-15 years) were collected as part of the 1985 Australian Health and Fitness Survey and subdivided into sociodemographics (socioeconomic status, parental education), behavioral (smoking, alcohol, sports diversity, outside school sports), sociocultural (active father, active mother, any older siblings, any younger siblings, language spoken at home), attitudinal (sports/recreational competency, self-rated health, enjoyment physical education/physical activity, not enjoying school sports) and physical (BMI, time taken to run 1.6 km, long jump) factors. Physical activity between the ages 15 and 29 years was reported retrospectively using the Historical Leisure Activity Questionnaire at follow-up in 2004-2006 by 2,048 participants in the Childhood Determinants of Adult Health Study (CDAH). Australia's physical activity recommendations for children and adults were used to categorize participants as persistently active, variably active or persistently inactive during the transition from adolescence to adulthood. RESULTS: For females, perceived sports competency in childhood and adolescence was significantly associated with being persistently active (RR=1.88, 95% CI=1.39, 2.55). Smoking (RR=0.31 CI=0.12, 0.82) and having younger siblings (RR=0.69 CI=0.52, 0.93) were inversely associated with being persistently active after taking physical and attitudinal factors into account. For males, playing sport outside school (RR=1.47 CI=1.05, 2.08), having active fathers (RR=1.25 CI=1.01, 1.54) and not enjoying school sport (RR=4.07 CI=2.31, 7.17) were associated with being persistently active into adulthood. Time taken to complete the 1.6 km run was inversely associated with being persistently active into adulthood (RR=0.85 CI=0.78, 0.93) after adjusting for recreational competency. CONCLUSIONS: Perceived sports competency (females) and cardiorespiratory fitness, playing sport outside school and having active fathers (males) in childhood and adolescence were positively associated with being persistently active during the transition from adolescence to adulthood. PMID- 21631922 TI - Immunohistochemical localization of mu opioid receptor in the marginal division with comparison to patches in the neostriatum of the rat brain. AB - BACKGROUND: Mu opioid receptor (MOR), which plays key roles in analgesia and also has effects on learning and memory, was reported to distribute abundantly in the patches of the neostriatum. The marginal division (MrD) of the neostriatum, which located at the caudomedial border of the neostriatum, was found to stain for enkephalin and substance P immunoreactivities and this region was found to be involved in learning and memory in our previous study. However, whether MOR also exists in the MrD has not yet been determined. METHODS: In this study, we used western blot analysis and immunoperoxidase histochemical methods with glucose oxidase-DAB-nickel staining to investigate the expression of MOR in the MrD by comparison to the patches in the neostriatum. RESULTS: The results from western blot analyses revealed that the antibody to MOR detected a 53 kDa protein band, which corresponded directly to the molecular weight of MOR. Immunohistochemical results showed that punctate MOR-immunoreacted fibers were observed in the "patch" areas in the rostrodorsal part of the neostriatum but these previous studies showed neither labelled neuronal cell bodies, nor were they shown in the caudal part of the neostriatum. Dorsoventrally oriented dark MOR-immunoreactive nerve fibers with individual labelled fusiform cell bodies were firstly observed in the band at the caudomedial border, the MrD, of the neostriatum. The location of the MOR-immunoreactivity was in the caudomedial border of the neostriatum. The morphology of the labelled fusiform neuronal somatas and the dorsoventrally oriented MOR-immunoreacted fibers in the MrD was distinct from the punctate MOR immunoreactive diffuse mosaic-patterned patches in the neostriatum. CONCLUSIONS: The results indicated that MOR was expressed in the MrD as well as in patches in the neostriatum of the rat brain, but with different morphological characteristics. The punctate MOR-immunoreactive and diffuse mosaic-patterned patches were located in the rostrodorsal part of the neostriatum. By contrast, in the MrD, the dorsoventrally parallel oriented MOR-immunoreactive fibers with individual labelled fusiform neuronal somatas were densely packed in the caudomedial border of the neostriatum. The morphological difference in MOR immunoreactivity between the MrD and the patches indicated potential functional differences between them. The MOR most likely plays a role in learning and memory associated functions of the MrD. PMID- 21631923 TI - Does pre-operative psychological distress affect patient satisfaction after primary total hip arthroplasty? AB - BACKGROUND: There are concerns that pre-operative psychological distress might be associated with reduced patient satisfaction after total hip replacement (THR). METHODS: We investigated this in a multi-centre prospective study between January 1999 and January 2002. We dichotomised the patients into the mentally distressed (MHS <= 56) and the not mentally distressed (MHS > 56) groups based on their pre operative Mental Health Score (MHS) of SF36. RESULTS: 448 patients (340 not distressed and 108 distressed) completed the patient satisfaction survey. Patient satisfaction rate at five year was 96.66% (415/448). There was no difference in patient satisfaction or willingness to have the surgery between the two groups. None of pre-operative variables predicted five year patient satisfaction in logistic regression. CONCLUSIONS: Patient satisfaction after surgery may not be adversely affected by pre-operative psychological distress. PMID- 21631924 TI - Plasma levels of leptin and soluble leptin receptor and polymorphisms of leptin gene -18G > A and leptin receptor genes K109R and Q223R, in survivors of childhood acute lymphoblastic leukemia. AB - BACKGROUND: Approximately 20% of children and adolescents in Europe are overweight. Survivors of pediatric acute lymphoblastic leukemia (ALL) are at increased risk of overweight and obesity. The purpose of this study was to assess leptin and leptin soluble receptor levels, as well as polymorphisms of selected genes in survivors of pediatric ALL, and the influence of chemo- and radiotherapy on development of overweight in the context of leptin regulation. METHODS: Eighty two patients (55% males), of median age 13.2 years (m: 4.8 years; M: 26.2 years) were included in the study. The ALL therapy was conducted according to modified Berlin-Frankfurt-Munster (BFM; n = 69) regimen or New York (n = 13) regimen. In 38% of patients cranial radiotherapy (CRT) was used in median dose of 18.2Gy (m: 14Gy; M: 24Gy). Median age at diagnosis was 4.5 (m: 1 year; M: 16.9 years) and median time from completion of ALL treatment was 3.2 years (m: 0.5 year; M: 4.3 years). Patients with BMI >=85 percentile were classified as overweight. Correlation of plasma levels of leptin and leptin soluble receptor, and polymorphisms of leptin gene -18G > A, leptin receptor genes K109R and Q223R, and the overweight status were analyzed in relation to gender, intensity of chemotherapy (high intensity vs. standard intensity regimens) and to the use of CRT. RESULTS: Significant differences of leptin levels in patients treated with and without CRT, both in the entire study group (22.2+/- 3.13 ng/ml vs. 14.9+/ 1.6 ng/ml; p < 0.03) and in female patients (29.9+/-4.86 ng/ml vs. 16.9+/-2.44 ng/ml; p = 0.014), were found. Significant increase of leptin levels was also found in overweight patients compared to the non-overweight patients in the entire study group (29.2+/-2.86 ng/ml vs. 12.6+/-1.51 ng/ml; p < 0.0001), female patients (35.4+/-6.48 ng/ml vs. 18.4+/-2.5 ng/ml; p = 0.005), and male patients (25.7+/-2.37 ng/ml vs. 6.9+/-0.95 ng/ml; p < 0.0001). Negative correlation was observed for plasma levels of soluble leptin receptor and overweight status, with significant differences in overweight and non-overweight patients, both in the entire study group (18.2+/-0.75 ng/ml vs. 20.98+/-0.67 ng/ml; p = 0.017) and in male patients (18.2+/-1.03 ng/ml vs. 21.8+/- 1.11 ng/ml; p = 0.038). Significant (p < 0.05) negative correlation was found between leptin and leptin receptor levels in the entire group (correlation coefficient: 0.393) and in both gender subgroups (correlation coefficient in female patients: -0.427; in male patients: 0.396). CONCLUSIONS: The prevalence of overweight in our cohort was higher than in general European population (31% vs 20%) and increased regardless of the use of CRT. Leptin and leptin receptor levels may be used as useful markers of high risk of becoming overweight in ALL survivors, particularly in females treated with CRT. Polymorphisms of leptin gene -18G > A and leptin receptor genes K109R and Q223R were not associated with overweight status in ALL survivors. PMID- 21631925 TI - A study on the immunological basis of the dissociation between type I hypersensitivity skin reactions to Blomia tropicalis antigens and serum anti-B. tropicalis IgE antibodies. AB - BACKGROUND: Two conditions are used as markers of atopy: the presence of circulating anti-allergen IgE antibodies and the presence of positive skin prick test (SPT) reactions to allergenic extracts. The correlation between these conditions is not absolute. This study aimed at investigating immunological parameters that may mediate this lack of correlation. Individuals whose sera contained anti-B. tropicalis extract IgE antibodies (alpha-BtE IgE) were divided into two groups, according to the presence or absence of skin reactivity to B. tropicalis extract (BtE). The following parameters were investigated: total IgE levels; alpha-BtE IgE levels; an arbitrary alpha-BtE IgE/total IgE ratio; the proportion of carbohydrate-reactive alpha-BtE IgE; the proportion of alpha-BtE IgE that reacted with Ascaris lumbricoides extract (AlE); the production of IL-10 by BtE- and AlE-stimulated peripheral blood cells (PBMC). RESULTS: Total IgE levels were similar in the two groups, but alpha-BtE IgE was significantly higher in the SPT-positive group (SPT+). A large overlap of alpha-BtE IgE levels was found in individuals of both groups, indicating that these levels alone cannot account for the differences in SPT outcome. Individuals of the two groups did not differ, statistically, in the proportion of alpha-BtE IgE that reacted with carbohydrate and in the production of IL-10 by BtE- and AlE-stimulated PBMC. Both groups had part of alpha-BtE IgE activity absorbed out by AlE, indicating the existence of cross-reactive IgE antibodies. However, the alpha-BtE IgE from the SPT-negative individuals (SPT-) was more absorbed with AlE than the alpha-BtE IgE from the SPT+ individuals. This finding may be ascribed to avidity differences of the alpha-BtE IgE that is present in the two groups of individuals, and could occur if at least part of the alpha-BtE IgE from the SPT- individuals were elicited by A. lumbricoides infection. CONCLUSION: The present results suggest that a low ratio of specific IgE to total IgE levels (in a minority of individuals), and differences in alpha-BtE IgE avidities (which would have high affinities for A. lumbricoides antigens in SPT- than in SPT+ individuals) may play a role in the down-modulation of type-I hypersensitivity reaction against aeroallergens described in helminth-infected individuals. PMID- 21631926 TI - Genes and queens. PMID- 21631927 TI - The Chinese-version of the CARE measure reliably differentiates between doctors in primary care: a cross-sectional study in Hong Kong. AB - BACKGROUND: The Consultation and Relational Empathy (CARE) Measure is a widely used patient-rated experience measure which has recently been translated into Chinese and has undergone preliminary qualitative and quantitative validation. The objective of this study was to determine the reliability of the Chinese version of the CARE Measure in reliably differentiating between doctors in a primary care setting in Hong Kong METHODS: Data were collected from 984 primary care patients attending 20 doctors with differing levels of training in family medicine in 5 public clinics in Hong Kong. The acceptability of the Chinese-CARE measure to patients was assessed. The reliability of the measure in discriminating effectively between doctors was analysed by Generalisability theory (G-Theory) RESULTS: The items in the Chinese-CARE measure were regarded as important by patients and there were few 'not applicable' responses. The measure showed high internal reliability (coefficient 0.95) and effectively differentiated between doctors with only 15-20 patient ratings per doctor (inter rater reliability > 0.8). Doctors' mean CARE measure scores varied widely, ranging from 24.1 to 45.9 (maximum possible score 50) with a mean of 34.6. CARE Measure scores were positively correlated with level of training in family medicine (Spearman's rho 0.493, p < 0.05). CONCLUSION: These data demonstrate the acceptability, feasibility and reliability of using the Chinese-CARE Measure in primary care in Hong Kong to differentiate between doctors interpersonal competencies. Training in family medicine appears to enhance these key interpersonal skills. PMID- 21631928 TI - Rapid assessment response (RAR) study: drug use and health risk - Pretoria, South Africa. AB - BACKGROUND: Within a ten year period South Africa has developed a substantial illicit drug market. Data on HIV risk among drug using populations clearly indicate high levels of HIV risk behaviour due to the sharing of injecting equipment and/or drug-related unprotected sex. While there is international evidence on and experience with adequate responses, limited responses addressing drug use and drug-use-related HIV and other health risks are witnessed in South Africa. This study aimed to explore the emerging problem of drug-related HIV transmission and to stimulate the development of adequate health services for the drug users, by linking international expertise and local research. METHODS: A Rapid Assessment and Response (RAR) methodology was adopted for the study. For individual and focus group interviews a semi-structured questionnaire was utilised that addressed key issues. Interviews were conducted with a total of 84 key informant (KI) participants, 63 drug user KI participants (49 males, 14 females) and 21 KI service providers (8 male, 13 female). RESULTS AND DISCUSSION: Adverse living conditions and poor education levels were cited as making access to treatment harder, especially for those living in disadvantaged areas. Heroin was found to be the substance most available and used in a problematic way within the Pretoria area. Participants were not fully aware of the concrete health risks involved in drug use, and the vague ideas held appear not to allow for concrete measures to protect themselves. Knowledge with regards to substance related HIV/AIDS transmission is not yet widespread, with some information sources disseminating incorrect or unspecific information. CONCLUSIONS: The implementation of pragmatic harm-reduction and other evidence-based public health care policies that are designed to reduce the harmful consequences associated with substance use and HIV/AIDS should be considered. HIV testing and treatment services also need to be made available in places accessed by drug users. PMID- 21631929 TI - No relevant cardiac, pharmacokinetic or safety interactions between roflumilast and inhaled formoterol in healthy subjects: an open-label, randomised, actively controlled study. AB - BACKGROUND: Roflumilast is an oral, selective phosphodiesterase 4 inhibitor with anti-inflammatory effects in chronic obstructive pulmonary disease (COPD). The addition of roflumilast to long-acting bronchodilators improves lung function in patients with moderate-to-severe COPD. The present study investigated drug-drug interaction effects between inhaled formoterol and oral roflumilast. METHODS: This was a single-centre (investigational clinic), open, randomised, multiple dose, parallel-group study. In Regimen A, healthy men were treated with roflumilast (500 MUg tablet once daily; Day 2-18) and concomitant formoterol (24 MUg twice daily; Day 12-18). In Regimen B, healthy men were treated with formoterol (24 MUg twice daily; Day 2-18) and concomitant roflumilast (500 MUg once daily; Day 9-18). Steady-state plasma pharmacokinetics of roflumilast, roflumilast N-oxide and/or formoterol (Cmax and AUC0-tau) as well as pharmacodynamics - blood pressure, transthoracic impedance cardiography (ZCG), 12 lead digital electrocardiography, peripheral blood eosinophils, and serum glucose and potassium concentrations - were evaluated through Day 1 (baseline), Day 8 (Regimen B: formoterol alone) or Day 11 (Regimen A: roflumilast alone), and Day 18 (Regimen A and B: roflumilast plus formoterol). Blood and urine samples were taken for safety assessment at screening, pharmacokinetic profiling days and Day 19. Adverse events were monitored throughout the study. RESULTS: Of the 27 subjects enrolled, 24 were evaluable (12 in each regimen). No relevant pharmacokinetic interactions occurred. Neither roflumilast nor formoterol were associated with significant changes in cardiovascular parameters as measured by ZCG, and these parameters were not affected during concomitant administration. Formoterol was associated with a slight increase in heart rate and a corresponding shortening of the QT interval, without changes in the heart rate corrected QTc interval. There were small effects on the other pharmacodynamic assessments when roflumilast and formoterol were administered individually, but no interactions or safety concerns were seen after concomitant administration. No severe or serious adverse events were reported, and no adverse events led to premature study discontinuation. CONCLUSIONS: No clinically relevant pharmacokinetic or pharmacodynamic interactions were found when oral roflumilast was administered concomitantly with inhaled formoterol, including no effect on cardiac repolarisation. Roflumilast was well tolerated. PMID- 21631930 TI - Non-specific physical symptoms in relation to actual and perceived proximity to mobile phone base stations and powerlines. AB - BACKGROUND: Evidence about a possible causal relationship between non-specific physical symptoms (NSPS) and exposure to electromagnetic fields (EMF) emitted by sources such as mobile phone base stations (BS) and powerlines is insufficient. So far little epidemiological research has been published on the contribution of psychological components to the occurrence of EMF-related NSPS. The prior objective of the current study is to explore the relative importance of actual and perceived proximity to base stations and psychological components as determinants of NSPS, adjusting for demographic, residency and area characteristics. METHODS: Analysis was performed on data obtained in a cross sectional study on environment and health in 2006 in the Netherlands. In the current study, 3611 adult respondents (response rate: 37%) in twenty-two Dutch residential areas completed a questionnaire. Self-reported instruments included a symptom checklist and assessment of environmental and psychological characteristics. The computation of the distance between household addresses and location of base stations and powerlines was based on geo-coding. Multilevel regression models were used to test the hypotheses regarding the determinants related to the occurrence of NSPS. RESULTS: After adjustment for demographic and residential characteristics, analyses yielded a number of statistically significant associations: Increased report of NSPS was predominantly predicted by higher levels of self-reported environmental sensitivity; perceived proximity to base stations and powerlines, lower perceived control and increased avoidance (coping) behavior were also associated with NSPS. A trend towards a moderator effect of perceived environmental sensitivity on the relation between perceived proximity to BS and NSPS was verified (p = 0.055). There was no significant association between symptom occurrence and actual distance to BS or powerlines. CONCLUSIONS: Perceived proximity to BS, psychological components and socio demographic characteristics are associated with the report of symptomatology. Actual distance to the EMF source did not show up as determinant of NSPS. PMID- 21631931 TI - Survival and associated factors in 268 adults with Pompe disease prior to treatment with enzyme replacement therapy. AB - BACKGROUND: Pompe disease is a rare lysosomal storage disorder characterized by muscle weakness and wasting. The majority of adult patients have slowly progressive disease, which gradually impairs mobility and respiratory function and may lead to wheelchair and ventilator dependency. It is as yet unknown to what extent the disease reduces the life span of these patients. Our objective was to determine the survival of adults with Pompe disease not receiving ERT and to identify prognostic factors associated with survival. METHODS: Data of 268 patients were collected in a prospective international observational study conducted between 2002 and 2009. Survival analyses from time of diagnosis and from time of study entry were performed using Kaplan-Meier curves and Cox proportional-hazards regression. RESULTS: Median age at study entry was 48 years (range 19-79 years). Median survival after diagnosis was 27 years, while median age at diagnosis was 38 years. During follow-up, twenty-three patients died prior to ERT, with a median age at death of 55 (range 23-77 years). Use of wheelchair and/or respiratory support and patients' score on the Rotterdam Handicap Scale (RHS) were identified as prognostic factors for survival. Five-year survival for patients without a wheelchair or respiratory support was 95% compared to 74% in patients who were wheelchair-bound and used respiratory support. In a Dutch subgroup of 99 patients, we compared the observed number of deaths to the expected number of deaths in the age- and sex-matched general population. During a median follow-up of 2.3 years, the number of deaths among the Dutch Pompe patients was higher than the expected number of deaths in the general population. CONCLUSION: Our study shows for the first time that untreated adults with Pompe disease have a higher mortality than the general population and that their levels of disability and handicap/participation are the most important factors associated with mortality. These results may be of relevance when addressing the effect of ERT or other potential treatment options on survival. PMID- 21631932 TI - Paracetamol versus placebo in treatment of non-severe malaria in children in Guinea-Bissau: a randomized controlled trial. AB - BACKGROUND: The current guidelines for treatment of malaria include paracetamol to children with fever. No convincing evidence for the beneficial effects of this practice exists. Studies show that time to parasite clearance is significantly longer in children treated with paracetamol, which questions the policy. Whether this is of clinical importance has not been investigated. METHODS: Children with Plasmodium falciparum monoinfection and >=20 parasites per 200 leucocytes at the Bandim Health Centre, Guinea-Bissau were randomized to receive paracetamol or placebo together with chloroquine for three days in a double blind randomized study. Temperature and symptoms were recorded twice daily during treatment and on day 3. The participants were interviewed and a malaria film taken once weekly until day 35. The data is in the form of grouped failure-times, the outcome of interest being time until parasitaemia during follow-up. Mantel-Haenszel weighted odds ratios are given. Other differences between and within the two groups have been tested using the Chi-square test and Mann-Whitney U test. RESULTS: In the evening of the day of inclusion, the temperature was slightly, but statistically insignificant, higher in the placebo group and significantly more children complained of headache. At no other time was a significant difference in temperature or symptoms detected. However, 6 children from the placebo-group as compared to two children from the paracetamol-group were admitted to hospital with high fever and convulsions by day 3. No differences in the cumulative percentages of children with adequate clinical and parasitological response were found in the intention-to-treat analysis or in the per-protocol analysis. CONCLUSION: Fewer children had early treatment failure and the mean temperature was slightly lower in the afternoon on day 0 in the paracetamol group. However, the cumulative adequate clinical and parasitological cure rates were not significantly different during the period of study. It is doubtful whether adding paracetamol to the treatment of uncomplicated malaria in children is beneficial. TRIAL REGISTRATION: NCT00137566. PMID- 21631933 TI - Exploring dietitians' salient beliefs about shared decision-making behaviors. AB - BACKGROUND: Shared decision making (SDM), a process by which health professionals and patients go through the decision-making process together to agree on treatment, is a promising strategy for promoting diet-related decisions that are informed and value based and to which patients adhere well. The objective of the present study was to identify dietitians' salient beliefs regarding their exercise of two behaviors during the clinical encounter, both of which have been deemed essential for SDM to take place: (1) presenting patients with all dietary treatment options for a given health condition and (2) helping patients clarify their values and preferences regarding the options. METHODS: Twenty-one dietitians were allocated to four focus groups. Facilitators conducted the focus groups using a semistructured interview guide based on the Theory of Planned Behavior. Discussions were audiotaped, transcribed verbatim, coded, and analyzed with NVivo8 (QSR International, Cambridge, MA) software. RESULTS: Most participants stated that better patient adherence to treatment was an advantage of adopting the two SDM behaviors. Dietitians identified patients, physicians, and the multidisciplinary team as normative referents who would approve or disapprove of their adoption of the SDM behaviors. The most often reported barriers and facilitators for the behaviors concerned patients' characteristics, patients' clinical situation, and time. CONCLUSIONS: The implementation of SDM in nutrition clinical practice can be guided by addressing dietitians' salient beliefs. Identifying these beliefs also provides the theoretical framework needed for developing a quantitative survey questionnaire to further study the determinants of dietitians' adoption of SDM behaviors. PMID- 21631934 TI - Establishment of real-time quantitative reverse transcription polymerase chain reaction assay for transcriptional analysis of duck enteritis virus UL55 gene. AB - BACKGROUND: Real-time quantitative reverse transcription polymerase chain reaction assay (qRT-PCR) has become the benchmark for detection and quantification of target gene expression level and been utilized increasingly in detection of viral load and therapy monitoring. The dynamic transcription variation of duck enteritis virus UL55 gene during the life cycle of duck enteritis virus in infected cells has not been reported yet. RESULTS: The newly identified duck enteritis virus UL55 gene was amplified and cloned into pMD18-T vector after digestion to generate a recombinant plasmid pMD18-T/UL55 for the establishment of qRT-PCR as standard DNA. The results of agarose gel electrophoresis and melting curve analysis demonstrated the primers we designed for qRT-PCR were specific and available. We used beta-actin as a reference gene for normalization and established two standard curves based on pMD18-T/UL55 and pMD18-T/beta-actin successfully. Based on that, the transcriptional analysis of DEV UL55 gene was performed, and the result suggested the expression of UL55 mRNA was at a low level from 0 to 8 h post-infection(p.i.), then accumulated quickly since 12 h p.i. and peaked at 36 h p.i., it can be detected till 60 h p.i.. Nucleic acid inhibition test was carried out for analyzing a temporal regulation condition of DEV UL55 gene, result revealed that it was sensitive to ganciclovir. Synthesis procedures of DEV UL55 gene can be inhibited by ganciclovir. CONCLUSIONS: The method we established in this paper can provide quantitative values reflecting the amounts of measured mRNA in samples. It's available for detection and quantification, also can be used in DEV diagnosis. The DEV UL55 gene was produced most abundantly during the late phase of replication in DEV infected cells and the transcription of it depended on the synthesized DNA. DEV UL55 gene is a gamma2 gene which occurs last and have a strict requirement for viral DNA synthesis. PMID- 21631935 TI - Risky sexual practices and related factors among ART attendees in Addis Ababa Public Hospitals, Ethiopia: a cross-sectional study. AB - BACKGROUND: Many HIV-positive persons avoid risky sexual practices after testing HIV sero-positive. However, a substantial number continue to engage in risky sexual practices that may further transmit the virus, put them at risk of contracting secondary sexually transmitted infections and lead to problems with drug resistance. Thus, this study was intended to assess risky sexual practices and related factors among HIV- positive ART attendees in public hospitals of Addis Ababa. METHODS: A cross-sectional study was conducted among ART attendees from February to March, 2009. Questionnaire-based face-to-face interviews were used to gather data. SPSS software was used to perform descriptive and logistic regression analyses. RESULTS: Six hundred and one ART attendees who fulfilled the inclusion criteria was included in the study and interviewed. More than one-third (36.9%) had a history of risky sexual practices in the three months prior to the study. The major reasons given for not using condoms were: partner's dislike of them, both partners being positive for HIV and the desire to have a child. Factors associated with risky sexual practices included: lack of discussion about condom use (Adjusted Odds Ratio (AOR = 7.23, 95% CI: 4.14, 12.63); lack of self efficacy in using condoms (AOR = 3.29, 95% CI: 2.07, 5.23); lack of sexual pleasure when using a condom (AOR = 2.39, 95% CI: 1.52, 3.76); and multiple sexual partners (AOR = 2.67, 95% CI: 1.09, 6.57). Being with a negative sero status partner (AOR = 0.33, 95% CI: 0.14, 0.80), or partners of unknown sero status (AOR = 0.19, 95% CI: 0.09, 0.39) were associated with less risky practice. CONCLUSIONS: A considerable proportion (36.9%) of respondents engaged in unprotected sexual intercourse, potentially resulting in re-infection by a new virus strain, other sexually transmitted infections and onward transmission of the HIV virus. Health education and counseling which focuses on the identified factors has to be provided. The health education and counseling can be provided to these people at ART appointments on follow- up care. It can be provided in a one-on-one basis or through patient group educational discussions at the clinics. PMID- 21631936 TI - Communication, advice exchange and job satisfaction of nursing staff: a social network analyses of 35 long-term care units. AB - BACKGROUND: The behaviour of individuals is affected by the social networks in which they are embedded. Networks are also important for the diffusion of information and the influence of employees in organisations. Yet, at the moment little is known about the social networks of nursing staff in healthcare settings. This is the first study that investigates informal communication and advice networks of nursing staff in long-term care. We examine the structure of the networks, how they are related to the size of units and characteristics of nursing staff, and their relationship with job satisfaction. METHODS: We collected social network data of 380 nursing staff of 35 units in group projects and psychogeriatric units in nursing homes and residential homes in the Netherlands. Communication and advice networks were analyzed in a social network application (UCINET), focusing on the number of contacts (density) between nursing staff on the units. We then studied the correlation between the density of networks, size of the units and characteristics of nursing staff. We used multilevel analyses to investigate the relationship between social networks and job satisfaction of nursing staff, taking characteristics of units and nursing staff into account. RESULTS: Both communication and advice networks were negatively related to the number of residents and the number of nursing staff of the units. Communication and advice networks were more dense when more staff worked part-time. Furthermore, density of communication networks was positively related to the age of nursing staff of the units. Multilevel analyses showed that job satisfaction differed significantly between individual staff members and units and was influenced by the number of nursing staff of the units. However, this relationship disappeared when density of communication networks was added to the model. CONCLUSIONS: Overall, communication and advice networks of nursing staff in long-term care are relatively dense. This fits with the high level of cooperation that is needed to provide good care to residents. Social networks are more dense in small units and are also shaped by characteristics of staff members. The results furthermore show that communication networks are important for staff's job satisfaction. PMID- 21631937 TI - Distribution of silver in rats following 28 days of repeated oral exposure to silver nanoparticles or silver acetate. AB - BACKGROUND: The study investigated the distribution of silver after 28 days repeated oral administration of silver nanoparticles (AgNPs) and silver acetate (AgAc) to rats. Oral administration is a relevant route of exposure because of the use of silver nanoparticles in products related to food and food contact materials. RESULTS: AgNPs were synthesized with a size distribution of 14 +/- 4 nm in diameter (90% of the nanoparticle volume) and stabilized in aqueous suspension by the polymer polyvinylpyrrolidone (PVP). The AgNPs remained stable throughout the duration of the 28-day oral toxicity study in rats. The organ distribution pattern of silver following administration of AgNPs and AgAc was similar. However the absolute silver concentrations in tissues were lower following oral exposure to AgNPs. This was in agreement with an indication of a higher fecal excretion following administration of AgNPs. Besides the intestinal system, the largest silver concentrations were detected in the liver and kidneys. Silver was also found in the lungs and brain. Autometallographic (AMG) staining revealed a similar cellular localization of silver in ileum, liver, and kidney tissue in rats exposed to AgNPs or AgAc. Using transmission electron microscopy (TEM), nanosized granules were detected in the ileum of animals exposed to AgNPs or AgAc and were mainly located in the basal lamina of the ileal epithelium and in lysosomes of macrophages within the lamina propria. Using energy dispersive x ray spectroscopy it was shown that the granules in lysosomes consisted of silver, selenium, and sulfur for both AgNP and AgAc exposed rats. The diameter of the deposited granules was in the same size range as that of the administered AgNPs. No silver granules were detected by TEM in the liver. CONCLUSIONS: The results of the present study demonstrate that the organ distribution of silver was similar when AgNPs or AgAc were administered orally to rats. The presence of silver granules containing selenium and sulfur in the intestinal wall of rats exposed to either of the silver forms suggests a common mechanism of their formation. Additional studies however, are needed to gain further insight into the underlying mechanisms of the granule formation, and to clarify whether AgNPs dissolve in the gastrointestinal system and/or become absorbed and translocate as intact nanoparticles to organs and tissues. PMID- 21631938 TI - Plasma proteome changes in cardiovascular disease patients: novel isoforms of apolipoprotein A1. AB - BACKGROUND: The aim of this proteomic study was to look for changes taking place in plasma proteomes of patients with acute myocardial infarction (AMI), unstable angina pectoris (UAP), and stable angina pectoris (SAP). METHODS: Depleted plasma proteins were separated by 2D SDS-PAGE (pI 4-7), and proteomes were compared using Progenesis SameSpots statistical software. Proteins were identified by nanoLC-MS/MS. Proteins were quantified using commercial kits. Apolipoprotein A1 was studied using 1D and 2D SDS-PAGE, together with western blotting. RESULTS: Reciprocal comparison revealed 46 unique, significantly different spots; proteins in 34 spots were successfully identified and corresponded to 38 different proteins. Discrete comparisons of patient groups showed 45, 41, and 8 significantly different spots when AMI, UAP, and SAP were compared with the control group. On the basis of our proteomic data, plasma levels of two of them, alpha-1 microglobulin and vitamin D-binding protein, were determined. The data, however, failed to prove the proteins to be suitable markers or risk factors in the studied groups. The plasma level and isoform representation of apolipoprotein A1 were also estimated. Using 1D and 2D SDS-PAGE, together with western blotting, we observed extra high-molecular weight apolipoprotein A1 fractions presented only in the patient groups, indicating that the novel high-molecular weight isoforms of apolipoprotein A1 may be potential new markers or possible risk factors of cardiovascular disease. CONCLUSION: The reported data show plasma proteome changes in patients with AMI, UAP, and SAP. We propose some apolipoprotein A1 fractions as a possible new disease-associated marker of cardiovascular disorders. PMID- 21631939 TI - Polyomic profiling reveals significant hepatic metabolic alterations in glucagon receptor (GCGR) knockout mice: implications on anti-glucagon therapies for diabetes. AB - BACKGROUND: Glucagon is an important hormone in the regulation of glucose homeostasis, particularly in the maintenance of euglycemia and prevention of hypoglycemia. In type 2 Diabetes Mellitus (T2DM), glucagon levels are elevated in both the fasted and postprandial states, which contributes to inappropriate hyperglycemia through excessive hepatic glucose production. Efforts to discover and evaluate glucagon receptor antagonists for the treatment of T2DM have been ongoing for approximately two decades, with the challenge being to identify an agent with appropriate pharmaceutical properties and efficacy relative to potential side effects. We sought to determine the hepatic & systemic consequence of full glucagon receptor antagonism through the study of the glucagon receptor knock-out mouse (Gcgr-/-) compared to wild-type littermates. RESULTS: Liver transcriptomics was performed using Affymetric expression array profiling, and liver proteomics was performed by iTRAQ global protein analysis. To complement the transcriptomic and proteomic analyses, we also conducted metabolite profiling (~200 analytes) using mass spectrometry in plasma. Overall, there was excellent concordance (R = 0.88) for changes associated with receptor knock-out between the transcript and protein analysis. Pathway analysis tools were used to map the metabolic processes in liver altered by glucagon receptor ablation, the most notable being significant down-regulation of gluconeogenesis, amino acid catabolism, and fatty acid oxidation processes, with significant up-regulation of glycolysis, fatty acid synthesis, and cholesterol biosynthetic processes. These changes at the level of the liver were manifested through an altered plasma metabolite profile in the receptor knock-out mice, e.g. decreased glucose and glucose-derived metabolites, and increased amino acids, cholesterol, and bile acid levels. CONCLUSIONS: In sum, the results of this study suggest that the complete ablation of hepatic glucagon receptor function results in major metabolic alterations in the liver, which, while promoting improved glycemic control, may be associated with adverse lipid changes. PMID- 21631940 TI - Analytical methods applied to diverse types of Brazilian propolis. AB - Propolis is a bee product, composed mainly of plant resins and beeswax, therefore its chemical composition varies due to the geographic and plant origins of these resins, as well as the species of bee. Brazil is an important supplier of propolis on the world market and, although green colored propolis from the southeast is the most known and studied, several other types of propolis from Apis mellifera and native stingless bees (also called cerumen) can be found. Propolis is usually consumed as an extract, so the type of solvent and extractive procedures employed further affect its composition. Methods used for the extraction; analysis the percentage of resins, wax and insoluble material in crude propolis; determination of phenolic, flavonoid, amino acid and heavy metal contents are reviewed herein. Different chromatographic methods applied to the separation, identification and quantification of Brazilian propolis components and their relative strengths are discussed; as well as direct insertion mass spectrometry fingerprinting.Propolis has been used as a popular remedy for several centuries for a wide array of ailments. Its antimicrobial properties, present in propolis from different origins, have been extensively studied. But, more recently, anti-parasitic, anti-viral/immune stimulating, healing, anti tumor, anti-inflammatory, antioxidant and analgesic activities of diverse types of Brazilian propolis have been evaluated. The most common methods employed and overviews of their relative results are presented. PMID- 21631941 TI - MiR-204 regulates cardiomyocyte autophagy induced by ischemia-reperfusion through LC3-II. AB - BACKGROUND: Autophagy plays a significant role in myocardial ischemia-reperfusion (IR) injury. So it is important to inhibit autophagy to protect cardiomyocytes besides anti-apoptosis. MiRNA has been demonstrated to protect cardiomyocytes against apoptosis during IR, while whether it has anti-autophagy effect has not been known. The aim of this study was to investigate whether miR-204 regulated autophagy by regulating LC3-II protein, which is the marker of autophagosome during myocardial IR injury. METHODS: Adult SD rats were randomized to Control and IR groups. IR group was treated with 30 min ischemia by ligating the left anterior descending coronary artery, followed by 2 h reperfusion by loosing the ligation. The expression of miR-204 was measured by RT-PCR, and LC3 protein was measured by western-blot. RESULTS: We found that IR induced cardiomyocytes autophagy, together with down-regulation of miR-204 and up-regulation of LC3-II protein. And, we have found that LC3-II protein was regulated by miR-204, using the method of transferring miR-204 mimic or AMO-204 into the cardiomyocytes, before. CONCLUSIONS: These studies provided evidence that miR-204 played an important role in regulating autophagy through LC3-IIprotein during IR. PMID- 21631942 TI - Reduction of mutant huntingtin accumulation and toxicity by lysosomal cathepsins D and B in neurons. AB - BACKGROUND: Huntington's disease is caused by aggregation of mutant huntingtin (mHtt) protein containing more than a 36 polyQ repeat. Upregulation of macroautophagy was suggested as a neuroprotective strategy to degrade mutant huntingtin. However, macroautophagy initiation has been shown to be highly efficient in neurons whereas lysosomal activities are rate limiting. The role of the lysosomal and other proteases in Huntington is not clear. Some studies suggest that certain protease activities may contribute to toxicity whereas others are consistent with protection. These discrepancies may be due to a number of mechanisms including distinct effects of the specific intermediate digestion products of mutant huntingtin generated by different proteases. These observations suggested a critical need to investigate the consequence of upregulation of individual lysosomal enzyme in mutant huntingtin accumulation and toxicity. RESULTS: In this study, we used molecular approaches to enhance lysosomal protease activities and examined their effects on mutant huntingtin level and toxicity. We found that enhanced expression of lysosomal cathepsins D and B resulted in their increased enzymatic activities and reduced both full length and fragmented huntingtin in transfected HEK cells. Furthermore, enhanced expression of cathepsin D or B protected against mutant huntingtin toxicity in primary neurons, and their neuroprotection is dependent on macroautophagy. CONCLUSIONS: These observations demonstrate a neuroprotective effect of enhancing lysosomal cathepsins in reducing mutant huntingtin level and toxicity in transfected cells. They highlight the potential importance of neuroprotection mediated by cathepsin D or B through macroautophagy. PMID- 21631943 TI - Evaluation of medical student self-rated preparedness to care for limited English proficiency patients. AB - BACKGROUND: Patients with limited English proficiency (LEP) represent a growing proportion of the US population and are at risk of receiving suboptimal care due to difficulty communicating with healthcare providers who do not speak their language. Medical school curricula are required to prepare students to care for all patients, including those with LEP, but little is known about how well they achieve this goal. We used data from a survey of medical students' cross-cultural preparedness, skills, and training to specifically explore their self-rated preparedness to care for LEP patients. METHODS: We electronically surveyed students at one northeastern US medical school. We used bivariate analyses to identify factors associated with student self-rated preparedness to care for LEP patients including gender, training year, first language, race/ethnicity, percent LEP and minority patients seen, and skill with interpreters. We used multivariate logistic regression to examine the independent effect of each factor on LEP preparedness. In a secondary analysis, we explored the association between year in medical school and self-perceived skill level in working with an interpreter. RESULTS: Of 651 students, 416 completed questionnaires (63.9% response rate). Twenty percent of medical students reported being very well or well-prepared to care for LEP patients. Of these, 40% were in their fourth year of training. Skill level working with interpreters, prevalence of LEP patients seen, and training year were correlated (p < 0.001) with LEP preparedness. Using multivariate logistic regression, only student race/ethnicity and self-rated skill with interpreters remained statistically significant. Students in third and fourth years were more likely to feel skilled with interpreters (p < 0.001). CONCLUSIONS: Increasingly, medical students will need to be prepared to care for LEP patients. Our study supports two strategies to improve student preparedness: training students to work effectively with interpreters and increasing student diversity to better reflect the changing US demographics. PMID- 21631944 TI - MHC class I-related chain A and B ligands are differentially expressed in human cervical cancer cell lines. AB - BACKGROUND: Natural killer (NK) cells are an important resource of the innate immune system directly involved in the spontaneous recognition and lysis of virus infected and tumor cells. An exquisite balance of inhibitory and activating receptors tightly controls the NK cell activity. At present, one of the best characterized activating receptors is NKG2D, which promotes the NK-mediated lysis of target cells by binding to a family of cell surface ligands encoded by the MHC class I chain-related (MIC) genes, among others. The goal of this study was to describe the expression pattern of MICA and MICB at the molecular and cellular levels in human cervical cancer cell lines infected or not with human papillomavirus, as well as in a non-tumorigenic keratinocyte cell line. RESULTS: Here we show that MICA and MICB exhibit differential expression patterns among HPV-infected (SiHa and HeLa) and non-infected cell lines (C33-A, a tumor cell line, and HaCaT, an immortalized keratinocyte cell line). Cell surface expression of MICA was higher than cell surface expression of MICB in the HPV-positive cell lines; in contrast, HPV-negative cells expressed lower levels of MICA. Interestingly, the MICA levels observed in C33-A cells were overcome by significantly higher MICB expression. Also, all cell lines released higher amounts of soluble MICB than of soluble MICA into the cell culture supernatant, although this was most pronounced in C33-A cells. Additionally, Real-Time PCR analysis demonstrated that MICA was strongly upregulated after genotoxic stress. CONCLUSIONS: This study provides evidence that even when MICA and MICB share a high degree of homology at both genomic and protein levels, differential regulation of their expression and cell surface appearance might be occurring in cervical cancer-derived cells. PMID- 21631945 TI - Comparative genomics of four closely related Clostridium perfringens bacteriophages reveals variable evolution among core genes with therapeutic potential. AB - BACKGROUND: Because biotechnological uses of bacteriophage gene products as alternatives to conventional antibiotics will require a thorough understanding of their genomic context, we sequenced and analyzed the genomes of four closely related phages isolated from Clostridium perfringens, an important agricultural and human pathogen. RESULTS: Phage whole-genome tetra-nucleotide signatures and proteomic tree topologies correlated closely with host phylogeny. Comparisons of our phage genomes to 26 others revealed three shared COGs; of particular interest within this core genome was an endolysin (PF01520, an N-acetylmuramoyl-L-alanine amidase) and a holin (PF04531). Comparative analyses of the evolutionary history and genomic context of these common phage proteins revealed two important results: 1) strongly significant host-specific sequence variation within the endolysin, and 2) a protein domain architecture apparently unique to our phage genomes in which the endolysin is located upstream of its associated holin. Endolysin sequences from our phages were one of two very distinct genotypes distinguished by variability within the putative enzymatically-active domain. The shared or core genome was comprised of genes with multiple sequence types belonging to five pfam families, and genes belonging to 12 pfam families, including the holin genes, which were nearly identical. CONCLUSIONS: Significant genomic diversity exists even among closely-related bacteriophages. Holins and endolysins represent conserved functions across divergent phage genomes and, as we demonstrate here, endolysins can have significant variability and host specificity even among closely-related genomes. Endolysins in our phage genomes may be subject to different selective pressures than the rest of the genome. These findings may have important implications for potential biotechnological applications of phage gene products. PMID- 21631946 TI - Improving children's nutrition environments: a survey of adoption and implementation of nutrition guidelines in recreational facilities. AB - BACKGROUND: Although the mandate of recreational facilities is to enhance well being, many offer foods inconsistent with recommendations for healthy eating. Little is known regarding recreational facility food environments and how they might be improved, as few studies exist. The Alberta Nutrition Guidelines for Children and Youth (ANGCY) are intended to ensure access to healthy food choices in schools, childcare and recreational facilities. This study investigated awareness, adoption and implementation of the ANGCY among recreational facilities in Alberta, Canada, one year following their release. METHODS: A cross-sectional telephone survey was conducted from June - December, 2009 (n = 151) with managers of publicly funded recreational facilities that served food. The questionnaire included 10 closed and 7 open ended questions to assess the organizational priority for healthy eating, awareness, adoption and implementation of the ANGCY. Chi-squared tests examined quantitative variables, while qualitative data were analysed using directed content analysis. Greenhalgh's model of diffusion of complex innovations within health service organizations constituted the theoretical framework for the study. RESULTS: One half of respondents had heard of the ANGCY, however their knowledge of them was limited. Although 51% of facilities had made changes to improve the nutritional quality of foods offered in the past year, only a small fraction (11%) of these changes were motivated by the ANGCY. At the time of the survey, 14% of facilities had adopted the ANGCY and 6% had implemented them. Barriers to adoption and implementation were primarily related to perceived negative attributes of the ANGCY, the inner (organizational) context, and negative feedback received during the implementation process. Managers strongly perceived that implementing nutrition guidelines would limit their profit-making ability. CONCLUSIONS: If fully adopted and implemented, the ANGCY have the potential to make a significant and sustained contribution to improving the recreational facility food environment, however one year following their release, awareness, adoption and implementation of the ANGCY remained low. A mandated policy approach could offer an efficacious, cost-effective means of improving the food environment within recreational facilities. PMID- 21631947 TI - Effects of canola and corn oil mimetic on Jurkat cells. AB - BACKGROUND: The Western diet is high in omega-6 fatty acids and low in omega-3 fatty acids. Canola oil contains a healthier omega 3 to omega 6 ratio than corn oil. Jurkat T leukemia cells were treated with free fatty acids mixtures in ratios mimicking that found in commercially available canola oil (7% alpha linolenic, 30% linoleic, 54% oleic) or corn oil (59% linoleic, 24% oleic) to determine the cell survival or cell death and changes in expression levels of inflammatory cytokines and receptors following oil treatment. METHODS: Fatty acid uptake was assessed by gas chromatography. Cell survival and cell death were evaluated by cell cycle analyses, propidium-iodide staining, trypan blue exclusion and phosphatidylserine externalization. mRNA levels of inflammatory cytokines and receptors were assessed by RT-PCR. RESULTS: There was a significant difference in the lipid profiles of the cells after treatment. Differential action of the oils on inflammatory molecules, following treatment at non cytotoxic levels, indicated that canola oil mimetic was anti-inflammatory whereas corn oil mimetic was pro-inflammatory. SIGNIFICANCE: These results indicate that use of canola oil in the diet instead of corn oil might be beneficial for diseases promoted by inflammation. PMID- 21631948 TI - Dietary behaviors related to cancer prevention among pre-adolescents and adolescents: the gap between recommendations and reality. AB - BACKGROUND: Diet is thought to play an important role in cancer risk. This paper summarizes dietary recommendations for cancer prevention and compares these recommendations to the dietary behaviors of U.S. youth ages 8-18. METHODS: We identified cancer prevention-related dietary recommendations from key health organizations and assessed dietary consumption patterns among youth using published statistics from the National Health and Nutrition Examination Survey, the national Youth Risk Behavior Survey, and other supplemental sources. RESULTS: Cancer prevention guidelines recommend a diet rich in fruits, vegetables, and whole grains, recommend limiting sugary foods and beverages, red and processed meats, sodium, and alcohol, and recommend avoiding foods contaminated with carcinogens. However, youth typically do not meet the daily recommendations for fruit, vegetable, or whole grain consumption and are over-consuming energy-dense, sugary and salty foods. CONCLUSIONS: A large discrepancy exists between expert recommendations about diet and cancer and actual dietary practices among young people and points to the need for more research to better promote the translation of science into practice. Future research should focus on developing and evaluating policies and interventions at the community, state and national levels for aligning the diets of youth with the evolving scientific evidence regarding cancer prevention. PMID- 21631949 TI - Importance of pre-analytical steps for transcriptome and RT-qPCR analyses in the context of the phase II randomised multicentre trial REMAGUS02 of neoadjuvant chemotherapy in breast cancer patients. AB - BACKGROUND: Identification of predictive markers of response to treatment is a major objective in breast cancer. A major problem in clinical sampling is the variability of RNA templates, requiring accurate management of tumour material and subsequent analyses for future translation in clinical practice. Our aim was to establish the feasibility and reliability of high throughput RNA analysis in a prospective trial. METHODS: This study was conducted on RNA from initial biopsies, in a prospective trial of neoadjuvant chemotherapy in 327 patients with inoperable breast cancer. Four independent centres included patients and samples. Human U133 GeneChips plus 2.0 arrays for transcriptome analysis and quantitative RT-qPCR of 45 target genes and 6 reference genes were analysed on total RNA. RESULTS: Thirty seven samples were excluded because i) they contained less than 30% malignant cells, or ii) they provided RNA Integrity Number (RIN) of poor quality. Among the 290 remaining cases, taking into account strict quality control criteria initially defined to ensure good quality of sampling, 78% and 82% samples were eligible for transcriptome and RT-qPCR analyses, respectively. For RT-qPCR, efficiency was corrected by using standard curves for each gene and each plate. It was greater than 90% for all genes. Clustering analysis highlighted relevant breast cancer phenotypes for both techniques (ER+, PR+, HER2+, triple negative). Interestingly, clustering on trancriptome data also demonstrated a "centre effect", probably due to the sampling or extraction methods used in on of the centres. Conversely, the calibration of RT-qPCR analysis led to the centre effect withdrawing, allowing multicentre analysis of gene transcripts with high accuracy. CONCLUSIONS: Our data showed that strict quality criteria for RNA integrity assessment and well calibrated and standardized RT-qPCR allows multicentre analysis of genes transcripts with high accuracy in the clinical context. More stringent criteria are needed for transcriptome analysis for clinical applications. PMID- 21631950 TI - Cytoplasm affects grain weight and filled-grain ratio in indica rice. AB - BACKGROUND: Cytoplasmic effects on agronomic traits--involving cytoplasmic and nuclear genomes of either different species or different cultivars--are well documented in wheat but have seldom been demonstrated in rice (Oryza sativa L.). To detect cytoplasmic effects, we introgressed the nuclear genomes of three indica cultivars--Guichao 2, Jiangchengkugu, and Dianrui 449--into the cytoplasms of six indica cultivars - Dijiaowujian, Shenglixian, Zhuzhan, Nantehao, Aizizhan, and Peta. These 18 nuclear substitution lines were evaluated during the winter season of 2005 in Sanya, Hainan, China, and during the summer season of 2006 in Kunming, Yunnan, China. The effects of 6 cytoplasm sources, 3 nucleus sources, 2 locations and their interactions were estimated for plant height, panicle length, panicle number per plant, spikelet number per panicle, grain weight, filled-grain ratio, and yield per plot. RESULTS: For five of the seven traits, analysis of variance showed that there were no significant cytoplasmic effects or interactions involving cytoplasmic effects. The effect of cytoplasm on 1000-grain weight was highly significant. Mean 1000-grain weight over the two locations in four of the six cytoplasms clustered close to the overall mean, whereas plants with Nantehao cytoplasm had a high, and those with Peta cytoplasm a low mean grain weight. There was a highly significant three-way interaction affecting filled-grain ratio. At Sanya, cytoplasms varied in very narrow ranges within nuclear backgrounds. Strong cytoplasmic effects were observed only at Kunming and in only two of the three nuclear backgrounds; in the Jianchenkugu nuclear background, there was no evidence of strong cytoplasmic effects at either location. In the Dianrui 449 and Guichao 2 nuclear background evaluated at Kunming, filled-grain ratios of the six cytoplasms showed striking rank shifts CONCLUSIONS: We detected cytoplasmic variation for two agronomically important traits in indica rice. The cytoplasm source had a significant effect on grain weight across the two experimental locations. There was also a significant cytoplasmic effect on filled-grain ratio, but only in two of three nuclear background and at one of the two locations. The results extend our previous findings with japonica rice, suggesting that the selection of appropriate cytoplasmic germplasm is broadly important in rice breeding, and that cytoplasmic effects on some traits, such as filled-grain ratio, cannot be generalized; effects should be evaluated in the nuclear backgrounds of interest and at multiple locations. PMID- 21631951 TI - Trends in smokeless tobacco use in the us workforce: 1987-2005. AB - The primary aim was to examine whether increasing workplace smoking restrictions have led to an increase in smokeless tobacco use among US workers. Smokeless tobacco exposure increases the risk of oral cavity, esophageal, and pancreatic cancers, and stroke. The prevalence of smokeless tobacco use decreased from 1987 2000, except among men 25-44. While smokeless tobacco use has declined in the general population, it may be that the prevalence of smokeless tobacco use has increased among workers due to workplace smoking restrictions, which have been shown to have increased over the years. Using the most current nationally representative National Health Interview Survey (NHIS) data, we examined whether increasing workplace smoking restrictions have led to an increase in smokeless tobacco use among US workers (n = 125,838). There were no significant changes in smokeless tobacco use prevalence from 1987-2005 (pooled prevalence = 3.53%); rates also were lower in smoke free workplaces. Worker groups with high rates of smokeless tobacco use included farm workers (10.51%) and blue collar workers (7.26%). Results indicate that smokeless tobacco prevention strategies targeting particular worker groups are warranted. PMID- 21631952 TI - Immunohistochemical localization of hepatopancreatic phospholipase A2 in Hexaplex trunculus digestive cells. AB - BACKGROUND: Mammalian sPLA2-IB localization cell are well characterized. In contrast, much less is known about aquatic primitive ones. The aquatic world contains a wide variety of living species and, hence represents a great potential for discovering new lipolytic enzymes and the mode of digestion of lipid food. RESULTS: The marine snail digestive phospholipase A2 (mSDPLA2) has been previously purified from snail hepatopancreas. The specific polyclonal antibodies were prepared and used for immunohistochimical and immunofluorescence analysis in order to determine the cellular location of mSDPLA2. Our results showed essentially that mSDPLA2 was detected inside in specific vesicles tentatively named (mSDPLA2+) granules of the digestive cells. No immunolabelling was observed in secretory zymogene-like cells. This immunocytolocalization indicates that lipid digestion in the snail might occur in specific granules inside the digestive cells. CONCLUSION: The cellular location of mSDPLA2 suggests that intracellular phospholipids digestion, like other food components digestion of snail diet, occurs in these digestive cells. The hepatopancreas of H. trunculus has been pointed out as the main region for digestion, absorption and storage of lipids. PMID- 21631953 TI - Use of a total traffic count metric to investigate the impact of roadways on asthma severity: a case-control study. AB - BACKGROUND: This study had two principal objectives: (i) to investigate the relationship between asthma severity and proximity to major roadways in Perth, Western Australia; (ii) to demonstrate a more accurate method of exposure assessment for traffic pollutants using an innovative GIS-based measure that fully integrates all traffic densities around subject residences. METHODS: We conducted a spatial case-control study, in which 'cases' were defined as individuals aged under 19 years of age with more severe asthma (defined here as two or more emergency department contacts with asthma in a defined 5-year period) versus age- and gender-matched 'controls' with less severe asthma (defined here as one emergency department contact for asthma). Traffic exposures were measured using a GIS-based approach to determine the lengths of the roads falling within a buffer area, and then multiplying them by their respective traffic counts. RESULTS: We examined the spatial relationship between emergency department contacts for asthma at three different buffer sizes: 50 metres, 100 metres and 150 metres. No effect was noted for the 50 metre buffer (OR = 1.07; 95% CI: 0.91 1.26), but elevated odds ratios were observed with for crude (unadjusted) estimates OR = 1.21 (95% CI: 1.00-1.46) for 100 metre buffers and OR = 1.25 (95% CI: 1.02-1.54) for 150 metre buffers. For adjusted risk estimates, only the 150 metre buffer yielded a statistically significant finding (OR = 1.24; 95% CI:1.00 1.52). CONCLUSIONS: Our study revealed a significant 24% increase in the risk of experiencing multiple emergency department contacts for asthma for every log-unit of traffic exposure. This study provides support for the hypothesis that traffic related air pollution increases the frequency of health service contacts for asthma. This study used advanced GIS techniques to establish traffic-weighted buffer zones around the geocoded residential location of subjects to provide an accurate assessment of exposure to traffic emissions, thereby providing a quantification of the ranges over which pollutants may exert a health effect. PMID- 21631954 TI - P2X7 receptor activation ameliorates CA3 neuronal damage via a tumor necrosis factor-alpha-mediated pathway in the rat hippocampus following status epilepticus. AB - BACKGROUND: The release of tumor necrosis factor-alpha (TNF-alpha) appears depend on the P2X7 receptor, a purinergic receptor. In the present study, we addressed the question of whether P2X7 receptor-mediated TNF-alpha regulation is involved in pathogenesis and outcome of status epilepticus (SE). METHODS: SE was induced by pilocarpine in rats that were intracerebroventricularly infused with saline-, 2',3'-O-(4-benzoylbenzoyl)-adenosine 5'-triphosphate (BzATP), adenosine 5' triphosphate-2',3'-dialdehyde (OxATP), A-438079, or A-740003 prior to SE induction. Thereafter, we performed Fluoro-Jade B staining and immunohistochemical studies for TNF-alpha and NF-kappaB subunit phosphorylations. RESULTS: Following SE, P2X7 receptor agonist (BzATP) infusion increased TNF-alpha immunoreactivity in dentate granule cells as compared with that in saline-infused animals. In addition, TNF-alpha immunoreactivity was readily apparent in the mossy fibers, while TNF-alpha immunoreactivity in CA1-3 pyramidal cells was unaltered. However, P2X7 receptor antagonist (OxATP-, A-438079, and A-740003) infusion reduced SE-induced TNF-alpha expression in dentate granule cells. In the CA3 region, BzATP infusion attenuated SE-induced neuronal damage, accompanied by enhancement of p65-Ser276 and p65-Ser311 NF-kappaB subunit phosphorylations. In contrast, OxATP-, A-438079, and A-740003 infusions increased SE-induced neuronal death. Soluble TNF p55 receptor (sTNFp55R), and cotreatment with BzATP and sTNFp55R infusion also increased SE-induced neuronal damage in CA3 region. However, OxATP-, sTNFp55R or BzATP+sTNFp55R infusions could not exacerbate SE induced neuronal damages in the dentate gyrus and the CA1 region, as compared to BzATP infusion. CONCLUSIONS: These findings suggest that TNF-alpha induction by P2X7 receptor activation may ameliorate SE-induced CA3 neuronal damage via enhancing NF-kappaB p65-Ser276 and p65-Ser311 phosphorylations. PMID- 21631955 TI - Streamlined protein expression and purification using cleavable self-aggregating tags. AB - BACKGROUND: Recombinant protein expression and purification remains a fundamental issue for biotechnology. Recently we found that two short self-assembling amphipathic peptides 18A (EWLKAFYEKVLEKLKELF) and ELK16 (LELELKLKLELELKLK) can induce the formation of active protein aggregates in Escherichia coli (E. coli), in which the target proteins retain high enzymatic activities. Here we further explore this finding to develop a novel, facile, matrix-free protein expression and purification approach. RESULTS: In this paper, we describe a streamlined protein expression and purification approach by using cleavable self-aggregating tags comprising of one amphipathic peptide (18A or ELK16) and an intein molecule. In such a scheme, a target protein is first expressed as active protein aggregate, separated by simple centrifugation, and then released into solution by intein-mediated cleavage. Three target proteins including lipase A, amadoriase II and beta-xylosidase were used to demonstrate the feasibility of this approach. All the target proteins released after cleavage were highly active and pure (over 90% in the case of intein-ELK16 fusions). The yields were in the range of 1.6 10.4 MUg/mg wet cell pellet at small laboratory scale, which is comparable with the typical yields from the classical his-tag purification, the IMPACT-CN system (New England Biolabs, Beverly, MA), and the ELP tag purification scheme. CONCLUSIONS: This tested single step purification is capable of producing proteins with high quantity and purity. It can greatly reduce the cost and time, and thus provides application potentials for both industrial scale up and laboratorial usage. PMID- 21631956 TI - Coffee and tea consumption in relation to inflammation and basal glucose metabolism in a multi-ethnic Asian population: a cross-sectional study. AB - BACKGROUND: Higher coffee consumption has been associated with a lower risk of type 2 diabetes in cohort studies, but the physiological pathways through which coffee affects glucose metabolism are not fully understood. The aim of this study was to evaluate the associations between habitual coffee and tea consumption and glucose metabolism in a multi-ethnic Asian population and possible mediation by inflammation. METHODS: We cross-sectionally examined the association between coffee, green tea, black tea and Oolong tea consumption and glycemic (fasting plasma glucose, HOMA-IR, HOMA-beta, plasma HbA1c) and inflammatory (plasma adiponectin and C-reactive protein) markers in a multi-ethnic Asian population (N = 4139). RESULTS: After adjusting for multiple confounders, we observed inverse associations between coffee and HOMA-IR (percent difference: - 8.8% for >= 3 cups/day versus rarely or never; Ptrend = 0.007), but no significant associations between coffee and inflammatory markers. Tea consumption was not associated with glycemic markers, but green tea was inversely associated with plasma C-reactive protein concentrations (percent difference: - 12.2% for >= 1 cup/day versus < 1 cup/week; Ptrend = 0.042). CONCLUSIONS: These data provide additional evidence for a beneficial effect of habitual caffeinated coffee consumption on insulin sensitivity, and suggest that this effect is unlikely to be mediated by anti inflammatory mechanisms. PMID- 21631957 TI - Two-step intensity modulated arc therapy (2-step IMAT) with segment weight and width optimization. AB - BACKGROUND: 2-step intensity modulated arc therapy (IMAT) is a simplified IMAT technique which delivers the treatment over typically two continuous gantry rotations. The aim of this work was to implement the technique into a computerized treatment planning system and to develop an approach to optimize the segment weights and widths. METHODS: 2-step IMAT was implemented into the Prism treatment planning system. A graphical user interface was developed to generate the plan segments automatically based on the anatomy in the beam's-eye-view. The segment weights and widths of 2-step IMAT plans were subsequently determined in Matlab using a dose-volume based optimization process. The implementation was tested on a geometric phantom with a horseshoe shaped target volume and then applied to a clinical paraspinal tumour case. RESULTS: The phantom study verified the correctness of the implementation and showed a considerable improvement over a non-modulated arc. Further improvements in the target dose uniformity after the optimization of 2-step IMAT plans were observed for both the phantom and clinical cases. For the clinical case, optimizing the segment weights and widths reduced the maximum dose from 114% of the prescribed dose to 107% and increased the minimum dose from 87% to 97%. This resulted in an improvement in the homogeneity index of the target dose for the clinical case from 1.31 to 1.11. Additionally, the high dose volume V105 was reduced from 57% to 7% while the maximum dose in the organ-at-risk was decreased by 2%. CONCLUSIONS: The intuitive and automatic planning process implemented in this study increases the prospect of the practical use of 2-step IMAT. This work has shown that 2-step IMAT is a viable technique able to achieve highly conformal plans for concave target volumes with the optimization of the segment weights and widths. Future work will include planning comparisons of the 2-step IMAT implementation with fixed gantry intensity modulated radiotherapy (IMRT) and commercial IMAT implementations. PMID- 21631958 TI - How important is the land use mix measure in understanding walking behaviour? Results from the RESIDE study. AB - BACKGROUND: Understanding the relationship between urban design and physical activity is a high priority. Different representations of land use diversity may impact the association between neighbourhood design and specific walking behaviours. This study examined different entropy based computations of land use mix (LUM) used in the development of walkability indices (WIs) and their association with walking behaviour. METHODS: Participants in the RESIDential Environments project (RESIDE) self-reported mins/week of recreational, transport and total walking using the Neighbourhood Physical Activity Questionnaire (n = 1798). Land use categories were incrementally added to test five different LUM models to identify the strongest associations with recreational, transport and total walking. Logistic regression was used to analyse associations between WIs and walking behaviour using three cut points: any (> 0 mins), >= 60 mins and >= 150 mins walking/week. RESULTS: Participants in high (vs. low) walkable neighbourhoods reported up to almost twice the amount of walking, irrespective of the LUM measure used. However, different computations of LUM were found to be relevant for different types and amounts of walking (i.e., > 0, >= 60 or >= 150 mins/week). Transport walking (>= 60 mins/week) had the strongest and most significant association (OR = 2.24; 95% CI:1.58-3.18) with the WI when the LUM included 'residential', 'retail', 'office', 'health, welfare and community', and 'entertainment, culture and recreation'. However, any (> 0 mins/week) recreational walking was more strongly associated with the WI (OR = 1.36; 95% CI:1.04-1.78) when land use categories included 'public open space', 'sporting infrastructure' and 'primary and rural' land uses. The observed associations were generally stronger for >= 60 mins/week compared with > 0 mins/week of transport walking and total walking but this relationship was not seen for recreational walking. CONCLUSIONS: Varying the combination of land uses in the LUM calculation of WIs affects the strength of relationships with different types (and amounts) of walking. Future research should examine the relationship between walkability and specific types and different amounts of walking. Our results provide an important first step towards developing a context-specific WI that is associated with recreational walking. Inherent problems with administrative data and the use of entropy formulas for the calculation of LUM highlight the need to explore alternative or complimentary measures of the environment. PMID- 21631960 TI - The importance of senescence in ionizing radiation-induced tumour suppression. AB - Cellular senescence is a condition of longlasting proliferation arrest, induced in cells in response to various stressors. These stressors include telomere shortening and/or dysfunction, DNA damage, and oncogene signalling. Epithelial and mesenchymal cells and also tumour cells derived from these tissues are more resistant to radiation-induced apoptosis and respond to irradiation mainly by senescence. Senescence-associated molecular mechanisms related to the activation of canonical DNA damage pathway ATM-p53 as well as mechanisms related to the extracellular signals, cytokine increase and upregulation of their receptors are discussed in this review. PMID- 21631959 TI - Comparative thermodynamic studies on substrate and product binding of O acetylserine sulfhydrylase reveals two different ligand recognition modes. AB - BACKGROUND: The importance of understanding the detailed mechanism of cysteine biosynthesis in bacteria is underscored by the fact that cysteine is the only sulfur donor for all cellular components containing reduced sulfur. O acetylserine sulfhydrylase (OASS) catalyzes this crucial last step in the cysteine biosynthesis and has been recognized as an important gene for the survival and virulence of pathogenic bacteria. Structural and kinetic studies have contributed to the understanding of mechanistic aspects of OASS, but details of ligand recognition features of OASS are not available. In the absence of any detailed study on the energetics of ligand binding, we have studied the thermodynamics of OASS from Salmonella typhimurium (StOASS), Haemophilus influenzae (HiOASS), and Mycobacterium tuberculosis (MtOASS) binding to their substrate O-acetylserine (OAS), substrate analogue (methionine), and product (cysteine). RESULTS: Ligand binding properties of three OASS enzymes are studied under defined solution conditions. Both substrate and product binding is an exothermic reaction, but their thermodynamic signatures are very different. Cysteine binding to OASS shows that both enthalpy and entropy contribute significantly to the binding free energy at all temperatures (10-30 degrees C) examined. The analyses of interaction between OASS with OAS (substrate) or methionine (substrate analogue) revealed a completely different mode of binding. Binding of both OAS and methionine to OASS is dominated by a favorable entropy change, with minor contribution from enthalpy change (DeltaH(St-Met) = -1.5 +/- 0.1 kJ/mol; TDeltaS(St-Met) = 8.2 kJ/mol) at 20 degrees C. Our salt dependent ligand binding studies indicate that methionine binding affinity is more sensitive to [NaCl] as compared to cysteine affinity. CONCLUSIONS: We show that OASS from three different pathogenic bacteria bind substrate and product through two different mechanisms. Results indicate that predominantly entropy driven methionine binding is not mediated through classical hydrophobic binding, instead, may involve desolvation of the polar active site. We speculate that OASS in general, may exhibit two different binding mechanisms for recognizing substrates and products. PMID- 21631961 TI - In vitro models for adipose tissue engineering with adipose-derived stem cells using different scaffolds of natural origin. AB - Soft tissue regeneration with cell and tissue engineering-based approaches has numerous potential applications in plastic and reconstructive surgery. Adipose derived stem cells (ASC) have been proved as a feasible source for adipose tissue engineering as they possess high proliferative and differentiation capacity. The purpose of our study was to evaluate adipogenic differentiation of human ASC in four different 3D scaffolds of natural origin, namely human platelet-poor plasma, alginate, fibrin gel and collagen sponge, to define their suitability for adipose tissue engineering and potential clinical applications. ASC were isolated from lipoaspirates of three adult female patients, seeded in the scaffolds, and adipogenic differentiation was induced. After two weeks of cultivation, the constructs were assessed for their mechanical and handling properties, cell viability and adipogenic differentiation. Additionally, the expression of vascular endothelial growth factor (VEGF) was analysed in different culture systems. The results indicate that the levels of specific adipogenic markers and VEGF expression were increased in 3D cultures, as compared to 2D culture. Among 3D scaffolds, fibrin gel showed optimal combination of mechanical characteristics and support of adipogenic differentiation; it was easy to handle, allowed high cell viability, and at the same time supported adipogenic differentiation and VEGF expression. PMID- 21631962 TI - In vitro effects of imatinib on glucose-6-phosphate dehydrogenase and glutathione reductase. AB - Imatinib (Gleevec, STI571) is a drug used to treat certain types of cancer. Glucose-6-phosphate dehydrogenase and glutathione reductase are enzymes important for redox homeostasis and play key roles in many cellular processes. The purpose of the present work is to evaluate the in vitro effects of imatinib on sheep brain cortex glucose-6-phosphate dehydrogenase, and on bovine kidney cortex, bovine liver and yeast glutathione reductase. Kinetic studies on the inhibition of enzymes by imatinib have been investigated by using Lineweaver-Burk double reciprocal plot and values summarized with graphs by plotting the data using Linewear-Burk diagrams of 1/v against 1/[S] at each [I]. Imatinib inhibits glucose- 6-phosphate dehydrogenase with an IC50 value of 0.7 mM. It inhibits bovine kidney cortex, liver and yeast glutathione reductase in a concentration dependent manner with IC50 values of 0.8, 0.92, 1 mM, respectively. We have investigated the kinetic characteristics, inhibition types and constants (Ki). Inhibition of the glucose-6-phosphate dehydrogenase and glutathione reductase represents an attractive approach to the development of anticancer agents. This study shows the molecular effectiveness of the drug on purified enzymes of various sources. Understanding the kinetic mechanism of the drug and enzyme relationship may be a powerful approach to the future drug studies concerning new cancer drugs, drug resistance and new aspects in cancer therapy. PMID- 21631963 TI - DNA analysis of renal electrolyte transporter genes among patients suffering from Bartter and Gitelman syndromes: summary of mutation screening. AB - Patients with renal diseases associated with salt-losing tubulopathies categorized as Gitelman and classic form of Bartter syndrome have undergone genetic screening for possible mutation capture in two different genes: SLC12A3 and CLCNKB. Clinical symptoms of these two diseases may overlap. Patients with clinical symptoms of antenatal form of Bartter syndrome were screened for mutations in two different genes: KCNJ1 and SLC12A1. The aim was to establish genetic mutation screening of Bartter/Gitelman syndrome and to confirm the proposed diagnosis. We have identified seven different causative mutations in the SLC12A3 gene, four in the CLCNKB gene, two in the SLC12A1 gene, and none in the KCNJ1 gene. Nine of these mutations are novel. In one case, genetic analysis led to re-evaluation of diagnosis between the Gitelman and classic form of Bartter syndrome. PMID- 21631964 TI - The effect of ATM and ERK1/2 inhibition on mitoxantrone-induced cell death of leukaemic cells. AB - The relationship between signal pathways MEK1/2-ERK1/2 and ATM-p53 in the response to DNA damage is not well understood. The aim of our study was to investigate the effect of mitoxantrone and two protein kinase inhibitors - caffeine (inhibitor of ATM kinase) and U0126 (inhibitor of MEK1/2 kinase) - on MOLT-4 and Jurkat leukaemic cell lines. In this work we show that the inhibition of MEK1/2 is associated with an increased mortality of cells after mitoxantrone treatment. Inhibition of ATM by caffeine delayed mitoxantrone-induced cell death in MOLT-4 cells. Mitoxantrone itself induced cell-cycle arrest and accumulation of the cells in late S and G2/M phase. Inhibition of ATM, but not of MEK1/2, abrogated mitoxantrone-induced cell-cycle arrest. Inhibition of MEK1/2 did not change mitoxantroneinduced up-regulation of p53 and p21, but inhibition of ATM markedly decreased up-regulation of p53 and p21, and p53 phosphorylation on serine 15 and serine 392. It can be concluded that: 1) mitoxantrone- induced phosphorylation of p53 on serine 15 and serine 392 is ATM dependent and MEK1/2 ERK1/2 independent. 2) ATM inhibition by caffeine prevents G2 cell arrest and in p53-positive cells MOLT-4 delays the onset of mitoxantrone-induced cell death. 3) Inhibition of MEK1/2-ERK1/2 cascade potentiates the cytostatic effect of mitoxantrone regardless of the p53 status. PMID- 21631965 TI - Pregnancy-associated plasma protein A polymorphisms in patients with risk pregnancies. AB - Pregnant women are often threatened by hypertension, symptoms of preterm labour, hepatopathy, and other. These complications might be the consequence of genetic factors together with involvement of environmental factors. We were searching for three polymorphisms Arg654Lys, Ala678Pro and Thr686Ala in exon 5, and two polymorphisms Phe802Leu, Ser827Ser/Leu in exon 7, and for the new mutations in exons 5 and 7 of the pregnancy-associated plasma protein A gene in the studied group consisting of 203 women - 79 pregnant women in time of preterm labour, 24 pregnant women suffering from preeclampsia, and 100 healthy pregnant and non pregnant women serving as controls. We did not find any divergence from wild-type form of these polymorphisms in any of the studied groups, which led us to the hypothesis that these polymorphisms are not associated with our studied group of Caucasian origin. However, further studies with a larger group of subjects are needed to confirm our results. PMID- 21631966 TI - Sex-specific associations of nutrition with hypertension and systolic blood pressure in Alaska Natives findings from the GOCADAN study. AB - OBJECTIVES: To examine sex-specific associations of nutritional factors with prevalent hypertension (HTN) and systolic blood pressure (SBP) in Alaska Natives. Diet is known to affect SBP, a major risk factor for cardiovascular disease. STUDY DESIGN: Cross-sectional analysis of participants without diabetes in the Genetics of Coronary Artery Disease in Alaska Natives study. METHODS: Macronutrients such as fat, carbohydrate and protein and micronutrients such as sodium were investigated. HTN was defined as SBP>=140 mmHg, diastolic blood pressure>=90 mmHg and/or taking anti-HTN medication. Analyses were stratified by sex and covariates included age, body mass index (BMI), energy intake, smoking and physical activity. RESULTS: Mean age was 42 years for men (n=456) and women (n=602). Men with HTN (n=106) compared to men without HTN consumed a higher proportion of calories from total (p=0.01), saturated (p<0.01) and trans fatty acid (p=0.03) fats. Women with HTN (n=99) compared to women without HTN consumed more total (p=0.03) and monounsaturated (p=0.04) fat, higher protein (p=0.02) and lower total (p<0.01) and simple (p<0.01) carbohydrates. After covariate adjustment, men not on anti-HTN medications (n=407) had significantly higher average SBP with increasing quartiles of trans fatty acid intake (p for linear trend=0.01) and sodium intake (p for linear trend=0.02). For women not on anti HTN medications (n=528), after covariate adjustment, average SBP decreased with increasing quartiles of omega 3 fatty acid intake (p for linear trend <0.01). CONCLUSIONS: Prospective evaluation of the sex-specific associations of nutritional factors with HTN and SBP on outcomes is needed along with novel interventions to lower the risk of cardiovascular disease. PMID- 21631967 TI - Importance of traditional foods for the food security of two First Nations communities in the Yukon, Canada. AB - OBJECTIVES: This study sought to evaluate food consumption patterns in the context of food security in two Yukon First Nations communities. STUDY DESIGN: Twenty-nine members of Vuntut Gwitchin households in Old Crow and 33 members of Tlingit households in Teslin participated in individual interviews. METHODS: Food frequency questionnaires were used to quantify traditional food consumption throughout the spring 2007 and winter 2008 and to identify potential temporal trends through a comparison with data from the early 1990s. Additional questions, including the Health Canada Household Food Security Survey Module, sought to assess food security concerns in each community. RESULTS: Overall frequency of traditional food consumption did not change in either community from the 2 time point analyses. There was, however, a difference in frequency of consumption of certain groups of foods, and this highlighted the degree to which environmental variability affects the availability of foods. CONCLUSION: The importance of traditional foods in the diet of Yukon First Nations has not changed over the past 15 years. However, limited availability of food species, access to harvesting equipment and decrease in available time to go out on the land to harvest are food security challenges facing households today. PMID- 21631968 TI - Diet and lifestyle of the Sami of southern Lapland in the 1930s--1950s and today. AB - OBJECTIVES: To describe the lifestyle of the Sami of southern Lapland 50 to 70 years ago in relation to the present-day Sami and non-Sami populations and, thereby, to provide a basis for future studies of culturally related determinants of health and illness. STUDY DESIGN: A qualitative analysis, and a quantitative comparison of Sami and non-Sami groups. METHODS: Semi-structured interviews were conducted with 20 elderly Sami concerning their parents' lifestyle and diet 50 to 70 years ago. Questionnaire data from 81 reindeer-herding Sami, 226 non-reindeer herding Sami and 1,842 sex-, age- and geographically matched non-Sami from the population-based Vasterbotten Intervention Project were analysed by non parametric tests and partial least squares methodology. RESULTS: Surprisingly, fatty fish may have been more important than reindeer meat for the Sami of southern Lapland in the 1930s to 1950s, and it is still consumed more frequently by reindeer-herding Sami than nonreindeer-herding Sami and non-Sami. Other dietary characteristics of the historical Sami and present-day reindeer-herding Sami were higher intakes of fat, blood and boiled coffee, and lower intakes of bread, fibre and cultivated vegetables, compared with present-day non-Sami. Physical activity was also a part of the daily life of the Sami to a greater extent in the 1930s to 1950s than today. Sami men often worked far from home, while the women were responsible for fishing, farming, gardening (which was introduced in the 1930-1950 period), as well as housework and childcare. CONCLUSIONS: For studies investigating characteristic lifestyle elements of specific ethnic groups, the elements of greatest acknowledged cultural importance today (in this case reindeer meat) may not be of the most objective importance traditionally. PMID- 21631969 TI - Hospital General de Mexico: 105 years since its foundation. PMID- 21631970 TI - Evaluation of bilateral cingulotomy and anterior capsulotomy for the treatment of aggressive behavior. AB - BACKGROUND: Agressiveness is a psychiatric symptom that may be part of schizophrenia, mental retardation, drug abuse and other conditions. Surgical treatment remains controversial and few therapeutic options are available. We undertook this study to perform a prospective analysis on the efficacy and safety of bilateral cingulotomy and anterior capsulotomy in the treatment of aggressiveness behavior. METHODS: We studied 25 patients with a primary diagnosis of aggressiveness refractory to conventional treatment. Subjects were clinically evaluated with the Mayo-Portland adaptability inventory and the Global Assessment of Functioning score. Lesions were placed stereotactically in both targets and confirmed by postoperative magnetic resonance imaging. Significant changes were evaluated with Wilcoxon test after 3 and 6 months. RESULTS: According to inclusion and exclusion criteria, only 12 patients were finally included and surgical treated. Lesions significantly decreased using the Mayo-Portland adaptability inventory and the Global Assessment of Functioning score (p <0.002) at 3 and 6 months follow-up. Only five patients showed either mild or transitory postsurgical complications. CONCLUSIONS: Combined bilateral anterior capsulotomy and cingulotomy successfully reduced aggressiveness behavior and improved clinical evaluations. These effects were obtained with fewer complications than previously described targets. PMID- 21631971 TI - Prevalence of thyroid nodules in the Valley of Mexico. AB - BACKGROUND: There are no open population epidemiological studies to determine the true prevalence of thyroid nodules in the Valley of Mexico. The aim of this investigation was to determine prevalence. METHODS: We carried out a cross sectional, observational, descriptive study in 2401 open population subjects in the Valley of Mexico. All subjects were at least 18 years of age with no clinical history of thyroid disease. Variables analyzed were age, gender, thyroid nodule identified by neck palpation or neck ultrasound, TSH levels, cytological diagnosis and histopathological diagnosis in case of clinical thyroid nodule detection. Percentages as a summary measure for qualitative variables and proportions and chi-square for independent variables were used for statistical analysis. RESULTS: Included were 2401 subjects with a mean age of 39 years (82.4% female, 17.6% male). Thirty-four (1.4%) thyroid nodules were detected by palpation and 471 (19.6%) by ultrasound. The 34 palpable thyroid nodules corresponded to 16 (47.2%) colloid goiters, 8 (23.5%) follicular adenomas, 7 (20.5%) Hashimoto's thyroiditis, 2 (5.9%) papillary carcinomas and 1 (2.9%) oxiphilic cell adenoma. Nonpalpable thyroid nodules detected by ultrasound did not develop illness at 2-year follow-up. TSH was normal in 1620 (67.4%), high in 515 (21.5%) and low in 266 (11.1%) patients. There was no relationship between TSH levels and final diagnosis of the 34 palpable nodules (p >0.05). CONCLUSIONS: Prevalence of palpable thyroid nodule in the Valley of Mexico is 1.4%; 5.9% of these are due to papillary thyroid cancer. TSH levels do not correlate with the presence or cause of the thyroid nodule. PMID- 21631972 TI - Low-risk papillary thyroid cancer recurrence in patients treated with total thyroidectomy and adjuvant therapy vs. patients treated with partial thyroidectomy. AB - BACKGROUND: Surgical extension for treatment of patients with low-risk papillary thyroid carcinoma is still controversial. We undertook this study to assess if there is a difference in recurrence between patients undergoing total thyroidectomy plus adjuvant therapy and patients treated with only partial thyroidectomy. METHODS: We conducted a longitudinal, observational, analytical study in patients with histopathological diagnosis of low-risk papillary thyroid carcinoma followed for at least 10 years. Patients were divided into two groups: Group 1: Patients treated with total thyroidectomy plus adjuvant therapy (TT) and Group 2: Patients treated with only partial thyroidectomy without adjuvant therapy (HT). Descriptive and inferential statistical methods were used. RESULTS: AMES: 184 patients, recurrence in 5/23 HT and 7/161 TT (p = 0.0016); MACIS: 170 patients, recurrence in 5/24 HT and 5/146 TT (p = 0.0008); DeGroot: 92 patients, recurrence in 3/19 HT and 2/73 TT (p = 0.0254); TNM: 150 patients, recurrence in 5/22 HT and 7/128 TT (p = 0.0058). The time interval for local recurrences was higher in comparison to regional recurrences (p <0.05). In all classifications, recurrences occur mainly with regional metastatic disease (60%). Multifocality, bilateral disease and extracapsular disease showed no statistical difference. There was one incidental injury to a recurrent laryngeal nerve and this was repaired during the same surgical procedure. There was no morbidity due to hypoparathyroidism. CONCLUSIONS: Patients classified as low risk according to any of the studied classifications have a higher risk for recurrence when treated with hemithyroidectomy than when treated with total thyroidectomy plus adjuvant therapy. PMID- 21631973 TI - Elevated triglycerides/HDL-cholesterol ratio associated with insulin resistance. AB - BACKGROUND: Cardiovascular disease is the main cause of death worldwide and insulin resistance (IR) plays an important role for its development. In addition, IR has been associated with hypertension and metabolic syndrome according to the triglyceride/HDL-cholesterol (TGL/HDL) ratio. We undertook this study to determine whether the TGL/HDL ratio is associated with IR in apparently healthy subjects. METHODS: A cross-sectional study including healthy men and nonpregnant women was performed. Individuals with IR were compared against subjects without IR. Variables studied were age, gender, body mass index, and waist circumference. Exclusion criteria were chronic diseases such as renal disease, hepatic disease, malignancy, and diabetes. RESULTS: A total of 177 subjects were enrolled, 117 females (66.1%) and 60 males (33.9%). Of these, 145 (93 females and 52 males) with IR were compared against 32 subjects (24 females and 8 males) without IR. Elevated ratio TGL/HDL ratio was detected in 89 (61.4%) and 12 (38.6%) subjects with and without IR, respectively. The elevated TGL/HDL ratio was significantly associated with IR (OR 2.64, 95% CI = 1.12-6.29). CONCLUSIONS: In apparently healthy subjects, elevated TGL/HDL ratio was significantly associated with the presence of IR. PMID- 21631974 TI - Biochemical analysis and lipid peroxidation in liver ischemic preconditioning. AB - BACKGROUND: Hepatic surgery requires, under diverse circumstances, periods of ischemia and reperfusion (I-R) such as those present in liver resection, hepatic injury, and liver transplantation. The objective of the present work was to conduct an experimental study to evaluate the effect of hepatic preconditioning (HPC) on modulation of the I-R injury. METHODS: Male Wistar rats were distributed into the following three study groups: group 1, simulated or sham; group 2, submitted to a 30-min period of total warm ischemia and a reperfusion phase, and group 3, in which we carried out 10-min preconditioning of warm ischemia and 10 min of reperfusion prior to the total ischemia period for a total of 60 min and the reperfusion phase. We obtained liver biopsies for thiobarbituric acid reactive substances (TBARS) (MDA, thiobarbituric acid adducts) and for blood sample determinations in serum of liver-cell enzymes such as alanine aminotransferase and aspartate aminotransferase during a 24-h time course. RESULTS: We observed a decrease in the variables studied in group 3 (HPC) as well as of serum liver enzymes and TBARS levels such as indirect oxidative stress indicators upon comparison with group 2 animals submitted to total hepatic I-R. CONCLUSIONS: HPC is an efficient surgical strategy for decreasing the elevation of hepatic enzymes and indirect lipoperoxidation indicators in an I-R model. Controlled clinical studies should be performed to determine its functional properties and clinical applicability. PMID- 21631975 TI - Factors associated with postoperative complications and mortality in perforated peptic ulcer. AB - BACKGROUND: Elective surgery for uncomplicated peptic ulcer has shown a significant decrease; however, complications such as perforation and obstruction persist and require urgent surgical management. The aim of this study was to identify factors associated with early postoperative complications and mortality of patients admitted to the emergency department with perforated peptic ulcer. METHODS: We performed a clinical, retrospective, cross-sectional and descriptive study of patients who were treated at the General Hospital of Mexico with a diagnosis of perforated peptic ulcer from January 2006 to December 2008. Thirty patients were included in the study. We studied several clinical findings upon admission to the emergency department and intraoperative patient findings in order to determine the association of those with early postoperative complications and mortality. RESULTS: We studied 30 patients with an average age of 57.07 years (+/- 14.2 years). The male:female ratio was 2:1. We found that the risk of developing postoperative complications was 66.7% and is significantly influenced by time of onset of abdominal pain prior to admission, bloating, septic shock and blood type O positive. Mortality was 16.7% and was correlated with the presence of septic shock on admission. The surgical procedure performed was primary closure with Graham patch in 86.6%. Average hospital stay was 12.8 days. CONCLUSIONS: The presence of early postoperative complications is associated with time of onset of abdominal pain before admission, abdominal distension, blood type O positive and the presence of septic shock on admission. PMID- 21631976 TI - Quality of life in ostomized patients. AB - BACKGROUND: Ostomized patients usually have some concerns such as absence of sphincter control, noisy bowel movements, changes in passage of gas, social discomfort due to odors, concerns about bag filling, peristomal and skin irritation as well as psychological changes such as body image distortion, among others, that will determine quality of life. We undertook this study to measure quality of life in ostomized patients attending the Coloproctology Unit in Hospital General de Mexico. METHODS: We carried out a cross-sectional descriptive study from January 1, 2009 to May 29, 2009 in ostomized patients. Two scales were used to determine quality of life. The first was quality of life SF-36 (version 2) with 36 items that measured eight aspects. The other scale used was the Montreux scale. RESULTS: For the Montreux scale, quality of life section, a range between 60.8 and 81.2 points was obtained. The results obtained with the quality of life scale SF36 were as follows: physical function, 67.04; physical role, 56.06; physical pain, 68.28; general health, 57.16; with an overall average of 62.14 points. For mental aspects, results were as follows: vitality, 68.37; social function, 71.53; emotional role, 71.48; and mental health, 68.67; with an overall average of 70.01 points. CONCLUSIONS: Quality of life of subjects included was classified as good. Fear of stoma leaks and sexual function were the aspects that most affected the patients, and those are responsible for a decrease in quality of life in this study. PMID- 21631977 TI - Assessment of medical residents' satisfaction. AB - BACKGROUND: Modern medical education is focused on students, and it is necessary to assess its level of satisfaction. A questionnaire was validated and we then conducted a study about the educational satisfaction level of medical residents of the Hospital General of Mexico. METHODS: An observational, descriptive, cross sectional and prospective study was conducted. A questionnaire of 21 items was validated and then applied to a representative sample of medical residents. Each item was evaluated with a scale from 0 to 10 and then gathered in groups: 0-5 = poor, 6-7 = average, 8 = good, 9 = very good, and 10 = excellent. Descriptive and inferential statistics were carried out using SPSS v.17.0. RESULTS: The questionnaire had internal validity with Cronbach's alpha >0.91 by item. Included in the study were 355 medical residents representing 37 different specialties. The performance perception of the iheadi professors showed a wide heterogeneity: excellent (23.7%), very good (20.6%), good (16.9%), average (23.1%), poor (15.8%). Fourth-year residents and upward valued the educational performance higher (p = 0.001) as well as medical/surgical residents (p = 0.02). Intermediate level residents valued the professor higher (p = 0.001), similar to students who were married or living with a partner (p <0.001). Upon contrasting the evaluation of the teacher's performance with the overall course performance, a linear, direct and significant correlation was obtained with Spearman's correlation coefficient = 0.78 and regression coefficient (p <0.001). CONCLUSIONS: We found a wide range of heterogeneity of results. Performance of the professors was the basic component to judge the quality of the residents' courses. PMID- 21631978 TI - Global cardiovascular risk stratification: comparison of the Framingham method with the SCORE method in the Mexican population. AB - BACKGROUND: In the Mexican population we are unaware if the Framingham model is a better system than the SCORE system for stratifying cardiovascular risk. The present study was conducted to compare risk stratification with the Framingham tables using the same procedure but using the SCORE, with the aim of recommending the use of the most appropriate method. METHODS: We analyzed a database of apparently healthy workers from the Mexico City General Hospital included in the study group "PRIT" (Prevalencia de Factores de Riesgo de Infarto del Miocardio en Trabajadores del Hospital General de Mexico) and we calculated the risk in each simultaneously with the Framingham method and the SCORE method. RESULTS: It was possible to perform risk calculation with both methods in 1990 subjects from a total of 5803 PRITHGM study participants. When using the SCORE method, we stratified 1853 patients into low risk, 133 into medium risk and 4 into high risk. The Framingham method qualified 1586 subjects as low risk, 268 as medium risk and 130 as high risk. Concordance between scales to classify both patients according to the same risk was 98% in those classified as low risk, 19.4% among those classified as intermediate risk and only 3% in those classified as high risk. CONCLUSIONS: According to our results, it seems more appropriate in our country to recommend the Framingham model for calculating cardiovascular risk due to the fact that the SCORE model underestimated risk. PMID- 21631979 TI - Impact of waist circumference reduction on cardiovascular risk in treated obese subjects. AB - BACKGROUND: The World Health Organization reports that waist circumference (WC) independent of weight or body mass index (BMI) predicts cardiovascular risk. We undertook this study to determine the change of prevalence in comorbidities associated with obesity and cardiovascular risk after favorably modifying WC. METHODS: We studied 153 nondiabetic patients with obesity (BMI =30 kg/m2) and WC in women =80 cm and in men =94 cm who entered a weight control program for 2 years. We evaluated the evolution of their anthropometric measurements and metabolic status. Ninety patients (58.8%) completed the study. With the prior acceptance of the patients, they received nutritional advice and psychological and physical activity support during their monthly visits. Also, anthropometric measurements and blood pressure were evaluated. At the beginning and after each 6 months, glucose, total cholesterol, HDL cholesterol and triglycerides were determined. At the beginning and at the end of study the Framingham risks were evaluated. RESULTS: Of the 90 patients, 37 (group 1) decreased their WC: in women <80 cm and in men <94 cm. In 53 patients (group 2) there were no significant changes. Changes were shown in group 1 for blood pressure (from 36.6% to 21.6%), hyperglycemia >100 mg/dl decreased from 18.8% to 8.1%, triglycerides >150 mg/dl decreased from 28.8% to 18.9% and Framingham risk at 10 years decreased. CONCLUSIONS: There is a direct relationship between WC and cardiovascular risk. When WC decreases, cardiovascular risk is favorably modified. Measurement of WC is a good predictor of cardiovascular risk. PMID- 21631980 TI - Complex localization of single-rod contraceptive implant. Report of two clinical cases. AB - BACKGROUND: The subdermal single-rod contraceptive implant is used by more than one million women worldwide. In Mexico there are ~ 600,000 colocated implants. Cases of complex implant localization caused by a deep insertion procedure have been reported. CLINICAL CASES: Two clinical cases of implant with complex localization are presented. Case 1: we present a 21-year-old female. After the insertion procedure during the 12-, 24-, and 36-month revisions, the implant could not be located. Case 2: we present a 28-year-old female with subdermal single-rod contraceptive implant co-located in the external side of the left arm, partially palpable on the extreme distal area. Transverse cut of ultrasound showed the extreme distal area of the implant at 6 and 7 cm, respectively, at the site of insertion (scar). CONCLUSIONS: Due to simplicity and accessibility, ultrasound is the selected method for identifying deep nonpalpable implants. PMID- 21631981 TI - Duodeno-biliary obstruction in Peutz-Jeghers syndrome. AB - BACKGROUND: Peutz-Jeghers syndrome (PJS) is a pathology with autosomal dominant inheritance characterized by the presence of hamartomatous polyposis and mucocutaneous pigmentation. We present a case report from the Hospital General of Mexico. CLINICAL CASE: We present the case of a 28-year-old male. During physical examination we noted hyperpigmented dermatosis of the oral mucosa and lips. The same condition was seen in both palms. The condition evolved with intolerance to oral feeding and progressive obstructive jaundice. Panendoscopy reported pangastric sessile polyps, as well as being pylorus passable. In the second duodenal portion occupying the region of the ampoule of Vater was a sessile polyp that deformed the region. Exit of bile was not observed through the ampoule. Ultrasound and computed tomography of the abdomen corroborated dilatation of the extrahepatic biliary tract. Two endoprostheses were placed in the bile duct by endoscopic cholangiography, with improvement of biliary obstruction. Roux-en-Y astrojejunoanastomosis was performed because of obstruction of the duodenum by polyps between the second and third portion. Jejunal enterotomy was necessary because of the presence of intraluminal injury formed by a conglomerate of polyps. The patient had a satisfactory evolution. Pathological study reported hamartomatous polyps. CONCLUSIONS: Duodenal obstruction secondary to biliary tract obstruction is a rare manifestation associated with PJS. In these cases, the treatment of choice is polyp resection using endoscopic and/or surgical approach as well as management of the biliary tract obstruction. PMID- 21631982 TI - Atypical intermediate-grade mediastinal carcinoid. Case presentation. AB - BACKGROUND: Approximately 25% of carcinoid tumors develop in the respiratory system. Neuroendocrine carcinoids represent ~5% of all mediastinal tumors and 1 5% of all intrathoracic neoplasms. They contain numerous neurosecretory granules that synthesize, store and release neurohumoral substances that can induce the carcinoid syndrome. CLINICAL CASE: A 21-year-old male presented with a rapidly progressive paraneoplastic syndrome unleashed by an acute urethritis. Two left mediastinal masses were identified and resected. Postoperative evolution has been uneventful during the first year. CONCLUSIONS: We emphasize the importance of early detection of primary and satellite lesions of these tumors including neurohumoral markers and PET/CT scans as in this case, as well as the participation of a multidisciplinary team. PMID- 21631983 TI - Nephron-sparing surgery for clear cell carcinoma in a solitary functional kidney. Case report and literature review. AB - BACKGROUND: Since the introduction of nephron-sparing surgery, patients with small renal masses and diseases affecting overall renal function have greatly benefited. CLINICAL CASE: We present the case of a 62-year-old male with a 6-cm left renal tumor in the medial parahilar segment with right renal unit functionally excluded, secondary to chronic obstruction due to ureteral lithiasis. Right ureteroscopy was performed with pneumatic lithotripsy. Despite localization and size of the tumor, left nephron-sparing surgery was performed in order to preserve renal function. Follow-up of the patient demonstrates no tumor activity. CONCLUSIONS: Open nephron-sparing surgery has been the standard treatment for small renal masses with peripheral localization; however, in specific cases with deteriorated renal function and tumors >5 cm close to the renal hilum, certain risk must be taken in order to perform this procedure that has proven to be highly effective in preserving renal function. Open nephron sparing surgery represents a highly effective procedure in small renal masses with a nonfunctional contralateral kidney. PMID- 21631984 TI - Gastrointestinal stent. AB - The use of intestinal stent is considered the palliative treatment of choice in patients with severe compromised cardiopulmonary reserve or functional class IV who have blockage in the intestinal lumen by tumor growth. The endoscopist should recognize the indications for prosthesis, which depend on the patient and the lesion. This article reviews the characteristics, indications and results of the prosthesis achieved to date. The use of stents has been reported to achieve a technical success of 94%, although clinical success is variable. In general, less serious complications appear in 27% of the patients and, of these, 17% are due to tumor growth within the prosthesis or in its ends, 5% to migration, pain in 2%, and obstruction of the biliary route in 1%. Serious complications occur in 1% of the patients, among which hemorrhage is included. Mortality is infrequent. We conclude that the use of an enteral prosthesis is effective and safe. It is an accessible method that requires an experienced surgeon. For management of obstruction, it has shown to have good results as a cost-effective palliative option. PMID- 21631985 TI - Pathophysiological implications between chronic inflammation and the development of diabetes and obesity. AB - The different theories about the mechanisms involved in the development of metabolic disease and its complications converge in the presence of an etiologic chronic proinflammatory state. Chronic inflammation is, at present, the central pathophysiological mechanism involved in the genesis of metabolic diseases. The multiple interactions between the immune system, adipose tissue, the vascular wall and the pancreas are the issues addressed in this review, focusing on specific intracellular and molecular aspects that may become new therapeutic targets. These lead to a proinflammatory, prothrombotic state as well as to proapoptotic endothelial damage that allows the development of atherosclerosis and, consequently, cardiovascular disease. The multiple immunopathological processes associated with the etiology and pathophysiology of different chronic diseases is still in the process of being fully elucidated, allowing the development of new therapeutic targets. PMID- 21631986 TI - Portopulmonary hypertension. AB - Portopulmonary hypertension (PPHT) is a respiratory complication of portal hypertension, defined as an increase in mean pulmonary artery pressure (PAP) of > 25 mmHg with an increase in pulmonary vascular resistance of > 240 dyn.s/cm(-5) and a normal pulmonary capillary wedge pressure ( < 15 mmHg), which often occurs in subjects with liver cirrhosis. Histopathological features of PPHT are endothelial and smooth-muscle cell proliferation and fibrosis leading to luminal obstruction in the resistance arteries. The pathogenesis of PPHT may result from an imbalance between vasoconstrictor and vasodilating factors. The most common pulmonary symptom is exertional dyspnea; fatigue, chest pain and syncope occur more often at an advanced stage. Edema, ascites and prominent jugular veins are signs of both decompensated hepatic cirrhosis and right ventricular failure. Right heart catheterisation is the gold standard for the diagnosis and defines PPHT in mild disease with PAP less than 35 mmHg, moderate disease with PAP between 35 and 45 mmHg, and severe disease with PAP of 45 mmHg or higher. The medical treatment of portopulmonary hypertension is based on the treatment of other forms of pulmonary arterial hypertension, including vasomodulating pharmacologic agents. Liver transplantation is accompanied by high risk of mortality, generally due to acute right ventricular failure and cardiovascular collapse. The prognosis of PPHT is poor with mean survival of 15 months. PMID- 21631987 TI - Molecular aspects of chronic radiation enteritis. AB - PURPOSE: Chronic radiation enteritis (CRE) is one of the most feared complications of abdominal or pelvic radiation therapy and the treatment of CRE is difficult and often controversial. Recent progress in molecular biology has shed some light on the pathogenesis of CRE, which is characterized by fibrosis. The purpose of this article is to summarize the current state of knowledge of molecular aspects of radiation induced intestinal fibrosis and to discuss potential therapeutic targets. METHODS: A review of the up-to-date published literature involving the possible molecular cascades in radiation-induced intestinal fibrosis and prospective targets for CRE were performed using the Pub Med search engine. RESULTS: Fibrosis development is correlated with transforming growth factor beta1 (TGF-beta1) and its downstream effector Smad3, which stimulates fibrogenic downstream mediators, such as connective tissue growth factor (CTGF). Ras homologue (Rho) and Rho-associated kinase (ROCK) signaling pathway have been shown to play important roles in the development of CRE. The inhibition of these pathways ameliorated radiation-induced intestinal fibrosis in vitro and in animal studies; however, the relationship between the Smad3 and Rho signaling pathways has not been elucidated. CONCLUSIONS: Rho/ROCK and TGF beta1/Smad3 signaling pathways have been shown to play a key role in intestinal fibrogenesis, which might provide with effective possibilities for clinical intervention. Understanding the cooperation between Smad3 and Rho, may therefore be critical to our overall understanding of fibrosis development and maintenance of CRE. PMID- 21631988 TI - The relationship between adiponectin and blood pressure in premenopausal and postmenopausal women. AB - PURPOSE: Menopause can affect the reportedly inverse association between adiponectin and blood pressure (BP); however, this relationship is still poorly understood. The present study cross-sectionally compared the relationship between adiponectin and BP in pre- and postmenopausal women. METHODS: Healthy, asymptomatic women on no medication (n = 262) were divided into a premenopausal group (n = 125, mean age 44.7 years) and a postmenopausal group (n = 137, mean age 65.6 years). Fasting values of serum adiponectin and BP were measured, in addition to body mass index (BMI), blood glucose and lipids. The correlation between the levels of adiponectin/BMI and mean BP (MBP) was analyzed with a linear regression model for the respective groups. RESULTS: The median adiponectin/BMI did not significantly differ between the pre- and postmenopausal groups (0.37 and 0.42, P = 0.08), and the premenopausal group had a significantly lower level of mean MBP than the postmenopausal group (87.6 and 100.7 mmHg, P < 0.001). In an unadjusted analysis, adiponectin/BMI was found to be significantly and inversely correlated with MBP in the premenopausal group (r = -0.499, P < 0.001) and the postmenopausal group (r = -0.203, P < 0.01), respectively. In a stepwise multivariate-adjusted analysis, adiponectin/BMI remained significantly, inversely and independently correlated with MBP in the premenopausal group (beta = -0.383, P < 0.001), while no significant correlation was found between adiponectin/BMI and MBP for the postmenopausal group. CONCLUSIONS: The adiponectin-BP relationship appears to be associated with premenopausal state. PMID- 21631989 TI - Association of serum cortisol levels with parameters of metabolic syndrome in men and women. AB - PURPOSE: Central fat distribution is significantly associated with an increased cortisol levels. We hypothesized that men with metabolic syndrome have a higher serum cortisol levels compared to women. The purpose of this study was to compare serum cortisol levels, and its correlation with serum leptin levels, between men and women with and without metabolic syndrome. METHODS: A cross sectional study was performed with 120 untreated patients with metabolic syndrome and 165 healthy volunteers as controls. Serum lipid profile, fasting blood sugar, insulin, cortisol and leptin levels were measured for both groups. RESULTS: Men with metabolic syndrome had a higher serum cortisol levels, while serum leptin levels were significantly higher in women. The higher serum cortisol level in men with metabolic syndrome was significant after multiple adjustments for age, BMI and waist circumference (17.74+/-5.1 vs 14.07+/-4.3; p < 0.05) using general linear model; however, these difference were no longer significant when the waist-to-hip ratio was added as one of the adjustment factors (16.7+/-1.2 vs. 14.9+/-0.5; p < 0.2). Serum cortisol levels was significantly correlated with serum leptin (r=0.33, p < 0.05), cholesterol (r=0.35, p < 0.05), triglyceride (r=0.25, p < 0.05), waist circumference (r=0.41, p < 0.01) and waist-to-hip ratio (r=0.32, p < 0.01) in women with metabolic syndrome, after controlling for age and BMI. CONCLUSION: Serum cortisol levels are significantly higher in men with metabolic syndrome. This effect is independent of waist circumference. PMID- 21631990 TI - IL-8 reduces VCAM-1 secretion of smooth muscle cells by increasing p-ERK expression when 3-D co-cultured with vascular endothelial cells. AB - OBJECTIVE: The goal of this study was to investigate the crosstalk between vascular endothelial cells (ECs) and smooth muscle cells (SMCs) using a three dimensional (3-D) co-culture model. In addition, the role of IL-8 in this crosstalk was investigated. METHODS: A 3-D co-culture model was constructed using a Transwell chamber system and type I collagen gel. Human umbilical artery smooth muscle cells (HUASMCs) were suspended in the gel and added to the upper compartment of the Transwell. Human umbilical vein endothelial cells (HUVECs) were then grown on the surface of the gel. The growth of HUASMCs was tested with a CFDA SE cell proliferation kit. IL-8 and other bioactive substances were investigated by ELISA and real-time PCR. The alteration of p-ERK expression related to the change in IL-8 levels was also examined by Western blot analysis. RESULTS: The proliferation rate of HUASMCs in the 3-D co-culture model was 0.679 +/- 0.057. Secretion and transcription of VEGF, t-PA, NO and VCAM-1 in the 3-D co culture model were different than in single (2-D) culture. When 3-D co-cultured, IL-8 released by HUVECs was significantly increased (2.35 +/- 0.16 fold) (P<0.05) and the expression of VCAM-1 from HUASMCs was reduced accordingly (0.55+/-0.09 fold). In addition, increasing or decreasing the level of IL-8 changed the level of p-ERK and VCAM-1 expression. The reduction of VCAM-1, resulting from increased IL-8, could be blocked by the MEK inhibitor, PD98059. CONCLUSION: Crosstalk between HUVECs and HUASMCs occurred and was probably mediated by IL-8 in this 3-D co-culture model. PMID- 21631991 TI - Vitamin D and hypertension in pregnancy. AB - PURPOSE: Vitamin D Deficiency is common, particularly in northern latitudes. We examined the association between vitamin D status and hypertension in late pregnancy. METHODS: A case-control study was conducted during two time periods: September-October, 2008, and January-March, 2009, in women near term. A case was defined as having two or more documented blood pressure readings above 140/90 (either/or) at any time during pregnancy (n=78). Controls had at least two blood pressure readings, with none above 140/90 during pregnancy (n=109). Serum 25 hydroxyvitamin D (25(OH)D) was measured in all participants. RESULTS: In the summer, 13% of controls and 29% of the cases had 25(OH) D levels < 50 nmol/L. During the winter, these numbers rose to 44% and 49% respectively. Both cases and controls were more likely to be vitamin D deficient in the winter (p=0.002). There was a negative correlation between BMI and 25(OH)D (r=-0.202, p=0.002). In univariate analysis, cases had lower 25(OH)D (p=0.046), but also higher body mass index, so that in multivariate analysis 25(OH)D status was no longer significant. There was no difference in mean oral daily vitamin D intake (dietary intake and supplements, 746 and 785 IU respectively). Controls gained less weight in pregnancy. There was a negative correlation between the highest blood pressure measured in pregnancy and 25(OH)D levels (r= -0.118; p=0.012). CONCLUSION: There is a high prevalence of vitamin D deficiency in pregnant women recruited in Saskatoon, Saskatchewan. Women with low circulating vitamin D concentrations are more likely to have hypertension. PMID- 21631992 TI - Knockdown of Mgat5 inhibits CD133+ human pulmonary adenocarcinoma cell growth in vitro and in vivo. AB - PURPOSE: In spite of many therapeutic advances, the prognosis of lung cancer remains poor. Therefore, understanding the molecular mechanisms underlying cancer progression, invasion and metastasis is needed. Accumulating evidence indicate that N-acetylglucosaminyltransferase V (Mgat5 or GnT-V) is involved in cancer development. The purpose of this study was to characterize the expression and function of Mgat5 in CD133+ pulmonary adenocarcinoma cells. METHODS: CD133+ pulmonary adenocarcinoma cells were separated by magnetic activated cell sorting (MACS) from excised pulmonary adenocarcinoma specimens from 10 patients. Expression of Mgat5 in CD133+ cells was detected by fluorescent quantitative RT PCR (FQRT-PCR) and Western blot. Subsequently, CD133+ cells were transfected with specific siRNA of Mgat5 to evaluate the effects of Mgat5 inhibition on cancer cell growth in vivo and in vitro. RESULTS: Expression of Mgat5 was 1.2-fold and 1.4-fold higher in CD133+cells than in CD133- cells detected by FQRT-PCR and Western Blot, respectively (p < 0.05). The L-PHA binding assay also showed higher reactivity in CD133+ cells than in CD133- cells. In addition, Mgat5-specific siRNA efficiently knocked down the expression of Mgat5 in CD133+ cells. Interestingly, downregulation of Mgat5 resulted in significant inhibition of cancer cell growth in vitro and in vivo. CONCLUSION: Mgat5 is expressed at a relatively high level in CD133+ lung adenocarcinoma cells, and knockdown of Mgat5 in CD133+ cells inhibits cancer cell growth both in vitro and in vivo. These findings suggest Mgat5 may play an important role during oncogenesis, identifying a potential therapeutic target for pulmonary adenocarcinoma. PMID- 21631993 TI - Oxidative lipid, protein, and DNA damage as oxidative stress markers in vascular complications of diabetes mellitus. AB - PURPOSE: The purpose of this study was to determine the effects of diabetic complications on oxidation of proteins, lipids, and DNA and to investigate the relationship between oxidative damage markers and clinical parameters. METHODS: The study group consisted of 69 type 2 diabetic patients (20 patients without complication, 49 patients with complication) who attended internal medicine outpatient clinics of Istanbul Education and Research Hospital and 19 healthy control subjects. In serum samples of both diabetic patients and healthy subjects, 8-hydroxy-2'deoxyguanosine (8-OHdG), as a marker of oxidative DNA damage, N(epsilon)-(hexanoyl)lysine (HEL) and 15-F2t-iso-prostaglandin (15-F2t IsoP). as products of lipooxidative damage, advanced oxidation protein products (AOPP), as markers of protein damage, and paraoxonase1 (PON1) as antioxidant were studied. RESULTS: 15-F2t-IsoP (p < 0.005) and AOPP (p < 0.001) levels were significantly higher in diabetic group than control group while there were no significant differences in levels of 8-OHdG and HEL between the two groups. AOPP (p < 0.001) and 8-OHdG (p < 0.001) were significantly higher in diabetic group with complications compared to diabetic group without complications. CONCLUSIONS: Increased formation of free radicals and oxidative stress, under conditions of hyperglycaemia, is one of the probable causes for evolution of complications in diabetes mellitus. Our study supports the hypothesis that oxidant/antioxidant balance is disturbed in diabetic patients. PMID- 21631994 TI - Increased NPC1L1 and serum cholesterol in a chronic rejection rat. AB - PURPOSE: To measure serum cholesterol and triglyceride levels and NPC1L1 mRNA and protein as an index of cholesterol absorption during the development of chronic rejection (CR) in a rat model of intestinal transplantation. METHODS: Rats were randomly divided into two groups: Group 1 (n=20) underwent syngenic Lewis-to Lewis transplantation and Group 2 (n=20) underwent allogenic F344-to-Lewis transplantation as well as treatment with FK506. Blood samples and intestinal tissue were procured on the 190th day after operation. Histological changes were analyzed and the semiquantitative scores of histological parameters were compared. The serum levels of cholesterol and triglyceride were determined. The expression of Niemann-Pick C1 Like 1(NPC1L1) mRNA and protein were analyzed by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) and immunohistochemistry, respectively. RESULTS: All the animals survived for the 190 days. The appearance and histology of isografts were almost normal whereas the allografts displayed thickened bowel wall and mesenteric fibrosis, concentric intimal thickening and interstitial fibrosis and inflammatory infiltration. The histology scores displayed a significant difference between the allografts and isografts (P < 0.001). No differences were observed for triglycerides for the two groups. The serum cholesterol levels increased significantly in the allogenic group in comparison with the syngenic group (P=0.034), while no difference was observed for triglyceride levels between groups. RT-PCR showed that the expression of NPC1L1 of allografts increased significantly (P=0.004). Immunohistochemistry confirmed RT-PCR findings. CONCLUSIONS: Neointima formation and mesenteric fibrosis were the dominant pathological features. The increased expression of NPC1L1 might contribute to hypercholesterolemia, which may be involved in the pathogenesis of transplant arteriosclerosis. PMID- 21631995 TI - Assessment of anti-sperm antibodies in couples after testicular sperm extraction. AB - PURPOSE: Testicular spermatozoa can be retrieved successfully by the testicular sperm extraction (TESE) procedure and used for intracytoplasmic sperm injection. Disruption in the blood-testis barrier can lead to the production of antisperm antibodies (ASA). The aim of this prospective study was to investigate the frequency of ASA formation in couples after TESE procedure. METHODS: Thirty-seven couples were included in the study at the Urology Clinic of the Dr. Zekai Tahir Burak Women's Health Training and Research Hospital. History, physical examination, spermiogram, and endocrine profiles were obtained for all male patients. All the male patients in this study had been diagnosed with nonobstructive azoospermia (NOA) and underwent microdissection TESE. Secondary and tertiary cases were also included in the study. Serum samples were obtained from all 74 patients before TESE, and at three and 12 months after TESE. Serum ASA levels were determined. ANOVA was performed for statistical analysis for serum Follicle-Stimulating Hormone (FSH), testosterone and testicular volume. P < 0.05 was considered significant. RESULTS: There were no differences in the testicular volumes, serum FSH and testosterone levels before and after TESE. None of the patients or their partners developed significant levels of ASA as a result of the TESE procedure. CONCLUSION: TESE procedure does not cause ASA production in either males or their female partners. PMID- 21631996 TI - Reduced expression of DeltaNup63alpha in cervical squamous cell carcinoma. AB - PURPOSE: As a member of the p53 family, p63 is considered to be an important differentiation regulation transcriptional factor, but the roles of p63 in many epithelial tumourigenesis and metastasis processes are still not clear. This study was designed to investigate the expression of p63 and its isoform in normal tissues and squamous cell cancer tissues of uterine cervix, and its significance in cancer cell differentiation. METHODS: The expression of p63 was assessed in cervical tissue and cell lines by immunohistochemistry, RT-PCR and Western Blotting. The relationships between p63 protein, various clinico-pathological features, and the differentiation marker involucrin were analyzed. RESULTS: DeltaNup63alpha is the predominant isoform expressed in cervical epithelial tissues, and it is decreased in moderately or poorly differentiated cervical squamous carcinoma, as well as in the HeLa, SiHa and C33A cervical cancer cell lines. The expression level of DeltaNup63alpha was positively correlated with that of involucrin in cervical squamous cancer tissue, and the expression of DeltaNup63alpha is decreased with the degree of tumour invasion. CONCLUSION: The decrease of DeltaNup63alpha in cervical squamous cell cancer appears to be associated with the tumour progression, and DeltaNup63alpha may be a sensitive marker for cervical squamous cancer differentiation. PMID- 21631997 TI - Falls prevention in NSW: a big issue requiring sustained research, policy and practice initiatives. PMID- 21631998 TI - Prevalence, circumstances and consequences of falls among community-dwelling older people: results of the 2009 NSW Falls Prevention Baseline Survey. AB - AIM: To describe the prevalence, circumstances and consequences of falls among community-dwelling older people in NSW using data from the 2009 NSW Falls Prevention Baseline Survey. METHODS: Telephone interviews with a random sample of 5681 NSW residents aged 65 years and over were conducted in 2009. RESULTS: Of those surveyed, 25.6% reported falling in the last year. Of those who fell, 61.2% fell once, 21.4% fell twice, 7.8% fell three times, and 9.5% fell four or more times in the last year. Sixty-six percent of those who fell in the last year were injured and 20.0% visited a hospital as a result of a fall. The most common injuries were cuts, grazes or bruises (71.0%) and sprains or strains (9.9%). CONCLUSION: The findings of this survey are consistent with previous findings in the published fall injury prevention literature. The results from the survey will assist in the design of community oriented fall injury prevention strategies and will form the baseline measure for the evaluation of the impact of these strategies in NSW. PMID- 21631999 TI - Characteristics of fall-related injuries attended by an ambulance in Sydney, Australia: a surveillance summary. AB - In NSW, fall-related injury costs the health system more than any other single cause of injury. A public health surveillance database containing information routinely recorded by the Ambulance Service of NSW was used to define the epidemiology and characteristics of fall-related calls in the Sydney metropolitan area in 2008. The dataset contained 37488 fall-related calls, representing a crude rate of ambulance call-outs for falls of 843 per 100000 population. Females accounted for 57% of all fall-related calls, and the female rate of injury to the 'hip to foot' region increased with age. Males in all age groups reported 'head and neck' injury most often. In an analysis of a random sample of 1200 calls, 70% of ambulance dispatches were to a home or residential institution. The findings of this study on the risks for fall-related injury can be used to guide policy for ambulance service delivery. Expansion of data linkage to emergency department and admitted patient databases would provide information to further describe the epidemiology of falls in NSW. PMID- 21632000 TI - The cost of fall-related injuries among older people in NSW, 2006-07. AB - This study aimed to establish comprehensive estimates of the cost of fall-related injury among older people in NSW. A health service utilisation approach was used to estimate the cost of hospital treatment, residential care and ambulance transport. Other costs were estimated by deriving ratios of inpatient costs to other services from the literature. In the 2006-07 financial year, 251,000 (27%) of older people fell at least once and suffered, in total, an estimated 507,000 falls. An estimated 143000 medically treated fall-related injuries among older people resulted in lifetime treatment costs of $558.5 million. Although only 18% of these injuries resulted in hospital admission, the cost of care associated with these cases accounted for 84.5% of the total cost. The cost of fall-related injury among older people in NSW in 2006-07 is a significant increase over earlier estimates and underscores the urgent need for effective preventive efforts across the state. PMID- 21632001 TI - An economic evaluation of community and residential aged care falls prevention strategies in NSW. AB - AIM: To evaluate the cost-effectiveness of strategies designed to prevent falls amongst people aged 65 years and over living in the community and in residential aged-care facilities. METHODS: A systematic review and meta-analysis of the literature was conducted. The pooled fall rate ratio was used in a decision analytic model that combined a Markov model and decision tree to estimate the costs and outcomes of potential interventions and/or strategies. The resulting cost per quality-adjusted life year was estimated. RESULTS: The most cost effective falls prevention strategy in community-dwelling older people was Tai Chi. Expedited cataract surgery and psychotropic medication withdrawal were also found to be cost-effective; however, the effectiveness of these interventions is less certain due to small numbers of trials and participants. The most cost effective falls prevention strategies in residential aged-care facilities were medication review and vitamin D supplementation. PMID- 21632002 TI - Evaluation of the NSW Management Policy to Reduce Fall Injury Among Older People, 2003-2007: implications for policy development. AB - AIM: To retrospectively evaluate the NSW Management Policy to Reduce Fall Injury Among Older People, 2003-2007. METHODS: The process evaluation was conducted by interviewing stakeholders regarding the implementation of the Policy. A document review was also undertaken to identify activities and initiatives undertaken during the implementation process. RESULTS: Key achievements in the implementation of the Policy and significant early events, decisions and contextual factors which acted as barriers to the implementation were identified. Results included the: identification of the need for the prioritisation of evidence-based interventions; application of consistent best practice guidelines for implementing falls prevention strategies in the community; and development of an evaluation and monitoring framework concurrent with the development of policy. CONCLUSION: Subsequent policy initiatives must be strategically focused and coordinated if future activities are to have a significant impact on the increasing burden of fall-related injury. PMID- 21632003 TI - The strategic development of the NSW Health Plan for Prevention of Falls and Harm from Falls Among Older People: 2011-2015; translating research into policy and practice. AB - With our rapidly ageing population there is an urgent imperative to minimise the rate of falls and associated injuries. A key challenge to public health is to better conceptualise and contextualise falls prevention evidence for more effective policy making and practice. This paper describes how NSW Health adopted the Nutbeam and Bauman Stages of Research and Evaluation Model in the strategic development of the NSW Health Plan for Prevention of Falls and Harm from Falls Among Older People: 2011-2015. Research evidence has been comprehensively applied to every stage of the development of the Plan and research and evaluation is a key action area within the new Plan. The Stages of Research and Evaluation Model provides a useful overarching framework for policy makers to contextualise and more effectively apply research evidence throughout the policy making process from problem definition to program monitoring. PMID- 21632004 TI - Exercise to prevent falls in older adults: an updated meta-analysis and best practice recommendations. AB - This systematic review update includes 54 randomised controlled trials and confirms that exercise as a single intervention can prevent falls (pooled rate ratio 0.84, 95% CI 0.77-0.91). Meta-regression revealed programs that included balance training, contained a higher dose of exercise and did not include walking training to have the greatest effect on reducing falls. We therefore recommend that exercise for falls prevention should provide a moderate or high challenge to balance and be undertaken for at least 2 hours per week on an ongoing basis. Additionally, we recommend that: falls prevention exercise should target both the general community and those at high risk for falls; exercise may be undertaken in a group or home-based setting; strength and walking training may be included in addition to balance training but high risk individuals should not be prescribed brisk walking programs; and other health-related risk factors should also be addressed. PMID- 21632005 TI - Implementing falls prevention research into policy and practice: an overview of a new National Health and Medical Research Council Partnership Grant. AB - Preventing falls and fall-related injuries among older people is an urgent public health challenge. This paper provides an overview of the background to and research planned for a 5-year National Health and Medical Research Council Partnership Grant on implementing falls prevention research findings into policy and practice. This program represents a partnership between key Australian falls prevention researchers, policy makers and information technology companies which aims to: (1) fill gaps in evidence relating to the prevention of falls in older people, involving new research studies of risk factor assessment and interventions for falls prevention; (2) translate evidence into policy and practice, examining the usefulness of new risk-identification tools in clinical practice; and (3) disseminate evidence to health professionals working with older people, via presentations, new evidence-based guidelines, improved resources and learning tools, to improve the workforce capacity to prevent falls and associated injuries in the future. PMID- 21632006 TI - Bug breakfast in the bulletin: rotavirus. PMID- 21632007 TI - Communicable diseases report, NSW, January and February 2011. PMID- 21632017 TI - Coverage of human papillomavirus vaccination during the first year of its introduction in Spain. AB - The decision to introduce human papillomavirus (HPV) vaccination into the national immunisation programme in Spain was made in October 2007, recommending vaccination of girls aged between 11 and 14 years with three doses of HPV vaccine. All 19 regions of the country (17 Autonomous Communities and two Autonomous Cities) introduced HPV vaccination gradually into their immunisation programmes between November 2007 and the last school term of 2008. Eight regions administered the vaccine in healthcare centres and 11 in schools. In the first year of the introduction of HPV vaccination, coverage of the first and third doses was assessed, to determine the proportion of girls who did not complete the vaccination. On the basis of the available data, the Ministry of Health estimated that coverage for the first dose was 87.2% (range: 73.9-98.9%; 95% CI: 71.8 to 100) and 77.3% (range: 62.2-97.4%; 95% CI: 57.9 to 96.7) for the third dose. Higher uptake was observed when the vaccination was carried out in schools compared with healthcare centres, but the difference was not statistically significant. Negative messages in the media during implementation of the HPV vaccination programme may have had some influence on the attitudes of adolescent girls and/or their parents towards HPV vaccination and may be partly responsible for the observed vaccination dropout rate. PMID- 21632018 TI - Outbreak of hand, foot and mouth disease caused by Coxsackie A16 virus in a childcare centre in Croatia, February to March 2011. AB - We describe an outbreak of hand, foot and mouth disease (HFMD) in a childcare centre in a district of Zagreb county, north-west Croatia. A total of eleven cases of HFMD occurred in the childcare centre and another nine were reported from nearby areas in the district. Coxsackie A16 virus was diagnosed in 13 clinical specimens obtained from 11 symptomatic and asymptomatic children. All cases resolved without complications. PMID- 21632019 TI - False-negative results using Neisseria gonorrhoeae porA pseudogene PCR - a clinical gonococcal isolate with an N. meningitidis porA sequence, Australia, March 2011. AB - The gonococcal porA pseudogene is a popular target for in-house Neisseria gonorrhoeae PCR methods. With this study we present two novel findings: the first case of an N. gonorrhoeae porA pseudogene PCR false-negative result caused by sequence variation, and in the same organism, the first description of a clinical N. gonorrhoeae strain harbouring an N. meningitidis porA sequence. PMID- 21632021 TI - Waterborne nickel bioaccumulation in Gammarus pulex: comparison of mechanistic models and influence of water cationic composition. AB - The biodynamic and saturation models offer promising lines of enquiry to predict the bioaccumulation of metals by aquatic organisms. However, in order to construct these models, the accumulation strategies have to be defined for each metal/organism couple in controlled conditions. This study aims at modelling the waterborne bioaccumulation of Ni and the influence of the water's geochemical properties on this process in a crustacean that is widely distributed in Europe, Gammarus pulex. In the laboratory, G. pulex was exposed to several Ni concentrations (from 0.001 to 100 mg L(-1)) in aquatic microcosms. Our results show that G. pulex is very tolerant to Ni (LC50(48 h)=477 mg L(-1) Ni). Time course experiments enabled the construction of a biodynamic model by determining the uptake (k(u)) and elimination (k(e)) rate constants. When the exposure concentration exceeded 1 mg L(-1) Ni, the metal uptake reached a maximum due to a limited number of binding sites for Ni. Therefore, the organism's maximal capacity to accumulate the metal (B(max)) and the half-saturation constant (K) were determined to establish the saturation model. We showed that the two models are comparable for the lowest exposure concentrations (<1 mg L(-1) Ni), with k(u)/k(e)=B(max)/K. Then, the bioaccumulation of Ni was recorded in waters exhibiting various concentrations of three major ions (Na(+), Mg(2+) and Ca(2+)). Only Ca had an inhibitory effect on the Ni uptake. This study reports for the first time the bioaccumulation of Ni in G. pulex. Because of its high tolerance to Ni and its high capacity to accumulate this metal, this crustacean could be used as an indicator of Ni bioavailability in freshwaters. PMID- 21632020 TI - Large and ongoing outbreak of haemolytic uraemic syndrome, Germany, May 2011. AB - Since early May 2011, an increased incidence of haemolytic uraemic syndrome (HUS) and bloody diarrhoea related to infections with Shiga toxin-producing Escherichia coli (STEC) has been observed in Germany, with most cases in the north of the country. Cases reported from other European countries had travelled to this area. First results of a case-control study conducted in Hamburg suggest an association between the occurrence of disease and the consumption of raw tomatoes, cucumber and leaf salad. PMID- 21632022 TI - Unravelling the pathway of respiratory toxicity in goldlined seabream (Rhabdosargus sarba) induced by the harmful alga Chattonella marina. AB - The harmful alga Chattonella marina has caused massive fish kills and economic losses worldwide. Suffocation is generally believed to be the major cause of fish death by C. marina. However, the specific mechanisms leading to respiratory disorder in fish and subsequent fish kills by C. marina remain unknown. The goldlined seabream, highly susceptible to C. marina, was employed to investigate temporal changes of physiological, histopathological and biochemical parameters related to respiratory function at different stages of exposure to C. marina. Hemoglobin oxidation and blood lyses were not found in goldlined seabream exposed to C. marina, which could not be the key reasons accounting for pO(2) drop in the stressed fish. Gill histopathology such as irregular organization of lamellae, mucous with algal cells trapped in interfilamental spaces, were typical in C. marina exposed fish. A surge of plasma lactate occurred in goldlined seabream shortly after exposure to C. marina (0.5h) and sustained throughout the exposure period, indicating rapid onset of and persistent anaerobic respiration in C. marina exposed fish. Depletion of plasma glucose was clearly evident in goldlined seabream showing stress symptoms and near death. Yet, fish alive in the C. marina bloom did not exhibit plasma glucose depletion. The results suggest that availability of fermentable fuel as indicated by glucose level is critical to determine fish survival in C. marina exposure. Overall, our findings have rebuked the involvement of hemolysins and/or nitric oxide as the culprits for C. marina toxicity to fish. This study is the first to demonstrate the pathway of respiratory toxicity induced by the harmful alga C. marina in fish. PMID- 21632023 TI - Transcriptional responses in neonate and adult Daphnia magna in relation to relative susceptibility to genotoxicants. AB - Little information is available on the responses of lower animals to genotoxic chemicals or on their sensitivity for detecting genotoxic chemicals, especially at different life-stages, despite the established use of the water flea Daphnia magna in ecotoxicity testing. Comet assay methodology was developed and applied to daphnid cells but only limited, non-statistically significant responses to the genotoxicants sodium dichromate (0.2-1 MUM), chrysoidine (0.1-2 MUM), and mixtures of benzo-a-pyrene (BaP) and sodium dichromate were found (from 0.01 MUM BaP & 0.1 MUM sodium dichromate to 0.25 MUM BaP & 0.75 MUM sodium dichromate). Transcriptomic analyses using Agilent D. magna oligonucleotide microarrays were undertaken to assess the effect of a mixture of sodium dichromate and BaP (designed to produce both adducted and oxidised DNA) on gene transcription. Neonates (<24h) and adults (day 7) were exposed for 6h and 24h at two combination concentration levels (0.02 MUM BaP & 0.15 MUM sodium dichromate and 0.1 MUM BaP & 0.75 MUM sodium dichromate). The greatest differences in transcriptional profile occurred between adults and neonates. Subsets of the transcriptional profiles distinguished genotoxicant-exposed animals from controls, both for neonates and adults. Higher transcript levels of DNA repair genes were found in adults and adults also displayed significant induction of DNA repair gene transcripts in response to exposure whereas neonates did not. Transcriptional changes in response to genotoxicant exposure proved more sensitive than measurement of DNA strand breaks by the Comet assay and the extensive differences in transcription between adults and neonates emphasized the importance of life stage in toxicant testing with Daphnia. PMID- 21632024 TI - The mechanism of anthracene interaction with photosynthetic apparatus: a study using intact cells, thylakoid membranes and PS II complexes isolated from Chlamydomonas reinhardtii. AB - Intact cells of Chlamydomonas reinhardtii as well as isolated thylakoid membranes and photosystem II complexes were used to examine a possible mechanism of anthracene (ANT) interaction with the photosynthetic apparatus. Since ANT concentrations above 1 mM were required to significantly inhibit the rate of oxygen evolution in PS II membrane fragments it may indicate that the toxicant did not directly interact with this photosystem. On the other hand, stimulation of oxygen uptake by ANT-treated thylakoids suggested that ANT could either act as an artificial electron acceptor in the photosynthetic electron transport chain or function as an uncoupler. Electron transfer from excited chlorophyll to ANT is impossible due to the very low reduction potential of ANT and therefore we propose that toxic concentrations of ANT increase the thylakoid membrane permeability and thereby function as an uncoupler, enhancing electron transport in vitro. Hence, its unspecific interference with photosynthetic membranes in vitro suggests that the inhibitory effect observed on intact cell photosynthesis is caused by uncoupling of phosphorylation. PMID- 21632025 TI - Proteomic modification in gills and brains of medaka fish (Oryzias melastigma) after exposure to a sodium channel activator neurotoxin, brevetoxin-1. AB - Although brevetoxins (PbTxs) produced by the marine dinoflagellate Karenia brevis are known to be absorbed across gill membranes and exert their acute toxic effects through an ion-channel mediated pathway in neural tissue, the exact biochemical mechanism concerning PbTxs neurotoxicity in neural tissue and gas exchange organs has not been well elucidated. In this study, we calculated the LC(50) value of PbTx-1 using the medaka fish model, and presented the molecular responses of sub-acute exposure to PbTx-1 with proteomic method. By adopting two dimensional electrophoresis, the abundances of 14 and 24 proteins were found to be remarkably altered in the gills and brains, respectively, in response to toxin exposure. Thirteen gill and twenty brain proteins were identified using matrix assisted laser desorption/ionization tandem time-of-flight mass spectrometry analysis. These proteins could be categorized into diverse functional classes such as cell structure, macromolecule metabolism, signal transduction and neurotransmitter release. These findings can help to elucidate the possible pathways by which aquatic toxins affect marine organisms within target organs. PMID- 21632026 TI - Bisphenol A modulates expression of sex differentiation genes in the self fertilizing fish, Kryptolebias marmoratus. AB - Endocrine disrupting chemicals (EDCs) have been a major concern in the normal reproduction and development of aquatic organisms. In the teleost, steroid hormones are synthesized via the steroidogenesis pathway, and play a key physiological role in the regulation of gonadal sex differentiation. The protogynous hermaphroditic fish, Kryptolebias marmoratus is the only vertebrate capable of reproducing through internal self-fertilization. To uncover the effect of bisphenol A (BPA) on sex differentiation genes on transcription, we investigated the expression patterns of several sex differentiation-related genes such as dax1, dmrt1, mis, sf1, figlalpha, StAR and wt1 after BPA exposure with controls (E2 and TMX). In response to 17beta-estradiol (E2) exposure, a testis specific gene, dmrt1 mRNA was down-regulated in the gonad of the secondary male but the expression of the female-specific gene, dax1 mRNA was significantly elevated in the brain and gonad. A high level of StAR mRNA was detected in the brain and gonad of both hermaphrodite and secondary males, suggesting that the elevated expression of dax1 and StAR genes would be involved in E2 exposure. As expected, upon BPA exposure, the dmrt1 and MIS mRNA level decreased in both hermaphrodite and secondary males, while the female-specific gene, figlalpha mRNA level increased in the gonad of both genders. BPA showed an opposite mode of action on the expression of dax1 (induction, P>0.05) and sf1 mRNA (inhibition, P>0.05) in the brain and gonad against both genders. The sensitivity of dax1 to BPA on expression was relatively high in the secondary male. The wt1 mRNA was up regulated in most tissues except in the liver of BPA-exposed secondary males. Regarding the time course study, the figlalpha mRNA level increased at 6 h after BPA exposure. In addition, BPA elevated the expression of StAR, dax1, and wt1 mRNA but repressed sf1 mRNA. In this paper, we demonstrated that BPA may modulate the expression of sex differentiation and steroidogenesis pathway genes, and this finding would provide a better understanding on the modulation of transcription upon BPA exposure in steroidogenesis and sex differentiation in the hermaphroditic fish, K. marmoratus. PMID- 21632027 TI - Exposure to atrazine affects the expression of key genes in metabolic pathways integral to energy homeostasis in Xenopus laevis tadpoles. AB - In our laboratory, Xenopus laevis tadpoles exposed throughout development to 200 or 400 MUg/L atrazine, concentrations reported to periodically occur in puddles, vernal ponds and runoff soon after application, were smaller and had smaller fat bodies (the tadpole's lipid storage organ) than controls. It was hypothesized that these changes were due to atrazine-related perturbations of energy homeostasis. To investigate this hypothesis, selected metabolic responses to exposure at the transcriptional and biochemical levels in atrazine-exposed tadpoles were measured. DNA microarray technology was used to determine which metabolic pathways were affected after developmental exposure to 400 MUg/L atrazine. From these data, genes representative of the affected pathways were selected for assay using quantitative real time polymerase chain reaction (qRT PCR) to measure changes in expression during a 2-week exposure to 400 MUg/L. Finally, ATP levels were measured from tadpoles both early in and at termination of exposure to 200 and 400 MUg/L. Microarray analysis revealed significant differential gene expression in metabolic pathways involved with energy homeostasis. Pathways with increased transcription were associated with the conversion of lipids and proteins into energy. Pathways with decreased transcription were associated with carbohydrate metabolism, fat storage, and protein synthesis. Using qRT-PCR, changes in gene expression indicative of an early stress response to atrazine were noted. Exposed tadpoles had significant decreases in acyl-CoA dehydrogenase (AD) and glucocorticoid receptor protein (GR) mRNA after 24 h of exposure, and near-significant (p=0.07) increases in peroxisome proliferator-activated receptor beta (PPAR-beta) mRNA by 72 h. Decreases in AD suggested decreases in fatty acid beta-oxidation while decreases in GR may have been a receptor desensitization response to a glucocorticoid surge. Involvement of PPAR-beta, an energy homeostasis regulatory molecule, also suggested changes in energy status. Despite, or possibly because of, these early gene changes, there were no differences in either absolute ATP levels or ADP:ATP ratios early in the exposure. However, livers from animals exposed to 200 MUg/L atrazine had near-significant (p=0.06) increases in ADP:ATP ratios at the end of exposure suggesting tadpoles may have had difficulty maintaining energy homeostasis. Perturbations in the expression of genes regulating energy metabolism by 24 h into exposure to 400 MUg/L atrazine was noteworthy, especially since these tadpoles were significantly smaller than controls by 72 h of exposure. PMID- 21632028 TI - Activation of the P2Y1 receptor induces apoptosis and inhibits proliferation of prostate cancer cells. AB - G protein-coupled receptors, the largest cell surface receptor family, have emerged as critical players in cell death and survival. High gene expression level of the G(q)-coupled P2Y(1) nucleotide receptor in PC-3 prostate cancer cells was demonstrated using real-time quantitative PCR and confirmed by Western blotting and confocal laser scanning microscopy. A selective P2Y(1) receptor agonist, the ADP analogue MRS2365, concentration-dependently induced intracellular calcium mobilization (EC(50) 5.28nM), which was diminished by P2Y(1) receptor-selective antagonist MRS2500. P2Y(1) receptor activation by MRS2365 induced apoptosis in assays of Caspase-3, LDH release, and annexin-V staining. The pro-apoptotic effect of MRS2365 was blocked by MRS2500, P2Y(1) siRNA, and an inhibitor of the MAP kinase pathway PD98059. MRS2365 significantly inhibited the proliferation of PC-3 cells, examined using a MTT assay. Thus, activation of the P2Y(1) receptor induced cell death and inhibited growth of human prostatic carcinoma PC-3 cells. Activation of the P2Y(1) receptor should be a novel and promising therapeutic strategy for prostate cancer. PMID- 21632029 TI - Evolutionary dynamics of the American African genotype of dengue type 1 virus in India (1962-2005). AB - Dengue is a major health problem in India with all four serotypes represented. Recently there has been an increase in the occurrence of dengue-1 outbreaks. It is possible that there have been changes in the genetics of dengue virus-1 (DENV 1), either by fresh introductions or by evolution in situ. The studies on DENV-1 evolution so far have no Indian sequences included. To gain insight into the dynamics of DENV-1 in India, the envelope (E) gene of thirteen virus isolates representative of the period 1962-2005 were sequenced and analyzed together with the available sequences of 40 globally representative isolates. All the Indian DENV-1 isolates were found to belong to the American African (AMAF) genotype. With the addition of 13 Indian isolates, the AMAF genotype can now be called Cosmopolitan. The Indian isolates were distributed into four lineages, India I, II, III and the Africa lineage, now called Afro-India. Of these, India III was the oldest and extinct lineage; the Afro-India was a transient lineage while India I, imported from Singapore and India II, evolving in situ, were the circulating lineages. Despite the extinction and introduction of lineages, no specific codon site was observed to be under selection pressure. The rate of nucleotide substitution estimated for DENV-1 was 6.5 * 10(-4) substitutions/site/year, and the time to the most recent common ancestor (tMRCA) was estimated to be 78-180 years (1825-1925), similar to previous estimates. The tMRCA for the AMAF/Cosmopolitan genotype was 56-98 years (1907-1949), a period that covers World War I and II. The two imports from Africa (1953-1968) and Singapore (1964-1975) and an export to the Americas (1955-1965) prove that there have been changes in the lineage of the DENV-1 viruses circulating in India which has contributed to the global dynamics of DENV-1 evolution and perhaps to the changing epidemiology of dengue in India. PMID- 21632030 TI - Object-based attention: sensory enhancement or scanning prioritization. AB - The three experiments reported here test whether object-modulated attentional spreading can be obtained when the target location is 100% certain. Experiment 1 uses the reaction time (RT)-based flanker task similar to Shomstein and Yantis (2002), and replicates the null result of the object-modulated attentional spreading. RT and accuracy (ACC) have been shown to reflect different processes: postperceptual decision vs. perceptual process (Santee & Egeth, 1982). Experiment 2 adopts the data-limited ACC-based measure and reports that attention could spread within the attended object. To avoid ceiling effects, Experiment 3 adjusts the presentation time based on the trials where the target and flankers were compatible and on the same objects, and provides the convergent evidence supporting the object-modulated attentional spreading. These results suggest that because the RT-based measure is less sensitive in reflecting the quality of perceptual representations, it is not sufficiently a strong evidence to distinguish between sensory enhancement and scanning prioritization accounts. PMID- 21632031 TI - Interparticle interactions in concentrated suspensions and their bulk (rheological) properties. AB - The interparticle interactions in concentrated suspensions are described. Four main types of interactions can be distinguished: (i) "Hard-sphere" interactions whereby repulsive and attractive forces are screened. (ii) "Soft" or electrostatic interactions determined by double layer repulsion. (iii) Steric repulsion produced by interaction between adsorbed or grafted surfactant and polymer layers. (iv)and van der Waals attraction mainly due to London dispersion forces. Combination of these interaction energies results in three main energy distance curves: (i) A DLVO type energy-distance curves produced by combination of double layer repulsion and van der Waals attraction. For a stable suspension the energy-distance curve shows a "barrier" (energy maximum) whose height must exceed 25kT (where k is the Boltzmann constant and T is the absolute temperature). (ii) An energy-distance curve characterized by a shallow attractive minimum at twice the adsorbed layer thickness 2delta and when the interparticle distance h becomes smaller than 2delta the energy shows a sharp increase with further decrease of h and this is the origin of steric stabilization. (iii) an energy-distance curve characterized by a shallow attractive minimum, an energy maximum of the DLVO type and a sharp increase in energy with further decrease of h due to steric repulsion. This is referred to as electrosteric repulsion. The flocculation of electrostatically and sterically stabilized suspensions is briefly described. A section is devoted to charge neutralization by polyelectrolytes and bridging flocculation by polymers. A distinction could be made between "dilute", "concentrated" and "solid suspensions" in terms of the balance between the Brownian motion and interparticle interaction. The states of suspension on standing are described in terms of interaction forces and the effect of gravity. The bulk properties (rheology) of concentrated suspensions are described starting with the case of very dilute suspensions (the Einstein limit with volume fraction Phi<=0.01), moderately concentrated suspensions (0.2>Phi>=0.1) taking into account the hydrodynamic interaction and concentrated suspensions (Phi>0.2) where semi-empirical theories are available. The rheological behavior of the above four main types of interactions is described starting with "hard-sphere" systems where the relative viscosity-volume fraction relationship could be described. The rheology of electrostatically stabilized suspensions was described with particular reference to the effect of electrolyte that controls the double layer extension. The rheology of sterically stabilized systems is described using model polystyrene suspensions with grafter poly(ethylene oxide) layers. Finally the rheology of flocculated suspensions was described and a distinction could be made between weakly and strongly flocculated systems. PMID- 21632032 TI - Factors associated with delays to emergency care for bowel obstruction. AB - BACKGROUND: Our objective was to determine factors associated with delays to first treatment for emergency department (ED) patients diagnosed with small-bowel obstruction (SBO). METHODS: This was a retrospective study of ED patients with SBO. Data were collected from medical records, administrative databases, and staffing schedules at an urban, tertiary care medical center from June 1, 2001, to November 30, 2002. Patient-related characteristics and processes of ED and hospital care were evaluated. Outcomes studied were time to first treatment (nasogastric tube or surgery) and risk of surgical resection. RESULTS: A total of 193 patients were diagnosed with confirmed intestinal obstruction. Patients with longer times to first treatment arrived during ED clinician hand-offs (adjusted hazard ratio, .40; 95% confidence interval, .17-.98). Patients with longer times to surgery consult (ref. first quartile) had greater odds of surgical resection (second quartile adjusted odds ratio, 6.91; 95% confidence interval, 1.85-24.80). CONCLUSIONS: Remediable ED and hospital factors were associated with longer times to treatment for patients with bowel obstruction. PMID- 21632033 TI - Some important oversights in the assessment of whole-body vibration exposure based on ISO-2631-1. PMID- 21632034 TI - Evaluation of occupational injuries in an urban emergency medical services system before and after implementation of electrically powered stretchers. AB - Musculoskeletal injuries are frequently reported among Emergency Medical Services (EMS) professionals. The objective of this study was to evaluate occupational injuries in an urban EMS system before and after implementation of hydraulic stretchers. Data for this analysis were obtained from Austin Travis County EMS (A/TCEMS). In December 2006, A/TCEMS placed into service electrically powered patient stretchers. The pre-intervention period was between 01/01/1999 and 12/31/2006, and the post-intervention period was between 01/01/2007 and 4/30/2008. Incidence rate calculations were performed for four injury sub-groups and rate ratios (RRs) and corresponding 95% confidence interval (CI) were presented. There were 2087 and 706 person-years of observation pre- and post intervention, respectively. The incidence rates for overall injury pre intervention and post-intervention were 61.1 and 28.8 per 100 FTE, with a corresponding RR of 0.47 (95% CI 0.41-0.55) indicating a significant decrease in the rate of injury. The subcategory of stretcher-related injuries had the lowest RR (0.30; 95% CI 0.17-0.52) when comparing pre- and post-intervention time periods. PMID- 21632035 TI - Amyotrophic lateral sclerosis: a neurovascular disease. AB - Amyotrophic lateral sclerosis (ALS) is a severe neurodegenerative disease with a complicated pathogenesis. Compelling evidence indicates impairment of all neurovascular unit components including the blood-brain and blood-spinal cord barriers (BBB/BSCB) in both patients and animal models, leading to classification of ALS as a neurovascular disease. The present review provides an updated analysis of the normal and impaired BBB/BSCB, focusing on the ALS-altered barrier. Here we describe the roles of cellular components, tight junctions, transport systems, cell interactions, cytokines, matrix metalloproteinases, and free radicals in the BBB/BSCB disruption, along with recent evidence from experimental and clinical ALS studies. The BBB/BSCB is a promising research area in ALS and this review will reveal some aspects of microvascular pathology in ALS and hopefully provide ideas for the development of new therapeutic strategies. PMID- 21632036 TI - Organoiridium complexes: efficient catalysts for the formation of sugar acetals and ketals. AB - [Cp(*)IrCl(2)](2) is used as an efficient promoter for the synthesis of sugar acetals and ketals with good to excellent yields. The catalyst is found to be general for a wide range of sugars. PMID- 21632037 TI - Self-mutilative behaviors in male substance-dependent inpatients and relationship with anger and aggression: mediator effect of childhood trauma. AB - The aim of this study was to investigate the relationship of self-mutilation (SM) with anger and aggression in male substance-dependent inpatients. Also, we wanted to evaluate the mediator effect of childhood trauma on these relationships while controlling variables such as age, substance of dependence (alcohol/drug), and negative effect. Participants were consecutively admitted 200 male substance dependent inpatients. Patients were investigated with the Self-mutilative Behaviour Questionnaire, the Childhood Trauma Reports, the Buss-Perry's Aggression Questionnaire, the State-Trait Anger Expression Inventory, the Beck Depression Inventory, and the State-Trait Anxiety Inventory. Rate of being married, current age, and age onset of regular substance use were lower, whereas being unemployed and history of childhood trauma (HCT) were higher in group with SM (n = 124, or 62.0%). Higher mean scale scores were found in SM group. Predictors of SM were being younger, impaired anger control, and physical aggression in logistic regression model. Being younger and the outward expression of anger (anger-out) predicted SM in the subgroup of patients without HCT, whereas being younger, severity of anger, and the inward expression of anger (anger-in) predicted SM in the subgroup of patients with HCT. Thus, to reduce self-mutilative behavior among substance-dependent patients, clinicians must improve anger control, particularly in younger patients. Type of strategy for coping with anger, which must be worked on, may differ in different subgroup patients, that is, focusing anger toward self among those with HCT, whereas anger toward others among those without. PMID- 21632038 TI - The symptom and function dimensions of the Global Assessment of Functioning (GAF) scale. AB - OBJECTIVE: The objective was to investigate the validity and clinical impact of the symptom and function dimensions of the Global Assessment of Functioning (GAF) in the Diagnostic and Statistical Manual of Mental Disorders (DSM), Fourth Edition. Is there any need for revision with respect to the DSM, Fifth Edition? MATERIAL: The sample comprised 2695 patients consecutively admitted to 14 different treatment units participating in the Norwegian Network of Personality Focused Treatment Programs from 1998 to 2007. METHODS: Convergent and discriminant validity of the symptom and function dimensions of GAF was analyzed by their associations with demographic variables, diagnostic status, and other self-reported variables assessing symptom distress, interpersonal problems, work and social impairment, and quality of life. RESULTS: The validity of the separate GAF dimensions was confirmed by discriminant and concurrent associations to other relevant clinical measures. However, the traditional GAF measure based on the lower score of either symptom or function level was found to serve well as a global indicator of symptom distress and social dysfunction. A substantial difference between the symptom and function score of GAF was found in about 10% of the cases; and when differences were found, functional impairment was most often more severe. CONCLUSION: This study confirms the validity of the 2 GAF dimensions. However, substantial differences between these dimensions are rarely occurring. We therefore recommend that the GAF scale be prolonged in the DSM, Fifth Edition, roughly in the same shape as in the DSM, Fourth Edition. PMID- 21632039 TI - Brazilian Portuguese validation of Mood Disorder Questionnaire. AB - BACKGROUND: The Mood Disorder Questionnaire (MDQ) is a screening instrument for bipolar spectrum disorders already validated in many languages. METHODS: Patients from 2 psychiatric outpatient facilities were diagnosed with bipolar disorder (BD) type I and II and major depression according to the mood module of the Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (SCID), Axis I Disorders--Clinician Version. In addition, a control group of healthy subjects was selected. The diagnostic interviews were used as the gold standard against which to investigate the performance of the MDQ. The MDQ was administered to 153 subjects, distributed among 4 groups. We analyzed the test reliability and discriminative capacity of the MDQ for the detection of patients with BD. RESULTS: Based on the SCID, Axis I Disorders--Clinician Version, 52 subjects (33.3%) presented a bipolar spectrum disorder (type I, II, or not otherwise specified), 48 (32.4%) were diagnosed as having unipolar depressive disorder, whereas 54 (35.3%) were unaffected by any type of psychiatric disorder (had no psychiatric disorder according to SCID results). The sensitivity for bipolar disorder was 0.72 (bipolar I disorder, 0.81; bipolar II disorder, 0.58; and bipolar disorder not otherwise specified, 0.69), with specificity of 0.95. The Brazilian Portuguese MDQ demonstrated adequate internal consistency (Cronbach alpha = .87). LIMITATIONS: Recruiting patients attending tertiary services may inflate the performance of the MDQ. CONCLUSIONS: The performance of the Brazilian Portuguese MDQ is comparable with other language validations. In a psychiatric outpatient sample, the Brazilian Portuguese MDQ proves to be a feasible and reliable screening instrument. PMID- 21632040 TI - Exploratory and confirmatory factor analyses of the Multidimensional Scale of Perceived Social Support in patients with schizophrenia. AB - INTRODUCTION: Social support promotes community adaptation of patients with mental illness by facilitating coping and competence. The Multidimensional Scale of Perceived Social Support (MSPSS) is a widely used measure of perceived social support that investigates 3 domains of support, from significant other, family, and friends. This study was conducted to provide evidence of the reliability and construct validity of the MSPSS in an adult Asian population with schizophrenia. METHODS: The MSPSS was self-rated by patients with schizophrenia. Reliability was determined using Cronbach alpha for internal consistency, and preliminary construct was examined using exploratory factor analysis (using principal component with varimax rotation). Four a priori hypothesized models were tested via confirmatory factor analysis: first-order 2-factor (2 models), and first order and higher order 3-factor models, using 5 goodness-of-fit indices. RESULTS: Internal reliability was high (Cronbach alpha of .90, .90, and .91 for support from significant others, family and friends, respectively) for the MSPSS. Exploratory factor analysis on the 12-item measure indicated 3-factor solution that supported the 3 theoretical dimensions of the MSPSS. The indices of goodness of fit confirmed the first-order and higher-order 3-factor confirmatory models of MSPSS by providing the best fit in this sample. CONCLUSION: Our findings demonstrate high internal consistency and construct validity of MSPSS in outpatients with schizophrenia, making it appropriate for assessing the perceived social support in this population. PMID- 21632041 TI - Association of insight with sociodemographic and clinical factors, quality of life, and cognition in Chinese patients with schizophrenia. AB - PURPOSE: The aims of this study were to assess insight in Chinese patients with schizophrenia and to identify its relationship with sociodemographic and clinical factors, cognition, and quality of life (QOL). METHODS: A cohort of 139 patients with clinically stable schizophrenia was selected by consecutively screening patients who had been diagnosed with schizophrenia and who were attending the outpatient department of a university-affiliated psychiatric hospital in China. Participants' sociodemographic and clinical characteristics, including psychotic and depressive symptoms and insight, as well as their social functioning, QOL, and flexibility of cognition, were assessed with standardized rating instruments. RESULTS: Thirty-three patients (23.7%) had good insight into their illness. In univariate analyses, poor insight was associated with the positive, negative, and general symptom scores of the Positive and Negative Syndrome Scale (PANSS), and with higher scores on the physical and mental components of QOL. In multivariate analysis, poor insight was independently associated with a higher negative symptom score on the PANSS, a shorter length of illness, and with a high score on the physical components of QOL. CONCLUSIONS: Poor insight is common in patients with clinically stable schizophrenia. In this study, insight was not associated with basic sociodemographic characteristics or had it any associations with depression or the flexibility of cognitive processes. PMID- 21632042 TI - Affective lability in bipolar disorder and borderline personality disorder. AB - BACKGROUND: The boundaries between the affective instability in bipolar disorder and borderline personality disorder have not been clearly defined. Using self report measures, previous research has suggested that the affective lability of patients with bipolar disorder and borderline personality disorder may have different characteristics. METHODS: We assessed the mood states of 29 subjects meeting Revised Diagnostic Interview for Borderlines and Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) criteria for BPD and 25 subjects meeting DSM-IV criteria for bipolar II disorder or cyclothymia using the Affective Lability Scale (ALS), the Affect Intensity Measure (AIM), and a newly developed clinician-administered instrument, the Affective Lability Interview for Borderline Personality Disorder (ALI-BPD). The ALI-BPD measures frequency and intensity of shifts in 8 affective dimensions. Subjects in the borderline group could not meet criteria for bipolar disorder; subjects in the bipolar/cyclothymia group could not meet criteria for BPD. RESULTS: Patients in the bipolar group had significantly higher scores on the euthymia-elation subscale of the ALS; patients in the BPD group had significantly higher scores on the anxiety-depression subscale of the ALS. Patients with bipolar disorder had significantly higher total AIM scores and significantly higher score on the AIM positive emotion subscale. In terms of frequency, patients in the borderline group reported the following: (1) significantly less frequent affective shifts between euthymia-elation and depression-elation on the ALI-BPD and (2) significantly more frequent shifts between euthymia-anger, anxiety-depression, and depression-anxiety. In terms of intensity, borderline patients reported the following: (1) significantly less intense shifts between euthymia-elation and depression-elation on the ALI-BPD and (2) significantly more intense shifts between euthymia-anxiety, euthymia-anger, anxiety-depression, and depression anxiety. CONCLUSION: The affective lability of patients with borderline and bipolar II/cyclothymic can be differentiated with respect to frequency and intensity using both self-report and clinician-administered measures. PMID- 21632043 TI - A computer model for the simulation of fiber-cell interaction in the alveolar region of the respiratory tract. AB - A mathematical model is presented that describes the health-endangering interaction of fibrous particles deposited in the human alveoli with alveolar fluids, cells, and tissues. As suggested by the theoretical approach, short fibers (diameter: 0.5 MUm, length: 2-10 MUm) are preferably ingested by alveolar macrophages and removed from the alveolar surface 10-15 days after exposure. Long (diameter: 0.5 MUm, length: 10-50 MUm) biopersistent fibers are not effectively cleared from the alveoli due to the repeated process of frustrated phagocytosis. Long biosoluble fibers also undergo a frustrated phagocytosis, with processes of extensive lysis leading to their significant shortening. The decrease in length causes the initiation of those clearance mechanisms that are efficient for short fibers. PMID- 21632044 TI - Computing the stability of steady-state solutions of mathematical models of the electrical activity in the heart. AB - Instabilities in the electro-chemical resting state of the heart can generate ectopic waves that in turn can initiate arrhythmias. We derive methods for computing the resting state for mathematical models of the electro-chemical process underpinning a heartbeat, and we estimate the stability of the resting state by invoking the largest real part of the eigenvalues of a linearized model. The implementation of the methods is described and a number of numerical experiments illustrate the feasibility of the methods. In particular, we test the methods for problems where we can compare the solutions with analytical results, and problems where we have solutions computed by independent software. The software is also tested for a fairly realistic 3D model. PMID- 21632045 TI - Mathematical modeling and sensitivity analysis of the integrated TNFalpha mediated apoptotic pathway for identifying key regulators. AB - TNFalpha-mediated apoptosis is one of the complex and tightly regulated cellular processes as it involves the activation of both pro- and anti-apoptotic signaling pathways. Thus, it is important to elucidate the molecular players of this process and their dynamics in order to gain an in-depth understanding of the mechanisms underlying apoptosis. To this end, we proposed an integrated model of TNFalpha-mediated apoptosis pathway in Type I cells, formulated based on the principles of mass action kinetics. The model includes major apoptotic modules the extrinsic and intrinsic pathways, the NFkappaB survival signaling and various regulatory mechanisms. We performed simulations and sensitivity analyses to study the role of NFkappaB pathway in regulating apoptosis, and identified IAP as one of the more potent regulators of apoptosis. PMID- 21632046 TI - Neuroimaging findings in frontotemporal lobar degeneration spectrum of disorders. AB - Frontotemporal lobar degeneration (FTLD) is a clinically and pathologically heterogeneous spectrum of disorders. In the last few years, neuroimaging has contributed to the phenotypic characterisation of these patients. Complementary to the clinical and neuropsychological evaluations, structural magnetic resonance imaging (MRI) and functional techniques provide important pieces of information for the diagnosis of FTLD. They also appear to be useful in distinguishing FTLD from patients with Alzheimer's disease (AD). Preliminary studies in pathologically proven cases suggested that distinct patterns of tissue loss could assist in predicting in vivo the pathological subtype. Recent years have also witnessed impressive advances in the development of novel imaging approaches. Diffusion tensor MRI and functional MRI have improved our understanding of the pathophysiology of the disease, and this should lead to the identification of additional useful markers of disease progression. This reviews discusses comprehensively the state-of-the-art of neuroimaging in the study of FTLD spectrum of disorders, and attempts to envisage which will be new neuroimaging biomarkers that could serve as surrogate measures of the underlying pathology. This will be central in the design of treatment trials of experimental drugs, which are likely to emerge in the near future, to target the pathological processes associated with this condition. PMID- 21632047 TI - Neurolaw and consciousness detection. PMID- 21632048 TI - Picture-word interference and the response-exclusion hypothesis. PMID- 21632049 TI - Non-native marine invertebrates are more tolerant towards environmental stress than taxonomically related native species: results from a globally replicated study. AB - To predict the risk associated with future introductions, ecologists seek to identify traits that determine the invasiveness of species. Among numerous designated characteristics, tolerance towards environmental stress is one of the most favored. However, there is little empirical support for the assumption that non-native species generally cope better with temporarily unfavorable conditions than native species. To test this concept, we ran five pairwise comparisons between native and non-native marine invertebrates at temperate, subtropical, and tropical sites. We included (natives named first) six bivalves: Brachidontes exustus and Perna viridis, P. perna and Isognomon bicolor, Saccostrea glomerata and Crassostrea gigas, two ascidians: Diplosoma listerianum and Didemnum vexillum as well as two crustaceans: Gammarus zaddachi and G. tigrinus. We simulated acute fluctuations in salinity, oxygen concentration, and temperature, while we measured respiration and survival rates. Under stressful conditions, non-native species consistently showed less pronounced deviations from their normal respiratory performance than their native counterparts. We suggest that this indicates that they have a wider tolerance range. Furthermore, they also revealed higher survival rates under stress. Thus, stress tolerance seems to be a property of successful invaders and could therefore be a useful criterion for screening profiles and risk assessment protocols. PMID- 21632050 TI - Baseline cerebral oximetry values in elderly patients with hip fractures: a prospective observational study. AB - AIM: This study was conducted to evaluate baseline cerebral tissue regional oxygen saturation (rSO(2)) values and identify risk factors related to severe rSO(2) reductions in elderly patients with hip fractures. PATIENTS AND METHODS: This was a prospective observational single-centre study on patients undergoing scheduled or urgent operation for isolated hip fracture. The study was approved by the Institution Ethics Committee, and all patients signed informed consent before entering the study. Data were collected on factors potentially related to baseline cerebral rSO(2). Data were analysed with Student's t-test, Pearson's correlation or multiple regression analysis as appropriate. RESULTS: Sixty-nine patients, aged (mean+/-standard deviation (SD)) 74+/-13 years participated. Left baseline rSO(2) was 60.09+/-10.20 and right baseline rSO(2) was 58.64+/-9.92. Baseline rSO(2)<45 was observed in 10.1% of patients on the left and 8.7% on the right side. Correlation between left- and right-side baseline cerebral rSO(2) was highly significant (r=0.852, p<0.001). Baseline cerebral rSO(2) had a positive, highly significant correlation with preoperative haematocrit (r=0.50, p<0.001) and arterial haemoglobin oxygen saturation (SpO(2)) (r=0.587, p<0.001), but correlation was negative with the American Society of Anesthesiologists (ASA) physical status (r=-0.42, p<0.001) and age (r=-0.39, p=0.001). Linear regression showed that preoperative haematocrit accounts for 23% (R(2)=0.23) of baseline rSO(2) variability, whereas preoperative haematocrit and SpO(2) combined account for 43.7% of rSO(2) variability (R(2)=0.437). Combined preoperative haematocrit+SpO(2)+age accounted for 51.3% (R(2)=0.513) of observed rSO(2) variability. CONCLUSION: Low baseline cerebral rSO(2) values are common in elderly hip fracture patients, despite normal haemodynamic and arterial saturation values. Preoperative haematocrit, SpO(2) and age explain a significant portion of cerebral rSO(2) variability. More studies are needed to validate our findings and assess the potential benefit of interventions aimed at improving cerebral rSO(2) in elderly hip fracture patients. PMID- 21632051 TI - What does it mean to engage in mental health promotion: does one size fit all? PMID- 21632052 TI - Protease activity in the multi-layered intra-luminal thrombus of abdominal aortic aneurysms. AB - INTRODUCTION: Rupture of an abdominal aortic aneurysm (AAA) is the cause of death in approximately 2% of men above 65 years. Most AAAs contain an intra-luminal thrombus (ILT), which is a potential source of proteases capable of degrading the underlying aneurysm wall. The AAA wall covered by a thick ILT shows more signs of matrix degradation compared to the wall free from ILT. The purpose of the present study was to evaluate the presence of protease activity in the ILT. MATERIALS AND METHODS: ILT and peripheral blood from 32 patients undergoing elective surgery were collected. The ILT was divided into abluminal, luminal, and a middle layer in between. Collagenases, gelatinases, elastase, and their inhibitors were measured using ELISA in protein extracts from these layers. Immunohistochemistry was used for identification of cells. RESULTS: Neutrophil leukocytes and platelets were mostly detected in the luminal layer of the ILT. MMP-9 and neutrophil elastase were also abundant in this layer but with low activity. High concentrations of TIMP-1 and PAI-1 were detected in the abluminal layer, while alpha 1 antitrypsin was mostly found in the luminal layer of the ILT. CONCLUSIONS: In AAA thick ILTs with multiple layers contain substantial amounts of proteases, but their activity is limited to the luminal layer. Proteases in the abluminal layer are mostly inactive, probably due to excess amounts of inhibitors and are consequently unable to directly participate in the pathogenesis of AAA. PMID- 21632053 TI - Arginine bioavailability ratios are associated with cardiovascular mortality in patients referred to coronary angiography. AB - OBJECTIVES: Arginine is the only source for nitric oxide (NO) synthesis. The bioavailability of NO plays a pivotal role in endothelial function and consequently in cardiovascular disease. The aim of the current study is to investigate the association of arginine bioavailability ratios with markers of endothelial function and cardiovascular mortality in patients referred to coronary angiography. METHODS: We investigated 2236 patients recruited within the LUdwigshafen RIsk and Cardiovascular Health (LURIC) study that were followed up for a median of 7.7 years. Arginine, ornithine and citrulline were chromatographically determined after precolumn-derivatisation followed by postcolumn continuous reaction with ninhydrin. Global arginine bioavailability (GABR) was calculated by arginine divided by the sum of ornithine plus citrulline. RESULTS: We observed a significant rise in cardiovascular mortality with decreasing GABR and arginine to ornithine ratio quartiles. The adjusted Cox proportional HRs for GABR were 1.27 (0.88-1.83), 1.27 (0.89-1.80) and 1.75 (1.24 2.45) for the 3rd, the 2nd and the 1st quartile respectively in comparison to the 4th quartile. The HRs for the quartiles of the arginine to ornithine ratio were 1.83 (1.25-2.67), 2.17 (1.50-3.20) and 2.02 (1.39-2.92) respectively. Patients with type 2 diabetes mellitus had a significantly lower GABR than persons without diabetes (0.88 +/- 0.23 vs. 0.94 +/- 0.24, p<0.001). GABR was found to be inversely correlated with endothelial markers as VCAM-1 (r=-0.301, p<0.001) or ICAM-1 (r=-0.136, p<0.001). CONCLUSIONS: GABR and the arginine to ornithine ratio are associated with markers of endothelial dysfunction and increased risk of cardiovascular mortality. Further studies are warranted to elucidate the pathobiology and clinical relevance of the arginine bioavailability ratios in cardio-metabolic diseases. PMID- 21632054 TI - Wave propagation in protein microtubules modeled as orthotropic elastic shells including transverse shear deformations. AB - Wave propagation along the microtubules is one of the issues of major concern in various microtubule cellular functions. In this study, the general wave propagation behavior in protein microtubules is investigated based on a first order shear deformation shell theory for orthotropic materials, with particular emphasis on the role of strongly anisotropic elastic properties of microtubules. According to experimental observation, the first-order shear deformation theory is used for the modeling of microtubule walls. A general displacement representation is introduced and a type of coupled polynomial eigenvalue problem is developed. Numerical examples describe the effects of shear deformation and rotary inertia on wave velocities in orthotropic microtubules. Finally, the influences of the microtubule shear modulus, axial external force, effective thickness and material temperature dependency on wave velocities along the microtubule protofilaments, helical pathway and radial directions are elucidated. Most results presented in the present investigation have been absent from the literature for the wave propagation in microtubules. PMID- 21632055 TI - On a phenomenological model for active smooth muscle contraction. AB - This paper presents a three-dimensional phenomenological model for the description of smooth muscle activation. A strain energy function is proposed as sum of the strain energy stored in the passive tissue, consisting of elastin and collagen, and an active calcium-driven energy related to the chemical contraction of the smooth muscle cells. Further, the proposed model includes the dispersions of the orientations of smooth muscle cells and collagen. These dispersions, measured in experiments, can be directly inserted into the model. The approach is implemented into the framework of the finite element method. Consequently, beside a validation with experiments the modelling concept is used for a three dimensional numerical study. PMID- 21632056 TI - Stress and strain analysis of contractions during ramp distension in partially obstructed guinea pig jejunal segments. AB - Previous studies have demonstrated morphological and biomechanical remodeling in the intestine proximal to an obstruction. The present study aimed to obtain stress and strain thresholds to initiate contraction and the maximal contraction stress and strain in partially obstructed guinea pig jejunal segments. Partial obstruction and sham operations were surgically created in mid-jejunum of male guinea pigs. The animals survived 2, 4, 7 and 14 days. Animals not being operated on served as normal controls. The segments were used for no-load state, zero stress state and distension analyses. The segment was inflated to 10 cmH(2)O pressure in an organ bath containing 37 degrees C Krebs solution and the outer diameter change was monitored. The stress and strain at the contraction threshold and at maximum contraction were computed from the diameter, pressure and the zero stress state data. Young's modulus was determined at the contraction threshold. The muscle layer thickness in obstructed intestinal segments increased up to 300%. Compared with sham-obstructed and normal groups, the contraction stress threshold, the maximum contraction stress and the Young's modulus at the contraction threshold increased whereas the strain threshold and maximum contraction strain decreased after 7 days obstruction (P<0.05 and 0.01). In conclusion, in the partially obstructed intestinal segments, a larger distension force was needed to evoke contraction likely due to tissue remodeling. Higher contraction stresses were produced and the contraction deformation (strain) became smaller. PMID- 21632057 TI - Anisotropic mechanical properties of ovine femoral periosteum and the effects of cryopreservation. AB - The mechanical properties of periosteum are not well characterized. An understanding of these properties is critical to predict the environment of pluripotent and osteochondroprogenitor cells that reside within the periosteum and that have been shown recently to exhibit a remarkably rapid capacity to generate bone de novo. Furthermore, the effects of cryopreservation on periosteal mechanical properties are currently unknown. We hypothesized that the periosteum is pre-stressed in situ and that the periosteum exhibits anisotropic material properties, e.g. the elastic modulus of the periosteum depends significantly on the direction of loading. We measured the change in area, axial length, and circumferential length of anterior, posterior, medial, and lateral fresh periosteal samples removed from underlying bone (t=0-16 h) as well as the average strain in axially and circumferentially oriented anterior periosteal samples subjected to tensile strain (0.004 mm/s) until failure. The elastic modulus was calculated from the resulting stress-strain curves. Tensile testing was repeated with axially aligned samples that had been slowly cryopreserved for comparison to fresh samples. Periosteal samples from all aspects shrank 44-54%, 33-47%, and 9 19% in area, axial length, and circumferential length, respectively. At any given time, the periosteum shrank significantly more in the axial direction than the circumferential direction. Tensile testing showed that the periosteum is highly anisotropic. When loaded axially, a compliant toe region of the stress-strain curve (1.93+/-0.14 MPa) is followed by a stiffer region until failure (25.67+/ 6.87 MPa). When loaded circumferentially, no toe region is observable and the periosteum remained compliant until failure (4.41+/-1.21 MPa). Cryopreservation had no significant effect on the elastic modulus of the periosteum. As the periosteum serves as the bounding envelope of the femur, anisotropy in periosteal properties may play a key role in modulating bone growth, healing and adaptation, in health, disease, and trauma. PMID- 21632058 TI - Development of a new QuEChERS method based on dry ice for the determination of 168 pesticides in paprika using tandem mass spectrometry. AB - This study describes a new QuEChERS method referred to as the dry ice partitioning QuEChERS method. This current method can be differentiated from the other QuEChERS methods in the sense that it uses dry ice rather than salts or buffers to extract and partition pesticides in the first extraction step. The dry ice-partitioning QuEChERS method consists of extraction method A (for detection of the acetonitrile layer) and extraction method B (for detection of both acetonitrile and aqueous layers). The extraction efficiency was then compared with the citrate-buffering QuEChERS method by means of recovery. Recovery values of the tested 168 pesticides were above 76%, with relative standard deviations of less than 20%. Certain problematic pesticides, including benfuracarb, carbosulfan, dichlofluanid, probenazole, pymetrozine, tolylfluanid, TFNA, and TFNG evidenced acceptable recoveries via the dry ice-partitioning QuEChERS method compared to the less than 70% recoveries of the citrate-buffering QuEChERS method examined herein. The matrix effect of paprika on the method developed herein was not significant, and matrix-matched calibration was performed well, with an r(2)>=0.99. The dry ice-partitioning QuEChERS method is capable of detecting the aqueous layer as well as the acetonitrile layer; this interesting feature makes it worth in application as an alternative QuEChERS method for the multiresidue analysis of pesticides within a broad polarity range in various matrices. PMID- 21632059 TI - Models for liquid-liquid partition in the system dimethyl sulfoxide-organic solvent and their use for estimating descriptors for organic compounds. AB - Partition coefficients for varied compounds were determined for the organic solvent-dimethyl sulfoxide biphasic partition system where the organic solvent is n-heptane or isopentyl ether. These partition coefficient databases are analyzed using the solvation parameter model facilitating a quantitative comparison of the dimethyl sulfoxide-based partition systems with other totally organic partition systems. Dimethyl sulfoxide is a moderately cohesive solvent, reasonably dipolar/polarizable and strongly hydrogen-bond basic. Although generally considered to be non-hydrogen-bond acidic, analysis of the partition coefficient database strongly supports reclassification as a weak hydrogen-bond acid in agreement with recent literature. The system constants for the n-heptane-dimethyl sulfoxide biphasic system provide an explanation of the mechanism for the selective isolation of polycyclic aromatic compounds from mixtures containing low polarity hydrocarbons based on the capability of the polar interactions (dipolarity/polarizability and hydrogen-bonding) to overcome the opposing cohesive forces in dimethyl sulfoxide that are absent for the interactions with hydrocarbons of low polarity. In addition, dimethyl sulfoxide-organic solvent systems afford a complementary approach to other totally organic biphasic partition systems for descriptor measurements of compounds virtually insoluble in water. PMID- 21632060 TI - Ionic liquid based three-phase liquid-liquid-liquid solvent bar microextraction for the determination of phenols in seawater samples. AB - For the first time, an ionic liquid based three-phase liquid-liquid-liquid solvent bar microextraction (IL-LLL-SBME) was developed for the analysis of phenols in seawater samples. The ionic liquid, 1-butyl-3-methylimidazolium hexafluorophosphate ([BMIM][PF(6)]), was used as the intermediary solvent for LLL SBME, enhancing the extraction efficiency for polar analytes. In the procedure, the analytes were extracted from the aqueous sample into the ionic liquid intermediary and finally, back-extracted into an aqueous acceptor solution in the lumen of the hollow fiber. The porous polypropylene membrane acted as a filter to prevent potential interfering materials from being extracted, and no additional cleanup was required. After extraction, the acceptor solution could be directly injected into a high-performance liquid chromatographic system for analysis. Six phenols, 2-nitrophenol, 4-chlorophenol, 2,3-dichlorophenol, 2,4-dichlorophenol, 2,4,6-trichlorophenol and pentachlorophenol were selected here as model compounds for developing and evaluating the method. The most influential extraction parameters were evaluated, including the ionic liquid, the composition of donor solution and acceptor solution, the extraction time and the extraction temperature, the effect of ionic strength, and the agitation speed. Under the most favorable extraction parameters, the method showed good linearity (from 0.05 50 to 0.5-50 MUg/L, depending on the analytes) and repeatability of extractions (RSD below 8.3%, n=5). The proposed method was compared to conventional three phase LLL-SBME and ionic liquid supported hollow fiber protected three-phase liquid-liquid-liquid microextraction, and showed higher extraction efficiency. The proposed method was demonstrated to be a simple, fast, and efficient method for the analysis of phenols from environmental water samples. PMID- 21632061 TI - Simultaneous determination of trace sterols in complicated biological samples by gas chromatography-mass spectrometry coupled with extraction using beta sitosterol magnetic molecularly imprinted polymer beads. AB - In this paper, an efficient and sensitive analytical method for the simultaneous determination of three trace sterols including ergosterol, stigmasterol and beta sitosterol in complicated biological samples was developed by gas chromatography mass spectrometry (GC-MS) coupled with extraction using novel beta-sitosterol magnetic molecularly imprinted polymer (mag-MIP) beads. Physical tests suggested that beta-sitosterol mag-MIP beads prepared by a rapid microwave synthesis method possessed the porous morphology, narrow size distribution, stable chemical and thermal property. Due to the greatly enlarging surface area and the strong recognition to the target molecules, beta-sitosterol mag-MIP beads have a higher enrichment factor for beta-sitosterol (~20-fold) and the higher selectivity for beta-sitosterol and its analogs than that of beta-sitosterol magnetic nonimprinted polymer (mag-NIP) beads. Under the optimum analytical conditions, all the target compounds achieved good chromatographic separation and sensitive detection without matrix interference. It was interesting that three target sterols were actually found in mushroom samples, and stigmasterol and beta sitosterol were actually found in serum and watermelon samples. The recoveries of spiked sample tests were in range of 71.6-88.2% with RSDs of 2.4-10.0% (n=3). This method is reliable and applicable for the simultaneous determination of trace sterols in real biological samples based on the beta-sitosterol mag-MIP bead extraction. PMID- 21632062 TI - Characterization of carboxylic acids in atmospheric aerosols using hydrophilic interaction liquid chromatography tandem mass spectrometry. AB - A sensitive hydrophilic interaction liquid chromatography/electrospray ionization mass spectrometry (HILIC/ESI-MS/MS) method was developed for determination of selected aliphatic (i.e. malonic, succinic, glutaric, adipic, pimelic, suberic, azelaic, maleic, fumaric, glycolic and pyruvic acid), alicyclic (i.e. cis-pinonic and pinic acid) and aromatic (i.e. trimesic, phthalic acid and its isomers) carboxylic acids. Analytes were separated on an amide column using a gradient elution with a 10mM constant ionic strength mobile phase containing acetonitrile and aqueous ammonium acetate buffer (pH 5.0). The influence of the buffer type, pH, polar modifier and temperature on analyte retention under HILIC was studied. Static sonication-assisted solvent extraction was optimized for sample preparation prior to analysis. The recoveries obtained were higher than 90% for most analytes. The method was proven to be sensitive with limits of detection ranged from 0.03 to 16.0 MUg/L in selected reaction monitoring mode (SRM). The repeatability and intermediate precision of the method, expressed as RSD (%) of the peak area ratio between analytes and their internal standards were generally lower than 5%. The method was successfully applied for determination of the studied acids in samples of ambient aerosol particles. A big advantage of the new method is also its ability to detect and separate the isobaric compounds of the selected carboxylic acids. Our results demonstrate that the method is specific and sensitive for the determination of a wider range of polar carboxylic acids at low concentrations in complex samples of aerosol particles. PMID- 21632063 TI - Narrative skills in Swedish children with language impairment. AB - This study investigated the development of narrative skills in Swedish children with language impairment between age 5 and age 10. Seventeen children with LI and two control groups of age peers with typical development participated in a picture elicited story telling task. Analyses included measures of story content, cohesion and grammar. Our subjects showed development in different areas from age 5 to 10, but they did not perform at the level of the controls at age 10 on number of different verbs used and percent grammatically correct C-units. We conclude that preschool children with LI develop in their narrative skills over time, but not to the level of their age-peers at age 10. LEARNING OUTCOMES: The reader will be able to describe areas of vulnerability in Swedish-speaking children with language impairment in general, and related to narration in particular. Furthermore, the reader will be able to describe similarities in narrative skills between Swedish-speaking and English-speaking children with language impairment. PMID- 21632064 TI - The treatment of minor depression with St. John's Wort or citalopram: failure to show benefit over placebo. AB - This paper presents new data addressing two important controversies in psychiatry: the construct of Minor Depression (MinD) and the efficacy of St. John's Wort for milder forms of depressive disorders. Data are from a three-arm, 12 week, randomized clinical trial of investigating the efficacy of St. John's Wort (810 mg/day), citalopram (20 mg/day), or placebo for acute treatment of MinD. Due to a high placebo response on all outcome measures, neither St. John's Wort nor citalopram separated from placebo on change in depressive symptom severity, quality of life, or well-being. However, systematic assessment of potential adverse effects (AEs) led to three important observations: (1) prior to the administration of study compound, 60% of subjects endorsed items that would be characterized as AEs once study compound was administered, (2) St. John's Wort and citalopram were each associated with a significant number of new or worsening AEs during treatment, and (3) using a structured interview for identifying AEs at baseline and during treatment is informative. MinD was not responsive to either a conventional antidepressant or a nutraceutical, and both compounds were associated with a notable side effects burden. Other treatment approaches for MinD should be investigated. PMID- 21632065 TI - Correlation of cerebral spinal fluid pH and HCO3- with disease progression in ALS. AB - Amyotrophic lateral sclerosis (ALS) is a progressive and fatal disease, which is characterized by progressive degeneration of spinal and bulbar innervating motor neurons. However, the underlying mechanisms of motor neuron death remain poorly understood. Several candidate disease biomarkers have been detected in cerebrospinal fluid of ALS patients. The present study analyzed various cerebral spinal fluid gas parameters in ALS patients and compared these values to controls, as well as patients with cervical spondylosis, Parkinson syndrome, and spinocerebellar degeneration. Cerebral spinal fluid pH positively correlated with the ALS functional rating scale in total and limb-type ALS patients. In addition, cerebral spinal fluid pH positively correlated with shorter disease duration (less than 22 weeks). These results suggested that cerebral spinal fluid pH provides a biomarker for ALS and could reflect mechanisms of disease progression in ALS patients. PMID- 21632066 TI - Proximate environmental drivers of coral communities at Palmyra Atoll: establishing baselines prior to removing a WWII military causeway. AB - A management proposal aims to partly remove a WWII military causeway at Palmyra Atoll to improve lagoon water circulation and alleviate sedimentation stress on the southeast backreef, an area of high coral cover and diversity. This action could result in a shift in sedimentation across reef sites. To provide management advice, we quantified the proximate environmental factors driving scleractinian coral cover and community patterns at Palmyra. The proportion of fine sedimentation was the optimal predictor of coral cover and changes in community structure, explaining 23.7% and 24.7% of the variation between sites, respectively. Scleractinian coral cover was negatively correlated with increases in fine sedimentation. Removing the causeway could negatively affect the Montipora corals that dominate the western reef terrace, as this genus was negatively correlated with levels of fine sedimentation. The tolerance limits of corals, and sediment re-distribution patterns, should be determined prior to complete removal of the causeway. PMID- 21632067 TI - [Relationship between odontogenic infections and infective endocarditis]. AB - Revised guidelines for the prevention of infective endocarditis published by national and international associations in the last years do not support the indiscriminate use of antibiotic prophylaxis for dental procedures. However, some of them still recommend its use in high-risk patients before dental treatments likely to cause bleeding. Given the high prevalence of bacteremia of dental origin due to tooth-brushing, mastication or other daily activities, it appears unlikely that infective endocarditis from oral microorganisms can be completely prevented. A good oral health status and satisfactory level of oral hygiene are sufficient to control the consequences of the systemic spread of oral microorganisms in healthy individuals. However, caution is still needed and prophylactic antibiotics must be administered to susceptible or medically compromised patients. This review briefly outlines the current concepts of odontogenic bacteremia and antibiotic prophylaxis for patients undergoing dental treatment. PMID- 21632068 TI - [Laminopathies. Nuclear lamina diseases]. AB - Laminopathies are a group of diseases that share wrong codification of lamins, building proteins of the nuclear lamina. Different tissues are affected in those disorders: striated muscle, adipose tissue, central or peripheral nervous system and aging process. Emery-Dreifuss muscular dystrophy and Hutchinson-Gildford Progery Syndrome are two examples of laminopathies. Other diseases, due to mutations in different genes, impair lamins function by a direct or an indirect way and they are frequently considered together. The last decade has seen an increasing interest and scientific advances on laminopathies that will allow us to answer key questions regarding metabolism, insulin resistance, sudden death and aging. Laminopathies are reviewed in this article from a molecular, pathogenic and clinical point of view. PMID- 21632069 TI - Effects of leptin and adiponectin on pancreatic beta-cell function. AB - Leptin and adiponectin are hormones secreted from adipocytes that have important roles in metabolism and energy homeostasis. This review evaluates the effects of leptin and adiponectin on beta-cell function by analyzing and compiling results from human clinical trials and epidemiologic studies as well as in vitro and in vivo experiments. Leptin has been shown to inhibit ectopic fat accumulation and thereby prevent beta-cell dysfunction and protect the beta-cell from cytokine- and fatty acid-induced apoptosis. However, leptin suppresses insulin gene expression and secretion as well as glucose transport into the beta-cell. Adiponectin stimulates insulin secretion by enhancing exocytosis of insulin granules and upregulating the expression of the insulin gene; however, this effect depends on the prevailing glucose concentration and status of insulin resistance. In addition, adiponectin has antiapoptotic properties in beta-cells. Available evidence concerning the role of these adipokines on insulin secretion, insulin gene expression, and apoptosis is not always entirely consistent; and many fundamental questions remain to be answered by future studies. PMID- 21632070 TI - Comparison of two methods using plasma triglyceride concentration as a surrogate estimate of insulin action in nondiabetic subjects: triglycerides * glucose versus triglyceride/high-density lipoprotein cholesterol. AB - The objective was to compare relationships between insulin-mediated glucose uptake and surrogate estimates of insulin action, particularly those using fasting triglyceride (TG) and high-density lipoprotein cholesterol (HDL-C) concentrations. Insulin-mediated glucose uptake was quantified by determining the steady-state plasma glucose (SSPG) concentration during the insulin suppression test in 455 nondiabetic subjects. Fasting TG, HDL-C, glucose, and insulin concentrations were measured; and calculations were made of the following: (1) plasma concentration ratio of TG/HDL-C, (2) TG * fasting glucose (TyG index), (3) homeostasis model assessment of insulin resistance, and (4) insulin area under the curve (insulin-AUC) during a glucose tolerance test. Insulin-AUC correlated most closely with SSPG (r ~ 0.75, P < .001), with lesser but comparable correlations between SSPG and TG/HDL-C ratio, TyG index, homeostasis model assessment of insulin resistance, and fasting TG and insulin (r ~ 0.60, P < .001). Calculations of TG/HDL-C ratio and TyG index correlated with SSPG concentration to a similar degree, and the relationships were comparable to estimates using fasting insulin. The strongest relationship was between SSPG and insulin-AUC. PMID- 21632071 TI - Deletion of interleukin-6 improves pyruvate tolerance without altering hepatic insulin signaling in the leptin receptor-deficient mouse. AB - Obesity is associated with increased circulating interleukin-6 (IL-6), which may contribute to hepatic insulin resistance by impairing insulin receptor signaling. This study was designed to assess the impact of the systemic absence of IL-6 on the development of insulin resistance and glucose intolerance in an obese mouse model. Systemic insulin, glucose, and pyruvate tolerance tests were performed in IL-6 knockout (IL6KO) mice that had been crossed with a genetically obese (leptin receptor-deficient mouse model [Lep(db)]) mouse model. Real-time reverse transcriptase polymerase chain reaction and Western blot analysis assessed cellular and molecular markers of insulin signaling, inflammation, and metabolism. Absence of IL-6 did not improve systemic glucose or insulin tolerance, but Lep(db) * IL6KO mice displayed a smaller blood glucose increase following a pyruvate challenge. These results suggest that loss of IL-6 in the context of obesity may locally reduce hepatic glucose production from a gluconeogenic precursor. Hepatic insulin-dependent insulin receptor autophosphorylation, Akt activation, and FoxO1 phosphorylation were similar between Lep(db) * IL6KO mice and Lep(db) controls. Basal gene expression of the gluconeogenic enzyme phosphoenolpyruvate carboxykinase was reduced in male Lep(db) * IL6KO mice relative to Lep(db) controls; but gene expression of another regulatory enzyme, glucose-6-phosphatase, remained unaltered. Absence of IL-6 reduced gene expression of serum amyloid A and RelA in female Lep(db) mice, but did not alter hepatic triglyceride accumulation or lipogenic gene expression. Overall, our results suggest that IL-6 may be detrimental in obesity by contributing to elevated hepatic glucose output. PMID- 21632072 TI - Impaired adrenergic- and corticotropic-axis outflow during exercise in chronic obstructive pulmonary disease. AB - Exercise stimulates coordinated release of the sympathoadrenal hormones adrenocorticotropic hormone (ACTH), cortisol, norepinephrine (NE), and epinephrine (Epi). The study hypothesis was that chronic obstructive pulmonary disease (COPD) is marked by heightened sympathoadrenal outflow at comparable relative workloads. The location of the study was at a clinical research unit. Eight healthy men and 9 men with stable COPD (forced expiratory volume at 1 second <75% predicted) were studied. Volunteers rested (baseline) or exercised at individual submaximal (35% +/- 5%) or maximal oxygen consumption. Blood was sampled every 2 minutes for 40 minutes concurrently. Two-way analysis of covariance was applied to examine group (healthy/COPD) and exercise (3 levels) effects on ACTH, cortisol, NE, and Epi release and regularity (estimable by approximate entropy). The timing of peak hormone concentrations was Epi, 14 minutes; NE, 16 minutes; ACTH, 22 minutes; and cortisol, 34 minutes in both cohorts. Type of exercise regimen influenced all 4 hormones (each P < .001), and subject group (control vs COPD) affected cortisol (P < .001) and Epi (P = .048) responses. Exercise regimen and group together controlled ACTH, cortisol, and Epi (each P < .001), but not NE, responses. In particular, endocrine responses were attenuated in COPD compared with control subjects. Approximate entropy analysis also identified loss of maximal exercise-induced ACTH-secretory regularity in COPD patients (P = .042). These outcomes demonstrate impaired rather than augmented exercise-associated sympathocorticotropic-axis outflow in patients with COPD even when outcomes are normalized to maximal oxygen consumption, suggesting that factors other than fitness are at work. PMID- 21632073 TI - Genetic deficiency of apolipoprotein D in the mouse is associated with nonfasting hypertriglyceridemia and hyperinsulinemia. AB - Apolipoprotein D (ApoD) is an atypical apolipoprotein with an incompletely understood function in the regulation of triglyceride and glucose metabolism. We have demonstrated that elevated ApoD production in mice results in improved postprandial triglyceride clearance. This work studies the role of ApoD deficiency in the regulation of triglyceride and glucose metabolism and its dependence on aging. We used ApoD knockout (ApoD-KO) mice of 3 and 21 months of age. Body weight and food intake were measured. Hepatic histology, triglyceride content, lipoprotein lipase levels, and plasma metabolites were studied. Phenotypic characterization of glucose metabolism was performed using glucose tolerance test. beta-Cell mass, islet volume, and islet number were analyzed by histomorphometry. Apolipoprotein D deficiency results in nonfasting hypertriglyceridemia in young (P = .01) and aged mice (P = .002). In young ApoD KO mice, hypertriglyceridemia was associated with 30% to 50% increased food intake in nonfasting and fasting conditions, respectively, without changes in body weight. In addition, lipoprotein lipase levels were reduced by 35% in adipose tissue (P = .006). In aged ApoD-KO mice, hypertriglyceridemia was not associated with changes in food intake or body weight, whereas hepatic triglyceride levels were reduced by 35% (P = .02). Furthermore, nonfasting plasma insulin levels were elevated by 2-fold in young (P = .016) and aged (P = .004) ApoD-KO mice, without changes in blood glucose levels, glucose tolerance, beta cell mass, or islet number. These findings underscore the importance of ApoD in the regulation of plasma insulin levels and triglyceride metabolism, suggesting that ApoD plays an important role in the pathogenesis of dyslipidemia. PMID- 21632074 TI - Serum adiponectin levels and tissue expression of adiponectin receptors are associated with risk, stage, and grade of colorectal cancer. AB - Adiponectin has been associated with colorectal cancer (CRC) risk. This study aims to investigate the association of both adiponectin and tissue expression of its receptors with CRC risk as well as clinicopathological characteristics, notably stage and grade. Determination of serum adiponectin and immunohistochemical expression of adiponectin receptors in adenocarcinoma/normal colorectal tissue was performed in samples from 104 newly diagnosed CRC patients and 208 age- and sex-matched controls. Multiple logistic regression odds ratios and 95% confidence intervals for CRC risk were derived, controlling for a series of covariates. Serum adiponectin was negatively associated with CRC risk (odds ratio, 0.72; confidence interval, 0.53-0.99) and also with tumor grade (P = .05). Expression of both adiponectin receptors was stronger in adenocarcinoma vs normal tissue (P = .001). AdipoR1 expression was negatively associated with nodal stage (P = .03); AdipoR2 expression was positively associated with tumor, node, metastasis stage (P = .01). Established positive associations with red meat consumption and diabetes, and negative associations with physical exercise and plant food consumption were confirmed along with a more than 60% higher risk associated with central obesity. Adiponectin levels and tissue expression of hormonal receptors seem to be associated not only with CRC risk but also with components of clinicopathological characteristics; given power limitations, these results should be interpreted with caution. The exact nature of the association and the underlying pathophysiological mechanisms need to be further examined in large prospective studies assessing adiponectin and its receptors as novel targets for exploring CRC growth. PMID- 21632075 TI - Resveratrol attenuates high-fat diet-induced insulin resistance by influencing skeletal muscle lipid transport and subsarcolemmal mitochondrial beta-oxidation. AB - Although resveratrol (RES) is implicated in the regulation of insulin sensitivity in rodents, the exact mechanism underlying this effect remains unclear. Therefore, we sought to investigate how RES affects skeletal muscle lipid transportation and lipid oxidation of subsarcolemmal (SS) and intermyofibrillar (IMF) mitochondrial populations in high-fat diet (HFD)-induced insulin resistance (IR) rats. Systemic and skeletal muscle insulin sensitivity together with expressions of several genes related to mitochondrial biogenesis and skeletal muscle lipid transportation was studied in rats fed a normal diet, an HFD, and an HFD with intervention of RES for 8 weeks. Citrate synthase (CS), electron transport chain (ETC) activities, and several enzymes for mitochondrial beta oxidation were assessed in SS and IMF mitochondria from tibialis anterior muscle. The HFD-fed rats exhibited obvious systemic and skeletal muscle IR as well as intramuscular lipid accumulation. SIRT1 activity and expression of genes related to mitochondrial biogenesis were greatly declined, whereas the gene for lipid transportation, FAT/CD36, was upregulated (P < .05). Subsarcolemmal but not IMF mitochondria displayed lower CS, ETC, and beta-oxidation activities. By contrast, RES treatment protected rats against diet-induced intramuscular lipid accumulation and IR, increased SIRT1 activity and mitochondrial biogenesis, and reverted the decline in SS mitochondrial CS and ETC activities. Importantly, although expression of FAT/CD36 was increased (11%, P < .05), activities of SS mitochondrial beta-oxidation enzymes were largely enhanced (41%~67%, P < .05). This study suggests that RES ameliorates insulin sensitivity consistent with an improved balance between skeletal muscle lipid transportation and SS mitochondrial beta-oxidation in HFD rats. PMID- 21632076 TI - Effects of nonenzymatic glycation and fatty acids on functional properties of human albumin. AB - Human serum albumin nonenzymatically condenses with glucose to form stable Amadori adducts that are increased with the hyperglycemia of diabetes. The present study evaluated the influence of fatty acids, which are major endogenous ligands, on albumin glycation and of glycation on albumin conformation and exogenous ligand binding. Physiologic concentrations of palmitate, oleate, and linoleate reduced the ability of albumin to form glucose adducts, whereas glycation decreased intrinsic fluorescence, lowered the affinity for dansylsarcosine, and diminished the fatty acid-induced increase in limiting fluorescence of protein-bound warfarin that was observed with nonglycated albumin. The findings indicate that fatty acids impede the ability of albumin to undergo Amadori glucose modification and induce conformational changes affecting exogenous ligand binding, and that nonenzymatic glycation of albumin induces alterations in structural and functional properties that may have import in lipid transport and atherogenesis. PMID- 21632077 TI - Relationship between glucose volume of distribution and the extracellular space: a multiple tracer study. AB - Although the use of radioisotopes in the investigation of glucose metabolism dates back more than 50 years, several relevant quantitative aspects have not been definitively determined. These include the volume of distribution (V(d)) of glucose and recycling of glucose radioisotopes from liver glycogen. These problems are further complicated by methodological issues such as the following: (1) glucose tracers have different metabolic fates that may influence volume estimates, and (2) the calculation method needs to be based on physical principles to avoid some limitations of compartmental models. To address these issues, we administered boluses of an extracellular marker ([1-(14)C]-l-glucose, 30 MUCi) and 2 glucose tracers ([2-(3)H]-d-glucose and [3-(3)H]-d-glucose, 120 MUCi of each), followed by a 1-mg glucagon bolus (in the presence of somatostatin) 245 minutes later, in conscious beagles to account for potential problems in recycling of the label through glycogen. We used modeling methods based on physical principles (circulatory model), which yield volume estimates with a clear physiological interpretation. Glucose V(d) (mL/kg) were 204 ([1 (14)C]-l-glucose), 191 ([2-(3)H]-d-glucose), and 206 ([3-(3)H]-d-glucose). These values were not different and correlated. The amount of recycled [3-(3)H]-d glucose in response to glucagon was small (~1.7% of the injected tracer dose). An additional result of this analysis is the determination of the parameters of the circulatory model in beagles for the standard [3-(3)H]-d-glucose tracer. Using multiple tracers in beagles and calculation methods based on physical principles, we have provided direct proof that the glucose V(d) equals the extracellular space in beagles under basal conditions. PMID- 21632078 TI - Portal vein and systemic adiponectin concentrations are closely linked with hepatic glucose and lipoprotein kinetics in extremely obese subjects. AB - Low systemic plasma adiponectin concentrations are associated with abnormalities in hepatic glucose and lipoprotein metabolism in obese people. However, the relationship between the delivery of adiponectin to the liver via the portal vein and hepatic glucose and lipoprotein metabolism is not known. We examined the relationship between hepatic substrate metabolism (glucose rate of appearance into plasma and hepatic very low-density lipoprotein [VLDL]-triglyceride [TG] and VLDL-apolipoprotein B-100 [apoB-100] secretion rates, determined by using stable isotope-labeled tracer techniques) and portal vein adiponectin concentration, in 8 insulin-resistant, extremely obese subjects (body mass index, 65 +/- 7 kg/m(2)). Portal vein adiponectin concentration was inversely associated with basal glucose rate of appearance (r = -0.820, P = .013) and VLDL-TG (r = -0.823, P = .012) and VLDL-apoB-100 (r = -0.787, P = .020) secretion rates. Very similar correlations were obtained for radial artery adiponectin as a result of a mirroring relationship between portal and arterial adiponectin concentrations (r = 0.899, P = .002) and the absence of significant arteriovenous concentration differences (P = .570). Insulin resistance, assessed with the homeostasis model assessment score, was also strongly associated with hepatic glucose and lipid metabolic parameters, as well as with adiponectin concentrations in the portal vein and radial artery. These results suggest that adiponectin delivery to the liver, whether via the portal or the systemic circulation, may be an important regulator of basal hepatic glucose, VLDL-TG, and VLDL-apoB-100 production rates in obese people, possibly through direct effects on the liver or changes in hepatic insulin sensitivity. However, portal vein adiponectin does not appear to be superior to arterial adiponectin as a marker of hepatic metabolic dysregulation. Additional studies are needed to elucidate the mechanism(s) responsible for the strong association we observed between adiponectin and hepatic substrate metabolism. PMID- 21632079 TI - Bone remodeling and calcium homeostasis in patients with spinal cord injury: a review. AB - Patients with spinal cord injury exhibit early and acute bone loss with the major functional consequence being a high incidence of pathological fractures. The bone status of these patients is generally investigated by dual-energy x-ray absorptiometry, but this technique does not reveal the pathophysiological mechanism underlying the bone loss. Bone cell activity can be indirectly evaluated by noninvasive techniques, including measurement of specific biochemical markers of bone formation (such as osteocalcin or bone-alkaline phosphatase) and resorption (such as procollagen type I N- or C-terminal propeptide). The bone loss in spinal cord injury is clearly due to an uncoupling of bone remodeling in favor of bone resorption, which starts just after the injury and peaks at about 1 to 4 months. Beyond 6 months, bone resorption activity decreases progressively but remains elevated for many years after injury. Conversely, bone formation is less affected. Antiresorptive treatment induces an early and acute reduction in bone resorption markers. Level of injury and health-related complications do not seem to be implicated in the intensity of bone resorption. During the acute phase, the hypercalcemic status is associated with the suppression of parathyroid hormone and vitamin D metabolites. The high sensitivity of these markers after treatment suggests that they can be used for monitoring treatment efficacy and patient compliance. The concomitant use of bone markers and dual-energy x-ray absorptiometry may improve the physician's ability to detect patients at risk of severe bone loss and subsequent fractures. PMID- 21632080 TI - Serum adiponectin and leptin in relation to risk for preeclampsia: results from a large case-control study. AB - Conditions resulting in insulin resistance, as well as metabolic, immune, and angiogenic perturbations, have been associated with an increased risk of preeclampsia (PE). Our purpose was to assess whether the adipose tissue-secreted hormones adiponectin, which has immune-modulating, metabolic, and angiogenic properties, and leptin, which reflects overall fat mass, are associated with PE risk. We performed a case-control design study within a hospital-based cohort of 368 pregnant women (106 with PE and 262 controls; mean age, 26.6 +/- 6.8 years; mean gestational age at admission, 38.2 +/- 2.8 weeks) between March 2005 and August 2007 at the Hospital of Pennsylvania University. Serum adiponectin and leptin were measured by radioimmunoassay. Statistical analysis of data was performed using simple and multiple regression analyses. No significant differences in adiponectin or leptin levels between preeclamptic and control pregnant women emerged in univariate analyses (P = .57 and P = .15, respectively). Among preeclamptic women, there were also no differences in adipokines between those with mild and severe disease. Serum adiponectin and leptin were not associated with higher risk of PE before and after adjustment for maternal age, race, primigravida, smoking status, body mass index at screening, gestational age at admission, history of PE, chronic hypertension, and gestational diabetes (odds ratio, 0.93; 95% confidence interval, 0.83-1.04 and odds ratio, 1; 95% confidence interval, 0.97-1.03, respectively). Maternal serum adiponectin and leptin levels, drawn at the time of PE diagnosis, were not associated with PE. PMID- 21632081 TI - Functional effects of DAX-1 mutations identified in patients with X-linked adrenal hypoplasia congenita. AB - X-linked adrenal hypoplasia congenita with hypogonadotropic hypogonadism and adrenal insufficiency is a rare disorder caused by mutations of DAX-1. In this study, we investigated the functional defects of DAX-1 caused by mutations identified in 3 unrelated Korean patients with adrenal hypoplasia congenita. The DAX-1 gene was directly sequenced using genomic DNA isolated from peripheral blood leukocytes. The functional defects of DAX-1 caused by mutations were evaluated using an in vitro promoter assay. After mutagenesis of DAX-1 complementary DNA in the pcDNA3.1 vector, steroidogenic factor 1 and the promoter region of steroidogenic acute regulatory protein (StAR) genes in pGL4.10[luc2] were transiently cotransfected into human embryonic kidney 293 cells, followed by luminometry measurements of the luciferase activity of StAR. Mutation analysis of 3 patients revealed p.L386delfsX2, p.W105X, and p.Q252X mutations of the DAX-1 gene. The mutant DAX-1 proteins showed lower repressive activity on the StAR gene promoter when compared with normal DAX-1. Nonsense and frameshift mutations of the DAX-1 gene partially eliminated the ability of DAX-1 to repress the transcription of StAR in an in vitro assay. PMID- 21632082 TI - Antifungal metabolites from fungal endophytes of Pinus strobus. AB - The extracts of five foliar fungal endophytes isolated from Pinus strobus (eastern white pine) that showed antifungal activity in disc diffusion assays were selected for further study. From these strains, the aliphatic polyketide compound 1 and three related sesquiterpenes 2-4 were isolated and characterized. Compound 2 is reported for the first time as a natural product and the E/Z conformational isomers 3 and 4 were hitherto unknown. Additionally, the three known macrolides; pyrenophorol (5), dihydropyrenophorin (6), and pyrenophorin (7) were isolated and identified. Their structures were elucidated by spectroscopic analyses including 2D NMR, HRMS and by comparison to literature data where available. The isolated compounds 1, 2, and 5 were antifungal against both the rust Microbotryum violaceum and Saccharomyces cerevisae. PMID- 21632083 TI - Distribution of lignin and its coniferyl alcohol and coniferyl aldehyde groups in Picea abies and Pinus sylvestris as observed by Raman imaging. AB - Wood cell wall consists of several structural components, such as cellulose, hemicelluloses and lignin, whose concentrations vary throughout the cell wall. It is a composite where semicrystalline cellulose fibrils, acting as reinforcement, are bound together by amorphous hemicelluloses and lignin matrix. Understanding the distribution of these components and their functions within the cell wall can provide useful information on the biosynthesis of trees. Raman imaging enables us to study chemistry of cell wall without altering the structure by staining the sample or fractionating it. Raman imaging has been used to analyze distributions of lignin and cellulose, as well as the functional groups of lignin in wood. In our study, we observed the distribution of cellulose and lignin, as well as the amount of coniferyl alcohol and aldehyde groups compared to the total amount of lignin in pine (Pinus sylvestris) and spruce (Picea abies) wood samples. No significant differences could be seen in lignin and cellulose distribution between these samples, while clear distinction was observed in the distribution of coniferyl alcohols and coniferyl aldehyde in them. These results could provide valuable insight on how two similar wood species control biosynthesis of lignin differently during the differentiation of cell wall. PMID- 21632084 TI - Brucellosis in camels. AB - Camels are highly susceptible to brucellosis caused by Brucella melitensis and Brucella abortus. Difficulties can arise in diagnosis of camel brucellosis, especially as this disease provokes only few clinical signs in contrast to its clinical course in cattle. Because none of the commonly used serological test can be perceived as a perfect test for Brucella diagnosis in camel and most serological tests used for camels have been directly transposed from cattle without adequate validation, an incorrect diagnosis may occur when diagnosis is based on serology alone. Of imminent concern is the fact that brucellosis can be easily transmitted from animals or their products to humans mainly via milk. In many developing countries in the arid areas of Asia and Africa, camels are still the most important productive livestock for nomadic populations. Therefore, we reviewed the literatures on camel brucellosis to highlight the epidemiologic, economic and public health impact of camel brucellosis as a basis for designing effective control strategies. PMID- 21632085 TI - Severe canine influenza in dogs correlates with hyperchemokinemia and high viral load. AB - Canine influenza virus (CIV) is an emerging pathogen that causes acute respiratory disease in dogs. To better understand the mechanism(s) responsible for the virulence of the virus, we conducted immunological, virological, clinical, and histopathological analyses in CIV-infected dogs. CIV replicated efficiently in the respiratory system of dogs and caused severe respiratory disease. Notably, the infection induced the marked elevation and sustained expression of chemokines that resulted in severe bronchointerstitial pneumonia with extensive neutrophil infiltration. In clinicopathological findings, CIV infection resulted in regenerative anemia, perhaps due to pulmonary hemorrhage. The observations indicate that active replication of CIV in the canine respiratory system results in intense inflammatory responses central to the pathogenesis of H3N2 CIV. PMID- 21632086 TI - Heavy metal removal in anaerobic semi-continuous stirred tank reactors by a consortium of sulfate-reducing bacteria. AB - Removal of heavy metals by an enriched consortium of sulfate-reducing bacteria (SRB) was evaluated through the abundance of SRB, sulfate reduction, sulfide production and heavy metal precipitation. Five parallel anaerobic semi-continuous stirred tank reactors (CSTR, V = 2 L) (referred as R1-R5) were fed with synthetic wastewater containing mixtures of Cu(2+), Zn(2+), Ni(2+), and Cr(6+) in the concentrations of 30, 60, 90, 120, and 150 mg L(-1) of each metal and operated with a hydraulic retention time of 20 days for 12 weeks. The loading rates of each metal in R1-R5 were 1.5, 3, 4.5, 6, and 7.5 mg L(-1) d(-1), respectively. The results showed that there was no inhibition of SRB growth and that heavy metal removal efficiencies of 94-100% for Cu(2+), Zn(2+), Ni(2+), and Cr(6+) were achieved in R1-R3 throughout the experiment and in R4 during the first 8 weeks. The toxic effect of heavy metals on the SRB consortium was revealed in R5, in which no SRB could survive and almost no heavy metal precipitation was detected after four weeks of operation. PMID- 21632087 TI - Assessment of nitrification in groundwater filters for drinking water production by qPCR and activity measurement. AB - In groundwater treatment for drinking water production, the causes of nitrification problems and the effectiveness of process optimization in rapid sand filters are often not clear. To assess both issues, the performance of a full-scale groundwater filter with nitrification problems and another filter with complete nitrification and pretreatment by subsurface aeration was monitored over nine months. Quantitative real-time polymerase chain reaction (qPCR) targeting the amoA gene of bacteria and archaea and activity measurements of ammonia oxidation were used to regularly evaluate water and filter sand samples. Results demonstrated that subsurface aeration stimulated the growth of ammonia-oxidizing prokaryotes (AOP) in the aquifer. Cell balances, using qPCR counts of AOP for each filter, showed that the inoculated AOP numbers from the aquifer were marginal compared with AOP numbers detected in the filter. Excessive washout of AOP was not observed and did not cause the nitrification problems. Ammonia oxidizing archaea grew in both filters, but only in low numbers compared to bacteria. The cell-specific nitrification rate in the sand and backwash water samples was high for the subsurface aerated filter, but systematically much lower for the filter with nitrification problems. From this, we conclude that incomplete nitrification was caused by nutrient limitation. PMID- 21632088 TI - Assessment of short and long-term effects of imidacloprid on the burrowing behaviour of two earthworm species (Aporrectodea caliginosa and Lumbricus terrestris) by using 2D and 3D post-exposure techniques. AB - Adverse effects of agrochemicals on earthworms' burrowing behaviour can have crucial impacts on the entire ecosystem. In the present study, we have therefore assessed short- and long-term effects on burrowing behaviour in the earthworm species Aporrectodea caliginosa and Lumbricus terrestris after exposure to a range of imidacloprid concentrations (0.2-4 mg kg(-1) dry weight (DW)) for different exposure times (1, 7, 14 d). 2D-terraria were used for the examination of post-exposure short-term effects (24-96 h), while post-exposure long-term effects were assessed by means of X-ray burrow reconstruction in three dimensional soil cores (6 weeks). For the latter each core was incubated with two specimens of L. terrestris and four of A. calignosa. Short-term effects on the burrowing behaviour (2D) of A. caliginosa were already detected at the lowest test concentration (0.2 mg kg(-1) DW), whereas such effects in L. terrestris were not observed until exposure to concentrations 10 times higher (2 mg kg(-1) DW). For both species tested in the 2D-terraria, "total burrow length after 24 h" and "maximal burrow depth after 24 h" were the most sensitive endpoints. 3D reconstructions of the burrow systems made by both earthworm species in the repacked soil cores revealed a significant linear decrease in burrow volume with increasing imidacloprid concentration. Since many of the observed effects occurred at imidacloprid concentrations relevant to natural conditions and since reduced activities of earthworms in soils can have crucial impacts on the ecosystem level, our results are of environmental concern. PMID- 21632089 TI - Per- and polyfluorinated compounds in house dust and indoor air from northern Norway - a pilot study. AB - Polyfluorinated compounds (PFCs) are an extremely versatile class of compounds and are used in a variety of consumer applications and products. Recent studies have suggested that PFCs in indoor air and dust could act as sources of human exposure and outdoor air contamination. This study presents method development and analysis of a wide range of PFCs in dust and air using active sampling techniques with commercially available sampling equipment (forensic nozzles with filter housings for dust collection and polyurethane foam (PUF)-XAD(2)-PUF sandwich-tubes for air sampling) using both liquid and gas chromatography mass spectrometry. The developed method was validated and tested for applicability to analyze dust and air samples at both low and high concentrations (0.5 ng and 25 ng per analyte per air sample, respectively). Samples from private households and an office building were analyzed to explore differences in distribution patterns and concentrations. Perfluorooctane sulfonate, perfluorodecane sulfonate, perfluoroheptanoate, perfluorooctanoate and perfluorononanoate were observed in all samples of dust from private households, in the range from 1 to 80.1 ng g( 1). Fluorotelomer alcohols (FTOHs) were the predominant PFCs in indoor air samples with ?FTOHs ranging between 4.7 and 17.9 ng m(-3). The concentrations found in the present study are generally lower than those previously reported. This variation may be due to differences associated with geographical locations and lifestyles. However, use of different sampling techniques and strategies among studies can introduce large variations in PFC concentration found, making direct comparisons challenging. PMID- 21632090 TI - Impact of model uncertainty on soil quality standards for cadmium in rice paddy fields. AB - At present, soil quality standards used for agriculture do not consider the influence of pH and CEC on the uptake of pollutants by crops. A database with 750 selected paired samples of cadmium (Cd) in soil and paddy rice was used to calibrate soil to plant transfer models using the soil metal content, pH, and CEC or soil Cd and Zn extracted by 0.01 M CaCl2 as explanatory variables. The models were validated against a set of 2300 data points not used in the calibration. These models were then used inversely to derive soil quality standards for Japonica and Indica rice cultivars based on the food quality standards for rice. To account for model uncertainty, strict soil quality standards were derived considering a maximum probability that rice exceeds the food quality standard equal to 10 or 5%. Model derived soil standards based on Aqua Regia ranged from less than 0.3 mg kg-1 for Indica at pH 4.5 to more than 6 mg kg-1 for Japonica type cultivars in clay soils at pH 7. Based on the CaCl2 extract, standards ranged from 0.03 mg kg-1 Cd for Indica cultivars to 0.1 mg kg-1 Cd for Japonica cultivars. For both Japonica and Indica-type cultivars, the soil quality standards must be reduced by a factor of 2 to 3 to obtain the strict standards. The strong impact of pH and CEC on soil quality standards implies that it is essential to correct for soil type when deriving national or local standards. Validation on the remaining 2300 samples indicated that both types of models were able to accurately predict (> 92%) whether rice grown on a specific soil will meet the food quality standard used in Taiwan. PMID- 21632091 TI - Assessing remotely sensed chlorophyll-a for the implementation of the Water Framework Directive in European perialpine lakes. AB - The lakes of the European perialpine region constitute a large water reservoir, which is threatened by the anthropogenic pressure altering water quality. The Water Framework Directive of the European Commission aims to protect water resources and monitoring is seen as an essential step for achieving this goal. Remote sensing can provide frequent data for large scale studies of water quality parameters such as chlorophyll-a (chl-a). In this work we use a dataset of maps of chl-a derived from over 200 MERIS (MEdium Resolution Imaging Spectrometer) satellite images for comparing water quality of 12 perialpine lakes in the period 2003-2009. Besides the different trophic levels of the lakes, results confirm that the seasonal variability of chl-a concentration is particularly pronounced during spring and autumn especially for the more eutrophic lakes. We show that relying on only one sample for the assessment of lake water quality during the season might lead to misleading results and erroneous assignments to quality classes. Time series MERIS data represents a suitable and cost-effective technology to fill this gap, depicting the dynamics of the surface waters of lakes in agreement with the evolution of natural phenomena. PMID- 21632092 TI - The impact of hardpans and cemented layers on oxygen diffusivity in mining waste heaps: diffusion experiments and modelling studies. AB - This study reports column tests and modelling results to assess the impact of hardpans and cemented layers on oxygen supply in mine waste sediments. The analysed sediment samples were obtained from a low-sulphide and low-carbonate polymetallic mine waste tailings impoundment located in the Freiberg mining district in Germany. The three samples were characterised by different degrees and types of cementation. After physical and mineralogical properties of the samples had been determined, breakthrough curves of oxygen were measured in column studies at different degrees of water saturation, and the diffusivities were assessed using a numerical modelling approach. Results demonstrate that cemented layers and hardpans in undisturbed sediments associated with fine grained material operate as preferential pathways for diffusive gas transport during rewetting, leading to higher oxygen diffusivities compared to disturbed sediments. Under air-dry conditions, the disturbed samples show higher diffusivities than the undisturbed sample, indicating clogging of the porosity by precipitation of secondary minerals such as trivalent Fe oxyhydroxides acting as a barrier and thereby decreasing the diffusivity of the undisturbed sample. In contrast to sediments without cementation, diffusion experiments of sediments with cemented layers used in this study yield similar tortuosities in spite of their different grain size distributions, pointing to the important role of these heterogeneities for gas diffusion. PMID- 21632093 TI - Quantum dot transport in soil, plants, and insects. AB - Environmental risk assessment of nanomaterials requires information not only on their toxicity to non-target organisms, but also on their potential exposure pathways. Here we report on the transport and fate of quantum dots (QDs) in the total environment: from soils, through their uptake into plants, to their passage through insects following ingestion. Our QDs are nanoparticles with an average particle size of 6.5 nm. Breakthrough curves obtained with CdTe/mercaptopropionic acid QDs applied to columns of top soil from a New Zealand organic apple orchard, a Hastings silt loam, showed there to be preferential flow through the soil's macropores. Yet the effluent recovery of QDs was just 60%, even after several pore volumes, indicating that about 40% of the influent QDs were filtered and retained by the soil column via some unknown exchange/adsorption/sequestration mechanism. Glycine-, mercaptosuccinic acid-, cysteine-, and amine-conjugated CdSe/ZnS QDs were visibly transported to a limited extent in the vasculature of ryegrass (Lolium perenne), onion (Allium cepa) and chrysanthemum (Chrysanthemum sp.) plants when cut stems were placed in aqueous QD solutions. However, they were not seen to be taken up at all by rooted whole plants of ryegrass, onion, or Arabidopsis thaliana placed in these solutions. Leafroller (Lepidoptera: Tortricidae) larvae fed with these QDs for two or four days, showed fluorescence along the entire gut, in their frass (larval feces), and, at a lower intensity, in their haemolymph. Fluorescent QDs were also observed and elevated cadmium levels detected inside the bodies of adult moths that had been fed QDs as larvae. These results suggest that exposure scenarios for QDs in the total environment could be quite complex and variable in each environmental domain. PMID- 21632094 TI - Chemical characterization of PM10 and PM2.5 mass concentrations emitted by heterogeneous traffic. AB - In this paper, the chemical characterization of PM10 and PM2.5 mass concentrations emitted by heterogeneous traffic in Chennai city during monsoon, winter and summer seasons were analysed. The 24-h averages of PM10 and PM2.5 mass concentrations, showed higher concentrations during the winter season (PM10=98 MUg/m3; PM2.5=74 MUg/m3) followed by the monsoon (PM10=87 MUg/m3; PM2.5=56 MUg/m3) and summer (PM10=77 MUg/m3; PM2.5=67 MUg/m3) seasons. The assessment of 24-h average PM10 and PM2.5 concentrations was indicated as violation of the world health organization (WHO standard for PM10=50 MUg/m3 and PM2.5=25 MUg/m3) and Indian national ambient air quality standards (NAAQS for PM10=100 MUg/m3 and PM2.5=60 MUg/m3). The chemicals characterization of PM10 and PM2.5 samples (22 samples) for each season were made for water soluble ions using Ion Chromatography (IC) and trace metals by Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES) instrument. Results showed the dominance of crustal elements (Ca, Mg, Al, Fe and K), followed by marine aerosols (Na and K) and trace elements (Zn, Ba, Be, Ca, Cd, Co, Cr, Cu, Mn, Ni, Pb, Se, Sr and Te) emitted from road traffic in both PM10 and PM2.5 mass. The ionic species concentration in PM10 and PM2.5 mass consists of 47-65% of anions and 35-53% of cations with dominance of SO42- ions. Comparison of the metallic and ionic species in PM10 and PM2.5 mass indicated the contributions from sea and crustal soil emissions to the coarse particles and traffic emissions to fine particles. PMID- 21632095 TI - Impact of population and latrines on fecal contamination of ponds in rural Bangladesh. AB - A majority of households in Bangladesh rely on pond water for hygiene. Exposure to pond water fecal contamination could therefore still contribute to diarrheal disease despite the installation of numerous tubewells for drinking. The objectives of this study are to determine the predominant sources (human or livestock) of fecal pollution in ponds and examine the association between local population, latrine density, latrine quality and concentrations of fecal bacteria and pathogens in pond water. Forty-three ponds were analyzed for E. coli using culture-based methods and E. coli, Bacteroidales and adenovirus using quantitative PCR. Population and sanitation spatial data were collected and measured against pond fecal contamination. Humans were the dominant source of fecal contamination in 79% of the ponds according to Bacteroidales measurements. Ponds directly receiving latrine effluent had the highest concentrations of fecal indicator bacteria (up to 106 Most Probable Number (MPN) of culturable E. coli per 100 mL). Concentrations of fecal indicator bacteria correlated with population surveyed within a distance of 30-70 m (p<0.05) and total latrines surveyed within 50-70 m (p<0.05). Unsanitary latrines (visible effluent or open pits) within the pond drainage basin were also significantly correlated to fecal indicator concentrations (p<0.05). Water in the vast majority of the surveyed ponds contained unsafe levels of fecal contamination attributable primarily to unsanitary latrines, and to lesser extent, to sanitary latrines and cattle. Since the majority of fecal pollution is derived from human waste, continued use of pond water could help explain the persistence of diarrheal disease in rural South Asia. PMID- 21632096 TI - No association between the ITGA2 807T allele and retinopathy in french patients with type 2 diabetes. PMID- 21632097 TI - Feasibility of laparoscopic extraperitoneal pelvic lymphadenectomy in gynecologic malignancies. AB - OBJECTIVE: The aim of this study is to evaluate the feasibility of laparoscopic extraperitoneal pelvic lymphadenectomy (LEPL) in gynecologic malignancies. METHODS: Twenty-nine women with cervical, ovarian or endometrial cancer underwent laparoscopic extraperitoneal pelvic lymphadenectomy between July 2008 and December 2010. The operating time, nodal yield, blood loss and complications were recorded. RESULTS: The number of patients with cervical, ovarian and endometrial carcinoma was 14, 3 and 12, respectively. The median age of patients was 48.9+/ 12.6 years. The median body mass index was 25.6+/-4.8. Conversion to the transperitoneal laparoscopic approach was necessary in 6 patients for peritoneal tears causing CO(2) gas leakage. Among the remaining 23 patients, the median operating time for laparoscopic extraperitoneal pelvic lymphadenectomy was 69 min (range 50-126 min), and the median estimated blood loss was 20 ml (range 5-105 ml). The median total number of resected nodes was 26 (range 14-42), and complications related to the procedure were rare. CONCLUSIONS: Laparoscopic extraperitoneal pelvic lymphadenectomy is a feasible and safe procedure. It can be used in gynecologic malignancies. PMID- 21632098 TI - Clinical, colposcopic and pathological characteristics of cervical and vaginal high-grade lesions negative for HPV by Hybrid Capture 2. AB - OBJECTIVE: Less than 5% of women with cervical or vaginal biopsy proven high grade squamous intraepithelial lesions (HG-SIL) show a negative Hybrid Capture 2 (HC2) result. We analyzed 1) human papillomavirus (HPV) genotypes by PCR in order to determine whether these cases represent infections by common or unusual types, and 2) the clinical, colposcopic and pathological differential characteristics of these patients. METHODS: 646 women with a histological diagnosis of HG-SIL and a HC2 test collected within 6 months prior to the diagnosis were identified. Patients with a negative HC2 result were selected. HPV was typed in the biopsy specimen in all by PCR using SPF10 and GP5+/6+ primers, and p16(INK4a) immunostaining was performed. The clinical and colposcopy findings of these women were compared with a control group of HG-SIL with positive HC2 result. RESULTS: 20 women (3.1%) with HG-SIL had a negative HC2. All biopsies were positive for p16(INK4). PCR analysis detected HPV types included in HC2 test in 55% of the cases, with an identical percentage of common viruses between women with relative light unit values above or below 0.40 (p=.361). False negative HC2 tests increased with age (p=.002) and were more frequent in patients with non satisfactory colposcopy or small sized lesions (p<.001). CONCLUSION: A negative HC2 test is an infrequent event in women with HG-SIL. Common HPV types are identified in over half of the cases. Older women and patients with small lesions or non satisfactory colposcopy have a higher rate of HC2 negative results. PMID- 21632099 TI - Analysis of racial disparities in stage IIIC epithelial ovarian cancer care and outcomes in a tertiary gynecologic oncology referral center. AB - OBJECTIVE: To examine disparities in delivery of care and survival according to racial classification among White and African-American women with Stage IIIC epithelial ovarian cancer undergoing initial treatment in a tertiary referral center setting. METHODS: All consecutive patients diagnosed with Stage IIIC epithelial ovarian cancer between 1/1/95 and 12/31/08 were identified and clinic pathologic variables retrospectively collected. Differences in initial treatment paradigm, surgical and adjuvant therapy, and overall survival according to racial classification were assessed by univariate and multivariate analyses. RESULTS: A total of 405 patients (White, n=366; African-American, n=39) were identified. There were no significant differences according to racial classification in age, CA125, ASA class, histology, tumor grade, the frequency of initial surgery (90.4% vs 82.1%, p=0.06), optimal residual disease (73.0% vs 69.2%, p=0.28), no gross residual disease (51.4% vs 53.8%, p=0.49), and platinum-taxane chemotherapy (88.3% vs 87.2%, p=0.55). The median overall survival for White patients was 50.5 months (95%CI=43.2-57.9 months), compared to 47.0 (95%CI=36.2-57.8) months for African-Americans (p=0.57). On multivariate analysis, age, tumor grade 3, serum albumin <3.0 g/dl, platinum-based chemotherapy, and no gross residual disease were independently associated with overall survival, while African-American race was not (HR=1.06, 95%CI=0.61-1.79). CONCLUSIONS: Among women undergoing initial treatment for ovarian cancer at a tertiary referral center, African-American patients were as likely as White patients to undergo cytoreductive surgery, be left with minimal post-surgical residual disease, and receive appropriate chemotherapy. With equal access to gynecologic oncology care and multidisciplinary cancer resources, the survival disparities according to race observed in population-based studies are largely mitigated. PMID- 21632100 TI - Correlation between LRIG1 and LRIG2 expressions and expression of 11 tumor markers, with special reference to tumor suppressors, in CIN and normal cervical epithelium. AB - OBJECTIVE: Novel biological markers LRIG1 and LRIG2 have been associated with favorable as well as poor prognosis, respectively, in different cancer types, including cervical cancer. The aim of this study was to investigate possible interactions between these proteins and other tumor markers, and as diagnostic adjuncts in CIN. METHODS: Cervical biopsies from 171 women, with normal epithelium, and low-grade and high-grade CIN were stained for LRIG1 and LRIG2, and 11 additional tumor markers. The tumor markers were chosen to be relevant in cervical neoplasms. Staining was evaluated semiquantitatively. RESULTS: Expression of LRIG1 and LRIG2 was found to correlate with increasing CIN grade, as well as with expression of tumor suppressor FHIT, independent of histological grade. In addition, tumor promoter LRIG2 expression correlated negatively with expression of tumor suppressor retinoblastoma protein and positively with IL-10. The latter correlation did not however remain after adjustment for CIN grade. p53 and p16 expressions correlated positively with LRIG1 expression in univariate analyses, but significance did not hold after adjustment for CIN grade. CONCLUSION: LRIG1 and LRIG2 expressions were seen in precancerous cervical epithelium and found to increase with increasing grade. There was an association between expression of these glycoproteins and FHIT tumor suppressor protein, independently of histological grade. PMID- 21632101 TI - Trace metals (Cd, Cr, Cu, Fe, Ni, Pb, and Zn) in feathers of Black-browed Albatross Thalassarche melanophrys attending the Patagonian Shelf. AB - We investigated the concentrations of cadmium, chromium, copper, iron, nickel, lead and zinc among feather tissues in sexes of Black-browed Albatross Thalassarche melanophrys killed in longliners off Argentina in 2005. We found no different metal concentration with sex for cadmium, copper, iron, lead and zinc in feathers of adult birds, though there were significant body-size differences between sexes. However, the concentrations of trace metals differed significantly among the type of feather within individual bird. The mean concentrations of copper, iron, and zinc in breast feathers of T. melanophrys were lower than those reported for the species from Georgias del Sur/South Georgia, the southern Indian Ocean and for other seabirds' worldwide. While cadmium fall within the known range of concentrations for bird feathers lead were not. Our results may be indicating that level of pollution in Patagonia may not be as negligible as previously thought at least for some trace metals. PMID- 21632102 TI - Metabolic responses in gills of Manila clam Ruditapes philippinarum exposed to copper using NMR-based metabolomics. AB - Copper is an important heavy metal contaminant with high ecological risk in the Bohai Sea. In this study, the metabolic responses in the bioindicator, Manila clam (Ruditapes philippinarum), to the environmentally relevant copper exposures were characterized using NMR-based metabolomics. The significant metabolic changes corresponding to copper exposures were related to osmolytes, intermediates of the Krebs cycle and amino acids, such as the increase in homarine, branched chain amino acids and decrease in succinate, alanine and dimethylamine in the copper-exposed clam gills during 96 h exposure period. Overall, Cu may lead to the disturbances in osmotic regulation and energy metabolism in clams during 96 h experimental period. These results demonstrate that NMR-based metabolomics is applicable for the discovery of metabolic biomarkers which could be used to elucidate the toxicological mechanisms of marine heavy metal contaminants. PMID- 21632103 TI - Iron oxide nanoparticle-containing microbubble composites as contrast agents for MR and ultrasound dual-modality imaging. AB - Magnetic resonance (MR) and ultrasound (US) imaging are widely used diagnostic modalities for various experimental and clinical applications. In this study, iron oxide nanoparticle-embedded polymeric microbubbles were designed as multi modal contrast agents for hybrid MR-US imaging. These magnetic nano-in-micro imaging probes were prepared via a one-pot emulsion polymerization to form poly(butyl cyanoacrylate) microbubbles, along with the oil-in-water (O/W) encapsulation of iron oxide nanoparticles in the bubble shell. The nano-in-micro embedding strategy was validated using NMR and electron microscopy. These hybrid imaging agents exhibited strong contrast in US and an increased transversal relaxation rate in MR. Moreover, a significant increase in longitudinal and transversal relaxivities was observed after US-induced bubble destruction, which demonstrated triggerable MR imaging properties. Proof-of-principle in vivo experiments confirmed that these nanoparticle-embedded microbubble composites are suitable contrast agents for both MR and US imaging. In summary, these magnetic nano-in-micro hybrid materials are highly interesting systems for bimodal MR-US imaging, and their enhanced relaxivities upon US-induced destruction recommend them as potential vehicles for MR-guided US-mediated drug and gene delivery. PMID- 21632104 TI - Magnetically-enabled and MR-monitored selective brain tumor protein delivery in rats via magnetic nanocarriers. AB - The delivery of bioactive proteins to tumors is associated with many difficulties that have impeded clinical translation of these promising therapeutics. Herein we present an approach, including (1) use of magnetically-responsive and MRI-visible nanoparticles as drug carriers, (2) topography-optimized intra-arterial magnetic targeting, (3) MRI-guided subject alignment within the magnetic field, and (4) surface modification of the protein drug with membrane-permeable polyethyleneimine (PEI), to prevail over the obstacles in protein delivery. Applying these methodologies, we demonstrated the delivery of a significant quantity of beta-galactosidase selectively into brain tumors of glioma-bearing rats, while limiting the exposure of normal brain regions. Clinical viability of the technologies utilized, and the ability to deliver proteins at high nanomolar range tumor concentrations, sufficient to completely eradicate a tumor lesion with existing picomolar-potency protein toxins, renders the prospect of enabling protein-based cancer therapy extremely promising. PMID- 21632105 TI - Creation of bony microenvironment with CaP and cell-derived ECM to enhance human bone-marrow MSC behavior and delivery of BMP-2. AB - Extracellular matrix (ECM) comprises a rich meshwork of proteins and proteoglycans, which not only contains biological cues for cell behavior, but is also a reservoir for binding growth factors and controlling their release. Here we aimed to create a suitable bony microenvironment with cell-derived ECM and biodegradable beta-tricalcium phosphate (beta-TCP). More specifically, we investigated whether the ECM produced by bone marrow-derived mesenchymal stem cells (hBMSC) on a beta-TCP scaffold can bind bone morphogenetic protein-2 (BMP 2) and control its release in a sustained manner, and further examined the effect of ECM and the BMP-2 released from ECM on cell behaviors. The ECM was obtained through culturing the hBMSC on a beta-TCP porous scaffold and performing decellularization and sterilization. SEM, XPS, FTIR, and immunofluorescent staining results indicated the presence of ECM on the beta-TCP and the amount of ECM increased with the incubation time. BMP-2 was loaded onto the beta-TCP with and without ECM by immersing the scaffolds in the BMP-2 solution. The loading and release kinetics of the BMP-2 on the beta-TCP/ECM were significantly slower than those on the beta-TCP. The beta-TCP/ECM exhibited a sustained release profile of the BMP-2, which was also affected by the amount of ECM. This is probably because the beta-TCP/ECM has different binding mechanisms with BMP-2. The beta-TCP/ECM promoted cell proliferation. Furthermore, the BMP-2-loaded beta-TCP/ECM stimulated reorganization of the actin cytoskeleton, increased expression of alkaline phosphatase and calcium deposition by the cells compared to those without BMP-2 loading and the beta-TCP with BMP-2 loading. PMID- 21632106 TI - Natural hemozoin stimulates syncytiotrophoblast to secrete chemokines and recruit peripheral blood mononuclear cells. AB - BACKGROUND: Placental malaria is associated with local accumulation of parasitized erythrocytes, deposition of the parasite hemoglobin metabolite, hemozoin, and accumulation of mononuclear cells in the intervillous space. Fetal syncytiotrophoblast cells in contact with maternal blood are known to respond immunologically to cytoadherent Plasmodium falciparum-infected erythrocytes, but their responsiveness to hemozoin, a potent pro-inflammatory stimulator of monocytes, macrophages and dendritic cells, is not known. METHODS: The biochemical and immunological changes induced in primary syncytiotrophoblast by natural hemozoin was assessed. Changes in syncytiotrophoblast mitogen-activated protein kinase activation was assessed by immunoblotting and secreted cytokine and chemokine proteins were assayed by ELISA. Chemotaxis of peripheral blood mononuclear cells was assessed using a two-chamber assay system and flow cytometry was used to assess the activation of primary monocytes by hemozoin stimulated syncytiotrophoblast conditioned medium. RESULTS: Hemozoin stimulation induced ERK1/2 phosphorylation. Treated cells secreted CXCL8, CCL3, CCL4, and tumor necrosis factor and released soluble intercellular adhesion molecule-1. Furthermore, the dependence of the hemozoin responses on ERK1/2 stimulation was confirmed by inhibition of chemokine release in syncytiotrophoblast treated with an ERK pathway inhibitor. Hemozoin-stimulated cells elicited the specific migration of PBMCs, and conditioned medium from the cells induced the upregulation of intercellular adhesion molecule-1 on primary monocytes. CONCLUSIONS: These findings confirm an immunostimulatory role for hemozoin and expand the cell types known to be responsive to hemozoin to include fetal syncytiotrophoblast. The results provide further evidence that syncytiotrophoblast cells can influence the local maternal immune response to placental malaria. PMID- 21632107 TI - Subcutaneous panniculitis-like T-cell lymphoma-associated hemophagocytic syndrome with fulminant relapse during corticosteroid treatment and fatal outcome. PMID- 21632108 TI - Heat shock protein 90 inhibition results in altered downstream signaling of mutant KIT and exerts synergistic effects on Kasumi-1 cells when combining with histone deacetylase inhibitor. AB - KIT mutations may be associated with a poor prognosis in t(8;21) AML. Heat shock protein 90 (Hsp90) is a molecular chaperone frequently used by cancer cells to stabilize mutant oncoproteins. Inhibition of Hsp90 by 17-allylamino-17 demethoxygeldanamycin (17-AAG) disrupted downstream signaling pathways of mutant KIT in Kasumi-1 cells. AML1-ETO fusion gene and mutated KIT act as "two-hit" factors in Kasumi-1 cells. Histone deacetylation (HDAC) inhibitors sodium phenylbutyrate (PB) and valproic acid (VPA) block AML1-ETO. Co-treatment with 17 AAG and PB or 17-AAG and VPA resulted in a synergistic effect in Kasumi-1 cells. Our results confirmed that Hsp90 and mutated KIT were valid molecular targets in the therapy of AML. PMID- 21632109 TI - Changes in parental depression symptoms during family preservation services. AB - OBJECTIVES: Parental depression symptoms often change over the course of child welfare family preservation and parenting services. This raises the question of whether certain processes in family preservation services might be associated with depression symptom change. This study tests three correlational models of change among family preservation service participants: (a) changes in depression symptoms are one facet of broad general changes in wellbeing; (b) the quality of the home visitor-client relationship is associated depression symptom changes; and (c) linking parents to adjunctive services is associated with symptom changes. METHODS: Participants were 2,175 parents in family preservation services, largely for child neglect, who were surveyed using standard measures at pre-treatment, post-treatment and 6 month follow-up. Change patterns were evaluated using growth models, including bivariate parallel and multivariate second-order models. RESULTS: Parallel growth was noted among depression symptoms and changes in social, economic, familial, and parenting domains. A second order change model positing a global change pattern fit the data well. Working alliance had a modest association with improvement, but successful linkage to outside mental health services was not associated with improvement. CONCLUSIONS: Changes in diverse indicators of wellbeing follow a global pattern which might support use of less complex rather than more fully comprehensive service plans. Findings about lack of adjunctive usual care mental health service benefit may be related to uncontrolled factors and this is a topic in need of additional study. PMID- 21632110 TI - Inorganic arsenic in Chinese food and its cancer risk. AB - Even moderate arsenic exposure may lead to health problems, and thus quantifying inorganic arsenic (iAs) exposure from food for different population groups in China is essential. By analyzing the data from the China National Nutrition and Health Survey (CNNHS) and collecting reported values of iAs in major food groups, we developed a framework of calculating average iAs daily intake for different regions of China. Based on this framework, cancer risks from iAs in food was deterministically and probabilistically quantified. The article presents estimates for health risk due to the ingestion of food products contaminated with arsenic. Both per individual and for total population estimates were obtained. For the total population, daily iAs intake is around 42 MUg day(-1), and rice is the largest contributor of total iAs intake accounting for about 60%. Incremental lifetime cancer risk from food iAs intake is 106 per 100,000 for adult individuals and the median population cancer risk is 177 per 100,000 varying between regions. Population in the Southern region has a higher cancer risk than that in the Northern region and the total population. Sensitive analysis indicated that cancer slope factor, ingestion rates of rice, aquatic products and iAs concentration in rice were the most relevant variables in the model, as indicated by their higher contribution to variance of the incremental lifetime cancer risk. We conclude that rice may be the largest contributor of iAs through food route for the Chinese people. The population from the South has greater cancer risk than that from the North and the whole population. PMID- 21632111 TI - Characterization of IgA and IgM binding and internalization by surface-expressed human Fcalpha/MU receptor. AB - The Fcalpha/MU receptor (Fcalpha/MUR) is an unusual Fc receptor in that it binds to two different antibody isotypes, IgA and IgM. This receptor is of interest because it is thought to be involved in the capture of IgA- and IgM-immune complexes and antigen presentation. To further characterize this receptor, we were able to stably express human Fcalpha/MUR on the surface of the 293T cell line. Using this system, we determined the affinity of the interactions of the receptor with IgA and IgM, which led to novel insights including the important finding that IgM polymers can bind to human Fcalpha/MUR in the absence of J chain. This is in contrast to the polymeric immunoglobulin receptor (pIgR), which requires the presence of J chain to bind to polymeric IgA and IgM. The dissociation constants (K(d)) of all of the different human IgA isotypes and allotypes for human Fcalpha/MUR were determined, and we show that the N-linked glycans on IgA1 are not required for binding to the receptor. In addition, we demonstrate that IgA can be rapidly internalized by human Fcalpha/MUR in the presence of cross-linking antibody. PMID- 21632112 TI - Interactions of dendritic cells with cancer cells and modulation of surface molecules affect functional properties of CD8+ T cells. AB - To understand the interaction of dendritic cells (DCs) with cancer cells, we investigated molecular changes in DCs following co-culture with cancer cells. DCs co-cultured with Jurkat cancer cells showed remarkable down-regulation of MHC class I molecules, while DCs co-cultured with MCF-7 cancer cells showed minimal changes. Interestingly, down-regulation of MHC class I on DCs was not observed upon treatment with Jurkat cell lysate or culture supernatant, suggesting the importance of direct cell-cell interactions. The expressions of CD40, CD80, CD83, MHC class II, and IL-12p40 on DCs co-cultured with Jurkat cells were only slightly affected. In contrast, DCs co-cultured with MCF-7 cells showed increased expressions of CD80, CD83, CD86, and IL-12p40. Furthermore, DCs co-cultured with Jurkat cells showed a down-regulation of low molecular weight polypeptides (LMP) 7, and of transporter associated with antigen processing (TAP) 1 and 2 at the mRNA expression level. LMP7, TAP2 and beta2-microglobulin (beta2M) were also down regulated at the protein level. We further demonstrated how altered expression of MHC class I on DCs caused by co-culture with cancer cells affected autologous CD8(+) T cells, using the model MHC class I-presented HSV antigen. We found that DCs that had been HSV-treated and co-cultured with Jurkat cells showed a reduced potency to activate CD8(+) T cells. In contrast, HSV-treated DCs that had been co cultured with MCF-7 cells induced activation of CD8(+) T cells, including high expression of CD25, CD69, granzyme B and cytokines, TNF-alpha and IFN-gamma. PMID- 21632113 TI - The mouse small ubiquitin-like modifier-2 (SUMO-2) inhibits interleukin-12 (IL 12) production in mature dendritic cells by blocking the translocation of the p65 subunit of NFkappaB into the nucleus. AB - Post-translational modification by small ubiquitin-like modifier (SUMO) is involved in several significant cellular events. In particular, SUMO-1 and SUMO-4 modifications of IkappaBalpha have been shown to be actively involved in NFkappaB regulation. However, among the SUMO family, the specific function of SUMO-2/3 remains relatively unknown. In addition, it is not clear whether SUMO-2/3 follows the same functional role as SUMO-1 and SUMO-4 during the activation of NFkappaB. In this study, we examined the influence of mouse SUMO-2 during the maturation of dendritic cells (DCs). Our results showed that the ectopic expression of SUMO-2 does not affect the cell surface expression of MHC class II molecule (A(b)) and co-stimulatory molecules (CD80 and CD86), and the efficiency of antigen uptake. However, the ectopic expression of mouse SUMO-2 inhibited IL-12 secretion by blocking the translocation of the p65 subunit of NFkappaB into the nucleus, which led to the polarization of naive CD4(+) T cells to T helper 2 (Th2) shift in vitro. Further analyses showed that SUMO-2 directly modified IkappaBalpha. These results indicate that the functional role of SUMO-2/3 in the regulation of NFkappaB activity was conserved during evolution. PMID- 21632114 TI - Characterization of the 41kDa allergen Asp v 13, a subtilisin-like serine protease from Aspergillus versicolor. AB - Aspergillus versicolor is common on moldy building materials. Asp v 13, the principal allergen is produced by strains collected from across Canada. In this paper, we report a 1833bp Asp v 13 open reading frame predicted to encode a protein of 403 amino acids in length with three introns. A BLAST search of Asp v 13, a phylogenic tree calculation and alignment with its homologous proteins from other species indicated that Asp v 13 is a secretory, subtilisin-like serine protease widely distributed in Aspergillus species. His-tagged Asp v 13 was over expressed in Escherichia coli and purified using Ni-NTA columns with a yield of 1mg/L. Based on immuno binding assay of recombinant protein both antibodies developed against the natural protein, and human sera IgE, the recombinant protein was similar to the natural form. Six IgE- and seven IgG-binding epitopes were also identified with selected human sera along the entire amino acid sequence of Asp v 13. Most residues binding these epitopes are exposed on the surface and correspond to charged regions of the molecule. PMID- 21632115 TI - A definition for influenza pandemics based on historical records. AB - OBJECTIVE: To analyse the records of past influenza outbreaks to determine a definition for pandemics. METHODS: Analysis of publications of large outbreaks of influenza which have occurred since 1889/90, and to match the results against the current definitions of an influenza pandemic. RESULTS: According to the general understanding of a pandemic, nine outbreaks of influenza since 1889/90 satisfy the definition; however, for two of these, occurring in 1900 and 1933, the data are limited. The special condition for an influenza pandemic requires, in one definition, that the virus strain responsible could not have arisen from the previous circulating strain by mutation; and in the second, that the new strain be a different subtype to the previously circulating strain. Both these restrictions deny pandemic status to two, and possibly three, influenza outbreaks which were pandemics according to the more general understanding of the term. These observations suggest that a re-evaluation of the criteria which define influenza pandemics should be carried out. CONCLUSION: The contradiction outlined above brings the previous definitions of an influenza pandemic into question; however, this can be resolved by defining an influenza pandemic by the following criteria. Thus, an influenza pandemic arises at a single, specific place and spreads rapidly to involve numerous countries. The haemagglutinin (HA) of the emergent virus does not cross-react serologically with the previously dominant virus strain(s), and there is a significant lack of immunity in the population against the emergent virus. These three criteria are interlinked and can be determined early to alert authorities who could respond appropriately. Other criteria associated with pandemics are necessarily retrospective, although important and valid. The implications of this definition are discussed. PMID- 21632116 TI - The stability of mastery motivation and its relationship with home environment in infants and toddlers. AB - Mastery motivation (intrinsic drives to explore and master one's environment) is a key developmental element. The aims of this study were to investigate (1) the stability of mastery motivation between 2 and 3 years of age for two genders; and (2) the associations between early home environment and toddlers' mastery motivation in children with typical development. METHODS: Data of 102 children developing typically from a birth cohort study at Northern Taiwan were analyzed in two parts: (1) stability part: mastery motivation of children were measured at 2 and 3 years of age; (2) environment part: child-parent dyads were assessed from birth, 4 months, 6 months, and 2-3 years of age. Outcomes variables were measured at ages 2 and 3 years by the Dimension of Mastery Questionnaire-17th version. Main predictive variables were measured by Home Observation for Measuring Environment Inventory (HOME) to collect data of the qualities of home environment at 6 months and 2 years; by the Revised Infant Temperament Questionnaire to obtain 4-month activity levels; and by the Comprehensive Developmental Inventory for Infants and Toddlers to obtain 2-year developmental quotient (DQ). RESULTS: There was moderate stability of mastery motivation from 2 to 3 years, and girls' stability was higher than boys'. 6-Month HOME rather than 2-year HOME measures were positively and significantly correlated with instrumental mastery motivation even when controlling for gender, activity level, and DQ. CONCLUSION: Mastery motivation had moderate stability during the toddler period. The quality of home environment in infancy appeared to have a significant impact on toddler's mastery motivation. To promote mastery motivation, caregivers should provide better quality of home environment for infants/toddlers during the very early years. PMID- 21632117 TI - Antidepressant-associated mania or hypomania: a comparison with personality and bipolarity features of bipolar I disorder. AB - BACKGROUND: Hypomania/mania during antidepressant treatment is often neglected by clinicians. There are no specific diagnostic criteria for hypomania and mania associated by antidepressant treatment in the bipolar spectrum. The aim of this study is to compare various characteristics of bipolar I disorder and antidepressant-associated mania. METHOD: In this study, 76 bipolar patients who met DSM-IV criteria for bipolar disorder-type I in remission from mania or depression (Group 1; n = 44) and patients with major depression in remission, who had mania associated by antidepressant treatment (Group 2; n = 32), were admitted. All patients were assessed using the SCID I, Bipolarity Index (BI) and a patient data form. First-degree relatives of all patients were evaluated using the Mood Disorder Questionnaire (MDQ). RESULTS: Sociodemographic features of both groups were similar. The rate of major depression in the relatives of Group 2 was significantly higher than in Group 1. The severity of manic symptoms in Group 2 was significantly lower than in Group 1. Those in Group 2 who were diagnosed with their first episode had atypical depressive features. First-degree relatives of patients in Group 1 had higher positive scores on the MDQ. A statistically significant difference was found between the two groups on all dimensions of the BI except family history. LIMITATIONS: This is a cross-sectional study with a relatively small number of subjects. There is no control group of major depressive patients who did not develop mania during antidepressive treatment. CONCLUSIONS: Our results suggest that antidepressant-associated hypomania/mania could be a different subgroup in the bipolar spectrum. PMID- 21632118 TI - The role of galectin-1 in trophoblast differentiation and signal transduction. AB - Galectins are proteins with the ability to bind beta-galactosides through a conserved carbohydrate recognition domain. Galectin-1 exerts its biological effects by binding glycan ligands on proteins involved in cell adhesion and growth regulation. Galectin-1 inhibits trophoblast cell proliferation and induces syncytium formation. Its down-regulation in the syncytiotrophoblast has been associated with early pregnancy loss. In the choriocarcinoma-derived BeWo cells the galectin-1 induced growth inhibition is apoptosis-independent, but rather appears to be mediated by binding to cell surface receptors, such as the receptor tyrosine kinases REarranged during Transfection (RET) and Janus Kinase (JAK) 2 as well as vascular endothelial growth factor receptor 3. On the syncytiotrophoblast and extravillous trophoblast galectin-1 binds the Thomsen-Friedenreich disaccharide on mucin-1. The cell differentiation processes induced by binding to these receptors ultimately lead to the inhibition of proliferation and syncytium formation. PMID- 21632119 TI - Progesterone-induced blocking factor (PIBF) and trophoblast invasiveness. AB - Controlled trophoblast invasion is a key process during human placentation and a prerequisite for successful pregnancy. Progesterone is one of the factors to regulate trophoblast invasiveness. Progesterone-induced blocking factor (PIBF) is a progesterone-induced molecule expressed by the trophoblast, and also by tumors. The distribution of PIBF within the first-trimester decidua coincides with sites of trophoblast invasion. Another molecule that has been implicated in the control of trophoblast invasiveness is placental leptin. Leptin inhibits the secretion of progesterone by cytotrophoblast. The aim of this work was to investigate the possible interaction of PIBF and leptins in regulating trophoblast invasion. Paraffin-embedded sections from normal first-trimester placentae, partial moles, complete moles, and choriocarcinomas were reacted with PIBF, leptin, and leptin receptor specific antibodies. PIBF-deficient trophoblast cells were generated using siRNA and leptin receptor was detected on Western blot analysis. The lysates of PIBF-treated cells were used for detecting leptin expression in a protein array. PIBF was expressed in both normal first-trimester villous trophoblast and in partial mole. Compared with this, PIBF expression was markedly decreased in complete mole and absent in choriocarcinoma. Neither leptinR nor leptin were detected in partial mole, whereas both of these molecules were present in complete mole and choriocarcinoma. Leptin receptor expression was upregulated in PIBF-deficient cells, while leptin expression was decreased in PIBF-treated cells. These data suggest that PIBF affects the expression of leptin and its receptor, and that PIBF expression is inversely related to trophoblast invasiveness. PMID- 21632120 TI - Uterine and circulating natural killer cells and their roles in women with recurrent pregnancy loss, implantation failure and preeclampsia. AB - The regulation of uterine and circulating peripheral blood natural killer (NK) cells has been associated with reproductive conditions including recurrent pregnancy loss (RPL), implantation failure and preeclampsia. Natural cytotoxicity receptors (NCRs) are unique markers that regulate NK cell cytotoxicity and cytokine production. The role of NCRs in reproductive events has not yet been fully characterized. There is an NK1 (Type 1) shift in peripheral blood NK cells in non-pregnant women prone to RPL and implantation failure. The different profile of NCR expression in endometrial or aborted decidual NK cells suggests the presence of abnormal regulation of NK cells in women with reproductive failure. Women with a history of RPL and preeclampsia carry immunological abnormalities of NCRs on peripheral blood NK cells during pregnancy. Evaluation of NKp46 on peripheral blood NK cells may be applicable for the prediction of preeclampsia. The lower expression of NKp46(+) NK cells in women with preeclampsia may account for the higher production of NK1 cytokines - known as the NK1 shift - in pregnant women with preeclampsia. In this review, the expression of NCRs in peripheral blood NK cells and endometrial or decidual NK cells is discussed in relation to reproductive failure. PMID- 21632121 TI - Blastocyst implantation is vulnerable to stress-induced rises in endogenous estrogens and also to excretions of estrogens by proximate males. AB - Although estrogens help to prepare the uterus for blastocyst implantation, small elevations above optimal levels can prevent implantation. In diverse mammals, stressors including extreme temperatures, physical restraint, environment changes, and predator exposure can impede implantation. This can be mimicked by treating inseminated females with exogenous estrogens. Peri-implantation stressors can elevate estradiol levels, while exogenous estrogen antibodies can mitigate the influences of stress on implantation. Another stimulus that disrupts implantation is exposure to novel males (the "Bruce effect"). This effect, best known in mice, is mediated by chemical factors in male urine. Although a longstanding hypothesis relates the Bruce effect to the female's olfactory memory trace of the sire and reaction to odors of novel males, evidence increasingly supports an alternative hypothesis that implicates males' excreted estrogens. Male urine contains substantial amounts of unconjugated estradiol, and males actively deliver urine toward females which impinges on their nasal region. The Bruce effect can be diminished by drug treatment of males that reduces urinary estradiol, and by treatment of females with estrogen antibodies. Tritiated estradiol ((3)H-E(2)) systemically delivered to male mice was evident in their urine. When (3)H-E(2) was given intra-nasally to inseminate females, it was found in their circulation and in diverse tissues, with greatest radioactivity in the uterus. Accordingly, evidence indicates that males' excreted estradiol can arrive at the female reproductive tract, where it can disrupt implantation through known mechanisms. PMID- 21632122 TI - Minor physical anomalies and neurological soft signs in patients with schizophrenia and their siblings. AB - Neurological soft signs (NSSs) and minor physical anomalies (MPAs) are consistently found at higher rates in individuals with schizophrenia compared to healthy controls. However, limited research has been conducted on these traits among the biological relatives of these patients. We aimed to identify the possible origins of these traits in schizophrenia by exploring them in patients with schizophrenia, their healthy siblings and normal controls. Ninety-six patients with schizophrenia, their 66 non-psychotic siblings and 52 healthy subjects were studied. Measures included the Neurological Evaluation Scale, a structured examination for detection of minor physical anomalies, stroop and verbal fluency tests for cognitive assessment, and scales for assessment of disease severity in patients; the Scale for the Assesment of Negative Symptoms and the Scale for the Assesment of Positive Symptoms. Increased rates of NSSs and high MPA scores were found in both the patients and their siblings as compared to normal controls. MPAs in several body regions were similar (eyes, ears, hands and feet) or correlated (innercanthal width and head circumference) between patients and their respective siblings. However, there was little similarity in palate and tongue anomalies between these subjects. These results suggest that NSSs and MPAs might represent two distinct markers of risk for schizophrenia. MPAs at different locations may also represent distinct pathological processes, such that palate and tongue abnormalities are more likely to represent non-familial rather than familial factors compared to other abnormalities. PMID- 21632123 TI - Impact of the mouth breathing occurred during childhood in the adult age: biophotogrammetric postural analysis. AB - OBJECTIVE: This study aims to evaluate the impact of the mouth breathing occurred during childhood on the body posture in the adult age. METHODS: 24 adults, of both genders, aged from 18 to 30 years old with report of clinical manifestations of mouth breathing during the childhood composed the study group (SG). The control group (CG) was composed by 20 adults in the same age, without any respiratory problem since the childhood up to the present time. All the volunteers underwent a physiotherapeutic evaluation consisted of anamnesis and postural biophotogrammetry (SAPo v 0.68((r))). The comparison between the data of the SG and CG was accomplished by Student's t-test. RESULTS: The biophotogrammetric analysis demonstrated that the SG showed more forward head posture confirmed by the angles A9 (p = 0.0000) and CL (p = 0.0414) and also by the cervical distance (p = 0.0079). Additionally, this group presented a larger angular measure of the lumbar lordosis (p = 0.0141) compared to the CG. CONCLUSION: The results indicate that adults with mouth-breathing childhood have postural alterations, mainly in the head and lumbar column, which keeps for the whole life. PMID- 21632124 TI - Laryngovideostroboscopy in children--diagnostic possibilities and constraints. AB - OBJECTIVE: The aim of this study was to report our experience with laryngovideostroboscopy (LVS) in consecutively examined children patients. METHODS: The study included 150 children (2.5-14 years of age) diagnosed with dysphonia. Patients were divided into three age groups: group I - from 2.5 to 6 years of age (n = 31), group II - from 6 to 10 years of age (n = 73), and group III--from 10 to 14 years of age (n = 46). LVS was performed during the second visit. 3.5mg of dormicum were administered orally 30 min prior to LVS in all children from group I and in some patients belonging to group II. Local anesthesia of the pharyngeal mucosa was not needed in any case. RESULTS: In nine out of 150 children, it was not possible to perform LVS during the first attempt. In eight of these children, the examination was performed successfully during another visit with satisfactory LVS images obtained. LVS revealed soft vocal fold nodules in 85 patients along with other organic changes found in five children: congenital laryngeal web (n = 1), vocal fold cysts (n = 3), and vocal fold paralysis (n = 1). Hyper-functional dysphonia was diagnosed in the remaining 60 subjects. CONCLUSIONS: In order to perform successful LVS in children patient, the purpose and methodology of this examination should be explained to parents on the first visit whereas an attempt to perform LVS should be undertaken during the second visit. Oral administration of dormicum 30min prior to the examination is advisable, particularly in younger children, and allows us to avoid the use of local anesthetics. PMID- 21632125 TI - Review on aging, unloading and reloading: changes in skeletal muscle quantity and quality. AB - Aging and muscle unloading are associated with a decline in muscle mass, structure, force and changes in the mechanical properties of skeletal muscle. There is a perception that reloading restores muscle architecture and function but differences in the regeneration capacity of intra- and extra-cellular compartments, age dependent susceptibility to muscle damage, particularly in the contractile machinery, make it difficult to attribute the main role of the above mentioned components. This review briefly summarizes studies that examine aging skeletal muscle during unloading and reloading. The article highlights changes in muscle mass, structure, extra-cellular matrix (ECM) components, strength, locomotion, adaptability to exercise, peculiarities of protein metabolism, susceptibility to glucocorticoids and recovery of muscle from injury. PMID- 21632126 TI - Life satisfaction (LS) and symptoms among the oldest-old: results from the longitudinal population study called Good Aging in Skane (GAS). AB - Studies on the prevalence of symptoms in the general population and its' relation to LS in the oldest-old are to our knowledge non-existent. The aim of this study was to describe the frequency and experienced severity of elderly subjects' reported symptoms and how symptoms are related to LS. The study population consisted of 681 individuals aged 78-93 years, drawn from the longitudinal population study, GAS, part of a national survey (SNAC). Scores on the life satisfaction index were related to scores on a modified version of the Goteborg Quality of Life (GQoL) instrument, covering 32 common symptoms. Musculo-skeletal symptoms like pain were reported by 74%, 80% had depressive symptoms and 68% general fatigue. Less than 6% of men and women reported no metabolic symptoms or symptoms related to the head. In a multiple regression model it was shown that four groups of symptoms could significantly predict LS 3 years later: depression, tension, GI-symptoms and musculo-skeletal symptoms. The clinical implication of this study is that careful attention should be paid to the elderly patients' complaints concerning symptoms in the above areas since this has the potential to significantly increase the patients' satisfaction with life. PMID- 21632127 TI - Role of cardiovascular risk factors (CRF) in the patients with mild cognitive impairment (MCI). AB - Few therapeutic options are available nowadays to improve the prognosis of patients with Alzheimer's disease (AD). There are rather several evidences in literature that controlling vascular risk factors may be an effective intervention for modifying the course of this disease. The aim of our study was to investigate the role of CRF in 50 patients with MCI according to Petersens's criteria, and to evaluate their influence on cognitive and behavioral features of the disease and on the development of dementia. Statistical analysis of the data showed that the 60% of the patients with MCI and CRF developed dementia, while 40% maintained the same cognitive conditions at the end of the study. Only 32% of the subjects without cardiovascular comorbidities developed dementia. The results of the study suggest that CRF play a key role in cognitive decline of patients with MCI. Patients with MCI and CRF showed not only worse cognitive performances, but also behavioral disorders, depression and functional disability. Patients with CRF had higher conversion rate to AD than the other group, with a mean disease-free period 3 months shorter than the control group. PMID- 21632128 TI - PI3K/Akt signaling pathway involved in cardioprotection of preconditioning with high mobility group box 1 protein during myocardial ischemia and reperfusion. PMID- 21632129 TI - An unusual complication of a Takotsubo cardiomyopathy: a not so benign disease? PMID- 21632130 TI - Diagnostic performance measures of ELISA and quantitative PCR tests for porcine circovirus type 2 exposure using Bayesian latent class analysis. AB - Porcine circovirus 2 (PCV2) is believed to be a necessary but not sufficient underlying cause of porcine circovirus associated disease (PCVAD) in swine (Opriessnig et al., 2007). Since the potential threat of PCVAD is dependent on the prevalence of PCV2 in swine populations, accurate diagnostic tests are important for epidemiologic surveillance. Therefore, we evaluated the diagnostic sensitivity (Se) and specificity (Sp) of a new indirect ELISA and two quantitative PCR tests for PCV2 in a series of latent class models that used Bayesian estimation procedures. A total of 4140 samples from finisher pigs were tested for evidence of PCV2 by the ELISA and a TaqMan (TM) quantitative PCR, 995 by the ELISA and a SYBR Green (SG) dye-binding PCR, 998 by both PCRs and 993 by all three tests. Overall, the median (95% probability interval) ELISA Se and Sp was 0.85 (0.83-0.87) and 0.74 (0.68-0.79), respectively, when all three tests were analyzed together at an ELISA absorbance (optical density or OD) cutoff of >=0.3. The TM PCR Se and Sp was 0.86 (0.84-0.88) and 0.94 (0.87-0.97), respectively, and the SG PCR Se and Sp was 0.83 (0.81-0.85) and 0.98 (0.94-1.00), respectively when all three tests were analyzed together at an ELISA OD cutoff of >=0.3. Sensitivity analysis revealed that Sp estimates in general had less stability than Se estimates, but the SG PCR(Sp) was the most stable. Limited conditional dependence between the two PCR tests was detected. We conclude that the ELISA had the highest diagnostic Se at an absorbance cutoff of >=0.3, while the SG PCR had the highest diagnostic Sp. The prevalence levels for exposure to PCV2 in finishing swine populations across all analyses ranged from 58 to 100%. PMID- 21632131 TI - Soluble microbial products and their implications in mixed culture biotechnology. AB - Soluble microbial products (SMP) are soluble organic compounds released during normal biomass metabolism in mixed culture biotechnology. In this review, we give the up-to-date status on several essential SMP issues: mechanisms of SMP formation, differentiation between utilization-associated products (UAP) and biomass-associated products (BAP), biodegradability of the SMP components, how formation of SMP by autotrophs controls effluent quality and supports a substantial population of heterotrophs, mathematical modeling that includes SMP, and improving effluent quality by controlling SMP. We also present two timely examples that highlight our current understanding and give an indication of how SMP affects the performance of modern mixed culture biotechnology: membrane fouling of membrane bioreactors (MBRs) and the dynamics of SMP in anaerobic systems. PMID- 21632132 TI - The ESTRO Breur Lecture 2010: toward a tailored patient approach in rectal cancer. AB - The last decades have been characterized by tremendous improvements in the treatment of rectal cancer. Based on the evidence gathered in these years, the standard treatment of patients with locally advanced rectal cancer has now become preoperative chemoradiation (CRT) followed by total mesorectal excision. Although the locoregional control with this treatment regimen is quite favorable for the majority of the patients, there is still room for further improvement. For those patients with a good response to preoperative chemoradiation (CRT), extensive surgery could be avoided and replaced by minimal invasive surgery or no surgery at all. To date however, the only way to ascertain a complete remission is pathologic examination of the resection specimen. Early response prediction of the tumor to preoperative CRT is essential for further selection of patients who could be spared invasive surgery. This could be achieved by assessing molecular markers present in the tumor tissue and blood of the patients or by non invasive functional imaging before and during preoperative treatment. For those patients with a less favorable response, treatment intensification might be the way to go. This could be accomplished by dose painting on resistant tumor regions or by the addition of molecular targeted agents to the standard treatment. In this article, we review the current standard of care and the remaining challenges in the treatment of patients with locally advanced rectal cancer. PMID- 21632133 TI - Development of NTCP models for head and neck cancer patients treated with three dimensional conformal radiotherapy for xerostomia and sticky saliva: the role of dosimetric and clinical factors. AB - PURPOSE: The purpose of this multicentre prospective study was to investigate the significance of the radiation dose in the major and minor salivary glands, and other pre-treatment and treatment factors, with regard to the development of patient-rated xerostomia and sticky saliva among head and neck cancer (HNC) patients treated with primary (chemo-) radiotherapy ((CH)RT). METHODS AND MATERIALS: The study population was composed of 167 consecutive HNC patients treated with three-dimensional conformal (3D-CRT) (CH) RT. The primary endpoint was moderate to severe xerostomia (XER6m) as assessed by the EORTC QLQ-H&N35 at 6 months after completing (CH)RT. The secondary endpoint was moderate to severe sticky saliva at 6 months (STIC6m). All organs at risk (OARs) potentially involved in salivary function were delineated on planning-CT, including the parotid, submandibular and sublingual glands and the minor glands in the soft palate, cheeks and lips. Patients with moderate to severe xerostomia or sticky saliva at baseline were excluded. The optimum number of variables for a multivariate logistic regression model was determined using a bootstrapping method. RESULTS: The multivariate analysis showed the mean parotid dose, age and baseline xerostomia (none versus a bit) to be the most important predictors for XER6m. The risk of developing xerostomia increased with age and was higher when minor baseline xerostomia was present in comparison with patients without any xerostomia complaints at baseline. Model performance was good with an area under the curve (AUC) of 0.82. For STIC6m, the mean submandibular dose, age, the mean sublingual dose and baseline sticky saliva (none versus a bit) were most predictive for sticky saliva. The risk of developing STIC6m increased with age and was higher when minor baseline sticky saliva was present in comparison with patients without any sticky saliva complaints at baseline. Model performance was good with an AUC of 0.84. CONCLUSIONS: Dose distributions in the minor salivary glands in patients receiving 3D-CRT have limited significance with regard to patient-rated symptoms related to salivary dysfunction. Besides the parotid and submandibular glands, only the sublingual glands were significantly associated with sticky saliva. In addition, reliable risk estimation also requires information from other factors such as age and baseline subjective scores. When these selected factors are included in predictive models, instead of only dose volume histogram parameters, model performance can be improved significantly. PMID- 21632134 TI - Virus inactivation by salt (NaCl) and phosphate supplemented salt in a 3D collagen matrix model for natural sausage casings. AB - Due to possible presence and spread of contagious animal viruses via natural sausage casings the international trade in these food products is subject to veterinary and public health requirements. In order to manage these restrictions we determined the effect of casing preservation on four highly contagious viruses for livestock: foot-and-mouth-disease virus (FMDV), classical swine fever virus (CSFV), swine vesicular disease virus (SVDV) and African swine fever virus (ASFV). We used an in vitro 3D collagen matrix model in which cells, infected with the four different viruses were embedded in a bovine collagen type I gel matrix and treated with either saturated salt (NaCl) or phosphate supplemented saturated salt at four different temperatures (4, 12, 20 and 25 degrees C) during a period of 30 days. The results showed that all viruses were faster inactivated at higher temperatures, but that stability of the various viruses at 4 degrees C differed. Inactivation of FMDV in the 3D collagen matrix model showed a clear temperature and treatment effect on the reduction of FMDV titres. At 4 and 12 degrees C phosphate supplemented salt showed a very strong FMDV inactivation during the first hour of incubation. Salt (NaCl) only had a minor effect on FMDV inactivation. Phosphate supplemented salt treatment increased the effect temperature had on inactivation of CSFV. In contrast, the salt (NaCl) treatment only increased CSFV inactivation at the higher temperatures (20 degrees C and 25 degrees C). Also SVDV inactivation was increased by phosphate supplemented salt, but salt (NaCl) treatment only resulted in a significant decrease of SVDV titre at a few time points. The ASFV results showed that both salt (NaCl) and phosphate supplemented salt were capable to inactivate ASFV within 48 h. In contrast to the other viruses (FMDV, CSFV and SVDV), ASFV was the most stable virus even at higher temperatures. The results obtained in this in vitro model underline the efficacy of a combined treatment using phosphate supplemented salt and storage at 20 degrees C or higher for a period of 30 days. This treatment may therefore be useful in reducing the animal health risks posed by spread of contagious animal viruses by international trade of natural sausage casings. PMID- 21632135 TI - Survival of Salmonella Newport in oysters. AB - Salmonella enterica is the leading cause of laboratory-confirmed foodborne illness in the United States and raw shellfish consumption is a commonly implicated source of gastrointestinal pathogens. A 2005 epidemiological study done in our laboratory by Brands et al., showed that oysters in the United States are contaminated with Salmonella, and in particular, a specific strain of the Newport serovar. This work sought to further investigate the host-microbe interactions between Salmonella Newport and oysters. A procedure was developed to reliably and repeatedly expose oysters to enteric bacteria and quantify the subsequent levels of bacterial survival. The results show that 10 days after an exposure to Salmonella Newport, an average concentration of 3.7 * 10(3)CFU/g remains within the oyster meat, and even after 60 days there still can be more than 10(2)CFU/g remaining. However, the strain of Newport that predominated in the market survey done by Brands et al. does not survive within oysters or the estuarine environment better than any other strains of Salmonella we tested. Using this same methodology, we compared Salmonella Newport's ability to survive within oysters to a non-pathogenic strain of E. coli and found that after 10 days the concentration of Salmonella was 200-times greater than that of E. coli. We also compared those same strains of Salmonella and E. coli in a depuration process to determine if a constant 120 L/h flux of clean seawater could significantly reduce the concentration of bacteria within oysters and found that after 3 days the oysters retained over 10(4)CFU/g of Salmonella while the oysters exposed to the non-pathogenic strain of E. coli contained 100-times less bacteria. Overall, the results of this study demonstrate that any of the clinically relevant serovars of Salmonella can survive within oysters for significant periods of time after just one exposure event. Based on the drastic differences in survivability between Salmonella and a non-pathogenic relative, the results of this study also suggest that unidentified virulence factors may play a role in Salmonella's interactions with oysters. PMID- 21632136 TI - Chorea-ballism associated with nonketotic hyperglycaemia or diabetic ketoacidosis: characteristics of 25 patients in Korea. AB - Chorea-ballism is a rare form of movement disorder complicated by severe hyperglycaemia and in association with a contralateral basal ganglia lesion. We analysed the clinical characteristics of 25 Korean patients with chorea-ballism associated with nonketotic hyperglycaemia or diabetic ketoacidosis. Possible mechanisms of disease are also discussed. PMID- 21632137 TI - Anniversary of vote on UN Summit: progress to date and challenges remaining. PMID- 21632138 TI - Dorsal versus ventral monofilament testing of the great toe for the identification of diabetic sensorimotor polyneuropathy. AB - We tested whether monofilament examination of the ventral rather than dorsal aspect of the great toe could better detect diabetic sensorimotor polyneuropathy (DSP). Although dorsal testing had better overall performance for identifying Subclinical DSP, the magnitude of benefit was small and there was no difference in identifying Clinical DSP. PMID- 21632139 TI - Diabetic ketoacidosis during gestational diabetes. A case report. AB - Diabetic ketoacidosis is an infrequent complication of gestational diabetes but results in fetal loss. It usually occurs in the later stages of pregnancy. We report two young pregnant women who were admitted because of newly diagnosed diabetes with ketoacidosis. One patient presented with intrauterine fetal demise. PMID- 21632140 TI - Blood glucose and nocturnal blood pressure in African and Caucasian men: The SABPA study. AB - AIM: To investigate the relationship between nocturnal blood pressure and chronically elevated blood glucose to determine if these elevated blood glucose concentrations contribute to a non-dipping blood pressure, especially in high risk groups such as Africans. METHODS: Nocturnal blood pressures and blood glucose levels of 41 non-dipping African and 28 non-dipping Caucasian men were investigated. Ambulatory systolic (SBP) and diastolic blood pressure (DBP) were measured and blood collected in sodium fluoride tubes from the antebrachial vein to determine serum glucose and glycosylated hemoglobin A1c (HbA1c) percentage. The estimated average glucose (eAG) was determined from HbA1c percentage with a regression formula. RESULTS: The African non-dippers had higher blood pressures (p<0.001) and elevated HbA1c (p=0.037) and eAG (p=0.041) levels compared to the Caucasians. In single, partial and multiple regression analyses nighttime (00:00 04:00) SBP correlated positively with HbA1c (p=0.069) and eAG (p<0.001) in the African men. No correlations were found in the Caucasian men. Sensitivity analysis confirmed that the association between nighttime SBP (00:00-04:00) and eAG was independent of carotid intima-media thickness in the African men (R(2)=0.617; beta=0.438; p=0.008). CONCLUSION: The blunted nocturnal decline in SBP during the early morning hours is associated with chronically elevated blood glucose in non-dipper African men. PMID- 21632141 TI - Determination of the cutoff point for waist circumference that establishes the presence of abdominal obesity in Latin American men and women. AB - AIMS: The recent definitions of the metabolic syndrome (MS) recognize the need for ethnic and region-specific waist circumference (WC) cut-offs that identify people with abdominal obesity. We tested WC as a diagnostic tool to identify people with visceral adiposity in Hispanics from the Latin America region. METHODS: We used the area of visceral adipose tissue (VAT) >= 100 cm2 at the level between the 4th and 5th lumbar vertebrae in abdominal CT scans as a marker of visceral adiposity and established the optimal WC threshold for men and women by means of receiver operating characteristic (ROC) curves. RESULTS: 179 men and 278 women from Mexico, El Salvador, Venezuela, Colombia and Paraguay were included. The ROC curves were highly discriminative of excess VAT for men and women (area under the curve 0.9 and 0.8 respectively) and the WC threshold was identified at 94 cm for men and between 90 and 92 cm for women. CONCLUSION: In men the WC cutoff was equal to that proposed for europids and suggested for US adults who may have strong genetic contribution to insulin resistance such as Hispanics. In women the threshold was significantly higher than previously proposed for South and Central Americans. PMID- 21632142 TI - Adipose tissue derived mesenchymal stem cell (AD-MSC) promotes skin wound healing in diabetic rats. AB - AIMS: Stem cells are a new hope to ameliorate impaired diabetic wound healing. The purpose of this study was to evaluate the effect of adipose tissue derived mesenchymal stem cells (AD-MSCs) on wound healing in a diabetic rat model. METHODS: Twenty-six rats became diabetic by a single intraperitoneal injection of streptozotocin. Six rats served as non-diabetic (non-DM). Diabetic rats were divided into two equal groups randomly; control and treatment. Six weeks later, a full-thickness circular excisional wound was created on the dorsum of each rat. AD-MSCs were injected intra-dermally around the wounds of treatment group. PBS was applied to control and non-DM groups. The wound area was measured every other day. After wound healing completion, full thickness skin samples were taken from the wound sites for evaluation of volume density of collagen fibers, length and volume density of vessels, and numerical density of fibroblasts by stereological methods. RESULTS: AD-MSCs accelerated wound healing rate in diabetic rats, but did not increase length and volume density of the vessels and volume density of the collagen fibers. AD-MSCs decreased the numerical density of fibroblasts. CONCLUSIONS: We concluded that AD-MSCs enhances diabetic wound healing rate probably by other mechanisms rather than enhancing angiogenesis or accumulating collagen fibers. PMID- 21632143 TI - Third trimester plasma adiponectin and leptin in gestational diabetes and normal pregnancies. AB - The aim of this study was to compare pregnant women with and without gestational diabetes (GDM) with regard to third trimester adiponectin, leptin and insulin resistance, as well as to investigate their relation to fetal growth and macrosomia. 134 pregnant women were enrolled in the study: 86 with GDM and 48 controls. Maternal plasma adiponectin, leptin, fasting insulin, glucose and fetal biometry were measured between 27 and 32 weeks of gestation. Birthweight and delivery data were also assessed. Adiponectin in GDM patients was lower when compared to non-diabetic women while mean leptin concentration was not different. In GDM group only mothers' weight gain until third trimester was significantly different between the groups of patients with normal and accelerated fetal growth. No correlation of adiponectin and leptin with fetal growth was shown. Mothers' weight gain until third trimester and mean fasting glucose between 33 and 34 weeks of gestation were associated with neonatal macrosomia. There were no differences in adiponectin and leptin between mothers of macrosomic and non macrosomic neonates. Fetal growth seems not to be related to third trimester adiponectin and leptin concentrations, while increased third trimester fasting glucose may be an independent risk factor of macrosomia. PMID- 21632144 TI - Ageing and associations of fasting plasma glucose and 2 h plasma glucose with HbA(1C) in apparently healthy population. "FIN-D2D" study. AB - OBJECTIVE: In this FIN-D2D cross-sectional survey the relationship of age with HbA(1c) and fasting and 2h glucose in the oral glucose tolerance test (OGTT) was explored in apparently randomly selected healthy population. PATIENTS AND METHODS: The glycaemic parameters were measured in 1344 men and 1482 women (aged 45-74 years), and among them we excluded all subjects with known diabetes, hypertension or dyslipidaemia. The final analyses for HbA(1c) and the ratios of fasting glucose/HbA(1c) and 2h glucose/HbA(1c) included 649 men and 804 women. RESULTS: Mean age was 57 years and BMI 26.1kg/m(2) for both genders. HbA(1c) increased in both genders with age (p<0.001). For a particular fasting glucose level HbA(1c) level was higher in older age groups (p<0.001 for linearity). By contrast, a particular 2h plasma glucose value in OGTT implied significantly lower HbA(1c) in the elderly (p<0.001 for linearity). CONCLUSION: In apparently healthy population, screened with OGTT, in older individuals compared with younger ones a particular HbA(1c) value implies slightly lower fasting glucose, but relatively higher 2h glucose. These results need to be verified in different populations. The effects of age on relation between HbA(1c) and plasma glucose should be taken into account in classifying people into different dysglycaemia categories. PMID- 21632145 TI - Mini Nutritional Assessment (MNA) and biochemical markers of cachexia in metastatic lung cancer patients: interrelations and associations with prognosis. AB - PURPOSE: Lung cancer patients frequently present with weight loss in the context of the cachexia syndrome. Despite its high clinical significance, definite diagnostic criteria of cachexia are lacking. Nutritional screening questionnaires, like the Mini Nutritional Assessment (MNA), have been proposed for the timely diagnosis of the syndrome. The aim of this study was to evaluate the correlation of MNA with laboratory markers of inflammation/cachexia in patients with metastatic lung cancer. The prognostic value of the measured parameters was also examined. PATIENTS AND METHODS: Patients with metastatic lung cancer referred for systemic therapy were eligible. Baseline clinical characteristics were recorded and nutritional status was assessed using MNA. Blood samples were also collected and the following parameters were measured: hemoglobin (Hb), albumin (Alb), C-reactive protein (CRP), ghrelin, adiponectin, leptin and insulin growth factor I (IGF-I). RESULTS: Totally, 115 patients (101 males) [median age 66 years (range 32-86)] were evaluated. According to MNA score, 27 (23.5%) patients were well nourished, 59 (51.3%) were at nutritional risk and 29 (25.2%) were already malnourished at diagnosis. MNA correlated with the following parameters: Hb (p=0.001), albumin (p<0.001), CRP (p=0.002), adiponectin (p=0.037) and leptin (p=0.008). After a median follow up of 38.2 months, multivariate analysis revealed that age (p=0.008), number of metastatic sites (p<0.001), MNA (p=0.044) and leptin (p=0.004) independently correlated with overall survival. CONCLUSIONS: Based on the MNA, the majority of patients were either malnourished or at nutritional risk. MNA correlated with laboratory parameters related to inflammation/cachexia and was independently associated with survival. PMID- 21632146 TI - Lighting up the nuclear pore complex. AB - It is generally accepted that transport through the nuclear pore complex (NPC) involves an abundance of phenylalanine-glycine rich protein domains (FG-domains) that serve as docking sites for soluble nuclear transport receptors (NTRs) and their cargo complexes. But the precise mechanism of translocation through the NPC allowing for high speed and selectivity is still vividly debated. To ultimately decipher the underlying gating mechanism it is indispensable to shed more light on the molecular arrangement of FG-domains and the distribution of NTR-binding sites within the central channel of the NPC. In this review we revisit current transport models, summarize recent results regarding translocation through the NPC obtained by super-resolution microscopy and finally discuss the status and potential of optical methods in the analysis of the NPC. PMID- 21632147 TI - Pushed monocanalicular intubation. Pitfalls, deleterious side effects, and complications. AB - PURPOSE: To present our experience with pushed monocanalicular nasolacrimal intubation in the management of 90 consecutive cases of nasolacrimal outflow obstruction. MATERIALS AND METHOD: This paper reports a non-randomized study of 90 consecutive cases treated with a pushed Monoka intubation system (MasterkaTM). A metal guide is placed inside a silicone tube rather than being attached at the distal end of the tube, as done with traditional pulled intubations. Three probe lengths are available: 30, 35, and 40 mm. SURGICAL PROCEDURE: The silicone stent was pushed into a punctum, canaliculus, and nasolacrimal duct by means of the guide. After passing through the valve of Hasner and reaching the nasal floor, the guide was then delicately withdrawn while remaining oriented along the axis of the lacrimal sac and duct. Throughout this phase, the anchoring plug was held in contact with the punctum. Three study groups were set up chronologically: group 1: endo-DCR procedures done with Masterka insertions under endoscopic observation. Group 2: Masterka insertions done with endoscopic guidance. Group 3: blind Masterka insertions without endoscopic guidance. The patients in groups 2 and 3 were selected on the information obtained by lacrimal probing. Only cases with mucosal nasolacrimal stenoses were included. All patients had surgery under general anesthesia with mechanically assisted ventilation (groups 1 and 2) or spontaneous ventilation (group 3). The anchoring plug was inserted into the punctum and vertical canaliculus, either by pulling on the probe (group 1) or using an inserting instrument. RESULTS: A total of 90 pushed Monoka intubations were done. Endoscopic examination (groups 1 and 2) demonstrated visually that the pushed intubation method was effective. In none of the 28 cases did the silicone bunch up when the guide was withdrawn. DEGREE OF DIFFICULTY: This was dependent upon proper selection for pushed Monoka intubation; the length of the probe and confirmation that there no false passage was created. The pushed intubation technique was only slightly more difficult than a simple lacrimal probing. The average operating time, excluding the anesthetic procedures, was respectively 5 min (group 2) and 4 min (group 3). COMPLICATIONS DURING SURGERY: There were no anesthetic or general problems observed in the three groups. Epistaxis was also not noted. POSTOPERATIVE COMPLICATIONS: Fifteen percent (13/90). The 13 complications noted were: two cases of canaliculitis, one intracanalicular migration, eight probes that disappeared, one keratitis, and one case of involuntary removal by the patient. DELETERIOUS SIDE EFFECTS: Tearing with the probe was in place was noted in 21.1% of the cases (19/90). This tearing disappeared as soon as the probe was removed in 50% of these cases (10/19). FUNCTIONAL RESULTS: Overall, the success rate (absence of epiphora, absence of mucous discharge) was 90% (81/90) with an average follow-up period of 19 weeks (Range, 1 day to 60 weeks). Two cases were lost to follow-up at day 1 and day 7. Group 1: 90.9% (20/22 cases; average age: 65 years, with an average follow-up period of 24 weeks). Group 2: 100% (6/6 cases; average age: 3.1 years, with an average follow-up period of 14 weeks). Group 3: 88.3% (53/60 cases excluding the two cases that were lost to follow-up; mean age: 2.3 years, with an average follow-up period of 16 weeks). CONCLUSIONS: From a technical perspective, pushed nasolacrimal intubation is much simpler than the traditional pulled types of nasolacrimal intubation. The anesthetic procedure required is the same as that for a late probing procedure, but the functional results are better. The Masterka is an alternative to simple late probing in the treatment of mucosal nasolacrimal stenoses in patients of over 12 months of age. PMID- 21632148 TI - [Predictive factors for successful Ferrara intracorneal ring segment implantation in keratoconus]. AB - OBJECTIVE: To define predictive factors after manual implantation of intrastromal corneal ring segments (Ferrara ring) for the management of keratoconus. PATIENTS AND METHODS: Twenty-five eyes of 25 patients with keratoconus were implanted with Ferrara intracorneal ring segments using a mechanical procedure. We measured corneal ring implantation depth, best spectacle-corrected visual acuity (BSCVA), as well as refractive and keratometric outcomes. Patients were separated into two groups according to the gain of at least two lines of BSCVA ("good response") or the gain of only one line of BSCVA, no change, or the loss of lines of BSCVA ("poor response"). RESULTS: After a 6-month follow-up, 41% patients experienced a gain of at least two lines of BSCVA, 31.8% a gain of only one line, 13.6% no change, and 13.6% a loss of BSCVA. The sphere reduced from 4.39 to 2.34D (p<0.01), the cylinder from 4.19 to 2.77D (p<0.01), and the mean keratometry from 51.44 to 48.74D (p<0.02). As successful predictive factors, we found the asymmetry of keratoconus (p<0.01) in relation to the steepest meridian and patients implanted with a single ring (p<0.02). CONCLUSION: Defining preoperative successful predictive factors for Ferrara intracorneal ring segments is essential to enhance postoperative outcomes. PMID- 21632149 TI - [Quality of life of visually impaired adults after low-vision intervention: a pilot study]. AB - OBJECTIVE: To demonstrate the benefits of a low-vision intervention upon the quality of life of visually disabled adults. METHODS: The survey was proposed to patients who sought a low-vision intervention at the Colmar and Strasbourg hospital centres over a period of 9 months. Patients in agreement with the survey were asked to complete the 25-item National Eye Institute Visual Function Questionnaire (NEI VFQ25) in interview format by telephone, once they had attended the first meeting and again 2 months after the end of the low-vision intervention. RESULTS: The low-vision intervention led to overall improvement as judged by the 25 items of the questionnaire. Some items involving visual function and psychological issues showed significant benefits: the patients reported a more optimistic score concerning their general vision, described better nearby activities, and felt a bit more autonomous. CONCLUSION: More than mainstream psychological counselling, low-vision services help patients cope with visual disabilities during their daily life. The low-vision intervention improves physical and technical issues necessary to retaining autonomy in daily life. PMID- 21632150 TI - [Is myopia a risk factor for glaucoma?]. AB - Controversy exists in the literature concerning the role of axial myopia as a risk factor for primary open-angle glaucoma. Epidemiologic evidence suggests that moderate and especially high myopia with a refractive error exceeding -6D is a risk factor for the development and the progression of glaucomatous optic neuropathy, with a twofold to threefold increased risk of glaucoma compared with that of nonmyopic subjects. This risk has been proven to be independent of other glaucoma risk factors and intraocular pressure (IOP). Myopic eyes have slightly although probably not clinically relevant, higher IOPs than emmetropic or hyperopic eyes. Selection bias could account for some of the reported association between glaucoma and myopia given that myopic subjects are likely to consult their ophthalmologist more frequently and glaucoma is underdiagnosed in myopic patients due to the great variability of their optic disc morphology, especially in high myopia, and the difficulty to interpret their visual field. The weakness of the fibroglial matrix of the nerve fibers at the optic disc together with the structural alterations in the lamina cribrosa and choroid, could contribute to the high susceptibility of the optic disc to IOP fluctuations and to increasing the risk of glaucomatous neuropathy, especially in high myopic eyes. Special attention will be given to patients with mild myopia who present with both elevated IOP levels and a positive family history. On the other hand, high myopic subjects should be screened for glaucoma at closer intervals. Moreover, after appropriate adjustments for deviations in central corneal thickness have been made, IOP greater than 17 mmHg must already be regarded as critical and initiation of medical treatment considered. PMID- 21632151 TI - [Wolfram syndrome: clinical and genetic analysis in two sisters]. AB - Wolfram syndrome is a severe genetic disorder defined by the association of diabetes mellitus, optic atrophy, deafness, and diabetes insipidus. Two sisters complained of progressive visual loss. Fundus examination evidenced optic atrophy. Their past medical history revealed diabetes mellitus and deafness since childhood. The association of these symptoms made the diagnosis of Wolfram syndrome possible. It was confirmed by molecular analysis, which evidenced composite WFS1 heterozygous mutations inherited from both their mother and father. Ophthalmologists should be aware of the possibility of Wolfram syndrome when diagnosing optic atrophy in diabetic children. PMID- 21632152 TI - Infantile acropustulosis in internationally adopted children. AB - BACKGROUND: Infantile acropustulosis (IA) is a recurrent, self-limited, vesicopustular disorder affecting young children. Most cases occur after scabies infestation. IA seems to be common in children adopted from orphanages overseas. OBJECTIVES: We sought to demonstrate the prevalence of IA in internationally adopted children and to examine the number of doctors seen for IA before a diagnosis, the frequency of misdiagnoses, specialists most likely to make the diagnosis of IA, and features of IA. METHODS: An Internet-based survey was posted on international adoption forums. Parent participation was voluntary, and specific inclusion criteria existed. Follow-up telephone questionnaire was then conducted. RESULTS: Seventeen children had been given a diagnosis of IA and 21 had classic presentations but no IA diagnosis. Birth countries included Vietnam, China, Ethiopia, Guatemala, and Russia. Pediatric dermatologists and pediatricians affiliated with international adoption clinics were most likely to diagnose IA; 53% of diagnoses occurred after patient prompting. Frequent misdiagnoses were recurrent scabies and hand-foot-mouth disease. Feet were affected in 100% of cases and hands in 94%. Over 50% of children in both the diagnosed and undiagnosed groups had coexistent atopic dermatitis. LIMITATIONS: Limitations are potential parent reporting bias, selection bias, recall bias, and low response rate. Electronic survey instrument requires technically savvy parents. CONCLUSIONS: IA appears to be common in internationally adopted children, who spent early childhood in crowded, unclean living conditions with a high prevalence of scabies infestation. IA is frequently misdiagnosed as recurrent scabies, resulting in unnecessary use of permethrin. This study demonstrated a coexistence of atopic dermatitis in over 50% of IA cases. PMID- 21632153 TI - Effective monitoring of isotretinoin safety in a pediatric dermatology population: a novel "patient symptom survey" approach. AB - BACKGROUND: Assessment of adverse effects in pediatric patients on oral isotretinoin has not been standardized and the exact incidence is unknown. OBJECTIVE: Our goal was to determine the usefulness of an isotretinoin symptom survey as a screening tool for assessment and quantification of adverse effects, including psychiatric symptoms, during isotretinoin treatment in a pediatric population of different age groups. METHODS: We performed a retrospective chart review on a random sample of patients treated with isotretinoin at a tertiary pediatric dermatology clinic where patients completed an isotretinoin symptom survey at each visit. Responses were stratified by age group and psychiatric history. RESULTS: The charts of 102 patients, representing 123 courses of isotretinoin and 760 treatment-months, were reviewed. A total of 722 (95.0%) symptom surveys were complete and 38 (5.0%) were incomplete/missing. Recorded side effects were similar to published adult data; dry lips/dry skin were reported in 94.25% and 72.13% of treatment-months of isotretinoin, respectively. Psychiatric symptoms were reported in 1.65%, with no statistical difference between patients with or without a mental health history. Patients aged 11 to 15 years had similar side-effect profiles to those aged 16 to 21 years. Impaired night vision, nosebleeds, and dry/bloodshot eyes were more common in the older age group. LIMITATIONS: This was a retrospective chart review, with known limitations. The study was performed at a tertiary referral center for pediatric dermatology, possibly allowing patient selection bias. CONCLUSIONS: The isotretinoin symptom survey appears to be an effective screening tool to standardize monitoring of isotretinoin side effects in the pediatric population. PMID- 21632154 TI - [Training intervention studies with mini-tests to improve emergency medical care in a health center]. AB - OBJECTIVE: Test effectiveness and acceptability of interventions short essay-type training in health emergency management (EM). DESIGN: Combined case series and controlled study before and after training sessions. LOCATION: Health Center (HC). PARTICIPANTS: Team on duty, two monitors-facilitators, and a mannequin. MAIN MEASURES: VARIABLES: response times, staff performance, resource usage and opinion. Structure: scenarios and key messages. Instrument development: 1. Initial/final questionnaire and events. 2. Essential/non-essential times; 3. Post test opinion questionnaire. Performance of six consecutive 15 min tests fortnightly (including corrections) and poll after each test. A month later, repeat in random order and under similar conditions. ANALYSIS: repeated measures. RESULTS: A total of 93 (2/3) workers completed the initial survey, and 74 the final, with 46 participants (25 doctors, 7 nurses, 21 non-health completed 95 direct interventions. Matching participants > 80% between series. A reduction was seen in the "detection of collapse to first defibrillation" interval (10 to 4 min). EM events improved 2-3 fold and "sense of security during a real EM" increased from 23% to 71% among participants. The vast majority of participants said "useful corrections made by the facilitator." The proportions of those who "would like to see tests introduced" and those who said "re-training was needed in EM" were moderately increased (67.4% vs 85% in health care workers). The "would like to attempt basic life support" was unchanged. CONCLUSION: Despite being reduced in number and duration, this model of intervention has shown positive trends in terms of use and acceptability for implementation in the HC. PMID- 21632155 TI - Synthesis of novel alkyltriazole tagged pyrido[2,3-d]pyrimidine derivatives and their anticancer activity. AB - A series of novel alkyltriazole tagged pyrido[2,3-d]pyrimidine derivatives 5 and 6 was prepared starting from 2,3-active functional pyridine 1via cyclization, propargylation followed by reaction with alkyl or perfluoroalkyl azides under Sharpless conditions. All the compounds 5 and 6 were screened for anticancer activity against three cancer cell lines such as U(937), THP-1 and Colo205. The promising compounds 5b and 5e have been identified. PMID- 21632156 TI - In-vitro metabolic inhibition and antifertility effect facilitated by membrane alteration: search for novel antifertility agent using nifedipine analogues. AB - In search of non-hormonal male contraceptives, analogues of nifedipine, which causes reversible infertility, have been synthesized and their interaction at molecular level with model membrane has been probed. Analogues act differently with respect to their antifertility action. This is achieved by altering the cell metabolism thereby directly affecting the motility which is responsible for fertility. Secondly, these drugs bind differently to the interior of the cell membrane affecting the membrane fluidity, architecture and dynamics. Sulfasalazine and D(4) interact to a larger extent and alter the lipid bilayer phase to a hexagonal. D(1), D(2) and D(3) do not have considerable effect. D(4) is the most promising candidate as a lead compound for the development of novel non-hormonal male antifertility agents. PMID- 21632157 TI - Anti-inflammatory and anti-angiogenic effect of long chain n-3 polyunsaturated fatty acids in intestinal microvascular endothelium. AB - BACKGROUND & AIMS: The role of endothelial cells in inflammatory bowel disease has been recently emphasized. Endothelial activation and expression of adhesion molecules are critical for leukocytes recruitment into the inflammatory wall. Compelling evidence demonstrated anti-inflammatory effects of long chain n-3 PUFA in inflammatory models. We previously showed that long chain n-3 PUFA (EPA and DHA) inhibited inflammatory response in epithelial and dendritic cells. As long chain n-3 PUFA treatment led to a decreased expression of adhesion molecules in endothelial cells from other organs, we have now investigated their effect on intestinal endothelial cells in vitro and in colitic rats. METHODS: In vitro study: Primary culture of human intestinal microvascular endothelial cells (HIMEC) were pre-treated with DHA and then incubated with IL-1beta. In vivo study: Colitis was induced in 2 groups at day0 by intrarectal injection of 2-4-6 trinitrobenzen sulfonic acid (TNBS). Rats received by gavage either fish oil, rich in EPA and DHA (TNBS+n-3) or an isocaloric isolipidic oil formula for 14 days. RESULTS: DHA led to a decreased VCAM-1, TLR4, cyclooxygenase-2 and VEGFR2 expression and a decreased production of IL-6, IL-8 and GM-CSF and a reduced production of PGE(2) and LTB(4) (p < 0.001) in IL-1beta-induced HIMEC. Similarly, dietary intervention with fish oil rich in EPA and DHA significantly decreased colon production of PGE(2) and LTB(4,) endothelial VCAM-1 and VEGFR2 in rats with colitis. CONCLUSIONS: Data obtained from in vitro and in vivo studies reveal a potential anti-angiogenic role of long chain n-3 PUFA in intestinal endothelial cells. This protective effect of long chain n-3 PUFA may partly explain the observed benefit of dietary intake of long chain n-3 PUFA in IBD development. PMID- 21632158 TI - The effectiveness of a perinatal education programme on smoking, infant care, and psychosocial health for ethnic Turkish women. AB - OBJECTIVE: antenatal programmes might be effective in preventing unhealthy lifestyles, poor maternal infant care practices, and poor psychosocial health in ethnic minority women, but there are few evidence-based interventions. For this reason an antenatal education programme, called 'Happy Mothers, Happy Babies' (HMHB) was systematically designed for ethnic Turkish women in the Netherlands. DESIGN: in a non-randomised trial Turkish women attending HMHB (HMHB group) were compared with those receiving care as usual (control group). SETTING: Parent Child Centres, which provide integrated maternity and infant care. PARTICIPANTS: in both the HMHB (n=119) and the control (n=120) group, questionnaires were administered by ethnic Turkish interviewers at three (T0) and eight (T1) months of pregnancy, and two (T2) and six (T3) months after birth. FINDINGS: at baseline, women in the HMHB group had significantly lower educational levels, were less frequently in paid employment, had less knowledge about smoking, and showed more often mildly depressive symptoms. Adjusted analyses showed that HMHB was effective in improving knowledge about smoking (OR=2.73; 95% CI 1.40, 5.31), intention to engage in prevention of sudden infant death syndrome (SIDS) (OR=8.08; 95% CI 3.34, 19.56) and short-term SIDS prevention behaviour (OR=2.22; 95% CI=1.18, 4.19). However, no intervention effect was found for smoking during pregnancy, SIDS prevention behaviour on the long term, soothing behaviour, serious depressive symptoms, and parent-child attachment. KEY CONCLUSIONS: although we could not demonstrate intervention effects on all outcome measures, the HMHB programme appears to be highly welcome, and reaches an underserved minority group at increased risk for adverse perinatal outcomes. IMPLICATIONS FOR PRACTICE: the HMHB programme is one of the first systematically developed antenatal interventions for ethnic minority women. The programme can be used as a basic antenatal programme, and as a screening opportunity for women who smoke or show serious depressive symptoms. PMID- 21632159 TI - Relationships between clinical measures of static foot posture and plantar pressure during static standing and walking. AB - BACKGROUND: Information is limited about the relationships between clinical measures of static foot posture and peak plantar pressures under the medial column of the foot. The purpose was to examine these relationships during static standing and walking. METHODS: A single-group exploratory design using correlation and regression was used to determine relationships. Ninety-two healthy volunteers participated. Clinical measures of static foot posture including arch index, navicular drop and navicular drift were obtained during static standing. Peak plantar pressures under the hallux, medial forefoot, medial midfoot, and medial rearfoot were obtained during standing and walking. FINDINGS: Static foot posture was related to peak plantar pressures during standing and walking, but the strengths of relationships ranged from poor to fair. During standing, navicular drop was correlated (P<=0.05) with hallux (r=0.29) and medial forefoot (r=-0.17) pressures, while arch index (r=-0.17) and navicular drift (r=0.25) were correlated (P<=0.05) with hallux pressure. During walking, arch index, navicular drop and navicular drift were correlated (P<=0.05) with hallux and medial forefoot pressures (r range -0.30 to 0.41), while arch index (r=-0.15) and navicular drop (r=0.16) were correlated (P<=0.05) with medial midfoot pressure. Regression models predicted (P<=0.05) hallux (R(2)=0.08) and medial midfoot (R(2)=0.05) pressures during standing, and hallux (R(2)=0.18), medial forefoot (R(2)=0.07), and medial rearfoot (R(2)=0.05) pressures during walking. INTERPRETATION: In healthy participants, lower arch foot postures are associated with greater pressures under the hallux and medial mid-foot and lower pressures under the medial forefoot, but the strength of these relationships may be only poor to fair. PMID- 21632160 TI - When does neighbourhood matter? Multilevel relationships between neighbourhood social fragmentation and mental health. AB - Studies investigating relationships between mental health and residential areas suggest that certain characteristics of neighbourhood environments matter. After developing a conceptual model of neighbourhood social fragmentation and health we examine this relationship (using the New Zealand Index of Neighbourhood Social Fragmentation (NeighFrag)) with self-reported mental health (using SF-36). We used the nationally representative 2002/3 New Zealand Health Survey dataset of urban adults, employing multilevel methods. Results suggest that increasing neighbourhood-level social fragmentation is associated with poorer mental health, when simultaneously accounting for individual-level confounding factors and neighbourhood-level deprivation. The association was modified by sex (stronger association seen for women) and labour force status (unemployed women more sensitive to NeighFrag than those employed or not in labour force). There was limited evidence of any association of fragmentation with non-mental health outcomes, suggesting specificity for mental health. Social fragmentation as a property of neighbourhoods appears to have a specific association with mental health among women, and particularly unemployed women, in our study. PMID- 21632161 TI - Young adults' decision making surrounding heavy drinking: a multi-staged model of planned behaviour. AB - This paper examines the real life contexts in which decisions surrounding heavy drinking are made by young adults (that is, on occasions when five or more alcoholic drinks are consumed within a few hours). It presents a conceptual model that views such decision making as a multi-faceted and multi-staged process. The mixed method study draws on purposive data gathered through direct observation of eight social networks consisting of 81 young adults aged between 18 and 25 years in Perth, Western Australia, including in-depth interviews with 31 participants. Qualitative and some basic quantitative data were gathered using participant observation and in-depth interviews undertaken over an eighteen month period. Participants explained their decision to engage in heavy drinking as based on a variety of factors. These elements relate to socio-cultural norms and expectancies that are best explained by the theory of planned behaviour. A framework is proposed that characterises heavy drinking as taking place in a multi-staged manner, with young adults having: 1. A generalised orientation to the value of heavy drinking shaped by wider influences and norms; 2. A short-term orientation shaped by situational factors that determines drinking intentions for specific events; and 3. An evaluative orientation shaped by moderating factors. The value of qualitative studies of decision making in real life contexts is advanced to complement the mostly quantitative research that dominates research on alcohol decision making. PMID- 21632162 TI - Representational momentum in older adults. AB - Humans have a tendency to perceive motion even in static images that simply "imply" movement. This tendency is so strong that our memory for actions depicted in static images is distorted in the direction of implied motion - a phenomenon known as representational momentum (RM). In the present study, we created an RM display depicting a pattern of implied (clockwise) rotation of a rectangle. Young adults viewers' memory of the final position of the test rectangle was biased in the direction of continuing rotation, but older adults did not show a similar memory bias. We discuss several possible explanations for this group difference, but argue that the failure of older adults to shown an RM effect most likely reflects age-related changes in areas of the brain involved in processing real and implied motion. PMID- 21632163 TI - [Gender and health status response to long-term oxygen therapy in COPD patients]. AB - INTRODUCTION: The association of gender with health status (HS) response to long term oxygen therapy (LTOT) in very severe COPD is unclear. The aims of this study were: (1) to compare dyspnea perception and HS between male and female with very severe COPD at baseline and (2) to provide a prospective assessment of HS response to LTOT, according to gender. PATIENTS AND METHODS: Hypoxemic COPD (n = 97, age: 65.5 +/- 9.6 years, 53% males) were enrolled in a prospective longitudinal study over 12 months or until death. St. George's Respiratory Questionnaire (SGRQ) and baseline dyspnea index (BDI) were assessed. RESULTS: At baseline, HS impairment and dyspnea sensation were similar between genders. After 12 months of LTOT, women presented improvement in symptom (64.1 +/- 20.6 versus 40.6 +/- 22.9; P < 0.0001) and total SGRQ scores. Men also showed improvement in symptoms after 12 months (62.7 +/- 23.3 versus 49.6 +/- 22.8; P < 0.0005); however, they presented deterioration of activity, impact and total scores during the study period, with markedly decline of activity domain (68.5 +/- 20.0 versus 75.9 +/- 16.9; P = 0.008). BDI did not show significant difference by gender over the study period. CONCLUSIONS: Our results show that the HS course in very severe COPD patients differs according to gender, as females show greater response longitudinally to LTOT. PMID- 21632164 TI - [Mitral valve repair: echocardiography is its best friend]. PMID- 21632165 TI - Hand position for chest compression. PMID- 21632166 TI - Value of post-resuscitation electrocardiogram in the diagnosis of acute myocardial infarction in out-of-hospital cardiac arrest patients. AB - BACKGROUND: Diagnosis of acute myocardial infarction (AMI) in out-of-hospital cardiac arrest (OHCA) patients is important because immediate coronary angiography with coronary angioplasty could improve outcome in this setting. However, the value of acute post-resuscitation electrocardiographic (ECG) data for the detection of AMI is debatable. METHODS: We assessed the diagnostic characteristics of post-resuscitation ECG changes in a retrospective single centre study evaluating several ECG criteria of selection of patients undergoing AMI, in order to improve sensitivity, even at the expense of specificity. Immediate post resuscitation coronary angiogram was performed in all patients. AMI was defined angiographically using coronary flow and plaque morphology criteria. RESULTS: We included 165 consecutive patients aged 56 (IQR 48-67) with sustained return of spontaneous circulation after OHCA between 2002 and 2008. 84 patients had shockable, 73 non-shockable and 8 unknown initial rhythm; 36% of the patients had an AMI. ST-segment elevation predicted AMI with 88% sensitivity and 84% specificity. The criterion including ST-segment elevation and/or depression had 95% sensitivity and 62% specificity. The combined criterion including ST segment elevation and/or depression, and/or non-specific wide QRS complex and/or left bundle branch block provided a sensitivity and negative predictive value of 100%, a specificity of 46% and a positive predictive value of 52%. CONCLUSION: In patients with OHCA without obvious non-cardiac causes, selection for coronary angiogram based on the combined criterion would detect all AMI and avoid the performance of the procedure in 30% of the patients, in whom coronary angiogram did not have a therapeutic role. PMID- 21632167 TI - A sternal accelerometer does not impair hemodynamics during piglet CPR. AB - AIM: To determine whether the residual weight of a 260 g sternal accelerometer/force feedback device (AFFD) adversely affects hemodynamics during cardiopulmonary resuscitation in a piglet model of ventricular fibrillation cardiac arrest. METHODS: After induction of ventricular fibrillation, cardiopulmonary resuscitation was provided to ten piglets (10.8 +/- 1.9 kg) for 12 min while maintaining aortic systolic pressure of 80-90 mm Hg during four 3 min periods with or without an AFFD on the chest. Cardiac output and left ventricular myocardial blood flow were determined by neutron-microsphere technique. RESULTS: Using a linear mixed-effect model with residual maximum likelihood estimation to control for changes in cardiopulmonary resuscitation hemodynamics over time, cardiac output and myocardial blood flow did not differ with AFFD versus without AFFD. During the first 6 min, mean (+/- SEM) cardiac outputs were 0.42 (+/- 0.05)L/min with AFFD versus 0.31 (+/- 0.04)L/min without AFFD, and median left ventricular myocardial blood flows were 40.5 (+/- 7.3)mL/min/100g with AFFD versus 40.4 (+/- 5.0)mL/min/100g without AFFD. The mean right atrial diastolic pressures and coronary perfusion pressures were also not different (8 +/- 0.7 mm Hg versus 8 +/- 0.9 mm Hg and 16 +/- 2 mm Hg versus 16 +/ 2 mm Hg, respectively, during the first 6 min of CPR). CONCLUSION: The use of a 260 g accelerometer/force feedback device designed for real-time feedback to the rescuer during resuscitation efforts did not adversely affect cardiac output or left ventricular myocardial blood flow during 12 min of chest compressions in a piglet model of ventricular fibrillation cardiac arrest. PMID- 21632168 TI - Systolic blood pressure below 110 mm Hg is associated with increased mortality in blunt major trauma patients: multicentre cohort study. AB - INTRODUCTION: Non-invasive systolic blood pressure (SBP) measurement is often used in triaging trauma patients. Traditionally, SBP< 90 mm Hg has represented the threshold for hypotension, but recent studies have suggested redefining hypotension as SBP < 110 mm Hg. This study aims to examine the association of SBP with mortality in blunt trauma patients. METHODS: This is an analysis of prospectively recorded data from adult (>= 16 years) blunt trauma patients. Included patients presented to hospitals belonging to the Trauma Audit and Research Network (TARN) between 2000 and 2009. The primary outcome was the association of SBP and mortality rates at 30 days. Multivariate logistic regression models were used to adjust for the influence of age, gender, Injury Severity Score (ISS) and Glasgow Coma Score (GCS) on mortality. RESULTS: 47,927 eligible patients presented to TARN hospitals during the study period. Sample demographics were: median age: 51.1 years (IQR=32.8-67.4); male 60% (n=28,694); median ISS 9 (IQR=8-10); median GCS 15 (IQR=15-15); and median SBP 135 mm Hg (IQR=120-152). We identified SBP< 110 mm Hg as a cut off for hypotension, where a significant increase in mortality was observed. Mortality rates doubled at < 100 mm Hg, tripled at < 90 mm Hg and were 5- to 6-fold at < 70 mm Hg, irrespective of age. CONCLUSION: We recommend triaging adult blunt trauma patients with a SBP< 110 mm Hg to resuscitation areas within dedicated trauma units for close monitoring and appropriate management. PMID- 21632169 TI - Evaluation and management of acute menorrhagia in women with and without underlying bleeding disorders: consensus from an international expert panel. AB - Acute menorrhagia is a common gynecological disorder. Prevalence is high among women with inherited bleeding disorders and recent guidance for optimal management is lacking. Following a comprehensive review of the literature, an international expert panel in obstetrics, gynecology and hematology reached consensus on recommendations regarding the management of acute menorrhagia in women without a diagnosed bleeding disorder, as well as in patients with von Willebrand disease, platelet function disorders and other rare hemostatic disorders. The causes and predictors of acute menorrhagia are discussed and special consideration is given for the treatment of women on anticoagulation therapy. This review and accompanying recommendations will provide guidance for healthcare practitioners in the emergency management of acute menorrhagia. PMID- 21632170 TI - Current status of surrogacy in Japan and uterine transplantation research. AB - Recent advances in assisted reproductive technology (ART) have made it possible to circumvent many causes of male and female infertility. The right to have a child by ART has been respected for infertile couples. However, there are currently no legal regulations concerning ART in Japan, and this has resulted in social and ethical problems. Surrogacy involves particularly complex medical, ethical, social, and legal issues, and is frequently focused on as a major social concern. Uterine transplantation (UTx) is a potential alternative for young women with uterine factor infertility due to hysterectomy for treatment of a malignant uterine tumor or massive blood loss after delivery, or because of a congenital disease such as Mayer-Rokitansky-Kuster syndrome. UTx has been examined in experimental animals as a basis for establishment of fecundity for young women with uterine factor infertility. In this review, we focus on surrogacy in Japan and UTx research, and discuss the current status and concerns in this field. PMID- 21632171 TI - An overview of biomarkers for the ovarian cancer diagnosis. AB - Even though there are a lot of options in treating gynecological malignancies, ovarian cancer still remains a leading cause of death. Diagnosis at an early stage is the most important determinant of survival. Current diagnostic tools applied at clinics have had very limited success in early detection. Discovery of new diagnostic biomarkers/panels for early diagnosis of ovarian cancer is one of the main challenges of modern medicine. With the progress of techniques in genomics and proteomics, numerous molecular biomarkers/panels were identified and showed promise for ovarian cancer diagnosis, but still need further validation. This article summarizes various types of markers investigated by different strategies/technologies for the ovarian cancer diagnosis at present, including gene-, protein-based and emerging ovarian cancer indicators (such as microRNA-, metabolite-based). Before biomarker tests are translated for routine use, more researches, such as retrospective and prospective clinical trials, are needed to evaluate the overall clinical utility of the tests. PMID- 21632172 TI - Critical care and transfusion management in maternal deaths from postpartum haemorrhage. AB - OBJECTIVES: In postpartum haemorrhage (PPH), as for other causes of acute haemorrhage, management can have a major impact on patient outcomes. The aim of this study was to describe critical care management, particularly transfusion practices, in cases of maternal deaths from PPH. STUDY DESIGN: This retrospective study provided a descriptive analysis of all cases of maternal death from PPH in France identified through the systematic French Confidential Enquiry into Maternal Death in 2000-2003. RESULTS: Thirty-eight cases of maternal death from PPH were analysed. Twenty-six women (68%) had a caesarean section [21 (55%) emergency, five (13%) elective]. Uterine atony was the most common cause of PPH (n=13, 34%). Women received a median of 9 (range 2-64) units of red blood cells (RBCs) and 9 (range 2-67) units of fresh frozen plasma (FFP). The median delay in starting blood transfusion was 82 (range 0-320)min. RBC and FFP transfusions peaked 2-4h and 12-24h after PPH diagnosis, respectively. The median FFP:RBC ratio was 0.6 (range 0-2). Fibrinogen concentrates and platelets were administered to 18 (47%) and 16 (42%) women, respectively. Three women received no blood products. Coagulation tests were performed in 20 women. The haemoglobin concentration was only measured once in seven of the 22 women who survived for more than 6h. Twenty-four women received vasopressors, a central venous access was placed in 11 women, and an invasive blood pressure device was placed in two women. General anaesthesia was administered in 37 cases, with five patients being extubated during active PPH. CONCLUSIONS: This descriptive analysis of maternal deaths from PPH suggests that there may be room for improvement of specific aspects of critical care management, including: transfusion procedures, especially administration delays and FFP:RBC ratio; repeated laboratory assessments of haemostasis and haemoglobin concentration; invasive haemodynamic monitoring; and protocols for general anaesthesia. PMID- 21632173 TI - EAU guidelines on testicular cancer: 2011 update. AB - CONTEXT: On behalf of the European Association of Urology (EAU), guidelines for the diagnosis, therapy, and follow-up of testicular cancer were established. OBJECTIVE: This article is a short version of the EAU testicular cancer guidelines and summarises the main conclusions from the guidelines on the management of testicular cancer. EVIDENCE ACQUISITION: Guidelines were compiled by a multidisciplinary guidelines working group. A systematic review was carried out using Medline and Embase, also taking Cochrane evidence and data from the European Germ Cell Cancer Consensus Group into consideration. A panel of experts weighted the references, and a level of evidence and grade of recommendation were assigned. RESULTS: There is a paucity of literature especially regarding longer term follow-up, and results from a number of ongoing trials are awaited. The choice of treatment centre is of the utmost importance, and treatment in reference centres within clinical trials, especially for poor-prognosis nonseminomatous germ cell tumours, provides better outcomes. For patients with clinical stage I seminoma, based on recently published data on long-term toxicity, adjuvant radiotherapy is no longer recommended as first-line adjuvant treatment. The TNM classification 2009 is recommended. CONCLUSIONS: These guidelines contain information for the standardised management of patients with testicular cancer based on the latest scientific insights. Cure rates are generally excellent, but because testicular cancer mainly affects men in their third or fourth decade of life, treatment effects on fertility require careful counselling of patients, and treatment must be tailored taking individual circumstances and patient preferences into account. PMID- 21632174 TI - Analytical solutions for exposures and toxic loads in well-mixed shelters in support of shelter-in-place assessments. AB - Understanding the exposure and toxic load for the interior of buildings during and following the passage of an external airborne hazard can be a critical piece of information in deciding the benefit from adopting a shelter-in-place strategy. Whilst numerical methods allow the calculation of such parameters for the general case, analytical solutions allow more rapid assessments to be made and highlight the key parameters more clearly. Analytical expressions are derived for the exposure due to the acute inhalation of toxic chemicals and the associated toxic load as a function of time, external hazard duration and building air change rate assuming a top-hat outdoor concentration profile and no indoor loss mechanism. It is shown that the internal exposure tends to the external exposure at long times for any external concentration profile. Expressions are derived for toxic loads with exponents n=m/2 where 2 <= m <= 7 is an integer to cover the range of typical values (1 <= n <= 3.5). At long times the ratio of internal to external toxic load for a top-hat outdoor concentration profile is shown to be a function of the product of the air change rate and the duration of the external hazard. PMID- 21632175 TI - Aqueous two-phase systems: a new approach for the determination of p-aminophenol. AB - A new method has been developed for the spectrophotometric determination of p aminophenol (PAP) in water, paracetamol formulations and human urine samples with a recovery rate between 94.9 and 101%. This method exploits an aqueous two-phase system (ATPS) liquid-liquid extraction technique with the reaction of PAP, sodium nitroprusside and hydroxylamine hydrochloride in pH 12.0, which produces the [Fe(2)(CN)(10)](10-) anion complex that spontaneously concentrates in the top phase of the ATPS ([Formula in text]). The ATPS does not require an organic solvent, which is a safer and cleaner liquid-liquid extraction technique for the determination of PAP. The linear range of detection was from 5.00 to 500 MUg kg( 1) (R >= 0.9990; n=8) with a coefficient of variation of 2.11% (n=5). The method exhibited a detection limit of 2.40 MUg kg(-1) and a quantification limit of 8.00 MUg kg(-1). The ATPS method showed a recovery that ranged between 96.4 and 103% for the determination of PAP in natural water and wastewater samples, which was in excellent agreement with the results of the standard 4-aminoantipyrine method that was performed on the same samples. PMID- 21632176 TI - Using response surface methodology to assess the effects of iron and spent mushroom substrate on arsenic phytotoxicity in lettuce (Lactuca sativa L.). AB - The effects of iron (Fe) and spent mushroom substrate (SMS) arsenic (As) phytotoxicity towards lettuce in artificial soils were investigated to separate the adverse soil parameters relating to As toxicity using a response surface methodology. SMS induced the root elongation of lettuce in both control and As treated soils. However, in phytotoxicity test using a median effective concentration (EC(50)) of As, Fe and the interaction between both parameters (Fe*SMS) significantly affected EC(50), which explained 71% and 23% of the response, respectively. The refined model was as follows: EC(50) of As (mgkg( 1))=10.99+60.03 * Fe-10.50 * Fe*SMS. The results confirmed that the soil parameters relating to the As mobility in soils were important factors affecting its toxicity. In conclusion, Fe significantly reduced the As phytotoxicity. However, although SMS enhanced the root elongation, SMS in As-treated soils decreased EC(50) of As on the root growth via its interaction with Fe. Despite the limitations of the artificial soils and range of parameters studied, the application of this statistical tool can be considered a powerful and efficient technique for interpretation and prediction of the complicated results caused by the interactions between many factors within the soil environments. PMID- 21632177 TI - A new model of repulsive force in eddy current separation for recovering waste toner cartridges. AB - Eddy current separation (ECS) is an efficient method for separating aluminum from plastic in crushed waste toner cartridge (TCs). However, in China, ECS quality of aluminum from plastic is rather low in production practice. Repeating separation even manual sorting is required in the production. Improving separation quality of aluminum has been the pressing problem in the recovery of waste TCs. Furthermore, improving ECS quality can reduce the secondary-pollution (furan and dioxin) brought by plastic in later smelting process for the purification of recovered aluminum. Thus, a new model of repulsive force containing impact factors (machine: B(r), k, R, S(m), B(m); material: S(p), V, gamma; and operation: omega(m), v, delta) of the separation process was constructed for guiding the ECS process of waste TCs recovering in this paper. For testing whether the model of repulsive force was suitable to guide the ECS, calculation and experiment of detachment angle of aluminum flake were studied. The calculation results of the detachment angles were agreed with the testing experiment. It indicates that the model is suitable for guiding the ECS of waste TCs recovering. PMID- 21632178 TI - NO treated TiO2 as an efficient visible light photocatalyst for NO removal. AB - In this study, we report that nitrogen doped TiO(2) could be achieved via thermal treatment of Degussa P25 TiO(2) in NO atmosphere directly (P25-NO). The samples were characterized with XRD, XPS, and FT-IR. The characterization results suggested that nitrogen species were interstitially doped in P25-NO during the NO thermal treatment process. In comparison with P25, the P25-NO exhibited significantly enhanced photocatalytic activities under visible light irradiation (lambda>420 nm) for gaseous NO removal. On the basis of electronic band structure theory, we proposed a possible mechanism for the enhanced visible light driven photocatalytic oxidation process over the interstitial N doping P25-NO samples. This work could not only deepen understanding of the enhanced photoactivity originated from interstitial N doping in TiO(2), but also provide a facile route to prepare nitrogen doped TiO(2) for environmental and energy applications. PMID- 21632179 TI - Emulsion polymerization of vinyl acetate: safe optimization of a hazardous complex process. AB - Fast and exothermic discontinuous emulsion polymerization processes are particularly difficult to optimize from both safety and productivity point of view because of the occurrence of side undesired reactions (e.g. chain transfer to monomer, backbiting, propagation of tertiary radicals, termination by disproportion, etc.) and the hazards of boiling phenomena and stable foam formation under atmospheric pressure. Moreover, the relevant number of loading, heating and cooling steps, required before starting the monomer addition (that is, the desired reaction), makes a strict product quality reproducibility very difficult to obtain. Under these operating conditions, it is necessary to employ a suitable combined theoretical and experimental procedure able to detect the optimum process dosing time at both the laboratory and the industrial scale. In this work, it is shown how to use the topological criterion theory together with proper adiabatic calorimeter and RC1 experimental data to safely optimize the synthesis of polyvinyl acetate through the radical emulsion polymerization of vinyl acetate by the means of an indirectly cooled isoperibolic semibatch reactor. PMID- 21632180 TI - High pressure processing treatment prevents embryonation of eggs of Trichuris vulpis and Ascaris suum and induces delay in development of eggs. AB - High hydrostatic pressure processing (HPP) is an effective non-thermal treatment used to inactivate pathogens from a variety of food and food products. It has been extensively examined using prokaryotic organisms and protozoan's but has had limited study on metazoans. Treatment using HPP has been shown to be effective in inactivating nematode larvae in food and preventing embryonation of Ascaris suum eggs. We conducted experiments using eggs of the canine whipworm Trichuris vulpis collected from naturally infected dogs and A. suum eggs from naturally infected pigs. We observed a delay in development of eggs of T. vulpis in a preliminary experiment and conducted 2 experiments to test the hypothesis that appropriate HPP levels can induce a delay in embryonation of nematode eggs. In experiment 1, nonembryonated T. vulpis eggs in tap water were packaged in sealable bags and exposed to 138-600 megapascals (MPa; 1 MPa=10 atm=147 psi) for 60s in a commercial HPP unit. In a second experiment, nonembryonated eggs of A. suum were exposed to 138-600 MPa and treated for 60s in the same commercial HPP unit. Embyronation of T. vulpis eggs was delayed by 4 and 5 days for eggs treated with 207 and 241 MPa but eventually eggs developed and the numbers of embryonated eggs was similar to controls on day 55. Embryonation of T. vulpis eggs treated with 345 or 350 MPa was delayed by 9 days and never reached more than 5% of eggs embryonated. On day 55 post treatment, 95% of control nontreated T. vulpis eggs were embryonated, 100-65% of eggs treated with 138-276 MPa were embryonated, a maximum of 5% of eggs treated with 345-350 MPa were embryonated, and 0% of eggs treated with >= 400 MPa were embryonated. T. vulpis eggs treated with >= 400 MPa did not undergo cell division. Embryrnation of A. suum eggs was delayed by 4, 10, and 16 days for eggs treated with 207, 241, and 250MPa, respectively, compared to nontreated control eggs. A. suum eggs treated with 207 MPa eventually embryonated to similar % embryonation values as controls and 138 MPa treated eggs but eggs treated with 241 or 250 MPa were always <5% embryonated. A. suum eggs treated with >= 300 MPa did not undergo cell division. On the final day of examination at day 56 after treatment, the % of embryonated eggs was 92% nontreated controls, 94% treated with 138 MPa, 84% treated with 207 MPa, 2% treated with 241 or 250 MPa, and 0% treated with 276, 200, 345, 400, or 414 MPa, respectively. PMID- 21632181 TI - Immunogenicity and protective efficacy of two recombinant pseudorabies viruses expressing Toxoplasma gondii SAG1 and MIC3 proteins. AB - Toxoplasma gondii is one of the most common parasitic pathogens in humans and warm-blooded animals, causing toxoplasmosis. One of the efficient ways to control this disease is immunization. In this study, two recombinant pseudorabies virus (PRV) expressing TgSAG1 (rPRV-SAG1) and TgMIC3 (rPRV-MIC3) based on the PRV vaccine strain were developed by homologous recombination and used for immunizing BALB/c mice. Ninety BALB/c mice were randomly divided into five groups including four experimental groups (inoculated twice in 4 weeks interval with PRV TK-/gG /EGFP+, rPRV-SAG1, rPRV-MIC3, rPRV-SAG1+rPRV-MIC3, respectively) and one control group (inoculated with medium). All mice vaccinated with rPRV developed a high level of specific antibody responses against T. gondii lysate antigen (TLA), a strong increase of the splenocyte proliferative response, and significant levels of IFN-gamma and IL-2 production. These results demonstrated that rPRV could induce significant humoral and cellular Th1 immune responses. Moreover, rPRV immunization induced partial protection against a lethal challenge with T. gondii RH strain, and neutralizing antibodies against PRV in a BALB/c mouse model. The mice immunized with the rPRV-SAG1 and rPRV-MIC3 cocktail could develop higher T. gondii-specific IgG antibodies and lymphocyte proliferative responses and conferred more efficient protection against T. gondii challenge. These results suggested that expression of protective antigens of T. gondii in PRV is a novel approach towards the development of a vaccine against both animal pseudorabies and toxoplasmosis. PMID- 21632182 TI - The anticancer effects of Vitamin D and omega-3 PUFAs in combination via cod liver oil: one plus one may equal more than two. AB - In the past number of years, the anticancer activities of omega-3 polyunsaturated fatty acids (omega3-PUFAs) as well as Vitamin D have been intensively studied, but separately. Supplementation of Vitamin D and omega-3 PUFA via cod-liver oil, one of few natural sources of both of these molecules, may have additive and possibly synergistic anticancer effects. Cod-liver oil has been used effectively to treat diseases such as Rheumatism but has not been studied as an anticancer agent. This review examines the prominent, striking and possibly important similarities between the anticancer effects of omega3-PUFAs and Vitamin D metabolites as well as the possible overlapping signaling pathways by which they may operate. The mechanisms that will be examined in this review fall broadly under the categories of being anti-inflammatory, pro-apoptotic, anti-angiogenic and anti-proliferative. Finally, we compare the potential for use of omega3 PUFAs, Vitamin D combinatorial supplementation both in prevention and treatment of disease. Some data also suggests that the timing of supplementation modifies the effects of Vitamin D and omega3 fatty acids. PMID- 21632183 TI - Effect of an elevated monounsaturated fat diet on pork carcass and meat quality traits and tissue fatty acid composition from York-crossed barrows and gilts. AB - Fifty-seven York-crossed barrows and gilts were fed either a grain and soy diet (CONTROL with 28% C18:1) or a similar diet enriched with oleic acid (HO with 43% C18:1, Greedy-Grass OLIVA(r)). There were no interactions between dietary treatment and gender and there were no differences in intramuscular and subcutaneous fatty acid composition between sexes (P>0.05). Similar primal cut yields, composition of major primal cuts, and carcass and meat quality characteristics were found for HO and CONTROL fed pigs. Apart from a few significant but small differences for some fatty acids, intramuscular fatty acid composition was similar for both dietary treatments. Subcutaneous fat from HO fed pigs had a 6.9% increase in total monounsaturated fatty acids and a 9.3% reduction in total polyunsaturated fatty acids (P<0.05) compared with CONTROL fed animals, without adversely affecting carcass quality and producing suitable hams for processing by the meat industry. PMID- 21632184 TI - Possible biological markers of the time of processing of dry-cured ham. AB - Several muscle compounds (creatine, creatinine, hypoxanthine, inosine, inosine 5' monophosphate, xanthine, adenosine monophosphate, guanosine, and uridine) were studied as possible biological markers of a minimum dry-cured ham processing time. A correlation between the concentration of the compounds and the time of processing was found. The ratios for some of them were calculated to study their behaviour during processing. The Hx/Ino ratio substantially increased up to 5 months of curing and then remained constant (p<0.05). The Hx/Ino ratio might be considered as a potential biomarker of the minimum time of dry-cured processing (5 months). The Cn/Cr ratio increased during drying up to 9 months of ripening (p<0.05). However, although Cn/Cr ratios remained constant after 9 months of processing, variations between hams were observed due to the differences existing in the raw meats and small differences in processing conditions, making it difficult to consider Cn/Cr ratios as biomarkers of ripening time. PMID- 21632185 TI - Lessons to learn about postmortem metabolism using the AMPKgamma3(R200Q) mutation in the pig. AB - The development of pork quality attributes - including color, water holding capacity, and texture - are controlled largely by the rate and extent of postmortem pH decline. In turn, pH decline is thought to follow the anaerobic degradation of glycogen to "lactic acid". The "Hampshire effect" or Rendement Napole (RN(-)) pig, which has elevated muscle glycogen, propagated the widespread use of "glycolytic potential" to describe muscle's capacity for postmortem glycolysis. Since the evolution of glycolytic potential, there have been great strides in molecular and technical capabilities. Hence, it must be appreciated that the RN(-) pig possesses a mutation in the gene encoding the AMP-activated protein kinase (AMPK) gamma3 subunit, which results in a non-conservative amino acid substitution (R200Q). AMPK, a major energy sensor in skeletal muscle, influences enzyme activity, gene and protein expression, fiber type, and mitochondrial biogenesis. The utility of glycolytic potential as indicator of ultimate pH should be reevaluated in the context of the metabolic differences between AMPKgamma3 mutated and normal muscle. Understanding the metabolism and energetics in AMPKgamma3 mutated muscle may provide insight into the mechanisms influencing the rate and extent of postmortem metabolism. PMID- 21632186 TI - Correlation between E-cadherin and CD44 adhesion molecules expression and cervical lymph node metastasis in oral tongue SCC: Predictive significance or not. AB - The aim of this article was to evaluate the expression of E-cadherin and CD44 adhesion molecule in oral tongue squamous cell carcinoma (SCC) since inappropriate expression of adhesion molecules raises the metastatic ability of the tumor cells. Biopsy specimens from 92 patients with tongue SCC were examined for the expression of E-cadherin and CD44 by immunohistochemistry. The relationship of immunoreactivity with tumor stage and cervical lymph node metastasis was then analyzed. Sixty-one patients (66.3%) had reduced or negative staining for CD44. Weak or absent staining for E-cadherin was seen in 14 patients (15.21%). Cervical lymph node metastasis is associated with decreased or negative staining for CD44, but no association was found between E-cadherin immunoreactivity and nodal metastasis. Our study reveals that reduced expression of CD44 could be an indicator of high invasiveness of tumor by increasing cervical lymph node metastasis. PMID- 21632187 TI - The association between parental risk behaviors during childhood and having high risk networks in adulthood. AB - BACKGROUND: Prior research suggests that both social networks and parent drug use influence individual drug use among adolescents and that peers continue to influence drug use among adults. This analysis aims to determine whether parent drug use during childhood is associated with having drug-using networks in adulthood after adjusting for individual adult drug use. METHODS: 650 young adult drug users were recruited through targeted street outreach and respondent-driven sampling in New York City (2006-2009). Baseline surveys ascertained demographics, network characteristics, drug use behaviors, and parental drug use during childhood. Negative binomial regression was used to evaluate this association. RESULTS: The median age was 33 years, 22% injected, 49% were Black, and during childhood 26% of mothers, 32% of fathers, and 13% of primary caregivers used drugs. After adjustment, having >1 parent who used drugs was associated with having a greater proportion of drug using (Adjusted Prevalence Ratio [APR]=1.18; 95%CI: 1.01-1.38) and specifically crack-smoking networks (APR=1.71; 95%CI: 1.21 2.43) in adulthood. Females' networks consisted of more drug users (APR=1.18; 95%CI: 1.01-1.38), injectors (APR=1.44; 95%CI: 1.09-1.90), crack smokers (APR=1.48; 95%CI: 1.18-1.87) and heroin users (APR=1.43; 95%CI: 1.13-1.81); blacks had a greater proportion of crack smoking (APR=1.41; 95%CI: 1.09-1.82), but a smaller proportion of injecting (APR=0.64; 95%CI: 0.43-0.94) and heroin smoking (APR=0.60; 95%CI: 0.47-0.77) networks as adults. CONCLUSIONS: These data suggest that parental drug use is independently associated with having drug-using networks in adulthood. Interventions that target parents and caregivers and that promote drug cessation could impede risky network formation in both adolescents and adults. PMID- 21632188 TI - Pathways to treatment retention for individuals legally coerced to substance use treatment: the interaction of hope and treatment motivation. AB - BACKGROUND: Although several states have adopted policies diverting individuals convicted of non-violent drug offenses to substance use treatment, in lieu of incarceration or as a condition of probation, previous research has produced inconsistent findings on the effectiveness of such programs when comparing outcomes for legally coerced individuals to more voluntary entrants. Less studied in these populations is within group variation in treatment expectations and motivation influences, which have been shown to affect retention as well. METHODS: As motivation has traditionally been viewed as contributing to treatment retention and higher levels of hope (the perception that goals can be met) are viewed as an asset in treatment, the role of these factors in predicting better retention between legally coerced and more voluntary clients were examined in a sample of 289 treatment admissions in California. RESULTS: Results found that motivation mediates the relationship between hope and retention for participants in general. Although the differences in mediation between the legally coerced and the non-legally coerced were not significant, when examining the groups separately, there was a significant mediation of the relationship between hope and retention by motivation only for those individuals who were not legally coerced into treatment (p<.05). CONCLUSION: The findings imply that while being legally coerced may lead to different pathways to treatment retention, for individuals who were not legally coerced, higher levels of hope may play an important role in determining treatment retention. PMID- 21632189 TI - Isolation and characterization of emerging subgroup J avian leukosis virus associated with hemangioma in egg-type chickens. AB - Subgroup J avian leukosis virus (ALV-J), first isolated in 1989, predominantly causes myeloid leukosis (ML) in meat-type or egg-type chicken. Since 2006, the clinical cases of hemangioma rather than ML in commercial layer flocks associated with ALV-J have been reported, but it was still not clear whether the novel oncogenic ALV-J had emerged. We characterized SCAU-HN06 isolate of ALV-J from hemangioma in commercial Roman layers through animal experiment and full-length proviral genome sequence analysis. The SPF white leghorn egg-type chickens infected with SCAU-HN06 in ovo at day 11 of incubation showed an overall incidence of 56% hemangioma and 8% renal tumor throughout the 22-week trial, the mortality rate was 16%. Most genes of SCAU-HN06 isolate showed high nucleotide sequence identity to JS09GY6 which was isolated from Hy-Line Variety Brown layers suffering hemangioma. The 19-bp insertion in leader sequence and one key deletion in E element were the common features of SCAU-HN06 and JS09GY6. SCAU-HN06 and those ALV-Js associated with hemangioma, possibly recombinants of ALV-J and other avian retrovirus, may share the same ancestor. PMID- 21632190 TI - Effects of the removal of cytoplasm on the development of early cloned bovine embryos. AB - Oocyte cytoplasm plays a prominent role in cloned embryonic development. To investigate the influence of oocyte cytoplasmic amount on cloned embryo development, we generated bovine somatic cell nuclear transfer (SCNT) embryos containing high (30-40% of the cytoplasm was removed), medium (15-25% of the cytoplasm was removed) and low (<10% of the cytoplasm was removed) nucleocytoplasmic volume ratios (N/C) using enucleated metaphase II oocyte as recipient, and fibroblast as donor nucleus, and analyzed the expression levels of ND1, Cytb and ATPase6, as well as the embryonic quality. The results indicated: (1) the process of embryonic development was not influenced by <40% of cytoplasm removal; (2) the rate of blastocyst formation, the total number of blastomere and the ratio of ICM to TE were inversely proportional to the N/C; (3) SCNT embryos with reduced volume equal to 75-85% or >90% of an intact oocyte volume showed similar karyotype structure of the donor cells; (4) the number of mtDNA copy was larger in low N/C embryos than that in medium or high N/C embryos, and the expression levels of each gene hardly varied from the 2-cell to 8-cell stage, while the expression levels increased dramatically at the blastocyst stage; (5) from 16-cell to the blastocyst stage, the change of the expression level of each gene was not significant between low N/C embryos and IVF embryos, but it was more significant than those of high or medium N/C embryos. The results suggest that the decrease of mtDNA copy number and mitochondrial gene expression may be related to the impairment in early embryonic development, and removal of <10% adjacent cytoplasm volume may be optimal for bovine SCNT embryo development. PMID- 21632191 TI - Decreased resting energy expenditure in patients with Duchenne muscular dystrophy. AB - BACKGROUND: Skeletal muscle metabolism is a major determinant of resting energy expenditure (REE). Although the severe muscle loss that characterizes Duchenne muscular dystrophy (DMD) may alter REE, this has not been extensively investigated. METHODS: We studied REE in 77 patients with DMD ranging in age from 10 to 37 years using a portable indirect calorimeter, together with several clinical parameters (age, height, body weight (BW), body mass index (BMI), vital capacity (VC), creatine kinase, creatinine, albumin, cholinesterase, prealbumin), and assessed their influence on REE. In addition, in 12 patients maintaining a stable body weight, the ratio of energy intake to REE was calculated and defined as an alternative index for the physical activity level (aPAL). RESULTS: REE (kcal/day, mean+/-SD) in DMD patients was 1123 (10-11 years), 1186+/-188 (12-14 years), 1146+/-214 (15-17 years), 1006+/-136 (18-29 years) and 1023+/-97 (>=30 years), each of these values being significantly lower than the corresponding control (p<0.0001). VC (p<0.001) was the parameter most strongly associated with REE, followed by BMI (p<0.01) and BW (p<0.05). The calculated aPAL values were 1.61 (10-11 years), 1.19 (12-14 years), 1.16 (15-17 years), and 1.57 (18-29 years). CONCLUSION: The REE in DMD patients was significantly lower than the normal value in every age group, and strongly associated with VC. Both the low REE and PAL values during the early teens, resulting in a low energy requirement, might be related to the obesity that frequently occurs in this age group. In contrast, the high PAL value in the late stage of the disease, possibly due to the presence of respiratory failure, may lead to a high energy requirement, and thus become one of the risk factors for development of malnutrition. PMID- 21632192 TI - ProstAtlas: a digital morphologic atlas of the prostate. AB - Computer-aided medical interventions and medical robotics for prostate cancer have known an increasing interest and research activity. However before the routine deployment of these procedures in clinical practice becomes a reality, in vivo and in silico validations must be undertaken. In this study, we developed a digital morphologic atlas of the prostate. We were interested by the gland, the peripheral zone and the central gland. Starting from an image base collected from 30 selected patients, a mean shape and most important deformations for each structure were deduced using principal component analysis. The usefulness of this atlas was highlighted in two applications: image simulation and physical phantom design. PMID- 21632193 TI - Comparison of the accuracy of subtraction CT angiography performed on 320 detector row volume CT with conventional CT angiography for diagnosis of intracranial aneurysms. AB - OBJECTIVE: The accuracy of diagnosis of intracranial aneurysms by subtraction computed tomography angiography (CTA) was compared with conventional non subtracted CTA and with digital subtraction angiography (DSA). METHODS: 56 patients with spontaneous subarachnoid hemorrhage (SAH) and suspected intracranial aneurysms were evaluated from September 2009 to January 2010. All underwent 320-detector row volume CT-CTA examinations. Non-contrast CT of each patient's head with the same scan range was performed before the routine CTA scan as the mask image for subtraction. The subtraction CTA volume data was obtained by subtracting the mask image volume data from the conventional non-subtracted CTA volume data. Subtraction and conventional CTA volume data were transmitted to a VOXAR workstation and two physicians with experience in diagnostic imaging of the nervous system independently carried out image post-processing and judged the results. Neurosurgeons performed endovascular treatment or surgical clipping based on information available through the CTA alone. RESULTS: In 42 patients, 51 aneurysms were detected by DSA. On a per-aneurysm basis, the diagnostic sensitivity of subtraction CTA was 98.9% for physician 1 and 100% for physician 2. The sensitivity of conventional CTA was 93.7% for physician 1 and 92.6% for physician 2. There was excellent inter-observer agreement (kappa=0.84, 95% confidence interval 0.82-0.85). The overall sensitivity, specificity, positive predictive and negative predictive values of subtraction CTA were all 100%. The overall sensitivity, specificity, positive predictive and negative predictive values of non-subtracted CTA were 94%, 100%, 100% and 76%, respectively. Therapeutic decisions could be made for all 42 patients based on subtraction CTA images, whereas conventional non-subtracted CTA provided sufficient information to make therapeutic decisions for only 35 patients. CONCLUSION: Conventional CTA has lower sensitivity for the detection of very small aneurysms and aneurysms adjacent to the skull when compared to subtraction CTA. Subtraction CTA performed on a 320-detector row volume CT is an accurate diagnostic tool that provides data equivalent to that obtained with three-dimensional-DSA for the detection of intracranial aneurysms. PMID- 21632194 TI - Experimental designs and their recent advances in set-up, data interpretation, and analytical applications. AB - In this review, the set-up and data interpretation of experimental designs (screening, response surface, and mixture designs) are discussed. Advanced set ups considered are the application of D-optimal and supersaturated designs as screening designs. Advanced data interpretation approaches discussed are an adaptation of the algorithm of Dong and the estimation of factor effects from supersaturated design results. Finally, some analytical applications in separation science, on the one hand, and formulation-, product-, or process optimization, on the other, are discussed. PMID- 21632195 TI - Validation parameters cannot be obtained without using pure substance. PMID- 21632196 TI - Mixed methods evaluation: pharmacists' experiences and beliefs toward an interactive communication approach to patient interactions. AB - OBJECTIVE: To characterize pharmacists' experience and explore their beliefs toward an interactive communication technique, the three prime questions (3PQs),where pharmacists ask about patients' understanding of medication's purpose, directions, and monitoring. METHODS: Mixed method design. Pharmacists were briefly trained and then integrated the 3PQs into their practice for two weeks. Pharmacists recorded their perceptions of patient interactions, completed a survey addressing self-efficacy and role beliefs toward the 3PQs, and participated in a focus group. RESULTS: Eleven pharmacists participated and the 3PQs were used with 176 patients. Most interactions were under 5min. Pharmacists reported that questions about directions and monitoring were most effective in gathering new information with refills whereas medication purpose question was preferred for new fills. The majority of pharmacists were certain they could use the 3PQs and agreed it was their role. Five themes arose from the qualitative analysis: established communication routines, enhanced patient-pharmacist relationships, good medication history, tailoring of the 3PQs, and impact of pharmacy organization. CONCLUSION: The 3PQs enabled pharmacists to briefly assess patient medication experiences and tailor education while fostering patient centered relationships in pharmacy practice. PRACTICE IMPLICATIONS: While the 3PQs may enhance pharmacists' patient assessment; integration may challenge pharmacists' work routine. PMID- 21632197 TI - Who uses online interventions for problem drinkers? AB - The goal of this research was to understand why some people use online interventions for drinking problems, whereas others with comparable access to the interventions do not. As part of a randomized controlled trial, 92 participants in the experimental condition were provided access to a password-protected version of a Web-based personalized feedback intervention (the Check Your Drinking [CYD] screener, www.CheckYourDrinking.net). Information collected at baseline was compared between those who accessed the Web site and those who did not. Those who accessed the Web site tended to be more frequent users of the Internet, to drink less, and to perceive that others of the same age and gender drank less as compared with those who did not access the intervention. Some of these results are troubling as the preferred target of this type of intervention would be those who drink more and perceive that others are also heavy alcohol consumers. PMID- 21632198 TI - Frontline counselors in organizational contexts: a study of treatment practices in community settings. AB - This study addresses the challenge of implementing evidence-based treatment approaches in typical community settings. It identifies individual and organizational characteristics associated with two contrasting treatment approaches used by frontline practitioners. One treatment approach involves techniques supported by research; the other approach involves techniques primarily supported by experience and tradition. The study uses a nested probability sample of 45 organizations and 279 frontline practitioners. Multilevel (hierarchical linear modeling) regression models appropriately address the nested sample. The findings indicate that practitioner beliefs and components of organizational social contexts are associated with treatment approach. The use of an evidence-supported treatment approach is associated with opportunities to use training and with transformational leadership. A traditional treatment approach is more commonly used when practitioners have more positive perceptions of the organizational climate. The findings underscore the challenge of implementing evidence-based treatment techniques among counselors committed to traditional approaches. PMID- 21632199 TI - Determining the efficacy of auricular acupuncture for reducing anxiety in patients withdrawing from psychoactive drugs. AB - Auricular acupuncture (AA) is a widely accepted treatment option for substance abuse that is used in more than 700 treatment centers worldwide. Despite claims of perceived clinical benefits by patients and treatment staff, research efforts have failed to substantiate purported benefits, and the mechanism(s) by which AA serves in the treatment of addiction remain inconclusive. Numerous studies have shown AA to be an effective treatment for perioperative anxiety. In this study, we hypothesize that AA reduces the anxiety associated with withdrawal from psychoactive drugs. The study used a randomized, controlled design and included a sample of 101 patients recruited from an addiction treatment service. Subjects were assigned to one of three treatment groups (National Acupuncture Detoxification Association [NADA] AA, AA at sham points, or treatment setting control) and were instructed to attend treatment sessions for 3 days. The primary outcome measure state anxiety was assessed using a pretest-posttest treatment design. The study hypothesis was not confirmed. The NADA protocol was not more effective than sham or treatment setting control in reducing anxiety. The widespread acceptance of AA in the treatment of addiction remains controversial. PMID- 21632200 TI - MicroRNAs in colorectal cancer: function, dysregulation and potential as novel biomarkers. AB - BACKGROUND: MicroRNAs (miRNAs) are short non-coding segments of RNA which are involved in normal cellular development and proliferation. Recent studies have identified altered miRNA expression in both tumour tissues and circulation in the presence of colorectal cancer. These altered expression patterns may serve as novel biomarkers for colorectal cancer. This review explores recent developments in this rapidly evolving field. METHODS: A thorough literature search was performed to identify studies describing miRNA expression in colorectal cancer. Specific areas of interest included miRNA expression patterns in relation to development, diagnosis, progression and recurrence of disease, and potential future therapeutic applications. RESULTS: MiRNAs are associated with the development and progression of colorectal cancer. These may be either overexpressed or underexpressed (depending on the specific miRNA). Although there are fewer published studies regarding circulating miRNAs, these appear to be reflective of alterations in tissue expression and may have a potential role as minimally invasive biomarkers. CONCLUSION: MiRNAs have immense potential for refinement of the current processes for diagnosis, staging and prognostic prediction. They may also provide potential future therapeutic targets in the management of colorectal cancer. PMID- 21632201 TI - [Perianaesthetic concerns for the new robot-assisted transaxillary thyroid surgery: a report of seven first cases]. AB - The gasless transaxillary robot-assisted endoscopic thyroid surgery is recently proposed and developed in South Corea and USA. We reported the perianaesthestic concerns for the seven first patients scheduled to undergo this innovative surgical technique in France. The anaesthetic considerations focused on the length of surgery according to the learning curve, the risk of the arm posture and the postoperative painful evaluation and relief. PMID- 21632202 TI - [Good improvement of intubation conditions in a patient with acromegaly]. PMID- 21632203 TI - [Acute non-obstetrical diseases during pregnancy and role of the obstetrician]. AB - The management of a pregnant woman with an acute non-obstetrical disease must be made in narrow collaboration with an obstetrician. This one must be warned from the beginning of the care of the patient. In a pregnant woman, any acute medical, surgical or traumatic non-obstetrical disease can have obstetrical consequences. The diagnostic and therapeutic management of an acute non-obstetrical disease can have iatrogenic consequences during pregnancy. The most often described risks are early pregnancy loss, intra-uterine fetal death, placenta abruption, direct fetal hurts, preterm labor, prematurity and its complications. Obstetrical complications can induce maternal and neonatal life-threatening risks. Simple and easily accessible examinations in emergency allow detecting the obstetrical consequences of an acute non-obstetrical disease. During the management of an acute non-obstetrical disease in a pregnant woman, the induced obstetrical consequence of the disease can require emergency action of the obstetrician in conditions associated with maternal life-threatening risk. During the management of an acute non-obstetrical disease in a pregnant woman, once the mother condition was stabilized, the obstetrician had to estimate the fetal consequences and to adapt his or her therapeutic attitude. He or she sets up the fetal and placental surveillance adapted to the obstetrical risks and decides on the duration of this surveillance. PMID- 21632204 TI - Testing a cognitive model of generalized anxiety disorder in the eating disorders. AB - Generalized anxiety disorder (GAD) is one of the most common comorbid disorders found in individuals with eating disorders. Despite this, little is known of shared vulnerability factors between the two disorders. The aim of the present study was to examine the four main components of a cognitive model for GAD in the eating disorders. One hundred and sixty-two females took part. Three groups were formed comprising of 19 participants with an eating disorder and GAD, 70 with an eating disorder without GAD and 73 healthy controls. All completed self-report questionnaires that measured eating attitudes, levels of GAD, intolerance of uncertainty, positive beliefs about worry, negative problem orientation, and cognitive avoidance. Participants with an eating disorder and GAD scored the highest on all four components when compared to healthy individuals and on most components when compared to those with an eating disorder. Participants with an eating disorder without GAD scored higher on all components compared to healthy controls. Findings extend our understanding of shared vulnerability factors between the eating disorders and GAD. PMID- 21632205 TI - Blood, AIDS, and bureaucracy: the crisis and the tragedy. AB - The politics of health were never tested more than when AIDS surfaced at the beginning of the 1980s in the industrialized nations. In those countries, it became the most important medical crisis of the last half of the 20th century. Today, the significance of AIDS remains as not only an unrelenting disease but also as a disease that continues to affect social and political life throughout the entire world. The connection between blood transfusion and AIDS is now under control in the industrialized countries but only because of lessons that took too long to learn over the past 25 years. That process had different roots and effects depending on the various national blood programs and policies in different countries. That is illustrated by comparing events in France, Japan, Canada, and the United States that differed in donor and patient populations and on decisions made and secrets kept. Some of the problems persist to this day in parts of the world. Overall, the lessons learned from what happened with blood early in the AIDS epidemic apply to other aspects of human disease and could help in facing the new problems that are sure to appear in the future. PMID- 21632206 TI - Leukocyte transfusion and the development of the continuous-flow blood cell separator. AB - The treatment of anemia and thrombocytopenia with allogeneic cell transfusions is an effective and well-developed technology. However, leukocyte replacement transfusion has been frustrated by the physiology of the leukocytes. To achieve effective leukocyte replacement, the continuous-flow centrifugal blood cell separator was developed, and it soon proved to be an important instrument for separation, collection, and transfusion of all the components of the blood. Thus, the continuous-flow centrifugal blood cell separator has become an important instrument in the science of blood collection and transfusion. PMID- 21632207 TI - Motor coordination and health-related physical fitness of children with developmental coordination disorder: a three-year follow-up study. AB - Health-related physical fitness is an important risk factor of cardiovascular disease. While previous studies have identified children with developmental coordination disorder (DCD) to be less physically fit than typically developing (TD) peers, there is limited longitudinal research in this area. This study was undertaken to evaluate concomitant changes in motor coordination and health related physical fitness of Taiwanese children with and without DCD over a three year period. The Movement Assessment Battery for Children (Movement ABC) test was used to evaluate motor coordination, while health-related physical fitness included several core components: (1) body mass index (BMI), (2) sit and reach forward, (3) long jump, (4) sit-ups, and (5) 800-m run. Both the Movement ABC and fitness tests were implemented once each a year for three years. Twenty-five children with DCD and 25 TD children, matched by age and gender participated in this study. The TD group showed significant long-term changes in BMI and long jump while the DCD group showed significant increases in BMI values and decreases in flexibility, measured by the sit and reach task. In general, children with DCD performed worse on the items of flexibility, muscle strength and muscle endurance after the first year. Compared to age- and gender-matched norms, children with DCD not only were less physically fit, but showed a significant long-term decline in flexibility and abdominal or core strength (sit-ups). In years two and three, there was a significant negative correlation between poor fitness and motor coordination. Based on the results of this longitudinal study, greater attention should be paid to monitoring and improving physical fitness of children with DCD to prevent further health-related problems while intervention. PMID- 21632208 TI - Pathogenesis of systemic lupus erythematosus revisited 2011: end organ resistance to damage, autoantibody initiation and diversification, and HLA-DR. AB - Systemic lupus erythematosus (SLE) is a multi-system disorder resulting from interaction of susceptibility genes and environmental factors. SLE has protean clinical presentations at the initial diagnosis and relapses. SLE-related autoantibodies have unique patterns of diversification to linked proteins such as the snRNP particle and the diversification takes years before clinical diagnosis. There are both clinical and experimental evidence to indicate that separate genes contribute to autoimmunity and end organ damage and these genes are independent and interactive. Among the numerous susceptibility genes, HLA-D complex is dominant. Results from the authors' laboratories led us to postulate a unified hypothesis for SLE pathogenesis. This hypothesis states that SLE-autoantibodies are initiated by environmental T cell epitope mimics of the SLE-related autoantigens in hosts with susceptible HLA-D alleles. These autoantibodies diversify over a period of years due the accumulation of cross-reactive T cells. This process ultimately leads to the generation of organ specific autoantibodies and autoreactive effector T cells due to the polyreactive nature of T and B cell receptors from hosts with susceptibility genes to end organ damage, resulting in protean clinical presentations. This hypothesis accounts for most of the features unique to SLE and has clinical implications as to how patients should be treated. PMID- 21632209 TI - Cellular microRNAs (miRNAs) and Sjogren's syndrome: candidate regulators of autoimmune response and autoantigen expression. AB - MicroRNAs (miRNAs) are small non-coding RNA molecules that suppress gene expression at post-transcriptional level. miRNAs are considered as fine-tuning regulators of diverse biological processes, including the development and function of the immune system. Emerging data have implicated the deregulated expression of certain miRNAs or miRNA networks in the pathogenesis of autoimmune diseases. Sjogren's syndrome (SS) is a common chronic autoimmune disease, characterized by destruction and dysfunction of the exocrine glands (predominantly of the salivary and lachrymal glands). Humoral autoimmune responses observed in the disease, primarily target Ro/SSA and La/SSB ribonucleoproteins, whilst aberrantly increased expression of these autoantigens has been described in the salivary glands (SG) and the salivary gland epithelial cells (SGEC) of SS patients. Comparative array analysis of miRNA expression in the SGs of SS and control subjects had revealed distinctive miRNA signatures in SS patients, associated with glandular inflammation and dysfunction. Furthermore, the expression analysis of miRNAs that are predicted to target Ro/SSA and La/SSB autoantigens revealed differential expression of certain miRNAs in the SG tissues, SGECs and peripheral blood mononuclear cells (PBMC) of SS patients and controls. Although these association data implicate miRNAs in SS pathogenesis, thorough functional studies are needed to delineate their role in disease. PMID- 21632211 TI - Advancement genioplasty in Class I patients: predictability and stability of facial profile changes. AB - This retrospective study evaluated the skeletal and soft tissue facial profile changes as well as the predictability and the short-term stability of the soft tissue response to advancement genioplasty in Class I dental arch relationship patients. The study included 14 adult patients who presented a Class I dental arch but a Class II skeletal arch relationship and underwent advancement genioplasty exclusively. Lateral cephalograms taken immediately preoperatively (T1), immediately postoperatively (T2) and 1 year postoperatively (T3) were analysed. The hard tissue pogonion was sagittally advanced by an average of 7.9 mm (p<0.001) (T1-T2). The soft tissue chin followed the sagittal skeletal chin movement and exceeded chin advancement due to the initial soft tissue swelling. In the vertical dimension, the skeletal chin moved 3.0mm (p<0.01) upwards whilst the soft tissue chin moved only 2.1mm upwards (p<0.01). All profile convexity angles increased significantly (p<0.001), implying that the profile was straightened by the advancement of the chin. In the short term, advancement genioplasty was a predictable and stable procedure for chin correction. A ratio of 1:1 may be used to predict the sagittal soft tissue to bony movements for the period from before to 1 year after surgery. PMID- 21632212 TI - The association of cardiac asystole with partial seizures: does it result from ictal or interictal activity? AB - Bradycardia or asystole that occur during some seizures may be life threatening as a leading cause of SUDEP. A patient with right and left temporal lobe onset seizures and preceding bardycardia or asystole is presented. He had bilateral hippocampal atrophy on MRI. The unreliability of ictal bradycardia or asystole as a lateralizing sign in patients with partial epilepsy and the role of interictal autonomic activity in heart rate changes during seizures are discussed. PMID- 21632210 TI - Role of HLA class II genes in susceptibility and resistance to multiple sclerosis: studies using HLA transgenic mice. AB - Multiple sclerosis (MS), an inflammatory and demyelinating autoimmune disease of CNS has both, a genetic and an environmental predisposition. Among all the genetic factors associated with MS susceptibility, HLA class II haplotypes such as DR2/DQ6, DR3/DQ2, and DR4/DQ8 show the strongest association. Although a direct role of HLA-DR alleles in MS have been confirmed, it has been difficult to understand the contribution of HLA-DQ alleles in disease pathogenesis, due to strong linkage disequilibrium. Population studies have indicated that DQ alleles may play a modulatory role in the progression of MS. To better understand the mechanism by which HLA-DR and -DQ genes contribute to susceptibility and resistance to MS, we utilized single and double transgenic mice expressing HLA class II gene(s) lacking endogenous mouse class II genes. HLA class II transgenic mice have helped us in identifying immunodominant epitopes of PLP in context of various HLA-DR and -DQ molecules. We have shown that HLA-DR3 transgenic mice were susceptible to PLP(91-110) induced experimental autoimmune encephalomyelitis (EAE), while DQ6 (DQB1*0601) and DQ8 (DQB1*0302) transgenic mice were resistant. Surprisingly DQ6/DR3 double transgenic mice were resistant while DQ8/DR3 mice showed higher disease incidence and severity than DR3 mice. The protective effect of DQ6 in DQ6/DR3 mice was mediated by IFNgamma, while the disease exacerbating effect of DQ8 molecule was mediated by IL-17. Further, we have observed that myelin-specific antibodies play an important role in PLP(91-110) induced EAE in HLA-DR3DQ8 transgenic mice. Based on these observations, we hypothesize that epistatic interaction between HLA-DR and -DQ genes play an important role in predisposition to MS and our HLA transgenic mouse model provides a novel tool to study the effect of linkage disequilibrium in MS. PMID- 21632213 TI - Are post intracerebral hemorrhage seizures prevented by anti-epileptic treatment? AB - Prophylactic antiepileptic treatment in patients with non-traumatic, non aneurysmatic spontaneous intracerebral hemorrhage (SICH) is controversial. The purpose of our study was to assess the occurrence of seizures and neurologic outcome in SICH patients who were treated with valproic acid or a placebo for a period of one month and follow-up of one year in a hospital inpatient neurologic department and ambulatory clinic settings. The study is a prospective randomized, double-blind, placebo-controlled clinical trial. The patients were treated for one month with either valproic acid (VPA) or placebo immediately after a SICH and were followed-up for one year to evaluate seizure rate and neurologic function as measured by the National Institutes of Health Stroke Scale (NIHSS). Seventy-two patients participated in the study--36 were treated with VPA and 36 with placebo. During follow-up, 21% of the patients developed seizures. A by-treatment difference in incident seizures was not detected. However, a difference between reduction in early seizures and late one was observed in the VPA group. VPA treated patients exhibited improved neurological outcome as measured by NIHSS. Early prophylaxis with VPA in SICH patients did not prevent the occurrence of seizures post intracerebral hemorrhage, but was found to reduce early seizures. VPA-treated patients had improved NIHSS scores, suggesting that this treatment may confer some neuroprotective effect. Further studies with a larger number of patients and with other antiepileptic drugs are needed to properly clarify this finding. PMID- 21632214 TI - Trends in the use of antiepileptic drugs in Taiwan from 2003 to 2007: a population-based national health insurance study. AB - PURPOSE: The aim of this study was to evaluate the prevalence of prescription and use of antiepileptic drugs (AEDs) for the treatment of epilepsy and other indications in a nationwide population using a prescription database. MATERIALS AND METHODS: AED prescription data were collected from the National Health Insurance Research Database (NHIRD) in Taiwan for a 5-year period (2003-2007). Patients prescribed AEDs at least two times from 2003 to 2007 were selected for the study from a random sample that included approximately 600,000 people. RESULTS: The prevalence of AED use (per 1000 inhabitants) increased from 12.6 in 2003 to 13.8 in 2007. The prevalence of newer AED use increased progressively from 1.0 in 2003 to 3.8 in 2007, but the prevalence of older AED use decreased during this time (11.6-10). Carbamazepine and valproic acid were the most common AEDs used. Among the newer generation of AEDs, gabapentin was the most frequently used. Newer AEDs were used primarily to treat pain disorders. The primary class of drugs used to treat epileptic disorders was older AEDs. CONCLUSION: An increase in the use of AEDs was observed over a 5-year period in data collected from NHIRD. This might implicate the use of newer compounds at clinical practice not only increased in the treatment of epilepsy, but also in the conditions other than epilepsy especially pain disorders. PMID- 21632215 TI - Myoclonic astatic epilepsy and the use of the ketogenic diet. AB - Myoclonic astatic epilepsy (MAE) is a rare childhood generalized epilepsy syndrome of unknown incidence and etiology. Onset may be explosive with a myriad of different seizure types and children may become severely affected with an epileptic encephalopathy. This disorder may be particularly sensitive to the ketogenic diet (KD). This article will briefly review the background, diagnostic criteria's and our current information regarding the use of dietary therapies in MAE. PMID- 21632216 TI - Can insight be predicted in first-episode psychosis patients? A longitudinal and hierarchical analysis of predictors in a drug-naive sample. AB - Poor insight is a ubiquitous phenomenon in psychosis with great repercussions on clinical practise and the outcomes of patients. Poor insight comprises "state" and "trait" components. This paper targeted predictors of global insight and insight dimensions at baseline in the drug-naive status of first-episode psychosis patients and during a 6-month follow up after episode remission. Seventy-seven consecutive and previously unmedicated patients with first-episode schizophrenia-spectrum disorders (FESSD) completed baseline and 6-month insight, premorbid, symptomatological and neuropsychological assessments. Insight measures served as dependent variables for a set of hierarchical multiple regression models. Premorbid personality abnormalities and duration of untreated psychosis (DUP) significantly predicted 'state' and 'trait' insight global scores. Duration of untreated psychosis (DUP) significantly predicted 'state' insight, measured as refusal of treatment at baseline. Moreover, premorbid personality abnormalities and DUP with minor contributions of demographic variables, cognitive functioning and psychopathological dimensions predicted 'trait insight', defined as insight after remission of the psychosis episode 'Insight improver' FESSD patients showed better late adolescent premorbid adjustment, lower personality disturbances (sociopathic, schizoid and schizotypy dimensions), shorter DUP, and lower positive, negative and disorganisation symptoms and better cognitive performance on the Trail Making B test at the 6-month follow-up assessment. Premorbid personality abnormalities and DUP were predictors of 'state' and 'trait' insight, both at global scores and dimension levels. Moreover, insight improvement in patients with FESSD was related to premorbid abnormalities (in both adjustment and personality), shorter DUP, fewer positive and negative symptoms and better performance in cognitive tests at the 6-month follow up. PMID- 21632217 TI - Superior in vitro activity of ertapenem and piperacillin/tazobactam against recent clinical isolates of Proteus mirabilis from intra-abdominal infections (SMART 2009-2010). PMID- 21632218 TI - How are we assessing functioning in schizophrenia? A need for a consensus approach. AB - Efforts to improve everyday living skills are important in schizophrenia. What constitutes appropriate functioning is however difficult to evaluate. Several instruments were tested in different settings throughout Europe (EGOFORS initiative), including two new questionnaires (FROGS, PSRS), to provide directions on domains to be assessed and tools to be used. PMID- 21632219 TI - Effect of the structure of imidazolium cations in [BF4](-)-type ionic liquids on direct electrochemistry and electrocatalysis of horseradish peroxidase in Nafion films. AB - The direct electrochemistry and bioelectrocatalysis of horseradish peroxidase (HRP) in Nafion films at glassy carbon electrode (GCE) was investigated in three [BF(4)](-)-type room-temperature ionic liquids (ILs) to understand the structural effect of imidazolium cations. The three ILs are 1-ethyl-3-methylimidazolium tetrafluoroborate ([Emim][BF(4)]), 1-butyl-3-methylimidazolium tetrafluoroborate ([Bmim][BF(4)]) and 1-hexyl-3-methylimidazolium tetrafluoroborate ([Hmim][BF(4)]). A small amount of water in the three ILs is indispensable for maintaining the electrochemical activity of HRP in Nafion films, and the optimum water contents decrease with the increase of alkyl chain length on imidazole ring. Analysis shows that the optimum water contents are primarily determined by the hydrophilicity of ILs used. In contrast to aqueous medium, ILs media facilitate the direct electron transfer of HRP, and the electrochemical parameters obtained in different ILs are obviously related to the nature of ILs. The direct electron transfer between HRP and GCE is a surface-confined quasi reversible single electron transfer process. The apparent heterogeneous electron transfer rate constant decreases gradually with the increase of alkyl chain length on imidazole ring, but the changing extent is relatively small. The electrocatalytic reduction current of H(2)O(2) at the present electrode decreases obviously with the increase of alkyl chain length, and the mass transfer of H(2)O(2) via diffusion in ILs should be responsible for the change. In addition, the modified electrode has good stability and reproducibility; the ability to tolerate high levels of F(-) has been greatly enhanced due to the use of Nafion film. When an appropriate mediator is included in the sensing layer, a sensitive nonaqueous biosensor could be fabricated. PMID- 21632220 TI - Effects of lipid chain length on the surface properties of alkylaminomethyl rutin and of its mixture with model lecithin membrane. AB - Three model flavonoid-based bioactive molecules with different lipid chain lengths (RuCn: n=8, 12, 18) were newly synthesized. The surface properties [surface pressure (pi)-area (A), surface potential (DeltaV)-surface pressure (pi) and dipole moment (u(?))-surface pressure (pi)] of pure RuCn and the lecithin membrane compounds had been investigated by using the Langmuir monolayer technology. The results suggested that the distinctive monolayer behavior of RuCn is strongly dependent on the lipid chain length. The great differences in the monolayer properties brought by the lipid chain length could be attributed to two major factors: (i) the ionization degree of the bulky hydrophilic head group (including hydroxyl and NH groups) alters its local field solely via the surface potential; (ii) tring molecular (or dipole) packing density within monolayers. The excess Gibbs energy (DeltaG((ex))) calculated for the RuCn-lecithin mixed monolayers infers that higher stability of the mixed monolayer can be strengthened as the lipid chain length decreases. And the addition of RuCn into lecithin membrane may increase the total u(?) of the binary mixed monolayers, which could inhibit the hydration of the lecithin's hydrophilic head groups. The shorter the lipid chain length of RuCn (e.g., RuC8) is, the higher the surface activity can be. Our findings provide a molecular basis for the application of such class of biomolecules in the functional food, cosmetics and medicine. PMID- 21632221 TI - [Congenital toxoplasmosis following infection occurring late in pregnancy]. AB - Monthly serological screening of non immune pregnant women is recommended for prevention of congenital toxoplasmosis. However, this screening is often interrupted before delivery. We report a case of congenital toxoplasmosis following infection occurring late in pregnancy. This documented case highlights the need for a final routine serological test, 2-3 weeks post-partum for all seronegative pregnant women. In fact, the screening of congenital toxoplasmosis cases allows the early administration of specific treatment that avoids later severe complications such as chorioretinitis. PMID- 21632222 TI - Carnivorous planktonic Difflugia (Protista, Amoebina Testacea) and their predators. AB - Four planktonic species of Difflugia co-occurring in a south Chinese reservoir were found to be carnivorous, but the diet was widest in the largest species (D. tuberspinifera) and narrowest in the smallest (D. hydrostatica). It included rotifers, ciliates, dinoflagellates, floating eggs, and small particles associated with organic debris. Scavenging and cannibalism were also observed. Species with a collared test (D. biwae, D. mulanensis) showed a form of suction feeding, while species with teeth on the pseudostome used these, together with their pseudopods, as "inverted crown corks", providing leverage for opening the lorica of their (rotifer) prey. Predators of Difflugia included cyclopoid copepods. In addition, the rotifers Asplanchna priodonta, Ploesoma hudsoni and, occasionally, big ciliates (Stentor sp.) all ingested their prey as a whole. PMID- 21632223 TI - Ca(2+)-dependent in vivo protein phosphorylation and encystment induction in the ciliated protozoan Colpoda cucullus. AB - Encystment induction of Colpoda cucullus is promoted by an increase in external Ca(2+) and overpopulation of Colpoda vegetative cells. Using phos-tag detection assays, the present study revealed that the in vivo phosphorylation level in several proteins [33 kDa, 37 kDa, 37.5 kDa, 43 kDa, 47 kDa, 49 kDa, etc.] was raised when the vegetative cells were stimulated by overpopulation to encyst in a medium containing 0.1 mM Ca(2+) or without the addition of Ca(2+). Both overpopulation-mediated encystment induction and protein phosphorylation were suppressed by the addition of EGTA. Ca(2+)/overpopulation-stimulated encystment induction and protein phosphorylation were also suppressed by the addition of BAPTA-AM. These results suggest that the Ca(2+) inflow promoted by cell-to-cell stimulation due to overpopulation may activate signaling pathways involving protein phosphorylation and encystment induction. In the presence of cAMP-AM, the phosphorylation levels of 33 kDa, 37 kDa, 37.5 kDa, 43 kDa, 47 kDa and 49 kDa proteins were enhanced, and encystment induction was promoted. Enzyme immunoassays (EIAs) showed that intracellular cAMP concentration was raised prior to encystment when the cells were stimulated by overpopulation. These results suggest that cAMP/PKA-dependent protein phosphorylation, which is an event on Ca(2+)-triggered signaling pathways, may be involved in encystment induction. PMID- 21632224 TI - Characterization of a newly discovered Beauveria bassiana isolate to Franklimiella occidentalis Perganda, a non-native invasive species in China. AB - In this study, a new virulent Beauveria bassiana isolate (B. bassiana-CYT5) had been identified as a new member of the species B. bassiana. The B. bassiana-CYT5 isolate was compared with four other virulent B. bassiana isolates and found to be highly infectious and virulent against the Franklimiella occidentalis Perganda. The B. bassiana-CYT5 could approximately 93.08% mortality of F. occidentalis 6 days post inoculation in the concentration of 1*10(8) conidia/mL. The phylogenetic tree based on ITS and partial sequence of elongation factor 1 alpha (EF1-alpha) indicated that B. bassiana-CYT5 isolate was in a cluster of B. bassiana. Furthermore, B. bassiana-CYT5 isolate demonstrated high heat tolerance (60-100% relative germination) between 1-h and 2-h exposure at 37 degrees C, 38 degrees C, 39 degrees C and 40 degrees C, respectively. So our results suggested that B. bassiana-CYT5 isolate could be a new efficient biocontrol agent against F. occidentalis. PMID- 21632225 TI - The autotransporter protein from Bordetella avium, Baa1, is involved in host cell attachment. AB - Bordetella avium is a Gram negative upper respiratory tract pathogen of birds. B. avium infection of commercially raised turkeys is an agriculturally significant problem. Here we describe the functional analysis of the first characterized B. avium autotransporter protein, Baa1. Autotransporters comprise a large family of proteins found in all groups of Gram negative bacteria. Although not unique to pathogenic bacteria, autotransporters have been shown to perform a variety of functions implicated in virulence. To test the hypothesis that Baa1 is a B. avium virulence factor, unmarked baa1 deletion mutants (Deltabaa1) were created and tested phenotypically. It was found that baa1 mutants have wild-type levels of serum sensitivity and infectivity, yet significantly lower levels of turkey tracheal cell attachment in vitro. Likewise, semi-purified recombinant His-tagged Baa1, expressed in Escherichia coli, was shown to bind specifically to turkey tracheal cells via western blot analysis. Taken together, we conclude that Baa1 acts as a host cell attachment factor and thus plays a role B. avium virulence. PMID- 21632226 TI - Rapid and dissimilar response of ammonia oxidizing archaea and bacteria to nitrogen and water amendment in two temperate forest soils. AB - Biochemical processes relevant to soil nitrogen (N) cycling are performed by soil microorganisms affiliated with diverse phylogenetic groups. For example, the oxidation of ammonia, representing the first step of nitrification, can be performed by ammonia oxidizing bacteria (AOB) and, as recently reported, also by ammonia oxidizing archaea (AOA). However, the contribution to ammonia oxidation of the phylogenetically separated AOA versus AOB and their respective responsiveness to environmental factors are still poorly understood. The present study aims at comparing the capacity of AOA and AOB to momentarily respond to N input and increased soil moisture in two contrasting forest soils. Soils from the pristine Rothwald forest and the managed Schottenwald forest were amended with either NH(4)(+)-N or NO(3)(-)-N and were incubated at 40% and 70% water-filled pore space (WFPS) for four days. Nitrification rates were measured and AOA and AOB abundance and community composition were determined via quantitative PCR (qPCR) and terminal restriction length fragment polymorphism (T-RFLP) analysis of bacterial and archaeal amoA genes. Our study reports rapid and distinct changes in AOA and AOB abundances in the two forest soils in response to N input and increased soil moisture but no significant effects on net nitrification rates. Functional microbial communities differed significantly in the two soils and responded specifically to the treatments during the short-term incubation. In the Rothwald soil the abundance and community composition of AOA were affected by the water content, whereas AOB communities responded to N amendment. In the Schottenwald soil, by contrast, AOA responded to N addition. These results suggest that AOA and AOB may be selectively influenced by soil and management factors. PMID- 21632227 TI - The GlideScope in current clinical practice. PMID- 21632228 TI - Prophylactic glycopyrrolate prevents bradycardia after spinal anesthesia for Cesarean section: a randomized, double-blinded, placebo-controlled prospective trial with heart rate variability correlation. AB - STUDY OBJECTIVE: To determine if prophylactic glycopyrrolate prevents bradycardia after spinal anesthesia. DESIGN: Prospective, randomized, double-blinded, placebo controlled study. SETTING: Large university-affiliated community hospital. PATIENTS: 81 consecutive term parturients (not in active labor) who were scheduled for elective Cesarean section. INTERVENTIONS: Parturients received 1.0 to 1.5 L of intravenous Ringer's lactate and either glycopyrrolate 0.4 mg or an equal volume of saline, with caregivers blinded to the immediate sequelae of study drug. Each patient received intrathecal bupivacaine (12 to 14 mg) with morphine sulfate (0.1 to 0.2 mg). MEASUREMENTS: Continuous heart rate (HR) and blood pressure monitoring occurred for 20 minutes, with the minimum HR recorded for each 5-minute epoch. Heart rates < 60 beats per minute defined bradycardia. Heart rate variability (HRV) analysis occurred offline. MAIN RESULTS: None of 34 patients administered glycopyrrolate and 6 of 35 (17%) patients receiving saline experienced bradycardia (P = 0.02476). Time domain, frequency domain, and nonlinear and embedded spectrum entropy analyses all reflected the decrease in HRV accompanying administration of glycopyrrolate. CONCLUSION: Bradycardia after spinal anesthesia occurs commonly. Prophylactic glycopyrrolate may prevent the bradycardia, but not necessarily the hypotension. PMID- 21632229 TI - Is this complex regional pain syndrome or reflex sympathetic dystrophy? PMID- 21632230 TI - HRCT features of interstitial lung disease in dermatomyositis with anti-CADM-140 antibody. AB - BACKGROUND: Anti-CADM-140 antibody (anti-CADM-140), also referred to as anti melanoma differentiation-associated gene 5 (MDA5) antibody, is a myositis specific antibody identified in the sera of patients with clinically amyopathic dermatomyositis (C-ADM) and is associated with a worse prognosis in dermatomyositis-associated interstitial lung disease (DM-ILD). We sought to determine high-resolution computed tomography (HRCT) features of DM-ILD with anti CADM-140. METHODS: Twenty-five patients newly diagnosed with DM-ILD at Kyoto University Hospital between 2005 and 2009 were retrospectively reviewed. Serum anti-CADM-140 was measured in all patients at their first visit. Chest HRCT images taken prior to treatment were classified based on the dominant findings and their distribution, and compared between patients with and without the antibody. RESULTS: Of 25 DM-ILD patients, 12 were positive and 13 were negative for anti-CADM-140. HRCT patterns differed significantly between anti-CADM-140 positive and negative patients (P = 0.002). Lower consolidation or ground-glass attenuation (GGA) pattern (50.0%) and random GGA pattern (33.3%) were the predominant patterns in anti-CADM-140-positive cases, while lower reticulation pattern (69.2%) was frequently seen in anti-CADM-140-negative cases. Anti-CADM 140-positive cases were also significantly characterized by the absence of intralobular reticular opacities (0% in anti-CADM-140 (+) vs. 84.6% in anti-CADM 140 (-), P < 0.0001). CONCLUSIONS: Anti-CADM-140-positive DM-ILD was characterized by lower consolidation or GGA pattern, random GGA pattern, and the absence of intralobular reticular opacities. PMID- 21632231 TI - Cell invasion through basement membrane: the anchor cell breaches the barrier. AB - Cell invasion through basement membrane (BM) is a specialized cellular behavior critical to many normal developmental events, immune surveillance, and cancer metastasis. A highly dynamic process, cell invasion involves a complex interplay between cell-intrinsic elements that promote the invasive phenotype, and cell cell and cell-BM interactions that regulate the timing and targeting of BM transmigration. The intricate nature of these interactions has made it challenging to study cell invasion in vivo and model in vitro. Anchor cell invasion in Caenorhabditis elegans is emerging as an important experimental paradigm for comprehensive analysis of BM invasion, revealing the gene networks that specify invasive behavior and the interactions that occur at the cell-BM interface. PMID- 21632232 TI - G-quadruplex DNAzyme-based microcystin-LR (toxin) determination by a novel immunosensor. AB - In this paper, we demonstrate the application of versatile G-quadruplex-hemin DNAzymes in an immunoassay for detecting Microcystin-LR (MC-LR). Taking advantage of the high peroxidase activity of G-quadruplex-hemin complexes and the enhancement effect of gold nanoparticles (AuNPs), the method showed simple, high sensitive and selectivity detection of target toxin residues in water samples. The coated antigen, MC-LR-ovalbumin (OVA) coated on a plate, competed for MC-LR antibody with added target analyte to form antibody-antigen immune complexes. Subsequently, the immune complex reacted with G-quadruplex-labeled secondary antibodies for colorimetric detection of MC-LR. This assay specifically determined MC-LR in the linear range of 0.1-10 ng/ml, with a limit of detection (LOD) of 0.05 ng/mL for MC-LR. The results indicated that the novel immunoassay was an alternative to traditional plate-based immunoassay for MC-LR residue screening due to this method met the standard of World Health Organization (WHO) requirements for MC-LR content in drinking water (1 ng/mL). PMID- 21632233 TI - Bifunctional electro-optical nanoprobe to real-time detect local biochemical processes in single cells. AB - A bifunctional electro-optical nanoprobe with integrated nanoring electrode and optical nanotip was fabricated and investigated to simultaneously detect both electrical and optical signals in real-time with high spatial resolution. Concurrent measurements of the oxidant generation and the intracellular antioxidant levels in single cells correlate the stronger oxidant generation with an altered initial antioxidant response in the breast cancer cells in comparison to the normal ones suggesting that the cell malignancy is associated with the strength of oxidative stress, and the higher antioxidant level may be the cause of the drug resistance. While the optical detection indicates the fluctuation of the intracellular redox homeostasis, the chronoamperometric signals allow quantitative real-time detection of the H2O2 release and decay. Furthermore, the nanoscale probe enables localized simultaneous detections thus discovering that activated enzymes responsible for the oxidative stress target at specific membrane regions. This method promises applications in study of the dynamics of important physiological processes, and provides the opportunity to unravel the interplay of various signaling pathways. PMID- 21632234 TI - Microfluidic electrochemical immunoarray for ultrasensitive detection of two cancer biomarker proteins in serum. AB - A microfluidic electrochemical immunoassay system for multiplexed detection of protein cancer biomarkers was fabricated using a molded polydimethylsiloxane channel and routine machined parts interfaced with a pump and sample injector. Using off-line capture of analytes by heavily-enzyme-labeled 1 MUm superparamagnetic particle (MP)-antibody bioconjugates and capture antibodies attached to an 8-electrode measuring chip, simultaneous detection of cancer biomarker proteins prostate specific antigen (PSA) and interleukin-6 (IL-6) in serum was achieved at sub-pg mL-1 levels. MPs were conjugated with ~90,000 antibodies and ~200,000 horseradish peroxidase (HRP) labels to provide efficient off-line capture and high sensitivity. Measuring electrodes feature a layer of 5 nm glutathione-decorated gold nanoparticles to attach antibodies that capture MP analyte bioconjugates. Detection limits of 0.23 pg mL-1 for PSA and 0.30 pg mL-1 for IL-6 were obtained in diluted serum mixtures. PSA and IL-6 biomarkers were measured in serum of prostate cancer patients in total assay time 1.15 h and sensor array results gave excellent correlation with standard enzyme-linked immunosorbent assays (ELISA). These microfluidic immunosensors employing nanostructured surfaces and off-line analyte capture with heavily labeled paramagnetic particles hold great promise for accurate, sensitive multiplexed detection of diagnostic cancer biomarkers. PMID- 21632235 TI - Insect olfactory coding and memory at multiple timescales. AB - Insects can learn, allowing them great flexibility for locating seasonal food sources and avoiding wily predators. Because insects are relatively simple and accessible to manipulation, they provide good experimental preparations for exploring mechanisms underlying sensory coding and memory. Here we review how the intertwining of memory with computation enables the coding, decoding, and storage of sensory experience at various stages of the insect olfactory system. Individual parts of this system are capable of multiplexing memories at different timescales, and conversely, memory on a given timescale can be distributed across different parts of the circuit. Our sampling of the olfactory system emphasizes the diversity of memories, and the importance of understanding these memories in the context of computations performed by different parts of a sensory system. PMID- 21632237 TI - A systematic review of cancer related patient outcomes after anterior resection and abdominoperineal excision for rectal cancer in the total mesorectal excision era. AB - PURPOSE: It is a widely held view that anterior resection (AR) for rectal cancer is an oncologically superior operation to abdominoperineal excision (APE). However, some centres have demonstrated better outcomes with APE. We conducted a systematic review of high-quality studies within the total mesorectal excision (TME) era comparing outcomes of AR and APE. METHODS: A literature search was performed to identify studies within the TME era comparing AR and APE with regard to the following: circumferential resection margin (CRM) status, tumour perforation rates, specimen quality, local recurrence, overall survival (OS; 3 or 5 year), cancer-specific survival (CSS) and disease-free survival (DFS). Additional data regarding patient demographics and tumour characteristics was collected. RESULTS: Twenty four studies fulfilled the eligibility criteria with Newcastle-Ottawa scores of six or greater. Where a significant difference was found, all studies reported lower and more advanced tumours for APE and 4/5 studies observed more frequent use of neoadjuvant and adjuvant therapies in APE patients. Tumour perforation rates and CRM involvement where reported, were significantly greater for APE. 8 out of 10 studies showing significant differences in local recurrence reported higher rates for APE but no differences were observed with distant recurrence. Where differences were noted, AR was reported to have increased DFS, CSS and OS compared to APE. CONCLUSIONS: Patients treated with AR have lower rates of tumour perforation and CRM involvement and tend to have better outcomes with regard to disease recurrence and survival. However, tumours treated by APE are lower and more locally advanced. PMID- 21632236 TI - Glutamatergic signaling by midbrain dopaminergic neurons: recent insights from optogenetic, molecular and behavioral studies. AB - Although the notion that dopaminergic neurons utilize glutamate as a co transmitter has long been supported by tantalizing molecular, immunocytochemical and electrophysiological evidence it has only been with the recent addition of optogenetic and other approaches that the existence and functional relevance of this mechanism could be unambiguously demonstrated. Here we discuss the possible mechanisms of action of glutamate released from mesoaccumbens dopaminergic neurons based on recently accumulated evidence. Surprisingly, rather then to confirm a role in conventional fast excitatory transmission, the latest evidence suggests that glutamate released from dopaminergic neurons may primarily act through different unconventional presynaptic and postsynaptic mechanisms. PMID- 21632238 TI - Manganese biomining: A review. AB - Biomining comprises of processing and extraction of metal from their ores and concentrates using microbial techniques. Currently this is used by the mining industry to extract copper, uranium and gold from low grade ores but not for low grade manganese ore in industrial scale. The study of microbial genomes, metabolites and regulatory pathways provide novel insights to the metabolism of bioleaching microorganisms and their synergistic action during bioleaching operations. This will promote understanding of the universal regulatory responses that the biomining microbial community uses to adapt to their changing environment leading to high metal recovery. Possibility exists of findings ways to imitate the entire process during industrial manganese biomining endeavor. This paper reviews the current status of manganese biomining research operations around the world, identifies factors that drive the selection of biomining as a processing technology, describes challenges in exploiting these innovations, and concludes with a discussion of Mn biomining's future. PMID- 21632239 TI - A biochemically structured model for ethanol fermentation by Kluyveromyces marxianus: A batch fermentation and kinetic study. AB - Anaerobic batch fermentations of ricotta cheese whey (i.e. containing lactose) were performed under different operating conditions. Ethanol concentrations of ca. 22g L(-1) were found from whey containing ca. 44g L(-1) lactose, which corresponded to up to 95% of the theoretical ethanol yield within 15h. The experimental data could be explained by means of a simple knowledge-driven biochemically structured model that was built on bioenergetics principles applied to the metabolic pathways through which lactose is converted into major products. Use of the model showed that the observed concentrations of ethanol, lactose, biomass and glycerol during batch fermentation could be described within a ca. 6% deviation, as could the yield coefficients for biomass and ethanol produced on lactose. The model structure confirmed that the thermodynamics considerations on the stoichiometry of the system constrain the metabolic coefficients within a physically meaningful range thereby providing valuable and reliable insight into fermentation processes. PMID- 21632240 TI - A thermotolerant and cold-active mannan endo-1,4-beta-mannosidase from Aspergillus niger CBS 513.88: Constitutive overexpression and high-density fermentation in Pichia pastoris. AB - The mannan endo-1,4-beta-mannosidase gene man26A from Aspergillus niger CBS 513.88 was optimized according to the codon usage bias in Pichia pastoris and synthesized by splicing overlap extension PCR. It was successfully expressed in P. pastoris using constitutive expression vector pGAPzalphaA. The recombinant endo-beta-1,4-mannanase could work in an extremely board temperature range and over 30% relative activity were retained in the temperature range of 5-60 degrees C. The optimal pH value and temperature for activity were 5.0 and 45 degrees C, respectively. It was highly thermotolerant with a half-life time of 15min at 90 degrees C. A novel fed-batch strategy was developed successfully for high cell density fermentation and mannanase activity reached 5069U/mL after cultivation for 56h in 50L fermenter. The broad working temperature range, high thermotolerance and efficient expression made this enzyme possible to be applied in food, animal feed and the production of biofuels. PMID- 21632241 TI - Scale-up study of oxalic acid pretreatment of agricultural lignocellulosic biomass for the production of bioethanol. AB - Building on our laboratory-scale optimization, oxalic acid was used to pretreat corncobs on the pilot-scale. The hydrolysate obtained after washing the pretreated biomass contained 32.55g/l of xylose, 2.74g/l of glucose and low concentrations of inhibitors. Ethanol production, using Scheffersomyces stipitis, from this hydrolysate was 10.3g/l, which approached the predicted value of 11.9g/l. Diafiltration using a membrane system effectively reduced acetic acid in the hydrolysate, which increased the fermentation rate. The hemicellulose content of the recovered solids decreased from 27.86% before pretreatment to only 6.76% after pretreatment. Most of the cellulose remained in the pretreated biomass. The highest ethanol production after simultaneous saccharification and fermentation (SSF) of washed biomass with S. stipitis was 21.1g/l. PMID- 21632242 TI - Techno-economic implications of improved high gravity corn mash fermentation. AB - The performance of Saccharomyces cerevisiae MBG3964, a strain able to tolerate >18% v/v ethanol, was compared to leading industrial ethanol strain, Fermentis Ethanol Red, under high gravity corn mash fermentation conditions. Compared to the industrial ethanol strain, MBG3964 gave increased alcohol yield (140g L(-1) vs. 126g L(-1)), lower residual sugar (4g L(-1) vs. 32g L(-1)), and lower glycerol (11g L(-1) vs. 12g L(-1)). After 72h fermentation, MBG3964 showed about 40% viability, whereas the control yeast was only about 3% viable. Based on modelling, the higher ethanol tolerant yeast could increase the profitability of a corn-ethanol plant and help it remain viable through higher production, lower unit heating requirements and extra throughput. A typical 50M gal y(-1) dry mill ethanol plant that sells dried distiller's grain could potentially increase its profit by nearly $US3.4M y(-1) due solely to the extra yield, and potentially another $US4.1M y(-1) if extra throughput is possible. PMID- 21632243 TI - Methanesulfonamido-cyclohexylamine derivatives of 2,4-diaminopyrimidine as potent ALK inhibitors. AB - The incorporation of R,R-1,2-diaminocyclohexane at C4 in a series of 2,4 diaminopyrimidines led to a number of ALK inhibitors in which optimized activity was achieved by conversion of the 2-amino group into a methanesulfonamide. Tumor growth inhibition was observed when an orally bioavailable analog was evaluated in a Karpas-299 tumor xenograft mouse model. PMID- 21632244 TI - Synthesis and characterization of fluorinated homocysteine derivatives as potential molecular probes for 19F magnetic resonance spectroscopy and imaging. AB - A N-trifluoroacetyl-protected amino acid containing a thioester function, 2,2,2 trifluoro-N-(2-oxo-tetrahydrothiophen-3-yl)acetamide (TFA-tHcy), has been synthesized and characterized. It was then used to prepare a fluorine-labeled N homocysteinylated protein, (19)F-Hcy-epsilonN-Lys-albumin, that was characterized by SDS-PAGE, MALDI-TOF-MS, UV-vis and (19)F NMR spectroscopy. On average, four N trifluoroacetylhomocysteine residues were covalently conjugated to human serum albumin through the N-substituted homocysteine thiolactone. The in situ homocysteinylation of human plasma proteins with TFA-tHcy has also been performed and has led to the formation of N-homocysteinylated proteins, with albumin modification accounting for ca. 75% of all fluorine-labeled human plasma proteins. The synthesized fluorinated molecular probes can be potentially used as informative molecular probes for in vivo (19)F magnetic resonance spectroscopy and imaging. PMID- 21632245 TI - Selective accumulation of hybrid liposomes into adult T-cell leukemia cells along with induction of apoptosis. AB - Markedly inhibitory effects of hybrid liposomes (HL-n) composed of 90 mol% L alpha-dimyristoylphosphatidylcholine (DMPC) and 10 mol% polyoxyethylene(n) dodecyl ethers on the growth of adult T-cell leukemia cells were obtained for the first time. It is noteworthy that HL-n could selectively accumulate into the adult T-cell leukemia cells and induce apoptosis via caspase-3 activation. PMID- 21632246 TI - Unexpectedly convenient and stereoselective synthesis of 4alpha-acyloxy-2 chloropodophyllotoxins in the presence of BF3 .Et2O. AB - Twenty-one 4alpha-acyloxy-2-chloropodophyllotoxin derivatives (5a-u), whose C-4 spatial configuration was mainly stereocontrolled by the configuration of C-2 chlorine atom, were unexpectedly prepared by the reaction of 2 chloropodophyllotoxin with carboxylic acids in the presence of BF(3).Et(2)O. Compared with ordinary esterifications of carboxylic acids mediated by the condensation agent, for example, N,N'-diisopropylcarbodiimide (DIC), the present method made the procedure for the preparation of 4alpha-acyloxy-2 chloropodophyllotoxins more convenient, practical and easy. Meanwhile, the insecticidal activity of compounds 5a-u was preliminarily evaluated against the pre-third-instar larvae of Mythimna separata Walker in vivo at the concentration of 1mg/mL. PMID- 21632247 TI - Identification of new inhibitors of protein kinase R guided by statistical modeling. AB - We report the identification of new, structurally diverse inhibitors of interferon-induced, double-stranded RNA-activated protein kinase (PKR) using a combined experimental and computational approach. A training set with which to build a predictive statistical model was generated by screening a set of 80 known Ser/Thr kinase inhibitors against recombinant human PKR, resulting in the identification of 28 compounds from 18 chemical classes with <0.1 MUM <= IC(50) <= 20 MUM. The model built with this data was used to screen a database of 5 million commercially available compounds in silico to identify candidate inhibitors. Testing of 128 structurally diverse candidates resulted in the confirmation of 20 new inhibitors from 11 chemical classes with 2 MUM <= IC(50) <= 20 MUM. Testing of 34 analogs in the newly identified pyrimidin-2-amine active series provided initial SAR. One newly identified inhibitor, N-[2-(1H-indol-3 yl)ethyl]-4-(2-methyl-1H-indol-3-yl)pyrimidin-2-amine (compound 51), inhibited intracellular PKR activation in a dose-dependent manner in primary mouse macrophages without evident toxicity at effective concentrations. PMID- 21632248 TI - Evaluation of a cell penetrating prenylated peptide lacking an intrinsic fluorophore via in situ click reaction. AB - Protein prenylation involves the addition of either a farnesyl (C(15)) or geranylgeranyl (C(20)) isoprenoid moiety onto the C-terminus of many proteins. This natural modification serves to direct a protein to the plasma membrane of the cell. A recently discovered application of prenylated peptides is that they have inherent cell-penetrating ability, and are hence termed cell penetrating prenylated peptides. These peptides are able to efficiently cross the cell membrane in an ATP independent, non-endocytotic manner and it was found that the sequence of the peptide does not affect uptake, so long as the geranylgeranyl group is still present [Wollack, J. W.; Zeliadt, N. A.; Mullen, D. G.; Amundson, G.; Geier, S.; Falkum, S.; Wattenberg, E. V.; Barany, G.; Distefano, M. D. Multifunctional Prenylated Peptides for Live Cell Analysis. J. Am. Chem. Soc.2009, 131, 7293-7303]. The present study investigates the effect of removing the fluorophore from the peptides and investigating the uptake by confocal microscopy and flow cytometry. Our results show that the fluorophore is not necessary for uptake of these peptides. This information is significant because it indicates that the prenyl group is the major determinant in allowing these peptides to enter cells; the hydrophobic fluorophore has little effect. Moreover, these studies demonstrate the utility of the Cu-catalyzed click reaction for monitoring the entry of nonfluorescent peptides into cells. PMID- 21632249 TI - Inflammatory changes in infantile-onset LMNA-associated myopathy. AB - Mutations in LMNA cause wide variety of disorders including Emery-Dreifuss muscular dystrophy, limb girdle muscular dystrophy, and congenital muscular dystrophy. We recently found a LMNA mutation in a patient who was previously diagnosed as infantile onset inflammatory myopathy. In this study, we screened for LMNA mutations in 20 patients suspected to have inflammatory myopathy with onset at 2years or younger. The diagnosis of inflammatory myopathy was based on muscle pathology with presence of perivascular cuffing and/or endomysial/perimysial lymphocyte infiltration. We identified heterozygous LMNA mutations in 11 patients (55%), who eventually developed joint contractures and/or cardiac involvement after the infantile period. Our findings suggest that LMNA mutation should be considered in myopathy patients with inflammatory changes during infancy, and that this may help avoid life-threatening events associated with laminopathy. PMID- 21632250 TI - The SecY complex: conducting the orchestra of protein translocation. AB - Like the conductor of an orchestra, the Sec protein translocation channel is the platform needed to bring together the many different players required for the constitutive and obligatory process of protein transport. This conserved membrane channel, termed SecY in bacteria and Sec61 in eukaryotes, creates a ubiquitous protein-conducting pathway by which thousands of newly synthesized polypeptides make their way through the lipid bilayer. The channel is not a simple passive pore, however; it displays remarkable complexity by interacting with numerous soluble partners, including SecA, Syd, FtsY and the ribosome in bacteria. For several decades, scientists have been fascinated by the sophistication and versatility of this transport channel. In this review, we cover the current state of the field including some of the newest and most exciting findings on channel structure and mechanism of action. PMID- 21632251 TI - Reconstructing regulatory network transitions. AB - Cellular responses often involve a transition of cells from one state to another. A transition from a stem cell to a differentiated cell state, for example, might occur in response to gene expression changes induced by a transcription factor, or to signaling cascades triggered by a hormone or pathogen. Regulatory networks are thought to control such cellular transitions. Thus, many researchers are interested in reconstructing regulatory networks, not only with the aim of gaining a deeper understanding of cellular transitions, but also of using networks to predict and potentially manipulate cellular transitions and outcomes. In this review, we highlight approaches to the reconstruction of regulatory networks underlying cellular transitions, with special attention to transcriptional regulatory networks. We describe recent regulatory network reconstructions in a variety of organisms, and discuss the success they share in identifying new regulatory components, shared relationships and phenotypic outcomes. PMID- 21632252 TI - Coupling mitosis to DNA replication: the emerging role of the histone H4-lysine 20 methyltransferase PR-Set7. AB - To ensure accurate inheritance of genetic information through cell proliferation, chromosomes must be precisely copied only during S phase, and then correctly condensed and segregated during mitosis. Several new findings suggest that this tight coupling between DNA replication and mitosis is in part controlled by cell cycle regulated chromatin modifications, in particular due to the changing activity of lysine methyltransferase PR-Set7/SET8 that is responsible for the monomethylation of histone H4 at lysine 20. Cell cycle oscillation of PR-Set7 is orchestrated by ubiquitin-mediated proteolysis, and interference with this regulatory process leads to unscheduled licensing of replication origins and altered timing of mitotic chromosome compaction. This review provides an overview of how PR-Set7 regulates these two cell cycle events and highlights questions that remain to be addressed. PMID- 21632253 TI - Cdk5rap2 exposes the centrosomal root of microcephaly syndromes. AB - Autosomal recessive primary microcephaly (MCPH) is characterized by small brain size as a result of deficient neuron production in the developing cerebral cortex. Although MCPH is a rare disease, the questions surrounding its etiology strike at the core of stem cell biology. The seven genes implicated in MCPH all encode centrosomal proteins and disruption of the MCPH gene Cdk5rap2 in mice revealed its role in neural progenitor proliferation and in maintaining normal centriole replication control. We discuss here the impact that centrosome regulation has upon neural progenitors in the developing brain. We integrate the impact of centriole replication defects with the functions of Cdk5rap2 and other MCPH proteins, propose mechanisms for progenitor loss in MCPH, and discuss links to two other microcephaly syndromes. PMID- 21632255 TI - The biomechanical functional demand placed on knee and hip muscles of older adults during stair ascent and descent. AB - Age-related decline in physical capabilities may lead to older adults experiencing difficulty in performing everyday activities due to high demands placed on the muscles of their lower extremity. This study aimed to determine the biomechanical functional demand in terms of joint moments and maximal muscle capabilities at the knee and hip joints while older adults performed stair ascent (SA) and stair descent (SD). Eighty-four healthy older adults aged 60-88 years were tested. A torque dynamometer attached to a purpose-built plinth was utilized to measure muscle moments at the knee and hip joints. Participants also underwent full body 3-D biomechanical assessment of stair ascent and descent using an 8 camera VICON system (120Hz) with 3 Kistler force plates. Stair negotiation required knee extensor moments in excess of the maximum isometric muscle strength available (SA 103%, SD 120%). For the hip, the levels of demand were high, but were slightly lower than those of the knee joint. Stair negotiation placed a high level of demand on the knee extensors with demand in SA reaching maximal isometric capacity and demand in SD exceeding maximal isometric capacity. The levels of demand leave little reserve capacity for the older adult to draw on in unexpected situations or circumstances. PMID- 21632254 TI - Unraveling the enigma: progress towards understanding the coronin family of actin regulators. AB - Coronins are a conserved family of actin cytoskeleton regulators that promote cell motility and modulate other actin-dependent processes. Although these proteins have been known for 20 years, substantial progress has been made in the past 5 years towards their understanding. In this review, we examine this progress, place it into the context of what was already known, and pose several questions that remain to be addressed. In particular, we cover the emerging consensus about the role of Type I coronins in coordinating the function of Arp2/3 complex and ADF/cofilin proteins. This coordination plays an important role in leading-edge actin dynamics and overall cell motility. Finally, we discuss the roles played by the more exotic coronins of the Type II and III classes in cellular processes away from the leading edge. PMID- 21632256 TI - Systemic induction of a Capsicum chinense nitrate reductase by the infection with Phytophthora capsici and defence phytohormones. AB - The mRNA differential display technique was used to identify genes from Habanero pepper (Capsicum chinense Jacq.) seedlings whose expression is modified systemically by infection with the oomycete Phytophthora capsici L. Experiments with different oligonucleotide primer combinations revealed that no single gene was synthesised de novo. Instead, the quantitative accumulation of multiple transcripts was found. From these transcripts, levels of a nitrate reductase (Capsicum chinense nitrate reductase, CcNR), which has a high percentage of identity with other Solanaceae NRs, showed a consistent increase a few hours after inoculation (hai) with P. capsici. Reverse northern blotting revealed the existence of basal levels of CcNR transcripts in different adult tissues; however, systemic levels rose dramatically after spraying seedlings with salicylic acid (SA) and ethephon (ET) but not with methyl jasmonate (MeJa). Both P. capsici and defence phytohormones (DP) also modified NR enzymatic activity (nitrite:NAD(+) oxidoreductase; EC 1.7.1.1) with similar kinetics. Because the application of DP induced and activated the CcNR differentially, it is possible that the activity of CcNR is related to a specific host defence response. PMID- 21632257 TI - Haemothorax after mediastinoscopy: a word of caution. AB - Mediastinoscopy is used for the staging of lung cancer and for the diagnosis of mediastinal lesions. It is a valuable diagnostic tool but, in a few cases, it could result in major complications. We describe a simple technique to avoid the development of a major complication--massive haemothorax--after a mediastinoscopy, which should be applied in cases of inadvertent injury of the pleura during the procedure. PMID- 21632258 TI - Long-term results after aortic valve-sparing operation (David I). AB - OBJECTIVE: Aortic valve-sparing David procedure has gained broad acceptance. However, few long-term results have been published. We present our results. METHODS: More than 450 David procedures have been performed in our institution so far. Of these, 126 patients were operated between July 1993 and December 2000. Median age was 57 (8-83) years and 46 (36.5%) were female. As many as 26 (20.6%) had Marfan syndrome, 21 (16.7%) had acute aortic dissection type A (AADA) and 67 (53.2%) had additional procedures. RESULTS: There were six (4.8%) deaths in 30 post-operative period (POD), four of whom had AADA. In the follow-up, there were 32 (25.4%) late deaths, 11 (34.4%) of these were caused by cardiac or underlying disease or op-related. As many as 15 (11.9%) patients were re-operated; six (40%) were Marfan patients and two (13.3%) had early endocarditis. Follow-up echocardiography of 76 (60.3%) event-free patients showed valve insufficiency (AI)<=AI I degrees in 68 (89.5%) and grade II in 7 (9.2%) patients. Leaflet degeneration due to proposed leaflet contact with the straight Dacron graft was not observed. A total of 36 (47.4%) patients were in New York Heart Association (NYHA) class I, 33 (43.4%) in NYHA II, and five (6.6%) were in class III. During the entire follow-up of 790 patient-years, there was no stroke or major bleeding. Survival at 1, 5 and 10 years was 93%, 85% and 70%, respectively. Freedom from valve replacement at 1, 5 and 10 years was 96%, 91% and 87%, respectively. CONCLUSIONS: Regardless of the underlying pathology, valve-sparing David I procedure has acceptable long-term results. Valve-related complications such as stroke or major bleeding is exceedingly low. PMID- 21632260 TI - Is there a morning-to-evening difference in the acute IL-6 and cortisol responses to resistance exercise? AB - Exercise training is known to induce a molecular adaptation process involving inflammatory responses. However any time-of-day effect of exercise on inflammatory responses remains unknown. The aim of the present study was to investigate whether acute bouts of intense exercise performed at different times of the day would affect the release Interleukin-6 (IL-6), one of the most abundant cytokines in mammalian endocrine response to exercise. Cortisol levels were measured as a confirmation of correct timing of exercise and to determine any impact it may have on the cytokine release. Twelve healthy male participants carried out 30 min of intense exercise (3 sets of 8-12 repetitions for 4 resistance exercises at 70% of 1RM) in morning (08:15-09:00 h), and evening (18:15-19:00 h) sessions. An 8h fasting period was required before each exercise session. Blood samples were taken immediately pre and post each exercise sessions to determine IL-6 and cortisol levels. Our data show that whilst the training group showed no post-exercise changes in serum_IL-6 levels (P>0.05), the control group on the other hand showed significant time-of-day modifications in serum_IL 6 levels (P=0.008). Moreover, a significant interaction between intervention phase (pre-post training, AM vs. PM) and group (Exercise vs. Control) is evidenced in terms of serum_IL-6 levels (P=0.014). This interaction however was nullified when the between group differences at baseline were partialled out in a covariate analysis (P>0.05). We also found that the main effect of experimental phase on Cortisol was present in both the trained (P=0.004) and control groups (p<0.001) with no significant interaction (P>0.05). Based on the current data, we would propose that exercise and/or time-of-day would not interfere with clinical endocrine profiling of IL-6 in a population. PMID- 21632259 TI - Oxidative tissue: perilipin 5 links storage with the furnace. AB - Cellular energy homeostasis is a crucial function of oxidative tissues and is altered in obesity, a continuously rising health problem. Lipid droplets (LD) are thought to play a central role in lipid homeostasis by mediating the transient storage of fatty acids in the form of triglyceride, while preventing high levels of toxic lipid intermediates or oxidized lipids that mediate cellular lipotoxicity. Members of the perilipin protein family coating LD surfaces have been found to serve important regulatory and structural functions crucial to the regulation of lipid stores. This review examines the results of studies on one of the newest members of the perilipin family, perilipin 5, which has emerged as a putative key player in LD function in oxidative tissues. PMID- 21632261 TI - A rapid and efficient way to obtain modified chemokines for functional and biophysical studies. AB - Chemokines and their receptors control cell migration associated with routine immune surveillance, inflammation and development. They are also implicated in a large number of inflammatory diseases, cancer and HIV. Here we describe a rapid and efficient way to express and purify milligram quantities of multiple chemokine ligands (CCL7/MCP-3, CCL14/HCC-1, CCL3/MIP-1alpha and CXCL8/IL-8) containing C-terminal modifications to enable coupling to fluorescent dyes or small molecules such as biotin, in vitro. These labeled chemokines display wild type behavior in both receptor binding and calcium mobilization assays. The ability to rapidly and inexpensively produce labeled chemokines opens the way for their use in many applications, including non-traditional chemokine-receptor interaction studies, both on intact cells and with purified receptor reconstituted in artificial membranes in vitro. Furthermore, the ability to immobilize chemokines to obtain ligand affinity columns aids in efforts to purify chemokine receptors for structural and biophysical studies, by facilitating the separation of functional proteins from their non-functional counterparts. PMID- 21632262 TI - TMS effects on subjective and objective measures of vision: stimulation intensity and pre- versus post-stimulus masking. AB - Transcranial magnetic stimulation (TMS) can be used to mask visual stimuli, disrupting visual task performance or preventing visual awareness. While TMS masking studies generally fix stimulation intensity, we hypothesized that varying the intensity of TMS pulses in a masking paradigm might inform several ongoing debates concerning TMS disruption of vision as measured subjectively versus objectively, and pre-stimulus (forward) versus post-stimulus (backward) TMS masking. We here show that both pre-stimulus TMS pulses and post-stimulus TMS pulses could strongly mask visual stimuli. We found no dissociations between TMS effects on the subjective and objective measures of vision for any masking window or intensity, ruling out the option that TMS intensity levels determine whether dissociations between subjective and objective vision are obtained. For the post stimulus time window particularly, we suggest that these data provide new constraints for (e.g. recurrent) models of vision and visual awareness. Finally, our data are in line with the idea that pre-stimulus masking operates differently from conventional post-stimulus masking. PMID- 21632263 TI - Effects of nicotinic acid on endothelial cells and platelets. AB - BACKGROUND: Interactions between platelets and endothelial cells under inflammatory conditions lead to an increased expression of various activity markers of atherosclerosis in the vessel wall. The purpose of this study was to investigate possible protective effects of nicotinic acid in an in vitro endothelial cell model. METHODS: After a 24-hour incubation period with nicotinic acid (1 mmol/l), human umbilical vein endothelial cells were stimulated for 1 h with lipopolysaccharide and were then incubated in direct contact with activated platelets. Following this incubation, the expression of CD40L and CD62P on platelets and the expression of intercellular adhesion molecule (ICAM)-1, vascular cell adhesion molecule (VCAM)-1, uPAR, and MT1-MMP on endothelial cells were measured by flow cytometry. Supernatants were analyzed by ELISA for soluble MCP-1 and MMP-1. RESULTS: The increased expression of VCAM-1 on endothelial cells by proinflammatory stimulation with activated platelets was significantly reduced through preincubation with nicotinic acid (P<.05). Furthermore, platelets in direct contact with preincubated endothelial cells showed a significant reduction in their CD62P and CD40L expression when compared to platelets incubated with untreated endothelial cells (P<.05). Treatment with nicotinic acid did not have a significant effect on ICAM-1, uPAR, and MT1-MMP expression on endothelial cells. Levels of soluble MCP-1 and MMP-1 in supernatants were lower after preincubation with nicotinic acid. CONCLUSION: Nicotinic acid inhibits platelet activation after platelets contacted nicotinic acid treated endothelial cells and inhibits VCAM-1 expression on human endothelial cells under inflammatory conditions. These findings suggest a possible pleiotropic therapeutic relevance of nicotinic acid in atherosclerosis. PMID- 21632264 TI - Limb salvage using bypass to the perigeniculate arteries. AB - OBJECTIVE: To describe bypass to perigeniculate vessels for limb salvage. DESIGN: Retrospective cohort study. MATERIAL AND METHODS: Between 1995 and 2009, 47 bypass procedures to perigeniculate collateral arteries were performed in 46 patients (15 women, 31 men; median age, 68 years). All patients presented with critical ischaemia (tissue loss in 87.5%, rest pain in 12.5%). Mean ankle brachial index was 0.27 +/- 0.17. The site of distal anastomosis was the descending genicular artery (DGA) in 23 bypasses (1 bilateral) and the medial sural artery (MSA) in 24. Proximal anastomosis was to the external iliac artery in 2 cases, common femoral artery in 23 cases, superficial femoral artery in 8 cases, deep femoral artery in 8 cases, above-knee poplitaeal artery in 2 cases, and previous graft in 4 cases. RESULTS: There were four deaths during the immediate postoperative period. Mean follow-up duration was 27 months. Ten patients required major amputation. Mean ankle brachial index post-operatively was 0.60 +/- 0.21. At 3 years, primary patency was 74.7 +/- 7%, secondary patency was 83.4 +/- 8%, and the limb salvage and survival rates were 73.5 +/- 7% and 77.4 +/- 7%, respectively. CONCLUSION: Bypass to perigeniculate arteries is a viable treatment option for critical limb ischaemia in selected patients. PMID- 21632265 TI - A randomised study of perioperative esmolol infusion for haemodynamic stability during major vascular surgery; rationale and design of DECREASE-XIII. AB - OBJECTIVES: This article describes the rationale and design of the DECREASE-XIII trial, which aims to evaluate the potential of esmolol infusion, an ultra-short acting beta-blocker, during surgery as an add-on to chronic low-dose beta-blocker therapy to maintain perioperative haemodynamic stability during major vascular surgery. DESIGN: A double-blind, placebo-controlled, randomised trial. MATERIALS & METHODS: A total of 260 vascular surgery patients will be randomised to esmolol or placebo as an add-on to standard medical care, including chronic low-dose beta blockers. Esmolol is titrated to maintain a heart rate within a target window of 60-80 beats per minute for 24 h from the induction of anaesthesia. Heart rate and ischaemia are assessed by continuous 12-lead electrocardiographic monitoring for 72 h, starting 1 day prior to surgery. The primary outcome measure is duration of heart rate outside the target window during infusion of the study drug. Secondary outcome measures will be the efficacy parameters of occurrence of cardiac ischaemia, troponin T release, myocardial infarction and cardiac death within 30 days after surgery and safety parameters such as the occurrence of stroke and hypotension. CONCLUSIONS: This study will provide data on the efficacy of esmolol titration in chronic beta-blocker users for tight heart-rate control and reduction of ischaemia in patients undergoing vascular surgery as well as data on safety parameters. PMID- 21632267 TI - An initial MRI picture of limbic encephalitis in subacute sclerosing panencephalitis. AB - BACKGROUND: Subacute sclerosing panencephalitis (SSPE) is a rare and severe long term complication of measles. Hallmarks of this entity include progressive cognitive decline, myoclonia, a generalized periodic pattern on EEG and deep white matter abnormalities on MRI. However, imaging can be normal in early stages. AIM: We report herein the case of a previously healthy 13-years-old girl with an unusual radiological presentation. RESULTS: She presented with unilateral myoclonia, cognitive decline with memory impairment and a first brain MRI with swelling of both hippocampi mimicking limbic encephalitis. Measles antibodies were positive in CSF and the EEG showed slow periodic complexes. CONCLUSION: This unusual radiological presentation has never been described in SSPE. Relationship between virus and limbic system are discussed. PMID- 21632268 TI - Exogenous glucocorticoids and adverse cerebral effects in children. AB - Glucocorticoids are commonly used in treatment of paediatric diseases, but evidence of associated adverse cerebral effects is accumulating. The various pharmacokinetic profiles of the exogenous glucocorticoids and the changes in pharmacodynamics during childhood, result in different exposure of nervous tissue to exogenous glucocorticoids. Glucocorticoids activate two types of intracellular receptors, the mineralocorticoid receptor and the glucocorticoid receptor. The two receptors differ in cerebral distribution, affinity and effects. Exogenous glucocorticoids favor activation of the glucocorticoid receptor, which is associated with unfavorable cellular outcomes. Prenatal treatment with glucocorticoids can compromise brain growth and is associated with periventricular leukomalacia, attentions deficits and poorer cognitive performance. In the neonatal period exposure to glucocorticoids reduces neurogenesis and cerebral volume, impairs memory and increases the incidence of cerebral palsy. Cerebral effects of glucocorticoids in later childhood have been less thoroughly studied, but apparent brain atrophy, reduced size of limbic structures and neuropsychiatric symptoms have been reported. Glucocortioids affect several cellular structures and functions, which may explain the observed adverse effects. Glucocorticoids can impair neuronal glucose uptake, decrease excitability, cause atrophy of dendrites, compromise development of myelin producing oligodendrocytes and disturb important cellular structures involved in axonal transport, long-term potentiation and neuronal plasticity. Significant maturation of the brain continues throughout childhood and we hypothesize that exposure to exogenous glucocorticoids during preschool and school age causes adverse cerebral effects. It is our opinion that studies of associations between exposure to glucocorticoids during childhood and impaired neurodevelopment are highly relevant. PMID- 21632269 TI - Non-PKU mild hyperphenylalaninemia (MHP)--the dilemma. AB - Recent reviews have suggested that some patients with "non-PKU mild hyperphenylalaninemia" (MHP) might display neuropsychological executive function deficits and should be considered for treatment with tetrahydrobipterin (BH4) and/or phenylalanine (Phe) restricted diet. Patients with phenylketonuria (PKU)- Classical and Mild/Atypical variants--appear to need "mean lifetime phenylalanine (Phe) levels" of 120-360 MUmol/L for optimal results. MHP patients, on the other hand, have natural Phe levels of 200-600 MUmol/L. Until recently this was thought to be a benign condition. The available literature has been reviewed in detail and no good evidence, to date, has been uncovered to support treatment of MHP. It is suggested that more MHP subjects be tested to confirm this. A plea is made to formulate a consistent world-wide classification of the PKU phenotypes. PMID- 21632271 TI - Lrrk2 p.Q1111H substitution and Parkinson's disease in Latin America. AB - Mutations in the LRRK2 gene are the most common genetic cause of Parkinson's disease, with frequencies displaying a high degree of population-specificity. Although more than 100 coding substitutions have been identified, only seven have been proven to be highly penetrant pathogenic mutations. Studies however are lacking in non-white populations. Recently, Lrrk2 p.Q1111H (rs78365431) was identified in two affected Hispanic brothers and absent in 386 non-Hispanic white healthy controls. We therefore screened this variant in 1460 individuals (1150 PD patients and 310 healthy controls) from 4 Latin American countries (Peru, Chile, Uruguay and Argentina). In our case-control series from Peru and Chile we observed an increased frequency of Lrrk2 p.Q1111H in patients (7.9%) compared to controls (5.4%) although the difference did not reach significance (OR 1.38; p = 0.10). In addition, the frequency of Lrrk2 p.Q1111H varied greatly between populations and further screening in a set of pure Amerindian and pure Spanish controls suggested that this variant likely originated in an Amerindian population. Further studies in other Latin American populations are warranted to assess its role as a risk factor for Parkinson's disease. Screening in Parkinson's disease patients from under-represented populations will increase our understanding of the role of LRRK2 variants in disease risk worldwide. PMID- 21632272 TI - Non-pharmacological and non-surgical interventions for tremor: a systematic review. AB - Tremor is a frequent disabling consequence of many neurological conditions. We performed a search of MEDLINE, CINAHL, EMBASE and the Cochrane Library to identify all primary research studies published up to November 2010 which investigated non-pharmacological and non-surgical treatments for tremor in humans. Neuromuscular physiotherapy, strength training and functional electrical stimulation show promise in their applicability and adaptability. Limb cooling may not be feasible for continuous management, but may be appropriate for specific tasks. Tremor-suppressing orthoses based on viscous materials, weighted splints and vibration therapies need further evaluation especially in the domiciliary setting and applied to improving personal activities of daily living. The evidence base for many rehabilitation interventions in tremor is poor. Future research should focus on high quality randomized controlled trials of non pharmacological and non-surgical interventions which show promise. PMID- 21632273 TI - How to explain central sensitization to patients with 'unexplained' chronic musculoskeletal pain: practice guidelines. AB - Central sensitization provides an evidence-based explanation for many cases of 'unexplained' chronic musculoskeletal pain. Prior to commencing rehabilitation in such cases, it is crucial to change maladaptive illness perceptions, to alter maladaptive pain cognitions and to reconceptualise pain. This can be accomplished by patient education about central sensitization and its role in chronic pain, a strategy known as pain physiology education. Pain physiology education is indicated when: 1) the clinical picture is characterized and dominated by central sensitization; and 2) maladaptive illness perceptions are present. Both are prerequisites for commencing pain physiology education. Face-to-face sessions of pain physiology education, in conjunction with written educational material, are effective for changing pain cognitions and improving health status in patients with various chronic musculoskeletal pain disorders. These include patients with chronic low back pain, chronic whiplash, fibromyalgia and chronic fatigue syndrome. After biopsychosocial assessment pain physiology education comprises of a first face-to-face session explaining basic pain physiology and contrasting acute nociception versus chronic pain (Session 1). Written information about pain physiology should be provided as homework in between session 1 and 2. The second session can be used to correct misunderstandings, and to facilitate the transition from knowledge to adaptive pain coping during daily life. Pain physiology education is a continuous process initiated during the educational sessions and continued within both the active treatment and during the longer term rehabilitation program. PMID- 21632274 TI - Entropy and Laplacian images: structural representations for multi-modal registration. AB - The standard approach to multi-modal registration is to apply sophisticated similarity metrics such as mutual information. The disadvantage of these metrics, in comparison to measuring the intensity difference with, e.g. L1 or L2 distance, is the increase in computational complexity and consequently the increase in runtime of the registration. An alternative approach, which has not yet gained much attention in the literature, is to find image representations, so called structural representations, that allow for the application of the L1 and L2 distance for multi-modal images. This has not only the advantage of a faster similarity calculation but enables also the application of more sophisticated optimization strategies. In this article, we theoretically analyze the requirements for structural representations. Further, we introduce two approaches to create such representations, which are based on the calculation of patch entropy and manifold learning, respectively. While the application of entropy has practical advantages in terms of computational complexity, the usage of manifold learning has theoretical advantages, by presenting an optimal approximation to one of the theoretical requirements. We perform experiments on multiple datasets for rigid, deformable, and groupwise registration with good results with respect to both, runtime and quality of alignment. PMID- 21632275 TI - High-speed atomic force microscopy: Structure and dynamics of single proteins. AB - For surface analysis of biological molecules, atomic force microscopy (AFM) is an appealing technique combining data acquisition under physiological conditions, for example buffer solution, room temperature and ambient pressure, and high resolution. However, a key feature of life, dynamics, could not be assessed until recently because of the slowness of conventional AFM setups. Thus, for observing bio-molecular processes, the gain of image acquisition speed signifies a key progress. Here, we review the development and recent achievements using high speed atomic force microscopy (HS-AFM). The HS-AFM is now the only technique to assess structure and dynamics of single molecules, revealing molecular motor action and diffusion dynamics. From this imaging data, watching molecules at work, novel and direct insights could be gained concerning the structure, dynamics and function relationship at the single bio-molecule level. PMID- 21632276 TI - Phylogeny and evolution of the Archaea: one hundred genomes later. AB - Little more than 30 years since the discovery of the Archaea, over one hundred archaeal genome sequences are now publicly available, of which ~40% have been released in the last two years. Their analysis provides an increasingly complex picture of archaeal phylogeny and evolution with the proposal of new major phyla, such as the Thaumarchaeota, and important information on the evolution of key central cellular features such as cell division. Insights have been gained into the events and processes in archaeal evolution, with a number of additional and unexpected links to the Eukaryotes revealed. Taken together, these results predict that many more surprises will be found as new archaeal genomes are sequenced. PMID- 21632277 TI - Vibrational spectra of double oxalates of the type M(I)2Cu(C2O4)(2).2H2O (MI=Na+, K+, NH4+). AB - The infrared and Raman spectra of Na2Cu(C2O4)(2).2H2O, K2Cu(C2O4)(2).2H2O and (NH4)2Cu(C2O4)(2).2H2O were recorded and briefly discussed on the basis of their structural peculiarities and by comparison with the vibrational spectra of other metallic oxalates. PMID- 21632278 TI - Spectral, structural elucidation and coordination abilities of Co(II) and Mn(II) coordination entities of 2,6,11,15-tetraoxa-9,17-diaza-1,7,10,16-(1,2) tetrabenzenacyclooctadecaphan-8,17-diene. AB - Designing tactics were tailored and followed by synthetic and formulation methodologies to prepare 2,6,11,15-tetraoxa-9,17-diaza-1,7,10,16-(1,2) tetrabenzenacyclooctadecaphan-8,17-diene. Spectral techniques (MS, infrared, 1H NMR, 13C NMR, electronic and EPR), physiochemical measurements (elemental analysis, molar conductance and magnetic susceptibility), electrochemistry (cyclic voltammetry) and classical mechanics (molecular modeling) were employed for structural elucidation of Co(II) and Mn(II) coordination entities having N2O4 chromophore. Comparative spectral analysis revealed legating nature of N2O4 donor macrocycle and confirmed host/guest connectivity between ligand and metal(s). Mass spectrometry (MS) determined 1:1 stoichiometry in CEs. Further electrochemical study confirmed change in oxidation and reduction patterns of CEs. Inhibiting potential (antifungal screened against Aspergillus flavus) showed enhanced antimicrobial properties of CEs as compared to ligand. Molecular modeling was employed to find out different molecular features along with their stabilization energies. PMID- 21632279 TI - Gold nanoparticles-based colorimetric investigation of triplex formation under weak alkalic pH environment with the aid of Ag+. AB - A novel colorimetric method for investigating triplex formation between oligonucleotide modified Au nanoparticles (AuNPs) under weak alkalic pH environment is developed based upon the specific recognition property of Ag+ with CGC triads. Oligonucleotide 5'-SH-T12-CTTCTTTCCTTTCTTC-3' (oligo-1) is modified on the surface of AuNPs. Upon addition of oligonucleotide 5'-GAAGAAAGGAAAGAAG-3' (oligo-2), triplex formation between oligo-1 modified AuNPs occurred at pH 8.0 with the aid of Ag+, triggers the aggregation of AuNPs, accompany with the solution color change from red to purple. The melting temperature demonstrates a 31 degrees C increase for the triplex DNA compose of 10 T*A?T triads and 6 C*G?C triads upon addition of Ag+, the disassociation constant (Kd) between Ag+ and C*G?C triads is 3.6 MUM. Moreover, triplex formation between AuNPs depending on Ag+ can be used to recognize Ag+ ion with the naked eye, as well as UV-vis absorption spectroscopy. PMID- 21632280 TI - Thermal stability and degradation of chitosan modified by benzophenone. AB - N-(biphenylmethylidenyl) chitosan polymer was prepared, characterized and thermal stability was compared with chitosan. Thermal degradation products of the modified polymer were identified by GC-MS technique. It seems that the mechanism of degradation of the prepared polymer is characterized by formation of low molecular weight radicals, followed by random scission mechanism along the backbond chain. PMID- 21632281 TI - Molecular complexes of ketaconazole and oxatomide with p-chloranil: spectroscopic and spectrofluorimetric studies. AB - The molecular complexes of the donors ketaconazole (KTZ) and oxatomide (OXA) drugs with 2,3,5,6-tetrachloro-1,4-benzoquinone (p-chloranil, p-CHL) have been investigated spectroscopically (UV-vis, FT-IR and 1H NMR) and spectrofluorimetrically in different solvents and temperatures. The stoichiometry of the complexes was found to be 1:1. The data are discussed in terms of formation constant, molar extinction coefficient, oscillator strength, dipole moment, ionization potential, dissociation energy and thermodynamic parameters. The results indicated that the formation of molecular complex is spontaneous and endothermic. The fluorescence quenching studies indicated that the interaction of the donors is spontaneous and the fluorescence quenching increased with an increase in the intensity of complexation with the acceptor. PMID- 21632282 TI - Synthesis and photophysical properties of three (multi)branched planar molecules. AB - Three thiophenevinyl substituted one-, two-, and three-branched truxene pi conjugated compounds TS1, TS2 and TS3 have been prepared using a Heck reaction. Their linear absorption, single- and two-photon excited fluorescence were examined. The three analogues emit blue fluorescence at 420 nm. The number of branches has no influence on the position of the absorption maxima of the charge transfer band and fluorescence emitting maxima. However, the molar extinction coefficients of charge transfer band increase almost linearly with the number of branches. The two-photon absorption cross-section of the octupolar three-branched compound TS3 is several times that of the two-branched compound TS2 and one branched compound TS1. PMID- 21632283 TI - Type-specific detection of high-risk human papillomavirus (HPV) in self-sampled cervicovaginal cells applied to FTA elute cartridge. AB - BACKGROUND: Most procedures for self-sampling of cervical cells are based on liquid-based media for transportation and storage. An alternative is to use a solid support, such as dry filter paper media. OBJECTIVES: To evaluate if self sampling of cervicovaginal fluid using a cytobrush (Viba-brush; Rovers Medical Devices B.V., Oss, The Netherlands) and a solid support such as the Whatman Indicating FTA Elute cartridge (GE Healthcare, United Kingdom) can be used for reliable typing of human papillomavirus (HPV), as compared to cervical samples obtained by a physician using a cytobrush and the indicating FTA Elute Micro card and biopsy analysis. STUDY DESIGN: A total of 50 women with a previous high-risk (HR) HPV positive test were invited to perform self-sampling using the Viba-brush and the FTA cartridge and thereafter a physician obtained a cervical sample using the cytobrush and a FTA card, together with a cervical biopsy for histology and HPV typing. Detection of HR-HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58 and 59 was performed using three multiplex real-time polymerase chain reaction (PCR) assays. RESULT: All samples contained sufficient amounts of genomic DNA and the self-samples yielded on average 3.5 times more DNA than those obtained by the physician. All women that were positive for HR-HPV in the biopsy sample also typed positive both by self-sampling and physician-obtained sampling. For women with a histological diagnosis of cervical intraepithelial neoplasia grades 2-3 (CIN 2-3) all three HPV samples showed 100% concordance. A higher number of women were HPV positive by self-sampling than by physician-obtained sampling or by biopsy analysis. CONCLUSION: The Viba-brush and the FTA cartridge are suitable for self-sampling of vaginal cells and subsequent HR-HPV typing. PMID- 21632284 TI - Simple confirmatory assay for anti-HBc reactivity. PMID- 21632285 TI - Magnetic purification of plasminogen from human plasma by specific lysine affinity. AB - A novel magnetic adsorbent was prepared by covalently binding lysine onto the surface of nanoparticles via a carbodiimide coupling method. The adsorption of plasminogen onto surface-modified magnetic nanoparticles from human plasma was studied in a batch system. Surface modifications of particles were characterized using Fourier-transformed infrared spectra, transmission electron micrography, and ninhydrin assay. The maximum weight ratio of lysine to the superparamagnetic particles was 30 MUmol/mg of particles. Effects of pH and temperature on plasminogen adsorption by the magnetic particles were evaluated. Desorption of plasminogen from the magnetic adsorbent was investigated using aminocaproic acid, a lysine analogue. Using a buffer composed by aminocaproic acid or lysine, plasminogen on the magnetic nanoparticles could be eluted. Overall, the results demonstrated that the lysine-coated magnetic adsorbent increased the efficiency and speed of recovery of plasminogen from human plasma. PMID- 21632286 TI - Cytology, liquid-based cytology and automation. AB - Quality-assured, comprehensive, cytology-based population screening programmes have resulted in a substantial decline in the incidence of, and mortality from, cervical cancer. Cytology classification systems divide squamous cell abnormalities into low grade and high grade. Women with high-grade squamous abnormalities, cytology suggestive of glandular neoplasia, or invasive disease are referred immediately for investigation. Previously, the optimal management of women with low-grade cytology was uncertain. The introduction of liquid-based cytology has improved specimen adequacy and laboratory productivity, and also provided the platform for human papillomavirus testing for triage of low-grade abnormality, follow up after treatment and, ultimately, primary screening with triage to cytology, particularly in HPV-vaccinated populations. Liquid-based cytology is also ideal for automation-assisted reading of cervical cytology samples; however, recent studies have reported that automation-assisted reading is less sensitive than manual reading and does not reduce the risk of cervical cancer. PMID- 21632287 TI - Gamma radiation interacts with melanin to alter its oxidation-reduction potential and results in electric current production. AB - The presence of melanin pigments in organisms is implicated in radioprotection and in some cases, enhanced growth in the presence of high levels of ionizing radiation. An understanding of this phenomenon will be useful in the design of radioprotective materials. However, the protective mechanism of microbial melanin in ionizing radiation fields has not yet been elucidated. Here we demonstrate through the electrochemical techniques of chronoamperometry, chronopotentiometry and cyclic voltammetry that microbial melanin is continuously oxidized in the presence of gamma radiation. Our findings establish that ionizing radiation interacts with melanin to alter its oxidation-reduction potential. Sustained oxidation resulted in electric current production and was most pronounced in the presence of a reductant, which extended the redox cycling capacity of melanin. This work is the first to establish that gamma radiation alters the oxidation reduction behavior of melanin, resulting in electric current production. The significance of the work is that it provides the first step in understanding the initial interactions between melanin and ionizing radiation taking place and offers some insight for production of biomimetic radioprotective materials. PMID- 21632288 TI - Efficacy and safety of PANCREAZE(r) for treatment of exocrine pancreatic insufficiency due to cystic fibrosis. AB - BACKGROUND: Pancreatic enzyme replacement therapy (PERT) is critical for correction of exocrine pancreatic insufficiency (EPI) in patients with cystic fibrosis (CF). METHODS: This was a randomized, placebo-controlled PERT withdrawal study evaluating the efficacy and safety of PANCREAZE(r) (pancrelipase) in CF patients with EPI. Participants (n=49) entered an open-label, <= 14 day run-in phase, maintained a high-fat diet (100 +/- 15 g/day), and received PANCREAZE(r) (10.5 or 21). Participants with a coefficient of fat absorption (CFA)>= 80% (n=40) were then randomized (1:1) to receive either PANCREAZE(r) or placebo during a double-blind, <= 7 day withdrawal phase. RESULTS: PANCREAZE(r) improved fat absorption as shown by significantly lower mean +/- SD change in CFA between open-label and double-blind phases for PANCREAZE(r) (-1.5 +/- 5.88%; p<0.001) compared to placebo (-34.1 +/- 23.03%). Protein absorption was similarly improved. No unexpected adverse events were reported. CONCLUSIONS: This study demonstrated PANCREAZE(r) was effective in treating EPI due to CF and was safe and well tolerated. PMID- 21632289 TI - The alternative NADH dehydrogenase is present in mitochondria of some animal taxa. AB - The distribution of the alternative NADH dehydrogenase (NDH-2) in the living world was explored. The enzyme, although present in representatives of all living kingdoms, does not have a universal distribution. With the exception of epsilon proteobacteria, the enzyme was found in all eubacterial groups. In contrast with the known presence of the NDH-2 in Archaea, the alternative oxidase (AOX) is absent in this group. With regard to the Eukarya domain, the NDH-2 was found in representatives of Protista, Fungi, Plantae, and Animalia. In the latter, however, the presence of the enzyme was restricted to some primitive Metazoa (Placozoa and Cnidaria), and two members of the Deuterostomate lineage of the Bilateria (Echinodermata and Urochordata). No evidence for the presence of the NDH-2 was found in any representative of the Protostomate branch of the Bilateria, contrasting with the existence of the AOX in this same group. It is worth mentioning that those animal species containing the NDH-2 also have an AOX. The actual distribution of the NDH-2 in the various living kingdoms is discussed within the framework of the endosymbiotic theory; in addition, a hypothesis is proposed to explain the disappearance of the alternative NDH-2 and AOX from the majority of the animals. PMID- 21632290 TI - The effect of surgical denervation on prevention of excessive dermal scarring: a study on rabbit ear hypertrophic scar model. AB - BACKGROUND: Previous reports have suggested that the extent of wound contraction, epithelisation and total healing time were influenced by denervation of tissues. In this article, we studied for the first time the effect of sensory denervation on prevention of excessive dermal scarring. MATERIALS AND METHODS: Sixteen New Zealand white rabbits were used. Denervation of the right ears was performed by surgical excision of two main sensory nerves. Dissections were also performed on left ears without any nerve excision for the control group. After 14 days of follow-up and confirmation of tissue denervation, an excessive dermal scarring model as defined by Morris et al. was made by surgery on both ears. Twenty-eight days after making the wounds, the tissues were extirpated for analyses. The scars were evaluated by the scar elevation index (SEI), epithelisation time and inflammatory cell count. RESULTS: The SEI of the denervated side scars was significantly lower than that of the non-denervated side. The rate and timing of total epithelisation and inflammatory cell count between groups yielded no difference. CONCLUSIONS: In this study, the surgical denervation skin reduced scarring. It was suggested that understanding the exact role of sensory nerves and neural mediators in excessive dermal scarring is necessary for the prevention and treatment of scarring. PMID- 21632291 TI - Multiscale modeling of calcium dynamics in ventricular myocytes with realistic transverse tubules. AB - Spatial-temporal Ca(2+) dynamics due to Ca(2+) release, buffering, and reuptaking plays a central role in studying excitation-contraction (E-C) coupling in both normal and diseased cardiac myocytes. In this paper, we employ two numerical methods, namely, the meshless method and the finite element method, to model such Ca(2+) behaviors by solving a nonlinear system of reaction-diffusion partial differential equations at two scales. In particular, a subcellular model containing several realistic transverse tubules (or t-tubules) is investigated and assumed to reside at different locations relative to the cell membrane. To this end, the Ca(2+) concentration calculated from the whole-cell modeling is adopted as part of the boundary constraint in the subcellular model. The preliminary simulations show that Ca(2+) concentration changes in ventricular myocytes are mainly influenced by calcium release from t-tubules. PMID- 21632292 TI - Induction of electric field in human bodies moving near MRI: an efficient BEM computational procedure. AB - A computational procedure, based on the boundary element method, has been developed in order to evaluate the electric field induced in a body that moves in the static field around an MRI system. A general approach enables us to investigate rigid translational and rotational movements with any change of motion velocity. The accuracy of the computations is validated by comparison with analytical solutions for simple shaped geometries. Some examples of application of the proposed procedure in the case of motion around an MRI scanner are finally presented. PMID- 21632293 TI - Computed basis functions for finite element analysis based on tomographic data. AB - In bioelectromagnetics, the structures in which the electromagnetic field is to be computed are sometimes defined by a fine grid of voxels (3-D cells) whose tissue types are obtained by tomography. A novel finite element method is proposed for such cases. A simple, regular mesh of cube elements is constructed, each containing the same, integer number of voxels. There may be several different tissues present within an element, but this is accommodated by computing element basis functions that approximately respect the interface conditions between different tissues. Results are presented for a test model of 128 (3) voxels, consisting of nested dielectric cubes, driven by specified charges. The electrostatic potential computed with the new method agrees well with that of a conventional finite element code: the rms difference along the sample line is 1.5% of the highest voltage. Results are also presented for the potential due to a current dipole placed in a brain model of 181 * 217 * 181 voxels, derived from MRI data. The new method gives potentials that are different to those obtained by treating each voxel as an element by 1% of the peak voltage, yet the global finite element matrix has a dimension which is more than 50 times smaller. PMID- 21632294 TI - Tracking colliding cells in vivo microscopy. AB - Leukocyte motion represents an important component in the innate immune response to infection. Intravital microscopy is a powerful tool as it enables in vivo imaging of leukocyte motion. Under inflammatory conditions, leukocytes may exhibit various motion behaviors, such as flowing, rolling, and adhering. With many leukocytes moving at a wide range of speeds, collisions occur. These collisions result in abrupt changes in the motion and appearance of leukocytes. Manual analysis is tedious, error prone,time consuming, and could introduce technician-related bias. Automatic tracking is also challenging due to the noise inherent in in vivo images and abrupt changes in motion and appearance due to collision. This paper presents a method to automatically track multiple cells undergoing collisions by modeling the appearance and motion for each collision state and testing collision hypotheses of possible transitions between states. The tracking results are demonstrated using in vivo intravital microscopy image sequences.We demonstrate that 1)71% of colliding cells are correctly tracked; (2) the improvement of the proposed method is enhanced when the duration of collision increases; and (3) given good detection results, the proposed method can correctly track 88% of colliding cells. The method minimizes the tracking failures under collisions and, therefore, allows more robust analysis in the study of leukocyte behaviors responding to inflammatory conditions. PMID- 21632296 TI - Modelling prostate motion for data fusion during image-guided interventions. AB - There is growing clinical demand for image registration techniques that allow multimodal data fusion for accurate targeting of needle biopsy and ablative prostate cancer treatments. However, during procedures where transrectal ultrasound (TRUS) guidance is used, substantial gland deformation can occur due to TRUS probe pressure. In this paper, the ability of a statistical shape/motion model, trained using finite element simulations, to predict and compensate for this source of motion is investigated. Three-dimensional ultrasound images acquired on five patient prostates, before and after TRUS-probe-induced deformation, were registered using a nonrigid, surface-based method, and the accuracy of different deformation models compared. Registration using a statistical motion model was found to outperform alternative elastic deformation methods in terms of accuracy and robustness, and required substantially fewer target surface points to achieve a successful registration. The mean final target registration error (based on anatomical landmarks) using this method was 1.8 mm. We conclude that a statistical model of prostate deformation provides an accurate, rapid and robust means of predicting prostate deformation from sparse surface data, and is therefore well-suited to a number of interventional applications where there is a need for deformation compensation. PMID- 21632295 TI - Evaluation of registration methods on thoracic CT: the EMPIRE10 challenge. AB - EMPIRE10 (Evaluation of Methods for Pulmonary Image REgistration 2010) is a public platform for fair and meaningful comparison of registration algorithms which are applied to a database of intrapatient thoracic CT image pairs. Evaluation of nonrigid registration techniques is a nontrivial task. This is compounded by the fact that researchers typically test only on their own data, which varies widely. For this reason, reliable assessment and comparison of different registration algorithms has been virtually impossible in the past. In this work we present the results of the launch phase of EMPIRE10, which comprised the comprehensive evaluation and comparison of 20 individual algorithms from leading academic and industrial research groups. All algorithms are applied to the same set of 30 thoracic CT pairs. Algorithm settings and parameters are chosen by researchers expert in the configuration of their own method and the evaluation is independent, using the same criteria for all participants. All results are published on the EMPIRE10 website (http://empire10.isi.uu.nl). The challenge remains ongoing and open to new participants. Full results from 24 algorithms have been published at the time of writing. This paper details the organization of the challenge, the data and evaluation methods and the outcome of the initial launch with 20 algorithms. The gain in knowledge and future work are discussed. PMID- 21632297 TI - Unmixing dynamic fluorescence diffuse optical tomography images with independent component analysis. AB - Dynamic fluorescence diffuse optical tomography (D-FDOT) is important for drug delivery research. However, the low spatial resolution of FDOT and the complex kinetics of drug limit the ability of D-FDOT in resolving metabolic processes of drug throughout whole body of small animals. In this paper, we propose an independent component analysis (ICA)-based method to perform D-FDOT studies. When applied to D-FDOT images, ICA not only generates a set of independent components (ICs) which can illustrate functional structures with different kinetic behaviors, but also provides a set of associated time courses (TCs) which can represent normalized time courses of drug in corresponding functional structures. Further, the drug concentration in specific functional structure at different time points can be recovered by an inverse ICA transformation. To evaluate the performance of the proposed algorithm in the study of drug kinetics at whole-body level, simulation study and phantom experiment are both performed on a full-angle FDOT imaging system with line-shaped excitation pattern. In simulation study, the nanoparticle delivery of indocynaine green (ICG) throughout whole body of a digital mouse is simulated and imaged. In phantom experiment, four tubes containing different ICG concentrations are imaged and used to imitate the uptake and excretion of ICG in organs. The results suggest that we can not only illustrate ICG distributions in different functional structures, but also recover ICG concentrations in specific functional structure at different time points, when ICA is applied to D-FDOT images. PMID- 21632298 TI - A sequential learning algorithm for complex-valued self-regulating resource allocation network-CSRAN. AB - This paper presents a sequential learning algorithm for a complex-valued resource allocation network with a self-regulating scheme, referred to as complex-valued self-regulating resource allocation network (CSRAN). The self-regulating scheme in CSRAN decides what to learn, when to learn, and how to learn based on the information present in the training samples. CSRAN is a complex-valued radial basis function network with a sech activation function in the hidden layer. The network parameters are updated using a complex-valued extended Kalman filter algorithm. CSRAN starts with no hidden neuron and builds up an appropriate number of hidden neurons, resulting in a compact structure. Performance of the CSRAN is evaluated using a synthetic complex-valued function approximation problem, two real-world applications consisting of a complex quadrature amplitude modulation channel equalization, and an adaptive beam-forming problem. Since complex-valued neural networks are good decision makers, the decision-making ability of the CSRAN is compared with other complex-valued classifiers and the best performing real-valued classifier using two benchmark unbalanced classification problems from UCI machine learning repository. The approximation and classification results show that the CSRAN outperforms other existing complex-valued learning algorithms available in the literature. PMID- 21632299 TI - Topology preserving warping of 3-D binary images according to continuous one-to one mappings. AB - The estimation of one-to-one mappings is one of the most intensively studied topics in the research field of nonrigid registration. Although the computation of such mappings can be now accurately and efficiently performed, the solutions for using them in the context of binary image deformation is much less satisfactory. In particular, warping a binary image with such transformations may alter its discrete topological properties if common resampling strategies are considered. In order to deal with this issue, this paper proposes a method for warping such images according to continuous and bijective mappings while preserving their discrete topological properties (i.e., their homotopy type). Results obtained in the context of the atlas-based segmentation of complex anatomical structures highlight the advantages of the proposed approach. PMID- 21632300 TI - Assemble new object detector with few examples. AB - Learning a satisfactory object detector generally requires sufficient training data to cover the most variations of the object. In this paper, we show that the performance of object detector is severely degraded when training examples are limited. We propose an approach to handle this issue by exploring a set of pretrained auxiliary detectors for other categories. By mining the global and local relationships between the target object category and auxiliary objects, a robust detector can be learned with very few training examples. We adopt the deformable part model proposed by Felzenszwalb and simultaneously explore the root and part filters in the auxiliary object detectors under the guidance of the few training examples from the target object category. An iterative solution is introduced for such a process. The extensive experiments on the PASCAL VOC 2007 challenge data set show the encouraging performance of the new detector assembled from those related auxiliary detectors. PMID- 21632301 TI - On dependent bit allocation for multiview image coding with depth-image-based rendering. AB - The encoding of both texture and depth maps of multiview images, captured by a set of spatially correlated cameras, is important for any 3-D visual communication system based on depth-image-based rendering (DIBR). In this paper, we address the problem of efficient bit allocation among texture and depth maps of multiview images. More specifically, suppose we are given a coding tool to encode texture and depth maps at the encoder and a view-synthesis tool to construct intermediate views at the decoder using neighboring encoded texture and depth maps. Our goal is to determine how to best select captured views for encoding and distribute available bits among texture and depth maps of selected coded views, such that the visual distortion of desired constructed views is minimized. First, in order to obtain at the encoder a low complexity estimate of the visual quality of a large number of desired synthesized views, we derive a cubic distortion model based on basic DIBR properties, whose parameters are obtained using only a small number of viewpoint samples. Then, we demonstrate that the optimal selection of coded views and quantization levels for corresponding texture and depth maps is equivalent to the shortest path in a specially constructed 3-D trellis. Finally, we show that, using the assumptions of monotonicity in the predictor's quantization level and distance, suboptimal solutions can be efficiently pruned from the feasible space during solution search. Experiments show that our proposed efficient selection of coded views and quantization levels for corresponding texture and depth maps outperforms an alternative scheme using constant quantization levels for all maps (commonly used in video standard implementations) by up to 1.5 dB. Moreover, the complexity of our scheme can be reduced by at least 80% over the full solution search. PMID- 21632302 TI - An augmented Lagrangian method for total variation video restoration. AB - This paper presents a fast algorithm for restoring video sequences. The proposed algorithm, as opposed to existing methods, does not consider video restoration as a sequence of image restoration problems. Rather, it treats a video sequence as a space-time volume and poses a space-time total variation regularization to enhance the smoothness of the solution. The optimization problem is solved by transforming the original unconstrained minimization problem to an equivalent constrained minimization problem. An augmented Lagrangian method is used to handle the constraints, and an alternating direction method is used to iteratively find solutions to the subproblems. The proposed algorithm has a wide range of applications, including video deblurring and denoising, video disparity refinement, and hot-air turbulence effect reduction. PMID- 21632304 TI - Image magnification using interval information. AB - In this paper, a simple and effective image-magnification algorithm based on intervals is proposed. A low-resolution image is magnified to form a high resolution image using a block-expanding method. Our proposed method associates each pixel with an interval obtained by a weighted aggregation of the pixels in its neighborhood. From the interval and with a linear K(alpha) operator, we obtain the magnified image. Experimental results show that our algorithm provides a magnified image with better quality (peak signal-to-noise ratio) than several existing methods. PMID- 21632303 TI - Human object inpainting using manifold learning-based posture sequence estimation. AB - We propose a human object inpainting scheme that divides the process into three steps: 1) human posture synthesis; 2) graphical model construction; and 3) posture sequence estimation. Human posture synthesis is used to enrich the number of postures in the database, after which all the postures are used to build a graphical model that can estimate the motion tendency of an object. We also introduce two constraints to confine the motion continuity property. The first constraint limits the maximum search distance if a trajectory in the graphical model is discontinuous, and the second confines the search direction in order to maintain the tendency of an object's motion. We perform both forward and backward predictions to derive local optimal solutions. Then, to compute an overall best solution, we apply the Markov random field model and take the potential trajectory with the maximum total probability as the final result. The proposed posture sequence estimation model can help identify a set of suitable postures from the posture database to restore damaged/missing postures. It can also make a reconstructed motion sequence look continuous. PMID- 21632305 TI - Interpolation-dependent image downsampling. AB - Traditional methods for image downsampling commit to remove the aliasing artifacts. However, the influences on the quality of the image interpolated from the downsampled one are usually neglected. To tackle this problem, in this paper, we propose an interpolation-dependent image downsampling (IDID), where interpolation is hinged to downsampling. Given an interpolation method, the goal of IDID is to obtain a downsampled image that minimizes the sum of square errors between the input image and the one interpolated from the corresponding downsampled image. Utilizing a least squares algorithm, the solution of IDID is derived as the inverse operator of upsampling. We also devise a content-dependent IDID for the interpolation methods with varying interpolation coefficients. Numerous experimental results demonstrate the viability and efficiency of the proposed IDID. PMID- 21632306 TI - Complex object correspondence construction in two-dimensional animation. AB - Correspondence construction of objects in key frames is the precondition for inbetweening and coloring in 2-D computer-assisted animation production. Since each frame of an animation consists of multiple layers, objects are complex in terms of shape and structure. Therefore, existing shape-matching algorithms specifically designed for simple structures such as a single closed contour cannot perform well on objects constructed by multiple contours with an open shape. This paper introduces a semisupervised patch alignment framework for complex object correspondence construction. In particular, the new framework constructs local patches for each point on an object and aligns these patches in a new feature space, in which correspondences between objects can be detected by the subsequent clustering. For local patch construction, pairwise constraints, which indicate the corresponding points (must link) or unfitting points (cannot link), are introduced by users to improve the performance of correspondence construction. This kind of input is convenient for animation software users via user-friendly interfaces. A dozen of experimental results on our cartoon data set that is built on industrial production suggest the effectiveness of the proposed framework for constructing correspondences of complex objects. As an extension of our framework, additional shape retrieval experiments on MPEG-7 data set show that its performance is comparable with that of a prominent algorithm published in T-PAMI 2009. PMID- 21632307 TI - Incremental state aggregation for value function estimation in reinforcement learning. AB - In reinforcement learning, large state and action spaces make the estimation of value functions impractical, so a value function is often represented as a linear combination of basis functions whose linear coefficients constitute parameters to be estimated. However, preparing basis functions requires a certain amount of prior knowledge and is, in general, a difficult task. To overcome this difficulty, an adaptive basis function construction technique has been proposed by Keller recently, but it requires excessive computational cost. We propose an efficient approach to this difficulty, in which the problem of approximating the value function is decomposed into a number of subproblems, each of which can be solved with small computational cost. Computer experiments show that the CPU time needed by our method is much smaller than that by the existing method. PMID- 21632308 TI - A wearable, wireless gaze tracker with integrated selection command source for human-computer interaction. AB - A light-weight, wearable, wireless gaze tracker with integrated selection command source for human-computer interaction is introduced. The prototype system combines head-mounted, video-based gaze tracking with capacitive facial movement detection that enable multimodal interaction by gaze pointing and making selections with facial gestures. The system is targeted mainly to disabled people with limited mobility over their hands. The hardware was made wireless to remove the need to take off the device when moving away from the computer, and to allow future use in more mobile contexts. The algorithms responsible for determining the eye and head orientations to map gaze direction to on-screen coordinates are presented together with the one to detect movements from the measured capacitance signal. Point-and-click experiments were conducted to assess the performance of the multimodal system. The results show decent performance in laboratory and office conditions. The overall point-and-click accuracy in the multimodal experiments is comparable to the errors in previous research on head-mounted, single modality gaze tracking that does not compensate for changes in head orientation. PMID- 21632309 TI - The Neurochip-2: an autonomous head-fixed computer for recording and stimulating in freely behaving monkeys. AB - The Neurochip-2 is a second generation, battery-powered device for neural recording and stimulating that is small enough to be carried in a chamber on a monkey's head. It has three recording channels, with user-adjustable gains, filters, and sampling rates, that can be optimized for recording single unit activity, local field potentials, electrocorticography, electromyography, arm acceleration, etc. Recorded data are stored on a removable, flash memory card. The Neurochip-2 also has three separate stimulation channels. Two "programmable system-on-chips" (PSoCs) control the data acquisition and stimulus output. The PSoCs permit flexible real-time processing of the recorded data, such as digital filtering and time-amplitude window discrimination. The PSoCs can be programmed to deliver stimulation contingent on neural events or deliver preprogrammed stimuli. Access pins to the microcontroller are also available to connect external devices, such as accelerometers. The Neurochip-2 can record and stimulate autonomously for up to several days in freely behaving monkeys, enabling a wide range of novel neurophysiological and neuroengineering experiments. PMID- 21632310 TI - Estimated reduction in cancer risk due to PAH exposures if source control measures during the 2008 Beijing Olympics were sustained. AB - BACKGROUND: The 2008 Beijing Olympic Games provided a unique case study to investigate the effect of source control measures on the reduction in air pollution, and associated inhalation cancer risk, in a Chinese megacity. OBJECTIVES: We measured 17 carcinogenic polycyclic aromatic hydrocarbons (PAHs) and estimated the lifetime excess inhalation cancer risk during different periods of the Beijing Olympic Games, to assess the effectiveness of source control measures in reducing PAH-induced inhalation cancer risks. METHODS: PAH concentrations were measured in samples of particulate matter <= 2.5 MUm in aerodynamic diameter (PM2.5) collected during the Beijing Olympic Games, and the associated inhalation cancer risks were estimated using a point-estimate approach based on relative potency factors. RESULTS: We estimated the number of lifetime excess cancer cases due to exposure to the 17 carcinogenic PAHs [12 priority pollutant PAHs and five high-molecular-weight (302 Da) PAHs (MW 302 PAHs)] to range from 6.5 to 518 per million people for the source control period concentrations and from 12.2 to 964 per million people for the nonsource control period concentrations.This would correspond to a 46% reduction in estimated inhalation cancer risk due to source control measures, if these measures were sustained over time. Benzo[b]fluoranthene, dibenz[a,h]anthracene, benzo[a]pyrene, and dibenzo[a,l]pyrene were the most carcinogenic PAH species evaluated. Total excess inhalation cancer risk would be underestimated by 23% if we did not include the five MW 302 PAHs in the risk calculation. CONCLUSIONS: Source control measures, such as those imposed during the 2008 Beijing Olympics, can significantly reduce the inhalation cancer risk associated with PAH exposure in Chinese megacities similar to Beijing. MW 302 PAHs are a significant contributor to the estimated overall inhalation cancer risk. PMID- 21632311 TI - Organization of the root apical meristem in angiosperms. AB - Although flowers, leaves, and stems of the angiosperms have understandably received more attention than roots, the growing root tips, or root apical meristems (RAMs), are organs that could provide insight into angiosperm evolution. We studied RAM organization across a broad spectrum of angiosperms (45 orders and 132 families of basal angiosperms, monocots, and eudicots) to characterize angiosperm RAMs and cortex development related to RAMs. Types of RAM organization in root tips of flowering plants include open RAMs without boundaries between some tissues in the growing tip and closed RAMs with distinct boundaries between apical regions. Epidermis origin is associated with the cortex in some basal angiosperms and monocots and with the lateral rootcap in eudicots and other basal angiosperms. In most angiosperm RAMs, initials for the central region of the rootcap, or columella, are distinct from the lateral rootcap and its initials. Slightly more angiosperm families have exclusively closed RAMs than exclusively open RAMs, but many families have representatives with both open and closed RAMs. Root tips with open RAMs are generally found in angiosperm families considered sister to other families; certain open RAMs may be ancestral in angiosperms. PMID- 21632312 TI - Patterns and development of floral asymmetry in Senna (Leguminosae, Cassiinae). AB - The buzz-pollinated genus Senna (Leguminosae) is outstanding for including species with monosymmetric flowers and species with diverse asymmetric, enantiomorphic (enantiostylous) flowers. To recognize patterns of homology, we dissected the floral symmetry character complex and explored corolla morphology in 60 Senna species and studied floral development of four enantiomorphic species. The asymmetry morph of a flower is correlated with the direction of spiral calyx aestivation. We recognized five patterns of floral asymmetry, resulting from different combinations of six structural elements: deflection of the carpel, deflection of the median abaxial stamen, deflection or modification in size of one lateral abaxial stamen, and modification in shape and size of one or both lower petals. Prominent corolla asymmetry begins in the earl-stage bud (unequal development of lower petals). Androecium asymmetry begins either in the midstage bud (unequal development of thecae in median abaxial stamen; twisting of androecium) or at anthesis (stamen deflection). Gynoecium asymmetry begins in early bud (primordium off the median plane, ventral slit laterally oriented) or midstage to late bud (carpel deflection). In enantiostylous flowers, pronouncedly concave and robust petals of both monosymmetric and asymmetric corollas likely function to ricochet and direct pollen flow during buzz pollination. Occurrence of particular combinations of structural elements of floral symmetry in the subclades is shown. PMID- 21632313 TI - Effects of cell number and cell size on petiole length variation in a stoloniferous herb. AB - In stoloniferous species, the length of petioles is of pivotal importance because it determines the position of leaf blades within the canopy. From a mechanistic perspective, two developmental processes, cell division and cell elongation, are responsible for the length of a given petiole. This study aimed at quantifying the relative contributions of cell division and cell elongation to genotypic and plastic variation in petiole length of the stoloniferous herb Trifolium repens. Thirty-four genotypes of T. repens were grown under high light conditions and simulated canopy shade. Cells were counted and their lengths measured on epidermal prints from fully grown petioles of leaves that had been initiated in the experimental light conditions. Cell number was the main trait explaining petiole length differences among genotypes grown under high light, while both cell number and length changed in response to shading. Our study revealed a strong negative correlation between shade-induced changes in cell number and cell length: genotypes that responded to shading by increasing cell numbers hardly changed in cell length, and vice versa. Our results suggest that genotypic and phenotypic variation in petiole length results from a complex interplay between the developmental processes of cell elongation and cell division. PMID- 21632314 TI - Origins, distribution, and local co-occurrence of polyploid cytotypes in Solidago altissima (Asteraceae). AB - There is growing realization that intraspecific polyploidy in plants has important implications for the evolution of plant diversity and for plant and plant-herbivore community dynamics. Nonetheless, the rate at which polyploid cytotypes arise and their fate in local mixed-cytotype populations are not well understood. Although within- and especially among-population cytotype variation has been documented in many plants, particularly at the largest (continental) spatial scales, local and regional distributions of cytotypes have been well characterized only for a handful of species. Furthermore, few if any studies have addressed both local and regional patterns in the same plant species with sufficient power to detect cytotype variation on both scales. We assessed the co occurrence and frequency distributions of diploid, tetraploid, and hexaploid cytotypes of Solidago altissima (Asteraceae) at local and regional spatial scales, sampling across a zone of presumed ploidy-form overlap in the midwestern United States. Determining ploidy by flow cytometry, we found many local populations containing two or three cytotypes, with the relative frequencies of cytotypes varying dramatically even among neighboring populations. AFLP marker data suggest that polyploid cytotypes of S. altissima likely have multiple origins from different diploid lineages. Our results are consistent with recent perspectives on polyploidization as an evolutionarily dynamic phenomenon with frequent multiple origins of higher-ploidy cytotypes. PMID- 21632315 TI - Population differentiation for plasticity to light in an annual herb: Adaptation and cost. AB - Phenotypic plasticity allows plants to cope with environmental heterogeneity. Environmental variation among populations may select for differentiation in plasticity. To test this idea, we used the annual plant Geranium carolinianum, which inhabits old fields that are densely vegetated and lack canopy cover and wood margins with tree shade but less neighbor shade. Individuals from three populations of each habitat were planted in natural low and high light environments, and morphological traits important for light acquisition were measured. Old-field plants were more plastic, with greater elongation of petioles and internodes in low light than those from wood margins. This larger shade avoidance response suggests evolution of greater plasticity to neighbor shade than to the tree canopy. Fitness of old-field plants was high across both light environments, whereas fitness of wood-margin plants was reduced in low light. Selection favored longer internodes in low than high light. Finally, plasticity for internode length was negatively associated with fitness in high light, suggesting a cost of plasticity for this trait. Together these results indicate that shade-avoidance plasticity of petiole and internode length is adaptive. However, greater elongation of internode length may be constrained by the cost of plasticity expressed in high light. The evolution of plasticity appears to reflect a balance between its adaptive nature and its cost to fitness. PMID- 21632316 TI - Ecological genetics and seed transfer guidelines for Pinus albicaulis (Pinaceae). AB - Whitebark pine (Pinus albicaulis Engelm.) has greatly declined throughout its range as a result of introduced disease, fire suppression, and other factors, and climate change is predicted to accelerate this decline. Restoration is needed; however, no information regarding the degree of local adaptation is available to guide these efforts. A seedling common-garden experiment was employed to assess genetic diversity and geographic differentiation (Q(ST)) of whitebark pine for traits involved in growth and adaptation to cold and to determine climatic variables revealing local adaptation. Seedlings from 48 populations were grown for two years and measured for height increment, biomass, root to shoot ratio, date of needle flush, fall and spring cold injury, and survival. Significant variation was observed among populations for most traits. The Q(ST) was low (0.07 0.14) for growth traits and moderate (0.36-0.47) for cold adaptation related traits, but varied by region. Cold adaptation traits were strongly correlated with mean temperature of the coldest month of population origins, while growth traits were generally correlated with growing season length. We recommend that seed transfer for restoration favor seed movement from milder to colder climates to a maximum of 1.9 degrees C in mean annual temperature in the northern portion of the species range, and 1.0 degrees C in the U. S. Rocky Mountains to avoid maladaptation to current conditions yet facilitate adaptation to future climates. PMID- 21632317 TI - Phytolaccaceae infructescence from Cerro del Pueblo Formation, Upper Cretaceous (late Campanian), Coahuila, Mexico. AB - The Upper Cretaceous (late Campanian) Cerro del Pueblo Formation, Coahuila, Mexico, contains a diverse group of angiosperms represented mainly by their reproductive structures. Among these, a new permineralized infructescence is recognized based on its morphological and anatomical characters. It is a multiple infructescence composed of berry fruits with six locules, each containing a single seed with a curved embryo developed from a campylotropous ovule with pendulous placentation; integumentary anatomy is similar to that of Phytolacca spp. (Phytolaccaceae). Though this new plant from Coahuila shares reproductive characters with Phytolacca, the constant number (six) of carpels per fruit and pendulous placentation strongly support the recognition of a new taxon, Coahuilacarpon phytolaccoides Cevallos-Ferriz, Estrada-Ruiz, et Perez-Hernandez (Phytolaccaceae, Caryophyllales). This new record adds to the known plant diversity of low latitude North America (northern Mexico) and demonstrates the long geologic history of the group. PMID- 21632318 TI - Resistance and tolerance to herbivory changes with inbreeding and ontogeny in a wild gourd (Cucurbitaceae). AB - Herbivory is a ubiquitous component of terrestrial communities that reduces plant growth and reproduction. Consequently, a goal of evolutionary ecology is to identify the causes and consequences of variation in herbivory within plant populations. This three-year study examined the effects of inbreeding on the resistance of wild gourd plants (Cucurbita pepo subsp. texana) to herbivory by cucumber beetles and the impact of the timing of herbivory on reproduction. We grew families of inbred and outbred gourds and recorded beetle damage at three developmental stages, incidence of beetle-vectored wilt disease, survival, and reproduction. While total beetle damage significantly depressed flower and fruit production, damage until mid-July did not depress any measure of reproduction, indicating that these gourds are tolerant of moderate levels of herbivory for most of the growing season. However, beetle damage accumulating after mid-July significantly depressed reproduction, indicating that plants have reduced tolerance during peak reproduction. Early damage, however, did increase the probability of contracting a deadly wilt disease that is vectored by the beetles, suggesting that tolerance and resistance are not alternative defense strategies. Inbreeding significantly reduced resistance to herbivory and, independently of beetle damage, reproductive output. Finally, we found additive genetic variation for both resistance and tolerance that varies with ontogeny. PMID- 21632319 TI - High mycorrhizal specificity in a widespread mycoheterotrophic plant, Eulophia zollingeri (Orchidaceae). AB - Because mycoheterotrophic plants fully depend on their mycorrhizal partner for their carbon supply, the major limiting factor for the geographic distribution of these plants may be the presence of their mycorrhizal partner. Although this factor may seem to be a disadvantage for increasing geographic distribution, widespread mycoheterotrophic species nonetheless exist. The mechanism causing the wide distribution of some mycoheterotrophic species is, however, seldom discussed. We identified the mycorrhizal partner of a widespread mycoheterotrophic orchid, Eulophia zollingeri, using 12 individuals from seven populations in Japan, Myanmar, and Taiwan by DNA-based methods. All fungal ITS sequences from the roots closely related to those of Psathyrella candolleana (Coprinaceae) from GenBank accessions and herbarium specimens. These results indicate that E. zollingeri is exclusively associated with the P. candolleana species group. Further, the molecular data support the wide distribution and wide ranging habitat of this fungal partner. Our data provide evidence that a mycoheterotrophic plant can achieve a wide distribution, even though it has a high mycorrhizal specificity, if its fungal partner is widely distributed. PMID- 21632320 TI - Genetic variation and phylogeography of Stauracanthus (Fabaceae, Genisteae) from the Iberian Peninsula and northern Morocco assessed by chloroplast microsatellite (cpSSR) markers. AB - The tribe Genisteae includes genera of great ecological importance in Mediterranean countries because they are dominant elements of many plant communities. Genetic variation and diversification patterns in Stauracanthus (Genisteae) provide information relevant for the study of the processes of diversification in relation to the environmental history of the western Mediterranean. Nineteen populations of S. boivinii and S. genistoides were assessed by 11 chloroplast microsatellite markers, revealing 44 haplotypes. Both species had different haplotypes and contrasting patterns of karyological, morphological, and genetic variation. In the minimum spanning tree of the haplotypes, AMOVA analysis, and nested clade analysis, S. boivinii had high levels of differentiation and restricted gene flow among populations. Allopatric differentiation occurred between the Moroccan and Iberian populations of S. genistoides, although S. genistoides subsp. spectabilis and subsp. vicentinus had high levels of differentiation among populations (F(ST)), whereas S. genistoides subsp. genistoides had a low F(ST). Genetic patterns are discussed in relation to the Messinian salinity crisis (MSC): hard conditions drove plants to refuge habitats along the Atlantic coast and higher altitude areas in the Moroccan mountains (S. genistoides subsp. spectabilis and S. boivinii). After the MSC, S. boivinii underwent polyploidization and expansion, whereas S. genistoides expanded and continued diversifying into S. genistoides subspp. genistoides and vicentinus. PMID- 21632324 TI - Anatomical basis for biophysical differences between Pinus nigra and P. resinosa (Pinaceae) leaves. AB - Differences in the flexibility of Pinus nigra and P. resinosa leaves can be used to discriminate these two similarly looking pine species from one another. When bent along the longitudinal axis, P. resinosa leaves snap, while P. nigra leaves appear flexible. This useful field test has had no known biophysical or anatomical explanation until now. Analysis of the first order mechanics of bending and buckling of the pine needles was used to elucidate any important anatomical differences between these two species that can account for their different biophysical behaviors when bent. Neither the cross section of the total leaf area nor the inner core area between the two species differed significantly. Differences in the pattern of cell wall thickening and lignification of the endodermal layer of the inner core of the leaves best explain the differences in bending behavior. Thus, subtle variation in anatomy can influence the biophysical properties of naturally occurring structures, which in turn could have important implications for the engineering of manufactured objects. PMID- 21632325 TI - Wood anatomy of Rauvolfioideae (Apocynaceae): a search for meaningful non-DNA characters at the tribal level. AB - Wood anatomical studies in the economically important Apocynaceae or dogbane family are fragmentary. This study represents a first attempt to unravel the phylogenetic significance and major evolutionary trends in the wood of the family, using existing and new microscopic wood observations within the large subfamily Rauvolfioideae. On the basis of LM and SEM observations of 91 species representing all 10 currently recognized tribes, we found that most of the tribes are characterized by a unique combination of wood characters, such as vessel grouping, vessel element length, fiber type, frequency of uniseriate rays, and fused multiseriate rays. Climbing rauvolfioid taxa can generally be distinguished from erect species by their wider vessels, tendency to form paratracheal axial parenchyma, presence of tracheids, and occurrence of laticifers in rays. With respect to the entire family, there is a general phylogenetic trend toward shorter vessel elements, a higher proportion of vessels in multiples and more vessels per multiple, higher tracheid abundance, more paratracheal parenchyma, and fewer cells per axial parenchyma strand in the more derived Apocynaceae. Most of these evolutionary trends are likely to be triggered by drier environmental conditions and/or shifts from an erect to a climbing habit. PMID- 21632326 TI - Plant species diversity and composition of wetlands within an upland forest. AB - Though often overlooked, small wetlands in an upland matrix can support diverse plant communities that increase both local and regional species richness. Here we characterize the full range of wetland vegetation within an upland forest landscape and compare the diversity and composition of different wetland plant communities. In an old-growth forest reserve in southern Quebec, Canada, we sampled wet habitats including lakeshores, permanent and seasonal ponds, swamps, glades, and streamsides. We used clustering, indicator species analysis, and nonmetric multidimensional scaling ordination to identify and compare vegetation types. The wetlands contained 280 species of vascular plants, 45% of the reserve's flora, in only 1.1% of its area. Local diversity averaged 24 +/- 0.7 species per 7 m(2), much higher than in the surrounding upland forests. Plant communities sorted into five types, whose strongest indicator species were Osmunda regalis, Glyceria striata, O. cinnamomea, Deparia acrostichoides, and Matteuccia struthiopteris, respectively. Both local species richness and compositional variation among sites differed among the vegetation types. By combining species representative of the region's major wetlands with species from the upland forest matrix, the plant assemblages of these wetlands make disproportionately important contributions to landscape-level diversity. PMID- 21632327 TI - Sex-specific responses to mycorrhiza in a dioecious species. AB - In most studies about dioecious plants, the role of arbuscular mycorrhizae (AM) and the potential sex-specific differences between the plant hosts have been overlooked. Because plant sexes frequently differ in drought tolerance and AM fungal colonization provides higher resistance to drought, we investigated whether the relation of mycorrhizal fungi with either male or female Antennaria dioica plants differs using a factorial experiment. We hypothesized that because AM usually increase growth rate and male plants usually grow larger than females, males should gain more benefit from the mycorrhizal symbiosis in terms of mineral nutrition and water supply. Because of higher demands of carbohydrates (C) in males, we expected males to allocate less C resources to the mycorrhizal fungus so that the associated fungi should benefit less of the association with males. In contrast to our initial hypothesis, the male plants, although faster growing under drought, did not gain more symbiosis-mediated benefits than did the females, and both sexes seemed to provide resources equally to their fungal symbiont. Therefore, we conclude that the two plant sexual morphs provide equal amounts of C to their fungal root symbionts and that they can gain specific benefits from the symbiosis, which, however, depend on soil water availability. PMID- 21632328 TI - Nutrient stress and gall flies interact to affect floral-sex ratio in gynomonoecious Solidago altissima (Asteraceae). AB - A main tenet of sex-allocation theory is that environmental stress should lead to increased maleness because reproducing through pollen is generally cheaper than producing fruits and seeds. Though this prediction has held for many species, it has been little tested for gynomonoecious plants, in which individuals produce both female and perfect flowers. We exposed eight ramets of each of 22 genets of a gynomonoecious goldenrod, Solidago altissima (Asteraceae), to a factorial combination of nutrient stress and herbivory by the gall-inducer Eurosta solidaginis (Tephritidae). Nutrient stress alone increased relative femaleness: Stressed ramets produced fewer flowers total and a higher ratio of ray (female) flowers to disk (perfect) flowers. Galling caused no change in fertilized ramets, but the combination of nutrient stress and galling caused an increase in relative maleness: Nutrient-stressed, galled ramets produced fewer flowers total and had a higher disk to ray ratio. In addition to being phenotypically plastic, floral-sex ratio had a great deal of genetic variation, with a broad-sense heritability of 0.68. While the floral-sex ratio responses of gynomonoecious plants may be more complicated than for plants of other breeding systems, they offer the potential to test and refine the already rich body of sex-allocation theory. PMID- 21632329 TI - Population structure and genetic diversity of New World maize races assessed by DNA microsatellites. AB - Because of the economic importance of maize and its scientific importance as a model system for studies of domestication, its evolutionary history is of general interest. We analyzed the population genetic structure of maize races by genotyping 964 individual plants, representing almost the entire set of ~350 races native to the Americas, with 96 microsatellites. Using Bayesian clustering, we detected four main clusters consisting of highland Mexican, northern United States (US), tropical lowland, and Andean races. Phylogenetic analysis indicated that the southwestern US was an intermediary stepping stone between Mexico and the northern US. Furthermore, southeastern US races appear to be of mixed northern flint and tropical lowland ancestry, while lowland middle South American races are of mixed Andean and tropical lowland ancestry. Several cases of post Columbian movement of races were detected, most notably from the US to South America. Of the four main clusters, the highest genetic diversity occurs in highland Mexican races, while diversity is lowest in the Andes and northern US. Isolation by distance appears to be the main factor underlying the historical diversification of maize. We identify highland Mexico and the Andes as potential sources of genetic diversity underrepresented among elite lines used in maize breeding programs. PMID- 21632330 TI - Comparison of clonal diversity in mountain and Piedmont populations of Trillium cuneatum (Melanthiaceae-Trilliaceae), a forest understory species. AB - The balance between clonal and sexual reproduction can vary widely among plant populations, and the extent of clonality may be influenced by the combined effects of historical land use and variation in environmental conditions. We investigated patterns of clonal spread in five Trillium cuneatum populations, two in the Appalachian Mountains characterized by mesic, cooler conditions, and three at lower elevations experiencing warmer, drier conditions and greater disturbance. Using a new measure of the genet effective number and innovative orthogonal contrast methods, we quantified genet structure, contrasting clonal growth in the mountains with that in the Piedmont. Asexual propagation was more common in the Piedmont, where 25% of the sampled ramets were clonally derived, but was much less frequent in the mountains (7% clonal replicates). Hierarchical partitioning of variation in genet diversity showed that the majority (75.8%) of the variation resulted from more vegetative replication in the Piedmont. Most of the remaining variation (21.6%) was attributable to differences between urban and rural Piedmont populations, and a small, statistically nonsignificant fraction of the variation (2.6%) was due to interpopulation differences within the mountains. Higher frequency of cloning may enhance both genetic and demographic population viability in fragmented Piedmont habitats. PMID- 21632331 TI - Phylogeography of North African Atlas cedar (Cedrus atlantica, Pinaceae): Combined molecular and fossil data reveal a complex Quaternary history. AB - Northwest Africa is a major hotspot of plant biodiversity, but very little is known about the Quaternary range dynamics of plant species in this region. Here we investigate the range-wide population structure and phylogeography of Atlas cedar (Cedrus atlantica), an emblematic forest tree endemic to Morocco and Algeria. We genotyped 261 individuals from 11 populations using AFLP markers. Data were analyzed using both conventional F(ST)-based techniques and Bayesian clustering. Overall population differentiation was high (F(ST) = 0.25). Two major groups of populations were identified, one distributed through the Rif and Middle Atlas mountains in Morocco and the other through the Algerian Tell Atlas and Aures mountains as well as the Middle Atlas. Combined molecular and fossil data indicate that C. atlantica survived the Last Glacial Maximum in at least three disjunct refugia along the coast of the Mediterranean Sea, whereas the Middle Atlas, today the core of the species range, has been colonized relatively recently (<10000 yr BP). The colonization history of individual populations has left clear imprints in their present-day diversity, which may vary greatly even between nearby stands. Our study illustrates how integrating different data sources and analytical approaches can help elucidate complex range dynamics that would otherwise remain undeciphered. PMID- 21632332 TI - Phylogenetics of neotropical Platymiscium (Leguminosae: Dalbergieae): systematics, divergence times, and biogeography inferred from nuclear ribosomal and plastid DNA sequence data. AB - Platymiscium is a neotropical legume genus of forest trees in the Pterocarpus clade of the pantropical "dalbergioid" clade. It comprises 19 species (29 taxa), distributed from Mexico to southern Brazil. This study presents a molecular phylogenetic analysis of Platymiscium and allies inferred from nuclear ribosomal (nrITS) and plastid (trnL, trnL-F and matK) DNA sequence data using parsimony and Bayesian methods. Divergence times are estimated using a Bayesian method assuming a relaxed molecular clock (multidivtime). Within the Pterocarpus clade, new sister relationships are recovered: Pterocarpus + Etaballia, Inocarpus + Tipuana and Paramachaerium + Maraniona. Our results support monophyly of Platymiscium, which is resolved into three major clades, each with distinct geographic ranges and ecological preferences. Diversification in Platymiscium has been driven by habitat fragmentation, invasion of novel geographic regions, and ecological diversification, revealing general patterns of diversification in the neotropics. We hypothesize that Platymiscium arose in dry habitats of South America and radiated northward. The Amazon basin was invaded twice both within the last 5.6 My and Central America twice before the closure of the Isthmus of Panama. Divergence times of the P. pubescens complex, restricted to seasonally dry tropical forests of South America, support pre-Pleistocene divergence in this biome. PMID- 21632333 TI - Genetic diversity and endemism in North American Carex section Ceratocystis (Cyperaceae). AB - Factors leading to endemism, and the evolutionary implications of endemism, can be explored by studying closely related taxa with variously restricted distributions. Such a model is provided by Carex section Ceratocystis (Cyperaceae); Carex cryptolepis, Carex sp. nov., and C. lutea are North American endemics with broad, restricted, and highly restricted distributions, respectively. The prediction that levels of genetic diversity are a consequence of distribution size was tested within a phylogenetic context using population level genetic variation at 18 allozyme loci. In contrast to expectations, mean proportion of loci polymorphic, number of alleles per polymorphic locus, and expected heterozygosity were significantly greater in C. lutea than either C. cryptolepis or Carex sp. nov. Although the possibility of a shift in breeding system, past introgression, or progenitor-derivative relationships could explain the relatively high levels of variation observed in C. lutea, these were dismissed on the basis of allozyme and nuclear ribosomal sequence data. We conclude that C. lutea maintains levels of genetic diversity typical of caespitose carices despite its narrow endemism and that the low levels of genetic variation in C. cryptolepis and Carex sp. nov. are likely the result of population fluctuations during Pleistocene glacial-interglacial cycles. PMID- 21632334 TI - Hybridization, polyploidy, and evolutionary transitions between monoecy and dioecy in Bryonia (Cucurbitaceae). AB - Correns's 1903 (Berichte der Deutschen Botanischen Gesellschaft 21: 133-147) crosses between a monoecious and a dioecious species of Bryonia revealed the simple Mendelian inheritance of dioecy and provided the first instance of an XY sex determination system in any organism. Bryonia ranges from the Canary Islands to Central Asia and comprises seven dioecious and three monoecious species; its closest relative, Ecballium elaterium, has dioecious and monoecious populations. We used chloroplast (cp) and nuclear (nr) gene phylogenies to infer sexual system evolution in Bryonia. We also tested for associations between sexual system and ploidy level, based on published and original chromosome counts. Conflicts between cp and nr topologies imply that the dioecious hexaploid B. cretica arose from hybridization(s), probably involving the dioecious diploids B. dioica, B. syriaca, and/or B. multiflora. A tetraploid dioecious endemic on Corsica and Sardinia probably originated from B. dioica via autopolyploidy. While the cp phylogeny resolves few species relationships, the nr tree implies at least two evolutionary changes in sexual system. There is no correlation between sexual system and ploidy level. Molecular clocks suggest that the deepest divergence, between a species on the Canary Islands and the ancestor of all remaining species, occurred ca. 10 million years ago. PMID- 21632335 TI - Brassicaceae phylogeny inferred from phytochrome A and ndhF sequence data: tribes and trichomes revisited. AB - The family Brassicaceae comprises 3710 species in 338 genera, 25 recently delimited tribes, and three major lineages based on phylogenetic results from the chloroplast gene ndhF. To assess the credibility of the lineages and newly delimited tribes, we sequenced an approximately 1.8-kb region of the nuclear phytochrome A (PHYA) gene for taxa previously sampled for the chloroplast gene ndhF. Using parsimony, likelihood, and Bayesian methods, we reconstructed the phylogeny of the gene and used the approximately unbiased (AU) test to compare phylogenetic results from PHYA with findings from ndhF. We also combined ndhF and PHYA data and used a Bayesian mixed model approach to infer phylogeny. PHYA and combined analyses recovered the same three large lineages as those recovered in ndhF trees, increasing confidence in these lineages. The combined tree confirms the monophyly of most of the recently delimited tribes (only Alysseae, Anchonieae, and Descurainieae are not monophyletic), while 13 of the 23 sampled tribes are monophyletic in PHYA trees. In addition to phylogenetic results, we documented the trichome branching morphology of species across the phylogeny and explored the evolution of different trichome morphologies using the AU test. Our results indicate that dendritic, medifixed, and stellate trichomes likely evolved independently several times in the Brassicaceae. PMID- 21632336 TI - Palaeoanellus dimorphus gen. et sp. nov. (Deuteromycotina): a Cretaceous predatory fungus. AB - In habitats where nitrogen is the limiting factor, carnivorous fungi gain an advantage by preying on nematodes and other microorganisms. These fungi are abundant in modern terrestrial ecosystems, but they are not predestined for preservation as fossils. Conclusions on their evolutionary history are therefore mainly based on molecular studies that are generally limited to those taxa that have survived until today. Here we present a fossil dimorphic fungus that was found in Late Albian amber from southwestern France. This fungus possessed unicellular hyphal rings as trapping devices and formed blastospores from which a yeast stage developed. The fossil probably represents an anamorph of an ascomycete and is described as Palaeoanellus dimorphus gen. et sp. nov. Because predatory fungi with regular yeast stages are not known from modern ecosystems, the fungus is assumed to not be related to any Recent carnivorous fungus and to belong to an extinct lineage of carnivorous fungi. The inclusions represent the only record of fossil fungi that developed trapping devices, so far. The fungus lived c. 100 million years ago in a limnetic-terrestrial microhabitat, and it was a part of a highly diverse biocenosis at the forest floor of a Cretaceous coastal amber forest. PMID- 21632337 TI - Molecular insights into the evolution of crop plants. AB - The domestication and improvement of crop plants have long fascinated evolutionary biologists, geneticists, and anthropologists. In recent years, the development of increasingly powerful molecular and statistical tools has reinvigorated this now fast-paced field of research. In this paper, we provide an overview of how such tools have been applied to the study of crop evolution. We also highlight lessons that have been learned in light of a few long-standing and interrelated hypotheses concerning the origins of crop plants and the nature of the genetic changes underlying their evolution. We conclude by discussing compelling evolutionary genomic approaches that make possible the efficient and unbiased identification of genes controlling crop-related traits and provide further insight into the actual timing of selection on particular genomic regions. PMID- 21632338 TI - Unisexual flower, spikelet, and inflorescence development in monoecious/dioecious Bouteloua dimorpha (Poaceae, Chloridoideae). AB - Unisexual flowers have evolved repeatedly in the angiosperms. In Poaceae, multiple transitions from bisexual to unisexual flowers are hypothesized. There appear to be at least three distinct developmental mechanisms for unisexual flower formation as found in members of three subfamilies (Ehrhartoideae, Panicoideae, Pharoideae). In this study, unisexual flower development is described for the first time in subfamily Chloridoideae, as exemplified by Bouteloua dimorpha. Scanning electron microscopy (SEM) and anatomy were used to characterize the development of male (staminate) and female (pistillate) flowers, spikelets, and inflorescences. We found the developmental pathway for staminate flowers in B. dimorpha to be distinct from that described in the other three subfamilies, showing gynoecial arrest occurs at a different stage with possible loss of some cellular contents. However, pistillate flowers of B. dimorpha had some similarity to those described in other unisexual-flowered grasses, with filament and anther differentiation in abortive stamens. Comparing our findings with previous reports, unisexual flowers seem to have evolved independently in the four examined grass subfamilies. This analysis suggests the action of different genetic mechanisms, which are consistent with previous observations that floral unisexuality is a homoplasious condition in angiosperms. PMID- 21632339 TI - Hormone interactions and regulation of PsPK2::GUS compared with DR5::GUS and PID::GUS in Arabidopsis thaliana. AB - The putative pea PINOID homolog, PsPK2, is expressed in all growing plant parts and is positively regulated by auxin, gibberellin, and cytokinin. Here, we studied hormonal regulation of PsPK2::GUS expression compared with DR5::GUS and PID::GUS in Arabidopsis. PsPK2::GUS, DR5::GUS, and PID::GUS expression in Arabidopsis shoots is mainly localized in the stipules, hydathodes, veins, developing leaves, and cotyledons. Unlike DR5::GUS, PsPK2::GUS, and PID::GUS are weakly expressed in root tips. Both DR5::GUS and PsPK2::GUS are induced by different auxins and are more sensitive to methyl indole acetic acid, 4-chloro indole acetic acid, and alpha-naphthalene acetic acid than others. GA(3) has no significant effect on GUS activity in DR5::GUS-transformed seedlings compared to the control, but induction by auxin and gibberellin in combination is synergistic. Cytokinin increases auxin transport in Arabidopsis seedlings. Auxin, gibberellin, and cytokinin all increase GUS activity in shoots of PsPK2::GUS transformed plants compared to the control. However, only auxin and gibberellin increase GUS activity in PID::GUS shoots. In conclusion, auxin, gibberellin, and cytokinin positively regulate PsPK2 expression in shoots, but not in roots. Auxin and gibberellin also upregulate AtPIN1 and LEAFY expression, which is similar to PsPIN1 and Uni in pea. With minor exceptions, the orthologous genes from both species are regulated similarly. PMID- 21632340 TI - The relative fitness of parental and hybrid Kunzea (Myrtaceae): The interaction of reproductive traits and ecological selection. AB - Up to 22% of plant species are the result of breeding among species-hybridization directly conflicting with the prediction that hybrids, compared to parental species, are intermediate in character and of low fitness and little consequence. Few studies, however, have compared the fitness of hybrids and parental species under field conditions. This study evaluates components of fitness in the field for naturally occurring hybrids of the shrub Kunzea, relative to the parental speciesKunzea rupestris. Hybrid plants did not differ from the parental species in the level of effective pollination. Thus, we found no support for Grant's model (Evolution: International Journal of Organic Evolution3: 82-97) of reduced fitness of hybrids via reduced pollination level (the intermediate hypothesis). Hybrids displayed variable fitness across the measured fitness components. Seed set levels for hybrids were structured among populations, suggesting genetic structuring for this fitness component at this scale. The response of hybrids to fire (a major selective force in the study system) was partly consistent with a resource trade-off model. Hybrids were large robust plants but most did not resprout after fire. Hence, the fitness of hybrids was complex. We developed a model for relative fitness to estimate fitness for species and hybrids with complex life histories. PMID- 21632341 TI - Mycorrhizal interactions of orchids colonizing Estonian mine tailings hills. AB - Northeastern Estonia is home to extensive oil shale mines. Associated with these are desolate and environmentally damaging hills of ash and semicoke tailings. Interestingly, some of the first plants to colonize these hills are rare orchids. Here, we assess the identities of the mycorrhizal fungi associated with these orchids, in particular Epipactis atrorubens, Orchis militaris, and Dactylorhiza baltica, and compare them with mycorrhizal fungi from orchids from pristine habitat. Epipactis atrorubens associated with the widest breadth of fungi, including unnamed members of the basidiomycete family Tulasnellaceae and the potentially ectomycorrhizal ascomycetes Trichophaea woolhopeia and Geopora cooperi. Orchis militaris also associated with unnamed members of the Tulasnellaceae. Dactylorhiza baltica associated with Ceratobasidium albasitensis. In Epipactis and Orchis, the same fungi associated with plants in the pristine habitat as with those on ash hills. The tulasnelloid and ceratobasidioid fungi mycorrhizal with these orchids appear closely related to common orchid mycorrhizal fungi, while one of the ascomycetes mycorrhizal with E. atrorubens is closely related to a mycorrhizal fungus with E. microphylla. Our results suggest that these orchids and their fungi are not limited to pristine habitats and that environmentally polluted sites may present novel habitats that may be exploited for endangered plant conservation. PMID- 21632342 TI - Effects of elevated CO2 on the tolerance of photosynthesis to acute heat stress in C3, C4, and CAM species. AB - Determining the effect of elevated CO(2) on the tolerance of photosynthesis to acute heat stress (AHS) is necessary for predicting plant responses to global warming because photosynthesis is heat sensitive and AHS and atmospheric CO(2) will increase in the future. Few studies have examined this effect, and past results were variable, which may be related to methodological variation among studies. In this study, we grew 11 species that included cool and warm season and C(3), C(4), and CAM species at current or elevated (370 or 700 ppm) CO(2) and at species-specific optimal growth temperatures and at 30 degrees C (if optimal ? 30 degrees C). We then assessed thermotolerance of net photosynthesis (P(n)), stomatal conductance (g(st)), leaf internal [CO(2)], and photosystem II (PSII) and post-PSII electron transport during AHS. Thermotolerance of P(n) in elevated (vs. ambient) CO(2) increased in C(3), but decreased in C(4) (especially) and CAM (high growth temperature only), species. In contrast, elevated CO(2) decreased electron transport in 10 of 11 species. High CO(2) decreased g(st) in five of nine species, but stomatal limitations to P(n) increased during AHS in only two cool-season C(3) species. Thus, benefits of elevated CO(2) to photosynthesis at normal temperatures may be partly offset by negative effects during AHS, especially for C(4) species, so effects of elevated CO(2) on acute heat tolerance may contribute to future changes in plant productivity, distribution, and diversity. PMID- 21632343 TI - Plastid movement in statocytes of the arg1 (altered response to gravity) mutant. AB - The ability of a plant to respond to gravity is crucial for growth and development throughout the life cycle. A key player in the cellular mechanisms of gravitropism is ARG1 (altered response to gravity), a DnaJ-like protein that associates with components of the vesicular trafficking pathway and carries a C terminal domain with similarities to cytoskeleton-associated proteins. The arg1-2 mutant of Arabidopsis thaliana has reduced and delayed gravitropism in roots, shoots, and inflorescence stems when grown in the light or dark. We performed light microscopic studies of plastid movement in the gravity-perceiving statocytes (endodermal cells) of hypocotyls of arg1-2 and WT light-grown seedlings following reorientation to better characterize the role of ARG1 in gravitropism. Cryofixation/freeze substitution procedures were used because they provide a reliable indication of rapid cellular events within the statocytes. Our results suggest that ARG1 affects gravitropism by reducing plastid movement/sedimentation, a process known to be essential for early phases of signaling cascades in the statocytes. PMID- 21632344 TI - Geographic variation in floral morphology and style-morph ratios in a sexually polymorphic daffodil. AB - Narcissus triandrus is a tristylous daffodil from the Iberian Peninsula that has striking geographical variation in floral morphology and style-morph ratios. Here, we investigate the relation between this variation and ecological factors to understand mechanisms governing morph ratios. We estimated morph ratios in 124 populations throughout the range of N. triandrus and measured 13 morphological traits in 35-78 populations. Sampling of morph ratios conducted over 2-10-yr intervals demonstrated strong temporal stability. Variation in floral and vegetative traits enabled statistical prediction of morph-frequency variation among populations. Latitudinal gradients in precipitation and temperature were correlated with plant and flower size, with larger flowers in northern populations associated with bumblebee visitation and stylar dimorphism. Flowers of the L- and M-morphs differed significantly from the S-morph in several size related characters, unlike other tristylous species. This pattern and the similarity in anther positions of the L- and M-morphs suggest that the M-morph of N. triandrus originated through genetic modifiers that shortened styles of the L morph. Our findings support the hypothesis that geographical variation in style morph ratios is largely governed by climatic gradients in the Iberian Peninsula, which influence the floral morphology and pollination biology of N. triandrus populations. PMID- 21632345 TI - Morphology and biogeography of Apiaceae subfamily Saniculoideae as inferred by phylogenetic analysis of molecular data. AB - The phylogenetic placements of several African endemic genera at the base of Apiaceae subfamilies Saniculoideae and Apioideae have revolutionized ideas of relationships that affect hypotheses of character evolution and biogeography. Using an explicit phylogeny of subfamily Saniculoideae, we reconstructed the evolutionary history of phenotypic characters traditionally important in classification, identified those characters most useful in supporting relationships, and inferred historical biogeography. The 23 characters examined include those of life history, vegetative morphology, inflorescences, and fruit morphology and anatomy. These characters were optimized over trees derived from maximum parsimony analysis of chloroplast DNA trnQ-trnK sequences from 94 accessions of Apiaceae. The results revealed that many of these characters have undergone considerable modification and that traditional assumptions regarding character-state polarity are often incorrect. Infrasubfamilial relationships inferred by molecular data are supported by one to five morphological characters. However, none of these morphological characters support the monophyly of subfamilies Saniculoideae or Apioideae, the clade of Petagnaea, Eryngium and Sanicula, or the sister-group relationship between Eryngium and Sanicula . Southern African origins of Saniculoideae and of its tribes Steganotaenieae and Saniculeae are supported based on dispersal-vicariance analysis. PMID- 21632346 TI - Molecular phylogenetics of the species-rich neotropical genus Swartzia (Leguminosae, Papilionoideae) and related genera of the swartzioid clade. AB - Swartzia (ca. 180 spp.) is a characteristic and diverse element of neotropical rainforest tree communities. As such, it has been identified as a focal group in studies of evolutionary diversification and community assembly in rainforests. However, progress is impeded by the lack of a phylogenetic hypothesis for the genus and its close relatives, which together constitute the descendents of one of the first branches of the papilionoid phylogeny. Here we present a molecular phylogenetic study with extensive sampling of species of Swartzia and with complete sampling of genera of the "swartzioid" clade. The results, based on analysis of chloroplast (atpB-rbcL, trnL intron, and trnL-F) and nuclear (AAT1 and ITS) DNA sequences, add substantially to our understanding of relationships within this diverse group and call for taxonomic changes, particularly within Swartzia. The monophyly of a redefined Swartzia is weakly to moderately supported. Within Swartzia, the analyses identify 11 nonoverlapping subclades, few of which correspond exactly to previously published taxa. The analyses support the recent segregations of Bobgunnia from Swartzia and of Trischidium from Bocoa, as well as the resurrection of the monospecific genus Fairchildia. The analyses identify a "core swartzioid" clade comprising the remainder of Swartzia, Bocoa, and Candolleodendron. PMID- 21632347 TI - Evolution of secondary heads in Nassauviinae (Asteraceae, Mutisieae). AB - The evolution of the inflorescence head in Asteraceae is important in the diversification of this largest angiosperm family. The aggregation of heads into higher-order capitulescences (secondary heads or syncephalia) is considered evolutionarily advanced. The genera Moscharia, Nassauvia, Polyachyrus, and Triptilion of the subtribe Nassauviinae (Mutisieae) have syncephalia with differing degrees of capitula condensation. ITS and plastid trnL-trnF regions were analyzed separately and together using maximum parsimony and maximum likelihood to examine the evolution of syncephalia in the Nassauviinae. The four genera displaying syncephalia do not form a clade minus taxa without syncephalia, indicating that secondary heads in Nassauviinae have either convergently evolved twice in the subtribe (or, very unlikely) once with multiple reversions. Strong support was obtained for a sister relationship between Leucheria (without syncephalium) and Polyachyrus, and both sister to Moscharia. Nassauvia and Triptilion form a distinct clade but are sister to other genera, Perezia and Panphalea, without syncephalium. Previous hypotheses postulated the evolution from simple to more complex secondary heads. We show that the ancestor of Moscharia, Polyachyrus, and Leucheria, in a more arid habitat, had a complex type of secondary head, and loss of complexity occurred in response to a shift from arid to mesic conditions. PMID- 21632348 TI - Phylogenetics and reticulate evolution in Pistacia (Anacardiaceae). AB - The systematic position and intrageneric relationships of the economically important Pistacia species (Anacardiaceae) are controversial. The phylogeny of Pistacia was assessed using five data sets: sequences of nuclear ribosomal ITS, the third intron of the nuclear nitrate reductase gene (NIA-i3), and the plastid ndhF, trnL-F and trnC-trnD. Significant discordance was detected among ITS, NIA i3, and the combined plastid DNA data sets. ITS, NIA-i3, and the combined plastid data sets were analyzed separately using Bayesian and parsimony methods. Both the ITS and the NIA-i3 data sets resolved the relationships among Pistacia species well; however, these two data sets had significant discordance. The ITS phylogeny best reflects the evolutionary relationships among Pistacia species. Lineage sorting of the NIA-i3 alleles may explain the conflicts between the NIA-i3 and the ITS data sets. The combined analysis of three plastid DNA data sets resolved Pistacia species into three major clades, within which only a few subclades were supported. Pistacia was shown to be monophyletic in all three analyses. The previous intrageneric classification was largely inconsistent with the molecular data. Some Pistacia species appear not to be genealogical species, and evidence for reticulate evolution is presented. Pistacia saportae was shown to be a hybrid with P. lentiscus (maternal) and P. terebinthus (paternal) as the parental taxa. PMID- 21632349 TI - DNA from herbarium specimens settles a controversy about origins of the European potato. AB - Landrace potato cultivars are native to two areas in South America: the high Andes from eastern Venezuela to northern Argentina and the lowlands of south central Chile. Potato first appeared outside of South America in Europe in 1567 and rapidly diffused worldwide. Two competing hypotheses suggested the origin of the "European" potato from the Andes or from lowland Chile, but the Andean origin has been widely accepted over the last 60 years. All modern potato cultivars predominantly have Chilean germplasm, explained as originating from breeding with Chilean landraces subsequent to the late blight epidemics beginning in 1845 in the UK. The Andean origin has been questioned recently through examination of landraces in India and the Canary Islands, but this evidence is inferential. Through a plastid DNA deletion marker from historical herbarium specimens, we report that the Andean potato predominated in the 1700s, but the Chilean potato was introduced into Europe as early as 1811 and became predominant long before the late blight epidemics in the UK. Our results provide the first direct evidence of these events and change the history of introduction of the European potato. They shed new light on the value of past breeding efforts to recreate the European potato from Andean forms and highlight the value of herbarium specimens in investigating origins of crop plants. PMID- 21632350 TI - Life span correlates with population dynamics in perennial herbaceous plants. AB - Survival and fecundity are basic components of demography and therefore have a strong influence on population dynamics. These two key parameters and their relationship are crucial to understand the evolution of life histories. It remains, however, to be empirically established how life span, fecundity, and population dynamics are linked in different organism groups. We conducted a comparative study based on demographic data sets of 55 populations of 23 perennial herbs for which structured demographic models and among-year natural variation in demographic attributes were available. Life span (from 4 to 128 yr old), estimated by using an algorithm, was inversely correlated with the deviance of the population growth rate from equilibrium as well as with among-year population fluctuations. Temporal variability was greater for short-lived species than for the long-lived ones because fecundity was more variable than survival and relatively more important for population dynamics for the short-lived species. The relationship between life span and population stability suggests that selection for longevity may have played an important role in the life history evolution of plants because of its ability to buffer temporal fluctuations in population size. PMID- 21632351 TI - Growth pattern and age determination for Cecropia sciadophylla (Urticaceae). AB - Cecropia species, ranging from Mexico to northern Argentina and the West Indies, are pioneer trees that colonize cleared areas with high light. To determine their ages to help pinpoint the date of the area's disturbance, we need to understand their developmental and architectural changes over time. The simple architecture of Cecropia conforms to the model of Rauh; that is, it has orthotropic axes with lateral flowering and rhythmic branching. The axes are made of a succession of nodes and internodes whose length and associated lateral productions remain measurable for years. Thus, by describing the tree trunk node by node, we can depict the sequence of events involved in tree development. For 25 trees of C. sciadophylla, from two stations in French Guiana and Colombia, we recorded internode length and any presence of branches, and flowers for each node. Using autocorrelation coefficients, we found a high periodicity in flowering and branching, with inflorescences at every 25 nodes, stages of branches spaced by a multiple of 25 nodes, and alternation of long and short nodes every 25 nodes. Considering that flowering is annual for many Cecropia species, the main conclusion of this work is that C. sciadophylla has strong annual growth, branching, and flowering rhythms. In addition, the age of the tree can be estimated retrospectively by observing its adult morphology. PMID- 21632352 TI - Embryology of Manekia naranjoana (Piperaceae) and the origin of tetrasporic, 16 nucleate female gametophytes in Piperales. AB - The vast majority of flowering plant seeds contain a triploid endosperm formed by fertilization of a monosporic, Polygonum-type female gametophyte. However, evolutionary transitions to six other genetic constructs of endosperm are widespread, and six of seven known patterns are found in the order Piperales. Within Piperaceae, Manekia has not been described, and we report its female gametophyte to be tetrasporic and 16-nucleate at maturity. Manekia ontogeny is generally characterized by early establishment of a bipolar or weakly bipolar body plan and a binucleate central cell at maturity (Drusa-type pattern); however, ca. 16% of early stages had distinctly tetrapolar organization, and ca. 21% of mature specimens had a tetranucleate central cell (Penaea-type pattern, not previously reported in Piperaceae). An evolutionary developmental analysis indicates heterochrony, heterotopy, novelties, and sequence deletions have each played roles in modulating variation within Piperales. Our data suggest the common ancestor of Piperaceae was tetrasporic and retained a plesiomorphic bipolar body plan, producing a "functionally bisporic" form of triploid endosperm derived from the lineal descendants of two megaspores and a sperm. Developmental modifications of this tetrasporic, bipolar ontogeny can account for the origin of all three other known "true" tetrasporic endosperm genetic constructs, formed from derivatives of all four megaspores and a sperm. These derived endosperms in turn have higher ploidy, higher potential heterozygosity, and reduced genetic conflicts. PMID- 21632353 TI - An exploration of LEAFY expression in independent evolutionary origins of rosette flowering in Brassicaceae. AB - Whereas most Brassicaceae produce flowers on an elongated inflorescence, a few lineages produce flowers directly from the vegetative rosette on elongated pedicels. Knowing the extent to which independent origins of rosette flowering involve the same developmental and genetic mechanisms could clarify the constraints acting on plant architectural evolution. Prior work in Idahoa, Ionopsidium, and Leavenworthia suggested that changes in the activity or expression of the flower meristem identity gene, LEAFY (LFY), played a role in all three origins of rosette flowering. Here we studied the developmental morphology of L. crassa and immunolocalization of LFY protein in Leavenworthia and Ionopsidium to further compare independent origins of rosette flowering. Leavenworthia crassa differs from Ionopsidium and Idahoa in producing ebracteate flowers. Flowers are, however, associated with "squamules," here interpreted as stipules of a cryptic bract. LFY was detected in L. crassa flower primordia but not in inflorescence meristems. In contrast, the rosette flowering Io. acaule accumulated LFY protein in the inflorescence meristem, whereas its inflorescence flowering close relative, Io. prolongoi, did not. Thus, although different cases of rosette flowering likely entailed modifications of the same meristem identity program, distinct developmental genetic mechanisms appear to be involved in each case. PMID- 21632354 TI - Density-dependent mating and reproductive assurance in the temperate forest herb Paris quadrifolia (Trilliaceae). AB - In animal-pollinated plants, autonomous selfing provides reproductive assurance under conditions of infrequent pollinator visits or a lack of mates, but few data are available for wind-pollinated species or species with combined insect and wind-pollination, for which it is often assumed that pollen availability does not limit reproduction. In this study, the capacity of autonomous selfing was investigated in the temperate forest herb Paris quadrifolia, and an emasculation experiment was performed under natural field conditions to investigate the contribution of autonomous selfing to total seed set across a continuous gradient of densities of flowering conspecifics. In the absence of wind or pollinators, autonomous selfing was observed through anthers approaching stigmas at the end of flowering and the capacity for autonomous pollination was about 0.34. Under natural conditions, considerable outcross pollination was observed, but the proportion of ovules successfully fertilized significantly decreased with decreasing density of conspecifics when flowers were emasculated, but not when flowers were left intact. These results indicate that autonomous selfing resulted in reproductive assurance (RA = 0.16) and thus support the hypothesis that autonomous selfing can also provide reproductive assurance in wind-pollinated species. PMID- 21632355 TI - Hydraulic traits are influenced by phylogenetic history in the drought-resistant, invasive genus Juniperus (Cupressaceae). AB - In the conifer genus Juniperus (Cupressaceae), many species are increasing rapidly in distribution, abundance, and dominance in arid and semiarid regions. To help understand the success of junipers in drier habitats, we studied hydraulic traits associated with their water stress resistance, including vulnerability to xylem cavitation, specific conductivity (K(S)), tracheid diameter, conduit reinforcement, and wood density in stems and roots, as well as specific leaf area (SLA) of 14 species from the United States and the Caribbean. A new phylogeny based on DNA sequences tested the relationships between vulnerability to cavitation and other traits using both traditional cross-species correlations and independent contrast correlations. All species were moderately to highly resistant to water-stress-induced cavitation in both roots and shoots. We found strong phylogenetic support for two clades previously based on leaf margin serration (serrate and smooth). Species in the serrate clade were 34-39% more resistant to xylem cavitation in stems and roots than were species in the smooth clade and had ~35% lower K(S) and 39% lower SLA. Root and stem resistance to cavitation and SLA were all highly conserved traits. A high degree of conservation within clades suggests that hydraulic traits of Juniperus species strongly reflect phylogenetic history. The high resistance to cavitation observed may help explain the survival of junipers during recent extreme droughts in the southwestern United States and their expansion into arid habitats across the western and central United States. PMID- 21632356 TI - Pollinator foraging modifies nectar sugar composition in Helleborus foetidus (Ranunculaceae):An experimental test. AB - We experimentally tested the hypothesis that the extensive within-plant variation of nectar sugar composition in Helleborus foetidus (Ranunculaceae) and other species results from differences between flowers and nectaries in pollinator visitation history. Experiments were conducted to mimic single-nectary visits by wild-caught individuals of the main bee pollinators of H. foetidus, which were assayed for their capacity to modify the sugar composition of natural and artificial nectar. Experimental nectar probing with bee mouthparts induced extensive changes in proportional sugar composition 48 h after treatment, and bee taxa differed widely in their effects. Nectar probing by Andrena, medium-sized Anthophoridae, Apis mellifera, and Lasioglossum had no subsequent effects on nectar sugar composition, while probing by Bombus terrestris and B. pratorum induced an extensive reduction in percentage sucrose, a marked increase in percentage fructose, and a slight increase in percentage glucose. Results support the hypothesis that stochastic variations among flowers or nectaries in the taxonomic identity of recent visitors and their relative visitation frequencies may eventually generate very small-scale mosaics in nectar sugar composition. Changes in nectar sugar composition following bumblebee probing may be the consequence of nectar contamination with pollinator-borne nectarivorous yeasts. PMID- 21632357 TI - Dispersal limitations, rather than bottlenecks or habitat specificity, can restrict the distribution of rare and endemic rainforest trees. AB - Despite their narrow distribution, Australian rainforests still contain considerable levels of diversity and include many ancient, but often rare, lineages. Very little is known about the general biology of rainforest species, yet their long-term management depends on a better understanding of the main factors leading to rarity. For instance, are they highly endemic taxa, at the early stages of expansion, nearing the end of a period of decline, or persisting at low numbers over the long term? In this study we combine molecular, environmental, and ecological data to identify the factors responsible for the narrow distribution of a paleoendemic rainforest tree: Elaeocarpus sedentarius (Elaeocarpaceae). Between-population and between-generation comparisons of genetic diversity across all known populations of E. sedentarius show evidence of mutation-drift equilibrium rather than evidence of a recent bottleneck. Similarly, floristic and environmental data negate the hypothesis of rarity as a consequence of highly specialized habitat requirements. Instead, genetic structure and the available ecological data support the hypothesis of dispersal limitation as the main cause of endemism and that the species may have attained genetic equilibrium without realizing its full niche potential. We suggest that these factors are likely to explain narrow endemism in a broader range of taxa. PMID- 21632358 TI - Todea from the Lower Cretaceous of western North America: implications for the phylogeny, systematics, and evolution of modern Osmundaceae. AB - The first fossil evidence for the fern genus Todea has been recovered from the Lower Cretaceous of British Columbia, Canada, providing paleontological data to strengthen hypotheses regarding patterns of evolution and phylogeny within Osmundaceae. The fossil consists of a branching rhizome, adventitious roots, and leaf bases. The dictyoxylic stem has up to eight xylem bundles around a sclerenchymatous pith. Leaf traces diverge from cauline bundles in a typical osmundaceous pattern and leaf bases display a sheath of sclerenchyma around a C shaped xylem trace with 2-8 protoxylem strands. Within the adaxial concavity of each leaf trace, a single sclerenchyma bundle becomes C-shaped as it enters the cortex. The sclerotic cortex is heterogeneous with an indistinct outer margin. The discovery of Todea tidwellii sp. nov. reveals that the genus Todea evolved by the Lower Cretaceous. A phylogenetic analysis combining morphological characters of living and extinct species with a previously published nucleotide sequence matrix confirms the taxonomic placement of T. tidwellii. Results also support the hypothesis that Osmunda s.l. represents a paraphyletic assemblage and that living species be segregated into two genera, Osmunda and Osmundastrum. Fossil evidence confirms that Osmundaceae originated in the Southern Hemisphere during the Permian, underwent rapid diversification, and species extended around the world during the Triassic. Crown group Osmundaceae originated by the Late Triassic, with living species appearing by the Late Cretaceous. PMID- 21632359 TI - Scutifolium jordanicum gen. et sp. nov. (Cabombaceae), an aquatic fossil plant from the Lower Cretaceous of Jordan, and the relationships of related leaf fossils to living genera. AB - A new species of aquatic plant, Scutifolium jordanicum gen. et sp. nov., Taylor, Brenner & Basha, is described from the Albian of Jordan. The leaves are microphyllous with a symmetrical, elliptical to suborbiculate shape, convex to rounded apex and base, and actinodromous to palinactinodromous primary venation. The peltate, centrally attached petioles are narrow, elongate, and alternately arranged on similarly sized stems. The leaves appear to be thick and have aerenchyma. Comparisons to plants with centrally peltate leaves and palmate venation and to aquatic plants with floating leaves suggest that S. jordanicum belongs to the Cabombaceae lineage within the Nymphaeales. Cladistic analysis including the fossil and living members of the Nymphaeales shows that the S. jordanicum is basal to the living members of the family and has unique characters not found in any living genera. This is the oldest evidence of the Cabombaceae from the Old World. Inclusion of two other Early Cretaceous peltate leaf fossils in the phylogenetic analysis indicates their affinities to Cabombaceae and that some of the shared derived characters for the living members are progressively acquired in the fossils. These data show the Cabombaceae were widespread in Gondwana and Laurasia by the mid-Cretaceous. PMID- 21632360 TI - AFLP markers as a tool to reconstruct complex relationships: A case study in Rosa (Rosaceae). AB - The genus Rosa has a complex evolutionary history caused by several factors, often in conjunction: extensive hybridization, recent radiation, incomplete lineage sorting, and multiple events of polyploidy. We examined the applicability of AFLP markers for reconstructing (species) relationships in Rosa, using UPGMA clustering, Wagner parsimony, and Bayesian inference. All trees were well resolved, but many of the deeper branches were weakly supported. The cluster analysis showed that the rose cultivars can be separated into a European and an Oriental cluster, each being related to different wild species. The phylogenetic analyses showed that (1) two of the four subgenera (Hulthemia and Platyrhodon) do not deserve subgeneric status; (2) section Carolinae should be merged with sect. Cinnamomeae; (3) subsection Rubigineae is a monophyletic group within sect. Caninae, making sect. Caninae paraphyletic; and (4) there is little support for the distinction of the five other subsections within sect. Caninae. Comparison of the trees with morphological classifications and with previous molecular studies showed that all methods yielded reliable trees. Bayesian inference proved to be a useful alternative to parsimony analysis of AFLP data. Because of their genome wide sampling, AFLPs are the markers of choice to reconstruct (species) relationships in evolutionary complex groups. PMID- 21632361 TI - An AFLP marker approach to lower-level systematics in Eucalyptus (Myrtaceae). AB - Genus Eucalyptus, with over 700 species, presents a number of systematic difficulties including taxa that hybridize or intergrade across environmental gradients. To date, no DNA marker has been found capable of resolving phylogeny below the sectional level in the major subgenera. Molecular markers are needed to support taxonomic revision, assess the extent of genetic divergence at lower taxonomic levels, and inform conservation efforts. We examined the utility of 930 amplified fragment length polymorphisms (AFLPs) for analyzing relationships among Tasmanian taxa of subgenus Symphyomyrtus section Maidenaria. Phenetic and cladistic analyses resolved species into clusters demonstrating significant genetic partitioning, largely concordant with series defined in the most recent taxonomic revision of Eucalyptus. Some departures from current taxonomy were noted, indicating possible cases of morphological convergence and character reversion. Although the resolution obtained using AFLP was greatly superior to that of single sequence markers, the data demonstrated high homoplasy and incomplete resolution of closely related species. The results of this study and others are consistent with recent speciation and reticulate evolution in Maidenaria. We conclude that a combination of phylogenetic and population genetic approaches using multiple molecular markers offers the best prospects for understanding taxonomic relationships below the sectional level in Eucalyptus. PMID- 21632362 TI - Evolution of Synthyris sect. Dissecta (Plantaginaceae) on sky islands in the Northern Rocky Mountains. AB - We examined the phylogeography of Synthyris sect. Dissecta (Plantaginaceae), which is restricted to the Olympic (S. lanuginosa) and Northern Rocky Mountains (S. dissecta and S. canbyi), to infer effects of sky islands and modes of speciation. Sequences of cpDNA trnT-trnL and psbA-trnH intergenic spacers resolved 22 haplotypes among 185 individuals sampled from 16 populations of the three species. Gene flow in the ancestral lineage and random capture of haplotypes in species lineages of sect. Dissecta have resulted in haplotype clades that are not exclusive to species. Nested clade analysis (NCA) indicates that allopatric fragmentation separated Olympic and Northern Rocky Mountain populations, giving rise to the Olympic endemic S. lanuginosa, which is characterized by unique haplotypes consistent with long temporal isolation. Low haplotype and nucleotide diversity in S. canbyi are consistent with newly founded populations experiencing a bottleneck. Furthermore, we infer S. canbyi evolved as a peripheral isolate of S. dissecta. NCA indicated limited migration in S. dissecta with possible isolation by distance. Both isolation on interglacial sky islands and valley glaciers during at least the last glaciation limited gene flow among populations of S. dissecta in different ranges of the Northern Rocky Mountains. PMID- 21632363 TI - Geographic diversity cline of R gene homologs in wild populations of Solanum pimpinellifolium (Solanaceae). AB - Plant resistance (R) genes tend to be highly variable within plant species and are thought to be under natural selection; however, little is known about the geographic distribution of R gene diversity within and among plant populations. To determine the possible roles of demography and selection on R gene evolution, patterns of diversity at the multigenic Cf-2 R gene family were studied in Solanum pimpinellifolium populations along the northern coast of Peru. Population diversity levels of Cf-2 homologs follow a latitudinal cline, consistent with the species's history of gradual colonization of the Peruvian coast and population variation in outcrossing levels. Although previous evidence suggests that selection has shaped the DNA sequence content of the Cf-2 genes, current results imply that the geographic distribution of Cf-2 homolog diversity has been shaped primarily by demographic factors or by selective pressures with a clinal distribution. PMID- 21632364 TI - Pollen and anther ontogeny in Cabomba caroliniana (Cabombaceae, Nymphaeales). AB - Cabomba is a small water lily genus that is native to the New World. Studies of pollen development and associated changes in the anther yield valuable characters for considering the evolution of reproductive biology in seed plants. Here we characterized the complete ontogenetic sequence for pollen in Cabomba caroliniana. Anthers at the microspore mother cell, tetrad, free microspore, and mature pollen grain stages were studied using scanning electron, transmission electron, and light microscopy. Tetragonal and decussate tetrads both occur in C. caroliniana, indicating successive microsporogenesis. The exine is tectate columellate, and the infratectal columellae are the first exine elements to form, followed by a continuous tectum and a thin foot layer. A lamellate endexine initiates in the early free microspore stage, but becomes compressed in mature grains. Tectal microchannels and sculptural rods also initiate during the early free microspore stage, and significant pollenkitt deposition follows, supporting the hypothesis that these elements function in entomophily. The tapetum is morphologically amoeboid, with migratory tapetal cells directly contacting developing free microspores within the anther locule. Results from this study illustrate the importance of including ontogenetic data in analyzing pollen characters and in developing evolutionary and ecological hypotheses. The new palynological data also emphasize the character plasticity that occurs in basal angiosperms. PMID- 21632365 TI - Growth, development, and systematics of ferns: Does Botrychium s.l. (Ophioglossales) really produce secondary xylem?1. AB - Developmental morphology and anatomy of Botrychium s.l. were studied to clarify rhizome ontogeny and patterns of tissue maturation that can be used to test the hypothesis that ferns of the Ophioglossales may represent living progymnosperms. Serial anatomical sections of the rhizomes of B. virginianum and B. dissectum reveal that apical meristematic activity and vascular tissue maturation occur over an extended period of several years and then stop. Most of the xylem consists of radial rows of tracheids and interspersed ray-like xylem parenchyma cells that are similar in these respects to secondary xylem, but pits occur on all tracheid walls as is characteristic of primary xylem. No vascular cambium is initiated in mature primary tissues, nor is there secondary phloem. Radial rows of xylem cells are produced by the direct continuation of divisions that begin at the shoot apical meristem, forming a cylinder of radially aligned procambial cells before the differentiation of protoxylem. Continuing divisions over a period of several years increase the number of thin-walled cells and tracheids in each radial row back to about one internode behind where the current year's frond trace diverges from the rhizome stele. At more proximal levels of the rhizome, procambial cell divisions cease and there is no additional tracheid differentiation. These data reveal that the rhizome matures over an exceedingly long period of several years, but that growth is ultimately determinate, thus supporting hypotheses that the Ophioglossales is more closely related to other groups of living ferns than to progymnosperms and seed plants. PMID- 21632366 TI - Genetic effects on the biomass partitioning and growth of Pisum and Lycopersicon. AB - We examined a series of eight pea genotypes differing in three naturally occurring allelic mutations, i.e., af (afila), st (stipules reduced), and tl (tendril-less) and three species, five cultivars, and one interspecific hybrid of tomato differing in SP (SELF-PRUNING) allele composition to determine whether different phenotypes ontogenetically express different biomass partitioning patterns compared to the isometric partitioning pattern and an interspecific 3/4 scaling "rule" governing annual growth with respect to body mass. The slopes and "elevations" (i.e., alpha and log beta, respectively) of log-log linear regression curves of bivariate plots of leaf, stem, and root dry mass and of annual growth vs. total body mass were used to assess pattern homogeneity. The annual growth of all pea and tomato phenotypes complied with the 3/4 growth rule. The biomass partitioning patterns of all tomato phenotypes were statistically indistinguishable from the isometric pattern as were those of the pea wild type and three single-mutant genotypes. However, significant departures from the isometric (and pea wild type) biomass allocation pattern were observed for three genotypes, all of which were homozygous for the af allele. These results open the door to explore the heritability and genetics underlying the allometry of biomass partitioning patterns and growth. PMID- 21632367 TI - Influence of nutrient availability on the mechanisms of tolerance to herbivory in an annual grass, Avena barbata (Poaceae). AB - Tolerance, or the capacity of a genotype to survive and reproduce following herbivore damage, varies widely across the plant kingdom. One proximate cause of this variation is resource availability, which can influence tolerance through mechanisms such as growth rate and photosynthesis. We examined the effect of high and low soil nutrient levels on the relationship between tolerance and two of its underlying mechanisms, biomass regrowth and photosynthetic upregulation, among genotypes of the Mediterranean annual grass Avena barbata. Although defoliated plants did not reach the same biomass as controls, biomass regrowth was higher at high nutrients. However, increased seed abortion at high nutrients caused tolerance to be the same in both nutrient treatments. Increased seed abortion also uncoupled biomass regrowth from tolerance at high nutrients. We found no evidence for photosynthetic upregulation in defoliated compared to control plants in either nutrient treatment. However, tolerance was positively correlated with predefoliation photosynthetic efficiency at high nutrients. Thus, constitutive photosynthetic efficiency may be a better predictor of tolerance than photosynthetic responses following herbivory in A. barbata. More generally, our results highlight the possibility that the mechanisms of tolerance can differ across resource environments even if tolerance is the same. PMID- 21632368 TI - Stage-dependent border cell and carbon flow from roots to rhizosphere. AB - Rising CO(2) levels in the atmosphere have drawn attention to the important role of soil in sequestering carbon. This project goal was to quantify soil carbon deposition associated with border cell release and exudation from root growth zones. Carbon was measured with a Carlo Erba C/N analyzer in soil from the rhizosphere of mature grasses and, in separate experiments, in soil collected around root growth zones. Root border cells in "rhizosphere soil" (silica sand) were counted using a compound microscope after soil sonication and extraction with surfactant. For sand-grown Bromus carinatus, Zea mays, and Cucumis sativus, young seedlings (with roots shorter than 2 cm) released thousands of border cells, while older root tips released only hundreds. For a variety of native annual and perennial grasses and invasive annual grasses (Nassella pulchra, B. carinatus, B. diandrus, B. hordeaceus, Vulpia microstachys, Aegilops triuncialis, Lolium multiflorum, Zea mays), the rhizosphere of mature root systems contained between 18 and 32 MUg C g(-1) sand more than that of the unplanted controls. Spatial analysis of the rhizosphere around the cucumber growth zone confirmed C enrichment there. The root tip provided C to the rhizosphere: 4.6 MUg C in front of the growing tip, with the largest deposition, 20.4 MUg C, to the rhizosphere surrounding the apical 3 mm (root cap/meristem). These numbers from laboratory studies represent the maximum C that might be released during flooding in soils. Scaling up from the organ scale to the field requires a growth analysis to quantify root tip distributions in space and time. PMID- 21632369 TI - Survival and performance of the invasive vine Vincetoxicum rossicum (Apocynaceae) from seeds of different embryo number under two light environments. AB - The nonnative vine Vincetoxicum rossicum threatens several ecosystems in the Lower Great Lakes Basin of North America. One feature that may contribute to its invasiveness is the production of some seeds with multiple embryos (polyembryony), which may be beneficial as a bet-hedging strategy in variable environments. However, lower seed reserves per embryo in polyembryonic seeds may entail costs in low-light environments. The effect of seed from three embryonic classes (1, 2, or 3 embryos/seed) on V. rossicum survival and growth was studied under two forest understory light environments: full canopy (shade) or canopy gaps (light) in New York state. Two seedling cohorts were planted, in May 2004 and in May 2005. The survival and growth of seedlings was monitored biweekly for two (2005 cohort) or three (2004 cohort) seasons. For both cohorts, plants grown in canopy shade had reduced survival and growth compared with those grown in gaps. Contrary to expectations, seed embryo number had no effect on the final height, survival, or dry mass of plants in either habitat. Our results suggest that any fitness advantage provided by polyembryony may be habitat (light) dependent and not a general trait that affords V. rossicum a benefit in all habitats colonized. PMID- 21632370 TI - Molecular phylogeny and reticulate origins of the polyploid Bromus species from section Genea (Poaceae). AB - The origin of polyploid Bromus species of section Genea was investigated using molecular data. This group of annual species native from the Old-World is composed of three diploids, two tetraploids, one hexaploid, and one octoploid. Molecular cloning, sequencing, and phylogenetic analyses were performed on several accessions per species. We used the low copy nuclear gene Waxy, repeated rDNA spacers ITS1 and ITS2 and chloroplast spacers trnT-trnL and trnL-trnF. Our analyses revealed four different lineages involved in the parentage of the polyploids and confirmed their reticulate origin. Three of these lineages are closely related to the diploid species B. sterilis, B. tectorum, and B. fasciculatus. The fourth lineage could not be related to any diploid according to the available data. Our data gave insights on the origin of all the polyploids of section Genea, and chloroplast data allowed us to identify the maternal lineages. The Waxy gene was the most informative regarding origin of the polyploids. The Waxy copies duplicated by polyploidy appear selectively maintained in the polyploid species. No sequence heterogeneity was encountered in the ITS region, where concerted evolution seems to have occurred toward either maternal or paternal repeats. These results provide new information about the origin and molecular evolution of these polyploids and will allow a more accurate taxonomic treatment of the concerned species, based on their evolutionary history. PMID- 21632371 TI - Seed ferns survived the end-Cretaceous mass extinction in Tasmania. AB - Seed ferns, dominant elements of the vegetation in many parts of the world from the Triassic to Cretaceous, were considered to have disappeared at the end of the Cretaceous together with several other groups that had occupied key positions in terrestrial and marine ecosystems such as dinosaurs, plesiosaurs, and ammonoids. Seed-fern demise is generally correlated with competition from diversifying flowering plants through the Cretaceous and the global environmental crisis related to the Chicxulub impact event in the paleotropics at the end of the period. New fossils from Tasmania show that one seed-fern lineage survived into the Cenozoic by at least 13 million years. These fossils are described here as a new species, Komlopteris cenozoicus. Komlopteris is a genus of seed ferns attributed to Corystospermaceae and until now was not known from sediments younger than the Early Cretaceous. Discovery of this "Lazarus taxon," together with the presence of a range of other relictual fossil and extant organisms in Tasmania, other southern Gondwanan provinces, and some regions of northern North America and Asia, underscores high-latitude regions as biodiversity refugia during global environmental crises and highlights their importance as sources of postextinction radiations. PMID- 21632372 TI - Scent variation and hybridization cause the displacement of a sexually deceptive orchid species. AB - In the sexually deceptive orchid genus Ophrys, reproductive isolation is based on the specific attraction of males of a single pollinator species, mostly bees, by mimicking the female sex pheromone of this species. Changes in the floral odor can lead to hybridization, introgression, and possibly speciation. We investigated hybrid swarms of O. lupercalis and O. iricolor on Sardinia using behavioral, electrophysiological (GC-EAD), chemical, morphological, and genetic methods (AFLPs). In behavioral experiments, approximately 20% of the flowers from both species and hybrids were attractive to the "wrong" or both pollinator species. Analysis of the EAD-active hydrocarbons in the floral odor showed an overlap in the two species, whereby hybrid individuals could not be separated from O. iricolor. The genetic analysis confirmed the hybridization of the species. Plants of O. iricolor and hybrids are genetically indistinguishable and form an O. iricolor * lupercalis hybrid population. Remaining plants of O. lupercalis will possibly be displaced by the O. iricolor * lupercalis hybrid population in the future. Our study showed that in deceptive orchids, variation in the pollinator attracting cues, in this case, scent, can be the first step for speciation and at the same time cause the displacement of a species. PMID- 21632373 TI - Adaptive significance of self-fertilization in a hermaphroditic perennial, Trillium camschatcense (Melanthiaceae). AB - The evolution of self-fertilization from primarily outcrossing ancestors is one of the most common evolutionary transitions in plants; however, the ecological mechanisms that maintain self-fertilization have remained controversial. Theoretical studies suggest that selfing is advantageous over outcrossing in terms of genetic transmission and assurance of seed production under pollen limited circumstances. Trillium camschatcense is a herbaceous perennial distributed in Hokkaido and northern Honshu, Japan. Geographical variation in the breeding system (self-compatible, SC; or self-incompatible, SI) has been reported in populations in Hokkaido. Here, we used several SC and SI populations of T. camschatcense to investigate the adaptive significance and the evolutionary basis of self-fertilization. Pollination experiments and genetic analyses demonstrated that the potential availability of outcross pollen in SC populations was sufficient and that the number of pollen donors was equal to that of SI populations. However, despite the high availability of outcross pollen, the SC populations produced seeds predominantly by selfing and so underwent severe inbreeding depression. Although none of the suggested advantages for self fertilization were supported by our analyses, we propose two possible scenarios for the evolution of self-fertilization in T. camschatcense. PMID- 21632374 TI - Disentangling the causes of intrainflorescence variation in floral traits and fecundity in the hermaphrodite Silene acutifolia. AB - Inflorescence architecture directly determines variations in floral traits and fecundity. Disentangling these patterns of variation is crucial to understanding intraplant variation, which sometimes is directly attributed to competition for resources with developing fruits. The dichasial cymes of Silene acutifolia were experimentally manipulated in the field to analyze whether the declines in petal size, ovule number, fruit set, and seed/ovule ratio along the inflorescence are constrained by ontogenetic development or are phenotypically plastic in response to environmental changes. At the same time, the level of pollen deficit was measured on different positions of the dichasia. The results showed clearly that all measured variables were more influenced by architecture than by resource competition with developing fruits; the removal of central (basal) and primary lateral flowers in the dichasia did not increase either the measures of floral characters or fecundity. On the other hand, although most of the decline in fecundity was due to architectural effects, there was also a pollen limitation, dependent to some degree on inflorescence position, which was probably due to lower pollen availability in the population when secondary flowers are in the female phase. PMID- 21632375 TI - Relationships and evolution of matK in a group of leafless orchids (Corallorhiza and Corallorhizinae; Orchidaceae: Epidendroideae). AB - Corallorhizinae are a small group of Old and New World temperate orchids of which a core monophyletic group comprises Govenia, Cremastra, Aplectrum, Oreorchis and the leafless Corallorhiza, and which according to phylogenetic analysis of nuclear ITS and plastid matK sequences, are related in this way: (Govenia (Cremastra (Aplectrum (Oreorchis (Corallorhiza))))). This hypothesis is consistent with the progressive deletion of the trnK intron and matK ORF. Frameshift-resulting indels yield a predicted loss of translation for the critical "domain X" region of matK and are evidence that matK is a probable pseudogene in Aplectrum, Oreorchis, and Corallorhiza. Within Corallorhiza, a previous hypothesis based on plastid DNA restriction site analysis is confirmed, with the thickened-labellum C. striata group being sister to the thin-labellum remainder of the genus, within which the circumboreal C. trifida is sister to the remainder, which then comprise two further sister groups: C. maculata + C. bulbosa + C. mertensiana and C. odontorhiza + C. wisteriana. A close relationship between C. striata and the recently described Appalachian C. bentleyi is shown; in particular, C. bentleyi is more closely allied to a southern Mexican population of C. striata than it is to northern North American C. striata populations, suggesting that two lineages, each with Mexican and northern North American populations, exist within the C. striata group. PMID- 21632376 TI - Bayesian reconstruction of ancestral expression of the LEA gene families reveals propagule-derived desiccation tolerance in resurrection plants. AB - Desiccation tolerance is a complex trait that is broadly but infrequently present throughout the evolutionary tree of life. Desiccation tolerance has played a significant role in land plant evolution, in both the vegetative and reproductive life history stages. In the land plants, the late embryogenesis abundant (LEA) gene families are involved in both abiotic stress tolerance and the development of reproductive propagules. They are also a major component of vegetative desiccation tolerance. Phylogenies were estimated for four families of LEA genes from Arabidopsis, Physcomitrella, and the desiccation tolerant plants Tortula ruralis, Craterostigma plantagineum, and Xerophyta humilis. Microarray expression data from Arabidopsis and a subset of the Physcomitrella LEAs were used to estimate ancestral expression patterns in the LEA families and to evaluate alternative hypotheses for the origins of vegetative desiccation tolerance in the flowering plants. The results contradict the idea that vegetative desiccation tolerance in the resurrection angiosperms Craterostigma and Xerophyta arose through the co-option of genes exclusively related to stress tolerance, and support the propagule-derived origin of vegetative desiccation tolerance in the resurrection plants. PMID- 21632377 TI - The relationship between stem and branch wood specific gravity and the ability of each measure to predict leaf area. AB - A few trait axes that represent differential biomass allocation may summarize plant life-history strategies. Here we examine one of these axes described by wood specific gravity. Wood specific gravity represents the location of a species on a continuum of the rate of growth vs. the likelihood of mechanical failure, ranging from rapid volumetric growth/increased probability of mechanical failure to slow volumetric growth/decreased probability of mechanical failure. Wood specific gravity has been quantified primarily using three separate methods: a section from terminal branch, a section from the main stem or from a trunk wood core. What is unclear is how comparable these methods are and whether one or the other is a better predictor of other important plant traits such as leaf area. Here we measured stem and branch wood specific gravities from individual trees and shrubs in a tropical rain forest, quantified their relationship and determined their ability to predict leaf area. Stem and branch measures were highly correlated with each measure having a weak correlation with leaf area in trees and strong correlation with leaf area in shrubs. These results indicate that various methodologies for measuring wood specific gravity are comparable, and thus less destructive methods than are currently used are available to determine values for this important trait. PMID- 21632378 TI - The evolutionary relations of sunken, covered, and encrypted stomata to dry habitats in Proteaceae. AB - Sunken, covered, and encrypted stomata have been anecdotally linked with dry climates and reduced transpiration and therefore have been used to infer dry palaeoclimates from fossils. This study assesses the evolutionary and ecological associations of such stomatal protection in a model system-the diverse southern hemisphere family Proteaceae. Analyses were based on the morphology of over 1400 Australian, South African, New Caledonian, New Zealand, and South American species, anatomy of over 300 of these species, and bioclimatic data from all 1109 Australian species. Ancestral state reconstruction revealed that five or six evolutionary transitions explain over 98% of the dry climate species in the family, with a few other, minor invasions of dry climates. Deep encryption, i.e., stomata in deep pits, in grooves, enclosed by tightly revolute margins or strongly overarched by cuticle, evolved at least 11 times in very dry environments. Other forms of stomatal protection (sunken but not closely encrypted stomata, papillae, and layers of hairs covering the stomata) also evolved repeatedly, but had no systematic association with dry climates. These data are evidence for a strong distinction in function, with deep encryption being an adaptation to aridity, whereas broad pits and covered stomata have more complex relations to climate. PMID- 21632379 TI - A novel ascomycetous endophytic association in the rhizoids of the leafy liverwort family, Schistochilaceae (Jungermanniidae, Hepaticopsida). AB - Liverworts form diverse associations with endophytic fungi similar to mycorrhizas in vascular plants. Whereas the widespread occurrence of glomeromycotes in the basal liverwort lineages is well documented, knowledge of the distribution of ascomycetes and basidiomycetes in derived thalloid and leafy clades is more fragmented. Our discovery that the ramified and septate rhizoids of the Schistochilaceae, the sister group to all other ascomycete-containing liverworts, are packed with fungal hyphae prompted this study on the effects of the fungi on rhizoid morphology, host specificity, the cytology of the association, and a molecular analysis of the endophytes. Two species of Pachyschistochila and their fungi were grown axenically. Axenic rhizoids were unbranched and nonseptate. Reinfected with their own fungus and that from the other species, both Pachyschistochila species produced branched and septate rhizoids identical to those in nature. Woronin bodies and simple septa identified the fungus as an ascomycete referable, according to phylogenetic analyses of ITS sequences, to the Rhizoscyphus (Hymenoscyphus) ericae aggregate, also found in other liverwort ascomycete associations and in mycorrhizas in the Ericales. Healthy hyphae and host cytoplasm suggest that the Schistochila-fungus association reflects a balanced mutualistic relationship. The recent dating of the divergence of the Jungermanniales from the fungus-free Porellales in the Permian and the origins of the Schistochilaceae in the Triassic indicate that these associations in liverworts predate the appearance of the Ericales. PMID- 21632380 TI - Bryophyte diaspore bank: a genetic memory? Genetic structure and genetic diversity of surface populations and diaspore bank in the liverwort Mannia fragrans (Aytoniaceae). AB - Propagule banks are assumed to be able to store considerable genetic variability. Bryophyte populations are expected to rely more heavily on stored propagules than those of seed plants due to the vulnerability of the haploid gametophyte. This reliance has important implications for the genetic structure and evolutionary potential of surface populations. A liverwort, Mannia fragrans, was used to test whether the bryophyte diaspore bank functions as a "genetic memory." If a diaspore bank is capable of conserving genetic variability over generations, the levels of genetic diversity in the soil are expected to be similar or higher than at the surface. Surface and diaspore bank constituents of two populations of M. fragrans were investigated. Genetic structure and diversity measured as unbiased heterozygosity were analyzed using three ISSR markers. Similar genetic diversities were found in the soil (H(s) = 0.067) and at the surface (H(s)= 0.082). However, more haplotypes and specific haplotype lineages were present in soil samples. The results suggest that the bryophyte diaspore bank has an important role in accumulating genetic variability over generations and seasons. It is postulated that the role of the diaspore bank as a "genetic memory" is especially important in species of temporarily available habitats that have long lived spores and genetically variable populations. PMID- 21632381 TI - Evidence for "diminishing returns" from the scaling of stem diameter and specific leaf area. AB - Research indicates that increases in total leaf area (A(T)) may fail to keep pace with increases in total leaf mass (M(L)) across plants differing in size (e.g., as measured by stem diameter, D). This "diminishing returns" hypothesis predicts that the scaling exponent for A(T) vs. M(L) will be less than one and that the exponent for specific leaf mass (i.e., A(T) / M(L)) vs. D will be negative. These predictions were examined using data from 46 plants ranging between 0.125 cm <= D <= 0.485 m across 25 woody dicot species. Standardized major axis slopes were used to quantify scaling exponents and random effects models were used to quantify species and size effects on the numerical values of exponents. The exponents for A(T) vs. M(L) and A(T) / M(L) vs. D differed among species and different species groupings. In general, the exponent for A(T) vs. M(L) was less than one and the exponent for A(T) / M(L) vs. D was negative, as predicted. However, random effects models indicated that species effects overshadowed size effects, although size effects were statistically significant. The diminishing returns hypothesis therefore receives statistical support, i.e., although the numerical values of exponents are "species-dependent," they are less than unity, as predicted by theory. PMID- 21632383 TI - Local genetic structure in a North American epiphytic lichen, Ramalina menziesii (Ramalinaceae). AB - Epiphytic lichens possess unique life history traits that can have conflicting effects on genetic structure: symbiotic mutualism between a fungus with its algal or cyanobacterial photobiont, association with a host plant, and ability to reproduce sexually and asexually. Our study species, Ramalina menziesii, has small ascospores that can facilitate long-distance gene movement, and it is capable of clonal reproduction. The goals of this study are to test whether different haplotypes were differentially distributed across host plant species, to look for evidence of asexual vs. sexual reproduction, and to assess the local genetic structure of the population. We sampled individuals from multiple trees of three oak species in four lichen subpopulations within a savanna ecosystem. Using DNA sequence data from four fungal nuclear loci, we found no tendency for host specialization. Alleles were randomly distributed across subpopulations. The frequency of multilocus genotypes was consistent with a randomly mating population. Sexual reproduction involving relichenization appeared to be the predominant mode of reproduction of R. menziesii at this study site. We found no significant local genetic structure suggesting widespread gene flow at the local scale. The genetic structure of this lichen is comparable to that of widely distributed epiphytic plants. PMID- 21632382 TI - Mass flowering and parental death in the regeneration of Cerberiopsis candelabra (Apocynaceae), a long-lived monocarpic tree in New Caledonia1. AB - Monocarpy is rare among forest trees, and the selective conditions allowing this life history to evolve are poorly known. Here we examined the regeneration dynamics of a New Caledonian monocarpic tree species, Cerberiopsis candelabra, to better understand how postreproductive mortality and mass flowering might contribute to the success of this strategy. We investigated population size structures and seedling establishment patterns in 18 stands following the 2003 flowering event. We found little evidence of recent recruitment. Instead, population size structures suggested a history of one or more substantial recruitment events followed by recruitment failure, despite multiple flowering events in most populations. The canopy gap created by death of an individual parent appeared generally insufficient to allow seedling establishment, but mass flowering and consequent death of many adults appeared to enhance seedling recruitment by opening more of the canopy. Site means of seedling density below dead parents correlated strongly with light availability and with the proportional density of flowering trees. Therefore, we suggest that mass flowering and gregariousness may have been necessary preconditions for the evolution of monocarpy in this species. However, the degree to which regeneration relies on synchronous flowering and gregariousness vs. infrequent catastrophic disturbances remains uncertain. PMID- 21632384 TI - Genetic structure and diversity in Ramonda myconi (Gesneriaceae): effects of historical climate change on a preglacial relict species. AB - The importance of the Mediterranean Basin as a long-term reservoir of biological diversity has been widely recognized, although much less effort has been devoted to understanding processes that allow species to persist in this area. Ramonda myconi (Gesneriaceae) is a Tertiary relict plant species restricted to the NE Iberian Peninsula. We used RAPD and chloroplast markers to assess the patterns of genetic structure in eight mountain regions covering almost the full species range, to identify the main historical processes that have shaped its current distribution and to infer the number and location of putative glacial refugia. While no cpDNA variation was detected, the species had relatively high levels of RAPD variation. Maximum levels of diversity were found within populations (71%), but there was also a significant differentiation between geographical regions (20%) and among populations within regions (9%). A spatial AMOVA identified three main groups of populations, corresponding to previously recognized centers of endemism and species richness. In addition, we found a marked geographical pattern of decreasing genetic diversity and increasing population differentiation from west to east. Our results support a complex phylogeographic scenario in the Iberian Peninsula of "refugia-within-refugia" and suggest that the higher diversity observed in western regions might be associated with prolonged and more stable climatic conditions in this area during the Quaternary. PMID- 21632385 TI - Comparison of population genetic diversity between a rare, narrowly distributed species and a common, widespread species of Alnus (Betulaceae). AB - Comparisons of population genetic diversity between related rare and widespread species provide valuable insights to the consequences of rarity and are critical for conservation planning. Population genetic diversity of A. maritima, a rare species, was compared with its common, widespread congener A. serrulata to evaluate the impacts of small population size and high isolation on genetic diversity in A. maritima and to provide population genetic data to be used in conservation planning for A. maritima. Genetic data were also used to evaluate whether the disjunct distribution of A. maritima was due to range reduction or anthropogenic dispersal. Genetic diversity was lower in A. maritima (H(e) = 0.217) than in A. serrulata (H(e) = 0.268), and there is also higher inbreeding within A. maritima populations (f = 0.483) than A. serrulata populations (f = 0.269). The partitioning of genetic variation was also higher among A. maritima populations (Theta = 0.278), but not significantly different from that of A. serrulata (Theta = 0.197). Significant genetic differences among A. maritima populations support using local populations as seed sources for regional conservation efforts. The results also indicate that the highly disjunct distribution of A. maritima is due to natural range reduction in the past and not anthropogenic establishment of Oklahoma and Georgia populations. PMID- 21632386 TI - Hybridization between the escaped Rosa rugosa (Rosaceae) and native R. blanda in eastern North America. AB - Rosa rugosa, a vigorous ornamental shrub introduced from Asia in the 19th century, is now naturalized in coastal northeastern North America, where it occasionally grows in sympatry with the native R. blanda. To document hybridization between these species, evaluate its extent across the area of sympatry, and examine the use of morphology as a field monitoring tool, we sampled 179 individuals of parental species and putative hybrids in 13 pure and 11 mixed populations. We developed allele-specific primers to assay single nucleotide polymorphisms (SNPs) markers from one chloroplast region and four low copy nuclear introns. Our results revealed frequent bidirectional hybridization and infrequent introgression in sympatric populations of these species. The recurrent presence of F(1) hybrids in mixed populations indicated the weakness of early-acting reproductive barriers. Morphological data were concordant with molecular data and provided additional evidence for the presence of a few backcrosses. Morphological analyses yielded diagnostic characters for identifying hybrids and monitoring the hybrid zone. Such hybridization could ultimately lead to the genetic assimilation of R. blanda in mixed populations and to the formation of invasive hybrid genotypes, a phenomenon that is of economic and ecological concern because of the increasing number of exotic species worldwide. PMID- 21632387 TI - From annuals to perennials: phylogeny of subtribe Castillejinae (Orobanchaceae). AB - Variation in life history strategies is a fundamental question in evolutionary biology, and the cooccurrence of annual and perennial habits in Castilleja and Castillejinae provides the opportunity to study the evolution of plant life history in a phylogenetic context. Molecular phylogenetic analysis of two chloroplast (rps16 and trnL/F) and two nuclear ribosomal (internal and external transcribed spacers) DNA regions support the monophyly of subtribe Castillejinae (Orobanchaceae). A well-supported phylogeny of the six genera (Castilleja [~180 spp.], Clevelandia [1 sp.], Cordylanthus [18 spp.], Ophiocephalus [1 sp.], Orthocarpus [9 spp.], and Triphysaria [5 spp.]) comprising the subtribe is presented, and morphological synapomorphies are identified for the major lineages recovered. Orthocarpus and Triphysaria are both monophyletic; Cordylanthus is biphyletic. Clevelandia and Ophiocephalus are derived from within Castilleja. The perennial Castilleja clade (~160 spp.) is derived from a grade of annual taxa including Castilleja sect. Oncorhynchus (16 spp.), Cordylanthus, Orthocarpus, and Triphysaria. This suggests that the perennial habit evolved a single time from an annual ancestral lineage that persisted throughout the diversification of Castillejinae, contrary to classical interpretations of life history evolution in plants. Given the prevalence of polyploidy among perennial Castilleja species, perenniality may have played an important role in the origin and establishment of polyploidy in Castilleja. PMID- 21632388 TI - Phylogeny of marattioid ferns (Marattiaceae): inferring a root in the absence of a closely related outgroup. AB - Closely related outgroups are optimal for rooting phylogenetic trees; however, such ideal outgroups are not always available. A phylogeny of the marattioid ferns (Marattiaceae), an ancient lineage with no close relatives, was reconstructed using nucleotide sequences of multiple chloroplast regions (rps4 + rps4-trnS spacer, trnS-trnG spacer + trnG intron, rbcL, atpB), from 88 collections, selected to cover the broadest possible range of morphologies and geographic distributions within the extant taxa. Because marattioid ferns are phylogenetically isolated from other lineages, and internal branches are relatively short, rooting was problematic. Root placement was strongly affected by long-branch attraction under maximum parsimony and by model choice under maximum likelihood. A multifaceted approach to rooting was employed to isolate the sources of bias and produce a consensus root position. In a statistical comparison of all possible root positions with three different outgroups, most root positions were not significantly less optimal than the maximum likelihood root position, including the consensus root position. This phylogeny has several important taxonomic implications for marattioid ferns: Marattia in the broad sense is paraphyletic; the Hawaiian endemic Marattia douglasii is most closely related to tropical American taxa; and Angiopteris is monophyletic only if Archangiopteris and Macroglossum are included. PMID- 21632389 TI - Patterns of cell division in the filamentous Desmidiaceae, close green algal relatives of land plants. AB - Patterns of cell division and cross wall formation vary among the charophytes, green algae closely related to land plants. One group of charophytes, the conjugating green algae (Zygnematophyceae), is species-rich and is known to vary substantially in the mode of cell division, but the details of these cell division patterns and their phylogenetic distribution remain poorly understood. We studied cross wall development in filamentous Desmidiaceae (a clade of conjugating green algae) using differential interference contrast and fluorescence light microscopy. All strains investigated had centripetal encroachment of a septum, but with several different developmental patterns. In most cases, cell wall formation was delayed with respect to the Cosmarium-type of cell division, and the cross wall was modified considerably after deposition in a manner specific to the particular clade of filamentous desmids. These characteristics were mapped on a phylogeny estimated from a data set of two organellar genes, and the evolutionary implications of the character state distribution were evaluated. The data suggest a complex history of evolution of cell division in this lineage and also imply that Desmidium and Spondylosium are polyphyletic. These results indicate that many features of the cell shape are determined at the time of cell division in conjugating green algae. PMID- 21632390 TI - Immunocytochemical characterization of tension wood: Gelatinous fibers contain more than just cellulose. AB - Gelatinous fibers (G-fibers) are the active component of tension wood. G-fibers are unlike traditional fiber cells in that they possess a thick, nonlignified gelatinous layer (G-layer) internal to the normal secondary cell wall layers. For the past several decades, the G-layer has generally been presumed to be composed nearly entirely of crystalline cellulose, although several reports have appeared that disagreed with this hypothesis. In this report, immunocytochemical techniques were used to investigate the polysaccharide composition of G-fibers in sweetgum (Liquidambar styraciflua; Hamamelidaceae) and hackberry (Celtis occidentalis; Ulmaceae) tension wood. Surprisingly, a number of antibodies that recognize arabinogalactan proteins and RG I-type pectin molecules bound to the G layer. Because AGPs and pectic mucilages are found in other plant tissues where swelling reactions occur, we propose that these polymers may be the source of the contractile forces that act on the cellulose microfibrils to provide the tension force necessary to bend the tree trunk. PMID- 21632391 TI - {gamma}-Tubulin and microtubule organization during meiosis in the liverwort Ricciocarpus natans (Ricciaceae). AB - Extant liverworts are "living fossils" considered sister to all other plants and as such provide clues to the evolution of the microtubule organizing center (MTOC) in anastral cells. This report is the first on microtubule arrays and their gamma-tubulin-nucleating sites during meiosis in a member of the Ricciales, a specialized, species-rich group of complex thalloid (marchantioid) liverworts. In meiotic prophase, gamma-tubulin becomes concentrated at several sites adjacent to the nuclear envelope. Microtubules organized at these foci give rise to a multipolar prometaphase spindle. By metaphase I, the spindle has matured into a bipolar structure with truncated poles. In both first and second meiosis, gamma tubulin forms box-like caps at the spindle poles. gamma-Tubulin moves from spindle poles to the proximal surfaces of telophase chromosomes where interzonal microtubules are nucleated. Although a phragmoplast is organized, no cell plate is deposited, and second division occurs simultaneously in the undivided sporocyte. gamma-Tubulin surrounds each of the tetrad nuclei, and phragmoplasts initiated between both sister and nonsister nuclei direct simultaneous cytokinesis. The overall pattern of meiosis (unlobed polyplastidic sporocytes, nuclear envelope MTOC, multipolar spindle origin, spindles with box-like poles, and simultaneous cytokinesis) more closely resembles that of Conocephalum than other marchantiod liverworts. PMID- 21632392 TI - Responses of two bunchgrasses to nitrogen addition in tallgrass prairie: the role of bud bank demography. AB - Growth of tallgrass prairie plants, many of which maintain substantial bud banks, can be limited by nitrogen (N), water, and/or light. We hypothesized that tallgrass prairie plants respond to increases in N through demographic effects on the bud bank. We tested the effects of a pulse of N on (1) bud bank demography, (2) plant reproductive allocation, and (3) ramet size. We parameterized matrix models, considering each genet as a population of plant parts. Nitrogen addition significantly impacted bud bank demography in two subdominant species of bunchgrass: Sporobolus heterolepis (a C(4) grass) and Koeleria macrantha (a C(3) grass), but had no effect on the size of individual ramets. Emergence from the bud bank and ramet population growth rates (lambda) were significantly higher in S. heterolepis genets that received supplemental N. Nitrogen addition also affected the bud demography of K. macrantha, but N addition decreased rather than increased lambda. Prospective and retrospective demographic analyses indicated that bud bank dynamics were the most important demographic processes driving plant responses to nutrient availability. Thus, the variation in productivity in these tallgrass prairie species is driven principally by the demography of the bud bank rather than by the physiology and growth of aboveground tillers. Improved understanding of bud bank dynamics may lead to improved predictive models of grassland responses to environmental changes such as altered N deposition and precipitation. PMID- 21632393 TI - Variation in resource acquisition and utilization traits between native and invasive perennial forbs. AB - Understanding the functional traits that allow invasives to outperform natives is a necessary first step in improving our ability to predict and manage the spread of invaders. In nutrient-limited systems, plant competitive ability is expected to be closely tied to the ability of a plant to exploit nutrient-rich microsites and use these captured nutrients efficiently. The broad objective of this work was to compare the ability of native and invasive perennial forbs to acquire and use nutrients from nutrient-rich microsites. We evaluated morphological and physiological responses among four native and four invasive species exposed to heterogeneous (patch) or homogeneous (control) nutrient distribution. Invasives, on average, allocated more biomass to roots and allocated proportionately more root length to nutrient-rich microsites than did natives. Invasives also had higher leaf N, photosynthetic rates, and photosynthetic nitrogen use efficiency than natives, regardless of treatment. While these results suggest multiple traits may contribute to the success of invasive forbs in low-nutrient environments, we also observed large variation in these traits among native forbs. These observations support the idea that functional trait variation in the plant community may be a better predictor of invasion resistance than the functional group composition of the plant community. PMID- 21632394 TI - Fossil cyclanthus (cyclanthaceae, pandanales) from the eocene of Germany and England. AB - The first known fossil fruits and seeds of Cyclanthaceae are described here. Cyclanthus messelensis sp. nov., from the Middle Eocene of Messel, Germany, has discoidal fruiting cycles up to 6 cm in diameter, with a central hole, radiating fiber strands, a thickened outer rim, and paratetracytic stomata. In situ seeds are up to 2 mm long, with an elongate micropylar end, a chalazal neck, and adpressed bands. The Messel fruits and seeds are nearly identical to those of Cyclanthus, differing in minor details of cuticular structure and seeds. The exceptional preservation at Messel (including in situ and isolated seeds) has also allowed us to establish the taxonomic affinity of isolated seeds ('Scirpus' lakensis) that are spatially and temporally widespread in the late Early and early Middle Eocene of southern England. Cyclanthus lakensis comb. nov. is described here as a morphotaxon for isolated fossil Cyclanthus seeds, preserved only as cuticular envelopes. Cyclanthus is another example of links between Eocene Europe and Recent South American floras because it is found today only from Mexico to South America. This material represents the first fossil fruits and seeds of Cyclanthus, which clearly was once growing in the Paleogene of the Old World. PMID- 21632395 TI - Detecting the influence of ornamental Berberis thunbergii var. atropurpurea in invasive populations of Berberis thunbergii (Berberidaceae) using AFLP1. AB - Japanese barberry (Berberis thunbergii DC.) is a widespread invasive plant that remains an important landscape shrub represented by ornamental, purple-leaved forms of the botanical variety atropurpurea. These forms differ greatly in appearance from feral plants, bringing into question whether they contribute to invasive populations or whether the invasions represent self-sustaining populations derived from the initial introduction of the species in the late 19th century. In this study we used amplified fragment length polymorphism (AFLP) markers to determine whether genetic contributions from B. t. var. atropurpurea are found within naturalized Japanese barberry populations in southern New England. Bayesian clustering of AFLP genotypes and principal coordinate analysis distinguished B. t. var. atropurpurea genotypes from 85 plants representing five invasive populations. While a single feral plant resembled B. t. var. atropurpurea phenotypically and fell within the same genetic cluster, all other naturalized plants sampled were genetically distinct from the purple-leaved genotypes. Seven plants from two different sites possessed morphology consistent with Berberis vulgaris (common barberry) or B. *ottawensis (B. thunbergii * B. vulgaris). Genetic analysis placed these plants in two clusters separate from B. thunbergii. Although the Bayesian analysis indicated some introgression of B. t. var. atropurpurea and B. vulgaris, these genotypes have had limited influence on extant feral populations of B. thunbergii. PMID- 21632396 TI - How does secondary pollen presentation affect the fitness of Polygala vayredae (Polygalaceae)? AB - Secondary pollen presentation is the relocation and presentation of pollen in floral structures (termed pollen presenters) other than the anthers. These pollen presenters are often found close to the stigma and have been hypothesized to increase the accuracy of pollen transfer, although no experimental studies have been done. We examined the function of the pollen presenter and its efficiency in pollen dispersal, female fitness, and the degree of interference created by self pollen in the shrublet Polygala vayredae, an insect-pollinated species with secondary pollen presentation. Herkogamy, a mechanism generally involved in the reduction of self-interference, was also evaluated. Significant pollen was lost (49% of total pollen) during the secondary relocation in the pollen presenter. However, pollen was exported from the pollen presenter, and subsequent pollen losses were similar to those in species with primary pollen presentation. Despite the presence of a self-incompatibility system, the numbers of developed pollen tubes as well as fruit and seed production were significantly reduced by the self pollen interference created at the stigmatic papillae level. The extent of herkogamy correlated positively with female fitness. The secondary pollen presentation mechanism may in fact be an accurate system for pollen transport, but it may also have its costs. Further comparative studies involving species with primary and secondary pollen presentation are needed to fully understand the advantages and disadvantages of secondary pollen presentation. PMID- 21632397 TI - Hybridization between invasive Spartina densiflora (Poaceae) and native S. foliosa in San Francisco Bay, California, USA. AB - Rapid evolution in contemporary time can result when related species, brought together through human-aided introduction, hybridize. The significant evolutionary consequences of post-introduction hybridization range from allopolyploid speciation to extinction of species through genetic amalgamation. Both processes are known to occur in the perennial cordgrass genus, Spartina. Here we report the existence of a third recent Spartina hybridization, discovered in 2002, between introduced S. densiflora and native S. foliosa in San Francisco Bay, California, USA. We used nuclear and chloroplast DNA analysis and nuclear DNA content with chromosome counts to examine plants of morphology intermediate between S. densiflora and S. foliosa in a restored marsh in Marin County, California. We found 32 F(1) diploid hybrids and two triploid plants, all having S. densiflora and S. foliosa as parents; there is also evidence of a genetic contribution of S. alterniflora in some hybrids. None of these hybrids set germinable seed. In 2007 we found a hybrid over 30 miles away in a marsh where both parental species occurred, suggesting hybridization may not be a localized phenomenon. The presence of diploid and triploid hybrids is important because they indicate that several avenues existed that may have given rise to a new allopolyploid species. However, such an event is now unlikely because all hybrids are targets of eradication efforts. PMID- 21632398 TI - Origin and evolution of the northern hemisphere disjunction in the moss genus Homalothecium (Brachytheciaceae). AB - Competing hypotheses that rely either on a stepping-stone dispersal via the North Atlantic or the Bering land bridges, or more recent transoceanic dispersal, have been proposed to explain the disjunct distribution of Mediterranean flora in southern Europe and western North America. These hypotheses were tested with molecular dating using a phylogeny of the moss genus Homalothecium based on ITS, atpB-rbcL, and rpl16 sequence data. The monophyly of two main lineages in Western Palearctic (Europe, central Asia and north Africa) and North America is consistent with the ancient vicariance hypothesis. The monophyly of Madeiran H. sericeum accessions supports the recognition of the Macaronesian endemic H. mandonii. A range of absolute rates of molecular evolution documented in land plants was used as probabilistic calibration prior by a Bayesian inference implementing a relaxed-clock model to derive ages for the nodes of interest. Our age estimates for the divergence of the American and Western Palearctic Homalothecium clade (5.7 Ma, IC 3.52-8.26) and the origin of H. mandonii (2.52 Myr IC 0.86-8.25) are not compatible with the ancient vicariance hypothesis. Age estimates suggests that species distributions result from rare instances of dispersal and subsequent sympatric diversification. The calibrated phylogeny indicates that Homalothecium has undergone a fast radiation during the last 4 Myr, which is consistent with the low levels of morphological divergence among sibling species. PMID- 21632399 TI - A phylogenetic delimitation of the "Sphagnum subsecundum complex" (Sphagnaceae, Bryophyta). AB - A seemingly obvious but sometimes overlooked premise of any evolutionary analysis is delineating the group of taxa under study. This is especially problematic in some bryophyte groups because of morphological simplicity and convergence. This research applies information from nucleotide sequences for eight plastid and nuclear loci to delineate a group of northern hemisphere peat moss species, the so-called Sphagnum subsecundum complex, which includes species known to be gametophytically haploid or diploid (i.e., sporophytically diploid-tetraploid). Despite the fact that S. subsecundum and several species in the complex have been attributed disjunct ranges that include all major continents, phylogenetic analyses suggest that the group is actually restricted to Europe and eastern North America. Plants from western North America, from California to Alaska, which are morphologically similar to species of the S. subsecundum complex in eastern N. America and Europe, actually belong to a different deep clade within Sphagnum section Subsecunda. One species often considered part of the S. subsecundum complex, S. contortum, likely has a reticulate history involving species in the two deepest clades within section Subsecunda. Nucleotide sequences have a strong geographic structure across the section Subsecunda, but shallow tip clades suggest repeated long-distance dispersal in the section as well. PMID- 21632400 TI - Strait of Gibraltar: an effective gene-flow barrier for wind-pollinated Carex helodes (Cyperaceae) as revealed by DNA sequences, AFLP, and cytogenetic variation. AB - The Strait of Gibraltar is the most important barrier disconnecting the landmasses of Europe and Africa on the western Mediterranean extreme. Carex helodes is a wind-pollinated species endemic to the western Mediterranean. Because molecular and cytogenetic data allow the inference of its evolutionary history, we analyzed variations in chromosome number, including meiotic chromosome behavior, amplified fragment length polymorphism (AFLP) fingerprints, and nucleotide substitutions in plastid and nuclear DNA sequences. Cytogeographic results showed that the African populations have stabilized at a single chromosome number of 2n = 74, whereas the most frequent cytotype in Iberia is 2n = 72. Phylogenetic reconstructions of 17 sequences from nine closely related species revealed that C. helodes is monophyletic and that the Moroccan populations are embedded in the Iberian lineages. The haplotype network is also consistent with a European origin of the northern African haplotype. AFLP analysis also revealed hierarchical levels of genetic variation compatible with a founder effect process responsible for the African populations. All sources of evidence support the hypothesis that the Strait of Gibraltar has been an effective gene-flow barrier, generating two isolated evolutionary lineages after their dispersal. Recent connections between the two lineages appear unlikely, whereas active gene flow occurs among populations within the two lineages. PMID- 21632401 TI - Phylogenetic, morphological, and chemotaxonomic incongruence in the North American endemic genus Echinacea. AB - The study of recently formed species is important because it can help us to better understand organismal divergence and the speciation process. However, these species often present difficult challenges in the field of molecular phylogenetics because the processes that drive molecular divergence can lag behind phenotypic divergence. In the current study we show that species of the recently diverged North American endemic genus of purple coneflower, Echinacea, have low levels of molecular divergence. Data from three nuclear loci and two plastid loci provide neither resolved topologies nor congruent hypotheses about species-level relationships. This lack of phylogenetic resolution is likely due to the combined effects of incomplete lineage sorting, hybridization, and backcrossing following secondary contact. The poor resolution provided by molecular markers contrasts previous studies that found well-resolved and taxonomically supported relationships from metabolic and morphological data. These results suggest that phenotypic canalization, resulting in identifiable morphological species, has occurred rapidly within Echinacea. Conversely, molecular signals have been distorted by gene flow and incomplete lineage sorting. Here we explore the impact of natural history on the genetic organization and phylogenetic relationships of Echinacea. PMID- 21632402 TI - Ontogeny in terminal buds of Abies nordmanniana (Pinaceae) characterized by ubiquitin. AB - Meristematic activity in the bud meristem of Abies nordmanniana was visualized by ubiquitin immunohistochemical localization from before bud break and throughout shoot expansion. Ubiquitin was detected in meristematic cells either in the cytosol or nucleus, or both, depending on tissue type and developmental stage. During winter dormancy, ubiquitin was only observed in the protodermal/hypodermal layers, but at bud break in mid May, the signal expanded to the entire shoot tip. At the end of May, a clear zonation in ubiquitin localization appeared that lasted about one month. Throughout this period, ubiquitin was barely detectable in a central group of cells that might indicate an organizing center with stem cells. At the end of June, coinciding with the transition from scale leaf to needle primordia production, ubiquitin again was more prevalent in the peripheral cell layers. During shoot expansion, a strong ubiquitin signal developed in the axil of all needles. Most of these signals later disappeared, except for those few axils where buds actually developed. A strong ubiquitin signal was also observed in cells lining the young resin ducts. Our data showed that ubiquitin may be used as a marker for metabolic activity associated with seasonal development in the apical meristem. PMID- 21632403 TI - Development of the axillary bud complex in Echinocystis lobata (Cucurbitaceae): interpreting the cucurbitaceous tendril. AB - In the Cucurbitaceae, the tendrils, coiling organs used for climbing and mechanical support, are part of an axillary bud complex (ABC). Although the morphological nature of tendrils and the branching pattern of the ABC in the Cucurbitaceae have been much studied, their homology remains unresolved, with hypothesized candidates being the leaf, flower, stem, or stem-leaf combination. We used Echinocystis lobata as a model to study the early ontogeny of the ABC with epi-illumination microscopy and serial resin sections. The ABC produces four structures (proximal to distal, relative to the subtending leaf) as the result of two successive subdivisions: an inflorescence of staminate flowers, a solitary pistillate flower, a lateral bud, and a tendril. The first separates the tendril primordium from the continuation of the ABC, and the second separates the staminate inflorescence and the ABC. The pistillate flower apparently forms between the staminate inflorescence and the lateral bud. Because there is no subtending leaf during these subdivisions and the first lateral appendages in the resulting primordia arise in the same plane, we conclude that the tendril and other organs formed by the ABC are lateral branches of equal morphological value. This study is the basis for continuing comparative and functional morphological studies. PMID- 21632404 TI - Changes in cell wall ultrastructure induced by sudden flooding at 25{degrees}C in Pisum sativum (Fabaceae) primary roots. AB - Cellular degeneration is essential for many developmental and stress acclimation processes. Undifferentiated parenchymatous cells in the central vascular cylinder of pea primary roots degenerate under hypoxic conditions created by flooding at temperatures >15 degrees C, forming a long vascular cavity that seems to provide a conduit for longitudinal oxygen transport in the roots. We show that specific changes in the cell wall ultrastructure accompanied previously detected cytoplasmic and organellar degradation in the cavity-forming roots. The degenerating cells had thinner primary cell walls, less electron-dense middle lamellae, and less abundant cell wall homogalacturonans in altered patterns, compared to healthy cells of roots grown under cold, nonflooded conditions. Cellular breakdown and changes in wall ultrastructure, however, remained confined to cells within a 50-MUm radius around the root center, even after full development of the cavity. Cells farther away maintained cellular integrity and had signs of wall synthesis, perhaps from tight regulation of wall metabolism over short distances. These observations suggest that the cell degeneration might involve programmed cell death. We also show that warm, nonflooded or cold, flooded conditions that typically do not induce vascular cavity formation can also induce variations in cell wall ultrastructure. PMID- 21632405 TI - Major cytogenetic landmarks and karyotype analysis inDaucus carota and other Apiaceae. AB - Karyotype analysis provides insights into genome organization at the chromosome level and into chromosome evolution. Chromosomes were marked for comparative karyotype analysis using FISH localization of rDNA genes for the first time in Apioideae species including taxa of economic importance and several wild Daucus relatives. Interestingly, Daucus species did not vary in number of rDNA loci despite variation in chromosome number (2n = 18, 20, 22, and 44) and previous publications suggesting multiple loci. All had single loci for both 5S and 18S 25S (nucleolar organizing region) rDNA, located on two different chromosome pairs. The 5S rDNA was on the short arm of a metacentric chromosome pair in D. crinitus (2n = 22) and D. glochidiatus (2n = 44) and on the long arm of a metacentric pair in other Daucus species, suggesting possible rearrangement of this chromosome. For other Apiaceae, from two (Apium graveolens), to three (Orlaya grandiflora), to four (Cuminum cyminum) chromosomes had 18S-25S rDNA sites. Variability for number and position of the 5S rDNA was also observed. FISH signals enabled us to identify 20-40% of the chromosome complement among species examined. Comparative karyotype analysis provides insights into the fundamental aspects of chromosome evolution in Daucus. PMID- 21632406 TI - Patterns of introduction and diversification of Vanilla planifolia (Orchidaceae) in Reunion Island (Indian Ocean). AB - The cultivated species Vanilla planifolia is a typical example of a crop introduced from its area of origin (America) to new regions where natural pollinators are absent. Although the Vanilla cultivars are exclusively vegetatively propagated, a high degree of phenotypic variation is observed among the cultivars in their introduction areas such as Reunion Island. To test several hypotheses explaining this variation-different introduction events, somatic mutations and sexual reproduction (through manual pollination)-we used AFLP markers to elucidate the patterns of introduction of V. planifolia. Most of the accessions cultivated in the world were derived from a single accession, possibly the Mexican cultivar Mansa. The patterns of diversification of this cultivated species were also studied and compared with other cultivated (V. tahitensis) and wild (V. pompona and V. bahiana) species. Except for one particular phenotype ('Aiguille'), which may come from sexual reproduction, cultivated accessions exhibit very low levels of genetic diversity. They have evolved by the accumulation of point mutations through vegetative multiplication. The genetic diversity revealed could not explain the phenotypic diversity, which may be related to epigenetics or polyploidy. This new understanding of the basis of genetic diversity of vanilla may assist to improve management of genetic resources. PMID- 21632407 TI - Remarkable nutritional diversity of basidiomycetes in the Corticiales, including a new foliicolous species of Marchandiomyces (anamorphic Basidiomycota, Corticiaceae) from Australia. AB - Fungi in the basidiomycete order Corticiales are remarkably diverse nutritionally, including a variety of saprotrophs, plant and fungal pathogens, and lichen-forming fungi. Tracing the origin of this diversity depends on a clearer understanding of the phylogenetic relationships of fungi in the order. One of its core members is the genus Marchandiomyces, originally established for lichen pathogens that form orange or coral bulbils. We describe here a new species in the genus, M. marsonii sp. nov., which is unusual in its appearance, habit, and geographic provenance. It is foliicolous on leaves of Pandanus (screw pines, Pandanaceae) and produces flattened, coral bulbils resembling apothecia of the ascomycete genus Orbilia. It is also the first member of the genus to be collected from Australia. An isolate of the new fungus and several additional cultures of related plant pathogenic fungi were obtained and investigated phylogenetically using parsimony, likelihood, and Bayesian analyses of nuclear small and large subunit ribosomal sequences. Our phylogeny makes clear that Marchandiomyces species and their close relatives contribute significantly to the ecological diversity of the Corticiales and that this diversity is derived mainly from lignicolous ancestors. PMID- 21632408 TI - Fruits of icacinaceae (tribe iodeae) from the late paleocene of Western north america. AB - The Icacinaceae occur pantropically today, but are well represented by fossil fruits of the warm Early Middle Eocene, when tropical plants that currently occupy low latitudes were more widely distributed in higher latitudes. Members of this family are first known in the Late Cretaceous; however, fossil fruits of tribe Iodeae are quite rare before the Eocene. In this paper we describe the first formally recognized Late Paleocene icacinaceous taxa from western North America. We name two new species of Icacinicarya based on anatomically preserved fruits and establish a new genus, Icacinicaryites, for impressions with a strong similarity to Icacinicarya that lack anatomical preservation. These new records from the Almont/Beicegel Creek flora in North Dakota and several localities in Wyoming, Colorado, and Montana complement records known from the Early Eocene of England and document an increased diversity of Iodeae and related forms in the Paleogene of western North America. PMID- 21632409 TI - To grow or to seed: ecotypic variation in reproductive allocation and cone production by young female Aleppo pine (Pinus halepensis, Pinaceae). AB - Age and size at the first reproduction and the reproductive allocation of plants are linked to different life history strategies. Aleppo pine only reproduces through seed, and, as such, early female reproduction confers high fitness in its infertile highly fire-prone habitats along the Mediterranean coast because life expectancy is short. We investigated the extent of ecotypic differentiation in female reproductive allocation and examined the relation between early female reproduction and vegetative growth. In a common-garden experiment, the threshold age and size at first female reproduction and female reproductive allocation at age seven differed significantly among Aleppo pine provenances of ecologically distinct origin. Significant correlations among reproductive features of the provenances and the ecological traits of origin were found using different analytical tools. In nonlinear models of cone counts vs. stem volume, medium sized trees (not the largest trees) produced the highest cone yield, confirming that, at the individual level, early female reproduction is incompatible with fast vegetative growth. The contribution of founder effects and adaptation to contrasting fire regimes may be confounding factors. But considering all traits analyzed, the geographical patterns of resource allocation by Aleppo pine suggest ecotypic specialization for either resource-poor (favoring early reproduction) or resource-rich (favoring vegetative growth) habitats. PMID- 21632410 TI - A smaller Macadamia from a more vagile tribe: inference of phylogenetic relationships, divergence times, and diaspore evolution in Macadamia and relatives (tribe Macadamieae; Proteaceae). AB - Tribe Macadamieae (91 spp., 16 genera; Proteaceae) is widespread across the southern hemisphere on all major fragments of Gondwana except New Zealand and India. Macadamia is cultivated outside its natural range as a "nut" crop (notably in Hawaii, where it is the principal orchard crop). We sampled seven DNA regions and 53 morphological characters from the tribe to infer its phylogeny and address the common assumption that the distribution of the extant diversity of the tribe arose by the rafting of ancestors on Gondwanan fragments. Macadamia proves to be paraphyletic with respect to the African genus Brabejum, the South American genus Panopsis, and the Australian species Orites megacarpus. We erect two new generic names, Nothorites and Lasjia, to produce monophyly at that rank. The earliest disjunctions in the tribe are inferred to be the result of long-distance dispersal out of Australia (with one possible exception), rather than vicariance. Evolution of tardy fruit dehiscence is correlated with these dispersals, and the onset of the Antarctic Circumpolar Current (ACC) precedes them. We suggest that the ancestors of extant diversity arrived on their respective continents via the ACC, and we recognize that this is a mechanism precluded, rather than facilitated, by Gondwana's terrestrial continuity. PMID- 21632411 TI - Phylogenetic analyses of two mitochondrial metabolic genes sampled in parallel from angiosperms find fundamental interlocus incongruence. AB - Plant molecular phylogeneticists have supported an analytical approach of combining loci from different genomes, but the combination of mitochondrial sequences with chloroplast and nuclear sequences is potentially problematic. Low substitution rates in mitochondrial genes should decrease saturation, which is especially useful for the study of deep divergences. However, individual mitochondrial loci are insufficiently informative, so that combining congruent loci is necessary. For this study atp1 and cox1 were selected, which are of similar lengths, encode components of the respiratory pathway, and generally lack introns. Thus, these genes might be expected to have similar functional constraints, selection pressures, and evolutionary histories. Strictly parallel sampling of 52 species was achieved as well as six additional composite terminals with representatives from the major angiosperm clades. However, analyses of the separate loci produced strongly incongruent topologies. The source of the incongruence was investigated by validating sequences with questionable affinities, excluding RNA-edited nucleotides, deleting taxa with unexpected phylogenetic associations, and comparing different phylogenetic methods. However, even after potential artifacts were addressed and sites and taxa putatively associated with conflict were excluded, the resulting gene trees for the two mitochondrial loci were still substantially incongruent by all measures examined. Therefore, combining these loci in phylogenetic analysis may be counterproductive to the goal of fully resolving the angiosperm phylogeny. PMID- 21632412 TI - Multiple hybridization in the Aristolochia kaempferi group (Aristolochiaceae): evidence from reproductive isolation and molecular phylogeny. AB - Hybridization via distributional changes should be an important factor for plant speciation. Previous cpDNA analyses of the Aristolochia kaempferi group, comprising six taxa in East Asia, showed a distinct phylogeographic structure resulting from distributional changes brought about by paleoclimatic oscillations. However, the cpDNA phylogeny was incongruent with morphologically defined taxa. To explore the evolutionary processes responsible for the inconsistency between cpDNA and morphology, we made artificial crosses and performed phylogenetic analyses using multiple nuclear markers. All crosses among different taxa or cpDNA clades set fruit, if crossing direction is not considered. The five nuclear phylogenies mostly did not support either the taxa or the cpDNA clades. A combined analysis of cpDNA and the PI exon revealed the two major lineages in the group, lacking a prezygotic isolating barrier between them. However, an asymmetric prezygotic isolating barrier occurs between populations of the Japanese main islands and of other areas that belong to different cpDNA subclades. It seems reasonable to conclude that the development of a prezygotic isolating mechanism is not necessarily proportional to the degree of genetic divergence. These results suggested that species boundaries within the group are blurred due to speciational processes associated with multiple hybridization and introgression resulting from repeated contacts among differentiated populations. PMID- 21632413 TI - Phyllotactic pattern is altered in the transition to flowering in the early ears of Zea mays landrace chapalote (Poaceae). AB - The origin of polystichy in the maize ear and central tassel spike continues to challenge our understanding of evolution in this important crop species. In this paper we tested the hypothesis that the change in phyllotaxy occurs in the region of husk leaf production before the transition to reproductive growth. Young ear or presumptive ear primordia were dissected to examine the transition from distichous husk leaves below the ear through spiral phyllotaxy to the polystichous arrangement of spikelet pair primordia in the young ear. Serial transverse sections were used to document the thickness of successive disks of insertion of lateral primordia and to reconstruct the path of procambial differentiation. The transition in phyllotaxy, though variable, typically occurs in the vegetative zone and is associated with periodic heterogeneity in the thickness of leaf bases and a delay in the development of waves of procambial differentiation into the base of the young ear. PMID- 21632414 TI - Biogeographic and phylogenetic patterns in diversity of liverwort-associated endophytes. AB - Liverworts harbor diverse fungi, including endophytes, in their healthy tissues. To address whether patterns of endophyte diversity are correlated with host phylogeny or geography, we designed a broad geographic survey with controlled phylogenetic host sampling. We collected liverworts in North Carolina, Washington, Idaho, British Columbia, Germany, and New Zealand and identified endophytes using culture-based and molecular methods. Of the major lineages of filamentous ascomycetes recovered, 53-88% belonged to the Xylariales. Endophyte accumulation curves did not saturate, and singleton sequences were dominant in each region, suggesting that liverwort endophyte communities are diverse. There was no significant difference in species richness between regional endophyte communities; however, total richness estimators indicated that North Carolina and New Zealand have richer communities than do Germany and the Pacific Northwest. This pattern reflects lower per-host endophyte density and prevalence of a common, shared sequence group in Germany and the Pacific Northwest. Although species richness was relatively low in the Pacific Northwest, the greatest phylogenetic diversity of endophytes was recovered there. Tests for regional and host specificity revealed that endophyte floras of hosts within a geographic area are more similar to one another than to those of closely related hosts. Geographic distance, not host phylogeny, best explains differences among communities. PMID- 21632415 TI - Pollination efficiencies of flower-visiting insects as determined by direct genetic analysis of pollen origin. AB - The amount and genetic composition of pollen grains that are transported to flowers influence the reproduction and fitness of plants. Despite the importance of insect-pollination systems, an understanding of those systems is still lacking due to the absence of a genetic analysis of pollen grains that are transported to flowers. We evaluated the pollination efficiencies of bumblebees (Apidae, Bombus spp.), flower beetles (Scarabaeidae, subfamily Cetoniinae, Protaetia and Eucetonia sp.), and small beetles (Lagriidae, Arthromacra sp.) that visited the flowers of Magnolia obovata (Magnoliaceae) using quantitative (flower visitation frequency, amount of adherent pollen per insect) and qualitative (origin and genetic diversity of adherent pollen per insect) criteria. Most of the pollen adhering to bumblebees and small beetles was self-pollen. This result suggests that visitation by these insects may cause geitonogamous pollen flow and negatively affect the reproduction of M. obovata, causing inbreeding depression. In contrast, flower beetles transported large amounts of genetically diverse outcross pollen. Our results suggest that certain beetle species contribute quantitatively and qualitatively to the pollination of M. obovata. Direct genetic analysis of pollen grains will advance our understanding of plant mating systems and may shed light on the mutualism and coevolution of plants and flower visitors. PMID- 21632416 TI - Plasticity in salt tolerance traits allows for invasion of novel habitat by Japanese knotweed s. l. (Fallopia japonica and F.xbohemica, Polygonaceae). AB - Japanese knotweeds are among the most invasive organisms in the world. Their recent expansion into salt marsh habitat provides a unique opportunity to investigate how invasives establish in new environments. We used morphology, cytology, and AFLP genotyping to identify taxa and clonal diversity in roadside and salt marsh populations. We conducted a greenhouse study to determine the ability to tolerate salt and whether salt marsh populations are more salt tolerant than roadside populations as measured by the efficiency of PSII, leaf area, succulence, height, root-to-shoot ratio, and total biomass. Clonal diversity was extremely low with one F. japonica clone and five F. *bohemica genotypes. The two taxa were significantly different in several traits, but did not vary in biomass or plasticity of any trait. All traits were highly plastic in response to salinity, but differed significantly among genets. Despite this variation, plants from the salt marsh habitats did not perform better in the salt treatment, suggesting that they are not better adapted to tolerate salt. Instead, our data support the hypothesis that plasticity in salt tolerance traits may allow these taxa to live in saline habitats without specific adaptation to tolerate salt. PMID- 21632417 TI - Functional groups of rare plants differ in levels of imperilment. AB - Comparative examination of a large sample of plant species can reveal important aspects of life history that influence the ecology and distribution of taxa and their vulnerability to local extinction. We investigated whether functional groups of 71 rare plant species with contrasting life history traits differed in terms of population losses over time, regional range contraction, and range-wide levels of imperilment. Using town-level occurrence data from herbaria and Natural Heritage Program databases, we characterized species' extents of occurrence as alpha-hulls encompassing the centroids of New England towns that contained well documented populations of these rare taxa. Family affiliation was used as a covariate in analyses to reduce phylogenetic bias. Disparate functional groups of plants differed both in proportions of populations lost and declines in range areas over time, with insect-pollinated taxa, upland (vs. wetland) taxa, species with localized seed dispersal modes, and taxa reaching their northern range boundary in New England significantly more imperiled than other functional groups. These techniques permit a broad comparative assessment of the distribution of large numbers of plant taxa, so that we can identify several functional groups that warrant more concerted conservation attention. PMID- 21632418 TI - Menispermaceae from the Cerrejon Formation, middle to late Paleocene, Colombia. AB - The origin and processes creating the high diversity of plant species in neotropical rain forests and their floristic composition and multistratitified forest structure are still uncertain. Here, we studied one of the most common leaf morphotypes of the Cerrejon flora (middle-late Paleocene, ca. 60-58 Ma), Guajira, Colombia, that contains one of the oldest records of neotropical rain forest floras. Fifty-seven leaf specimens were carefully examined with a focus on general morphology, venation patterns, and cuticular characteristics. The analysis allowed us to recognize four new species that were assigned to the fossil-leaf genus Menispermites on the basis of an ovate leaf shape with cordate to truncate bases, actinodromous primary venation, brochidodromous secondary venation, percurrent tertiary venation, regular polygonal reticulate fourth and fifth venation, well-developed polygonal areoles, entire margin, and the presence of a fimbrial vein. This set of characters suggests a possible affinity with the pantropical angiosperm family Menispermaceae. The predominantly climbing habit of this family suggests that the Cerrejon Paleocene tropical rain forest was already multistratified. These findings represent the earliest record for the family in northern South America. PMID- 21632419 TI - Flower morphology and pollinator dynamics in Solanum carolinense (Solanaceae): implications for the evolution of andromonoecy. AB - Flower morphology and pollinator dynamics play an important role in the evolution and maintenance of many breeding systems, including andromonoecy. We used a series of field experiments to test the functional relationship between flower morphology and pollination dynamics (e.g., pollen receipt and export) in Solanum carolinense. We find that long-styled flowers serve primarily as pollen recipients and short-styled flowers as pollen donors, making this the first study to support the male-female interference hypothesis for the evolution of andromonoecy. However, this difference in the primary male or female function of the flowers depends on the pollinator identity. In flowers visited by Bombus impatiens, style length has a positive relationship with pollen deposition and a negative relationship with pollen removal. In contrast, neither morphological nor behavioral traits determine pollen deposition or removal by small halictid bees. We demonstrate that different pollinators could select for different floral morphologies, and thus, our research suggests that pollinator-specific interactions with flower morphology play an important role in the evolution and maintenance of anrdromonoecy. PMID- 21632420 TI - Phylogeny of Rutaceae based on twononcoding regions from cpDNA. AB - Primarily known only by the edible fruits of Citrus, Rutaceae comprise a large (c. 160 genera and 1900 species), morphologically diverse, cosmopolitan family. Of its extraordinary array of secondary chemical compounds, many have medicinal, antimicrobial, insecticidal, or herbicidal properties. To assist with the much needed suprageneric reclassification and with studies of evolution of chemical compounds and biogeographic history of the family, here we included sequence data (from two noncoding regions of the chloroplast genome-rps16 intron and trnL-trnF region) from 65 species in 59 genera (more than one third of those in the family) that represented all subfamilies and tribes and more genera of Toddalioideae and of neotropical groups than previous studies. Results confirmed that Cneorum, Ptaeroxylon, Spathelia, and Dictyoloma form a clade sister to the remaining Rutaceae, none of the subfamilies with more than one genus (except Aurantioideae) is monophyletic, and characters of the ovary and fruit are not reliable for circumscription of subfamilies. Furthermore, clades are better correlated with geographic distributions of the genera than with ovary and fruit characters. Circumscriptions of subfamilies and tribes (and some subtribes of Rutoideae) must be reevaluated. Results are discussed in light of geographic distributions, caryology, chemotaxonomy, and other molecular studies. PMID- 21632421 TI - Trans-Atlantic dispersal and large-scale lack of genetic structure in the circumpolar, arctic-alpine sedge Carex bigelowii s. l. (Cyperaceae). AB - Paradoxically, several of the ecologically most important plant groups in the Arctic are little understood in terms of taxonomy and biogeographic history. The circumpolar Carex bigelowii s. l. (Cyperaceae) is abundant in the Arctic and is one of the most complicated arctic plant groups. While its ecology and population genetics have been extensively studied, its taxonomy is largely unexplored. We analyzed the large-scale geographical structuring of amplified fragment length polymorphisms (AFLPs) covering most of the distribution range. We detected high levels of genetic variation, most (66%) within populations, and a fairly weak genetic structure. Only the Central Asian populations, referred to as C. orbicularis, were strongly divergent. For the remaining populations, Bayesian clustering separated three distinct clusters (one European, one amphi-Atlantic, and one broadly amphi-Beringian), probably reflecting different major glacial refugia and recent transoceanic dispersal. The isolated central European populations were most closely related to those from a larger distribution area in northern Europe. Differences in genetic diversity suggest that the Alpine and Tatra populations have experienced strong bottlenecks, whereas the Krkonose population may have been part of a continuous distribution area during the cold stages of the Pleistocene. Finally, we discuss the relevance of our results for a uniform, range-wide taxonomic concept. PMID- 21632422 TI - Evolutionary relationships in the showy mistletoe family (Loranthaceae). AB - Loranthaceae (73 genera and ca. 900 species) comprise mostly aerial hemiparasitic plants. Three monotypic genera considered relicts are root parasites. The family is diverse in tropical areas, but representatives are also found in temperate habitats. Previous classifications were based on floral and inflorescence morphology, karyological information, and biogeography. The family has been divided into three tribes: Nuytsiae, Elytrantheae (subtribes Elytranthinae and Gaiadendrinae), and Lorantheae (subtribes Loranthinae and Psittacanthinae). Nuytsiae and Elytrantheae are characterized by a base chromosome number of x = 12, whereas subtribes Loranthinae (x = 9) and Psittacanthinae (x = 8) numbers are derived via aneuploid reduction. To elucidate the phylogeny of the family, we analyzed sequences from five genes (nuclear small and large subunit rDNA and the chloroplast genes rbcL, matK, and trnL-F) representing most genera using parsimony, likelihood, and Bayesian inference. The three root parasites, Nuytsia, Atkinsonia, and Gaiadendron, are supported as successive sister taxa to the remaining genera, resulting in a monophyletic group of aerial parasites. Three major clades are resolved each corresponding to a subtribe. However, two South American genera (Tristerix and Notanthera) and the New Zealand genus Tupeia, which were previously classified in subtribe Elytranthinae, are weakly supported as part of a clade representing the South American subtribe Psittacanthinae. PMID- 21632423 TI - Phylogeny, biogeography, and rates of diversification of New World Astragalus (Leguminosae) with an emphasis on South American radiations. AB - This study uses phylogenetic relationships of New World representatives of the species-rich genus Astragalus (Leguminosae; Papilionoideae) to follow up on recent evidence pointing to rapid and recent plant diversification patterns in the Andes. Bayesian and maximum likelihood phylogenetic analyses were done using nuclear rDNA ITS and chloroplast spacers trnD-trnT and trnfM-trnS1, either separately or in combination. The effect of using partitioned vs. nonpartitioned analyses in a Bayesian approach was evaluated. Highest resolution was obtained when the data were combined in partitioned or nonpartitioned Bayesian analyses. All phylogenies support two clades of South American species nested within the North American species, implying two separate invasions from North to South America. These two clades correspond to the original morphological classification of Johnston (1947 Journal of the Arnold Arboretum 28: 336-409). The mean ages of the South American clades were very recent but still significantly different (1.89 and 0.98 Ma). Upper and lower bounds on rates of diversification varied between 2.01 and 0.65 species/Ma for the older clade and 2.06 and 1.24 species/Ma for the younger clade. Even the lower bounds are still very high, reasserting Neo Astragalus in the growing list of recent rapid radiations of plants, especially in areas with a high physiographic diversity, such as the Andes. PMID- 21632424 TI - Neotropical roots of a Polynesian spice: the hybrid origin of Tahitian vanilla, Vanilla tahitensis (Orchidaceae). AB - Absent in the wild, Tahitian vanilla (V. tahitensis) is a gourmet spice restricted in distribution to cultivated and feral stands in French Polynesia and Papua New Guinea. Its origins have been elusive. Our objective was to test the purported hybrid derivation and parentage of V. tahitensis from aromatic, neotropical progenitors. Nucleotide sequences from V. tahitensis and neotropical Vanilla were assayed for phylogenetic relatedness in two independently inherited genomic regions, the nuclear ITS region, and the trnH-psbA noncoding region of chloroplast DNA. As predicted to occur for early generation hybrids, placement of V. tahitensis was nonconcordant. All V. tahitensis clustered with V. planifolia from analysis of cpDNA sequences, suggesting V. planifolia as the maternal genome contributor. Phylogenetic reconstruction of ITS sequences showed that most V. tahitensis nested incongruently with V. odorata, but others remained sister to V. planifolia. Recovery of ITS clones in V. tahitensis related to both V. planifolia and V. odorata also supports its biphyletic origin from these two taxa. We interpret the high percentage (95%) of additive polymorphic sites in V. tahitensis relative to its parents as indication of a recent, and probably human mediated, evolutionary origin. PMID- 21632425 TI - KDEL-tailed cysteine endopeptidases involved in programmed cell death, intercalation of new cells, and dismantling of extensin scaffolds. AB - KDEL-tailed cysteine endopeptidases are a group of papain-type peptidases found in senescing tissue undergoing programmed cell death (PCD). Their genes have so far been cloned and analyzed in 12 angiosperms. They are synthesized as proenzymes with a C-terminal KDEL endoplasmatic reticulum retention signal, which is removed with the prosequence to activate enzyme activity. We previously identified three genes for KDEL-tailed cysteine endopeptidases (AtCEP1, AtCEP2, AtCEP3) in Arabidopsis thaliana. Transgenic plants of A. thaliana expressing beta glucuronidase (GUS) under the control of the promoters for the three genes were produced and analyzed histochemically. GUS activity was promoter- and tissue specific GUS activity during seedling, flower, and root development, especially in tissues that collapse during final stages of PCD, and in the course of lateral root formation. KDEL-tailed cysteine endopeptidases are unique in being able to digest the extensins that form the basic scaffold for cell wall formation. The broad substrate specificity is due to the structure of the active site cleft of the KDEL-tailed cysteine endopeptidase that accepts a wide variety of amino acids, including proline and glycosylated hydroxyproline of the hydroxyproline rich glycoproteins of the cell wall. PMID- 21632426 TI - Host genotype overrides fungal endophyte infection in influencing tiller and spike production of Lolium perenne (Poaceae) in a common garden experiment. AB - Leaves of many cool-season grasses are infected by endophytic fungi that can impact their populations. A common garden experiment with Lolium perenne was established in a lawn in New Jersey, USA, to investigate the impact of endophyte infection and host genotype on tiller and spike production over three years. Infected (E+) and uninfected (E-) plants of each genotype were monitored every 2 3 mo. Infection intensity within plants varied significantly among genotypes and years, but there was no evidence of directional change over time. Tiller production varied significantly among genotypes and was affected by endophytes: E+ plants of several genotypes produced more tillers than E- plants during the third year. E+ plants had greater aboveground biomass, but host genotype explained a far greater proportion of variation in tiller production, number, and biomass than infection. Plant survival, percentage flowering, flowering date, number of spikes, and mean tiller mass were unaffected by endophytes. However, the last three variables showed significant variation among host genotypes. Although studies have demonstrated a positive growth effect of endophytes on several grass hosts, in this experiment host genotype accounted for far more of the variation in tiller and spike production and in biomass of Lolium perenne than endophyte infection. PMID- 21632427 TI - Pollinator and nonpollinator selection on ray morphology in Leucanthemum vulgare (oxeye daisy, Asteraceae). AB - Despite evidence that both pollinators and nonpollinator agents of selection can shape the evolution of floral characters, there have been few attempts to compare the strengths and directions of selection from pollinators and other agents in the same study system. In this investigation of Leucanthemum vulgare, a self incompatible composite known for its conspicuous white rays, I obtained data from a ray removal experiment in the field and from a segregating F(2) population in an experimental garden to assess the role of pollinator and nonpollinator selection as stabilizing factors on floral evolution in this species. Removal of all rays reduced the pollination success of heads by 31-35%, but did not significantly affect the level of infestation by larvae of the fly Tephritis neesii. Data from F(2) plants indicated a potential for indirect selection on ray morphology, mediated through links between ray morphology and measures of vegetative size and plant vigor. The results of this study show that individuals of the normal, rayed phenotype have a clear selective advantage, in terms of both pollinator attraction and general plant vigor. Thus, there were no conflicting selection pressures between the pollinators and the other selective agents considered in this study. PMID- 21632428 TI - In search of monophyletic taxa in the family Desmidiaceae (Zygnematophyceae, Viridiplantae): the genus Cosmarium. AB - Nuclear-encoded small subunit (SSU) rDNA, 1506 group I introns, and chloroplast rbcL genes were sequenced from 97 strains representing the largest desmid genus Cosmarium (45 spp.), its putative relatives Actinotaenium (5 spp.), Xanthidium (4 spp.), Euastrum (9 spp.), Staurodesmus (13 spp.), and other Desmidiaceae (Zygnematophyceae, Streptophyta) and used to assess phylogenetic relationships in the family. Analyses of single genes and of a concatenated data set (3260 nt) established 10 well-supported clades in the family with Cosmarium species distributed in six clades and one nonsupported assemblage. Most of the clades contained representatives of at least two genera highlighting the polyphyletic nature of the genera Cosmarium, Euastrum, Staurodesmus, and Actinotaenium. To enhance resolution between clades, we extended the data set by sequencing the slowly evolving chloroplast-encoded large subunit (LSU) rRNA gene from 40 taxa. Phylogenetic analyses of a concatenated data set (5509 nt) suggested a sister relationship between two clades that consisted mainly of Cosmarium species and included C. undulatum, the type species of the genus. We describe molecular signatures in the SSU rRNA for two clades and conclude that more studies involving new isolates, additional molecular markers, and reanalyses of morphological traits are necessary before the taxonomic revision of the genus Cosmarium can be attempted. PMID- 21632429 TI - Hidden diversity of endophytic fungi in an invasive plant. AB - Fungal endophytes are important in plant ecology and common in plants. We attempted to test cointroduction and host-jumping hypotheses on a community basis by comparing endophytes isolated from invasive spotted knapweed (Centaurea stoebe, Asteraceae) in its native and invaded ranges. Of 92 combined, sequence based haplotypes representing eight classes of Fungi, 78 occurred in only one of the two ranges. In the native range of C. stoebe, one haplotype of Alternaria alternata was clearly dominant, whereas in the invaded range, no haplotype was dominant. Many haplotypes were closely related to one another and novel. For example, six putative, new species of Botrytis were discovered as endophytes of C. stoebe, which has never been reported to have Botrytis spp.. Apparent differences between the two communities of endophytes were significant according to an analysis of similarity, but phylogenetic community structure did not differ significantly between the ranges. Both host-jumping and cointroduction of fungal endophytes likely took place during the spotted knapweed invasion. PMID- 21632430 TI - Hybridization and crossability in Caiophora (Loasaceae subfam. Loasoideae): Are interfertile species and inbred populations results of a recent radiation? AB - Interspecific hybridization is considered a possible mechanism of plant diversification. The Andes are a hotspot of biodiversity, but hybridization in Andean taxa has so far not been investigated intensively. The current study investigates crossability in Caiophora (Loasaceae subfam. Loasoideae) by experimental interspecific hybridization of seven different species. Hand pollination was undertaken, developing fruits counted, thousand (seed) grain weights, and seed viability were examined. Cross pollination led to some fruit set in 36 of the 37 different combinations. Overall fruit set was virtually identical irrespective of the combination of parental plants. Mean germination rates were much higher in hybrid seeds, indicating a marked heterosis effect and the possible presence of an inbreeding depression in the source populations: In experimental hybridization the divergent taxa of Caiophora behave like isolated, inbred populations of a single species. Allopatry and different habitat preferences seem to be the key factors keeping the (interfertile) taxa of Caiophora apart in the apparent absence of both postmating isolating mechanisms and obvious isolating mechanisms in phenology and floral biology. Interspecific hybrids reported from the wild appear to be the result of secondary contact due to human impact. PMID- 21632431 TI - Incongruence between cpDNA and nrITS trees indicates extensive hybridization within Eupersicaria (Polygonaceae). AB - Interspecific hybridization followed by polyploidization appears to have played a major role in plant diversification, but quantifying the contribution of this mechanism to diversification within taxonomically complex clades remains difficult. Incongruence among gene trees can provide critical insights, especially when combined with data on chromosome numbers, morphology, and geography. To further test our previous hypothesis on hybrid speciation in Persicaria (Polygonaceae), we performed molecular phylogenetic studies using three cpDNA regions and nuclear ITS sequences, with an emphasis on sampling within section Eupersicaria. Our analyses revealed major conflicts between the combined cpDNA tree and the nrITS tree; a variety of incongruence tests rejected stochastic error as the cause of incongruence in most cases. On the basis of our tree incongruence results and information on chromosome numbers, we hypothesize that the origin of 10 polyploid species involved interspecific hybridization. Our studies also support the recognition of several previously named species that have been treated as belonging within other species. Repeated allotetraploidy (as distinct from radiation at the tetraploid level) now appears to be the key mechanism governing the diversification of this taxonomically challenging group. PMID- 21632432 TI - Toward a comprehensive understanding of phylogenetic relationships among lineages of Acanthaceae s.l. (Lamiales). AB - Acanthaceae (Asteridae; Lamiales) include ~4000 species and encompass a range of morphological diversity, habitats, and biogeographic patterns. Although they are important components of tropical and subtropical habitats worldwide, inadequate knowledge of the family's phylogenetic framework has impeded comparative research. In this study, we sampled all known lineages of Acanthaceae including Andrographideae. Also included were eight of 13 genera whose relationships remain enigmatic. We used sequence data from nrITS and four chloroplast noncoding regions, and parsimony and Bayesian methods of analysis. Results strongly support most aspects of relationships including inclusion of Avicennia in Acanthaceae. Excepting Neuracanthus, newly sampled taxa are placed with strong support; Kudoacanthus is in Justicieae, Tetramerium lineage, and the remaining enigmatic genera are in Whitfieldieae or Barlerieae, and Andrographideae are sister to Barlerieae. This last result is unanticipated, but placement of Andrographideae based on structural characters has been elusive. Neuracanthus is monophyletic but placement relative to (Whitfieldieae (Andrographideae + Barlerieae)) is weakly supported. Many clades have clear morphological synapomorphies, but nonmolecular evidence for some remains elusive. Results suggest an Old World origin with multiple dispersal events to the New World. This study informs future work by clarifying sampling strategy and identifying aspects of relationships that require further study. PMID- 21632433 TI - Phylogenetic relationships of aroids and duckweeds (Araceae) inferred from coding and noncoding plastid DNA. AB - Familial, subfamilial, and tribal monophyly and relationships of aroids and duckweeds were assessed by parsimony and Bayesian phylogenetic analyses of five regions of coding (rbcL, matK) and noncoding plastid DNA (partial trnK intron, trnL intron, trnL-trnF spacer) for exemplars of nearly all aroid and duckweed genera. Our analyses confirm the position of Lemna and its allies (formerly Lemnaceae) within Araceae as the well-supported sister group of all aroids except Gymnostachydoideae and Orontioideae. The last two subfamilies form the sister clade of the rest of the family. Monophyly of subfamilies Orontioideae, Pothoideae, Monsteroideae, and Lasioideae is supported, but Aroideae are paraphyletic if Calla is maintained in its own subfamily (Calloideae). Our results suggest expansion of the recently proposed subfamily Zamioculcadoideae (Zamioculcas, Gonatopus) to include Stylochaeton and identify problems in the current delimitation of tribes Anadendreae, Heteropsideae, and Monstereae (Monsteroideae), Caladieae/Zomicarpeae, and Colocasieae (Aroideae). Canalization of traits of the spathe and spadix considered typical of Araceae evolved after the split of Gymnostachydoideae, Orontioideae, and Lemnoideae. An association with aquatic habitats is a plesiomorphic attribute in Araceae, occurring in the helophytic Orontioideae and free-floating Lemnoideae, but evolving independently in various derived aroid lineages including free-floating Pistia (Aroideae). PMID- 21632434 TI - Evolution and phylogenetic utility of the PHOT gene duplicates in the Verbena complex (Verbenaceae): dramatic intron size variation and footprint of ancestral recombination. AB - A well-resolved species level phylogeny is critically important in studying organismal evolution (e.g., hybridization, polyploidization, adaptive speciation). Lack of appropriate molecular markers that give sufficient resolution to gene trees is one of the major impediments to inferring species level phylogenies. In addition, sampling multiple independent loci is essential to overcome the lineage sorting problem. The availability of nuclear loci has often been a limiting factor in plant species-level phylogenetic studies. Here the two PHOT loci were developed as new sources of nuclear gene trees. The PHOT1 and PHOT2 gene trees of the Verbena complex (Verbenaceae) are well resolved and have good clade support. These gene trees are consistent with each other and previously generated chloroplast and nuclear waxy gene trees in most of the phylogenetic backbone as well as some terminal relationships, but are incongruent in some other relationships. Locus-specific primers were optimized for amplifying and sequencing these two loci in all Lamiales. Comparing intron size in the context of the gene trees shows dramatic variation within the Verbena complex, particularly at the PHOT1 locus. These variations are largely caused by invasions of short transposable elements and frequent long deletions and insertions of unknown causes. In addition, inspection of DNA sequences and phylogenetic analyses unmask a clear footprint of ancestral recombination in one species. PMID- 21632435 TI - Molecular phylogeny and adaptive radiation of the endemic Hawaiian Plantago species (Plantaginaceae). AB - Insular oceanic islands provide excellent opportunities for the study of evolutionary processes and adaptive radiation. The Hawaiian Plantago radiation comprises six endemic taxa showing considerable inter- and intraspecific morphological and ecological diversity. The rDNA internal (ITS) and external (ETS) transcribed spacers and two recently described chloroplast spacers, ndhF rpl32 and rpl32-trnL, were sequenced to study phylogenetic relationships within this morphologically complex group. Phylogenetic analysis provided strong evidence for the monophyly of Hawaiian Plantago, suggesting that the lineage arose from a single long-distance dispersal event. Inconsistencies between nuclear and chloroplast phylogenies suggest a history of hybridization. The basal, unresolved dichotomy of the combined phylogeny is consistent with rapid phenotypic diversification of the major lineages early in the history of this group. Speciation has largely occurred allopatrically, with divergence a result of intraisland ecological shifts between bog and woodland habitats and interisland dispersal events. Most interisland colonizations were from older to younger islands with initial colonization of Kaua'i. In our analysis, P. pachyphylla is paraphyletic and taxonomic separation of the distinct morphotypes of this species appears justified. Furthermore, the apparent hybrid ancestry and unique morphology and habitat of the endangered P. princeps var. longibracteata support its recognition at the specific rank. PMID- 21632436 TI - Male gender role strain as a barrier to African American men's physical activity. AB - Despite the potential health consequences, African American men tend to treat their roles as providers, fathers, spouses, and community members as more important than engaging in health behaviors such as physical activity. We conducted 14 exploratory focus groups with 105 urban, middle-aged African American men from the Midwest to examine factors that influence their health behaviors. Thematic content analysis revealed three interrelated barriers to physical activity: (a) work, family, and community commitments and priorities limited time and motivation for engaging in physical activity; (b) physical activity was not a normative individual or social activity and contributed to men prioritizing work and family responsibilities over physical activity; and (c) the effort men exerted in seeking to fulfill the provider role limited their motivation and energy to engage in physical activity. These findings highlight the need for physical activity interventions that consider how health fits in the overall context of men's lives. PMID- 21632437 TI - Correlates of adherence to a telephone-based multiple health behavior change cancer preventive intervention for teens: the Healthy for Life Program (HELP). AB - This study examined factors associated with teens' adherence to a multiple health behavior cancer preventive intervention. Analyses identified predictors of trial enrollment, run-in completion, and adherence (intervention initiation, number of sessions completed). Of 104 teens screened, 73% (n = 76) were trial eligible. White teens were more likely to enroll than non-Whites (chi(2)[1] df = 4.49, p = .04). Among enrolled teens, 76% (n = 50) completed the run-in; there were no differences between run-in completers and noncompleters. A majority of run-in completers (70%, n = 35) initiated the intervention, though teens who initiated the intervention were significantly younger than those who did not (p < .05). The mean number of sessions completed was 5.7 (SD = 2.6; maximum = 8). After adjusting for age, teens with poorer session engagement (e.g., less cooperative) completed fewer sessions (B = -1.97, p = .003, R (2) = .24). Implications for adolescent cancer prevention research are discussed. PMID- 21632438 TI - The effect of distressing imagery on attention to and persuasiveness of an antialcohol message: a gaze-tracking approach. AB - BACKGROUND: Distressing imagery may inhibit health communications by inducing audiences to reduce distress by avoiding attention to persuasive messages. METHOD: This study used eye-tracking methods to compare gaze time allocated to a persuasive textual message, accompanied by either distressing high-resolution color images or less distressing two-color images with degraded outline and detail. RESULTS: Participants in the distressing images condition showed lower intentions to reduce drinking in the following 3 months, which may have been mediated by lower gaze time to textual elements of the message. The effect was stronger in participants who both scored lower on dispositional mental disengagement and were more vulnerable to alcohol-related problems. CONCLUSIONS: These findings suggest that distressing imagery may inhibit persuasion by reducing audience attention to message components. Implications for message design are discussed. PMID- 21632439 TI - Gender and health lifestyle: an in-depth exploration of self-care activities in later life. AB - OBJECTIVE. Evaluate similarities and differences in the self-care domain of health lifestyle among older, rural-dwelling women and men. METHOD: Qualitative analysis of in-depth interview data from 62 community-dwelling older (M = 74.3 years) African and European American women and men. RESULTS: Both older women and men rely heavily on over-the-counter medications and home remedies for self-care; professional health care is typically sought when self-care is not effective. However, relative to men, women were more knowledgeable about different approaches to self-care, especially home remedies; they used a wider range of self-care activities; and they placed greater priority on self-care over professional health care. DISCUSSION: The structure of older women's and men's self-care domain of health lifestyle is similar. However, there are subtle differences in health lifestyle that are likely embedded in gendered role behavior and may contribute to women's greater health complaints. PMID- 21632440 TI - Compared with younger peritoneal dialysis patients, elderly patients have similar peritonitis-free survival and lower risk of technique failure, but higher risk of peritonitis-related mortality. AB - BACKGROUND: The number of elderly patients with end-stage kidney disease (ESKD) is increasing worldwide, but the proportion of elderly patients commencing peritoneal dialysis (PD) is falling. The reluctance of elderly ESKD patients to consider PD may be related to a perception that PD is associated with greater rates of complications. In the present study, we compared outcomes between younger and older PD patients. METHODS: Using Australia and New Zealand Dialysis Registry data, all adult ESKD patients commencing PD between 1991 and 2007 were categorized into under 50, 50 - 64.9, and 65 years of age or older groups. Time to first peritonitis, death-censored technique failure, and peritonitis associated and all-cause mortality were evaluated by multivariate Cox proportional hazards model analysis. RESULTS: Of the 12 932 PD patients included in the study, 3370 (26%) were under 50 years of age, 4386 (34%) were 50 - 64.9 years of age, and 5176 (40%) were 65 years of age or older. Compared with younger patients (<50 years), elderly patients (>= 65 years) had a similar peritonitis free survival and a lower risk of death-censored technique failure [hazard ratio (HR): 0.85; 95% confidence interval (CI): 0.79 to 0.93], but they had higher peritonitis-related (HR: 2.31; 95% CI: 1.68 to 3.18) and all-cause mortality (HR: 2.90; 95% CI: 2.60 to 3.23). CONCLUSIONS: Not unexpectedly, elderly patients have higher peritonitis-related and all-cause mortality, which is likely a consequence of a greater prevalence of comorbid disease. However, compared with younger patients, elderly patients have superior technique survival and similar peritonitis-free survival, suggesting that PD is a viable renal replacement therapy in this group of patients. PMID- 21632441 TI - Use of peritoneal dialysis after surgery for congenital heart disease in children. AB - Acute kidney injury (AKI) is a common complication in children after surgery for congenital heart disease, and peritoneal dialysis (PD) is usually the renal replacement therapy (RRT) of choice, especially in very young children. The aim of the present study was to describe our experience of using PD to treat AKI after cardiac surgery. We retrospectively analyzed children 1 week to 16 years of age undergoing cardiac surgery during 2000-2008 and found the incidence of AKI treated with PD to be 2.3%. In the 23 patients treated with PD (13 male; average age: 29 +/- 48.4 months; weight: 9.1 +/- 8.1 kg), the indications for PD initiation were oliguria (n = 13), anuria (n = 9), and acidosis (n = 1). The average time between cardiac surgery and AKI was 4.8 +/- 16.8 hours, and between AKI and PD initiation, it was 12 +/- 16.8 hours. Patients were treated for a mean of 4.8 +/- 3.8 days. Two patients developed peritonitis, and mechanical dysfunction of the PD catheter occurred in 1 patient. In-hospital mortality was 43.4%. Patients treated with PD weighed less (p = 0.004) and had longer bypass time (p = 0.004), inotrope use (p = 0.000), and mechanical ventilation (p = 0.000). However, in a regression analysis, only cardiopulmonary bypass time (odds ratio: 1.021; 95% confidence interval: 0.998 to 1.027; p = 0.032) remained predictive of a subsequent need for PD. We conclude that PD is an efficacious RRT for AKI in children undergoing cardiac surgery and that, in this setting, bypass time is the strongest predictor of a subsequent need for RRT. PMID- 21632442 TI - Assessment of oral health in peritoneal dialysis patients with and without diabetes mellitus. AB - The incidence of chronic renal failure continues to rise worldwide, and although the oral and dental changes in individuals with this condition have been examined, investigations with diabetic peritoneal dialysis (PD) patients are limited. We therefore examined salivary pH, dry mouth, taste change, and mucosal ulceration in diabetic and nondiabetic uremic patients receiving PD. A total of 49 patients undergoing PD therapy were allocated to either the diabetic or the nondiabetic group. Salivary pH, dry mouth, taste change, and mucosal ulceration were determined for both groups. Salivary flow rate and pH were both lower in the diabetic group. Buffer capacity, dry mouth, taste change, and mucosal ulceration were all increased in that group. These findings were associated with level of glycosylated hemoglobin A1c. Our observations indicate that, compared with nondiabetic PD patients, patients with diabetes exhibit more severe oral uremic symptoms, including dry mouth, burning mouth syndrome, taste change, and mucosal ulcerations. The oral health in these patients should be followed. PMID- 21632444 TI - Prolonged duration of peritoneal dialysis catheter embedment does not lower the catheter success rate. AB - OBJECTIVE: Since 2000, we have used the Moncrief-Popovich technique as our standard method for peritoneal dialysis (PD) catheter insertion. The subcutaneous portion of the catheter is externalized immediately before initiation of PD. We undertook the present review to investigate whether duration of catheter embedment affects catheter or patient outcome. METHODS: All catheters inserted beginning 1 January 2000 and externalized by 31 December 2008 were included. The primary outcome was catheter survival. Secondary outcomes were catheter patency (no fibrin plug or omental wrap) and complications within 90 days after externalization. A standard peritoneal equilibration test was used to classify peritoneal membrane transport status. Proportional hazards regression models were used to test whether duration of embedment affected catheter outcomes. The models treated embedment duration as both a continuous predictor and a categorical predictor categorized by tertile. RESULTS: A total of 134 catheters were implanted and externalized. Twelve patients received 2 catheters each. To ensure statistical independence of the observations, 12 of the latter 24 catheters were excluded (1 chosen randomly from each patient), resulting in a useable sample size of 122 catheters. The total duration of observation was 2359 patient-months. The median duration of catheter embedment was 40.5 days (range: 2 - 788 days). After controlling for sex, race, age, and diabetes status, embedment duration did not have a significant effect on catheter survival as a continuous predictor or as a categorical predictor. Additionally, the 95% confidence interval for the 30 day effect of embedment duration ruled out a change of more than 20.6% in the hazard of catheter malfunction or infection. Of the studied catheters, 89.3% were patent and functioned properly immediately upon externalization. The remaining 13 catheters (10.7%) lacked patency on externalization because of fibrin plug or kinking (n = 10) or omental wrap (n = 3); however, 12 of the 13 non-patent catheters were corrected laparoscopically, and the patients resumed PD. Only 1 patient transferred to hemodialysis. Overall, 121 of 122 buried catheters (99.2%) were used for PD. Other complications within 90 days of catheter externalization included incision site and tunnel infection in 2 cases (1.6%), exit-site leak in 2 cases (1.6%), and coagulase-negative staphylococcal peritonitis in 1 case (0.8%). CONCLUSIONS: Duration of catheter embedment before externalization did not affect catheter survival and did not influence subsequent peritoneal membrane transport status. The overall effect of increasing embedment duration by 30 days is, at most, a 20.6% increase or decrease in the hazard of catheter failure, but the actual hazard may be much smaller or nonexistent. Larger studies are needed to further explore the ideal duration of embedment. PMID- 21632443 TI - "NEPP" peritoneal dialysis regimen has beneficial effects on plasma CEL and 3-DG, but not pentosidine, CML, and MGO. AB - BACKGROUND: Standard peritoneal dialysis (PD) solutions contain high levels of glucose and glucose degradation products (GDPs), both contributing to the formation of advanced glycation end products (AGEs). We studied the contribution to plasma GDP and AGE levels of 2 PD regimens that differ in glucose and GDP loads: high load [standard PD (sPD) using 4 glucose-lactate exchanges] and low load [1 amino acid exchange, 1 icodextrin exchange, and 2 glucose bicarbonate/lactate exchanges ("NEPP")]. METHODS: In a prospective crossover study (2 periods of 24 weeks), new continuous ambulatory PD patients were randomized to NEPP-sPD (n = 23) or to sPD-NEPP (n = 27). RESULTS: After the start of PD, absolute increases were observed in plasma levels of 3-deoxyglucosone (3 DG, 220.4 nmol/L, p < 0.0001) and in N(epsilon)-(carboxymethyl) lysine (CML) in plasma proteins (0.02 MUmol/L CML per 1 mol/L lysine, p < 0.0001). During the first 6 weeks, 3-DG tended to increase more with sPD treatment (p = 0.08), and CML, with NEPP treatment (p = 0.002). In both groups, N(epsilon) (carboxyethyl)lysine (CEL) in plasma proteins declined significantly with the start of PD. Treatment with NEPP resulted in higher levels of methylglyoxal (MGO) and lower levels of 3-DG and CEL. Pentosidine in the albumin fraction tended to increase less during NEPP treatment. CONCLUSIONS: A low glucose and GDP PD regimen (NEPP) resulted in plasma levels of 3-DG and CEL that were lower than those with a glucose-based sPD regimen. Starting PD with NEPP was associated with a steeper increase in CML, and continuing treatment with NEPP resulted in higher MGO levels. PMID- 21632445 TI - Three-pore model predictions of 24-hour automated peritoneal dialysis therapy using bimodal solutions. AB - BACKGROUND: Recently, bimodal peritoneal dialysis (PD) solutions containing low concentrations of Na have been shown to increase 24-hour ultrafiltration (UF) or UF efficiency (UF volume per gram of carbohydrate or CHO absorbed) and Na removal in high ("fast") transport patients during automated PD therapy. We used computer simulations to compare UF efficiency and Na removal at equivalent 24-hour UF volumes using either a generic bimodal solution (2.27% glucose + 7.5% icodextrin) during the long dwell or an increase in the glucose concentration during the short dwells, with all solutions containing Na at the conventional concentration (132 mEq/L). METHODS: The 3-pore model has been shown to accurately predict peritoneal transport for PD solutions containing glucose or icodextrin, or both. Here, we used that model to calculate 24-hour UF volume, CHO absorption, and Na removal for high (H), high-average (HA), and low-average (LA) transport patients on automated PD. Nighttime therapy consisted of 1.36% or 2.27% glucose solution (or both), and daytime therapy consisted of either Extraneal (Baxter Healthcare Corporation, Deerfield, IL, USA) or a bimodal solution. RESULTS: As expected, addition of glucose to either the long dwell or the short dwells resulted in increased UF volume and glucose absorption. The increase in UF was a function of patient transport type (bimodal range: 288 - 490 mL; short-dwell range: 323 - 350 mL), and the increase in CHO absorption was smaller with glucose added to short dwells than with bimodal solution (range: 18 - 30 g vs. 34 - 39 g). The 24-hour UF efficiency was higher when high glucose concentrations were used during short dwell exchanges than when a bimodal PD solution was used for the long dwell (0.6 to 1.2 mL/g vs. -0.1 to 0.5 mL/g). By contrast, Na removal was lower with the short-dwell exchanges (28.3 - 30.7 mmol vs. 36.2 - 53.3 mmol), likely because of more pronounced Na sieving. CONCLUSIONS: Our modeling studies predict that generic bimodal PD solutions will provide higher Na removal but not higher 24 hour UF efficiency compared with current automated PD prescriptions using Extraneal for the long dwell and glucose-containing solutions for the short dwells. The modeling predictions from this study require clinical validation. PMID- 21632446 TI - The fluid study protocol: a randomized controlled study on the effects of bioimpedance analysis and vitamin D on left ventricular mass in peritoneal dialysis patients. AB - OBJECTIVES: We will evaluate the effects of bioimpedance analysis-guided fluid management to reduce volume expansion, of vitamin D(3) supplementation, and of the combination of those techniques on decrease of left ventricular mass in peritoneal dialysis (PD) patients. DESIGN: This multicenter randomized controlled trial, with a 2 * 2 factorial design, will be conducted at PD clinics affiliated with 3 Canadian teaching hospitals. Consenting PD patients 18 years of age or older will be included. Patients will be excluded if they have contraindications to bioimpedance or magnetic resonance imaging, life or technique expectancy of less than 1 year, peritonitis within the preceding 3 months, or serum calcium above 2.55 mmol/L. INTERVENTION: The study will randomize 70 patients to bio impedance-guided volume management or to usual care and to vitamin D(3) 50,000 U weekly for 8 doses, and then 10,000 U weekly or to matching placebo. MAIN OUTCOME MEASURES: The primary outcome will be change in left ventricular mass at 1 year as determined by cardiac magnetic resonance imaging. The secondary outcome will be a composite endpoint of death, nonfatal cardiovascular event, and transfer to hemodialysis for dialysis inadequacy or ultrafiltration failure. Other outcome measures will include blood pressure, quality of life, 6-minute walk test, inflammatory and fibrotic markers and their association with peritoneal membrane transport properties, and residual renal function. Patients will be followed for clinical outcomes for up to 3 years. CONCLUSIONS: This study will assess whether bioimpedance-directed volume management and vitamin D(3) supplementation can improve left ventricular mass in PD patients. PMID- 21632447 TI - Experiences with assisted peritoneal dialysis in China. AB - OBJECTIVE: About half the patients on peritoneal dialysis (PD) in China need to be assisted by family members or home assistants. We explored whether these patients have a higher risk for peritonitis and death compared with self-care PD patients. METHODS: We prospectively followed 313 incident PD patients until death or censoring. This cohort was divided into assisted and self-care PD groups according to the independence of bag exchange. Data on baseline demographics, Charlson comorbidity index, biochemistry, and residual renal function were recorded during the first 3 - 6 months. The outcome variables were first episode of peritonitis and all-cause mortality. RESULTS: Of the 313 patients in this cohort study, 122 needed assistance in performing bag exchanges (86 from a family member, 36 from a home assistant); the remaining 191 patients did not need assistance. During a follow-up period averaging 44.5 months, 122 patients developed a first episode of peritonitis, and 135 patients died. Compared with patients having a family assistant, those with a home assistant had similar peritonitis-free and survival times, but a higher risk of mortality after adjustments for variables such as age, sex, Charlson comorbidity score, hemoglobin, serum albumin, and residual renal function. Furthermore, compared with self-care patients, assisted patients overall had a similar peritonitis-free time, but a higher risk of mortality, even after adjusting for covariates. CONCLUSIONS: Based on our single-center experience in China, we conclude that assisted PD is a good option for patients with poor self-care ability. This result provides evidence for recruiting patients who need assistance to PD programs in China. PMID- 21632448 TI - Overweight, obesity, diabetes, and risk of breast cancer: interlocking pieces of the puzzle. AB - We considered epidemiological data on overweight, diabetes, insulin, and breast cancer. Overweight is inversely related to premenopausal breast cancer, but there is definite evidence that, as compared with normal weight women, the relative risk (RR) for postmenopausal breast cancer is around 1.5 for overweight women and >2 for obese women, and that the association is stronger in elderly women. Overweight and obesity are strongly related to diabetes. Diabetes is associated with postmenopausal breast cancer, too, with summary RRs from meta-analyses of 1.15-1.20, but not with premenopausal breast cancer (RR, 0.9). There is no consistent evidence that fasting insulin is related to breast cancer risk. Thus, although overweight and obesity are strongly related to postmenopausal breast cancer, diabetes is only moderately related to it. Given the extent of the association, and the likely residual confounding by overweight, inference on causality for the diabetes-breast cancer relation remains open to discussion. PMID- 21632449 TI - Early clinical development of ARQ 197, a selective, non-ATP-competitive inhibitor targeting MET tyrosine kinase for the treatment of advanced cancers. AB - Expression of the receptor tyrosine kinase c-MET (MET, mesenchymal-epithelial transition factor) in many cancers, and its participation in multiple signal transduction pathways involved in malignant tumor growth, suggest a wide therapeutic potential for MET inhibition in human cancer. Here we describe the discovery and early clinical development of ARQ 197, a novel, selective, non-ATP competitive inhibitor of MET. Phase I studies demonstrate that ARQ 197 has a predictable pharmacokinetics and favorable safety profile, making it a potentially ideal partner for combination with cytotoxic chemotherapies and targeted anticancer agents. Results from phase I and phase II trials demonstrate preliminary evidence of anticancer activity. New data from a global phase II randomized trial comparing a combination of ARQ 197 plus erlotinib with erlotinib/placebo, in endothelial growth factor receptor inhibitor-naive patients with locally advanced/metastatic non-small cell lung cancer, demonstrate improvement in progression-free and overall survival with combined therapy. Results were especially pronounced for patients with non-squamous lung cancer histologies, and in particular molecularly defined subgroups including KRAS mutations. These and other data from ARQ 197 clinical trials in hepatocellular, germ-cell, pancreatic (in combination with gemcitabine), and colorectal (in combination with cetuximab and irinotecan) cancers further highlight the potential role of ARQ 197 in existing and emerging anticancer therapeutic regimens. PMID- 21632450 TI - Utility of mTOR inhibition in hematologic malignancies. AB - The mammalian target of rapamycin (mTOR) is an intracellular serine/threonine kinase that exists as a downstream component of numerous signaling pathways. The activation of mTOR results in the production of proteins involved in cell metabolism, growth, proliferation, and angiogenesis. Aberrant activation of mTOR signaling has been identified in a number of cancers, and targeted inhibition of mTOR has been successful in achieving tumor responses, prolonging progression free survival, and increasing overall survival in various oncologic patient populations. In particular, persistent activation of mTOR signaling has been identified in cell lines and patient samples with leukemias, Hodgkin's lymphoma (HL), non-Hodgkin's lymphoma (NHL), multiple myeloma (MM), and Waldenstrom's macroglobulinemia (WM). In vitro and preclinical studies using agents that inhibit mTOR signaling have demonstrated cytostatic and cytotoxic effects in these hematologic malignancies, suggesting that mTOR is a rational target for therapy in these disease states. In addition, the combination of mTOR inhibitors with traditional therapies may help to overcome the development of resistance and may improve response rates over those seen with established regimens through synergistic or additive effects. Inhibitors of mTOR signaling currently are being investigated in clinical trials of hematologic malignancies as single agents and as components of combination regimens. Thus far, promising results have been seen with the application of mTOR inhibitors as single agents in patients with relapsed or refractory leukemia, HL, NHL, MM, and WM. PMID- 21632451 TI - Outcomes after combined modality therapy for EGFR-mutant and wild-type locally advanced NSCLC. AB - BACKGROUND: Epidermal growth factor receptor (EGFR) mutations identify a unique biological subtype of non-small cell lung cancer (NSCLC). Treatment outcomes for EGFR-mutant locally advanced NSCLC patients have not been well described. METHODS: We retrospectively examined outcomes after combined modality therapy including thoracic radiation therapy (RT) in 123 patients with locally advanced NSCLC and known EGFR mutation status. Outcomes were compared using Kaplan-Meier analysis, the log-rank test, and multivariate Cox regression models. RESULTS: All 123 patients underwent thoracic RT; 25% had tumors with EGFR mutations and 94% had stage III disease. Overall, 81% received chemotherapy concurrent with RT and 55% underwent surgical resection. With a median follow-up of 27.5 months, the overall survival (OS) rate was significantly higher in patients with EGFR-mutant tumors than in those with wild-type EGFR tumors (2-year estimate: 92.6% versus 69.0%; p = .04). The 2-year relapse-free survival and distant recurrence rates did not differ significantly by genotype. The 2-year locoregional recurrence rate (LRR) was significantly lower in EGFR-mutant than in wild-type EGFR patients (17.8% versus 41.7%; p = .005). EGFR-mutant genotype was associated with a lower risk for LRR on multivariate analysis, but not OS, after adjusting for surgery and other potential confounders. CONCLUSION: We observed that EGFR-mutant patients with locally advanced NSCLC treated with RT had lower rates of LRR than wild-type EGFR patients, raising the hypothesis that EGFR mutations may confer sensitivity to RT and/or chemotherapy. The association between mutation status and OS after combined modality therapy was less robust. Our data may serve as a useful baseline estimate of outcomes by EGFR genotype for future prospective studies. PMID- 21632452 TI - When hope is all there is left. PMID- 21632453 TI - Initial staging impact of fluorodeoxyglucose positron emission tomography/computed tomography in locally advanced breast cancer. AB - PURPOSE: Fluorodeoxyglucose positron emission tomography/computed tomography (PET/CT) may reveal distant metastases more accurately than conventional imaging (CT, skeletal scintigraphy, chest radiography). We hypothesized that patients diagnosed with stage III noninflammatory breast cancer (non-IBC) and IBC by conventional imaging with PET/CT have a better prognosis than patients diagnosed without PET/CT. PATIENTS AND METHODS: We retrospectively identified 935 patients with stage III breast cancer in 2000-2009. We compared the relapse-free survival (RFS) and overall survival (OS) times of patients diagnosed by conventional imaging with those of patients diagnosed by conventional imaging plus PET/CT. Univariate and multivariate Cox proportional hazards regression models were used to assess associations between survival and PET/CT. RESULTS: RFS and OS times were not significantly different between patients imaged with PET/CT and those imaged without PET/CT. However, the RFS time in IBC patients was significantly different between patients imaged with PET/CT and those imaged without PET/CT on both univariate (hazard ratio [HR], 0.43; p = .014) and multivariate (HR, 0.33; p = .004) analysis. There was a trend for a longer OS duration in IBC patients imaged with PET/CT. CONCLUSION: Among IBC patients, adding PET/CT to staging based on conventional imaging might detect patients with metastases that were not detected by conventional imaging. The use of conventional imaging with PET/CT for staging in non-IBC patients is not justified on the basis of these retrospective data. The use of conventional imaging plus PET/CT in staging IBC needs to be studied prospectively to determine whether it will improve prognosis. PMID- 21632454 TI - A pilot phase II study of valproic acid for treatment of low-grade neuroendocrine carcinoma. AB - INTRODUCTION: Notch1 has been shown to be a tumor suppressor in neuroendocrine tumors (NETs). Previous in vitro studies in NET cell lines have also suggested that valproic acid (VPA), a histone deacetylase inhibitor, can induce Notch1 and that Notch1 activation correlates with a decrease in tumor markers for NETs. Thus, this study aimed to evaluate the role of VPA in treating NETs and to determine whether VPA induced the Notch signaling pathway signaling in vivo. PATIENTS AND METHODS: Eight patients with low-grade NETs (carcinoid and pancreatic) were treated with 500 mg of oral VPA twice a day with dosing adjusted to maintain a goal VPA level between 50 and 100 MUg/mL. All patients were followed for 12 months or until disease progression. RESULTS: Notch1 signaling was absent in all tumors prior to treatment and was upregulated with VPA. One patient had an unconfirmed partial response and was noted to have a 40-fold increase in Notch1 mRNA levels. Four patients had stable disease as best response. Tumor markers improved in 5 out of 7 patients. Overall, treatment with VPA was well tolerated. CONCLUSION: . VPA activates Notch1 signaling in vivo and may have a role in treating low-grade NETs. PMID- 21632455 TI - Tumor status at 12 weeks predicts survival in advanced colorectal cancer: findings from NCCTG N9741. AB - PURPOSE: We explored the prognostic value of actual tumor measurements (TM) versus World Health Organization (WHO) criteria as three-level (responder, stable, and progression) and two-level (responder and non-responder) variables at 12 and 24 weeks as predictors of survival in Intergroup Trial N9741, a phase III trial in metastatic colorectal cancer (CRC). METHODS: All patients with measurable disease (N = 1,188) were included. The percentage changes in TM from baseline to 12 and 24 weeks were calculated. The prognostic values of TM versus WHO criteria (as three- and two-level variables) at 12 and 24 weeks were compared, using Cox models for overall survival (OS) in a landmark analysis, adjusting for baseline tumor size, performance status, and treatment arm. RESULTS: Tumor status at 12 weeks by WHO criteria (three or two levels) or actual TM were all strongly associated with OS. Actual TM provided no meaningful additional benefit compared with the three-level WHO criteria. Tumor status at 24 weeks was also strongly associated with survival, but added no additional prognostic value compared with the 12-week assessment. At 12 weeks, actual TM improved prognostic characterization of patients with WHO status of response, but provided no additional value in patients with stable disease or progression. CONCLUSIONS: In N9741, the use of actual TM, or following tumor status beyond 12 weeks, did not improve survival prediction compared with a single three-level response assessment at 12 weeks, suggesting that 12-week tumor status could be an appropriate phase II trial endpoint in metastatic CRC. PMID- 21632456 TI - Part 1: background, methodology, and clinical adoption of pharmacogenetics. AB - Equivalent drug doses may lead to wide interpatient variability with regard to drug response, reflected by differences in drug activity and normal tissue toxicity. A major factor responsible for this variability is variation among patients in their genetic constitution. Genetic polymorphism may affect the activity of proteins encoded, which in turn may lead to changes in the pharmacokinetic and pharmacodynamic behavior of a drug, observed as differences in drug transport, drug metabolism, and pharmacodynamic drug effects. Recent insights into the functional effect of polymorphism in genes that are involved in the pharmacokinetics and pharmacodynamics of anticancer drugs have provided opportunities for patient-tailored therapy in oncology. Individualized pharmacotherapy based on genotype will help to increase treatment efficacy while reducing unnecessary toxicity, especially of drugs characterized by a narrow therapeutic window, such as anticancer drugs. We provide a series of four reviews aimed at implementing pharmacogenetic-based drug and dose prescription in the daily clinical setting for the practicing oncologist. This first part in the series describes the functional impact of genetic polymorphism and provides a general background to and insight into possible clinical consequences of pharmacogenetic variability. It also discusses different methodologies for clinical pharmacogenetic studies and provides a concise overview about the different laboratory technologies for genetic mutation analysis that are currently widely applied. Subsequently, pharmacogenetic association studies in anticancer drug transport, phase I and II drug metabolism, and pharmacodynamic drug effects are discussed in the rest of the series. Opportunities for patient tailored pharmacotherapy are highlighted. PMID- 21632457 TI - Cutaneous melanoma in situ: translational evidence from a large population-based study. AB - BACKGROUND: Cutaneous melanoma in situ (CMIS) is a nosologic entity surrounded by health concerns and unsolved debates. We aimed to shed some light on CMIS by means of a large population-based study. METHODS: Patients with histologic diagnosis of CMIS were identified from the Surveillance Epidemiology End Results (SEER) database. RESULTS: The records of 93,863 cases of CMIS were available for analysis. CMIS incidence has been steadily increasing over the past 3 decades at a rate higher than any other in situ or invasive tumor, including invasive skin melanoma (annual percentage change [APC]: 9.5% versus 3.6%, respectively). Despite its noninvasive nature, CMIS is treated with excision margins wider than 1 cm in more than one third of cases. CMIS is associated with an increased risk of invasive melanoma (standardized incidence ratio [SIR]: 8.08; 95% confidence interval [CI]: 7.66-8.57), with an estimated 3:5 invasive/in situ ratio; surprisingly, it is also associated with a reduced risk of gastrointestinal (SIR: 0.78, CI: 0.72-0.84) and lung (SIR: 0.65, CI: 0.59-0.71) cancers. Relative survival analysis shows that persons with CMIS have a life expectancy equal to that of the general population. CONCLUSIONS: CMIS is increasingly diagnosed and is often overtreated, although it does not affect the life expectancy of its carriers. Patients with CMIS have an increased risk of developing invasive melanoma (which warrants their enrollment in screening programs) but also a reduced risk of some epithelial cancers, which raises the intriguing hypothesis that genetic/environmental risk factors for some tumors may oppose the pathogenesis of others. PMID- 21632458 TI - Choosing the best second-line tyrosine kinase inhibitor in imatinib-resistant chronic myeloid leukemia patients harboring Bcr-Abl kinase domain mutations: how reliable is the IC50? AB - Development of drug resistance to imatinib mesylate in chronic myeloid leukemia (CML) patients is often accompanied by selection of point mutations in the kinase domain (KD) of the Bcr-Abl oncoprotein, where imatinib binds. Several second generation tyrosine kinase inhibitors (TKIs) have been designed rationally so as to enhance potency and retain the ability to bind mutated forms of Bcr-Abl. Since the preclinical phase of their development, most of these inhibitors have been tested in in vitro studies to assess their half maximal inhibitory concentration (IC50) for unmutated and mutated Bcr-Abl-that is, the drug concentration required to inhibit the cell proliferation or the phosphorylation processes driven by either the unmutated or the mutated forms of the kinase. A number of such studies have been published, and now that two inhibitors-dasatinib and nilotinib-are available for the treatment of imatinib-resistant cases, it is tempting for clinicians to reason on the IC50 values to guess, case by case, which one will work best in patients harboring specific Bcr-Abl KD mutations. Here, we discuss the pros and cons of using this approach in TKI selection. PMID- 21632460 TI - Trastuzumab. AB - Trastuzumab is standard of care in the treatment of human epidermal growth factor receptor (HER)-2+ early and advanced breast cancer. Recently, it has been approved for the treatment of HER-2+ advanced gastric cancer. Trastuzumab is an IgG1 humanized monoclonal antibody administered by intravenous infusion on a weekly or three weekly schedule. In all registered indications, trastuzumab is almost always given in combination with chemotherapy. In hormonal receptor positive breast cancer in postmenopausal women, trastuzumab can be combined with an aromatase inhibitor. Main toxicity is reduction in the left ventricular ejection fraction, which in a minority of patients can become symptomatic, but in many patients is at least partly reversible. Long-term safety needs to be further determined. PMID- 21632459 TI - Targeting angiogenesis in esophagogastric adenocarcinoma. AB - The possibility of targeting tumor angiogenesis was postulated almost 40 years ago. The vascular endothelial growth factor (VEGF) family and its receptors have since been characterized and extensively studied. VEGF overexpression is a common finding in solid tumors, including esophagogastric cancer, and frequently correlates with poor prognosis. Monoclonal antibodies, soluble receptors, and small-molecule tyrosine kinase inhibitors have been developed to inhibit tumor angiogenesis, and antiangiogenic therapy is now a component of standard treatment for advanced renal cell, hepatocellular, colorectal, breast, and non-small cell lung carcinomas. The small-molecule tyrosine kinase inhibitors sunitinib and sorafenib have been evaluated in phase II studies in esophagogastric cancer but appear to have only modest activity. Similarly, despite promising efficacy signals from phase II studies, the addition of the anti-VEGF-A monoclonal antibody bevacizumab to cisplatin plus capecitabine failed to result in a longer overall survival duration than with the chemotherapy doublet plus placebo. The response rate and progression-free survival interval were significantly greater with bevacizumab, confirming some efficacy in advanced gastric cancer, but with inadequate benefit to justify the high cost of treatment. Evaluation of bevacizumab in the neoadjuvant and perioperative settings continues, hypothesizing that a higher response rate will translate into longer survival in patients with operable disease. Despite extensive research, the discovery of a reliable predictive biomarker for antiangiogenic therapy continues to elude the scientific and oncology communities, and mechanisms of primary and acquired resistance are incompletely understood. We are therefore currently unable to personalize antiangiogenic therapy for established indications, or use molecular selection for clinical trials evaluating novel indications. PMID- 21632462 TI - Sodium butyrate inhibits the self-renewal capacity of endometrial tumor side population cells by inducing a DNA damage response. AB - We previously isolated side-population (SP) cells from a human endometrial cancer cell line, Hec1, and determined that Hec1-SP cells have cancer stem-like cell features. In this study, we isolated SP cells and non-SP (NSP) cells derived from a rat endometrial cell line expressing human [(12)Val] KRAS (RK12V cells) and determined the SP phenotype. RK12V-SP cells showed self-renewal capacity, the potential to develop into stromal cells, reduced expression levels of differentiation markers, long-term proliferating capacity in cultures, and enhanced tumorigenicity, indicating that RK12V-SP cells have cancer stem-like cell features. RK12V-SP cells also display higher resistance to conventional chemotherapeutic drugs. In contrast, treatment with a histone deacetylases (HDAC) inhibitor, sodium butyrate (NaB), reduced self-renewal capacity and completely suppressed colony formation of RK12V-SP cells in a soft agar. The levels of intracellular reactive oxygen species (ROS) and the number of gammaH2AX foci were increased by NaB treatment of both RK12V-SP cells and RK12V-NSP cells. The expression levels of gammaH2AX, p21, p27, and phospho-p38 mitogen-activated protein kinase were enhanced in RK12V-SP cells compared with RK12V-NSP cells. These results imply that treatment with NaB induced production of intracellular ROS and DNA damage in both RK12V-SP and RK12V-NSP cells. Following NaB treatment, DNA damage response signals were enhanced more in RK12V-SP cells than in RK12V NSP cells. This is the first article on an inhibitory effect of NaB on proliferation of endometrial cancer stem-like cells. HDAC inhibitors may represent an attractive antitumor therapy based upon their inhibitory effects on cancer stem-like cells. PMID- 21632463 TI - In vivo activity of combined PI3K/mTOR and MEK inhibition in a Kras(G12D);Pten deletion mouse model of ovarian cancer. AB - The phosphatidylinositol 3-kinase (PI3K)/Akt pathway is commonly dysregulated in human cancer, making it an attractive target for novel anticancer therapeutics. We have used a mouse model of ovarian cancer generated by Kras(G12D) activation and Pten deletion in the ovarian surface epithelium for the preclinical assessment of a novel PI3K/mTOR inhibitor PF-04691502. To enable higher throughput studies, we developed an orthotopic primary transplant model from these mice and evaluated therapeutic response to PF-04691502 using small-animal ultrasound and FDG-PET imaging. PF-04691502 inhibited tumor growth at 7 days by 72% +/- 9. FDG-PET imaging revealed that PF-04691502 reduced glucose metabolism dramatically, suggesting FDG-PET may be exploited as an imaging biomarker of target inhibition by PF-04691502. Tissue biomarkers of PI3K/mTOR pathway activity, p-AKT (S473), and p-RPS6 (S240/244), were also dramatically inhibited following PF-04691502 treatment. However, as a single agent, PF-04691502 did not induce tumor regression and the long-term efficacy was limited, with tumor proliferation continuing in the presence of drug treatment. We hypothesized that tumor progression was because of concomitant activation of the mitogen-activated protein kinase pathway downstream of Kras(G12D) expression promoting cell survival and that the therapeutic effect of PF-04691502 would be enhanced by combinatory inhibition of MEK using PD-0325901. This combination induced striking tumor regression, apoptosis associated with upregulation of Bim and downregulation of Mcl-1, and greatly improved duration of survival. These data suggest that contemporaneous MEK inhibition enhances the cytotoxicity associated with abrogation of PI3K/mTOR signaling, converting tumor growth inhibition to tumor regression in a mouse model of ovarian cancer driven by PTEN loss and mutant K-Ras. PMID- 21632461 TI - Part 2: pharmacogenetic variability in drug transport and phase I anticancer drug metabolism. AB - Equivalent drug doses in anticancer chemotherapy may lead to wide interpatient variability in drug response reflected by differences in treatment response or in severity of adverse drug reactions. Differences in the pharmacokinetic (PK) and pharmacodynamic (PD) behavior of a drug contribute to variation in treatment outcome among patients. An important factor responsible for this variability is genetic polymorphism in genes that are involved in PK/PD processes, including drug transporters, phase I and II metabolizing enzymes, and drug targets, and other genes that interfere with drug response. In order to achieve personalized pharmacotherapy, drug dosing and treatment selection based on genotype might help to increase treatment efficacy while reducing unnecessary toxicity. We present a series of four reviews about pharmacogenetic variability in anticancer drug treatment. This is the second review in the series and is focused on genetic variability in genes encoding drug transporters (ABCB1 and ABCG2) and phase I drug-metabolizing enzymes (CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP3A4, CYP3A5, DPYD, CDA and BLMH) and their associations with anticancer drug treatment outcome. Based on the literature reviewed, opportunities for patient-tailored anticancer therapy are presented. PMID- 21632464 TI - Phenotypic reversion of invasive neurofibromin-deficient schwannoma by FTS: Ras inhibition reduces BMP4/Erk/Smad signaling. AB - Neurofibromin-deficient (Nf1(-/-)) malignant peripheral nerve sheath tumors (MPNST) are highly invasive, refractory to chemotherapy, and characterized by overactivated Ras. Ras activates mitogenic pathways and regulates morphogenic programs--such as those induced by bone morphogenetic proteins (BMP) and TGF beta. The role of such a cross-talk in determining the phenotype and transformation potential of MPNSTs is unknown. Here, we used MPNST cell lines and selective Ras inhibition with S-trans,trans-farnesylthiosalicylic-acid (FTS; salirasib) in conjunction with specific inhibitors of TGF-beta and BMP signaling. FTS perturbed signaling of BMP4 and TGF-beta1 to Smad-dependent and Erk-dependent pathways. Furthermore, FTS inhibited motility and spreading, reduced the gelatinase secretion, eliminated the expression and activation of regulators of cell-matrix interaction, and altered gene expression. These phenomena are indicative of a phenotypic reversion of NF1-deficient cells by FTS. Inhibition of BMP4 and TGF-beta by noggin and SB-431542, respectively, mimicked the FTS mediated effects on adhesion, spreading, and cell morphology. This strongly suggests that a cross-talk among TGF-beta superfamily ligands and Ras plays a significant role in the transformation of NF1(-/-) MPNSTs. Our results support the therapeutic potential of FTS, in conjuncture with BMP and TGF-beta pathway inhibitors, toward the inhibition of mitogenic and morphogenic signaling pathways and the alleviation of NF1 symptoms. PMID- 21632465 TI - The Family-centered Action Model of Intervention Layout and Implementation (FAMILI): the example of childhood obesity. AB - Parents play a fundamental role in shaping children's development, including their dietary and physical activity behaviors. Yet family-centered interventions are rarely used in obesity prevention research. Less than half of childhood obesity prevention programs include parents, and those that do include parents or a family component seldom focus on sustainable change at the level of the family. The general absence of a family-centered approach may be explained by persistent challenges in engaging parents and families and the absence of an intervention framework explicitly designed to foster family-centered programs. The Family centered Action Model of Intervention Layout and Implementation, or FAMILI, was developed to address these needs. FAMILI draws on theories of family development to frame research and intervention design, uses a mixed-methods approach to conduct ecologically valid research, and positions family members as active participants in the development, implementation, and evaluation of family centered obesity prevention programs. FAMILI is intended to facilitate the development of culturally responsive and sustainable prevention programs with the potential to improve outcomes. Although childhood obesity was used to illustrate the application of FAMILI, this model can be used to address a range of child health problems. PMID- 21632466 TI - Fasting insulin reflects heterogeneous physiological processes: role of insulin clearance. AB - Several processes contribute to variation in fasting insulin concentration, including fasting glucose, insulin resistance, insulin secretion, and insulin clearance. Our goal was to determine the relative contribution of each of these insulin-related traits, plus anthropometric parameters, to fasting insulin among 470 Mexican Americans. The euglycemic hyperinsulinemic clamp yielded insulin sensitivity (M value) and metabolic clearance rate of insulin (MCRI). Acute insulin secretion was estimated by the insulinogenic index (IGI30) from the oral glucose tolerance test. Regression (univariate) and generalized estimating equations (multivariate) were used to describe the relationship of insulin related traits to fasting insulin. Univarate analyses were used to select which traits to include in the multivariate model. In multivariate analysis, MCRI, M, BMI, waist circumference, and fasting glucose were independently associated with fasting insulin. Decreasing M and MCRI were associated with increasing fasting insulin, whereas increasing BMI, waist circumference, and fasting glucose were associated with increasing fasting insulin. Standardized coefficients allowed determination of the relative strength of each trait's association with fasting insulin in the entire cohort (strongest to weakest): MCRI (-0.35, P < 0.0001), M (-0.24, P < 0.0001), BMI (0.20, P = 0.0011), waist circumference (0.16, P = 0.021), and fasting glucose (0.11, P = 0.014). Fasting insulin is a complex phenotype influenced by several independent processes, each of which might have its own environmental and genetic determinants. One of the most associated traits was insulin clearance, which has implications for studies that have used fasting insulin as a surrogate for insulin resistance. PMID- 21632467 TI - Cancer cells exploit the Eph-ephrin system to promote invasion and metastasis: tales of unwitting partners. AB - The Eph subfamily of receptor tyrosine kinases and their membrane-anchored ephrin ligands mediate cell-cell contact signaling and are versatile regulators of cell migration and tissue patterning, which are often exploited by cancer cells during tumor progression. New evidence shows that prostate cancer cells use EphA2 and EphA4 receptors and ephrin-As to mediate homotypic contact inhibition of locomotion while co-opting ephrin-B2 on stromal cells through EphB3 and EphB4 receptors to propel migration. These processes could enhance cancer cell scattering from the primary tumor mass and promote unimpeded migration and invasion through the stromal space. The results provide another example in which Eph receptors are converted into pro-oncogenic proteins, contrary to their often described tumor suppressor roles in normal tissues. PMID- 21632468 TI - Reduction of complex signaling networks to a representative kernel. AB - The network of biomolecular interactions that occurs within cells is large and complex. When such a network is analyzed, it can be helpful to reduce the complexity of the network to a "kernel" that maintains the essential regulatory functions for the output under consideration. We developed an algorithm to identify such a kernel and showed that the resultant kernel preserves the network dynamics. Using an integrated network of all of the human signaling pathways retrieved from the KEGG (Kyoto Encyclopedia of Genes and Genomes) database, we identified this network's kernel and compared the properties of the kernel to those of the original network. We found that the percentage of essential genes to the genes encoding nodes outside of the kernel was about 10%, whereas ~32% of the genes encoding nodes within the kernel were essential. In addition, we found that 95% of the kernel nodes corresponded to Mendelian disease genes and that 93% of synthetic lethal pairs associated with the network were contained in the kernel. Genes corresponding to nodes in the kernel had low evolutionary rates, were ubiquitously expressed in various tissues, and were well conserved between species. Furthermore, kernel genes included many drug targets, suggesting that other kernel nodes may be potential drug targets. Owing to the simplification of the entire network, the efficient modeling of a large-scale signaling network and an understanding of the core structure within a complex framework become possible. PMID- 21632469 TI - Integration of activating and inhibitory receptor signaling by regulated phosphorylation of Vav1 in immune cells. AB - Natural killer (NK) cells are effector cells of the immune system whose activation is carefully regulated by the interplay of signals from activating and inhibitory receptors. Signals from activating receptors induce phosphorylation of the guanine nucleotide exchange factor Vav1, whereas those from inhibitory receptors lead to the dephosphorylation of Vav1 by the Src homology 2 domain containing protein tyrosine phosphatase 1 (SHP-1). Here, we used mathematical modeling and experiments with NK cells to gain insight into this integration of positive and negative signals at a molecular level. Our data showed a switch-like regulation of Vav1 phosphorylation, the extent of which correlated with the cytotoxic activity of NK cells. Comparison of our experimental results with the predictions that we derived from an ensemble of 72 mathematical models showed that a physical association between Src family kinases and activating receptors on NK cells was essential to generate the cytotoxic response. Our data support a central role for Vav1 in determining the cytotoxic activity of NK cells and provide insight into the molecular mechanism of the integration of positive and negative signals during lymphocyte activation. PMID- 21632470 TI - Diagnosis and management of sports-related concussion: a 15-year-old athlete with a concussion. AB - Concussion in youth athletes is a growing problem worldwide. During the past decade, significant progress has been made in standardization of the assessment of young athletes, and a growing appreciation of metabolic vulnerability, activity, and cognitive challenges has led to guidelines and suggestions for rest from the field as well as cognitive rest from school. Outcome data have begun to establish groups linked to symptom class, genetics, and sex who are at risk of worse outcomes from concussions. Decisions regarding return to activity are now based on at-rest symptoms, graded increases in activity, and neuropsychological testing. Using the case of Ms X, a 15-year-old otherwise healthy high school student who fell while skiing, evaluation, prognosis, and management of concussion are discussed. PMID- 21632471 TI - Elimination of severe albuminuria in aging hypertensive rats by exchange of 2 chromosomes in double-consomic rats. AB - The inherited nephron deficit and progressive albuminuria development observed in hypertensive Munich Wistar Fromter (MWF) rats are influenced by quantitative trait loci on rat chromosome (RNO) 6 and RNO8. Previous studies in young MWF rats suggested that the nephron deficit represents a cause for glomerular hypertrophy preceding onset of albuminuria at 8 weeks and demonstrated a simultaneous induction of the podocyte stress marker desmin and podoplanin loss in podocytes. Here we investigated the separate genetic influence of RNO6 and RNO8 on early glomerular changes and subsequent albuminuria in single-consomic MWF rats in which RNO6 (MWF-6(SHR)) and RNO8 (MWF-8(SHR)) were replaced by the respective spontaneously hypertensive rat (SHR) chromosome. Furthermore, we tested the role of synergistic effects between both chromosomes in a double-consomic MWF 6(SHR)8(SHR) strain. Increased glomerular, extramesangial desmin expressions at 6 and albuminuria at 8 weeks were significantly reduced in single- and double consomics (P<0.05 versus MWF, respectively). MWF-6(SHR)8(SHR) rats demonstrated the lowest desmin expression and glomerular volume (P<0.05 versus MWF, MWF 6(SHR), and MWF-8(SHR), respectively), indicating synergistic effects between RNO6 and RNO8. A significant and similar loss of podoplanin was only seen in MWF and MWF-6(SHR) rats but not in MWF-8(SHR) and MWF-6(SHR)8(SHR) rats (P<0.02, respectively); this refutes a mandatory coupling of desmin induction and podoplanin loss in podocytes preceding albuminuria and reveals a genetic link between RNO8 and loss of podoplanin protein. Long-term follow up in MWF 6(SHR)8(SHR) rats demonstrates the relevance of the absence of glomerular changes in young animals, because double-consomics demonstrate a complete suppression of progressive albuminuria and kidney damage compared with MWF rats despite similar blood pressures. PMID- 21632472 TI - Effect of a reduction in uric acid on renal outcomes during losartan treatment: a post hoc analysis of the reduction of endpoints in non-insulin-dependent diabetes mellitus with the Angiotensin II Antagonist Losartan Trial. AB - Emerging data show that increased serum uric acid (SUA) concentration is an independent risk factor for end-stage renal disease. Treatment with the antihypertensive drug losartan lowers SUA. Whether reductions in SUA during losartan therapy are associated with renoprotection is unclear. We therefore tested this hypothesis. In a post hoc analysis of 1342 patients with type 2 diabetes mellitus and nephropathy participating in the Reduction of Endpoints in Non-Insulin-Dependent Diabetes Mellitus With the Angiotensin II Antagonist Losartan Trial, we determined the relationship between month 6 change in SUA and renal endpoints, defined as a doubling of serum creatinine or end-stage renal disease. Baseline SUA was 6.7 mg/dL in placebo and losartan-treated subjects. During the first 6 months, losartan lowered SUA by -0.16 mg/dL (95% CI: -0.30 to 0.01; P=0.031) as compared with placebo. The risk of renal events was decreased by 6% (95% CI: 10% to 3%) per 0.5-mg/dL decrement in SUA during the first 6 months. This effect was independent of other risk markers, including estimate glomerular filtration rate and albuminuria. Adjustment of the overall treatment effects for SUA attenuated losartan's renoprotective effect from 22% (95% CI: 6% to 35%) to 17% (95% CI: 1% to 31%), suggesting that approximately one fifth of losartan's renoprotective effect could be attributed to its effect on SUA. Losartan lowers SUA levels compared with placebo treatment in patients with type 2 diabetes mellitus and nephropathy. The degree of reduction in SUA is subsequently associated with the degree in long-term renal risk reduction and explains part of losartan's renoprotective effect. These findings support the view that SUA may be a modifiable risk factor for renal disease. PMID- 21632473 TI - Diurnal variation in the mechanical and neural components of the baroreflex. AB - Diminished baroreflex sensitivity in the morning negatively influences morning coronary blood flow and blood pressure control in hypertensive patients. Our aim was to determine the contribution of the mechanical and neural components of the cardiac baroreflex to diurnal variation in blood pressure control. In 12 healthy participants, we used the modified Oxford method to quantify baroreflex sensitivity for rising (G(up)) and falling (G(down)) pressures in the morning (7:00 am) and afternoon (4:00 pm). Beat-to-beat blood pressure, R-R intervals, and carotid artery diameter measurements were recorded. Integrated sensitivity was determined by plotting R-R intervals against systolic blood pressure. The mechanical component was carotid artery diameter plotted against systolic blood pressure, and the neural component was R-R intervals plotted against carotid artery diameter. Linear mixed models were used to compare the integrated, mechanical, and neural sensitivities between morning and afternoon. We found significant diurnal variation in integrated sensitivity, with an attenuated response in the morning (G(up)=13.0+/-0.6; G(down)=6.3+/-0.4 ms/mm Hg) when compared with the afternoon (G(up)=15.1+/-0.6; G(down)=12.6+/-0.4 ms/mm Hg). For rising pressures, the diminished integrated sensitivity in the morning was caused by a reduction in mechanical sensitivity, whereas for falling pressures it was caused by a reduction in neural sensitivity. Our findings explicate the mechanisms underlying diurnal variation in baroreflex function. Pharmacological and lifestyle interventions targeted specifically at the diminished component of the cardiac baroreflex in the morning may lead to better management of hypertension. PMID- 21632474 TI - The cover. Blue courtyard. PMID- 21632475 TI - A piece of my mind. A house built out of madness. PMID- 21632476 TI - The National Library of Medicine: 175 years of advancing biomedical knowledge. PMID- 21632477 TI - Study: Up to 1 in 10 infants given herbal supplements, teas by their mother. PMID- 21632478 TI - Antihypertensive treatment of patients with cardiovascular disease but without hypertension. PMID- 21632479 TI - Antihypertensive treatment of patients with cardiovascular disease but without hypertension. PMID- 21632480 TI - Stroke after coronary artery bypass graft surgery. PMID- 21632481 TI - Effect of 1 week of sleep restriction on testosterone levels in young healthy men. PMID- 21632483 TI - Association of operative time of day with outcomes after thoracic organ transplant. AB - CONTEXT: Recent emphasis on systems-based approaches to patient safety has led to several studies demonstrating worse outcomes associated with surgery at night. OBJECTIVE: To evaluate whether operative time of day was associated with thoracic organ transplant outcomes, hypothesizing that it would not be associated with increased morbidity or mortality. DESIGN, SETTING, AND PARTICIPANTS: We conducted a retrospective cohort study of adult heart and lung transplant recipients in the United Network for Organ Sharing database from January 2000 through June 2010. Primary stratification was by operative time of day (night, 7 PM-7 AM; day, 7 AM 7 PM). MAIN OUTCOME MEASURES: Primary end points were short-term survival, assessed by the Kaplan-Meier method at 30, 90, and 365 days. Secondary end points encompassed common postoperative complications. Risk-adjusted multivariable Cox proportional hazards regression examined mortality. RESULTS: A total of 27,118 patients were included in the study population. Of the 16,573 who underwent a heart transplant, 8346 (50.36%) did so during the day and 8227 (49.64%) during the night. Of the 10,545 who underwent a lung transplant, 5179 (49.11%) did so during the day and 5366 (50.89%) during the night. During a median follow-up of 32.2 months (interquartile range, 11.2-61.1 months), 8061 patients (28.99%) died. Survival was similar for organ transplants performed during the day and night. Survival rates at 30 days for heart transplants during the day were 95.0% vs 95.2% during the night (hazard ratio [HR], 1.05; 95% confidence interval, 0.83 1.32; P = .67) and for lung transplants during the day were 96.0% vs 95.5% during the night (HR, 1.22; 95% CI, 0.97-1.55; P = .09). At 90 days, survival rates for heart transplants were 92.6% during the day vs 92.7% during the night (HR, 1.05; 95% CI, 0.88-1.26; P = .59) and for lung transplants during the day were 92.7% vs 91.7% during the night (HR, 1.23; 95% CI, 1.04-1.47; P = .02). At 1 year, survival rates for heart transplants during the day were 88.0% vs 87.7% during the night (HR, 1.05; 95% CI, 0.91-1.21; P = .47) and for lung transplants during the day were 83.8% vs 82.6% during the night (HR, 1.08; 95% CI, 0.96-1.22; P = .19). Among lung transplant recipients, there was a slightly higher rate of airway dehiscence associated with nighttime transplants (57 of 5022 [1.1%] vs 87 of 5224 [1.7%], P = .02). CONCLUSION: Among patients who underwent thoracic organ transplants, there was no significant association between operative time of day and survival up to 1 year after organ transplant. PMID- 21632484 TI - Comparison of effect sizes associated with biomarkers reported in highly cited individual articles and in subsequent meta-analyses. AB - CONTEXT: Many biomarkers are proposed in highly cited studies as determinants of disease risk, prognosis, or response to treatment, but few eventually transform clinical practice. OBJECTIVE: To examine whether the magnitude of the effect sizes of biomarkers proposed in highly cited studies is accurate or overestimated. DATA SOURCES: We searched ISI Web of Science and MEDLINE until December 2010. STUDY SELECTION: We included biomarker studies that had a relative risk presented in their abstract. Eligible articles were those that had received more than 400 citations in the ISI Web of Science and that had been published in any of 24 highly cited biomedical journals. We also searched MEDLINE for subsequent meta-analyses on the same associations (same biomarker and same outcome). DATA EXTRACTION: In the highly cited studies, data extraction was focused on the disease/outcome, biomarker under study, and first reported relative risk in the abstract. From each meta-analysis, we extracted the overall relative risk and the relative risk in the largest study. Data extraction was performed independently by 2 investigators. RESULTS: We evaluated 35 highly cited associations. For 30 of the 35 (86%), the highly cited studies had a stronger effect estimate than the largest study; for 3 the largest study was also the highly cited study; and only twice was the effect size estimate stronger in the largest than in the highly cited study. For 29 of the 35 (83%) highly cited studies, the corresponding meta-analysis found a smaller effect estimate. Only 15 of the associations were nominally statistically significant based on the largest studies, and of those only 7 had a relative risk point estimate greater than 1.37. CONCLUSION: Highly cited biomarker studies often report larger effect estimates for postulated associations than are reported in subsequent meta analyses evaluating the same associations. PMID- 21632482 TI - Association of BMD and FRAX score with risk of fracture in older adults with type 2 diabetes. AB - CONTEXT: Type 2 diabetes mellitus (DM) is associated with higher bone mineral density (BMD) and paradoxically with increased fracture risk. It is not known if low BMD, central to fracture prediction in older adults, identifies fracture risk in patients with DM. OBJECTIVE: To determine if femoral neck BMD T score and the World Health Organization Fracture Risk Algorithm (FRAX) score are associated with hip and nonspine fracture risk in older adults with type 2 DM. DESIGN, SETTING, AND PARTICIPANTS: Data from 3 prospective observational studies with adjudicated fracture outcomes (Study of Osteoporotic Fractures [December 1998 July 2008]; Osteoporotic Fractures in Men Study [March 2000-March 2009]; and Health, Aging, and Body Composition study [April 1997-June 2007]) were analyzed in older community-dwelling adults (9449 women and 7436 men) in the United States. MAIN OUTCOME MEASURE: Self-reported incident fractures, which were verified by radiology reports. RESULTS: Of 770 women with DM, 84 experienced a hip fracture and 262 a nonspine fracture during a mean (SD) follow-up of 12.6 (5.3) years. Of 1199 men with DM, 32 experienced a hip fracture and 133 a nonspine fracture during a mean (SD) follow-up of 7.5 (2.0) years. Age-adjusted hazard ratios (HRs) for 1-unit decrease in femoral neck BMD T score in women with DM were 1.88 (95% confidence interval [CI], 1.43-2.48) for hip fracture and 1.52 (95% CI, 1.31-1.75) for nonspine fracture, and in men with DM were 5.71 (95% CI, 3.42-9.53) for hip fracture and 2.17 (95% CI, 1.75-2.69) for nonspine fracture. The FRAX score was also associated with fracture risk in participants with DM (HRs for 1-unit increase in FRAX hip fracture score, 1.05; 95% CI, 1.03-1.07, for women with DM and 1.16; 95% CI, 1.07-1.27, for men with DM; HRs for 1-unit increase in FRAX osteoporotic fracture score, 1.04; 95% CI, 1.02-1.05, for women with DM and 1.09; 95% CI, 1.04-1.14, for men with DM). However, for a given T score and age or for a given FRAX score, participants with DM had a higher fracture risk than those without DM. For a similar fracture risk, participants with DM had a higher T score than participants without DM. For hip fracture, the estimated mean difference in T score for women was 0.59 (95% CI, 0.31-0.87) and for men was 0.38 (95% CI, 0.09-0.66). CONCLUSIONS: Among older adults with type 2 DM, femoral neck BMD T score and FRAX score were associated with hip and nonspine fracture risk; however, in these patients compared with participants without DM, the fracture risk was higher for a given T score and age or for a given FRAX score. PMID- 21632485 TI - A new frontier in patient safety. PMID- 21632486 TI - The physics of health care improvement. PMID- 21632487 TI - Comparative effectiveness--looking under the lamppost. PMID- 21632488 TI - The thin line between hope and hype in biomarker research. PMID- 21632489 TI - JAMA patient page. BRCA genes and breast cancer. PMID- 21632490 TI - Calcineurin splicing variant calcineurin Abeta1 improves cardiac function after myocardial infarction without inducing hypertrophy. AB - BACKGROUND: Calcineurin is a calcium-regulated phosphatase that plays a major role in cardiac hypertrophy. We previously described that alternative splicing of the calcineurin Abeta (CnAbeta) gene generates the CnAbeta1 isoform, with a unique C-terminal region that is different from the autoinhibitory domain present in all other CnA isoforms. In skeletal muscle, CnAbeta1 is necessary for myoblast proliferation and stimulates regeneration, reducing fibrosis and accelerating the resolution of inflammation. Its role in the heart is currently unknown. METHODS AND RESULTS: We generated transgenic mice overexpressing CnAbeta1 in postnatal cardiomyocytes under the control of the alpha-myosin heavy chain promoter. In contrast to previous studies using an artificially truncated calcineurin, CnAbeta1 overexpression did not induce cardiac hypertrophy. Moreover, transgenic mice showed improved cardiac function and reduced scar formation after myocardial infarction, with reduced neutrophil and macrophage infiltration and decreased expression of proinflammatory cytokines. Immunoprecipitation and Western blot analysis showed interaction of CnAbeta1 with the mTOR complex 2 and activation of the Akt/SGK cardioprotective pathway in a PI3K-independent manner. In addition, gene expression profiling revealed that CnAbeta1 activated the transcription factor ATF4 downstream of the Akt/mTOR pathway to promote the amino acid biosynthesis program, to reduce protein catabolism, and to induce the antifibrotic and antiinflammatory factor growth differentiation factor 15, which protects the heart through Akt activation. CONCLUSIONS: Calcineurin Abeta1 shows a unique mode of action that improves cardiac function after myocardial infarction, activating different cardioprotective pathways without inducing maladaptive hypertrophy. These features make CnAbeta1 an attractive candidate for the development of future therapeutic approaches. PMID- 21632491 TI - Sensitive cardiac troponin T assay and the risk of incident cardiovascular disease in women with and without diabetes mellitus: the Women's Health Study. AB - BACKGROUND: Very low levels of cardiac troponin T are associated with an increased risk of cardiovascular death in patients with stable chronic coronary disease. Whether high-sensitivity cardiac troponin T levels are associated with adverse cardiovascular outcomes in individuals without cardiovascular disease (CVD) has not been well studied. METHODS AND RESULTS: Using 2 complementary study designs, we evaluated the relationship between baseline cardiac troponin and incident CVD events among diabetic and nondiabetic participants in the Women's Health Study (median follow-up, 12.3 years). All diabetic women with blood specimens were included in a cohort study (n=512 diabetic women, n=65 events), and nondiabetic women were sampled for inclusion in a case-cohort analysis (n=564 comprising the subcohort, n=479 events). High-sensitivity cardiac troponin T was detectable (>= 0.003 MUg/L) in 45.5% of diabetic women and 30.3% of nondiabetic women (P<0.0001). In models adjusted for traditional risk factors and hemoglobin A(1c), detectable high-sensitivity cardiac troponin T was associated with subsequent CVD (myocardial infarction, stroke, cardiovascular death) in diabetic women (adjusted hazard ratio, 1.79; 95% confidence interval, 1.04 to 3.07, P=0.036) but not nondiabetic women (adjusted hazard ratio, 1.13; 95% confidence interval, 0.82 to 1.55; P=0.46). Further adjustment for amino-terminal pro-B-type natriuretic peptide and estimated renal function did not substantially alter this relationship among diabetic women (hazard ratio, 1.76; 95% confidence interval, 1.00 to 3.08; P=0.0499), which appeared to be driven by a 3-fold increase in CVD death that was not observed in nondiabetic women. CONCLUSIONS: Very low but detectable levels of cardiac troponin T are associated with total CVD and CVD death in women with diabetes mellitus. Among healthy nondiabetic women, detectable compared with undetectable troponin was not associated with CVD events. PMID- 21632492 TI - Differences in admitting hospital characteristics for black and white Medicare beneficiaries with acute myocardial infarction. AB - BACKGROUND: Racial disparities in acute myocardial infarction treatment may be due to differences in admitting hospitals. Little is known about factors associated with hospital selection for black and white acute myocardial infarction patients. METHODS AND RESULTS: We identified black and white Medicare beneficiaries with acute myocardial infarction in 63 hospital referral regions with at least 50 black admissions during 2005 (n=65,633). We calculated distance from patient home to hospital referral region hospitals using ZIP code centroids. We assessed hospital quality using a composite score made up of hospital risk adjusted 30-day mortality and acute myocardial infarction performance measures. Hospitals with a score in the top 20% were categorized as high quality, and those in the lowest 20% as low quality. We used conditional multinomial logit models to examine differences in hospital selection for blacks and whites. On average, blacks lived closer to revascularization hospitals (mean, 3.8 versus 6.8 miles; P<0.001) and to high-quality hospitals (mean, 5.6 versus 9.7 miles; P<0.001). After distance was accounted for, blacks were relatively less likely (P<0.001) to be admitted to revascularization hospitals (risk ratio [RR], 0.87; 95% confidence interval [CI], 0.80 to 0.95) and to high-quality hospitals (RR, 0.88; 95% CI, 0.801 to 0.95) but more likely (P<0.001) to be admitted to low-quality hospitals (RR, 1.17; 95% CI, 1.05 to 1.29). In analyses matched by home ZIP code, differences in admissions to revascularization (RR, 0.92; 95% CI, 0.80 to 1.05), high-quality (RR, 0.94; 95% CI, 0.81 to 1.07), and low-quality (RR, 1.15; 95% CI, 0.94 to 1.35) hospitals were not significant. CONCLUSIONS: Differences in admissions to revascularization and high-quality hospitals may contribute to disparities in acute myocardial infarction care. These differences may be due in part to residential ZIP code characteristics. PMID- 21632493 TI - Early repolarization: electrocardiographic phenotypes associated with favorable long-term outcome. AB - BACKGROUND: Early repolarization (ER) in inferior/lateral leads of standard ECGs increases the risk of arrhythmic death. We tested the hypothesis that variations in the ST-segment characteristics after the ER waveforms may have prognostic importance. METHODS AND RESULTS: ST segments after ER were classified as horizontal/descending or rapidly ascending/upsloping on the basis of observations from 2 independent samples of young healthy athletes from Finland (n=62) and the United States (n=503), where ascending type was the dominant and common form of ER. Early repolarization was present in 27/62 (44%) of the Finnish athletes and 151/503 (30%) of the US athletes, and all but 1 of the Finnish (96%) and 91/107 (85%) of US athletes had an ascending/upsloping ST variant after ER. Subsequently, ECGs from a general population of 10 864 middle-aged subjects were analyzed to assess the prognostic modulation of ER-associated risk by ST-segment variations. Subjects with ER >=0.1 mV and horizontal/descending ST variant (n=412) had an increased hazard ratio of arrhythmic death (relative risk 1.43; 95% confidence interval 1.05 to 1.94). When modeled for higher amplitude ER (>0.2 mV) in inferior leads and horizontal/descending ST-segment variant, the hazard ratio of arrhythmic death increased to 3.14 (95% confidence interval 1.56 to 6.30). However, in subjects with ascending ST variant, the relative risk for arrhythmic death was not increased (0.89; 95% confidence interval 0.52 to 1.55). CONCLUSIONS: ST-segment morphology variants associated with ER separates subjects with and without an increased risk of arrhythmic death in middle-aged subjects. Rapidly ascending ST segments after the J-point, the dominant ST pattern in healthy athletes, seems to be a benign variant of ER. PMID- 21632494 TI - Birth weight predicts risk of cardiovascular disease within dizygotic but not monozygotic twin pairs: a large population-based co-twin-control study. AB - BACKGROUND: The widely reported inverse association between birth weight and risk of cardiovascular disease (CVD) has sparked theories about early life determinants of adult disease. Within-twin-pair analysis provides a unique opportunity to investigate whether factors shared within twin pairs influence the association. METHODS AND RESULTS: In a population-based cohort of like-sexed twins with known zygosity born in Sweden from 1926 to 1958, disease-discordant twin pairs were identified through linkage to the National Inpatient and Cause of Death registers between 1973 and 2006. Co-twin-control analyses were performed on twins discordant for cardiovascular disease (n=3884), coronary heart disease (n=2668), and stroke (n=1372). Overall, inverse associations between birth weight and risk of cardiovascular diseases were seen within dizygotic but not monozygotic twin pairs. In dizygotic twins, the odds ratios for a 1-kg within pair increase in birth weight were 0.74 (95% confidence interval, 0.56 to 0.98) for coronary heart disease and 0.57 (95% confidence interval, 0.37 to 0.88) for stroke. Conversely, no statistically significant associations were found within monozygotic twins (for coronary heart disease: odds ratio, 1.10; 95% confidence interval, 0.73 to 1.68; for stroke: odds ratio, 0.92; 95% confidence interval, 0.48 to 1.80). CONCLUSIONS: We found an association between birth weight and risk of cardiovascular disease within disease-discordant dizygotic but not monozygotic twin pairs. This indicates that the association between birth weight and cardiovascular disease could be a result of common causes, and that factors that vary within dizygotic but not monozygotic twin pairs may help identify them. PMID- 21632495 TI - Risk of recurrent cardiac events after onset of menopause in women with congenital long-QT syndrome types 1 and 2. AB - BACKGROUND: Women with congenital long-QT syndrome experience an increased risk for cardiac events after the onset of adolescence that is more pronounced among carriers of the LQT2 genotype. We hypothesized that the hormonal changes associated with menopause may affect clinical risk in this population. METHODS AND RESULTS: We used a repeated-events analysis to evaluate the risk for recurrent syncope during the menopause transition and postmenopausal periods (5 years before and after the age at onset of menopause, respectively) among 282 LQT1 (n=151) and LQT2 (n=131) women enrolled in the Long-QT Syndrome Registry. Multivariate analysis showed that the risk for recurrent syncope (n=150) among LQT2 women was significantly increased during both menopause transition (hazard ratio, 3.38; P=0.005) and the postmenopausal period (hazard ratio, 8.10; P<0.001) compared with the reproductive period. The risk increase was evident among women who did or did not receive estrogen therapy. In contrast, among LQT1 women, the onset of menopause was associated with a reduction in the risk for recurrent syncope (hazard ratio, 0.19; P=0.05; P=0.02 for genotype-by-menopause interaction). Only 22 women (8%) experienced aborted cardiac arrest or sudden cardiac death during follow-up. The frequency of aborted cardiac arrest/sudden cardiac death showed a similar genotype-specific association with the onset of menopause. CONCLUSIONS: The onset of menopause is associated with a significant increase in the risk of cardiac events (dominated by recurrent episodes of syncope) in LQT2 women, suggesting that careful follow-up and continued long-term therapy are warranted in this population. PMID- 21632496 TI - Impact of leukocyte count on mortality and bleeding in patients with myocardial infarction undergoing primary percutaneous coronary interventions: analysis from the Harmonizing Outcome with Revascularization and Stent in Acute Myocardial Infarction trial. AB - BACKGROUND: The relationship between white blood cell count (WBCc) and mortality in patients with ST-segment-elevation acute myocardial infarction treated with percutaneous coronary intervention is poorly understood. Furthermore, whether there is a relationship between WBCc and risk of noncardiac mortality and bleeding after percutaneous coronary intervention is unknown. METHODS AND RESULTS: The baseline WBCc was available in 3193 of 3345 patients (95.5%) who underwent percutaneous coronary intervention in the Harmonizing Outcome With Revascularization and Stent in Acute Myocardial Infarction (HORIZONS-AMI) trial. In a propensity-adjusted multivariable analysis, WBCc was an independent predictor of 1-year cardiac mortality (hazard ratio, 1.15; 95% confidence interval, 1.09 to 1.22), noncardiac mortality (hazard ratio, 1.19; 95% confidence interval, 1.10 to 1.29), and major bleeding (hazard ratio, 1.08; 95% confidence interval, 1.04 to 1.12). After adjustment for baseline creatinine phosphokinase levels and left ventricular ejection fraction, WBCc remained an independent predictor of 1-year all-cause mortality and cardiac mortality. In patients matched for baseline creatinine phosphokinase levels at hospital admission, the median peak creatinine phosphokinase level was significantly higher in patients with high WBCc (>11 000 per 1 mm(3)) compared with low WBCc (1851 U/L [range, 880 3307 U/L] versus 1241 U/L [range, 540 to 2,78], respectively; P<0.0001). In this subgroup of patients, WBCc was an independent correlate of peak creatinine phosphokinase level, and remained an independent predictor of 1-year mortality. CONCLUSIONS: In patients with ST-segment-elevation acute myocardial infarction undergoing percutaneous coronary intervention, elevated baseline WBCc is an independent predictor of infarct size, as assessed by peak creatinine phosphokinase level, and of 1-year cardiac mortality, noncardiac mortality, and major bleeding. PMID- 21632497 TI - Blood pressure targets in subjects with type 2 diabetes mellitus/impaired fasting glucose: observations from traditional and bayesian random-effects meta-analyses of randomized trials. AB - BACKGROUND: Most guidelines for treatment of hypertension recommend a blood pressure (BP) goal of <140/90 mm Hg, and a more aggressive goal of <130/80 mm Hg for patients with diabetes mellitus. However, in the recent Action to Control Cardiovascular Risk in Diabetes (ACCORD) trial, a lower BP was not beneficial. The optimal BP target in subjects with diabetes mellitus or those with impaired fasting glucose/glucose tolerance is therefore not well defined. METHODS AND RESULTS: We performed PUBMED, EMBASE, and CENTRAL searches for randomized clinical trials from 1965 through October 2010 of antihypertensive therapy in patients with type 2 diabetes mellitus or impaired fasting glucose/impaired glucose tolerance that enrolled at least 100 patients with achieved systolic BP of <= 135 mm Hg in the intensive BP control group and <= 140 mm Hg in the standard BP control group, had a follow-up of at least 1 year, and evaluated macrovascular or microvascular events. We identified 13 randomized clinical trials enrolling 37 736 participants. Intensive BP control was associated with a 10% reduction in all-cause mortality (odds ratio, 0.90; 95% confidence interval, 0.83 to 0.98), a 17% reduction in stroke, and a 20% increase in serious adverse effects, but with similar outcomes for other macrovascular and microvascular (cardiac, renal, and retinal) events compared with standard BP control. The results were similar in a sensitivity analysis using a bayesian random-effects model. More intensive BP control (<= 130 mm Hg) was associated with a greater reduction in stroke, but did not reduce other events. Meta-regression analysis showed continued risk reduction for stroke to a systolic BP of <120 mm Hg. However, at levels <130 mm Hg, there was a 40% increase in serious adverse events with no benefit for other outcomes. CONCLUSIONS: The present body of evidence suggests that in patients with type 2 diabetes mellitus/impaired fasting glucose/impaired glucose tolerance, a systolic BP treatment goal of 130 to 135 mm Hg is acceptable. However, with more aggressive goals (<130 mm Hg), we observed target organ heterogeneity in that the risk of stroke continued to fall, but there was no benefit regarding the risk of other macrovascular or microvascular (cardiac, renal and retinal) events, and the risk of serious adverse events even increased. PMID- 21632498 TI - Long-term prognosis of acute myeloid leukemia according to the new genetic risk classification of the European LeukemiaNet recommendations: evaluation of the proposed reporting system. AB - PURPOSE: The current European LeukemiaNet (ELN) recommendations for acute myeloid leukemia (AML) propose a new risk reporting system, integrating molecular and cytogenetic factors and subdividing the large heterogenous group of intermediate risk patients into intermediate-I (IR-I) and intermediate-II (IR-II). We assessed the prognostic value of the new risk classification in a large cohort of patients. PATIENTS AND METHODS: Complete data for classification were available for 1,557 of 1,862 patients treated in the AML96 trial. Patients were assigned to the proposed genetic groups from the ELN recommendations, and survival analyses were performed using the Kaplan-Meier method and log-rank test for significance testing. RESULTS: The median age of all patients was 67 years. With a median follow-up of 8.3 years, significant differences between all risk categories were observed in patients age <= 60 years regarding the time to relapse, relapse-free survival, and overall survival (OS). Patients in the IR-II group had a better prognosis than patients in the IR-I group. The median OS times in young patients with favorable risk (FR), IR-I, IR-II, and adverse risk (AR) were 5.3, 1.1, 1.6, and 0.5 years, respectively. Separate analyses in the age group older than 60 years revealed significant differences between FR, AR, and IR as a whole, but not between IR-I and IR-II. CONCLUSION: In younger patients with AML, the ELN classification seems to be the best available framework for prognostic estimations to date. Caution is advised concerning its use for prospective treatment allocation before it has been prospectively validated. In elderly patients, alternative prognostic factors are desirable for further risk stratification of IR. PMID- 21632499 TI - Identifying patients with follicular lymphoma who are likely to benefit from an idiotype vaccine. PMID- 21632500 TI - Expanding our therapeutic options: Beta blockers for breast cancer? PMID- 21632501 TI - Beta-blocker use is associated with improved relapse-free survival in patients with triple-negative breast cancer. AB - PURPOSE: To examine the association between beta-blocker (BB) intake, pathologic complete response (pCR) rates, and survival outcomes in patients with breast cancer treated with neoadjuvant chemotherapy. PATIENTS AND METHODS: We retrospectively reviewed 1,413 patients with breast cancer who received neoadjuvant chemotherapy between 1995 and 2007. Patients taking BBs at the start of neoadjuvant therapy were compared with patients with no BB intake. Rates of pCR between the groups were compared using a chi2 test. Cox proportional hazards models were fitted to determine the association between BB intake, relapse-free survival (RFS), and overall survival (OS). RESULTS: Patients who used BBs (n = 102) were compared with patients (n = 1,311) who did not. Patients receiving BBs tended to be older and obese (P < .001). The proportion of pCR was not significantly different between the groups (P = .48). After adjustment for age, race, stage, grade, receptor status, lymphovascular invasion, body mass index, diabetes, hypertension, and angiotensin-converting enzyme inhibitor use, BB intake was associated with a significantly better RFS (hazard ratio [HR], 0.52; 95% CI, 0.31 to 0.88) but not OS (P = .09). Among patients with triple-negative breast cancer (TNBC; n = 377), BB intake was associated with improved RFS (HR, 0.30; 95% CI, 0.10 to 0.87;P = .027) but not OS (HR, 0.35; 95% CI, 0.12 to 1.00;P = .05). CONCLUSION: In this study, BB intake was associated with improved RFS in all patients with breast cancer and in patients with TNBC. Additional studies evaluating the potential benefits of beta-adrenergic blockade on breast cancer recurrence with a focus on TNBC are warranted. PMID- 21632502 TI - Pathology review of outside material: when does it help and when can it hurt? AB - PURPOSE: Pathology review is performed for patients when care is transferred to a tertiary care center after diagnostic tissue has been obtained. While it has many benefits, this practice can lead to unforeseen difficulties in doctor-patient communication and patient well-being, especially if a diagnosis is overturned or modified years after treatment. The aim of this analysis is to identify clinical situations in which pathology review can result in challenging discussions between patients and oncologists. PATIENTS: Representative case scenarios are presented in the subspecialty area of hematopathology. Analysis of the clinical benefits and possible harm to patients, pathologists, and treating oncologists that may ensue from pathology review is performed. RESULTS: Pathology review may result in a valuable second opinion and expert subclassification. However, problematic situations may arise with pathology review, especially if the patient has already undergone definitive treatment and is referred to an academic institution in remission. Difficulties can also arise when patients do not understand the limitations of diagnosing disease on small biopsies. The patient may receive a different diagnosis or it may become apparent that the diagnosis could have been made more expeditiously. These discrepancies must be communicated to the patient and may cause confusion and distress. CONCLUSION: Pathology review can be beneficial or potentially harmful depending on the clinical situation. Preliminary recommendations are provided for selecting patients for review. Limiting pathology review to certain clinical situations and encouraging patients to get second opinions before initial treatment at local referral centers may be helpful in minimizing reassignment of diagnoses after definitive treatment. PMID- 21632503 TI - Beta blockers and breast cancer mortality: a population- based study. AB - PURPOSE: Preclinical studies have demonstrated that antagonism of beta2 adrenergic signaling inhibits several pathways necessary for breast tumor progression and metastasis. A series of population-based observational studies were conducted to examine associations between beta blocker use and breast tumor characteristics at diagnosis or breast cancer-specific mortality. PATIENTS AND METHODS: Linked national cancer registry and prescription dispensing data were used to identify women with a diagnosis of stage I to IV invasive breast cancer between January 1, 2001, and December 31, 2006. Women taking propranolol (beta1/beta2 antagonist; n = 70) or atenolol (beta1 antagonist; n = 525), in the year before breast cancer diagnosis were matched (1:2) to women not taking a beta blocker (n = 4,738). Associations between use of propranolol or atenolol and risk of local tumor invasion at diagnosis (T4 tumor), nodal or metastatic involvement at diagnosis (N2/N3/M1 tumor), and time to breast cancer-specific mortality were assessed. RESULTS: Propranolol users were significantly less likely to present with a T4 (odds ratio [OR], 0.24, 95% CI, 0.07 to 0.85) or N2/N3/M1 (OR, 0.20; 95% CI, 0.04 to 0.88) tumor compared with matched nonusers. The cumulative probability of breast cancer-specific mortality was significantly lower for propranolol users compared with matched nonusers (hazard ratio, 0.19; 95% CI, 0.06 to 0.60). There was no difference in T4 or N2/N3/M1 tumor incidence or breast cancer-specific mortality between atenolol users and matched nonusers. CONCLUSION: The results provide evidence in humans to support preclinical observations suggesting that inhibiting the beta2-adrenergic signaling pathway can reduce breast cancer progression and mortality. PMID- 21632504 TI - Vaccination with patient-specific tumor-derived antigen in first remission improves disease-free survival in follicular lymphoma. AB - PURPOSE: Vaccination with hybridoma-derived autologous tumor immunoglobulin (Ig) idiotype (Id) conjugated to keyhole limpet hemocyanin (KLH) and administered with granulocyte-monocyte colony-stimulating factor (GM-CSF) induces follicular lymphoma (FL) -specific immune responses. To determine the clinical benefit of this vaccine, we conducted a double-blind multicenter controlled phase III trial. PATIENTS AND METHODS: Treatment-naive patients with advanced stage FL achieving complete response (CR) or CR unconfirmed (CRu) after chemotherapy were randomly assigned two to one to receive either Id vaccine (Id-KLH + GM-CSF) or control (KLH + GM-CSF). Primary efficacy end points were disease-free survival (DFS) for all randomly assigned patients and DFS for randomly assigned patients receiving at least one dose of Id vaccine or control. RESULTS: Of 234 patients enrolled, 177 (81%) achieved CR/CRu after chemotherapy and were randomly assigned. For 177 randomly assigned patients, including 60 patients not vaccinated because of relapse (n = 55) or other reasons (n = 5), median DFS between Id-vaccine and control arms was 23.0 versus 20.6 months, respectively (hazard ratio [HR], 0.81; 95% CI, 0.56 to 1.16; P = .256). For 117 patients who received Id vaccine (n = 76) or control (n = 41), median DFS after randomization was 44.2 months for Id vaccine arm versus 30.6 months for control arm (HR, 0.62; 95% CI, 0.39 to 0.99; P = .047) at median follow-up of 56.6 months (range, 12.6 to 89.3 months). In an unplanned subgroup analysis, median DFS was significantly prolonged for patients receiving IgM-Id (52.9 v 28.7 months; P = .001) but not IgG-Id vaccine (35.1 v 32.4 months; P = .807) compared with isotype-matched control-treated patients. CONCLUSION: Vaccination with patient-specific hybridoma-derived Id vaccine after chemotherapy-induced CR/CRu may prolong DFS in patients with FL. Vaccine isotype may affect clinical outcome and explain differing results between this and other controlled Id-vaccine trials. PMID- 21632505 TI - Adult medulloblastoma comprises three major molecular variants. AB - PURPOSE: Medulloblastoma is a rare primary brain tumor in adults, whereas it constitutes the most common malignant brain tumor in children. Integrated genomics approaches revealed at least four distinct disease variants in children. The aim of this study was to investigate molecular subtypes and their prognostic implication in a large cohort of adult medulloblastomas as the biology in this age group remains poorly understood. PATIENTS AND METHODS: We combined transcriptome and DNA copy number analyses for 28 adult medulloblastomas. Statistical and bioinformatic tools were applied to discover distinct molecular variants. Clinical and molecular characteristics of each biologic subtype were validated using immunohistochemistry on a tissue microarray derived from an independent patient cohort of adult medulloblastomas (n = 103). RESULTS: Gene expression profiles revealed three distinct molecular variants with stable subtype separation using the 300 most varying transcripts. Distinct demographics, genetics, transcriptome, and prognosis were noted for each subtype of adult medulloblastoma. Immunohistochemistry revealed aberrant activation of the sonic hedgehog (SHH) pathway in over half of adult medulloblastomas constituting a promising molecular therapeutic target. In contrast, subtype C tumors, which comprise a robust subtype in childhood medulloblastoma are only exceptionally seen in adult cohorts. Notably, adult subtype D and Wnt/wingless tumors were associated with worse prognosis than pediatric cohorts, whereas survival for SHH tumors was similar for both age groups. CONCLUSION: The transcriptome of adult medulloblastomas differs considerably from pediatric counterparts, both in terms of tumor biology and prognostic impact. Therefore, age-specific classification is required and must be adapted for use in clinical trials of adult medulloblastoma. PMID- 21632506 TI - Proof of activity of anti-epidermal growth factor receptor-targeted therapy for relapsed squamous cell carcinoma of the penis. PMID- 21632507 TI - Clear cell adenocarcinoma with enteroblastic differentiation of the ascending colon. PMID- 21632508 TI - National estimates of out-of-pocket health care expenditure burdens among nonelderly adults with cancer: 2001 to 2008. AB - PURPOSE: To compare the prevalence of high out-of-pocket burdens among patients with cancer with other chronically ill and well patients, and to examine the sociodemographic characteristics associated with high burdens among patients with cancer. METHODS: The sample included persons 18 to 64 years of age who received treatment for cancer, taken from a nationally representative sample of the US population from the 2001 to 2008 Medical Expenditure Panel Survey. We examined the proportion of persons living in families with high out-of-pocket burdens associated with medical spending, including insurance premiums, relative to income, defining high health care (total) burden as spending more than 20% of income on health care (and premiums). RESULTS: The risk of high burdens is significantly greater for patients with cancer compared with other chronically ill and well patients. We find that 13.4% of patients with cancer had high total burdens, in contrast to 9.7% among those with other chronic conditions and 4.4% among those without chronic conditions. Among nonelderly persons with cancer, the following were associated with higher out-of-pocket burdens: private nongroup insurance, age 55 to 64 years, non-Hispanic black, never married or widowed, one child or no children, unemployed, lower income, lower education level, living in nonmetropolitan statistical areas, and having other chronic conditions. CONCLUSION: High burdens may affect treatment choice and deter patients from getting care. Thus, although a detailed patient-physician discussion of costs of care may not be feasible, we believe that an awareness of out-of-pocket burdens among patients with cancer is useful for clinical oncologists. PMID- 21632509 TI - Randomized phase III trial of paclitaxel/carboplatin with or without PF-3512676 (Toll-like receptor 9 agonist) as first-line treatment for advanced non-small cell lung cancer. AB - PURPOSE: This phase III study examined efficacy of the synthetic Toll-like receptor 9-activating oligodeoxynucleotide PF-3512676 in combination with standard paclitaxel/carboplatin chemotherapy in patients with advanced non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: Chemotherapy-naive patients with stage IIIB or IV NSCLC were randomly assigned (1:1) to receive up to six courses of paclitaxel/carboplatin (intravenous paclitaxel 200 mg/m(2) and carboplatin at area under the [concentration-time] curve 6 on day 1 of a 3-week cycle) alone (control arm) or in combination with 0.2 mg/kg subcutaneous PF-3512676 on days 8 and 15 (investigational arm). Primary end point was overall survival (OS). RESULTS: Baseline demographics were similar across arms (N = 828). Most patients (88%) had stage IV disease. Median OS and median progression-free survival (PFS) were similar (OS: investigational arm, 10.0 months v control arm, 9.8 months; P = .56; PFS: investigational arm, 4.8 months v control arm, 4.7 months; P = .79). Most commonly reported PF-3512676-related adverse events (AEs) were mild-to moderate local injection site reactions, pyrexia, and flu-like symptoms. In the investigational arm, grades 3 to 4 AEs, including neutropenia, thrombocytopenia, and anemia, were more frequent, and more patients had one or more sepsis-related AEs versus controls (17 v 3). At first interim analysis, the Data Safety Monitoring Committee recommended study discontinuation because of lack of incremental efficacy and more sepsis-related serious AEs in the PF-3512676 arm. Administration of PF-3512676, but not chemotherapy, was halted. CONCLUSION: Addition of PF-3512676 to paclitaxel/carboplatin did not improve OS or PFS versus paclitaxel/carboplatin alone for first-line treatment of patients with advanced NSCLC but did increase toxicity. This regimen cannot be recommended for treating patients with advanced NSCLC. PMID- 21632510 TI - Progression-free survival in neuroendocrine tumors: preferred end point, but how should it be defined? PMID- 21632511 TI - Changes in prostate cancer grade on serial biopsy in men undergoing active surveillance. AB - PURPOSE: Active surveillance is now considered a viable treatment option for men with low-risk prostate cancer. However, little is known regarding changes in Gleason grade on serial biopsies over an extended period of time. PATIENTS AND METHODS: Men diagnosed with prostate cancer between 1998 and 2009 who elected active surveillance as initial treatment, with 6 or more months of follow-up and a minimum of six cores at biopsy, were included in analysis. Upgrading and downgrading were defined as an increase or decrease in primary or secondary Gleason score. Means and frequency tables were used to describe patient characteristics, and treatment-free survival rates were determined by life-table product limit estimates. RESULTS: Three hundred seventy-seven men met inclusion criteria. Mean age at diagnosis was 61.9 years. Fifty-three percent of men had prostate-specific antigen of 6 ng/mL or less, and 94% had Gleason score of 6 or less. A majority of men were cT1 (62%), had less than 33% of biopsy cores involved (80%), and were low risk (77%) at diagnosis. Median number of cores taken at diagnostic biopsy was 13, mean time to follow-up was 18.5 months, and 29% of men had three or more repeat biopsies. Overall, 34% (129 men) were found to have an increase in Gleason grade. The majority of men who experienced an upgrade (81%) did so by their second repeat biopsy. CONCLUSION: A proportion of men experience an upgrade in Gleason score while undergoing active surveillance. Men who experience early upgrading likely represent initial sampling error, whereas later upgrading may reflect tumor dedifferentiation. PMID- 21632512 TI - Cardiology patient page: cardiovascular implications of erectile dysfunction. PMID- 21632513 TI - Atrial septal defect with total anomalous pulmonary venous return in an adult. PMID- 21632514 TI - Letter by Montani et al regarding article, "Elevated levels of inflammatory cytokines predict survival in idiopathic and familial pulmonary arterial hypertension". PMID- 21632515 TI - Hydralazine and isosorbide dinitrate in heart failure: historical perspective, mechanisms, and future directions. PMID- 21632516 TI - Risk stratification for arrhythmic sudden cardiac death: identifying the roadblocks. PMID- 21632517 TI - Successful hybrid rescue of occluded pulmonary artery in pulmonary atresia. PMID- 21632518 TI - How longer saccade latencies lead to a competition for salience. AB - It has been suggested that independent bottom-up and top-down processes govern saccadic selection. However, recent findings are hard to explain in such terms. We hypothesized that differences in visual-processing time can explain these findings, and we tested this using search displays containing two deviating elements, one requiring a short processing time and one requiring a long processing time. Following short saccade latencies, the deviation requiring less processing time was selected most frequently. This bias disappeared following long saccade latencies. Our results suggest that an element that attracts eye movements following short saccade latencies does so because it is the only element processed at that time. The temporal constraints of processing visual information therefore seem to be a determining factor in saccadic selection. Thus, relative saliency is a time-dependent phenomenon. PMID- 21632519 TI - Allergy among veterinary medicine students in The Netherlands. AB - BACKGROUND: Veterinary medicine students who practice with animals are potentially exposed to many occupational agents, yet sensitisation and allergic symptoms among this group have not been studied extensively. OBJECTIVE: The objective of this study was to estimate the prevalence of sensitisation and allergic symptoms in veterinary medicine students in association with study specialisation over time. METHODS: A questionnaire-based cross-sectional study was conducted. Blood was collected and tested for total and specific serum IgE for 16 different common and study-specific allergens using enzyme immunoassay. RESULTS: New development of self-reported allergic symptoms to various allergens occurred in 8.7%, of which 44% was deducted against animals. Handling farm animals was strongly associated with self-reported allergies to various allergens (OR=6.9, 95% CI 1.9 to 25) and animal allergens (OR=12, 95% CI 1.4 to 103). Sensitisation to at least one allergen occurred in 33.1%. Sensitisation prevalence tended to be elevated in later years of the equine study program. In contrast to self-reported allergies, the prevalence of sensitisation to any allergen decreased with prolonged study duration for those specialising in farm animal health (years 3-5: OR=0.5, 95% CI 0.3 to 1.1; year 6: OR=0.2, 95% CI 0.1 to 0.5). This was independent of whether people were raised on a farm, which is in itself a protective factor for allergy and sensitisation. CONCLUSION: This study provides evidence of an elevated prevalence of allergic symptoms with increasing years of veterinary study, suggesting that contact with animals, more specifically contact to farm animals, is a risk factor for the development of symptoms. PMID- 21632520 TI - Cost-benefit analysis in occupational health: a comparison of intervention scenarios for occupational asthma and rhinitis among bakery workers. AB - OBJECTIVES: Use of cost-benefit analysis in occupational health increases insight into the intervention strategy that maximises the cost-benefit ratio. This study presents a methodological framework identifying the most important elements of a cost-benefit analysis for occupational health settings. One of the main aims of the methodology is to evaluate cost-benefit ratios for different stakeholders (employers, employees and society). The developed methodology was applied to two intervention strategies focused on reducing respiratory diseases. METHODS: A cost benefit framework was developed and used to set up a calculation spreadsheet containing the inputs and algorithms required to calculate the costs and benefits for all cost elements. Inputs from a large variety of sources were used to calculate total costs, total benefits, net costs and the benefit-to-costs ratio for both intervention scenarios. RESULTS: Implementation of a covenant intervention program resulted in a net benefit of ?16 848 546 over 20 years for a population of 10 000 workers. Implementation was cost-effective for all stakeholders. For a health surveillance scenario, total benefits resulting from a decreased disease burden were estimated to be ?44 659 352. The costs of the interventions could not be calculated. CONCLUSION: This study provides important insights for developing effective intervention strategies in the field of occupational medicine. Use of a model based approach enables investigation of those parameters most likely to impact on the effectiveness and costs of interventions for work related diseases. Our case study highlights the importance of considering different perspectives (of employers, society and employees) in assessing and sharing the costs and benefits of interventions. PMID- 21632521 TI - Child injury prevention and child survival. PMID- 21632522 TI - High prevalence of subclinical tuberculosis in HIV-1-infected persons without advanced immunodeficiency: implications for TB screening. AB - BACKGROUND: The prevalence of asymptomatic tuberculosis (TB) in recently diagnosed HIV-1-infected persons attending pre-antiretroviral therapy (ART) clinics is not well described. In addition, it is unclear if the detection of Mycobacterium tuberculosis in these patients clearly represents an early asymptomatic phase leading to progressive disease or transient excretion of bacilli. OBJECTIVE: To describe the prevalence and outcome of subclinical TB disease in HIV-1-infected persons not eligible for ART. METHODS: The study was conducted in 274 asymptomatic ART-naive HIV-1-infected persons in Khayelitsha Day Hospital, Cape Town, South Africa. All participants were screened for TB using a symptom screen and spoligotyping was performed to determine genotypes. RESULTS: The prevalence of subclinical TB disease was 8.5% (95% CI 5.1% to 13.0%) (n=18; median days to culture positivity 17 days), with 22% of patients being smear-positive. Spoligotyping showed a diverse variety of genotypes with all paired isolates being of the same spoligotype, effectively excluding cross contamination. 56% of patients followed up developed symptoms 3 days to 2 months later. All were well and still in care 6-12 months after TB diagnosis; 60% were started on ART. A positive tuberculin skin test (OR 4.96, p=0.064), low CD4 count (OR 0.996, p=0.06) and number of years since HIV diagnosis (OR 1.006, p=0.056) showed trends towards predicting TB disease. Conclusion This study found a high prevalence but good outcome (retained in care) of subclinical TB disease in HIV-1 infected persons. The results suggest that, in high HIV/TB endemic settings, a positive HIV-1 test should prompt TB screening by sputum culture irrespective of symptoms, particularly in those with a positive tuberculin skin test, longer history of HIV infection and low CD4 count. Operational difficulties in resource constrained settings with respect to screening with TB culture highlight the need for rapid and affordable point-of-care tests to identify persons with clinical and subclinical TB disease. PMID- 21632523 TI - A role for XRCC2 gene polymorphisms in breast cancer risk and survival. AB - BACKGROUND: The XRCC2 gene is a key mediator in the homologous recombination repair of DNA double strand breaks. It is hypothesised that inherited variants in the XRCC2 gene might also affect susceptibility to, and survival from, breast cancer. METHODS: The study genotyped 12 XRCC2 tagging single nucleotide polymorphisms (SNPs) in 1131 breast cancer cases and 1148 controls from the Sheffield Breast Cancer Study (SBCS), and examined their associations with breast cancer risk and survival by estimating ORs and HRs, and their corresponding 95% CIs. Positive findings were further investigated in 860 cases and 869 controls from the Utah Breast Cancer Study (UBCS) and jointly analysed together with available published data for breast cancer risk. The survival findings were further confirmed in studies (8074 cases) from the Breast Cancer Association Consortium (BCAC). RESULTS: The most significant association with breast cancer risk in the SBCS dataset was the XRCC2 rs3218408 SNP (recessive model p=2.3*10( 4), minor allele frequency (MAF)=0.23). This SNP yielded an OR(rec) of 1.64 (95% CI 1.25 to 2.16) in a two-site analysis of SBCS and UBCS, and a meta-OR(rec) of 1.33 (95% CI 1.12 to 1.57) when all published data were included. This SNP may mark a rare risk haplotype carried by two in 1000 of the control population. Furthermore, the XRCC2 coding R188H SNP (rs3218536, MAF=0.08) was significantly associated with poor survival, with an increased per-allele HR of 1.58 (95% CI 1.01 to 2.49) in a multivariate analysis. This effect was still evident in a pooled meta-analysis of 8781 breast cancer patients from the BCAC (HR 1.19, 95% CI 1.05 to 1.36; p=0.01). CONCLUSIONS: These findings suggest that XRCC2 SNPs may influence breast cancer risk and survival. PMID- 21632524 TI - Cool head, hot brain: cerebral blood flow distribution during exercise. PMID- 21632525 TI - Role of neuronal potassium M-channels in sympathetic regulation of cardiac function. PMID- 21632526 TI - Spinal breathing: stimulation and surprises. PMID- 21632527 TI - Measuring calcium in 'fuzzy' spaces. PMID- 21632528 TI - Skeletal muscle stiffness and contracture in children with spastic cerebral palsy. PMID- 21632529 TI - No role for early IGF-1 signalling in stimulating acute 'muscle building' responses. PMID- 21632530 TI - Crystal structure of the heme d1 biosynthesis enzyme NirE in complex with its substrate reveals new insights into the catalytic mechanism of S-adenosyl-L methionine-dependent uroporphyrinogen III methyltransferases. AB - During the biosynthesis of heme d(1), the essential cofactor of cytochrome cd(1) nitrite reductase, the NirE protein catalyzes the methylation of uroporphyrinogen III to precorrin-2 using S-adenosyl-L-methionine (SAM) as the methyl group donor. The crystal structure of Pseudomonas aeruginosa NirE in complex with its substrate uroporphyrinogen III and the reaction by-product S-adenosyl-L homocysteine (SAH) was solved to 2.0 A resolution. This represents the first enzyme-substrate complex structure for a SAM-dependent uroporphyrinogen III methyltransferase. The large substrate binds on top of the SAH in a "puckered" conformation in which the two pyrrole rings facing each other point into the same direction either upward or downward. Three arginine residues, a histidine, and a methionine are involved in the coordination of uroporphyrinogen III. Through site directed mutagenesis of the nirE gene and biochemical characterization of the corresponding NirE variants the amino acid residues Arg-111, Glu-114, and Arg-149 were identified to be involved in NirE catalysis. Based on our structural and biochemical findings, we propose a potential catalytic mechanism for NirE in which the methyl transfer reaction is initiated by an arginine catalyzed proton abstraction from the C-20 position of the substrate. PMID- 21632531 TI - Substrate preferences and catalytic parameters determined by structural characteristics of sterol 14alpha-demethylase (CYP51) from Leishmania infantum. AB - Leishmaniasis is a major health problem that affects populations of ~90 countries worldwide, with no vaccine and only a few moderately effective drugs. Here we report the structure/function characterization of sterol 14alpha-demethylase (CYP51) from Leishmania infantum. The enzyme catalyzes removal of the 14alpha methyl group from sterol precursors. The reaction is essential for membrane biogenesis and therefore has great potential to become a target for antileishmanial chemotherapy. Although L. infantum CYP51 prefers C4 monomethylated sterol substrates such as C4-norlanosterol and obtusifoliol (V(max) of ~10 and 8 min(-1), respectively), it is also found to 14alpha demethylate C4-dimethylated lanosterol (V(max) = 0.9 min(-1)) and C4 desmethylated 14alpha-methylzymosterol (V(max) = 1.9 min(-1)). Binding parameters with six sterols were tested, with K(d) values ranging from 0.25 to 1.4 MUM. Thus, L. infantum CYP51 is the first example of a plant-like sterol 14alpha demethylase, where requirements toward the composition of the C4 atom substituents are not strict, indicative of possible branching in the postsqualene portion of sterol biosynthesis in the parasite. Comparative analysis of three CYP51 substrate binding cavities (Trypanosoma brucei, Trypanosoma cruzi, and L. infantum) suggests that substrate preferences of plant- and fungal-like protozoan CYP51s largely depend on the differences in the enzyme active site topology. These minor structural differences are also likely to underlie CYP51 catalytic rates and drug susceptibility and can be used to design potent and specific inhibitors. PMID- 21632532 TI - Liquid chromatography-mass spectrometry-based quantitative proteomics. AB - LC-MS-based quantitative proteomics has become increasingly applied to a wide range of biological applications due to growing capabilities for broad proteome coverage and good accuracy and precision in quantification. Herein, we review the current LC-MS-based quantification methods with respect to their advantages and limitations and highlight their potential applications. PMID- 21632533 TI - Lowering bile acid pool size with a synthetic farnesoid X receptor (FXR) agonist induces obesity and diabetes through reduced energy expenditure. AB - We evaluated the metabolic impact of farnesoid X receptor (FXR) activation by administering a synthetic FXR agonist (GW4064) to mice in which obesity was induced by a high fat diet. Administration of GW4064 accentuated body weight gain and glucose intolerance induced by the high fat diet and led to a pronounced worsening of the changes in liver and adipose tissue. Mechanistically, treatment with GW4064 decreased bile acid (BA) biosynthesis, BA pool size, and energy expenditure, whereas reconstitution of the BA pool in these GW4064-treated animals by BA administration dose-dependently reverted the metabolic abnormalities. Our data therefore suggest that activation of FXR with synthetic agonists is not useful for long term management of the metabolic syndrome, as it reduces the BA pool size and subsequently decreases energy expenditure, translating as weight gain and insulin resistance. In contrast, expansion of the BA pool size, which can be achieved by BA administration, could be an interesting strategy to manage the metabolic syndrome. PMID- 21632534 TI - Ubiquitin chain trimming recycles the substrate binding sites of the 26 S proteasome and promotes degradation of lysine 48-linked polyubiquitin conjugates. AB - The 26 S proteasome possesses two distinct deubiquitinating activities. The ubiquitin (Ub) chain amputation activity removes the entire polyUb chain from the substrates. The Ub chain trimming activity progressively cleaves a polyUb chain from the distal end. The Ub chain amputation activity mediates degradation coupled deubiquitination. The Ub chain trimming activity can play a supportive or an inhibitory role in degradation, likely depending on features of the substrates. How Ub chain trimming assists degradation is not clear. We find that inhibition of the chain trimming activity of the 26 S proteasome with Ub aldehyde significantly inhibits degradation of Ub4 (Lys-48)-UbcH10 and causes accumulation of free Ub4 (generated from chain amputation) that can be retained on the proteasome. Also, a non-trimmable Lys-48-mimic Ub4 efficiently targets UbcH10 to the 26 S proteasome, but it cannot support efficient degradation of UbcH10 compared with regular Lys-48 Ub4. These results indicate that polyUb chain trimming promotes proteasomal degradation of Lys-48-linked substrates. Mechanistically, we propose that Ub chain trimming cleaves the proteasome-bound Lys-48-linked polyUb chains, which vacates the Ub binding sites of the 26 S proteasome, thus allowing continuous substrate loading. PMID- 21632536 TI - 3-Keto-5-aminohexanoate cleavage enzyme: a common fold for an uncommon Claisen type condensation. AB - The exponential increase in genome sequencing output has led to the accumulation of thousands of predicted genes lacking a proper functional annotation. Among this mass of hypothetical proteins, enzymes catalyzing new reactions or using novel ways to catalyze already known reactions might still wait to be identified. Here, we provide a structural and biochemical characterization of the 3-keto-5 aminohexanoate cleavage enzyme (Kce), an enzymatic activity long known as being involved in the anaerobic fermentation of lysine but whose catalytic mechanism has remained elusive so far. Although the enzyme shows the ubiquitous triose phosphate isomerase (TIM) barrel fold and a Zn(2+) cation reminiscent of metal dependent class II aldolases, our results based on a combination of x-ray snapshots and molecular modeling point to an unprecedented mechanism that proceeds through deprotonation of the 3-keto-5-aminohexanoate substrate, nucleophilic addition onto an incoming acetyl-CoA, intramolecular transfer of the CoA moiety, and final retro-Claisen reaction leading to acetoacetate and 3 aminobutyryl-CoA. This model also accounts for earlier observations showing the origin of carbon atoms in the products, as well as the absence of detection of any covalent acyl-enzyme intermediate. Kce is the first representative of a large family of prokaryotic hypothetical proteins, currently annotated as the "domain of unknown function" DUF849. PMID- 21632535 TI - Phosphodiesterase 3A (PDE3A) deletion suppresses proliferation of cultured murine vascular smooth muscle cells (VSMCs) via inhibition of mitogen-activated protein kinase (MAPK) signaling and alterations in critical cell cycle regulatory proteins. AB - Cyclic nucleotide phosphodiesterase 3 (PDE3) is an important regulator of cyclic adenosine monophosphate (cAMP) signaling within the cardiovascular system. In this study, we examined the role of PDE3A and PDE3B isoforms in regulation of growth of cultured vascular smooth muscle cells (VSMCs) and the mechanisms by which they may affect signaling pathways that mediate mitogen-induced VSMC proliferation. Serum- and PDGF-induced DNA synthesis in VSMCs grown from aortas of PDE3A-deficient (3A-KO) mice was markedly less than that in VSMCs from PDE3A wild type (3A-WT) and PDE3B-deficient (3B-KO) mice. The reduced growth response was accompanied by significantly less phosphorylation of extracellular signal regulated kinase (ERK) in 3A-KO VSMCs, most likely due to a combination of greater site-specific inhibitory phosphorylation of Raf-1(Ser-259) by protein kinase A (PKA) and enhanced dephosphorylation of ERKs due to elevated mitogen activated protein kinase phosphatase 1 (MKP-1). Furthermore, 3A-KO VSMCs, compared with 3A-WT, exhibited higher basal PKA activity and cAMP response element-binding protein (CREB) phosphorylation, higher levels of p53 and p53 phosphorylation, and elevated p21 protein together with lower levels of Cyclin-D1 and retinoblastoma (Rb) protein and Rb phosphorylation. Adenoviral overexpression of inactive CREB partially restored growth effects of serum in 3A-KO VSMCs. In contrast, exposure of 3A-WT VSMCs to VP16 CREB (active CREB) was associated with inhibition of serum-induced DNA synthesis similar to that in untreated 3A-KO VSMCs. Transfection of 3A-KO VSMCs with p53 siRNA reduced p21 and MKP-1 levels and completely restored growth without affecting amounts of Cyclin-D1 and Rb phosphorylation. We conclude that PDE3A regulates VSMC growth via two complementary pathways, i.e. PKA-catalyzed inhibitory phosphorylation of Raf-1 with resulting inhibition of MAPK signaling and PKA/CREB-mediated induction of p21, leading to G0/G1 cell cycle arrest, as well as by increased accumulation of p53, which induces MKP-1, p21, and WIP1, leading to inhibition of G1 to S cell cycle progression. PMID- 21632537 TI - Structure and mechanism of the diiron benzoyl-coenzyme A epoxidase BoxB. AB - The coenzyme A (CoA)-dependent aerobic benzoate metabolic pathway uses an unprecedented chemical strategy to overcome the high aromatic resonance energy by forming the non-aromatic 2,3-epoxybenzoyl-CoA. The crucial dearomatizing reaction is catalyzed by three enzymes, BoxABC, where BoxA is an NADPH-dependent reductase, BoxB is a benzoyl-CoA 2,3-epoxidase, and BoxC is an epoxide ring hydrolase. We characterized the key enzyme BoxB from Azoarcus evansii by structural and Mossbauer spectroscopic methods as a new member of class I diiron enzymes. Several family members were structurally studied with respect to the diiron center architecture, but no structure of an intact diiron enzyme with its natural substrate has been reported. X-ray structures between 1.9 and 2.5 A resolution were determined for BoxB in the diferric state and with bound substrate benzoyl-CoA in the reduced state. The substrate-bound reduced state is distinguished from the diferric state by increased iron-ligand distances and the absence of directly bridging groups between them. The position of benzoyl-CoA inside a 20 A long channel and the position of the phenyl ring relative to the diiron center are accurately defined. The C2 and C3 atoms of the phenyl ring are closer to one of the irons. Therefore, one oxygen of activated O(2) must be ligated predominantly to this proximate iron to be in a geometrically suitable position to attack the phenyl ring. Consistent with the observed iron/phenyl geometry, BoxB stereoselectively should form the 2S,3R-epoxide. We postulate a reaction cycle that allows a charge delocalization because of the phenyl ring and the electron-withdrawing CoA thioester. PMID- 21632538 TI - Plant pathogenic bacteria utilize biofilm growth-associated repressor (BigR), a novel winged-helix redox switch, to control hydrogen sulfide detoxification under hypoxia. AB - Winged-helix transcriptional factors play important roles in the control of gene expression in many organisms. In the plant pathogens Xylella fastidiosa and Agrobacterium tumefaciens, the winged-helix protein BigR, a member of the ArsR/SmtB family of metal sensors, regulates transcription of the bigR operon involved in bacterial biofilm growth. Previous studies showed that BigR represses transcription of its own operon through the occupation of the RNA polymerase binding site; however, the signals that modulate its activity and the biological function of its operon are still poorly understood. Here we show that although BigR is a homodimer similar to metal sensors, it functions as a novel redox switch that derepresses transcription upon oxidation. Crystal structures of reduced and oxidized BigR reveal that formation of a disulfide bridge involving two critical cysteines induces conformational changes in the dimer that remarkably alter the topography of the winged-helix DNA-binding interface, precluding DNA binding. This structural mechanism of DNA association-dissociation is novel among winged-helix factors. Moreover, we demonstrate that the bigR operon is required for hydrogen sulfide detoxification through the action of a sulfur dioxygenase (Blh) and sulfite exporter. As hydrogen sulfide strongly inhibits cytochrome c oxidase, it must be eliminated to allow aerobic growth under low oxygen tension, an environmental condition found in bacterial biofilms, xylem vessels, and root tissues. Accordingly, we show that the bigR operon is critical to sustain bacterial growth under hypoxia. These results suggest that BigR integrates the transcriptional regulation of a sulfur oxidation pathway to an oxidative signal through a thiol-based redox switch. PMID- 21632539 TI - New applications of mass spectrometry in lipid analysis. AB - Mass spectrometry has emerged as a powerful tool for the analysis of all lipids. Lipidomic analysis of biological systems using various approaches is now possible with a quantitative measurement of hundreds of lipid molecular species. Although availability of reference and internal standards lags behind the field, approaches using stable isotope-labeled derivative tagging permit precise determination of specific phospholipids in an experimental series. The use of reactivity of ozone has enabled assessment of double bond positions in fatty acyl groups even when species remain in complex lipid mixtures. Rapid scanning tandem mass spectrometers are capable of quantitative analysis of hundreds of targeted lipids at high sensitivity in a single on-line chromatographic separation. Imaging mass spectrometry of lipids in tissues has opened new insights into the distribution of lipid molecular species with promising application to study pathophysiological events and diseases. PMID- 21632540 TI - Characterization of early EDEM1 protein maturation events and their functional implications. AB - The endoplasmic reticulum (ER) quality control factor EDEM1 associates with a number of ER proteins and ER-associated degradation (ERAD) substrates; however, an understanding of its role in ERAD is unclear. The early maturation events for EDEM1 including signal sequence cleavage and glycosylation were analyzed, and their relationship to the function of EDEM1 was determined. EDEM1 has five N linked glycosylation sites with the most C-terminal site recognized poorly cotranslationally, resulting in the accumulation of EDEM1 containing four or five glycans. The fifth site was modified post-translationally when bypassed cotranslationally. Signal sequence cleavage of EDEM1 was found to be a slow and inefficient process. Signal sequence cleavage produced a soluble form of EDEM1 that efficiently associated with the oxidoreductase ERdj5 and most effectively accelerated the turnover of a soluble ERAD substrate. In contrast, a type-II membrane form of EDEM1 was generated when the signal sequence was uncleaved, creating an N-terminal transmembrane segment. The membrane form of EDEM1 efficiently associated with the ER membrane protein SEL1L and accelerated the turnover of a membrane-associated ERAD substrate. Together, these results demonstrated that signal sequence cleavage functionally regulated the association of EDEM1-soluble and membrane-integrated isoforms with distinct ERAD machinery and substrates. PMID- 21632541 TI - Novel ganglioside-mediated entry of botulinum neurotoxin serotype D into neurons. AB - Botulinum Neurotoxins (BoNTs) are organized into seven serotypes, A-G. Although several BoNT serotypes enter neurons through synaptic vesicle cycling utilizing dual receptors (a ganglioside and a synaptic vesicle-associated protein), the entry pathway of BoNT/D is less well understood. Although BoNT/D entry is ganglioside-dependent, alignment and structural studies show that BoNT/D lacks key residues within a conserved ganglioside binding pocket that are present in BoNT serotypes A, B, E, F, and G, which indicate that BoNT/D-ganglioside interactions may be unique. In this study BoNT/D is shown to have a unique association with ganglioside relative to the other BoNT serotypes, utilizing a ganglioside binding loop (GBL, residues Tyr-1235-Ala-1245) within the receptor binding domain of BoNT/D (HCR/D) via b-series gangliosides, including GT1b, GD1b, and GD2. HCR/D bound gangliosides and entered neurons dependent upon the aromatic ring of Phe-1240 within the GBL. This is the first BoNT-ganglioside interaction that is mediated by a phenylalanine. In contrast, Trp-1238, located near the N terminus of the ganglioside binding loop, was mostly solvent-inaccessible and appeared to contribute to maintaining the loop structure. BoNT/D entry and intoxication were enhanced by membrane depolarization via synaptic vesicle cycling, where HCR/D colocalized with synaptophysin, a synaptic vesicle marker, but immunoprecipitation experiments did not detect direct association with synaptic vesicle protein 2. Thus, BoNT/D utilizes unique associations with gangliosides and synaptic vesicles to enter neurons, which may facilitate new neurotoxin therapies. PMID- 21632542 TI - Arabidopsis chloroplastic glutaredoxin C5 as a model to explore molecular determinants for iron-sulfur cluster binding into glutaredoxins. AB - Unlike thioredoxins, glutaredoxins are involved in iron-sulfur cluster assembly and in reduction of specific disulfides (i.e. protein-glutathione adducts), and thus they are also important redox regulators of chloroplast metabolism. Using GFP fusion, AtGrxC5 isoform, present exclusively in Brassicaceae, was shown to be localized in chloroplasts. A comparison of the biochemical, structural, and spectroscopic properties of Arabidopsis GrxC5 (WCSYC active site) with poplar GrxS12 (WCSYS active site), a chloroplastic paralog, indicated that, contrary to the solely apomonomeric GrxS12 isoform, AtGrxC5 exists as two forms when expressed in Escherichia coli. The monomeric apoprotein possesses deglutathionylation activity mediating the recycling of plastidial methionine sulfoxide reductase B1 and peroxiredoxin IIE, whereas the dimeric holoprotein incorporates a [2Fe-2S] cluster. Site-directed mutagenesis experiments and resolution of the x-ray crystal structure of AtGrxC5 in its holoform revealed that, although not involved in its ligation, the presence of the second active site cysteine (Cys(32)) is required for cluster formation. In addition, thiol titrations, fluorescence measurements, and mass spectrometry analyses showed that, despite the presence of a dithiol active site, AtGrxC5 does not form any inter- or intramolecular disulfide bond and that its activity exclusively relies on a monothiol mechanism. PMID- 21632543 TI - Mass spectrometry strategies in metabolomics. AB - MS has evolved as a critical component in metabolomics, which seeks to answer biological questions through large-scale qualitative and quantitative analyses of the metabolome. MS-based metabolomics techniques offer an excellent combination of sensitivity and selectivity, and they have become an indispensable platform in biology and metabolomics. In this minireview, various MS technologies used in metabolomics are briefly discussed, and future needs are suggested. PMID- 21632544 TI - Molecular recognition of leucine-aspartate repeat (LD) motifs by the focal adhesion targeting homology domain of cerebral cavernous malformation 3 (CCM3). AB - Cerebral cavernous malformation (CCM) is a disease that affects between 0.1 and 0.5% of the human population, with mutations in CCM3 accounting for ~ 15% of the autosomal dominant form of the disease. We recently reported that CCM3 contains an N-terminal dimerization domain (CCM3D) and a C-terminal focal adhesion targeting (FAT) homology domain. Intermolecular protein-protein interactions of CCM3 are mediated by a highly conserved surface on the FAT homology domain and are affected by CCM3 truncations in the human disease. Here we report the crystal structures of CCM3 in complex with three different leucine-aspartate repeat (LD) motifs (LD1, LD2, and LD4) from the scaffolding protein paxillin, at 2.8, 2.7, and 2.5 A resolution. We show that CCM3 binds LD motifs using the highly conserved hydrophobic patch 1 (HP1) and that this binding is similar to the binding of focal adhesion kinase and Pyk2 FAT domains to paxillin LD motifs. We further show by surface plasmon resonance that CCM3 binds paxillin LD motifs with affinities in the micromolar range, similar to FAK family FAT domains. Finally, we show that endogenous CCM3 and paxillin co-localize in mouse cerebral pericytes. These studies provide a molecular-level framework to investigate the protein-protein interactions of CCM3. PMID- 21632545 TI - Thematic minireview series on biological applications of mass spectrometry. AB - Mass spectrometry is a powerful technique with many applications in biology as well as chemistry and physics. The increases in sensitivity and resolution of the instruments, coupled with improvements in the analysis of data, have opened new dimensions in analyses of complex biological systems. Examples presented here include drug metabolism, lipid analysis, metabolomics, quantitative proteomics, direct analysis of intact proteins, and imaging of both small molecules and proteins in tissues. PMID- 21632546 TI - Drug metabolite profiling and identification by high-resolution mass spectrometry. AB - Mass spectrometry plays a key role in drug metabolite identification, an integral part of drug discovery and development. The development of high-resolution (HR) MS instrumentation with improved accuracy and stability, along with new data processing techniques, has improved the quality and productivity of metabolite identification processes. In this minireview, HR-MS-based targeted and non targeted acquisition methods and data mining techniques (e.g. mass defect, product ion, and isotope pattern filters and background subtraction) that facilitate metabolite identification are examined. Methods are presented that enable multiple metabolite identification tasks with a single LC/HR-MS platform and/or analysis. Also, application of HR-MS-based strategies to key metabolite identification activities and future developments in the field are discussed. PMID- 21632547 TI - The conserved Rieske oxygenase DAF-36/Neverland is a novel cholesterol metabolizing enzyme. AB - Steroid hormones play essential roles in a wide variety of biological processes in multicellular organisms. The principal steroid hormones in nematodes and arthropods are dafachronic acids and ecdysteroids, respectively, both of which are synthesized from cholesterol as an indispensable precursor. The first critical catalytic step in the biosynthesis of these ecdysozoan steroids is the conversion of cholesterol to 7-dehydrocholesterol. However, the enzymes responsible for cholesterol 7,8-dehydrogenation remain unclear at the molecular level. Here we report that the Rieske oxygenase DAF-36/Neverland (Nvd) is a cholesterol 7,8-dehydrogenase. The daf-36/nvd genes are evolutionarily conserved, not only in nematodes and insects but also in deuterostome species that do not produce dafachronic acids or ecdysteroids, including the sea urchin Hemicentrotus pulcherrimus, the sea squirt Ciona intestinalis, the fish Danio rerio, and the frog Xenopus laevis. An in vitro enzymatic assay system reveals that all DAF 36/Nvd proteins cloned so far have the ability to convert cholesterol to 7 dehydrocholesterol. Moreover, the lethality of loss of nvd function in the fruit fly Drosophila melanogaster is rescued by the expression of daf-36/nvd genes from the nematode Caenorhabditis elegans, the insect Bombyx mori, or the vertebrates D. rerio and X. laevis. These data suggest that daf-36/nvd genes are functionally orthologous across the bilaterian phylogeny. We propose that the daf-36/nvd family of proteins is a novel conserved player in cholesterol metabolism across the animal phyla. PMID- 21632548 TI - Enzyme reactivation by hydrogen peroxide in heme-based tryptophan dioxygenase. AB - An intriguing mystery about tryptophan 2,3-dioxygenase is its hydrogen peroxide triggered enzyme reactivation from the resting ferric oxidation state to the catalytically active ferrous form. In this study, we found that such an odd Fe(III) reduction by an oxidant depends on the presence of L-Trp, which ultimately serves as the reductant for the enzyme. In the peroxide reaction with tryptophan 2,3-dioxygenase, a previously unknown catalase-like activity was detected. A ferryl species (delta = 0.055 mm/s and DeltaE(Q) = 1.755 mm/s) and a protein-based free radical (g = 2.0028 and 1.72 millitesla linewidth) were characterized by Mossbauer and EPR spectroscopy, respectively. This is the first compound ES-type of ferryl intermediate from a heme-based dioxygenase characterized by EPR and Mossbauer spectroscopy. Density functional theory calculations revealed the contribution of secondary ligand sphere to the spectroscopic properties of the ferryl species. In the presence of L-Trp, the reactivation was demonstrated by enzyme assays and by various spectroscopic techniques. A Trp-Trp dimer and a monooxygenated L-Trp were both observed as the enzyme reactivation by-products by mass spectrometry. Together, these results lead to the unraveling of an over 60-year old mystery of peroxide reactivation mechanism. These results may shed light on how a metalloenzyme maintains its catalytic activity in an oxidizing environment. PMID- 21632549 TI - Imaging of intact tissue sections: moving beyond the microscope. AB - MALDI-imaging MS is a new molecular imaging technology for direct in situ analysis of thin tissue sections. Multiple analytes can be monitored simultaneously without prior knowledge of their identities and without the need for target-specific reagents such as antibodies. Imaging MS provides important insights into biological processes because the native distributions of molecules are minimally disturbed, and histological features remain intact throughout the analysis. A wide variety of molecules can be imaged, including proteins, peptides, lipids, drugs, and metabolites. Several specific examples are presented to highlight the utility of the technology. PMID- 21632550 TI - Analysis of intact protein isoforms by mass spectrometry. AB - The diverse proteome of an organism arises from such events as single nucleotide substitutions at the DNA level, different RNA processing, and dynamic enzymatic post-translational modifications. This minireview focuses on the measurement of intact proteins to describe the diversity found in proteomes. The field of biological mass spectrometry has steadily advanced, enabling improvements in the characterization of single proteins to proteins derived from cells or tissues. In this minireview, we discuss the basic technology for "top-down" intact protein analysis. Furthermore, examples of studies involved with the qualitative and quantitative analysis of full-length polypeptides are provided. PMID- 21632551 TI - Positive feedback loop between PI3K-Akt-mTORC1 signaling and the lipogenic pathway boosts Akt signaling: induction of the lipogenic pathway by a melanoma antigen. AB - The lipogenic phenotype is a metabolic hallmark of cancer cells. Sterol regulatory element-binding proteins (SREBP) are key transcriptional factors to regulate biosynthesis of cholesterol and fatty acids. It has been poorly understood how the lipogenic phenotype in cancer cells is regulated and how it augments their malignant properties. Here we describe roles of the melanoma antigen ganglioside GD3 and phosphatidylinositol 3-kinase (PI3K)-Akt-mTOR complex 1 (mTORC1) signaling in the regulation of SREBP activity, cholesterol biosynthesis, and the integrity of lipid rafts in human melanoma cells. GD3 expression induced the activation of both SREBP-1 and SREBP-2. Consequently, HMG CoA reductase expression and cholesterol biosynthesis increased. The activation of the SREBP pathway was independent of the oncogenic BRAF mutation. On the other hand, it was regulated by PI3K-Akt-mTORC1 signaling in human melanoma cells. Disruption of the signaling pathway resulted in the reduction of cholesterol in lipid rafts. Inhibition of the SREBP pathway attenuated Akt activation in lipid rafts and suppressed the growth of human melanoma cells in vitro and in vivo. These results suggest that PI3K-Akt-mTORC1 signaling is important for the integrity of lipid rafts by regulating SREBP activation and subsequent cholesterogenesis. We thus propose a positive feedback circuit in which PI3K-Akt mTORC1-SREBP signaling boosts Akt signaling in human melanoma cells expressing GD3. PMID- 21632552 TI - Prediagnostic serum levels of cytokines and other immune markers and risk of non hodgkin lymphoma. AB - Although severe immune dysregulation is an established risk factor for non Hodgkin lymphoma (NHL), it is unclear whether subclinical immune system function influences lymphomagenesis. To address this question, we conducted a nested case control study within the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial to investigate whether circulating levels of cytokines and other immune markers are associated with future risk of NHL. Selected cytokines [interleukin (IL)-4, IL-6, IL-10, and TNF-alpha] and other immune markers [soluble TNF receptor 1 (sTNF-R1), sTNF-R2, C-reactive protein, and sCD27] were measured in prediagnostic serum specimens from 297 incident NHL cases and 297 individually matched controls. ORs and 95% confidence intervals (CI) relating quartiles of analyte concentration to NHL risk were calculated by using conditional logistic regression. Statistically significant associations with increased NHL risk were observed for elevated serum levels of sTNF-R1 (quartile 4 vs. quartile 1: OR = 1.7, 95% CI: 1.1-2.8; P(trend) = 0.02) and sCD27 (OR = 5.3, 95% CI: 2.9-9.4; P(trend) < 0.0001). These associations remained in analyses of cases diagnosed longer than 6 years following blood collection (sTNF-R1: OR = 2.1, 95% CI: 1.0 4.0, P(trend) = 0.01; sCD27: OR = 4.1, 95% CI: 1.9-8.5, P(trend) = 0.0001). Elevated levels of IL-10, TNF-alpha and sTNF-R2 were also significantly associated with increased risk of NHL overall; however, these associations weakened with increasing time from blood collection to case diagnosis and were null for cases diagnosed longer than 6 years postcollection. Our findings for sTNF-R1 and sCD27, possible markers for inflammatory and B-cell stimulatory states, respectively, support a role for subclinical inflammation and chronic B cell stimulation in lymphomagenesis. PMID- 21632553 TI - Notch signaling activated by replication stress-induced expression of midkine drives epithelial-mesenchymal transition and chemoresistance in pancreatic cancer. AB - The incidence of pancreatic ductal adenocarcinoma (PDAC) nearly equals its mortality rate, partly because most PDACs are intrinsically chemoresistant and thus largely untreatable. It was found recently that chemoresistant PDAC cells overexpress the Notch-2 receptor and have undergone epithelial-mesenchymal transition (EMT). In this study, we show that these two phenotypes are interrelated by expression of Midkine (MK), a heparin-binding growth factor that is widely overexpressed in chemoresistant PDAC. Gemcitabine, the front-line chemotherapy used in PDAC treatment, induced MK expression in a dose-dependent manner, and its RNAi-mediated depletion was associated with sensitization to gemcitabine treatment. We identified an interaction between the Notch-2 receptor and MK in PDAC cells. MK-Notch-2 interaction activated Notch signaling, induced EMT, upregulated NF-kappaB, and increased chemoresistance. Taken together, our findings define an important pathway of chemoresistance in PDAC and suggest novel strategies for its clinical attack. PMID- 21632556 TI - Hybridization kinetics analysis of an oligonucleotide microarray for microRNA detection. AB - MicroRNA (miRNA) microarrays have been successfully used for profiling miRNA expression in many physiological processes such as development, differentiation, oncogenesis, and other disease processes. Detecting miRNA by miRNA microarray is actually based on nucleic acid hybridization between target molecules and their corresponding complementary probes. Due to the small size and high degree of similarity among miRNA sequences, the hybridization condition must be carefully optimized to get specific and reliable signals. Previously, we reported a microarray platform to detect miRNA expression. In this study, we evaluated the sensitivity and specificity of our microarray platform. After systematic analysis, we determined an optimized hybridization condition with high sensitivity and specificity for miRNA detection. Our results would be helpful for other hybridization-based miRNA detection methods, such as northern blot and nuclease protection assay. PMID- 21632555 TI - Hedgehog overexpression is associated with stromal interactions and predicts for poor outcome in breast cancer. AB - Hedgehog (Hh) signaling plays an important role in several malignancies but its clinical significance in breast cancer is unclear. In a cohort of 279 patients with invasive ductal carcinoma of the breast, expression of Hh ligand was significantly associated with increased risk of metastasis, breast cancer specific death, and a basal-like phenotype. A paracrine signature, encompassing high epithelial Hh ligand and high stromal Gli1, was an independent predictor for overall survival in multivariate analysis. In 2 independent histological progression series (n = 301), Hh expression increased with atypia. Hh ligand overexpression in a mouse model of basal breast cancer increased growth, induced a poorly differentiated phenotype, accelerated metastasis, and reduced survival. A stromal requirement for these effects was supported by the lack of similar Hh mediated changes in vitro, and by stromal-specific expression of Hh target genes in vivo. Furthermore, inhibition of Hh ligand with a monoclonal antibody (5E1) inhibited tumor growth and metastasis. These data suggest that epithelial-stromal Hh signaling, driven by ligand expression in carcinoma cells, promotes breast cancer growth and metastasis. Blockade of Hh signaling to peritumoral stromal cells may represent a novel therapeutic approach in some basal-like breast cancers. PMID- 21632557 TI - Alleles A and B of non-structural protein 1 of avian influenza A viruses differentially inhibit beta interferon production in human and mink lung cells. AB - Non-structural protein 1 (NS1) counteracts the production of host type I interferons (IFN-alpha/beta) for the efficient replication and pathogenicity of influenza A viruses. Here, we reveal another dimension of the NS1 protein of avian influenza A viruses in suppressing IFN-beta production in cultured cell lines. We found that allele A NS1 proteins of H6N8 and H4N6 have a strong capacity to inhibit the activation of IFN-beta production, compared with allele B from corresponding subtypes, as measured by IFN stimulatory response element (ISRE) promoter activation, IFN-beta mRNA transcription and IFN-beta protein expression. Furthermore, the ability to suppress IFN-beta promoter activation was mapped to the C-terminal effector domain (ED), while the RNA-binding domain (RBD) alone was unable to suppress IFN-beta promoter activation. Chimeric studies indicated that when the RBD of allele A was fused to the ED of allele B, it was a strong inhibitor of IFN-beta promoter activity. This shows that well-matched ED and RBD are crucial for the function of the NS1 protein and that the RBD could be one possible cause for this differential IFN-beta inhibition. Notably, mutagenesis studies indicated that the F103Y and Y103F substitutions in alleles A and B, respectively, do not influence the ISRE promoter activation. Apart from dsRNA signalling, differences were observed in the expression pattern of NS1 in transfected human and mink lung cells. This study therefore expands the versatile nature of the NS1 protein in inhibiting IFN responses at multiple levels, by demonstrating for the first time that it occurs in a manner dependent on allele type. PMID- 21632558 TI - Mutations in the G-H loop region of ephrin-B2 can enhance Nipah virus binding and infection. AB - Nipah virus (NiV) and Hendra virus (HeV) are zoonotic paramyxoviruses classified in the genus Henipavirus of the family Paramyxoviridae. The entry of henipaviruses occurs through a pH-independent membrane-fusion mechanism mediated by the cooperation of the viral attachment (G) and fusion (F) envelope glycoproteins following virion binding to susceptible host cells. Virus attachment is mediated by the interaction of the G glycoprotein with ephrin-B2 or ephrin-B3, which were identified as the functional receptors of henipavirus. Several residues of the G glycoprotein that are important for receptor binding have been determined through mutagenesis and structural analyses; however, similar approaches have not been carried out for the viral receptor ephrin-B2. Here, an alanine-scanning mutagenesis analysis was performed to identify residues of ephrin-B2 which are critical for NiV binding and entry by using an NiV-F- and G-glycoprotein pseudotyped lentivirus assay. Results indicated that the G-H loop of ephrin-B2 was indeed critical for the interaction between ephrin-B2 and NiV-G. Unexpectedly, however, some alanine-substitution mutants located in the G-H loop enhanced the infectivity of the NiV pseudotypes, in particular an L124A mutation enhanced entry >30-fold. Further analysis of the L124A ephrin-B2 mutant demonstrated that an increased binding affinity of the mutant receptor with NiV-G was responsible for the enhanced infectivity of both pseudovirus and infectious virus. In addition, cell lines that were stably expressing the L124A mutant receptor were able to support NiV infection more efficiently than the wild-type molecule, potentially providing a new target-cell platform for viral isolation or virus-entry inhibitor screening and discovery. PMID- 21632559 TI - RNA helicase retinoic acid-inducible gene I as a sensor of Hantaan virus replication. AB - Hantaan virus (HTNV) causes severe human disease. The HTNV genome consists of three ssRNA segments of negative polarity that are complexed with viral nucleocapsid (N) protein. How the human innate immune system detects HTNV is unclear. RNA helicase retinoic acid-inducible gene I (RIG-I) does not sense genomic HTNV RNA. So far it has not been analysed whether pathogen-associated molecular patterns generated during the HTNV replication trigger RIG-I-mediated innate responses. Indeed, we found that knock-down of RIG-I in A549 cells, an alveolar epithelial cell line, increases HTNV replication and prevents induction of 2',5'-oligoadenylate synthetase, an interferon-stimulated gene. Moreover, overexpression of wild-type or constitutive active RIG-I in Huh7.5 cells lacking a functional RIG-I diminished HTNV virion production. Intriguingly, reporter assays revealed that in vitro-transcribed HTNV N RNA and expression of the HTNV N ORF triggers RIG-I signalling. This effect was completely blocked by the RNA binding domain of vaccinia virus E3 protein, suggesting that dsRNA-like secondary structures of HTNV N RNA stimulate RIG-I. Finally, transfection of HTNV N RNA into A549 cells resulted in a 2 log-reduction of viral titres upon challenge with virus. Our study is the first demonstration that RIG-I mediates antiviral innate responses induced by HTNV N RNA during HTNV replication and interferes with HTNV growth. PMID- 21632560 TI - Molecular diversity and evolutionary history of rabies virus strains circulating in the Balkans. AB - Molecular studies of European classical rabies viruses (RABV) have revealed a number of geographically clustered lineages. To study the diversity of Balkan RABV, partial nucleoprotein (N) gene sequences were analysed from a unique panel of isolates (n = 210), collected from various hosts between 1972 and 2006. All of the Balkan isolates grouped within the European/Middle East Lineage, with the majority most closely related to East European strains. A number of RABV from Bosnia & Herzegovina and Montenegro, collected between 1986 and 2006, grouped with the West European strains, believed to be responsible for the rabies epizootic that spread throughout Europe in the latter half of the 20th Century. In contrast, no Serbian RABV belonged to this sublineage. However, a distinct group of Serbian fox RABV provided further evidence for the southwards wildlife mediated movement of rabies from Hungary, Romania and Serbia into Bulgaria. To determine the optimal region for evolutionary analysis, partial, full and concatenated N-gene and glycoprotein (G) gene sequences were compared. Whilst both the divergence times and evolutionary rates were similar irrespective of genomic region, the 95 % highest probability density (HPD) limits were significantly reduced for full N-gene and concatenated NG-gene sequences compared with partial gene sequences. Bayesian coalescent analysis estimated the date of the most common recent ancestor of the Balkan RABV to be 1885 (95 % HPD, 1852 1913), and skyline plots suggested an expansion of the local viral population in 1980-1990, which coincides with the observed emergence of fox rabies in the region. PMID- 21632561 TI - Herpes simplex virus type 2 virion host shutoff protein suppresses innate dsRNA antiviral pathways in human vaginal epithelial cells. AB - Viruses that establish persistent infections have evolved numerous strategies to evade host innate antiviral responses. We functionally assessed the role of herpes simplex virus type 2 (HSV-2) virion host shutoff (vhs) protein on innate immune sensing pathways in human vaginal epithelial cells (VK2 ECs). Infection of cells with wild-type (WT) HSV-2 significantly decreased expression of innate immune sensors of viral infection, Toll-like receptor (TLR)2, TLR3, retinoic acid inducible gene I (RIG-I) and melanoma differentiation-associated gene 5 (Mda-5), relative to cells infected with a mutant that lacks vhs (vhsB) or mock-infected cells. Transfection with HSV-2 vhs similarly decreased expression of TLR2, TLR3, RIG-I and Mda-5, which was also confirmed in human embryonic kidney (HEK) 293 cells. vhsB infection of VK2 cells caused robust increases in the active form of interferon regulatory factor (IRF)3 and its translocation to the nucleus compared with the WT. Additionally, IRF3 activation by Sendai virus and polyinosinic : polycytidylic acid-induced stimulation of beta interferon (IFN-beta) was significantly inhibited in vhs-transfected cells. Overall, our findings provide the first evidence that HSV-2 vhs plays roles in selectively inhibiting TLR3 and RIG-I/Mda-5, as well as TLR2-mediated antiviral pathways for sensing dsRNA and effectively suppresses IFN-beta antiviral responses in human vaginal ECs. PMID- 21632562 TI - A novel mechanism for the inhibition of interferon regulatory factor-3-dependent gene expression by human respiratory syncytial virus NS1 protein. AB - Human respiratory syncytial virus (RSV), a leading cause of respiratory tract infections in infants, inhibits type I interferon (IFN)-dependent signalling, as well as IFN synthesis. RSV non-structural protein NS1 plays a significant role in this inhibition; however, the mechanism(s) responsible is not fully known. The transcription factor interferon regulatory factor (IRF)-3 is essential for viral induced IFN-beta synthesis. In this study, we found that NS1 protein inhibits IRF 3-dependent gene transcription in constitutively active IRF-3 overexpressing cells, demonstrating that NS1 directly targets IRF-3. Our data also demonstrate that NS1 associates with IRF-3 and its transcriptional coactivator CBP, leading to disrupted association of IRF-3 to CBP and subsequent reduced binding of IRF-3 to the IFN-beta promoter without blocking viral-induced IRF-3 phosphorylation, nuclear translocation and dimerization, thereby identifying a novel molecular mechanism by which RSV inhibits IFN-beta synthesis. PMID- 21632563 TI - Genetic diversification of H5N1 highly pathogenic avian influenza A virus during replication in wild ducks. AB - Highly pathogenic avian influenza A virus subtype H5N1 can potentially generate novel variants during replication of infected hosts. To determine which H5N1 variants predominate in wild birds, we determined the sequences of RT-PCR amplified viral genes from several organs of infected chickens and ducks from Egypt, where H5N1 outbreaks in birds are endemic. Comparison of the sequences in viruses from trachea, lung, brain and liver revealed diversification with different amino acid substitutions in different ducks, but no diversification in chickens. These specific amino acid substitutions were rare among viruses currently circulating in Egypt. In addition, the H5N1 variants showed distinct growth kinetics in duck, canine and human cells. Our findings suggested that ducks can generate H5N1 variants with novel amino acid substitutions that might serve as aetiological agents for new influenza virus outbreaks and epidemics. PMID- 21632564 TI - Lack of type-specific concordance between human papillomavirus (HPV) serology and HPV DNA detection in the uterine cervix and oral mucosa. AB - There is limited knowledge about longitudinal genotype-specific concordance between human papillomavirus (HPV) serology and co-existent presence of HPV DNA in the uterine cervix. The role of oral HPV infections in inducing serological response is unclear, as is the effect of HPV antibodies on the outcome of oral HPV infections. The present study is part of the Finnish Family HPV Study designed to evaluate dynamics of HPV infections within families. Here, we correlated the point prevalence of HPV6, 11, 16, 18 and 45 antibodies and concomitant genotype-specific HPV DNA detection in cervical and oral samples of 323 mothers during their 3 year (mean 37.5 months) follow-up. The mean age of these pregnant mothers at enrolment (third trimester) was 25.5 years. HPV antibodies were analysed with multiplex HPV serology and HPV genotyping was performed using a Multimetrix kit (Progen Biotechnik). There was no concordance between cervical DNA detection and co-existent seropositivity, and the same was true even in samples taken 12 months apart. Women who cleared their cervical HPV16 infection had the highest HPV16 antibody levels, whereas those who acquired incident HPV16 infections had the lowest antibody levels. Neither the presence nor the dynamics of oral HPV DNA had any correlation with HPV serology. PMID- 21632565 TI - Differential effect of acute and persistent Junin virus infections on the nucleo cytoplasmic trafficking and expression of heterogeneous nuclear ribonucleoproteins type A and B. AB - Heterogeneous nuclear ribonucleoproteins A and B (hnRNPs A/B), cellular RNA binding proteins that participate in splicing, trafficking, translation and turnover of mRNAs, have been implicated in the life cycles of several cytoplasmic RNA viruses. Here, we demonstrate that silencing of hnRNPs A1 and A2 significantly reduces the replication of the arenavirus Junin virus (JUNV), the aetiological agent of Argentine haemorrhagic fever. While acute JUNV infection did not modify total levels of expression of hnRNPs A/B in comparison with uninfected cells, non-cytopathic persistent infection exhibited low levels of these cell proteins. Furthermore, acutely infected cells showed a cytoplasmic relocalization of overexpressed hnRNP A1, probably related to the involvement of this protein in virus replicative cycle. This cytoplasmic accumulation was also observed in cells expressing viral nucleoprotein (N), and co-immunoprecipitation studies revealed the interaction between hnRNP A1 and N protein. By contrast, a predominantly nuclear distribution of overexpressed hnRNP A1 was found during persistent infection, even in the presence of endogenous or overexpressed N protein, indicating a differential modulation of nucleo-cytoplasmic trafficking in acute and persistent JUNV infections. PMID- 21632566 TI - Co-existence of multiple strains of two novel porcine bocaviruses in the same pig, a previously undescribed phenomenon in members of the family Parvoviridae, and evidence for inter- and intra-host genetic diversity and recombination. AB - Despite the recent discovery of novel bocaviruses from porcine samples, their genetic evolution and diversity are poorly understood. This study reports the identification and complete genome characterization of two novel parvoviruses, porcine bocavirus 3 (PBoV3) and porcine bocavirus 4 (PBoV4), from various porcine tissues/samples, displaying marked intra- and inter-host genetic diversity, with recombination events. Bocaviruses were detected by PCR among 16.5 % (55/333) of porcine samples (lymph nodes, serum, nasopharyngeal and faecal samples) from healthy, sick or deceased pigs from farms and a slaughterhouse in Hong Kong. As marked nucleotide polymorphisms were observed in the partial VP1 sequences, complete VP1 genes from one nasopharyngeal and three faecal specimens were cloned and sequenced, which suggested the presence of two different bocaviruses and demonstrated significant intra- and inter-host genetic diversity. Complete genome sequences revealed the presence of two bocaviruses, PBoV3 and PBoV4, in a faecal and nasopharyngeal specimen, respectively, with two genotypes, PBoV4-1 and PBoV4 2, in the latter. Their genomes encoded three ORFs, characteristic of bocaviruses. Phylogenetic analysis showed that they were distantly related to other bocaviruses, forming a distinct cluster within the genus. Recombination analysis showed possible recombination events among VP1 sequences of PBoV4 strains from a faecal specimen, with two breakpoints identified (with a 68 and 71 bp region), suggesting that different strains/variants within the same host could have arisen from recombination. This is the first report describing marked sequence diversity and the co-existence of two viruses of the family Parvoviridae within the same host, which may have originated from and, in turn, facilitated recombination. PMID- 21632567 TI - Antigen-presenting cells of haematopoietic origin prime cytomegalovirus-specific CD8 T-cells but are not sufficient for driving memory inflation during viral latency. AB - Expansion of the CD8 T-cell memory pool, also known as 'memory inflation', for certain but not all viral epitopes in latently infected host tissues is a special feature of the immune response to cytomegalovirus. The L(d)-presented murine cytomegalovirus (mCMV) immediate-early (IE) 1 peptide is the prototype of an epitope that is associated with memory inflation. Based on the detection of IE1 transcripts in latently infected lungs it was previously proposed that episodes of viral gene expression and antigenic activity due to desilencing of a limited number of viral genes may drive epitope-specific memory inflation. This would imply direct antigen presentation through latently infected host tissue cells rather than cell death-associated cross-presentation of viral antigens derived from productively infected cells through uninfected, professional antigen presenting cells (profAPCs). To address the role of bone marrow-derived profAPCs in CD8 T-cell priming and memory to mCMV, we have used here a combined sex mismatched and MHC class-I mismatched dual-marker bone marrow chimera model in which presentation of the IE1 epitope is restricted to donor-derived sry(+)L(d+) cells of haematopoietic differentiation lineages. Successful CD8 T-cell priming specific for the L(d)- and D(d)-presented inflationary epitopes IE1 and m164, respectively, but selective failure in IE1 epitope-specific memory inflation in these chimeras indicates different modes of antigen presentation involved in CD8 T-cell priming and memory inflation. These data suggest that memory inflation during mCMV latency requires expression of the epitope-presenting MHC class-I molecule by latently infected non-haematopoietic host tissue cells and thus predicts a role for direct antigen presentation in memory inflation. PMID- 21632568 TI - H2B homology region of major immediate-early protein 1 is essential for murine cytomegalovirus to disrupt nuclear domain 10, but is not important for viral replication in cell culture. AB - Cytomegalovirus (CMV) major immediate-early protein 1 (IE1) has multiple functions and is important for efficient viral infection. As does its counterpart in human CMV, murine CMV (MCMV) IE1 also functions as a disruptor of mouse-cell nuclear domain 10 (ND10), where many different gene-regulation proteins congregate. It still remains unclear how MCMV IE1 disperses ND10 and whether this dispersion could have any effect on viral replication. MCMV IE1 is 595 aa long and has multiple functional domains that have not yet been fully analysed. In this study, we dissected the IE1 molecule by truncation and/or deletion and found that the H2B homology domain (amino acid sequence NDIFERI) is required for the dispersion of ND10 by IE1. Furthermore, we made additional deletions and point mutations and found that the minimal truncation in the H2B homology domain required for IE1 to lose the ability to disperse ND10 is just 3 aa (IFE). Surprisingly, the mutated IE1 still interacted with PML and co-localized with ND10 but failed to disperse ND10. This suggests that binding to ND10 key protein is essential to, but not sufficient for, the dispersal of ND10, and that some other unknown mechanism must be involved in this biological procedure. Finally, we generated MCMV with IFE-deleted IE1 (MCMVdlIFE) and its revertant (MCMVIFERQ). Although MCMVdlIFE lost the ability to disperse ND10, plaque assays and viral gene production assays showed that the deletion of IFE did not increase viral replication in cell culture. We conclude that the dispersion of ND10 appears not to be important for MCMV replication in a mouse-cell culture. PMID- 21632569 TI - Defective interfering influenza A virus protects in vivo against disease caused by a heterologous influenza B virus. AB - Influenza A and B viruses are major human respiratory pathogens that contribute to the burden of seasonal influenza. They are both members of the family Orthomyxoviridae but do not interact genetically and are classified in different genera. Defective interfering (DI) influenza viruses have a major deletion of one or more of their eight genome segments, which renders them both non-infectious and able to interfere in cell culture with the production of infectious progeny by a genetically compatible, homologous virus. It has been shown previously that intranasal administration of a cloned DI influenza A virus, 244/PR8, protects mice from various homologous influenza A virus subtypes and that it also protects mice from respiratory disease caused by a heterologous virus belonging to the family Paramyxoviridae. The mechanisms of action in vivo differ, with homologous and heterologous protection being mediated by probable genome competition and type I interferon (IFN), respectively. In the current study, it was shown that 244/PR8 also protects against disease caused by a heterologous influenza B virus (B/Lee/40). Protection from B/Lee/40 challenge was partially eliminated in mice that did not express a functional type I IFN receptor, suggesting that innate immunity, and type I IFN in particular, are important in mediating protection against this virus. It was concluded that 244/PR8 has the ability to protect in vivo against heterologous IFN-sensitive respiratory viruses, in addition to homologous influenza A viruses, and that it acts by fundamentally different mechanisms. PMID- 21632570 TI - War injuries. PMID- 21632571 TI - Prevalence and social determinants of suicidal behaviours among college youth in India. AB - BACKGROUND: Little is known about the prevalence of and factors associated with suicide ideation and suicide attempts among college youth in India. AIMS: The aims were to estimate the prevalence of suicidal behaviours among Indian college students and identify potential risk factors for suicide ideation and attempts. METHOD: The study surveyed 1,817 undergraduate college students aged 18-24 years in Ahmedabad, Gujarat, with a questionnaire that assessed suicidal behaviours as well as stressful situations and life events. Logistic regression analysis was used to assess risk factors. RESULTS: The prevalence of lifetime suicide ideation and lifetime suicide attempts was 11.7% and 4.0%, respectively. Suicide ideation was predicted by female gender, odds ratio (OR) = 1.41, 95% CI 1.01, 1.97, economic stress, OR = 1.17, 95% CI 1.11, 1.24, stress due to life events involving religious violence in the community, OR = 1.43, 95% CI 1.15, 1.78, and life events involving caste conflicts or caste discrimination, OR = 1.28, 95% CI 1.13, 1.46. Female gender and caste-related life events were not significantly associated with suicide attempts but economic stress and stressful experience of religious conflict continued to be significantly associated with lifetime suicide attempt, OR = 1.19, 95% CI 1.08, 1.31, and OR = 1.58, 95% CI 1.14, 2.17, respectively. CONCLUSIONS: In this sample, college students from low socioeconomic classes who faced economic difficulties, and students who experienced distress as a result of caste discrimination or caste conflict, and communal unrest, were at a higher risk for suicidal behaviour. PMID- 21632572 TI - Effects of vitamin E on reproductive hormones and testis structure in chronic dioxin-treated mice. AB - The purpose of this study was to investigate the effects of vitamin E on reproductive hormones and testis structure in mice treated with 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD). Five experimental groups of a combination of TCDD and vitamin E were designed as follows: 0 ng/kg/d and 0 mg/kg/d (control group), 100 ng/kg/d and 0 mg/kg/d (Group I), 100 ng/kg/d and 20 mg/kg/d (Group II), 100 ng/kg/d and 100 mg/kg/d (Group III), and 100 ng/kg/d and 500 mg/kg/d (Group IV) respectively. Vitamin E and TCDD were given by oral gavage for 7 weeks. The results demonstrated that TCDD decreased the levels of brain gonadotropin releasing hormone (GnRH), testis luteinizing hormone (LH) and follicle stimulating hormone (FSH), serum testosterone and testis spermatozoa number, and damaged testis structure. Vitamin E at 20 mg/kg alleviated the decrease of GnRH; vitamin E at 20, 100, and 500 mg/kg antagonized the decline of LH and FSH; vitamin E at 20 and 100 mg/kg reversed the decrease of testosterone and spermatozoa number; and vitamin E at 100 mg/kg decreased the damage of the testis structure caused by TCDD. The results indicate that vitamin E antagonizes the reproductive endocrine toxicity and alleviates the changes in testicular structure caused by TCDD. PMID- 21632573 TI - The evaluation of the genotoxic and oxidative damage potentials of Ulothrix tenuissima (Kutz.) in vitro. AB - Several alga species are known to produce a variety of toxic metabolites that pose a threat to aquatic organisms, animals and humans. Moreover, these metabolites have been thought to cause serious diseases including certain cancers and neurodegenerative disorders. On the other hand, Ulothrix is a genus of filamentous green algae, generally found in fresh water and marine and abundantly available in some lakes and rivers of Turkey. To our best knowledge, no study has been performed to assess the genotoxic and biochemical effects of U. tenuissima on cultured human blood cells. Therefore, in order to determine clastogenic or aneugenic effects of aqueous alga extracts the micronucleus assay was carried out. Nuclear division index (NDI) in peripheral lymphocytes was also analyzed for cytotoxicity evaluations. In addition, biochemical parameters (total antioxidant capacity (TAC) and total oxidative stress (TOS)) were examined to determine oxidative effects. For this aim, we obtained heparinized blood samples from three healthy persons. The alga samples were collected from Porsuk Pond in Hasankale (Erzurum, Turkey) in summer period of the year 2010. The aqueous extracts of this species were added to cultures at different concentrations (0 to 5000 ppm) for 72 h. Our results showed that this alga did not cause any statistically important changes in the rates of studied genotoxicity endpoint. But dose-dependent alterations were observed in TAC and TOS levels and NDI rates. In conclusion, U. tenuissima was found to be non-genotoxic but caused sterility at higher concentrations due to oxidative stress. PMID- 21632574 TI - Electroencephalogram, cognitive state, psychological disorders, clinical symptom, and oxidative stress in horticulture farmers exposed to organophosphate pesticides. AB - The aim of this paper was to study the toxicity of organophosphate (OP) pesticides in exposed farmers for electroencephalography, cognitive state, psychological disorders, clinical symptom, oxidative stress, acetylcholinesterase, and DNA damage. A comparative cross-sectional analysis was carried out in 40 horticulture farmers who were exposed to OPs in comparison to a control group containing 40 healthy subjects with the same age and sex and education level. Lipid peroxidation (LPO), superoxide dismutase (SOD), catalase, glutathione peroxidase, DNA damage, total antioxidant capacity (TAC), total thiol molecules, and acetylcholinesterase (AChE) activity were measured in the blood of subjects. Clinical examination and complete blood test were undertaken in order to record any abnormal sign or symptoms. Cognitive function, psychological symptoms, and psychological distress were examined and recorded. Comparing with controls, the farmers showed higher blood levels of SOD and LPO while their TAC decreased. Farmers showed clinical symptoms such as eczema, breathing muscle weakness, nausea, and saliva secretion. Regarding cognitive function, the orientation, registration, attention and calculation, recall, and language were not significantly different in farmers and controls. Among examinations for psychological distress, only labeled somatization was significantly higher in farmers. The present findings indicate that oxidative stress and inhibition of AChE can be seen in chronically OP-exposed people but incidence of neuropsychological disorders seems a complex multivariate phenomenon that might be seen in long-term high-dose exposure situations. Use of supplementary antioxidants would be useful in the treatment of farmers. PMID- 21632575 TI - Anti-apoptotic effects of curcumin on cadmium-induced apoptosis in rat testes. AB - Cadmium (Cd) is one of the environmental pollutants affecting various tissues and organs including testis. The aim of this study was to investigate the anti apoptotic effects of curcumin (Cur) on Cd-induced apoptosis in rat testes. The rats were randomly allotted into one of three experimental groups: control, Cd treated and Cd treated with Cur; each group contained 10 animals. The control group received 2 ml/day of dimethyl sulfoxide (DMSO). To induce toxicity, Cd (1 mg/kg body weight) was dissolved in normal saline and subcutaneously injected into rats for 4 weeks. The rats in Cur-treated group was given a daily dose of 100 mg/kg of Cur for 4 weeks. To date, no examinations of the anti-apoptotic properties of Cur on Cd-induced apoptosis in rat testes have been reported. The mean seminiferous tubule diameter, mean testicular biopsy score values and serum testosterone levels were significantly decreased in Cd-treated groups were compared to the control group. Furthermore, the Cur-treated animals showed an improved histological appearance and serum testosterone levels in Cd-treated group. Our data indicate a significant reduction in the activity of in situ identification of apoptosis using terminal dUTP nick end-labeling in testis tissues of the Cd-treated group with Cur therapy. The present study showed that Cur treatment protected testes against toxic effects of Cd. We believe that further preclinical research into the utility of Cur may indicate its usefulness as a potential treatment on the spermatogenesis after testicular injury caused by Cd-treated rats. PMID- 21632576 TI - Nutritional evaluation of alcoholic inpatients admitted for alcohol detoxification. AB - AIMS: To assess nutritional risk of alcoholic patients admitted for alcohol detoxification. METHODS: Screening of nutritional risk of alcoholic patients using the Malnutrition Universal Screening Tool. RESULTS: Fifty-three percentage patients at presentation were rated as being at medium or high risk of malnutrition. CONCLUSION: Malnutrition should be actively considered and screened for in alcoholic patients admitted for alcohol detoxification due to its high prevalence and benefits obtained from treatment. PMID- 21632577 TI - Reutericyclin and related analogues kill stationary phase Clostridium difficile at achievable colonic concentrations. AB - OBJECTIVES: The stationary phase of Clostridium difficile, which is associated with the symptoms of the diarrhoeal disease, is refractory to antibiotic killing. The aim of this study was to explore whether probiotic-derived reutericyclin and related synthetic analogues could kill stationary phase C. difficile at concentrations achievable in the gastrointestinal tract. METHODS: The bactericidal activities of reutericyclin and lead compound derivatives were examined against logarithmic and stationary phase cultures of different C. difficile strains. The absorption of compounds across the intestinal epithelia was tested using the Caco-2 permeability model. RESULTS: Unlike vancomycin and metronidazole, reutericyclins demonstrated concentration-dependent killing, being rapidly bactericidal against both logarithmic and stationary phase cells, at low concentrations (0.09-2 mg/L). The intestinal absorption of unmodified reutericyclin was poor and comparable to that of vancomycin. However, this property varied significantly for the synthetic reutericyclin analogues, ranging from well absorbed to non-absorbed. The non-absorbable compounds were highly effluxed, suggesting this parameter could be modulated to obtain agents with superior efficacy. CONCLUSIONS: Reutericyclins showed excellent potency against the lethal non-growing stage of C. difficile at concentrations that may be attained in the gastrointestinal tract. Since these agents represent novel potential treatments for C. difficile infection, further development of this compound class is warranted. PMID- 21632578 TI - Ertapenem in plasma and peritoneal fluid from patients with severe intra abdominal infections. PMID- 21632579 TI - Empirical modified sequential therapy containing levofloxacin and high-dose esomeprazole in second-line therapy for Helicobacter pylori infection: a multicentre clinical trial. AB - OBJECTIVES: Sequential therapy appears to achieve a higher Helicobacter pylori eradication rate than triple therapy. We assessed the efficacy and tolerability of modified sequential therapy containing levofloxacin and high-dose esomeprazole in second-line therapy. METHODS: Patients who failed first-line triple therapy with clarithromycin, amoxicillin and a proton pump inhibitor were eligible in this multicentre trial. Eligible patients were treated with esomeprazole 40 mg and amoxicillin 1 g for the first 5 days, followed by esomeprazole 40 mg, levofloxacin 250 mg and metronidazole 500 mg for another 5 days (all given twice daily). Eradication was confirmed with a (13)C-urea breath test 6 weeks after therapy. Drug susceptibility, presence/absence of gyrA mutation and CYP2C19 genotype were also determined. RESULTS: A total of 142 patients were enrolled. The eradication rate was 95.1% [135/142, 95% confidence interval (CI) 91.5% 98.6%] in the intention-to-treat analysis and 96.4% (133/138, 95% CI 93.3%-99.5%) in the per protocol analysis. Four patients (2.8%) failed to take at least 80% of the drugs due to adverse effects. The eradication rates were 50% (4/8) and 97.7% (43/44) in patients with and without metronidazole resistance, respectively (P = 0.001). The eradication rates were 84.6% (11/13) and 95.1% (58/61) in patients with and without gyrA mutation, respectively (P = 0.210). The eradication rates were not affected by the CYP2C19 polymorphism (P = 0.421). CONCLUSIONS: This modified sequential therapy achieved an excellent eradication rate (>95%) in second-line treatment and the eradication rate appeared to be affected by metronidazole resistance. PMID- 21632580 TI - Myocarditis as a precipitating factor for heart failure: evaluation and 1-year follow-up using cardiovascular magnetic resonance and endomyocardial biopsy. AB - AIMS: The aim of this study was to evaluate myocarditis as a precipitating factor for heart failure using cardiovascular magnetic resonance (CMR) and endomyocardial biopsy. METHODS AND RESULTS: Eighty-five patients with suspected myocarditis and 20 controls were evaluated. Seventy-one patients with positive CMR were referred for endomyocardial biopsy and re-evaluation after 1 year. Cardiovascular magnetic resonance was performed using STIR T2-weighted (T2W), early T1-weighted (EGE), and late gadolinium-enhanced (LGE) images. Immunohistological and polymerase chain reaction (PCR) analysis of myocardial specimens was employed. In patients with myocarditis, T2 and EGE were increased compared with controls (2.6 +/- 0.9 vs. 1.57 +/- 0.13, P < 0.001 and 7.9 +/- 5.5 vs. 3.59 +/- 0.08, P < 0.001, respectively). Late gadolinium enhancement was found in all myocarditis patients. Endomyocardial biopsy performed in 50 of 71 patients with positive CMR showed positive immunohistology in 48% and presence of infectious genomes in 80% (mainly Chlamydia, Herpes, and Parvovirus B19). Left ventricular ejection fraction (LVEF) was significantly decreased compared with controls (47.7 +/- 19.2 vs. 64 +/- 0.2, P < 0.001). After 1 year, CMR showed normalization of T2 and EGE, and decreased LGE. Left ventricular ejection fraction increased in 36.5% of patients, remained stable in 56.5% and decreased in 7% of patients, in whom biopsy showed persistence of the initial infective agents. A negative correlation was identified between EGE, LGE, and LVEF. Patients with positive biopsies had lower LVEFs. CONCLUSION: In a Greek population with myocarditis, Chlamydia with viruses was a common finding. Cardiovascular magnetic resonance and PCR proved useful for the detection of myocarditis; EGE and LGE had the best correlation for the development of heart failure. Persistence of the initially detected infective agents was identified in patients who deteriorated further. PMID- 21632581 TI - Usefulness of the lowest VE/VCO2 ratio measured during exercise in heart failure. PMID- 21632582 TI - Measuring cosmic-ray exposure in aircraft using real-time personal dosemeters. AB - Aircrew exposure to radiation was measured on several long-haul flights using two small commercial electronic personal dosemeters: one was a photon dosemeter, the NRF20; the other was a neutron dosemeter, the NRY21-both manufactured by Fuji Electric Systems Co. Ltd. for radiation protection at nuclear facilities. Non neutron doses were measured using the photon dosemeter, and neutron doses were measured using the neutron dosemeter. The measured non-neutron doses at commercial aviation altitudes agree with the EPCARD (European Program Package for the Calculation of Aviation Route Doses) dose calculation within a difference of 8 %. However, the recorded neutron doses were 5-15 times larger than the EPCARD calculation. These over-measurements are dependent on cut-off rigidities. PMID- 21632583 TI - Radioactivity of coals and ashes from Catalagzi coal-fired power plant in Turkey. AB - The Catalagzi coal-fired power plant (CFPP) is the Turkish CFPP that uses the hard coals produced in Zonguldak, located in the West Black Sea region of the country. Gamma-ray spectrometry was used to determine (226)Ra, (232)Th and (40)K contents in pulverised coal, bottom ash and fly ash samples. The natural radionuclide concentrations in pulverised coal ranged from 29 to 61 Bq kg(-1) for (226)Ra, from 32 to 55 Bq kg(-1) for (232)Th and from 229 to 414 Bq kg(-1) for (40)K. The fly ash fraction gave concentrations ranging from 80 to 98 Bq kg(-1) for (226)Ra, from 64 to 85 Bq kg(-1) for Th and from 754 to 992 Bq kg(-1) for (40)K, respectively. The enrichment factors from coal to fly ashes are 1.7, 2.24 and 2.6 for (232)Th, (226)Ra and (40)K, respectively. Therefore, it is advisable to monitor the environmental impact of the power plant. PMID- 21632584 TI - Time dependence of 50 Hz magnetic fields in apartment buildings with indoor transformer stations. AB - Twenty-four hour measurements of 50 Hz magnetic fields (MFs) in apartment buildings containing transformer stations have been performed. The apartments were classified into four types, according to their location relative to the transformer room. Temporal correlation coefficients between the MF in various apartments, as well as between MF and transformer load curves, were calculated. It was found that, in addition to their high average MF, the apartments located right above the transformer room also exhibit unique temporal correlation properties. PMID- 21632585 TI - Radon prevention in new construction in Finland: a nationwide sample survey in 2009. AB - The building code for radon prevention and the associated practical guidelines were revised in Finland in 2003-2004. Thereafter, preventive measures have become more common and effective and indoor radon concentrations have been markedly reduced. In this study, the indoor radon concentration was measured in 1500 new low-rise residential houses. The houses were randomly selected and represented 7 % of the houses that received building permission in 2006. The average radon concentration of all the houses measured, which were completed in 2006-2008, was 95 Bq m(-3), the median being 58 Bq m(-3). The average was 33 % lower than in houses completed in 2000-2005. The decrease was 47 % in provinces with the highest indoor radon concentration and 26 % elsewhere in the country. In houses with a slab-on-ground foundation that had both passive radon piping and sealing measures carried out using a strip of bitumen felt in the joint between the foundation wall and floor slab, the radon concentration was on average reduced by 57 % compared with houses with no preventive measures. Preventive measures were taken nationwide in 54 % of detached houses and in provinces with the highest radon concentration in 92 % of houses. PMID- 21632586 TI - Present and future studies on environmental radioactivity in Japan. AB - In this paper, recent studies on environmental radioactivity are briefly reviewed. Considering the growing dependency on nuclear energy in Asia, emphasis has been given to (14)C cycle at the ground surface including rice paddy fields and to long-range atmospheric transport of natural and anthropogenic radioactive materials. Studies on the dynamic behaviour of (14)C are being carried out and some important experimental results are becoming available for models of (14)C migration in rice paddy. Long-range atmospheric transport models have been improved substantially by using sophisticated physical models and simulation techniques to improve emergency preparedness. Although our knowledge on radon flux distribution in Asia has been improved and radon has become a more reliable atmospheric tracer, we still need more intensive measurements of exhalation flux and atmospheric concentration of radon and radium content in soil. PMID- 21632587 TI - Determination of thoron equilibrium factor from simultaneous long-term thoron and its progeny measurements. AB - With 3-month simultaneous measurements of thoron and its progeny concentrations in the lowest floors of 109 homes, the geometric mean (GM) of thoron equilibrium factor was determined to be 0.019 with a geometric standard deviation (GSD) of 3.63. Combined with the analysis from results obtained from a previous study in 138 homes, the GM of thoron equilibrium factor was determined to be 0.022 with a GSD of 3.02. The results indicate that the F value of 0.02 for thoron recommended by UNSCEAR is a reasonable value for those Canadian homes tested. PMID- 21632588 TI - Therapeutic hypothermia for management of neonatal asphyxia: what nurses need to know. AB - Birth asphyxia can induce a cascade of reactions that result in altered brain function known as hypoxic-ischemic encephalopathy. Possible outcomes for survivors of birth asphyxia vary widely, from a normal outcome to death, with a wide range of disabilities in between, including long-term neurodevelopmental disability, cerebral palsy, neuromotor delay, and developmental delay. Treatment of hypoxic-ischemic encephalopathy has centered on dampening or blocking the biochemical pathways that lead to death of neuronal cells. The reduction of body temperature by 3oC to 5oC less than normal body temperature can reduce cerebral injury. At Mount Sinai Hospital in Toronto, Ontario, the goal of therapeutic hypothermia is to achieve a rectal temperature of 33oC to 34oC, and the protocol is started within 6 hours after birth. The hypothermia is maintained for 72 hours, and then the infant is gradually warmed to normal body temperature (36.8oC 37oC). The protocol and nursing implications are presented. PMID- 21632590 TI - Survival from acute myocardial infarction: does your hospital have the attributes that matter most? PMID- 21632591 TI - Nonpharmacological interventions to manage common symptoms in patients receiving mechanical ventilation. AB - Patients receiving mechanical ventilation can experience symptoms such as pain, anxiety, agitation, and lack of sleep while in the intensive care unit, all of which can affect healing. Nonpharmacological complementary therapies can be used as adjuncts to sedatives and analgesics. By incorporating appropriate use of complementary therapies in conjunction with mainstream medical therapies, nurses can decrease patients' anxiety, promote sleep, and promote a healing environment to improve outcomes. Minimizing noise and providing access to natural light help promote a healing environment. Methods to promote sleep include relaxation techniques such as progressive muscle relaxation and massage and communication with patients' and their families to determine the patients' normal sleep patterns. Complementary therapies to relieve anxiety and agitation include music intervention, imagery, presence, and animal-assisted therapy. PMID- 21632592 TI - Overview of mechanical ventilatory support and management of patient- and ventilator-related responses. AB - Nurses must be knowledgeable about the function and limitations of ventilator modes, causes of respiratory distress and dyssynchrony with the ventilator, and appropriate management in order to provide high-quality patient-centered care. Prompt recognition of problems and action by the nurse may resolve acute respiratory distress, dyspnea, and increased work of breathing and prevent adverse events. This article presents an overview of mechanical ventilation modes and the assessment and management of dyspnea and patient-ventilator dyssynchrony. Strategies to manage patients' responses to mechanical ventilatory support and recommendations for staff education also are presented. PMID- 21632593 TI - Disposable vs reusable electrocardiography leads in development of and cross contamination by resistant bacteria. AB - Hospital-acquired infections caused by antibacterial-resistant microorganisms are associated with high mortality and morbidity rates and markedly affect hospital economics. The expense became greater in 2008 when reimbursement for treatment of hospital-acquired infections was no longer provided by Medicare. Infections caused by cross-contamination with resistant bacteria can be eliminated by 3 methods: kill the bacteria before resistance develops, stop bacteria from communicating and acquiring resistance, and eliminate the pathway from one patient to another. Because electrocardiography wires cannot be completely disinfected 100% of the time, they may be contributing to the growth of resistant bacteria. The many pathways provided by reusable wires for cross-contamination with resistant bacteria increase the risk for hospital-acquired infection when these wires are used. Disposable electrocardiography leads eliminate risk of infection through these pathways. Adoption of disposable electrocardiography leads as an adjunct to an overall infection control program can decrease infection rates in acute health care facilities. PMID- 21632594 TI - Flexible bronchoscopy assisted by noninvasive positive pressure ventilation. AB - Noninvasive positive pressure ventilation is an alternative to intubation in critically ill patients with respiratory insufficiency or poor gas exchange who may require flexible bronchoscopy for diagnostic or therapeutic purposes. This ventilatory technique might help decrease the risk of bronchoscopy-related complications in patients with refractory hypoxemia, postoperative respiratory distress, severe emphysema, obstructive sleep apnea, and obesity hypoventilation syndrome and allows bronchoscopic assessment of patients with severe dyspnea from expiratory central airway collapse. In this review, the physiological rationale, indications, contraindications, techniques, and monitoring requirements for flexible bronchoscopy assisted by noninvasive positive pressure ventilation are described, with an emphasis on the role of critical care nurses in this procedure. PMID- 21632595 TI - Progressive care units: different but the same. AB - Progressive care units are increasingly common in hospitals in the United States. These units are often used to bridge the gap between intensive care units and medical-surgical units, with the goal of providing cost-effective, high-quality, safe care. Although progressive care units today may seem to provide care for a wide variety of patients, these patients do share certain typical features: they require a high intensity of nursing care and/or a high level of surveillance. Nurses working in progressive care units all must have certain basic competencies. Those core competencies have been identified and should form the basis for education and training of progressive care nurses. PMID- 21632596 TI - Documentation of others' work in the electronic health record. PMID- 21632597 TI - Effects of a spirituality training program for nurses on patients in a progressive care unit. PMID- 21632600 TI - Childhood appendectomy, tonsillectomy, and risk for premature acute myocardial infarction--a nationwide population-based cohort study. AB - AIMS: Although inflammation contributes to cardiovascular disease, the associations of appendectomy and tonsillectomy, which remove mucosa-associated lymphoid tissue, with risk of acute myocardial infarction (AMI) are unknown. Our aim was to assess the association between these operations performed in childhood and AMI risk later in life. METHODS AND RESULTS: We conducted a prospective matched cohort study among all Swedish residents born between 1955 and 1970. A national register identified all appendectomies and tonsillectomies. For each patient undergoing appendectomy or tonsillectomy, we randomly selected five controls without the history of the respective operation, matched on sex, age, and county of residence. Participants were followed for fatal and non-fatal AMI for an average of 23.5 years. Because appendiceal and tonsillar tissues have reduced function after adolescence, our primary analyses were restricted to individuals below age 20 at the time of surgery (54 449 appendectomies and 27 284 tonsillectomies). We derived hazard ratios (HRs) from proportional hazard models adjusted for parental occupation and parental history of AMI. Operations before 20 years of age were associated with an increased risk for AMI (417 and 216 events in the appendectomy and tonsillectomy datasets, respectively), with adjusted HRs of 1.33 [95% confidence interval (CI), 1.05-1.70] for appendectomy and 1.44 (95% CI, 1.04-2.01) for tonsillectomy. This association was graded, with the highest risk among those undergoing both procedures, and generally similar among both males and females. Appendectomy and tonsillectomy performed at or above 20 years of age were not associated with the risk of AMI. CONCLUSIONS: We found a higher risk of AMI related to surgical removal of the tonsils and appendix before age 20. These results are consistent with the hypothesis that subtle alterations in immune function following these operations may alter the subsequent cardiovascular risk, but further studies are needed to confirm these findings and to explore possible mechanisms. PMID- 21632601 TI - Mother's body size and placental size predict coronary heart disease in men. AB - AIMS: People whose birthweights were towards the lower end of the normal range are at increased risk of coronary heart disease. This is attributed to foetal programming through malnutrition, but the cause of the malnutrition is unknown. METHODS AND RESULTS: We studied 6975 men born in Helsinki during 1934-44. Their size at birth was recorded. Babies who later developed coronary heart disease tended to have a low ponderal index (birthweight/length(3)). Three different placental phenotypes predicted the disease. In primiparous mothers who were short, having below median height, the hazard ratio for the disease was 1.14 (95% confidence interval 1.08-1.21, P< 0.0001) for each centimetre increase in the difference between the length and breadth of the placental surface. In tall mothers whose body mass index was above the median, the hazard ratio was 1.25 (1.10-1.42, P= 0.0007) per 40 cm(2) decrease in the surface area. In tall mothers whose body mass index was below the median, the hazard ratio was 1.07 (1.02-1.13, P= 0.01) per 1% increase in the placental weight/birthweight ratio. CONCLUSIONS: Three different combinations of maternal and placental size predicted coronary heart disease. The mother's body size determines the availability of nutrients and is linked to the development and function of the placenta, reflected in its shape and size. We speculate that variations in three processes of normal placental development lead to foetal malnutrition. The processes are (i) implantation and spiral artery invasion, (ii) growth of the chorionic surface, and (iii) compensatory expansion of the chorionic surface. PMID- 21632602 TI - Fear of dying and inflammation following acute coronary syndrome. AB - AIMS: Many patients are afraid of dying during acute coronary syndrome (ACS), but the origins and biological correlates of these emotional responses are poorly understood. This study evaluated the prevalence of fear of dying, associations with inflammatory responses during ACS, and later heart rate variability (HRV) and cortisol secretion. METHODS AND RESULTS: Two hundred and eight patients admitted with clinically verified ACS rated their fear of dying on interview in hospital. Plasma tumour necrosis factor (TNF)alpha was recorded on admission, and HRV and salivary cortisol were assessed 3 weeks later. Intense distress and fear of dying was experienced by 21.7%, with moderate levels in 66.1% patients. Fear of dying was more common in younger, lower socioeconomic status, and unmarried patients. It was positively associated with plasma TNFalpha on admission after controlling for sociodemographic factors, clinical risk, and pain intensity (adjusted odds = 4.67, 95% C.I. 1.66-12.65). TNFalpha was associated with reduced HRV 3 weeks later, adjusting for clinical and sociodemographic factors and medication (P = 0.019), while fear of dying was associated with reduced cortisol output (P = 0.004). CONCLUSIONS: Intense distress and fear of dying and heightened inflammation may be related manifestations of an acute biobehavioural response to severe cardiac injury, and have implications for prognostically significant biological risk processes. PMID- 21632603 TI - Heart and mind: are we closer to disentangling the relationship between emotions and poor prognosis in heart disease? PMID- 21632604 TI - Semantic-JSON: a lightweight web service interface for Semantic Web contents integrating multiple life science databases. AB - Global cloud frameworks for bioinformatics research databases become huge and heterogeneous; solutions face various diametric challenges comprising cross integration, retrieval, security and openness. To address this, as of March 2011 organizations including RIKEN published 192 mammalian, plant and protein life sciences databases having 8.2 million data records, integrated as Linked Open or Private Data (LOD/LPD) using SciNetS.org, the Scientists' Networking System. The huge quantity of linked data this database integration framework covers is based on the Semantic Web, where researchers collaborate by managing metadata across public and private databases in a secured data space. This outstripped the data query capacity of existing interface tools like SPARQL. Actual research also requires specialized tools for data analysis using raw original data. To solve these challenges, in December 2009 we developed the lightweight Semantic-JSON interface to access each fragment of linked and raw life sciences data securely under the control of programming languages popularly used by bioinformaticians such as Perl and Ruby. Researchers successfully used the interface across 28 million semantic relationships for biological applications including genome design, sequence processing, inference over phenotype databases, full-text search indexing and human-readable contents like ontology and LOD tree viewers. Semantic JSON services of SciNetS.org are provided at http://semanticjson.org. PMID- 21632605 TI - Effect of a small priming dose on myoclonic movements after intravenous anaesthesia induction with Etomidate-Lipuro in children. AB - BACKGROUND: In children, the incidence of injection pain at i.v. anaesthetic induction with Etomidate-Lipuro is low when compared with propofol mixed with lidocaine (5%). However, the incidence of involuntary myoclonic movements (MM) after induction of anaesthesia is higher compared with propofol (85% vs. 15%). In adults, the incidence of MM is reported to be significantly reduced if a small priming dose is administered immediately before the main injection of etomidate. The aim of this prospective, randomized, double-blind, placebo-controlled clinical trial was to investigate if a small priming dose of etomidate effectively can reduce the incidence of MM also in children. METHODS: Eighty ASA I-II children (1-15 yr) were randomized to receive either a small priming dose of etomidate (0.03 mg kg(-1)) or a lipid emulsion placebo. A standardized induction dose of etomidate (0.3 mg kg(-1)) was administered 60 s after the priming dose. The occurrence and severity (observational score 0-3) of MM was defined as the primary endpoint of the study and was recorded during a 2 min period after induction of anaesthesia. A post hoc analysis was performed regarding the incidence of MM with respect to age. RESULTS: No difference in the occurrence or severity of MM was found between the two study groups, the total incidence of MM being 73.8% (95% confidence interval: 62.7-83.0%). The incidence of MM (score > 0) was found to be statistically higher in the age group 5-10 yr compared with <5 yr; and >10 yr (P=0.0008 and 0.01730, respectively). The MM scores were highest in patients aged 5-10 yr (P=0.0021). CONCLUSIONS: Children in the age range of 5 10 yr appear to be especially prone to react with involuntary MM after i.v. induction of anaesthesia with etomidate. The use of a small, non-sedative, priming dose did not influence the incidence of involuntary MM after i.v. induction of anaesthesia with etomidate in children 1-15 yr of age. PMID- 21632606 TI - Sunscreen SPFs: clear as daylight? PMID- 21632607 TI - Ticagrelor for acute coronary syndrome? AB - Current guidelines from the National Institute for Clinical Excellence (NICE) recommend antiplatelet therapy comprising aspirin plus either clopidogrel or prasugrel for patients with acute coronary syndrome (ACS). However, such dual therapy increases the likelihood of bleeding compared to that with aspirin alone. Ticagrelor (Brilique - Astra-Zeneca) is a new oral antiplatelet drug recently licensed in the UK (since publication of the NICE guidelines) for use with aspirin in patients with ACS, including those managed medically or undergoing percutaneous coronary intervention (PCI) or coronary artery bypass grafting (CABG). Here we review the place of ticagrelor in the management of people with ACS, and whether it offers advantages over standard therapy in terms of greater efficacy or lower likelihood of bleeding complications. PMID- 21632608 TI - Do sunscreens have a role in preventing skin cancer? AB - Solar ultraviolet (UV) radiation is largely responsible for the harmful effects of sun exposure, notably sunburn, photosensitivity and skin cancer. UV radiation is, for example, implicated in malignant melanoma (the most serious form of skin cancer), the incidence of which has risen in the UK over the last 30 years. Such risks make it crucial to know whether measures aimed at reducing sun damage offer worthwhile benefit. Here we assess whether the sunscreen products commonly used to prevent sunburn have a place in the prevention of skin cancer. PMID- 21632609 TI - Assessing Levy walks as models of animal foraging. AB - The hypothesis that the optimal search strategy is a Levy walk (LW) or Levy flight, originally suggested in 1995, has generated an explosion of interest and controversy. Long-standing empirical evidence supporting the LW hypothesis has been overturned, while new models and data are constantly being published. Statistical methods have been criticized and new methods put forward. In parallel with the empirical studies, theoretical search models have been developed. Some theories have been disproved while others remain. Here, we gather together the current state of the art on the role of LWs in optimal foraging theory. We examine the body of theory underpinning the subject. Then we present new results showing that deviations from the idealized one-dimensional search model greatly reduce or remove the advantage of LWs. The search strategy of an LW with exponent MU = 2 is therefore not as robust as is widely thought. We also review the available techniques, and their potential pitfalls, for analysing field data. It is becoming increasingly recognized that there is a wide range of mechanisms that can lead to the apparent observation of power-law patterns. The consequence of this is that the detection of such patterns in field data implies neither that the foragers in question are performing an LW, nor that they have evolved to do so. We conclude that LWs are neither a universal optimal search strategy, nor are they as widespread in nature as was once thought. PMID- 21632610 TI - Probing mechanical principles of focal contacts in cell-matrix adhesion with a coupled stochastic-elastic modelling framework. AB - Cell-matrix adhesion depends on the collective behaviours of clusters of receptor ligand bonds called focal contacts between cell and extracellular matrix. While the behaviour of a single molecular bond is governed by statistical mechanics at the molecular scale, continuum mechanics should be valid at a larger scale. This paper presents an overview of a series of recent theoretical studies aimed at probing the basic mechanical principles of focal contacts in cell-matrix adhesion via stochastic-elastic models in which stochastic descriptions of molecular bonds and elastic descriptions of interfacial traction-separation are unified in a single modelling framework. The intention here is to illustrate these principles using simple analytical and numerical models. The aim of the discussions is to provide possible clues to the following questions: why does the size of focal adhesions (FAs) fall into a narrow range around the micrometre scale? How can cells sense and respond to substrates of varied stiffness via FAs? How do the magnitude and orientation of mechanical forces affect the binding dynamics of FAs? The effects of cluster size, cell-matrix elastic modulus, loading direction and cytoskeletal pretension on the lifetime of FA clusters have been investigated by theoretical arguments as well as Monte Carlo numerical simulations, with results showing that intermediate adhesion size, stiff substrate, cytoskeleton stiffening, low-angle pulling and moderate cytoskeletal pretension are factors that contribute to stable FAs. From a mechanistic point of view, these results provide possible explanations for a wide range of experimental observations and suggest multiple mechanisms by which cells can actively control adhesion and de adhesion via cytoskeletal contractile machinery in response to mechanical properties of their surroundings. PMID- 21632611 TI - Epidemiological game-theory dynamics of chickenpox vaccination in the USA and Israel. AB - The general consensus from epidemiological game-theory studies is that vaccination coverage driven by self-interest (Nash vaccination) is generally lower than group-optimal coverage (utilitarian vaccination). However, diseases that become more severe with age, such as chickenpox, pose an exception to this general consensus. An individual choice to be vaccinated against chickenpox has the potential to harm those not vaccinated by increasing the average age at infection and thus the severity of infection as well as those already vaccinated by increasing the probability of breakthrough infection. To investigate the effects of these externalities on the relationship between Nash and utilitarian vaccination coverages for chickenpox, we developed a game-theory epidemic model that we apply to the USA and Israel, which has different vaccination programmes, vaccination and treatment costs, as well as vaccination coverage levels. We find that the increase in chickenpox severity with age can reverse the typical relationship between utilitarian and Nash vaccination coverages in both the USA and Israel. Our model suggests that to obtain herd immunity of chickenpox vaccination, subsidies or external regulation should be used if vaccination costs are high. By contrast, for low vaccination costs, improving awareness of the vaccine and the potential cost of chickenpox infection is crucial. PMID- 21632612 TI - Malagasy dialects and the peopling of Madagascar. AB - The origin of Malagasy DNA is half African and half Indonesian, nevertheless the Malagasy language, spoken by the entire population, belongs to the Austronesian family. The language most closely related to Malagasy is Maanyan (Greater Barito East group of the Austronesian family), but related languages are also in Sulawesi, Malaysia and Sumatra. For this reason, and because Maanyan is spoken by a population which lives along the Barito river in Kalimantan and which does not possess the necessary skill for long maritime navigation, the ethnic composition of the Indonesian colonizers is still unclear. There is a general consensus that Indonesian sailors reached Madagascar by a maritime trek, but the time, the path and the landing area of the first colonization are all disputed. In this research, we try to answer these problems together with other ones, such as the historical configuration of Malagasy dialects, by types of analysis related to lexicostatistics and glottochronology that draw upon the automated method recently proposed by the authors. The data were collected by the first author at the beginning of 2010 with the invaluable help of Joselina Soafara Nere and consist of Swadesh lists of 200 items for 23 dialects covering all areas of the island. PMID- 21632613 TI - Patterning as a signature of human epidermal stem cell regulation. AB - Understanding how stem cells are regulated in adult tissues is a major challenge in cell biology. In the basal layer of human epidermis, clusters of almost quiescent stem cells are interspersed with proliferating and differentiating cells. Previous studies have shown that the proliferating cells follow a pattern of balanced stochastic cell fate. This behaviour enables them to maintain homeostasis, while stem cells remain confined to their quiescent clusters. Intriguingly, these clusters reappear spontaneously in culture, suggesting that they may play a functional role in stem cell auto-regulation. We propose a model of pattern formation that explains how clustering could regulate stem cell activity in homeostatic tissue through contact inhibition and stem cell aggregation. PMID- 21632614 TI - Agricultural intensification, priming for persistence and the emergence of Nipah virus: a lethal bat-borne zoonosis. AB - Emerging zoonoses threaten global health, yet the processes by which they emerge are complex and poorly understood. Nipah virus (NiV) is an important threat owing to its broad host and geographical range, high case fatality, potential for human to-human transmission and lack of effective prevention or therapies. Here, we investigate the origin of the first identified outbreak of NiV encephalitis in Malaysia and Singapore. We analyse data on livestock production from the index site (a commercial pig farm in Malaysia) prior to and during the outbreak, on Malaysian agricultural production, and from surveys of NiV's wildlife reservoir (flying foxes). Our analyses suggest that repeated introduction of NiV from wildlife changed infection dynamics in pigs. Initial viral introduction produced an explosive epizootic that drove itself to extinction but primed the population for enzootic persistence upon reintroduction of the virus. The resultant within farm persistence permitted regional spread and increased the number of human infections. This study refutes an earlier hypothesis that anomalous El Nino Southern Oscillation-related climatic conditions drove emergence and suggests that priming for persistence drove the emergence of a novel zoonotic pathogen. Thus, we provide empirical evidence for a causative mechanism previously proposed as a precursor to widespread infection with H5N1 avian influenza and other emerging pathogens. PMID- 21632615 TI - The stress of parenthood? Increased glucocorticoids in birds with experimentally enlarged broods. AB - Variation in baseline glucocorticoid (cort) levels can be attributed, at least in part, to differences in energetic demands confronting individuals. Elevated baseline cort levels are routinely interpreted as indicating individuals in poor condition, with low relative fitness. However, when greater reproductive effort increases energetic demands, individuals with high cort might paradoxically be those with the highest fitness. Here, we experimentally test the hypothesis that increased reproductive demand causes increases in baseline cort (the Cort Adaptation hypothesis). We measured maternal baseline cort before and after experimentally enlarging and reducing brood sizes in tree swallows (Tachycineta bicolor). Females with experimentally enlarged broods had greater increases in baseline cort and fledged more offspring than females with reduced broods. Additionally, females with greater increases in baseline cort had higher offspring-provisioning rates than females with lower changes in cort. These findings demonstrate that increased reproductive demand can cause increased baseline cort. As yet, we do not know if these increases in cort cause increased allocation of resources towards reproduction, but the positive relationship between parental behaviour and cort suggests that increased cort does not always interfere with reproductive investment, and might instead facilitate it. PMID- 21632616 TI - Risk factors associated with mortality from white-nose syndrome among hibernating bat colonies. AB - White-nose syndrome (WNS) is a disease responsible for unprecedented mortality in hibernating bats. First observed in a New York cave in 2006, mortality associated with WNS rapidly appeared in hibernacula across the northeastern United States. We used yearly presence-absence data on WNS-related mortality among hibernating bat colonies in the Northeast to determine factors influencing its spread. We evaluated hazard models to test hypotheses about the association between the timing of mortality and colony-level covariates, such as distance from the first WNS-affected site, colony size, species diversity, species composition and type of hibernaculum (cave or mine). Distance to origin and colony size had the greatest effects on WNS hazard over the range of observations; the type of hibernaculum and species composition had weaker effects. The distance effect showed a temporal decrease in magnitude, consistent with the pattern of an expanding epizootic. Large, cave-dwelling bat colonies with high proportions of Myotis lucifugus or other species that seek humid microclimates tended to experience early mortality. Our results suggest that the timing of mortality from WNS is largely dependent on colony location, and large colonies tend to be first in an area to experience high mortality associated with WNS. PMID- 21632617 TI - Ocean acidification erodes crucial auditory behaviour in a marine fish. AB - Ocean acidification is predicted to affect marine ecosystems in many ways, including modification of fish behaviour. Previous studies have identified effects of CO(2)-enriched conditions on the sensory behaviour of fishes, including the loss of natural responses to odours resulting in ecologically deleterious decisions. Many fishes also rely on hearing for orientation, habitat selection, predator avoidance and communication. We used an auditory choice chamber to study the influence of CO(2)-enriched conditions on directional responses of juvenile clownfish (Amphiprion percula) to daytime reef noise. Rearing and test conditions were based on Intergovernmental Panel on Climate Change predictions for the twenty-first century: current-day ambient, 600, 700 and 900 uatm pCO(2). Juveniles from ambient CO(2)-conditions significantly avoided the reef noise, as expected, but this behaviour was absent in juveniles from CO(2)-enriched conditions. This study provides, to our knowledge, the first evidence that ocean acidification affects the auditory response of fishes, with potentially detrimental impacts on early survival. PMID- 21632618 TI - Basal superoxide as a sex-specific immune constraint. AB - There is increasing evidence that reactive oxygen species (ROS), a group of unstable and highly reactive chemical molecules, play a key role in regulating and maintaining life-history trade-offs. Upregulation of ROS in association with immune activation is costly because it may result in an imbalance between pro- and antioxidants and, hence, oxidative damage. Previous research aimed at quantifying this cost has mostly focused on changes in the pro-/antioxidant balance subsequent to an immune response. Here, we test the hypothesis that systemic ROS may constrain immune activation. We show that systemic, pre challenge superoxide (SO) levels are negatively related to the strength of the subsequent immune response towards the mitogen phytohaemagglutinin in male, but not female painted dragon lizards (Ctenophorus pictus). We therefore suggest that systemic SO constrains immune activation in painted dragon males. We speculate that this may be due to sex-specific selection pressures on immune investment. PMID- 21632619 TI - Bacteriophage selection against a plasmid-encoded sex apparatus leads to the loss of antibiotic-resistance plasmids. AB - Antibiotic-resistance genes are often carried by conjugative plasmids, which spread within and between bacterial species. It has long been recognized that some viruses of bacteria (bacteriophage; phage) have evolved to infect and kill plasmid-harbouring cells. This raises a question: can phages cause the loss of plasmid-associated antibiotic resistance by selecting for plasmid-free bacteria, or can bacteria or plasmids evolve resistance to phages in other ways? Here, we show that multiple antibiotic-resistance genes containing plasmids are stably maintained in both Escherichia coli and Salmonella enterica in the absence of phages, while plasmid-dependent phage PRD1 causes a dramatic reduction in the frequency of antibiotic-resistant bacteria. The loss of antibiotic resistance in cells initially harbouring RP4 plasmid was shown to result from evolution of phage resistance where bacterial cells expelled their plasmid (and hence the suitable receptor for phages). Phages also selected for a low frequency of plasmid-containing, phage-resistant bacteria, presumably as a result of modification of the plasmid-encoded receptor. However, these double-resistant mutants had a growth cost compared with phage-resistant but antibiotic susceptible mutants and were unable to conjugate. These results suggest that bacteriophages could play a significant role in restricting the spread of plasmid encoded antibiotic resistance. PMID- 21632620 TI - Age-dependent inbreeding risk and offspring fitness costs in female black grouse. AB - Dispersal is an important mechanism used to avoid inbreeding. However, dispersal may only be effective for part of an individual's lifespan since, post-dispersal individuals that breed over multiple reproductive events may risk mating with kin of the philopatric sex as they age. We tested this hypothesis in black grouse Tetrao tetrix, and show that yearling females never mated with close relatives whereas older females did. However, matings were not with direct kin suggesting that short-distance dispersal to sites containing kin and subsequent overlap of reproductive lifespans between males and females were causing this pattern. Chick mass was lower when kinship was high, suggesting important fitness costs associated with inbred matings. This study shows that increased inbreeding risk might be a widespread yet rarely considered cost of ageing. PMID- 21632621 TI - Memory processing in great apes: the effect of time and sleep. AB - Following encoding, memory remains temporarily vulnerable to disruption. Consolidation refers to offline time-dependent processes that continue after encoding and stabilize, transform or enhance the memory trace. Memory consolidation resulting from sleep has been reported for declarative and non declarative memories in humans. We first investigated the temporal course of memory retrieval in chimpanzees, bonobos and orangutans. We found that the amount of retrieved information was time dependent: apes' performance degraded after 1 and 2 h, stabilized after 4 h, started to increase after 8 and 12 h and fully recovered after 24 h. Second, we show that although memories during wakefulness were highly vulnerable to interference from events similar to those witnessed during the original encoding event, an intervening period of sleep not only stabilized apes' memories into more permanent ones but also protected them against interference. PMID- 21632622 TI - Nutritional geometry: gorillas prioritize non-protein energy while consuming surplus protein. AB - It is widely assumed that terrestrial food webs are built on a nitrogen-limited base and consequently herbivores must compensate through selection of high protein foods and efficient nitrogen retention. Like many folivorous primates, gorillas' diet selection supports this assumption, as they apparently prefer protein-rich foods. Our study of mountain gorillas (Gorilla beringei) in Uganda revealed that, in some periods, carbohydrate-rich fruits displace a large portion of protein-rich leaves in their diet. We show that non-protein energy (NPE) intake was invariant throughout the year, whereas protein intake was substantially higher when leaves were the major portion of the diet. This pattern of macronutrient intake suggests that gorillas prioritize NPE and, to achieve this when leaves are the major dietary item, they over-eat protein. The concentrations of protein consumed in relation to energy when leaves were the major portion of the diet were close to the maximum recommended for humans and similar to high-protein human weight-loss diets. By contrast, the concentrations of protein in relation to energy when gorillas ate fruit-dominated diets were similar to those recommended for humans. Our results question the generality of nitrogen limitation in terrestrial herbivores and provide a fascinating contrast with human macronutrient intake. PMID- 21632623 TI - Shame and honour drive cooperation. AB - Can the threat of being shamed or the prospect of being honoured lead to greater cooperation? We test this hypothesis with anonymous six-player public goods experiments, an experimental paradigm used to investigate problems related to overusing common resources. We instructed the players that the two individuals who were least generous after 10 rounds would be exposed to the group. As the natural antithesis, we also test the effects of honour by revealing the identities of the two players who were most generous. The non-monetary, reputational effects induced by shame and honour each led to approximately 50 per cent higher donations to the public good when compared with the control, demonstrating that both shame and honour can drive cooperation and can help alleviate the tragedy of the commons. PMID- 21632624 TI - Phylogenetic differences of mammalian basal metabolic rate are not explained by mitochondrial basal proton leak. AB - Metabolic rates of mammals presumably increased during the evolution of endothermy, but molecular and cellular mechanisms underlying basal metabolic rate (BMR) are still not understood. It has been established that mitochondrial basal proton leak contributes significantly to BMR. Comparative studies among a diversity of eutherian mammals showed that BMR correlates with body mass and proton leak. Here, we studied BMR and mitochondrial basal proton leak in liver of various marsupial species. Surprisingly, we found that the mitochondrial proton leak was greater in marsupials than in eutherians, although marsupials have lower BMRs. To verify our finding, we kept similar-sized individuals of a marsupial opossum (Monodelphis domestica) and a eutherian rodent (Mesocricetus auratus) species under identical conditions, and directly compared BMR and basal proton leak. We confirmed an approximately 40 per cent lower mass specific BMR in the opossum although its proton leak was significantly higher (approx. 60%). We demonstrate that the increase in BMR during eutherian evolution is not based on a general increase in the mitochondrial proton leak, although there is a similar allometric relationship of proton leak and BMR within mammalian groups. The difference in proton leak between endothermic groups may assist in elucidating distinct metabolic and habitat requirements that have evolved during mammalian divergence. PMID- 21632625 TI - The presence of an embryonic opercular flap in amniotes. AB - The operculum is a large flap consisting of several flat bones found on the side of the head of bony fish. During development, the opercular bones form within the second pharyngeal arch, which expands posteriorly and comes to cover the gill bearing arches. With the evolution of the tetrapods and the assumption of a terrestrial lifestyle, it was believed that the operculum was lost. Here, we demonstrate that an embryonic operculum persists in amniotes and that its early development is homologous with that of teleosts. As in zebrafish, the second pharyngeal arch of the chick embryo grows disproportionately and comes to cover the posterior arches. We show that the developing second pharyngeal arch in both chick and zebrafish embryos express orthologous genes and require shh signalling for caudal expansion. In amniotes, however, the caudal edge of the expanded second arch fuses to the surface of the neck. We have detailed how this process occurs and also demonstrated a requirement for thyroid signalling here. Our results thus demonstrate the persistence of an embryonic opercular flap in amniotes, that its fusion mirrors aspects of amphibian metamorphosis and gives insights into the origin of branchial cleft anomalies in humans. PMID- 21632626 TI - Latitude, elevational climatic zonation and speciation in New World vertebrates. AB - Many biodiversity hotspots are located in montane regions, especially in the tropics. A possible explanation for this pattern is that the narrow thermal tolerances of tropical species and greater climatic stratification of tropical mountains create more opportunities for climate-associated parapatric or allopatric speciation in the tropics relative to the temperate zone. However, it is unclear whether a general relationship exists among latitude, climatic zonation and the ecology of speciation. Recent taxon-specific studies obtained different results regarding the role of climate in speciation in tropical versus temperate areas. Here, we quantify overlap in the climatic distributions of 93 pairs of sister species of mammals, birds, amphibians and reptiles restricted to either the New World tropics or to the Northern temperate zone. We show that elevational ranges of tropical- and temperate-zone species do not differ from one another, yet the temperature range experienced by species in the temperate zone is greater than for those in the tropics. Moreover, tropical sister species tend to exhibit greater similarity in their climatic distributions than temperate sister species. This pattern suggests that evolutionary conservatism in the thermal niches of tropical taxa, coupled with the greater thermal zonation of tropical mountains, may result in increased opportunities for allopatric isolation, speciation and the accumulation of species in tropical montane regions. Our study exemplifies the power of combining phylogenetic and spatial datasets of global climatic variation to explore evolutionary (rather than purely ecological) explanations for the high biodiversity of tropical montane regions. PMID- 21632627 TI - Men with elevated testosterone levels show more affiliative behaviours during interactions with women. AB - Testosterone (T) is thought to play a key role in male-male competition and courtship in many vertebrates, but its precise effects are unclear. We explored whether courtship behaviour in humans is modulated and preceded by changes in T. Pairs of healthy male students first competed in a non-physical contest in which their T levels became elevated. Each participant then had a short, informal interaction with either an unfamiliar man or woman. The sex of the stimulus person did not affect the participants' behaviour overall. However, in interactions with women, those men who had experienced a greater T increase during the contest subsequently showed more interest in the woman, engaged in more self-presentation, smiled more and made more eye contact. No such effects were seen in interactions with other men. This is the first study to provide direct evidence that elevating T during male-male competition is followed by increased affiliative behaviour towards women. PMID- 21632628 TI - Feather corticosterone of a nestling seabird reveals consequences of sex-specific parental investment. AB - Offspring of long-lived species should face costs of parental trade-offs that vary with overall energetic demands encountered by parents during breeding. If sex differences exist in how parents make the trade-off, sex-specific differences may exist in the contribution of each parent to those costs. Adaptations of offspring facing such costs are not well understood, but the hormone corticosterone probably plays a role. We manipulated breeding effort in Cory's shearwaters (Calonectris diomedea) to increase costs to offspring and used an integrated measure of corticosterone from chick feathers to investigate how experimental variation in parental investment influences offspring physiology. Average foraging trip duration and foraging efficiency (FE) of breeding pairs were not related to chick corticosterone, but sex biases in FE were. Adult male investment was more strongly related to chick corticosterone than was female investment. Importantly, we show for the first time suppression of adrenocortical activity in nestling Procellariiform seabirds, and explain how our results indicate an adaptive mechanism invoked by chicks facing increased costs of parental trade-offs. PMID- 21632629 TI - Synchronous attack is advantageous: mixed genotype infections lead to higher infection success in trematode parasites. AB - Co-infecting parasite genotypes typically compete for host resources limiting their fitness. The intensity of such competition depends on whether parasites are reproducing in a host, or using it primarily as a transmission vehicle while not multiplying in host tissues (referred to as 'competition hypothesis'). Alternatively, simultaneous attack and co-infection by several parasite genotypes might facilitate parasite infection because such a diverse attack could present an additional challenge to host immune defence (referred to as 'facilitation hypothesis'). We tested the competition hypothesis by comparing the production of transmission stages (cercariae) from snails infected with one or two genotypes of the trematode Diplostomum pseudospathaceum. We found that cercarial production did not differ between the two groups of snails, suggesting lower per genotype production in double infections, and competition for host resources. Second, we tested the facilitation hypothesis by comparing parasite infection success on fishes (proportion of parasites establishing in the host) using cercariae originating from single-infected snails, double-infected snails and artificial mixtures of the single genotypes. In both cases, we found higher infection success when fishes were challenged with two parasite genotypes instead of one, supporting the facilitation hypothesis. Our results suggest that constraints defining the success of multiple genotype infections in parasites with multiple host life cycles include both between-genotype resource competition in the host and performance of host immune defences against a diverse parasite challenge. PMID- 21632630 TI - Strategic female reproductive investment in response to male attractiveness in birds. AB - Life-history theory predicts that individuals should adjust their reproductive effort according to the expected fitness returns on investment. Because sexually selected male traits should provide honest information about male genetic or phenotypic quality, females may invest more when paired with attractive males. However, there is substantial disagreement in the literature whether such differential allocation is a general pattern. Using a comparative meta-regression approach, we show that female birds generally invest more into reproduction when paired with attractive males, both in terms of egg size and number as well as food provisioning. However, whereas females of species with bi-parental care tend to primarily increase the number of eggs when paired with attractive males, females of species with female-only care produce larger, but not more, eggs. These patterns may reflect adaptive differences in female allocation strategies arising from variation in the signal content of sexually selected male traits between systems of parental care. In contrast to reproductive effort, female allocation of immune-stimulants, anti-oxidants and androgens to the egg yolk was not consistently increased when mated to attractive males, which probably reflects the context-dependent costs and benefits of those yolk compounds to females and offspring. PMID- 21632631 TI - Fishing destabilizes the biomass flow in the marine size spectrum. AB - Fishing impacts on marine food webs are predicted by simulations of a size spectrum community model. In this model, predation is determined by predator and prey size and abundance, and drives predator growth and prey mortality. Fishing amplifies temporal oscillations in the biomass flow. Oscillations appear at lower fishing intensity and have wider amplitude when fishing is selective (removes a narrow size range) and/or when large fish are targeted, than when fishing is more balanced (catching a larger size range) or when small fish are targeted. A novel index of size diversity is developed, and is shown to be sensitive to both fishing intensity and selectivity. To avoid unstable food web dynamics with potential harmful consequences for fisheries, limiting both fishing intensity and selectivity might be an appropriate exploitation strategy. PMID- 21632632 TI - Crushing artifacts resulting in small blue cellular clusters that simulate small cell carcinoma. AB - The presence of "crushing" artifacts in histological sections is a very common finding and represents a potentially major pitfall for the surgical pathologist, particularly in small biopsy specimens. Microscopically, it appears as bluish clusters in which the cellular details are not recognizable. Here the authors report examples of this phenomenon that have been reported or that they have personally observed and emphasize some diagnostic clue to avoid its overinterpretation and the dire clinical consequences that this may entail. PMID- 21632633 TI - Small cell lung carcinoma with marked bronchial epithelial involvement. AB - Marked involvement of bronchial epithelium by malignant cells with a neuroendocrine immunophenotype was observed in a pulmonary lobectomy specimen containing combined small cell lung carcinoma (SCLC). Review of the medical literature reveals scant information on malignant neuroendocrine cells in bronchial epithelium accompanying SCLC and no documentation of an SCLC precursor. We discuss the possibility that the intraepithelial neoplastic lesion that we have described may be a primary lesion and possibly a precursor of SCLC and the alternative possibility that it represents invasion by underlying invasive SCLC. The need for further comprehensive study of the morphology and immunophenotype of bronchial mucosal abnormalities accompanying SCLC utilizing lung resection specimens is emphasized. PMID- 21632634 TI - "The Glass Menagerie": with apologies to Tennessee Williams. PMID- 21632635 TI - Bence-Jones protein-type myeloma with amyloid myopathy presenting as amyloidomas and extensive amyloid deposits in the muscularis propria: a rapidly fatal autopsy case. AB - This study reports a 59-year-old man who suffered from multiple skeletal muscle amyloidomas and showed a rapidly fatal course. He noticed left inguinal pain and gait disturbance due to muscle weakness of the left leg. Protein in urine (3.3 g/d) and Bence-Jones protein of the kappa type (2.3 g/d) were detected. Bone marrow aspiration showed 11.6% monoclonal plasma cells in nucleated cells. A core needle-biopsied and resected left inguinal tumor showed the deposition of eosinophilic amorphous materials positive for Congo red stain and the kappa-light chain. He was diagnosed with plasma cell myeloma with AL (amyloid light chain) amyloidosis. Multiple soft-part tumors developed, grew rapidly, and he died 3 months after admission. At autopsy, 3 large amyloidomas were observed in the skeletal muscles, and prominent amyloid deposits were also seen in the diaphragm, intercostal muscle, iliopsoas muscle, and cervical skeletal muscles examined. Massive amyloid materials deposited diffusely in the propria muscularis of the gastrointestinal tract: the tongue to the rectum. PMID- 21632636 TI - Mullerianosis of the urinary bladder. AB - Mullerianosis of the urinary bladder is a rare condition that encompasses 3 histological entities (endometriosis, endocervicosis, and endosalpingiosis). The authors report 2 patients with bladder mullerianosis, one of whom had endometriosis and the other a history of past pelvic surgery, describe the histological and cystoscopic features, and review the literature on this condition. Mucosal hyperemia and submucosal nodules or cysts with associated fibrosis and distortion of the bladder wall may mimic malignancy on cystoscopy, as may the infiltrative growth pattern sometimes evident histologically. Recognition of this complex diagnostic entity is important to avoid misdiagnosis and inappropriate investigation. There is a clinical association with endometriosis and past pelvic surgery. PMID- 21632637 TI - Triple synchronous primary gynecologic carcinomas: a case report and review of the literature. AB - Double synchronous primaries are known to occasionally occur in gynecologic cancers. Cases of triple or quadruple synchronous primaries, with only 4 case reports in the literature, are extremely rare. The authors report the case of a 49-year-old para 2-0-0-2 woman who presented for surgical management of metastatic ovarian adenocarcinoma diagnosed at an outside institution. On examination of the surgical specimen, 3 synchronous primary carcinomas with multiple histologic features were identified within the ovaries, uterus, and cervix. Although rare, the possibility of triple synchronous primary malignancies should be considered when evaluating gynecologic malignancies. PMID- 21632638 TI - Prostate involvement by intravascular large B-cell lymphoma: a case report with literature review. AB - Prostate involvement by intravascular large B-cell lymphoma (IVLBL) is extremely rare. Until now, only 6 cases have been reported in the literature. In this article, the authors report a case of a 65-year-old Chinese man with IVLBL located in the prostate. The diagnosis of IVLBL was obtained incidentally from a biopsy of his prostatectomy specimen. The patient underwent CHOP therapy for 5 cycles and had a favorable clinical outcome. PMID- 21632639 TI - Mesothelin and GPR30 staining among a spectrum of pancreatic epithelial neoplasms. AB - INTRODUCTION: Our study attempts to characterize mesothelin and GPR30 / estrogen receptor (ER) staining in pancreatic pathology. MATERIALS AND METHODS: Immunohistochemical staining for mesothelin, GPR30, and ER was performed on a variety of pancreatic lesions. RESULTS: 24 of 42 (57%) adenocarcinomas stained for mesothelin, while 0 of 16 non-carcinomas (0%) stained (p = 0.0000784). 35 of 39 (90%) adenocarcinomas stained for GPR30, while only 4 of 15 (27%) non carcinomas stained (p = 0.0000036). Apart from stromal staining in one case of mucinous cystic neoplasm, no cases stained for ER. 27 of 37 (73%) adenocarcinoma fine needle aspirates were positive for mesothelin. DISCUSSION: GPR30 is more sensitive, but less specific than mesothelin for pancreatic adenocarcinoma. Mesothelin is detected in most adenocarcinoma fine needle aspirates. ER is rarely detected in pancreatic lesions. PMID- 21632640 TI - Peritoneal bile granulomas identified at Cesarean section and mimicking disseminated malignancy. AB - A 37-year-old woman was found to have multiple peritoneal nodules, including on the uterine serosa, at Cesarean section. Histology showed yellow crystalline material surrounded by histiocytes and occasional multinucleate giant cells. Further enquiry revealed that the patient underwent a cholecystectomy for a perforated gallbladder 3 years earlier. Pathologists should be aware of this uncommon complication of bile leakage, which has the potential to clinically mimic disseminated malignancy. This may become increasingly common in the future since bile leakage is more likely to occur with laparoscopic than open cholecystectomy. PMID- 21632641 TI - Extranodal Rosai-Dorfman disease arising in the right atrium: a case report with literature review. AB - Rosai-Dorfman disease is a rare, benign histiocytic proliferative disorder that commonly affects the lymph nodes. Although extranodal involvement has been reported in diverse sites, manifestation in the cardiovascular system is extremely uncommon. Specifically, the involvement of the heart by Rosai-Dorfman disease is an extraordinarily infrequent event. Here, the authors present a case of Rosai-Dorfman disease arising in the right atrium in a symptomatic 61-year-old man who initially presented with pleuritic chest pain and was found to have a large, lobulated, and circumscribed right atrial mass. The lesion exhibited an exuberant histiocytic and chronic fibroinflammatory process with focal emperipolesis within histiocytes. Immunohistochemical studies demonstrated strong S100 positivity in CD68+ CD1a- histiocytes. Although rare, Rosai-Dorfman disease should be considered in the differential diagnosis of a right atrial mass. PMID- 21632642 TI - Wagner-Meissner corpuscle proliferation: a unique cause of colon polyps. PMID- 21632643 TI - Diagnosis of gastrointestinal stromal tumor (GIST) on transurethral resection of the prostate: a case report and review of the literature. AB - Gastrointesinal stromal tumors (GISTs) are the most common mesenchymal neoplasms of the gastrointestinal tract and they show differentiation towards interstitial cells of Cajal (ICC). Herein, we describe a case of a 60-year-old man presenting with symptoms mimicking adenomatous hyperplasia of the prostate that was subjected to repeated transurethral prostatectomies. Histologic and immunohistochemical examination showed a neoplastic process with characteristics consistent with GIST. Imaging studies confirmed a rectal origin of the tumor. Review of the literature revealed 20 cases of GISTs occurring in the prostate gland, either diagnosed as either primary GISTs or, more commonly, as rectal neoplasms extending to this organ. We add our case in this short list, emphasizing on the importance of inclusion of GISTs in the differential diagnosis of every spindle cell lesion encountered in the prostate. PMID- 21632644 TI - The calcaneal fat pad and the preservation of human dignity. PMID- 21632645 TI - Invited commentary. PMID- 21632646 TI - Invited commentary. PMID- 21632647 TI - Cortical deficits of glutamic acid decarboxylase 67 expression in schizophrenia: clinical, protein, and cell type-specific features. AB - OBJECTIVE: Cognitive deficits in schizophrenia are associated with altered activity of the dorsolateral prefrontal cortex, which has been attributed to lower expression of the 67 kDa isoform of glutamic acid decarboxylase (GAD67), the major gamma-aminobutyric acid (GABA)-synthesizing enzyme. However, little is known about the relationship of prefrontal GAD67 mRNA levels and illness severity, translation of the transcript into protein, and protein levels in axon terminals, the key site of GABA production and function. METHOD: Quantitative polymerase chain reaction was used to measure GAD67 mRNA levels in postmortem specimens of dorsolateral prefrontal cortex from subjects with schizophrenia and matched comparison subjects with no known history of psychiatric or neurological disorders (N=42 pairs). In a subset of this cohort in which potential confounds of protein measures were controlled (N=19 pairs), Western blotting was used to quantify tissue levels of GAD67 protein in tissue. In five of these pairs, multilabel confocal immunofluorescence was used to quantify GAD67 protein levels in the axon terminals of parvalbumin-containing GABA neurons, which are known to have low levels of GAD67 mRNA in schizophrenia. RESULTS: GAD67 mRNA levels were significantly lower in schizophrenia subjects (by 15%), but transcript levels were not associated with predictors or measures of illness severity or chronicity. In schizophrenia subjects, GAD67 protein levels were significantly lower in total gray matter (by 10%) and in parvalbumin axon terminals (by 49%). CONCLUSIONS: The findings that lower GAD67 mRNA expression is common in schizophrenia, that it is not a consequence of having the illness, and that it leads to less translation of the protein, especially in the axon terminals of parvalbumin-containing neurons, support the hypothesis that lower GABA synthesis in parvalbumin neurons contributes to dorsolateral prefrontal cortex dysfunction and impaired cognition in schizophrenia. PMID- 21632648 TI - Common abnormalities and disorder-specific compensation during implicit regulation of emotional processing in generalized anxiety and major depressive disorders. AB - OBJECTIVE: Anxiety and depressive disorders are both associated with abnormalities in the processing and regulation of emotion. However, little is known about the similarities and differences between anxiety and depression at the neural level. The authors examined emotional conflict processing using a salient stimulus associated with observable and interpretable behavioral outcomes and with activation in limbic and prefrontal regions implicated in anxiety and depression. METHOD: Thirty-two healthy comparison subjects, 18 patients with generalized anxiety disorder only, 14 patients with major depression only, and 25 patients with comorbid generalized anxiety disorder and major depression were studied using functional MRI while they performed an emotional conflict task that involved categorizing facial affect while ignoring overlaid affect label words. The authors used behavioral and neural measures to compare trial-by-trial changes in conflict regulation, a test of implicit regulation of emotional processing. RESULTS: Behavioral data indicated that only patients with generalized anxiety (i.e., the anxiety-only and comorbid groups) failed to implicitly regulate emotional conflict. By contrast, deficits in activation and connectivity of the ventral anterior cingulate and amygdala, areas previously implicated in regulating emotional conflict, were found in all patient groups. Depression-only patients, however, compensated for this deficit by also activating the left and right anterior lateral prefrontal cortices, in which activity was correlated with behavioral evidence of successful implicit regulation, thus mediating the disorder-specificity of the behavioral phenotype. CONCLUSIONS: These data support the existence of a common abnormality in anxiety and depression in the ventral cingulate and the amygdala, which may be related to a shared genetic etiology. Compensatory engagement of cognitive control circuitry in depression illustrates how the complex nature of psychopathology arises from the interaction of deficits and compensation, all of which can occur at an implicit level. PMID- 21632649 TI - Progressive gray matter loss and changes in cognitive functioning associated with exposure to herpes simplex virus 1 in schizophrenia: a longitudinal study. AB - OBJECTIVE: Longitudinal changes in gray matter volume and cognitive performance were evaluated among individuals exposed to neurotropic herpes simplex virus subtype 1 (HSV1). There is a replicable association of HSV1 exposure with smaller prefrontal volumes and cognitive impairments in schizophrenia. METHOD: The authors concurrently examined the whole-brain longitudinal trajectory over 1 year of gray matter volumes and executive functioning measured with the Wisconsin Card Sorting Test among 26 first-episode antipsychotic-naive subjects with schizophrenia and 38 healthy subjects. Age, gender, socioeconomic status, and exposure to cytomegalovirus (another virus of the herpes family that was previously associated with cognitive impairments) were the covariates. RESULTS: Significant gray matter loss in the posterior cingulate gyrus was noted among the HSV1-seropositive schizophrenia subjects over 1 year but not among other groups. Prefrontal gray matter volumes did not show longitudinal changes. Binomial mixed effects models indicated that improvement over 1 year in Wisconsin Card Sorting Test categories completed and perseverative errors occurred in significantly fewer HSV1-seropositive schizophrenia subjects than in the HSV1-seronegative schizophrenia subjects or the healthy subjects regardless of serological status. Three-way interactions of diagnosis, HSV1 status, and time were significant for both categories completed and perseverative errors. An increase in perseverative errors over 1 year, but not the change in the number of categories completed, correlated with longitudinal volume loss of the posterior cingulate gyrus. CONCLUSIONS: These observations suggest that HSV1 exposure may be associated with longitudinal gray matter loss in the posterior cingulate gyrus and decline in executive functioning among subjects with schizophrenia. PMID- 21632650 TI - The pathology of social phobia is independent of developmental changes in face processing. AB - OBJECTIVE: While social phobia in adolescence predicts the illness in adulthood, no study has directly compared the neural responses in social phobia in adults and adolescents. The authors examined neural responses to facial expressions in adults and adolescents with social phobia to determine whether the neural correlates of adult social phobia during face processing also manifest in adolescent social phobia. METHOD: Blood-oxygen-level-dependent (BOLD) responses were compared in 39 medication-free participants with social phobia (25 adults and 14 adolescents) and 39 healthy comparison subjects (23 adults and 16 adolescents) matched on age, IQ, and gender. During fMRI scans, participants saw angry, fearful, and neutral expression stimuli while making a gender judgment. RESULTS: Significant diagnosis-by-emotion interactions were observed within the amygdala and the rostral anterior cingulate cortex, as has previously been hypothesized. In these regions, both the adolescent and adult social phobia patients showed significantly increased BOLD responses relative to their respective age-matched comparison subjects, and there was no evidence of age related modulation of between-group differences. These enhanced responses occurred specifically when viewing angry (rostral anterior cingulate cortex) and fearful (amygdala and rostral anterior cingulate cortex) expressions but not when viewing neutral expressions. In addition, the severity of social phobia was significantly correlated with the enhanced rostral anterior cingulate cortex response in the adults. CONCLUSIONS: The neural correlates of adult social phobia during face processing also manifest in adolescents. Neural correlates that are observed in adult social phobia may represent the persistence of profiles established earlier in life rather than adaptive responses to such earlier perturbations or maturational changes. These cross-sectional observations might encourage longitudinal fMRI studies of adolescent social phobia. PMID- 21632651 TI - A pilot study to evaluate assisted freehand ultrasound elasticity imaging in the sizing of early breast cancer: a comparison of B-mode and AFUSON elasticity ultrasound with histopathology measurements. AB - OBJECTIVE: This pilot study investigates the role of assisted-freehand ultrasound (AFUSON) elasticity imaging of the breast in assessing the contour, size and area of 23 early breast cancers by making comparison of AFUSON with the equivalent B mode ultrasound images and gold standard histopathology slides. METHODS: The B mode, AFUSON and digitised histopathology slides of three early breast cancers were compared for contour, size and area with histopathology scans. AFUSON features that corresponded to areas of known malignant change on the histopathology slides were regarded as diagnostic. These diagnostic criteria were then applied to the B-mode and AFUSON elasticity images of all 23 breast cancers in the pilot study without having the availability of the histopathology scans for reference. Corresponding diameters were measured and the results were compared with the equivalent measurements on the scans of the histology slides. The results were tabulated in histogram form. Diagnostic confidence levels were evaluated. RESULTS: Size dimension accuracy increased from 66% using B-mode alone to 82% using combined B-mode and AFUSON elasticity images. Tumour area accuracy was also increased. A small number of cases had a striking visual similarity of shape on AFUSON elasticity scans and histopathology slides. CONCLUSION: In spite of the shortfalls in this study, AFUSON elasticity imaging was capable of acquiring some high-quality images that showed strong correlation between AFUSON elasticity and scans of histology slides. Further studies will be carried out to refine the technique and determine if it has a role in the diagnosis and management of breast cancer. PMID- 21632652 TI - Effects of virtual reality training on functional reaching movements in people with Parkinson's disease: a randomized controlled pilot trial. AB - OBJECTIVE: To investigate whether practising reaching for virtual moving targets would improve motor performance in people with Parkinson's disease. DESIGN: Randomized pretest-posttest control group design. SETTING: A virtual reality laboratory in a university setting. PARTICIPANTS: Thirty-three adults with Parkinson's disease. INTERVENTIONS: The virtual reality training required 60 trials of reaching for fast-moving virtual balls with the dominant hand. The control group had 60 practice trials turning pegs with their non-dominant hand. MAIN OUTCOME MEASURES: Pretest and posttest required reaching with the dominant hand to grasp real stationary balls and balls moving at different speeds down a ramp. Success rates and kinematic data (movement time, peak velocity and percentage of movement time for acceleration phase) from pretest and posttest were recorded to determine the immediate transfer effects. RESULTS: Compared with the control group, the virtual reality training group became faster (F = 9.08, P = 0.005) and more forceful (F = 9.36, P = 0.005) when reaching for real stationary balls. However, there was no significant difference in success rate or movement kinematics between the two groups when reaching for real moving balls. CONCLUSION: A short virtual reality training programme improved the movement speed of discrete aiming tasks when participants reached for real stationary objects. However, the transfer effect was minimal when reaching for real moving objects. PMID- 21632655 TI - Altered turnover of beta-carotene and Chl a in Arabidopsis leaves treated with lincomycin or norflurazon. AB - Interactions between beta-carotene (beta-C) and Chl a turnover were investigated in relation to photoinhibition and D1 protein turnover in mature leaves of Arabidopsis (Arabidopsis thaliana) by 14CO2 pulse-chase labeling. Following a 2 h treatment of leaves with water, lincomycin (Linco; an inhibitor of chloroplast protein synthesis) or norflurazon (NF; an inhibitor of carotenoid biosynthesis at phytoene desaturation) in the dark, 14CO2 was applied to the leaves for 30 min under control light (CL; 130 MUmol photons m-2 s-1) conditions, followed by exposure to either CL or high light (HL; 1,100 MUmol photons m-2 s-1) in ambient CO2 for up to 6 h. Under both light conditions, 14C incorporation was strongly decreased for Chl a and moderately suppressed for beta-C in Linco-treated leaves, showing a marked decline of PSII efficiency (F(v)/F(m)) and beta-C content compared with water-treated leaves. Partial inhibition of carotenoid biosynthesis by NF caused no or only a minor decrease in F(v)/F(m) and Chl a turnover under both conditions, while the beta-C content significantly declined and high 14C labeling was found for phytoene, the substrate of phytoene desaturase. Together, the results suggest coordinated turnover of Chl a and D1, but somewhat different regulation for beta-C turnover, in Arabidopsis leaves. Inhibition of carotenoid biosynthesis by NF may initially enhance metabolic flux in the pathway upstream of phytoene, presumably compensating for short supply of beta-C. Our observations are also in line with the notion that HL-induced accumulation of xanthophylls may involve a precursor pool which is distinct from that for beta-C turnover. PMID- 21632656 TI - Characterization of the selenite uptake mechanism in the coccolithophore Emiliania huxleyi (Haptophyta). AB - The marine coccolithophore Emiliania huxleyi (Haptophyta) requires selenium as an essential element for growth, and the active species absorbed is selenite, not selenate. This study characterized the selenite uptake mechanism using 75Se as a tracer. Kinetic analysis of selenite uptake showed the involvement of both active and passive transport processes. The active transport was suppressed by 0.5 mM vanadate, a membrane-permeable inhibitor of H+-ATPase, at pH 8.3. When the pH was lowered from 8.3 to 5.3, the selenite uptake activity greatly increased, even in the presence of vanadate, suggesting that the H+ concentration gradient may be a motive force for selenite transport. [75Se]Selenite uptake at selenite-limiting concentrations was hardly affected by selenate, sulfate and sulfite, even at 100 MUM. In contrast, 3 MUM orthophosphate increased the K(m) 5-fold. These data showed that HSeO3-, a dominant selenite species at acidic pH, is the active species for transport through the plasma membrane and transport is driven by DeltapH energized by H+-ATPase. Kinetic analysis showed that the selenite uptake activity was competitively inhibited by orthophosphate. Furthermore, the active selenite transport mechanism was shown to be induced de novo under Se-deficient conditions and induction was suppressed by the addition of either sufficient selenite or cycloheximide, an inhibitor of de novo protein synthesis. These results indicate that E. huxleyi cells developed an active selenite uptake mechanism to overcome the disadvantages of Se limitation in ecosystems, maintaining selenium metabolism and selenoproteins for high viability. PMID- 21632657 TI - Spatial and temporal expression of actin depolymerizing factors ADF7 and ADF10 during male gametophyte development in Arabidopsis thaliana. AB - The actin cytoskeleton plays a crucial role in many aspects of plant cell development. During male gametophyte development, the actin arrays are conspicuously remodeled both during pollen maturation in the anther and after pollen hydration on the receptive stigma and pollen tube elongation. Remodeling of actin arrays results from the highly orchestrated activities of numerous actin binding proteins (ABPs). A key player in actin remodeling is the actin depolymerizing factor (ADF), which increases actin filament treadmilling rates. We prepared fluorescent protein fusions of two Arabidopsis pollen-specific ADFs, ADF7 and ADF10. We monitored the expression and subcellular localization of these proteins during male gametophyte development, pollen germination and pollen tube growth. ADF7 and ADF10 were differentially expressed with the ADF7 signal appearing in the microspore stage and that of ADF10 only during the polarized microspore stage. ADF7 was associated with the microspore nucleus and the vegetative nucleus of the mature grain during less metabolically active stages, but in germinating pollen grains and elongating pollen tubes, it was associated with the subapical actin fringe. On the other hand, ADF10 was associated with filamentous actin in the developing gametophyte, in particular with the arrays surrounding the apertures of the mature pollen grain. In the shank of elongating pollen tubes, ADF10 was associated with thick actin cables. We propose possible specific functions of these two ADFs based on their differences in expression and localization. PMID- 21632658 TI - Poor accountability under reforms could damage NHS, warns King's Fund. PMID- 21632659 TI - A chance to optimise "value" in the NHS. PMID- 21632660 TI - Keep GPs in the driving seat. PMID- 21632661 TI - No news is good news: confidential enquiries into maternal deaths. PMID- 21632662 TI - Why sorry doesn't need to be the hardest word. PMID- 21632663 TI - Bad medicine: digital rectal examination. PMID- 21632664 TI - Overcrowding in emergency departments and adverse outcomes. PMID- 21632665 TI - Association between waiting times and short term mortality and hospital admission after departure from emergency department: population based cohort study from Ontario, Canada. AB - OBJECTIVE: To determine whether patients who are not admitted to hospital after attending an emergency department during shifts with long waiting times are at risk for adverse events. DESIGN: Population based retrospective cohort study using health administrative databases. Setting High volume emergency departments in Ontario, Canada, fiscal years 2003-7. PARTICIPANTS: All emergency department patients who were not admitted (seen and discharged; left without being seen). OUTCOME MEASURES: Risk of adverse events (admission to hospital or death within seven days) adjusted for important characteristics of patients, shift, and hospital. RESULTS: 13,934,542 patients were seen and discharged and 617,011 left without being seen. The risk of adverse events increased with the mean length of stay of similar patients in the same shift in the emergency department. For mean length of stay >= 6 v <1 hour the adjusted odds ratio (95% confidence interval) was 1.79 (1.24 to 2.59) for death and 1.95 (1.79 to 2.13) for admission in high acuity patients and 1.71 (1.25 to 2.35) for death and 1.66 (1.56 to 1.76) for admission in low acuity patients). Leaving without being seen was not associated with an increase in adverse events at the level of the patient or by annual rates of the hospital. CONCLUSIONS: Presenting to an emergency department during shifts with longer waiting times, reflected in longer mean length of stay, is associated with a greater risk in the short term of death and admission to hospital in patients who are well enough to leave the department. Patients who leave without being seen are not at higher risk of short term adverse events. PMID- 21632667 TI - A case of recurrent ventricular tachycardia. PMID- 21632666 TI - Longer term effects of very low energy diet on obstructive sleep apnoea in cohort derived from randomised controlled trial: prospective observational follow-up study. AB - OBJECTIVE: To determine whether initial improvements in obstructive sleep apnoea after a very low energy diet were maintained after one year in patients with moderate to severe obstructive sleep apnoea. DESIGN: Single centre, prospective observational follow-up study. SETTING: Outpatient obesity clinic in a university hospital in Stockholm, Sweden. PARTICIPANTS: 63 men aged 30-65 with body mass index (BMI) 30-40 and moderate to severe obstructive sleep apnoea defined as an apnoea-hypopnoea index >= 15 (events/hour), all treated with continuous positive airway pressure. INTERVENTION: A one year weight loss programme, consisting of an initial very low energy diet for nine weeks (seven weeks of 2.3 MJ/day and two weeks of gradual introduction of normal food) followed by a weight loss maintenance programme. MAIN OUTCOME MEASURE: Apnoea-hypopnoea index, the main index for severity of obstructive sleep apnoea. Data from all patients were analysed (baseline carried forward for missing data). RESULTS: Of 63 eligible patients, 58 completed the very low energy diet period and started the weight maintenance programme and 44 completed the full programme; 49 had complete measurements at one year. At baseline the mean apnoea-hypopnoea index was 36 events/hour. After the very low energy diet period, apnoea-hypopnoea index was improved by -21 events/hour (95% confidence interval -17 to -25) and weight by 18 kg (-16 to -19; both P<0.001). After one year the apnoea-hypopnoea index had improved by -17 events/hour (-13 to -21) and body weight by -12 kg (-10 to -14) compared with baseline (both P<0.001). Patients with severe obstructive sleep apnoea at baseline had greater improvements in apnoea-hypopnoea index (-25 events/hour) compared with patients with moderate disease (-7 events/hour, P<0.001). At one year, 30/63 (48%, 95% confidence interval 35% to 60%) no longer required continuous positive airway pressure and 6/63 (10%, 2% to 17%) had total remission of obstructive sleep apnoea (apnoea-hypopnoea index <5 events/hour). There was a dose-response association between weight loss and apnoea-hypopnoea index at follow-up (beta = 0.50 events/kg, 0.11 to 0.88; P = 0.013). CONCLUSION: Initial improvements in obstructive sleep apnoea after treatment with a very low energy diet can be maintained after one year in obese men with moderate to severe disease. Those who lose the most weight or have severe sleep apnoea at baseline benefit most. Trial registration Current Controlled Trials 70090382. PMID- 21632668 TI - Coronal computed tomography of the chest. PMID- 21632669 TI - Involve specialists in commissioning, urges Royal College of Physicians. PMID- 21632670 TI - Focus interventions on preventing HIV on adolescents, says report. PMID- 21632671 TI - GPs and nurses launch end of life care charter. PMID- 21632672 TI - Radiation fears prompt possible restrictions on wi-fi and mobile phone use in schools. PMID- 21632673 TI - Coronary flow reserve and asymmetric dimethylarginine levels: new measurements for identifying subclinical atherosclerosis in patients with psoriatic arthritis. AB - OBJECTIVE: To identify the presence of subclinical atherosclerosis in patients with psoriatic arthritis (PsA) and healthy controls using intima-media thickness (IMT), coronary flow reserve (CFR), and the plasma concentration of asymmetric dimethylarginine (ADMA), to evaluate the correlations among ADMA, IMT, and CFR. METHODS: The study involved 22 patients who fulfilled the ClASsification of Psoriatic ARthritis study group criteria for PsA and a cohort of 35 healthy controls with no history or current signs of coronary artery disease (CAD). Common carotid IMT was measured using high-resolution B-mode ultrasonography. Dipyridamole transthoracic stress echocardiography was used to evaluate CFR. Blood samples were obtained to assess ADMA levels. The clinical manifestations were recorded. All patients were treated with disease-modifying antirheumatic drug, but none had received any biological or steroid therapy. RESULTS: Plasma ADMA levels were significantly higher in the patients with PsA (0.71 +/- 0.07 MUmol/l vs 0.48 +/- 0.07 MUmol/l; p = 0.00) and CFR was significantly reduced in that group (2.86 +/- 0.70 vs 3.3 +/- 0.43; p < 0.01) compared to controls. Common carotid IMT was greater in the patients with PsA, but the difference was not significant (0.64 +/- 0.26 mm vs 0.62 +/- 0.5 mm; p = 0.65). There was a significant correlation between CFR and plasma ADMA levels in the PsA group (R = 0.28; p < 0.01), but no correlation between plasma ADMA levels and IMT (R = 0.02; p = 0.32), Disease Activity Score 28 (p = 0.52), or Psoriasis Area and Severity Index (p = 0.98). CONCLUSION: Our patients with PsA showed a profile of subclinical atherosclerosis. ADMA may be a useful marker of endothelial dysfunction in PsA. PMID- 21632674 TI - Smoking and outcomes after knee and hip arthroplasty: a systematic review. AB - OBJECTIVE: Studies have suggested higher rates of perioperative and postoperative complications in smokers compared to nonsmokers. The objective of this systematic review was to assess the association of smoking and postoperative outcomes following total hip arthroplasty (THA) or total knee arthroplasty (TKA). METHODS: A search of 6 databases (The Cochrane Library, Scopus, Proquest Dissertation abstracts, CINAHL, Ovid Medline, and Embase) was performed by a Cochrane librarian. All titles and abstracts were screened by 2 independent reviewers with expertise in performing systematic reviews. Studies were included if they were fully published reports that included smoking and any perioperative or postoperative clinical outcome in patients with TKA or THA. RESULTS: A total of 21 studies were included for the review, of which 6 provided multivariable adjusted analyses, 14 univariate analyses, and one statistical modeling. For most outcomes, results from 1-2 studies could be pooled. Current smokers were significantly more likely to have any postoperative complication (risk ratio 1.24, 95% CI 1.01 to 1.54) and death (risk ratio 1.63, 95% CI 1.06 to 2.51) compared to nonsmokers. Former smokers were significantly more likely to have any post-operative complication (risk ratio 1.32, 95% CI 1.05 to 1.66) and death (risk ratio 1.69, 95% CI 1.08 to 2.64) compared to nonsmokers. CONCLUSION: This systematic review found that smoking is associated with significantly higher risk of postoperative complication and mortality following TKA or THA. Studies examining longterm consequences of smoking on implant survival and complications are needed. Smoking cessation may improve outcomes after THA or TKA. PMID- 21632675 TI - Secondary osteoporosis in men and women: clinical challenge of an unresolved issue. AB - OBJECTIVE: To evaluate the clinical and etiological factors of osteoporosis. We also tested the FRAX algorithm to compare the assessment of fracture risk in patients with primary or secondary osteoporosis. METHODS: A prospective study carried out in a large sample of 123 men and 246 women. All subjects had a biochemical, densitometric, and radiological examination of thoracic and lumbar spine. RESULTS: The prevalence of primary (men 52.9% vs women 50%; p = nonsignificant) and secondary (men 21.1% vs women 17.5%; p = nonsignificant) osteoporosis did not differ between the sexes. In contrast, the prevalence of primary osteoporosis was significantly higher than secondary causes (p < 0.0001) in both men and women. While women came to our attention for prevention of osteoporosis, men sought help because of clinical symptoms or disease-related complications, such as fractures. As evaluated by the FRAX tool, patients with osteopenia do not need treatment, in agreement with Italian guidelines. The estimated risk of major osteoporotic and hip fractures was significantly higher in women with secondary osteoporosis compared to men and also compared to women with primary osteoporosis. CONCLUSION: The prevalence of secondary osteoporosis in men is similar to that in women and it is less frequent than commonly reported. In patients with secondary osteoporosis, FRAX calculation may provide an estimate of a particularly high fracture risk in patients whose bone fragility is usually attributed to another disease. PMID- 21632676 TI - Differential features between primary ankylosing spondylitis and spondylitis associated with psoriasis and inflammatory bowel disease. AB - OBJECTIVE: To describe differential characteristics of axial involvement in ankylosing spondylitis (AS) as compared with that seen in psoriatic arthritis (PsA) and inflammatory bowel disease (IBD) in a cohort of Ibero-American patients. METHODS: This study included 2044 consecutive patients with spondyloarthritis (SpA; ESSG criteria). Demographic, clinical, disease activity, functional ability, quality of life, work status, radiologic, and therapeutic data were evaluated and collected by RESPONDIA members from different Ibero American countries between June and December 2006. Patients selected for analysis met modified New York criteria (mNY) for AS. RESULTS: A total of 1264 patients met the New York criteria for AS: 1072 had primary AS, 147 had psoriatic, and 45 had IBD-associated spondylitis. Median disease duration was comparable among the 3 patient groups. Patients with primary AS were significantly younger (p = 0.01) and presented a higher frequency of males (p = 0.01) than the other 2 groups. Axial manifestations such as inflammatory back pain and sacroiliac pain were significantly more frequent in patients with primary AS (p = 0.05) versus other groups, whereas frequency of dactylitis, enthesitis, and peripheral arthritis was more common in patients with psoriatic spondylitis (p = 0.05). Spinal mobility was significantly more limited in patients with primary AS versus the other 2 groups (p = 0.0001). Radiologic changes according to BASRI total score were equally significant in primary AS. Disease activity (BASDAI), functional ability (BASFI), and quality of life (ASQoL) scores were comparable in the 3 groups. CONCLUSION: Patients with primary AS had more severe axial involvement than those with spondylitis associated with psoriasis or IBD. Functional capacity, disease activity, and quality of life were comparable among the groups studied. PMID- 21632677 TI - Serum MMP-3 level as a biomarker for monitoring and predicting response to etanercept treatment in ankylosing spondylitis. AB - OBJECTIVE: To investigate whether level of serum matrix metalloproteinase-3 (MMP 3) can serve as a biomarker for monitoring and predicting response to etanercept treatment in patients with ankylosing spondylitis (AS) in daily clinical practice. METHODS: Ninety-two consecutive AS outpatients with active disease who started etanercept treatment were included in this longitudinal observational study. Clinical data were collected prospectively at baseline and after 3 and 12 months of treatment. At the same timepoints, serum MMP-3 levels were measured retrospectively by ELISA. RESULTS: Since baseline serum MMP-3 levels were significantly higher in male compared to female patients with AS, data analysis was split for gender. Changes in serum MMP-3 levels after etanercept treatment correlated positively with changes in clinical assessments of disease activity and physical function in both male and female patients. Receiver operating characteristic analysis in male patients showed that baseline serum MMP-3 levels had poor accuracy (AUC < 0.7) to discriminate between Assessments in Ankylosing Spondylitis 20 (ASAS20) or ASAS40 responders and nonresponders after 3 or 12 months of treatment. The accuracy of change in serum MMP-3 levels from baseline to 3 months in predicting response after 3 or 12 months of treatment was poor for ASAS40 (AUC < 0.7) or moderate for ASAS20 (AUC = 0.752 and 0.744, respectively), and was not superior to the accuracy of change in the currently used objective biomarkers, erythrocyte sedimentation rate and C-reactive protein. CONCLUSION: Although significant changes in serum MMP-3 levels were found after etanercept treatment, data analysis indicates that serum MMP-3 levels are not very useful for monitoring and predicting response to etanercept treatment in patients with AS in daily clinical practice. PMID- 21632678 TI - Pain mechanisms in osteoarthritis: understanding the role of central pain and current approaches to its treatment. AB - In this literature review, the mechanisms underlying pain associated with osteoarthritis (OA) are discussed, along with evidence for the efficacy of medications thought to act centrally to relieve OA pain. We survey the cascade of events from inflammation to activation of nociceptive and neuropathic pathways, to the development and maintenance of central and peripheral sensitization. Preclinical and clinical evidence for the sensitization hypothesis is discussed, along with recently identified genetic variations that may increase sensitivity to pain in patients with OA. Evidence is presented for the efficacy of centrally acting analgesics for OA pain (opioids, antiepileptics, tricyclic antidepressants, and serotonin/norepinephrine receptor inhibitors). PMID- 21632679 TI - Bosentan in clinical practice for treating digital and other ischemic ulcers in Spanish patients with systemic sclerosis: IBER-DU cohort study. AB - OBJECTIVE: To describe treatment outcomes and safety experience with bosentan in patients with systemic sclerosis (SSc) and digital ulcers (DU), in a clinical setting in Spain. METHODS: This was a multicenter, noninterventional retrospective cohort study. Data were collected retrospectively from patients with DU, with or without pulmonary arterial hypertension (PAH), who were initiating bosentan therapy in 2003 (n = 26) or 2004 (n = 41) and followed until May 2005. Data were obtained from centers prescribing bosentan. Relevant measures included number of DU, occurrence of new DU, overall DU clinical status (improved, stabilized, worsened), and bosentan-associated adverse events. RESULTS: Sixty-seven patients with SSc and DU or other ulcers were included. PAH was also present in 12 patients (18%). At the start of bosentan treatment, the median number of DU per patient was 3.0. The median change in number of DU was 3.6 and -5.0 at 12 and 24 months, respectively. Sixty-eight percent of the patients did not develop any new DU at 12 months. DU clinical status was reported at 12 months for 22 patients: 18 patients (81.8%) improved and 4 (18.2%) stabilized. The median treatment duration was 13.0 months. The main adverse event was increase of aminotransferase, observed in 5 patients (7%), leading to discontinuation of treatment in 3 patients (4.4%). CONCLUSION: Previously reported results of bosentan efficacy in DU management are reproducible in clinical practice. This efficacy is maintained in the longterm followup. Bosentan treatment was well tolerated and adverse events were comparable with those observed in previous reports. PMID- 21632680 TI - Implantable cardioverter defibrillator prevents sudden cardiac death in systemic sclerosis. AB - OBJECTIVE: Cardiac involvement means a poor prognosis in systemic sclerosis (SSc). Conduction defects and arrhythmias are frequent in patients with SSc, and may result in sudden cardiac death. We tested whether electrophysiologic studies and implantation of cardioverter defibrillators are recommended when ventricular arrhythmias are present. METHOD: A cardioverter defibrillator was implanted in 10 patients with SSc who had heart involvement. RESULT: After 36 months, analysis of the device showed several episodes of ventricular tachycardia in 3 patients, which were promptly reverted by electrical shock delivery. CONCLUSION: In patients with SSc who are affected by ventricular arrhythmias, the implantation of a cardioverter defibrillator may prevent sudden cardiac death. PMID- 21632681 TI - Pregnancy outcomes in systemic lupus erythematosus with and without previous nephritis. AB - OBJECTIVE: To compare rates and predictors of pregnancy complications in mothers with systemic lupus erythematosus (SLE) with and without previous nephritis (PN). METHODS: Retrospective analysis of 107 pregnancies in 83 women with SLE diagnosed prepregnancy. RESULTS: Mothers with PN had higher rates of preterm delivery (< 37/40, 30% vs 11%, p = 0.029) than those without PN. Women with PN had earlier onset of preeclampsia [median 34.5 weeks (IQR 32-37) vs 37.5 weeks (IQR 35-38, p = 0.047)] that was more frequently complicated by preterm delivery (p = 0.02). Risk factors for preeclampsia in women with PN include 10-13 weeks' gestation diastolic blood pressure > 80 mmHg and proteinuria, and prepregnancy estimated glomerular filtration rate (eGFR) < 90 ml/min/1.73 m(2). In women with PN, midtrimester uterine-artery-Doppler notching had low negative predictive value (47%). After 39 months followup, eGFR was stable in women with or without PN. CONCLUSION: In SLE, preterm deliveries are more frequent and preeclampsia occurs earlier in women with PN, but longterm eGFR is preserved. PMID- 21632682 TI - The potential role of interferon-regulatory factor 7 among Taiwanese patients with systemic lupus erythematosus. AB - OBJECTIVE: Type I interferons (IFN), especially IFN-alpha, have been proposed to underlie the pathogenesis of systemic lupus erythematosus (SLE). Members of the IFN regulatory factor (IRF) family, which regulate IFN expression, have been implicated as risk factors for SLE. Our aims were to investigate the expression of IRF7 and its correlation with disease activity and to explore the association in Taiwanese patients between 2 genetic single-nucleotide polymorphisms (SNP) of IRF7 and SLE. METHODS: IRF7 messenger RNA (mRNA) levels were measured in peripheral blood mononuclear cells by real-time reverse transcription polymerase chain reaction in 51 adult patients with SLE and 65 age-matched and sex-matched controls. Their serum IFN-alpha levels were determined by ELISA and the clinical manifestations were recorded at the same time. Two IRF7 SNP, rs1061501 and rs1061502, were examined by genotyping across 92 patients with SLE and 92 age and sex-matched healthy control subjects. RESULTS: Compared with controls, the expression of IRF7 mRNA was significantly increased in patients with SLE and was positively correlated with both the serum level of IFN-alpha and lupus disease activity. The distribution of SNP rs1061501 by genotype (CC, CT, and TT) and by allele (C, T) was significantly different between the SLE and the control group (p = 0.028 for genotype and p = 0.009 for allele). There were no significant differences for SNP rs1061502. CONCLUSION: The results suggest that dysregulation of IRF7 might mediate an excessive production of IFN-alpha, which then exerts a crucial effect on the pathogenesis of human SLE. The IRF7 SNP rs1061501 TT genotype and T allele are enriched in Taiwanese patients with SLE and thus would seem to be associated with an increased risk of developing SLE. PMID- 21632683 TI - Clinical correlates of urolithiasis in ankylosing spondylitis. AB - OBJECTIVE: To determine the association between urolithiasis and syndesmophyte formation and the effect of urolithiasis on ankylosing spondylitis (AS) disease activity. METHODS: In a longitudinal cohort of 504 patients with AS, we conducted an analysis of all patients with AS who have a history of urolithiasis. All patients met the modified New York criteria for AS. Demographics, clinical characteristics, extraarticular features, and comorbidities are systematically recorded in the database. We compared disease activity, functional indices, medical therapy and radiographic damage between AS patients with (Uro+) and without urolithiasis (Uro-) using the modified Stoke Ankylosing Spondylitis Spinal Score (mSASSS). RESULTS: Thirty-eight patients with AS (7.5%) had a history of urolithiasis in our cohort. Seventy-six patients with AS who did not have urolithiasis, matched for age, sex, and ethnicity, were selected as controls. Patients who were Uro+ were more likely to have more functional disability, based on the Bath AS Functional Index (BASFI; mean 5.3 vs 3.6 in control group, p = 0.003). Trends were noted in the Uro+ group toward higher Bath AS Disease Activity Index (BASDAI; mean 4.9 vs 4.0, p = 0.09), more peripheral joint involvement (p = 0.075), and higher frequency of biologic therapy (p = 0.09). No significant difference was detected in mSASSS or the Bath AS Metrology Index (BASMI). Significant association with diabetes mellitus (DM; p = 0.016) and Crohn's disease (p = 0.006) was noted in the Uro+ group. CONCLUSION: Although there is no acceleration of syndesmophyte formation or spinal mobility restriction, more functional disability was detected in the urolithiasis group. The higher risk with concomitant DM or Crohn's disease should alert clinicians to these comorbidities in Uro+ patients with AS. PMID- 21632684 TI - Dramatic repair of joint damage in psoriatic arthritis. PMID- 21632685 TI - ANCA-associated vasculitides and classification: a conundrum solved? PMID- 21632686 TI - The scourge of HIV infection in sub-Saharan Africa -- a rheumatological perspective. PMID- 21632687 TI - Unhelpful criteria sets for "diagnosis" and "assessment of severity" of fibromyalgia. PMID- 21632688 TI - Pattern recognition: a true clinician's pearl in rheumatology. PMID- 21632690 TI - Successful switch of patients with rheumatoid arthritis developing anti-tumor necrosis factor (anti-TNF)-induced lupus to another anti-TNF agent. PMID- 21632691 TI - Role of anti-tumor necrosis factor-alpha blockers in inducing lupus erythematosus tumidus in "rhupus syndrome". PMID- 21632692 TI - Cardiopulmonary arrest after severe anaphylactic reaction to second infusion of infliximab in a patient with ankylosing spondylitis. PMID- 21632693 TI - Association of parvovirus B19 infection with systemic lupus erythematosus: role of Th1 predominance. PMID- 21632694 TI - The impact of hospital volume on the development of infectious complications after elective abdominal aortic surgery in the Medicare population. AB - OBJECTIVE: A relationship exists between higher hospital volume and lower mortality, yet the impact of hospital volume on infectious complications after elective abdominal aortic aneurysm (AAA) repair is unknown. METHODS: The Medicare database (2005-2007) was utilized. Top 10% for volume were categorized as high volume (HV) and compared to low-volume (LV) centers for infectious complications and utilization. RESULTS: A total of 42 155 endovascular aneurysm repair (EVAR) and 17 210 open AAA were identified. Mortality in HV was significantly lower than in LV after EVAR and open AAA. After EVAR, HV had lower than LV rates of overall infection (3.10% vs 3.51%; P = .021), PNA (0.94% vs 1.27%, P = .002), and sepsis (0.31% vs 0.45%; P = .03). Length of stay (LOS) and total hospital charges were significantly lower at HV compared to LV after both EVAR and open AAA. CONCLUSION: For Medicare beneficiaries, undergoing elective AAA repair at hospitals performing higher volume significantly reduced postoperative infectious complications and hospital resource utilization. Further analysis identifying systematic reasons for disparities may offer cost savings and improve outcomes. PMID- 21632695 TI - True giant aneurysm of the tibio-peroneal trunk: case report and review of the literature. PMID- 21632696 TI - Cure frailty models for survival data: application to recurrences for breast cancer and to hospital readmissions for colorectal cancer. AB - Owing to the natural evolution of a disease, several events often arise after a first treatment for the same subject. For example, patients with a primary invasive breast cancer and treated with breast conserving surgery may experience breast cancer recurrences, metastases or death. A certain proportion of subjects in the population who are not expected to experience the events of interest are considered to be 'cured' or non-susceptible. To model correlated failure time data incorporating a surviving fraction, we compare several forms of cure rate frailty models. In the first model already proposed non-susceptible patients are those who are not expected to experience the event of interest over a sufficiently long period of time. The other proposed models account for the possibility of cure after each event. We illustrate the cure frailty models with two data sets. First to analyse time-dependent prognostic factors associated with breast cancer recurrences, metastases, new primary malignancy and death. Second to analyse successive rehospitalizations of patients diagnosed with colorectal cancer. Estimates were obtained by maximization of likelihood using SAS proc NLMIXED for a piecewise constant hazards model. As opposed to the simple frailty model, the proposed methods demonstrate great potential in modelling multivariate survival data with long-term survivors ('cured' individuals). PMID- 21632697 TI - Gynaecological endoscopic evaluation of 4% icodextrin solution: a European, multicentre, double-blind, randomized study of the efficacy and safety in the reduction of de novo adhesions after laparoscopic gynaecological surgery. AB - BACKGROUND: Gynaecological laparoscopic surgery outcomes can be compromised by the formation of de novo adhesions. This randomized, double-blind study was designed to assess the efficacy and safety of 4% icodextrin solution (Adept((r))) in the reduction of de novo adhesion incidence compared to lactated Ringer's solution (LRS). METHODS: Patients undergoing laparoscopic surgery for removal of myomas or endometriotic cysts were treated with randomized solution as an intra operative irrigant and 1l post-operative instillate. De novo adhesion incidence (number of sites with adhesions), severity and extent were independently scored at a second-look procedure and the efficacy of the two solutions compared. The effect of surgical covariates on adhesion formation was also investigated. Initial exploratory analysis of individual anatomical sites of clinical importance was progressed. RESULTS Of 498 patients randomized, 330 were evaluable (160 LRS--75% myomectomy/25% endometriotic cysts; 170 Adept--79% myomectomy/21% endometriotic cysts). At study completion, 76.2% LRS and 77.6% Adept had >= 1 de novo adhesion. The mean (SD) number of de novo adhesions was 2.58 (2.11) for Adept and 2.58 (2.38) for LRS. The treatment effect difference was not significant (P = 0.909). Assessment of surgical covariates identified significant influences on the mean number of de novo adhesions regardless of treatment, including surgery duration (P = 0.048), blood loss in myomectomy patients (P = 0.019), length of uterine incision in myomectomy patients (P < 0.001) and number of suture knots (P < 0.001). There were 15 adverse events considered treatment related in the LRS patients (7.2%) and 18 in the Adept group (8.3%). Of 17 reported serious adverse events (9 LRS; 8 Adept) none were considered treatment related. CONCLUSIONS: The study confirmed the safety of Adept in laparoscopic surgery. The proportion of patients with de novo adhesion formation was considerably higher than previous literature suggested. Overall there was no evidence of a clinical effect but various surgical covariates including surgery duration, blood loss, number and size of incisions, suturing and number of knots were found to influence de novo adhesion formation. The study provides direction for future research into adhesion reduction strategies in site specific surgery. PMID- 21632698 TI - Cerebrospinal venous insufficiency and multiple sclerosis: investigating the truth. PMID- 21632700 TI - Role of higher dietary carbohydrate intake in cataract development. PMID- 21632702 TI - Surface and volume changes in the lens during accommodation. PMID- 21632703 TI - Three-dimensional magnetic imaging of the phakic crystalline lens during accommodation. PMID- 21632705 TI - Frequency-doubling technology and parasol cells. PMID- 21632706 TI - Cold thermoreceptors, unexpected players in tear production and ocular dryness sensations. PMID- 21632707 TI - Peroxisome proliferator-activated receptor gamma-regulated cathepsin D is required for lipid antigen presentation by dendritic cells. AB - It is well established that dendritic cells (DCs) take up, process, and present lipid Ags in complex with CD1d molecules to invariant NKT cells. The lipid activated transcription factor, peroxisome proliferator-activated receptor gamma (PPARgamma), has previously been shown to regulate CD1d expression in human monocyte-derived DCs, providing a link between lipid metabolism and lipid Ag presentation. We report that PPARgamma regulates the expression of a lysosomal protease, cathepsin D (CatD), in human monocyte-derived DCs. Inhibition of CatD specifically reduced the expansion of invariant NKT cells and furthermore resulted in decreased maturation of saposins, a group of lipid transfer proteins required for lysosomal lipid Ag processing and loading. These results reveal a novel mechanism of lipid Ag presentation and identify CatD as a key component of this machinery and firmly place PPARgamma as the transcriptional regulator linking lipid metabolism and lipid Ag processing. PMID- 21632708 TI - Membrane-bound IL-22 after de novo production in tuberculosis and anti Mycobacterium tuberculosis effector function of IL-22+ CD4+ T cells. AB - The role of IL-22-producing CD4(+) T cells in intracellular pathogen infections is poorly characterized. IL-22-producing CD4(+) T cells may express some effector molecules on the membrane, and therefore synergize or contribute to antimicrobial effector function. This hypothesis cannot be tested by conventional approaches manipulating a single IL-22 cytokine at genetic and protein levels, and IL-22(+) T cells cannot be purified for evaluation due to secretion nature of cytokines. In this study, we surprisingly found that upon activation, CD4(+) T cells in Mycobacterium tuberculosis-infected macaques or humans could evolve into T effector cells bearing membrane-bound IL-22 after de novo IL-22 production. Membrane-bound IL-22(+) CD4(+) T effector cells appeared to mature in vivo and sustain membrane distribution in highly inflammatory environments during active M. tuberculosis infection. Near-field scanning optical microscopy/quantum dot based nanoscale molecular imaging revealed that membrane-bound IL-22, like CD3, distributed in membrane and engaged as ~100-200 nm nanoclusters or ~300-600 nm nanodomains for potential interaction with IL-22R. Importantly, purified membrane bound IL-22(+) CD4(+) T cells inhibited intracellular M. tuberculosis replication in macrophages. Our findings suggest that IL-22-producing T cells can evolve to retain IL-22 on membrane for prolonged IL-22 t(1/2) and to exert efficient cell cell interaction for anti-M. tuberculosis effector function. PMID- 21632709 TI - Cellular competition independent of BAFF/B lymphocyte stimulator results in low frequency of an autoreactive clonotype in mature polyclonal B cell compartments. AB - The peripheral B cell prosurvival cytokine BAFF/B lymphocyte stimulator (BLyS) has been proposed to participate in the regulation of immunological tolerance. Selective elimination or reconstitution of B cells expressing transgene-encoded, autoreactive BCRs upon systemic BLyS depletion or supplementation, respectively, was observed in two separate studies. Such findings led to a model positing a higher dependency of autoreactive B cells on BLyS. We tested this model by exploiting two targeted IgH transgenic mice (H chain knock-in [HKI]) that produce large numbers of follicular (FO) B cells that are either weakly or strongly autoreactive with nuclear autoantigens. Even though HKI B cells do not exhibit classical features of anergy, we found that mature, naive, autoreactive HKI B cells are outcompeted for representation in the periphery by a polyclonal B cell population. However, this is not due to a higher dependency of HKI B cells on BLyS for survival. Additionally, excess BLyS does not rescue HKI B cells from selective elimination. These findings suggest that some autoreactive FO B cells can fully develop while in competition with non-autoreactive cells for BLyS, but remain at a competitive disadvantage for other trophic factors that regulate peripheral stability. As such, our data indicate the existence of peripheral tolerance mechanisms that regulate the frequency of autoreactive FO B cells independent of the BLyS pathway. PMID- 21632710 TI - TLR2 signaling and Th2 responses drive Tannerella forsythia-induced periodontal bone loss. AB - Periodontal disease (PD) is a chronic inflammation of the tooth-supporting soft tissue and alveolar bone due to infection by a select group of gram-negative microbes, which leads to tooth loss if untreated. Because mice deficient in CD4(+) cells are resistant to infection-induced alveolar bone loss, Th cells have been implicated in bone-destructive processes during PD. However, the extent to which different Th cell subtypes play roles in pathogenesis or host protection remains to be defined and is likely to vary depending on the dominant microorganism involved. By far, Porphyromonas gingivalis is the best-studied periodontal microbe in PD. Although the gram-negative anaerobe Tannerella forsythia is also a vital contributor to periodontal bone loss, almost nothing is known about immune responses to this organism. Previous studies from our laboratory revealed that T. forsythia induces periodontal bone loss in mice and that this bone loss depends on the bacterially expressed BspA protein. In this study, we showed that T. forsythia activates murine APCs primarily through TLR2 dependent signaling via BspA. Furthermore, T. forsythia infection causes a pronounced Th2 bias, evidenced by T cell expression of IL-5, but not IFN-gamma or IL-17, in draining lymph nodes. Consistently, deficiencies in TLR2 or STAT6 result in resistance to T. forsythia-induced alveolar bone loss. Thus, TLR2 signaling and Th2 cells play pathogenic roles in T. forsythia-induced alveolar bone destruction. PMID- 21632711 TI - Salmonella synthesizing 1-dephosphorylated [corrected] lipopolysaccharide exhibits low endotoxic activity while retaining its immunogenicity. AB - The development of safe live, attenuated Salmonella vaccines may be facilitated by detoxification of its LPS. Recent characterization of the lipid A 1 phosphatase, LpxE, from Francisella tularensis allowed us to construct recombinant, plasmid-free strains of Salmonella that produce predominantly 1 dephosphorylated lipid A, similar to the adjuvant approved for human use. Complete lipid A 1-dephosphorylation was also confirmed under low pH, low Mg(2+) culture conditions, which induce lipid A modifications. LpxE expression in Salmonella reduced its virulence in mice by five orders of magnitude. Moreover, mice inoculated with these detoxified strains were protected against wild-type challenge. Candidate Salmonella vaccine strains synthesizing pneumococcal surface protein A (PspA) were also confirmed to possess nearly complete lipid A 1 dephosphorylation. After inoculation by the LpxE/PspA strains, mice produced robust levels of anti-PspA Abs and showed significantly improved survival against challenge with wild-type Streptococcus pneumoniae WU2 compared with vector-only immunized mice, validating Salmonella synthesizing 1-dephosphorylated lipid A as an Ag-delivery system. PMID- 21632712 TI - Signal regulatory protein alpha regulates the homeostasis of T lymphocytes in the spleen. AB - The molecular basis for formation of lymphoid follicle and its homeostasis in the secondary lymphoid organs remains unclear. Signal regulatory protein alpha (SIRPalpha), an Ig superfamily protein that is predominantly expressed in dendritic cells or macrophages, mediates cell-cell signaling by interacting with CD47, another Ig superfamily protein. In this study, we show that the size of the T cell zone as well as the number of CD4(+) T cells were markedly reduced in the spleen of mice bearing a mutant (MT) SIRPalpha that lacks the cytoplasmic region compared with those of wild-type mice. In addition, the expression of CCL19 and CCL21, as well as of IL-7, which are thought to be important for development or homeostasis of the T cell zone, was markedly decreased in the spleen of SIRPalpha MT mice. By the use of bone marrow chimera, we found that hematopoietic SIRPalpha is important for development of the T cell zone as well as the expression of CCL19 and CCL21 in the spleen. The expression of lymphotoxin and its receptor, lymphotoxin beta receptor, as well as the in vivo response to lymphotoxin beta receptor stimulation were also decreased in the spleen of SIRPalpha MT mice. CD47 deficient mice also manifested phenotypes similar to SIRPalpha MT mice. These data suggest that SIRPalpha as well as its ligand CD47 are thus essential for steady-state homeostasis of T cells in the spleen. PMID- 21632713 TI - Sleep after vaccination boosts immunological memory. AB - Sleep regulates immune functions. We asked whether sleep can influence immunological memory formation. Twenty-seven healthy men were vaccinated against hepatitis A three times, at weeks 0, 8, and 16 with conditions of sleep versus wakefulness in the following night. Sleep was recorded polysomnographically, and hormone levels were assessed throughout the night. Vaccination-induced Th cell and Ab responses were repeatedly monitored for 1 y. Compared with the wake condition, sleep after vaccination doubled the frequency of Ag-specific Th cells and increased the fraction of Th1 cytokine-producing cells in this population. Moreover, sleep markedly increased Ag-specific IgG1. The effects were followed up for 1 y and were associated with high sleep slow-wave activity during the postvaccination night as well as with accompanying levels of immunoregulatory hormones (i.e., increased growth hormone and prolactin but decreased cortisol release). Our findings provide novel evidence that sleep promotes human Th1 immune responses, implicating a critical role for slow-wave sleep in this process. The proinflammatory milieu induced during this sleep stage apparently acts as adjuvant that facilitates the transfer of antigenic information from APCs to Ag-specific Th cells. Like the nervous system, the immune system takes advantage of the offline conditions during sleep to foster adaptive immune responses resulting in improved immunological memory. PMID- 21632714 TI - Migrational guidance of neutrophils is mechanotransduced via high-affinity LFA-1 and calcium flux. AB - Acute inflammation triggers the innate immune response of neutrophils that efficiently traffic from the bloodstream to concentrate at high numbers at the site of tissue infection or wounding. A gatekeeper in this process is activation of beta(2) integrins, which form bond clusters with ICAM-1 on the endothelial surface. These bond clusters serve dual functions of providing adhesive strength to anchor neutrophils under the shear forces of blood flow and directional guidance for cell polarization and subsequent transmigration on inflamed endothelium. We hypothesized that shear forces transmitted through high-affinity LFA-1 facilitates the cooperation with the calcium release-activated channel Orai1 in directing localized cytoskeletal activation and directed migration. By using vascular mimetic microfluidic channels, we observed neutrophil arrest on a substrate of either ICAM-1 or allosteric Abs that stabilize a high- or low affinity conformation of LFA-1. Neutrophils captured via low-affinity LFA-1 did not exhibit intracellular calcium flux, F-actin polymerization, cell polarization, or directional migration under shear flow. In contrast, high affinity LFA-1 provided orientation along a uropod-pseudopod axis that required calcium flux through Orai1. We demonstrate how the shear stress of blood flow can transduce distinct outside-in signals at focal sites of high-affinity LFA-1 that provide contact-mediated guidance for neutrophil emigration. PMID- 21632715 TI - Dichotomous effects of IFN-gamma on dendritic cell function determine the extent of IL-12-driven antitumor T cell immunity. AB - Sustained intratumoral delivery of IL-12 and GM-CSF can overcome tumor immune suppression and promote T cell-dependent eradication of established disease in murine tumor models. However, the antitumor effector response is transient and rapidly followed by a T suppressor cell rebound. The mechanisms that control the switch from an effector to a regulatory response in this model have not been defined. Because dendritic cells (DC) can mediate both effector and suppressor T cell priming, DC activity was monitored in the tumors and the tumor-draining lymph nodes (TDLN) of IL-12/GM-CSF-treated mice. The studies demonstrated that therapy promoted the recruitment of immunogenic DC (iDC) to tumors with subsequent migration to the TDLN within 24-48 h of treatment. Longer-term monitoring revealed that iDC converted to an IDO-positive tolerogenic phenotype in the TDLN between days 2 and 7. Specifically, day 7 DC lost the ability to prime CD8(+) T cells but preferentially induced CD4(+)Foxp3(+) T cells. The functional switch was reversible, as inhibition of IDO with 1-methyl tryptophan restored immunogenic function to tolerogenic DC. All posttherapy immunological activity was strictly associated with conventional myeloid DC, and no functional changes were observed in the plasmacytoid DC subset throughout treatment. Importantly, the initial recruitment and activation of iDC as well as the subsequent switch to tolerogenic activity were both driven by IFN-gamma, revealing the dichotomous role of this cytokine in regulating IL-12-mediated antitumor T cell immunity. PMID- 21632716 TI - Enhanced cancer immunotherapy using STAT3-depleted dendritic cells with high Th1 inducing ability and resistance to cancer cell-derived inhibitory factors. AB - STAT3 signaling constitutes an important negative feedback mechanism for the maintenance of immune homeostasis, a suppressive signal for the Th1 immune response in murine macrophages, and a cancer immune evasion signal in various immune cells. The strategy for STAT3 signal inhibition should be considered, because these features could impede effective cancer immunotherapy. We have evaluated the effects of STAT3 inactivation in dendritic cells (DCs) on immune responses in mice and humans. DCs derived from LysMcre/STAT3(flox/flox) mice displayed higher cytokine production in response to TLR stimulation, activated T cells more efficiently, and were more resistant to the suppression of cytokine production by cancer-derived immunosuppressive factors compared with DCs from control littermates. Antitumor activities of STAT3-depleted and control DCs were compared by intratumoral administration of gp70 Ag peptide-pulsed DCs in the therapeutic MC38 tumor model. Intratumoral administration of STAT3-depleted DCs significantly inhibited MC38 tumor growth of both injected and nontreated remote tumors. The inhibition was accompanied by an increase in gp70-specific T cell response as well as in systemic Th1 immune response. STAT3-depleted human DCs with adenoviral STAT3 short hairpin RNA were also capable of producing more cytokines with TLR stimulation and more resistant to cancer-derived factors, and they induced tumor Ag-specific T cells more efficiently than control DCs. The identified role of DC STAT3 signaling in both in vivo therapeutic tumor models in mice and in vitro-specific T cell induction in humans indicates that STAT3 inactivated DCs may be a promising approach for cancer immunotherapy. PMID- 21632717 TI - Antimicrobial activity of PLUNC protects against Pseudomonas aeruginosa infection. AB - Epithelial antimicrobial activity may protect the lung against inhaled pathogens. The bactericidal/permeability-increasing protein family has demonstrated antimicrobial activity in vitro. PLUNC (palate, lung, and nasal epithelium associated) is a 25-kDa secreted protein that shares homology with bactericidal/permeability-increasing proteins and is expressed in nasopharyngeal and respiratory epithelium. The objective of this study was to determine whether PLUNC can limit Pseudomonas aeruginosa infection in mice. Transgenic mice (Scgb1a1-hPLUNC) were generated in which human PLUNC (hPLUNC) was directed to the airway epithelium with the Scgb1a1 promoter. The hPLUNC protein (hPLUNC) was detected in the epithelium throughout the trachea and bronchial airways and in bronchoalveolar lavage fluid. Bronchoalveolar lavage fluid from transgenic mice exhibited higher antibacterial activity than that from wild type littermates in vitro. After in vivo P. aeruginosa challenge, Scgb1a1-hPLUNC transgenic mice displayed enhanced bacterial clearance. This was accompanied by a decrease in neutrophil infiltration and cytokine levels. More importantly, the overexpressed hPLUNC in Scgb1a1-hPLUNC transgenic mouse airway significantly enhanced mouse survival against P. aeruginosa-induced respiratory infection. These data indicate that PLUNC is a novel antibacterial protein that likely plays a critical role in airway epithelium-mediated innate immune response. PMID- 21632718 TI - A naive-like population of human CD1d-restricted T cells expressing intermediate levels of promyelocytic leukemia zinc finger. AB - Rare CD1d-alpha-galactosylceramide-specific T cells that do not express the invariant Valpha24 chain of human NKT cells were recently identified after expansion in vitro with the lipid Ag, but their phenotype and frequency in vivo and lineage relationship with NKT cells could not be elucidated. By using a CD1d tetramer-based method to enrich these cells from fresh peripheral blood, we demonstrated their naive-like CD62L(high)CD45RO(-)CD4(+) phenotype and relatively high frequency of ~10(-5) in several healthy individuals. Notably, these cells expressed the NKT lineage-specific transcription promyelocytic leukemia zinc finger (PLZF), indicating a developmental relationship with NKT cells and ruling out the possibility that they were conventional MHC-restricted T cells cross reacting against CD1d-alpha-galactosylceramide. Although PLZF is known to direct the effector program of NKT cells, we show in this study that the naive-like cells expressed it at a significantly lower amount than NKT cells. Further, we present mouse studies demonstrating a sharp PLZF expression threshold requirement for induction of the effector phenotype. These findings directly demonstrate in vivo the existence of naive-like CD1d-restricted human T cells marked by intermediate levels of PLZF. PMID- 21632719 TI - Increased cell surface Fas expression is necessary and sufficient to sensitize lung fibroblasts to Fas ligation-induced apoptosis: implications for fibroblast accumulation in idiopathic pulmonary fibrosis. AB - Idiopathic pulmonary fibrosis (IPF) is associated with the accumulation of collagen-secreting fibroblasts and myofibroblasts in the lung parenchyma. Many mechanisms contribute to their accumulation, including resistance to apoptosis. In previous work, we showed that exposure to the proinflammatory cytokines TNF alpha and IFN-gamma reverses the resistance of lung fibroblasts to apoptosis. In this study, we investigate the underlying mechanisms. Based on an interrogation of the transcriptomes of unstimulated and TNF-alpha- and IFN-gamma-stimulated primary lung fibroblasts and the lung fibroblast cell line MRC5, we show that among Fas-signaling pathway molecules, Fas expression was increased ~6-fold in an NF-kappaB- and p38(mapk)-dependent fashion. Prevention of the increase in Fas expression using Fas small interfering RNAs blocked the ability of TNF-alpha and IFN-gamma to sensitize fibroblasts to Fas ligation-induced apoptosis, whereas enforced adenovirus-mediated Fas overexpression was sufficient to overcome basal resistance to Fas-induced apoptosis. Examination of lung tissues from IPF patients revealed low to absent staining of Fas in fibroblastic cells of fibroblast foci. Collectively, these findings suggest that increased expression of Fas is necessary and sufficient to overcome the resistance of lung fibroblasts to Fas-induced apoptosis. Our findings also suggest that approaches aimed at increasing Fas expression by lung fibroblasts and myofibroblasts may be therapeutically relevant in IPF. PMID- 21632720 TI - Human metapneumovirus glycoprotein G inhibits TLR4-dependent signaling in monocyte-derived dendritic cells. AB - Human metapneumovirus (hMPV) is a major cause of upper and lower respiratory infections in children and adults. Recent work from our group demonstrated that hMPV G glycoprotein is an important virulence factor, responsible for inhibiting innate immune responses in airway epithelial cells. Myeloid dendritic cells (DCs) are potent APCs and play a major role in initiating and modulating the innate and adaptive immune responses. In this study, we found that TLR4 plays a major role in hMPV-induced activation of monocyte-derived DCs (moDCs), as downregulation of its expression by small interfering RNA significantly blocked hMPV-induced chemokine and type I IFN expression. Similar results were found in bone marrow derived DCs from TLR4-deficient mice. moDCs infected with a virus lacking G protein expression produced higher levels of cytokines and chemokines compared with cells infected with wild-type virus, suggesting that G protein plays an inhibitory role in viral-induced cellular responses. Specifically, G protein affects TLR4-dependent signaling, as infection of moDCs with recombinant hMPV lacking G protein inhibited LPS-induced production of cytokine and chemokines significantly less than did wild-type virus, and treatment of moDCs with purified G protein resulted in a similar inhibition of LPS-dependent signaling. Our results demonstrate that hMPV G protein plays an important role in inhibiting host innate immune responses, likely affecting adaptive responses too. PMID- 21632721 TI - Ly49D-mediated ITAM signaling in immature thymocytes impairs development by bypassing the pre-TCR checkpoint. AB - Activating and inhibitory NK receptors regulate the development and effector functions of NK cells via their ITAM and ITIM motifs, which recruit protein tyrosine kinases and phosphatases, respectively. In the T cell lineage, inhibitory Ly49 receptors are expressed by a subset of activated T cells and by CD1d-restricted NKT cells, but virtually no expression of activating Ly49 receptors is observed. Using mice transgenic for the activating receptor Ly49D and its associated ITAM signaling DAP12 chain, we show in this article that Ly49D mediated ITAM signaling in immature thymocytes impairs development due to a block in maturation from the double negative (DN) to double positive (DP) stages. A large proportion of Ly49D/DAP12 transgenic thymocytes were able to bypass the pre TCR checkpoint at the DN3 stage, leading to the appearance of unusual populations of DN4 and DP cells that lacked expression of intracellular (ic) TCRbeta-chain. High levels of CD5 were expressed on ic TCRbeta(-) DN and DP thymocytes from Ly49D/DAP12 transgenic mice, further suggesting that Ly49D-mediated ITAM signaling mimics physiological ITAM signaling via the pre-TCR. We also observed unusual ic TCRbeta(-) single positive thymocytes with an immature CD24(high) phenotype that were not found in the periphery. Importantly, thymocyte development was completely rescued by expression of an Ly49A transgene in Ly49D/DAP12 transgenic mice, indicating that Ly49A-mediated ITIM signaling can fully counteract ITAM signaling via Ly49D/DAP12. Collectively, our data indicate that inappropriate ITAM signaling by activating NK receptors on immature thymocytes can subvert T cell development by bypassing the pre-TCR checkpoint. PMID- 21632722 TI - A high-risk population revealed. PMID- 21632723 TI - Medicaid's increasing role in treating youths with autism spectrum disorders. PMID- 21632724 TI - Transformation: recovering mental health treasures. PMID- 21632725 TI - Outcomes of a randomized study of a peer-taught Family-to-Family Education Program for mental illness. AB - OBJECTIVE: The Family-to-Family Education Program (FTF) is a 12-week course offered by the National Alliance on Mental Illness (NAMI) for family members of adults with mental illness. This study evaluated the course's effectiveness. METHODS: A total of 318 consenting participants in five Maryland counties were randomly assigned to take FTF immediately or to wait at least three months for the next available class with free use of any other NAMI supports or community or professional supports. Participants were interviewed at study enrollment and three months later (at course termination) regarding problem- and emotion-focused coping, subjective illness burden, and distress. A linear mixed-effects multilevel regression model tested for significant changes over time between intervention conditions. RESULTS: FTF participants had significantly greater improvements in problem-focused coping as measured by empowerment and illness knowledge. Exploratory analyses revealed that FTF participants had significantly enhanced emotion-focused coping as measured by increased acceptance of their family member's illness, as well as reduced distress and improved problem solving. Subjective illness burden did not differ between groups. CONCLUSIONS: This study provides evidence that FTF is effective for enhancing coping and empowerment of families of persons with mental illness, although not for reducing subjective burden. Other benefits for problem solving and reducing distress are suggested but require replication. PMID- 21632726 TI - Racial and ethnic service use disparities among homeless adults with severe mental illnesses receiving ACT. AB - OBJECTIVE: Case management-based interventions aimed at improving quality of care have the potential to narrow racial and ethnic disparities among people with chronic illnesses. The aim of this study was to assess the equity effects of assertive community treatment (ACT), an evidence-based case management intervention, among homeless adults with severe mental illness. METHODS: This study used baseline, three-, and 12-month data for 6,829 black, Latino, and white adults who received ACT services through the ACCESS study (Access to Community Care and Effective Services and Support). Zero-inflated Poisson random regression models were used to estimate the adjusted probability of use of outpatient psychiatric services and, among service users, the intensity of use. Odds ratios and rate ratios (RRs) were computed to assess disparities at baseline and over time. RESULTS: No disparities were found in probability of use at baseline or over time. Compared with white users, baseline intensity of use was lower for black users (RR=.89; 95% confidence interval [CI]=.83-.96) and Latino users (RR=.65; CI=.52-.81]). Intensity did not change over time for whites, but it did for black and Latino users. Intensity increased for blacks between baseline and three months (RR=1.11, CI=1.06-1.17]) and baseline and 12 months (RR=1.17, CI=1.11-1.22]). Intensity of use dropped for Latinos between baseline and three months (RR=.83, CI=.70-.98). CONCLUSIONS: Receipt of ACT was associated with a reduction in service use disparities for blacks but not for Latinos. Findings suggest that ACT's equity effects differ depending on race-ethnicity. PMID- 21632727 TI - Moving into the fourth decade of ACT. PMID- 21632728 TI - A randomized controlled trial of the illness management and recovery program for persons with schizophrenia. AB - OBJECTIVE: The aim of the study was to evaluate the effects of the illness management and recovery (IMR) program on symptoms and psychosocial functioning of individuals with schizophrenia or schizoaffective disorder in an outpatient setting in Sweden. METHODS: A total of 41 persons with schizophrenia or schizoaffective disorder who were receiving treatment at six psychiatric outpatient rehabilitation centers were randomly assigned to either an IMR group for nine months or to treatment as usual (control condition). Assessments were conducted at baseline, posttreatment (nine months), and follow-up (21 months) and included self-reports and ratings by clinicians (both blind and nonblind to treatment assignment) of illness management, psychiatric symptoms, recovery, coping, quality of life, hospitalization, insight, and suicidal ideation. RESULTS: As measured by self-report and ratings of nonblinded clinicians, IMR program participants demonstrated significantly greater improvement in illness management than participants in the control condition. Ratings of psychiatric symptoms by blinded clinicians using the Psychosis Evaluation Tool for Common Use by Caregivers and self-reported ratings of psychosocial functioning on the Ways of Coping Questionnaire also showed better outcomes than for participants in treatment as usual. A statistically significant decrease in suicidal ideation between baseline and follow-up was found for IMR program participants. CONCLUSIONS: The study supports previous findings and suggests that the IMR program is effective in improving the ability of individuals with schizophrenia to better manage their illness. PMID- 21632729 TI - Potential disparities in the management of schizophrenia in the United States. AB - OBJECTIVE: This study investigated whether outpatient visits to psychiatrists and primary care physicians (family physicians, general internists, or general practitioners) by individuals with schizophrenia differed in antipsychotic medication management and subsequent hospitalization by age, gender, race ethnicity, insurance, rurality, and region. METHODS: Data for the study were from office visit forms completed between 1999 and 2007 by physicians in the National Ambulatory Medical Care Survey and the National Hospital Ambulatory Medical Care Survey. A total of 3,359 outpatient visits by individuals with a diagnosis of schizophrenia were identified. The research team used four logistic regression models to test the relationship of sociodemographic variables to antipsychotic medication management during the visit and to hospitalization after the visit. The four models controlled for available clinical covariates with or without physician specialty in the entire cohort and in the cohort of visits in which patients had no active psychotic symptoms. RESULTS: In at least three of the four models, the research team observed that visits by non-Hispanic black patients had significantly (p<.05) greater odds of involving antipsychotic medication management than visits by non-Hispanic whites (range of odds ratios [ORs] 1.66 to 1.88) and of resulting in hospitalization (range of ORs, 3.52 to 6.95). In all four models, visits by patients who lacked insurance were significantly less likely to result in hospitalization than visits by patients who had private insurance (OR=<.001 in all models). CONCLUSIONS: These findings provide the first national evidence of potential treatment disparities for schizophrenia. Further research is needed to definitively identify disparities and to understand their causes and consequences. PMID- 21632730 TI - Racial and ethnic disparities in postpartum depression care among low-income women. AB - OBJECTIVE: The goal of this study was to characterize racial-ethnic differences in mental health care utilization associated with postpartum depression in a multiethnic cohort of Medicaid recipients. METHODS: In a retrospective cohort study, administrative claims data from New Jersey's Medicaid program were obtained for 29,601 women (13,001 whites, 13,416 blacks, and 3,184 Latinas) who delivered babies between July 2004 and October 2007. Racial-ethnic differences were estimated with logistic regression for initiation of antidepressant medication or outpatient mental health visits within six months of delivery, follow-up (a prescription refill or second visit), and continued mental health care (at least three visits or three filled antidepressant prescriptions within 120 days). RESULTS: Nine percent (N=1,120) of white women initiated postpartum mental health care, compared with 4% (N=568) of black women and 5% (N=162) of Latinas. With analyses controlling for clinical factors, the odds of initiating treatment after delivery were significantly (p<.001) lower for blacks (adjusted odds ratio [AOR]=.43) and Latinas (AOR=.59) compared with whites. Among those who initiated treatment, blacks and Latinas were less likely than whites to receive follow-up treatment (blacks, AOR=.66, p<.001; Latinas, AOR=.67, p<.05) or continued care (blacks, AOR=.81, p=.069; Latinas, AOR=.67, p<.05). Among those who initiated antidepressant treatment, black women and Latinas were less likely than whites to refill a prescription. CONCLUSIONS: There were significant racial ethnic differences in depression-related mental health care after delivery. Suboptimal treatment was prevalent among all low-income women in the study. However, racial and ethnic disparities in the initiation and continuation of postpartum depression care were particularly troubling and warrant clinical and policy attention. PMID- 21632731 TI - Components of cultural competence in three mental health programs. AB - OBJECTIVE: The aim of this study was to identify components of cultural competence in mental health programs developed for cultural groups by community and mental health professionals from these groups. METHODS: Three programs were studied: a prevention program primarily serving African-American and Afro Caribbean youth, a Latino adult acute inpatient unit, and a Chinese day treatment program in a community-based agency. Nine study-trained field researchers used a semistructured instrument that captures program genealogy, structure, processes, and cultural infusion. Program cultural elements were identified from field notes and from individual and group interviews of consumers and staff (N=104). A research-group consensus process with feedback from program staff was used to group elements by shared characteristics into the program components of cultural competence. RESULTS: Components included communication competencies (with use of colloquialisms and accepted forms of address); staff in culturally acceptable roles; culturally framed trust building (such as pairing youths with mentors), stigma reduction, friendly milieus (such as serving culturally familiar foods and playing music popular with the culture), and services; and peer, family, and community involvement (including use of peer counselors and mentors, hosting parent weekends, and linking clients with senior center and community services). CONCLUSIONS: Incorporating these components into any program in which underserved cultural populations are seen is recommended for improving cultural competence. PMID- 21632732 TI - Do empathy and psychological mindedness affect police officers' decision to enter crisis intervention team training? AB - OBJECTIVE: Police officers' voluntary choice to participate in specialized, week long crisis intervention team (CIT) training is generally assumed to be a critical factor in success as a CIT officer. However, issues about self-selection have not been empirically examined. The investigators hypothesized that officers entering CIT training, especially those electing to take it, would have a higher likelihood of exposure to and experience with mental health issues and mental health professionals and greater empathy and psychological mindedness. METHODS: A total of 177 officers-including 109 non-CIT-trained officers, 24 officers assigned to CIT training, and 44 officers who volunteered for CIT training-were assessed before and after their week-long classes (non-CIT officers were enrolled in other courses). Basic sociodemographic characteristics were assessed, and four psychometrically sound measures of empathy and psychological mindedness were administered. Associations between officer status and sociodemographic variables, past exposure to and experience with mental illness issues, and empathy and psychological mindedness were examined. RESULTS: The three groups did not differ on three of four exposure and experience variables; however, self-selected CIT officers were more likely to have prior exposure to mental health issues and professionals. No differences were found between the three groups on measures of empathy or psychological mindedness. One measure of empathy increased significantly after the weeklong CIT training. CONCLUSIONS: Findings did not support the hypothesis that officers self-selecting into CIT training would have greater baseline empathy and psychological mindedness. Although there may be other justifiable reasons to argue for the importance of volunteering, these two traits did not appear to be greater among officers self-selecting into CIT. PMID- 21632733 TI - Comparison of the mental health of female adult film performers and other young women in California. AB - OBJECTIVE: This study compared self-reported mental health status and current depression of female adult film performers and other young women. METHODS: A cross-sectional structured online survey adapted from the California Women's Health Survey (CWHS) was self-administered to a convenience sample of 134 current female adult film performers via the Internet. Bivariate and multivariate analyses were used to compare data for these women with data for 1,773 women of similar ages who responded to the 2007 CWHS. Main outcome measures were self reported mental health status, measured with the Behavioral Risk Factor Surveillance Survey core-instrument quality-of-life questions, and current depression, measured with the Patient Health Questionnaire-8. RESULTS: Performers reported a mean of 7.2 days of poor mental health in the past 30 days, compared with 4.8 days for CWHS respondents, and 33% met criteria for current depression, compared with 13% of CWHS respondents (p<.01). As children, the adult film performers were more likely to have been victims of forced sex (37% compared with 13% of CWHS respondents), to have lived in poverty (24% and 12%), and to have been placed in foster care (21% and 4%) (p<.01). In the past 12 months, 50% of the performers reported living in poverty and 34% reported experiencing domestic violence, compared with 36% and 6%, respectively, of CWHS respondents (p<.01). As adults, 27% had experienced forced sex, compared with 9% of CWHS respondents (p<.01). CONCLUSIONS: Female adult film performers have significantly worse mental health and higher rates of depression than other California women of similar ages. PMID- 21632734 TI - Connections to outpatient mental health care of youths with repeat emergency department visits for psychiatric crises. AB - OBJECTIVE: This study examined connections to outpatient mental health care before and between an initial and subsequent pediatric psychiatric emergency department visit. METHODS: Data for an eight-year period were obtained from the Pediatric Psychiatry Emergency Evaluation Form (PPEEF), which is completed by a child psychiatrist. A total of 338 youths were identified who returned to the emergency department of an urban general hospital within six months of an index visit. Logistic regression models were used to examine the association between having a connection to outpatient mental health care and repeated use of the emergency department. RESULTS: Behavior problems were the presenting complaint for more than 50% of youths at both the index and second visits. Sixty-five percent of youths reported a connection to an outpatient mental health provider at both visits; 9% did not identify a provider at either visit. Eight percent of the youths who reported a connection to community care at their first visit reported no such connection at their second visit. At the second visit, the likelihood of reporting a connection with outpatient care was nearly five times higher among youths who reported such a connection at the index visit. CONCLUSIONS: Continued use of the emergency department despite a connection with outpatient mental health care raises questions about the views of families and providers about the need for emergency services. More research is needed to better understand patterns of care seeking in order to fully inform policy and program development. PMID- 21632736 TI - Behavioral disorders and diabetes-related outcomes among Massachusetts Medicare and Medicaid beneficiaries. AB - OBJECTIVE: The study investigated whether Massachusetts beneficiaries of Medicare, Medicaid, or both programs who have behavioral disorders have higher rates of diabetes-related complications and hospitalizations. METHODS: This was a retrospective study using merged Medicare and Medicaid claims data from Massachusetts in 2004 and 2005. The study included beneficiaries who had type 2 diabetes, who stayed in nursing homes for fewer than 90 days, and who were enrolled in Medicare or Medicaid (or both) for at least ten months during the study period. ICD-9-CM and Current Procedural Terminology codes were used to identify diabetes complications (eye complications, nephropathy, neuropathy, ischemic heart disease, cerebrovascular disease, lower-limb amputations, and diabetes-related hospitalizations). The rates of adverse diabetes outcomes were compared across behavioral disorders as identified by ICD-9-CM diagnoses. While adjusting for case mix, multivariate logistic regressions were performed to compare the odds of adverse diabetes outcomes among people with mental or substance use disorders with those without these disorders. RESULTS: A total of 106,174 individuals met inclusion criteria. Results from adjusted analysis showed a mixed picture of the relationships between behavioral disorders and adverse diabetes outcomes. Although substance use disorders were associated with higher odds of lower-limb amputations and diabetes-related hospitalizations, beneficiaries with schizophrenia or paranoid states had lower odds of adverse diabetes outcomes. CONCLUSIONS: Medicaid and Medicare beneficiaries with alcohol or drug use disorders had higher rates of adverse diabetes outcomes than other groups, whereas beneficiaries with mental disorders had lower rates of diabetes related complications. PMID- 21632735 TI - Mental health service use by children with serious emotional and behavioral disturbance: results from the LAMS study. AB - OBJECTIVE: The study described service utilization among children with emotional and behavioral disorders making a first visit to outpatient mental health clinics in four Midwest cities. METHODS: Data were from the Longitudinal Assessment of Manic Symptoms study. A total of 707 youths aged six to 12 years 11 months and their parents completed diagnostic assessments; demographic information and mental health service use was self-reported. Analyses examined the relationship of demographic variables, diagnoses, impairment, and comorbidity with type and level of services utilized. RESULTS: Utilization was multimodal; half the youths had received outpatient and school services during their lifetime. Factors unrelated to need (age, sex, race, and insurance) were associated with service type. Children with a bipolar spectrum disorder had higher use of inpatient services than those with depressive or disruptive disorders and were more likely to currently use two or more services. More than half of youths with bipolar or depressive disorders had lifetime use of both medication and therapy, whereas youths with a disruptive behavior disorder were more likely to have used only therapy. Impairment and comorbidity were not related to service utilization. CONCLUSIONS: Use of services began at a very young age and occurred in multiple service sectors. Type of service used was related to insurance and race, underscoring the need for ongoing disparities research. Contrary to findings from administrative data analyses, use of medication alone was infrequent. The low rate of use of combination therapy suggests that clinicians and families need to be educated about the effectiveness of multimodal treatment. PMID- 21632737 TI - An online recovery plan program: can peer coaching increase participation? AB - OBJECTIVE: A pilot study evaluated whether the addition of online coaching from a peer specialist increased participation in an online program featuring educational and interactive modules to promote self-management of bipolar disorder. METHODS: A total of 118 participants with bipolar disorder recruited from online and in-person support groups and clinical settings were enrolled in MyRecoveryPlan. Half the group was randomly assigned to receive online coaching support from a certified peer specialist. RESULTS: Participants offered online peer coaching were more likely to return to the program after registration (71% versus 44%, p=.005) and to continue using the program after three weeks (38% versus 9%, p<.001). This pattern was consistent for all program elements. CONCLUSIONS: The addition of online coaching from a peer specialist significantly increased engagement and retention in the program, but further research is needed to evaluate the program's effects on clinical and recovery outcomes. PMID- 21632738 TI - A comparison of phone-based and on-site assessment of fidelity for assertive community treatment in Indiana. AB - OBJECTIVE: This study investigated the reliability and validity of a phone administered fidelity assessment instrument based on the Dartmouth Assertive Community Treatment Scale (DACTS). METHODS: An experienced rater paired with a research assistant without fidelity assessment experience or a consultant familiar with the treatment site conducted phone-based assessments of 23 teams providing assertive community treatment in Indiana. Using the DACTS, consultants conducted on-site evaluations of the programs. RESULTS: The pairs of phone raters revealed high levels of consistency [intraclass correlation coefficient (ICC)=.92] and consensus (mean absolute difference of .07). Phone and on-site assessment showed strong agreement (ICC=.87) and consensus (mean absolute difference of .07) and agreed within .1 scale point, or 2% of the scoring range, for 83% of sites and within .15 scale point for 91% of sites. Results were unaffected by the expertise level of the rater. CONCLUSIONS: Phone-based assessment could help agencies monitor faithful implementation of evidence-based practices. PMID- 21632739 TI - Knowledge and attitudes as predictors of intentions to seek help for and disclose a mental illness. AB - OBJECTIVE: Individuals often choose not to seek help for or disclose their mental illness. This study examined whether having more positive attitudes and more knowledge about mental illness could predict intentions to seek help from a general practitioner and to disclose a mental illness to friends and family members. METHODS: A Department of Health survey in England assessed knowledge about mental illness, attitudes toward people with mental illness, and level of contact with someone with a mental illness among 1,751 adults representative of the general population. RESULTS: With controls for social grade and race ethnicity, intentions to seek help were predicted by better knowledge about mental illness, tolerance and support for community care of mental illness, and older age. Willingness to disclose one's mental illness was associated with better knowledge. CONCLUSIONS: Initiatives that increase knowledge and positive attitudes about mental illness among the general population may improve the extent to which individuals seek help for and disclose a mental illness. PMID- 21632740 TI - Transitions between jail and community-based treatment for individuals with co occurring disorders. AB - OBJECTIVE: This study assessed transitions to community mental health services among individuals with co-occurring disorders upon release from jail. METHODS: Data from jail and public mental health systems in Wayne County, Michigan, were merged to identify 677 individuals diagnosed as having a serious mental illness and substance use disorder who had been jailed a total of 1,774 times over 48 months starting in 2003. RESULTS: Only 33% of incarcerations (N=573) were followed by community-based treatment; 44% (N=803) were followed by treatment during a subsequent incarceration, and 23% (N=398) by no treatment. Generalized estimating equations found that individuals with schizophrenia and substance dependence were the most likely to obtain community treatment. CONCLUSION: By integrating discharge planning, community mental health providers and jails may ensure a continuum of care that facilitates treatment engagement, limits repeated incarcerations, and improves well-being. PMID- 21632741 TI - Excessive antipsychotic dosing in a Canadian outpatient population. PMID- 21632742 TI - Cognitive impairment in mental illness: an SSA blind spot? PMID- 21632743 TI - Stigmatization of mental illness in Nigerian home videos. PMID- 21632744 TI - A high-confidence human plasma proteome reference set with estimated concentrations in PeptideAtlas. AB - Human blood plasma can be obtained relatively noninvasively and contains proteins from most, if not all, tissues of the body. Therefore, an extensive, quantitative catalog of plasma proteins is an important starting point for the discovery of disease biomarkers. In 2005, we showed that different proteomics measurements using different sample preparation and analysis techniques identify significantly different sets of proteins, and that a comprehensive plasma proteome can be compiled only by combining data from many different experiments. Applying advanced computational methods developed for the analysis and integration of very large and diverse data sets generated by tandem MS measurements of tryptic peptides, we have now compiled a high-confidence human plasma proteome reference set with well over twice the identified proteins of previous high-confidence sets. It includes a hierarchy of protein identifications at different levels of redundancy following a clearly defined scheme, which we propose as a standard that can be applied to any proteomics data set to facilitate cross-proteome analyses. Further, to aid in development of blood-based diagnostics using techniques such as selected reaction monitoring, we provide a rough estimate of protein concentrations using spectral counting. We identified 20,433 distinct peptides, from which we inferred a highly nonredundant set of 1929 protein sequences at a false discovery rate of 1%. We have made this resource available via PeptideAtlas, a large, multiorganism, publicly accessible compendium of peptides identified in tandem MS experiments conducted by laboratories around the world. PMID- 21632745 TI - Ethical and practical challenges of sharing data from genome-wide association studies: the eMERGE Consortium experience. AB - In 2007, the National Human Genome Research Institute (NHGRI) established the Electronic MEdical Records and GEnomics (eMERGE) Consortium (www.gwas.net) to develop, disseminate, and apply approaches to research that combine DNA biorepositories with electronic medical record (EMR) systems for large-scale, high-throughput genetic research. One of the major ethical and administrative challenges for the eMERGE Consortium has been complying with existing data sharing policies. This paper discusses the challenges of sharing genomic data linked to health information in the electronic medical record (EMR) and explores the issues as they relate to sharing both within a large consortium and in compliance with the National Institutes of Health (NIH) data-sharing policy. We use the eMERGE Consortium experience to explore data-sharing challenges from the perspective of multiple stakeholders (i.e., research participants, investigators, and research institutions), provide recommendations for researchers and institutions, and call for clearer guidance from the NIH regarding ethical implementation of its data-sharing policy. PMID- 21632746 TI - Epigenetic signatures distinguish multiple classes of enhancers with distinct cellular functions. AB - Epigenetic regulation of gene enhancer elements is important for establishing and maintaining the identity of cells. Gene enhancer elements are thought to exist in either active or poised states distinguishable by chromatin features, but a complete understanding of the regulation of enhancers is lacking. Here, by using mouse embryonic stem cells and their differentiated derivatives, as well as terminally differentiated cells, we report the coexistence of multiple, defined classes of enhancers that serve distinct cellular functions. Specifically, we found that active enhancers can be subclassified based on varying levels of H3K4me1, H3K27ac, and H3K36me3 and the pSer2/5 forms of RNA polymerase II. The abundance of these histone modifications positively correlates with the expression of associated genes and cellular functions consistent with the identity of the cell type. Poised enhancers can also be subclassified based on presence or absence of H3K27me3 and H3K9me3, conservation, genomic location, expression levels of associated genes, and predicted function of associated genes. These findings not only refine the repertoire of histone modifications at both active and poised gene enhancer elements but also raise the possibility that enhancers associated with distinct cellular functions are partitioned based on specific combinations of histone modifications. PMID- 21632747 TI - Coassembly of REST and its cofactors at sites of gene repression in embryonic stem cells. AB - The differentiation of pluripotent embryonic stem cells is regulated by networks of activating and repressing transcription factors that orchestrate determinate patterns of gene expression. With the recent mapping of target sites for many transcription factors, it has been a conundrum that so few of the genes directly targeted by these factors are transcriptionally responsive to the binding of that factor. To address this, we generated genome-wide maps of the transcriptional repressor REST and five of its corepressors in mouse embryonic stem cells. Combining these binding-site maps with comprehensive gene-expression profiling, we show that REST is functionally heterogeneous. Approximately half of its binding sites apparently are nonfunctional, having weaker binding of REST and low recruitment of corepressors. In contrast, the other sites strongly recruit REST and corepressor complexes with varying numbers of components. Strikingly, the latter sites account for almost all observed gene regulation. These data support a model where productive gene repression by REST requires assembly of a multimeric "repressosome" complex, whereas weak recruitment of REST and its cofactors is insufficient to repress gene expression. PMID- 21632748 TI - Reading TE leaves: new approaches to the identification of transposable element insertions. AB - Transposable elements (TEs) are a tremendous source of genome instability and genetic variation. Of particular interest to investigators of human biology and human evolution are retrotransposon insertions that are recent and/or polymorphic in the human population. As a consequence, the ability to assay large numbers of polymorphic TEs in a given genome is valuable. Five recent manuscripts each propose methods to scan whole human genomes to identify, map, and, in some cases, genotype polymorphic retrotransposon insertions in multiple human genomes simultaneously. These technologies promise to revolutionize our ability to analyze human genomes for TE-based variation important to studies of human variability and human disease. Furthermore, the approaches hold promise for researchers interested in nonhuman genomic variability. Herein, we explore the methods reported in the manuscripts and discuss their applications to aspects of human biology and the biology of other organisms. PMID- 21632749 TI - Human dendritic cells infected with the nonpathogenic Mopeia virus induce stronger T-cell responses than those infected with Lassa virus. AB - The events leading to death in severe cases of Lassa fever (LF) are unknown. Fatality seems to be linked to high viremia and immunosuppression, and cellular immunity, rather than neutralizing antibodies, appears to be essential for survival. We previously compared Lassa virus (LV) with its genetically close but nonpathogenic homolog Mopeia virus (MV), which was used to model nonfatal LF. We showed that strong and early activation of antigen-presenting cells (APC) may play a crucial role in controlling infection. Here we developed an in vitro model of dendritic-cell (DC)-T-cell coculture in order to characterize human T-cell responses induced by MV- or LV-infected DCs. Our results show very different responses to infection with LV and MV. MV strongly and durably stimulated CD8(+) and CD4(+) T cells, showing early and high activation, a strong proliferative response, and acquisition of effector and memory phenotypes. Furthermore, robust and functional CD4(+) and CD8(+) cytotoxic T lymphocytes (CTL) were generated. LV, however, induced only weak memory responses. Thus, this study allows an improved understanding of the pathogenesis and immune mechanisms involved in the control of human LV. PMID- 21632750 TI - Differentiated neuroblastoma cells provide a highly efficient model for studies of productive varicella-zoster virus infection of neuronal cells. AB - Varicella-zoster virus (VZV) is a highly species-specific herpesvirus that targets sensory ganglionic neurons. This species specificity has limited the study of many aspects of VZV pathogenesis, including neuronal infection. We report development of a highly efficient neuroblastoma cell model to study productive VZV infection of neuronal cells. We show that differentiation of SH SY5Y neuroblastoma cells yields a homogenous population of neuron-like cells that are permissive to the full VZV replicative cycle. These cells supported productive infection by both laboratory and clinical VZV isolates, including the live varicella vaccine. This model may enable rapid identification of genetic determinants facilitating VZV neurotropism. PMID- 21632751 TI - Poly(C)-binding protein 2 interacts with sequences required for viral replication in the hepatitis C virus (HCV) 5' untranslated region and directs HCV RNA replication through circularizing the viral genome. AB - Sequences in the 5' untranslated region (5'UTR) of hepatitis C virus (HCV) RNA is important for modulating both translation and RNA replication. The translation of the HCV genome depends on an internal ribosome entry site (IRES) located within the 341-nucleotide 5'UTR, while RNA replication requires a smaller region. A question arises whether the replication and translation functions require different regions of the 5'UTR and different sets of RNA-binding proteins. Here, we showed that the 5'-most 157 nucleotides of HCV RNA is the minimum 5'UTR for RNA replication, and it partially overlaps with the IRES. Stem-loops 1 and 2 of the 5'UTR are essential for RNA replication, whereas stem-loop 1 is not required for translation. We also found that poly(C)-binding protein 2 (PCBP2) bound to the replication region of the 5'UTR and associated with detergent-resistant membrane fractions, which are the sites of the HCV replication complex. The knockdown of PCBP2 by short hairpin RNA decreased the amounts of HCV RNA and nonstructural proteins. Antibody-mediated blocking of PCBP2 reduced HCV RNA replication in vitro, indicating that PCBP2 is directly involved in HCV RNA replication. Furthermore, PCBP2 knockdown reduced IRES-dependent translation preferentially from a dual reporter plasmid, suggesting that PCBP2 also regulated IRES activity. These findings indicate that PCBP2 participates in both HCV RNA replication and translation. Moreover, PCBP2 interacts with HCV 5'- and 3'UTR RNA fragments to form an RNA-protein complex and induces the circularization of HCV RNA, as revealed by electron microscopy. This study thus demonstrates the mechanism of the participation of PCBP2 in HCV translation and replication and provides physical evidence for HCV RNA circularization through 5'- and 3'UTR interaction. PMID- 21632753 TI - Adenovirus structural protein IIIa is involved in the serotype specificity of viral DNA packaging. AB - The packaging of the adenovirus (Ad) genome into a capsid displays serotype specificity. This specificity has been attributed to viral packaging proteins, the IVa2 protein and the L1-52/55K protein. We previously found that the Ad17 L1 52/55K protein was not able to complement the growth of an Ad5 L1-52/55K mutant virus, whereas two other Ad17 packaging proteins, IVa2 and L4-22K, could complement the growth of Ad5 viruses with mutations in the respective genes. In this report, we investigated why the Ad17 L1-52/55K protein was not able to complement the Ad5 L1-52/55K mutant virus. We demonstrate that the Ad17 L1-52/55K protein binds to the Ad5 IVa2 protein in vitro and the Ad5 packaging domain in vivo, activities previously associated with packaging function. The Ad17 L1 52/55K protein also associates with empty Ad5 capsids. Interestingly, we find that the Ad17 L1-52/55K protein is able to complement the growth of an Ad5 L1 52/55K mutant virus in conjunction with the Ad17 structural protein IIIa. The same result was found with the L1-52/55K and IIIa proteins of several other Ad serotypes, including Ad3 and Ad4. The Ad17 IIIa protein associates with empty Ad5 capsids. Consistent with the complementation results, we find that the IIIa protein interacts with the L1-52/55K protein in vitro and associates with the viral packaging domain in vivo. These results underscore the complex nature of virus assembly and genome encapsidation and provide a new model for how the viral genome may tether to the empty capsid during the encapsidation process. PMID- 21632752 TI - Neutralizing anti-gH antibody of Varicella-zoster virus modulates distribution of gH and induces gene regulation, mimicking latency. AB - The anti-glycoprotein H (gH) monoclonal antibody (anti-gH-MAb) that neutralizes varicella-zoster virus (VZV) inhibited cell-to-cell infection, resulting in a single infected cell without apoptosis or necrosis, and the number of infectious cells in cultures treated with anti-gH-MAb declined to undetectable levels in 7 to 10 days. Anti-gH-MAb modulated the wide cytoplasmic distribution of gH colocalized with glycoprotein E (gE) to the cytoplasmic compartment with endoplasmic reticulum (ER) and Golgi markers near the nucleus, while gE retained its cytoplasmic distribution. Thus, the disintegrated distribution of gH and gE caused the loss of cellular infectivity. After 4 weeks of treatment with anti-gH MAb, no infectious virus was recovered, even after cultivation without anti-gH MAb for another 8 weeks or various other treatments. Cells were infected with Oka varicella vaccine expressing hepatitis B surface antigen (ROka) and treated with anti-gH-MAb for 4 weeks, and ROka was recovered from the quiescently infected cells by superinfection with the parent Oka vaccine. Among the genes 21, 29, 62, 63, and 66, transcripts of gene 63 were the most frequently detected, and products from the genes 63 and 62, but not gE, were detected mainly in the cytoplasm of quiescently infected cells, in contrast to their nuclear localization in lytically infected cells. The patterns of transcripts and products from the quiescently infected cells were similar to those of latent VZV in human ganglia. Thus, anti-gH-MAb treatment resulted in the antigenic modulation and dormancy of infectivity of VZV. Antigenic modulation by anti-gH MAb illuminates a new aspect in pathogenesis in VZV infection and the gene regulation of VZV during latency in human ganglia. PMID- 21632754 TI - Detection of minority resistance during early HIV-1 infection: natural variation and spurious detection rather than transmission and evolution of multiple viral variants. AB - Reports of a high frequency of the transmission of minority viral populations with drug-resistant mutations (DRM) are inconsistent with evidence that HIV-1 infections usually arise from mono- or oligoclonal transmission. We performed ultradeep sequencing (UDS) of partial HIV-1 gag, pol, and env genes from 32 recently infected individuals. We then evaluated overall and per-site diversity levels, selective pressure, sequence reproducibility, and presence of DRM and accessory mutations (AM). To differentiate biologically meaningful mutations from those caused by methodological errors, we obtained multinomial confidence intervals (CI) for the proportion of DRM at each site and fitted a binomial mixture model to determine background error rates for each sample. We then examined the association between detected minority DRM and the virologic failure of first-line antiretroviral therapy (ART). Similar to other studies, we observed increased detection of DRM at low frequencies (average, 0.56%; 95% CI, 0.43 to 0.69; expected UDS error, 0.21 +/- 0.08% mutations/site). For 8 duplicate runs, there was variability in the proportions of minority DRM. There was no indication of increased diversity or selection at DRM sites compared to other sites and no association between minority DRM and AM. There was no correlation between detected minority DRM and clinical failure of first-line ART. It is unlikely that minority viral variants harboring DRM are transmitted and maintained in the recipient host. The majority of low-frequency DRM detected using UDS are likely errors inherent to UDS methodology or a consequence of error-prone HIV-1 replication. PMID- 21632755 TI - Host response to polyomavirus infection is modulated by RNA adenosine deaminase ADAR1 but not by ADAR2. AB - Adenosine deaminases acting on RNA (ADARs) catalyze the C-6 deamination of adenosine (A) to produce inosine (I), which behaves as guanine (G), thereby altering base pairing in RNAs with double-stranded character. Two genes, adar1 and adar2, are known to encode enzymatically active ADARs in mammalian cells. Furthermore, two size forms of ADAR1 are expressed by alternative promoter usage, a short (p110) nuclear form that is constitutively made and a long (p150) form that is interferon inducible and present in both the cytoplasm and nucleus. ADAR2 is also a constitutively expressed nuclear protein. Extensive A-to-G substitution has been described in mouse polyomavirus (PyV) RNA isolated late times after infection, suggesting modification by ADAR. To test the role of ADAR in PyV infection, we used genetically null mouse embryo fibroblast cells deficient in either ADAR1 or ADAR2. The single-cycle yields and growth kinetics of PyV were comparable between adar1(-/-) and adar2(-/-) genetic null fibroblast cells. While large T antigen was expressed to higher levels in adar1(-/-) cells than adar2(-/ ) cells, less difference was seen in VP1 protein expression levels between the two knockout MEFs. However, virus-induced cell killing was greatly enhanced in PyV-infected adar1(-/-) cells compared to that of adar2(-/-) cells. Complementation with p110 protected cells from PyV-induced cytotoxicity. UV irradiated PyV did not display any enhanced cytopathic effect in adar1(-/-) cells. Reovirus and vesicular stomatitis virus single-cycle yields were comparable between adar1(-/-) and adar2(-/-) cells, and neither reovirus nor VSV showed enhanced cytotoxicity in adar1(-/-)-infected cells. These results suggest that ADAR1 plays a virus-selective role in the host response to infection. PMID- 21632756 TI - Mx is dispensable for interferon-mediated resistance of chicken cells against influenza A virus. AB - The type I interferon (IFN) system plays an important role in antiviral defense against influenza A viruses (FLUAV), which are natural chicken pathogens. Studies of mice identified the Mx1 protein as a key effector molecule of the IFN-induced antiviral state against FLUAV. Chicken Mx genes are highly polymorphic, and recent studies suggested that an Asn/Ser polymorphism at amino acid position 631 determines the antiviral activity of the chicken Mx protein. By employing chicken embryo fibroblasts with defined Mx-631 polymorphisms and retroviral vectors for the expression of Mx isoforms in chicken cells and embryonated eggs, we show here that neither the 631Asn nor the 631Ser variant of chicken Mx was able to confer antiviral protection against several lowly and highly pathogenic FLUAV strains. Using a short interfering RNA (siRNA)-mediated knockdown approach, we noted that the antiviral effect of type I IFN in chicken cells was not dependent on Mx, suggesting that some other IFN-induced factors must contribute to the inhibition of FLUAV in chicken cells. Finally, we found that both isoforms of chicken Mx protein appear to lack GTPase activity, which might explain the observed lack of antiviral activity. PMID- 21632757 TI - Genomic and proteomic analysis of invertebrate iridovirus type 9. AB - Iridoviruses (IV) are nuclear cytoplasmic large DNA viruses that are receiving increasing attention as sublethal pathogens of a range of insects. Invertebrate iridovirus type 9 (IIV-9; Wiseana iridovirus) is a member of the major phylogenetic group of iridoviruses for which there is very limited genomic and proteomic information. The genome is 205,791 bp, has a G+C content of 31%, and contains 191 predicted genes, with approximately 20% of its repeat sequences being located predominantly within coding regions. The repeated sequences include 11 proteins with helix-turn-helix motifs and genes encoding related tandem repeat amino acid sequences. Of the 191 proteins encoded by IIV-9, 108 are most closely related to orthologs in IIV-3 (Chloriridovirus genus), and 114 of the 126 IIV-3 genes have orthologs in IIV-9. In contrast, only 97 of 211 IIV-6 genes have orthologs in IIV-9. There is almost no conservation of gene order between IIV-3, IIV-6, and IIV-9. Phylogenetic analysis using a concatenated sequence of 26 core IV genes confirms that IIV-3 is more closely related to IIV-9 than to IIV-6, despite being from a different genus of the Iridoviridae. An interaction between IIV and small RNA regulatory systems is supported by the prediction of seven putative microRNA (miRNA) sequences combined with XRN exonuclease, RNase III, and double-stranded RNA binding activities encoded on the genome. Proteomic analysis of IIV-9 identified 64 proteins in the virus particle and, when combined with infected cell analysis, confirmed the expression of 94 viral proteins. This study provides the first full-genome and consequent proteomic analysis of group II IIV. PMID- 21632758 TI - The herpes simplex virus 1 UL17 protein is the second constituent of the capsid vertex-specific component required for DNA packaging and retention. AB - The herpes simplex virus (HSV) UL17 and UL25 minor capsid proteins are essential for DNA packaging. They are thought to comprise a molecule arrayed in five copies around each of the capsid vertices. This molecule was initially termed the "C capsid-specific component" (CCSC) (B. L. Trus et al., Mol. Cell 26:479-489, 2007), but as we have subsequently observed this feature on reconstructions of A, B, and C capsids, we now refer to it more generally as the "capsid vertex specific component" (CVSC) (S. K. Cockrell et al., J. Virol. 85:4875-4887, 2011). We previously confirmed that UL25 occupies the vertex-distal region of the CVSC density by visualizing a large UL25-specific tag in reconstructions calculated from cryo-electron microscopy (cryo-EM) images. We have pursued the same strategy to determine the capsid location of the UL17 protein. Recombinant viruses were generated that contained either a small tandem affinity purification (TAP) tag or the green fluorescent protein (GFP) attached to the C terminus of UL17. Purification of the TAP-tagged UL17 or a similarly TAP-tagged UL25 protein clearly demonstrated that the two proteins interact. A cryo-EM reconstruction of capsids containing the UL17-GFP protein reveals that UL17 is the second component of the CVSC and suggests that UL17 interfaces with the other CVSC component, UL25, through its C terminus. The portion of UL17 nearest the vertex appears to be poorly constrained, which may provide flexibility in interacting with tegument proteins or the DNA-packaging machinery at the portal vertex. The exposed locations of the UL17 and UL25 proteins on the HSV-1 capsid exterior suggest that they may be attractive targets for highly specific antivirals. PMID- 21632759 TI - Structure of the three N-terminal immunoglobulin domains of the highly immunogenic outer capsid protein from a T4-like bacteriophage. AB - The head of bacteriophage T4 is decorated with 155 copies of the highly antigenic outer capsid protein (Hoc). One Hoc molecule binds near the center of each hexameric capsomer. Hoc is dispensable for capsid assembly and has been used to display pathogenic antigens on the surface of T4. Here we report the crystal structure of a protein containing the first three of four domains of Hoc from bacteriophage RB49, a close relative of T4. The structure shows an approximately linear arrangement of the protein domains. Each of these domains has an immunoglobulin-like fold, frequently found in cell attachment molecules. In addition, we report biochemical data suggesting that Hoc can bind to Escherichia coli, supporting the hypothesis that Hoc could attach the phage capsids to bacterial surfaces and perhaps also to other organisms. The capacity for such reversible adhesion probably provides survival advantages to the bacteriophage. PMID- 21632760 TI - The 2.3-angstrom structure of porcine circovirus 2. AB - Porcine circovirus 2 (PCV2) is a T=1 nonenveloped icosahedral virus that has had severe impact on the swine industry. Here we report the crystal structure of an N terminally truncated PCV2 virus-like particle at 2.3-A resolution, and the cryo electron microscopy (cryo-EM) image reconstruction of a full-length PCV2 virus like particle at 9.6-A resolution. This is the first atomic structure of a circovirus. The crystal structure revealed that the capsid protein fold is a canonical viral jelly roll. The loops connecting the strands of the jelly roll define the limited features of the surface. Sulfate ions interacting with the surface and electrostatic potential calculations strongly suggest a heparan sulfate binding site that allows PCV2 to gain entry into the cell. The crystal structure also allowed previously determined epitopes of the capsid to be visualized. The cryo-EM image reconstruction showed that the location of the N terminus, absent in the crystal structure, is inside the capsid. As the N terminus was previously shown to be antigenic, it may externalize through viral "breathing." PMID- 21632761 TI - Modulation of TRIM5alpha activity in human cells by alternatively spliced TRIM5 isoforms. AB - TRIM5alpha is a restriction factor that can block an early step in the retroviral life cycle by recognizing and causing the disassembly of incoming viral capsids, thereby preventing the completion of reverse transcription. Numerous other isoforms of human TRIM5 exist, and isoforms lacking a C-terminal SPRY domain can inhibit the activity of TRIM5alpha. Thus, TRIM5alpha activity in a given cell type could be dependent on the relative proportions of TRIM5 isoforms expressed, but little information concerning the relative expression of TRIM5 isoforms in human cells is available. In this study, we demonstrate that mRNAs coding for TRIM5alpha represent only 50% of total TRIM5 transcripts in human cell lines, CD4(+) T cells, and macrophages. Transcripts coding for, in order of abundance, TRIM5iota (TRIM5-iota), a previously uncharacterized isoform, TRIM5gamma, TRIM5delta, and TRIM5kappa are also present. Like TRIM5gamma and TRIM5delta, TRIM5iota and TRIM5kappa do not inhibit HIV-1 replication, but both have dominant negative activity against TRIM5alpha. Specific knockdown of TRIM5iota increases TRIM5alpha activity in human U373-X4 cells, indicating that physiological levels of expression of truncated TRIM5 isoforms in human cells can reduce the activity of TRIM5alpha. PMID- 21632763 TI - Genetic editing of herpes simplex virus 1 and Epstein-Barr herpesvirus genomes by human APOBEC3 cytidine deaminases in culture and in vivo. AB - Human APOBEC3 cytidine deaminases target and edit single-stranded DNA, which can be of viral, mitochondrial, or nuclear origin. Retrovirus genomes, such as human immunodeficiency virus (HIV) genomes deficient in the vif gene and the hepatitis B virus genome, are particularly vulnerable. The genomes of some DNA viruses, such as human papillomaviruses, can be edited in vivo and in transfection experiments. Accordingly, herpesviruses should be no exception. This is indeed the case for herpes simplex virus 1 (HSV-1) in tissue culture, where APOBEC3C (A3C) overexpression can reduce virus titers and the particle/PFU ratio ~10-fold. Nonetheless, A3A, A3G, and AICDA can edit what is presumably a small fraction of HSV genomes in an experimental setting without seriously impacting the viral titer. Hyperediting was found in HSV genomes recovered from 4/8 uncultured buccal lesions. The phenomenon is not restricted to HSV, since hyperedited Epstein-Barr virus (EBV) genomes were readily recovered from 4/5 established cell lines, indicating that episomes are vulnerable to editing. These findings suggest that the widely expressed A3C cytidine deaminase can function as a restriction factor for some human herpesviruses. That the A3C gene is not induced by type I interferons begs the question whether some herpesviruses encode A3C antagonists. PMID- 21632762 TI - Newcastle disease virus-vectored rabies vaccine is safe, highly immunogenic, and provides long-lasting protection in dogs and cats. AB - Effective, safe, and affordable rabies vaccines are still being sought. Newcastle disease virus (NDV), an avian paramyxovirus, has shown promise as a vaccine vector for mammals. Here, we generated a recombinant avirulent NDV La Sota strain expressing the rabies virus glycoprotein (RVG) and evaluated its potential to serve as a vaccine against rabies. The recombinant virus, rL-RVG, retained its high-growth property in chicken eggs, with titers of up to 109.8 50% egg infective doses (EID50)/ml of allantoic fluid. RVG expression enabled rL-RVG to spread from cell to cell in a rabies virus-like manner, and RVG was incorporated on the surface of the rL-RVG viral particle. RVG incorporation did not alter the trypsin-dependent infectivity of the NDV vector in mammalian cells. rL-RVG and La Sota NDV showed similar levels of sensitivity to a neutralization antibody against NDV and similar levels of resistance to a neutralization antibody against rabies virus. Animal studies demonstrated that rL-RVG is safe in several species, including cats and dogs, when administered as multiple high doses of recombinant vaccine. Intramuscular vaccination with rL-RVG induced a substantial rabies virus neutralization antibody response and provided complete protection from challenge with circulating rabies virus strains. Most importantly, rL-RVG induced strong and long-lasting protective neutralization antibody responses to rabies virus in dogs and cats. A low vaccine dose of 108.3 EID50 completely protected dogs from challenge with a circulating strain of rabies virus for more than a year. This is the first study to demonstrate that immunization with an NDV-vectored vaccine can induce long-lasting, systemic protective immunity against rabies. PMID- 21632764 TI - Establishment of the black-tailed prairie dog (Cynomys ludovicianus) as a novel animal model for comparing smallpox vaccines administered preexposure in both high- and low-dose monkeypox virus challenges. AB - The 2003 monkeypox virus (MPXV) outbreak and subsequent laboratory studies demonstrated that the black-tailed prairie dog is susceptible to MPXV infection and that the ensuing rash illness is similar to human systemic orthopoxvirus (OPXV) infection, including a 7- to 9-day incubation period and, likely, in some cases a respiratory route of infection; these features distinguish this model from others. The need for safe and efficacious vaccines for OPVX in areas where it is endemic or epidemic is important to protect an increasingly OPXV-naive population. In this study, we tested current and investigational smallpox vaccines for safety, induction of anti-OPXV antibodies, and protection against mortality and morbidity in two MPXV challenges. None of the smallpox vaccines caused illness in this model, and all vaccinated animals showed anti-OPXV antibody responses and neutralizing antibody. We tested vaccine efficacy by challenging the animals with 10(5) or 10(6) PFU Congo Basin MPXV 30 days postvaccination and evaluating morbidity and mortality. Our results demonstrated that vaccination with either Dryvax or Acambis2000 protected the animals from death with no rash illness. Vaccination with IMVAMUNE also protected the animals from death, albeit with (modified) rash illness. Based on the results of this study, we believe prairie dogs offer a novel and potentially useful small animal model for the safety and efficacy testing of smallpox vaccines in pre- and postexposure vaccine testing, which is important for public health planning. PMID- 21632766 TI - Identification of the first human gyrovirus, a virus related to chicken anemia virus. AB - We have identified in a skin swab sample from a healthy donor a new virus that we have named human gyrovirus (HGyV) because of its similarity to the chicken anemia virus (CAV), the only previously known member of the Gyrovirus genus. In particular, this virus encodes a homolog of the CAV apoptin, a protein that selectively induces apoptosis in cancer cells. By PCR screening, HGyV was found in 5 of 115 other nonlesional skin specimens but in 0 of 92 bronchoalveolar lavages or nasopharyngeal aspirates and in 0 of 92 fecal samples. PMID- 21632765 TI - Impact of intra- and interspecies variation of occludin on its function as coreceptor for authentic hepatitis C virus particles. AB - Hepatitis C virus (HCV) is characterized by a narrow host range and high interindividual variability in the clinical course of infection. Both of these traits are thought to be largely due to genetic variation between species and between individual hosts. The tight junction component occludin (OCLN) is essential for HCV entry into host cells, and the differences between human and murine OCLN are thought to account in part for the inability of HCV to infect mice and hence preclude their use as a convenient small-animal model. This study assesses the impact of genetic variation in OCLN on cell culture-grown HCV (HCVcc) using a newly generated and characterized OCLN(low) subclone of the Huh 7.5 cell line (Huh-7.5 subclone in which endogenous OCLN expression has been downregulated by a short hairpin RNA). We report the frequency of coding nonsynonymous single nucleotide polymorphisms, i.e., polymorphisms resulting in amino acid exchanges, present in the human population and determine their ability to function as HCV (co)receptors. Moreover, we show that murine OCLN can sustain HCVcc entry, albeit with about 5-fold reduced efficiency compared to that of human OCLN. This reduction in efficiency is due solely to two amino acid residues previously identified by others using an HCV pseudoparticle approach. Finally, we use the Huh-7.5/OCLN(low) cell line to show that HCV spread between neighboring cells is strictly dependent on OCLN. PMID- 21632767 TI - A single amino acid in nonstructural protein NS4B confers virulence to dengue virus in AG129 mice through enhancement of viral RNA synthesis. AB - Dengue (DEN) is a mosquito-borne viral disease that has become an increasing economic and health burden for the tropical and subtropical world. The lack of an appropriate animal model of DEN has greatly impeded the study of its pathogenesis and the development of vaccines/antivirals. We recently reported a DEN virus 2 (DENV-2) strain (D2Y98P) that lethally infects immunocompromised AG129 mice, resulting in organ damage or dysfunction and increased vascular permeability, hallmarks of severe DEN in patients (G. K. Tan et al., PLoS Negl. Trop. Dis. 4:e672, 2010). Here we report the identification of one critical virulence determinant of strain D2Y98P. By mutagenesis, we showed that a Phe-to-Leu alteration at amino acid position 52 in nonstructural protein NS4B completely abolished the pathogenicity of the D2Y98P virus, as evidenced by a lack of lethality and the absence of histological signs of disease, which correlated with reduced viral titers and intact vascular permeability. Conversely, a Leu-to-Phe alteration at position 52 of NS4B in nonvirulent DENV-2 strain TSV01 led to 80% lethality and increased viremia. The NS4B(Phe52) viruses displayed enhanced RNA synthesis in mammalian cells but not in mosquito cells. The increased viral RNA synthesis was independent of the ability of NS4B to interfere with the host type I interferon response. Overall, our results demonstrate that Phe at position 52 in NS4B confers virulence in mice on two independent DENV-2 strains through enhancement of viral RNA synthesis. In addition to providing further insights into the functional role of NS4B protein, our findings further support a direct relationship between viral loads and DEN pathogenesis in vivo, consistent with observations in DEN patients. PMID- 21632768 TI - Crimean-Congo hemorrhagic fever virus activates endothelial cells. AB - Crimean-Congo hemorrhagic fever virus (CCHFV) causes viral hemorrhagic fever with high case-fatality rates and is geographically widely distributed. Due to the requirement for a biosafety level 4 (BSL-4) laboratory and the lack of an animal model, knowledge of the viral pathogenesis is limited. Crimean-Congo hemorrhagic fever (CCHF) is characterized by hemorrhage and vascular permeability, indicating the involvement of endothelial cells (ECs). The interplay between ECs and CCHFV is therefore important for understanding the pathogenesis of CCHF. In a previous study, we found that CCHFV-infected monocyte-derived dendritic cells (moDCs) activated ECs; however, the direct effect of CCHFV on ECs was not investigated. Here, we report that ECs are activated upon infection, as demonstrated by upregulation of mRNA levels for E-selectin, vascular cell adhesion molecule 1 (VCAM1), and intercellular adhesion molecule 1 (ICAM1). Protein levels and cell surface expression of ICAM1 responded in a dose-dependent manner to increasing CCHFV titers with concomitant increase in leukocyte adhesion. Furthermore, we examined vascular endothelial (VE) cadherin in CCHFV-infected ECs by different approaches. Infected ECs released higher levels of interleukin 6 (IL-6) and IL-8; however, stimulation of resting ECs with supernatants derived from infected ECs did not result in increased ICAM1 expression. Interestingly, the moDC-mediated activation of ECs was abrogated by addition of neutralizing tumor necrosis factor alpha (TNF-alpha) antibody to moDC supernatants, thereby identifying this soluble mediator as the key cytokine causing EC activation. We conclude that CCHFV can exert both direct and indirect effects on ECs. PMID- 21632769 TI - Protection against rectal transmission of an emtricitabine-resistant simian/human immunodeficiency virus SHIV162p3M184V mutant by intermittent prophylaxis with Truvada. AB - Daily preexposure prophylaxis (PrEP) with Truvada (emtricitabine [FTC] and tenofovir disoproxil fumarate [TDF]) is a novel HIV prevention strategy recently found to reduce HIV incidence among men who have sex with men. We used a macaque model of HIV transmission to investigate if Truvada maintains prophylactic efficacy against an FTC-resistant isolate containing the M184V mutation. Five macaques received a dose of Truvada 3 days before exposing them rectally to the simian/human immunodeficiency virus mutant SHIV162p3(M184V), followed by a second dose 2 h after exposure. Five untreated animals were used as controls. Virus exposures were done weekly for up to 14 weeks. Despite the high (>100-fold) level of FTC resistance conferred by M184V, all five treated animals were protected from infection, while the five untreated macaques were infected (P = 0.0008). Our results show that Truvada maintains high prophylactic efficacy against an FTC resistant isolate. Increased susceptibility to tenofovir due to M184V and other factors, including residual antiviral activity by FTC and/or reduced virus fitness due to M184V, may all have contributed to the observed protection. PMID- 21632770 TI - Shared ancestry between a newfound mole-borne hantavirus and hantaviruses harbored by cricetid rodents. AB - Discovery of genetically distinct hantaviruses in multiple species of shrews (order Soricomorpha, family Soricidae) and moles (family Talpidae) contests the conventional view that rodents (order Rodentia, families Muridae and Cricetidae) are the principal reservoir hosts and suggests that the evolutionary history of hantaviruses is far more complex than previously hypothesized. We now report on Rockport virus (RKPV), a hantavirus identified in archival tissues of the eastern mole (Scalopus aquaticus) collected in Rockport, TX, in 1986. Pairwise comparison of the full-length S, M, and L genomic segments indicated moderately low sequence similarity between RKPV and other soricomorph-borne hantaviruses. Phylogenetic analyses, using maximum-likelihood and Bayesian methods, showed that RKPV shared a most recent common ancestor with cricetid-rodent-borne hantaviruses. Distributed widely across the eastern United States, the fossorial eastern mole is sympatric and syntopic with cricetid rodents known to harbor hantaviruses, raising the possibility of host-switching events in the distant past. Our findings warrant more-detailed investigations on the dynamics of spillover and cross-species transmission of present-day hantaviruses within communities of rodents and moles. PMID- 21632772 TI - Event-related potentials reveal an early advantage for luminance contours in the processing of objects. AB - Detection and identification of objects are the most crucial goals of visual perception. We studied the role of luminance and chromatic information for object processing by comparing performance of familiar, meaningful object contours with those of novel, non-object contours. Comparisons were made between full-color and reduced-color object (or non-object) contours. Full-color stimuli contained both chromatic and luminance information, whereas luminance information was absent in the reduced-color stimuli. All stimuli were made equally salient by fixing them at multiples of discrimination threshold contrast. In a subsequent electroencephalographic experiment observers were asked to classify contours as objects or non-objects. An advantage in accuracy was found for full-color stimuli over the reduced-color stimuli but only if the contours depicted objects as opposed to non-objects. Event-related potentials revealed the neural correlate of this object-specific luminance advantage. The amplitude of the centro-occipital N1 component was modulated by stimulus class with the effect being driven by the presence of luminance information. We conclude that high-level discrimination processes in the cortex start relatively early and exhibit object-selective effects only in the presence of luminance information. This is consistent with the superiority of luminance in subserving object identification processes. PMID- 21632773 TI - Preparation of mouse embryos for optical projection tomography imaging. PMID- 21632771 TI - Herpes simplex virus immediate-early protein ICP0 is targeted by SIAH-1 for proteasomal degradation. AB - Herpes simplex virus (HSV) immediate-early protein ICP0 is a transcriptional activator with E3 ubiquitin ligase activity that induces the degradation of ND10 proteins, including the promyelocytic leukemia protein (PML) and Sp100. Moreover, ICP0 has a role in the derepression of viral genomes and in the modulation of the host interferon response to virus infection. Here, we report that ICP0 interacts with SIAH-1, a cellular E3 ubiquitin ligase that is involved in multiple cellular pathways and is itself capable of mediating PML degradation. This novel virus host interaction profoundly stabilized SIAH-1 and recruited this cellular E3 ligase into ICP0-containing nuclear bodies. Moreover, SIAH-1 mediated the polyubiquitination of HSV ICP0 in vitro and in vivo. After infection of SIAH-1 knockdown cells with HSV, higher levels of ICP0 were produced, ICP0 was less ubiquitinated, and the half-life of this multifunctional viral regulatory protein was increased. These results indicate an inhibitory role of SIAH-1 during lytic infection by targeting ICP0 for proteasomal degradation. PMID- 21632774 TI - IMGT/3Dstructure-DB: querying the IMGT database for 3D structures in immunology and immunoinformatics (IG or antibodies, TR, MH, RPI, and FPIA). PMID- 21632775 TI - IMGT/DomainGapAlign: IMGT standardized analysis of amino acid sequences of variable, constant, and groove domains (IG, TR, MH, IgSF, MhSF). PMID- 21632776 TI - IMGT/Collier de Perles: IMGT standardized representation of domains (IG, TR, and IgSF variable and constant domains, MH and MhSF groove domains). PMID- 21632777 TI - IMGT/junctionanalysis: IMGT standardized analysis of the V-J and V-D-J junctions of the rearranged immunoglobulins (IG) and T cell receptors (TR). PMID- 21632778 TI - IMGT/V-QUEST: IMGT standardized analysis of the immunoglobulin (IG) and T cell receptor (TR) nucleotide sequences. PMID- 21632779 TI - Dissection and staining of Drosophila optic lobes at different stages of development. PMID- 21632780 TI - Introducing recombinant picornaviral genomes into cells. PMID- 21632781 TI - Isoelectric focusing of iTRAQ-labeled yeast. PMID- 21632782 TI - iTRAQ-labeled yeast peptide clean-up using a reversed-phase column. PMID- 21632783 TI - Labeling yeast peptides with the iTRAQ reagent. PMID- 21632784 TI - Preparation of peptides from yeast cells for iTRAQ analysis. PMID- 21632785 TI - Optical projection tomography of vertebrate embryo development. PMID- 21632786 TI - IMGT, the International ImMunoGeneTics Information System. PMID- 21632787 TI - Engineering light-regulated ion channels. PMID- 21632788 TI - IMGT Collier de Perles for the variable (V), constant (C), and groove (G) domains of IG, TR, MH, IgSF, and MhSF. PMID- 21632789 TI - IMGT unique numbering for the variable (V), constant (C), and groove (G) domains of IG, TR, MH, IgSF, and MhSF. PMID- 21632790 TI - From IMGT-ONTOLOGY CLASSIFICATION Axiom to IMGT standardized gene and allele nomenclature: for immunoglobulins (IG) and T cell receptors (TR). PMID- 21632791 TI - From IMGT-ONTOLOGY DESCRIPTION axiom to IMGT standardized labels: for immunoglobulin (IG) and T cell receptor (TR) sequences and structures. PMID- 21632792 TI - From IMGT-ONTOLOGY IDENTIFICATION axiom to IMGT standardized keywords: for immunoglobulins (IG), T cell receptors (TR), and conventional genes. PMID- 21632793 TI - The milky (path)way of proteinosis: has the time come. PMID- 21632794 TI - Tuberculosis: an ancient and evergreen disease. PMID- 21632795 TI - Update on tuberculosis: TB in the early 21st century. PMID- 21632796 TI - Antifibrotic activities of pirfenidone in animal models. AB - Pirfenidone is an orally active small molecule that has recently been evaluated in large clinical trials for the treatment of idiopathic pulmonary fibrosis, a fatal disease in which the uncontrolled deposition of extracellular matrix leads to progressive loss of lung function. This review describes the activity of pirfenidone in several well-characterised animal models of fibrosis in the lung, liver, heart and kidney. In these studies, treatment-related reductions in fibrosis are associated with modulation of cytokines and growth factors, with the most commonly reported effect being reduction of transforming growth factor-beta. The consistent antifibrotic activity of pirfenidone in a broad array of animal models provides a strong preclinical rationale for the clinical characterisation of pirfenidone in pulmonary fibrosis and, potentially, other conditions with a significant fibrotic component. PMID- 21632797 TI - Pulmonary alveolar proteinosis. AB - Pulmonary alveolar proteinosis (PAP) is a rare pulmonary disease characterised by alveolar accumulation of surfactant. It may result from mutations in surfactant proteins or granulocyte macrophage-colony stimulating factor (GM-CSF) receptor genes, it may be secondary to toxic inhalation or haematological disorders, or it may be auto-immune, with anti-GM-CSF antibodies blocking activation of alveolar macrophages. Auto-immune alveolar proteinosis is the most frequent form of PAP, representing 90% of cases. Although not specific, high-resolution computed tomography shows a characteristic "crazy paving" pattern. In most cases, bronchoalveolar lavage findings establish the diagnosis. Whole lung lavage is the most effective therapy, especially for auto-immune disease. Novel therapies targeting alveolar macrophages (recombinant GM-CSF therapy) or anti-GM-CSF antibodies (rituximab and plasmapheresis) are being investigated. Our knowledge of the pathophysiology of PAP has improved in the past 20 yrs, but therapy for PAP still needs improvement. PMID- 21632798 TI - Towards a better diagnosis of idiopathic pulmonary fibrosis. AB - Idiopathic pulmonary fibrosis (IPF) is the most common of the idiopathic interstitial pneumonias, and poses significant clinical challenges. IPF diagnosis is based on clear-cut computed tomography (CT) and histopathological criteria, in an appropriate clinical context. The diagnostic criteria include: 1) exclusion of known causes of interstitial lung disease (including connective tissue disease); 2) usual interstitial pneumonia pattern on high-resolution CT in patients not subjected to surgical lung biopsy; and 3) specific combinations of high resolution CT with pathological patterns in case of surgical lung biopsy. Improved diagnosis of IPF may help physicians to reduce the delay before an accurate diagnosis is made and increase patient awareness and access to adequate information, follow-up and treatment. PMID- 21632799 TI - Follow-up and nonpharmacological management of the idiopathic pulmonary fibrosis patient. AB - Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, fatal form of diffuse interstitial lung disease. Management of IPF requires an orderly approach, with regular evaluations and implementation of both pharmacological and nonpharmacological treatments. Pulmonary rehabilitation can relieve patients from the distressing symptoms of IPF and improve quality of life. Oxygen therapy is central to treatment of all patients. Lung transplantation enhances survival in selected patients. Mechanical ventilation may be used in patients with acute exacerbations, but the prognosis is poor in these cases. Palliative care focuses on symptom management, advance directives and end-of-life planning. Patient support groups may also play an important role. PMID- 21632800 TI - Iatrogenic eosinophilic pleural effusion. PMID- 21632801 TI - Blind needle biopsy of the pleura: why not? PMID- 21632805 TI - Rare metastases of differentiated thyroid carcinoma: pictorial review. AB - Differentiated thyroid cancer (DTC) is usually indolent with good prognosis and long-term survival. However, DTC distant metastasis is often a grave event and accounts for most of its disease-specific mortality. The major sites of distant metastases are the lung and bone. Metastases to the brain, breast, liver, kidney, muscle, and skin are rare or relatively rare. Nevertheless, recognizing rare metastases from DTC has a significant impact on the clinical decision making and prognosis of patients. (131)I single photon emission computed tomography/computed tomography ((131)I-SPECT/CT) can provide both metabolic and anatomic information about a lesion; therefore, it can better localize and define the (131)I-WBS findings in DTC patients. In this pictorial review, the imaging features of a range of rare metastases from DTC are demonstrated, with a particular emphasis on the (131)I-SPECT/CT diagnostic aspect. PMID- 21632806 TI - Hooked on zebrafish: insights into development and cancer of endocrine tissues. AB - Zebrafish is emerging as a unique model organism for studying cancer genetics and biology. For several decades zebrafish have been used to study vertebrate development, where they have made important contributions to understanding the specification and differentiation programs in many tissues. Recently, zebrafish studies have led to important insights into thyroid development, and have been used to model endocrine cancer. Zebrafish possess a unique set of attributes that make them amenable to forward and reverse genetic approaches. Zebrafish embryos develop rapidly and can be used to study specific cell lineages or the effects of chemicals on pathways or tissue development. In this review, we highlight the structure and function of endocrine organs in zebrafish and outline the major achievements in modeling cancer. Our goal is to familiarize readers with the zebrafish as a genetic model system and propose opportunities for endocrine cancer research in zebrafish. PMID- 21632807 TI - Kisspeptin-10 is a potent stimulator of LH and increases pulse frequency in men. AB - CONTEXT: Kisspeptins stimulate GnRH and thus gonadotropin secretion. Kisspeptin 10 is the minimal kisspeptin sequence with full intrinsic bioactivity, but it has not been studied in man. OBJECTIVE: We investigated our hypothesis that kisspeptin-10 increases GnRH and thus LH pulse frequency. DESIGN AND PARTICIPANTS: The dose response of kisspeptin-10 was investigated by administering iv bolus doses (0.01-3.0 MUg/kg) and vehicle to healthy men. Effects on LH pulse frequency and size were determined by deconvolution analysis during infusion of kisspeptin-10 for up to 22.5 h. RESULTS: Intravenous bolus kisspeptin-10 resulted in a rapid and dose-dependent rise in serum LH concentration, with maximal stimulation at 1 MUg/kg (4.1 +/- 0.4 to 12.4 +/- 1.7 IU/liter at 30 min, P < 0.001, n = 6). Administration of 3 MUg/kg elicited a reduced response vs. 1 MUg/kg (P < 0.05). Infusion of kisspeptin-10 at 4 MUg/kg . h for 22.5 h elicited an increase in LH from a mean of 5.4 +/- 0.7 to 20.8 +/- 4.9 IU/liter (n = 4; P < 0.05) and serum testosterone increased from 16.6 +/- 2.4 to 24.0 +/- 2.5 nmol/liter (P < 0.001). LH pulses were obscured at this high rate of secretion, but a lower dose infusion of kisspeptin-10 (1.5 MUg/kg . h) increased mean LH from 5.2 +/- 0.8 to 14.1 +/- 1.7 IU/liter (n = 4; P < 0.01) and increased LH pulse frequency from 0.7 +/- 0.1 to 1.0 +/- 0.2 pulses/h (P < 0.05) and secretory burst mass from 3.9 +/- 0.4 to 12.8 +/- 2.6 IU/liter (P < 0.05). CONCLUSIONS: Kisspeptin-10 boluses potently evoke LH secretion in men, and continuous infusion increases testosterone, LH pulse frequency, and pulse size. Kisspeptin analogues have therapeutic potential as regulators of LH and thus testosterone secretion. PMID- 21632808 TI - Cotreatment with pegvisomant and a somatostatin analog (SA) in SA-responsive acromegalic patients. AB - CONTEXT: Cotreatment of acromegaly with pegvisomant and a somatostatin analog (SA) has proven feasible. Previous studies in the field have focused on patients with an insufficient response to SA monotherapy in whom pegvisomant was added without changing the SA dose. OBJECTIVE: The objective of the study was to study whether patients sufficiently controlled on SA monotherapy can be transferred to combination therapy with low-dose pegvisomant and a reduced SA dose. DESIGN: Eighteen acromegalic patients well controlled on SA monotherapy, mean +/- se aged 54 +/- 3 yr, were randomized in a parallel study over 24 wk to unchanged SA monotherapy or cotreatment with pegvisomant (15-30 mg twice a week) and SA (half the usual dosage). SETTING: This was an investigator-initiated study in a single tertiary referral center. MAIN OUTCOME MEASURES: Glucose tolerance, substrate metabolism, insulin sensitivity, body composition, and quality of life were measured. RESULTS: Median pegvisomant dose was 52.5 mg/wk (range 30-60). IGF-I (micrograms per liter) was comparable both at baseline (P = 0.88) and after 24 wk of treatment (P = 0.48). The change in IGF-I between baseline and wk 24 also did not differ between groups (P = 0.15). Apart from increased peak insulin levels during the oral glucose tolerance test in the cotreatment group, no substantial differences between the two groups were detected. Moderately elevated liver enzymes were found in 17% of the patients on pegvisomant therapy. CONCLUSION: Acromegalic patients well controlled on SA monotherapy can maintain safe IGF-I levels during 24 wk of cotreatment with low-dose pegvisomant and a 50% reduced SA dose. This treatment modality, however, does not seem to provide significant benefits for the patients. PMID- 21632809 TI - Acute peripheral metabolic effects of intraarterial leg infusion of somatostatin in healthy young men. AB - CONTEXT: Evidence suggests that somatostatin not only inhibits the secretion of GH but also suppresses GH action in peripheral tissues. OBJECTIVE: We tested the hypothesis that somatostatin suppresses GH activity in human skeletal muscle in vivo. DESIGN AND PARTICIPANTS: Eight healthy young men (25.3 +/- 2.8 yr) were studied on a single occasion after an overnight fast for 4 h [including a basal period (0-2 h) and a hyperinsulinemic euglycemic clamp (2-4 h)] during an iv GH infusion (50 ng/kg-1 . min-1). Each subject received an intraarterial somatostatin infusion (150 MUg/h-1) into one femoral artery and an intraarterial saline infusion into the contra lateral artery. The simultaneous blood samples were drawn from both femoral veins. Muscle biopsies were obtained from one leg at t = 0 and from both legs during the basal period and during the clamp. MAIN OUTCOME MEASURES: Muscle glucose uptake, signaling proteins for GH (phosphorylated signal transducer and activator of transcription-5) and insulin (phosphorylation of AS160), and expression of GH-regulated genes (IGF-I and suppressor of cytokine signaling 1-3) were measured. RESULTS: Somatostatin significantly increased glucose uptake measured by arteriovenous glucose difference during the basal period (P = 0.03) but not during the clamp. There was a tendency for the phosphorylation of AS160 to be higher in the somatostatin infused leg compared with the saline leg (P = 0.055). The expression of suppressor of cytokine signaling-1 mRNA was significantly elevated in the clamp biopsy from the saline-infused leg (P = 0.024). CONCLUSIONS: We concluded the following: 1) in the presence of systemic GH exposure, somatostatin increases basal glucose uptake and reduces the expression of GH-regulated genes directly in skeletal muscle; 2) this supports the concept that somatostatin suppresses GH activity in peripheral tissues, and 3) this may add to the therapeutic effects of somataostatin analogs. PMID- 21632810 TI - Bioavailability of vitamin D in malnourished adolescents with anorexia nervosa. AB - CONTEXT: Young women with anorexia nervosa (AN) have a normal vitamin D status. The bioavailability of vitamin D during malnutrition is unknown. OBJECTIVE: The objective of the study was to examine the serum response to oral ergocalciferol in AN. DESIGN/SETTING: This was a prospective cohort study, conducted in 2007 2009 at a tertiary care center. PATIENTS/INTERVENTIONS: Twelve adolescents with AN (age 19.6 +/- 2.0 yr, body mass index 16.5 +/- 1.4 kg/m2) and 12 matched healthy controls (20.0 +/- 2.4 yr, 22.7 +/- 1.0 kg/m2) received one baseline 50,000 IU oral dose of ergocalciferol. MAIN OUTCOMES: Serum D2, D3, 25 hydroxyvitamin D, and 1,25-dihydroxyvitamin D, collected before ingestion, at 6 and 24 h and weekly for 4 wk, and body composition measures were measured. RESULTS: The AN group was severely malnourished (77.2 +/- 6.3% median body weight), whereas the control group was normal weighted (106.2 +/- 6.2%). From a common baseline D2 (1.5 +/- 1.6 nmol/liter, P =0.34) the groups diverged (time * group interaction P = 0.04), peaking at 70 +/- 34 nmol/liter at 6 h in controls compared with 43 +/- 28 nmol/liter in AN subjects (P = 0.008). The D2 trajectories converged at 24 h (57 nmol/liter, P = 0.98) and returned to near baseline at 1 wk. Baseline D3 was higher in AN subjects (12.1 +/- 9.6 vs. 3.1 +/- 2.3 nmol/liter, P < 0.001) and remained higher throughout. 25-Hydroxyvitamin D followed a common trajectory (time * group interaction P = 0.15), rising to 45 +/ 10 nmol/liter at 24 h but returning to baseline by wk 3 (P = 0.36). Correlating vitamin D levels with fat measures (body mass index, body fat) produced similar findings. CONCLUSIONS: Despite severe malnutrition, young women with AN had a similar bioavailability of oral ergocalciferol as the healthy-weighted controls. Vitamin D dosing for patients suffering from malnutrition may not differ from that for normal-weighted adolescents. PMID- 21632811 TI - Early metformin therapy (age 8-12 years) in girls with precocious pubarche to reduce hirsutism, androgen excess, and oligomenorrhea in adolescence. AB - CONTEXT: Girls with a combined history of low(-normal) birth weight (LBW) and precocious pubarche (PP) are at high risk to develop polycystic ovary syndrome (PCOS). OBJECTIVE: The objective of the study was to compare the capacity of early vs. late metformin treatment to prevent adolescent PCOS. DESIGN: This was a randomized, open-label study over 7 yr. SETTING: The study was conducted at a university hospital. PATIENTS: Thirty-eight LBW-PP girls were followed up from the mean age 8 until age 15 yr. INTERVENTION: Early metformin (study yr 1-4; age 8-12 yr) vs. late metformin (yr 6; age 13-14 yr). MAIN OUTCOME MEASURES: Measures included height; weight; hirsutism score; menstrual cycle; endocrine-metabolic screening (fasting; follicular phase); C-reactive protein; body composition (absorptiometry); abdominal fat partitioning (magnetic resonance imaging); ovarian morphology (ultrasound); PCOS (National Institutes of Health and Androgen Excess Society definitions) after yr 7 (all girls thus untreated for at least 1 yr). RESULTS: None of the girls dropped out of the study. At age 15 yr, early metformin girls were taller (4 cm), were in a less proinflammatory state, and had less central fat due to reductions in visceral and hepatic fat. Hirsutism, androgen excess, oligomenorrhea, and PCOS were between 2- and 8-fold more prevalent in late- than early-treated girls. Abdominal adiposity was the first variable to diverge (at age 8-10 yr) between girls without vs. with PCOS at age 15 yr. CONCLUSIONS: In LBW-PP girls, early metformin therapy was found to prevent or delay the development of hirsutism, androgen excess, oligomenorrhea, and PCOS more effectively than late metformin. The time window of late childhood and early puberty may be more critical for the development, and thus for the prevention, of adolescent PCOS than the first years beyond menarche. PMID- 21632812 TI - Is the incidence of congenital hypothyroidism really increasing? A 20-year retrospective population-based study in Quebec. AB - CONTEXT: Congenital hypothyroidism (CH) is reportedly increasing in the United States, possibly reflecting changes in screening methods. In Quebec, the same initial TSH cutoff (15 mU/liter) has been used for the last 20 yr, but in 2001, the cutoff was decreased from 15 to 5 mU/liter for the second test, which is requested when TSH is intermediate (15-30 mU/liter) on the first. OBJECTIVES: Our objective was to assess the incidence of CH over the last 20 yr in Quebec. DESIGN, SETTING, PATIENTS, AND MAIN OUTCOME MEASURE: This is a population-based retrospective study. Incidences by etiology based on thyroid scintigraphy with technetium were compared between 1990-2000 and 2001-2009. RESULTS: Of 1,660,857 newborns over 20 yr, 620 had CH (incidence 1:2679). Etiology was dysgenesis (n = 389, 1:4270), either ectopy (n = 290) or athyreosis (n = 99), goiter (n = 52, 1:31,940), normal-size gland in situ (n = 115, 1:14,442), and unknown (n = 64, 1:25,950). The new screening algorithm identified 49 additional cases (i.e. 25 normal-size gland in situ, 12 unknown etiology, 10 ectopies, and two goiters). Consequently, the incidence of normal-size gland in situ or of unknown etiology more than doubled (1:22,222 to 1:9,836, P = 0.0015; and 1:43,824 to 1:17,143, P = 0.0018, respectively) but that of dysgenesis and goiter remained stable. Had the 1990-2000 algorithm been applied in 2001-2009, no change in incidence would have been observed in any category. CONCLUSION: Estimating the incidence of CH is influenced by minimal changes in TSH screening cutoffs. Lower cutoffs identify additional cases that have predominantly functional disorders whose impact on intellectual disability, if left untreated, remains to be determined. PMID- 21632813 TI - Medical and surgical evaluation and treatment of adrenal incidentalomas. AB - INTRODUCTION: Adrenal incidentalomas are detected in approximately 4% of patients undergoing high-resolution abdominal imaging studies. The majority of adrenal incidentalomas are benign, but careful evaluation of all patients is warranted to be certain that primary adrenocortical carcinoma and functional adenomas are not missed. METHODS: The diagnostic approach in patients with adrenal incidentalomas should focus on two main questions: whether the lesion is malignant, and whether it is hormonally active. Radiological evaluation including noncontrast and contrast computed tomography attenuation values expressed in Hounsfield units is the best tool to differentiate between benign and malignant adrenal masses. All adrenal tumors with suspicious radiological findings, most functional tumors, and all tumors more than 4 cm in size that lack characteristic benign imaging features should be surgically excised. All patients should undergo hormonal evaluation for subclinical Cushing's syndrome and pheochromocytoma, and those with hypertension should also be evaluated for hyperaldosteronism. Combined 1-mg dexamethasone suppression test, plasma metanephrines, and aldosterone/plasma renin activity measurements (if hypertensive) are reasonable initial hormonal evaluations. RESULTS: Annual biochemical follow-up of most patients with adrenal incidentalomas, especially if the tumor is more than 3 cm in size, for up to 5 yr may be reasonable. Patients with adrenal masses less than 4 cm in size and a noncontrast attenuation value of more than 10 Hounsfield units should have a repeat computed tomography study in 3-6 months and then yearly for 2 yr. Adrenal tumors with indeterminate radiological features that grow to at least 0.8 cm over 3-12 months may be considered for surgical resection. PMID- 21632814 TI - Adiponectin and lipid profiles compared with insulins in relation to early growth of British South Asian and European children: the Manchester children's growth and vascular health study. AB - CONTEXT: Adiponectin, high-density lipoprotein cholesterol (HDL-C) and insulin concentrations may be important in the pathophysiology of cardiovascular disease. OBJECTIVE: We tested the hypothesis that serum adiponectin rather than insulin differs from early life, between South Asians and Europeans, with a potentially key role in excess cardiovascular risk characteristic of adult South Asians. DESIGN AND PARTICIPANTS: We conducted a longitudinal study of 215 British-born children of European (n = 138) and South Asian (n = 77) origin, from birth to 3 yr. MAIN OUTCOME MEASURE: Serum adiponectin, insulin, proinsulin and HDL-C concentrations were assessed in relation to ethnic group and growth in anthropometric variables from 0-3 yr of age. RESULTS: Serum adiponectin was lower in South Asian children, despite their smaller size, notable at age 3-6 months (9.5 vs. 11.8 mg/liter; P = 0.04), with no ethnic differences in serum lipids or insulin or proinsulin. In mixed-effects longitudinal models for HDL-C, determinants were adiponectin (P = 0.034), age (P < 0.001), and body mass index (P < 0.001) but not ethnicity. None of these or growth variables affected either insulin or proinsulin. In a fully adjusted mixed-effects longitudinal model including age, sex, insulin, and proinsulin, the independent determinants of serum adiponectin were height [21.3 (95% confidence interval = 31.7-10.8 cm lower, for every 1 mmol/liter increase in adiponectin, P < 0.001], HDL-C [2.8 (1.3-4.2) mmol/liter higher, P < 0.0001], body mass index (lower, P = 0.03), and South Asian ethnicity (lower, P = 0.01). CONCLUSIONS: These British South Asian origin infants have lower serum adiponectin but no differences in HDL-C or insulin molecules. In South Asians, factors affecting adiponectin metabolism in early life, rather than insulin resistance, likely determine later excess cardiovascular risk. PMID- 21632815 TI - Effects of a single dose of exenatide on appetite, gut hormones, and glucose homeostasis in adults with Prader-Willi syndrome. AB - CONTEXT: Prader-Willi syndrome (PWS) is associated with hyperphagia and obesity, without effective pharmacological treatment. Exenatide, recently developed for treatment of type 2 diabetes, induces appetite suppression and weight loss with common side effects. OBJECTIVE: The objective of the study was to investigate the initial safety and effectiveness of exenatide in adult PWS subjects compared with obese controls (OBESE). DESIGN, SETTING, PATIENTS, AND INTERVENTION: Eight PWS and 11 OBESE patients underwent standardized meal studies after a single sc injection of 10 MUg exenatide or placebo in a single-blinded, crossover design. MAIN OUTCOME MEASURES: Glucose, insulin, C-peptide, glucagon, peptide YY (PYY; total)/PYY (3-36), glucagon-like peptide-1, and ghrelin (total) were measured fasting and postprandially. Appetite and satiety were assessed by visual analog scales. Energy expenditure (EE) was measured by indirect calorimetry. Side effects were screened during and for 24 h after the meal. RESULTS: PWS and OBESE patients were matched for gender, age, body mass index, and central/total body fat. In both groups, exenatide increased satiety and lowered glucose and insulin levels but increased insulin secretion rate. Side effects were absent in PWS but common in OBESE patients. During the meal, PYY (total) and ghrelin were elevated in PWS patients. Exenatide decreased PYY (total) and glucagon-like peptide-1, whereas ghrelin remained unchanged. Energy expenditure was unchanged by exenatide. CONCLUSIONS: Our pilot study demonstrates that exenatide is well tolerated in PWS patients. It increases satiety independently of measured appetite hormones, exerting glucose lowering, and insulinotropic effects similarly in PWS and OBESE patients. Larger prospective studies should investigate whether chronic exenatide administration will reduce hyperphagia and overweight in PWS patients without side effects. PMID- 21632816 TI - Effect of intensive insulin therapy on the somatotropic axis of critically ill children. AB - CONTEXT: Intensive insulin therapy (IIT) improved outcome in the adult and pediatric intensive care unit (PICU) compared with conventional insulin therapy (CIT). IIT did not increase the anabolic hormone IGF-I in critically ill adults, but feeding in critically ill children and pediatric hormonal responses may differ. Twenty-five percent of the children with IIT experienced hypoglycemia, which may have evoked counterregulatory responses. OBJECTIVE: We hypothesized that IIT reactivates the somatotropic axis and anabolism in PICU patients. DESIGN: This was a preplanned subanalysis of a randomized controlled trial on IIT. PATIENTS: We studied 369 patients who stayed in PICU for at least 3 d (study 1) and 126 patients in a nested case-control study (study 2). MAIN OUTCOME MEASURES: Circulating insulin, C-peptide, GH, IGF-I, bioavailable IGF-I, IGF binding protein (IGFBP)-1, IGFBP-3, and acid-labile subunit were analyzed upon admission and d 3. In the nested case-control study, the somatotropic axis, cortisol, and glucagon were analyzed before and after hypoglycemia. RESULTS: On d 3, C-peptide was more than 10-fold lower (P < 0.0001) in the IIT group than in the CIT group. IIT increased circulating GH (P = 0.04) and lowered bioavailable IGF-I (P = 0.002). IIT also decreased IGFBP-3 (P = 0.0005) and acid-labile subunit (P = 0.007), while increasing IGFBP-1 (P = 0.04) and the urea/creatinine ratio, a marker of catabolism (P = 0.03). In the nested case-control study, IGFBP 1 was increased after hypoglycemia, whereas the somatotropic axis and the counterregulatory hormones cortisol and glucagon did not change. CONCLUSIONS: Despite improved PICU outcome, IIT did not counteract the catabolic state of critical illness. Suppression of portal insulin may have resulted in lower bioavailable IGF-I. PMID- 21632817 TI - Cardiac dimensions are largely determined by dietary salt in patients with primary aldosteronism: results of a case-control study. AB - CONTEXT: Animal studies have demonstrated that dietary sodium intake is a major influence in the pathogenesis of aldosterone-induced effects in the heart such as left ventricular (LV) hypertrophy and fibrosis. LV hypertrophy is an important predictor for cardiovascular morbidity and mortality. OBJECTIVE: We aimed to investigate the relationships between aldosterone and dietary salt and LV dimensions in patients with primary aldosteronism (PA). DESIGN AND PARTICIPANTS: This case-control study included 21 patients with confirmed PA and 21 control patients with essential hypertension matched for age, gender, duration of hypertension, and 24-h systolic and diastolic blood pressure. MAIN OUTCOME MEASURES: Patients were evaluated by echocardiography and 24-h urinary sodium (UNa) excretion while consuming their usual diets. RESULTS: Patients with PA had significantly greater mean LV end-diastolic diameter, interventricular septum and posterior wall thicknesses, LV mass (LVM) and LV mass index, and end systolic and diastolic volumes than control patients. UNa significantly positively correlated with interventricular septum, posterior wall thicknesses, and LVM in the patients with PA but not in control patients. In a multivariate analysis, UNa was an independent predictor for LV wall thickness and LV mass among the patients with PA but not in patients with essential hypertension. CONCLUSIONS: These findings emphasize the importance of dietary sodium in determining the degree of cardiac damage in those patients with PA, and we suggest that aldosterone excess may play a permissive role. In patients with PA, because a high-salt diet is associated with greater LVM, dietary salt restriction might reduce cardiovascular risk. PMID- 21632818 TI - Growth differentiating factor 9 (GDF9) and bone morphogenetic protein 15 both activate development of human primordial follicles in vitro, with seemingly more beneficial effects of GDF9. AB - CONTEXT: The signals initiating growth of primordial follicles are unknown. Bone morphogenetic protein 15 (BMP15) and growth differentiating factor 9 (GDF9) are promising candidates. OBJECTIVE: The objective of the study was to evaluate for the first time the effects of human recombinant BMP15 and human recombinant GDF9 on the in vitro development of human primordial follicles. DESIGN AND SETTING: This was a controlled culture study performed in a major tertiary university affiliated medical center. MATERIALS: Materials included ovarian tissue from 17 girls/women and three aborted human fetuses. INTERVENTION: There were no interventions. MAIN OUTCOME MEASURE: Histological and immunohistochemical (proliferating cell nuclear antigen, BMP15, and GDF9) studies and an endocrine assay of 17beta-estradiol were conducted. RESULTS: In the samples from girls/women, the number of developing follicles was greater with GDF9 or BMP15 alone than with no BMP15 or GDF9. Higher 17beta-estradiol secretion was noted after treatment with GDF9 than with BMP15 or with GDF9+anti-GDF9. The number of atretic follicles was greater with BMP15 than with GDF9. Proliferating cell nuclear antigen expression was greater with the higher dose of both growth factors than the lower dose. Expression of BMP15 and GDF9 was identified in samples cultured without BMP15 or GDF9. Results for the fetal follicles yielded no distinguishable pattern. CONCLUSIONS: Although both BMP15 and GDF9 promoted activation of human primordial follicles from girls/women (but not human fetuses) in a dose-dependent manner, GDF9 seems more beneficial. PMID- 21632819 TI - In nonfunctional pituitary adenomas, estrogen receptors and slug contribute to development of invasiveness. AB - CONTEXT: Clinical studies suggest that an imbalance in the actions of estrogen receptor (ER)-alpha and ERbeta is associated with invasion of tumors of epithelial origin. Both ER have been detected in the pituitary adenomas (PA). Our previous study together with other reports suggests that an imbalance between ERalpha and ERbeta contributes to the pathogenesis and biological behavior of PA including invasion. However, the roles of the two ER in invasiveness of PA have not been clarified. OBJECTIVE: The expression of ER, Slug, and E-cadherin in 41 cases of nonfunctional PA (NFPA) were determined to evaluate whether ER were related to the invasiveness of NFPA. Furthermore, we aimed by analysis of the correlation between ER and E-cadherin or Slug to understand molecular mechanisms related to invasiveness. METHODS: Immunohistochemistry, RT-PCR, and Western blot were performed. RESULTS: Nuclear ERalpha staining was significantly stronger in invasive NFPA than noninvasive ones (P < 0.01). In contrast, nuclear ERbeta staining was significantly weaker in invasive NFPA than in noninvasive ones (P < 0.01). Both E-cadherin mRNA and protein were decreased in invasive NFPA compared with noninvasive ones. Moreover, Slug, a repressor of E-cadherin, was significantly increased in invasive over noninvasive NFPA (P < 0.01). There were significant correlations between ER and Slug or E-cadherin in NFPA, in which Slug was positively correlated with ERalpha and inversely correlated with ERbeta, whereas E-cadherin was positively correlated with ERbeta and inversely correlated with ERalpha. CONCLUSIONS: ERalpha and ERbeta may act in opposite directions to regulate the Slug-E-cadherin pathway and to affect invasiveness of NFPA. PMID- 21632820 TI - RNAi in Xenopus: look before you leap. AB - RNAi has revolutionized reverse genetics; however, RNAi is not necessarily ubiquitous or constitutive. Lund and colleagues (pp. 1121-1131) show that microRNA (miRNA) effector Argonautes (Agos) are limiting and easily saturated during early Xenopus embryogenesis. Moreover, this stage is devoid of slicing capacity. Supplementation of Ago proteins rescued endogenous miRNA activity in the presence of exogenous siRNAs, and, excitingly, ectopic Ago2 could now support RNAi in Xenopus. These observations may potentially facilitate RNAi in other recalcitrant model organisms. PMID- 21632821 TI - Regulation of the Dbp5 ATPase cycle in mRNP remodeling at the nuclear pore: a lively new paradigm for DEAD-box proteins. AB - It is commonly assumed that all DEAD-box ATPases function via a shared mechanism, since this is the case for the few proteins characterized thus far. Hodge and colleagues (pp. 1052-1064) and Noble and colleagues (pp. 1065-1077) now describe a novel model for Dbp5's ATPase cycle in mRNA (messenger RNA)/protein complex (mRNP) remodeling during nuclear export. Notably, unlike other DEAD-box proteins, Dbp5 uses a conformational change distinct from ATP hydrolysis for its activity and requires an ADP release factor to reset its ATPase cycle. PMID- 21632822 TI - Different types of oscillations in Notch and Fgf signaling regulate the spatiotemporal periodicity of somitogenesis. AB - Somitogenesis is controlled by cyclic genes such as Notch effectors and by the wave front established by morphogens such as Fgf8, but the precise mechanism of how these factors are coordinated remains to be determined. Here, we show that effectors of Notch and Fgf pathways oscillate in different dynamics and that oscillations in Notch signaling generate alternating phase shift, thereby periodically segregating a group of synchronized cells, whereas oscillations in Fgf signaling released these synchronized cells for somitogenesis at the same time. These results suggest that Notch oscillators define the prospective somite region, while Fgf oscillators regulate the pace of segmentation. PMID- 21632823 TI - Methylation specifies distinct estrogen-induced binding site repertoires of CBP to chromatin. AB - Multiple signaling pathways ultimately modulate the epigenetic information embedded in the chromatin of gene promoters by recruiting epigenetic enzymes. We found that, in estrogen-regulated gene programming, the acetyltransferase CREB binding protein (CBP) is specifically and exclusively methylated by the coactivator-associated arginine methyltransferase (CARM1) in vivo. CARM1 dependent CBP methylation and p160 coactivators were required for estrogen induced recruitment to chromatin targets. Notably, methylation increased the histone acetyltransferase (HAT) activity of CBP and stimulated its autoacetylation. Comparative genome-wide chromatin immunoprecipitation sequencing (ChIP-seq) studies revealed a variety of patterns by which p160, CBP, and methyl CBP (meCBP) are recruited (or not) by estrogen to chromatin targets. Moreover, significant target gene-specific variation in the recruitment of (1) the p160 RAC3 protein, (2) the fraction of a given meCBP species within the total CBP, and (3) the relative recruitment of different meCBP species suggests the existence of a target gene-specific "fingerprint" for coregulator recruitment. Crossing ChIP seq and transcriptomics profiles revealed the existence of meCBP "hubs" within the network of estrogen-regulated genes. Together, our data provide evidence for an unprecedented mechanism by which CARM1-dependent CBP methylation results in gene-selective association of estrogen-recruited meCBP species with different HAT activities and specifies distinct target gene hubs, thus diversifying estrogen receptor programming. PMID- 21632824 TI - The yeast rapid tRNA decay pathway primarily monitors the structural integrity of the acceptor and T-stems of mature tRNA. AB - tRNAs, like other RNAs, are subject to quality control steps during and after biosynthesis. We previously described a rapid tRNA degradation (RTD) pathway in which the 5'-3' exonucleases Rat1 and Xrn1 degrade mature tRNA(Val(AAC)) in yeast mutants lacking m(7)G and m(5)C, and mature tRNA(Ser(CGA)) in mutants lacking Um and ac(4)C. To understand how the RTD pathway selects substrate tRNAs among different tRNAs lacking the same modifications, we used a genetic screen to examine tRNA(Ser(CGA)) variants. Our results suggest that RTD substrate recognition in vivo depends primarily on the stability of the acceptor and T stems, and not the anti-codon stem, and does not necessarily depend on modifications, since fully modified tRNAs are subject to RTD if appropriately destabilized. We found that weaker predicted stability of the acceptor and T stems of tRNAs is strongly correlated with RTD sensitivity, increased RNase T2 sensitivity of this region of the tRNA in vitro, and increased exposure of the 5' end to phosphatase. We also found that purified Xrn1 selectively degrades RTD substrate tRNAs in vitro under conditions in which nonsubstrates are immune. These results suggest that tRNAs have evolved not only for accurate translation, but for resistance to attack by RTD. PMID- 21632825 TI - Foxl1-Cre-marked adult hepatic progenitors have clonogenic and bilineage differentiation potential. AB - Isolation of hepatic progenitor cells is a promising approach for cell replacement therapy of chronic liver disease. The winged helix transcription factor Foxl1 is a marker for progenitor cells and their descendants in the mouse liver in vivo. Here, we purify progenitor cells from Foxl1-Cre; RosaYFP mice and evaluate their proliferative and differentiation potential in vitro. Treatment of Foxl1-Cre; RosaYFP mice with a 3,5-diethoxycarbonyl-1,4-dihydrocollidine diet led to an increase of the percentage of YFP-labeled Foxl1(+) cells. Clonogenic assays demonstrated that up to 3.6% of Foxl1(+) cells had proliferative potential. Foxl1(+) cells differentiated into cholangiocytes and hepatocytes in vitro, depending on the culture condition employed. Microarray analyses indicated that Foxl1(+) cells express stem cell markers such as Prom1 as well as differentiation markers such as Ck19 and Hnf4a. Thus, the Foxl1-Cre; RosaYFP model allows for easy isolation of adult hepatic progenitor cells that can be expanded and differentiated in culture. PMID- 21632826 TI - Prospective isolation of a bipotential clonogenic liver progenitor cell in adult mice. AB - The molecular identification of adult hepatic stem/progenitor cells has been hampered by the lack of truly specific markers. To isolate putative adult liver progenitor cells, we used cell surface-marking antibodies, including MIC1-1C3, to isolate subpopulations of liver cells from normal adult mice or those undergoing an oval cell response and tested their capacity to form bilineage colonies in vitro. Robust clonogenic activity was found to be restricted to a subset of biliary duct cells antigenically defined as CD45(-)/CD11b(-)/CD31(-)/MIC1 1C3(+)/CD133(+)/CD26(-), at a frequency of one of 34 or one of 25 in normal or oval cell injury livers, respectively. Gene expression analyses revealed that Sox9 was expressed exclusively in this subpopulation of normal liver cells and was highly enriched relative to other cell fractions in injured livers. In vivo lineage tracing using Sox9creER(T2)-R26R(YFP) mice revealed that the cells that proliferate during progenitor-driven liver regeneration are progeny of Sox9 expressing precursors. A comprehensive array-based comparison of gene expression in progenitor-enriched and progenitor-depleted cells from both normal and DDC (3,5-diethoxycarbonyl-1,4-dihydrocollidine or diethyl1,4-dihydro-2,4,6-trimethyl 3,5-pyridinedicarboxylate)-treated livers revealed new potential regulators of liver progenitors. PMID- 21632828 TI - Mast cells: bridging the gap between pre- and post-capillary pulmonary hypertension? PMID- 21632829 TI - Potential causes of increased long-term mortality after pneumonia. PMID- 21632830 TI - Macrolide-resistant Mycoplasma pneumoniae in paediatric pneumonia. PMID- 21632831 TI - Residential proximity to main roads and lung function in healthy young children. PMID- 21632832 TI - Severe hypersensitivity reaction as acute eosinophilic pneumonia and skin eruption induced by proguanil. PMID- 21632833 TI - A specific point-of-care screen for infectious pleural effusions using reagent strips. PMID- 21632834 TI - Response of automatic continuous positive airway pressure devices in a normal subject. PMID- 21632835 TI - The Asthma Control Questionnaire for children: still more questions than answers. PMID- 21632836 TI - Uneven use of noninvasive ventilation in acute respiratory failure in Europe. PMID- 21632837 TI - Sleep apnoea in Asians and Caucasians: comparing apples and oranges. PMID- 21632838 TI - The rationale for using hydroxycarbamide in the treatment of sickle cell disease" (Haematologica 2011;96:488-491). Reply. PMID- 21632839 TI - Increased incidence of central nervous system hemorrhages in patients with secondary acute promyelocytic leukemia after treatment of multiple sclerosis with mitoxantrone? PMID- 21632840 TI - Ring sideroblasts and sideroblastic anemias. PMID- 21632841 TI - Costs and consequences of immunosuppressive therapy in children with aplastic anemia. PMID- 21632842 TI - Treatment of acute myeloid leukemia in the elderly. PMID- 21632843 TI - The molecular basis of von Willebrand disease: the under investigated, the unexpected and the overlooked. PMID- 21632844 TI - Captopril treatment induces hyperplasia but inhibits myonuclear accretion following severe myotrauma in murine skeletal muscle. AB - The role of ANG II in skeletal muscle and satellite cell regulation is largely unknown. Cardiotoxin (CTX) was used to investigate whether muscle injury activates a local ANG II signaling system. Following injury, immunohistochelmistry (IHC) analysis revealed a robust increase in the intensity of angiotensinogen and angiotensin type 1 (AT(1)) receptor expression. As regeneration proceeded, however, AT(1) and angiotensinogen were downregulated. Nuclear accretion and fiber formation were also assessed during muscle regeneration in mice treated with captopril (an angiotensin-converting enzyme inhibitor). When ANG II formation was blocked through the use of captopril, we observed a significantly reduced accretion of nuclei into myofibers (-25%), while tibialis anterior total fiber number was significantly increased +37%. This phenotype appeared to be due to alterations in satellite cell differentiation kinetics; captopril treatment led to sustained mRNA expression of markers associated with quiescence and proliferation (Myf5, Pax7) and simultaneously delayed or inhibited the expression of myogenin. IHC staining supported these findings, revealing that captopril treatment resulted in a strong trend (P = 0.06) for a decrease in the proportion of myogenin-positive myoblasts. Furthermore, these observations were associated with a delay in muscle fiber maturation; captopril treatment resulted in sustained expression of embryonic myosin heavy chain. Collectively, these findings demonstrate that localized skeletal muscle angiotensin signaling is important to muscle fiber formation, myonuclear accretion, and satellite cell function. PMID- 21632845 TI - Methodological validation of the dynamic heterogeneity of muscle deoxygenation within the quadriceps during cycle exercise. AB - The conventional continuous wave near-infrared spectroscopy (CW-NIRS) has enabled identification of regional differences in muscle deoxygenation following onset of exercise. However, assumptions of constant optical factors (e.g., path length) used to convert the relative changes in CW-NIRS signal intensity to values of relative concentration, bring the validity of such measurements into question. Furthermore, to justify comparisons among sites and subjects, it is essential to correct the amplitude of deoxygenated hemoglobin plus myoglobin [deoxy(Hb+Mb)] for the adipose tissue thickness (ATT). We used two time-resolved NIRS systems to measure the distribution of the optical factors directly, thereby enabling the determination of the absolute concentrations of deoxy(Hb+Mb) simultaneously at the distal and proximal sites within the vastus lateralis (VL) and the rectus femoris muscles. Eight subjects performed cycle exercise transitions from unloaded to heavy work rates (>gas exchange threshold). Following exercise onset, the ATT-corrected amplitudes (A(p)), time delay (TD(p)), and time constant (tau(p)) of the primary component kinetics in muscle deoxy(Hb + Mb) were spatially heterogeneous (intersite coefficient of variation range for the subjects: 10-50 for A(p), 16-58 for TD(p), 14-108% for tau(p)). The absolute and relative amplitudes of the deoxy(Hb+Mb) responses were highly dependent on ATT, both within subjects and between measurement sites. The present results suggest that regional heterogeneity in the magnitude and temporal profile of muscle deoxygenation is a consequence of differential matching of O(2) delivery and O(2) utilization, not an artifact caused by changes in optical properties of the tissue during exercise or variability in the overlying adipose tissue. PMID- 21632846 TI - Endurance exercise training blunts the deleterious effect of high-fat feeding on whole body efficiency. AB - We recently showed that a week-long, high-fat diet reduced whole body exercise efficiency in sedentary men by >10% (Edwards LM, Murray AJ, Holloway CJ, Carter EE, Kemp GJ, Codreanu I, Brooker H, Tyler DJ, Robbins PA, Clarke K. FASEB J 25: 1088-1096, 2011). To test if a similar dietary regime would blunt whole body efficiency in endurance-trained men and, as a consequence, hinder aerobic exercise performance, 16 endurance-trained men were given a short-term, high-fat (70% kcal from fat) and a moderate carbohydrate (50% kcal from carbohydrate) diet, in random order. Efficiency was assessed during a standardized exercise task on a cycle ergometer, with aerobic performance assessed during a 1-h time trial and mitochondrial function later measured using (31)P-magnetic resonance spectroscopy. The subjects then underwent a 2-wk wash-out period, before the study was repeated with the diets crossed over. Muscle biopsies, for mitochondrial protein analysis, were taken at the start of the study and on the 5th day of each diet. Plasma fatty acids were 60% higher on the high-fat diet compared with moderate carbohydrate diet (P < 0.05). However, there was no change in whole body efficiency and no change in mitochondrial function. Endurance exercise performance was significantly reduced (P < 0.01), most probably due to glycogen depletion. Neither diet led to changes in citrate synthase, ATP synthase, or mitochondrial uncoupling protein 3. We conclude that prior exercise training blunts the deleterious effect of short-term, high-fat feeding on whole body efficiency. PMID- 21632847 TI - Do season and distribution affect thermal energetics of a hibernating bat endemic to the tropics and subtropics? AB - Although many tropical and subtropical areas experience pronounced seasonal changes in weather and food availability, few studies have examined and none have compared the thermal physiology and energetics of a hibernating mammal that is restricted to these regions. We quantified thermal energetics of northern long eared bats (Nyctophilus bifax; body mass ~10 g) during summer, winter, and spring from a subtropical habitat, and also during winter from a tropical habitat, to determine how N. bifax cope with climate and seasonal changes in weather. We captured bats in the wild and measured metabolic rates via open-flow respirometry. The basal metabolic rate of subtropical bats at an ambient temperature (T(a)) of 32.6 +/- 0.7 degrees C was 1.28 +/- 0.06 ml O(2).g(-1).h( 1) during both summer and winter, similar to other species of Nyctophilus. Resting metabolic rates below the thermoneutral zone increased similarly with decreasing T(a) during all seasons and in both regions. All individuals showed a high proclivity to enter torpor at T(a) values below the thermoneutral zone. Metabolic rates in torpid thermoconforming bats fell with T(a) and body temperature, and mean minimum metabolic rates during torpor were similar during all seasons and in both regions and as predicted from body mass in temperate zone hibernators. At very low T(a), torpid N. bifax thermoregulated, and this threshold T(a) differed significantly between subtropical (T(a) = 3.5 +/- 0.3 degrees C) and tropical (T(a) = 6.7 +/- 0.7 degrees C) individuals, but not between seasons. Our data show that thermal energetics of N. bifax do not vary seasonally and in many aspects are similar in tropical and subtropical bats; however, torpid individuals from the subtropics allow body temperature to fall to significantly lower values than those from the tropics. PMID- 21632848 TI - Estradiol potentiates hypothalamic vasopressin and oxytocin neuron activation and hormonal secretion induced by hypovolemic shock. AB - Estrogen receptors are located in important brain areas that integrate cardiovascular and hydroelectrolytic responses, including the subfornical organ (SFO) and supraoptic (SON) and paraventricular (PVN) nuclei. The aim of this study was to evaluate the influence of estradiol on cardiovascular and neuroendocrine changes induced by hemorrhagic shock in ovariectomized rats. Female Wistar rats (220-280 g) were ovariectomized and treated for 7 days with vehicle or estradiol cypionate (EC, 10 or 40 MUg/kg, sc). On the 8th day, animals were subjected to hemorrhage (1.5 ml/100 g for 1 min). Hemorrhage induced acute hypotension and bradycardia in the ovariectomized-oil group, but EC treatment inhibited these responses. We observed increases in plasma angiotensin II concentrations and decreases in plasma atrial natriuretic peptide levels after hemorrhage; EC treatment produced no effects on these responses. There were also increases in plasma vasopressin (AVP), oxytocin (OT), and prolactin levels after the induction of hemorrhage in all groups, and these responses were potentiated by EC administration. SFO neurons and parvocellular and magnocellular AVP and OT neurons in the PVN and SON were activated by hemorrhagic shock. EC treatment enhanced the activation of SFO neurons and AVP and OT magnocellular neurons in the PVN and SON and AVP neurons in the medial parvocellular region of the PVN. These results suggest that estradiol modulates the cardiovascular responses induced by hemorrhage, and this effect is likely mediated by an enhancement of AVP and OT neuron activity in the SON and PVN. PMID- 21632849 TI - Unique properties of muscularis mucosae smooth muscle in guinea pig urinary bladder. AB - The muscularis mucosae, a type of smooth muscle located between the urothelium and the urinary bladder detrusor, has been described, although its properties and role in bladder function have not been characterized. Here, using mucosal tissue strips isolated from guinea pig urinary bladders, we identified spontaneous phasic contractions (SPCs) that appear to originate in the muscularis mucosae. This smooth muscle layer exhibited Ca(2+) waves and flashes, but localized Ca(2+) events (Ca(2+) sparks, purinergic receptor-mediated transients) were not detected. Ca(2+) flashes, often in bursts, occurred with a frequency (~5.7/min) similar to that of SPCs (~4/min), suggesting that SPCs are triggered by bursts of Ca(2+) flashes. The force generated by a single mucosal SPC represented the maximal force of the strip, whereas a single detrusor SPC was ~3% of maximal force of the detrusor strip. Electrical field stimulation (0.5-50 Hz) evoked force transients in isolated detrusor and mucosal strips. Inhibition of cholinergic receptors significantly decreased force in detrusor and mucosal strips (at higher frequencies). Concurrent inhibition of purinergic and cholinergic receptors nearly abolished evoked responses in detrusor and mucosae. Mucosal SPCs were unaffected by blocking small-conductance Ca(2+)-activated K(+) (SK) channels with apamin and were unchanged by blocking large-conductance Ca(2+) activated K(+) (BK) channels with iberiotoxin (IbTX), indicating that SK and BK channels play a much smaller role in regulating muscularis mucosae SPCs than they do in regulating detrusor SPCs. Consistent with this, BK channel current density in myocytes from muscularis mucosae was ~20% of that in detrusor myocytes. These findings indicate that the muscularis mucosae in guinea pig represents a second smooth muscle compartment that is physiologically and pharmacologically distinct from the detrusor and may contribute to the overall contractile properties of the urinary bladder. PMID- 21632850 TI - Is visceral sympathoexcitation to heat stress dependent on activation of ionotropic excitatory amino acid receptors in the rostral ventrolateral medulla? AB - Acute heat stress activates visceral sympathetic nerve discharge (SND) in young rats, and the functional integrity of the rostral ventrolateral medulla (RVLM) is required for sustaining visceral sympathoexcitation during peak increases in internal body temperature (T(c)). However, RVLM mechanisms mediating SND activation to hyperthermia remain unknown. In the present study, we investigated the role of RVLM ionotropic excitatory amino acid receptors in mediating visceral SND activation to heat stress in anesthetized, young rats. The effects of bilateral RVLM kynurenic acid (Kyn; 2.7 and 5.4 nmol), saline, or muscimol (400 800 pmol) microinjections on renal SND and splenic SND responses to heat stress were determined at peak hyperthermia (T(c) 41.5 degrees C), during progressive hyperthermia (T(c) 40 degrees C), and at the initiation of heating (T(c) increased from 38 to 38.5 degrees C). RVLM Kyn microinjections did not reduce renal and splenic SND recorded during progressive or peak hyperthermia and did not attenuate SND activation at the initiation of heating. In fact, renal and splenic SND tended to be or were significantly increased following RVLM Kyn microinjections at the initiation of heating and during hyperthermia (40 and 41.5 degrees C). RVLM muscimol microinjections at 39, 40, and 41.5 degrees C resulted in immediate reductions in SND. These data indicate that RVLM ionotropic glutamate receptors are required for mediating visceral sympathoexcitation to acute heating and suggest that acute heating activates an RVLM ionotropic excitatory amino acid receptor dependent inhibitory input, which reduces the level of visceral SND to heating. PMID- 21632851 TI - Phase II study of abiraterone acetate in chemotherapy-naive metastatic castration resistant prostate cancer displaying bone flare discordant with serologic response. AB - PURPOSE: Abiraterone is an oral inhibitor of CYP17, which is essential for androgen biosynthesis. This multicenter study assessed its efficacy in patients with castration-resistant prostate cancer (CRPC), without prior chemotherapy or CYP17-targeted therapy, and frequency of bone scans discordant with prostate specific antigen (PSA) and clinical response. EXPERIMENTAL DESIGN: Thirty-three patients received abiraterone acetate 1,000 mg daily with prednisone 5 mg twice daily in continuous 28-day cycles. Patients were evaluated monthly for efficacy and safety. Bone scan flare was defined as the combination, after 3 months of therapy, of an interpreting radiologist's report indicating "disease progression" in context of a 50% or more decline in PSA level, with scan improvement or stability 3 months later. RESULTS: A 50% or more decline in PSA level at week 12 was confirmed in 22 of 33 (67%) patients. Declines in PSA level of 50% or more were seen in 26 of 33 (79%) patients. Undetectable PSA levels (<=0.1 ng/mL) occurred in 2 patients. Median time on therapy and time to PSA progression were 63 weeks and 16.3 months, respectively. Twenty-three patients were evaluable for bone scan flare. Progression was indicated in radiologist's report in 12 of 23 (52%), and 11 of 12 subsequently showed improvement or stability. As prospectively defined, bone scan flare was observed in 11 of 23 (48%) evaluable patients or 11 of 33 (33%) enrolled patients. Adverse events were typically grade 1/2 and consistent with prior published abiraterone reports. CONCLUSION: Clinical responses to abiraterone plus prednisone were frequent and durable in men with metastatic CRPC. Further investigation is needed to clarify the confounding effect of bone scan flare on patient management and interpretation of results. Clin Cancer Res; 17(14); 4854-61. (c)2011 AACR. PMID- 21632853 TI - Clinical significance of miR-146a in gastric cancer cases. AB - PURPOSE: The profiles of microRNAs change significantly in gastric cancer. MiR 146a is reported to be a tumor suppressor in pancreatic cancer, breast cancer, and prostate cancer. We investigated the clinical significance of miR-146a in gastric cancer, in particular focusing on hypothetical miR-146a target genes, such as epidermal growth factor receptor (EGFR) and interleukin-1 receptor associated kinase (IRAK1). EXPERIMENTAL DESIGN: We examined miR-146a levels in 90 gastric cancer samples by q-real-time (qRT)-PCR and analyzed the association between miR-146a levels and clinicopathologic factors and prognosis. The regulation of EGFR and IRAK1 by miR-146a was examined with miR-146a-transfected gastric cancer cells. Moreover, we analyzed the association between miR-146a levels and the G/C single nucleotide polymorphism (SNP) within pre-miR-146a seed sequences in 76 gastric cancer samples, using direct sequencing of genomic DNA. RESULTS: In 90 clinical samples of gastric cancer, miR-146a levels in cancer tissues were significantly lower than those in the corresponding noncancerous tissue (P < 0.001). Lower levels of miR-146a were associated with lymph node metastasis and venous invasion (P < 0.05). Moreover, a lower level of miR-146a was an independent prognostic factor for overall survival (P = 0.003). Ectopic expression of miR-146a inhibited migration and invasion and downregulated EGFR and IRAK1 expression in gastric cancer cells. In addition, G/C SNP within the pre miR-146a seed sequence significantly reduced miR-146a levels in the GG genotype compared with the CC genotype. CONCLUSIONS: MiR-146a contains an SNP, which is associated with mature miR-146a expression. MiR-146a targeting of EGFR and IRAK1 is an independent prognostic factor in gastric cancer cases. PMID- 21632852 TI - Myeloid biomarkers associated with glioblastoma response to anti-VEGF therapy with aflibercept. AB - PURPOSE: VEGF and infiltrating myeloid cells are known regulators of tumor angiogenesis and vascular permeability in glioblastoma. We investigated potential blood-based markers associated with radiographic changes to aflibercept, which binds VEGF and placental growth factor (PlGF) in patients with recurrent glioblastoma. EXPERIMENTAL DESIGN: In this single-arm phase II trial, aflibercept was given intravenously every two weeks until disease progression. Plasma and peripheral blood mononuclear cells were collected at baseline and 24 hours, 14 days, and 28 days posttreatment. Plasma cytokines and angiogenic factors were quantified by using ELISA and multiplex bead assays, and myeloid cells were assessed by flow cytometry in a subset of patients. RESULTS: Circulating levels of VEGF significantly decreased 24 hours after treatment with aflibercept, coincident with radiographic response observed by MRI. PlGF initially decreased 24 hours posttreatment but increased significantly by days 14 and 28. Lower baseline levels of PlGF, elevated baseline levels of CTACK/CCL27, MCP3/CCL7, MIF, and IP-10/CXCL10, and a decrease in VEGFR1(+) monocytes from baseline to 24 hours were all associated with improved response. Tumor progression was associated with increases in circulating matrix metalloproteinase 9. CONCLUSIONS: These data suggest that decreases in VEGF posttreatment are associated with radiographic response to aflibercept. Elevated baseline chemokines of monocyte lineage in responding patients supports a role for myeloid cells and chemokines as potential biomarkers and regulators of glioma angiogenesis. PMID- 21632854 TI - Molecular target characterization and antimyeloma activity of the novel, insulin like growth factor 1 receptor inhibitor, GTx-134. AB - PURPOSE: Therapeutic strategies that target insulin-like growth factor 1 receptor (IGF-1R) hold promise in a wide variety of cancers including multiple myeloma (MM). In this study, we describe GTx-134, a novel small-molecule inhibitor of IGF 1R and insulin receptor (IR) and characterized its antitumor activity in preclinical models of MM. EXPERIMENTAL DESIGN: The activity of GTx-134 as a single agent and in combination was tested in MM cell lines and primary patient samples. Downstream effector proteins and correlation with apoptosis was evaluated. Cytotoxcity in bone marrow stroma coculture experiments was assessed. Finally, the in vivo efficacy was evaluated in a human myeloma xenograft model. RESULTS: GTx-134 inhibited the growth of 10 of 14 myeloma cell lines (<5 MUmol/L) and induced apoptosis. Sensitivity to GTx-134 correlated with IGF-1R signal inhibition. Expression of MDR-1 and CD45 were associated with resistance to GTx 134. Coculture with insulin-growth factor-1 (IGF-1) or adherence to bone marrow stroma conferred modest resistance, but did not overcome GTx-134-induced cytotoxicity. GTx-134 showed in vitro synergies when combined with dexamethasone or lenalidomide. Further, GTx-134 enhanced the activity of PD173074, a fibroblast growth factor receptor 3 (FGFR3) inhibitor, against t(4;14) myeloma cells. Therapeutic efficacy of GTx-134 was shown against primary cells and xenograft tumors. Although dysregulation of glucose homeostasis was observed in GTx-134 treated mice, impairment of glucose tolerance was modest. CONCLUSIONS: These studies support the potential therapeutic efficacy of GTx-134 in MM. Further, they provide a rationale for clinical application in combination with established antimyeloma treatments and novel targeted therapies. PMID- 21632855 TI - Impact of clinical and pathologic features on tumor-infiltrating lymphocyte expansion from surgically excised melanoma metastases for adoptive T-cell therapy. AB - PURPOSE: Clinical trials on adoptive T-cell therapy (ACT) using expanded tumor infiltrating lymphocytes (TIL) have shown response rates of over 50% in refractory melanoma. However, little is known how clinical and pathologic features impact TIL outgrowth isolated from metastatic melanoma tumors. EXPERIMENTAL DESIGN: We analyzed the impact of clinical and pathologic features on initial TIL outgrowth in 226 consecutive patients undergoing tumor resection. Successful initial TIL outgrowth was defined as >=40 million viable lymphocytes harvested from all tumor fragments in a 5-week culture. To normalize for the different size of resected tumors and thus available tumor fragments, we divided the number of expanded TIL by the starting number of tumor fragments (TIL/fragment). RESULTS: Overall, initial TIL outgrowth was successful in 62% of patients, with patients <=30 years of age (94%; P = 0.01) and female patients (71% vs. 57% for males; P = 0.04) having the highest rate of success. Systemic therapy 30 days before tumor harvest negatively impacted initial TIL outgrowth compared to patients who never received systemic therapy (47% vs. 71%, P = 0.02). Biochemotherapy within 0 to 60 days of tumor harvest negatively impacted the initial TIL outgrowth with a success rate of only 16% (P < 0.0001). CONCLUSION: Parameters such as age, sex, and the type and timing of prior systemic therapy significantly affect the success rate of the initial TIL outgrowth from tumor fragments for ACT; these parameters may be helpful in selecting patients for melanoma ACT. PMID- 21632856 TI - Killing of chronic lymphocytic leukemia by the combination of fludarabine and oxaliplatin is dependent on the activity of XPF endonuclease. AB - PURPOSE: Chronic lymphocytic leukemia (CLL) resistant to fludarabine-containing treatments responds to oxaliplatin-based therapy that contains fludarabine. We postulated that a mechanism for this activity is the incorporation of fludarabine into DNA during nucleotide excision repair (NER) stimulated by oxaliplatin adducts. EXPERIMENTAL DESIGN: We analyzed CLL cell viability, DNA damage, and signaling pathways in response to treatment by fludarabine, oxaliplatin, or the combination. The dependency of the combination on oxaliplatin-induced DNA repair was investigated using siRNA in CLL cells or cell line models of NER deficiency. RESULTS: Synergistic apoptotic killing was observed in CLL cells after exposure to the combination in vitro. Oxaliplatin induced DNA synthesis in CLL cells, which was inhibited by fludarabine and was eliminated by knockdown of XPF, the NER 5'-endonuclease. Wild-type Chinese hamster ovarian cells showed synergistic killing after combination treatment, whereas only additive killing was observed in cells lacking XPF. Inhibition of repair by fludarabine in CLL cells was accompanied by DNA single-strand break formation. CLL cells initiated both intrinsic and extrinsic apoptotic pathways as evidenced by the loss of mitochondrial outer membrane potential and partial inhibition of cell death upon incubation with FasL antibody. CONCLUSIONS: The synergistic cell killing is caused by a mechanistic interaction that requires the initiation of XPF-dependent excision repair in response to oxaliplatin adducts, and the inhibition of that process by fludarabine incorporation into the repair patch. This combination strategy may be useful against other malignancies. PMID- 21632857 TI - Phase I study utilizing a novel antigen-presenting cell-targeted vaccine with Toll-like receptor stimulation to induce immunity to self-antigens in cancer patients. AB - PURPOSE: The use of tumor-derived proteins as cancer vaccines is complicated by tolerance to these self-antigens. Tolerance may be broken by immunization with activated, autologous, ex vivo generated and antigen-loaded, antigen-presenting cells (APC); however, targeting tumor antigen directly to APC in vivo would be a less complicated strategy. We wished to test whether targeted delivery of an otherwise poorly immunogenic, soluble antigen to APC through their mannose receptors (MR) would induce clinically relevant immunity. EXPERIMENTAL DESIGN: Two phase I studies were conducted with CDX-1307, a vaccine composed of human chorionic gonadotropin beta-chain (hCG-beta) fused to an MR-specific monoclonal antibody, administered either locally (intradermally) or systemically (intravenously) in patients with advanced epithelial malignancies. An initial dose escalation of single-agent CDX-1307 was followed by additional cohorts of CDX-1307 combined with granulocyte-macrophage colony-stimulating factor (GM-CSF) and the Toll-like receptor (TLR) 3 agonist polyinosinic-polycytidylic acid (poly ICLC) and TLR7/8 agonist resiquimod to activate the APC. RESULTS: CDX-1307 induced consistent humoral and T-cell responses to hCG-beta when coadministered with TLR agonists. Greater immune responses and clinical benefit, including the longest duration of stable disease, were observed with immunization combined with local TLR agonists. Immune responses were induced equally efficiently in patients with elevated and nonelevated levels of serum hCG-beta. Antibodies within the serum of vaccinated participants had tumor suppressive function in vitro. Toxicity consisted chiefly of mild injection site reactions. CONCLUSIONS: APC targeting and activation induce adaptive immunity against poorly immunogenic self antigens which has implications for enhancing the efficacy of cancer immunotherapy. PMID- 21632858 TI - Prognostic and predictive role of lactate dehydrogenase 5 expression in colorectal cancer patients treated with PTK787/ZK 222584 (vatalanib) antiangiogenic therapy. AB - PURPOSE: The Colorectal Oral Novel therapy For the Inhibition of angiogenesis and Retarding of Metastases (CONFIRM)-randomized trials, investigating the role of the VEGF-receptor inhibitor PTK787/ZK 222584 (vatalanib) in colorectal cancer (FOLFOX 4 +/- vatalanib), showed some benefit in patients with high serum lactate dehydrogenase (LDH) levels. Here, we investigated the expression of LDH5 (encoded entirely by the LDHA gene, regulated by the hypoxia inducible factors) in cancer tissues from patients recruited in the CONFIRM trials and relationship to response. EXPERIMENTAL DESIGN: Paraffin-embedded materials from 179 patients recruited in the CONFIRM trials were analyzed by immunohistochemistry for the expression of the LDH5 protein. Correlations with serum LDH, response, and survival were assessed. RESULTS: A significant association of tumor burden and of poor performance status (PS) with serum LDH was noted. Poor PS and high tumor LDH5 expression predicted for poor response rates. High tissue LDH5 was related to poor progression-free survival (PFS) only in the placebo group of patients, whereas the addition of vatalanib seemed to improved response and PFS in this subgroup. High serum LDH levels were linked with significantly poorer overall survival, which however was not sustained in multivariate analysis. CONCLUSIONS: Serum LDH and tissue LDH5 levels are complementary features that help to characterize the activity of LDH in colorectal cancer and have a potent value in predicting response to chemotherapy. The addition of vatalanib diminished the impact of LDH expression on the prognosis of patients. PMID- 21632859 TI - Is cytomegalovirus a therapeutic target in glioblastoma? AB - Several investigators have now demonstrated the expression of genes unique to cytomegalovirus (CMV) in malignant gliomas. Many of these genes promote oncogenesis, alter tumor microenvironment, and serve as immunologic targets. Is the level of CMV infection within tumor cells sufficient to drive important oncogenic or immunosuppressive processes? Can CMV serve as a target for therapeutic intervention? PMID- 21632860 TI - Increased detection sensitivity for KRAS mutations enhances the prediction of anti-EGFR monoclonal antibody resistance in metastatic colorectal cancer. AB - PURPOSE: KRAS mutations represent the main cause of resistance to anti-epidermal growth factor receptor (EGFR) monoclonal antibodies (MoAbs) in metastatic colorectal cancer (mCRC). We evaluated whether highly sensitive methods for KRAS investigation improve the accuracy of predictions of anti-EGFR MoAbs efficacy. EXPERIMENTAL DESIGN: We retrospectively evaluated objective tumor responses in mCRC patients treated with cetuximab or panitumumab. KRAS codons 12 and 13 were examined by direct sequencing, MALDI-TOF MS, mutant-enriched PCR, and engineered mutant-enriched PCR, which have a sensitivity of 20%, 10%, 0.1%, and 0.1%, respectively. In addition, we analyzed KRAS codon 61, BRAF, and PIK3CA by direct sequencing and PTEN expression by immunohistochemistry. RESULTS: In total, 111 patients were considered. Direct sequencing revealed mutations in codons 12 and 13 of KRAS in 43/111 patients (39%) and BRAF mutations in 9/111 (8%), with almost all of these occurring in nonresponder patients. Using highly sensitive methods, we identified up to 13 additional KRAS mutations compared with direct sequencing, all occurring in nonresponders. By analyzing PIK3CA and PTEN, we found that of these 13 patients, 7 did not show any additional alteration in the PI3K pathway. CONCLUSIONS: The application of highly sensitive methods for the detection of KRAS mutations significantly improves the identification of mCRC patients resistant to anti-EGFR MoAbs. PMID- 21632861 TI - High ALK receptor tyrosine kinase expression supersedes ALK mutation as a determining factor of an unfavorable phenotype in primary neuroblastoma. AB - PURPOSE: Genomic alterations of the anaplastic lymphoma kinase (ALK) gene have been postulated to contribute to neuroblastoma pathogenesis. This study aimed to determine the interrelation of ALK mutations, ALK expression levels, and clinical phenotype in primary neuroblastoma. EXPERIMENTAL DESIGN: The genomic ALK status and global gene expression patterns were examined in 263 primary neuroblastomas. Allele-specific ALK expression was determined by cDNA cloning and sequencing. Associations of genomic ALK alterations and ALK expression levels with clinical phenotypes and transcriptomic profiles were compared. RESULTS: Nonsynonymous point mutations of ALK were detected in 21 of 263 neuroblastomas (8%). Tumors with ALK mutations exhibited about 2-fold elevated median ALK mRNA levels in comparison with tumors with wild-type (WT) ALK. Unexpectedly, the WT allele was preferentially expressed in 12 of 21 mutated tumors. Whereas survival of patients with ALK mutated tumors was significantly worse as compared with the entire cohort of WT ALK patients, it was similarly poor in patients with WT ALK tumors in which ALK expression was as high as in ALK mutated neuroblastomas. Global gene expression patterns of tumors with ALK mutations or with high-level WT ALK expression were highly similar, and suggested that ALK may be involved in cellular proliferation in primary neuroblastoma. CONCLUSIONS: Primary neuroblastomas with mutated ALK exhibit high ALK expression levels and strongly resemble neuroblastomas with elevated WT ALK expression levels in both their clinical and molecular phenotypes. These data suggest that high levels of mutated and WT ALK mediate similar molecular functions that may contribute to a malignant phenotype in primary neuroblastoma. PMID- 21632862 TI - Engineering the brain tumor microenvironment enhances the efficacy of dendritic cell vaccination: implications for clinical trial design. AB - PURPOSE: Glioblastoma multiforme (GBM) is a deadly primary brain tumor. Clinical trials for GBM using dendritic cell (DC) vaccination resulted in antitumor immune responses. Herein, we tested the hypothesis that combining in situ (intratumoral) Ad-Flt3L/Ad-TK-mediated gene therapy with DC vaccination would increase therapeutic efficacy and antitumor immunity. EXPERIMENTAL DESIGN: We first assessed the immunogenicity of tumor lysates generated by Ad-TK (+GCV), temozolomide (TMZ), or freeze/thawing cycles (FTC) in a syngeneic brain tumor model. We also assessed phenotypic markers, cytokine release, and phagocytosis of bone marrow-derived DCs generated by fms-like tyrosine kinase 3 ligand (Flt3L) + IL-6 or by granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL) 4. Inhibition of tumor progression and production of anti-GBM antibodies was assessed following vaccination with (i) tumor cell lysates, (ii) DCs generated with either Flt3L/IL-6 or GM-CSF/IL-4 loaded with either Ad-TK/GCV , TMZ-, or FTC-generated tumor lysates, or (iii) DCs in combination with in situ Ad-Flt3L/Ad-TK gene therapy. RESULTS: DCs loaded with tumor cell lysates generated with either Ad-TK/GCV or TMZ led to increased levels of phagocytosis, therapeutic efficacy, and humoral immune response. In situ immunogene therapy in combination with DC vaccination led to brain tumor regression and long-term survival in about 90% of animals, a significant increase when compared with either therapy alone. CONCLUSIONS: Our results indicate that modifying the tumor microenvironment using intratumoral Ad-Flt3L/Ad-TK-mediated gene therapy potentiates therapeutic efficacy and antitumor immunity induced by DC vaccination. These data support novel phase I clinical trials to assess the safety and efficacy of this combined approach. PMID- 21632863 TI - Mediastinal Castleman's disease mimicking thoracic paravertebral schwannoma. AB - A 51-year-old female underwent resection of a solid lesion in the posterior mediastinum, preoperatively interpreted at imaging as thoracic schwannoma, requiring double sequential surgical procedure to be resected. The histologic examination of the resected mass diagnosed a hyaline-vascular Castleman's disease. PMID- 21632864 TI - Outcomes of total aortic arch replacement with coronary artery bypass grafting. AB - There are few reports regarding outcomes of total aortic arch replacement (TAAR) with concomitant coronary artery bypass grafting (CABG). This study aimed to analyze outcomes after total arch replacement with CABG in our institute. Between July 2006 and December 2010, 126 patients underwent elective total arch replacement with or without CABG. Of these patients, 56 had concomitant CABG. The mean age was 73.9+/-5.6 years. Fifteen patients were female. Six patients had chronic aortic dissection, and two patients had previous aortic operations. Total arch replacement was performed with selective cerebral perfusion and the elephant trunk procedure. The mean number of coronary anastomoses was 1.9+/-1.0. An internal thoracic artery (ITA) and saphenous vein graft were used in 37 (66.1%) and 47 (83.9%) patients, respectively. Only the anastomosis between the in-situ ITA and the left anterior descending artery (LADA) was performed after declamping the aorta using the on-pump beating technique. Operative mortality was 3.6%. Stroke occurred in 7.1% of patients. Early graft patency of bypass grafts was 100%. Composite outcomes of mortality and the rate of major complications of patients with CABG was 19.6%. TAAR with CABG can be safely performed with favorable outcomes. PMID- 21632865 TI - What size of left atrium significantly impairs the success of maze surgery for atrial fibrillation? AB - A best evidence topic in cardiothoracic surgery was written according to a structured protocol. The question addressed was in adults with atrial fibrillation (AF), what preoperative size of left atrium impairs maze surgery success in terms of recurrence of AF. Altogether 422 papers were found using the reported search, of which 12 represented the best evidence to answer the clinical question. The authors, journal. date and country of publication, patient group studied, study type, relevant outcomes and results of these papers are tabulated. Nine of 12 papers found that preoperative left atrial (LA) size was significantly larger in patients who experienced recurrent AF. When left atrial diameter (LAD) was found to be statistically different between sinus conversion and recurrent AF groups the mean LAD was consistently >60 mm in the recurrent AF group, whereas it was <60 mm in all but one of the sinus conversion groups. In terms of a cut-off value for LA size, a left atrial volume index >135 ml/m(2) was found to confer 100% specificity for maze failure and a LAD >60 mm was found to be 100% sensitive for maze failure. A preoperative LAD <48.3 mm was shown in one study to be 100% sensitive for sinus conversion by the maze procedure. Despite much evidence highlighting preoperative LAD as a risk factor for maze failure, relatively few studies seek to define a definitive cut-off value for LA size beyond which the risks of the procedure (such as bleeding, infection or stroke) outweigh the chance of sinus recovery. We conclude that since mean preoperative LAD in AF groups is consistently over 60 mm caution should be exercised when offering these patients the maze procedure. Furthermore, the relationship between preoperative LAD and maze failure appears continuous and so patients should be counselled as to their increased risk of failure the further they deviate from a LAD of 60 mm. There is some evidence for and no available evidence to the contrary that a LAD < 43 mm is associated with complete maze success. Hence, these patients should be offered the maze procedure unless there are alternate strong contraindications. PMID- 21632866 TI - Thoracoscopic removal of a suture needle from the posterior pericardium after coronary artery bypass grafting. AB - We demonstrate a minimally-invasive thoracoscopic approach [video-assisted thoracic surgery (VATS)] for removal of a retained pericardial suture needle after standard coronary artery bypass grafting (CABG) surgery. A 46-year-old male presented with unstable angina. His workup demonstrated significant coronary artery disease for which he underwent a six vessel CABG, including entering the left chest for preparation of the left internal mammary artery (LIMA). At seven weeks, a postoperative chest X-ray demonstrated a foreign body (suture needle) present in the cardiac silhouette. Further computed tomography (CT)-scan imaging confirmed the suture needle to be localized in the left inferior-posterior pericardium. The patient underwent a left VATS exploration for removal of the suture needle. The pericardial suture needle was successfully retrieved thoracoscopically. The chest tube was removed on the first postoperative day and the patient was discharged to home on the second postoperative day. The patient's postoperative course and recovery were uneventful. A minimally-invasive approach can be undertaken for the removal of a foreign body even after prior open chest surgery, avoiding the associated morbidity of a repeat sternotomy. PMID- 21632867 TI - Neo-coarctation after the arterial switch operation. AB - Neo-coarctation following arterial switch operation (ASO) for transposition of the great arteries (TGA) is a complication that is not regularly described, but may occur. We describe five patients who developed a neo-coarctation after operation. They were diagnosed with TGA, either with or without ventricular septal defect without signs or symptoms of a coarctation. Except for one patient, all patients were reoperated for a neo-coarctation within one year after the ASO. Several explanations are discussed as a possible cause for this phenomenon. PMID- 21632868 TI - Mechanisms underlying the renoprotective effect of GABA against ischemia/reperfusion-induced renal injury in rats. AB - The excitation of the renal sympathetic nervous system plays an important role in the development of ischemic acute kidney injury (AKI) in rats. We have reported that intravenous treatment with GABA has preventive effects on ischemia/reperfusion (I/R)-induced renal dysfunction with histological damage in rats. However, detailed mechanisms of the action of GABA on the renal injury were still unknown. Therefore, in the present study, we aimed to clarify the detailed mechanisms of GABA in ischemic AKI in rats. Ischemic AKI was induced by clamping the left renal artery and vein for 45 min. Thereafter, the kidney was reperfused to produce I/R-induced injury. Intravenous or intracerebroventricular treatment with 3-[[[(3,4-dichlorophenyl)methyl]amino]propyl] diethoxymethyl) phosphinic acid (CGP52432), a GABA(B) receptor antagonist, abolished the suppressive effects of intravenously applied GABA on enhanced renal sympathetic nerve activity during ischemia, leading to the elimination of the renoprotective effects of GABA. Intracerebroventricular treatment with GABA or intravenous treatment with baclofen, a selective GABA(B) receptor agonist, prevented I/R-induced renal injury equivalent to intravenous treatment with GABA. However, intravenous treatment with bicuculline, a GABA(A) receptor antagonist, failed to affect the preventive effects of GABA on ischemic AKI. Therefore, we demonstrated the novel finding that the preventive effect of GABA on ischemic AKI through the suppression of enhanced renal sympathetic nerve activity induced by renal ischemia is presumably mediated via GABA(B) receptor stimulation in the central nervous system rather than peripheral GABA(B) receptor. PMID- 21632869 TI - Characterization of a sphingosine 1-phosphate receptor antagonist prodrug. AB - Sphingosine 1-phosphate (S1P) is a phospholipid that binds to a set of G protein coupled receptors (S1P(1)-S1P(5)) to initiate an array of signaling cascades that affect cell survival, differentiation, proliferation, and migration. On a larger physiological scale, the effects of S1P on immune cell trafficking, vascular barrier integrity, angiogenesis, and heart rate have also been observed. An impetus for the characterization of S1P-initiated signaling effects came with the discovery that FTY720 [fingolimod; 2-amino-2-(2-[4-octylphenyl]ethyl)-1,3 propanediol] modulates the immune system by acting as an agonist at S1P(1). In the course of structure-activity relationship studies to better understand the functional chemical space around FTY720, we discovered conformationally constrained FTY720 analogs that behave as S1P receptor type-selective antagonists. Here, we present a pharmacological profile of a lead S1P(1/3) antagonist prodrug, 1-(hydroxymethyl)-3-(3-octylphenyl)cyclobutane (VPC03090). VPC03090 is phosphorylated by sphingosine kinase 2 to form the competitive antagonist species 3-(3-octylphenyl)-1-(phosphonooxymethyl)cyclobutane (VPC03090 P) as observed in guanosine 5'-O-(3-[(35)S]thio)triphosphate binding assays, with effects on downstream S1P receptor signaling confirmed by Western blot and calcium mobilization assays. Oral dosing of VPC03090 results in an approximate 1:1 phosphorylated/alcohol species ratio with a half-life of 30 h in mice. Because aberrant S1P signaling has been implicated in carcinogenesis, we applied VPC03090 in an immunocompetent mouse mammary cancer model to assess its antineoplastic potential. Treatment with VPC03090 significantly inhibited the growth of 4T1 primary tumors in mice. This result calls to attention the value of S1P receptor antagonists as not only research tools but also potential therapeutic agents. PMID- 21632870 TI - High-resolution structural insights on the sugar-recognition and fusion tag properties of a versatile beta-trefoil lectin domain from the mushroom Laetiporus sulphureus. AB - In this work, we analyzed at high resolution the sugar-binding mode of the recombinant N-terminal ricin-B domain of the hemolytic protein LSLa (LSL(150)) from the mushroom Laetiporus sulphureus and also provide functional in vitro evidence suggesting that, together with its putative receptor-binding role, this module may also increase the solubility of its membrane pore-forming partner. We first demonstrate that recombinant LSL(150) behaves as an autonomous folding unit and an active lectin. We have determined its crystal structure at 1.47 A resolution and also that of the [LSL(150):(lactose)beta, gamma)] binary complex at 1.67 A resolution. This complex reveals two lactose molecules bound to the beta and gamma sites of LSL(150), respectively. Isothermal titration calorimetry indicates that LSL(150) binds two lactoses in solution with highly different affinities. Also, we test the working hypothesis that LSL(150) exhibits in vivo properties typical of solubility tags. With this aim, we have fused an engineered version of LSL(150) (LSL(t)) to the N-terminal end of various recombinant proteins. All the designed LSL(150)-tagged fusion proteins were successfully produced at high yield, and furthermore, the target proteins were purified by a straightforward affinity procedure on agarose-based matrices due to the excellent properties of LSL(150) as an affinity tag. An optimized protocol for target protein purification was devised, which involved removal of the LSL(150) tag through in-column cleavage of the fusion proteins with His(6)-tagged TEV endoprotease. These results permitted to set up a novel, lectin-based system for production and purification of recombinant proteins in E. coli cells with attractive biotechnological applications. PMID- 21632871 TI - Mutational probing of the forkhead domain of the transcription factor FOXL2 provides insights into the pathogenicity of naturally occurring mutations. AB - Mutations of the transcription factor FOXL2, involved in cranio-facial and ovarian development, lead to the Blepharophimosis Syndrome. Here, we have systematically replaced the amino acids of the helices of the forkhead domain (FHD) of FOXL2 by glycine residues to assess the impact of such substitutions. A number of mutations lead to protein mislocalization, aggregation and to partial or complete loss of transactivation ability on a series of luciferase reporter systems. To rationalize the results of this glycine mutation scan, we have modeled the structure of the FHD by comparison with crystallographic data available for other FHDs. We failed to detect a clear-cut correlation between protein mislocalization or aggregation and the position of the mutation. However, we found that the localization of the side chain of each amino acid strongly correlated with the impact of its mutation on FOXL2 transactivation capacity. Indeed, when the side chains of the amino acids involved in the helices of the forkhead are supposed to point towards the hydrophobic core formed by the three main helices, a loss of function was observed. On the contrary, if the side chains point outward the hydrophobic core, protein function was preserved. The extension of this analysis to natural mutants shows that a similar correlation can be found for BPES mutations associated or not with ovarian dysfunction. Our findings reveal new insights into the molecular effects of FOXL2 mutations affecting the FHD, which represent two-thirds of intragenic mutations, and provide the first predictive tool of their effects. PMID- 21632872 TI - Psychogenic nonepileptic seizures (pseudoseizures). PMID- 21632873 TI - Normal pubertal development: Part I: The endocrine basis of puberty. PMID- 21632874 TI - Speech and language development: monitoring process and problems. PMID- 21632875 TI - Acid-base disorders. PMID- 21632877 TI - Swelling and redness of the fourth toe in a 3-month-old infant. PMID- 21632878 TI - Index of Suspicion. Case 1: Status epilepticus, hypertension, and tachycardia in a 5-year-old boy. Case 2: Cardiopulmonary arrest during gymnastics practice in a teenage girl. Case 3: Acute renal failure in a teenage boy who has autism and pica. PMID- 21632879 TI - Contemporary cardiogenesis: new insights into heart development. PMID- 21632880 TI - Induction of MesP1 by Brachyury(T) generates the common multipotent cardiovascular stem cell. AB - AIMS: Our recent work demonstrated that common cardiovascular progenitor cells are characterized and induced by the expression of the transcription factor mesoderm posterior1 (MesP1) in vertebrate embryos and murine embryonic stem cells. As the proliferative potential of stem cell-derived cardiomyocytes is limited, it is crucial to understand how MesP1 expression is mediated in order to achieve reasonable and reliable yields for novel stem cell-based therapeutic options. As potential upstream regulators of MesP1, we therefore analysed Eomes and Brachyury(T), which had been controversially discussed as being crucial for cardiovasculogenic lineage formation. METHODS AND RESULTS: Wild-type and transgenic murine embryonic stem cell lines, mRNA analyses, embryoid body formation, and cell sorting revealed that the MesP1 positive population emerges from the Brachyury(T) positive fraction. In situ hybridizations using wild-type mouse embryos confirmed that Brachyury(T) colocalises with MesP1 in vivo. Likewise, shRNA-based loss of Brachyury(T) causes a dramatic decrease in MesP1 expression accompanied by reduced cardiac markers in differentiating embryonic stem cells, which is reflected in vivo via in situ hybridizations using Brachyury(T) knock-out embryos where MesP1 mRNA is greatly abolished. We finally defined a 3.4 kb proximal MesP1-promoter fragment which is directly bound and activated by Brachyury(T) via a T responsive element as shown via bandshift, chromatin immuneprecipitation, and reporter assays. CONCLUSION: Our work contributes to the understanding of the earliest cardiovasculogenic events and may become an important prerequisite for cell therapy, tissue engineering, and pharmacological testing in the culture dish using pluripotent stem cell-derived as well as directly reprogrammed cardiovascular cell types. PMID- 21632882 TI - p2y5/LPA(6) attenuates LPA(1)-mediated VE-cadherin translocation and cell-cell dissociation through G(12/13) protein-Src-Rap1. AB - AIMS: We investigated the mechanisms of action of lysophosphatidic acid (LPA) to regulate vascular endothelial (VE)-cadherin dynamics and cell-cell contact. METHODS AND RESULTS: While a low concentration of LPA stimulated VE-cadherin internalization and subsequent cell-cell dissociation, a high concentration of LPA masked the disruptive actions on VE-cadherin and protected the barrier function in human vascular endothelial cells. Knockdown experiments of major LPA receptor subtypes, i.e. LPA(1) and p2y5 (also termed LPA(6)), with their specific small interfering RNAs, showed that LPA(1) and LPA(6) mediate the LPA-induced disruptive and protective actions on barrier integrity, respectively. LPA(6) mediated tube formation, reflecting stabilization of barrier integrity, was confirmed by in vitro angiogenesis assay. The LPA(1)-mediated disruptive actions were inhibited by pertussis toxin, dominant-negative Rac1, and inhibitors for c Jun NH(2)-terminal kinase (JNK) and p38 mitogen-activated protein kinase (p38MAPK), but not by dominant-negative RhoA. In contrast, the LPA(6)-mediated protective actions were associated with activation of Src and Rap1 and attenuated by abrogation of their activities. Further characterization showed that Rap1 is located downstream of Src and dependent on C3G, a Rap1 guanine nucleotide exchange factor. Finally, an LPA antagonist significantly inhibited lactic acid induced limb lesions in vivo, which may be attributed to dysfunction of endothelial cells. CONCLUSION: LPA induced disruption and protection of VE cadherin integrity through LPA(1)-G(i) protein-Rac1-JNK/p38MAPK and LPA(6) G(12/13) protein-Src-C3G-Rap1 pathways, respectively. PMID- 21632881 TI - Cardiac-specific mindin overexpression attenuates cardiac hypertrophy via blocking AKT/GSK3beta and TGF-beta1-Smad signalling. AB - AIMS: Mindin is a secreted extracellular matrix protein, an integrin ligand, and an angiogenesis inhibitor, other examples of which are all key players in the progression of cardiac hypertrophy. However, its function during cardiac hypertrophy remains unclear. This study was aimed to identify the effect of mindin on cardiac hypertrophy and the underlying mechanisms. METHODS AND RESULTS: A significant down-regulation of mindin expression was observed in human failing hearts. To further investigate the role of mindin in cardiac hypertrophy, we used cultured neonatal rat cardiomyocytes with gain and loss of mindin function and cardiac-specific Mindin-overexpressing transgenic (TG) mice. In cultured cardiomyocytes, mindin negatively regulated angiotensin II (Ang II)-mediated hypertrophic growth, as detected by [(3)H]-Leucine incorporation, cardiac myocyte area, and hypertrophic marker protein levels. Cardiac hypertrophy in vivo was produced by aortic banding (AB) or Ang II infusion in TG mice and their wild-type controls. The extent of cardiac hypertrophy was evaluated by echocardiography as well as by pathological and molecular analyses of heart samples. Mindin overexpression in the heart markedly attenuated cardiac hypertrophy, fibrosis, and left ventricular dysfunction in mice in response to AB or Ang II. Further analysis of the signalling events in vitro and in vivo indicated that these beneficial effects of mindin were associated with the interruption of AKT/glycogen synthase kinase 3beta (GSK3beta) and transforming growth factor (TGF)-beta1-Smad signalling. CONCLUSION: The present study demonstrates for the first time that mindin serves as a novel mediator that protects against cardiac hypertrophy and the transition to heart failure by blocking AKT/GSK3beta and TGF beta1-Smad signalling. PMID- 21632883 TI - Syndecan-4 signalling inhibits apoptosis and controls NFAT activity during myocardial damage and remodelling. AB - AIMS: Myocardial infarction (MI) results in acute impairment of left ventricular (LV) function through the initial development of cardiomyocyte death and subsequent progression of LV remodelling. The expression of syndecan-4 (Sdc4), a transmembrane proteoglycan, is up-regulated after MI, but its function in the heart remains unknown. Here, we characterize the effects of Sdc4 deficiency in murine myocardial ischaemia and permanent infarction. METHODS AND RESULTS: Targeted deletion of Sdc4 (Sdc4(-/-)) leads to increased myocardial damage after ischaemic-reperfusion injury due to enhanced cardiomyocyte apoptosis associated with reduced activation of extracellular signal-regulated kinase in cardiomyocytes in vitro and in vivo. After ischaemic-reperfusion injury and permanent infarction, we observed an increase in cardiomyocyte area, nuclear translocation of nuclear factor of activated T cells (NFAT), and transcription of the NFAT target rcan1.4 in wild-type mice. NFAT pathway activation was enhanced in Sdc4(-/-) mice. In line with the in vivo data, NFAT activation and hypertrophy occurs in isolated cardiomyocytes with reduced Sdc4 expression during phenylephrine stimulation in vitro. Despite the initially increased myocardial damage, echocardiography revealed improved LV geometry and function in Sdc4(-/-) mice 7 days after MI. CONCLUSION: Interception of the Sdc4 pathway enhances infarct expansion and hypertrophic remodelling during early infarct healing in ischaemic-reperfusion injury and permanent infarction mouse models and exerts net beneficial effects on LV function. PMID- 21632884 TI - Deletion of the metabolic transcriptional coactivator PGC1beta induces cardiac arrhythmia. AB - AIMS: Peroxisome proliferator-activated receptor-gamma coactivators PGC1alpha and PGC1beta modulate mitochondrial biogenesis and energy homeostasis. The function of these transcriptional coactivators is impaired in obesity, insulin resistance, and type 2 diabetes. We searched for transcriptomic, lipidomic, and electrophysiological alterations in PGC1beta(-/-) hearts potentially associated with increased arrhythmic risk in metabolic diseases. METHODS AND RESULTS: Microarray analysis in mouse PGC1beta(-/-) hearts confirmed down-regulation of genes related to oxidative phosphorylation and the electron transport chain and up-regulation of hypertrophy- and hypoxia-related genes. Lipidomic analysis showed increased levels of the pro-arrhythmic and pro-inflammatory lipid, lysophosphatidylcholine. PGC1beta(-/-) mouse electrocardiograms showed irregular heartbeats and an increased incidence of polymorphic ventricular tachycardia following isoprenaline infusion. Langendorff-perfused PGC1beta(-/-) hearts showed action potential alternans, early after-depolarizations, and ventricular tachycardia. PGC1beta(-/-) ventricular myocytes showed oscillatory resting potentials, action potentials with early and delayed after-depolarizations, and burst firing during sustained current injection. They showed abnormal diastolic Ca(2+) transients, whose amplitude and frequency were increased by isoprenaline, and Ca(2+) currents with negatively shifted inactivation characteristics, with increased window currents despite unaltered levels of CACNA1C RNA transcripts. Inwardly and outward rectifying K(+) currents were all increased. Quantitiative RT-PCR demonstrated increased SCN5A, KCNA5, RYR2, and Ca(2+)-calmodulin dependent protein kinase II expression. CONCLUSION: PGC1beta(-/-) hearts showed a lysophospholipid-induced cardiac lipotoxicity and impaired bioenergetics accompanied by an ion channel remodelling and altered Ca(2+) homeostasis, converging to produce a ventricular arrhythmic phenotype particularly during adrenergic stress. This could contribute to the increased cardiac mortality associated with both metabolic and cardiac disease attributable to lysophospholipid accumulation. PMID- 21632885 TI - MicroRNAs as regulators of signal transduction in urological tumors. AB - BACKGROUND: MicroRNAs (miRNAs) are short noncoding RNAs that have been shown to play pivotal roles in carcinogenesis. In the past decade, miRNAs have been the focus of much research in oncology, and there are great expectations for their utility as cancer biomarkers and therapeutic targets. CONTENT: In this review we examine how miRNAs can regulate signal transduction pathways in urological tumors. We performed in silico target prediction using TargetScan 5.1 to identify the signal transduction targets of miRNA, and we summarize the experimental evidence detailing miRNA regulation of pathways analyzed herein. SUMMARY: miRNAs, which have been shown to be dysregulated in bladder, prostate, and renal cell cancer, are predicted to target key proteins in signal transduction. Because androgen receptor signaling is a major regulator of prostate cancer growth, its regulation by miRNAs has been well described. In addition, members of the phosphatidylinositol 3-kinase/Akt (RAC-alpha serine/threonine-protein kinase) signaling pathway have been shown to be susceptible to miRNA regulation. In contrast, there are very few studies on the impact of miRNA regulation on signaling by VHL (von Hippel-Lindau tumor suppressor) and vascular endothelial growth factor in renal cell carcinoma or by fibroblast growth factor receptor 3 and p53 in bladder cancer. Many miRNAs are predicted to target important signaling pathways in urological tumors and are dysregulated in their respective cancer types; a systematic overview of miRNA regulation of signal transduction in urological tumors is pending. The identification of these regulatory networks might lead to novel targeted cancer therapies. In general, the targeting of miRNAs is a valuable approach to cancer therapy, as has been shown recently for various types of cancer. PMID- 21632887 TI - Comments on geographic maldistribution of primary care for children. PMID- 21632888 TI - Clinical significance of interleukin-2/gamma interferon ratios in Mycobacterium tuberculosis-specific T-cell signatures. AB - The simultaneous determination of interleukin-2 (IL-2) and gamma interferon (IFN gamma) in QuantiFERON-TB test plasma supernatants permitted the detection of shifts in Mycobacterium tuberculosis-specific T-cell signatures. A subset of the 84 subjects tested revealed a significantly elevated IL-2/IFN-gamma ratio, which may be a marker for the successful elimination of M. tuberculosis infection. PMID- 21632889 TI - Comparative Immunogenicities of full-length Plasmodium falciparum merozoite surface protein 3 and a 24-kilodalton N-terminal fragment. AB - Recombinant Plasmodium falciparum merozoite surface protein 3 (PfMSP3F) and a 24 kDa fragment from its N terminus (MSP3N) that includes the essential conserved domain, which elicits the maximum antibody (Ab)-dependent cellular inhibition (ADCI), were expressed as soluble proteins in Escherichia coli. Both proteins were found to be stable in both soluble and lyophilized forms. Immunization with MSP3F and MSP3N formulated separately with two human-compatible adjuvants, aluminum hydroxide (Alhydrogel) and Montanide ISA 720, produced significant antibody responses in mice and rabbits. Polyclonal Abs against both antigens recognized native MSP3 in the parasite lysate. These two Abs also recognized two synthetic peptides, previously characterized to possess B cell epitopes from the N-terminal region. Antibody depletion assay showed that most of the IgG response is directed toward the N-terminal region of the full protein. Anti-MSP3F and anti MSP3N rabbit antibodies did not inhibit merozoite invasion or intraerythrocytic development but significantly reduced parasitemia in the presence of human monocytes. The ADCI demonstrated by anti-MSP3N antibodies was comparable to that exhibited by anti-MSP3F antibodies (both generated in rabbit). These results suggest that the N-terminal fragment of MSP3 can be considered a vaccine candidate that can form part of a multigenic vaccine against malaria. PMID- 21632890 TI - Evaluation of oral immunization with recombinant avian influenza virus HA1 displayed on the Lactococcus lactis surface and combined with the mucosal adjuvant cholera toxin subunit B. AB - The development of safe and efficient avian influenza vaccines for human and animal uses is essential for preventing virulent outbreaks and pandemics worldwide. In this study, we constructed a recombinant (pgsA-HA1 gene fusion) Lactococcus lactis strain that expresses and displays the avian influenza virus HA1 antigens on its surface. The vectors were administered by oral delivery with or without the addition of cholera toxin subunit B (CTB). The resulting immune responses were analyzed, and the mice were eventually challenged with lethal doses of H5N1 viruses. Significant titers of hemagglutinin (HA)-specific serum IgG and fecal IgA were detected in the group that also received CTB. Cellular immunities were also shown in both cell proliferation and gamma interferon (IFN gamma) enzyme-linked immunospot (ELISpot) assays. Most importantly, the mice that received the L. lactis pgsA-HA1 strain combined with CTB were completely protected from lethal challenge of the H5N1 virus. These findings support the further development of L. lactis-based avian influenza virus vaccines for human and animal uses. PMID- 21632891 TI - KSAC, the first defined polyprotein vaccine candidate for visceral leishmaniasis. AB - A subunit vaccine using a defined antigen(s) may be one effective solution for controlling leishmaniasis. Because of genetic diversity in target populations, including both dogs and humans, a multiple-antigen vaccine will likely be essential. However, the cost of a vaccine to be used in developing countries must be considered. We describe herein a multiantigen vaccine candidate comprised of antigens known to be protective in animal models, including dogs, and to be recognized by humans immune to visceral leishmaniasis. The polyprotein (KSAC) formulated with monophosphoryl lipid A, a widely used adjuvant in human vaccines, was found to be immunogenic and capable of inducing protection against Leishmania infantum, responsible for human and canine visceral leishmaniasis, and against L. major, responsible for cutaneous leishmaniasis. The results demonstrate the feasibility of producing a practical, cost-effective leishmaniasis vaccine capable of protecting both humans and dogs against multiple Leishmania species. PMID- 21632892 TI - Evaluation of a new DNA test for detection of carcinogenic human papillomavirus. AB - Using archived specimens, we evaluated a new automated real-time PCR assay (BD Diagnostics) that detects all carcinogenic human papillomaviruses (HPV) and provides HPV genotyping for seven of them, including HPV16 and HPV18, the two most carcinogenic HPV genotypes. We found comparable results with Hybrid Capture 2 (HC2) for detection of carcinogenic HPV (n = 473) and with Linear Array and Line Blot Assay (n = 371) for detection of individual HPV genotypes. PMID- 21632893 TI - Adherence to CLSI recommendations for testing of Staphylococcus aureus isolates in Louisiana hospitals: report of a clinical failure and results of a questionnaire study. AB - We report a case of failure of clindamycin therapy due to inducible clindamycin resistance. We surveyed and found that only 52% of reporting hospitals in the state of Louisiana were performing the D test for inducible clindamycin resistance according to guidelines recommended by the Clinical and Laboratory Standards Institute (formerly the National Committee for Clinical Laboratory Standards). PMID- 21632894 TI - Utility of matrix-assisted laser desorption ionization-time of flight mass spectrometry following introduction for routine laboratory bacterial identification. AB - Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) was evaluated prospectively in a diagnostic laboratory. Nine hundred twenty-seven organisms were tested in triplicate; 2,351/2,781 (85%) species and 2,681/2,781 (96%) genus identifications were correct. Known issues such as the misidentification of alpha-hemolytic streptococci as Streptococcus pneumoniae were easily corrected. Identifications cost AUD$0.45 per isolate and were available in minutes. MALDI-TOF MS is rapid, accurate, and inexpensive. PMID- 21632895 TI - Practical approach for reliable detection of AmpC beta-lactamase-producing Enterobacteriaceae. AB - In this prospective study all Enterobacteriaceae isolates (n = 2,129) recovered in the clinical microbiology laboratory during October 2009 to April 2010 were analyzed for AmpC production. Clinical and Laboratory Standards Institute (CLSI) cefoxitin and cefotetan susceptibility breakpoints and CLSI critical ESBL diameters were used to screen for potential AmpC producers. In total, 305 isolates (211 potential AmpC producers and 94 AmpC screen-negative isolates as a control group) were further analyzed by multiplex PCR for the detection of plasmid-encoded ampC beta-lactamase genes and by ampC promoter sequence analysis (considered as the gold standard). Cefoxitin and cefotetan were assessed as primary screening markers. The sensitivities of cefoxitin and cefotetan for the detection of AmpC production were 97.4 and 52.6%, respectively, and the specificities were 78.7 and 99.3%, respectively. As a phenotypic confirmation test, the Etest AmpC and the cefoxitin-cloxacillin double-disk synergy method (CC DDS) were compared. The sensitivities for the Etest AmpC and the CC-DDS method were 77.4 and 97.2%, respectively, and the specificity was 100% for both methods. The results of the Etest AmpC were inconclusive for 10 isolates. With the CC-DDS method 2 inconclusive results were observed. Based on this study, we propose a comprehensive diagnostic flow chart for the detection of AmpC production consisting of a simple phenotypic screening and a single phenotypic confirmation test with inconclusive results being resolved by molecular analysis. For the proposed flow chart using (i) cefoxitin as a screening marker for AmpC production, (ii) the CC-DDS method as phenotypic confirmation, and (iii) molecular methods in case of inconclusive results, the sensitivity and specificity for AmpC detection would have been 97.4 and 100%, respectively, with respect to the studied isolates. The phenotypic methods used in the AmpC algorithm are simple to perform and easy to implement in the diagnostic laboratory. PMID- 21632896 TI - Travel-related Streptococcal toxic shock syndrome caused by emm type 78 Streptococcus pyogenes. AB - Streptococcal toxic shock syndrome is a serious health problem in developed and developing countries. We here report a case of severe protracted disease after a minor skin infection in a young traveler returning from West Malaysia which was caused by an unusual emm-type strain harboring speG and smeZ superantigen genes. PMID- 21632897 TI - Evaluation of two molecular assays for rapid detection of mycobacterium tuberculosis resistance to fluoroquinolones in high-tuberculosis and -multidrug resistance Settings. AB - The Russian Federation is a high-tuberculosis (TB)-burden country with high rates of drug resistance, including multidrug and extensive drug resistance to TB (M/XDRTB). Rapid diagnosis of resistance to fluoroquinolones (FQs) using molecular assays is essential for the implementation of appropriate drug regimens and prevention of the transmission of XDR strains. A total of 51 individual MDRTB strains were tested by pyrosequencing of the quinolone resistance determining region of the gyrA gene and the GenoType MTBDRsl assay (Hain Lifescience, GmbH, Nehren, Germany), and the results were evaluated against those obtained by phenotypic drug susceptibility testing (DST). Mutations were detected in 25 (78.1%) FQ-resistant strains, with the majority of mutations (n = 19 [73.0%]) found in codon 94 of the gyrA gene; the novel mutation 1457 C->Tau was found in the gyrB gene. Three mixed allelic variants were detected, which is a well-known phenomenon in areas with high TB and drug-resistant TB rates. The sensitivity and specificity of pyrosequencing (86.2 and 100%, respectively) and MTBDRsl (86.2 and 100%, respectively) were high; however, the results for 5.9% of the analyzed strains were unreadable when MTBDRsl was used. The MTBDRsl and pyrosequencing assays offer a rapid and accurate means for diagnosing resistance to FQs in high TB-burden areas. PMID- 21632898 TI - Clonal evolution leading to maintenance of antibiotic resistance rates among colonizing Pneumococci in the PCV7 era in Portugal. AB - The introduction of the seven-valent pneumococcal conjugate vaccine (PCV7) in Portugal led to extensive serotype replacement among carriers of pneumococci, with a marked decrease of PCV7 types. Although antimicrobial resistance was traditionally associated with PCV7 types, no significant changes in the rates of nonsusceptibility to penicillin, resistance to macrolides, or multidrug resistance were observed. This study aimed to investigate the mechanisms leading to maintenance of antimicrobial resistance, despite marked serotype replacement. We compared, through molecular typing, 252 antibiotic-resistant pneumococci recovered from young carriers in 2006 and 2007 (era of high PCV7 uptake) with collections of isolates from 2002 and 2003 (n=374; low-PCV7-uptake era) and 1996 to 2001 (n=805; pre-PCV7 era). We observed that the group of clones that has accounted for antimicrobial resistance since 1996 is essentially the same as the one identified in the PCV7 era. The relative proportions of such clones have, however, evolved substantially overtime. Notably, widespread use of PCV7 led to an expansion of two Pneumococcal Molecular Epidemiology Network (PMEN) clones expressing non-PCV7 capsular variants of the original strains: Sweden(15A)ST63 (serotypes 15A and 19A) and Denmark(14)ST230 (serotypes 19A and 24F). These variants were already in circulation in the pre-PCV7 era, although they have now become increasingly abundant. Emergence of novel clones and de novo acquisition of resistance contributed little to the observed scenario. No evidence of capsular switch events occurring after PCV7 introduction was found. In the era of PCVs, antimicrobial resistance remains a problem among the carried pneumococci. Continuous surveillance is warranted to evaluate serotype and clonal shifts leading to maintenance of antimicrobial resistance. PMID- 21632899 TI - Unusual genotype of a Uropathogenic Escherichia coli strain assigned to the B2 phylogenetic group. PMID- 21632900 TI - Bacteremia with Bacteroides pyogenes after a cat bite. AB - Animal bite wounds are often infected with bacteria from the animal's oral flora. We report what we believe to be the first case of bacteremia with Bacteroides pyogenes resulting from an infected cat bite. PMID- 21632901 TI - Comparative evaluation of combined-disk tests using different boronic acid compounds for detection of klebsiella pneumoniae carbapenemase-producing enterobacteriaceae clinical isolates. AB - The accurate phenotypic detection of Klebsiella pneumoniae carbapenemase (KPC) producing Enterobacteriaceae is an increasing necessity worldwide. We evaluated the performance of boronic acid combined-disk tests using as substrate imipenem or meropenem and as inhibitor of KPC production 300 MUg aminophenylboronic acid (APBA), 600 MUg APBA, or 400 MUg phenylboronic acid (PBA). Tests were considered positive when an increase in the growth-inhibitory zone around a carbapenem disk with KPC inhibitor was 5 mm or greater of the growth-inhibitory zone diameter around the disk containing carbapenem alone. The comparison of the combined-disk tests was performed with 112 genotypically confirmed KPC-possessing Enterobacteriaceae isolates. To measure the specificity of the tests, 127 genotypically confirmed KPC-negative Enterobacteriaceae isolates that were nonsusceptible to at least one carbapenem were chosen for testing. Using disks containing imipenem without and with 300 MUg APBA, 600 MUg APBA, or 400 MUg PBA, 72, 92, and 112 of the KPC producers, respectively, gave positive results (sensitivities, 64.3%, 82.1%, and 100%, respectively). Using disks containing meropenem without and with 300 MUg APBA, 600 MUg APBA, or 400 MUg PBA, 87, 108, and 112 of the KPC producers, respectively, gave positive results (sensitivities, 77.7%, 96.4%, and 100%, respectively). Among KPC producers, the disk potentiation tests using meropenem and PBA demonstrated the largest differences in inhibition zones (P < 0.001). All combined-disk tests correctly identified 124 of the 127 non-KPC producers (specificity, 97.6%). This comparative study showed that PBA is the most effective inhibitor of KPC enzymes, and its use in combined-disk tests with meropenem may give the most easily interpreted results. PMID- 21632902 TI - Use of modified PCR ribotyping for direct detection of Clostridium difficile ribotypes in stool samples. AB - PCR ribotyping was modified to allow direct detection of Clostridium difficile from stool samples. Direct PCR ribotyping was possible in 86 out of 99 C. difficile-positive stool samples, and in 84 cases (84.8%), the ribotype determined directly from the stool sample was identical to the ribotype of the strain isolated from the same stool sample. PMID- 21632903 TI - Testosterone or 17{beta}-estradiol exposure reveals sex-specific effects on glucose and lipid metabolism in human myotubes. AB - Changes in sex hormone levels with aging or illness may lead to metabolic disorders. Moreover, the ratio changes in men versus women may have distinct pathological responses. Since little is known about sex hormone action on muscle metabolism, we examined the role of testosterone or 17beta-estradiol (E(2)) in metabolism and investigated whether either hormone may mediate a sex-specific effect. Myotubes from postmenopausal women and age-matched male donors were treated with 10 nM testosterone or E(2) for 4 days, and assays were performed to measure metabolic readouts, signal transduction, and mRNA expression. Testosterone and E(2) treatment enhanced insulin-stimulated glucose incorporation into glycogen and AKT phosphorylation in myotubes from female donors, highlighting a sex-specific role of sex hormone in glucose metabolism. Testosterone treatment increased palmitate oxidation in myotubes from both female and male donors, while E(2) enhanced palmitate oxidation in myotubes from male donors only. Testosterone-mediated increase in palmitate oxidation was attenuated at the presence of androgen receptor antagonist, which may indicate a role of nuclear steroid receptor in muscle lipid oxidation. Testosterone treatment increased mRNA expression of the insulin receptor substrate 2 in myotubes from male and female donors, whereas it increased mRNA expression of glycogen synthase 1 only in myotubes from male donors. E(2) treatment increased pyruvate dehydrogenase kinase 4 mRNA expression in myotubes from female donors. Thus, our data suggest that testosterone or E(2) modulates muscle glucose and lipid metabolism and may play a role in metabolism in a sex-dependent manner. PMID- 21632904 TI - Yapsin 1 immunoreactivity in {alpha}-cells of human pancreatic islets: implications for the processing of human proglucagon by mammalian aspartic proteases. AB - Yapsin 1 is an aspartic protease from Saccharomyces cerevisiae and belongs to a class of aspartic proteases that demonstrate specificity for basic amino acids. It is capable of processing prohormone substrates at specific basic residue cleavage sites, similar to that of the prohormone convertases, to generate bioactive peptide hormones. An antibody raised against yapsin 1 was previously shown to immunostain endocrine cells of rat pituitary and brain as well as lysates from bovine pituitary secretory granules demonstrating the existence of yapsin 1-like aspartic proteases in mammalian endocrine tissues, potentially involved in peptide hormone production. Here, we show the specific staining of yapsin 1 immunoreactivity in the alpha-cells of human pancreatic islets. No staining was observed in the beta- or delta-cells, indicating a specificity of the staining for glucagon-producing and not insulin- or somatostatin-producing cells. Purified yapsin 1 was also shown to process proglucagon into glucagon in vitro, demonstrating that the prototypical enzyme of this subclass of enzymes can correctly process proglucagon to glucagon. These findings suggest the existence of a yapsin 1-like enzyme exclusively in the alpha-cells of the islets of Langerhans in humans, which may play a role in the production of glucagon in that tissue. PMID- 21632905 TI - The pain of cholesterol-lowering therapy. PMID- 21632907 TI - Scar reactivation and dry cough. PMID- 21632906 TI - Immune thrombocytopenia: no longer 'idiopathic'. AB - Immune thrombocytopenia (ITP) is a common hematologic disorder. Its pathogenesis involves both accelerated platelet destruction and impaired platelet production. First-line agents are usually effective initially but do not provide long-term responses. Splenectomy remains an effective long-term therapy, as does rituximab (Rituxan) in a subset of patients. Thrombopoietic agents offer a new alternative, although their place in the overall management of ITP remains uncertain. PMID- 21632908 TI - Subphrenic abscess from a perforated duodenal ulcer. PMID- 21632909 TI - How to manage type 2 diabetes in medical and surgical patients in the hospital. AB - Many patients admitted to the hospital have diabetes mellitus-diagnosed or undiagnosed-and others develop hyperglycemia from the stress of hospitalization. This paper discusses the prevalence, outcomes, and evidence for best management of hyperglycemia and diabetes in hospitalized patients outside the critical care setting. PMID- 21632910 TI - Progressive muscle weakness: More there than meets the eye. PMID- 21632911 TI - Statin myopathy: a common dilemma not reflected in clinical trials. AB - Although statins are remarkably effective, they are still underprescribed because of concerns about muscle toxicity. We review the aspects of statin myopathy that are important to the primary care physician and provide a guide for evaluating patients on statins who present with muscle complaints. We outline the differential diagnosis, the risks and benefits of statin therapy in patients with possible toxicity, and the subsequent treatment options. PMID- 21632912 TI - ST-segment depression and T-wave inversion: classification, differential diagnosis, and caveats. AB - Heightened awareness of the characteristic patterns of ST-segment depression and T-wave inversion is paramount to quickly identifying life-threatening disorders. This paper reviews how to distinguish the various causes of these abnormalities. PMID- 21632913 TI - Development of the HD-Teen Inventory. AB - Adolescents, who have a parent with Huntington Disease (HD), not only are at genetic risk for HD but also are witness to its onset and devastating clinical progression as their parent declines. To date, no mechanism has been developed to direct health care providers to the atypical adolescent experiences of these teens. The purpose of this report is to describe the process of developing the HD Teen Inventory clinical assessment tool. Forty-eight teens and young adults from 19 U.S. states participated in the evaluation of the HD-Teen Inventory tool. Following item analysis, the number of items was reduced and item frequency and reaction scales were combined, based on the strong correlation (r = .94). The resultant tool contains 15 inventory and 2 open-ended response items. The HD-Teen Inventory emerged as a more compact and efficient tool for identifying the most salient concerns of at-risk teens in HD families in research and/or clinical practice. PMID- 21632914 TI - Recognition and management of the latex-allergic patient in the ambulatory plastic surgical suite. AB - Allergic (IgE antibody-mediated) reactions to proteins retained in natural rubber latex (NRL), although decreasing in prevalence, are still a concern in many clinical practices due to possible sensitization of healthcare workers exposed to latex in the office setting and patients with histories of latex allergy who present in the office. In this article, the authors outline the risk factors for latex allergy, describe the symptoms of latex allergy, and provide direction on how to create an environment safe for medical care of latex-allergic patients. In addition, treatment of anaphylaxis to NRL is outlined, including precrisis steps in preparation for anaphylactic reactions in the office surgery setting. Many of the precautions outlined in this article could be extrapolated for a healthcare worker with known or suspected latex allergy. PMID- 21632915 TI - Incidence of breast and chest wall asymmetries: 4D photography. AB - BACKGROUND: Few authors have addressed preoperative soft tissue and chest wall analysis as it pertains to asymmetries that must be identified for preoperative breast augmentation planning. OBJECTIVES: The authors evaluate the incidence of breast and chest wall asymmetries. METHODS: In the present study, 125 consecutive patients underwent a voluntary dimensional analysis by a single practitioner, followed by a confirmatory 4D photographic analysis. During each patient's evaluation, the following measurements were recorded: distance from nipple to inframammary fold (IMF), base width, distance from sternal notch to nipple, horizontal areolar width, vertical areolar height, upper and lower pole pinch tests, and medial and lateral pinch tests. RESULTS: Following exclusion of patients who had undergone prior breast surgery, 117 patients were included in the final statistical analysis. Significant differences between right and left breasts were found in 81.7% of patients in one or more of the measured dimensions (p < .05). The manual measurements were confirmed with computerized 4D photography, and there was no significant difference found between the two measurement types in any objective parameter. However, there was a significant difference in the level of chest wall asymmetries identified by 4D photography. Nipple-to-IMF position asymmetry was present in 59.6% of the patients, and sternal notch-to-nipple asymmetry was present in 81.2%. Overall, 100% of the women had some degree of asymmetry (soft tissue and/or chest wall) confirmed by 4D photography. CONCLUSIONS: The 4D photography measurements in this study were consistent with objective manual measurements but provided the added benefit of identifying chest wall asymmetries more objectively. The data from this study underscore the importance of developing a systematic preoperative breast and chest wall analysis that can be individualized for each patient. The resulting asymmetries should then be discussed with the patient, along with the potential for continued or more pronounced asymmetry postoperatively. PMID- 21632916 TI - Complexities of Rett syndrome and MeCP2. PMID- 21632918 TI - Long-term imaging reveals dynamic changes in the neuronal composition of the glomerular layer. AB - The mammalian olfactory bulb (OB) contains a rich and highly heterogeneous network of local interneurons (INs). These INs undergo continuous turnover in the adult OB in a process known as "adult neurogenesis." Although the overall magnitude of adult neurogenesis has been estimated, the detailed dynamics of the different subpopulations remains largely unknown. Here we present a novel preparation that enables long-term in vivo time-lapse imaging in the mouse OB through a chronic cranial window in a virtually unlimited number of sessions. Using this preparation, we followed the turnover of a specific neuronal population in the OB, the dopaminergic (DA) neurons, for as long as 9 months. By following the same population over long periods of time, we found clear addition and loss of DA neurons in the glomerular layer. Both cell addition and loss increased over time. The numbers of new DA cells were consistently and significantly higher than lost DA cells, suggesting a net increase in the size of this particular population with age. Over a 9 month period of adult life, the net addition of DA neurons reached ~ 13%. Our data argue that the fine composition of the bulbar IN network changes throughout adulthood rather than simply being replenished. PMID- 21632917 TI - Social bonding decreases the rewarding properties of amphetamine through a dopamine D1 receptor-mediated mechanism. AB - Although the protective effects of social bonds on drug use/abuse have been well documented, we know little about the underlying neural mechanisms. Using the prairie vole (Microtus ochrogaster)--a socially monogamous rodent that forms long term pair bonds after mating--we demonstrate that amphetamine (AMPH) conditioning induced a conditioned place preference (CPP) in sexually naive (SN), but not pair bonded (PB), males. Although AMPH treatment induced a similar magnitude of dopamine release in the nucleus accumbens (NAcc) of SN and PB males, it had differential effects on NAcc D1 receptor (D1R) binding. Specifically, AMPH treatment increased D1R binding in SN, but decreased D1R binding in PB males. NAcc D1R, but not D2 receptor, antagonism blocked AMPH-induced CPP in SN males and NAcc D1R activation before AMPH conditioning enabled AMPH-induced CPP in PB males. Together, our data demonstrate that pair-bonding experience decreases the rewarding properties of AMPH through a D1R-mediated mechanism. PMID- 21632919 TI - Short-term facilitation modulates size and timing of the synaptic response at the inner hair cell ribbon synapse. AB - Inner hair cells (IHCs) in the mammalian cochlea are able to continuously release neurotransmitter in the presence of constant stimuli. Nonetheless, strong synaptic depression is observed over the first few milliseconds of stimulation. This process most likely underlies adaptation in the auditory nerve. In the present study we demonstrate that under certain conditions of stimulation, facilitation can occur at the IHC ribbon synapse. Using simultaneous whole-cell, voltage-clamp recordings from IHCs and afferent fiber endings in excised postnatal rat cochleae, we stimulated IHCs with 2 ms long test depolarizations from a holding potential of -89 mV. Synaptic currents in afferent fibers occurred with high failure rates of ~ 50%. However, when a pre-depolarization to values of -55 to -49 mV was implemented before the test pulse, success rates of the synaptic response increased to 100%, the strength of the synaptic response increased ~ 2.8-fold, and synaptic latency was reduced by ~ 50%. When calcium influx was minimized during pre-depolarization, none of these effects were found, suggesting that calcium influx during pre-depolarizations is required for synaptic conditioning. Similarly, in response to paired-pulse protocols, short term facilitation occurred. The response to the second stimulus increased up to ~ 5-fold, and its latency was reduced by up to 35% compared to the response to the first stimulus. We propose that at the IHC resting membrane potential, the ribbon synapse operates in a constantly facilitated mode caused by Ca(2+) influx, optimizing the size and timing of the postsynaptic response in auditory nerve fibers. PMID- 21632920 TI - The cross-modal spread of attention reveals differential constraints for the temporal and spatial linking of visual and auditory stimulus events. AB - The integration of multisensory information has been shown to be guided by spatial and temporal proximity, as well as to be influenced by attention. Here we used neural measures of the multisensory spread of attention to investigate the spatial and temporal linking of synchronous versus near-synchronous auditory and visual events. Human participants attended selectively to one of two lateralized visual-stimulus streams while task-irrelevant tones were presented centrally. Electrophysiological measures of brain activity showed that tones occurring simultaneously or delayed by 100 ms were temporally linked to an attended visual stimulus, as reflected by robust cross-modal spreading-of-attention activity, but not when delayed by 300 ms. The neural data also indicated a ventriloquist-like spatial linking of the auditory to the attended visual stimuli, but only when occurring simultaneously. These neurophysiological results thus provide unique insight into the temporal and spatial principles of multisensory feature integration and the fundamental role attention plays in such integration. PMID- 21632921 TI - Role of guanylyl cyclase modulation in mouse cone phototransduction. AB - A negative phototransduction feedback in rods and cones is critical for the timely termination of their light responses and for extending their function to a wide range of light intensities. The calcium feedback mechanisms that modulate phototransduction in rods have been studied extensively. However, the corresponding modulation mechanisms that enable cones to terminate rapidly their light responses and to adapt in bright light, properties critical for our daytime vision, are still not understood. In cones, calcium feedback to guanylyl cyclase is potentially a key step in phototransduction modulation. The guanylyl cyclase activity is modulated by the calcium-binding guanylyl cyclase activating proteins (GCAP1 and GCAP2). Here, we used single-cell and transretinal recordings from mouse to determine how GCAPs modulate dark-adapted responses as well as light adaptation in mammalian cones. Deletion of GCAPs increased threefold the amplitude and dramatically prolonged the light responses in dark-adapted mouse cones. It also reduced the operating range of mouse cones in background illumination and severely impaired their light adaptation. Thus, GCAPs exert powerful modulation on the mammalian cone phototransduction cascade and play an important role in setting the functional properties of cones in darkness and during light adaptation. Surprisingly, despite their better adaptation capacity and wider calcium dynamic range, mammalian cones were modulated by GCAPs to a lesser extent than mammalian rods. We conclude that a disparity in the strength of GCAP modulation cannot explain the differences in the dark-adapted properties or in the operating ranges of mammalian rods and cones. PMID- 21632922 TI - Sparse but highly efficient Kv3 outpace BKCa channels in action potential repolarization at hippocampal mossy fiber boutons. AB - Presynaptic elements of axons, in which action potentials (APs) cause release of neurotransmitter, are sites of high densities and complex interactions of proteins. We report that the presence of K(v)3 channels in addition to K(v)1 at glutamatergic mossy fiber boutons (MFBs) in rat hippocampal slices considerably limits the number of fast, voltage-activated potassium channels necessary to achieve basal presynaptic AP repolarization. The ~ 10-fold higher repolarization efficacy per K(v)3 channel compared with presynaptic K(v)1 results from a higher steady-state availability at rest, a better recruitment by the presynaptic AP as a result of faster activation kinetics, and a larger single-channel conductance. Large-conductance calcium- and voltage-activated potassium channels (BK(Ca)) at MFBs give rise to a fast activating/fast inactivating and a slowly activating/sustained K(+) current component during long depolarizations. However, BK(Ca) contribute to MFB-AP repolarization only after presynaptic K(v)3 have been disabled. The calcium chelators EGTA and BAPTA are equally effective in preventing BK(Ca) activation, suggesting that BK(Ca) are not organized in nanodomain complexes with presynaptic voltage-gated calcium channels. Thus, the functional properties of K(v)3 channels at MFBs are tuned to both promote brevity of presynaptic APs limiting glutamate release and at the same time keep surface protein density of potassium channels low. Presynaptic BK(Ca) channels are restricted to limit additional increases of the AP half-duration in case of K(v)3 hypofunction, because rapid membrane repolarization by K(v)3 combined with distant calcium sources prevent BK(Ca) activation during basal APs. PMID- 21632923 TI - The cytoskeletal adaptor protein band 4.1B is required for the maintenance of paranodal axoglial septate junctions in myelinated axons. AB - Precise targeting and maintenance of axonal domains in myelinated axons is essential for saltatory conduction. Caspr and Caspr2, which localize at paranodal and juxtaparanodal domains, contain binding sites for the cytoskeletal adaptor protein 4.1B. The exact role of 4.1B in the organization and maintenance of axonal domains is still not clear. Here, we report the generation and characterization of 4.1B-null mice. We show that loss of 4.1B in the PNS results in mislocalization of Caspr at paranodes and destabilization of paranodal axoglial septate junctions (AGSJs) as early as postnatal day 30. In the CNS, Caspr localization is progressively disrupted and ultrastructural analysis showed paranodal regions that were completely devoid of AGSJs, with axolemma separated from the myelin loops, and loops coming off the axolemma. Most importantly, our phenotypic analysis of previously generated 4.1B mutants, used in the study by Horresh et al. (2010), showed that Caspr localization was not affected in the PNS, even after 1 year; and 4.1R was neither expressed, nor enriched at the paranodes. Furthermore, ultrastructural analysis of these 4.1B mutants showed destabilization of CNS AGSJs at ~ 1 year. We also discovered that the 4.1B locus is differentially expressed in the PNS and CNS, and generates multiple splice isoforms in the PNS, suggesting 4.1B may function differently in the PNS versus CNS. Together, our studies provide direct evidence that 4.1B plays a pivotal role in interactions between the paranodal AGSJs and axonal cytoskeleton, and that 4.1B is critically required for long-term maintenance of axonal domains in myelinated axons. PMID- 21632925 TI - Corticospinal beta-range coherence is highly dependent on the pre-stationary motor state. AB - During steady muscle contractions, the human sensorimotor cortex generates oscillations in the beta-frequency range (15-30 Hz) that are coherent with the activity of contralateral spinal motoneurons. This corticospinal coherence is thought to favor stationary motor states, but its mode of operation remains elusive. We hypothesized that corticospinal beta-range coherence depends on the sensorimotor processing state before a steady force task and may thus increase after sensorimotor tuning to dynamic force generation. To test this hypothesis we instructed 16 human subjects to compensate static force after rest as well as after compensating predictable or unpredictable dynamic force with their right index finger. We calculated EEG-EMG coherence, cortical motor spectral power, and the motor performance during the force conditions. Corticospinal beta-coherence during stationary force was excessively elevated if the steady-state contraction was preceded by predictable dynamic force instead of rest, and was highest after unpredictable dynamic force. The beta-power decreased from rest to predictable dynamic force, and was lowest during unpredictable dynamic force. The increase in corticospinal beta-coherence showed a significant negative correlation with the preceding change in beta-power. The tuning to dynamic force did not entail an inferior motor performance during static force. The results imply a correlation between corticospinal beta-range coherence and the computational load of the preceding isometric motor engagement. We suggest beta-range coherence provides a functional corticospinal gateway for steady force-related processing that can override cortical states tuned to dynamic force. The modulation of corticospinal beta-range coherence might thus ensure comparable precision of static force in various motor contexts. PMID- 21632924 TI - Visual experience-dependent maturation of correlated neuronal activity patterns in a developing visual system. AB - The functional properties of neural circuits become increasingly robust over development. This allows circuits to optimize their output in response to a variety of input. However, it is not clear whether this optimization is a function of hardwired circuit elements, or whether it requires neural experience to develop. We performed rapid in vivo imaging of calcium signals from bulk labeled neurons in the Xenopus laevis optic tectum to resolve the rapid spatiotemporal response properties of populations of developing tectal neurons in response to visual stimuli. We found that during a critical time in tectal development, network activity becomes increasingly robust, more correlated, and more synchronous. These developmental changes require normal visual input during development and are disrupted by NMDAR blockade. Our data show that visual activity and NMDAR activation are critical for the maturation of tectal network dynamics during visual system development. PMID- 21632926 TI - Canonical Notch signaling is not necessary for prosensory induction in the mouse cochlea: insights from a conditional mutant of RBPjkappa. AB - The mammalian organ of Corti consists of a highly organized array of hair cells and supporting cells that originate from a common population of prosensory progenitors. Proper differentiation of this complex cellular mosaic requires lateral inhibition mediated by Notch signaling. Several studies have implicated Notch signaling in the earlier induction of the prosensory domain that lies along the length of the cochlear duct, and which forms before the onset of hair cell and supporting cell differentiation. To investigate the role of Notch signaling in prosensory domain formation, we conditionally inactivated the transcriptional mediator of canonical Notch signaling, RBPjkappa, throughout the inner ear. Although RBPjkappa mutants have severe vestibular defects and a shortened cochlear duct, markers of the prosensory domain appear at the normal time and location in the cochlea of RBPjkappa mutants. Despite the lack of RBPjkappa, hair cell and supporting cell markers also appear at appropriate times in the cochlea, suggesting that RBPjkappa is dispensable for differentiation of the cochlear sensory epithelium. However, we also observed that differentiating hair cells and supporting cells rapidly die in RBPjkappa mutants, suggesting a requirement of RBPjkappa for cell survival in this tissue. Finally, in contrast to the chick basilar papilla, ectopic activation of Notch signaling did not induce ectopic sensory patches in nonsensory regions of the cochlea. Our results indicate that canonical Notch signaling is not necessary for prosensory specification in the mouse cochlea, suggesting that other signaling pathways may specify this highly derived sensory organ. PMID- 21632927 TI - Superior colliculus inactivation alters the weighted integration of visual stimuli. AB - The primate superior colliculus (SC) is important for the winner-take-all selection of targets for orienting movements. Such selection takes time, however, and the earliest motor responses typically are guided by a weighted vector average of the visual stimuli, before the winner-take-all selection of a single target. We tested whether SC activity plays a role in this initial stage of orienting by inactivating the SC in two macaques (Macaca mulatta) with local muscimol injections. After SC inactivation, initial orienting responses still followed a vector average, but the contribution of the visual stimulus inside the affected field was decreased, and the contribution of the stimulus outside the affected field was increased. These results demonstrate that the SC plays an important role in the weighted integration of visual signals for orienting, in addition to its role in the winner-take-all selection of the target. PMID- 21632928 TI - G-protein betagamma-complex is crucial for efficient signal amplification in vision. AB - A fundamental question of cell signaling biology is how faint external signals produce robust physiological responses. One universal mechanism relies on signal amplification via intracellular cascades mediated by heterotrimeric G-proteins. This high amplification system allows retinal rod photoreceptors to detect single photons of light. Although much is now known about the role of the alpha-subunit of the rod-specific G-protein transducin in phototransduction, the physiological function of the auxiliary betagamma-complex in this process remains a mystery. Here, we show that elimination of the transducin gamma-subunit drastically reduces signal amplification in intact mouse rods. The consequence is a striking decline in rod visual sensitivity and severe impairment of nocturnal vision. Our findings demonstrate that transducin betagamma-complex controls signal amplification of the rod phototransduction cascade and is critical for the ability of rod photoreceptors to function in low light conditions. PMID- 21632929 TI - Neto2 modulation of kainate receptors with different subunit compositions. AB - Kainate receptors are less well understood than other glutamate receptors, and synaptic kainate receptors display properties that differ from recombinant receptors. In particular, the slow decay of kainate receptor synaptic currents contrasts with the rapid deactivation and desensitization of receptors expressed in heterologous cells. We recently identified Neuropilin and Tolloid like-2 (Neto2) as a novel accessory subunit of kainate receptors and showed that Neto2 modulates the gating kinetics of GluK2 receptors. However, the kainate receptor family consists of five different subunits (GluK1-5) that can form homomeric and heteromeric receptors with different functional properties. Here, we tested whether Neto2 modulation varies with subunit composition. Rapid application techniques were used to apply glutamate to outside-out patches that contained GluK1, GluK1/5, or GluK2/5 kainate receptors. Coexpression of Neto2 slowed desensitization to varying degrees. Responses to 1 ms pulses of glutamate were also slowed by Neto2, especially for receptors containing GluK5, as were postsynaptic currents in neurons expressing recombinant kainate receptors. In addition, Neto2 markedly increased the rate at which some receptors recovered from desensitization. These results suggest that Neto2 modulates the function of most kainate receptors. PMID- 21632931 TI - Spinning versus wobbling: how the brain solves a geometry problem. AB - Oscillating an animal out-of-phase simultaneously about the roll and pitch axes ("wobble") changes continuously the orientation of the head relative to gravity. For example, it may gradually change from nose-up, to ear-down, nose-down, ear down, and back to nose-up. Rotations about the longitudinal axis ("spin") can change the orientation of the head relative to gravity in the same way, provided the axis is tilted from vertical. During both maneuvers, the otolith organs in the inner ear detect the change in head orientation relative to gravity, whereas the semicircular canals will only detect oscillations in velocity (wobble), but not any rotation at constant velocity (spin). Geometrically, the whole motion can be computed based on information about head orientation relative to gravity and the wobble velocity. We subjected monkeys (Macaca mulatta) to combinations of spin and wobble and found that the animals were always able to correctly estimate their spin velocity. Simulations of these results with an optimal Bayesian model of vestibular information processing suggest that the brain integrates gravity and velocity information based on a geometrically coherent three-dimensional representation of head-in-space motion. PMID- 21632930 TI - Phosphoinositide 3-kinase enhancer regulates neuronal dendritogenesis and survival in neocortex. AB - Phosphoinositide 3-kinase enhancer (PIKE) binds and enhances phosphatidylinositol 3-kinase (PI3K)/Akt activities. However, its physiological functions in brain have never been explored. Here we show that PIKE is important in regulating the neuronal survival and development of neocortex. During development, enhanced apoptosis is observed in the ventricular zone of PIKE knock-out (PIKE(-/-)) cortex. Moreover, PIKE(-/-) neurons show reduced dendritic complexity, dendritic branch length, and soma size. These defects are due to the reduced PI3K/Akt activities in PIKE(-/-) neurons, as the impaired dendritic arborization can be rescued when PI3K/Akt cascade is augmented in vitro or in PIKE(-/-)PTEN(-/-) double-knock-out mice. Interestingly, PIKE(-/-) mice display behavioral abnormality in locomotion and spatial navigation. Because of the diminished PI3K/Akt activities, PIKE(-/-) neurons are more vulnerable to glutamate- or stroke-induced neuronal cell death. Together, our data established the critical role of PIKE in regulating neuronal survival and development by substantiating the PI3K/Akt pathway. PMID- 21632932 TI - Apraxia impairs intentional retrieval of incidentally acquired motor knowledge. AB - Apraxia caused by left hemispheric stroke typically impairs skilled sequential movements. After stroke, apraxic patients need to reacquire motor skills by motor learning. The current study assessed for the first time incidental motor sequence learning in apraxic patients. Forty-eight human subjects (henceforth called "patients") with left hemispheric stroke affecting the middle cerebral artery territory (18 with apraxia and 30 without apraxia) and 17 age-matched healthy controls were tested on a visuomanual serial reaction time task. Subjects performed four blocks consisting of repetitions of a complex six element sequence containing ambiguous pairwise transitions before a new and unfamiliar sequence was introduced in block 5. Reaction time (RT) disadvantages in this fifth block indicated incidental sequence-specific motor learning. The intentional retrieval of the learned motor knowledge was assessed subsequently with a free recall task. Voxel-based lesion-symptom mapping (VLSM) was performed to investigate for the first time the lesion correlates of deficits in learning and retrieving sequential motor knowledge. Despite generally prolonged RTs, apraxic patients showed sequence-specific motor learning as could be observed in nonapraxic patients and healthy controls. However, apraxic patients showed reduced intentional retrieval of the learned sequence. VLSM revealed that impaired intentional retrieval of motor sequence knowledge resulted from dorsal premotor cortex lesions. Apraxic patients showed a dissociation of preserved incidental motor (sequence) learning and deficient intentional retrieval of this incidentally learned motor knowledge. The data suggest that novel approaches for treating apraxia should focus on incidental motor learning, but that automatic rather than intentional retrieval strategies should be enforced. PMID- 21632933 TI - Girdin is an intrinsic regulator of neuroblast chain migration in the rostral migratory stream of the postnatal brain. AB - In postnatally developing and adult brains, interneurons of the olfactory bulb (OB) are continuously generated at the subventricular zone of the forebrain. The newborn neuroblasts migrate tangentially to the OB through a well defined pathway, the rostral migratory stream (RMS), where the neuroblasts undergo collective migration termed "chain migration." The cell-intrinsic regulatory mechanism of neuroblast chain migration, however, has not been uncovered. Here we show that mice lacking the actin-binding Akt substrate Girdin (a protein that interacts with Disrupted-In-Schizophrenia 1 to regulate neurogenesis in the dentate gyrus) have profound defects in neuroblast chain migration along the RMS. Analysis of two gene knock-in mice harboring Girdin mutants identified unique amino acid residues in Girdin's C-terminal domain that are responsible for the regulation of neuroblast chain migration but revealed no apparent requirement of Girdin phosphorylation by Akt. Electron microscopic analyses demonstrated the involvement of Girdin in neuroblast cell-cell interactions. These findings suggest that Girdin is an important intrinsic factor that specifically governs neuroblast chain migration along the RMS. PMID- 21632934 TI - Olfactory cortex generates synchronized top-down inputs to the olfactory bulb during slow-wave sleep. AB - The olfactory cortex is functionally isolated from the external odor world during slow-wave sleep. However, the neuronal activity pattern in the olfactory cortex and its functional roles during slow-wave sleep are not well understood. Here, we demonstrate in freely behaving rats that the anterior piriform cortex, a major area of the olfactory cortex, repeatedly generates sharp waves that are accompanied by synchronized discharges of numerous cortical neurons. Olfactory cortex sharp waves occurred relatively independently of hippocampal sharp waves. Current source density analysis showed that sharp wave generation involved the participation of recurrent association fiber synapses to pyramidal cells in the olfactory cortex. During slow-wave sleep, the olfactory bulb showed sharp waves that were in synchrony with olfactory cortex sharp waves, indicating that olfactory cortex sharp waves drove synchronized top-down inputs to the olfactory bulb. Based on these results, we speculate that the olfactory cortex sharp waves may play a role in the reorganization of bulbar neuronal circuits during slow wave sleep. PMID- 21632936 TI - Dorsal and ventral parietal contributions to spatial orienting in the human brain. AB - Influential functional magnetic resonance imaging (fMRI)-based models have involved a dorsal frontoparietal network in the orienting of both endogenous and exogenous attention, and a ventral system in attentional reorienting to task relevant events. Nonetheless, given the low temporal resolution and susceptibility to epiphenomenal activations of fMRI, such depictions remain highly debated. We hereby benefited from the high temporal resolution and causal power of event-related transcranial magnetic stimulation to explore the implications of key dorsal and ventral parietal regions in those two types of attention. We provide for the first time causal evidence of right intraparietal sulcus involvement in both types of attentional orienting, while we link the temporoparietal junction with the orienting of exogenous but not endogenous spatial attention. PMID- 21632935 TI - Presynaptic alpha2-GABAA receptors in primary afferent depolarization and spinal pain control. AB - Spinal dorsal horn GABA(A) receptors are found both postsynaptically on central neurons and presynaptically on axons and/or terminals of primary sensory neurons, where they mediate primary afferent depolarization (PAD) and presynaptic inhibition. Both phenomena have been studied extensively on a cellular level, but their role in sensory processing in vivo has remained elusive, due to inherent difficulties to selectively interfere with presynaptic receptors. Here, we address the contribution of a major subpopulation of GABA(A) receptors (those containing the alpha2 subunit) to spinal pain control in mice lacking alpha2 GABA(A) receptors specifically in primary nociceptors (sns-alpha2(-/-) mice). sns alpha2(-/-) mice exhibited GABA(A) receptor currents and dorsal root potentials of normal amplitude in vitro, and normal response thresholds to thermal and mechanical stimulation in vivo, and developed normal inflammatory and neuropathic pain sensitization. However, the positive allosteric GABA(A) receptor modulator diazepam (DZP) had almost completely lost its potentiating effect on PAD and presynaptic inhibition in vitro and a major part of its spinal antihyperalgesic action against inflammatory hyperalgesia in vivo. Our results thus show that part of the antihyperalgesic action of spinally applied DZP occurs through facilitated activation of GABA(A) receptors residing on primary nociceptors. PMID- 21632937 TI - Striatal-enriched protein tyrosine phosphatase expression and activity in Huntington's disease: a STEP in the resistance to excitotoxicity. AB - Striatal-enriched protein tyrosine phosphatase (STEP) is highly expressed in striatal projection neurons, the neuronal population most affected in Huntington's disease. Here, we examined STEP expression and phosphorylation, which regulates its activity, in N-terminal exon-1 and full-length mutant huntingtin mouse models. R6/1 mice displayed reduced STEP protein levels in the striatum and cortex, whereas its phosphorylation was increased in the striatum, cortex, and hippocampus. The early increase in striatal STEP phosphorylation levels correlated with a deregulation of the protein kinase A pathway, and decreased calcineurin activity at later stages further contributes to an enhancement of STEP phosphorylation and inactivation. Accordingly, we detected an accumulation of phosphorylated ERK2 and p38, two targets of STEP, in R6/1 mice striatum at advanced stages of the disease. Activation of STEP participates in excitotoxic-induced cell death. Because Huntington's disease mouse models develop resistance to excitotoxicity, we analyzed whether decreased STEP activity was involved in this process. After intrastriatal quinolinic acid (QUIN) injection, we detected higher phosphorylated STEP levels in R6/1 than in wild-type mice, suggesting that STEP inactivation could mediate neuroprotection in R6/1 striatum. In agreement, intrastriatal injection of TAT-STEP increased QUIN-induced cell death. R6/2, Tet/HD94, and Hdh(Q7/Q111) mice striatum also displayed decreased STEP protein and increased phosphorylation levels. In Tet/HD94 mice striatum, mutant huntingtin transgene shutdown reestablished STEP expression. In conclusion, the STEP pathway is severely downregulated in the presence of mutant huntingtin and may participate in compensatory mechanisms activated by striatal neurons that lead to resistance to excitotoxicity. PMID- 21632939 TI - Brain modularity in arthropods: individual neurons that support "what" but not "where" memories. AB - Experiments with insects and crabs have demonstrated their remarkable capacity to learn and memorize complex visual features (Giurfa et al., 2001; Pedreira and Maldonado, 2003; Chittka and Niven, 2009). Such abilities are thought to require modular brain processing similar to that occurring in vertebrates (Menzel and Giurfa, 2001). Yet, physiological evidence for this type of functioning in the small brains of arthropods is still scarce (Liu et al., 1999, 2006; Menzel and Giurfa, 2001). In the crab Chasmagnathus granulatus, the learning rate as well as the long-term memory of a visual stimulus has been found to be reflected in the performance of identified lobula giant neurons (LGs) (Tomsic et al., 2003). The memory can only be evoked in the training context, indicating that animals store two components of the learned experience, one related to the visual stimulus and one related to the visual context (Tomsic et al., 1998; Hermitte et al., 1999). By performing intracellular recordings in the intact animal, we show that the ability of crabs to generalize the learned stimulus into new space positions and to distinguish it from a similar but unlearned stimulus, two of the main attributes of stimulus memory, is reflected by the performance of the LGs. Conversely, we found that LGs do not support the visual context memory component. Our results provide physiological evidence that the memory traces regarding "what" and "where" are stored separately in the arthropod brain. PMID- 21632938 TI - A silent synapse-based mechanism for cocaine-induced locomotor sensitization. AB - Locomotor sensitization is a common and robust behavioral alteration in rodents whereby following exposure to abused drugs such as cocaine, the animal becomes significantly more hyperactive in response to an acute drug challenge. Here, we further analyzed the role of cocaine-induced silent synapses in the nucleus accumbens (NAc) shell and their contribution to the development of locomotor sensitization. Using a combination of viral vector-mediated genetic manipulations, biochemistry, and electrophysiology in a locomotor sensitization paradigm with repeated, daily, noncontingent cocaine (15 mg/kg) injections, we show that dominant-negative cAMP-element binding protein (CREB) prevents cocaine induced generation of silent synapses of young (30 d old) rats, whereas constitutively active CREB is sufficient to increase the number of NR2B containing NMDA receptors (NMDARs) at synapses and to generate silent synapses. We further show that occupancy of CREB at the NR2B promoter increases and is causally related to the increase in synaptic NR2B levels. Blockade of NR2B containing NMDARs by administration of the NR2B-selective antagonist Ro256981 directly into the NAc, under conditions that inhibit cocaine-induced silent synapses, prevents the development of cocaine-elicited locomotor sensitization. Our data are consistent with a cellular cascade whereby cocaine-induced activation of CREB promotes CREB-dependent transcription of NR2B and synaptic incorporation of NR2B-containing NMDARs, which generates new silent synapses within the NAc. We propose that cocaine-induced activation of CREB and generation of new silent synapses may serve as key cellular events mediating cocaine-induced locomotor sensitization. These findings provide a novel cellular mechanism that may contribute to cocaine-induced behavioral alterations. PMID- 21632940 TI - Synchronized bilateral synaptic inputs to Drosophila melanogaster neuropeptidergic rest/arousal neurons. AB - Neuropeptide PDF (pigment-dispersing factor)-secreting large ventrolateral neurons (lLN(v)s) in the Drosophila brain regulate daily patterns of rest and arousal. These bilateral wake-promoting neurons are light responsive and integrate information from the circadian system, sleep circuits, and light environment. To begin to dissect the synaptic circuitry of the circadian neural network, we performed simultaneous dual whole-cell patch-clamp recordings of pairs of lLN(v)s. Both ipsilateral and contralateral pairs of lLN(v)s exhibit synchronous rhythmic membrane activity with a periodicity of ~ 5-10 s. This rhythmic lLN(v) activity is blocked by TTX, voltage-gated sodium blocker, or alpha-bungarotoxin, nicotinic acetylcholine receptor antagonist, indicating that action potential-dependent cholinergic synaptic connections are required for rhythmic lLN(v) activity. Since injecting current into one neuron of the pair had no effect on the membrane activity of the other neuron of the pair, this suggests that the synchrony is attributable to bilateral inputs and not coupling between the pairs of lLN(v)s. To further elucidate the nature of these synaptic inputs to lLN(v)s, we blocked or activated a variety of neurotransmitter receptors and measured effects on network activity and ionic conductances. These measurements indicate the lLN(v)s possess excitatory nicotinic ACh receptors, inhibitory ionotropic GABA(A) receptors, and inhibitory ionotropic GluCl (glutamate-gated chloride) receptors. We demonstrate that cholinergic input, but not GABAergic input, is required for synchronous membrane activity, whereas GABA can modulate firing patterns. We conclude that neuropeptidergic lLN(v)s that control rest and arousal receive synchronous synaptic inputs mediated by ACh. PMID- 21632941 TI - NMDA receptor activation suppresses microtubule growth and spine entry. AB - Dynamic microtubules are important to maintain neuronal morphology and function, but whether neuronal activity affects the organization of dynamic microtubules is unknown. Here, we show that a protocol to induce NMDA-dependent long-term depression (LTD) rapidly attenuates microtubule dynamics in primary rat hippocampal neurons, removing the microtubule-binding protein EB3 from the growing microtubule plus-ends in dendrites. This effect requires the entry of calcium and is mediated by activation of NR2B-containing NMDA-type glutamate receptor. The rapid NMDA effect is followed by a second, more prolonged response, during which EB3 accumulates along MAP2-positive microtubule bundles in the dendritic shaft. MAP2 is both required and sufficient for this activity-dependent redistribution of EB3. Importantly, NMDA receptor activation suppresses microtubule entry in dendritic spines, whereas overexpression of EB3-GFP prevents NMDA-induced spine shrinkage. These results suggest that short-lasting and long lasting changes in dendritic microtubule dynamics are important determinants for NMDA-induced LTD. PMID- 21632942 TI - Attention improves encoding of task-relevant features in the human visual cortex. AB - When spatial attention is directed toward a particular stimulus, increased activity is commonly observed in corresponding locations of the visual cortex. Does this attentional increase in activity indicate improved processing of all features contained within the attended stimulus, or might spatial attention selectively enhance the features relevant to the observer's task? We used fMRI decoding methods to measure the strength of orientation-selective activity patterns in the human visual cortex while subjects performed either an orientation or contrast discrimination task, involving one of two laterally presented gratings. Greater overall BOLD activation with spatial attention was observed in visual cortical areas V1-V4 for both tasks. However, multivariate pattern analysis revealed that orientation-selective responses were enhanced by attention only when orientation was the task-relevant feature and not when the contrast of the grating had to be attended. In a second experiment, observers discriminated the orientation or color of a specific lateral grating. Here, orientation-selective responses were enhanced in both tasks, but color-selective responses were enhanced only when color was task relevant. In both experiments, task-specific enhancement of feature-selective activity was not confined to the attended stimulus location but instead spread to other locations in the visual field, suggesting the concurrent involvement of a global feature-based attentional mechanism. These results suggest that attention can be remarkably selective in its ability to enhance particular task-relevant features and further reveal that increases in overall BOLD amplitude are not necessarily accompanied by improved processing of stimulus information. PMID- 21632943 TI - Grasping-related functional magnetic resonance imaging brain responses in the macaque monkey. AB - Research in recent decades has suggested the existence of a dedicated brain network devoted to the organization and execution of grasping, one of the most important and skilled movements of primates. Grasping an object requires the transformation of intrinsic object properties such as size, orientation, and shape into an appropriate motor scheme shaping the hand. Although electrophysiological recordings in the monkey model have proven invaluable for gaining insights into the neuronal substrate underlying this complex behavior, knowledge concerning the existence and organization of a similar system in the human brain is derived mainly from imaging studies. Here, we present for the first time functional magnetic resonance imaging (fMRI) of brain activity while macaque monkeys performed reaching and grasping movements in a 3 tesla MR scanner. Grasping in the dark (compared with reaching) yielded significant activations in anterior intraparietal area and ventral premotor area F5, in addition to area PFG in the rostral inferior parietal lobule, somatosensory areas (SI, SII, area 5), and the hand field of F1. Whole-brain macaque fMRI motor studies will be instrumental in establishing possible homologies concerning grasping organization in the human and monkey brains, bridging the gap between human imaging and monkey electrophysiology. PMID- 21632944 TI - Effects of stimulus direction on the correlation between behavior and single units in area MT during a motion detection task. AB - Trial-to-trial variations in the firing rates of neurons in the middle temporal visual area (MT) are correlated with the behavior of macaque monkeys performing motion detection and motion discrimination tasks. Here we examine how these correlations depend on the direction of motion used for a detection task relative to the preferred direction of the neuron under study. There was a robust correlation between the firing rate of MT neurons and the animal's detection of motion when the direction of that motion was within ~ 45 degrees of the preferred direction of the neuron. This correlation was undetectable using motions that were ~ 90 degrees away from the preferred direction, and an inverse correlation between activity and behavior was found for motion in the null direction. Correlation between reaction times and single-cell activity in MT followed a similar pattern. Although motion detection could have been based solely on the activity of neurons preferring the expected direction, these results suggest that it depends on the relative activity of neurons preferring opposite directions of motion. They furthermore show that the subset of neurons used to guide behavior can vary from trial to trial based on task requirements. PMID- 21632946 TI - Scratching beneath the surface: new insights into the functional properties of the lateral occipital area and parahippocampal place area. AB - We used fMRI on neurologically intact humans to investigate whether or not there are different neural substrates for the different kinds of information that a visual surface signals (shape from texture vs material properties from texture). Participants attended to differences in the shape (flat/convex), texture and color (wood/rock), or material properties (soft/hard) of a set of circular surfaces. Attending to shape activated the contour-sensitive lateral occipital (LO) area, and attending to texture activated a region of the collateral sulcus (CoS) that overlaps with the parahippocampal place area (PPA). Interestingly, attending to material properties activated the same texture-sensitive region in the CoS. These results demonstrate the existence of different neural substrates for the different types of information that a visual surface signals. With regard to object shape, the organization of the LO area may be complex, with neurons tuned not only to the outline shape of objects, but also to their surface curvature independent of contour. Moreover, to our knowledge, this is the first study to demonstrate that processing surface texture, which occurs within the scene-sensitive PPA, is a route to accessing knowledge about an object's material properties. With this in mind, we propose that models of visual cortical organization should focus not only on the particular stimulus category to which a region maximally responds (e.g., objects, scenes), but also on the stimulus attributes that best support the processing of that category (e.g., shape, texture, material properties). PMID- 21632945 TI - Multiple routes from occipital to temporal cortices during reading. AB - Contemporary models of the neural system that supports reading propose that activity in a ventral occipitotemporal area (vOT) drives activity in higher-order language areas, for example, those in the posterior superior temporal sulcus (pSTS) and anterior superior temporal sulcus (aSTS). We used fMRI with dynamic causal modeling (DCM) to investigate evidence for other routes from visual cortex to the left temporal lobe language areas. First we identified activations in posterior inferior occipital (iO) and vOT areas that were more activated for silent reading than listening to words and sentences; and in pSTS and aSTS areas that were commonly activated for reading relative to false-fonts and listening to words relative to reversed words. Second, in three different DCM analyses, we tested whether visual processing of words modulates activity from the following: (1) iO->vOT, iO->pSTS, both, or neither; (2) vOT->pSTS, iO->pSTS, both or neither; and (3) pSTS->aSTS, vOT->aSTS, both, or neither. We found that reading words increased connectivity (1) from iO to both pSTS and vOT; (2) to pSTS from both iO and vOT; and (3) to aSTS from both vOT and pSTS. These results highlight three potential processing streams in the occipitotemporal cortex: iO->pSTS >aSTS; iO->vOT->aSTS; and iO->vOT->pSTS->aSTS. We discuss these results in terms of cognitive models of reading and propose that efficient reading relies on the integrity of all these pathways. PMID- 21632947 TI - Cognitive effort drives workspace configuration of human brain functional networks. AB - Effortful cognitive performance is theoretically expected to depend on the formation of a global neuronal workspace. We tested specific predictions of workspace theory, using graph theoretical measures of network topology and physical distance of synchronization, in magnetoencephalographic data recorded from healthy adult volunteers (N = 13) during performance of a working memory task at several levels of difficulty. We found that greater cognitive effort caused emergence of a more globally efficient, less clustered, and less modular network configuration, with more long-distance synchronization between brain regions. This pattern of task-related workspace configuration was more salient in the beta-band (16-32 Hz) and gamma-band (32-63 Hz) networks, compared with both lower (alpha-band; 8-16 Hz) and higher (high gamma-band; 63-125 Hz) frequency intervals. Workspace configuration of beta-band networks was also greater in faster performing participants (with correct response latency less than the sample median) compared with slower performing participants. Processes of workspace formation and relaxation in relation to time-varying demands for cognitive effort could be visualized occurring in the course of task trials lasting <2 s. These experimental results provide support for workspace theory in terms of complex network metrics and directly demonstrate how cognitive effort breaks modularity to make human brain functional networks transiently adopt a more efficient but less economical configuration. PMID- 21632949 TI - Cav1.3 calcium channels are required for normal development of the auditory brainstem. AB - Within the Ca(v)1 family of voltage-gated calcium channels, Ca(v)1.2 and Ca(v)1.3 channels are the predominant subtypes in the brain. Whereas specific functions for each subtype were described in the adult brain, their role in brain development is poorly understood. Here we assess the role of Ca(v)1.3 subunits in the activity-dependent development of the auditory brainstem. We used Ca(v)1.3 deficient (Ca(v)1.3(-/-)) mice because these mice lack cochlea-driven activity that deprives the auditory centers from peripheral input. We found a drastically reduced volume in all auditory brainstem centers (range 25-59%, total 35%), which was manifest before hearing onset. A reduction was not obvious outside the auditory system. The lateral superior olive (LSO) was strikingly malformed in Ca(v)1.3(-/-) mice and had fewer neurons (1/3 less). The remaining LSO neurons displayed normal dendritic trees and received functional glutamatergic input, yet they fired action potentials predominantly with a multiple pattern upon depolarization, in contrast to the single firing pattern prevalent in controls. The latter finding appears to be due to a reduction of dendrototoxin-sensitive potassium conductances, presumably mediated through the K(v)1.2 subtype. Fura2 imaging provided evidence for functional Ca(v)1.3 channels in the LSO of wild type mice. Our results imply that Ca(v)1.3 channels are indispensable for the development of the central auditory system. We propose that the unique LSO phenotype in Ca(v)1.3(-/-) mice, which hitherto was not described in other hereditary deafness models, is caused by the synergistic contribution of two factors: on-site loss of Ca(v)1.3 channels in the neurons plus lack of peripheral input. PMID- 21632948 TI - Protein kinase CK2 increases glutamatergic input in the hypothalamus and sympathetic vasomotor tone in hypertension. AB - Increased glutamatergic input in the paraventricular nucleus (PVN) is important for high sympathetic outflow in hypertension, but the associated molecular mechanisms remain unclear. Here, we determined the role of protein kinase CK2 (formerly casein kinase II) in increased N-methyl-d-aspartate receptor (NMDAR) activity in spinally projecting PVN neurons and sympathetic vasomotor tone in spontaneously hypertensive rats (SHRs). The selective CK2 inhibitors 5,6-dichloro 1-beta-d-ribofuranosylbenzimidazole (DRB) or 4,5,6,7-tetrabromobenzotriazole (TBB) significantly decreased the frequency of miniature EPSCs (mEPSCs) of labeled PVN neurons in SHRs but not in Wistar-Kyoto (WKY) normotensive rats. Also, DRB abolished the inhibitory effect of the NMDAR antagonist AP5 on the frequency of mEPSCs in SHRs. Treatment with DRB or TBB significantly reduced the amplitude of evoked NMDA-EPSCs but not AMPA-EPSCs in SHRs. Furthermore, DRB significantly decreased the firing activity of PVN neurons in SHRs but not in WKY rats. The membrane protein level of CK2alpha in the PVN, but not brainstem and prefrontal cortex, was significantly higher in SHRs than in WKY rats. Lowering blood pressure with celiac ganglionectomy in SHRs did not alter the increased CK2alpha level and the effects of DRB on mEPSCs and NMDA-EPSCs. In addition, intracerebroventricular injection of DRB not only significantly reduced blood pressure and lumbar sympathetic nerve discharges but also eliminated the inhibitory effect of AP5 microinjected into the PVN on sympathetic nerve activity in SHRs. Our findings suggest that augmented CK2 activity critically contributes to increased presynaptic and postsynaptic NMDAR activity in the PVN and elevated sympathetic vasomotor tone in essential hypertension. PMID- 21632950 TI - Suppressive mechanisms in monkey V1 help to solve the stereo correspondence problem. AB - Neurons encode the depth in stereoscopic images by combining the signals from the receptive fields in the two eyes. Local variations in single images can activate neurons that do not signal the correct disparity (false matches), giving rise to the stereo correspondence problem. We used binocular white-noise stimuli to decompose the responses of monkey primary visual cortex V1 neurons into the elements of a linear-nonlinear model (via spike-triggered covariance analysis). In our population of disparity-selective neurons, we find both excitatory and suppressive elements in many of the neurons. Their binocular receptive fields were aligned in a specific push-pull manner for disparity. We demonstrate that this arrangement reduces the responses to false matches but preserves the responses to true matches. The responses of the cells to the noise stimuli were well explained by a linear summation of the elements, followed by a nonlinearity. This model also explained the shape of independently measured disparity-tuning curves, although it overestimated the response magnitude. This study constitutes the first direct physiological evidence for the contribution of suppressive mechanisms to disparity selectivity. This new mechanism contributes to solving the stereo correspondence problem. PMID- 21632952 TI - The benefit of object interactions arises in the lateral occipital cortex independent of attentional modulation from the intraparietal sulcus: a transcranial magnetic stimulation study. AB - Our visual experience is generally not of isolated objects, but of scenes, where multiple objects are interacting. Such interactions (e.g., a watering can positioned to pour water toward a plant) have been shown to facilitate object identification compared with when the objects are depicted as not interacting (e.g., a watering can positioned away from the plant) (Green and Hummel, 2004, 2006). What is the neural basis for this advantage? Recent fMRI studies have identified the lateral occipital cortex (LO) as a potential neural origin of this behavioral benefit, as LO showed greater responses to object pairs depicted as interacting compared with when they are not (Kim and Biederman, 2010; Roberts and Humphreys, 2010). However, it is possible that LO was modulated by an attention sensitive region, the intraparietal sulcus (IPS), which sometimes showed a similar pattern of responses as that of LO in the Kim and Biederman (2010) investigation. To test this hypothesis, we delivered transcranial magnetic stimulation (TMS) to human subjects' LO and IPS while they detected a target object that was or was not interacting with another object to form a scene. TMS delivered to LO but not IPS abolished the facilitation in identifying interacting objects compared with noninteracting depictions observed in the absence of TMS, suggesting that it is LO and not IPS that is critical for the coding of object interactions. PMID- 21632951 TI - Deletion of astroglial Dicer causes non-cell-autonomous neuronal dysfunction and degeneration. AB - The endoribonuclease, Dicer, is indispensable for generating the majority of mature microRNAs (miRNAs), which are posttranscriptional regulators of gene expression involved in a wide range of developmental and pathological processes in the mammalian CNS. Although functions of Dicer-dependent miRNA pathways in neurons and oligodendrocytes have been extensively investigated, little is known about the role of Dicer in astrocytes. Here, we report the effect of Cre-loxP mediated conditional deletion of Dicer selectively from postnatal astroglia on brain development. Dicer-deficient mice exhibited normal motor development and neurological morphology before postnatal week 5. Thereafter, mutant mice invariably developed a rapidly fulminant neurological decline characterized by ataxia, severe progressive cerebellar degeneration, seizures, uncontrollable movements, and premature death by postnatal week 9-10. Integrated transcription profiling, histological, and functional analyses of cerebella showed that deletion of Dicer in cerebellar astrocytes altered the transcriptome of astrocytes to be more similar to an immature or reactive-like state before the onset of neurological symptoms or morphological changes. As a result, critical and mature astrocytic functions including glutamate uptake and antioxidant pathways were substantially impaired, leading to massive apoptosis of cerebellar granule cells and degeneration of Purkinje cells. Collectively, our study demonstrates the critical involvement of Dicer in normal astrocyte maturation and maintenance. Our findings also reveal non-cell-autonomous roles of astrocytic Dicer-dependent pathways in regulating proper neuronal functions and implicate that loss of or dysregulation of astrocytic Dicer-dependent pathways may be involved in neurodegeneration and other neurological disorders. PMID- 21632953 TI - Monitoring the efficacy of a PICU nutrition therapy protocol. PMID- 21632954 TI - Influence of vitamins, trace elements, and iron on lipid peroxidation reactions in all-in-one admixtures for neonatal parenteral nutrition. AB - BACKGROUND: The purpose of this study was to evaluate the effect of vitamins, trace elements, or iron on lipid peroxidation in all-in-one parenteral nutrition (PN) admixtures for preterm neonates. METHODS: Malondialdehyde (MDA) concentrations were analyzed over a 24-hour period (H1-H24) in lipid-containing PN solutions that have a composition identical to that used in the routine clinical care of preterm infants. Six different solutions were prepared and evaluated when exposed to ambient light and light-protected conditions as follows: control (without vitamins [Vit], trace elements [TE], or iron [Fe] [Vit TE-Fe-]), solution 1 (Vit+TE+Fe-), solution 2 (Vit+TE-Fe-), solution 3 (Vit-TE+Fe ), solution 4 (Vit-TE-Fe+), and solution 5 (Vit+TE+Fe+). RESULTS: MDA concentrations in PN solutions were significantly higher at H24 than at H0 when they contained multivitamins (P < .001), trace elements (P = .002), or iron saccharate (P = .018). MDA concentration was particularly high when all 3 micronutrients were present (P < .001) or when the solutions were exposed to ambient light. In solutions containing iron, MDA concentrations were elevated at H0, and levels did not change whether protected from (P < .001) or exposed to (P < .001) from light. CONCLUSIONS: The addition of vitamins and trace elements to PN solutions induces a significant increase in peroxidation products, which are lowered when admixtures are protected from light. Iron should not be included in these solutions, even if solutions are light-protected. By following these conditions it is possible to use all-in-one admixtures in the nutrition management of preterm infants. PMID- 21632955 TI - Butyric acid: putting some fiber into your parenteral nutrition solution. PMID- 21632956 TI - Role for pAKT in rat urinary bladder with cyclophosphamide (CYP)-induced cystitis. AB - AKT phosphorylation following peripheral nerve injury or inflammation may play a role in somatic pain processes and visceral inflammation. To examine such a role in micturition reflexes with bladder inflammation, we induced bladder inflammation in adult female Wistar rats (200-300 g) by injecting cyclophosphamide (CYP) intraperitoneally at acute (150 mg/kg; 4 h), intermediate (150 mg/kg; 48 h), and chronic (75 mg/kg; every third day for 10 days) time points. Western blot analyses of whole urinary bladders showed significant increases (P <= 0.01) in phosphorylated (p) AKT at all time points; however, the magnitude of AKT phosphorylation varied with duration of CYP treatment. Immunohistochemical analyses of pAKT immunoreactivity (pAKT-IR) in cryostat bladder sections demonstrated duration-dependent, significant (P <= 0.01) increases in pAKT-IR in both the urothelium and detrusor smooth muscle of CYP inflamed bladders. Additionally, a suburothelial population of pAKT-IR macrophages (CD68-, MAC2-, and F4/80-positive) was present in chronic CYP-treated bladders. The functional role of pAKT in micturition was evaluated using open, conscious cystometry with continuous instillation of saline in conjunction with administration of an inhibitor of AKT phosphorylation, deguelin (1.0 MUg/10 MUl), or vehicle (1% DMSO in saline) in control (no inflammation) and CYP (48 h) treated rats. Bladder capacity, void volume, and intercontraction void interval increased significantly (P <= 0.05) following intravesical instillation of deguelin in CYP (48 h)-treated rats. These results demonstrate increased AKT phosphorylation in the urinary bladder with urinary bladder inflammation and that blockade of AKT phosphorylation in the urothelium improves overall bladder function. PMID- 21632958 TI - A new mouse model for Bartter's syndrome. PMID- 21632957 TI - Renovascular hypertension using a modified two-kidney, one-clip approach in mice is not dependent on the alpha1 or alpha2 Na-K-ATPase ouabain-binding site. AB - Endogenous cardiotonic steroids, through their interaction with the ouabain binding site of the Na-K-ATPase alpha-subunit, have been implicated in a variety of cardiovascular disease states including hypertension. We have previously shown that ACTH-induced hypertension is abolished in mutant mice expressing ouabain resistant alpha1- and alpha2-subunits. To further evaluate hypertension resistance in these mutant mice, we examined blood pressure changes in a modified model of 2-kidney, 1-clip (2K1C) renovascular hypertension. To reliably generate 2K1C hypertension, we used polyvinyl tubing (inner diameter: ~0.27 mm) to accurately gauge the degree of renal artery stenosis. Using this method, virtually all of the clipped mice became hypertensive and there was no incidence of apparent renal ischemia. By telemetry, in response to renal artery clipping, blood pressure in wild-type mice (alpha1 ouabain-resistant, alpha2 ouabain sensitive) increased from 97 +/- 3 to 136 +/- 7 mmHg. In alpha1-resistant, alpha2 resistant mice, pressure increased from 93 +/- 2 to 123 +/- 4 mmHg, and in alpha1 sensitive, alpha2-resistant mice, blood pressure increased from 95 +/- 2 to 139 +/- 5 mmHg. Blood pressure changes were equivalent in all three groups. In sham mice, blood pressure did not change (96 +/- 1 to 95 +/- 2 mmHg). Renin mRNA expression was dramatically elevated in the left vs. the right kidney, and plasma renin concentration was elevated similarly in all genotypes. These data indicate that sensitivity of the alpha1- or alpha2-Na-K-ATPase binding site to cardiotonic steroids is not a prerequisite for the development of 2K1C renovascular hypertension. In addition, use of a polyurethane cuff to constrict the renal artery provides a reliable method for producing 2K1C hypertension in mice. PMID- 21632960 TI - Two to tango: effects of collaboration and disagreement on dyadic judgment. AB - Four studies examined dyadic collaboration on quantitative estimation tasks. In accord with the tenets of "naive realism," dyad members failed to give due weight to a partner's estimates, especially those greatly divergent from their own. The requirement to reach joint estimates through discussion increased accuracy more than reaching agreement through a mere exchange of numerical "bids." However, even the latter procedure increased accuracy, relative to that of individual estimates (Study 1). Accuracy feedback neither increased weight given to partner's subsequent estimates nor produced improved accuracy (Study 2). Long term dance partners, who shared a positive estimation bias, failed to improve accuracy when estimating their performance scores (Study 3). Having dyad members ask questions about the bases of partner's estimates produced greater yielding and accuracy increases than having them explain their own estimates (Study 4). The latter two studies provided additional direct and indirect evidence for the role of naive realism. PMID- 21632961 TI - Mood and multiple source characteristics: mood congruency of source consensus status and source trustworthiness as determinants of message scrutiny. AB - This research deals with the interplay of mood and multiple source characteristics in regard to persuasion processes and attitudes. In a four factorial experiment, mood (positive vs. negative), source consensus status (majority vs. minority), source trustworthiness (high vs. low), and message strength (strong vs. weak) were manipulated. Results were in line with predictions of a mood-congruent expectancies perspective rather than competing predictions of a mood-as-information perspective. Specifically, individuals in both moods evinced higher message scrutiny given mood-incongruent (vs. mood congruent) source characteristics. That is, across source trustworthiness, positive (negative) mood led to higher message scrutiny given a minority (majority) versus a majority (minority) source. Furthermore, across source consensus, positive (negative) mood led to higher message scrutiny given an untrustworthy (trustworthy) versus a trustworthy (untrustworthy) source. Additional analyses revealed that processing effort increased from doubly mood congruent source combinations (low effort) over mixed-source combinations (intermediate effort) to doubly mood-incongruent combinations (high effort). Implications are discussed. PMID- 21632959 TI - Marker expression, behaviors, and responses vary in different lines of conditionally immortalized cultured podocytes. AB - The state-of-the-art cultured podocyte is conditionally immortalized by expression of a temperature-sensitive mutant of the SV40 large-T antigen. These cultures proliferate at 33 degrees C and differentiate at 37 degrees C into arborized cells that more closely resemble in vivo podocytes. However, the degree of resemblance remains controversial. In this study, several parameters were measured in podocyte cell lines derived from mouse (JR, KE), human (MS), and rat (HK). In all lines, the quantities of NEPH1 and podocin proteins and NEPH1 and SYNPO mRNAs were comparable to glomeruli, while synaptopodin and nephrin proteins and NPHS1 and NPHS2 mRNAs were <5% of glomerular levels. Expression of Wilms' tumor-1 (WT1) mRNA in mouse lines was comparable to glomeruli, but rat and human lines expressed little WT1. Undifferentiated human and mouse lines had similar proliferation rates that decreased after differentiation, while the rate in rat cells remained constant. The motility of different lines varied as measured by both general motility and wound-healing assays. The toxicity of puromycin aminonucleoside was MS ~ JR >> KE, and of doxorubicin was JR ~ KE > MS, while HK cells were almost unaffected. Process formation was largely a result of contractile action after formation of lamellipodia. These findings demonstrate dramatic differences in marker expression, response to toxins, and motility between lines of podocytes from different species and even between similarly derived mouse lines. PMID- 21632962 TI - Metabolism of methylisoeugenol in liver microsomes of human, rat, and bovine origin. AB - Methylisoeugenol (1,2-dimethoxy-4-propenylbenzene, 1) is a minor constituent of essential oils, naturally occurring as a mixture of cis/trans isomers. 1 is a U.S. Food and Drug Administration-approved food additive and has been given "Generally Recognized as Safe" status. Previously, metabolism of 1 has been studied in the rat, revealing mainly nontoxic cinnamoyl derivatives as major metabolites. However, data concerning the possible formation of reactive intermediary metabolites are not available to date. In this study, the oxidative metabolism of 1 was studied using liver microsomes of rat [not induced, rat liver microsomes (RLM); Aroclor1254 induced RLM (ARLM)], bovine, and human (pooled from 150 donors) origin. Incubations of these microsomes with 1 provided phase I metabolites that were separated by high-performance liquid chromatography (HPLC) and identified by NMR and UV-visible spectroscopy and/or liquid chromatography mass spectrometry. Identity was confirmed by comparison with (1)H NMR spectra of synthesized reference compounds. Formation of metabolites was quantified by HPLC/UV using dihydromethyleugenol (10) synthesized as the internal standard. From incubations of ARLM with 1, seven metabolites could be detected, with 3' hydroxymethylisoeugenol (2), isoeugenol and isochavibetol (3 + 4), and 6 hydroxymethylisoeugenol (5) being the main metabolites. Secondary metabolites derived from 1 were identified as the alpha,beta-unsaturated aldehyde 3' oxomethylisoeugenol (6) and 1',2'-dihydroxy-dihydromethylisoeugenol (7). We were surprised to find that formation of allylic 6-hydroxymethyleugenol (8) was observed starting at approximately 30 min after the beginning of incubations with ARLM. HLM did not form ring-hydroxylated metabolites but were most active in the formation of 6 and 7. ARLM incubations displayed the highest turnover rate and broadest metabolic pattern, presumably resulting from an increased expression of cytochrome P450 enzymes. In conclusion, we present a virtually complete pattern of nonconjugated microsomal metabolites of 1 comprising reactive metabolites and suggest the formation of reactive intermediates that need more investigation with respect to their possible adverse properties. PMID- 21632963 TI - Inhibitory effects of commonly used herbal extracts on UDP glucuronosyltransferase 1A4, 1A6, and 1A9 enzyme activities. AB - The aim of this study was to investigate the effect of commonly used botanicals on UDP-glucuronosyltransferase (UGT) 1A4, UGT1A6, and UGT1A9 activities in human liver microsomes. The extracts screened were black cohosh, cranberry, echinacea, garlic, ginkgo, ginseng, milk thistle, saw palmetto, and valerian in addition to the green tea catechin epigallocatechin gallate (EGCG). Formation of trifluoperazine glucuronide, serotonin glucuronide, and mycophenolic acid phenolic glucuronide was used as an index reaction for UGT1A4, UGT1A6, and UGT1A9 activities, respectively, in human liver microsomes. Inhibition potency was expressed as the concentration of the inhibitor at 50% activity (IC(50)) and the volume in which the dose could be diluted to generate an IC(50)-equivalent concentration [volume/dose index (VDI)]. Potential inhibitors were EGCG for UGT1A4, milk thistle for both UGT1A6 and UGT1A9, saw palmetto for UGT1A6, and cranberry for UGT1A9. EGCG inhibited UGT1A4 with an IC(50) value of (mean +/- S.E.) 33.8 +/- 3.1 MUg/ml. Milk thistle inhibited both UGT1A6 and UGT1A9 with IC(50) values of 59.5 +/- 3.6 and 33.6 +/- 3.1 MUg/ml, respectively. Saw palmetto and cranberry weakly inhibited UGT1A6 and UGT1A9, respectively, with IC(50) values >100 MUg/ml. For each inhibition, VDI was calculated to determine the potential of achieving IC(50)-equivalent concentrations in vivo. VDI values for inhibitors indicate a potential for inhibition of first-pass glucuronidation of UGT1A4, UGT1A6, and UGT1A9 substrates. These results highlight the possibility of herb-drug interactions through modulation of UGT enzyme activities. Further clinical studies are warranted to investigate the in vivo extent of the observed interactions. PMID- 21632964 TI - Gestation time-dependent pharmacokinetics of intravenous (+)-methamphetamine in rats. AB - We tested the hypothesis that differences in (+)-methamphetamine (METH) disposition during late rat pregnancy could lead to increased vulnerability to acute METH effects. The disposition of a single 1 mg/kg i.v. METH dose was studied during early (gestation day 7, GD7) and late (GD21) gestation. Results showed gestation time-dependent pharmacokinetics, characterized by a significantly higher area under the METH serum concentration versus time curve and a lower clearance on GD21 (p < 0.05; total, renal, and nonrenal clearance). The terminal elimination half-life (t(1/2lambdaz)) of METH and (+)-amphetamine (AMP; a pharmacologically active metabolite of METH) were not different on GD7, but by GD21, AMP t(1/2lambdaz) was 37% longer than METH t(1/2lambdaz) (p < 0.05). To identify the mechanism for AMP metabolite changes, intravenous AMP pharmacokinetics on GD21 were compared with AMP metabolite pharmacokinetics after intravenous METH. The intravenous AMP t(1/2lambdaz) was significantly shorter than metabolite AMP t(1/2lambdaz) (p < 0.05), which suggested AMP metabolite formation (not elimination) was the rate-limiting process. To understand the medical consequence of METH use during late-stage pregnancy, timed-pregnant rats received an intravenous dose of saline or METH (1, 3, or 5.6 mg/kg) on GD21, 0 to 2 days antepartum. Although one rat died and another had stillbirths at term after the 5.6-mg/kg dose, the pharmacokinetic values for all of the other animals were not significantly different. In conclusion, late-gestational clearance reductions lengthen METH exposure time, possibly increasing susceptibility to adverse effects, including death. PMID- 21632965 TI - Physiologically based pharmacokinetic modeling of intestinal first-pass metabolism of CYP3A substrates with high intestinal extraction. AB - Prediction of intestinal availability (F(G)), in conjunction with hepatic metabolism, is of considerable importance in drug disposition to assess oral clearance and liability to drug-drug interactions. In the current study, F(G) predictions were performed within a physiologically based pharmacokinetic (PBPK) model using in vitro permeability and clearance data. The prediction success was assessed in comparison with the Q(Gut) model. In addition, apparent oral clearance values, predicted using the PBPK model, were compared with in vivo observations from meta-analyses. Finally, unbound intrinsic clearance values (CLu(int)) were determined for 12 CYP3A substrates in eight individual human jejunal microsome (HJM) samples to assess interindividual variability in intestinal intrinsic clearance and subsequent F(G) predictions. Overall, the PBPK model improved F(G) predictions in comparison with the Q(Gut) model; this was apparent by a reduced bias and increased precision. In particular, F(G) predictions of indinavir, saquinavir, and terfenadine were model-dependent. The predicted oral clearance values of the drugs investigated ranged from 8.79 to 6320 l/h for tacrolimus and simvastatin, respectively, and were overall within 3 fold of the observed data with the exception of indinavir, atorvastatin, and buspirone. The individual HJM CLu(int) values ranged from 17 to 14,000 MUl . min( 1) . mg(-1) for atorvastatin and saquinavir, respectively, and corresponding interindividual variability in CLu(int) estimates ranged from 41 to 67%. These in vitro data resulted in predicted F(G) values ranging from 0.03 to 0.94 for simvastatin and indinavir, respectively. The largest interindividual variability of F(G) was predicted for terfenadine (65%) in contrast with the low variability in the case of indinavir (3%). PMID- 21632966 TI - "Out of my league": a real-world test of the matching hypothesis. AB - The matching hypothesis predicts that individuals on the dating market will assess their own self-worth and select partners whose social desirability approximately equals their own. It is often treated as well established, despite a dearth of empirical evidence to support it. In the current research, the authors sought to address conceptual and methodological inconsistencies in the extant literature and to examine whether matching occurs as defined by Walster et al. and more generally. Using data collected in the laboratory and from users of a popular online dating site, the authors found evidence for matching based on self-worth, physical attractiveness, and popularity, but to different degrees and not always at the same stage of the dating process. PMID- 21632967 TI - Explaining the paradoxical rejection-aggression link: the mediating effects of hostile intent attributions, anger, and decreases in state self-esteem on peer rejection-induced aggression in youth. AB - People are strongly motivated to feel accepted by others. Yet when faced with acute peer rejection they often aggress against the very peers they desire acceptance from, which may lead to further rejection. The present experiment tests three potential mediators of aggressive responses to acute peer rejection in the critical developmental stage of early adolescence. Participants (N=185, M(age)=11.5 years) completed personal profiles that were allegedly evaluated online by peers. After receiving negative or neutral peer feedback, participants could aggress against the same peers who had evaluated them. Rejected participants attributed more hostile intent to the peers, were angrier, showed a greater reduction in state self-esteem, and were more aggressive. Mediational analyses showed that hostile intent attributions mediated the acute peer rejection-aggression relationship, whereas increases in anger and decreases in state self-esteem did not. Thus, acute peer rejection evokes hostile intent attributions that, in turn, lead to aggressive reactions. PMID- 21632968 TI - The effects of self-criticism and self-oriented perfectionism on goal pursuit. AB - Five separate studies examined the associations of self-criticism and self oriented perfectionism with goal pursuit across a variety of domains. Although self-criticism has previously been shown to be related to diminished goal progress, a controversy remains regarding the potential association between aspects of "positive perfectionism," such as self-oriented perfectionism, and enhanced goal progress. The results of the five studies demonstrated a consistent pattern of negative association between self-criticism and goal progress. The results also showed a positive association between self-oriented perfectionism and goal progress when self-criticism was controlled. The important role of self criticism for understanding the impact of perfectionistic concerns is highlighted by these results. Implications for the debate concerning the possible positive effects of perfectionistic strivings are also discussed. PMID- 21632969 TI - Gynaecological symptoms reported by young women: examining the potential for earlier diagnosis of cervical cancer. AB - BACKGROUND: Cervical cancer occurs at a younger age than most adult cancers. A pre-malignant stage can be identified at screening and treated. Screening begins at the age of 25 years in England, so in women younger than this, and in those who decline screening, cervical cancer can only be identified with symptoms. Aim. To identify the frequency of attendance for gynaecological conditions by young English women. DESIGN: Historical cohort study using electronic primary care records. METHODS: A cohort of English women aged 15-29 years was prepared from the General Practice Research Database. All gynaecological consultations were identified and collated. Frequencies of gynaecological consultation were analysed in three age bands: 15-19, 20-24 and 25-29 years and by calendar year. RESULTS: The number of women available for study for each year ranged from 32 968 to 45 807. The percentage of women having any gynaecological consultation increased from 17.7% to 33.3% over the 7 years. If contraception is excluded, the percentages are 11.3% in 2003, rising to 20.1% in 2009. The rise in consultations occurred in all age bands and across most symptom categories. Post-coital bleeding and inter-menstrual bleeding-the two classic presentations of cervical cancer-were reported by 0.5% and 1.6% of women in 2009. CONCLUSIONS: Gynaecological complaints are frequent in primary care, though the symptoms of possible cervical cancer only represent a small minority of the total. Although the chance of cancer in young women with abnormal vaginal bleeding is very small, visualization of the cervix is appropriate. PMID- 21632970 TI - A shift toward smaller cell size via manipulation of cell cycle gene expression acts to smoothen Arabidopsis leaf shape. AB - Understanding the relationship of the size and shape of an organism to the size, shape, and number of its constituent cells is a basic problem in biology; however, numerous studies indicate that the relationship is complex and often nonintuitive. To investigate this problem, we used a system for the inducible expression of genes involved in the G1/S transition of the plant cell cycle and analyzed the outcome on leaf shape. By combining a careful developmental staging with a quantitative analysis of the temporal and spatial response of cell division pattern and leaf shape to these manipulations, we found that changes in cell division frequency occurred much later than the observed changes in leaf shape. These data indicate that altered cell division frequency cannot be causally involved in the observed change of shape. Rather, a shift to a smaller cell size as a result of the genetic manipulations performed correlated with the formation of a smoother leaf perimeter, i.e. appeared to be the primary cellular driver influencing form. These data are discussed in the context of the relationship of cell division, growth, and leaf size and shape. PMID- 21632971 TI - Tissue-specific differences in cytosine methylation and their association with differential gene expression in sorghum. AB - It has been well established that DNA cytosine methylation plays essential regulatory roles in imprinting gene expression in endosperm, and hence normal embryonic development, in the model plant Arabidopsis (Arabidopsis thaliana). Nonetheless, the developmental role of this epigenetic marker in cereal crops remains largely unexplored. Here, we report for sorghum (Sorghum bicolor) differences in relative cytosine methylation levels and patterns at 5'-CCGG sites in seven tissues (endosperm, embryo, leaf, root, young inflorescence, anther, and ovary), and characterize a set of tissue-specific differentially methylated regions (TDMRs). We found that the most enriched TDMRs in sorghum are specific for the endosperm and are generated concomitantly but imbalanced by decrease versus increase in cytosine methylation at multiple 5'-CCGG sites across the genome. This leads to more extensive demethylation in the endosperm than in other tissues, where TDMRs are mainly tissue nonspecific rather than specific to a particular tissue. Accordingly, relative to endosperm, the other six tissues showed grossly similar levels though distinct patterns of cytosine methylation, presumably as a result of a similar extent of concomitant decrease versus increase in cytosine methylation that occurred at variable genomic loci. All four tested TDMRs were validated by bisulfite genomic sequencing. Diverse sequences were found to underlie the TDMRs, including those encoding various known-function or predicted proteins, transposable elements, and those bearing homology to putative imprinted genes in maize (Zea mays). We further found that the expression pattern of at least some genic TDMRs was correlated with its tissue specific methylation state, implicating a developmental role of DNA methylation in regulating tissue-specific or -preferential gene expression in sorghum. PMID- 21632972 TI - Arabidopsis SUMO E3 ligase SIZ1 is involved in excess copper tolerance. AB - The reversible conjugation of the small ubiquitin-like modifier (SUMO) to protein substrates occurs as a posttranslational regulatory process in eukaryotic organisms. In Arabidopsis (Arabidopsis thaliana), several stress-responsive SUMO conjugations are mediated mainly by the SUMO E3 ligase SIZ1. In this study, we observed a phenotype of hypersensitivity to excess copper in the siz1-2 and siz1 3 mutants. Excess copper can stimulate the accumulation of SUMO1 conjugates in wild-type plants but not in the siz1 mutant. Copper accumulated to a higher level in the aerial parts of soil-grown plants in the siz1 mutant than in the wild type. A dramatic difference in copper distribution was also observed between siz1 and wild-type Arabidopsis treated with excess copper. As a result, the shoot-to root ratio of copper concentration in siz1 is nearly twice as high as that in the wild type. We have found that copper-induced Sumoylation is involved in the gene regulation of metal transporters YELLOW STRIPE-LIKE 1 (YSL1) and YSL3, as the siz1 mutant is unable to down-regulate the expression of YSL1 and YSL3 under excess copper stress. The hypersensitivity to excess copper and anomalous distribution of copper observed in the siz1 mutant are greatly diminished in the siz1ysl3 double mutant and slightly in the siz1ysl1 double mutant. These data suggest that SIZ1-mediated sumoylation is involved specifically in copper homeostasis and tolerance in planta. PMID- 21632974 TI - To thy proteins be true: RNA editing in plants. PMID- 21632973 TI - BBX32, an Arabidopsis B-Box protein, functions in light signaling by suppressing HY5-regulated gene expression and interacting with STH2/BBX21. AB - A B-box zinc finger protein, B-BOX32 (BBX32), was identified as playing a role in determining hypocotyl length during a large-scale functional genomics study in Arabidopsis (Arabidopsis thaliana). Further analysis revealed that seedlings overexpressing BBX32 display elongated hypocotyls in red, far-red, and blue light, along with reduced cotyledon expansion in red light. Through comparative analysis of mutant and overexpression line phenotypes, including global expression profiling and growth curve studies, we demonstrate that BBX32 acts antagonistically to ELONGATED HYPOCOTYL5 (HY5). We further show that BBX32 interacts with SALT TOLERANCE HOMOLOG2/BBX21, another B-box protein previously shown to interact with HY5. Based on these data, we propose that BBX32 functions downstream of multiple photoreceptors as a modulator of light responses. As such, BBX32 potentially has a native role in mediating gene repression to maintain dark adaptation. PMID- 21632975 TI - An updated view on transcription factor GATA3-mediated regulation of Th1 and Th2 cell differentiation. AB - CD4 T(h) are critical for orchestrating adaptive immune responses. The expression of the transcription factor GATA3 (GATA-binding protein 3) is up-regulated or down-regulated during T(h)2 or T(h)1 cell differentiation, respectively. Furthermore, GATA3 is responsible for induction of T(h)2 differentiation and represses T(h)1 differentiation. In this review, we present an updated view on the molecular mechanisms through which GATA3 regulates T(h)1/T(h)2 differentiation. During T(h)2 cell differentiation, GATA3 directly binds to the T(h)2 cytokine gene locus at several regions and regulates expression. On the other hand, GATA3 inhibits T(h)1 cell differentiation by preventing up-regulation of IL-12 receptor beta2 and STAT4 (signal transducer and activator of transcription 4) and neutralization of Runx3 (runt-related transcription factor 3) function through protein-protein interaction. GATA3 may also directly act on the Ifng gene. In summary, GATA3 serves as a transcriptional activator or repressor through direct action on transcriptional machinery and/or affecting chromatin remodeling at many critical loci encoding cytokines, cytokine receptors, signaling molecules as well as transcription factors that are involved in the regulation of T(h)1 and T(h)2 differentiation. PMID- 21632976 TI - Ventilation tube treatment: a systematic review of the literature. AB - OBJECTIVE: The aim of this review was to study the effectiveness of ventilation tube (VT) treatment in children with secretory otitis media (SOM), assessed by improved hearing, normalized language and quality of life (QoL), and recurrent acute otitis media (rAOM), assessed by number of episodes of AOM and QoL. Data Sources. Cochrane Library, PubMed, and Embase databases were searched for randomized and nonrandomized controlled trials and cohort studies in English, Scandinavian, German, and French languages between 1966 and April 2007. Additional literature was retrieved from reference lists in the articles. REVIEW METHODS: A total of 493 abstracts were evaluated independently by 2 members of the project group, 247 full-text versions were assessed for inclusion criteria and quality using structured evaluation forms, and 63 articles were included in the review. RESULTS AND CONCLUSIONS: This review shows that there is strong scientific evidence (grade 1) that VT treatment of SOM improves hearing for at least 9 months and that QoL is improved for up to 9 months (grade 2 scientific evidence). There was insufficient evidence to support an effect of VT treatment for rAOM. There was also insufficient evidence to determine whether the design or material of the VT or the procedure used for insertion had any influence on the effect; however, there was some evidence (grade 3) that aspiration of secretion at insertion does not prolong VT treatment. Further research is needed to address these issues. PMID- 21632977 TI - Treatment of Jones fracture nonunions and refractures in the elite athlete: outcomes of intramedullary screw fixation with bone grafting. AB - BACKGROUND: Fractures of the proximal fifth metatarsal metaphysis (ie, the Jones fracture) can be problematic in the elite athlete because of a high incidence of nonunion and refracture with nonoperative treatment. Although these fractures are not common, athletes can suffer refracture or nonunion of a Jones fracture despite operative stabilization. This is often attributable to hardware of insufficient strength, aggressive postoperative rehabilitation, or biologic insufficiency at the fracture site. PURPOSE: The authors review the results of revision intramedullary screw fixation with cancellous autologous bone grafting or bone-marrow aspirate combined with demineralized bone matrix after refracture or nonunion of Jones fractures in elite athletes. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: The authors retrospectively reviewed the clinical and radiographic outcomes and return to sport in 21 elite athletes undergoing treatment of Jones fracture refractures or nonunions. All patients underwent intramedullary screw fixation with autologous bone graft (12 patients), bone marrow aspirate (BMA) + demineralized bone matrix (DBM) (8 patients), or no bone graft (1 patient). RESULTS: All athletes were able to return to their previous level of athletic competition at an average of 12.3 weeks. All fractures showed clinical and radiographic evidence of compete cortical healing. Only 1 patient subsequently suffered a refracture. CONCLUSION: The authors recommend revision fixation with a large, solid screw (5.5 mm or larger) and autologous bone grafting for symptomatic refractures and nonunions of the proximal fifth metatarsal in elite athletes. Additional investigation is needed to determine whether BMA combined with DBM is an effective substitute for cancellous autograft. PMID- 21632978 TI - Platelet-rich plasma protects tenocytes from adverse side effects of dexamethasone and ciprofloxacin. AB - BACKGROUND: Ruptured tendons heal very slowly and complete recovery from injury is uncertain. Platelet-rich plasma (PRP), a rich source of growth factors, is currently being widely tested as a soft tissue healing agent and may accelerate tendon repair. The authors assessed the ability of PRP to prevent in vitro adverse effects of 2 drugs commonly linked to tendon rupture and tendinopathy, glucocorticoids and fluoroquinolone antibiotics. HYPOTHESIS: The pro-healing response induced by PRP protects human tenocytes against the cytotoxic effects of dexamethasone and ciprofloxacin. STUDY DESIGN: Controlled laboratory study. METHODS: Human primary hamstring tenocytes were exposed to different doses of ciprofloxacin and dexamethasone with and without PRP. AlamarBlue, beta galactosidase assay, and live/dead stain were used to measure, respectively, viability, senescence, and death in tenocyte culture. RESULTS: The viability of cells exposed to high doses of ciprofloxacin was significantly decreased compared with controls, with no induced senescence but increased cell death. Dexamethasone reduced viable cell number without inducing overt cell death, but the number of senescent cells increased considerably. After co-treatment with 10% PRP, viable cell number increased significantly in both conditions and the number of dead cells decreased in ciprofloxacin-treated cultures. Moreover, dexamethasone induced senescence was markedly reduced by co-treatment with 10% PRP. CONCLUSION: This study demonstrates that ciprofloxacin and dexamethasone have differing adverse effects on human tenocytes, with ciprofloxacin inducing cell death while dexamethasone primarily induces senescence. The authors showed that PRP can protect cultured human tenocytes against cell death or senescence induced by these drugs. CLINICAL RELEVANCE: Both ciprofloxacin and dexamethasone are highly effective in treatment of inflammatory and infectious conditions, therefore new strategies to minimize their adverse effects are of strong interest. These findings suggest the potential for local administration of PRP to enhance tendon healing in patients undergoing glucocorticoid or fluoroquinolone treatment. PMID- 21632979 TI - Is hip muscle weakness a predisposing factor for patellofemoral pain in female novice runners? A prospective study. AB - BACKGROUND: Hip muscle weakness has been proposed to contribute to patellofemoral malalignment and the development of the patellofemoral dysfunction syndrome (PFDS). However, from the retrospective studies that have addressed this issue, it is still unclear if hip muscle weakness is a cause or a consequence of PFDS. PURPOSE: This study was undertaken to investigate if hip muscle weakness is a predisposing factor for the development of PFDS. STUDY DESIGN: Cohort study (prognosis); Level of evidence, 2. METHODS: Before the start of a 10-week "start to run" program, the isometric strength of the hip flexor, extensor, abductor, adductor, and external and internal rotator muscles was measured in 77 healthy female novice runners. During the 10-week training period, patellofemoral pain was diagnosed and registered by an orthopaedic surgeon. RESULTS: Statistical analysis revealed that there was no significant difference in strength of any of the assessed hip muscle groups between the runners who did and did not develop PFDS. Logistic regression analysis did not identify a deviation in strength of any of the assessed hip muscle groups as a risk factor for PFDS. CONCLUSION: The findings of this study suggest that isometric hip muscle strength might not be a predisposing factor for the development of PFDS. PMID- 21632980 TI - Letter to the editor: "Validating the requirement for beat-to-beat coupling of the Ca2+ clock and M clock in pacemaker cell normal automaticity". PMID- 21632981 TI - Comparison of phenotypic and transcriptomic effects of false-positive genotoxins, true genotoxins and non-genotoxins using HepG2 cells. AB - The conventional in vitro assays for genotoxicity assessment of chemicals are characterised by a high false-positive rate, thus failing to correctly predict their in vivo genotoxic effects. This study aimed to identify the cellular mechanisms induced by the false-positive genotoxins quercetin, 8-Hydroxyquinoline and 17-beta oestradiol in comparison to true genotoxins and non-genotoxins, by combining in vitro phenotypic parameters with transcriptomics data from HepG2 cells. The effects of these compounds on the phosphorylation of H2AX, cell cycle distribution and whole genome gene expression following treatment for 12, 24 and 48 h were compared with the effects of true genotoxins [benzo[a]pyrene and aflatoxin B1] and non-genotoxins (2,3,7,8-tetrachlorodibenzodioxin, cyclosporin A and ampicillin C). Quercetin induced similar phenotypic effects as true genotoxins and to some extent similar gene expression alterations. Different gene expression changes were also observed, including the up-regulation of DNA repair related genes. 8-Hydroxyquinoline and 17-beta oestradiol showed no similarities to the true genotoxins at both the phenotypic and the transcriptomic level. In a classification approach, classifiers were selected to discriminate between genotoxins and non-genotoxins. Subsequent analysis for the false-positive compounds showed quercetin to be predicted as genotoxic and 8-hydroxyquinoline and 17-beta oestradiol as non-genotoxic. Our results support that transcriptomics analysis of compound effects in HepG2 leads to similar results with phenotypic analysis and provides additional mechanistic information. Therefore, combined evaluation of gene expression alterations and relevant functional end points using HepG2 cells may contribute to the better understanding of modes-of-action of chemicals and the correct evaluation of their genotoxic properties. PMID- 21632982 TI - Cuticular hydrocarbons rather than peptides are responsible for nestmate recognition in Polistes dominulus. AB - A colony of social insects is like a fortress where access is allowed only to colony members. The epicuticular mixture of hydrocarbons has been widely reported to be involved in nestmate recognition in insects. However, recent studies have shown that polar compounds (mainly peptides) are also present, mixed with hydrocarbons, on the cuticle of various insects, including the paper wasps of the genus Polistes. As these polar compounds are variable among Polistes species and are perceived by the wasps, this cuticular fraction could also be involved in nestmate recognition. Through MALDI-TOF (Matrix-Assisted Laser Desorption Ionization Time of Flight) mass spectrometry analysis, we assessed, for the first time, the intercolonial variability of the cuticular polar fraction of Polistes dominulus in order to evaluate its reliability as source of nestmate recognition cues. We then tested through behavioral assays the importance of the 2 isolated fractions (apolar and polar) in nestmate recognition by presenting them separately to colonies of P. dominulus. Our results showed that the cuticular polar compounds are not colony specific and they are not used by paper wasps to discriminate nestmates from non-colony members. On the contrary, we confirmed that the isolated cuticular hydrocarbons are the chemical mediators prompting nestmate recognition in paper wasps. PMID- 21632983 TI - Rituximab's new therapeutic target: the podocyte actin cytoskeleton. AB - Therapeutic off-target activities are well recognized for small-molecule drugs. In contrast, monoclonal antibodies (mAbs) traditionally are believed to act specifically and lack off-target therapeutic effects. In this issue of Science Translational Medicine, Fornoni et al. show therapeutic benefit, through an off target-mediated mechanism, of the mAb drug rituximab in recurrent focal segmental glomerulosclerosis (FSGS) after kidney transplantation. These data shed new light on FSGS pathogenesis and suggest new therapeutic interventions for proteinuric diseases. PMID- 21632984 TI - Rituximab targets podocytes in recurrent focal segmental glomerulosclerosis. AB - Focal segmental glomerulosclerosis (FSGS) is a glomerular disease characterized by proteinuria, progression to end-stage renal disease, and recurrence of proteinuria after kidney transplantation in about one-third of patients. It has been suggested that rituximab might treat recurrent FSGS through an unknown mechanism. Rituximab not only recognizes CD20 on B lymphocytes, but might also bind sphingomyelin phosphodiesterase acid-like 3b (SMPDL-3b) protein and regulate acid sphingomyelinase (ASMase) activity. We hypothesized that rituximab prevents recurrent FSGS and preserves podocyte SMPDL-3b expression. We studied 41 patients at high risk for recurrent FSGS, 27 of whom were treated with rituximab at time of kidney transplant. SMPDL-3b protein, ASMase activity, and cytoskeleton remodeling were studied in cultured normal human podocytes that had been exposed to patient sera with or without rituximab. Rituximab treatment was associated with lower incidence of posttransplant proteinuria and stabilization of glomerular filtration rate. The number of SMPDL-3b(+) podocytes in postreperfusion biopsies was reduced in patients who developed recurrent FSGS. Rituximab partially prevented SMPDL-3b and ASMase down-regulation that was observed in podocytes treated with the sera of patients with recurrent FSGS. Overexpression of SMPDL-3b or treatment with rituximab was able to prevent disruption of the actin cytoskeleton and podocyte apoptosis induced by patient sera. This effect was diminished in cultured podocytes where SMPDL-3b was silenced. Our study suggests that treatment of high-risk patients with rituximab at time of kidney transplant might prevent recurrent FSGS by modulating podocyte function in an SMPDL-3b-dependent manner. PMID- 21632985 TI - A HIF-regulated VHL-PTP1B-Src signaling axis identifies a therapeutic target in renal cell carcinoma. AB - Metastatic renal cell carcinoma (RCC) is a molecularly heterogeneous disease that is intrinsically resistant to chemotherapy and radiotherapy. Although therapies targeted to the molecules vascular endothelial growth factor and mammalian target of rapamycin have shown clinical effectiveness, their effects are variable and short-lived, underscoring the need for improved treatment strategies for RCC. Here, we used quantitative phosphoproteomics and immunohistochemical profiling of 346 RCC specimens and determined that Src kinase signaling is elevated in RCC cells that retain wild-type von Hippel-Lindau (VHL) protein expression. RCC cell lines and xenografts with wild-type VHL exhibited sensitivity to the Src inhibitor dasatinib, in contrast to cell lines that lacked the VHL protein, which were resistant. Forced expression of hypoxia-inducible factor (HIF) in RCC cells with wild-type VHL diminished Src signaling output by repressing transcription of the Src activator protein tyrosine phosphatase 1B (PTP1B), conferring resistance to dasatinib. Our results suggest that a HIF-regulated VHL-PTP1B-Src signaling pathway determines the sensitivity of RCC to Src inhibitors and that stratification of RCC patients with antibody-based profiling may identify patients likely to respond to Src inhibitors in RCC clinical trials. PMID- 21632987 TI - Ultrasound quality and efficiency: how to make your practice flourish. PMID- 21632986 TI - MF59 adjuvant enhances diversity and affinity of antibody-mediated immune response to pandemic influenza vaccines. AB - Oil-in-water adjuvants have been shown to improve immune responses against pandemic influenza vaccines as well as reduce the effective vaccine dose, increasing the number of doses available to meet global vaccine demand. Here, we use genome fragment phage display libraries and surface plasmon resonance to elucidate the effects of MF59 on the quantity, diversity, specificity, and affinity maturation of human antibody responses to the swine-origin H1N1 vaccine in different age groups. In adults and children, MF59 selectively enhanced antibody responses to the hemagglutinin 1 (HA1) globular head relative to the more conserved HA2 domain in terms of increased antibody titers as well as a more diverse antibody epitope repertoire. Antibody affinity, as inferred by greatly diminished (>=10-fold) off-rate constants, was significantly increased in toddlers and children who received the MF59-adjuvanted vaccine. Moreover, MF59 also improved antibody affinity maturation after each sequential vaccination against avian H5N1 in adults. For both pandemic influenza vaccines, there was a close correlation between serum antibody affinity and virus-neutralizing capacity. Thus, MF59 quantitatively and qualitatively enhances functional antibody responses to HA-based vaccines by improving both epitope breadth and binding affinity, demonstrating the added value of such adjuvants for influenza vaccines. PMID- 21632988 TI - Reliable measurement procedure of virtual touch tissue quantification with acoustic radiation force impulse imaging. AB - OBJECTIVES: The purpose of this study was to find a reliable procedure for performing virtual touch tissue quantification of the liver. METHODS: Twenty healthy volunteers were enrolled after informed consent was obtained. The liver and spleen of each volunteer were examined on the same day by two different sonologists. The shear wave velocity values at deep and superficial locations of the right and left lobes of the liver and the spleen were obtained before and after food consumption and with and without deep inspiration. The measurements were performed 5 times in each location, and the mean shear wave velocities were recorded. RESULTS: No statistically significant difference was found between the mean shear wave velocities obtained by the two operators. A difference was found between the velocities obtained in the deep portion of the right lobe (5.5 cm from the probe; mean +/- SD, 1.17 +/- 0.25 m/s on the intercostal scan and 1.35 +/- 0.45 m/s on the subcostal scan) and the superficial portion (3.5 cm; 1.56 +/- 0.43 and 1.74 +/- 0.74 m/s; P < .001; P < .002, respectively), and the values obtained in the deep portion of the right lobe on the intercostal scan were lower than those obtained on the subcostal scan (P < .05). There were no differences in the mean shear wave velocities of the liver before and after food consumption or with and without deep inspiration. The velocities of the spleen did not differ before and after food consumption (2.33 +/- 0.47 versus 2.39 +/- 0.58 m/s). CONCLUSIONS: In healthy people, mean shear wave velocities were dependent on the measurement site in the liver, and food consumption or deep inspiration did not affect velocity measurements of the liver or spleen. PMID- 21632989 TI - Nonshadowing echogenic foci in thyroid nodules: are certain appearances enough to avoid thyroid biopsy? AB - OBJECTIVES: The purpose of this study was to further classify nonshadowing echogenic foci and examine the association with malignancy. METHODS: This study received Institutional Review Board approval and was Health Insurance Portability and Accountability Act compliant. A total of 371 consecutive thyroid nodules were evaluated in 189 patients (177 female and 12 male; mean age, 59 years; range, 21 92 years). Eighty-six nodules (23%) measured 5 mm or larger and contained nonshadowing echogenic foci with a mean nodule diameter of 16 mm (5-66 mm). Blinded review of these nodules 12 months later was performed. Echogenic foci were classified as follows: showing a comet tail artifact (type 1), linear and brightly echogenic (type 2), round and indeterminate (type 3), and microcalcifications (type 4). All available thyroid sonograms and pathologic data were then reviewed. RESULTS: Nineteen nodules (22%) showed a classic comet tail artifact, with malignancy in 0 of 19. Six (32%) had negative pathologic results, and 9 (47%) had stable imaging follow-up (mean, 37 months). Twenty-nine nodules (34%) showed linear and brightly echogenic foci, with malignancy in 0 of 29. Fifteen (52%) had negative pathologic results, and 11 (38%) had stable imaging follow-up (mean, 34 months). Twenty-four nodules (28%) showed round and indeterminate echogenic foci, with 1 of 24 (4%) containing papillary carcinoma. Thirteen (54%) had negative pathologic results, and 8 (33%) had stable imaging follow-up (mean, 24 months). Fourteen nodules (16%) contained microcalcifications, with 4 of 14 (29%) containing papillary thyroid cancer. Nine (64%) had negative pathologic results, and 1 (7%) had stable imaging follow-up (63 months). CONCLUSIONS: Nonshadowing brightly echogenic linear foci with or without a comet tail artifact may be a benign finding. Confirmatory studies are needed for this result to be applied clinically. PMID- 21632990 TI - Correlation between osteopontin messenger RNA expression and microcalcification shown on sonography in papillary thyroid carcinoma. AB - OBJECTIVES: The aims of this study were to investigate the correlation between osteopontin expression and microcalcification seen on sonograms of human papillary thyroid carcinoma and to explore the mechanism of microcalcification in these tumors. METHODS: Real-time reverse transcription-polymerase chain reaction and immunohistochemistry were used to determine expression levels and locations of osteopontin in 56 pathologically verified papillary thyroid carcinoma nodules. Osteopontin expression correlated with microcalcification and other sonographic signs recorded preoperatively. RESULTS: Osteopontin expression in the papillary thyroid carcinoma group was significantly enhanced compared with the control group for both messenger RNA and protein levels (P < .05). Osteopontin expression was significantly related to microcalcification and lymph node metastasis (P < .05), as shown on sonography. CONCLUSIONS: Our results suggest a possible role for osteopontin in the formation of microcalcification in papillary thyroid carcinoma; the sonographic signs can reflect the pathologic and biological behavior of these tumors. PMID- 21632991 TI - Quantitative estimates of the variability of in vivo sonographic measurements of the mouse aorta for studies of abdominal aortic aneurysms and related arterial diseases. AB - OBJECTIVES: Burgeoning interest in reducing the morbidity and mortality associated with abdominal aortic aneurysms (AAAs) has led to experimental strategies to elucidate the disease process and attain pharmacologic regression using the apolipoprotein E(-/-) (ApoE(-/-)) mouse model of angiotensin-induced AAAs and in vivo sonography. However, the variability of in vivo sonographic measurements of the mouse aorta has not been established. Thus, our purpose was to determine quantitative estimates of the variability of in vivo sonographic measurements of the mouse aorta as a guide for the design and assessment of studies focused on regression of AAAs and related arterial diseases. METHODS: We used Bland-Altman, locally weighted scatterplot-smoothing regression, and resampling (bootstrapping) methods for variability analyses of multiple in vivo short- and long-axis sonographic measurements of ApoE(-/-) mouse aortas. We measured distinct aortic sites in vivo at the baseline and after angiotensin induced AAAs and ex vivo using digital calipers. RESULTS: We analyzed 236 data points from 10 male mice (14 weeks old; mean weight +/- SD, 29.7 +/- 1.6 g). Overall intramouse differences between short- and long-axis and in vivo and ex vivo measurements were 0.038 (95% confidence interval [CI], 0.031-0.046) and 0.085 (95% CI, 0.062-0.109) mm, respectively. Intermouse differences in short axis measurements were 0.047 (95% CI, 0.042-0.053), 0.049 (95% CI, 0.044-0.055), and 0.039 (95% CI, 0.036-0.042) mm for infrarenal, suprarenal, and thoracic measurements, respectively; differences in long-axis measurements were 0.054 (95% CI, 0.044-0.064), 0.029 (95% CI, 0.024-0.034), and 0.046 (95% CI, 0.037-0.054) mm. Bland-Altman and locally weighted scatterplot-smoothing analyses showed excellent agreement between measures with no variation in discrepancies vis-a-vis the target measurement. CONCLUSIONS: These data establish previously undefined estimates of measurement variability relevant for in vivo sonographic studies of AAA regression in a commonly studied mouse model. PMID- 21632992 TI - Evaluation of a new disinfection method for ultrasound probes used for regional anesthesia: ultraviolet C light. AB - OBJECTIVES: Ultrasound-guided regional anesthesia is commonly used for block placement. At present, the risk of cross contamination from probes is not well documented. To avoid transmission of infectious agents, several methods have been used for probe disinfection and protection. The aim of this study was to evaluate the antibacterial efficacy of a new high-level disinfection method based on ultraviolet C (UV-C) light under routine conditions after block placement with an unprotected probe. METHODS: The study was after approval by the local Ethics Committee. In the first part of the study, 15 ultrasound probes were exposed to a large inoculum of 3 bacteria. Ultraviolet C disinfection consisted of cleaning the probe with dry and disinfectant-impregnated paper followed by a 90-second UV C disinfection cycle in a decontamination chamber. A protocol was established to retrieve the probe with sterile gloves after opening the door of the chamber. In the second part, 50 blocks were placed with ultrasound-guided regional anesthesia. The skin was first prepared with an antiseptic solution, and sterile gel was applied; no covers were used to protect the probes. The blocks were then disinfected with UV-C light. Bacteriologic samples were collected before and after the UV-C method and inoculated on chocolate agar plates. RESULTS: During the first part of the study, all probes were infected after inoculation (>150 colony-forming units) but were considered sterile (<10 colony-forming units) after disinfection. During the second part of the study, all probes were considered sterile before and after disinfection. CONCLUSIONS: Ultraviolet C disinfection seems relevant for ultrasound-guided regional anesthesia just before block placement. It offers simple, fast, and effective high-level disinfection. Moreover, this method should obviate the use of sterile probe covers, which can improve echogenicity. PMID- 21632993 TI - Detection of subcutaneous and intramuscular air with sonography: a sensitive and specific modality. AB - OBJECTIVES: Soft tissue air may raise suspicion for several life-threatening illnesses. Physical examination has limited sensitivity in detecting air, and computed tomography and magnetic resonance imaging are time-consuming and expensive. Sonography can show soft tissue air, but the sensitivity and specificity in this setting are currently unknown. Therefore, the purpose of this study was to assess the performance characteristics of sonography in depicting the presence, amount, and affected tissue plane in a cadaver model of soft tissue air. METHODS: We conducted a single-blinded observational cadaver study. Cutaneous sites were selected and marked (~4 or 5 sites on each of 6 cadavers). An examiner injected air at some sites, recording both the depth (plane) and volume. A second examiner, who was blinded to the injection procedure, examined each site with sonography and recorded the presence or absence of air and the plane (subcutaneous or intramuscular). The results were compared, and statistical analysis was performed. RESULTS: Twenty-eight sites on 6 cadavers were used. Sites were not used if they were damaged or were within 10 cm of previous dissection. Air was injected in 20 of 28 sites; the remaining 8 were sham injections. Sonography showed air in all of the 20 sites with air injected. No air was detected in 7 of the 8 remaining sham sites, yielding 1 false-positive case. The plane was accurately assessed in all cases. Sensitivity for detection was 100%, and specificity was 87.5%. CONCLUSIONS: Sonography accurately showed the presence, amount, and anatomic plane of soft tissue air in cadavers with sensitivity of 100%. It warrants investigation as a screening test in patients at high risk for subcutaneous air from conditions such as necrotizing fasciitis and gas gangrene. PMID- 21632994 TI - Sonographic evaluation of femoral trochlear cartilage in patients with knee pain. AB - OBJECTIVES: The purpose of this study was to investigate whether routine clinical sonographic evaluation of femoral trochlear cartilage can identify abnormalities in patients with knee pain. METHODS: Short-axis sonograms of the femoral trochlear cartilage were obtained from 20 symptomatic knees in 20 consecutive patients (16 women and 4 men; mean age, 54 years; range, 35-75 years) and 20 knees in 10 asymptomatic control participants (7 women and 3 men; mean age, 52 years; range, 31-74 years). Articular cartilage thickness was measured at 3 locations in each knee, and thickness at each site was compared between patients and controls. Subsequently, images from patients and controls were randomly assorted and evaluated by 3 blinded musculoskeletal radiologists, who independently evaluated the femoral trochlear cartilage in each sonogram for cartilage clarity, grade, and presence or absence of cartilage calcifications, osteophytes, and subchondral bony irregularity. The radiologists were instructed to rate the most severe lesion in each evaluated region (medial trochlea, trochlear notch, and lateral trochlea). Cartilage clarity was defined as how well the cartilage borders could be distinguished from the overlying intra-articular soft tissues, and grade was defined as the severity of focal cartilaginous lesions. RESULTS: Using the Student t test, there was no significant difference (P > .05) between cartilage thickness in patients and controls for any location measured. However, using repeated measures analysis of variance, there were significant differences (P = .02) for both decreased cartilage clarity (df = 1/28; F = 5.76) and increased grade (df = 1/28; F = 5.77) in patients. There was also a nonsignificant (P > .05) trend toward more frequent calcifications, osteophytes, and bony irregularity in patients. CONCLUSIONS: Routine clinical sonography can identify femoral trochlear cartilage abnormalities in patients with knee pain and therefore can be a useful adjunct to other imaging tests for identifying arthritic changes in the knee. PMID- 21632995 TI - Clinical experience with contrast infusion sonography as an Essure confirmation test. AB - OBJECTIVES: The purpose of this study was to assess the clinical experience, including the feasibility, safety, compliance, and efficacy, of contrast infusion sonography as an Essure (Conceptus Inc, Mountain View, CA) confirmation test. METHODS: A retrospective chart review and telephone survey were conducted at an academic multispecialty group. The study participants were women with Essure intervention who underwent contrast infusion sonography, transvaginal sonography, and hysterosalpingography as Essure confirmation tests. The main outcome measures included the feasibility, safety, compliance, and efficacy of contrast infusion sonography as a first-line Essure confirmation test. RESULTS: A total of 118 women had successful bilateral Essure placement. Of the 63 who consented to contrast infusion sonography, 53 (84.1%) had proper bilateral placement and tubal occlusion and were encouraged to rely on Essure. Four were suspected of having unilateral tubal patency; hysterosalpingography in 2 confirmed bilateral tubal occlusion, and 2 were noncompliant with second-line hysterosalpingography. Three patients suspected of having unsatisfactory or uncertain placement on initial transvaginal sonography were encouraged to undergo hysterosalpingography in lieu of contrast infusion sonography, which confirmed unsatisfactory placement in 2 and proper placement and occlusion in 1. Three contrast infusion sonographic procedures could not be completed because of technical issues; therefore, contrast infusion sonography was feasible in 95.2% of the patients (60 of 63). No notable adverse events occurred. Only 17 patients were noncompliant with any confirmation test, yielding an overall compliance rate of 86% (101 of 118). No pregnancies occurred during 669 woman-months of follow-up. The average reimbursement for contrast infusion sonography was US$251.78. CONCLUSIONS: Preliminary clinical data suggest that contrast infusion sonography is a feasible, safe, and accurate Essure confirmation test, which is well accepted by patients. PMID- 21632996 TI - Antenatally diagnosed renal duplex anomalies: sonographic features and long-term postnatal outcome. AB - OBJECTIVES: The purpose of this study was to assess the diagnostic accuracy and infant outcomes of antenatally detected renal duplex anomalies. METHODS: An observational longitudinal study of cases of renal duplex anomalies diagnosed on prenatal sonography was conducted. Information on the prenatal sonographic findings, antenatal course, and perinatal outcome was obtained by reviewing the sonography reports and delivery records. Postnatal follow-up, including neonatal and infant evaluations, was obtained from hospital records. RESULTS: During a 7 year period, a total of 24 cases of renal duplex anomalies were identified prenatally. Three cases were excluded from subsequent analysis because of termination of pregnancy after the diagnosis of a severe brain abnormality, loss to follow-up, and an inability to confirm the prenatal diagnosis on neonatal renal sonography. Among the 21 confirmed cases, the presence of two separate renal pelvises was the most common prenatal sonographic feature (n = 15 [71%]), followed by dilatation of a single moiety with a dilated ipsilateral ureter or ureterocele (n = 6 [29%]). In 43% of cases, the renal duplex anomalies had no adverse associations, but in 48%, they were complicated by a urinary tract infection, and 48% of the infants required a surgical procedure during early childhood. CONCLUSIONS: Renal duplex anomalies can be accurately diagnosed by prenatal sonography even when there is minimal dilatation of the renal pelvis. Antenatal diagnosis allows planning of postnatal care, which may prevent urinary tract infections and renal function impairment. The natural history of antenatally diagnosed duplex anomalies seems to be overall benign, with urinary tract infections and the need for surgery being the most common complications in affected infants. PMID- 21632997 TI - Influence of different rotation angles in assessment of lung volumes by 3 dimensional sonography in comparison to magnetic resonance imaging in healthy fetuses. AB - OBJECTIVES: Three-dimensional (3D) sonographic volumetry is established in gynecology and obstetrics. Assessment of the fetal lung volume by magnetic resonance imaging (MRI) in congenital diaphragmatic hernias has become a routine examination. In vitro studies have shown a good correlation between 3D sonographic measurements and MRI. The aim of this study was to compare the lung volumes of healthy fetuses assessed by 3D sonography to MRI measurements and to investigate the impact of different rotation angles. METHODS: A total of 126 fetuses between 20 and 40 weeks' gestation were measured by 3D sonography, and 27 of them were also assessed by MRI. The sonographic volumes were calculated by the rotational technique (virtual organ computer-aided analysis) with rotation angles of 6 degrees and 30 degrees . To evaluate the accuracy of 3D sonographic volumetry, percentage error and absolute percentage error values were calculated using MRI volumes as reference points. Formulas to calculate total, right, and left fetal lung volumes according to gestational age and biometric parameters were derived by stepwise regression analysis. RESULTS: Three-dimensional sonographic volumetry showed a high correlation compared to MRI (6 degrees angle, R(2) = 0.971; 30 degrees angle, R(2) = 0.917) with no systematic error for the 6 degrees angle. Moreover, using the 6 degrees rotation angle, the median absolute percentage error was significantly lower compared to the 30 degrees angle (P < .001). The new formulas to calculate total lung volume in healthy fetuses only included gestational age and no biometric parameters (R(2) = 0.853). CONCLUSIONS: Three-dimensional sonographic volumetry of lung volumes in healthy fetuses showed a good correlation with MRI. We recommend using an angle of 6 degrees because it assessed the lung volume more accurately. The specifically designed equations help estimate lung volumes in healthy fetuses. PMID- 21632998 TI - Sonographic examination of the fetal vermis: tricks for obtaining the narrow midline target with 3-dimensional volume contrast imaging in the C plane. AB - Prenatal imaging of the fetal cerebellar vermis is challenging even for experienced examiners. We found that by aiming the ultrasound beam through the mastoid fontanel and then rotating the data set in a multiplanar reconstruction and applying volume contrast imaging in the C plane, we were consistently able to obtain images of the vermis in the standard midsagittal plane. Images of the fetal vermis suitable for morphologic evaluation were obtained in 408 of 414 cases (98.5%) at gestational ages of 18 weeks to 31 weeks 6 days; the examination time was only minimally increased. PMID- 21632999 TI - Fetal megacystis: differential diagnosis. AB - The purpose of our retrospective observational series was to determine whether the sonographic characteristics of fetal megacystic bladders can be used to reliably establish the most likely diagnosis in fetuses with this condition. The sonographic records of pregnant patients referred to our institutions over a 10 year period who were found on initial 2-dimensional sonography to be carrying fetuses with megacystis were examined for evidence of a keyhole sign, bladder thickness, amniotic fluid index, and fetal sex. When available, 3-/4-dimensional sonography, Doppler angiography, tomographic ultrasound imaging, virtual organ computer-aided analysis, and automatic volume calculation were used as part of the detailed fetal anatomic survey. Twenty fetuses with megacystis were identified. Seventeen were male; 2 were female; and 1 had ambiguous genitalia. All male fetuses with megacystis originally had a diagnosis of prune belly syndrome. The diagnosis for 10 male fetuses with a keyhole sign was changed to megacystis secondary to posterior urethral valves. The fetus with ambiguous genitalia had prune belly syndrome. One of the female fetuses had a diagnosis of urethral atresia, and the diagnosis for the other female fetus was megacystis microcolon-intestinal hypoperistalsis syndrome. In conclusion, in fetuses with megacystic bladders, it is possible to distinguish between cases with prune belly syndrome, posterior urethral valves, urethral atresia, and megacystis-microcolon intestinal hypoperistalsis syndrome by a detailed anatomic survey using 2- and 3 /4-dimensioinal sonographic techniques. PMID- 21633000 TI - Prenatal magnetic resonance imaging (MRI) findings of a foregut duplication cyst of the tongue: value of real-time MRI evaluation of the fetal swallowing mechanism. AB - Foregut duplication cysts of the oral cavity or lingual choristomas have a potential risk of airway obstruction. Two cases are reported that were initially detected by screening sonography. Further imaging with both static and real-time cine magnetic resonance imaging confirmed the lingual origin, relationship of the mass to fluid-filled spaces within the oral cavity, motion of the mass with the tongue during fetal swallowing, and airway patency. The additional information provided by magnetic resonance imaging aided in planning delivery and obviated the need for an ex utero intrapartum treatment procedure because airway patency was confirmed in both cases. PMID- 21633001 TI - Tall cell variant of papillary thyroid carcinoma: sonographic and clinical findings. AB - We retrospectively reviewed the clinical and sonographic features of 8 patients with 10 tall cell variants of papillary thyroid carcinoma. The mean age of the patients was 57 years (range, 34-72 years). The tumor sizes varied. Of the 8 patients, 5 had symptomatic masses, and 3 had incidentalomas. Three patients had recurrences and 1 died of pulmonary metastasis within a mean time of 30 months. The tall cell variants often appeared as microlobulated markedly hypoechoic nodules with microcalcifications and extrathyroidal extension on sonography and were always associated with lymph node metastasis. The tall cell variant of papillary thyroid carcinoma should be included in the differential diagnosis of an aggressive thyroid tumor with symptoms and cervical nodal metastasis. PMID- 21633002 TI - Latissimus dorsi costal tear in an elite handball player. PMID- 21633003 TI - Sonographic characterization of mixed crystal arthropathy due to monosodium urate and calcium pyrophosphate dihydrate: the triple-contour sign. PMID- 21633004 TI - Lymphangioma and lymphangiectasia of the breast mimicking inflammatory breast cancer. PMID- 21633005 TI - Term singleton pregnancy after conservative management of a complicated triplet gestation including a heterotopic cornual monochorionic twin pair. PMID- 21633007 TI - Extragonadal tumor and testicular microlithiasis. PMID- 21633008 TI - Terrestrial influences on carbon burial at sea. PMID- 21633009 TI - Negative feedback in the bone morphogenetic protein 4 (BMP4) synexpression group governs its dynamic signaling range and canalizes development. AB - What makes embryogenesis a robust and canalized process is an important question in developmental biology. A bone morphogenetic protein (BMP) morphogen gradient plays a key role in embryonic development, and we are beginning to understand how the self-regulating properties of its signaling circuitry ensure robust embryonic patterning. An unexplored question is why the BMP signaling circuit is organized as a modular synexpression group, with a prevalence of feedback inhibitors. Here, we provide evidence from direct experimentation and mathematical modeling that the synexpressed feedback inhibitors BAMBI, SMAD6, and SMAD7 (i) expand the dynamic BMP signaling range essential for proper embryonic patterning and (ii) reduce interindividual phenotypic and molecular variability in Xenopus embryos. Thereby, negative feedback linearizes signaling responses and confers robust patterning, thus promoting canalized development. The presence of negative feedback inhibitors in other growth factor synexpression groups suggests that these properties may constitute a general principle. PMID- 21633010 TI - Comparative oncogenomics identifies breast tumors enriched in functional tumor initiating cells. AB - The claudin-low subtype is a recently identified rare molecular subtype of human breast cancer that expresses low levels of tight and adherens junction genes and shows high expression of epithelial-to-mesenchymal transition (EMT) genes. These tumors are enriched in gene expression signatures derived from human tumor initiating cells (TICs) and human mammary stem cells. Through cross-species analysis, we discovered mouse mammary tumors that have similar gene expression characteristics as human claudin-low tumors and were also enriched for the human TIC signature. Such claudin-low tumors were similarly rare but came from a number of distinct mouse models, including the p53 null transplant model. Here we present a molecular characterization of 50 p53 null mammary tumors compared with other mouse models and human breast tumor subtypes. Similar to human tumors, the murine p53 null tumors fell into multiple molecular subtypes, including two basal like, a luminal, a claudin-low, and a subtype unique to this model. The claudin low tumors also showed high gene expression of EMT inducers, low expression of the miR-200 family, and low to absent expression of both claudin 3 and E cadherin. These murine subtypes also contained distinct genomic DNA copy number changes, some of which are similarly altered in their cognate human subtype counterpart. Finally, limiting dilution transplantation revealed that p53 null claudin-low tumors are highly enriched for TICs compared with the more common adenocarcinomas arising in the same model, thus providing a unique preclinical mouse model to investigate the therapeutic response of TICs. PMID- 21633011 TI - Renal phenotype in mice lacking the Kir5.1 (Kcnj16) K+ channel subunit contrasts with that observed in SeSAME/EAST syndrome. AB - The heteromeric inwardly rectifying Kir4.1/Kir5.1 K(+) channel underlies the basolateral K(+) conductance in the distal nephron and is extremely sensitive to inhibition by intracellular pH. The functional importance of Kir4.1/Kir5.1 in renal ion transport has recently been highlighted by mutations in the human Kir4.1 gene (KCNJ10) that result in seizures, sensorineural deafness, ataxia, mental retardation, and electrolyte imbalance (SeSAME)/epilepsy, ataxia, sensorineural deafness, and renal tubulopathy (EAST) syndrome, a complex disorder that includes salt wasting and hypokalemic alkalosis. Here, we investigated the role of the Kir5.1 subunit in mice with a targeted disruption of the Kir5.1 gene (Kcnj16). The Kir5.1(-/-) mice displayed hypokalemic, hyperchloremic metabolic acidosis with hypercalciuria. The short-term responses to hydrochlorothiazide, an inhibitor of ion transport in the distal convoluted tubule (DCT), were also exaggerated, indicating excessive renal Na(+) absorption in this segment. Furthermore, chronic treatment with hydrochlorothiazide normalized urinary excretion of Na(+) and Ca(2+), and abolished acidosis in Kir5.1(-/-) mice. Finally, in contrast to WT mice, electrophysiological recording of K(+) channels in the DCT basolateral membrane of Kir5.1(-/-) mice revealed that, even though Kir5.1 is absent, there is an increased K(+) conductance caused by the decreased pH sensitivity of the remaining homomeric Kir4.1 channels. In conclusion, disruption of Kcnj16 induces a severe renal phenotype that, apart from hypokalemia, is the opposite of the phenotype seen in SeSAME/EAST syndrome. These results highlight the important role that Kir5.1 plays as a pH-sensitive regulator of salt transport in the DCT, and the implication of these results for the correct genetic diagnosis of renal tubulopathies is discussed. PMID- 21633012 TI - Concerted regulation of myofiber-specific gene expression and muscle performance by the transcriptional repressor Sox6. AB - In response to physiological stimuli, skeletal muscle alters its myofiber composition to significantly affect muscle performance and metabolism. This process requires concerted regulation of myofiber-specific isoforms of sarcomeric and calcium regulatory proteins that couple action potentials to the generation of contractile force. Here, we identify Sox6 as a fast myofiber-enriched repressor of slow muscle gene expression in vivo. Mice lacking Sox6 specifically in skeletal muscle have an increased number of slow myofibers, elevated mitochondrial activity, and exhibit down-regulation of the fast myofiber gene program, resulting in enhanced muscular endurance. In addition, microarray profiling of Sox6 knockout muscle revealed extensive muscle fiber-type remodeling, and identified numerous genes that display distinctive fiber-type enrichment. Sox6 directly represses the transcription of slow myofiber-enriched genes by binding to conserved cis-regulatory elements. These results identify Sox6 as a robust regulator of muscle contractile phenotype and metabolism, and elucidate a mechanism by which functionally related muscle fiber-type specific gene isoforms are collectively controlled. PMID- 21633013 TI - Cerebral vasculopathy is a common feature in Aicardi-Goutieres syndrome associated with SAMHD1 mutations. PMID- 21633014 TI - High-precision dating of colonization and settlement in East Polynesia. PMID- 21633015 TI - Tuberculosis-associated chronic kidney disease. AB - Extrapulmonary tuberculosis (TB) account for approximately 15-20% of TB cases in immunocompetent patients. The genitourinary system is the third most commonly affected site. We report the case of a 20-year-old man admitted with fever, chills, dry cough, right flank pain, and oliguria who developed renal function loss. The pyelogram evidenced silence of the right kidney, and the abdominal and pelvic magnetic resonance showed significant dilation of the right pyelocaliceal system and proximal ureter. Biopsies of renal cortex and retroperitoneal lymph nodes showed caseous granuloma consistent with TB. Treatment was started with rifampicin, isoniazid, pyrazinamide, and ethambutol, and the patient presented a favorable outcome but with non-dialytic chronic kidney disease. This case illustrates a case of chronic kidney disease secondary to TB in a young, otherwise healthy man. PMID- 21633016 TI - Emerging need for parasitology education: training to identify and diagnose parasitic infections. PMID- 21633017 TI - A cluster of cutaneous leishmaniasis associated with human smuggling. AB - Cutaneous leishmaniasis (CL) is rarely seen in the United States, and the social and geographic context of the infection can be a key to its diagnosis and management. Four Somali and one Ethiopian, in U.S. Border Patrol custody, came to the United States by the same human trafficking route: Djibouti to Dubai to Moscow to Havana to Quito; and then by ground by Columbia/Panama to the United States-Mexico border where they were detained. Although traveling at different times, all five patients simultaneously presented to our institution with chronic ulcerative skin lesions at different sites and stages of evolution. Culture of biopsy specimens grew Leishmania panamensis. Soon thereafter, three individuals from East Africa traveling the identical route presented with L. panamensis CL to physicians in Tacoma, WA. We document here the association of a human trafficking route and new world CL. Clinicians and public health officials should be aware of this emerging infectious disease risk. PMID- 21633018 TI - The aftermath of the Western Australian melioidosis outbreak. AB - Melioidosis became a notifiable disease in Western Australia (WA) 2 years after the West Kimberley melioidosis outbreak. Two cases of melioidosis caused by the outbreak genotype of Burkholderia pseudomallei (National Collection of Type Cultures [NCTC] 13177) occurred in 1998 and 1999 in persons who visited the outbreak location at the time. No other infections caused by the outbreak strain have been recorded in WA since that time, despite an average of four culture positive cases per year. Sporadic cases of melioidosis often follow tropical storms and cyclones during summer, and they have been detected outside the endemic area when cyclones travel far inland. In 2007, environmental isolates resembling NCTC 13177 were found 500 km east of the outbreak location after unusually severe weather. Recent whole-genome analysis places NCTC 13177 genetically close to other Australian isolates. Additional biogeographic and ecological studies are needed to establish the relative importance of environmental cofactors in disease pathogenesis. PMID- 21633019 TI - Repeat blood culture positive for B. pseudomallei indicates an increased risk of death from melioidosis. AB - Melioidosis, a bacterial infection caused by Burkholderia pseudomallei, is notoriously difficult to cure despite appropriate antimicrobial therapy and has a mortality rate of up to 40%. We demonstrate that a blood culture positive for B. pseudomallei taken at the end of the first and/or second week after hospitalization for melioidosis is a strong prognostic factor for death (adjusted odds ratio = 4.2, 95% confidence interval = 2.1-8.7, P < 0.001 and adjusted odds ratio = 2.6, 95% confidence interval = 1.1-6.0, P = 0.03, respectively). However, repeat cultures of respiratory secretions, urine, throat swabs, or pus/surface swabs provide no prognostic information. This finding highlights the need for follow-up blood cultures in patients with melioidosis. PMID- 21633020 TI - Climate variability and the outbreaks of cholera in Zanzibar, East Africa: a time series analysis. AB - Global cholera incidence is increasing, particularly in sub-Saharan Africa. We examined the impact of climate and ocean environmental variability on cholera outbreaks, and developed a forecasting model for outbreaks in Zanzibar. Routine cholera surveillance reports between 1997 and 2006 were correlated with remotely and locally sensed environmental data. A seasonal autoregressive integrated moving average (SARIMA) model determined the impact of climate and environmental variability on cholera. The SARIMA model shows temporal clustering of cholera. A 1 degrees C increase in temperature at 4 months lag resulted in a 2-fold increase of cholera cases, and an increase of 200 mm of rainfall at 2 months lag resulted in a 1.6-fold increase of cholera cases. Temperature and rainfall interaction yielded a significantly positive association (P < 0.04) with cholera at a 1-month lag. These results may be applied to forecast cholera outbreaks, and guide public health resources in controlling cholera in Zanzibar. PMID- 21633021 TI - The variability of childhood diarrhea in Karachi, Pakistan, 2002-2006. AB - Diarrhea burden is often estimated using cross-sectional surveys. We measured variability in diarrhea prevalence among children < 5 years of age living in squatter settlements in central Karachi, Pakistan. We pooled data from non intervention control households from studies conducted from 2002 through 2006. The prevalence of diarrhea varied on average by 29% from one week to the next, by 37% from one month to the next, and during peak diarrhea season by 32% from one year to the next. During 24 months when the same nine neighborhoods were under surveillance, each month the prevalence of diarrhea varied by at least an order of magnitude from the lowest to the highest prevalence neighborhood, and each neighborhood recorded the highest diarrhea prevalence during at least one month. Cross-sectional surveys are unreliable measures of diarrhea prevalence. PMID- 21633022 TI - AIDS diarrhea and antiretroviral drug concentrations: a matched-pair cohort study in Port au Prince, Haiti. AB - Diarrhea in patients with acquired immunodeficiency syndrome (AIDS) may cause malabsorption of medications and failure of antiretroviral therapy (ART). We prospectively evaluated human immunodeficiency virus-1 (HIV-1)-infected patients with and without chronic diarrhea initiating ART in Haiti. We report mean plasma antiretroviral concentrations at 2 and 4 weeks. We measured plasma HIV-1 RNA levels at four points. Fifty-two HIV-1-infected patients (26 matched pairs) were enrolled. No differences in antiretroviral concentrations were detected. At week 24, 18/25 (72%) cases and 16/24 (68%) controls had undetectable plasma HIV-1 RNA levels (P = 0.69). Patients with plasma HIV-1 RNA levels > 50 copies/mL at week 24 had lower early efavirenz concentrations than patients with undetectable HIV-1 RNA (2,621 ng/mL versus 5,278 ng/mL; P = 0.02). Diarrhea at ART initiation does not influence plasma concentrations of the medications evaluated. Virologic outcome at Week 24 does correlate with efavirenz concentrations early in therapy but not with the presence of chronic diarrhea. PMID- 21633023 TI - Blastocystis sp. subtype 4 is common in Danish Blastocystis-positive patients presenting with acute diarrhea. AB - Fecal samples from 444 Danish patients presenting with acute diarrhea were tested for Blastocystis and positive samples were subtyped to investigate the prevalence and subtype distribution of Blastocystis in this patient group. A total of 25 patients (5.6%) were positive, and 19 of these patients (76.0%) were positive for Blastocystis sp. ST4. Because the relative prevalence of ST4 in other patients presenting with other types of diarrhea (persistent, travel-related, and human immunodeficiency virus-related) in Denmark is low, the role of Blastocystis sp. ST4 in the etiology of acute diarrhea should be investigated further. PMID- 21633024 TI - Enteric pathogens associated with childhood diarrhea in Tripoli-Libya. AB - Stool samples from children < 5 years of age with diarrhea (N = 239) were examined for enteric pathogens using a combination of culture, enzyme immunoassay, and polymerase chain reaction methods. Pathogens were detected in 122 (51%) stool samples; single pathogens were detected in 37.2% and co-pathogens in 13.8% of samples. Norovirus, rotavirus, and diarrheagenic Escherichia coli (DEC) were the most frequently detected pathogens (15.5%, 13.4%, and 11.2%, respectively); Salmonella, adenovirus, and Aeromonas were detected less frequently (7.9%, 7.1%, and 4.2%). The most commonly detected DEC was enteroaggregative E. coli (5.4%). Resistance to >= 3 antimicrobials was observed in 60% (18/30) of the bacterial pathogens. Salmonella resistance to ciprofloxacin (63.1%) has become a concern. Enteric viral pathogens were the most significant causative agents of childhood diarrhea in Tripoli. Bacterial pathogens were also important contributors to pediatric diarrhea. The emergence of ciprofloxacin resistant Salmonella represents a serious health problem that must be addressed by Libyan health authorities. PMID- 21633025 TI - Pharmacokinetics of oral sitamaquine taken with or without food and safety and efficacy for treatment of visceral leishmaniais: a randomized study in Bihar, India. AB - This randomized, open-label study of patients in India with visceral leishmaniasis (VL) investigated the effect of food on sitamaquine and desethyl sitamaquine pharmacokinetics. Patients were randomized to receive oral sitamaquine, 2 mg/kg/day, once a day for 21 days across four cohorts (n = 41) (fasted/fed, fed/fasted, fed/fed, and fasted/fasted) over two periods (days 1-10 and 11-21), or intravenous amphotericin B (AmB), 1 mg/kg every other day for 30 days (n = 20). Mean day 21 pharmacokinetics across the four cohorts were sitamaquine, area under curve (AUC)((0-tau)) = 6,627-8,903 ng.hr/mL, AUC((0-16)) = 4,859-6,633 ng.hr/mL, maximum plasma concentration (C(max)) = 401-570 ng/mL, apparent terminal half-life (t(1/2)) = 18.3-22.8 hr, time to reach C(max) (t(max)) = 3.5-6 hr; and desethyl-sitamaquine, AUC((0-tau)) = 2,307-3,163 ng.hr/mL, C(max) = 109-154 ng/mL, t(1/2) = 23.0-27.9 hr, t(max) = 2-10 hr, with no significant food effect. On-therapy adverse events were observed for sitamaquine in 4 (10%) of 41 patients and for AmB in 17 (85%) of 20 patients. The final clinical cure (day 180) was 85% (95% confidence interval = 70.8-94.4%) for sitamaquine and 95% (95% confidence interval = 75.1-99.9) for AmB. Sitamaquine can be taken regardless of food intake, was generally well tolerated, and showed potential efficacy in patients with visceral leishmaniasis. PMID- 21633026 TI - Endemic tegumentary leishmaniasis in Brazil: correlation between level of endemicity and number of cases of mucosal disease. AB - The purpose of this study was to establish a correlation between the endemic level of tegumentary leishmaniasis in different regions of Brazil during 2002 2009 and the number of cases of mucosal or mucocutaneous leishmaniasis. The proportion of mucosal leishmaniasis was inversely correlated with prevalence of infection. In areas with a lower infection prevalence, the proportion of mucosal leishmaniasis increased (P < 0.05). The hypothesis of an Amazonian origin and dissemination through human migration is considered. Our results show that in regions with lower prevalence and endemically younger, the proportion of cases that evolve to the mucosal form is higher than in regions with higher prevalence and endemically older. PMID- 21633027 TI - Disseminated cutaneous leishmaniasis resembling post-kala-azar dermal leishmaniasis caused by Leishmania donovani in three patients co-infected with visceral leishmaniasis and human immunodeficiency virus/acquired immunodeficiency syndrome in Ethiopia. AB - We report paired strains of Leishmania parasites, one from the viscera and the other from skin lesions that were isolated from three patients with visceral leishmaniasis and disseminated cutaneous leishmaniasis that were co-infected with human immunodeficiency virus. The causative parasites were characterized by polymerase chain reaction-restriction length polymorphism of the ribosomal DNA internal transcribed spacer 1 and by a panel of multilocus microsatellite markers. We demonstrated that the causative agent was Leishmania donovani in all cases, irrespective of the phenotype of the disease. The paired strains from viscera and skin lesions of the same patients showed genetic identity across the 14 microsatellite markers investigated. These findings demonstrate that the skin lesions in these human immunodeficiency virus-positive patients with visceral leishmaniasis were caused by dissemination of viscerotropic L. donovani parasites as a consequence of severe immunosuppression. However, in all three patients, rapid clearance of the skin lesions was observed after antimonial therapy. PMID- 21633028 TI - Phylogenetics of the phlebotomine sand fly group Verrucarum (Diptera: Psychodidae: Lutzomyia). AB - Within the sand fly genus Lutzomyia, the Verrucarum species group contains several of the principal vectors of American cutaneous leishmaniasis and human bartonellosis in the Andean region of South America. The group encompasses 40 species for which the taxonomic status, phylogenetic relationships, and role of each species in disease transmission remain unresolved. Mitochondrial cytochrome c oxidase I (COI) phylogenetic analysis of a 667-bp fragment supported the morphological classification of the Verrucarum group into series. Genetic sequences from seven species were grouped in well-supported monophyletic lineages. Four species, however, clustered in two paraphyletic lineages that indicate conspecificity--the Lutzomyia longiflocosa-Lutzomyia sauroida pair and the Lutzomyia quasitownsendi-Lutzomyia torvida pair. COI sequences were also evaluated as a taxonomic tool based on interspecific genetic variability within the Verrucarum group and the intraspecific variability of one of its members, Lutzomyia verrucarum, across its known distribution. PMID- 21633029 TI - Decreased susceptibility to commonly used antimicrobial agents in bacterial pathogens isolated from urinary tract infections in Rwanda: need for new antimicrobial guidelines. AB - The aim of this study was to obtain data on susceptibility patterns of pathogens responsible for both community and hospital urinary tract infections (UTIs); and analyzed risk factors for infection caused by ciprofloxacin-resistant Escherichia coli and extended-spectrum beta-lactamase (ESBL)-producing strains in Rwanda. Of 1,012 urine cultures prospectively studied, a total of 196 (19.3%) yielded significant growth of a single organism. The most common isolate (60.7%) was Escherichia coli. The antibiotics commonly used in UTIs are less effective except Fosfomycin-trometamol and imipinem. The use of ciprofloxacin in the previous 6 months (odds ratio [OR] = 7.59 [1.75-32.74]), use of other antibiotics in the previous 6 months (OR = 1.02 [1.02-2.34]), and production of ESBL (OR = 19.32 [2.62-142.16]) were found to be associated with ciprofloxacin resistance among the E. coli isolates. Risk factors for ESBL positivity were the use of ciprofloxacin and third-generation cephalosporin in the preceding 6 months (OR = 3.05 [1.42-6.58] and OR = 9.78 [2.71-35.25], respectively); and being an inpatient (OR = 2.27 [1.79-2.89]). Fosfomycin-trometamol could be included as a reasonable alternative for the therapy of uncomplicated UTI in Rwanda. PMID- 21633030 TI - Diagnosis of gestational, congenital, and placental malaria in Colombia: comparison of the efficacy of microscopy, nested polymerase chain reaction, and histopathology. AB - The technical capability of different methods to diagnose Plasmodium in maternal peripheral blood, placenta, and umbilical cord blood has not been assessed in Colombia and seldom explored in other malaria-endemic regions. We designed a study to compare the technical and the operational-economical performances of light microscopy (LM), nested polymerase chain reaction (nPCR), and histopathology (HP). In maternal blood, LM had 41% sensitivity and 100% specificity and in placental blood, 35% and 100%, respectively, compared with nPCR. In placental tissue, LM had 33% sensitivity and 95% specificity; and nPCR 47% and 77%, respectively; compared with HP. Light microscopy had the best operational-economical qualification. We concluded that nPCR and HP performed better compared with LM, but field implementation of these two techniques remains a problem. Therefore, LM is recommended as the gold standard for diagnosis of gestational malaria and placental blood infection in the field. PMID- 21633031 TI - Therapeutic efficacies of artemisinin-based combination therapies in Nigerian children with uncomplicated falciparum malaria during five years of adoption as first-line treatments. AB - The therapeutic efficacies of 3-day regimens of artesunate-amodiaquine and artemether-lumefantrine during 5 years of adoption as first-line treatments were evaluated in 811 <= 12-year-old malarious children. Compared with artemether lumefantrine, amodiaquine-artesunate significantly reduced the proportion of children with fever and parasitemia 1 day after treatment (day 1; P < 0.008 for both). The proportion of parasitemic children on day 2 and gametocytemia on presentation and carriage reduced significantly over the years (P < 0.000001 and P < 0.03, respectively; test for trend). Overall efficacy was 96.5% (95% confidence interval [CI] = 94.5-98.6) and remained unchanged over the years (P = 0.87; test for trend). Kinetics of parasitemias after treatments were estimated by a non-compartmental model. Declines of parasitemias were monoexponential, with a mean elimination half-life of 1.09 hours (95% CI = 1.0-1.16). Parasitemia half lives and efficacy were similar for both regimens and in all ages. Artesunate amodiaquine and artemether-lumefantrine remain efficacious treatments of uncomplicated falciparum malaria in Nigerian children 5 years after adoption. PMID- 21633032 TI - Antibody responses and avidity of naturally acquired anti-Plasmodium vivax Duffy binding protein (PvDBP) antibodies in individuals from an area with unstable malaria transmission. AB - Plasmodium vivax remains an important cause of morbidity outside Africa, and no effective vaccine is available against this parasite. The P. vivax Duffy binding protein (PvDBP) is essential during merozoite invasion into erythrocytes, and it is a target for protective immunity against malaria. This investigation was designed to evaluate naturally acquired antibodies to two variant forms of PvDBP II antigen (DBP-I and -VI) in malaria individuals (N = 85; median = 22 years) who were living in hypoendemic areas in Iran. The two PvDBP-II variants were expressed in Escherichia coli, and immunoglobulin G (IgG) isotype composition and avidity of naturally acquired antibodies to these antigens were measured using enzyme-linked immunosorbent assay (ELISA). Results showed that almost 32% of the studied individuals had positive antibody responses to the two PvDBP-II variants, and the prevalence of responders did not differ significantly (P > 0.05; chi(2) test). The IgG-positive samples exhibited 37.03% and 40.8% high-avidity antibodies for PvDBP-I and PvDBP-VI variants, respectively. Furthermore, high avidity IgG1 antibody was found in 39.1% of positive sera for each examined variant antigen. The avidity of antibodies for both PvDBP variant antigens and the prevalence of responders with high- and intermediate-avidity IgG, IgG1, and IgG3 antibodies were similar in patients (P > 0.05; chi(2) test). Moreover, the prevalence of IgG antibody responses to the two variants significantly increased with exposure and host age. To sum up, the results provided additional data in our understanding of blood-stage immunity to PvDBP, supporting the rational development of an effective blood-stage vaccine based on this antigen. PMID- 21633033 TI - Net risk: a risk assessment of long-lasting insecticide bed nets used for malaria management. AB - Despite the demonstrated ability of bed nets that have been factory-impregnated with long-lasting insecticides (LLINs) to protect people from malaria and despite the ambitious plans for their widespread use, the health risks from the LLINs themselves have not been adequately investigated and reported in the peer reviewed science literature. Here, we use a probabilistic risk assessment approach to estimate the risks to Africans from inhalation, dermal, and oral exposures to the newer LLINs with permethrin, alpha-cypermethrin, or deltamethrin as the insecticide active ingredient. We estimated exposures to LLINs using 17 age groups to incorporate different body weights and sleeping behaviors. Risk quotients (exposure divided by toxic threshold) at the 50th and 90th percentiles for non-cancer risks were < 1.0 for lifetime adjusted risk and all youth and adult age groups. Risk quotients for infants and toddlers (0-3 years) and child groups from 3 to 10 years were >= 1.0 for specific bed nets. PMID- 21633034 TI - Dexamethasone effects in the Strongyloides venezuelensis infection in a murine model. AB - The aim of this study was to investigate the immunomodulatory effects of glucocorticoids on the immune response to Strongyloides venezuelensis in mice. Balb/c mice were infected with S. venezuelensis and treated with Dexamethasone (Dexa) or vehicle. Dexa treatment increased circulating blood neutrophil numbers and inhibited eosinophil and mononuclear cell accumulation in the blood, bronchoalveolar, and peritoneal fluid compared with control animals. Moreover, Dexa decreased tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN gamma), interleukin-3 (IL-3), IL-4, IL-5, IL-10, and IL-12 production in the lungs and circulating immunoglobulin G1 (IgG1), IgG2a, and IgE antibody levels while increasing the overall parasite burden in the feces and intestine. Dexa treatment enhanced the fertility of female nematodes relative to untreated and infected mice. In summary, the alterations in the immune response induced by Dexa resulted in a blunted, aberrant immune response associated with increased parasite burden. This phenomenon is similar to that observed in S. stercoralis infected humans who are taking immunosuppressive or antiinflammatory drugs, including corticosteroids. PMID- 21633035 TI - Analysis of the cross-reactivity of various 56 kDa recombinant protein antigens with serum samples collected after Orientia tsutsugamushi infection by ELISA. AB - Orientia tsutsugamushi, the etiologic agent of scrub typhus, has a highly expressed and immunodominant 56-kD outer membrane protein. This protein is one of the leading candidates for diagnosis and vaccine development for scrub typhus. Previous studies using recombinant 56-kD protein (r56s) derived from Karp strain (Kpr56) in a mouse model have shown good homologous protection but only moderate to poor heterologous protection. We evaluated the cross-reactivity of recombinant 56-kD proteins from Karp, Kato, Gilliam, TA763, and three chimeric 56-kD proteins. Not all r56s are equally reactive with strain-specific serum samples. These data provide a first glance of how reactive these r56s are toward the antiserum of different strains and which r56 exhibits the broadest reactivity. A formulation of this combination has the potential to provide broad protection against the heterologous challenge and to be used in a highly sensitive diagnostic assay. PMID- 21633036 TI - A patient with erythema nodosus leprosum and Chagas cardiopathy: challenges in patient management and review of the literature. AB - We report a patient with severe multi-bacillary leprosy complicated by recurrent episodes of erythema nodosum necrotisans that required thalidomide and/or corticosteroids during follow-up. Although the patient was from an area to which Chagas disease is endemic, this diagnosis was initially missed and was only investigated when heart failure developed in the patient. The difficulties of managing erythema nodosum necrotisans and heart failure concomitantly and those involved in excluding the diagnosis of acute myocarditis caused by reactivation of Chagas disease secondary to the immunosuppressive regimen are discussed. Other potential causes for the heart failure and possible interactions between the two diseases and their treatments are discussed. We also reviewed the literature for the association between leprosy and Chagas disease, both of which are highly endemic in Brazil. This case emphasizes the importance of searching for subclinical co-infections in leprosy patients because reactions frequently develop during specific treatment in these patients, and these reactions require prolonged therapy with immunosuppressive drugs. PMID- 21633037 TI - Efficacy of a tetravalent chimeric dengue vaccine (DENVax) in Cynomolgus macaques. AB - Three tetravalent formulations of chimeric dengue (DENVax) viruses containing the pre-membrane and envelope genes of serotypes 1-4 expressed by the attenuated DENV 2 PDK-53 genome were tested for safety, immunogenicity, and efficacy in cynomolgus macaques (Macaca fascicularis). Subcutaneous injection of the DENVax formulations was well-tolerated. Low levels of viremia of only one of the four vaccine viruses were detected yet virus neutralizing antibody titers were induced against all four dengue virus serotypes after one or two administrations of vaccine. All animals immunized with the high-dose formulation were protected from viremia, and all immunized animals were completely protected from DENV-3 and DENV 4 challenge. A lower dose of DENVax formulation partially protected animals from DENV-1 or DENV-2 challenge. In contrast, all control animals developed high levels of viremia for multiple days after challenge with DENV 1-4. This study highlights the immunogenicity and efficacy of the tetravalent DENVax formulations in nonhuman primates. PMID- 21633038 TI - A laboratory-based surveillance system for Wuchereria bancrofti in Togo: a practical model for resource-poor settings. AB - One goal of the Global Program to Eliminate Lymphatic Filariasis (GAELF) is interruption of disease transmission through annual mass drug administration (MDA) in areas where LF prevalence is greater than 1%. After MDAs are completed, the World Health Organization (WHO) recommends a period of passive surveillance before final certification of LF elimination is achieved. Guidelines for such a surveillance system have yet to be developed. This paper describes a surveillance system launched in Togo in 2006. The system uses existing laboratories with technicians on call at night who, among other activities, prepare nocturnal thick blood smears for malaria diagnosis that can also be used for LF diagnosis. During its first 2 years (2006-2007), the system provided geographically disperse sampling nationwide, and 1 of 750 people residing in Togo was tested. Over the same period, the system detected two cases of LF, both from areas previously considered non-endemic. This system could be a cost-effective, sustainable model for WHO-mandated passive surveillance after cessation of MDA. PMID- 21633039 TI - Molecular identification of Trichinella papuae from a Thai patient with imported trichinellosis. AB - Previously, we reported the presence of imported trichinellosis in a Thai worker returning from Malaysia, who presented with progressive generalized muscle hypertrophy and weakness after eating wild boar meat. This work analyzed a partial small subunit of a mitochondrial ribosomal RNA gene of Trichinella larvae isolated from the patient. The results showed complete identity with a mitochondrial RNA gene of Trichinella papuae (GenBank accession no. EF517130). This is the first report of imported trichinellosis in Thailand caused by T. papuae. It is possible that T. papuae is widely distributed in the wildlife of Southeast Asia. PMID- 21633040 TI - Diagnosis of Schistosoma haematobium by detection of specific DNA fragments from filtered urine samples. AB - Definitive diagnosis of Schistosoma haematobium infection in adult patients is a clinically important challenge. Chronically infected adults pass few eggs in the urine, which are often missed when current diagnostic methods are used. In the work presented here, we report on an alternative diagnostic method based on presence of the S. haematobium-specific Dra 1, 121 bp repeat fragment in human urine. A novel method of collecting the urine specimens in the field and filtering them through heavy Whatman No. 3 paper was introduced. After drying, the samples remained viable for several months at room temperature. To test the potential use of this method, 89 urine specimens from school children in Kollo District, Niger, were examined. In all, 52 of 89 (58.4%) were positive for hematuria, 4 of 89 (49.4%) were positive for eggs, and 51 of 89 (57.3%) showed parasite-specific DNA. These were compared with 60 filtered urine specimens obtained from random samples of adults from two study sites in Nigeria, one endemic and one non-endemic for S. haematobium. In the 30 patients from the endemic site, all 10 samples with detectable eggs and 7 of the 20 egg-negative samples were DNA positive. It was concluded that the urine filter paper method was sufficiently sensitive to detect low and cryptic infections, that DNA detection was more sensitive than egg detection, and that the filtration method facilitated specimen collection and transport from the field. PMID- 21633041 TI - Human ocular infection with Dirofilaria repens (Railliet and Henry, 1911) in an area endemic for canine dirofilariasis. AB - Dirofilaria repens, which is usually found in canine subcutaneous tissues, is the main causative agent of human dirofilariasis in the Old Word. However, a relationship between animal and human cases of dirofilariasis caused by D. repens in a given area has never been demonstrated. The uneven distribution of D. repens in provinces in Sicily, Italy represented the foundation for this study. We report a human case of ocular infection with D. repens from Trapani Province, where canine dirofilariasis is endemic. The nematode was morphologically and molecularly identified and surgical removal of the parasite was documented. The relationship between the prevalence of D. repens in dogs and the occurrence of human cases of ocular dirofilariasis is discussed on the basis of a review of the historical literature. PMID- 21633042 TI - Molecular characterization of the North American lung fluke Paragonimus kellicotti in Missouri and its development in Mongolian gerbils. AB - Human paragonimiasis is an emerging disease in Missouri. To characterize local parasites, we examined crayfish from three rivers. Metacercaeriae consistent with Paragonimus kellicotti were detected in 69%, 67%, and 37% of crayfish from the Big Piney, Huzzah, and Black Rivers, respectively. Sequencing of the second internal transcribed spacer and other DNA markers confirmed the species identification and the presence of identical parasite sequences in clinical specimens from two human cases. Mongolian gerbils were infected by intraperitoneal injection with 3-8 metacercariae. Most gerbils died 15-49 days post-infection. Necropsies showed pulmonary hemorrhage with necrosis, and flukes as long as 8 mm were recovered from intrathoracic tissues. Western blot analysis using P. kellicotti antigen showed a strong antibody response in gerbils 39 days post-infection. These results demonstrate that P. kellicotti is common in Missouri crayfish. The gerbil model may be useful for research on the pathogenesis, immunology, and treatment of paragonimiasis. PMID- 21633043 TI - Transmission potential of two chimeric Chikungunya vaccine candidates in the urban mosquito vectors, Aedes aegypti and Ae. albopictus. AB - Chikungunya virus (CHIKV) is an emerging, mosquito-borne alphavirus that has caused major epidemics in Africa and Asia. We developed chimeric vaccine candidates using the non-structural protein genes of either Venezuelan equine encephalitis virus (VEEV) attenuated vaccine strain TC-83 or a naturally attenuated strain of eastern equine encephalitis virus (EEEV) and the structural genes of CHIKV. Because the transmission of genetically modified live vaccine strains is undesirable because of the potentially unpredictable evolution of these viruses as well as the potential for reversion, we evaluated the ability of these vaccines to infect the urban CHIKV vectors, Aedes aegypti and Ae. albopictus. Both vaccine candidates exhibited significantly lower infection and dissemination rates compared with the parent alphaviruses. Intrathoracic inoculations indicated that reduced infectivity was mediated by midgut infection barriers in both species. These results indicate a low potential for transmission of these vaccine strains in the event that a vaccinee became viremic. PMID- 21633044 TI - Herpes labialis in patients with Russell's viper bite and acute kidney injury: a single center experience. AB - Snake bite is an important health hazard in tropical countries and is associated with significant morbidity and mortality. Herpes labialis is a common ailment caused by the Herpes simplex virus. There is no published data showing any association between the snake bite and development of Herpes labialis. Here, we present a series of patients who developed Herpes labialis after Russell's viper bite and had acute kidney injury. We attempted to find whether snake bite is an immunosuppressed state and whether it could have pre-disposed the patients to the development of these lesions. PMID- 21633045 TI - The use of positive serological tests as evidence of exposure to Burkholderia pseudomallei. PMID- 21633047 TI - Randomized phase II non-inferiority study (NO16853) of two different doses of capecitabine in combination with docetaxel for locally advanced/metastatic breast cancer. AB - BACKGROUND: This phase II study investigated whether a lower-than-approved dose of capecitabine, plus docetaxel (XT), would improve tolerability versus standard dose XT without compromising efficacy. PATIENTS AND METHODS: Women aged >=18 years with locally advanced/metastatic breast cancer resistant to anthracycline based chemotherapy in the (neo)adjuvant, first- or second-line metastatic setting were eligible. Patients were randomly assigned to receive standard-dose XT (capecitabine 1250 mg/m(2) twice daily, days 1-14; docetaxel 75 mg/m(2), day 1 every 3 weeks) or low-dose XT (capecitabine 825 mg/m(2) twice daily, days 1-14; docetaxel as above). The primary objective was to demonstrate non-inferiority of low-dose to standard-dose XT in terms of progression-free survival (PFS). RESULTS: 470 patients were randomly allocated in a 1 : 1 ratio to standard-dose or low-dose XT. Median PFS was 7.9 versus 5.8 months [hazard ratio 1.16, 95% confidence interval (CI) 0.95-1.43] in the standard-dose and low-dose arms, respectively. The upper limit of the 95% CI was above the predefined non inferiority margin (1.35, P = 0.078). Secondary efficacy end points were consistent with PFS. The frequency and severity of adverse events was similar in both treatment arms. CONCLUSIONS: Non-inferiority of low-dose to standard-dose XT in terms of PFS was not demonstrated; this may be due to regional subgroup effects. PMID- 21633048 TI - Circulating tumor cells, colon cancer and bevacizumab: the meaning of zero. PMID- 21633049 TI - The diagnosis and management of gastric cancer: expert discussion and recommendations from the 12th ESMO/World Congress on Gastrointestinal Cancer, Barcelona, 2010. AB - Well-recognized experts in the field of gastric cancer discussed during the 12th European Society Medical Oncology (ESMO)/World Congress Gastrointestinal Cancer (WCGIC) in Barcelona many important and controversial topics on the diagnosis and management of patients with gastric cancer. This article summarizes the recommendations and expert opinion on gastric cancer. It discusses and reflects on the regional differences in the incidence and care of gastric cancer, the definition of gastro-esophageal junction and its implication for treatment strategies and presents the latest recommendations in the staging and treatment of primary and metastatic gastric cancer. Recognition is given to the need for larger and well-designed clinical trials to answer many open questions. PMID- 21633050 TI - Perfusion CT assessment of tissue hemodynamics following hepatic arterial infusion of increasing doses of angiotensin II in a rabbit liver tumor model. AB - PURPOSE: To investigate the effects of increasing doses of angiotensin II on hepatic hemodynamics in the normal rabbit liver and in hepatic VX2 tumors by using dynamic contrast material-enhanced perfusion computed tomography (CT). MATERIALS AND METHODS: This study was approved by the institutional animal care and use committee. Solitary hepatic VX2 tumors were implanted into 12 rabbits. In each animal, perfusion CT of the liver was performed before (at baseline) and after hepatic arterial infusion of varying doses (0.1-50.0 MUg/mL) of angiotensin II. Images were acquired continuously for 80 seconds after the start of the intravenous contrast material administration. Blood flow (BF), blood volume (BV), mean transit time (MTT), and capillary permeability-surface area product were calculated for the tumor and the adjacent and distant normal liver tissue. Generalized linear mixed models were used to estimate the effects of angiotensin II dose on outcome measures. RESULTS: Angiotensin II infusion increased contrast enhancement of the tumor and distal liver vessels. Tumor BF increased in a dose dependent manner after administration of 0.5-25.0 MUg/mL angiotensin II, but only the 2.5 MUg/mL dose induced a significant increase in tumor BF compared with BF in the adjacent (68.0 vs 26.3 mL/min/100 g, P < .0001) and distant (68.0 vs 28.3 mL/min/100 g, P = .02) normal liver tissue. Tumor BV varied with angiotensin II dose but was greater than the BV of the adjacent and distant liver tissue at only the 2.5 MUg/mL (4.8 vs 3.5 mL/100 g for adjacent liver [P < .0001], 4.8 vs 3.3 mL/100 g for distant liver [P = .0006]) and 10.0 MUg/mL (4.9 vs 4.4 mL/100 g for adjacent liver [P = .007], 4.9 vs 4.3 mL/100 g for distant liver [P = .04]) doses. Tumor MTT was significantly shorter than the adjacent liver tissue MTT at angiotensin II doses of 2.5 MUg/mL (9.7 vs 15.8 sec, P = .001) and 10.0 MUg/mL (5.1 vs 13.2 sec, P = .007) and significantly shorter than the distant liver tissue MTT at 2.5 MUg/mL only (9.7 vs 15.3 sec, P = .0006). The capillary permeability-surface area product for the tumor was higher than that for the adjacent liver tissue at the 2.5 MUg/mL angiotensin II dose only (11.5 vs 8.1 mL/min/100 g, P = .01). CONCLUSION: Perfusion CT enables a mechanistic understanding of angiotensin II infusion in the liver and derivation of the optimal effective dose. The 2.5 MUg/mL angiotensin II dose increases perfusion in hepatic VX2 tumors versus that in adjacent and distant normal liver tissue primarily by constricting normal distal liver vessels and in turn increasing tumor BF and BV. PMID- 21633051 TI - Proximal aneurysmal neck: dynamic ECG-gated CT angiography--conformational pulsatile changes with possible consequences for endograft sizing. AB - PURPOSE: To assess the magnitude of variations in size of the proximal neck of the abdominal aortic aneurysm (AAA) in patients selected to undergo endovascular abdominal aortic aneurysm repair (EVAR) and the potential effect on choice of endograft diameter by using 64-section dynamic electrocardiographically (ECG) gated computed tomographic (CT) angiography. MATERIALS AND METHODS: A prospective single-center study was performed in 40 patients with AAA who underwent both static and dynamic ECG-gated CT angiography. The ethical conduct of the study was approved by the departmental review board, and all patients provided written informed consent. Dynamic ECG-gated data sets were acquired with a low-dose acquisition protocol (100 kV) by using a 0.625-mm-section collimation (40 mL iomeprol [400 mg of iodine per milliliter] versus 80 mL). Pulsatility measurements were taken at suprarenal, juxtarenal, and infrarenal levels within the aneurysmal neck. Manual CT angiographic measurements were performed on modified axial images. On static axial images, one vascular surgeon and one interventional radiologist selected the diameter of the main body of the potential infrarenal stent-graft to implant. The diameter of the main body of the stent-graft selected was compared with the dynamic measurements obtained to calculate the potential relative oversizing performed. RESULTS: A total of 40 patients were enrolled. Significant aortic pulsatility was demonstrated within the aneurysmal neck (mean variation, 9.01% +/- 4.81 [standard deviation]; absolute change, 1.83 mm +/- 1.01; P < .05). When compared with dynamic measurements, the diameter of the endograft selected on the basis of static images would be potentially changed in 12 of 40 (30%) patients. Comparing the minimum or maximum diameter of the aneurysmal neck with the diameter of the endograft selected on the basis of static images, the relative oversizing performed was considered inadequate in seven of 40 (18%) patients. CONCLUSION: Dynamic ECG-gated CT may provide information in regard to pulsatile motion that could change the diameter of the endograft selected on the basis of static imaging measurements. PMID- 21633052 TI - Acute appendicitis in young adults: low- versus standard-radiation-dose contrast enhanced abdominal CT for diagnosis. AB - PURPOSE: To compare low and standard radiation doses in intravenous contrast material-enhanced abdominal computed tomography (CT) for the diagnosis of acute appendicitis in young adults. MATERIALS AND METHODS: The institutional review board approved this retrospective study and waived informed consent. The study included 257 patients (age range, 15-40 years) who underwent CT for suspected appendicitis performed by using a low radiation dose (n = 125) or a standard radiation dose (n = 132). Receiver operating characteristic (ROC) analysis, Fisher exact tests, and Mann-Whitney U tests were used to compare the diagnosis of appendicitis and diagnostic confidence as recorded in prospective CT reports between the two groups. RESULTS: For 55 low-radiation-dose (median dose-length product, 122 mGy . cm) and 44 standard-dose (median dose-length product, 544 mGy . cm) examinations, one of two abdominal radiologists made primary reports that served as final reports. For the remaining examinations, on-call radiologists with differing levels of experience issued preliminary reports and the two abdominal radiologists then provided final reports. In the primary reports, the low- and standard-dose CT groups did not significantly differ in area under the ROC curve (0.96 vs 0.97, P = .76), sensitivity (90% [38 of 42] vs 89% [47 of 53], P > .99), or specificity (92% [76 of 83] vs 94% [74 of 79], P = .74) in the diagnosis of appendicitis. There was also no significant difference between the two groups in the confidence level when diagnosing (P = .71) or excluding (P = .20) appendicitis in the primary reports. Similar results were observed for the final reports. The two dose groups also did not significantly differ in terms of appendiceal visualization, diagnosis of appendiceal perforation, or sensitivity for alternative diagnoses. CONCLUSION: Low-dose CT may have comparable diagnostic performance to standard-dose CT for the diagnosis of appendicitis in young adults. PMID- 21633054 TI - Apparent diffusion coefficient as an MR imaging biomarker of low-risk ductal carcinoma in situ: a pilot study. AB - PURPOSE: To evaluate the potential of apparent diffusion coefficients (ADCs) obtained at quantitative diffusion-weighted magnetic resonance (MR) imaging of the breast as a biomarker of low-grade ductal carcinoma in situ (DCIS). MATERIALS AND METHODS: This retrospective study was approved by an institutional review board, and the requirement to obtain informed consent was waived. Twenty-two women (age range, 36-75 years; mean age, 56.4 years) with pure DCIS (seven with low-grade DCIS, five with intermediate-grade DCIS, and seven with high-grade DCIS) and three with microinvasion underwent breast MR imaging at 1.5 T between January 2008 and November 2010. MR examinations included contrast material enhanced (gadoteridol) T1-weighted imaging and diffusion-weighted MR imaging with b values of 0 and 1000 sec/mm(2). ADC maps were generated. The distributions of the ADCs in regions of interest covering the lesions were compared among the three grades by using linear mixed-model analysis, and the discriminatory power of the lesion minimum ADC was determined with receiver operating characteristic analysis. RESULTS: The mean ADC was 1.42 * 10(-3) mm(2)/sec (95% confidence interval [CI]: 1.31 * 10(-3) mm(2)/sec, 1.54 * 10(-3) mm(2)/sec) for low-grade DCIS, 1.23 * 10(-3) mm(2)/sec (95% CI: 1.10 * 10(-3) mm(2)/sec, 1.36 * 10(-3) mm(2)/sec) for intermediate-grade DCIS, 1.19 * 10(-3) mm(2)/sec (95% CI: 1.08 * 10(-3) mm(2)/sec, 1.30 * 10(-3) mm(2)/sec) for high-grade DCIS, and 2.06 * 10(-3) mm(2)/sec (95% CI: 1.94 * 10(-3) mm(2)/sec, 2.18 * 10(-3) mm(2)/sec) for normal breast tissue. The mean ADCs for high- and intermediate-grade DCIS were significantly lower than that for low-grade DCIS (P < .01 and P = .03, respectively), and the mean ADC for low-grade DCIS was significantly lower than that for normal tissue (P < .001). The lesion minimum ADC for low-grade DCIS was also significantly higher than that for high- and intermediate-grade DCIS (P < .01). A threshold of 1.30 * 10(-3) mm(2)/sec for the minimum ADC in the diagnosis of low-grade DCIS had a specificity of 100% (12 of 12 patients; 95% CI: 73.5%, 100%) and a positive predictive value of 100% (four of four patients; 95% CI: 39.8%, 100%). CONCLUSION: These preliminary results suggest that quantitative diffusion-weighted MR imaging could be used to identify patients with low-grade DCIS with very high specificity. If the results of this study are confirmed, this approach could potentially spare those patients from invasive approaches such as mastectomy or axillary lymph node excision. PMID- 21633053 TI - Potential long-term effects of MDMA on the basal ganglia-thalamocortical circuit: a proton MR spectroscopy and diffusion-tensor imaging study. AB - PURPOSE: To investigate the effects of 3,4-methylenedioxymethamphetamine (MDMA, commonly known as "ecstasy") on the alterations of brain metabolites and anatomic tissue integrity related to the function of the basal ganglia-thalamocortical circuit by using proton magnetic resonance (MR) spectroscopy and diffusion-tensor MR imaging. MATERIALS AND METHODS: This study was approved by a local institutional review board, and written informed consent was obtained from all subjects. Thirty-one long-term (>1 year) MDMA users and 33 healthy subjects were enrolled. Proton MR spectroscopy from the middle frontal cortex and bilateral basal ganglia and whole-brain diffusion-tensor MR imaging were performed with a 3.0-T system. Absolute concentrations of metabolites were computed, and diffusion tensor data were registered to the International Consortium for Brain Mapping template to facilitate voxel-based group comparison. RESULTS: The mean myo inositol level in the basal ganglia of MDMA users (left: 4.55 mmol/L +/- 2.01 [standard deviation], right: 4.48 mmol/L +/- 1.33) was significantly higher than that in control subjects (left: 3.25 mmol/L +/- 1.30, right: 3.31 mmol/L +/- 1.19) (P < .001). Cumulative lifetime MDMA dose showed a positive correlation with the levels of choline-containing compounds (Cho) in the right basal ganglia (r = 0.47, P = .02). MDMA users also showed a significant increase in fractional anisotropy (FA) in the bilateral thalami and significant changes in water diffusion in several regions related to the basal ganglia-thalamocortical circuit as compared with control subjects (P < .05; cluster size, >50 voxels). CONCLUSION: Increased myo-inositol and Cho concentrations in the basal ganglia of MDMA users are suggestive of glial response to degenerating serotonergic functions. The abnormal metabolic changes in the basal ganglia may consequently affect the inhibitory effect of the basal ganglia to the thalamus, as suggested by the increased FA in the thalamus and abnormal changes in water diffusion in the corresponding basal ganglia-thalamocortical circuit. PMID- 21633055 TI - Implications for undergraduate education of two interdisciplinary biological sciences: biochemistry and biophysics. PMID- 21633056 TI - Moving theory into practice: a reflection on teaching a large, introductory biology course for majors. PMID- 21633057 TI - Undergraduate research experiences in biology: alternatives to the apprenticeship model. PMID- 21633058 TI - Recent research in science teaching and learning. PMID- 21633059 TI - Where do new medicines come from? PMID- 21633063 TI - Harnessing technology to improve formative assessment of student conceptions in STEM: forging a national network. AB - Concept inventories, consisting of multiple-choice questions designed around common student misconceptions, are designed to reveal student thinking. However, students often have complex, heterogeneous ideas about scientific concepts. Constructed-response assessments, in which students must create their own answer, may better reveal students' thinking, but are time- and resource-intensive to evaluate. This report describes the initial meeting of a National Science Foundation-funded cross-institutional collaboration of interdisciplinary science, technology, engineering, and mathematics (STEM) education researchers interested in exploring the use of automated text analysis to evaluate constructed-response assessments. Participants at the meeting shared existing work on lexical analysis and concept inventories, participated in technology demonstrations and workshops, and discussed research goals. We are seeking interested collaborators to join our research community. PMID- 21633064 TI - How well do undergraduate research programs promote engagement and success of students? AB - Assessment of undergraduate research (UR) programs using participant surveys has produced a wealth of information about design, implementation, and perceived benefits of UR programs. However, measurement of student participation university wide, and the potential contribution of research experience to student success, also require the study of extrinsic measures. In this essay, institutional data on student credit-hour generation and grade point average (GPA) from the University of Georgia are used to approach these questions. Institutional data provide a measure of annual enrollment in UR classes in diverse disciplines. This operational definition allows accurate and retrospective analysis, but does not measure all modes of engagement in UR. Cumulative GPA is proposed as a quantitative extrinsic measure of student success. Initial results show that extended participation in research for more than a single semester is correlated with an increase in GPA, even after using SAT to control for the initial ability level of the students. While the authors acknowledge that correlation does not prove causality, continued efforts to measure the impact of UR programs on student outcomes using GPA or an alternate extrinsic measure is needed for development of evidence-based programmatic recommendations. PMID- 21633065 TI - Redesigning a large-enrollment introductory biology course. AB - Using an action research model, biology faculty examined, implemented, and evaluated learner-centered instructional strategies to reach the goal of increasing the level of student achievement in the introductory biology course BIO 181: Unity of Life I, which was characterized by both high enrollments and a high DFW rate. Outcomes included the creation and implementation of an assessment tool for biology content knowledge and attitudes, development and implementation of a common syllabus, modification of the course to include learner-centered instructional strategies, and the collection and analysis of data to evaluate the success of the modifications. The redesigned course resulted in greater student success, as measured by grades (reduced %DFW and increased %AB) as well as by achievement in the course assessment tool. In addition, the redesigned course led to increased student satisfaction and greater consistency among different sections. These findings have important implications for both students and institutions, as the significantly lower DFW rate means that fewer students have to retake the course. PMID- 21633066 TI - Increased course structure improves performance in introductory biology. AB - We tested the hypothesis that highly structured course designs, which implement reading quizzes and/or extensive in-class active-learning activities and weekly practice exams, can lower failure rates in an introductory biology course for majors, compared with low-structure course designs that are based on lecturing and a few high-risk assessments. We controlled for 1) instructor effects by analyzing data from quarters when the same instructor taught the course, 2) exam equivalence with new assessments called the Weighted Bloom's Index and Predicted Exam Score, and 3) student equivalence using a regression-based Predicted Grade. We also tested the hypothesis that points from reading quizzes, clicker questions, and other "practice" assessments in highly structured courses inflate grades and confound comparisons with low-structure course designs. We found no evidence that points from active-learning exercises inflate grades or reduce the impact of exams on final grades. When we controlled for variation in student ability, failure rates were lower in a moderately structured course design and were dramatically lower in a highly structured course design. This result supports the hypothesis that active-learning exercises can make students more skilled learners and help bridge the gap between poorly prepared students and their better-prepared peers. PMID- 21633067 TI - Teaching students how to study: a workshop on information processing and self testing helps students learn. AB - We implemented a "how to study" workshop for small groups of students (6-12) for N = 93 consenting students, randomly assigned from a large introductory biology class. The goal of this workshop was to teach students self-regulating techniques with visualization-based exercises as a foundation for learning and critical thinking in two areas: information processing and self-testing. During the workshop, students worked individually or in groups and received immediate feedback on their progress. Here, we describe two individual workshop exercises, report their immediate results, describe students' reactions (based on the workshop instructors' experience and student feedback), and report student performance on workshop-related questions on the final exam. Students rated the workshop activities highly and performed significantly better on workshop-related final exam questions than the control groups. This was the case for both lower- and higher-order thinking questions. Student achievement (i.e., grade point average) was significantly correlated with overall final exam performance but not with workshop outcomes. This long-term (10 wk) retention of a self-testing effect across question levels and student achievement is a promising endorsement for future large-scale implementation and further evaluation of this "how to study" workshop as a study support for introductory biology (and other science) students. PMID- 21633068 TI - Undergraduate science research: a comparison of influences and experiences between premed and non-premed students. AB - Most students participating in science undergraduate research (UR) plan to attend either medical school or graduate school. This study examines possible differences between premed and non-premed students in their influences to do research and expectations of research. Questionnaire responses from 55 premed students and 80 non-premed students were analyzed. No differences existed in the expectations of research between the two groups, but attitudes toward science and intrinsic motivation to learn more about science were significantly higher for non-premed students. Follow-up interviews with 11 of the students, including a case study with one premed student, provided explanation for the observed differences. Premed students, while not motivated to learn more about science, were motivated to help people, which is why most of them are pursuing medicine. They viewed research as a way to help them become doctors and to rule out the possibility of research as a career. Non-premed students participated in research to learn more about a specific science topic and gain experience that may be helpful in graduate school research. The difference in the reasons students want to do UR may be used to tailor UR experiences for students planning to go to graduate school or medical school. PMID- 21633069 TI - Want to improve undergraduate thesis writing? Engage students and their faculty readers in scientific peer review. AB - One of the best opportunities that undergraduates have to learn to write like a scientist is to write a thesis after participating in faculty-mentored undergraduate research. But developing writing skills doesn't happen automatically, and there are significant challenges associated with offering writing courses and with individualized mentoring. We present a hybrid model in which students have the structural support of a course plus the personalized benefits of working one-on-one with faculty. To optimize these one-on-one interactions, the course uses BioTAP, the Biology Thesis Assessment Protocol, to structure engagement in scientific peer review. By assessing theses written by students who took this course and comparable students who did not, we found that our approach not only improved student writing but also helped faculty members across the department--not only those teaching the course--to work more effectively and efficiently with student writers. Students who enrolled in this course were more likely to earn highest honors than students who only worked one on-one with faculty. Further, students in the course scored significantly better on all higher-order writing and critical-thinking skills assessed. PMID- 21633070 TI - Can genetics and genomics nursing competencies be successfully taught in a prenursing microbiology course? AB - In recognition of the entry into the era of personalized medicine, a new set of genetics and genomics competencies for nurses was introduced in 2006. Since then, there have been a number of reports about the critical importance of these competencies for nursing practices and about the challenges of addressing these competencies in the preservice (basic science) nursing curriculum. At least one suggestion has been made to infuse genetics and genomics throughout the basic science curriculum for prenursing students. Based on this call and a review of the competencies, this study sought to assess the impact of incorporation of genetics and genomics content into a prenursing microbiology course. Broadly, two areas that address the competencies were incorporated into the course: 1) the biological basis and implications of genetic diversity and 2) the technological aspects of assessing genetic diversity in bacteria and viruses. These areas address how genetics and genomics contribute to healthcare, including diagnostics and selection of treatment. Analysis of learning gains suggests that genetics and genomics content can be learned as effectively as microbiology content in this setting. Future studies are needed to explore the most effective ways to introduce genetics and genomics technology into the prenursing curriculum. PMID- 21633071 TI - Using digital images of the zebra finch song system as a tool to teach organizational effects of steroid hormones: a free downloadable module. AB - Zebra finch song behavior is sexually dimorphic: males sing and females do not. The neural system underlying this behavior is sexually dimorphic, and this sex difference is easy to quantify. During development, the zebra finch song system can be altered by steroid hormones, specifically estradiol, which actually masculinizes it. Because of the ease of quantification and experimental manipulation, the zebra finch song system has great potential for use in undergraduate labs. Unfortunately, the underlying costs prohibit use of this system in undergraduate labs. Further, the time required to perform a developmental study renders such undertakings unrealistic within a single academic term. We have overcome these barriers by creating digital tools, including an image library of song nuclei from zebra finch brains. Students using this library replicate and extend a published experiment examining the dose of estradiol required to masculinize the female zebra finch brain. We have used this library for several terms, and students not only obtain significant experimental results but also make gains in understanding content, experimental controls, and inferential statistics (analysis of variance and post hoc tests). We have provided free access to these digital tools at the following website: http://mdcune.psych.ucla.edu/modules/birdsong. PMID- 21633072 TI - Associations of growth trajectories in infancy and early childhood with later childhood outcomes. AB - BACKGROUND: Weight and length at birth (which represent fetal growth) and weight and length or height gain during childhood (which potentially represent catch-up growth) may be related to later health outcomes. However, methods for the assessment of such relations are complex and underdeveloped. OBJECTIVES: We aimed to describe childhood weight and length or height trajectories and to relate these to later outcomes by using rash at age 6.5 y as an example. DESIGN: The data came from a prospective cohort study in Belarus in 10,494 children born in 31 hospitals that participated in a cluster randomized trial of breastfeeding promotion. Weight and length or height were measured at birth, at scheduled clinic visits up to 1 y, and at 6.5 y; intermediate measures were obtained from routine child health records. Linear spline multilevel models for weight and length or height were used to estimate each child's deviance from average birth weight, birth length, weight, and length or height gain velocity in each time period. Logistic regression was used to relate the outcome (parental report of rash at 6.5 y) to these weight and length or height estimates. RESULTS: The best fitting splines for length or height and weight had knots at 3 and 12 mo, with another knot at 34 mo for height. The only relation between weight and length or height and reported rash was a positive association with weight gain velocity between 12 and 34 mo (odds ratio per SD increase in weight gain velocity: 1.11; 95% CI: 1.01, 1.22). CONCLUSION: Advantages of multilevel models include no restriction to measures at arbitrary times or to individuals with complete data and allowance for measurement error. This trial was registered at isrctn.org as ISRCTN37687716. PMID- 21633073 TI - Early determinants of type 1 diabetes: experience from the BABYDIAB and BABYDIET studies. AB - Type 1 diabetes is an immune-mediated disorder that results from progressive destruction of the islet beta cells. A genetic susceptibility for the development of islet autoimmunity and type 1 diabetes is well documented, and an environmental influence is assumed. Prospective studies from birth have shown that islet autoimmunity occurs very early in life, which implies that fetal or postnatal environmental factors may program the development of islet autoimmunity. In this overview, results from the BABYDIAB study, a prospective study from birth on the natural history of type 1 diabetes, and the BABYDIET study, a dietary intervention study, are discussed with a focus on the role of a diabetes environment in utero and the role of early gluten exposure on islet autoimmunity risk in children. PMID- 21633074 TI - Effects of supplemented isoenergetic diets differing in cereal fiber and protein content on insulin sensitivity in overweight humans. AB - BACKGROUND: Despite their beneficial effects on weight loss and blood lipids, high-protein (HP) diets have been shown to increase insulin resistance and diabetes risk, whereas high-cereal-fiber (HCF) diets have shown the opposite effects on these outcomes. OBJECTIVE: We compared the effects of isoenergetic HP and HCF diets and a diet with moderate increases in both cereal fibers and dietary protein (Mix diet) on insulin sensitivity, as measured by using euglycemic-hyperinsulinemic clamps with infusion of [6,6-(2)H(2)]glucose. DESIGN: We randomly assigned 111 overweight adults with features of the metabolic syndrome to 1 of 4 two-phased, 18-wk isoenergetic diets by group-matching. Per 3 d food protocols, the percentages of energy derived from protein and carbohydrates and the intake of cereal fiber per day, respectively, were as follows-after 6 wk: 17%, 52%, and 14 g (control); 17%, 52%, and 43 g (HCF); 28%, 43%, and 13 g (HP); 23%, 44%, and 26 g (Mix); after 18 wk: 17%, 51%, and 15 g (control); 17%, 51%, and 41 g (HCF); 26%, 45%, and 14 g (HP); and 22%, 46%, and 26 g (Mix). Eighty-four participants completed the study successfully and were included in the final analyses. Adherence was supported by the provision of tailored dietary supplements twice daily in all groups. RESULTS: Insulin sensitivity expressed as an M value was 25% higher after 6 wk of the HCF diet than after 6 wk of the HP diet (subgroup analysis: 4.61 +/- 0.38 compared with 3.71 +/- 0.36 mg . kg(-1) . min(-1), P = 0.008; treatment * time interaction: P = 0.005). Effects were attenuated after 18 wk (treatment * time interaction: P = 0.054), which was likely explained by lower adherence to the HP diet. HP intake was associated with a tendency to increased protein expression in adipose tissue of the translation initiation factor serine-kinase-6-1, which is known to mediate amino acid-induced insulin resistance. Biomarkers of protein intake indicated interference of cereal fibers with dietary protein absorption. CONCLUSION: Greater changes in insulin sensitivity after intake of an isoenergetic HCF than after intake of an HP diet might help to explain the diverse effects of these diets on diabetes risk. This trial is registered at clinicaltrials.gov as NCT00579657. PMID- 21633075 TI - The hybrid science of diet, microbes, and metabolic health. PMID- 21633076 TI - The risk of kidney stone formation: the form of calcium matters. PMID- 21633077 TI - Air bubble contact with endothelial cells in vitro induces calcium influx and IP3 dependent release of calcium stores. AB - Gas embolism is a serious complication of decompression events and clinical procedures, but the mechanism of resulting injury remains unclear. Previous work has demonstrated that contact between air microbubbles and endothelial cells causes a rapid intracellular calcium transient and can lead to cell death. Here we examined the mechanism responsible for the calcium rise. Single air microbubbles (50-150 MUm), trapped at the tip of a micropipette, were micromanipulated into contact with individual human umbilical vein endothelial cells (HUVECs) loaded with Fluo-4 (a fluorescent calcium indicator). Changes in intracellular calcium were then recorded via epifluorescence microscopy. First, we confirmed that HUVECs rapidly respond to air bubble contact with a calcium transient. Next, we examined the involvement of extracellular calcium influx by conducting experiments in low calcium buffer, which markedly attenuated the response, or by pretreating cells with stretch-activated channel blockers (gadolinium chloride or ruthenium red), which abolished the response. Finally, we tested the role of intracellular calcium release by pretreating cells with an inositol 1,4,5-trisphosphate (IP3) receptor blocker (xestospongin C) or phospholipase C inhibitor (neomycin sulfate), which eliminated the response in 64% and 67% of cases, respectively. Collectively, our results lead us to conclude that air bubble contact with endothelial cells causes an influx of calcium through a stretch-activated channel, such as a transient receptor potential vanilloid family member, triggering the release of calcium from intracellular stores via the IP3 pathway. PMID- 21633078 TI - Novel phosphorylation of aquaporin-5 at its threonine 259 through cAMP signaling in salivary gland cells. AB - Aquaporin-5 (AQP5), a water channel, plays key roles in salivary secretion. The novel phosphorylation of AQP5 was investigated by using human salivary gland (HSG) cells and mouse salivary glands. In the HSG cells stably transfected with a wild-type mouse AQP5 construct, a protein band immunoreactive with antibody against phosphorylated PKA substrate was detected in the AQP5 immunoprecipitated sample, and its intensity was enhanced by short-term treatment of the cells with 8-bromo-cAMP, forskolin, or phorbol 12-myristate 13-acetate, but not by that with A23187 calcium ionophore. Such enhancement was inhibited in the presence of H-89, a PKA inhibitor. An AQP5 mutant (AQP5-T259A) expressed by transfection of HSG cells was not recognized by anti-phosphorylated PKA substrate antibody, even when the cells were stimulated with the protein kinase activators. Immunoblotting and immunofluorescence studies using a specific antibody detecting AQP5 phosphorylated at its Thr259 demonstrated that AQP5 was rapidly and transiently phosphorylated at the apical membrane of acinar cells in the submandibular and parotid glands after administration of isoproterenol, but not pilocarpine. Furthermore, both AQP5 and AQP5-T259A were constitutively localized at the plasma membrane in HSG cells under the resting and forskolin-stimulated conditions. These results suggest that AQP5 is phosphorylated at its Thr259 by PKA through cAMP, but not Ca(2+), signaling pathways, and that this phosphorylation does not contribute to AQP5 trafficking in the salivary gland cells. PMID- 21633079 TI - In dialyzed squid axons oxidative stress inhibits the Na+/Ca2+ exchanger by impairing the Cai2+-regulatory site. AB - The Na(+)/Ca(2+) exchanger, a major mechanism by which cells extrude calcium, is involved in several physiological and physiopathological interactions. In this work we have used the dialyzed squid giant axon to study the effects of two oxidants, SIN-1-buffered peroxynitrite and hydrogen peroxide (H(2)O(2)), on the Na(+)/Ca(2+) exchanger in the absence and presence of MgATP upregulation. The results show that oxidative stress induced by peroxynitrite and hydrogen peroxide inhibits the Na(+)/Ca(2+) exchanger by impairing the intracellular Ca(2+) (Ca(i)(2+))-regulatory sites, leaving unharmed the intracellular Na(+)- and Ca(2+)-transporting sites. This effect is efficiently counteracted by the presence of MgATP and by intracellular alkalinization, conditions that also protect H(i)(+) and (H(i)(+) + Na(i)(+)) inhibition of Ca(i)(2+)-regulatory sites. In addition, 1 mM intracellular EGTA reduces oxidant inhibition. However, once the effects of oxidants are installed they cannot be reversed by either MgATP or EGTA. These results have significant implications regarding the role of the Na(+)/Ca(2+) exchanger in response to pathological conditions leading to tissue ischemia-reperfusion and anoxia/reoxygenation; they concur with a marked reduction in ATP concentration, an increase in oxidant production, and a rise in intracellular Ca(2+) concentration that seems to be the main factor responsible for cell damage. PMID- 21633080 TI - Detecting Ca2+ sparks on stationary and varying baselines. AB - Studies concerning the physiological significance of Ca(2+) sparks often depend on the detection and measurement of large populations of events in noisy microscopy images. Automated detection methods have been developed to quickly and objectively distinguish potential sparks from noise artifacts. However, previously described algorithms are not suited to the reliable detection of sparks in images where the local baseline fluorescence and noise properties can vary significantly, and risk introducing additional bias when applied to such data sets. Here, we describe a new, conceptually straightforward approach to spark detection in linescans that addresses this issue by combining variance stabilization with local baseline subtraction. We also show that in addition to greatly increasing the range of images in which sparks can be automatically detected, the use of a more accurate noise model enables our algorithm to achieve similar detection sensitivities with fewer false positives than previous approaches when applied both to synthetic and experimental data sets. We propose, therefore, that it might be a useful tool for improving the reliability and objectivity of spark analysis in general, and describe how it might be further optimized for specific applications. PMID- 21633081 TI - Quantifying immunogold localization on electron microscopic thin sections: a compendium of new approaches for plant cell biologists. AB - A review is presented of recently developed methods for quantifying electron microscopical thin sections on which colloidal gold-labelled markers are used to identify and localize interesting molecules. These efficient methods rely on sound principles of random sampling, event counting, and statistical evaluation. Distributions of immunogold particles across cellular compartments can be compared within and between experimental groups. They can also be used to test for co-localization in multilabelling studies involving two or more sizes of gold particle. To test for preferential labelling of compartments, observed and expected gold particle distributions are compared by chi(2) analysis. Efficient estimators of gold labelling intensity [labelling density (LD) and/or relative labelling index (RLI)] are used to analyse volume-occupying compartments (e.g. Golgi vesicles) and/or surface-occupying compartments (e.g. cell membranes). Compartment size is estimated by counting chance events after randomly superimposing test lattices of points and/or line probes. RLI=1 when there is random labelling and RLI >1 when there is preferential labelling. Between-group comparisons do not require information about compartment size but, instead, raw gold particle counts in different groups are compared by combining chi(2) and contingency table analyses. These tests may also be used to assess co distribution of different sized gold particles in compartments. Testing for co labelling involves identifying sets of compartmental profiles that are unlabelled and labelled for one or both of two gold marker sizes. Numbers of profiles in each labelling set are compared by contingency table analysis and chi(2) analysis or Fisher's exact probability test. The various methods are illustrated with worked examples based on empirical and synthetic data and will be of practical benefit to those applying single or multiple immunogold labelling in their research. PMID- 21633082 TI - The role of epigenetic processes in controlling flowering time in plants exposed to stress. AB - Plants interact with their environment by modifying gene expression patterns. One mechanism for this interaction involves epigenetic modifications that affect a number of aspects of plant growth and development. Thus, the epigenome is highly dynamic in response to environmental cues and developmental changes. Flowering is controlled by a set of genes that are affected by environmental conditions through an alteration in their expression pattern. This ensures the production of flowers even when plants are growing under adverse conditions, and thereby enhances transgenerational seed production. In this review recent findings on the epigenetic changes associated with flowering in Arabidopsis thaliana grown under abiotic stress conditions such as cold, drought, and high salinity are discussed. These epigenetic modifications include DNA methylation, histone modifications, and the production of micro RNAs (miRNAs) that mediate epigenetic modifications. The roles played by the phytohormones abscisic acid (ABA) and auxin in chromatin remodelling are also discussed. It is shown that there is a crucial relationship between the epigenetic modifications associated with floral initiation and development and modifications associated with stress tolerance. This relationship is demonstrated by the common epigenetic pathways through which plants control both flowering and stress tolerance, and can be used to identify new epigenomic players. PMID- 21633083 TI - Border sequences of Medicago truncatula CLE36 are specifically cleaved by endoproteases common to the extracellular fluids of Medicago and soybean. AB - CLE (CLAVATA3/ESR-related) peptides are developmental regulators that are secreted into the apoplast. Little is known about the role of the sequences that flank CLE peptides in terms of their biological activity or how they are targeted by proteases that are known to liberate the final active CLE peptides from their precursor sequences. The biological activity of Medicago truncatula CLE36, which possesses broadly conserved border sequences flanking the putative final active CLE36 peptide product, was assessed. Using in vitro root growth assays and an in vitro root and callus formation assay it is shown that CLE36 peptides of different lengths possess differential biological activities. Using mass spectrometry, Glycine max and Medicago extracellular fluids were each shown to possess an endoproteolytic activity that recognizes and cleaves at border sequences in a synthetic 31 amino acid CLE36 'propeptide bait' to liberate biologically active peptide products. Inhibitor studies suggest that a subtilisin, in combination with a carboxypeptidase, liberated and trimmed CLE36, respectively, to form biologically relevant 11-15 amino acid cleavage products. The 15 amino acid cleavage product is more biologically potent on Arabidopsis than shorter or longer CLE peptides. In situ hybridization shows that the soybean orthologue of CLE36 (GmCLE34) is expressed in the provascular tissue. The results suggest that secreted subtilisins can specifically recognize the border sequences of CLE36 propeptides and liberate biologically active cleavage products. These secreted proteases may affect the stability and biological activity of CLE peptides in the apoplast or be involved in CLE36 processing. PMID- 21633084 TI - Functional characterization of a class III acid endochitinase from the traps of the carnivorous pitcher plant genus, Nepenthes. AB - Carnivory in plants is an adaptation strategy to nutrient-poor environments and soils. Carnivorous plants obtain some additional mineral nutrients by trapping and digesting prey; the genus Nepenthes is helped by its specialized pitcher traps. To make the nutrients available, the caught prey needs to be digested, a process that requires the concerted activity of several hydrolytic enzymes. To identify and investigate the various enzymes involved in this process, fluid from Nepenthes traps has been analysed in detail. In this study, a novel type of Nepenthes endochitinase was identified in the digestion fluid of closed pitchers. The encoding endochitinase genes have been cloned from eight different Nepenthes species. Among these, the deduced amino acid sequence similarity was at least 94.9%. The corresponding cDNA from N. rafflesiana was heterologously expressed, and the purified protein, NrChit1, was biochemically characterized. The enzyme, classified as a class III acid endochitinase belonging to family 18 of the glycoside hydrolases, is secreted into the pitcher fluid very probably due to the presence of an N-terminal signal peptide. Transcriptome analyses using real-time PCR indicated that the presence of prey in the pitcher up-regulates the endochitinase gene not only in the glands, which are responsible for enzyme secretion, but at an even higher level, in the glands' surrounding tissue. These results suggest that in the pitchers' tissues, the endochitinase as well as other proteins from the pitcher fluid might fulfil a different, primary function as pathogenesis-related proteins. PMID- 21633085 TI - Polyamine-induced modulation of genes involved in ethylene biosynthesis and signalling pathways and nitric oxide production during olive mature fruit abscission. AB - After fruit ripening, many fruit-tree species undergo massive natural fruit abscission. Olive (Olea europaea L.) is a stone-fruit with cultivars such as Picual (PIC) and Arbequina (ARB) which differ in mature fruit abscission potential. Ethylene (ET) is associated with abscission, but its role during mature fruit abscission remains largely uncharacterized. The present study investigates the possible roles of ET and polyamine (PA) during mature fruit abscission by modulating genes involved in the ET signalling and biosynthesis pathways in the abscission zone (AZ) of both cultivars. Five ET-related genes (OeACS2, OeACO2, OeCTR1, OeERS1, and OeEIL2) were isolated in the AZ and adjacent cells (AZ-AC), and their expression in various olive organs and during mature fruit abscission, in relation to interactions between ET and PA and the expression induction of these genes, was determined. OeACS2, OeACO2, and OeEIL2 were found to be the only genes that were up-regulated in association with mature fruit abscission. Using the inhibition of ET and PA biosynthesis, it is demonstrated that OeACS2 and OeEIL2 expression are under the negative control of PA while ET induces their expression in AZ-AC. Furthermore, mature fruit abscission depressed nitric oxide (NO) production present mainly in the epidermal cells and xylem of the AZ. Also, NO production was differentially responsive to ET, PA, and different inhibitors. Taken together, the results indicate that PA dependent ET signalling and biosynthesis pathways participate, at least partially, during mature fruit abscission, and that endogenous NO and 1 aminocyclopropane-1-carboxylic acid maintain an inverse correlation, suggesting an antagonistic action of NO and ET in abscission signalling. PMID- 21633086 TI - Discontinuing prophylactic transfusions increases the risk of silent brain infarction in children with sickle cell disease: data from STOP II. AB - In the STOP II trial, discontinuation of prophylactic transfusions in high risk children with sickle cell disease (SCD) resulted in a high rate of reversion to abnormal blood-flow velocities on transcranial Doppler (TCD) ultrasonography and strokes. We analyzed data from STOP II to determine the effect of discontinuing transfusions on the development or progression of silent brain infarcts on magnetic resonance imaging (MRI). At study entry, 21 of 79 (27%) patients had evidence of silent infarcts. There were no statistically significant differences in baseline characteristics between patients with normal brain MRI or silent infarcts at study entry. At study end, 3 of 37 (8.1%) patients in the continued transfusion group developed new brain MRI lesions compared with 11 of 40 (27.5%) in the transfusion-halted group (P = .03). The total number of lesions remained essentially unchanged decreasing from 25 to 24 in the continued-transfusion group while increasing from 27 to 45 in transfusion-halted patients. Thus, discontinuation of transfusions in children with SCD and abnormal TCD who revert to low-risk increases the risk of silent brain infarction. Together with data from STOP, these findings demonstrate that transfusions prevent the development of silent infarcts in patients with SCD and abnormal TCD but normal MRA. PMID- 21633087 TI - Influence of immunosuppressive treatment on risk of recurrent malignancy after allogeneic hematopoietic cell transplantation. AB - This study was conducted to elucidate the influence of immunosuppressive treatment (IST) and GVHD on risk of recurrent malignancy after allogeneic hematopoietic cell transplantation (HCT). The study cohort included 2656 patients who received allogeneic HCT after high-intensity conditioning regimens for treatment of hematologic malignancies. Rates and hazard ratios of relapse and mortality were analyzed according to GVHD and IST as time-varying covariates. Adjusted Cox analyses showed that acute and chronic GVHD were both associated with statistically similar reductions in risk of relapse beyond 18 months after HCT but not during the first 18 months. In patients with GVHD, resolution of GVHD followed by withdrawal of IST was not associated with a subsequent increase in risk of relapse. In patients without GVHD, withdrawal of IST was associated with a reduced risk of relapse during the first 18 months, but the risk of subsequent relapse remained considerably higher than in patients with GVHD. In summary, the association of GVHD with risk of relapse changes over time after HCT. In patients without GVHD, early withdrawal of IST might help to prevent relapse during the first 18 months, but other interventions would be needed to prevent relapse at later time points. PMID- 21633088 TI - Differentiation of human peripheral blood Vdelta1+ T cells expressing the natural cytotoxicity receptor NKp30 for recognition of lymphoid leukemia cells. AB - The success of cancer immunotherapy depends on productive tumor cell recognition by killer lymphocytes. gammadelta T cells are a population of innate-like lymphocytes endowed with strong, MHC-unrestricted cytotoxicity against tumor cells. This notwithstanding, we recently showed that a large proportion of human hematologic tumors is resistant to gammadelta peripheral blood lymphocytes (PBLs) activated with specific agonists to the highly prevalent Vgamma9Vdelta2 TCR. Although this probably constitutes an important limitation to current gammadelta T cell-mediated immunotherapy strategies, we describe here the differentiation of a novel subset of Vdelta2(-) Vdelta1(+) PBLs expressing natural cytotoxicity receptors (NCRs) that directly mediate killing of leukemia cell lines and chronic lymphocytic leukemia patient neoplastic cells. We show that Vdelta1(+) T cells can be selectively induced to express NKp30, NKp44 and NKp46, through a process that requires functional phosphatidylinositol 3-kinase (PI-3K)/AKT signaling on stimulation with gamma(c) cytokines and TCR agonists. The stable expression of NCRs is associated with high levels of granzyme B and enhanced cytotoxicity against lymphoid leukemia cells. Specific gain-of-function and loss-of-function experiments demonstrated that NKp30 makes the most important contribution to TCR independent leukemia cell recognition. Thus, NKp30(+) Vdelta1(+) T cells constitute a novel, inducible and specialized killer lymphocyte population with high potential for immunotherapy of human cancer. PMID- 21633089 TI - miRNA expression in diffuse large B-cell lymphoma treated with chemoimmunotherapy. AB - Diffuse large B-cell lymphoma (DLBCL) prognostication requires additional biologic markers. miRNAs may constitute markers for cancer diagnosis, outcome, or therapy response. In the present study, we analyzed the miRNA expression profile in a retrospective multicenter series of 258 DLBCL patients uniformly treated with chemoimmunotherapy. Findings were correlated with overall survival (OS) and progression-free survival (PFS). miRNA and gene-expression profiles were studied using microarrays in an initial set of 36 cases. A selection of miRNAs associated with either DLBCL molecular subtypes (GCB/ABC) or clinical outcome were studied by multiplex RT-PCR in a test group of 240 cases with available formalin-fixed, paraffin-embedded (FFPE) diagnostic samples. The samples were divided into a training set (123 patients) and used to derive miRNA-based and combined (with IPI score) Cox regression models in an independent validation series (117 patients). Our model based on miRNA expression predicts OS and PFS and improves upon the predictions based on clinical variables. Combined models with IPI score identified a high-risk group of patients with a 2-year OS and a PFS probability of < 50%. In summary, a precise miRNA signature is associated with poor clinical outcome in chemoimmunotherapy-treated DLBCL patients. This information improves upon IPI-based predictions and identifies a subgroup of candidate patients for alternative therapeutic regimens. PMID- 21633090 TI - Irgm1 protects hematopoietic stem cells by negative regulation of IFN signaling. AB - The IFN-inducible immunity-related p47 GTPase Irgm1 has been linked to Crohn disease as well as susceptibility to tuberculosis. Previously we demonstrated that HSC quiescence and function are aberrant in mice lacking Irgm1. To investigate the molecular basis for these defects, we conducted microarray expression profiling of Irgm1-deficient HSCs. Cell-cycle and IFN-response genes are up-regulated in Irgm1(-/-) HSCs, consistent with dysregulated IFN signaling. To test the hypothesis that Irgm1 normally down-regulates IFN signaling in HSCs, we generated Irgm1(-/-)Ifngr1(-/-) and Irgm1(-/-)Stat1(-/-) double-knockout animals. Strikingly, hyperproliferation, self-renewal, and autophagy defects in Irgm1(-/-) HSCs were normalized in double-knockout animals. These defects were also abolished in Irgm1(-/-)Irgm3(-/-) double-knockout animals, indicating that Irgm1 may regulate Irgm3 activity. Furthermore, the number of HSCs was reduced in aged Irgm1(-/-) animals, suggesting that negative feedback inhibition of IFN signaling by Irgm1 is necessary to prevent hyperproliferation and depletion of the stem cell compartment. Collectively, our results indicate that Irgm1 is a powerful negative regulator of IFN-dependent stimulation in HSCs, with an essential role in preserving HSC number and function. The deleterious effects of excessive IFN signaling may explain how hematologic abnormalities arise in patients with inflammatory conditions. PMID- 21633091 TI - Design and implementation of a hybrid genetic algorithm and artificial neural network system for predicting the sizes of unerupted canines and premolars. AB - The aim of this study was to develop a novel hybrid genetic algorithm and artificial neural network (GA-ANN) system for predicting the sizes of unerupted canines and premolars during the mixed dentition period. This study was performed on 106 untreated subjects (52 girls, 54 boys, aged 13-15 years). Data were obtained from dental cast measurements. A hybrid GA-ANN algorithm was developed to find the best reference teeth and the most accurate mapping function. Based on a regression analysis, the strongest correlation was observed between the sum of the mesiodistal widths of the mandibular canines and premolars and the mesiodistal widths of the mandibular first molars and incisors (r = 0.697). In the maxilla, the highest correlation was observed between the sum of the mesiodistal widths of the canines and premolars and the mesiodistal widths of the mandibular first molars and maxillary central incisors (0.742). The hybrid GA-ANN algorithm selected the mandibular first molars and incisors and the maxillary central incisors as the reference teeth for predicting the sum of the mesiodistal widths of the canines and premolars. The prediction error rates and maximum rates of over/underestimation using the hybrid GA-ANN algorithm were smaller than those using linear regression analyses. PMID- 21633092 TI - Cost-effectiveness of colorectal cancer screening. AB - Colorectal cancer is an important public health problem. Several screening methods have been shown to be effective in reducing colorectal cancer mortality. The objective of this review was to assess the cost-effectiveness of the different colorectal cancer screening methods and to determine the preferred method from a cost-effectiveness point of view. Five databases (MEDLINE, EMBASE, the Cost-Effectiveness Analysis Registry, the British National Health Service Economic Evaluation Database, and the lists of technology assessments of the Centers for Medicare and Medicaid Services) were searched for cost-effectiveness analyses published in English between January 1993 and December 2009. Fifty-five publications relating to 32 unique cost-effectiveness models were identified. All studies found that colorectal cancer screening was cost-effective or even cost saving compared with no screening. However, the studies disagreed as to which screening method was most effective or had the best incremental cost effectiveness ratio for a given willingness to pay per life-year gained. There was agreement among studies that the newly developed screening tests of stool DNA testing, computed tomographic colonography, and capsule endoscopy were not yet cost-effective compared with the established screening options. PMID- 21633094 TI - Vasopressin secretion by hypertonic saline infusion during hemodialysis: effect of cardiopulmonary recirculation. AB - BACKGROUND: Intradialytic hypotension is the most common and severe acute complication of hemodialysis therapy. In our previous study, infusion of 20 mL of 10% saline into the venous line of a dialyzer increased blood pressure during dialysis hypotension by stimulating arginine vasopressin (AVP) secretion, independent of its effect on plasma volume (PV). This study examines the mechanism by which a small amount of hypertonic solution stimulates AVP secretion. METHODS: Hemodialysis patients were infused with 20 mL of 2.5 M saline (100 mOsm) over 5 (Protocol 1) or 2 min (Protocol 2) or with isotonic saline (Protocol 3) into the venous line. RESULTS: Arterial plasma osmolality (Posm) increased by 28.1 and 16.0 (P < 0.0001), while peripheral venous Posm increased by only 8.6 and 8.9 mOsm/kg H(2)O (P < 0.001) in Protocols 2 and 1, respectively. Plasma AVP (P(AVP)) increased significantly by 18.6 and 5.6 pg/mL, PV by 7.2 and 5.5% and mean arterial pressure (MAP) by 15.0 and 7.2 mmHg in Protocols 2 and 1, respectively. Thus, there were large differences in Posm between arterial and peripheral venous blood; osmolar gap, P(AVP) and MAP increased in proportion to the infusion rate. Isotonic saline (30.8 mOsm) infusion increased PV by 8.7% and MAP by 7.2 mmHg. CONCLUSION: Our results indicate that by a mechanism similar to cardiopulmonary recirculation, hypertonic saline infusion caused a striking increase in arterial Posm that enhanced AVP secretion and raised blood pressure. The effect of hypertonic saline on PV was less than one-third of isotonic saline under similar osmolar loads. PMID- 21633093 TI - A role for miR-296 in the regulation of lipoapoptosis by targeting PUMA. AB - Saturated free fatty acids (FFA) induce hepatocyte lipoapoptosis, a key mediator of liver injury in nonalcoholic fatty liver disease (NAFLD). Lipoapoptosis involves the upregulation of the BH3-only protein PUMA, a potent pro-apoptotic protein. Given that dysregulation of hepatic microRNA expression has been observed in NAFLD, we examined the role of miRNA in regulating PUMA expression during lipotoxicity. By in silico analysis, we identified two putative binding sites for miR-296-5p within the 3' untranslated region (UTR) of PUMA mRNA. Enforced miR-296-5p levels efficiently reduced PUMA protein expression in Huh-7 cells, while antagonism of miR-296-5p function increased PUMA cellular levels. Reporter gene assays identified PUMA 3'UTR as a direct target of miR-296-5p. The saturated FFA, palmitate, repressed miR-296-5p expression; and Huh-7 cells were sensitized to palmitate-induced lipotoxicity by antagonism of miR-296-5p function using a targeted locked nucleic acid (LNA). Finally, miR-296-5p was reduced in liver samples from nonalcoholic steatohepatitis (NASH) patients compared with patients with simple steatosis (SS) or controls. Also miR-296-5p levels inversely varied with PUMA mRNA levels in human liver specimens. Our results implicate miR 296-5p in the regulation of PUMA expression during hepatic lipoapoptosis. We speculate that enhancement of miR-296-5p expression may represent a novel approach to minimize apoptotic damage in human fatty liver diseases. PMID- 21633095 TI - High glucose-induced hypertrophy of mesangial cells is reversed by connexin43 overexpression via PTEN/Akt/mTOR signaling. AB - BACKGROUND: Hypertrophy of glomerular mesangial cells (GMC) is one of the earliest pathological abnormalities in diabetic nephropathy, which correlates with eventual glomerulosclerosis. We have previously proved that this hypertrophy is mediated by downregulation of connexin43 (Cx43) and dysfunction of gap junctional intercellular communication, but the mechanism involved is still unclear. This study aims to investigate whether PTEN/Akt/mammalian target of rapamycin (mTOR) was involved as the downstream molecular signaling of Cx43 in regulating high glucose-induced GMC hypertrophy. METHODS: GMC were isolated from male Wistar rats at the age of 3 months. Gene transfer technique was used to upregulate Cx43 in GMC, which was validated by western blot and immunofluorescent staining. Forward scatter of flow cytometry and total protein/cell number were examined to testify GMC hypertrophy induced by high glucose (30 mM) and the influence of Cx43 overexpression; western blot was performed to demonstrate the changes of Cx43 and signal protein level and flow cytometry and MTT test were carried out to check cell cycle and proliferation rate, respectively. RESULTS: GMC exposed to a high concentration of glucose presented decreased Cx43, inhibited PTEN, triggered Akt phosphorylation and activated downstream mTOR, leading to stagnancy of cell cycle, decline of proliferation rate and occurrence of hypertrophy. Cx43 overexpression could prevent PTEN inhibition, Akt and mTOR phosphorylation, resulting in restoration of cell cycle and proliferation ability and reversion of GMC hypertrophy. GMC with Cx43 inhibition showed similar PTEN/Akt/mTOR change as stimulated by high glucose. CONCLUSION: PTEN/Akt/mTOR signaling stimulated by high concentration of glucose is regulated by Cx43 overexpression, which unveils part of the molecular mechanism of Cx43 in regulating hyperglycemia-induced hypertrophy. PMID- 21633096 TI - Human peritoneal mesothelial cells respond to bacterial ligands through a specific subset of Toll-like receptors. AB - BACKGROUND: Bacterial infection remains a major cause of morbidity and mortality in peritoneal dialysis (PD) patients worldwide. Previous studies have identified a key role for mesothelial cells, lining the peritoneal cavity, in coordinating inflammation and host defense. Toll-like receptor (TLR) involvement in early activation events within the mesothelium, however, remains poorly defined. To investigate the initiation of bacterial peritonitis, we characterized TLR activation by bacterial ligands in human peritoneal mesothelial cells (HPMC). METHODS: Primary HPMC were isolated from omental biopsies and TLR expression detected by real-time polymerase chain reaction (PCR), reverse transcription (RT) PCR and flow cytometry. The responsiveness of HPMC to specific bacterial TLR agonists was determined using chemokine production as a biological readout. The requirement for CD14 in HPMC responses to a clinically relevant Staphylococcus epidermidis cell-free supernatant (SES) was investigated using soluble CD14 or anti-CD14-blocking antibodies. RESULTS: Real-time PCR detected TLR1-6 messenger RNA expression in HPMC and responses to TLR2/1 and TLR2/6 ligands and SES. No cell surface TLR4 expression or responses to lipopolysaccharide were detectable in HPMC, but they did respond to flagellin, a TLR5 ligand. SES-mediated responses were dependent on TLR2 but did not require CD14 in HPMC for optimal efficiency, unlike peripheral blood mononuclear cells. HPMC expression of TLR2 was also modulated by TLR2 ligands and inflammatory cytokines. CONCLUSIONS: These data suggest that mesothelial cell activation by TLR2/1, TLR2/6 and TLR5 contributes to bacterial recognition influencing the course of the infective process and has implications for improving treatment of infection in PD patients. PMID- 21633097 TI - An immunofluorescence test for phospholipase-A2-receptor antibodies and its clinical usefulness in patients with membranous glomerulonephritis. AB - BACKGROUND: The recent finding that phospholipase-A(2)-receptor antibodies (PLA(2)R-AB) may play a role in the development of primary membranous glomerulonephritis (MGN) offers the opportunity to measure a marker to help diagnose, classify and eventually monitor the course of patients with MGN. METHODS: We developed an immunofluorescence test, which allows the easy and specific analysis of the presence of PLA(2)R-AB in serum. The usefulness of this test was studied in 153 healthy blood donors, 90 patients with non-membranous glomerular injuries, 17 patients with a secondary form of MGN and 100 patients with biopsy-proven primary MGN. In addition, in five patients with biopsy-proven MGN, PLA(2)R-AB levels were monitored prospectively for up to 18 months following a single dose of rituximab (RTX) (375 mg/m(2) body surface). RESULTS: PLA(2)R-AB were not found in healthy controls or patients with glomerular lesions other than biopsy-proven primary MGN. Fifty-two patients with primary MGN (52%) were positive for PLA(2)R-AB. The levels ranged from 1:10 to 1:3200. In patients who had MGN and were treated with RTX the fall in PLA(2)R-AB levels was followed by a decrease in proteinuria, whereas an increase in PLA(2)R-AB levels was associated with an increase in proteinuria. CONCLUSIONS: These studies show that the new test allows the monitoring of PLA(2)R-AB levels in patients with MGN and may help in making therapeutic decisions for these patients. PMID- 21633098 TI - Recent developments in cardiovascular risk assessment: relevance to the nephrologist. PMID- 21633099 TI - Malignancies after renal transplantation in Taiwan: a nationwide population-based study. AB - BACKGROUND: Renal transplantation has been regarded as the treatment of choice for end-stage renal disease. Renal transplantation increases the risk of cancers due to long-term immunosuppression. The types of post-transplantation malignancies may vary among different geographic regions and ethnic populations. To date, large population-based studies of post-transplantation malignancies in Asian renal transplant recipients (RTRs) have rarely been reported. METHODS: To investigate the patterns of post-transplantation malignancies in Chinese RTRs, we performed a nationwide population-based cohort study between 1997 and 2008 based on data from the National Health Insurance Database in Taiwan. Patterns of cancer incidence in RTRs were compared with those of the general population using standardized incidence ratios (SIRs). RESULTS: Among the 4716 RTRs (2475 males and 2241 females; mean age 44.1 +/- 12.4 years) and 22 556 person-years of observation, 320 post-transplant cancers were diagnosed. The SIR of all cancers was 3.75 (95% confidence interval 3.36-4.18). Women had a higher risk than men for the development of malignancies (SIR 5.04 for women and SIR 2.88 for men). Renal, bladder and liver cancers were the most common cancers, with SIRs of 44.29, 42.89 and 5.07, respectively. When stratified by age, RTRs of young age at transplant (<20 years) had the highest risk of post-transplantation malignancies. CONCLUSIONS: This study demonstrates different patterns of malignancies after renal transplantation in Chinese RTRs, with higher incidences of kidney and bladder cancers. Physicians should be more vigilant in examining RTRs for post transplantation malignancies especially in younger patients. PMID- 21633100 TI - Angiotensin II receptor blocker pretreatment of rats undergoing sudden renal ablation. AB - BACKGROUND: Subtotal nephrectomy (N) in rats results in progressive hypertension, proteinuria and renal lesions. Renin-angiotensin system blockade initiated at N prevents these changes; treatments failing to reduce hypertension and proteinuria do not. METHODS: Ten Munich-Wistar rats underwent 1½ surgical N; eight littermates were pretreated with losartan (L) only for 6 weeks prior to 1½ N (N + L). Pretreated (n = 8; C + L) and untreated controls (C; n = 8) had sham operations. RESULTS: Over 6 months, N and N + L rats developed ~80% increase in glomerular filtration rate per nephron over C and C + L, P < 0.001). Hypertension (intra-arterial mean blood pressure 116 +/- 6.8 mmHg in N rats versus 102 +/- 3.2 in C, 104 +/- 8.4 in C + L, and 104 +/- 8.4 in N + L rats, P < 0.001 for all) and proteinuria (120 +/- 20 mg/day in N versus 39 +/- 10 in C, 34 +/- 8 in C + L and 35 +/- 8 in N + L, P < 0.001 for all) developed only in N. Focal segmental glomerulosclerosis (FSGS) (%) at 6 months was 20 +/- 8 in N and 17.5 +/- 8 in N + L (ns) and <1 in C and C + L (P < 0.001 versus N and N + L). Interstitial fractional volume (Vv), 4.0 +/- 1.7% in C and 4.4 +/- 1.6% in C + L (ns), was similarly increased to 7.5 +/- 2.5% in N and 9.0 +/- 3.9% in N+L (P < 0.04 versus C and C + L). Atrophic tubule Vv was increased by >300% in N and N + L over C and C + L (P < 0.02 for all). Glomerular volume doubled in N and N + L (P < 0.001). Podocyte foot process effacement was greater in N and NL than in C or C + L (P <= 0.02 for all). Thus, L given for 6 weeks prior to 1½ N prevented hypertension and proteinuria over the subsequent 6 months without reducing glomerular hypertrophy, hyperfiltration or interstitial, tubular or FSGS lesions or foot process effacement. CONCLUSIONS: These studies dissociated systemic hypertension and proteinuria from the renal lesions in this model. Durable effects of losartan on blood pressure and proteinuria likely represent epigenetic processes. PMID- 21633101 TI - Use of aminoglycosides for peritoneal dialysis-associated peritonitis does not affect residual renal function. AB - BACKGROUND: Aminoglycosides offer several potential benefits in their treatment of peritoneal dialysis (PD)-associated peritonitis, including low cost, activity against Gram-negative organisms (including Pseudomonas aeruginosa), synergistic bactericidal activity against some Gram-positive organisms (such as Staphylococci) and relatively low propensity to promote antimicrobial resistance. However, there is limited conflicting evidence that aminoglycosides may accelerate loss of residual renal function (RRF) in PD patients. The aim of this study was to study the effect of aminoglycoside use on slope of decline in RRF. METHODS: The study included 2715 Australian patients receiving PD between October 2003 and December 2007 in whom at least two measurements of renal creatinine clearance were available. Patients were divided according to tertiles of slope of RRF decline (rapid, intermediate and slow). The primary outcome was the slope of RRF over time in patients who received aminoglycosides for PD peritonitis versus those who did not. RESULTS: A total of 1412 patients (52%) experienced at least one episode of PD peritonitis. An aminoglycoside was used as the initial empiric antibiotic in 1075 patients. The slopes of RRF decline were similar in patients treated and not treated with at least one course of aminoglycoside (median [interquartile range] -0.26 [-1.17 to 0.04] mL/min/1.73 m(2)/month versus -0.22 [ 1.11 to 0.01] mL/min/1.73 m(2)/month, P = 0.9). The slopes of RRF decline were also similar in patients receiving repeated courses of aminoglycoside. CONCLUSIONS: Empiric treatment with aminoglycoside for peritonitis was not associated with an adverse effect on RRF in PD patients. PMID- 21633102 TI - Proliferative glomerulonephritis with monoclonal IgG deposits secondary to chronic lymphocytic leukemia. Report of two cases. AB - Proliferative glomerulonephritis with monoclonal IgG deposits (PGNMID) is a recently described entity that is only rarely associated with a hematological or lymphoproliferative malignancy. We describe the cases of two men with preexisting chronic lymphocytic leukemia (CLL) who developed endocapillary proliferative glomerulonephritis with nonorganized monoclonal IgG(1) deposits. One biopsy also showed CLL infiltration of the cortex. Both patients were treated with rituximab in addition to cyclophosphamide in one case and fludarabine in the other with significant improvement of their renal disease and CLL. This report provides additional evidence to support the use of rituximab in the therapy of CLL associated PGNMID. PMID- 21633103 TI - Endovascular treatment of a pial arteriovenous fistula with occipital remodeling secondary to giant torcular dilation. AB - Intracranial arteriovenous fistulas are vascular malformations in which clinical suspicion and prompt diagnosis, with a subsequent appropriate therapeutic approach, are crucial to avoid the development of irreversible neurological damage or even patient death because these lesions can be associated with heavy bleeding and high mortality rates. The authors present the case of a direct pial arteriovenous fistula in an infant and its unusual presentation as a large occipital protuberance with hard consistency and an audible murmur that produced bone remodeling. The patient was successfully treated by endovascular therapy, with complete regression of the occipital protuberance during follow-up. PMID- 21633105 TI - Comparison of corrected QT interval as measured on electroencephalography versus 12-lead electrocardiography in children with a history of syncope. AB - Long QT syndrome can present with neurological manifestations, including syncope and seizure-like activity. These patients often receive an initial neurologic evaluation, including electroencephalography (EEG). Our previous retrospective study suggested an increased prevalence of prolonged corrected QT interval (QTc) measured during the EEG of patients with syncope. The aim of the current study is to assess the accuracy of the EEG QTc reading compared with the nonsimultaneous 12-lead electrocardiography (ECG) in children with syncope. Abnormal QTc was defined as >=450 ms in boys, >=460 ms in girls. Forty-two children were included. There was no significant correlation between QTc readings in the EEG and ECG. EEG failed to identify 2 children with prolonged QTc in the ECG and overestimated the QTc in 3 children with normal QTc in the ECG. This study suggests that interpretation of the QTc segment during an EEG is limited. Further studies with simultaneous EEG and 12-lead ECG are warranted. PMID- 21633104 TI - Childhood hemorrhagic stroke: an important but understudied problem. AB - Hemorrhagic stroke in children occurs more frequently than is commonly appreciated. There are important differences in the factors associated with hemorrhagic stroke in children when compared with adults. These differences likely play a role in the different outcomes, which tend to worsen with age. In this review, we describe the estimated frequency, clinical presentation, acute management of hemorrhagic stroke, and an overview of rehabilitation techniques. We identify key topics for future basic and clinical research. Findings from future studies will help improve our ability to optimize treatment for and long term rehabilitation of these patients. PMID- 21633106 TI - Magnetic resonance imaging findings in Reye syndrome: case report and review of the literature. AB - Magnetic resonance imaging findings in Reye syndrome have been reported only infrequently. A previously well 8-year-old boy presented with repeated episodes of vomiting and abdominal pain followed by altered sensorium and tonic spasms. This occurred 5 days after upper respiratory tract infection. His laboratory data revealed elevated liver enzymes, prolonged prothrombin time, and high blood ammonia levels. Magnetic resonance imaging of the brain done on the day of admission revealed diffuse cerebral edema and signal alterations in brainstem, bilateral thalami, medial temporal lobes, parasagittal cortex, and cerebellar and subcortical white matter. Diffusion restriction was seen in thalami, midbrain, cerebellar white matter, subcortical white matter, and parasaggital cortex in the watershed territory. The patient made a full recovery. Follow-up magnetic resonance imaging after a week revealed complete resolution of all except thalamic lesions. Although diffusion restriction in thalami and midbrain has been reported previously, this is the first report indicating diffusion restriction in subcortical white matter and the parasagittal cortex. PMID- 21633107 TI - Functional interaction between dynein light chain and intermediate chain is required for mitotic spindle positioning. AB - Cytoplasmic dynein is a large multisubunit complex involved in retrograde transport and the positioning of various organelles. Dynein light chain (LC) subunits are conserved across species; however, the molecular contribution of LCs to dynein function remains controversial. One model suggests that LCs act as cargo-binding scaffolds. Alternatively, LCs are proposed to stabilize the intermediate chains (ICs) of the dynein complex. To examine the role of LCs in dynein function, we used Saccharomyces cerevisiae, in which the sole function of dynein is to position the spindle during mitosis. We report that the LC8 homologue, Dyn2, localizes with the dynein complex at microtubule ends and interacts directly with the yeast IC, Pac11. We identify two Dyn2-binding sites in Pac11 that exert differential effects on Dyn2-binding and dynein function. Mutations disrupting Dyn2 elicit a partial loss-of-dynein phenotype and impair the recruitment of the dynein activator complex, dynactin. Together these results indicate that the dynein-based function of Dyn2 is via its interaction with the dynein IC and that this interaction is important for the interaction of dynein and dynactin. In addition, these data provide the first direct evidence that LC occupancy in the dynein motor complex is important for function. PMID- 21633108 TI - Overlapping kinetochore targets of CK2 and Aurora B kinases in mitotic regulation. AB - Protein kinase CK2 is one of the most conserved kinases in eukaryotic cells and plays essential roles in diverse processes. While we know that CK2 plays a role(s) in cell division, our understanding of how CK2 regulates cell cycle progression is limited. In this study, we revealed a regulatory role for CK2 in kinetochore function. The kinetochore is a multi-protein complex that assembles on the centromere of a chromosome and functions to attach chromosomes to spindle microtubules. To faithfully segregate chromosomes and maintain genomic integrity, the kinetochore is tightly regulated by multiple mechanisms, including phosphorylation by Aurora B kinase. We found that a loss of CK2 kinase activity inhibits anaphase spindle elongation and results in chromosome missegregation. Moreover, a lack of CK2 activates the spindle assembly checkpoint. We demonstrate that CK2 associates with Mif2, the Saccharomyces cerevisiae homologue of human CENP-C, which serves as an important link between the inner and outer kinetochore. Furthermore, we show Mif2 and the inner kinetochore protein Ndc10 are phosphorylated by CK2, and this phosphorylation plays antagonistic and synergistic roles with Aurora B phosphorylation of these targets, respectively. PMID- 21633109 TI - The Caenorhabditis elegans paxillin orthologue, PXL-1, is required for pharyngeal muscle contraction and for viability. AB - We have identified the gene C28H8.6 (pxl-1) as the Caenorhabditis elegans orthologue of vertebrate paxillin. PXL-1 contains the four C-terminal LIM domains conserved in paxillin across all species and three of the five LD motifs found in the N-terminal half of most paxillins. In body wall muscle, PXL-1 antibodies and a full-length green fluorescent protein translational fusion localize to adhesion sites in the sarcomere, the functional repeat unit in muscle responsible for contraction. PXL-1 also localizes to ring-shaped structures near the sarcolemma in pharyngeal muscle corresponding to podosome-like sites of actin attachment. Our analysis of a loss-of-function allele of pxl-1, ok1483, shows that loss of paxillin leads to early larval arrested animals with paralyzed pharyngeal muscles and eventual lethality, presumably due to an inability to feed. We rescued the mutant phenotype by expressing paxillin solely in the pharynx and found that these animals survived and are essentially wild type in movement and body wall muscle structure. This indicates a differential requirement for paxillin in these two types of muscle. In pharyngeal muscle it is essential for contraction, whereas in body wall muscle it is dispensable for filament assembly, sarcomere stability, and ultimately movement. PMID- 21633110 TI - A mechanism of Rap1-induced stabilization of endothelial cell--cell junctions. AB - Activation of Rap1 small GTPases stabilizes cell--cell junctions, and this activity requires Krev Interaction Trapped gene 1 (KRIT1). Loss of KRIT1 disrupts cardiovascular development and causes autosomal dominant familial cerebral cavernous malformations. Here we report that native KRIT1 protein binds the effector loop of Rap1A but not H-Ras in a GTP-dependent manner, establishing that it is an authentic Rap1-specific effector. By modeling the KRIT1-Rap1 interface we designed a well-folded KRIT1 mutant that exhibited a ~40-fold-reduced affinity for Rap1A and maintained other KRIT1-binding functions. Direct binding of KRIT1 to Rap1 stabilized endothelial cell-cell junctions in vitro and was required for cardiovascular development in vivo. Mechanistically, Rap1 binding released KRIT1 from microtubules, enabling it to locate to cell--cell junctions, where it suppressed Rho kinase signaling and stabilized the junctions. These studies establish that the direct physical interaction of Rap1 with KRIT1 enables the translocation of microtubule-sequestered KRIT1 to junctions, thereby supporting junctional integrity and cardiovascular development. PMID- 21633111 TI - Topological arrangement of the intracellular membrane fusion machinery. AB - Soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) form a four-helix coiled-coil bundle that juxtaposes two bilayers and drives a basal level of membrane fusion. The Sec1/Munc18 (SM) protein binds to its cognate SNARE bundle and accelerates the basal fusion reaction. The question of how the topological arrangement of the SNARE helices affects the reactivity of the fusion proteins remains unanswered. Here we address the problem for the first time in a reconstituted system containing both SNAREs and SM proteins. We find that to be fusogenic a SNARE topology must support both basal fusion and SM stimulation. Certain topological combinations of exocytic SNAREs result in basal fusion but cannot support SM stimulation, whereas other topologies support SM stimulation without inducing basal fusion. It is striking that of all the possible topological combinations of exocytic SNARE helices, only one induces efficient fusion. Our results suggest that the intracellular membrane fusion complex is designed to fuse bilayers according to one genetically programmed topology. PMID- 21633112 TI - Accelerated and adaptive evolution of yeast sexual adhesins. AB - There is a recent emergence of interest in the genes involved in gametic recognition as drivers of reproductive isolation. The recent population genomic sequencing of two species of sexually primitive yeasts (Liti G, Carter DM, Moses AM, Warringer J, Parts L, James SA, Davey RP, Roberts IN, Burt A, Koufopanou V et al. [23 co-authors]. 2009. Population genomics of domestic and wild yeasts. Nature 458:337-341.) has provided data for systematic study of the roles these genes play in the early evolution of sex and speciation. Here, we discovered that among genes encoding cell surface proteins, the sexual adhesin genes have evolved significantly more rapidly than others, both within and between Saccharomyces cerevisiae and its closest relative S. paradoxus. This result was supported by analyses using the PAML pairwise model, a modified McDonald-Kreitman test, and the PAML branch model. Moreover, using a combination of a new statistic of neutrality, an information theory-based measure of evolutionary variability, and functional characterization of amino acid changes, we found that a higher proportion of amino acid changes are fixed in the sexual adhesins than in other proteins and a greater proportion of the fixed amino acid changes either between the two species or the two subgroups of S. paradoxus are functionally dissimilar or radically different. These results suggest that the accelerated evolution of sexual adhesin genes may facilitate speciation, or incipient speciation, and promote sexual selection in general. PMID- 21633113 TI - Retroposon insertions and the chronology of avian sex chromosome evolution. AB - The vast majority of extant birds possess highly differentiated Z and W sex chromosomes. Nucleotide sequence data from gametologs (homologs on opposite sex chromosomes) suggest that this divergence occurred throughout early bird evolution via stepwise cessation of recombination between identical sex chromosomal regions. Here, we investigated avian sex chromosome differentiation from a novel perspective, using retroposon insertions and random insertions/deletions for the reconstruction of gametologous gene trees. Our data confirm that the CHD1Z/CHD1W genes differentiated in the ancestor of the neognaths, whereas the NIPBLZ/NIPBLW genes diverged in the neoavian ancestor and independently within Galloanserae. The divergence of the ATP5A1Z/ATP5A1W genes in galloanserans occurred independently in the chicken, the screamer, and the ancestor of duck-related birds. In Neoaves, this gene pair differentiated in each of the six sampled representatives, respectively. Additionally, three of our investigated loci can be utilized as universal, easy-to-use independent tools for molecular sexing of Neoaves or Neognathae. PMID- 21633114 TI - Reconciling gene and genome duplication events: using multiple nuclear gene families to infer the phylogeny of the aquatic plant family Pontederiaceae. AB - Most plant phylogenetic inference has used DNA sequence data from the plastid genome. This genome represents a single genealogical sample with no recombination among genes, potentially limiting the resolution of evolutionary relationships in some contexts. In contrast, nuclear DNA is inherently more difficult to employ for phylogeny reconstruction because major mutational events in the genome, including polyploidization, gene duplication, and gene extinction can result in homologous gene copies that are difficult to identify as orthologs or paralogs. Gene tree parsimony (GTP) can be used to infer the rooted species tree by fitting gene genealogies to species trees while simultaneously minimizing the estimated number of duplications needed to reconcile conflicts among them. Here, we use GTP for five nuclear gene families and a previously published plastid data set to reconstruct the phylogenetic backbone of the aquatic plant family Pontederiaceae. Plastid-based phylogenetic studies strongly supported extensive paraphyly of Eichhornia (one of the four major genera) but also depicted considerable ambiguity concerning the true root placement for the family. Our results indicate that species trees inferred from the nuclear genes (alone and in combination with the plastid data) are highly congruent with gene trees inferred from plastid data alone. Consideration of optimal and suboptimal gene tree reconciliations place the root of the family at (or near) a branch leading to the rare and locally restricted E. meyeri. We also explore methods to incorporate uncertainty in individual gene trees during reconciliation by considering their individual bootstrap profiles and relate inferred excesses of gene duplication events on individual branches to whole-genome duplication events inferred for the same branches. Our study improves understanding of the phylogenetic history of Pontederiaceae and also demonstrates the utility of GTP for phylogenetic analysis. PMID- 21633115 TI - Dose-response assessment of fetal testosterone production and gene expression levels in rat testes following in utero exposure to diethylhexyl phthalate, diisobutyl phthalate, diisoheptyl phthalate, and diisononyl phthalate. AB - Several phthalate esters have been linked to the Phthalate Syndrome, affecting male reproductive development when administered to pregnant rats during in utero sexual differentiation. The goal of the current study was to enhance understanding of this class of compounds in the Sprague Dawley (SD) fetal rat following exposure on gestational days (GDs) 14-18 by determining the relative potency factors for several phthalates on fetal testes endpoints, the effects of a nine phthalate mixture on fetal testosterone (T) production, and differences in SD and Wistar (W) strain responses of fetal T production and testicular gene expression to di(2-ethylhexyl) phthalate (DEHP). We determined that diisobutyl phthalate (DIBP) and diisoheptyl phthalate (DIHP) reduced fetal testicular T production with similar potency to DEHP, whereas diisononyl phthalate (DINP) was 2.3-fold less potent. DINP was also less potent at reducing StAR and Cyp11a gene expression levels, whereas DIBP was slightly more potent than DEHP. We observed that administration of dilutions of a mixture of nine phthalates (DEHP, DIHP, DIBP, dibutyl-, benzyl butyl-, dicyclohexyl-, diheptyl-, dihexyl-, and dipentyl phthalate) reduced fetal T production in a dose-dependent manner best predicted by dose addition. Finally, we found that the differential effects of in utero DEHP treatment on epididymal and gubernacular differentiation in male SD and W rats (0, 100, 300, 500, 625, 750, or 875 mg DEHP/kg/day) are likely due to tissue specific strain differences in the androgen and insl3 signaling pathways rather than differential effects of DEHP on fetal testis T and insl3 production. PMID- 21633116 TI - Nicotine exposure during the third trimester equivalent of human gestation: time course of effects on the central cholinergic system of rats. AB - Up to 22% of pregnant women smoke, which constitutes a major health concern. Nicotine, a cholinergic agonist, causes deleterious effects on brain development. However, most studies investigate its effects during rodents' gestation, which corresponds, in terms of neural development, to the first two trimesters of human gestation. Here, we focused on effects of nicotine on the brain cholinergic system during the third trimester equivalent of human gestation. From the 2nd to the 19th day of lactation, dams were exposed either to nicotine (6 mg/kg/day) or to saline via sc osmotic minipumps. Offspring were sacrificed during exposure (PN15, PN, postnatal) or at 2 days (PN21), 11 days (PN30), or 10 weeks (PN90) of withdrawal. In the cerebral cortex, midbrain, and hippocampus, we assessed nicotinic acetylcholine receptor (nAChR) binding, [(3)H]hemicholinium-3 (HC-3) binding to the high-affinity choline transporter, choline acetyltransferase (ChAT), and acetylcholinesterase (AChE) activities. Nicotine-exposed offspring presented nAChR upregulation during exposure in all brain regions, reduced HC-3 binding during and 11 days postexposure, and increased HC-3 binding on PN90. Effects on ChAT and AChE were dependent on the brain region and restricted to the withdrawal period: There were increased activities in the midbrain on PN30. In the hippocampus, AChE as reduced on PN30, whereas, for ChAT, the decrease was followed by late-emergent increased activity. These data indicate that maternal nicotine exposure during the third trimester equivalent of human gestation promotes cholinergic system alterations in the offspring's brain. In addition, detrimental effects are observable even long after the exposure has been interrupted. PMID- 21633117 TI - Detection of parent-of-origin effects for quantitative traits in complete and incomplete nuclear families with multiple children. AB - For a diallelic genetic marker locus, tests like the parental-asymmetry test (PAT) are simple and powerful for detecting parent-of-origin effects. However, these approaches are applicable only to qualitative traits and thus are currently not suitable for quantitative traits. In this paper, the authors propose a novel class of PAT-type parent-of-origin effects tests for quantitative traits in families with both parents and an arbitrary number of children, which is denoted by Q-PAT(c) for some constant c. The authors further develop Q-1-PAT(c) for detection of parent-of-origin effects when information is available on only 1 parent in each family. The authors suggest the Q-C-PAT(c) test for combining families with data on both parental genotypes and families with data on only 1 parental genotype. Simulation studies show that the proposed tests control the empirical type I error rates well under the null hypothesis of no parent-of origin effects. Power comparison also demonstrates that the proposed methods are more powerful than the existing likelihood ratio test. Although normality is commonly assumed in methods for studying quantitative traits, the tests proposed in this paper do not make any assumption about the distribution of the quantitative trait. PMID- 21633118 TI - Should gestational weight gain recommendations be tailored by maternal characteristics? AB - The authors tested whether the relation between gestational weight gain (GWG) and 5 adverse pregnancy outcomes (small-for-gestational-age (SGA) birth, large-for gestational-age (LGA) birth, spontaneous preterm birth, indicated preterm birth, and unplanned cesarean delivery) differed according to maternal race/ethnicity, smoking, parity, age, and/or height. They also evaluated whether GWG guidelines should be modified for special populations by studying GWG and risk of at least 1 adverse outcome within different subgroups. Data came from a cohort of 23,362 normal-weight mothers who delivered singletons at Magee-Womens Hospital in Pittsburgh, Pennsylvania (2003-2008). Adequacy of GWG was defined as observed GWG divided by recommended GWG. The synergy analysis found that the combination of smoking, black race/ethnicity, primiparity, or short height with poor GWG was associated with an excess risk of SGA birth, while high GWG combined with each of these characteristics diminished risk of LGA birth in comparison with the same GWG among the women's counterparts. Nevertheless, there were no significant or meaningful differences in the risk of at least 1 adverse outcome between the GWG recommended by the Institute of Medicine in 2009 and the GWG that minimized risk of the composite outcome. These findings do not support the tailoring of GWG guidelines on the basis of a mother's smoking status, race/ethnicity, parity, age, or height among normal-weight women. PMID- 21633119 TI - Physical exercise, body mass index, and risk of chronic pain in the low back and neck/shoulders: longitudinal data from the Nord-Trondelag Health Study. AB - Chronic musculoskeletal pain constitutes a large socioeconomic challenge, and preventive measures with documented effects are warranted. The authors' aim in this study was to prospectively investigate the association between physical exercise, body mass index (BMI), and risk of chronic pain in the low back and neck/shoulders. The study comprised data on approximately 30,000 women and men in the Nord-Trondelag Health Study (Norway) who reported no pain or physical impairment at baseline in 1984-1986. Occurrence of chronic musculoskeletal pain was assessed at follow-up in 1995-1997. A generalized linear model was used to calculate adjusted risk ratios. For both females and males, hours of physical exercise per week were linearly and inversely associated with risk of chronic pain in the low back (women: P-trend = 0.02; men: P-trend < 0.001) and neck/shoulders (women: P-trend = 0.002; men: P-trend < 0.001). Obese women and men had an approximately 20% increased risk of chronic pain in both the low back and the neck/shoulders. Exercising for 1 or more hours per week compensated, to some extent, for the adverse effect of high BMI on risk of chronic pain. The authors conclude that physical inactivity and high BMI are associated with an increased risk of chronic pain in the low back and neck/shoulders in the general adult population. PMID- 21633120 TI - Inclusion of African Americans in genetic studies: what is the barrier? AB - To facilitate an increase in the amount of data on minority subjects collected for genetic databases, the authors attempted to clarify barriers to African American participation in genetic studies. They randomly sampled 78,072 subjects from the community (Missouri Family Registry, 2002-2007). Of these, 28,658 participated in a telephone screening interview, 3,179 were eligible to participate in the genetic study, and 1,919 participated in the genetic study. Response rates were examined in relation to the proportion of subjects in the area who were African-American according to US Census 2000 zip code demographic data. Compared with zip codes with fewer than 5% African Americans (average = 2% African-American), zip codes with at least 60% African Americans (average = 87% African-American) had higher proportions of subjects with an incorrect address or telephone number but lower proportions of subjects who did not answer the telephone and subjects who refused the telephone interview (P < 0.0001). Based on reported race from the telephone screening, 71% of eligible African Americans and 57% of eligible European Americans participated in the genetic study (P < 0.0001). The results of this study suggest that increasing the number of African Americans in genetic databases may be achieved by increasing efforts to locate and contact them. PMID- 21633122 TI - Long-term perimetric fluctuation in glaucoma. PMID- 21633121 TI - C-reactive protein and complement factor H in aged human eyes and eyes with age related macular degeneration. AB - BACKGROUND: There is increasing evidence that inflammation and immune-mediated processes (complement activation) play an important role in age-related macular degeneration (AMD) pathogenesis. A genetic variation in the gene encoding complement factor H (CFH) and plasma levels of C-reactive protein (CRP), a systemic marker of subclinical inflammation, have consistently been shown to be associated with an increased risk for AMD. In the present study, we examined the immunolocalisation of CRP and CFH in aged control human donor eyes (n=10; mean age 79 years) and eyes with AMD (n=18; mean age 83 years). METHODS: Alkaline phosphatase immunohistochemistry was performed using polyclonal antibodies against CRP and CFH on cryopreserved tissue sections from disc/macular blocks. Three independent masked observers scored the reaction product (0-8). RESULTS: In aged control eyes, the retinal pigment epithelium/Bruch's membrane/choriocapillaris (RPE/BrM/CC) complex including intercapillary septa (ICS) had the most prominent immunostaining for CRP and CFH. CRP was significantly higher than controls in BrM/CC/ICS and choroidal stroma in early and wet AMD eyes (p<0.05). In contrast, CFH was significantly lower in BrM/CC/ICS complex of AMD choroids than in controls (p<0.05). Interestingly, CRP and CFH were significantly reduced in BrM/CC/ICS complex in atrophic area of macula in geographical atrophy (p<0.05). Drusen and basal laminar deposits were intensely positive for CRP and CFH. CONCLUSION: These immunohistochemical findings show that changes in distribution and relative levels of CRP and CFH were evident in early and late AMD eyes. This suggests that high levels of CRP and insufficient CFH at the retina/choroid interface may lead to uncontrolled complement activation with associated cell and tissue damage. This study supports the hypothesis that inflammation and immune-mediated mechanisms are involved in the pathogenesis of AMD. PMID- 21633123 TI - Increasing depressive symptoms in children of Alzheimer parents. AB - Children of persons with Alzheimer disease (AD) are at increased risk of developing AD themselves, but specific factors that predict AD in this population have yet to be elucidated. Various studies indicate depressive symptoms may predate clinical AD and represent a risk factor and/or prodrome of incipient dementia, but their relevance for AD offspring remains uncertain. As part of a longitudinal family study of AD, we assessed depressive symptomatology in 30 middle aged AD offspring (mean age at baseline: 41.2). Their mean total scores on the Hamilton Depression Rating scale increased from 1.8 to 5.3 (P < .001) across a 20-year interval. Neurocognitive performance remained stable in a subset of this cohort (N = 25) over the same interval. Findings from this small convenience sample suggest emerging depressive symptoms may be among the earliest signs of subsequent dementia in this high-risk population but require confirmation through further longitudinal follow-up and replication in larger populations. PMID- 21633124 TI - Midlife motivational abilities predict apathy and depression in Alzheimer disease: the aging, demographics, and memory study. AB - Apathy and depression are the most common neuropsychiatric symptoms in mild cognitive impairment (MCI) and Alzheimer disease (AD). This study was the first to explore midlife motivational abilities as a predictor of the progression of apathy and depression in MCI and AD. It used a subsample of the Aging, Demographics, and Memory Study (N = 137). Participants, aged over 70, were categorized according to baseline clinical diagnosis (normal cognition, MCI, or AD). Assessments were conducted at an 18-month interval. Neuropsychiatric symptoms were assessed using the Neuropsychiatric Inventory. Midlife motivational abilities were estimated on the basis of the main occupation using the Occupational Information Network (O*NET) database, which provides detailed information on worker abilities. Repeated measures analysis of covariance was used. Apathy and depression were found to be particularly high in participants with AD and high motivational abilities. Apathy, but not depression, increased over time in those with AD and high motivational abilities. It would appear that holding on to unattainable goals with strong motivational efforts when faced with severe cognitive loss might lead to unproductive persistence, depressive reaction, and more apathetic behavior. PMID- 21633125 TI - A three-stage-integrative approach for the identification of potential hepatotoxic compounds from botanical products. AB - With the increasing use of herbal medicines and dietary supplements, intensive concerns about their potential toxicities have been raised. Screening and identifying the toxic compounds from these botanical products composed by hundreds of components have become a critical but challenging problem. In this study, 3 methods, including fraction separation, an in-house-developed fluorescein diacetate-based automatic microscopy screening (FAMS) platform, and liquid chromatography-mass spectrometry-based compounds identification were integrated within the Three-Stage-Integrative (TSI) approach for the identification of potential hepatotoxicants from botanical products. The sensitivity and linear range of FAMS assay was validated and compared with 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay by previously reported hepatotoxic compounds. The success of TSI approach was further demonstrated by its application to Fructus aristolochiae. Aristolochic acid IVa and aristolodione were tentatively identified to be potential hepatotoxicants in this plant. These applications suggested that our TSI approach provides an effective tool for identifying potential toxic compounds from botanical products. PMID- 21633126 TI - Toxicity profile of solvents by aspiration approach for topical agent delivery to respiratory tract epithelium. AB - Agent solubility is a problem for aspiration of agents into lungs for chemopreventive efficacy evaluation, since many agents have to be dissolved in solvents. These solvents may be toxic to the lung epithelium. A study was conducted in female A/J mice to determine toxicity of different solvent concentrations by using saline, dimethyl sulfoxide (DMSO), ethanol, polyethylene glycol 400 (PEG-400), and labrasol for 1, 5, and 28 days via aspiration route. Toxicity was determined by measuring changes in body weight and bronchoalveolar lavage fluid (BALF). No significant difference was observed in body weight, differential cell counts, lactate dehydrogenase (LDH) and total protein in all solvent groups compared to saline by 28 days except 50% ethanol. However, myeloperoxidase (MPO) and macrophage inflammatory protein-2 (MIP-2) showed significant increase in 2% and 10% DMSO, 10% ethanol, 0.1% and 2% PEG-400, and 1% labrasol by longer dosing. All solvents except for 10% ethanol and 2% PEG-400 are suitable for agent aspiration. PMID- 21633127 TI - Effect of enoxacin, felbinac, and sparfloxacin on fatty acid metabolism and glucose concentrations in rat tissues. AB - Multiple changes in metabolic levels could be useful for understanding physiological toxicity. To explore further risk factors for the convulsions induced by the interaction of nonsteroidal anti-inflammatory and new quinolone antimicrobial drugs, the effect of sparfloxacin, enoxacin, and felbinac on fatty acid metabolism and glucose concentrations in the liver, brain, and blood of rats was investigated. The levels of long-chain acyl-CoAs (C(18:1) and C(20:4)) in the liver and brain were decreased at the onset of convulsions induced by the coadministration of enoxacin with felbinac. Then, glucose concentrations in the liver and blood were decreased, whereas they were increased in a dose-dependant manner in the brain. However, the formation of acyl-CoAs and glucose levels in the liver, brain, and blood was not significantly influenced by enoxacin, felbinac, and sparfloxacin alone, respectively. The disturbance of both fatty acid metabolism and glucose levels might be associated with the increased susceptibility to convulsions, which may contribute to further understanding of the toxic effects associated with these drugs. PMID- 21633128 TI - Quantitative EEG as a predictive biomarker for Parkinson disease dementia. AB - OBJECTIVE: We evaluated quantitative EEG (QEEG) measures as predictive biomarkers for the development of dementia in Parkinson disease (PD). Preliminary work shows that QEEG measures correlate with current PD cognitive state. A reliable predictive QEEG biomarker for PD dementia (PD-D) incidence would be valuable for studying PD-D, including treatment trials aimed at preventing cognitive decline in PD. METHODS: A cohort of subjects with PD in our brain donation program utilizes annual premortem longitudinal movement and cognitive evaluation. These subjects also undergo biennial EEG recording. EEG from subjects with PD without dementia with follow-up cognitive evaluation was analyzed for QEEG measures of background rhythm frequency and relative power in delta, , alpha, and beta bands. The relationship between the time to onset of dementia and QEEG and other possible predictors was assessed by using Cox regression. RESULTS: The hazard of developing dementia was 13 times higher for those with low background rhythm frequency (lower than the grand median of 8.5 Hz) than for those with high background rhythm frequency (p < 0.001). Hazard ratios (HRs) were also significant for > median bandpower (HR = 3.0; p = 0.004) compared to below, and for certain neuropsychological measures. The HRs for delta, alpha, and beta bandpower as well as baseline demographic and clinical characteristics were not significant. CONCLUSION: The QEEG measures of background rhythm frequency and relative power in the band are potential predictive biomarkers for dementia incidence in PD. These QEEG biomarkers may be useful in complementing neuropsychological testing for studying PD-D incidence. PMID- 21633129 TI - Seize the day: Quantitative EEG as a biomarker for dementia in Parkinson disease. PMID- 21633130 TI - Local response following the Great East Japan Earthquake 2011. PMID- 21633131 TI - The influence of physical activity on cardiometabolic biomarkers in youths: a review. AB - The purposes of this article were to (1): review recent studies of relations between physical activity and cardiometabolic biomarkers of youths (2); highlight areas in which additional research is needed; and (3) make recommendations for preventive interventions. Observational studies show that youths who engage in high amounts of moderate-vigorous physical activity display a more favorable cardiometabolic biomarker profile than youths who engage in lesser amounts of moderate-vigorous physical activity. Intervention studies in obese youths show that favorable changes in biomarkers are produced by moderate-vigorous physical activity doses of 150-180 min/week. However, for nonobese youths, intervention studies suggest that such doses are not effective; higher moderate-vigorous physical activity doses of approximately 300 min/week seem necessary. Continuing a physically active lifestyle from childhood into the adult years will enable people to maintain less end-organ damage and lower rates of morbidity and mortality from cardiovascular disease and type 2 diabetes. PMID- 21633132 TI - Effects of strength training on motor performance skills in children and adolescents: a meta-analysis. AB - The recent literature delineates resistance training in children and adolescents to be effective and safe. However, only little is known about the transfer of achieved strength gains to athletic performance. The present meta-analysis revealed a combined mean effect size for motor skill types jumping, running, and throwing of 0.52 (95% CI: 0.33-0.71). Effect sizes for each of aforementioned skill types separately were 0.54 (95% CI: 0.34-0.74), 0.53 (95% CI: 0.23-0.83), and 0.99 (95% CI: 0.19-1.79) respectively. Furthermore, it could be shown that younger subjects and nonathletes showed higher gains in motor performance following resistance training than their counterparts and that specific resistance training regimes were not advantageous over traditional resistance training programs. Finally, a positive dose response relationship for "intensity" could be found in subgroups using traditional training regimens. These results emphasize that resistance training provides an effective way for enhancing motor performance in children and adolescents. PMID- 21633133 TI - Active commuting and physical activity in adolescents from Europe: results from the HELENA study. AB - We assessed commuting patterns in adolescents from 10 European cities and examined associations with physical activity (PA). A total of 3112 adolescents were included. PA was objectively measured with accelerometry. Commuting patterns and overall PA were self-reported using questions from the International Physical Activity Questionnaire modified for adolescents (IPAQ-A). Adolescents reported to spend 30 min (15,60) [expressed as median (25th, 75th percentiles)] walking. In boys, associations between active commuting (walking and biking) and PA levels were observed for moderate, moderate-to-vigorous and overall PA. In girls, these associations were observed for moderate and moderate-to-vigorous PA (walking). Similar results were found with the IPAQ-A. We observed positive associations between overall commuting and PA levels in European adolescents, yet due to the cross-sectional study design we cannot state the direction of these. Future studies should address the causation between active commuting and PA levels. PMID- 21633135 TI - Active travel to school and physical activity levels of Irish primary schoolchildren. AB - The purpose of this study was to (1) determine the physical activity levels of 9 11 year old children, and (2) compare the activity levels of children who commute to school by active and passive modes. 140 children aged 9-11 years (85 boys) were recruited from four urban Irish schools. Mode of commuting was assessed by questionnaire. Step counts were measured for 4 consecutive days. Mean daily step counts for the sample were 14386 +/- 5634. Boys were significantly more active than girls (15857 +/- 5482 vs. 12113 +/- 5127 steps). Eighty-seven children (62.1%) traveled by car, 51 children (36.4%) walked to school, one child traveled by bus and one child cycled. Children who walked or cycled to school had higher daily step counts than those who traveled by passive modes (16118 +/- 5757 vs. 13363 +/- 5332 steps). Active commuting to school may therefore represent a worthwhile strategy for improving children's physical activity levels. PMID- 21633134 TI - Association between light-intensity physical activity and adiposity in childhood. AB - The purpose of this study was to examine whether the association between daily light-intensity physical activity (LPA) and total body fat mass changes during childhood. The study sample was 577 children participating in the longitudinal Iowa Bone Development Study. Body fat mass and physical activity (PA) were measured using dual energy X-ray absorptiometry (DXA) and accelerometers, respectively, at approximately 5, 8, and 11 years of age. Age- and gender specific multivariable linear regression models were fit to predict fat mass by LPA, adjusted for actual age, birth weight, fat-free mass, height, moderate- to vigorous-intensity PA, and physical maturity (only for girls). Among boys, LPA was negatively associated with fat mass at age 11, but not age 5 or 8. Among girls, LPA was negatively associated with fat mass at ages 8 and 11, but not at age 5. LPA may have a beneficial effect against excess adiposity among older children. PMID- 21633136 TI - Longitudinal changes in physical self-perceptions and associations with physical activity during adolescence. AB - The purpose of this study was to examine adolescents' physical self-perceptions and their associations with physical activity using a longitudinal perspective. Utilizing data from the Physical Activity in Scottish Schoolchildren (PASS) study, changes in exercise self-efficacy, perceived competence, global self esteem and physical self-worth were assessed among a sample of 641 Scottish adolescents from age 11-15 years. Girls reported lower levels of perceived competence, self-esteem and physical self-worth than boys at each age. Furthermore, girls' physical self-perceptions decreased markedly over time. Among boys, only perceived competence decreased, while global self-esteem increased. Baseline physical activity was a significant predictor of later activity levels for both genders. Findings demonstrate the importance of physical self perceptions in relation to physical activity behavior among adolescents. Among older boys, high perceived competence increased the odds of being active by 3.8 times. Among older girls, high exercise self-efficacy increased the odds of being active by 5.2 times. There is a need for early interventions which promote increased physical literacy and confidence, particularly among girls. PMID- 21633137 TI - Reliability of fitness measures in 3- to 5-year-old children. AB - The purpose of this study was to determine the reliability of short-term power output, heart rate (HR) response during and after a treadmill test, and time to complete a 25-m dash in healthy preschool children. Thirty-two 3- to 5-year-old boys and girls completed two sessions approximately one week apart. Intraclass correlation coefficient (ICC) and coefficient of variation (CV) were calculated to evaluate reliability. Power output was found to be reliable with ICCs ranging from 0.83 to 0.93 and CVs from 8.1 to 9.7%. Time to complete the 25-m dash was highly reliable (ICC = 0.91, CV = 3.7%). Reliability for HR at submaximal exercise (ICC = 0.28, CV = 18.8%) and HR recovery (ICC = 0.42, CV = 14.0%) was not as strong. These findings should assist in determining appropriate fitness tests for preschoolers. PMID- 21633138 TI - Physical activity levels to estimate the energy requirement of adolescent athletes. AB - Adequate energy intake in adolescent athletes is considered important. Total energy expenditure (TEE) can be calculated from resting energy expenditure (REE) and physical activity level (PAL). However, validated PAL recommendations are available for adult athletes only. Purpose was to comprise physical activity data in adolescent athletes and to establish PAL recommendations for this population. In 64 competitive athletes (15.3 +/- 1.5 yr, 20.5 +/- 2.0 kg/m2) and 14 controls (15.1 +/- 1.1 yr, 21 +/- 2.1 kg/m2) TEE was calculated using 7-day activity protocols validated against doubly-labeled water. REE was estimated by Schofield HW equation, and PAL was calculated as TEE:REE. Observed PAL in adolescent athletes (1.90 +/- 0.35) did not differ compared with controls (1.84 +/- 0.32, p = .582) and was lower than recommended for adult athletes by the WHO. In conclusion, applicability of PAL values recommended for adult athletes to estimate energy requirements in adolescent athletes must be questioned. Instead, a PAL range of 1.75-2.05 is suggested. PMID- 21633139 TI - Clustering of metabolic syndrome risk factors associated with lifestyle factors and serum leptin in Korean children. AB - The present study investigated the relationships among metabolic risk factors, major lifestyle factors, and serum cytokines in a sample of Korean children. In a cross-sectional design, we studied a total of 275 children (130 boys and 145 girls) aged 12-13 years. Measured variables included anthropometrics, blood pressures (BP), VO2max, physical activity (PA), dietary intakes, lipids, glucose, and insulin. We explored the extent to which dietary intakes, VO2max, PA, and serum cytokines explained variance in a clustered risk score, which is a sum of Z scores for waist circumference, BP, TG, HDLC, and HOMA-IR, using a stepwise linear regression by blocks. VO2max, vigorous PA (VPA), and leptin were independent predictors for the clustered risk score while adjusting for age and Tanner stage. Our findings suggest that the clustered risk score is associated not only with low levels of VO2max and VPA, but also with elevated serum leptin in Korean children. PMID- 21633140 TI - Physical activity during soccer and its contribution to physical activity recommendations in normal weight and overweight children. AB - Amid the childhood obesity epidemic, understanding how organized sports participation contributes to meeting physical activity recommendations in children is important. Anthropometrics were measured in children (n = 111; 68% female, 9.1 +/- 0.8 yr) before one 50-min soccer match. Time spent at different physical activity intensity levels was examined using Actigraph accelerometers. 49% of the match time was spent in sedentary activity (25.4 +/- 5.7 min), while 33% of the match (16.9 +/- 4.7 min) was spent in moderate-to-vigorous activity (MVPA; p < .001). 22.5% of the children were overweight/obese and spent more time in sedentary activity (+3.2 +/- 1.2 min; p < .05) and less time in MVPA (-3.0 +/- 1.0 min; p < .01) compared with the normal weight children. These data demonstrate that playing an organized sport such as soccer only meets a portion (~25%) of the 60 min of MVPA recommended and even less of this recommendation is met by overweight/obese children. PMID- 21633141 TI - A critical review of "a randomized trial of nebulized 3% hypertonic saline with epinephrine in the treatment of acute bronchiolitis in the emergency department.". PMID- 21633143 TI - Rapid modulation of local neural activity by controlled drug release from polymer coated recording microelectrodes. AB - We demonstrate targeted perturbation of neuronal activity with controlled release of neurochemicals from conducting polymer-coated microelectrodes. Polymer coating and chemical incorporation are achieved through individually addressable electrodeposition, a process that does not compromise the recording capabilities of the electrodes. Release is realized by the application of brief voltage pulses that electrochemically reduce the polymer and dissociate incorporated neurochemicals; whereby they can diffuse away and achieve locally effective concentrations. Inhibition of evoked synaptic currents in neurons within 200 um of a 6-cyano-7-nitroquinoxaline-2,3-dione releasing electrode lasts for several seconds. Spiking activity of neurons in local circuits recorded extracellularly near the releasing electrode is silenced for a similar duration following release. This methodology is compatible with many neuromodulatory chemicals and various recording electrodes, including in vitro and implantable neural electrode arrays, thus providing an inexpensive and accessible technique capable of achieving sophisticated patterned chemical modulation of neuronal circuits. PMID- 21633146 TI - On the symmetry of wulfenite (Pb[MoO4]) from Meaeica (Slovenia). AB - Wulfenite [lead(II) molybdate(VI)] is known as a scheelite structure in the I4(1)/a space group. The structure of the unusual 'hemimorphic' wulfenite crystals from the Meaeica mine was refined in the noncentrosymmetric space group I 4 using a Pb/Mo exchange disorder model with the approximate composition Pb(0.94)Mo(0.06)[MoO(4)]. Pb atoms in the 2b positions are substituted by Mo at about 12%. The crystal is shown to be twinned by inversion. Hemimorphism may result from the short-range chemical ordering of the metal atoms at the 2b positions. PMID- 21633147 TI - Magnesium perchlorate anhydrate, Mg(ClO4)2, from laboratory X-ray powder data. AB - The previously unknown crystal structure of magnesium perchlorate anhydrate, determined and refined from laboratory X-ray powder diffraction data, represents a new structure type. The title compound was obtained by heating magnesium perchlorate hexahydrate at 523 K for 2 h under vacuum and it crystallizes in the monoclinic space group P2(1)/c. The asymmetric unit contains one Mg (site symmetry -1 on special position 2a), one Cl and four O sites (on general positions 4e). The structure consists of a three-dimensional network resulting from the corner-sharing of MgO(6) octahedra and ClO(4) tetrahedra. Each MgO(6) octahedron share corners with six ClO(4) tetrahedra. Each ClO(4) tetrahedron shares corners with three MgO(6) octahedra, with one O-atom corner dangling. The ClO(4) tetrahedra are oriented in such a way that one-dimensional channels parallel to [100] are formed between the dangling O atoms. PMID- 21633148 TI - Poly[(MU3-benzene-1,4-diacetato)[MU2-1,4-bis(1,2,4-triazol-1 yl)butane]cadmium(II)]: self assembly into a three-dimensional supramolecular framework based on [Cd(MU3-benzene-1,4-diacetate)] double chains. AB - The title compound, [Cd(C(10)H(8)O(4))(C(8)H(12)N(6))](n), crystallizes with an asymmetric unit comprising a divalent Cd(II) atom, a benzene-1,4-diacetate (PBEA(2-)) ligand and a complete 1,4-bis(1,2,4-triazol-1-yl)butane (BTB) ligand. [Cd(PBEA)](n) double chains, arranged parallel to the c axis, are formed through an exo-tridentate binding mode of the PBEA(2-) ligands. These [Cd(PBEA)](n) double chains are pillared by tethering BTB ligands, in which the BTB shows a trans-trans-trans conformation, to establish [Cd(PBEA)(BTB)](n) two-dimensional coordination polymer (4,4)-layer slab patterns. The three-dimensional supramolecular architecture is formed by C-H...O hydrogen bonds and C-H...pi interactions. PMID- 21633149 TI - Coordination-directed one-dimensional coordination polymers generated from a new oxadiazole bridging ligand and HgX2 (X=Cl, Br and I). AB - A new 1,3,4-oxadiazole bridging bent organic ligand, 2,5-bis{5-methyl-2-[(4 pyridyl)methoxy]phenyl}-1,3,4-oxadiazole, C(28)H(24)N(4)O(3), L, has been used to create three novel one-dimensional isomorphic coordination polymers, viz. catena poly[[[dichloridomercury(II)]-MU-2,5-bis{5-methyl-2-[(4-pyridyl)methoxy]phenyl} 1,3,4-oxadiazole] methanol monosolvate], {[HgCl(2)(C(28)H(24)N(4)O(3))].CH(3)OH}(n), catena-poly[[[dibromidomercury(II)] MU-2,5-bis{5-methyl-2-[(4-pyridyl)methoxy]phenyl}-1,3,4-oxadiazole] methanol monosolvate], {[HgBr(2)(C(28)H(24)N(4)O(3))].CH(3)OH}(n), and catena poly[[[diiodidomercury(II)]-MU-2,5-bis{5-methyl-2-[(4-pyridyl)methoxy]phenyl} 1,3,4-oxadiazole] methanol monosolvate], {[HgI(2)(C(28)H(24)N(4)O(3))].CH(3)OH}(n). The free L ligand itself adopts a cis conformation, with the two terminal pyridine rings and the central oxadiazole ring almost coplanar [dihedral angles = 5.994 (7) and 9.560 (6) degrees ]. In the Hg(II) complexes, however, one of the flexible pyridylmethyl arms of ligand L is markedly bent and helical chains are obtained. The Hg(II) atom lies in a distorted tetrahedral geometry defined by two pyridine N-atom donors from two L ligands and two halide ligands. The helical chains stack together via interchain pi-pi interactions that expand the dimensionality of the structure from one to two. The methanol solvent molecules link to the complex polymers through O-H...N and O-H...O hydrogen bonds. PMID- 21633151 TI - [CuCl3(H2O)](-) complexes aggregated to form hydrate columns in methyl substituted pyridinium or piperidinium salts. AB - 1,2,3-Trimethylpyridinium aquatrichloridocuprate(II), (C(8)H(12)N)[CuCl(3)(H(2)O)], (I), 3,4-dimethylpyridinium aquatrichloridocuprate(II), (C(7)H(10)N)[CuCl(3)(H(2)O)], (II), and 2,3 dimethylpyridinium aquatrichloridocuprate(II), (C(7)H(10)N)[CuCl(3)(H(2)O)], (III), exhibit the same fundamental structure, with (I) and (II) isomorphous and with the unit-cell constants of (III) similar to the reduced unit-cell constants of (I) and (II). The distorted square-planar [CuCl(3)(H(2)O)](-) complex [mirror symmetric in (I) and (II)] forms two semicoordinate Cu...Cl bonds to a neighboring complex to produce a dimer with 2/m symmetry [only inversion symmetry in (III)]. The semicoordinate Cu...Cl bond length of the dimer shows significant elongation at 295 K compared with that at 100 K, while the coordinate Cu-Cl bond lengths are slightly contracted at 295 K compared with those at 100 K. The inorganic dimers are linked by eight hydrogen bonds to four neighboring dimers to establish a checkerboard network layer in the ab plane, with voids between the dimers that accommodate, on both sides, inversion-related organic cation pairs. The organic cations are required by mirror-plane symmetry to be disordered in (I) and (II). The organic cations and [CuCl(3)(H(2)O)](-) complexes are nearly coplanar and tilted out of the layer plane to establish a hybrid organic inorganic layer structure parallel to (202) [(11-2) in (III)], with hydrate columns (defined by water molecules) and hydrophobic columns (defined by methyl groups) parallel to each other [and along the 2(1) axes in (I) and (II)]. In 1,1 dimethylpiperidinium aquatrichloridocuprate(II), (C(7)H(16)N)[CuCl(3)(H(2)O)], (IV), the bulkier organic cation prevents semicoordinate bonding between complexes, which are hydrogen bonded side-to-side in zigzag chains that place water molecules in columns along half of the 2(1) axes. PMID- 21633150 TI - MU-4,4'-Bipyridine-kappa2N:N'-bis[triaqua(4,4'-bipyridine-kappaN)cadmium(II)] bis(3-aminobenzoate) bis(perchlorate) dihydrate: a novel supramolecular system constructed by pi-pi and hydrogen-bonding interactions. AB - The title dicadmium compound, [Cd(2)(C(10)H(8)N(2))(5)(H(2)O)(6)](C(7)H(6)NO(2))(2)(ClO(4))(2).2H(2)O, is located around an inversion centre. Each Cd(II) centre is coordinated by three N atoms from three different 4,4'-bipyridine ligands and three O atoms from three coordinating water molecules in a distorted octahedral coordination environment. In the dicadmium cation unit, one 4,4'-bipyridine (4,4'-bipy) molecule acts as a bidentate bridging ligand between two Cd metal ions, while the other four 4,4' bipy molecules act only as monodentate terminal ligands, resulting in a rare 'H type' [Cd(2)(C(10)H(8)N(2))(5)(H(2)O)(6)] host unit. These host units are connected to each other via pi-pi stacking interactions, giving rise to a three dimensional supramolecular grid network with large cavities. The 3-aminobenzoate anions, perchlorate anions and water molecules are encapsulated in the cavities by numerous hydrogen-bonding interactions. To the best of our knowledge, this is the first example of a coordination compound based on both 4,4'-bipyridine ligands together with discrete 3-aminobenzoate anions. PMID- 21633152 TI - An X-ray powder diffraction study of cis-dichloridobis(2-methyl-2H-tetrazol-5 amine-kappaN(4))platinum(II), a tetrazole-containing analogue of cisplatin. AB - The structure of the title compound, cis-[PtCl(2)(C(2)H(5)N(5))(2)], was analysed using in-house X-ray powder diffraction data at room temperature. The structure was solved by direct methods and refined using Rietveld analysis. A slightly distorted square-planar coordination geometry is formed around the Pt atom by two Cl atoms and two ring N atoms of the 2-methyl-2H-tetrazol-5-amine ligands, which are in a cis configuration. The planes of the tetrazole rings are inclined at 79.7 (7) and 73.8 (6) degrees with respect to the coordination plane, with their substituents oriented in such a way that the complex as a whole has approximate C(2) symmetry. Intermolecular N-H...Cl hydrogen bonds mediate the formation of a three-dimensional supramolecular network. PMID- 21633153 TI - MU-2,2',6,6'-Tetramethyl-4,4'-bipyridine-kappa2N1:N1'-bis[(diethylenetriamine kappa3N,N',N'')palladium(II)] tetranitrate. AB - The title compound, [Pd(2)(C(4)H(13)N(3))(2)(C(14)H(16)N(2))](NO(3))(4), comprises discrete tetracationic dumbbell-type dinuclear complex molecules and noncoordinating nitrate anions. Two Pd(dien)(2+) moieties (dien is diethylenetriamine) are joined by the rigid linear exo-bidentate bridging 2,2',6,6'-tetramethyl-4,4'-bipyridine ligand to form the dinuclear complex, which lies across a centre of inversion in the space group P2(1)/n, so that the rings in the 2,2',6,6'-tetramethyl-4,4'-bipyridine bridging ligand are parallel. In the crystal, the primary and secondary amino groups of the dien ligand act as hydrogen-bond donors towards the nitrate anions to form a three-dimensional hydrogen-bond network. PMID- 21633154 TI - The pentacoordinated [Cr(CO)5](2-) dianion in [2,2,2-crypt-K]2[Cr(CO)5] ethylenediamine monosolvate. AB - Bis[(4,7,13,16,21,24-hexaoxa-1,10-diazabicyclo[8.8.8]hexacosane)potassium(+)] pentacarbonylchromate(2-) ethylenediamine monosolvate, [K(C(18)H(36)N(2)O(6))](2)[Cr(CO)(5)].C(2)H(8)N(2), was obtained from the reaction between K(3)Cd(2)Sb(2) and Cr(CO)(6) in ethylenediamine in the presence of the macrocyclic 2,2,2-crypt ligand. The structure provides the first crystallographic characterization of the pentacoordinated [Cr(CO)(5)](2-) dianion. The central Cr(III) atom is coordinated by five carbonyl ligands in a distorted trigonal-bipyramidal geometry. The distribution of the Cr-C bond lengths indicates a greater degree of back bonding from Cr(III) to the equatorial carbonyl ligands compared with the axial carbonyl ligands. PMID- 21633155 TI - Sodium tris(acetato-kappa2O,O')dioxidoamericate(VI) and guanidinium tris(cyclopropanecarboxylato-kappa2O,O')dioxidoamericate(VI). AB - The title compounds, Na[{AmO(2)}(C(2)H(3)O(2))(3)], (I), and (CH(6)N(3))[{AmO(2)}(C(4)H(5)O(2))(3)], (II), contain complex anions in which AmO(2)(2+) cations are surrounded by three bidentate-chelating carboxylate groups. The atoms of the AmO(2) group and the Na atoms in (I) are situated on threefold axes. All the atoms in (II) occupy general positions. Both compounds are isomorphous with earlier studied analogous compounds of previous members of the actinide (An) series. PMID- 21633156 TI - The diselanylbis(1,3-dimethyl-1H-imidazol-3-ium) dication stabilized by the polymeric catena-pentachloridotricuprate(I) anion. AB - In the title compound, catena-poly[diselanylbis(1,3-dimethyl-1H-imidazol-3-ium) [MU(3)-chlorido-tetra-MU(2)-chlorido-tricuprate(I)]], {(C(10)H(16)N(4)Se(2))[Cu(3)Cl(5)]}(n), the diselenide dication is stabilized by catena-[Cu(3)Cl(5)](2-) anions which associate through strong Cu-Cl bonds [average length = 2.3525 (13) A] to form polymeric chains. The polymeric [Cu(3)Cl(5)](2-) anion contains crystallographically imposed twofold rotation symmetry, with distorted trigonal-planar and tetrahedral geometries around the two symmetry-independent Cu atoms. Likewise, the Se-Se bond of the cation is centered on a twofold rotation axis. PMID- 21633157 TI - A novel three-dimensional heterometallic coordination polymer: poly[[diaquadi-MU3 chlorido-MU2-chlorido-tri-MU3-nicotinato-tricopper(I)gadolinium(III)] hemihydrate]. AB - In the title polymeric heterometallic compound, {[Cu(3)Gd(C(6)H(4)NO(2))(3)Cl(3)(H(2)O)(2)].0.5H(2)O}(n), comprising copper(I) and gadolinium(III) cations bridged by nicotinate (nic) ligands and chloride anions, the Gd(III) centers display a bicapped trigonal prismatic geometry, defined by six carboxylate O atoms and two water molecules. For copper(I), one Cu center is three-coordinated by three chloride ions and displays a trigonal-planar geometry; the other two Cu centers are four-coordinated and display a very distorted tetrahedral geometry. The chloride anions act in MU(2)- and MU(3) bridging modes, linking the Cu(I) ions into an infinite chain. The nic ligand exhibits a tridentate coordination mode, with the carboxylate O atoms linking to two Gd(III) ions and the N atom linking to one Cu(I) ion. Thus, a novel three dimensional heterometallic coordination polymer is constructed from Gd carboxylate subunits and Cu-Cl chains. In addition, intra- and intermolecular O H...O and O-H...Cl hydrogen bonds are also observed within the three-dimensional structure. Topologically, the framework represents an unusual 3,6-connected (4.8(2))(3)(4(10).6(5)) net. PMID- 21633158 TI - Two polymorphs of (2-carboxyethyl)(phenyl)phosphinic acid. AB - Two polymorphs of (2-carboxyethyl)(phenyl)phosphinic acid, C(9)H(11)O(4)P, crystallize in the chiral P2(1)2(1)2(1) space group with similar unit-cell parameters. They feature an essentially similar hydrogen-bonding motif but differ slightly in their detailed geometric parameters. For both polymorphs, the unequivocal location of the hydroxy H atoms together with the expected differences in the P-O bond lengths establish unequivocally that both forms contain the S isomer; the protonated phosphinic acid and carboxy O atoms serve as hydrogen-bond donors, while the second phosphinic acid O atom acts as a double hydrogen-bond acceptor and the remaining carboxy O atom is not involved in hydrogen bonding. Thus, an undulating two-dimensional supramolecular layer aggregate is formed based on an R(4)(3)(20) ring unit. Such polymorphism derives from the rotation of the C-C single bonds between the two hydrogen-bond-involved carboxy and phosphinic acid moieties. PMID- 21633159 TI - 1,3,5-Tris(bromomethyl)benzene. AB - The asymmetric unit of the title compound, C(9)H(9)Br(3), is composed of a single molecule. Two bromo substituents are located on one side of the plane of the aromatic ring and the third is on the opposite side, with the molecular unit exhibiting an approximate noncrystallographic C(s) point group. The crystal structure is rich in Br...Br, CH(2)...Br and CH...pi weak intermolecular contacts which mediate the crystal packing of individual molecules. These interactions promote a red-shift of a handful of vibrational modes (associated with the pendant -CH(2)Br groups) compared with values from theoretical density functional theory (DFT) calculations. PMID- 21633160 TI - Methyl 2-benzamido-4-(3,4-dimethoxyphenyl)-5-methylbenzoate and N-{5-benzoyl-2 [(Z)-2-methoxyethenyl]-4-methylphenyl}benzamide. AB - Methyl 2-benzamido-4-(3,4-dimethoxyphenyl)-5-methylbenzoate, C(24)H(23)NO(5), (Ia), and N-{5-benzoyl-2-[(Z)-2-methoxyethenyl]-4-methylphenyl}benzamide, C(24)H(21)NO(3), (IIa), were formed via a Diels-Alder reaction of appropriately substituted 2H-pyran-2-ones and methyl propiolate or (Z)-1-methoxybut-1-en-3-yne, respectively. Each of these cycloadditions might yield two different regioisomers, but just one was obtained in each case. In (Ia), an intramolecular N-H...O hydrogen bond closes a six-membered ring. A chain is formed due to aromatic pi-pi interactions, and a three-dimensional framework structure is formed by a combination of C-H...O and C-H...pi(arene) hydrogen bonds. Compound (IIa) was formed not only regioselectively but also chemoselectively, with just the triple bond reacting and the double bond remaining unchanged. Compound (IIa) crystallizes as N-H...O hydrogen-bonded dimers stabilized by aromatic pi-pi interactions. Dimers of (IIa) are connected into a chain by weak C-H...pi(arene) interactions. PMID- 21633161 TI - The molecular conformation of pentan-3-one studied in cholic acid pentan-3-one solvate. AB - The crystal structure of cholic acid-pentan-3-one (1/1), C(5)H(10)O.C(24)H(40)O(5), has been determined in order to deduce the molecular conformation of the small volatile ketone. Data were collected at 100 K to a resolution of (sin theta)/lambda = 0.91 A(-1). The structure contains a hydrogen bonded cholic acid host network, forming only van der Waals interactions with the guest pentan-3-one molecules. The ketone molecules are disordered on general positions, with two clearly identifiable conformations. The majority conformer exhibits approximate C(2) symmetry and is similar to that recently observed by microwave spectroscopy in the gas phase. PMID- 21633162 TI - 1-(Thiophen-3-yl)ethanone. AB - The structure of the title compound, C(6)H(6)OS, exhibits a flip-type disorder of the thiophene ring [occupancy ratio = 0.848 (3):0.152 (3)], which is typical for many thiophene derivatives. The puckered thiophene ring is essentially coplanar with the plane formed by the non-H atoms of the acetyl substituent, similar to its simple analogues, i.e. 3-acetyl-2-carboxythiophene, 4-acetyl-3 carboxythiophene and 3,5-diacetyl-2-ethylamino-4-methylthiophene. In the crystal structure, molecules are connected by C-H...pi hydrogen bonds, forming a sheet parallel to the (001) plane. Moreover, an inspection of the crystal lattice reveals that there are short S...O contacts connecting the molecules of adjacent sheets. Comparison of the title crystal structure with its simple 3 methoxythiophene analogue shows a close similarity in the herringbone arrangement of molecules and in the presence of C-H...pi interactions and S...O contacts. PMID- 21633163 TI - 1,5-Dianilinopentane-1,3,5-trione: a crystal structure containing two polymorphic domains. AB - Single crystals of the title compound, C(17)H(16)N(2)O(3), were obtained by gas diffusion. The observed diffraction pattern is compatible with a superposition of reflections from two monoclinic unit cells with the space group C2/c. The two cells share the a and b axes but not the c axis. Both structures contain layers parallel to (001), with molecules connected by intermolecular N-H...O=C hydrogen bonds. The bonding between adjacent layers is weak. Layer displacements result in a crystal structure containing two closely related polymorphic domains. The structure of one polymorph can be derived from the structure of the other if subsequent layers are displaced by (a/4, b/4, 0) for odd-numbered layers and by (a/4, -b/4, 0) for even-numbered layers. Three different crystals were analysed and their observed diffraction patterns were similar, showing all three crystals to contain the polymorphic domain structure. PMID- 21633164 TI - Mutations in KIF7 link Joubert syndrome with Sonic Hedgehog signaling and microtubule dynamics. AB - Joubert syndrome (JBTS) is characterized by a specific brain malformation with various additional pathologies. It results from mutations in any one of at least 10 different genes, including NPHP1, which encodes nephrocystin-1. JBTS has been linked to dysfunction of primary cilia, since the gene products known to be associated with the disorder localize to this evolutionarily ancient organelle. Here we report the identification of a disease locus, JBTS12, with mutations in the KIF7 gene, an ortholog of the Drosophila kinesin Costal2, in a consanguineous JBTS family and subsequently in other JBTS patients. Interestingly, KIF7 is a known regulator of Hedgehog signaling and a putative ciliary motor protein. We found that KIF7 co-precipitated with nephrocystin-1. Further, knockdown of KIF7 expression in cell lines caused defects in cilia formation and induced abnormal centrosomal duplication and fragmentation of the Golgi network. These cellular phenotypes likely resulted from abnormal tubulin acetylation and microtubular dynamics. Thus, we suggest that modified microtubule stability and growth direction caused by loss of KIF7 function may be an underlying disease mechanism contributing to JBTS. PMID- 21633165 TI - The JAK2/STAT3 signaling pathway is required for growth of CD44+CD24- stem cell like breast cancer cells in human tumors. AB - Intratumor heterogeneity is a major clinical problem because tumor cell subtypes display variable sensitivity to therapeutics and may play different roles in progression. We previously characterized 2 cell populations in human breast tumors with distinct properties: CD44+CD24- cells that have stem cell-like characteristics, and CD44-CD24+ cells that resemble more differentiated breast cancer cells. Here we identified 15 genes required for cell growth or proliferation in CD44+CD24- human breast cancer cells in a large-scale loss-of function screen and found that inhibition of several of these (IL6, PTGIS, HAS1, CXCL3, and PFKFB3) reduced Stat3 activation. We found that the IL-6/JAK2/Stat3 pathway was preferentially active in CD44+CD24- breast cancer cells compared with other tumor cell types, and inhibition of JAK2 decreased their number and blocked growth of xenografts. Our results highlight the differences between distinct breast cancer cell types and identify targets such as JAK2 and Stat3 that may lead to more specific and effective breast cancer therapies. PMID- 21633166 TI - Identification of human triple-negative breast cancer subtypes and preclinical models for selection of targeted therapies. AB - Triple-negative breast cancer (TNBC) is a highly diverse group of cancers, and subtyping is necessary to better identify molecular-based therapies. In this study, we analyzed gene expression (GE) profiles from 21 breast cancer data sets and identified 587 TNBC cases. Cluster analysis identified 6 TNBC subtypes displaying unique GE and ontologies, including 2 basal-like (BL1 and BL2), an immunomodulatory (IM), a mesenchymal (M), a mesenchymal stem-like (MSL), and a luminal androgen receptor (LAR) subtype. Further, GE analysis allowed us to identify TNBC cell line models representative of these subtypes. Predicted "driver" signaling pathways were pharmacologically targeted in these cell line models as proof of concept that analysis of distinct GE signatures can inform therapy selection. BL1 and BL2 subtypes had higher expression of cell cycle and DNA damage response genes, and representative cell lines preferentially responded to cisplatin. M and MSL subtypes were enriched in GE for epithelial-mesenchymal transition, and growth factor pathways and cell models responded to NVP-BEZ235 (a PI3K/mTOR inhibitor) and dasatinib (an abl/src inhibitor). The LAR subtype includes patients with decreased relapse-free survival and was characterized by androgen receptor (AR) signaling. LAR cell lines were uniquely sensitive to bicalutamide (an AR antagonist). These data may be useful in biomarker selection, drug discovery, and clinical trial design that will enable alignment of TNBC patients to appropriate targeted therapies. PMID- 21633167 TI - CCL17-expressing dendritic cells drive atherosclerosis by restraining regulatory T cell homeostasis in mice. AB - Immune mechanisms are known to control the pathogenesis of atherosclerosis. However, the exact role of DCs, which are essential for priming of immune responses, remains elusive. We have shown here that the DC-derived chemokine CCL17 is present in advanced human and mouse atherosclerosis and that CCL17+ DCs accumulate in atherosclerotic lesions. In atherosclerosis-prone mice, Ccl17 deficiency entailed a reduction of atherosclerosis, which was dependent on Tregs. Expression of CCL17 by DCs limited the expansion of Tregs by restricting their maintenance and precipitated atherosclerosis in a mechanism conferred by T cells. Conversely, a blocking antibody specific for CCL17 expanded Tregs and reduced atheroprogression. Our data identify DC-derived CCL17 as a central regulator of Treg homeostasis, implicate DCs and their effector functions in atherogenesis, and suggest that CCL17 might be a target for vascular therapy. PMID- 21633168 TI - FGF-dependent regulation of VEGF receptor 2 expression in mice. AB - Numerous studies have suggested a link between the angiogenic FGF and VEGF signaling pathways; however, the nature of this link has not been established. To evaluate this relationship, we investigated VEGF signaling in ECs with disrupted FGF signaling in vitro and in vivo. ECs lacking FGF signaling became unresponsive to VEGF, caused by downregulation of VEGF receptor 2 (VEGFR2) expression after reduced Vegfr2 enhancer activation. FGF mediated VEGFR2 expression via activation of Erk1/2. Transcriptional analysis revealed that Ets transcription factors controlled VEGFR2 expression in an FGF- and Erk1/2-dependent manner. Mice with defective FGF signaling exhibited loss of vascular integrity and reduced vascular morphogenesis. Thus, basal FGF stimulation of the endothelium is required for maintenance of VEGFR2 expression and the ability to respond to VEGF stimulation and accounts for the hierarchic control of vascular formation by FGFs and VEGF. PMID- 21633169 TI - Loss of Gata5 in mice leads to bicuspid aortic valve. AB - Bicuspid aortic valve (BAV), the leading congenital heart disease, occurs in 1% 2% of the population. Genetic studies suggest that BAV is an autosomal-dominant disease with reduced penetrance. However, only 1 gene, NOTCH1, has been linked to cases of BAV. Here, we show that targeted deletion of Gata5 in mice leads to hypoplastic hearts and partially penetrant BAV formation. Endocardial cell specific inactivation of Gata5 led to BAV, similar to that observed in Gata5-/- mice. In all cases, the observed BAVs resulted from fusion of the right-coronary and noncoronary leaflets, the subtype associated with the more severe valve dysfunction in humans. Neither endocardial cell proliferation nor cushion formation was altered in the absence of Gata5. Rather, defective endocardial cell differentiation, resulting from the deregulation of several components of the Notch pathway and other important endocardial cell regulators, may be the underlying mechanism of disease. The results unravel a critical cell-autonomous role for endocardial Gata5 in aortic valve formation and identify GATA5 as a potential gene responsible for congenital heart disease in humans. Mice with mutated Gata5 alleles represent unique models to dissect the mechanisms underlying degenerative aortic valve disease and to develop much-needed preventive and therapeutic interventions. PMID- 21633170 TI - Inhibition of PKCdelta reduces cisplatin-induced nephrotoxicity without blocking chemotherapeutic efficacy in mouse models of cancer. AB - Cisplatin is a widely used cancer therapy drug that unfortunately has major side effects in normal tissues, notably nephrotoxicity in kidneys. Despite intensive research, the mechanism of cisplatin-induced nephrotoxicity remains unclear, and renoprotective approaches during cisplatin-based chemotherapy are lacking. Here we have identified PKCdelta as a critical regulator of cisplatin nephrotoxicity, which can be effectively targeted for renoprotection during chemotherapy. We showed that early during cisplatin nephrotoxicity, Src interacted with, phosphorylated, and activated PKCdelta in mouse kidney lysates. After activation, PKCdelta regulated MAPKs, but not p53, to induce renal cell apoptosis. Thus, inhibition of PKCdelta pharmacologically or genetically attenuated kidney cell apoptosis and tissue damage, preserving renal function during cisplatin treatment. Conversely, inhibition of PKCdelta enhanced cisplatin-induced cell death in multiple cancer cell lines and, remarkably, enhanced the chemotherapeutic effects of cisplatin in several xenograft and syngeneic mouse tumor models while protecting kidneys from nephrotoxicity. Together these results demonstrate a role of PKCdelta in cisplatin nephrotoxicity and support targeting PKCdelta as an effective strategy for renoprotection during cisplatin-based cancer therapy. PMID- 21633171 TI - S6 kinase 1 is required for rapamycin-sensitive liver proliferation after mouse hepatectomy. AB - Rapamycin is an antibiotic inhibiting eukaryotic cell growth and proliferation by acting on target of rapamycin (TOR) kinase. Mammalian TOR (mTOR) is thought to work through 2 independent complexes to regulate cell size and cell replication, and these 2 complexes show differential sensitivity to rapamycin. Here we combine functional genetics and pharmacological treatments to analyze rapamycin-sensitive mTOR substrates that are involved in cell proliferation and tissue regeneration after partial hepatectomy in mice. After hepatectomy, hepatocytes proliferated rapidly, correlating with increased S6 kinase phosphorylation, while treatment with rapamycin derivatives impaired regeneration and blocked S6 kinase activation. In addition, genetic deletion of S6 kinase 1 (S6K1) caused a delay in S phase entry in hepatocytes after hepatectomy. The proliferative defect of S6K1 deficient hepatocytes was cell autonomous, as it was also observed in primary cultures and hepatic overexpression of S6K1-rescued proliferation. We found that S6K1 controlled steady-state levels of cyclin D1 (Ccnd1) mRNA in liver, and cyclin D1 expression was required to promote hepatocyte cell cycle. Notably, in vivo overexpression of cyclin D1 was sufficient to restore the proliferative capacity of S6K-null livers. The identification of an S6K1-dependent mechanism participating in cell proliferation in vivo may be relevant for cancer cells displaying high mTOR complex 1 activity and cyclin D1 accumulation. PMID- 21633172 TI - Attenuation of HIV-associated human B cell exhaustion by siRNA downregulation of inhibitory receptors. AB - Chronic immune activation in HIV-infected individuals leads to accumulation of exhausted tissue-like memory B cells. Exhausted lymphocytes display increased expression of multiple inhibitory receptors, which may contribute to the inefficiency of HIV-specific antibody responses. Here, we show that downregulation of B cell inhibitory receptors in primary human B cells led to increased tissue-like memory B cell proliferation and responsiveness against HIV. In human B cells, siRNA knockdown of 9 known and putative B cell inhibitory receptors led to enhanced B cell receptor-mediated (BCR-mediated) proliferation of tissue-like memory but not other B cell subpopulations. The strongest effects were observed with the putative inhibitory receptors Fc receptor-like-4 (FCRL4) and sialic acid-binding Ig-like lectin 6 (Siglec-6). Inhibitory receptor downregulation also led to increased levels of HIV-specific antibody-secreting cells and B cell-associated chemokines and cytokines. The absence of known ligands for FCRL4 and Siglec-6 suggests these receptors may regulate BCR signaling through their own constitutive or tonic signaling. Furthermore, the extent of FCLR4 knockdown effects on BCR-mediated proliferation varied depending on the costimulatory ligand, suggesting that inhibitory receptors may engage specific pathways in inhibiting B cell proliferation. These findings on HIV associated B cell exhaustion define potential targets for reversing the deleterious effect of inhibitory receptors on immune responses against persistent viral infections. PMID- 21633173 TI - The three R's of lung health and disease: repair, remodeling, and regeneration. AB - All tissues and organs can be classified according to their ability to repair and regenerate during adult homeostasis and after injury. Some exhibit a high rate of constant cell turnover, while others, such as the lung, exhibit only low-level cell regeneration during normal adult homeostasis but have the ability to rapidly regenerate new cells after injury. Lung regeneration likely involves both activation of progenitor cells as well as cell replacement through proliferation of remaining undamaged cells. The pathways and factors that control this process and its role in disease are only now being explored. In this Review, we will discuss the connection between pathways required for lung development and how the lung responds to injury and disease, with a particular emphasis on recent studies describing the role for the epithelium in repair and regeneration. PMID- 21633174 TI - Digging deeper into obesity. AB - The growing problem of obesity is associated with multiple morbidities, including increased risk of diabetes, hypertension, heart disease, sleep apnea, and cancer. Obesity promotes disability, decreases productivity, and shortens life span. Although much attention has been focused on diet and exercise, these strategies alone are not effective in preventing obesity and maintaining weight loss. Moreover, the development of pharmacological approaches for obesity treatment has been dogged by poor efficacy and serious side effects. The biology of obesity is very complex, and mechanisms linking obesity to various diseases are poorly understood. This issue of the JCI highlights important concepts in our understanding of the pathogenesis of obesity and its complications. PMID- 21633175 TI - Genetic approaches to understanding human obesity. AB - Obesity and its associated comorbidities represent one of the biggest public health challenges facing the world today. The heritability of body weight is high, and genetic variation plays a major role in determining the interindividual differences in susceptibility or resistance to the obesogenic environment. Here we discuss how genetic studies in humans have contributed to our understanding of the central pathways that govern energy homeostasis. We discuss how the arrival of technological advances such as next-generation sequencing will result in a major acceleration in the pace of gene discovery. The study of patients harboring these genetic variants has informed our understanding of the molecular and physiological pathways involved in energy homeostasis. We anticipate that future studies will provide the framework for the development of a more rational targeted approach to the prevention and treatment of genetically susceptible individuals. PMID- 21633176 TI - Sixteen years and counting: an update on leptin in energy balance. AB - Cloned in 1994, the ob gene encodes the protein hormone leptin, which is produced and secreted by white adipose tissue. Since its discovery, leptin has been found to have profound effects on behavior, metabolic rate, endocrine axes, and glucose fluxes. Leptin deficiency in mice and humans causes morbid obesity, diabetes, and various neuroendocrine anomalies, and replacement leads to decreased food intake, normalized glucose homeostasis, and increased energy expenditure. Here, we provide an update on the most current understanding of leptin-sensitive neural pathways in terms of both anatomical organization and physiological roles. PMID- 21633177 TI - Adipose tissue remodeling and obesity. AB - To fulfill its role as the major energy-storing tissue, adipose has several unique properties that cannot be seen in any other organ, including an almost unlimited capacity to expand in a non-transformed state. As such, the tissue requires potent mechanisms to remodel, acutely and chronically. Adipocytes can rapidly reach the diffusional limit of oxygen during growth; hypoxia is therefore an early determinant that limits healthy expansion. Proper expansion requires a highly coordinated response among many different cell types, including endothelial precursor cells, immune cells, and preadipocytes. There are therefore remarkable similarities between adipose expansion and growth of solid tumors, a phenomenon that presents both an opportunity and a challenge, since pharmacological interventions supporting healthy adipose tissue adaptation can also facilitate tumor growth. PMID- 21633178 TI - The role of lipid droplets in metabolic disease in rodents and humans. AB - Lipid droplets (LDs) are intracellular organelles that store neutral lipids within cells. Over the last two decades there has been a dramatic growth in our understanding of LD biology and, in parallel, our understanding of the role of LDs in health and disease. In its simplest form, the LD regulates the storage and hydrolysis of neutral lipids, including triacylglycerol and/or cholesterol esters. It is becoming increasingly evident that alterations in the regulation of LD physiology and metabolism influence the risk of developing metabolic diseases such as diabetes. In this review we provide an update on the role of LD associated proteins and LDs in metabolic disease. PMID- 21633179 TI - Inflammatory links between obesity and metabolic disease. AB - The obesity epidemic has forced us to evaluate the role of inflammation in the health complications of obesity. This has led to a convergence of the fields of immunology and nutrient physiology and the understanding that they are inextricably linked. The reframing of obesity as an inflammatory condition has had a wide impact on our conceptualization of obesity-associated diseases. In this Review, we highlight the cellular and molecular mechanisms at play in the generation of obesity-induced inflammation. We also emphasize how defining the immune regulation in metabolic tissues has broadened the understanding of the diversity of inflammatory responses. PMID- 21633180 TI - Dynamics of insulin secretion and the clinical implications for obesity and diabetes. AB - Insulin secretion is a highly dynamic process regulated by various factors including nutrients, hormones, and neuronal inputs. The dynamics of insulin secretion can be studied at different levels: the single beta cell, pancreatic islet, whole pancreas, and the intact organism. Studies have begun to analyze cellular and molecular mechanisms underlying dynamics of insulin secretion. This review focuses on our current understanding of the dynamics of insulin secretion in vitro and in vivo and discusses their clinical relevance. PMID- 21633181 TI - Gut microbiome, obesity, and metabolic dysfunction. AB - The prevalence of obesity and related disorders such as metabolic syndrome has vastly increased throughout the world. Recent insights have generated an entirely new perspective suggesting that our microbiota might be involved in the development of these disorders. Studies have demonstrated that obesity and metabolic syndrome may be associated with profound microbiotal changes, and the induction of a metabolic syndrome phenotype through fecal transplants corroborates the important role of the microbiota in this disease. Dietary composition and caloric intake appear to swiftly regulate intestinal microbial composition and function. As most findings in this field of research are based on mouse studies, the relevance to human biology requires further investigation. PMID- 21633183 TI - Time-course analysis of DNA damage response-related genes after in vitro radiation in H460 and H1229 lung cancer cell lines. AB - Radiation is the most useful treatment modality for cancer patients. It initiates a series of signal cascades such as DNA damage response (DDR) signaling for repairing damaged DNA, arresting the cell cycle, and inducing cell death. Until now, few genes have been found to be regulated by radiation, which explains the molecular mechanisms of cellular responses to radiation. Although the transcriptional changes caused by radiation have been widely investigated, little is known about the direct evidence for the transcriptional control of DDR-related genes. Here, we examined the radiosensitivity of two non-small cell lung cancer cell lines (H460 and H1299), which have different p53 status. We monitored the time-dependent changes of 24 DDR-related gene expressions via microarray analysis. Based on the basal expression levels and temporal patterns, we further classified 24 DDR-related genes into four subgroups. Then, we also addressed the protein levels of several DDR-related genes such as TopBP1, Chk1 and Chk2, confirming the results of microarray analysis. Together, these results indicate that the expression patterns of DDR-related genes are associated with radiosensitivity and with the p53 statuses of H460 and H1299, which adds to the understanding of the complex biological responses to radiation. PMID- 21633184 TI - Inhibitory effect of (-)-epigallocatechin gallate on titanium particle-induced TNF-alpha release and in vivo osteolysis. AB - Tumor necrosis factor-alpha (TNF-alpha) and inflammatory cytokines released from activated macrophages in response to particulate debris greatly impact periprosthetic bone loss and consequent implant failure. In the present study, we found that a major polyphenolic component of green tea, (-)-epigallocatechin gallate (EGCG), inhibited Ti particle-induced TNF-alpha release in macrophages in vitro and calvarial osteolysis in vivo. The Ti stimulation of macrophages released TNF-alpha in a dose- and time-dependent manner, and EGCG substantially suppressed Ti particle-induced TNF-alpha release. Analysis of signaling pathway showed that EGCG inhibited the Ti-induced c-Jun N-terminus kinase (JNK) activation and inhibitory kappaB (IkappaB) degradation, and consequently the Ti induced transcriptional activation of AP-1 and NF-kappaB. In a mouse calvarial osteolysis model, EGCG inhibited Ti particle-induced osteolysis in vivo by suppressing TNF-a expression and osteoclast formation. Therefore, EGCG may be a potential candidate compound for osteolysis prevention and treatment as well as aseptic loosening after total replacement arthroplasty. PMID- 21633182 TI - Circadian rhythms, sleep, and metabolism. AB - The discovery of the genetic basis for circadian rhythms has expanded our knowledge of the temporal organization of behavior and physiology. The observations that the circadian gene network is present in most living organisms from eubacteria to humans, that most cells and tissues express autonomous clocks, and that disruption of clock genes results in metabolic dysregulation have revealed interactions between metabolism and circadian rhythms at neural, molecular, and cellular levels. A major challenge remains in understanding the interplay between brain and peripheral clocks and in determining how these interactions promote energy homeostasis across the sleep-wake cycle. In this Review, we evaluate how investigation of molecular timing may create new opportunities to understand and develop therapies for obesity and diabetes. PMID- 21633185 TI - Islet amyloid polypeptide in pancreatic islets from type 1 diabetic subjects. AB - AIMS/HYPOTHESIS: Islet amyloid polypeptide is originally isolated as the chief constituent of amyloid deposits in type 2 diabetic islets. Islet amyloid polypeptide hyposecretion was known in type 1 diabetics and this study aimed to detect possibly reduced islet amyloid polypeptide-positive cells in type 1 diabetic islets. RESULTS: Non-diabetic control islets showed about 60% of islet cells were insulin cells, and 60% of insulin cells were positive for IAPP. In type 1 diabetic islets, islets were generally smaller than control islets, consisting of weaker positive cells for insulin and islet amyloid polypeptide. Medium-sized islets still retained some insulin positive cells, whereas islet amyloid polypeptide positive cells were much less or even absent, but some insulin-negative cells were weakly islet amyloid polypeptide positive. An occasional extra-large islet, representing regenerating islets, consisting of more than 100 islet cells revealed less than 35% insulin and 20% islet amyloid polypeptide positive cells with relatively increased glucagon and somatostatin cells. Both normal and type 1 diabetic islets revealed scattered, densely insulin and islet amyloid polypeptide positive sickle-shaped cytoplasm without granular appearance, consistent with degenerating insulin cells. METHODS: Using commercially available rabbit anti-islet amyloid polypeptide antibody, immunostaning was performed on ten cases of type 1 diabetic pancreata and eight non-diabetic controls. Both control and type 1 diabetic pancreata were systematically immunostained for insulin, glucagon, somatostatin and islet amyloid polypeptide. CONCLUSION/INTERPRETATION: Control islets consisted of about 60% insulin cells, and about 34% of islet cells were amyloid polypeptide positive with scattered and densely positive for insulin and islet amyloid polypeptide without granular appearance, consistent with degenerating beta cells. All islets, including occasional extra-large islets from type 1 diabetics, showed less insulin cells and less islet amyloid polypeptide positive cells with twice increased glucagon and somatostatin cells of the control islets, but some insulin negative cells were positive for islet amyloid polypeptide, suggesting the presence of islet amyloid polypeptide in degenerating and extra-large regenerating islets. Thus, this immunocytochemical staining revealed generally less islet amyloid positive cells in type 1 diabetic islets, corresponding to severe hyposecretion of islet amyloid polypeptide in type 1 diabetics. PMID- 21633186 TI - Site-specific PEGylation endows a mammalian ribonuclease with antitumor activity. AB - Mammalian ribonucleases are emerging as cancer chemotherapeutic agents. Their cationicity engenders cell permeability, and their enzymatic activity destroys the biochemical information encoded by RNA. The pharmacologic potential of ribonucleases is, however, obviated by their high sensitivity to a cytosolic inhibitor protein (RI) and their small size, which limits their residence in serum. We reasoned that site specific conjugation of a poly(ethylene glycol) (PEG) chain could both reduce sensitivity to RI and increase serum half-life. We found that appending a PEG moiety can enable bovine pancreatic ribonuclease (RNase A) to evade RI, depending on the site of conjugation and the length and branching of the chain. Although a pendant PEG moiety decreases antiproliferative activity in vitro, PEGylation discourages renal clearance in vivo and leads to nearly complete tumor growth inhibition in a mouse xenograft model. These data demonstrate that a pendant PEG moiety can be beneficial to the action of proteins that act within the cytosol, and that strategic site-specific PEGylation can endow a mammalian ribonuclease with potent antitumor activity. PMID- 21633187 TI - Kinetics and genomic profiling of adult human and mouse beta-cell maturation. AB - Diabetes is a multifactorial metabolic disorder defined by the loss of functional pancreatic insulin-producing beta-cells. The functional maturation and dedifferentiation of adult beta-cells is central to diabetes pathogenesis and to beta-cell replacement therapy for the treatment of diabetes. Despite its importance, the dynamics and mechanisms of adult beta-cell maturation remain poorly understood. Using a novel Pdx1/Ins1 dual fluorescent reporter lentiviral vector, we previously found that individual adult human and mouse beta-cells exist in at least two differentiation states distinguishable by the activation of the rat Ins1 promoter and performed the first real-time imaging of the maturation of individual cultured beta-cells. Our previous study focused on transformed (MIN6) beta-cells as a model to investigatethe kinetics of beta-cell maturation. In the present study, we investigated the kinetics of the maturation process in primary human and mouse beta-cells and performed gene expression profiling. Gene expression profiling of FACS purified immature Pdx1 (+) /Ins1 (low) cells and mature Pdx1 (high) /Ins1 (high ) cells from cultures of human islets, mouse islets and MIN6 cells revealed that Pdx1 (+) /Ins1 (low) cells are enriched for multiple genes associated with beta-cell development/progenitor cells, proliferation, apoptosis, as well as genes coding for other islet cell hormones such as glucagon. We also demonstrated that the heterogeneity in beta-cell maturation states previously observed in vitro, can also be found in vivo. Collectively, these experiments contribute to the understanding of maturation, dedifferentiation and plasticity of adult pancreatic beta-cells. The results have significant implications for islet regeneration and for in vitro generation of functional beta-cells to treat diabetes. PMID- 21633188 TI - Dual roles for FY in the regulation of FLC. AB - In Arabidopsis, the flowering decision is determined by multiple pathways that integrate information from both endogenous signals and environmental cues. The genes of the autonomous pathway promote flowering by suppressing the expression of the floral repressor FLOWERING LOCUS C (FLC). Thus, autonomous-pathway mutants have elevated levels of FLC and are late flowering. Previous work has shown that two autonomous pathway proteins, FCA and FY, physically interact and this interaction is important in the repression of FLC. Recent work from our laboratory has shown that a hypomorphic allele of FY (fy-5) can cause earlier or later flowering, depending on the genetic background. These results suggest that FY has the potential to act as both an activator and a repressor of FLC. The FLC activating activity of FY appears to be FCA-independent, as fy-5 causes earlier flowering in an fca-null background. Here we present a speculative model that reconciles these opposing phenotypes by proposing a dual role for FY in the regulation of flowering time. PMID- 21633189 TI - N-acylethanolamine (NAE) inhibits growth in Arabidopsis thaliana seedlings via ABI3-dependent and -independent pathways. AB - N-acylethanolamines (NAEs) are lipid metabolites derived from the hydrolysis of the membrane phospholipid N-acylphosphatidylethanolamine (NAPE). Recent work in Arabidopsis thaliana seedlings showed that combined treatments of NAE 12:0 and ABA inhibited seedling growth synergistically, suggesting low levels of NAE could potentiate the action of ABA. Here we examined the interplay between compound concentrations, growth inhibition and mutant genotypes with impaired sensitivities to these regulators. NAE 12:0 and ABA both induced dose-dependent increases in transcript levels of ABI3, and two ABI3 responsive genes, AtHVA22B and RD29B. Interestingly, even in the absence of growth inhibition, RD29B transcripts were elevated by ABA but not NAE treatment outside the sensitive window for ABA/NAE treatment, indicating some differences in the regulation of growth and the modulation of gene expression by these two compounds. Also noteworthy, the growth of ABA insensitive mutant (abi 3-1) seedlings was inhibited at higher concentrations of NAE 12:0 but not ABA, suggesting that NAE may act to inhibit early seedling establishment by both ABI3-dependent and ABI3 independent pathways. Collectively our results reinforce the concept that NAE12:0 interacts with ABA signaling in seedling establishment, but also points to a complexity in this interaction that modulates the sensitivity of young seedlings to phytohormone-mediated growth arrest. PMID- 21633190 TI - Circadian rhythm leaf movement of Phaseolus vulgaris and the role of calcium ions. AB - Legume plants, due to their distinctive botanical characteristics, such as leaf movements, physiological characteristics, such as nitrogen fixation, and their abilities to endure environmental stresses, have important roles in sustainable pastures development. Leaf movement of legume plants is turgor regulated and osmotically active fluxes of ions between extensor and flexor of pulvinus cause this movement. To determine the role of calcium ions in circadian leaf movements of Phaseolus vulgaris L., a radiotracer technique experiment using 45Ca ions were employed. Measurements were taken during circadian leaf movements, and samples were taken from different parts of the leaflet. The 45Ca beta-particle activity reduced from leaflet base pulvinus to leaf tip. The pulvinus had the highest activity, while the leaf tip had the lowest. By increase of the ratio of 45Ca beta-particle activity within flexor to extensor (Fl/Ex) the midrib-petiole angle, as an indicator of leaf movement, increased linearly during circadian leaf movement (r = 0.86). The 45Ca beta-particle activity of Flex/Ext ratio reduced linearly (r = -0.88) toward midnight. In conclusion, it was found that calcium ions accumulation is opposite to the fluxes of osmatically active ions and water movement. Calcium ions accumulate at less negative water potential side of the pulivnus. PMID- 21633191 TI - Air pollution impedes plant-to-plant communication, but what is the signal? AB - Since the first reports that undamaged plants gain defensive benefits following exposure to damaged neighbors, the idea that plants may signal to each other has attracted much interest. There has also been substantial debate concerning the ecological significance of the process and the evolutionary drivers. Part of this debate has centered on the distance over which signaling between plants occurs in nature. In a recent study we showed that an ozone concentration of 80 ppb, commonly encountered in nature, significantly reduces the distance over which plant-plant signaling occurs in lima bean. We went on to show that degradation of herbivore-induced plant volatiles by ozone is the likely mechanism for this. The key question remaining from our work was that if ozone is degrading the signal in transit between plants, which chemicals are responsible for transmitting the signal in purer air? Here we present the results of a small scale experiment testing the role of the two most significant herbivore-induced terpenes and discuss our results in terms of other reported functions for these chemicals in plant-plant signaling. PMID- 21633192 TI - The mechanism of substrate recognition of Ca2+-dependent protein kinases. AB - Ca2+-dependent protein kinases (CDPKs) are encoded by a multigene family and are thought to play central roles in Ca2+ signaling in plants. Although the primary structures of CDPK isoforms are highly conserved, several studies suggested a distinct physiological function for each CDPK isoform in plants. Hence, there should be mechanisms by which individual CDPK specifically recognizes its substrate. Recently, the variable N-terminal domain of NtCDPK1 was shown to play an essential role in the specific recognition of the substrate. Because the variable N-terminal domain of other CDPKs may also be involved in the substrate recognition, the search for interacting proteins of the variable N-terminal domain would provide important clues to identify the physiological substrates of each CDPK. Additionally, manipulation of the variable N-terminal domain may enable us to engineer the substrate specificity of CDPK, leading a rational rewiring of cellular signaling pathways. PMID- 21633193 TI - The Tic20 gene family: phylogenetic analysis and evolutionary considerations. AB - Tic20 is a polytopic protein of the inner envelope membrane of chloroplasts, and it is proposed to act as a translocation channel during chloroplast protein import. By analysing 29 sequences from diverse organisms, it was evident that Tic20-related proteins form two distinct clades, termed Group 1 and Group 2. The former group includes canonical Tic20 proteins that are essential for chloroplast development, while members of the latter are of unknown function. An increased evolutionary rate, in connection with adaptation to terrestrial life, was detected in Group 1. Interestingly, the sub-cellular (genomic) localization of genes coding for Group 1 proteins differs between evolutionary lineages. PMID- 21633194 TI - Streptozotocin is responsible for the induction and progression of renal tumorigenesis in diabetic Wistar-Furth rats treated with insulin or transplanted with agarose encapsulated porcine islets. AB - Streptozotocin (STZ), a nitrosourea with DNA alkylating properties, has been widely used to induce hyperglycemia by specifically destroying the insulin producing beta-cells of the islets of Langerhans in experimental models of Type I diabetes. STZ's known carcinogenic properties, however, raise concerns about its suitability for long-term studies. We conducted a formal study of STZ's carcinogenic effects in long-term surviving diabetic Wistar-Furth rats. To determine if insulin therapy or islet transplantation exacerbated tumorigenesis, rats were randomly assigned to one of four experimental groups: normal animals with no treatment (Group 1, n=12); normal animals that underwent peritoneal implantation of porcine islets encapsulated in a double layer of agarose to form islet macrobeads (normal + islets; group 2, n=12); STZ treatment followed by daily exogenous insulin (STZ + insulin; group 3, n=18) and STZ treatment followed by the intraperitoneal implantation of porcine islet macrobeads (STZ + islets; group 4, n=14). At 215 days post-STZ induction, no renal proliferative lesions were observed in animals that did not receive STZ (group 1 and group 2) whereas adenoma incidences of 57% for group 3 and 34% for group 4 were observed. By terminal necropsy at day 351, the incidence and severity of renal proliferative lesions increased with tubular carcinoma observed in 67% of group 3 and 60% of group 4 animals. We conclude that the STZ-induced diabetic rat model is not suitable for long-term studies because of progressive renal tumorigenesis. Our experiments also demonstrate the safety and effectiveness of porcine islet macrobeads for the treatment of diabetes. PMID- 21633195 TI - Endomembrane Ca2+-ATPases play a significant role in virus-induced adaptation to oxidative stress. AB - Although the role of Ca2+ influx channels in oxidative stress signaling and cross tolerance in plants is well established, little is known about the role of active Ca2+ efflux systems in this process. In our recent paper, we reported Potato Virus X (PVX)-induced acquired resistance to oxidative stress in Nicotiana benthamiana and showed the critical role of plasma membrane Ca2+/H+ exchangers in this process. The current study continues this research. Using biochemical and electrophysiological approaches, we reveal that both endomembrane P2A and P2B Ca2+-ATPases play significant roles in adaptive responses to oxidative stress by removing excessive Ca2+ from the cytosol, and that their functional expression is significantly altered in PVX-inoculated plants. These findings highlight the crucial role of Ca2+ efflux systems in acquired tolerance to oxidative stress and open up prospects for practical applications in agriculture, after in-depth comprehension of the fundamental mechanisms involved in common responses to environmental factors at the genomic, cellular and organismal levels. PMID- 21633196 TI - Involvement of SGT1 in COR-mediated signal transduction pathway leading to disease symptom development. AB - Pseudomonas syringae pv. tomato DC3000 (Pst DC3000), that causes bacterial speck disease on tomato, produces a non-host-specific virulence effector, coronatine (COR). COR functions as a jasmonic acid (JA)-isoleucine mimic in planta and has multiple roles in the pathogenicity of Pst DC3000. One of the hallmarks of bacterial speck disease on tomato is the formation of necrotic lesions surrounded by chlorosis and COR is required for disease development. However, the molecular basis of COR-mediated disease symptom development including chlorosis and necrosis is still largely unknown. In our recent publication in New Phytologist, using virus-induced gene silencing (VIGS) based reverse genetics screen, we demonstrated that SGT1 (suppressor of G2 allele of skp1) is required for COR induced chlorosis in Nicotiana benthamiana. SGT1-silenced tomato leaves showed a complete loss of COR-induced chlorosis and reduced disease symptom development after the inoculation with Pst DC3000. Furthermore, Arabidopsis sgt1b mutant was less sensitive to COR-induced root growth inhibition and showed delayed Pst DC3000 disease symptoms. In this addendum, we discuss the possible contribution of SGT1 to COR-mediated signal transduction pathway leading to disease symptom development during Pst DC3000 pathogenesis in tomato and Arabidopsis. PMID- 21633197 TI - Unraveling the involvement of ABA in the water deficit-induced modulation of nitrogen metabolism in Medicago truncatula seedlings. AB - Effects of water deficit and/or abscisic acid (ABA) were investigated on early seedling growth of Medicago truncatula, and on glutamate metabolism under dark conditions. Water deficit (simulated by polyethylene glycol, PEG), ABA and their combination resulted in a reduction in growth rate of the embryo axis, and also in a synergistic increase of free amino acid (AA) content. However, the inhibition of water uptake retention induced by water deficit seemed to occur in an ABA-independent manner. Expression of several genes involved in glutamate metabolism was induced during water deficit, whereas ABA, in combination or not with PEG, repressed them. The only exception came from a gene encoding 1 pyrroline-5-carboxylate synthetase (P5CS) which appeared to be induced in an ABA dependent manner under water deficit. Our results demonstrate clearly the involvement of an ABA-dependent and an ABA-independent regulatory system, governing growth and glutamate metabolism under water deficit. PMID- 21633198 TI - An improved protocol for optical projection tomography imaging reveals lobular heterogeneities in pancreatic islet and beta-cell mass distribution. AB - Optical projection tomography (OPT) imaging is a powerful tool for three dimensional imaging of gene and protein distribution patterns in biomedical specimens. We have previously demonstrated the possibility, by this technique, to extract information of the spatial and quantitative distribution of the islets of Langerhans in the intact mouse pancreas. In order to further increase the sensitivity of OPT imaging for this type of assessment, we have developed a protocol implementing a computational statistical approach: contrast limited adaptive histogram equalization (CLAHE). We demonstrate that this protocol significantly increases the sensitivity of OPT imaging for islet detection, helps preserve islet morphology and diminish subjectivity in thresholding for tomographic reconstruction. When applied to studies of the pancreas from healthy C57BL/6 mice, our data reveal that, at least in this strain, the pancreas harbors substantially more islets than has previously been reported. Further, we provide evidence that the gastric, duodenal and splenic lobes of the pancreas display dramatic differences in total and relative islet and beta-cell mass distribution. This includes a 75% higher islet density in the gastric lobe as compared to the splenic lobe and a higher relative volume of insulin producing cells in the duodenal lobe as compared to the other lobes. Altogether, our data show that CLAHE substantially improves OPT based assessments of the islets of Langerhans and that lobular origin must be taken into careful consideration in quantitative and spatial assessments of the pancreas. PMID- 21633199 TI - Nutrient amino acids signal to mTOR via inositol polyphosphate multikinase. PMID- 21633200 TI - New players unveiled in early anther development. AB - Anther development is an important process for the successful sexual reproduction in plants. Whereas the regulation of the late stages of anther development is quite well described in A. thaliana, little is known about the regulation of the early stages of this process. Two novel groups of factors involved in these early stages have recently been described, namely ROXYs, members of the Glutaredoxin (GRX) family of small and ubiquitous oxidoreductases involved in various cellular and stress-related responses, and SBP-box Genes. ROXYs belong to the CC-type of GRXs with a CCXC active motif and are specific for higher plants. SBP-box genes encode for SQUAMOSA PROMOTER BINDING PROTEIN transcription factors, many of which are targeted by miR156 and miR157. Strikingly, both the enzymes and the transcription factors represent evolutionary conserved gene families and loss-of function of these genes exhibits similar anther phenotypes, e.g. arresting sporogenous cell formation and missing pollen sacs. This mini-review gives an overview of how these factors affect early anther development and discusses a possible relationship between these factors and other known early anther genes. PMID- 21633201 TI - The positive predictive value of guaiac faecal occult blood test in relation to the number of positive squares in two consecutive rounds of colorectal cancer screening. AB - The aim of this study was to define the positive predictive values of a positive guaiac faecal occult blood test according to the number of positive squares, in two consecutive rounds of colorectal cancer mass screening in a French region. A total of 4172 colonoscopies were analyzed. Sex, age, number of positive squares, and colonoscopic and histopathologic findings were studied. In the results obtained, 76.6% of positive tests were positive with one or two squares. The number of positive squares was not related to sex, age and rank of participation. The positive predictive value for cancers and adenomas increased significantly with age, sex (male) and number of positive squares from 6.6% (one to two squares) to 27.6% (five to six squares) and from 15.2% to 22.2%, respectively. Cancer was diagnosed 211 times (54.1%) and advanced neoplasia was diagnosed 696 times (65.3%) following positive tests with one to two squares. The TNM stage of cancer increased significantly with the number of positive squares: 85.8% of stages 0-1-2 for one to two positive squares and 66.3% for five to six positive squares (P<0.001). Multivariate analysis showed an increased risk of cancer and advanced neoplasia for male patients and aged persons. The number of positive squares significantly increased the risk of cancer (odds ratio=4.6 for five to six positive squares) and the risk of advanced neoplasia (odds ratio=2.9). Age, sex and number of positive squares were independent predictive factors of positive guaiac faecal occult blood test. The proportion of TNM stages 3-4 was significantly higher in those with five to six positive squares. Performing a complete colonoscopy in every individual having a positive test, especially aged men with a high number of positive squares, should be a priority in any screening programme. PMID- 21633202 TI - Chlorinated pesticides and cancer of the head and neck: a retrospective case series. AB - Cancer of the head and neck is a pervasive problem with recognized determinants including tobacco use, alcohol consumption, and earlier radiation exposure. Organochlorine pesticides (OCPs) have been shown to have carcinogenic potential in both animals and humans. OCPs have previously been widely used in the agricultural industry of rural Oklahoma. Seven patients from rural Oklahoma with head and neck cancer and without any of the usual risk factors were tested for the presence of OCPs in their adipose tissue. Clinical and toxicological data on each of these patients are presented for consideration. Results were compared with (i) levels from five individuals not experiencing cancer but who lived in the same area, and (ii) adipose tissue OCP levels in other population groups. Each of the seven patients tested had markedly elevated levels of some OCPs in their adipose tissue compared with the cohort of noncancer patients. Further research is required to confirm whether there is a causative link between OCP bioaccumulation and head and neck cancer as suggested by this case series. PMID- 21633203 TI - Renal cancer paradox: higher incidence but not higher mortality among African Americans. AB - To compare temporal trends in the incidence and mortality of renal cell cancer among blacks and whites for clues to etiologic differences. We examined trends in age-adjusted and age-specific Surveillance Epidemiology and End Results incidence and US mortality rates for renal cancer for 1973 through 2007, as well as nephrectomy rates from surgery codes for kidney cancer for 2000 through 2007. For nearly four decades, incidence rates for renal cell cancer have been rising more rapidly among blacks than whites, leading to a shift in excess from among whites to among blacks, almost entirely accounted for by an excess of localized disease. The incidence patterns are puzzling, as localized renal cell cancer is primarily detected incidentally by imaging, to which blacks have historically had less access. In contrast to the incidence patterns, there has been an unexpected convergence of renal cancer mortality rates, which have been virtually identical among blacks and whites since the early 1990 s. Nephrectomy rates, regardless of stage, were lower among blacks than among whites, despite almost identical cause specific survival rates in both races. The identical mortality patterns, combined with higher and more rapidly increasing incidence and lower rates of nephrectomies among blacks, suggest that renal cell cancer may tend to be a less aggressive tumor in blacks. This hypothesis is supported by the favorable stage distribution among blacks and their higher survival for distant and unstaged cancer. Further research into the enigmatic descriptive epidemiology and the biology and natural history of renal cell cancer may shed light on the etiology of this malignancy and its more frequent occurrence among black Americans. PMID- 21633204 TI - Moving data to nursing quality excellence. AB - Data must make their way to front line staff if performance improvement efforts are to be successful and sustainable. The steps of focus, display, distribute, and reward may be used as a process to ensure that data are delivered to and used by the front line staff. The focus on specific key measures, with subsequent consistent display and distribution, had a positive impact on the selected measures. Rewarding units with annual awards further encouraged staff to improve on important nurse-sensitive measures. PMID- 21633205 TI - Development and evaluation of a three-dimensional virtual hospital unit: VI-MED. PMID- 21633207 TI - HIMSS11 Nursing Informatics Symposium: informatics' role in bridging the quality chasm. PMID- 21633208 TI - Nursing informatics scope of practice expands, salaries increase. PMID- 21633209 TI - The health of American Indians. PMID- 21633210 TI - The health of American Indians. PMID- 21633211 TI - The coming of the blessing: A successful cross-cultural collaborative effort for American Indian/Alaska Native families. AB - This case study describes how the desire of an American Indian community in Wyoming grew into an American Indian/Alaska Native women's advisory committee, a culturally appropriate prenatal education booklet, and a national initiative for a nonprofit organization. The work began by bringing together women from 13 different nations and tribes, gaining the trust of the American Indian/Alaska Native women and working together to create an award winning booklet and a national initiative to support culturally appropriate prenatal education to every young American Indian/Alaska Native woman across this nation. Reservations from Wisconsin to Alaska have received this program for their young women. PMID- 21633212 TI - Engaging tribal leaders in an American Indian healthy eating project through modified talking circles. AB - Frequently used in the American Indian (AI) community and proven to be a valuable tool in health research, a talking circle is a method used by a group to discuss a topic in an egalitarian and nonconfrontational manner. Using community-based participatory research, a modified Talking Circle format was developed for engaging tribal leaders in an American Indian Healthy Eating Project in North Carolina. The culturally informed formative research approach enabled us to garner project support from 7 tribes, as well as insights on developing planning and policy strategies to improve access to healthy eating within each of the participating communities. PMID- 21633213 TI - The dynamic interplay between low socioeconomic status and diabetes for urban American Indians. AB - Socioeconomic status is a pervasive predictor of health and has a gradient effect on many diseases, such as diabetes. American Indians and Alaska Natives nationwide live in some of the most difficult socioeconomic conditions and have a higher prevalence of diabetes than any other major racial/ethnic group in the United States. This article contextualizes the nature of socioeconomic status in diabetes, diabetes management, and urban American Indians' lives by using a case study. Underscoring the economic dimensions in this manner can illuminate the complexities of life for urban American Indians with diabetes and guide diabetes initiatives for this population. PMID- 21633214 TI - Understanding parenting in Manitoba First nations: implications for program development. AB - This qualitative study introduced the "Manitoba First Nation Strengthening Families Maternal Child Health Pilot Project" program and evaluation methodologies. The study provided a knowledge base for programmers, evaluators, and communities to develop relevant health promotion, prevention, and intervention programming to assist in meeting health needs of pregnant women and young families. Sixty-five open-ended, semistructured interviews were completed in 13 communities. Data analysis was through grounded theory. Three major themes emerged from the data: interpersonal support and relationships; socioeconomic factors; and community initiatives. Complex structural, historical events compromise parenting; capacity and resilience are supported through informal and formal health and social supports. PMID- 21633215 TI - A women's shelter in a rural American indian community. AB - The author presents a case study and personal story about a group of community members initiating a women's shelter in a rural American Indian reservation community. Problems are identified and their resolutions described in organizing a community group, encountering community resistance, locating the shelter, raising funds, and developing procedures for operating the shelter. Lessons learned by the group are provided to illustrate possible solutions to problems that other community groups may encounter when seeking to open a women's shelter in a rural American Indian community. PMID- 21633216 TI - Family Wellness Warriors. AB - This article discusses the development and implementation of a culturally relevant resiliency model to address the physical, mental, emotional, and spiritual wellness needs of adults who have experienced harm. Common diseases in adulthood have been linked to abuse in childhood. To end abuse and improve health in Alaska, Alaska Native people are forming partnerships with health care organizations, government agencies, social service providers, and faith communities to disseminate Family Wellness Warriors Initiative strategies. Nearly 3000 people have participated in Family Wellness Warriors Initiative trainings. Through this healing work, families and communities are working together to change health disparities and restore hope. PMID- 21633217 TI - Fetal alcohol spectrum disorders: a Native American journey to prevention. AB - Fetal alcohol spectrum disorders, the most common preventable cause for mental retardation, is the result of prenatal alcohol exposure. There is no safe amount of alcohol during pregnancy. Native Americans have a higher risk of alcohol abuse than the general U.S. population. The fetal alcohol spectrum disorders prevalence rates for Native Americans range from 1.0 to 8.97 per 1000 births. Nurses and health care providers working in collaboration with tribal fetal alcohol spectrum disorders prevention specialists can greatly, and positively, impact the physical and mental health and well-being of children in Native American communities. PMID- 21633218 TI - Applying indigenous community-based participatory research principles to partnership development in health disparities research. AB - This case study of community and university research partnerships utilizes previously developed principles for conducting research in the context of Native American communities to consider how partners understand and apply the principles in developing community-based participatory research partnerships to reduce health disparities. The 7 partnership projects are coordinated through a National Institutes of Health-funded center and involve a variety of tribal members, including both health care professionals and lay persons and native and nonnative university researchers. This article provides detailed examples of how these principles are applied to the projects and discusses the overarching and interrelated emergent themes of sharing power and building trust. PMID- 21633219 TI - Promoting identities and mental health via cultural/community activities among racially/ethnically mixed urban American Indians. AB - This qualitative, focus group study explored the role of cultural/community activities in potentially promoting identities and mental health among urban American Indians who have mixed racial/ethnic heritages (e.g., African, Hispanic, and Caucasian) including those with mental illness in Philadelphia, Pennsylvania (n = 25). The study identified a variety of unique benefits of cultural/community activities including its role in nurturing identity, belongingness, spiritual wakening and renewal, and mental health. Overall, these activity-induced benefits helped urban American Indians gain a feeling of oneness to experience peace of mind and harmony within oneself and with other people and the environment/nature. PMID- 21633223 TI - Drinking from a fire hose-more than we can handle? PMID- 21633220 TI - Best practices guidelines for nurse practitioners regarding smoking cessation in American Indian and Alaskan Native youth. AB - The greatest prevalence of tobacco use in the United States occurs with the American Indian and Alaskan Native (AI/AN). A critical need exists for a culturally specific tobacco cessation option for AI/AN youth. The nurse practitioner is positioned to provide a culturally specific commercial tobacco cessation option by incorporating the transcultural nursing theory into the development of a decision tree to expand understanding of culturally appropriate best practices regarding screening and management of tobacco smoking cessation in AI/AN youth. Presented is the Nurse Practitioner Culturally Specific American Indian and Alaskan Native Youth Decision Tree for Smoking Cessation with supporting evidence-based best practices. PMID- 21633224 TI - Webcasting in home and hospice care services: virtual communication in home care. AB - The access to free live webcasting over home computers was much more available in 2007, when three military leaders from West Point, with the purpose of helping military personnel stay connected with their families when deployed, developed Ustream.tv. There are many types of Web-based video streaming applications. This article describes Ustream, a free and effective communication tool to virtually connect staff. There are many features in Ustream, but the most useful for home care and hospice service providers is its ability to broadcast sound and video to anyone with a broadband Internet connection, a chat room for users to interact during a presentation, and the ability to have a "co-host" or second person also broadcast simultaneously. Agencies that provide community-based services in the home will benefit from integration of Web-based video streaming into their communication strategy. PMID- 21633226 TI - Pressure ulcers at end of life: an overview for home care and hospice clinicians. PMID- 21633228 TI - Telehealth monitoring: a smart investment for home care patients with heart failure? AB - This telehealth project was positive overall. It was an agency goal to utilize telehealth to augment, not replace, SN visits while improving quality outcomes for the patient. The findings of this project supported that goal. PMID- 21633229 TI - Cost factors in implementing telemonitoring programs in rural home health agencies. AB - The purpose of this quantitative study was to examine costs and implementation factors associated with development of telemonitoring programs in eight rural home health agencies. although the telemonitoring group (n = 1,513) averaged fewer visits per episode than the control group (n = 1,573), cost analysis data, including labor, travel, and equipment costs, failed to support the hypothesis that decreased utilization of skilled nursing visits alone could offset the costs of the telemonitoring technology. this study indicates that agencies must achieve savings through improved outcome performance to offset telemonitoring expenses. PMID- 21633230 TI - Emergency management in action: surviving a flood. AB - On March 30, 2010, municipalities in southeastern Connecticut (CT) woke up to continued rainfall and were on the verge of the most significant flooding in over 50 years. This weather emergency would see the entire area affected in the subsequent weeks and months. What does this weather event have to do with home healthcare, you ask? Think of all the issues from a patient, staff, and community emergency management perspective and then think of how to deliver and ensure quality home healthcare services. Once again, home healthcare nurses (HHNs) were placed in the forefront of a community in need in such an instance. PMID- 21633231 TI - Remote telemonitoring technology will expand, but progress depends on more research and federal support. PMID- 21633234 TI - Management of cataracts in patients with glaucoma. AB - The management of the glaucoma patient who has visually significant cataracts is a practical but complex topic. As glaucoma is a chronic, potentially progressive disease that can lead to irreversible blindness, ophthalmologists should develop a treatment approach with emphasis based on the severity of glaucoma rather than on cataract alone. Trabeculectomy remains an effective surgical choice, especially in glaucoma patients with advanced disease who require a low and steady IOP. In planning for cataract surgery, surgeons should be mindful of the alterations of astigmatism in terms of power and axis, axial length fluctuation as a result of trabeculectomy, and the relative position of the IOL after surgery. As glaucoma is a potentially progressive disease, surgeons who treat patients with coexistent glaucoma and cataracts must consider that future glaucoma surgery may be necessary when planning for the cataract surgery. A complete discussion of alternatives would go beyond the limited scope of this study. Regardless of the procedures used, the surgeon should consider the secondary effects of both glaucoma surgery and cataract surgery and their impact on each other when developing an individualized treatment plan. PMID- 21633235 TI - Ocular perfusion pressure and glaucoma. PMID- 21633236 TI - Medical and surgical treatment of neovascular glaucoma. PMID- 21633237 TI - Genetics of adult glaucoma. PMID- 21633238 TI - Myopia and glaucoma. PMID- 21633239 TI - Ab interno trabeculotomy with the trabectome system: what does the data tell us? PMID- 21633240 TI - Ex-PRESS glaucoma filtration device-surgical technique and outcomes. PMID- 21633241 TI - New methods and modifications of glaucoma filtration surgery. PMID- 21633242 TI - Genetics of the pediatric glaucomas. PMID- 21633243 TI - Aqueous humor dynamics and implications for clinical practice. PMID- 21633244 TI - Interpretation and misinterpretation of results from the tube versus trabeculectomy study. PMID- 21633245 TI - Update on aqueous shunts in glaucoma. PMID- 21633246 TI - Laser trabeculoplasty in the treatment of open-angle glaucoma. PMID- 21633248 TI - Atrial fibrillation and atrial fibrosis. AB - Atrial fibrillation (AF) is the most common arrhythmia in humans. It affects 5% of the population older than age 65 years and is projected to rise as the population ages. Experimental data from animal models of AF show that AF is associated with progressive structural and electrical remodeling of the atria. Atrial fibrosis alters atrial electrical conduction and excitability and provides a substrate for AF maintenance. However, whether fibrosis is causally related to AF or an epiphenomenon and the precise mechanisms underlying atrial fibrosis remain unclear. A variety of signaling systems involving angiotensin II and related mediators are centrally involved in atrial fibrosis. This article reviews the role that atrial fibrosis plays in AF, the mechanisms of atrial fibrosis, and emerging therapeutic approaches to AF aimed at attenuating atrial fibrosis. PMID- 21633250 TI - EEG patterns and imaging correlations in encephalopathy: encephalopathy part II. AB - The EEG patterns seen with encephalopathies can be correlated to cerebral imaging findings including head computerized tomography and MRI. Background slowing without slow-wave intrusion is seen with acute and chronic cortical impairments that spare subcortical white matter. Subcortical/white matter structural abnormalities or hydrocephalus may produce projected slow-wave activity, while clinical entities involving both cortical and subcortical regions (diffuse cerebral abnormalities) engender both background slowing and slow-wave activity. Triphasic waves are seen with hepatic and renal insufficiency or medication toxicities (e.g., lithium, baclofen) in the absence of a significant cerebral imaging abnormality, Conversely, subcortical/white matter abnormalities may facilitate the appearance of triphasic waves without significant hepatic, renal, or toxic comorbidities. More specific syndromes, such as Jakob-Creutzfeldt disease, autoimmune limbic encephalitis, autoimmune corticosteroid-responsive encephalopathy with thyroid autoimmunity, sepsis-associated encephalopathy, and acute disseminated encephalomyelitis, have imaging/EEG changes that are variable but which may include slowing and epileptiform activity. This overview highlighting EEG-imaging correlations may help the treating physician in the diagnosis, and hence the appropriate treatment, of patients with encephalopathy. PMID- 21633249 TI - Antiadrenergic and hemodynamic effects of ranolazine in conscious dogs. AB - Effects of ranolazine alone and in the presence of phenylephrine (PE) or isoproterenol (ISO) on hemodynamics, coronary blood flow and heart rate (HR) in the absence and presence of hexamethonium (a ganglionic blocker) were studied in conscious dogs. Ranolazine (0.4, 1.2, 3.6, and 6 mg/kg, intravenous) alone caused transient (<1 minute) and reversible hemodynamic changes. PE (0.3-10 MUg/kg) caused a dose-dependent increase in blood pressure and decrease in HR. ISO (0.01 0.3 MUg/kg) caused a dose-dependent decrease in blood pressure and an increase in HR. Ranolazine at high (11-13 mM), but not at moderate (4-5 mM) concentrations partially attenuated changes in mean arterial blood pressure and HR caused by either PE or ISO in normal conscious dogs. However, in dogs treated with hexamethonium (20 mg/kg) to cause autonomic blockade, ranolazine (both 4-5 and 11 13 MUM) significantly attenuated both the PE- and ISO-induced changes in mean arterial blood pressure. The results suggest that a potential antiadrenergic effect of ranolazine was masked by autonomic control mechanisms in conscious dogs but could be observed when these mechanisms were inhibited (eg, in the hexamethonium-treated dog). Ranolazine, at plasma concentrations <10 MUM and in conscious dogs with intact autonomic regulation, had minimal antiadrenergic (alpha and beta) effects. PMID- 21633251 TI - Artifact and recording concepts in EEG. AB - Artifact is present when electrical potentials that are not brain derived are recorded on the EEG and is commonly encountered during interpretation. Many artifacts obscure the tracing, while others reflect physiologic functions that are crucial for routine visual analysis. Both physiologic and nonphysiologic sources of artifact may act as source of confusion with abnormality and lead to misinterpretation. Identifying the mismatch between potentials that are generated by the brain from activity that does not conform to a realistic head model is the foundation for recognizing artifact. Electroencephalographers are challenged with the task of correct interpretations among the many artifacts that could potentially be misleading, resulting in an incorrect diagnosis and treatment that may adversely impact patient care. Despite advances in digital EEG, artifact identification, recognition, and elimination are essential for correct interpretation of the EEG. The authors discuss recording concepts for interpreting EEG that contains artifact. PMID- 21633252 TI - Artifact: recording EEG in special care units. AB - Artifacts may be obtained during routine recording but are more common in special care units (SCUs) outside of the EEG laboratory, where complex electrical currents are present that create a "hostile" environment. Special care units include the epilepsy monitoring unit, neurologic intensive care unit, and operating room, where artifact is present in virtually every recording, increasing with prolonged use. Nonepileptic attacks treated as epileptic seizures have been incorrectly diagnosed and treated due to a misinterpreted EEG. The recent emergence of continuous EEG as a neurophysiologic surrogate for brain function in the neurologic intensive care unit and operating room has also brought a greater amount and new types of EEG artifact. The artifacts encountered in special care units during continuous EEG are becoming more complex and may have adverse therapeutic implications. Our knowledge of artifact needs to parallel our growth in technology to avoid the pitfalls that may be incurred during visual analysis of the EEG. PMID- 21633253 TI - Rethinking clinical language mapping approaches: discordant receptive and expressive hemispheric language dominance in epilepsy surgery candidates. AB - Neuroimaging studies have shed light on cortical language organization, with findings implicating the left and right temporal lobes in speech processing converging to a left-dominant pattern. Findings highlight the fact that the state of theoretical language knowledge is ahead of current clinical language mapping methods, motivating a rethinking of these approaches. The authors used magnetoencephalography and multiple tasks in seven candidates for resective epilepsy surgery to investigate language organization. The authors scanned 12 control subjects to investigate the time course of bilateral receptive speech processes. Laterality indices were calculated for left and right hemisphere late fields ~150 to 400 milliseconds. The authors report that (1) in healthy adults, speech processes activated superior temporal regions bilaterally converging to a left-dominant pattern, (2) in four of six patients, this was reversed, with bilateral processing converging to a right-dominant pattern, and (3) in three of four of these patients, receptive and expressive language processes were laterally discordant. Results provide evidence that receptive and expressive language may have divergent hemispheric dominance. Right-sided receptive language dominance in epilepsy patients emphasizes the need to assess both receptive and expressive language. Findings indicate that it is critical to use multiple tasks tapping separable aspects of language function to provide sensitive and specific estimates of language localization in surgical patients. PMID- 21633254 TI - Covariation of EEG synchronization and emotional state as modified by anxiolytics. AB - The relationships between subjective estimation of emotional state and synchronization patterns in cortical emotional systems were investigated. The emotional state varied between groups using diazepam, buspirone, and placebo. The University of Wales Institute of Science and Technology Mood Adjective Checklist was used for the assessment of emotional state in the drug condition, yielding three estimates of emotional state: Energetic Arousal, Tension Arousal, and Hedonic Tone. These measures were correlated with the Synchronization Likelihood index of the resting EEG. Increased affective valence and arousal were related to an increased level of synchronization between frontal and right temporoparietal emotional areas. Two identified centers of synchronization, localized in the temporal and centroparietal regions, appeared to be functionally distinct. Stable relationships between subjective emotional state measures and cortical EEG synchronization patterns were confirmed, especially for the valence and energetic arousal estimation. A higher synchronization is associated with increased emotional valence and arousal, and this can thus be seen as a neural correlate of emotional experiences. PMID- 21633255 TI - Corticospinal descending direct wave elicited by subcortical stimulation. AB - Recent studies have indicated the importance of subcortical mapping of the corticospinal tract (CT) during tumor resection close to the primary motor area. It is substantial evidence that the corticospinal descending direct wave (D-wave) can be used as a guide for mapping of the primary motor cortex (M1) and for monitoring of the CT functional integrity. In the present study, the authors investigated the feasibility of D-wave recordings after subcortical stimulation. The authors examined 14 patients with brain tumors close to the M1 and/or CT, who exhibited no obvious motor deficit before surgery. Subcortical white matter was electrically stimulated in monopolar or bipolar fashion by recording the descending wave (D-wave) from the spinal epidural space using a catheter-type electrode. Subcortical D-wave was more clearly recorded after monopolar stimulation than after bipolar stimulation. The features of the subcortical D wave, including its waveform, conduction velocity, and latency, were nearly identical to those of the corticospinal D-wave recorded after M1 stimulation. Subcortical D-wave amplitude was prone to change depending on the distance from the stimulation points to the CT. Changes in parameters of subcortical D-wave may provide valuable information to prevent postoperative motor deficit. Further studies are required to clarify the relationship between the distance from the stimulating point to the CT and the amplitude of the subcortically elicited D wave. PMID- 21633256 TI - Auditory evoked potential P50 as a predictor of neurologic outcome in resuscitated cardiac arrest patients. AB - In general, a prediction of neurologic outcome with respect to the resuscitated cardiac arrest patients has been performed by the auditory brainstem response and somatic evoked potential. The auditory brainstem response and somatic evoked potential are known as the predictors that correspond to neurologically poor outcome. None of the methods have been established to access neurologically good outcome. Because the hippocampal CA3 pyramidal cells have been widely used for pathophysiologic analyses concerning the hypoxic-ischemic encephalopathy and also the source of P50 components of the auditory evoked potential has been considered to be the hippocampal CA3 pyramidal cells, the authors assume that it might be possible that neurologic outcome in resuscitated cardiac arrest patients would be predicted by evaluating the P50 components. The purpose was to examine the P50 as a predictor of neurologic outcome in resuscitated cardiac arrest patients at the early stage from the onset. The P50 components of the auditory evoked potential are recorded in a conditioning-testing paradigm, that is, EEG responses to a pair of auditory stimuli with 500-millisecond interclick interval. In this study, subjects are 10 out-of-hospital cardiac arrest patients, 8 men and 2 women with a mean age of 54.8 years, who were admitted to the intensive care unit after the return of spontaneous circulation, with the presence of both the auditory brainstem response wave V and the somatic evoked potential wave N20 between the period from June 2008 to July 2009. It was found that the presence of the P50 at the early stage from the onset (days 5 +/- 1.20) indicates good neurologic outcome, while the absence of the P50 implies poor prognosis. As to the auditory sensory gating of the P50, almost no reduction response to the second stimulus was observed. As a consequence, the evaluation of the P50 in resuscitated cardiac arrest patients would have a possibility to predict neurologically good outcome. PMID- 21633257 TI - Physiologic pseudoseizures: an EEG case report of mistake in identity. AB - The misdiagnosis of epilepsy may occur from the misinterpretation of a routine scalp EEG. Typically, interictal epileptiform discharges are misidentified on EEGs because of the overinterpretation of normal variants or variations in normal electrocerebral activity. Most reports of misinterpretation have arisen from patients diagnosed with psychogenic nonepileptic attacks using in-patient video EEG monitoring. However, because seizures are rarely captured in the routine performance of EEG in the outpatient setting, ictal overinterpretation is much less likely to occur. However, some physiologic conditions may also occur with paroxysmal hypersynchronous patterns on EEG and lead to a misdiagnosis of epilepsy. We report an adult woman with untreated obstructive sleep apnea and major depressive disorder that was self-referred for the evaluation of seizures. Brief episodes of delayed responsiveness associated with prolonged reading demonstrated the "generalized paroxysmal pattern" that was misinterpreted as a seizure during routine EEG at an outside institution. Subsequently, in-patient video-EEG monitoring reproduced multiple nonepileptic physiologic paroxysmal hypersynchronous arousals after prolonged reading. Treatment of the underlying sleep apnea, psychologic support, and limited prolonged reading lead to remission of her nonepileptic "spells" and successful discontinuation of antiepileptic drugs. PMID- 21633258 TI - What is the diagnostic value of repeating a nondiagnostic video-EEG study? AB - Repeat video-EEG (VEEG) may increase diagnostic yield, but the test is resource intensive, time-consuming, and expensive and poses some potential risks to patients. It is also relatively common to monitor a patient for several days without capturing any clinical events. The purpose of this study was to determine the diagnostic value of repeat admissions for VEEG and to determine if the commonly available clinical information could predict the diagnostic outcome, "diagnostic" or "nondiagnostic," of a repeat study. A study was deemed diagnostic if the admission resulted in a definitive diagnosis of the patient's typical events. The authors retrospectively reviewed the charts of 3,727 patients completing scalp VEEG at the University of Alabama at Birmingham Epilepsy Center from 2002 to 2009. Minors, mentally retarded patients, and patients readmitted for surgical procedures or presurgical localization were excluded. Single and multiple regressions were used to determine if any of the parameters could predict the diagnostic outcome of a repeat VEEG study. Only younger age was independently predictive of a diagnostic readmission (P < 0.05), while longer total duration of monitoring was suggestive (P = 0.07). A repeat VEEG study was useful in 55.2% of patients, most of whom were diagnosed after only 1 additional admission. In the patient population studied, 82.4% were diagnosed on the first admission (2,622 of 3,183), 52.9% on the second (46 of 87), and 40% on the third (2 of 5). Repeat VEEG admissions are useful, and clinical expertise may be the best tool for planning potential readmissions. PMID- 21633259 TI - Increased sleep spindle activity in patients with Costello syndrome (HRAS gene mutation). AB - Costello syndrome is a congenital disorder because of HRAS gene mutation, frequently associated with neurologic impairment and sleep disorders. The aims of the study were to evaluate the sleep EEG, and particularly the sleep spindles, in a population of patients with Costello syndrome and to compare them with those characterizing unaffected subjects. Eleven subjects (5 men and 6 women) with Costello syndrome were included in the study; age ranged between 18 months and 31 years (mean, 9.6 +/- 9.4 years). The diagnosis was posed on the basis of established clinical criteria and confirmed molecularly. Sleep EEG was studied by means of full-night, laboratory-based video-polysomnography, performed overnight, during hospitalization. Sleep activity was quantified by means of power spectral analysis. Patients heterozygous for an HRAS mutation exhibited increased EEG power in 12- to 15-Hz activity band compared with age-matched control subjects. In conclusion, the authors observed a consistent increase in the amplitude of cortical sleep spindles in all our subjects with an HRAS mutation. These "giant" spindles were not associated with any evidence of structural damage of the cortex or the thalami and should be considered as phenotypic feature of sleep EEG activity in Costello syndrome because of HRAS mutation. PMID- 21633260 TI - Effects of 1-Hz repetitive transcranial magnetic stimulation on long-latency reflexes and cortical relay time. AB - Long-latency reflexes (LLRs) of hand muscles include a transcortical component. Cortical relay time estimated by the subtraction of motor and somatosensory evoked potentials from LLR reflects the physiology of the central neural pathway of LLR. It is believed that 1-Hz repetitive transcranial magnetic stimulation (rTMS) of the primary motor cortex can decrease cortical excitability for approximately 15 minutes at intracortical level. The aim of the study was to analyze LLR and cortical relay time before and after 1-Hz rTMS. Long-latency reflex and H reflex obtained from the thenar muscles by electrical stimulation of the median nerve of 16 healthy subjects. Additionally, motor evoked potentials and somatosensory evoked potentials were also recorded. Cortical relay time was calculated by the subtraction of motor evoked potential and somatosensory evoked potential latencies from LLR. These electrophysiologic recordings were performed before and after 15 minutes of 1-Hz rTMS over the motor area for the thenar muscles in the primary motor cortex. The amplitudes of LLR and motor evoked potential were significantly decreased after rTMS, but the H reflex of the thenar muscle and somatosensory evoked potentials were unchanged. The major finding of our study was a shortened duration of cortical relay time after rTMS. In conclusion, our findings suggest that the LLR of the thenar muscles has a transcortical pathway and cortical relay time that can give some information about the physiology of the intracortical pathway of LLR. PMID- 21633261 TI - Electrophysiologic changes in patients with untreated primary hypothyroidism. AB - Thyroid diseases may cause signs and symptoms of neuromuscular dysfunction. Hypothyroidism has been associated with the clinical features of proximal muscle weakness, mononeuropathy, and sensorimotor polyneuropathy. This study aimed at evaluating the electrophysiologic findings in patients with untreated spontaneous hypothyroidism and comparing them with a healthy control group. In a case-control cross-sectional study, 40 patients with definite diagnosis of clinical hypothyroidism and 40 healthy control subjects were evaluated by electromyography and nerve conduction studies in the specialized clinic of the Tabriz University of Medical Sciences during an 18-month period. Seven male and 33 female patients with a mean age of 39.5 +/- 11.8 years were enrolled. In this group, there were 12 cases (30%) with clinical muscle weakness, with severity of approximately 4/5, 18 cases (45%) with decreased or absent deep tendon reflexes, 6 cases (15%) with neuropathy, including 4 sensory and 2 sensorimotor, of which 5 cases were mild and 1 case was moderate, 3 cases (7.5%) with myopathy, and 13 cases (32.5%) with carpal tunnel syndrome, which was mild in 7, moderate in 10, and severe in 2 hands. Patients with neuropathy were significantly older than those without neuropathy (P = 0.001). There was no significant relation between gender, duration of the disease, serum TSH level, and the presence of clinical muscle weakness with the occurrence of neuropathy or myopathy. Female gender, increasing age, duration of the disease, and the frequency of clinical weakness were, however, significantly related to the presence of carpal tunnel syndrome (P < 0.05). In conclusion, in patients with untreated primary hypothyroidism, majority had the carpal tunnel syndrome. Mild neuropathy mainly of sensory type and myopathy were uncommon and rare findings, respectively. Early treatment would hinder the progression of mentioned abnormalities and minimize their occurrence. PMID- 21633262 TI - Association between neuropathic pain and A-waves in leprosy patients with type 1 and 2 reactions. AB - Neuropathic pain (NP) is a well-recognized feature of leprosy neuropathy. However, the diagnosis of NP is difficult using only clinical criteria. In the study reported here, by means of conventional nerve conduction studies, the authors sought for an association between long-latency responses and NP complaints in leprosy patients with type 1 and 2 reactions. Of the 27 ulnar nerves of leprosy patients, 18 with type 1 reaction (T1R) and 9 with type 2 reaction (T2R) were followed-up for 6 months before and after steroid treatment. Clinical characteristics of pain complaints and clinical function were assessed, as well as the presence of F- and A-waves of the ulnar nerve using nerve conduction studies. The clinical and the neurophysiologic findings were compared to note positive concordances (presence of NP and A-waves together) and negative concordances (absence of NP and A-waves together) before and after treatment. Both reactions presented a high frequency of A-waves (61.1% in T1R and 66.7% in T2R, P < 0.05) and prolonged F-waves (69.4% in T1R and 65.8% in T2R, P = 0.4). No concordances were seen between pain complaints and F-waves. However, significant concordances between NP and A-waves were observed, although restricted to the T2R group (chi(2) = 5.65, P = 0.04). After treatment, there was a significant reduction in pain complaints, as well as the presence of F- and A-waves in both groups (P < 0.05 for all comparisons). In conclusion, the presence of A-waves correlates well with pain complaints of neuropathic characteristics in leprosy patients, especially in those with type 2 reaction. Probably, such response shares similar mechanisms with the small-fiber dysfunction seen in these patients with NP, such as demyelination, intraneural edema, and axonal sprouting. Further studies using specific tools for small-fiber assessment are warranted to confirm our findings. PMID- 21633264 TI - Prognostic value of the EEG in anoxic encephalopathy. PMID- 21633266 TI - Plastic surgery on the rise. PMID- 21633267 TI - Nurses: needed and valued. PMID- 21633268 TI - Letter from the president of PSNCB. PMID- 21633269 TI - Interruptions and distractions during anesthesia induction: a pilot study. AB - Usually, the circulating registered nurse assists with anesthesia induction, but other personnel tend to interrupt the process to ask questions that are unrelated to the induction process. Interruptions and distractions can lead to loss of focus and result in medication errors if a certified registered nurse anesthetist (CRNA) swaps syringes during anesthesia induction. In this study, 8 CRNAs and cycles of anesthesia induction were observed, and the number of interruptions and distractions recorded. Results showed that most were from other personnel (M = 3 +/- 0.53), conversation (M = 3 +/- 1.19), and noise (M = 1.5 +/- 1.3). With an average of 7.5 total interruptions per 9 min, these results indicate that CRNAs may experience 68 interruptions and distractions per hour. In another setting, this number is likely to be different. We have chosen to publish this provocative article to stimulate other similar studies of interruptions and distractions in the surgical setting. More research may show whether silence during induction is upheld as a safety measure, and what might be solutions to distractions and interruptions. PMID- 21633270 TI - Specialty nurse certification effects patient outcomes. AB - This article reports on studies that have shown specialty nurses with certification contribute to quality patient outcomes and satisfaction. In addition, nurses who have achieved their specialty certification have perceived intrinsic value, empowerment and heighten collaboration with the health care team. PMID- 21633271 TI - Transformation in Haiti: finding purpose while responding to disaster. AB - This is a human interest story and update on the current state of healthcare and earthquake recovery in Haiti. This is drawn from experiences the author has gained through disaster response after the 7.0 earthquake and in returning to Haiti as a medical volunteer in October 2010. What has been accomplished in the last year since the earthquake and the challenges that lay ahead for the Haitian people are addressed through personal experiences with patients, American volunteers, and local healthcare staff. The experience of being a medical volunteer in Port au Prince, Haiti, is portrayed through the surrounding environment, interactions with patients and fellow volunteers, and the resulting emotional and psychological impact. This piece culminates with the impression such volunteer work has left on the author and how it has been life-changing. PMID- 21633272 TI - Navigating new technologies in negative pressure wound therapy. AB - Plastic surgeons and their support staff are tasked with proficient management of a wide variety of complex wounds. Since its introduction, negative pressure wound therapy (NPWT) has increasingly been used within the plastic surgery specialty to improve and simplify wound management. Increased usage of the therapy has prompted the development of a myriad of new NPWT systems. While an expanded product selection allows greater choice to maximize patient outcomes, sound decision-making also requires a clear understanding of the characteristics of various NPWT systems and applications. Wound-specific NPWT systems of varying size are available for low- to moderate-severity wounds, clean closed incisions, and acute abdominal wounds. Wound size and severity, amount of exudate, and patient mobility issues have become important considerations when choosing an NPWT device. The purpose of this article is to familiarize the reader with the latest sophistications in NPWT systems to guide decision making and usage. PMID- 21633274 TI - Sinus bradycardia and sinus tachycardia. PMID- 21633276 TI - Wound photography: considerations and recommendations. PMID- 21633277 TI - Will the current economic environment, healthcare reform, and potential federal and state budget cuts create the "perfect storm" for insurance coverage of breast reduction surgeries? PMID- 21633278 TI - Effective leadership. PMID- 21633281 TI - The Madrid resolution on organ donation and transplantation: national responsibility in meeting the needs of patients, guided by the WHO principles. PMID- 21633282 TI - Executive summary. PMID- 21633280 TI - Third WHO Global Consultation on Organ Donation and Transplantation: striving to achieve self-sufficiency, March 23-25, 2010, Madrid, Spain. PMID- 21633283 TI - Report of the Madrid Consultation: Part 1: European and universal challenges in organ donation and transplantation, searching for global solutions. PMID- 21633284 TI - Report of the Madrid Consultation: Part 2: Reports from the working groups. PMID- 21633285 TI - HIV-1 drug resistance at antiretroviral treatment initiation in children previously exposed to single-dose nevirapine. AB - OBJECTIVE: To describe the prevalence of HIV-1 drug resistance mutations at the time of treatment initiation in a large cohort of HIV-infected children previously exposed to single-dose nevirapine (sdNVP) for prevention of transmission. DESIGN: Drug resistance mutations were measured pretreatment in 255 infants and young children under 2 years of age in South Africa exposed to sdNVP and initiating ritonavir-boosted lopinavir-based therapy. Those who achieved viral suppression were randomized to either continue the primary regimen or to switch to a nevirapine-based regimen. Pretreatment samples were tested using population sequencing and real time allele-specific PCR (AS-PCR) to detect Y181C and K103N minority variants. Those with confirmed viremia more than 1000 copies/ml by 52 weeks postrandomization in the switch group were defined as having viral failure. RESULTS: Nonnucleoside reverse transcriptase inhibitor (NNRTI) mutations, predominantly Y181C, were detected by either method in 62% of infants less than 6 months of age, in 39% of children 6-12 months of age, 22% 12 18 months, and 16% 18-24 months (P = <0.0001). NNRTI mutations detected by genotyping, but not K103N or Y181C mutations detected only by AS-PCR, were associated with viral failure in the switch group. CONCLUSION: The prevalence of mutations known to compromise primary NNRTI-based therapy is high in sdNVP exposed children, supporting current guidelines recommending use of protease inhibitor-based regimens for young children. Standard genotyping is adequate to identify children who could benefit from switching to NNRTI-based therapy. PMID- 21633286 TI - Synergy against drug-resistant HIV-1 with the microbicide antiretrovirals, dapivirine and tenofovir, in combination. AB - OBJECTIVE: To evaluate the candidate antiretroviral microbicide compounds, dapivirine (DAP) and tenofovir (TFV), alone and in combination against the transmission of wild-type and nonnucleoside reverse transcriptase inhibitor (NNRTI)-resistant HIV-1 from different subtypes. DESIGN AND METHODS: We determined single-drug efficacy of the RTIs, DAP and TFV, against subtype B and non-B wild-type and NNRTI-resistant HIV-1 in vitro. To assess breadth of activity, compounds were tested alone and in combination against wild-type and NNRTI-resistant subtype C primary HIV-1 isolates and complimentary clonal HIV-1 from subtypes B, C and CRF02_AG to control for viral variation. Early infection was quantified by counting light units emitted from TZM-bl cells less than 48-h postinfection. Combination ratios were based on drug inhibitory concentrations (IC(50)s) and combined effects were determined by calculating combination indices. RESULTS: Both candidate microbicide antiretrovirals demonstrated potent anti-NNRTI-resistant HIV-1 activity in vitro, albeit the combination protected better than the single-drug treatments. Of particular interest, the DAP with TFV combination exhibited synergy (50% combination index, CI(50) = 0.567) against subtype C NNRTI-resistant HIV-1, whereas additivity (CI(50) = 0.987) was observed against the wild-type counterpart from the same patient. The effect was not compounded by the presence of subdominant viral fractions, as experiments using complimentary clonal subtype C wild-type (CI(50) = 0.968) and NNRTI-resistant (CI(50) = 0.672) HIV-1, in lieu of the patient quasispecies, gave similar results. CONCLUSION: This study supports the notion that antiretroviral drug combinations may retain antiviral activity against some drug-resistant HIV-1 despite subtype classification and quasispecies diversity. PMID- 21633287 TI - The role of coinfections in HIV epidemic trajectory and positive prevention: a systematic review and meta-analysis. AB - OBJECTIVES: Recurrent or persistent coinfections may increase HIV viral load and, consequently, risk of HIV transmission, thus increasing HIV incidence. We evaluated the association between malaria, herpes simplex virus type 2 (HSV-2) and tuberculosis (TB) coinfections and their treatment on HIV viral load. DESIGN: Systematic review and meta-analysis of the association of malaria, HSV-2 and TB coinfections and their treatment with HIV viral load. METHODS: PubMed and Embase databases were searched to 10 February 2010 for studies in adults that reported HIV plasma and/or genital viral load by coinfection status or treatment. Meta analyses were conducted using random-effects models. RESULTS: Forty-five eligible articles were identified (six malaria, 20 HSV-2 and 19 TB). There was strong evidence of increased HIV viral load with acute malaria [0.67 log(10) copies/ml, 95% confidence interval (CI) 0.15-1.19] and decreased viral load following treatment (-0.37 log(10) copies/ml, 95% CI -0.70 to -0.04). Similarly, HSV-2 infection was associated with increased HIV viral load (0.18 log(10) copies/ml, 95% CI 0.01-0.34), which decreased with HSV suppressive therapy (-0.28 log(10) copies/ml, 95% CI -0.36 to -0.19). Active TB was associated with increased HIV viral load (0.40 log(10) copies/ml, 95% CI 0.13-0.67), but there was no association between TB treatment and viral load reduction (log(10) copies/ml 0.02, 95% CI -0.19 to 0.15). CONCLUSION: Coinfections may increase HIV viral load in populations where they are prevalent, thereby facilitating HIV transmission. These effects may be reversed with treatment. However, to limit HIV trajectory and optimize positive prevention for HIV-infected individuals pre-antiretroviral therapy, we must better understand the mechanisms responsible for augmented viral load and the magnitude of viral load reduction required, and retune treatment regimens accordingly. PMID- 21633288 TI - Elevated triglycerides and risk of myocardial infarction in HIV-positive persons. AB - OBJECTIVES: To explore the relationship between elevated triglyceride levels and the risk of myocardial infarction (MI) in HIV-positive persons after adjustment for total cholesterol (TC), high-density lipoprotein-cholesterol (HDL-C) and nonlipid risk factors. BACKGROUND: Although elevated triglyceride levels are commonly noted in HIV-positive individuals, it is unclear whether they represent an independent risk factor for MI. METHODS: The incidence of MI during follow-up was stratified according to the latest triglyceride level. Multivariable Poisson regression models were used to describe the independent association between the latest triglyceride level and MI risk after adjusting for TC and HDL-C, nonlipids cardiovascular disease (CVD) risk factors, HIV and treatment-related factors. RESULTS: The 33,308 persons included in the study from 1999 to 2008 experienced 580 MIs over 178,835 person-years. Unadjusted, the risk of MI increased by 67% [relative risk (RR) 1.67, 95% confidence interval 1.54-1.80] per doubling in triglyceride level. After adjustment for the latest TC and HDL-C level, the RR dropped to 1.33 (95% confidence interval 1.21-1.45); this effect was further attenuated by other CVD risk factors and the RR was reduced to 1.17 (95% confidence interval 1.06-1.29). In models that additionally adjusted for HIV and treatment factors, the risk was further diminished, although remained significant (RR 1.11, 95% confidence interval 1.01-1.23). CONCLUSION: Higher triglyceride levels were marginally independently associated with an increased risk of MI in HIV-positive persons, although the extent of reduction in RR after taking account of latest TC, latest HDL-C and other confounders suggests that any independent effect is small. PMID- 21633289 TI - Concurrent chemotherapy and intensity-modulated radiotherapy for organ preservation of locoregionally advanced oral cavity cancer. AB - OBJECTIVES: To report outcomes of oral cavity cancer patients treated with concurrent chemotherapy and intensity-modulated radiotherapy (chemoIMRT). METHODS: Between 2001 and 2004, 21 patients with oral cavity squamous cell carcinoma underwent definitive chemoIMRT. Sites included were oral tongue (n = 9), floor of mouth (n = 6), buccal mucosa (n = 3), retromolar trigone (n = 2), and hard palate (n = 1). Most had stage III-IV disease (n = 20). The most common regimen was 5 days infusional 5-fluorouracil (600 mg/m(2)/d * 5 days), hydroxyurea (500 mg, PO BID), and 1.5 Gy twice-daily irradiation to 72 to 75 Gy. RESULTS: The median follow-up for surviving patients was 60 months. Treatment failure occurred as follows: local-1, regional-1, and distant metastases-2. The 2 and 5-year estimates of locoregional progression-free survival, disease-free survival, and overall survival were 90% and 90%, 71% and 71%, and 76% and 76%, respectively. Late complications included osteoradionecrosis (3 patients, 14%). CONCLUSIONS: Concurrent chemoIMRT results in promising locoregional control for oral cavity squamous cell carcinomas with acceptable toxicity. PMID- 21633290 TI - Curcumin and resveratrol synergistically stimulate p21 and regulate cox-2 by maintaining adequate zinc levels during lung carcinogenesis. AB - This study explored the efficacy of curcumin and resveratrol in maintaining adequate zinc levels to regulate p21 and cyclooxygenase-2 (cox-2) during benzo[a]pyrene (BP)-induced lung carcinogenesis. The mice were segregated into five groups, which included normal control, BP treated, BP plus curcumin treated, BP plus resveratrol treated, and BP plus curcumin plus resveratrol-treated groups. BP treatment resulted in a significant decrease in the zinc levels and protein expression of p21. On the contrary, the enzyme activity of cox-2 showed a significant increase in the BP-treated mice. Interestingly, combined supplementation of curcumin and resveratrol to BP-treated mice resulted in an appreciable improvement in the zinc levels and protein expression of p21. In contrast, synergistic supplementation with phytochemicals resulted in a significant decrease in the enzyme activities of cox-2 in BP-treated mice. This study, therefore, concludes that combined treatment with curcumin and resveratrol maintains adequate zinc levels and regulates inflammation by cox-2 and cell cycle arrest by p21 during lung carcinogenesis in mice. PMID- 21633291 TI - Research in emergency medicine in Europe. AB - Health research is fundamental for clinical excellence, a fact that applies equally to emergency medicine (EM). Although European scientific publication rates in EM have traditionally been lower than those of other medical specialties, from 1995 steady progress has been made. To increase the scientific output in EM it is necessary to resolve issues that hinder this progress, including the fact that EM is a new specialty, or even nonexistent in many European countries. This has resulted in a relative lack of scientific culture and training in research methodology of emergency physicians, of explicit recognition of scientific work, or of emergency physician competitiveness to apply to national and European grants for research projects. In addition, it is necessary to improve representation of European journals indexed in the category of EM and to receive a firm boost to EM research from the European Society for Emergency Medicine as well as from all European national societies. This study reviews these aspects and offers a personal perspective on where European EM research should be going. PMID- 21633292 TI - College of American Pathologists/American College of Medical Genetics proficiency testing for constitutional cytogenomic microarray analysis. AB - PURPOSE: To evaluate the feasibility of administering a newly established proficiency test offered through the College of American Pathologists and the American College of Medical Genetics for genomic copy number assessment by microarray analysis, and to determine the reproducibility and concordance among laboratory results from this test. METHODS: Surveys were designed through the Cytogenetic Resource Committee of the two colleges to assess the ability of testing laboratories to process DNA samples provided and interpret results. Supplemental questions were asked with each Survey to determine laboratory practice trends. RESULTS: Twelve DNA specimens, representing 2 pilot and 10 Survey challenges, were distributed to as many as 74 different laboratories, yielding 493 individual responses. The mean consensus for matching result interpretations was 95.7%. Responses to supplemental questions indicate that the number of laboratories offering this testing is increasing, methods for analysis and evaluation are becoming standardized, and array platforms used are increasing in probe density. CONCLUSION: The College of American Pathologists/American College of Medical Genetics proficiency testing program for copy number assessment by cytogenomic microarray is a successful and efficient mechanism for assessing interlaboratory reproducibility. This will provide laboratories the opportunity to evaluate their performance and assure overall accuracy of patient results. The high level of concordance in laboratory responses across all testing platforms by multiple facilities highlights the robustness of this technology. PMID- 21633293 TI - Quality improvement of mitochondrial respiratory chain complex enzyme assays using Caenorhabditis elegans. AB - PURPOSE: The diagnosis of a mitochondrial disorder relies heavily on the enzymatic analysis of mitochondrial respiratory chain complexes in muscle or other tissues. However, considerable differences exist between clinical laboratories in the protocols or particular tests used for evaluation. In addition, laboratories can encounter difficulties in consistent technique, as well as procurement of adequate positive or negative controls. Currently, there is no external quality assurance for respiratory chain complex assays. In this study, we explored the use of Caenorhabditis elegans mitochondria as a potential aid to diagnostic centers that perform respiratory chain complex assays. METHOD: Five diagnostic test centers in the United States and one from Australia comparatively analyzed enzyme activities of mitochondria from C. elegans. The first survey consisted of three open-labeled samples including one normal control and two mutants; the second survey consisted of one open-labeled normal control and two blinded samples. RESULTS: There was very good concordance among laboratories in detecting the majority of the defects present in the mutant specimens. Despite the ability to detect respiratory chain complex defects, the scatter between centers for certain enzymatic assays, particularly I + III, II, III, and IV, led to different diagnostic interpretations between the centers. CONCLUSION: The data strongly support the need for comparative testing of mitochondrial enzyme assays between multiple laboratories. Our overall results are encouraging for the use of nematode mitochondria as a tool that might provide a virtually inexhaustible supply of mitochondria with defined defects for development of assays and as a potential source of control specimens. PMID- 21633295 TI - The utility of peritoneal biopsy and omentectomy in the upstaging of apparent early ovarian cancer. AB - HYPOTHESIS: The hypothesis of this study is that routine blind peritoneal biopsies performed during the surgical staging of apparent early ovarian cancers rarely influence final cancer stage and thus are of little benefit to staging. Few studies have been done examining this question of whether the biopsies of grossly normal-appearing peritoneal tissue are of benefit to the surgical staging procedure. METHODS: Operative and pathology reports from 122 patients with early stage epithelial ovarian cancer staged by gynecologic oncologists at Barnes Jewish Hospital from 1995 to 2009 were reviewed. All had full surgical staging resulting in a final stage of IA to IIIA. The operative findings were assessed to determine how frequently the peritoneal biopsies upstaged the cancer. Other findings including age, grade, histological type, and preoperative CA-125 were assessed. RESULTS: The median age of the patients was 53 years (range, 23-81 years). The distribution of cancer types was endometrioid (42), serous (23), clear cell (19), mucinous (16), and mixed or other (22). The most frequent stage was IC (n = 50; 41%), followed by IA (n = 40; 33%). A total of 19 patients had positive peritoneal biopsies (16%). Of these, only 6 (5%) were microscopically positive, or from normal-appearing tissue. Five (4%) of these 6 subjects were upstaged by the random peritoneal biopsies alone. Five (4%) of the patients had microscopic metastases to the omentum, 4 (3%) of whom were upstaged by this finding alone. One patient had both microscopic peritoneal and omental disease. CONCLUSIONS: Although the rate of microscopic metastases to peritoneal tissue is low, random peritoneal biopsies are still indicated in early-stage disease owing to the low morbidity of the procedure and a small but present possibility of upstaging and altered management. Furthermore, systematic peritoneal biopsies ensure careful palpation and examination of all surfaces. PMID- 21633294 TI - Public attitudes toward ancillary information revealed by pharmacogenetic testing under limited information conditions. AB - PURPOSE: Pharmacogenetic testing can inform drug dosing and selection by aiding in estimating a patient's genetic risk of adverse response and/or failure to respond. Some pharmacogenetic tests may generate ancillary clinical information unrelated to the drug treatment question for which testing is done-an informational "side effect." We aimed to assess public interest and concerns about pharmacogenetic tests and ancillary information. METHODS: We conducted a random-digit-dial phone survey of a sample of the US public. RESULTS: We achieved an overall response rate of 42% (n = 1139). When the potential for ancillary information was presented, 85% (+/-2.82%) of respondents expressed interest in pharmacogenetic testing, compared with 82% (+/-3.02%) before discussion of ancillary information. Most respondents (89% +/- 2.27%) indicated that physicians should inform patients that a pharmacogenetic test may reveal ancillary risk information before testing is ordered. Respondents' interest in actually learning of the ancillary risk finding significantly differed based on disease severity, availability of an intervention, and test validity, even after adjusting for age, gender, education, and race. CONCLUSION: Under the limited information conditions presented in the survey, the potential of ancillary information does not negatively impact public interest in pharmacogenetic testing. Interest in learning ancillary information is well aligned with the public's desire to be informed about potential benefits and risks before testing, promoting patient autonomy. PMID- 21633296 TI - Reporting and validation of gynaecological Groupe Euopeen de Curietherapie European Society for Therapeutic Radiology and Oncology (ESTRO) brachytherapy recommendations for MR image-based dose volume parameters and clinical outcome with high dose-rate brachytherapy in cervical cancers: a single-institution initial experience. AB - OBJECTIVE: The objectives are to report the dosimetric analysis, preliminary clinical outcome, and comparison with published data of 3-dimensional magnetic resonance-based high dose rate brachytherapy (BT) in cervical cancer. MATERIALS AND METHODS: The data set of 24 patients with cervical cancer treated with high dose-rate brachytherapy applications was analyzed. All patients received radiation with or without chemotherapy (10 patients received concomitant chemoradiation). Point A, International Commission on Radiation Units and Measurement (ICRU) point doses, and Groupe Europeen de Curietherapie-European Society for Therapeutic Radiology and Oncology dose volume parameters, namely, high-risk clinical target volume (HR-CTV), D90 and D100 doses, and dose to D0.1cc and D2cc, for rectum, bladder, and sigmoid, were calculated and correlated. RESULTS: Mean +/- SD HR-CTV was 45.2 +/- 15.8 cc. The mean +/- SD point A dose was 73.4 +/- 4.5 Gy (median, 74.3 Gy) total biologically equivalent dose in 2 Gy per fraction (EQD2), whereas mean +/- SD D90 doses were 70.9 +/- 10.6 GyEQD2 (median, 68). The mean +/- SD ICRU rectal and bladder points were 63.5 +/- 8.1 and 80.4 +/- 34.4 GyEQD2, respectively. The D0.1cc and D2cc for rectum were 66.0 +/- 9.9 GyEQD2 (median, 64.5) and 57.8 +/- 7.7 GyEQD2 (median, 58.8), for bladder 139.1 +/- 54.7 GyEQD2 (median, 131.9) and 93.4 +/- 24.6 GyEQD2 (median, 91), and sigmoid were 109.4 +/- 45.2 GyEQD2 (median, 91) and 74.6 +/- 19.6 GyEQD2 (median, 69.6). With a median follow-up of 24 months, 3 patients had local nodal failure, 1 had right external iliac nodal failure, and 1 had left supraclavicular nodal failure. CONCLUSIONS: The 3-D magnetic resonance image-based high dose-rate brachytherapy approach in cervical cancers is feasible. In our experience, the HR CTV volumes are large, and D0.1cc and D2cc doses to bladder and sigmoid are higher than published literature so far. PMID- 21633297 TI - Human epididymis protein 4 (HE4) as a serum tumor biomarker in patients with ovarian carcinoma. AB - BACKGROUND: Ovarian cancer remains a leading cause of death from gynecological malignancy. Early diagnosis is the most important determinant of survival. For more than 25 years, cancer antigen 125 (CA 125) has been the criterion standard biomarker for the diagnosis and management of women with epithelial ovarian cancer. This study evaluated human epididymis protein 4 (HE4), a novel ovarian cancer biomarker, both alone and in combination with CA 125 as a diagnostic marker for ovarian cancer in a Chinese population. METHODS: Sera from 491 Chinese women with ovarian cancer or nonmalignant disorders and healthy women were analyzed. Sensitivities and specificities for both biomarkers and the combination were determined using predefined cutoffs (HE4>150 pmol/L and CA 125>35 U/mL) and receiver operator characteristic curves to define cutoffs based on 95% and 98% sensitivities. RESULTS: At baseline, serum HE4 and CA 125 levels were significantly higher in the ovarian cancer group versus the 5 reference groups. Using predefined cutoffs, HE4 specificity for ovarian cancer ranged from 90% to 100%; CA 125 specificity ranged from 36% (benign gynecologic disease) to 99%. Combining both markers yielded specificity for ovarian cancer of 100%. Using receiver operator characteristic curve analysis, the cutoff for 95% and 98% specificity was 102.6 and 150.2 pmol/L for HE4, respectively, and 127.2 and 325.5 U/mL for CA 125, respectively; the sensitivity of CA 125 for distinguishing ovarian cancer from benign gynecologic disease was 54% (95% specificity) and 28% (98% specificity), improving to 78% and 68%, respectively, with the addition of HE4. CONCLUSIONS: Human epididymis protein 4 used in conjunction with CA 125 yields improved specificity for ovarian cancer compared with the use of CA 125 alone, generally similar to results seen in non-Chinese populations. PMID- 21633298 TI - Expression of survivin, CD117, and C-erbB-2 in neuroendocrine carcinoma of the uterine cervix. AB - OBJECTIVE: To evaluate the expression and prognostic significance of survivin, CD117, and C-erbB-2 in neuroendocrine carcinoma of the uterine cervix. MATERIALS AND METHODS: Immunohistochemical stains of survivin, CD117, and C-erbB-2 were evaluated in 100 cases of cervical neuroendocrine carcinoma. The findings were correlated with clinicopathologic variables and disease-free survival. RESULTS: Expressions of survivin, CD117, and C-erbB-2 were detected in 27.0%, 12.0%, and 2.0% of the cases, respectively. Survivin-positive patients had a significantly younger mean age than the survivin-negative group (P=0.033). In early-stage cases, tumor recurrence was significantly associated with lymph node metastasis (P=0.005), depth of invasion (P=0.028), and the presence of lymphovascular space invasion (P=0.031) but not with the expression of survivin or CD117. Subgroup analysis in early-stage cases without lymph node metastasis (n=32) showed that only survivin expression had a significant association with decreased disease free survival (P=0.041). CONCLUSIONS: Survivin expression may be a prognostic indicator for survival in early-stage neuroendocrine carcinoma of the uterine cervix without lymph node metastasis. Adjuvant survivin-targeted therapy may have potential benefit in patients with this tumor. PMID- 21633299 TI - Prognostic significance of clusterin expression in advanced-stage cervical cancer treated with curative intended radiotherapy. AB - OBJECTIVE: Overexpression of clusterin (CLU), an antiapoptotic molecule, has been reported to induce resistance to radiotherapy (RT) in a variety of cancer cell types. The aim of this study was to evaluate the significance of CLU expression to predict survival of patients with advanced-stage cervical cancer who received curative intended RT. METHODS: Biopsy tissue specimens of advanced-stage cervical cancer before curative intended RT were obtained from 34 patients who were treated at Hokkaido University Hospital between 1998 and 2008 and whose complete medical records were available. The expression of CLU protein was analyzed by immunohistochemistry. Findings were evaluated in relation to several clinicopathological factors. Survival analyses were performed using the Kaplan Meier curves and the log-rank test. Independent prognostic factors were determined by multivariate Cox regression analysis. RESULTS: Clusterin protein was mainly present in the cytoplasm of cervical cancer cells. The expression of CLU protein in cervical cancer tissues before curative intended RT was not significantly related to any clinicopathological factors analyzed, including age, clinical stage, histologic type, and response to RT. Univariate analysis on prognostic factors showed that histologic type (P = 0.001), and CLU expression (P = 0.02) were related to survival. Multivariate analysis revealed that both histologic type (P = 0.002), and CLU expression (P = 0.02) were independent prognostic factors for overall survival. CONCLUSION: We conclude that CLU could be a new molecular marker to predict overall survival of patients with advanced stage cervical cancer treated with curative intended RT. PMID- 21633300 TI - Surgical and medical treatment of clear cell ovarian cancer: results from the multicenter Italian Trials in Ovarian Cancer (MITO) 9 retrospective study. AB - OBJECTIVE: Clear cell ovarian carcinoma has a poorer prognosis compared with other histological subtypes. MATERIALS AND METHODS: The Multicenter Italian Trials in Ovarian Cancer (MITO) 9 study retrospectively assessed an Italian cohort of patients with clear cell ovarian cancer observed in the years 1991-2007 in 20 Italian centers. RESULTS: A total of 240 patients with ovarian cancer were analyzed. Forty-five percent of the patients had stage I disease. In 62.9%, clear cell histology was pure, whereas in the other cases, a mixed population was evident. Most of the cases underwent standard surgery, whereas in 7.1% of the patients, a fertility-sparing surgery was given. Lymphadenectomy was performed in 47.9% (115/240) of the patients (54.3% in stages I and II; 39.2% in advanced stage). Most of the patients were treated with platinum-based chemotherapy including paclitaxel in 52.9%. Disease-free survival was longer in patients undergoing lymphadenectomy at surgery (P = 0.0001), both in early stages (P = 0.0258) and in stage III and IV diseases (P = 0.0037). The impact of lymphadenectomy was also evident on overall survival in patients with advanced stage disease. At multivariate analysis, lymphadenectomy (done vs not done) and stage (I and II vs III and IV) were independently associated with longer disease free and overall survival, whereas front-line chemotherapy (with vs without taxanes) was not significant. CONCLUSION: This analysis suggests that lymphadenectomy has a strong prognostic role for clear cell ovarian cancer influencing disease-free survival and overall survival. The addition of paclitaxel to platinum-based chemotherapy does not affect the outcome. PMID- 21633301 TI - Preoperative multimodal strategies for risk assessment of adnexal masses: analysis of 1362 cases in a gynecologic cancer center. AB - OBJECTIVE: Assessment of adnexal masses focuses on the accurate discrimination between benign and malignant lesions. In our tertiary referral center, we evaluated the discriminative power of expert sonography, serum CA-125 measurement, risk malignancy index (RMI) by Jacobs, and 2 preoperative triage strategies (combination of CA-125 measurement and RMI assessment with expert sonography). METHODS: From 2002 to 2008, a total of 1362 surgical explorations with indication of an adnexal mass from our department were included in this study. Preoperative workup in all patients comprised a gynecologic examination, expert sonography, and serum CA-125 measurement. We calculated sensitivity, specificity, positive and negative predictive value (PPV and NPV), and Cohen kappa (prevalence-adjusted measurement) to evaluate the discriminative power of each diagnostic test. RESULTS: Discriminative power of the evaluated tests differed depending on patients' menopausal state. In the premenopause, expert sonography reached the highest discriminative power with a kappa value of 0.53, a PPV of 0.45, and an NPV of 0.99. In the postmemopause, the combinations of expert sonography with CA-125 serum measurement or RMI assessment achieved the highest discriminative power: The combination of CA-125 and expert sonography reached a PPV of 0.89 and an NPV of 0.97; kappa yielded 0.84. The RMI combined with expert sonography as a triage strategy showed comparable results with a PPV of 0.89, an NPV of 0.96, and a kappa value of 0.82. CONCLUSIONS: Preoperative assessment of an adnexal mass may be guided by the patient's menopausal state. In premenopausal patients, expert sonography is helpful for preoperative differentiation between benign and malignant lesions; in postmenopausal patients, the use of triage strategies of either CA-125 serum measurement or RMI combined with expert sonography can be recommended. PMID- 21633302 TI - Expression of alphaV-integrins in uterine serous papillary carcinomas; implications for targeted therapy with intetumumab (CNTO 95), a fully human antagonist anti-alphaV-integrin antibody. AB - OBJECTIVE: Uterine serous papillary carcinoma (USPC) is an aggressive variant of endometrial cancer characterized by an innate resistance to chemotherapy and poor prognosis. In this study, we evaluated the expression of alphaV-integrins in primary USPC cell lines and the in vitro ability of intetumumab (CNTO 95), a fully human monoclonal antibody against alphaV-integrins, to inhibit USPC cell adhesion and migration. MATERIALS AND METHODS: The surface expression of integrins belonging to the alphaV-family, including alphaVbeta3, alphaVbeta5, and alphaVbeta6, was evaluated in 6 primary USPC cell lines using flow cytometry analysis. To test the ability of intetumumab to inhibit USPC cell adhesion and migration, adhesion assays in the presence of vitronectin and migration assays through an 8.0-MUm pore polycarbonate membrane also were performed. RESULTS: We found high expression of the alphaV-subunit on the cell surface of all 6 primary USPC cell lines tested (100% positive cells; mean fluorescence intensity range, 13.1-39.5). When the expression of single heterodimeric integrins was evaluated, alphaVbeta3, alphaVbeta5, and alphaVbeta6 were expressed on 37.5%, 32.0%, and 16.3% of cells (mean fluorescence intensity range, 6.5-16.2, 9.2-32.5, and 6.2 11.5, respectively). Importantly, in functional assays, low doses of intetumumab were effective in inhibiting adhesion (0.15 MUg/mL, P = 0.003) and migration (1.25 MUg/mL P = 0.02) of primary USPC cell lines. CONCLUSIONS: The alphaV integrins are overexpressed on the cell surface of primary USPC cell lines. Intetumumab may significantly inhibit USPC cell adhesion and migration pathways and may therefore represent a novel treatment option for patients harboring this rare but highly aggressive variant of endometrial cancer. PMID- 21633303 TI - Effects of bufalin on the proliferation of human choriocarcinoma cells. AB - OBJECTIVE: Bufalin is a traditional Chinese medicine, and it induces apoptosis in some lines of human tumor cells. METHODS: We investigated the effect of bufalin in the choriocarcinoma cell line, BeWo. BeWo cells were treated with various concentrations of bufalin, and changes in cell growth, the cell cycle, apoptosis, and related parameters were examined. RESULTS: An 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide assay showed that BeWo cells were sensitive to the growth inhibitory effect of bufalin. Cell cycle analysis indicated that exposure to bufalin decreased the proportion of cells in the synthesis phase and increased the proportion in the G0/G1 phases of the cell cycle. Induction of apoptosis was confirmed by annexin V staining of externalized phosphatidylserine and by the loss of mitochondrial transmembrane potential. This induction occurred in conjunction with the altered expression of genes related to cell growth, malignant phenotype, and apoptosis. CONCLUSIONS: These results suggest that bufalin may serve as a therapeutic agent for the treatment of choriocarcinoma. PMID- 21633304 TI - Defining prognostic variables in recurrent endometrioid endometrial cancer: a 15 year single-institution review. AB - OBJECTIVE: This study aimed to examine the pattern of recurrence in patients with endometrioid endometrial cancer and to identify clinically important prognostic factors in the recurrent population. METHODS: With institutional review board approval, a retrospective review identified 1061 patients who underwent primary surgery and treatment of endometrioid endometrial cancer at our institution from 1994 to 2007. Of this cohort, 77 (7.2%) patients developed a recurrence. Clinical factors were recorded, and Spearman correlation coefficients and chi test were used to determine associations between groups. Kaplan-Meier survival estimates and Cox proportional-hazards model were used to determine how prognostic variables affected survival after recurrence (RS) and overall survival (OS). RESULTS: Of 77 patients, site of recurrence was not available in 5 patients. The distribution of recurrence in the remaining 72 patients was as follows: isolated vaginal 18% (13/72), nonvaginal pelvic 12% (9/72), distant 31% (22/72), abdominal 24% (17/72), and nodal 15% (11/72). There was an overrepresentation of advanced stage (P < 0.001) and high-grade (P < 0.003) at presentation in the recurrent group. Median OS was 3.4 years and median RS was 1.3 years. Low-grade tumors, early stage, and those with less than 50% myometrial invasion were associated with a significant OS and RS advantage. Age-adjusted isolated vaginal recurrence presented with a 1.2-year RS survival advantage (P < 0.03). An age-adjusted Cox proportional hazard ratio model incorporating significant prognostic variables demonstrated that the only independent variable associated with worse OS and RS was increased histologic grade with a hazard ratio of 2.31 (95% confidence interval, 1.25-3.97) for RS and 2.44 (95% confidence interval, 1.41-4.62) for OS. CONCLUSIONS: Those patients with high-grade histology at the time of initial diagnosis manifest a decreased OS and RS, suggesting that the intrinsic biology of the tumor has the greatest prognostic importance. PMID- 21633305 TI - Preoperative bowel preparation in gynecologic oncology: a review of practice patterns and an impetus to change. AB - OBJECTIVES: Preoperative mechanical bowel preparation (MBP) is commonly used in gynecologic oncology (GO). We wished to assess the practice patterns and beliefs within the Society of Gynecologic Oncologists of Canada (GOC), review the literature on MBP as applicable to GO surgeries, and construct recommendations specific to our subspecialty. METHODS: A 23-question, 10-minute Internet survey was sent to 110 GOC members regarding their use, rationale, and understanding of the literature pertaining to MBP for GO surgeries. The historical justifications for using MBP before pelvic and abdominal surgery were identified through literature review. RESULTS: Half of respondents (48%) routinely order MBP despite acknowledgment in 77% that there was no good evidence to support its use. Use encompassed all cancer sites (53% ovary, 32% endometrial, 27% cervical, and 8% vulvar) and approaches (43% laparotomy and 29% laparoscopy/robotics). The most common reasons cited for ordering MBP were to decrease risk of anastomotic leak and improve visualization. In the last 5 years, use of MBP has decreased in most (77%) GOC respondents. Of all respondents, 71% felt that formal recommendations specific to the field of GO would be helpful. None of the arguments for using MBP could be justified in the literature. In contrast, common and often serious sequelae from MBP are frequently described. Admitted use of bowel preparation in other surgical specialties was even higher (53%-99%) than within the GOC. CONCLUSIONS: There is no literature to support the routine use of MBP in GO. Published recommendations (herein) should support and guide change in practice. PMID- 21633306 TI - Rate of port-site metastasis is uncommon in patients undergoing robotic surgery for gynecological malignancies. AB - OBJECTIVE: To describe the rate of port-site metastasis in patients who underwent robotic surgery for suspected gynecological malignancy. METHODS: Using a prospective database, we identified all patients who underwent robotic surgery performed by the Gynecologic Oncology service at 1 institution between December 2006 and March 2010. Records of patients with confirmed malignancy were reviewed for clinicopathological data and information about port-site metastasis. RESULTS: One hundred eighty-one patients met the inclusion criteria. The median age was 55.4 years (range, 19-82 years), and the median body mass index was 29.6 kg/m2 (range, 17.9-70.7 kg/m2). Port-site metastases were detected in 2 patients (1.1%) at 3 weeks (patient 1) and 11 months (patient 2) after surgery. Patient 1 underwent surgery for an adnexal mass, and pathological examination revealed gallbladder adenocarcinoma metastatic to the ovary. She had a recurrence in the right lateral abdominal wall robotic trocar site with concurrent metastases in the gallbladder fossa and liver. Patient 2 was diagnosed with adenocarcinoma of unclear (cervical vs endometrial) origin. Imaging showed metastases in pelvic and para-aortic lymph nodes. She underwent laparoscopy and was found intraoperatively to have gross disease on the right ovary. The patient underwent right salpingo oophorectomy and chemoradiation. She had residual disease in the cervix and subsequently underwent robotic hysterectomy and left salpingo-oophorectomy. Pathological examination revealed endometrial cancer. She had a recurrence at the transumbilical trocar site concurrent with retroperitoneal lymphadenopathy and carcinomatosis. There were no cases of isolated port-site metastasis. CONCLUSIONS: The rate of port-site metastasis after robotic surgery in women with gynecological cancer is low and similar to the rate for laparoscopic procedures. PMID- 21633307 TI - Rectosigmoid xanthoma as a predictive marker for sporadic rectosigmoid cancer. PMID- 21633308 TI - Mentorship programs for gastroenterology program directors: is there an unmet need? AB - GOALS: The purpose of this study was to determine whether a need exists for mentorship programs among gastroenterology (GI) fellowship program directors (PDs), to investigate specific areas where mentoring would be helpful, and to assess the willingness to establish mentoring relationships. BACKGROUND: There is no research regarding mentorship for GI fellowship PDs or associate/assistant program directors (APDs). Although some mentoring resources currently exist, it is not clear whether they fulfill the needs of PDs and APDs. STUDY: Mentorship needs were assessed using an electronic survey sent to GI PDs and APDs who subscribe to the American Gastroenterological Association PDs' list server. RESULTS: Fifty-nine GI PDs (47.6% completion rate) and 18 APDs returned the survey. Seventy-five percent of PDs and 78% of APDs thought a formal mentorship program would be beneficial to those starting their role. Content areas were identified where a mentor would be most helpful included Accreditation Council for Graduate Medical Education competencies, accreditation requirements, curriculum development, and site visit preparation. CONCLUSIONS: Current GI PDs and APDs felt a mentoring program would be beneficial, despite the availability of several resources including web sites and meetings. Our results suggest that there remains an unmet need for mentorship resources among GI PDs and APDs. PMID- 21633309 TI - Congenital anomaly of low insertion of cystic duct: endoscopic retrograde cholangiopancreatography findings and clinical significance. AB - BACKGROUND/AIM: Low insertion of cystic duct (LICD) may be problematic during cholecystectomy. This study was performed retrospectively to assess the prevalence of LICD and identify the risk factors of stone recurrence between LICD and non-LICD (NLICD) after removal of stones. METHODS: Between January 1999 and November 2005, 3546 patients received endoscopic retrograde cholangiopancreatography examination for suspicion of biliary tract diseases. The age and sex-matched group with NLICD was enrolled to compare the clinical differences with LICD group. LICD was defined as "the orifice level of the cystic duct being below the low third of the extrahepatic duct." Recurrence was defined as "patients suffering from cholangitis or biliary stones 1 year later after the first intervention." RESULTS: Of the enrolled 3546 patients (male/female=1821/1725), 191 (5.4%) had LICD. Excluding cases of malignancy, nonbiliary stones, and incomplete data, 122 LICD patients were available. Periampullary diverticula and positive bacterial culture from bile were less common in the LICD group than the NLICD group (P=0.045; P<0.001, respectively). Lower recurrent rate of common bile duct (CBD) stones in the recurrent cases were found in the LICD group compared with the NLICD group (P=0.024; P=0.039, respectively). Univariate analysis revealed that LICD [odds ratio (OR)=0.284; P=0.032] and CBD stones (OR=4.496; P=0.006) were significantly correlated to stone recurrence. CONCLUSIONS: Our study clearly demonstrated the prevalence (5.4%) of LICD in cases with suspicion of biliary tract disease based on endoscopic retrograde cholangiopancreatography. Notably, the strongest predictors, NLICD and CBD stones, appeared to result in the higher stone recurrence. PMID- 21633310 TI - Risk factors for nocturnal reflux in a large GERD cohort. AB - BACKGROUND: Nocturnal gastroesophageal reflux disease (GERD) has been associated with severe complications including erosive esophagitis, peptic stricture, and even esophageal adenocarcinoma and is known to cause a decreased health-related quality of life. However, there is limited information on independent predictors of nocturnal GERD. AIM: The aim of this study was to examine the relationship between nocturnal GERD and patient demographics, symptoms, clinical and endoscopic findings. METHODS: Consecutive patients presenting to the gastrointestinal endoscopy unit for evaluation of GERD symptoms undergoing index endoscopy were asked to complete a validated GERD questionnaire. Demographics, clinical features, and endoscopic findings were recorded. Nocturnal GERD was defined as awakening at night by heartburn or acid regurgitation. Patient factors were compared using chi and Mann-Whitney U test. All factors that were statistically significant (P<0.05) were entered into a stepwise logistic regression to evaluate for independent predictors of nocturnal GERD. RESULTS: Of 908 GERD patients evaluated, 665 (73.2%) reported nocturnal symptoms. The majority of the patients were male (93%) and White (83%) with a mean age of 57.2 years (SD 12.7). On univariate analysis, younger age, higher body mass index, daily heartburn, heartburn duration of >5 years, severe heartburn, daily regurgitation, regurgitation duration >5 years (all P<0.05), and presence of hiatal hernia (P=0.02) were significantly associated with the presence of nocturnal GERD. On multivariate analysis, severe heartburn [3.3 (2.1 to 5.1), P<0.01] and daily heartburn [1.5(1.1 to 2.3), P=0.03], daily regurgitation [2.2 (1.1 to 4.2), P=0.025], and regurgitation duration of >5 years [1.7 (1.2 to 2.4), P<0.01], and presence of hiatal hernia [1.4 (1.1 to 2.0), P=0.03] were all independent risk factors for nocturnal GERD. CONCLUSIONS: Results of this large prospective cohort study of GERD patients suggest that nocturnal symptoms are reported by 3 quarters of patients. Patients with frequent and severe daytime symptoms are significantly more likely to experience nocturnal symptoms. There was no significant difference in the prevalence of Barrett esophagus between patients with and without nocturnal GERD. Finally, presence of a hiatus hernia on endoscopy is also an independent risk factor for nocturnal GERD. PMID- 21633311 TI - Perioperative management of children with encephalocele: an institutional experience. AB - BACKGROUND: Encephaloceles are neural tube defects that are characterized by protrusion of the brain and meninges through a defect in the cranium. The inherent implications of pediatric anesthesia and difficult airway make surgical correction challenging for anesthesiologists METHODS: Available medical records of 118 children who underwent excision and repair of encephalocele over a period of 10 years were analyzed retrospectively. Data on associated anomalies, anesthetic management, perioperative complications, and outcome at discharge were reviewed. RESULTS: The average age of presentation was 1 year and 6 months. The most common site of lesion was the occiput (67%). Encephaloceles were giant (size of sac larger than the head) in 15.3% of children. Hydrocephalus was the most common complication (45.8%) and was predominantly associated in children with occipital encephaloceles (P=0.00). Difficult mask ventilation and intubation were encountered in 5.9% and 19.5% of children, respectively. In children with occipital encephalocele, the trachea was intubated commonly by direct laryngoscopy in the lateral position (47.5%). The average blood loss was 69.6+/ 13.2 mL, and 56 children required transfusion, the average being 13.2+/-9.6 mL/kg. Intraoperative hemodynamic disturbances and respiratory complications were observed in 21.1% and 13.5% of children, respectively. The mean intensive care unit and hospital stay were 1.8+/-2.1 and 8.6+/-4.9 days, respectively. The stays were prolonged significantly whenever the children developed hydrocephalus, meningitis, and respiratory infection, predisposing to poor outcome. CONCLUSIONS: Difficult airway is not the only concern in children with encephalocele, but associated congenital malformations, hydrocephalus, large size of sac, and hemodynamic disturbances all require careful consideration. PMID- 21633312 TI - Limited atlantooccipital and cervical range of motion in patients with familial dysautonomia. AB - Familial dysautonomia (FD) is a rare neurological disease with autosomal recessive inheritance and is associated with severe kyphoscoliosis. Investigators have reported subjective observation of decreased cervical motion and high rates of proximal instrumentation failure in this population. A radiographic study of sagittal plane cervical spine motion was performed with 15 patients with FD. Measurements were compared with normal values. Patients with FD had decreased sagittal motion at all cervical levels (P<0.05). Intervertebral translation was also decreased significantly at C3-C6. FD is associated with decreased sagittal motion in the cervical spine. PMID- 21633313 TI - Evaluation of hepatocyte growth factor plasmid therapeutic effect by 99mTc hexakis-2-methyoxy-isobutylisonitrile blood flow scintigraphy in a rat model of hind limb ischemia. AB - OBJECTIVE: This study was conducted in an attempt to use blood flow scintigraphy with 99mTc-hexakis-2-methyoxy-isobutylisonitrile (99mTc-MIBI) for the evaluation of the angiogenic effect of hepatocyte growth factor (HGF) plasmid in a rat model of hind limb ischemia. MATERIALS AND METHODS: The femoral artery of the left hind limb of each rat was ligated to create a model of hind limb ischemia. Three weeks later, HGF plasmid (1.5 mg/1.1 ml/body) or saline (1.1 ml/body) was administered intramuscularly into three sites of the ischemic hind limb. Two and 4 weeks after the treatment, blood flow through the hind limb was measured by 99mTc-MIBI scintigraphy. In addition, the number of capillary endothelial cells obtained by immunostaining for CD31 was counted. RESULTS: After 99mTc-MIBI scintigraphy in the HGF plasmid-treated group, the blood flow ratio increased significantly from the pretreatment ratio 63.8 to 73.4%, 2 weeks after treatment (P<0.05) and to 97.8%, 4 weeks after treatment (P<0.05). The number of CD31-positive endothelial cells was significantly higher in the HGF plasmid-treated group than in the control group. CONCLUSIONS: The experimental study using a rat model of hind limb ischemia showed usefulness of 99mTc-MIBI scintigraphy to evaluate the angiogenic effect of HGF plasmid treatment. PMID- 21633314 TI - Somatostatin receptor scintigraphy and chromogranin A assay in staging and follow up of patients with well-differentiated neuroendocrine tumors. AB - INTRODUCTION: Chromogranin A (CgA) assay and somatostatin receptor scintigraphy (SRS) are implemented in the standard workup of neuroendocrine tumors (NETs). The aim of this study was to assess the value of SRS and CgA in staging and follow-up patients with well-differentiated NETs. MATERIALS AND METHODS: A total of 88 consecutive patients with histologically confirmed well-differentiated NETs were included. General data such as sex, age, site of primary tumor and metastases, medication, and follow-up results, including CgA values, were gathered. The number of lesions on SRS were scored and categorized from 0 to 3 and the uptake was scored from 0 to 4. RESULTS: CgA values differed significantly between patients with and without clinical symptoms (P=0.028), a positive and negative SRS (P=0.005), the different SRS scores (P=0.002), the number of lesions (P=0.001), and the presence or absence of liver metastasis (P=0.003). The sensitivity, specificity, positive predictive value, and negative predictive value were 78, 93, 98, and 47% for SRS and 62, 100, 100, and 35% for CgA; however, by combining the test, all results improved. All patients (n=11) referred for routine follow-up had stable CgA values, whereas in one patient only the SRS score increased. In the group of patients with a suspicion on tumor progression during follow-up (n=14), CgA values increased in nine patients. In this group, the SRS score increased in two patients. CONCLUSION: Despite the higher sensitivity of SRS than of CgA in staging and restaging well differentiated NETs, both tests are required at the initial stage. Disease extent, symptoms, and liver metastasis have an impact on both SRS results and CgA values. CgA has an important value in the assessment of tumor progression during follow-up, whereas the role of SRS in the routine follow-up of well differentiated NETs is limited. PMID- 21633315 TI - Radiation doses to cohabitants of patients undergoing radioiodine ablation for thyroid cancer: poor compliance with radiation protection guidelines but low radiation exposure. AB - BACKGROUND: The drive to reduce hospital stay after radioiodine remnant ablation in patients with thyroid cancer may increase the risk of radiation exposure to family members. The aim of this study was to evaluate the key determinants of dose exposure to familial members, with particular reference to the degree of adherence to current radiation safety guidelines. METHODS: All participants prospectively received our standard departmental oral and written safety instructions, with a mandatory 3-day restriction period. The postmicturition radiation levels of treated patients were measured (at 1-m distance) at the time of discharge using a portable radiometer. The radiation exposure of cohabitants was assessed with an optically stimulated luminescence-based personal dosimeter during the 3 days after hospital discharge. A questionnaire was used to assess the adherence of relatives/cohabitants to radiation safety guidelines. RESULTS: A total of 38 patients with thyroid cancer and 48 household members were included. At 48 h post therapy, the patient's median emission at 1-m distance was 13.4 MUSv/h. The mean cumulative cohabitant exposure was 102 MUSv (<50-1000). A positive correlation between cohabitant radiation exposure and the radiation level of the patient was observed (P=0.016). This correlation was absent when the recommended guidelines were followed (P=0.56). Only 17 household members (35.4%) strictly followed the recommended guidelines, but dose exposures exceeded 0.3 mSv in only four cases, in which a mean of between 5.8 and 9.5 h were spent in close proximity to the patient in the first 3 days, including sleeping with treated patients in half of the cases. CONCLUSION: Despite poor compliance with safety guidelines, a short-stay protocol respects current legislation, and is applicable to most patients treated with 3.7 GBq for radioiodine remnant ablation. PMID- 21633316 TI - Resection for secondary malignancy of the pancreas. AB - OBJECTIVES: This study tried to clarify the role of pancreatic resection in the treatment of secondary malignancy with metastasis or local invasion to the pancreas in terms of surgical risk and survival benefit. METHODS: Data of secondary malignancy of the pancreas from our 19 patients and cases reported in the English literature were pooled together for analysis. RESULTS: There were 329 cases of resected secondary malignancy of the pancreas, including 241 cases of metastasis and 88 cases of local invasion. The most common primary tumor metastatic to the pancreas and amenable to resection was renal cell carcinoma (RCC) (73.9%). More than half (52.3%) of the primary cancers with local invasion to the pancreas were colon cancer, and nearly half (40.9%) were stomach cancer. The median metastatic interval was 84 months (7 years) for overall primary tumors and 108 months (9 years) for RCC. The 5-year survival for secondary malignancy of the pancreas after resection was 61.1% for metastasis and 58.9% for local invasion, with 72.8% for RCC metastasis, 69.0% for colon cancer, and 43.8% for stomach cancer with local invasion to the pancreas. CONCLUSIONS: Pancreatic resection should not be precluded for secondary malignancy of the pancreas because long-term survival could be achieved with acceptable surgical risk in selected patients. PMID- 21633317 TI - Dynamic quantitative evaluation of contrast-enhanced endoscopic ultrasonography in the diagnosis of pancreatic diseases. AB - OBJECTIVES: This study aimed to investigate the usefulness of contrast-enhanced endoscopic ultrasonography (EUS) with time-intensity curve (TIC) in differentiating pancreatic diseases. METHODS: Patients who underwent contrast enhanced EUS between January 2007 and June 2009 were analyzed retrospectively, including 48 with pancreatic ductal cancer (PC), 14 with autoimmune pancreatitis (AIP), 13 with mass-forming pancreatitis (MFP), and 16 with pancreatic endocrine tumor (PET). After intravenous injection of contrast agent, contrast imaging pattern, TIC-based quantitative evaluation, and diagnostic ability of EUS in combination with TIC to diagnose benignancy or malignancy were assessed. RESULTS: Hypovascular and heterogeneous pattern (42/48) in PC, isovascular and homogenous (21/27) in AIP and MFP, and hypervascular and rapid stained (16/16) in PET were observed. The echo intensity reduction rate from the peak at 1 minute was the greatest in PC followed by MFP, AIP, and PET (P < 0.05). The diagnostic accuracies based on contrast imaging pattern (84.0%) and TIC (88.0%) were higher than those based on B-mode imaging (82.6%) and dynamic computed tomography (81.3%). In EUS in combination with TIC, sensitivity, specificity, and accuracy rose up to 95.8%, 92.6%, and 94.7%, respectively. CONCLUSIONS: Contrast-enhanced EUS with the dynamic quantitative analysis preparing TIC increased the diagnostic accuracy for pancreatic diseases. PMID- 21633319 TI - Varicella arthritis in childhood: a case report and review of the literature. AB - Nonbacterial arthritis is a rare but well-recognized complication of acute varicella in children. Reported cases usually described monoarthritis of the knee that occurs at the onset of the rash or shortly after. Herein, we describe a case of arthritis of the hip that occurred in a 4-year-old girl 6 days before the onset of rash. The presence of varicella-zoster virus DNA in synovial fluid was confirmed by real-time polymerase chain reaction. A review of the literature reported 26 previous cases of aseptic arthritis due to varicella infection among children. PMID- 21633318 TI - Increased expression of DNA repair genes in invasive human pancreatic cancer cells. AB - OBJECTIVE: Pancreatic cancer was the fourth leading cause of cancer death in the United States in 2010. Recurrence of disease after resection occurs because of neoplastic cell survival. To better understand these highly aggressive cells, gene expression microarrays were performed. METHODS: Using the established lines HPAC and PANC1 and a Matrigel assay, genome expression arrays were performed to analyze patterns between invasive and total cells. RESULTS: Significant increases in the expression of genes related to DNA repair were observed. A number of the same genes also demonstrated an increase in expression when comparing bulk cells to a putative tumor-initiating cell (TIC) population. The TIC population was isolated using the spheroid technique, and compared with bulk cells, spheroid cells functionally repair breaks in DNA faster after challenge with the drug gemcitabine. Finally, using Oncomine, we observed a significant increase in DNA copy number of BRCA1 and RAD51 in tissue isolated from metastatic pancreatic cancer compared with tissue isolated from the primary site. CONCLUSIONS: From these data, we conclude that the most invasive cells within a pancreatic tumor are able to thrive because of their increased genomic stability. These cells have also been linked to the TIC population in a tumor. PMID- 21633320 TI - Absence seizures associated with efavirenz initiation. AB - Efavirenz, used in treating pediatric human immunodeficiency virus infection, has central nervous system side effects. We report on a 5-year-old girl with perinatally acquired human immunodeficiency virus infection, presenting with new onset absence seizures after starting treatment with efavirenz. Plasma efavirenz values were above therapeutic range. The child was homozygous for the CYP2B6 516T/T genotype, which is associated with poor efavirenz clearance. Seizures abated after efavirenz discontinuation. PMID- 21633321 TI - Adenosine A2(A) receptor gene polymorphism (1976C>T) affects coronary flow reserve response during vasodilator stress testing in patients with non ischemic dilated cardiomyopathy. AB - OBJECTIVES: Patients with non ischemic-dilated cardiomyopathy (DCM) are characterized by an activation of the adenosinergic system and reduced coronary flow reserve (CFR) evaluated by transthoracic Doppler echocardiography during vasodilator adenosinergic stress (dipyridamole administration). The aim of this study was to assess whether genetic polymorphisms (263C>T and 1976C>T) of the A2(A) receptor gene affect CFR response in patients with DCM. METHODS: We enrolled a group of 80 patients with DCM (55 male; age, 62+/-10.3 years) and 162 healthy volunteers (55 male; age, 45.1+/-9.5 years). Doppler-derived CFR (high dose dipyridamole coronary diastolic peak flow velocity to resting coronary peak flow velocity ratio) of distal left anterior descending artery was determined in DCM. A2(A) receptor genotyping was determined in all patients by polymerase chain reaction-restriction fragment length polymorphism analysis. The expression of A2(A) protein and mRNA was also assessed in healthy controls. RESULTS: The genotype distribution of the 263C>T (P=0.5) and 1976C>T (P=0.8) polymorphisms was not significantly different between patients and controls. Patients with 1976TT genotype had significantly lower CFR value than 1976CC patients (2.3+/-0.7, 2.0+/ 0.5 and 1.9+/-0.4, P<0.05 for CC, CT and TT genotypes, respectively). Controls who were heterozygous (P=0.02) or homozygous (P=0.001) for the T1976 allele showed a significant increase in A2(A) receptor protein. CONCLUSION: These data demonstrate that A2(A) 1976C>T polymorphism is associated with a blunted coronary vasodilatory response in patients with DCM, and support a direct consequences of this single nucleotide polymorphism for protein expression. Additional studies are needed to better define the functional role of this genetic variant as well as to clarify the potential clinical impact of genetics during pharmacological stress cardiac imaging. PMID- 21633322 TI - Genetic risk factors for type 2 diabetes mellitus and response to sulfonylurea treatment. AB - OBJECTIVE: After the identification of type 2 diabetes mellitus (T2DM) risk alleles from genome-wide association studies, models have been developed to identify subjects at high risk to develop T2DM. We hypothesize that a panel of 20 repeatedly associated T2DM risk alleles influences response to sulfonylureas (SUs). METHODS: Two hundred and seven incident SU (tolbutamide, glibenclamide, glimepiride, gliclazide) users with T2DM were recruited from four primary care centers. A genetic risk score per patient was calculated based on the number of risk-alleles. With this score, patients were categorized into three predefined genetic risk groups. The effect of the genetic risk group on the achievement of stable SU dose, prescribed stable SU dose, and time to stable SU dose was analyzed. RESULTS: Carriers of more than 17 T2DM risk alleles had a 1.7-fold reduced likelihood to achieve stable SU dose (P=0.044). No significant effect of the number of T2DM risk alleles on prescribed dose was found. Carriers of more than 17 T2DM risk alleles showed a marginally significant increased time to stable dose (hazard ratio: 0.81; 95% confidence interval, 0.75-1.01, P=0.058). CONCLUSION: T2DM risk alleles are associated with response to SUs in primary care T2DM patients. This suggests that individualization of T2DM treatment according to genetic profile may be an opportunity to improve clinical outcome. PMID- 21633323 TI - Variation in the glucocorticoid receptor gene (NR3C1) may be associated with corticosteroid dependency and resistance in children with Crohn's disease. AB - OBJECTIVES: In pediatric onset of Crohn's disease (CD), corticosteroid dependency (approximately 40%) and resistance (approximately 10%) are significant clinical problems. Given the known effects of the glucocorticoid receptor (GR/NR3C1) gene in corticosteroid metabolism, we investigated whether variation in the gene was associated with corticosteroid response. METHODS: A retrospective cohort study was carried out including patients with CD diagnosed before 18 years and treated with a first course of corticosteroids in two Canadian tertiary pediatric gastroenterology clinics. DNA was obtained from blood or saliva. Tagging single nucleotide polymorphisms (SNPs) and functionally important SNPs were genotyped. Allelic, genotype, and haplotype associations between the glucocorticoid receptor SNPs and response to corticosteroids were examined. RESULTS: A total of 296 corticosteroid-resistant, corticosteroids-dependent, and corticosteroid responsive patients with CD were studied. Of the 12 SNPs examined, four markers, rs6196 [odds ratio (OR)=2.03; 95% confidence interval (CI): 1.03-4.0; P=0.042], rs7701443 (OR=3.43; 95% CI: 1.79-6.57; P=0.042), rs6190 (OR=4.84; 95% CI: 1.70 13.80; P=0.003), and rs860457 (OR=3.43; 95% CI: 1.79-6.57; P<0.001) were associated at the allelic level with corticosteroid resistance. Haplotype analysis of four associated markers revealed associations between two haplotypes and corticosteroid resistance (P values of 0.046 and 0.001). Three SNPs, rs10482682 (OR=1.43; 95% CI: 0.99-2.08; P=0.047), rs6196 (OR=0.55; 95% CI: 0.31 0.95; P=0.024), and rs2963155 (OR=0.64; 95% CI: 0.42-0.98; P=0.039), showed associations under an additive model, whereas rs4912911 (OR=0.37; 95% CI: 0.13 1.00; P=0.03) and rs2963156 (OR=0.32; 95% CI: 0.07-1.12; P=0.047) showed associations under a recessive model with corticosteroid dependence. Two five marker haplotypes were associated with corticosteroid dependence (P values 0.002 and 0.004). CONCLUSION: Our results suggest that variations in the GR/NR3C1 gene are associated with corticosteroid resistance and dependency in pediatric-onset CD. Studies are required to replicate these findings and to identify the potentially relevant variants. PMID- 21633324 TI - Gene transfer for ischemic heart failure in a preclinical model. AB - Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine their efficacy and safety. Gene therapy using viral vectors is one of the most promising approaches for treating cardiac diseases. Viral delivery of various different genes by changing the carrier gene has immeasurable therapeutic potential. In this video, the full process of an animal model of heart failure creation followed by gene transfer is presented using a swine model. First, myocardial infarction is created by occluding the proximal left anterior descending coronary artery. Heart remodeling results in chronic heart failure. Unique to our model is a fairly large scar which truly reflects patients with severe heart failure who require aggressive therapy for positive outcomes. After myocardial infarct creation and development of scar tissue, an intracoronary injection of virus is demonstrated with simultaneous nitroglycerine infusion. Our injection method provides simple and efficient gene transfer with enhanced gene expression. This combination of a myocardial infarct swine model with intracoronary virus delivery has proven to be a consistent and reproducible methodology, which helps not only to test the effect of individual gene, but also compare the efficacy of many genes as therapeutic candidates. PMID- 21633325 TI - A protocol for collecting and staining hemocytes from the yellow fever mosquito Aedes aegypti. AB - Mosquitoes are vectors for a number of disease-causing pathogens such as the yellow fever virus, malaria parasites and filarial worms. Laboratories are investigating anti-pathogen components of the innate immune system in disease vector species in the hopes of generating transgenic mosquitoes that are refractory to such pathogens(1, 2). The innate immune system of mosquitoes consists of several lines of defense (3). Pathogens that manage to escape the barrier imposed by the epithelium-lined mosquito midgut (4) enter the hemolymph and encounter circulating hemocytes, important cellular components that encapsulate and engulf pathogens (5, 6). Researchers have not found evidence for hematopoietic tissues in mosquitoes and current evidence suggests that the number of hemocytes is fixed at adult emergence and numbers may actually decline as the mosquito ages (7). The ability to properly collect and identify hemocytes from medically important insects is an essential step for studies in cellular immunity. However, the small size of mosquitoes and the limited volume of hemolymph pose a challenge to collecting immune cells. Two established methods for collecting mosquito hemocytes include expulsion of hemolymph from a cut proboscis (8), and volume displacement (perfusion), in which saline is injected into the membranous necklike region between the head and thorax (i.e., cervix) and the perfused hemolymph is collected from a torn opening in a distal region of the abdomen (9, 10). These techniques, however, are limited by low recovery of hemocytes and possible contamination by fat body cells, respectively (11). More recently a method referred to as high injection/recovery improved recovery of immunocytes by use of anticoagulant buffers while reducing levels of contaminating scales and internal tissues (11). While that method allows for an improved method of collecting and maintaining hemocytes for primary culture, it entails a number of injection and collecting steps that are not necessary if the downstream goal is to collect, fix and stain hemocytes for diagnostics. Here, we demonstrate our method of collecting mosquito hemolymph that combines the simplicity of perfusion, using anticoagulant buffers in place of saline solution, with the accuracy of high injection techniques to isolate clean preparations of hemocytes in Aedes mosquitoes. PMID- 21633326 TI - In vitro assay of bacterial adhesion onto mammalian epithelial cells. AB - To cause infections, bacteria must colonize their host. Bacterial pathogens express various molecules or structures able to promote attachment to host cells(1). These adhesins rely on interactions with host cell surface receptors or soluble proteins acting as a bridge between bacteria and host. Adhesion is a critical first step prior to invasion and/or secretion of toxins, thus it is a key event to be studied in bacterial pathogenesis. Furthermore, adhered bacteria often induce exquisitely fine-tuned cellular responses, the studies of which have given birth to the field of 'cellular microbiology'(2). Robust assays for bacterial adhesion on host cells and their invasion therefore play key roles in bacterial pathogenesis studies and have long been used in many pioneer laboratories(3,4). These assays are now practiced by most laboratories working on bacterial pathogenesis. Here, we describe a standard adherence assay illustrating the contribution of a specific adhesin. We use the Escherichia coli strain 2787(5), a human pathogenic strain expressing the autotransporter Adhesin Involved in Diffuse Adherence (AIDA). As a control, we use a mutant strain lacking the aidA gene, 2787DeltaaidA (F. Berthiaume and M. Mourez, unpublished), and a commercial laboratory strain of E. coli, C600 (New England Biolabs). The bacteria are left to adhere to the cells from the commonly used HEp-2 human epithelial cell line. This assay has been less extensively described before(6). PMID- 21633327 TI - Organotypic cerebellar cultures: apoptotic challenges and detection. AB - Organotypic cultures of neuronal tissue were first introduced by Hogue in 1947 (1,2) and have constituted a major breakthrough in the field of neuroscience. Since then, the technique was developed further and currently there are many different ways to prepare organotypic cultures. The method presented here was adapted from the one described by Stoppini et al. for the preparation of the slices and from Gogolla et al. for the staining procedure (3,4). A unique feature of this technique is that it allows you to study different parts of the brain such as hippocampus or cerebellum in their original structure, providing a big advantage over dissociated cultures in which all the cellular organization and neuronal networks are disrupted. In the case of the cerebellum it is even more advantageous because it allows the study of Purkinje cells, extremely difficult to obtain as dissociated primary culture. This method can be used to study certain developmental features of the cerebellum in vitro, as well as for electrophysiological and pharmacological experiments in both wild type and mutant mice. The method described here was designed to study the effect of apoptotic stimuli such as Fas ligand in the developing cerebellum, using TUNEL staining to measure apoptotic cell death. If TUNEL staining is combined with cell type specific markers, such as Calbindin for Purkinje cells, it is possible to evaluate cell death in a cell population specific manner. The Calbindin staining also serves the purpose of evaluating the quality of the cerebellar cultures. PMID- 21633328 TI - Hyponeophagia: a measure of anxiety in the mouse. AB - Before the present day, when fast-acting and potent rodenticides such as alpha chloralose were not yet in use, the work of pest controllers was often hampered by a phenomenon known as "bait shyness". Mice and rats cannot vomit, due to the tightness of the cardiac sphincter of the stomach, so to overcome the problem of potential food toxicity they have evolved a strategy of first ingesting only very small amounts of novel substances. The amounts ingested then gradually increase until the animal has determined whether the substance is safe and nutritious. So the old rat-catchers would first put a palatable substance such as oatmeal, which was to be the vehicle for the toxin, in the infested area. Only when large amounts were being readily consumed would they then add the poison, in amounts calculated not to affect the taste of the vehicle. The poisoned bait, which the animals were now readily eating in large amounts, would then swiftly perform its function. Bait shyness is now used in the behavioural laboratory as a way of measuring anxiety. A highly palatable but novel substance, such as sweet corn, nuts or sweetened condensed milk, is offered to the mice (or rats) in a novel situation, such as a new cage. The latency to consume a defined amount of the new food is then measured. PMID- 21633329 TI - Rejection of fluorescence background in resonance and spontaneous Raman microspectroscopy. AB - Raman spectroscopy is often plagued by a strong fluorescent background, particularly for biological samples. If a sample is excited with a train of ultrafast pulses, a system that can temporally separate spectrally overlapping signals on a picosecond timescale can isolate promptly arriving Raman scattered light from late-arriving fluorescence light. Here we discuss the construction and operation of a complex nonlinear optical system that uses all-optical switching in the form of a low-power optical Kerr gate to isolate Raman and fluorescence signals. A single 808 nm laser with 2.4 W of average power and 80 MHz repetition rate is split, with approximately 200 mW of 808 nm light being converted to < 5 mW of 404 nm light sent to the sample to excite Raman scattering. The remaining unconverted 808 nm light is then sent to a nonlinear medium where it acts as the pump for the all-optical shutter. The shutter opens and closes in 800 fs with a peak efficiency of approximately 5%. Using this system we are able to successfully separate Raman and fluorescence signals at an 80 MHz repetition rate using pulse energies and average powers that remain biologically safe. Because the system has no spare capacity in terms of optical power, we detail several design and alignment considerations that aid in maximizing the throughput of the system. We also discuss our protocol for obtaining the spatial and temporal overlap of the signal and pump beams within the Kerr medium, as well as a detailed protocol for spectral acquisition. Finally, we report a few representative results of Raman spectra obtained in the presence of strong fluorescence using our time-gating system. PMID- 21633330 TI - Transplantation of cells directly into the kidney of adult zebrafish. AB - Regenerative medicine based on the transplantation of stem or progenitor cells into damaged tissues has the potential to treat a wide range of chronic diseases. However, most organs are not easily accessible, necessitating the need to develop surgical methods to gain access to these structures. In this video article, we describe a method for transplanting cells directly into the kidney of adult zebrafish, a popular model to study regeneration and disease. Recipient fish are pre-conditioned by irradiation to suppress the immune rejection of the injected cells. We demonstrate how the head kidney can be exposed by a lateral incision in the flank of the fish, followed by the injection of cells directly in to the organ. Using fluorescently labeled whole kidney marrow cells comprising a mixed population of renal and hematopoietic precursors, we show that nephron progenitors can engraft and differentiate into new renal tissue--the gold standard of any cell-based regenerative therapy. This technique can be adapted to deliver purified stem or progenitor cells and/or small molecules to the kidney as well as other internal organs and further enhances the zebrafish as a versatile model to study regenerative medicine. PMID- 21633331 TI - In vivo laser axotomy in C. elegans. AB - Neurons communicate with other cells via axons and dendrites, slender membrane extensions that contain pre- or post-synaptic specializations. If a neuron is damaged by injury or disease, it may regenerate. Cell-intrinsic and extrinsic factors influence the ability of a neuron to regenerate and restore function. Recently, the nematode C. elegans has emerged as an excellent model organism to identify genes and signaling pathways that influence the regeneration of neurons(1-6). The main way to initiate neuronal regeneration in C. elegans is laser-mediated cutting, or axotomy. During axotomy, a fluorescently-labeled neuronal process is severed using high-energy pulses. Initially, neuronal regeneration in C. elegans was examined using an amplified femtosecond laser(5). However, subsequent regeneration studies have shown that a conventional pulsed laser can be used to accurately sever neurons in vivo and elicit a similar regenerative response(1,3,7). We present a protocol for performing in vivo laser axotomy in the worm using a MicroPoint pulsed laser, a turnkey system that is readily available and that has been widely used for targeted cell ablation. We describe aligning the laser, mounting the worms, cutting specific neurons, and assessing subsequent regeneration. The system provides the ability to cut large numbers of neurons in multiple worms during one experiment. Thus, laser axotomy as described herein is an efficient system for initiating and analyzing the process of regeneration. PMID- 21633332 TI - High-throughput screening and biosensing with fluorescent C. elegans strains. AB - High-throughput screening (HTS) is a powerful approach for identifying chemical modulators of biological processes. However, many compounds identified in screens using cell culture models are often found to be toxic or pharmacologically inactive in vivo(1-2). Screening in whole animal models can help avoid these pitfalls and streamline the path to drug development. C. elegans is a multicellular model organism well suited for HTS. It is small (<1 mm) and can be economically cultured and dispensed in liquids. C. elegans is also one of the most experimentally tractable animal models permitting rapid and detailed identification of drug mode-of-action(3). We describe a protocol for culturing and dispensing fluorescent strains of C. elegans for high-throughput screening of chemical libraries or detection of environmental contaminants that alter the expression of a specific gene. Large numbers of developmentally synchronized worms are grown in liquid culture, harvested, washed, and suspended at a defined density. Worms are then added to black, flat-bottomed 384-well plates using a peristaltic liquid dispenser. Small molecules from a chemical library or test samples (e.g., water, food, or soil) can be added to wells with worms. In vivo, real-time fluorescence intensity is measured with a fluorescence microplate reader. This method can be adapted to any inducible gene in C. elegans for which a suitable reporter is available. Many inducible stress and developmental transcriptional pathways are well defined in C. elegans and GFP transgenic reporter strains already exist for many of them(4). When combined with the appropriate transgenic reporters, our method can be used to screen for pathway modulators or to develop robust biosensor assays for environmental contaminants. We demonstrate our C. elegans culture and dispensing protocol with an HTS assay we developed to monitor the C. elegans cap 'n' collar transcription factor SKN-1. SKN-1 and its mammalian homologue Nrf2 activate cytoprotective genes during oxidative and xenobiotic stress(5-10). Nrf2 protects mammals from numerous age related disorders such as cancer, neurodegeneration, and chronic inflammation and has become a major chemotherapeutic target(11-13).Our assay is based on a GFP transgenic reporter for the SKN-1 target gene gst-4(14), which encodes a glutathione-s transferase(6). The gst-4 reporter is also a biosensor for xenobiotic and oxidative chemicals that activate SKN-1 and can be used to detect low levels of contaminants such as acrylamide and methyl-mercury(15-16). PMID- 21633333 TI - Swimming performance assessment in fishes. AB - Swimming performance tests of fish have been integral to studies of muscle energetics, swimming mechanics, gas exchange, cardiac physiology, disease, pollution, hypoxia and temperature. This paper describes a flexible protocol to assess fish swimming performance using equipment in which water velocity can be controlled. The protocol involves one to several stepped increases in flow speed that are intended to cause fish to fatigue. Step speeds and their duration can be set to capture swimming abilities of different physiological and ecological relevance. Most frequently step size is set to determine critical swimming velocity (U(crit;)), which is intended to capture maximum sustained swimming ability. Traditionally this test has consisted of approximately ten steps each of 20 min duration. However, steps of shorter duration (e.g. 1 min) are increasingly being utilized to capture acceleration ability or burst swimming performance. Regardless of step size, swimming tests can be repeated over time to gauge individual variation and recovery ability. Endpoints related to swimming such as measures of metabolic rate, fin use, ventilation rate, and of behavior, such as the distance between schooling fish, are often included before, during and after swimming tests. Given the diversity of fish species, the number of unexplored research questions, and the importance of many species to global ecology and economic health, studies of fish swimming performance will remain popular and invaluable for the foreseeable future. PMID- 21633334 TI - Whole-mount immunohistochemical analysis for embryonic limb skin vasculature: a model system to study vascular branching morphogenesis in embryo. AB - Whole-mount immunohistochemical analysis for imaging the entire vasculature is pivotal for understanding the cellular mechanisms of branching morphogenesis. We have developed the limb skin vasculature model to study vascular development in which a pre-existing primitive capillary plexus is reorganized into a hierarchically branched vascular network. Whole-mount confocal microscopy with multiple labelling allows for robust imaging of intact blood vessels as well as their cellular components including endothelial cells, pericytes and smooth muscle cells, using specific fluorescent markers. Advances in this limb skin vasculature model with genetic studies have improved understanding molecular mechanisms of vascular development and patterning. The limb skin vasculature model has been used to study how peripheral nerves provide a spatial template for the differentiation and patterning of arteries. This video article describes a simple and robust protocol to stain intact blood vessels with vascular specific antibodies and fluorescent secondary antibodies, which is applicable for vascularized embryonic organs where we are able to follow the process of vascular development. PMID- 21633335 TI - Creation of reversible cholestatic rat model. AB - Cholestasis is a clinical condition commonly encountered by both surgeons and gastroenterologists. Cholestasis can cause various physiological changes and affect the nutritional status and surgical outcomes. Study of the pathophysiological changes occurring in the liver and other organs is of importance. Various studies have been done in cholestatic rat models. We used a reversible cholestatic rat model in our recent study looking at the role of methylprednisolone in the ischemia reperfusion injury. Various techniques for creation of a reversible cholestatic model have been described. Creation of a reversible cholestatic rat model can be challenging in view of the smaller size and unique hepatopancreatobiliary anatomy in rats. This video article demonstrates the creation of a reversible cholestatic model. This model can be used in various studies, such as looking at the changes in nutritional, physiological, pathological, histological and immunological changes in the gastrointestinal tract. This model can also be used to see the effects of cholestasis and various therapeutic interventions on major hepatic surgeries. PMID- 21633336 TI - Lentivirus-mediated genetic manipulation and visualization of olfactory sensory neurons in vivo. AB - Development of a precise olfactory circuit relies on accurate projection of olfactory sensory neuron (OSN) axons to their synaptic targets in the olfactory bulb (OB). The molecular mechanisms of OSN axon growth and targeting are not well understood. Manipulating gene expression and subsequent visualizing of single OSN axons and their terminal arbor morphology have thus far been challenging. To study gene function at the single cell level within a specified time frame, we developed a lentiviral based technique to manipulate gene expression in OSNs in vivo. Lentiviral particles are delivered to OSNs by microinjection into the olfactory epithelium (OE). Expression cassettes are then permanently integrated into the genome of transduced OSNs. Green fluorescent protein expression identifies infected OSNs and outlines their entire morphology, including the axon terminal arbor. Due to the short turnaround time between microinjection and reporter detection, gene function studies can be focused within a very narrow period of development. With this method, we have detected GFP expression within as few as three days and as long as three months following injection. We have achieved both over-expression and shRNA mediated knock-down by lentiviral microinjection. This method provides detailed morphologies of OSN cell bodies and axons at the single cell level in vivo, and thus allows characterization of candidate gene function during olfactory development. PMID- 21633337 TI - New paradigms to explain metastasis. PMID- 21633338 TI - PKMzeta maintains drug reward and aversion memory in the basolateral amygdala and extinction memory in the infralimbic cortex. AB - The intense associative memories that develop between drug-paired contextual cues and rewarding stimuli or the drug withdrawal-associated aversive feeling have been suggested to contribute to the high rate of relapse. Various studies have elucidated the mechanisms underlying the formation and expression of drug-related cue memories, but how this mechanism is maintained is unknown. Protein kinase M zeta (PKMzeta) was recently shown to be necessary and sufficient for long-term potentiation maintenance and memory storage. In the present study, we used conditioned place preference (CPP) and aversion (CPA) to examine whether PKMzeta maintains both morphine-associated reward memory and morphine withdrawal associated aversive memory in the basolateral amygdala (BLA). We also investigate the role of PKMzeta in the infralimbic cortex in the extinction memory of morphine reward-related cues and morphine withdrawal-related aversive cues. We found that intra-BLA but not central nucleus of the amygdala injection of the selective PKMzeta inhibitor ZIP 1 day after CPP and CPA training impaired the expression of CPP and CPA 1 day later, and the effect of ZIP on memory lasted at least 2 weeks. Inhibiting PKMzeta activity in the infralimbic cortex, but not prelimbic cortex, disrupted the expression of the extinction memory of CPP and CPA. These results indicate that PKMzeta in the BLA is required for the maintenance of associative morphine reward memory and morphine withdrawal associated aversion memory, and PKMzeta in the infralimbic cortex is required for the maintenance of extinction memory of morphine reward-related cues and morphine withdrawal-related aversive cues. PMID- 21633339 TI - Basolateral amygdala-driven augmentation of medial prefrontal cortex GABAergic neurotransmission in response to environmental stimuli associated with cocaine administration. AB - Basolateral amygdala (BLA) and medial prefrontal cortex (mPFC) interactions have been implicated in cue-elicited craving and drug seeking. However, the neurochemical mechanisms underlying drug/environment associations are ill defined. We used in vivo microdialysis and pharmacological inactivation techniques to identify alterations in mPFC glutamate (GLU) and gamma-aminobutyric acid (GABA) transmission in response to cues previously associated with experimenter-administered cocaine (COC) and the BLA contribution to these effects. Rats received alternate day injections of COC and saline (SAL) paired with a distinct environment for 6 days. Behavioral, neurochemical and immunohistochemical studies were conducted, in drug-free animals, 24 h after the last conditioning session. Animals exposed to a COC-paired environment demonstrated an augmented locomotor activity (LMA) relative to those exposed to the SAL-paired environment. mPFC GABA neurotransmission in the COC-paired environment was significantly increased, whereas GLU overflow was unaltered. Dual labeling of cFos and glutamic acid decarboxylase 67 immunoreactivity in mPFC neurons revealed significantly greater colocalization of these proteins following exposure to the COC-associated environment (CAE) relative to pseudo-conditioned rats or rats exposed to the SAL-associated environment indicating that the conditioned neurochemical response to the COC-paired environment is associated with activation of intrinsic mPFC GABA neurons. BLA inactivation prevented the increase in LMA and the augmentation of mPFC GABA transmission produced by cue exposure. Intra-mPFC application of the AMPA/KA receptor antagonist, NBQX, produced similar effects. These findings indicate that exposure to a CAE increases mPFC GABA transmission by enhancing excitatory drive from the BLA and activation of AMPA/KA receptors on mPFC GABA neurons. PMID- 21633340 TI - Pharmacokinetic impact of SLCO1A2 polymorphisms on imatinib disposition in patients with chronic myeloid leukemia. AB - The purpose of this study was to explore the role of the organic anion transporting polypeptide (OATP) 1A2, which is encoded by SLCO1A2, in the cellular uptake of the Bcr-Abl tyrosine kinase inhibitor imatinib, and the relationship between SLCO1A2 polymorphisms and the pharmacokinetics of imatinib in patients with chronic myeloid leukemia (CML). Imatinib uptake was significantly enhanced in OATP1A2-transfected human embryonic kidney (HEK) 293 cells (P = 0.002). Naringin, an OATP1A2 inhibitor, decreased the transport of imatinib in OATP1A2 transfected HEK293 cells, the human intestinal cell line Caco-2, and K562 CML cells. Linkage disequilibrium was found between the SLCO1A2 -1105G>A and -1032G>A genotypes in 34 CML patients and 100 healthy subjects. Imatinib clearance in CML patients was influenced by the SLCO1A2 -1105G>A/-1032G>A genotype (P = 0.075) and the SLCO1A2 -361GG genotype (P = 0.005). These findings suggest that imatinib is transported into cells by OATP1A2, and that SLCO1A2 polymorphisms significantly affect imatinib pharmacokinetics. PMID- 21633341 TI - Cigarette smoking, nicotine, and body weight. PMID- 21633342 TI - Dietary management of the metabolic syndrome. PMID- 21633343 TI - Mechanisms and genetics of antipsychotic-associated weight gain. PMID- 21633344 TI - The male genital tract is not a pharmacological sanctuary from efavirenz. AB - Many antiretroviral (ARV) drugs have large blood plasma-to-seminal plasma (BP/SP) concentration ratios. Concern exists that these drugs do not adequately penetrate the male genital tract (MGT), resulting in the MGT becoming a "pharmacological sanctuary" from these agents, with ineffective MGT concentrations despite effective blood concentrations. Efavirenz (EFV) is the most highly protein-bound ARV drug, with >99% binding in blood plasma and the largest BP/SP total EFV concentration ratio, reportedly ranging from 11 to 33. To evaluate protein binding as an explanation for the differences between the drug concentrations in blood and semen, we developed a novel ultrafiltration method, corrected for the duration of centrifugation, to measure protein binding in the two matrices. In six subjects, protein-free EFV concentrations were the same in blood and semen; the median (interquartile range (IQR)) protein-free EFV SP/BP ratio was 1.21 (0.99-1.35); EFV protein binding was 99.82% (99.79-99.86) in BP and 95.26% (93.24 96.67) in SP. This shows that the MGT is not a sanctuary from EFV. PMID- 21633345 TI - Implications of obesity for drug therapy: limitations and challenges. AB - Obesity has become a worldwide challenge with significant health and socioeconomic implications. One of the major implications is its impact on drug therapy. In order to gain a better understanding of this impact, we surveyed the regulatory guidances, the newly approved molecular entity drug products, and drug product labels in the Physician's Desk Reference. This review summarizes the findings of the survey along with the existing knowledge on pharmacokinetic and pharmacodynamic changes associated with obesity. PMID- 21633346 TI - Population pharmacokinetic-pharmacodynamic analysis of trastuzumab-associated cardiotoxicity. AB - Trastuzumab treatment is associated with cardiac dysfunction. The aim of this study was to develop a population pharmacokinetic-pharmacodynamic (PK-PD) model for the relationship between left ventricular ejection fraction (LVEF) and trastuzumab exposure and to identify associated clinically relevant covariates. Data from an unselected cohort of patients with early and advanced HER2-positive breast cancer receiving treatment with trastuzumab were analyzed using a nonlinear mixed-effects modeling approach. LVEF values from 240 patients were available. The data were best described by an effect-compartment model. The population LVEF recovery half-life after trastuzumab treatment (T(1/2rec)) was estimated at 49.7 days. The cumulative anthracycline dose was a significant determinant of the half maximal effect concentration (EC50), causing a 45.9% increase in sensitivity (decrease in EC50) at the maximum cumulative anthracycline dose. The developed population PK-PD model may be used to establish optimal treatment and cardiac monitoring strategies for trastuzumab. PMID- 21633347 TI - Should I say or should I stop: raising your voice to influence science policy. PMID- 21633348 TI - Wnt signalling escapes to cilia. AB - The primary cilium is proposed to restrain the level of canonical Wnt signalling, but it was unknown how the cilium achieves this. beta-catenin, a component of the canonical Wnt signalling pathway, is now shown to be sequestered to the cilium by the Wnt signalling modulator Jouberin (Jbn) to restrain Wnt responses. PMID- 21633354 TI - Condensin association with histone H2A shapes mitotic chromosomes. AB - Chromosome structure is dynamically regulated during cell division, and this regulation is dependent, in part, on condensin. The localization of condensin at chromosome arms is crucial for chromosome partitioning during anaphase. Condensin is also enriched at kinetochores but its precise role and loading machinery remain unclear. Here we show that fission yeast (Schizosaccharomyces pombe) kinetochore proteins Pcs1 and Mde4--homologues of budding yeast (Saccharomyces cerevisiae) monopolin subunits and known to prevent merotelic kinetochore orientation--act as a condensin 'recruiter' at kinetochores, and that condensin itself may act to clamp microtubule binding sites during metaphase. In addition to the regional recruitment factors, overall condensin association with chromatin is governed by the chromosomal passenger kinase Aurora B. Aurora-B-dependent phosphorylation of condensin promotes its association with histone H2A and H2A.Z, which we identify as conserved chromatin 'receptors' of condensin. Condensin phosphorylation and its deposition onto chromosome arms reach a peak during anaphase, when Aurora B kinase relocates from centromeres to the spindle midzone, where the separating chromosome arms are positioned. Our results elucidate the molecular basis for the spatiotemporal regulation of mitotic chromosome architecture, which is crucial for chromosome partitioning. PMID- 21633355 TI - A current filamentation mechanism for breaking magnetic field lines during reconnection. AB - During magnetic reconnection, the field lines must break and reconnect to release the energy that drives solar and stellar flares and other explosive events in space and in the laboratory. Exactly how this happens has been unclear, because dissipation is needed to break magnetic field lines and classical collisions are typically weak. Ion-electron drag arising from turbulence, dubbed 'anomalous resistivity', and thermal momentum transport are two mechanisms that have been widely invoked. Measurements of enhanced turbulence near reconnection sites in space and in the laboratory support the anomalous resistivity idea but there has been no demonstration from measurements that this turbulence produces the necessary enhanced drag. Here we report computer simulations that show that neither of the two previously favoured mechanisms controls how magnetic field lines reconnect in the plasmas of greatest interest, those in which the magnetic field dominates the energy budget. Rather, we find that when the current layers that form during magnetic reconnection become too intense, they disintegrate and spread into a complex web of filaments that causes the rate of reconnection to increase abruptly. This filamentary web can be explored in the laboratory or in space with satellites that can measure the resulting electromagnetic turbulence. PMID- 21633356 TI - Mapping in vivo protein-RNA interactions at single-nucleotide resolution from HITS-CLIP data. AB - Mammalian RNA complexity is regulated through interactions of RNA-binding proteins (RBPs) with their target transcripts. High-throughput sequencing together with UV-crosslinking and immunoprecipitation (HITS-CLIP) is able to globally map RBP-binding footprint regions at a resolution of ~30-60 nucleotides. Here we describe a systematic way to analyze HITS-CLIP data to identify exact crosslink sites, and thereby determine protein-RNA interactions at single nucleotide resolution. We found that reverse transcriptase used in CLIP frequently skips the crosslinked amino-acid-RNA adduct, resulting in a nucleotide deletion. Genome-wide analysis of these crosslinking-induced mutation sites (CIMS) in HITS-CLIP data for Nova and Argonaute (Ago) proteins in mouse brain tissue revealed deletions in ~8-20% of mRNA tags, which mapped to Nova and Ago binding sites on mRNA or miRNA. CIMS analysis provides a general and more precise means of mapping protein-RNA interactions than currently available methods and insight into the biochemical properties of such interactions in living tissues. PMID- 21633357 TI - Role of Galpha(12)- and Galpha(13)-protein subunit linkage of D(3) dopamine receptors in the natriuretic effect of D(3) dopamine receptor in kidney. AB - The D(3) dopamine receptor is the major D(2)-like receptor that regulates sodium transport in the renal proximal tubule (RPT) and helps maintain blood pressure in the normal range. In Wistar-Kyoto (WKY) rats chronically fed high-salt diet, the intrarenal arterial infusion of a D(3) receptor agonist, PD128907, increased absolute and fractional sodium excretion. We have reported that Galpha(12) and Galpha(13), which participate in the signal transduction of the D(5) receptor, are expressed in RPTs. As the D(3) receptor is also expressed in RPTs, we hypothesized that it may also interact with Galpha(12)/Galpha(13) in RPTs from WKY rats. There were co-localization and co-immunoprecipitation of D(3) receptor and Galpha(12)/Galpha(13) in renal brush border membranes (BBMs) and RPT cells. The intrarenal infusion of PD128907 (1 MUg kg(-1) min(-1)) that increased sodium excretion also increased the co-immunoprecipitations of D(3)/Galpha(12) and D(3)/Galpha(13) in renal BBMs; their co-immunoprecipitation was confirmed in RPT cells. As Galpha(12) and Galpha(13) increase sodium pump and transporter activity (for example, Na(+)-K(+)-ATPase, NHE3), an increased association of D(3) receptors with Galpha(12)/Galpha(13) receptors after D(3) receptor activation may be a mechanism to prevent Galpha(12)/Galpha(13)-mediated stimulation of sodium transport (and thus enhance natriuresis). We conclude that a D(3) receptor interaction with Galpha(12)/Galpha(13) that increases sodium excretion may have a role in the regulation of blood pressure. PMID- 21633358 TI - Pressure induces intracellular calcium changes in juxtaglomerular cells in perfused afferent arterioles. AB - Calcium (Ca(2+)) has an important role in nearly all types of cellular secretion, with a particularly novel role in the juxtaglomerular (JG) cells in the kidney. In JG cells, Ca(2+) inhibits renin secretion, which is a major regulator of blood pressure and renal hemodynamics. However, whether alterations in afferent arteriolar (Af-Art) pressure change intracellular Ca(2+) concentration ([Ca(2+)](i)) in JG cells and whether [Ca(2+)](i) comes from extracellular or intracellular sources remains unknown. We hypothesize that increases in perfusion pressure in the Af-Art result in elevations in [Ca(2+)](i) in JG cells. We isolated and perfused Af-Art of C57BL6 mice and measured changes in [Ca(2+)](i) in JG cells in response to perfusion pressure changes. The JG cells' [Ca(2+)](i) was 93.3+/-2.2 nM at 60 mm Hg perfusion pressure and increased to 111.3+/-13.4, 119.6+/-7.3, 130.3+/-2.9 and 140.8+/-12.1 nM at 80, 100, 120 and 140 mm Hg, respectively. At 120 mm Hg, increases in [Ca(2+)](i) were reduced in mice receiving the following treatments: (1) the mechanosensitive cation channel blocker, gadolinium (94.6+/-7.5 nM); (2) L-type calcium channel blocker, nifedipine (105.8+/-7.5 nM); and (3) calcium-free solution plus ethylene glycol tetraacetic acid (96.0+/-5.8 nM). Meanwhile, the phospholipase C inhibitor, inositol triphosphate receptor inhibitor, T-type calcium channel blocker, N-type calcium channel blocker and Ca(2+)-ATPase inhibitor did not influence changes in [Ca(2+)](i) in JG cells. In summary, JG cell [Ca(2+)](i) rise as perfusion pressure increases; furthermore, the calcium comes from extracellular sources, specifically mechanosensitive cation channels and L-type calcium channels. PMID- 21633359 TI - Rho-kinase/nuclear factor-kappabeta/angiotensinogen axis in angiotensin II induced renal injury. PMID- 21633360 TI - Maternal age effect on the development of Prader-Willi syndrome resulting from upd(15)mat through meiosis 1 errors. AB - Prader-Willi syndrome (PWS) is primarily caused by deletions involving the paternally derived imprinted region at chromosome 15q11.2-q13 and maternal uniparental disomy 15 (upd(15)mat). The underlying mechanisms for upd(15)mat include trisomy rescue (TR), gamete complementation (GC), monosomy rescue and post-fertilization mitotic error, and TR/GC is mediated by non-disjunction at maternal meiosis 1 (M1) or meiosis 2 (M2). Of these factors involved in the development of upd(15)mat, M1 non-disjunction is a maternal age-dependent phenomenon. We studied 117 Japanese patients with PWS and identified deletions in 84 patients (Deletion group) and TR/GC type upd(15)mat through M1 non-disjunction in 15 patients (TR/GC (M1) group), together with other types of abnormalities. Maternal age was significantly higher in TR/GC (M1) group than in Deletion group (median (range), 37 (35-45) versus 30 (19-42); P=1.0 * 10(-7)). Furthermore, delayed childbearing age became obvious since the year 2003 in Japan, and relative frequency of TR/GC (M1) group was significantly larger in patients born since the year 2003 than in those born until the year 2002. The results imply that the advanced maternal age at childbirth is a predisposing factor for the development of upd(15)mat because of increased M1 errors. PMID- 21633361 TI - Germ-line sequence variants of PTEN do not have an important role in hereditary and non-hereditary prostate cancer susceptibility. AB - PTEN (phosphatase and tensin homolog deleted on chromosome 10) functions as a major tumor suppressor gene and is frequently deleted in different types of tumors including prostate cancer (PCa). It was hypothesized that germ-line genetic changes of PTEN affect susceptibility to PCa. Both common (with a minor allele frequency ?5%) and rare (with a minor allele frequency <5%) germ-line variants of PTEN were comprehensively evaluated. A total of 15 germ-line variants were identified by re-sequencing the PTEN gene, including 5' untranslated region, all nine exons, exon-intron junctions and 3' untranslated region, in 188 probands of hereditary prostate cancer (HPC) families recruited from Johns Hopkins Hospital. Two microsatellite markers surrounding PTEN were used to test the co segregation of 10 rare variants, which may give rise to highly penetrance in HPC. Two common single nucleotide polymorphisms (SNPs) were evaluated in the 188 HPC families using a family-based association study approach. To study low penetrant SNPs in PCa susceptibility, 33 SNPs covering PTEN were selected from the whole genome-wide association studies (GWAS) from our available case-control studies in Sweden (Cancer of the Prostate in Sweden (CAPS)) and the publicly available cancer genetic markers of susceptibility (CGEMS) study. Germ-line copy-number variations (CNVs) in PTEN were assessed in CAPS. Co-segregation of germ-line variants and PCa was not observed among HPC families and no significant differences in the allele frequencies were observed in sporadic cases and controls, aggressive and non-aggressive PCa (P>0.05). These results suggest that germ-line variants in PTEN do not have an important role in PCa susceptibility. PMID- 21633362 TI - Loss-of-function mutation of collybistin is responsible for X-linked mental retardation associated with epilepsy. AB - Microarray-based comparative genomic hybridization analysis identified a 737-kb microdeletion of Xq11.1, including the cell division cycle 42 guanine nucleotide exchange factor (GEF)-9 gene (ARHGEF9), encoding collybistin, which has a pivotal role in formation of postsynaptic glycine and gamma-aminobutyric acid receptor clusters, in a male patient with severe mental retardation and epilepsy. No overlapping deletion with this was identified in the database of genomic copy number variations. A cohort study of ARHGEF9 nucleotide sequence identified a nonsense mutation in another male patient with severe mental retardation and epilepsy. This mutation affects one of the three transcript variants of ARHGEF9, which was confirmed to be expressed in the brain by reverse transcription-PCR. Although this nonsense mutation was shared with the patient's mother, it was not observed in 100 normal individuals. Both male patients suffered epileptic seizures after 1 year of age. Brain magnetic resonance imaging revealed mild frontal atrophy in the first patient and right frontal polymicrogyria in the second patient. Three previously reported mutations of ARHGEF9 consisted of a missense mutation in a male patient with hyperekplexia and two chromosomal disruptions in two female patients. The common phenotypic effects of all ARHGEF9 mutations were mental retardation and epilepsy. Therefore, ARHGEF9 is likely to be responsible for syndromic X-linked mental retardation associated with epilepsy. PMID- 21633363 TI - A population-based study of copy number variants and regions of homozygosity in healthy Swedish individuals. AB - The abundance of copy number variants (CNVs) and regions of homozygosity (ROHs) have been well documented in previous studies. In addition, their roles in complex diseases and traits have since been increasingly appreciated. However, only a limited amount of CNV and ROH data is currently available for the Swedish population. We conducted a population-based study to detect and characterize CNVs and ROHs in 87 randomly selected healthy Swedish individuals using the Affymetrix SNP Array 6.0. More than 600 CNV loci were detected in the population using two different CNV-detection algorithms (PennCNV and Birdsuite). A total of 196 loci were consistently identified by both algorithms, suggesting their reliability. Numerous disease-associated and pharmacogenetics-related genes were found to be overlapping with common CNV loci such as CFHR1/R3, LCE3B/3C, UGT2B17 and GSTT1. Correlation analysis between copy number polymorphisms (CNPs) and genome-wide association studies-identified single-nucleotide polymorphisms also indicates the potential roles of several CNPs as causal variants for diseases and traits such as body mass index, Crohn's disease and multiple sclerosis. In addition, we also identified a total of 14 815 ROHs ?500 kb or 2814 ROHs ?1M in the Swedish individuals with an average of 170 and 32 regions detected per individual respectively. Approximately 141 Mb or 4.92% of the genome is homozygous in each individual of the Swedish population. This is the first population-based study to investigate the population characteristics of CNVs and ROHs in the Swedish population. This study found many CNV loci that warrant further investigation, and also highlighted the abundance and importance of investigating ROHs for their associations with complex diseases and traits. PMID- 21633364 TI - Congenital factor XIII deficiency: a commentary on 'Homozygous intronic mutation leading to inefficient transcription combined with a novel frame-shift mutation in F13A1 gene causes FXIII deficiency'. PMID- 21633366 TI - A Commentary on Identification of an autosomal dominant locus for intracranial aneurysm through a model-based family collection in a geographically limited area. PMID- 21633365 TI - Molecular and clinical studies of X-linked deafness among Pakistani families. AB - There are 68 sex-linked syndromes that include hearing loss as one feature and five sex-linked nonsyndromic deafness loci listed in the OMIM database. The possibility of additional such sex-linked loci was explored by ascertaining three unrelated Pakistani families (PKDF536, PKDF1132 and PKDF740) segregating X-linked recessive deafness. Sequence analysis of POU3F4 (DFN3) in affected members of families PKDF536 and PKDF1132 revealed two novel nonsense mutations, p.Q136X and p.W114X, respectively. Family PKDF740 is segregating congenital blindness, mild to-profound progressive hearing loss that is characteristic of Norrie disease (MIM#310600). Sequence analysis of NDP among affected members of this family revealed a novel single nucleotide deletion c.49delG causing a frameshift and premature truncation (p.V17fsX1) of the encoded protein. These mutations were not found in 150 normal DNA samples. Identification of pathogenic alleles causing X linked recessive deafness will improve molecular diagnosis, genetic counseling and molecular epidemiology of hearing loss among Pakistanis. PMID- 21633367 TI - The association between Peyronie's and Dupuytren's disease. AB - Peyronie's disease (PD) is known to be associated with Dupuytren's disease (DD) since 1828. The aim of this study was to investigate the coexistence of DD in a consecutive series of patients with PD and their clinical characteristics. From January 1988 to December 2009 all patients, presenting at our outpatient urological clinic, with PD were also examined for DD. The sample consisted of 415 male subjects with PD, 89 (22.1%) also had DD. A total of 28 men (6.7%) reported to have one or more first or second degree relatives with DD. PMID- 21633368 TI - HIF-1alpha in epidermis: oxygen sensing, cutaneous angiogenesis, cancer, and non cancer disorders. AB - Besides lung, postnatal human epidermis is the only epithelium in direct contact with atmospheric oxygen. Skin epidermal oxygenation occurs mostly through atmospheric oxygen rather than tissue vasculature, resulting in a mildly hypoxic microenvironment that favors increased expression of hypoxia-inducible factor 1alpha (HIF-1alpha). Considering the wide spectrum of biological processes, such as angiogenesis, inflammation, bioenergetics, proliferation, motility, and apoptosis, that are regulated by this transcription factor, its high expression level in the epidermis might be important to HIF-1alpha in skin physiology and pathophysiology. Here, we review the role of HIF-1alpha in cutaneous angiogenesis, skin tumorigenesis, and several skin disorders. PMID- 21633369 TI - Protective effect of Kit signaling for melanocyte stem cells against radiation induced genotoxic stress. AB - Radiation-induced hair graying is caused by irreversible defects in the self renewal and/or development of follicular melanocyte stem cells in the hair follicles. Kit signaling is an essential growth and differentiation signaling pathway for various cell lineages including melanocytes, and its radioprotective effects have been shown in hematopoietic cells. However, it is uncertain whether Kit signaling exerts a radioprotective effect for melanocytes. In this study, we found that various loss-of-function mutations of Kit facilitate radiation-induced hair graying. In contrast, transgenic mice expressing the ligand for Kit (Kitl) in the epidermis have significantly reduced levels of radiation-induced hair graying. The X-ray doses used did not show a systemic lethal effect, indicating that the in vivo radiosensitivity of Kit mutants is mainly caused by the damaged melanocyte stem cell population. X-ray-damaged melanocyte stem cells seemed to take the fate of ectopically pigmented melanocytes in the bulge regions of hair follicles in vivo. Endothelin 3, another growth and differentiation factor for melanocytes, showed a lesser radioprotective effect compared with Kitl. These results indicate the prevention of radiation-induced hair graying by Kit signaling. PMID- 21633370 TI - Migration of immunocytes across the basement membrane in skin: the role of basement membrane pores. PMID- 21633371 TI - Efficacy of combined peroxisome proliferator-activated receptor-alpha ligand and glucocorticoid therapy in a murine model of atopic dermatitis. AB - Although topical glucocorticoids (GCs) show potent anti-inflammatory activity in inflamed skin, they can also exert numerous harmful effects on epidermal structure and function. In contrast, topical applications of ligands of peroxisome proliferator-activated receptor-alpha (PPARalpha) not only reduce inflammation but also improve cutaneous barrier homeostasis. Therefore, we examined whether sequential topical GCs followed by topical Wy14643 (a ligand of PPARalpha) might be more effective than either alone for atopic dermatitis (AD) in a hapten (oxazolone (Ox))-induced murine model with multiple features of AD (Ox-AD). Despite expected anti-inflammatory benefits, topical GC alone induced (i) epidermal thinning; (ii) reduced expression of involucrin, loricrin, and filaggrin; and (iii) allowed outside-to-inside penetration of an epicutaneous tracer. Although Wy14643 alone yielded significant therapeutic benefits in mice with mild or moderate Ox-AD, it was less effective in severe Ox-AD. Yet, topical application of Wy14643 after GC was not only significantly effective comparable with GC alone, but it also prevented GC-induced structural and functional abnormalities in permeability barrier homeostasis. Moreover, rebound flares were largely absent after sequential treatment with GC and Wy14643. Together, these results show that GC and PPARalpha ligand therapy together is not only effective but also prevents development of GC-induced side effects, including rebound flares, in murine AD. PMID- 21633372 TI - AKT has an anti-apoptotic role in ABCA12-deficient keratinocytes. PMID- 21633373 TI - FLI1 polymorphism affects susceptibility to cutaneous leishmaniasis in Brazil. AB - Mapping murine genes controlling cutaneous leishmaniasis (CL) identified Fli1 as a candidate influencing resistance to L. major and enhanced wound healing. We examine FLI1 as a gene controlling CL and mucosal leishmaniasis (ML) caused by L. braziliensis in humans. Intron 1 single nucleotide polymorphisms tagging promoter and enhancer elements were analysed in 168 nuclear families (250 CL; 87 ML cases) and replicated in 157 families (402 CL; 39 ML cases). Robust case-pseudocontrol logistic regression analysis showed association between allele C (odds ratio (OR) 1.65; 95% confidence interval 1.18-2.29; P=0.003) of FLI1_rs7930515 and CL in the primary sample that was confirmed (OR 1.60; 95% confidence interval 1.10-2.33; P=0.014) in the replication set (combined P=1.8 * 10(-4)). FLI1_rs7930515 is in linkage disequilibrium with the functional GAn microsatellite in the proximal promoter. Haplotype associations extended across the enhancer, which was not polymorphic. ML associated with inverse haplotypes compared with CL. Wound healing is therefore important in CL, providing potential for therapies modulating FLI1. PMID- 21633374 TI - When inordinate tissue growth is beneficial: improving silk production by increasing silk gland size. PMID- 21633377 TI - The combined impact of 12 common variants on hypertension in Japanese men, considering GWAS results. AB - Genome-wide association studies have identified several polymorphisms that appear to be on hypertension-susceptible regions. We performed the current replication study in order to evaluate the association of these loci with hypertension in healthy Japanese males and then examined the combined effect of 12 independent variants. Overall, 735 Japanese men from two independent cohorts were recruited. Association with hypertension was assessed in 16 polymorphisms on 12 genes and 12 were chosen to evaluate the combined impact. Polymorphisms on the COMT, ATP2B1, CYP11A1 and the CSK genes were confirmed to be associated with hypertension and blood pressure (BP). Current findings also replicated previous results for the CYP11B2 and PTGIS genes. Although there were no significant associations found for other variants, our results suggested there was a combined impact for 12 loci. Individuals carrying more risk alleles had a higher risk of hypertension (P for the slope=0.002). Blood pressures also increased in conjunction with an increasing risk allele score (P for trend=7.84 * 10(-6) and 1.85 * 10(-5) for SBP and DBP, respectively). Our results confirmed the associations between hypertension or blood pressure and four gene variants. We also found a significant combined effect of the 12 gene loci. PMID- 21633378 TI - Cardiovascular risk factor(s) prevalence in Greek hypertensives. Effect of gender and age. AB - The aim of this study was to determine cardiovascular (CV) risk factors (RFs) and target organ damage clustering in 21280 Greek hypertensives stratified by gender and age. Glycemic and lipid profile were determined, left ventricular mass index, estimated gromerular filtration rate (eGFR), 10-years CV risk according to Framingham risk score (FRS) and HeartScore (HS) were calculated. Only 10.2% of patients had no concomitant RFs, 53.1% had one (48.8% dyslipidemia, 3.4% smoking, 0.9% diabetes), 32.9% had two (26% dyslipidemia and smoking, 6.6% dyslipidemia and diabetes, 0.3% smoking and diabetes) and 3.7% had all four traditional RFs. Obesity was present in 30%, metabolic syndrome in 38%, low eGFR in 24% and left ventricular hypertrophy in 49%. Mean FRS risk was 35% for males, 24.1% for females whereas in high risk (>20%) were 68.7 and 50.7%, respectively (P<0.0001). Mean HS risk was 8.4% for males, 6.2% for females whereas in high risk (>5%) were 48.6 and 36.2%, respectively (P<0.0001). Age was correlated to pulse pressure, eGFR, left ventricular mass index and CV risk (P<0.0001). Ageing increased the risk difference between genders for total (P=0.001) but not for fatal events (P=nonsignificant). In conclusion, as RFs cluster in hypertensives, CV risk calculation should guide treatment decisions. PMID- 21633379 TI - Geographic variation in left ventricular mass and mass index: a systematic review. AB - Left ventricular (LV) hypertrophy, defined as an abnormal increase in LV mass (LVM), is an important prognostic indicator and therapeutic target. LVM is often divided by body surface area to derive indexed mass; however, this does not correctly identify pathological LV hypertrophy in all people, especially when body composition is altered, or in different ethnic groups. We evaluated published ranges of echocardiographic LVM in healthy adult populations from different countries, excluding control groups, and compared them with the American Society of Echocardiography reference ranges. A total of 33 studies met the inclusion criteria. In men and women, there was wide variation in the ranges of LVM with a tendency for the upper limit to increase geographically westward; this variation remained for indexed mass. Several ranges fell outside the upper reference limits: in men, 13 of the mass ranges and 16 of indexed mass; and in women, 8 mass and 16 indexed mass. This review has shown that current guidelines may need revision as some published series suggest that greater LV mass should be considered normal. This may be explained by ethnic differences and supports the need for widely applicable and ethnically diverse reference ranges to be established. PMID- 21633380 TI - Cognitive and neurobiological mechanisms of alcohol-related aggression. AB - Alcohol-related violence is a serious and common social problem. Moreover, violent behaviour is much more common in alcohol-dependent individuals. Animal experiments and human studies have provided insights into the acute effect of alcohol on aggressive behaviour and into common factors underlying acute and chronic alcohol intake and aggression. These studies have shown that environmental factors, such as early-life stress, interact with genetic variations in serotonin-related genes that affect serotonergic and GABAergic neurotransmission. This leads to increased amygdala activity and impaired prefrontal function that, together, predispose to both increased alcohol intake and impulsive aggression. In addition, acute and chronic alcohol intake can further impair executive control and thereby facilitate aggressive behaviour. PMID- 21633382 TI - Neurodegenerative disease: directing amyloid-beta deposition. PMID- 21633381 TI - Differentiating the rapid actions of cocaine. AB - The subjective effects of intravenous cocaine are felt almost immediately, and this immediacy plays an important part in the drug's rewarding impact. The primary rewarding effect of cocaine involves blockade of dopamine reuptake; however, the onset of this action is too late to account for the drug's initial effects. Recent studies suggest that cocaine-predictive cues--including peripheral interoceptive cues generated by cocaine itself--come to cause more direct and earlier reward signalling by activating excitatory inputs to the dopamine system. The conditioned activation of the dopamine system by cocaine predictive cues offers a new target for potential addiction therapies. PMID- 21633383 TI - Auditory processing: sounding out consciousness. PMID- 21633385 TI - Stem cells: cNeoblasts keep their options open. PMID- 21633384 TI - A blueprint for kinetochores - new insights into the molecular mechanics of cell division. AB - Kinetochores are large proteinaceous complexes that physically link centromeric DNA to the plus ends of spindle microtubules. Stable kinetochore-microtubule attachments are a prerequisite for the accurate and efficient distribution of genetic material over multiple generations. In the past decade, concerted research has resulted in the identification of the individual kinetochore building blocks, the characterization of critical microtubule-interacting components, such as the NDC80 complex, and the development of an approximate model of the architecture of this sophisticated biological machine. PMID- 21633386 TI - DNA repair: BRCA2 gets protective at forks. PMID- 21633387 TI - Cubism and the cell cycle: the many faces of the APC/C. AB - One does not often look to analytic cubism for insights into the control of the cell cycle, but Pablo Picasso beautifully encapsulated the fundamentals when he said that "every act of creation is, first of all, an act of destruction". The rapid destruction of specific cell cycle regulators at just the right moment in the cell cycle ensures that daughter cells receive an equal and identical set of chromosomes from their mother and that DNA replication always follows mitosis. Remarkably, one protein complex is responsible for this surgical precision, the APC/C (anaphase-promoting complex, also known as the cyclosome). The APC/C is tightly regulated by its co-activators and by the spindle assembly checkpoint. PMID- 21633388 TI - Differentiation of hematopoietic stem cell and myeloid populations by ATP is modulated by cytokines. AB - Extracellular nucleotides are emerging as important regulators of inflammation, cell proliferation and differentiation in a variety of tissues, including the hematopoietic system. In this study, the role of ATP was investigated during murine hematopoiesis. ATP was able to reduce the percentage of hematopoietic stem cells (HSCs), common myeloid progenitors and granulocyte-macrophage progenitors (GMPs), whereas differentiation into megakaryocyte-erythroid progenitors was not affected. In addition, in vivo administration of ATP to mice reduced the number of GMPs, but increased the number of Gr-1(+)Mac-1(+) myeloid cells. ATP also induced an increased proliferation rate and reduced Notch expression in HSCs and impaired HSC-mediated bone marrow reconstitution in sublethally irradiated mice. Moreover, the effects elicited by ATP were inhibited by suramin, a P2 receptor antagonist, and BAPTA, an intracellular Ca(2+) chelator. We further investigated whether the presence of cytokines might modulate the observed ATP-induced differentiation. Treatment of cells with cytokines (stem cell factor, interleukin 3 and granulocyte-monocyte colony stimulator factor) before ATP stimulation led to reduced ATP-dependent differentiation in long-term bone marrow cultures, thereby restoring the ability of HSCs to reconstitute hematopoiesis. Thus, our data suggest that ATP induces the differentiation of murine HSCs into the myeloid lineage and that this effect can be modulated by cytokines. PMID- 21633389 TI - Utilizing mitochondrial events as biomarkers for imaging apoptosis. AB - Cells undergoing apoptosis show a plethora of time-dependent changes. The available tools for imaging apoptosis in live cells rely either on the detection of the activity of caspases, or on the visualization of exposure of phosphatidyl serine in the outer leaflet of the cell membrane. We report here a novel method for the detection of mitochondrial events during apoptosis, namely translocation of Bax to mitochondria and release of cytochrome c (Cyt c) using bimolecular fluorescence complementation. Expression of split yellow fluorescent protein (YFP) fragments fused to Bax and Cyt c, resulted in robust induction of YFP fluorescence at the mitochondria of apoptotic cells with very low background. In vivo expression of split YFP protein fragments in liver hepatocytes and intra vital imaging of subcutaneous tumor showed elevated YFP fluorescence upon apoptosis induction. Thus, YFP complementation could be applied for high throughput screening and in vivo molecular imaging of mitochondrial events during apoptosis. PMID- 21633390 TI - Astrocytes are important mediators of Abeta-induced neurotoxicity and tau phosphorylation in primary culture. AB - Alzheimer's disease (AD) is pathologically characterised by the age-dependent deposition of beta-amyloid (Abeta) in senile plaques, intraneuronal accumulation of tau as neurofibrillary tangles, synaptic dysfunction and neuronal death. Neuroinflammation, typified by the accumulation of activated microglia and reactive astrocytes, is believed to modulate the development and/or progression of AD. We have used primary rat neuronal, astrocytic and mixed cortical cultures to investigate the contribution of astrocyte-mediated inflammatory responses during Abeta-induced neuronal loss. We report that the presence of small numbers of astrocytes exacerbate Abeta-induced neuronal death, caspase-3 activation and the production of caspase-3-cleaved tau. Furthermore, we show that astrocytes are essential for the Abeta-induced tau phosphorylation observed in primary neurons. The release of soluble inflammatory factor(s) from astrocytes accompanies these events, and inhibition of astrocyte activation with the anti-inflammatory agent, minocycline, reduces astrocytic inflammatory responses and the associated neuronal loss. Abeta-induced increases in caspase-3 activation and the production of caspase-3-truncated tau species in neurons were reduced when the astrocytic response was attenuated with minocycline. Taken together, these results show that astrocytes are important mediators of the neurotoxic events downstream of elevated Abeta in models of AD, and suggest that mechanisms underlying pro inflammatory cytokine release might be an important target for therapy. PMID- 21633391 TI - Salinomycin induces calpain and cytochrome c-mediated neuronal cell death. AB - Salinomycin is a polyether antibiotic with properties of an ionophore, which is commonly used as cocciodiostatic drug and has been shown to be highly effective in the elimination of cancer stem cells (CSCs) both in vitro and in vivo. One important caveat for the potential clinical application of salinomycin is its marked neural and muscular toxicity. In the present study we show that salinomycin in concentrations effective against CSCs exerts profound toxicity towards both dorsal root ganglia as well as Schwann cells. This toxic effect is mediated by elevated cytosolic Na(+) concentrations, which in turn cause an increase of cytosolic Ca(2+) by means of Na(+)/Ca(2+) exchangers (NCXs) in the plasma membrane as well as the mitochondria. Elevated Ca(2+) then leads to calpain activation, which triggers caspase-dependent apoptosis involving caspases 12, 9 and 3. In addition, cytochrome c released from depolarized mitochondria directly activates caspase 9. Combined inhibition of calpain and the mitochondrial NCXs resulted in significantly decreased cytotoxicity and was comparable to caspase 3 inhibition. These findings improve our understanding of mechanisms involved in the pathogenesis of peripheral neuropathy and are important to devise strategies for the prevention of neurotoxic side effects induced by salinomycin. PMID- 21633392 TI - Developing therapeutic microRNAs for cancer. AB - Despite substantial progress in understanding the cancer-signaling network, effective therapies remain scarce due to insufficient disruption of oncogenic pathways, drug resistance and drug-induced toxicity. This complexity of cancer defines an urgent goal for researchers and clinicians to develop novel therapeutic strategies. The discovery of microRNAs (miRNAs) provides new hope for accomplishing this task. Supported by solid evidence for a critical role in cancer and bolstered by a unique mechanism of action, miRNAs are likely to yield a new class of targeted therapeutics. In contrast to current cancer medicines, miRNA-based therapies function by subtle repression of gene expression on a yet large number of oncogenic factors and are, therefore, anticipated to be highly efficacious. After the completion of target validation for several candidates, the development of therapeutic miRNAs is now moving to a new stage that involves pharmacological drug delivery, preclinical toxicology and regulatory guidelines. PMID- 21633393 TI - Targeted suicide gene therapy for glioma using human embryonic stem cell-derived neural stem cells genetically modified by baculoviral vectors. AB - Tumor-tropic neural stem cells (NSCs) can be used in the Trojan horse approach as cellular vehicles for targeted delivery of therapeutic agents to distant tumor sites. To realize this cancer therapy potential, it is important to have a renewable source to generate large quantities of uniform human NSCs. Here, we reported that NSCs derived from HES1 human embryonic stem cell line were capable of migrating into intracranial glioma xenografts after systemic injection or after intracranial injection at a site distant from the tumor. To test whether the HES1-derived NSCs can be used for cancer gene therapy, we used a baculoviral vector to introduce the herpes simplex virus thymidine kinase suicide gene into the cells and demonstrated that baculovirus-mediated transgene expression may last for at least 3 weeks in NSCs. After being injected into the cerebral hemisphere opposite the tumor site and in the presence of ganciclovir, NSCs expressing the suicide gene were able to inhibit the growth of human glioma xenografts and prolong survival of tumor-bearing mice. Our findings suggest that human embryonic stem cells could potentially serve as a clinically viable source for production of cellular vehicles suitable for targeted anticancer gene therapy. PMID- 21633394 TI - Supercoiled Minivector DNA resists shear forces associated with gene therapy delivery. AB - Supercoiled DNAs varying from 281 to 5302 bp were subjected to shear forces generated by aerosolization or sonication. DNA shearing strongly correlated with length. Typical sized plasmids (>= 3000 bp) degraded rapidly. DNAs 2000-3000 bp persisted ~10 min. Even in the absence of condensing agents, supercoiled DNA <1200 bp survived nebulization, and increased forces of sonication were necessary to shear it. Circular vectors were considerably more resistant to shearing than linear vectors of the same length. DNA supercoiling afforded additional protection. These results show the potential of shear-resistant Minivector DNAs to overcome one of the major challenges associated with gene therapy delivery. PMID- 21633395 TI - The diversity of cyanomyovirus populations along a North-South Atlantic Ocean transect. AB - Viruses that infect the marine cyanobacterium Prochlorococcus have the potential to impact the growth, productivity, diversity and abundance of their hosts. In this study, changes in the microdiversity of cyanomyoviruses were investigated in 10 environmental samples taken along a North-South Atlantic Ocean transect using a myoviral-specific PCR-sequencing approach. Phylogenetic analyses of 630 viral g20 clones from this study, with 786 published g20 sequences, revealed that myoviral populations in the Atlantic Ocean had higher diversity than previously reported, with several novel putative g20 clades. Some of these clades were detected throughout the Atlantic Ocean. Multivariate statistical analyses did not reveal any significant correlations between myoviral diversity and environmental parameters, although myoviral diversity appeared to be lowest in samples collected from the north and south of the transect where Prochlorococcus diversity was also lowest. The results were correlated to the abundance and diversity of the co-occurring Prochlorococcus and Synechococcus populations, but revealed no significant correlations to either of the two potential host genera. This study provides evidence that cyanophages have extremely high and variable diversity and are distributed over large areas of the Atlantic Ocean. PMID- 21633396 TI - Prehypertension is associated with impaired nitric oxide-mediated endothelium dependent vasodilation in sedentary adults. AB - BACKGROUND: Endothelial vasodilator dysfunction contributes to the development of hypertension (blood pressure (BP) >= 140/90 mm Hg) and cardiovascular disease (CVD). Prehypertension (BP 120-139/80-89 mm Hg) has recently been identified as an independent risk factor for hypertension and CVD. It is currently unclear whether BP in the prehypertensive range is associated with endothelial vasodilator dysfunction. We tested the hypothesis that BP in the prehypertensive range, independent of other cardiovascular risk factors, is associated with impaired nitric oxide (NO)-mediated endothelium-dependent vasodilation. METHODS: Forearm blood flow (FBF) responses to intra-arterial acetylcholine (ACh; 8.0-32.0 ug/100 ml tissue/min) and sodium nitroprusside (SNP; 1.0-4.0 ug/100 ml tissue/min) were measured in 20 normotensive (age: 56 +/- 1 years; BP: 110/70 +/- 1/2 mm Hg) and 20 prehypertensive (56 +/- 2 years; 128/79 +/- 2/2 mm Hg) adults. In addition, FBF responses to ACh were determined in the absence and presence of the endothelial NO synthase inhibitor N(G)-monomethyl-L-arginine (L-NMMA) (5 mg/min). RESULTS: FBF responses to ACh were significantly lower (~30%) in prehypertensive (from 4.2 +/- 0.3 to 11.4 +/- 0.7 ml/100 ml tissue/min) compared with normotensive (from 4.6 +/- 0.2 to 14.5 +/- 0.7 ml/100 ml tissue/min) adults. There were no group differences in FBF responses to SNP. Co-infusion of L-NMMA significantly reduced the FBF response to ACh in the normotensive (~30%; P <0.05) but not the prehypertensive adults. CONCLUSIONS: Prehypertension is associated with impaired NO-mediated endothelium-dependent vasodilation. The endothelial vasodilator dysfunction that characterizes hypertension is present at BP levels in the prehypertensive range and may contribute to the increased risk of hypertension and CVD in this population. PMID- 21633397 TI - Blood pressure dipping: ethnicity, sleep quality, and sympathetic nervous system activity. AB - BACKGROUND: Blunted blood pressure (BP) dipping is an established predictor of adverse cardiovascular outcomes. Although blunted BP dipping is more common in African Americans than whites, the factors contributing to this ethnic difference are not well understood. This study examined the relationships of BP dipping to ethnicity, body mass index (BMI), sleep quality, and fall in sympathetic nervous system (SNS) activity during the sleep-period. METHODS: On three occasions, 128 participants with untreated high clinic BP (130-159/85-99 mm Hg) underwent assessments of 24-h ambulatory BP (ABP), sleep quality, (evaluated by sleep interview, self-report, actigraphy) and sleep-period fall in sympathetic activity (measured by waking/sleep urinary catecholamine excretion). RESULTS: Compared to whites (n = 72), African Americans (n = 56) exhibited higher sleep-period systolic (SBP) (P = 0.01) and diastolic BP (DBP) (P < 0.001), blunted SBP dipping (P = 0.01), greater BMI (P = 0.049), and poorer sleep quality (P = 0.02). SBP dipping was correlated with BMI (r = -0.32, P < 0.001), sleep quality (r = 0.30, P < 0.001), and sleep-period fall in sympathetic activity (r = 0.30, P < 0.001). Multiple regression analyses indicated that these three factors were independent determinants of sleep-period SBP dipping; ethnic differences in dipping were attenuated when controlling for these factors. CONCLUSIONS: Blunted BP dipping was related to higher BMI, poorer sleep quality, and a lesser decline in sleep period SNS activity. Although African-American ethnicity also was associated with blunted dipping compared to whites in unadjusted analyses, this ethnic difference was diminished when BMI, sleep quality, and sympathetic activity were taken into account. PMID- 21633398 TI - Altered default network activity in obesity. AB - The regulation of energy intake is a complex process involving the integration of homeostatic signals and both internal and external sensory inputs. To better understand the neurobiology of this process and how it may be dysfunctional in obesity, this study examined activity of the brain's "default network" in reduced obese (RO) as compared to lean individuals. The default network is a group of functionally connected brain regions thought to play an important role in internally directed cognitive activity and the interplay between external and internal sensory processing. Functional magnetic resonance imaging was performed in 24 lean and 18 RO individuals in the fasted state after 2 days of eucaloric energy intake and after 2 days of 30% overfeeding in a counterbalanced design. Scanning was performed while subjects passively viewed images of food and nonfood objects. Independent component analysis was used to identify the default network component. In the eucaloric state, greater default network activity was observed in RO compared to lean individuals in the lateral inferior parietal and posterior cingulate cortices. Activity was positively correlated with appetite. Overfeeding resulted in increased default network activity in lean but not RO individuals. These findings suggest that the function of the default network, a major contributor to intrinsic neuronal activity, is altered in obesity and/or obese prone individuals. Future studies of the network's function and its relationship to other brain networks may improve our understanding of the mechanisms and treatment of obesity. PMID- 21633399 TI - Metabolic syndrome and acute hyperglycemia are associated with endoplasmic reticulum stress in human mononuclear cells. AB - Endoplasmic reticulum (ER) stress and the activation of the unfolded protein response (UPR) have been implicated in a number of complications associated with diabetes mellitus including micro- and macrovascular dysfunction. In this study we examine ER stress levels in blood cells isolated from human subjects with metabolic syndrome and in healthy controls. Total RNA and protein were isolated from leukocytes and the levels of specific ER stress markers were quantified by real-time-PCR and immunoblot analysis. Our results indicate that, compared to healthy controls, individuals with metabolic syndrome have elevated mRNA levels of genes indicative of ER stress; including spliced XBP-1 (sXBP-1), Grp78, and CHOP. Induced ER stress levels correlate with blood glucose but not plasma lipid concentration. Furthermore, in healthy individuals, a standard 75 g oral glucose challenge produced a significant elevation in spliced XBP-1 (1.3 fold), Grp78 (2.0 fold), and calreticulin (3.5 fold) mRNA 60 min post challenge and a significant increase in Grp78 (2.0 fold), calreticulin (2.7 fold) protein levels 2 h postchallenge, relative to fasting levels. The UPR was also activated ex vivo, in human leukocytes cultured in the presence of 15 mmol/l glucose, supporting a specific role for glucose. The oral glucose challenge was associated with a significant increase in the expression of inflammatory cytokines, including interleukin (IL)-1alpha/beta, IL-6, and IL-8, that may result from ER stress. These findings suggest that there is an association between both acute and chronic dysglycemia and ER stress in humans. PMID- 21633400 TI - Obesity and injury-related absenteeism in a population-based firefighter cohort. AB - A consistent relationship has been demonstrated between obesity and absenteeism in the workplace. However, most studies have focused on primarily sedentary occupational groups. Firefighting is a physically demanding profession that involves significant potential for exposure to dangerous situations and strenuous work. No studies to date have evaluated the impact of obesity on risk for absenteeism among firefighters. We examined the cross-sectional association between BMI and obesity and injury-related absenteeism. BMI, body fat percentage (BF%), waist circumference (WC), injury, and injury-related absenteeism were assessed in 478 career male firefighters. One hundred and fifteen firefighters reported an injury in the previous year and the number of days absent from work due to their injury. BMI was an independent predictor of absenteeism due to injury even after adjustment for confounding variables. Firefighters meeting the definition of class II and III obesity had nearly five times (odds ratio (OR) = 4.89; 95% confidence interval (CI) = 3.63-6.58) the number missed work days due to injury when compared to their normal weight counterparts and their elevated risk was greater than firefighters with class I obesity (OR = 2.71; 95% CI = 2.01 3.65) or those who were overweight (OR = 2.55; 95% CI = 1.90-3.41). The attributable per capita costs of class II and III obesity-related absenteeism over the last year were $1,682.90 per firefighter, $254.00 per firefighter for class I obesity, and $74.41 per firefighter for overweight. Our findings suggest that class II and III obesity were associated with substantial attributable costs to employers and our cost estimates probably underestimate the actual financial burden. PMID- 21633401 TI - Age-dependent inverse association between alcohol consumption and obesity in Japanese men. AB - The aim of this study was to determine whether age influences the relationship between alcohol drinking and obesity. Japanese men receiving periodic health checkups (20-70 years old, n = 36,121) were divided into non-, light (<22 g ethanol/day), moderate (>=22 and <44 g ethanol/day), and heavy (>=44 g ethanol/day) drinkers. Relationships between alcohol intake and obesity-related indices were compared among the quartiles of age. BMI was lower in light and moderate drinkers than in nondrinkers, and these differences were more prominent in the 1st and 2nd quartiles of age than in the higher quartiles. In the 1st and 2nd quartiles of age, waist circumference and waist-to-height ratio were significantly smaller and lower, respectively, in light and moderate drinkers than in nondrinkers, and these differences were less prominent in the 3rd quartile and were not found in the 4th quartile. In the 1st and 2nd quartiles of age, odds ratios vs. nondrinkers for large waist circumference were significantly low (P < 0.01) in light drinkers (1st quartile: 0.64 (0.54-0.75); 2nd quartile: 0.69 (0.60-0.80)) and moderate drinkers (1st quartile: 0.69 (0.61-0.78); 2nd quartile: 0.84 (0.76-0.93)), whereas the odds ratio was significantly low (P < 0.05) only in light drinkers in the 3rd quartile (0.84 (0.73-0.97)) and was not significant in any drinker groups of the 4th quartile. The results suggest that alcohol consumption is associated with lower risk of obesity in Japanese men and this association is more prominent in younger men than in older men. PMID- 21633402 TI - Serum gamma-glutamyltransferase: independent predictor of risk of diabetes, hypertension, metabolic syndrome, and coronary disease. AB - Serum gamma-glutamyltransferase (GGT) is associated with oxidative stress and hepatic steatosis. The extent to which its value in determining incident cardiometabolic risk (coronary heart disease (CHD), metabolic syndrome (MetS), hypertension and type 2 diabetes) is independent of obesity needs to be further explored in ethnicities. After appropriate exclusions, a cohort of 1,667 adults of a general population (age 52 +/-11 years) was evaluated prospectively at 4 year's follow-up using partly Cox proportional hazard regressions. GGT activity was measured kinetically, and values were log-transformed for analyses. MetS was identified by Adult Treatment Panel-III criteria modified for male abdominal obesity. Median (interquartile range) GGT activity was 24.9 (17.0; 35.05) U/l in men, 17.0 (12.3; 24.0) U/l in women. In linear regression analysis, while smoking status was not associated, (male) sex, sex-dependent age, alcohol usage, BMI, fasting triglycerides and C-reactive protein (CRP) were significant independent determinants of circulating GGT. Each 1-s.d. increment in (= 0.53 ln GGT) GGT activity significantly predicted in each sex incident hypertension (hazard ratio (HR) 1.20 (95% confidence interval (CI) 1.10; 1.31)), and similarly MetS, after adjustment for age, alcohol usage, smoking status, BMI and menopause. Strongest independent association existed with diabetes (HR 1.3 (95% CI 1.1; 1.5)) whereas GGT activity tended to marginally predict CHD independent of total bilirubin but not of BMI. Higher serum total bilirubin levels were protective against CHD risk in women. We conclude that elevated serum GGT confers, additively to BMI, risk of hypertension, MetS, and type 2 diabetes but only mediates adiposity against CHD risk. PMID- 21633403 TI - Male inclusion in randomized controlled trials of lifestyle weight loss interventions. AB - The prevalence of obesity is similar for men (32.2%) and women (35.5%). It has been assumed that lifestyle weight loss interventions have been developed and tested in predominately female samples, but this has not been systematically investigated. The aim of this review was to investigate total and ethnic male inclusion in randomized controlled trials of lifestyle interventions. PUBMED, MEDLINE, and PSYCHINFO were searched for randomized controlled trials of lifestyle weight loss interventions (N = 244 studies with a total of 95,207 participants) published in the last 10 years (1999-2009). A trial must be in English, included weight loss as an outcome, and tested a dietary, exercise, and/or other behavioral intervention for weight loss. Results revealed samples were on average 27% male vs. 73% female (P < 0.001). Trials recruiting a diseased sample included a larger proportion of males than those not targeting a disease (35% vs. 21%; P < 0.001). About 32% of trials used exclusively female samples, whereas only 5% used exclusively male samples (P < 0.001). No studies in the past 10 years specifically targeted minority males. Ethnic males identified composed 1.8% of total participants in US studies. Only 24% of studies that underrepresented males provided a reason. Males, especially ethnic males, are underrepresented in lifestyle weight loss trials. PMID- 21633405 TI - (-)-Epigallocatechin-3-gallate inhibits pancreatic lipase and reduces body weight gain in high fat-fed obese mice. AB - Tea (Camellia sinensis, Theaceae) has been shown to have obesity preventive effects in laboratory studies. We hypothesized that dietary epigallocatechin-3 gallate (EGCG) could reverse metabolic syndrome in high fat-fed obese C57bl/6J mice, and that these effects were related to inhibition of pancreatic lipase (PL). Following treatment with 0.32% EGCG for 6 weeks, a 44% decrease in body weight (BW) gain in high fat-fed, obese mice (P < 0.01) was observed compared to controls. EGCG treatment increased fecal lipid content by 29.4% (P < 0.05) compared to high fat-fed control, whereas in vitro, EGCG dose-dependently inhibited PL (IC(50) = 7.5 umol/l) in a noncompetitive manner with respect to substrate concentration. (-)-Epicatechin-3-gallate exhibited similar inhibitory activity, whereas the nonester-containing (-)-epigallocatechin did not. In conclusion, EGCG supplementation reduced final BW and BW gain in obese mice, and some of these effects may be due to inhibition of PL by EGCG. PMID- 21633404 TI - Variants at the endocannabinoid receptor CB1 gene (CNR1) and insulin sensitivity, type 2 diabetes, and coronary heart disease. AB - Inhibition of the endocannabinoid receptor CB1 improves insulin sensitivity, lowers glycemia, and slows atherosclerosis. We analyzed whether common variants in the gene encoding CB1, CNR1, are associated with insulin resistance, risk of type 2 diabetes (T2D) or coronary heart disease (CHD). We studied 2,411 participants of the Framingham Offspring Study (mean age 60 years, 52% women) for quantitative traits and CHD, and the Framingham SHARe database for T2D risk. We genotyped 19 single-nucleotide polymorphisms (SNPs) that tagged 85% (at r(2) = 0.8) of common (>5%) CNR1 SNPs. Fasting blood glucose and insulin at the 7th (1999-2001) exam were collected. We used age-, sex-, BMI-adjusted models to test additive associations of genotype with homeostasis model assessment of insulin resistance (HOMA(IR)) (linear mixed-effect models), T2D, or CHD. To account for multiple tests of SNPs, we generated empirical P values. The C allele at SNP rs806365 (frequency, 57.4%), ~4.1 kb 3' from CNR1, was associated with increased HOMA(IR) (n = 2,261, beta = 0.05 per C, empirical P = 0.01), risk of T2D (674 cases, odds ratio = 1.19 per C, nominal P = 0.01) and CHD (237 cases, hazard ratio = 1.23 per C, nominal P = 0.04). The association of rs806365 with HOMA(IR) was replicated in a meta-analysis of two independent cohorts (National Health and Nutrition Examination Survey III genetic cohort (NHANES-III) plus Partners Case Control Diabetes Study; 2,540 white individuals, beta = 0.037, nominal P = 0.007), but not in the large Meta-Analyses of Glucose and Insulin-related traits Consortium (MAGIC) Consortium (n = 29,248, nominal P = 0.74). The association of rs806365 was not replicated either with T2D in Diabetes Genetics Replication and Meta-analysis (DIAGRAM) (n = 10,128, nominal P = 0.31), or with CHD in PROCARDIS (n = 13,614, nominal P = 0.37). Although supported by initial results, we found no reproducible statistical association of common variation at CNR1 with insulin resistance, T2D, or CHD. PMID- 21633407 TI - Combination inhibition of the renin-angiotensin system: is more better? AB - Angiotensin-converting enzyme inhibitors or angiotensin II receptor blockers are considered the standard of care for treatment of cardiovascular disease and chronic kidney disease. Combination therapy with both angiotensin-converting enzyme inhibitors and angiotensin II receptor blockers effectively inhibits the renin-angiotensin system as well as potentiates the vasodilatory effects of bradykinin. It has been advocated that this dual blockade approach theoretically should result in improved clinical outcomes in both cardiovascular disease and chronic kidney disease. Clinical trial evidence for the use of combination therapy with angiotensin-converting enzyme inhibitors and angiotensin II receptor blockers in cardiovascular disease has provided conflicting results in hypertension, congestive heart failure, and ischemic heart disease. Clinical trial evidence to support combination therapy with angiotensin-converting enzyme inhibitors and angiotensin II receptor blockers in chronic kidney disease has largely been based on proteinuria reduction as a surrogate marker for clinically meaningful outcomes. Recent large-scale randomized clinical trials have not been able to validate protection in halting progression in chronic kidney disease with a dual blockade approach. This review serves as an appraisal on the clinical evidence of combination angiotensin-converting enzyme inhibition and angiotensin II receptor blockade in both cardiovascular disease and chronic kidney disease. PMID- 21633408 TI - Calcimimetics inhibit renal pathology in rodent nephronophthisis. AB - The development and progression of renal cysts appears to be driven by reduced cellular calcium and increased cyclic adenosine monophosphate (cAMP) from G protein-coupled receptors. To test whether treatment with a calcimimetic that stimulates the G-protein-coupled calcium-sensing receptor might normalize cystic epithelial cell intracellular calcium and cAMP, thereby inhibiting cyst progression, we used pcy mice. These animals develop cysts principally in the collecting duct, as do humans with nephronophthisis (NPHP). We administered the calcimimetic R-568 mixed in their food at early or late stages in the pathogenesis of cyst formation. The treatment reduced cyst enlargement, and the early treatment inhibited development of renal fibrosis. Although the effect of later treatment was more modest, both stages of the disease responded positively to treatment. Additionally, R-568 decreased total kidney cAMP in the pcy mice and, in vitro, decreased cAMP levels and cell proliferation, while increasing intracellular calcium in immortalized human autosomal recessive polycystic kidney disease renal epithelial cells. The latter two effects were unique to R-568 and not replicated by raising extracellular calcium. Thus, treating pcy mice with R 568 was effective in reducing cyst progression in this rodent model of NPHP. Direct studies will be needed to determine whether these results can be applied to the human disease. PMID- 21633410 TI - Low exposure to melamine increases the risk of urolithiasis in adults. AB - Melamine, a widely used chemical found in many products in daily use, became a public health concern due to melamine-associated urinary stone formation in children. In adults, it is still unknown whether low-dose melamine exposure may also cause urolithiasis. To address this question, we studied 211 Taiwanese patients diagnosed with calcium urolithiasis and 211 age- and gender-matched controls. All patients completed a detailed questionnaire and provided blood and urine samples for biochemical analysis. Urinary melamine concentrations were measured by triple-quadrupole liquid chromatography tandem mass spectrometry. Compared with those whose urinary melamine levels were below the detection limit of the method, patients with urinary melamine levels of up to 3.11 ng/ml and those with levels of >=3.12 ng/ml had 3.01- and 7.64-fold increased risk, respectively, of calcium urolithiasis after adjusting for educational level, fluid intake, cigarette smoking, betel quid chewing, alcohol drinking, urinary uric acid, calcium, creatinine, and estimated creatinine clearance rate. The population attributable risk of calcium urolithiasis averaged 50% when melamine was detected in the urine, after considering other covariates. MALDI-TOF mass spectrometry detected melamine in the stones of nine representative patients who had measurable urinary melamine levels. Thus, low-dose melamine exposure can play an important role in calcium urolithiasis in Taiwanese adults. PMID- 21633412 TI - Atmospheric observations of multiple molecular species using ultra-high resolution external cavity quantum cascade laser heterodyne radiometry. AB - We demonstrate a widely tunable laser heterodyne radiometer operating in the thermal IR during an atmospheric observation campaign in the solar occultation viewing mode. An external cavity quantum cascade laser tunable within a range of 1120 to 1238 cm(-1) is used as the local oscillator (LO) of the instrument. Ultra high-resolution (60 MHz or 0.002 cm(-1) transmission spectroscopy of several atmospheric species (water vapor, ozone, nitrous oxide, methane, and dichlorodifluoromethane) has been demonstrated within four precisely selected molecule-specific narrow spectral windows (~1 cm(-1). Atmospheric transmission lines within each selected window were fully resolved through mode-hop-free continuous tuning of the LO frequency. Comparison measurements were made simultaneously with a high-resolution Fourier transform spectrometer to demonstrate the advantages of the laser heterodyne system for atmospheric sounding at high spectral and spatial resolutions. PMID- 21633409 TI - Elevated depressive affect is associated with adverse cardiovascular outcomes among African Americans with chronic kidney disease. AB - This study was designed to examine the impact of elevated depressive affect on health outcomes among participants with hypertensive chronic kidney disease in the African-American Study of Kidney Disease and Hypertension (AASK) Cohort Study. Elevated depressive affect was defined by Beck Depression Inventory II (BDI-II) thresholds of 11 or more, above 14, and by 5-Unit increments in the score. Cox regression analyses were used to relate cardiovascular death/hospitalization, doubling of serum creatinine/end-stage renal disease, overall hospitalization, and all-cause death to depressive affect evaluated at baseline, the most recent annual visit (time-varying), or average from baseline to the most recent visit (cumulative). Among 628 participants at baseline, 42% had BDI-II scores of 11 or more and 26% had a score above 14. During a 5-year follow-up, the cumulative incidence of cardiovascular death/hospitalization was significantly greater for participants with baseline BDI-II scores of 11 or more compared with those with scores <11. The baseline, time-varying, and cumulative elevated depressive affect were each associated with a significant higher risk of cardiovascular death/hospitalization, especially with a time-varying BDI-II score over 14 (adjusted HR 1.63) but not with the other outcomes. Thus, elevated depressive affect is associated with unfavorable cardiovascular outcomes in African Americans with hypertensive chronic kidney disease. PMID- 21633413 TI - Dynamic sample imaging in coherent diffractive imaging. AB - As the resolution in coherent diffractive imaging improves, interexposure and intraexposure sample dynamics, such as motion, degrade the quality of the reconstructed image. Selecting data sets that include only exposures where tolerably little motion has occurred is an inefficient use of time and flux, especially when detector readout time is significant. We provide an experimental demonstration of an approach in which all images of a data set exhibiting sample motion are combined to improve the quality of a reconstruction. This approach is applicable to more general sample dynamics (including sample damage) that occur during measurement. PMID- 21633411 TI - Early interleukin 6 production by leukocytes during ischemic acute kidney injury is regulated by TLR4. AB - Although leukocytes infiltrate the kidney during ischemic acute kidney injury (AKI) and release interleukin 6 (IL6), their mechanism of activation is unknown. Here, we tested whether Toll-like receptor 4 (TLR4) on leukocytes mediated this activation by interacting with high-mobility group protein B1 (HMGB1) released by renal cells as a consequence of ischemic kidney injury. We constructed radiation induced bone marrow chimeras using C3H/HeJ and C57BL/10ScNJ strains of TLR4 (-/-) mice and their respective TLR4 (+/+) wild-type counterparts and studied them at 4 h after an ischemic insult. Leukocytes adopted from TLR4 (+/+) mice infiltrated the kidneys of TLR4 (-/-) mice, and TLR4 (-/-) leukocytes infiltrated the kidneys of TLR4 (+/+) mice but caused little functional renal impairment in each case. Maximal ischemic AKI required both radiosensitive leukocytes and radioresistant renal parenchymal and endothelial cells from TLR4 (+/+) mice. Only TLR4 (+/+) leukocytes produced IL6 in vivo and in response to HMGB1 in vitro. Thus, following infiltration of the injured kidney, leukocytes produce IL6 when their TLR4 receptors interact with HMGB1 released by injured renal cells. This underscores the importance of TLR4 in the pathogenesis of ischemic AKI. PMID- 21633414 TI - High-resolution, background-free, time-to-space conversion by collinearly phase matched sum-frequency generation. AB - We report the first demonstration, to our knowledge, of time-to-space conversion of 1.55 MUm femtosecond optical pulses using nondegenerate, collinearly phase matched sum-frequency generation. A quasi-monochromatic and background-free output signal spanning a time window of 35 ps and with a pulse image width of 350 fs was achieved. The resulting serial-to-parallel resolution factor of 100 demonstrates the potential for all-optical complete frame demultiplexing of a 1 Tbit/s optical time-division multiplexing bit stream. PMID- 21633415 TI - Off-axis metallic diffractive lens for terahertz beams. AB - A diffractive optical element for off-axis focusing of terahertz radiation is presented. It was designed in a nonparaxial regime and manufactured in a metal slab by laser cutting of curved stripes. The optical function of the structure includes focusing and deflecting the illuminating beam of a chosen frequency in a particular place. Therefore, the element acts as both a spatial and a spectral filter; hence it is especially suitable for separating the terahertz signal from a broadband thermal load in passive detection devices. The experimental evaluation of the proposed diffractive lens by means of time-domain spectroscopy is presented and discussed. PMID- 21633416 TI - Two-dimensional scanning probe driven by a solenoid-based single actuator for optical coherence tomography. AB - We propose the two-dimensional scanning probe operating with a single actuator, which is thought useful as a sample probe for optical coherence tomography (OCT). The probe was designed to use a single-body lensed fiber cantilever loaded with an iron-bead and driven by a single-solenoid actuator. Elliptic spiral trace patterns were achieved using off-axis magnetic fields of the solenoid. A three dimensional OCT image was obtained for a scanning area of 3.8 mm * 3.4 mm at an acquisition speed of 16.7 s/V. Up to 27 Hz B-scan rate, the proposed probe worked well, and 1000 A-scans were made per each B-scan. PMID- 21633417 TI - Multiwatt level output powers from a tunable fiber optical parametric oscillator. AB - We present an all-fiber high average power fiber optical parametric oscillator based on standard telecommunications dispersion-shifted fiber. The output of the oscillator is continuously tunable out to +/-28 THz from the pump wavelength. The average power of the oscillator's output is in excess of 1.9 W in each sideband out to +/-25 THz detuning. Between 5 and 14 THz detuning, the average power of the Stokes output is in excess of 3.8 W. PMID- 21633418 TI - Surface-plasmon-polariton-assisted dipole-dipole interaction near metal surfaces. AB - We investigate the surface plasmon polariton (SPP)-assisted interaction between two dipoles near a metal surface. The radiation energy from a dipole can excite SPPs and transport to another dipole through the channel of the localized SPP modes. This energy transfer can be much more efficient than direct energy transfer via dipole-dipole radiation interaction in free space. A simple analytical model is proposed to describe the underlying physics behind the influence of SPP on the dipole-dipole interaction energy, and it predicts a wide variety of complicated interaction features that agree well with rigorous calculations. PMID- 21633419 TI - Common-path achromatic rotational-shearing coronagraph. AB - To suppress starlight for direct exoplanet observation, we propose a common-path achromatic rotational-shearing coronagraph (CP-ARC), which is an interferocoronagraph with an angular-adjustable field rotator. The CP-ARC aims to maintain unwanted detection of stellar light, which can be suppressed incompletely by interference because of the finite diameter of the star. Compared to the previous interferocoronagraph, which had a nonadjustable 180 degrees field rotation, the proposed CP-ARC can improve the coronagraphic contrast by several orders if the CP-ARC is combined with medium or large telescopes where the companion-star separation is optically resolved by more than a few Airy radii. The CP-ARC is made robust against mechanical disturbances due to the common-path interferometer principle. PMID- 21633420 TI - Porphyrin-doped solgel-lined structured optical fibers for local and remote sensing. AB - We constructed a type of sensor by depositing a solgel layer within the interior holes of a silica-structured fiber and, subsequently, coating this with an acid responsive porphyrin. Protonation of the porphyrin by an acidic gas (HCl in this case), is detected by a large change in the visible spectrum. Compared to previous work on a liquid-core sensor in a structured optical fiber, the signal to-noise ratio of this gas sensor shows a reduced signal strength, but the detection rate is increased about fortyfold. PMID- 21633421 TI - 160 Gbit/s binary-to-quaternary amplitude shift keying encoding in the optical domain. AB - 160 Gbit/s all-optical binary-to-quaternary amplitude shift keying format conversion is carried out in a nonlinear optical fiber. This scheme, which also acts as a 2 bit digital-to-analog convertor, has been confirmed through Q-factor measurements. PMID- 21633422 TI - Real-time holography in ruthenium-doped bismuth sillenite crystals at 1064 nm. AB - Real-time holographic recording and an improvement of the response time in ruthenium (Ru)-doped Bi(12)SiO(20) (BSO) crystal at 1064 nm is obtained. Using green light pre-exposure, a significant operation speed of 60 ms is achieved. In addition, the ability for image reconstruction is demonstrated in Ru-doped BSO, supporting further applications as reversible media for real-time image processing at the near-IR spectral range. PMID- 21633423 TI - All-optical switching at the Fano resonances in subwavelength gratings with very narrow slits. AB - We theoretically discuss all-optical switching at the Fano resonances of subwavelength gratings made of a chalcogenide glass (As(2)S(3)). Particular attention is devoted to the case in which the grating possesses extremely narrow slits (channels ranging from a~10 nm to a~40 nm). The remarkable local field enhancement available in these situations conspires to yield low-threshold switching intensities (~50 MW/cm(2)) at telecommunication wavelengths for extremely thin (d~200 nm) gratings when a realistic value of the As(2)S(3) cubic nonlinearity is used. PMID- 21633424 TI - 1 kW peak power passively Q-switched Nd(3+)-doped glass integrated waveguide laser. AB - Embedded optical sensors always require more compact, stable, and powerful laser sources. In this Letter, we present a fully integrated passively Q-switched laser, which has been realized by a Ag(+)/Na(+) ion exchange on a Nd(3+)-doped phosphate glass. A BDN-doped cellulose acetate thick film is deposited on the waveguide, acting as an upper cladding and providing a distributed saturable absorption. At lambda=1054 nm, the device emits pulses of 1.3 ns FWHM with a repetition rate of 28 kHz. These performances, coupled with the 1 kW peak power, are promising for applications such as supercontinuum generation. PMID- 21633425 TI - Simultaneous dual-wavelength-band common-path swept-source optical coherence tomography with single polygon mirror scanner. AB - We report a novel (to the best of our knowledge) simultaneous 1310/1550 two wavelength band swept laser source and dual-band common-path swept-source optical coherence tomography (SS-OCT). Synchronized dual-wavelength tuning is performed by using two laser cavities and narrowband wavelength filters with a single dual window polygonal scanner. Measured average output powers of 60 and 27 mW have been achieved for the 1310 and 1550 nm bands, respectively, while the two wavelengths were swept simultaneously from 1227 to 1387 nm for the 1310 nm band and from 1519 to 1581 nm for the 1550 nm band at an A-scan rate of 65 kHz. Broadband wavelength-division multiplexing is used for coupling two wavelengths into a common-path single-mode GRIN-lensed fiber probe to form dual-band common path SS-OCT. Simultaneous OCT imaging at 1310 and 1550 nm is achieved. This technique allows for in vivo high-speed OCT imaging with potential application in functional (spectroscopic) investigations. PMID- 21633426 TI - Continuous-wave terahertz digital holography by use of a pyroelectric array camera. AB - Terahertz (THz) digital holography is realized based on a 2.52 THz far-IR gas laser and a commercial 124 * 124 pyroelectric array camera. Off-axis THz holograms are obtained by recording interference patterns between light passing through the sample and the reference wave. A numerical reconstruction process is performed to obtain the field distribution at the object surface. Different targets were imaged to test the system's imaging capability. Compared with THz focal plane images, the image quality of the reconstructed images are improved a lot. The results show that the system's imaging resolution can reach at least 0.4 mm. The system also has the potential for real-time imaging application. This study confirms that digital holography is a promising technique for real-time, high-resolution THz imaging, which has extensive application prospects. PMID- 21633427 TI - Observation of microsphere movement driven by optical pulse. AB - A laser beam was coupled into a tapered optical fiber tip with a diameter of about 2.6 MUm, and a 46 MUm diameter microsphere was propelled by the outgoing optical pulse. With the change of pulse energy from 1.35 to 7.22 MUJ, the calculated average velocity of the driven microsphere varied from 0.38 to 10.68 cm/s. The scanning electron microscope images of the fiber tip show that there is no thermal damage during the experiment. PMID- 21633428 TI - Collective modulation instability of multiple four-wave mixing. AB - We investigate the modulation instability of multiple four-wave mixing arising from a dual-frequency pump in a single-mode fiber or waveguide. By applying the Floquet theory on account of the periodic nature of four-wave mixing, we reveal a collective type of instability occurring in the anomalous dispersion regime. Our interpretation of the linear stability analysis is validated by the numerical solution of the nonlinear Schrodinger equation. PMID- 21633429 TI - Asymmetric resonance in selective reflection: explanation via Fano-like mechanism. AB - Fano mechanism is the universal explanation of asymmetric resonance appearing in different systems. We report the evidence of Fano-like resonance in selective reflection from a resonant two-level medium. We draw an analogy with the asymmetric resonance previously obtained for the coupled oscillators. We also take into account the effects of dielectric background and local-field correction and connect our results with optical bistability. PMID- 21633430 TI - An absolute test for axicon surfaces. AB - We present a method for absolute testing of axicon surfaces in a null test setup. The absolute test exploits the symmetry of axicons, which allows us to introduce a shift of the surface under test in both the axial and rotational directions while still maintaining the null test condition. With two shifts of the surface under test and four measurements, the interferometer and null optics error can be removed. The absolute surface local deviation can be obtained by wavefront reconstruction with a double-side spiral-path direct integration method. A simulation of the method, including typical systematic as well as statistical errors as input, is presented to estimate the error propagation. Experimental absolute test results of a 90 degrees axicon surface are given. PMID- 21633431 TI - Refractive index measurement of optical waveguides using modified end-fire coupling method. AB - We present a modified end-fire coupling method to reconstruct two-dimensional index profiles of optical waveguides. The reconstruction is based on the measured differential optical fields and an inverse algorithm for the Helmholtz equation. The differential fields are obtained from spatially perturbed images recorded by a CCD camera. A closed-loop piezoelectric stage given a sinusoidal wave and an oil-immersion objective lens for high-resolution images are used in the measurement. A single-mode fiber and a Ti:LiNbO(3) waveguide with known index profile at 632.8 nm are measured for demonstration. The measured index profiles agree quite well with core regions of known index profiles. PMID- 21633432 TI - Ultrasmall subwavelength nanorod plasmonic cavity. AB - We propose an ultrasmall plasmonic cavity consisting of a high-index/low-index dielectric nanorod covered with silver. Full three-dimensional subwavelength confinement of the surface-plasmon polaritons was achieved at the high-index dielectric-silver interface without propagating to the low-index dielectric silver interface. The numerical simulations showed that the plasmonic mode excited in this cavity has a deep subwavelength mode volume of 0.0038(lambda/2n)(3) and a quality factor of 1500 at 40 K, and consequently a large Purcell factor of ~2*10(5). Therefore, this plasmonic cavity is expected to be useful for the demonstration of high-efficiency single photon sources or low threshold lasers in an ultracompact nanophotonic circuit. PMID- 21633433 TI - Tight focusing of a double-ring-shaped, azimuthally polarized beam. AB - We study the focusing properties of a double-ring-shaped azimuthally polarized beam through an annular high NA objective lens. It is shown that a subwavelength focal hole (~0.5lambda) with a quite long depth of focus (~26lambda) is achieved near the focus. This kind of nondiffracting focal hole is called dark channel, which may have applications in atom optical experiments, such as with atomic lenses, atom traps, and atom switches. PMID- 21633434 TI - Measuring the topological charge of optical vortices with an axicon. AB - We analyzed the spatial spectrum of the diffraction intensity pattern of an ideal Bessel beam and found an implicit rule that the number of the bright rings in the spatial spectrum is equal to the topological charge of the Bessel beam. The radius of the bright and dark rings has some relation with the topological charge and can be determined accurately. It provides us with a new way for measuring the topological charge of an optical vortex through its diffraction intensity pattern after an axicon. The results of simulation coincide with the theory. PMID- 21633435 TI - Doppler velocimetry on microparticles trapped and propelled by laser light in liquid-filled photonic crystal fiber. AB - Laser Doppler velocimetry is used to measure very accurately the velocity and position of a microparticle propelled and guided by laser light in liquid-filled photonic crystal fiber. Periodic variations in particle velocity are observed that correlate closely with modal beating between the two lowest order guided fiber modes. PMID- 21633436 TI - Proposal and demonstration of a spectrometer using a diffractive optical element with dual dispersion and focusing functionality. AB - We propose an optical spectrometer using a hybrid grating-Fresnel (G-Fresnel) diffractive optical element. Theoretical simulation shows that a spectral resolution of approximately 1 nm can be potentially achieved with a millimeter sized G-Fresnel. A proof-of-concept G-Fresnel-based spectrometer with subnanometer spectral resolution is experimentally demonstrated. The proposed method provides a promising new way for realizing compact optical spectrometers. PMID- 21633437 TI - Widely tunable extreme UV frequency comb generation. AB - Extreme UV (XUV) frequency comb generation in the wavelength range of 51 to 85 nm is reported based on high-order harmonic generation of two consecutive IR frequency comb pulses that were amplified in an optical parametric chirped pulse amplifier. The versatility of the system is demonstrated by recording direct XUV frequency comb excitation signals in He, Ne, and Ar with visibilities of up to 61%. PMID- 21633438 TI - Plasmonically enhanced optical transmission through a metalized nanostructured photonic crystal fiber taper. AB - A subwavelength holey plasmonic structure was proposed by adiabatically tapering a photonic crystal fiber (PCF) and subsequently metalizing the cleaved end facet. By coupling a white light into the PCF side, we experimentally observed an enhanced optical transmission in the spectral domain through the plasmonic structure at the tapered end. We further showed numerically that the proposed device renders a focused directional beam, due to its Fresnel-zone-like configuration and the plasmonic lensing effects. PMID- 21633439 TI - Variable aperture stop based on the design of a single chamber silicone membrane lens with integrated actuation. AB - In this Letter we report on the fabrication and testing of an extremely thin variable aperture stop based on the design of a single chamber adaptive membrane lens with integrated actuation. The aperture consists of a ring-shaped piezoelectric bending actuator with an elastic silicone membrane in the center. The formed cavity is filled with a nontransparent fluid and sealed with a glass platelet. In a voltage range up to 80 V, an opening of the aperture of 4.55 mm in diameter is obtained. The transmission in comparison to a standard mechanical aperture stop is maximum 6% lower. PMID- 21633440 TI - Calculating depth maps from digital holograms using stereo disparity. AB - Depth extraction is an important aspect of three-dimensional (3D) image processing with digital holograms and an essential step in extended focus imaging and metrology. All available depth extraction techniques with macroscopic objects are based on variance; however, the effectiveness of this is object dependent. We propose to use disparity between corresponding points in intensity reconstructions to determine depth. Our method requires a single hologram of a scene, from which we reconstruct two different perspectives. In the reconstruction the phase information is not needed, which makes this method useful for in-line digital holography. To our knowledge disparity based 3D image processing has never been proposed before for digital holography. PMID- 21633441 TI - High sensitive determination of laser-induced frequency shifts of ultracold cesium molecules. AB - We experimentally present a technique for sensitively determining the laser induced frequency shifts of the long-range molecular vibrational and rotational levels. The scheme relies on an optical frequency shifter, leading to two laser beams with a precise and adjustable frequency interval. A series of photoassociation spectra are recorded with both beams inducing molecular lines, whose peak separation provides an accurate frequency ruler to measure the frequency shifts of cesium molecular levels, which have not yet been observed in previous reports. The data are compared to theoretical predictions and show a good agreement. PMID- 21633442 TI - Spectral phase retrieval of 8 fs optical pulses at 600 nm by using a collinear autocorrelator with 300-MUm-thick lithium triborate crystals. AB - We report on noniterative spectral phase retrieval of 1.1 nJ, 8 fs pulses at 600 nm by using 300-MUm-thick lithium triborate crystals in a standard collinear autocorrelator with ~2 min data acquisition time. This method is simple, sensitive, and immune to the spectral distortion and UV absorption of the linear and nonlinear optics. PMID- 21633443 TI - Highly coherent terahertz wave generation with a dual-frequency Brillouin fiber laser and a 1.55 MUm photomixer. AB - Thanks to a portable dual-frequency Brillouin fiber laser and a 1.55 MUm photomixer, we report the generation of a highly coherent kilohertz level submillimeter wave emission. Low-cost telecommunications components are used to achieve very simple source architecture. The photomixer is composed of a unitravelling carrier photodiode integrated with an antenna. An emission at 316 GHz is observed and analyzed thanks to heterodyne detection with a signal-to noise ratio >65 dB and a ~1 kHz linewidth. The phase noise of the proposed source has the same performance at 1.7 and 316 GHz. We show that this source has comparable or better phase noise compared to electrical oscillators and the tunability is much wider. PMID- 21633444 TI - Split-pulse spectrometer for absolute XUV frequency measurements. AB - A split-pulse spectrometer based on pairs of time-delayed femtosecond pulses can give access to accurate frequency measurements in the extreme ultraviolet (XUV) spectral domain. We demonstrate this approach by measuring the absolute frequency of a single-XUV-photon transition to a bound state of atomic argon excited with the ninth harmonic of an amplified Ti:sapphire laser. PMID- 21633445 TI - Tunable synchronously-pumped fiber Raman laser in the visible and near-infrared exploiting MOPA-generated rectangular pump pulses. AB - We report a tunable synchronously pumped fiber Raman laser (SPFRL) in the near infrared (NIR) and visible wavebands pumped by a pulsed, all-fiber PM 1060 nm master oscillator power amplifier (MOPA) and its frequency-doubled output, respectively. The seed was adaptively shaped to deliver rectangular output pulses, thereby enabling selective excitation of individual Raman Stokes lines. Using filtered synchronous feedback of the desired Raman Stokes line, the linewidth of the SPFRL was reduced by a factor of 4 and the extinction ratio of the desired Raman Stokes was improved by more than 3 dB relative to a simple single-pass conversion scheme. A continuous tuning range of 2.2 THz was obtained for each of the Raman Stokes orders in the visible (spanning from green to orange first to fifth Stokes lines). A larger 5.0 THz tunable range was achieved in the NIR spectral region. PMID- 21633446 TI - Photoacoustic-guided convergence of light through optically diffusive media. AB - We demonstrate that laser beams can be converged toward a light-absorbing target through optically diffusive media by using photoacoustic-guided interferometric focusing. The convergence of light is achieved by shaping the wavefront of the incident light with a deformable mirror to maximize the photoacoustic signal, which is proportional to the scattered light intensity at the light absorber. PMID- 21633447 TI - In-fiber integrated accelerometer. AB - A compact in-fiber integrated fiber-optic Michelson interferometer based accelerometer is proposed and investigated. In the system, the sensing element consists of a twin-core fiber acting as a bending simple supported beam. By demodulating the optical phase shift, we obtain that the acceleration is proportional to the force applied on the central position of the twin-core fiber. A simple model has been established to calculate the sensitivity and resonant frequency. The experimental results show that such an accelerometer has a sensitivity of 0.09 rad/g at the resonant frequency of 680 Hz. PMID- 21633448 TI - Three-dimensional polarization-independent visible-frequency carpet invisibility cloak. AB - We miniaturize all features in a previously introduced polarization-independent three-dimensional carpet invisibility cloak by more than a factor of 2. This leads to operation wavelengths in the visible. The structures are characterized by electron and optical microscopy. In contrast to our previous work at IR wavelengths, we can directly measure two-dimensional images at visible frequencies, perform control experiments from the backside, and compare the images with theory. We find excellent agreement. Furthermore, we study the wavelength dependence in the range from 900 nm down to 500 nm. Cloaking action deteriorates as the woodpile stop band at around 575 nm is approached. PMID- 21633449 TI - High-repetition-rate distributed Brillouin sensor based on optical correlation domain analysis with differential frequency modulation. AB - A kind of high-repetition-rate distributed Brillouin sensor is proposed and experimentally demonstrated based on optical correlation-domain analysis with differential frequency modulation, where the optical frequencies of the pump and the probe waves are modulated at slightly different RFs so that the temporal position of the measurement is continuously and repeatedly swept along a fiber under test. A distribution map of Brillouin frequency variation along a 100 m optical fiber is acquired at a repetition rate of 20 Hz with an accuracy of +/ 2.5 MHz and a spatial resolution of about 80 cm. PMID- 21633450 TI - Observation of asymptotic localization of a wave packet. AB - We present the experimental observation of asymptotic localization, a new regime of wave packet localization under the action of an external linear potential gradient. We employ arrays of evanescently coupled waveguides and directly observe the wave packet trajectory, which exhibits an oscillation with vanishing amplitude and period. PMID- 21633451 TI - Principal-component-analysis-based estimation of blood flow velocities using optical coherence tomography intensity signals. AB - The intensity signal in optical coherence tomography contains information about the translational velocity of scatterers, and can be used to quantify blood flow. We apply principal component analysis to efficiently extract this information. We also study use of nonuniform temporal sampling of the intensity signal to increase the range of quantifiable flow velocities. We demonstrate this technique in simulation, phantom and in vivo blood flow measurements, and highlight its potential to enable three-dimensional wide-field mapping of blood flow using OCT. PMID- 21633452 TI - Preconditioning for multidimensional TOMBO imaging. AB - In this Letter, we propose a preconditioning method to improve the convergence speed of iterative reconstruction algorithms in a compact, multidimensional, compound-eye imaging system called the thin observation module by bound optics. The condition number of the system matrix is improved by using a preconditioner matrix. To calculate the preconditioner matrix, the system model is expressed in the frequency domain. The proposed method is simulated by using a compressive sensing algorithm called the two-step iterative shrinkage/thresholding algorithm. The results showed improved reconstruction fidelity with a certain number of iterations for high signal-to-noise ratio measurements. PMID- 21633453 TI - Broadband micro-Michelson interferometer with multi-optical-path beating using a sphered-end hollow fiber. AB - We demonstrate a high-sensitivity broadband (1250-1650 nm) fiber micro-Michelson interferometer using a single-mode fiber end-spliced with a sphered-end hollow core fiber. The hollow core is slightly smaller than the solid core of a single mode fiber, so the fractional power of the core mode is converted into cladding modes. The excited cladding modes propagate at distinct optical paths along the hollow-core fiber and have individual foci outside the spherical lens. The reflected core mode, generated at the solid core-air interface, and the reflected cladding modes, generated at external material, interfere with each other to produce beating in the interference signals. PMID- 21633454 TI - Dual-mode operation of a twisted nematic liquid crystal cell by switching between dynamic and memory modes. AB - We propose a twisted nematic liquid crystal device that can be operated in dynamic or memory mode, based on the information content to be displayed at that time. +90 degrees -twisted and -90 degrees -twisted states are used as two stable states for operation in the memory mode. A vertical electric field is applied to realize gray levels for operation in the dynamic mode. The proposed device has a memory retention time of over a month for the memory mode and a response time of 12 ms for the dynamic mode. Contrast ratios of over 500?1 can be obtained in both the dynamic and memory modes. PMID- 21633455 TI - Continuous-wave fiber cavity ring-down measurements using frequency-shifted interferometry. AB - We present a spatial-domain fiber cavity ring-down (CRD) technique that does not require optical pulses for time-resolved detection. Instead of measuring the intensity decay rate of an optical pulse in the time domain as in conventional CRD experiments, we measure the decay rate of a CW signal in a ring-down cavity (RDC) using frequency-shifted interferometry in the spatial domain, as a function of distance traveled by the light. As a proof-of-concept demonstration, we measured fiber bend loss in a loop RDC using this technique, and a cavity loss change as low as 0.0135 dB induced by every fiber turn on a mandrel was unambiguously discerned. PMID- 21633456 TI - Markov chain formalism for vector radiative transfer in a plane-parallel atmosphere overlying a polarizing surface. AB - We report on a way of building bidirectional surface reflectivity into the Markov chain formalism for polarized radiative transfer through a vertically inhomogeneous atmosphere. Numerical results are compared to those obtained by the Monte Carlo method, showing the accuracy of the Markov chain method when 90 streams are used to compute the radiation from a Rayleigh-plus-aerosol atmosphere that overlies a surface with a bidirectional reflection function consisting of both depolarizing and polarizing parts. PMID- 21633457 TI - Discriminating thermal effect in nonlinear-ellipse-rotation-modified Z-scan measurements. AB - We report that a modified Z-scan method by nonlinear ellipse rotation (NER) can be used to discriminate true nonlinear refraction from thermal effect in the transient regime and steady state. The combination of Z-scan and NER allows us to measure the third-order nonlinear susceptibility component without the influence of thermal-optical nonlinearity. The experimental results of pure CS(2) and CS(2) solutions of nigrosine verify that the transient thermal effect can be successfully eliminated from the NER-modified Z-scan measurements. This method is also extended to the case in which thermal-optical nonlinearities depend on a high repetition rate of femtosecond laser pulses for the N,N-dimethylmethanamide solutions of graphene oxide. PMID- 21633458 TI - Bidirectional passively mode-locked soliton fiber laser with a four-port circulator. AB - We present an all-fiber bidirectional passively mode-locked soliton laser with what we believe is a novel cavity configuration. Using a four-port circulator, we incorporate two different semiconductor saturable absorber mirrors (SESAMs) into the laser cavity, which enables bidirectional mode locking. The laser allows the generation of two independent countercirculating mode-locked pulse trains, each with an individual fundamental repetition rate that can be adjusted by varying the SESAM pigtail length. Two countercirculating pulse trains with repetition rates of 21.3 and 15.2 MHz are obtained simultaneously. By controlling the intracavity loss imposed on these two pulse trains, either one of the two pulse trains can be switched on or off. The bidirectional operation with other repetition rates is also demonstrated. PMID- 21633459 TI - Significant retrieval of lost evanescent power by tuning modes close-to-cutoff with a gel-coated taper. AB - We report a fiber-optic evanescent-wave (EW) sensor capable of dramatically increasing the power collection level by capturing the EW power that is normally lost. The key element is a taper with a thin overlay that is completely separated from the sample by an arbitrary distance and thus operates remotely within the instrumentation system. A two-stage tuning of the close-to-cutoff modes occurring within this element is proposed to interpret the observed phenomenon. PMID- 21633460 TI - Fast perturbation Monte Carlo method for photon migration in heterogeneous turbid media. AB - We present a two-step Monte Carlo (MC) method that is used to solve the radiative transfer equation in heterogeneous turbid media. The method exploits the one-to one correspondence between the seed value of a random number generator and the sequence of random numbers. In the first step, a full MC simulation is run for the initial distribution of the optical properties and the "good" seeds (the ones leading to detected photons) are stored in an array. In the second step, we run a new MC simulation with only the good seeds stored in the first step, i.e., we propagate only detected photons. The effect of a change in the optical properties is calculated in a short time by using two scaling relationships. By this method we can increase the speed of a simulation up to a factor of 1300 in typical situations found in near-IR tissue spectroscopy and diffuse optical tomography, with a minimal requirement for hard disk space. Potential applications of this method for imaging of turbid media and the inverse problem are discussed. PMID- 21633461 TI - Design of a wide-angle, lightweight head-mounted display using free-form optics tiling. AB - We present a concept of a wide-angle, lightweight, optical see-through head mounted display (HMD) using free-form optics tiling. Free-form optics tiling can potentially address several critical problems in existing tiled HMD designs that use rotationally symmetric optics. The optical design of our tiled optical see through HMD achieves a field of view (FOV) of 56 degrees * 45 degrees and an angular resolution of 3.2 arcmin with two display channels. We demonstrate a proof-of-concept prototype and present some of its manufacturing details. The FOV can be further enlarged by tiling more display channels together at their bottom and side surfaces. PMID- 21633462 TI - Efficient broadband silicon-on-insulator grating coupler with low backreflection. AB - We design and fabricate an efficient broadband grating coupler on a 400 nm thick silicon-on-insulator wafer. The measured coupling loss is 3 dB when coupling to a single-mode fiber at 1310 nm wavelength with TE polarization. The spectral FWHM and backreflection are determined to be 58 nm and -27 dB, respectively. PMID- 21633463 TI - Linear and nonlinear optical properties of carbon nanotube-coated single-mode optical fiber gratings. AB - Single-wall carbon nanotube deposition on the cladding of optical fibers has been carried out to fabricate an all-fiber nonlinear device. Two different nanotube deposition techniques were studied. The first consisted of repeatedly immersing the optical fiber into a nanotube supension, increasing the thickness of the coating in each step. The second deposition involved wrapping a thin film of nanotubes around the optical fiber. For both cases, interaction of transmitted light through the fiber core with the external coating was assisted by the cladding mode resonances of a tilted fiber Bragg grating. Ultrafast nonlinear effects of the nanotube-coated fiber were measured by means of a pump-probe pulses experiment. PMID- 21633464 TI - Artifact analysis and image enhancement in three-dimensional computational integral imaging using smooth windowing technique. AB - We propose an artifact analysis in computational integral imaging and the image enhancement method based on the analysis using the smooth windowing technique. Blurring and lenslet artifacts, which are major problems in computational integral imaging, are defined and analyzed using a signal model. Applying a smooth and continuous window such as the triangular window to computational integral imaging reconstruction provides a dramatic improvement in terms of image quality. Experimental results are presented to show the validity of our method. To our best knowledge, this is the first trial to control a window function in computational integral imaging. PMID- 21633465 TI - Temperature-independent silicon subwavelength grating waveguides. AB - We demonstrate, by experiment and numerical calculations, temperature-independent subwavelength grating waveguides with a periodic composite core composed of alternating regions of silicon and SU-8 polymer. The polymer has a negative thermo-optic (TO) material coefficient that cancels the large positive TO effect of the silicon. Measurements and Bloch mode calculations were carried out over a range of silicon-polymer duty ratios. The lowest measured TO coefficient at a wavelength of 1550 nm is 1.8*10(-6) K(-1); 2 orders of magnitude smaller than a conventional silicon photonic wire waveguide. Calculations predict the possibility of complete cancellation of the silicon waveguide temperature dependence. PMID- 21633466 TI - Single-input spherical microbubble resonator. AB - A single-input whispering gallery optical microbubble resonator is presented. Spherical microbubbles with diameters less than 100 MUm, micrometer-sized wall thicknesses, and a single opening or input were fabricated by heating the tapered tip of a pressurized glass capillary using a CO(2) laser. Optical whispering gallery modes with Q factors of ~10(5) were obtained. The bubbles were filled with water and mode shifts of ~20 GHz were observed. Fano-type resonances were detected when the coupling optical fiber diameter was less than 1 MUm, causing the microresonator to switch from being a band-stop filter to a bandpass filter. Larger bubbles with submicrometer wall thickness were also fabricated. PMID- 21633467 TI - Differential laser-induced perturbation spectroscopy using a deep-ultraviolet excimer laser. AB - Here we demonstrate a unique optical sensing scheme based on deep-UV photochemical perturbation in combination with difference spectroscopy. Applying a sequence of optical probing, UV-laser-induced perturbation, and repeat optical probing coupled with difference spectroscopy provides a new spectral signature. We show a selective (sevenfold difference) optical response using a fluorescence probe for binary mixtures of organic dyes, and generate complementary spectral information derived from Raman scattering of the dipeptide glycine-glycine. We further extend the methodology to fluorescence-based imaging of an organic matrix. PMID- 21633468 TI - Real-time line projection for fast terahertz spectral computed tomography. AB - We demonstrated fast terahertz spectral computed tomography by using real-time line projection of a terahertz beam. Two types of cross-sectional images of continuously rotating samples have been measured in only a few seconds. From temporal data, a peak-to-peak sinogram and cross sections have been reconstructed using a filtered backprojection algorithm. Using fast Fourier transform from temporal data, spectral cross sections of the sample have been obtained. PMID- 21633469 TI - Mode-locked fiber laser using an SU8/SWCNT saturable absorber. AB - We report the fabrication of a saturable absorber based on SU8 single wall carbon nanotube (SWCNT) composite material. Thin films with a controllable thickness can be fabricated using a simple and reliable process. These films can be inserted between two FC/APC connectors in order to have an inline saturable absorber. A passive mode-locked laser was built by interleaving the fiberized saturable absorber in an erbium-doped fiber (L-band) ring cavity laser. The laser produces 871 fs pulses with a repetition rate of 21.27 MHz and a maximum average power of 1 mW. PMID- 21633470 TI - Femtosecond laser-induced inverted microstructures inside glasses by tuning refractive index of objective's immersion liquid. AB - We report the formation of inverted microstructures inside glasses after femtosecond laser irradiation by tuning the refractive index contrast between the immersion liquid and the glass sample. By using water as well as 1 bromonaphthalene as immersion liquids, microstructures with similar shape but opposite directions are induced after femtosecond laser irradiation. Interestingly, the elemental distribution in the induced structures is also inverted. The simulation of laser intensity distribution along the laser propagation direction indicates that the interfacial spherical aberration effect is responsible for the inversion of microstructures and elemental distribution. PMID- 21633471 TI - Fraunhofer computer-generated hologram for diffused 3D scene in Fresnel region. AB - A Fraunhofer computer-generated hologram (CGH) is proved to be valid in display for three-dimensional (3D) objects from the Fresnel to the far-field region without a Fourier lens for reconstruction. To quickly compute large and complicated 3D objects that consist of slanted diffused surfaces in the Fresnel region, a Fraunhofer-based analytical approach using a basic-triangle tiling diffuser is developed. Both theoretical and experimental results reveal that Fraunhofer CGH can perform the same effects as Fresnel CGH but require less calculation time. Impressive 3D solid effects are achieved in the Fresnel region. PMID- 21633472 TI - Nonreciprocal switching thresholds in coupled nonlinear microcavities. AB - A concept for the design of nonlinear optical diodes is proposed that uses the multistability of coupled nonlinear microcavities and the dependence of switching thresholds on the direction of incidence. A typical example of such a diode can be created by combining two mirror-symmetric microcavities where modes of the opposite parity dominate. It is shown that a strong nonreciprocal behavior can be achieved together with a negligible insertion loss. To describe the dynamical properties of such systems, a model based on the coupled-mode theory is developed, and a possible implementation in the form of multilayered structures is considered. PMID- 21633473 TI - Observation of polarization conflict caused by geometrical phase in a twisted nematic liquid crystal cell. AB - We analyze the optical effects associated with an adiabatic rotation of a plane of polarization in a twisted nematic liquid crystal. The experimental verification was performed with a cell with linear rubbing of a front surface and circular rubbing of a rear surface. The expectations of the liquid crystal's orientation defect origin along the line of the maximum tension and a polarization conflict caused by geometrical phase are confirmed. PMID- 21633474 TI - Elementary heterogeneously polarized field modeling. AB - An approach for experimental modeling of elementary field cells with heterogeneous polarization is proposed. It is shown that such cells may be obtained by the superposition of orthogonally linearly polarized waves with relatively simple phase surfaces and close intensities. The experimental arrangement for obtaining elementary field cells is suggested. The characteristics of a polarization cell may be easily changed by the change of superposed beam parameters. The results of experimental investigation are presented. PMID- 21633475 TI - Spectral dynamics of modulation instability described using Akhmediev breather theory. AB - The Akhmediev breather formalism of modulation instability is extended to describe the spectral dynamics of induced multiple sideband generation from a modulated continuous wave field. Exact theoretical results describing the frequency domain evolution are compared with experiments performed using single mode fiber around 1550 nm. The spectral theory is shown to reproduce the depletion dynamics of an injected modulated continuous wave pump and to describe the Fermi-Pasta-Ulam recurrence and recovery towards the initial state. Realistic simulations including higher-order dispersion, loss, and Raman scattering are used to identify that the primary physical factors that preclude perfect recurrence are related to imperfect initial conditions. PMID- 21633476 TI - Measurement of particle concentration in flow by statistical analyses of optical coherence tomography signals. AB - We present a statistic method to measure the concentration of particles in flow by the use of optical coherence tomography (OCT). This method is based on the fact that the fluctuating numbers of particles contained within a small OCT detection volume caused by Brownian motion are governed by Poissonian distribution. Particle concentration is evaluated from the statistical analyzes of the OCT intensity signals that are backscattered by the moving particles. The method is experimentally verified with flow phantoms of polymer microspheres. Experimental results are in good agreement with the theoretical ones for low concentrations while relatively large discrepancies emerge for high concentrations. PMID- 21633477 TI - Directional far-field response of a spherical nanoantenna. AB - We study the directional far-field response of a spherical nanoantenna via engineering the plasmonic nanosphere's distance, size, and material. A unified pattern synthesis approach based on the T-matrix method and the particle swarm optimization is proposed for the directional beamforming of the nanoantenna. The angular response of the directional nanoantenna is very sensitive to the material change but is immunized to the random error of the spatial position of each particle. The physical origin of the high directionality is attributed to the coherent near-field distribution with large correlation length. This work provides the fundamental theory and physics for future nanoantenna design. PMID- 21633478 TI - Precision analysis in polarization-resolved second harmonic generation microscopy. AB - Polarization-resolved second harmonic generation (PSHG) imaging microscopy allows one to provide information such as anisotropy parameters and molecule orientation. We analyze the precision of parameter estimation for samples with cylindrical symmetry and a Gaussian additive noise model. We introduce improvements of existing techniques that can be useful to get rapid acquisition and processing of PSHG images, and we discuss the influence of photon noise. PMID- 21633479 TI - Studying polymer thin films with hybrid optical microcavities. AB - Organic and inorganic polymeric thin films have numerous applications, including solar cells, biodetection, and nanocomposites. Improving our understanding of the fundamental material behavior is critical to designing polymers with ideal behavior and increased lifetime. However, there are limited nondestructive characterization methods that are able to perform these high-resolution measurements. In this Letter, we demonstrate a method that is able to detect temperature-induced changes in the refractive index of polystyrene polymer thin films as small as 10(-7). This approach is based on optical microcavity resonators. The experimental results agree well with the theoretical simulations. PMID- 21633480 TI - Optical guided waves at graded metal-dielectric interfaces. AB - Waveguides at interfaces with a linearly graded permittivity between a metal and a dielectric are studied. Analytic expressions for the dispersion relations, modes, losses, and cutoff wavelengths are derived and agree well with simulations. The gradation results in anomalous dispersion, a reduction in the energy velocity, and increased field confinement in the metal-dielectric transition region. These effects lead to increased propagation losses, which are sensitive to the spatial extent of the interface gradation. PMID- 21633481 TI - Light self-trapping in a large cloud of cold atoms. AB - We show that, for a near-resonant propagating beam, a large cloud of cold Rb87 atoms acts as a saturable Kerr medium and produces self-trapping of light. By side fluorescence imaging, we monitor the transverse size of the beam and, depending on the sign of the laser detuning with respect to the atomic transition, we observe self-focusing or self-defocusing, with the waist remaining stationary for an appropriate choice of parameters. We analyze our observations by using numerical simulations based on a simple two-level atom model. PMID- 21633482 TI - Formation mechanism of element distribution in glass under femtosecond laser irradiation. AB - We report on the formation mechanism of element distribution in glass under high repetition-rate femtosecond laser irradiation. We simultaneously focused two beams of femtosecond laser pulses inside a glass and confirmed the formation of characteristically shaped element distributions. The results of the numerical simulation in which we considered concentration- and temperature-gradient-driven diffusions were in excellent qualitative agreement with the experimental results, indicating that the main driving force is the sharp temperature gradient. Since the composition of a glass affects its refractive index, absorption, and luminescence property, the results in this study provide a framework to fabricate a functional optical device such as optical circuits with a high-repetition-rate femtosecond laser. PMID- 21633483 TI - Demonstration of a compact 100 Hz, 0.1 J, diode-pumped picosecond laser. AB - We have demonstrated an all-diode-pumped Yb:YAG chirped pulse amplification laser that produces 100 mJ pulses of 5 ps duration at 100 Hz repetition rate. The compact laser system combines a room-temperature Yb:YAG regenerative amplifier for increased bandwidth and a cryogenically cooled Yb:YAG four-pass amplifier for improved heat dissipation and increased efficiency. The optical efficiency of this amplifier is higher than that of other diode-pumped systems of comparable energy. PMID- 21633484 TI - Two-dimensional angular filtering by volume Bragg gratings in photothermorefractive glass. AB - We propose and perform two-dimensional angular filtering on the basis of volume Bragg gratings (VBGs) in a series connection for cleaning up a laser beam. The dependence between angular selectivity and VBG parameters is analyzed with the coupled wave theory. Near-field modulation, contrast ratio, and power spectral density were used to evaluate the effect of filtering in both the spatial and frequency domain. The results from our initial experiments showed that the near field modulation and contrast ratio of laser beam were remarkably improved. PMID- 21633485 TI - Enhanced bandwidth noncollinear optical parametric amplification with a narrowband anamorphic pump. AB - Through the use of anamorphic focusing, we present a method for generating broadband noncollinear optical parametric amplification in signal regions lacking a broadband phase-matching condition that is ideally suited for narrowband pump sources, herein based on an erbium-doped fiber oscillator. With a short focal length cylindrical lens to enhance the phase-matching condition and a long focal length cylindrical lens in the orthogonal plane to limit the pump power in the amplifying beta barium borate crystal, we amplify pulses in the blue-green spectral region with over 100 THz (~3500 cm(-1)) bandwidth. The amplified signal is subsequently compressed to 9.5 fs, near the transform limit. PMID- 21633486 TI - Accurate method for measuring the thermal coefficient of group birefringence of polarization-maintaining fibers. AB - We present a method to accurately measure the group birefringence variation with temperature in high-birefringence polarization-maintaining (PM) fibers using a distributed polarization analyzer. By analyzing polarization cross-talk peaks purposely induced at both ends of a PM fiber, the temperature coefficient of group birefringence can be accurately obtained. We confirm the theoretical prediction that the group birefringence of PANDA and TIGER PM fibers decrease linearly with temperature from -40 degrees C to 80 degrees C, and find that the temperature coefficients are -5.93 * 10(-7) degrees C(-1) and -5.29 * 10(-7) degrees C(-1) for two types of PANDA fibers, and -5.36 * 10(-7) degrees C(-1) for a TIGER fiber. PMID- 21633487 TI - New developments in the pathology of malignant lymphoma: a review of the literature published from October 2009 to January 2010. PMID- 21633488 TI - Interinstitutional and interstate teleneuropathology. AB - BACKGROUND: Telemedicine has emerged as an efficient means of distributing professional medical expertise over a broad geographic area with few limitations to the various services that can be provided around the globe. Telepathology is particularly well suited to distributing subspecialty expertise in certain environments in an economical fashion, while preserving centers of excellence. MATERIALS AND METHODS: After a decade of intrainstitutional teleneuropathology for intraoperative consultation, we expanded our practice to cross state lines and communicate between geographically and financially separate medical centers. RESULTS: The result was an effective means of distributing neuropathological expertise while at the same time preserving a professional center of excellence. While technical and legal (i.e., physician licensing) barriers were surmounted, expected and unexpected issues related to communication required commitment on the part of multiple individuals with diverse expertise and responsibilities. CONCLUSION: Lessons learned from this successful venture can be used to facilitate future efforts in this ever-growing practical vehicle for distributing pathology subspecialty expertise. PMID- 21633489 TI - Standardization in digital pathology: Supplement 145 of the DICOM standards. AB - As digital slides need a lot of storage space, lack of a singular method to acquire and store these large, two-dimensional images has been a major stumbling block in the universal acceptance of this technology. The DICOMS Standard Committee Working Group 26 has put in a tremendous effort to standardize storage methods so that they are more in line with currently available PACS in most hospitals for storage of radiology images. A recent press release (Supplement 145) of these standards was hailed by one and all involved in the field of digital pathology as it will make it easier for hospitals to integrate digital pathology into their already established systems without adding too much overhead costs. Besides, it will enable different vendors developing the scanners to upgrade their products to storage systems that are common across all systems. PMID- 21633490 TI - Reducing patient identification errors related to glucose point-of-care testing. AB - BACKGROUND: Patient identification (ID) errors in point-of-care testing (POCT) can cause test results to be transferred to the wrong patient's chart or prevent results from being transmitted and reported. Despite the implementation of patient barcoding and ongoing operator training at our institution, patient ID errors still occur with glucose POCT. The aim of this study was to develop a solution to reduce identification errors with POCT. MATERIALS AND METHODS: Glucose POCT was performed by approximately 2,400 clinical operators throughout our health system. Patients are identified by scanning in wristband barcodes or by manual data entry using portable glucose meters. Meters are docked to upload data to a database server which then transmits data to any medical record matching the financial number of the test result. With a new model, meters connect to an interface manager where the patient ID (a nine-digit account number) is checked against patient registration data from admission, discharge, and transfer (ADT) feeds and only matched results are transferred to the patient's electronic medical record. With the new process, the patient ID is checked prior to testing, and testing is prevented until ID errors are resolved. RESULTS: When averaged over a period of a month, ID errors were reduced to 3 errors/month (0.015%) in comparison with 61.5 errors/month (0.319%) before implementing the new meters. CONCLUSION: Patient ID errors may occur with glucose POCT despite patient barcoding. The verification of patient identification should ideally take place at the bedside before testing occurs so that the errors can be addressed in real time. The introduction of an ADT feed directly to glucose meters reduced patient ID errors in POCT. PMID- 21633491 TI - Espina: a tool for the automated segmentation and counting of synapses in large stacks of electron microscopy images. AB - The synapses in the cerebral cortex can be classified into two main types, Gray's type I and type II, which correspond to asymmetric (mostly glutamatergic excitatory) and symmetric (inhibitory GABAergic) synapses, respectively. Hence, the quantification and identification of their different types and the proportions in which they are found, is extraordinarily important in terms of brain function. The ideal approach to calculate the number of synapses per unit volume is to analyze 3D samples reconstructed from serial sections. However, obtaining serial sections by transmission electron microscopy is an extremely time consuming and technically demanding task. Using focused ion beam/scanning electron microscope microscopy, we recently showed that virtually all synapses can be accurately identified as asymmetric or symmetric synapses when they are visualized, reconstructed, and quantified from large 3D tissue samples obtained in an automated manner. Nevertheless, the analysis, segmentation, and quantification of synapses is still a labor intensive procedure. Thus, novel solutions are currently necessary to deal with the large volume of data that is being generated by automated 3D electron microscopy. Accordingly, we have developed ESPINA, a software tool that performs the automated segmentation and counting of synapses in a reconstructed 3D volume of the cerebral cortex, and that greatly facilitates and accelerates these processes. PMID- 21633492 TI - Implicit and Explicit Processes in Risk Perception: Neural Antecedents of Perceived HIV Risk. AB - Field studies on HIV risk suggest that people may rely on impressions they have about the safety of their partner at the dispense of more objective risk protection strategies. In this study, ERP recordings were used to investigate the brain mechanisms that give rise to such impressions. First, in an implicit condition, participants viewed a series of photographs of unacquainted persons while performing a task that did not mention HIV risk. Second, in an explicit condition, participants estimated the HIV risk for each presented person. Dense sensor EEG was recorded during the implicit and explicit conditions. In the analysis, explicit risk ratings were used to categorize ERP data from the implicit and explicit conditions into low and high HIV risk categories. The results reveal implicit ERP differences on the basis of subsequent ratings of HIV risk. Specifically, the processing of risky individuals was associated with an early occipital negativity (240-300 ms) and a subsequent central positivity between 430 and 530 ms compared to safe. A similar ERP modulation emerged in the explicit condition for the central positivity component between 430 and 530 ms. A subsequent late positive potential component between 550 and 800 ms was specifically enhanced for risky persons in the explicit rating condition while not modulated in the implicit condition. Furthermore, ratings of HIV risk correlated substantially with ratings of trustworthiness and responsibility. Taken together, these observations provide evidence for theories of intuitive risk perception, which, in the case of HIV risk, seem to operate via appearance based stereotypic inferences. PMID- 21633493 TI - Reward Contingency Modulates Neuronal Activity in Rat Septal Nuclei during Elemental and Configural Association Tasks. AB - It has been suggested that septal nuclei are important in the control of behavior during various reward and non-reward situations. In the present study, neuronal activity was recorded from rat septal nuclei during discrimination of conditioned sensory stimuli (CSs) of the medial forebrain bundle associated with or without a reward (sucrose solution or intracranial self-stimulation, ICSS). Rats were trained to lick a spout protruding close to the mouth just after a CS to obtain a reward stimulus. The CSs included both elemental and configural stimuli. In the configural condition, the reward contingency of the stimuli presented together was opposite to that of each elemental stimulus presented alone, although the same sensory stimuli were involved. Of the 72 responsive septal neurons, 18 responded selectively to the CSs predicting reward (CS(+)-related), four to the CSs predicting non-reward (CS(0)-related), nine to some CSs predicting reward or non-reward, and 15 non-differentially to all CSs. The remaining 26 neurons responded mainly during the ingestion/ICSS phase. A multivariate analysis of the septal neuronal responses to elemental and configural stimuli indicated that septal neurons encoded the CSs based on reward contingency, regardless of the stimulus physical properties and were categorized into three groups; CSs predicting the sucrose solution, CSs predicting a non-reward, and CSs predicting ICSS. The results suggest that septal nuclei are deeply involved in discriminating the reward contingency of environmental stimuli to manifest appropriate behaviors in response to changing stimuli. PMID- 21633494 TI - The immunomodulatory role of syncytiotrophoblast microvesicles. AB - Immune adaptation is a critical component of successful pregnancy. Of primary importance is the modification of cytokine production upon immune activation. With the discovery that normal pregnancy itself is a pro-inflammatory state, it was recognised that the classical Th1/Th2 cytokine paradigm, with a shift towards 'type 2' cytokine production (important for antibody production), and away from 'type 1' immunity (associated with cell mediated immunity and graft rejection), is too simplistic. It is now generally agreed that both arms of cytokine immunity are activated, but with a bias towards 'type 2' immunity. Many factors are released from the placenta that can influence the maternal cytokine balance. Here we focus on syncytiotrophoblast microvesicles (STBM) which are shed from the placenta into the maternal circulation. We show that STBM can bind to monocytes and B cells and induce cytokine release (TNFalpha, MIP-1alpha, IL-1alpha, IL 1beta, IL-6, IL-8). Other cytokines are down-modulated, such as IP-10 which is associated with 'type 1' immunity. Therefore STBM may aid the 'type 2' skewed nature of normal pregnancy. We also observed that PBMC from third trimester normal pregnant women produce more TNFalpha and IL-6 in response to STBM than PBMC from non-pregnant women, confirming that maternal immune cells are primed by pregnancy, possibly through their interaction with STBM. PMID- 21633495 TI - Using workflows to explore and optimise named entity recognition for chemistry. AB - Chemistry text mining tools should be interoperable and adaptable regardless of system-level implementation, installation or even programming issues. We aim to abstract the functionality of these tools from the underlying implementation via reconfigurable workflows for automatically identifying chemical names. To achieve this, we refactored an established named entity recogniser (in the chemistry domain), OSCAR and studied the impact of each component on the net performance. We developed two reconfigurable workflows from OSCAR using an interoperable text mining framework, U-Compare. These workflows can be altered using the drag-&-drop mechanism of the graphical user interface of U-Compare. These workflows also provide a platform to study the relationship between text mining components such as tokenisation and named entity recognition (using maximum entropy Markov model (MEMM) and pattern recognition based classifiers). Results indicate that, for chemistry in particular, eliminating noise generated by tokenisation techniques lead to a slightly better performance than others, in terms of named entity recognition (NER) accuracy. Poor tokenisation translates into poorer input to the classifier components which in turn leads to an increase in Type I or Type II errors, thus, lowering the overall performance. On the Sciborg corpus, the workflow based system, which uses a new tokeniser whilst retaining the same MEMM component, increases the F-score from 82.35% to 84.44%. On the PubMed corpus, it recorded an F-score of 84.84% as against 84.23% by OSCAR. PMID- 21633496 TI - Neural correlates of natural human echolocation in early and late blind echolocation experts. AB - BACKGROUND: A small number of blind people are adept at echolocating silent objects simply by producing mouth clicks and listening to the returning echoes. Yet the neural architecture underlying this type of aid-free human echolocation has not been investigated. To tackle this question, we recruited echolocation experts, one early- and one late-blind, and measured functional brain activity in each of them while they listened to their own echolocation sounds. RESULTS: When we compared brain activity for sounds that contained both clicks and the returning echoes with brain activity for control sounds that did not contain the echoes, but were otherwise acoustically matched, we found activity in calcarine cortex in both individuals. Importantly, for the same comparison, we did not observe a difference in activity in auditory cortex. In the early-blind, but not the late-blind participant, we also found that the calcarine activity was greater for echoes reflected from surfaces located in contralateral space. Finally, in both individuals, we found activation in middle temporal and nearby cortical regions when they listened to echoes reflected from moving targets. CONCLUSIONS: These findings suggest that processing of click-echoes recruits brain regions typically devoted to vision rather than audition in both early and late blind echolocation experts. PMID- 21633497 TI - Novel insights into the diversity of catabolic metabolism from ten haloarchaeal genomes. AB - BACKGROUND: The extremely halophilic archaea are present worldwide in saline environments and have important biotechnological applications. Ten complete genomes of haloarchaea are now available, providing an opportunity for comparative analysis. METHODOLOGY/PRINCIPAL FINDINGS: We report here the comparative analysis of five newly sequenced haloarchaeal genomes with five previously published ones. Whole genome trees based on protein sequences provide strong support for deep relationships between the ten organisms. Using a soft clustering approach, we identified 887 protein clusters present in all halophiles. Of these core clusters, 112 are not found in any other archaea and therefore constitute the haloarchaeal signature. Four of the halophiles were isolated from water, and four were isolated from soil or sediment. Although there are few habitat-specific clusters, the soil/sediment halophiles tend to have greater capacity for polysaccharide degradation, siderophore synthesis, and cell wall modification. Halorhabdus utahensis and Haloterrigena turkmenica encode over forty glycosyl hydrolases each, and may be capable of breaking down naturally occurring complex carbohydrates. H. utahensis is specialized for growth on carbohydrates and has few amino acid degradation pathways. It uses the non oxidative pentose phosphate pathway instead of the oxidative pathway, giving it more flexibility in the metabolism of pentoses. CONCLUSIONS: These new genomes expand our understanding of haloarchaeal catabolic pathways, providing a basis for further experimental analysis, especially with regard to carbohydrate metabolism. Halophilic glycosyl hydrolases for use in biofuel production are more likely to be found in halophiles isolated from soil or sediment. PMID- 21633498 TI - Disease and the extended phenotype: parasites control host performance and survival through induced changes in body plan. AB - BACKGROUND: By definition, parasites harm their hosts. However, some forms of parasite-induced alterations increase parasite transmission between hosts, such that manipulated hosts can be considered extensions of the parasite's phenotype. While well accepted in principle, surprisingly few studies have quantified how parasite manipulations alter host performance and survival under field and laboratory conditions. METHODOLOGY/PRINCIPAL FINDINGS: By interfering with limb development, the trematode Ribeiroia ondatrae causes particularly severe morphological alterations within amphibian hosts that provide an ideal system to evaluate parasite-induced changes in phenotype. Here, we coupled laboratory performance trials with a capture-mark-recapture study of 1388 Pacific chorus frogs (Pseudacris regilla) to quantify the effects of parasite-induced malformations on host locomotion, foraging, and survival. Malformations, which affected ~ 50% of metamorphosing frogs in nature, caused dramatic reductions in all measures of organismal function. Malformed frogs exhibited significantly shorter jumping distances (41% reduction), slower swimming speeds (37% reduction), reduced endurance (66% reduction), and lower foraging success relative to infected hosts without malformations. Furthermore, while normal and malformed individuals had comparable survival within predator-free exclosures, deformed frogs in natural populations had 22% lower biweekly survival than normal frogs and rarely recruited to the adult population over a two-year period. CONCLUSIONS/SIGNIFICANCE: Our results highlight the ability of parasites to deeply alter multiple dimensions of host phenotype with important consequences for performance and survival. These patterns were best explained by malformation status, rather than infection per se, helping to decouple the direct and indirect effects of parasitism on host fitness. PMID- 21633499 TI - Perception of male caller identity in Koalas (Phascolarctos cinereus): acoustic analysis and playback experiments. AB - The ability to signal individual identity using vocal signals and distinguish between conspecifics based on vocal cues is important in several mammal species. Furthermore, it can be important for receivers to differentiate between callers in reproductive contexts. In this study, we used acoustic analyses to determine whether male koala bellows are individually distinctive and to investigate the relative importance of different acoustic features for coding individuality. We then used a habituation-discrimination paradigm to investigate whether koalas discriminate between the bellow vocalisations of different male callers. Our results show that male koala bellows are highly individualized, and indicate that cues related to vocal tract filtering contribute the most to vocal identity. In addition, we found that male and female koalas habituated to the bellows of a specific male showed a significant dishabituation when they were presented with bellows from a novel male. The significant reduction in behavioural response to a final rehabituation playback shows this was not a chance rebound in response levels. Our findings indicate that male koala bellows are highly individually distinctive and that the identity of male callers is functionally relevant to male and female koalas during the breeding season. We go on to discuss the biological relevance of signalling identity in this species' sexual communication and the potential practical implications of our findings for acoustic monitoring of male population levels. PMID- 21633500 TI - Interaction between amyloid beta peptide and an aggregation blocker peptide mimicking islet amyloid polypeptide. AB - Assembly of amyloid-beta peptide (Abeta) into cytotoxic oligomeric and fibrillar aggregates is believed to be a major pathologic event in Alzheimer's disease (AD) and interfering with Abeta aggregation is an important strategy in the development of novel therapeutic approaches. Prior studies have shown that the double N-methylated analogue of islet amyloid polypeptide (IAPP) IAPP-GI, which is a conformationally constrained IAPP analogue mimicking a non-amyloidogenic IAPP conformation, is capable of blocking cytotoxic self-assembly of Abeta. Here we investigate the interaction of IAPP-GI with Abeta40 and Abeta42 using NMR spectroscopy. The most pronounced NMR chemical shift changes were observed for residues 13-20, while residues 7-9, 15-16 as well as the C-terminal half of Abeta -that is both regions of the Abeta sequence that are converted into beta-strands in amyloid fibrils--were less accessible to solvent in the presence of IAPP-GI. At the same time, interaction of IAPP-GI with Abeta resulted in a concentration dependent co-aggregation of Abeta and IAPP-GI that was enhanced for the more aggregation prone Abeta42 peptide. On the basis of the reduced toxicity of the Abeta peptide in the presence of IAPP-GI, our data are consistent with the suggestion that IAPP-GI redirects Abeta into nontoxic "off-pathway" aggregates. PMID- 21633501 TI - Reevaluation of the value of autoparasitoids in biological control. AB - Autoparasitoids with the capacity of consuming primary parasitoids that share the same hosts to produce males are analogous to intraguild predators. The use of autoparasitoids in biological control programs is a controversial matter because there is little evidence to support the view that autoparasitoids do not disrupt and at times may promote suppression of insect pests in combination with primary parasitoids. We found that Encarsia sophia, a facultative autoparasitoid, preferred to use heterospecific hosts as secondary hosts for producing males. The autoparasitoids mated with males originated from heterospecifics may parasitize more hosts than those mated with males from conspecifics. Provided with an adequate number of males, the autoparasitoids killed more hosts than En. formosa, a commonly used parasitoid for biological control of whiteflies. This study supports the view that autoparasitoids in combination with primary parasitoids do not disrupt pest management and may enhance such programs. The demonstrated preference of an autoparasitoid for heterospecifics and improved performance of males from heterospecifics observed in this study suggests these criteria should be considered in strategies that endeavor to mass-produce and utilize autoparasitoids in the future. PMID- 21633502 TI - Adipocytokines and CD34 progenitor cells in Alzheimer's disease. AB - BACKGROUND: Alzheimer's disease (AD) and atherosclerosis share common vascular risk factors such as arterial hypertension and hypercholesterolemia. Adipocytokines and CD34(+) progenitor cells are associated with the progression and prognosis of atherosclerotic diseases. Their role in AD is not adequately elucidated. METHODS AND FINDINGS: In the present study, we measured in 41 patients with early AD and 37 age- and weight-matched healthy controls blood concentrations of adiponectin and leptin by enzyme linked immunoabsorbent assay and of CD34(+) progenitor cells using flow cytometry. We found significantly lower plasma levels of leptin in AD patients compared with the controls, whereas plasma levels of adiponectin did not show any significant differences (AD vs. control (mean +/- SD): leptin:8.9 +/- 5.6 ng/mL vs.16.3 +/- 15.5 ng/mL;P = 0.038; adiponectin:18.5 +/- 18.1 ug/mL vs.16.7 +/- 8.9 ug/mL;P = 0.641). In contrast, circulating CD34(+) cells were significantly upregulated in AD patients (mean absolute cell count +/- SD:253 +/- 51 vs. 203 +/- 37; P = 0.02) and showed an inverse correlation with plasma levels of leptin (r = -0.248; P = 0.037). In logistic regression analysis, decreased leptin concentration (P = 0.021) and increased number of CD34(+) cells (P = 0.036) were both significantly associated with the presence of AD. According to multifactorial analysis of covariance, leptin serum levels were a significant independent predictor for the number of CD34(+) cells (P = 0.002). CONCLUSIONS: Our findings suggest that low plasma levels of leptin and increased numbers of CD34(+) progenitor cells are both associated with AD. In addition, the results of our study provide first evidence that increased leptin plasma levels are associated with a reduced number of CD34(+) progenitor cells in AD patients. These findings point towards a combined involvement of leptin and CD34(+) progenitor cells in the pathogenesis of AD. Thus, plasma levels of leptin and circulating CD34(+) progenitor cells could represent an important molecular link between atherosclerotic diseases and AD. Further studies should clarify the pathophysiological role of both adipocytokines and progenitor cells in AD and possible diagnostic and therapeutic applications. PMID- 21633503 TI - Being Barbie: the size of one's own body determines the perceived size of the world. AB - A classical question in philosophy and psychology is if the sense of one's body influences how one visually perceives the world. Several theoreticians have suggested that our own body serves as a fundamental reference in visual perception of sizes and distances, although compelling experimental evidence for this hypothesis is lacking. In contrast, modern textbooks typically explain the perception of object size and distance by the combination of information from different visual cues. Here, we describe full body illusions in which subjects experience the ownership of a doll's body (80 cm or 30 cm) and a giant's body (400 cm) and use these as tools to demonstrate that the size of one's sensed own body directly influences the perception of object size and distance. These effects were quantified in ten separate experiments with complementary verbal, questionnaire, manual, walking, and physiological measures. When participants experienced the tiny body as their own, they perceived objects to be larger and farther away, and when they experienced the large-body illusion, they perceived objects to be smaller and nearer. Importantly, despite identical retinal input, this "body size effect" was greater when the participants experienced a sense of ownership of the artificial bodies compared to a control condition in which ownership was disrupted. These findings are fundamentally important as they suggest a causal relationship between the representations of body space and external space. Thus, our own body size affects how we perceive the world. PMID- 21633504 TI - In-silico patterning of vascular mesenchymal cells in three dimensions. AB - Cells organize in complex three-dimensional patterns by interacting with proteins along with the surrounding extracellular matrix. This organization provides the mechanical and chemical cues that ultimately influence a cell's differentiation and function. Here, we computationally investigate the pattern formation process of vascular mesenchymal cells arising from their interaction with Bone Morphogenic Protein-2 (BMP-2) and its inhibitor, Matrix Gla Protein (MGP). Using a first-principles approach, we derive a reaction-diffusion model based on the biochemical interactions of BMP-2, MGP and cells. Simulations of the model exhibit a wide variety of three-dimensional patterns not observed in a two dimensional analysis. We demonstrate the emergence of three types of patterns: spheres, tubes, and sheets, and show that the patterns can be tuned by modifying parameters in the model such as the degradation rates of proteins and chemotactic coefficient of cells. Our model may be useful for improved engineering of three dimensional tissue structures as well as for understanding three dimensional microenvironments in developmental processes. PMID- 21633505 TI - Neutralising antibodies against ricin toxin. AB - The Centers for Disease Control and Prevention have listed the potential bioweapon ricin as a Category B Agent. Ricin is a so-called A/B toxin produced by plants and is one of the deadliest molecules known. It is easy to prepare and no curative treatment is available. An immunotherapeutic approach could be of interest to attenuate or neutralise the effects of the toxin. We sought to characterise neutralising monoclonal antibodies against ricin and to develop an effective therapy. For this purpose, mouse monoclonal antibodies (mAbs) were produced against the two chains of ricin toxin (RTA and RTB). Seven mAbs were selected for their capacity to neutralise the cytotoxic effects of ricin in vitro. Three of these, two anti-RTB (RB34 and RB37) and one anti-RTA (RA36), when used in combination improved neutralising capacity in vitro with an IC(50) of 31 ng/ml. Passive administration of association of these three mixed mAbs (4.7 ug) protected mice from intranasal challenges with ricin (5 LD(50)). Among those three antibodies, anti-RTB antibodies protected mice more efficiently than the anti-RTA antibody. The combination of the three antibodies protected mice up to 7.5 hours after ricin challenge. The strong in vivo neutralising capacity of this three mAbs combination makes it potentially useful for immunotherapeutic purposes in the case of ricin poisoning or possibly for prevention. PMID- 21633506 TI - Do psychological variables affect early surgical recovery? AB - BACKGROUND: Numerous studies have examined the effect of psychological variables on surgical recovery, but no definite conclusion has been reached yet. We sought to examine whether psychological factors influence early surgical recovery. METHODS: We performed a systematic search in PubMed, Scopus and PsycINFO databases to identify studies examining the association of preoperative psychological variables or interventions with objectively measured, early surgical outcomes. RESULTS: We identified 16 eligible studies, 15 of which reported a significant association between at least one psychological variable or intervention and an early postoperative outcome. However, most studies also reported psychological factors not influencing surgical recovery and there was significant heterogeneity across the studies. Overall, trait and state anxiety, state anger, active coping, subclinical depression, and intramarital hostility appeared to complicate recovery, while dispositional optimism, religiousness, anger control, low pain expectations, and external locus of control seemed to promote healing. Psychological interventions (guided relaxation, couple support visit, and psychiatric interview) also appeared to favor recovery. Psychological factors unrelated to surgical outcomes included loneliness, perceived social support, anger expression, and trait anger. CONCLUSION: Although the heterogeneity of the available evidence precludes any safe conclusions, psychological variables appear to be associated with early surgical recovery; this association could bear important implications for clinical practice. Large clinical trials and further analyses are needed to precisely evaluate the contribution of psychology in surgical recovery. PMID- 21633507 TI - The intestinal microbiota plays a role in Salmonella-induced colitis independent of pathogen colonization. AB - The intestinal microbiota is composed of hundreds of species of bacteria, fungi and protozoa and is critical for numerous biological processes, such as nutrient acquisition, vitamin production, and colonization resistance against bacterial pathogens. We studied the role of the intestinal microbiota on host resistance to Salmonella enterica serovar Typhimurium-induced colitis. Using multiple antibiotic treatments in 129S1/SvImJ mice, we showed that disruption of the intestinal microbiota alters host susceptibility to infection. Although all antibiotic treatments caused similar increases in pathogen colonization, the development of enterocolitis was seen only when streptomycin or vancomycin was used; no significant pathology was observed with the use of metronidazole. Interestingly, metronidazole-treated and infected C57BL/6 mice developed severe pathology. We hypothesized that the intestinal microbiota confers resistance to infectious colitis without affecting the ability of S. Typhimurium to colonize the intestine. Indeed, different antibiotic treatments caused distinct shifts in the intestinal microbiota prior to infection. Through fluorescence in situ hybridization, terminal restriction fragment length polymorphism, and real-time PCR, we showed that there is a strong correlation between the intestinal microbiota composition before infection and susceptibility to Salmonella-induced colitis. Members of the Bacteroidetes phylum were present at significantly higher levels in mice resistant to colitis. Further analysis revealed that Porphyromonadaceae levels were also increased in these mice. Conversely, there was a positive correlation between the abundance of Lactobacillus sp. and predisposition to colitis. Our data suggests that different members of the microbiota might be associated with S. Typhimurium colonization and colitis. Dissecting the mechanisms involved in resistance to infection and inflammation will be critical for the development of therapeutic and preventative measures against enteric pathogens. PMID- 21633508 TI - GLI1 confers profound phenotypic changes upon LNCaP prostate cancer cells that include the acquisition of a hormone independent state. AB - The GLI (GLI1/GLI2) transcription factors have been implicated in the development and progression of prostate cancer although our understanding of how they actually contribute to the biology of these common tumours is limited. We observed that GLI reporter activity was higher in normal (PNT-2) and tumourigenic (DU145 and PC-3) androgen-independent cells compared to androgen-dependent LNCaP prostate cancer cells and, accordingly, GLI mRNA levels were also elevated. Ectopic expression of GLI1 or the constitutively active DeltaNGLI2 mutant induced a distinct cobblestone-like morphology in LNCaP cells that, regarding the former, correlated with increased GLI2 as well as expression of the basal/stem-like markers CD44, beta1-integrin, DeltaNp63 and BMI1, and decreased expression of the luminal marker AR (androgen receptor). LNCaP-GLI1 cells were viable in the presence of the AR inhibitor bicalutamide and gene expression profiling revealed that the transcriptome of LNCaP-GLI1 cells was significantly closer to DU145 and PC-3 cells than to control LNCaP-pBP (empty vector) cells, as well as identifying LCN2/NGAL as a highly induced transcript which is associated with hormone independence in breast and prostate cancer. Functionally, LNCaP-GLI1 cells displayed greater clonal growth and were more invasive than control cells but they did not form colonies in soft agar or prostaspheres in suspension suggesting that they do not possess inherent stem cell properties. Moreover, targeted suppression of GLI1 or GLI2 with siRNA did not reverse the transformed phenotype of LNCaP-GLI1 cells nor did double GLI1/GLI2 knockdowns activate AR expression in DU145 or PC-3 cells. As such, early targeting of the GLI oncoproteins may hinder progression to a hormone independent state but a more detailed understanding of the mechanisms that maintain this phenotype is required to determine if their inhibition will enhance the efficacy of anti-hormonal therapy through the induction of a luminal phenotype and increased dependency upon AR function. PMID- 21633509 TI - Diversity of prophage DNA regions of Streptococcus agalactiae clonal lineages from adults and neonates with invasive infectious disease. AB - The phylogenetic position and prophage DNA content of the genomes of 142 S. agalactiae (group-B streptococcus, GBS) isolates responsible for bacteremia and meningitis in adults and neonates were studied and compared. The distribution of the invasive isolates between the various serotypes, sequence types (STs) and clonal complexes (CCs) differed significantly between adult and neonatal isolates. Use of the neighbor-net algorithm with the PHI test revealed evidence for recombination in the population studied (PHI, P = 2.01 * 10(-6)), and the recombination-mutation ratio (R/M) was 6:7. Nevertheless, the estimated R/M ratio differed between CCs. Analysis of the prophage DNA regions of the genomes of the isolates assigned 90% of the isolates to five major prophage DNA groups: A to E. The mean number of prophage DNA fragments amplified per isolate varied from 2.6 for the isolates of prophage DNA group E to 4.0 for the isolates of prophage DNA group C. The isolates from adults and neonates with invasive diseases were distributed differently between the various prophage DNA groups (P < 0.00001). Group C prophage DNA fragments were found in 52% of adult invasive isolates, whereas 74% of neonatal invasive isolates had prophage DNA fragments of groups A and B. Differences in prophage DNA content were also found between serotypes, STs and CCs (P < 0.00001). All the ST-1 and CC1 isolates, mostly of serotype V, belonged to the prophage DNA group C, whereas 84% of the ST-17 and CC17 isolates, all of serotype III, belonged to prophage DNA groups A and B. These data indicate that the transduction mechanisms, i.e., gene transfer from one bacterium to another by a bacteriophage, underlying genetic recombination in S. agalactiae species, are specific to each intraspecies lineage and population of strains responsible for invasive diseases in adults and neonates. PMID- 21633510 TI - Augmentation cystoplasty and extracellular matrix scaffolds: an ex vivo comparative study with autogenous detubularised ileum. AB - BACKGROUND: Augmentation cystoplasty (AC) with autogenous ileum remains the current gold standard surgical treatment for many patients with end-stage bladder disease. However, the presence of mucus-secreting epithelium within the bladder is associated with debilitating long-term complications. Currently, decellularised biological materials derived from porcine extracellular matrix (ECM) are under investigation as potential augmentation scaffolds. Important biomechanical limitations of ECMs are decreased bladder capacity and poor compliance after implantation. METHODOLOGY/PRINCIPAL FINDINGS: In the present ex vivo study a novel concept was investigated where a two-fold increase in ECM scaffold surface-area relative to the resected ileal segment was compared in ovine bladder models after AC. Results showed that bladder capacity increased by 40 +/- 4% and 37 +/- 11% at 10 mmHg and compliance by 40.4 +/- 4% and 39.7 +/- 6% (DeltaP = 0-10 mmHg) after AC with ileum and porcine urinary bladder matrix (UBM) respectively (p < 0.05). Comparative assessment between ileum and UBM demonstrated no significant differences in bladder capacity or compliance increases after AC (p > 0.05). CONCLUSIONS: These findings may have important clinical implications as metabolic, infective and malignant complications precipitated by mucus-secreting epithelium are potentially avoided after augmentation with ECM scaffolds. PMID- 21633511 TI - Human endogenous retrovirus K106 (HERV-K106) was infectious after the emergence of anatomically modern humans. AB - HERV-K113 and HERV-K115 have been considered to be among the youngest HERVs because they are the only known full-length proviruses that are insertionally polymorphic and maintain the open reading frames of their coding genes. However, recent data suggest that HERV-K113 is at least 800,000 years old, and HERV-K115 even older. A systematic study of HERV-K HML2 members to identify HERVs that may have infected the human genome in the more recent evolutionary past is lacking. Therefore, we sought to determine how recently HERVs were exogenous and infectious by examining sequence variation in the long terminal repeat (LTR) regions of all full-length HERV-K loci. We used the traditional method of inter LTR comparison to analyze all full length HERV-Ks and determined that two insertions, HERV-K106 and HERV-K116 have no differences between their 5' and 3' LTR sequences, suggesting that these insertions were endogenized in the recent evolutionary past. Among these insertions with no sequence differences between their LTR regions, HERV-K106 had the most intact viral sequence structure. Coalescent analysis of HERV-K106 3' LTR sequences representing 51 ethnically diverse individuals suggests that HERV-K106 integrated into the human germ line approximately 150,000 years ago, after the emergence of anatomically modern humans. PMID- 21633512 TI - New ABA-hypersensitive Arabidopsis mutants are affected in loci mediating responses to water deficit and Dickeya dadantii infection. AB - On water deficit, abscisic acid (ABA) induces stomata closure to reduce water loss by transpiration. To identify Arabidopsis thaliana mutants which transpire less on drought, infrared thermal imaging of leaf temperature has been used to screen for suppressors of an ABA-deficient mutant (aba3-1) cold-leaf phenotype. Three novel mutants, called hot ABA-deficiency suppressor (has), have been identified with hot-leaf phenotypes in the absence of the aba3 mutation. The defective genes imparted no apparent modification to ABA production on water deficit, were inherited recessively and enhanced ABA responses indicating that the proteins encoded are negative regulators of ABA signalling. All three mutants showed ABA-hypersensitive stomata closure and inhibition of root elongation with little modification of growth and development in non-stressed conditions. The has2 mutant also exhibited increased germination inhibition by ABA, while ABA inducible gene expression was not modified on dehydration, indicating the mutated gene affects early ABA-signalling responses that do not modify transcript levels. In contrast, weak ABA-hypersensitivity relative to mutant developmental phenotypes suggests that HAS3 regulates drought responses by both ABA-dependent and independent pathways. has1 mutant phenotypes were only apparent on stress or ABA treatments, and included reduced water loss on rapid dehydration. The HAS1 locus thus has the required characteristics for a targeted approach to improving resistance to water deficit. In contrast to has2, has1 exhibited only minor changes in susceptibility to Dickeya dadantii despite similar ABA hypersensitivity, indicating that crosstalk between ABA responses to this pathogen and drought stress can occur through more than one point in the signalling pathway. PMID- 21633513 TI - Novel polymorphisms in Plasmodium falciparum ABC transporter genes are associated with major ACT antimalarial drug resistance. AB - Chemotherapy is a critical component of malaria control. However, the most deadly malaria pathogen, Plasmodium falciparum, has repeatedly mounted resistance against a series of antimalarial drugs used in the last decades. Southeast Asia is an epicenter of emerging antimalarial drug resistance, including recent resistance to the artemisinins, the core component of all recommended antimalarial combination therapies. Alterations in the parasitic membrane proteins Pgh-1, PfCRT and PfMRP1 are believed to be major contributors to resistance through decreasing intracellular drug accumulation. The pfcrt, pfmdr1 and pfmrp1 genes were sequenced from a set of P.falciparum field isolates from the Thai-Myanmar border. In vitro drug susceptibility to artemisinin, dihydroartemisinin, mefloquine and lumefantrine were assessed. Positive correlations were seen between the in vitro susceptibility responses to artemisinin and dihydroartemisinin and the responses to the arylamino-alcohol quinolines lumefantrine and mefloquine. The previously unstudied pfmdr1 F1226Y and pfmrp1 F1390I SNPs were associated significantly with artemisinin, mefloquine and lumefantrine in vitro susceptibility. A variation in pfmdr1 gene copy number was also associated with parasite drug susceptibility of artemisinin, mefloquine and lumefantrine. Our work unveils new candidate markers of P. falciparum multidrug resistance in vitro, while contributing to the understanding of subjacent genetic complexity, essential for future evidence-based drug policy decisions. PMID- 21633514 TI - Cost-effectiveness of early treatment with first-line NNRTI-based HAART regimens in the UK, 1996-2006. AB - AIM: Calculate time to first-line treatment failure, annual cost and cost effectiveness of NNRTI versus PIboosted first-line HAART regimens in the UK, 1996 2006. BACKGROUND: Population costs for HIV services are increasing in the UK and interventions need to be effective and efficient to reduce or stabilize costs. 2NRTIs + NNRTI regimens are cost-effective regimens for first-line HAART, but these regimens have not been compared with first-line PI(boosted) regimens. METHODS: Times to first-line treatment failure and annual costs were calculated for first-line HAART regimens by CD4 count when starting HAART (2006 UK prices). Cost-effectiveness of 2NRTIs+NNRTI versus 2NRTIs+PI(boosted) regimens was calculated for four CD4 strata. RESULTS: 55% of 5,541 people living with HIV (PLHIV) started HAART with CD4 count <= 200 cells/mm3, many of whom were Black Africans. Annual treatment cost decreased as CD4 count increased; most marked differences were observed between starting HAART with CD4 <= 200 cells/mm3 compared with CD4 count >200 cells/mm3. 2NRTI+PI(boosted) and 2NRTI+NNRTI regimens were the most effective regimens across the four CD4 strata; 2NRTI + NNRTI was cost-saving or cost-effective compared with 2NRTI + PI(boosted) regimens. CONCLUSION: To ensure more effective and efficient provision of HIV services, 2NRTI+NNRTI should be started as first-line HAART regimen at CD4 counts <= 350 cell/mm3, unless specific contra-indications exist. This will increase the number of PLHIV receiving HAART and will initially increase population costs of providing HIV services. However, starting PLHIV earlier on cost-effective regimens will maintain them in better health and use fewer health or social services, thereby generating fewer treatment and care costs, enabling them to remain socially and economically active members of society. This does raise a number of ethical issues, which will have to be acknowledged and addressed, especially in countries with limited resources. PMID- 21633515 TI - Visual information alone changes behavior and physiology during social interactions in a cichlid fish (Astatotilapia burtoni). AB - Social behavior can influence physiological systems dramatically yet the sensory cues responsible are not well understood. Behavior of male African cichlid fish, Astatotilapia burtoni, in their natural habitat suggests that visual cues from conspecifics contribute significantly to regulation of social behavior. Using a novel paradigm, we asked whether visual cues alone from a larger conspecific male could influence behavior, reproductive physiology and the physiological stress response of a smaller male. Here we show that just seeing a larger, threatening male through a clear barrier can suppress dominant behavior of a smaller male for up to 7 days. Smaller dominant males being "attacked" visually by larger dominant males through a clear barrier also showed physiological changes for up to 3 days, including up-regulation of reproductive- and stress-related gene expression levels and lowered plasma 11-ketotestesterone concentrations as compared to control animals. The smaller males modified their appearance to match that of non dominant males when exposed to a larger male but they maintained a physiological phenotype similar to that of a dominant male. After 7 days, reproductive- and stress- related gene expression, circulating hormone levels, and gonad size in the smaller males showed no difference from the control group suggesting that the smaller male habituated to the visual intruder. However, the smaller male continued to display subordinate behaviors and assumed the appearance of a subordinate male for a full week despite his dominant male physiology. These data suggest that seeing a larger male alone can regulate the behavior of a smaller male but that ongoing reproductive inhibition depends on additional sensory cues. Perhaps, while experiencing visual social stressors, the smaller male uses an opportunistic strategy, acting like a subordinate male while maintaining the physiology of a dominant male. PMID- 21633516 TI - Negligible contribution from roots to soil-borne phospholipid fatty acid fungal biomarkers 18:2omega6,9 and 18:1omega9. AB - The phospholipid fatty acid biomarkers 18:1omega9, 18:2omega6,9 and 18:3omega3,6,9 are commonly used as fungal biomarkers in soils. They have, however, also been found to occur in plant tissues, such as roots. Thus, the use of these PLFAs as fungal biomarkers in sieved soil, which may still contain small remains of roots, has been questioned. We used data from a recent beech tree girdling experiment to calculate the contribution of roots to these biomarkers and were able to demonstrate that not more than 0.61% of 18:1omega9 and 18:2omega6,9 in sieved soil samples originated from roots (but 4% of 18:3omega3,6,9). Additionally, the abundance of the biomarker 18:2omega6,9 in the soil was found to be highly correlated to ectomycorrhizal root colonization, which further corroborates its fungal origin. PLFA biomarkers were substantially reduced in vital roots from girdled trees compared to roots of control trees (by up to 76%), indicating that the major part of PLFAs measured in roots may actually originate from ectomycorrhizal fungi growing inside the roots. We calculated, that even a near to 50% reduction in fine root biomass - as observed in the girdling treatment - accounted for only 0.8% of the measured decrease of 18:2omega6,9. Our results demonstrate that both 18:1omega9 and 18:2omega6,9 are suitable biomarkers for detecting fungal dynamics in soils and that especially 18:2omega6,9 is a reliable biomarker to study mycorrhizal dynamics in beech forests. PMID- 21633517 TI - On the 'temperature sensitivity' of soil respiration: Can we use the immeasurable to predict the unknown? AB - The temperature dependence of soil respiration (R(S)) is widely used as a key characteristic of soils or organic matter fractions within soils, and in the context of global climatic change is often applied to infer likely responses of R(S) to warmer future conditions. However, the way in which these temperature dependencies are calculated, interpreted and implemented in ecosystem models requires careful consideration of possible artefacts and assumptions. We argue that more conceptual clarity in the reported relationships is needed to obtain meaningful meta-analyses and better constrained parameters informing ecosystem models. Our critical assessment of common methodologies shows that it is impossible to measure actual temperature response of R(S), and that a range of confounding effects creates the observed apparent temperature relations reported in the literature. Thus, any measureable temperature response function will likely fail to predict effects of climate change on R(s). For improving our understanding of R(S) in changing environments we need a better integration of the relationships between substrate supply and the soil biota, and of their long term responses to changes in abiotic soil conditions. This is best achieved by experiments combining isotopic techniques and ecosystem manipulations, which allow a disentangling of abiotic and biotic factors underlying the temperature response of soil CO(2) efflux. PMID- 21633518 TI - Your own actions influence how you perceive other people: A misattribution of action appraisals. AB - The attribution of personal traits to other persons depends on the actions the observer performs at the same time (Bach & Tipper, 2007). Here, we show that the effect reflects a misattribution of appraisals of the observers' own actions to the actions of others. We exploited spatial compatibility effects to manipulate how fluently-how fast and how accurately-participants identified two individuals performing sporty or academic actions. The traits attributed to each person in a subsequent rating task depended on the fluency of participants' responses in a specific manner. An individual more fluently identified while performing the academic action appeared more academic and less sporty. An individual more fluently identified while performing the sporty action appeared sportier. Thus, social perception is-at least partially-embodied. The ease of our own responses can be misattributed to the actions of others, affecting which personal traits are attributed to them. PMID- 21633519 TI - First measurement of kaonic helium-3 X-rays. AB - The first observation of the kaonic (3)He 3d->2p transition was made, using slow K- mesons stopped in a gaseous (3)He target. The kaonic atom X-rays were detected with large-area silicon drift detectors using the timing information of the K+K- pairs of phi-meson decays produced by the DAPhiNE e+e- collider. The strong interaction shift of the kaonic (3)He 2p state was determined to be -2+/ 2(stat)+/-4(syst) eV. PMID- 21633520 TI - Cascade Screening for Familial Hypercholesterolemia (FH). AB - Familial hypercholesterolemia (FH) is an autosomal dominant disorder characterized by abnormally high concentrations of low-density lipoprotein (LDL) cholesterol in the blood, which predisposes affected persons to premature coronary heart disease (CHD) and death. FH is one of the most common inherited disorders and the most common one known to cause premature CHD in people of European descent. The vast majority of people with FH have inherited a single mutation from one parent in either the LDL receptor (LDLR), apolipoprotein B (APOB), or proprotein convertase subtilisin/kexin type 9 (PCSK9) genes. Despite their greatly elevated risk of coronary heart disease, most individuals with FH remain undiagnosed, untreated, or inadequately treated. Cascade screening is a mechanism for identifying people at risk for a genetic condition by a process of systematic family tracing. The National Institute for Health and Clinical Excellence in the United Kingdom recommends cascade screening of close biological relatives of people with a clinical diagnosis of FH in order to effectively identify additional FH patients. The ultimate goal of this testing is to reduce morbidity and mortality from heart disease in persons with FH through early diagnosis and effective disease management. The goal of this article is to outline the available evidence on the clinical validity and utility of cascade screening for FH, while emphasizing the availability, usefulness, and recommendation for including DNA testing (if the disease-causing mutation has been identified). PMID- 21633521 TI - Reappraisal of role of angiotensin receptor blockers in cardiovascular protection. AB - Angiotensin-converting enzyme inhibitors (ACEIs) and angiotensin receptor blockers (ARBs) have shown cardioprotective and renoprotective properties. These agents are recommended as first-line therapy for the treatment of hypertension and the reduction of cardiovascular risk. Early studies pointed to the cardioprotective and renoprotective effects of ARBs in high-risk patients. The ONgoing Telmisartan Alone and in combination with Ramipril Global Endpoint Trial (ONTARGET) established the clinical equivalence of the cardioprotective and renoprotective effects of telmisartan and ramipril, but did not find an added benefit of the combination over ramipril alone. Similar findings were observed in the Telmisartan Randomized AssessmeNt Study in aCE INtolerant subjects with cardiovascular Disease (TRANSCEND) trial conducted in ACEI-intolerant patients. In ONTARGET, telmisartan had a better tolerability profile with similar renoprotective properties compared with ramipril, suggesting a potential clinical benefit over ramipril. The recently completed Olmesartan Reducing Incidence of Endstage Renal Disease in Diabetic Nephropathy Trial (ORIENT) and Olmesartan and Calcium Antagonists Randomized (OSCAR) studies will further define the role of ARBs in cardioprotection and renoprotection for high-risk patients. PMID- 21633522 TI - Early follow-up results of arteriovenous fistulae created for hemodialysis. AB - BACKGROUND: The aim of this study was to evaluate the site, early results, and postoperative complications of arteriovenous fistula (AVF) creation procedures for hemodialysis in our clinic. METHODS: The hospital records of 384 patients who underwent 411 AVF creation procedures for hemodialysis by the same team at our clinic between February 2008 and January 2010 were included for retrospective analysis. All procedures were performed under local anesthesia with lidocaine. Vasospasm was treated by mechanical dilatation with a probe and topical papaverine. RESULTS: Of our 384 patients, 58.5% were male and 41.5% were female. Mean age was 46 (range 12-72) years. Of the 411 AVF procedures performed, 106 (25.8%) were created at the anatomical snuffbox, 264 (64.3%) were Brescia-Cimino procedures, and 41 (9.9%) were antecubital, brachiocephalic, or brachiobasilic procedures. Twenty-three patients (5.98%) were subjected to more than one surgical intervention due to early thrombosis or failure of AVF. Early patency was found in 94.0% of the AVF created. Twenty-three patients underwent more than one surgical intervention due to early AVF thrombosis or failure. Early AVF failure occurred more often in females (60.8%) than in males (39.2%). Complications were observed in a total of 11.4% patients. CONCLUSION: Mechanical dilatation of the artery and vein, before starting the anastomosis, as well as the use of vasodilatory agents, could decrease early thrombosis of the fistula, and this method has very high early patency. PMID- 21633523 TI - "It's time for your life": How should we remind patients to take medicines using short text messages? AB - The objective of this paper is to characterize effective patient care reminder strategies for people living with HIV/AIDS (PLWHA) to improve antiretroviral therapy (ART) adherence) using short message service (SMS) based on patient perspectives. We conducted a qualitative study with adult PLWHA in a community based clinic in Lima, Peru using focus groups. 26 HIV-positive individuals participated in four focus groups (20 men, 6 women). The participants expressed positive perceptions towards receiving reminders via SMS, but specified certain characteristics they wanted them to have (such as being simple and concise). It was also important that the messages maintained confidentiality and privacy by using coded words or phrases ("Remember, it is the time of your life") instead of "sensitive" words (HIV or antiretroviral). This study suggests that patients want healthcare SMS that appropriately notify them, deliver a careful crafted message, and assess the context in which they are received. PMID- 21633525 TI - Mallory-Denk Bodies in chronic hepatitis. AB - Mallory-Denk Bodies (MDB) are important as investigators, suggesting MDB as an indicator of the histologic severity of chronic hepatitis, causes of which include hepatitis C, primary biliary cirrhosis (PBC), and nonalcoholic fatty liver disease (NAFLD). Matteoni et al scored MDB in patients with NAFLD as none, rare and many, and reported that MDB plays a prominent role in this classification scheme in an earlier classification system. In this study, we evaluated 258 patients with chronic hepatitis due to metabolic, autoimmune and viral etiologies. Liver biopsy samples were evaluated with hematoxylin and eosin, periodic acid-Schiff-diastase, Gordon and Sweet's reticulin, Masson's trichrome, and iron stains. Both staging and grading were performed. Additionally, MDB were evaluated and discussed for each disease. We examined patients with nonalcoholic steatohepatitis (NASH; 50 patients), alcoholic hepatitis (10 patients), PBC (50 patients), Wilson disease (WD; 20 patients), hepatitis B (50 patients), hepatitis C (50 patients) and hepatocellular carcinoma (HCC; 30 patients). Frequency of MDB was as follows; NASH: 10 patients with mild in 60% and moderate in 40% and observed in every stage of the disease and frequently seen in zone 3. PBC: 11 patients with mild in 10%, moderate in 70%, and cirrhosis in 20%, and frequently seen in zone 1. WD: 16 patients with moderate and severe in 60% and cirrhosis in 40% and frequently seen in zone 1. Hep B: 3 patients with mild in 66% and severe in 34%. Hep C: 7 patients with mild in 40% and moderate in 60% and observed in every stage. HCC: 3 patients with hep B in 2 patients. We found that there is no relationship between MDB and any form of chronic hepatitis regarding histologic severity such as alcoholic steatohepatitis and NAFLD and variable zone distribution by etiology. PMID- 21633524 TI - Wound healing of intestinal epithelial cells. AB - The intestinal epithelial cells (IECs) form a selective permeability barrier separating luminal content from underlying tissues. Upon injury, the intestinal epithelium undergoes a wound healing process. Intestinal wound healing is dependent on the balance of three cellular events; restitution, proliferation, and differentiation of epithelial cells adjacent to the wounded area. Previous studies have shown that various regulatory peptides, including growth factors and cytokines, modulate intestinal epithelial wound healing. Recent studies have revealed that novel factors, which include toll-like receptors (TLRs), regulatory peptides, particular dietary factors, and some gastroprotective agents, also modulate intestinal epithelial wound repair. Among these factors, the activation of TLRs by commensal bacteria is suggested to play an essential role in the maintenance of gut homeostasis. Recent studies suggest that mutations and dysregulation of TLRs could be major contributing factors in the predisposition and perpetuation of inflammatory bowel disease. Additionally, studies have shown that specific signaling pathways are involved in IEC wound repair. In this review, we summarize the function of IECs, the process of intestinal epithelial wound healing, and the functions and mechanisms of the various factors that contribute to gut homeostasis and intestinal epithelial wound healing. PMID- 21633526 TI - Asymmetric dimethylarginine: a novel biomarker of gastric mucosal injury? AB - Nitric oxide (NO), a multifunctional endogenous gas molecule, is metabolized from L-arginine by enzymatic reaction in the presence of nitric oxide synthase. NO, an important gas signaling molecule, is a gastric mucosa protective factor that contributes significantly to maintain normal gastric mucosa integrity. NO increases gastric mucosa blood flow, regulates the secretion of mucus and bicarbonate, and inhibits the secretion of gastric juice. Asymmetric dimethylarginine (ADMA) has been identified as the major endogenous inhibitor of nitric oxide synthase. The function of ADMA is to decrease NO production via inhibiting nitric oxide synthase activity. Besides inhibiting NO synthesis, ADMA also directly induces oxidative stress and cell apoptosis, and participates in inflammation reaction. Its systemic accumulation was observed in conjunction with several cardiovascular and metabolic diseases. ADMA also mediates gastric ulcer injury induced by ethanol, stress, helicobacter pylori and indomethacin. The mechanism of ADMA directly producing adverse effect in gastric mucosa is incompletely understood. It is widely accepted that NO bioavailability decrease is the majority reason. Promotion of apoptosis and aggravation of inflammation may be other important mechanisms of ADMA-induced gastric injury. ADMA might be a novel clinical and experimental biomarker related to gastric mucosa disorder. Although therapeutic tool targeting to ADMA is available in multiple cardiovascular diseases, it is unknown in gastrointestinal disease. The strategy to inhibit ADMA is beneficial to gastric ulcer induced by ethanol in rats. Thus, ADMA might be a candidate of therapeutic target in gastric mucosa damage. PMID- 21633527 TI - Soluble ST2: a new and promising activity marker in ulcerative colitis. AB - AIM: To correlate circulating soluble ST2 (sST2) levels with the severity of ulcerative colitis (UC) and serum levels of pro-inflammatory cytokines, and to demonstrate the predictive power of sST2 levels for differentiation between active and inactive UC. METHODS: We recruited 153 patients: 82 with UC, 26 with Crohn's disease (CD) and 43 disease controls [non-inflammatory bowel disease (IBD)]. Subjects were excluded if they had diagnosis of asthma, autoimmune diseases or hypertension. The serum levels of sST2 and pro-inflammatory cytokines [pg/mL; median (25th-75th)] as well as clinical features, endoscopic and histological features, were subjected to analyses. The sST2 performance for discrimination between active and inactive UC, non-IBD and healthy controls (HC) was determined with regard to sensitivity and specificity, and Spearman's rank correlation coefficient (r). To validate the method, the area under the curve (AUC) of receiver-operator characteristic (ROC) was determined (AUC, 95% CI) and the total ST2 content of the colonic mucosa in UC patients was correlated with circulating levels of sST2. RESULTS: The serum sST2 value was significantly higher in patients with active [235.80 (90.65-367.90) pg/mL] rather than inactive UC [33.19 (20.04-65.32) pg/mL], based on clinical, endoscopic and histopathological characteristics, as well as compared with non-IBD and HC (P < 0.001). The median level of sST2 in CD patients was 54.17 (35.02-122.0) pg/mL, significantly higher than that of the HC group only (P < 0.01). The cutoff was set at 74.87 pg/mL to compare active with inactive UC in a multicenter cohort of patients. Values of sensitivity, specificity, and ability to correctly classify UC, according to activity, were 83.33%, 83.33% and 83.33%, respectively. The AUC of the ROC curve to assess the ability of this molecule to discriminate between active vs inactive UC was 0.92 (0.86-0.97, P < 0.0001). The serum levels of sST2 in patients with UC significantly correlated with endoscopic and histopathological scores (r = 0.76 and r = 0.67, P < 0.0001, respectively), and with the pro-inflammatory cytokine, tumor necrosis factor-alpha (r = 0.69 and r = 0.61, respectively, P < 0.0001). Interestingly, we found a direct correlation between total intestinal ST2 content and serum levels of sST2, adjusted to endoscopic activity score in patients with mild (r = 0.44, P = 0.004), moderate (r = 0.59, P = 0.002) and severe disease (r = 0.82, P = 0.002). Only patients with inactive UC showed no significant correlation (r = 0.45, P = 0.267). CONCLUSION: sST2 levels correlated with disease severity and inflammatory cytokines, are able to differentiate active from inactive UC and might have a role as a biomarker. PMID- 21633528 TI - Parallel decline of CD8+CD38+ lymphocytes and viremia in treated hepatitis B patients. AB - AIM: To assess the peripheral T lymphocyte subsets in chronic hepatitis B virus (HBV) infection, and their dynamics in response to adefovir dipivoxil monotherapy. METHODS: Proportions and absolute counts of peripheral natural killer cells, B cells, CD8+, CD4+, CD8+CD38+, CD8+CD28+ and CD4+CD28+ T cells were determined using three-color flow cytometry in chronic hepatitis B patients (n = 35), HBV carriers (n = 25) and healthy controls (n = 35). Adefovir dipivoxil was initiated in 17 chronic hepatitis B patients who were regularly followed for 72 wk, during which period the T cell subsets and serum viral load were measured at each follow-up point. RESULTS: The peripheral CD4+ T cell counts and CD8+ T cell counts decreased in chronic HBV infection. In chronic hepatitis B patients, proportions of CD8+CD38+ T cells were 62.0% +/- 14.7%, much higher than those of HBV carriers and healthy controls. In the 13 hepatitis B patients who were treated and responded to adefovir dipivoxil, proportions of CD8+CD38+ T cells decreased from 53.9% +/- 18.4% pre-therapy to 20.1% +/- 11.3% by week 72 (P < 0.001), concomitant with viral load decline (HBV DNA fell from 7.31 to 3 log copies/mL). CD8+ T cell counts also underwent an average increase of 218 cells/MUL by the end of 72-wk treatment. In those who failed the therapy, the CD8+CD38+ T cell population had more fluctuations. CONCLUSION: CD8+ T cells abnormally activated in chronic HBV infection can be partially reversed by antiviral therapy. HBV-associated immune activation may be a crucial part of the pathogenesis and a promising target of treatment. PMID- 21633530 TI - Value of transient elastography for the prediction of variceal bleeding. AB - AIM: To determine if liver stiffness (LS) measurements by means of transient elastography (TE) correlate with the presence of significant esophageal varices (EV) and if they can predict the occurrence of variceal bleeding. METHODS: We studied 1000 cases of liver cirrhosis divided into 2 groups: patients without EV or with grade 1 varices (647 cases) and patients with significant varices (grade 2 and 3 EV) (353 cases). We divided the group of 540 cases with EV into another 2 subgroups: without variceal hemorrhage (375 patients) and patients with a history of variceal bleeding (165 cases). We compared the LS values between the groups using the unpaired t-test and we established cut-off LS values for the presence of significant EV and for the risk of bleeding by using the ROC curve. RESULTS: The mean LS values in the 647 patients without or with grade 1 EV was statistically significantly lower than in the 353 patients with significant EV (26.29 +/- 0.60 kPa vs 45.21 +/- 1.07 kPa, P < 0.0001). Using the ROC curve we established a cut-off value of 31 kPa for the presence of EV, with 83% sensitivity (95% CI: 79.73%-85.93%) and 62% specificity (95% CI: 57.15%-66.81%), with 76.2% positive predictive value (PPV) (95% CI: 72.72%-79.43%) and 71.3% negative predictive value (NPV) (95% CI: 66.37%-76.05%) (AUROC 0.7807, P < 0.0001). The mean LS values in the group with a history of variceal bleeding (165 patients) was statistically significantly higher than in the group with no bleeding history (375 patients): 51.92 +/- 1.56 kPa vs 35.20 +/- 0.91 kPa, P < 0.0001). For a cut-off value of 50.7 kPa, LS had 53.33% sensitivity (95% CI: 45.42%-61.13%) and 82.67% specificity (95% CI: 78.45%-86.36%), with 82.71% PPV (95% CI: 78.5%-86.4%) and 53.66% NPV (95% CI: 45.72%-61.47%) (AUROC 0.7300, P < 0.0001) for the prediction of esophageal bleeding. CONCLUSION: LS measurement by means of TE is a reliable noninvasive method for the detection of EV and for the prediction of variceal bleeding. PMID- 21633529 TI - Hepatotropic growth factors protect hepatocytes during inflammation by upregulation of antioxidative systems. AB - AIM: To investigate effects of hepatotropic growth factors on radical production in rat hepatocytes during sepsis. METHODS: Rat hepatocytes, isolated by collagenase perfusion, were incubated with a lipopolysaccharide (LPS)-containing cytokine mixture of interleukin-1beta, tumor necrosis factor-alpha and interferon gamma to simulate sepsis and either co-incubated or pre-incubated with hepatotropic growth factors, e.g. hepatocyte growth factor, epidermal growth factor and/or transforming growth factor-alpha. Cells were analyzed for glutathione levels. Culture supernatants were assayed for production of reactive oxygen intermediates (ROIs) as well as NO(2) (-), NO(3) (-) and S-nitrosothiols. To determine cellular damage, release of aspartate aminotransferase (AST) into the culture medium was analyzed. Activation of nuclear factor (NF)-kappaB was measured by electrophoretic mobility shift assay. RESULTS: Rat hepatocytes treated with the LPS-containing cytokine mixture showed a significant increase in ROI and nitrogen oxide intermediate formation. AST leakage was not significantly increased in cells treated with the LPS-containing cytokine mixture, independent of growth-factor co-stimulation. However, pretreatment with growth factors significantly reduced AST leakage and ROI formation while increasing cellular glutathione. Application of growth factors did not result in increased NF-kappaB activation. Pretreatment with growth factors further increased formation of NO(2) (-), NO(3) (-) and S-nitrosothiols in hepatocytes stimulated with LPS-containing cytokine mixture. Thus, we propose that, together with an increase in glutathione increased NO(2) (-), NO(3) (-) formation might shift their metabolism towards non toxic products. CONCLUSION: Our data suggest that hepatotropic growth factors positively influence sepsis-induced hepatocellular injury by reducing cytotoxic ROI formation via induction of the cellular protective antioxidative systems. PMID- 21633531 TI - Increased intestinal permeability in inflammatory bowel diseases assessed by iohexol test. AB - AIM: To study intestinal permeability (IP) and its relationship to the disease activity in patients with inflammatory bowel diseases (IBD) - Crohn's disease (CD) and ulcerative colitis (UC). METHODS: Fifty-eight patients with active IBD (32 with CD and 26 with UC) and 25 healthy controls consented to participate in the study. The clinical activity of CD was estimated using the Crohn's Disease Activity Index (CDAI), and the endoscopic activity of UC using the Mayo scoring system. IP was assessed by the rise in levels of iohexol, which was administered orally (25 mL, 350 mg/mL) 2 h after breakfast. Three and six hours later serum (SIC mg/L) and urine (UIC g/mol) iohexol concentrations were determined by a validated HPLC-UV technique. RESULTS: In the CD group, SIC values at 3 h (2.95 +/ 2.11 mg/L) and at 6 h after ingestion (2.63 +/- 2.18 mg/L) were significantly higher compared to those of healthy subjects (1.25 +/- 1.40 mg/L and 1.11 +/- 1.10 mg/L, respectively, P < 0.05). UIC (g/mol) values were also higher in patients, but the differences were significant only for UIC at 6 h. Significant positive correlation (P < 0.05) was found between the CDAI and IP, assessed by SIC at 3 h (r = 0.60) and 6 h (r = 0.74) after the ingestion. In comparison to controls, SIC and UIC of UC patients were higher in the two studied periods, but the differences were significant at 6 h only. Significantly higher values of SIC (P < 0.05) were found in patients with severe endoscopic activity of UC compared to those of patients with mild and moderate activity (3.68 +/- 3.18 vs 0.92 +/- 0.69 mg/L). CONCLUSION: Serum levels of iohexol at 3 h and 6 h after its ingestion reflect increased IP, which is related to the disease activity in patients with IBD. PMID- 21633532 TI - Pre-operative factors that can predict neoplastic polypoid lesions of the gallbladder. AB - AIM: To investigate the preoperative factors that can predict neoplastic polypoid lesions of the gallbladder (PLGs) as well as malignant PLGs. METHODS: A retrospective analysis was conducted on the 210 consecutively enrolled patients who underwent cholecystectomy due to a PLG larger than 10 mm, as was determined by preoperative trans-abdominal ultrasonography or endoscopic ultrasonography. We analyzed the medical, laboratory, radiologic data and the pathologic results. RESULTS: In 210 cases, 146 had non-neoplastic polyps (69.5%) and 64 cases were neoplastic polyps (30.5%). An older age (>= 65 years), the presence of diabetes mellitus (DM) and the size of polyp (>= 15 mm) were revealed to be independent predictive variables for neoplastic polyps with odd ratios (OR) of 2.27 (P = 0.044), 2.64 (P = 0.021) and 4.94 (P < 0.01), respectively. Among the neoplastic PLGs, an older age (>= 65 years), the presence of DM and polyp size (>= 15 mm) were associated with malignancy with ORs of 4.97 (P = 0.005), 6.13 (P = 0.001) and 20.55 (P < 0.001), respectively. CONCLUSION: Among patients with PLGs larger than 10 mm in size, higher risk groups such as elderly patients more than 65 years old, those with DM or a large polyp size (>= 15 mm) should be managed by cholecystectomy. PMID- 21633533 TI - Topical application of glycyrrhizin preparation ameliorates experimentally induced colitis in rats. AB - AIM: To examine the efficacy of glycyrrhizin preparation (GL-p) in the treatment of a rat model of ulcerative colitis (UC). METHODS: Experimental colitis was induced by oral administration of dextran sodium sulfate. Rats with colitis were intrarectally administered GL-p or saline. The extent of colitis was evaluated based on body weight gain, colon wet weight, and macroscopic damage score. The expression levels of pro-inflammatory cytokines and chemokines in the inflamed mucosa were measured by cytokine antibody array analysis. The effect of GL-p on myeloperoxidase (MPO) activity in the inflamed mucosa and purified enzyme was assayed. RESULTS: GL-p treatment significantly ameliorated the extent of colitis compared to sham treatment with saline. Cytokine antibody array analysis showed that GL-p treatment significantly decreased the expression levels of pro inflammatory cytokines and chemokines, including interleukin (IL)-1beta, IL-6, tumor necrosis factor-alpha, cytokine-induced neutrophil chemoattractant-2, and monocyte chemoattractant protein-1 in the inflamed mucosa. Furthermore, GL-p inhibited the oxidative activity of mucosal and purified MPO. CONCLUSION: GL-p enema has a therapeutic effect on experimental colitis in rats and may be useful in the treatment of UC. PMID- 21633534 TI - Hyperbaric oxygenation promotes regeneration of biliary cells and improves cholestasis in rats. AB - AIM: To investigate the effects of hyperbaric oxygenation (HBO) on regeneration of the biliary ductal system and postoperative cholestasis in hepatectomized rats. METHODS: HBO was performed in Wistar rats daily starting 12 h after a 70% partial hepatectomy. Regenerated liver weight, serum parameters and the proliferating cell nuclear antigen labeling index of hepatocytes and biliary ductal cells were measured. Hepatocyte growth factor (HGF), c-Met and transforming growth factor (TGF) beta-1 mRNA expression levels were analyzed by quantitative reverse transcription polymerase chain reaction. RESULTS: HBO improved the postoperative serum levels of total bile acid but not transaminase levels. HBO promoted hepatocyte and biliary ductal cell proliferation. The hematoxylin and eosin-stained specimens revealed fewer ballooned hepatocytes and higher cell densities in the HBO group compared to the control group. HBO suppressed c-Met mRNA levels at 15 h but did not modulate HGF or TGF beta-1 mRNA expression levels. CONCLUSION: HBO promoted regeneration of biliary ductal cells and improved postoperative cholestasis after a partial hepatectomy. PMID- 21633535 TI - Limited water infusion decreases pain during minimally sedated colonoscopy. AB - AIM: To investigate a limited water infusion method in colonoscopy. METHODS: Consecutive patients undergoing minimally sedated colonoscopy were randomized to receive air insufflation (n = 89) or water infusion limited to the rectum, sigmoid colon and descending colon (n = 90). Completion rates, cecal intubation times, procedure times, need for abdominal compression, turning of patients and levels of discomfort were evaluated. RESULTS: Completion rates, total procedure times, need for abdominal compression, and turning of patients were similar between groups. Less pain was experienced in the water group than in the air group (2.5 +/- 2.5 vs 3.4 +/- 2.8, mean +/- SD, P = 0.021). The cecal intubation time was significantly longer in the water group than in the air group (6.4 +/- 3.1 min vs 4.5 +/- 2.4 min, P < 0.001). More water was infused in the water group (322 +/- 80.9 mL vs 26.2 +/- 39.4 mL, P < 0.001). CONCLUSION: Limited airless water infusion in the distal colon reduces patients' pain during colonoscopy. PMID- 21633536 TI - Mechanism and dose-effect of Ginkgolide B on severe acute pancreatitis of rats. AB - AIM: To determine the optimal dosage and mechanism of Ginkgolide B (BN52021) on severe acute pancreatitis (SAP) of rats. METHODS: Seventy male Wistar rats were randomly divided into seven groups (10 for each group). Sham-operation group (SO), SAP model group (SAP), dimethyl sulfoxide (DMSO) contrast group (DMSO), and groups treated with 2.5 mg/kg BN52021 (BN1), 5 mg/kg BN52021 (BN2), 10 mg/kg BN52021 (BN3), and 20 MUg/kg Sandostatin (SS). The SAP model was established in Wistar rats by injecting 5% sodium taurocholate retrogradely into the common bilio-pancreatic duct. The rats of SO, DMSO and BN52021 were injected with 0.9% NaCl, 0.5% DMSO and BN52021 through femoral vein 15 min after the operation. The SS group was injected with Sandostatin subcutaneously. All rats were anaesthetized at 6 h after operation, and venous blood was collected to determine the levels of serum amylase and phospholipase A2 (PLA2), and pancreas tissue was harvested and stained. RESULTS: There was no significant difference between the SAP and DMSO groups in serum amylase level, PLA2, ascites and pathologic score, but significant difference was found in SAP/DMSO groups compared with those in SO group (P < 0.05) and the levels of serum amylase, PLA2, ascites, and pathologic score were lower in the BN1, BN2, BN3 and SS groups than in the SAP and DMSO groups (P < 0.05). However, among BN1, BN2, BN3 and SS groups, BN2 had the best effect in decreasing the levels of serum amylase and PLA2 (P < 0.05). Expression of platelet activating factor (PAF) receptor (PAFR) mRNA and protein showed no significant difference between the SAP and DMSO groups, or among BN1, BN2, BN3 and SS groups, but there was remarkable difference between SAP/DMSO group and SO group (P < 0.05), and expression of PAFR mRNA and protein was higher in the BN1, BN2, BN3 and SS groups than in the SAP and DMSO groups (P < 0.05). PAFR expression was observed in the nucleus and cytoplasm of pancreatic islet cells in Wistar rats by immunohistochemistry. CONCLUSION: By iv injection, 5 mg/kg of BN52021 is the optimal dosage for SAP rats. BN52021 may inhibit the interaction/binding of PAF with PAFR. PMID- 21633537 TI - Specific HLA-DQB1 alleles associated with risk for development of hepatocellular carcinoma: a meta-analysis. AB - AIM: To evaluate the association of human leukocyte antigen (HLA)-DQB1 alleles with hepatocellular carcinoma (HCC) through meta-analysis of published data. METHODS: Case-control studies on HLA-DQB1 allele association with HCC published up to January 2010 were included in the analyses. The odds ratios (ORs) of HLA DQB1 allele distributions in HCC patients were analyzed and compared with healthy controls. The meta-analysis software REVMAN 5.0 was applied for investigating heterogeneity among individual studies and for summarizing all the studies. A meta-analysis was performed using fixed-effect or random-effect methods, depending on the absence or presence of significant heterogeneity. Seven case control studies containing 398 cases and 594 controls were included in the final analysis. RESULTS: Among the five family alleles, two (DQB1*02 and DQB1*03) were found to be significantly associated with the risk of HCC. The combined OR for the association of DQB1*02 and DQB1*03 allele with the risk for HCC was 1.78 (95% CI: 1.05-3.03, P = 0.03) and 0.65 (95% CI: 0.48-0.89, P = 0.007), respectively. Among the 13 specific alleles, two (DQB1*0502 and DQB1*0602) were significantly associated with risk of HCC. The combined OR for the association of DQB1*0502 and DQB1*0602 allele with the risk for HCC was 1.82 (95% CI: 1.14-2.92, P = 0.01) and 0.58 (95% CI: 0.36-0.95, P = 0.03), respectively. No significant association was established for other HLA-DQB1 family alleles and specific alleles. CONCLUSION: Our results support the hypothesis that specific HLA-DQB1 allele families and alleles might influence the susceptibility or resistance to HCC, although it needs further investigations. PMID- 21633538 TI - Neoadjuvant sorafenib combined with gemcitabine plus oxaliplatin in advanced hepatocellular carcinoma. AB - This paper reports the first case of a patient with hepatocellular carcinoma with lymph node metastasis treated by sorafenib combined with gemcitabine plus oxaliplatin, with a partial response and normalization of alpha fetoprotein, which allowed curative surgery. The potential synergy between these three drugs needs to be confirmed, and is currently being investigated in a randomized phase II trial. PMID- 21633539 TI - Significance of serum procalcitonin in sepsis. AB - CONTEXT: Rapid treatment of sepsis is of crucial importance for survival of patients. Specific and rapid markers of bacterial infection have been sought for early diagnosis of sepsis. One such measurement, Procalcitonin (PCT), has recently become of interest as a possible marker of the systemic inflammatory response to infection. AIMS: This study was done to find out the common sources of sepsis and to evaluate the diagnostic value of PCT, its predictive value and its relation with Sepsis-related Organ Failure Assessment (SOFA) scores and mortality in various stages of sepsis. SETTINGS AND DESIGN: The prospective study was conducted at our tertiary care center from October 2006 to December 2008. A total of 100 patients were included in the study. The study sample included all patients aged above 18 years presenting consecutively to our center during the study period with acute sepsis. They were divided into three groups: sepsis, severe sepsis and septic shockbased on standardized criteria. MATERIALS AND METHODS: PCT and various other relevant factors were measured in all study subjects. These parameters were compared among the three study groups. The statistical analyses were done using Student "t" test and two-way analysis of variance (ANOVA). RESULTS: Respiratory tract infection was the most common source of sepsis. PCT proved to be an excellent indicator of sepsis with sensitivity of 94%. There was a significant association between serum PCT and SOFA scores (P < 0.05). Serum PCT levels did not predict mortality in the present study. CONCLUSIONS: PCT is among the most promising sepsis markers, capable of complementing clinical signs and routine lab parameters suggestive of severe infection. PMID- 21633540 TI - Efficacy of a multimodal intervention strategy in improving hand hygiene compliance in a tertiary level intensive care unit. AB - CONTEXT: The role of hand hygiene in preventing health care associated infections (HCAIs) has been clearly established. However, compliance rates remain poor among health care personnel. AIMS: a) To investigate the health care workers' hand hygiene compliance rates in the intensive care unit (ICU), b) to assess reasons for non-compliance and c) to study the efficacy of a multimodal intervention strategy at improving compliance. SETTINGS: A mixed medical-surgical ICU of a tertiary level hospital. DESIGN: A before-after prospective, observational, intervention study. MATERIALS AND METHODS: All health care personnel who came in contact with patients in the ICU were observed for their hand hygiene compliance before and after a multimodal intervention strategy (education, posters, verbal reminders and easy availability of products). A self-report questionnaire was also circulated to assess perceptions regarding compliance. Statistical analysis was done using chi(2) test or Fisher exact test (Epi info software). RESULTS: Hand hygiene compliance among medical personnel working in the ICU was 26% and the most common reason cited for non-compliance was lack of time (37%). The overall compliance improved significantly following the intervention to 57.36% (P<0.000). All health care worker groups showed significant improvements: staff nurses (21.48-61.59%, P<0.0000), nursing students (9.86-33.33%, P<0.0000), resident trainees (21.62-60.71%, P<0.0000), visiting consultants (22-57.14%, P=0.0001), physiotherapists (70-75.95%, P=0.413) and paramedical staff (10.71 55.45%, P< 0.0000). CONCLUSIONS: Hand hygiene compliance among health care workers in the ICU is poor; however, intervention strategies, such as the one used, can be useful in improving the compliance rates significantly. PMID- 21633541 TI - Comparison of bronchoscopic and non-bronchoscopic techniques for diagnosis of ventilator associated pneumonia. AB - BACKGROUND: The diagnosis of ventilator associated pneumonia (VAP) remains a challenge because the clinical signs and symptoms lack both sensitivity and specificity and the selection of microbiologic diagnostic procedure is still a matter of debate. AIMS AND OBJECTIVE: To study the role of various bronchoscopic and non-bronchoscopic diagnostic techniques for diagnosis of VAP. SETTINGS AND DESIGN: This prospective comparative study was conducted in a medical ICU of a tertiary care center. MATERIALS AND METHODS: Twenty-five patients, clinically diagnosed with VAP, were evaluated by bronchoscopic and non-bronchoscopic procedures for diagnosis. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of various bronchoscopic and non bronchoscopic techniques were calculated, taking clinical pulmonary infection score (CPIS) of >=6 as reference standard. RESULTS: Our study has shown that for the diagnosis of VAP, bronchoscopic brush had a sensitivity, specificity, PPV and NPV of 94.9% [confidence interval (CI): 70.6-99.7], 57.1% (CI: 13.4-86.1), 85% (CI: 61.1-96) and 80% (CI: 21.9-98.7), respectively. Bronchoscopic bronchoalveolar lavage (BAL) had a sensitivity, specificity, PPV and NPV of 77.8% (CI: 51.9-92.6), 71.8% (CI: 24.1-94), 87.3% (CI: 60.4-97.8) and 55.5% (CI: 17.4 82.6), respectively. Sensitivity, specificity, PPV and NPV for non-bronchoscopic BAL (NBAL) were 83.3% (CI: 57.7-95.6), 71.43% (CI: 24.1-94), 88.2% (CI: 62.3 97.4) and 62.5% (CI: 20.2-88.2), respectively. Endotracheal aspirate (ETA) yield was only 52% and showed poor concordance with BAL (kappa-0.351; P-0.064) and NBAL (k-0.272; P-0.161). There was a good microbiologic concordance among different bronchoscopic and non-bronchoscopic distal airway sampling techniques. CONCLUSION: NBAL is an inexpensive, easy, and useful technique for microbiologic diagnosis of VAP. Our findings, if verified, might simplify the approach for the diagnosis of VAP. PMID- 21633542 TI - Electrolytes assessed by point-of-care testing - Are the values comparable with results obtained from the central laboratory? AB - BACKGROUND AND AIMS: When dealing with very sick patients, the speed and accuracy of tests to detect metabolic derangements is very important. We evaluated if there was agreement between whole blood electrolytes measured by a point-of-care device and serum electrolytes measured using indirect ion-selective electrodes. MATERIALS AND METHODS: In this prospective study, electrolytes were analyzed in 44 paired samples drawn from critically ill patients. Whole blood electrolytes were analyzed using a point-of-care blood gas analyzer and serum electrolytes were analyzed in the central laboratory on samples transported through a rapid transit pneumatic system. Agreement was summarized by the mean difference with 95% limits of agreement (LOA) and Lin's concordance correlation (p(c)). RESULTS: There was a significant difference in the mean (+/-standard deviation) sodium value between whole blood and serum samples (135.8 +/- 5.7 mmol/L vs. 139.9 +/- 5.4 mmol/L, P < 0.001), with the agreement being modest (p(c) = 0.71; mean difference -4.0; 95% LOA -8.78 to 0.65). Although the agreement between whole blood and serum potassium was good (p(c) = 0.96), and the average difference small (-0.3; 95% LOA -0.72 to 0.13), individual differences were clinically significant, particularly at lower potassium values. For potassium values <3.0 mmol/L, the concordance was low (p(c) = 0.53) and the LOA was wide (1.0 to 0.13). The concordance for potassium was good (p(c) = 0.96) for values >=3.0 (mean difference -0.2; 95% LOA -0.48 to 0.06). CONCLUSIONS: Clinicians should be aware of the difference between whole blood and serum electrolytes, particularly when urgent samples are tested at point of care and routine follow-up electrolytes are sent to the central laboratory. A correction factor needs to be determined at each center. PMID- 21633543 TI - An evaluation of the Charlson co-morbidity score for predicting sepsis after elective major surgery. AB - BACKGROUND AND AIMS: Severe sepsis is a significant cause of morbidity and mortality following major surgery. The Charlson co-morbidity score (CCS) has been shown to be associated with severe sepsis following major surgery for cancer. This prospective observational study investigated the effect of patient factors (CCS, gender, age and malignancy) and intraoperative factors (duration of surgery and allogeneic blood transfusion) on the incidence of sepsis after elective major surgery, and the impact of patient co-morbidities on length of stay in critical care. MATERIALS AND METHODS: We prospectively identified a cohort of 101 patients undergoing elective major surgery in a university teaching hospital. The CCS was calculated before surgery, and the incidence of sepsis was documented following surgery. We investigated whether age, malignancy, intraoperative allogeneic blood transfusion, length of surgery or gender were associated with sepsis following surgery. RESULTS: Twenty-seven (27%) patients developed sepsis. Using multivariate logistic regression, the duration of surgery was associated with the development of sepsis after surgery (P = 0.054, odds ratio 1.2). The CCS was not associated with sepsis in this population of cancer and non-cancer patients undergoing elective major surgery, but was associated with longer length of stay in the intensive care unit (P = 0.016). CONCLUSIONS: Duration of surgery, but not patient co-morbidity as assessed by the CCS, may predict the postoperative incidence of sepsis. CCS could be used as a guide to predict consumption of critical care resources by elective surgical patients. A higher CCS was associated with a longer ICU stay. Resources, such as postoperative goal directed therapy, may be useful in reducing length of stay, hospital costs and risks of infective complications in this subgroup of patients with higher CCS. PMID- 21633544 TI - Anaphylactic reaction to intravenous diclofenac. AB - Diclofenac sodium is a non-steroidal anti-inflammatory drug widely used as an opioid sparing agent for postoperative analgesia. Anaphylaxis due to intravenous diclofenac sodium is very rare. We report a case of anaphylactic reaction to IV diclofenac sodium, occurring postoperatively in a 25-year-old primigravida, the clinical features of which mimicked pulmonary embolism. The rarity, clinical importance and the diagnostic dilemma associated prompted us to report this case. PMID- 21633545 TI - Hemodialysis through persistent left superior vena cava. AB - We report a case of end stage renal disease patient who displayed a persistent left superior vena cava (PLSVC) after placement of hemodialysis (HD) catheter through left internal jugular vein, as revealed by routine post-procedure X-ray chest. The diagnosis of PLSVC was confirmed by arterial blood gas, two dimensional echocardiography, computed tomography thorax and angiographic examination. This anomaly is rather rare; few studies on safety of PLSVC for HD have been reported. The catheter was uneventfully used for HD for 2 months with careful continuous monitoring and removed after arteriovenous fistula was successfully cannulated. Physicians who place HD catheters in the left jugular/subclavian vein should be aware of the existence of PLSVC. PMID- 21633546 TI - Apical ballooning syndrome after attempted suicidal hanging. AB - We report a case of "Apical Ballooning Syndrome" following attempted suicidal hanging. Our patient developed retrosternal chest pain and ischemic changes on electrocardiography (ECG), a day after the suicidal attempt. She underwent an angiogram considering the possibility of acute coronary syndrome. However, her coronary arteries were normal; the left ventricle showed the typical ballooning pattern characterized by hypokinesia of the distal septum and apex. On follow-up a week later, she remained asymptomatic; her ECG changes had reversed and the left ventricular contractility was normal on echocardiography. PMID- 21633547 TI - Nonchylous idiopathic pleural effusion in the newborn. AB - Congenital isolated pleural effusion is a rare cause of respiratory distress in neonates. It is usually chylous. Herein, we report a rare case of nonchylous congenital idiopathic pleural effusion. PMID- 21633548 TI - Post-traumatic skin and soft tissue infection due to Aeromonas hydrophila. AB - We report a case of posttraumatic skin and soft tissue infection in a patient who sustained laceration after being hit by a water tanker. Aeromonas hydrophila was isolated from pus and was identified to the species level by Vitek 2 and a battery of biochemical tests. The patient responded to thorough drainage, debridement of wound and 2 weeks of intravenous antibiotics. The patient was taken up for split skin grafting of the raw area. She was discharged with satisfactory graft uptake after 1 week without any further antibiotics advice. Follow-up after 3 weeks was satisfactory with healthy cover on the raw area and normal weight bearing on the left leg. PMID- 21633549 TI - Contralateral hyperinflation: Computed tomography demonstration of an unusual complication of unrecognized endobronchial intubation. AB - Endobronchial intubation (EBI) is an important complication of endotracheal intubation. In a case of unrecognized EBI, usually, the intubated lung gets hyperinflated while the contralateral lung collapses. We report a case of unrecognized right main stem EBI with ipsilateral normal aeration and contralateral hyperinflation detected during computed tomography scan of the chest for trauma work up in a case of severe head injury. PMID- 21633550 TI - Drug interaction leading to prolonged sedation in a postoperative high risk coronary bypass surgery patient. AB - Use of midazolam infusion in mechanically ventilated patient is an established practice in critical care. In our case, the use of erythromycin as a prokinetic agent for better tolerance of enteral feeding and paralytic ileus led to an interaction between midazolam and erythromycin, which resulted in prolonged and deeply sedated patient. In a critically ill patient, there is always a possibility of multiple drug interactions. It is important to understand them and they should be considered before starting new medication. PMID- 21633551 TI - Revisiting the eye opening response of the Glasgow Coma Scale. AB - The Glasgow Coma Scale (GCS), introduced by Teasdale and Jenneth in 1974, has received tremendous acclaim from clinicians and has been extensively used in clinical practice for the evaluation of the level of consciousness. The author notes that some traumatic brain injury patients close eyes in response to painful stimuli as opposed to the eye opening response to pain of the GCS. A revision of the eye opening response subsection of the GCS is suggested. PMID- 21633552 TI - Novel H1N1 influenza infection in intensive care unit. PMID- 21633553 TI - Authors' reply. PMID- 21633554 TI - Right subclavian artery cannulation: Is chest roentgenogram sufficient to diagnose the complication? PMID- 21633555 TI - Author's reply. PMID- 21633556 TI - Critically ill patients with 2009 H1N1 infection in an Indian intensive care unit. PMID- 21633557 TI - Encephalopathy as a presenting feature of ascariasis in a child. PMID- 21633558 TI - What's new in Emergencies, Trauma and Shock? Anesthesia, surgery and postoperative cognition. PMID- 21633559 TI - JETS policy on plagiarism and academic dishonesty. PMID- 21633560 TI - A clinico-epidemiologic study of 892 patients with burn injuries at a tertiary care hospital in Punjab, India. AB - AIM: To analyze the causes, demographic and socio-cultural aspects, and the magnitude of burn injuries prospectively and to evaluate the outcome of treatment of patients admitted to burns ICU of tertiary care hospital. MATERIALS AND METHODS: A total of 892 burn patients admitted over a period of 6 years from January 2002 to December 2007 at tertiary care hospital in Punjab, India, were analyzed. RESULTS: 54% patients were males. A majority of the patients, 704(79%), were in the age group of 15-45 years. Six hundred and thirty-four patients (72%) sustained flame burns, while 17% and 7% sustained electrical and scald burns, respectively. A total of 470(53%) patients sustained major two to three degree flame burns involving more than 45% of total body surface area (TBSA). The mortality rate was 40%, i.e. 357 patients died of burns and its related problems, in our study. Six hundred and thirty-nine patients (72%) sustained burns in closed space of which 331 patients (52%) sustained burns in kitchen. Seven hundred and seventy-nine patients sustained accidental burns. Burn victims were mainly Hindus and Sikhs. The mean hospital stay varied depending upon the percentage of burns. On an average, a patient with > 45% TBSA burns received 15 whole blood transfusions. Split skin grafting was done in 416 patients. Most common complication encountered during their hospital stay was wound infection which was seen in 671 patients, followed by ARDS in 221 patients. The most common organisms causing wound infection were Pseudomonas and Acinetobacter. CONCLUSION: Developing country like India need an aggressive public education program so that people become more literate about various etiological factors causing burns and means of preventing them. Also needed are burn care hospitals which are easily accessible and affordable. PMID- 21633561 TI - Pediatric trauma mortality by type of designated hospital in a mature inclusive trauma system. AB - BACKGROUND: Previous studies have shown divergent results regarding the survival of injured children treated at pediatric trauma centers (PTC) and adult trauma centers (ATC). AIMS: (1) To document, in a regionalized inclusive trauma system, at which level of trauma centers were the injured children treated and (2) to compare the in-hospital mortality over five levels of trauma care, ranging from pediatric level I trauma centers (PTC) to designated local trauma hospitals (level IV) for the whole study sample and for subgroups of severely injured children and head trauma. MATERIALS AND METHODS: A retrospective analysis included data on 11,053 injured children (age <=16 years) treated between April 1998 and March 2005 in 58 designated trauma hospitals in the province of Quebec, Canada. Multiple imputation was used to handle missing physiological data and multivariate logistic regression was used to compare mortality over levels of care. RESULTS: PTC treated 52.2% of the children. Children treated at PTC were more often transferred from another hospital (73%) and were more severely injured. ATC level I, II, III and IV centers treated, respectively, 3.0%, 16.2%, 24.3% and 4.3% of children. Compared with children treated at a PTC, the risk of mortality was higher for children treated at each other ATC, i.e. level I (adjusted odds ratio [OR] = 3.1; 95% confidence interval [CI]: 1.3-7.5), level II (OR = 2.5; 95% CI: 1.3-5.0), level III (OR = 5.2; 95% CI: 2.1-13.1) and level IV (OR = 9.9; 95% CI: 2.4-41.3). Similar findings were observed among the subsamples of children who were more severely injured (Injury Severity Score >15) and who sustained head injuries. CONCLUSIONS: In our trauma system, PTC cared for more than half of the injured children and patients treated there have better survival than those treated at all other levels of ATC. PMID- 21633562 TI - Vascular injuries after bear attacks: Incidence, surgical challenges and outcome. AB - BACKGROUND: Bear mauling is rarely reported in medical literature due to its rare occurrence. Present study was undertaken to describe the pattern and management of bear maul vascular injuries in Kashmir. PATIENTS AND METHODS: Study of patients with bear maul vascular injury from 1(st) Jan 2004 to 31(st) Dec. 2008. Fifteen patients with bear maul vascular injury were studied. All patients of bear maul without vascular injury were excluded from the study. RESULTS: Most of the patients were treated by reverse saphenous vein graft or end to end anastomosis. Most common complication was wound infection (20%) followed by graft occlusion (13.33%). There was no operative death. CONCLUSION: Bear attacks are very common in Kashmir. Vascular injury due to bear maul needs prompt resuscitation and revascularization. Results are very good provided timely intervention for revascularization is done. PMID- 21633563 TI - Impact of general versus epidural anesthesia on early post-operative cognitive dysfunction following hip and knee surgery. AB - BACKGROUND: Post-operative cognitive dysfunction is the subtle cerebral complication temporally seen following surgery. The aim of this study was to compare the influence of either general anesthesia (GA) or epidural anesthesia (EA) on the early post-operative neurocognitive outcome in elderly (>59 years) subjects undergoing hip and knee surgery. METHODS: A total of 60 patients were recruited in a prospective, randomized, parallel-group study, comparable by age and sex. They were enrolled and randomized to receive either EA (n = 30) or GA (n = 30). All of them were screened using the Mini Mental State Examination (MMSE), with components of the Kolkata Cognitive Screening Battery. The operated patients were re-evaluated 1 week after surgery using the same scale. The data collected were analyzed to assess statistical significance. RESULTS: We observed no statistical difference in cognitive behavior in either group pre-operatively, which were comparable with respect to age, sex and type of surgery. Grossly, a significant difference was seen between the two groups with respect to the perioperative changes in verbal fluency for categories and MMSE scores. However, these differences were not significant after the application of the Bonferroni correction for multiple analyses, except the significant differences observed only in the MMSE scores. CONCLUSIONS: We observed a difference in cognitive outcome with GA compared with EA. Certain aspects of the cognition were affected to a greater extent in this group of patients undergoing hip and knee surgery. PMID- 21633564 TI - Isolated traumatic head injury in children: Analysis of 276 observations. AB - BACKGROUND: To determine predictive factors of mortality among children after isolated traumatic brain injury. MATERIALS AND METHODS: In this retrospective study, we included all consecutive children with isolated traumatic brain injury admitted to the 22-bed intensive care unit (ICU) of Habib Bourguiba University Hospital (Sfax, Tunisia). Basic demographic, clinical, biochemical, and radiological data were recorded on admission and during ICU stay. RESULTS: There were 276 patients with 196 boys (71%) and 80 girls, with a mean age of 6.7 +/- 3.8 years. The main cause of trauma was road traffic accident (58.3%). Mean Glasgow Coma Scale score was 8 +/- 2, Mean Injury Severity Score (ISS) was 23.3 +/- 5.9, Mean Pediatric Trauma Score (PTS) was 4.8 +/- 2.3, and Mean Pediatric Risk of Mortality (PRISM) was 10.8 +/- 8. A total of 259 children required mechanical ventilation. Forty-eight children (17.4%) died. Multivariate analysis showed that factors associated with a poor prognosis were PRISM > 24 (OR: 10.98), neurovegetative disorder (OR: 7.1), meningeal hemorrhage (OR: 2.74), and lesion type VI according to Marshall tomographic grading (OR: 13.26). CONCLUSION: In Tunisia, head injury is a frequent cause of hospital admission and is most often due to road traffic injuries. Short-term prognosis is influenced by demographic, clinical, radiological, and biochemical factors. The need to put preventive measures in place is underscored. PMID- 21633565 TI - Initial systolic blood pressure and ongoing internal bleeding following torso trauma. AB - OBJECTIVE: Recent studies have suggested that an initial systolic blood pressure (SBP) in the range of 90-110 mmHg in a trauma patient may be indicative of hypoperfusion and is associated with poor patient outcome. However, the use of initial SBP as a surrogate for predicting internal bleeding is yet to be validated. The purpose of this study was to assess the presenting SBPs in patients with torso trauma and evidence of ongoing internal hemorrhage. SETTING AND DESIGN: This was a retrospective chart review conducted at the Level II Trauma Center. PATIENTS AND METHODS: Adult patients who sustained trauma and underwent chest and/or abdominal computed tomography (CT) scans and angiography were included in the study. Demographic and clinical information was extracted from patients who had CT scan and angiography. Extravasation of contrast material on CT scan and angiography was considered positive for ongoing internal bleeding. RESULTS: From January 2002 through July 2007, a total of 113 consecutive patients were included in this study. Forty-seven patients had evidence of ongoing internal bleeding (41.6%; 95% confidence interval: 32.4%, 51.2%). When comparing patients with and without ongoing bleeding, these two groups were similar in their gender, race, pulse, injury severity score and shock index. However, bleeding patients were typically older [mean (standard deviation): 44.5 (20.5) vs 37.3 (19.1) years; P = 0.051], had a lower initial SBP [116.2 (36.0) vs 130.0 (30.4) mmHg; P = 0.006] and had a higher Glasgow coma scale (GCS) [13.1 (4.0) vs 12.1 (4.4); P = 0.09]. From a multivariate logistic regression analysis, older age (P = 0.046) and lower SBP (P = 0.01) were significantly associated with bleeding, when controlled for gender, race and GCS. Among the 47 patients with ongoing bleeding, only seven patients (15%) had a SBP lower than 90 mmHg and 25 patients (53%) had a SBP higher than or equal to 120 mmHg. The spleen was the most frequently injured organ identified with active bleeding. CONCLUSIONS: Initial SBP cannot predict the ongoing internal bleeding. PMID- 21633567 TI - Retraction notice. PMID- 21633566 TI - Prediction of difficult intubations using conventional indicators: Does rapid sequence intubation ease difficult intubations? A prospective randomised study in a tertiary care teaching hospital. AB - BACKGROUND: Endotracheal intubations performed in the Emergency Department. AIMS: To assess whether conventional indicators of difficult airway can predict a difficult intubation in the Emergency Setting and to investigate the effect of rapid sequence intubation (RSI) on ease of intubation. SETTINGS AND DESIGN: A prospective randomized study was designed involving 60 patients requiring intubation, over a period of 4 months. MATERIALS AND METHODS: Demographic profile, details of methods used, airway assessment, ease of intubation, and Cormack and Lehane score were recorded. Airway assessment score and ease of intubation criteria were devised and assessed. STATISTICAL ANALYSIS: Descriptive statistical analysis was carried out. Chi-square/2 * 2, 2 * 3, 3 * 3, Fisher Exact test have been used to find the significance of study parameters on categorical scale between two or more groups. RESULTS: Patients with a Mallampatti score of three or four were found to have worse laryngoscopic views (Cormack-Lehane score, 3 or 4). Of all airway indicators assessed, an increased Mallampatti score was found to have significant correlation with increased difficulty in intubation. The use of RSI was associated with better laryngoscopic views, and easier intubations. CONCLUSIONS: An airway assessment using the Mallampatti score is invaluable as a tool to predict a difficult airway and should be performed routinely if possible. RSI aids intubation ease. If not otherwise contraindicated, it should be performed routinely for all intubations in the ED. PMID- 21633568 TI - Analysis of the pattern of maxillofacial fractures in north western of Iran: A retrospective study. AB - BACKGROUND: Maxillofacial fractures can lead to substantial long-term functional, esthetic and psychological complications. AIM: The aim of this study is to evaluate these injuries in a Turkish Iranian population. MATERIALS AND METHODS: A retrospective study of 170 patients with 210 maxillofacial fractures admitted to the emergency department of a central referral emergency hospital in the area over a 5 year period is presented. Patients' data included demographic information, etiology, site and associated injuries and complications. RESULTS: Road traffic accident was the commonest cause (40%) and the age group of 21-30 comprised the biggest group (30%). Mandibular fractures outnumbered midface fractures (150 vs. 60). Ramus (21.5%) and zygoma (26.5%) were the commonest fracture regions respectively in mandible and midface. Male: female ratio was 3.8:1 Almost half of patients (46%) had sustained associated injuries most of which was soft tissue laceration of the face (17.5%). 22 patient (13%) had associated complication and the hemorrhage was the commonest form of that (9%). CONCLUSION: It seems that road traffic accidents continue to be the leading cause of maxillofacial fractures and there is an urgent need to implement enhanced regulations and monitoring on motor vehicular traffic. PMID- 21633569 TI - Current practice and the role of the CT in the management of penetrating liver injuries at a Level I trauma center. AB - BACKGROUND: The liberal utilization of computed tomography (CT) has significantly contributed to overall improvements in trauma care. However, the role and the current practice of the CT examinations in the management of patients with penetrating liver injuries are scantily documented. AIMS: This study was aimed to assess the current practice and the role of the admission and follow-up CT in patients with penetrating liver injuries. SETTING AND DESIGN: This is a retrospective study at a Level I trauma center. Study period is from 01/2005 to 12/2007. METHODS: All patients with penetrating liver injuries were analyzed. RESULTS: Overall, 178 patients with penetrating liver injuries were assessed. A total of 123 (69.1%) patients underwent emergent laparotomy without preoperative CT due to signs of peritonitis (47.8%), hypotension (16.3%), or a GCS of <=8 (5.1%). In this group of patients, no nontherapeutic laparotomy occurred. The remaining 55 (30.9%) patients underwent CT scan evaluation on admission. Of these, 54.5% were selected for nonoperative management (NOM). Sensitivity and specificity of the admission CT to predict a positive laparotomy was 95.7% and 90.6%, respectively. Overall, 80.6% of isolated liver injuries were successfully managed nonoperatively. Thirty-three (18.5%) patients died within 72 h. In the remaining 145 patients, 33 liver-related complications occurred in 17.2% (25 of 145) of patients. CONCLUSION: Two-thirds of patients with penetrating liver injuries require emergent laparotomy, mainly due to associated injuries. The remaining one-third of patients, however, is amenable for an admission CT, which reliably predicts successful NOM. Moderate or severe injuries require follow-up CT because of the high incidence of asymptomatic liver-related complications. PMID- 21633570 TI - Determinants of mortality in trauma patients following massive blood transfusion. AB - AIM: This study was designed to find out the factors influencing mortality in trauma patients receiving massive blood transfusion (MBT). MATERIALS AND METHODS: Records of all patients admitted during December 2007 to November 2008 at a Level I Trauma Center emergency and who underwent massive transfusion (>=10 units of packed red cells in 24 h) were retrospectively analyzed. Death during the hospital stay was considered as the study outcome and various demographic, laboratory, and clinical parameters were included as its potential determinants. STATISTICAL ANALYSIS: Bivariate and multivariate logistic regression analyses were done to identify the risk factors associated with mortality. RESULTS: Of the 4054 transfused patients who were admitted to the trauma center during the study period, 71 (1.8%) patients underwent massive transfusion. Of this, there were 37 survivors and 34 nonsurvivors (48%). The median overall ISS was 27 (22-34). The patients who died had shorter mean length of hospital stay, shorter mean duration of intensive care unit (ICU) stay, and low admission Glasgow Coma Scale (GCS) compared to the survivors (P < 0.01). The mean prothrombin time (PT) and the mean activated partial thromboplastin time was significantly high (P < 0.01) among nonsurvivors. Total leukocyte count (TLC >= 10,000 cells/cubic mm), GCS <= 8, the presence of coagulopathy and major vascular surgery were the four independent determinants of mortality in multivariate logistic regression analysis. The FFP:PRBC (fresh frozen plasma:packed red cells) ratio and PC:PRBC (platelet concentrate:packed red cells) ratio calculated in our study was not statistically significant in correlation to the in hospital mortality. CONCLUSIONS: Overall mortality among the MBT patients was comparable with the studies in the literature. Mortality is not affected by the amount of packed red cells given in the first 12 h and the total number of packed red cells transfused. Prospective studies are required to further validate the determinants of mortality and establish guidelines for MBT. PMID- 21633571 TI - The impact of antiplatelet therapy on pelvic fracture outcomes. AB - INTRODUCTION: Despite increasing use of antiplatelet agents (APA), little is known regarding the effect of these agents on the orthopedic trauma patient. This study reviews clinical outcomes of patients with pelvic fractures (Pfx) who were using pre-injury APA. Specifically, we focused on the influence of APA on postinjury bleeding, transfusions, and outcomes after Pfx. METHODS: Patients with Pfx admitted during a 37-month period beginning January 2006 were divided into APA and non-APA groups. Pelvic injuries were graded using pelvic fracture severity score (PFSS)-a combination of Young-Burgess (pelvic ring), Letournel Judet (acetabular), and Denis (sacral fracture) classifications. Other clinical data included demographics, co-morbid conditions, medications, injury severity score (ISS), associated injuries, morbidity/mortality, hemoglobin trends, blood product use, imaging studies, procedures, and resource utilization. Multivariate analyses for predictors of early/late transfusions, pelvic surgery, and mortality were performed. RESULTS: A total of 109 patients >45 years with Pfx were identified, with 37 using preinjury APA (29 on aspirin [ASA], 8 on clopidogrel, 5 on high-dose/scheduled non-steroidal anti-inflammatory agents [NSAID], and 8 using >1 APAs). Patients in the APA groups were older than patients in the non APA group (70 vs. 63 years, P < 0.01). The two groups were similar in gender distribution, PFSS and ISS. Patients in the APA group had more comorbidities, lower hemoglobin levels at 24 h, and received more packed red blood cell (PRBC) transfusions during the first 24 h of hospitalization (all, P < 0.05). There were no differences in platelet or late (>24 h) PRBC transfusions, blood loss/transfusions during pelvic surgery, lengths of stay, post-ED/discharge disposition, or mortality. In multivariate analysis, predictors of early PRBC transfusion included higher ISS/PFSS, pre-injury ASA use, and lower admission hemoglobin (all, P < 0.03). Predictors of late PRBC transfusion included the number of complications, gender, PFSS, and any APA use (all, P < 0.05). Mortality was associated with pelvic hematoma/contrast extravasation on imaging, number of complications, and higher PFSS/ISS (all, P < 0.04). CONCLUSIONS: Results of this study support the contention that preinjury use of APA does not independently affect morbidity or mortality in trauma patients with Pfx. Despite no clinically significant difference in early postinjury blood loss, pre-injury use of APA was associated with increased likelihood of receiving PRBC transfusion within 24 h of admission. Furthermore, multivariate analyses demonstrated that among different APA, only preinjury ASA (vs. clopidogrel or NSAID) was associated with early PRBC transfusions. Late transfusion was associated with the use of any APA, complications, higher PFSS, and need for pelvic surgery. PMID- 21633572 TI - Management of ureteral calculi and medical expulsive therapy in emergency departments. AB - INTRODUCTION: Ureteral stones are a common problem in daily emergency department practice. Patients may be offered medical expulsive therapy (MET1) to facilitate stone expulsion and this should be offered as a treatment for patients with distal ureteral calculi, who are amenable to waiting management. Emergency department clinicians and family practitioners are often in the front line regarding the diagnosis and treatment of symptomatic nephrolithiasis and this commentary is dedicated to them because their decisions directly influence the outcome of the acute stone episode and appropriate referral patterns. MATERIALS AND METHODS: The aim of this systematic review and meta-analysis was to understand the role of MET in the treatment of obstructing ureteral calculi. A bibliographic search covering the period from January 1980 to March 2010 was conducted in PubMed, MEDLINE and EMBASE. The searches were restricted to publications in English. This analysis is based on the 21 studies that fulfilled the predefined inclusion criteria. RESULTS: A metaregression analysis of expulsion time showed a statistically significant advantage in the experimental group, in which the mean expulsion time was 6.2 days compared to 10.3 days in controls. The treatment effect on expulsion rate (P = 0.53) was partially lost as the size of the stones decreased because of the high spontaneous expulsion rate of small stones and the expulsion time was not influenced by pharmacological treatment (P = 0.76) if the stone size was smaller than 5 mm. Analysis of the tamsulosin database. A total of 1283 participants were included in the 17 studies. These studies showed that compared to standard therapy or placebo, tamsulosin had significant benefits, being associated with both a higher stone expulsion rate (P < 0.001) and reduction of the expulsion time (P = 0.02). Reductions in the need for analgesic therapy, hospitalization and surgery are also shown. Analysis of the nifedipine database. The number of participants in each trial ranged from 25 to 70. Compared to standard therapy, the use of nifedipine significantly improved the spontaneous stone expulsion rate (P < 0.001). The mean expulsion time was slightly, but not statistically significantly, different (P = 0.19) between the treatment and control groups. A possible benefit of nifedipine, in terms of significantly reducing the doses of analgesics required, was reported in three studies. There was no difference between the tamsulosin- and nifedipine-treated groups with regard to expulsion time (P = 0.17) or expulsion rate (P = 0.79). CONCLUSIONS: Despite all its advantages, MET is rarely used, representing a failure of the translation of medical science into practice. These data raise concerns not only about the quality of care of patients who could benefit from resolution of stones without anaesthetic and surgical risks but also with regard to potential cost savings. MET should be offered as a treatment for patients with distal ureteral calculi who are amenable to a waiting management. PMID- 21633573 TI - Treatment of major vein injury with the hemostatic fleece TachoSil by interposing a peritoneal patch to avoid vein thrombosis: A feasibility study in pigs. AB - BACKGROUND: Vein lacerations in awkward locations are difficult to repair and carry high mortality. The hemostatic fleece, TachoSil, is effective in preventing intraoperative bleeding in different settings, but has not been recommended for use in large vein injury. TachoSil with a peritoneal patch interposed to avoid vein thrombosis has been reported as a method to obtain hemostasis in vein laceration, but further studies of this method are needed. MATERIALS AND METHODS: A 1.5 * 1 cm defect was created in the vena cava in five pigs. A 26 * 32 mm peritoneal patch was applied on the coagulant side of a 48 * 48 mm TachoSil sheet, and used to cover the defect. Light compression with a wet sponge was applied for 3 min. No vascular suturing was performed. RESULTS: Successful hemostasis was obtained in four out of the five pigs although the minimum TachoSil gluing zone surrounding the peritoneal patch was only 0-2 mm. The fifth pig died of hemorrhage 30 min after surgery due to a 4-mm stretch with no TachoSil gluing zone outside the peritoneal patch. At six days postoperatively the peritoneal patch was well integrated into the vein wall. After 28 days, the peritoneal patch was almost indiscernible from surrounding vein endothelium. CONCLUSIONS: Vein wall defects can be repaired using TachoSil with a peritoneal patch interposed to prevent contact between the thrombogenic TachoSil sheet and the vein lumen. An adequate TachoSil gluing zone all around the patch is essential. PMID- 21633574 TI - Neuroprotective effect of wormwood against lead exposure. AB - BACKGROUND: Lead poisoning is a potential factor in brain damage, neurochemical dysfunction and severe behavioral problems. Considering this effect, our study was carried out to investigate the effects of wormwood to restore enzymes activities, lipid peroxidation and behavioral changes induced by lead. METHODS: Thirty Wistar rats were divided into five groups (n = 6 in each group): three groups exposed to 750 ppm of lead acetate in the drinking water for 11 weeks and two groups as control. Aqueous wormwood extract (200 mg/kg body weight) was administrated to intoxicated (Pb(-)+A.AB) and control groups (A.AB) for four supplemental weeks. Activities of acetylcholinesterase (AchE), monoamine oxidase (MAO) and thiobarbituric acid-reactive substances (TBARS) level were determined in the hypothalamus, hippocampus, cortex and striatum of male rats and the grooming and locomotors activity were defined in all groups. RESULTS: The intoxicated group (Pb) has a significantly increased TBARS value compared with the control in all regions (P < 0.05) and, after treatment with the wormwood extract, a significant reduction was noted. The enzyme activity decreased significantly (P < 0.05) in the Pb group compared with the control, essentially for the hippocampus (AchE: -57%, MAO: -41%) and the striatum (AchE: -43%, MAO: 51%). After wormwood extract administration, the AchE and MAO activity were significantly increased in all brain regions compared with the Pb group (P < 0.05). The behavioral test (locomotors and grooming test) indicates a significant hyperactivity in the Pb group compared with the control group. After treatment with wormwood extract, the Pb(-)+A.Ab indicates a lower activity compared with Pb. CONCLUSION: These data suggest that wormwood extract may play a very useful role in reduction of the neurotoxicological damage induced by lead. PMID- 21633575 TI - Pediatric cardiac emergencies: Children are not small adults. AB - Compared with adults, cardiac emergencies are infrequent in children and clinical presentation is often quite variable. In adults, cardiac emergencies are most commonly related to complications of coronary artery disease; however, in pediatric cases, the coronaries are only rarely the underlying problem. Pediatric cardiac emergencies comprise a range of pathology including but not limited to undiagnosed congenital heart disease in the infant; complications of palliated congenital heart disease in children; arrhythmias related to underlying cardiac pathology in the teenager and acquired heart disease. The emergency room physician and pediatric intensivist will usually be the first and second lines of care for pediatric cardiac emergencies and thus it is imperative that they have knowledge of the diverse presentations of cardiac disease in order to increase the likelihood of delivering early appropriate therapy and referral. The objective of this review is to outline cardiac emergencies in the pediatric population and contrast the presentation with adults. PMID- 21633576 TI - Heparin-induced thrombocytopenia. AB - In the last 7 decades heparin has remained the most commonly used anticoagulant. Its use is increasing, mainly due to the increase in the number of vascular interventions and aging population. The most feared complication of heparin use is heparin-induced thrombocytopenia (HIT). HIT is a clinicopathologic hypercoagulable, procoagulant prothrombotic condition in patients on heparin therapy, and decrease in platelet count by 50% or to less than 100,000, from 5 to 14 days of therapy. This prothrombotic hypercoagulable state in HIT patient is due to the combined effect of various factors, such as platelet activation, mainly the formation of PF4/heparin/IgG complex, stimulation of the intrinsic factor, and loss of anticoagulant effect of heparin. Diagnosis of HIT is done by clinical condition, heparin use, and timing of thrombocytopenia, and it is confirmed by either serotonin release assay or ELISA assay. Complications of HIT are venous/arterial thrombosis, skin gangrene, and acute platelet activation syndrome. Stopping heparin is the basic initial treatment, and Direct Thrombin Inhibitors (DTI) are medication of choice in these patients. A few routine but essential procedures performed by using heparin are hemodialysis, Percutaneous Coronary Intervention, and Cardiopulmonary Bypass; but it cannot be used if a patient develops HIT. HIT patients with unstable angina, thromboembolism, or indwelling devices, such as valve replacement or intraaortic balloon pump, will require alternative anticoagulation therapy. HIT can be prevented significantly by keeping heparin therapy shorter, avoiding bovine heparin, using low-molecular weight heparin, and stopping heparin use for flush and heparin lock. PMID- 21633577 TI - Transfusion protocol in trauma. AB - Blood and blood components are considered drugs because they are used in the treatment of diseases. As with any drug, adverse effects may occur, necessitating careful consideration of therapy. Like any other therapeutic decision, the need for transfusion should be considered on the basis of risks and benefits and alternative treatments available to avoid over- and under-transfusion. This review is focused on the blood transfusion protocol in trauma patients with hemorrhagic shock. Besides, issues related to emergency and massive transfusion have also been elaborated. We conducted a comprehensive MEDLINE search and reviewed the relevant literature, with particular reference to emergency medical care in trauma. PMID- 21633578 TI - A primer on burn resuscitation. AB - Since the early 1900s, the scope of burn resuscitation has evolved dramatically. Due to various advances in pre-hospital care and training, under-resuscitation of patients with severe burns is now relatively uncommon. Over-resuscitation, otherwise known as "fluid creep", has emerged as one of the most important problems during the initial phases of burn care over the past decade. To avoid the complications of over-resuscitation, careful hourly titration of fluid rates based on compilation of various clinical end points by a bedside provider is vital. The aim of this review is to provide a practical approach to the resuscitation of severely burned patients. PMID- 21633579 TI - Management of liver trauma in adults. AB - The liver is one of the most commonly injured organs in abdominal trauma. Recent advancements in imaging studies and enhanced critical care monitoring strategies have shifted the paradigm for the management of liver injuries. Nonoperative management of both low- and high-grade injuries can be successful in hemodynamically stable patients. Direct suture ligation of bleeding parenchymal vessels, total vascular isolation with repair of venous injuries, and the advent of damage control surgery have all improved outcomes in the hemodynamically unstable patient population. Anatomical resection of the liver and use of atriocaval shunt are rarely indicated. PMID- 21633580 TI - Dengue shock. AB - Shock syndrome is a dangerous complication of dengue infection and is associated with high mortality. Severe dengue occurs as a result of secondary infection with a different virus serotype. Increased vascular permeability, together with myocardial dysfunction and dehydration, contribute to the development of shock, with resultant multiorgan failure. The onset of shock in dengue can be dramatic, and its progression relentless. The pathogenesis of shock in dengue is complex. It is known that endothelial dysfunction induced by cytokines and chemical mediators occurs. Diagnosis is largely clinical and is supported by serology and identification of viral material in blood. No specific methods are available to predict outcome and progression. Careful fluid management and supportive therapy is the mainstay of management. Corticosteroids and intravenous immunoglobulins are of no proven benefit. No specific therapy has been shown to be effective in improving survival. PMID- 21633581 TI - Gastrothorax or tension pneumothorax: A diagnostic dilemma. AB - Gastrothorax, a rare complication following thoracoabdominal aortic aneurysm repair, is reported. The clinical features of a gastrothorax and tension pneumothorax are similar and thus, a gastrothorax can masquerade as a tension pneumothorax. The diagnosis is made by a high level of clinical suspicion, chest X-ray shows a distended stomach with air fluid levels and a computerised tomography is useful in assessing the diaphragm and establishing the positions of the various intra-abdominal organs. Also, the risk of an intercostal drainage tube placement and the role of nasogastric tube in avoiding the development of a tension gastrothorax is highlighted. PMID- 21633582 TI - Retroperitoneal vascular malformation mimicking incarcerated inguinal hernia. AB - A 30-year-old man presented to the Department of Surgery with a painful groin swelling on right side. Exploration revealed a reddish-blue hemangiomatous mass in the scrotum extending through inguinal canal into the retroperitoneum. On further dissection swelling was found to be originating from right external iliac vein. The swelling was excised after ligating all vascular connections. The histopathological examination of excised mass confirmed the diagnosis of venous variety of vascular malformation. This is the first reported case of vascular malformation arising from retroperitoneum and extending into inguinoscrotal region, presenting as incarcerated inguinal hernia. PMID- 21633583 TI - A case of organophosphate poisoning presenting with seizure and unavailable history of parenteral suicide attempt. AB - Organophosphate (OP) poisoning is common in India. Only few case reports of parenteral OP poisoning have been described. We report a case of self-injected methyl parathion poisoning, presenting after four days with seizure, altered sensorium, and respiratory distress which posed a diagnostic and therapeutic dilemma. Despite nonavailability of history of OP poisoning, he was treated based on suspicion and showed a good clinical response to treatment trial with atropine and pralidoxime, and had a successful recovery. Atypical presentations may be encountered following parenteral administration of OP poison, and even a slight suspicion of this warrants proper investigations and treatment for a favorable outcome. Persistently low plasma cholinesterase level is a useful marker for making the diagnosis. PMID- 21633584 TI - Adhesive bowel obstruction? Not always. AB - A 58-year-old man presented acutely with features of post-surgical adhesive small bowel obstruction. Following an unsuccessful trial of conservative management, computed tomography (CT) of the abdomen was performed. This revealed a mass in the ileocaecal region, for which he underwent a subsequent right hemicolectomy. Histology revealed diffuse B-cell Non-Hodgkin's lymphoma of the terminal ileum. Confounding obstructive lesion of the intestine in patients with a history of previous laparotomy is extremely uncommon. Early high resolution imaging may predict diagnosis and consolidate clinical management plans. PMID- 21633585 TI - Liver transplantation for hepatic trauma: Discussion about a case and its management. AB - Liver transplant for trauma is a rare condition with 19 cases described in the literature. We report the case of a 16-year-old patient who suffered a gradeV liver injury with a vena cava tear after a car crash. After a computerized tomography (CT) scan, the patient was directly sent to the operating room where the surgeon performed a right hepatectomy extended to segment IV with a venous repair under discontinued hilar clamping. On day five, the patient developed acute liver failure and was put on an emergency transplant waiting list. He had a successful liver transplant 2 days later. Fifteen months after his transplant, the patient is alive and asymptomatic. This case report focuses on the patient's initial management, the importance of damage control surgery and the circumstances which finally led to the transplant. PMID- 21633586 TI - Glycopyrrolate in toxic exposure to ammonia gas. AB - Ammonia (NH(3)) is a highly water-soluble, colorless, irritant gas with a unique pungent odor. Liquid ammonia stored under high pressure is still widely used for refrigeration in cold stores used for storing grains. Severe toxicity may occur following accidental exposure. We report an interesting case of accidental exposure to ammonia treated with glycopyrrolate along with other supportive measures. PMID- 21633587 TI - Traumatic anterior abdominal wall hernia: A report of three rare cases. AB - Traumatic abdominal wall hernia is a rare condition that can follow any blunt trauma. Associated intra-abdominal injuries are infrequent. In this study, we are reporting three cases, diagnosed as abdominal wall hernia associated with herniation of bowel loops due to blunt trauma. In one case, injury of the herniated bowel was seen. In western medical literature, only few cases have been reported especially with intra-abdominal injuries. PMID- 21633588 TI - Backstabbing. PMID- 21633589 TI - Self-inflicted intracranial self-injury. PMID- 21633590 TI - Injuries due to fall make summer time power outages a potential public health issue. PMID- 21633591 TI - An arrowhead found incidentally in the chest during workup for unrelated disease after 22-years of initial injury. PMID- 21633592 TI - Recent advances in celiac disease. AB - Celiac disease now affects about one person in a hundred in Europe and North America. In this review, we consider a number of important and exciting recent developments, such as clinical associations, HLA-DQ2 and HLA-DQ8 predispositions, the concept of potential celiac disease, the use of new imaging/endoscopy techniques, and the development of refractory disease. This review will be of use to all internists, pediatricians and gastroenterologists. PMID- 21633593 TI - Management of liver cirrhosis between primary care and specialists. AB - This article discusses a practical, evidence-based approach to the diagnosis and management of liver cirrhosis by focusing on etiology, severity, presence of complications, and potential home-managed treatments. Relevant literature from 1985 to 2010 (PubMed) was reviewed. The search criteria were peer-reviewed full papers published in English using the following MESH headings alone or in combination: "ascites", "liver fibrosis", "cirrhosis", "chronic hepatitis", "chronic liver disease", "decompensated cirrhosis", "hepatic encephalopathy", "hypertransaminasemia", "liver transplantation" and "portal hypertension". Forty nine papers were selected based on the highest quality of evidence for each section and type (original, randomized controlled trial, guideline, and review article), with respect to specialist setting (Gastroenterology, Hepatology, and Internal Medicine) and primary care. Liver cirrhosis from any cause represents an emerging health issue due to the increasing prevalence of the disease and its complications worldwide. Primary care physicians play a key role in early identification of risk factors, in the management of patients for improving quality and length of life, and for preventing complications. Specialists, by contrast, should guide specific treatments, especially in the case of complications and for selecting patient candidates for liver transplantation. An integrated approach between specialists and primary care physicians is essential for providing better outcomes and appropriate home care for patients with liver cirrhosis. PMID- 21633594 TI - Gastroenterology training in Latin America. AB - Latin America is characterized by ethnic, geographical, cultural, and economic diversity; therefore, training in gastroenterology in the region must be considered in this context. The continent's medical education is characterized by a lack of standards and the volume of research continues to be relatively small. There is a multiplicity of events in general gastroenterology and in sub disciplines, both at regional and local levels, which ensure that many colleagues have access to information. Medical education programs must be based on a clinical vision and be considered in close contact with the patients. The programs should be properly supervised, appropriately defined, and evaluated on a regular basis. The disparity between the patients' needs, the scarce resources available, and the pressures exerted by the health systems on doctors are frequent cited by those complaining of poor professionalism. Teaching development can play a critical role in ensuring the quality of teaching and learning in universities. Continuing professional development programs activities must be planned on the basis of the doctors' needs, with clearly defined objectives and using proper learning methodologies designed for adults. They must be evaluated and accredited by a competent body, so that they may become the basis of a professional regulatory system. The specialty has made progress in the last decades, offering doctors various possibilities for professional development. The world gastroenterology organization has contributed to the speciality through three distinctive, but closely inter-related, programs: Training Centers, Train the-Trainers, and Global Guidelines, in which Latin America is deeply involved. PMID- 21633596 TI - Double-balloon-enteroscopy-based endoscopic retrograde cholangiopancreatography in post-surgical patients. AB - AIM: To evaluate double balloon enteroscopy (DBE) in post-surgical patients to perform endoscopic retrograde cholangiopancreatography (ERCP) and interventions. METHODS: In 37 post-surgical patients, a stepwise approach was performed to reach normal papilla or enteral anastomoses of the biliary tract/pancreas. When conventional endoscopy failed, DBE-based ERCP was performed and standard parameters for DBE, ERCP and interventions were recorded. RESULTS: Push enteroscopy (overall, 16 procedures) reached enteral anastomoses only in six out of 37 post-surgical patients (16.2%). DBE achieved a high rate of luminal access to the biliary tract in 23 of the remaining 31 patients (74.1%) and to the pancreatic duct (three patients). Among all DBE-based ERCPs (86 procedures), 21/23 patients (91.3%) were successfully treated. Interventions included ostium incision or papillotomy in 6/23 (26%) and 7/23 patients (30.4%), respectively. Biliary endoprosthesis insertion and regular exchange was achieved in 17/23 (73.9%) and 7/23 patients (30.4%), respectively. Furthermore, bile duct stone extraction as well as ostium and papillary dilation were performed in 5/23 (21.7%) and 3/23 patients (13.0%), respectively. Complications during DBE-based procedures were bleeding (1.1%), perforation (2.3%) and pancreatitis (2.3%), and minor complications occurred in up to 19.1%. CONCLUSION: The appropriate use of DBE yields a high rate of luminal access to papilla or enteral anastomoses in more than two-thirds of post-surgical patients, allowing important successful endoscopic therapeutic interventions. PMID- 21633595 TI - Silybin and the liver: from basic research to clinical practice. AB - Herbal products are increasingly used, mainly in chronic liver disease. Extracts of milk thistle, Silymarin and silybin, are the most prescribed natural compounds, with different indications, but with no definitive results in terms of clinical efficacy. This review analyzes the available studies on the effects of the purified product silybin, both as a free and a conjugated molecule, on liver cells or on experimentally induced liver damage, and in patients with liver disease. We searched PUBMED for articles pertaining to the in vitro and in vivo effects of silybin, its antifibrotic, anti-inflammatory, and antioxidant properties, as well as its metabolic effects, combined with the authors' own knowledge of the literature. Results indicate that the bioavailability of silybin phytosome is higher than that of silymarin and is less influenced by liver damage; silybin does not show significant interactions with other drugs and at doses < 10 g/d has no significant side effects. Experimental studies have clearly demonstrated the antifibrotic, antioxidant and metabolic effects of silybin; previous human studies were insufficient for confirming the clinical efficacy in chronic liver disease, while ongoing clinical trials are promising. On the basis of literature data, silybin seems a promising drug for chronic liver disease. PMID- 21633597 TI - (-)-Epigallocatechin-3-gallate inhibits VEGF expression induced by IL-6 via Stat3 in gastric cancer. AB - AIM: To demonstrate that (-)-Epigallocatechin-3-gallate (EGCG) inhibits vascular endothelial growth factor (VEGF) expression and angiogenesis induced by interleukin-6 (IL-6) via suppressing signal transducer and activator of transcription 3 (Stat3) activity in gastric cancer. METHODS: Human gastric cancer (AGS) cells were treated with IL-6 (50 ng/mL) and EGCG at different concentrations. VEGF, total Stat3 and activated Stat3 protein levels in the cell lyses were examined by Western blotting, VEGF protein level in the conditioned medium was measured by enzyme-linked immunosorbent assay, and the level of VEGF mRNA was evaluated by reverse transcription polymerase chain reaction (RT-PCR). Stat3 nuclear translocation was determined by Western blotting with nuclear extract, and Stat3-DNA binding activity was examined with Chromatin immunoprecipitation (ChIP) assay. IL-6 induced endothelial cell proliferation was measured with 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazoliumbromide assay, in vitro angiogenesis was determined with endothelial cell tube formation assay in Matrigel, and IL-6-induced angiogenesis in vitro was measured with Matrigel plug assay. RESULTS: There was a basal expression and secretion of VEGF in AGS cells. After stimulation with IL-6, VEGF expression was apparently up regulated and a 2.4-fold increase was observed. VEGF secretion in the conditioned medium was also increased by 2.8 folds. When treated with EGCG, VEGF expression and secretion were dose-dependently decreased. IL-6 also increased VEGF mRNA expression by 3.1 folds. EGCG treatment suppressed VEGF mRNA expression in a dose dependent manner. EGCG dose-dependently inhibited Stat3 activation induced by IL 6, but did not change the total Stat3 expression. When treated with EGCG or AG490, VEGF expressions were reduced to the level or an even lower level in the tumor cells not stimulated with IL-6. However, PD98059 and LY294002 did not change VEGF expression induced by IL-6. EGCG inhibited Stat3 nucleus translocation, and Stat3-DNA binding activity was also markedly decreased by EGCG. Furthermore, EGCG inhibited IL-6 induced vascular endothelial cell proliferation and tube formation in vitro and angiogenesis in vitro. CONCLUSION: EGCG inhibits IL-6-induced VEGF expression and angiogenesis via suppressing Stat3 activity in gastric cancer, which has provided a novel mechanistic insight into the anti-angiogenic activity of EGCG. PMID- 21633598 TI - LBP and CD14 polymorphisms correlate with increased colorectal carcinoma risk in Han Chinese. AB - AIM: To explore the associations of polymorphisms of lipopolysaccharide binding protein (LBP), cluster of differentiation 14 (CD14), toll-like receptor 4 (TLR 4), interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha) with the colorectal carcinoma (CRC) risk in Han Chinese. METHODS: Polymorphisms of LBP (rs1739654, rs2232596, rs2232618), CD14 (rs77083413, rs4914), TLR-4 (rs5030719), IL-6 (rs13306435) and TNF-alpha (rs35131721) were genotyped in 479 cases of sporadic colorectal carcinoma and 486 healthy controls of Han Chinese in a case control study. Single-nucleotide polymorphisms (SNPs) between cases and controls were analyzed by unconditional logistic regression. RESULTS: GA and GG genotypes of LBP rs2232596 were associated with a significantly increased risk of CRC [odds ratio (OR) = 1.51, 95% confidence interval (CI) 1.15-1.99, P = 0.003; OR = 2.49, 95% CI 1.16-5.38, P = 0.016, respectively]. A similar association was also observed for the CG genotype of CD14 rs4914 (OR= 1.69, 95% CI 1.20-2.36, P = 0.002). In addition, a combination of polymorphisms in LBP rs2232596 and CD14 rs4914 led to a 3.4-fold increased risk of CRC (OR = 3.44, 95% CI 1.94-6.10, P = 0.000). CONCLUSION: This study highlights the LBP rs2232596 and CD14 rs4914 polymorphisms as biomarkers for elevated CRC susceptibility in the Chinese Han population. PMID- 21633599 TI - Utility of pancreatography for diagnosing autoimmune pancreatitis. AB - AIM: To identify pancreatographic findings that facilitate differentiating between autoimmune pancreatitis (AIP) and pancreatic cancer (PC) on endoscopic retrograde cholangiopancreatography (ERCP) and magnetic resonance cholangiopancreatography (MRCP). METHODS: ERCP findings of 48 AIP and 143 PC patients were compared. Diagnostic accuracies for AIP by ERCP and MRCP were compared in 30 AIP patients. RESULTS: The following ERCP findings suggested a diagnosis of AIP rather than PC. Obstruction of the main pancreatic duct (MPD) was more frequently detected in PC (P < 0.001). Skipped MPD lesions were detected only in AIP (P < 0.001). Side branch derivation from the narrowed MPD was more frequent in AIP (P < 0.001). The narrowed MPD was longer in AIP (P < 0.001), and a narrowed MPD longer than 3 cm was more frequent in AIP (P < 0.001). Maximal diameter of the upstream MPD was smaller in AIP (P < 0.001), and upstream dilatation of the MPD less than 5 mm was more frequent in AIP (P < 0.001). Stenosis of the lower bile duct was smooth in 87% of AIP and irregular in 65% of PC patients (P < 0.001). Stenosis of the intrahepatic or hilar bile duct was detected only in AIP (P = 0.001). On MRCP, diffuse narrowing of the MPD on ERCP was shown as a skipped non-visualized lesion in 50% and faint visualization in 19%, but segmental narrowing of the MPD was visualized faintly in only 14%. CONCLUSION: Several ERCP findings are useful for differentiating AIP from PC. Although MRCP cannot replace ERCP for the diagnostic evaluation of AIP, some MRCP findings support the diagnosis of AIP. PMID- 21633600 TI - Prospective randomized controlled trial investigating the type of sutures used during hepatectomy. AB - AIM: To determine whether absorbable sutures or non-absorbable sutures are better in preventing surgical site infection (SSI), in this paper we discuss the results of a randomized clinical trial which examined the type of sutures used during hepatectomy. METHODS: All hepatic resections performed from January 2007 to November 2008 at the Department of Surgery at Iizuka Hospital in Japan were included in this study. There were 125 patients randomly assigned to an absorbable sutures (Vicryl) group or non-absorbable sutures (Silk) group. RESULTS: SSI was observed in 13.6% (17/125) patients participating in this study, 11.3% in the Vicryl group and 15.8% in the Silk group. Incisional SSI including superficial and deep SSI, was observed in 8% of the Vicryl group and 9.5% of the Silk group. Organ/space SSI was observed in 3.2% of the Vicryl group and 6.0% of the Silk group. There were no significant differences, but among the patients with SSI, the period for recovery was significantly shorter for the Vicryl group compared to the Silk group. CONCLUSION: The incidence of SSI in patients receiving absorbable sutures and silk sutures is not significantly different in this randomized controlled study; however, the period for recovery in patients with SSI was significantly shorter for absorbable sutures. PMID- 21633601 TI - XPD Lys751Gln polymorphism and esophageal cancer risk: a meta-analysis involving 2288 cases and 4096 controls. AB - AIM: To evaluate the association between xeroderma pigmentosum group D (XPD), genetic polymorphism Lys751Gln and esophageal cancer risk. METHODS: We searched PubMed up to September 1, 2010 to identify eligible studies. A total of 10 case control studies including 2288 cases and 4096 controls were included in the meta analysis. Statistical analysis was performed with Review Manage version 4.2. Odds ratios (ORs) with 95% confidence intervals (CIs) were used to assess the strength of the association. RESULTS: The results suggested that there is no significant association between XPD Lys751Gln polymorphism and esophageal cancer susceptibility in the overall population. However, in subgroup analysis by histology type, a significant association was found between XPD Lys751Gln polymorphism and esophageal adenocarcinoma (for CC vs AA: OR = 1.25, 95% CI = 1.01-1.55, P = 0.05 for heterogeneity). CONCLUSION: Our meta-analysis suggested that XPD Lys751Gln polymorphism may be associated with increased risk of esophageal adenocarcinoma. PMID- 21633602 TI - Lapatinib-induced hepatitis: a case report. AB - Lapatinib is an inhibitor of the tyrosine kinases of human epidermal growth factor receptor type 2 (HER2) and epidermal growth factor receptor type 1, with clinical activity in HER2-positive metastatic breast cancer. We present here a 60 year-old patient with metastatic breast cancer who presented with jaundice and increased serum aminotransferase levels and who had been treated with lapatinib for the previous 14 days. Laboratory tests excluded other causes of acute liver injury. Liver biopsy revealed lesions compatible with drug-induced hepatotoxicity. Bilirubin and liver enzymes returned to normal within three months of lapatinib discontinuation. Lapatinib should be included among the causes of drug-induced hepatitis. PMID- 21633603 TI - Transarterial injection of H101 in combination with chemoembolization overcomes recurrent hepatocellular carcinoma. AB - Transcatheter arterial chemoembolization (TACE) has become the standard treatment modality for unresectable hepatocellular carcinoma (HCC). Nonetheless, the clinical outcomes in patients with unresectable HCC are often unsatisfactory, especially in those with recurrent HCC. H101, an E1B gene deleted adenovirus, is known to have a significant antitumor activity. In addition, local injection of H101 can enhance the effect of antitumor therapies (chemotherapy and radiotherapy). Transarterial H101 gene injection in combination with TACE may help to control refractory and recurrent HCC. In this study, we report a 55-year old patient with recurrent HCC which was treated with transarterial injection of H101 in combination with TACE, leading to a good clinical prognosis of the patient. PMID- 21633604 TI - Prophylactic antibiotics for variceal hemorrhage: clostridium difficile infection still can be a risk. AB - Bron et al presented a retrospective study regarding the prophylactic use of antibiotics for variceal hemorrhage. Antibiotics appeared to improve the survival rate of patients without increasing clostridium difficile infection (CDI). We argue against the conclusion of the authors and consider that this result may be simply due to concurrent use of metronidazole, a therapeutic agent against CDI. PMID- 21633605 TI - Welcome to the readers. PMID- 21633606 TI - Management of provoked seizure. AB - A provoked seizure may be due to structural damage (resulting from traumatic brain injury, brain tumor, stroke, tuberculosis, or neurocysticercosis) or due to metabolic abnormalities (such as alcohol withdrawal and renal or hepatic failure). This article is a part of the Guidelines for Epilepsy in India. This article reviews the problem of provoked seizure and its management and also provides recommendations based on currently available information. Seizure provoked by metabolic disturbances requires correction of the triggering factors. Benzodiazepines are recommended for treatment of seizure due to alcohol withdrawal; gabapentin for seizure seen in porphyria; and antiepileptic drugs (AED), that are not inducer of hepatic enzymes, in the seizures seen in hepatic dysfunction. In severe traumatic brain injury, with or without seizure, phenytoin (PHT) may be given for 7 days. In ischemic or hemorrhagic stroke one may individualize the AED therapy. In cerebral venous sinus thrombosis (CVST), AED may be prescribed if there is seizure or computed tomographic (CT) abnormalities or focal weakness; the treatment, in these cases, has to be continued for 1 year. Prophylactic AED is not recommended in cases of brain tumor and neurosurgical procedures and if patient is on an AED it can be stopped after 1 week. PMID- 21633607 TI - Treating children with attention-deficit/hyperactivity disorder and comorbid epilepsy. AB - Attention-deficit/hyperactivity disorder (ADHD) is one of common neurodevelopmental disorder often comorbid with epilepsy. There are no existing guidelines on how to manage these two conditions when they are comorbid. To identify relationship between epilepsy and ADHD and to know role of antiepileptics and safety of stimulant like methylphenidate in such conditions from existing literature, we searched articles published in clinical journals available online between 1990-2010, with these key words in medline:children, epilepsy, seizure, comorbid, ADHD, treatment. Relevant abstracts were further selected for their focus on current topic. Cross references were extracted. Finally relevant articles that included original research articles, reviews and abstracts of non-english literature were used. Children with epilepsy may manifest with symptoms of ADHD. Children with ADHD may develop epilepsy. Some antiepileptics like phenobarbitone, gabapentin, topiramate may not be helpful in controlling behavioral symptoms of ADHD. Stimulants are the main stay of pharmacotherapy for ADHD but there is risk of decreasing seizure threshold in children with comorbid epilepsy especially when their epilepsy is not well controlled. Existing evidence is not in favor of screening children with ADHD for EEG abnormality before starting stimulant therapy. PMID- 21633608 TI - Meniere's disease: Still a mystery disease with difficult differential diagnosis. AB - One hundred and forty-six years after its first description, the differential diagnosis of Meniere's disease remains very challenging. The aim of the present study is to review the current knowledge on the advantages and disadvantages of the new diagnostic methods for Meniere's disease. The importance of accurate diagnosis for primary healthcare systems is also discussed. An extensive search of the literature was performed in Medline and other available database sources. Information from electronic links and related books were also included. Controlled clinical studies, prospective cohort studies, retrospective cohort studies, cross-sectional studies, case reports, written guidelines, systematic reviews, and books were selected. The typical clinical triad of symptoms from the vestibular and cochlear systems (recurrent vertigo, fluctuating sensorineural hearing loss and tinnitus) is usually the key for clinical diagnosis. Glycerol dehydration test and electrocochleography are the main diagnostic tests in current practice, while vestibular evoked myogenic potentials may be used in disease staging. Imagine techniques are not specific enough to set alone the diagnosis of Meniere's disease, although they may be necessary to exclude other pathologies. Recently developed 3D MRI protocols can delineate the perilymphatic/endolymphatic spaces of the inner ear and aid diagnosis. Meniere's disease is a continuous problem for the patients and affects their quality of life. Taking into account the frequent nature of the disease in certain countries, efforts for reliable diagnosis, prompt referral, and successful management are undoubtedly cost-effective for healthcare systems. PMID- 21633609 TI - Long-term visual outcome in idiopathic intracranial hypertension. AB - OBJECTIVE: To characterize the course, outcome, and risk of relapse or late worsening in a clearly defined cohort of idiopathic intracranial hypertension (IIH) over a long period of follow-up. MATERIALS AND METHODS: Retrospective chart review of patients with definite IIH was evaluated at the Sree Chitra Tirunal Institute for Medical Sciences and Technology between 1998 and 2006. Patients' demographic data, clinical, neuro-ophthalmic examinations, and treatment details were abstracted. Patients were further categorized into three groups based on whether they improved, worsened, or relapsed on follow-up. Final visual outcome of each patient was defined according to grading of the worse eye at the last visit. Statistical analysis included t test to compare group means and chi-square test to compare proportions. RESULTS: Of the 43 women included, visual impairment was observed in 80 eyes (93%) at presentation and it was moderate to severe in 14%. The mean CSF opening pressure at presentation did not differ significantly in those with visual impairment compared to those with normal vision. Those having early severe visual impairment had significantly higher (P = 0.015) likelihood of severe visual impairment on last follow-up. Of the total, 34 patients (79%) improved, 4 (9.3%) relapsed on follow-up after period of stability, and 5 (11.6%) worsened over 56 months follow-up (range, 26-132 months). The groups were comparable, except those who improved were younger (P<0.05). At last examination, 9% had significant vision loss. CONCLUSION: IIH patients can have delayed worsening or relapses and about tenth of patients can have permanent visual loss early or late in the course of the disease. All patients with IIH need to be kept under long-term follow-up, with regular monitoring of visual functions. PMID- 21633610 TI - Intra-rater reliability of the modified Tardieu scale to quantify spasticity in elbow flexors and ankle plantar flexors in adult stroke subjects. AB - OBJECTIVES: The purpose of this study was to investigate Iintra-rater reliability of the Modified Tardieu Scale (MTS) in elbow flexors and ankle plantar flexors in adult subjects with stroke. MATERIALS AND METHODS: A total of 91 subjects with stroke participated in this test-retest study. Intra-rater reliability of the MTS was investigated by a qualified and trained physiotherapist for elbow flexors and ankle plantar flexors in two sessions. A rater was one who performed the procedure and an observer only records the angles so that the rater was blinded to findings. Outcome measures in this study were measurable components of MTS, which are angle of muscle reaction (R1), passive range of motion (R2), dynamic component (R2-R1), and quality of muscle reaction (grade 0 - 4) termed as MTS score. RESULTS: Intra-rater reliability of MTS was very good for R1, R2, R2-R1, and MTS score (ICC > 0.85, P<0.0001) across two sessions in elbow flexors and ankle plantar flexors. CONCLUSION: MTS is a reliable clinical tool for measurement of spasticity in the elbow flexors and ankle plantar flexors in adult subjects with stroke. PMID- 21633611 TI - Awareness of neurocysticercosis: A study from northwest India. AB - BACKGROUND: Neurocysticercosis (NCC) is a common cause of epilepsy in developing countries. In order to plan and implement prevention programs, it is essential to study the awareness of NCC. OBJECTIVE: To study the awareness of NCC among patients with NCC and compare with age- and gender-matched controls without NCC. SETTING AND DESIGN: Hospital based case-control study. MATERIALS AND METHODS: Two hundred and fourteen subjects were studied (109 NCC patients, and 105 age- and gender-matched controls without NCC). The participants were selected from neurology and medical wards of a tertiary referral hospital in northwest India. They were interviewed by trained medical interns using a questionnaire. RESULTS: 64.2% of the NCC patients and 19% of control group had heard about NCC (P < 0.001). Knowledge regarding organ affected by NCC in the NCC group was 61.4% and in the control group was 80% (P = 0.09). Only 12.9% of the NCC group and none in the control group identified tape worm as a causative agent for NCC (P = 0.092). Negative effects of NCC on marriage and social life were more often cited by the NCC group but in the control group it was towards education (P = 0.004). CONCLUSIONS: The awareness of NCC was poor in both the groups. Educational programs are needed to improve the awareness about NCC among the patients and the public. PMID- 21633612 TI - Sleep in Wilson's disease: Questionnaire based study. AB - OBJECTIVE: We proposed to detect sleep abnormalities in Wilson's disease, (WD) using sleep questionnaires. MATERIALS AND METHODS: Twenty-five patients (M:F = 18:7; age: 24.4 +/- 9.2 years) with WD and 24 controls (all males; age: 33.1 +/- 9.7 years) were recruited. They underwent phenotypic/magnetic resonance imaging (MRI) evaluation followed by administration of Pittsburg Sleep Quality Index (PSQI) and Epworth Sleepiness Scale (ESS) questionnaires. RESULTS: The mean age at presentation and diagnosis was 24.4 +/- 9.2 and 17.6 +/- 7.5 years, respectively. The duration of illness at diagnosis was 14 +/- 21.9 months. On PSQI, 15 patients with WD had abnormal PSQI scores of >5 as compared to 6 patients among the controls. The mean PSQI score was significantly more (P = 0.03) in patients compared to the controls. The PSQI worst scores were noted only in WD. Evaluation with ESS showed that three patients with WD scored >10/24, while two among the controls qualified for excessive daytime sleepiness. Overall, assessment by sleep questionnaires detected abnormality in 16 patients with WD as compared to 8 controls (P = 0.004). Subgroup analysis revealed that patients whose duration of illness was >8 years and who were on decoppering treatment had significantly lesser excessive daytime somnolence. CONCLUSIONS: Sleep disturbances were observed more often in WD than in controls. Better designed studies will provide a better understanding. PMID- 21633613 TI - Multidetector computed tomography angiography: Application in vertebral artery dissection. AB - BACKGROUND AND PURPOSE: Multidetector computed tomography angiography (MDCTA) is a minimally invasive radiological technique providing high-resolution images of the arterial wall and angiographic images of the lumen. We studied the radiological features of vertebral artery dissection (VAD) in a consecutive series of patients investigated for acute stroke and subarachnoid hemorrhage (SAH) in order to confirm and define the diagnostic features of VAD on MDCTA. PATIENTS AND METHODS: Review of patients identified prospectively over a 4-year period with VAD assessed by MDCTA was conducted. Radiological features of VAD on MDCTA were reanalyzed utilising previously reported criteria for VAD. RESULTS: Thirty-five patients (25 males, mean age 49.6 years) with a total of 45 dissected vertebral arteries were reviewed. MDCTA features of VAD included increased wall thickness in 44/45 (97.7%) arteries and increased total vessel diameter in 42/45 arteries (93.3%). All dissected arteries had either lumen stenosis (21/45) or associated segmental occlusion (24/45). An intimal flap was detected in 6/45 (13.3 %) vessels. Twenty-five patients had follow-up imaging, 14/32 vessels returned to normal, 4 showed improvement in stenosis but did not return to normal and 14 demonstrated no change. The majority of non-occluded vessels became normal or displayed improved patency. Only 4/17 occluded arteries demonstrated re establishment of flow. No adverse effects were recorded. CONCLUSIONS: MDCTA is a safe and reliable technique for the diagnosis of VAD. Increased wall thickness (97.7%) and increased vessel wall diameter (93.3%) were the most frequently observed features. PMID- 21633614 TI - Transition from curative to palliative care in cancer. PMID- 21633615 TI - Evaluation of Knowledge Among Interns in a Medical College Regarding Palliative Care in People Living with HIV/AIDS and the Impact of a Structured Intervention. AB - BACKGROUND: The evolving nature of palliative care and its renewed role in people living with HIV/AIDS (PLWHA) in the post-HAART (highly active anti-retroviral therapy) era warrants an evaluation of the present curriculum in medical under graduates. OBJECTIVES: The objectives are(1) to measure the existing knowledge regarding palliative care and its application to PLWHA among medical interns and (2) to measure the impact of a structured intervention on knowledge dimensions. DESIGN AND SETTING: Interventional repeated measures study. MATERIALS AND METHODS: A convenience sample of 106 interns in the medical college completed a pre-test assessment and a post-test assessment following a structured intervention for evaluation and comparison of knowledge over three dimensions which were (1) knowledge of palliative care and its application in PLWHA, (2) medical symptoms in PLWHA requiring palliative care and (3) psychosocial needs in PLWHA requiring palliative care. RESULTS: The mean scores on knowledge showed a consistent increase after the structured intervention and Student's t-test was significant across three dimensions of knowledge of palliative care and its application (t=9.12, P value <0.001), medical symptoms in PLWHA requiring palliative care (t=12.72, P value <0.001) and psychosocial needs in PLWHA (t=11.14, P value <0.001). CONCLUSION: In spite of the unique challenges presented by the varying course of illness in PLWHA and the variety of needs on the medical, psychosocial and family dimensions, a structured approach and an integrated course curriculum involving principles of both primary and palliative care principles will improve the efficiency of the undergraduate medical education program and enable delivery of effective palliative care interventions and improve quality of life in PLWHA. PMID- 21633616 TI - Evaluating the role of support group structures as vehicles of palliative care: giving productivity in the kanye care program in botswana. AB - AIMS: The article aims to explore, evaluate, and discuss the role associated with the presence or absence of caregivers' support groups in the Kanye care program in Botswana. OBJECTIVE: The objective of the study was to solicit the views, attitudes, and thinking of palliative caregivers on the role, and implication of the presence and absence of support groups as vehicles influencing care-giving productivity. MATERIALS AND METHODS: The study was explorative and descriptive in nature and used a qualitative design, involving 82 primary caregivers in focus group discussions and five community home-based care nurses in one- to-one interviews. RESULTS: THE SUPPORT GROUPS WERE FOUND TO PLAY THE FOLLOWING PIVOTAL ROLES: (1) facilitate forums of strengthening caregivers; (2) facilitate sharing and exchange of information among caregivers; (3) facilitate counselling, debriefings, and education among caregivers; (4) foster the spirit and feelings of togetherness among caregivers; and (5) act as advocacy fronts and an opportunity of pooling and mobilizing the resources. CONCLUSION: The article recommends to the Government and the nongovernmental organizations to assist all care programs to institute and facilitate the formation of caregivers' support groups. PMID- 21633617 TI - Awareness of palliative care among diploma nursing students. AB - BACKGROUND: The goal of palliative care is not to cure, but to provide comfort and maintain the highest possible quality of life for as long as life remains. The knowledge of nurses influences the quality of care provided to these patients. The present study aimed at identifying the level of knowledge and attitude of nursing students who are the future caretakers of patients, which helps to make recommendations in incorporating palliative care concepts in the nursing curriculum. OBJECTIVES: (1) To assess the level of knowledge of nursing students on palliative care; (2) To identify the attitude of nursing students towards palliative care; (3) To find the correlation between the knowledge and attitude of nursing students; (4) To find the association between nursing students' knowledge, attitude and selected demographic variables. MATERIALS AND METHODS: A correlative survey was carried out among 83 third-year Diploma Nursing students by using cluster sampling method from selected nursing schools of Udupi district. RESULTS: The data analyzed showed that the majority (51%) of them was in the age group of 21years and 92% of them were females. Only 43.4% of them were aware of the term palliative care and it was during their training period. The data showed that 79.5% of students had poor knowledge (6.4+/- 1.64) on palliative care and 92.8% of them had favorable attitude (56.7+/- 8.5) towards palliative care. The chi-square showed a significant association between knowledge and age (chi(2)=18.52,P<0.01) of the nursing students. CONCLUSION: Palliative care aspects should be incorporated in the diploma nursing curriculum. PMID- 21633618 TI - Impact of pain and palliative care services on patients. AB - BACKGROUND: Palliative care has become an emerging need of the day as the existing health-care facilities play only a limited role in the care of the chronically ill in the society. Patients with terminal illness in most cases spend their lives in the community among their family and neighbors, so there is the need for a multi disciplinary team for their constant care. Volunteers are primary care givers who originate normally from the same locality with local knowledge and good public contact through which they can make significant contributions in a team work by bridging the gap between the patient community and outside world. AIM: The present study has been undertaken to analyze the impact of palliative care services on patients by considering 51 variables. MATERIALS AND METHODS: The respondents of the study include 50 pain and palliative care patients selected at random from 15 palliative care units functioning in Ernakulam district. The analysis was made by using statistical techniques viz. weighted average method, Chi-square test, Friedman repeated measures analysis of variance on ranks and percentages. RESULTS: The study revealed that the major benefit of palliative care to the patients is the reduction of pain to a considerable extent, which was unbearable for them earlier. Second, the hope of patients could be maintained or strengthened through palliative care treatment. CONCLUSION: It is understood that the services of the doctors and nurses are to be improved further by making available their services to all the palliative care patients in a uniform manner. PMID- 21633620 TI - Clinical audit on documentation of anticipatory "not for resuscitation" orders in a tertiary Australian teaching hospital. AB - AIM: The purpose of this clinical audit was to determine how accurately documentation of anticipatory Not for Resuscitation (NFR) orders takes place in a major metropolitan teaching hospital of Australia. MATERIALS AND METHODS: Retrospective hospital-based study. Independent case reviewers using a questionnaire designed to study NFR documentation reviewed documentation of NFR in 88 case records. RESULTS: Prognosis was documented in only 40% of cases and palliative care was offered to two-third of patients with documented NFR. There was no documentation of the cardiopulmonary resuscitation (CPR) process or outcomes of CPR in most of the cases. Only in less than 50% of cases studied there was documented evidence to suggest that the reason for NFR documentation was consistent with patient's choices. CONCLUSION: Good discussion, unambiguous documentation and clinical supervision of NFR order ensure dignified and quality care to the dying. PMID- 21633619 TI - The Attitudes of Indian Palliative-care Nurses and Physicians to Pain Control and Palliative Sedation. AB - AIM: We wanted to assess Indian palliative-care nurses and physicians' attitudes toward pain control and palliative sedation. MATERIALS AND METHODS: From May to September 2008, we interviewed 14 physicians and 13 nurses working in different palliative-care programs in New Delhi, using a semi-structured questionnaire, and following grounded-theory methodology (Glaser and Strauss). RESULTS: The interviewees did not consider administration of painkillers in large doses an ethical problem, provided the pain killers are properly titrated. Mild palliative sedation was considered acceptable. The interviewees disagreed whether palliative sedation can also be deep and continuous. Arguments mentioned against deep continuous palliative sedation were the conviction that it may cause unacceptable side effects, and impedes basic daily activities and social contacts. A few interviewees said that palliative sedation may hasten death. CONCLUSION: Due to fears and doubts regarding deep continuous palliative sedation, it may sometimes be too easily discarded as a treatment option for refractory symptoms. PMID- 21633621 TI - Effects of Palliative Care Training Program on Knowledge, Attitudes, Beliefs and Experiences Among Student Physiotherapists: A Preliminary Quasi-experimental Study. AB - BACKGROUND: Physiotherapists play an inherent role in the multidisciplinary palliative care team. Existing knowledge, attitudes, beliefs and experiences influence their team participation in palliative care. AIMS: The objective of this study was to assess the changes in knowledge, attitudes, beliefs and experiences among student physiotherapists who attended a palliative care training program. SETTINGS AND DESIGN: Preliminary quasi-experimental study design, conducted at an academic institution. MATERIALS AND METHODS: Fifty-two student physiotherapists of either gender (12 male, 40 female) of age (20.51+/ 1.78 years) who attended a palliative care training program which comprised lectures and case examples of six-hours duration participated in this study. The study was performed after getting institutional approval and obtaining participants' written informed consent. The lecture content comprised WHO definition of palliative care, spiritual aspects of life, death and healing, principles, levels and models of palliative care, and role of physiotherapists in a palliative care team. The physical therapy in palliative care-knowledge, attitudes, beliefs and experiences scale (PTiPC-KABE Scale)- modified from palliative care attitudes scale were used for assessing the participants before and after the program. STATISTICAL ANALYSIS: Paired t-test and Wilcoxon signed rank test at 95% confidence interval using SPSS 11.5 for Windows. RESULTS: Statistically significant differences (P<0.05) were noted for all four subscales- knowledge (7.84+/-4.61 points), attitudes (9.46+/-8.06 points), beliefs (4.88+/ 3.29 points) and experiences (15.8+/-11.28 points) out of a total score of 104 points. CONCLUSIONS: The focus-group training program produced a significant positive change about palliative care in knowledge, attitudes, beliefs and experiences among student physiotherapists. PMID- 21633622 TI - Anticoagulant and Anti-thrombotic Treatments in the Management of Hematological Malignancies in a Home Care Program. AB - AIM: Anticoagulants (AC) and anti-platelet (AP) agents are widely administered to patients with hematological malignancies (HM). However, HM patients may be at high risk of bleeding and hemorrhagic complications, because of different form of coagulopathies and several degrees of thrombocytopenia. MATERIALS AND METHODS: A prospective evaluation of the use of anticoagulant and anti-thrombotic agents as well as of bleeding and thrombotic complications in a consecutive cohort of patients, which were followed during the first semester of 2010 by our home care service, was performed. In this regard, three pharmacological class of agents, such as oral anticoagulants (warfarin and acenocumarine), low molecular weight heparin (LMWH) and anti-platelet (AP) drugs were considered. RESULTS: Out of 129 patients, 26 (20%) were treated with AC/AP drugs. Warfarin, acenocumarine, LMWH as well as AP were used in 7, 11 and 12 patients, respectively. Adverse events (bleeding) were observed in 3 patients (11.5%), 2 cases being on warfarin (replaced by LMWH) and 1 being AP (suspension without replacement); out of the 3 patients with bleeding, none presented thrombocytopenia. CONCLUSIONS: Despite the frequent findings of hemostatic disorders in a population of frail patients managed in a home care setting, our experience demonstrated that the use of AC/AP drugs has been very rarely responsible for significant complications. PMID- 21633624 TI - Emergency total thyroidectomy for bleeding anaplastic thyroid carcinoma: a viable option for palliation. AB - Anaplastic thyroid carcinoma (ATC) is a rare and highly aggressive thyroid neoplasm. Bleeding from tumor is an uncommon, but potentially life-threatening complication requiring sophisticated intervention facilities which are not usually available at odd hours in emergency. We report the case of a 45-year-old woman who presented with exsanguinating hemorrhage from ATC and was treated by emergency total thyroidectomy. The patient is well three months postoperatively. Emergency total thyroidectomy is a viable option for palliation in ATC presenting with bleeding. PMID- 21633625 TI - Rehabilitation of Patient with Acquired Maxillary Defect, using a Closed Hollow Bulb Obturator. AB - Palliative care means providing support and care for patients with life threatening or debilitating illness so that they can live their life as comfortably as possible. The fact that cure is no longer a reality does not mean that care cannot be made available. Partial maxillectomy defect presents a prosthodontic challenge in terms of re-establishing oronasal separation. Such defect has direct effect on cosmetic, function and psychology of the patient. This article describes step by step clinical and laboratory procedures involved in the rehabilitation of a hemimaxillectomy patient, using a definitive closed hollow bulb obturator, which improved his physical, emotional, functional, social and spiritual needs. PMID- 21633623 TI - Reporting characteristics of cancer pain: a systematic review and quantitative analysis of research publications in palliative care journals. AB - OBJECTIVE: A common disorder requiring symptom palliation in palliative and end of-life care is cancer. Cancer pain is recognized as a global health burden. This paper sought to systematically examine the extent to which there is an adequate scientific research base on cancer pain and its reporting characteristics in the palliative care journal literature. MATERIALS AND METHODS: Search conducted in MEDLINE and CINAHL sought to locate all studies published in 19 palliative/ hospice/ supportive/ end-of-life care journals from 2009 to 2010. The journals included were: American Journal of Hospice and Palliative Care, BMC Palliative Care, Current Opinion in Supportive and Palliative Care, End of Life Care Journal, European Journal of Palliative Care, Hospice Management Advisor, Indian Journal of Palliative Care, International Journal of Palliative Nursing, Internet Journal of Pain Symptom Control and Palliative Care, Journal of Pain and Palliative Care Pharmacotherapy, Journal of Palliative Care, Journal of Palliative Medicine, Journal of Social Work in End-of-life and Palliative Care, Journal of Supportive Oncology, Palliative Medicine, Palliative and Supportive Care, and Supportive Care in Cancer. Journal contents were searched to identify studies that included cancer pain in abstract. RESULTS: During the years 2009 and 2010, of the selected 1,569 articles published in the journals reviewed, only 5.86% (92 articles) were on cancer pain. CONCLUSION: While researchers in the field of palliative care have studied cancer pain, the total percentage for studies is still a low 5.86%. To move the field of palliative care forward so that appropriate guidelines for cancer pain management can be developed, it is critical that more research be reported upon which to base cancer pain therapy in an evidence-based palliative care model. PMID- 21633626 TI - Role of surgery in breast metastasis from carcinoma of the cervix. AB - Carcinoma of the cervix is the most common malignancy among women in India. Although metastatic disease is common, metastasis to breast is rare. A limited number of case reports are published in the world literature. Most of the previous reports of metastatic cervical carcinoma to breast are either autopsy series or widely disseminated disease where no treatment options were available. A rare case of cervical carcinoma presenting as metastasis in breast is reported here where palliative mastectomy improved the general condition of the patient. A female patient aged 58 years was diagnosed and treated for cervical carcinoma, FIGO stage 2B. Four months after the treatment which included both external beam and intracavitory radiotherapy, the patient presented with breast and lung metastasis. Palliative mastectomy was done which improved the general condition of the patient. Metastatic carcinoma of the cervix can present as a case of breast carcinoma. In an appropriate setting, this possibility should be kept in mind. Palliative mastectomy should be offered for patients of cervical carcinoma with metastasis to breast when needed. PMID- 21633627 TI - Palliative care in children: the role of a pediatric dentist in the Indian scenario. PMID- 21633628 TI - Battling Chemotherapy-induced Nausea and Vomiting in Cancer Palliative Care. PMID- 21633629 TI - Mechanism-based Classification of Pain for Physical Therapy Management in Palliative care: A Clinical Commentary. AB - Pain relief is a major goal for palliative care in India so much that most palliative care interventions necessarily begin first with pain relief. Physical therapists play an important role in palliative care and they are regarded as highly proficient members of a multidisciplinary healthcare team towards management of chronic pain. Pain necessarily involves three different levels of classification-based upon pain symptoms, pain mechanisms and pain syndromes. Mechanism-based treatments are most likely to succeed compared to symptomatic treatments or diagnosis-based treatments. The objective of this clinical commentary is to update the physical therapists working in palliative care, on the mechanism-based classification of pain and its interpretation, with available therapeutic evidence for providing optimal patient care using physical therapy. The paper describes the evolution of mechanism-based classification of pain, the five mechanisms (central sensitization, peripheral neuropathic, nociceptive, sympathetically maintained pain and cognitive-affective) are explained with recent evidence for physical therapy treatments for each of the mechanisms. PMID- 21633630 TI - St. Gallen 2011: Summary of the Consensus Discussion. AB - SUMMARY: The 2011 St. Gallen Consensus Conference on early breast cancer provided mostly evidence-based treatment recommendations with a broad spectrum of acceptable clinical practice for global breast cancer care. This report summarizes the results of the 2011 international panel voting procedures with regard to locoregional and endocrine treatment, chemotherapy, targeted therapy as well as adjuvant bisphosphonate use. PMID- 21633631 TI - Study of vascular injuries using endothelial denudation model and the therapeutic application of shock wave: a review. AB - As death toll from cardiovascular diseases has reached historic heights in the developed world, research efforts have been focused on both the understanding of disease progression and also the choice of appropriate treatment strategies. Moreover, to facilitate research, an appropriate animal model is needed to mimic the pathological changes and follow treatment results. This article reviewed the disease mechanisms underlying vascular injuries and also the animal model of endothelial denudation using balloon catheter. On the other hand, the biological effects of shock wave including angiogenesis and the suppression of inflammation were reviewed. Its therapeutic impact on the cardiovascular system and its potential clinical application as well as limitations were also discussed. PMID- 21633632 TI - Rabbit monoclonal antibody: potential application in cancer therapy. AB - By targeting antigens specifically, monoclonal antibodies represent a new class of therapeutic agents for the clinical management of various diseases including cancers. Monoclonal antibody technology has been greatly developed by reducing murine content in antibodies to minimize side effects in clinical applications. However, several intrinsic disadvantages of antibodies with murine origin limit the clinical efficacy of monoclonal antibodies based targeted therapy. The development of rabbit monoclonal antibody technology provides an alternative source of monoclonal antibodies with higher specificity and less cost for the development of routine targeted therapy against cancers. PMID- 21633633 TI - Phase I and Phase II clinical trials of androst-5-ene-3beta,7beta,17beta-triol. AB - The immune regulating DHEA metabolite, androst-5-ene-3beta,7beta,17beta-triol (betaAET), was evaluated for safety, cholesterol lowering, and vaccine enhancement in phase I and phase II clinical trials. Safety and pharmacokinetics were evaluated in one study of normal subjects that received betaAET or placebo transmucosally (buccal tablets) for 4 days. In a second study betaAET was given by daily subcutaneous injection for 3 days. betaAET was subsequently evaluated in placebo-controlled trials for cholesterol lowering in hyperlipidemic subjects and for potentiation of hepatitis B surface antigen (HBsAg) vaccine in elderly subjects. Adverse events were primarily associated with injection site reactions. Pharmacokinetics indicated that betaAET was rapidly cleared after either route of administration in both normal and elderly subjects. Plasma betaAET concentrations typically declined below the limit of detection within a few hours of administration. betaAET pharmacokinetics was similar in males and females and in normal and elderly subjects. betaAET significantly lowered cholesterol in normal adult, but not in elderly or hyperlipidemic subjects. HBsAg titers were not increased in elderly betaAET treated subjects relative to placebo. CONCLUSIONS: Short-term administration of betaAET is safe in humans. betaAET has a cholesterol lowering effect in healthy humans, but not hyperlipidemics. Exogenous betaAET appeared to be rapidly metabolized, which may be consequential to the lack of pharmacological activity. A longer duration of betaAET treatment with higher doses or chemical derivatives that are resistant to metabolic inactivation are likely necessary to treat human disease. The utility of betaAET in humans may be limited to maintenance of homeostasis in healthy adults. PMID- 21633634 TI - Targeting voltage-gated sodium channels for treatment for chronic visceral pain. AB - Voltage-gated sodium channels (VGSCs) play a fundamental role in controlling cellular excitability, and their abnormal activity is related to several pathological processes, including cardiac arrhythmias, epilepsy, neurodegenerative diseases, spasticity and chronic pain. In particular, chronic visceral pain, the central symptom of functional gastrointestinal disorders such as irritable bowel syndrome, is a serious clinical problem that affects a high percentage of the world population. In spite of intense research efforts and after the dedicated decade of pain control and research, there are not many options to treat chronic pain conditions. However, there is a wealth of evidence emerging to give hope that a more refined approach may be achievable. By using electronic databases, available data on structural and functional properties of VGSCs in chronic pain, particularly functional gastrointestinal hypersensitivity, were reviewed. We summarize the involvement and molecular bases of action of VGSCs in the pathophysiology of several organic and functional gastrointestinal disorders. We also describe the efficacy of VGSC blockers in the treatment of these neurological diseases, and outline future developments that may extend the therapeutic use of compounds that target VGSCs. Overall, clinical and experimental data indicate that isoform-specific blockers of these channels or targeting of their modulators may provide effective and novel approaches for visceral pain therapy. PMID- 21633635 TI - Review of screening for pancreatic cancer in high risk individuals. AB - Pancreatic cancer is difficult to diagnose at an early stage and is associated with a very poor survival. Ten percent of pancreatic cancers result from genetic susceptibility and/or familial aggregation. Individuals from families with multiple affected first-degree relatives and those with a known cancer-causing genetic mutation have been shown to be at much higher risk of developing pancreatic cancer. Recent efforts have focused on detecting disease at an earlier stage to improve survival in these high-risk groups. This article reviews high risk groups, screening methods, and current screening programs and their results. PMID- 21633636 TI - Surgicopathological classification of hepatic space-occupying lesions: a single center experience with literature review. AB - Accompanying rapid developments in hepatic surgery, the number of surgeries and identifications of histological types of primary hepatic space-occupying lesions (PHSOLs) have increased dramatically. This has led to many changes in the surgicopathological spectrum of PHSOLs, and has contributed to a theoretical basis for modern hepatic surgery and oncological pathology. Between 1982 and 2009 at the Eastern Hepatobiliary Surgery Hospital (EHBH) in Shanghai, 31 901 patients underwent surgery and were diagnosed as having a PHSOL. In this paper, we present an analysis of the PHSOL cases at the EHBH for this time period, along with results from a systematic literature review. We describe a surgicopathological spectrum comprising more than 100 types of PHSOLs that can be stratified into three types: tumor-like, benign, and malignant. We also stratified the PHSOLs into six subtypes derived from hepatocytes; cholangiocytes; vascular, lymphoid and hemopoietic tissues; muscular, fibrous and adipose tissues; neural and neuroendocrine tissues; and miscellaneous tissues. The present study provides a new classification system that can be used as a current reference for clinicians and pathologists to make correct diagnoses and differential diagnoses among various PHSOLs. PMID- 21633637 TI - Inhibitory effect of schisandrin B on free fatty acid-induced steatosis in L-02 cells. AB - AIM: To investigate the effects of schisandrin B (Sch B) on free fatty acid (FFA) induced steatosis in L-02 cells. METHODS: Cellular steatosis was induced by incubating L-02 cells with a FFA mixture (oleate and palmitate at the ratio of 2:1) for 24 h. Cytotoxicity and apoptosis were evaluated by 3-(4, 5 dmethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assay and Annexin V/propidium iodide staining, respectively. Cellular total lipid was determined using a photocolorimetric method after Nile red staining, and triglyceride content was measured using an enzymatic kit. To study the effects of Sch B on steatosis, L-02 cells were treated with Sch B (1-100 MUmol/L) in the absence or presence of 1 mmol/L FFA for 24 h, and cellular total lipid and triglyceride levels were measured. To explore the mechanisms of action of Sch B in the steatotic L-02 cells, mRNA levels of several regulators of hepatic lipid metabolism including adipose differentiation related protein (ADRP), sterol regulatory element binding protein 1 (SREBP-1), peroxisome proliferator-activated receptor (PPAR)-alpha and PPAR-gamma were measured by quantitative real-time polymerase chain reaction (PCR), and protein levels of ADRP and SREBP-1 were measured by immunoblotting. RESULTS: Treatment with 1 mmol/L FFA for 24 h induced intracellular lipid accumulation in L-02 cells comparable to that in human steatotic livers without causing apparent apoptosis and cytotoxicity. Sch B mitigated cellular total lipid and triglyceride accumulations in the steatotic L 02 cells in a dose-dependent manner. Quantitative real-time PCR and Western blot analyses revealed that treatment of L-02 cells with 100 MUmol/L Sch B reverted the FFA-stimulated up-regulation of ADRP and SREBP-1. CONCLUSION: Sch B inhibits FFA-induced steatosis in L-02 cells by, at least in part, reversing the up regulation of ADRP and SREBP-1. PMID- 21633638 TI - CXCR4/SDF-1 axis is involved in lymph node metastasis of gastric carcinoma. AB - AIM: To investigate the role of CXC chemokine receptor-4 (CXCR4) and stromal cell derived factor-1 (SDF-1) in lymph node metastasis of gastric carcinoma. METHODS: In 40 cases of gastric cancer, expression of CXCR4 mRNA in cancer and normal mucous membrane and SDF-1 mRNA in lymph nodes around the stomach was detected using quantitative polymerase chain reaction (PCR) (TaqMan) and immunohistochemistry assay. SGC-7901 and MGC80-3 cancer cells were used to investigate the effect of SDF-1 on cell proliferation and migration. RESULTS: Quantitative reverse transcription PCR and immunohistochemistry revealed that the expression level of CXCR4 in gastric cancer was significantly higher than that in normal mucous membrane (1.6244 +/- 1.3801 vs 1.0715 +/- 0.5243, P < 0.05). The expression level of CXCR4 mRNA in gastric cancer with lymph node metastasis was also significantly higher than that without lymph node metastasis (0.823 +/- 0.551 vs 0.392 +/- 0.338, P < 0.05). CXCR4 expression was significantly related to poorly differentiated, high tumor stage and lymph node metastasis. Significant differences in the expression level of SDF-1 mRNA were found between lymph nodes in metastatic gastric cancer and normal nodes (0.5432 +/- 0.4907 vs 0.2640 +/- 0.2601, P < 0.05). The positive expression of SDF-1 mRNA in lymph nodes of metastatic gastric cancer was consistent with the positive expression of CXCR4 mRNA in gastric cancer (r = 0.776, P < 0.01). Additionally, human gastric cancer cell lines expressed CXCR4 and showed vigorous proliferation and migratory responses to SDF-1. AMD3100 (a specific CXCR4 antagonist) was also found to effectively reduce the migration of gastric cancer cells. CONCLUSION: The CXCR4/SDF-1 axis is involved in the lymph node metastasis of gastric cancer. CXCR4 is considered as a potential therapeutic target in the treatment of gastric cancer. PMID- 21633639 TI - Protective effects of ACLF sera on metabolic functions and proliferation of hepatocytes co-cultured with bone marrow MSCs in vitro. AB - AIM: To investigate whether the function of hepatocytes co-cultured with bone marrow mesenchymal stem cells (MSCs) could be maintained in serum from acute-on chronic liver failure (ACLF) patients. METHODS: Hepatocyte supportive functions and cytotoxicity of sera from 18 patients with viral hepatitis B-induced ACLF and 18 healthy volunteers were evaluated for porcine hepatocytes co-cultured with MSCs and hepatocyte mono-layered culture, respectively. Chemokine profile was also examined for the normal serum and liver failure serum. RESULTS: Hepatocyte growth factor (HGF) and Tumor necrosis factor; tumor necrosis factor (TNF)-alpha were remarkably elevated in response to ACLF while epidermal growth factor (EGF) and VEGF levels were significantly decreased. Liver failure serum samples induced a higher detachment rate, lower viability and decreased liver support functions in the homo-hepatocyte culture. Hepatocytes co-cultured with MSCs could tolerate the cytotoxicity of the serum from ACLF patients and had similar liver support functions compared with the hepatocytes cultured with healthy human serum in vitro. In addition, co-cultured hepatocytes maintained a proliferative capability despite of the insult from liver failure serum. CONCLUSION: ACLF serum does not impair the cell morphology, viability, proliferation and overall metabolic capacities of hepatocyte co-cultured with MSCs in vitro. PMID- 21633640 TI - Incidence of brain metastasis in patients with esophageal carcinoma. AB - AIM: To determine the incidence of brain metastasis in a contemporary group of patients with carcinoma of the esophagus. METHODS: Retrospective analysis of 53 patients with esophageal carcinoma who received radiotherapy as a component of treatment between 1998 and 2007, including patient and tumor characteristics, and subsequent diagnosis of brain metastasis. The association between the histological type of esophageal cancer and the incidence of brain metastasis was assessed using Fisher's exact test. RESULTS: Forty-four of the fifty-three patients in this study had adenocarcinoma and nine had squamous cell carcinoma, ranging from stage IIA-IVB. Primary treatment was surgery with neoadjuvant chemoradiotherapy (trimodality therapy) in 19% of patients; chemoradiotherapy in 42%; and surgery and adjuvant radiotherapy in 7%. Twenty-five percent of patients in this study received palliative radiotherapy. The overall incidence of brain metastasis in this cohort was 13%. Adenocarcinoma was the primary tumor histology in all of the patients who developed brain metastasis, representing an incidence of 16% in this subgroup. No patients with squamous cell carcinoma received trimodality therapy. The association between histology and brain metastasis was not statistically significant. CONCLUSION: The incidence of brain metastasis in this contemporary cohort of patients with esophageal carcinoma is higher than previously reported and was confined to those with adenocarcinoma. PMID- 21633641 TI - Resected specimen evaluation, anorectal manometry, endoanal ultrasonography and clinical follow-up after STARR procedures. AB - AIM: To investigate stapled transanal rectal resection (STARR) procedures as surgical techniques for obstructed defecation syndrome (ODS) by analyzing specimen evaluation, anorectal manometry, endoanal ultrasonography and clinical follow-up. METHODS: From January to December 2007, we have treated 30 patients. Fifteen treated with double PPH-01 staplers and 15 treated using new CCS 30 contour. Resected specimen were measured with respect to average surface and volume. All patients have been evaluated at 24 mo with clinical examination, anorectal manometry and endoanal ultrasonography. RESULTS: Average surface in the CCS 30 group was 54.5 cm2 statistically different when compared to the STARR group (36.92 cm2). The average volume in the CCS 30 group was 29.8 cc, while in the PPH-01 it was 23.8 cc and difference was statistically significant. The mean hospital stay in the CCS 30 group was 3.1 d, while in the PPH-01 group the median hospital stay was 3.4 d. As regards the long-term follow-up, an overall satisfactory rate of 83.3% (25/30) was achieved. Endoanal ultrasonography performed 1 year following surgery was considered normal in both of the studied groups. Mean resting pressure was higher than the preoperative value (67.2 mmHg in the STARR group and 65.7 mmHg in the CCS30 group vs 54.7 mmHg and 55.3 mmHg, respectively). Resting and squeezing pressures were lower in those patients not satisfied, but data are not statistically significant. CONCLUSION: The STARR procedure with two PPH-01 is a safe surgical procedure to correct ODS. The new Contour CCS 30 could help to increase the amount of the resected tissue without differences in early complications, post-operative pain and in hospital stay compared to the STARR with two PPH-01 technique. PMID- 21633642 TI - p53 antibodies, metallothioneins, and oxidative stress markers in chronic ulcerative colitis with dysplasia. AB - AIM: To investigate the role of p53 antibodies (p53Abs), metallothioneins (MTs) and oxidative stress markers in the early detection of dysplasia in chronic ulcerative colitis (UC). METHODS: The study included 30 UC patients, 15 without dysplasia (group II) and 15 with dysplasia (group III), in addition to 15 healthy volunteers (group I, control subjects). The enzyme-linked immunosorbent assay technique was used to measure serum p53Abs and MTs, while advanced oxidation protein products (AOPPs), and reduced glutathione (GSH) levels were measured by spectrophotometric method in all subjects. RESULTS: In group II and group III compared to group I, there were significant increases in serum levels of AOPPs (145.94 +/- 29.86 MUmol/L and 192.21 +/- 46.71 MUmol/L vs 128.95 +/- 3.06 MUmol/L, P < 0.002 and P < 0.001, respectively), MTs (8.18 +/- 0.35 MUg/mL and 9.20 +/- 0.58 MUg/mL vs 6.12 +/- 0.25 MUg/mL, P < 0.05 and P < 0.05, respectively), and p53Abs (20.19 +/- 3.20 U/mL and 34.66 +/- 1.34 U/mL vs 9.42 +/ 1.64 U/mL, P < 0.001 and P < 0.001, respectively). There were significantly higher levels of AOPPs (P < 0.05) and p53Abs (P < 0.001) in UC patients with dysplasia compared to those without dysplasia, while MTs showed no significant difference between the 2 groups (P > 0.096). In contrast, GSH levels showed a significant decrease in both patients' groups (1.87 +/- 0.02 MUmol/mL and 1.37 +/ 0.09 MUmol/mL vs 2.49 +/- 0.10 MUmol/mL, P < 0.05 and P < 0.05 in groups II and III, respectively) compared with group I, and the levels were significantly lower in group III than group II (P < 0.05). There was a positive correlation between AOPPs and both MTs (r = 0.678, P < 0.001) and p53Abs (r = 0.547, P < 0.001), and also between p53Abs and MTs (r = 0.739, P < 0.001). There was a negative correlation between AOPPs and GSH (r = -0.385, P < 0.001), and also between GSH and both MTs (r = -0.662, P < 0.001) and p53Abs (r = -0.923, P < 0.001). CONCLUSION: Oxidative stress and oxidative cellular damage play an important role in the pathogenesis of chronic UC and the associated carcinogenetic process. p53Abs levels could help in early detection of dysplasia in these conditions. PMID- 21633643 TI - Monoclonal immunoscintigraphy for detection of metastasis and recurrence of colorectal cancer. AB - AIM: To assess the clinical role of monoclonal immunoscintigraphy for the detection of metastasis and recurrence of colorectal cancer. METHODS: Monoclonal immunoscintigraphy was performed in patients operated on for colorectal adenocarcinoma suspected of local recurrence and metastatic disease. The results were compared with conventional diagnostics. RESULTS: Immunoscintigraphic investigation was done in 53 patients. Tumor recurrence occurred in 38 patients, and was confirmed by other diagnostic modalities in 35. In 15 patients, immunoscintigraphic findings were negative, and confirmed in 14 with other diagnostic methods. Comparative analysis confirmed good correlation of immunoscintigraphic findings and the results of conventional diagnostics and the level of tumor marker carcinoembryonic antigen. Statistical analysis of parameters of radiopharmaceutical groups imacis, indimacis and oncoscint presented homogenous characteristics all of three radiopharmaceuticals. The analysis of immunoscintigraphic target focus was clearly improved using tomography. CONCLUSION: Immunoscintigraphy is highly specific and has a good predictive value in local recurrence of colorectal cancer. PMID- 21633644 TI - Long-term outcome and efficacy of endoscopic hemorrhoid ligation for symptomatic internal hemorrhoids. AB - AIM: To assess the long-term outcome of endoscopic hemorrhoid ligation (EHL) for the treatment of symptomatic internal hemorrhoids. METHODS: A total of 759 consecutive patients (415 males and 344 females) were enrolled. Clinical presentations were rectal bleeding (593 patients) and mucosal prolapse (166 patients). All patients received EHL at outpatient clinics. Hemorrhoid severity was classified by Goligher's grading. The mean follow-up period was 55.4 mo (range, 45-92 mo). RESULTS: The number of band ligations averaged 2.35 in the first session for bleeding and 2.69 for prolapsed patients. Bleeding was controlled in 587 (98.0%) patients, while prolapse was reduced in 137 (82.5%) patients. After treatment, 93 patients experienced anal pain and 48 patients had mild bleeding. Patient subjective satisfaction was 93.6%. Repeat treatment or surgery was performed if symptoms were not relieved in the first session. In the bleeding group, the recurrence rate was 3.7% (22 patients) at 1 year, and 6.6% and 13.0% at 2 and 5 years. In the prolapsed group, the recurrence rate was 3.0%, 9.6% and 16.9% at 1, 2 and 5 years, respectively. CONCLUSION: EHL is an easy and well-tolerated procedure for the treatment of symptomatic internal hemorrhoids, with good long-term results. PMID- 21633645 TI - Effects of appendectomy and oral tolerance on dextran sulfate sodium colitis. AB - AIM: To evaluate the concomitant effects of appendectomy and oral tolerance on colitis. METHODS: Delayed-type hypersensitivity (DTH) was investigated at a 7-d interval after ovalbumin (OVA) administration and immunization under normal and colitis conditions in appendectomized or sham-operated mice. Pathological scores for the colon were graded after ingestion of colon-extracted protein (CEP) and induction of dextran sulfate sodium (DSS) colitis in appendectomized or sham operated mice. Thereafter, Th1 and Th2 in Peyer's patches and spleen lymphocytes were detected in CEP-treated and bovine serum albumin (BSA)-treated control mice. RESULTS: In appendectomized mice, DTH was not inhibited at day 7 after OVA administration and at the initial phase of DSS colitis, whereas it was inhibited at day 14 and day 21. However, in sham-operated mice, it was inhibited during the whole procedure and the onset of DSS colitis. The protective role of CEP against DSS colitis was present in sham-operated mice, with predominant improvement of colonic pathological changes, while vanished in the appendectomized mice. A shift from Th1 to Th2 in Peyer's patches resulted from a decrease of Th1 cells with the ingestion of CEP. Compared with BSA in the sham-operated group, no predominant changes were observed in the appendectomized mice. CONCLUSION: Appendectomy interferes with the protective role of CEP in DSS colitis via a shift from Th2 to Th1 during oral tolerance induction. PMID- 21633646 TI - A case of steroid-dependent myeloid granulocytic sarcoma masquerading as Crohn's disease. AB - Small bowel tumors and Crohn's disease are common causes of small bowel obstruction. Early stage neoplasms can easily be mistaken for Crohn's disease. Therefore, thorough work-ups including imaging studies and endoscopic evaluation with biopsies are critical for accurate diagnosis. Here we report a case of an otherwise healthy female with progressive onset of multiple, recurrent obstructive symptoms secondary to terminal ileal narrowing who was referred for management of steroid-dependent Crohn's disease. After thorough evaluation, the diagnosis was revised to myeloid granulocytic sarcoma involving the terminal ileum. In this case, a delay in diagnosis can be detrimental for prognosis, as myeloid granulocytic sarcoma is highly predictive of underlying acute myeloid leukemia and needs urgent referral for chemotherapy and/or resection. PMID- 21633647 TI - Optimized management of advanced hepatocellular carcinoma: four long-lasting responses to sorafenib. AB - The therapeutic options for hepatocellular carcinoma (HCC) have been so far rather inadequate. Sorafenib has shown an overall survival benefit and has become the new standard of care for advanced HCC. Nevertheless, in clinical practice, some patients are discontinuing this drug because of side effects, and misinterpretation of radiographic response may contribute to this. We highlight the importance of prolonged sorafenib administration, even at reduced dose, and of qualitative and careful radiographic evaluation. We observed two partial and two complete responses, one histologically confirmed, with progression-free survival ranging from 12 to 62 mo. Three of the responses were achieved following substantial dose reductions, and a gradual change in lesion density preceded or paralleled tumor shrinkage, as seen by computed tomography. This report supports the feasibility of dose adjustments to allow prolonged administration of sorafenib, and highlights the need for new imaging criteria for a more appropriate characterization of response in HCC. PMID- 21633648 TI - Severe alcoholic hepatitis: glucocorticoid saves lives and transplantation is promising. AB - Glucocorticosteroids have been used as the only treatment for a long time which significantly reduced the mortality of the patients with severe alcoholic hepatitis. The efficacy of transplantation has been recently addressed in a pilot study. The result seems promising but needs larger multicenter trials. PMID- 21633649 TI - An annual topic highlight: alcohol and liver, 2011. AB - AN ANNUAL TOPIC HIGHLIGHT: Alcohol and Liver, 2011, covers the important and new aspects of pathogenesis of alcoholic liver diseases (ALD). It includes broad topics ranging from the exacerbation of ALD by infectious (viral) agents (hepatitis C virus and human immunodeficiency virus) to the influence of alcohol on liver fibrogenesis, lipid rafts, autophagy and other aspects. This issue is recommended for both basic scientists and clinicians who are involved in alcoholic liver research. PMID- 21633650 TI - Epigenetic regulation in alcoholic liver disease. AB - Alcoholic liver disease (ALD) is characterized by steatosis or fat deposition in the liver and inflammation, which leads to cirrhosis and hepatocellular carcinoma. Induction of target genes without involving changes in DNA sequence seems to contribute greatly to liver injury. Chromatin modifications including alterations in histones and DNA, as well as post-transcriptional changes collectively referred to as epigenetic effects are altered by alcohol. Recent studies have pointed to a significant role for epigenetic mechanisms at the nucleosomal level influencing gene expression and disease outcome in ALD. Specifically, epigenetic alterations by alcohol include histone modifications such as changes in acetylation and phosphorylation, hypomethylation of DNA, and alterations in miRNAs. These modifications can be induced by alcohol-induced oxidative stress that results in altered recruitment of transcriptional machinery and abnormal gene expression. Delineating these mechanisms in initiation and progression of ALD is becoming a major area of interest. This review summarizes key epigenetic mechanisms that are dysregulated by alcohol in the liver. Alterations by alcohol in histone and DNA modifications, enzymes related to histone acetylation such as histone acetyltransferases, histone deacetylases and sirtuins, and methylation enzymes such as DNA methyltransferases are discussed. Chromatin modifications and miRNA alterations that result in immune cell dysfunction contributing to inflammatory cytokine production in ALD is reviewed. Finally, the role of alcohol-mediated oxidative stress in epigenetic regulation in ALD is described. A better understanding of these mechanisms is crucial for designing novel epigenetic based therapies to ameliorate ALD. PMID- 21633651 TI - Histone modifications and alcohol-induced liver disease: are altered nutrients the missing link? AB - Alcoholism is a major health problem in the United States and worldwide, and alcohol remains the single most significant cause of liver-related diseases and deaths. Alcohol is known to influence nutritional status at many levels including nutrient intake, absorption, utilization, and excretion, and can lead to many nutritional disturbances and deficiencies. Nutrients can dramatically affect gene expression and alcohol-induced nutrient imbalance may be a major contributor to pathogenic gene expression in alcohol-induced liver disease (ALD). There is growing interest regarding epigenetic changes, including histone modifications that regulate gene expression during disease pathogenesis. Notably, modifications of core histones in the nucleosome regulate chromatin structure and DNA methylation, and control gene transcription. This review highlights the role of nutrient disturbances brought about during alcohol metabolism and their impact on epigenetic histone modifications that may contribute to ALD. The review is focused on four critical metabolites, namely, acetate, S-adenosylmethionine, nicotinamide adenine dinucleotide and zinc that are particularly relevant to alcohol metabolism and ALD. PMID- 21633652 TI - Targeting collagen expression in alcoholic liver disease. AB - Alcoholic liver disease (ALD) is a leading cause of liver disease and liver related deaths globally, particularly in developed nations. Liver fibrosis is a consequence of ALD and other chronic liver insults, which can progress to cirrhosis and hepatocellular carcinoma if left untreated. Liver fibrosis is characterized by accumulation of excess extracellular matrix components, including type I collagen, which disrupts liver microcirculation and leads to injury. To date, there is no therapy for the treatment of liver fibrosis; thus treatments that either prevent the accumulation of type I collagen or hasten its degradation are desirable. The focus of this review is to examine the regulation of type I collagen in fibrogenic cells of the liver and to discuss current advances in therapeutics to eliminate excessive collagen deposition. PMID- 21633653 TI - Fibronectin: functional character and role in alcoholic liver disease. AB - Fibronectins are adhesive glycoproteins that can be found in tissue matrices and circulating in various fluids of the body. The variable composition of fibronectin molecules facilitates a diversity of interactions with cell surface receptors that suggest a role for these proteins beyond the structural considerations of the extracellular matrix. These interactions implicate fibronectin in the regulation of mechanisms that also determine cell behavior and activity. The two major forms, plasma fibronectin (pFn) and cellular fibronectin (cFn), exist as balanced amounts under normal physiological conditions. However, during injury and/or disease, tissue and circulating levels of cFn become disproportionately elevated. The accumulating cFn, in addition to being a consequence of prolonged tissue damage, may in fact stimulate cellular events that promote further damage. In this review, we summarize what is known regarding such interactions between fibronectin and cells that may influence the biological response to injury. We elaborate on the effects of cFn in the liver, specifically under a condition of chronic alcohol-induced injury. Studies have revealed that chronic alcohol consumption stimulates excess production of cFn by sinusoidal endothelial cells and hepatic stellate cells while impairing its clearance by other cell types resulting in the build up of this glycoprotein throughout the liver and its consequent increased availability to influence cellular activity that could promote the development of alcoholic liver disease. We describe recent findings by our laboratory that support a plausible role for cFn in the promotion of liver injury under a condition of chronic alcohol abuse and the implications of cFn stimulation on the pathogenesis of alcoholic liver disease. These findings suggest an effect of cFn in regulating cell behavior in the alcohol-injured liver that is worth further characterizing not only to gain a more comprehensive understanding of the role this reactive glycoprotein plays in the progression of injury but also for the insight further studies could provide towards the development of novel therapies for alcoholic liver disease. PMID- 21633654 TI - Mechanisms of alcohol-mediated hepatotoxicity in human-immunodeficiency-virus infected patients. AB - Clinical observations have demonstrated that excessive chronic alcohol use negatively affects human immunodeficiency virus (HIV) infection and contributes to the liver manifestations of the disease, even in HIV mono-infection. HIV/hepatitis C virus (HCV) co-infection is associated with increased progression of HVC liver disease compared to HCV infection alone, and both of these are negatively affected by alcohol use. Recent data suggest that alcohol use and HIV infection have common targets that contribute to progression of liver disease. Both HIV infection and chronic alcohol use are associated with increased gut permeability and elevated plasma levels of lipopolysaccharide; a central activator of inflammatory responses. Both alcoholic liver disease and HIV infection result in non-specific activation of innate immunity, proinflammatory cytokine cascade upregulation, as well as impaired antigen presenting cell and dendritic cell functions. Finally, alcohol, HIV and antiretroviral therapy affect hepatocyte functions, which contributes to liver damage. The common targets of alcohol and HIV infection in liver disease are discussed in this mini-review. PMID- 21633655 TI - Involvement of autophagy in alcoholic liver injury and hepatitis C pathogenesis. AB - This review describes the principal pathways of macroautophagy (i.e. autophagy), microautophagy and chaperone-mediated autophagy as they are currently known to occur in mammalian cells. Because of its crucial role as an accessory digestive organ, the liver has a particularly robust autophagic activity that is sensitive to changes in plasma and dietary components. Ethanol consumption causes major changes in hepatic protein and lipid metabolism and both are regulated by autophagy, which is significantly affected by hepatic ethanol metabolism. Ethanol exposure enhances autophagosome formation in liver cells, but suppresses lysosome function. Excessive ethanol consumption synergizes with hepatitis C virus (HCV) to exacerbate liver injury, as alcohol-consuming HCV patients frequently have a longer course of infection and more severe manifestations of chronic hepatitis than abstinent HCV patients. Alcohol-elicited exacerbation of HCV infection pathogenesis is related to modulation by ethanol metabolism of HCV replication. Additionally, as part of this mechanism, autophagic proteins have been shown to regulate viral (HCV) replication and their intracellular accumulation. Because ethanol induces autophagosome expression, enhanced levels of autophagic proteins may enhance HCV infectivity in liver cells of alcoholics and heavy drinkers. PMID- 21633656 TI - Animal models for studying hepatitis C and alcohol effects on liver. AB - Chronic consumption of ethanol has a dramatic effect on the clinical outcome of patients with hepatitis C virus (HCV) infection, but the mechanism linking these two pathologies is unknown. Presently, in vitro systems are limited in their ability to study the interaction between a productive wild-type HCV infection and chronic ethanol exposure. Mouse models are potentially very useful in dissecting elements of the HCV-ethanol relationship. Experiments in mice that transgenically express HCV proteins are outlined, as are experiments for the generation of mice with chimeric human livers. The latter models appear to have the most promise for accurately modeling the effects of chronic ethanol intake in HCV-infected human livers. PMID- 21633658 TI - microRNAs: fad or future of liver disease. AB - microRNAs (miRs) are small non-coding RNAs that regulate both mRNA and protein expression of target genes, which results in alterations in mRNA stability or translation inhibition. miRs influence at least one third of all human transcripts and are known regulators of various important cellular growth and differentiation factors. miRs have recently emerged as key regulatory molecules in chronic liver disease. This review details recent contributions to the field of miRs that influence liver development and the broad spectrum of disease, from non-alcoholic fatty liver disease to fibrosis/cirrhosis, with particular emphasis on hepatic stellate cells and potential use of miRs as therapeutic tools. PMID- 21633657 TI - Role of lipid rafts in liver health and disease. AB - Liver diseases are an increasingly common cause of morbidity and mortality; new approaches for investigation of mechanisms of liver diseases and identification of therapeutic targets are emergent. Lipid rafts (LRs) are specialized domains of cellular membranes that are enriched in saturated lipids; they are small, mobile, and are key components of cellular architecture, protein partition to cellular membranes, and signaling events. LRs have been identified in the membranes of all liver cells, parenchymal and non-parenchymal; more importantly, LRs are active participants in multiple physiological and pathological conditions in individual types of liver cells. This article aims to review experimental-based evidence with regard to LRs in the liver, from the perspective of the liver as a whole organ composed of a multitude of cell types. We have gathered up-to-date information related to the role of LRs in individual types of liver cells, in liver health and diseases, and identified the possibilities of LR-dependent therapeutic targets in liver diseases. PMID- 21633659 TI - Hepatic stellate cells and innate immunity in alcoholic liver disease. AB - Constant alcohol consumption is a major cause of chronic liver disease, and there has been a growing concern regarding the increased mortality rates worldwide. Alcoholic liver diseases (ALDs) range from mild to more severe conditions, such as steatosis, steatohepatitis, fibrosis, cirrhosis, and hepatocellular carcinoma. The liver is enriched with innate immune cells (e.g. natural killer cells and Kupffer cells) and hepatic stellate cells (HSCs), and interestingly, emerging evidence suggests that innate immunity contributes to the development of ALDs (e.g. steatohepatitis and liver fibrosis). Indeed, HSCs play a crucial role in alcoholic steatosis via production of endocannabinoid and retinol metabolites. This review describes the roles of the innate immunity and HSCs in the pathogenesis of ALDs, and suggests therapeutic targets and strategies to assist in the reduction of ALD. PMID- 21633660 TI - Role of MGST1 in reactive intermediate-induced injury. AB - Microsomal glutathione transferase (MGST1, EC 2.5.1.18) is a membrane bound glutathione transferase extensively studied for its ability to detoxify reactive intermediates, including metabolic electrophile intermediates and lipophilic hydroperoxides through its glutathione dependent transferase and peroxidase activities. It is expressed in high amounts in the liver, located both in the endoplasmic reticulum and the inner and outer mitochondrial membranes. This enzyme is activated by oxidative stress. Binding of GSH and modification of cysteine 49 (the oxidative stress sensor) has been shown to increase activation and induce conformational changes in the enzyme. These changes have either been shown to enhance the protective effect ascribed to this enzyme or have been shown to contribute to cell death through mitochondrial permeability transition pore formation. The purpose of this review is to elucidate how one enzyme found in two places in the cell subjected to the same conditions of oxidative stress could both help protect against and contribute to reactive oxygen species-induced liver injury. PMID- 21633661 TI - Proteasome inhibitor treatment in alcoholic liver disease. AB - Oxidative stress, generated by chronic ethanol consumption, is a major cause of hepatotoxicity and liver injury. Increased production of oxygen-derived free radicals due to ethanol metabolism by CYP2E1 is principally located in the cytoplasm and in the mitochondria, which does not only injure liver cells, but also other vital organs, such as the heart and the brain. Therefore, there is a need for better treatment to enhance the antioxidant response elements. To date, there is no established treatment to attenuate high levels of oxidative stress in the liver of alcoholic patients. To block this oxidative stress, proteasome inhibitor treatment has been found to significantly enhance the antioxidant response elements of hepatocytes exposed to ethanol. Recent studies have shown in an experimental model of alcoholic liver disease that proteasome inhibitor treatment at low dose has cytoprotective effects against ethanol-induced oxidative stress and liver steatosis. The beneficial effects of proteasome inhibitor treatment against oxidative stress occurred because antioxidant response elements (glutathione peroxidase 2, superoxide dismutase 2, glutathione synthetase, glutathione reductase, and GCLC) were up-regulated when rats fed alcohol were treated with a low dose of PS-341 (Bortezomib, Velcade((r))). This is an important finding because proteasome inhibitor treatment up-regulated reactive oxygen species removal and glutathione recycling enzymes, while ethanol feeding alone down-regulated these antioxidant elements. For the first time, it was shown that proteasome inhibition by a highly specific and reversible inhibitor is different from the chronic ethanol feeding-induced proteasome inhibition. As previously shown by our group, chronic ethanol feeding causes a complex dysfunction in the ubiquitin proteasome pathway, which affects the proteasome system, as well as the ubiquitination system. The beneficial effects of proteasome inhibitor treatment in alcoholic liver disease are related to proteasome inhibitor reversibility and the rebound of proteasome activity 72 h post PS-341 administration. PMID- 21633662 TI - siRNA targeting Livin decreases tumor in a xenograft model for colon cancer. AB - AIM: To evaluate the effect of silencing Livin gene expression with siRNA to apoptosis and proliferation in a colon cancer cell line. METHODS: To investigate the anticancer effect of silencing Livin gene expression, we established an siRNA transfected cell line using the HCT116 colon cancer cell line. After confirming the successful transfection, MTT assay, flow cytometry and annexin V staining were employed to evaluate the antiapoptotic effect. To confirm the in vivo effect of Livin-siRNA, different doses of Livin-siRNA were injected into xenografted tumors in BALB/c nude mice model. RESULTS: Livin expression was dramatically decreased after siRNA transfection, especially at 25 MUmol/L of siRNA, but this suppression was not dose-dependent. The cell count at 18 h after transfection was significantly reduced as compared with controls (P < 0.01), but tended not to decrease proportionally depending on transfected dose or time. MTT assay revealed that silencing the Livin gene suppressed cellular proliferation at 18 h after transfection (P = 0.04); however, the inhibitory effect disappeared thereafter. Also, there was no significant difference in cellular proliferation depending on siRNA dose. The rate of apoptosis also increased with silencing of the Livin gene. In vivo, the tumor size significantly decreased after Livin-siRNA injection at 20 MUmol/L concentration (P = 0.03). There were no significant body weight changes of mice after siRNA injection. Histologic examination revealed no significant toxic reaction in kidney, liver and brain of mice. CONCLUSION: siRNA mediated downregulation of Livin expression can induce apoptosis in colon cancer in vitro and in vivo, which suggests the possibility of new cancer therapeutics using siRNA. PMID- 21633663 TI - Differential protein expression during colonic adaptation in ultra-short bowel rats. AB - AIM: To investigate the proteins involved in colonic adaptation and molecular mechanisms of colonic adaptation in rats with ultra-short bowel syndrome (USBS). METHODS: Sprague Dawley rats were randomly assigned to three groups: USBS group (10 rats) undergoing an approximately 90%-95% small bowel resection; sham operation group (10 rats) undergoing small bowel transaction and anastomosis; and control group (ten normal rats). Colon morphology and differential protein expression was analyzed after rats were given post-surgical enteral nutrition for 21 d. Protein expression in the colonic mucosa was analyzed by two-dimensional electrophoresis (2-DE) in all groups. Differential protein spots were detected by ImageMaster 2D Platinum software and were further analyzed with matrix-assisted laser desorption/ionization-time-of-flight/time-of-flight-mass spectrometric (MALDI-TOF/TOF-MS) analysis. RESULTS: The colonic mucosal thickness significantly increased in the USBS group compared with the control group (302.1 +/- 16.9 MUm vs 273.7 +/- 16.0 MUm, P < 0.05). There was no statistically significant difference between the sham-operation group and control group (P > 0.05). The height of colon plica markedly improved in USBS group compared with the control group (998.4 +/- 81.2 MUm vs 883.4 +/- 39.0 MUm, P < 0.05). There was no statistically significant difference between the sham-operation and control groups (P > 0.05). A total of 141 differential protein spots were found in the USBS group. Forty-nine of these spots were down-regulated while 92 protein spots were up-regulated by over 2-folds. There were 133 differential protein spots in USBS group. Thirty of these spots were down-regulated and 103 were up-regulated. There were 47 common differential protein spots among the three groups, including 17 down-regulated protein spots and 30 up-regulated spots. Among 47 differential spots, eight up-regulated proteins were identified by MALDI-TOF/TOF-MS. These proteins were previously reported to be involved in sugar and fat metabolism, protein synthesis and oxidation reduction, which are associated with colonic adaption. CONCLUSION: Eight proteins found in this study play important roles in colonic compensation and are associated with sugar and fat metabolism, protein synthesis, and molecular chaperoning. PMID- 21633664 TI - Non-cirrhotic portal hypertension with large regenerative nodules: a diagnostic challenge. AB - Non-cirrhotic portal hypertension is a poorly understood condition characterized by portal hypertension in the absence of conventional hepatic cirrhosis and described in association with blood coagulation disorders, myeloproliferative and immunological diseases and with exposure to toxic drugs. Very recently, precise classification criteria have been proposed in order to define four distinct subcategories. The present case highlights how the clinical presentation, the confounding results from imaging studies, and the difficulties in the histological evaluation often render cases of non-cirrhotic portal hypertension a real diagnostic challenge. It also underscores the classification problems which can be faced once this diagnosis is performed. Indeed, the different subcategories proposed result from the prevalent subtypes in a spectrum of hepatic regenerative responses to a variety of injuries determining microcirculatory disturbances. More flexibility in classification should derive from this etiopathogenic background. PMID- 21633665 TI - The Oxidative State of LDL is the Major Determinant of Anti/Prooxidant Effect of Coffee on Cu Catalysed Peroxidation. AB - Antioxidants exert contrasting effect on low density lipoprotein (LDL) oxidation catalysed by metals, acting as pro-oxidants under select in vitro conditions. Through our study on the effect of coffee on LDL oxidation, we identified the parameters governing this phenomenon, contributing to the comprehension of its mechanism and discovering significant implications for correct alimentary recommendations. By measuring conjugated diene formation, we have analysed the quantitative and qualitative effects exerted by an extract of roasted coffee on LDL oxidation triggered by copper sulphate. When the relative effects of different coffee concentrations were plotted against the lag time (LT) of control LDL (C-LDL), the apparently random experimental data arranged in sensible patterns: by increasing the LT the antioxidant activity of coffee decreased progressively to become prooxidant. The critical LT, at which coffee switches from antioxidant to prooxidant, increased by increasing coffee concentration. Also the contrasting results obtained following a delayed addition of coffee to the assay, arranged in a simple pattern when referred to the LT of C-LDL: the prooxidant effect decreased to become antioxidant as the LT of C-LDL increased. The dependence of coffee effect on the LT of C-LDL was influenced by LDL but not by metal catalyst concentration. These novel findings point to the oxidative state of LDL as a major parameter controlling the anti/prooxidant effect of coffee and suggest the LT of C-LDL as a potent analytical tool to express experimental data when studying the action exerted by a compound on LDL oxidation. PMID- 21633667 TI - Molecular imaging of akt enables early prediction of response to molecular targeted therapy. AB - Development of noninvasive, real-time molecular imaging tools to assess responsiveness of a given therapy may be a critical component of the success of individualized therapy approach for patients. Toward this, we have previously developed and validated molecular sensors for Akt and caspase-3 activity, and in this report, we have explored the utility of these reporters in assessing the responsiveness of tumors to a combination of gemcitabine (Gem) and cetuximab (Cet) delivered in two opposite schedules. We found that human head and neck cancer (UMSCC1) xenografts responded significantly better in a schedule where cetuximab was administered after gemcitabine when compared with the schedule of cetuximab followed by gemcitabine. Wilcoxon two-sample tests suggested that the difference in tumor volumes in two schedules became significant on day 7 (P > .05 on day 4, and P < .05 on days 7 and 10), and the difference in activity of Akt in two schedules became significant on day 4 (P < .05 on days 4, 6, and 10). Using Akt reporter activity and cubic spline interpolation, the distinction between the two schedules could be detected 2 days before using the tumor volume, suggesting that molecular imaging of Akt may allow early prediction of therapy responsiveness. We did not observe a significant difference between the two schedules in the caspase-3 activity. In summary, this proof-of-concept study provides a basis for using molecular imaging of Akt as an early indicator of therapeutic efficacy. PMID- 21633666 TI - Parkin, A Top Level Manager in the Cell's Sanitation Department. AB - Parkin belongs to a class of multiple RING domain proteins designated as RBR (RING, in between RING, RING) proteins. In this review we examine what is known regarding the structure/function relationship of the Parkin protein. Parkin contains three RING domains plus a ubiquitin-like domain and an in-between-RING (IBR) domain. RING domains are rich in cysteine amino acids that act as ligands to bind zinc ions. RING domains may interact with DNA or with other proteins and perform a wide range of functions. Some function as E3 ubiquitin ligases, participating in attachment of ubiquitin chains to signal proteasome degradation; however, ubiquitin may be attached for purposes other than proteasome degradation. It was determined that the C-terminal most RING, RING2, is essential for Parkin to function as an E3 ubiquitin ligase and a number of substrates have been identified. However, Parkin also participates in a number of other fiunctions, such as DNA repair, microtubule stabilization, and formation of aggresomes. Some functions, such as participation in a multi-protein complex implicated in NMDA activity at the post synaptic density, do not require ubiquitination of substrate molecules. Recent observations of RING proteins suggest their function may be regulated by zinc ion binding. We have modeled the three RING domains of Parkin and have identified a new set of RING2 ligands. This set allows for binding of two rather than just one zinc ion, opening the possibility that the number of zinc ions bound acts as a molecular switch to modulate Parkin function. PMID- 21633668 TI - JLK1486, a Bis 8-Hydroxyquinoline-Substituted Benzylamine, Displays Cytostatic Effects in Experimental Gliomas through MyT1 and STAT1 Activation and, to a Lesser Extent, PPARgamma Activation. AB - Gliomas account for 5% to 7% of all solid cancers in adults and up to 30% of solid cancers in children; glioblastomas are the most malignant type of glioma and often have dismal prognoses. The alkylating agent temozolomide provides the greatest chemotherapeutic benefits currently available; however, glioblastoma patients cannot be cured. Novel drugs that efficiently combat glioblastomas are therefore of great interest. We report here that JLK1486, an 8-hydroxyquinoline substituted benzylamine, could represent a novel chemical scaffold to reach this goal. Indeed, JLK1486 mediated anticancer activity in vivo (through intravenous as well as oral routes of administrations) in an orthotopic xenograft model and displayed efficiency similar to that of temozolomide. The therapeutic benefits of JLK1486 seem to relate to its ability to activate various transcription factors (including Myt1, STAT1, and peroxisome proliferator-activated receptor gamma) in glioma cells. These transcription factors are implicated in the control of glioma cell proliferation, and the resultant global effect of their activation by JLK1486 was cytostatic, not cytotoxic. Thus, the current study opens the door for the development of novel compounds to combat glioblastoma using 8 hydroxyquinoline benzylamine analogs. PMID- 21633669 TI - Bioengineered human arginase I with enhanced activity and stability controls hepatocellular and pancreatic carcinoma xenografts. AB - Hepatocellular carcinoma (HCC) and pancreatic carcinoma (PC) cells often have inherent urea cycle defects rendering them auxotrophic for the amino acid l arginine (l-arg). Most HCC and PC require extracellular sources of l-arg and undergo cell cycle arrest and apoptosis when l-arg is restricted. Systemic, enzyme-mediated depletion of l-arg has been investigated in mouse models and human trials. Non-human enzymes elicit neutralizing antibodies, whereas human arginases display poor pharmacological properties in serum. Co(2+) substitution of the Mn(2+) metal cofactor in human arginase I (Co-hArgI) was shown to confer more than 10-fold higher catalytic activity (k(cat)/K(m)) and 5-fold greater stability. We hypothesized that the Co-hArgI enzyme would decrease tumor burden by systemic elimination of l-arg in a murine model. Co-hArgI was conjugated to 5 kDa PEG (Co-hArgI-PEG) to enhance circulation persistence. It was used as monotherapy for HCC and PC in vitro and in vivo murine xenografts. The mechanism of cell death was also investigated. Weekly treatment of 8 mg/kg Co-hArgI-PEG effectively controlled human HepG2 (HCC) and Panc-1 (PC) tumor xenografts (P = .001 and P = .03, respectively). Both cell lines underwent apoptosis in vitro with significant increased expression of activated caspase-3 (P < .001). Furthermore, there was evidence of autophagy in vitro and in vivo. We have demonstrated that Co-hArgI-PEG is effective at controlling two types of l-arg dependent carcinomas. Being a nonessential amino acid, arginine deprivation therapy through Co-hArgI-PEG holds promise as a new therapy in the treatment of HCC and PC. PMID- 21633670 TI - Synergistic Effects of the Green Tea Extract Epigallocatechin-3-gallate and Taxane in Eradication of Malignant Human Prostate Tumors. AB - We have examined whether epigallocatechin-3-gallate (EGCG), and extract of green tea, in combination with taxane (i.e., paclitaxel and docetaxel), exerts a synergistic activity in blocking human prostate PC-3ML tumor cell growth in vitro and in vivo. Growth assays in vitro revealed that the IC(50) values were ~30 uM, ~3 nM, and ~6 nM, for EGCG, paclitaxel and docetaxel, respectively. Isobolograms generated from the data clearly indicated that EGCG in combination with paclitaxel or docetaxel had an additive effect in blocking tumor cell growth. EGCG combined with taxane also had an additive effect to increase the expression of apoptotic genes, (p53, p73, p21, and caspase 3) and the percent apoptosis observed in vitro and in tumor modeling studies in severe combined immunodeficient mice. The tumor modeling studies clearly showed that EGCG plus taxane injected intraperitoneally (i.p.) induced a significant increase in apoptosis rates (TUNEL assays) and eliminated preexisting tumors generated from PC-3ML cells implanted i.p., increasing disease-free survival rates to greater than 90%. More importantly, the combination therapy (i.p. biweekly) blocked metastases after intravenous injection of PC-3ML cells through the tail vein. In mice treated with EGCG plus taxane, the disease-free survival rates increased from 0% (in untreated mice) to more than 70% to 80% in treated mice. Taken together, these data demonstrate for the first time that EGCG in combination with taxane may provide a novel therapeutic treatment of advanced prostate cancer. PMID- 21633672 TI - Feasibility and safety of silicone rubber contrast-enhanced microcomputed tomography in evaluating the angioarchitecture of prostatectomy specimens. AB - This ethics committee-approved pilot study was carried out with informed consent. A protocol was developed to assess the feasibility of in vitro Microfil injection of prostate cancer specimens followed by analysis with micro-computed tomography (microCT) to characterize the functional vascularity of prostatic tissue and evaluate its safety with respect to the preservation of a specimen for pathologic examination. The visible prostatic arteries of two surgically resected prostates frompatients with known prostate cancer (PCa) were injected with MicrofilMV-122 contrast medium immediately after removal. The specimens were scanned using microCT and were qualitatively examined using three-dimensional analysis software (MicroView; GE Healthcare Biosciences). The Microfil perfusion in the two samples was sufficient to view the functional vascularity arising from a major prostatic artery, up to a resolution of 17.626 um without any indication of adverse effects due to Microfil injection. Malignant prostatic regions showed a greater vascular density on histology but decreased vascular perfusion compared with benign prostatic regions. The use of microCT on Microfil-injected prostates seems to be a feasible and specimen-preserving method for visualizing the three-dimensional vessel patterns present in resected human prostates. PMID- 21633671 TI - Molecular Validation of PACE4 as a Target in Prostate Cancer. AB - Prostate cancer remains the single most prevalent cancer in men. Standard therapies are still limited and include androgen ablation that initially causes tumor regression. However, tumor cells eventually relapse and develop into a hormone-refractory prostate cancer. One of the current challenges in this disease is to define new therapeutic targets, which have been virtually unchanged in the past 30 years. Recent studies have suggested that the family of enzymes known as the proprotein convertases (PCs) is involved in various types of cancers and their progression. The present study examined PC expression in prostate cancer and validates one PC, namely PACE4, as a target. The evidence includes the observed high expression of PACE4 in all different clinical stages of human prostate tumor tissues. Gene silencing studies targeting PACE4 in the DU145 prostate cancer cell line produced cells (cell line 4-2) with slower proliferation rates, reduced clonogenic activity, and inability to grow as xenografts in nude mice. Gene expression and proteomic profiling of the 4-2 cell line reveals an increased expression of known cancer-related genes (e.g., GJA1, CD44, IGFBP6) that are downregulated in prostate cancer. Similarly, cancer genes whose expression is decreased in the 4-2 cell line were upregulated in prostate cancer (e.g., MUC1, IL6). The direct role of PACE4 in prostate cancer is most likely through the upregulated processing of growth factors or through the aberrant processing of growth factors leading to sustained cancer progression, suggesting that PACE4 holds a central role in prostate cancer. PMID- 21633673 TI - STAT3 Knockdown in B16 Melanoma by siRNA Lipopolyplexes Induces Bystander Immune Response In Vitro and In Vivo. AB - Persistent activation of STAT3 plays a major role in cancer progression and immune escape. Therefore, targeting STAT3 in tumors is essential to enhance/reactivate antitumor immune response. In our previous studies, we demonstrated the efficacy of stearic acid-modified polyethylenimine (PEI-StA) in promoting small interfering RNA (siRNA) silencing of STAT3 in B16.F10 melanoma in vitro and in vivo. In the current study, we examine the immunologic impact of this intervention. Toward this goal, the infiltration and activation of lymphocytes and dendritic cells (DCs) in the tumor mass were assessed using flow cytometry. Moreover, the levels of IFN-gamma, IL-12, and TNF-alpha in homogenized tumor supernatants were determined. Moreover, mixed lymphocytes reaction using splenocytes of tumor-bearing mice was used to assess DC functionality on siRNA/lipopolyplexes intervention. Our results demonstrated up to an approximately fivefold induction in the infiltration of CD3(+) cells in tumor mass on STAT3 knockdown with high levels of CD4(+), CD8(+), and NKT cells. Consistently, DC infiltration in tumor milieu increased up to approximately fourfold. Those DCs were activated, in an otherwise suppressive microenvironment, as evidenced by a high expression of costimulatory molecules CD86 and CD40. ELISA analysis revealed a significant increase in IFN-gamma, IL-12, and TNF-alpha. Moreover, mixed lymphocytes reaction demonstrated alloreactivity of these DCs as assessed by high T-cell proliferation and IL-2 production. Our results suggest a bystander immune response after local STAT3 silencing by siRNA. This strategy could be beneficial as an adjuvant therapy along with current cancer vaccine formulations. PMID- 21633674 TI - Psychiatric diagnosis: pros and cons of prototypes vs. operational criteria. PMID- 21633675 TI - Lessons learned from radiation disasters. PMID- 21633676 TI - Addressing the mental health consequences of the Japan triple catastrophe. PMID- 21633677 TI - A conceptual framework for the revision of the ICD-10 classification of mental and behavioural disorders. AB - The World Health Organization (WHO) is revising the ICD-10 classification of mental and behavioural disorders, under the leadership of the Department of Mental Health and Substance Abuse and within the framework of the overall revision framework as directed by the World Health Assembly. This article describes WHO's perspective and priorities for mental and behavioural disorders classification in ICD-11, based on the recommendations of the International Advisory Group for the Revision of ICD-10 Mental and Behavioural Disorders. The WHO considers that the classification should be developed in consultation with stakeholders, which include WHO member countries, multidisciplinary health professionals, and users of mental health services and their families. Attention to the cultural framework must be a key element in defining future classification concepts. Uses of the ICD that must be considered include clinical applications, research, teaching and training, health statistics, and public health. The Advisory Group has determined that the current revision represents a particular opportunity to improve the classification's clinical utility, particularly in global primary care settings where there is the greatest opportunity to identify people who need mental health treatment. Based on WHO's mission and constitution, the usefulness of the classification in helping WHO member countries, particularly low- and middle-income countries, to reduce the disease burden associated with mental disorders is among the highest priorities for the revision. This article describes the foundation provided by the recommendations of the Advisory Group for the current phase of work. PMID- 21633678 TI - WPA guidance on the protection and promotion of mental health in children of persons with severe mental disorders. AB - This guidance details the needs of children, and the qualities of parenting that meet those needs. Parental mental disorders can damage the foetus during pregnancy through the action of drugs, prescribed or abused. Pregnancy and the puerperium can exacerbate or initiate mental illness in susceptible women. After their birth, the children may suffer from the social disadvantage associated with severe mental illness. The parents (depending on the disorder, its severity and its persistence) may have intermittent or prolonged difficulties with parenting, which may sometimes result in childhood psychological disturbance or child maltreatment. This guidance considers ways of preventing, minimizing and remedying these effects. Our recommendations include: education of psychiatrists and related professions about the effect of parental mental illness on children; revision of psychiatric training to increase awareness of patients as caregivers, and to incorporate relevant assessment and intervention into their treatment and rehabilitation; the optimum use of pharmacological treatment during pregnancy; pre-birth planning when women with severe mental illness become pregnant; development of specialist services for pregnant and puerperal women, with assessment of their efficacy; community support for parenting by mothers and fathers with severe mental disorders; standards of good practice for the management of child maltreatment when parents suffer from mental illness; the importance of multi-disciplinary teamwork when helping these families, supporting their children and ensuring child protection; the development of child and adolescent mental health services worldwide. PMID- 21633679 TI - Personality and psychopathology. AB - PERSONALITY AND PSYCHOPATHOLOGY CAN RELATE TO ONE ANOTHER IN THREE DIFFERENT WAYS: personality and psychopathology can influence the presentation or appearance of one another (pathoplastic relationships); they can share a common, underlying etiology (spectrum relationships); and they can have a causal role in the development or etiology of one another. Each of these possible forms of inter relationship is considered in this paper. PMID- 21633680 TI - Further thoughts on the relation of personality and psychopathology. PMID- 21633681 TI - Personality diathesis explains the interrelationships between personality disorder and other mental conditions. PMID- 21633682 TI - Personality and psychopathology: the dangers of premature closure. PMID- 21633683 TI - Exploring personality - personality disorder relations and their implications for DSM-5. PMID- 21633684 TI - Interaction of personality disorders with other co-occurring psychiatric disorders. PMID- 21633685 TI - Personality may be psychopathology, and vice versa. PMID- 21633686 TI - Challenges in the study of personality and psychopathology. PMID- 21633687 TI - The influence of personality on the treatment outcome of psychopathology. PMID- 21633688 TI - Towards an approach to mental disorders based on individual differences. PMID- 21633689 TI - The WPA-WHO Global Survey of Psychiatrists' Attitudes Towards Mental Disorders Classification. AB - This article describes the results of the WPA-WHO Global Survey of 4,887 psychiatrists in 44 countries regarding their use of diagnostic classification systems in clinical practice, and the desirable characteristics of a classification of mental disorders. The WHO will use these results to improve the clinical utility of the ICD classification of mental disorders through the current ICD-10 revision process. Participants indicated that the most important purposes of a classification are to facilitate communication among clinicians and to inform treatment and management. They overwhelmingly preferred a simpler system with 100 or fewer categories, and over two-thirds preferred flexible guidance to a strict criteria-based approach. Opinions were divided about how to incorporate severity and functional status, while most respondents were receptive to a system that incorporates a dimensional component. Significant minorities of psychiatrists in Latin America and Asia reported problems with the cross-cultural applicability of existing classifications. Overall, ratings of ease of use and goodness of fit for specific ICD-10 categories were fairly high, but several categories were described as having poor utility in clinical practice. This represents an important focus for the ICD revision, as does ensuring that the ICD 11 classification of mental disorders is acceptable to psychiatrists throughout the world. PMID- 21633690 TI - Randomized controlled trial of supported employment in England: 2 year follow-up of the Supported Work and Needs (SWAN) study. AB - Studies from North America have concluded that supported employment using the Individual Placement and Support (IPS) model is effective in helping individuals with severe and persistent mental illness gain competitive employment. The aim of this study was to investigate the effectiveness and cost-effectiveness of IPS in England in patients followed up for 2 years. Patients with severe mental illness were randomised to IPS or local vocational services (treatment as usual). Service use and costs were measured. Two hundred-nineteen participants were randomised, and 86% re-assessed 2 years later. In the multivariate analysis, relatively low rates of competitive employment were found in both the intervention group and the treatment as usual group, although significantly more patients obtained competitive employment in the treatment arm (22% vs. 11%, p=0.041). There were no significant differences in costs. The employment rate among participants receiving IPS was lower than in previously published reports, and the number needed to treat to obtain the benefit of IPS was relatively high. This may reflect difficulties in the implementation of IPS where it is not structurally integrated within mental health teams, as well as economic disincentives which lead to lower levels of motivation for patients and mental health professionals. PMID- 21633692 TI - Bipolar units and programmes: are they really needed? PMID- 21633693 TI - Outcomes and future directions of the national community mental health care program in Viet Nam. PMID- 21633691 TI - Physical illness in patients with severe mental disorders. II. Barriers to care, monitoring and treatment guidelines, plus recommendations at the system and individual level. AB - Physical disorders are, compared to the general population, more prevalent in people with severe mental illness (SMI). Although this excess morbidity and mortality is largely due to modifiable lifestyle risk factors, the screening and assessment of physical health aspects remains poor, even in developed countries. Moreover, specific patient, provider, treatment and system factors act as barriers to the recognition and to the management of physical diseases in people with SMI. Psychiatrists can play a pivotal role in the improvement of the physical health of these patients by expanding their task from clinical psychiatric care to the monitoring and treatment of crucial physical parameters. At a system level, actions are not easy to realize, especially for developing countries. However, at an individual level, even simple and very basic monitoring and treatment actions, undertaken by the treating clinician, can already improve the problem of suboptimal medical care in this population. Adhering to monitoring and treatment guidelines will result in a substantial enhancement of physical health outcomes. Furthermore, psychiatrists can help educate and motivate people with SMI to address their suboptimal lifestyle, including smoking, unhealthy diet and lack of exercise. The adoption of the recommendations presented in this paper across health care systems throughout the world will contribute to a significant improvement in the medical and related psychiatric health outcomes of patients with SMI. PMID- 21633694 TI - Physical complications of severe enduring obsessive-compulsive disorder. PMID- 21633695 TI - WPA recommendations for relationships of psychiatrists, health care organizations working in the psychiatric field and psychiatric associations with the pharmaceutical industry. PMID- 21633696 TI - The 15th World Congress of Psychiatry and recent WPA activities. PMID- 21633697 TI - Activities of the WPA Early Career Psychiatrists Council: the Action Plan is in progress. PMID- 21633698 TI - Epidemiology, quality and reporting characteristics of systematic reviews of traditional Chinese medicine interventions published in Chinese journals. AB - BACKGROUND: Systematic reviews (SRs) of TCM have become increasingly popular in China and have been published in large numbers. This review provides the first examination of epidemiological characteristics of these SRs as well as compliance with the PRISMA and AMSTAR guidelines. OBJECTIVES: To examine epidemiological and reporting characteristics as well as methodological quality of SRs of TCM published in Chinese journals. METHODS: Four Chinese databases were searched (CBM, CSJD, CJFD and Wanfang Database) for SRs of TCM, from inception through Dec 2009. Data were extracted into Excel spreadsheets. The PRISMA and AMSTAR checklists were used to assess reporting characteristics and methodological quality, respectively. RESULTS: A total of 369 SRs were identified, most (97.6%) of which used the terms systematic review or meta-analysis in the title. None of the reviews had been updated. Half (49.8%) were written by clinicians and nearly half (47.7%) were reported in specialty journals. The impact factors of 45.8% of the journals published in were zero. The most commonly treated conditions were diseases of the circulatory and digestive disease. Funding sources were not reported for any reviews. Most (68.8%) reported information about quality assessment, while less than half (43.6%) reported assessing for publication bias. Statistical mistakes appeared in one-third (29.3%) of reviews and most (91.9%) did not report on conflict of interest. CONCLUSIONS: While many SRs of TCM interventions have been published in Chinese journals, the quality of these reviews is troubling. As a potential key source of information for clinicians and researchers, not only were many of these reviews incomplete, some contained mistakes or were misleading. Focusing on improving the quality of SRs of TCM, rather than continuing to publish them in great quantity, is urgently needed in order to increase the value of these studies. PMID- 21633699 TI - A role for hemopexin in oligodendrocyte differentiation and myelin formation. AB - Myelin formation and maintenance are crucial for the proper function of the CNS and are orchestrated by a plethora of factors including growth factors, extracellular matrix components, metalloproteases and protease inhibitors. Hemopexin (Hx) is a plasma protein with high heme binding affinity, which is also locally produced in the CNS by ependymal cells, neurons and glial cells. We have recently reported that oligodendrocytes (OLs) are the type of cells in the brain that are most susceptible to lack of Hx, as the number of iron-overloaded OLs increases in Hx-null brain, leading to oxidative tissue damage. In the current study, we found that the expression of the Myelin Basic Protein along with the density of myelinated fibers in the basal ganglia and in the motor and somatosensory cortex of Hx-null mice were strongly reduced starting at 2 months and progressively decreased with age. Myelin abnormalities were confirmed by electron microscopy and, at the functional level, resulted in the inability of Hx null mice to perform efficiently on the Rotarod. It is likely that the poor myelination in the brain of Hx-null mice was a consequence of defective maturation of OLs as we demonstrated that the number of mature OLs was significantly reduced in mutant mice whereas that of precursor cells was normal. Finally, in vitro experiments showed that Hx promotes OL differentiation. Thus, Hx may be considered a novel OL differentiation factor and the modulation of its expression in CNS may be an important factor in the pathogenesis of human neurodegenerative disorders. PMID- 21633700 TI - T cells contribute to tumor progression by favoring pro-tumoral properties of intra-tumoral myeloid cells in a mouse model for spontaneous melanoma. AB - Tumors affect myelopoeisis and induce the expansion of myeloid cells with immunosuppressive activity. In the MT/ret model of spontaneous metastatic melanoma, myeloid cells are the most abundant tumor infiltrating hematopoietic population and their proportion is highest in the most aggressive cutaneous metastasis. Our data suggest that the tumor microenvironment favors polarization of myeloid cells into type 2 cells characterized by F4/80 expression, a weak capacity to secrete IL-12 and a high production of arginase. Myeloid cells from tumor and spleen of MT/ret mice inhibit T cell proliferation and IFNgamma secretion. Interestingly, T cells play a role in type 2 polarization of myeloid cells. Indeed, intra-tumoral myeloid cells from MT/ret mice lacking T cells are not only less suppressive towards T cells than corresponding cells from wild-type MT/ret mice, but they also inhibit more efficiently melanoma cell proliferation. Thus, our data support the existence of a vicious circle, in which T cells may favor cancer development by establishing an environment that is likely to skew myeloid cell immunity toward a tumor promoting response that, in turn, suppresses immune effector cell functions. PMID- 21633701 TI - Gbetagamma and the C terminus of SNAP-25 are necessary for long-term depression of transmitter release. AB - BACKGROUND: Short-term presynaptic inhibition mediated by G protein-coupled receptors involves a direct interaction between G proteins and the vesicle release machinery. Recent studies implicate the C terminus of the vesicle associated protein SNAP-25 as a molecular binding target of Gbetagamma that transiently reduces vesicular release. However, it is not known whether SNAP-25 is a target for molecular modifications expressing long-term changes in transmitter release probability. METHODOLOGY/PRINCIPAL FINDINGS: This study utilized two-photon laser scanning microscopy for real-time imaging of action potential-evoked [Ca(2+)] increases, in single Schaffer collateral presynaptic release sites in in vitro hippocampal slices, plus simultaneous recording of Schaffer collateral-evoked synaptic potentials. We used electroporation to infuse small peptides through CA3 cell bodies into presynaptic Schaffer collateral terminals to selectively study the presynaptic effect of scavenging the G-protein Gbetagamma. We demonstrate here that the C terminus of SNAP-25 is necessary for expression of LTD, but not long-term potentiation (LTP), of synaptic strength. Using type A botulinum toxin (BoNT/A) to enzymatically cleave the 9 amino acid C terminus of SNAP-25 eliminated the ability of low frequency synaptic stimulation to induce LTD, but not LTP, even if release probability was restored to pre BoNT/A levels by elevating extracellular [Ca(2+)]. Presynaptic electroporation infusion of the 14-amino acid C-terminus of SNAP-25 (Ct-SNAP-25), to scavenge Gbetagamma, reduced both the transient presynaptic inhibition produced by the group II metabotropic glutamate receptor stimulation, and LTD. Furthermore, presynaptic infusion of mSIRK, a second, structurally distinct Gbetagamma scavenging peptide, also blocked the induction of LTD. While Gbetagamma binds directly to and inhibit voltage-dependent Ca(2+) channels, imaging of presynaptic [Ca(2+)] with Mg-Green revealed that low-frequency stimulation only transiently reduced presynaptic Ca(2+) influx, an effect not altered by infusion of Ct-SNAP 25. CONCLUSIONS/SIGNIFICANCE: The C-terminus of SNAP-25, which links synaptotagmin I to the SNARE complex, is a binding target for Gbetagamma necessary for both transient transmitter-mediated presynaptic inhibition, and the induction of presynaptic LTD. PMID- 21633702 TI - Rapid evolution of coral proteins responsible for interaction with the environment. AB - BACKGROUND: Corals worldwide are in decline due to climate change effects (e.g., rising seawater temperatures), pollution, and exploitation. The ability of corals to cope with these stressors in the long run depends on the evolvability of the underlying genetic networks and proteins, which remain largely unknown. A genome wide scan for positively selected genes between related coral species can help to narrow down the search space considerably. METHODOLOGY/PRINCIPAL FINDINGS: We screened a set of 2,604 putative orthologs from EST-based sequence datasets of the coral species Acropora millepora and Acropora palmata to determine the fraction and identity of proteins that may experience adaptive evolution. 7% of the orthologs show elevated rates of evolution. Taxonomically-restricted (i.e. lineage-specific) genes show a positive selection signature more frequently than genes that are found across many animal phyla. The class of proteins that displayed elevated evolutionary rates was significantly enriched for proteins involved in immunity and defense, reproduction, and sensory perception. We also found elevated rates of evolution in several other functional groups such as management of membrane vesicles, transmembrane transport of ions and organic molecules, cell adhesion, and oxidative stress response. Proteins in these processes might be related to the endosymbiotic relationship corals maintain with dinoflagellates in the genus Symbiodinium. CONCLUSION/RELEVANCE: This study provides a birds-eye view of the processes potentially underlying coral adaptation, which will serve as a foundation for future work to elucidate the rates, patterns, and mechanisms of corals' evolutionary response to global climate change. PMID- 21633703 TI - Angelman syndrome protein UBE3A interacts with primary microcephaly protein ASPM, localizes to centrosomes and regulates chromosome segregation. AB - Many proteins associated with the phenotype microcephaly have been localized to the centrosome or linked to it functionally. All the seven autosomal recessive primary microcephaly (MCPH) proteins localize at the centrosome. Microcephalic osteodysplastic primordial dwarfism type II protein PCNT and Seckel syndrome (also characterized by severe microcephaly) protein ATR are also centrosomal proteins. All of the above findings show the importance of centrosomal proteins as the key players in neurogenesis and brain development. However, the exact mechanism as to how the loss-of-function of these proteins leads to microcephaly remains to be elucidated. To gain insight into the function of the most commonly mutated MCPH gene ASPM, we used the yeast two-hybrid technique to screen a human fetal brain cDNA library with an ASPM bait. The analysis identified Angelman syndrome gene product UBE3A as an ASPM interactor. Like ASPM, UBE3A also localizes to the centrosome. The identification of UBE3A as an ASPM interactor is not surprising as more than 80% of Angelman syndrome patients have microcephaly. However, unlike in MCPH, microcephaly is postnatal in Angelman syndrome patients. Our results show that UBE3A is a cell cycle regulated protein and its level peaks in mitosis. The shRNA knockdown of UBE3A in HEK293 cells led to many mitotic abnormalities including chromosome missegregation, abnormal cytokinesis and apoptosis. Thus our study links Angelman syndrome protein UBE3A to ASPM, centrosome and mitosis for the first time. We suggest that a defective chromosome segregation mechanism is responsible for the development of microcephaly in Angelman syndrome. PMID- 21633704 TI - Validation of the tetracycline regulatable gene expression system for the study of the pathogenesis of infectious disease. AB - Understanding the pathogenesis of infectious disease requires the examination and successful integration of parameters related to both microbial virulence and host responses. As a practical and powerful method to control microbial gene expression, including in vivo, the tetracycline-regulatable system has recently gained the favor of many investigative groups. However, some immunomodulatory effects of the tetracyclines, including doxycycline, could potentially limit its use to evaluate host responses during infection. Here we have used a well established murine model of disseminated candidiasis, which is highly dependent on both the virulence displayed by the fungal cells and on the host immune status, to validate the use of this system. We demonstrate that the pathogenesis of the wild type C. albicans CAF2-1 strain, which does not contain any tet regulatable element, is not affected by the presence of doxycycline. Moreover levels of key cytokines, chemokines and many other biomarkers, as determined by multi-analyte profiling, remain essentially unaltered by the presence of the antibiotic during infection. Our results indicate that the levels of doxycycline needed to control the tetracycline regulatable promoter gene expression system have no detectable effect on global host responses during candidiasis. Because tet-regulatable systems are now being increasingly used in a variety of pathogenic microorganisms, these observations have wide implications in the field of infectious diseases. PMID- 21633705 TI - Tyrosine sulfation of native mouse Psgl-1 is required for optimal leukocyte rolling on P-selectin in vivo. AB - BACKGROUND: We recently demonstrated that tyrosine sulfation is an important contributor to monocyte recruitment and retention in a mouse model of atherosclerosis. P-selectin glycoprotein ligand-1 (Psgl-1) is tyrosine-sulfated in mouse monocyte/macrophages and its interaction with P-selectin is important in monocyte recruitment in atherosclerosis. However, whether tyrosine sulfation is required for the P-selectin binding function of mouse Psgl-1 is unknown. Here we test the function of native Psgl-1 expressed in leukocytes lacking endogenous tyrosylprotein sulfotransferase (TPST) activity. METHODOLOGY/PRINCIPAL FINDINGS: Psgl-1 function was assessed by examining P-selectin dependent leukocyte rolling in post-capillary venules of C57BL6 mice transplanted with hematopoietic progenitors from wild type (WT -> B6) or Tpst1;Tpst2 double knockout mice (Tpst DKO -> B6) which lack TPST activity. We observed that rolling flux fractions were lower and leukocyte rolling velocities were higher in Tpst DKO -> B6 venules compared to WT -> B6 venules. Similar results were observed on immobilized P selectin in vitro. Finally, Tpst DKO leukocytes bound less P-selectin than wild type leukocytes despite equivalent surface expression of Psgl-1. CONCLUSIONS/SIGNIFICANCE: These findings provide direct and convincing evidence that tyrosine sulfation is required for optimal function of mouse Psgl-1 in vivo and suggests that tyrosine sulfation of Psgl-1 contributes to the development of atherosclerosis. PMID- 21633706 TI - Repair at single targeted DNA double-strand breaks in pluripotent and differentiated human cells. AB - Differences in ex vivo cell culture conditions can drastically affect stem cell physiology. We sought to establish an assay for measuring the effects of chemical, environmental, and genetic manipulations on the precision of repair at a single DNA double-strand break (DSB) in pluripotent and somatic human cells. DSBs in mammalian cells are primarily repaired by either homologous recombination (HR) or nonhomologous end-joining (NHEJ). For the most part, previous studies of DSB repair in human cells have utilized nonspecific clastogens like ionizing radiation, which are highly nonphysiologic, or assayed repair at randomly integrated reporters. Measuring repair after random integration is potentially confounded by locus-specific effects on the efficiency and precision of repair. We show that the frequency of HR at a single DSB differs up to 20-fold between otherwise isogenic human embryonic stem cells (hESCs) based on the site of the DSB within the genome. To overcome locus-specific effects on DSB repair, we used zinc finger nucleases to efficiently target a DSB repair reporter to a safe harbor locus in hESCs and a panel of somatic human cell lines. We demonstrate that repair at a targeted DSB is highly precise in hESCs, compared to either the somatic human cells or murine embryonic stem cells. Differentiation of hESCs harboring the targeted reporter into astrocytes reduces both the efficiency and precision of repair. Thus, the phenotype of repair at a single DSB can differ based on either the site of damage within the genome or the stage of cellular differentiation. Our approach to single DSB analysis has broad utility for defining the effects of genetic and environmental modifications on repair precision in pluripotent cells and their differentiated progeny. PMID- 21633707 TI - L,L-diaminopimelate aminotransferase from Chlamydomonas reinhardtii: a target for algaecide development. AB - In some bacterial species and photosynthetic cohorts, including algae, the enzyme L,L-diaminopimelate aminotransferase (DapL) (E.C. 2.6.1.83) is involved in the anabolism of the essential amino acid L-lysine. DapL catalyzes the conversion of tetrahydrodipicolinate (THDPA) to L,L-diaminopimelate (L,L-DAP), in one step bypassing the DapD, DapC and DapE enzymatic reactions present in the acyl DAP pathways. Here we present an in vivo and in vitro characterization of the DapL ortholog from the alga Chlamydomonas reinhardtii (Cr-DapL). The in vivo analysis illustrated that the enzyme is able to functionally complement the E. coli dap auxotrophs and was essential for plant development in Arabidopsis. In vitro, the enzyme was able to inter-convert THDPA and L,L-DAP, showing strong substrate specificity. Cr-DapL was dimeric in both solution and when crystallized. The structure of Cr-DapL was solved in its apo form, showing an overall architecture of a alpha/beta protein with each monomer in the dimer adopting a pyridoxal phosphate-dependent transferase-like fold in a V-shaped conformation. The active site comprises residues from both monomers in the dimer and shows some rearrangement when compared to the apo-DapL structure from Arabidopsis. Since animals do not possess the enzymatic machinery necessary for the de novo synthesis of the amino acid L-lysine, enzymes involved in this pathway are attractive targets for the development of antibiotics, herbicides and algaecides. PMID- 21633708 TI - Low vitamin D in narcolepsy with cataplexy. AB - BACKGROUND: Narcolepsy with cataplexy (NC) is currently thought to be an autoimmune-mediated disorder in which environmental risk factors make a significant contribution to its development. It was proposed that vitamin D deficiency plays a role in autoimmune diseases. Here we investigated whether NC can be associated with 25-hydroxyvitamin D (25(OH)D) level deficiency in patients with NC compared with gender- and age-matched normal controls. METHODOLOGY: Serum level of 25 (OH)D was determined in 51 European patients with typical NC compared to 55 age-, gender-, and ethnicity-matched healthy controls. Demographic and clinical data (age at onset, duration and severity of disease at baseline, and treatment intake at time of study) and season of blood sampling were collected to control for confounding variables. PRINCIPAL FINDINGS: Serum 25(OH)D concentration was lower in NC compared to controls (median, 59.45 nmol/l [extreme values 24.05-124.03] vs. 74.73 nmol/l [26.88-167.48] p = 0.0039). Patients with NC had significantly greater vitamin D deficiency (<75 nmol/l) than controls (72.5% vs 50.9%, p = 0.0238). Division into quartiles of the whole sample revealed that the risk of being affected with NC increased with lower 25(OH)D level, with a 5.34 OR [1.65-17.27] for the lowest quartile (p = 0.0051). Further adjustment for BMI did not modify the strength of the association (OR: 3.63, 95% CI = 1.06-12.46, p = 0.0191). No between BMI and 25(OH)D interaction, and no correlation between 25(OH)D level and disease duration or severity or treatment intake were found in NC. CONCLUSION: We found a higher frequency of vitamin D deficiency in NC. Further studies are needed to assess the contribution of hypovitaminosis D to the risk of developing narcolepsy, and to focus on the utility of assessing vitamin D status to correct potential deficiency. PMID- 21633710 TI - Run-on mutation in the PAX6 gene and chorioretinal degeneration in autosomal dominant aniridia. AB - PURPOSE: To identify the causative paired box 6 (PAX6) mutation in a family with autosomal dominant aniridia. METHODS: A family with autosomal dominant aniridia with three affected individuals in two generations was investigated for the causative PAX6 mutation by single strand conformation polymorphism (SSCP) followed by sequencing of genomic DNA from peripheral blood. RESULTS: A novel PAX6 mutation in the donor splice site of intron 12 was identified in all three affected individuals from the family. The automated splice site analysis web interface indicated a disturbance of splicing and it was predicted that this mutation could lead to an elimination of the normal stop codon and an abnormal 3' elongation of the mRNA. CONCLUSIONS: We report a novel PAX6 mutation in autosomal dominant aniridia that presumably affects splicing. The presence of chorioretinal degeneration in one of the affected individual raises the possibility that run-on mutations are associated with chorioretinal involvement in aniridia. PMID- 21633709 TI - Sequencing and comparative genome analysis of two pathogenic Streptococcus gallolyticus subspecies: genome plasticity, adaptation and virulence. AB - Streptococcus gallolyticus infections in humans are often associated with bacteremia, infective endocarditis and colon cancers. The disease manifestations are different depending on the subspecies of S. gallolyticus causing the infection. Here, we present the complete genomes of S. gallolyticus ATCC 43143 (biotype I) and S. pasteurianus ATCC 43144 (biotype II.2). The genomic differences between the two biotypes were characterized with comparative genomic analyses. The chromosome of ATCC 43143 and ATCC 43144 are 2,36 and 2,10 Mb in length and encode 2246 and 1869 CDS respectively. The organization and genomic contents of both genomes were most similar to the recently published S. gallolyticus UCN34, where 2073 (92%) and 1607 (86%) of the ATCC 43143 and ATCC 43144 CDS were conserved in UCN34 respectively. There are around 600 CDS conserved in all Streptococcus genomes, indicating the Streptococcus genus has a small core-genome (constitute around 30% of total CDS) and substantial evolutionary plasticity. We identified eight and five regions of genome plasticity in ATCC 43143 and ATCC 43144 respectively. Within these regions, several proteins were recognized to contribute to the fitness and virulence of each of the two subspecies. We have also predicted putative cell-surface associated proteins that could play a role in adherence to host tissues, leading to persistent infections causing sub-acute and chronic diseases in humans. This study showed evidence that the S. gallolyticus still possesses genes making it suitable in a rumen environment, whereas the ability for S. pasteurianus to live in rumen is reduced. The genome heterogeneity and genetic diversity among the two biotypes, especially membrane and lipoproteins, most likely contribute to the differences in the pathogenesis of the two S. gallolyticus biotypes and the type of disease an infected patient eventually develops. PMID- 21633711 TI - Mast cells modulate the inflammatory process in endotoxin-induced uveitis. AB - PURPOSE: To investigate the role of mast cells and annexin-A1 (Anxa1) in endotoxin-induced uveitis (EIU). METHODS: EIU was induced by injection of lipopolysaccharide (LPS) into the paws of rats, which were then sacrificed after 24 and 48 h. To assess EIU in the absence of mast cells, groups of animals were pretreated with compound 48/80 (c48/80) and sacrificed after 24 h after no treatment or EIU induction. The eyes were used for histological studies and the aqueous humor (AqH) pool was used for the analysis of transmigrated cells and Anxa1 levels. In inflammatory cells, Anxa1 expression was monitored by immunohistochemistry. RESULTS: After 24 h, rats with EIU exhibited degranulated mast cells, associated with elevated numbers of infiltrating leukocytes and the high expression of Anxa1 in the AqH and the neutrophils. After 48 h of EIU, the mast cells were intact, indicating granule re-synthesis, and there was a reduction of neutrophil transmigration and an increase in the number of mononuclear phagocytic cells in ocular tissues. Anxa1 expression was decreased in neutrophils but increased in mononuclear phagocytic cells. In the animals pretreated with c48/80 and subjected to EIU, mast cells responded to this secretagogue by degranulating and few transmigrated neutrophils were observed. CONCLUSTIONS: We report that mast cells are a potential source of pharmacological mediators that are strongly linked to the pathophysiology of EIU, and the endogenous protein Anxa1 is a mediator in the homeostasis of the inflammatory process with anti-migratory effects on leukocytes, which supports further studies of this protein as an innovative therapy for uveitis. PMID- 21633712 TI - A novel 1-bp deletion in PITX3 causing congenital posterior polar cataract. AB - PURPOSE: Cataracts are the most common cause of blindness worldwide. Inherited cataract is a clinically and genetically heterogeneous disease. Here we report a novel mutation in the paired-like homeodomain 3 (PITX3) gene segregating in a four generation English family with an isolated autosomal dominant posterior polar cataract. METHODS: A genome-wide linkage was performed by means of single nucleotide polymorphism (SNP) and microsatellite markers. Linkage analyses were performed with the GeneHunter and MLINK programs. Direct sequencing of PCR products was performed to detect mutation in the gene, using the BigDye version 3.1 and analyzed using Sequence analysis version 5.2. RESULTS: Genome-wide linkage analysis with SNP markers, identified a disease-haplotype interval on chromosome 10q. Two point positive logarithm of odds (LOD) scores was obtained with markers D10S205 (Z=3.10 at theta=0.00), flanked by markers D10S1709 and D10S543, which harbors the homeobox gene PITX3. Sequence analysis of PITX3 revealed a 1-bp deletion that cosegregated with all the affected members of this family which resulted in a frameshift in codon 181 and likely to produce an aberrant protein consisting of 127 additional residues. CONCLUSIONS: The 542delC is a novel mutation in PITX3 causing an isolated posterior polar cataract. PMID- 21633713 TI - Inhalative preconditioning with hydrogen sulfide attenuated apoptosis after retinal ischemia/reperfusion injury. AB - PURPOSE: Retinal ischemia/reperfusion (I/R) injury plays an important role in the pathophysiology of various ocular diseases. Retinal ganglion cells (RGCs) are particularly vulnerable to ischemia. Hydrogen sulfide (H(2)S) was recently shown to be neuroprotective in the brain and retina due to its antiapoptotic effects. Rapid preconditioning of retinal neurons by inhaled H(2)S before I/R injury may reduce apoptosis in the rat retina. METHODS: I/R injury was created on the left eye of rats (n=8) with or without inhaled H(2)S preconditioning (80 ppm) for one hour before ischemia. Densities of fluorogold prelabeled RGCs were analyzed 7 days after injury in retinal whole mounts. Retinal tissue was harvested to analyze protein expression of heat shock protein (HSP)-90 and the mitogen activated protein kinases (MAPKs) c-jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK)1/2 and p38 to elucidate a possible pathway of neuroprotection. DNA binding activity of the transcription factors nuclear factor kappa-light-chain-enhancer of activated B-cells (NF-kappaB), cyclic adenosine monophosphate response element binding protein (CREB), and heat shock element (HSE), as well as caspase-3 cleavage and activity, were determined. Retinal sections were further assessed using anti-glial fibrillary acidic protein staining. RESULTS: RGC death after I/R injury decreased by 41.5% after H(2)S preconditioning compared to room air (p<0.001). H(2)S inhalation before ischemia reduced caspase-3 cleavage (p<0.001) and attenuated caspase-3 activity (p<0.001). Furthermore, HSP-90 expression was significantly elevated in the retina after H(2)S preconditioning. NF-kappaB but not CREB or HSE showed specific, H2S dependent regulation, as well as the MAPKs ERK1/2 and JNK but not p38. CONCLUSIONS: H(2)S preconditioning mediates antiapoptotic effects in retinal I/R injury, thus exhibiting neuroprotection. Based on these observations, H(2)S could represent a novel and promising therapeutic agent to counteract neuronal injuries in the eye. Further studies are needed to prove H(2)S's neuroprotective propensity using a postconditioning approach. PMID- 21633714 TI - Altered expression of beta-galactosidase-1-like protein 3 (Glb1l3) in the retinal pigment epithelium (RPE)-specific 65-kDa protein knock-out mouse model of Leber's congenital amaurosis. AB - PURPOSE: In this study, we investigated the expression of the gene encoding beta galactosidase (Glb)-1-like protein 3 (Glb1l3), a member of the glycosyl hydrolase 35 family, during retinal degeneration in the retinal pigment epithelium (RPE) specific 65-kDa protein knockout (Rpe65(-/-)) mouse model of Leber congenital amaurosis (LCA). Additionally, we assessed the expression of the other members of this protein family, including beta-galactosidase-1 (Glb1), beta-galactosidase-1 like (Glb1l), and beta-galactosidase-1-like protein 2 (Glb1l2). METHODS: The structural features of Glb1l3 were assessed using bioinformatic tools. mRNA expression of Glb-related genes was investigated by oligonucleotide microarray, real-time PCR, and reverse transcription (RT) -PCR. The localized expression of Glb1l3 was assessed by combined in situ hybridization and immunohistochemistry. RESULTS: Glb1l3 was the only Glb-related member strongly downregulated in Rpe65( /-) retinas before the onset and during progression of the disease. Glb1l3 mRNA was only expressed in the retinal layers and the RPE/choroid. The other Glb related genes were ubiquitously expressed in different ocular tissues, including the cornea and lens. In the healthy retina, expression of Glb1l3 was strongly induced during postnatal retinal development; age-related increased expression persisted during adulthood and aging. CONCLUSIONS: These data highlight early onset downregulation of Glb1l3 in Rpe65-related disease. They further indicate that impaired expression of Glb1l3 is mostly due to the absence of the chromophore 11-cis retinal, suggesting that Rpe65 deficiency may have many metabolic consequences in the underlying neuroretina. PMID- 21633715 TI - Age-related alterations in retinal neurovascular and inflammatory transcripts. AB - PURPOSE: Vision loss is one of the most common complications of aging, even in individuals with no diagnosed ocular disease. Increasing age induces structural alterations and functional impairments in retinal neurons and microvasculature linked to the activation of proinflammatory signaling pathways. Commonalities between the effects of aging and those observed with diabetes, including visual impairment, vascular dysfunction, and increased inflammatory response, have led to the hypothesis that diabetes-associated pathologies reflect an "advanced aging" phenotype. The goal of this study was to investigate the effects of aging on retinal mRNA expression of neurovascular and inflammatory transcripts previously demonstrated to be regulated with diabetes. METHODS: The relative expression of 36 genes of interest previously identified as consistently regulated with diabetes was assessed in retinas of Young (3 month), Adult (12 month), and Aged (26 month) Fischer 344 x Brown Norway (F1) hybrid rats using quantitative PCR. Serum samples obtained at sacrifice were assayed to determine serum glucose levels. RESULTS: Eleven inflammation- and microvascular-related genes previously demonstrated to be upregulated in young diabetic rats (complement component 1 s subcomponent [C1s], chitinase 3-like 1 [Chi3L1], endothelin 2 [Edn2], guanylate nucleotide binding protein 2 [Gbp2], glial fibrillary acidic protein [Gfap], intracellular adhesion molecule 1 [Icam1], janus kinase 3 [Jak3], lipopolysaccharide-induced TNF factor [Litaf], complement 1-inhibitor [Serping1], signal transducer and activator of transcription 3 [Stat3], tumor necrosis factor receptor subfamily member 12a [Tnfrsf12a]) demonstrated progressively increasing retinal expression in aged normoglycemic rats. Additionally, two neuronal function-related genes (glutamate receptor ionotropic NMDA 2A [Grin2a] and polycomb group ring finger 1 [Pcgf1]) and one inflammation-related gene (pigment epithelium-derived growth factor [Pedf]) displayed patterns of expression dissimilar to that previously demonstrated with diabetes. CONCLUSIONS: The commonalities in retinal age-related and diabetes induced molecular alterations provide support for the hypothesis that diabetes and aging engage some common para-inflammatory processes. However, these results also demonstrate that while the retinal genomic response to diabetes and aging share commonalities, they are not superimposable phenotypes. The observed changes in retinal gene expression provide further evidence of retinal alterations in neurovascular and inflammatory processes across the adult rat lifespan; this is indicative of para-inflammation that may contribute to the functional impairments that occur with advanced age. The data also suggest the potential for an additive effect of aging and diabetes in the development of diabetic complications. PMID- 21633716 TI - Lipid peroxidation and total antioxidant capacity in vitreous, aqueous humor, and blood samples from patients with diabetic retinopathy. AB - PURPOSE: To evaluate levels of malondialdehyde and the total antioxidant capacity (TAC) in the blood, aqueous humor, and vitreous bodies of diabetic and nondiabetic patients. We also measured the blood energy charge potential (ECP). METHODS: We examined 19 patients with type 2 diabetes mellitus and diabetic retinopathy. Ten were scheduled for cataract surgery and pars plana vitrectomy because of proliferative diabetic retinopathy (PDR). The other nine, with mild nonproliferative PDR (NPDR), and fourteen nondiabetic, age-matched subjects enrolled as a control group were scheduled for cataract surgery and vitrectomy because of epiretinal membranes. Blood, aqueous humor and vitreous body samples were collected at the time of surgery. Malondialdehyde concentrations and blood ECP were measured with high-performance liquid chromatography. The TAC of the samples was estimated with the oxygen radical absorbance capacity method. RESULTS: The level of blood and vitreous malondialdehyde in the PDR group was significantly higher compared to controls and to NPDR patients. PDR patients also had lower levels of TAC at the vitreous body and aqueous humor level, but not at the blood level, compared to controls and with NPDR patients. In all diabetic patients, the blood ECP values were significantly lower, compared to control subjects. CONCLUSIONS: Our data support the hypothesis that oxidative stress and the decrease of antioxidant defenses are associated with the progression of diabetic retinopathy to its proliferative form. Antioxidant supply may have the effect of correcting oxidative stress and inhibiting disease progression. PMID- 21633717 TI - Renin-angiotensin-aldosterone system genes and nonarteritic anterior ischemic optic neuropathy. AB - PURPOSE: Recent literature suggests a genetic component for non-arteritic anterior ischemic optic neuropathy (NAION). We examined the association of the insertion/deletion (I/D) polymorphism of the angiotensin-converting enzyme gene, of the M235T polymorphism of the angiotensinogen gene, and of the A1166C polymorphism of the angiotensin II type 1 receptor gene with NAION. METHODS: Forty-seven patients with NAION and 76 controls, age- and gender-matched, were recruited and genotyped for renin-angiotensin-aldosterone system (RAAS) genes. Genotypes were determined by polymerase chain reaction and restriction enzyme analysis. NAION and control groups were compared in regard to the prevalence of renin-angiotensin-aldosterone system polymorphisms, and further stratified by age and gender. RESULTS: NAION occurrence was not associated with the M235T polymorphism of the angiotensinogen gene and the A1166C polymorphism of the angiotensin II, type 1 receptor gene. Regarding the angiotensin-converting enzyme insertion/deletion polymorphism, our findings suggest that the II genotype could be a risk factor for NAION in younger male patients when compared to all cases and controls (p=0.033, odds ratio=5.71, confidence interval=1.152C28.35 and p=0.03, odds ratio=5.33, confidence interval=1.17C24.31 respectively). Furthermore I allele was present in all male patients younger than 55 years, making this allele a likely predisposing factor for NAION in young males. CONCLUSIONS: Since NAION may occur when compromised watershed microcirculation is combined with insufficient autoregulation of systematic circulation, polymorphisms of genes involved in systematic circulation, such as the RAAS genes, may be associated with NAION occurrence. Large-scale, multicentered, controlled prospective studies are needed to further explore the effects of RAAS polymorphisms or other genetic factors on NAION susceptibility. PMID- 21633718 TI - Pharmacoperone identification for therapeutic rescue of misfolded mutant proteins. AB - G protein-coupled receptors (GPCRs), which includes the gonadotropin releasing hormone (GnRH) receptor (GnRHR), comprises the largest family of validated drug targets-more than half of all approved drugs derive their benefits by selective targeting of GPCRs. Most drugs in this class are either agonists or antagonists of GPCRs and high throughput screens (HTSs) have typically been designed and performed with a view toward identification of such compounds as lead drug candidates. This manuscript presents the case that valuable drugs which effect the trafficking of GPCRs may have been overlooked because pharmacoperones have been selected from existing screens that identify agonists and antagonists. A "gain of activity assay" is proposed; this assay relies on the expression of a mutant of the GnRHR that is known to be rescuable by pharmacoperone drugs, and which is restored to activity in their presence. Accordingly, "hits" are identified by the appearance of activity. The gene for the mutant is under control of tetracycline and may be prevented from being expressed. This is a valuable feature since it allows false positives to be identified. Such drugs will show apparent activity whether or not the mutant is expressed. This assay will enable identification of these drugs from chemical libraries and does not rely on their activity as agonists or antagonists. PMID- 21633719 TI - Evaluation of fluticasone propionate and fluticasone propionate/salmeterol combination on exercise in pediatric and adolescent patients with asthma. AB - OBJECTIVE: This study was designed to demonstrate that four weeks of fluticasone propionate (FP) 100 micrograms (mcg) combined with salmeterol 50 mcg twice daily (BID) via DISKUS((r)) resulted in protection against bronchospasm induced by activity, as measured by standardized exercise challenge testing in pediatric and adolescent subjects who required regular use of inhaled corticosteroids for the treatment of persistent asthma. METHODS: Prior to study entry, all patients reported regular use of inhaled corticosteroids (ICS). During screening all patients demonstrated >=20% fall in FEV(1) following exercise. RESULTS: A total of 231 subjects aged 4 to 17 were randomized to the two study treatments: 113 to the FP/salmeterol combination group (FSC) and 118 to receive FP 100 mcg BID. Of the subjects randomized, 106 (94%) subjects in the FSC 100/50 group and 108 (92%) subjects in the FP 100 group completed the study. At the end of treatment (Week 4), both FSC and FP protected against a fall in FEV(1) following exercise in patients who at baseline experienced >=20% fall in FEV(1) following exercise. A mean decrease in FEV(1) of 9.9% was observed in the FSC 100/50 group as compared with a mean decrease of 11.1% in the FP 100 group; there was no statistical difference between treatments. CONCLUSION: Both FSC 100/50 and FP 100 provided protection against an exercised-induced fall in FEV(1); but statistically significant differences we not noted. Both treatments were well-tolerated over four weeks and FSC 100/50 had an adverse event profile comparable to that observed with FP 100. PMID- 21633720 TI - Effect of Guanabenz on Rat AMD Models and Rabbit Choroidal Blood Flow. AB - AIM: The effects of Guanabenz, an agonist of alpha2-adrenergic receptors routinely used in human medicine as an antihypertensive drug, were studied on NaIO(3)-induced retinal pigment epithelium (RPE) degeneration, laser-induced choroidal neovascularization (CNV) and choroidal blood flow, in animal models. METHODS: The 35mg/kg NaIO(3)-induced RPE degeneration rat eyes were instilled with 1% Guanabenz eye drops 3 times a day for 7 days before NaIO(3) injection, and then 2 to 4 weeks thereafter. RPE function was measured with c-wave of electroretinogram (ERG). Male Brown Norway rats were anesthetized to receive Nd:YAG laser to break the Bruch's membrane. One percent Guanabenz eye drops were given likewise. The development of CNV was determined by fluorescein angiography performed on week 2 and week 4 using sodium fluorescein (FA) or fluorescein isothiocyanatedextran (FD70-FA). Colored microsphere technique was used for in vivo experiments to determine the choroidal blood flow in ocular hypertensive (40 mmHg) rabbit eyes. RESULTS: The RPE function was protected significantly by Guanabenz according to the c-wave of ERG. Four weeks after NaIO(3) injection, the amplitude of ERG c-wave was 0.422+/-0.092 millivolts in the control group, 0.103+/-0.04 millivolts in the NaIO(3) group, and 0.254+/-0.061 millivolts in the Guanabenz+NaIO(3) group. There was a significant protection of the ERG c-wave by Guanabenz as compared to NaIO(3) group (P<0.01). The angiograms of FD70-FA showed decreased lesion size in the Guanabenz group. Four weeks after laser treatment, the size of the CNV lesion was 2.99+/-0.18 mm(2) in the control group, and 1.24+/ 0.16 mm(2) in the Guanabenz group (P<0.01). The choroidal blood flow was significantly increased at 30 and 60 minutes after Guanabenz instillation as compared to corresponding controls. CONCLUSIONS: Guanabenz significantly protected RPE from NaIO(3)-induced degeneration, inhibited the development of CNV in laser-induced rat AMD model and increased choroidal blood flow markedly in vivo. PMID- 21633721 TI - Coordination between nitric oxide and superoxide anion radical during progressive exercise in elite soccer players. AB - BACKGROUND: Exercise increases production of reactive oxygen and nitrogen species (RONS) via several mechanisms. Inter alia, increased blood flow during exercise exposes endothelial cells to shear stress, resulting in increased nitric oxide (NO) production. Increased oxygen consumption or hypoxia during exercise induces increased production of superoxide anion radical (O(2) (-)). OBJECTIVE: This study investigates the effects of maximal progressive treadmill exercise test on time-course of peripheral blood NO and O(2) (-) production, as well as the effect of long-term training on NO bioavailability. METHODS: Blood samples of 19 elite soccer players were gathered immediately before the test, during last 10 sec of every test stage, and during active recovery phases. RESULTS: Significant increase (p<0.05) in NO production (estimated through nitrites (NO(2) (-))), found between stage I (5.69 +/- 1.32 nmol/ml) and basal values (5.36 +/- 1.25 nmol/ml), was followed by the decrease in stage II (4.21 +/- 0.42 nmol/ml) and production lower than basal to the end of the test. Significant increase (p<0.05) in O(2) (-) values was found between stage I (4.18 +/- 0.77 nmol/ml) and resting values (4.01 +/- 0.69 nmol/ml), and at stages V (4.24 +/- 0.85 nmol/ml) and 1st phase of recovery (4.39 +/- 0.92 nmol/ml). CONCLUSION: The regression lines of NO(2) (-) and O(2) (-) crossed at the level of anaerobic threshold, suggesting that anaerobic threshold could be of a crucial importance not only in the anaerobic and aerobic metabolism but in mechanisms of signal transductions as well. Long-term exercise increases NO bioavailability, and there is positive correlation between NO bioavailability and maximal oxygen uptake (VO(2max)). PMID- 21633722 TI - Stability of the elbow joint: relevant anatomy and clinical implications of in vitro biomechanical studies. AB - The aim of this literature review is to describe the clinical anatomy of the elbow joint based on information from in vitro biomechanical studies. The clinical consequences of this literature review are described and recommendations are given for the treatment of elbow joint dislocation.The PubMed and EMBASE electronic databases and the Cochrane Central Register of Controlled Trials were searched. Studies were eligible for inclusion if they included observations of the anatomy and biomechanics of the elbow joint in human anatomic specimens.Numerous studies of the kinematics, kinesiology and anatomy of the elbow joint in human anatomic specimens yielded important and interesting implications for trauma and orthopaedic surgeons. PMID- 21633723 TI - Expression of vascular endothelial growth factor on chondrocytes increases with osteoarthritis - an animal experimental investigation. AB - OBJECTIVE: To evaluate the expression of VEGF by chondrocytes of hyaline cartilage during the course of osteoarthritis (OA). METHODS: In 12 white New Zealand rabbits the anterior cruciate ligament (ACL) was resected to create an anterior instability of the knee. In 12 control rabbits only a sham operation without resection of the ACL was done. Four animals of each group were killed at 3, 6, and 12 weeks. The load bearing area was evaluated histologically according to Mankin and by immunostaining for VEGF. RESULTS: In the experimental group, histological grades of OA showed a positive linear correlation with the time after surgery. Immunostaining showed an increased expression of VEGF in the control group after 3 weeks, which dropped to normal after 6 weeks. There was no difference in the progression of OA between control and experimental groups after 3 weeks, but a significant difference was seen after 6 (p=0,01) and 12 (p=0,05) weeks. A significant positive correlation between VEGF expression and the histological grade of OA was found (r = 0.767; p<0.01). CONCLUSIONS: An increase of VEGF expressing chondrocytes occurs during time course of OA. PMID- 21633724 TI - Effects of red grape juice consumption on high density lipoprotein-cholesterol, apolipoprotein AI, apolipoprotein B and homocysteine in healthy human volunteers. AB - It has suggested that grape juice consumption has lipid- lowering effect and it is associated with a decreased risk of heart disease. We aimed to evaluate the effects of red grape juice (RGj) consumption on high density lipoprotein cholesterol (HDL-C), apolipoprotein AI (apoAI), apolipoprotein B (apoB) and homocysteine (Hcy) levels in healthy human volunteers. Twenty six healthy and nonsmoking males, aged between 25-60 years, who were under no medication asked to consume 150 ml of RGj twice per day for one month. Serum HDL-C, apoAI, apoB and plasma Hcy levels were measured before and after one month RGj consumption. HDL-C levels after RGj consumption were significantly higher than the corresponding levels before the RGj consumption (41.44 +/- 4.50 and 44.37 +/- 4.30 mg/dl; P<0.0001). Also, apoB was significantly increased after RGj consumption (149.0 +/ 22.35 and 157.19 +/- 18.60 mg/dl; P<0.002). But apoAI levels were not changed significantly before and after of RGj consumption (154.27 +/- 21.55 and 155.35 +/ 21.07 mg/dl; P>0.05). Hcy levels were decreased after RGj consumption (7.70 +/- 2.80 and 6.20 +/- 2.30 umol/l; P<0.001). The present study demonstrates that RGj consumption can significantly increase serum HDL-C levels and decrease Hcy levels. These findings may have important implications for the prevention of atherosclerosis in healthy individuals. PMID- 21633725 TI - Anomaly of the deep lateral orbital wall in two cases. PMID- 21633727 TI - A current evaluation of the safety of angiotensin receptor blockers and direct renin inhibitors. AB - The safety of angiotensin II receptor blockers (ARBs) for the treatment of hypertension and cardiovascular and renal diseases has been well documented in numerous randomized clinical trials involving thousands of patients. However, recent concerns have surfaced about possible links between ARBs and increased risks of myocardial infarction and cancer. Less is known about the safety of the direct renin inhibitor aliskiren, which was approved as an antihypertensive in 2007. This article provides a detailed review of the safety of ARBs and aliskiren, with an emphasis on the risks of cancer and myocardial infarction associated with ARBs. Safety data were identified by searching PubMed and Food and Drug Administration (FDA) Web sites through April 2011. ARBs are generally well tolerated, with no known class-specific adverse events. The possibility of an increased risk of myocardial infarction associated with ARBs was suggested predominantly because the Valsartan Antihypertensive Long-Term Use Evaluation (VALUE) trial reported a statistically significant increase in the incidence of myocardial infarction with valsartan compared with amlodipine. However, no large scale, randomized clinical trials published after the VALUE study have shown a statistically significant increase in the incidence of myocardial infarction associated with ARBs compared with placebo or non-ARBs. Meta-analyses examining the risk of cancer associated with ARBs have produced conflicting results, most likely due to the inherent limitations of analyzing heterogeneous data and a lack of published cancer data. An ongoing safety investigation by the FDA has not concluded that ARBs increase the risk of cancer. Pooled safety results from clinical trials indicate that aliskiren is well tolerated, with a safety profile similar to that of placebo. ARBs and aliskiren are well tolerated in patients with hypertension and certain cardiovascular and renal conditions; their benefits outweigh possible safety concerns. PMID- 21633726 TI - Cardiovascular dysfunction in obesity and new diagnostic imaging techniques: the role of noninvasive image methods. AB - Obesity is a major public health problem affecting adults and children in both developed and developing countries. This condition often leads to metabolic syndrome, which increases the risk of cardiovascular disease. A large number of studies have been carried out to understand the pathogenesis of cardiovascular dysfunction in obese patients. Endothelial dysfunction plays a key role in the progression of atherosclerosis and the development of coronary artery disease, hypertension and congestive heart failure. Noninvasive methods in the field of cardiovascular imaging, such as measuring intima-media thickness, flow-mediated dilatation, tissue Doppler, and strain, and strain rate, constitute new tools for the early detection of cardiac and vascular dysfunction. These techniques will certainly enable a better evaluation of initial cardiovascular injury and allow the correct, timely management of obese patients. The present review summarizes the main aspects of cardiovascular dysfunction in obesity and discusses the application of recent noninvasive imaging methods for the early detection of cardiovascular alterations. PMID- 21633728 TI - Hepatocyte nuclear factor 4 alpha P2 promoter variants associate with insulin resistance. AB - This study aimed to investigate the associations of hepatocyte nuclear factor 4 (HNF4) alpha single nucleotide polymorphisms (SNPs) and haplotype with insulin resistance and metabolic syndrome parameters. Nine SNPs spanning the HNF4 alpha P2 promoter (rs4810424, rs1884613 and rs1884614) and coding region (rs2144908, rs6031551, rs6031552, rs1885088, rs1028583 and rs3818247) were genotyped in 160 subjects without diabetes or metabolic syndrome. The HNF4 alpha P2 promoter SNPs rs4810424, rs1884613 and rs1884614 were associated with insulin resistance (p = 0.017; 0.037; 0.024) and body mass index (BMI) (p = 0.03; 0.035; 0.039). The intron 1D SNP rs2144908 was associated with high-density lipoprotein cholesterol (HDLc) (p = 0.020) and the intron 9 SNP rs3818247 showed association with systolic (p = 0.02) and diastolic (p = 0.034) blood pressure. HNF4 alpha common haplotype CCCGTC associated with higher insulin resistance (p = 0.022), fasting blood glucose (FBG) (p = 0.035) and lower HDLc (p = 0.001). In conclusion, subjects with HNF4 alpha P2 variants and haplotypes have been shown to have a higher insulin resistance and are therefore at a higher risk for developing type 2 diabetes mellitus. PMID- 21633729 TI - Computational study of binding of epothilone A to beta-tubulin. AB - Understanding the interactions of epothilones with beta-tubulin is crucial for computer aided rational design of macrocyclic drugs based on epothilones and epothilone derivatives. Despite numerous structure-activity relationship investigations we still lack substantial knowledge about the binding mode of epothilones and their derivatives to beta-tubulin. In this work, we reevaluated the electron crystallography structure of epothilone A/beta-tubulin complex (PDB entry 1TVK) and proposed an alternative binding mode of epothilone A to beta tubulin that explains more experimental facts. PMID- 21633730 TI - Diagnosis and treatment difficulties in 18-year-old male patient with hereditary hemochromatosis, chronic hepatitis B, Gilbert syndrome and ulcerative colitis. AB - Among possible causes of chronic hepatitis in adolescents most common are infections, autoimmune disorders and metabolic diseases. Thus, diagnostic procedures should be multidirectional. This study reports diagnosis and treatment difficulties in an 18-year-old male patient with hereditary hemochromatosis (HH), ulcerative colitis (UC), chronic hepatitis B (CHB) and Gilbert syndrome. The presented case illustrates problems in diagnostics related to the presence of numerous disease conditions in one patient. It should be taken into consideration that these diseases coexisting in one patient can mutually affect their symptoms creating specific diagnostic difficulties. PMID- 21633731 TI - Electromagnetic and microwave-absorbing properties of magnetic nickel ferrite nanocrystals. AB - The electromagnetic and microwave absorbing properties of nickel ferrite nanocrystals were investigated for the first time. There were two frequencies corresponding to the maximum reflection loss in a wide thickness range from 3.0 to 5.0 mm, which may be bought by the nanosize effect and the good crystallization of the nanocrystals. PMID- 21633732 TI - Multicolored nanometre-resolution mapping of single protein-ligand binding complexes using far-field photostable optical nanoscopy (PHOTON). AB - Mapping of individual ligand molecules and their binding sites in single protein ligand complexes at nanometer resolution in real-time would enable probing their structures and functions in vitro and in vivo. In this study, we have developed far-field photostable optical nanoscopy (PHOTON) for mapping single ligand molecules (biotin) and their binding sites in individual protein-ligand complexes (streptavidin-biotin) with 1.2 nm spatial resolution and 100 ms temporal resolution. PHOTON includes one standard far-field optical microscope with a halogen-lamp illuminator; single-molecule-nanoparticle-optical-biosensors (SMNOBS) with exceptionally high quantum-yield (QY) of Rayleigh scattering and photostability (non-photobleaching, non-photoblinking) as imaging probes; and Multispectral Imaging System (MSIS) for spectral isolation of individual SMNOBS with 1 nm wavelength resolution. Intrinsic size- and shape- dependent localized surface-plasmon-resonance (LSPR) spectra of single SMNOBS provide multiple spectral (color) nanoprobes for sub-diffraction imaging, offering feasibility of probing of binding structures and functions of single protein-ligand complexes at nm (potentially achieving Angstrom) resolution in real-time. PMID- 21633733 TI - On molecular chirality within naturally occurring secondary organic aerosol particles from the central Amazon Basin. AB - In this perspectives article, we reflect upon the existence of chirality in atmospheric aerosol particles. We then show that organic particles collected at a field site in the central Amazon Basin under pristine background conditions during the wet and dry seasons consist of chiral secondary organic material. We show how the chiral response from the aerosol particles can be imaged directly without the need for sample dissolution, solvent extraction, or sample preconcentration. By comparing the chiral-response images with optical images, we show that chiral responses always originate from particles on the filter, but not all aerosol particles produce chiral signals. The intensity of the chiral signal produced by the size resolved particles strongly indicates the presence of chiral secondary organic material in the particle. Finally, we discuss the implications of our findings on chiral atmospheric aerosol particles in terms of climate related properties and source apportionment. PMID- 21633734 TI - The growth of single crystal silver wires at the nitrobenzene|water interface. AB - Single crystal silver wires can be grown at the nitrobenzene|water interface when silver ions dissolved in the aqueous phase are reduced by decamethyl ferrocene dissolved in the nitrobenzene phase. The successful growth of these wires depends on a number of experimental conditions, most prominently on the concentration ratio of reactants, nucleation rates, shape of formed nuclei, and wettability of nuclei. The size-time dependence can be modeled on the basis of microelectrode behavior of the silver nuclei and wire. AFM, SEM, light microscopy and single crystal X-ray diffraction has been applied to study the morphology of the silver nuclei and wires. PMID- 21633735 TI - Cooperative hydrogen bonding in trimers involving HCN and HBO. AB - A computational study of the cooperative effect of hydrogen bonding in linear trimers comprised of HCN and HBO molecules was undertaken at the MP2/6 311++G(2d,2p) level of theory. It was found that the third molecule leads to enhancement of the binding relative to that of the dimer species comprising these monomers. HBO is a better proton acceptor and a weaker proton donor than HCN. PMID- 21633736 TI - An infrared study of solid glycine in environments of astrophysical relevance. AB - The conversion from neutral to zwitterionic glycine is studied using infrared spectroscopy from the point of view of the interactions of this molecule with polar (water) and non-polar (CO(2), CH(4)) surroundings. Such environments could be found on astronomical or astrophysical matter. The samples are prepared by vapour-deposition on a cold substrate (25 K), and then heated up to sublimation temperatures of the co-deposited species. At 25 K, the neutral species is favoured over the zwitterionic form in non-polar environments, whereas for pure glycine, or in glycine/water mixtures, the dominant species is the latter. The conversion is easily followed by the weakening of two infrared bands in the mid IR region, associated to the neutral structure. Theoretical calculations are performed on crystalline glycine and on molecular glycine, both isolated and surrounded by water. Spectra predicted from these calculations are in reasonable agreement with the experimental spectra, and provide a basis to the assignments. Different spectral features are suggested as probes for the presence of glycine in astrophysical media, depending on its form (neutral or zwitterionic), their temperature and composition. PMID- 21633744 TI - O-O bond activation in H2O2 and (CH3)3C-OOH mediated by [Ni(cyclam)(CH3CN)2](ClO4)2: different mechanisms to form the same Ni(III) product? AB - Reaction of [Ni(II)(cyclam)(CH(3)CN)(2)](ClO(4))(2) (1) with tert butylhydroperoxide (TBHP) or H(2)O(2), in acidic media results in a formation of [Ni(III)(cyclam)(CH(3)CN)(2)](3+) species (2), the nature of which is characterized by UV-vis, EPR and XPS. The formation rate of 2 is much higher when H(2)O(2) is used as oxidant. In absence of acid, TBHP reacts with 1 generating the same Ni(III) species but, in contrast, no reaction is observed between H(2)O(2) and 1. Addition of cis-stilbene as an oxidable substrate quenches the formation of Ni(III) for reaction with H(2)O(2) only. Overall, these observations reveal a different reaction mechanism when reacting H(2)O(2) with 1 than when reacting the same metal complex with TBHP, despite the fact that the final product is the same. The proposed pathways arising from these observations consist in a homolytic O-O cleavage for the reaction of 1 with TBHP in CH(3)CN, but a proton assisted heterolysis for the O-O activation in H(2)O(2). Density functional calculations (B3LYP and OPBE) on the thermodynamic feasibility of the two reaction processes support the proposed mechanisms, since the O-O homolysis is strongly disfavored when H(2)O(2) is used as reactant. PMID- 21633745 TI - Label-free, regenerative and sensitive surface plasmon resonance and electrochemical aptasensors based on graphene. AB - Label-free, regenerative and sensitive surface plasmon resonance (SPR) and electrochemical aptasensors based on graphene for the detection of alpha-thrombin have been reported, which propose a new, simple way for protein recognition with high sensitivity and selectivity. PMID- 21633746 TI - A reversible fluorescence nanoswitch based on bifunctional reduced graphene oxide: use for detection of Hg2+ and molecular logic gate operation. AB - Herein, we demonstrate the first use of a reduced graphene oxide (rGO)-organic dye nanoswitch for the label-free, sensitive and selective detection of Hg(2+) using bifunctional rGO as an effective nanoquencher and highly selective nanosorbent. Moreover, a reversible on-off INHIBIT rGO logic gate based on a cysteine-Hg(2+) system has also been designed. PMID- 21633747 TI - Large-scale synthesis of uniform silver orthophosphate colloidal nanocrystals exhibiting high visible light photocatalytic activity. AB - Silver orthophosphate nanocrystals with controlled particle size have been synthesized using a simple, reproducible and easily scaled up route based on the reaction between silver ions, oleylamine and phosphoric acid. The obtained nanocrystals are highly uniform in size and exhibit high visible light activity for the photodecomposition of organic compounds. PMID- 21633748 TI - Total synthesis of (+/-)-chamobtusin A. AB - The total synthesis of chamobtusin A, the first diterpenoid alkaloid isolated from the whole Pinales, is described. Key features of the synthesis include a stereoselective intramolecular Michael addition to install a key stereocenter and an oxidative manipulation to prepare a 2H-pyrrole ring. PMID- 21633749 TI - Stereoselective synthesis of tetracyclic indolines via gold-catalyzed cascade cyclization reactions. AB - A reliable synthetic route to fused polycyclic indolines is documented by the development of a stereoselective gold catalyzed cascade cyclization of indole propargylic alcohols. PMID- 21633750 TI - Manganese(III)-mediated phosphinoyl radical reactions for stereoselective synthesis of phosphinoylated tetrahydronaphthalenes. AB - Reactions of diphenylphosphinoyl radicals with 5-aryl-2-pentenoates or beta phenylethylene styrenes generate trans-substituted tetrahydronaphthalenes through a cascade reaction sequence. PMID- 21633751 TI - The protonation state of small carboxylic acids at the water surface from photoelectron spectroscopy. AB - We report highly surface sensitive core-level photoelectron spectra of small carboxylic acids (formic, acetic and butyric acid) and their respective carboxylate conjugate base forms (formate, acetate and butyrate) in aqueous solution. The relative surface propensity of the carboxylic acids and carboxylates is obtained by monitoring their respective C1s signal intensities from a solution in which their bulk concentrations are equal. All the acids are found to be enriched at the surface relative to the corresponding carboxylates. By monitoring the PE signals of acetic acid and acetate as a function of total concentration, we find that the protonation of acetic acid is nearly complete in the interface layer. This is in agreement with literature surface tension data, from which it is inferred that the acids are enriched at the surface while (sodium) formate and acetate, but not butyrate, are depleted. For butyric acid, we conclude that the carboxylate form co-exists with the acid in the interface layer. The free energy cost of replacing an adsorbed butyric acid molecule with a butyrate ion at 1.0 M concentration is estimated to be >5 kJ mol(-1). By comparing concentration dependent surface excess data with the evolution of the corresponding photoemission signals it is furthermore possible to draw conclusions about how the distribution of molecules that contribute to the excess is altered with bulk concentration. PMID- 21633752 TI - Enhanced electrochemiluminescence quenching of CdS:Mn nanocrystals by CdTe QDs doped silica nanoparticles for ultrasensitive detection of thrombin. AB - This work reports an aptasensor for ultrasensitive detection of thrombin based on remarkably efficient energy-transfer induced electrochemiluminescence (ECL) quenching from CdS:Mn nanocrystals (NCs) film to CdTe QDs-doped silica nanoparticles (CdTe/SiO(2) NPs). CdTe/SiO(2) NPs were synthesized via the Stober method and showed black bodies' strong absorption in a wide spectral range without excitonic emission, which made them excellent ECL quenchers. Within the effective distance of energy scavenging, the ECL quenching efficiency was dependent on the number of CdTe QDs doped into the silica NPs. Using ca. 200 CdTe QDs doped silica NPs on average of 40 nm in diameter as ECL quenching labels, attomolar detection of thrombin was successfully realized. The protein detection involves a competition binding event, based on thrombin replacing CdTe/SiO(2) NPs labeled probing DNA which is hybridized with capturing aptamer immobilized on a CdS:Mn NCs film modified glassy carbon electrode surface by specific aptamer protein affinity interactions. It results in the displacement of ECL quenching labels from CdS:Mn NCs film and concomitant ECL signal recovery. Owing to the high-content CdTe QDs in silica NP, the increment of ECL intensity (DeltaI(ECL)) and the concentration of thrombin showed a double logarithmic linear correlation in the range of 5.0 aM~5.0 fM with a detection limit of 1aM. And, the aptasensor hardly responded to antibody, bovine serum albumin (BSA), haemoglobin (Hb) and lysozyme, showing good detection selectivity for thrombin. This long-distance energy scavenging could have a promising application perspective in the detection of biological recognition events on a molecular level. PMID- 21633753 TI - Phase behavior of PCBM blends with different conjugated polymers. AB - In this work the phase behavior of [6,6]-phenyl C(61)-butyric acid methyl ester (PCBM) blends with different poly(phenylene vinylene) (PPV) samples is investigated by means of standard and modulated temperature differential scanning calorimetry (DSC and MTDSC) and rapid heat-cool calorimetry (RHC). The PPV conjugated polymers include poly(2-methoxy-5-(3',7'-dimethyloctyloxy)-1,4 phenylene vinylene) (MDMO-PPV), High T(g)-PPV which is a copolymer, and poly((2 methoxy-5-phenethoxy)-1,4-phenylene vinylene) (MPE-PPV). Comparisons of these PPV:PCBM blends with regioregular poly(3-hexyl thiophene) (P3HT):PCBM blends are made to see the different component miscibilities among different blends. The occurrence of liquid-liquid phase separation in the molten state of MDMO-PPV:PCBM and High T(g)-PPV:PCBM blends is indicated by the coexistence of double glass transitions for blends with a PCBM weight fraction of around 80 wt%. This is in contrast to the P3HT:PCBM blends where no phase separation is observed. Due to its high cooling rate (about 2000 K min(-1)), RHC proves to be a useful tool to investigate the phase separation in PPV:PCBM blends through the glass transition of these crystallizable blends. P3HT is found to have much higher thermal stability than the PPV samples. PMID- 21633754 TI - Anaesthetic management of bronchopleural fistula in a patient with myasthenia gravis. AB - Patients with bronchopleural fistula present with specific airway management and ventilatory concerns, which makes the anaesthetic management of these patients challenging. Myasthenia gravis is another condition requiring specific anaesthetic management, with possible unpredictable delays in recovery. A combination of both these conditions in a patient makes management even more difficult. Our patient with myasthenia gravis underwent repair of the bronchopleural fistula, during which a multimodal approach to intraoperative and postoperative analgesia was adopted. Positive pressure ventilation was started only after we confirmed the isolation of the lung. PMID- 21633755 TI - Polymyositis in association with polycythaemia vera. AB - Inflammatory myopathies are reported to be associated with various malignancies. This association is more commonly observed in cases of dermatomyosis and less frequently in polymyositis. Malignancies commonly reported in association with inflammatory myopathies include non-Hodgkin's lymphoma and ovarian, lung and gastric carcinomas, as well as nasopharyngeal malignancies. Polycythaemia vera as a cause of polymyositis is unknown. We report a case of polycythaemia vera in a female patient who developed polymyositis three years later. PMID- 21633756 TI - Pericardiophrenic artery embolisation for control of massive haemoptysis. AB - Non-bronchial systemic arteries, apart from normal and anomalous bronchial arteries, may be a source of massive haemoptysis in a chronically inflamed lung via transpleural anastomoses. Transcatheter embolisation is an established therapeutic method of choice in the management of massive haemoptysis. We report embolisation of a hypertrophied pleural branch of the pericardiophrenic artery for the management of massive haemoptysis in a 61-year-old woman. Initial computed tomography chest imaging showed peribronchial thickening and subpleural scarring in the lingula lobe, with ground-glass changes secondary to haemoptysis. Angiography demonstrated a hypertrophied branch of the left pericardiophrenic artery supplying an abnormal bunch of vessels in the lingula and anastomosing with the homolateral inferior phrenic artery. This was successfully embolised with gel foam. The left internal thoracic artery was later embolised in order to control the repeat haemoptysis. A brief anatomical review of the source of massive haemoptysis, anatomy of the internal thoracic and pericardiophrenic arteries and the clinical implications are discussed. PMID- 21633757 TI - Occupational contact dermatitis in manual cloud seeding operations. AB - This is a case report on irritant contact dermatitis secondary to calcium oxide exposure during manual cloud seeding operations. A less hazardous substitute such as sodium chloride should be considered wherever possible. Cloud seeding operations are briefly discussed in this report, and the impact of calcium oxide exposure as an occupational hazard is elaborated. PMID- 21633758 TI - Acute fibrinous and organising pneumonia presenting as complete lung consolidation. AB - Acute fibrinous and organising pneumonia (AFOP) is an unusual histopathological pattern of acute lung injury. The clinical manifestations, course and treatment of AFOP have yet to be characterised. All reported cases so far have described bilateral diffuse lung involvement radiologically. We report a case of an adolescent girl who presented with acute hypoxaemic respiratory failure with unilateral complete lung consolidation. She was initially diagnosed with severe community-acquired pneumonia. A computed tomography-guided percutaneous transthoracic trucut biopsy of the left lung revealed the classical histopathological pattern typically observed in AFOP. The patient responded well to treatment involving steroids. The uniqueness of such a presentation in AFOP prompted us to report this case. PMID- 21633759 TI - Type VI choledochal cyst revisited. AB - Choledochal cysts are rare cystic dilatations of the bile ducts, which are commonly found in children. Choledochal cysts in adults are uncommon, and cystic dilatation of the cystic duct (type VI choledochal cyst) is a unique occurrence. To date, specific pathogenic and diagnostic criteria for describing type VI cysts have been lacking. This report attempts to lay down the specific diagnostic and management criteria of this uncommon entity, with reviews of all available cases reported in the literature. PMID- 21633760 TI - Abdominal foreign body: late presentation as a rectus sheath abscess. AB - Intra-abdominal ingested foreign bodies are usually an incidental finding, typically encountered in mentally challenged patients. We present the case of a 65-year-old mentally sound woman who presented with recurrent abdominal pain and a lump in the hypogastrium. Evaluation revealed a rectus sheath abscess extending to the peritoneum, with a foreign body in situ. On enquiry, the patient revealed that she had accidentally ingested a tailoring needle 17 years ago. This case illustrates an unusual presentation of an ingested foreign body as a rectus sheath abscess after a long duration. PMID- 21633761 TI - Tracheobronchial injuries following endotracheal intubation. AB - We report two rare cases of tracheobronchial injury (TBI) following endotracheal intubation. Both intubations were easy and performed by experienced anaesthetists. The injuries for both cases were possibly caused by the tip of an endotracheal tube. A 27-year-old woman regurgitated copiously during induction of general anaesthesia for a Caesarean section. She had a full-thickness TBI 2 cm above the carina, which was repaired through an open thoracotomy. A 68-year-old woman undergoing left mastectomy was intubated with a 7.5-mm endotracheal tube for general anaesthesia. 24 hours after extubation, the patient developed widespread subcutaneous emphysema. There was no associated respiratory distress, and the patient was treated conservatively. The fragile tracheal tissue associated with pregnancy and old age was possibly a contributing factor for injury. Both patients recovered well. Surgical and non-surgical methods can be safely used after careful consideration of the patient's clinical, radiological and endoscopic findings. PMID- 21633763 TI - Sigmund Freud (1856-1939): father of psychoanalysis. PMID- 21633764 TI - Aviation Medicine: global historical perspectives and the development of Aviation Medicine alongside the growth of Singapore's aviation landscape. AB - Aviation Medicine traces its roots to high altitude physiology more than 400 years ago. Since then, great strides have been made in this medical specialty, initially catalysed by the need to reduce pilot medical attrition during the World Wars, and more recently, fuelled by the explosive growth in globalised commercial air travel. This paper traces the historical milestones in Aviation Medicine, and maps its development in Singapore since the 1960s. Advancements in military aviation platforms and technology as well as the establishment of Singapore as an international aviation hub have propelled Aviation Medicine in Singapore to the forefront of many domains. These span Aviation Physiology training, selection medical standards, performance maximisation, as well as crew and passenger protection against communicable diseases arising from air travel. The year 2011 marks the centennial milestone of the first manned flight in Singapore, paving the way for further growth of Aviation Medicine as a mature specialty in Singapore. PMID- 21633765 TI - Variations of atrioventricular block. AB - Atrioventricular (AV) block comprises a spectrum of cardiac conduction delays with varying clinical presentations. It is commonly encountered in both hospital as well as ambulatory settings, and recognition of the type of AV conduction delay is essential for appropriate subsequent management. The electrocardiogram is a key tool for identification of patients with AV conduction delays. Contrasting management strategies should be employed for differing levels of conduction block. PMID- 21633766 TI - PlasmaKineticTM (bipolar) transurethral resection of prostate: a prospective trial to study pathological artefacts, surgical parameters and clinical outcomes. AB - INTRODUCTION: The aims of the study were to compare the degree of cautery artefacts in prostatic chips between monopolar and PlasmaKineticTM transurethral resection of prostate (TURP), and to determine if there is any difference in the intraoperative and post surgical parameters between them. METHODS: After institutional review board approval, patients were prospectively enrolled to undergo PlasmaKineticTM TURP. Their parameters were compared with those of the historical monopolar TURP controls. All histological specimens were reviewed by a single senior pathologist. RESULTS: 46 patients were recruited to undergo PlasmaKineticTM TURP. The resection time was significantly longer for the bipolar group compared to the monopolar group (50.2 versus 36.7 min, p-value is 0.001). The speed of resection (resection weight/time) was lower for the bipolar group (0.45 versus 0.56 g/min, p-value is 0.017). More irrigant was used for the bipolar group (21.2 versus 15.6 litres, p-value is 0.001) intraoperatively. There was no statistically significant difference in terms of intraoperative drop in haemoglobin and serum sodium change between the two groups. There seems to be a lesser degree of cautery artefacts in the PlasmaKineticTM group than the monopolar group (42.17 versus 45.07 microns); however, this was not statistically significant (p-value is 0.452). CONCLUSION: Bipolar TURP seems to result in a lesser degree of cautery artefacts when compared to conventional monopolar TURP, albeit statistically insignificant, compared to monopolar TURP. TURP also resulted in a longer resection time and increased irrigant use, but no difference in blood loss and serum sodium levels. PMID- 21633767 TI - Comparison of RIPASA and Alvarado scores for the diagnosis of acute appendicitis. AB - INTRODUCTION: The accuracy of the Alvarado score in diagnosing acute appendicitis in an Asian population has been disappointingly low. We prospectively compared the RIPASA score with the Alvarado score for the diagnosis of acute appendicitis. METHODS: 200 consecutive patients who presented to the Accident and Emergency Department with right iliac fossa pain were recruited in the study. Both the RIPASA and Alvarado scores were derived, but decisions for appendicectomy were based on clinical judgement. Receiver operating curve (ROC), sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) for both scoring systems were calculated. RESULTS: Only 192 out of the 200 patients who satisfied the inclusion and exclusion criteria were included in the analysis. At the optimal cut-off threshold score of 7.5 derived from the ROC, the sensitivity, specificity, PPV, NPV and diagnostic accuracy of the RIPASA score were 98.0 percent, 81.3 percent, 85.3 percent, 97.4 percent and 91.8 percent, respectively. At the cut-off threshold score of 7.0 for the Alvarado score, the sensitivity, specificity, PPV, NPV and diagnostic accuracy were 68.3 percent, 87.9 percent, 86.3 percent, 71.4 percent and 86.5 percent, respectively. The RIPASA score correctly classified 98 percent of all patients confirmed with histological acute appendicitis to the high-probability group (RIPASA score greater than 7.5) compared with 68.3 percent with the Alvarado score (Alvarado score greater than 7.0; p-value less than 0.0001). CONCLUSION: The RIPASA score at a cut-off threshold total score of 7.5 is a better diagnostic scoring system than the Alvarado score for the diagnosis of acute appendicitis in our local setting. PMID- 21633768 TI - Melioidosis of the extremities in Brunei Darussalam. AB - INTRODUCTION: Melioidosis caused by Burkholderia pseudomallei is an infectious disease endemic to Southeast Asia and northern Australia. It has a broad spectrum of clinical manifestations and high mortality, and can mimic other infectious diseases. The aim of this study was to review cases of melioidosis of the extremities in Brunei Darussalam. METHODS: Culture-positive cases for Burkholderia pseudomallei in Raja Isteri Pengiran Anak Saleha Hospital were identified from records in the Microbiology Department. The case notes were reviewed to identify patients who were treated for problems affecting the extremities. 14 (13 males and one female) out of 48 patients were identified. RESULTS: The median age of the patients was 45 (range 14-55) years. Septicaemia was the most common presenting feature in 11 patients. Multisystem involvement was noted in eight patients, diabetes mellitus in nine patients and other risk factors in two patients. Blood culture was positive in ten patients and pus culture in 11 patients. The presentations noted were cellulitis of the limbs, abscess, osteomyelitis (three patients each) and septic arthritis (five patients). Orthopaedic intervention (joint washout/incision and drainage/curettage) was required in 11 patients. The median hospital stay was 27.5 (range 13-63) days; two patients required admission to intensive care. No mortality was reported. CONCLUSION: Melioidosis of the extremities is not uncommon in Brunei Darussalam. It is associated with significant morbidity, and a large number of patients require surgical intervention. Thus, a high index of suspicion is required for early diagnosis and institution of appropriate antibiotic therapy. PMID- 21633769 TI - Bacteriology of deep neck abscesses: a retrospective review of 96 consecutive cases. AB - INTRODUCTION: This study aimed to review the microbiology of deep neck abscesses and identify the factors that influence their occurrence. METHODS: A retrospective chart review was done of patients diagnosed with deep neck abscesses at the Department of Otorhinolaryngology, Tan Tock Seng Hospital, Singapore between 2004 and 2009. The data of 131 deep neck abscess patients were reviewed, and those with positive pus culture were included in the study. Logistic regression was applied to analyse and compare the incidence of common organisms in various conditions (age, gender, aetiology and effects of diabetes mellitus). RESULTS: Of the 96 patients recruited, 18 had polymicrobial cultures. The leading pathogens cultured were Klebsiella (K.) pneumoniae (27.1 percent), Streptococcus milleri group (SMG) bacteria (21.9 percent) and anaerobic bacteria not otherwise specified (NOS) (20.8 percent). K. pneumoniae (50.0 percent) was over-represented in the diabetic group. SMG bacteria (68.8 percent) and anaerobic bacteria-NOS (43.8 percent) were most commonly isolated in patients with odontogenic infections. K. pneumoniae was found more commonly among female patients (39.3 percent). The distribution of the three leading pathogens between patients aged below 50 years and those 50 years and above was comparable. K. pneumoniae was the commonest organism cultured in parapharyngeal space abscesses, while the submandibular space and parotid space most commonly isolated SMG bacteria and Staphylococcus aureus, respectively. CONCLUSION: Broad-spectrum antibiotics are recommended for treating deep neck abscesses. Empirical antibiotic coverage against K. pneumoniae infection in diabetic patients, and SMG and anaerobic bacteria in patients with an odontogenic infection, is advocated. Routine antibiotic coverage against Gram-negative bacteria is not paramount. PMID- 21633770 TI - A randomised controlled trial of glutamine-enriched neonatal parenteral nutrition in Malaysia. AB - INTRODUCTION: The addition of glutamine to parenteral nutrition (PN) in neonates has not shown significant benefits as compared to adults thus far. This study aimed to determine the potential benefits of the addition of glutamine to neonatal PN in a tertiary hospital in a middle-income country. METHODS: This was a double-blinded randomised controlled trial. Babies who were admitted to the neonatal intensive care unit (NICU) and who required PN were eligible for inclusion in the study. The subjects were randomised to receive either glutamine added PN (intervention) or standard PN (control). The most important outcomes included time to full enteral nutrition, incidence of sepsis and necrotising enterocolitis (NEC), clinical or culture-proven sepsis. RESULTS: Out of 270 subjects, 132 were randomised to the intervention group and 138, to the control group. Baseline data were comparable in both groups. The median time taken to reach full enteral nutrition was similar for both intervention and control groups (six days in each group, p-value is 0.52). The incidences of NEC, clinical sepsis and culture-proven sepsis did not differ significantly in the intervention and control groups (5.8 vs. 7.1 percent, p-value is 0.68; 15.7 percent vs. 10.2 percent, p-value is 0.21 and 16.5 percent vs. 15.7 percent, p-value is 0.38, respectively). Other outcomes such as duration of ventilation, duration of NICU stay and a subgroup analysis for preterm and term babies also showed no statistically significant differences. CONCLUSION: Addition of glutamine to neonatal PN was not shown to improve outcome. PMID- 21633771 TI - Isolation of the first three cases of Clostridium difficile polymerase chain reaction ribotype 027 in Singapore. AB - INTRODUCTION: The incidence of Clostridium (C.) difficile infection (CDI) was on the rise from 2001 to 2006 in Singapore. Recent unpublished data suggests that its incidence had remained stable or decreased in most local public hospitals between 2006 and 2010. It is, however, not known if the polymerase chain reaction (PCR) ribotype 027 strains have been circulating, although reports suggest that this strain is emerging in Asia, with the first cases reported from Japan in 2007, as well as in Hong Kong and Australia in 2009. We initiated a culture-based surveillance to detect this epidemic strain in Singapore. METHODS: From September 2008 to December 2009, all non-duplicate toxin-positive stool samples from the three largest public hospitals in Singapore were collected for culture and further analysis. RESULTS: Out of the 366 samples collected, 272 viable isolates were cultured. Of these, 240 tested toxin-positive and ten tested positive for the binary toxin gene; 35 different PCR ribotypes were found. Three isolates that tested positive for binary toxin contained the same PCR ribotyping pattern as the C. difficile 027 control strain. All three had the 18-bp deletion and single nucleotide tcdC deletion at position 117. Susceptibility testing was performed, demonstrating susceptibility to erythromycin and moxifloxacin. CONCLUSION: We report the first three isolates of C. difficile 027 from Singapore. However, their susceptibility patterns are more consistent with the historical 027 strains. Rising CDI incidence may not be associated with the emergence of the epidemic 027 strain at this time. PMID- 21633772 TI - Human bocavirus in Jordan: prevalence and clinical symptoms in hospitalised paediatric patients and molecular virus characterisation. AB - INTRODUCTION: This study was aimed at investigating the prevalence of human bocavirus (HBoV) among Jordanian children hospitalised with lower respiratory tract infection (LRTI) as well as the clinical feature associated with HBoV infection, the seasonal distribution of HBoV and the DNA sequencing of HBoV positive samples. METHODS: A total of 220 nasopharyngeal aspirates were collected from children below 13 years of age who were hospitalised with LRTI in order to detect the presence of HBoV using real-time polymerase chain reaction assay and direct HBoV sequencing. RESULTS: HBoV was detected in 20 (9.1 percent) patients, whose median age was four (range 0.8-12) months. Children under the age of 12 months were more susceptible to HBoV infection (p-value is 0.016). The main clinical diagnoses of patients infected with HBoV were bronchopneumonia (35 percent) and bronchiolitis (30 percent). Coughing (100 percent), wheezing (82.7 percent) and fever (68.2 percent) were the most prominent symptoms in infected patients. HBoV infections were seasonal; increasing in cooler months, diminishing in the summer and peaking in March (45 percent). Direct DNA sequencing revealed that three out of 20 (15 percent) specimens were identical to Stockholm 1 and 2 isolates, and single base pair substitution (A to T) at codon 92 was found in 17 out of the 20 (85 percent) specimens that were positive for HBoV, resulting in a threonine-to-serine substitution. CONCLUSION: More attention should be given to diagnosing HBoV in patients with LRTI using molecular techniques. PMID- 21633773 TI - Outcome of adult patients with acute lymphoblastic leukaemia receiving the MRC UKALL XII protocol: a tertiary care centre experience. AB - INTRODUCTION: Acute lymphoblastic leukaemia (ALL) is a heterogeneous group of lymphoid neoplasm resulting from the proliferation of malignant lymphoid cells. We aimed to study the outcome of adult patients with ALL receiving the Medical Research Council UKALL XII protocol. METHODS: This was a retrospective study conducted at Aga Khan University Hospital from January 2001 to December 2008. The medical records of all adult patients were reviewed and analysed for clinical, morphological and immunological features at presentation and impact on treatment outcomes. Multivariate analysis and survival studies were performed using Kaplan Meier statistics. RESULTS: The total number of patients was 54, with a male to female ratio of 3.4:1 and a median age of 28 years. Common presenting symptoms were fever (n is 49) and bleeding (n is 14). 38 patients had haemoglobin less than 10 gms/dl, 21 had white blood cell (WBC) count of 50 * 10E9/L or more, and 35 had lactate dehyrogenase more than 1,000 IU. Morphologically, FAB-L2 was the commonest subtype, with 38 patients with B-ALL and eight with T-ALL. Multivariate analysis showed that age above 30 years, male gender, WBC count above 50 * 10E9/L and T-ALL subtype were independent risk factors for poor survival. 46 (85 percent) patients achieved complete remission. The median survival was 12.3 months. At the end of five years, 16 patients were alive, two were alive with disease and 14 were in complete remission. CONCLUSION: Overall survival and relapse rates in our study were comparable to those reported internationally. PMID- 21633774 TI - Comments on: Prolonged cough presenting with diagnostic difficulty: a study of aetiological and clinical outcomes. PMID- 21633777 TI - Ovarian hyperstimulation syndrome. PMID- 21633779 TI - Haematological parameters in patients with non-alcoholic fatty liver disease and alcoholic liver disease: concern on findings. PMID- 21633780 TI - Clinics in diagnostic imaging (135). Cystic renal cell carcinoma. AB - A 45-year-old man presented with right flank pain and haematuria for one month. Computed tomography showed a large, well-circumscribed exophytic complex cystic mass with enhanced, irregular thickened walls arising from the upper pole of the right kidney, which was associated with bilateral renal stones. Partial right nephrectomy with removal of the right renal stones was performed. Histopathology revealed papillary cell carcinoma confined to the kidney. The patient made good postoperative recovery. The Bosniak classification system of renal cystic lesions and cystic renal cell carcinoma are discussed. Various cases of renal cystic lesions and cystic renal cell carcinoma are shown. PMID- 21633782 TI - Fibroblast growth factor 23 (FGF23) and alpha-klotho stimulate osteoblastic MC3T3.E1 cell proliferation and inhibit mineralization. AB - Elevated serum levels of the phosphate-regulating hormone fibroblast growth factor 23 (FGF23) are found in patients with phosphate wasting diseases and chronic kidney disease-mineral and bone disorder (CKD-MBD). These diseases are associated with rickets and renal osteodystrophy, respectively. FGF23 is secreted from osteoblastic cells and signals through FGFRs, membrane coreceptor alpha Klotho (Klotho), and, possibly, a circulating form of Klotho. Despite the absence of detectable Klotho on osteoblastic cells, studies have suggested that forced FGF23 expression in osteoblasts inhibited mineralization. Thus, we examined the effects of exogenously applied FGF23 on osteoblastic MC3T3.E1 cell proliferation and differentiation, with and without soluble Klotho. MC3T3.E1 cells were cultured in osteoblast differentiation medium, supplemented with FGF23 (0.1-1,000 ng/mL), Klotho (50 ng/mL), the combination FGF23 + Klotho, and FGF2 (100 ng/mL) as a control. Neither FGF23 nor Klotho exposure affected proliferation of day 4 growth phase cells or mineralization of day 14 cultures. In contrast, FGF23 + Klotho resulted in inhibition of mineralization and osteoblast activity markers at day 14, and a slight, reproducible induction of proliferation. Inhibition of FGFR1, but not FGFR2 or FGFR3, completely restored FGF23 + Klotho-induced inhibition of alkaline phosphatase (ALP) activity at day 7. ALP activity was partially restored by the MAPK inhibitor U0126 but not inhibitors p38 and P13K. Thus, soluble Klotho enables FGF23 signaling in MC3T3.E1 cells, likely through FGFR 1(IIIc). Elevated FGF23 actions, in part, appear to parallel FGF2 with lower potency. In addition to affecting bone via indirect phosphate wasting pathways, supraphysiological FGF23 and soluble Klotho may directly impact bone in diseases with elevated FGF23 levels. PMID- 21633783 TI - Oxymatrine ameliorates L-arginine-induced acute pancreatitis in rats. AB - The aim of this study was to determine whether oxymatrine has a protective effect against acute pancreatitis (AP) in a rat model of L-arginine-induced AP. AP was induced by two intraperitoneal injections of L-arginine (250 mg/100 g) at a 1-h interval. Oxymatrine (50 mg/kg) was administered every 6 h after the induction of AP. Oxymatrine significantly reduced the plasma amylase, D-lactic acid and tumor necrosis factor alpha concentration, serum diamine oxidase and lipase activity, and pancreatic myeloperoxidase activity, which were increased in AP rats (P < 0.05). In addition, the pancreatic CD45 expression and the expression of claudin 1, but not zonula occludens-1 (ZO-1) and occludin, in the intestinal tissues were significantly reduced after the induction of AP. However, oxymatrine increased the expression of claudin-1 and CD45, but did not alter the expression of ZO-1 and occludin. In conclusion, our results demonstrated that oxymatrine is potentially capably of protecting against L-arginine-induced AP and attenuating AP-associated intestinal barrier injury by up-regulation of claudin-1. PMID- 21633785 TI - The evolving role of targeted therapy in early-stage and locally advanced non small cell lung cancer. AB - Many of the leading developments in management of non-small cell lung cancer (NSCLC) have been provided by the integration of specific targeted therapies either in combination with a backbone of standard chemotherapy or as a single agent. Agents that inhibit a specific pathway, such as that triggered by the activity of the epidermal growth factor receptor, or a regulatory process like angiogenesis, have made it possible to markedly increase response rates and extend survival, sometimes dramatically, along with a favorable therapeutic index. However, these novel therapies have established clinical benefit thus for only in the setting of incurable, advanced NSCLC. The value of these strategies in not only extending survival but potentially improving the cure rate when added to, or substituting for, conventional chemotherapy in the setting of early-stage resectable NSCLC or locally advanced NSCLC remains to be determined. A wide range of clinical trials for these settings have been pursued, with several pivotal studies still ongoing, and will be reviewed for their potential to redefine our current standards of care for potentially curable NSCLC through the integration of targeted therapies. PMID- 21633784 TI - Systematic therapy for unresectable or metastatic soft-tissue sarcomas: past, present, and future. AB - Unresectable or metastatic disease occurs in 40% to 60% of soft-tissue sarcoma (STS) patients and portends a poor prognosis. For decades, doxorubicin has formed the backbone of systemic treatment, with response rates of approximately 26%. Patients progressing following first-line therapy were left with few proven options. No other cytotoxic chemotherapy agent or combination has demonstrated superiority to doxorubicin. Advances in targeted therapy of STS have been hindered by STS heterogeneity and poorly understood disease biology. Despite challenges, progress has been made in specific STS subtypes. Here, we highlight the challenges, progress, and lessons learned from STS trials published in the last 20 to 25 years. PMID- 21633787 TI - Polymorphisms in the promoter region of the CASP8 gene are not associated with non-Hodgkin's lymphoma in Chinese patients. AB - Caspase-8 (CASP8) involved in apoptosis plays an important role in mediating the normal regulation of cell proliferation, differentiation, inflammation, and homeostasis of multicellular organisms. Genetic polymorphisms, rs3834129 ( /CTTACT) and rs3769821 (T/C), in the promoter region of different CASP8 transcripts, were reported to be associated with genetic susceptibility of multiple cancers and non-Hodgkin's lymphoma (NHL), respectively. To investigate whether these two genetic variants, together with rs113686495 (-/CTGTCATT) which is 50 bp downstream of rs3769821, were associated with NHL in Chinese patients, we genotyped two cohorts of case and control samples from Kunming (case n = 64, control n = 133) and Shanghai (case n = 75, control n = 107). Luciferase assays were further performed to characterize the potential role of different alleles in the promoter region of the CASP8 gene. In contrast to previous studies, we found no difference regarding the genotypes and haplotypes of rs3834129, rs3769821, and rs113686495 between the case and control samples. Luciferase assays of the promoter regions harboring different alleles of these three variants also showed no difference. Our negative results gave no support for an active role for these genetic variants in conferring NHL in Chinese patients. PMID- 21633786 TI - Beta-3 adrenoceptors as new therapeutic targets for cardiovascular pathologies. AB - Catecholamines play a key role in the regulation of cardiovascular function, classically through beta(1/2)-adrenoreceptors (AR) activation. After beta(3)-AR cloning in the late 1980s, convincing evidence for beta(3)-AR expression and function in cardiovascular tissues recently initiated a reexamination of their involvement in the pathophysiology of cardiovascular diseases. Their upregulation in diseased cardiovascular tissues and resistance to desensitization suggest they may be attractive therapeutic targets. They may substitute for inoperant beta(1/2)-AR to mediate vasodilation in diabetic or atherosclerotic vessels. In cardiac ventricle, their contractile effects are functionally antipathetic to those of beta(1/2)-AR; in normal heart, beta(3)-ARs may mediate a moderate negative inotropic effect, but in heart failure, it may protect against adverse effects of excessive catecholamine stimulation by action on excitation contraction coupling, electrophysiology, or remodelling. Thus, prospective studies in animals and patients at different stages of heart failure should lead to identify the best therapeutic window to use beta(3)-AR agonists and/or antagonists. PMID- 21633788 TI - QSAR model toward the rational design of new agrochemical fungicides with a defined resistance risk using substructural descriptors. AB - The increasing resistance of several phytopathogenic fungal species to the existing agrochemical fungicides has alarmed to the worldwide scientific community. There is no available methodology to predict in an efficient way if a new fungicide will have resistance risk due to fungal species which cause considerable crop losses. In an attempt to overcome this problem, a multi resistance risk QSAR model, based on substructural descriptors was developed from a heterogeneous database of compounds. The purpose of this model is the classification, design, and prediction of agrochemical fungicides according to resistance risk categories. The QSAR model classified correctly 85.11% of the fungicides and the 85.07% of the inactive compounds in the training series, for an accuracy of 85.08%. In the prediction series, the percentages of correct classification were 85.71 and 86.55% for fungicides and inactive compounds, respectively, with an accuracy of 86.39%. Some fragments were extracted and their quantitative contributions to the fungicidal activity were calculated taking into consideration the different resistance risk categories for agrochemical fungicides. In the same way, some fragments present in molecules with fungicidal activity and with negative contributions were analyzed like structural alerts responsible of resistance risk. PMID- 21633789 TI - Targeting tuberculosis through a small focused library of 1,2,3-triazoles. AB - Looking for new active molecules against Mycobacterium tuberculosis, a small focused library of 1,2,3-triazoles was efficiently prepared by click chemistry. Compounds were subsequently tested against different pathogenic and opportunistic mycobacteria including M. avium and M. tuberculosis. Two of them showed MIC at lower MUg/mL concentration for M. avium and even below that for M. tuberculosis, being more potent that control drugs. PMID- 21633790 TI - Ab initio DFT study of bisphosphonate derivatives as a drug for inhibition of cancer: NMR and NQR parameters. AB - DFT computations were carried out to characterize the (17)Oand (2)H electric field gradient, EFG, in various bisphosphonate derivatives. The computations were performed at the B3LYP level with 6-311++G (d,P) standard basis set. Calculated EFG tensors were used to determine the (17)O and (2)H nuclear quadrupole coupling constant, chi and asymmetry parameter, eta. For better understanding of the bonding and electronic structure of bisphosphonates, isotropic and anisotropic NMR chemical shieldings were calculated for the (13)C, (17)O and (31)P nuclei using GIAO method for the optimized structure of intermediate bisphosphonates at B3LYP level of theory using 6-311++G (d, p) basis set. The results showed that various substituents have a strong effect on the nuclear quadrupole resonance (NQR) parameters (chi, eta) of (17)O in contrast with (2)H NQR parameters. The NMR and NQR parameters were studied in order to find the correlation between electronic structure and the activity of the desired bisphosphonates. In addition, the effect of substitutions on the bisphosphonates polarity was investigated. Molecular polarity was determined via the DFT calculated dipole moment vectors and the results showed that substitution of bromine atom on the ring would increase the activity of bisphosphonates. PMID- 21633791 TI - Mucosal immunization provides better protection than subcutaneous immunization against Pasteurella multocida (B:2) in mice preimmunized with the outer membrane proteins. AB - To investigate the effect of boosting immunity via mucosal route vis-a-vis parenteral route in the mouse model of haemorrhagic septicaemia, mice preimmunized with OMP of Pasteurella multocida (B:2) were immunized with 10(2) cfu of P. multocida via intranasal and subcutaneous routes. Mice were challenged through intranasal route (natural route of infection) with 10(8) cfu 14 days after immunization. Group of mice which were immunized intranasally showed significant protection (P < 0.05) of 88% as compared to 50% protection in group of mice immunized subcutaneously. In the control group of mice, 100% mortality occurred within 48 h. of challenge. The results of present study indicated that boosting of immunity via mucosal route in mice preimmunized with OMP provided better protection against P. multocida. This study may have implications for developing better vaccination strategies for the natural host. PMID- 21633792 TI - Anterior and posterior vertebral column resection for severe and rigid idiopathic scoliosis. AB - A total of 16 patients with severe and rigid idiopathic scoliosis treated by anterior and posterior vertebral column resection (APVCR) were retrospectively reviewed after a minimum follow-up of 2 years. The indication for APVCR was scoliosis more than 90 degrees with flexibility less than 20%. The radiographic parameters were evaluated, and clinical records were reviewed. All patients underwent APVCR with posterior pedicle screw instrumentation in a two-stage surgery. The rib hump was reduced from 7.2 cm preoperatively to 1.8 cm at final follow-up (75% correction). Preoperative curves ranged from 93 degrees to 110 degrees Cobb angle. Coronal plane correction of the major curve averaged 67% with an average loss of correction of 1.4%. The apical vertebral translation of the major curve was corrected by 63.5%. The preoperative coronal imbalance of 0.9 cm (range 0-2.4) was improved to 0.8 cm (range 0.1-1.7) at the most recent follow up. The preoperative sagittal imbalance of 1.0 cm (range -3.1 to 4.6) was improved to 0.9 cm (range -2.6 to 3.0) at the most recent follow-up. Complications were encountered in four patients. One patient required ventilator support for 12 h after anterior surgery. Malposition of one pedicle screw was found in one patient. Malposition of titanium mesh cage happened to two patients. There were no neurological complications, deep wound infections or pseudarthrosis. APVCR is an effective alternative for severe and rigid idiopathic scoliosis. PMID- 21633793 TI - Etanercept attenuates pain-related behavior following compression of the dorsal root ganglion in the rat. AB - PURPOSE: TNFalpha is an inflammatory mediator related to neuropathic pain including sciatica. Much basic research suggests that anti-TNFalpha therapy may be useful for the treatment of sciatica. The purpose of this study was to clarify the effects of etanercept in a dorsal root ganglion (DRG) compression model. METHODS: Adult male Sprague-Dawley rats (200-250 g, n = 60) were used. An L shaped stainless rod was used to compress the left L5 DRG in the saline and etanercept groups. No rod was used in the sham group. In the etanercept group, 1 mg of etanercept was applied locally onto the DRG at the end of surgery. Saline was applied in the saline and sham groups. On day 3 and day 7 after surgery, the number of ED1-immunoreactive (IR) cells (macrophages) in the DRG was calculated by immunohistochemical methods (n = 6). In addition, double-immunofluorescence labeling for ED1 and TNFalpha was performed. Behavioral testing with von Frey filaments and a heat stimulator was performed (n = 12). RESULTS: ED1-IR cells in the DRG significantly increased in the control group compared with the sham group (p < 0.05). Some ED1-IR cells were co-labeled for TNFalpha. In the etanercept group, decrease in mechanical threshold was significantly inhibited compared with the saline group (p < 0.05). Thermal hyperalgesia was observed in the control group, but in neither the sham nor etanercept group (p < 0.05). CONCLUSION: Etanercept attenuated the pain-related behavior induced by DRG compression. These findings suggest that mechanical effects on the DRG might be reduced by etanercept in addition to the effects on nucleus pulposus in lumbar disc herniation. PMID- 21633795 TI - The assessment of spatial distribution of soil salinity risk using neural network. AB - Soil salinity in the Aral Sea Basin is one of the major limiting factors of sustainable crop production. Leaching of the salts before planting season is usually a prerequisite for crop establishment and predetermined water amounts are applied uniformly to fields often without discerning salinity levels. The use of predetermined water amounts for leaching perhaps partly emanate from the inability of conventional soil salinity surveys (based on collection of soil samples, laboratory analyses) to generate timely and high-resolution salinity maps. This paper has an objective to estimate the spatial distribution of soil salinity based on readily or cheaply obtainable environmental parameters (terrain indices, remote sensing data, distance to drains, and long-term groundwater observation data) using a neural network model. The farm-scale (~15 km(2)) results were used to upscale soil salinity to a district area (~300 km(2)). The use of environmental attributes and soil salinity relationships to upscale the spatial distribution of soil salinity from farm to district scale resulted in the estimation of essentially similar average soil salinity values (estimated 0.94 vs. 1.04 dS m(-1)). Visual comparison of the maps suggests that the estimated map had soil salinity that was uniform in distribution. The upscaling proved to be satisfactory; depending on critical salinity threshold values, around 70-90% of locations were correctly estimated. PMID- 21633796 TI - Adapting forest health assessments to changing perspectives on threats--a case example from Sweden. AB - A revised Swedish forest health assessment system is presented. The assessment system is composed of several interacting components which target information needs for strategic and operational decision making and accommodate a continuously expanding knowledge base. The main motivation for separating information for strategic and operational decision making is that major damage outbreaks are often scattered throughout the landscape. Generally, large-scale inventories (such as national forest inventories) cannot provide adequate information for mitigation measures. In addition to broad monitoring programs that provide time-series information on known damaging agents and their effects, there is also a need for local and regional inventories adapted to specific damage events. While information for decision making is the major focus of the health assessment system, the system also contributes to expanding the knowledge base of forest conditions. For example, the integrated monitoring programs provide a better understanding of ecological processes linked to forest health. The new health assessment system should be able to respond to the need for quick and reliable information and thus will be an important part of the future monitoring of Swedish forests. PMID- 21633797 TI - Occurrence and dissipation of veterinary antibiotics in two typical swine wastewater treatment systems in east China. AB - The occurrence and dissipation of 14 selected antibiotics comprising tetracyclines, sulfonamides, macrolides, fluoroquinolones, and chloramphenicols were investigated in two swine wastewater treatment systems in east China in three sampling surveys. The compounds were extracted from wastewater samples by solid-phase extraction and analyzed by ultra-performance liquid chromatography coupled with tandem mass spectrometry. The aqueous samples were unavoidably contaminated with antibiotics and the target antibiotics present in high contaminations were tetracycline, oxytetracycline, chlortetracycline, doxycycline, and sulfadiazine, with maximum concentrations of the individual contaminants reaching 41.6 * 10(3), 23.8 * 10(3), 13.7 * 10(3), 685.6 * 10(3), and 98.8 * 10(3) ng/l, respectively. The concentration ranges of these compounds were in the same order of magnitude as previously reported values in China and elsewhere. Biological activity can significantly degrade the contaminants but showed low efficiencies of dissipation of other analytes at lower levels of contamination. The removal efficiencies varied for different compounds and depended on their physiochemical properties and the treatment processes utilized at each waste treatment plant. Some target antibiotics were present in feeds obtained from swine farms at a range of average concentrations between 0.1 and 46.8 MUg/kg. However, the extremely high levels of the main contaminants found in these wastewaters cannot be ascribed solely to the pattern of consumption of prescription feeds. PMID- 21633798 TI - A new system for 3D ultrasound-guided placement of cerebral ventricle catheters. AB - PURPOSE: We present a new system for 3D ultrasound-guided placement of cerebral ventricle catheters. The system has been developed with the aim to provide accurate ultrasound-based guidance with only minimal changes to the current surgical technique and workflow. METHODS: The system consists of a pre-calibrated navigation adapter for the catheter and a reference frame attached to a standard surgical retractor in addition to an ultrasound-based navigation system with a probe that fits on top of a standard burr hole. RESULTS: The accuracy of the pre calibrated system has been evaluated, and our measurements indicate that the accuracy of the pre-calibrated system is better than 3 mm. We also present a clinical case. CONCLUSIONS: The navigation accuracy is considered sufficient for clinical use, and initial clinical tests are promising. Further testing will be necessary to fully evaluate the performance of the system in a clinical setting. PMID- 21633799 TI - Comparing two approaches to rigid registration of three-dimensional ultrasound and magnetic resonance images for neurosurgery. AB - PURPOSE: We present a new technique for registering magnetic resonance (MR) and ultrasound images in the context of neurosurgery. It involves generating a pseudo ultrasound (pseudo-US) from a segmented MR image and uses cross-correlation as the cost function to register the pseudo-US to the real ultrasound data. The algorithm's performance is compared with that of a state-of-the-art technique that uses a median-filtered MR image to register to a Gaussian-blurred ultrasound using a normalized mutual information (NMI) objective function. METHODS: The two methods were tested on data from 15 patients with brain tumor, including low-and high-grade gliomas, in both first operations and reoperations. Two metrics were used to evaluate registration accuracy: (1) the mean distance between corresponding points, identified on both MR and ultrasound images by two experts, and (2) ratings based on visual comparison by one neurosurgeon. RESULTS: The mean residual distance of the pseudo-US technique, 2.97 mm, is significantly more accurate (p = .0011) than that of the NMI approach, 4.86 mm. The visual assessment shows that only 4 of the 15 cases had a satisfactory initial alignment based on homologous skin-point registration. There is a significant correlation between the quantitative distance measures and the qualitative ratings (rho = 0.785). CONCLUSION: The results show that the pseudo-US rigid registration technique robustly improves the MRI-ultrasound alignment when compared with the initial alignment, even when applied to highly distorted brains and a large range of tumor sizes and appearances. PMID- 21633800 TI - Utilisation of advance motor information is impaired in Friedreich ataxia. AB - Friedreich ataxia (FRDA) is the most common of the genetically inherited ataxias. We sought to examine motor planning ability in 13 individuals with FRDA and 13 age- and sex-matched control participants using two experimental paradigms that examined the ability to incorporate different levels of advance information to plan sequential movements. Individuals with FRDA demonstrated a differential pattern of motor response to advance information and were significantly disadvantaged by conditions requiring initiation of movement without a direct visual cue. There was also a significant negative correlation with age of disease onset and differing levels of advance information, suggesting an impact of FRDA on the development of motor cognition, independent of the effect of disease duration. We suggest that deficits are due to cerebellar impairment disrupting cerebro-ponto-cerebello-thalamo-cerebral loops (and thus cortical function), direct primary cortical pathology or a possible combination of the two. PMID- 21633801 TI - Linking animal movement to site fidelity. AB - Site fidelity, the recurrent visit of an animal to a previously occupied area is a wide-spread behavior in the animal kingdom. The relevance of site fidelity to territoriality, successful breeding, social associations, optimal foraging and other ecological processes, demands accurate quantification. Here we generalize previous theory that connects site fidelity patterns to random walk parameters within the framework of the space-time fractional diffusion equation. In particular, we describe the site fidelity function in terms of animal movement characteristics via the Levy exponent, which controls the step-length distribution of the random steps at each turning point, and the waiting time exponent that controls for how long an animal awaits before actually moving. The analytical results obtained will provide a rigorous benchmark for empirically driven studies of animal site fidelity. PMID- 21633802 TI - Inflammation and mortality in a frail mouse model. AB - Mice homozygous for targeted deletion of the interleukin 10 gene (Il-10) have been partially characterized as a model for human frailty. These mice have increased serum interleukin (IL)-6 in midlife, skeletal muscle weakness, and an altered skeletal muscle gene expression profile compared to age and sex-matched C57BL/6 (B6) control mice. In order to further characterize for use as a frailty model, we evaluated the evolution of inflammatory pathway activation, endocrine change, and mortality in these mice. Serum was collected in groups of age- and sex-matched B6.129P2-Il10(tm1Cgn)/J (IL-10(tm/tm)) mice and B6 control mice at age 12, 24, 48, 72, and 90 weeks. Cytokines including IL-6, interleukin 1 beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN gamma), chemokine (C-X-C motif) ligand 1 (KC), IL-12, and IL-10 were measured using electro-chemiluminescent multiplex immunoassay and insulin-like growth factor 1 (IGF-1) was measured using solid-phase enzyme-linked immunosorbent assay. A separate longitudinal cohort was monitored from age 35 weeks to approximately 100 weeks. Survival was evaluated by Kaplan-Meier survival estimates and detailed necropsy information was gathered in a subset of mice that died or were sacrificed. In IL-10(tm/tm) mice compared to B6 controls, serum IL 6, IL-1beta, TNF-alpha, IFN-gamma, KC levels were significantly elevated across the age groups, serum mean IGF-1 levels were higher in the 48-week-old groups, and overall mortality rate was significantly higher. The quadratic relationship between IGF-1 and age was significantly different between the two strains of mice. Serum IL-6 was positively associated with IGF-1 but the effect was significantly larger in IL-10(tm/tm) mice. These findings provide additional rationale for the use of the IL-10(tm/tm) mouse as a model for frailty and for low-grade inflammatory pathway activation. PMID- 21633803 TI - [Emergencies in gastroenterology]. PMID- 21633804 TI - [Acute Wilson disease]. AB - Wilson disease is an autosomal recessive inherited disorder of human copper metabolism clinically associated with hepatic damage and/or neurological symptoms of varying degree. Copper accumulation and toxicity result in direct injury to hepatocytes followed by inflammation and irreversible impairment of neurons, mainly in the extrapyramidal system. A not insignificant number of cases begin with fulminant liver failure or acute appearance of neurological symptoms. If left untreated or in the case of delayed diagnosis and treatment, both acute manifestations may result in irreversible symptoms or even death. Rapid and exact diagnosis by means of clinical, biochemical and genetic analysis and the immediate initiation of drug therapy with copper chelators or, in the case of fulminant liver failure, orthotopic liver transplantation are essential for a favourable outcome in patients with acute Wilson disease. PMID- 21633805 TI - Neuregulin in heart failure : reverse translation from cancer cardiotoxicity to new heart failure therapy. AB - Trastuzumab is a monoclonal antibody to the ErbB2 (Her2nue) receptor over expressed in Her2(+) breast cancer. Trastuzumab-related cardiotoxicity has revealed the importance of ErbB2 signaling in the heart. Neuregulin (NRG-1) is an important stress-mediated paracrine growth factor that signals through the family of ErbB receptors to promote cardioprotection (myocyte cell survival, proliferation, differentiation, hypertrophy, and angiogenesis). Animal models with disrupted NRG/ErbB signaling fail to develop normally or result in impaired cardiac function post-natally. Pre-clinical animal studies and early-phase human studies suggest that recombinant NRG-1 holds promise as a new therapy for the treatment of various forms of heart failure. Much work is needed to further understand the exact mechanisms of cardiac repair and to find a safe mode of application for recombinant NRG-1 in heart failure. PMID- 21633806 TI - Cardiac side effects of conventional and particle radiotherapy in cancer patients. AB - Radiation therapy, which has become an integral part of modern treatment strategies for many types of cancer in recent decades, is associated with the risk of long-term adverse effects. Particularly in the case of patients with a good prognosis as a result of these modern treatment modalities, avoiding therapy related side effects plays an increasingly important role. Of these side effects, cardiac complications are particularly relevant since they not only adversely affect quality of life but can also be potentially life-threatening. This review summarizes and critically analyzes the available evidence on the impact of ionizing irradiation on the cardiovascular system. The paper presents an overview of the current knowledge on epidemiological evidence, clinical presentation and risk factors of radiation-induced cardiovascular disease. Additionally, mechanisms of development of this type of complication and strategies for screening, treatment, and avoiding cardiac complications are discussed. PMID- 21633807 TI - Comparing deep anterior lamellar keratoplasty and automated lamellar therapeutic keratoplasty in patients with keratoconus. AB - PURPOSE: We compared outcomes of deep anterior lamellar keratoplasty (DALK) and automated lamellar therapeutic keratoplasty (ALTK) in patients with keratoconus. METHODS: Seventy eyes with keratoconus that underwent corneal transplantation using either DALK (n = 40) or ALTK (n = 30) were included in this retrospective study. DALK was performed with Anwar's big-bubble technique and ALTK using an ALTK system. Best-corrected visual acuity (BCVA), refractive results, and complications were analyzed. RESULTS: Mean follow-up in the DALK (16.68 +/- 3.36 months) and ALTK (15.27 +/- 2.50 months) groups were similar (p = 0.058). Mean final BCVA of patients in the DALK group was 0.31 +/- 0.14 logarithm of the minimum angle of resolution (logMAR) and in the ALTK group 0.34 +/- 0.15 logMAR, (p = 0.492). Corresponding figures for final postoperative spherical equivalents were -3.45 +/- 0.77 D and -4.19 +/- 0.99 D, respectively (p = 0.001). Final postoperative degree of astigmatism was similar in the two groups: DALK -4.28 +/- 0.66 D; ALTK -4.50 +/- 1.05 D (p = 0.307). CONCLUSION: Final visual acuity outcomes were comparable for the DALK and ALTK groups. Thus, ALTK seem to be as efficacious as DALK for surgical treatment of keratoconus. PMID- 21633808 TI - Intravitreal bevacizumab injection for peripheral exudative hemorrhagic chorioretinopathy. PMID- 21633809 TI - A case of retinal light damage by green laser pointer (Class 3b). PMID- 21633810 TI - Efficacy and safety of single-dose 2.0 g azithromycin in the treatment of acute exacerbation of chronic obstructive pulmonary disease. AB - To evaluate the efficacy and safety of single-dose 2.0 g azithromycin (ZSR) in the treatment of acute exacerbation of chronic obstructive pulmonary disease (AE COPD), we retrospectively reviewed all patients with AE-COPD who were treated with ZSR. In comparison with patients who received intravenous therapy for AE COPD, the clinical cure rate, length of stay in hospital, and medical costs were evaluated. A total of 29 patients thus were eligible for this study. Clinical cure rates of ZSR and intravenous therapy for the treatment of AE-COPD were 83.3% (n = 12) and 88.2% (n = 17), respectively, between the groups (P = 1.000). No severe adverse events were found in either group. The ZSR and intravenous groups averaged 9.9 and 12.5 days of admission, respectively. Length of admission for clinical success cases was much shorter for patients who received ZSR than patients who received intravenous therapy (6.2 vs. 11.9 days, P = 0.038). Medical costs were less for the group receiving ZSR than for the intravenous therapy group. We suggest ZSR can achieve near-perfect compliance and could be one of the tools in the treatment of AE-COPD. PMID- 21633811 TI - The polyphenol oxidase gene family in poplar: phylogeny, differential expression and identification of a novel, vacuolar isoform. AB - Polyphenol oxidases (PPOs) are oxidative enzymes that convert monophenols and o diphenols to o-quinones using molecular oxygen. The quinone products are highly reactive following tissue damage and can interact with cellular constituents and cause oxidative browning and cross-linking. The induction of PPO in some plants as a result of wounding, herbivore attack, or pathogen infection has implicated them in defense. However, PPO-like enzymes that act as specific hydroxylases, for example in lignan and pigment biosynthesis, have also been discovered. Here, we present the first genome-enabled analysis of a PPO gene family. The Populus trichocarpa genome was found to contain a minimum of nine complete PPO genes, and seven of these were characterized further. The PPO gene family includes both recently duplicated and divergent sequences that are 36-98% identical at the amino acid level. Gene expression profiling in poplar tissues and organs revealed that the PPO genes are all differentially expressed during normal development, but that only a small subset of PPO genes are significantly upregulated by wounding, methyl jasmonate or pathogen infection. Our studies also identified PtrPPO13, a novel PPO gene that is predicted to encode an N-terminal signal peptide. Transient expression of green fluorescent protein fusions demonstrated its localization to the vacuolar lumen. Together, our findings show that the poplar PPO family is diverse and is likely linked to diverse physiological functions. PMID- 21633813 TI - Successful allogeneic hematopoietic stem cell transplantation for acute myeloid leukemia during respiratory failure and invasive mechanical ventilation. AB - Allogeneic hematopoietic stem cell transplantation (HSCT) is a curative treatment option for various hematologic disorders. However, life-threatening adverse events resulting from treatment-related toxicity, severe infections, and/or graft versus-host disease (GvHD) can occur. We report on a 64-year-old patient suffering from secondary acute myeloid leukemia (AML) who underwent successful allogeneic HSCT while on invasive mandatory ventilation (IMV). The patient received reduced intensity conditioning (RIC) according to the FLAMSA-protocol. Acute respiratory failure occurred one day before scheduled HSCT. Following emergency endotracheal intubation the patient was transferred to the intensive care unit (ICU). Because of respiratory deterioration, stem cell infusion was postponed. After stabilization of respiratory parameters, HSCT was performed during IMV which was continued for seven days. Following hematopoietic regeneration the patient was discharged in good condition on day 35 after HSCT. This case illustrates that intubation and mechanical ventilation do not necessarily exclude leukemic patients from HSCT. PMID- 21633814 TI - Endoparasites of cats from the Tirana area and the first report on Aelurostrongylus abstrusus (Railliet, 1898) in Albania. AB - Following the recovery of first-stage nematode larvae indicative of Aelurostrongylus abstrusus infection in the faeces of free-roaming cats from the greater Tirana area, examination of 18 cats at necropsy revealed nine of them harbouring adult A. abstrusus (Railliet, 1898) in the lungs (prevalence, 50%; range, 1-11). In addition to A. abstrusus, Eucoleus aerophilus (16.7%; 1-9) was isolated from the lungs, and Toxocara cati (83.3%; 2-33), Ancylostoma tubaeforme (44.4%; 1-20), Dipylidium caninum (83.3%; 1-164), Joyeuxiella pasqualei (11.1%; 1 3) and one specimen of an acanthocephalan (5.5%) were recovered from the gastrointestinal tract. Furthermore, oocysts of Cystoisospora felis and C. rivolta were found in the rectal faeces of 5.6% and 11.1% of the cats, respectively. In conclusion, the prevalence of endoparasite infection in free roaming cats in Tirana can be considered to be high. The occurrence of A. abstrusus, which may cause respiratory distress in cats, is reported for the first time in Albania. PMID- 21633816 TI - Metallothionein protein evolution: a miniassay. AB - Metallothionein (MT) evolution is one of the most obscure yet fascinating aspects of the study of these atypical metal-binding peptides. The different members of the extremely heterogeneous MT protein superfamily probably evolved through a web of duplication, functional differentiation, and/or convergence events leading to the current scenario, which is particularly hard to interpret in terms of molecular evolution. Difficulties in drawing straight evolutionary relationships are reflected in the lack of definite MT classification criteria. Presently, MTs are categorized either according to a pure taxonomic clustering or depending on their metal binding preferences and specificities. Extremely well documented MT revisions were recently published. But beyond classic approaches, this review of MT protein evolution will bring together new aspects that have seldom been discussed before. Hence, the emergence of life on our planet, since metal ion utilization is accepted to be at the root of the emergence of living organisms, and global trends that underlie structural and functional MT diversification, will be presented. Major efforts are currently being devoted to identifying rules for function-constrained MT evolution that may be applied to different groups of organisms. PMID- 21633817 TI - Informal caring-time and caregiver satisfaction. AB - This paper examines the role of care decision processes on informal caring-time choices. We focus on three care decisions: the caregiver's own decision, a family decision and a recipient request. Results show that informal caregivers, engaged in care activities as a result of a family decision, are more likely to devote more than 5 h to care activities, even after allowing for endogeneity. Our findings are robust to controlling for a large number of socio-demographic characteristics, including care recipient and caregiver characteristics. Supplemental analysis, developed to explore whether care arrangements are related to informal caregiver's satisfaction, indicates that the family decision heavily penalizes informal caregivers. Given the importance of informal care activities in reducing health care costs, our findings imply that care decision processes should be taken into consideration when formulating health care policies. PMID- 21633818 TI - A proposed model for economic evaluations of major depressive disorder. AB - In countries like UK and Australia, the comparability of model-based analyses is an essential aspect of reimbursement decisions for new pharmaceuticals, medical services and technologies. Within disease areas, the use of models with alternative structures, type of modelling techniques and/or data sources for common parameters reduces the comparability of evaluations of alternative technologies for the same condition. The aim of this paper is to propose a decision analytic model to evaluate long-term costs and benefits of alternative management options in patients with depression. The structure of the proposed model is based on the natural history of depression and includes clinical events that are important from both clinical and economic perspectives. Considering its greater flexibility with respect to handling time, discrete event simulation (DES) is an appropriate simulation platform for modelling studies of depression. We argue that the proposed model can be used as a reference model in model-based studies of depression improving the quality and comparability of studies. PMID- 21633819 TI - Different effects of intermittent and continuous fluid shear stresses on osteogenic differentiation of human mesenchymal stem cells. AB - A reasonable mechanical microenvironment similar to the bone microenvironment in vivo is critical to the formation of engineering bone tissues. As fluid shear stress (FSS) produced by perfusion culture system can lead to the osteogenic differentiation of human mesenchymal stem cells (hMSCs), it is widely used in studies of bone tissue engineering. However, effects of FSS on the differentiation of hMSCs largely depend on the FSS application manner. It is interesting how different FSS application manners influence the differentiation of hMSCs. In this study, we examined the effects of intermittent FSS and continuous FSS on the osteogenic differentiation of hMSCs. The phosphorylation level of ERK1/2 and FAK is measured to investigate the effects of different FSS application manners on the activation of signaling molecules. The results showed that intermittent FSS could promote the osteogenic differentiation of hMSCs. The expression level of osteogenic genes and the alkaline phosphatase (ALP) activity in cells under intermittent FSS application were significantly higher than those in cells under continuous FSS application. Moreover, intermittent FSS up regulated the activity of ERK1/2 and FAK. Our study demonstrated that intermittent FSS is more effective to induce the osteogenic differentiation of hMSCs than continuous FSS. PMID- 21633820 TI - Bacteriocin release protein-mediated secretory expression of recombinant chalcone synthase in Escherichia coli. AB - Flavonoids are secondary metabolites synthesized by plants shown to exhibit health benefits such as anti-inflammatory, antioxidant, and anti-tumor effects. Thus, due to the importance of this compound, several enzymes involved in the flavonoid pathway have been cloned and characterized in Escherichia coli. However, the formation of inclusion bodies has become a major disadvantage of this approach. As an alternative, chalcone synthase from Physcomitrella patens was secreted into the medium using a bacteriocin release protein expression vector. Secretion of P. patens chalcone synthase into the culture media was achieved by co-expression with a psW1 plasmid encoding bacteriocin release protein in E. coli Tuner (DE3) plysS. The optimized conditions, which include the incubation of cells for 20 h with 40 ng/ml mitomycin C at OD(600) induction time of 0.5 was found to be the best condition for chalcone synthase secretion. PMID- 21633821 TI - Selenium is significantly depleted among morbidly obese female patients seeking bariatric surgery. AB - BACKGROUND: The aim of this study was to compare serum levels of trace elements in morbidly obese female patients seeking bariatric surgery with those of age matched females with body mass index (BMI) less than or equal to 30. METHODS: Blood samples were collected from 66 morbidly obese female patients seeking bariatric surgery prior to undergoing surgery. Blood was collected also from 44 female patients (with BMI less than or equal to 30) prospectively from April 2009 till February 2010. Exclusion criteria in both groups were the presence of any co morbidities on medication, patients receiving any vitamin supplement or any herbal product intake, smoking, and alcohol consumption. Serum zinc, magnesium, copper, and selenium were measured and compared between the two groups. RESULTS: The mean age and BMI for morbidly obese female patients was 28.5 years (21.5 35.5) and 45.26 kg/m(2) (40.3-50.22). The control group had a mean age and BMI of 30.75 years (21.35-40.15) and 25.88 kg/m(2) (22.73-29.03). For morbidly obese patients, the serum level of copper, zinc, selenium, and magnesium was 1,623.84, 698.34, 86.08, and 17,830 MUg/l, respectively, compared to 1,633.36, 734.82, 101.14, and 18,260 MUg/l for the control group. The serum levels of the trace elements were not statistically significantly different between the two groups except for selenium, which was significantly reduced among morbidly obese female patients (p < 0.0001). CONCLUSIONS: Serum selenium level is significantly reduced among morbidly obese female patients seeking bariatric surgery. PMID- 21633822 TI - Possible consequences of the new operation criteria for bariatric surgery in Switzerland. PMID- 21633823 TI - Levothyroxine absorption in morbidly obese patients before and after Roux-En-Y gastric bypass (RYGB) surgery. AB - BACKGROUND: Roux-en-Y gastric bypass (RYGB) modifies the anatomical structure of the upper intestine tract, reduces gastric acid secretion, and may impair LT4 absorption. The aim of this study was to evaluate the LT4 absorption in morbidly obese patients before and after RYGB. METHODS: Thirty morbidly obese patients were divided in two groups: The NS group included 15 patients before RYGB surgery (BMI = 43.1 +/- 4 kg/m(2)), and the S group included 15 patients after surgery (BMI = 37.3 +/- 4 kg/m(2)). Two baseline samples were collected, and 600 MUg of oral LT4 tablets were administered. Blood samples were collected at 30, 60, 120, 180, 240, 300, and 1440 min. Serum-free T4 (FT4), total T4 (TT4), and TSH were measured at each time point. The increase in TT4, FT4, and TSH (DeltaTT4, DeltaFT4, and DeltaTSH) was calculated, subtracting from the baseline mean value. RESULTS: The pharmacokinetics parameters regarding LT4 absorption, maximum DeltaTT4, and area under the curve(AUC) of both DeltaTT4 and DeltaFT4 were significantly higher in the S group compared with the NS group (p < 0.05). It was observed, however, that there was a significant delay in the absorption of LT4 in the S group. Basal serum TSH and leptin levels were higher in the NS group (p = 0.016 and 0.026, respectively), whereas basal serum TT4, FT4, DeltaTSH, and the AUC of DeltaTSH were similar between groups. CONCLUSIONS: In this study, we have demonstrated that Roux-en-Y bypass surgery does not diminish LT4 absorption. A small but significant delayed absorption of LT4, however, was observed in patients after surgery. PMID- 21633824 TI - Clinical significance of vagus nerve variation in radiofrequency ablation of thyroid nodules. AB - OBJECTIVES: To evaluate the types and incidence of vagus nerve variations and to assess factors related to the vulnerability of vagus nerves during the radiofrequency (RF) ablation of thyroid nodules. METHODS: Bilateral vagus nerves of 304 consecutive patients who underwent ultrasound of the neck were assessed. Two radiologists evaluated vagus nerve type (types 1-4; lateral/anterior/medial/posterior), the shortest distance between the thyroid gland and vagus nerve, and thyroid contour. Vagus nerve vulnerability was defined as a vagus nerve located within 2 mm of the thyroid gland through the ex vivo experiments, and factors associated with vulnerability were assessed. RESULTS: We were unable to find one vagus nerve. Of the 607 vagus nerves, 467 (76.9%) were type 1, 128 (21.1%) were type 2, 10 (1.6%) were type 3, and 2 (0.3%) were type 4, with 81 (13.3%) being vulnerable. Univariate analysis showed that sex, location, thyroid contour and type were significantly associated with vagus nerve vulnerability. Multivariate analysis showed that bulging contour caused by thyroid nodules (P = 0.001), vagus nerve types 2/4 (P < 0.001) and type 3 (P < 0.001) were independent predictors. CONCLUSION: The operator should pay attention to anatomical variations and the resulting vagus nerve injury during RF ablation of bulging thyroid nodules. PMID- 21633825 TI - Radiation dose threshold for coronary artery calcium score with MDCT: how low can you go? AB - OBJECTIVES: To evaluate the lowest radiation exposure threshold at which coronary calcium scoring (CCS) remains accurate. METHODS: A prospective study of 43 consecutive eligible patients referred for CCS underwent imaging in accordance with the manufacturer's recommended protocol. Dedicated software was used to generate 8 series of images simulating tube currents ranging from 20 to 300 mA. These images were randomised and read in blinded fashion to determine the lowest tube current at which the CCS remained accurate. The minimum mA was correlated with 6 different patients' biometric parameters: bodyweight, body mass index, AP and lateral thoracic diameters, average thoracic diameter and the scout attenuation coefficient (SAC). The 95% confidence interval for each parameter was used to calculate tube current threshold levels and hence stratified CCS protocols were derived. RESULTS: Spearman's correlation coefficients of the minimum tube current for the 6 parameters were: 0.66, 0.63, 0.65, 0.74, 0.77 and 0.86 respectively (p < 0.001). SAC offered the largest potential reduction in mean effective dose from 1.86 mSv to 0.88 mSv. CONCLUSION: CCS with at least 50% reduction in radiation exposure and below 1 mSv is feasible if CT scout projections are utilised effectively. PMID- 21633826 TI - Role of contrast enhanced ultrasound in acute scrotal diseases. AB - OBJECTIVE: To evaluate the efficacy of contrast-enhanced ultrasound (CEUS) in patients with acute scrotal pain not defined at ultrasound (US) with colour Doppler . METHODS: CEUS was carried out in 50 patients with acute scrotal pain or scrotal trauma showing testicular lesion of undefined nature at US. The accuracy of US and CEUS findings versus definitive diagnosis (surgery or follow-up) was calculated. RESULTS: Twenty-three patients had a final diagnosis of testicular tumour, three abscess, eight focal infarction, seven trauma, three testicular torsion, one haematoma. Five patients were negative. Thirty-five patients were operated (23 testicular tumours, six trauma, three testicular torsion, one abscess, one focal infarction, and one haematoma) and 15 underwent medical treatment or were discharged. US provided a definitive diagnosis in 34/50 as compared to the 48/50 patients diagnosed at CEUS. Sensitivity and specificity were 76% and 45% for US and 96% and 100% for CEUS respectively. CONCLUSIONS: CEUS was more accurate in the final diagnosis compared to US, potentially reducing the need for further imaging. In particular CEUS can be proposed in emergency in cases where US diagnosis remains inconclusive, namely in infarction, and trauma, when testicular torsion cannot be ruled out, and in identifying testicular mass. PMID- 21633827 TI - The calcium scare--what would Austin Bradford Hill have thought? AB - Detailed consideration of the suggested association between calcium supplementation and heart attacks has revealed weakness in the evidence which make the hypothesis highly implausible. INTRODUCTION: The aim of this study was to evaluate the strength of the evidence that calcium supplementation increases the risk of myocardial infarction. METHODS: This study used critical examination of a meta-analysis of the effects of calcium supplements on heart attacks in five prospective trials on 8,016 men and women, and consideration of related publications by the same author. RESULTS: The meta-analysis was found to be subject to several limitations including non-adherence to the clinical protocol, multiple endpoint testing and failure to correctly adjust for endpoint ascertainment. The main risk factors for myocardial infarction were not available for 65% of the participants, and none of the trials had cardiovascular disease as its primary endpoint. There were more overweight participants, more subjects on thyroxine and more men on calcium than on placebo. In particular, over 65% of all the heart attacks were self-reported. When the evidence was considered in the light of Austin Bradford Hill's six main criteria for disease causation, it was found not to be biologically plausible or strong or to reflect a dose-response relationship or to be consistent or to reflect the relationship between the trends in calcium supplementation and heart attacks in the community or to have been confirmed by experiment. The addition of a more recent trial on 1,460 women over 5 years reduced the relative risk to 1.23 (P = 0.0695). CONCLUSION: Present evidence that calcium supplementation increases heart attacks is too weak to justify a change in prescribing habits. PMID- 21633828 TI - Diagnostic capabilities of fractal dimension and mandibular cortical width to identify men and women with decreased bone mineral density. AB - Dental panoramic radiographs could be used to screen for osteopenia. We found the fractal dimension to be a good discriminator of osteopenia in both men and women but that the mandibular cortical width (MCW) did not perform as well in men. The fractal dimension may be a valid screening tool. INTRODUCTION: The aim of this study was to assess the diagnostic capability of the fractal dimension and MCW measured from dental panoramic radiographs in identifying men and women with decreased bone mineral density (BMD). METHODS: The MCW and fractal dimension were measured from dental panoramic radiographs as surrogates for BMD. These measures were then compared to the results from dual-energy X-ray absorptiometry (DXA) performed for clinical purposes. A total of 56 subjects with the panoramic radiograph taken within 6 months of the DXA exam were used in the analysis for this study. RESULTS: The area under the curve of the fractal dimension for identifying low BMD (T-score <-1.0) was 0.81 (0.67, 0.95) and 0.78 (0.49, 1.00) for men and women, respectively. For the MCW, the area under the curve was found to be 0.53 (0.34, 0.72) and 0.80 (0.58, 1.00) for men and women, respectively. CONCLUSIONS: In this largely male study population, the fractal dimension was found to be a good discriminator of low BMD in both men and women. The MCW did not perform as well in men. PMID- 21633829 TI - [Basal cell carcinomas in radiotherapy-treated port wine stains. An ideal indication for Mohs surgery]. AB - BACKGROUND: Early treatment of port wine stains with ionizing radiation can lead to the development of often multifocal basal cell carcinomas (BCC) after decades. In most cases it is clinically impossible to distinguish between the tumor and the underlying vascular malformation and to decide where to set surgical margins. PATIENTS AND METHODS: We report on a series of three patients with BCC overlying a port wine stain that had previously been treated with radiation therapy in early childhood. In all patients Mohs surgery was performed to insure complete excision of the BCC. RESULTS: In our patients, development of BCC occurred about 20 to 40 years after radiation therapy. Clinically - without the help of Mohs surgery - the borders of the BCC could not have been detected due to the underlying nevi flammei in all cases. CONCLUSIONS: BCC overlying a port wine stain previously treated with radiation therapy is a rare but ideal indication for Mohs surgery. In addition we recommend regular clinical follow-ups to detect recurrent or additional BCC as early as possible. PMID- 21633830 TI - [Use of joint crisis plans in psychiatric hospitals in Germany: results of a nationwide survey]. AB - OBJECTIVE: Joint crisis plans in psychiatry are consensus-oriented instruments for patients to state their will in case of a crisis. The aim of this survey is to determine the prevalence, practice and barriers for implementation of joint crisis plans in Germany. METHODS: In spring 2009, 366 psychiatric hospitals in Germany were asked to complete a questionnaire. RESULTS: The return rate was 46.4%; 68% of the participating hospitals offered joint crisis plans. The number of completed joint crisis plans was low (median 2.7/year per clinic). The demand did not increase according to a majority of the hospitals. The main reason for hospitals to abstain from joint crisis plans was low demand. Hospitals providing the option of joint crisis plans reported positive experiences. CONCLUSIONS: A theoretical offer of joint crisis plans is widespread in German psychiatric hospitals. Despite positive experiences, patients do not ask for them frequently. PMID- 21633831 TI - High prevalence of upper urinary tract involvement detected by 111indium-oxine leukocyte scintigraphy in patients with candiduria. AB - The purpose of this investigation was to assess the prevalence of upper urinary tract involvement in patients with candiduria by means of (111)indium-oxine labeled leukocyte scintigraphy. An observational cohort study of patients with confirmed candiduria was conducted in an acute-care teaching hospital in Spain from March 2006 through February 2009. An (111)In-labeled leukocyte scan was performed in order to assess the upper urinary tract involvement. A series of non matched patients without candiduria nor bacteriuria undergoing scintigraphy to exclude infections in other sites than the urinary tract was also studied. Demographics, baseline illness, and clinical data were recorded. Candiduria was detected in 428 patients, and scintigraphy was performed in 35 of these patients. Twenty-nine patients without candiduria nor bacteriuria were also studied. Positive renal scintigraphy was documented in 24 (68%) patients with confirmed candiduria and in 3 (10%) patients without candiduria (p < 0.005). Renal uptake was not associated with a higher mortality nor with re-admissions. Subclinical pyelonephritis could be more frequent in patients with candiduria than it has been previously considered. PMID- 21633832 TI - Serum sTREM-1 as a surrogate marker of treatment outcome in patients with peptic ulcer disease. AB - OBJECTIVE: Soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) is elevated in the gastric juice and in cultures of gastric mucosa of patients with peptic ulcer disease (PUD). Its application as a surrogate marker for the treatment of PUD was assessed. METHODS: From 138 eligible patients, 96 were enrolled; 50 with duodenal ulcer, 29 with gastric ulcer and 17 with chronic gastritis. Patients were endoscoped twice; once before treatment and once after treatment. Biopsy specimens were collected for histopathologic estimation of gastritis. Blood was sampled prior to each endoscopy. Serum was collected and sTREM-1 was measured by an enzyme immunoabsorbent assay ( http://www.clinicaltrials.gov identifier NCT00534443). RESULTS: At the end of treatment sTREM-1 was either: (a) below the limit of detection (this occurred in 62 patients and it was accompanied by lacks signs of residual disease in 58 patients, 93.5%); or (b) above the limit of detection (this occurred in 17 patients and it was accompanied by residual disease in 14 patients, 82.3%) (p < 0.0001). Odds ratio for complete healing of peptic ulcer with sTREM-1 below detection limit was 5.30 (95% CI: 1.89-14.83, p < 0.001) compared to serum sTREM 1 above the limit of detection. CONCLUSIONS: Serum sTREM-1 below detection limit may effectively distinguish patients who successfully completed therapy for PUD from those with residual disease and apply as a surrogate marker. PMID- 21633833 TI - Retreatment of hepatitis C with consensus interferon and ribavirin after nonresponse or relapse to pegylated interferon and ribavirin: a national VA clinical practice study. AB - BACKGROUND: Studies of the retreatment with consensus interferon (CIFN) and ribavirin (RBV) of hepatitis C virus (HCV)-infected patients who failed prior pegylated interferon alfa/ribavirin (PEG-IFN/RBV) have found quite variable efficacy and tolerability of this therapy. As such, CIFN/RBV use and efficacy in clinical practice were evaluated within the Department of Veterans Affairs (VA), the largest national, integrated system for HCV care. AIMS: The purpose of this study was to determine rates of sustained virologic response (SVR) and patterns of CIFN/RBV use in the VA. Methods included retrospective review of national VA data in HCV-infected patients who had previously failed>=12 weeks of PEG-IFN/RBV and were prescribed CIFN/RBV between October 1, 2003 and September 30, 2006. RESULTS: A total of 597 patients met the study criteria. CIFN was primarily dosed as 15 mcg subcutaneously daily combined with standard doses of RBV. Mean treatment duration was 21 weeks; CIFN was discontinued within 4 weeks in 24%. Hematological growth factors were used in 49%. Post-treatment viral loads were available in 385 patients. SVR to CIFN/RBV was achieved in 11%, and was significantly higher in prior PEG-IFN/RBV relapsers compared with nonresponders (31% vs. 6%, respectively; P<0.0001). A 2-log10 or greater drop in HCV RNA after 24 weeks of PEG-IFN/RBV was a predictor of subsequent SVR to CIFN/RBV. CONCLUSIONS: CIFN/RBV was used frequently in clinical practice for retreatment of PEG-IFN/RBV. In this setting, early treatment discontinuation was common. Overall SVR was low, although response was significantly better in prior PEG-IFN/RBV relapsers and those who had a 2-log(10) or greater decline than in nonresponders. PMID- 21633834 TI - Neonatal outcomes and mental illness, substance abuse, and intentional injury during pregnancy. AB - Mental illness (MI), substance abuse (SA), and intentional injury (II) are known individual risk factors for adverse pregnancy outcomes. Their combined association with preterm birth (PTB) and low birth weight (LBW) remains relatively unexplored. We examined hospital utilization for the co-occurrence of II and MI or SA in pregnant women in Massachusetts and assessed their interactive association with PTB and LBW. This retrospective cohort study used ICD-9 and E codes reported on linked birth and hospital utilization data to identify MI, SA, and II diagnoses during pregnancy for 176,845 Massachusetts resident women who delivered during 2002-2004. Adjusted odds ratios (OR) for the independent and joint associations of MI, SA, and II on PTB and LBW were calculated. Two thousand two hundred and eight women (1.6%) had a prenatal MI visit, 834 (0.5%) a prenatal SA visit, and 847 (0.5%) a prenatal II visit. Among them 163 women had MI and II visits and 69 had SA and II visits. SA, MI, and II were all significant predictors of LBW and PTB. Women with both SA and II had higher odds of PTB (OR 2.7 95% CI 1.3-5.7) and LBW (OR 5.3 95% CI 3.9-7.3) than women with neither diagnosis. Prenatal MI, SA, and II are risk factors for LBW and PTB. Women with SA and II co-diagnoses have greater risk of LBW and PTB than women with neither diagnosis. Screening, timely diagnosis, and treatment of women with co-occurring morbidities, particularly II and SA, should be incorporated into reproductive and perinatal health programs. PMID- 21633835 TI - Effect of selenium on connexin expression, angiogenesis, and antioxidant status in diabetic wound healing. AB - This study was done to analyze the effect of selenium on antioxidant status and expression of different connexins in diabetic wound healing. The levels of vascular endothelial growth factor, superoxide dismutase, lipid peroxide, and connexins were analyzed in wound tissues taken from diabetic and non-diabetic mice before and after sodium selenite administration. The mRNA transcript levels of Cx 26, 30.3, 31, 31.1, and 43 were significantly elevated in diabetic wounds as compared to the non-diabetic wounds. After selenium administration, the expression of connexins along with serum glucose decreases more significantly in diabetic wounds as compared to non-diabetic wounds. In diabetic wounds, the low levels of vascular endothelial growth factor and extracellular superoxide dismutase were restored to normal level following selenium administration. The lipid peroxidation decreased significantly in diabetic mice post-selenium administration. The histopathological analysis revealed that administration of selenium improves angiogenesis at the wound site. The results of this study demonstrate that selenium, acting as an essential component of the antioxidant system, normalizes the antioxidant status, and as an insulin mimetic compound, downregulates connexin expressions and induces angiogenesis. Together, these effects of selenium accelerate wound healing in diabetic conditions. PMID- 21633836 TI - Evaluation of molecular prognostic and predictive factors: an important step towards personalised treatment in non small cell lung cancer. AB - Treatment for patients with advanced NSCLC generally consists of chemotherapy, but response rates are modest and recurrence occurs for most patients after standard first-line platinum-based doublet therapy. Tailoring therapy to individual patient according to certain prognostic and predictive factors has the potential to improve outcome in NSCLC. This review focuses on the most important molecular prognostic and/or predictive factors in the treatment of advanced NSCLC; considering these molecular features, we also suggest a molecular-based treatment algorithm. PMID- 21633837 TI - Treatment of lupus: impact on quality of life. AB - Systemic lupus erythematosus (SLE) is an autoimmune disease that disproportionately affects women of childbearing age during their most productive years. Current therapeutic measures have improved patient survival; however, the impact of interventions on general and specific domains of health-related quality of life requires further study. Medical Outcomes Survey Short Form 36 (SF-36), the most commonly used measure, has been included in some SLE randomized controlled trials, but the observed effect sizes were generally small and in some cases negligible. An SLE patient's quality of life is known to be significantly worse than that of someone in the general population and perhaps worse than those with most other common chronic diseases. SF-36, although useful as a general measure, may not be the most sensitive way to gauge changes perceived by patients with SLE. Ongoing trials and observational longitudinal studies using lupus specific health-related quality-of-life measures may help better determine health related quality-of-life responses and determine the domains most amenable to interventions. PMID- 21633838 TI - Palytoxin in seafood by liquid chromatography tandem mass spectrometry: investigation of extraction efficiency and matrix effect. AB - Blooms of Ostreopsis spp. have been recently reported along the Mediterranean coasts of Spain, France, Italy, and Greece posing serious risks to human health. Occurrence of Ostreopsis spp. may result in palytoxin contamination of seafood and, in order to prevent sanitary risks, the need exists to develop efficient extraction procedures to be coupled to rapid and sensitive monitoring methods of palytoxin-like compounds in seafood. In the present study, the best conditions for both extraction of palytoxin from seafood and palytoxin quantification by using liquid chromatography tandem mass spectrometry (LC-MS/MS) were investigated. Three seafood matrices (mussels, sea-urchins, and anchovies) were selected and five different extraction systems were tested, namely: the official protocol for extraction of lipophilic toxins and various aqueous methanol or acetonitrile solutions (MeOH/H(2)O 1:1, MeOH/H(2)O 8:2, MeCN/H(2)O 8:2 and MeOH 100%). Extraction with MeOH/H(2)O 8:2 provided the best results in terms of accuracy and matrix interference on LC-MS/MS detection of palytoxin. Accuracy and intra-day reproducibility (n = 3) were evaluated for all the selected matrices but only for mussels at three spiking concentration levels, including the provisional limit proposed by the Community Reference Laboratory for marine biotoxins (250 MUg kg(-1)). Limits of quantitation of palytoxin in mussels, sea urchins and anchovies tissues were calculated using matrix-matched standards; taking into account extraction efficiency of MeOH/H(2)O 8:2, they resulted to be 228, 343, and 500 MUg kg(-1), respectively. PMID- 21633839 TI - Carbon nanotubes/pentacyaneferrate-modified chitosan nanocomposites platforms for reagentless glucose biosensing. AB - The design, characterization and applicability of a nanostructured biosensor platform are described. The biosensor is developed through the immobilization of three components: a polymeric chitosan network previously modified with a redox mediator (denoted as PCF-Pyr-Ch), an enzyme (glucose oxidase, chosen as a model) and carbon nanotubes onto a solid glassy carbon electrode (C). In order to assess the influence of the nanomaterial in the performance of the resulting analytical device, a second biosensor, free of carbon nanotubes, is developed. The characterization of both biosensing platforms was performed in aqueous phosphate buffer solutions using atomic force microscopy technique. In the presence of glucose, both systems exhibit a clear electrocatalytic activity, and glucose could be amperometrically determined at +0.35 V versus Ag/AgCl. The performance of both biosensors was evaluated in terms of sensitivity, detection limit and linear response range. Finally, the enhancement of the analytical response induced by the presence of carbon nanotubes was evaluated. PMID- 21633840 TI - Simultaneous determination of albendazole and its metabolites in fish muscle tissue by stable isotope dilution ultra-performance liquid chromatography tandem mass spectrometry. AB - A rapid, specific, and sensitive method utilizing ultra-performance liquid chromatography tandem mass spectrometry was developed and validated to determine albendazole, albendazole sulfoxide, albendazole sulfone, and albendazole 2 aminosulfone in fish muscle tissue. The fish samples were extracted with ethyl acetate, then the organic phase was evaporated to dryness, and the residue was reconstituted in methanol-water solution and cleaned up by n-hexane. Reversed phase separation of target compounds was achieved using a BEH C18 column and a gradient consisting of 0.2% (v/v) formic acid and methanol. Tandem mass spectrometry analyses were performed on a triple-quadrupole tandem mass spectrometer. In the whole procedure, the isotope-labeled internal standards were used to correct the matrix effect and variations associated with the analysis. The method was validated with respect to linearity, specificity, accuracy, and precision. The method exhibited a linear response from 0.1 to 20 ng mL(-1) (r(2) > 0.9985). The limit of quantitation for albendazole (ABZ), albendazole sulfoxide (ABZSO), albendazole sulfone (ABZSO(2)), and albendazole 2-aminosulfone (ABZ-2 NH(2)SO(2)) was 0.1, 0.1, 0.1, and 0.2 ng g(-1), respectively. The mean recoveries of ABZ, ABZSO, ABZSO(2), and ABZ-2-NH(2)SO(2) spiked at a level of 0.2 5.0 ng g(-1) were 95.3-113.7%, and the relative standard deviations of intra- and inter-day measurements were less than 6.38%. The method was later successfully applied to the determination of albendazole and its three metabolites in 60 fish samples collected from local markets. PMID- 21633841 TI - A routine accredited method for the analysis of polychlorinated biphenyls, organochlorine pesticides, chlorobenzenes and screening of other halogenated organics in soil, sediment and sludge by GCxGC-MUECD. AB - The analysis of persistent organic pollutants is a real challenge due to the large number of compounds with varying chemical and physical properties. Gas chromatography with electron capture detection or mass spectrometry has been the method of choice for the past 50 years. Comprehensive two-dimensional gas chromatography (GCxGC) coupled with micro-electron capture detector (MUECD) is a new method that can analyze polychlorinated biphenyls (PCBs), organochlorine pesticides (OCs) and chlorobenzenes (CBz) in a single analytical run with enhanced selectivity and sensitivity over single column methods and can also be used to screen for other halogenated organics in environmental samples. An accredited routine method using commercially available LECO GCxGC-MUECD and a column combination DB-1 * Rtx-PCB has been developed to analyse PCBs/OCs/CBz in soils, sediments and sludges. The method provides quantification of Aroclors and Aroclor mixtures to within 15% of target values and sub-nanogrammes per gramme detection limits. PMID- 21633842 TI - Effect of D-allose on prostate cancer cell lines: phospholipid profiling by nanoflow liquid chromatography-tandem mass spectrometry. AB - D-Allose, a rare, naturally occurring monosaccharide, is known to exert anti proliferative effects on cancer cells. The effects of D-allose on the cellular membranes of hormone-refractory prostate cancer cell line (DU145), hormone sensitive prostate cancer cell line (LNCaP), and normal prostate epithelial cells (PrEC) were studied at the molecular level by phospholipid (PL) profiling using a shotgun lipidomic method. The molecular structures of 85 PL species including 23 phosphatidylcholines, 12 phosphatidylethanolamines (PEs), 11 phosphatidylserines (PSs), 16 phosphatidylinositols, 9 phosphatidic acids (PAs), and 14 phosphatidylglycerols (PGs) were identified by data-dependent collision-induced dissociation of nanoflow liquid chromatography-tandem mass spectrometry, and the PL amounts were quantified. The addition of D-allose to prostate cancer cell lines during their growth phases had negligible or decreased effects on the relative regulation of PL species, but several new PS molecules (two for DU145 and three for LNCaP) emerged. In contrast, experiments on the PrEC cell line revealed that some high abundant species (14:0/14:0-PE, 16:2/16:0-PG, and 20:6/18:1-PA) showed significant increases in concentration. These findings support a mechanism for the anti-proliferative effect of D-allose on prostate cancer cell lines that involves the induction of programmed cell death since PS molecules are known to induce apoptosis. Principal component analysis was carried out to examine differences in PL distributions among the three cell lines promoted by D-allose. PMID- 21633843 TI - Seasonality of gregarine parasitism in the damselfly, Nehalennia irene: understanding unimodal patterns. AB - We studied parasitism by gut protozoans (Apicomplexa: Eugregarinidae) in the damselfly, Nehalennia irene (Hagen) (Odonata: Coenagrionidae). We tested whether there was any seasonal pattern, as has been found for other parasites of damselflies and which has implications for selection on emergence and breeding. Using aggregate data from 12 date-by-site comparisons involving five sites, we found that both prevalence and intensity of gregarine parasitism were seasonally unimodal. Parasitism first increased and then declined seasonally after peaking mid-season. This damselfly species has shown seasonal increases in density followed by declines at several sites including a site sampled in this study. Therefore, similar seasonal changes in a directly transmitted parasite were expected and are now confirmed. Other factors that might account for seasonal changes in parasitism by gregarines are either unlikely or can be discounted including sampling of older damselflies mid-season but not late in the season, or sex biases in parasitism and overrepresentation of the more parasitized sex mid season. PMID- 21633844 TI - Ultrastructure, chromosomal karyotype, and molecular phylogeny of a new isolate of microsporidian Vairimorpha sp. BM (Microsporidia, Nosematidae) from Bombyx mori in China. AB - The spore morphology, chromosomal karyotype, and molecular systematic of a new microsporidian which was isolated from the domesticated silkworm Bombyx mori (Lepidoptera: Bombycidae) in Shandong, China have been studied. The spores were long oval and measured 3.4 * 1.6 MUm on fresh smears. Ultrastructure of the spores was characteristic for the genus Vairimorpha: 13-15 polar filament coils, posterior vacuole, and a diplokaryon. Six chromosome bands have been separated by pulsed field gel electrophoresis. The sequenced complete rRNA gene of this isolate is 4,231 bp long. Phylogenetic analysis based on SSU rRNA gene and LSU rRNA gene both revealed that this novel microsporidian which was isolated from B. mori had close relationship to the genus Vairimorpha, not to the genus Nosema. Moreover, the organization of the rRNA units of this microsporidian is not similar to that of Nosema bombycis, but same to that of other microsporidian, such as Vairimorpha necatrix. Although this microsporidian, designed as Vairimorpha sp. BM, was isolated from B. mori, all of these informations indicate that this isolate is closely related to the Vairimorpha group. PMID- 21633845 TI - Structure and dynamics of microbial community in full-scale activated sludge reactors. AB - Phospholipid fatty acid (PLFA) profiles in four full-scale activated sludge reactors (ASR1 ~ 4) treating municipal wastewater, South Korea, were monitored to evaluate the influence of influent water quality on microbial community structure (MCS) and the effect of the MCS on effluent water quality. In ASR1 ~ 3, PLFA profiles were very similar, regardless of the influent water quality and seasonal differences, and 16:17c/15:0iso2OH and 16:0 were dominant. PLFA profiles in ASR4 during summer and autumn were very similar to those in ASR1 ~ 3, but increases in specific fatty acids, 16:1omega5c, 11methyl18:1omega7c and 15:0iso3OH, were found in ASR4 during winter and spring, with relatively high total suspended solid (TSS) concentrations in the effluent. 16:1omega5c and 15:0iso3OH, possibly related with Flexibacter sp., caused a bulking problem in the activated sludge. The community diversity indices such as Shannon diversity and equability decreased in summer but increased in autumn in all the ASRs. Canonical correspondence analysis results suggested that the influent BOD concentration played the most important role in changing MCS, followed by influent TSS concentration. In addition, the TSS and total phosphorus concentrations in the effluent were significantly affected by the change of the MCS. PMID- 21633846 TI - Radiologic and nuclear medicine predictors of tumor invasiveness in patients with clinical stage IA lung adenocarcinoma. AB - BACKGROUND: The aim of this study was to analyze parameters to predict tumor invasiveness according to high-resolution computed tomography and positron emission tomography in patients with clinical stage IA lung adenocarcinoma. METHODS: A total of 122 patients with clinical stage IA lung adenocarcinoma were enrolled in the study. Receiver operating characteristic (ROC) curves were constructed for three factors--the degree of solid tumor component (solid%), maximum standard uptake value (SUVmax) and tumor size--and cutoff values were determined to reveal the highest sensitivity and specificity to diagnose tumor invasiveness. We created an algorithm for detecting tumor invasiveness (model 1). The data for the three factors were combined and their ROC curves constructed (model 2). A prospective study was conducted to validate the utility of these models. RESULTS: Multivariate analysis identified solid%, SUVmax, and tumor size as potentially important predictors of tumor invasiveness. In the ROC curve analysis, solid% (area under the curve was 0.882) had the largest area under the curve, followed by the SUV (0.867) and tumor size (0.747). The combination assay using all three factors had the highest sensitivity and specificity for prediction (0.902). Models 1 and 2 were applied to the prospectively enrolled cases, and their utility was reviewed. Both models showed 100% sensitivity, with model 2 showing a slightly higher diagnostic value than model 1. CONCLUSIONS: The solid portion ratio was a more powerful clinical predictor for lymphovascular invasion than the SUVmax. Our novel scoring model for tumor invasiveness can be employed for preoperative assessment of tumor invasiveness. PMID- 21633847 TI - Bilateral acute depigmentation of the iris first misdiagnosed as acute iridocyclitis. AB - Acute depigmentation of the iris is a new condition characterized by pigment dispersion in the anterior chamber, depigmentation of the iris stroma, and pigment deposition in the anterior chamber angle. A 33-year-old woman using a topical corticosteroid every 2 h for the treatment of acute iridocyclitis was referred to our clinic to seek another opinion because her symptoms had not improved. An ocular evaluation of the patient revealed pigment precipitates on the corneal endothelium, pigment dispersion in the anterior chamber, symmetrical diffuse depigmentation, granularity of the iris stroma, and pigment deposition in the trabecular meshwork. These findings suggested a diagnosis of bilateral acute depigmentation of the iris (BADI) instead of iridocyclitis. Clinicians should be careful in the differential diagnosis of iris depigmentation from iridocyclitis to avoid the unnecessary use of high-dose topical corticosteroids. PMID- 21633848 TI - Asymptomatic subconjunctival entrapment of a cilium. AB - We report an unusual case of asymptomatic subconjunctival entrapment of a cilium. A 64-year-old man presented with a 3 month history of a painless black line in the conjunctiva of the left eye, and slit-lamp examination revealed a cilium under the conjunctiva. The patient remained asymptomatic without conjunctival edema or hyperemia during a 2 week follow-up. To the best of our knowledge, this is the first report of an asymptomatic cilium inside the subconjunctival space. PMID- 21633849 TI - Diagnosis and treatment of subclinical hypothyroidism detected by neonatal screening. AB - BACKGROUND: This study was undertaken to explore the clinical outcome and prognosis of subclinical hypothyroidism detected by newborn screening. METHODS: Newborn screening was conducted at 1156 health care institutions in Zhejiang Province from October 1999 to September 2006. Included were (1) infants who had thyroid-stimulating hormone (TSH) >= 20 mU/L, and normal or lower normal levels of triiodothyronine (T(3)) and thyroxine (T(4)) and (2) infants with TSH between 5.6 mU/L and 20 mU/L at a confirmatory examination and follow-up showing TSH levels >= 20 mU/L or delayed reduction in T(4) levels. These infants were considered as having subclinical hypothyroidism and levothyroxine (L-T(4)) at an initial dose of 3-5 MUg/kg per day was administered. The levels of TSH and T(4), developmental quotient (DQ), and index of growth were evaluated. RESULTS: A total of 204 infants met our criteria for subclinical hypothyroidism, with an incidence of 1/8809. After 2-4 weeks of standard therapy, serum TSH level dropped to normal and T(4) reached a higher normal level in all the 204 infants. Evaluations of 60 patients after 2 years of therapy showed that their average DQ was 101 +/- 14.61, and body weight and height were within the normal ranges. Bone age test for 54 patients revealed normal development in 44, slightly retarded development in 7, and advanced development in 3. CONCLUSIONS: Newborns with high TSH levels should be given particular attention to ensure early diagnosis. A L-T(4) dose of 3-5 MUg/kg per day was effective in the initial treatment of subclinical hypothyroidism. PMID- 21633850 TI - Multiple calcified primary central nervous system lymphoma with immunodeficiency in a child. AB - BACKGROUND: Multiple calcified primary central nervous system lymphoma (PCNSL) is extremely rare in childhood. METHODS: We report a 4-year-old boy suffering from multiple calcified B-cell lymphoma in the brain with immunodeficiency. RESULTS: The boy had a history of walking weakness and seizure for 4 months. The serum levels of immunoglobulin G, A and M were decreased. Brain MRI showed multiple lesions which had ring enhancement. CT showed calcification in all of the lesions. The boy was firstly misdiagnosed with multiple chronic brain abscesses. Pathological analysis of biopsy confirmed the diagnosis of anaplastic diffuse large B-cell lymphoma. CONCLUSION: PCNSL should be included in the differential diagnosis of intracranial mass with calcification. PMID- 21633851 TI - Glucosidase acid beta gene mutations in Egyptian children with Gaucher disease and relation to disease phenotypes. AB - INTRODUCTION: More than 200 mutations have been found in patients with Gaucher disease (GD) and some mutations usually have a high frequency in certain populations. Genotype/phenotype correlation in patients with GD has not been established. This study was designed to determine underlying mutations in Egyptian children with GD and to assess their relation to disease phenotypes. METHODS: This study comprised 17 patients with GD and 10 healthy controls. Thirteen patients were type 1 GD, 2 type 2, and 2 type 3. DNA was extracted from peripheral blood leukocytes. Exons 9 and 10 were amplified by polymerase chain reaction, and deoxyribonucleic acid sequencing was done with an ABI 310 genetic analyzer. RESULTS: Wild type allele was detected in 95% (19/20) and a normal variant in 5% (1/20) of controls. L444P allele was encountered in 50% (13/26) of the alleles in type 1 patients, H451P in 7.7% (2/26) and recombinant alleles (RecNcil, RecNcil + M450L, RecFs, RecFs + M450L) in 34.6% (9/26). L444P and Rec alleles each occurred in 50% (2/4) of type 2 and 3 patients. A new mutation was seen in this study {g.7336A>C, (M450L)} and 2 mutant alleles were not determined. Type 1 GD patients had L444P/L444P genotype (23.1%) and Rec alleles/L444P (53.8%), while type 2 and 3 GD patients had Rec alleles/L444P genotypes (100%) with a poor phenotype/genotype correlation. CONCLUSIONS: L444P and Rec alleles are common in the studied patients. Novel mutations are continuously detected, adding to the expanding panel of GD mutations. No significant genotype-phenotype association was observed. PMID- 21633852 TI - Cytomegalovirus enterocolitis presenting as abdominal compartment syndrome in a premature neonate. AB - BACKGROUND: Cytomegalovirus (CMV) enterocolitis is an uncommon intestinal disorder of newborns that is often initially misdiagnosed as necrotizing enterocolitis. METHODS: We treated a premature twin boy with CMV enterocolitis who presented with abdominal compartment syndrome requiring urgent decompression. All patients with neonatal CMV enterocolitis reported were reviewed. RESULTS: Nine previously reported patients with neonatal CMV enterocolitis presented with abdominal distention and signs of sepsis. At the time of surgery, either perforation or stricture was identified. The current report is the first to present with clinical signs of abdominal compartment syndrome. CONCLUSION: CMV is a rare cause of neonatal enterocolitis. Surgical intervention is required for bowel perforation, stricture, or abdominal compartment syndrome. PMID- 21633853 TI - Non-Hodgkin lymphoma in childhood and adolescence: frequency and distribution of immunomorphological types from a tertiary care center in South India. AB - BACKGROUND: There is a dearth of published literature on the frequency and distribution of pediatric and adolescent non-Hodgkin lymphoma (NHL) in India according to the 2001 WHO classification. The aim of this study was to record the distribution of the different subtypes, analyze the major subtypes, and compare it with the published data from other countries. A review of pediatric NHL statistics from population-based cancer registries was included in this study. METHODS: The study was conducted using information retrieved from the files of our institution. A total of 467 patients with lymphoma (excluding mycosis fungoides) were recorded in the under 20 years group over a period of 6 years, of which 252 patients suffered from NHL. The demographic characteristics, frequency and distribution of different subtypes were noted and compared with published reports from other parts of the world. RESULTS: T-lymphoblastic lymphoma/leukemia constituted the majority (32.1%) of all NHLs in children and adolescents in our study. The other major subgroups were Burkitt's lymphoma, anaplastic large cell lymphoma, and diffuse large B cell lymphoma. Burkitt's lymphoma in this study had clinical presentations similar to those seen in western countries. CONCLUSIONS: The distribution of different subtypes of lymphoma in pediatric and adolescent NHL in India differs considerably from that in western countries and other eastern countries. PMID- 21633854 TI - Association between asthma and dental caries in the primary dentition of Mexican children. AB - BACKGROUND: Asthma has been associated with a great number of negative health outcomes. This study was undertaken to detect the association between asthma and dental caries in the primary dentition. METHODS: Data were obtained from a cohort of 1160 Mexican children aged 4-5 years. We used the questionnaire of the International Study of Asthma and Allergies Diseases in Childhood. Asthma was classified according to parents' reports. Caries indexes were measured as the number of decayed (d), missing (m), and filled (f), teeth (t) (dmft) or surfaces (dmfs). Decayed teeth included initial caries in this study. Adjusted odds ratios (adjusted ORs) were determined for asthma using logistic regression model. Gender, sugary products consumption, and oral hygiene habits were utilized as covariates. RESULTS: The prevalence of dental caries was 17.9% in the 1160 children. Approximately 226 (19.5%) children were identified with asthma. Among them, 166 (73.5%) presented with symptoms during the day and 60 (26.5%) during the day and night. The prevalence of caries in children with asthma was 19.9%; it was higher in children with nocturnal asthma symptoms than in those with asthma symptoms only during the day. Logistic regression model showed that asthma (adjusted OR=1.24; 95% confidence interval [95% CI]=0.84-1.81) was not associated with caries. A significant association was found between nocturnal asthma symptoms (adjusted OR=1.85; 95% CI=1.00-3.44) and dental caries. CONCLUSIONS: Asthma is not associated with dental caries. Nocturnal asthma symptoms appear to be associated with dental caries in the primary dentition. PMID- 21633855 TI - A novel heterozygous NR1H4 termination codon mutation in idiopathic infantile cholestasis. AB - BACKGROUND: This study aimed to evaluate the genetic effect of the NR1H4 gene in the pathogenesis of idiopathic infantile cholestasis of Chinese subjects in Guangxi, China. METHODS: Seventy-eight patients with idiopathic infantile cholestasis served as a study group and 95 infants without cholestasis as controls. Genomic DNA was extracted from peripheral venous blood leucocytes by phenol chloroform procedures. Polymerase chain reaction (PCR) was used to amplify all coded exons of NR1H4, and single-strand conformation polymorphism (SSCP) was used to analyze all amplification fragments. The PCR products with abnormal bands in SSCP were sequenced using an ABI 3100 sequencer. RESULTS: A novel heterozygous termination codon mutation in NR1H4 exon 5 (NR1H4 R176X, CGA-TGA) was found in one of the 78 patients. The patient with mutation R176X had high levels of bilirubin, alanine aminotransferase, gamma-glutamyltransferase, cirrhosis and ascites despite biliary tract flushing procedures and drug therapy. In the other patients and controls, no mutation was detected. CONCLUSIONS: Heterozygous termination codon mutation of NR1H4 R176X was found in idiopathic infantile cholestasis. The novel mutation is useful to establish particular characteristics for differential diagnosis of idiopathic infantile cholestasis and to determine the influence of such gene defects in the prognosis. PMID- 21633856 TI - The course of early-onset multiple sclerosis in Iraqi children. AB - BACKGROUND: Multiple sclerosis (MS) is a chronic demyelinating disease of the central nervous system that is increasingly recognized worldwide in children and adolescents. The current study aimed at identifying the clinical characteristics of MS with onset under 18 years of age. METHODS: This cross-sectional study was conducted in the Multiple Sclerosis Center archive system in Baghdad Teaching Hospital during the period from March 1 to May 15, 2008. The records of 1125 MS patients from 2000 to 2008 were reviewed. Among them 77 patients had the onset of MS under 18 years of age. RESULTS: Two thirds of the patients were female (a female/male ratio of 1.6:1). The mean age of the patients at the onset of the disease was 14.95 +/- 3.21 years, and the mean time between the first and second attacks was 3.06 +/- 4.09 years. Seventy patients (90.9%) had an initial course of relapse remitting MS. Among them 9 (12.9%) progressed to secondary progressive MS after a mean duration of 9.87 +/- 4.14 years. The remaining 7 patients had primary progressive MS associated with optic neuritis and brain stem lesion. Fifty-nine (76.6%) patients had monofocal signs and 18 (23.4%) had polyfocal signs. The mean extended disability status scale score was 4.15 +/- 2.17 and the mean progression index was 1.44 +/- 2.31. There was a strong inversed correlation between the progression index and interval between the first and second attacks (P=0.0001). CONCLUSIONS: The results of the present study show that the course of MS in Iraqi children and adolescents is more aggressive than in children from other countries. This finding needs to be evaluated by further studies. PMID- 21633857 TI - What is the result: chylous leakage following extensive radical surgery of neuroblastoma. AB - BACKGROUND: Postoperative chylous leakage is a rare complication that results from disruption of either mediastinal or retroperitoneal lymphatic channels during dissection or from obstruction by original lesions such as a malignancy. There is lack of clinical experience in pediatric patients about how to manage the leakage and what the result will be. METHODS: We retrospectively analyzed the clinical outcomes of 5 children with neuroblastoma (NB) (stage 4 in 4 children and stage 1-2 in 1 child) who had received non-surgical treatment of chylothorax and/or chylous ascites after retroperitoneal/posterior mediastinal extensive radical resection of NB for complete tumor removal. Conservative therapy with low fat diet, medium-chain triglyceride and/or total parenteral nutrition was the mainstay treatment for chylous leakage. RESULTS: Four of the 5 children recovered after 6-32 days of conservative treatment, and the last one who did not respond was cured by surgical management for chylous fistula 45 days after primary surgery. Postoperative imaging showed that more than 90% of tumors were resected and all of them showed very good partial remission (VGPR). Among the 4 patients in stage 4, 3 relapsed after radical resection of NB. The patient of stage 1-2 was still in VGPR. CONCLUSIONS: The majority of patients with chylous ascites/chylothorax after extensive radical surgery for posterior mediastinum/retroperitoneum NB could be cured by non-surgical treatment. But the final result of original disease has not been greatly changed by intensive surgery for stage 4 NB. PMID- 21633858 TI - Familial hypomagnesemia with hypercalciuria and nephrocalcinosis in three siblings having the same genetic lesion but different clinical presentations. AB - BACKGROUND: This article summarizes the varying clinical manifestations of three siblings with familial hypomagnesemia with hypercalciuria and nephrocalcinosis (FHHNC) caused by the same genetic lesion. METHODS: The medical records of three siblings with FHHNC (one girl and two boys, aged 6 to 12 years) were reviewed and the clinical manifestations and treatment of their disease were described. RESULTS: Despite varying phenotypes, each sibling had the same genetic lesion-a novel homozygous mutation in CLDN16 (c.211A>G, M71V). CONCLUSION: Although FHHNC is a rare disorder, this report is significant for the following reasons: (i) it describes a novel CLDN16 mutation causing FHHNC, adding to the literature of FHHNC-causing CLDN16 mutations; (ii) it suggests that genes other than CLDN16 or epigenetic factors are involved in the clinical spectrum of FHHNC; and (iii) it reinforces the variability of disease manifestation and genotype-phenotype correlations. PMID- 21633859 TI - Follow-up results of children with melamine induced urolithiasis: a prospective observational cohort study. AB - BACKGROUND: Melamine-contaminated milk powder was the cause of the 2008 outbreak of urolithiasis in young children and infants in China, but the prognosis of these children remains unknown. We hypothesized that urolithiasis induced by melamine-contaminated milk powder may be associated with secondary renal injury. METHODS: A total of 8335 children (<=6 years old) with a history of consuming melamine-contaminated milk powder were screened. Urine analysis and urinary system ultrasonography were performed. For children with urolithiasis, the basic information and the results of examination were recorded, and effective therapy was given. They were followed up for 6 months after the original diagnosis, and urinary microprotein profiles were measured. RESULTS: Of the 8335 children, 105 (1.26%) were diagnosed with melamine-contaminated milk powder-associated urolithiasis. The size of the stone was correlated with the duration of exposure to melamine. Six months later, 69.8% (67) of the children with urolithiasis passed stones (follow-up rate: 91.4%). Of the 67 children, 28 passed stones within 2 months. The higher possibility of passing a stone was correlated with the smaller diameter of the stone (P<0.001). The detection rate of abnormal urinary microprotein excretion (microalbumin, immunoglobulin G, and N-acetyl-beta D-glucosidase) was 52.4% in children with persistent stones and 38.2% in those who passed their stones. The detection rate was lower in children who passed stones within 2 months (31.8%) than in those who passed stones in 2 to 6 months (50.0%). The levels of microalbumin/creatinine and immunoglobulin G/creatinine were significantly higher in children with persistent stones than in those who passed their stones. CONCLUSIONS: Early passage of a stone may reduce the renal injury induced by melamine-contaminated milk powder-associated urolithiasis. PMID- 21633860 TI - Are the neonatal outcomes similar in large-for-gestational age infants delivered by women with or without gestational diabetes mellitus? AB - BACKGROUND: Infants are considered large for gestational age (LGA) if their birth weight is greater than the 90th percentile for gestational age and they have an increased risk for adverse perinatal outcomes. Maternal diabetes is one of the factors affecting birthweight. However there are limited data on the perinatal outcomes of infants of gestational diabetic mothers. The aim of the present study was to compare the neonatal outcomes of LGA infants delivered by women with and without gestational diabetes mellitus. METHODS: This was a retrospective study of LGA infants of >=36 weeks of gestation born at the Gazi University Medical School Hospital during the period of 2006-2009. Neonatal outcomes included hypoglycemia and polycythemia in the early neonatal period and hospital admissions. The Chi square and Student's t test were used for comparing variables. RESULTS: Seven hundred eligible infant-mother pairs were enrolled in the study. Eighty-seven of them (12.4%) were infants of gestational diabetic mothers and 613 (87.6%) were infants of non-diabetic mothers. The incidence of hypoglycemia at the first hour was higher in infants of diabetic mothers (12.8%) than in infants of non-diabetic mothers (5.3%) (P=0.014). Polycythemia was also more frequently observed in infants of the gestational diabetic mothers (9.3%) than in infants of the non diabetic mothers (3.0%) (P=0.010). Although overall hospital admission rates were not different between the two groups, infants of diabetic mothers were more likely to be admitted because of resistant hypoglycemia (P=0.045). CONCLUSIONS: The results of this study suggested that LGA infants of mothers with gestational diabetes mellitus were at a greater risk for hypoglycemia and polycythemia in the early neonatal period than LGA infants of nondiabetic mothers. PMID- 21633861 TI - Hereditary tyrosinemia type 1 from a single center in Egypt: clinical study of 22 cases. AB - BACKGROUND: Hereditary tyrosinemia type 1 (HT1) is an increasingly recognized inborn error of metabolism among Egyptian children. This study was undertaken to define the presenting clinical, biochemical and imaging features and outcome of 2 (2-motrp-4-trifluoromethylbenzoyl)-1, 3-cyclohexanedione (NTBC) therapy and liver transplantation in a cohort of Egyptian children diagnosed with HT1. METHODS: The study was carried out at the Pediatric Hepatology Unit at Cairo University Children's Hospital. HT1 was diagnosed by quantification of succinylacetone (SA) in dry blood spots. RESULTS: Twenty-two patients were diagnosed with HT1 in a period of 3 years from August 2006 to July 2009. Infants with focal hepatic lesions and hepatomegaly (n=13) were younger at diagnosis than those with rickets (n=5) (median age: 3.25 vs. 10 months; P=0.05). Alpha fetoprotein was highly elevated in all children. Seven children died within a few weeks of diagnosis before therapy was initiated. Ten children were treated with NTBC. The response to NTBC treatment was apparent by a steep drop in serum alpha fetoprotein (AFP) and undetectable SA in urine within 2 months. Three children underwent living donor liver transplantation after treatment with NTBC for 10, 18 and 22 months respectively, despite adequate response to therapy because of financial issues. The explanted livers were all cirrhotic with no dysplasia or malignant transformation. CONCLUSIONS: Focal hepatic lesions are the commonest presentation of HT1 patients and they present at an earlier age than rickets. NTBC is effective but very expensive. Liver transplantation is still considered in HT1 patients. PMID- 21633862 TI - Niemann-Pick disease type C: analysis of 7 patients. AB - BACKGROUND: Niemann-Pick disease type C (NP-C), derived from mutation of the NPC1 or NPC2 gene, is one of the recessive lysosomal lipid storage disorders that are difficult to diagnose and treat. Since NP-C has been rarely reported in China, we reviewed 7 patients with NP-C. METHODS: The 7 patients had been diagnosed with NP C from 2007 to 2010 at our department and their laboratory and clinical data were analyzed. RESULTS: The 7 patients, 5 males and 2 females, included 4 patients of late infantile subtype and 3 patients of juvenile subtype, in which patients 2 and 3 were siblings. Their clinical symptoms occurred from 4 to 10 years of age, exhibiting as progressive cognitive and language impairment as well as motor retrogression. Six patients were caught by focal or generalized seizures from 1 to 4 years after the onset of the disease. Vertical supranuclear gaze palsy, dysarthria, dysphagia, internal rotation and adduction of bilateral hands and splenomegaly occurred following the progress of clinical symptoms. Five patients had laughter-cataplexy. MRI showed mild brain atrophy in 6 patients. Reduction of total cholesterol, high density lipoprotein cholesterol and low density lipoprotein cholesterol occurred in 6 patients. Sea-blue cells and Niemann-Pick cells were found in bone marrow smears. The activity of acid sphingomyelin enzyme was normal or only slightly lower. Supporting or symptomatic treatment improved common clinical symptoms. CONCLUSIONS: NP-C is a rare autosomal recessive inherited lysosomal storage disease that affects the intellectual development of children and may lead to dementia, vegetative state or death. Clinical features of this disease include vertical supranuclear gaze palsy, seizures and cataplexy. Laboratory features include abnormal plasma cholesterol level, and sea-blue cells and Niemann-Pick cells in bone marrow smears. The treatments of the disease include supporting or symptomatic administration. PMID- 21633863 TI - Editors' overview: moral responsibility in technology and engineering. PMID- 21633864 TI - Design, development, and evaluation of a second generation interactive Simulator for Engineering Ethics Education (SEEE2). AB - This paper describes a second generation Simulator for Engineering Ethics Education. Details describing the first generation activities of this overall effort are published in Chung and Alfred (Sci Eng Ethics 15:189-199, 2009). The second generation research effort represents a major development in the interactive simulator educational approach. As with the first generation effort, the simulator places students in first person perspective scenarios involving different types of ethical situations. Students must still gather data, assess the situation, and make decisions. The approach still requires students to develop their own ability to identify and respond to ethical engineering situations. However, were as, the generation one effort involved the use of a dogmatic model based on National Society of Professional Engineers' Code of Ethics, the new generation two model is based on a mathematical model of the actual experiences of engineers involved in ethical situations. This approach also allows the use of feedback in the form of decision effectiveness and professional career impact. Statistical comparisons indicate a 59 percent increase in overall knowledge and a 19 percent improvement in teaching effectiveness over an Internet Engineering Ethics resource based approach. PMID- 21633865 TI - Defining viability in mammalian cell cultures. AB - A large number of assays are available to monitor viability in mammalian cell cultures with most defining loss of viability as a loss of plasma membrane integrity, a characteristic of necrotic cell death. However, the majority of cultured cells die by apoptosis and early apoptotic cells, although non-viable, maintain an intact plasma membrane and are thus ignored. Here we measure the viability of cultures of a number of common mammalian cell lines by assays that measure membrane integrity (a measure of necrotic cell death) and assays that measure apoptotic cells, and show that discrepancies in the measurement of culture viability have a significant impact on the calculation of cell culture parameters and lead to skewed experimental data. PMID- 21633866 TI - Pleiotropic effects of a schweinfurthin on isoprenoid homeostasis. AB - The schweinfurthins, a family of natural products derived from the isoprenoid biosynthetic pathway (IBP), have marked growth inhibitory activity. However, the biochemical basis for the schweinfurthins cellular effects has remained ill defined. Here, the effects of the synthetic schweinfurthin, 3-deoxyschweinfurthin (3dSB) on multiple aspects of isoprenoid homeostasis are explored. Cytotoxicity assays demonstrate a synergistic interaction between 3dSB and the HMG-CoA reductase inhibitor lovastatin but not with other IBP inhibitors in a variety of human cancer cell lines. The cytotoxic effects of 3dSB were enhanced in cells incubated in lipid-depleted serum. 3dSB was found to enhance the lovastatin induced decrease in protein prenylation. In addition, 3dSB decreases intracellular farnesyl pyrophosphate and geranylgeranyl pyrophosphate levels in both established cell lines and primary cells. To determine whether 3dSB alters the regulation of expression of genes involved in isoprenoid homeostasis, real time PCR studies were performed in human cell lines cultured in either lipid replete or -deplete conditions. These studies demonstrate that 3dSB abrogates lovastatin-induced upregulation of sterol regulatory element-containing genes and lovastatin-induced downregulation of ABCA1. In aggregate, these studies are the first to demonstrate that a schweinfurthin exerts pleiotropic effects on isoprenoid homeostasis. PMID- 21633867 TI - Risk factor for axillary lymph node metastases in microinvasive breast cancer. AB - BACKGROUND: Sentinel lymph node biopsy (SLNB) in patients with ductal carcinoma in situ (DCIS) was controversial. Usually we did not do a SLN biopsy when we performed conserving operations with small-sized DCIS. However, sometimes we find DCIS with microinvasive breast cancer (MIC) after the operation. Must reoperations be performed in all patients? The incidence of axillary metastases in microinvasive breast cancer (MIC) has not been extensively studied. We determined the incidence of positive axillary lymph node (ALN) in patients with MIC and the predictive factors of ALN metastases in these patients. METHODS: Between July 1989 and December 2008, 9635 patients had operation on invasive breast cancer in Asan Medical Center. Among these patients, 319 patients had MIC. The research conducted on the 293 patients (excluded were 26 who did not receive axillary lymph node dissection or SLN biopsy). We retrospectively checked clinical and pathologic variables. RESULTS: There were 22 cases of ALN metastases identified in this group of patients (7.5%). Lymphatic invasion (P < .001) and positive estrogen receptor status (P = 03) were independent significant predictors of axillary metastases. CONCLUSIONS: Microinvasive breast cancer is associated with a low rate of lymph node metastases. Some breast cancer patients with MIC at low likelihood of lymph node metastases may be spared lymph node evaluation. PMID- 21633868 TI - Diagnostic accuracy of surgeon-performed ultrasound-guided fine-needle aspiration of thyroid nodules. AB - BACKGROUND: There is scant data concerning surgeon-performed thyroid fine-needle aspiration (FNA), and controversy regarding its accuracy in larger nodules. This study aimed to specifically assess accuracy of surgeon-performed ultrasound (US) guided FNA on a per-nodule basis, with a subanalysis of nodule size. METHODS: Data of 1,000 surgeon-performed US-guided thyroid FNAs at a single institution from 2000 to 2010 were prospectively collected. Standard clinical information, FNA results using the Bethesda criteria, and final histology were recorded. RESULTS: Fine-needle aspiration results were reported as: cancer (7%), suspicious for cancer (2%), suspicious for follicular neoplasm (17%), atypia of unknown significance (AUS) (1%), benign (67%), and insufficient (6%). Of nodules with FNA results of cancer, suspicious for cancer, suspicious for follicular neoplasm, and atypia of unknown significance, 94% were operated on, with malignancy rates of 97%, 58%, 21%, and 12%, respectively. Of nodules with benign FNA, 26% underwent surgery for associated symptoms, concerning features, or other remote pathology. A total of 56% were followed, and 18% were lost to follow-up. Of nodules with insufficient FNA, 46% had repeat FNA (yielding a diagnosis in 81%), 23% underwent surgery, 21% with hypocellular features were followed, and 9% were lost to follow up. In size subanalysis, there was no statistically significant difference in risk of malignancy or increased rate of falsely negative FNA with increasing nodule size. CONCLUSIONS: The Bethesda system appropriately stratified lesions for risk of malignancy, and repeat FNA had high diagnostic yield in lesions with inadequate FNA. The results suggest no trend toward larger lesions harboring thyroid malignancy nor an increased likelihood of false-negative benign FNA. PMID- 21633869 TI - Prevention, evaluation, and treatment of leaks after gastrointestinal surgery : prevention of leaks after pancreatic surgery. AB - Pancreatic fistula remains a frequent and serious complication following pancreatic surgery. The incidence ranges from 10-20% in most series, with pancreatic texture being the most common predictor of the complication. There is little level 1 evidence to support any potential measures to reduce the incidence of pancreatic fistula. PMID- 21633870 TI - Prolonged transcranial Doppler monitoring after aneurysmal subarachnoid hemorrhage fails to adequately predict ischemic risk. AB - BACKGROUND: Stroke is common after aneurysmal subarachnoid hemorrhage (aSAH). Transcranial Doppler ultrasound (TCD) monitoring is often employed to identify vasospasm and allow intervention to avoid infarction. The required duration of monitoring has not been established. We aim to determine if 10 days of TCD monitoring identifies all patients at risk for infarction. METHODS: A 3 year retrospective analysis of aSAH patients admitted to a neurovascular center was undertaken. Eligible patients were aged 18-85 years, presenting within 2 days of hemorrhage who had underwent TCD monitoring through post bleed day 10. Patients were assessed to determine if vasospasm onset occurred after 10 days with resulting stroke. Assessment of variables potentially impacting vasospasm onset and infarction were completed. RESULTS: 107 patients met criteria with 51 (48%) demonstrating vasospasm and 31 (29%) developing stroke. Of those suffering stroke, 22 (71%) demonstrated vasospasm while 9 (22%) did not. Two (2%) patients developed vasospasm only after day 10, neither experiencing stroke. Time to vasospasm onset (5.5 +/- 2.5 days) was not impacted by common radiologic or clinical scales. Glasgow Coma Scale (GCS), Hunt and Hess Score (H&H), WFNS, ventriculostomy placement, intubation, and intraventricular hemorrhage were associated with likelihood of stroke (P < 0.05). The negative predictive value of TCD for identifying stroke risk was 84% while the sensitivity was 71%. CONCLUSIONS: TCD identification of vasospasm after day 10 is rare. Stroke is more likely to result from poor detection than from brevity of TCD monitoring. Improved or alternative monitoring is needed to effectively identify ischemia and prevent stroke. PMID- 21633871 TI - Attending code status discussions at admission. PMID- 21633872 TI - Successful treatment of acute respiratory distress syndrome after hysterectomy for life-threatening atonic bleeding by inhaled nitric oxide. AB - We report a case of a 33-year-old female who developed severe acute respiratory distress syndrome (ARDS) after emergency hysterectomy for life-threatening atonic bleeding. A marked decline in pulmonary oxygenation was observed during the surgery, which led to a diagnosis of ARDS. Following admission to the intensive care unit, hypoxia became critical, with a PaO(2)/F(I)O(2) value of 52 even after recruitment maneuvers. Inhaled nitric oxide (NO 10 ppm) was administered to the patient as a rescue treatment, resulting in a gradual but dramatic improvement in pulmonary oxygenation. Although several randomized trials have failed to confirm the beneficial effects of NO on morbidity in patients with ARDS, NO administration is worth consideration as treatment prior to invasive treatments, such as extracorporeal membrane oxygenation, for patients with acute lung injury/ARDS. PMID- 21633873 TI - Human brain activity associated with painful mechanical stimulation to muscle and bone. AB - PURPOSE: The purpose of this study was to elucidate the central processing of painful mechanical stimulation to muscle and bone by measuring blood oxygen level dependent signal changes using functional magnetic resonance imaging (fMRI). METHODS: Twelve healthy volunteers were enrolled. Mechanical pressure on muscle and bone were applied at the right lower leg by an algometer. Intensities were adjusted to cause weak and strong pain sensation at either target site in preliminary testing. Brain activation in response to mechanical nociceptive stimulation targeting muscle and bone were measured by fMRI and analyzed. RESULTS: Painful mechanical stimulation targeting muscle and bone activated the common areas including bilateral insula, anterior cingulate cortex, posterior cingulate cortex, secondary somatosensory cortex (S2), inferior parietal lobe, and basal ganglia. The contralateral S2 was more activated by strong stimulation than by weak stimulation. Some areas in the basal ganglia (bilateral putamen and caudate nucleus) were more activated by muscle stimulation than by bone stimulation. CONCLUSIONS: The putamen and caudate nucleus may have a more significant role in brain processing of muscle pain compared with bone pain. PMID- 21633874 TI - Modulation of C-reactive protein and tumour necrosis factor-alpha in collagen induced arthritis in Dark Agouti rats: impact of collagen concentration on severity of arthritis. AB - OBJECTIVES: The study investigated the effect of collagen-induced arthritis in Dark Agouti (DA) rats on the level of C-reactive protein and inflammatory cytokine tumour necrosis factor-alpha (TNF-alpha). SUBJECTS: Female Dark Agouti (DA) rats. METHODS: Three different dosages of (2 mg/kg of body weight, 3 mg/kg of body weight and 4 mg/kg of body weight) collagen and complete Freund's adjuvant suspension were tested. After 45 days, serum C-reactive protein, TNF alpha, superoxide dismutase and total glutathione assays were done. Radiographic and histopathological changes in the joints were compared. RESULTS: All three groups showed signs of arthritic changes, confirmed by histopathological and radiographic changes. Severe arthritic changes were seen in the rats injected with 4 mg/kg of body weight of collagen. There was a significant increase in C reactive protein, TNF-alpha, super oxide dismutase and total glutathione levels in the plasma in arthritis rats and the changes were more significant with 4 mg/kg of collagen. CONCLUSION: These results demonstrated that the optimal dose to inject to experimental animals in order to get server arthritic changes was 4 mg/kg of collagen with complete Freund's adjuvant suspension. Severe arthritis changes induced significant elevation in plasma C-reactive protein and TNF-alpha levels. PMID- 21633875 TI - Multiple triphenylphosphonium cations as a platform for the delivery of a pro apoptotic peptide. AB - PURPOSE: Triphenyl phosphonium cations (TPPs) are delocalized lipophilic cations that accumulate in the mitochondria of cells. We have explore the effect of increasing the number of TPPs on delivery of a cell-impermeable pro-apoptotic peptide to intact cells. METHODS: The pro-apoptotic peptide D-(KLAKLAK)(2) (KLA) was extended with 0-3 L-Lysines modified at their epsilon-amine with TPP. Peptides were studied in HeLa cells to determine their cytotoxic activity and cellular uptake. RESULTS: In HeLa cells, the increased cytotoxicity correlates with the number of TPPs; the peptide with 3 TPP molecules (3-KLA) exerts the highest cytotoxic activity. This FITC-labeled peptide is found to accumulate in intact HeLa cells, whereas peptides with 0-2 TPPs are not detected at the same peptide concentration. Mitochondria-dependent apoptosis of HeLa cells in the presence of 3-KLA was followed by propidium iodide, Annexin-V and DiOC fluorescence by FACS. CONCLUSION: A facile synthetic methodology has been presented for the delivery of a biologically active peptide into mitochondria of intact cells by attaching multiple TPP moieties to the peptide. This approach was shown to dramatically increase biological activity of the peptide as a pro apoptotic agent. PMID- 21633876 TI - Structure-skin permeability relationship of dendrimers. AB - PURPOSE: To investigate skin penetration of poly (amidoamine) (PAMAM) dendrimers as a function of surface charge and molecular weight in presence and absence of iontophoresis. METHODS: Dendrimers were labeled with fluoroisothiocynate (FITC); skin penetration of dendrimers was studied using excised porcine skin in-vitro. Skin penetration of FITC-labeled dendrimers was quantified using confocal laser scanning microscope (CLSM). G2-G6 NH(2), G3.5-COOH and G4-OH dendrimers were used. RESULTS: Cationic dendrimers showed higher skin penetration than neutral and anionic dendrimers. Skin penetration of cationic dendrimer increased linearly with increase in treatment time. Iontophoresis enhanced skin penetration of cationic and neutral dendrimers. Increase in current strength and current duration increased skin transport of dendrimers. Passive and iontophoretic skin penetration of cationic dendrimers was inversely related to their molecular weight. Dendrimer penetrated the skin through intercellular lipids and hair follicles. With iontophoresis, dendrimer was also found in localized skin regions. CONCLUSIONS: The study demonstrates that the physicochemical properties of dendrimers influence their skin transport. Findings can be used to design dendrimer-based nanocarriers for drug delivery to skin. PMID- 21633877 TI - Social relationships as a major determinant in the valuation of health states. AB - PURPOSE: To empirically determine the impact of the capacity to sustain social relationships on valuing health states. METHODS: 68 clinical experts conducted a health state valuation exercise in five sites using pairwise comparison, ranking, and person trade-off as elicitation methods. 23,840 pairwise comparisons of a total of 379 health states were analyzed by conditional logistic regression. RESULTS: Social relationships had a clear monotonic association with perceived disability: the more limited the capacity to sustain social relationships, the more disabling the resulting health state valuations. The highest level of limitations with respect to social relationships was associated with slightly lower impact on health state valuations compared to the highest level of limitations in physical functioning. CONCLUSIONS: Social relationships showed an independent contribution to health state valuations and should be included in health state measures. PMID- 21633878 TI - Differences in quality of life of hemodialysis patients between dialysis centers. AB - PURPOSE: Hemodialysis patients undergo frequent and long visits to the clinic to receive adequate dialysis treatment, medical guidance, and support. This may affect health-related quality of life (HRQOL). Although HRQOL is a very important management aspect in hemodialysis patients, there is a paucity of information on the differences in HRQOL between centers. We set out to assess the differences in HRQOL of hemodialysis patients between dialysis centers and explore which modifiable center characteristics could explain possible differences. METHODS: This cross-sectional study evaluated 570 hemodialysis patients from 24 Dutch dialysis centers. HRQOL was measured with the Kidney Disease Quality Of Life Short Form (KDQOL-SF). RESULTS: After adjustment for differences in case-mix, three HRQOL domains differed between dialysis centers: the physical composite score (PCS, P = 0.01), quality of social interaction (P = 0.04), and dialysis staff encouragement (P = 0.001). These center differences had a range of 11-21 points on a scale of 0-100, depending on the domain. Two center characteristics showed a clinical relevant relation with patients' HRQOL: dieticians' fulltime equivalent and the type of dialysis center. CONCLUSION: This study showed that clinical relevant differences exist between dialysis centers in multiple HRQOL domains. This is especially remarkable as hemodialysis is a highly standardized therapy. PMID- 21633879 TI - Quality of life in pregnancy and post-partum: a study in diabetic patients. AB - PURPOSE: We evaluated quality of life in pregnant women with diabetes followed up at Italian diabetes clinics. METHODS: A total of 245 pregnant women (30 type 1 diabetes mellitus (T1DM), 176 gestational diabetes (GDM) and 39 controls) were asked to fill in a questionnaire including the SF-36 Health Survey and the Center for Epidemiological Studies-Depression (CES-D) Scale in third trimester of pregnancy and after delivery. GDM and T1DM also completed two diabetes-specific questionnaires (Diabetes-related stress and Diabetes health distress). Quality of life scores were compared between the groups with the Mann-Whitney U-test, mean changes in scores (after delivery to 3rd trimester) were compared between groups by ANCOVA. RESULTS: Regarding the SF-36 scores in the third trimester of pregnancy, T1DM and GDM women had a better Standardised Physical Component score than controls (P < 0.0001, P = 0.009, respectively). GDM and T1DM pregnant women scored significantly lower for general health perception than controls (P = 0.009 and P = 0.001, respectively). T1DM patients had lower Standardised Mental Component scores than controls (P = 0.03). Compared with the third trimester of pregnancy, the severity of depressive symptoms increased significantly after delivery in both diabetic groups, but not in controls (P < 0.0001). Scores improved in all SF-36 areas in healthy and GDM women, while they all became worse in the T1DM group. CONCLUSIONS: Pregnancy is associated with a perception of poor general health in women with both T1DM and GDM. After delivery, significantly worse depressive symptoms were documented in both groups, while a generally worse physical and psychological well-being was only identified in women with T1DM. These findings have important implications for pregnancy follow-up. PMID- 21633880 TI - Determinants of social participation of visually impaired older adults. AB - PURPOSE: To assess determinants of social participation among visually impaired older adults. METHODS: This cross-sectional study included visually impaired persons (>=55 years; n = 173) who were referred to a low-vision rehabilitation center. Determinants (i.e., sociodemographic, physical, social and psychological factors, and personal values) of participation were identified in four domains of participation: (1) domestic life; (2) interpersonal interactions and relationships; (3) major life areas; and (4) community, social, and civic life. Study participants completed telephone interviews. RESULTS: Age, physical fitness, and helplessness were determinants of participation in domestic life. Social network size was associated with participation in major life areas. The personal value attached to participation (i.e., perceived importance) was a determinant of participation in interpersonal interactions and relationships, major life areas, and community, social and civic life. Vision-related characteristics (i.e., self-perceived vision and degree of visual impairment) were not associated with participation. CONCLUSIONS: Across the participation domains, perceived importance is a major determinant of social participation among visually impaired older adults. Physical health along with social and psychological status, also affect participation. Knowing how participation is determined can be used to develop rehabilitation interventions to enhance participation of visually impaired older adults. PMID- 21633881 TI - Fatal cardiac perforation after percutaneous treatment in iliac artery occlusion. PMID- 21633882 TI - Microwave ablation with percutaneous approach for the treatment of pancreatic adenocarcinoma. PMID- 21633883 TI - Angiography in the isolated perfused kidney: radiological evaluation of vascular protection in tissue ablation by nonthermal irreversible electroporation. AB - PURPOSE: The nonthermal irreversible electroporation (NTIRE) is a novel nonthermal tissue ablation technique by local application of high-voltage current within microseconds leading to a delayed apoptosis. The purpose of this experimental study was the first angiographic evaluation of the acute damage of renal vascular structure in NTIRE. METHODS: Results of conventional dynamic digital substraction angiography (DSA) and visualization of the terminal vascular bed of renal parenchyma by high-resolution X-ray in mammography technique were evaluated before, during, and after NTIRE of three isolated perfused porcine ex vivo kidneys. RESULTS: In the dedicated investigation, no acute vascular destruction of the renal parenchyma and no dysfunction of the kidney perfusion model were observed during or after NTIRE. Conspicuous were concentric wave-like fluctuations of the DSA contrast agent simultaneous to the NTIRE pulses resulting from NTIRE pulse shock wave. CONCLUSION: The NTIRE offers an ablation method with no acute collateral vascular damage in angiographic evaluation. PMID- 21633885 TI - Tracking miRNA precursor metabolic products and processing sites through completely analyzing high-throughput sequencing data. AB - The small non-coding important regulatory molecules, microRNAs (miRNAs), have been widely and deeply studied especially combining high-throughput sequencing technologies. Here, we attempted to track detailed miRNA precursor metabolic products and gain further insight into pre-miRNA processing by completely analyzing high-throughput sequencing data. Highly expressed miRNA precursors could be entirely covered by various short RNAs and small RNA fragments with a hierarchical distribution. miRNAs and some miRNA* regions were detected quite abundant short RNAs as expected, while other regions of precursors were found shorter RNAs or small fragments with fewer sequence counts. Furthermore, we developed a method to analyze relative expression levels of special RNA classes according to divergence of 5' and 3' ends, respectively. Generally, there were several quite abundant RNA classes from a given miRNA locus, which suggested dominant cleavage sites of Drosha and Dicer during pre-miRNA processing. Compared with 3' end, dominant cleavage site in 5' end always focused on a specific position, which ensured conservation of the identity of miRNA (5'-seed sequence, nucleotides 2-8). Overall, a comprehensive analysis of sequencing data can be used to track pre-miRNA metabolic products and mechanism of pre-miRNA processing and metabolism. PMID- 21633886 TI - Genetic variability of the coding region for the prion protein gene (PRNP) in gayal (Bos frontalis). AB - The gayal (Bos frontalis) is a rare semi-wild bovid species in which bovine spongiform encephalopathy (BSE) has not been reported. Polymorphisms of the prion protein gene (PRNP) have been correlated significantly with resistance to BSE. In this study, the coding region of PRNP was cloned and characterized in samples from 125 gayal. A total of ten single nucleotide polymorphisms (SNPs), including six silent mutations (C60T, G75A, A108T, G126A, C357T and C678T) and four mis sense mutations (C8A, G145A, G461A and C756G), corresponding to amino acids T3K, G49S9, N154S and I252M were identified, revealing high genetic diversity. Three novel SNPs including C60T, G145A and C756G, which have not been reported previously in bovid species, were retrieved. There also was one insertion deletion (187Del24) at the N-terminal octapeptide repeat region. Alignment of nucleotide and amino acid sequences showed a high degree of similarity with other bovid species. Using phylogenetic analyses it was revealed that gayal has a close genetic relationship with Zebu cattle. In short, preliminary information is provided about genotypes of the PRNP in gayal. This could assist with the study of the pathogenesis of transmissible spongiform encephalopathies and cross species transmission as well as a molecular breeding project for gayal in China. PMID- 21633887 TI - Use of siRNA in knocking down of dopamine receptors, a possible therapeutic option in neuropsychiatric disorders. AB - Heightened dopaminergic activity has been shown to be implicated in some major neuropsychiatric disorders such as schizophrenia. Use of dopaminergic antagonists was limited by some serious side effects related to unspecific blocking of dopamine receptors. Thus a target specific dopamine receptor gene silencing method such as using small interfering RNA (siRNA) might be useful. In this study recombinant plasmids expressing siRNA against dopamine receptors (D1-D5DRs) were produced, and their efficiency in knocking down of receptors in were assessed in rat neuroblastoma cell line (B65), using Real-time PCR method. Furthermore, D2DR siRNA expressing plasmid was injected into the rat nucleus accumbens bilaterally to investigate whether it can prevent the hyperactivity induced by apomorphine. Locomotion was measured in 10 min intervals, 50 min before and 60 min after apomorphine injection (0.5 mg/kg, S.C). Our results indicated that the mRNA level of dopamine receptors were reduced between 25 and 75% in B65 cells treated with the plasmids in vitro. In behavioral tests, locomotion was lower at least in the second 10 min after apomorphine injection in rats treated with plasmid expressing D2DR siRNA compare to control group [F (4,24) = 2.77, (P < 0.05)]. The spontaneous activity of treated rats was normal. In conclusion, dopamine receptors can be downregulated by use of siRNA expressing plasmids in nucleus accumbens. Although our work may have some possible clinical applications; the potentially therapeutic application of siRNA in knocking down of dopamine receptors needs further studies. PMID- 21633888 TI - Selection of reference genes for quantitative real-time RT-PCR analysis in citrus. AB - Quantitative real-time reverse transcription polymerase chain reaction (qPCR) has become the preferred method for studying low-abundant mRNA expression. Appropriate application of qPCR in such studies requires the use of reference gene(s) as an internal control in order to normalize the mRNA levels between different samples for an exact comparison of gene expression levels. Expression of the reference gene should be independent from development stage, cell/tissue types, treatments and environmental conditions. Recognizing the importance of reference gene(s) in normalization of qPCR data, various reference genes have been evaluated for stable expression under specific conditions in various organisms. In plants, only a few of them have been investigated, and very few reports about such reference genes in citrus. In the present study, seven candidate reference genes (18SrRNA, ACTB, rpII, UBQI, UBQ10, GAPDH and TUB) were tested, and three of them (18SrRNA, ACTB and rpII) proved to be the most stable ones among six leaf samples of different citrus genotypes. The three candidate reference genes were further analyzed for their stability of expression in five different tissues, and the results indicated that they were not completely stable. It is commonly accepted that gene expression studies should be normalized using more than one reference gene. Based on our results, we propose the use of the mean result rendered by18SrRNA, ACTB and rpII as reference genes to normalize mRNA levels in qPCR analysis of diverse cultivars and tissues of citrus. These results may provide a guideline for future works on gene expression in citrus by using qPCR. PMID- 21633889 TI - Calcium ameliorates obesity induced by high-fat diet and its potential correlation with p38 MAPK pathway. AB - To investigate whether and on which pathway dietary calcium influence the obesity induced by high-fat diet, thirty male Kunming mice were fed in six groups for 4 weeks and mouse preadipocytes were divided into eight groups for different treatment. Body weight gain was measured each week. Calcium in serum and tissues, intracellular free Ca(2+) concentration ([Ca(2+)]i), blood fat and intracellular lipid content were also measured. The expression of Lipid metabolism-related genes were measured by q RT-PCR. Compared with control group, body weight gain (P < 0.05) and fat pad weight (P < 0.01) in Low calcium group decreased. Triglycerides (TG) and total Cholesterol (TC) level decreased (P < 0.01), while HDL-Cholesterol (HDL) level increased (P < 0.01). And calcium supply increased calcium content in blood serum and tissues. In tissues, adipogenesis and vitamin D receptor (VDR) genes expression decreased but lipoclasis genes expression increased. These anti-obesity effects were more obvious when supplying with 2.8% calcium, but the effects were reduced while supplying Nifedipine at the same time. The results in preadipocytes indicated that calcium-treated can reduce intracellular lipid content, along with adipogenesis and lipoclasis genes expression decrease, promoted the expression levels of p38 MAPK pathway upstream gene MKK6 (P < 0.01) and downstream gene MAPKAPK2 (P < 0.01). Treated with SB203580 could increase adipogenesis genes expression, decrease lipoclasis genes expression and ([Ca(2+)]i) (P < 0.01). These results implied that dietary calcium had remarkable effect on anti-obesity effect and p38 MAPK pathway potentially participated in calcium-mediated lipid accumulation and lipolysis in mouse preadipocytes. PMID- 21633890 TI - The apoptosis inhibitor survivin prevents insect midgut from cell death during postembryonic development. AB - The programmed cell death (PCD) is important in maintaining the cell number homoeostasis of tissues and organs in metazoan. This process is regulated by the inhibitors of apoptosis proteins (IAPs). The function and mechanism of IAPs have been well studied in animal embryonic development and human cancers. However, the roles and hormonal regulation of IAPs in the postembryonic development are not well understood. We report that an IAP survivin (Ha-survivin) played roles in the postembryonic development of the midgut in a lepidopteran insect Helicoverpa armigera. Ha-survivin was transcribed not only in the embryo, but also in the haemocytes, fat body and midgut during larval molting or pupal adulting. The transcription of Ha-survivin was upregulated by the steroid hormone 20 hydroxyecdysone (20E). Ha-survivin was located in the embryonic cells around the periplasm of the eggs during embryonic development. It was also located in the epithelium cells of the midgut in the fifth molting larvae and later pupae. Knockdown of Ha-survivin by RNAi in the epidermal cell line caused cell apoptosis. These results indicated that Ha-survivin played roles not only in the embryonic development, but also in the postembryonic tissue development by preventing cell death. PMID- 21633891 TI - Functional analysis of the HS185 regulatory element in the rice HSP70 promoter. AB - A series of HSP70 promoter deletion constructs was established. Analysis of beta glucuronidase activities from the promoter deletion constructs in transient expression assays identified a cis-element, located from -493 to -308 bp upstream of the ATG start site. This element was designated as HS185 and has a crucial role in HSP70 promoter activity. HS185 has some characteristics of a miniature inverted-repeat transposable element (MITE), such as terminal inverted repeats (TIRs) (GGTCCCACA) and a putative target site duplication. There are 362 copies of homologous sequences of HS185 in the rice genome, which are preferentially distributed to non-coding regions. Based on these sequence features, we propose that HS185 is an uncharacterized rice MITE, possibly derived from the rice transposon Mutator-like element VIII family. Further transient expression assays showed that HS185 inhibited the enhancer activity of the cauliflower mosaic virus 35S promoter. These results demonstrate that not only is HS185 necessary for HSP70 promoter activity, but it also has a functional role as an insulator. This study explored new regulatory functions of non-coding repeat sequences in rice. PMID- 21633892 TI - A meta-analysis on the association between three promoter variants of TNF-alpha and Crohn's disease. AB - Tumor necrosis factor-alpha (TNF-alpha) has been regarded as a candidate gene for Crohn's disease (CD) based on its inflammatory function in immune reaction and the clinical effectiveness of anti-TNF-alpha therapy. However, studies to date have reported inconsistent findings for the association between TNF-alpha and CD. The PubMed, EMBASE, and Medline databases were systematically reviewed from all English language publications up to April, 2011. A total of twenty-nine studies concerning the association between CD and the TNF-alpha promoter polymorphisms of -308G/A, -857C/T and -238G/A were identified, among of them only twenty-three studies match the inclusion criteria (including 3,843 cases and 6,260 controls) and were selected for the statistical test. We found that neither the G allele of -308G/A (OR 1.02, 95% CI 0.87-1.19, P = 0.84), C allele of -857C/T (OR 0.97, 95% CI 0.86-1.09, P = 0.57) and G allele of -238G/A (OR 0.91, 95% CI 0.70-1.18, P = 0.48), and nor their GG (OR 1.05, 95% CI 0.88-1.25, P = 0.59), CC (OR 0.98, 95% CI 0.86-1.12, P = 0.76) and GG (OR 0.92, 95% CI 0.70-1.21, P = 0.55) genotypes were associated with CD susceptibility, respectively. Our meta-analysis demonstrates that three promoter polymorphisms of TNF-alpha above may not confer susceptibility to CD. PMID- 21633893 TI - MUC1 gene polymorphism in three Nelore lines selected for growth and its association with growth and carcass traits. AB - The objective of this study was to describe the VNTR polymorphism of the mucin 1 gene (MUC1) in three Nelore lines selected for yearling weight to determine whether allele and genotype frequencies of this polymorphism were affected by selection for growth. In addition, the effects of the polymorphism on growth and carcass traits were evaluated. Birth, weaning and yearling weights, rump height, Longissimus muscle area, backfat thickness, and rump fat thickness, were analyzed. A total of 295 Nelore heifers from the Beef Cattle Research Center, Instituto de Zootecnia de Sertaozinho, were used, including 41 of the control line, 102 of the selection line and 152 of the traditional. The selection and traditional lines comprise animals selected for higher yearling weight, whereas control line animals are selected for yearling weight close to the average. Five alleles were identified, with allele 1 being the most frequent in the three lines, especially in the lines selected for higher means for yearling weight. Heterozygosity was significantly higher in the control line. Association analyses showed significant effects of allele 1 on birth weight and weaning weight while the allele 3 exert significant effects on yearling weight and back fat thickness. Despite these findings, application of this marker to marker-assisted selection requires more consistent results based on the genotyping of a larger number of animals in order to increase the accuracy of the statistical analyses. PMID- 21633894 TI - Las Alpujarras region (South East Spain) HLA genes study: evidence of a probable success of 17th century repopulation from North Spain. AB - Conquest of Granada Muslim Kingdom (1492 AD) finished with Muslim occupation; they were mostly North African Berbers who had reached Iberia by 711 AD. A politics of Iberian Christianization followed after this date: Jewish were expelled in 1492 and Moriscos (Spaniards practicing Muslim religion or speaking Arab) were expelled from all Spanish territory on 1609 AD. Las Alpujarras is a southern Spain mountainous secluded region, which underwent a repopulation from North Spain and a specific Muslim (Moriscos)-Christian war took place according to historical records. Both Las Alpujarras repopulation by northern Iberians and Moriscos expulsion success have been debated and are regarded as non-clarified episodes. In this study, we have addressed the question whether the repopulation succeeded by determining HLA genes of present day Las Alpujarras inhabitants and compared with those of other Mediterranean populations HLA frequencies and genealogies. HLA frequencies show ambiguous results because of extant HLA similar gene frequencies there exist in North Africa and Spain. This is reflected by the finding of North and South western Mediterraneans close relatedness of HLA dendrograms and correspondence analyses. However, the genealogical study of extended HLA haplotypes particularly Alpujarran high frequency of HLA-A29-B44 DRB1*0701-DQA1*02-DQB1*02 (not found in Algerians but frequent in North and Central Spain) and Alpujarran low frequency extended haplotype HLA-A3-B7 DRB1*1501-DQA1*0102-DQB1*0602 (frequent in North Europe) reveals that a significant HLA gene flow from North Spain is observed in present day Alpujarrans: both haplotypes are characteristic of North Spain and North Europe, respectively. This may indicate that enforced Alpujarran repopulation from North Spain may have been a success, which was started by Spanish King Philip II in 1571 AD. PMID- 21633895 TI - MicroRNA regulated defense responses in Triticum aestivum L. during Puccinia graminis f.sp. tritici infection. AB - Plants have evolved diverse mechanism to recognize pathogen attack and triggers defense responses. These defense responses alter host cellular function regulated by endogenous, small, non-coding miRNAs. To understand the mechanism of miRNAs regulated cellular functions during stem rust infection in wheat, we investigated eight different miRNAs viz. miR159, miR164, miR167, miR171, miR444, miR408, miR1129 and miR1138, involved in three different independent cellular defense response to infection. The investigation reveals that at the initiation of disease, accumulation of miRNAs might be playing a key role in hypersensitive response (HR) from host, which diminishes at the maturation stage. This suggests a possible host-fungal synergistic relation leading to susceptibility. Differential expression of these miRNAs in presence and absence of R gene provides a probable explanation of miRNA regulated R gene mediated independent pathways. PMID- 21633896 TI - Selection of housekeeping genes for normalization of RT-PCR in hypoxic neural stem cells of rat in vitro. AB - Gene expression analysis under various conditions using real-time reverse transcription polymerase chain reaction (RT-PCR) needs reliable control genes. Housekeeping genes are commonly used as the control. However, no validated housekeeping gene is available for study of hypoxic neural stem cell culture. To choose appropriate internal control genes, the expression of eight commonly used housekeeping genes was examined in rat neural stem cell model to find one or more stably expressed genes under hypoxic/ischemic conditions. Two genes, HPRT and RPL13A were identified as the most confidential housekeeping genes in this research by geNorm and NormFinder softwares. As a groundwork, the most stable housekeeping genes for neural stem cells under hypoxic/ischemic conditions are initially investigated and validated in this experiment, which might provide a better understanding for the gene expression study in ischemic and necrotic neural stem cell cultures or in ischemic diseases of the central nervous system (CNS). PMID- 21633897 TI - Imprinting analysis of porcine MAGEL2 gene in two fetal stages and association analysis with carcass traits. AB - Imprinted genes play an essential role in the regulation of fetal growth, development and function of the placenta, however only a limited number of imprinted genes have been studied in swine. In this study, we cloned and characterized porcine MAGEL2 (melanoma antigen-like gene 2), and also identified its imprinting status during porcine fetal development. The complete open reading frame (ORF) encoding 1,193 amino acids was isolated and two single nucleotide polymorphisms (SNPs) (g.2592A>C and g.3277T>C) in the coding region were identified. The reciprocal Yorkshire*Meishan F1 hybrid model and the RT-PCR/RFLP method were used to detect the imprinting status of porcine MAGEL2 gene at two developmental stages of day 30 and 65 of gestation. Imprinting analysis showed that porcine MAGEL2 was paternally expressed in day 65 fetal tissues, including heart, liver, spleen, lung, kidney, stomach, small intestine, skeletal muscle, brain and placenta. Interestingly, we observed an imprinting variance of MAGEL2 gene in 30 dpc fetuses produced by the cross of Yorkshire boar*Meishan sow, in which seven heterozygous fetuses were monoallelically expressed from the paternal allele but two were biallelically expressed from both the paternal and maternal alleles. Association analysis in a Yorkshire*Meishan F2 resource population showed that the mutation of g.2592A>C was significantly associated with dressed carcass percentage (P<0.05) and buttock fat thickness (P<0.05). Our results suggest that MAGEL2, as a novel imprinted gene in pig, might be a candidate gene affecting carcass traits and could provide important information for the functional study of imprinted genes during porcine development. PMID- 21633898 TI - Molecular characterization and transcriptional analysis of the olive flounder (Paralichthys olivaceus) YGHL1 gene in response to hypoxia and infection. AB - The hypoxia-inducible gene 1, YGHL1 from olive flounder (Paralichthys olivaceus) (fYGHL1) was cloned, and its structural organization and expression profiles were determined. A 1,400 kb full-length cDNA encoding a predicted polypeptide of 91 amino acids was sequenced. The fYGHL1 gene comprises three introns, four exons, and several transcriptional elements upstream of the transcriptional start site. The mRNA transcript is expressed in almost all tissues, with high expression in the intestine and brain of normal-conditioned fish, and is expressed constitutively in early developmental stages after hatching. The mRNA expression of fYGHL1 is highly regulated by hypoxia and E. tarda infection. The expression of fYGHL1 mRNA was down regulated in the gill, spleen, intestine, and stomach of flounder under hypoxic conditions, whereas the expression level was increased in flounder embryonic cells treated with the hypoxia-mimic CoCl(2) (a HIF-1 inducer). Pathogen challenge induced fYGHL1 expression in the spleen of juvenile fish. Taken together, these results suggest that fYGHL1 is a hypoxia-related gene with potential roles in the hypoxia response mechanism as well as in defense, immune responses, growth, and regulation of reproduction. PMID- 21633899 TI - Comparison of adipocyte-specific gene expression from WNIN/Ob mutant obese rats, lean control, and parental control. AB - Adipose tissue development is a highly regulated phenomenon orchestrated by several check points (recruitment of mesenchymal stem cells and their lineage commitment) to form mature adipocytes. Once committed to obesity, expansion of adipose tissue occurs either by hypertrophy or hyperplasia or by both resulting in an altered physiological status. This precipitates as inflammatory responses, leading to endoplasmic reticulum and oxidative stress altering the gene expression of adipose tissue in a depot-specific manner. However, such studies reporting a phased gene expression profile in conditions of rodent obesity are not reported so far. WNIN/Ob mutant obese rat, developed at our institute is an excellent model to study the pathophysiological changes underlying obesity. Here, we report the gene expression profile of this mutant rat (obese and lean), compared with the parental control, with reference to markers of embryonic stem cells, adipogenesis, inflammation, and senescence in both subcutaneous (SCAT) and retroperitoneal (RPAT) adipose depots representing abdominal fat. We demonstrate an upregulation of genes such as Sox-2, Pref-1, PPARgamma2, LPL, IRS-1, GLUT-4, IL-6, TNFalpha, and telomerase in SCAT and RPAT depots of the obese rat compared to its lean counterpart indicating no difference in fat depots at different locations. This is suggestive of a similar phenotypic expression of mutant gene. Data form the phased gene expression changes of adipogenesis (embryonic/adipogenic/inflammatory) in the present obese rat model system advocate for inflammatory mediated response(s) associated with obesity-a condition often seen in humans. PMID- 21633900 TI - Eukaryotic translation elongation factor 1 delta inhibits the ubiquitin ligase activity of SIAH-1. AB - SIAH-1, an E3 ubiquitin ligase, plays an important role in regulating cell cycle, tumorigenesis and several neurodegenerative diseases. In this study, we found a novel SIAH-1-interacting protein, EEF1D (Eukaryotic translation elongation factor 1 delta). The interaction was confirmed in vitro and in vivo, and both proteins were co-localized in the cytoplasm. The Cys-rich domain of SIAH-1 was essential for its interaction with EEF1D. Overexpressing SIAH-1 had no effect on the protein level of EEF1D, implying that EFF1D is not the substrate of SIAH-1. In contrast, the protein level of SIAH-1 increased significantly in the cells overexpressing EEF1D. Increased amount of SIAH-1 was caused by the EEF1D-mediated inhibition of auto-ubiquitination and degradation of SIAH-1. Furthermore, EEF1D was able to inhibit the degradation of HPH2, a known substrate of SIAH-1. Taken together, our data suggest EFF1D functions as a novel negative regulator of SIAH 1. PMID- 21633901 TI - Attenuation of the cardiac inflammatory changes and lipid anomalies by (-) epigallocatechin-gallate in cigarette smoke-exposed rats. AB - Cigarette smoking is a major risk factor for cardiovascular diseases and exerts negative effects on the lipid profile. This study was aimed to evaluate the preventive role of (-)-epigallocatechin-gallate (EGCG) on lipid metabolism and cardiac inflammatory changes in cigarette smoke (CS) induced myocardial dysfunction. Adult male albino rats were exposed to side stream CS for a period of 12 weeks and simultaneously administered with EGCG (20 mg/kg b.w./day, p.o.). Exposure to CS showed significant increased (P < 0.05) activities of cardiac injury markers such as, creatine kinase-MB (CKMB) and lactate dehydrogenase (LDH) in serum and subsequent decrease in these enzyme activities in heart. A significant increase (P < 0.05) in serum total cholesterol, fatty acids, phospholipids, and triglycerides were observed in CS exposed rats, along with elevated low-density lipoprotein (LDL) and very low-density lipoprotein (VLDL) cholesterol and decreased high density lipoprotein (HDL) cholesterol. In myocardium, total cholesterol, fatty acids and triglycerides were increased, whereas the phospholipids were found to be decreased. Cardiac lecithin: cholesterol acyl trasferase (LCAT), lipoprotein lipase (LPL), and plasma LCAT activities were significantly decreased (P < 0.05) on CS exposure. Supplementation of EGCG reverted the cardiac injury markers, abnormalities of lipid profile, and lipid-metabolizing enzymes in serum and myocardium. Western blot analysis showed a significant increase in protein expression levels of nuclear factor kappa-B (NF-kappaB), cyclooxygenase-2 (COX-2), tumor necrosis factor-alpha (TNF-alpha), and inducible nitric oxide synthase (iNOS) in heart of CS exposed rats. EGCG-treated rats showed a significant decrease in the expression of inflammatory markers. Our data suggest that chronic CS causes lipidemic anomalies and cardiac inflammatory aberrations which may promote cardiac dysfunction and that the antioxidant EGCG exerts a cardio protective effect via reduction of oxidative stress. PMID- 21633902 TI - Anhidrosis associated with hypothalamic lesions related to anti-aquaporin 4 autoantibody. PMID- 21633903 TI - Subclinical nigrostriatal dopaminergic denervation in the cerebellar subtype of multiple system atrophy (MSA-C). AB - Nigrostriatal involvement is considered an additional feature in the new consensus criteria for the diagnosis of the cerebellar variant of multiple system atrophy (MSA-C). However, so far, only a few studies, which include a relative small number of patients, give support to this criterion. Our objective was to assess nigrostriatal dopaminergic innervation in patients with MSA-C without parkinsonism by use of dopamine transporter single photon emission computed tomography (DAT SPECT). Thirteen patients that fulfilled criteria for possible or probable MSA-C and presented no parkinsonian signs, and 12 age-matched healthy controls underwent ((123)I-2-beta-carbomethoxy-3beta-(4-iodophenyl)-N-(3 fluoropropyl) nortropane ([(123)I]FP-CIT) SPECT. Patients were also evaluated through the Unified Multiple System Atrophy Rating Scale (UMSARS) and brain magnetic resonance imaging (MRI). The mean duration of the cerebellar syndrome was 3.8 +/- 1.7 years. DAT SPECT showed a significant decrease of striatal [(123)I]FP-CIT uptake ratios in patients (p < 0.001). Radiotracer uptake reduction was 21% in the entire striatum, 19% in putamen, and 24% in caudate nuclei. Striatal binding ratios were within the normal range in 3 patients. We did not find correlation between striatal uptake and disease duration, age of patients, UMSARS-II score, and pontine diameter. [(123)I]FP-CIT SPECT shows that most but not all MSA-C patients without parkinsonism have subclinical nigrostriatal dopaminergic denervation which is not related to disease duration, cerebellar dysfunction, or pontine atrophy. PMID- 21633904 TI - Congenital mirror movements: a clue to understanding bimanual motor control. AB - Mirror movements (MM) are involuntary movements of one side of the body that accompany and mirror intentional movements on the opposite side. Physiological MM can occur during normal childhood development, probably owing to corpus callosum immaturity. Pathological congenital MM may be clinically isolated or part of a complex congenital syndrome, including Kallmann syndrome, Klippel-Feil syndrome, and congenital hemiplegia. Congenital isolated MM are usually familial. Recently, heterozygous mutations of the DCC gene, with autosomal dominant inheritance, were shown to cause some cases of MM. The pathogenesis of congenital MM may involve (i) abnormal interhemispheric inhibition between the two motor cortices; (ii) functional alteration of motor planning and motor execution; and/or (iii) abnormal persistence of the ipsilateral corticospinal tract. Fundamental and clinical research is providing novel insights into the complex underlying molecular pathways, and recent experimental work has identified several mechanisms that may mediate the motor network dysfunction. In this review, we analyze clinical, genetic, neurophysiologic, and neuroimaging data on congenital MM, and discuss how this knowledge may improve our understanding of bimanual motor control. PMID- 21633905 TI - Quantification of adverse events associated with functional MRI scanning and with real-time fMRI-based training. AB - BACKGROUND: Although functional magnetic resonance imaging (fMRI) is in widespread research use, the safety of this approach has not been extensively quantitatively evaluated. Real-time fMRI (rtfMRI)-based training paradigms use fMRI neurofeedback and cognitive strategies to alter regional brain activation, and are currently being evaluated as a novel approach to treat neurological and psychiatric conditions. PURPOSE: The purpose of this study is to determine the incidence and severity of any adverse events that might be caused by changes in brain activation brought about through fMRI or through rtfMRI-based training paradigms. METHOD: Quantitative adverse event self-report data were obtained from 641 functional imaging scans in 114 chronic pain patients participating in a research clinical trial examining repeated fMRI scans and rtfMRI-based training. Participants recorded potential adverse events during non-scanning baseline, fMRI scanning, or rtfMRI-based training sessions. RESULTS: There were no significant increases in the number of reported adverse events following fMRI or rtfMRI scanning sessions compared to baseline non-scanning sessions in a chronic pain trial (N = 88). There were no reported adverse events of any kind for over 90% of sessions during the course of rtfMRI-based training. When adverse events were reported, they were almost exclusively mild or moderate in severity and similar to those observed in a non-scanning baseline session. There was no increase in adverse events reported by participants receiving feedback from any of four brain regions during repeated rtfMRI-based training scans compared to non-scanning baseline sessions. For chronic pain patients completing the rtfMRI-based training paradigm including up to a total of nine scan sessions (N = 69), neither the number nor severity of reported events increased during the fMRI or rtfMRI scanning portions of the paradigm. There were no significant increases in the number of reported adverse events in participants who withdrew from the study. CONCLUSION: Repeated fMRI scanning and rtfMRI training, consisting of repeated fMRI scanning in conjunction with cognitive strategies and real-time feedback from several regions of interest in multiple brain systems to control brain region activation, were not associated with an increase in adverse event number or severity. These results demonstrate the safety of repetitive fMRI scanning paradigms similar to those in use in many laboratories worldwide, as well as the safety rtfMRI-based training paradigms. PMID- 21633906 TI - Bortezomib stabilizes NOXA and triggers ROS-associated apoptosis in medulloblastoma. AB - We have previously demonstrated that bortezomib, a 26S proteasome inhibitor, effectively inhibits medulloblastoma growth in vivo in a genetically engineered Ptch1, p53 mouse model; however, bortezomib is also associated clinically with severe peripheral neuropathy, which would be disadvantageous for patients with central nervous system malignancy. The purpose of this study was to determine the mechanism of bortezomib efficacy in medulloblastoma in order to replicate more specifically the therapeutic advantage of targeting the ubiquitin-proteosome system. In our studies of upstream components of the ubiquitin-proteasome system, we identified the pro-apoptotic protein NOXA as a post-translationally modified target that is stabilized by bortezomib and induces caspase cleavage in the context of reactive oxidative stress induced cell death. These preclinical results may apply to the sizable fraction of Shh-driven human medulloblastoma and perhaps other medulloblastoma subtypes, independent of p53 status. PMID- 21633907 TI - Chloroplast beta chaperonins from A. thaliana function with endogenous cpn10 homologs in vitro. AB - The involvement of type I chaperonins in bacterial and organellar protein folding has been well-documented. In E. coli and mitochondria, these ubiquitous and highly conserved proteins form chaperonin oligomers of identical 60 kDa subunits (cpn60), while in chloroplasts, two distinct cpn60 alpha and beta subunit types co-exist together. The primary sequence of alpha and beta subunits is ~50% identical, similar to their respective homologies to the bacterial GroEL. Moreover, the A. thaliana genome contains two alpha and four beta genes. The functional significance of this variability in plant chaperonin proteins has not yet been elucidated. In order to gain insight into the functional variety of the chloroplast chaperonin family members, we reconstituted beta homo-oligomers from A. thaliana following their expression in bacteria and subjected them to a structure-function analysis. Our results show for the first time, that A. thaliana beta homo-oligomers can function in vitro with authentic chloroplast co chaperonins (ch-cpn10 and ch-cpn20). We also show that oligomers made up of different beta subunit types have unique properties and different preferences for co-chaperonin partners. We propose that chloroplasts may contain active beta homo oligomers in addition to hetero-oligomers, possibly reflecting a variety of cellular roles. PMID- 21633908 TI - Insulin X10 revisited: a super-mitogenic insulin analogue. AB - The molecular safety of insulin analogues has received a great deal of attention over the last year. In particular, attention has been directed to the mitogenic properties of insulin analogues as compared with human insulin. Understanding the mechanisms implicated in mediating mitogenic effects of insulin is therefore of particular interest. In this review we detail the story of the rapid-acting insulin analogue known as X10, which was the first insulin analogue in clinical development, but ended up being discontinued at an early clinical development stage following findings of mammary tumours in female Sprague-Dawley rats. The molecular characteristics of insulin X10, along with its interaction at both the IGF-1 receptor and the insulin receptor, have provided us with important insights into mechanisms implicated in metabolic and mitogenic signalling of insulin analogues. PMID- 21633909 TI - Characterisation of glyoxalase I in a streptozocin-induced mouse model of diabetes with painful and insensate neuropathy. AB - AIMS/HYPOTHESIS: Diabetic peripheral neuropathy (DN) is a common complication of diabetes; however, the mechanisms producing positive or negative symptoms are not well understood. The enzyme glyoxalase I (GLO1) detoxifies reactive dicarbonyls that form AGEs and may affect the way sensory neurons respond to heightened AGE levels in DN. We hypothesised that differential GLO1 levels in sensory neurons may lead to differences in AGE formation and modulate the phenotype of DN. METHODS: Inbred strains of mice were used to assess the variability of Glo1 expression by quantitative RT-PCR. Non-diabetic C57BL/6 mice were used to characterise the distribution of GLO1 in neural tissues by immunofluorescence. Behavioural assessments were conducted in diabetic A/J and C57BL/6 mice to determine mechanical sensitivity, and GLO1 abundance was determined by western blot. RESULTS: GLO1 immunoreactivity was found throughout the nervous system, but selectively in small, unmyelinated peptidergic dorsal root ganglia (DRG) neurons that are involved in pain transmission. GLO1 protein was present at various levels in DRG from different inbred mice strains. Diabetic A/J and C57BL/6 mice, two mouse strains with different levels of GLO1, displayed dramatically different behavioural responses to mechanical stimuli. Diabetic C57BL/6 mice also had a reduced abundance of GLO1 following diabetes induction. CONCLUSIONS/INTERPRETATION: These findings reveal that the abundance of GLO1 varies between different murine strains and within different sensory neuron populations. These differences could lead to different responses of sensory neurons to the toxic effects of hyperglycaemia and reactive dicarbonyls associated with diabetes. PMID- 21633910 TI - Intact internal dynamics of the neocortex in acutely paralyzed mice. AB - Animals collect sensory information through self-generated movements. Muscle movements drive active feedback of sensory information and determine large parts of the sensory inputs the animal receives; however, little is known about how this active feedback process modulates the ongoing dynamics of the brain. We made electrophysiological recordings from layer 2/3 neurons of the mouse neocortex and compared spontaneous cortical activity in local field potentials and intracellular potential fluctuations between normal and hypomyotonic conditions. We found that pancuronium-induced paralysis did not affect the electrophysiological properties of ongoing cortical activity and its perturbation evoked by visual and tactile stimuli. Thus, internal cortical dynamics are not much affected by active muscle movements, at least, in an acute phase. PMID- 21633911 TI - Six-transmembrane epithelial antigen of prostate4 (STEAP4) is a tumor necrosis factor alpha-induced protein that regulates IL-6, IL-8, and cell proliferation in synovium from patients with rheumatoid arthritis. AB - Human six-transmembrane epithelial antigen of prostate4 (STEAP4), an ortholog of mouse tumor necrosis factor-alpha-induced adipose-related protein (TIARP), plays a role in tumor necrosis factor (TNF)-dependent arthritis models. However, its role in rheumatoid arthritis (RA) is still obscure. This study explored such a role for STEAP4. The expressions of STEAP4, TNFalpha, and IL-6 were compared in synovia of RA and osteoarthritis patients. STEAP4 induction was examined in TNFalpha-stimulated fibroblast-like synoviocytes (FLS) in vitro. FLS (with/without TNFalpha stimulation) were also analyzed for IL-6 expression after STEAP4 knockdown, using siRNA or transfection with STEAP4-plasmid DNA. IL-8, cell proliferation, and apoptosis were also evaluated in STEAP4-overexpressing FLS. The expression of STEAP4 in joints correlated with TNFalpha expression, specifically in RA synovium. In the cultured FLS, STEAP4 protein expression was augmented by TNFalpha activation, and localized in endosomal/lysosomal compartments. STEAP4 downregulation by siRNA enhanced the expression of IL-6 mRNA, while STEAP4 overexpression suppressed IL-6 and IL-8 expression, inhibited cell proliferation, and induced apoptosis via caspase-3. The results indicated that human STEAP4 is regulated by TNFalpha in synovium, where it controls IL-6 secretion and proliferation of FLS, suggesting that STEAP4 might potentially suppress the pathogenesis of TNFalpha-induced arthritis such as RA. PMID- 21633912 TI - Efficacy of low-dose imatinib mesylate for cutaneous involvement in systemic sclerosis: a preliminary report of three cases. AB - In this pilot study, the effect of low-dose imatinib mesylate (100 mg/day) on cutaneous involvement in patients with systemic sclerosis (SSc) was analyzed. Three patients with SSc were treated with 100 mg/day of imatinib mesylate for 6 months because of pulmonary arterial hypertension refractory to conventional treatments, including beraprost, bosentan, sildenafil, and epoprostenol. Changes in cutaneous involvement were evaluated at 1, 3, and 6 months. During the treatment, the total skin score gradually improved in all of the patients. Contracture of phalanges was attenuated in two patients, one of whom also experienced the partial restoration of large-joint mobility. Nailfold bleeding, initially seen in two patients, was gradually attenuated and had completely disappeared at 6 months. In all patients, Raynaud's phenomenon was attenuated at around 3 months and had completely disappeared at 6 months. Although transient renal dysfunction was observed in one patient, none of the patients experienced common adverse effects of imatinib, such as edema, nausea, rash, and musculoskeletal pain. These clinical data indicate the tolerability and efficacy of low-dose imatinib in SSc, especially against cutaneous vascular involvement, including Raynaud's phenomenon and nailfold bleeding. PMID- 21633913 TI - Serum adipokine profiles in Kawasaki disease. AB - Adipokines are cytokines derived from adipose tissue. Recently it has been established that adipokines are closely linked to the pathophysiology of not only metabolic diseases, such as diabetes mellitus, obesity, and atherosclerosis, but also to inflammation and immune diseases. In this study we measured serum levels of adipokines in patients with acute Kawasaki disease to investigate the role of adipokines in the pathophysiology of Kawasaki disease. Serum resistin, high molecular-weight (HMW) adiponectin, leptin, and visfatin levels were measured by enzyme-linked immunosorbent assay in a total of 117 subjects: 56 patients with acute Kawasaki disease, 30 healthy children, and 31 patients with acute infectious diseases. Serum resistin levels in patients with Kawasaki disease were significantly higher than those of healthy children and patients with acute infectious diseases. In contrast, mean serum HMW adiponectin, leptin, and visfatin levels in patients with Kawasaki disease exhibited no statistically significant differences compared with those in healthy children and patients with infectious diseases. Serum resistin levels decreased significantly after administration of intravenous immune globulin. Serum resistin levels on admission were significantly higher in nonresponders compared with responders to intravenous immune globulin therapy. A multivariate model revealed that C reactive protein was a factor that was significantly related to elevated serum resistin level in patients with Kawasaki disease. In patients with Kawasaki disease, serum resistin levels were elevated, but decreased to nearly normal after intravenous administration of immune globulin. In contrast, serum HMW adiponectin, leptin, and visfatin levels showed no statistically significant changes. These findings suggest that resistin plays an important role, while other adipokines do not play a major role, in the pathogenesis of Kawasaki disease. PMID- 21633914 TI - Determination of L-tetrahydropalmatine in human plasma by HPLC and pharmacokinetics of its disintegrating tablets in healthy Chinese. AB - The objective of the study was to establish an HPLC method for the determination of L: -tetrahydropalmatine in human plasma, and to investigate the pharmacokinetics after oral administration of L: -tetrahydropalmatine disintegrating tablets in healthy Chinese. L: -tetrahydropalmatine in human plasma was separated on a Phenomenex luna C(18) column (250 mm * 4.6 mm, 5 MUm), eluted using methanol-water (75:25, v/v) as mobile phase, and detected by photodiode array detector at a wavelength of 281 nm. A single 60 mg of L: tetrahydropalmatine orally disintegrating tablets were orally given to 12 healthy male volunteers after fasting overnight. Before and after administration 4 mL of blood samples was collected at the scheduled time. The plasma concentration of L: -tetrahydropalmatine was determined by the established HPLC method after disposition and its pharmacokinetic parameters were analyzed and evaluated by both compartmental and noncompartmental models using Drug and Statistic (version 2.0). The disintegrating time and the sense of mouth were observed and recorded. The lowest limit of quantification (LLOQ) for L: -tetrahydropalmatine in plasma was 0.01 MUg mL(-1), and a linearity was obtained in the range of 0.01-1 MUg mL( 1) (r = 0.9998). The disposal procedure of L: -tetrahydropalmatine in human was fitted using the DAS program, following a double-compartment open model system (w = 1). L: -tetrahydropalmatine was absorbed quickly with t (1/2ka) of 0.5 +/- 0.054 h, distributed fast with t (1/2alpha) of 0.74 +/- 0.088 h, and eliminated slowly with t (1/2beta) of 11.42 +/- 2.43 h. L: -tetrahydropalmatine was distributed mainly in the periphery compartment with the V(1)/F of 133.30 +/- 30.78 L. L: -tetrahydropalmatine orally disintegrating tablets with good taste were disintegrated in the mouth within 16 s. The established HPLC method was sensitive, rapid, and suitable for both L: -tetrahydropalmatine pharmacokinetic studies and its content assay in traditional Chinese medicine (TCM). The procedure of L: -tetrahydropalmatine in human was fit to double-compartmental model (w = 1). L: -tetrahydropalmatine orally disintegrating tablets were palatable, well-tolerated, disintegrated and absorbed quickly. PMID- 21633915 TI - Tool-use to obtain honey by chimpanzees at Bulindi: new record from Uganda. AB - Honey-gathering from bee nests has been recorded at chimpanzee (Pan troglodytes) study sites across tropical Africa. Different populations employ different strategies, ranging from simple 'smash-and grab' raids to use of sophisticated tool-sets, i.e., two or more types of tool used sequentially in a single task. In this paper I present evidence of tool-use, and the probable use of a tool-set, for honey-gathering by unhabituated chimpanzees at Bulindi, a forest-farm mosaic south of the Budongo Forest in Uganda. Between June and December 2007, 44 stick tools were found in association with 16 holes dug in the ground, corresponding to the period when stingless bees (Meliponula sp.) appeared in chimpanzee dung. In 11 cases the confirmed target was a Meliponula ground nest. Two potential tool types were distinguished: digging sticks encrusted with soil, and more slender and/or flexible sticks largely devoid of soil that may have functioned to probe the bees' narrow entry tubes. Reports of chimpanzees using tools to dig for honey have been largely confined to Central Africa. Honey-digging has not previously been reported for Ugandan chimpanzees. Similarly, use of a tool-set to obtain honey has thus far been described for wild chimpanzee populations only in Central Africa. Evidence strongly suggests that Bulindi chimpanzees also use sticks in predation on carpenter bee (Xylocopa sp.) nests, perhaps as probes to locate honey or to disable adult bees. These preliminary findings from Bulindi add to our understanding of chimpanzee technological and cultural variation. However, unprotected forests at Bulindi and elsewhere in the region are currently severely threatened by commercial logging and clearance for farming. Populations with potentially unique behavioral and technological repertoires are being lost. PMID- 21633916 TI - Soluble TNF-alpha receptor I encoded on plasmid vector and its application in experimental gene therapy of radiation-induced lung fibrosis. AB - Post-radiation inflammatory reaction leads to an irreversible pulmonary fibrosis which may cause lethal respiratory insufficiency. Pathological inflammatory and fibrotic changes might be attenuated by inhibiting tumour necrosis factor (TNF) alpha activity using TNF-alpha soluble receptors. Thus, an experimental antifibrotic gene therapy with the plasmid vector encoding a mouse soluble receptor I for TNF-alpha (psTNFR-I) was assessed. Soluble TNFR-I encoding gene was cloned into pcDNA3.1 plasmid. The ability of psTNFR-I expressing vector to transfect cells, and its biological activity in vitro and in vivo were examined by PCR, RT-PCR, MTT assay and ELISA. The C57Bl/6J mice received single intramuscular injection of psTNFR-I, conjugated with polyetylenimine (PEI) 25 kDa, equally divided to both hind legs, 3 days before irradiation (20 Gy, Co60), and either a single injection or ten injections once a week after irradiation. The data proved the effectiveness of psTNFR-I product to neutralise TNF-alpha activity in vitro. The in vivo plasmid incorporation and maintenance was confirmed. Measurements of plasma soluble TNFR-I levels showed that the in vivo gene transfer was effective. PEI was found to enhance transfection efficiency in vivo. The psTNFR-I/PEI complexes caused no toxicity in the transfected mice. C57Bl/6J mice that received prolonged psTNFR-I/PEI injections developed lethal fibrotic syndrome and died 8 weeks later than the mice treated with a double plasmid injection and the control mice treated with a control plasmid. Sequential administration of soluble TNFR-I by a nonviral, intramuscular gene transduction in the early and late post-radiation inflammatory phase prolonged survival of irradiated mice and attenuated the symptoms of lung fibrosis. The psTNFR-I gene transduction may provide a safe and simple method to partially neutralise TNF alpha activity and prevent radiation-induced lung injury. PMID- 21633917 TI - Prevention of lyme disease: promising research or sisyphean task? AB - Borrelia burgdorferi sensu lato (Spirochaetes) is a group of at least 12 closely related species, some of which are responsible for chronic zoonotic infection that may cause Lyme disease. The only experimentally confirmed vector transmitting Borrelia to mammals is the Ixodes ticks. Borrelia is a highly adapted pathogen that can survive in the host organism in spite of the intense immune responses. Some patients have chronic long-lasting complications despite antibiotic therapy, probably due to adverse effects of the immune responses. A preventive vaccine against this bacterium has not been available due to the relatively broad spectrum and antigenic variability of Borrelia-surface lipoproteins and the different epitope recognition by experimental animals and humans. Although a human vaccine was marketed in the USA, it has been already pulled off the market. In addition, this vaccine was effective only in the USA, where the only pathogenic species is B. burgdorferi sensu stricto. Recent data indicate that a broadly effective vaccine will to be composed of a mixture of several antigens or multiple epitopes. PMID- 21633918 TI - Increased frequency of CD4+CD25(high)FoxP3+ regulatory T cells in patients with hepatocellular carcinoma. AB - Accumulating evidence suggests regulatory T cells (Tregs) are associated with impaired antitumor responses. However, the relationship between the CD4(+)CD25(high)FoxP3(+) Treg and hepatocellular carcinoma (HCC) has not been well investigated. Levels of CD4(+)CD25(high)FoxP3(+) Tregs in peripheral blood mononuclear cells (PBMCs) from HCC patients and healthy donors, tumor infiltrating lymphocytes (TILs) extracted from HCC, and hepatic lymphocytes extracted from resected liver were measured by flow cytometry, and their effects on T-cell proliferation was determined by (3)H-thymidine incorporation. Serum levels of interleukin (IL)-10 and transforming growth factor (TGF)-beta1 were measured by enzyme linked immunosorbent assay. The frequency of Tregs in PBMCs from HCC patients was higher than that from healthy donors. Similarly, the frequency of Tregs in TILs was higher than that of hepatic lymphocytes. On the other hand, the (3)H-thymidine uptake by TILs and PBMCs from HCC patients was decreased drastically when compared to the counterparts from normal controls. Furthermore, serum IL-10 and TGF-beta1 levels increased significantly in HCC patients when compared to the healthy donors. This study identified an increased frequency of CD4(+)CD25(high)FoxP3(+) Tregs in patients with HCC. The elevated serum IL-10, TGF-beta1 levels also correlated with impaired antitumor responses in these patients. Further effort is needed to establish new immunotherapeutic strategies designed to modulate Tregs to promote a competent antitumor response. PMID- 21633919 TI - Bim: guardian of tissue homeostasis and critical regulator of the immune system, tumorigenesis and bone biology. AB - One of the most important roles of apoptosis is the maintenance of tissue homeostasis. Impairment of apoptosis leads to a number of pathological conditions. In response to apoptotic signals, various proteins are activated in a pathway and signal-specific manner. Recently, the pro-apoptotic molecule Bim has attracted increasing attention as a pivotal regulator of tissue homeostasis. The Bim expression level is strictly controlled in both transcriptional and post transcriptional levels. This control is dependent on cell, tissue and apoptotic stimuli. The phenotype of Bim-deficient mice is a systemic lupus erythematosus like autoimmune disease with an abnormal accumulation of hematopoietic cells. Bim is thus a critical regulator of hematopoietic cells and immune system. Further studies have revealed the critical roles of Bim in various normal and pathological conditions, including bone homeostasis and tumorigenesis. The current understanding of Bim signaling and roles in the maintenance of tissue homeostasis is reviewed in this paper, focusing on the immune system, bone biology and tumorigenesis to illustrate the diversified role of Bim. PMID- 21633920 TI - Evaluating coalition capacity to strengthen community-academic partnerships addressing cancer disparities. AB - The Arkansas Cancer Connection Program is a community-academic partnership between the University of Arkansas for Medical Sciences and nine community-based coalitions designed to address cancer health disparities through community-based participatory research. In 2005, a survey measuring coalition capacity was administered to 51 Cancer Council members to assess training needs and increase coalition capacity. The highest scoring components were leadership and member engagement while the lowest were development and capacity effectiveness. Effectiveness correlated with aspects of coalition capacity. The evaluation identified training needs, which were met by projects leveraging the coalition's strengths to advance community-based participatory research addressing cancer disparities. PMID- 21633921 TI - Transplantation of progenitor cells and regeneration enhancement in acute myocardial infarction (TOPCARE-AMI): final 5-year results suggest long-term safety and efficacy. AB - BACKGROUND: Limited data is available for investigating the long-term safety and effects of intracoronary progenitor cell therapy in patients with acute myocardial infarction (AMI). OBJECTIVE: To assess the clinical course, NT-proBNP and MRI data as objective markers of cardiac function of the TOPCARE-AMI patients at 5-year follow-up. DESIGN: The TOPCARE-AMI trial was the first randomized study investigating the effects of intracoronary infusion of circulating (CPC) or bone marrow-derived progenitor cells (BMC) in 59 patients with successfully reperfused AMI. RESULTS: Five-year follow-up data were completed in 55 patients, 3 patients were lost to follow-up. None of the patients showed any signs of intramyocardial calcification or tumors at 5 years. One patient died during the initial hospitalization, no patient was rehospitalized for heart failure and 16 patients underwent target vessel revascularization (TVR). Only two TVRs occurred later than 1 year after cell administration making it very unlikely that the infused cells accelerate atherosclerotic disease progression. Serum levels of NT-proBNP remained significantly reduced at the 5-year follow-up indicating the absence of heart failure. MRI subgroup analysis in 31 patients documented a persistent improvement of LV ejection fraction (from 46 +/- 10% at baseline to 57 +/- 10% at 5 years, p < 0.001)). Simultaneously, there was a reduction (p < 0.001) in functional infarct size measured as late enhancement volume normalized to LV mass. However, whereas LV end-systolic volume remained stable, LV end-diastolic volume increased significantly. CONCLUSIONS: The 5-year follow-up of the TOPCARE AMI trial provides reassurance with respect to the long-term safety of intracoronary cell therapy and suggests favorable effects on LV function. PMID- 21633923 TI - PCR test for detecting Taenia solium cysticercosis in pig carcasses. AB - Polymerase chain reaction (PCR) test was employed to detect Taenia solium DNA in muscle lesions for validation of the meat inspection results of slaughtered pigs. Two sets of oligonucleotide primers, one targeted against the large subunit rRNA gene (TBR primers) and the other targeted against cytochrome c oxidase subunit 1 gene (Cox1 primers) of T. solium were used in this study. On reactivity in PCR test, the TBR primers and the Cox1 primers yielded products of 286 and 984 bp, respectively, in cysticercosis positive cases. Both the sets of primers were found to be highly specific, since they did not yield any PCR product in negative controls. A total of 225 pig carcasses were screened for cysticercosis by meat inspection, out of which 25 carcasses with visible cysts (16 viable and 9 degenerated cysts) were also confirmed to be positive for cysticercosis in PCR test. However, out of the 35 carcasses with suspected lesions on meat inspection, only two were found to be positive for cysticercosis in PCR test. The detection limits for both the primer sets were analyzed. The TBR primer set could detect up to 10 pg of cysticercus DNA, whereas the Cox1 primer set could detect only up to 1 ng. It is evident from the study that PCR test is an efficient tool for validation of meat inspection results and also to rule out ambiguity in carcass judgment of suspected cases of porcine cysticercosis. PMID- 21633924 TI - Reversible posterior leukoencephalopathy syndrome and trastuzumab. AB - Reversible posterior leukoencephalopathy syndrome (RPLS) is a serious condition that manifests as headache, convulsions, visual disturbance, and a characteristic magnetic resonance image (MRI) of the brain. We now describe a case of RPLS that was likely attributable to trastuzumab, a monoclonal antibody against human epidermal growth factor receptor-2 (HER2). Accumulating evidence has shown that molecular targeted agents, especially those with antiangiogenic activity cause significant hypertension which can lead to development of RPLS. Trastuzumab is also shown to inhibit tumor angiogenesis by decreasing the production of VEGF and activating antiangiogenic factors. In a clinical trial of trastuzumab, adverse effects of trastuzumab include hypertension, even though it is low incidence (~10%). Although RPLS is potently reversible, it may result in an irreversible brain damage without prompt appropriate treatment. Given the increasing use of trastuzumab in patients with breast cancer, gastric cancer, or other solid tumors, physicians should be aware of this syndrome associated with acute hypertension during trastuzumab treatment. PMID- 21633925 TI - Enhanced cell cycle perturbation and apoptosis mediate the synergistic effects of ST1926 and ATRA in neuroblastoma preclinical models. AB - Retinoic acid therapy is nowadays an important component of treatment for residual disease of stage IV neuroblastoma after multimodal therapy. Nevertheless, arising resistance and treatment toxicity could represent relevant limiting factors. In the present study, we show that retinoic acid enhances the cytostatic and apoptogenic properties of the novel adamantyl retinoid ST1926 in a panel of neuroblastoma cells with different p53 status and caspase 8 expression, resulting in synergistic effects as assessed by Combination Index and Isobologram analysis. Under conditions where the two drugs alone produced no toxic effects, their combination resulted in enhanced G2-M arrest and sub-G1 population as shown by BrdU pulse-chase and labeling experiments. PARP cleavage, caspase 3, 8 and 9 activation and modulation of DR4 and FAS were indicative of enhanced apoptosis triggered by the co-incubation of the two drugs whereas neither ST1926-mediated genotoxic damage nor ATRA-differentiating effects were affected by the combined treatment. Caspase-3 and 8-mediated apoptosis appeared to play an important role in the drugs synergism. In fact, the addition of a pan-caspase inhibitor ZVAD-FMK reverted this effect in SK-N-DZ cells, and synergism was confined to limited drugs doses in HTLA cells not expressing caspase-8. Although not modulated, p53 appeared to enhance cells responsiveness to retinoid/ATRA combination. In vivo studies in the most sensitive neuroblastoma model SK-N-DZ, confirmed enhanced activity of the drugs combination vs single treatments. The study provides important lines of evidence that such a drugs combination could represent a less toxic and more effective approach for maintenance treatment in children with neuroblastoma. PMID- 21633926 TI - Oxidative stress, cytokine/chemokine and disruption of blood-brain barrier in neonate rats after meningitis by Streptococcus agalactiae. AB - We verify the levels of cytokine/chemokine, myeloperoxidase activity, oxidative stress and disruption of BBB in hippocampus and cortex of the neonate Wistar rats after meningitis by S. agalactiae. In the hippocampus the levels were increased of CINC-1 at 6 h and 12 h, IL-1beta at 6, 12 and 24 h, IL-6 at 6, 24 and 96 h, IL 10 at 24, 48 and 96 h and TNF-alpha at 24 h and 96 h. In the cortex the CINC-1 and IL-1beta levels were found increased at 6 h. The MPO activity was significantly elevated at 24, 48 and 98 h in hippocampus and at 6, 12, 24, 48 and 96 h in the cortex. The breakdown of BBB started at 12 h.TBARS levels were elevated in the hippocampus at 6, 12, 24, 48, 72 and 96 h and cortex at 72 and 96 h. Protein carbonyls were elevated in the hippocampus and cortex at 6, 24, 48, 72 and 96 h. There was a decrease of SOD activity in hippocampus and in cortex. Catalase activity was elevated in hippocampus at 6 h and in the cortex at 12 and 96 h. Neonatal bacterial infections of the CNS are severe, the interference with the complex network of cytokines/chemokine, other inflammatory mediators and oxidants tend to aggravate the illness and can be involved in the breakdown of the BBB. PMID- 21633927 TI - Burden of osteoporosis in adults in Korea: a national health insurance database study. AB - We evaluated the number of osteoporosis patients under treatment and secular trends in 2005-2008 in South Korea. We investigated nationwide data regarding the number of osteoporosis patients under treatment in South Korea using data from the Health Insurance Review and Assesment Service (HIRA), which includes nationwide information [corrected]. Reimbursement records from the HIRA database between 1 January 2004 and 31 December 2008 were investigated. Patients aged >=30 years old with osteoporosis were identified based on a study-defined algorithm using prescription data and diagnostic codes. During the study periods, the number of patients receiving medical treatment related to osteoporosis increased from 1,034,399 to 1,392,189 for women and from 120,496 to 171,902 for men. The calculated proportion of osteoporosis patients under treatment in the general population over 50 years of age was 6.1% for men and 33.3% for women, and in the general population over 30 years of age was 2.7% for men and 16.6% for woman. More than 40% of patients (59.1% for women; 41.2% for men) were treated with medication indicated only for osteoporosis. About 4-7% of osteoporosis patients had a past medical history suggesting a secondary cause of osteoporosis. More than 80% of all osteoporosis patients were women older than 50 years, reflecting the pronounced burden of osteoporosis among postmenopausal women. This study demonstrated a substantial increasing trend in medical claims related to osteoporosis in 2005-2008 among adults in Korea and a pronounced burden of osteoporosis among postmenopausal women. PMID- 21633928 TI - Serotonin transporter genotype moderates the link between children's reports of overprotective parenting and their behavioral inhibition. AB - The goal of the current study was to examine environmental and genetic correlates of children's levels of behavioral inhibition (BI). Participants were 100 mother child pairs drawn from the community who were part of a larger study of the intergenerational transmission of depression. Results indicated that higher levels of maternal overprotection, as reported by the child, were associated with elevations in BI among children carrying two copies of the lower expressing 5 HTTLPR alleles (S or L(G)), but not among those carrying only one copy or those homozygous for the L(A) allele. In addition, this interaction was specific for the social component of BI, not the nonsocial component. This relation was maintained even after statistically controlling for children's and mother's psychopathology. Together, these findings add to emerging research demonstrating that G * E interactions predict variation in BI during childhood. PMID- 21633929 TI - Potential dopamine-1 receptor stimulation in hypertension management. AB - The role of dopamine receptors in blood pressure regulation is well established. Genetic ablation of both dopamine D1-like receptor subtypes (D1, D5) and D2-like receptor subtypes (D2, D3, D4) results in a hypertensive phenotype in mice. This review focuses on the dopamine D1-like receptor subtypes D1 and D5 (especially D1 receptors), as they play a major role in regulating sodium homeostasis and blood pressure. Studies mostly describing the role of renal dopamine D1-like receptors are included, as the kidneys play a pivotal role in the maintenance of sodium homeostasis and the long-term regulation of blood pressure. We also attempt to describe the interaction between D1-like receptors and other proteins, especially angiotensin II type 1 and type 2 receptors, which are involved in the maintenance of sodium homeostasis and blood pressure. Finally, we discuss a new concept of renal D1 receptor regulation in hypertension that involves oxidative stress mechanisms. PMID- 21633930 TI - Infectious serologies and autoantibodies in hepatitis C and autoimmune disease associated mixed cryoglobulinemia. AB - Mixed cryoglobulinemia (MC) syndrome is an immune complex-mediated vasculitis characterized by the clinical triad of purpura, weakness, and arthralgias, the morbidity of which is mainly related to kidney and peripheral nervous system dysfunction as well as to the development of a secondary lymphoma (Ferri et al. Autoimmun Rev 7:114-120, 2007, Lidar et al. Ann N Y Acad Sci 1173:649-657, 2009, Trejo et al. Semin Arthritis Rheum 33:19-28, 2003). MC is associated with infectious and systemic disorders, principally autoimmune and lymphoproliferative diseases. Since the 1990s, a striking association (>90%) between MC and hepatitis C virus (HCV) infection has been established (Ferri and Bombardieri 2004; Pascual et al. J Infect Dis 162:569-570, 1990). However, information regarding the etiopathogenesis of HCV-negative MC is scant (Mascia et al. Dig Liver Dis 39:61 64, 2007). We hereby present our findings, as well as previously published data, regarding the presence of antibodies against infectious agents and autoantibodies in patients with MC in an attempt to establish novel associations which may shed light on the etiopathogenesis of this disease. PMID- 21633931 TI - Secretory carrier membrane proteins. AB - Secretory carrier membrane proteins (SCAMPs) are a family of integral membrane proteins that play roles in mediating exocytosis in animal cells. However, relatively little is known about the subcellular localization, trafficking, and function of SCAMPs in plants. Several recent studies in plant cells indicate that plant SCAMPs share many similarities with their mammalian homologs although there are differences. In this review, we will first summarize and compare animal and plant SCAMPs in terms of their subcellular localization, trafficking, and possible functions. We will then present a phylogenetic analysis of plant and animal SCAMPs. Finally, we will present expression analysis on selective Arabidopsis SCAMPs in the hope of pointing to directions for functional characterization of plant SCAMPs in the future. PMID- 21633932 TI - Effects of hexavalent chromium on microtubule organization, ER distribution and callose deposition in root tip cells of Allium cepa L. AB - The subcellular targets of hexavalent chromium [Cr(VI)] were examined in Allium cepa root tips with confocal laser scanning microscopy. Cr(VI) exerted dose- and time-dependent negative effects on root growth rate, the mitotic index and microtubule (MT) organization during cell division cycle. Interphase MTs were more resistant than the mitotic ones, but when affected they were shorter, sparse and disoriented. The preprophase band of MTs became poorly organized, branched or with fragmented MTs, whilst neither a perinuclear array nor a prophase spindle was formed. Metaphase spindles converged to eccentric mini poles or consisted of dissimilar halves and were unable to correctly orient the chromosomes. Anaphase spindles were less disturbed, but chromatids failed to separate; neither did they move to the poles. At telophase, projecting, lagging or bridging chromosomes and micronuclei also occurred. Phragmoplasts were unilaterally developed, split, located at unexpected sites and frequently dissociated from the branched and misaligned cell plates. Chromosomal aberrations were directly correlated with MT disturbance. The morphology and distribution of endoplasmic reticulum was severely perturbed and presumably contributed to MT disassembly. Heavy callose apposition was also induced by Cr(VI), maybe in the context of a cellular defence reaction. Results indicate that MTs are one of the main subcellular targets of Cr(VI), MT impairment underlies chromosomal and mitotic aberrations, and MTs may constitute a reliable biomonitoring system for Cr(VI) toxicity in plants. PMID- 21633933 TI - [Gunshot and splinter wounds in the head and neck area]. PMID- 21633934 TI - Hired hand or boss? AB - Deciding whether to open your own practice or work for someone else is a big decision every young physician has to face. Many start out with established medical groups, but they usually don't remain there as employees. TMA has tools and services to meet the needs of physicians just starting out independently or as employees. PMID- 21633935 TI - Good for Texas, good for u.s. AB - The way U.S. Rep. Michael Burgess, MD (R-Texas), sees it, the 2003 Texas medical liability reforms are so successful that the rest of the country should follow suit. That is why a number of Texas members of Congress - both Republican and Democrat - are either authoring or cosponsoring legislation to pass national legislative reforms similar to those in Texas or, at least, to ensure that no federal action could negate state-level reforms. PMID- 21633936 TI - Sensitive subjects. AB - Texas Medical Association policy directs physicians who treat adolescents to give them counseling and treatment or a referral on sexual development, sexually transmitted disease, birth control, pregnancy, and substance abuse. TMA, the American Medical Association, and other organizations have created materials and resources on adolescent health care and healthy lifestyles for physicians to use in their practice and professional activities. PMID- 21633937 TI - Getting the hassles out. AB - The Texas Medical Association's Hassle Factor Log is an important tool that any TMA member can access. TMA Payment Advocacy Department staff follow up on Hassle Factor Log complaints to help physicians resolve payment issues with health plans. Since 2009, the Hassle Factor Log program has helped physician practices recover more than $1.1 million in unpaid claims. PMID- 21633938 TI - Playing house. AB - Small group, problem-based learning sessions began at The University of Texas Medical Branch in the late 1990s when administrators overhauled the school's curriculum. The new curriculum was intended to shift medical education away from passive learning and toward active learning. The results have been dramatic. Students' mean scores on Step 1 of the U.S. Medical Licensing Exam had been lagging behind the national mean but are now substantially higher than the mean. PMID- 21633939 TI - Take it outside. AB - Hiring an outside billing company can increase a practice's collections. Medical billing companies offer data entry, monitoring and review of explanations of benefits, patient billing, management of accounts receivable, and follow-up on unpaid claims and patient balances. Doing some research on the front end can help physicians choose a legitimate company and avoid billing and collections headaches down the road. PMID- 21633940 TI - Basic techniques in human mesenchymal stem cell cultures: differentiation into osteogenic and adipogenic lineages, genetic perturbations, and phenotypic analyses. AB - This unit describes basic techniques in human mesenchymal stem cell (hMSC) cultures. It includes protocols for the differentiation of hMSCs into osteogenic and adipogenic lineages, genetic perturbations, and phenotypic analyses. hMSCs can be differentiated with dexamethasone and beta-glycerophosphate into mineralizing osteoblasts within 2 to 3 weeks, or with dexamethasone, indomethacin, and 3-isobutyl-1-methylxanthine into lipid vesicle-containing adipocytes within 1 to 2 weeks. Phenotypic changes during those highly dynamic differentiation processes can be detected by biochemical and histological assays and gene expression analyses of differentiation markers. In addition, this unit describes an electroporation method that allows the transient genetic perturbation of hMSCs. PMID- 21633941 TI - The importance of biological databases in biological discovery. AB - Biological databases play a central role in bioinformatics. They offer scientists the opportunity to access a wide variety of biologically relevant data, including the genomic sequences of an increasingly broad range of organisms. This unit provides a brief overview of major sequence databases and portals, such as GenBank, the UCSC Genome Browser, and Ensembl. Model organism databases, including WormBase, the Arabidopsis Information Resource (TAIR), and those made available through the Mouse Genome Informatics (MGI) resource are also covered. Non-sequence-centric databases, such as Online Mendelian Inheritance in Man (OMIM), the Protein Data Bank (PDB), MetaCyc, and the Kyoto Encyclopedia of Genes and Genomes (KEGG) are also discussed. PMID- 21633942 TI - Searching NCBI databases using Entrez. AB - One of the most widely used interfaces for the retrieval of information from biological databases is the NCBI Entrez system. Entrez capitalizes on the fact that there are pre-existing, logical relationships between the individual entries found in numerous public databases. The existence of such natural connections, mostly biological in nature, argued for the development of a method through which all the information about a particular biological entity could be found without having to sequentially visit and query disparate databases. Two basic protocols describe simple, text-based searches, illustrating the types of information that can be retrieved through the Entrez system. An alternate protocol builds upon the first basic protocol, using additional, built-in features of the Entrez system, and providing alternative ways to issue the initial query. The support protocol reviews how to save frequently issued queries. Finally, Cn3D, a structure visualization tool, is also discussed. PMID- 21633943 TI - Metabolomic data processing, analysis, and interpretation using MetaboAnalyst. AB - MetaboAnalyst is a comprehensive, Web-based tool designed for processing, analyzing, and interpreting metabolomic data. It handles most of the common metabolomic data types including compound concentration lists, spectral bin lists, peak lists, and raw MS spectra. In addition to providing a variety of data processing and normalization procedures, MetaboAnalyst supports a number of data analysis tasks using a range of univariate, multivariate, and machine-learning methods. MetaboAnalyst also offers two newly developed approaches-Metabolite Set Enrichment Analysis (MSEA) and Metabolic Pathway Analysis (MetPA)-for metabolomic data interpretation. MSEA helps detect biologically meaningful metabolite sets that have been enriched in human metabolomic studies, while MetPA allows users to identify any metabolic pathways that have been perturbed. MetaboAnalyst enables facile interactive exploration and visualization of nearly all of its results. At the end of each session, it produces a detailed analysis report with graphical, tabular, and textual output that summarizes each analytical method used and each result generated. PMID- 21633944 TI - An introduction to recognizing functional domains. AB - This unit provides an overview of issues involved in domain recognition in protein and DNA sequences. It opens with a discussion of the two primary methods of domain representation, namely consensus sequences and alignment matrices (e.g., the log-odds matrix). The unit continues with a brief overview of some of the resources available for identifying functional domains in nucleotide sequences (e.g., transcription factor binding sites). In addition, it reviews databases such as Pfam and InterPro, which are available for protein analysis. PMID- 21633945 TI - Using MACS to identify peaks from ChIP-Seq data. AB - Model-based Analysis of ChIP-Seq (MACS) is a command-line tool designed by X. Shirley Liu and colleagues to analyze data generated by ChIP-Seq experiments in eukaryotes, especially mammals. MACS can be used to identify transcription factor binding sites and histone modification-enriched regions if the ChIP-Seq data, with or without control samples, are given. This unit describes two basic protocols that provide detailed information on how to use MACS to identify either the binding sites of a transcription factor or the enriched regions of a histone modification with broad peaks. Furthermore, the basic ideas for the MACS algorithm and its appropriate usage are discussed. PMID- 21633947 TI - Polymorphisms in the toll-like receptor 9 gene associated with sepsis and multiple organ dysfunction after major blunt trauma. AB - BACKGROUND: Toll-like receptor (TLR) 9 is the pattern recognition receptor for microbial DNA. Genetic variation within pattern recognition receptors for bacterial endotoxin and exotoxin has been shown to be associated with the risk of sepsis and organ dysfunction in critical illness. However, little is known about the clinical relevance of TLR9 gene polymorphisms in critical illness. METHODS: A total of 557 patients with major blunt trauma were included in the study. Genetic variation data for the entire TLR9 gene were obtained from the HapMap Project. The genotypes of TLR9 gene polymorphisms were determined using a pyrosequencing method. Whole peripheral blood samples obtained immediately after admission were stimulated with bacterial DNA and production of tumour necrosis factor (TNF) alpha was then determined. Sepsis morbidity rate and multiple organ dysfunction (MOD) scores were assessed. RESULTS: Of five single-nucleotide polymorphisms (SNPs) genotyped, four (rs187084, rs352139, rs352140 and rs352162) existed as common SNPs and were in strong linkage disequilibrium. Both rs187084 and rs352162 were significantly associated with TNF-alpha production by peripheral blood leucocytes in response to bacterial DNA stimulation and a higher sepsis morbidity rate in patients with major trauma. In addition, the rs352162 polymorphism was significantly associated with MOD scores, whereas rs187084 showed a trend to be associated with MOD score. CONCLUSION: TLR9 polymorphisms rs187084 and rs352162 might be used to provide relevant risk estimates for the development of sepsis and MOD in patients with major trauma. PMID- 21633948 TI - Five-year results of a randomized clinical trial comparing endovenous laser ablation with cryostripping for great saphenous varicose veins. AB - BACKGROUND: This was the long-term follow-up of a previously reported randomized clinical trial comparing endovenous laser ablation (EVLA) with cryostripping for great saphenous varicose veins. METHODS: A total of 120 patients with great saphenous varicose veins were randomized 1:1 to EVLA or cryostripping. Principal outcome measures were freedom from incompetence or neovascularization on duplex imaging, and improvement in Venous Clinical Severity Score (VCSS) and Aberdeen Varicose Vein Severity Score (AVVSS) 5 years after treatment. RESULTS: Life-table analysis showed freedom from duplex-derived incompetence and neovascularization at 5 years in 62 (95 per cent confidence interval 50 to 76) per cent after EVLA and in 51 (39 to 66) per cent after cryostripping (P = 0.246). Neovascularization was more common after cryostripping, but incompetent tributaries were more common after EVLA. VCSS and AVVSS values improved significantly after treatment in both groups, and were maintained for 5 years, but with no significant difference between the groups. CONCLUSION: In this study, no significant difference was demonstrated in late outcome after EVLA or cryostripping in patients with great saphenous varicose veins. PMID- 21633949 TI - Outcomes following a limited approach to radiotherapy in rectal cancer. AB - BACKGROUND: Variation in the use of neoadjuvant and adjuvant radiotherapy for rectal cancer suggests an opportunity to avoid it in all but patients at highest risk of local recurrence. METHODS: Between 1 July 1999 and 1 February 2006, patients with primary rectal cancer were treated by a single surgeon operating at McMaster University, Hamilton, Ontario, Canada. Digital rectal examination and pelvic computed tomography were used to determine whether the mesorectal margin was threatened by tumour and thus whether preoperative radiotherapy would be needed. The study outcome was local tumour recurrence. RESULTS: Forty-six (48 per cent) of 96 patients received preoperative radiation therapy. The median follow up was 4.2 years. Tumours were fixed or tethered in 31 (67 per cent) of the 46 irradiated patients. In contrast, no tumour was fixed in unirradiated patients and only ten (20 per cent) of the 50 tumours were tethered. The proportion of patients with stage I or II tumours based on final pathology was similar: 61 per cent (28 of 46) and 56 per cent (28 of 50) in irradiated and unirradiated groups respectively (P = 0.287). There were four (9 per cent) and two (4 per cent) local recurrences among irradiated and unirradiated patients respectively (P = 0.422). CONCLUSION: Limiting preoperative radiotherapy in rectal cancer to patients with a threatened circumferential margin does not compromise patient outcome. PMID- 21633950 TI - Duration of hepatic vascular inflow clamping and survival after liver resection for hepatocellular carcinoma. AB - BACKGROUND: The aim of this study was to evaluate the influence of the duration of hepatic vascular inflow clamping (Pringle time) on the survival of patients with any type of liver background (not only cirrhosis) undergoing liver resection for hepatocellular carcinoma (HCC). METHODS: Patients who underwent liver resection between April 2000 and December 2008 for HCC using the Pringle manoeuvre were identified retrospectively from an institutional database and divided into two groups: group 1 had a Pringle time of 60 min or less, and group 2 a Pringle time of more than 60 min. Univariable and multivariable analyses were performed to identify predictors of postoperative survival. Kaplan-Meier analysis was used to compare overall survival between the groups. RESULTS: A total of 357 patients were enrolled; 242 patients had a Pringle time of 60 min or less (group 1), and 115 patients had a Pringle time of more than 60 min (group 2). Patients in group 2 had a shorter overall survival than those in group 1 (P = 0.010). Univariable analyses showed that type of HCC (primary versus recurrent), maximum tumour diameter, hepatic venous infiltration, platelet count, serum protein induced by vitamin K absence or antagonist II level, blood loss (700 ml or less versus more than 700 ml), duration of operation (300 min or less versus more than 300 min) and Pringle time (60 min or less versus more than 60 min) were predictive of postoperative survival. Multivariable analysis indicated that only Pringle time was associated with postoperative survival (odds ratio 1.83, 95 per cent confidence interval 1.08 to 3.10; P = 0.024). CONCLUSION: Longer Pringle time is an important predictor of shorter postoperative survival in patients undergoing liver resection for HCC. PMID- 21633951 TI - The significant rectal neoplasm and mucosectomy by transanal endoscopic microsurgery. PMID- 21633952 TI - Operative notes do not reflect reality in laparoscopic cholecystectomy. AB - BACKGROUND: Operative notes represent an essential element in safe patient care and should therefore be clear and accurate. This comparative study examined whether operative notes accurately represented the laparoscopic cholecystectomy (LC) as performed. METHODS: Nine Dutch teaching and non-teaching hospitals were invited to record 20 successive LCs each and to collect the corresponding operative notes. The main outcome measures were overall differences and correspondence between video recordings and notes based on the Dutch guideline for LC and the occurrence of iatrogenic gallbladder perforation. A comparison was made of the cumulative results of recordings and operative notes, and individual recordings were compared with the corresponding notes. RESULTS: Seven hospitals participated in the study; 125 video recordings and operative notes were fully analysed. Recordings showed more steps of the procedure than did notes. Individual comparisons showed significant differences (P<=0.001) between the recording and the corresponding note for the steps 'Introducing trocars under vision', 'Condition of the gallbladder', 'Critical view of safety' and 'Removing first and second trocar under vision'. Iatrogenic gallbladder perforation with spilled bile occurred in 31 patients (24.8 per cent), and was both recorded and reported in 29 patients. Iatrogenic gallbladder perforation with spilled bile and spilled stones occurred in 15 patients (12.0 per cent), and was recorded and reported in 11 patients. CONCLUSION: Operative notes do not adequately represent the actual LCs performed as they describe fewer important procedural steps. It is suggested that operative notes should include video recordings. PMID- 21633953 TI - MicroRNA-30a inhibits epithelial-to-mesenchymal transition by targeting Snai1 and is downregulated in non-small cell lung cancer. AB - MicroRNAs (miRNAs) are small non-coding RNAs which regulate gene expression by base-pairing to the 3'-UTR of the target mRNA. Recently, miRNAs have been shown to regulate cancer metastasis, however, central molecular mechanisms of this ability still need to be investigated. Epithelial to mesenchymal transition (EMT), which is characterized especially by repression of E-cadherin expression and increased cell motility, is an essential component of cancer metastasis and progression. In the present study, we found that Snai1, a known transcriptional repressor of E-cadherin and modulator of EMT, is post-transcriptionally targeted by miRNA-30a in non-small cell lung cancer (NSCLC). Consistent with this, microRNA-30a expression was found inversely proportional to the invasive potential of various NSCLC cell lines, correlating positively with E-cadherin (epithelial marker) and negatively with N-cadherin (mesenchymal marker) expression. Forced re-introduction of miR-30a significantly altered cell morphology, in vitro invasion and migration of invasive cell lines, this being paralleled by a downregulation of Snai1 and upregulation of E-cadherin expression. Using a chicken embryonic metastasis assay, we found that miR-30a suppresses in vivo distant metastasis to the lungs and liver. Finally, we screened the expression of miR-30a in 64 consecutively resected NSCLC patients and found that, in 81% of the patients, expression of miR-30a was downregulated significantly (p < 0.0001) in tumors compared to corresponding normal tissues. These results suggest that miR-30a targets Snai1, inhibits invasion and metastasis, and is downregulated in NSCLC. PMID- 21633954 TI - Consequence of dose scheduling of sunitinib on host immune response elements and vaccine combination therapy. AB - Our study investigated the immunomodulatory effects of sunitinib to rationally design combinational platforms with immunotherapies for the treatment of solid tumors. Using a mouse model, we studied the effects of sunitinib given for 4 weeks at concentrations comparable to 37.5-50 mg/day in humans, followed by 2 weeks off the drug (sunitinib 4/2). We assessed the effect of differently timed combinations of sunitinib and a poxvirus-based vaccine encoding carcinoembryonic antigen (CEA) plus 3 costimulatory molecules on immune responses in CEA transgenic (CEA-Tg) mice. Antitumor studies were performed in CEA-Tg mice bearing CEA-transfected MC38 murine colon carcinomas (MC38-CEA), treated either concurrently or sequentially with sunitinib and vaccine. In vitro, sunitinib inhibited PDGFR phosphorylation on MC38-CEA cells at concentrations similar to those biologically available during human treatment. In vivo, one cycle of sunitinib 4/2 caused bimodal immune effects: (a) decreased regulatory cells during the 4 weeks of treatment and (b) an immune-suppression rebound during the 2 weeks of treatment interruption. In a model using CEA-Tg mice bearing CEA(+) tumors, continuous sunitinib followed by vaccine increased intratumoral infiltration of antigen-specific T lymphocytes, decreased immunosuppressant T regulatory cells and myeloid-derived suppressor cells, reduced tumor volumes and increased survival. The immunomodulatory activity of continuous sunitinib administration can create a more immune-permissive environment. In combination with immunotherapies, sunitinib treatment should precede vaccine, to precondition the immune system, to maximize the response to vaccine-mediated immune enhancement. PMID- 21633955 TI - ABO blood group and breast cancer incidence and survival. AB - ABO blood type has been associated with risk and survival for several malignancies; however, data for an association with breast cancer are inconsistent. Our study population consisted of Nurses' Health Study participants with self-reported serologic blood type and/or ABO genotype. Using Cox proportional hazards regression, we examined the association between serologic blood type and incident breast cancer among 67,697 women, including 3,107 cases. In addition, we examined the association with ABO genotype in a nested case control study of 1,138 invasive breast cancer cases and 1,090 matched controls. Finally, we evaluated the association between serologic blood type and survival among 2,036 participants with breast cancer. No clear association was seen between serologic blood type or ABO genotype and risk of total breast cancer, invasive breast cancer or breast cancer subtypes. Compared to women with blood type O, the age-adjusted incidence rate ratios for serologic blood type and total breast cancer were 1.06 (95% CI, 0.98-1.15) for type A, 1.06 (95% CI, 0.93-1.22) for AB and 1.08 (95% CI, 0.96-1.20) for B. In genetic analyses, odds ratios for invasive breast cancer were 1.05 (95% CI, 0.87-1.27) for A/O, 1.21 (95% CI, 0.86 1.69) for A/A, 0.84 (95% CI, 0.56-1.26) for A/B, 0.84 (95% CI, 0.63-1.13) for B/O and 1.17 (95% CI, 0.35-3.86) for B/B, compared to O/O. No significant association was noted between blood type and overall or breast cancer-specific mortality. Our results suggest no association between ABO blood group and breast cancer risk or survival. PMID- 21633956 TI - A systematic review of measures of therapeutic engagement in psychosocial and psychological treatment. AB - This article reports a systematic review of engagement measures for psychosocial therapy. MEDLINE, EMBASE, and PsycINFO databases were searched to identify English-language studies (published 1980 to February 2010) that reported on an instrument/rating scale to measure engagement in psychosocial treatment for mental health difficulties. Forty-seven studies were identified, reporting information on 40 measures of treatment engagement. Although our findings suggest that therapeutic engagement appears to be considered an important construct to assess, they also reveal that there is little consensus in the definition of engagement employed. Few measures are generalizable across treatment settings and clinical populations, and limited information is reported on the indices of reliability and validity. It is concluded that further work is required to develop adequate measures of therapeutic engagement. PMID- 21633957 TI - Forgiveness, depression, and suicidal behavior among a diverse sample of college students. AB - Depression and suicide are significant public health concerns for college-age young adults. Meaning-based characteristics, such as forgiveness, a voluntary coping process involving offering, feeling, or seeking a change from negative to positive cognitions, behaviors, and affect toward a transgressor, may buffer such poor mental health outcomes. Utilizing mediation analyses, we examined cross sectional associations between forgiveness, depression, and suicidal behavior in a diverse student sample reporting mild to severe depressive symptoms. The effect of self-forgiveness on suicidal behavior was fully mediated by depression; self forgiveness was associated with depression and, in turn, with suicidal behavior. Forgiveness of others was directly associated with suicidal behavior. Prospective research is needed, yet self and other-forgiveness may be appropriate targets for promotion in suicide prevention efforts. PMID- 21633958 TI - Construct drift in the MMPI-2 Restructured Clinical scales: further evidence and a possible historic example. AB - Construct drift (Nichols, 2006) describes the possibility that the MMPI-2 Reconstructed Clinical (RC) scales are less accurate measures of the primary traits than the original Clinical scales. The authors review the development of RC scales RC4, RC7, and RC9 and provide further evidence that these scales measure traits and behaviors that are not necessarily equivalent to those assessed by the original scales. Further, the development of the original Clinical Scale 7 is reviewed and evidence is provided that suggests that construct drift is not a new phenomenon, but may have been present in the development of the original scales. Suggestions for future research as well as cautions with regard to the clinical interpretation of the RC scales are provided. PMID- 21633959 TI - Methyl donor deficiency induces cardiomyopathy through altered methylation/acetylation of PGC-1alpha by PRMT1 and SIRT1. AB - Cardiomyopathies occur by mechanisms that involve inherited and acquired metabolic disorders. Both folate and vitamin B12 deficiencies are associated with left ventricular dysfunction, but mechanisms that underlie these associations are not known. However, folate and vitamin B12 are methyl donors needed for the synthesis of S-adenosylmethionine, the substrate required for the activation by methylation of regulators of energy metabolism. We investigated the consequences of a diet lacking methyl donors in the myocardium of weaning rats from dams subjected to deficiency during gestation and lactation. Positron emission tomography (PET), microscope and metabolic examinations evidenced a myocardium hypertrophy, with cardiomyocyte enlargement, disturbed mitochondrial alignment, lipid droplets, decreased respiratory activity of complexes I and II and decreased S-adenosylmethionine:S-adenosylhomocysteine ratio. The increased concentrations of triglycerides and acylcarnitines were consistent with a deficit in fatty acid oxidation. These changes were explained by imbalanced acetylation/methylation of PGC-1alpha, through decreased expression of SIRT1 and PRMT1 and decreased S-adenosylmethionine:S-adenosylhomocysteine ratio, and by decreased expression of PPARalpha and ERRalpha. The main changes of the myocardium proteomic study were observed for proteins regulated by PGC-1alpha, PPARs and ERRalpha. These proteins, namely trifunctional enzyme subunit alpha complex, short chain acylCoA dehydrogenase, acylCoA thioesterase 2, fatty acid binding protein-3, NADH dehydrogenase (ubiquinone) flavoprotein 2, NADH dehydrogenase (ubiquinone) 1alpha-subunit 10 and Hspd1 protein, are involved in fatty acid oxidation and mitochondrial respiration. In conclusion, the methyl donor deficiency produces detrimental effects on fatty acid oxidation and energy metabolism of myocardium through imbalanced methylation/acetylation of PGC-1alpha and decreased expression of PPARalpha and ERRalpha. These data are of pathogenetic relevance to perinatal cardiomyopathies. PMID- 21633960 TI - Always, never, or sometimes: examining variation in condom-use decision making among Black adolescent mothers. AB - Our purpose in this study was to describe Black adolescent mothers' decisions regarding condom use and non-use with their male sex partners, including their children's fathers. Research on partner type and condom use has been insufficiently focused on understanding the specific influence that the biological father of the baby has on condom use among adolescent mothers. We conducted five focus groups and three interviews with 31 predominantly African American mothers. We found that their decisions to use condoms always, never, or sometimes were based on partner type and on emotional and relationship factors. The "baby daddy" was the only partner with whom they never used condoms. HIV/STI prevention interventions for adolescent mothers must address risk taking with their children's biological fathers. PMID- 21633961 TI - Water order profiles on phospholipid/cholesterol membrane bilayer surfaces. AB - Water is pivotal in the stabilization of macromolecular biological structures, although the dynamic ensemble structure of water near to molecular surfaces has yet to be fully understood. We show, through molecular simulation and fluorescence measurements, that water at the membrane surface is substantially more ordered than bulk water, due to a loss of hydrogen bonding between water molecules, coupled with an alignment of lipid and water dipole moments. Ordering of the water leads to a gradient in the effective dielectric permittivity, which is evident in both the molecular simulations and the fluorescence measurements. A lower effective dielectric permittivity was correlated with a decreasing degree of hydrogen bonding over the same spatial range. The water molecules closest to the lipid headgroup oxygen atoms form hydrogen bonds which exhibit a mean lifetime of 6.3 ps, compared with a mean lifetime of water-water hydrogen bonds of less than 2 ps. Membranes made up purely of phosphatidylcholine (PC) were compared with those made with a PC/cholesterol ratio relevant to cell membranes. Clear differences were found between these membrane configurations. These observations point to molecular structural differences in the surface environments of membranes and may underlie regional differences in the surface biophysical properties of membrane microdomains. PMID- 21633962 TI - Are viruses a source of new protein folds for organisms? - Virosphere structure space and evolution. AB - A crucially important part of the biosphere - the virosphere - is too often overlooked. Inclusion of the virosphere into the global picture of protein structure space reveals that 63 protein domain superfamilies in viruses do not have any structural and evolutionary relatives in modern cellular organisms. More than half of these have functions which are not virus-specific and thus might be a source of new folds and functions for cellular life. The number of viruses on the planet exceeds that of cells by an order of magnitude and viruses evolve up to six orders of magnitude faster. As a result, cellular species are subject to a constitutive 'flow-through' of new viral genetic material. Due to this and the relaxed evolutionary constraints in viruses, the transfer of domains between host to-virus could be a mechanism for accelerated protein evolution. The virosphere could be an engine for the genesis of protein structures, and may even have been so before the last universal common ancestor of cellular life. PMID- 21633963 TI - The time of timing: how Polycomb proteins regulate neurogenesis. AB - The study of mammalian corticogenesis has revealed a critical role for Polycomb group (PcG) factors in timing the execution of developmental choices. Meanwhile, the study of post-translational modifications of PcG factors marks a symmetrical point, namely that the activity of PcG proteins is itself timed in a manner that links progression through the cell cycle to targeting of downstream genes. Finally, in a third symmetrical twist, the studies that dissect the timing of neural fate by Polycomb are also uncovering the importance of timing in the experimental mutation, since ablation of the same PcG member at different developmental stages yields dramatically different results. Here, I weave together these three lines of evidence and develop a unifying model that clarifies the dynamics of Polycomb function in neural development and defines the salient challenges ahead. PMID- 21633964 TI - The low cost of recombination in creating novel phenotypes: Recombination can create new phenotypes while disrupting well-adapted phenotypes much less than mutation. AB - Recombination is often considered a disruptive force for well-adapted phenotypes, but recent evidence suggests that this cost of recombination can be small. A key benefit of recombination is that it can help create proteins and regulatory circuits with novel and useful phenotypes more efficiently than point mutation. Its effectiveness stems from the large-scale reorganization of genotypes that it causes, which can help explore far-flung regions in genotype space. Recent work on complex phenotypes in model gene regulatory circuits and proteins shows that the disruptive effects of recombination can be very mild compared to the effects of mutation. Recombination thus can have great benefits at a modest cost, but we do not understand the reasons well. A better understanding might shed light on the evolution of recombination and help improve evolutionary strategies in biochemical engineering. PMID- 21633965 TI - SMT vs. TOFT. PMID- 21633966 TI - Highly sensitive LC-MS/MS-ESI method for simultaneous quantitation of albendazole and ricobendazole in rat plasma and its application to a rat pharmacokinetic study. AB - A highly sensitive and specific LC-MS/MS-ESI method was developed for simultaneous quantification of albenadazole (ABZ) and ricobendazole (RBZ) in rat plasma (50 MUL) using phenacetin as an internal standard (IS). Simple protein precipitation was used to extract ABZ and RBZ from rat plasma. The chromatographic resolution of ABZ, RBZ and IS was achieved with a mobile phase consisting of 5 m m ammonium acetate (pH 6) and acetonitrile (20:80, v/v) at a flow rate of 1 mL/min on a Chromolith RP-18e column. The total chromatographic run time was 3.5 min and the elution of ABZ, RBZ and IS occurred at 1.66, 1.50 and 1.59 min, respectively. A linear response function was established for the ranges of concentrations 2.01-2007 and 6.02-6020 ng/mL for ABZ and RBZ, respectively. The intra- and inter-day precision values for ABZ and RBZ met the acceptance as per FDA guidelines. ABZ and RBZ were stable in battery of stability studies, viz. bench-top, auto-sampler and freeze-thaw cycles. The developed assay was applied to a pharmacokinetic study in rats. PMID- 21633967 TI - Canonical notch pathway protects hepatocytes from ischemia/reperfusion injury in mice by repressing reactive oxygen species production through JAK2/STAT3 signaling. AB - Hepatic ischemia/reperfusion (I/R) injury is initiated by reactive oxygen species (ROS) accumulated during the early reperfusion phase after ischemia, but cellular mechanisms controlling ROS production and scavenging have not been fully understood. In this study, we show that blocking Notch signal by knockout of the transcription factor RBP-J or a pharmacological inhibitor led to aggravated hepatic I/R injury, as manifested by deteriorated liver function and increased apoptosis, necrosis, and inflammation, both in vitro and in vivo. Interruption of Notch signaling resulted in increased intracellular ROS in hepatocytes, and a ROS scavenger cured exacerbated hepatic I/R injury after Notch signaling blockade, suggesting that Notch signal deficiency aggravated I/R injury through increased ROS levels. Notch signal blockade resulted in down-regulation of Hes5, leading to reduced formation of the Hes5-STAT3 complex and hypophosphorylation of STAT3, which further attenuated manganese superoxide dismutase (MnSOD) expression and increased ROS and apoptosis. Indeed, overexpression of a constitutively active STAT3 rescued MnSOD expression and I/R injury-induced apoptosis in the absence of Notch signaling. Finally, forced Notch activation by ligand stimulation or Hes5 overexpression reduced intracellular ROS and protected hepatocytes from apoptosis after I/R injury through the activation of STAT3 and MnSOD expression. CONCLUSION: Notch signal protects hepatocytes from I/R injury by Hes5-dependent activation of STAT3, which activates the expression of MnSOD, leading to the scavenging of ROS. PMID- 21633968 TI - Fast fat suppression RF pulse train with insensitivity to B1 inhomogeneity for body imaging. AB - In higher-field magnetic resonance imaging scanners, a spectrally selective fat saturation radiofrequency (RF) pulse does not work well because B(1) inhomogeneity increases. An adiabatic 180 degrees pulse is used to improve nonuniform fat suppression, but requires inversion recovery time. Therefore, a new RF pulse that achieves flip angles near 90 degrees and is B(1) insensitive has been developed. The pulse consists of three sinc-shaped RF pulses with different flip angles and with different time intervals between each RF pulse. Using the Bloch equations, we analyzed the optimal combination of flip angles. Experimental results demonstrated that M(z) was maintained at less than 0.05 M(0) for a B(1) inhomogeneity of +/-35%. The optimal net flip angles was adjusted to 95 degrees by varying the time interval between RF pulses. The pulse duration was 77 ms, which is less than half of the 170-ms inversion recovery time required for the adiabatic pulse. We demonstrated excellent fat suppression for body imaging. PMID- 21633969 TI - Crystal structure of the novel PaiB transcriptional regulator from Geobacillus stearothermophilus. PMID- 21633970 TI - Crystal structure of the novel PaiA N-acetyltransferase from Thermoplasma acidophilum involved in the negative control of sporulation and degradative enzyme production. AB - GCN5-related N-acetyltransferases (GNATs) are the most widely distributed acetyltransferase systems among all three domains of life. GNATs appear to be involved in several key processes, including microbial antibiotic resistance, compacting eukaryotic DNA, controlling gene expression, and protein synthesis. Here, we report the crystal structure of a putative GNAT Ta0374 from Thermoplasma acidophilum, a hyperacidophilic bacterium, that has been determined in an apo form, in complex with its natural ligand (acetyl coenzyme A), and in complex with a product of reaction (coenzyme A) obtained by cocrystallization with spermidine. Sequence and structural analysis reveals that Ta0374 belongs to a novel protein family, PaiA, involved in the negative control of sporulation and degradative enzyme production. The crystal structure of Ta0374 confirms that it binds acetyl coenzyme A in a way similar to other GNATs and is capable of acetylating spermidine. Based on structural and docking analysis, it is expected that Glu53 and Tyr93 are key residues for recognizing spermidine. Additionally, we find that the purification His-Tag in the apo-form structure of Ta0374 prevents binding of acetyl coenzyme A in the crystal, though not in solution, and affects a chain flip rotation of "motif A" which is the most conserved sequence among canonical acetyltransferases. PMID- 21633971 TI - Conserved conformational dynamics of membrane fusion protein transmembrane domains and flanking regions indicated by sequence statistics. AB - SNARE proteins and fusogenic viral membrane proteins represent the major classes of integral membrane proteins that mediate fusion of eukaryotic lipid bilayers. Although both classes have different primary structures, they share a number of basic architectural features. There is ample evidence that the fusogenic function of representative fusion proteins is influenced by the primary structure of the single transmembrane domain (TMD) and the region linking it to the soluble assembly domains. Here, we used comprehensive non-redundant datasets to examine potential over- and underrepresentation of amino acid types in the TMDs and flanking regions relative to control proteins that share similar biosynthetic origins. Our results reveal conserved overall and/or site-specific enrichment of beta-branched residues and Gly within the TMDs, underrepresentation of Gly and Pro in regions flanking the TMD N-terminus, and overrepresentation of the same residue types in C-terminal flanks of SNAREs and viral fusion proteins. Furthermore, the basic Lys and Arg are enriched within SNARE N-terminal flanking regions. These results suggest evolutionary conservation of key structural features of fusion proteins and are discussed in light of experimental findings that link these features to the fusogenic function of these proteins. PMID- 21633972 TI - Mechanistic role of NS4A and substrate in the activation of HCV NS3 protease. AB - Hepatitis C virus (HCV) NS3 protease is the key enzyme for its maturation. Three hypotheses have been advanced in the literature to demonstrate the mechanism of the activation of the HCV NS3 protease. A virus-encoded protein NS4A and substrate are proposed to be involved in the activation of the HCV NS3 protease. However, the three hypotheses are not completely consistent with one another. Multiple molecular dynamics simulations were performed on various NS3 protease systems: free NS3 protease, NS3/4A, NS3/inhibitor, and NS3/4A/inhibitor complexes, to further unravel the mechanism of the activation of the NS3 protease. Simulation results suggest that the binding of NS4A induces a classic serine protease conformation of the catalytic triad of the NS3 protease. NS4A rearranges the secondary structure of both the N-terminus and catalytic site of the NS3 protease, reduces the mobility of the global structure of the NS3 protease, especially the catalytic site, and provides a rigid and tight structure, except for the S1 pocket, for the binding and hydrolysis of substrates. The binding of substrate also contributes to the activation of the NS3 protease by an induced-fit of the classic serine protease catalytic triad. However, the global structure of the NS3 protease is still loose and highly flexible without stable secondary structural elements, such as helix alpha0 at the N-terminus and helix alpha1 and beta-sheet E1-F1 at the catalytic site. The structure of the NS3 protease without NS4A is not suitable for the binding and hydrolysis of substrates. PMID- 21633973 TI - A machine learning approach for the prediction of protein surface loop flexibility. AB - Proteins often undergo conformational changes when binding to each other. A major fraction of backbone conformational changes involves motion on the protein surface, particularly in loops. Accounting for the motion of protein surface loops represents a challenge for protein-protein docking algorithms. A first step in addressing this challenge is to distinguish protein surface loops that are likely to undergo backbone conformational changes upon protein-protein binding (mobile loops) from those that are not (stationary loops). In this study, we developed a machine learning strategy based on support vector machines (SVMs). Our SVM uses three features of loop residues in the unbound protein structures Ramachandran angles, crystallographic B-factors, and relative accessible surface area-to distinguish mobile loops from stationary ones. This method yields an average prediction accuracy of 75.3% compared with a random prediction accuracy of 50%, and an average of 0.79 area under the receiver operating characteristic (ROC) curve using cross-validation. Testing the method on an independent dataset, we obtained a prediction accuracy of 70.5%. Finally, we applied the method to 11 complexes that involve members from the Ras superfamily and achieved prediction accuracy of 92.8% for the Ras superfamily proteins and 74.4% for their binding partners. PMID- 21633974 TI - Chronically saturating levels of endogenous glycine disrupt glutamatergic neurotransmission and enhance synaptogenesis in the CA1 region of mouse hippocampus. AB - Glycine serves a dual role in neurotransmission. It is the primary inhibitory neurotransmitter in the spinal cord and brain stem and is also an obligatory coagonist at the excitatory glutamate, N-methyl-D-aspartate receptor (NMDAR). Therefore, the postsynaptic action of glycine should be strongly regulated to maintain a balance between its inhibitory and excitatory inputs. The glycine concentration at the synapse is tightly regulated by two types of glycine transporters, GlyT1 and GlyT2, located on nerve terminals or astrocytes. Genetic studies demonstrated that homozygous (GlyT1-/-) newborn mice display severe sensorimotor deficits characterized by lethargy, hypotonia, and hyporesponsivity to tactile stimuli and ultimately die in their first postnatal day. These symptoms are similar to those associated with the human disease glycine encephalopathy in which there is a high level of glycine in cerebrospinal fluid of affected individuals. The purpose of this investigation is to determine the impact of chronically high concentrations of endogenous glycine on glutamatergic neurotransmission during postnatal development using an in vivo mouse model (GlyT1+/-). The results of our study indicate the following; that compared with wild-type mice, CA1 pyramidal neurons from mutants display significant disruptions in hippocampal glutamatergic neurotransmission, as suggested by a faster kinetic of NMDAR excitatory postsynaptic currents, a lower reduction of the amplitude of NMDAR excitatory postsynaptic currents by ifenprodil, no difference in protein expression for NR2A and NR2B but a higher protein expression for PSD-95, an increase in their number of synapses and finally, enhanced neuronal excitability. PMID- 21633975 TI - Alzheimer's disease: beta-amyloid plaque formation in human brain. AB - Although the precise cause of Alzheimer's disease is not known, the beta-amyloid peptide chains of 40-42 amino acids are suspected to contribute to the disease. The beta-amyloid precursor protein is found on many types of cell membranes, and the action of secretases (beta and gamma) on this precursor protein normally releases the beta-amyloids at a high rate into the plasma and the cerebrospinal fluid. However, the concentrations of the beta-amyloids in the plasma and the spinal fluid vary considerably between laboratories. The beta-amyloids adsorb in the nanomolar concentration range to receptors on neuronal and glial cells. The beta-amyloids are internalized, become folded in the beta-folded or beta-pleated shape, and then stack on each other to form long fibrils and aggregates known as plaques. The beta-amyloids likely act as monomers, dimers, or multimers on cell membranes to interfere with neurotransmission and memory before the plaques build up. Treatment strategies include inhibitors of beta- and gamma-secretase, as well as drugs and physiological compounds to prevent aggregation of the amyloids. Several immune approaches and a cholesterol-lowering strategy are also being tested to remove the beta-amyloids. PMID- 21633976 TI - Identifying the serotonin transporter signal in Western blot studies of the neurotoxic potential of MDMA and related drugs. AB - A number of published studies have questioned the serotonin neurotoxic potential of 3,4-methylenedioxymethamphetamine (MDMA, "ecstasy") and related drugs (fenfluramine, p-chloroamphetamine) based upon results from Western blot studies using a custom synthesized serotonin transporter (SERT) antibody that found no reduction in the abundance of a 50kDa protein after substituted amphetamine treatment. The purpose of this study was to collect Western blot data using the same SERT antibody used in those studies, but with positive and negative controls to identify the SERT protein signal. A 63-68 kDa band that had the regional distribution expected of rat brain SERT, was decreased by 5,7-DHT, and was absent in SERT KO animals was identified as the SERT protein. Significant, lasting decreases in the abundance of the 63-68 kDa band were evident in the rat brain after treatment with MDMA and related drugs (FEN, PCA). Thus, when the band corresponding to the SERT protein is identified in Western blots through the use of positive and negative controls, reduced abundance of the SERT protein can be readily demonstrated after substituted amphetamine treatment. These data provide further evidence of lasting loss of the SERT protein after exposure to MDMA and other substituted amphetamines. PMID- 21633977 TI - Bioactive natural products and chirality. AB - Mori's synthetic works on bioactive natural products in general and pheromones in particular started about 40 years ago to establish their absolute configurations and also to clarify their stereochemistry-bioactivity relationships. Results indicate that bioactive natural products are not always enantiomerically pure, and the stereochemistry-bioactivity relationships are not simple but complicated. For example, neither (R)- nor (S)-sulcatol, the aggregation pheromone of an ambrosia beetle, is behaviorally bioactive, whereas their mixture is active. In the case of olean, the sex pheromone of the olive fruit fly, its (R)-isomer is active against the males and the (S)-isomer is active against the females. Recent synthesis of two new insect pheromones is discussed to illustrate the modern methods in enantioselective synthesis. PMID- 21633978 TI - Hydride mobility in trinuclear sulfido clusters with the core [Rh3(MU-H)(MU3 S)2]: molecular models for hydrogen migration on metal sulfide hydrotreating catalysts. AB - The treatment of [{Rh(MU-SH){P(OPh)(3)}(2)}(2)] with [{M(MU-Cl)(diolef)}(2)] (diolef=diolefin) in the presence of NEt(3) affords the hydrido-sulfido clusters [Rh(3)(MU-H)(MU(3)-S)(2)(diolef){P(OPh)(3)}(4)] (diolef=1,5-cyclooctadiene (cod) for 1, 2,5-norbornadiene (nbd) for 2, and tetrafluorobenzo[5,6]bicyclo[2.2.2]octa 2,5,7-triene (tfb) for 3) and [Rh(2)Ir(MU-H)(MU(3)-S)(2)(cod){P(OPh)(3)}(4)] (4). Cluster 1 can be also obtained by treating [{Rh(MU-SH){P(OPh)(3)}(2)}(2)] with [{Rh(MU-OMe)(cod)}(2)], although the main product of the reaction with [{Ir(MU OMe)(cod)}(2)] was [RhIr(2)(MU-H)(MU(3)-S)(2)(cod)(2){P(OPh)(3)}(2)] (5). The molecular structures of clusters 1 and 4 have been determined by X-ray diffraction methods. The deprotonation of a hydrosulfido ligand in [{Rh(MU SH)(CO)(PPh(3))}(2)] by [M(acac)(diolef)] (acac=acetylacetonate) results in the formation of hydrido-sulfido clusters [Rh(3)(MU-H)(MU(3)-S)(2)(CO)(2) (diolef)(PPh(3))(2)] (diolef=cod for 6, nbd for 7) and [Rh(2)Ir(MU-H)(MU(3) S)(2)(CO)(2)(cod)(PPh(3))(2)] (8). Clusters 1-3 and 5 exist in solution as two interconverting isomers with the bridging hydride ligand at different edges. Cluster 8 exists as three isomers that arise from the disposition of the PPh(3) ligands in the cluster (cis and trans) and the location of the hydride ligand. The dynamic behaviour of clusters with bulky triphenylphosphite ligands, which involves hydrogen migration from rhodium to sulfur with a switch from hydride to proton character, is significant to understand hydrogen diffusion on the surface of metal sulfide hydrotreating catalysts. PMID- 21633979 TI - Molecular-glue-triggered DNA assembly to form a robust and photoresponsive nano network. AB - A robust and photoresponsive DNA network has been designed and constructed from branched DNA and molecular glue. The molecular glue is photoswitchable and can specifically bind to G-G mismatched double-stranded DNA. The assembly process can be reversibly controlled by manipulating the wavelength of light. The approach is flexible, allowing tuning of the size, morphology as well as the cavity of the network by variation of the molar ratio and the isotropic/anisotropic character of the branched building blocks. The assembled architectures are versatile and heat tolerant. These properties should allow the use of the network in further applications. PMID- 21633980 TI - Two Cu21 clusters with pseudo-D3 symmetry derived from the D-saccharate pentaanion, C6H5O8(5-). AB - The combination of Cu(NO(3))(2), potassium hydrogen saccharate (KC(6)H(9)O(8)) and 1,10-phenanthroline (phen) yields a pair of chiral cluster compounds, each with composition Cu(21)(C(6)H(5)O(8))(6)(phen)(12)(NO(3))(12) ?solvate. One of the compounds forms as orthorhombic crystals, while the other forms cubic crystals. Each of the clusters has D(3) or approximate D(3) symmetry, but the arrangement of the saccharate ion in the clusters is quite different in the two cases. The clusters in the cubic form interact with neighbouring clusters through face-to-face pi interactions involving the phen ligands, an association that leads to the generation of very large solvent-filled spaces in the crystal structure. In contrast the clusters in the orthorhombic form are much more densely packed. At the centre of each cluster that crystallises in the orthorhombic form is a nitrate anion that binds to six Cu(II) centres. ESI mass spectral studies indicate that the Cu(21) clusters exist in solution. Solid-state magnetic studies of the cubic form of Cu(21) show that antiferromagnetic coupling occurs to leave a non-zero-spin ground state, and comparisons are made to the magnetic data for other large Cu(II) clusters. PMID- 21633981 TI - ortho-Magnesiation of boron-substituted benzenes by using (TMP)2 Mg. PMID- 21633982 TI - Purple rice (Oryza sativa L.) extract and its constituents inhibit VEGF-induced angiogenesis. AB - The study evaluated the protective effects of purple rice (Oryza sativa L.) bran extract (PRE) and its constituents, cyanidin and peonidin, against angiogenesis induced by vascular endothelial growth factor (VEGF). The effects of VEGF and PRE were examined by in vitro tube formation assays and following 14-day co-culture of human umbilical vein endothelial cells (HUVECs) and fibroblasts. The antiangiogenic mechanism of PRE was evaluated by VEGF-induced proliferation and migration of HUVECs and/or human retinal microvascular endothelial cells (HRMECs) and phosphorylation of extracellular signal-regulated kinase (ERK) and p38. The PRE significantly suppressed VEGF-induced tube formation, proliferation and migration in HUVECs and HRMECs as well as phosphorylation of ERK and p38. Cyanidin and peonidin also suppressed the proliferation and migration induced by VEGF. These findings indicate that PRE and anthocyanidins suppress VEGF-induced angiogenesis by inhibiting proliferation and migration and suggest that the inhibition of phosphorylated-ERK and -p38 may be involved in the underlying mechanism. PMID- 21633983 TI - Apo raver1 structure reveals distinct RRM domain orientations. AB - Raver1 is a multifunctional protein that modulates both alternative splicing and focal adhesion assembly by binding to the nucleoplasmic splicing repressor polypyrimidine tract protein (PTB) or to the cytoskeletal proteins vinculin and alpha-actinin. The amino-terminal region of raver1 has three RNA recognition motif (RRM1, RRM2, and RRM3) domains, and RRM1 interacts with the vinculin tail (Vt) domain and vinculin mRNA. We previously determined the crystal structure of the raver1 RRM1-3 domains in complex with Vt at 2.75 A resolution. Here, we report crystal structure of the unbound raver1 RRM1-3 domains at 2 A resolution. The apo structure reveals that a bound sulfate ion disrupts an electrostatic interaction between the RRM1 and RRM2 domains, triggering a large relative domain movement of over 30 degrees . Superposition with other RNA-bound RRM structures places the sulfate ion near the superposed RNA phosphate group suggesting that this is the raver1 RNA binding site. While several single and some tandem RRM domain structures have been described, to the best of our knowledge, this is the second report of a three-tandem RRM domain structure. PMID- 21633984 TI - Nuclear magnetic resonance spectroscopy with the stringent substrate rhodanese bound to the single-ring variant SR1 of the E. coli chaperonin GroEL. AB - Nuclear magnetic resonance (NMR) observation of the uniformly (2) H,(15) N labeled stringent 33-kDa substrate protein rhodanese in a productive complex with the uniformly (14) N-labeled 400 kDa single-ring version of the E. coli chaperonin GroEL, SR1, was achieved with the use of transverse relaxation optimized spectroscopy, cross-correlated relaxation-induced polarization transfer, and cross-correlated relaxation-enhanced polarization transfer. To characterize the NMR-observable parts of the bound rhodanese, coherence buildup rates by different magnetization transfer mechanisms were measured, and effects of covalent crosslinking of the rhodanese to the apical binding surface of SR1 were investigated. The results indicate that the NMR-observable parts of the SR1 bound rhodanese are involved in intracomplex rate processes, which are not related to binding and release of the substrate protein from the SR1 binding surface. Rather, they correspond to mobility of the stably bound substrate, which thus appears to include flexibly disordered polypeptide segments devoid of long lived secondary structures or tertiary folds, as was previously observed also with the smaller substrate human dihydrofolate reductase. PMID- 21633986 TI - High resolution crystal structures of triosephosphate isomerase complexed with its suicide inhibitors: the conformational flexibility of the catalytic glutamate in its closed, liganded active site. AB - The key residue of the active site of triosephosphate isomerase (TIM) is the catalytic glutamate, which is proposed to be important (i) as a catalytic base, for initiating the reaction, as well as (ii) for the subsequent proton shuttling steps. The structural properties of this glutamate in the liganded complex have been investigated by studying the high resolution crystal structures of typanosomal TIM, complexed with three suicide inhibitors: (S)-glycidol phosphate ((S)-GOP, at 0.99 A resolution), (R)-glycidol phosphate, ((R)-GOP, at 1.08 A resolution), and bromohydroxyacetone phosphate (BHAP, at 1.97 A resolution). The structures show that in the (S)-GOP active site this catalytic glutamate is in the well characterized, competent conformation. However, an unusual side chain conformation is observed in the (R)-GOP and BHAP complexes. In addition, Glu97, salt bridged to the catalytic lysine in the competent active site, adopts an unusual side chain conformation in these two latter complexes. The higher chemical reactivity of (S)-GOP compared with (R)-GOP, as known from solution studies, can be understood: the structures indicate that in the case of (S)-GOP, Glu167 can attack the terminal carbon of the epoxide in a stereoelectronically favored, nearly linear O?C?O arrangement, but this is not possible for the (R) GOP isomer. These structures confirm the previously proposed conformational flexibility of the catalytic glutamate in its closed, liganded state. The importance of this conformational flexibility for the proton shuttling steps in the TIM catalytic cycle, which is apparently achieved by a sliding motion of the side chain carboxylate group above the enediolate plane, is also discussed. PMID- 21633987 TI - Crystal structure and enzymatic characterization of thymidylate synthase X from Helicobacter pylori strain SS1. AB - Thymidylate synthase X (ThyX) catalyzes the methylation of dUMP to form dTMP in bacterial life cycle and is regarded as a promising target for antibiotics discovery. Helicobacter pylori is a human pathogen associated with a number of human diseases. Here, we cloned and purified the ThyX enzyme from H. pylori SS1 strain (HpThyX). The recombinant HpThyX was discovered to exhibit the maximum activity at pH 8.5, and K(m) values of the two substrates dUMP and CH(2) H(4) folate were determined to be 15.3 +/- 1.25 MUM and 0.35 +/- 0.18 mM, respectively. The analyzed crystal structure of HpThyX with the cofactor FAD and the substrate dUMP (at 2.31 A) revealed that the enzyme was a tetramer bound to four dUMP and four FAD molecules. Different from the catalytic feature of the classical thymidylate synthase (ThyA), N5 atom of the FAD functioned as a nucleophile in the catalytic reaction instead of Ser84 and Ser85 residues. Our current work is expected to help better understand the structural and enzymatic features of HpThyX thus further providing valuable information for anti-H. pylori inhibitor discovery. PMID- 21633985 TI - The N-end rule pathway and regulation by proteolysis. AB - The N-end rule relates the regulation of the in vivo half-life of a protein to the identity of its N-terminal residue. Degradation signals (degrons) that are targeted by the N-end rule pathway include a set called N-degrons. The main determinant of an N-degron is a destabilizing N-terminal residue of a protein. In eukaryotes, the N-end rule pathway is a part of the ubiquitin system and consists of two branches, the Ac/N-end rule and the Arg/N-end rule pathways. The Ac/N-end rule pathway targets proteins containing N(alpha) -terminally acetylated (Nt acetylated) residues. The Arg/N-end rule pathway recognizes unacetylated N terminal residues and involves N-terminal arginylation. Together, these branches target for degradation a majority of cellular proteins. For example, more than 80% of human proteins are cotranslationally Nt-acetylated. Thus most proteins harbor a specific degradation signal, termed (Ac)N-degron, from the moment of their birth. Specific N-end rule pathways are also present in prokaryotes and in mitochondria. Enzymes that produce N-degrons include methionine-aminopeptidases, caspases, calpains, Nt-acetylases, Nt-amidases, arginyl-transferases and leucyl transferases. Regulated degradation of specific proteins by the N-end rule pathway mediates a legion of physiological functions, including the sensing of heme, oxygen, and nitric oxide; selective elimination of misfolded proteins; the regulation of DNA repair, segregation and condensation; the signaling by G proteins; the regulation of peptide import, fat metabolism, viral and bacterial infections, apoptosis, meiosis, spermatogenesis, neurogenesis, and cardiovascular development; and the functioning of adult organs, including the pancreas and the brain. Discovered 25 years ago, this pathway continues to be a fount of biological insights. PMID- 21633988 TI - Reprogramming chaperone pathways to improve membrane protein expression in Escherichia coli. AB - Because membrane proteins are difficult to express, our understanding of their structure and function is lagging. In Escherichia coli, alpha-helical membrane protein biogenesis usually involves binding of a nascent transmembrane segment (TMS) by the signal recognition particle (SRP), delivery of the SRP-ribosome nascent chain complexes (RNC) to FtsY, a protein that serves as SRP receptor and docks to the SecYEG translocon, cotranslational insertion of the growing chain into the translocon, and lateral transfer, packing and folding of TMS in the lipid bilayer in a process that may involve chaperone YidC. Here, we explored the feasibility of reprogramming this pathway to improve the production of recombinant membrane proteins in exponentially growing E. coli with a focus on: (i) eliminating competition between SRP and chaperone trigger factor (TF) at the ribosome through gene deletion; (ii) improving RNC delivery to the inner membrane via SRP overexpression; and (iii) promoting substrate insertion and folding in the lipid bilayer by increasing YidC levels. Using a bitopic histidine kinase and two heptahelical rhodopsins as model systems, we show that the use of TF deficient cells improves the yields of membrane-integrated material threefold to sevenfold relative to the wild type, and that whereas YidC coexpression is beneficial to the production of polytopic proteins, higher levels of SRP have the opposite effect. The implications of our results on the interplay of TF, SRP, YidC, and SecYEG in membrane protein biogenesis are discussed. PMID- 21633989 TI - A matching method for improving covariate balance in cost-effectiveness analyses. AB - In cost-effectiveness analyses (CEA) that use randomized controlled trials (RCTs), covariates of prognostic importance may be imbalanced and warrant adjustment. In CEA that use non-randomized studies (NRS), the selection on observables assumption must hold for regression and matching methods to be unbiased. Even in restricted circumstances when this assumption is plausible, a key concern is how to adjust for imbalances in observed confounders. If the propensity score is misspecified, the covariates in the matched sample will be imbalanced, which can lead to conditional bias. To address covariate imbalance in CEA based on RCTs and NRS, this paper considers Genetic Matching. This matching method uses a search algorithm to directly maximize covariate balance. We compare Genetic and propensity score matching in Monte Carlo simulations and two case studies, CEA of pulmonary artery catheterization, based on an RCT and an NRS. The simulations show that Genetic Matching reduces the conditional bias and root mean squared error compared with propensity score matching. Genetic Matching achieves better covariate balance than the unadjusted analyses of the RCT data. In the NRS, Genetic Matching improves on the balance obtained from propensity score matching and gives substantively different estimates of incremental cost effectiveness. We conclude that Genetic Matching can improve balance on measured covariates in CEA that use RCTs and NRS, but with NRS, this will be insufficient to reduce bias; the selection on observables assumption must also hold. PMID- 21633990 TI - Racemase activity of B. cepacia lipase leads to dual-function asymmetric dynamic kinetic resolution of alpha-aminonitriles. PMID- 21633991 TI - Activation of thiols at a silver nanoparticle surface. PMID- 21633992 TI - A facile pathway to enantiomerically enriched 3-hydroxy-2-oxindoles: asymmetric intramolecular arylation of alpha-keto amides catalyzed by a palladium DifluorPhos complex. PMID- 21633994 TI - A uniform bimetallic rhodium/iron nanoparticle catalyst for the hydrogenation of olefins and nitroarenes. PMID- 21633995 TI - Characterization of a sugar-O-methyltransferase TiaS5 affords new Tiacumicin analogues with improved antibacterial properties and reveals substrate promiscuity. AB - The 18-membered macrocyclic glycoside tiacumicin B, an RNA polymerase inhibitor, is of great therapeutic significance in treating Clostridium difficile infections. The recent characterization of the tiacumicin B biosynthetic gene cluster from Dactylosporangium aurantiacum subsp. hamdenensis NRRL 18085 revealed the functions of two glycosyltransferases, a C-methyltransferase, an acyltransferase, two cytochrome P450s, and a tailoring dihalogenase in tiacumicin biosynthesis. Here we report the genetic confirmation and biochemical characterization of TiaS5 as a sugar-O-methyltransferase, requisite for tiacumicin B biosynthesis. The tiaS5-inactivation mutant is capable of producing 14 tiacumicin analogues (11 of which are new), all lacking the 2'-O-methyl group on the internal rhamnose moiety. Notably, two tiacumicin analogues exhibit improved antibacterial properties. We have also biochemically verified TiaS5 as an S-adenosyl-L-methionine-dependent O-methyltransferase, requiring divalent metal ions for activity. Substrate probing revealed TiaS5 to be a promiscuous enzyme, recognizing 12 tiacumicin analogues. These findings unequivocally establish that TiaS5 functions as a 2'-O-methyltransferase and provide direct biochemical evidence that TiaS5-catalyzed methylation is a tailoring step after glycosyl coupling in tiacumicin B biosynthesis. PMID- 21633996 TI - C5-DNA methyltransferase inhibitors: from screening to effects on zebrafish embryo development. AB - DNA methylation is involved in the regulation of gene expression and plays an important role in normal developmental processes and diseases, such as cancer. DNA methyltransferases are the enzymes responsible for DNA methylation on the position 5 of cytidine in a CpG context. In order to identify and characterize novel inhibitors of these enzymes, we developed a fluorescence-based throughput screening by using a short DNA duplex immobilized on 96-well plates. We have screened 114 flavones and flavanones for the inhibition of the murine catalytic Dnmt3a/3L complex and found 36 hits with IC(50) values in the lower micromolar and high nanomolar ranges. The assay, together with inhibition tests on two other methyltransferases, structure-activity relationships and docking studies, gave insights on the mechanism of inhibition. Finally, two derivatives effected zebrafish embryo development, and induced a global demethylation of the genome, at doses lower than the control drug, 5-azacytidine. PMID- 21633997 TI - Influence of the anchor group on charge transport through single-molecule junctions. PMID- 21633998 TI - Alternating-current measurements in scanning electrochemical microscopy, part 1: principle and theory. AB - The development of the scanning electrochemical microscope in ac mode is presented from both experimental and theoretical point of views. The experiments are performed with the ferri/ferrocyanide redox mediator as model system. Based on analysis of the frequency-dependent collection efficiency, diffusion between the probe and the substrate is investigated, and analysis of time constants allows evaluation of the size of the sensing area under investigation. The experimental results are in good agreement with numerical simulations. PMID- 21633999 TI - Bronsted acidity and the medium: fundamentals with a focus on ionic liquids. AB - Fundamental aspects of Bronsted acidity in ionic liquid systems, in relation to those of simple protic molecules in the gas phase, pure protic molecules in the condensed phase and solutions of protic molecules in molecular systems, are presented. The variety of acidities possible, beyond those observed in aqueous systems, is emphasised and discussed in terms of differences of solvent levelling, ionisation, dissociation, homo-/hetero-conjugate ion speciation and the stabilisation of proton-transfer products from solvent to solvent. It is argued that data regarding aqueous systems do not necessarily explain acid/base behaviour in other liquids satisfactorily. Methods of measuring acidity are reviewed, particularly by spectrophotometry and electrochemistry and recommendations proffered for estimating speciation and acidity of ionic liquids of various complexities. PMID- 21634000 TI - Therapeutic foot health education for patients with rheumatoid arthritis: a narrative review. AB - PURPOSE: Foot health interventions such as foot orthoses for people with rheumatoid arthritis (RA) reduce pain, improve function and improve overall quality of life. Additionally, patient education (PE) is considered essential in achieving good outcomes with interventions such as foot orthoses, footwear and self-care. The aim of this literature review was to identify evidence in relation to the content, use and delivery of PE in the management of RA foot problems. METHODS: An electronic search of the following databases was performed: PubMed, CINAHL, AMED, Medline and the Cochrane Library, between March 2000 and March 2010. In order to be included, studies had to be published in English, involve adults (>18 years) with RA, and assist in answering the research question. No publications regarding PE for the management of foot health-related problems in RA were found. However, other key terms emerged that embraced PE for people with RA and informed a further search. Thirty-two papers met the inclusion criteria and were reviewed with regard to the subject area, content of the paper, methodological issues and their key findings. RESULTS: The present review provides evidence for the effectiveness of PE for people with RA delivered via a staged approach, with the content and timing of education provision being driven by the needs of the patient. CONCLUSIONS: The effect of PE delivered from a podiatric context needs to be explored, and the nature and requirements of PE for individuals with RA-related foot problems from a patient and practitioner perspective requires investigation. Alternative and innovative ways of providing PE and, potentially, self-management need to be investigated and defined. PMID- 21634001 TI - Novel blue-emitting SrMg2Al16O27:Eu2+ phosphor for solid-state lighting. AB - Eu(2+)-activated SrMg(2)Al(16)O(27) novel phosphor was synthesized by a combustion method (550 degrees C furnace). The prepared phosphor was first characterized by X-ray diffraction (XRD) for confirmation of phase purity. SEM analysis showed the morphology of the phosphor. The photoluminescence characteristics showed broad-band excitation at 324 nm, which was monitored at 465 nm emission wavelength. The SrMg(2)Al(16)O(27):Eu(2+) phosphor shows broad blue emission centred at 465 nm, emitting a blue light corresponding to 4f(6) 5d(1) -> 4f(7) transition. Here we report the photoluminescence characteristics of the prepared phosphor and compare it with commercial BAM:Eu(2+) phosphor. PMID- 21634003 TI - Generation of the tamoxifen-inducible DBH-Cre transgenic mouse line DBH-CT. AB - We generated transgenic mice bearing a tamoxifen-dependent Cre recombinase expressed under the control of the dopamine-beta-hydroxylase promoter. By crossing to the ROSA26 reporter mice we show that tamoxifen-induced Cre recombinase in adult mice specifically activates beta-galactosidase expression in differentiated noradrenergic neurons of the central and peripheral nervous system. Tamoxifen application in adult mice did not induce beta-galactosidase activity in parasympathetic neurons that transiently express DBH during development. Thus, this transgenic mouse line represents a valuable tool to study gene function in mature noradrenergic neurons by conditional inactivation. PMID- 21634004 TI - Hot topic in hepatitis C virus research: the type of immunosuppression does not matter. AB - KEY POINTS: 1. The natural history of recurrent hepatitis C virus (HCV) is highly variable. Old donor age is a factor that has consistently been shown to affect disease progression. 2. Overall, immunosuppression determines the progression of HCV-related disease; however, the type of immunosuppressive agent used for induction or maintenance is not a key factor. 3. Steroid boluses should be avoided; they are associated with increased viremia, fibrosis progression, and reduced survival. 4. Antiviral therapy, particularly if it is successful, is associated with improved outcomes for liver transplant recipients with HCV. 5. There are no convincing data for modifying the type of immunosuppression before antiviral therapy is started. PMID- 21634005 TI - Comparison of quantification methods illustrates reduced Pseudomonas aeruginosa activity on nanorough polyvinyl chloride. AB - Patients on mechanical ventilators for extended periods of time are faced with a high probability of developing ventilator associated pneumonia. Although this has been mostly addressed through the re-engineering of endotracheal tubes (ETTs) with antimicrobial materials, such material coatings may easily delaminate during use. However, the potential exists to apply nanotechnology to the ETT to avoid delamination but implement antibacterial properties. Selecting a protocol to evaluate in vitro material for anti-infection is difficult, partially due to the existence of conflicting reported methods of analysis. In this study, the susceptibility of conventional and nanorough polymeric materials to bacterial biofilm growth were evaluated. After creating nanorough polyvinyl chloride (PVC) ETTs, Pseudomonas aeruginosa biofilms were then grown on sample surfaces during a 24-h culture. Biofilms were then removed and assayed from sample surfaces using a variety of techniques. Comparisons between the different techniques used for biofilm removal indicated that vortexing provided adequate removal of the biofilm from sample surfaces. Most importantly, a protocol following the vortexing method of biofilm and bacteria removal provided an ~40% lower yield of colony forming units from nanorough PVC compared to conventional PVC. This suggests that Pseudomonas aeruginosa are less adherent on nanorough PVC than conventional PVC. PMID- 21634006 TI - Should the liver transplant criteria for hepatocellular carcinoma be different for deceased donation and living donation? PMID- 21634007 TI - Liver allocation and distribution: possible next steps. AB - Recent discussions about the distribution of cadaveric liver allografts for transplantation have raised many important issues. Over the past 2 years, a deliberative process including discussions, modeling, a request for information, a public forum, and a concept document has led to a greater focus on a possible path for reducing wait-list mortality. Here we describe that process, our interpretation of the feedback and responses, and possible recommendations. PMID- 21634008 TI - Six color flow cytometry detects plasma cells expressing aberrant immunophenotype in bone marrow of healthy donors. AB - BACKGROUND: Both normal and malignant plasma cell (PC) populations can be identified using modern flow cytometry (FC) technique in multiple myeloma (MM) patients. Expression of CD19 and CD56 markers is heterogenous on bone marrow PCs of healthy individuals. Little is known about immunophenotypically aberrant (CD19 /CD56+) PCs subpopulation of healthy people. METHODS: Using six color FC, we analyzed PCs in BM samples of 11 healthy donors (HD) and compared their immunophenotypic properties with clonal PC populations from MM patients. RESULTS: Both immunophenotypically normal (CD19+/CD56-) and aberrant (CD19-/CD56+) PC populations could be detected in 10 of 11 HDs' BM samples and constituted the median of 60.3% (37.3-72.3) and 9.6% (0-35.7) of BM PCs, respectively. CD19, CD56, CD38, CD45, and CD20 marker expression characteristics were of little value discriminating clonal PCs of MM patients from immunophenotypically aberrant PCs of healthy donors. CONCLUSIONS: Our findings suggest that aberrant immunophenotype is common in BM PCs of healthy people. Improvements in FC methodology to separate normal and malignant PCs remain an open area for future investigations. PMID- 21634009 TI - Interferon-lambda as a new approach for treatment of allergic asthma? PMID- 21634010 TI - Design, synthesis, ADME properties, and pharmacological activities of beta-alanyl D-histidine (D-carnosine) prodrugs with improved bioavailability. AB - beta-Alanyl-D-histidine (D-CAR, the enantiomer of the natural dipeptide carnosine) is a selective and potent sequestering agent of reactive carbonyl species (RCS) that is stable against carnosinase, but is poorly absorbed in the gastrointestinal tract. Herein we report a drug discovery approach aimed at increasing the oral bioavailability of D-CAR. In our study we designed, synthesized, and evaluated a series of novel lipophilic D-CAR prodrugs. The considered prodrugs can be divided into two categories: 1) derivatives with both terminal groups modified, in which the carboxyl terminus is always esterified while the amino terminus is protected by an amidic (N-acetyl derivatives) or a carbamate (ethyloxy or benzyloxy derivatives) function; 2) derivatives with only one terminus modified, which can be alkyl esters as well as amidic or carbamate derivatives. The prodrugs were designed considering their expected lipophilicity and their hydrolysis predicted by docking simulations on the most important human carboxylesterase (hCES1). The stability and metabolic profile of the prodrugs were studied by incubating them with rat and human serum and liver fractions. The octyl ester of D-CAR (compound 13) was chosen as a candidate for further pharmacological studies due to its rapid hydrolysis to the bioactive metabolite in vitro. Pharmacokinetic studies in rats confirmed the in vitro data and demonstrated that the oral bioavailability of D-CAR is increased 2.6-fold if given as an octyl ester relative to D-CAR. Compound 13 was then found to dose dependently (at daily doses of 3 and 30 mg kg(-1) equivalent of D-CAR) decrease the development of hypertension and dyslipidemia, to restore renal functions of Zucker fa/fa obese rats, and to inhibit the carbonylation process (AGEs and pentosidine) as well as oxidative stress (urinary 8-epi-prostaglandin F2alpha and nitrotyrosine). A plausible mechanism underlying the protective effects of 13 is RCS sequestration, as evidenced by the significant increase in the level of adduct between CAR and 4-hydroxy-trans-2-nonenal (HNE, the main RCS generated by lipid oxidation) in the urine of treated animals. PMID- 21634011 TI - Identification of xanthones as selective killers of cancer cells overexpressing the ABC transporter MRP1. AB - Multidrug-resistance protein 1 (MRP1) belongs to the ATP-binding cassette (ABC) transporter family. MRP1 mediates MDR (multidrug resistance) by causing drug efflux either by conjugation to glutathione (GSH) or by co-transport with free GSH (without covalent bonding between the drug and GSH). We recently reported that the calcium channel blocker verapamil can activate massive GSH efflux in MRP1-overexpressing cells, leading to cell death through apoptosis. However, clinical use of verapamil is hampered by its cardiotoxicity. Then, in the search for compounds that act similarly to verapamil, but without major side effects, we investigated xanthones. Herein we show that xanthones induce apoptosis among resistant cells overexpressing MRP1 similarly to the verapamil effect. Among the xanthones studied, 1,3-dihydroxy-6-methoxyxanthone was identified as the most active derivative, able to specifically kill cells transfected with human MRP1 with even greater potency than verapamil. Under the same conditions, the active xanthones have no toxic effect on control (sensitive) cells. Xanthones could therefore be considered as new potential anticancer agents for the selective treatment of MRP1-positive tumors. PMID- 21634012 TI - A self-complementary nucleoside: synthesis, solid-state structure, and fluorescence behavior. AB - A novel self-complementary nucleoside ((A)T), featuring two complementary nucleobases linked through an ethynyl group has been synthesized. The rigid aromatic nucleobases provided (A)T with a pale-blue fluorescence. Unlike most fluorescent organic molecules, nucleoside (A)T gives enhanced fluorescence in solid state. It exhibits considerably enhanced fluorescence intensity and a remarkable redshift (ca. 70 nm) in its emission maximum upon an increase in concentration or decrease in temperature as a result of the formation of aggregates stabilized through hydrogen bonding and pi-pi stacking of well organized (A) T assemblies; these interactions are also evident in the solid state structure, determined by X-ray crystallography. DFT calculations supported the preference of such aggregation processes. PMID- 21634013 TI - Fluorophore-labeled, peptide-based glycoclusters: synthesis, binding properties for lectins, and detection of carbohydrate-binding proteins in cells. AB - A facile and efficient solid-phase synthesis of linear peptide-based glycoclusters with various valences and different spatial arrangements of the sugar ligands is described. The synthetic strategy includes 1) solid-phase synthesis of fluorophore-labeled, alkyne-containing peptides, 2) coupling of azide-linked, unprotected mono-, di-, and trisaccharides to the alkyne-conjugated peptides on a solid support by click chemistry, and 3) release of the fluorophore labeled glycoclusters from the solid support. By using this methodology, 32 fluorescent glycoclusters with a valence ranging from 1 to 4 and different spatial arrangements of the sugar ligands were prepared. Lectin-binding properties of the glycoclusters were initially examined by using microarrays immobilized by various lectins. These glycoclusters were then employed to detect the cell-surface carbohydrate-binding proteins in bacteria. Finally, the uptake of glycoclusters by mammalian cells through receptor-mediated endocytosis was evaluated. The results, obtained from the in vitro and in vivo studies, indicate that the binding affinities toward immobilized and cell-surface proteins are highly dependent on the valence and spatial arrangements of the sugar ligands in glycoclusters. PMID- 21634014 TI - Catalytic production of conjugated fatty acids and oils. AB - The reactive double bonds in conjugated vegetable oils are of high interest in industry. Traditionally, conjugated vegetable oils are added to paints, varnishes, and inks to improve their drying properties, while recently there is an increased interest in their use in the production of bioplastics. Besides the industrial applications, also food manufactures are interested in conjugated vegetable oils due to their various positive health effects. While the isomer type is less important for their industrial purposes, the beneficial health effects are mainly associated with the c9,t11, t10,c12 and t9,t11 CLA isomers. The production of CLA-enriched oils as additives in functional foods thus requires a high CLA isomer selectivity. Currently, CLAs are produced by conjugation of oils high in linoleic acid, for example soybean and safflower oil, using homogeneous bases. Although high CLA productivities and very high isomer selectivities are obtained, this process faces many ecological drawbacks. Moreover, CLA-enriched oils can not be produced directly with the homogeneous bases. Literature reports describe many catalytic processes to conjugate linoleic acid, linoleic acid methyl ester, and vegetable oils rich in linoleic acid: biocatalysts, for example enzymes and cells; metal catalysts, for example homogeneous metal complexes and heterogeneous catalysts; and photocatalysts. This Review discusses state-of-the-art catalytic processes in comparison with some new catalytic production routes. For each category of catalytic process, the CLA productivities and the CLA isomer selectivity are compared. Heterogeneous catalysis seems the most attractive approach for CLA production due to its easy recovery process, provided that the competing hydrogenation reaction is limited and the CLA production rate competes with the current homogeneous base catalysis. The most important criteria to obtain high CLA productivity and isomer selectivity are (1) absence of a hydrogen donor, (2) absence of catalyst acidity, (3) high metal dispersion, and (4) highly accessible pore architecture. PMID- 21634015 TI - Toward molecular genetic dissection of neural circuits for emotional and motivational behaviors. AB - How does the brain process the emotional meaning of sensory stimuli and in turn drive behavior?Studies in the mammalian systems have identified various brain regions and neurotransmitter systems that are critical for emotional and motivational behaviors and have implicated their involvement in neuropsychiatric disorders including anxiety, depression, schizophrenia, and addiction. Despite these significant advancements, the precise neural circuitry underlying emotional and motivational behaviors remains to be understood at molecular and cellular levels. In this review, we discuss how the vertebrate model organism zebrafish can help us gain insights into the underlying circuitry. We first describe studies of several simple and relevant preference behaviors in this model organism, and then discuss approaches and technologies that can be used to uncover the development and function of neural circuits underlying these behaviors. PMID- 21634016 TI - The Xenopus retinal ganglion cell as a model neuron to study the establishment of neuronal connectivity. AB - Neurons receive inputs through their multiple branched dendrites and pass this information on to the next neuron via long axons, which branch within the target. The shape the neuron acquires is thus the key to its proper functioning in the neural circuit in which it participates. Both axons and dendrites grow in a directed fashion to their target partner neurons by responding to a large number of molecular cues in the milieu through which they extend. They then go through the process of synaptogenesis, first choosing a neuron on which to synapse, and then the appropriate subcellular location. How a neuron acquires its unique shape, establishes and modifies appropriate synaptic connectivity, and the molecular signals involved, are key questions in developmental neurobiology. Such questions of nervous system wiring are being pursued actively with a variety of different animal models and neuron types, each with its own unique advantages. Among these, the developing retinal ganglion cell (RGC) of the South African clawed frog, Xenopus laevis, has proven particularly fruitful for revealing the secrets of how axons and dendrites acquire their final morphology and connectivity. In this review, we describe how this system can be used to understand the multiple molecular events that instruct the incorporation of RGCs into the neural circuit that controls vision. PMID- 21634017 TI - Transcriptional response to foraging experience in the honey bee mushroom bodies. AB - Enriched environmental conditions induce neuroanatomical plasticity in a variety of vertebrate and invertebrate species. We explored the molecular processes associated with experience-induced plasticity, using naturally occurring foraging behavior in adult worker honey bees (Apis mellifera). In honey bees, the mushroom bodies exhibit neuroanatomical plasticity that is dependent on accumulated foraging experience. To investigate molecular processes associated with foraging experience, we performed a time-course microarray study to examine gene expression changes in the mushroom bodies as a function of days foraged. We found almost 500 genes that were regulated by duration of foraging experience. Bioinformatic analyses of these genes suggest that foraging experience is associated with multiple molecular processes in the mushroom bodies, including some that may contribute directly to neuropil growth, and others that could potentially protect the brain from the effects of aging and physiological stress. PMID- 21634018 TI - Differences in fibre type composition between human masseter and biceps muscles in young and adults reveal unique masseter fibre type growth pattern. AB - The human jaw system is different from those of other primates, carnivores, ruminants, and rodents in temporomandibular joint and muscle anatomy. In adults, jaw muscles also differ markedly from limb and trunk muscles in composition and distribution of fibre types. It can be assumed that age-related changes between young age to adulthood in terms of craniofacial growth, teeth eruption, and improvement of jaw functions are paralleled by alterations also in composition and distribution of jaw muscle fibre types. To address this question, we have examined the fibre type composition of the human masseter, a jaw closing muscle, at young age. For comparison, the young biceps brachii was examined. The results were compared with previous data for adult masseter and biceps muscles. Young masseter and biceps were similar in that type I fibres outnumbered other fibre types and were of the same diameter. However, they differed in composition of other fibre types. Young masseter contained fibre types I, IM, IIC, IIAB, IIB, and scarce IIA, with regional differences, whereas young biceps showed types I, IIA, IIAB, and few IIB. Young masseter differed from young biceps also by smaller type II fibre diameter and by containing fetal MyHC. In addition, the masseter and biceps differed in age-related changes of composition and distribution of fibre types between young age and adulthood. We conclude that the human masseter is specialized in fibre types already at young age and shows a unique fibre type growth pattern, in concordance with being a separate allotype of muscle. PMID- 21634019 TI - Effect of reduced c-Kit signaling on bone marrow adiposity. AB - c-Kit (CD117) is required for normal differentiation of osteoblasts from bone marrow stromal cells and for normal bone formation. Osteoblasts and adipocytes originate from a common progenitor cell, and a reciprocal relationship in differentiation of the two lineages is often observed. Therefore, the effects of abnormal c-kit signaling on bone marrow adiposity and adipocyte precursor pool size were evaluated in mouse strains with loss of function mutations in kit receptor or kit ligand. Additionally, to determine whether short-duration pharmacological disruption of kit signaling influences bone marrow adiposity, we administered the kit receptor antagonist gleevec (imatinib mesilate) for 1 week to middle aged (13-month-old) male rats known to have high levels of bone marrow fat. Compared to wild-type littermates, adipocytes were absent and adipocyte precursors greatly reduced in bone marrow from kit receptor-deficient Kit(W/W-nu) mice. Administration of secreted kit ligand to membrane-associated kit ligand deficient Kit(Sl/Sl-d) mice was ineffective in inducing bone marrow adipogenesis. These findings suggest that activation of kit receptor by the membrane-associated form of kit ligand is required for kit signaling to promote bone marrow adipogenesis in mice. Rats treated with gleevec had lower adipocyte density compared to age-matched controls, suggesting that kit signaling is required to maintain normal bone marrow adiposity. Taken together, our results indicate that c-Kit signaling plays an important but previously unsuspected role in regulating bone marrow adiposity. PMID- 21634020 TI - The anatomy of the dog soft palate. II. Histological evaluation of the caudal soft palate in brachycephalic breeds with grade I brachycephalic airway obstructive syndrome. AB - In brachycephalic dogs, the skull bone shortening is not paralleled by a decreased development of soft tissues. Relatively longer soft palate is one of the main factors contributing to pharyngeal narrowing during normal respiratory activity of these dog breeds, which are frequent carriers of the brachycephalic airway obstructive syndrome (BAOS), which affects most part of them during their postnatal life. No histological studies assessing the morphology and the normal tissue composition of the soft palate in brachycephalic dogs are available, neither has ever been determined whether the elongated soft palate is a primary or secondary event. Aim of this study was to describe the morphology of the caudal soft palate in brachycephalic dogs with Grade I BAOS to identify potential features possibly favoring the pathogenesis of BAOS. Specimens from brachycephalic dogs (N = 11) that underwent preventive surgery were collected from surgery, processed for histology, and examined at six transversal levels. The brachycephalic soft palates showed peculiar features such as thickened superficial epithelium, extensive oedema of the connective tissue, and mucous gland hyperplasia. Several muscular alterations were evidenced in addition. The results of this investigation add to the general knowledge of the anatomy of soft palate in the canine species and establish baseline information on the morphological basis of the soft palate thickening in brachycephalic dogs. PMID- 21634021 TI - The anatomy of the dog soft palate. I. Histological evaluation of the caudal soft palate in mesaticephalic breeds. AB - The gross anatomy and overall structure of the soft palate has been described in the average dog's head, however, no descriptive microanatomical studies of the dog soft palate are available, despite their possible utility in view of the manifold and important repercussions of this organ physiology. This is the first of two companion papers, dealing with the caudal part of the soft palate in the canine species, in mesaticephalic and brachycephalic dogs. Specimens from mesaticephalic healthy dogs (N = 8) were collected after euthanasia, processed for histology and sectioned at six transversal levels. Morphological stainings were used for a microscopic evaluation of the tissue layers composing the distal part of the soft palate in adult mesaticephalic dogs, and histochemical reactions were applied to assess mucin types within glandular tissue and to investigate the connective tissues. The organ was characteristically organized into a major deep musculo-connective axis mixed with salivary glands and covered by the mucosal lining on either the nasopharyngeal or the oral sides. The results of this investigation add to the general knowledge of the anatomy of soft palate in the canine species and establish baseline information for the parallel study on the long and thickened soft palate, which is typical of adult brachycephalic dogs. PMID- 21634022 TI - Glutamate microinjected in the posterodorsal medial amygdala induces subtle increase in the consumption of a three-choice macronutrient self-selection diet in male rats. AB - Previous studies have involved the "posterodorsal" amygdaloid area with the control of food intake and the development of obesity in rats. Within this wide region, the posterodorsal medial amygdala (MePD) has connections with specific hypothalamic nuclei that increase feeding behavior and modulate energy balance. Glutamate is the major brain excitatory neurotransmitter, remarkably enhances centrally mediated food consumption, and is abundantly found in the MePD. Here, it was studied the effects of saline (0.3 MUL) and glutamate (45 nM or 45 mM/0.3 MUL) directly microinjected in the MePD of adult male rats on the consumption of a three-choice (high-carbohydrate, high-protein, or high-lipid) macronutrient selective diet. The rat adaptation to the experimental procedures and its body weight gain were continuously evaluated. Control data for all groups and results following microinjections were obtained after a fasting protocol. Feeding behavior was evaluated during the subsequent 2-hr period of free access to the selective diets. Both doses of glutamate microinjected in the MePD did not lead to a higher percentage of animals consuming any of the different diets (P > 0.05), although glutamate 45 mM induced a higher consumption of the high carbohydrate diet when compared with presurgery control values (P < 0.01). Interestingly, present data indicate that glutamate in the male MePD induces only a subtle modification in the feeding behavior and suggest that large electrolytic lesions of the "posterodorsal" amygdaloid region might have affected other regions to alter drastically meal size consumption in rats. PMID- 21634023 TI - Virtual assessment of the endocranial morphology of the early modern European fossil calvaria from cioclovina, romania. AB - Endocasts provide evidence on size and shape characteristics, blood supply trajectories, and neurological features of the brain, allowing comparative analyses of fossil hominins crucial to our understanding of human brain evolution. Here, we assess the morphological features of the virtual endocast of the Cioclovina Upper Paleolithic calvarium, one of the earliest reliably dated European modern human fossils. Our study was conducted on a computed tomography (CT) scan of the original specimen. The endocranial profile was approximated via a semiautomatic segmentation of the CT data. Virtual reconstructions of the endocast were used for assessing the morphological features of the endocranium and for the estimation of the endocranial volume. Cioclovina exhibits a clockwise torque with a small anterior extension of the left frontal lobe over the right one and a protrusion of the right occipital lobe over the left, most likely due to the superior sagittal sinus coursing over the occipital pole. There is an obvious right predominance of the posterior drainage system. Interestingly, the area of the frontal sinus is occupied by dense bony tissue with small air cells corresponding probably to a natural bony loss in the diploe and to vascular spaces. An estimated endocranial volume of 1498.53 cc was calculated. The convolutional details of the third inferior frontal gyrus (Broca's caps) are indistinguishable from those found in modern Homo sapiens, and the left occipital lobe appears wider than the right, a possible correlate of right-handedness. Our metric analysis of endocranial measurements also aligns Cioclovina with modern humans. PMID- 21634024 TI - Anatomy comic strips. AB - Comics are powerful visual messages that convey immediate visceral meaning in ways that conventional texts often cannot. This article's authors created comic strips to teach anatomy more interestingly and effectively. Four-frame comic strips were conceptualized from a set of anatomy-related humorous stories gathered from the authors' collective imagination. The comics were drawn on paper and then recreated with digital graphics software. More than 500 comic strips have been drawn and labeled in Korean language, and some of them have been translated into English. All comic strips can be viewed on the Department of Anatomy homepage at the Ajou University School of Medicine, Suwon, Republic of Korea. The comic strips were written and drawn by experienced anatomists, and responses from viewers have generally been favorable. These anatomy comic strips, designed to help students learn the complexities of anatomy in a straightforward and humorous way, are expected to be improved further by the authors and other interested anatomists. PMID- 21634026 TI - Inhibition of myosin II triggers morphological transition and increased nuclear motility. AB - We investigate the effect of myosin II inhibition on cell shape and nuclear motility in cultures of mouse radial glia-like neural progenitor and rat glioma C6 cells. Instead of reducing nucleokinesis, the myosin II inhibitor blebbistatin provokes an elongated bipolar morphology and increased nuclear motility in both cell types. When myosin II is active, time-resolved traction force measurements indicate a pulling force between the leading edge and the nucleus of C6 cells. In the absence of myosin II activity, traction forces during nucleokinesis are diminished below the sensitivity threshold of our assay. By visualizing the centrosome position in C6 cells with GFP-centrin, we show that in the presence or absence of myosin II activity, the nucleus tends to overtake or lag behind the centrosome, respectively. We interpret these findings with the help of a simple viscoelastic model of the cytoskeleton consisting active contractile and passive compressed elements. PMID- 21634025 TI - Ciliary diffusion barrier: the gatekeeper for the primary cilium compartment. AB - The primary cilium is a cellular antenna that detects and transmits chemical and mechanical cues in the environment through receptors and downstream signal proteins enriched along the ciliary membrane. While it is known that ciliary membrane proteins enter the cilium by way of vesicular and intraflagellar transport, less is known about how ciliary membrane proteins are retained in, and how apical membrane proteins are excluded from the cilium. Here, we review evidence for a membrane diffusion barrier at the base of the primary cilium, and highlight the recent finding of a septin cytoskeleton diffusion barrier. We also discuss candidate ciliopathy genes that may be involved in formation of the barrier, and the role of a diffusion barrier as a common mechanism for compartmentalizing membranes and lipid domains. PMID- 21634027 TI - Diverse protective roles of the actin cytoskeleton during oxidative stress. AB - Actin oxidation is known to result in changes in cytoskeleton organization and dynamics. Actin oxidation is clinically relevant since it occurs in the erythrocytes of sickle cell patients and may be the direct cause of the lack of morphological plasticity observed in irreversibly sickled red blood cells (ISCs). During episodes of crisis, ISCs accumulate C284-C373 intramolecularly disulfide bonded actin, which reduces actin filament dynamics. Actin cysteines 284 and 373 (285 and 374 in yeast) are conserved, suggesting that they play an important functional role. We have been investigating the physiological roles of these cysteines using the model eukaryote Saccharomyces cerevisiae in response to oxidative stress load. During acute oxidative stress, all of the F-actin in wild type cells collapses into a few puncta that we call oxidation-induced actin bodies (OABs). In contrast, during acute oxidative stress the actin cytoskeleton in Cys-to-Ala actin mutants remains polarized longer, OABs are slower to form, and the cells recover more slowly than wild-type cells, suggesting that the OABs play a protective role. Live cell imaging revealed that OABs are large, immobile structures that contain actin-binding proteins and that can form by the fusion of actin cortical patches. We propose that actin's C285 and C374 may help to protect the cell from oxidative stress arising from normal oxidative metabolism and contribute to the cell's general adaptive response to oxidative stress. PMID- 21634028 TI - Regulation of beta-tubulin isotypes by micro-RNA 100 in MCF7 breast cancer cells. AB - Antimitotic drugs are key components of combination chemotherapy protocols for hematological and solid tumors. The taxanes (e.g., paclitaxel) bind to the beta subunit of the tubulin heterodimer and reduce microtubule dynamics, leading to cell cycle arrest in G2/M. The effectiveness of combination chemotherapy is limited by tumor resistance to drugs initially or as a cumulative effect after several cycles of treatment. Because changes in the drug receptor may be linked to drug resistance, we investigated changes in beta-tubulin isotypes in response to paclitaxel treatment in MCF7 breast cancer cells. We found that paclitaxel induced a 2-3 fold increase in mRNA for beta-tubulin IIA and III genes, TUBB2A, and TUBB3. beta-Tubulin class III protein increased; however, beta-tubulin class II protein was not detected in these cells. Paclitaxel treatment following pretreatment with actinomycin D showed that the change in beta-tubulin class III was due to increased transcription and linked to G2/M arrest. The increase in beta-tubulin IIA mRNA was due to both enhanced stability and increased transcription, unassociated with G2/M arrest. We used micro-RNA superarrays to look for changes in families of micro-RNAs that might be linked to drug-induced changes in beta-tubulin isotype mRNA and/or protein. We found a significant decrease in the tumor suppressor, miR-100, in MCF7 cells in response to paclitaxel treatment. Transfection of MCF7 cells with miR-100 significantly reduced beta-tubulin I, IIA, IIB and V mRNA and prevented paclitaxel-induced increases in beta-tubulin isotypes. This is the first report of a micro-RNA that regulates these specific beta-tubulin isotype mRNAs. PMID- 21634029 TI - Can we use portable nitric oxide analyzer in young children? AB - OBJECTIVE: Management of asthma could be improved by measuring exhaled nitric oxide (FENO). Portable hand-held FENO analyzer (NIOX MINO) is practical and small and could be used also in the primary care office. It has demonstrated good repeatability and correlation with stationary device (NIOX) in adults and school aged children, but so far there have been no reports on young children. The aim of this study was to compare conventional chemiluminescence device (NIOX) with a hand-held electrochemical device (NIOX MINO) in young children. DESIGN: Paired measurements of FENO were performed with the stationary chemiluminescence-based analyzer (NIOX) and with portable electrochemical device (NIOX MINO) in children with asthmatic symptoms and age-matched controls. RESULTS: Fifty-five children with mean (range) age of 5.7 (3.9-8.5) years were evaluated with both devices. Measurements were successful with both devices in 40 out of 57 children. NIOX MINO was more difficult to use than NIOX in this age group, success rates being 73% and 93%, respectively (P = 0.004). The reproducibility was similar and there was a close correlation between FENO measured by the two devices (r = 0.97, P < 0.001). However, Bland-Altman plot demonstrated limits of agreement that were relatively wide compared to low levels of FENO in the sample. Both devices were sensitive enough to distinguish higher FENO levels in children with asthmatic symptoms, compared to healthy controls. CONCLUSIONS: We conclude that NIOX MINO can be used as a screening tool for the assessment of airway inflammation in children from the age of 4 years, but its applicability is limited by lower measurement success rate and relatively poor accuracy and detection limit at low levels of FENO. PMID- 21634030 TI - Domestic exposure to aeroallergens in Hong Kong families with asthmatic children. AB - Indoor aeroallergen exposures increased asthma symptoms in Caucasians, but their determinants and relationship to asthma and allergy in Asians are unclear. This study investigated exposures to cat, cockroach, and Blomia tropicalis allergens in 115 Hong Kong families with asthmatic children. Patients underwent exhaled nitric oxide and spirometric measurements. Home visits were made within 2 weeks during which parents completed a standardized questionnaire. Fel d 1, Bla g 2, and Blo t 5 in dust samples collected from patients' mattresses, bedroom floors, and living room floors were measured by immunoassays. These aeroallergens were only detectable in some homes (38-55% for Fel d 1; 9-21% for Bla g 2, and 7-14% for Blo t 5). The presence of cat and/or dog was a strong determinant for Fel d 1 in all indoor sites. The timing and frequency of bedding change was associated with Bla g 2 levels, whereas the timing of bedroom floor cleaning was a consistent factor for Blo t 5 levels. Asthmatic children in families with high allergen exposure were more likely to have >=4 wheezing attacks in preceding 12 months and exercise-induced wheezing than those with normal allergen exposure (P = 0.051 and 0.030, respectively). Mattress levels of all three allergens were also associated with severity of several allergy symptoms (P = 0.025-0.005). None of these aeroallergens correlated with exhaled nitric oxide and spirometric parameters. This study identifies determinants for cat, cockroach, and B. tropicalis levels in Hong Kong families with asthmatic children. These exposures are associated with severity of allergy symptoms. PMID- 21634031 TI - Granulomatous pleuritis caused by histoplasmosis in a healthy child. AB - Pneumonia with pleuritis is a rare presentation of histoplasmosis infection. We present a 12-year old previously healthy boy in whom histoplasmosis presented with pleuritis, confirmed by detection of antigen in empyema fluid. PMID- 21634032 TI - Bilateral lung transplantation for pediatric idiopathic pulmonary arterial hypertension: a multi-center experience. AB - Many children with idiopathic pulmonary arterial hypertension (IPAH) experience disease progression despite advanced medical therapy. In these children, heart lung or bilateral lung transplantation (BLTx) remain the only therapeutic options when other treatments fail. Data on functional outcome after BLTx in children with IPAH are limited. We report a multi-center experience of BLTx for pediatric IPAH. We performed a retrospective study including 25 centers within the International Pediatric Lung Transplant Collaborative. Children with IPAH who underwent BLTx were included (1996-2006). Twenty-three children underwent BLTx for IPAH, most of whom were in WHO class III or IV level of function pre transplantation. At 6 months post-transplantation, 82% of children reported improvement in level of function to WHO class I. The median FEV(1) was 89% (12 126) of predicted at 12 months post-transplantation. Ten patients (44%) developed BOS at a median of 42 months (3-85), of whom five died at a median of 27 months (4-86) post-transplantation. Overall mortality was 4% at 3 months post transplantation. The median survival for children in this cohort was 45 months (2 123). Our data suggest that BLTx is a valuable therapeutic option for children with end-stage IPAH with outcomes comparable to that after heart-lung transplantation in children with pulmonary arterial hypertension or those patients undergoing lung transplantation for other indications. In the majority of children, a good cardiopulmonary function is possible following BLTx, making BLTx a good therapeutic option and maximizing donor organ utilization by allowing more hearts to be available for children needing cardiac transplantation. PMID- 21634033 TI - Evaluation of impulse oscillometry during bronchial challenge testing in children. AB - BACKGROUND: The impulse oscillation system (IOS) allows easy measurement of respiratory system impedance (Zrs). The aim of this retrospective study was to evaluate the accuracy of IOS parameters obtained during methacholine challenge by comparison with "the gold standard" forced expiratory volume in the first second (FEV1). METHODS: Measurements of FEV1 and resistances at 5 and 20 Hz, reactance at 5 Hz, impedance at 5 Hz and resonant frequency were performed in 227 children with suspected asthma, before and during methacholine challenge. Data were analyzed in the overall population and in three subgroups according to the final diagnosis: asthma (n = 72), chronic cough and nonspecific respiratory symptoms (n = 122), allergic rhinitis (n = 33). RESULTS: All IOS parameters changed significantly during the tests but only changes in X5 were significantly different between responders and nonresponders. Moreover, changes in IOS parameters were not correlated with changes in FEV1 apart from a weak correlation for X5. The receiver operating characteristic (ROC) curve for changes in X5 (to predict a 20% decrease in FEV1 showed a best decision level for a 50% decrease in X5 with a sensitivity of 36% and a specificity of 85%. Results were not different in the asthma group. CONCLUSION: The accuracy of measurements by IOS during methacholine bronchial challenge in children was not suitable when compared with FEV1 . It could be assumed that spirometry and IOS, while both providing indirect indices of airway patency, are exploring different mechanisms, each with its own methodological potentials and limitations. PMID- 21634034 TI - Differences in dinucleotide frequencies of thermophilic genes encoding water soluble and membrane proteins. AB - The occurrence frequencies of the dinucleotides of genes of three thermophilic and three mesophilic species from both archaea and eubacteria were investigated in this study. The genes encoding water soluble proteins were rich in the dinucleotides of purine dimers, whereas the genes encoding membrane proteins were rich in pyrimidine dimers. The dinucleotides of purine dimers are the counterparts of pyrimidine dimers in a double-stranded DNA. The purine/pyrimidine dimers were favored in the thermophiles but not in the mesophiles, based on comparisons of observed and expected frequencies. This finding is in agreement with our previous study which showed that purine/pyrimidine dimers are positive factors that increase the thermal stability of DNA. The dinucleotides AA, AG, and GA are components of the codons of charged residues of Glu, Asp, Lys, and Arg, and the dinucleotides TT, CT, and TC are components of the codons of hydrophobic residues of Leu, Ile, and Phe. This is consistent with the suitabilities of the different amino acid residues for water soluble and membrane proteins. Our analysis provides a picture of how thermophilic species produce water soluble and membrane proteins with distinctive characters: the genes encoding water soluble proteins use DNA sequences rich in purine dimers, and the genes encoding membrane proteins use DNA sequences rich in pyrimidine dimers on the opposite strand. PMID- 21634035 TI - HLA polymorphism of the Zhuang population reflects the common HLA characteristics among Zhuang-Dong language-speaking populations. AB - A study of the human leukocyte antigen (HLA) genetic characteristics in the Zhuang, the largest ethnic population in China, would provide insight into Zhuang history and give a useful tool for disease associations, transplantation, and anthropology. In the present study, we report the comprehensive HLA-A, HLA-B, HLA C, and HLA-DRB1 alleles and haplotypes in the Zhuang population of southern China for the first time. A total of 13 HLA-A, 24 HLA-B, 22 HLA-C, and 18 HLA-DRB1 were identified in 104 Zhuang individuals. The frequencies of HLA-A*11:01, A*02:07, A*24:02, A*02:03, and A*33:03 on A loci, B*15:02, B*58:01, B*46:01, and B*13:01 on B loci, C*03:04, C*08:01, C*01:02, C*03:02, and C*07:02 on C loci, and DRB1*15:01, DRB1*16:02, DRB1*14:01, DRB1*15:02, and DRB1*03:01 on the DRB1 loci were >10%. The A*33:03-C*03:02-B*58:01-DRB1*03:01 and A*02:07-C*01:02-B*46:01 DRB1*14:01 haplotypes were predominant in the Zhuang. The phylogenetic tree, as well as the analysis of haplotypes, suggested that the Zhuang are genetically similar to southern Chinese populations, especially the Zhuang-Dong language speaking populations, such as the Bouyei, Dai, and Maonan. Even though the Zhuang and southern Chinese populations shared common alleles and haplotypes, the Zhuang has maintained its unique genetic characteristics. PMID- 21634036 TI - Fine mapping and candidate gene prediction of the photoperiod and thermo sensitive genic male sterile gene pms1(t) in rice. AB - Pei'ai64S, an indica sterile variety with photoperiod and thermo-sensitive genic male sterile (PTGMS) genes, has been widely exploited for commercial seed production for "two-line" hybrid rice in China. One PTGMS gene from Pei'ai64S, pms1(t), was mapped by a strategy of bulked-extreme and recessive-class approach with simple sequence repeat (SSR) and insert and deletion (In-Del) markers. Using linkage analysis for the F(2) mapping population consisting of 320 completely male sterile individuals derived from a cross between Pei'ai64S and 93-11 (indica restorer) lines, the pms1(t) gene was delimited to the region between the RM21242 (0.2 cM) and YF11 (0.2 cM) markers on the short arm of chromosome 7. The interval containing the pms1(t) locus, which was co-segregated with RM6776, is a 101.1 kb region based on the Nipponbare rice genome. Fourteen predicted loci were found in this region by the Institute for Genomic Research (TIGR) Genomic Annotation. Based on the function of the locus LOC_Os07g12130 by bioinformatics analysis, it is predicted to encode a protein containing a Myb-like DNA-binding domain, and may process the transcript with thermosensory response. The reverse transcription polymerase chain reaction (RT-PCR) results revealed that the mRNA levels of LOC_Os07g12130 were altered in different photoperiod and temperature treatments. Thus, the LOC_Os07g12130 locus is the most likely candidate gene for pms1(t). These results may facilitate not only using the molecular marker assisted selection of PTGMS genes, but also cloning of the pms1(t) gene itself. PMID- 21634037 TI - Effects of the crude extract of Polygala tenuifolia Willd on human sperm in vitro. AB - The aim of the present study is to analyze sperm membrane changes and the spermicidal effect in treatment with the crude extract from Polygala tenuifolia Willd (PTW) in vitro. The root of PTW was extracted in distilled water. Normal human spermatozoa were used to assess the spermicidal activity (Sander-Cramer assay) of the extract from the PTW root. The hypo-osmotic swelling (HOS) test and the eosin Y (EY) staining were used to detect the integrity of sperm membrane and vitality. The sperm chromatin dispersion (SCD) test was performed to determine sperm DNA integrity. N-9 was used as a reference standard and semen added to physiological saline was used as the control. Semen samples were donated by 42 healthy fertile men. The crude extract from the root of PTW could immobilize and kill 100% spermatozoa within 20 s in vitro at the concentrations of 20.0 and 10.0 mg/ml; at the concentration of 5.0 mg/ml, spermatozoa were immobilized in (39.5+/ 3.2) s. In the groups of the crude extract from the root of PTW and N-9 solution, the rate of the normal HOS (tails swollen) and the white head (unstained) was 0%, and the rate of the abnormal HOS (tails unswollen) and red head (stained) was 100%. Sperm DNA fragmentation showed no change in exposure to the crude extract from the root of PTW and N-9 solution. The sperm revival test did not show any spermatozoa that recovered their motilities. The rapid spermicidal activity of the crude extract from the root of PTW in vitro may occur by the disruption of the sperm membrane integrity. PMID- 21634038 TI - Effect of tadalafil in chronic renal failure rabbits: relevance to erectile dysfunction. AB - It is of great importance to investigate an effective and reliable medication against chronic renal failure (CRF)-related erectile dysfunction (ED), which aims to improve patients' life qualities. The concentrations of cyclic guanosine monophosphate (cGMP) in the corpus cavernosal smooth muscle of both CRF and control rabbits were measured. The effects of various concentrations of tadalafil, papaverine, and sodium nitroprusside on the relaxation responses of corpus cavernosal smooth muscle pre-contracted with phenylephrine in CRF rabbits were observed. There was significant difference in the concentration of cGMP between CRF and control rabbits (P<0.01). Tadalafil had the greatest impacts on CRF rabbits when given the same concentration of papaverine or sodium nitroprusside and particularly significant differences were identified under the concentration levels of 10-5 and 10-4 mol/L (P<0.01). The results suggest that the cGMP concentrations of the corpus cavernosum had been greatly reduced in CRF rabbits compared with control rabbits and that tadalafil may be an ideal medication for use in the treatment of CRF-related ED. PMID- 21634039 TI - Analysis of 19-nortestosterone residue in animal tissues by ion-trap gas chromatography-tandem mass spectrometry. AB - A rapid sample treatment procedure for the gas chromatography-tandem mass spectrometry (GC-MS) determination of 19-nortestosterone (19-NT) in animal tissues has been developed. In our optimized procedures, enzymatic hydrolysis with beta-glucuronidase from Escherichia coli was performed in an acetate buffer (pH 5.2, 0.2 mol/L). Next, the homogenate was mixed with methanol and heated at 60 degrees C for 15 min, then placed in an ice-bath at -18 degrees C for 2 h. After liquid-liquid extraction with n-hexane, the analytes were subjected to a normal-phase solid phase extraction (SPE) C18 cartridge for clean-up. The dried organic extracts were derivatized with heptafluorobutyric anhydride (HFBA), and then the products were injected into GC-MS. Using electron impact mass spectrometry (EI-MS) with positive chemical ionization (PCI), four diagnostic ions (m/z 666, 453, 318, and 306) were determined. A standard calibration curve over the concentration range of 1-20 ng/g was reached, with Y=467084X-68354 (R2=0.9997) for 19-NT, and the detection limit was 0.3 ng. When applied to spiked samples collected from bovine and ovine, the recoveries ranged from 63% to 101% with relative standard deviation (RSD) between 2.7% and 8.9%. The procedure is a highly efficient, sensitive, and more economical method which offers considerable potential to resolve cases of suspected nandrolone doping in husbandry animals. PMID- 21634040 TI - Reagentless biosensor based on layer-by-layer assembly of functional multiwall carbon nanotubes and enzyme-mediator biocomposite. AB - A simple and controllable layer-by-layer (LBL) assembly method was proposed for the construction of reagentless biosensors based on electrostatic interaction between functional multiwall carbon nanotubes (MWNTs) and enzyme-mediator biocomposites. The carboxylated MWNTs were wrapped with polycations poly(allylamine hydrochloride) (PAH) and the resulting PAH-MWNTs were well dispersed and positively charged. As a water-soluble dye methylene blue (MB) could mix well with horseradish peroxidase (HRP) to form a biocompatible and negatively-charged HRP-MB biocomposite. A (PAH-MWNTs/HRP-MB)(n) bionanomultilayer was then prepared by electrostatic LBL assembly of PAH-MWNTs and HRP-MB on a polyelectrolyte precursor film-modified Au electrode. Due to the excellent biocompatibility of HRP-MB biocomposite and the uniform LBL assembly, the immobilized HRP could retain its natural bioactivity and MB could efficiently shuttle electrons between HRP and the electrode. The incorporation of MWNTs in the bionanomultilayer enhanced the surface coverage concentration of the electroactive enzyme and increased the catalytic current response of the electrode. The proposed biosensor displayed a fast response (2 s) to hydrogen peroxide with a low detection limit of 2.0*10-7 mol/L (S/N=3). This work provided a versatile platform in the further development of reagentless biosensors. PMID- 21634041 TI - Ammonium affects cell viability to inhibit root growth in Arabidopsis. AB - Ammonium (NH4+) is an important form of nitrogen nutrient for most plants, yet is also a stressor for many of them. However, the primary events of NH4+ toxicity at the cellular level are still unclear. Here, we showed that NH4+ toxicity can induce the root cell death in a temporal pattern which primarily occurs in the cells of root maturation and elongation zones, and then spreads to the cells in the meristem and root cap. The results from the NH4+-hypersensitive mutant hsn1 further confirmed our findings. Taken together, NH4+ toxicity inhibits primary root growth by inhibiting cell elongation and division and inducing root cell death. PMID- 21634042 TI - A pilot study on diagnosis of coronary artery disease using computed tomography first-pass myocardial perfusion imaging at rest. AB - BACKGROUND: Although computed tomography coronary angiography (CTCA) can identify coronary stenosis, little data exists on the ability of multislice computed tomography (MSCT) to detect myocardial perfusion defects at rest. METHODS: In 33 patients with diagnosed or suspected coronary artery disease (CAD), CTCA using retrospective electrocardiography (ECG) gating at rest and invasive coronary angiography (ICA) was performed. The 2D myocardial images were reconstructed in diastolic and systolic phases using the same raw data for CTCA. CT values of the myocardium were used as an estimate of myocardial enhancement, which were shown by color mapping. Myocardial ischemia was defined as a pattern of transient endocardial hypo-enhancement at systole and normal enhancement at diastole. The results of ICA were taken as the reference standard. RESULTS: When a diameter reduction of more than 50% in ICA was used as diagnostic criteria of CAD, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of CT first-pass myocardial perfusion imaging (MPI) at rest were 0.85, 0.67, 0.92, and 0.50 per patient, respectively, and 0.58, 0.93, 0.85, and 0.76 per vessel, respectively. CONCLUSIONS: CT first-pass MPI at rest could detect CAD patients, which could become a practical and convenient way to detect ischemia, consequently offering the ability for MSCT to act as a "one stop shop" for the diagnosis of CAD. PMID- 21634043 TI - Therapeutic efficiency of tissue-engineered human corneal endothelium transplants on rabbit primary corneal endotheliopathy. AB - To evaluate the therapeutic efficiency of tissue-engineered human corneal endothelia (TE-HCEs) on rabbit primary corneal endotheliopathy (PCEP), TE-HCEs reconstructed with monoclonal human corneal endothelial cells (mcHCECs) and modified denuded amniotic membranes (mdAMs) were transplanted into PCEP models of New Zealand white rabbits using penetrating keratoplasty. The TE-HCEs were examined using diverse techniques including slit-lamp biomicroscopy observation and pachymeter and tonometer measurements in vivo, and fluorescent microscopy, alizarin red staining, paraffin sectioning, scanning and transmission electron microscopy observations in vitro. The corneas of transplanted eyes maintained transparency for as long as 200 d without obvious edema or immune rejection. The corneal thickness of transplanted eyes decreased gradually after transplanting, reaching almost the thickness of normal eyes after 156 d, while the TE-HCE non transplanted eyes were turbid and showed obvious corneal edema. The polygonal corneal endothelial cells in the transplanted area originated from the TE-HCE transplant. An intact monolayer corneal endothelium had been reconstructed with the morphology, cell density and structure similar to those of normal rabbit corneal endothelium. In conclusion, the transplanted TE-HCE can reconstruct the integrality of corneal endothelium and restore corneal transparency and thickness in PCEP rabbits. The TE-HCE functions normally as an endothelial barrier and pump and promises to be an equivalent of HCE for clinical therapy of human PCEP. PMID- 21634044 TI - Development of a symptom expectation questionnaire for minor head injury. AB - BACKGROUND AND OBJECTIVE: Expectations and beliefs are important predictors of outcome following minor head injury. In this paper, the primary purpose is to develop a simple symptom expectation questionnaire for minor head injury for use in future research studies. METHODS: An existing database of 179 injury-naive subjects who completed a 56-item checklist of expected symptoms for minor head injury was analyzed to determine which items could correctly identify an a priori case definition of an expecter (a subject who expected at least one of these symptoms would remain chronic following minor head injury). A total of six of the 56 items were found to be discriminatory, and these were tested in additional subject groups against the original questionnaire. RESULTS: From the original database of 179 subjects completing a 56-item symptom expectation checklist, 135 expected that at least one of the 56 symptoms would be chronic following minor head injury. The 135 expecters, however, all chose at least one of six items: headache, anxious or worried, depressed, difficulty concentrating, dizziness, and neck pain. Using these six items, in two new groups of subjects, all those who endorsed one of the 56 symptoms as likely to be chronic following minor head injury (expecters) could also be identified on the 6-item checklist. CONCLUSIONS: A shortened (6-item) symptom expectation checklist of commonly reported symptoms following minor head injury (headache, anxious or worried, depressed, difficulty concentrating, dizziness, and neck pain) correctly identifies subjects who expect that at least one symptom will be chronic following minor head injury (i.e., an expecter). PMID- 21634045 TI - Is editing the roadblock to the internationalization of Chinese academic journals? AB - "China's internationalization" is in part realized through the dissemination of Chinese publications, such as academic journals. Therefore, editorial proficiency in the English language is of vital importance to the quality of the journals. This article analyzes the results of the two projects sponsored by the General Administration of Press and Publication of the People's Republic of China in 2007 and 2010 respectively, namely the "Analytical Evaluation of Language and Editing Quality of Chinese Academic Journals in English Language Issued in 2006", and the "Evaluation of Publication Quality of the China-Grown English Periodicals Issued in 2009, Sanctioned from 2005 to 2009", and proposes solutions to the problems presented therein. PMID- 21634046 TI - Octreotide. PMID- 21634047 TI - International disease monitoring, October to December, 2010. AB - African swine fever in the Caucasus and Leningrad regions of Russia. Equine infectious anaemia in several EU member states. Foot-and-mouth disease in Bulgaria--the first in an EU member state since 2007. Highly pathogenic avian influenza H5N1 in the Far East. West Nile virus in southern Europe. These are among matters discussed in the international disease monitoring report for October to December 2010, prepared by Defra's Food and Farming Group, Veterinary Science Team. PMID- 21634048 TI - Temperature and salt concentration alter base-sequence selectivity of a duplex DNA-binding protein. AB - A structural polymorphism of nucleic acids, which depends on the concentration of cations and the conditions of hydration, are strongly involved with interactions between DNA and proteins. In this paper, we report that different DNA sequences bound to hyperthermostable TATA-box-binding protein (PhoTBP) at different combinations of temperature and salt concentration in in vitro selection experiments. As a result of the interaction of-these selected DNAs with PhoTBP, characteristic changes in the numbers of water molecules and ions occurred under each condition of the selection experiment. This finding could help us to understand the solvent environment-dependent preference for base sequences in protein-DNA interactions. PMID- 21634049 TI - Volar locking plates versus Kirschner wires for distal radial fractures-A cost analysis study. PMID- 21634050 TI - The relationship between liver disease and liver fibrosis: a tangled web. PMID- 21634051 TI - Progression from isolated steatosis to steatohepatitis and fibrosis in nonalcoholic fatty liver disease. AB - In patients with nonalcoholic fatty liver disease (NAFLD) isolated steatosis is considered a benign condition with no or minimal rate of progression, in contrast to nonalcoholic steatohepatitis (NASH) which can progress to cirrhosis. We report on a series of six patients with isolated steatosis on an initial liver biopsy, and NASH on a follow-up biopsy performed five years after. All but one of the initial biopsies were longer than 15 mm. At follow-up, inflammation and ballooning were present in all patients and mild fibrosis in three. All patients had one or more features of metabolic syndrome at baseline. Progression to steatohepatitis occurred independent of aminotransferase changes. Five patients experienced an increase in one or several metabolic risk factors during follow up: body mass index, triglyceride levels, arterial hypertension and/or the HOMA index. One patient did not exhibit progression but was still exposed to metabolic risks factors at the end of follow-up. This report demonstrates that isolated steatosis is not necessarily a benign, non-progressive condition. Current recommendations for the absence of hepatic monitoring in patients with isolated steatosis are not adequate. If metabolic risk factors persist or deteriorate during follow-up and/or non-invasive markers suggest disease progression, a control liver biopsy should be considered. PMID- 21634052 TI - The genomic and physiological basis of life history variation in a butterfly metapopulation. AB - Unravelling the mechanisms underlying variation in life history traits is of fundamental importance for our understanding of adaptation by natural selection. While progress has been made in mapping fitness-related phenotypes to genotypes, mainly in a handful of model organisms, functional genomic studies of life history adaptations are still in their infancy. In particular, despite a few notable exceptions, the genomic basis of life history variation in natural populations remains poorly understood. This is especially true for the genetic underpinnings of life history phenotypes subject to diversifying selection driven by ecological dynamics in patchy environments--as opposed to adaptations involving strong directional selection owing to major environmental changes, such as latitudinal gradients, extreme climatic events or transitions from salt to freshwater. In this issue of Molecular Ecology,Wheat et al. (2011) now make a significant leap forward by applying the tools of functional genomics to dispersal-related life history variation in a butterfly metapopulation. Using a combination of microarrays, quantitative PCR and physiological measurements, the authors uncover several metabolic and endocrine factors that likely contribute to the observed life history phenotypes. By identifying molecular candidate mechanisms of fitness variation maintained by dispersal dynamics in a heterogeneous environment,they also begin to address fascinating interactions between the levels of physiology, ecology and evolution. PMID- 21634053 TI - The utility of contemporary and historical estimates of dispersal in determining response to habitat fragmentation in a tropical forest-dependent bird community. AB - It is often assumed that species which exhibit a greater propensity for dispersal are less susceptible to the impacts of habitat fragmentation; however, a growing body of literature suggests that such generalizations should be carefully evaluated as not all species appear to be equally sensitive to fragmentation. In this issue of Molecular Ecology, Callens et al. (2011) take an innovative approach to compare contemporary estimates of dispersal from an extensive mark recapture and patch occupancy data set with historical estimates derived from multilocus population genetic models for seven sympatric forest-dependent species in the Taita Hills, Africa. As has been observed for forest-dependent species from the Amazon, populations of sedentary species were more strongly differentiated and clustered when compared to those of more dispersive taxa. The most intriguing result recovered though, was that the five species with similar historical estimates of gene flow (dispersal) differed substantially in their contemporary dispersal rates, suggesting that for some species the propensity for dispersal has decreased over time. As a consequence, the authors suggest that post-fragmentation estimates of dispersal on their own may not be the best predictors of how habitat fragmentation could affect forest-dependent animal communities.This work significantly advances our understanding of the dynamics of habitat fragmentation and makes a strong case for the need to integrate data on historical processes with contemporary data. PMID- 21634054 TI - Efficacy of irinotecan in combination with 5-fluorouracil (FOLFIRI) for metastatic gastric or gastroesophageal junction adenocarcinomas (MGA) treatment. AB - OBJECTIVES: The most commonly used schedules are 5-FU in combination with CDDP with or without epirubicin (ECF) or docetaxel (TCF) in treatment of MGA patients (pts), independently of HER status. We evaluated the efficacy of FOLFIRI regimen in a large retrospective series of MGA pts. METHODS: Two hundred and twelve pts from 13 French centers were treated with at least one cycle of FOLFIRI (irinotecan 180 mg/m2 intravenous (i.v.) over 90 minutes on day 1 with folinic acid (FA) 400mg/m2 i.v. over two hours followed by 5-FU 400mg/m2 i.v. bolus then 5- FU 2400 mg/m2 continuous infusion over 46 hours on day 1, repeated every 14 days). Primary tumour sites were 120 (58%) stomach and 92 (42%) gastroesophageal junction. FOLFIRI was administered as first-line in 137 (65%) pts and as later line in 75 (35%) pts for MGA. RESULTS: There was no difference between chemonaive and not chemonaive pts treated as firstline in terms of response rate 37% (95% CI: 25-50) vs 44% (95% CI: 21-69), median PFS, 6.7 (95% CI: 5.5-9.9) vs 5.3 months (95% CI: 3.6-6.9) (P = 0.25), and OS, 13.1 (95% CI: 11.7-18.7) vs 8.8 months (95% CI: 7.3-15.6) (P = 0.19), respectively. There was no difference between pts treated as second or later-line in terms of response rate 20% (95% CI: 8-39) vs 22% (95% CI: 6-48), median PFS, four months (95% CI: 2.8-5.4) vs 3.5 months (95% CI: 2.3-4.5) (P = 0.56), and OS, 10.4 months (95% CI: 5.4-14.4) vs 5.3 months (95% CI: 3.5-11.3) (P = 0.58), respectively. The global grade 3-4 toxicities were: diarrhea 11%, vomiting 9%, neutropenia 18%, febril neutropenia 4% (one toxic death). CONCLUSIONS: This retrospective study confirms the activity and good tolerance of FOLFIRI regimen in MGA as first-line as well as later-line. PMID- 21634055 TI - Dispersal in the course of an invasion. AB - Invasive species receive attention as manifestations of global ecological change and because of the effects that they may have on other organisms. They are commonly discussed in the context of the ecological perturbations or the human activities that permitted the invasion. There is also evidence, that there is an intrinsic component to biological invasions in that evolutionary changes of the invaders themselves can facilitate or limit invasions (Lee 2002; Urban et al. 2007; Van Bocxlaer et al. 2010). Hence, teasing apart whether environmental change or changes of the organism foster invasions is an interesting field of research. Ample evidence for plants and animals documents that ecological change and human activities trigger range expansions and invasions, but questions regarding evolutionary change of invaders remain less explored although there are several reasons to believe it matters. Firstly, rapid evolutionary change is possible in time-frames relevant for contemporary biological invasions(Hendry et al. 2007). Furthermore, population genetic modelling suggests that there are circumstances where the range expansion and colonization of empty spaces in the course of an invasion can induce evolutionary change in a way that is specific to invaders: the process of repeated founding out of marginal populations in the course of a range expansion can shift allele frequencies and has been referred to as allele surfing, which not only affects neutral genetic variance, but also fitness relevant traits (Klopfstein et al. 2006; Travis et al. 2007; Burton & Travis 2008). Importantly, this process poses a null model for evolutionary inference in invasive populations. It predicts conspicuous allele frequency changes in an expanding metapopulation unless migration homogenizes the gene pool. Despite this relevance, ideas about allele surfing rely heavily on modelling although some experimental evidence comes from studies that document the segregation of genetic variants in growing plaques of bacteria (Hallatschek et al. 2007). To date, little empirical data is available that would reveal the migration processes that affect the establishment of gene pools at invasion fronts in natural systems. This aspect sets the study of Bronnenhuber et al. (2011) apart. They quantify migration behind the expansion front of an invading fish and thus provide important baseline data for the interpretation of the emerging patterns of genetic differentiation. PMID- 21634056 TI - A systematic review of adenotonsillectomy as a risk factor for childhood obesity. AB - OBJECTIVE: Tonsillectomy is the most common major surgical procedure performed in children. There is evidence that tonsillectomy is associated with weight increase and may contribute to pediatric obesity. The study aimed to review the evidence that tonsillectomy with or without adenoidectomy is a risk factor for future obesity. DATA SOURCES: Systematic literature search was performed using PubMed and Ovid. REVIEW METHODS: Systematic analysis of the literature from 1970 to 2009 on patients who underwent adenotonsillectomy (T&A) with preoperative and postoperative weight-based measurements. RESULTS: Nine studies satisfied inclusion criteria. A total of 795 children were included. Preoperative weight ranged from normal to morbid obesity. In total, 656 children had demographic information recorded, and 53.35% of the children were male. Indication for surgery was not recorded in 336 patients. In 47.7% patients, the indication recorded was sleep-disordered breathing. The first group included 3 studies involving 127 children, and body mass index (BMI) increased by 5.5% to 8.2%. The second group included 3 studies involving 419 patients; the standardized weight scores increased in 46% to 100% of patients. The third group included 3 studies with 249 patients; the corrected weight increased postoperatively in 50% to 75% of patients. Morbidly obese patients (weight 130%-260% vs peers) remained unchanged postoperatively. LIMITATIONS: Each study was designed with different definitions of overweight and a range of follow-up periods. Demographic information was limited. CONCLUSIONS: A large population of normal and overweight children undergoing T&A gained a greater than expected amount of weight postoperatively, which suggests an association between T&A and weight gain. A significant need exists for a large study with consistent outcomes measured. PMID- 21634057 TI - Nasal nitric oxide and sinonasal disease: a systematic review of published evidence. AB - OBJECTIVE: To critically and systematically review the data available on the sinonasal application of nasal nitric oxide measurement, particularly its use as a diagnostic, prognostic, or treatment effect indicator. DATA SOURCES: EMBASE 1980 to February 10, 2010; Medline 1950 to February 10, 2010; Cochrane Collaboration database; NHS Evidence Health Information Resources database. Review Methods. The databases were searched using a search strategy designed to include manuscripts relevant both to nitric oxide measurement and sinus or nasal problems. A title search was carried out on these manuscripts to select those relevant to clinical or basic science aspects of nitric oxide measurement. A subsequent abstract search selected those manuscripts concerning the application of nitric oxide measurement to sinonasal problems. The manuscripts selected were subject to a full-text review to extract data sets of nasal nitric oxide readings for different patient groups. RESULTS: Initially, 1088 manuscripts were selected. A title search found 335 manuscripts of basic scientific or clinical interest. An abstract search found 35 manuscripts directly relating to nitric oxide measurement in sinonasal disease. Full-text analysis produced 20 studies with extractable data on nasal nitric oxide levels in clearly defined patient groups. Studies did not show sufficient homogeneity to enable substantial meta-analysis of aggregated data. CONCLUSION: Current evidence shows that nasal nitric oxide is not a clinically useful measure for sinonasal disease. Although there is some evidence that sinus surgery is associated with lowered nasal nitric oxide levels, there is no evidence that this is associated with deterioration in sinus health. PMID- 21634058 TI - Veterinary Salmonella isolates in Scotland decline again in 2009. PMID- 21634059 TI - The EHRA: a short history and a long list of achievements. PMID- 21634060 TI - Do cardiologists follow their own advice? PMID- 21634063 TI - Northern Ireland Disease Surveillance Report: Northern Ireland disease surveillance, October to December 2010. PMID- 21634089 TI - [The role of VATS in the treatment of thoracic empyema]. AB - INTRODUCTION: Videothoracoscopy (VTS) and video-assisted thoracoscopic (VATS) decortication is commonly used in the treatment of early thoracic empyema. However, its conversion to thoracotomy is more frequent in chronic empyema cases. The authors evaluate the role of VTS/VATS in the pleural empyema therapy in the group of patients treated for thoracic empyema using VATS or thoracotomic decortication. PATIENTS, METHOD AND RESULTS: The retrospective study included 165 patients who underwent surgery for thoracic empyema. Pneumonia was the the commonest cause of pleural empyema (77%), while elective lung and esophageal surgery resulted in pleural empyema in 11% of the subjects, trauma in 9% and intraabdominal infection in 3% of the subjects. The subjects were diagnosed with thoracic empyema stage II or III either prior to the procedure on CT examination, or during the procedure. VTS/VATS was indicated in 52 subjects, and conversion to open decortication was required in 23 of them. Open thoracotomic decortication was performed in 113 subjects. VTS/VATS was successful in stage II disorders, in particular in the removal of pus and fibrin, with targeted chest drain insertion. VATS decortication was performed in 15 subjects with stage II or early stage III disorders. In open decortications, the procedure was extended to lung resection (8x lobectomy and 10x wedge non-anatomical resection) for residual abscess pneumonia or lung abscess. Conservatively managed postoperative complications included protracted chest drain air leak (11 patients) and fluidothorax relapses which were treated with redrainage (8 patients). Early postoperative rethoracotomy was indicated in 6 patients for hemothorax, resulting from hemocoagulation disorder in septic conditions. 4 patients were indicated for rethoracotomy for empyema relapses. During the postoperative period, 6 patients exited in the open decortication group. CONCLUSION: VTS is the method of choice in the treatment of stage II thoracic empyema, with a potential for the infectious focus removal, targeted drainage and lung reexpansion. VATS is an alternative thoracotomy method for decortication in the early stage III empyema. Decortication via thoracotomy is a standard treatment method for the management of chronic empyema. PMID- 21634090 TI - [Distant wounded glioma syndrome following stereotactic biopsy--a case review]. AB - Postoperative haemorrhage is a threatening complication of both brain tumor resection and stereotactic biopsy. The paper describes rare case report of distant wounded glioma syndrome after stereotactic biopsy of glioblastoma, when small distant bleeding was proven in the tumor nodule distant from the original site of biopsy. PMID- 21634091 TI - [Biliary complications after major liver resection]. AB - INTRODUCTION: Major liver resection involves operations where three or more liver segments are resected. The decrease of perioperative and early lethality enlarged indicator conditions, especially in patients with colorectal cancer liver metastasis. Bile leakage belongs to major postoperative complications. AIM: Authors present literary experience and retrospective analysis of patients with emphasis on biliary complications and their treatment. RESULTS: 96 patients underwent major hepatic resections between April 2004 and December 2009 at the Surgery Department of the Central Military Hospital in Prague. The average age of patients was 61 (25-84). Patients with an oncology disease dominated the set, representing 78% of all the patients. One half of the patients were formed by patients with colorectal cancer liver metastasis. Fourteen patients (14.6%) suffered from postoperative biliary complications. Non-surgical treatment was successful in nine cases. Surgical treatment was necessary in five cases. Combinations of these methods were essential in half of the patients. Two patients died (2.1%). CONCLUSION: Bile leakage after major liver resection is a quite common and serious postoperative complication. Conservative treatment (ERCP, CT - navigated drainage) is the method of choice. Surgical treatment is necessary where conservative management fails or where the size of the bile leakage is large. Multidisciplinary approach to treatment of these patients is essential. PMID- 21634092 TI - [Unified classification of the lung lymphatic system. A path to status quo]. PMID- 21634093 TI - [Benefits of the classical approach in surgery for pulmonary metastases]. AB - INTRODUCTION: Distant metastases remain a significant problem in the treatment of malignancies. Surgical management of pulmonary metastases is considered valuable from the oncological view only on condition that R0 resection can be achieved. The whole spectrum of resection procedures can be used, however most commonly, extraanatomic lung resections are employed. It has not been fully evaluated whether the same efficacy can be obtained with thoracoscopic procedures. AIM OF THE STUDY: The aim was to compare the study complication rates with literature data. The secondary aim was to evaluate the benefit of intraoperative lung palpation examination. MATERIAL AND METHODS: The authors present a retrospective study in a group of subjects operated for secondary pulmonary malignancies in the Motol Charles University 2nd Medical Faculty and Faculty Hospital Surgical Clinic, from 2003 to 2007. The authors compared the patient group's morbidity and 30-day mortality rates with literature data. Preoperative CT findings, intraoperative palpation findings and histological examination findings were assessed. RESULTS: Postoperative morbidity of the operated subjects was 16.5%, postoperative 30-day mortality was 0%. The authors compared the preoperative diagnostic data based on CT, the intraoperative findings and histological findings. During the total of 77 surgical procedures, including open and VATS procedures, the authors performed intraoperative palpation examination and detected 60 foci (24.6% out of the total removed foci) previously undetected on CT. All of the foci were of less than 5mm and in 55 cases, the foci were proved metastases. CONCLUSION: The outcome data showing low postoperative morbidity rates and nul 30-day mortality have confirmed that pulmonary metastasectomy is a safe method, a part of the complex oncological management. A surgeon's palpation finding is considered unsubstitutable in the detection of all lung foci and for necessary orientation in order to identify the safety margin in wedge resections. Therefore, the authors prefer the open or videoassissted approach to purely miniinvasive procedures. PMID- 21634094 TI - [The quality of life following laparoscopic fundoplication in correlation with preoperative symptomatology]. AB - AIM: Gastroesophageal reflux disorder (GERD) is a serious health problem in the Western world, with prevalence rates between 9 and 42%. The correct preoperative diagnostics including assessment of symptomatology is a prerequisite for the establishment of adequate therapy, including surgery. The aim of this study was to assess the quality of life in patients with GERD, based on their preoperative symptomatology. METHODS: From IX/2004 to XII/2008, a total of 237 patients underwent antireflux procedures in the Brno Faculty Hospital (FN Brno) Surgical Clinic. The patients underwent preoperative and postoperative gastroenterological examination, including endoscopy, manometry, pH-metry. The patients were asked to fill in the GIQLI (GastroIntestinal Quality of Life Index) questionnaire. The results were statistically assessed and evaluated. RESULTS: The study included a total of 178 patients. 135 subjects (76.7%) presenting with typical symptomatology and 41 subjects (23.3%) with atypical symptomatology were indicated for surgery. The mean preoperative quality of life index, based on the GIQLI questionnaire, was 101.6 points, while at 6 months after the procedure, the quality of life was evaluated with 106.9 points. There is a statistically significant difference in the quality of life between the patients with atypical and the patients with typical symptomatology, both prior to the procedure (p = 0.002), as well as after the procedure (p = 0.006), with the atypicaly symptomatology patients declaring poorer quality of life than the typical symptomatology ones. The risk of prolonged dysphagia (over 6 weeks after the procedure) is higher in the atypical symptomatology subjects, with statistical significance of p < 0.001. CONCLUSION: Laparoscopic antireflux surgery increases the quality of life independent of the preoperative symptomatology. Patients with atypical symptoms must be carefully examined and indicated with caution, considering the higher risk of prolonged postoperative dysphagia. PMID- 21634095 TI - [Program C34 CIS practicing MMX. Preliminary report]. PMID- 21634096 TI - [Impact of postoperative complications on survival of patients with pancreatic carcinoma]. AB - INTRODUCTION: In spite of radical pancreatic resection and adjuvant therapy, the overall 5-year survival rate for patients with pancreatic cancer remains poor, ranging between 5% and 15%, with a median survival of 13 to 17 months. MATERIAL AND METHODS: Between January 2000 and December 2009, the prospective study was performed to indentify the presence of postoperative complications and to find their influence on survival rate in pancreatic cancer patients. RESULTS: Between January 2000 and December 2009, the radical pancreatic resection was performed for 102 patients suffer from pancreatic cancer, at the I. Department of Surgery University Hospital in Kosice. Pancreatoduodenectomies were performed in Child Stulhofer modification in 59 patients, Whipple modification was performed in 31 patients, Waugh-Clagett in 2 patients, Traverso-Longmire in 2 patients, distal resection of pancreas was performed in 7 patients and one patient had made total pancreatectomy. The overall morbidity rate was 30.2% (31 patients) and mortality rate was 3.9% (4 patients). Specific (for pancreatic resection) complications were indentified in 26 patients, 25.3%. Non-specific complications were presented in 5 patients, 4.9%. Two intraoperative risk factors were found to be significantly associated with pancreatic leakage, small pancreatic duct size and soft texture of the remnant pancreas. The 5-years survival rate for patients with pancreatic cancer was in 5 patients, 4.9%, with median survival rate 15 months. In group of patients with postoperative complications median survival rate was 13 months, in group of patients without postoperative complications median survival time was 18 months. CONCLUSION: Long-term survival rate for pancreatic cancer patients is still low. The presence of postoperative complications had negative influence to survival rate in pancreatic cancer patients. Pancreatic leakage is the most afraid complications. Patients with a small pancreatic duct size or a soft pancreatic remnant were at high risk of pancreatic leakage. PMID- 21634097 TI - [Laparoscopic treatment of large intestinal injuries during colonoscopy]. AB - Iatrogenic colonic perforation is a rare, but a serious complication of colonoscopy. Incidence varies in between 0.03-3%. Symptoms of injury are strong abdominal pain in relation to colonoscopic examination and pneumoperitoneum on radiologic findings. Laparoscopic approach allows accurate diagnosis and definitive surgical treatment. PMID- 21634098 TI - [New options for pancreatic cancer diagnostics]. AB - INTRODUCTION: The overall 5-year survival rate for patients with pancreatic cancer remains poor, ranging between 5% and 15%, with a median survival of 13 to 17 months. Looking for the new possibilities can identify early-stage pancreatic cancer patients. MATERIAL AND METHODS: The comparative study was performed at the I. Department of Surgery University Hospital in Kosice, between group of pancreatic cancer patients, whose were operated during 1. 1. 1996-31. 12. 2000 group A and during 1. 1. 2005-31. 12. 2009 group B. The survival rate was compared in all groups of patients and in group of patients in stages of pancreatic cancer. From 2007, there were started a trial, in which lymph nodes were observed using histopathological and immunohistochemical examination. RESULTS: In group A (1. 1. 1996-31. 12. 2000) were presented 29 patients, 5 years survival was investigated in two patients (6.8%), median survival rate was 14 months. In group B (1. 1.2005-31. 12. 2009) were presented 51 patients, no patient survival 5 years, median survival rate was 14 months. Between January 2007 and December 2009 one hundred and eight pancreatic cancer patients were treated at the I. Department of Surgery University Hospital in Kosice. The radical pancreatic resection and standard lymphadenectomy were performed in 36 patients. During standard lymphadenectomy there were found 19 patients with 119 negative lymph nodes by histopathological examination. These lymph nodes were examined by immunohistological examination, and there were found 37 lymph nodes as positive, in 6 patients. CONCLUSION: Patients in earlier stage of pancreatic cancer had better survival rates than in later stage of disease. Immunohistochemical examination of histopathologically negative lymph nodes can detect positive lymph nodes and early-stage pancreatic cancer patients can be identified. PMID- 21634099 TI - [Renal cancer metastasis into common bile duct]. AB - We present rare case of clear cell renal carcinoma metastasis into common bile duct. It is a 9th case listed on the MEDLINE data base from 1966 to 2010.71-year old woman underwent right nefrectomy for renal cancer G1 in 1986. She was reoperated in 2006 for relaps in renal bed on the right side. Histological examination confirmed clear cell renal carcinoma again. At present, a new liver metastasis in S 4/5 was detected. During the operation we found first described liver metastasis and second another one in common bile duct with intraluminal growth. Before surgery the patient showed neither clinical nor laboratory marks of biliary obstruction. Even abdominal ultrasound, SONO VUE, CT and body PET were negative. We carried out choledochectomy with hepaticojejunoanastomosis sec Roux and liver metastasis was destructed using radiofrequency ablation. PMID- 21634100 TI - [Laparoscopic pancreatic resections in experimental setting and clinical practice]. AB - INTRODUCTION: Pancreatic fistula is a major postoperative complication after pancreatic resection. One of the main risk factors of developing the pancreatic fistula after distal pancreatectomy is the method employed for the management of the pancreatic remnant. AIM OF THE STUDY: The aim of the experimental part of this work was to test a novel method of management of the pancreatic remnant after distal pancreatectomy on a large laboratory animal. Furthermore, based on the experience with the experimental work to introduce the laparoscopic approach to human clinical practice. METHODS: In the experimental part of the work laparoscopic distal pancreatectomy with spleen and splenic vessels preservation was performed in ten female domestic pigs. The experimental animals were divided into two groups. In the first group the pancreas was transected using an EndoGIA Universal Stapler and in the second group, the pancreas was transected using a Ligasure device and the pancreatic remnant was reinforced with hydrogel sealant Pleuraseal. We introduced the laparoscopic distal pancreatectomy to clinical practice in the Department of Surgery in Hradec Kralove in 2009. Transection of the pancreas was performed with staplers. RESULTS: In the experimental part of the work the postoperative course was uneventful in all the animals. All animals gained weight. Only minor macroscopic and microscopic alterations of the healing process were found. Statistical differences between the groups were not significant. In the clinical part of the work we performed laparoscopic distal pancreatectomy in 6 patients. We performed two distal pancreatectomies with splenectomy, one distal pancreatectomy with splenectomy and left nephrectomy and 3 distal pancreatectomies with the spleen and splenic vessels preservation. We did not have to convert to open procedure in any of the cases. CONCLUSIONS: In the experimental part of the work we showed that the novel technique using Ligasure transection reinforced by the hydrogel sealant Pleuraseal is feasible and safe technique, which seems to be comparable with the standard transection technique using stapler. Our initial experience with laparoscopic distal pancreatectomy in the clinical practice cannot be used to compare various methods of management of the pancreatic stump or to evaluate the rate of pancreatic fistula in such small group of patients. PMID- 21634101 TI - [Laparoscopic distal pancreatectomy for neuroendocrine pancreatic tumors--initial experience]. AB - INTRODUCTION: Although the first successful laparoscopic distal pancreatectomy in Martin was recorded in 2005, after five years we have successfully established this unique surgical procedure. The aim of this paper is to present two successful laparoscopic distal pancreatectomies in patients with neuroendocrine tumors of the distal pancreas. MATERIALS AND METHODS: Laparoscopic distal pancreatic resection is currently challenging many pancreatobiliary surgeons. Its open alternative is the standard surgery for tumors in the body and tail of pancreas. Laparoscopic distal pancreatectomy meets all aspects of radical oncological resection including lymphadenectomy. Similarly to open resection is often associated with splenectomy, but brings significant benefit to the patient in the form miniinvasivity. The paper gives crucial points of surgical procedure that is still an unique surgery. RESULTS: Although the last 4 months we operated on laparoscopically only 2 patients we present at least the preliminary experience with this method as well as a rich documentation of these procedures. CONCLUSION: Laparoscopic distal pancreatectomy in the hands of an experienced laparoscopic surgeon has the chance to become an alternative to an open surgery. PMID- 21634102 TI - [Professor Dr Borivoje Gradojevic--the first university professor of orthopaedic surgery]. AB - Prof. dr Borivoje Gradojevic was one of the greatest figures of our medicine, and one of the pioneers and founders of orthopedic surgery in our country. He was the first professor of orthopedic surgery in Serbia ie. the professor of Belgrade Faculty of Medicine. Beside well education and professional skills dr Gradojevic published the numerous scientific papers and he published the first textbook of orthopedics in this country. Unfortunately, his professional career was brutally interrupted in 1945, when he was forced to retire and moved from Medical faculty, together with other professors in these times. This resulted in enormous regression--fall in academic community, and our health care system and country also. PMID- 21634103 TI - [Pathology and pathobiology of the gastric carcinoma]. AB - Recent epidemiological studies in Serbia revealed that gastric carcinoma is the third and the fifth main cause of cancer morbidity in men and women, respectively. Despite the declining incidence of gastric cancer, it remains the second most common cause of cancer-related deaths as it is worldwide. A well defined carcinogenic inflammation-metaplasia-dysplasia-cancer sequence typically precedes the development of most gastric adenocarcinomas. Alterations such as gastric mucosal atrophy and intestinal metaplasia are merely markers of increased risk, while gastric epithelial dysplasia (GED) represent a direct precursor of cancer. DNA damage and increased mucosal proliferation secondary to H pylori infection, combined with a suitable host susceptibility phenotype (eg, genetic polymorphisms in interleukin IL-1B, IL-1RN, and tumor necrosis factor a TNF-alpha genes), are important factors in this progression pathway. However, only a small minority of patients infected with H. pylori eventually develops gastric cancer, and eradication of H pylori in these patients does not seem to eliminate the risk of cancer completely. It has been shown that atrophy may be a better indicator of risk of cancer than intestinal metaplasia, and remains to be validated in routine clinical practice according to recent proposal for new quantitative methods. It is often associated with pseudopyloric gland metaplasia in the gastric corpus mucosa, which expresses a type of trefoil peptide, the spasmolytic polypeptide (termed spasmolytic polypeptide-expressing metaplasia or SPEM) and has been shown to be linked more closely to gastric cancer than intestinal metaplasia. Better histological characterization of adenomatous (or type I), hyperplastic (foveolar or type II) and tubule-neck (mucocellular or type III) GED, two-tiered grading system (low and high grade dysplasia) as well as the introduction of Padova and Vienna international classifications of dysplasia seem to be more helpful in GED surveillance and comparative studies. A combination of histopathological features, serum markers such as pepsinogen I, and molecular tests that analyze host susceptibility polymorphisms and bacterial virulence factors, may allow development of strategies for early detection of cancer in the future. At present, pathobiology of gastric cancerogenesis is far from known, despite the progressive knowledge on predisposing environmental conditions and genetic and epigenetic abnormalities, including tumour suppressor genes, oncogenes, microsatellite instability and hypermethylation or the significance of E-cadherin mutational status association with hereditary diffuse gastric cancer syndrome. Recent evidence regarding the importance of several histopathologically derived prognostic factors, such as resection margin status and lymph node metastases and their implications have also been discussed. We aim to review these aspects, with special relevance to gastric cancer specimen reporting. PMID- 21634104 TI - [Operative treatment of primary brain tumors localised in motor zone with direct corticalis electrostimulation--series of 62 patients]. AB - In spite of the progress made in diagnostic procedures and development of the operating rooms technology, considerable neurological deficit after operation of tumors localised in the brain motor zone commits one to direct intraoperative identification of the motor zone. By introducing direct electrocortical stimulation into the routine intraoperative application the primary goal has been achieved -reaching the maximum degree of radicalness of surgical resection while preserving motor centres in the cerebral cortex. METHOD: We are hereby demonstrating a series of 60 patients operated for primary brain tumors localised in the area in the front and around the central sulcus. All operations have been performed under the general anesthetics. During the operations the method of direct electrostimulation (ES) was used for the purpose of identifying motor centres. RESULTS: Intraoperatively a level of subtotal resection was achieved in 22 cases, while radical resection was possible in 38 cases. Significantly higher level of radicalness of surgical resection of the low grade glioma tumor was confirmed statistically in relation to the group of patients with glioblastoma multiforme by applying the ES cortex (p < 0.05). Patients with slow developing brain glioma have statistically considerably higher KI value in relation to the KI values in the group of patients with glioblastoma multiforme (p 0 < 0.01). Difference in the measured values of distance from the coronal suture based on the results of MRI measuring and finding obtained by ES, has shown a statistically considerably higher difference with a glioblastoma multiforme 8.26 +/- 4.288 mm when compared to slowly developing astrocitoma 5.88 +/- 3,080 (p < 0.05). CONCLUSION: Electrostimulation of the brain cortex is a safe, simple and precise method for identification of the brain motor zone which enables prevention of additional postoperative deficit and higher level of surgical radicalness. PMID- 21634105 TI - [Incidence of surgical site infections in a urology department]. AB - OBJECTIVES: To determine the incidence of surgical site infections (SSI) in the urology wards as well as the SSI incidence in relation to the ASA score and surgical site contamination class. MATERIALS AND METHODS: One-year prospective cohort study was conducted in the Urologic department, General hospital, Sabac. The patients were daily examined and their diagnoses were made according to the definition of hospital infections using concurrently the ASA score and surgical site contamination class. RESULTS: Out of 241 operated patients, 32 patients were diagnosed with 33 SSI. The overall incidence rate of patients with SSI was 5.9% while incidence rate of SSI was 6.1%. There were not the differences in the incidence rates according to the ASA score of patients (p > 0.05). The incidence of SSI was 5.0% in the clan wounds, 11.2% in the clean-contaminated, and 20.7% in the contaminated wounds (Chi2 = 8.2 DF = 2 p < 0.016). The patients with SSIs were hospitalized approximately 2 times longer than the patients without SSI (t = -6.28; df = 239; p < 0.001). CONCLUSION: This study suggests that it is necessary to maintain continuous surveillance of surgicical site infections and to implement the preventive measures, especially for clean and clean-contaminated surgery. PMID- 21634106 TI - [T lymphocyte function in laryngectomized patients]. AB - AIM: Malignant tumors of the larynx account for 2.3% of all malignancies, while their frequency among tumors of the head and neck ranges between 12% and 20%. Research on the general immune competence in patients with malignant diseases has provided useful insight in the relationship between immune disorders on one side and the clinical course on the other. Unfortunately, only few complete studies have been published so far with this regard in patients with malignant tumors of the larynx, and therefore our study was essentially aimed at establishing of general immunocompetence, presence and levels of the possible immune disorders and their association with the malignant tumors. MATERIAL AND METHOD: The study included forty two patients with primary squamocellular laryngeal cancer. All the patients underwent surgery, out of whom fifteen were treated postoperatively with radiotherapy. We tested the immune competence prior to the operation and in the postoperative period nine months later. In the venous blood we examined T lymphocyte function, monocyte levels and mononuclear phagocyte function. RESULTS: Preoperative evaluation of the presence and levels of general immune competence in patients with laryngeal cancer, showed a distinct decrease in the proliferative response to the PHA mitogen in vitro, with a tendency to normalize in patients who do not develop a relapse of the disease or distant metastasis during the follow-up period. During the whole study period, the number of monocytes and mononuclear phagocyte activity was above the normal level. CONCLUSION: The patients with operable laryngeal carcinoma had considerable immune disorders at various levels, primarily at the level of T lymphocytes. Of all the disorders, reduced mitotic activity of T lymphocytes in response to mitogens showed the highest dependance on the presence of malignant tissue in the organism. PMID- 21634107 TI - [Length of hospitalization of patients with ankle fracture]. AB - INTRODUCTION: Fractures of ankle are one of the most frequent interacticular fractures that require operative treatment. During the work, the influence of some particular factors (the length of the preoperative period, the complications of the operative period and the application of antibiotics) to the length of the postoperative intrahospital stay, are scrutinised. METHOD: The patients with ankle fracture treated by operation were comprised by the retrospective study in the Traumatologic department in the CHC Zemun in period of 2003 to 2006, and they were divided in three groups depending on the length of postoperative stay. RESULTS: The period of time before the operation (Chi = 0.405, p < 0.01), the appearance of complications (Chi = 0.465, p < 0.01), as well as the length of the period of antibiotic application (Chi = 0.580, p < .01), significantly influence to the length of the postoperative intrahospital stay. The everage length of intrahospital stay for the patients with registered complications was 19 days, while for the patients without registered complications was 10 days. There is statistically significant difference in the length of intrahospital recovery, depending on various complications (logrank = 35.74; df = 5; p < 0.01). CONCLUSION: It is necessary to treat these fractures as soon as possible, for this way of medical treatment results with less number of complications, shorter stay of patients in hospital and thereby reduced treatment costs. PMID- 21634108 TI - [Carpal tunnel in complex regional pain syndrome: a case report]. AB - INTRODUCTION: When treating complex regional pain syndrome (CRPS) a positive outcome can be expected when the condition is identified and therapy is begun in the first six months. Nevertheless, symptoms may not improve by standard treatment as expected. One of the possible reasons for this is the coexistence of associated compressive neuropathies in the affected hand. CASE OUTLINE: A 50-year old female patient with history of CRPS on the left hand which developed as a complication of lateral epicondilytis treated with immobilization is presented. She continued to suffer from intensive pain in her hand despite pharmacological pain relief and initiation of a complex rehabilitation program. Electrophysiologic testing revealed the coexistence of conjoined severe carpal tunnel syndrome. After she underwent surgical decompression pain relief occurred and overall functional results improved. CONCLUSION: Failure of therapy response in CRPS should alert the clinican to check for associated compressive neuropathy. Detection of this complication can prevent further damage of the nerve, hasten recovery of CRPS, and prevent eventual permanent disability of the hand. PMID- 21634109 TI - [Legal aspects of medical errors]. AB - Healing people and medical care are together highly organized technological system with significant expert, ethical and legal regulative. Taking medical care is very sensitive area and it interfears deep into one's integrity, so the law is necessary in this area as a regulator. The aim of work is to show medical errors from legal aspects and clinical practice. Errors, negligent conduct during the medical treatment and bad results of medical treatment are categories that can easily be switched or can sublime themselves into the same thing. That is why correct differentiation of medical errors and viewing every way of medical errors appearance is necessary. PMID- 21634110 TI - [Molecular-genetic markers in lung cancer diagnostics]. AB - The major approaches to different lung cancer marker development are outlined in the review, including genetic, epigenetic, protein, transcryptomic, proteomic, metabolic, and miRNA markers. As far as epigenetic changes are among the earliest events in malignant transformation, methylated markers are thoroughly discussed. Special attention is given to minimally invasive tumor markers, which could be detected in easily accessible biological fluids, because they can be useful for screening and early diagnostics of cancer (before its clinical manifestation) as well as for verification of standard methods of diagnostics. Extracellular nucleic acids, circulating in blood (cirNA), are highlighted as the potential source of material for the early lung cancer diagnostics, prediction of antitumor treatment efficiency, post-treatment monitoring and disease prognosis. PMID- 21634111 TI - [Novel reference gene RPN1 for normalization of quantitative data in lung and kidney cancer]. AB - Quantitative methods of gene expression analysis in tumors require accurate data normalization, which allows comparison of different mRNA/cDNA samples with unknown concentration. For this purpose reference genes with stable expression level (such as GAPDH, ACTB, HPRT1, TBP) are used. The choice of appropriate reference genes is still actual because well-known reference genes are not suitable for certain cancer types frequently and their unreasonable use without additional tests lead to wrong conclusions. We have developed the bioinformatic approach and selected a new potential reference gene RPN1 for lung and kidney tumors. This gene is located at the long arm of chromosome 3. Our method includes mining of the dbEST and Oncomine databases and functional analysis of genes. The RPN1 was selected from 1500 candidate housekeeping genes. Using comparative genomic hybridization with NotI-microarrays we found no methylation, deletions and/or amplifications at the RPN1-containing locus in 56 non-small cell lung and 42 clear cell renal cancer samples. Using RT-qPCR we showed low variability of RPN1 mRNA level comparable to those of reference genes GAPDH and GUSB in lung and kidney cancer. The mRNA levels of two target genes coding hyalouronidases--HYAL1 and HYAL2--were estimated and normalized relative to pair RPN1--GAPDH genes for lung cancer and RPN1--GUSB for kidney cancer. These combinations were shown to be optimal for obtaining accurate and reproducible data. All obtained results allow us to suggest RPN1 as novel reference gene for quantitative data normalization in gene expression studies for lung and kidney cancers. PMID- 21634112 TI - [Allelic polymorphism of six microsatellite DNA loci in populations of Sakha (Yakutia)]. AB - The distribution of allele frequencies of six STR-loci (D3S1358, D16S539, THOI, D8S1179, LPL, HUMvWFII), used in forensic practice, was analyzed in populations of Sakha (Yakutia) (three ethnogeographical groups of Sakha (Yakuts), Evenks, Yukagirs, Dolgans, Russians). Significant differences were revealed between the Russians and all other populations by five markers. Values of total discriminative potential (PD) of studied system for each population were estimated. The level of interpopulation genetic differences (F(ST)) was 0.005. On the basis of allelic polymorphism of STR-loci we estimated phylogenetic relationships between populations under study and populations of different regions--East Europe, South Siberia, Chukotka and Kamchatka. PMID- 21634113 TI - [Cloning of genes, purification and properties investigation of recombinant DNA ligases from the thermophilic archaeon Pyrococcus abyssi and Methanobacterium thermoautotrophicum]. AB - The genes encoding of DNA ligases from the thermophilic archaeon Pyrococcus abyssi (PabDNA ligase) and Methanobacterium thermoautotrophicum (MthDNA ligase) were cloned and expressed in Escherichia coli. The activity of purified enzymes was studied by ligation of two oligonucleotides, one of which had preformed hairpin structure. In the used system the maximal output of reaction products for both DNA ligases was observed near 70 degrees C that is explained by substrate thermostability. At stoichiometric ratio of enzymes and substrate the output of a product reaches of plateau at 70-75% of theoretical ones. Investigated DNA ligases showed different thermostability. The half-time life of PabDNA ligase was about 60 min at 90 degrees C. MthDNA ligase was completely inactivated at this temperature during 10 min. Recombinant DNA ligases from P. abyssi and M. thermoautotrophicum possessed high stability during a storage at 4 degrees C. PMID- 21634114 TI - [Preparing of a ssDNA target in a single-round PCR with low-melt excess primer for hybridization with microarrays]. AB - Method of ssDNA preparing in single-round PCR for microarray application is described. The approach is exemplified on genotyping of DARC gene. It is opposed to two-round PCR that consists of separate symmethric and asymmethric stages. Implementation of reaction in single round is achieved by means of low-melt excess internal primer application. The primer do not anneal during symmethric stage but after decreasing of annealing temperature on asymmethric stage. The results indicate effective oligonucleotide microarray genotyping. The approach reduces time requirements and risk of contamination. PMID- 21634115 TI - [Effect of new potential psychotropic drug, 8-(trifluoromethyl)-1,2,3,4,5 benzopentathiepin-6-amine hydrochloride, on the expression of serotonin-related genes in mouse brain]. AB - Study of molecular mechanisms of psychotropic drug action is the main aim of molecular psychopharmacology. New synthetic analog of variacin 8 (Trifluoromethyl)-1,2,3,4,5-benzopentathiepin-6-amine (TX-2153) was shown to produce anxiolytic and anticonvulsant effects on mice. Here the effect of chronic administration of TX-2153 on expression of some serotonin-related genes in mouse brain was investigated. The drug (10 mg/kg, per os, 16 days) was administered to adult males of ASC (Antidepressant Sensitive Catalepsy) mouse strain characterizing by alterations in behavior and brain serotonin system. The expression of genes encoding 1) the key enzyme of serotonin synthesis, tryptophan hydroxylase 2 (TPH2), 2) main enzyme of serotonin degradation, monoamine oxydase A (MAOA), 3) 5-HT transporter (SERT) and 4) 5-HT(1A) receptor was studied using quantitative RT-PCR. TX-2153 significantly reduced m-RNA level of 5-HT(1A) receptor and MAOA genes in the midbrain without any effect on expression of these genes in the frontal cortex and hippocampus. The drug failed to affect expression of TPH2 and SERT genes in the midbrain. The result indicates involvement of the brain 5-HT system in the molecular mechanism underlying the effect of TX-2153. PMID- 21634116 TI - [ABCA1 gene expression in peripheral blood lymphocytes and macrophages in patients with atherosclerosis]. AB - ABCA1 transporter is known to play important role in the cholesterol transport from peripheral tissues. However its contribution in atherosclerosis development remains not completely understood. Using Real Time PCR, a significant reduction of ABCA1 mRNA level in leukocytes of patients with atherosclerosis was determined when compared with controls. Mean ABCA1 expression levels in leukocytes for the group of patients and for the control group are 0.57 +/- 0.28 and 0.93 +/- 0.14 (p = 0.02). At the same time we detected a significant increase of ABCA1 mRNA level in macrophages of patients when compared with controls. Mean ABCA1 expression levels in macrophages for the group of patients and for the control group are 1.32 +/- 0.10 and 0.90 +/- 0.14 (p = 0.014). In summary, we suggest that expression level of ABCA1 gene may contribute to the development of atherosclerosis. PMID- 21634117 TI - [Loop organization of eukaryotic chromosomes and triple-stranded DNA structures]. AB - To study possible involvement of polypurine and polypyrimidine DNA tracks potentially capable to form triple-stranded structures (H-form of DNA) in compactization of eukaryotic chromosomes a search in silico for "complementary" polypurine and polypyrimidine tracks within 12 eukaryotic gene nucleotide sequences was carried out. Polypurine and polypyrimidine tracks (10-11 b.p.) potentially capable of interacting with each other with the formation of triplex structures ("structurizing" regions) has been shown to be located in chromosomal locus of genes, predominantly in introns and flanking regions. In the case of in vivo realization of such DNA-DNA interactions the chromosomal gene domains can be folded into several small loops. An involvement of DNA triplexes in the chromosomal gene loci compactization may be associated with the gene functioning. The analogous analysis carried out for nucleotide sequences of long (LINE) and short (SINE) repeats dispersed over the genome as well as of satellite DNA has demonstrated a fundamental identity between mechanisms of chromosomal encoding and non-coding regions' compactization. PMID- 21634118 TI - [Arf6, RalA and BIRC5 protein expression in non small cell lung cancer]. AB - Evaluation of tumor markers expression pattern which determines individual progression parameters is one of the major topics in molecular oncopathology research. This work presents research on expression analysis of several Ras-Ral associated signal transduction pathway proteins (Arf6, RalA and BIRC5) in accordance with clinical criteria in non small cell lung cancer patients. Using Western-blot analysis and RT-PCR Arf6, RalA and BIRC5 expression has been analyzed in parallel in 53 non small cell lung cancer samples of different origin. Arf6 protein expression was elevated in 55% non small cell lung cancer tumor samples in comparison with normal tissue. In the group of squamous cell lung cancer Arf6 expression elevation was observed more often. RalA protein expression was decreased in comparison to normal tissue samples in 64% of non small cell lung cancer regardless to morphological structure. Correlation between RalA protein expression decrease and absence of regional metastases was revealed for squamous cell lung cancer. BIRC5 protein expression in tumor samples versus corresponding normal tissue was 1.3 times more often elevated in the squamous cell lung cancer group (in 76% tumor samples). At the same time elevation of BIRC5 expression was fixed only in 63% of adenocarcinoma tumor samples. A statistically significant decrease (p = 0.0158) of RalA protein expression and increase (p = 0.0498) of Arf6 protein expression in comparison with normal tissue was found for T1-2N0M0 and T1-2N1-2M0 groups of squamous cell lung cancer correspondingly. PMID- 21634119 TI - [Methylation profiling of the cell cycle regulating genes in placenta of human embryos with chromosomal mosaicism]. AB - To date the investigation of epigenetic mechanisms of cell cycle regulation, ensuring genomic stability maintaining and accurate transfer of hereditary information to daughter cells, is of a considerable interest. Development of the up-to-date molecular technologies allows determining methylation pattern of the whole genome at a single round of analysis. In this work for the first time the epigenetic status of placental tissues of human embryos with mosaic karyotype was studied using the genome-wide Illumina Infinium HumanMethylation27 BeadChip Array (Illumina, USA). The groups of genes, related to the cell cycle and its regulation and containing differentially methylated CpG-sites in their promoter regions, are determined. The methylation level of oncogenes (ARHGEF1, RGF5), tumor-suppressors (APC2, BRACA2, DCC, GRLF1, RB1, TP73, TSPYL2, VHL) as well as genes, participating in chromosome segregation regulation (CNTROB, GMNN, PROCR, TACC1), was changed most frequently. PMID- 21634120 TI - [Probable involvement of 3'-terminal segment of 18S rRNA in translation initiation of uncapped mRNAs in plants]. AB - A possibility of involvement of 3'-terminal 18S rRNA segment in the cap independent initiation of translation on plant ribosomes was studied. It was shown that 3-terminal segment (nucleotides 1777-1811) of 18S rRNA including the last hairpin 45 is accessible for complementary interactions in 40S ribosomal subunits. Oligonucleotides complementary to this segment of rRNA when added to wheat germ cell-free protein synthesizing system were found to specifically inhibit translation of uncapped reporter mRNA coding for beta-glucuronidase, which bears in the 5'-untranslated region (UTR) a leader sequence of potato virus Y (PVY) genomic RNA possessing fragments complementary to the region 1777-1811. It was shown that a sequence corresponding to nucleotides 291-316 of PVY, which is complementary to a major portion of the 3-terminal 18S rRNA segment 1777-1808, when placed into 5'-UTR, is able to enhance translational efficiency of the reporter mRNAs. The results obtained suggest that complementary interactions between mRNA 5'-UTR and 18S rRNA 3'-terminal segment can take place in the course of cap-independent translation initiation. PMID- 21634121 TI - [Effective and specific control of aml1/eto gene expression in acute myeloid leukemia cells by lentivecior-based RNA-interference]. AB - In presented work, new approach for the control of aml1/eto gene expression in t(8;21)(q22;q22)-positive acute myeloid leukemia cells has been developed. The technique is based on using the RNA-interference and lentiviral transduction methodology. Two new lentiviral vector sets for induction of constitutive anti aml1/eto RNA-interference in acute myeloid leukemia cells have been developed and tested. The first set was based on use of artificial microRNAs (miRNAs) and second one was intended for production of short hairpin RNAs (shRNAs). It was shown that Kasumi-1 and SKNO-1 leukemia cells can be efficiency transduced by each new lentiviral vector. Moreover, the percent of modified leukemia cells that may be easily evaluated in multiplicity of infection (MOI) test achieved more than 90% for Kasumi-1 and SKNO-1 cells at MOI 40 and 20, respectively. Comparative study elucidated that the anti-aml1/eto shRNA-based approach induced a stronger knock-down of aml1/eto gene in Kasumi-1 and SKNO-1 cells than the miRNA-based method did. We hope that the proposed approach may become useful instrument for controlling the aml1/eto gene expression in vitro as well as in vivo investigations of function and biological role of the gene. PMID- 21634122 TI - [Molecular dynamics method for proteins with ionization-conformation coupling and equilibrium titration]. AB - A new realization of the constant-pH molecular dynamics simulation method is proposed. Molecular dynamics simulation is performed for a protein in the most probable ionization microstate of the current conformation taking into account the potential of mean force of protein molecule in the water-proton bath in equilibrium titration conditions (MD-pH-ET). It is shown that: 1) the optimal one is the simulation of the protein in the most probable ionization for a given conformation, taking into account the correction ionization potential of mean force, which results in the librium ensemble of ionization states; 2) new method MD-pH-ET allow one to carry out an optimization of protein structure and the total free energy of a protein in the aqueous solution at constant pH, and to calculate the pH-dependent properties. Method MD-pH-ET possesses the unique features: 1) it uses the most precise and computational-effective realization of calculation of the electrostatic energy of a protein in water solution, the model of continuous dielectric media with Poisson equation and the generalized Born method with "ideal" Born atomic radii; 2) it uses the same model of the potential energy surface in the ionization-conformational phase space, both for calculating the potential energy of the protein and atomic forces and for determining the most probable ionization states; 3) it calculates the total free energy of the protein in the aqueous solution in proton reservoir under the conditions of equilibrium titration. The workability of the new method MD-pH-ET is demonstrated for the molecule of protein BPTI. PMID- 21634123 TI - [The structure of the complexes of DNA with chromosomal protein HMGB1 and histone H1 in the presence of manganese ions. I. Circular dicroism spectroscopy]. AB - The mechanisms of interaction of the non-histone chromosomal protein HMGB1 and linker histone H1 with DNA have been studied using circular dichroism and absorption spectroscopy. Both of the proteins are located in the inter nucleosomal regions of chromatin. It was demonstrated that properties of the DNA protein complexes depend on the protein content and can not be considered as a simple summing up of the effects of individual protein components. Interaction of HMGB1 and H1 proteins is shown to be co-operative rather than competitive. Lysine rich histone H1 facilitates the binding of the HMGB1 with DNA by screening the negatively charged groups of the sugar-phosphate backbone of DNA and dicarboxylic amino-acid residues in the C-terminal domain of the HMGB1 protein. The observed joint action of the and H1 proteins stimulates DNA condensation with formation of the anisotropic DNA-protein complexes with typical psi-type CD spectra. Structural organization of the complexes depends not only on the DNA-protein interactions, but also on the interaction between HMGB1 and H1 protein molecules bound to DNA. Manganese ions significantly modify the character of interactions between the components in the triple DNA-HMGB1-H1 complex. Binding of Mn2+ ions causes the weakening of the DNA-protein interactions and strengthening the protein-protein interactions, which promote DNA condensation and formation of large DNA-protein particles in solution. PMID- 21634124 TI - [The nucleotide sequence features of the mature microRNA seem to be responsible for the affinity to human Ago2 AND Ago3 proteins]. AB - Mature miRNA of 20-24 nt in length are the endogenous sncRNA. They programs RISC to regulate functioning of mRNA with complimentary sites for these miRNAs. In case of Ago3 protein present in human RISC miRNAs direct inhibition of translation, whereas in case of Ago2 is in RISC, than mRNA cleavage in the middle of miRNA/mRNA heteroduplex is also possible. Using ACTIVITY system, that we developed earlier, we analyzed published data on miRNA affinity to human Ago2 and Ago3 proteins. We found increase in miRNA affinity to both Ago2 and Ago3 with the increase of the YRHB tetranucleotide abundance near 3'-end of these miRNAs (r = 0.613, alpha < 0.025). We also found that miRNA tendency to bind Ago2 in favor of Ago3 increases with the RHHK tetranucleotide abundance near miRNA center (r = 0.501, alpha < 0.05). Using these two findings we proposed two formulas to predict miRNA affinity to Ago2 and Ago3 proteins based on the YRHB and RHHK abundances within this arbitrary miRNA. Thereby we made reliable predictions of miRNA affinity to these proteins in RISC for both canonical (alpha < 0.00025) and non-canonical (alpha < 0.05) miRNAs in comparison with independent experimental data. PMID- 21634125 TI - [Identification of protein markers for serum diagnosis of cancer based on microRNA expression profiling]. AB - A new algorithm has been developed for bioinformatics search of putative serum markers of cancer, which includes: 1) identification of microRNAs that are most often and most significantly overexpressed in tumors; 2) selection of mRNA targets regulated by microRNAs; 3) identification of mRNA targets encoding secreted proteins; 4) comparative analysis of mRNA transcription levels in normal and tumor tissues. Application of the algorithm led to discovery of seven putative serum markers of colon cancer: ADAMTS14, ANGPT2, CCL7, DEFA5, MMP11, MMP14, and PLAU. Experiments demonstrated that production of two out of seven proteins (MMP14 and DEFA5) is significantly increased in colon tumors vs. normal samples. PMID- 21634126 TI - [Bimodality of the solubility isoterm of histone H1 complexes with short oligonucleotides]. PMID- 21634127 TI - [The effect of extracellular recombinant human heat shock protein 70 (Hsp70) on protein pattern observed after endotoxin-induced macrophage activation]. AB - The protein pattern of mouse macrophage strain (J774) has been investigated using 2D electrophoresis after combined action of bacterial endotoxins (LPS), heat shock treatment (HS) and administration of recombinant human Hsp70. The investigation demonstrated significant protective effect of HS and recombinant Hsp70 treatment applied before LPS introduction. This effect is apparently realized by means of several signal transduction systems. In the course of the investigation, we have identified eight proteins, which exhibited pronounced changes in their synthesis due to combined treatment. The data accumulated may shed light on molecular mechanisms underlying protective antiseptic action of HS and/or recombinant Hsp70 applied before LPS administration. PMID- 21634128 TI - [Radical operation of infected aortofemoral prosthesis using fresh arterial allograft: our mid-term experience]. AB - INTRODUCTION: The mid-term experience with the use of the fresh arterial allografts in the treatment of aortic or aortofemoral prosthetic infection is presented. MATERIAL AND METHODS: Between 2001-2010 24 patients (23 with the infected graft in aortic or aortofemoral position and one with a mycotic aneurysm of the aortic bifurcation) were operated with the use of the fresh arterial allograft. Male/female ratio was 15/9, average age 65.8 (36-81) years. The gastrointestinal comorbidities dominated this cohort. The total of 70 previous vascular operations (1-9; m. 2.9/patient) were performed with the median of 5.8 years between the first and the last procedure. Seven patients had sepsis (29.2%), aortoeneteric fistula occurred in three. Various technical modifications of the aortobifemoral (13), aortounifemoral (8) bypass, aortic and aortoiliac replacement (3) were performed including the sequential distal reconstructions. The arterial allograft was used within 8-48 hours following harvest (the median cold ischemic time of 20 hours) and all patients were given cyclosporine A perioperatively. RESULTS: In-hospital mortality was 20.8% (5/24), twice caused by postoperative hemorrhage from either the aortic anastomosis or the graft necrosis. The remaining deaths were not related to the allograft itself. Two limbs, preoperatively ischemic, were amputated (8.3%). The median follow-up is 4.6 years (3 m.-8 yrs.). The three-years survival was 68.4% and the known causes of death had no relation to the allograft. The late occlusion of the graft limb occurred twice, stenoses within its course twice and three femoral anastomotic stenoses were disclosed. All were treated either surgically or by PTA/stent and the redo procedures' rate has thus reached 20.5% in the mid-term follow-up interval. One graft has shown a slight diffuse dilatation since requiring but follow-up. CONCLUSIONS: Under the conditions of the ABO compatibility tolerance and ongoing postimplantation immunosuppression the shortly ischemic arterial graft helds its anatomic structure and function and within the hostile setting of the previous infection represents a valuable alternative of the surgical treatment of the vascular prosthetic infection in the aortofemoral position or of the mycotic aneurysm. PMID- 21634129 TI - [Current trends in the surgical management of thoracoabdominal aortic aneurysms]. AB - INTRODUCTION: The authors present a group of patients, who underwent standard procedures on thoracic (TAA) and thoracoabdominal (TAAA) aortic aneurysms from 01 01-2009 to 15-09-2010. MATERIAL AND METHODS: During the above time period, a total of 29 patients were operated. The authors employed as many known organ protection procedures as possible, including reduced heparinization, mild hypothermia (32-34 degrees C), sequential aortic clamping cerebrospinal drainage, left heart bypass (a biopump) with non-ischemic canylation of the femoral artery, selective visceral blood perfusion (superior mesenteric artery and coeliac trunk) and renal arteries perfusion using cold crystallic solution. For technical reasons, monitoring of somatosensory and motor evoked potentials was used only twice. In the majority of TAAA procedures, the thoracoretroperitoneal approach was used. RESULTS: During the studied period, the overall mortality rate was 24%. In 2009, a total of 16 patients were operated, out of which 14 subjects underwent elective procedures. Two subjects underwent urgent procedures for ruptures and both of them died. In 2009, the mortality rate was 21% for planned procedures. In the following year, from January to September 15, a total of 13 patients underwent surgery, out of whom 11 underwent elective and two urgent procedures. The group's overall mortality rate was 15.4%, the mortality rate in elective surgery patients was 9% . One patient undergoing urgent surgery survived and one exited. Postoperative paraplegia was reported in one subject (3,4%), postoperative dialysis due to postoperative renal insufficiency was used in 6.9% and other postoperative complications occurred in 10,3% of the subjects. CONCLUSION: Mortality and morbidity rates in the surgical management of thoracoabdominal aortic aneurysms remain considerably high, although new procedures of organ protection help to reduce it. These highly specialized procedures require a multispecialty approach and a well- coordinated surgical team, specialized in this problematics. PMID- 21634130 TI - [Changes in the true and false visceral segment lumen sizes following endovascular treatment of type B aortic dissections]. AB - INTRODUCTION: The aim of the study was to assess technical success rates of endovascular procedures in acute and chronic type B aortic dissections and changes in the right (PL) and false (FL) lumen diameters in the visceral segment region during short-term and long-term follow up study periods. METHODS: From 2004 to 2009, the authors performed a prospective study, which included a total of 33 patients with acute and subacute (n = 16; 48.5%) or chronic (n = 17; 51.5%) type B dissections of the descending aorta, with dissections spreading as far as the visceral or infrarenal regions. The patients underwent successful implantations of stent grafts (SG) into the descending aorta. The study group included 7 female and 26 male subjects, at the time of the procedure, their mean age was 59 years, (34-70, the median of 56 y.o.a.). The mean follow up time was 39.3 months (8-68, the median of 41 months). During the study period, one of the patients exited due to another internal disorder, three subjects were converted to open replacements for progressing dilatation of the total diameter in the visceral or subrenal region. The true (PL) and false (FL) lumen diameters were measured at four levels: above the origin of truncus coeliacus (L 1), between the origin of truncus coeliacus and the origin of a. mesenterica superior (L 2), between the origin of a. mesenterica superior and the origin of aa. renales (L 3) and just distal to the origin of aa. renales (L 4). The measurements were performed 1 and 6 months after SG implantations and at the end of the study period. RESULTS: The primary entry was successfully sealed in all the study subjects. Significant widening of the true lumen in the region sealed by the stentgraft was recorded in all the subjects, however, the false lumen did not completely disappear in 3 (9%) patients. The false lumen was completely filled with thrombus down to the celiac trunk level (L 1) within one month in 7 (21.2%) patients, within 6 months in 14 (42.4%) subjects and by the end of the study period in 19 (58.6%) patients. The true lumen continued to enlarge at all the measured levels. The most significant enlargement was recorded at L 1 during the first postoperative month (the mean change of 5.9 mm). Furthermore, narrowing of the originally patent false lumen was observed as well, with the most significant change at L1 level during the first postoperative month (the mean change of 6.5 mm). At six months and during the whole follow up study period, further increases in the right lumen diameter and in the total aortic diameter were recorded at all the measured levels. At the same time, the false lumen diameters at all the measured levels continued to narrow, if patent at all. CONCLUSION: False lumen thrombosis along the extent of SG was recorded in 30 (91%) patients, SG filled the lumen completely and the false lumen disappeared. The true lumen expanded at all the measured visceral segment levels. The change was most significant during the first postimplantation month, and the true lumen expansions and the false lumen narrowing proceeded over the whole follow up study period, however, the progression was slower. The visceral segment true and false lumen changes will be monitored further. PMID- 21634131 TI - [Ischemic colitis following AAA procedures]. AB - INTRODUCTION: Bowel ischemia following the AAA operation is a rare complication. However, it is the most common reason of death after elective AAA operation with the incidence of 5-9% and 30-60% following RAAA operation with mortality rate around 60%. METHODS: Retrospective analysis of 207 patients undergoing elective and emergent AAA operation between 1st January 2007 and 31st July 2010 was made. We evaluated clinical and para clinical symptoms leading to the observed diagnosis. RESULTS: During the observed period we have treated 207 patients with the AAA. 174 (84%) patients were elective, 33 (16%) were urgent (RAAA). Postoperative mortality of the entire group was 19 patients (9.2%). The most common reason of death was found to be ischemic colitis--8 patients (42%). Ischemic colitis was diagnosed in 17 patients (8.2%) with the mortality 41.2%. Diagnosis was made by colonoscopy in 58.8% of patients, the rest was operated on following the clinical finding of an acute abdomen. In a group of patients with an early diagnosis and treatment of ischemic colitis (until the 4th post operative day) the mortality rate was 20%, increasing till 71.4% when the diagnosis was done later than on 4th postoperative day. DISCUSSION: Nevertheless the ischemic colitis is a rare complication following AAA operation the surgeon must be proactive regarding this severe complication especially in high risk patients. Clinical symptoms are not specific, most often too early peristaltic and diarrhoea with addition of blood is apparent. Procalcitonine and D-lactate seem to be promising among the blood markers. Colonoscopy is a method of choice if there is any doubt in postoperative process. The need of relaparotomy is given by colonoscopy finding together with clinical, biochemical symptoms, signs and the clinical status of the patient. The resection of diseased part of bowel with the colostomy is mandatory. CONCLUSION: The final diagnosis of ischemic colitis is difficult and the decision for reoperation is a matter of surgeons' experience despite of clinical and paraclinical findings. Only fast and adequate treatment can be successful. Any delay in diagnosis is fatal for the patients. PMID- 21634132 TI - [Plasmatic levels of proinflammatory cytokines in abdominal aortic aneurysms]. AB - INTRODUCTION: Inflammation within the abdominal aortic wall is generally considered a very significant ethiopathogenic factor in the development of abdominal aortic aneurysms. Proinflammatory cytokines are important mediators of inflammation within the abdominal aortic wall. AIM: The aim of the study was to research, whether plasmatic levels of certain proinflammatory cytokines, which can commonly be evaluated (TNFalpha, IL-1, -2, -6 a -8), play a significant role in the development of AAA. METHOD: The prospective non-randomized study included 345 patients with AAAs. The patients were assigned to 5 subgroups based on their symptoms and AAA diameters. The first subgroup included patients with symptomatic AAAs, including AAA ruptures (N = 69), the second subgroup included subjects with asymptomatic AAAs (N = 276) with AAA diameters up to 5 cm (N = 72), the third subgroup included 5 cm (N = 72), the fourth included 5-8 cm (N = 192) and the fifth subgroup included subjects with AAA diameters of more than 8 cm (N = 81). The mean age of patients was 74.1 +/- 7.8 years (56-84 y.o.a.). The male to female ratio was 5:1. The control group included 30 healthy volunteer subjects of similar age and male to female rates, who had no clinical signs of arterial disorders. Plasmatic levels of cytokines were evaluated from venous blood samples using ELISA (Bender, Austria) testing. Statistical assessment of the results was performed using ANOVA and Wilcoxon tests with Spearman's correlation. P values < 0.05 were considered significant. RESULTS: Plasmatic concentrations of proinflammatory cytokines were found to be statistically significantly higher in patients with AAAs compared to those in healthy volunteers. Plasmatic IL8 levels were significantly decreasing proportionally to decreasing AAA diameters (p < 0.05). TNFalpha levels were found to be significantly low in symptomatic patients with AAA ruptures (p < 0.05). CONCLUSION: The study confirmed the significance of proinflammatory cytokines levels monitoring in AAA patients. The authors showed that, for instance IL8 activity and to a certain extent TNFalpha activity, is the highest in small and developing AAAs. These findings would be significant for customized medication therapy aimed at blocking the effects of these factors on the inflammatory process within the AAA wall. PMID- 21634133 TI - [Robot-assisted vascular reconstructions in the aortoiliac region]. AB - OBJECTIVES: The objective of the study was to evaluate the results of the robot assisted laparoscopic aortoiliofemoral reconstructions in a group of 45 patients. MATERIALS AND METHODS: The procedure is performed using three surgical techniques -conventional for the dissection of the vessels in the groin and distal anastomosis creation; laparoscopic for the dissection of the abdominal aorta and robotic for the proximal anastomosis creation. Between May 2006 and May 2010 we operated on 45 patients (37 males, 8 females) at a mean age of 57 years for aortoiliac occlusive disease using robot-assisted laparoscopic approach. We assessed the perioperative and hospitalization data. RESULTS: We created 21 aortobifemoral, 22 aortofemoral and 2 iliofemoral bypasses. The median time of the proximal anastomosis creation was 23 minutes (range 18 to 50 minutes), median clamping time was 60 minutes (range 40 to 95 minutes), median total operating time was 295 minutes (range 180 to 475 minutes). The median estimated blood loss was 265 ml (range 50 to 1200 ml). The median ICU stay was 2 days (range 1 to 8 days). During the follow-up period (median 19 months; range 2 to 50 months), we observed 3 early occlusions (6.7%)--all due to an insufficient outflow. All occlusions were resolved by the extension of reconstructions. No renal, cardiac or pulmonary complications were observed. The 30-day mortality was 0%. Late complications: port-site hernia (2.2%). The secondary patency was 100%. CONCLUSIONS: The da Vinci robotic system allows a precise vascular anastomosis creation and blood loss minimization. The achieved results (clamping time, anastomosis time) are fully comparable to conventional vascular surgery. The robotic system eliminates the technical limitations of laparoscopic aortic anastomosis creation. Robot-assisted laparoscopic aortoiliofemoral bypass grafting seems a safe method with a low complication rate. PMID- 21634134 TI - [Effects of infrainguinal bypasses on long-term patency in pelvic procedures--the significance of hybrid procedures]. AB - OBJECTIVE: The hitherto published data did not provide clear answer to whether the adjustment of poor runoff through infrainguinal by-pass can improve long-term patency of iliac interventions in patients with multiple-stage arterial lesion. Our intent was to respond to this question. MATERIAL AND METHODS: In the period since 1st January 2003 until 31st May 2005 we evaluated two groups of revascularized patients with similar angiographic affection of the iliac and femoropopliteal arteries and statistically comparable input parameters. In the first group incorporating 38 procedures the vascular surgeon performed hybrid intervention (group 1)--one-step iliac intervention with infrainguinal ipsilateral bypass, in the second group incorporating 43 patients angioradiologist performed single percutaneous iliac intervention in catheterization laboratory (group 2). RESULTS: Observation median was 71 months (60-86). Three-year primary patency of iliac interventions was 83% in the hybrid group (group 1) and 72% in single angioplasty group (group 2). Five-year patency was 77% in group 1 and 69% in group 2. In secondary patency the three-year and five-year patency was 94% in group 1 and 83% in group 2. Statistically we did not prove that poor infrainguinal runoff is negative factor of primary patency of iliac intervention (K-M, log rank test, p = 0.58628), as well as secondary patency (p = 0.11474). CONCLUSION: From the long-term perspective it is not possible to consider poor runoff to be independent risk factor of patency of iliac intervention. Infrainguinal bypass within the hybrid procedures shall not improve the results of iliac intervention. PMID- 21634135 TI - [Venous interposition graft in the management of bleeding from distal anastomosis of pedal bypass]. AB - The authors present a rare complication of limb revascularization using pedal bypass, disruption of the surgical wound resulting from infectious etiology with bleeding from the distal anastomosis. The wound disruption was successfully managed by venous interposition grafting. The bypass patency, as well as full functioning of the extremity, were preserved. PMID- 21634136 TI - [Intraarterial peroperative thrombolysis in therapy of the acute limb ischemia]. AB - Acute limb ischemia is serious situation, jeopardizing limb salvage, causing embolization or thrombosis of the native arteries or vascular graft. Immediate surgical revascularization with balloon thromboembolectomy is indicated in patient with acute ischemia stage IIB of the Rutherford classification. Unfortunately, operative thromboembolectomy is frequently incomplete, with persistent thrombus remaining in the majority of patients. Residual thrombus compromises the clinical outcome, leading to poor revascularization and an increased risk of tissue loss. On depending of amount of the residual thrombus after surgical thromboembolectomy, we can choose one of the three options of intraoperative thrombolysis. This article summarized indication and technical realisation of intraoperative thrombolysis and describe cause report of the first patient treated with this method in our country. The combined use of intraoperative thrombolytics following mechanical thromboembolectomy is safe and efficacious method in patients with acute vascular occlusion. PMID- 21634137 TI - [Splenic artery aneurysm as a cause of abdominal pain and hypotension]. AB - Splenic artery aneurysms are the second commonest type of intraabdominal arterial aneurysms. The highest incidence is observed in fertile females. The incidence is higher in multiparae. The mean risk rate of its rupturing is 5% and is higher in pregnant subjects. In case of rupture, the mortality rate is as high as 75% for the mother and up to 95% for the foetus. The authors present a case review of a young female and a middle-aged male subject. In the case review, the authors show a relatively high incidence rate of the disorder and describe the disease course depending on the hospital technical equipment. The differential diagnostic presentation points out disorders which may potentially mimic the splenic artery aneurysm symptoms. The authors discuss the principal role of sonographic and CT examinations in the diagnosis of aneurysms. They also mention the importance of endovascular methods in the treatment of the disorder. At the end of the article, the authors stress out that the incidence of the splenic artery aneurysms is higher than commonly expected and that the diagnosis should be considered in all abnormal cases of left upper and middle abdominal pain. PMID- 21634138 TI - [Cytostatic hyperthermic isolated limb perfusion (HILP) in VFN (General Faculty Hospital in Prague)]. AB - Hyperthermic isolated limb perfusion (HILP) is a standardized method of treatment in selected patients with in-transient locoregional recurrence/methastasis of melanoma or, some other soft tissue tumors (incl. sarcoma etc.) Authors present history and current status of this treatment modality in General University Hospital in Prague. During one year period (7/2009-6/2010) 10 patients were indicated for this procedure. We performed 13 procedures (3x redo), 11 in lower extremity and 2 in upper extremity. There was no serious complication in this cohort of patiens. Multidisciplinar approach is indicated in melanoma patients care. PMID- 21634139 TI - [Simultaneous carotid endarterectomy and coronary revascularization: indications and results]. AB - AIM: Evaluation of operative results and complications in high-risk patients who underwent combined carotid and coronary revascularization. PATIENTS AND METHODS: Combined operation--carotid endarterectomy (CEA) and coronary artery bypass graft (CABG) was performed in the period 2000-2009 in 68 patients. Simultaneous operation was indicated in patients with unstable angina pectoris and 1. symtomatic internal carotid artery (ICA) stenosis > or = 50%, or 2. bilateral asymptomatic ICA stenosis > or = 60% or 3. asymptomatic ICA stenosis > or = 60% combined with contralateral ICA occlusion. Combined operations represented 5.8% of whole CEA series. Mean age was 69.9 (51-82) years, men were 46, women 22. Carotid angiography proved unilateral (always symptomatic) ICA stenosis in 25 patients, bilateral ICA stenosis in 35 patients and ICA stenosis combined with contralateral carotid occlusion in 8 patients. Neurological preoperative symptomatology: TIA was present in u 20 patients, minor stroke in 6 and major stroke in 5 patients. 37 patients were asymptomatic. One CABG was performed in 5 patients, 2 CABG in 20 patients, 3 CABG in 19 patients and 4 CABG in 6 patients. The rest of 18 patients had CABG operation combined with valve procedure. Comorbidity: hypertension 100%, diabetes mellitus 57.3%, hyperlipidemia 60.3%. Shunt was selectively used in 4.4%. The need for shunt was established using back stump pressure and near infrared spectroscopy. RESULTS: Mortality was 8.8% (6/68). The cause of death were multiorgan failure in two cases, ipsilateral stroke in two patients, respiratory insufficiency and cardiac failure due to graft occlusion both in one patient. Good recovery was recorded in 91.2%. CONCLUSION: Combined carotid and coronary revascularization has acceptable neurological morbidity/mortality in high risk patients. Strict requirement is thorough selection of patients. PMID- 21634140 TI - [Simple method of infected vascular prosthesis removal]. AB - INTRODUCTION: Removal of infected vascular prosthesis from aortofemoral or femoropopliteal positions may be technically demanding due to their strong fibrous attachment to surrounging tissues. MATERIAL AND METHODS: The authors describe a simple method of vascular prosthesis inversion and removal from the fibrous tissue using instrumentation for tunelization. DISCUSSION: The authors successfully used the method in a patient with firmly attached and thrombotized vascular prosthesis. CONCLUSION: The procedure reduced duration of the procedure and resulted in reduced traumatization of the surrounding tissue. PMID- 21634141 TI - [Two false myths of the 20th century vascular surgery]. AB - Two myths or dogmas have influenced the therapeutic proceeding of phlebological surgeons in the second half of the 20th century, and both did not stopped to wield their influence up to the present time. The first one is the Cockett's theory of the incompetent calf perforators (blow-out syndrome), the second one is the assertion that femoral vein incompetence invariably causes the most serious stage of chronic venous insufficiency. The incorrectness of the theory of incompetent calf perforators was documented by venous pressure measurements, including direct pressure and electromagnetic flow measurements in the incompetent calf perforators themselves, as well as by plethysmographic results after surgical procedures eliminating the saphenous reflux. The pressure measurements showed that incompetent calf perforators did not cause ambulatory venous hypertension in the superficial veins of the gaiter area; directly the opposite happened: the high hydrostatic pressure measured in the quiet standing position in the incompetent calf perforator decreased profoundly during calf pump activity, as soon as the saphenous reflux was interrupted by digital compression. As to the femoral vein incompetence, no direct evidence has so far been presented which would confirm its hemodynamic significance. On the contrary, plethysmographic findings in patients displaying both saphenous and femoral vein incompetence showed that saphenous reflux was the factor which was responsible for the hemodynamic derangement, whereas femoral vein incompetence was hemodynamically irrelevant. PMID- 21634142 TI - [Neurorehabilitation: for whom? With what objectives?]. PMID- 21634143 TI - [Neurorehabilitation for stroke or brain injury patients]. AB - The neurorehabilitation of brain injury patients usually begins at the hospital, during the acute care, where the aim is essentially at limiting the complications and at beginning the reeducation. The patients benefit then mostly from an inpatient and/or outpatient neurorehabilitation. During all these phases, the rehabilitation declines according to its 3 dimensions, namely: reeducation, rehabilitation and reintegration. In all this process, it is important to remain watchful to the invisible handicaps, such as cognitive impairments, post traumatic brain injury complaints, behavioural or mood changes. These represent a potential obstacle to the social and professional reintegration. PMID- 21634144 TI - [Return to work after brain lesions: cognitive and behavioral factors]. AB - Complete or partial return to work (RTW) succeeds in around 40% of TBI and 25% of stroke patients. Positive factors include: (1) post-traumatic amnesia, hospitalization length, age, race, pretraumatic work and socio-educational status, quality of life, (2) premorbid intellectual level, learning, language and attentional, (3) social cognition and deficit's awareness and (4) absence of behavioural impairment. In stroke, RTW is associated with normal neurological and cognitive/communication abilities. Vocational rehabilitation necessitates: (1) cognitive retraining (speed, visuospatial skills and memory), (2) behavioural approach (compensation, organization, abstraction) and (3) adaptation of work profile. Case management models, with early intervention, continuity of care and coordination, increase twofold partial or complete adapted RTW. PMID- 21634145 TI - [Interest and costs of neurorehabilitation of brain injury patients]. AB - The severity of the initial deficit and the improvement in the first weeks are the strongest indicators for a favorable outcome after stroke. Meta-analyses attempt to evaluate the efficacy of neurorehabilitation, but the results are unconclusive due to the heterogeinity of the groups of patients and therapies. However, there is sufficient data to conclude that repetitive, high intensity, task orientated training is efficacious. New approaches (mental imagery, robotics, virtual therapies...) are also useful but are not better than physiotherapy. It is as important to individualize the approach in a multidisciplinary well organised and communicative setting and to treat early complications. Cerebral plasticity is an individualized process and limited in time, so therapy should be regularly adapted and stopped if the deficit remains stable. PMID- 21634146 TI - [Early neurorehabilitation in an acute university hospital: from dream to reality]. AB - The need for an early neurorehabilitation pathway was identified in an acute university hospital. A team was formed to draw up and implement it. A neuro sensorial, interdisciplinary and coordinated therapy program was developed, focused on tracheostomised patients as soon as they were admitted to the intermediate care in neurology and neurosurgery. The impact of this care plan was evaluated by comparing the results obtained with that pertaining to patients treated previously in the same services. The comparison showed a reduction of 48% of the mean duration of tracheostomy, of 39% in the time to inscription in a neurorehabilitation centre and of 20% in the length of stay in the intermediate care. An early neurorehabilitation care program, with an interdisciplinary and coordinated team, reduces complications and lengths of stay. PMID- 21634147 TI - [Neurorehabilitation: social-professional integration in light of the Federal disability insurance (IV/AI) 5th revision]. AB - The new measures introduced as part of the 5th revision of the Federal invalidity insurance (IV/AI) stress social-professional integration. Thus, when work ceases for a lengthy period, AI offices rapidly organize early intervention measures and professional reinsertion in collaboration with the patient and medical team. These measures are especially well adapted to accompany and support neuro rehabilitation so as to place or maintain patients in the labour market. PMID- 21634148 TI - [Cancer in older patients: which assessment before treatment decisions]. AB - Most of the deaths due to cancer affect the elderly population. The numerous and often underestimated co-morbidities, which weaken elderly cancer patients, interfere with the validity of medical decisions, by raising questions related to the true benefits of the suggested radiotherapy or chemotherapy, in the medium or long term. This leads, sometimes wrongly, either to an absence of oncologic treatment, or to excessive or aggressive therapeutic proposals. The review of the literature leads us to suggest that a global geriatric evaluation should systematically be done to oncologic patients over seventy, with the help of proven tools, in order to offer an appropriate and optimal oncologic treatment. PMID- 21634149 TI - [Keep a cool head!]. PMID- 21634150 TI - [Black on white]. PMID- 21634151 TI - [Some thoughts about "supernumerary (s)"]. PMID- 21634152 TI - [When will be the prohibition of sugar and aspartame?]. PMID- 21634153 TI - [The caregiver]. PMID- 21634154 TI - [Changes of regional environment quality pattern in China since 1986-2008]. AB - For further study of regional differences and the pattern of changes in environmental quality in China since 1986-2008, we perform the principal component analysis, standard deviation, Mann-Kendall and cluster analysis on 18 environmental quality indexes in 28 provinces of China in this paper. Those indexes refer to pollutant emission, pollutants treatment capacities and pollutant emission of per unit land area, etc. The paper indicates that regional environmental quality in China has been increased slightly during this period. It can be divided into four stages: 1986-2000, 2000-2001, 2001-2005 and 2005-2008. The overall patterns of regional environmental quality is the West is higher than the East in general, while the environmental quality of the eastern part have been changed somewhat. For more details, the regional environmental quality in China in 1986 is composed of two parts, the eastern part and the western part, while in 2000 and 2001 the eastern part, the middle part and the western part appears as the overall pattern. For the year of 2005, the regional environmental quality in the western is higher than that of the eastern; meanwhile, the eastern can be divided into the northern part, the middle part and the southern part, and the environmental quality in northern part is better than that of the southern part, southern part is better than that of the middle part. This pattern hardly changed in 2008, except that the area with poor environment quality region had expanded. Pollutant emission of per unit land area played as a main factor; yet both the pollutant emission and the reuse of pollutants impacted the pattern specifically. In addition, the national macro policies, the regional policies, the regional economic and the industrial structure can be primary reason for the change of regional environmental quality pattern in China as well. PMID- 21634155 TI - [Chemical characteristic of water-soluble ions in total suspended particles (TSP) at Lijiang winter time]. AB - 25 total suspended particles (TSP) samples were collected at Lijiang city from Dec 2008 to Jan 2009. Analyzed results indicated that the concentration of CU-, NO3-, SO4(2-), Na+ , NH+, K+, Mg2+ and Ca2+ were 16.00, 14.95, 49.40, 3.12, 44.75, 8.65, 29.50 and 183.02 neq x m(-3) respectively. The average total aerosol loading was 349.39 neq x m(-3), and the average concentration of Ca2+ was 52.38% percent of average total concentration, indicated it was the dominant ion in this sampling. The average concentration of detected cations was 3.35 times of anions, suggested the sum of cations generally exceed the sum of anions in our samples, and the main anion was not detected. The average value of SO4(2-)/NO3- was 3.98, showing that the sustained sources have mainly influences on air. According to the correlation coefficients among ions and factor analysis, Ca2+ and Mg2+ in our samples were from dust, and the sources of NO3-, SO4(2-), NH4+ and K+ were related to various combustion processes by human activities. As a whole, the ions in our samples were mainly combined as CaCO3, (NH4)2SO4, MgCO3, KNO3 and a little CaSO4 and Ca(NO3)2. PMID- 21634156 TI - [Character and sources of conventional anion in different water bodies on the summer in Yulong Snow Mountain area]. AB - Different water samples were collected at Mt. Yulong Region located in lijiang city, Yunnan Province from July 22 to July 28 in 2005. The concentration of anion was measured by Dionex-3000 Ion chromatography and Dionex-600 ion chromatography, different water bodies anion composition and concentration in the study area were analyzed by applying approaches of cluster analysis sea salt iron tracer and typical backward trajectories analyze. The results show that the anion concentration of water of altitude higher than 3 180 m takes on natural state and the influence of human activities and foundation rock is less significant. However, human activities and foundation rock exert direct and decisive influence on the anion concentration of water of altitude lower than 3 180 m, with more significant effects than high altitude. Cluster analysis shows that at high altitude anion concentration and composition of water have strong consistency, the consistency of different water bodies in low altitude area anions is weak, indicating the influence factors is more complex in low altitude than high altitude. Sea salt tracer analysis indicates that the average proportion of nonmarine source of Cl- is 65%; NO3- is predominantly of continental origin; nonmarine sources proportion of SO4(2-) is 93.7%. Typical backward trajectories analyze found that the mostly vapor in the region mainly from the Indian Ocean, part of the vapors from the eastern Pacific, the west wind South Branch occasionally along Tibetan Plateau into the study area. SO4(2-) and NO3(-) which from mainly from Southeast Asia and South Asia input with the monsoon circulation. PMID- 21634157 TI - [Relationship between groundwater quality index of physics and chemistry in riparian zone and water quality in river]. AB - Riparian zone hydrology is dominated by shallow groundwater with complex interactions between groundwater and surface water. There are obvious relations of discharge and recharge between groundwater and surface water. Flood is an important hydrological incident that affects groundwater quality in riparian zone. By observing variations of physical and chemical groundwater indicators in riparian zone at the Kouma section of the Yellow River Wetland, especially those took place in the period of regulation for water and sediment at the Xiaolangdi Reservoir, relationship between the groundwater quality in riparian zone and the flood water quality in the river is studied. Results show that, affected by the river and pond water, the highest point of groundwater temperature is near the pond in spring, and near the river in winter; and regulation for water and sediment at the Xiaolangdi Reservoir also affects groundwater temperature in riparian zone, which reaches its maximum at 100 m far from the river bank. There exists a strong zone of nitrification area at 50 m from the river bank, and in this area, the groundwater pH value is lower by 0.2 to 0.4 unit than that of the other regions, with great annual varieties. The turbidity of groundwater is affected by irrigation, which is more obvious than other indicators of groundwater. The turbidity of groundwater and river water increase rapidly during the early phase of flood retreat, and slope stability of river bank is the initial impact of the soil erosion of river bank. Conductivity, chloride and sulfate data show that the range of 50-200 m in riparian wetland is a very important salt accumulation zone, and the width of salt accumulation zone changes with seasons, and this area is also a very important zone of sulfur reduction. The quality of groundwater at 200 m from the river bank is also significantly affected by floods. Physical and chemical indicators of water change strongly in this area. The result indicates that there is a very close relationship between groundwater and surface water, and it is the typical land and water ecotone between groundwater of riparian zone and the river. Rational protection for this region is critical for the conservation of water quality both in the river and groundwater. PMID- 21634158 TI - [Exploration of relationships between phytoplankton and related environmental factors in the Daning River during sensitive period of algal blooms]. AB - To elucidate relationships between phytoplankton and related environmental factors, phytoplankton and environmental parameters of water quality in the Daning River were conducted using multivariate statistical analysis. Most of the investigated physico chemical parameters [water temperature, total nitrogen, nitrate-N (NO3(-) -N), pH and dissolved oxygen concentration (DO)] in Daning River are significantly different among those sampling sites (ANOVA, p < 0.05). The results indicate that the Daning River has a comparatively high spatial and temporal heterogeneity in the phytoplankton community composition (ANOVA, p < 0.05). The maximum cell density was recorded in Dachang and the minimum density was found at Wuxia-kou sites. Multi-algal species blooms were observed in the same time and place with the maximum density appeared. Correlation analysis was performed to illuminate the relationships between algae density and environmental variables, TN (r = - 0.789, p < 0.05), NO3(-) -N (r = - 0.825, p < 0.05) and NO2( ) -N (r = -0.803, p < 0.05) were negatively correlated with algae density. The results indicate that nitrogen soluble nutrients are key regulatory factors for phytoplankton abundance. The results of redundancy analysis (RDA) indicate that main soluble nutrients (TN, TP, NO3(-)-N and NO2(-)-N), suspended substance (SS) and transparency contributed significantly to phytoplankton community composition. Phytoplankton growth could enhance the pH value. The environmental characterizations and sample sites adjacency contributed significantly to phytoplankton community composition. PMID- 21634159 TI - [Assessment and characterization of heavy metals and nutrients in sediments from Taihu Lake]. AB - The concentrations of Zn, Cu, Mn, Cr, Pb, Al, Fe, iron-bound P (Fe-P) and TP in the sediments from Taihu Lake, a eutrophic shallow lake, China, were determined. The results show that the concentrations of heavy metals are higher than those in the earth crust. Particularly, enrichments of Cr and Pb are more conspicuous than other metals. Moreover, the Pb content in all the sample sites increase markedly from bottom to top, which related to the numerous run-off of Pb-containing wastewater and atmospheric precipitation since 1950. Pollution level in Zhushan Bay of heavy metals and phosphorus is much more serious than other lake regions, and heavy metals and phosphorus contents increase rapidly from about 20 cm of the sediment profile upward, which reveal that the Zhushan Bay is a serious sewage discharging area since 1950. Besides, the significant correlation between Fe-P and TP indicates that Fe-P is an important phosphorus form which can release into the overlying water and be ingested by aquatic organism. That is, Fe-P is responsible for the algae bloom in north and northwest region of Taihu Lake. PMID- 21634160 TI - [Characteristics of organic carbon forms in the sediment of Wuliangsuhai and Daihai Lakes]. AB - The characteristics and differences of organic carbon forms in the sediments of the Wuliangsuhai and the Daihai Lakes with different eutrophication types were discussed in the present study. The results showed that the range of total organic carbon content (TOC) in Wuliangsuhai Lake was 4.50-22.83 g x kg(-1) with the average of 11.80 g x kg(-1). The range of heavy-fraction organic carbon content was 3.38-21.67 g x kg(-1) with the average of 10.76 g x kg(-1). The range of light-fraction organic carbon content was 0.46-1.80 g x kg(-1) with the average of 1.04 g x kg(-1); The range of ROC content was 0.62-3.64 g x kg(-1) with the average of 2.11 g x kg(-1), while the range of total organic carbon content in Daihai lake was 6.84-23.46 g x kg(-1) with the average of 14.94 g x kg(-1). The range of heavy-fraction organic carbon content was 5.27-22.23 g x kg( 1) with the average of 13.89 g x kg(-1). The range of light-fraction organic carbon content was 0.76-1.57 g x kg(-1). The range of ROC content was 1.54-7.08 g x kg(-1) with the average of 3.62 g x kg(-1). The results indicated that the heavy-fraction organic carbon was the major component of the organic carbon and plays an important role in the accumulation of organic carbon in the sediments of two Lakes. The content of light-fraction organic carbon was similar in the sediments of two lakes, whereas, the contents of total organic carbon and heavy fraction organic carbon in the sediment of Wuliangsuhai Lake were less than those in the sediment of Daihai Lake, and the value of LFOC/TOC in the Wuliangsuhai Lake was larger than that in the Daihai Lake. The humin was the dominant component of the sediment humus, followed by fulvic acid in the two lakes. The values of HM/HS in the sediments of Wuliangsuhai lake range from 43.06% to 77.25% with the average of 62.15% and values of HM/HS in the sediments of Dahai lake range from 49.23% to 73.85% with the average of 65.30%. The tightly combined humus was the dominant form in the sediment humus of two lakes, and the followed was loosely combined humus. As a whole, the carbon storage of two lakes were all relatively stable, but the values of PQ, LFOC/TOC, the ratio of loosely to tightly combined humus and HA/FA revealed that, in the sediment of Wuliangsuhai, the humification degree of organic matter was lower than that of Daihai, while the activity of humus was higher than that of Daihai, thus the carbon storage is less stable than that of Daihai. PMID- 21634161 TI - [Investigation of 14 selected antibiotics in sediments of the typical cross sections of Tiaoxi River]. AB - The occurrence and distribution of 14 selected antibiotics in sediments of the typical cross sections of Tiaoxi River were analyzed by UPLC/MS/MS. The 14 antibiotics includes tetracycline (TC), oxytetracycline (OTC), chlortetracycline (CTC), doxycycline (DXC), sulfadiazine (SD), sulfamethoxazole (SMZ), sulfamethazine (SMX), norfloxacin (NFC), ofloxacin (OFC), enythromycin-H2O (ETM H2O), roxithromycin (RTM), chloramphenicol (CPC), thiamphenical (TPC) and florfenicol (FFC). The results show that almost all sediment samples are unavoidably contaminated with antibiotics (detection frequency is more than 70%), and the dominant contaminants are TC, OTC, CTC and DXC, which range from 0.1 to 55.7, 0.7 to 276.6, 6.5 to 131.6 and 6.0 to 15.6 microg x kg(-1), respectively. The concentration of TC, OTC, CTC and DXC observed are as high as 1794.2, 9287.5, 1823.6, 1 149.5 microg x kg(-1) dry weight, in the sediments of the main stream around an outfall of a pig farm respectively. This implied that waste water discharge from animal husbandry was probably one of the main sources to the contamination of these antibiotics in the sediment of Tiaoxi River. Besides the four dominant antibiotics, sulfadiazineshould also be concerned due to its high concentration of 251.6 microg x kg(-1) in the sediment around the outfall of the pig farm. PMID- 21634162 TI - [Distribution characteristics of organochlorine pesticides in bank soil samples and estuarial sediment core of the Minjiang River, Southeast China]. AB - The 19 soil samples and one sediment core were collected from near the banks and in estuary of the Minjiang River. The concentrations of OCPs in the samples were determined by GC-ECD with the internal standard method. The results showed that the concentrations of OCPs, HCHs, DDTs in soils near the banks of Minjiang were in the ranges of 4.31-877.80 ng x g(-1) (with a mean 74.13 ng x g(-1)), 1.32 24.01 ng x g(-1) (7.50 ng x g(-1)), 1.67-876.49 ng x g(-1) (66.64 ng x g(-1)), respectively. Through the study of the vertical distribution characteristics of OCPs in the area, the pollution history of OCPs in the Minjiang estuary was discussed. The concentrations of HCHs and DDTs in the core ranged from 0.34-11.72 ng x g(-1) and 1.12-31.04 ng x g(-1), respectively. The results also showed that the soils and cores in the area have polluted lightly by DDTs, but not been polluted by HCHs. Upper section of the sediment core, o ,p'-DDT was the main composition, which indicated that new DDTs may be still used recently in the Minjiang area. The vertical changes of OCPs reflected basically their production and application history in banks of Minjiang River, Fujian Province. PMID- 21634163 TI - [High-resolution research on the NO3- changes of karst groundwater and its responses to the outside environmental variations]. AB - In order to thoroughly reveal the karst groundwater system in response to the external environment, multi-index high-resolution auto-monitoring instruments were used to research the hydrogeochemistry variations of Qingmuguan subterranean stream. The monitoring indicators were pH value, electrical conductivity (EC), water level, rainfall and NO3-. In the observed 6 times rainfall events, the pH value was mainly controlled by acid rain and EC was impacted by the rain chemistry, physical dilution effect of rainfall and agricultural wastewater. NO3- derived from agricultural activities was less impacted by rain chemistry; and its variations were mainly affected by physical dilution effect of rainfall and agricultural wastewater. Under the influences of the R1 rainfall, the rapid changes of EC and NO3- were contrary in generally, but pH value decreased due to the acid rain. In the R2, R3, R4 and R6 rain events, the water level was positively response to the rainfall and the pH value decreased due to the acid rain; however both of the EC and NO3- decreased due to the dilution effect. With the afflux of agricultural wastewater, both of the EC and NO3- were rapidly changed in synchronization within the shortest of 5 h of and the longest of only 27 h and the pH value showed accelerated decline. During the R5 torrential rain events, water level abruptly increased and all of the pH value, EC and NO3- sharply decreased as a result of dilution effect. The groundwater quality was changed because the afflux of agricultural wastewaters in the entire rainfall periods. According to the National Groundwater Quality Standard, P. R. China (GB/T 14848-9), the groundwater quality of the QSS showed the following order in the entire rainfall events: Grade III --> Grade IV --> Grade V --> Grade 1V --> Grade V --> Grade LV --> Grade V --> Grade IV --> Grade III. As a important drinking water source around local inhabitants, the groundwater quality of Qingmuguan subterranean stream was deteriorated to Grade V in rain-fertilize periods. However, the sampling methods in field did show a simple variation curve. Consequently,the results from the research illustrate the need to redesign hydrochemical sampling methods for karat hydrology system, especially under the impact of human activities. PMID- 21634164 TI - [Abilities of acetaldehyde removal in different water-treating techniques]. AB - The effect of acetaldehyde removal by several kinds of water-treating technologies is experienced in laboratory and the results reveal that traditional water-treating processes have hardly any useful results and the absorption of GAC does no better either for its efficiency depending on the GAC service time greatly and with a very short leakage time. However, the micro-biological degradation process in the BAC filter can consume more than 95% acetaldehyde in 13 minutes when the biomass has been fully acclimatized. The acclimation process usually lasts 4-30 h under field conditions depending on the stabilization and maturity of the original biomass on the carbon, but can be shortened remarkably by means of inhibiting the activity of existing biomass or artificially inoculating acclimatized microbe. PMID- 21634165 TI - [Degradation of highly-chlorinated benzenes by nanoscale iron and palladized iron in aqueous system]. AB - Adsorption and degradation of three typical chlorinated benzenes (CBs) were evaluated using commercial nanoscale Fe (nFe) and synthesized palladized Fe (Pd/nFe) via batch experiments. The results showed that the adsorption capacity exhibited significant difference between various conditions, of which its maximum adsorption amount reached with Tris-buffered solution of pH = 6. 8. 1, 2, 4, 5 TeCB can be dechlorinated by Pd/nFe efficiently within 20 hours. The degradation rates were 75% and 92% in pure water system and Tris-buffered solution, respectively. However, nFe seemed to be inactive to 1, 2, 4, 5-TeCB with final degradation rate of about 25%. Meanwhile, Pd/nFe also only had slight degradation capacities to HCB and PeCB when compared to 1, 2, 4, 5-TeCB. 1, 2, 3, 4-TeCB, 1, 2, 3-TCB and 1, 2-DCB are the only products that were detected during the whole experiments, indicating that chlorine atoms of CBs with less steric effect were preferred to be dechlorinated. The degradation can be fitted by pseudo first order equations. The observed rate constants were ranged from 0. 14 4 h(-1) to 0.115 2 h(-1). PMID- 21634166 TI - [Study on removal of surfactant by activated carbon catalytic ozonation]. AB - The anionic surfactant sodium dodecyl benzene sulfonate (SDBS) is employed as the target pollutant; the SDBS degradation in the processes of ozone (O3), activated carbon (AC) and activated carbon catalytic ozonation were studied. The impact of natural organic matter (NOMs) on the degradation of SDBS by catalytic ozonation was also investigated. Results show that: SDBS removal by using activated carbon catalytic ozonation is higher than that of using single ozonation or AC adsorption respectively; and the most effective time is the early 20 min when the concentrations of ozone and SDBS are relatively high. In the first 5 min (general hydraulic retention time), SDBS removals of the ozonation and AC adsorption were only about 17% and 12.2%, respectively; while the removal of catalytic ozonation reached 54.2% which was much more than the total removal of single ozonation and single adsorption, which showed significant efficiency of the catalytic ozonation. The mechanism of catalytic ozonation involves more radicals could be generated by ozone with the presence of AC; or much higher concentrations of both ozone and reactant would achieve, which enhances the reaction efficiency. Compared with the raw AC, AC experienced relatively long time and high concentration ozone treatment showed little effects on its decomposition efficiency of SDBS during catalytic ozonation. NOMs in the water samples decreased SDBS removals by catalytic ozonation; but the removals kept high under relatively low NOMs concentrations. PMID- 21634167 TI - [Researches on factors affecting the removal of carbamazepine by nanofiltration membranes]. AB - The influence factors on removal of carbamazepine (CBZ) in drinking water by nanofiltration membrane was mainly investigated. The effect of CBZ removal by NF270 and NF90 was firstly compared and found that removal efficiency by NF90 with small pore size showed more effectively than removed by NF270 with large pore size. The next experiment focused on the effect of various factors on removal with respect to CBZ initial concentration, pH, ionic strength and water temperature. The results showed that removal efficiency reduced with decreased pH and increased Ca2+ concentration and water temperature. The effect of initial concentration in the range of 50-500 microg/L on flux and CBZ removal was insignificant. It can be concluded that the retention of non-ionic CBZ by loose NF membrane was strongly dependent on the mechanism of steric (size) exclusion. PMID- 21634168 TI - [Study on the electro-generation of H2O2 using an activated carbon fiber]. AB - The comparative study on the electro-generation of H2O2 using an activated carbon fiber cathode and graphite cathode was investigated. The effect of the operating parameters on the H2O2 generation concentration and current efficiency, such as the initial pH, current density and electrolyte concentration, was also evaluated. The results revealed that the activated carbon fiber cathode was more effective compared to the graphite cathode. The maximum value of H2O2 concentration could be achieved with pH 3.00, current density 8.89 mA/cm2 and electrolyte concentration 0.05 mol/L. However, due to the formation of competitive electrode reactions, the current efficiency of this electrolysis system is lower than other electrolysis system. In addition, a new kinetic model was established to well describe the electro-generation of H2O2. The experimental data were fitted well using the kinetic model. PMID- 21634169 TI - [Removal nitrogen of integrated vertical-flow constructed wetland under aeration condition]. AB - Oxygen is an important limit factor of nitrogen removal in constructed wetlands, so it is the key point for improving nitrogen removal efficiency of constructed wetlands that the optimization of oxygen distribution within wetlands. Therefore, oxygen status, nitrogen removal and purification mechanism of integrated vertical flow constructed wetland (IVCW) under aeration condition in summer and winter have been studied. The results showed that both oxygen levels and aerobic zones were increased in the wetland substrates. The area of oxic zone I (expressing with depth) extended from 22 cm, 17 cm to 53 cm, 44 cm, in summer and winter, respectively. The electric potential (Eh) profiling demonstrated that artificial aeration maintained the pattern of sequential oxic-anoxic-oxic (O-A-O) redox zones within the aerated IVCW in winter, while only two oxic-anoxic (O-A) zones were present inside the non-aerated IVCW in the cold season. The decomposition of organic matter and nitrification were obviously enhanced by artificial aeration since the removal efficiency of COD, TN and NH4(+) -N were increased by 12.2%, 6.9% and 15.1% in winter, respectively. There was no significant accumulation of NO3(-) -N in the effluent with an aeration cycle of 8 h on and 16 h off in this experiment. Moreover, we found that oxic zone I was the main region of pollutants removal in IVCW system, and artificial aeration mainly acted to enhance the purification capacity of this oxic zone in the aerated IVCW. These results suggest that aeration is important for optimization and application of IVCW system. PMID- 21634170 TI - [Segregation effect of purification for nitrogen and phosphate pollution in the subsurface flow constructed wetlands]. AB - Three minitype subsurface-horizontal flow constructed wetlands planted with Calamagrostis angustifolia and Phragmites australis and filled with soil and slag were used to investigate the N, P and pH for upper layer and underlayer wetland system by intermission operation. Results demonstrated that TN removal rates in the superstratum of Calamagrostis angustifolia and Phragmites australis wetlands were 0.771 g x (m2 x d)(-1), 1.481 g x(m2 x d)(-1) with 10 days of the hydraulic retention, which were 1.15 and 1.31 times higher than that of underlayer wetland systems, respectively. Simultaneity, TP removal rates in the superstratum of Calamagrostis angustifolia and Phragmites australis wetlands were 1.655 g x (m2 x d)(-1), 6.838 g x (m2 x d)(-1), respectively, which were 1.13 and 1.28 times higher than that of underlayer wetland systems, respectively. The purification ability of upper layer in the wetland system was higher than that of underlayer. A regular trend of pH changes and upstanding buffer ability of wetland system were found. The pH values in the upper layer of soil-slag wetlands were smaller than that of underlayer which was contrary to the soil wetland. The break-point of pH curve indicates the termination of NH4(+) -N reaction in constructed wetland. PMID- 21634171 TI - [Performance of nitrogen and phosphorus removal from municipal wastewater of different C/N ratios using intelligent controlled systems sequencing batch biofilm reactor ( SBBR)]. AB - The laboratory-scale experiments were conducted to treat synthetic municipal wastewater at different C/N ratios, using a developed intelligent controller sequencing batch biofilm reactor (SBBR), which formed alternative aerobic-anoxic environment. The performance of nitrogen and phosphorus removal was investigated under different conditions (T = 25 degrees C +/- 1 degrees C, V(aeration) = 150 L/h, COD = 300 mg/L, TP = 5 mg/L, TN = 30 mg/L, t(operation) = 15 d; TN = 60 mg/L, t(operation) = 20 d; TN = 90 mg/L, t(operation) = 20 d). The average removal efficiency of COD reaches 85.2%, 91.1% and 97.7%, the average removal efficiency of TP achieves 94.1%, 95.9%, 96.7% ,the average removal efficiency of NH4(+) -N reaches 95.8%, 99.2%, 80.0%, and the average removal efficiency of TN are 90.7%, 81.1%, 58.3%, respectively. With the decrease of C/N ratios, the removal efficiency of N decreases significantly, while the removal efficiency of COD and TP increase slightly. In intelligent controlled SBBR, simultaneous nitrification and denitrification and shortened simultaneous nitrification and denitrification occur, meanwhile, the denitrification of the system strengthens with the increasing of C/N ratios. After the influent finished, intelligent controlled SBBR starts to operate in aerobic-anoxic environment. The efficiency of phosphorus removal is high, moreover, the theory of phosphorus removal is different from the conventional theory of release anaerobic and uptake aerobic phosphorus. PMID- 21634172 TI - [Pollution characteristics of platinum group elements in road dust in central urban area of Beijing]. AB - In order to survey platinum group elements (PGEs) contamination in central urban area of Beijing, dust samples were collected from the second ring road in December 2009. The road samples were digested with aqua regia and separated and purified with cation exchange resin, and the resulting solutions were analyzed by inductively coupled plasma mass spectrometry (ICP-MS). The results showed that the concentrations of Pd, Pt and Rh in road dust ranged from 17.40-458.75 ng x g( 1) (126.66 ng x g(-1)), 10.04-182.89 ng x g(-1) (65.25 ng x g(-1)), 4.00-68.04 ng x g(-1) (22.67 ng x g(-1)) respectively. In comparison with international cities, Pd and Rh concentrations in Beijing road dust were middle level and Pt concentration was lower. Pd concentration was rapidly increased in recent years. The concentrations of PGEs in four locations of the second ring road were arranged in the following order: West approximately East > North > South, which were controlled with the traffic intensity. Size partitioning indicated that the 0.125-0.25 mm fraction had the higher PGEs concentrations than other fractions and the < 0.063 mm fraction had the lower PGEs concentration. These data suggest that autocatalyst PGEs flux estimates into the environment will be significantly underestimated if only a fine grain size fraction (< 0.063 mm) is analyzed. PMID- 21634173 TI - [Mechanism of tritium persistence in porous media like clay minerals]. AB - To investigate the mechanisms of tritium persistence in clay minerals, three types of clay soils (montmorillonite, kaolinite and illite) and tritiated water were used in this study to conduct the tritium sorption tests and the other related tests. Firstly, the ingredients, metal elements and heat properties of clay minerals were studied with some instrumental analysis methods, such as ICP and TG. Secondly, with a specially designed fractionation and condensation experiment, the adsorbed water, the interlayer water and the structural water in the clay minerals separated from the tritium sorption tests were fractionated for investigating the tritium distributions in the different types of adsorptive waters. Thirdly, the location and configuration of tritium adsorbed into the structure of clay minerals were studied with infrared spectrometry (IR) tests. And finally, the forces and mechanisms for driving tritium into the clay minerals were analyzed on the basis of the isotope effect of tritium and the above tests. Following conclusions have been reached: (1) The main reason for tritium persistence in clay minerals is the entrance of tritium into the adsorbed water, the interlayer water and the structural water in clay minerals. The percentage of tritium distributed in these three types of adsorptive water are in the range of 13.65% - 38.71%, 0.32% - 5.96%, 1.28% - 4.37% of the total tritium used in the corresponding test, respectively. The percentages are different for different types of clay minerals. (2) Tritium adsorbed onto clay minerals are existed in the forms of the tritiated hydroxyl radical (OT) and the tritiated water molecule (HTO). Tritium mainly exists in tritiated water molecule for adsorbed water and interlayer water, and in tritiated hydroxyl radical for structural water. (3) The forces and effects driving tritium into the clay minerals may include molecular dispersion, electric charge sorption, isotope exchange and tritium isotope effect. PMID- 21634174 TI - [Sorption behaviors of BDE-28 on natural soils]. AB - The sorption kinetics and isotherms of BDE-28 on three natural soils with different soil organic matter fractions (f(oc)) were investigated. The results indicated that a two (fast and slow)-compartment first-order model was more appropriate for describing the sorption kinetic data, compared to a one compartment first-order model, especially in the initial sorption stage within 25 h. The fast sorption was predominant during the whole sorption process from beginning to the apparent sorption equilibrium; while the contribution of the slow sorption to the total sorption amount gradually increased and then achieved a plateau at 49 h or 55 h. The approaching time to the individual sorption capacity for the fast sorption was much shorter than that for the slow sorption. The contribution of the fast sorption to the increase in the total sorption amount of BDE-28 was prevailing at the beginning of sorption process from 2.5 h to 4.5 h; whereas the fraction of the slow sorption became primary at the subsequent stage of sorption process. The fitting results by the Dubinin Ashtakhov (DA) model were comparable with those by the Freundlich model in the range of apparent equilibrium concentration studied. As for the Freundlich model, the nonlinear exponent (n) values of BDE-28 for the two samples with lower f(oc) (0.72%) or higher f(oc) (7.90%) approached to 1.0 (1.03 +/- 0.05 and 1.00 +/- 0.05, respectively), suggesting the linear sorption characteristics in the studied range of apparent equilibrium concentrations of BDE-28; while the nonlinear behavior of BDE-28 for the left sample with medium f(oc) (4.42%) was indicated by its n value less than 1.0 (0.89 +/- 0.04). PMID- 21634175 TI - [Experiment results of conduction, spectral induced polarization and dielectric characteristics for chrome-contaminated soil]. AB - The resistivity, complex resistivity and complex permittivity of the chrome contaminated soil were studied. Under the different pollution concentration and water content in the soil samples conditions, the relations between the resistivity, complex resistivity and complex permittivity of the chrome contaminated soil and water content and the concentration of pollution were analyzed. When adding chrome pollution with different concentrations and water content, the experimental results show that the resistivity and complex resistivity of all the soil samples decreased with the pollution concentration and water content increased; but the phase of complex resistivity, which reflects the soil's capacitance, decreased below the 20 kHz and increase above the 20 kHz frequency. The real part and imaginary part of complex resostivity increased with the increase of pollution concentration and water content. The concentration of chrome pollutions and water content were the two main factor to determine the soil electrical characteristics. PMID- 21634176 TI - [Effect of competitive interference on the biosorption of lead (II) by Hydrilla verticallata]. AB - Using submerged aquatic plant is a cheap and clean technique to remediate heavy metals in wastewaters. Batch experiments were conducted to assess the biosorption characteristics of Pb (II) ions by fresh tissues of Hydrilla verticallata. The biosorption of Pb(II) was examined for single, binary and ternary solutions at different initial concentrations and different pH values. The experimental results showed that the biosorption capacity increased with increasing pH from 2.0 to 6.0. The biosorption value reached 44.65 mg/g when initial lead concentration was 250 mg/L and pH = 5. Both Cu (II) and Zn(II) ions were found to have an adverse effect on the biosorption of Pb(II) for binary and ternary solutions. In Pb-Cu binary metal solution, when both initial concentrations of lead and copper ions were 250 mg/L, the biosorption capacity for lead ions was decreased to 49.29% of that in single lead ion solution. The biosorption equilibrium data for the Pb-Cu binary metal solution fitted the Langmuir competitive model well (R2 = 0.966). The theoretical q(max) value (58.02 mg/g) was in excellent consistent with that obtained experimentally, and the average relative error between calculated q(e) and experimental q(e) values was only 15.6%. Comparison between biosorption of Pb (II) and Cu (II) by H. verticallata in the binary solution could lead to the conclusion that H. verticallata has no preference of Pb(II) over Cu(II). PMID- 21634177 TI - [Extracellular polymeric substances (EPS) of white-rot fungus and their effects on Pb2+ adsorption by biomass]. AB - The extracellular polymeric substances (EPS) of P. chrysosporium and their effects on Pb2+ biosorption were studied. The product, composition of EPS and the effects on Pb2+ biosorption capacity were investigated in lab via flask experiments. The surface changes of mycelium before and after EPS extraction, before and after Pb2+ adsorption were researched by environment scanning electron microscope with energy-dispersive X-ray analysis (ESEM-EDX). Results showed that at 113 h, the maximum yield of EPS was 125.5 mg/L, which contained 46.6% - 54.3% of sugar and 31.2% - 35.1% of protein. The results of control test after EPS extraction displayed a decrease of biosorption capacity of Pb2+ among 2.12 mg/g (113 h) - 7.73 mg/g (41 h). The results of environment scanning electron microscope (ESEM) showed that the EPS extraction affected the cell wall of white rot fungus and the Pb-contained globular particle after Pb2+ uptake, which was very useful for further study on heavy metal biosorption mechanism. PMID- 21634178 TI - [Effect of heavy metal ions on triphenyltin enzymatic degradation]. AB - The influence of different metal ions and different forms of addition on triphenyltin enzymatic degradation was investigated under conditions using enzyme obtained from Klebsiella pneumoniae. The objective of this study is to illuminate the mechanism of enzymatic degradation of triphenyltin (TPhT). The results demonstrated that the strain was able to tolerate K+, Mg2+, CU2+, Ca2+ and Fe3+ at high concentrations. High concentrations of Zn2+ and Fe2+ had some toxic effects on the strain, thus affecting its growth. The endoenzyme activity was enhanced by metal ions such as K+, Mg2+, Zn2+, Cu2+ and Fe2+ at certain concentrations. In the presence of 30 mg/L of Mg2+, the removal percentage of TPhT was up to 77.22%. Fe3+ restrained the enzyme activity at certain concentrations. Adding K+, Mg2+, Zn2+, Cu2+ into medium can promote the production of enzyme, among which Mg2+ demonstrated up to 85.66% of removal percentage of TPhT, suggesting some metal ions at the appropriate concentration range can be used as enzyme activator for the enzymatic degradation of triphenyltin. Metal ions showed no relevant impact on the cell growth and enzyme production. Certain metal ions can only serve as activators of endoenzyme and exhibited no similar effect towards exoenzyme. PMID- 21634179 TI - [Urban industrial contaminated sites: a new issue in the field of environmental remediation in China]. AB - Contamination of urban industrial lands is a new environmental problem in China during the process of upgrade of industrial structure and adjustment of urban layout. It restricts the safe re-use of urban land resources, and threatens the health of surrounding inhabitants. In the paper, the market potential of contaminated-site remediation was known through analysis of spatial distribution of urban industrial sites in China. Remediation technologies in the Occident which were suitable for urban industrial contaminated sites were discussed and compared to evaluate their superiority and inferiority. And then, some advices of remediation technologies for urban industrial contaminated sites in China were proposed. PMID- 21634180 TI - [Some thoughts of the comparison of risk based soil environmental standards between different countries]. AB - Risk-based soil environmental standard is one of the important aspects in contaminated soil management which have already been widely used in many countries. However, because of diversity in geographical, biological, social cultural, regulatory and scientific aspects among each country, there are great distinctions on both titles and values of these soil environmental standards between different countries. Risk-based soil environmental standards and derivation process were introduced and compared in detail. The variability was analyzed and explained through the comparison of sensitivity risk receptors, land utilizations and pathways of exposure among these countries. We suggest that the risk-based soil environmental standards among the developed countries could be classified as target value, screening value and intervention value, which aim to protect soil for sustainable development in the future, to determine whether there is potential unacceptable risk to specified acceptors, and whether further counter-actions should be conducted, respectively. At last, risk assessment of contaminated soils and establishment of risk-based soil environmental standards in China was proposed. PMID- 21634181 TI - [Triad, a new approach for contaminated site management]. AB - Remediation of a contaminated site is typically a time-consuming and expensive process. The traditional multistage approach of site management is an important contributor of this lengthy and costly process. The emergence of new characterization technologies and past experiences have led to the development of a new site characterization and remediation approach-Triad approach. In this paper, the three key components of the Triad approach, namely, systematic project planning, dynamic work strategy, and real-time measurement, were elucidated. The advantages and applicability of the Triad approach were demonstrated with case studies. Based on the currect practices of site characterization in China, recommendations are made to promote the application of the Triad approach. PMID- 21634182 TI - [Potential role of arbuscular mycorrhiza in bioremediation of uranium contaminated environments]. AB - With the increasing demand for new energy, nuclear industry has been developing very fast, and uranium (U) pollution becomes a serious environmental problem especially in the mining area. The discharge of U products and wastes can contaminate soil and water, and finally threaten human health. On the other side, as an environment-friendly biotechnology, the importance of mycorrhizal technology in remediation of polluted environments has received much attention in recent years. Following a brief introduction of the environmental impacts of U contamination, this review summarized the effects of arbuscular mycorrhiza (AM) on U uptake and accumulation by plants based on recent research progresses, suggested possible application of AM fungi in remediation of U contaminated environment, and finally discussed about the perspectives in relevant research area. PMID- 21634183 TI - [Current researches in microbial remediation of arsenic pollution]. AB - Along with the rapid development of industries, arsenic contamination emerges as one of the world's most urgent environmental problems, especially for the developing countries. Microbial remediation of arsenic polluted environments is a key technique in practice, four aspects, i.e., the special adsorption of arsenic by micro-organisms, the transformation of arsenic speciation and the degradation and volatilization of arsenic compounds by micro-organisms, the effects to arsenic contamination of soil by the interactions between micro-organisms and plant roots, and the molecular biological mechanism of bioremediation for arsenic were reviewed in this paper. In the final section of this paper, the outlook of bioremediation for arsenic and the issues and realms which call for more researches in the future were discussed. PMID- 21634184 TI - [Basic principle and impact factors of soil vapor extraction (SVE) technology for remediation of contaminated soils by volatile and semivolatile organics]. AB - Because soil vapor extraction (SVE) is an effective, economic, and environmentally benign technology to remediate soils contaminated by volatile and semivolatile organics, it has been widely used in the remediation of these soils. The objectives of this paper were to introduce the basic principle of SVE and general steps of constructing SVE engineering, discuss major impact factors on remediation efficiency of SVE technology, and describe three kinds of enhanced SVE technologies. Finally, study and application progress of SVE technologies in China was introduced. PMID- 21634185 TI - [Pb pollution on surfaces in a typical coking plant and health risk assessment]. AB - Pb pollution on the surfaces of buildings and devices at a coking plant was studied. The objectives of this study were to determine the Pb content and its spatial variation on the surfaces, and to assess the potential health risk from this Pb. Ninety-four wipe samples were collected from 56 buildings and devices at the coking plant, and their Pb contents were determined. Surface Pb concentrations ranged from 0-538 micro x g/dm2, and concentrations in 78.7% of the samples exceeded the US Department of Housing and Urban Development (HUD) standard (2.69 microg/dm2). By area, the proportion of samples with concentrations greater than the US HUD standard were 78.9% in the office and living zone, 75.0% in the refining zone, 73.7% in the coking zone and 78.6% in the coke-gas zone. Among the various types of surfaces, the coke pusher had the highest average surface concentration of Pb. The average surface concentrations of Ph on common coking plant surface materials followed the order paints > brick > cement > glass, and the concentrations in 94.4% of the wipe samples taken from paint surfaces exceeded the US HUD standard. PMID- 21634186 TI - [Comparative study on determinations of BTEX in soils from industrial contaminated sites]. AB - In order to ascertain BTEX measurements of soils from industrial contaminated sites, static headspace, purge-and-trap and solid phase microextraction (SPME) combined gas chromatography were selected to determine BTEX in the soils. This method of SPME could not be used to analyze BTEX isomers in soils from highly contaminated sites because the high concentration of organic contaminants eroded the SPME probe head. The recoveries for added standard ranged from 95.2% - 98.2% for static headspace-GC and 99.2% - 101.3% for purge-and-trap-GC. When the soil samples contained low concentrations of BTEX (< 60 mg/kg), the concentration determined by purge-and-trap-GC was 12.6% - 37.6% higher than the value from static headspace-GC. For soil samples containing high concentrations of BTEX, the static headspace-GC result was higher than that from purge-and-trap-GC. The correlation trend lines of individual BTEX isomers to total BTEX were similar for both static headspace-GC and purge-and-trap-GC results. PMID- 21634187 TI - [Chemical oxidants for remediation of BTEX-contaminated soils at coking sites]. AB - An enclosed reactor was used to evaluate Fenton reagent, modified Fenton reagent, potassium permanganate and activated persulfate for removal of benzene, toluene, ethyl benzene, and xylene (BTEX) from soil at contaminated industrial coking sites. The results showed that Fenton reagent and modified Fenton reagent were the optimum oxidants for removing BTEX, and these oxidants decreased the concentration of BTEX in soils by 83% and 73%, respectively. The proportion of BTEX volatilized from the soil was < 4% in both cases, and the rates of BTEX removal from the whole system were 80% and 71%. More than 65% of xylene was removed after treatment with Fenton reagent and modified Fenton reagent. In contrast, benzene, toluene were more resistant to oxidation, and ethyl benzene was the most resistant of these compounds. The concentration of BTEX decreased to some extent when permanganate and activated persulfate were used as oxidants. However, the proportions of volatilized BTEX were 83% and 77% for permanganate and activated persulfate, respectively. This indicates that they could stimulate the desorption and volatilization of BTEX, while they were ineffective for removing BTEX from the environment. PMID- 21634188 TI - [Chemical oxidants for remediation of soils contaminated with polycyclic aromatic hydrocarbons at a coking site]. AB - Different oxidants were evaluated for removal of polycyclic aromatic hydrocarbons (PAHs) from contaminated soils at industrial coking sites with the use of simulating device, and the fate of pollutants during the treatment was investigated. Permanganate showed the highest PAHs removal rate (96.2%) of the oxidants studied. Most of the PAHs were oxidized and only < 1% of PAHs volatilized and transferred to the solvent. The removal of PAHs by activated persulfate reached 92%. Modified Fenton reagent could remove 80% of PAHs, and < 2% of PAHs volatilized and transferred to the solvent after treatment with activated persulfate and modified Fenton reagent. The efficiencies of hydrogen dioxide and Fenton reagent for removal of PAHs were both < 60% ,and up to 6% of PAHs volatilized and transferred to the solvent. In summary, permanganate and activated persulfate are the most effective and environment-friendly oxidants for removing PAHs from contaminated soils. The chemical oxidants could easily remove ANY, ANT, BaP, DahA, BghiP and IcdP, but FLE, CHR and Fla were relatively resistant to chemical oxidation. PMID- 21634189 TI - [Enhanced bioremediation of coking plant soils contaminated with polycyclic aromatic hydrocarbons]. AB - Soil samples contaminated with polycyclic aromatic hydrocarbons (PAHs) were collected from Beijing Coking Plant. The purposes were to isolate PAHs degrading bacteria from the soils, determine their appropriate living condition, enrich them and apply them in the enhanced bioremediation of the contaminated soils. Using each of the 16 USEPA priority PAHs as the sole carbon source, PAHs degrading bacteria were isolated using the method of plate streaking and identified by genetic analysis. In total seven species of PAHs degrading bacteria were obtained. When mixed, these bacteria could degrade the 16 (2-6 cyclic) PAHs studied at appropriate concentrations. In the liquid medium, when the total concentration of the 16 PAHs (sigma PAH16) was 17 microg/mL, single bacteria could grow well and degrade the PAHs. However, when sigma PAH16 was 166 microg/mL, the growth and activity of either single PAHs degrading bacteria or a mixture of the seven PAHs degrading bacteria were inhibited. Aiming at the contaminated soils from Beijing coking plant, five treatments were performed, i.e., control (C), addition of nutrient (N), addition of nutrient and PAHs degrading bacteria (N + B), addition of nutrient and surfactant (N +S), addition of nutrient and PAHs degrading bacteria and surfactant (N + B + S). After five weeks of experiment, compared to the C treatment, the mean removal rate of the 16 PAHs in the N + B treatment was increased 32%, and the mean removal rate of the 16 PAHs in the N + B + S treatment was increased 46% (the mean removal rate of the 10 4-6 cyclic PAHs was increased 52%). The addition of PAHs degrading bacteria and surfactant could significantly enhance the degradation of PAHs in the soils. This study provides evidence for the enhanced bioremediation of PAHs contaminated soil for Beijing coking plant and other coking plants. PMID- 21634190 TI - [Biological toxicity effect of petroleum contaminated soil before and after physicochemical remediation]. AB - Toxicity analysis was studied from using seed germination as an ecological indicator, and the earthworm was considered as a suitable biomonitor animal to determine the ecological hazard of polluted soil. The main results are as follows: These crop seeds have significantly different responses to petroleum pollution. Compared with those plants in clean soil, the germination of most crop seeds planted in contaminated soils is obviously inhabited. Soybean, horse bean and maize are the crop affected most adversely. Fortunately, strong endurance is observed for green soybean under 4 different levels of petroleum pollution, and the seed germination rate are all above 90%. When exposed to pollutants, earthworms could be changed obviously on the level of physiology. That might affect the survival and growth capacity of earthworms, and changed population finally. In high petroleum contaminated soil (concentration of petroleum > 30 000 mg/kg) earthworms can only survive about 5 days. The results suggest that petroleum pollution has great poison to earthworms and can kill earthworms finally. Because pollutants make them dehydrate. Even on the low pollution level, the survival time of earthworm is still very short (3 d or so) in the treated petroleum-contaminated soil. Because after a petroleum ether-treated, the nutrients of soil are disposed with the oil, and the organic matter and other nutrients of the soil have a great impact on the survival of earthworms. PMID- 21634191 TI - [Soil organic pollution characteristics and microbial properties in coal mining areas of Mentougou]. AB - Soil micro-ecosystem including organic pollution characteristics, basic physicochemical parameters, and microbial properties was analyzed which contaminated with organic pollutants in coal mining area. Results showed that the organic pollution level in coal mining area soils distributed from 0.4 to 1.5 mg/g dry soil, which was 1. 5-6 times as much as the background sample. Furthermore, the column chromatography and GC-MS analysis revealed that content of lightly components including saturated and aromatic hydrocarbons exceeded 40%, specifically was alkenes (> C15), hydrocarbon derivatives, and a small amount aromatic hydrocarbons. Totally, the components of organic pollutants extracted in soils were similar to which in coal gangue samples, illustrating the source of soil pollution to a certain extent in coal mining areas. The physicochemical factors such as nutrient level and moisture contents were not conducive to the growth and reproduction of microbe except pH level, which might show inhibition to microbial activities. Microbial density of pollutant soils in coal mining areas was totally low, with specific amount 10(4)-10(5) cell/g dry soil and FDA activity 2.0-2.9 mg/(g x min). Generally, the microbial density and activity were decreased as the enhancing pollution level. However, in-depth analysis was needed urgently because of the complex impact of environmental conditions like pH, moisture, and nutrition. PMID- 21634192 TI - [Accumulation of soil arsenic by Panax notoginseng and its associated health risk]. AB - Panax notoginseng is a valued traditional Chinese medical herb. In this study, the arsenic (As) contamination of soil in P. notoginseng plantation area in Wenshan (Yunnan, China) was investigated; the absorption and accumulation of soil As by the P. notoginseng was revealed; and the associated health risk was evaluated. The results revealed that the soil As concentrations ranged between 6.9-242.0 mg x kg(-1). Arsenic concentrations in 48% of the total soil samples were > 40 mg x kg(-1). The As concentrations in 24% of main root samples, 81% of fibrous root samples, 14% of stem samples, 57% of leaf samples, and 44% of flower/fruit samples were greater than the regulation concentration of 2.0 mg x kg(-1). Arsenic accumulation in the main root increased with the soil As concentration at soil As concentrations < 100 mg x kg(-1), but sharply decreased with the soil As concentration at soil As concentrations > 100 mg x kg(-1). With increasing soil As concentration, the total biomass of P. notoginseng and the main root biomass decreased. Calculating with the As concentration in different parts of Sanqi P. notoginseng plants, percent of the average ingestion rates of As with ADI regulated by FAO/WHO showed fibrous root > leave > flower/fruit > main root > stem. Based on the As concentration in the main root, the daily As intake accounted for a mean fraction of 12.83% (maximum 45.87%) of the acceptable daily intake specified by FAO/WHO,and the ratio increased with the increasing of soil As concentration. Arsenic contamination of soil and P. notoginseng at the plantation area of Wenshan should not be neglected, and effective strategies should be adopt to reduce As accumulation in the plant and human health risk. PMID- 21634193 TI - [Effects of combined pollution of lead and benzo[a] pyrene on seed growth of wheat in soils]. AB - Seed germination, root elongation, shoot elongation and ratio of shoot to root of wheat in soils polluted by lead (Pb) and benzo (a)pyrene (B[a] P) with medium-low concentrations were studied to reveal the ecological effects of combined pollution and screen the indicative markers. Results indicated that seed germination was not sensitive to single or combined pollution of Pb or B[a] P. Root elongation was inhibited by single pollution of Pb or B[a]P to different extents. Extensive interactions between Pb and B[a]P occurred to root elongation of wheat, including synergistic-stimulatory effect and antagonistic-inhibitory effect. The joint action was mainly antagonistic. Single pollution of B [a] P had an inhibitory effect on shoot elongation. Under combined pollution conditions, the shoot elongation of wheat correlated well with Pb contents (p < 0.01). B[a] P or the interactions between pollutants had little effect on shoot elongation of wheat. The joint action on shoot elongation was consistently antagonistic. The response pattern of the ratio of shoot to root was similar to the response pattern of shoot elongation. However, the former had better correlation than the latter, indicating it as a more suitable indicative marker for Pb pollution. If lead acetate was employed instead of lead nitrate, longer root elongation, shorter shoot elongation and no effect on ratio of shoot to root were found. Therefore, the forms of Pb salt had significant influence on seed growth of wheat in soils. PMID- 21634194 TI - [Antioxidative enzymes play key roles in cadmium tolerance of Phytolacca americana]. AB - Phytolacca americana L. has the capacity to take up and accumulate to very high levels heavy metals such as Mn and Cd, and is used for phytoextraction of heavy metal contaminated soils. The role of antioxidative enzyme of Phytolacca americana in response to Cd stress is unknown. The 6-week-old seedlings of Phytolacca americana were exposed to half strength Hoagland solution with 200 micromol/L CdCl2 or 400 micromol/L CdCl2 for 4 days. The content of H2O2 and MDA, and electrolyte leakage increased, while the photosynthetic rate decreased, indicated that the oxidative damage induced by Cd stress in Phytolacca americana was one of the metal toxicity mechanism. The activities of SOD and POD increased rapidly with elevated Cd concentration and exposure time, CAT activity was stable in response to 200 micromol/L CdCl2 stress, and increased only at 3 d later upon 400 micromol/L CdCl2, treatment. Suggested that the enzymatic antioxidation capacity played important role in Cd tolerance of hyperaccumulator plant. PMID- 21634195 TI - [Determination the parameters of bacteria transport in soil]. AB - The tested bacterial in this study is the efficient degrading bacteria Chryseobacterium sp. A-3 to methyl tert-butyl ether (MTBE). The bacterial transport parameters during the process of soil bioremediation were described and determined, which included the properties of soils, adsorption isothermal parameters, adsorption dynamics, effective diffusion coefficient and growth coefficients of bacterial. The results showed, compared with other adsorption isotherms, experimental data are correlated reasonably well by Freundlich isotherm models with correlation coefficient of 99.5%. The exponential coefficient is 1.1, which is close to 1. So linear isotherm model can be used to describe the adsorption isothermal process of bacterial in soil. The adsorption constant of Chryseobacterium sp. A-3 value is 0.98 mL/g. The bacteria adsorption dynamics can be well forecasted by the stagnant film theory, which showed the adsorption process is reversible. The microbial reversible attachment rate coefficient is 0. 004 s(-1), and the microbial reversible detachment rate coefficient is 0. 002 s(-1). Based on the membrane cell and fractal dimension model, the diffusion coefficient 3.66 x 10(-6) cm2/s and effective diffusion coefficient 5.18 x 10(-7) cm2/s can be obtained. Simulation of biodegradation process was carried out. The results showed the maximum bacterial growth rate is 0.01 h(-1), the half saturated constant of MTBE is 134 mg/L as well as the biology rate is 0.33. The proposed of determine methods of bacterial transport parameters can be widely used in laboratory and field studies of microbial migration during soil. These parameters are great significance for the model establishment and study of bacteria transport in soil. PMID- 21634196 TI - [Colloid effects on temporal-spatial variability of iron and manganese in shallow groundwater of garbage contaminated sites]. AB - Simulation tank experiment was conducted to elucidate the temporal-spatial variability of Iron and Manganese in leachate pollution plumes of water-bearing media. Colloid effects on transport and transformation of Fe and Mn in water bearing media were determined emphatically. Moreover, the mechanism of Fe and Mn transport and transformation were discussed by the convection-dispersion, dissolution and transport-deposition of colloid. The results show that the total Fe and Mn in leachate pollution plume was 2.82 times and 7.51 times of infiltration leachate due to the dissolution of water-bearing medium. Along the flow direction, Fe and Mn pollution plumes spread, and the central region of plumes gradually widened by the convection-dispersion and dissolution. In the presence of colloid, the average transport velocity of Fe and Mn plumes central axis from 1.17 cm/d and 1.75 cm/d increased to 1.83 cm/d and 2.5 cm/d respectively, colloid had obvious facilitation to the migration of Fe and Mn. PMID- 21634197 TI - [The use of modern microneurosurgical methods and state of the art technologies in the treatment of brain tumors and neurovascular diseases]. AB - Brain tumors and neurovascular diseases represent the most frequent and serious areas of intracranial neurosurgery. The recent advances in microneurosurgery aim at the complete treatment of the lesions (i.e. radical removal of the tumor, total occlusion of the vascular malformation) while respecting the minimal invasivity for the patient and avoiding risks and morbidity of the surgical procedure. The main tools used in order to accomplish this end are partly meticulous application of recent microneurosurgical principles that enable to treat complicated and deep lesions with minimal retraction and risk of injury to nerves and vascular structures, partly the use of contemporary technologies. Neuronavigation and functional neuronavigation facilitate exact preoperative planning and intraoperative orientation, methods based on fluorescence help to display intravascular blood flow or residual tumor in the operative field and intraoperative magnetic resonance allows exact morphological imaging of the intracranial structures during surgical procedure and increases accuracy of navigation. Electrophysiological monitoring helps to increase the safety of the procedure by continuous tracking of selected brain and nerve functions. Tailored combination and cooperation of aforementioned methods maximizes effect for the uneventful outcome. We present review paper on application of these methods and our experiences in the neurosurgical department Na Homolce hospital and show illustrative cases. PMID- 21634198 TI - [Renal cell carcinoma in the coming era of robotic technology]. AB - Renal cell carcinoma is the most malignant urological tumour and the incidence in the Czech Republic is currently highest in the world. Kidney cancer is associated with seven different genes and is characterized nowadays as metabolic disease. Authors present some current facts about advances in diagnosis and new treatment methods. Different findings required individual approach. Diagnosis improved with more sophisticated ultrasonography, computed tomography, magnetic resonance imaging and positron emission tomography etc. Apart from standard surgery new alternative methods of minimally invasive procedures also assert (laparoscopy, robotic surgery, radiofrequency ablation, kryoablation) and are incorporated in current complex treatment strategies. Systemic therapy with new targeted antiangiogenics drugs is a new standard of treatment for metastatic kidney cancer Complex care with integration of local treatment (surgery, radiotherapy, selective embolization, radiosurgery, cementoplasty, vetrebroplasty etc.) and systemic medication with best supportive care is optimal for a metastatic kidney cancer. More accurate diagnostic methods and complex interdisciplinary approach with integration of systemic therapy with new targeted drugs and new minimally invasive procedures represent new possibilities for patients with kidney cancer. PMID- 21634199 TI - [Deep brain stimulation in movement disorders: a Prague-center experience]. AB - Deep brain stimulation (DBS) in the basal ganglia plays an irreplaceable role in the treatment of Parkinson's disease (PD), essential tremor (ET), and some types of dystonia. Electrodes are typically inserted into the subthalamic nucleus (STN), the internal globus pallidus (GPi) and the ventral intermediate nucleus of the thalamus (VIM). Stimulation of the STN is the main objective in PD, GPi in the treatment of dystonia and PD, and VIM stimulation is effective against tremor. In patients indicated for DBS treatment, stimulation eliminates or reduces rigidity (PD), hypokinesia (PD), tremor (ET, PD) and dyskinesias (dystonia, PD). At the Center for Movement Disorders in Prague, DBS has been performed in 127 patients: in 86 patients for PD, 19 patients for dystonia (11 primary generalized dystonia, 5 primary cervical dystonia, 1 segmental craniocervical dystonia), and in 22 patients for tremor (20 ET, 2 other types of tremor). Intraoperatively we have succeeded in eliminating rigidity in 95% of cases of PD, and tremor in 93% of cases of ET. The positive effect of stimulation on dystonia appeared in 95% with a latency of weeks. Transitory intraoperative morbidity occurred in 8%, intracranial hemorrhage in 1%, infectious complications in 6%, and problems with hardware in 7%. PMID- 21634200 TI - [Antithrombotic therapy after heart valve surgery--current evidence and future trends]. AB - Degenerative valve disease is a growing problem because of the rising amount of degenerative heart valve disease and valve replacement operations among the elderly. More than 300,000 prosthetic heart valve replacements are performed each year worldwide and this rate is projected to increase to as many as 850,000 per year by 2050. There is a higher risk of thomboembolic events following heart valve surgery; and anticoagulation and antiplatelet therapies are necessary. The drug therapy selection depends on the type of surgery performed and the individual risk of the patient in any given situation. This article review assesses the benefit and risks of various therapy schemes as recommended by the Czech, European and American Associations of Cardiologists and Cardiosurgeons. PMID- 21634201 TI - [Early repolarisation syndrome and idiopathic ventricular fibrillation]. AB - Syndrome of early repolarisation is a relatively not common electrocardiographic pattern with typically elevated J wave in most of the cases in lead II, III, aVF and V3-V6. There is increasing evidence that the early repolarisation might be associated with increased risk of sudden cardiac death in otherwise healthy individuals. Early repolarisation ECG pattern in inferolateral leads is associated with sudden death in younger otherwise healthy individuals. Identification of this risky group based on pure ECG criteria is still challenging but it must be considered in individuals with family history of sudden cardiac death or cardiac arrest. PMID- 21634202 TI - [Psychological care as a complement of multidisciplinary medical approach to patients before bariatric surgery operation]. AB - Bariatric surgery has complemented the medical care of patients with obesity at Hospital Na Homolce since 2004. Multidisciplinary cooperation becomes essential. The bariatric team consists not only of a surgeon, a nurse and an anaesthetist, but also an internist, a nutritionist, a psychologist, and possibly a psychiatrist. Our article brings contributions of the psychologist in the bariatric team, and describes the possibilities and importance of his psychological intervention and treatment, both preoperatively and postoperatively. The article offers a review of the methods used in the psychodiagnostic examination prior to surgery, and also mentions the risk factors, such as in cases of deeper pathology in eating behaviour, a personality disorder, low probability of the patient's postoperative compliance and so on, which can decrease the effectiveness of bariatric treatment. In cases where there is psychological contraindication for surgery, the psychologist should offer suitable psychotherapeutic or psychiatric alternative treatment. PMID- 21634203 TI - [Minimally invasive operations in vascular surgery]. AB - Minimally invasive surgery provides an attractive alternative compared with conventional surgical approaches and is popular with patients, particularly because of its favourable cosmetic results. Vascular surgery has taken its inspiration from general surgery and, over the past few years, has also been reducing the invasiveness of its operating methods. In addition to traditional laparoscopic techniques, we most frequently encounter the endovascular treatment of aneurysms of the thoracic and abdominal aorta and, most recently, robot assisted surgery in the area of the abdominal aorta and pelvic arteries. Minimally invasive surgical interventions also have other advantages, including less operative trauma, a reduction in post-operative pain, shorter periods spent in the intensive care unit and overall hospitalization times, an earlier return to normal life and, finally, a reduction in total treatment costs. PMID- 21634204 TI - [Significance of radiosurgery for the treatment of meningiomas]. AB - Treatment of intracranial meningiomas in the light of their benignity (90%) should ensure minimal risk of recurrence, avoid new neurological deficit and preserve quality of life. Stereotactic radiosurgery has proved its long-term effectiveness and safety with a low complication rate. It can be applied as a method of choice for meningiomas in locations that are too risky for microsurgical removal, meningiomas that usually do not exceed 3 cm in diameter and in patients, who are not candidates for microsurgery because of associated disease. It is a part of multimodal treatment for recurrence after operative resection or after partial removal of large meningiomas, where radical surgery represents a high risk of postoperative morbidity and mortality. Indication of radiosurgical treatment of intracranial meningiomas should be considered for each patient individually in order to choose methods or their combination with the lowest risk of complications. PMID- 21634205 TI - [Temporal lobe epilepsy in adults and possibilities of neurosurgical treatment: the role of magnetic resonance]. AB - Temporal lobe epilepsy is the most common type of focal epilepsy diagnosed in adult patients. According to the location of seizure generation it is classified as mesial temporal lobe epilepsy and neocortical lateral lobe epilepsy. Diagnosis of temporal lobe epilepsy can be proved by the combination of the clinical manifestation of partial complex seizures, scalp-video EEG monitoring, results of magnetic resonance imaging (MRI) and imaging of interictal fluoro-deoxy-glucose positron emission tomography. Mesial temporal sclerosis is the most common finding on MRI. Temporal lobe epilepsy is the most surgically amenable diagnosis and results of surgery treatments are clearly superior to the prolonged medical therapy; surgical treatment of the mesial temporal epilepsy with mesial temporal sclerosis has the best clinical results. Except for standard microsurgical approaches such as anterior temporal resection and selective amygdalo hippocampectomy, stereotactic thermocoagulation amygdalo-hippocampectomy is provided in our epilepsy centre. This alternative approach has comparable clinical outcome to the standard surgery approaches in 2 years clinical follow ups. MRI is important not only in diagnostic procedures, but also in neuronavigation of surgery approaches, per operation control of the extent of resections and postoperative follow-ups, especially in failed epilepsy surgery. PMID- 21634206 TI - [Nonconvulsive status epilepticus]. AB - The article summarizes basic facts about classification, clinical presentation, EEG diagnostics and treatment of nonconvulsive status epilepticus. PMID- 21634207 TI - [Significance of radiosurgery in the treatment of brain metastases]. AB - Brain metastases are diagnosed in about 30% adult patients with generalized carcinomas, their accompanying severe neurological symptoms cause worsening quality of patient's life. The development of magnetic resonance imaging and stereotactically guided radiosurgery and radiotherapy enable to apply relatively high doses to well defined intracranial target volumes. Progression free survival after irradiation of brain lesions depends on several prognostic factors. Radiosurgery using gamma knife is an effective treatment with low rate of postradiation complications. PMID- 21634208 TI - [Contemporary pharmacotherapy of epilepsy]. AB - The epilepsies are a frequent group of neurologic disorders. We have witnessed significant advances in their medical treatment recently. About 70 % of newly diagnosed patients are rendered seizure-free. Antiepileptic drug treatment has become more complex. The choice of antiepileptic drug is based on seizure type. However, special patient's needs (drug tolerability, toxicity, ease of use and cost and age, sex and commorbidities) should be considered. The article deals with these special situations. In case of seizure persistence epilepsy surgery could be an option. PMID- 21634209 TI - [Diagnostics of sepsis]. AB - Sepsis is a serious high mortality disease. As regards pathophysiology, it is a systemic inflammatory response to infection. Its timely diagnosis strongly influences mortality. The identification of biomarkers is used for diagnosing, monitoring, and prognosis in septic patients. The methods for their measurement cover a range of disciplines such as clinical biochemistry, haematology, immunology, and clinical microbiology. The most frequently used sepsis markers in routine clinical diagnostics are C-reactive protein and procalcitonin. The detection of selected cytokines, mediators, and surface markers in immunocompetent cells gives us further chance for improvement of this diagnostics. The methods of calculating gene expression represent a technology with a potential for the discovery of new biomarkers which would improve both diagnostics and therapy in the sense of personalized medicine. PMID- 21634210 TI - [Pedal bypass occupies an irreplaceable position in the spectrum of vascular surgery interventions]. AB - BACKGROUND: Pedal bypass grafting has been the method of choice at Na Homolce Hospital's Vascular Surgery Department since 2008. METHODS AND RESULTS: During the period from June 2008 to December 2011, 29 pedal bypass procedures were performed in 27 patients. No perioperative mortality was recorded. Early occlusion of the graft occurred in three patients and two patients experienced delayed occlusion. Loss of the limb as a result of graft occlusion was unavoidable in two cases and one amputation had to be performed despite a patent graft. The primary patency rate for the period under review is 82.75%, and we were able to salvage the limb in 89.65% of cases. Follow-up results for our cohort are comparable with those from other centres dealing with the same problem. CONCLUSIONS: Pedal bypass has excellent early and long-term results and makes a significant contribution to reducing the number of major amputations of the lower limbs. PMID- 21634211 TI - [Six-year experience with cardiac surgery in adults with congenital heart disease]. AB - BACKGROUND: Increasing number of adults with congenital heart disease (ACHD) require reoperation or primary repair. To define risk and outcome of operations of ACHD performed in adult cardiac surgery. METHODS: Between 2005-2010 we operated on 459 patients with ACHD, aged 40.4 +/- 15 years. Operations of ACHD represented 9.5% of 4854 cardiac operations in our department. RESULTS: The 30 days mortality was 1.1%, hospital mortality was 1.52%, long-term mortality among discharged patients was 1.3%. Reoperations after surgery in childhood represented 39%, repeated reoperations 9%, operations of 3 or 4 valves 2.6%. Robotic surgery or minithoracotomy was performed in 9%. Pulmonary hypertension was present in 11.3%, NYHA III-IV in 30%. The risk factors of mortality were: NYHA class III-IV (p < 0.001), cyanosis (p < 0.001), severe systolic dysfunction of left (p = 0.006) or right ventricle (p < 0.001).We did not confirm increased risk in older patients, women, pulmonary hypertension, reoperations and operations of 3-4 valves. CONCLUSIONS: Excellent surgical results among older adults with CHD were achieved by the co-operation of paediatric cardiac surgeon within the experienced centre for adult cardiac surgery. The highest risk was found in patients with NYHA class Ill-IV, cyanosis and ventricular dysfunction. PMID- 21634212 TI - [Measurement of exhaled nitric oxide in the diagnosis of asthma]. AB - Currently exhaled nitric oxide is taken for the best inflammatory marker used in diagnosis of asthma. This short communication deals with recent recommendation, our own experience and comments on the importance of exhaled nitric oxide in clinical practise. PMID- 21634213 TI - [George Wald]. PMID- 21634214 TI - [Epstein-Barr virus positive T/NK-cell lymphoproliferative disease: the pathogenesis and target cells of infection]. PMID- 21634215 TI - [TRP channels as novel cellular sensors]. PMID- 21634216 TI - Home, but not alone: evidence-based maternal, infant, and early childhood home visitation. AB - Home visitation services for young and expectant families have the potential to improve child and parent outcomes in a broad variety of ways, but the effectiveness of home visits may depend on the nature, frequency, and duration of these services. The Patient Protection and Affordable Care Act of 2010 (PPACA) created a new federal funding stream to promote the development and implementation of evidence-based home visiting programs. This issue brief provides an overview of the newly established Maternal, Infant, and Early Childhood Home Visiting program, describes existing approaches to home visitation, and discusses the implications of federal funding for state and local practices. PMID- 21634217 TI - Therapeutic education in patients with chronic heart failure: proposal for a multiprofessional structured programme, by a French Task Force under the auspices of the French Society of Cardiology. PMID- 21634218 TI - New materials complement classic treatment methods. PMID- 21634219 TI - Predicting significant fibrosis in hepatitis C patients in Luxembourg using serological markers. AB - OBJECTIVE: The aim of our study was to assess the diagnostic performance of various serological markers and scores for predicting significant fibrosis retrospectively in a population of patients referring to our hospital for liver biopsy and chronic hepatitis C. MATERIALS AND METHODS: Stored serum obtained from 186 patients were tested for a number of biological markers putatively associated with liver fibrosis. Fibrotest and Forns scores were compared with liver fibrosis pathology scored according to the METAVIR system by multiple logistic regression. RESULTS: The prevalence of significant fibrosis was 44%. Aspartate amino transferase (AST) and gamma-glutamyltransferase (GGT) were most correlated with METAVIR staging, followed by platelet counts and alpha2-macroglobulin. The negative predictive value was 77% and 83% and the positive predictive value was 100% and 84% for the Forns score and the Fibrotest, respectively. In multivariate analysis AST, GGT and alpha2-macroglobulin had independent predictive power. CONCLUSIONS: The accuracy of serological markers in predicting significant fibrosis is limited, because approximately two thirds of patients lie into an indeterminate "grey zone". Serological markers might be useful for patients reluctant to undergo liver biopsy but current predictive scoring systems are too inaccurate to replace biopsies in a routine manner. PMID- 21634220 TI - Chemokine receptor 5 polymorphism in myocardial infarction patients from Luxembourg. AB - BACKGROUND: The CC-chemokine receptor 5 (CCR5) is regulating inflammatory pathways and may thus be implicated in the development and progression of heart failure (HF). A 32 base pair deletion of the ccr5 gene, called CCR5delta32, prevents the expression of CCR5 at the cell surface. We analyzed the association between the CCR5delta32 deletion and the risk and severity of myocardial infarction (MI) in a cohort of patients from Luxembourg. METHODS: Using TaqMan allelic discrimination assay, we genotyped a total of 1080 patients undergoing coronary angiography. This population contained 3 groups of patients: controls with atypical chest pain, abnormal stress testing but normal coronary angiography (n = 154), patients with angina who underwent uncomplicated primary coronary intervention (n = 230), and patients with acute MI (n = 696). In MI patients, left ventricular ejection fraction (LVEF) was determined 1-month after MI with echocardiography. RESULTS: The frequency of the CCR5delta32 deletion was 16.3% in the global population, and was similar between controls, patients with angina and MI patients. The deletion was not associated with variations of plasma levels of creatine phosphokinase and troponin T, nor it was associated with LVEF, New York Heart Association class or 2-year mortality. The frequency of the deletion was comparable between MI patients with LV dysfunction (EF < or = 40%, n = 82) and no LV dysfunction (EF > 40%, n = 402). CONCLUSIONS: The frequency of the CCR5delta32 deletion in Luxembourg is similar to that observed in other European countries and is not associated with the risk of developing MI and LV dysfunction. PMID- 21634221 TI - Predictive relevance of clinical scores and inflammatory parameters in secondary peritonitis. AB - HYPOTHESIS: To measure and evaluate clinical scores and various inflammation parameters for providing a better outcome assessment of patients with secondary peritonitis. DESIGN: Prospective study. SETTING: ICU of a university and a university affiliated hospital. PATIENTS: Fifty-six patients with severe secondary peritonitis were enrolled in this study executed within 4 years. MEASUREMENTS AND MAIN RESULTS: Blood samples were taken preoperatively and 2, 6, 8, 12, 18, 24, 30, 36, 42 and 48 hours post operation, thereafter every 12th hour until day 5 respectively once daily until day 14. Etiology of peritonitis, clinical score systems (APACHE II, MOF and SOFA), and 27 mainly with activity tests or enzyme-immunoassays measurable inflammation parameters were simultaneously analyzed and stratified into lethal outcome (n = 11) or survival (n = 45), respectively. The etiological distribution of peritonitis was identical among both groups. Proportion of intraperitoneal fungi, E. coli, and bacteroids was substantially higher during the primary operation in the group with lethal outcome. With increasing significance initial and follow-up APACHE II, MOF and SOFA scores provided higher values in this group. Various plasma/serum parameters of hemostasis, leukocyte proteolytic system, acute phase reaction, cytokine system, cell adhesion, opsonization, and main organ functions showed significantly different values between both groups at the preoperative stage and/or during observation period I (day 0-4). Logistic regression analysis revealed the SOFA score and neopterin concentration as the combination with the best sensitivity (63.6%) and specificity (93.2%) for predicting the patients' survival even at the preoperative stage. For the observation period I, the combination of SOFA score and TNF receptor II showed the highest predictive sensitivity (72.7%) and specificity (95.6%). CONCLUSION: Evaluation of the severity of secondary peritonitis using a scoring system with high prognostic relevance could conceivably result in an earlier and adequate application of intensive care such as hemofiltration, administration of immunoglobulins and serial abdominal lavage to improve successful outcome. PMID- 21634222 TI - Thoroughly modern: hospital addition blends stylish design and quality care. PMID- 21634223 TI - Auditory assistance. Strategies to reduce hospital noise problems. PMID- 21634224 TI - Island of stability. Power system overcomes unique vulnerabilities. PMID- 21634225 TI - 'FGI Guidelines' rolls out. A look at the 2010 edition set for delivery this month. PMID- 21634227 TI - Read and reflect. PMID- 21634226 TI - Working 'outside-in'. Efficient and effective hard floor maintenance. PMID- 21634228 TI - New records of helminth species and their hosts in Poland. AB - Information on species of parasitic helminths native to Poland is provided by the Pasozytnicze helminty Polski. Gatunki. Zywiciele. Biale Plamy. Until the middle of 2007, there were 1205 species of recorded helminths including 126 species of Monogenea, 338 of Trematoda (Digenea), 279 of Cestoda, 427 of Nematoda and 35 of Acanthocephala. They represented 32.9% of the species recorded in Europe (i.e., registered in the Fauna Europaea database). During the following three years up to the middle of 2010, 64 new species of Polish helminths were detected: 7 Monogenea, 15 Trematoda (Digenea), 21 Cestoda, 20 Nematoda and one Acanthocephala. Most hosts of the new helminth species were reported from birds (40 species), but also from fish (13 species) and mammals (10 species). Only one new species of helminth was detected in amphibians. PMID- 21634229 TI - A checklist of leech species from Poland. AB - In this study 47 leech species from Poland are listed. They belong to two orders, two suborders, five families and 17 genera. The checklist also includes the information about hosts, distribution in Poland and references concerning the leech species discussed in this study. PMID- 21634230 TI - Identification of the piroplasms isolated from horses with clinical piroplasmosis in Poland. AB - The study was aimed at determining the cause of the diseases in three horses exhibiting symptoms of fever, ataxia, mucus membrane paleness, haematuria and thrombocytopenia. The PCR technique revealed the presence in the blood of 18S RNA Babesia/Theileria spp. genetic material. DNA amplification using primers RLB F2 and RLB R2 produced 430 bp size products. The sequences of these PCR products demonstrated a 95.6-97.5% similarity with the sequence of the fragment of 18S RNA Babesia equi, gene number DQ287951 in the GenBank. The treatment utilizing the subcutaneous application of the imidocarb resulted in gradual recovery of the diseased animals. PMID- 21634231 TI - The human aural myiasis caused by Lucilia sericata. AB - Myiasis is a rare, worldwide, human disease with seasonal variation, caused by developing larvae of a variety of fly species. It can be dangerous when infestations penetrate into the brain. In the available literature, we have found only a few papers concerning ear myiasis caused by Lucilia sericata. Here, we report 2 cases of aural myiasis. Early intervention (surgical removal, occlusion) in these cases should prevent complications. Larvae, for further examination, should be killed by immersion in very hot water, then preserved in an ethanol. PMID- 21634232 TI - Morphological features of Cloacotaenia megalops (Nitzsch in Creplin, 1829) (Cestoda, Hymenolepididae) from different hosts. AB - The tapeworm species Cloacotaenia megalops (Nitzsch in Creplin, 1829) is characterized by a very specific morphology. A particularly distinct feature is a large almost square scolex in which four fleshy suction cups are embedded and a rudimentary rostellum. During standard studies of wild duck cestode fauna in northwestern Poland, some morphological differences were observed among specimens, mainly in the shape of scolices and hermaphroditic proglottids of C. megalops isolated from different bird species. This paper attempts to demonstrate the impact of the host species on morphology of the parasite. The study material consisted of 39 individuals of C. megalops (19 from Aythya fuligula, 10 from Anas platyrhynchos, 4 from Bucephala clangula, 4 from Aythya marila and 2 from A. ferina). The isolated parasites were fixed and stored in 70% ethanol and solid preparations were made. Selected external and internal structures of the parasite were measured and photographed using a Zeiss microscope, a microscope camera Opta Tech 2.1 and Opta View 6.0.2.2. It was stated that the sizes of the most important elements of the parasite morphology are generally consistent with those presented in the available literature, nevertheless some differences were observed in both morphology and sizes of some anatomical structures, especially in the form of scolices and hermaphroditic proglottids between specimens isolated from hosts representing three different eco-tribes (Aythyini, Anatini and Mergini). PMID- 21634233 TI - Actitis hypoleucos (L., 1758)--new Polish host of the trematodes: Plagiorchis nanus (Rudolphi, 1802) and Leucochloridium perturbatum Pojmanska, 1969. AB - In July-August of 2005-2008, 47 digenean trematodes were examined from 7 specimens of the common sandpiper, Actitis hypoleucos (L., 1758). The digeneans were collected during a study on the food composition of the common sandpiper. The research was conducted near Lisewo Malborskie (near Tczew, Zulawy Wislane, Gdansk sea-coast), at a resting site on the birds' migration route. The foods ingested were extracted from live birds by draining their alimentary tracts with a nasogastric tube. Two digenean species were identified: Plagiorchis nanus Rudolphi, 1802 (Plagiorchiidae) and Leucochloridium perturbatum Pojmanska, 1969 (Leucochloridiidae). Actitis hypoleucos is the host for these new records in Poland. The P. nanus individuals isolated from A. hypoleucos differed in terms of the visible uterus and the extent of the vitellaria in the posterior part of the body. PMID- 21634234 TI - Analysis of clinical symptoms and selected hematological indices in hospitalized children with Ascaris lumbricoides infection from the northeastern region of Poland. AB - Ascariasis is the most common soil-transmitted helminth infection in the world. The objective of this study was to analyze the clinical symptoms and selected hematological indices of ascariasis in hospitalized children from the northeastern region of Poland. Patients in the Pediatric Ward hospitalized in the Regional Hospital in Dabrowa Bialostocka in the period of 2005-2007 were included in this retrospective study. The intestinal stage of ascariasis was diagnosed on the basis of positive coprological survey performed using the decantation technique. A total of 938 patients were included in the study, 1801 stool samples were evaluated, and A. lumbricoides-positive tests were obtained from 252 children. Ascaris-positive young children (< or = 3 yrs) accounted for 3.0% of all hospitalized children, Ascaris-positive preschool-aged children (4-7 yrs) accounted for 8.1% and school-aged children (8-18 yrs) for 15.8%. Seasonal patterns were observed in the prevalence of A. lumbricoides (maximum in August December). There was no relationship between BMI z-score, hemoglobin levels and prevalence of infection with Ascaris lumbricoides. Significant predictors of intestinal stage ascariasis in a multivariate logistic regression model were: abdominal pain as a reason for hospital admission (OR-2.19; 95% CI 1.62-2.95; p < 0.001) and age from 4 to 7 years (OR-2.0; 95% CI 1.41-2.80; p < 0.001). The prevalence rate of ascariasis was not higher in the group of patients with atopic diseases (bronchial asthma, allergic rhinitis, atopic dermatitis) and co-existing ascariasis did not affect the eosinophil counts in the peripheral blood. Ascariasis is still a current pediatric clinical problem characterized by non specific clinical manifestations, which should be taken into consideration in the differential diagnosis of children's diseases. PMID- 21634235 TI - Pierre looks after a patient in post-operative care. PMID- 21634236 TI - Caryospora biarmicusis sp.n. (Apicomplexa: Eimeriidae) infecting falcons from the genus Falco in Saudi Arabia. AB - The oocysts of Caryospora biarmicusis sp.n. is described from the feces of the lanner falcon, Falco biarmicus, from the falcon market in Riyadh City, Saudi Arabia. Sporulated oocysts are ovoid in shape, measuring 40.2 x 34.7 (37.5-42.4 x 32.9-35.7) microm; shape index (L/W) is 1.16 (1.08-1.31) microm. The oocyst wall is smooth and bi-layered. Micropyle and polar granule are absent, but an oocyst residuum is present. Sporocysts are spheroid, 20.1 (18.6-21.3) microm; with a smooth single-layered wall, lacking Stieda body. Sporocyst residuum is present as numerous small granules. Sporozoites are stout with a large single refractile body. PMID- 21634237 TI - A light microscopic study on the haemogregarine species infecting the lizard Acanthodactylus schmidti from Saudi Arabia. AB - Eleven out of 42 (26.2%) lizards Acanthodactylus schmidti were found harboring haemogregarines in their peripheral blood. The erythrocytic stages were differentiated into 2 forms: the young form (trophozoite) measured 12.5 +/- 0.5 x 1.7 +/- 0.3 microm and the large mature form (gametocyte) measured 19.8 +/- 1.7 x 1.7 +/- 0.3 microm. The infected erythrocytes were distorted from 16.0 +/- 1.2 x 9.2 +/- 0.2 microm to 20.2 +/- 1.8 x 6.7 +/- 0.8 microm, hypertrophied and faintly stained. None of the leucocytes seemed to be parasitized by the present parasite. Schizogony took place in the endothelial cells of lung capillaries and parenchyma of liver. Two types of schizonts were recorded; microschizonts of 9.35 x 9.65 +/- 0.38 microm and macroschizonts of 25.36 +/-1.81 x 20.35 +/- 0.82 microm. The microschizonts produced 6-12 merozoites, while the macroschizonts produced 18-36 merozoites, however there were no differences between both merozoites, each measured 11.54 +/- 0.81 x 1.41 +/- 0.71 microm. The infected host cells were markedly hypertrophied with noticeable irregularity and faint stainability as well as some vacuolation, necrosis or shrinkage and necrosis. PMID- 21634238 TI - T-cell immunity in human chronic toxoplasmosis. AB - Toxoplasmosis is a disease affecting 500 million people worldwide. The seroprevalence varies (from 5% to 90%), depending on geographical location, age, habit of eating raw meat or unwashed fruit and vegetables, and general level of hygiene. The incidence of infections is higher in warmer and humid climate and increases with age. The disease can be congenital or acquired. Cell response (TNF alpha and IFN-gamma secreted by Th1 cells) and humoral response (IL-4, and IL-10 secreted by Th2 cells) were evaluated. This study assessed the effect of T. gondii on chosen indices of the immune response. The study involved 45 women infected with T. gondii (aged 18-42 years) proven have a chronic toxoplasmosis (IgG positive & IgM negative). The control group consisted of 25 healthy women (aged 18-45 years) (IgG and IgM negative). The results showed that patients infected with T. gondii had increased production of theTh-1 cytokines involved TNF-alpha and IFN-gamma that responsible for the cellular response compared to controls. Also, increased production of Th2 cytokines involved IL-4 and IL-10 that responsible for humoral response compared to controls. PMID- 21634239 TI - Morphological forms of Trypanosoma evansi from blood of Arabian camel (Camelus dromedarius) in the Riyadh metropolitan areas. AB - Trypanosoma evansi is commonly referred to as haemoflagellate infesting camels and other vertebrates. It is causative agent of number of diseases. T. evansi has several morphological forms. We have detected three forms in the blood of camels, slender, transitional and intermediate. The present study is the first investigation on the morphological forms of T. evansi in Arabian camels in Saudi Arabia. PMID- 21634240 TI - Zoonotic brucellosis: an underestimated or misdiagnosed disease in Egypt. AB - Over six months, 329 suggestive consecutive brucellosis human cases were diagnosed in attending the out-patients clinics of Al-Azhar and Ain Shams Universities Hospitals and Giza Governorate Farmers. They were 100 females and 229 males with ages ranged between 15-65 years old. A total of 213 (64.75%) were working in dairy farm and/or consumed raw milk, 16 (14.85%) used home slaughtering of sheep, and 100 (30.4%) were working in Giza Government slaughter house. Clinically and by ELISA-IgM 259 out of 329 the subjects were proven brucellosis patients (77.8%). Besides, other patients had toxoplasmosis, or schistosomiasis mansoni or fascioliasis. Double infection was encountered with toxoplasmosis and either schistosomiasis or fascioliasis. The causes of endemic liver parasitosis that may give false-clinical diagnosis were excluded. Signs and symptoms of brucellosis patients were fever (91.5%), chills (84.1%), Myalgia (69.5%), headache (58.2%), fatigue (77.2%), anorexia (54.1%), tachycardia (38.6%), hepato-and/or splenomegaly (46.2%), lymphadenopaqthy (19.6%) lower back abdominal pain (8.8%) and/or constitutive symptoms (13.1%). PMID- 21634241 TI - Allergenic Dermatophagoides mites causing asthma among schoolchildren at Ain Shams District, Cairo, Egypt. AB - The present study was performed on house dust samples collected from ten homes of schoolchildren suffering from asthma at Ain-Shams district, over a period of two years (2008 & 2009). The data revealed that the total annual density of the two allergenic mites (Dermatophagoides pteronyssinus (Trouessart) & Dermatophagoides farinae (Hughes) was 202 individual mites with 22 asthmatic children in the first year, and individual mites increased to 268 with 36 asthmatic children in the second year. The two house dust mites were more abundant in bedrooms than in living ones (292 & 187 mites, respectively). The prevalence of the house dust mites on the mattresses and furniture were higher in bedrooms and living rooms than on the floors (340 & 140 mite, respectively). Winter season recorded the highest prevalence for both mites in the first and second year (87 & 110, respectively). Summer represented the lowest values (19 & 25, respectively). PMID- 21634242 TI - Mange mites causing scabies in Egyptian buffaloes at Giza Governorate, Egypt. AB - In the present study, 560 buffaloes from a private animal farm in Giza Governorate were examined over one year for scabies infestation. The results showed that 466 Buffaloes (83.2%) were infested with two mange mites; Psoroptes natalensis and Sarcoptes scabiei. The highest prevalence rate of infestation was during winter (90.4%), and lowest was in summer (71.4%). The infestation on females was 90.4%, while on males was 80.8%. The mites were predominant on the withers, lumber, back, croup and the external angle of ilium. Buffaloes more than five years old were highly infested than smaller ones, but no mite's infestation were detected in those less than one year. PMID- 21634243 TI - Trypanosoma evansi in dromedary camel: with a case report of zoonosis in greater Cairo, Egypt. AB - Trypanosoma evansi (Steel, 1885) Balbiani, is the cause of trypanosomiasis (Surra) which multiples in the blood and body fluids. The incubation period varies from 7-15 days. The mortality rate was up to 20% and fatality rate may reach up to 100% in untreated camels. Three hundred randomly selected camels were 200 males 4-6 years old and 100 females 10-15 years. They were examined clinically and diagnosed by Giemsa stained blood smear, anti- trypanosomiasis antibodies by ELISA and urine Thymol turbidity test for natural infection with T. evani (Surra). The results showed that camels were naturally infected with T. evansi as indicated by stained blood film examination and/or ELISA. Infection in males was 6.0% (stained blood smears), 8.0% (ELISA) and 5.0% (urine thymol turbidity test). In females the infection rate was 9.0%, 24.0% and 12% respectively. By correlation with suggestive clinical manifestations, ELISA proved to be more sensitive and specific (13.3%) than stained blood films (10.0%) and urine Thymol turbidity test (7.3%). Regarding humans, one out of 30 was positive as indicated by ELISA and stained blood smear but was negative by urine thymol turbidity test. The human case was successfully treated as indicated clinically, parasitologically and serologically. This is the first reported Egyptian human case of trypanosomiasis evansi, a neglected zoonosis. PMID- 21634244 TI - Effect of Ficus sycomorus and Azadirachta indica extracts on liver state of mice infected with Schistosoma mansoni. AB - The potential role of hepatoprotective and antipathological effect of Ficus sycomorus and Azadirachta indica extracts was evaluated for scavenging the reactive oxygen species (ROS) and reduced the oxidative damage and pathological changes in the liver of S. mansoni infected mice. The levels of alanine aminotransferase (ALT), asparate aminotransferase (AST) and gamma glutamyl transferase (GGT) were evaluated in the infected mice and treated orally with each plant extract 12 weeks post infection (P.I.) in a dose of 500 mg/kg of each plant extract for five consecutive days and sacrificed two weeks P.I. The infection of mice showed an elevation of ALT, AST & GGT. Treatment of mice with 70% methanol extract of each plant extract reduced significantly ALT, AST & GGT elevation. The highest reduction was with the methanolic extract of F. sycomorus (42%, 35% &44% for ALT, AST & GGT respectively). Fractionation of the methanolic extract of each plant was carried out. The effect of ethyl acetate and butanolic fractions of each plant was also evaluated. The result showed that the two fractions lowered the levels of the tested enzymes and decreased the number and size of granuloma diameters with an increased in the percentage of degenerated ova. PMID- 21634245 TI - Comparative study between PCR and microscopic examination in diagnosing Entamoeba histolytica. AB - Differentiation between E. histolytica and non pathogenic species as Entamoeba dispar, which are morphologically identical species, is essential for rapid treatment decision, precaution of the invasive disease and public health. This study evaluated the real time PCR for detection of intestinal amoebiasis in comparison with microscopic examination. Stool samples were obtained from symptomatic 40 patients from outpatient clinic of Benha University Hospitals and Benha Educational Hospital. Twenty suffered from dysentery and 20 cases were not suffering dysentery. In addition 10 stool samples of other parasitic infection as Giardia lamblia and Cryptospordium parvum were examined by direct smear, iodine stained smear, formol ether concentration technique and real time PCR for detection of E. histolytica DNA. Formol ether concentration technique showed that 20 (40%) samples were positive for E. histolytica but real time PCR showed 26 (52%) positivity. All samples positive by microscopy were also positive by PCR and additional 6 PCR positive cases. There was no cross reaction with other parasites as G. lamblia and C. parvum. PMID- 21634246 TI - Babesiosis in an Egyptian boy aquired from pet dog, and a general review. AB - Human babesiosis has been documented in many countries. It is a zoonotic protozoan disease of medical, veterinary and economic importance. In this study, a twelve years old boy was referred to the hospital with intermittent fever of unknown origin. On clinical, parasitological and serological bases the case proved to be babesiosis. The boy acquired the infection from his pet dog which was heavily infested with Rhipicephalus sanguineus and suffered a mild feature of animal babesiosis. The patient was successfully treated with Atovaquone plus Azithromycin without relapse for one month follow up. The pet dog was sent to Governmental Veterinary Hospital at Abbassia for treatment from babesiosis and tick infestation. PMID- 21634247 TI - Geographical distribution and relative abundance of culicine mosquitoes in relation to transmission of lymphatic filariasis in El Menoufia Governorate, Egypt. AB - Culicine mosquito were surveyed in El Menoufia Governorate (October to November 2008 and April to May 2009) in villages representing eight districts. Six species were reported: Culex (Culex) pipiens Linnaeus, Cx. (Cx) perexiguus Theobald, Cx. (Cx.) antennatus (Becker), Aedes (Ochlerotatus) caspius (Pallas), Aedes (Ochlerotatus) detritus (Haliday) and Culiseta (Allotheobaldia) longiareolata (Macquart). Cx. pipiens, the main filariasis vector was the commonest or predominating species (ca. 47% adults & 92% larvae, P < 0.01). For the common species, the following were investigated: 1- temperature and pH of the breeding habitats and their relation to the larval density and 2- relation of adult indoor density with indoor- and outdoor- temperature and RH. Besides, parasitologically, Wuchereria bancrofti cases (33/631 blood samples, 5.23%) were detected in three districts (range = 1.96-14.12% infection). The cases were associated with the abundance of Cx. pipiens adults (ca. 45- 62% of the collected adults). PMID- 21634248 TI - Cholecysto cholangiography versus cystic duct cholangiography in laparoscopic cholecystectomy: a prospective controlled randomized trial. AB - This study evaluated the safety and effectiveness of cholecysto-cholangiography (CCC), an extremely rapid and easy way of imaging the biliary tree during laparoscopic cholecystectomy and a viable alternative to cystic duct cholangiography in the era of minimal invasive surgery. Sixty patients with symptomatic gallbladder stones were studied in this series, 30 were evaluated for cholecysto-cholangiography and other 30 for transcystic-cholangiography. The success rate in cholecysto-cholangiography approached 80% with excellent quality films obtained. Delineation of anatomy approached 85%; 75% for cholecysto cholangiography and transcystic cholangiography respectively. Exposure time to radiation compares favourably with cystic duct cannulation with a mean time 2.4 min. Cholecysto-cholangiography added less than 11 mins to the procedure; mean 10.4 mins, while transcystic cholangiography added an extra 30 mins; mean 31 mins. There were no cholangiogram related complications or false positive findings with a smooth for two months follow up. PMID- 21634249 TI - Head lice infestations (Anoplura: Pediculidae) in Saudi and non-Saudi school-aged children. AB - The prevalence and human risk factors associated with head lice infestation was studied. A total of 860 children, 474 (55.1%) boys and 386 (44.9%) girls, with ages ranged from 6-13 years were critically examined. They were randomly selected from two schoolchildren (one for boys and one for girls) in Jazan Province. The children or their parents were subjected to questionnaire and physical examination by visual inspections of scalp and hair for the presence of adult lice, nymphs, and/or eggs (nits). A total of 114 children (13.3%) were infested with head lice, 73 (18.9%) girls and 41 (8.6%) boys. The highest infestation rate was recorded in rural area (20.5%). Pediculosis had a negative association with father's education level and frequency of hair washing (p > 0.01). The results indicated that rural residence, sex, long hair, age- group 6- < 8 years old, positive history of previous infestation, sharing of beds and combs, uneducated mother and family overcrowding might be the risk factors for the head lice infestation. PMID- 21634250 TI - Carotid intima media thickness, left ventricular hypertrophy and weekly averaged blood pressure in hemodialysis patients. AB - Undoubtedly, cardiovascular complications are the leading cause of mortality and morbidity in haemodialysis (HD) patients, and hypertension plays an important role in development of cardiovascular disorders in them. The present study evaluated the weekly averaged blood pressure with its relation to carotid intima media thickness and left ventricular mass index in HD patients. The study included 112 HD patients (85 males and 27 females). We used daily home blood pressure (HBP) monitoring to record a total of 20 points of BP over a period of 1 week, including measurements of the wake-up and night BPs; in addition to the BP recorded before and after each HD session that occurred three times a week. The average of 20 BP measurements was defined as the weekly averaged blood pressure (WAB). Also, the relationship between WAB and left ventricular hypertrophy (LVH) or carotid intima media thickness and carotid intima media thickness and left ventricular hypertrophy were evaluated. The results showed that systolic WAB (144.26 +/- 7.39 mmHg) and diastolic WAB (.75.84 +/- 5.15 mmHg) were almost consistent with the wake-up BP on the day after the midweek dialysis session (R2 = 0.628 & 0.684, respectively). The WAB showed significant positive correlations with the left ventricular mass index (LVMI) (R = 0.387, P < 0.0003) and carotid intima media thickness (R = 0.226, P < 0.0034), whereas the predialysis systolic BP showed a significant positive correlation with the CIMT and non-significant correlation with LVMI. There was a significant positive correlation between CIMT and LVMI. PMID- 21634251 TI - Effect of mirazid (Commiphora molmol) on experimental giardiasis. AB - Mirazid -a purified oleo-resin extract derived from myrrh- with previously proved trematocidal activity, has been recently reported to be also effective against protozoa like Trichomonas vaginalis and cryptosporidium in humans. This experimental study aimed at investigating the possible therapeutic effect of mirazid on experimental Giardia lamblia infection in albino rats, using tinidazole as a therapeutic control. Results proved the efficacy of mirazid, as indicated by a 100% reduction in parasite- load of both intestinal and fecal parasitic counts, a direct toxic effect on Giardia trophozoite as revealed by SEM and TEM in form of cell membrane erosions together with increased cytoplasmic vaculation and dissociation, and also by reversal of the observed mucosal damage associated with Giardia after complete treatment. Starting trophozoite invasion of the mucosa was also demonstrated by TEM; its mechanism and possible pathogenic effect in giardiasis were discussed. PMID- 21634252 TI - Clinical, parasitological and social studies on Wuchereria bancrofti in Egypt. AB - This study was conducted on patients attending Al-Hussein University Hospital and Mansura University Hospitals. Sheets were filled out on each patient (age, residence, occupation, family similar condition, travelling, disease duration, symptoms suggesting acute dermatolymphangitis attacks, frequency and duration. They were 25 lymphoedema patients and 8 controls (17 males & 16 females) and subjected to clinical and parasitological examinations. According to residence, five patients were from Giza Govemorate (18.18%), four from Dakahlia G. (12.12%), four from Gharbia G. (12.12%), five from Menoufia G. (15.15%), four from Sharkia G. (12.12%), two cases from Kafr Elsheikh G. (6.06%) and one patient from Assiut G. (3.03%). In controls, 6 were from Al-Hussein Cairo G. (15.15%), and two cases (6.06) from Al-Abbassia. They were 48% farmers (10 male & 2 female); 4% grocers (1 male); 8% carpenters (2 male); and 40% house wife (10 female). According to lymphedema site: 3 cases (12%) were bilateral lower limbs lymphedema (2 female & 1 male), 13 cases (52%) had right lower limb lymphedema (6 female & 7 male), and 9 cases (36%) had left lower limb lymphedema (4 female & 5 male). They were 18 (72%) with below knee extension of lymphedema (10 male & 8 female) and 7 (28%) above knee extension of lymphedema (3 male & 4 female). Grade of lymphedema was one case (male) with grade II lymphedema (4%), 13 cases (52%) with grade III (7 male & 6 female) and 11 cases (44%) with grade IV (5 male & 6 female). Duration of lymphedema was one case with grade II lymphedema in a period of 3 years, 13 cases with grade III lymphedema and the durations of lymphedema for these cases were (one case with 5 years duration, 3 cases were > 6-9 years,6 cases were > 9 12 years and 3 cases were > 12 years) and 11 cases with grade IV lymphedema and duration of lymphedema for these cases were (one case with 6 years duration, 2 cases were > 6-9 years,3 cases were > 9-12 years and 5 cases were > 12 years). PMID- 21634253 TI - The current status of fleas according to environmental changes in some Governorates in Egypt. AB - A preliminary survey of domestic rodent and their fleas was carried out in different environmental Governorates (Ismailia, Dakahlia, El-Fayoum, North Sinai and Matrouh), Egypt. Flea index (number of flea/rodent) and percentage frequency of different flea species were recorded in spring (2009- 2010). The main rodent species found were the Norway rat, Rattus norvegicus, the grey-bellied rat, Rattus rattus alexandrinus, the white- bellied rat, Rattus rattus frugivorus, the house mouse, Mus musculus and the spiny mouse Acomys cahirinus. The common flea species attacking rodents were: the oriental rat flea, Xenopsylla cheopis, the mouse flea, Leptopsylla segnis, the dog flea, Ctenocephalides canis and the sticktight flea Echidnophaga gallinacea. The flea index at Ismailia and Matrouh Governorates showed the highest indices (8.93 & 7.68), while El-Fayoum and North Sinai Governorates showed the lowest ones (1.09 & 1.68). Dakhalia showed moderate flea index (4.52). The highest number of fleas was recorded on R. norvegicus lives in places that are easy to dig barrows, which are suitable medium for fleas breeding. The lowest number was recorded on Mus musculus and Acomys cahirinus. The oriental rat flea, Xenopsylla cheopis was the highest frequency distribution for all domestic rodent species, while, the stick-tight flea, Echidnophaga gallinacea was the lowest which recorded at Ismailia and Dakahlia only. PMID- 21634254 TI - Echinochasmus aspinuosa as a new record species in Egyptian heron. AB - This paper reported a new Egyptian of Echinochasmus aspinuosa in herons trapped in Giza and Sharkia Governorates. The detailed morphology was given and illustrated. The zoonotic importance of this digenetic treamatode was discussed. PMID- 21634255 TI - Clonorchis sinensis a new report in Egyptian employees returning back from Saudi Arabia. AB - Clonorchis sinensis, or Chinese liver fluke, is a parasite of the fish-eating mammals endemic in the East Asia, Far East, particularly in China, Japan, Taiwan, Vietnam, and Korea and among immigrant refugees to European Union and Eastern Mediterranean Countries. This paper reported infection among Egyptian family who practice consumption of imported fishes from the Far East. Diagnosed was based on detection of its characteristic eggs, positive specific ELISA, radiological and family behavior and feeding habit history. Patients were successfully treated. PMID- 21634256 TI - Coexistence of two species of haplosporidian parasites in a population of the marine amphipod Parhyale hawaiensis with evidence for parasite phagocytosis and transmission mode. AB - The amphipod Parhyale hawaiensis live hiding underneath stones and shingles along the shore line in supra-littoral zone of Sharm El-Nagha site, Hurghada city, Red Sea. The collected specimens of P. hawaiensis are infected with two protozoan parasites, Urosporidium sp. and Haplosporidium sp. which invade many organs of both females and males. The life cycle of the parasites was described in addition to their histopathological effects on host's tissues. The study showed some evidence for the defense reaction of the host as engulfing and degraded stages of parasites by granulocytes. The two parasite species coexist at different prevalence's in one host population and bi-infected individuals were few. PMID- 21634257 TI - Nosocomial infections and infection control process in Egypt. PMID- 21634258 TI - Of private and public safety nets. PMID- 21634259 TI - A much better health care system. PMID- 21634260 TI - Choice in public health insurance: evidence from West Virginia medicaid redesign. AB - This paper examines the factors that affect plan choice in a public health insurance program. West Virginia recently redesigned its state Medicaid program, offering members a choice between two plans--a basic plan and an enhanced plan. The latter plan includes more benefits, but requires additional agreements intended to lead patients to adopt healthier lifestyles. We use administrative claims records and survey data to examine plan choice. Our results yield convincing evidence that members with higher health care utilization patterns are more likely to enroll in the enhanced plan, but other factors such as education are also important. PMID- 21634261 TI - The impact of private long-term care insurance on the use of long-term care. AB - This paper investigates the effects of privately purchased long-term care insurance (LTCI) on three major types of long-term care services: nursing home care, paid home care, and informal care received from Family and friends. Using 2002-2008 data from the ongoing Health and Retirement Study, we analyze the determinants of long-term care utilization simultaneously with the determinants of holding LTCI. We find that LTCI has modest effects on the likelihood of using long-term care services. For the very frail elderly, private LTCI enhances their access to nursing home care. For those with moderate disability, LTCI makes it more likely that they can remain at home and receive home care services, instead of going to a nursing home. We find no evidence that formal care substitutes for informal care in the presence of LTCI. These findings suggest that if LTCI becomes much more prevalent in the future, many older adults will be able to choose the type of long-term care arrangement that best suits their needs. PMID- 21634262 TI - How does health insurance affect the retirement behavior of women? AB - The availability of health insurance is a crucial factor in the retirement decision. Women are substantially less likely to have health insurance from their own employment. Using the Health and Retirement Study, we examine the role of employer-provided retiree health insurance in the retirement decisions of single women, and women in single-earner and dual-earner couples. We compare the effect of health insurance on female and male retirement. Our results show that retiree health insurance increases retirement for all groups except single men. We find suggestive evidence that the role of health insurance for women hinges on their husbands' labor force status. PMID- 21634263 TI - A new approach to reducing payments made to hospitals with high complication rates. AB - This article proposes a redesign of the Medicare inpatient prospective payment system to reduce payments made to hospitals with high complication rates. We compute risk-adjusted, expected complication rates for hospitals and compare them to actual complication rates in order to determine the number of excess complications. Hospital payment reductions then are computed based on the number of excess complications in a hospital. Medicare hospital payment could be reduced by approximately 8% (8.5 billion dollars) if hospitals were held to a "best practice" standard and if payments made for excess complications were eliminated. PMID- 21634264 TI - Gordon T. Edwards lessons learned about EHR incentive payments. PMID- 21634265 TI - ACO regs, round 1. PMID- 21634266 TI - Future pension accounting changes: implications for hospitals. AB - Proposed rules in accounting for defined benefit plans may affect hospitals' statement of operations and affect the time, effort, and cost to comply with periodic financial reporting requirements. The new standard would require immediate recognition of the full amount of plan amendments in determining operating income. Hospitals should consider the role of pension plans in their compensation programs. PMID- 21634267 TI - Strengthening revenue cycle capabilities in an era of reform. AB - Strategies that healthcare finance professionals should incorporate to help their organizations respond effectively to payment reforms include: Assessing the organization's ability to capture and share relevant data. Educating themselves, the board of trustees, and the medical staff on pertinent rules as payment reforms are rolled out. Examining inefficiencies related to care processes. Establishing policies and procedures to address "commingled" data. PMID- 21634268 TI - Nicholas Wolter putting patients first under payment reform. PMID- 21634269 TI - A new road map for healthcare business success. AB - Action steps hospitals should take to prepare their organizations for a changing business development environment include: Developing a comprehensive forecast of the ways in which reform and market forces will affect patient volumes and service line demand. Aligning with physicians and other care entities in a tightly integrated way. Heightening transparency related to quality and cost. Investing in marketing and social media to strengthen the organization's market position. PMID- 21634270 TI - Can a hospital benefit from partnering with physicians? AB - The prospect of accountable care organizations gives hospitals good reason today to consider the benefits of developing an ambulatory surgery center (ASC) as a joint venture with physicians, as well as the risks of not pursuing such a strategy. A key potential benefit that such an ASC can offer a hospital is the ability to enhance a hospital's physician recruitment efforts and increase surgical volume. Another important consideration is whether the hospital and physicians possess the collective expertise, performance track record, capital, tenacity, perseverance, resources, and credibility to perform the necessary functions of the ASC on their own. PMID- 21634271 TI - 7 strategies for success in governing an ACO. AB - Strategies for success in governing an ACO include: Clearly articulating a patient-centered strategic plan that can serve as the basis for ACO management. Capitalizing upon each partner's strengths. Ensuring that capital and resources are allocated consistently with the ACO's goals and objectives. Establishing effective leadership and engaging board members in governance. Developing legal and organizational structures that will best facilitate an integrated model. Employing the right technology in a way that best supports the ACO's mission. PMID- 21634272 TI - Uncovering pharmacy department risks and opportunities. AB - To assess the risk a hospital faces from improper billing, coding, and pricing for pharmacy items, hospital finance leaders should perform an audit of the pharmacy department's charge description master. The audit should look for inaccuracies with respect to: National drug codes. Healthcare Common Procedure Coding System codes. UB-04 revenue codes. Billable units. Wholesale acquisition costs and average wholesale prices. PMID- 21634273 TI - Success under reform through revenue cycle excellence. AB - Hospitals and health systems should take the following steps to improve their revenue cycle performance: Collect patient responsibility amounts up front. Reduce credit balance accounts. Reduce preregistered patient no-shows. Identify and manage unbilled accounts receivable. PMID- 21634274 TI - Healthcare BI: a tool for meaningful analysis. AB - Implementing an effective business intelligence (BI) system requires organizationwide preparation and education to allow for meaningful analysis of information. Hospital executives should take steps to ensure that: Staff entering data are proficient in how the data are to be used for decision making, and integration is based on clean data from primary sources of entry. Managers have the business acumen required for effective data analysis. Decision makers understand how multidimensional BI offers new ways of analysis that represent significant improvements over historical approaches using static reporting. PMID- 21634275 TI - The impact of healthcare reform on hospital costing systems. AB - Hospitals should consider five approaches to costing over the next three years as they prepare for reform: Moving from a strategic to an operational cost-finding model. Adopting job costing over standards costing whenever possible. Reviewing working classifications of cost with managers and administrators. Evaluating current cost bucketing definitions and reviewing the expenses assigned to these buckets. Increasing the use of microcosting. PMID- 21634276 TI - Cloud computing: innovating the business of health care. PMID- 21634277 TI - Endowment income for the post-reform era. PMID- 21634278 TI - The evolution of physician compensation: what's next? PMID- 21634279 TI - Attack and defend. A primer on preventing and spotting strokes. PMID- 21634280 TI - Vintage advice. The art of appreciating good wine. PMID- 21634281 TI - Race of a life time. Nat Strand's amazing ways. PMID- 21634282 TI - Dreaming of a healthy tomorrow. New evidence about the role of sleep in type 2 diabetes. PMID- 21634283 TI - "Kids, I have diabetes". How to talk to children about your disease. PMID- 21634284 TI - Treat yourself right. A guide to cooking for one. PMID- 21634285 TI - The EEOC's regulations implementing the ADAAA: expanding coverage under the "disability" definition. PMID- 21634286 TI - The experience of Stinner, et al. PMID- 21634287 TI - RE: tobacco product usage in Afghanistan: deployed military personnel versus deployed civilian contractors. PMID- 21634288 TI - Global health diplomacy: a call to action. PMID- 21634289 TI - War poets and the ageless anger of Achilles. PMID- 21634290 TI - Starting a primary care pain management clinic in a military treatment facility. PMID- 21634291 TI - The interaction of body armor, low-intensity exercise, and hot-humid conditions on physiological strain and cognitive function. AB - OBJECTIVE: This project was aimed at evaluating the impact of combat armor on physiological and cognitive functions during low-intensity exercise in hot-humid conditions (36 degrees C and 60% relative humidity). METHODS: Nine males participated in three trials (2.5 hours), walking at two speeds and wearing different protective equipment: control (combat uniform and cloth hat); torso armor with uniform and cloth hat; and full armor (uniform, torso armor, and helmet). RESULTS: As time progressed, core temperatures increased and deviated significantly among trials, rising at 0.37 degrees C h(-1) (control), 0.41 degrees C h(-1) (torso armor), and 0.51 degrees C h(-1) (full armor). Heart rates also progressively diverged, and subjects lost significantly more sweat during the two armored trials. However, cognitive-function tests revealed neither significant main effects nor time by treatment interactions. CONCLUSION: The combat armor and helmet significantly increased thermal and cardiovascular strain, but these were unlikely to lead to either exertional heat illness or impaired cognitive function during uneventful urban, military patrols in hot humid conditions. PMID- 21634292 TI - Energy expenditure estimation during daily military routine with body-fixed sensors. AB - The purpose of this study was to develop and validate an algorithm for estimating energy expenditure during the daily military routine on the basis of data collected using body-fixed sensors. First, 8 volunteers completed isolated physical activities according to an established protocol, and the resulting data were used to develop activity-class-specific multiple linear regressions for physical activity energy expenditure on the basis of hip acceleration, heart rate, and body mass as independent variables. Second, the validity of these linear regressions was tested during the daily military routine using indirect calorimetry (n = 12). Volunteers' mean estimated energy expenditure did not significantly differ from the energy expenditure measured with indirect calorimetry (p = 0.898, 95% confidence interval = -1.97 to 1.75 kJ/min). We conclude that the developed activity-class-specific multiple linear regressions applied to the acceleration and heart rate data allow estimation of energy expenditure in 1-minute intervals during daily military routine, with accuracy equal to indirect calorimetry. PMID- 21634293 TI - Weight changes among male Navy personnel deployed to Iraq or Kuwait in 2005-2008. AB - The objective of this study was to identify changes in weight that occurred during deployment to Iraq or Kuwait between 2005 and 2008. Data on length and type of deployment among 16,365 male U.S. Navy personnel were combined with weight measurements before and after deployment from the Physical Readiness Information Management System. Weight measurements were available for 10,886 men who did not exceed Navy weight recommendations before deployment. In general, weight increased after deployment and, for those who did not exceed Navy recommendations before deployment, factors associated with weight gain included being enlisted and having a deployment longer than 228 days. Among 1,108 men with 2 deployments, a dwell time shorter than the combined deployed time was a risk factor for weight gain during the second deployment. Future studies should explore the combined effects of long deployments and short dwell times in maintaining the readiness of military personnel. PMID- 21634294 TI - Obesity in the UK Armed Forces: risk factors. AB - OBJECTIVE: To assess the risk factors of obesity in terms of body mass index (BMI) and waist circumference in the UK Armed Forces. METHODS: A quota sample from all UK services, of 2,448 men and 311 women aged 17-55 years. RESULTS: The risk ratios for those with a BMI 227.5 kg/m2 and waist circumference 294 cm in men were 0.21 (95% confidence interval [CI], 0.13-0.34) for non-whites compared to whites, 0.55 (95% CI, 0.38-0.78) for commissioned officers compared to lower ranks, and 0.13 (95% CI, 0.06-0.31) for Royal Marines compared to the Army, but increased with age, 58.2 (95% CI, 19.6-172.7). Risk ratios were usually nonsignificant between service-demographic factors for BMI > or = 27.5 kg/m2 and waist circumference < 94 cm. CONCLUSIONS: The risk of obesity is highest in the Army and Royal Navy, whites, lower ranks, and older ages. BMI and waist circumference should be used together as measures of obesity. PMID- 21634295 TI - Validity and reliability of the 20 meter shuttle run test in military personnel. AB - is regularly monitored in military personnel, as occupational demands require a certain level of fitness. Distance run (eg, 2 mile) is typically carried out to measure aerobic fitness, but an alternative test could be the 20 meter shuttle run test (20 m SRT). The present study aimed to evaluate validity and reliability of this test in military personnel. An equation for predicting maximal oxygen uptake (VO2max) was developed on 38 Home Guard soldiers and cross validated in 28 Air Force cadets. Reliability of the 20 m SRT, expressed as mean difference in estimated VO2max-- 95% limits of agreement, was -0.8 +/- 3.1 mL x kg(-1) min(-1). Mean difference +/- limits of agreement between estimated and measured VO2max was -0.4 +/- 6.2 mL.kg(-1)x min-'. The 20 m SRT seems to be a reliable test, although validity is less certain, as relatively high variability was observed between measured and estimated VO2max from the 20 m SRT. PMID- 21634296 TI - U.S. military recruits waived for pathological curvature of the spine: increased risk of discharge from service. AB - Selective accession waivers for medically disqualifying conditions like spinal curvature are one way the military meets its manpower needs. We evaluated retention patterns during the first 2 years of service of a cohort of military recruits with waivers for pathological curvature of the spine (spinal curvature). Recruits waived for spinal curvature (n = 417), who accessed from 1998 to 2005 were identified and matched with 3 qualified recruits. Kaplan-Meier survival analysis and Cox proportional hazards model were used to compare survival patterns and adjusted attrition hazard estimates. Waived recruits experienced significantly increased risk of "all cause" discharge (relative risk = 1.3; 95% confidence interval: 1.1, 1.5) and "existing prior to service" discharge (relative risk = 2.4; confidence interval: 1.6, 3.5). Despite the increased risk of discharge, current waiver criteria allowed a majority with spinal curvature to complete at least 2 years of service. Policy makers must consider risks and benefits before modifying the current accession standard for spinal curvature. PMID- 21634297 TI - Incidence and predictors of mental health hospitalizations in a cohort of young U.S. Navy women. AB - PURPOSE: This prospective cohort study analyzed hospitalization data over a 7 year period for 5,503 female Navy recruits first assessed in 1996-1997. METHODS: Participants' age, education, race/ethnicity, occupation, and traditionality of military occupation were analyzed as predictors of hospitalization for mental health diagnoses occurring 7-8 years after entry into the Navy. RESULTS: 5.2% of women was hospitalized at least once for a mental health disorder over the 7-8 year follow-up period. Thirteen percent of all hospitalizations for the cohort were for mental health reasons. Adjustment reaction/acute stress reaction and personality disorders were the 2 most frequent mental disorders. Asian/ Pacific Islanders with more than a high school education and those in the younger age ranges were significantly less likely than others to be hospitalized for a mental disorder. CONCLUSIONS: Results underscore that mental health issues among young military women during peacetime are not a trivial source of morbidity. PMID- 21634298 TI - Felo De Se: soldier suicides in America's Civil War. AB - OBJECTIVE: This article examined the factors associated with suicide during America's Civil War and the years immediately following the cessation of armed conflict. METHODS: Contemporary newspaper reports, complemented by book and journal articles, provide an understanding of the incidence and motivations of suicide. RESULTS: The rate of suicide in the general population dramatically increased in the years following the war's end. During the Civil War, suicides occurred nearly every month, reliably peaking in the spring of each year. Depression and alcohol abuse were major factors associated with military suicides. CONCLUSION: Emotional disorders and alcohol misuse, when combined with the hardships of war, contributed to a steady rate of suicides during the Civil War. PMID- 21634299 TI - Descriptive epidemiology and natural history of idiopathic venous thromboembolism in U.S. active duty enlisted personnel, 1998-2007. AB - The estimated incidence of idiopathic venous thromboembolism (IVTE) cases in the United States ranges from 24,000 to 282,000/year. This analysis explores the incidence and prevalence of IVTE in the military and if cases experience increased attrition. The Defense Medical Surveillance System was searched for incident IVTE cases from 1998 through 2007. Enlisted cases were each matched to 3 controls. Kaplan-Meier survival analysis and Cox proportional hazard modeling were performed. We matched 463 cases to 1,389 controls. Outpatient IVTE rates have increased markedly from 1998 through 2007. Cases of all-cause attrition risk (0.56 [95% CI = 0.44, 0.72]) and rates were significantly less than controls (p < 0.0001), and cases of medical attrition risk (1.64 [95% CI = 1.13, 2.37]) and rates were significantly higher (p < 0.01). Increasing rates with lower attrition suggests increasing case prevalence. Health care providers must maintain a high index of suspicion for venous thromboembolism to minimize adverse sequelae affecting health, unit readiness, and medical costs. PMID- 21634300 TI - Low back pain in the uniformed service member: approach to surgical treatment based on a review of the literature. AB - The clinical entity of chronic low back pain is not well understood at present. Disparate spinal conditions can result in symptoms of low back pain, and various treatment options have been proposed in the literature including surgical intervention and prolonged nonoperative therapy. Low back pain, as well as lumbar degenerative disc disorders and spondylosis, is common among uniformed service members, and studies, along with treatment recommendations, regarding these entities in the civilian community are not necessarily translatable to military personnel. This literature review endeavors to review the evidence for interventions for low back pain and seeks to formulate a treatment algorithm that takes into account the unique requirements of military personnel. Ultimately, more research must be done, investigating the specific results of treatments for spinal conditions within the military community before definitive recommendations can be made. PMID- 21634301 TI - High prevalence of chronic obstructive pulmonary disease among veterans in the urban midwest. AB - Although chronic obstructive pulmonary disease (COPD) is a leading cause of morbidity and mortality within the Veterans Health care Administration, its prevalence and recognition are not known. We measured airflow limitation and diagnosed COPD at the Cincinnati Veteran's Administration Medical Center. Participants were 326 outpatients who performed spirometry and completed questionnaires. Health care-provider-diagnosis and self-diagnosis of COPD were compared with COPD defined by forced expiratory volume in 1 second (FEV1)/forced vital capacity (FVC) < 0.7 (fixed ratio) and (FEV1/FVC)/lower limit of normal (LLN) < 1.0. COPD prevalence was 43% (95% confidence interval: 36.9, 48.1) by fixed ratio and 33% (95% confidence interval: 27.2, 36.8) by LLN. Eighteen percent of the patients had health care-provider-recorded and 23% had self reported diagnoses of COPD. Positive predictive values for the diagnosis of COPD were 79% and 64% for healthcare providers versus 68% and 62% for patients; negative predictive values were 64% and 74% for healthcare providers versus 64% and 76% for patients (fixed ratio and LLN, respectively). COPD prevalence is higher among Cincinnati veterans than among general U.S. population. COPD is under-recognized by both health care providers and veterans. PMID- 21634302 TI - Prevalence of snoring and sleep-disordered breathing among military personnel. AB - OBJECTIVE: The prevalence of snoring as well as signs and symptoms suggestive of sleep-disordered breathing (SDB) and obstructive sleep apnea syndrome (OSAS) among a military population was investigated. METHODS: The study was a questionnaire-based survey carried out over military personnel obtained in 3 randomly selected typical military bases in the United Kingdom. Data analysis used contingency table methods and logistic regression modeling. RESULTS: Regular snoring was significantly associated with male sex, breathing pauses during sleep, not waking up refreshed in the morning, ingestion of sleeping tablets and state of health; 19.5% of the subjects snored regularly. Based on the minimum criteria of the International Classification of Sleep Disorders and the questionnaire results, 2.9% (38 subjects) presented symptoms suggestive of SDB or OSAS. CONCLUSION: With the possible implications for the military, greater attention with regards to SDB or OSAS is required, especially to its recognition and management. PMID- 21634303 TI - Effects of 6 weeks of quercetin supplementation on energy, fatigue, and sleep in ROTC cadets. AB - PURPOSE: To investigate the effects of 6 weeks of quercetin supplementation on energy, fatigue, and sleep quality in young persons conducting military physical training. METHODS: Using a randomized, double-blind, repeated-measures, placebo controlled design, 58 healthy, moderately trained men and women were assigned to quercetin group and placebo group. Energy, fatigue, and sleep quality were evaluated before, in the middle, at the end, and 2 weeks following 42 to 54 days of supplementation with 1 g/day of quercetin in a soft chew or a placebo. RESULTS: Changes in energy and fatigue, assessed by the Profile of Mood States Brief and the Mental and Physical State Energy and Fatigue Scales, and changes in sleep quality, measured by the Pittsburgh Sleep Quality Index were not significantly different (p > 0.05) in quercetin and placebo groups. CONCLUSION: Six weeks of quercetin supplementation in young persons conducting military physical training does not influence energy, fatigue, or sleep quality. PMID- 21634304 TI - Colonoscopy by a family physician: detecting proximal neoplasia in asymptomatic adults. AB - PURPOSE: Flexible sigmoidoscopy (FS) is considered an adequate screening test in average risk (AR) patients. The purpose of this study was to determine the diagnostic yield of FS in detecting colonic neoplasia between AR and high risk (HR) patients. METHODS: We present a chart review of 559 outpatient colonoscopies performed by a family physician from September 2003 to October 2007. The prevalence of neoplasia and diagnostic yield of FS was compared between groups. RESULTS: The overall prevalence of neoplasia was 23.1% (AR) and 32.8% (HR); p = 0.02. The prevalence of proximal neoplasia not detectable by FS was 10.2% (AR) and 14.5% (HR); p = 0.16. The diagnostic yield of FS in each group was 56%; FS would have missed 44% of polyps regardless of patient risk. CONCLUSIONS: The high rate of colonic neoplasia not detected by FS (44%) suggests that FS alone is inadequate for screening in AR patients. PMID- 21634305 TI - Continuous thoracic paravertebral nerve block in a working anesthesia resident when opioids are not an option. AB - Multiple unilateral rib fractures can cause significant pain and morbidity. Continuous nerve block catheters are often maintained while inpatient, and patients are discharged with oral analgesics. However, in many institutions, this dynamic is changing and patients are being managed effectively with outpatient catheters. A 45-year-old male was presented with fractured right ribs 6 through 9. The patient was an anesthesiology resident and was unable to perform his clinical duties. Single paravertebral nerve blocks were performed at right thoracic levels 6-9. At the T7 level, an indwelling catheter was placed. On post injury day 18, he was able to discontinue the catheter and there were no associated complications. We report a unique case of a patient with multiple rib fractures who was not able to be exposed to potential side effects of opioids. The use of a continuous thoracic paravertebral nerve in an outpatient setting allowed a faster return to function with no adverse events. PMID- 21634306 TI - Leucocytoclastic vasculitis in severe ulcerative colitis. AB - Ulcerative colitis may be associated with a number of extraintestinal skin manifestations including erythema nodosum and pyoderma gangrenosum. We describe an unusual case of a 26-year-old military pilot with ulcerative colitis and skin lesions diagnosed as leucocytoclastic vasculitis. The skin lesions occurred twice during the severe flare. The first occurrence was treated successfully with corticosteroids. When the lesions recurred several weeks later, concomitant with the ongoing flare, treatment with mesalamine and infliximab resolved the lesions. Ultimately, the patient required total colectomy for flare unresponsive to maximum medical therapy. The leucocytoclastic vasculitis did not recur after colectomy. We propose a potential immunopathophysiologic mechanism linking the 2 conditions based on recent biochemical and clinical research. PMID- 21634307 TI - Permanent press allergy in an active duty U.S. Army soldier. AB - Ethylene urea/melamine formaldehyde resin (permanent press) is a common fabric finishing agent added to Army Combat Uniforms for a wrinkle-free appearance and to strengthen the fabric. We describe the case of an active duty U.S. Army soldier with a diffuse eczematous dermatitis in whom patch testing was used to identify an allergy to permanent press, a ubiquitous fabric finishing agent in the Army combat uniform. To our knowledge, this is the first case report of a soldier with an allergic contact dermatitis to ethylene urea/melamine formaldehyde resin. This case highlights the importance of considering the diagnosis of allergic contact dermatitis in patients with a recurrent eczematous dermatitis that does not respond appropriately to therapy and the unique occupational impact of diagnosing an Army soldier with permanent press allergy. PMID- 21634308 TI - Disseminated nocardia cyriacigeorgica presenting as right lung abscess and skin nodule. AB - Nocardia species are ubiquitous in the environment and can be found worldwide. Direct inhalation remains the most commonly attributed route of infection with Nocardia asteroides complex, causing 50% of invasive infections. Improved molecular methods have identified a significant proportion of N. asteroides complex isolates to be Nocardia cyriacigeorgica. We report a case of a 58-year old male working as a contractor in Bagram Air Force Base, Afghanistan, with disseminated N. cyriacigeorgica involving the lung, brain, and dermis. Diagnosis was facilitated by early identification of branched, filamentous bacteria using Fite and gram staining along secA DNA sequencing of clinical isolates. Our patient is the first confirmed case of N. cyriacigeorgica infection in Afghanistan. PMID- 21634309 TI - Sentinel case of group A beta-hemolytic streptococcus causing constrictive pericarditis presenting as hypogammaglobulinemia. AB - This is a unique case of a previously healthy 7-year-old boy, which highlights the importance of considering immunodeficiency when a rare infection occurs. In the following case report, the patient develops constrictive pericarditis secondary to group A beta-hemolytic streptococcal infection. As a result of this infection, we speculate that he develops hypogammaglobulinemia secondary to the documented association between constrictive pericarditis and intestinal lymphangiectasia because an extensive work-up for a primary immunodeficiency was negative. This is the first case ever to present constrictive pericarditis because of group A beta-hemolytic streptococcal infection. PMID- 21634310 TI - A pedometer-based intervention to improve physical activity, fitness, and coronary heart disease risk in National Guard personnel. AB - To compare the effects of a pedometer-based behavioral intervention (Fitness for Life [FFL] program) and a traditional high-intensity fitness (TRAD) program on physical activity (PA), Army Physical Fitness Test (APFT), and coronary heart disease risk factors in Army National Guard members who failed the APFT 2-mile run. From a pool of 261 Army National Guard, a total of 156 were randomized to TRAD or FFL for 24 weeks consisting of a 12-week progressive conditioning program followed by 12 weeks of maintenance. For both groups, the total APFT score and 2 mile run time/score improved from baseline to 12 weeks (FFL: down 7.4%, p = 0.03; TRAD: down 5%, p = 0.08) but at 24 weeks they had regressed toward baseline. PA improved modestly and coronary risk profile changed minimally in both groups. A pedometer-based exercise intervention had results similar to a high-intensity program for improving PA, APFT, and 2-mile run times/score. Neither group sustained the improved run times over the 12 weeks of maintenance. PMID- 21634311 TI - Disaster strikes! Long-term care resident outcomes following a natural disaster. AB - This report describes the outcomes of 17 long-term care residents who were participating in a nursing intervention study. The residents were evacuated for 5 days due to a severe summer storm that caused widespread power outages. These residents were seen the day of the storm and three times per week for 2 weeks following their return to the nursing home. More than half of the participants had significant changes in their NEECHAM Confusion Scale scores (n = 11) and modified Confusion Assessment Method scores (n = 9) scores, suggesting the onset of delirium. Two participants were hospitalized within the 2 weeks of the evacuation. One participant died unexpectedly. This report provides a rare look into the negative effects of a short-term evacuation due to a natural disaster. PMID- 21634312 TI - Lumbar spinal stenosis. Clinical considerations for older adults. AB - Lumbar spinal stenosis (LSS) is becoming more frequent as the population ages and is now the most common spinal diagnosis for individuals older than 65. Because LSS is a common source of pain and disability among older adults, understanding the pathophysiology, clinical presentation, and clinical management of this condition is important. An individual example is used to highlight classic signs and symptoms. Nursing strategies for LSS-associated pain and symptom management to improve physical function and quality of life are discussed. PMID- 21634313 TI - Reassessing nurse aide job satisfaction in a Texas nursing home. AB - This article reports a study that replicates and extends Castle's 2007 study by examining factors related to satisfaction of nurse aides at Carillon House, a 120 bed nonprofit skilled nursing facility in Lubbock, Texas. The Nursing Home Nurse Aide Job Satisfaction Questionnaire was adapted to allow for the collection of qualitative responses and administered to the nursing staff. The results suggest that satisfaction among nurse aides is related to rewards, workload, and the team environment created among coworkers. These findings differ from what is generally found in the literature and may be related to the higher-than-average satisfaction rating of nurse aides at this facility. The study provides evidence that large-scale surveys may have ignored a stratified effect where higher satisfaction organizations have different driving forces than what has been demonstrated in the literature to date. PMID- 21634314 TI - Health literacy among older adults: a systematic literature review. AB - The purpose of this systematic literature review was to describe the empirical findings on health literacy in adults 65 and older. A systematic search for research studies from 1999 to early 2010 was conducted in MEDLINE and CINAHL databases. Articles were analyzed using the following foci: skills and health knowledge, functional health literacy, and health behaviors and health outcomes. Findings indicated that advancing age resulted in a significant increase in prevalence of inadequate health literacy. Further research is needed to develop evidence-based, culturally congruent interventions to improve health literacy among older adults. PMID- 21634315 TI - Recurrent falls among community-dwelling older Koreans: prevalence and multivariate risk factors. AB - This study was conducted to determine the risk factors for nonfallers versus fallers (1+ falls) and nonfallers/one-time fallers versus recurrent fallers (2+ falls) using the Resident Assessment Instrument-Home Care (RAI-HC((c))). Community-dwelling Koreans 65 and older (N = 411) comprised the sample. Multivariate logistic regression was used to determine the factors predictive of fallers and recurrent fallers. Eight predictive factors were statistically significant with regard to recurrent falls: unsteady gait, low scores related to performance of activities of daily living (ADLs) and instrumental ADLs (IADLs), high pain scores, number of housing environmental hazards, use of an assistive device, fear of falling, and reduced vision. Based on the findings, it is important to assess the risk factors for recurrent falls and develop differentiation strategies that help prevent recurrent falls, including management of gait problems, pain control, use of appropriate assistive devices, a fear management program, regular eye examinations, making improvements to ADLs and IADLs, and creating a safer home environment. In addition, using a standardized tool such as the RAI-HC would help assess multivariate fall risk factors to facilitate comparisons across different community care settings. PMID- 21634316 TI - Stakeholder opinions on a transformational model of pain management in long-term care. AB - Pain in older adults with dementia who reside in long-term care (LTC) facilities tends to be undertreated, despite important guidelines designed to ameliorate this problem. A group of public policy and geriatric pain experts recently concluded that existing guidelines are not being implemented because they fail to take into account policy and resource realities. The group published a set of more feasible guidelines that confront these realities (e.g., a recommendation for very brief pain assessments that can be conducted by nursing staff at least weekly). We asked stakeholders to provide opinions on the possibility of implementation of these guidelines within their LTC facilities. Our results support the feasibility of, interest in, and desirability of implementation. They also support an increased role for nurse leadership in LTC pain management. These results could be used to strengthen advocacy efforts for improvement in pain management. PMID- 21634317 TI - Barriers to depression care for black older adults. Practice and policy implication. AB - Late-life depression is a public health problem in the United States. Untreated, depression contributes to poorer health outcomes and increased mortality among older adults. Specifically, Black older adults are at higher risk for misdiagnosis, undertreatment, and more severe depressive symptomatology than other groups. Barriers to identification and treatment of depression in Black older adults include lack of access to quality mental health care, the stigma of mental illness, mistrust of mental health providers, and poor provider-client communication. Recommendations for gerontological nursing practice, education, and research to improve the care of depressed Black older adults are discussed. Implications for policy development are presented. PMID- 21634318 TI - Family matters. Family quality of life in Dementia. PMID- 21634322 TI - The Institute of Medicine report: education's role in serving the public's interests. PMID- 21634323 TI - Nursing for the future, from the past: two reports on nursing from the Institute of Medicine. AB - In 2010, the Institute of Medicine, in association with the Robert Wood Johnson Foundation, released a major study of the nursing profession. The Institute of Medicine last published a major, broad-based study of the nursing profession in 1983. This brief historical analysis examines the context and key recommendations for each report and provides concrete examples of outcomes of the 1983 report. We argue that despite similarities in context and recommendations, the two reports differ in target audience and implementation strategies, and that nursing is currently better positioned to use the report as a blueprint to improve patient care, as well as garner outside support, than it was in 1983. PMID- 21634324 TI - Interprofessional education: an essential strategy to accomplish the future of nursing goals. AB - The Institute of Medicine's Future of Nursing report has called for broad changes in the nursing profession for the benefit of society. Interprofessional education is one strategy to improve nursing education and enhance the role of nurses as collaborative leaders in the health care system. These are important lessons that have been learned from recent interprofessional educational initiatives and some encouraging examples of successful initiatives that are breaking down barriers across professional schools. The ultimate beneficiary of these changes will be our patients, who will receive better care. PMID- 21634325 TI - Through a glass darkly: reflections on change in health care, nursing, and blue ribbon committees. AB - The confluence of the Future of Nursing report and the calls for comprehensive reform of the health care system in the United States presents a unique opportunity for nursing to redefine its leadership role. This redefinition should be mindful of those places where nursing has and does provide leadership and the unique set of competencies that every nurse possesses that can be focused on leadership work. PMID- 21634326 TI - Preparing nurses for participation in and leadership of continual improvement. PMID- 21634327 TI - Engaging students using feature films. PMID- 21634360 TI - IGF-I and visfatin levels in retinopathy of prematurity. AB - PURPOSE: The authors hypothesized that circulating visfatin concentrations in preterm infants might be linked with retinopathy of prematurity (ROP)-like insulin-like growth factor I (IGF-I) levels. METHODS: The study group comprised 88 children born with a gestational age younger than 32 weeks between 2007 and 2009 who were screened and/or treated for ROP. The authors collected fetal cord blood (sample 1) and a blood sample at 4 to 6 weeks postnatal age (sample 2) for laboratory investigations. RESULTS: The non-ROP group had higher visfatin-1, visfatin-2, insulin-1, insulin-2, (IGF-I)-1, and (IGF-I)-2 levels than the ROP group (P < .001 for all levels). Change in visfatin was positively correlated with change in IGF-I and change in insulin (P < .001 and P < .001, respectively). Change in IGF-I was positively correlated with change in insulin (P < .001). CONCLUSION: Similar to IGF-I levels, determination of visfatin levels can be a predictive marker of ROP, but more studies are needed. PMID- 21634361 TI - Complete regression of retinoblastoma following intra-arterial chemotherapy. AB - A 22-month-old girl with Group C unilateral retinoblastoma demonstrated dramatic tumor regression after two infusions of 5 mg of intra-arterial melphalan as primary therapy. Complete tumor control without recurrence was noted at 1 year. Retinal and choroidal perfusion was intact and the electroretinogram improved following therapy. PMID- 21634362 TI - Role of the low-energy excited states in the radiolysis of aromatic liquids. AB - The contribution of the low-energy excited states to the overall product formation in the radiolysis of simple aromatic liquids--benzene, pyridine, toluene, and aniline--has been examined by comparison of product yields obtained in UV-photolysis and in gamma-radiolysis. In photolysis, these electronic excited states were selectively populated using UV-light excitation sources with various energies. Yields of molecular hydrogen and of "dimers" (biphenyl, bibenzyl, dipyridyl for benzene, toluene, pyridine, respectively, and of ammonia and diphenylamine for aniline) have been determined, since they are the most abundant radiolytic products. Negligibly small production of molecular hydrogen in the UV photolysis of aromatic liquids with excitation to energies of 4.88, 5.41, 5.79, and 6.70 eV and the lack of a scavenger effect suggest that this product originates from short-lived high-energy singlet states. A significant reduction in "dimer" radiation-chemical yields in the presence of scavengers such as anthracene or naphthalene indicates that the triplet excited states are important precursors to these products. The results for toluene and aniline suggest that efficient dissociation from the lowest-energy excited triplet state leads to noticeable "dimer" production. For benzene and pyridine, the lowest-energy triplet excited states are not likely to fragment into radicals because of the relatively large energy gap between the excited state level and corresponding bond dissociation energy. The "dimer" formation in the radiolysis of benzene and pyridine is likely to involve short-lived high-energy triplet states. PMID- 21634363 TI - Refinement of the alpha-U4O9 crystalline structure: new insight into the U4O9 -> U3O8 transformation. AB - The oxidation reaction of UO(2) into U(3)O(8) is studied as a function of the crystalline distortion of interstitial oxygen clusters, named cuboctahedra, which appear in U(4)O(9) and U(3)O(7) intermediate phases. For that purpose, the refinement of alpha-U(4)O(9) was performed because this phase undergoes a trigonal distortion from cubic beta-U(4)O(9) when the temperature is decreased. In alpha-U(4)O(9), the cuboctahedra can be described as crumpled sheets taken from a fragment of U(3)O(8). The manner by which the accumulation of crumpled sheets can lead to the formation of U(3)O(8) is discussed. PMID- 21634364 TI - Metallocorrole dendrimers: sensitive corrole-chromium(V)-nitride spin probes for studying the solution structure of dendrimers. AB - The corrole-chromium(V)-nitrido moiety is introduced as a uniquely sensitive EPR spin probe. We describe a series of corrole-centered poly(amidoamine) (PAMAM) dendrimers and the selective incorporation of the chromium(V)-nitrido moiety. The chromium-corrole cores are reactive toward both neutral and charged reagents, and the accessibility of the dendrimer cores enables easy manipulation of the spin probe. The spin probe reveals a pronounced solvent dependence of the solution phase structure of the dendrimers. PMID- 21634365 TI - Carbene stabilization of highly reactive main-group molecules. AB - This article highlights recent efforts of this laboratory in the stabilization of highly reactive, low-oxidation-state, main-group molecules using bulky N heterocyclic carbene ligands [L: = :C{N(2,6-Pr(i)(2)C(6)H(3))CH}(2); L': = :C{N(2,4,6-Me(3)C(6)H(2))CH}(2); L'': = :C{(i-Pr)NC(Me)}(2)]. The syntheses, structures, and computational studies of carbene-stabilized neutral diborenes [L:(H)B?B(H):L and L':(H)B?B(H):L'], a neutral Ga(6) octahedron (L'':Ga[Ga(4)Mes(4)]Ga:L''), disilicon (L:Si?Si:L), bis-silylene [L:(Cl)Si Si(Cl):L], dipnictogens (L:E-E:L, E = P, As; L':P-P:L'), and parent phosphinidene (L:PH) are discussed. Some of the unique challenges associated with this "carbene stabilization" strategy are also presented. PMID- 21634366 TI - Three-dimensional mapping of microenvironmental control of methyl rotational barriers. AB - Sterical (van der Waals-induced) rotational barriers of methyl groups are investigated theoretically, using ab initio and empirical force field calculations, for various three-dimensional microenvironmental conditions around the methyl group rotator of a model neopentane molecule. The destabilization (reducing methyl rotational barriers) or stabilization (increasing methyl rotational barriers) of the staggered conformation of the methyl rotator depends on a combination of microenvironmental contributions from (i) the number of atoms around the rotator, (ii) the distance between the rotator and the microenvironmental atoms, and (iii) the dihedral angle between the stator, rotator, and molecular environment around the rotator. These geometrical criteria combine their respective effects in a linearly additive fashion, with no apparent cooperative effects, and their combination in space around a rotator may increase, decrease, or leave the rotator's rotational barrier unmodified. This is exemplified in a geometrical analysis of the alanine dipeptide crystal where microenvironmental effects on methyl rotators' barrier of rotation fit the geometrical mapping described in the neopentane model. PMID- 21634367 TI - Competition between hydrogen bonding and dispersion interactions in the indole...pyridine dimer and (indole)2...pyridine trimer studied in a supersonic jet. AB - Structures of the indole...pyridine dimer and (indole)2...pyridine trimer have been investigated in a supersonic jet using resonant two-photon ionization (R2PI) and IR-UV double resonance spectroscopic techniques combined with quantum chemistry calculations. R2PI spectra of the dimer and the trimer recorded by electronic excitation of the indole moiety show that the red-shift in the band origin of the dimer with respect to the 0(0)(0) band of the monomer is larger compared to that of the trimer. The presence of only one conformer in the case of both the dimer and the trimer has been confirmed from IR-UV hole-burning spectroscopy. The structures of the dimer and the trimer have been determined from resonant ion dip infrared (RIDIR) spectra combined with ab initio as well as DFT/M05-2X and DFT/M06-2X calculations. It has been found that the dimer, observed in the experiment, has a V-shaped geometry stabilized by N-H...N and C H...N hydrogen bonding interactions, as well as C-H...pi and pi...pi dispersion interactions. The geometry of the trimer has been found to be a cyclic one stabilized by N-H...N, N-H...pi, C-H...pi, and C-H...N interactions. The most important finding of this current study is the observation of the mixed dimer and trimer, which are stabilized by hydrogen bonding as well as dispersion interactions. PMID- 21634368 TI - Synthesis of alpha-CN and alpha-CF3 N-heterocycles through tandem nucleophilic additions. AB - Using a readily available secondary aminoalkyne as starting material, a powerful strategy was discovered to prepare precursors of biologically important unnatural cyclic aminoacids and fluorinated N-heterocycles with important ring sizes (e.g., 5-7) in a one-pot reaction using two nucleophilic additions in a tandem fashion. PMID- 21634369 TI - Diastereoselective construction of 7-methylenebicyclo[3.2.1]oct-3-en-2-one derivatives by palladium-catalyzed cyclization of propargylic acetates with 2 oxocyclohex-3-enecarboxylates. AB - The reaction of propargylic acetates with 2-oxocyclohex-3-enecarboxylates in the presence of a palladium catalyst is described. Substituted 7 methylenebicyclo[3.2.1]oct-3-en-2-ones were synthesized in a highly diastereoselective manner. PMID- 21634371 TI - Vibrational spectroscopy and mass spectrometry for characterization of soft landed polyatomic molecules. AB - We report implementation of two powerful characterization tools, in situ secondary ion mass spectrometry (SIMS) and ex situ surface enhanced Raman spectroscopy (SERS), in analyzing surfaces modified by ion soft landing (SL). Cations derived from Rhodamine 6G are soft landed onto Raman-active silver colloidal substrates and detected using SERS. Alternatively and more conveniently, high-quality SERS data are obtained by spin coating a silver colloidal solution over the modified surface once SL is complete. Well-defined SERS features are observed for Rhodamine 6G in as little as 15 min of ion deposition. Deposition of ~3 pmo1 gave high-quality SERS spectra with characteristic spectroscopic responses being derived from just ~0.5 fmol of material. Confocal SERS imaging allowed the enhancement to be followed in different parts of deposited dried droplets on surfaces. Characteristic changes in Raman spectral features occur when Rhodamine 6G is deposited under conditions that favor gas-phase ion fragmentation. Simultaneous deposition of both the intact dye and its fragment ion occurs and is confirmed by SIMS analysis. The study was extended to other Raman active surfaces, including Au nanostar and Au coated Ni nanocarpet surfaces and to SL of other molecules including fluorescein and methyl red. Overall, the results suggest that combination of SERS and SIMS measurements are effective in the characterization of surfaces produced by ion SL with significantly enhanced molecular specificity. PMID- 21634372 TI - Fabrication and temperature-dependent field-emission properties of bundlelike VO2 nanostructures. AB - Bundlelike VO(2)(B) nanostructures were synthesized via a hydrothermal method, and VO(2)(M(1)/R) nanobundles were obtained after a heat-treatment process. Structural characterization shows that these nanobundles are self-assembled by VO(2) nanowires, and VO(2)(M(1)/R) nanobundles have better crystallinity. Temperature-dependent field-emission (FE) measurement indicates that FE properties of these two phases of nanobundles can both be improved by increasing the ambient temperature. Moreover, for the VO(2)(M(1)/R) nanobundles, their FE properties are also strongly dependent on the temperature-induced metal-insulator transitions process. Compared with poor FE properties found in the insulating phase, FE properties were significantly improved by increasing the temperature, and about a three-orders-of-magnitude increasing of the emission current density has been observed at a fixed field of 6 V/MUm. Work function measurement and density-functional theory calculations indicated that the decrease of work function with temperature is the main reason that caused the improvement of FE properties. These characteristics make VO(2)(M(1)/R) a candidate material for application of new type of temperature-controlled field emitters, whose emission density can be adjusted by ambient temperature. PMID- 21634373 TI - Limiting activity coefficients and gas-liquid partition coefficients of various solutes in piperidinium ionic liquids: measurements and LSER calculations. AB - This paper is a continuation of our systematic investigations on piperidinium ionic liquids and presents new data on activity coefficients at infinite dilution for 43 solutes: linear and branched alkanes, cycloalkanes, alkenes, alkynes, benzene, alkylbenzenes, alcohols, water, thiophene, tetrahyrdofuran (THF), methyl tert-butyl ether (MTBE), linear ethers, acetone, and linear ketones in the ionic liquid 1-butyl-1-methyl-piperidinium bis(trifluoromethylsulfonyl)imide, [BMPIP][NTf2]. The data were determined by gas-liquid chromatography (GLC) at temperatures from 308.15 to 358.15 K. These values were compared to those previously published for the bis-(trifluoromethylsulfonyl)imide-based ionic liquids. The partial molar excess enthalpies DeltaH1(E,infinity) and entropies DeltaS1(E,infinity) at infinite dilution were calculated from the experimental gamma13(infinity) values obtained over the temperature range. The values of the selectivities for different separation problems were calculated from gamma13(infinity) and compared to literature values for N-methyl-2-pyrrolidinone (NMP), sulfolane, and additional ionic liquids. Experimental limiting activity coefficients were used to calculate gas-IL partition coefficients of solutes, K(L). The modeling with specific linear solvation energy relationship (LSER) equations was performed for data obtained in this work and those reported earlier for 1-butyl-1-methylpiperidinium thiocyanate, [BMPIP][SCN]. PMID- 21634374 TI - Signaling kinetics of cyanobacterial phytochrome Cph1, a light regulated histidine kinase. AB - Cyanobacterial phytochrome 1 (Cph1) is a red/far-red light regulated histidine kinase, which together with its response regulator (Rcp1) forms a two-component light signaling system in Synechocystis 6803. In the present study we followed the in vitro autophosphorylation of Cph1 and the subsequent phosphotransfer to Rcp1 in different ionic milieus and following different light treatments. Both processes were red/far-red reversible with activity manifested in the Pr ground state (in darkness or after far-red irradiation) and with strongest activities being exhibited in the presence of Mn(2+). In vivo and in vitro assembled holoproteins in the Pr state displayed at least 4-fold higher efficiencies (k(cat)/K(m)) for autophosphorylation and phosphotransfer than the apoprotein or the holoprotein at photoequilibrium in red light. The reduced activities observed following red light treatments were consistent with the Pfr state being enzymatically inactive. Thus, both the rate of kinase autophosphorylation and the rate of phosphotransfer regulate the phosphorylation state of the response regulator, consistent with the rotary switch model regulating accessibility of the histidine target. PMID- 21634375 TI - Kinetic control and thermodynamic selection in the synthesis of atomically precise gold nanoclusters. AB - This work presents a combined approach of kinetic control and thermodynamic selection for the synthesis of monodisperse 19 gold atom nanoclusters protected by thiolate groups. The step of kinetic control allows the formation of a proper size distribution of initial size-mixed Au(n)(SR)(m) nanoclusters following the reduction of a gold precursor. Unlike the synthesis of Au(25)(SR)(18) nanoclusters, which involves rapid reduction of the gold precursor by NaBH(4) followed by size focusing, the synthesis of 19-atom nanoclusters requires slow reduction effected by a weaker reducing agent, borane-tert-butylamine complex. The initially formed mixture of nanoclusters then undergoes size convergence into a monodisperse product by means of a prolonged aging process. The nanocluster formula was determined to be Au(19)(SC(2)H(4)Ph)(13). This work demonstrates the importance of both kinetic control of the initial size distribution of nanoclusters prior to size focusing and subsequent thermodynamic selection of stable nanoclusters as the final product. PMID- 21634376 TI - Determination of cranberry phenolic metabolites in rats by liquid chromatography tandem mass spectrometry. AB - The glycosides of flavonoid, anthocyanins and A type proanthocyanidins in cranberry concentrate were characterized and quantified using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Cranberry concentrate (1 g/body weight) was orally gavaged to Fischer-344 rats (n = 6), and blood and urine samples were collected over 24 h periods. Quercetin, 3'-O-methylquercetin (isorhamnetin), myricetin, kaempferol, and proanthocyanidin dimer A2, together with thirteen conjugated metabolites of quercetin and methylquercetin and intact peonidin 3-O-galactoside and cyanidin 3-O-galactoside were identified in the rat urine after cranberry treatment. Very low levels of isorhamnetin (0.48 +/- 0.09 ng/mL) and proanthocyanidin dimer A2 (0.541 +/- 0.10 ng/mL) were found in plasma samples after 1 h of cranberry administration. Although no quercetin was detected in plasma, MRM analysis of the methanolic extract of urinary bladder showed that chronic administration of cranberry concentrate to rats resulted in accumulation of quercetin and isorhamnetin in the bladder. These results demonstrate that cranberry components undergo rapid metabolism and elimination into the urine of rats and are present in the urinary bladder tissue potentially allowing them to inhibit urinary bladder carcinogenesis. PMID- 21634377 TI - Identification of a potent, state-dependent inhibitor of Nav1.7 with oral efficacy in the formalin model of persistent pain. AB - Clinical human genetic studies have recently identified the tetrodotoxin (TTX) sensitive neuronal voltage gated sodium channel Nav1.7 (SCN9A) as a critical mediator of pain sensitization. Herein, we report structure-activity relationships for a novel series of 2,4-diaminotriazines that inhibit hNav1.7. Optimization efforts culminated in compound 52, which demonstrated pharmacokinetic properties appropriate for in vivo testing in rats. The binding site of compound 52 on Nav1.7 was determined to be distinct from that of local anesthetics. Compound 52 inhibited tetrodotoxin-sensitive sodium channels recorded from rat sensory neurons and exhibited modest selectivity against the hERG potassium channel and against cloned and native tetrodotoxin-resistant sodium channels. Upon oral administration to rats, compound 52 produced dose- and exposure-dependent efficacy in the formalin model of pain. PMID- 21634378 TI - 13C-labeled heparan sulfate analogue as a tool to study protein/heparan sulfate interactions by NMR spectroscopy: application to the CXCL12alpha chemokine. AB - Heparan sulfate (HS), a polysaccharide of the glycosaminoglycan family characterized by a unique level of complexity, has emerged as a key regulator of many fundamental biological processes. Although it has become clear that this class of molecules exert their functions by interacting with proteins, the exact modes of interaction still remain largely unknown. Here we report the engineering of a (13)C-labeled HS-like oligosaccharide with a defined oligosaccharidic sequence that was used to investigate the structural determinants involved in protein/HS recognition by multidimensional NMR spectroscopy. Using the chemokine CXCL12alpha as a model system, we obtained experimental NMR data on both the oligosaccharide and the chemokine that was used to obtain a structural model of a protein/HS complex. This new approach provides a foundation for further investigations of protein/HS interactions and should find wide application. PMID- 21634379 TI - Urea and guanidinium induced denaturation of a Trp-cage miniprotein. AB - Using a combination of experimental techniques (circular dichroism, differential scanning calorimetry, and NMR) and molecular dynamics simulations, we performed an extensive study of denaturation of the Trp-cage miniprotein by urea and guanidinium. The experiments, despite their different sensitivities to various aspects of the denaturation process, consistently point to simple, two-state unfolding process. Microsecond molecular dynamics simulations with a femtosecond time resolution allow us to unravel the detailed molecular mechanism of Trp-cage unfolding. The process starts with a destabilizing proline shift in the hydrophobic core of the miniprotein, followed by a gradual destruction of the hydrophobic loop and the alpha-helix. Despite differences in interactions of urea vs guanidinium with various peptide moieties, the overall destabilizing action of these two denaturants on Trp-cage is very similar. PMID- 21634380 TI - Introducing bioorthogonal functionalities into proteins in living cells. AB - Proteins are the workhorses of the cell, playing crucial roles in virtually every biological process. The revolutionary ability to visualize and monitor proteins in living systems, which is largely the result of the development of green fluorescence protein (GFP) and its derivatives, has dramatically expanded our understanding of protein dynamics and function. Still, GFPs are ill suited in many circumstances; one major drawback is their relatively large size, which can significantly perturb the functions of the native proteins to which they are fused. To bridge this gap, scientists working at the chemistry-biology interface have developed methods to install bioorthogonal functional groups into proteins in living cells. The bioorthogonal group is, by definition, a non-native and nonperturbing chemical group. But more importantly, the installed bioorthogonal handle is able to react with a probe bearing a complementary functionality in a highly selective fashion and with the cell operating in its physiological state. Although extensive efforts have been directed toward the development of bioorthogonal chemical reactions, introducing chemical functionalities into proteins in living systems remains an ongoing challenge. In this Account, we survey recent progress in this area, focusing on a genetic code expansion approach. In nature, a cell uses posttranslational modifications to append the necessary functional groups into proteins that are beyond those contained in the canonical 20 amino acids. Taking lessons from nature, scientists have chosen or engineered certain enzymes to modify target proteins with chemical handles. Alternatively, one can use the cell's translational machinery to genetically encode bioorthogonal functionalities, typically in the form of unnatural amino acids (UAAs), into proteins; this can be done in a residue-specific or a site specific manner. For studying protein dynamics and function in living cells, site specific modification by means of genetic code expansion is usually favored. A variety of UAAs bearing bioorthogonal groups as well as other functionalities have been genetically encoded into proteins of interest. Although this approach is well established in bacteria, tagging proteins in mammalian cells is challenging. A facile pyrrolysine-based system, which might potentially become the "one-stop shop" for protein modification in both prokaryotic and eukaryotic cells, has recently emerged. This technology can effectively introduce a series of bioorthogonal handles into proteins in mammalian cells for subsequent chemical conjugation with small-molecule probes. Moreover, the method may provide more precise protein labeling than GFP tagging. These advancements build the foundation for studying more complex cellular processes, such as the dynamics of important receptors on living mammalian cell surfaces. PMID- 21634381 TI - Preparation, crystal structures, and magnetic features for a series of dinuclear [Ni(II)Ln(III)] Schiff-base complexes: evidence for slow relaxation of the magnetization for the Dy(III) derivative. AB - A series of dinuclear [Ni(II)Ln(III)] Schiff-base complexes (using a Schiff-base dicompartmental ligand derived from o-vanillin [H(2)valpn = 1,3-propanediylbis(2 iminomethylene-6-methoxy-phenol)]) with Ln = La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, and a hydroxo-bridged tetranuclear [Ni(II)Yb(III)] are reported. The crystal structures have been solved for 10 dinuclear complexes revealing four arrangements for the dinuclear units, which are modulated by the coordinated solvent molecules and the nitrato-anion interactions. The magnetic behaviors have been investigated, and the nature of the Ni(II)-Ln(III) exchange interaction has been emphasized by comparison with the behavior of the related [Zn(II)Ln(III)] derivatives. This allowed for establishing that the interaction within these compounds is antiferromagnetic with the 4f ions of the beginning of the Ln series and turns ferromagnetic from Gd(III) toward the end of the series. AC susceptibility investigations clearly show the occurrence of slow relaxation processes of the magnetization close to 2 K for the dinuclear [Ni(II)Dy(III)] complex. PMID- 21634382 TI - Metal binding to ligand-containing peptide nucleic acids. AB - The substitution of nucleobases in nucleic acid duplexes with ligands that have high affinity for transition metal ions creates metal-binding sites at specific locations within the duplexes. Several studies on the incorporation of metal ions into DNA and peptide nucleic acid (PNA) duplexes have suggested that the stability constant of the metal complex formed within the duplexes is a primary determinant of the thermal stability of the duplexes. To understand this relationship, we have synthesized two PNA monomers that carry the same ligand, namely 8-hydroxyquinoline, but have this ligand attached differently to the PNA backbone. The PNA monomers have been incorporated into PNA duplexes. UV and CD spectroscopy and calorimetric studies of the 8-hydroxyquinoline-PNA duplexes showed that the effect of the stability of the metal complex on the PNA duplexes was significantly modulated by the steric relationship between the complex and the duplex. This information is useful for the construction of hybrid inorganic nucleic acid nanostructures. PMID- 21634383 TI - Study of the luminescent and magnetic properties of a series of heterodinuclear [Zn(II)Ln(III)] complexes. AB - Herein, we report the synthesis, structural investigation, and magnetic and photophysical properties of a series of 13 [Zn(II)Ln(III)] heterodinuclear complexes, which have been obtained employing a Schiff-base compartmental ligand derived from o-vanillin [H(2)valpn = 1,3-propanediylbis(2-iminomethylene-6 methoxy-phenol)]. The complexes have been synthesized starting from the [Zn(valpn)(H(2)O)] mononuclear compound and the corresponding lanthanide nitrates. The crystallographic investigation indicated two structural types: the first one, [Zn(H(2)O)(valpn)Ln(III)(O(2)NO)(3)], contains 10-coordinated Ln(III) ions, while in the second one, [Zn(ONO(2))(valpn)Ln(III)(H(2)O)(O(2)NO)(2)].2H(2)O, the rare earth ions are nine coordinated. The Zn(II) ions always display a square-pyramidal geometry. The first structural type encompasses the larger Ln ions (4f(0)-4f(9)), while the second is found for the smaller ions (4f(8)-4f(11)). The dysprosium derivative crystallizes in both forms. Luminescence studies for the heterodinuclear compounds containing Nd(III), Sm(III), Tb(III), Dy(III), and Yb(III) revealed that the [Zn(valpn)(H(2)O)] moiety acts as an antenna. The magnetic properties for the paramagnetic [Zn(II)Ln(III)] complexes have been investigated. PMID- 21634385 TI - Redox properties of Tanaka's water oxidation catalyst: redox noninnocent ligands dominate the electronic structure and reactivity. AB - [Ru(2)(OH)(2)(3,6-(t)Bu(2)Q)(2)(btpyan)](2+) ((t)Bu(2)Q, 3,6-di-tert-butyl-1,2 benzoquinone; btpyan, 1,8-bis(2,2':6',2''-terpyridyl)anthracene) is one of a handful of structurally well-defined homogeneous catalysts that can electrocatalytically oxidize water at room temperature. Unfortunately, the exact composition and the chemical properties of the redox intermediates leading to the catalytically competent species remains poorly resolved. On the basis of the UV vis spectra the catalyst was previously speculated to lose two protons spontaneously to form an intermediate containing the key O-O bond in water. We evaluated this mechanistic scenario computationally and found that the associated pK(a) values are in the range of 21, much too high to justify spontaneous deprotonation under experimental conditions of pH = 4. In later work, the O-O bond formation was speculated to occur after removal of two protons and two electrons. Extensive exploration of the various oxidation and protonation states that the diruthenium complex may access during catalyst activation reveals surprisingly complex electronic structure patterns in several redox intermediates: the quinone and tpy ligands become redox noninnocent, i.e., they participate actively in the electron transfer processes by temporarily storing redox equivalents. On the basis of this new insight into the electronic structure we propose a novel alternative explanation of the spectroscopic observations reported previously and characterize the electronic structure of the key intermediates in detail. Finally, the redox potential for the first two-electron oxidation is evaluated based on our proposed intermediates and predicted to be 0.411 V, which compares well with the experimentally observed broad two-electron wave at ~0.32 V. PMID- 21634384 TI - Construction of functionalized metallosupramolecular tetragonal prisms via multicomponent coordination-driven self-assembly. AB - A new approach for the construction of functionalized metallosupramolecular tetragonal prisms via multicomponent, coordination-driven, template-free self assembly is described. The combination of tetra-(4-pyridylphenyl)ethylene, a 90 degrees Pt(II) acceptor, and ditopic bipyridine or carboxylate ligands functionalized with hydroxyl or amine groups, hydrophobic alkyl chains, or electrochemically active ferrocene, yields a suite of seven self-assembled tetragonal prisms under mild conditions. These three-dimensional metallosupramolecules were characterized by multinuclear NMR ((31)P and (1)H) and mass spectrometry. Their shapes and sizes were established using Merck Molecular Force Field (MMFF) simulations. In addition, their approximate sizes were further supported by pulsed-field-gradient spin-echo (PGSE) NMR experiments. PMID- 21634386 TI - Theoretical study of oxidation of cyclohexane diol to adipic anhydride by [Ru(IV)(O)(tpa)(H2O)]2+ complex (tpa ? tris(2-pyridylmethyl)amine). AB - The catalytic conversion of 1,2-cyclohexanediol to adipic anhydride by Ru(IV)O(tpa) (tpa ? tris(2-pyridylmethyl)amine) is discussed using density functional theory calculations. The whole reaction is divided into three steps: (1) formation of alpha-hydroxy cyclohexanone by dehydrogenation of cyclohexanediol, (2) formation of 1,2-cyclohexanedione by dehydrogenation of alpha-hydroxy cyclohexanone, and (3) formation of adipic anhydride by oxygenation of cyclohexanedione. In each step the two-electron oxidation is performed by Ru(IV)O(tpa) active species, which is reduced to bis-aqua Ru(II)(tpa) complex. The Ru(II) complex is reactivated using Ce(IV) and water as an oxygen source. There are two different pathways of the first two steps of the conversion depending on whether the direct H-atom abstraction occurs on a C-H bond or on its adjacent oxygen O-H. In the first step, the C-H (O-H) bond dissociation occurs in TS1 (TS2-1) with an activation barrier of 21.4 (21.6) kcal/mol, which is followed by abstraction of another hydrogen with the spin transition in both pathways. The second process also bifurcates into two reaction pathways. TS3 (TS4-1) is leading to dissociation of the C-H (O-H) bond, and the activation barrier of TS3 (TS4-1) is 20.2 (20.7) kcal/mol. In the third step, oxo ligand attack on the carbonyl carbon and hydrogen migration from the water ligand occur via TS5 with an activation barrier of 17.4 kcal/mol leading to a stable tetrahedral intermediate in a triplet state. However, the slightly higher energy singlet state of this tetrahedral intermediate is unstable; therefore, a spin crossover spontaneously transforms the tetrahedral intermediate into a dione complex by a hydrogen rebound and a C-C bond cleavage. Kinetic isotope effects (k(H)/k(D)) for the electronic processes of the C-H bond dissociations calculated to be 4.9-7.4 at 300 K are in good agreement with experiment values of 2.8-9.0. PMID- 21634387 TI - Theoretical determination of the zero-field splitting in copper acetate monohydrate. AB - The zero-field splitting of the copper acetate monohydrate complex is studied using wave function based calculations. The anisotropy parameters extracted from highly correlated methods are in excellent agreement with the most accurate experimental results; in particular, the negative sign of the axial anisotropy parameter D is reproduced. During several decades, the interpretation of experimental data based on an analytical expression derived from perturbation theory led to a positive D-value. Although the validity of this expression is confirmed, it is explained that the incorrect attribution of a positive D is related to the assumption of an antiferromagnetic coupling between excited states. We have found in the present work that this coupling is actually ferromagnetic. The analysis of the various contributions to the anisotropy parameters shows that both spin-spin and spin-orbit couplings participate in the magnetic anisotropy of this complex. Although the anisotropy arising from the spin-spin coupling is essentially independent of the level of calculation, the zero-field-splitting parameters resulting from the spin-orbit coupling are strongly sensitive to the effects of dynamic correlation. This works provides important new insights into the physical origin of the zero-field-splitting parameters in copper dimers. PMID- 21634388 TI - Transferable SAFT-VR models for the calculation of the fluid phase equilibria in reactive mixtures of carbon dioxide, water, and n-alkylamines in the context of carbon capture. AB - The amine functional groups are fundamental building blocks of many molecules that are central to life, such as the amino acids, and to industrial processes, such as the alkanolamines, which are used extensively for gas absorption. The modeling of amines and of mixtures of amines with water (H(2)O) and carbon dioxide (CO(2)) is thus relevant to a number of applications. In this contribution, we use the statistical associating fluid theory for potentials of variable range (SAFT-VR) to describe the fluid phase behavior of ammonia + H(2)O + CO(2) and n-alkyl-1-amine + H(2)O + CO(2) mixtures. Models are developed for ammonia (NH(3)) and n-alkyl-1-amines up to n-hexyl-1-amine (CH(3)NH(2) to C(6)H(13)NH(2)). The amines are modeled as homonuclear chain molecules formed from spherical segments with additional association sites incorporated to mediate the effect of hydrogen-bonding interactions. The SAFT-VR approach provides a representation of the pure component fluid phase equilibria, on average, to within 1.48% of the experimental data in relative terms for the saturated liquid densities and vapor pressures. A simple empirical correlation is derived for the SAFT-VR parameters of the n -alkylamine series as a function of molecular weight. Aqueous mixtures of the amines are modeled using a model of water taken from previous work. The models developed for the mixtures are of high fidelity and can be used to calculate the binary fluid phase equilibrium of these systems to within 2.28% in relative terms for the temperature or pressure and 0.027 in absolute terms for the mole fraction. Regions of both vapor-liquid and liquid liquid equilibria are considered. We also consider the reactive mixtures of amines and CO(2) in aqueous solution. To model the reaction of CO(2) with the amine, an additional site is included on the otherwise nonassociating CO(2) model. The unlike interaction parameters for the NH(3) + H(2)O + CO(2) ternary mixture are obtained by comparison to the experimental data available for this system. The resulting model is found to correlate and predict the liquid-phase loading (moles of CO(2) per mole of amine) to within 0.091 of experimental data in absolute terms. The parameters describing the NH(3)-CO(2) interaction are then transferred to other n-alkyl-1-amines, and sample predictions of the fluid phase equilibria for the n-propyl-1-amine + H(2)O + CO(2), n-butyl-1-amine + H(2)O + CO(2), and n-hexyl-1-amine + H(2)O + CO(2) mixtures are presented. The latter mixture is found to exhibit regions of liquid-liquid immiscibility. PMID- 21634389 TI - Scaffolding, ladders, chains, and rare ferrimagnetism in intermetallic borides: synthesis, crystal chemistry and magnetism. AB - Single-phase polycrystalline samples and single crystals of the complex boride phases Ti(8)Fe(3)Ru(18)B(8) and Ti(7)Fe(4)Ru(18)B(8) have been synthesized by arc melting the elements. The phases were characterized by powder and single-crystal X-ray diffraction as well as energy-dispersive X-ray analysis. They are new substitutional variants of the Zn(11)Rh(18)B(8) structure type, space group P4/mbm (no. 127). The particularity of their crystal structure lies in the simultaneous presence of dumbbells which form ladders of magnetically active iron atoms along the [001] direction and two additional mixed iron/titanium chains occupying Wyckoff sites 4h and 2b. The ladder substructure is ca. 3.0 A from the two chains at the 4h, which creates the sequence chain-ladder-chain, establishing a new structural and magnetic motif, the scaffold. The other chain (at 2b) is separated by at least 6.5 A from this scaffold. According to magnetization measurements, Ti(8)Fe(3)Ru(18)B(8) and Ti(7)Fe(4)Ru(18)B(8) order ferrimagnetically below 210 and 220 K, respectively, with the latter having much higher magnetic moments than the former. However, the magnetic moment observed for Ti(8)Fe(3)Ru(18)B(8) is unexpectedly smaller than the recently reported Ti(9)Fe(2)Ru(18)B(8) ferromagnet. The variation of the magnetic moments observed in these new phases can be adequately understood by assuming a ferrimagnetic ordering involving the three different iron sites. Furthermore, the recorded hysteresis loops indicate a semihard magnetic behavior for the two phases. The highest H(c) value (28.6 kA/m), measured for Ti(7)Fe(4)Ru(18)B(8), lies just at the border of those of hard magnetic materials. PMID- 21634390 TI - Weak long-range correlated motions in a surface patch of ubiquitin involved in molecular recognition. AB - Long-range correlated motions in proteins are candidate mechanisms for processes that require information transfer across protein structures, such as allostery and signal transduction. However, the observation of backbone correlations between distant residues has remained elusive, and only local correlations have been revealed using residual dipolar couplings measured by NMR spectroscopy. In this work, we experimentally identified and characterized collective motions spanning four beta-strands separated by up to 15 A in ubiquitin. The observed correlations link molecular recognition sites and result from concerted conformational changes that are in part mediated by the hydrogen-bonding network. PMID- 21634391 TI - Redox-switching in a viologen-type adlayer: an electrochemical shell-isolated nanoparticle enhanced Raman spectroscopy study on Au(111)-(1*1) single crystal electrodes. AB - We reported the first application of in situ shell-isolated nanoparticle enhanced Raman spectroscopy (SHINERS) to an interfacial redox reaction under electrochemical conditions. We construct gap-mode sandwich structures composed of a thiol-terminated HS-6V6H viologen adlayer immobilized on a single crystal Au(111)-(1*1) electrode and covered by Au(60 nm)@SiO(2) core-shell nanoparticles acting as plasmonic antennas. We observed high-quality, potential-dependent Raman spectra of the three viologen species V(2+), V(+?), and V(0) on a well-defined Au(111) substrate surface and could map their potential-dependent evolution. Comparison with experiments on powder samples revealed an enhancement factor of the nonresonant Raman modes of ~3 * 10(5), and up to 9 * 10(7) for the resonance modes. The study illustrates the unique capability of SHINERS and its potential in the entire field of electrochemical surface science to explore structures and reaction pathways on well-defined substrate surfaces, such as single crystals, for molecular, (electro-)catalytic, bioelectrochemical systems up to fundamental double layer studies at electrified solid/liquid interfaces. PMID- 21634392 TI - Structural and electronic modulation of magnetic properties in a family of chiral iron coordination polymers. AB - The complexes FeL(2) [L = bidentate Schiff base ligands obtained from (R)-(+) alpha-phenylethanamine and 4-substituted salicylaldehydes, substituent R = H, (t)Bu, NO(2), OMe, CN, OH] react with ditopic proligands 1,4-pyrazine (pz) or 4,4'-bipyridine (bpy), to give a family of optically pure Fe(II) polymeric chain complexes of formula {FeL(2)(MU-pz)}(infinity) and {FeL(2)(MU-bpy)}(infinity). Crystallographic studies show that a range of structures are formed including unidirectional and bidirectional linear polymers and canted zigzag chains. Interchain interactions via pi-contacts and hydrogen bonding are also observed. SQuID magnetometry studies on all of the complexes reveal antiferromagnetic interactions, the magnitudes of which are rationalized on the basis of substituent electronic properties and bridging ligand identity. For complexes with bridging pz, the antiferromangnetic interaction is enhanced by electron releasing substituents on the Fe units, and this is accompanied by a contraction in the intrachain distance. For complexes bridged with the longer bpy the intrachain antiferromagnetic couplings are much weaker as a result of the longer intrachain distance. The magnetic data for this series of chain complexes follow a Bonner-Fisher 1D chain model, alongside a zero field splitting (ZFS) model for Fe(II) (S = 2) as appropriate. The intrachain antiferromagnetic coupling J values, g-factors, and the axial ZFS parameter D were obtained. PMID- 21634393 TI - Occurence of an octanuclear motif of uranyl isophthalate with cation-cation interactions through edge-sharing connection mode. AB - An uranyl isophthalate has been hydrothermally synthesized at 200 degrees C for 24 h, from a mixture of uranyl nitrate, isophthalic acid, and hydrazine in water. It was characterized by single-crystal analysis [triclinic, P 1, a = 7.3934(3) A, b = 13.3296(5) A, c = 15.4432(5) A, alpha = 111.865(2) degrees , beta = 90.637(2) degrees , gamma = 104.867(2) degrees , V = 1355.49(9) A(3)] and different spectroscopic techniques (Raman, IR-ATR, UV-visible). The 3D structure of the phase (UO(2))(8)O(2)(OH)(4)(H(2)O)(4)(1,3-bdc)(4).4H(2)O (1,3-bdc = 1,3 benzenedicarboxylate) reveals octanuclear units based on the association of 7 fold coordinated uranyl cations (pentagonal bipyramid) involving a rare case of cation-cation interaction together with edge-sharing polyhedral connection mode. UV-visible absorption spectroscopy confirmed that uranium was only involved in the structure as uranyl forms (excluding the presence of tetravalent or pentavalent uranium). Additionally, MU-Raman and IR-ATR experiments allowed assigning four uranyl contributions to the four types of uranyl entities in the structure, in agreement with the XRD analysis. PMID- 21634394 TI - Room-temperature ionic liquids discerned via nitroxyl spin probe dynamics. AB - The temperature dependence of the rotational correlation times, tau(c), of the nitroxide spin probes TEMPO, TEMPOL, TEMPAMINE, and Fremy's salt in the ionic liquids 1-ethyl-3-methylimidazolium ethylsulfate, 1-butyl-3-methylimidazolium hexafluorophosphate, 1-ethyl-3-methylimidazolium tetrafluoroborate, and 1-butyl-3 methylimidazolium tetrafluoroborate is scrutinized. The rotation correlation times vary between 54 and 1470 ps at 300 K. Within a temperature range of 280-380 K, the rotational tumbling is well described by the extended Debye-Stokes Einstein law. The hydrodynamic radii are smaller than the geometrical radii though. This discrepancy can partly be accounted for by microviscosity effects and deviations from the spherical shape. This study is distinguished from similar studies by the fact that proton superhyperfine coupling constants could be resolved for all nitroxides in the ionic liquids by carefully optimizing the experimental protocol. As a consequence, many rotational correlation times reported here are smaller than those found previously. Furthermore, the temperature dependence of the nitrogen ESR coupling constants is reported and discussed in detail. A surprising effect of adventitious water is reported for TEMPAMINE. PMID- 21634395 TI - Highly regioselective [3 + 2] annulation of azomethine imines with 1-alkynyl Fischer carbene complexes to functionalized N,N-bicyclic pyrazolidin-3-ones. AB - The highly regioselective [3 + 2] cycloaddition of azomethine imines to 1-alkynyl Fischer carbene complexes has been successfully realized under mild conditions. Oxidative demetalation of the newly formed pyrazolo-pyrazolone carbene complexes with pyridine-N-oxide or ceric ammonium nitrate efficiently afforded pyrazolo pyrazolone derivatives as well as cycloprop-2-enone and trisubstituted 1H pyrazoles in some cases, providing a novel route to versatile functionalized N,N bicyclic pyrazolidin-3-ones. PMID- 21634396 TI - Discovery and characterization of 6-{4-[3-(R)-2-methylpyrrolidin-1 yl)propoxy]phenyl}-2H-pyridazin-3-one (CEP-26401, irdabisant): a potent, selective histamine H3 receptor inverse agonist. AB - Optimization of a novel series of pyridazin-3-one histamine H(3) receptor (H(3)R) antagonists/inverse agonists identified 6-{4-[3-(R)-2-methylpyrrolidin-1 yl)propoxy]phenyl}-2H-pyridazin-3-one (8a, CEP-26401; irdabisant) as a lead candidate for potential use in the treatment of attentional and cognitive disorders. 8a had high affinity for both human (K(i) = 2.0 nM) and rat (K(i) = 7.2 nM) H(3)Rs with greater than 1000-fold selectivity over the hH(1)R, hH(2)R, and hH(4)R histamine receptor subtypes and against an in vitro panel of 418 G protein-coupled receptors, ion channels, transporters, and enzymes. 8a demonstrated ideal pharmaceutical properties for a CNS drug in regard to water solubility, permeability and lipophilicity and had low binding to human plasma proteins. It weakly inhibited recombinant cytochrome P450 isoforms and human ether-a-go-go-related gene. 8a metabolism was minimal in rat, mouse, dog, and human liver microsomes, and it had good interspecies pharmacokinetic properties. 8a dose-dependently inhibited H(3)R agonist-induced dipsogenia in the rat (ED(50) = 0.06 mg/kg po). On the basis of its pharmacological, pharmaceutical, and safety profiles, 8a was selected for preclinical development. The clinical portions of the single and multiple ascending dose studies assessing safety and pharmacokinetics have been completed allowing for the initiation of a phase IIa for proof of concept. PMID- 21634397 TI - Topology and accessibility of the transmembrane helices and the sensory site in the bifunctional transporter DcuB of Escherichia coli. AB - C(4)-Dicarboxylate uptake transporter B (DcuB) of Escherichia coli is a bifunctional transporter that catalyzes fumarate/succinate antiport and serves as a cosensor of the sensor kinase DcuS. Sites and domains of DcuB were analyzed for their topology relative to the cytoplasmic or periplasmic side of the membrane and their accessibility to the water space. For the topology studies, DcuB was fused at 33 sites to the reporter enzymes PhoA and LacZ that are only active when located in the periplasm or the cytoplasm, respectively. The ratios of the PhoA and LacZ activities suggested the presence of 10 or 11 hydrophilic loops, and 11 or 12 alpha-helical transmembrane domains (TMDs). The central part of DcuB allowed no clear topology prediction with LacZ/PhoA fusions. The sites of DcuB accessible to the hydrophilic thiol reagent 4-acetamido-4'-maleimidylstilbene 2,2'-disulfonate (AMS) were determined with variants of DcuB that carried single Cys residues. After intact cells were labeled with the membrane-impermeable AMS, denatured cells were differentially labeled with the thiol reagent polyethylene glycol-maleimide (PEGmal) and analyzed for a mass shift. From 35 positions 17 were accessible to AMS in intact bacteria. The model derived from topology and accessibility suggests 12 TMDs for DcuB and a waterfilled cavity in its central part. The cavity ends with a cytoplasmic lid accessible to AMS from the periplasmic side. The sensory domain of DcuB is composed of cytoplasmic loop XI/XII and a membrane integral region with the regulatory residues Thr396/Asp398 and Lys353. PMID- 21634398 TI - Expanding the one-dimensional CdS-CdSe composition landscape: axially anisotropic CdS 1-x Se x nanorods. AB - We report the synthesis and characterization of CdS(1-x)Se(x) nanorods with axial anisotropy. These nanorods were synthesized via single injection of a mixture of trioctylphosphine sulfur and selenium precursors to a cadmium-phosphonate complex at high temperature. Transmission electron microscopy shows nanoparticle morphology changes with relative sulfur and selenium loading. When the synthetic selenium loading is between 5% and 10% of total chalcogenides, the nanorods exhibit pronounced axial anisotropy characterized by a thick "head" and a thin "tail". The nanorods' band gap red shifts with increasing selenium loading. X-ray diffraction reveals that CdS(1-x)Se(x) nanorods have a wurtzite crystal structure with a certain degree of alloying. High-resolution and energy-filtered transmission electron microscopy and energy-dispersive X-ray spectroscopy confirm the head of the anisotropic nanorods is rich in selenium, whereas the tail is rich in sulfur. Time evolution and mechanistic studies confirm the nanorods form by quick growth of the CdSe-rich head, followed by slow growth of the CdS-rich tail. Metal photodeposition reactions with 575 nm irradiation, which is mostly absorbed by the CdSe-rich segment, show effective electronic communication between the nanorod head and tail segments. PMID- 21634399 TI - Diffusion-limited energy transfer in blends of oligofluorenes with an anthracene derivative. AB - Organic semiconductor devices such as light-emitting diodes and solar cells frequently comprise a blend of molecular or polymeric materials. Consequently, resonant energy transfer between the components plays a major role in determining device performance. Energy transfer may take place through either single-step donor-acceptor transfer, realized for example as Forster transfer, or as a sequence of donor-donor transfers toward the acceptor site. Here we use a well defined model system comprising an oligofluorene trimer, pentamer, or heptamer as the donor in combination with an anthracene derivative as the acceptor in order to study the rate and mechanism of energy transfer in thin films by time-resolved photoluminescence spectroscopy. We find the transfer process to be entirely dominated by sequential donor-donor transfer. In addition, we observe a strong dependence on oligomer length with an optimum energy transfer rate for the pentamer. PMID- 21634400 TI - Direct observation of a dark state in lycopene using pump-DFWM. AB - We apply pump-degenerate four-wave-mixing (pump-DFWM) for the investigation of the ultrafast internal relaxation of the excited states of lycopene. A unique feature in the pump-DFWM signal, appearing at small temporal delays between the initial pump pulse and the DFWM sequence, provides direct evidence for the participation of an additional excited state located between the S(2) and S(1) states. Our experimental findings are corroborated by a detailed numerical simulation of lycopene's pump-DFWM signal using the Brownian oscillator model. A very fast dynamics directly after excitation of the S(2) state manifests as a component populated with a time constant of about 20 fs and which decays to S(1) with a lifetime of 110 fs. This ultrafast dynamics is discussed under the light of several different models suggested for the relaxation pathway of carotenoids. In this context, we show that the dynamics can be explained in terms of a dark electronic state between the S(2) and S(1) states. PMID- 21634401 TI - Experimental investigations of a partial Ru-O bond during the metal-ligand bifunctional addition in Noyori-type enantioselective ketone hydrogenation. AB - The transition state for the metal-ligand bifunctional addition step in Noyori's enantioselective ketone hydrogenation was investigated using intramolecular trapping experiments. The bifunctional addition between the Ru dihydride trans [Ru((R)-BINAP)(H)(2)((R,R)-dpen)] and the hydroxy ketone 4 HOCH(2)C(6)H(4)(CO)CH(3) at -80 degrees C exclusively formed the corresponding secondary ruthenium alkoxide trans-[Ru((R)-BINAP)(H)(4 HOCH(2)C(6)H(4)CH(CH(3))O)((R,R)-dpen)]. Combined with the results of control experiments, this observation provides strong evidence for the formation of a partial Ru-O bond in the transition state. PMID- 21634402 TI - Structural studies of the monolayers and bilayers formed by a novel cholesterol phospholipid chimera. AB - Langmuir isotherm, neutron reflectivity, and small angle neutron scattering studies have been conducted to characterize the monolayers and vesicular bilayers formed by a novel chimeric phospholipid, ChemPPC, that incorporates a cholesteryl moeity and a C-16 aliphatic chain, each covalently linked via a glycerol backbone to phosphatidylcholine. The structures of the ChemPPC monolayers and bilayers are compared against those formed from pure dipalmitoylphoshatidylcholine (DPPC) and those formed from a 60:40 mol % mixture of DPPC and cholesterol. In accord with previous findings showing that very similar macroscopic properties were exhibited by ChemPPC and 60:40 mol % DPPC/cholesterol vesicles, it is found here that the chimeric lipid and lipid/sterol mixture have very similar monolayer structures (each having a monolayer thickness of ~26 A), and they also form vesicles with similar lamellar structure, each having a bilayer thickness of ~50 A and exhibiting a repeat spacing of ~65 A. The interfacial area of ChemPPC, however, is around 10 A(2) greater than that of the combined DPPC/cholesterol unit in the mixed lipid monolayer (viz., 57 +/- 1 vs 46 +/- 1 A(2), at 35 mN.m(-1)), and this difference in area is attributed to the succinyl linkage which joins the ChemPPC steroid and glyceryl moieties. The larger area of the ChemPPC is reflected in a slightly thicker monolayer solvent distribution width (9.5 vs 9 A for the DPPC/cholesterol system) and by a marginal increase in the level of lipid headgroup hydration (16 vs 13 H(2)O per lipid, at 35 mN.m(-1)). PMID- 21634403 TI - The conversion of a phenol to an aniline occurs in the biochemical formation of the 1-(4-aminophenyl)-1-deoxy-D-ribitol moiety in methanopterin. AB - Recent work has demonstrated that 4-hydroxybenzoic acid is the in vivo precursor to the 1-(4-aminophenyl)-1-deoxy-D-ribitol (APDR) moiety present in the C(1) carrier coenzyme methanopterin present in the methanogenic archaea. For this transformation to occur, the hydroxyl group of the 4-hydroxybenzoic acid must be replaced with an amino group at some point in the biosynthetic pathway. Using stable isotopically labeled precursors and liquid chromatography with electrospray-ionization mass spectroscopy, the first step of this transformation in Methanocaldococcus jannaschii occurs by the reaction of 4-hydroxybenzoic acid with phosphoribosyl pyrophosphate (PRPP) to form 4-(beta-d ribofuranosyl)hydroxybenzene 5'-phosphate (beta-RAH-P). The beta-RAH-P then condenses with l-aspartate in the presence of ATP to form 4-(beta-d ribofuranosyl)-N-succinylaminobenzene 5'-phosphate (beta-RFSA-P). Elimination of fumarate from beta-RFSA-P produces 4-(beta-D-ribofuranosyl)aminobenzene 5' phosphate (beta-RFA-P), the known precursor to the APDR moiety of methanopterin [White, R. H. (1996) Biochemistry 35, 3447-3456]. This work represents the first biochemical example of the conversion of a phenol to an aniline. PMID- 21634404 TI - pH-sensitive C-ON bond homolysis of alkoxyamines of imidazoline series with multiple ionizable groups as an approach for control of nitroxide mediated polymerization. AB - Recently, a new concept of pH-switchable agents for reversible addition fragmentation chain transfer (RAFT) polymerization has been introduced by Benaglia et al. (J. Am. Chem. Soc.2009, 131, 6914-6915). In this paper we extended the concept of pH-switchable mediators to nitroxide mediated polymerization (NMP) by employing nitroxides with basic or acidic groups as controlling agents. Four alkoxyamines, the derivatives of 2-(4-(dimethylamino)-2 ethyl-5,5-dimethyl-2-(pyridin-4-yl)-2,5-dihydro-1H-imidazol-1-oxyl and 2-(2 carboxyethyl)-5,5-diethyl-2,4-dimethyl-2,5-dihydro-1H-imidazol-1-oxyl, have been prepared. The influence of pH on alkoxyamine homolysis rate constants (k(d)) and on the nitroxide-alkyl radical recombination rate constants (k(c)) was studied. All alkoxyamines under study as well as the parent nitroxides have several basic groups, which under pH variation can undergo consecutive protonation. It was shown that the k(d) value under basic conditions are significantly (up to 15 fold) higher than in acidic solution at the same temperature, whereas the k(c) value in basic solutions decrease by a factor of 2 only. The efficiency of NMP is known to be dependent on k(d) and k(c), both constants being dependent on the monomer structure; therefore the performance of NMP of different monomers in the controlled mode requires different conditions. It is shown that the pH value crucially affects the polymerization regime, changing it from the controlled to the uncontrolled mode. The controlled regime of NMP of different hydrophilic monomers (sodium 4-styrenesulphonate and acrylamide) in aqueous solution under mild conditions (90 degrees C) can be achieved using the same alkoxyamine by the variation of the pH value. The chain length of polymers depends on pH value during the polymerization. PMID- 21634405 TI - Raman chemical imaging of chromate reduction sites in a single bacterium using intracellularly grown gold nanoislands. AB - Imaging live molecular events within micro-organisms at single-cell resolution would deliver valuable mechanistic information much needed in understanding key biological processes. We present a surface-enhanced Raman (SERS) chemical imaging strategy as a first step toward exploring the intracellular bioreduction pockets of toxic chromate in Shewanella. In order to achieve this, we take advantage of an innate reductive mechanism in bacteria of reducing gold ions into intracellular gold nanoislands, which provide the necessary enhancement for SERS imaging. We show that SERS has the sensitivity and selectivity not only to identify but also to differentiate between the two stable valence forms of chromate in cells. The imaging platform was used to understand intracellular metal reduction activities in a ubiquitous metal-reducing organism, Shewanella oneidensis MR-1, by mapping chromate reduction. PMID- 21634406 TI - A simulation study on nanoscale holes generated by gold nanoparticles on negative lipid bilayers. AB - Understanding the interactions of gold nanoparticles (AuNPs) with cellular compartments, especially cell membranes, is of fundamental importance in obtaining their control in biomedical applications. An effort is made in this paper to investigate the interactions of 2.2 nm core AuNPs with negative model bilayer membranes by coarse-grained (CG) molecular dynamics (MD) simulation. The CG model of lipid bilayer was taken from Marrink et al. ( J. Phys. Chem. B 2004, 108, 750-760 ), whereas the CG AuNPs model was developed on the basis of both atomistic MD simulations and experimental data. It was found that AuNPs functionalized with cationic ligands penetrated into the negative bilayer membranes and generated significant disruptions on bilayers. The lipids surrounding the nanoparticle were highly disordered and the bulk surface of the bilayer exhibits some defective areas. Most importantly, it is observed that a nanoscale hole can be formed and expanded spontaneously on the peripheral regions of the 20 * 20 nm bilayer. The expansion of the hole is on the time scale of hundreds of nanosceonds. The fully expanded hole had a radius of ~5.5 nm and could transport water molecules at a rate of up to ~1100 molecule/ns. However holes could not be formed on a larger bilayer (28 * 28 nm). The factors that can eliminate hole formation on the bilayer also include the decrease of cationic lignads on the AuNP, the reduction of negative lipids in the bilayer, the release of bilayer surface tension, the lowering of temperature, and the addition of a high concentration of salt. The results suggest that a hole can only be formed on living cell membranes under extreme conditions. PMID- 21634407 TI - The shape effect of mesoporous silica nanoparticles on biodistribution, clearance, and biocompatibility in vivo. AB - In our previous study we reported that the interaction of nanoparticles with cells can be influenced by particle shape, but until now the effect of particle shape on in vivo behavior remained poorly understood. In the present study, we control the fabrication of fluorescent mesoporous silica nanoparticles (MSNs) by varying the concentration of reaction reagents especially to design a series of shapes. Two different shaped fluorescent MSNs (aspect ratios, 1.5, 5) were specially designed, and the effects of particle shape on biodistribution, clearance and biocompatibility in vivo were investigated. Organ distributions show that intravenously administrated MSNs are mainly present in the liver, spleen and lung (>80%) and there is obvious particle shape effects on in vivo behaviors. Short-rod MSNs are easily trapped in the liver, while long-rod MSNs distribute in the spleen. MSNs with both aspect ratios have a higher content in the lung after PEG modification. We also found MSNs are mainly excreted by urine and feces, and the clearance rate of MSNs is primarily dependent on the particle shape, where short-rod MSNs have a more rapid clearance rate than long-rod MSNs in both excretion routes. Hematology, serum biochemistry, and histopathology results indicate that MSNs would not cause significant toxicity in vivo, but there is potential induction of biliary excretion and glomerular filtration dysfunction. These findings may provide useful information for the design of nanoscale delivery systems and the environmental fate of nanoparticles. PMID- 21634408 TI - Nanoprobes: quantitatively detecting the femtogram level of arsenite ions in live cells. AB - In this report, nanoprobes which could detect the femtogram level of arsenite ions in subcellular organelle of live cells are disclosed. The nanoprobes are composed of ssDNA and single-wall carbon nanotubes (SWCNTs), and the ssDNA is marked by a dye molecule. In a live cell, trace arsenite ions could interact with nanoprobes and significantly decrease the emission of the nanoprobes. With the help of a confocal microscope and cryo-electron microscopy, the lysosome target of the arsenite ion and nanoprobe is well described in high spatial resolution. PMID- 21634409 TI - Ordered polymeric microhole array made by selective wetting and applications for electrochemical microelectrode array. AB - In this paper, we report the microelectrode array fabrication using selective wetting/dewetting of polymers on a chemical pattern which is a simple and convenient method capable of creating negative polymeric replicas using polyethylene glycol (PEG) as a clean and nontoxic sacrificial layer. The fabricated hole-patterned polypropylene film on gold demonstrated enhanced electrochemical properties. The chemical pattern is fabricated by microcontact printing using octadecanethiol (ODT) as an ink on gold substrate. When PEG is spin-cast on the chemical pattern, PEG solution selectively dewets the ODT patterned areas and wets the remaining bare gold areas, leading to the formation of arrayed PEG dots. A negative replicas of the PEG dot array is obtained by spin coating of polypropylene (PP) solution in hexane which preferentially interacts with the hydrophobic ODT region on the patterned gold surface. The arrayed PEG dots are not affected the during PP spin-coating step because of their intrinsic immiscibility. Consequently, the hole-patterned PP film is obtained after PEG removal. The electrochemical signal of the PP film demonstrates the negligible leakage current by high dielectric and self-healing of defects on the chemical pattern by the polymer. This method is applicable to fabrication of microelectrode arrays and possibly can be employed to fabricate a variety of functional polymeric structures, such as photomasks, arrays of biomolecules, cell arrays, and arrays of nanomaterials. PMID- 21634410 TI - Viscosity of liquid perfluoroalkanes and perfluoroalkylalkane surfactants. AB - As part of a systematic study of the thermophysical properties of two important classes of fluorinated organic compounds (perfluoroalkanes and perfluoroalkylalkanes), viscosity measurements of four n-perfluoroalkanes and five perfluoroalkylalkanes have been carried out at atmospheric pressure and over a wide range of temperatures (278-353 K). From the experimental results the contribution to the viscosity from the CF(2) and CF(3) groups as a function of temperature have been estimated. Similarly, the contributions for CH(2) and CH(3) groups in n-alkanes have been determined using literature data. For perfluoroalkylalkanes, the viscosity results were interpreted in terms of the contributions of the constituent CF(2), CF(3), CH(2), and CH(3) groups, the deviations from ideality on mixing hydrogenated and fluorinated chains, and the contribution due to the formation of the CF(2)-CH(2) bond. A standard empirical group contribution method (Sastri-Rao method) has also been used to estimate the viscosities of the perfluoroalkylalkanes. Finally, to obtain molecular level insight into the behavior of these molecules, all-atom molecular dynamics simulations have been performed and used to calculate the densities and viscosities of the perfluoroalkylalkanes studied. Although both quantities are underestimated compared to the experimental data, with the viscosities showing the largest deviations, the trends observed in the experimental viscosities are captured. PMID- 21634411 TI - Extreme hardening of PDMS thin films due to high compressive strain and confined thickness. AB - Polymers confined to small dimensions and that undergo high strains can show remarkable nonlinear mechanics, which must be understood to accurately predict the functioning of nanoscale polymer devices. In this paper we describe the determination of the mechanical properties of ultrathin polydimethylsiloxane (PDMS) films undergoing large strains, using atomic force microscope (AFM) indentation with a spherical tip. The PDMS was molded into extremely thin films of variable thickness and adhered to a hard substrate. We found that for films below 1 MUm in thickness the Young's modulus increased with decreasing sample thickness with a power law exponent of 1.35. Furthermore, as the soft PDMS film was indented, significant strain hardening was observed as the indentation depth approached 45% of the sample thickness. To properly quantify the nonlinear mechanical measurements, we utilized a pointwise Hertzian model which assumes only piecewise linearity on the part of the probed material. This analysis revealed three regions within the material. A linear region with a constant Young's modulus was seen for compression up to 45% strain. At strains higher than 45%, a marked increase in Young's modulus was measured. The onset of strain induced stiffening is well modeled by finite element modeling and occurs as stress contours expanding from the probe and the substrate overlap. A third region of mechanical variation occurred at small indentations of less than 10 nm. The pointwise Young's modulus at small indentations was several orders of magnitude higher than that in the linear elasticity region; we studied and ruled out causes responsible for this phenomenon. In total, these effects can cause thin elastomer films to become extremely stiff such that the measured Young's modulus is over a 100-fold higher than the bulk PDMS. Therefore, the mechanics of a polymer can be changed by adjusting the geometry of a material, in addition to changing the material itself. In addition to understanding the mechanics of thin polymer films, this work provides an excellent test of experimental techniques to measure the mechanics of other nonlinear and heterogeneous materials such as biological cells. PMID- 21634412 TI - Detection of pure chemical vapors in a thermally cycled porous silica photonic crystal. AB - The condensation and evaporation of vapors of isopropanol, heptane, and cyclohexane in mesoporous silica photonic crystals are monitored by optical reflection spectroscopy as a function of sensor temperature. The spectral position of the stop band shifts to the red upon analyte adsorption, and it shifts to the blue as the sensor is heated and analyte evaporates from the porous nanostructure. The hysteresis of the optical response as the temperature of the sensor is cycled between 25 and 80 degrees C is characteristic of each analyte for partial pressures between 0 and 7.5 Torr. These characteristic hysteresis loops allow identification of the three analytes. The temporal response of the sensor is studied as a function of heating rate and analyte concentration in a flowing stream of analyte vapor, and it is compared with the equilibrium adsorption isotherms of the sensor. The ability of the temporal data to identify the analytes is attributed to differences in diffusion and adsorption properties of each analyte within the mesoporous silica sensor. PMID- 21634413 TI - Cytochrome c/polyelectrolyte multilayers investigated by E-QCM-D: effect of temperature on the assembly structure. AB - Protein multilayers, consisting of cytochrome c (cyt c) and poly(aniline sulfonic acid) (PASA), are investigated by electrochemical quartz crystal microbalance with dissipation monitoring (E-QCM-D). This technique reveals that a four-bilayer assembly has rather rigid properties. A thickness of 16.3 +/- 0.8 nm is calculated with the Sauerbrey equation and is found to be in good agreement with a viscoelastic model. The electroactive amount of cyt c is estimated by the deposited mass under the assumption of 50% coupled water. Temperature-induced stabilization of the multilayer assembly has been investigated in the temperature range between 30 and 45 degrees C. The treatment results in a loss of material and a contraction of the film. The electroactive amount of cyt c also decreases during heating and remains constant after the cooling period. The contraction of the film is accompanied by the enhanced stability of the assembly. In addition, it is found that cyt c and PASA can be assembled at higher temperatures, resulting in the formation of multilayer systems with less dissipation. PMID- 21634414 TI - Antiplasmodial sesquiterpene alkaloids from the roots of Maytenus mekongensis. AB - Eight new sesquiterpene alkaloids (1-8) and four known sesquiterpene alkaloids (9 12) have been isolated from the roots of Maytenus mekongensis. Structures were determined using extensive spectroscopic methods. The relative configuration of 7 epi-mekongensine (2) was established by single-crystal X-ray crystallographic analysis. The alkaloids were evaluated for antiplasmodial activity against Plasmodium falciparum, K1 strain, and for cytotoxicity using a panel of cell lines. PMID- 21634415 TI - Bioactive compounds from the aerial parts of Brachystemma calycinum and structural revision of an octacyclopeptide. AB - Four new cyclic peptides, brachystemins F-I (1-4), and 11 known compounds were isolated from the aerial parts of Brachystemma calycinum. The absolute configurations of compounds 1-4 were assigned using Marfey's method. The structure of compound 5 was revised from cyclo(Pro1-Phe2-Leu3-Ala4-Thr5-Pro6-Ala7 Gly8) to cyclo(Pro1-Pro2-Ala3-Gly4-Leu5-Ala6-Thr7-Phe8) with QTOF/MS and X-ray diffraction analysis. The N-containing compounds were assessed for their inhibitory effects on the secretion of monocyte chemokine ligand 2 (CCL-2), interleukin 6 (IL-6), and collagen IV against high-glucose-stimulated mesangial cells. Compound 5 was evaluated for its effects on collagen I, reactive oxygen species (ROS), superoxide anion (O2(*-)) production, and cell viability in mesangial cells, and on nitric oxide (NO) production in macrophage cells. PMID- 21634416 TI - Spectroscopic characterization of mononitrosyl complexes in heme--nonheme diiron centers within the myoglobin scaffold (Fe(B)Mbs): relevance to denitrifying NO reductase. AB - Denitrifying NO reductases are evolutionarily related to the superfamily of heme- copper terminal oxidases. These transmembrane protein complexes utilize a heme nonheme diiron center to reduce two NO molecules to N(2)O. To understand this reaction, the diiron site has been modeled using sperm whale myoglobin as a scaffold and mutating distal residues Leu-29 and Phe-43 to histidines and Val-68 to a glutamic acid to create a nonheme Fe(B) site. The impact of incorporation of metal ions at this engineered site on the reaction of the ferrous heme with one NO was examined by UV-vis absorption, EPR, resonance Raman, and FTIR spectroscopies. UV--vis absorption and resonance Raman spectra demonstrate that the first NO molecule binds to the ferrous heme, but while the apoproteins and Cu(I)- or Zn(II)-loaded proteins show characteristic EPR signatures of S = 1/2 six-coordinate heme {FeNO}(7) species that can be observed at liquid nitrogen temperature, the Fe(II)-loaded proteins are EPR silent at >=30 K. Vibrational modes from the heme [Fe-N-O] unit are identified in the RR and FTIR spectra using (15)NO and (15)N(18)O. The apo and Cu(I)-bound proteins exhibit nu(FeNO) and nu(NO) that are only marginally distinct from those reported for native myoglobin. However, binding of Fe(II) at the Fe(B) site shifts the heme nu(FeNO) by 17 cm(-1) and the nu(NO) by -50 cm(-1) to 1549 cm(-1). This low nu(NO) is without precedent for a six-coordinate heme {FeNO}(7) species and suggests that the NO group adopts a strong nitroxyl character stabilized by electrostatic interaction with the nearby nonheme Fe(II). Detection of a similarly low nu(NO) in the Zn(II)-loaded protein supports this interpretation. PMID- 21634417 TI - Structures of neutral and protonated water clusters confined in predesigned hosts: a quantum mechanical/molecular mechanical study. AB - Hybrid quantum mechanical/molecular mechanical (QM/MM) calculations have been carried out to investigate the structures of the neutral (H(2)O)(21) and protonated H(+)(H(2)O)(21) clusters confined in the crystal hosts. The influence of other cocrystallized species and the local electrostatic environments of the crystal hosts on the structures of water clusters has been analyzed. For the neutral (H(2)O)(21) cluster in the tetrahedral host, its low-lying structures are found to exist as a dodecahedral cage with one interior water molecule, which is in good accord with the corresponding X-ray data. The confined (H(2)O)(21) cluster possesses the main structural features of the lowest-energy structure of the free (H(2)O)(21) cluster in the gas phase. For the protonated H(+)(H(2)O)(21) cluster confined in the cubic cavity, its low-lying structures are found to have a common hexahedral (H(2)O)(20) shell, which is consistent with the experimental X-ray structure, but the position of the additional H(2)O (or the H(3)O(+) ion) in different low-lying structures is different, while the H(3)O(+) ion is situated at the center of the cage in the corresponding X-ray structure. The overall shape of the confined H(+)(H(2)O)(21) cluster is significantly different from that of the free cluster in the gas phase, and the confined cluster has much less intrinsic hydrogen bonds (H-bonds) than the free cluster, due to the need to form extrinsic H-bonds with acetonitrile molecules in the crystal host. The local electrostatic environment of the crystal host is found to exert a negligible influence on the structure of the (H(2)O)(21) cluster but play a significant role in modulating the structure of the H(+)(H(2)O)(21) cluster. This may be attributed to the fact that the protonated water cluster is much more easily polarized by the external electrostatic field of the crystal host than the neutral water cluster. PMID- 21634418 TI - Synthesis and photophysics of dibenz[a,c]phenazine derivatives. AB - The synthesis of dipolar dibenz[a,c]phenazine (DBP) derivatives is described. The compounds possess little electronic communication between donor and acceptor units in the ground state regardless of the pattern of substitution. The dipolar derivatives deactivate mostly via electron transfer (eT) under polar conditions. Intersystem crossing is likely to compete for S(1) relaxation. PMID- 21634419 TI - Design of chiral sulfoxide-olefins as a new class of sulfur-based olefin ligands for asymmetric catalysis. AB - The design and development of a novel class of chiral sulfur-olefin hybrid ligands with high synthetic feasibility are described. These new sulfoxide-olefin ligands showed excellent catalytic activities and enantioselectivities (up to 98% ee) in rhodium-catalyzed asymmetric 1,4-addition reactions of aryl boronic acids to alpha,beta-unsaturated carbonyl compounds. PMID- 21634420 TI - Synthesis and ring-chain-ring tautomerism of bisoxazolidines, thiazolidinyloxazolidines, and spirothiazolidines. AB - The synthesis of fused heterocycles such as thiazolidinyl-oxazolidine 3 is described starting from Tris.HCl. The mercaptomethyl bisoxazolidine 8 was found to convert to the corresponding thiazolidinyloxazolidine 3 and the spiro heterocycle 4 by a ring-chain-ring tautomerism, depending on the electronic nature of the ring substituents as well as the reaction conditions. This equilibration pathway is absent in the hydroxymethyl bisoxazolidines 2. Computational studies confirm that both kinetic and thermodynamic control features play a role in the product distribution. PMID- 21634421 TI - Cyclic nucleotide-gated channel block by hydrolysis-resistant tetracaine derivatives. AB - To meet a pressing need for better cyclic nucleotide-gated (CNG) channel antagonists, we have increased the biological stability of tetracaine-based blockers by synthesizing amide and thioamide linkage substitutions of tetracaine (1) and a higher affinity octyl tail derivative (5). We report the apparent K(D) values, the mechanism of block, and the in vitro hydrolysis rates for these compounds. The ester linkage substitutions did not adversely affect CNG channel block; unexpectedly, thioamide substitution in 1 (compound 8) improved block significantly. Furthermore, the ester linkage substitutions did not appear to affect the mechanism of block in terms of the strong state preference for closed channels. All ester substituted compounds, especially the thioamide substitutions, were more resistant to hydrolysis by serum cholinesterase than their ester counterparts. These findings have implications for dissecting the physiological roles of CNG channels, treating certain forms of retinal degeneration, and possibly the current clinical uses of compound 1. PMID- 21634422 TI - A [2 + 2] cycloaddition dimer and a Diels-Alder adduct from Alpinia katsumadai. AB - An unusual katsumadain dimer via a [2 + 2] cycloaddition, katsumadain C (1), and a unique chalcone-diarylheptanoid adduct via a Diels-Alder reaction, calyxin Y (2) with novel carbon frameworks, were isolated from the seeds of Alpinia katsumadai. Their structures and relative configurations were determined by spectroscopic evidence. PMID- 21634424 TI - Effects of sediment-associated extractable metals, degree of sediment grain sorting, and dissolved organic carbon upon Cryptosporidium parvum removal and transport within riverbank filtration sediments, Sonoma County, California. AB - Oocysts of the protozoan pathogen Cryptosporidium parvum are of particular concern for riverbank filtration (RBF) operations because of their persistence, ubiquity, and resistance to chlorine disinfection. At the Russian River RBF site (Sonoma County, CA), transport of C. parvum oocysts and oocyst-sized (3 MUm) carboxylate-modified microspheres through poorly sorted (sorting indices, sigma(1), up to 3.0) and geochemically heterogeneous sediments collected between 2 and 25 m below land surface (bls) were assessed. Removal was highly sensitive to variations in both the quantity of extractable metals (mainly Fe and Al) and degree of grain sorting. In flow-through columns, there was a log-linear relationship (r(2) = 0.82 at p < 0.002) between collision efficiency (alpha, the probability that colloidal collisions with grain surfaces would result in attachment) and extractable metals, and a linear relationship (r(2) = 0.99 at p < 0.002) between alpha and sigma(1). Collectively, variability in extractable metals and grain sorting accounted for ~83% of the variability in alpha (at p < 0.0002) along the depth profiles. Amendments of 2.2 mg L(-1) of Russian River dissolved organic carbon (DOC) reduced alpha for oocysts by 4-5 fold. The highly reactive hydrophobic organic acid (HPOA) fraction was particularly effective in re-entraining sediment-attached microspheres. However, the transport-enhancing effects of the riverine DOC did not appear to penetrate very deeply into the underlying sediments, judging from high alpha values (~1.0) observed for oocysts being advected through unamended sediments collected at ~2 m bls. This study suggests that in evaluating the efficacy of RBF operations to remove oocysts, it may be necessary to consider not only the geochemical nature and size distribution of the sediment grains, but also the degrees of sediment sorting and the concentration, reactivity, and penetration of the source water DOC. PMID- 21634425 TI - Alkaline-earth metal hydrides as novel host lattices for Eu(II) luminescence. AB - Luminescence of divalent europium has been investigated for the first time in metal hydrides. A complete solid-solution series was found for the pseudobinary system Eu(x)Sr(1-x)H(2) [a = 637.6(1) pm -12.1(3)x pm, b = 387.0(1)-6.5(2)x pm, c = 732.2(2)-10.1(4)x pm]. Europium-doped alkaline-earth hydrides Eu(x)M(1-x)H(2) (M = Ca, Sr, Ba) with a small europium concentration (x = 0.005) exhibit luminescence with maximum emission wavelengths of 764 nm (M = Ca), 728 nm (M = Sr), and 750 nm (M = Ba); i.e., the emission energy of divalent europium shows an extremely large red shift compared to the emission energies of fluorides or oxides. Theoretical calculations (LDA+U) confirm decreasing band gaps with increasing europium content of the solid solutions. PMID- 21634426 TI - Mixed-metal complexes incorporating platinum and lanthanide centers for selective binding and chirality sensing of succinates. AB - A chromophoric platinum complex was combined with a nonacoordinated cyclen lanthanide complex to give a new series of mixed-metal receptors. They specifically formed 1:1 complexes with dicarboxylates and offered selective chirality sensing of succinates. PMID- 21634427 TI - Comment on "Exothermic rate restrictions in long-range photoinduced charge separations". PMID- 21634428 TI - Comment on "Revised mechanism of Boyland-Sims oxidation". PMID- 21634429 TI - Chemical and biological evaluation of dipeptidyl boronic acid proteasome inhibitors for use in prodrugs and pro-soft drugs targeting solid tumors. AB - Bortezomib, a dipeptidyl boronic acid and potent inhibitor of the 26S proteasome, is remarkably effective against multiple myeloma (MM) but not against solid tumors. Dose-limiting adverse effects from "on target" inhibition of the proteasome in normal cells and tissues appear to be a key obstacle. Achieving efficacy against solid tumors therefore is likely to require making the inhibitor more selective for tumor tissue over normal tissues. The simplest strategy that might provide such tissue specificity would be to employ a tumor specific protease to release an inhibitor from a larger, noninhibitory structure. However, such release would necessarily generate an inhibitor with a free N-terminal amino group, raising a key question: Can short peptide boronic acids with N-terminal amino groups have the requisite properties to serve as warheads in prodrugs? Here we show that dipeptides of boroLeu, the smallest plausible candidates for the task, can indeed be sufficiently potent, cell-penetrating, cytotoxic, and stable to degradation by cellular peptidases to serve in this capacity. PMID- 21634431 TI - Electroluminescence from conjugated polymer electrospun nanofibers in solution processable organic light-emitting diodes. AB - Nanofibers of poly[(9,9-dioctylfluorenyl-2,7-diyl)-alt-co-(1,4-benzo-{2,1',3} thiadiazole)] (F8BT) blended in polystyrene (PS) or polyethylene oxide (PEO) show different diameters and morphology according to the conjugated polymer concentration. Electroluminescence from ribbonlike F8BT nanofibers, obtained by an annealing process of the F8BT/PEO blend, is successfully obtained by applying 6 V bias. Electrical connection is achieved by incorporating the F8BT fibers of about 700 nm width and 110 nm height into a single layer organic light emitting device, whose architecture induces charge recombination on the conjugated polymer nanofibers. This simple method to electrically connect the conjugated polymer nanofibers offers a great potential for low-cost flexible nanodevice fabrication. PMID- 21634430 TI - Discovery of (2E)-3-{2-butyl-1-[2-(diethylamino)ethyl]-1H-benzimidazol-5-yl}-N hydroxyacrylamide (SB939), an orally active histone deacetylase inhibitor with a superior preclinical profile. AB - A series of 3-(1,2-disubstituted-1H-benzimidazol-5-yl)-N-hydroxyacrylamides (1) were designed and synthesized as HDAC inhibitors. Extensive SARs have been established for in vitro potency (HDAC1 enzyme and COLO 205 cellular IC(50)), liver microsomal stability (t(1/2)), cytochrome P450 inhibitory (3A4 IC(50)), and clogP, among others. These parameters were fine-tuned by carefully adjusting the substituents at positions 1 and 2 of the benzimidazole ring. After comprehensive in vitro and in vivo profiling of the selected compounds, SB939 (3) was identified as a preclinical development candidate. 3 is a potent pan-HDAC inhibitor with excellent druglike properties, is highly efficacious in in vivo tumor models (HCT-116, PC-3, A2780, MV4-11, Ramos), and has high and dose proportional oral exposures and very good ADME, safety, and pharmaceutical properties. When orally dosed to tumor-bearing mice, 3 is enriched in tumor tissue which may contribute to its potent antitumor activity and prolonged duration of action. 3 is currently being tested in phase I and phase II clinical trials. PMID- 21634432 TI - Hydrogen-incorporation stabilization of metallic VO2(R) phase to room temperature, displaying promising low-temperature thermoelectric effect. AB - Regulation of electron-electron correlation has been found to be a new effective way to selectively control carrier concentration, which is a crucial step toward improving thermoelectric properties. The pure electronic behavior successfully stabilized the nonambient metallic VO(2)(R) to room temperature, giving excellent thermoelectric performance among the simple oxides with wider working temperature ranges. PMID- 21634433 TI - Protein disorder prevails under crowded conditions. AB - Crowding caused by the high concentrations of macromolecules in the living cell changes chemical equilibria, thus promoting aggregation and folding reactions of proteins. The possible magnitude of this effect is particularly important with respect to the physiological structure of intrinsically disordered proteins (IDPs), which are devoid of well-defined three-dimensional structures in vitro. To probe this effect, we have studied the structural state of three IDPs, alpha casein, MAP2c, and p21(Cip1), in the presence of the crowding agents Dextran and Ficoll 70 at concentrations up to 40%, and also the small-molecule osmolyte, trimethylamine N-oxide (TMAO), at concentrations up to 3.6 M. The structures of IDPs under highly diluted and crowded conditions were compared by a variety of techniques, fluorescence spectroscopy, acrylamide quenching, 1-anilino-8 naphthalenesulfonic acid (ANS) binding, fluorescence correlation spectroscopy (FCS), and far-UV and near-UV circular dichroism (CD) spectroscopy, which allow us to visualize various levels of structural organization within these proteins. We observed that crowding causes limited structural changes, which seem to reflect the functional requirements of these IDPs. alpha-Casein, a protein of nutrient function in milk, changes least under crowded conditions. On the other hand, MAP2c and, to a lesser degree, p21(Cip1), which carry out their functions by partner binding and accompanying partially induced folding, show signs of local structuring and also some global compaction upon crowded conditions, in particular in the presence of TMAO. The observations are compatible with the possible preformation of binding-competent conformations in these proteins. The magnitude of these changes, however, is far from that of the cooperative folding transitions elicited by crowding in denatured globular proteins; i.e., these IDPs do remain in a state of rapidly interconverting structural ensemble. Altogether, our results underline that structural disorder is the physiological state of these proteins. PMID- 21634434 TI - Synthesis and stabilization of subnanometric gold oxide nanoparticles on multiwalled carbon nanotubes and their catalytic activity. AB - Small gold nanoclusters in a very narrow size distribution (1.1 +/- 0.5 nm) have been stabilized onto multiwalled carbon nanotubes (MWCNT). Theoretical studies supported by XPS and (16)O(2)/(18)O(2) isotopic exchange experiments have shown that, on small gold nanoparticles (0.9-1.5 nm), dissociation of molecular O(2) and formation of a surface oxide-like layer is energetically favorable and occurs at room temperature, while O(2) recombination and desorption involves a larger activation barrier. CO titration experiments and theoretical studies demonstrate that the reactivity of the oxidized particles toward CO does not only depend on particle size but also on oxygen coverage. The oxidation-reduction process described is reversible, and the oxidized nanoparticles are active in the epoxidation of styrene with air. PMID- 21634435 TI - Insights into the early dissolution events of amlodipine using UV imaging and Raman spectroscopy. AB - Traditional dissolution testing determines drug release to the bulk, but does not enable an understanding of the events happening close to the surface of a solid or a tablet. UV imaging is a new imaging approach that can be used to study the dissolution behavior of chemical compounds. The UV imaging instrumentation offers recording of absorbance maps with a high spatial and temporal resolution which facilitates the abundant collection of information regarding the evolving solution concentrations. In this study, UV imaging was used to visualize the dissolution behavior of amlodipine besylate (amorphous and dihydrate forms) and amlodipine free base. The dissolution of amlodipine besylate was faster from the amorphous form than from the crystalline forms. The UV imaging investigations suggested that a solvent mediated phase transformation occurred for the amorphous amlodipine besylate and the amlodipine free base samples. Raman spectroscopy was used to confirm and probe the changes at the solid surface occurring upon contact with the dissolution media and verified the recrystallization of the amorphous form to the monohydrate. The combination of UV imaging and Raman spectroscopy is an efficient tool to obtain a deeper insight into the early events of the dissolution process. PMID- 21634436 TI - Liposomal drug formulations in the treatment of rheumatoid arthritis. AB - Liposomes have been extensively investigated as drug delivery systems in the treatment of rheumatoid arthritis (RA). Low bioavailability, high clearance rates and limited selectivity of several important drugs used for RA treatment require high and frequent dosing to achieve sufficient therapeutic efficacy. However, high doses also increase the risk for systemic side effects. The use of liposomes as drug carriers may increase the therapeutic index of these antirheumatic drugs. Liposomal physicochemical properties can be changed to optimize penetration through biological barriers and retention at the site of administration, and to prevent premature degradation and toxicity to nontarget tissues. Optimal liposomal properties depend on the administration route: large-sized liposomes show good retention upon local injection, small-sized liposomes are better suited to achieve passive targeting. PEGylation reduces the uptake of the liposomes by liver and spleen, and increases the circulation time, resulting in increased localization at the inflamed site due to the enhanced permeability and retention (EPR) effect. Additionally liposomal surfaces can be modified to achieve selective delivery of the encapsulated drug to specific target cells in RA. This review gives an overview of liposomal drug formulations studied in a preclinical setting as well as in clinical practice. It covers the use of liposomes for existing antirheumatic drugs as well as for new possible treatment strategies for RA. Both local administration of liposomal depot formulations and intravenous administration of passively and actively targeted liposomes are reviewed. PMID- 21634437 TI - Peptoid atropisomers. AB - We report the isolation of N-aryl peptoid oligomers that adopt chiral folds, despite the absence of chiral centers. Peptoid monomers incorporating ortho substituted N-aryl side chains are identified that exhibit axial chirality. We observe significant energy barriers to rotation about the stereogenic carbon nitrogen bond, allowing chromatographic purification of stable atropisomeric forms. We study the atropisomerism of N-aryl peptoid oligomers by computational modeling, NMR, X-ray crystallography, dynamic HPLC, and circular dichroism. The results demonstrate a new approach to promote the conformational ordering of this important class of foldamer compounds. PMID- 21634438 TI - Use of the dual potential to rationalize the occurrence of some DNA lesions (pyrimidic dimers). AB - Exploiting the locality of the chemical potential of an excited state when it is evaluated using the ground state Density Functional Theory (DFT), a new local descriptor for excited states has been proposed (J. Chem. Theory Comput.2009, 5, 2274). This index is based on the assumption that the relaxation of the electronic density toward that of the ground state drives the chemical reactivity of excited states. The sign of the descriptor characterizes the electrophilic or nucleophilic behavior of atomic regions. Through an exact excited state DFT formalism provided by Gross, Oliveira, and Kohn, a mathematical argument is given for this descriptor only for the first excited state. It is afterward used to rationalize the occurrence and the regioselectivity of some DNA lesions based on the [2 + 2] cycloaddition between two adjacent bases. PMID- 21634439 TI - Bottom-up shotgun lipidomics by higher energy collisional dissociation on LTQ Orbitrap mass spectrometers. AB - Higher energy collision dissociation (HCD) is a complementary fragmentation tool that has recently become available on mass spectrometers of the LTQ Orbitrap family. We report on a shotgun bottom-up lipidomics approach that relies on HCD of the isolated lipid precursors. HCD, together with the high mass resolution and mass accuracy of the Orbitrap analyzer, improved the confidence of molecular species assignment and accuracy of their quantification in total lipid extracts. These capabilities were particularly important for accounting for biologically interesting lipid species comprising polyunsaturated and odd numbered fatty acid moieties. We argue that now both bottom-up and top-down shotgun lipidomics could be performed on the same instrumentation platform. PMID- 21634649 TI - Other/miscellaneous. PMID- 21634440 TI - Time-dependent oxidation during nano-assisted laser desorption ionization mass spectrometry: a useful tool for structure determination or a source of possible confusion? AB - This work reports on a new and extremely simple approach for determination of a double bond position by a laser desorption ionization mass spectrometry. It is solely based on the catalytic properties of nanostructured surfaces used in nanoassisted laser desorption ionization experiments. These surfaces can induce oxidation of analytes, which results in a mass shift that can be detected by mass spectrometry. If a site of unsaturation is oxidized and cleaved, the m/z difference is diagnostic of the position of a double bond. By demonstrating that the oxidation depends on the analyte surface dwell time, it was proven that it is caused by the surface activity and not by the laser desorption ionization process itself. Control matrix-assisted laser desorption/ionization (MALDI) experiment showed only a limited partial oxidation and no time dependency of the process. The ability to determine a position of a double bond was demonstrated on polyunsaturated phospholipids and cyclosporine A. In some other cases, however, the unexpected oxidation could cause confusion, as demonstrated for a glycosphingolipid from a porcine brain extract. PMID- 21634651 TI - Other/miscellaneous. PMID- 21634650 TI - The Human Genome Project--and Beyond. PMID- 21634653 TI - In AC Research: In AC Research. PMID- 21634655 TI - Analytical Currents: An affinity for epitopes. PMID- 21634654 TI - Analytical Currents: Different neurons, differing peptides. PMID- 21634656 TI - Analytical Currents: SERS gets the fat. PMID- 21634657 TI - Analytical Currents: Chromatographic dispersion by NMR. PMID- 21634658 TI - Analytical Currents: Single-shot analysis. PMID- 21634660 TI - Analytical Currents: Let the gold fly. PMID- 21634659 TI - Analytical Currents: Look out peptides, chemosensors are here. PMID- 21634661 TI - Analytical Currents: Inside a single vesicle. PMID- 21634662 TI - Analytical Currents: Electrochemical immunosensor targets PCBs. PMID- 21634663 TI - Analytical Currents: Separating structurally diverse combinatorial mixtures. PMID- 21634665 TI - Science: Field analysts keep up the pace. PMID- 21634664 TI - Science: Emerging analytical tests for prion proteins. PMID- 21634666 TI - Laboratory Profile: Proteomics unleashed. PMID- 21634667 TI - Science: Chemical sensing down the (fiber) line. PMID- 21634668 TI - Laboratory Profile: Beastly robots and glowing microscopes. PMID- 21634669 TI - News from Pittcon: Big plans for 50th. AB - The staff of Analytical Chemistry reports from New Orleans, LA. PMID- 21634671 TI - News from Pittcon: Beckman hopes to create protocol standard. AB - The staff of Analytical Chemistry reports from New Orleans, LA. PMID- 21634670 TI - News from Pittcon: Building a roadmap for industry. AB - The staff of Analytical Chemistry reports from New Orleans, LA. PMID- 21634672 TI - News from ABRF '98: Edman and MS go hand in hand. PMID- 21634674 TI - People: 1998 EAS award winners. PMID- 21634673 TI - People: velmer a. Fassel, 1919-98. PMID- 21634675 TI - Government and Society: International MS society moves forward. PMID- 21634678 TI - Peer Reviewed: Liquid chromatography as a measurement tool for chiral interactions. AB - Thermodynamic measurements using LC demonstrate the breadth of this technique. PMID- 21634676 TI - Business: HP acquires MTI. PMID- 21634679 TI - Peer reviewed: nanomaterials in analytical chemistry. AB - As particle size approaches molecular dimensions, all properties of a material change, making nanomaterials useful for particular applications. PMID- 21634681 TI - The barnett institute at 25. AB - The Barnett Institute has established a world-class reputation and positioned itself well for the future. PMID- 21634683 TI - Meetings: conferences. PMID- 21634682 TI - Product Review: Fattening up SFE sales. AB - Fat-labeling requirements place hefty demand on extraction market. PMID- 21634685 TI - Meetings: short courses. PMID- 21634684 TI - Meetings: ASMS in Orlando. PMID- 21634686 TI - A: the sound of talking. AB - all which isn't talking is mere typing and all typing's typing to oneself but the very tone of talking is telling. PMID- 21634688 TI - Books: Recruiting CE users. PMID- 21634689 TI - Books: More than a cookbook. AB - A review of Instrumental Methods in Food Analysis. PMID- 21634690 TI - Books: One molecule at a time. AB - A review of Single-Molecule Optical Detection, Imaging, and Spectroscopy. PMID- 21634692 TI - New products: new products. PMID- 21634694 TI - Other/miscellaneous. PMID- 21634695 TI - Other/miscellaneous. PMID- 21634787 TI - Amyloid of the Candida albicans Ure2p prion domain is infectious and has an in register parallel beta-sheet structure. AB - Ure2p of Candida albicans (Ure2(albicans) or CaUre2p) can be a prion in Saccharomyces cerevisiae, but Ure2p of Candida glabrata (Ure2(glabrata)) cannot, even though the Ure2(glabrata) N-terminal domain is more similar to that of the S. cerevisiae Ure2p (Ure2(cerevisiae)) than Ure2(albicans) is. We show that the N terminal N/Q-rich prion domain of Ure2(albicans) forms amyloid that is infectious, transmitting [URE3alb] to S. cerevisiae cells expressing only C. albicans Ure2p. Using solid-state nuclear magnetic resonance of selectively labeled C. albicans Ure2p(1-90), we show that this infectious amyloid has an in register parallel beta-sheet structure, like that of the S. cerevisiae Ure2p prion domain and other S. cerevisiae prion amyloids. In contrast, the N/Q-rich N terminal domain of Ure2(glabrata) does not readily form amyloid, and that formed upon prolonged incubation is not infectious. PMID- 21634788 TI - Tetrahedral atom ordering in a zeolite framework: a key factor affecting its physicochemical properties. AB - Three gallosilicate natrolites with closely similar chemical composition but differing in the distribution of Si and Ga over crystallographically different tetrahedral sites (T-sites) show striking differences in their cation exchange performance. The ability to exchange Na(+) by the larger alkali metal cations decreases upon increasing the size of the cation, as expected, but also with the degree of T-atom ordering. To seek an insight into this phenomenon, the crystal structures of 11 different zeolites, which show variations in degree of T-atom ordering, nature of countercation, and hydration state, have been refined using synchrotron diffraction data. While the three as-made sodium materials were characterized to have a low, medium, and high degree of ordering, respectively, their pore sizes are close to the size of the bare Na(+) cation and much smaller than that of the larger alkali cations, which are nonetheless exchanged into the materials, each one at a different level. Interestingly, large differences are also manifested when the Na(+) back-exchange is performed on the dehydrated K(+) forms, with crystallographic pore sizes too small even to allow the passage of Na(+). Although the thermodynamic data point to small differences in the enthalpy of the Na(+)/K(+) exchange in the three materials, comparison of the "static" crystallographic pore sizes and the diameter of the exchanged cations lead us to conclude that during the exchange process these zeolites undergo significant deformations that dynamically open the pores, allowing cation traffic even for Cs(+) in the case of the most disordered material. In addition to the very large topological flexibility typical of the natrolite framework, we propose as a hypothesis that there is an additional flexibility mechanism that decreases the rigidity of the natrolite chain itself and is dependent on preferential siting of Si or Ga on crystallographically different T-sites. PMID- 21634789 TI - Structure of the response regulator PhoP from Mycobacterium tuberculosis reveals a dimer through the receiver domain. AB - The PhoP protein from Mycobacterium tuberculosis is a response regulator of the OmpR/PhoB subfamily, whose structure consists of an N-terminal receiver domain and a C-terminal DNA-binding domain. How the DNA-binding activities are regulated by phosphorylation of the receiver domain remains unclear due to a lack of structural information on the full-length proteins. Here we report the crystal structure of the full-length PhoP of M. tuberculosis. Unlike other known structures of full-length proteins of the same subfamily, PhoP forms a dimer through its receiver domain with the dimer interface involving alpha4-beta5 alpha5, a common interface for activated receiver domain dimers. However, the switch residues, Thr99 and Tyr118, are in a conformation resembling those of nonactivated receiver domains. The Tyr118 side chain is involved in the dimer interface interactions. The receiver domain is tethered to the DNA-binding domain through a flexible linker and does not impose structural constraints on the DNA binding domain. This structure suggests that phosphorylation likely facilitates/stabilizes receiver domain dimerization, bringing the DNA-binding domains to close proximity, thereby increasing their binding affinity for direct repeat DNA sequences. PMID- 21634790 TI - Recyclable SERS substrates based on Au-coated ZnO nanorods. AB - Vertically aligned Au-coated ZnO nanorods (Au-ZnO NRs) were investigated as cheap, efficient and recyclable SERS-active substrates. The ZnO NRs were prepared through a simple, low-temperature hydrothermal route and made SERS-active through deposition of gold nanoislands by sputtering at room temperature. Optimized samples were able to detect methylene blue over a wide range of low concentrations (from 1 * 10(-4) to 1 * 10(-12) M), with good reproducibility. The photocatalytic properties of Au-ZnO NRs were exploited to recycle these substrates through UV-assisted cleaning. The experimental results showed that these substrates are characterized by high reproducibility and long shelf life, which make them promising as SERS platforms for multiple detection of different molecular species. PMID- 21634791 TI - Self-assembly of focal point oligo-catechol ethylene glycol dendrons on titanium oxide surfaces: adsorption kinetics, surface characterization, and nonfouling properties. AB - This work covers the synthesis of second-generation, ethylene glycol dendrons covalently linked to a surface anchor that contains two, three, or four catechol groups, the molecular assembly in aqueous buffer on titanium oxide surfaces, and the evaluation of the resistance of the monomolecular adlayers against nonspecific protein adsorption in contact with full blood serum. The results were compared to those of a linear poly(ethylene glycol) (PEG) analogue with the same molecular weight. The adsorption kinetics as well as resulting surface coverages were monitored by ex situ spectroscopic ellipsometry (VASE), in situ optical waveguide lightmode spectroscopy (OWLS), and quartz crystal microbalance with dissipation (QCM-D) investigations. The expected compositions of the macromolecular films were verified by X-ray photoelectron spectroscopy (XPS). The results of the adsorption study, performed in a high ionic strength ("cloud point") buffer at room temperature, demonstrate that the adsorption kinetics increase with increasing number of catechol binding moieties and exceed the values found for the linear PEG analogue. This is attributed to the comparatively smaller and more confined molecular volume of the dendritic macromolecules in solution, the improved presentation of the catechol anchor, and/or their much lower cloud-point in the chosen buffer (close to room temperature). Interestingly, in terms of mechanistic aspects of "nonfouling" surface properties, the dendron films were found to be much stiffer and considerably less hydrated in comparison to the linear PEG brush surface, closer in their physicochemical properties to oligo(ethylene glycol) alkanethiol self-assembled monolayers than to conventional brush surfaces. Despite these differences, both types of polymer architectures at saturation coverage proved to be highly resistant toward protein adsorption. Although associated with higher synthesis costs, dendritic macromolecules are considered to be an attractive alternative to linear polymers for surface (bio)functionalization in view of their spontaneous formation of ultrathin, confluent, and nonfouling monolayers at room temperature and their outstanding ability to present functional ligands (coupled to the termini of the dendritic structure) at high surface densities. PMID- 21634792 TI - Oxidation of Au by surface OH: nucleation and electronic structure of gold on hydroxylated MgO(001). AB - The nucleation and electronic structure of vapor-deposited Au on hydroxylated MgO(001) surfaces has been investigated under ultrahigh vacuum conditions. Hydroxylated MgO(001) surfaces with two different hydroxyl coverages, 0.4 and 1 monolayer, respectively, were prepared by exposure to water (D(2)O) at room temperature. Scanning tunneling microscopy experiments show significantly higher gold particle densities and smaller particle sizes on the hydroxylated MgO surface as compared to gold deposited on clean MgO(001). Infrared spectroscopy and X-ray photoelectron spectroscopy experiments were performed to reveal details about the initial nucleation of gold. Gold atoms are found to chemically interact with a specific type of hydroxyl groups on the MgO surface, leading to the formation of oxidized gold particles. The enhanced adhesion of Au particles, which is due to the formation of strong Au-O interfacial bonds, is responsible for the observed higher stability of small Au clusters toward thermal sintering on hydroxylated MgO surfaces. The results are compared to similar studies on Au/TiO(2)(110) model systems and powder samples prepared by the deposition precipitation route. PMID- 21634793 TI - Chain-growth cycloaddition polymerization via a catalytic alkyne [2 + 2 + 2] cyclotrimerization reaction and its application to one-shot spontaneous block copolymerization. AB - A cobalt-catalyzed alkyne [2 + 2 + 2] cycloaddition reaction has been applied to polymerizations yielding linear polymers via selective cross-cyclotrimerization of yne-diyne monomers, which occurs in a chain-growth manner. Additionally, through control of the alkyne reactivity of the two monomers, this method was efficiently applied to the spontaneous block copolymerization of their mixture. Here we present the proposed mechanism of the catalyst transfer process of this cycloaddition polymerization. PMID- 21634794 TI - C-H functionalization polycondensation of chlorothiophenes in the presence of nickel catalyst with stoichiometric or catalytically generated magnesium amide. AB - Polymerization of 2-chloro-3-substituted thiophenes proceeded with a stoichiometric amount of magnesium amide, TMPMgCl.LiCl, or a combination of a Grignard reagent and a catalytic amount of secondary amine in the presence of a nickel catalyst. Although the nickel-catalyzed polymerization with NiCl(2)dppe, which exhibited high catalytic activity in the reaction of bromothiophenes, was less effective, use of a nickel catalyst bearing N-heterocyclic carbene as a ligand was found to induce polymerization with controlled molecular weight and molecular weight distribution. PMID- 21634795 TI - Regional cooperativity in the phase transitions of dipalmitoylphosphatidylcholine bilayers: the lipid tail triggers the isothermal crystallization process. AB - We have a long-standing interest to explore the answer of the question: Which part of the amphiphilic molecule triggers the phase transition of the self assembled aggregates consisting of these amphiphiles? This is an important issue regarding the phase transition kinetics of amphiphiles. To this end, we studied the phase transition behaviors of dipalmitoylphosphatidylcholine (DPPC) by differential scanning calorimetry, synchrotron X-ray scattering, Fourier transform infrared spectroscopy, and image analysis. We found that different parts (head, interface, and tail) of DPPC molecules all exhibit nonsynchronous changes during the sub-, pre-, and main transitions. Particular efforts have been devoted to studying the isothermal subgel (L(c')) formation process. It was found that only the lipid interface and tail regions change, and only when the rearrangement of the lipid hydrocarbon chain packing reaches a certain extent can the interfacial C?O groups be induced to undergo vibrational environment changes. The result means that the hydrocarbon tail is the part that triggers the gel (L(beta')) to L(c') phase transition. The present work deepens our understanding on the phase transition mechanisms of DPPC and may shed light on those of other phospholipids and other types of amphiphiles. PMID- 21634796 TI - Formation of through-holes in Si wafers by using anodically polarized needle electrodes in HF solution. AB - Electrochemical pore formation in Si using an anodized needle electrode was studied. In the electrochemical process, a Pt, Ir or Pd needle with a diameter of 50-200 MUm was brought into contact at its tip with a Si wafer, which was not connected to an external circuit, in HF solution. By applying an anodic potential to the needle electrode against a Pt counter electrode, a pore with a diameter slightly larger than the diameter of the needle electrode was formed in both p type and n-type Si, of which current efficiency was higher for n-type Si. Through holes were electrochemically formed in p-type and n-type Si wafers at speeds higher than 30 MUm min(-1) using a sharpened Ir needle electrode. A model was proposed to explain the results, in which the pore formation was attributed to successive dissolution of Si atoms near the 3-phase (Si/metal/HF solution) boundary by positive holes injected from the needle electrode to the surface of Si. PMID- 21634797 TI - Assembly of lipid bilayers on silica and modified silica colloids by reconstitution of dried lipid films. AB - A method is presented for the assembly of lipid bilayers on silica colloids via reconstitution of dried lipid films solvent-cast from chloroform within packed beds of colloids ranging from 100 nm to 10 MUm in diameter. Rapid solvent evaporation from the packed bed void volume results in uniform distribution of dried lipid throughout the colloidal bed. Fluorescence measurements indicate that significant, if not quantitative, retention of DOPC or DPPC films cast between sub-bilayer and multilayer quantities occurs when the colloids are redispersed in aqueous solution. Phospholipid bilayers assembled in this manner are shown to effectively passivate the surface of 250 nm colloids to nonspecific adsorption of bovine serum albumin. The method is shown to be capable of preparing supported bilayers on colloid surfaces that do not generally support vesicle fusion such as poly(ethylene glycol) (PEG) modified silica colloids. Bilayers of lipids that have not been reported to self-assemble by vesicle fusion, including gel-phase lipids and single-chain diacetylene amphiphiles, can also be formed by this method. The utility of the solid-core support is demonstrated by the facile assembly of supported lipid bilayers within fused silica capillaries to generate materials that are potentially suitable for the analysis of membrane interactions in a microchannel format. PMID- 21634798 TI - Boronic acid hydrogen bonding in encapsulation complexes. AB - Hydrogen bonding is a key determinant of much macromolecular structure in nature, but individual donor and acceptor pairs are rarely observed in solution. Their weak interactions result in nanosecond lifetimes and rapid exchange of partners. Reversible encapsulation isolates molecules in very small spaces for milliseconds to hours and allows their characterization by NMR methods. Here we report a competitive study of hydrogen-bonding functions--carboxylic acids, primary amides, and boronic acids--within a multicomponent capsular assembly. The pairwise co-encapsulation of these molecules allows the direct observation of homodimeric boronic acids and their heterodimeric complexes with carboxylic acids and primary amides. The efficiency of boronic acids as hydrogen-bonding partners derives from their adaptable structures rather than from their intrinsic acid/base properties. PMID- 21634799 TI - Production of 8-prenylnaringenin from isoxanthohumol through biotransformation by fungi cells. AB - 8-Prenylnaringenin (8PN), which presents in hop, enjoys fame as the most potential phytoestrogen. Although a number of health effects are attributed to 8PN, few reports are available about the production of it. In this work, screening of fungi to efficiently transform isoxanthohumol (IXN) into 8PN was designed. The biotransformation of IXN was significantly observed in Eupenicillium javanicum, Cunninghamella blakesleana, and Ceriporiopsis subvermispora under five kinds of transformation conditions. As a comparative result of IXN transformation, E. javanicum was the optimal biocatalyst to produce 8PN. Transformation caused by growing precultured fungal mycelia, a process designated as G2, was a favorable condition for IXN transformation in view of the yield of 8PN. The possible transformation pathway of 8PN bioproduction is postulated in this work. The construction of fungus and transformation mode derived from the current work is viable and an alternative procedure for 8PN formation. PMID- 21634801 TI - Study of defect sites in Ce1-xMxO2-delta (x = 0.2) solid solutions using Raman spectroscopy. AB - A series of Ce(1-x)M(x)O(2-delta) (M = Gd, Zr, La, Sm, Y, Lu, and Pr) samples were characterized by Raman spectroscopy to investigate the evolution of defect sites (oxygen vacancies and MO(8)-type complex) and their distributions in the samples. It was found that the evolution of oxygen vacancies was due to the different ionic valence state of dopant from that of Ce(4+), while the evolution of the MO(8)-type complex was due to the different ionic radius of dopant from that of Ce(4+). The distributions of defect sites were investigated using 325 and 514 nm excitation laser lines, indicating that the defect sites were surface enriched. Moreover, the increasing ordering level of the sample led to a decline in the concentration of the MO(8)-type complex in the sample but the constant concentration of oxygen vacancies, implying that the metastable MO(8)-type complex species were more disordered compared to the oxygen vacancies. PMID- 21634800 TI - Synthesis of folate-functionalized RAFT polymers for targeted siRNA delivery. AB - Receptor-mediated, cell-specific delivery of siRNA enables silencing of target genes in specific tissues, opening the door to powerful therapeutic options for a multitude of diseases. However, the development of delivery systems capable of targeted and effective siRNA delivery typically requires multiple steps and the use of sophisticated, orthogonal chemistries. Previously, we developed diblock copolymers consisting of dimethaminoethyl methacrylate-b-dimethylaminoethyl methacrylate-co-butyl methacrylate-co-propylacrylic acid as potent siRNA delivery systems that protect siRNA from enzymatic degradation and enable its cytosolic delivery through pH-responsive, endosomolytic behavior. (1, 2) These architectures were polymerized using a living radical polymerization method, specifically reversible addition-fragmentation chain transfer (RAFT) polymerization, which employs a chain transfer agent (CTA) to modulate the rate of reaction, resulting in polymers with low polydispersity and telechelic chain ends reflecting the chemistry of the CTA. Here we describe the straightforward, facile synthesis of a folate receptor-targeted diblock copolymer siRNA delivery system because the folate receptor is an attractive target for tumor-selective therapies as a result of its overexpression in a number of cancers. Specifically, we detail the de novo synthesis of a folate-functionalized CTA, use the folate CTA for controlled polymerizations of diblock copolymers, and demonstrate efficient, specific cellular folate receptor interaction and in vitro gene knockdown using the folate-functionalized polymer. PMID- 21634802 TI - Toward label-free optical fractionation of blood--optical force measurements of blood cells. AB - There is a compelling need to develop systems capable of processing blood and other particle streams for detection of pathogens that are sensitive, selective, automated, and cost/size effective. Our research seeks to develop laser-based separations that do not rely on prior knowledge, antibodies, or fluorescent molecules for pathogen detection. Rather, we aim to harness inherent differences in optical pressure, which arise from variations in particle size, shape, refractive index, or morphology, as a means of separating and characterizing particles. Our method for measuring optical pressure involves focusing a laser into a fluid flowing opposite to the direction of laser propagation. As microscopic particles in the flow path encounter the beam, they are trapped axially along the beam and are pushed upstream from the laser focal point to rest at a point where the optical and fluid forces on the particle balance. On the basis of the flow rate at which this balance occurs, the optical pressure felt by the particle can be calculated. As a first step in the development of a label free device for processing blood, a system has been developed to measure optical pressure differences between the components of human blood, including erythrocytes, monocytes, granulocytes, and lymphocytes. Force differentials have been measured between various components, indicating the potential for laser based separation of blood components based upon differences in optical pressure. Potential future applications include the early detection of blood-borne pathogens for the prevention of sepsis and other diseases as well as the detection of biological threat agents. PMID- 21634803 TI - Poly[N-(2-hydroxypropyl)methacrylamide]-based tissue-embedding medium compatible with MALDI mass spectrometry imaging experiments. AB - Traditional tissue-sectioning techniques for histological samples utilize various embedding media to stabilize the tissue on a sectioning target and to provide a smooth cutting surface. Due to the ion suppression effect in MALDI ionization and number of background peaks in the low-mass region, these media are not suitable for mass spectrometry imaging (MSI) experiments. To overcome this, droplets of water are often used to mount the tissue on a sectioning target, but the ice block formed around the tissue does not provide a good support for sectioning of fragile samples. In this work, we propose a novel embedding media, compatible with MALDI ionization and MSI experiments, based on poly[N-(2 hydroxypropyl)methacrylamide] (pHPMA). Using a reversible addition-fragmentation chain transfer polymerization technique, well-defined pHPMA polymer with narrow mass distribution was prepared. Benefits of the resulted pHPMA-based embedding media were tested on different tissue samples. PMID- 21634804 TI - Visualization of nonengineered single mRNAs in living cells using genetically encoded fluorescent probes. AB - Single mRNA imaging in live cells is a useful technique to elucidate its precise localization and dynamics. We developed a method for visualizing endogenous mRNAs in living cells with single molecule sensitivity using genetically encoded probes. An RNA-binding protein of human PUMILIO1 (PUM-HD) was used for recognizing base sequences of a target mRNA, beta-actin mRNA. Two PUM-HDs were modified by amino acid mutations to bind specifically to tandem 8-base sequences of the target mRNA. Because each PUM-HD was connected with amino- and carboxyl terminal fragments of enhanced green fluorescent protein (EGFP), the probes emit fluorescence by reconstitution of EGFP fragments upon binding to beta-actin mRNAs. The EGFP reconstituted on the mRNAs was monitored with a total internal reflection fluorescence microscope. Results show that each fluorescent spot in live cells represented a single beta-actin mRNA and that distinct spatial and temporal movement of the individual beta-actin mRNAs was visualized. We also estimated the average velocity of the movement of the single mRNAs along microtubules in live cells. This method is widely applicable to tracking various mRNAs of interest in the native state of living cells with single-mRNA sensitivity. PMID- 21635006 TI - A reproducible and high-throughput HPLC/MS method to separate sarcosine from alpha- and beta-alanine and to quantify sarcosine in human serum and urine. AB - While sarcosine was recently identified as a potential urine biomarker for prostate cancer, further studies have cast doubt on its utility to diagnose this condition. The inconsistent results may be due to the fact that alanine and sarcosine coelute on an HPLC reversed-phase column and the mass spectrometer cannot differentiate between the two isomers, since the same parent/product ions are generally used to measure them. In this study, we developed a high-throughput liquid chromatography-mass spectrometry (LC-MS) method that resolves sarcosine from alanine isomers, allowing its accurate quantification in human serum and urine. Assay reproducibility was determined using the coefficient of variation (CV) and intraclass correlation coefficient (ICC) in serum aliquots from 10 subjects and urine aliquots from 20 subjects across multiple analytic runs. Paired serum/urine samples from 42 subjects were used to evaluate sarcosine serum/urine correlation. Both urine and serum assays gave high sensitivity (limit of quantitation of 5 ng/mL) and reproducibility (serum assay, intra- and interassay CVs < 3% and ICCs > 99%; urine assay, intra-assay CV = 7.7% and ICC = 98.2% and interassay CV = 12.3% and ICC = 94.2%). In conclusion, this high throughput LC-MS method is able to resolve sarcosine from alpha- and beta-alanine and is useful for quantifying sarcosine in serum and urine samples. PMID- 21635007 TI - Conformer-specific hydrogen exchange analysis of Abeta(1-42) oligomers by top down electron capture dissociation mass spectrometry. AB - Protein structural studies are particularly challenging under conditions in which several conformational species (e.g., monomers and aggregated forms) coexist in solution. Most spectroscopic techniques provide population-averaged data. Hence, it is usually not possible to obtain detailed structural information on individual protein species in heterogeneous samples. The current work employs an experimental strategy that addresses this issue. Solution-phase hydrogen exchange (HX) is used in combination with tandem mass spectrometry. Electrosprayed intact ions exhibiting specific HX mass shifts are selected in the gas phase, followed by electron capture dissociation. The resulting fragment ion deuteration pattern provides amide hydrogen bonding information in a conformer-specific and spatially resolved fashion. The feasibility of this approach is demonstrated by applying it to neurotoxic Abeta(1-42) oligomers that coexist with disordered monomers in solution. The findings of this study point to similarities between oligomers and mature amyloid fibrils with regard to the Abeta(1-42) backbone organization. Specifically, fibrils and oligomers appear to share a beta-loop-beta secondary structure motif. The spatial resolution obtained with the "top-down" approach used here exceeds that of earlier proteolysis-based HX data on Abeta. PMID- 21635008 TI - Time-temperature superposition and the controlling role of solvation in the viscoelastic properties of polyaniline thin films. AB - Comprehensive exploration of the viscoelastic properties of polyaniline films exposed to aqueous perchloric acid has been made as a function of applied potential (E), temperature (T), and mechanical oscillation frequency (f = omega/2pi) using high-frequency acoustic wave resonators. The outcomes are expressed in terms of storage and loss shear modulus signatures, G'(E, T, omega) and G"(E, T, omega). Surprisingly, these are barely sensitive to potential, through which both polymer charge and solvation are manipulated, and only modestly sensitive to temperature. In contrast, the response to timescale is dramatic. Using the principle of time-temperature superposition, G' and G" at different temperatures and frequencies (time scales) can each be placed on master relaxation curves. Models developed for mechanical properties of bulk polymers at low frequency were applied to these thin film responses at high frequency. These include the Williams-Landel-Ferry model, the activation model, and the Rouse-Zimm model based, respectively, on concepts of free volume, thermal activation, and relaxation. Each of the models could be applied with physically reasonable outcomes in terms of the relevant parameters (thermal expansion coefficient, glass transition temperature, and activation enthalpy). G' and G" values are correlated with solvent content. The enthalpy change for solvent entry is small, positive and relatively independent of polymer charge state, all of which contrast sharply with the behavior of thiophene-based conducting polymers in organic solvents. PMID- 21635009 TI - Identification of cyanidin 3-O-beta-(6"-(3-hydroxy-3-methylglutaroyl)glucoside) and other anthocyanins from wild and cultivated blackberries. AB - Anthocyanins from blackberries are natural dietary pigments. The aim of this study was to investigate the occurrence of anthocyanins in fruits of wild Norwegian blackberries and three blackberry ( Rubus fruticosus L.) cultivars and to report the complete identification of cyanidin 3-O-beta-(6"-(3-hydroxy-3 methylglutaroyl)glucopyranoside), 5. This new pigment is most probably the same pigment that has previously been reported to occur in various blackberry samples as cyanidin 3-dioxalylglucoside. All of the examined blackberry samples contained in similar relative proportions the 3-glucoside (1), 3-rutinoside (2), 3-xyloside (3), and 3-O-beta-(6"-malonylglucoside) (4) of cyanidin and 5. The absolute amounts of 1-5 in the wild Norwegian blackberries were 249, 18, 10, 24, and 22 mg of cyanidin 3-glucoside equivalents/100 g of fresh weight, respectively. PMID- 21635010 TI - From atomistic modeling to excitation transfer and two-dimensional spectra of the FMO light-harvesting complex. AB - The experimental observation of long-lived quantum coherences in the Fenna Matthews-Olson (FMO) light-harvesting complex at low temperatures has challenged general intuition in the field of complex molecular systems and provoked considerable theoretical effort in search of explanations. Here we report on room temperature calculations of the excited-state dynamics in FMO using a combination of molecular dynamics simulations and electronic structure calculations. Thus we obtain trajectories for the Hamiltonian of this system which contains time dependent vertical excitation energies of the individual bacteriochlorophyll molecules and their mutual electronic couplings. The distribution of energies and couplings is analyzed together with possible spatial correlations. It is found that in contrast to frequent assumptions the site energy distribution is non Gaussian. In a subsequent step, averaged wave packet dynamics is used to determine the exciton dynamics in the system. Finally, with the time-dependent Hamiltonian, linear and two-dimensional spectra are determined. The thus-obtained linear absorption line shape agrees well with experimental observation and is largely determined by the non-Gaussian site energy distribution. The two dimensional spectra are in line with what one would expect by extrapolation of the experimental observations at lower temperatures and indicate almost total loss of long-lived coherences. PMID- 21635011 TI - Synthesis, characterization, and electrochemical properties of a series of sterically varied iron(II) alkoxide precursors and their resultant nanoparticles. AB - A new family of iron(II) aryloxide [Fe(OAr)(2)(py)(x)] precursors was synthesized from the alcoholysis of iron(II) mesityl [Fe(Mes)(2)] in pyridine (py) using a series of sterically varied 2-alkyl phenols (alkyl = methyl (H-oMP), isopropyl (H oPP), tert-butyl (H-oBP)) and 2,6-dialkyl phenols (alkyl = methyl (H-DMP), isopropyl (H-DIP), tert-butyl (H-DBP), phenyl (H-DPhP)). All of the products were found to be mononuclear and structurally characterized as [Fe(OAr)(2)(py)(x)] (x = 3 OAr = oMP (1), oPP (2), oBP (3), DMP (4), DIP (5); x = 2 OAr = DBP (6), DPhP (7)). The use of tris-tert-butoxysilanol (OSi(OBu(t))(3) = TOBS) led to isolation of [Fe(TOBS)(2)(py)(2)] (8). The new Fe(OAr)(2)(py)(x) (1-6) were found, under solvothermal conditions, to produce nanodots identified by PXRD as the gamma maghemite phase. The model precursor 3 and the nanoparticles 6n were evaluated using electrochemical methods. Cyclic voltammetry for 3 revealed multiple irreversible oxidation peaks, which have been tentatively attributed to the loss of alkoxide ligand coupled with the deposition of a solid Fe-containing coating on the electrode. This coating was stable out to the voltage limits for the acetonitrile solvent. PMID- 21635012 TI - Fano resonances in nanoscale plasmonic systems: a parameter-free modeling approach. AB - The interaction between plasmonic resonances, sharp modes, and light in nanoscale plasmonic systems often leads to Fano interference effects. This occurs because the plasmonic excitations are usually spectrally broad and the characteristic narrow asymmetric Fano line-shape results upon interaction with spectrally sharper modes. By considering the plasmonic resonance in the Fano model, as opposed to previous flat continuum approaches, here we show that a simple and exact expression for the line-shape can be found. This allows the role of the width and energy of the plasmonic resonance to be properly understood. As examples, we show how Fano resonances measured on an array of gold nanoantennas covered with PMMA, as well as the hybridization of dark with bright plasmons in nanocavities, are well reproduced with a simple exact formula and without any fitting parameters. PMID- 21635013 TI - Palladium(II) complexes of readily functionalized bidentate 2-pyridyl-1,2,3 triazole "click" ligands: a synthetic, structural, spectroscopic, and computational study. AB - The Cu(I)-catalyzed 1,3-cycloaddition of organic azides with terminal alkynes, the CuAAC "click" reaction is currently receiving considerable attention as a mild, modular method for the generation of functionalized ligand scaffolds. Herein we show that mild one-pot "click" methods can be used to readily and rapidly synthesize a family of functionalized bidentate 2-pyridyl-1,2,3-triazole ligands, containing electrochemically, photochemically, and biologically active functional groups in good to excellent yields (47-94%). The new ligands have been fully characterized by elemental analysis, HR-ESI-MS, IR, (1)H and (13)C NMR and in three cases by X-ray crystallography. Furthermore we have demonstrated that this family of functionalized "click" ligands readily form bis-bidentate Pd(II) complexes. Solution studies, X-ray crystallography, and density functional theory (DFT) calculations indicate that the Pd(II) complexes formed with the 2-(1-R-1H 1,2,3-triazol-4-yl)pyridine series of ligands are more stable than those formed with the [4-R-1H-1,2,3-triazol-1-yl)methyl]pyridine "click" ligands. PMID- 21635014 TI - Nonlinear mixing in nanowire subwavelength waveguides. AB - The realization of nonlinear photonic circuits to achieve the control of light-by light is contingent upon a strong nonlinear response that can be captured in a guided-wave geometry. There remains a need to further scale down waveguides while maintaining a strong nonlinear response. In this study, we report second-harmonic generation and optical parametric generation using the second-order nonlinear response in an 80 nm thick CdS nanowire subwavelength waveguide. Moreover, our three-dimensional finite-difference time-domain (FDTD) simulations demonstrate that it is possible to enhance the coherence length due to the very nature of the subwavelength geometry. Nonlinear mixing in a nanowire subwavelength waveguide represents an advance toward all-optical processing and all-optical switching in integrated photonic circuits. PMID- 21635015 TI - Controlled swapping of nanocomposite surface wettability by multilayer photopolymerization. AB - Single-layered photopolymerized nanocomposite films of polystyrene and TiO(2) nanorods change their wetting characteristics from hydrophobic to hydrophilic when deposited on substrates with decreasing hydrophilicity. Interestingly, the addition of a second photopolymerized layer causes a swapping in the wettability, so that the final samples result converted from hydrophobic to hydrophilic or vice versa. The wettability characteristics continue to be swapped as the number of photopolymerized layers increases. In fact, odd-layered samples show the same wetting behavior as single-layered ones, while even-layered samples have the same surface characteristics as double-layered ones. Analytical surface studies demonstrate that all samples, independently of the number of layers, have similar low roughness, and that the wettability swap is due to the different concentration of the nanocomposites constituents on the samples surface. Particularly, the different interactions between the hydrophilic TiO(2) nanorods and the underlying layer lead to different amounts of nanorods exposed on the nanocomposites surface. Moreover, due to the unique property of TiO(2) to reversibly increase its wettability upon UV irradiation and subsequent storage, the wetting characteristics of the multilayered nanocomposites can be tuned in a reversible manner. In this way, a combination of substrate, number of photopolymerized layers, and external UV light stimulus can be used in order to precisely control the surface wettability properties of nanocomposite films, opening the way to a vast number of potential applications in microfluidics, protein assays, and cell growth. PMID- 21635016 TI - Complete family of mono-, bi-, and trinuclear Re(I)(CO)3Cl complexes of the bridging polypyridyl ligand 2,3,8,9,14,15-hexamethyl-5,6,11,12,17,18 hexaazatrinapthalene: syn/anti isomer separation, characterization, and photophysics. AB - The syn and anti isomers of the bi- and trinuclear Re(CO)(3)Cl complexes of 2,3,8,9,14,15-hexamethyl-5,6,11,12,17,18-hexaazatrinapthalene (HATN-Me(6)) are reported. The isomers are characterized by (1)H NMR spectroscopy and X-ray crystallography. The formation of the binuclear complex from the reaction of HATN Me(6) with 2 equiv of Re(CO)(5)Cl in chloroform results in a 1:1 ratio of the syn and anti isomers. However, synthesis of the trinuclear complex from the reaction of HATN-Me(6) with 3 equiv of Re(CO)(5)Cl in chloroform produces only the anti isomer. syn-{(Re(CO)(3)Cl)(3)(MU-HATN-Me(6))} can be synthesized by reacting 1 equiv of Re(CO)(5)Cl with syn-{(Re(CO)(3)Cl)(2)(MU-HATN-Me(6))} in refluxing toluene. The product is isolated by subsequent chromatography. The X-ray crystal structures of syn-{(Re(CO)(3)Cl)(2)(MU-HATN-Me(6))} and anti-{(Re(CO)(3)Cl)(3)(MU HATN-Me(6))} are presented both showing severe distortions of the HATN ligand unit and intermolecular pi stacking. The complexes show intense absorptions in the visible region, comprising strong pi -> pi* and metal-to-ligand charge transfer (MLCT) transitions, which are modeled using time-dependent density functional theory (TD-DFT). The energy of the MLCT absorption decreases from mono to bi- to trinuclear complexes. The first reduction potentials of the complexes become more positive upon binding of subsequent Re(CO)(3)Cl fragments, consistent with changes in the energy of the MLCT bands and lowering of the energy of relevant lowest unoccupied molecular orbitals, and this is supported by TD-DFT. The nature of the excited states of all of the complexes is also studied using both resonance Raman and picosecond time-resolved IR spectroscopy, where it is shown that MLCT excitation results in the oxidation of one rhenium center. The patterns of the shifts in the carbonyl bands upon excitation reveal that the MLCT state is localized on one rhenium center on the IR time scale. PMID- 21635017 TI - Estimating the relationship between preference-based generic utility instruments and disease-specific quality-of-life measures in severe chronic constipation: challenges in practice. AB - BACKGROUND: Many clinical trials that generate evidence on the quality-of-life (QOL) improvements provided by new health technologies do not incorporate a preference-based generic measure, but generate only disease-specific data. However, in order to meet the information needs of regulators such as the UK National Institute for Health and Clinical Excellence (NICE), such disease specific data need to be converted into a broader generic measure; for NICE, the preferred instrument is the EQ-5D. The process of converting QOL data from one instrument to another is known as 'mapping'. OBJECTIVE: The objective of this study was to examine the extent to which disease-specific measures generated in the clinical trials for a new treatment for chronic constipation (prucalopride) can be 'mapped' onto a preference-based generic measure (EQ-5D and SF-6D) to generate robust and reliable utility estimates. METHODS: Disease-specific QOL data generated in the clinical trials of prucalopride (PAC-QOL scores) were converted into utility values estimated using the preference-based generic measure EQ-5D. SF-36 data were also collected in the clinical trials and used to generate SF-6D estimates for comparative purposes. Regression analysis was used to derive a range of mapping functions to identify the extent to which increasing the complexity of the hypothesized underlying mapping function enhanced the robustness and reliability of the obtained mapping relationship. RESULTS: The mean utility observed at baseline for chronic constipation, based on SF-36 data, was 0.813 with the EQ-5D and 0.723 with the SF-6D. An examination of the differences between predicted and observed values generally found that the mapping functions generated were robust and reliable, with little evidence of bias across the range of the dependent variable. However, the nature of the symptoms explored in the PAC-QOL measure was, in general, less severe than those explored in the EQ-5D. For example, the condition-specific measures explored the degree to which patients experienced 'discomfort', rather than 'pain' as evaluated in the EQ-5D. Given this limitation in the severity range covered in the disease-specific measures, it is perhaps not surprising that a 'floor effect' was identified, with certain health dimensions mapping only to the upper range of the EQ-5D measure. CONCLUSIONS: In circumstances where direct utility measurement is not available, mapping provides a valuable method by which to estimate utility data for incorporation into cost-effectiveness analyses. Our findings emphasize the importance of the structure and nature of the mapping analysis undertaken as being a fundamental determinant of the utility estimates generated. Unfortunately, the theoretical guidance available to steer such analyses is still comparatively underdeveloped and this remains an area of health economic analysis in which empiricism largely rules. Ensuring that such mapping is undertaken and interpreted in as transparent and robust a manner as possible is therefore crucial in allowing regulators to accurately compare the clinical and cost effectiveness of new drugs across therapeutic areas. PMID- 21635018 TI - Rydberg-induced solitons: three-dimensional self-trapping of matter waves. AB - We propose a scheme for the creation of stable three-dimensional bright solitons in Bose-Einstein condensates, i.e., the matter-wave analog of so-called spatiotemporal "light bullets." Off-resonant dressing to Rydberg nD states is shown to provide nonlocal attractive interactions, leading to self-trapping of mesoscopic atomic clouds by a collective excitation of a Rydberg atom pair. We present detailed potential calculations and demonstrate the existence of stable solitons under realistic experimental conditions by means of numerical simulations. PMID- 21635019 TI - Temperature dependence of the universal contact parameter in a unitary Fermi gas. AB - The contact I, introduced by Tan, has emerged as a key parameter characterizing universal properties of strongly interacting Fermi gases. For ultracold Fermi gases near a Feshbach resonance, the contact depends upon two quantities: the interaction parameter 1/(k(F)a), where k(F) is the Fermi wave vector and a is the s-wave scattering length, and the temperature T/T(F), where T(F) is the Fermi temperature. We present the first measurements of the temperature dependence of the contact in a unitary Fermi gas using Bragg spectroscopy. The contact is seen to follow the predicted decay with temperature and shows how pair-correlations at high momentum persist well above the superfluid transition temperature. PMID- 21635020 TI - Fractionalization via Z(2) gauge fields at a cold-atom quantum Hall transition. AB - We study a single species of fermionic atoms in an "effective" magnetic field at total filling factor nu(f)=1, interacting through a p-wave Feshbach resonance, and show that the system undergoes a quantum phase transition from a nu(f)=1 fermionic integer quantum Hall state to nu(b)=1/4 bosonic fractional quantum Hall state as a function of detuning. The transition is in the (2+1)D Ising universality class. We formulate a dual theory in terms of quasiparticles interacting with a Z(2) gauge field and show that charge fractionalization follows from this topological quantum phase transition. Experimental consequences and possible tests of our theoretical predictions are discussed. PMID- 21635021 TI - Tuned transition from quantum to classical for macroscopic quantum states. AB - The boundary between the classical and quantum worlds has been intensely studied. It remains fascinating to explore how far the quantum concept can reach with use of specially fabricated elements. Here we employ a tunable flux qubit with basis states having persistent currents of 1 MUA carried by a million pairs of electrons. By tuning the tunnel barrier between these states we see a crossover from quantum to classical. Released from nonequilibrium, the system exhibits spontaneous coherent oscillations. For high barriers the lifetime of the states increases dramatically while the tunneling period approaches the phase coherence time and the oscillations fade away. PMID- 21635022 TI - Complete reconstruction of the wave function of a reacting molecule by four-wave mixing spectroscopy. AB - Probing the real time dynamics of a reacting molecule remains one of the central challenges in chemistry. Here we show how the time-dependent wave function of an excited-state reacting molecule can be completely reconstructed from resonant coherent anti-Stokes Raman spectroscopy. The method assumes knowledge of the ground potential but not of any excited potential. The excited-state potential can in turn be constructed from the wave function. The formulation is general for polyatomics and applies to bound as well as dissociative excited potentials. We demonstrate the method on the Li(2) molecule. PMID- 21635023 TI - Quantum simulation of time-dependent Hamiltonians and the convenient illusion of Hilbert space. AB - We consider the manifold of all quantum many-body states that can be generated by arbitrary time-dependent local Hamiltonians in a time that scales polynomially in the system size, and show that it occupies an exponentially small volume in Hilbert space. This implies that the overwhelming majority of states in Hilbert space are not physical as they can only be produced after an exponentially long time. We establish this fact by making use of a time-dependent generalization of the Suzuki-Trotter expansion, followed by a well-known counting argument. This also demonstrates that a computational model based on arbitrarily rapidly changing Hamiltonians is no more powerful than the standard quantum circuit model. PMID- 21635024 TI - Extension of loop quantum gravity to f(R) theories. AB - The four-dimensional metric f(R) theories of gravity are cast into connection dynamical formalism with real su(2) connections as configuration variables. Through this formalism, the classical metric f(R) theories are quantized by extending the loop quantization scheme of general relativity. Our results imply that the nonperturbative quantization procedure of loop quantum gravity is valid not only for general relativity but also for a rather general class of four dimensional metric theories of gravity. PMID- 21635025 TI - Cores in dwarf galaxies from dark matter with a Yukawa potential. AB - We show that cold dark matter particles interacting through a Yukawa potential could naturally explain the recently observed cores in dwarf galaxies without affecting the dynamics of objects with a much larger velocity dispersion, such as clusters of galaxies. The velocity dependence of the associated cross section as well as the possible exothermic nature of the interaction alleviates earlier concerns about strongly interacting dark matter. Dark matter evaporation in low mass objects might explain the observed deficit of satellite galaxies in the Milky Way halo and have important implications for the first galaxies and reionization. PMID- 21635026 TI - New massive gravity and AdS(4) counterterms. AB - We show that the recently proposed Dirac-Born-Infeld extension of new massive gravity emerges naturally as a counterterm in four-dimensional anti-de Sitter space (AdS(4)). The resulting on-shell Euclidean action is independent of the cutoff at zero temperature. We also find that the same choice of counterterm gives the usual area law for the AdS(4) Schwarzschild black hole entropy in a cutoff-independent manner. The parameter values of the resulting counterterm action correspond to a c=0 theory in the context of the duality between AdS(3) gravity and two-dimensional conformal field theory. We rewrite this theory in terms of the gauge field that is used to recast 3D gravity as a Chern-Simons theory. PMID- 21635029 TI - Dihadron fragmentation functions for large invariant mass. AB - Using perturbative quantum chromodynamics, we compute dihadron fragmentation functions for a large invariant mass of the dihadron pair. The main focus is on the interference fragmentation function H(1)(?), which plays an important role in spin physics of the nucleon. Our calculation also reveals that H(1)(?) and the Collins fragmentation function have closely related underlying dynamics. By considering semi-inclusive deep-inelastic scattering, we further show that collinear factorization in terms of dihadron fragmentation functions and collinear factorization in terms of single-hadron fragmentation functions provide the same result in the region of intermediate invariant mass. PMID- 21635027 TI - Invariant mass distribution of jet pairs produced in association with a W boson in pp collisions at sqrt[s]=1.96 TeV. AB - We report a study of the invariant mass distribution of jet pairs produced in association with a W boson using data collected with the CDF detector which correspond to an integrated luminosity of 4.3 fb(-1). The observed distribution has an excess in the 120-160 GeV/c(2) mass range which is not described by current theoretical predictions within the statistical and systematic uncertainties. In this Letter, we report studies of the properties of this excess. PMID- 21635028 TI - Search for the standard model Higgs boson in the H->WW->lnuq'q decay channel. AB - We present a search for the standard model Higgs boson (H) in pp collisions at sqrt(s)=1.96 TeV in events containing a charged lepton (l), missing transverse energy, and at least two jets, using 5.4 fb(-1) of integrated luminosity recorded with the D0 detector at the Fermilab Tevatron Collider. This analysis is sensitive primarily to Higgs bosons produced through the fusion of two gluons or two electroweak bosons, with subsequent decay H->WW->lnuq'q, where l is an electron or muon. The search is also sensitive to contributions from other production channels, such as WH->lnubb. In the absence of a signal, we set limits at the 95% C.L. on the cross section for H production sigma(pp->H+X) in these final states. For a mass of M(H)=160 GeV, the limit is a factor of 3.9 larger than the cross section in the standard model and consistent with an a priori expected sensitivity of 5.0. PMID- 21635033 TI - Tunnel ionization of molecules and orbital imaging. AB - We study whether tunnel ionization of aligned molecules can be used to map out the electronic structure of the ionizing orbitals. We show that the common view, which associates tunnel ionization rates with the electronic density profile of the ionizing orbital, is not always correct. Using the example of tunnel ionization from the CO(2) molecule, we show how and why the angular structure of the alignment-dependent ionization rate moves with increasing the strength of the electric field. These modifications reflect a general trend for molecules. PMID- 21635031 TI - Phase diagram and critical end point for strongly interacting quarks. AB - We introduce a method based on chiral susceptibility, which enables one to draw a phase diagram in the chemical-potential-temperature plane for strongly interacting quarks whose interactions are described by any reasonable gap equation, even if the diagrammatic content of the quark-gluon vertex is unknown. We locate a critical end point at (MU(E),T(E))~(1.0,0.9)T(c), where T(c) is the critical temperature for chiral-symmetry restoration at MU=0, and find that a domain of phase coexistence opens at the critical end point whose area increases as a confinement length scale grows. PMID- 21635032 TI - Fusion reactions with the one-neutron halo nucleus (15)C. AB - The structure of (15)C, with an s(1/2) neutron weakly bound to a closed-neutron shell nucleus (14)C, makes it a prime candidate for a one-neutron halo nucleus. We have for the first time studied the cross section for the fusion-fission reaction (15)C+(232)Th at energies in the vicinity of the Coulomb barrier and compared it to the yield of the neighboring (14)C+(232)Th system measured in the same experiment. At sub-barrier energies, an enhancement of the fusion yield by factors of 2-5 was observed for (15)C, while the cross sections for (14)C match the trends measured for (12,13)C. PMID- 21635034 TI - Proposal for a laser control of vibrational cooling in Na(2) using resonance coalescence. AB - With a specific choice of laser parameters resulting in a so-called exceptional point (EP) in the wavelength-intensity parameter plane, it is possible to produce the coalescence of two Floquet resonances describing the photodissociation of the Na(2) molecule, which is one of the candidates for the formation of samples of translationally cold molecules. By appropriately tuning laser parameters along a contour encircling the exceptional point, the resonances exchange their quantum nature. Thus a laser-controlled transfer of the probability density from one field-free vibrational level to another is achieved through adiabatic transport involving these resonances. We propose an efficient scenario for vibrational cooling of Na(2) referring to cascade transfers involving multiple EPs and predicted to be robust up to a 78% rate against laser-induced dissociation. PMID- 21635035 TI - Laser excitation of positronium in the Paschen-Back regime. AB - Zeeman mixing of singlet and triplet 2P states of positronium (Ps) atoms, followed by decay back to the ground state, can effectively turn a long-lived triplet atom into a short-lived singlet state, which would seem to preclude laser cooling of Ps in a magnetic field. Here we report experiments which show that, in fact, because of the large splitting of the n=2 states in a high magnetic field (the Paschen-Back regime), the amount of such mixing diminishes approximately exponentially with an increasing magnetic field >0.01 T and is essentially eliminated above ~2 T. Thus, laser cooling of Ps should be feasible at high fields, which will facilitate the production of a Ps Bose-Einstein condensate. PMID- 21635036 TI - Wrapping an adhesive sphere with an elastic sheet. AB - We study the adhesion of an elastic sheet on a rigid spherical substrate. Gauss's Theorema Egregium shows that this operation necessarily generates metric distortions (i.e., stretching) as well as bending. As a result, a large variety of contact patterns ranging from simple disks to complex branched shapes are observed as a function of both geometrical and material properties. We describe these different morphologies as a function of two nondimensional parameters comparing, respectively, bending and stretching energies to adhesion. A complete configuration diagram is finally proposed. PMID- 21635037 TI - Negative effective gravity in water waves by periodic resonator arrays. AB - Based on analytic derivations and numerical simulations, we show that near a low resonant frequency water waves cannot propagate through a periodic array of resonators (bottom-mounted split tubes) as if water has a negative effective gravitational acceleration g(e) and positive effective depth h(e). This gives rise to a low-frequency resonant band gap in which water waves can be strongly reflected by the resonator array. For a damping resonator array, the resonant gap can also dramatically modify the absorption efficiency of water waves. The results provide a mechanism to block water waves and should find applications in ocean wave energy extraction. PMID- 21635038 TI - Compressible turbulence: the cascade and its locality. AB - We prove that interscale transfer of kinetic energy in compressible turbulence is dominated by local interactions. In particular, our results preclude direct transfer of kinetic energy from large-scales to dissipation scales, such as into shocks, in high Reynolds number turbulence as is commonly believed. Our assumptions on the scaling of structure functions are weak and enjoy compelling empirical support. Under a stronger assumption on pressure dilatation cospectrum, we show that mean kinetic and internal energy budgets statistically decouple beyond a transitional conversion range. Our analysis establishes the existence of an ensuing inertial range over which mean subgrid scale kinetic energy flux becomes constant, independent of scale. Over this inertial range, mean kinetic energy cascades locally and in a conservative fashion despite not being an invariant. PMID- 21635039 TI - Plastic deformations in complex plasmas. AB - Complex plasmas are macroscopic model systems of real solids and liquids, used to study underdamped dynamics and wave phenomena. Plastic deformations of complex plasma crystals under slow uniaxial compression have been studied experimentally and numerically. It is shown that the lattice becomes locally sheared and that this strain is relaxed by shear slips resulting in global uniform compression and heat generation. Shear slips generate pairs of dislocations which move in opposite directions at subsonic speeds. PMID- 21635040 TI - Time evolution of collisionless shock in counterstreaming laser-produced plasmas. AB - We investigated the time evolution of a strong collisionless shock in counterstreaming plasmas produced using a high-power laser pulse. The counterstreaming plasmas were generated by irradiating a CH double-plane target with the laser. In self-emission streaked optical pyrometry data, steepening of the self-emission profile as the two-plasma interaction evolved indicated shock formation. The shock thickness was less than the mean free path of the counterstreaming ions. Two-dimensional snapshots of the self-emission and shadowgrams also showed very thin shock structures. The Mach numbers estimated from the flow velocity and the brightness temperatures are very high. PMID- 21635041 TI - High magnetic shear gain in a liquid sodium stable Couette flow experiment: a prelude to an alpha-Omega dynamo. AB - The Omega phase of the liquid sodium alpha-Omega dynamo experiment at New Mexico Institute of Mining and Technology in cooperation with Los Alamos National Laboratory has demonstrated a high toroidal field B(phi) that is ?8*B(r), where B(r) is the radial component of an applied poloidal magnetic field. This enhanced toroidal field is produced by the rotational shear in stable Couette flow within liquid sodium at a magnetic Reynolds number Rm?120. Small turbulence in stable Taylor-Couette flow is caused by Ekman flow at the end walls, which causes an estimated turbulence energy fraction of (deltav/v)(2)~10(-3). PMID- 21635042 TI - Turbulent transport in tokamak plasmas with rotational shear. AB - Nonlinear gyrokinetic simulations are conducted to investigate turbulent transport in tokamak plasmas with rotational shear. At sufficiently large flow shears, linear instabilities are suppressed, but transiently growing modes drive subcritical turbulence whose amplitude increases with flow shear. This leads to a local minimum in the heat flux, indicating an optimal E*B shear value for plasma confinement. Local maxima in the momentum fluxes are observed, implying the possibility of bifurcations in the E*B shear. The critical temperature gradient for the onset of turbulence increases with flow shear at low flow shears; at higher flow shears, the dependence of heat flux on temperature gradient becomes less stiff. The turbulent Prandtl number is found to be largely independent of temperature and flow gradients, with a value close to unity. PMID- 21635043 TI - Optical flux lattices for ultracold atomic gases. AB - We show that simple laser configurations can give rise to "optical flux lattices," in which optically dressed atoms experience a periodic effective magnetic flux with high mean density. These potentials lead to narrow energy bands with nonzero Chern numbers. Optical flux lattices will greatly facilitate the achievement of the quantum Hall regime for ultracold atomic gases. PMID- 21635044 TI - Experimental verification of the formation mechanism for pillar arrays in nanofilms subject to large thermal gradients. AB - The free surface of molten nanofilms is known to undergo spontaneous formation of periodic protrusions when exposed to a large transverse thermal gradient. Early time measurements of the array pitch and growth rate in polymer melts confirm a formation process based on a long wavelength thermocapillary instability and not electrostatic attraction or acoustic phonon driven growth as previously believed. We find excellent agreement with theoretical predictions provided the nanofilm out-of-plane thermal conductivity is several times larger than bulk, an enhancement suggestive of polymer chain alignment. PMID- 21635045 TI - Theory, synthesis, and oxygen reduction catalysis of Fe-porphyrin-like carbon nanotube. AB - We report the synthesis of a Fe-porphyrin-like carbon nanotube from conventional plasma-enhanced chemical vapor deposition. Covalent but seamless incorporation of the 5-6-5-6 porphyrinic Fe-N(4) moiety into the graphene hexagonal side wall was elucidated by x-ray and ultraviolet photoemission spectroscopies and first principles electronic structure calculations. The resulting biomimetic nanotube exhibits an excellent oxygen reduction catalytic activity with the extreme structural stability over 0.1*10(6) cycles, vastly superior to the commercial Pt C catalyst. PMID- 21635046 TI - Minimal integer automaton behind crystal plasticity. AB - Power law fluctuations and scale-free spatial patterns are known to characterize steady state plastic flow in crystalline materials. In this Letter we study the emergence of correlations in a simple Frenkel-Kontorova-type model of 2D plasticity which is largely free of arbitrariness, amenable to analytical study, and is capable of generating critical exponents matching experiments. Our main observation concerns the possibility to reduce continuum plasticity to an integer valued automaton revealing inherent discreteness of the plastic flow. PMID- 21635047 TI - High resolution transmission electron microscope observation of zero-strain deformation twinning mechanisms in Ag. AB - We have observed a new deformation-twinning mechanism using the high resolution transmission electron microscope in polycrystalline Ag films, zero-strain twinning via nucleation, and the migration of a Sigma3{112} incoherent twin boundary (ITB). This twinning mechanism produces a near zero macroscopic strain because the net Burgers vectors either equal zero or are equivalent to a Shockley partial dislocation. This observation provides new insight into the understanding of deformation twinning and confirms a previous hypothesis: detwinning could be accomplished via the nucleation and migration of Sigma3{112} ITBs. The zero strain twinning mechanism may be unique to low staking fault energy metals with implications for their deformation behavior. PMID- 21635048 TI - Collective hypersonic excitations in strongly multiple scattering colloids. AB - Unprecedented low-dispersion high-frequency acoustic excitations are observed in dense suspensions of elastically hard colloids. The experimental phononic band structure for SiO(2) particles with different sizes and volume fractions is well represented by rigorous full-elastodynamic multiple-scattering calculations. The slow phonons, which do not relate to particle resonances, are localized in the surrounding liquid medium and stem from coherent multiple scattering that becomes strong in the close-packing regime. Such rich phonon-matter interactions in nanostructures, being still unexplored, can open new opportunities in phononics. PMID- 21635049 TI - Numerical evidence for critical behavior of the two-dimensional nonequilibrium zero-temperature random field Ising model. AB - We give numerical evidence that the two-dimensional nonequilibrium zero temperature random field Ising model exhibits critical behavior. Our findings are based on the results of scaling analysis and collapsing of data, obtained in extensive simulations of systems with sizes sufficiently large to clearly display the critical behavior. PMID- 21635050 TI - Modeling multiple time scales during glass formation with phase-field crystals. AB - The dynamics of glass formation in monatomic and binary liquids are studied numerically using a microscopic field theory for the evolution of the time averaged atomic number density. A stochastic framework combining phase-field crystal free energies and dynamic density functional theory is shown to successfully describe several aspects of glass formation over multiple time scales. Agreement with mode coupling theory is demonstrated for underdamped liquids at moderate supercoolings, and a rapidly growing dynamic correlation length is found to be associated with fragile behavior. PMID- 21635051 TI - Forces and currents in carbon nanostructures: are we imaging atoms? AB - First-principles calculations show that the rich variety of image patterns found in carbon nanostructures with the atomic force and scanning tunneling microscopes can be rationalized in terms of the chemical reactivity of the tip and the distance range explored in the experiments. For weakly reactive tips, the Pauli repulsion dominates the atomic contrast and force maxima are expected on low electronic density positions as the hollow site. With reactive tips, the interaction is strong enough to change locally the hybridization of the carbon atoms, making it possible to observe atomic resolution in both the attractive and the repulsive regime although with inverted contrast. Regarding STM images, we show that in the near-contact regime, due to current saturation, bright spots correspond to hollow positions instead of atomic sites, providing an explanation for the most common hexagonal pattern found in the experiments. PMID- 21635052 TI - Vacant-site octahedral tilings on SrTiO(3) (001), the (sqrt[13]*sqrt[13])R33.7 degrees surface, and related structures. AB - The structure of the SrTiO(3) (001) (sqrt[13]*sqrt[13])R33.7 degrees surface reconstruction has been determined using transmission electron diffraction combined with direct methods and density functional theory. It has a TiO(2)-rich surface with a 2D tiling of edge or corner-sharing TiO(5)? octahedra. Additionally, different arrangements of these octahedral units at the surface, dictated by local bond-valence sums, form 2D networks that can account for many ordered surface reconstructions as well as disordered glasslike structures consistent with the multitude of structures observed experimentally, and potentially other materials and interfaces. PMID- 21635053 TI - Polariton condensation in a one-dimensional disordered potential. AB - We study the coherence and density modulation of a nonequilibrium exciton polariton condensate in a one-dimensional valley with disorder. By means of interferometric measurements we evidence a modulation of the first-order coherence function and we relate it to a disorder-induced modulation of the condensate density, that increases as the pump power is increased. The nonmonotonic spatial coherence function is found to be the result of the strong nonequilibrium character of the one-dimensional system, in the presence of disorder. PMID- 21635054 TI - Evolution of the Kondo effect in a quantum dot probed by shot noise. AB - We measure the current and shot noise in a quantum dot in the Kondo regime to address the nonequilibrium properties of the Kondo effect. By systematically tuning the temperature and gate voltages to define the level positions in the quantum dot, we observe an enhancement of the shot noise as temperature decreases below the Kondo temperature, which indicates that the two-particle scattering process grows as the Kondo state evolves. Below the Kondo temperature, the Fano factor defined at finite temperature is found to exceed the expected value of unity from the noninteracting model, reaching 1.8+/-0.2. PMID- 21635055 TI - Physical fault tolerance of nanoelectronics. AB - The error rate in complementary transistor circuits is suppressed exponentially in electron number, arising from an intrinsic physical implementation of fault tolerant error correction. Contrariwise, explicit assembly of gates into the most efficient known fault-tolerant architecture is characterized by a subexponential suppression of error rate with electron number, and incurs significant overhead in wiring and complexity. We conclude that it is more efficient to prevent logical errors with physical fault tolerance than to correct logical errors with fault-tolerant architecture. PMID- 21635056 TI - Valley-dependent Brewster angles and Goos-Hanchen effect in strained graphene. AB - We demonstrate theoretically how local strains in graphene can be tailored to generate a valley-polarized current. By suitable engineering of local strain profiles, we find that electrons in opposite valleys (K or K') show different Brewster-like angles and Goos-Hanchen shifts, exhibiting a close analogy with light propagating behavior. In a strain-induced waveguide, electrons in K and K' valleys have different group velocities, which can be used to construct a valley filter in graphene without the need for any external fields. PMID- 21635057 TI - Antiferroquadrupolar ordering in a Pr-based superconductor PrIr(2)Zn(20). AB - An antiferroquadrupolar ordering at T(Q)=0.11 K has been found in a Pr-based superconductor PrIr(2)Zn(20). The measurements of specific heat and magnetization revealed the non-Kramers Gamma(3) doublet ground state with the quadrupolar degrees of freedom. The specific heat exhibits a sharp peak at T(Q)=0.11 K. The increment of T(Q) in magnetic fields and the anisotropic B-T phase diagram are consistent with the antiferroquadrupolar ordered state below T(Q). The entropy release at T(Q) is only 20% of Rln2, suggesting that the quadrupolar fluctuations play a role in the formation of the superconducting pairs below T(c)=0.05 K. PMID- 21635058 TI - Magnetism in closed-shell quantum dots: emergence of magnetic bipolarons. AB - Similar to atoms and nuclei, semiconductor quantum dots exhibit the formation of shells. Predictions of magnetic behavior of the dots are often based on the shell occupancies. Thus, closed-shell quantum dots are assumed to be inherently nonmagnetic. Here, we propose a possibility of magnetism in such dots doped with magnetic impurities. On the example of the system of two interacting fermions, the simplest embodiment of the closed-shell structure, we demonstrate the emergence of a novel broken-symmetry ground state that is neither spin singlet nor spin triplet. We propose experimental tests of our predictions and the magnetic-dot structures to perform them. PMID- 21635059 TI - Low-temperature dynamics of magnons in a spin-1/2 ladder compound. AB - We have used a combination of neutron resonant spin-echo and triple-axis spectroscopies to determine the energy, fine structure, and linewidth of the magnon resonance in the model spin-1/2 ladder antiferromagnet IPA-CuCl(3) at temperatures T?Delta(0)/k(B), where Delta(0) is the spin gap at T=0. In this low temperature regime we find that the results deviate substantially from the predictions of the nonlinear sigma model proposed as a description of magnon excitations in one-dimensional quantum magnets and attribute these deviations to real-space and spin-space anisotropies in the spin Hamiltonian as well as scattering of magnon excitations from a dilute density of impurities. These effects are generic to experimental realizations of one-dimensional quantum magnets. PMID- 21635060 TI - Magnetic couplings in CsV(2)O(5): a new picture. AB - Concerning its magnetic properties, the layered vanadate CsV(2)O(5) has long been considered as formed by isolated spin-1/2 dimers characterized by a large antiferromagnetic coupling of about 146 K. This interpretation was supported by both magnetic susceptibility measurements and the obvious presence of magnetically active strongly dimerized V(4+) ions. In this work we investigate the magnetic properties of this compound through an extensive use of the broken symmetry formalism in the framework of density-functional theory. Our calculations demonstrate that the system is built from strongly dimerized alternating chains where the structural and magnetic dimers are distinct from each other. PMID- 21635061 TI - Density-matrix approach for the electroluminescence of molecules in a scanning tunneling microscope. AB - The electroluminescence (EL) of molecules confined inside a nanocavity in the scanning tunneling microscope possesses many intriguing but unexplained features. We present here a general theoretical approach based on the density-matrix formalism to describe the EL from molecules near a metal surface induced by both electron tunneling and localized surface plasmon excitations simultaneously. It reveals the underlying physical mechanism for the external bias dependent EL. The important role played by the localized surface plasmon on the EL is highlighted. Calculations for porphyrin derivatives have reproduced corresponding experimental spectra and nicely explained the observed unusual large variation of emission spectral profiles. This general theoretical approach can find many applications in the design of molecular electronic and photonic devices. PMID- 21635062 TI - Nematic braids: topological invariants and rewiring of disclinations. AB - The conventional topological description given by the fundamental group of nematic order parameter does not adequately explain the entangled defect line structures that have been observed in nematic colloids. We introduce a new topological invariant, the self-linking number, that enables a complete classification of entangled defect line structures in general nematics, even without particles, and demonstrate our formalism using colloidal dimers, for which entangled structures have been previously observed. We also unveil a simple rewiring scheme for the orthogonal crossing of two -1/2 disclinations, based on a tetrahedral rotation of two relevant disclination segments, that allows us to predict possible nematic braids and calculate their self-linking numbers. PMID- 21635063 TI - Hyperuniform long-range correlations are a signature of disordered jammed hard particle packings. AB - We show that quasi-long-range (QLR) pair correlations that decay asymptotically with scaling r(-(d+1)) in d-dimensional Euclidean space R(d), trademarks of certain quantum systems and cosmological structures, are a universal signature of maximally random jammed (MRJ) hard-particle packings. We introduce a novel hyperuniformity descriptor in MRJ packings by studying local-volume-fraction fluctuations and show that infinite-wavelength fluctuations vanish even for packings with size and shape distributions. Special void statistics induce hyperuniformity and QLR pair correlations. PMID- 21635064 TI - Enhanced diffusion by reciprocal swimming. AB - Purcell's scallop theorem states that swimmers deforming their shapes in a time reversible manner ("reciprocal" motion) cannot swim. Using numerical simulations and theoretical calculations we show here that, in a fluctuating environment, reciprocal swimmers undergo, on time scales larger than that of their rotational diffusion, diffusive dynamics with enhanced diffusivities, possibly by orders of magnitude, above normal translational diffusion. Reciprocal actuation does therefore lead to a significant advantage over nonmotile behavior for small organisms such as marine bacteria. PMID- 21635065 TI - Nonlinear pressure dependence of the interaction potential of dense protein solutions. AB - The influence of pressure on the structure and protein-protein interaction potential of dense protein solutions was studied and analyzed using small-angle x ray scattering in combination with a liquid state theoretical approach. The structural as well as the interaction parameters of dense lysozyme solutions are affected by pressure in a nonlinear way. The structural properties of water lead to a modification of the protein-protein interactions below 4 kbar, which might have significant consequences for the stability of proteins in extreme natural environments. PMID- 21635066 TI - Paddling mode of forward flight in insects. AB - By analyzing high-speed video of the fruit fly, we discover a swimminglike mode of forward flight characterized by paddling wing motions. We develop a new aerodynamic analysis procedure to show that these insects generate drag-based thrust by slicing their wings forward at low angle of attack and pushing backwards at a higher angle. Reduced-order models and simulations reveal that the law for flight speed is determined by these wing motions but is insensitive to material properties of the fluid. Thus, paddling is as effective in air as in water and represents a common strategy for propulsion through aquatic and aerial environments. PMID- 21635067 TI - Optimal branching asymmetry of hydrodynamic pulsatile trees. AB - Most of the studies on optimal transport are done for steady state regime conditions. Yet, there exists numerous examples in living systems where supply tree networks have to deliver products in a limited time due to the pulsatile character of the flow, as it is the case for mammalian respiration. We report here that introducing a systematic branching asymmetry allows the tree to reduce the average delivery time of the products. It simultaneously increases its robustness against the inevitable variability of sizes related to morphogenesis. We then apply this approach to the human tracheobronchial tree. We show that in this case all extremities are supplied with fresh air, provided that the asymmetry is smaller than a critical threshold which happens to match the asymmetry measured in the human lung. This could indicate that the structure is tuned at the maximum asymmetry level that allows the lung to feed all terminal units with fresh air. PMID- 21635068 TI - Insulator-to-metal transition in sulfur-doped silicon. AB - We observe an insulator-to-metal transition in crystalline silicon doped with sulfur to nonequilibrium concentrations using ion implantation followed by pulsed laser melting and rapid resolidification. This insulator-to-metal transition is due to a dopant known to produce only deep levels at equilibrium concentrations. Temperature-dependent conductivity and Hall effect measurements for temperatures T>1.7 K both indicate that a transition from insulating to metallic conduction occurs at a sulfur concentration between 1.8 and 4.3*10(20) cm(-3). Conduction in insulating samples is consistent with variable-range hopping with a Coulomb gap. The capacity for deep states to effect metallic conduction by delocalization is the only known route to bulk intermediate band photovoltaics in silicon. PMID- 21635069 TI - Cavity bounds on higher-order lorentz-violating coefficients. AB - We determine the sensitivity of a modern Michelson-Morley resonant-cavity experiment to higher-order nonbirefringent and nondispersive coefficients of the Lorentz-violating standard-model extension. Data from a recent year-long run of the experiment are used to place the first experimental bounds on coefficients associated with nonrenormalizable Lorentz-violating operators. PMID- 21635070 TI - Correlation complementarity yields bell monogamy relations. AB - We present a method to derive Bell monogamy relations by connecting the complementarity principle with quantum nonlocality. The resulting monogamy relations are stronger than those obtained from the no-signaling principle alone. In many cases, they yield tight quantum bounds on the amount of violation of single and multiple qubit correlation Bell inequalities. In contrast with the two qubit case, a rich structure of possible violation patterns is shown to exist in the multipartite scenario. PMID- 21635071 TI - Decoherence and disorder in quantum walks: from ballistic spread to localization. AB - We investigate the impact of decoherence and static disorder on the dynamics of quantum particles moving in a periodic lattice. Our experiment relies on the photonic implementation of a one-dimensional quantum walk. The pure quantum evolution is characterized by a ballistic spread of a photon's wave packet along 28 steps. By applying controlled time-dependent operations we simulate three different environmental influences on the system, resulting in a fast ballistic spread, a diffusive classical walk, and the first Anderson localization in a discrete quantum walk architecture. PMID- 21635072 TI - Experimental demonstration of probabilistic quantum cloning. AB - The method of quantum cloning is divided into two main categories: approximate and probabilistic quantum cloning. The former method is used to approximate an unknown quantum state deterministically, and the latter can be used to faithfully copy the state probabilistically. Thus far, many approximate cloning machines have been experimentally demonstrated, but probabilistic cloning remains an experimental challenge, as it requires more complicated networks and a higher level of precision control. In this work, we design an efficient quantum network with a limited amount of resources and perform the first experimental demonstration of probabilistic quantum cloning in a NMR quantum computer. In our experiment, the optimal cloning efficiency proposed by Duan and Guo [Phys. Rev. Lett. 80, 4999 (1998)] is achieved. PMID- 21635073 TI - Detecting the drift of quantum sources: not the de Finetti theorem. AB - We propose and analyze a method to detect and characterize the drift of a nonstationary quantum source. It generalizes a standard measurement for detecting phase diffusion of laser fields to quantum systems of arbitrary Hilbert space dimension, qubits in particular. We distinguish diffusive and systematic drifts, and examine how quickly one can determine that a source is drifting. We show that for single-photon wave packets our measurement is implemented by the Hong-Ou Mandel effect. PMID- 21635074 TI - Entanglement in the symmetric sector of n qubits. AB - We discuss the entanglement properties of symmetric states of n qubits. The Majorana representation maps a generic such state into a system of n points on a sphere. Entanglement invariants, either under local unitaries (LU) or stochastic local operations and classical communication (SLOCC), can then be addressed in terms of the relative positions of the Majorana points. In the LU case, an overcomplete set of invariants can be built from the inner product of the radial vectors pointing to these points; this is detailed for the well-documented three qubits case. In the SLOCC case, a cross ratio of related Mobius transformations are shown to play a central role, exemplified here for four qubits. Finally, as a side result, we also analyze the manifold of maximally entangled 3 qubit state, both in the symmetric and generic case. PMID- 21635075 TI - Controllable anisotropic exchange coupling between spin qubits in quantum dots. AB - The exchange coupling between quantum dot spin qubits is isotropic, which restricts the types of quantum gates that can be formed. Here, we propose a method for controlling anisotropic interactions between spins arranged in a bus geometry. The symmetry is broken by an external magnetic field, resulting in XXZ type interactions that can efficiently generate maximally entangled Greenberger Horne-Zeilinger states or universal gate sets for exchange-only quantum computing. We exploit the XXZ couplings to propose a qubit scheme, based on double dots. PMID- 21635076 TI - Scalable and robust randomized benchmarking of quantum processes. AB - In this Letter we propose a fully scalable randomized benchmarking protocol for quantum information processors. We prove that the protocol provides an efficient and reliable estimate of the average error-rate for a set operations (gates) under a very general noise model that allows for both time and gate-dependent errors. In particular we obtain a sequence of fitting models for the observable fidelity decay as a function of a (convergent) perturbative expansion of the gate errors about the mean error. We illustrate the protocol through numerical examples. PMID- 21635077 TI - Perturbation spreading in many-particle systems: a random walk approach. AB - The propagation of an initially localized perturbation via an interacting many particle Hamiltonian dynamics is investigated. We argue that the propagation of the perturbation can be captured by the use of a continuous-time random walk where a single particle is traveling through an active, fluctuating medium. Employing two archetype ergodic many-particle systems, namely, (i) a hard-point gas composed of two unequal masses and (ii) a Fermi-Pasta-Ulam chain, we demonstrate that the corresponding perturbation profiles coincide with the diffusion profiles of the single-particle Levy walk approach. The parameters of the random walk can be related through elementary algebraic expressions to the physical parameters of the corresponding test many-body systems. PMID- 21635078 TI - Random walk over basins of attraction to construct ising energy landscapes. AB - An efficient algorithm is developed to construct disconnectivity graphs by a random walk over basins of attraction. This algorithm can detect a large number of local minima, find energy barriers between them, and estimate local thermal averages over each basin of attraction. It is applied to the Sherrington Kirkpatrick (SK) spin glass Hamiltonian where existing methods have difficulties even for a moderate number of spins. Finite-size results are used to make predictions in the thermodynamic limit that match theoretical approximations and recent findings on the free energy landscapes of SK spin glasses. PMID- 21635079 TI - Virtual resonance and frequency difference generation by van der Waals interaction. AB - The ability to explore the interior of materials for the presence of inhomogeneities was recently demonstrated by mode synthesizing atomic force microscopy [L. Tetard, A. Passian, and T. Thundat, Nature Nanotech. 5, 105 (2009).]. Proposing a semiempirical nonlinear force, we show that difference frequency omega_ generation, regarded as the simplest synthesized mode, occurs optimally when the force is tuned to van der Waals form. From a parametric study of the probe-sample excitation, we show that the predicted omega_ oscillation agrees well with experiments. We then introduce the concept of virtual resonance to show that probe oscillations at omega_ can efficiently be enhanced. PMID- 21635080 TI - First constraint on cosmological variation of the proton-to-electron mass ratio from two independent telescopes. AB - A high signal-to-noise spectrum covering the largest number of hydrogen lines (90 H(2) lines and 6 HD lines) in a high-redshift object was analyzed from an observation along the sight line to the bright quasar source J2123-005 with the Ultraviolet and Visual Echelle Spectrograph on the European Southern Observatory Very Large Telescope (Paranal, Chile). This delivers a constraint on a possible variation of the proton-to-electron mass ratio of DeltaMU/MU=(8.5 +/- 3.6(stat) +/- 2.2(syst))*10(-6) at redshift z(abs) = 2.059, which agrees well with a recently published result on the same system observed at the Keck telescope yielding DeltaMU/MU=(5.6 +/- 5.5(stat) +/- 2.9(syst))*10(-6). Both analyses used the same robust absorption line fitting procedures with detailed consideration of systematic errors. PMID- 21635081 TI - Large non-gaussianity in axion inflation. AB - The inflationary paradigm has enjoyed phenomenological success; however, a compelling particle physics realization is still lacking. Axions are among the best-motivated inflaton candidates, since the flatness of their potential is naturally protected by a shift symmetry. We reconsider the cosmological perturbations in axion inflation, consistently accounting for the coupling to gauge fields cPhiFF, which is generically present in these models. This coupling leads to production of gauge quanta, which provide a new source of inflaton fluctuations, deltaPhi. For c>=10(2)M(p)(-1), these dominate over the vacuum fluctuations, and non-Gaussianity exceeds the current observational bound. This regime is typical for concrete realizations that admit a UV completion; hence, large non-Gaussianity is easily obtained in minimal and natural realizations of inflation. PMID- 21635082 TI - Critical gravity in four dimensions. AB - We study four-dimensional gravity theories that are rendered renormalizable by the inclusion of curvature-squared terms to the usual Einstein action with a cosmological constant. By choosing the parameters appropriately, the massive scalar mode can be eliminated and the massive spin-2 mode can become massless. This "critical" theory may be viewed as a four-dimensional analogue of chiral topologically massive gravity, or of critical "new massive gravity" with a cosmological constant, in three dimensions. We find that the on-shell energy for the remaining massless gravitons vanishes. There are also logarithmic spin-2 modes, which have positive energy. The mass and entropy of standard Schwarzschild type black holes vanish. The critical theory might provide a consistent toy model for quantum gravity in four dimensions. PMID- 21635083 TI - Measurement of the neutrino mass splitting and flavor mixing by MINOS. AB - Measurements of neutrino oscillations using the disappearance of muon neutrinos from the Fermilab NuMI neutrino beam as observed by the two MINOS detectors are reported. New analysis methods have been applied to an enlarged data sample from an exposure of 7.25*10(20) protons on target. A fit to neutrino oscillations yields values of |Deltam(2)|=(2.32(-0.08)(+0.12))*10(-3) eV(2) for the atmospheric mass splitting and sin(2)(2theta)>0.90 (90% C.L.) for the mixing angle. Pure neutrino decay and quantum decoherence hypotheses are excluded at 7 and 9 standard deviations, respectively. PMID- 21635085 TI - Engineering nonclassicality in a mechanical system through photon subtraction. AB - Nonclassical states of a mechanical mode at nonzero temperature are achieved in a scheme that combines radiation-pressure coupling to a light field and photon subtraction. The scheme embodies an original and experimentally realistic way to obtain mesoscopic quantumness by putting together two mature technologies for quantum control. The protocol is quasi-insensitive to mechanical damping. PMID- 21635086 TI - Control of lasing in biomimetic structures with short-range order. AB - We demonstrate lasing in photonic amorphous structures that mimic the isotropic nanostructures which produce noniridescent color in nature. Our experimental and numerical studies reveal that lasing becomes most efficient at certain frequencies, due to enhanced optical confinement by short-range order. The optimal lasing frequency can be tuned by adjusting the structure factor. This work shows that lasing in nanostructures may be effectively improved and manipulated by short-range order. PMID- 21635087 TI - Saturation of the all-optical Kerr effect. AB - Saturation of the intensity dependence of the refractive index is directly computed from ionization rates via a Kramers-Kronig transform. The linear intensity dependence and its dispersion are found to be in excellent agreement with complete quantum mechanical orbital computations. Higher-order terms concur with solutions of the time-dependent Schrodinger equation. Expanding the formalism to all orders up to the ionization potential of the atom, we derive a model for saturation of the Kerr effect. This model widely confirms recently published and controversially discussed experimental data and corroborates the importance of higher-order Kerr terms for filamentation. PMID- 21635088 TI - Stimulated emission of surface plasmon polaritons in a microcylinder cavity. AB - We have observed stimulated emission of surface plasmon polaritons (SPPs) in dye doped polymeric microcylinder cavities deposited onto gold and silver wires. The stimulated emission spectra featured a characteristic series of laser modes, with modal spacing corresponding to SPPs propagating at the interface between the metal and dielectric. A plasmonic microlaser adds to the toolbox of plasmonic devices and plasmonic metamaterials and enables on-chip plasmonic generation and loss compensation. PMID- 21635089 TI - Low-power laser deformation of an air-liquid interface. AB - We report on the deformation of an air-water surface with a totally reflected low power laser beam, inducing a convex mirror effect on the beam propagation. This bending is stronger close to the critical angle and depends on the polarization of the laser light. A model, leading to a simple dependence between the Goos Hanchen shift and the radius of curvature of the interface, supports these observations. Bendings with radius of curvature as low as 0.10 m are demonstrated. PMID- 21635090 TI - General mechanism for the meandering instability of rivulets of Newtonian fluids. AB - A rivulet flowing down an inclined plane often does not follow a straight path, but starts to meander spontaneously. Here we show that this instability is the result of two key ingredients: fluid inertia and anisotropy of the friction between rivulet and substrate. Meandering only occurs if the motion normal to the instantaneous flow direction is more difficult than parallel to it. We give a quantitative criterion for the onset of meandering and confirm it by comparing to the flow of a rivulet between two glass plates which are wetted completely. Above the threshold, the rivulet follows an irregular pattern with a typical wavelength of a few cm. PMID- 21635091 TI - Energy dissipating structures produced by walls in two-dimensional flows at vanishing viscosity. AB - We perform numerical experiments of a dipole crashing into a wall, a generic event in two-dimensional incompressible flows with solid boundaries. The Reynolds number (Re) is varied from 985 to 7880, and no-slip boundary conditions are approximated by Navier boundary conditions with a slip length proportional to Re( 1). Energy dissipation is shown to first set up within a vorticity sheet of thickness proportional to Re(-1) in the neighborhood of the wall, and to continue as this sheet rolls up into a spiral and detaches from the wall. The energy dissipation rate integrated over these regions appears to converge towards Re independent values, indicating the existence of energy dissipating structures that persist in the vanishing viscosity limit. PMID- 21635092 TI - Vortex formation in a shock-accelerated gas induced by particle seeding. AB - An instability forms in gas of constant density (air) with an initial nonuniform seeding of small particles or droplets as a planar shock wave passes through the two-phase medium. The seeding nonuniformity is produced by vertical injection of a slow-moving jet of air premixed with glycol droplets or smoke particles into the test section of a shock tube, with the plane of the shock parallel to the axis of the jet. After the shock passage, two counterrotating vortices form in the plane normal to that axis. The physical mechanism of the instability we observe is peculiar to multiphase flow, where the shock acceleration causes the second (embedded) phase to move with respect to the embedding medium. With sufficient seeding concentration, this leads to entrainment of the embedding phase that acquires a relative velocity dependent on the initial seeding, resulting in vortex formation in the flow. PMID- 21635093 TI - Coherent oscillations of turbulent Rayleigh-Benard convection in a thin vertical disk. AB - A well-defined oscillation is observed in the power spectrum of several fluctuating signals in turbulent Rayleigh-Benard convection occurring in a thin vertical disk filled with water. The experiment reveals that the coherent oscillations are produced by periodic emission of thermal plumes, which gives rise to periodic pulses of forcing, resulting in a pulsed large-scale circulation in the thin cell. The experimental results agree well with the theoretical predictions made from two coupled nonlinear delayed equations. PMID- 21635094 TI - Nonlinear longitudinal space charge oscillations in relativistic electron beams. AB - In this Letter we study the evolution of an initial periodic modulation in the temporal profile of a relativistic electron beam under the effect of longitudinal space-charge forces. Linear theory predicts a periodic exchange of the modulation between the density and the energy profiles at the beam plasma frequency. For large enough initial modulations, wave breaking occurs after 1/2 period of plasma oscillation leading to the formation of short current spikes. We confirm this effect by direct measurements on a ps-modulated electron beam from an rf photoinjector. These results are useful for the generation of intense electron pulse trains for advanced accelerator applications. PMID- 21635084 TI - Measurements of direct CP violating asymmetries in charmless decays of strange bottom mesons and bottom baryons. AB - We report measurements of direct CP-violating asymmetries in charmless decays of neutral bottom hadrons to pairs of charged hadrons with the upgraded Collider Detector at the Fermilab Tevatron. Using a data sample corresponding to 1 fb(-1) of integrated luminosity, we obtain the first measurements of direct CP violation in bottom strange mesons, A(CP)(B(s)(0)->K(-)pi(+))=+0.39+/-0.15(stat)+/ 0.08(syst), and bottom baryons, A(CP)(Lambda(b)(0)->ppi(-))=+0.03+/-0.17(stat)+/ 0.05(syst) and A(CP)(Lambda(b)(0)->pK(-))=+0.37+/-0.17(stat)+/-0.03(syst). In addition, we measure CP violation in B(0)->K(+)pi(-) decays with 3.5sigma significance, A(CP)(B(0)->K(+)pi(-))=-0.086+/-0.023(stat)+/-0.009(syst), in agreement with the current world average. Measurements of branching fractions of B(s)(0)->K(+)K(-) and B(0)->pi(+)pi(-) decays are also updated. PMID- 21635095 TI - Spontaneous electromagnetic emission from a strongly localized plasma flow. AB - Laboratory observations of electromagnetic ion-cyclotron waves generated by a localized transverse dc electric field are reported. Experiments indicate that these waves result from a strong E*B flow inhomogeneity in a mildly collisional plasma with subcritical magnetic field-aligned current. The wave amplitude scales with the magnitude of the applied radial dc electric field. The electromagnetic signatures become stronger with increasing plasma beta, and the radial extent of the power is larger than that of the electrostatic counterpart. Near-Earth space weather implications of the results are discussed. PMID- 21635096 TI - Controlling the spacing of attosecond pulse trains from relativistic surface plasmas. AB - When a laser pulse hits a solid surface with relativistic intensities, XUV attosecond pulses are generated in the reflected light. We present an experimental and theoretical study of the temporal properties of attosecond pulse trains in this regime. The recorded harmonic spectra show distinct fine structures which can be explained by a varying temporal pulse spacing that can be controlled by the laser contrast. The pulse spacing is directly related to the cycle-averaged motion of the reflecting surface. Thus the harmonic spectrum contains information on the relativistic plasma dynamics. PMID- 21635097 TI - Measurements of electron transport in foils irradiated with a picosecond time scale laser pulse. AB - The heating of solid foils by a picosecond time scale laser pulse has been studied by using x-ray emission spectroscopy. The target material was plastic foil with a buried layer of a spectroscopic tracer material. The laser pulse length was either 0.5 or 2 ps, which resulted in a laser irradiance that varied over the range 10(16)-10(19) W/cm(2). Time-resolved measurements of the buried layer emission spectra using an ultrafast x-ray streak camera were used to infer the density and temperature conditions as a function of laser parameters and depth of the buried layer. Comparison of the data to different models of electron transport showed that they are consistent with a model of electron transport that predicts the bulk of the target heating is due to return currents. PMID- 21635098 TI - Effect of lattice structure on energetic electron transport in solids irradiated by ultraintense laser pulses. AB - The effect of lattice structure on the transport of energetic (MeV) electrons in solids irradiated by ultraintense laser pulses is investigated using various allotropes of carbon. We observe smooth electron transport in diamond, whereas beam filamentation is observed with less ordered forms of carbon. The highly ordered lattice structure of diamond is shown to result in a transient state of warm dense carbon with metalliclike conductivity, at temperatures of the order of 1-100 eV, leading to suppression of electron beam filamentation. PMID- 21635099 TI - Dynamics of dark solitons in a trapped superfluid fermi gas. AB - We study soliton oscillations in a trapped superfluid Fermi gas across the Bose Einstein condensate to Bardeen-Cooper-Schrieffer (BEC-BCS) crossover. We derive an exact equation for the oscillation period in terms of observable quantities, which we confirm by solving the time-dependent Bogoliubov-de Gennes equations. Hence we reveal the appearance and dynamics of solitons across the crossover, and show that the period dramatically increases as the soliton becomes shallower on the BCS side of the resonance. Finally, we propose an experimental protocol to test our predictions. PMID- 21635100 TI - Pair superfluidity of three-body constrained bosons in two dimensions. AB - We examine the equilibrium properties of lattice bosons with attractive on-site interactions in the presence of a three-body hard-core constraint that stabilizes the system against collapse and gives rise to a dimer superfluid phase. Employing quantum Monte Carlo simulations, the ground state phase diagram of this system on the square lattice is analyzed. In particular, we study the quantum phase transition between the atomic and dimer superfluid regime and analyze the nature of the superfluid-insulator transitions. Evidence is provided for the existence of a tricritical point along the saturation transition line, where the transition changes from being first order to a continuous transition of the dilute Bose gas of holes. The Berzinskii-Kosterlitz-Thouless transition from the dimer superfluid to the normal fluid is found to be consistent with an anomalous stiffness jump, as expected from the unbinding of half-vortices. PMID- 21635101 TI - NMR study of the dynamics of 3He impurities in the proposed supersolid state of solid 4He. AB - The dynamics of (3)He atoms in solid (4)He have been investigated by measuring the NMR relaxation times T(1) and T(2) in the region where a significant nonclassical rotational inertia fraction has been reported. For (3)He concentrations x(3)=16 and 24 ppm, changes are observed for both the spin-lattice relaxation time T(1) and the spin-spin relaxation time T(2) at the temperatures corresponding to the onset of the nonclassical rotational inertia fraction and, at lower temperatures, to the (3)He-(4)He phase separation. The magnitudes of T(1) and T(2) at temperatures above the phase separation agree roughly with existing theory based on the tunneling of (3)He impurities in the elastic strain field due to isotopic mismatch. However, a distinct peak in T(1) and a less well resolved feature in T(2) are observed near the reported nonclassical rotational inertia fraction onset temperature, in contrast to the temperature-independent relaxation times predicted by the tunneling theory. PMID- 21635102 TI - Periodic nanostructures: spatial dispersion mimics chirality. AB - Polarization rotation in isotropic materials is commonly associated with chirality, i.e., structures with a handedness which are not identical with their mirror image. We observe this effect in the visible and near-IR regions at oblique incidence in the optical response of a subwavelength square array of holes. Mapping the complete k space via Mueller-matrix spectroscopic ellipsometry, we find that in specific directions the rotary power is orders of magnitude larger than that observed for chiral molecules. Although experimentally indistinguishable, the physics behind the two phenomena is fundamentally different: While optical activity is a consequence of magnetic interactions, nanostructures on a square lattice rotate the polarization due to spatial dispersion. PMID- 21635103 TI - Weak intermolecular interactions in an ionically bound molecular adsorbate: cyclopentadienyl/Cu(111). AB - The dissociative adsorption of cyclopentadiene (C(5)H(6)) on Cu(111) yields a cyclopentadienyl (Cp) species with strongly anionic characteristics. The Cp potential energy surface and frictional coupling to the substrate are determined from measurements of dynamics of the molecule together with density functional calculations. The molecule is shown to occupy degenerate threefold adsorption sites and molecular motion is characterized by a low diffusional energy barrier of 40+/-3 meV with strong frictional dissipation. Repulsive dipole-dipole interactions are not detected despite charge transfer from substrate to adsorbate. PMID- 21635104 TI - Role of dispersion forces in the structure of graphene monolayers on Ru surfaces. AB - Elaborate density functional theory (DFT) calculations that include the effect of van der Waals (vdW) interactions have been carried out for graphene epitaxially grown on Ru(0001). The calculations predict a reduction of structural corrugation in the observed moire pattern of about 25% (~0.4 A) with respect to DFT calculations without vdW corrections. The simulated STM topographies are close to the experimental ones in a wide range of bias voltage around the Fermi level. PMID- 21635105 TI - Deformation of an elastic substrate by a three-phase contact line. AB - Young's classic analysis of the equilibrium of a three-phase contact line ignores the out-of-plane component of the liquid-vapor surface tension. While it is expected that this unresolved force is balanced by the elastic response of the solid, a definitive analysis has remained elusive because of an apparent divergence of stress at the contact line. While a number of theories have been presented to cut off the divergence, none of them have provided reasonable agreement with experimental data. We measure surface and bulk deformation of a thin elastic film near a three-phase contact line using fluorescence confocal microscopy. The out-of-plane deformation is well fit by a linear elastic theory incorporating an out-of-plane restoring force due to the surface tension of the solid substrate. This theory predicts that the deformation profile near the contact line is scale-free and independent of the substrate elastic modulus. PMID- 21635106 TI - Atomic-scale friction on stepped surfaces of ionic crystals. AB - We report on high-resolution friction force microscopy on a stepped NaCl(001) surface in ultrahigh vacuum. The measurements were performed on single cleavage step edges. When blunt tips are used, friction is found to increase while scanning both up and down a step edge. With atomically sharp tips, friction still increases upwards, but it decreases and even changes sign downwards. Our observations extend previous results obtained without resolving atomic features and are associated with the competition between the Schwobel barrier and the asymmetric potential well accompanying the step edges. PMID- 21635107 TI - Mott transition in modulated lattices and parent insulator of (K,Tl)(y)Fe(x)Se(2) superconductors. AB - The degree of electron correlations remains a central issue in the iron-based superconductors. The parent iron pnictides are antiferromagnetic, and their bad metal behavior has been interpreted in terms of proximity to a Mott transition. We study such a transition in multiorbital models on modulated lattices containing an ordered pattern of iron vacancies, using a slave-rotor method. We show that the ordered vacancies lead to a band narrowing, which pushes the system to the Mott insulator side. This effect is proposed to underlie the insulating behavior observed in the parent compounds of the newly discovered (K,Tl)(y)Fe(x)Se(2) superconductors. PMID- 21635108 TI - Fingerprints of intrinsic phase separation: magnetically doped two-dimensional electron gas. AB - In addition to Anderson and Mott localization, intrinsic phase separation has long been advocated as the third fundamental mechanism controlling the doping driven metal-insulator transitions. In electronic system, where charge neutrality precludes global phase separation, it may lead to various inhomogeneous states and dramatically affect transport. Here we theoretically predict the precise experimental signatures of such phase separation-driven metal-insulator transitions. We show that anomalous transport is expected in an intermediate regime around the transition, displaying very strong temperature and magnetic field dependence but very weak density dependence. Our predictions find striking agreement with recent experiments on Mn-doped CdTe quantum wells, a system where we identify the microscopic origin for intrinsic phase separation. PMID- 21635109 TI - Uncovering the complex behavior of hydrogen in Cu2O. AB - The behavior of hydrogen in p-type Cu(2)O has been reported to be quite unusual. Muon experiments have been unable to ascertain the preferential hydrogen site within the Cu(2)O lattice, and indicate that hydrogen causes an electrically active level near the middle of the band gap, whose nature, whether accepting or donating, is not known. In this Letter, we use screened hybrid-density-functional theory to study the nature of hydrogen in Cu(2)O, and identify for the first time the "quasiatomic" site adopted by hydrogen in Cu(2)O. We show that hydrogen will always act as a hole killer in p-type Cu(2)O, and is one likely cause of the low performance of Cu(2)O solar cell devices. PMID- 21635110 TI - Ferromagnetic enhancement of CE-type spin ordering in (Pr,Ca)MnO3. AB - We present resonant soft x-ray scattering results from small bandwidth manganites (Pr,Ca)MnO(3), which show that the CE-type spin ordering (SO) at the phase boundary is stabilized only below the canted antiferromagnetic transition temperature and enhanced by ferromagnetism in the macroscopically insulating state (FM-I). Our results reveal the fragility of the CE-type ordering that underpins the colossal magnetoresistance effect in this system, as well as an unexpected cooperative interplay between FM-I and CE-type SO which is in contrast to the competitive interplay between the ferromagnetic metallic state and CE-type ordering. PMID- 21635111 TI - Pressure-induced valence crossover in superconducting CeCu2Si2. AB - Measurement of the Ce valence in the heavy fermion CeCu(2)Si(2) is reported for the first time under pressure and at low temperature (T=14 K) in proximity of the superconducting region. CeCu(2)Si(2) is considered as a strong candidate for a new type of pairing mechanism related to critical valence fluctuations which could set in at high pressure in the vicinity of the second superconducting dome. A quantitative estimate of the valence in this pressure region was achieved from the measurements of the Ce L(3) edge in the high-resolution partial-fluorescence yield mode and subsequent analysis of the spectra within the Anderson impurity model. While a clear increase of the Ce valence is found, the weak electron transfer and the continuous valence change under pressure suggests a crossover regime with the hypothetical valence line terminating at a critical end point T(cr) close to zero. PMID- 21635112 TI - Semiclassical neutral atom as a reference system in density functional theory. AB - We use the asymptotic expansions of the semiclassical neutral atom as a reference system in density functional theory to construct accurate generalized gradient approximations (GGAs) for the exchange-correlation and kinetic energies without any empiricism. These asymptotic functionals are among the most accurate GGAs for molecular systems, perform well for solid state, and overcome current GGA state of the art in frozen density embedding calculations. Our results also provide evidence for the conjointness conjecture between exchange and kinetic energies of atomic systems. PMID- 21635113 TI - Coherent heavy quasiparticles in a CePt5 surface alloy. AB - We report on the results of a high-resolution angle-resolved photoemission study on the ordered surface alloy CePt(5). The temperature dependence of the spectra show the formation of the coherent low-energy heavy-fermion band near the Fermi level. These experimental data are supported by a multiband model calculation in the framework of the dynamical mean-field theory. PMID- 21635114 TI - Spin-seebeck effect: a phonon driven spin distribution. AB - Here we report on measurements of the spin-Seebeck effect in GaMnAs over an extended temperature range alongside the thermal conductivity, specific heat, magnetization, and thermoelectric power. The amplitude of the spin-Seebeck effect in GaMnAs scales with the thermal conductivity of the GaAs substrate and the phonon-drag contribution to the thermoelectric power of the GaMnAs, demonstrating that phonons drive the spin redistribution. A phenomenological model involving phonon-magnon drag explains the spatial and temperature dependence of the measured spin distribution. PMID- 21635115 TI - Slow conductance relaxation in insulating granular Al: evidence for screening effects. AB - It is shown that the conductance relaxations observed in electrical field effect measurements on granular Al films are the sum of two contributions. One is sensitive to gate voltage changes and gives the already reported anomalous electrical field effect. The other one is independent of the gate voltage history and starts when the films are cooled down to low temperature. Their relative amplitude is strongly thickness-dependent which demonstrates the existence of a finite screening length in our insulating films and allows its quantitative estimate (about 10 nm at 4 K). This metalliclike screening should be taken into account in the electron glass models of disordered insulators. PMID- 21635116 TI - Unconventional transport in the "hole" regime of a si double quantum dot. AB - Studies of electronic charge transport through semiconductor double quantum dots rely on a conventional "hole" model of transport in the three-electron regime. We show that experimental measurements of charge transport through a Si double quantum dot in this regime cannot be fully explained using the conventional picture. Using a Hartree-Fock (HF) formalism and relevant HF energy parameters extracted from transport data in the multiple-electron regime, we identify a novel spin-flip cotunneling process that lifts a singlet blockade. PMID- 21635117 TI - Numerical calculation of the neutral fermion gap at the nu=5/2 fractional quantum Hall state. AB - We present the first numerical computation of the neutral fermion gap, Delta(F), in the nu=5/2 quantum Hall state, which is analogous to the energy gap for a Bogoliubov-de Gennes quasiparticle in a superconductor. We find Delta(F)~0.027e(2)/epsilonl(0), comparable to the charge gap. We also deduce an effective Fermi velocity v(F) for neutral fermions from the low-energy spectra for odd numbers of electrons, and thereby obtain a correlation length xi(F)=v(F)/Delta(F)~1.3l(0). We comment on implications for experiments, topological quantum information processing, and electronic mechanisms of superconductivity. PMID- 21635118 TI - Cooling of nanomechanical resonators by thermally activated single-electron transport. AB - We show that the vibrations of a nanomechanical resonator can be cooled to near its quantum ground state by tunneling injection of electrons from a scanning tunneling microscope tip. The interplay between two mechanisms for coupling the electronic and mechanical degrees of freedom results in a bias-voltage-dependent difference between the probability amplitudes for vibron emission and absorption during tunneling. For a bias voltage just below the Coulomb blockade threshold, we find that absorption dominates, which leads to cooling corresponding to an average vibron population of the fundamental bending mode of 0.2. PMID- 21635119 TI - Absence of a holelike fermi surface for the iron-based K0.8F1.7Se2 superconductor revealed by angle-resolved photoemission spectroscopy. AB - We have performed an angle-resolved photoemission spectroscopy study of the new iron-based superconductor K(0.8)Fe(1.7)Se(2) (T(c)~30 K). Clear band dispersion is observed with the overall bandwidth renormalized by a factor of 2.5 compared to our local density approximation calculations, indicating relatively strong correlation effects. Only an electronlike band crosses the Fermi energy, forming a nearly circular Fermi surface (FS) at M (pi, 0). The holelike band at Gamma sinks ~90 meV below the Fermi energy, with an indirect band gap of 30 meV, to the bottom of the electronlike band. The observed FS topology in this superconductor favors (pi, pi) inter-FS scattering between the electronlike FSs at the M points, in sharp contrast to other iron-based superconductors which favor (pi, 0) inter FS scattering between holelike and electronlike FSs. PMID- 21635120 TI - Electronic structure of superconducting KC8 and nonsuperconducting LiC6 graphite intercalation compounds: evidence for a graphene-sheet-driven superconducting state. AB - We have performed photoemission studies of the electronic structure in LiC(6) and KC(8), a nonsuperconducting and a superconducting graphite intercalation compound, respectively. We have found that the charge transfer from the intercalant layers to graphene layers is larger in KC(8) than in LiC(6), opposite of what might be expected from their chemical composition. We have also measured the strength of the electron-phonon interaction on the graphene-derived Fermi surface to carbon derived phonons in both materials and found that it follows a universal trend where the coupling strength and superconductivity monotonically increase with the filling of graphene pi(*) states. This correlation suggests that both graphene-derived electrons and graphene-derived phonons are crucial for superconductivity in graphite intercalation compounds. PMID- 21635121 TI - Mechanism for explaining differences in the order parameters of FeAs-based and FeP-based pnictide superconductors. AB - We put forward a scenario that explains the difference between the order parameter character in arsenide (As) and phosphorous (P) iron-based superconductors. Using functional renormalization group to analyze it in detail, we find that nodal superconductivity on the electron pockets (hole pocket gaps are always nodeless) can naturally appear when the hole pocket at (pi,pi) in the unfolded Brillouin zone is absent, as is the case in LaOFeP. There, electron electron interactions render the gap on the electron pockets softly nodal (of s(+/-) form). When the pocket of d(xy) orbital character is present, intraorbital interactions with the d(xy) part of the electron Fermi surface drives the superconductivity nodeless. PMID- 21635122 TI - Evolution of Kondo resonance from a single impurity molecule to the two dimensional lattice. AB - The successive spectral evolution of the Kondo resonance state was investigated from a single iron(II) phthalocyanine molecule to the two-dimensional lattice on Au(111) by interrogating the individual molecules with a scanning tunneling microscope. A sharp Kondo peak appears in the single-impurity regime, which broadens and splits as the lattice builds up. The origin of spectral evolution together with the electronic ground state of the lattice are discussed based on the competition of the Kondo effect and Rudermann-Kittel-Kasuya-Yosida coupling between the molecular spins. PMID- 21635030 TI - Measurement of dijet azimuthal decorrelations in pp collisions at sqrt(s)=7 TeV. AB - Azimuthal decorrelations between the two central jets with the largest transverse momenta are sensitive to the dynamics of events with multiple jets. We present a measurement of the normalized differential cross section based on the full data set (?Ldt=36 pb(-1)) acquired by the ATLAS detector during the 2010 sqrt(s)=7 TeV proton-proton run of the LHC. The measured distributions include jets with transverse momenta up to 1.3 TeV, probing perturbative QCD in a high-energy regime. PMID- 21635123 TI - Rods of neutron scattering intensity in Yb2Ti2O7: compelling evidence for significant anisotropic exchange in a magnetic pyrochlore oxide. AB - Paramagnetic correlations in the magnetic material Yb(2)Ti(2)O(7) have been investigated via neutron scattering, revealing a [111] rod of scattering intensity. Assuming interactions between the Yb(3+) ions composed of all symmetry allowed nearest neighbor exchange interactions and long-range dipolar interactions, we construct a model Hamiltonian that allows for an excellent description of the neutron scattering data. Our results provide compelling evidence for significant anisotropic exchange interactions in an insulating magnetic pyrochlore oxide. We also compute the real space correlations leading to the [111] rod of scattering. PMID- 21635126 TI - Blueshifting the onset of optical UV absorption for water under pressure. AB - First-principles calculations show that the optical UV absorption onset of solid water is blueshifted with increasing pressure. Across several crystal structures and a wide pressure range, the optical gap increases almost linearly with external pressure, making solid water more transparent. The origin of this unusual effect can be traced back to an increased Stark shift caused by water's electrostatic environment at smaller volumes. PMID- 21635124 TI - Near-unity quantum yields of biexciton emission from CdSe/CdS nanocrystals measured using single-particle spectroscopy. AB - Biexciton photoluminescence (PL) quantum yields (Q(2X)) of individual CdSe/CdS core-shell nanocrystal quantum dots with various shell thicknesses are derived from independent PL saturation and two-photon correlation measurements. We observe a near-unity Q(2X) for some nanocrystals with an ultrathick 19-monolayer shell. High Q(2X)'s are, however, not universal and vary widely among nominally identical nanocrystals indicating a significant dependence of Q(2X) upon subtle structural differences. Interestingly, our measurements indicate that high Q(2X)'s are not required to achieve complete suppression of PL intensity fluctuations in individual nanocrystals. PMID- 21635125 TI - Renormalization of optical excitations in molecules near a metal surface. AB - The lowest electronic excitations of benzene and a set of donor-acceptor molecular complexes are calculated for the gas phase and on the Al(111) surface using the many-body Bethe-Salpeter equation. The energy of the charge-transfer excitations obtained for the gas phase complexes are found to be around 10% lower than the experimental values. When the molecules are placed outside the surface, the enhanced screening from the metal reduces the exciton binding energies by several eVs and the transition energies by up to 1 eV depending on the size of the transition-generated dipole. As a striking consequence we find that close to the metal surface the optical gap of benzene can exceed its quasiparticle gap. A classical image charge model for the screened Coulomb interaction can account for all these effects which, on the other hand, are completely missed by standard time-dependent density functional theory. PMID- 21635127 TI - Electrical detection of coherent nuclear spin oscillations in phosphorus-doped silicon using pulsed ENDOR. AB - We demonstrate the electrical detection of pulsed X-band electron nuclear double resonance (ENDOR) in phosphorus-doped silicon at 5 K. A pulse sequence analogous to Davies ENDOR in conventional electron spin resonance is used to measure the nuclear spin transition frequencies of the (31)P nuclear spins, where the (31)P electron spins are detected electrically via spin-dependent transitions through Si/SiO(2) interface states, thus not relying on a polarization of the electron spin system. In addition, the electrical detection of coherent nuclear spin oscillations is shown, demonstrating the feasibility to electrically read out the spin states of possible nuclear spin qubits. PMID- 21635128 TI - Determination of viral capsid elastic properties from equilibrium thermal fluctuations. AB - We apply two-dimensional elasticity theory to viral capsids to develop a framework for calculating elastic properties of viruses from equilibrium thermal fluctuations of the capsid surface in molecular dynamics and elastic network model trajectories. We show that the magnitudes of the long wavelength modes of motion available in a simulation with all atomic degrees of freedom are recapitulated by an elastic network model. For the mode spectra to match, the elastic network model must be scaled appropriately by a factor which can be determined from an icosahedrally constrained all-atom simulation. With this method we calculate the two-dimensional Young's modulus Y, bending modulus kappa, and Foppl-von Karman number gamma, for the T=1 mutant of the Sesbania mosaic virus. The values determined are in the range of previous theoretical estimates. PMID- 21635129 TI - Depletion-induced structure and dynamics in bimodal colloidal suspensions. AB - Combined small angle x-ray scattering and x-ray photon correlation spectroscopy studies of moderately concentrated bimodal hard-sphere colloidal suspensions in the fluid phase show that depletion-induced demixing introduces spatially heterogeneous dynamics with two distinct time scales. The adhesive nature, as well as the mobility, of the large particles is determined by the level of interaction within the monomodal domains. This interaction is driven by osmotic forces, which are governed by the relative concentration of the constituents. PMID- 21635130 TI - Axial thermal rotation of slender rods. AB - Axial rotational diffusion of rodlike polymers is important in processes such as microtubule filament sliding and flagella beating. By imaging the motion of small kinks along the backbone of chains of DNA-linked colloids, we produce a direct and systematic measurement of axial rotational diffusivity of rods both in bulk solution and near a wall. The measured diffusivities decrease linearly with the chain length, irrespective of the distance from a wall, in agreement with slender body hydrodynamics theory. Moreover, the presence of small kinks does not affect the chain's axial diffusivity. Our system and measurements provide insights into fundamental axial diffusion processes of slender objects, which encompass a wide range of entities including biological filaments and linear polymer chains. PMID- 21635131 TI - Evidence of negative-index refraction in nonlinear chemical waves. AB - The negative index of refraction of nonlinear chemical waves has become a recent focus in nonlinear dynamics researches. Theoretical analysis and computer simulations have predicted that the negative index of refraction can occur on the interface between antiwaves and normal waves in a reaction-diffusion (RD) system. However, no experimental evidence has been found so far. In this Letter, we report our experimental design in searching for such a phenomenon in a chlorite iodide-malonic acid (CIMA) reaction. Our experimental results demonstrate that competition between waves and antiwaves at their interface determines the fate of the wave interaction. The negative index of refraction was only observed when the oscillation frequency of a normal wave is significantly smaller than that of the antiwave. All experimental results were supported by simulations using the Lengyel-Epstein RD model which describes the CIMA reaction-diffusion system. PMID- 21635132 TI - Extreme events on complex networks. AB - A wide spectrum of extreme events ranging from traffic jams to floods take place on networks. Motivated by these, we employ a random walk model for transport and obtain analytical and numerical results for the extreme events on networks. They reveal an unforeseen, and yet a robust, feature: small degree nodes of a network are more likely to encounter extreme events than the hubs. Further, we also study the recurrence time distribution and scaling of the probabilities for extreme events. These results suggest a revision of design principles and can be used as an input for designing the nodes of a network so as to smoothly handle extreme events. PMID- 21635133 TI - DNA-based computing of strategic assignment problems. AB - DNA-based computing is a novel technique to tackle computationally difficult problems, in which computing time grows exponentially corresponding to problematic size. A strategic assignment problem is a typical nondeterministic polynomial problem, which is often associated with strategy applications. In this Letter, a new approach dealing with strategic assignment problems is proposed based on manipulating DNA strands, which is believed to be better than the conventional silicon-based computing in solving the same problem. PMID- 21635134 TI - Comment on "optical orbital angular momentum from the curl of polarization". PMID- 21635136 TI - Assessment of dyspnea in asthma: validation of The Dyspnea-12. AB - BACKGROUND: Dyspnea is a prominent symptom in asthma. The Dyspnea-12 (D-12), an instrument that quantifies breathlessness using 12 descriptors that tap the physical and affective aspects, has shown promise for the measurement of dyspnea in cardiorespiratory disease. OBJECTIVE: We report the results of a study designed to test the validity and reliability of the D-12 in a population of patients with asthma. METHODS: This cross-sectional study included 102 patients with asthma. Subjects completed the D-12, Hospital Anxiety and Depression scale, St. George's Respiratory Questionnaire (SGRQ), and Medical Research Council scale. Confirmatory factor analysis confirmed the two-component structure of the D-12 (i.e., seven items that tap the physical aspects of breathlessness and five items that tap the affective aspects). RESULTS: The D-12 subscales had excellent internal reliability (Cronbach's alpha for the "physical" score was 0.94 and the affective score was 0.95). The D-12 physical component was more strongly correlated with SGRQ Symptoms (r = 0.648), SGRQ Activities (r = 0.635) and Medical Research Council grade (r = 0.636), while the affective component was more strongly correlated with SGRQ Impacts (r = 0.765) and Hospital Anxiety and Depression scale scores (anxiety r = 0.641 and depression r = 0.602). CONCLUSION: This study supports validity of the D-12 for use in the assessment of dyspnea of patients with asthma. It assesses one of the most pertinent symptoms of asthma from two viewpoints-physical and affective. PMID- 21635137 TI - Preparation and characterization of lung-targeting ceftiofur-loaded gelatin microspheres. AB - BACKGROUND: Ceftiofur is an effective antibiotic against respiratory infections in livestock. However, ceftiofur concentration that is found in lungs after intravenous injection is not effective. Fortunately, ceftiofur-loaded gelatin microsphere (Cef-MS) enjoys advantages of lung-targeting and can achieve an effective concentration. However, no study has been reported on this modality of drug delivery. OBJECTIVE: We investigated the properties of this delivery modality--lung targeting ceftiofur-Cef-MSs. METHODS: We prepared Cef-MS and investigated drug loading, stability and release characteristics in vitro and studied tissue distribution patterns and potential lung injury in mice. RESULTS: Results showed that the average size and span value are 21.26 MUm and 1.07, respectively. Drug loading and loading efficiency were 15.31 and 76.55%, respectively. Cef-MSs were stable in light, heat and humidity, except that agglutinative phenomenon was observed in 90% humidity after 10 days. Cef-MS presented a slower in vitro release pattern compared to ceftiofur. Cef-MS mainly concentrates in lungs after intravenous administration. Furthermore, histopathological studies showed that Cef-MS only induces mild and reversible lung injury and is biologically safe. CONCLUSION: Cef-MS is a promising alternative form with high lung-targeting properties for the treatment of respiratory infections. PMID- 21635138 TI - Safety assessment of thiolated polymers: effect on ciliary beat frequency in human nasal epithelial cells. AB - OBJECTIVE: The aim of this study was to investigate the nasal safety of gel formulations of thiolated polymers (thiomers) by assessing their effect on ciliary beat frequency (CBF) in human nasal epithelial cells. METHODS: Poly(acrylic acid) 450 kDa-cysteine (PAA-cys) and alginate-cysteine (alg-cys) were synthesized by covalent attachment of L-cysteine to the polymeric backbone. The cationic polymer chitosan-thiobutylamidine (chito-TBA) was synthesized by attaching iminothiolane to chitosan. CBF using was measured by a photometric system. CBF was measured before incubating the cells with test gels, during incubation and after washing out the polymeric test gels to evaluate reversibility of cilio-inhibition. The influence of viscosity on CBF was determined by using hydroxyethylcellulose (HEC)-gels of various concentrations. RESULTS: Ciliary beating was observed to be affected by viscosity, but cilia were still beating in the presence of a HEC-gel displaying an apparent viscosity of 25 Pa.s. In case of thiolated polymers and their unmodified control, a concentration dependent decrease in CBF could be observed. PAA-cys, alg-cys, chito-TBA and their corresponding unmodified controls exhibited a moderate cilio-inhibitory effect, followed by a partial recovery of CBF when used at a concentration of 1%. Alg-cys 2% and chito-TBA 2% (m/v) gels exhibited severe cilio-inhibition, which was partially reversible. L-cysteine and reduced glutathione led to mild cilio inhibition at concentrations of 3% (m/v). CONCLUSIONS: Taking into account that dilution after application and cilio-modifying effects is usually more pronounced under in vitro conditions, thiomers can be considered as suitable excipients for nasal drug delivery systems. PMID- 21635139 TI - The Botrytis cinerea Reg1 protein, a putative transcriptional regulator, is required for pathogenicity, conidiogenesis, and the production of secondary metabolites. AB - Botrytis cinerea, which causes gray-mold rot, attacks a wide range of plant species. To understand the infection process, the role of a putative transcriptional regulator, BcReg1 (regulator 1), in pathogenicity was studied. This transcriptional regulator shows similarity to the morphological switch regulators Candida albicans Wor1 and Histoplasma capsulatum Ryp1. Gene knock-out and complementation studies revealed that bcreg1 is required for pathogenicity. The bcreg1 mutant is able to penetrate plant tissue but is not able to cause necrotic lesions. In addition, the mutant is blocked in conidia formation and does not produce detectable levels of the sesquiterpene botrydial and the polyketide botcinic acid. Based on transcript expression levels, it can be concluded that bcreg1 is a downstream target of two mitogen-activated protein kinases, BcSak1 and Bmp3. PMID- 21635140 TI - Role of hydroxycinnamic acids in the infection of maize silks by Fusarium graminearum Schwabe. AB - In the current study, the hydroxycinnamic acids in silks of diverse maize inbred lines differing in Fusarium resistance were determined at several times after inoculation with Fusarium graminearum or sterile water as control. The main objective was to determine the possible relationship between the hydroxycinnamic acid changes in silks and ear rot resistance. Several changes in the cell-wall bound hydroxycinnamic acid concentrations were observed after inoculation with F. graminearum, although these changes were not directly correlated with genotypic resistance to this fungus. Ester-bound ferulic acid decreased, probably due to degradation of hemicellulose by hydrolytic enzymes produced by Fusarium spp., while p-coumaric acid and diferulates showed slight increases that, in conjunction, did not result in delayed F. graminearum progression through the silks. It is important to note that the decrease of ferulic acid in the F. graminearum treatment was faster in susceptible than in resistant genotypes, suggesting a differential hemicellulose degradation in silk tissues. Therefore, the ability of the maize genotypes to slow down that process through hemicellulose structural features or xylanase inhibitors needs to be addressed in future studies. PMID- 21635141 TI - The Fusarium virguliforme toxin FvTox1 causes foliar sudden death syndrome-like symptoms in soybean. AB - Fusarium virguliforme causes sudden death syndrome (SDS) in soybean. The pathogen has never been isolated from diseased foliar tissues; therefore, one or more toxins have been considered to cause foliar SDS development. Cell-free F. virguliforme culture filtrates containing a toxin causes foliar SDS in soybean. A low-molecular-weight protein of approximately 13.5 kDa (FvTox1), purified from F. virguliforme culture filtrates, produces foliar SDS-like symptoms in cut soybean seedlings. Anti-FvTox1 monoclonal antibodies raised against the purified FvTox1 were used in isolating the FvTox1 gene. In the presence of light, recombinant FvTox1 protein expressed in an insect cell line resulted in chlorosis and necrosis in soybean leaf disks that are typical foliar SDS symptoms. SDS susceptible but not the SDS-resistant soybean lines were sensitive to the baculovirus-expressed toxin. The requirement of light for foliar SDS-like symptom development indicates that FvTox1 induces foliar SDS in soybean, most likely through production of free radicals by interrupting photosynthesis. PMID- 21635142 TI - Pathogenicity, symptom development, and mycotoxin formation in wheat by Fusarium species frequently isolated from sugar beet. AB - Crop rotations with putative non-host crops such as sugar beet are often recommended to reduce Fusarium head blight (FHB) in cereals. However, recent observations have shown pathogenic, endophytic, and saprotrophic colonization of sugar beet with various Fusarium spp. Therefore, strains of seven species frequently isolated from sugar beet were tested for pathogenicity on wheat. Species-specific symptoms on heads and kernels were evaluated and the grains were analyzed for 20 mycotoxins with liquid chromatography-tandem mass spectrometry. Fusarium graminearum, F. culmorum, and F. cerealis from sugar beet caused typical FHB symptoms and mycotoxin contamination with deoxynivalenol and nivalenol, while a high incidence of black point was observed in heads inoculated with F. tricinctum or F. equiseti. Black point kernels revealed 3.4 to 14.5 times higher mycotoxin concentrations than symptomless grains, containing enniatin B1 at 38,000 MUg/kg, moniliformin at 4,900 MUg/kg, and 2-amino-14,16-dimethyloctadecan 3-ol at 5,500 MUg/kg, as well as monoacetoxyscirpenol at 2,600 MUg/kg and nivalenol at 3,800 MUg/kg. Monitoring of these latter two species in the field is hampered by the lack of typical head symptoms after infection. In further experiments, the impact of sugar beet residues on FHB severity and the correlation between mycotoxin contamination of cereal lots and the amount of black point have to be evaluated. PMID- 21635143 TI - Quantitative trait loci for adult-plant resistance to Mycosphaerella graminicola in two winter wheat populations. AB - Septoria tritici blotch (STB) is one of the most important leaf spot diseases in wheat worldwide. The goal of this study was to detect chromosomal regions for adult-plant resistance in large winter wheat populations to STB. Inoculation by two isolates with virulence to Stb6 and Stb15, both present in the parents, was performed and STB severity was visually scored plotwise as percent coverage of flag leaves with pycnidia-bearing lesions. 'Florett'/'Biscay' and 'Tuareg'/'Biscay', each comprising a cross of a resistant and a susceptible cultivar, with population sizes of 316 and 269 F(7:8) recombinant inbred lines, respectively, were phenotyped across four and five environments and mapped with amplified fragment length polymorphism, diversity array technology, and simple sequence repeat markers covering polymorphic regions of ~1,340 centimorgans. Phenotypic data revealed significant (P < 0.01) genotypic differentiation for STB, heading date, and plant height. Entry-mean heritabilities (h(2)) for STB were 0.73 for 'Florett'/'Biscay' and 0.38 for 'Tuareg'/'Biscay'. All correlations between STB and heading date as well as between STB and plant height were low (r = -0.13 to -0.20). In quantitative trait loci (QTL) analysis, nine and six QTL were found for STB ratings explaining, together, 55 and 51% of phenotypic variation in 'Florett'/'Biscay' and 'Tuareg'/'Biscay', respectively. Genotype environment and QTL-environment interactions had a large impact. Two major QTL were detected consistently across environments on chromosomes 3B and 6D from 'Florett' and chromosomes 4B and 6B from 'Tuareg', each explaining 12 to 17% of normalized adjusted phenotypic variance. These results indicate that adult-plant resistance to STB in both mapping populations was of a quantitative nature. PMID- 21635144 TI - Impact of the CYP3A4*1G polymorphism and its combination with CYP3A5 genotypes on tacrolimus pharmacokinetics in renal transplant patients. AB - AIM: Tacrolimus is a substrate of CYP3A4 and CYP3A5. The present study investigated the impact of the CYP3A4*1/*1G polymorphism compared with CYP3A5 genotypes on the dose-adjusted pharmacokinetics of tacrolimus. The effects of the polymorphism on the variability in tacrolimus pharmacokinetics among patients with the CYP3A5*1 allele (CYP3A5 expresser) and among those with CYP3A5*3/*3 genotype (nonexpresser) were also studied. MATERIALS & METHODS: A total of 136 renal allograft recipients were given repeated doses of tacrolimus every 12 h. On day 28 after the renal transplantation, blood tacrolimus concentrations were measured, and dose-adjusted pharmacokinetics were determined and compared with the corresponding genotype. RESULTS: The dose-adjusted AUC0-12 and C0 of tacrolimus were significantly lower in patients with the CYP3A4*1G allele and CYP3A5 expressers than those with the CYP3A4*1/*1 genotype and nonexpressers, respectively. In a multiple regression analysis, the dose-adjusted AUC0-12 and C0 values were associated with CYP3A4*1/*1 (p = 0.018 and 0.040, respectively) and CYP3A5*3/*3 (p < 0.001 each). The standardized regression coefficient for the AUC0-12 of tacrolimus was approximately twofold less for CYP3A4*1/*1 than CYP3A5*3/*3. The lowest dose-adjusted AUC0-12 was found in CYP3A5 expressers with the CYP3A4*1G allele. CONCLUSION: The CYP3A4*1/*1G polymorphism was associated with the pharmacokinetics of tacrolimus, however, its contribution to dose adjusted pharmacokinetics was approximately twofold less than that of the CYP3A5*1/*3 polymorphism. Although its effect on CYP3A4 activity is not clear, CYP3A4*1/*1G may be a candidate for a polymorphism affecting the interindividual variability in tacrolimus pharmacokinetics among CYP3A5 expressers. PMID- 21635145 TI - Prednisolone-induced changes in gene-expression profiles in healthy volunteers. AB - BACKGROUND: Prednisolone and other glucocorticoids (GCs) are potent anti inflammatory and immunosuppressive drugs. However, prolonged use at a medium or high dose is hampered by side effects of which the metabolic side effects are most evident. Relatively little is known about their effect on gene-expression in vivo, the effect on cell subpopulations and the relation to the efficacy and side effects of GCs. AIM: To identify and compare prednisolone-induced gene signatures in CD4+ T lymphocytes and CD14+ monocytes derived from healthy volunteers and to link these signatures to underlying biological pathways involved in metabolic adverse effects. MATERIALS & METHODS: Whole-genome expression profiling was performed on CD4+ T lymphocytes and CD14+ monocytes derived from healthy volunteers treated with prednisolone. Text-mining analyses was used to link genes to pathways involved in metabolic adverse events. RESULTS: Induction of gene expression was much stronger in CD4+ T lymphocytes than in CD14+ monocytes with respect to fold changes, but the number of truly cell-specific genes where a strong prednisolone effect in one cell type was accompanied by a total lack of prednisolone effect in the other cell type, was relatively low. Subsequently, a large set of genes was identified with a strong link to metabolic processes, for some of which the association with GCs is novel. CONCLUSION: The identified gene signatures provide new starting points for further study into GC-induced transcriptional regulation in vivo and the mechanisms underlying GC-mediated metabolic side effects. PMID- 21635146 TI - GALNT14 SNP as a potential predictor of response to combination chemotherapy using 5-FU, mitoxantrone and cisplatin in advanced HCC. AB - AIM: Despite the availability of targeted anticancer agents, combination chemotherapy remains an option for patients with far advanced hepatocellular carcinoma. We aimed to identify germline SNP markers in order to predict the objective response of combination chemotherapy using 5-fluorouracil, mitoxantrone and cisplatin (FMP). MATERIALS & METHODS: This study was conducted in two steps. First, a genome-wide retrospective screen was performed in a cohort of 16 patients. A candidate SNP marker was identified as being associated with the objective responses to the first course of FMP. Second, a validation of this candidate SNP was performed prospectively in an independent cohort of 41 patients. The survival curves were also compared in patient subgroups stratified by the SNP marker. RESULTS: The genome-wide screening revealed several candidate markers, including seven adjacent SNPs residing in a 5-kb linkage disequilibrium block and in an intronic region of GALNT14 on chromosome 2. Among them, the SNP rs9679162 manifested the strongest association when genotypic tests and kernel based association tests were performed. Significant association was found again between rs9679162 and the therapeutic responses in the validation cohort (p = 0.006326). A follow-up survival analysis demonstrated that patients with a homozygous rs9679162 genotype had better progression-free survival in both the retrospective and prospective cohorts (p = 0.00041 and 0.01485, respectively) than the other genotypes did. However, the overall survival curve is only different in the retrospective cohort (p = 0.00622) and not in the prospective cohort. CONCLUSION: The rs9679162 GALNT14 genotype is potentially associated with the objective response of the first course of FMP chemotherapy in patients with far advanced hepatocellular carcinoma. PMID- 21635147 TI - Novel CYP2C9 and VKORC1 gene variants associated with warfarin dosage variability in the South African black population. AB - AIM: Warfarin is a widely used therapeutic agent for long-term oral anticoagulation worldwide. Its administration is challenging owing to its narrow therapeutic range and serious adverse effects. Several environmental factors and numerous genes, of which CYP2C9 and VKORC1 are the most important, have been associated with interindividual dosage variability. Many studies have been conducted to understand warfarin dosage variability better, the majority of which have been focused on the Caucasian and African-American populations. Very little information is available regarding genetic influences of warfarin dosage variability in the South African black population. MATERIALS & METHODS: In this study, we genotyped 213 South African black individuals for CYP2C9 and VKORC1 variants and a small subset of environmental factors that may be responsible for warfarin dosage variability. RESULTS: We observed 26 novel SNPs and seven previously described CYP2C9 variants and three previously described but no novel VKORC1 SNPs. Only 11 of the CYP2C9 variants and two of the VKORC1 variants were observed at high enough allele frequencies to assess their impact on warfarin dosage. CONCLUSION: We demonstrate that CYP2C9*8 and two novel CYP2C9 SNPs (g.16179 and g.46028) are associated with a decrease in warfarin dosage, beta blockers are independently associated with a decrease in warfarin dosage and two known VKORC1 variants (rs7200749 and rs7294) are associated with an increase in warfarin dosage. The CYP2C9 and VKORC1 variants and a small subset of environmental factors used in the study explain approximately 45% of warfarin dosage variability in the South African black population. PMID- 21635148 TI - The use of molecular imaging of gene expression by radiotracers in gene therapy. AB - INTRODUCTION: Progress with gene-based therapies has been hampered by difficulties in monitoring the biodistribution and kinetics of vector-mediated gene expression. Recent developments in non-invasive imaging have allowed researchers and clinicians to assess the location, magnitude and persistence of gene expression in animals and humans. Such advances should eventually lead to improvement in the efficacy and safety of current clinical protocols for future treatments. AREAS COVERED: The molecular imaging techniques for monitoring gene therapy in the living subject, with a specific highlight on the key reporter gene approaches that have been developed and validated in preclinical models using the latest imaging modalities. The applications of molecular imaging to biotherapy, with a particular emphasis on monitoring of gene and vector biodistribution and on image-guided radiotherapy. EXPERT OPINION: Among the reporter gene/probe combinations that have been described so far, one stands out, in our view, as the most versatile and easy to implement: the Na/I symporter. This strategy, exploiting more than 50 years of experience in the treatment of differentiated thyroid carcinomas, has been validated in different types of experimental cancers and with different types of oncolytic viruses and is likely to become a key tool in the implementation of human gene therapy. PMID- 21635149 TI - Anthracycline cardiotoxicity. AB - INTRODUCTION: Anthracyclines are widely prescribed anticancer agents that cause a dose-related cardiotoxicity, often aggravated by nonanthracycline chemotherapeutics or new generation targeted drugs. Anthracycline cardiotoxicity may occur anytime in the life of cancer survivors. Understanding the molecular mechanisms and clinical correlates of cardiotoxicity is necessary to improve the therapeutic index of anthracyclines or to identify active, but less cardiotoxic analogs. AREAS COVERED: The authors review the pharmacokinetic, pharmacodynamic and biochemical mechanisms of anthracycline cardiotoxicity and correlate them to clinical phenotypes of cardiac dysfunction. Attention is paid to bioactivation mechanisms that converted anthracyclines to reactive oxygen species (ROS) or long lived secondary alcohol metabolites. Preclinical aspects and clinical implications of the "oxidative stress" or "secondary alcohol metabolite" hypotheses are discussed on the basis of literature that cuts across bench and evidence-based medicine. Interactions of anthracyclines with comorbidities or unfavorable lifestyle choices were identified as important cofactors of the lifetime risk of cardiotoxicity and as possible targets of preventative strategies. EXPERT OPINION: Anthracycline cardiotoxicity is a multifactorial process that needs to be incorporated in a translational framework, where individual genetic background, comorbidities, lifestyles and other drugs play an equally important role. Fears for cardiotoxicity should not discourage from using anthracyclines in many oncologic settings. Cardioprotective strategies are available and should be used more pragmatically in routine clinical practice. PMID- 21635150 TI - Novel ways to target brain tumour metabolism. AB - INTRODUCTION: Glioblastoma remains a highly aggressive primary brain cancer with very poor prognosis. The detection of mutations in the metabolic enzyme isocitrate dehydrogenase in gliomas, has broadened our view of tumourigenic mechanisms. Together with renewed awareness of tumour-specific energy metabolism, research is pointed towards novel ways for targeting brain cancer. AREAS COVERED: This paper reviews recent knowledge on the possible tumourigenic mechanism of mutant isocitrate dehydrogenase, and provides a detailed overview of cancer specific metabolic enzymes associated with glycolysis and intracellular pH regulation. It also discusses available drugs that may serve as a basis for novel drug development to target metabolic transformation in gliomas. EXPERT OPINION: Despite the fact that energy metabolism is a very basic cellular process, tumour specific alterations in key metabolic processes represent promising targets for glioma treatment. Novel therapies against gliomas, including those that target metabolic transformation, need to consider the genetic background of the individual tumours, to allow the correlation of treatment response with the underlying biological status, both in preclinical and clinical studies. PMID- 21635151 TI - Biology and novel treatment options for XLA, the most common monogenetic immunodeficiency in man. AB - INTRODUCTION: X-linked agammaglobulinemia (XLA) is the most common primary immunodeficiency in man, and is caused by a single genetic defect. Inactivating mutations in the Bruton's tyrosine kinase (BTK) gene are invariably the cause of XLA,. XLA is characterized by a differentiation arrest at the pre-B cell stage, the absence of immunoglobulins and recurrent bacterial infections, making it an insidious disease that gradually disables the patient, and can result in death due to chronic lung disease. Current treatment involves prophylactic antibiotics and immunoglobulin infusions, which are non-curative. This disease is a good candidate for curative hematopoietic stem cell (HSC)-based gene therapy, which could correct the B cell and myeloid deficiencies. AREAS COVERED: This paper reviews the basic biology of BTK in B cell development, the clinical features of XLA, and the possibilities of gene therapy for XLA, covering the literature from 1995 to 2010. EXPERT OPINION: Work from various laboratories demonstrates the feasibility of using gene-corrected HSCs to complement the immune defects of Btk deficiency in mice. We propose that it is timely to start clinical programs to develop stem cell based therapy for XLA, using gene-corrected autologous HSC. PMID- 21635153 TI - Plasma protein binding and blood-free concentrations: which studies are needed to develop a drug? AB - INTRODUCTION: The plasma protein binding of drugs and metabolites is known to influence their pharmacokinetics and, therefore, their effects. Evaluating the extent and the linearity of protein binding is an essential piece of information that has to be generated during drug development. Blood cell partitioning has a similar relevance. AREAS COVERED: This paper summarizes the regulatory requirements and focuses particularly on two questions pertaining to the drug development process. The first of these questions asks when is it necessary to perform detailed clinical studies on protein binding while the second asks whether the in vitro studies presently performed in plasma produce biased information. EXPERT OPINION: The authors propose that clinical ex vivo protein binding studies should be performed on highly bound compounds (a definition of highly bound is suggested as > 95%). They also propose that in vitro studies, to measure the free drug, should be performed in whole blood, rather than in plasma, particularly if binding to proteins or blood cells is nonlinear. PMID- 21635152 TI - Inhibition of Akt with small molecules and biologics: historical perspective and current status of the patent landscape. AB - INTRODUCTION: Akt plays a pivotal role in cell survival and proliferation through a number of downstream effectors; unregulated activation of the PI3K/PTEN/Akt pathway is a prominent feature of many human cancers. Akt is considered an attractive target for cancer therapy by the inhibition of Akt alone or in combination with standard cancer chemotherapeutics. Both preclinical animal studies and clinical trials in humans have validated Akt as an important target of cancer drug discovery. AREA COVERED: A historical perspective of Akt inhibitors, including PI analogs, ATP-competitive and allosteric Akt inhibitors, along with other inhibitory mechanisms are reviewed in this paper with a focus on issued patents, patent applications and a summary of clinical trial updates since the last review in 2007. EXPERT OPINION: A vast diversity of inhibitors of Akt, both small molecule and biologic, have been developed in the past 5 years, with over a dozen in various phases of clinical development, and several displaying efficacy in humans. While it is not yet clear which mechanism of Akt inhibition will be optimal in humans, or which Akt isoforms to inhibit, or whether a small molecule or biologic agent will be best, data to all of these points will be available in the near future. PMID- 21635154 TI - Pharmacokinetics and pharmacodynamics of calcium-vitamin D(3) chewable tablets: a single-blind, multiple-dose study in postmenopausal women. AB - OBJECTIVE: The objective of this study is to investigate the effect of a newly developed calcium carbonate-vitamin D3 chewable tablet formulation (600 mg of calcium + 400 IU of vitamin D3) on serum/urine calcium and serum parathyroid hormone (PTH) as measures of intestinal calcium absorption compared to a placebo. METHODS: This is a subject-blind, sequential study in 24 healthy postmenopausal women. Each subject received two placebo tablets once daily for 3 days (days -3 to -1) immediately followed by two calcium-vitamin D3 tablets (test) during the subsequent 3 days (days 1-3). Serial blood sampling and 24-h urine collection took place on days -1 and 3. The subjects fasted until 6 h post-dosing. Total urinary calcium excretion (Ae(0-24 h)) and AUC(0-6 h) for serum calcium were the primary outcome variables and were compared between treatments using a paired sample t-test. RESULTS: Ae(0-24 h) increased by 42% (uncorrected, p = 0.0001) and 30% (creatinine-corrected, p = 0.0001), after intake, compared with the placebo; serum calcium exposure (AUC(0-6 h)) was also, statistically, significantly greater. PTH, in serum, decreased by 28% (AUC(0-6 h), p = 0.0001) and 14% (AUC(0 24 h), p = 0.0009) when compared with the placebo. CONCLUSION: Daily intake of 1200 mg of calcium and 800 IU of vitamin D3, with a new chewable tablet, resulted in increased intestinal calcium absorption compared to the results from the placebo as confirmed by direct and indirect pharmacokinetic/pharmacodynamic measures. PMID- 21635155 TI - A survey of small molecule glucagon receptor antagonists from recent patents (2006 - 2010). AB - INTRODUCTION: The ever increasing prevalence of type 2 diabetes mellitus (T2DM) in the developed and developing nations calls for the introduction of new and more effective treatments. Glucagon receptor (GCGR) antagonists are highly validated in preclinical models of T2DM and thus have the potential to be developed as a new therapy. Small molecule GCGR antagonists have been an active area of research since the 1990s. As evidenced from the number of patents and laboratories involved, these efforts have accelerated during the last decade. AREAS COVERED: During the period 2006 - 2010, there were numerous patent publications from several laboratories claiming the discovery of novel small molecule GCGR antagonists. Herein, we present our interpretation of these new patent publications as well as follow-up disclosures appearing in the peer reviewed literature. This paper provides an up-to-date overview of the field of small molecule GCGR antagonism as a potential treatment for T2DM. Attempts were made wherever possible to identify preferred or representative compounds from the patent applications reviewed. In vitro and in vivo data are also discussed where they were disclosed. EXPERT OPINION: The novel small molecule GCGR antagonists reviewed here represent many diverse structural motifs. Some molecules are very potent antagonists of the GCGR in in vitro assays with acceptable selectivity. Some have intriguing in vivo activity in models of T2DM in a variety of preclinical species. It is to be hoped that clinical developments following these preclinical discoveries might result in a long-awaited new treatment for T2DM. PMID- 21635160 TI - Calcium supplements--friend or foe? PMID- 21635161 TI - Mapping QTL for grain yield, yield components, and spike features in a doubled haploid population of bread wheat. AB - A doubled haploid (DH) population derived from a cross between the Japanese cultivar 'Fukuho-kumogi' and the Israeli wheat line 'Oligoculm' was used to map genome regions involved in the expression of grain yield, yield components, and spike features in wheat (Triticum aestivum L). A total of 371 markers (RAPD, SSR, RFLP, AFLP, and two morphological traits) were used to construct the linkage map that covered 4190 cM of wheat genome including 28 linkage groups. The results of composite interval mapping for all studied traits showed that some of the quantitative trait loci (QTL) were stable over experiments conducted in 2004 and 2005. The major QTL located in the Hair-Xpsp2999 interval on chromosome 1A controlled the expression of grains/spike (R(2) = 12.9% in 2004 and 22.4% in 2005), grain weight/spike (R(2) = 21.4% in 2004 and 15.8% in 2005), and spike number (R(2) = 15.6% in 2004 and 5.4% in 2005). The QTL for grain yield located on chromosomes 6A, 6B, and 6D totally accounted for 27.2% and 31.7% of total variation in this trait in 2004 and 2005, respectively. Alleles inherited from 'Oligoculm' increased the length of spikes and had decreasing effects on spike number. According to the data obtained in 2005, locus Xgwm261 was associated with a highly significant spike length QTL (R(2) = 42.33%) and also the major QTL for spikelet compactness (R(2) = 26.1%). PMID- 21635163 TI - The management of patients with thyroid eye disease after bilateral orbital 3 wall decompression. AB - INTRODUCTION: Thyroid eye disease is the most common cause of unilateral and bilateral proptosis in adults. Orbital decompression surgery may cause and/or worsen a pre-existing ocular motility disorder. METHODS: A retrospective review was carried out of all bilateral 3 wall orbital decompressions for severe thyroid eye disease performed between January 2002 and December 2004 by one surgeon. Subsequent surgeries were recorded. RESULTS: Seventy-four patients were identified, 59 (80%) females and 15 (20%) males. Mean age at the time of decompression was 46 years. Fifteen (20%) patients complained of diplopia due to strabismus prior to decompression surgery and 20 (27%) developed new diplopia postsurgery. Twenty patients (27%) required no further intervention following decompression surgery; the remainder underwent an average of 2.5 procedures. Strabismus surgery was performed in 32 (43%) patients. The mean time from the decompression to first strabismus surgery was 12 months. Forty-three (58%) patients underwent lid surgery. The mean time from decompression to first lid surgery was 16 months. CONCLUSION: This study demonstrates how this group of complex patients required multiple surgical procedures within an extended timescale, therefore requiring several in- and outpatient visits. PMID- 21635164 TI - Role of inferior rectus botulinum toxin injection in iatrogenic vertical strabismus. AB - INTRODUCTION: We describe the uses and effectiveness of botulinum toxin to the inferior rectus muscle in iatrogenic vertical strabismus. METHODS: A retrospective review of our database to identify patients who had undergone inferior rectus botulinum toxin injection between 1982 and 2006 for iatrogenic vertical strabismus. RESULTS: There were 13 patients, 5 female, with an age range of 14 to 78 years. Eight left eyes were injected. The mean period of follow-up was 3.3 years. Etiologies included post-local anesthetic, post-plaque radiotherapy for malignant melanoma, adherence syndrome after inferior oblique surgery, and post-botulinum toxin to the levator muscle to produce a temporary therapeutic ptosis. Reversal on first injection occurred in 7 patients and occurred in a further 2 patients on second injection. Pre-injection binocularity was not demonstrable in 3 patients; 2 of these patients did have binocular vision demonstrable after botulinum toxin injection. Seven patients had one injection, 3 patients had two, 1 had three, 1 had five, and 1 had six. DISCUSSION: The data demonstrate the range of indications that can be successfully treated and managed with inferior rectus botulinum toxin injection in iatrogenic vertical strabismus. PMID- 21635165 TI - Comparison of cyclodeviation and duction measurement in Graves' orbitopathy patients using different devices. AB - PURPOSE: To compare measurement outcomes of different devices measuring cyclodeviation and ductions in Graves' orbitopathy (GO) patients. METHODS AND MATERIALS: Cyclodeviation in GO patients was measured using the Harms tangent screen (HTS), the cycloforometer of Franceschetti, and the synoptometer. Ductions were measured using the modified perimeter, the Goldmann perimeter and the Maddox tangent scale (MTS). RESULTS: In 13 patients, cyclodeviation in primary position, up-, and downgaze was measured with the above-mentioned devices. The mean differences ranged from 0.3o to 3.1o and were smallest between the HTS and the cycloforometer (89% of all measurements within 2o difference). Measurement of abduction, adduction, elevation, and depression using the modified perimeter, Goldmann perimeter, and MTS were obtained in another 13 patients. The mean differences ranged from 1.2o to 12.9o and were smallest between the modified perimeter and the Goldmann perimeter (92% of all measurements <= 8o). CONCLUSIONS: The HTS and cycloforometer produce interchangeable measurement outcomes. The modified perimeter and the Goldmann perimeter are interchangeable as well. However, the synoptometer and the MTS are not suitable for comparative analysis. PMID- 21635166 TI - The response AC/A ratio: differences between inducing and relaxing accommodation at different distances of fixation. AB - PURPOSE: To determine if there is a difference between the response AC/A ratios when measured using the gradient method at near and distance fixation with plus and minus lenses respectively in young adults with normal binocular single vision. METHODS: A repeated measures design was used. The accommodative response of the right eye was measured objectively using the Shin-Nippon SRW-5000 autorefractor (Grand Seiko Company, Fukuyama, Japan) (open view) at 33 cm with and without plus lenses (2DS or 3DS) and at 3.8 m with and without minus lenses (2DS or 3DS) dependent on the participants' ability to obtain subjectively "clear" vision. The angle of deviation was measured using the alternate prism cover test at 33 cm and 3.8m fixing with the right eye with the participant sat at the autorefractor. LogMAR 0.0 (6/6) was used for fixation. Response AC/A ratios were calculated. RESULTS: Twenty-five participants were examined; mean and standard deviation of their ages were 21.2 +/- 4.04 years. The mean and standard deviation of the near response AC/A ratios was 4.73 +/- 2.34/1 and at distance was 3.05 +/- 1.71/1. Pearson's Product Moment Correlation Coefficient showed no correlation between the 2 sets of data. Paired t-test showed that there was a statistically significant difference between the near and distance response AC/A ratios (t = 3.30, p = 0.003). The difference was found to be greater in participants who were non-orthoptic students. CONCLUSION: The response AC/A ratio was found to be slightly higher at 33 cm with plus lenses than at 3.8 m with minus lenses. No reason was identified for this difference but adaptation and perceptual effects could be further explored. PMID- 21635167 TI - Inferior oblique recession: an efficient technique. AB - INTRODUCTION: We describe a novel, simple, rapid technique for inferior oblique recession. METHOD: Access to the muscle is gained through a peripheral radial conjunctival incision midway between the lateral and inferior recti. The anterior edge of the inferior oblique muscle is identified coursing parallel to the limbus on the undersurface of Tenon's capsule, at the apex of the exposure. A locking vicryl suture is placed 1 mm from the muscle insertion. The insertion is cut along the globe under direct vision and attached to the sclera 2 mm posterior and lateral to the lateral border of inferior rectus. The conjunctiva is closed with interrupted vicryl sutures. RESULTS: The technique described allows direct visualization of the muscle throughout the procedure. In addition, bleeding is usually avoided and tissue manipulation is minimized, thus ensuring good visibility throughout the procedure and minimizing the risk of scarring. CONCLUSION: Severence of the inferior oblique muscle insertion under direct vision, facilitated by good traction of the globe in the same meridian, affords a rapid minimally invasive approach to inferior oblique recession. PMID- 21635168 TI - Tissue adhesives as an alternative for conjunctival closure in strabismus surgeries. AB - PURPOSE: Comparison of conjunctival closure with fibrin glue and conventional closure in strabismus surgeries. METHODS: Twenty-nine patients undergoing strabismus surgery were studied. Group 1 included twelve cases in whom Tisseel was used for closure and group 2 included seventeen cases in whom 7/0 Vicryl was used. Severity of redness, watering, pain, and discomfort in opening eyes were evaluated postoperatively. RESULTS: In the first hour, severe pain was experienced in both groups without statistical significance; but in group 1, pain was less severe in the 12th and 24th hour (P < .0001). After the first hour severity of watering was less in group 1 (P < .0001). Discomfort in opening eyes was detected in fifteen patients in group 2 in the first 12 hours where only one case with fibrin glue had discomfort. In the second week, no differences in severity of complaints were seen. CONCLUSIONS: Conjunctival closure with Tisseel following strabismus surgery might be an alternative to suturing. PMID- 21635169 TI - A survey of the management of globe perforation during strabismus surgery in the United Kingdom. AB - INTRODUCTION: Globe perforation is a potentially devastating complication of strabismus surgery. The purpose of this study is to investigate the management of suspected globe perforation encountered during strabismus surgery in the United Kingdom. METHODS: A postal survey of all consultant ophthalmologists in the United Kingdom was undertaken. The questionnaire solicited data on the management and outcomes of globe perforation encountered during strabismus surgery. RESULTS: Five hundred and seventeen of 850 (60.8%) surgeons responded to the survey. Of these, 240 (46.4%) performed strabismus surgery, of whom 58.8% performed fewer than 50, 32.1% performed 50 to 100, and 8.3% performed more than 100 surgical procedures annually. Ninety-eight (40.8%) had suspected globe perforation during strabismus surgery, of whom 86 subsequently carried out dilated fundoscopy. There were 74 reported globe perforations. Fifty-four detailed treatment with cryopexy (50%), retinal laser (31.5%), both (9.3%), cautery (1.9%), and not reported (7.4%). In 27% of cases surgeons chose not to treat the perforation observed postoperatively. Thirty-five respondents prescribed additional antibiotics. Visual loss occurred in 6 cases, with 2 cases of endophthalmitis, 2 retinal detachments, 1 suprachoroidal hemorrhage, and 1 from a chorioretinal scar. DISCUSSION: Complications leading to visual loss were infrequent. Of the surgeons reporting globe perforations, approximately three quarters chose to treat the injury and one quarter chose to observe without treatment. Cryotherapy was the most popular intervention. Further prospective study is required to elucidate optimal management strategies. PMID- 21635170 TI - Dissociated vertical divergence: cortical or subcortical in origin? PMID- 21635172 TI - Philippe de la Hire's description of the mechanism leading to strabismus in 1694. PMID- 21635173 TI - Dissertation on various accidents of sight. PMID- 21635174 TI - Improving the selective cancer killing ability of ZnO nanoparticles using Fe doping. AB - This work reports a new method to improve our recent demonstration of zinc oxide (ZnO) nanoparticles (NPs) selectively killing certain human cancer cells, achieved by incorporating Fe ions into the NPs. Thoroughly characterized cationic ZnO NPs (~6 nm) doped with Fe ions (Zn(1-x )Fe (x) O, x = 0-0.15) were used in this work, applied at a concentration of 24 MUg/ml. Cytotoxicity studies using flow cytometry on Jurkat leukemic cancer cells show cell viability drops from about 43% for undoped ZnO NPs to 15% for ZnO NPs doped with 7.5% Fe. However, the trend reverses and cell viability increases with higher Fe concentrations. The non-immortalized human T cells are markedly more resistant to Fe-doped ZnO NPs than cancerous T cells, confirming that Fe-doped samples still maintain selective toxicity to cancer cells. Pure iron oxide samples displayed no appreciable toxicity. Reactive oxygen species generated with NP introduction to cells increased with increasing Fe up to 7.5% and decreased for >7.5% doping. PMID- 21635176 TI - Strain response of an instrumented intramedullary nail to three-point bending. AB - OBJECTIVES: An experimental biomechanical evaluation of an instrumented intramedullary nail (TriGen(r) META Nail, Smith&Nephew(r)) was undertaken. The objectives were two-fold. The first was to identify the most sensitive strain gauge positions and orientations on the nail, and the second was to demonstrate that the nail was capable of detecting changes in stiffness of the nail-bone composite. The function of the instrumented nail is to quantify fracture healing objectively and directly, and so to predict delayed repair or non-union 2 months before current methods. METHODS: Eight flat pockets were machined onto the surface of the nail and three strain gauges attached in each pocket. The instrumented nail was inserted into fourth generation biomechanical grade Sawbones(r) tibiae with three different fracture configurations as well as into a non-fractured bone. The nail-bone composite was loaded in three-point bending at five positions to determine the strain changes in each of the eight strain gauge pockets located along the length of the nail. To simulate callus in the simplest way and to increase the stiffness of the nail-bone composite, loops of duct tape in multiples of four were applied over the fracture locus. A three-point loading jig was used to obtain the change in strain with increasing stiffness. Relative displacement of the bone ends was quantified using radiostereometric analysis. RESULTS: There was no single position of greatest strain sensitivity for all fracture types. The greatest change in strain occurred when the strain gauge pocket and fracture line were closest. Applying the loading moment directly over the strain gauge pocket also maximised its sensitivity. The duct tape callus simulation showed that the instrumented nail was able to detect a change in stiffness of less than 4.1 Nm/ degrees . CONCLUSIONS: It has been shown that the instrumented nail can detect physiologically relevant changes in stiffness, and so to provide a useful function as an objective monitor of fracture repair. PMID- 21635175 TI - Ketogenic diet prevents alterations in brain metabolism in young but not adult rats after traumatic brain injury. AB - Previous studies have shown that the change of cerebral metabolic rate of glucose (CMRglc) in response to traumatic brain injury (TBI) is different in young (PND35) and adult rats (PND70), and that prolonged ketogenic diet treatment results in histological and behavioral neuroprotection only in younger rat brains. However, the mechanism(s) through which ketones act in the injured brain and the biochemical markers of their action remain unknown. Therefore, the current study was initiated to: 1) determine the effect of injury on the neurochemical profile in PND35 compared to PND70 rats; and 2) test the effect of early post-injury administration of ketogenic diet on brain metabolism in PND35 versus PND70 rats. The data show that alterations in energy metabolites, amino acid, and membrane metabolites were not evident in PND35 rats on standard diet until 24 h after injury, when the concentration of most metabolites was reduced from sham-injured values. In contrast, acute, but transient deficits in energy metabolism were measured at 6 h in PND70 rats, together with deficits in N acetylaspartate that endured until 24 h. Administration of a ketogenic diet resulted in significant increases in plasma beta-hydroxybutyrate (betaOHB) levels. Similarly, brain betaOHB levels were significantly elevated in all injured rats, but were elevated by 43% more in PND35 rats compared to PND70 rats. As a result, ATP, creatine, and phosphocreatine levels at 24 h after injury were significantly improved in the ketogenic PND35 rats, but not in the PND70 group. The improvement in energy metabolism in the PND35 brains was accompanied by the recovery of NAA and reduction of lactate levels, as well as amelioration of the deficits of other amino acids and membrane metabolites. These results indicate that the PND35 brains are more resistant to the injury, indicated by a delayed deficit in energy metabolism. Moreover, the younger brains revert to ketones metabolism more quickly than do the adult brains, resulting in better neurochemical and cerebral metabolic recovery after injury. PMID- 21635177 TI - National Institute for Health and Clinical Excellence: the Medical Technologies Evaluation Programme. AB - The Medical Technologies Evaluation Programme at NICE focuses on the evaluation of innovative medical technologies, including devices and diagnostics. It's designed to enable new medical technologies, or important modifications of existing ones, to be used more quickly and consistently in the NHS. This programme will both complement and operate in conjunction with NICE's existing technology appraisal capacity, which continues to evaluate new pharmaceutical and biotechnology products. The direct website link is: http://www.nice.org.uk/MT PMID- 21635179 TI - Reduced in vivo cytotoxicity and increased Mycobacterial burden are associated with virulent Mycobacterium tuberculosis strains during lung infection. AB - Cytotoxic cellular responses are crucial for clearing intracellular pathogens and generating host resistance. Experimental pulmonary tuberculosis is associated with an early delay in T cell responses and with elevated lung bacterial burden during chronic infection. In this study we quantified the in vivo cytotoxicity and the mycobacterial burden from two pertinent tissues in groups of mice infected each with a mycobacterial strain of different virulence. None of the strains induced cytotoxic responses during early (day 14) infection. Interestingly, at 21 and 60 days post-infection, Mycobacterium canettii (lowest virulence) triggered the strongest in vivo cytotoxicity both in lungs and mediastinal lymph nodes. In contrast, Mycobacterium tuberculosis H37Rv (intermediate virulence) and Beijing strains (highest virulence) induced lower cytotoxic responses, and exhibited high bacterial growth, especially in lungs. These in vivo data suggest that virulence of Mycobacterium strains are somehow associated with subverting cytotoxic responses, thus contributing to early bacterial replication and subsequent persistence in the lungs. PMID- 21635180 TI - Women's attitudes regarding mode of delivery and cesarean delivery on maternal request. AB - OBJECTIVE: To explore women's attitudes and beliefs regarding cesarean delivery and cesarean delivery on maternal request (CDMR). STUDY DESIGN: Anonymous questionnaires assessing patient demographics, knowledge, and attitudes about CDMR were distributed at the time of routine mid-trimester ultrasound appointment. RESULTS: Eight hundred thirty three out of 3929 (21.2%) potential participants completed the questionnaire. About 81.7% of participants indicated that they believed that vaginal delivery was a safer alternative for the mother and 72.8% believed that it was safer for the fetus. While only 6.1% of women thought that CDMR was 'a good idea', most believed that women should have the right to choose their mode of delivery and that the option should be offered to everyone (85.9% and 79.6%, respectively). Socioeconomic and demographic variables did not significantly influence the participants' responses. CONCLUSION: Majority of women believe that vaginal delivery is safer for the mother and baby and would prefer to have a vaginal delivery if given the option. PMID- 21635181 TI - Ingroup identity as an obstacle to effective multiprofessional and interprofessional teamwork: findings from an ethnographic study of healthcare assistants in dementia care. AB - Rising dementia incidence is likely to increase pressures on healthcare services, making effective well coordinated care imperative. Yet, barriers to this care approach exist which, we argue, might be understood by focussing on identity dynamics at the frontlines of care. In this article, we draw upon findings from an ethnographic study of healthcare assistants (HCAs) from three dementia wards across one National Health Service mental health trust. Data revealed that the HCAs are a close-knit 'in-group' who share low group status and norms and, often highlight their own expertise in order to promote self worth. HCAs' social identity is considered as a barrier to effective teamwork with strong ingroup behaviour suggested as a consequence of their marginalisation. We explore these findings with reference to social identity theory (Tajfel, 1974; Turner, 1978 ) and discuss implications for delivering multiprofessional and interprofessional care. PMID- 21635182 TI - Preparation for becoming members of health care teams: findings from a 5-year evaluation of a student interprofessional training ward. AB - SETTING: An orthopaedic interprofessional training ward manned by students at a University Hospital. OBJECTIVE: To assess to what extent students from different undergraduate programmes evaluated the effects of a 2-week rotation at the ward on their professional roles and the value of teamwork within health care. METHOD: A questionnaire was filled in by 841 students by the end of a 2-week rotation at the ward over 5 years. Questions concerned students' estimate of how the rotation had strengthened their insight into their own future professional role, into other students' professional roles and into the value of teamwork within health care. Differences between the different student categories and possible associations between the different estimates were statistically analysed. RESULTS: Students from all programmes reported that the rotation at the student ward had dramatically strengthened their insight about their own future professional role as well as the roles of the other professions and the value of teamwork within health care. CONCLUSION: Health care is a complicated system based on cooperation and professionalism. We suggest that interprofessional training of students from all professions within health care should be a part of their education as a worthwhile preparation for their future professional work and to ensure high-quality health care. PMID- 21635183 TI - Peeling the layers: a grounded theory of interprofessional co-learning with residents of a homeless shelter. AB - Clients, patients, families, and communities must be conceived as partners in care delivery, not just as recipients (D'Amour, D. & Oandasan, I. (2005). Journal of Interprofessional Care, 19(Suppl.), 8-20). Health-care students need an opportunity to understand community member self-determination, partnership, and empowerment (Scheyett, A., & Diehl, M. ( 2004 ). Social Work Education, 23(4), 435-450), within the frame of interprofessional education (IPE) where community members are involved as teachers and learners. The aim of this grounded theory research was to determine the conditions that support health-care students to learn with, from, and about community members. This study took place in a shelter for the homeless where nursing and social work students learned interprofessionally along with residents and clients of the shelter. Data were gathered through 7 months of participant observation, interviews, and focus groups. The interprofessional co-learning theory that emerged introduces the three phases of entering, engaging, and emerging, which co-learners experienced at different levels of intensity. This article outlines the conditions that support each of these phases of the co-learning process. This interprofessional co-learning theory provides a basis for further development and evaluation of IPE programs that strive to actively include community members as teachers and learners, experts, and novices together with service providers, students, and faculty members. PMID- 21635184 TI - The "elephant in the room": using emotion management to uncover hidden discourses in interprofessional collaboration and teamwork. PMID- 21635185 TI - What is the pressure in chronic subdural hematomas? A prospective, population based study. AB - Surgery for chronic subdural hematoma (CSDH) is performed to relieve brain displacement and high intracranial pressure (ICP). However, the intraoperative impression is often that the pressure inside the CSDH is low, despite marked clinical symptoms. We wanted to quantify the CSDH pressure and relate this to radiological and clinical characteristics. This prospective, population-based study of unilateral CSDHs was conducted over a 3-year period. CSDHs that were secondary to other conditions, re-operations, or CSDHs requiring other procedures than burr hole craniostomy under local anesthesia were excluded. Subdural pressure registration was performed via a simple manometric technique, and full compliance with a standardized protocol was mandatory. Sixty patients were included (mean age 76.2 years; for men, 77.4, and for women, 72.9). The mean pressure in the CSDHs was 15.2 cm H(2)O (range, 0-40) with no gender difference. Men had significantly larger volumes (mean 158.1 vs. 103.2 cm(3)) and midline shifts (mean 1.04 vs. 0.68 cm) than did women. Large hematomas with large midline shifts had higher pressures and more often required repeat surgery. With a patient's increasing age, the volumes and midline shifts seemed to become larger, whereas the pressures became lower. We did not find an association between repeat surgery and pressure or age. Our results are generally in line with those of previous studies reporting quantitative pressure registrations. However, there are important disparities regarding methodology, not least when comparing with various subjective scales that are widely used in clinical practice. A mean subdural pressure of 15.2 cm H(2)O is probably within the range of a normal ICP. PMID- 21635186 TI - A simple, sensitive, rapid and specific detection method for Bisphenol A based on Fluorescence Polarization Immunoassay. AB - A rapid, sensitive, specific and simple method based on Fluorescence Polarization Immunoassay (FPIA) was developed for the quantitative detection of Bisphenol A in water. 4, 4-Bis (4-hydroxyphenyl) valeric acid (BHPVA) was selected as the hapten to produce the immunogen. Fluorescein-labeled Bisphenol A derivatives (tracers) with different structures were synthesized and purified by thin layer chromatography. Based on the polyclonal antibody and tracers, an optimized FPIA method was developed with a detection limit (10% inhibition) of 2 MUg L(-1) and a linear working range of 20 to 800 MUg L(-1). FPIA was suitable for screening a large number of samples. The recoveries of fortified Tai lake water samples ranged from 91.85 to 102.78%, and tap water samples ranged from 90.36 to 96.01%. The coefficients of variation were all less than 20%. This FPIA method, which did not require a complicated cleanup process, proved to be very useful for the screening of Bisphenol A in environmental water samples. PMID- 21635187 TI - Age-associated development of inflammation in Wistar rats: Effects of caloric restriction. AB - CONTEXT: Insulin resistance and type 2 Diabetes have been associated to a low grade of inflammation and their prevalence increase with ageing. OBJECTIVE: To analyse the development of inflammation in adipose tissue, liver, muscle and hypothalamus during ageing and the effects of caloric restriction. MATERIALS AND METHODS: We have analysed the expression of inflammatory cytokines (TNFalpha, IL1 beta, IL-12B and IL-6), proteins involved in macrophage recruitment (MCP-1, CCR2), TLR4 and macrophage markers (CD11c, CD11b and arginase1). Immunohistochemistry of macrophages has also been performed. RESULTS: All studied tissues present signs of inflammation during ageing, but with different pattern and intensity. Caloric restriction decreases the expression of most of inflammatory markers. DISCUSSION AND CONCLUSIONS: These data indicate a role of adiposity in the development of inflammation and insulin resistance during ageing. Dietetic intervention could be a useful tool to ameliorate the development of inflammation and insulin resistance associated with ageing. PMID- 21635190 TI - Drugs with anticholinergic properties: a current perspective on use and safety. AB - INTRODUCTION: Many commonly used drugs have primary or secondary anticholinergic effects contributing to adverse outcomes ranging from mild-to-severe to potentially lethal. Anticholinergic adverse effects frequently occur with medications prescribed with other intended mechanisms of action, including antihistamines, antidepressants, and antipsychotics. Anticholinergic drugs are also the principal treatments of clinical conditions, such as urinary incontinence, that tend to occur in the elderly. Older patients and those with mental illness are particularly vulnerable to the adverse neuropsychiatric effects of anticholinergics as they may already have cognitive impairment. AREAS COVERED: Medline and Pubmed literature searches (1966 - the present) were performed using 'anticholinergic' and 'drug safety'. Abstracts were assessed and references scanned for appropriate articles. Here, the authors i) describe the neural pathways of the cholinergic system; ii) outline the main clinical uses and adverse effects of anticholinergic agents with a focus on cognitive impairment; and iii) discuss anticholinergic safety monitoring. EXPERT OPINION: Prescribers need to be vigilant for adverse anticholinergic effects, particularly in older patients. The symptoms may range from subtle cognitive impairment to delirium and may be due to the cumulative effect of multiple medications of modest antimuscarinic activity. The Anticholinergic Drug Scale and tables listing drugs with known anticholinergic properties may help in guiding clinical decision making to reduce anticholinergic burden. PMID- 21635189 TI - Is exposure to formaldehyde in air causally associated with leukemia?--A hypothesis-based weight-of-evidence analysis. AB - Recent scientific debate has focused on the potential for inhaled formaldehyde to cause lymphohematopoietic cancers, particularly leukemias, in humans. The concern stems from certain epidemiology studies reporting an association, although particulars of endpoints and dosimetry are inconsistent across studies and several other studies show no such effects. Animal studies generally report neither hematotoxicity nor leukemia associated with formaldehyde inhalation, and hematotoxicity studies in humans are inconsistent. Formaldehyde's reactivity has been thought to preclude systemic exposure following inhalation, and its apparent inability to reach and affect the target tissues attacked by known leukemogens has, heretofore, led to skepticism regarding its potential to cause human lymphohematopoietic cancers. Recently, however, potential modes of action for formaldehyde leukemogenesis have been hypothesized, and it has been suggested that formaldehyde be identified as a known human leukemogen. In this article, we apply our hypothesis-based weight-of-evidence (HBWoE) approach to evaluate the large body of evidence regarding formaldehyde and leukemogenesis, attending to how human, animal, and mode-of-action results inform one another. We trace the logic of inference within and across all studies, and articulate how one could account for the suite of available observations under the various proposed hypotheses. Upon comparison of alternative proposals regarding what causal processes may have led to the array of observations as we see them, we conclude that the case for a causal association is weak and strains biological plausibility. Instead, apparent association between formaldehyde inhalation and leukemia in some human studies is better interpreted as due to chance or confounding. PMID- 21635191 TI - Can morphine still be considered to be the standard for treating chronic pain? A systematic review including pair-wise and network meta-analyses. AB - OBJECTIVE: For chronic pain treatment many health care authorities consider morphine to be the reference standard for strategic decisions in pain therapy. Although morphine's effectiveness is clear and its cost is low, it's unclear whether morphine should remain the first choice or reference treatment. RESEARCH DESIGN AND METHODS: We performed a systematic review to evaluate the evidence available to support the position of morphine as the reference standard for step III opioids based on efficacy and tolerability outcomes. RESULTS: The search yielded 5,675 titles and 56 studies were included. Considerable heterogeneity precluded pair-wise meta-analysis on change of pain intensity and no difference between morphine and other opioids were found for tolerability outcomes. The network meta-analysis showed no statistically significant difference in change of pain intensity between morphine and oxycodone, methadone and oxymorphone. Compared to morphine, patients using buprenorphine are more likely to discontinue treatment due to lack of effect (OR 2.32, 95% CI 1.37 to 3.95). Patients using methadone are more likely to discontinue due to adverse events (OR 3.09, 95% CI 1.14 to 8.36), whereas this risk is decreased for patients using fentanyl (OR 0.29, 95% CI 0.17 to 0.50) or buprenorphine (OR 0.30, 95% CI 0.16 to 0.53). The most important limitation of this review is that the included studies are heterogeneous with regard to study population and intervention, which may affect the pooled effect estimates. The main strength is that we only included parallel RCTs, the strongest design for intervention studies. CONCLUSIONS: The current evidence is moderate, both in respect to the number of directly comparative studies and in the quality of reporting of these studies. No clear superiority in efficacy and tolerability of morphine over other opioids was found in pair-wise and network analyses. Based on these results, a justification for the placement of morphine as the reference standard for the treatment of severe chronic pain cannot be supported. PMID- 21635192 TI - In planta reduction of maize seedling stalk lesions by the bacterial endophyte Bacillus mojavensis. AB - Maize (Zea mays L.) is susceptible to infection by Fusarium verticillioides through autoinfection and alloinfection, resulting in diseases and contamination of maize kernels with the fumonisin mycotoxins. Attempts at controlling this fungus are currently being done with biocontrol agents such as bacteria, and this includes bacterial endophytes, such as Bacillus mojavensis . In addition to producing fumonisins, which are phytotoxic and mycotoxic, F. verticillioides also produces fusaric acid, which acts both as a phytotoxin and as an antibiotic. The question now is Can B. mojavensis reduce lesion development in maize during the alloinfection process, simulated by internode injection of the fungus? Mutant strains of B. mojavensis that tolerate fusaric acid were used in a growth room study to determine the development of stalk lesions, indicative of maize seedling blight, by co-inoculations with a wild-type strain of F. verticillioides and with non-fusaric acid producing mutants of F. verticillioides. Lesions were measured on 14-day-old maize stalks consisting of treatment groups inoculated with and without mutants and wild-type strains of bacteria and fungi. The results indicate that the fusaric-acid-tolerant B. mojavensis mutant reduced stalk lesions, suggesting an in planta role for this substance as an antibiotic. Further, lesion development occurred in maize infected with F. verticillioides mutants that do not produce fusaric acid, indicating a role for other phytotoxins, such as the fumonisins. Thus, additional pathological components should be examined before strains of B. mojavensis can be identified as being effective as a biocontrol agent, particularly for the control of seedling disease of maize. PMID- 21635193 TI - Treatment of walking impairment in multiple sclerosis: an unmet need for a disease-specific disability. AB - INTRODUCTION: Walking impairment is a clinical hallmark of multiple sclerosis (MS), a chronic neurologic disease characterized by axonal demyelination and dysfunction that results in progressive disability. Until recently, there were no therapies that specifically targeted the axonal dysfunction associated with walking impairment in MS. AREAS COVERED: The purpose of this review is to discuss the unmet need for the treatment of walking impairment in MS patients and to evaluate how a new class of pharmacologic therapies, neurofunctional modifiers, potentially addresses this unmet need. Discussion is based on clinical experience and opinions supported by publications identified in the PubMed literature using the search terms 'multiple sclerosis' and 'mobility OR walking'. EXPERT OPINION: The development and approval of new treatments for MS show promise for improving adherence to therapy and increasing the potential for clinical effectiveness. Renewed emphasis on integrating strategies that target the underlying pathophysiology with those that address symptoms of concern to patients also has the potential to improve the lives of MS patients and their caregivers. The introduction of neurofunctional modifiers, such as dalfampridine for the improvement of walking impairment, may be of benefit by improving function, mobility and overall quality of life for MS patients. PMID- 21635195 TI - Anticoagulation prescription in atrial fibrillation. AB - AIMS: We seek to assess the factors associated with the anticoagulation prescription in a cohort of patients with atrial fibrillation (AF) collected from out-patient clinics. METHODS: A total of 1524 patients with a history of AF were collected from out-patients clinics. CHADS(2), CHA(2)DS(2)-VASc and HAS-BLED scores were calculated in every patient. Variables associated with anticoagulant treatment prescription were analyzed in univariant and multivariant models. RESULTS: Most patients received either anticoagulant (62%) or antiplatelet treatment (37%). Anticoagulation rates increased among higher CHADS(2) and CHA(2)DS(2)-VASc score values. A logistic regression model was performed to assess the variables associated with the prescription of anticoagulant treatment; the variables with stronger association were the presence of arrhythmia at the current visit (odds ratio (OR) 33, 95% CI 27-40, p<0.001) and lack of concomitant antiplatelet treatment (OR 0.17, 95% CI 0.14-0.21, p<0.001). CONCLUSIONS: Although prognosis of patients with AF is mainly determined by the long-term thrombotic risk, the prescription of antithrombotic therapy depends more on the bleeding risk and the immediate thrombotic risk perception. PMID- 21635196 TI - Pitfalls of clinical trials on pharmacological treatment for obstructive sleep apnoea: future directions. AB - The standard treatment for obstructive sleep apnoea (OSA) is continuous positive airway pressure (CPAP). Depending on selection criteria and quality of care, up to 50% of patients with OSA do not tolerate CPAP. For patients who cannot tolerate CPAP despite good quality care, pharmacological treatment would be a desirable alternative. The mechanisms by which pharmacological treatment is supposed to improve OSA include, amongst others, an augmentation in pharyngeal dilator muscle tone, an increase in ventilatory drive, a reduction in airway resistance and alterations in pharyngeal surface tension forces. In humans, most recent pharmacological approaches to the treatment of OSA in clinical trials have focused on modulating serotoninergic and cholinergic activities, as both have been shown to augment pharyngeal dilator muscle tone. However, currently there is not enough evidence to recommend any particular drug treatment for OSA. Methodological issues of published clinical trials on pharmacological OSA treatment make it difficult to draw definitive conclusions and inform further drug developments in this area. In this article, the pitfalls of clinical trials on pharmacological OSA therapy are summarised and potential solutions and directions for future studies are given. PMID- 21635194 TI - Identifying an indoor air exposure limit for formaldehyde considering both irritation and cancer hazards. AB - Formaldehyde is a well-studied chemical and effects from inhalation exposures have been extensively characterized in numerous controlled studies with human volunteers, including asthmatics and other sensitive individuals, which provide a rich database on exposure concentrations that can reliably produce the symptoms of sensory irritation. Although individuals can differ in their sensitivity to odor and eye irritation, the majority of authoritative reviews of the formaldehyde literature have concluded that an air concentration of 0.3 ppm will provide protection from eye irritation for virtually everyone. A weight of evidence-based formaldehyde exposure limit of 0.1 ppm (100 ppb) is recommended as an indoor air level for all individuals for odor detection and sensory irritation. It has recently been suggested by the International Agency for Research on Cancer (IARC), the National Toxicology Program (NTP), and the US Environmental Protection Agency (US EPA) that formaldehyde is causally associated with nasopharyngeal cancer (NPC) and leukemia. This has led US EPA to conclude that irritation is not the most sensitive toxic endpoint and that carcinogenicity should dictate how to establish exposure limits for formaldehyde. In this review, a number of lines of reasoning and substantial scientific evidence are described and discussed, which leads to a conclusion that neither point of contact nor systemic effects of any type, including NPC or leukemia, are causally associated with exposure to formaldehyde. This conclusion supports the view that the equivocal epidemiology studies that suggest otherwise are almost certainly flawed by identified or yet to be unidentified confounding variables. Thus, this assessment concludes that a formaldehyde indoor air limit of 0.1 ppm should protect even particularly susceptible individuals from both irritation effects and any potential cancer hazard. PMID- 21635197 TI - Ceramide formation as a target in beta-cell survival and function. AB - INTRODUCTION: Ceramide may be synthesized de novo or generated by sphingomyelinase-dependent hydrolysis of sphingomyelin. AREAS COVERED: The role of ceramide, ceramide-sensitive signaling and ion channels in beta-cell apoptosis, lipotoxicity and amyloid-induced beta-cell death. EXPERT OPINION: Ceramide participates in beta-cell dysfunction and apoptosis after exposure to TNFalpha, IL-1beta and IFN-gamma, excessive amyloid and islet amyloid polypeptide or non-esterified fatty acids (lipotoxicity). Knockout of sphingomyelin synthase 1, which converts ceramide to sphingomyelin, leads to impairment of insulin secretion. Increased ceramidase activity or pharmacological inhibition of ceramide synthetase, inhibits beta-cell apoptosis. Ceramide contributes to endoplasmatic reticulum (ER) stress, decreased mitochondrial membrane potential in insulin-secreting cells and mitochondrial release of cytochrome c into the cytosol, which are all triggers of apoptotic cell death. Ceramide-dependent signaling involves activation of extracellularly regulated kinases 1 and 2 (ERK1/2), downregulation of Period (Per)-aryl hydrocarbon receptor nuclear translocator (Arnt)-single-minded (Sim) kinase (PASK), activation of okadaic-acid sensitive protein phosphatase 2A (PP2A) and stimulation of NADPH-oxidase with generation of superoxides and lipid peroxides. Ceramide reduces the activity of voltage gated potassium (Kv)-channels in insulin-secreting cells. The role of ceramide in beta-cell survival and function may be therapeutically relevant, because ceramide formation can be suppressed by pharmacological inhibition of ceramide synthetase and/or sphingomyelinase. PMID- 21635198 TI - An update expert opinion on management and research strategies in Parkinson's disease psychosis. AB - INTRODUCTION: Psychosis, a frequent complication in Parkinson's disease (PD), contributes significantly to morbidity, mortality, nursing-home placement and quality of life. Medication side effects, issues of trial design and negative outcomes have limited clinical advances of new treatments for PD psychosis. Evidence-based medicine maintains clozapine as the most effective antipsychotic in PD without motor worsening, despite risk of agranulocytosis. Safe, effective treatments that improve psychosis without exacerbating parkinsonism are urgently needed. AREAS COVERED: This article reviews the: i) phenomenology of PD psychosis, ii) pharmacological rationale for antipsychotics in PD; iii) clinical trials of antipsychotics in PD; iv) novel research strategies such as neuroimaging, genetics and animal models; and v) associated challenges in studying and treating PD psychosis. Preparation of this review included an extensive literature search using PubMed. EXPERT OPINION: Management of PD psychosis is complex. Challenges pertaining to study design, rating scales, subject recruitment and completion have limited PD psychosis treatment trials. Novel research strategies focus on nondopaminergic systems and incorporate neuroimaging, genetic associations and animal models. These strategies also have challenges but have the potential to enhance our understanding of PD psychosis and advance the development of agents that can ultimately be tested in well designed, randomized, controlled trials. PMID- 21635199 TI - Fatigue in Parkinson's disease. AB - INTRODUCTION: Non-motor symptoms of Parkinson's disease (PD) have become increasingly recognized as central to the disease. These include somatic symptoms, such as pain and autonomic dysfunction (bladder dysfunction, constipation, dipahoresis and orthostatic hypotension) and behavioral problems, such as dementia, depression, fatigue, sleep disorders and psychosis. Research on fatigue has focused on its epidemiology with only a single report of a beneficial treatment trial, which used methylphenidate. AREAS COVERED: This review was made of all articles related to fatigue in Parkinson's disease arising in PUBMED. It will cover the types of fatigue, epidemiology, pathophysiology and treatment of fatigue in PD. EXPERT OPINION: Fatigue is a common and severe problem in Parkinson's disease. Virtually nothing is known about it aside from its epidemiology. It is an area greatly in need of investigation. PMID- 21635200 TI - Effect of early versus conventional epidural analgesia during labor on cytokine production. AB - OBJECTIVE: To compare the effect of early epidural analgesia (EEA) vs. conventional epidural analgesia (CEA) on cytokine production in mother and neonate. METHODS: Healthy parturients with uncomplicated term pregnancies were randomized into two groups: EEA - parturients who would receive epidural analgesia before onset of pain and the control group, CEA - parturients who would receive epidural analgesia after onset of pain. Cytokines were measured in maternal blood at randomization Visual Analog Scale (VAS) < 30], 24 h postpartum, and in cord blood. RESULTS: Forty-one women were studied. Epidural was performed in EEA when VAS was 23 +/- 10 and in CEA when VAS was77 +/- 10 (p < 0.0001). Background data were similar except for ruptured membranes at admission (EEA 15%, CEA 46.6%; p = 0.03), transient hypotension (EEA 20%, CEA 0%; p = 0.03), and meconium (EEA 25%, CEA 0%; p = 0.01). No significant differences were found in cytokine levels between groups at any time. Interleukin (IL)-6 levels changed significantly only in the control group (p = 0.046). There was significant correlation between baseline maternal IL-6 level and cord blood level in CEA (r = 0.59, p = 0.005), while no significant correlation existed in EEA (r = 0.33, p = 0.16). CONCLUSION: Although there was no significant difference in cytokine levels between the groups, EEA prevented the significant increase in IL-6 during labor and interrupted IL-6 fetal-maternal dependency. PMID- 21635201 TI - Posttraumatic stress symptoms following pregnancy complicated by hyperemesis gravidarum. AB - OBJECTIVE: Hyperemesis gravidarum (HG) can be accompanied by severe physical and emotional distress. Most studies have focused on the physical and psychological stress associated with this condition during the affected pregnancy. This study explores posttraumatic stress symptoms (PTSS) and negative life outcomes following HG pregnancies. METHODS: A total of 610 women (HG = 377 and control = 233) were recruited and completed an online survey. chi-square analyses were used to compare the HG and control groups on various life outcome variables. RESULTS: Eighteen percent of women with HG reported full criteria PTSS (n = 68). Negative life outcomes regarding financial and marital status, career, as well as psychological and physical well-being differed significantly for the HG groups compared to the control group (0.001 < p < 0.05). CONCLUSIONS: PTSS is common following HG pregnancies and is associated with negative life outcomes including inability to breastfeed, marital problems, financial problems, and inability of self care. PMID- 21635202 TI - Association of aneuploidy category with centrosome amplification in multiple myeloma. PMID- 21635203 TI - Classification of non-Hodgkin lymphomas in Guatemala according to the World Health Organization system. AB - The aim of this study is to report the relative frequencies of non-Hodgkin lymphoma (NHL) subtypes in Guatemala. A panel of five hematopathologists reviewed 226 consecutive biopsies and classified them according to the 2001 World Health Organization (WHO) classification. The 83 cases of diffuse large B-cell lymphoma (DLBCL) were further subclassified into germinal center B-cell-like (GCB) and non GCB subtypes. Of the 226 cases, 194 (86%) were confirmed as NHL, including 169 (87%) B-cell and 25 (13%) T- or natural killer (NK)-cell NHL. The most common subtype was DLBCL (44.3%), and the most frequent subtype among T- and NK-cell NHL was extranodal NK/T-cell lymphoma, nasal type (7.8% of all NHL). A comparison of the frequencies of NHL subtypes between Guatemala and other parts of the world showed that Guatemala is most similar to the Middle East and Asia. However, there is no significant difference in the frequency of the DLBCL subtypes compared to North America and Europe. PMID- 21635205 TI - Treatment of refractory or relapsed acquired aplastic anemia: review of established and experimental approaches. AB - In a substantial number of patients with aplastic anemia (AA), immunosuppressive therapy (IST) with antithymocyte globulin (ATG) and cyclosporine A (CSA) leads to long-lasting remissions and is thus regarded as standard therapy. However, no consensus exists on how to treat refractory or relapsed AA, especially when no related stem cell donor is available. For selected patients, matched unrelated donor stem cell transplant (MUDSCT) is an option. In addition, umbilical cord blood and haploidentical donors have been considered as an alternative source of stem cells. Patients without a suitable donor may benefit from a second cycle of ATG and CSA. Alternatives are alemtuzumab and high dose cyclophosphamide, both of which induce remission in more than 50% of patients with relapsed AA. Further experimental drugs are androgens, hematopoietic growth factors (interleukins IL 3, IL-6, and IL-11 and stem cell factor [SCF]), and the tumor necrosis factor (TNF)-targeting agent etanercept. Clinical trials with these agents are ongoing and will explore long-term outcomes and potential beneficial effects of drug combinations. PMID- 21635204 TI - Telomere length in myelodysplastic syndromes. AB - The relationship between telomere length (TL) and predisposition to myelodysplastic syndromes (MDS) remains unclear. We compared peripheral blood leukocyte (PBL) TL among cases of histologically confirmed MDS (n = 65) who were treatment-naive with no prior cancer history to age-matched controls (n = 63). Relative TL was measured in PBLs and saliva by quantitative polymerase chain reaction (PCR) and in CD15+ and CD19+ cells by flow cytometry-fluorescence in situ hybridization (flow-FISH). Human telomerase reverse transcriptase gene (hTERT) mutations were assessed by PCR. After adjustment for age and sex, relative TLs were reduced in PBLs (p = 0.02), CD15+ (p = 0.01), CD19+ (p = 0.25), and saliva (p = 0.13) in MDS cases versus controls, although only the PBL and CD15+ results were statistically significant. Among MDS cases, CD15+ and CD19+ cell TLs were positively correlated (p = 0.03). PBL TL was reduced among those occupationally exposed to paints and pesticides, but was not associated with hTERT genotype. Future studies are needed to further investigate constitutional telomere attrition as a possible predisposing factor for MDS. PMID- 21635206 TI - A case of JAK2 V617F-positive myelodysplastic/myeloproliferative neoplasm with unusual morphology, resembling acute promyelocytic leukemia-like disorder with a chronic course. PMID- 21635207 TI - Factors influencing the inhibition of protein kinases. AB - The protein kinase field is a very active research area in the pharmaceutical industry and many activities are ongoing to identify inhibitors of these proteins. The design of new chemical entities with improved pharmacological properties requires a deeper understanding of the factors that modulate inhibitor kinase interactions. In this report, we studied the effect of two of these factors--the magnesium ion cofactor and the protein substrate--on inhibitors of the type I insulin-like growth factor receptor. Our results show that the concentration of magnesium ion influences the potency of adenosine triphosphate (ATP) competitive inhibitors, suggesting an explanation for the observation that such compounds retain their nanomolar potency in cells despite the presence of millimolar levels of ATP. We also showed that the peptidic substrate affects the potency of these inhibitors in a different manner, suggesting that the influence of this substrate on compound potency should be taken into consideration during drug discovery. PMID- 21635208 TI - Exploring QSARs for inhibitory effect of a set of heterocyclic thrombin inhibitors by multilinear regression refined by artificial neural network and molecular docking simulations. AB - Several non-peptide heterocyclic compounds reported as potent thrombin inhibitors in vitro were chosen to carry out a QSAR study upon them using MLR and ANN analysis. In order to identify the best QSAR models, the input for ANN consisted of those subsets of descriptors used in the MLR models. The best QSAR models contained the SIC0 descriptor as the main topological descriptor. To identify the physical and chemical properties involved in the ligand-thrombin complexes, an automated ligand-flexible docking procedure was used. The docking results suggest that the thrombin inhibition by these heterocyclic compounds is driven by pi-pi, hydrogen bonds and salt bridge interactions. The best Gibbs free energy of ligand binding was found at the thrombin sites S1 and D. We have shown that it is possible to build MLR models with geometries taken from two different sources (semi-empirical and MD geometries) and obtain similar results. PMID- 21635209 TI - Synthesis and biological evaluation of novel 6, 7-dimethoxy-3-(4-pyridyl) 2,3,3a,4-tetrahydroindeno[1,2-c]pyrazol-2-yl-4-substituted phenylmethanone/ethanone derivatives. AB - A series of 6,7-dimethoxy-3-(4-pyridyl)-2,3,3a,4-tetrahydroindeno[1,2-c]pyrazol-2 yl-4-substituted phenylmethanone/ethanone derivatives were synthesized and in vitro activity against mycobacterium tuberculosis (MTB) and INHR-MTB were carried out. Among the synthesized compounds, compound (4h) 6,7-dimethoxy-3-(4-pyridyl) 2,3,3a,4-tetrahydroindeno[1,2-c]pyrazol-2-yl-4-pyridyl methanone was found to be the most active agent against MTB and INHR-MTB with a minimum inhibitory concentration of 0.22 MUM. PMID- 21635210 TI - Synthesis, in silico metabolic and toxicity prediction of some novel imidazolinones derivatives as potent anticonvulsant agents. AB - A series of 1,2,4-trisubstituted 5-imidazolinone derivatives were synthesized by Erlenmeyer condensation of benzoylglycine (hippuric acid) with different aldehydes in the presence of sodium acetate and acetic anhydride. The derivatives of the compounds were prepared by condensation of some known sulpha drugs with 5 oxazolone derivatives. The anticonvulsant activity of the compounds was determined by the protection of pentylenetetrazole-induced convulsions that was ranged from 10 to 60%. The compounds with p-OCH3, p-OH and o-Cl substitutions in the phenyl ring on 4(th) position of the imidazolinone ring exhibited good anticonvulsant activity. In silico metabolic and toxicity studies showed that all the compounds in the series are not likely to exhibit toxicity except the compounds IIIa, IIIb, VIa and VIb, that is predicted to show 29% mutagenicity and 53% irritation in comparison to the other compounds. The predicted lethal effect and hERG toxicity of the compounds showed that IIa, IVa, Va and Vb might be toxic at higher concentrations. The results successfully establish the synthesized imidazolinone derivatives as novel compounds with anticonvulsant properties, low predicted cardiotoxicity and lethal effects thus can be promising leads for further development as novel anticonvulsants. PMID- 21635211 TI - Synthesis and carbonic anhydrase inhibitory properties of novel bromophenols including natural products. AB - (2-Bromo-3,4-dimethoxyphenyl) (3,4-dimethoxyphenyl)methanone (10) and its derivatives with Br, one dibromide and isomeric three tribromides, were synthesized. Demethylation of these compounds afforded a series of new bromophenols. Inhibition of human cytosolic carbonic anhydrase II (hCA II) isozyme by these new bromophenols and naturally occurring 3,4,6-tribromo-5-(2,5 dibromo-3,4-dihydroxybenzyl)benzene-1,2-diol (3), and 5,5'-methylenebis(3,4,6 tribromo-benzene-1,2-diol) (4) was investigated. The synthesized compounds showed carbonic anhydrase inhibitory capacities with IC(50) values in the range of 0.7 372 MUM against hCA II. Some bromophenols investigated here showed effective hCA II inhibitory activity and might be used as leads for generating novel carbonic anhydrase inhibitors which are valuable drug candidates for the treatment of glaucoma, epilepsy, gastric and duodenal ulcers, neurological disorders, or osteoporosis. PMID- 21635212 TI - Synthesis, characterization and biological properties of thienyl derived triazole Schiff bases and their oxovanadium(IV) complexes. AB - A new series of biologically active thienyl derived triazole Schiff bases and their oxovanadium(IV) complexes have been synthesized and characterized on the basis of physical (m.p., magnetic susceptibility and conductivity), spectral (IR, 1H and 13C NMR, electronic and mass spectrometry) and microanalytical data. All the Schiff base ligands and their oxovanadium(IV) complexes have been subjected to in vitro antibacterial activity against four Gram-negative (Escherichia coli, Shigella flexneri, Pseudomonas aeruginosa, Salmonella enterica serover typhi) and two Gram-positive (Staphylococcus aureus and Bacillus subtilis) bacterial strains and, for in vitro antifungal activity against Trichophyton longifucus, Candida albican, Aspergillus flavus, Microscopum canis, Fusarium solani and Candida glabrata. Brine shrimp bioassay was also carried out to check the cytotoxic nature of these compounds. PMID- 21635213 TI - In vitro effects of some anabolic compounds on erythrocyte carbonic anhydrase I and II. AB - The in vitro effects of the anabolic compounds, zeranol, 17 beta-estradiol, diethylstilbestrol (DES), and trenbolone, on the activity of purified human carbonic anhydrase I and II were evaluated. In vitro CA enzyme activity was determined colorimetrically using the CO2 hydration method of Maren. IC50 values of the compounds that caused inhibition were determined by means of activity percentage diagrams. The IC50 concentrations of zeranol, 17 beta-estradiol, DES and trenbolone on hCA I were 94, 55, 10, 898 uM and for hCA II 89, 159, 439 and 101 MUM, respectively. PMID- 21635214 TI - The synthesis and biological activity of lipophilic derivatives of bicine conjugated with N3-(4-methoxyfumaroyl)-L-2,3-diaminopropanoic acid (FMDP)-an inhibitor of glucosamine-6-phosphate synthase. AB - A series of bis-N,N-(2-hydroxyethyl)glycine (bicine) derivatives, conjugated with an inhibitor of glucosamine-6-phosphate synthase, have been synthesized and their lipophilic and antifungal properties have been tested. The obtained compounds demonstrated higher lipophilicity than free inhibitor (FMDP) and, in consequence, an increased potential to cross the cytoplasmic membrane. All the tested compounds show better antifungal activity than parent compound. PMID- 21635215 TI - Treatment patterns and resource use among patients with comorbid diabetes mellitus and major depressive disorder. AB - OBJECTIVE: To descriptively analyze patient characteristics, treatment patterns, and medical resource use of individuals with diabetes mellitus (DM) with and without comorbid major depressive disorder (MDD). RESEARCH DESIGN AND METHODS: Using an administrative claims database, commercially insured adults diagnosed with DM were studied over the calendar years 2005 and 2006. Demographic characteristics, comorbid illnesses, medication use, resource utilization, and costs were examined descriptively among patients with DM both with and without comorbid MDD. RESULTS: Patients with DM and MDD were found to have a high resource burden, compared to patients with DM without MDD. Specifically, these patients were found to be more likely to be diagnosed with other comorbid medical conditions, to use multiple medications, and to use more healthcare services such as inpatient visits, emergency admissions, and outpatient visits. Consistent with these findings, costs for these patients were found to be $19,707 per year, compared to $11,237 for patients with DM without comorbid MDD. LIMITATIONS: The study utilizes data from an administrative claims database of insured individuals and hence, results may not be generalizable. Furthermore, the analysis is unable to examine clinical severity or indirect costs. CONCLUSION: Compared to patients with DM and no comorbid MDD, patients with DM and MDD tend to have a larger burden of disease and to use more healthcare resources. PMID- 21635216 TI - An efficient, economical slow-freezing method for large-scale human embryonic stem cell banking. AB - Human embryonic stem cells (hESCs) are one of the most interesting cell types for tissue engineering, cell therapy, basic scientific research, and drug screening. Fast advancement in these areas requires the availability of large amounts of safe and well-characterized hESCs from hESC banks. Therefore, optimized freezing protocols, allowing the cryopreservation of large amounts of hESC without direct contact with liquid nitrogen, need to be established. In this study, 6 different cryoprotector combinations [dimethylsulfoxide (DMSO), ethylene glycol, and hydroxyethylstarch (HES)] combined with 2 different application methods were screened with the VUB01 cell line, to establish a new slow-freezing protocol with high recovery rates and a good expansion capacity. Our best conditions were confirmed in 4 other hESC lines: H1, H9, 181, and UGent2. To our knowledge, this is the first time that HES is evaluated as a cryoprotector for hESCs. The use of 5% DMSO+5% HES combined with a new detachment protocol leads to efficient hESC cryopreservation. This protocol involves treating the hESC colonies with cell dissociation solution, a mild dissociation solution uncommonly used for hESC culture. A recovery ratio ranging from 45.5% to 168.2% was obtained, and these were significantly different from the other tested conditions (Student's t-test, P<0.05). The cryopreserved hESCs were morphologically comparable to control cells, exhibited a good expansion profile, were positive for pluripotent expression markers, and could still differentiate into the 3 germ layers. This new protocol allows efficient and economical hESC cryopreservation, ideal for hESC banking. PMID- 21635217 TI - Characterization of the bacterial community structure of Sydney Tar Ponds sediment. AB - The Sydney Tar Ponds is one of the largest toxic waste sites in Canada. The bacterial diversity and abundance in the Sydney Tar Ponds sediment was examined using a 16S rRNA gene clone library and denaturing gradient gel electrophoresis (DGGE) with four different primer sets. The clone library was grouped into 19 phylotypes that could be divided into five phyla: Proteobacteria (56.9%), Actinobacteria (35%), Acidobacteria (4.9%), Firmicutes (2.4%), and Verrucomicrobia (0.8%). Members of the phyla Actinobacteria (represented mainly by Mycobacterium spp.) and Alphaproteobacteria (represented by Acidocella spp.) comprised the majority of the clone library. This study also revealed that the phylogenetic results obtained from clone library analysis and from DGGE analysis, with all the primer sets, showed some variability. However, similar Mycobacterium spp. and Acidocella spp. were found in all the different DGGE analyses, again suggesting that these two genera are dominant in the Sydney Tar Ponds sediment. In addition, DGGE analysis indicated that primer sets targeting the V3 region produced results that were the most similar to those obtained with the clone library. PMID- 21635218 TI - Temporal distribution of the archaeal community in the Changjiang Estuary hypoxia area and the adjacent East China Sea as determined by denaturing gradient gel electrophoresis and multivariate analysis. AB - The archaeal community and the effects of environmental factors on microbial community distribution were investigated at five sampling sites in the Changjiang Estuary hypoxia area and the adjacent East China Sea in June, August, and October 2006. Profiles of the archaeal communities were generated by denaturing gradient gel electrophoresis of 16S rRNA genes followed by DNA sequence analysis, and the results were analyzed by multivariate statistical analysis. Denaturing gradient gel electrophoresis band patterns were analyzed by cluster analysis to assess temporal changes in the genetic diversity of the archaeal communities. Most of the October samples grouped together separately from those of June and August. Analysis of DNA sequences revealed that the dominant archaeal groups in the Changjiang Estuary hypoxia area and the adjacent East China Sea were affiliated with Euryarchaeota (mainly marine group II) and Crenarchaeota. The effects of environmental factors on the archaeal community distribution were analyzed by the ordination technique of canonical correspondence analysis. Salinity had a significant effect on the archaeal community composition. PMID- 21635219 TI - Mechanism of acid tolerance in a rhizobium strain isolated from Pueraria lobata (Willd.) Ohwi. AB - The Rhizobium sp. strain PR389 was isolated from the root nodules of Pueraria lobata (Willd.) Ohwi, which grows in acidic (pH 4.6) yellow soil of the Jinyun Mountains of Beibei, Chongqing, China. While rhizobia generally have a pH range of 6.5-7.5 for optimum growth, strain PR389 grew in a liquid yeast extract - mannitol agar medium at pH 4.6, as well as in a pH 4.1 soil suspension, suggesting acid tolerance in this specific strain of rhizobium . However, at pH 4.6, the lag phase before vigorous growth was 40 h compared with 4 h under neutral conditions (pH 7.0). For PR389, the generation time after the lag phase remained the same at different pH levels despite the different durations of the lag phase. Except in the pH 4.4 treatment, the pH of the culturing media increased from 4.6, 4.8, 5.0, and 5.5 to neutral and slightly alkaline after 70 h of culture. Chloramphenicol was added to determine if protein production was involved in the increasing pH process. Chloramphenicol significantly inhibited PR389 growth under acid stress but had little effect under neutral conditions. Proton flux measured during a short acid shock (pH 3.8) revealed that this strain has an intrinsic ability to prevent H(+) from entering cells when compared with acid-sensitive rhizobia. We propose that the mechanism for acid tolerance in PR389 involves both intracellular and extracellular processes. When the extracellular pH is lower than pH 4.4, the cell membrane blocks hydrogen from entering the cell. When the pH exceeds 4.4, the rhizobium strain has the ability to raise the extracellular pH, thereby, potentially decreasing the toxicity of aluminum in acid soil. PMID- 21635220 TI - Selection and characterization of Spiroplasma citri mutants by random transposome mutagenesis. AB - Phytopathogenic spiroplasmas can multiply in vascular plants and insects. A deeper understanding of this dual-host life could be furthered through the identification by random mutagenesis of spiroplasma genes required. The ability of the EZ::TNTM Tnp transposomeTM system to create random insertional mutations in the genome of Spiroplasma citri was evaluated. The efficiency of electroporation-mediated transformation of S. citri BR3-3X averaged 28.8 CFUs/ng transposome for 10(9) spiroplasma cells. Many transformants appearing on the selection plates were growth impaired when transferred to broth. Altering broth composition in various ways did not improve their growth. However, placing colonies into a small broth volume resulted in robust growth and successful subsequent passages of a subset of transformants. PCR using primers for the dihydrofolate reductase gene confirmed the transposon's presence in the genomes of selected transformants. Southern blot hybridization and nucleotide sequencing suggested that insertion was random within the chromosome and usually at single sites. The insertions were stable. Growth rates of all transformants were lower than that of the wild-type S. citri, but none lost the ability to adhere to a Circulifer tenellus (CT-1) cell line. The EZ::TNTM Tnp transposomeTM system represents an additional tool for genetic manipulation of the fastidious spiroplasmas. PMID- 21635222 TI - Emerging targeted therapies for pediatric acute myeloid leukemia. AB - Acute myeloid leukemia is a heterogeneous group of diseases accounting for 15-20% of all tumors diagnosed in children under 15 years of age. The past few decades have yielded remarkable improvements in long-term outcomes for children with acute myeloid leukemia. A better risk-group stratification of patients based upon clinical and biologic features, a more effective use of anti-leukemic agents and enormous improvements in supportive care have increased the probability of cure by approximately 60%. The increase in our understanding of the biology of this disease has resulted in the development of molecularly targeted therapies that are potentially more effective and less toxic than the standard approaches. We here review novel molecularly targeted drugs for the treatment of childhood acute myeloid leukemia such as monoclonal antibodies, inhibitors of signalling molecules, proteasome inhibitors and epigenetic agents. For these recently patented agents, we also provide a detailed analysis of the published preclinical data and the clinical trials that have been completed. PMID- 21635221 TI - A potential role of ruxolitinib in leukemia. AB - INTRODUCTION: An increased understanding of cellular signaling pathways, like the JAK?STAT pathway, and the identification of the JAK2 V617F mutation in the classic Philadelphia chromosome-negative myeloproliferative neoplasms (MPNs), has generated great interest in the development of targeted JAK2 inhibitors. In a recently completed Phase I?II study, ruxolitinib, a selective orally available JAK1 and JAK2 inhibitor, has shown efficacy in patients with advanced myelofibrosis. Constitutive activation of the JAK?STAT pathway has also been implicated in other hematological malignancies suggesting a potential role of JAK kinase inhibitors in these malignancies. AREAS COVERED: This article reviews the chemistry, pharmacodynamics, pharmacokinetics, clinical efficacy, safety and tolerability of ruxolitinib. The literature for this article was retrieved from PubMed database searches using the keywords ?ruxolitinib?, ?INCB 018424?, ?JAK2 inhibitors? and ?leukemia?. EXPERT OPINION: The JAK?STAT signaling pathway plays a vital role in leukemogenesis. Ruxolitinib, a potent JAK1 and JAK2 inhibitor, known to decrease spleen size and alleviate constitutional symptoms in myelofibrosis, represents a potentially promising agent for the treatment of leukemias by inhibiting the JAK?STAT signaling. Further studies of ruxolitinib, in patients with acute and chronic leukemias, are now needed to establish the clinical usefulness of this promising drug. PMID- 21635223 TI - Targeting the autolysis loop of urokinase-type plasminogen activator with conformation-specific monoclonal antibodies. AB - Tight regulation of serine proteases is essential for their physiological function, and unbalanced states of protease activity have been implicated in a variety of human diseases. One key example is the presence of uPA (urokinase-type plasminogen activator) in different human cancer types, with high levels correlating with a poor prognosis. This observation has stimulated efforts into finding new principles for intervening with uPA's activity. In the present study we characterize the so-called autolysis loop in the catalytic domain of uPA as a potential inhibitory target. This loop was found to harbour the epitopes for three conformation-specific monoclonal antibodies, two with a preference for the zymogen form pro-uPA, and one with a preference for active uPA. All three antibodies were shown to have overlapping epitopes, with three common residues being crucial for all three antibodies, demonstrating a direct link between conformational changes of the autolysis loop and the creation of a catalytically mature active site. All three antibodies are potent inhibitors of uPA activity, the two pro-uPA-specific ones by inhibiting conversion of pro-uPA to active uPA and the active uPA-specific antibody by shielding the access of plasminogen to the active site. Furthermore, using immunofluorescence, the conformation-specific antibodies mAb-112 and mAb-12E6B10 enabled us to selectively stain pro-uPA or active uPA on the surface of cultured cells. Moreover, in various independent model systems, the antibodies inhibited tumour cell invasion and dissemination, providing evidence for the feasibility of pharmaceutical intervention with serine protease activity by targeting surface loops that undergo conformational changes during zymogen activation. PMID- 21635224 TI - Ca2+-induced PARP-1 activation and ANF expression are coupled events in cardiomyocytes. AB - The nuclear protein PARP-1 [poly(ADP-ribose) polymerase-1] is activated in cardiomyocytes exposed to hypoxia causing DNA breaks. Unlike this stress-induced PARP-1 activation, our results provide evidence for Ca(2+)-induced PARP-1 activation in contracting newborn cardiomyocytes treated with growth factors and hormones that increased their contraction rate, induced intracellular Ca(2+) mobilization and its rhythmical and transient translocation into the nucleus. Furthermore, activated PARP-1 up-regulated the activity of phosphorylated ERK (extracellular-signal-regulated kinase) in the nucleus, promoting expression of the Elk1 target gene c-fos. Up-regulation of the transcription factor c-Fos/GATA 4 promoted ANF (atrial natriuretic factor) expression. Given that expression of ANF is known to be implicated in morphological changes, growth and development of cardiomyocytes, these results outline a PARP-1-dependent signal transduction mechanism that links contraction rate and Ca(2+) mobilization with the expression of genes underlying morphological changes in cardiomyocytes. PMID- 21635225 TI - Effects of epigenetic-based anti-cancer drugs in leukaemia and multiple myeloma cells. AB - Here, we focus on epigenetic changes in leukaemia and MM (multiple myeloma) cells. We show how the histone signature, DNA methylation and levels of select tumour-suppressor proteins can be affected by inhibitors of HDACs (histone deacetylases) and Dnmts (DNA methyltransferases). Both inhibitors, TSA (trichostatin A) and 5-AZA (5-azacytidine), have the ability to change the histone signature in a tumour-specific manner. In MM cells, we observed changes in H3K4 methylation, while in leukaemia cells, H3K9 methylation was especially affected by select inhibitors. Compared with normal peripheral blood lymphocytes, tumour cell samples were characterized by increased H3K9 acetylation, increased H3K4me2, H3K9me2 and HP1alpha (heterochromatin protein 1alpha) levels and specific changes were also observed for DNA methylation. Additionally, we showed that the tumour suppressor pRb1 (retinoblastoma protein) is more sensitive to epigenetic-based anti-cancer stimuli than p53. We have found significant decrease in the levels of pRb1 and p53 in both myeloma and leukaemia cells after HDAC inhibition. PMID- 21635226 TI - High frequencies of CD62L+ naive regulatory T cells in allografts are associated with a low risk of acute graft-versus-host disease following unmanipulated allogeneic haematopoietic stem cell transplantation. AB - Regulatory T cells (T(regs) ) play a key role in the prevention of acute graft versus-host disease (aGVHD). To investigate the association between T(reg) subsets and aGVHD, we prospectively analysed T cell subsets in the allografts of 35 patients undergoing myeloablative unmanipulated haematopoietic stem cell transplantation. Multivariate analysis found that patients infused with less than 0.29 * 10(6) /kg of CD4(+) CD25(high) CD45RA(+) CD62L(+) T cells during transplantation exhibited an increased incidence of II-IV aGVHD [hazard ratio (HR) = 0.000, 95% CI = 0.000-0.106, P = 0.013]. Next, we compared the reconstitution characteristics of T cell subsets between haploidentical haematopoietic stem cell transplantation (HSCT) and sibling HSCT by collecting peripheral blood samples at regular intervals (days 30, 60 and 90) after transplantation. No significant differences were observed in the reconstitution of conventional T cells between haploidentical HSCT and sibling identical HSCT. However, total counts of recovered naiveT(regs) and CD62L(+) naive T(regs) from haploidentical HSCT were significantly lower compared to sibling identical HSCT; P-values were 0.045 and 0.021, respectively. Although total counts of conventional T cells in aGVHD patients reached similar levels compared to non aGVHD patients before day 60 post-HSCT, total counts of naive T(regs) and CD62L(+) naive T(regs) in aGVHD patients did not reach similar levels to non aGVHD patients until 90 days post-HSCT. Taken together, our findings demonstrate that a large population of CD62L(+) naive T(regs) in allografts reduces the incidence of aGVHD. Further, development of aGVHD is related closely to the delayed reconstitution of the naive T(reg) population. PMID- 21635227 TI - Modulatory role of calreticulin as chaperokine for dendritic cell-based immunotherapy. AB - Heat shock proteins (HSPs) play a regulatory role for maturation of antigen presenting cells (APCs) such as dendritic cells (DCs) and macrophages. Whereas HSP70 has been shown to enhance the maturation of human DCs via a nuclear factor kappa-B (NF-kappaB)-dependent pathway, the regulatory role of calreticulin (CRT), which is a HSP with similar functions to HSP70, is not well studied. To investigate the role of CRT as adjuvant in cell activation and co-stimulatory responses we determined the effects of CRT on human APC maturation in comparison to that of HSP70. To facilitate eukaryotic endotoxin-free CRT protein expression, three different methods were compared. We demonstrate that CRT induces the maturation of human DCs and increases the production of proinflammatory cytokines via the NF-kappaB pathway. CRT-mediated maturation was qualitatively similar to that induced by HSP70. Interestingly, priming of monocytes with HSPs showed an even more prominent effect on maturation than exposure of immature DCs to these compounds. A higher expression of CD86, CD83 and CCR7 on mature DCs were found in response to CRT. Our data provide novel insights into the role of extracellular HSPs as chaperokines in the processes of APC generation and may thus be useful to improve adoptive immunotherapy. PMID- 21635228 TI - Interleukin-10 receptor expression and signalling were down-regulated in CD4+ T cells of lupus nephritis patients. AB - Studies have indicated that interleukin (IL)-10 has a pathogenic role in systemic lupus erythematosus (SLE); however, a protective effect of IL-10 in SLE was also observed. Because the exact mechanism of IL-10 signalling in the pathogenesis of SLE is unclear, this study sought to assess the expression and signalling of interleukin-10 receptor (IL-10R) in peripheral leucocytes from patients with SLE. We used flow cytometry to examine the expression of IL-10R1 on different peripheral leucocytes from 28 SLE patients, of whom 14 had lupus nephritis (LN) and 14 were healthy controls. We also examined the effects of IL-10 on phosphorylation of signal transducer and activator of transcription (STAT)-3 and STAT-1 in peripheral blood mononuclear cells (PBMCs) obtained from 13 SLE patients and seven healthy controls. Plasma cytokines were detected by flow cytometric bead array (CBA) techniques. Although IL-10R1 expression levels on each peripheral leucocyte subset from 28 SLE patients and 14 healthy controls were similar, the expression levels on CD4(+) T cells from LN patients were significantly lower than on CD4(+) T cells from controls and SLE patients without nephritis (P < 0.01). IL-10R1 expression levels on CD4(+) and CD8(+) T cells were correlated negatively with the SLE disease activity index (P < 0.01). Additionally, the phosphorylation of STAT-3 was delayed and reduced in PBMCs from LN patients and active SLE patients. Plasma IL-10 levels were significantly higher in LN patients than controls. IL-10R1 expression on CD4(+) T cells and signalling in PBMCs were down-regulated in LN patients, indicating that IL-10 and its receptor may have a special role in LN pathogenesis. PMID- 21635229 TI - Airway and interstitial lung disease are distinct entities in paediatric common variable immunodeficiency. AB - Common variable immunodeficiency (CVID) is a common primary immune deficiency, caused by undefined defects in lymphocyte function, and is treated routinely by immunoglobulin substitution. CVID complications include airway disease (AD) and interstitial lung disease (ILD). It was not known if AD and ILD in CVID have a common immunological aetiology and should be considered separate features of the same disease, or as distinct syndromes that require specialized monitoring and treatment. We used high-resolution computed tomography (CT) to diagnose AD or ILD in paediatric CVID patients. Spirometry and body plethysmography did not differentiate between ILD and AD. Patients with AD (n = 11, 20%) developed more pneumonias while children with ILD (n = 8, 15%) showed immune dysregulation characterized by autoimmune complications, more severe memory B cell reduction and expansion of non-naive cytotoxic T cells. In conclusion, ILD and AD in CVID have dissimilar clinical and immunological characteristics, suggesting distinct aetiology requiring tailored monitoring and treatment of these patient subgroups. PMID- 21635230 TI - Use of intrathecal baclofen therapy in ambulant children and adolescents with spasticity and dystonia of cerebral origin: a systematic review. AB - AIM: Studies on the use of intrathecal baclofen (ITB) for ambulant adults with spasticity and/or dystonia of cerebral origin are scarce, and are even more limited for children and adolescents. This systematic review investigates the use of ITB to improve walking, transfer ability, and gross motor activities in ambulant children and adolescents with spasticity and/or dystonia of cerebral origin. METHOD: Electronic databases (MEDLINE, CINAHL, PsycINFO, EMBASE, full Cochrane Library, and PEDro) were searched from the earliest date available until March 2011 using combined subject headings and free text if supported by the databases. Studies were included if they had examined individuals who: (1) received ITB therapy by any method (bolus injection, an external delivery system, or an implanted pump); (2) had spasticity and/or dystonia of cerebral origin; (3) were able to ambulate with or without a walking device, i.e. individuals with cerebral palsy (CP) who were in levels I to III of the Gross Motor Function Classification System or individuals with similar functional mobility if they did not have CP; and (4) were aged 18 years or under. Publications in English in peer reviewed journals reporting any type of research design, except reviews and expert opinions, were included. Studies were excluded if participants had spasticity and/or dystonia of spinal origin and if baclofen was administered only orally. Studies that compared ITB with other interventions such as botulinum toxin were also excluded. RESULTS: Two independent reviewers scored 16 studies against the guidelines for developing systematic reviews from the American Academy of Cerebral Palsy and Developmental Medicine (AACPDM). INTERPRETATION: Fifteen studies were of levels IV or V evidence and only one of level II according to the evidence levels of the AACPDM guidelines, but all were of low quality. No study was found on the use of ITB in ambulant children or adolescents with dystonia of cerebral origin. Not all studies used objective outcome measures to assess the ambulation, transfer ability, and gross motor activities of the participants. A proportion of participants showed improvement in all these areas but adverse events were common. A proportion of participants compromised their ambulatory and transfer abilities after ITB. There was no evidence to support the clinical use of ITB in ambulant individuals with hypertonicity without further rigorous longitudinal studies. PMID- 21635231 TI - Stereologic estimation of hippocampal GluR2/3- and calretinin-immunoreactive hilar neurons (presumptive mossy cells) in two mouse models of temporal lobe epilepsy. AB - PURPOSE: Hippocampal mossy cells receive dense innervation from dentate granule cells and, in turn, mossy cells innervate both granule cells and interneurons. Mossy cell loss is thought to trigger granule cell mossy fiber sprouting, which may affect granule cell excitability. The aim of this study was to quantify mossy cell loss in two animal models of temporal lobe epilepsy, and determine whether there exists a relationship between mossy cell loss, mossy fiber sprouting, and granule cell dispersion. METHODS: Representative hippocampal sections from p35 knockout mice and mice with unilateral intrahippocampal kainate injection were immunolabeled for GluR2/3, two subunits of the amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA) receptor and calretinin to identify mossy cells. Mossy fibers were immunostained against synaptoporin. KEY FINDINGS: p35 Knockout mice showed no hilar cell death, but moderate mossy fiber sprouting and granule cell dispersion. In the kainate-injected hippocampus, there was an 80% and 85% reduction of GluR2/3- and GluR2/3/calretinin-positive hilar neurons, respectively, and dense mossy fiber sprouting and significant granule cell dispersion. In the contralateral hippocampus there was a 52% loss of GluR2/3-, but only a 20% loss of GluR2/3-calretinin-immunoreactive presumptive mossy cells, and granule cell dispersion; no mossy fiber sprouting was observed. SIGNIFICANCE: These results indicate a probable lack of causality between mossy cell death and mossy fiber sprouting. PMID- 21635233 TI - New concepts in classification of the epilepsies: entering the 21st century. AB - Concepts and terminology for classifying seizures and epilepsies have, until recently, rested on ideas developed nearly a century ago. In order for clinical epilepsy and practice to benefit fully from the major technological and scientific advances of the last several years, advances that are revolutionizing our understanding and treatment of the epilepsies, it is necessary to break with the older vocabulary and approaches to classifying epilepsies and seizures. The Commission on Classification and Terminology made specific recommendations to move this process along and ensure that classification will reflect the best knowledge, will not be arbitrary, and will ultimately serve the purpose of improving clinical practice as well as research on many levels. The recommendations include new terms and concepts for etiology and seizure types as well as abandoning the 1989 classification structure and replacing it instead with a flexible multidimensional approach in which the most relevant features for a specific purpose can be emphasized. This is not a finished product and will take yet more time to achieve. Waiting any longer, however, would be a disservice to patient care and will continue the longstanding frustrations with the earlier system which, at this point in time, can be viewed as both antiquated and arbitrary. PMID- 21635232 TI - Genomic microdeletions associated with epilepsy: not a contraindication to resective surgery. AB - PURPOSE: Several recent reports of genomic microdeletions in epilepsy will generate further research; discovery of more microdeletions and other important classes of variants may follow. Detection of such genetic abnormalities in patients being evaluated for surgical treatment might raise concern that a genetic defect, possibly widely expressed in the brain, will affect surgical outcome. METHODS: A reevaluation was undertaken of clinical presurgical data, histopathology of surgical specimen, and postsurgical outcome in patients with mesial temporal lobe epilepsy (MTLE) who have had surgical treatment for their drug-resistant seizures, and who have been found to have particular genomic microdeletions. KEY FINDINGS: Three thousand eight hundred twelve patients with epilepsy were genotyped and had a genome-wide screen to identify copy number variation. Ten patients with MTLE, who had resective epilepsy surgery, were found to have 16p13.11 microdeletions or other microdeletions >1 Mb. On histopathology, eight had classical hippocampal sclerosis (HS), one had nonspecific findings, and one had a hamartoma. Median postsurgical follow-up time was 48 months (range 10 156 months). All patients with HS were seizure-free after surgery, International League Against Epilepsy (ILAE) outcome class 1, at last follow-up; the patient with nonspecific pathology had recurrence of infrequent seizures after 7 years of seizure freedom. The patient with a hamartoma never became seizure-free. SIGNIFICANCE: Large microdeletions can be found in patients with "typical" MTLE. In this small series, patients with MTLE who meet criteria for resective surgery and harbor large microdeletions, at least those we have detected, can have a good postsurgical outcome. Our findings add to the spectrum of causal heterogeneity of MTLE + HS. PMID- 21635234 TI - First-ever, door-to-door cross-sectional representative study in Prey Veng province (Cambodia). AB - PURPOSE: To estimate the lifetime prevalence of epilepsy in Prey Veng province (Cambodia). METHODS: Door-to-door screening was performed using a random cluster survey whereby all people >1 year of age were screened for epilepsy by using a validated and standardized questionnaire for epilepsy in tropical countries. Suspected epilepsy patients identified by the questionnaire were revisited and examined by epileptologists. The confirmation of epilepsy was based on an in depth clinical examination. Electroencephalograms were recorded at the community dispensary. KEY FINDINGS: Five hundred three potential epilepsy cases were identified from 16,510 screened subjects, and 96 were diagnosed to have epilepsy. An overall prevalence of 5.8 per 1,000 [95% confidence interval (CI) 4.6-7.0 per 1,000] was obtained. Generalized epilepsy (76%) was more common than partial epilepsy (12.5%). Three cases were of generalized myoclonic epilepsy (3.1%) and one case each (1.0%) were of absence and olfactory partial epilepsy. Six cases (5.2%) had more than one seizure type [one case with absence + generalized tonic clonic (GTC), one case each with GTC + partial seizures with secondary generalization and absence + generalized myoclonic seizures and absence + simple partial seizures, and two cases with GTC + complex partial seizures]. Electroencephalography (EEG) studies revealed spike and wave discharges in 43.8%, focal spikes in 21.0%, generalized slow waves in 19.2%, and generalized slowing of background in 15.7%. SIGNIFICANCE: This is the first population-based study in Cambodia that had epilepsy as a primary objective, and compared to Western and neighboring countries it shows a lower prevalence. PMID- 21635235 TI - Memory processes and prefrontal network dysfunction in cryptogenic epilepsy. AB - PURPOSE: Impaired memory performance is the most frequently reported cognitive problem in patients with chronic epilepsy. To examine memory deficits many studies have focused on the role of the mesiotemporal lobe, mostly with hippocampal abnormalities. However, the role of the prefrontal brain remains unresolved. To investigate the neuronal correlates of working memory dysfunction in patients without structural lesions, a combined study of neurocognitive assessment, hippocampal and cerebral volumetry, and functional magnetic resonance imaging of temporal and frontal memory networks was performed. METHODS: Thirty six patients with cryptogenic localization-related epilepsy and 21 healthy controls underwent neuropsychological assessment of intelligence (IQ) and memory. On T(1) -weighted images obtained by 3-Tesla magnetic resonance imaging (MRI), volumetry of the hippocampi and the cerebrum was performed. Functional MRI (fMRI) was performed with a novel picture encoding and Sternberg paradigm that activated different memory-mediating brain regions. Functional connectivity analysis comprised cross-correlation of signal time-series of the most strongly activated regions involved in working memory function. KEY FINDINGS: Patients with epilepsy displayed lower IQ values; impaired transient aspects of information processing, as indicated by lower scores on the digit-symbol substitution test (DSST); and decreased short-term memory performance relative to healthy controls, as measured with the Wechsler Adult Intelligence Scale subtests for working memory, and word and figure recognition. This could not be related to any hippocampal volume changes. No group differences were found regarding volumetry or fMRI-derived functional activation. In the Sternberg paradigm, a network involving the anterior cingulate and the middle and inferior frontal gyrus was activated. A reduced strength of four connections in this prefrontal network was associated with the DSST and word recognition performance in the patient group. SIGNIFICANCE: Deficits in the processes involved in transient working memory, and to a lesser extent in short-term memory, in patients with localization-related epilepsy of both temporal and extratemporal origin cannot be attributed to hippocampal atrophy or function only, but are also related to reduced functional connectivity in the prefrontal brain. Because patients with symptomatic lesions or mesiotemporal sclerosis were excluded from this study, the results cannot be explained by structural lesions. Therefore, the current findings highlight the influence of epilepsy on the prefrontal network integrity as a possible underlying problem of memory impairment. PMID- 21635236 TI - Perampanel: a novel, orally active, noncompetitive AMPA-receptor antagonist that reduces seizure activity in rodent models of epilepsy. AB - PURPOSE: To assess the pharmacology of perampanel and its antiseizure activity in preclinical models. Perampanel [2-(2-oxo-1-phenyl-5-pyridin-2-yl-1,2 dihydropyridin-3-yl) benzonitrile] is a novel, orally active, prospective antiepileptic agent currently in development for refractory partial-onset seizures. METHODS: Perampanel pharmacology was assessed by examining changes in intracellular free Ca(2+) ion concentration ([Ca(2+) ](i) ) in primary rat cortical neurones, and [(3) H]perampanel binding to rat forebrain membranes. Antiseizure activity of orally administered perampanel was examined in amygdala kindled rats and in mice exhibiting audiogenic, maximal electroshock (MES) induced, pentylenetetrazole (PTZ) -induced, or 6 Hz-induced seizures. KEY FINDINGS: In cultured rat cortical neurones, perampanel inhibited alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-induced increases in [Ca(2+) ](i) (IC(50) 93 nm vs. 2 MUm AMPA). Perampanel had a minimal effect on N-methyl-d aspartate (NMDA)-induced increases in [Ca(2+) ](i) , and only at a high concentration (30 MUm). [(3) H]Perampanel binding to rat forebrain membranes was not significantly displaced by glutamate or AMPA but was displaced by the noncompetitive AMPA receptor antagonists CP465022 (K(i) 11.2 +/- 0.8 nm) and GYKI52466 (K(i) 12.4 +/- 1 MUm). In mice, perampanel showed protective effects against audiogenic, MES-induced, and PTZ-induced seizures (ED(50) s 0.47, 1.6, and 0.94 mg/kg, respectively). Perampanel also inhibited 6 Hz electroshock induced seizures when administered alone or in combination with other antiepileptic drugs (AEDs). In amygdala-kindled rats, perampanel significantly increased afterdischarge threshold (p<0.05 vs. vehicle), and significantly reduced motor seizure duration, afterdischarge duration, and seizure severity recorded at 50% higher intensity than afterdischarge threshold current (p<0.05 for all measures vs. vehicle). Perampanel caused dose-dependent motor impairment in both mice (TD(50) 1.8 mg/kg) and rats (TD(50) 9.14 mg/kg), as determined by rotarod tests. In mice, the protective index (TD(50) in rotarod test/ED(50) in seizure test) was 1.1, 3.8, and 1.9 for MES-induced, audiogenic, and PTZ-induced seizures, respectively. In rat, dog, and monkey, perampanel had a half-life of 1.67, 5.34, and 7.55 h and bioavailability of 46.1%, 53.5%, and 74.5%, respectively. SIGNIFICANCE: These data suggest that perampanel is an orally active, noncompetitive, selective AMPA receptor antagonist with potential as a broad spectrum antiepileptic agent. PMID- 21635237 TI - Anomalous levels of Cl- transporters cause a decrease of GABAergic inhibition in human peritumoral epileptic cortex. AB - PURPOSE: Several factors contribute to epileptogenesis in patients with brain tumors, including reduced gamma-aminobutyric acid (GABA)ergic inhibition. In particular, changes in Cl(-) homeostasis in peritumoral microenvironment, together with alterations of metabolism, are key processes leading to epileptogenesis in patients afflicted by glioma. It has been recently proposed that alterations of Cl(-) homeostasis could be involved in tumor cell migration and metastasis formation. In neurons, the regulation of intracellular Cl(-) concentration ([Cl(-) ](i) ) is mediated by NKCC1 and KCC2 transporters: NKCC1 increases while KCC2 decreases [Cl(-) ](i) . Experiments were thus designed to investigate whether, in human epileptic peritumoral cortex, alterations in the balance of NKCC1 and KCC2 activity may decrease the hyperpolarizing effects of GABA, thereby contributing to epileptogenesis in human brain tumors. METHODS: Membranes from peritumoral cortical tissues of epileptic patients afflicted by gliomas (from II to IV WHO grade) and from cortical tissues of nonepileptic patients were injected into Xenopus oocytes leading to the incorporation of functional GABA(A) receptors. The GABA-evoked currents were recorded using standard two-microelectrode voltage-clamp technique. In addition, immunoblot analysis and immunohistochemical staining were carried out on membranes and tissues from the same patients. KEY FINDINGS: We found that in oocytes injected with epileptic peritumoral cerebral cortex, the GABA-evoked currents had a more depolarized reversal potential (E(GABA) ) compared to those from nonepileptic healthy cortex. This difference of E(GABA) was abolished by the NKCC1 blocker bumetanide or unblocking of KCC2 with the Zn(2+) chelator TPEN. Moreover, Western blot analysis revealed an increased expression of NKCC1, and more modestly, of KCC2 transporters in epileptic peritumoral tissues compared to nonepileptic control tissues. In addition, NKCC1 immunoreactivity was strongly increased in peritumoral cortex with respect to nonepileptic cortex, with a prominent expression in neuronal cells. SIGNIFICANCE: We report that the positive shift of E(GABA) in epileptic peritumoral human cortex is due to an altered expression of NKCC1 and KCC2, perturbing Cl(-) homeostasis, which might lead to a consequent reduction in GABAergic inhibition. These findings point to a key role of Cl(-) transporters KCC2 and NKCC1 in tumor-related epilepsy, suggesting a more specific drug therapy and surgical approaches for the epileptic patients afflicted by brain tumors. PMID- 21635238 TI - Effects of early long-term treatment with antiepileptic drugs on development of seizures and depressive-like behavior in a rat genetic absence epilepsy model. AB - PURPOSE: Depression is most commonly associated with epilepsy. Recent reports have suggested a putative relationship between seizure development and onset of depressive behavior, whereas others proposed that two clinical entities might represent different neuropathologic aspects of the same neurologic disorder. The WAG/Rij rat absence epilepsy model has also been proposed as a suitable model to test antidepressant drugs. We previously reported on a long-term study of two antiepileptic drugs (AEDs) to assess their protective role in absence epileptogenesis. Here, we examined the effects of long-term treatment with several AEDs on absence seizure development and onset of depressive-like behavior in WAG/Rij rats at different ages, using a forced swimming test (FST). METHODS: Animals were divided into one untreated control group and four test groups, given ethosuximide, levetiracetam, zonisamide, or carbamazepine. Electroencephalography (EEG) readings were recorded at 6.5 months of age. KEY FINDINGS: Ethosuximide treated animals showed significant reductions in recorded spike-wave discharges (SWDs), and FST immobility time (IT) compared with untreated same age controls. However, zonisamide- and carbamazepine-treated animals had IT values similar to those of controls, but only zonisamide significantly decreased absence seizure development. Carbamazepine increased SWD incidence. Levetiracetam also protected against seizure development, while augmenting IT, suggesting a prodepressive effect. SIGNIFICANCE: Although treatment with ethosuximide, levetiracetam, or zonisamide reduced appearance of SWDs in WAG/Rij rats, this was not generally linked to a reduced onset of depressive characteristics, as assessed by FST. Therefore, expression of depressive-like behavior seems unrelated to seizure control in this model. Some possible alternative explanations for the observed data are discussed. PMID- 21635239 TI - Homeostatic increase in excitability in area CA1 after Schaffer collateral transection in vivo. AB - PURPOSE: Epilepsy is a significant long-term consequence of traumatic brain injury (TBI) and is likely to result from multiple mechanisms. One feature that is common to many forms of TBI is denervation. We asked whether chronic partial denervation in vivo would lead to a homeostatic increase in the excitability of a denervated cell population. METHODS: To answer this question, we took advantage of the unique anatomy of the hippocampus where the input to the CA1 neurons, the Schaffer collaterals, could be transected in vivo with preservation of their outputs and only minor cell death. KEY FINDINGS: We observed a delayed increase in neuronal excitability, as apparent in extracellular recordings from hippocampal brain slices prepared 14 days (but not 3 days) post lesion. Although population spikes in slices from control and lesioned animals were comparable under resting conditions, application of solutions that were mildly proconvulsive (high K(+) , low Mg(2+) , low concentrations of bicuculline) produced increases in the number of population spikes in slices from lesioned rats, but not in slices from unlesioned sham controls. Denervation did not produce changes in several markers of gamma-aminobutyric acid (GABA)ergic synaptic inhibition, including the number of GABAergic neurons, alpha1 GABA(A) receptor subunits, the vesicular GABA transporter, or miniature inhibitory postsynaptic currents. SIGNIFICANCE: We conclude that chronic partial denervation does lead to a delayed homeostatic increase in neuronal excitability, and may, therefore, contribute to the long-term neurologic consequences of TBI. PMID- 21635240 TI - 1H MRSI and social communication deficits in pediatric complex partial seizures. AB - PURPOSE: To investigate relationships between regional brain metabolites, social communication deficits, and seizure frequency in children and adolescents with cryptogenic epilepsy with complex partial seizures (CPS). METHODS: In 12 children and adolescents with CPS and 23 age- and gender-matched healthy controls, we acquired proton magnetic resonance spectroscopic imaging (MRSI) at 1.5 T and 30 ms echo-time from bilateral inferior frontal and superior temporal gyri, regions associated with social communication deficits. Videotaped speech samples of all the subjects were coded for social communication deficits and parents provided information on seizure frequency. KEY FINDINGS: Four MRSI findings emerged in right inferior frontal gyrus. N-acetyl-aspartate (NAA) plus N-acetyl-aspartyl glutamate (NAAG)--together called "tNAA"--was 11.4% lower in patients with CPS than in controls. Choline-compounds (Cho) were 15.4% lower in CPS than in controls. Within CPS, higher tNAA was associated with more frequent seizures and abnormal social communication. SIGNIFICANCE: Localization of findings to right inferior frontal cortex supports the involvement of this area in social communication deficits and may be related to atypical lateralization of expressive language in pediatric epilepsy. Lower levels of tNAA and Cho may indicate local neuronal or glial damage or underpopulation due to excitotoxicity or other causes. The sensitivity of tNAA to seizure frequency suggests effects of ongoing CPS on neuronal and glial function in this brain region. PMID- 21635241 TI - Upregulation of adenosine kinase in astrocytes in experimental and human temporal lobe epilepsy. AB - PURPOSE: Adenosine kinase (ADK) represents the key metabolic enzyme for the regulation of extracellular adenosine levels in the brain. In adult brain, ADK is primarily present in astrocytes. Several lines of experimental evidence support a critical role of ADK in different types of brain injury associated with astrogliosis, which is also a prominent morphologic feature of temporal lobe epilepsy (TLE). We hypothesized that dysregulation of ADK is an ubiquitous pathologic hallmark of TLE. METHODS: Using immunocytochemistry and Western blot analysis, we investigated ADK protein expression in a rat model of TLE during epileptogenesis and the chronic epileptic phase and compared those findings with tissue resected from TLE patients with mesial temporal sclerosis (MTS). KEY FINDINGS: In rat control hippocampus and cortex, a low baseline expression of ADK was found with mainly nuclear localization. One week after the electrical induction of status epilepticus (SE), prominent up-regulation of ADK became evident in astrocytes with a characteristic cytoplasmic localization. This increase in ADK persisted at least for 3-4 months after SE in rats developing a progressive form of epilepsy. In line with the findings from the rat model, expression of astrocytic ADK was also found to be increased in the hippocampus and temporal cortex of patients with TLE. In addition, in vitro experiments in human astrocyte cultures showed that ADK expression was increased by several proinflammatory molecules (interleukin-1beta and lipopolysaccharide). SIGNIFICANCE: These results suggest that dysregulation of ADK in astrocytes is a common pathologic hallmark of TLE. Moreover, in vitro data suggest the existence of an additional layer of modulatory crosstalk between the astrocyte-based adenosine cycle and inflammation. Whether this interaction also can play a role in vivo needs to be further investigated. PMID- 21635242 TI - Microstructural and volumetric abnormalities of the putamen in juvenile myoclonic epilepsy. AB - PURPOSE: Patients with juvenile myoclonic epilepsy (JME) show evidence of microstructural white matter (WM) damage of thalamocortical fiber tracts and changes of blood oxygen level dependent (BOLD) signal in a striatothalamocortical network. The objective of the present study was to investigate microstructural and volumetric alterations of the putamen in patients with JME using diffusion tensor imaging (DTI) and conventional magnetic resonance imaging (MRI). METHODS: We performed DTI and MRI for 10 patients with JME and 59 age-matched neurologically healthy volunteers. Evaluation of microstructural damage was investigated using calculation of mean fractional anisotropy (FA) values in a priori regions of interest (ROIs) for the putamen, frontal lobe, and a thalamocortical region, after application of an improved eddy current correction method and a new statistical parametric mapping (SPM)-compatible toolbox incorporating intensive multicontrast FA image registration. Stereologic analysis on MRI was performed to estimate macroscopic volume of the putamen in both cerebral hemispheres for all subjects. KEY FINDINGS: Relative to controls, patients had significantly reduced FA in the frontal lobe (p = 0.01) and thalamocortical fiber WM (p < 0.001). In contrast, putamen FA was bilaterally increased (p = 0.01) and correlated with decreasing putamen volume (r(2) = -0.63, p = 0.004) in patients only. Putamen FA correlated negatively with onset of JME (total: r(2) = -0.50, p = 0.01), duration of JME (r(2) = 0.52, p = 0.01), and thalamocortical fiber FA (r(2) = -0.47, p = 0.01). SIGNIFICANCE: This is the first evidence of combined microstructural and macrostructural putamen abnormalities in patients with JME, with early age of onset and a longer duration of epilepsy being significant predictors for greater architectural alterations. These findings are consistent with studies indicating neurophysiologic abnormalities of frontostriatal networks in patients with JME, and may contribute to explain the frequent presentation of executive dysfunction in these patients. Confirmation and further exploration of the increase in putamen FA in patients with JME is required in larger samples. PMID- 21635243 TI - Effects of lamotrigine and phenytoin on the pharmacokinetics of atorvastatin in healthy volunteers. AB - PURPOSE: Statins and antiepileptic drugs (AEDs) are frequently coprescribed to individuals with hypercholesterolemia and new-onset seizures. Statins are metabolized by the cytochrome P450 (CYP) enzyme system. Interactions between statins and agents that undergo CYP metabolism are common. In this study, the effects of two commonly prescribed AEDs, lamotrigine and phenytoin, with different routes of metabolism (CYP3A4 versus glucuronic acid conjugation) on atorvastatin pharmacokinetics were evaluated. METHODS: Healthy volunteers (n=119) received atorvastatin 40 mg/day for 7 days followed by addition of lamotrigine (target 300 mg/day) or phenytoin (target ~4 mg/kg per day) in this open-label, single-sequence, two-cohort study. Serial pharmacokinetic sampling of atorvastatin was conducted on day 7 of atorvastatin dosing and day 70 of lamotrigine + atorvastatin dosing or day 28 of phenytoin + atorvastatin dosing. Main outcome measures were steady-state area under the curve over the 24-h dosing interval (AUC((0-tau)) ) and maximum concentration (C(max) ) of atorvastatin and its metabolites, 2OH-atorvastatin and 4OH-atorvastatin, in the presence of lamotrigine or phenytoin. KEY FINDINGS: When atorvastatin was administered with lamotrigine compared with when atorvastatin was administered alone, atorvastatin AUC((0-tau)) was within bounds indicating no interaction, whereas C(max) was slightly higher(14%); AUC((0-tau)) and C(max) were 3% and 20% higher, respectively, for 2OH-atorvastatin and 25% and 21% higher, respectively, for 4OH atorvastatin.When atorvastatin was administered with phenytoin compared with when atorvastatin was administered alone, reductions in AUC((0-tau)) and C(max) were observed for atorvastatin (54% and 24%, respectively), 2OH-atorvastatin (53% and 22%, respectively), and 4OH-atorvastatin (44% and 52%, respectively). SIGNIFICANCE: Pharmacokinetics of atorvastatin were not significantly affected by coadministration with lamotrigine. Phenytoin significantly reduced atorvastatin bioavailability. Consistent with the published literature, these data are consonant with the possibility that atorvastatin does not require dose adjustment when coadministered with lamotrigine at doses to 300 mg/day, whereas atorvastatin coadministered with phenytoin may require atorvastatin dose adjustment to maintain atorvastatin exposure. PMID- 21635244 TI - Symptoms of anxiety and depression in childhood absence epilepsy. AB - Childhood absence epilepsy (CAE) has been recently linked to a number of cognitive, behavioral, and emotional disorders. Identification of affective disorders (anxiety and depression) presents unique challenges in pediatric populations, and successful early intervention may significantly improve long term developmental outcomes. The current study examined the specific anxiety and depression symptoms children with CAE experience, and explored the role of disease factors in the severity of their presentation. Forty-five subjects with CAE and 41 healthy matched controls, ages 6-16 years, participated in the study. The Behavior Assessment System for Children (BASC) was completed by parents, and the Anxiety and Depression subscales were used to characterize problems. Item analysis within the subscales revealed that children with CAE demonstrated higher rates of symptoms of anxiety (nervousness and thought rumination) and depression (sadness and crying), as well as more general psychosocial problems including isolation and low self-esteem. Disease duration, intractability, and medication effects were not associated with higher rates of affective problems in this limited patient sample. Screening of patients with CAE for comorbid psychiatric disorders early by focusing on specific symptom profiles unique to this population may enhance overall treatment and developmental outcomes. PMID- 21635245 TI - Hybrid-optimization algorithm for the management of a conjunctive-use project and well field design. AB - Hi-Desert Water District (HDWD), the primary water-management agency in the Warren Groundwater Basin, California, plans to construct a waste water treatment plant to reduce future septic-tank effluent from reaching the groundwater system. The treated waste water will be reclaimed by recharging the groundwater basin via recharge ponds as part of a larger conjunctive-use strategy. HDWD wishes to identify the least-cost conjunctive-use strategies for managing imported surface water, reclaimed water, and local groundwater. As formulated, the mixed-integer nonlinear programming (MINLP) groundwater-management problem seeks to minimize water-delivery costs subject to constraints including potential locations of the new pumping wells, California State regulations, groundwater-level constraints, water-supply demand, available imported water, and pump/recharge capacities. In this study, a hybrid-optimization algorithm, which couples a genetic algorithm and successive-linear programming, is developed to solve the MINLP problem. The algorithm was tested by comparing results to the enumerative solution for a simplified version of the HDWD groundwater-management problem. The results indicate that the hybrid-optimization algorithm can identify the global optimum. The hybrid-optimization algorithm is then applied to solve a complex groundwater management problem. Sensitivity analyses were also performed to assess the impact of varying the new recharge pond orientation, varying the mixing ratio of reclaimed water and pumped water, and varying the amount of imported water available. The developed conjunctive management model can provide HDWD water managers with information that will improve their ability to manage their surface water, reclaimed water, and groundwater resources. PMID- 21635246 TI - Approaches to the simulation of unconfined flow and perched groundwater flow in MODFLOW. AB - Various approaches have been proposed to manage the nonlinearities associated with the unconfined flow equation and to simulate perched groundwater conditions using the MODFLOW family of codes. The approaches comprise a variety of numerical techniques to prevent dry cells from becoming inactive and to achieve a stable solution focused on formulations of the unconfined, partially-saturated, groundwater flow equation. Keeping dry cells active avoids a discontinuous head solution which in turn improves the effectiveness of parameter estimation software that relies on continuous derivatives. Most approaches implement an upstream weighting of intercell conductance and Newton-Raphson linearization to obtain robust convergence. In this study, several published approaches were implemented in a stepwise manner into MODFLOW for comparative analysis. First, a comparative analysis of the methods is presented using synthetic examples that create convergence issues or difficulty in handling perched conditions with the more common dry-cell simulation capabilities of MODFLOW. Next, a field-scale three-dimensional simulation is presented to examine the stability and performance of the discussed approaches in larger, practical, simulation settings. PMID- 21635247 TI - V. Shestakov and the development of modern hydrogeology in Russia. PMID- 21635248 TI - T regulatory cells participate in the control of germinal centre reactions. AB - Germinal centre (GC) reactions are central features of T-cell-driven B-cell responses, and the site where antibody-producing cells and memory B cells are generated. Within GCs, a range of complex cellular and molecular events occur which are critical for the generation of high affinity antibodies. These processes require exquisite regulation not only to ensure the production of desired antibodies, but to minimize unwanted autoreactive or low affinity antibodies. To assess whether T regulatory (Treg) cells participate in the control of GC responses, immunized mice were treated with an anti-glucocorticoid induced tumour necrosis factor receptor-related protein (GITR) monoclonal antibody (mAb) to disrupt Treg-cell activity. In anti-GITR-treated mice, the GC B cell pool was significantly larger compared with control-treated animals, with switched GC B cells composing an abnormally high proportion of the response. Dysregulated GCs were also observed regardless of strain, T helper type 1 or 2 polarizing antigens, and were also seen after anti-CD25 mAb treatment. Within the spleens of immunized mice, CXCR5(+) and CCR7(-) Treg cells were documented by flow cytometry and Foxp3(+) cells were found within GCs using immunohistology. Final studies demonstrated administration of either anti-transforming growth factor-beta or anti-interleukin-10 receptor blocking mAb to likewise result in dysregulated GCs, suggesting that generation of inducible Treg cells is important in controlling the GC response. Taken together, these findings indicate that Treg cells contribute to the overall size and quality of the humoral response by controlling homeostasis within GCs. PMID- 21635249 TI - Expression of mesenchyme-specific gene signatures by follicular dendritic cells: insights from the meta-analysis of microarray data from multiple mouse cell populations. AB - Follicular dendritic cells (FDC) are an important subset of stromal cells within the germinal centres of lymphoid tissues. They are specialized to trap and retain antigen-containing immune complexes on their surfaces to promote B-cell maturation and immunoglobulin isotype class-switching. However, little is known of the cell types from which FDC originate. To address fundamental questions associated with the relationships between FDC and other cell populations, we took advantage of the growing body of publicly available data for transcriptome analysis. We obtained a large number of gene expression data files from a range of different primary mouse cells and cell lines and subjected these data to network-based cluster analysis using BiolayoutExpress(3D) . Genes with related function clustered together in distinct regions of the graph and enabled the identification of transcriptional networks that underpin the functional activity of distinct cell populations. Several gene clusters were identified that were selectively expressed by cells of mesenchymal lineage and contained classic mesenchymal cell markers and extracellular matrix genes including various collagens, Acta2, Bgn, Fbn1 and Twist1. Our analysis showed that FDC also express highly many of these mesenchyme-associated genes. Promoter analysis of the genes comprising the mesenchymal clusters identified several regulatory motifs that are binding sites for candidate transcription factors previously known to be candidate regulators of mesenchyme-specific genes. Together, these data suggest FDC are a specialized mesenchymal cell population within the germinal centres of lymphoid tissues. PMID- 21635250 TI - Effects of growth on geometry of gastrocnemius muscle in children: a three dimensional ultrasound analysis. AB - During development, muscle growth is usually finely adapted to meet functional demands in daily activities. However, how muscle geometry changes in typically developing children and how these changes are related to functional and mechanical properties is largely unknown. In rodents, longitudinal growth of the pennate m. gastrocnemius medialis (GM) has been shown to occur mainly by an increase in physiological cross-sectional area and less by an increase in fibre length. Therefore, we aimed to: (i) determine how geometry of GM changes in healthy children between the ages of 5 and 12 years, (ii) test whether GM geometry in these children is affected by gender, (iii) compare normalized growth of GM geometry in children with that in rats at similar normalized ages, and (iv) investigate how GM geometry in children relates to range of motion of angular foot movement at a given moment. Thirty children (16 females, 14 males) participated in the study. Moment-angle data were collected over a range of angles by rotating the foot from plantar flexion to dorsal flexion at standardized moments. GM geometry in the mid-longitudinal plane was measured using three-dimensional ultrasound imaging. This geometry was compared with that of GM geometry in rats. During growth from 5 to 12 years of age, the mean neutral footplate angle (0 Nm) occurred at -5 degrees (SD 7 degrees ) and was not a function of age. Measured at standardized moments (4 Nm), footplate angles towards plantar flexion and dorsal flexion decreased by 25 and 40%, respectively. In both rats and children, GM muscle length increased proportionally with tibia length. In children, the length component of the physiological cross-sectional area and fascicle length increased by 7 and 5% per year, respectively. Fascicle angle did not change over the age range measured. In children, the Achilles tendon length increased by 6% per year. GM geometry was not affected by gender. We conclude that, whereas the length of GM in rat develops mainly by an increase in physiological cross-sectional area of the muscle, GM in children develops by uniform scaling of the muscle. This effect is probably related to the smaller fascicle angle in human GM, which entails a smaller contribution of radial muscle growth to increased GM muscle length. The net effect of uniform scaling of GM muscle belly causes it to be stiffer, explaining the decrease in range of motion of angular foot movement at 4 Nm towards dorsal flexion during growth. PMID- 21635251 TI - On observation distributions for state space models of population survey data. AB - 1. State space models are starting to replace more simple time series models in analyses of temporal dynamics of populations that are not perfectly censused. By simultaneously modelling both the dynamics and the observations, consistent estimates of population dynamical parameters may be obtained. For many data sets, the distribution of observation errors is unknown and error models typically chosen in an ad-hoc manner. 2. To investigate the influence of the choice of observation error on inferences, we analyse the dynamics of a replicated time series of red kangaroo surveys using a state space model with linear state dynamics. Surveys were performed through aerial counts and Poisson, overdispersed Poisson, normal and log-normal distributions may all be adequate for modelling observation errors for the data. We fit each of these to the data and compare them using AIC. 3. The state space models were fitted with maximum likelihood methods using a recent importance sampling technique that relies on the Kalman filter. The method relaxes the assumption of Gaussian observation errors required by the basic Kalman filter. Matlab code for fitting linear state space models with Poisson observations is provided. 4. The ability of AIC to identify the correct observation model was investigated in a small simulation study. For the parameter values used in the study, without replicated observations, the correct observation distribution could sometimes be identified but model selection was prone to misclassification. On the other hand, when observations were replicated, the correct distribution could typically be identified. 5. Our results illustrate that inferences may differ markedly depending on the observation distributions used, suggesting that choosing an adequate observation model can be critical. Model selection and simulations show that for the models and parameter values in this study, a suitable observation model can typically be identified if observations are replicated. Model selection and replication of observations, therefore, provide a potential solution when the observation distribution is unknown. PMID- 21635252 TI - Interleukin-17 and interleukin-18 levels in saliva and plasma of patients with chronic periodontitis. AB - BACKGROUND AND OBJECTIVE: This study was planned to investigate whether patients with chronic periodontitis exhibit different salivary and/or plasma concentrations of interleukin (IL)-17 and IL-18 compared with clinically healthy subjects. MATERIAL AND METHODS: Whole saliva and blood samples, together with full-mouth clinical periodontal recordings, were obtained from 22 otherwise healthy untreated nonsmokers with chronic periodontitis and from 21 systemically and periodontally healthy control subjects. The concentrations of IL-17 and IL-18 in saliva and plasma were determined using ELISAs. RESULTS: The healthy control group exhibited significantly lower values in all clinical periodontal measurements (p < 0.001). The salivary concentration of IL-17 was significantly lower, and that of IL-18 significantly higher, in patients from the chronic periodontitis group compared with healthy control subjects (p = 0.025 and p = 0.009, respectively). Plasma IL-17 and IL-18 concentrations were similar in the two study groups (p > 0.05). CONCLUSION: Within the limits of the present study, it may be suggested that an elevated salivary IL-18 level in untreated nonsmoker chronic periodontitis patients has the potential to be a biomarker for periodontal tissue destruction. PMID- 21635253 TI - Current drug discovery strategies for treatment of hepatitis C virus infection. AB - OBJECTIVES: Hepatitis C virus (HCV) infection represents a major worldwide-health problem. The current standard of care is combination therapy with pegylated interferon and ribavirin, which achieves a successful response in only approximately 40% of genotype I patients. KEY FINDINGS: The biology of HCV infection has been under intensive research and important progress has been made in understanding the replication cycle of the virus. Several therapeutic targets have been under investigation, such as NS3 protease, NS4A replicase and NS5B polymerase. New potential targets, such as NS2 protease, as well as CD-81 and claudin-1 entry co-receptors, have also been identified. SUMMARY: Clinical evaluations of drug candidates targeting NS3 protease, NS4A cofactor, and NS5B polymerase have demonstrated the potential of developing small molecules that interfere with the replication of the virus. Additional issues, including genotype coverage, resistant mutations, and combination therapy represent major challenges for future drug discovery efforts. PMID- 21635254 TI - Design of clinically useful macromolecular iron chelators. AB - OBJECTIVES: In recent years, macromolecular iron chelators have received increasing attention as human therapeutic agents. The objectives of this article are: one, to discuss the factors which should be considered when designing iron binding macromolecules as human therapeutic agents, and two, to report recent achievements in the design and synthesis of appropriate macromolecular chelators that have resulted in the production of a number of agents with therapeutic potential. KEY FINDINGS: Macromolecular drugs exhibit unique pharmaceutical properties that are fundamentally different from their traditional small-molecule counterparts. By virtue of their high-molecular-weight characteristics, many are confined to extracellular compartments, for instance, the serum and the gastrointestinal tract. In addition, they have potential for topical administration. Consequently, these macromolecular drugs are free from many of the toxic effects that are associated with their low-molecular-weight analogues. SUMMARY: The design and synthesis of macromolecular iron chelators provides a novel aspect to chelation therapy. 3-Hydroxypyridin-4-one hexadentate-based macromolecular chelators have considerable potential for the development of new treatments for iron overload and for topical treatment of infection. PMID- 21635255 TI - Multivesicular liposome (MVL) sustained delivery of a novel synthetic cationic GnRH antagonist for prostate cancer treatment. AB - OBJECTIVES: Multivesicular liposomes (MVLs) are often used as an appropriate carrier for delivering peptides due to high drug loading, relative stability and extended-release behaviour. However, when cationic amphipathic peptides are involved, some challenges may be encountered, including instability of multiple emulsions due to interaction between peptides and lipid membranes (electrostatic and hydrophobic interaction). LXT-101, a cationic amphipathic peptide, is a novel antagonist of gonadotropin-releasing hormone (GnRH) for prostate cancer treatment. The purpose of the current research was to explore simple methods of determining the interaction between peptide and lipid bilayer and to prepare MVLs of LXT-101 (DepoLXT-101) by the modified DepoFoam technique. METHODS: The anionic surfactants were added in the process of DepoLXT-101 preparation in order to minimize the effect of instability resulting from cationic peptides. KEY FINDINGS: DepoLXT-101 was obtained with good efficiency and reproduction. The integrity of encapsulated peptide was maintained as shown by RP-HPLC. DepoLXT-101 particles were characterized by morphology and particle size distribution and in vitro release was also investigated. The release behaviour in vitro in medium of sodium chloride at 37 degrees C showed that 70-90% of LXT-101 was released slowly from MVLparticles over 11 days. According to the fitting results of Ritgar-Pepps model, the in-vitro release of DepoLXT-101 was mainly governed by Fick's diffusion. CONCLUSIONS: The data obtained from in-vivo study indicated that a sustained anticancer effect can be achieved over a 7-day period with subcutaneous administration of DepoLXT-101 in rats. PMID- 21635256 TI - Quantitative analysis of pseudopolymorphic transformation of imidafenacin by application of a novel combination of near-infrared spectroscopy and a humidity controlled 96-well plate. AB - OBJECTIVES: This study was conducted to test whether our novel combination (i.e. microanalysis) of near-infrared (NIR) spectroscopy and humidity-controlled 96 well plate, reported in a previous issue of this journal, can be successfully applied to quantitative evaluation of the pseudopolymorphic transformation of imidafenacin. METHODS: Sample powders of a drug compound were placed in a humidity-controlled 96-well plate containing various saturated salt solutions, and stored at 35 degrees C. NIR spectra were collected using a Fourier transform NIR spectrometer in combination with a fiber-optic probe. The actual hydrate contents of samples were determined by using thermal gravimetric analysis (TGA). A series of sets of NIR spectra and TGA data were used to establish a calibration model with which to predict the contents of monohydrate by partial least-squares regression (PLS). KEY FINDINGS: The PLS calibration model analysis showed that the plots of NIR predicted values to the actual values gave a straight line with correlation coefficients of 0.9868 and 0.9936, respectively, for the 96-well plate and glass-bottle uses. The model using the 96-well plate was therefore able to predict the transformation of imidafenacin in one form to another form as quantitatively as the conventional model using glass bottles. CONCLUSION: The present study confirmed that our microanalysis quantitatively predicts the pseudopolymorphic transformation of our tested drug in small amounts and suggests that this particular method is a simple and convenient one that may be efficiently applied to polymorphic and pseudopolymorphic studies of a hygroscopic drug candidate, at even earlier development stages than the conventional method using glass bottles. PMID- 21635257 TI - Metronidazole leads to enhanced uptake of imatinib in brain, liver and kidney without affecting its plasma pharmacokinetics in mice. AB - OBJECTIVES: The pharmacokinetic interaction between metronidazole, an antibiotic antiparasitic drug used to treat anaerobic bacterial and protozoal infections, and imatinib, a CYP3A4, P-glycoprotein substrate kinase inhibitor anticancer drug, was evaluated. METHODS: Male imprinting control region mice were given 50 mg/kg imatinib PO (control group) or 50 mg/kg imatinib PO, 15 min after 40 mg/kg PO metronidazole (study group). Imatinib plasma, brain, kidney and liver concentrations were measured by HPLC and non-compartmental pharmacokinetic parameters estimated. KEY FINDINGS: Metronidazole coadministration resulted in a double-peak imatinib disposition profile. The maximum concentration (C(max)) decreased by 38%, the area under the curve (AUC(0-infinity)) decreased by 14% and the time to C(max) (T(max)) was earlier (50%) in plasma. Apparent volume of distribution (V(SS)/F) and oral clearance (Cl/F) increased by 21% and 17%, respectively. Imatinib tissue penetration was higher after metronidazole coadministration, with 1.7 and 2.1-fold AUC(0-infinity) increases in liver and kidney, respectively. Metronidazole increased imatinib's tissue-to-plasma AUC(0 infinity) ratio in liver from 2.29 to 4.53 and in kidney from 3.04 to 7.57, suggesting higher uptake efficiency. Brain C(max) was 3.9-fold higher than control and AUC(0-t last) was 2.3-fold greater than plasma (3.5% in control group). No tissue-plasma concentration correlation was found. CONCLUSIONS: Metronidazole slightly decreased imatinib systemic exposure but enhanced liver, kidney and brain penetration, probably due to metronidazole-mediated inhibition of P-glycoprotein and other efflux transporters. The high brain exposure opens possibilities for treatment of glioma and glioblastoma. Renal and hepatic functions may need to be monitored due to potential renal and hepatic toxicity. PMID- 21635258 TI - Gnaphaliin A and B relax smooth muscle of guinea-pig trachea and rat aorta via phosphodiesterase inhibition. AB - OBJECTIVES: To explore the relaxant mechanism of action of gnaphaliin A and gnaphaliin B in guinea-pig trachea and rat aorta, and to investigate the theoretical and experimental phosphodiesterase (PDE) inhibitory activity of these flavones. METHODS: The relaxant effect and the inhibition of calcium chloride induced contractions of both flavones were evaluated on guinea-pig trachea and rat aorta rings. The PDE inhibitory activity was evaluated using a cyclic nucleotide PDE colorimetric assay kit with cAMP and cGMP as substrates. The docking analysis was carried out with AutoDock4 software and X-ray structure of PDE type 5. The activity of both gnaphaliins was compared with the activity of sildenafil, rolipram, aminophylline, IBMX and enoximone. KEY FINDINGS: Gnaphaliin A and B were more actives as relaxants on rat aorta than guinea-pig trachea. They were less potent in the relaxation of guinea-pig trachea and rat aorta than sildenafil, but they were equal or more potent than the other PDE inhibitors tested. The relaxant effect of these flavones was potentiated by nitroprusside and forskolin, and blocked by 1H-[1,2,4]-oxadiazolo[4,3-a]quinoxalin-1-one but not by 2',5'-dideoxyadenosine in guinea-pig trachea. L-NAME did not modify the relaxant effect of gnaphaliins. Gnaphaliins were more potent as PDE inhibitors when cGMP was used as substrate. Docking analysis revealed that gnaphaiins bind to the same binding site of sildenafil at PDE type 5. CONCLUSIONS: The results suggest that the main relaxant mechanism of action of gnaphaliin A and B is inhibition of PDEs with a preference to inhibit the degradation of cGMP. The docking study suggested that these flavones bind with high specificity to the same binding site of sildenafil at PDE type 5. PMID- 21635259 TI - Structure activity relationship, acute toxicity and cytotoxicity of antimycobacterial neolignan analogues. AB - OBJECTIVES: The study's aims were to evaluate the antimycobacterial activity of 13 synthetic neolignan analogues and to perform structure activity relationship analysis (SAR). The cytotoxicity of the compound 2-phenoxy-1-phenylethanone (LS 2, 1) in mammalian cells, such as the acute toxicity in mice, was also evaluated. METHODS: The extra and intracellular antimycobacterial activity was evaluated on Mycobacterium tuberculosis H37Rv. Cytotoxicity studies were performed using V79 cells, J774 macrophages and rat hepatocytes. Additionally, the in-vivo acute toxicity was tested in mice. The SAR analysis was performed by Principal Component Analysis (PCA). KEY FINDINGS: Among the 13 analogues tested, LS-2 (1) was the most effective, showing promising antimycobacterial activity and very low cytotoxicity in V79 cells and in J774 macrophages, while no toxicity was observed in rat hepatocytes. The selectivity index (SI) of LS-2 (1) was 91 and the calculated LD50 was 1870 mg/kg, highlighting the very low toxicity in mice. SAR analysis showed that the highest electrophilicity and the lowest molar volume are physical-chemical characteristics important for the antimycobacterial activity of the LS-2 (1). CONCLUSIONS: LS-2 (1) showed promising antimycobacterial activity and very weak cytotoxicity in cell culture, as well as an absence of toxicity in primary culture of hepatocytes. In the acute toxicity study there was an indication of absence of toxicity on murine models, in vivo. PMID- 21635260 TI - Sildenafil improves diabetic vascular activity through suppressing endothelin receptor A, iNOS and NADPH oxidase which is comparable with the endothelin receptor antagonist CPU0213 in STZ-injected rats. AB - OBJECTIVES: Abnormal vascular activity in diabetes is related not only to impaired nitric oxide bioavailability but also to inflammatory cytokines, endothelin A receptor (ET(A) ) activation and NADPH oxidase in the vasculature. The potential role of sildenafil in improving vascular function was investigated. Its action was likely blocking upregulated ET(A) and NADPH oxidase, and was compared with the endothelin receptor antagonist CPU0213. METHODS: Diabetes was induced by single-dose administration of streptozotocin (65 mg/kg, i.p.) to rats and the vascular activity of the thoracic aorta was measured. KEY FINDINGS: An increase in contractile tone to phenylephrine and a decrease in relaxant tone to acetylcholine was found in the thoracic aorta. Oxidative stress was evident by increased malondialdehyde and reduced glutathione peroxidase levels in serum and upregulation of ET(A), MMP-9 (matrix metalloproteinase-9), inducible nitric oxide synthase and NADPH oxidase p67(phox) were found in the vascular wall. The vascular abnormalities and abnormal biomarkers were attenuated significantly by either sildenafil or CPU0213 along with an improvement of nitric oxide bioavailability and vascular activity. CONCLUSIONS: Improvement of diabetic vascular abnormal activity by sildenafil results from its suppression of activation of ET(A) and NADPH oxidase in the vasculature, and these actions are comparable with those of the endothelin receptor antagonist CPU0213. PMID- 21635261 TI - Hepato-/reno-protective activity of Chinese prescription Kangen-karyu through inhibition of AGE formation and fibrosis-related protein expression in type 2 diabetes. AB - OBJECTIVES: This study was conducted to examine whether Kangen-karyu, a Chinese prescription, has an ameliorative effect on diabetes-induced alterations such as advanced glycation endproduct (AGE) formation or the fibrotic response in liver and kidney of type 2 diabetic db/db mice. METHODS: Kangen-karyu (100 or 200 mg/kg body weight/day, p.o.) was administered every day for 18 weeks to db/db mice, and its effect was compared with vehicle-treated db/db and m/m mice. KEY FINDINGS: The administration of Kangen-karyu decreased the elevated serum glucose concentration in db/db mice. The increased serum creatinine and urea nitrogen levels, which reflect renal dysfunction in db/db mice, were significantly lowered by Kangen-karyu administration. The db/db mice exhibited the up-regulation of AGEs and its receptor expression in liver and kidney; however, Kangen-karyu treatment significantly reduced expression except for the receptor. Moreover, the augmented expressions of fibrosis-related proteins, transforming growth factor (TGF)-beta1, fibronectin and collagen IV were down-regulated by Kangen-karyu administration. CONCLUSIONS: These results provide important evidence that Kangen karyu exhibits a pleiotropic effect on AGE formation and fibrosis-related parameters, representing hepatoprotective and renoprotective effects against the development of diabetic complications in type 2 diabetic db/db mice. PMID- 21635262 TI - Lapachol suppresses cell proliferation and secretion of interleukin-6 and plasminogen activator inhibitor-1 of fibroblasts derived from hypertrophic scars. AB - OBJECTIVES: The pathogenesis and therapy of hypertrophic scar have not yet been established. Our aim was to investigate the antiproliferative and antisecretory effects of lapachol, isolated from the stem bark of Avicennia rumphiana Hall. f., on hypertrophic scar fibroblasts. METHODS: The effects of lapachol on hypertrophic scar fibroblast proliferation were measured using the MTT assay, cell-cycle analyses and lactate dehydrogenase assays. The type I collagen alpha chain (COL1A1), interleukin-6 (IL-6) and plasminogen activator inhibitor-1 (PAI 1) mRNA and/or protein levels of hypertrophic scar-fibroblasts were quantitated by real-time PCR and ELISA. KEY FINDINGS: Lapachol at 25 and 50 um significantly inhibited the in vitro proliferation of hypertrophic scar fibroblasts, but not fibroblasts from non-lesional skin sites. In addition, lapachol had no apparent effect on cell cycle and lactate dehydrogenase activity in conditioned medium from lapachol-treated hypertrophic scar fibroblasts was nearly equal to that in medium from vehicle-treated cells. Lapachol treatment also inhibited COL1A1 and PAI-1 mRNA levels in hypertrophic scar fibroblasts, but did not affect IL-6 mRNA levels. The protein levels of IL-6 and PAI-1 in conditioned medium from hypertrophic scar fibroblasts treated with 50 um lapachol were lower than those from vehicle-treated hypertrophic scar fibroblasts. CONCLUSIONS: Lapachol decreased the proliferation rate of hypertrophic scar fibroblasts. As IL-6 and PAI-1 secretion was also lowered in lapachol-treated hypertrophic scar fibroblasts, our findings suggested that lapachol may have suppressed extracellular matrix hyperplasia in wound healing and possibly alleviated the formation of hypertrophic scar. PMID- 21635263 TI - Sulfation of selected mono-hydroxyflavones by sulfotransferases in vitro: a species and gender comparison. AB - OBJECTIVES: Sulfation via sulfotransferases is an important metabolic pathway contributing to the low bioavailability of flavonoids. This study aims to characterize the sulfation of mono-hydroxyflavones (MHFs) to obtain useful information on structure-metabolizing relationships in animal species and gender differences. METHODS: Three representative MHFs, namely, 7-, 6- and 4'-MHF, were studied by incubating each MHF at different concentrations with various liver S9 fractions (mouse, rat, dog and human). KEY FINDINGS: One mono-sulfate was identified for each MHF. 7-MHF and 4'-MHF usually have greater sulfations than 6 MHF. Regardless of whether the S9 fraction came from a male or female, there was a difference in sulfation in the species observed for all MHFs; the highest activity of sulfotransferases was in dog S9. Furthermore, gender differences affect sulfation of MHFs significantly. In rats, all sulfations for the three MHFs were higher in males than that in females while the opposite was observed in mice. CONCLUSIONS: Regiospecific, species and gender dependence exist in the sulfonation of all selected MHFs. PMID- 21635264 TI - Topical anti-inflammatory effect of hypocholesterolaemic drugs. AB - OBJECTIVES: The topical anti-inflammatory effect of simvastatin, atorvastatin, pravastatin, ezetimibe and combined ezetimibe + simvastatin was investigated, using the croton oil model of ear oedema in mice. METHODS: Simvastatin, atorvastatin, pravastatin, ezetimibe and ezetimibe + simvastatin combination (dissolved in 20 ul of 70% acetone) were topically applied simultaneously with croton oil (200 ug/ear, dissolved in 20 ul of 70% acetone) at the inner surface of each ear. Ear oedema and myeloperoxidase activity, indicative of polymorphonuclear cell migration, were assessed 6 h after inflammatory stimuli. KEY FINDINGS: It was found that statins can act as topical anti-inflammatories, but the pharmacological effect is dependent on statin polarity. At 0.3 mg/ear inhibition of ear oedema was 79%, 67% and 40% for simvastatin, atorvastatin and pravastatin, respectively. Simvastatin and atorvastatin also remarkably diminished myeloperoxidase activity, even at low concentrations (0.03 mg/ear). Pravastatin, the most polar statin, however, did not cause any reduction in ear oedema or myeloperoxidase activity at low doses. The order of topical anti inflammatory activity was pravastatin < < < atorvastatin <= simvastatin. Ezetimibe, another hypocholesterolaemic drug, also presented anti-inflammatory effects, inhibiting ear oedema by 64% at 0.3 mg/ear. However, when used in combination with simvastatin, no further beneficial effect was observed. CONCLUSIONS: These results consistently support current evidence showing that statins can be used for treatment of dermatological disorders. Polarity of the molecule, however, is a factor that should be considered before recommending use. PMID- 21635265 TI - No effect of co-administered antiepileptic drugs on in-vivo protein binding parameters of valproic acid in patients with epilepsy. AB - OBJECTIVES: The aim of this study was to establish the population protein binding parameters of valproic acid (VPA) in patients with epilepsy receiving VPA monotherapy and those receiving VPA combined with other antiepileptic drugs. METHODS: One hundred and thirty nine data sets from 63 Japanese patients with epilepsy were analysed. These patients were separated into two groups: VPA monotherapy and VPA combined with other binding-sensitive antiepileptic drugs, including phenytoin, clonazepam, clobazam, carbamazepine and phenobarbital (VPA polytherapy). The population protein-binding parameters of VPA were obtained by non-linear least-squares method in each group. KEY FINDINGS: The mean (95% confidence interval) dissociation constants were 38.9 um (33.2-44.6 um) and 36.9 um (26.7-47.1 um), and the numbers of binding sites were 1.36 (1.27-1.44) and 1.33 (1.19-1.47) in the monotherapy and polytherapy groups, respectively. No significant differences in the binding parameters of VPA to serum albumin were observed between the two groups. CONCLUSIONS: The steady-state serum albumin binding of VPA in Japanese patients with epilepsy is not affected by co administration of other antiepileptic drugs. These findings suggest that serum VPA concentration is stable at the steady state with regard to interaction by protein binding, even when other antiepileptic drugs with moderate-to-high binding properties are co-administered. PMID- 21635266 TI - Uncovering auditory evoked potentials from cochlear implant users with independent component analysis. AB - Auditory evoked potentials (AEPs) provide an objective measure of auditory cortical function, but AEPs from cochlear implant (CI) users are contaminated by an electrical artifact. Here, we investigated the effects of electrical artifact attenuation on AEP quality. The ability of independent component analysis (ICA) in attenuating the CI artifact while preserving the AEPs was evaluated. AEPs recovered from CI users were systematically correlated with age, demonstrating that individual differences were well preserved. CI users with high-quality AEPs were characterized by a significantly shorter duration of deafness. Finally, a simulation study revealed very high spatial correlations between original and recovered normal hearing AEPs (r>.95) that were previously contaminated with CI artifacts. The results confirm that after ICA, good quality AEPs can be recovered, facilitating the objective, noninvasive study of auditory cortex function in CI users. PMID- 21635267 TI - Prevalence of hymenoptera venom allergy and poor adherence to immunotherapy in Austria. PMID- 21635268 TI - The pharmacokinetics and metabolism of meloxicam in camels after intravenous administration. AB - The pharmacokinetics and metabolism of meloxicam was studied in camels (Camelus dromedarus) (n = 6) following intravenous (i.v.) administration of a dose of 0.6 mg.kg/body weight. The results obtained (mean +/- SD) were as follows: the terminal elimination half-life (t(1/2beta) ) was 40.2 +/- 16.8 h and total body clearance (Cl(T) ) was 1.94 +/- 0.66 mL.kg/h. The volume of distribution at steady state (V(SS)) was 92.8 +/- 13.7 mL/kg. One metabolite of meloxicam was tentatively identified as methylhydroxy meloxicam. Meloxicam and metabolite were excreted unconjugated in urine. Meloxicam could be detected in plasma 10 days following i.v. administration in camels using a sensitive liquid chromatography tandem mass spectrometry (LC/MS/MS) method. PMID- 21635269 TI - The application of delta18O and deltaD for understanding water pools and fluxes in a Typha marsh. AB - The delta18O and deltaD composition of water pools (leaf, root, standing water and soil water) and fluxes [transpiration (T), evaporation (E)] were used to understand ecohydrological processes in a managed Typha latifolia L. freshwater marsh. We observed isotopic steady-state T and deep rooting in Typha. The isotopic mass balance of marsh standing water showed that E accounted for 3% of the total water loss, T accounted for 17% and subsurface drainage (D) accounted for the majority (80%). There was a vertical gradient in water vapour content and isotopic composition within and above the canopy sufficient for constructing an isotopic mass balance of water vapour during some sampling periods. During these periods, the proportion of T in evapotranspiration (T/ET) was between 56 +/- 17% and 96 +/- 67%, and the estimated error was relatively high (>37%) because of non local, background sources in vapour. Independent estimates of T/ET using eddy covariance measurements yielded similar mean values during the Typha growing season. The various T/ET estimates agreed that T was the dominant source of marsh vapour loss in the growing season. The isotopic mass balance of water vapour yielded reasonable results, but the mass balance of standing water provided more definitive estimates of water losses. PMID- 21635270 TI - Is there an important role for reactive oxygen species and redox regulation during floral senescence? AB - Senescence is a highly regulated process terminating with programmed cell death (PCD). Floral senescence, and in particular petal senescence, forms an interesting model to study this process in that floral lifespan is species specific and linked to biological function. A feature of petal senescence is a rise in reactive oxygen species (ROS) and a change in redox balance. A key question is whether this is merely a consequence of de-regulation of antioxidant systems as cells enter PCD, or whether the rise in ROS may have a regulatory or signalling function. An important division in the physiology of floral senescence is between species in which ethylene is a key regulator, and those in which it appears not to perform an important regulatory role. Another important question we can therefore ask is whether the redox and ROS changes have the same significance in species with different physiologies. Transcriptomic studies in ethylene-sensitive and -insensitive species allow us to further determine whether changes in the activity of ROS-scavenging enzymes are transcriptionally regulated during floral senescence. Finally, it is important to assess how a signalling role for ROS or redox status would fit with known plant growth regulator (PGR) control of floral senescence. PMID- 21635271 TI - Regulation of RhSUC2, a sucrose transporter, is correlated with the light control of bud burst in Rosa sp. AB - In roses, light is a central environmental factor controlling bud break and involves a stimulation of sugar metabolism. Very little is known about the role of sucrose transporters in the bud break process and its regulation by light. In this study, we show that sugar promotes rose bud break and that bud break is accompanied by an import of sucrose. Radio-labelled sucrose accumulation is higher in buds exposed to light than to darkness and involves an active component. Several sucrose transporter (RhSUC1, 2, 3 and 4) transcripts are expressed in rose tissues, but RhSUC2 transcript level is the only one induced in buds exposed to light after removing the apical dominance. RhSUC2 is preferentially expressed in bursting buds and stems. Functional analyses in baker's yeast demonstrate that RhSUC2 encodes a sucrose/proton co-transporter with a K(m) value of 2.99 mm at pH 4.5 and shows typical features of sucrose symporters. We therefore propose that bud break photocontrol partly depends upon the modulation of sucrose import into buds by RhSUC2. PMID- 21635272 TI - The influence of ethnic origin on the skin photoageing: Nepalese study. AB - Ethnic and genetic differences modify skin structure and function. Skin photoageing is becoming one of the most studied cosmetological topics. However, there are relatively few data available to evaluate the effect of ethnic skin origin on the degree of photoageing, particularly among people inhabiting sunny and hot climate as in South Asia. Two hundred and forty participants, enrolled in the study conducted in Pokhara valley, Nepal, were classified into four different age categories viz. below 30, 30-50, 50-60 and above 60 years. Participants answered the questioner regarding their age, gender, ethnic origin, occupation, skin characteristics, lifestyle factors, type of cosmetic and skin care products used, drug treatment and personal skin condition evaluation. Skin hyperpigmentation and the degree of wrinkling, the two most important parameters related to photoageing, were particularly evaluated in two ethnic groups, namely Aryan-origin and Mongolian-origin participants. The study showed that Aryan origin ethnic skin was more wrinkled and darker than that of Mongolian. Male skin was found to be darker than the female skin, which might be the result of the use of sun-protective cosmetic products among women. Smoking was found to increase the degree of wrinkling; however, no differences related to ethnic origin in relation to smoking were observed. This study indicates the need to further evaluate the differences in skin response to daily exposure to strong sun among various ethnic-origin inhabitants. PMID- 21635273 TI - Motivated use of information about others: linking the 2 * 2 achievement goal model to social comparison propensities and processes. AB - The present research used correlational and experimental methods and two well established social comparison paradigms to integrate and extend prior research from the achievement goal and social comparison literatures. In Study 1, a general disposition to engage in social comparison was positively correlated with each type of goal in the 2 * 2 model of achievement goals, suggesting that the desire to seek out social comparison information is not exclusive to a particular type of achievement goal pursuit. In Study 2, when evaluating the specific direction of social comparison (upward or downward), the pursuit of performance approach, mastery-approach, and mastery-avoidance goals facilitated upward social comparison, and the pursuit of performance-avoidance goals prompted a shift away from upward comparison towards downward comparison. The present findings provide new insight to the emerging integration of achievement goals and social comparison. PMID- 21635274 TI - Effects of alcohols on recombinant adenylyl cyclase type 7 expressed in bacteria. AB - BACKGROUND: Our previous studies showed that ethanol enhanced the activity of adenylyl cyclase (AC) in an isoform-specific manner and that alcohol cutoff point of AC was isoform specific. Recently, we showed that 2,3-butanediol inhibited AC type 7 (AC7) activity in a stereoisomer-specific manner and that this inhibition was also AC isoform specific. These observations strongly suggest that a major target of alcohol action on cAMP signaling is AC. We hypothesized that alcohols exhibit their effect on AC activity by direct interaction with AC proteins. However, experimental systems employed in past studies such as intact cells and membrane preparations are too complex and do not allow us to unequivocally test this hypothesis. In attempt to bypass, these complications of the membrane-bound AC, we decided to study the effect of alcohols on AC recombinant proteins expressed in bacteria. METHODS: A recombinant AC, designated as AC7sol, consisting of the C(1a) and C(2) domains of the human AC7 was designed and expressed in bacteria. The activity of AC7sol was examined using lysate prepared from bacteria expressing AC7sol. RESULTS: The activity of AC7sol was stimulated by manganese or by the alpha subunit of G protein that stimulates AC (G(salpha) ). Forskolin by itself did not stimulate the activity of AC7sol. However, in the presence of activated G(salpha) , forskolin stimulated the activity of AC7sol. A series of n-alkanols including ethanol enhanced the manganese-stimulated activity of AC7sol. The alcohol cutoff point of AC7sol was pentanol. Ethanol and butanol increased V(max) and K(M) values of AC7sol. CONCLUSIONS: These results are consistent with our hypothesis and suggest that the enhancing effect of alcohols on AC activity is because of the increase in turnover number of AC. The current study demonstrates for the first time that the effect of alcohols requires only the C(1a) and C(2) domains of AC and no other domains of AC as well as no other mammalian proteins. PMID- 21635275 TI - Association of alcohol dehydrogenase genes with alcohol-related phenotypes in a Native American community sample. AB - BACKGROUND: Previous linkage studies, including a study of the Native American population described in the present report, have provided evidence for linkage of alcohol dependence and related traits to chromosome 4q near a cluster of alcohol dehydrogenase (ADH) genes, which encode enzymes of alcohol metabolism. METHODS: The present study tested for associations between alcohol dependence and related traits and 22 single-nucleotide polymorphisms (SNPs) spanning the 7 ADH genes. Participants included 586 adult men and women recruited from 8 contiguous Native American reservations. A structured interview was used to assess DSM-III-R alcohol dependence criteria as well as a set of severe alcohol misuse symptoms and alcohol withdrawal symptoms. RESULTS: No evidence for association with the alcohol dependence diagnosis was observed, but an SNP in exon 9 of ADH1B (rs2066702; ADH1B*3) and an SNP at the 5' end of ADH4 (rs3762894) showed significant evidence of association with the presence of withdrawal symptoms (p = 0.0018 and 0.0012, respectively). Further, a haplotype analysis of these 2 SNPs suggested that the haplotypes containing either of the minor alleles were protective against alcohol withdrawal relative to the ancestral haplotype (p = 0.000006). CONCLUSIONS: These results suggest that variants in the ADH1B and ADH4 genes may be protective against the development of some symptoms associated with alcohol dependence. PMID- 21635276 TI - Structure and activity of lacustrine sediment bacteria involved in nutrient and iron cycles. AB - Knowledge of the bacterial community structure in sediments is essential to better design restoration strategies for eutrophied lakes. In this regard, the aim of this study was to quantify the abundance and activity of bacteria involved in nutrient and iron cycling in sediments from four Azorean lakes with distinct trophic states (Verde, Azul, Furnas and Fogo). Inferred from quantitative PCR, bacteria performing anaerobic ammonia oxidation were the most abundant in the eutrophic lakes Verde, Azul and Furnas (4.5-16.6%), followed by nitrifying bacteria (0.8-13.0%), denitrifying bacteria (DNB) (0.5-6.8%), iron-reducing bacteria (0.2-1.4%) and phosphorus-accumulating organisms (<0.3%). In contrast, DNB dominated sediments from the oligo-mesotrophic lake Fogo (8.8%). Activity assays suggested that bacteria performing ammonia oxidation (aerobic and anaerobic), nitrite oxidation, heterothrophic nitrate reduction, iron reduction and biological phosphorus storage/release were present and active in all Azorean lake sediments. The present work also suggested that the activity of DNB might contribute to the release of phosphorus from sediments. PMID- 21635277 TI - Very low birth weight survivors have reduced peak bone mass and reduced insulin sensitivity. AB - CONTEXT: Increasing numbers of very low birth weight (VLBW) infants are surviving into adulthood because of improvements in neonatal intensive care. Adverse events in early life can have long-term effects through reprogramming of metabolic systems. OBJECTIVE: To determine whether young adult VLBW survivors have abnormalities of skeletal development or endocrine function. DESIGN: Cross sectional, observational, case-control study. PARTICIPANTS: Thirty-seven VLBW subjects and 27 healthy controls at peak bone mass (mean age 23). MEASUREMENTS: Differences between cases and controls in body size, body composition, bone mass and bone geometry [assessed by dual-energy X-ray absorptiometry (DXA), hip structure analysis and peripheral quantitative computed tomography (pQCT)], bone turnover [urine N-terminal telopeptide of type I collagen (NTX), serum C-terminal telopeptide of type I collagen (CTX)], aminoterminal propeptide of type I procollagen (PINP) and bone alkaline phosphatase), hormones (sex steroids, IGF-1, PTH and 25-OH vitamin D) and insulin sensitivity (HOMA-IR and oral glucose tolerance testing). RESULTS: VLBW subjects had lower bone density at the lumbar spine (5.7%) and femoral neck (8.6%), which persisted after correction for bone size by the estimation of volumetric density (bone mineral apparent density). Urine NTX was higher in VLBW subjects than in controls, but there were no significant differences in other bone turnover markers. VLBW survivors had lower insulin sensitivity (mean INS-30 controls = 57.0, VLBW subjects = 94.3, P < 0.01), but there were no differences in whole body fat mass or truncal fat mass between VLBW subjects and controls. CONCLUSIONS: Young adult VLBW survivors have reduced bone density for their bone size and reduced insulin sensitivity, which may have significant implications for their risk of fracture and diabetes in later life. PMID- 21635278 TI - Surgical therapy of peri-implantitis lesions by means of a bovine-derived xenograft: comparative results of a prospective study on two different implant surfaces. AB - OBJECTIVES: The aim of this prospective study was to evaluate a regenerative surgical treatment modality for peri-implantitis lesions on two different implant surfaces. MATERIALS AND METHODS: Twenty-six patients with one crater-like defect, around either TPS (Control) or SLA (Test) dental implants, with a probing depth (PD) >=6 mm and no implant mobility, were included. The implant surface was mechanically debrided and treated using a 24% EDTA gel and a 1% chlorhexidine gel. The bone defect was filled with a bovine-derived xenograft (BDX) and the flap was sutured around the non-submerged implant. RESULTS: One-year follow-up demonstrated clinical and radiographic improvements. PDs were significantly reduced by 2.1+/-1.2 mm in the Control implants and by 3.4+/-1.7 mm in the Test implants. Complete defect fill was never found around Controls, while it occurred in three out of 12 Test implants. Bleeding on probing decreased from 91.1+/-12.4% (Control) and 75.0+/-30.2% (Test) to 57.1+/-38.5% (p=0.004) and 14.6+/-16.7% (p=0.003), respectively. Several deep pockets (>=6 mm) were still present after surgical therapy around Controls. CONCLUSIONS: Surface characteristics may have an impact on the clinical outcome following surgical debridement, disinfection of the contaminated surfaces and grafting with BDX. Complete fill of the bony defect seems not to be a predictable result. PMID- 21635279 TI - Fourteen-year outcomes of coronally advanced flap for root coverage: follow-up from a randomized trial. AB - TRIAL DESIGN: This long-term 14-year-randomized split-mouth study aimed at evaluating (1) the outcomes of two different methods of root surface modifications (root surface polishing versus root planing) used in combination with a coronally advanced flap (CAF) and (2) the long-term results of CAF performed for the treatment of single gingival recessions. METHODS: Ten patients with similar bilateral recessions >=2 mm were selected for a split-mouth randomized design study. Exposed root surfaces were assigned to receive polishing (test sites) or root planing (control sites). A multilevel model was used to analyse data at 3 months, 1, 5 and 14 years. RESULTS: One patient dropped out after 1 year. At 14 years, recession depth (Rec) was 0.9 (1.2) mm for the test sites and 0.9 (0.9) mm for the control sites. The interaction between treatment and keratinized tissue was significant (p=0.0035). Rec increased slightly over time (p=0.0006) in both the groups. CONCLUSIONS: This study shows that during a long-term follow-up, gingival recession recurred in 39% of the treated sites following the CAF procedure. PMID- 21635280 TI - Periodontal disease as a risk for dental implant failure over time: a long-term historical cohort study. AB - OBJECTIVES: To evaluate the long-term survival rates of dental implants according to the patient's periodontal status, as well as to estimate if the effect of periodontal status regarding implant failure is constant throughout the long-term follow-up. MATERIALS AND METHODS: This was a historical prospective cohort study design of all consecutive patients operated from 1996 to 2006 at a periodontal clinic. The cohort consisted of 736 patients, with a total of 2336 dental implants. An extended Cox proportional hazards model, which includes interaction terms between survival time and variables of interest, was used. RESULTS: Patients' mean (SD) age was 51.13 (12.35). The follow-up time was up to 144 months, with a mean (SD) of 54.4 (35.6) months. The overall implant raw survival rate was 95.9%. The Kaplan-Meier estimates for the cumulative survival rate (CSR) at 108 months were 0.96 and 0.95 for implants inserted into healthy and moderate chronic periodontal patients, respectively. The CSR declined to 0.88 at 108 months for the severe periodontitis group. The extended Cox model revealed that severe chronic status turned out to be a significant risk factor for implant failure after 50 months of follow-up [hazard ratio (HR)=8.06; p<0.01]. The extended Cox model for smoking indicates a near-significant effect after 50 months (HR=2.76; p=0.061). CONCLUSIONS: Periodontal status and smoking are significant risk factors for late implant failures. The HR for periodontal and smoking status are not constant throughout the follow-up period. PMID- 21635281 TI - Living with severe mental illness: perception of sickness. AB - AIM: This article is a report of a study on how participation in photo groups influenced the perception of the impact of sickness on daily life. BACKGROUND: Living with severe mental illness can have a strong impact on daily life and the experience of self. In combination with self-stigmatization, this can lead to a sense of being no more than a patient. METHOD: A mixed-methods design was used to measure the perception of sickness impact. Consumers of psychiatric services participated in photography groups that aimed at integration of illness and developing new goals in life. These groups were conducted by nurses and data were collected between the years 2005 and 2009. The Sickness Impact Profile (SIP) questionnaires were completed by the respondents before and after intervention. The SIP was used to differentiate between respondents who perceived less impact of illness on their daily lives after following a photogroup and those who did not perceive so or remained the same. A sample of these three groups was used to complement the quantitative findings with in-depth interviews with consumers and mentor nurses. RESULTS: Findings indicated absence of important changes in the sickness impact scores except in the domain of mobility. Consumers did, however, show signs of progress in terms of increasing openness, understanding of their feelings and situation and abilities to cope with them. CONCLUSION: Participating in photo groups can help patients get along with their life and make it more bearable. No signs of demoralization and self-stigmatization were found. PMID- 21635282 TI - A confirmatory study of violence risk assessment tool (M55) and demographic predictors of patient violence. AB - AIM: The aims of this study were to confirm the usefulness of Violence Risk Assessment Tool (M55) for prospectively identifying violent patients in medical surgical units and to explore predictors of Code-55 activation for violent behaviour or physical attack. BACKGROUND: A previous retrospective case controlled study claimed moderate sensitivity and high specificity of the M55 in identifying potentially violent patients. However, the usefulness of M55 needs to be confirmed in a prospective study before wide adoption. METHODS: A prospective cohort study of patients in medical-surgical units of a hospital in southern California was conducted from August 2009 to December 2009. A checklist containing the M55 was completed on admission. Following a violent event or just prior to discharge, the violent event outcome section of the data collection tool was completed. The sensitivity and specificity of M55 for identifying the violent patients were calculated using the original M55 flagging criteria. RESULTS: Of 2063 patients, 32 (1.6%) had a violent event of Code-55 activation (12), physical attack (26) or both (6). The M55 showed a low sensitivity (41%) and acceptable specificity (99%) for prospectively identifying violent patients. The multivariate logistic regression model indicated higher odds of violent events for patients >=70 years of age [odds ratio (OR) = 2.3; 95% confidence interval (CI): 0.9-5.7], males (OR = 2.7; 95% CI: 1.2-6.1) or Caucasian (OR = 3.4; 95% CI: 0.8-14.7). CONCLUSION: The M55 does not appear to be useful for prospectively identifying violent patients in medical-surgical units because of low sensitivity. Older, male or Caucasian patients had higher odds of becoming violent. PMID- 21635283 TI - Efficacy of penguin cap as scalp cooling system for prevention of alopecia in patients undergoing chemotherapy. AB - AIMS: The present study examines the efficacy of a scalp cooling system to prevent chemotherapy-induced alopecia in people with cancer. BACKGROUND: Alopecia has been established as the most distressing aspect of chemotherapy to such an extent that some patients even refrain from cytotoxic treatment. METHODS: Using a quasi-experimental approach in 2007, a total number of 63 patients aged 16-59 years (mean 35 +/- 5.5 years) participated in the study and were treated with an intravenous regimen of chemotherapy drug. Thirty-one patients accepting the scalp cooling system were compared for alopecia with 32 participants who refused it. Hair loss in the participants was evaluated by nurses using World Health Organization criteria at each cycle of chemotherapy. RESULTS: Using chi-square test differences between the two groups were statistically significant at cycles 2-6 (P < 0.05). Twenty-four participants (77.4%) of the study group were assessed as first and second grade of alopecia in the second cycle, whereas seven participants (22.6%) of this group exhibited third and fourth grade of alopecia. However, in the control group at the same cycle, 19 participants (61.3%) were assessed with third and fourth grade of alopecia. In the Sixth cycle 15 participants (50%) of the study group and 24 participants (75%) of the control group demonstrated third and fourth grade alopecia. CONCLUSION: A penguin cap can be an effective method to avoid alopecia in patients receiving chemotherapy drugs. However, it is important to bear in mind that because of individual characteristics, liver function and drug regimens it is not successful in all cases. PMID- 21635284 TI - Self-care of school-age children with diabetes: an integrative review. AB - AIM: This paper is a report of an integrative review of findings from empirical studies on self-care in school-age children with type 1 diabetes. The purpose is to generate insight into opportunities to develop empowering patient education. BACKGROUND: Managing diabetes is demanding and requires parental involvement in care. Good self-care forms the basis for diabetes management and self-care patterns are established at school age, but how and to what extent school-age children increase their self-care capabilities is unclear. DATA SOURCES: A search for studies from 1998 to 2010 focusing on self-care in school-age children with diabetes was conducted through electronic databases. REVIEW METHODS: Using integrative methods, quantitative and qualitative papers surveyed were analysed separately, but the themes that arose were combined at the end of the analysis. FINDINGS: Self-care is formed in a learning process involving the objectives of normality, being able to cope and independence. The content of self-care is a combination of knowledge and skills. Children have the technical skill, but they need their parents to participate in the care and share responsibility for it. The factors related to self-care comprised the characteristics of the child; the nature of the illness and care; and support from the parents, school environment, peers and healthcare team. CONCLUSION: A balance between diabetes care requirements and a child's maturity should be found. Nurses must adopt an empowering manner of education and recognize and assess a child's readiness to learn diabetes care and bear responsibility for it. Nurses must also help parents and other adults to gradually shift the responsibility to the children. PMID- 21635285 TI - Combat stressors and post-traumatic stress in deployed military healthcare professionals: an integrative review. AB - BACKGROUND: While there has been a growing body of literature on the impact of combat stressors and post-traumatic stress on military service members involved in current conflicts, there has been little available information that directly examines the impact of these on healthcare providers. AIMS: Aims for this integrative review included: (1) identifying exposures, experiences and other factors influencing stress responses in military healthcare providers previously engaged in a war effort and (2) describing the incidence of post-traumatic stress and related mental health problems in this population. REVIEW METHODS: Using Cooper's integrative review method, relevant documents were collected and analysed using content categories and a coding scheme to assist with identifying and recording data for units of analysis. DATA SOURCES: Literature searches (including all years to present) were conducted using keywords for stress reaction, for healthcare provider and for military war effort involvement. Literature was obtained using the Cumulative Index to Nursing and Allied Health Literature, the National Library of Medicine and the American Psychological Association databases. RESULTS: Evidence suggests that similar to military combatants, military healthcare provider exposure to life-threatening situations will increase the probability of adverse psychological disorders following these traumatic experiences. The presence of a strong sense of meaning and purpose, within a supportive environment appear to help mediate the impact of these dangerous and stressful events. CONCLUSION: Results of this review and other supporting literature indicate the need for a systematic approach to studying combat stress and post-traumatic stress in deployed healthcare providers. PMID- 21635286 TI - A tribute to larry erlbaum. PMID- 21635287 TI - When are tutorial dialogues more effective than reading? AB - It is often assumed that engaging in a one-on-one dialogue with a tutor is more effective than listening to a lecture or reading a text. Although earlier experiments have not always supported this hypothesis, this may be due in part to allowing the tutors to cover different content than the noninteractive instruction. In 7 experiments, we tested the interaction hypothesis under the constraint that (a) all students covered the same content during instruction, (b) the task domain was qualitative physics, (c) the instruction was in natural language as opposed to mathematical or other formal languages, and (d) the instruction conformed with a widely observed pattern in human tutoring: Graesser, Person, and Magliano's 5-step frame. In the experiments, we compared 2 kinds of human tutoring (spoken and computer mediated) with 2 kinds of natural-language based computer tutoring (Why2-Atlas and Why2-AutoTutor) and 3 control conditions that involved studying texts. The results depended on whether the students' preparation matched the content of the instruction. When novices (students who had not taken college physics) studied content that was written for intermediates (students who had taken college physics), then tutorial dialogue was reliably more beneficial than less interactive instruction, with large effect sizes. When novices studied material written for novices or intermediates studied material written for intermediates, then tutorial dialogue was not reliably more effective than the text-based control conditions. PMID- 21635288 TI - Speed, accuracy, and serial order in sequence production. AB - The production of complex sequences like music or speech requires the rapid and temporally precise production of events (e.g., notes and chords), often at fast rates. Memory retrieval in these circumstances may rely on the simultaneous activation of both the current event and the surrounding context (Lashley, 1951). We describe an extension to a model of incremental retrieval in sequence production (Palmer & Pfordresher, 2003) that incorporates this logic to predict overall error rates and speed-accuracy trade-offs, as well as types of serial ordering errors. The model-assumes that retrieval of the current event is influenced by activations of surrounding events. Activations of surrounding events increase over time, such that both the accessibility of distant events and overall accuracy increases at slower production rates. The model's predictions were tested in an experiment in which pianists performed unfamiliar music at 8 different tempi. Model fits to speed-accuracy data and to serial ordering errors support model predictions. Parameter fits to individual data further suggest that working memory contributes to the retrieval of serial order and overall accuracy is influenced in addition by motor dexterity and domain-specific skill. PMID- 21635289 TI - Explaining color term typology with an evolutionary model. AB - An expression-induction model was used to simulate the evolution of basic color terms to test Berlin and Kay's (1969) hypothesis that the typological patterns observed in basic color term systems are produced by a process of cultural evolution under the influence of biases resulting from the special properties of universal focal colors. Ten agents were simulated, each of which could learn color term denotations by generalizing from examples using Bayesian inference, and for which universal focal red, yellow, green, and blue were especially salient, but unevenly spaced in the perceptual color space. Conversations between these agents, in which agents would learn from one another, were simulated over several generations, and the languages emerging at the end of each simulation were investigated. The proportion of color terms of each type correlated closely with the equivalent frequencies found in the World Color Survey, and most of the emergent languages could be placed on one of the evolutionary trajectories proposed by Kay and Maffi (1999). The simulation therefore demonstrates how typological patterns can emerge as a result of learning biases acting over a period of time. PMID- 21635290 TI - The dynamics of lexical competition during spoken word recognition. AB - The sounds that make up spoken words are heard in a series and must be mapped rapidly onto words in memory because their elements, unlike those of visual words, cannot simultaneously exist or persist in time. Although theories agree that the dynamics of spoken word recognition are important, they differ in how they treat the nature of the competitor set-precisely which words are activated as an auditory word form unfolds in real time. This study used eye tracking to measure the impact over time of word frequency and 2 partially overlapping competitor set definitions: onset density and neighborhood density. Time course measures revealed early and continuous effects of frequency (facilitatory) and on set based similarity (inhibitory). Neighborhood density appears to have early facilitatory effects and late inhibitory effects. The late inhibitory effects are due to differences in the temporal distribution of similarity within neighborhoods. The early facilitatory effects are due to subphonemic cues that inform the listener about word length before the entire word is heard. The results support a new conception of lexical competition neighborhoods in which recognition occurs against a background of activated competitors that changes over time based on fine-grained goodness-of-fit and competition dynamics. PMID- 21635291 TI - Formal distinctiveness of high- and low-imageability nouns: analyses and theoretical implications. AB - Words associated with perceptually salient, highly imageable concepts are learned earlier in life, more accurately recalled, and more rapidly named than abstract words (R. W. Brown, 1976; Walker & Hulme, 1999). Theories accounting for this concreteness effect have focused exclusively on semantic properties of word referents. A novel possibility is that word structure may also contribute to the effect. We report a corpus-based analysis of the phonological and morphological structures of a large set of nouns with imageability ratings (N = 2,023). High- and low-imageability nouns differed by length, etymology, prosody, affixation, phonological neighborhood density, and rates of consonant clustering. On average, nouns denoting abstract concepts were longer, more derivationally complex, and emerged in English from a different distribution of languages than did concrete nouns. We address implications for interactivity of word form and meaning as pertain to theories of word concreteness, lexical acquisition, and word processing. PMID- 21635292 TI - The effect of information overlap on communication effectiveness. AB - It makes sense that the more information people share, the better they communicate. To evaluate the effect of knowledge overlap on the effectiveness of communication, participants played a communication game where the "director" identified objects to the "addressee". Pairs either shared information about most objects' names (high overlap), or about the minority of objects' names (low overlap). We found that high-overlap directors tended to use more names than low overlap directors. High overlap directors also used more names with objects whose names only they knew, thereby confusing their addressees more often than low overlap directors. We conclude that while sharing more knowledge can be beneficial to communication overall, it can cause communication to be locally ineffective. Sharing more information reduces communication effectiveness precisely when there is an opportunity to inform-when people communicate information only they themselves know. PMID- 21635293 TI - Switching between sensory and affective systems incurs processing costs. AB - Recent models of the conceptual system hold that concepts are grounded in simulations of actual experiences with instances of those concepts in sensory motor systems (e.g., Barsalou, 1999, 2003; Solomon & Barsalou, 2001). Studies supportive of such a viewhave shown that verifying a property of a concept in one modality, and then switching to verify a property of a different concept in a different modality generates temporal processing costs similar to the cost of switching modalities in perception. In addition to non-emotional concepts, the present experiment investigated switching costs in verifying properties of positive and negative (emotional) concepts. Properties of emotional concepts were taken from vision, audition, and the affective system. Parallel to switching costs in neutral concepts, the study showed that for positive and negative concepts, verifying properties from different modalities produced processing costs such that reaction times were longer and error rates were higher. Importantly, this effect was observed when switching from the affective system to sensory modalities, and vice-versa. These results support the embodied cognition view of emotion in humans. PMID- 21635294 TI - The cognition of engineering design-an opportunity of impact. PMID- 21635295 TI - Situated language understanding as filtering perceived affordances. AB - We introduce a computational theory of situated language understanding in which the meaning of words and utterances depends on the physical environment and the goals and plans of communication partners. According to the theory, concepts that ground linguistic meaning are neither internal nor external to language users, but instead span the objective-subjective boundary. To model the possible interactions between subject and object, the theory relies on the notion of perceived affordances: structured units of interaction that can be used for prediction at multiple levels of abstraction. Language understanding is treated as a process of filtering perceived affordances. The theory accounts for many aspects of the situated nature of human language use and provides a unified solution to a number of demands on any theory of language understanding including conceptual combination, prototypicality effects, and the generative nature of lexical items. To support the theory, we describe an implemented system that understands verbal commands situated in a virtual gaming environment. The implementation uses probabilistic hierarchical plan recognition to generate perceived affordances. The system has been evaluated on its ability to correctly interpret free-form spontaneous verbal commands recorded from unrehearsed game play between human players. The system is able to "step into the shoes" of human players and correctly respond to a broad range of verbal commands in which linguistic meaning depends on social and physical context. We quantitatively compare the system's predictions in response to direct player commands with the actions taken by human players and show generalization to unseen data across a range of situations and verbal constructions. PMID- 21635296 TI - Combining versus analyzing multiple causes: how domain assumptions and task context affect integration rules. AB - In everyday life, people typically observe fragments of causal networks. From this knowledge, people infer how novel combinations of causes they may never have observed together might behave. I report on 4 experiments that address the question of how people intuitively integrate multiple causes to predict a continuously varying effect. Most theories of causal induction in psychology and statistics assume a bias toward linearity and additivity. In contrast, these experiments show that people are sensitive to cues biasing various integration rules. Causes that refer to intensive quantities (e.g., taste) or to preferences (e.g., liking) bias people toward averaging the causal influences, whereas extensive quantities (e.g., strength of a drug) lead to a tendency to add. However, the knowledge underlying these processes is fallible and unstable. Therefore, people are easily influenced by additional task-related context factors. These additional factors include the way data are presented, the difficulty of the inference task, and transfer from previous tasks. The results of the experiments provide evidence for causal model and related theories, which postulate that domain-general representations of causal knowledge are influenced by abstract domain knowledge, data-driven task factors, and processing difficulty. PMID- 21635297 TI - Internalism, active externalism, and nonconceptual content: the ins and outs of cognition. AB - Active externalism (also known as the extended mind hypothesis) says that we use objects and situations in the world as external memory stores that we consult as needs dictate. This gives us economies of storage: We do not need to remember that Bill has blue eyes and wavy hair if we can acquire this information by looking at Bill. I argue for a corollary to this position, which I call 'internalism.' Internalism says we can acquire knowledge on a need-to-know basis by consulting portable, inner analogues of the world. This, however, leads to a dilemma. If the knowledge was stored in memory, it is difficult to see how we could have acquired it by consulting inner analogues, for it would seem that we knew it already. Moreover, if it was stored in memory, we lose the economies of storage that provide much of the rationale for external memory stores and hence for their inner analogues. If, on the other hand, the knowledge was not stored in memory, it is difficult to see how we could have acquired it by consulting inner analogues, for it was not there to be acquired. I propose a solution to this problem that turns on the concept of nonconceptual content, and I relate the solution to Stephen Kosslyn's (1994) architecture of perception and visual mental imagery. Viewed in a broader context, the solution shows that the world leaks into the mind, as well as mind leaking into the world. PMID- 21635298 TI - Graphical language games: interactional constraints on representational form. AB - The emergence of shared symbol systems is considered to be a pivotal moment in human evolution and human development. These changes are normally explained by reference to changes in people's internal cognitive processes. We present 2 experiments which provide evidence that changes in the external, collaborative processes that people use to communicate can also affect the structure and organization of symbol systems independently of cognitive change. We propose that mutual-modifiability-opportunities for people to edit or manipulate each other's contributions-is a key constraint on the emergence of complex symbol systems. We discuss the implications for models of language development and the origins of compositionality. PMID- 21635299 TI - Modeling the Developmental Patterning of Finiteness Marking in English, Dutch, German, and Spanish Using MOSAIC. AB - In this study, we apply MOSAIC (model of syntax acquisition in children) to the simulation of the developmental patterning of children's optional infinitive (OI) errors in 4 languages: English, Dutch, German, and Spanish. MOSAIC, which has already simulated this phenomenon in Dutch and English, now implements a learning mechanism that better reflects the theoretical assumptions underlying it, as well as a chunking mechanism that results in frequent phrases being treated as 1 unit. Using 1, identical model that learns from child-directed speech, we obtain a close quantitative fit to the data from all 4 languages despite there being considerable cross-linguistic and developmental variation in the OI phenomenon. MOSAIC successfully simulates the difference between Spanish (a pro-drop language in which OI errors are virtually absent) and obligatory subject languages that do display the OI phenomenon. It also highlights differences in the OI phenomenon across German and Dutch, 2 closely related languages whose grammar is virtually identical with respect to the relation between finiteness and verb placement. Taken together, these results suggest that (a) cross-linguistic differences in the rates at which children produce OIs are graded, quantitative differences that closely reflect the statistical properties of the input they are exposed to and (b) theories of syntax acquisition need to consider more closely the role of input characteristics as determinants of quantitative differences in the cross linguistic patterning of phenomena in language acquisition. PMID- 21635300 TI - ERP Evidence for the Rapid Assignment of an (Appropriate) Antecedent to PRO. AB - Event-related brain potentials were recorded while subjects listened to sentences containing a controlled infinitival complement. Subject and object control items were used, both with 2 potential antecedents in the upper clause. Half of the sentences had a gender agreement violation between the null subject of the infinitival complement and an adjective predicated of it. The rapid detection of this anomaly would indicate that the parser had established the coreference relation between the null subject and an antecedent, and that the processor had rapidly consulted verb control information to select the proper antecedent of the null subject. The results showed that for both subject and object control items ungrammatical adjectives elicited a P600 effect. These data imply that the processor has coindexed the null subject with an antecedent, and that the antecedent has been selected on the basis of control information. These results are compatible with parsing models that emphasize the rapid influence of verb specific information on sentence processing. PMID- 21635301 TI - Typicality, graded membership, and vagueness. AB - This paper addresses theoretical problems arising from the vagueness of language terms, and intuitions of the vagueness of the concepts to which they refer. It is argued that the central intuitions of prototype theory are sufficient to account for both typicality phenomena and psychological intuitions about degrees of membership in vaguely defined classes. The first section explains the importance of the relation between degrees of membership and typicality (or goodness of example) in conceptual categorization. The second and third section address arguments advanced by Osherson and Smith (1997), and Kamp and Partee (1995), that the two notions of degree of membership and typicality must relate to fundamentally different aspects of conceptual representations. A version of prototype theory-the Threshold Model-is proposed to counter these arguments and three possible solutions to the problems of logical selfcontradiction and tautology for vague categorizations are outlined. In the final section graded membership is related to the social construction of conceptual boundaries maintained through language use. PMID- 21635302 TI - Language-relative construal of individuation constrained by universal ontology: revisiting language universals and linguistic relativity. AB - Objects and substances bear fundamentally different ontologies. In this article, we examine the relations between language, the ontological distinction with respect to individuation, and the world. Specifically, in cross-linguistic developmental studies that follow Imai and Gentner (1997), we examine the question of whether language influences our thought in different forms, like (1) whether the language-specific construal of entities found in a word extension context (Imai & Gentner, 1997) is also found in a nonlinguistic classification context; (2) whether the presence of labels per se, independent of the count-mass syntax, fosters ontology-based classification; (3) in what way, if at all, the count-mass syntax that accompanies a label changes English speakers' default construal of a given entity? On the basis of the results, we argue that the ontological distinction concerning individuation is universally shared and functions as a constraint on early learning of words. At the same time, language influences one's construal of entities cross-lingistically and developmentally, and causes a temporary change of construal within a single language. We provide a detailed discussion of how each of these three ways language may affect the construal of entities, and discuss how our universally possessed knowledge interacts with language both within a single language and in cross-linguistic context. PMID- 21635303 TI - Contextualizing counterintuitiveness: how context affects comprehension and memorability of counterintuitive concepts. AB - A number of anthropologists have argued that religious concepts are minimally counterintuitive and that this gives them mnemic advantages. This paper addresses the question of why people have the memory architecture that results in such concepts being more memorable than other types of concepts by pointing out the benefits of a memory structure that leads to better recall for minimally counterintuitive concepts and by showing how such benefits emerge in the real time processing of comprehending narratives such as folk tales. This model suggests that memorability is not an inherent property of a concept; rather it is a property of the concept, the context in which the concept is presented, and the background knowledge that the comprehendor possesses about the concept. The model predicts how memorability of a concept should change if the context containing the concept were changed. The paper also presents the results of experiments carried out to test these predictions. PMID- 21635304 TI - Language evolution by iterated learning with bayesian agents. AB - Languages are transmitted from person to person and generation to generation via a process of iterated learning: people learn a language from other people who once learned that language themselves. We analyze the consequences of iterated learning for learning algorithms based on the principles of Bayesian inference, assuming that learners compute a posterior distribution over languages by combining a prior (representing their inductive biases) with the evidence provided by linguistic data. We show that when learners sample languages from this posterior distribution, iterated learning converges to a distribution over languages that is determined entirely by the prior. Under these conditions, iterated learning is a form of Gibbs sampling, a widely-used Markov chain Monte Carlo algorithm. The consequences of iterated learning are more complicated when learners choose the language with maximum posterior probability, being affected by both the prior of the learners and the amount of information transmitted between generations. We show that in this case, iterated learning corresponds to another statistical inference algorithm, a variant of the expectation maximization (EM) algorithm. These results clarify the role of iterated learning in explanations of linguistic universals and provide a formal connection between constraints on language acquisition and the languages that come to be spoken, suggesting that information transmitted via iterated learning will ultimately come to mirror the minds of the learners. PMID- 21635305 TI - The role of prior experience in language acquisition. AB - Learners exposed to an artificial language recognize its abstract structural regularities when instantiated in a novel vocabulary (e.g., Gomez, Gerken, & Schvaneveldt, 2000; Tunney & Altmann, 2001). We asked whether such sensitivity accelerates subsequent learning, and enables acquisition of more complex structure. In Experiment 1, pre-exposure to a category-induction language of the form aX bY sped subsequent learning when the language is instantiated in a different vocabulary. In Experiment 2, while naive learners did not acquire an acX bcY language, in which aX and bY co-occurrence regularities were separated by a c-element, prior experience with an aX bY language provided some benefit. In Experiment 3 we replicated this finding with a 24-hour delay between learning phases, and controlled for prior experience with the aX bY language's prosodic and phonological characteristics. These findings suggest that learners, and the structure they can acquire, change as a function of experience. PMID- 21635306 TI - The role of falsification in the development of cognitive architectures: insights from a lakatosian analysis. AB - It has been suggested that the enterprise of developing mechanistic theories of the human cognitive architecture is flawed because the theories produced are not directly falsifiable. Newell attempted to sidestep this criticism by arguing for a Lakatosian model of scientific progress in which cognitive architectures should be understood as theories that develop over time. However, Newell's own candidate cognitive architecture adhered only loosely to Lakatosian principles. This paper reconsiders the role of falsification and the potential utility of Lakatosian principles in the development of cognitive architectures. It is argued that a lack of direct falsifiability need not undermine the scientific development of a cognitive architecture if broadly Lakatosian principles are adopted. Moreover, it is demonstrated that the Lakatosian concepts of positive and negative heuristics for theory development and of general heuristic power offer methods for guiding the development of an architecture and for evaluating the contribution and potential of an architecture's research program. PMID- 21635307 TI - What makes people revise their beliefs following contradictory anecdotal evidence?: the role of systemic variability and direct experience. AB - The extent to which belief revision is affected by systematic variability and direct experience of a conditional (if A then B) relation was examined in two studies. The first used a computer generated apparatus. This presented two rows of 5 objects. Pressing one of the top objects resulted in one of the bottom objects being lit up. The 139 adult participants were given one of two levels of experience (5 or 15 trials) and one of two types of apparatus. One of these was completely uniform, while the other had an element that randomly alternated in its result. Following the testing of the apparatus, participants were asked to rate their certainty of the action of the middle element, which was always uniform (the AB belief). Then they were told of an observation inconsistent with this belief. Participants were then asked whether they considered the AB belief or the anecdotal observation to be more believable. Results showed that increased experience decreased the tendency to reject the AB belief, when the apparatus did not have any randomness. However, the presence of a single element showing random variation in the system strongly increased rejection of this belief. A second study looked at the effect of a single random element on a mechanical system as well as an electronic system using graphical representations. This confirmed the generality of the effect of randomness on belief revision, and provided support for the effects of embedding a belief into a system of relations. These results provide some insight into the complex factors that determine belief revision. PMID- 21635308 TI - Spatial visualization in physics problem solving. AB - Three studies were conducted to examine the relation of spatial visualization to solving kinematics problems that involved either predicting the two-dimensional motion of an object, translating from one frame of reference to another, or interpreting kinematics graphs. In Study 1, 60 physics-naive students were administered kinematics problems and spatial visualization ability tests. In Study 2, 17 (8 high- and 9 low-spatial ability) additional students completed think-aloud protocols while they solved the kinematics problems. In Study 3, the eye movements of fifteen (9 high- and 6 low-spatial ability) students were recorded while the students solved kinematics problems. In contrast to high spatial students, most low-spatial students did not combine two motion vectors, were unable to switch frames of reference, and tended to interpret graphs literally. The results of the study suggest an important relationship between spatial visualization ability and solving kinematics problems with multiple spatial parameters. PMID- 21635309 TI - Dynamic self-organization and early lexical development in children. AB - In this study we present a self-organizing connectionist model of early lexical development. We call this model DevLex-II, based on the earlier DevLex model. DevLex-II can simulate a variety of empirical patterns in children's acquisition of words. These include a clear vocabulary spurt, effects of word frequency and length on age of acquisition, and individual differences as a function of phonological short-term memory and associative capacity. Further results from lesioned models indicate developmental plasticity in the network's recovery from damage, in a non-monotonic fashion. We attribute the network's abilities in accounting for lexical development to interactive dynamics in the learning process. In particular, variations displayed by the model in the rate and size of early vocabulary development are modulated by (a) input characteristics, such as word frequency and word length, (b) consolidation of lexical-semantic representation, meaning-form association, and phonological short-term memory, and (c) delayed processes due to interactions among timing, severity, and recoverability of lesion. Together, DevLex and DevLex-II provide an accurate computational account of early lexical development. PMID- 21635310 TI - A computational model of event segmentation from perceptual prediction. AB - People tend to perceive ongoing continuous activity as series of discrete events. This partitioning of continuous activity may occur, in part, because events correspond to dynamic patterns that have recurred across different contexts. Recurring patterns may lead to reliable sequential dependencies in observers' experiences, which then can be used to guide perception. The current set of simulations investigated whether this statistical structure within events can be used 1) to develop stable internal representations that facilitate perception and 2) to learn when to update such representations in a self-organizing manner. These simulations demonstrate that experience with recurring patterns enables a system to accurately predict upcoming stimuli within an event, to identify boundaries between such events based on transient increases in prediction error, and to use such boundaries to improve prediction about subsequent activities. PMID- 21635311 TI - Semantic boost on episodic associations: an empirically-based computational model. AB - Words become associated following repeated co-occurrence episodes. This process might be further determined by the semantic characteristics of the words. The present study focused on how semantic and episodic factors interact in incidental formation of word associations. First, we found that human participants associate semantically related words more easily than unrelated words; this advantage increased linearly with repeated co-occurrence. Second, we developed a computational model, SEMANT, suggesting a possible mechanism for this semantic episodic interaction. In SEMANT, episodic associations are implemented through lateral connections between nodes in a pre-existent self-organized map of word semantics. These connections are strengthened at each instance of concomitant activation, proportionally with the amount of the overlapping activity waves of activated nodes. In computer simulations SEMANT replicated the dynamics of associative learning in humans and led to testable predictions concerning normal associative learning as well as impaired learning in a diffuse semantic system like that characteristic of schizophrenia. PMID- 21635312 TI - Understanding the emergence of modularity in neural systems. AB - Modularity in the human brain remains a controversial issue, with disagreement over the nature of the modules that exist, and why, when, and how they emerge. It is a natural assumption that modularity offers some form of computational advantage, and hence evolution by natural selection has translated those advantages into the kind of modular neural structures familiar to cognitive scientists. However, simulations of the evolution of simplified neural systems have shown that, in many cases, it is actually non-modular architectures that are most efficient. In this paper, the relevant issues are discussed and a series of simulations are presented that reveal crucial dependencies on the details of the learning algorithms and tasks that are being modelled, and the importance of taking into account known physical brain constraints, such as the degree of neural connectivity. A pattern is established which provides one explanation of why modularity should emerge reliably across a range of neural processing tasks. PMID- 21635313 TI - Recursion, language, and starlings. AB - It has been claimed that recursion is one of the properties that distinguishes human language from any other form of animal communication. Contrary to this claim, a recent study purports to demonstrate center-embedded recursion in starlings. I show that the performance of the birds in this study can be explained by a counting strategy, without any appreciation of center-embedding. To demonstrate that birds understand center-embedding of sequences of the form A(n) B(n) (such as A(1) A(2) B(2) B(1) , or A(3) A(4) A(5) B(5) B(4) B(3) ) would require not only that they discriminate such patterns from other patterns, but that they appreciate that elements must be bound from the outside in (thus, in the above examples, A(1) B(1) , A(2) B(2) , A(3) B(3) , A(4) B(4) , A(5) B(5) are bound pairs). This has not been shown in nonhuman species, and sentences with this structure are difficult even for humans to parse. There appears to be no evidence to date that nonhuman species understand recursion. PMID- 21635314 TI - Attention to endpoints: a cross-linguistic constraint on spatial meaning. AB - We investigate a possible universal constraint on spatial meaning. It has been proposed that people attend preferentially to the endpoints of spatial motion events, and that languages may therefore make finer semantic distinctions at event endpoints than at event beginnings. We test this proposal. In Experiment 1, we show that people discriminate the endpoints of spatial motion events more readily than they do event beginnings-suggesting a non-linguistic attentional bias toward endpoints. In Experiment 2, speakers of Arabic, Chinese, and English each described a set of spatial events displayed in video clips. Although the spatial systems of these languages differ, speakers of all three languages made finer semantic distinctions at event endpoints, compared to event beginnings. These findings are consistent with the proposal that event endpoints are privileged over event beginnings, in both language and perception. PMID- 21635315 TI - Real and imagined body movement primes metaphor comprehension. AB - We demonstrate in two experiments that real and imagined body movements appropriate to metaphorical phrases facilitate people's immediate comprehension of these phrases. Participants first learned to make different body movements given specific cues. In two reading time studies, people were faster to understand a metaphorical phrase, such as push the argument, when they had previously just made an appropriate body action (e.g., a push movement) (Experiment 1), or imagined making a specific body movement (Experiment 2), than when they first made a mismatching body action (e.g., a chewing movement) or no movement. These findings support the idea that appropriate body action, or even imagined action, enhances people's embodied, metaphorical construal of abstract concepts that are referred to in metaphorical phrases. PMID- 21635316 TI - Spatial and linguistic aspects of visual imagery in sentence comprehension. AB - There is mounting evidence that language comprehension involves the activation of mental imagery of the content of utterances (Barsalou, 1999; Bergen, Chang, & Narayan, 2004; Bergen, Narayan, & Feldman, 2003; Narayan, Bergen, & Weinberg, 2004; Richardson, Spivey, McRae, & Barsalou, 2003; Stanfield & Zwaan, 2001; Zwaan, Stanfield, & Yaxley, 2002). This imagery can have motor or perceptual content. Three main questions about the process remain under-explored, however. First, are lexical associations with perception or motion sufficient to yield mental simulation, or is the integration of lexical semantics into larger structures, like sentences, necessary? Second, what linguistic elements (e.g., verbs, nouns, etc.) trigger mental simulations? Third, how detailed are the visual simulations that are performed? A series of behavioral experiments address these questions, using a visual object categorization task to investigate whether up- or down-related language selectively interferes with visual processing in the same part of the visual field (following Richardson et al., 2003). The results demonstrate that either subject nouns or main verbs can trigger visual imagery, but only when used in literal sentences about real space-metaphorical language does not yield significant effects-which implies that it is the comprehension of the sentence as a whole and not simply lexical associations that yields imagery effects. These studies also show that the evoked imagery contains detail as to the part of the visual field where the described scene would take place. PMID- 21635317 TI - Adaptive non-interventional heuristics for covariation detection in causal induction: model comparison and rational analysis. AB - In this article, 41 models of covariation detection from 2 * 2 contingency tables were evaluated against past data in the literature and against data from new experiments. A new model was also included based on a limiting case of the normative phi-coefficient under an extreme rarity assumption, which has been shown to be an important factor in covariation detection (McKenzie & Mikkelsen, 2007) and data selection (Hattori, 2002; Oaksford & Chater, 1994, 2003). The results were supportive of the new model. To investigate its explanatory adequacy, a rational analysis using two computer simulations was conducted. These simulations revealed the environmental conditions and the memory restrictions under which the new model best approximates the normative model of covariation detection in these tasks. They thus demonstrated the adaptive rationality of the new model. PMID- 21635318 TI - Representation, interaction, and intersubjectivity. AB - What the participants share, their common "sense" of the world, creates a foundation, a framing, an orientation that enables human actors to see and act in coordination with one another. For recurrent activities, the methods the participants use to understand each other as they act change, making the intersubjective space in which actors operate richer and easier to produce. This article works through some of the issues that emerge from a close examination of intersubjectivity as it is managed through representation and interaction. The data that are presented document, in detail, a sequence of related interactions, within and across episodes of cooperation, where continuity and change can be observed. The emergence of conversational structure and coordinating representations are significant milestones in the long-term development of a representational practice that support the runtime co-construction of intersubjective space. Conversational structures emerge interactively to mediate recurrent points of coordination in the domain activity, and only secondarily the conversation itself. Coordinating representations change the representational practice of the participants by making it easier to manage their "shared view" of the collective work, enabling the participants to make progress, expand the field of the common activity, while exhibiting more control of if and when explicit grounding occurs. PMID- 21635319 TI - "What if...": The Use of Conceptual Simulations in Scientific Reasoning. AB - The term conceptual simulation refers to a type of everyday reasoning strategy commonly called "what if" reasoning. It has been suggested in a number of contexts that this type of reasoning plays an important role in scientific discovery; however, little direct evidence exists to support this claim. This article proposes that conceptual simulation is likely to be used in situations of informational uncertainty, and may be used to help scientists resolve that uncertainty. We conducted two studies to investigate the relationship between conceptual simulation and informational uncertainty. Study 1 was an in vivo study of expert scientists; the results suggest that scientists do use conceptual simulation in situations of informational uncertainty, and that they use conceptual simulation to make inferences from their data using the analogical reasoning process of alignment by similarity detection. Study 2 experimentally manipulated experts' level of uncertainty and provides further support for the hypothesis that conceptual simulation is more likely to be used in situations of informational uncertainty. Finally, we discuss the relationship between conceptual simulation and other types of reasoning using qualitative mental models. PMID- 21635320 TI - Mapping the color space of saccadic selectivity in visual search. AB - Color coding is used to guide attention in computer displays for such critical tasks as baggage screening or air traffic control. It has been shown that a display object attracts more attention if its color is more similar to the color for which one is searching. However, what does similar precisely mean? Can we predict the amount of attention that a display color will receive during a search for a given target color? To tackle this question, two color-search experiments measuring the selectivity of saccadic eye movements and mapping out its underlying color space were conducted. A variety of mathematical models, predicting saccadic selectivity for given target and display colors, were devised and evaluated. The results suggest that applying a Gaussian function to a weighted Euclidean distance in a slightly modified HSI color space is the best predictor of saccadic selectivity in the chosen paradigm. Hue and intensity information by itself provides a basis for useful predictors, spanning a possibly spherical color space of saccadic selectivity. Although the current models cannot predict saccadic selectivity values for a wide variety of visual search tasks, they reveal some characteristics of color search that are of both theoretical and applied interest, such as for the design of human-computer interfaces. PMID- 21635321 TI - Tracking the continuity of language comprehension: computer mouse trajectories suggest parallel syntactic processing. AB - Although several theories of online syntactic processing assume the parallel activation of multiple syntactic representations, evidence supporting simultaneous activation has been inconclusive. Here, the continuous and non ballistic properties of computer mouse movements are exploited, by recording their streaming x, y coordinates to procure evidence regarding parallel versus serial processing. Participants heard structurally ambiguous sentences while viewing scenes with properties either supporting or not supporting the difficult modifier interpretation. The curvatures of the elicited trajectories revealed both an effect of visual context and graded competition between simultaneously active syntactic representations. The results are discussed in the context of 3 major groups of theories within the domain of sentence processing. PMID- 21635322 TI - Race categorization modulates holistic face encoding. AB - Recent studies have shown that same-race (SR) faces are processed more holistically than other-race (OR) faces, a difference that may underlie the greater difficulty at recognizing OR than SR faces (the "other-race effect"). This article provides original evidence suggesting that the holistic processing of faces may be sensitive to the observers' racial categorization of the face. In Experiment 1, Caucasian participants performed a face-composite task with Caucasian faces, Asian faces, and racially ambiguous morphed face stimuli. Identical morphed face stimuli were processed more holistically when categorized as SR than as OR faces. Experiment 2 further suggests that this finding was not underlain by strategic or training effects. Overall, these results support the view that one's categorization of a face as belonging to the same or another race plays a critical role in the holistic processing of this face. PMID- 21635323 TI - Linguistic self-correction in the absence of feedback: a new approach to the logical problem of language acquisition. AB - In a series of studies children show increasing mastery of irregular plural forms (such as mice) simply by producing erroneous over-regularized versions of them (such as mouses). We explain this phenomenon in terms of successive approximation in imitation: Children over-regularize early in acquisition because the representations of frequent, regular plural forms develop more quickly, such that at the earliest stages of production they interfere with children's attempts to imitatively reproduce irregular forms they have heard in the input. As the strength of the representations that determine children's productions settle asymptotically, the early advantage for the frequent regular forms is negated, and children's attempts to imitate the irregular forms they have observed become more likely to succeed (a process that produces the classic U-shape in children's acquisition of plural inflection). These data show that children can acquire correct linguistic behavior without feedback in a situation where, as a result of philosophical and linguistic analyses, it has often been argued that it is logically impossible for them to do so. PMID- 21635324 TI - Foundations of representation: where might graphical symbol systems come from? AB - It has been suggested that iconic graphical signs evolve into symbolic graphical signs through repeated usage. This article reports a series of interactive graphical communication experiments using a 'pictionary' task to establish the conditions under which the evolution might occur. Experiment 1 rules out a simple repetition based account in favor of an account that requires feedback and interaction between communicators. Experiment 2 shows how the degree of interaction affects the evolution of signs according to a process of grounding. Experiment 3 confirms the prediction that those not involved directly in the interaction have trouble interpreting the graphical signs produced in Experiment 1. On the basis of these results, this article argues that icons evolve into symbols as a consequence of the systematic shift in the locus of information from the sign to the users' memory of the sign's usage supported by an interactive grounding process. PMID- 21635325 TI - Understanding our understanding of strategic scenarios: what role do chunks play? AB - There is a crucial debate concerning the nature of chess chunks: One current possibility states that chunks are built by encoding particular combinations of pieces-on-squares (POSs), and that chunks are formed mostly by "close" pieces (in a "Euclidean" sense). A complementary hypothesis is that chunks are encoded by abstract, semantic information. This article extends recent experiments and shows that chess players are able to perceive strong similarity between very different positions if the pieces retain the same abstract roles in both of them. This casts doubt on the idea that POS information is the key information encoded in chess chunks, and this article proposes, instead, that the key encoding involves the abstract roles that pieces (and sets of pieces) play-a theoretical standpoint in line with the research program in semantics that places analogy at the core of cognition. PMID- 21635326 TI - Mnemonic context effect in two cultures: attention to memory representations? AB - In two experiments we demonstrate a substantial cross-cultural difference in a mnemonic context effect, whereby a magnitude estimate of a simple stimulus such as a line or circle is biased toward the center of the distribution of previously seen instances of the same class. In support of the hypothesis that Asians are more likely than Americans to disperse their attention to both the target stimulus and its mnemonic context, this effect was consistently larger for Japanese than for Americans. Moreover, the cultural difference was attenuated by an experimentally induced belief in class homogeneity that augmented the context effect itself in both cultures. More important, these belief effects happened in the absence of any objective change in stimulus distribution. Implications for sociocultural shaping of cognition are discussed. PMID- 21635327 TI - Extending the e-z reader model of eye movement control to chinese readers. AB - Chinese readers' eye movements were simulated in the context of the E-Z Reader model, which was developed to account for the eye movements of readers of English. Despite obvious differences between English and Chinese, the model did a fairly good job of simulating the eye movements of Chinese readers. The successful simulation suggests that the control of eye movements in reading Chinese is similar to that in an alphabetic language such as English. PMID- 21635329 TI - Introduction to the 2006 rumelhart prize special issue honoring roger shepard. PMID- 21635328 TI - The brain is both neurocomputer and quantum computer. AB - In their article, Is the Brain a Quantum Computer,? Litt, Eliasmith, Kroon, Weinstein, and Thagard (2006) criticize the Penrose-Hameroff "Orch OR" quantum computational model of consciousness, arguing instead for neurocomputation as an explanation for mental phenomena. Here I clarify and defend Orch OR, show how Orch OR and neurocomputation are compatible, and question whether neurocomputation alone can physiologically account for coherent gamma synchrony EEG, a candidate for the neural correlate of consciousness. Orch OR is based on quantum computation in microtubules within dendrites in cortex and other regions linked by dendritic-dendritic gap junctions ("dendritic webs") acting as laterally connected input layers of the brain's neurocomputational architecture. Within dendritic webs, consciousness is proposed to occur as gamma EEG synchronized sequences of discrete quantum computational events acting in integration phases of neurocomputational "integrate-and-fire" cycles. Orch OR is a viable approach toward understanding how the brain produces consciousness. PMID- 21635330 TI - The step to rationality: the efficacy of thought experiments in science, ethics, and free will. AB - Examples from Archimedes, Galileo, Newton, Einstein, and others suggest that fundamental laws of physics were-or, at least, could have been-discovered by experiments performed not in the physical world but only in the mind. Although problematic for a strict empiricist, the evolutionary emergence in humans of deeply internalized implicit knowledge of abstract principles of transformation and symmetry may have been crucial for humankind's step to rationality-including the discovery of universal principles of mathematics, physics, ethics, and an account of free will that is compatible with determinism. PMID- 21635331 TI - From universal laws of cognition to specific cognitive models. AB - The remarkable successes of the physical sciences have been built on highly general quantitative laws, which serve as the basis for understanding an enormous variety of specific physical systems. How far is it possible to construct universal principles in the cognitive sciences, in terms of which specific aspects of perception, memory, or decision making might be modelled? Following Shepard (e.g., 1987), it is argued that some universal principles may be attainable in cognitive science. Here, 2 examples are proposed: the simplicity principle (which states that the cognitive system prefers patterns that provide simpler explanations of available data); and the scale-invariance principle, which states that many cognitive phenomena are independent of the scale of relevant underlying physical variables, such as time, space, luminance, or sound pressure. This article illustrates how principles may be combined to explain specific cognitive processes by using these principles to derive SIMPLE, a formal model of memory for serial order (Brown, Neath, & Chater, 2007), and briefly mentions some extensions to models of identification and categorization. This article also considers the scope and limitations of universal laws in cognitive science. PMID- 21635332 TI - Using category structures to test iterated learning as a method for identifying inductive biases. AB - Many of the problems studied in cognitive science are inductive problems, requiring people to evaluate hypotheses in the light of data. The key to solving these problems successfully is having the right inductive biases-assumptions about the world that make it possible to choose between hypotheses that are equally consistent with the observed data. This article explores a novel experimental method for identifying the biases that guide human inductive inferences. The idea behind this method is simple: This article uses the responses produced by a participant on one trial to generate the stimuli that either they or another participant will see on the next. A formal analysis of this "iterated learning" procedure, based on the assumption that the learners are Bayesian agents, predicts that it should reveal the inductive biases of these learners, as expressed in a prior probability distribution over hypotheses. This article presents a series of experiments using stimuli based on a well-studied set of category structures, demonstrating that iterated learning can be used to reveal the inductive biases of human learners. PMID- 21635333 TI - A rational analysis of rule-based concept learning. AB - This article proposes a new model of human concept learning that provides a rational analysis of learning feature-based concepts. This model is built upon Bayesian inference for a grammatically structured hypothesis space-a concept language of logical rules. This article compares the model predictions to human generalization judgments in several well-known category learning experiments, and finds good agreement for both average and individual participant generalizations. This article further investigates judgments for a broad set of 7-feature concepts a more natural setting in several ways-and again finds that the model explains human performance. PMID- 21635334 TI - Is a single-bladed knife enough to dissect human cognition? Commentary on griffiths et Al. AB - Griffiths, Christian, and Kalish (this issue) present an iterative-learning paradigm applying a Bayesian model to understand inductive biases in categorization. The authors argue that the paradigm is useful as an exploratory tool to understand inductive biases in situations where little is known about the task. It is argued that a theory developed only at the computational level is much like a single-bladed knife that is only useful in highly idealized situations. To be useful as a general tool that cuts through the complex fabric of cognition, we need at least two-bladed scissors that combine both computational and psychological constraints to characterize human behavior. To temper its sometimes expansive claims, it is time to show what a Bayesian model cannot explain. Insight as to how human reality may differ from the Bayesian predictions may shed more light on human cognition than the simpler focus on what the Bayesian approach can explain. There remains much to be done in terms of integrating Bayesian approaches and other approaches in modeling human cognition. PMID- 21635335 TI - Observation can be as effective as action in problem solving. AB - This study discusses findings that replicate and extend the original work of Burns and Vollmeyer (2002), which showed that performance in problem-solving tasks was more accurate when people were engaged in a non-specific goal than in a specific goal. The main innovation here was to examine the goal specificity effect under both observation-based and conventional action-based learning conditions. The findings show that goal specificity affects the accuracy of problem solving in the same way when the learning stage of the task is observation-based as when it is action-based. In addition, the findings show that, when instructions do not promote goal specificity, observation-based problem solving is as effective as action-based problem solving. PMID- 21635336 TI - Lexical categories at the edge of the word. AB - Language acquisition may be one of the most difficult tasks that children face during development. They have to segment words from fluent speech, figure out the meanings of these words, and discover the syntactic constraints for joining them together into meaningful sentences. Over the past couple of decades, computational modeling has emerged as a new paradigm for gaining insights into the mechanisms by which children may accomplish these feats. Unfortunately, many of these models assume a computational complexity and linguistic knowledge likely to be beyond the abilities of developing young children. This article shows that, using simple statistical procedures, significant correlations exist between the beginnings and endings of a word and its lexical category in English, Dutch, French, and Japanese. Therefore, phonetic information can contribute to individuating higher level structural properties of these languages. This article also presents a simple 2-layer connectionist model that, once trained with an initial small sample of words labeled for lexical category, can infer the lexical category of a large proportion of novel words using only word-edge phonological information, namely the first and last phoneme of a word. The results suggest that simple procedures combined with phonetic information perceptually available to children provide solid scaffolding for emerging lexical categories in language development. PMID- 21635337 TI - Is Structure Dependence an Innate Constraint? New Experimental Evidence From Children's Complex-Question Production. AB - According to Crain and Nakayama (1987), when forming complex yes/no questions, children do not make errors such as Is the boy who smoking is crazy? because they have innate knowledge of structure dependence and so will not move the auxiliary from the relative clause. However, simple recurrent networks are also able to avoid such errors, on the basis of surface distributional properties of the input (Lewis & Elman, 2001; Reali & Christiansen, 2005). Two new elicited production studies revealed that (a) children occasionally produce structure-dependence errors and (b) the pattern of children's auxiliary-doubling errors (Is the boy who is smoking is crazy?) suggests a sensitivity to surface co-occurrence patterns in the input. This article concludes that current data do not provide any support for the claim that structure dependence is an innate constraint, and that it is possible that children form a structure-dependent grammar on the basis of exposure to input that exhibits this property. PMID- 21635338 TI - Observing tutorial dialogues collaboratively: insights about human tutoring effectiveness from vicarious learning. AB - The goals of this study are to evaluate a relatively novel learning environment, as well as to seek greater understanding of why human tutoring is so effective. This alternative learning environment consists of pairs of students collaboratively observing a videotape of another student being tutored. Comparing this collaboratively observing environment to four other instructional methods one-on-one human tutoring, observing tutoring individually, collaborating without observing, and studying alone-the results showed that students learned to solve physics problems just as effectively from observing tutoring collaboratively as the tutees who were being tutored individually. We explain the effectiveness of this learning environment by postulating that such a situation encourages learners to become active and constructive observers through interactions with a peer. In essence, collaboratively observing combines the benefit of tutoring with the benefit of collaborating. The learning outcomes of the tutees and the collaborative observers, along with the tutoring dialogues, were used to further evaluate three hypotheses explaining why human tutoring is an effective learning method. Detailed analyses of the protocols at several grain sizes suggest that tutoring is effective when tutees are independently or jointly constructing knowledge: with the tutor, but not when the tutor independently conveys knowledge. PMID- 21635339 TI - The role of words and sounds in infants' visual processing: from overshadowing to attentional tuning. AB - Although it is well documented that language plays an important role in cognitive development, there are different views concerning the mechanisms underlying these effects. Some argue that even early in development, effects of words stem from top-down knowledge, whereas others argue that these effects stem from auditory input affecting attention allocated to visual input. Previous research (e.g., Robinson & Sloutsky, 2004a) demonstrated that non-speech sounds attenuate processing of corresponding visual input at 8, 12, and 16 months of age, whereas the current study demonstrates that words attenuate visual processing at 10 months but not at 16 months (Experiment 1). Furthermore, prefamiliarization with non-speech sounds (Experiment 2) resulted in able processing of visual input by 16-month-olds. These findings suggest that some effects of labels found early in development may stem from familiarity with human speech. The possibility of general-auditory factors underlying the effects of words on cognitive development is discussed. PMID- 21635340 TI - Trade-offs between grounded and abstract representations: evidence from algebra problem solving. AB - This article explores the complementary strengths and weaknesses of grounded and abstract representations in the domain of early algebra. Abstract representations, such as algebraic symbols, are concise and easy to manipulate but are distanced from any physical referents. Grounded representations, such as verbal descriptions of situations, are more concrete and familiar, and they are more similar to physical objects and everyday experience. The complementary computational characteristics of grounded and abstract representations lead to trade-offs in problem-solving performance. In prior research with high school students solving relatively simple problems, Koedinger and Nathan (2004) demonstrated performance benefits of grounded representations over abstract representations-students were better at solving simple story problems than the analogous equations. This article extends this prior work to examine both simple and more complex problems in two samples of college students. On complex problems with two references to the unknown, a "symbolic advantage" emerged, such that students were better at solving equations than analogous story problems. Furthermore, the previously observed "verbal advantage" on simple problems was replicated. We thus provide empirical support for a trade-off between grounded, verbal representations, which show advantages on simpler problems, and abstract, symbolic representations, which show advantages on more complex problems. PMID- 21635341 TI - A probabilistic model of melody perception. AB - This study presents a probabilistic model of melody perception, which infers the key of a melody and also judges the probability of the melody itself. The model uses Bayesian reasoning: For any "surface" pattern and underlying "structure," we can infer the structure maximizing P(structure|surface) based on knowledge of P(surface, structure). The probability of the surface can then be calculated as ? P(surface, structure), summed over all structures. In this case, the surface is a pattern of notes; the structure is a key. A generative model is proposed, based on three principles: (a) melodies tend to remain within a narrow pitch range; (b) note-to-note intervals within a melody tend to be small; and (c) notes tend to conform to a distribution (or key profile) that depends on the key. The model is tested in three ways. First, it is tested on its ability to identify the keys of a set of folksong melodies. Second, it is tested on a melodic expectation task in which it must judge the probability of different notes occurring given a prior context; these judgments are compared with perception data from a melodic expectation experiment. Finally, the model is tested on its ability to detect incorrect notes in melodies by assigning them lower probabilities than the original versions. PMID- 21635342 TI - Understanding and using principles of arithmetic: operations involving negative numbers. AB - Previous work has investigated adults' knowledge of principles for arithmetic with positive numbers (Dixon, Deets, & Bangert, 2001). The current study extends this past work to address adults' knowledge of principles of arithmetic with a negative number, and also investigates links between knowledge of principles and problem representation. Participants (N = 44) completed two tasks. In the Evaluation task, participants rated how well sets of equations were solved. Some sets violated principles of arithmetic and others did not. Participants rated non violation sets higher than violation sets for two different principles for subtraction with a negative number. In the Word Problem task, participants read word problems and set up equations that could be used to solve them. Participants who displayed greater knowledge of principles of arithmetic with a negative number were more likely to set up equations that involved negative numbers. Thus, participants' knowledge of arithmetic principles was related to their problem representations. PMID- 21635343 TI - Perceiving the present and a systematization of illusions. AB - Over the history of the study of visual perception there has been great success at discovering countless visual illusions. There has been less success in organizing the overwhelming variety of illusions into empirical generalizations (much less explaining them all via a unifying theory). Here, this article shows that it is possible to systematically organize more than 50 kinds of illusion into a 7 * 4 matrix of 28 classes. In particular, this article demonstrates that (1) smaller sizes, (2) slower speeds, (3) greater luminance contrast, (4) farther distance, (5) lower eccentricity, (6) greater proximity to the vanishing point, and (7) greater proximity to the focus of expansion all tend to have similar perceptual effects, namely, to (A) increase perceived size, (B) increase perceived speed, (C) decrease perceived luminance contrast, and (D) decrease perceived distance. The detection of these empirical regularities was motivated by a hypothesis, called "perceiving the present," that the visual system possesses mechanisms for compensating neural delay during forward motion. This article shows how this hypothesis predicts the empirical regularity. PMID- 21635344 TI - Bootstrapping cognition from behavior-a computerized thought experiment. AB - We show that simple perceptual competences can emerge from an internal simulation of action effects and are thus grounded in behavior. A simulated agent learns to distinguish between dead ends and corridors without the necessity to represent these concepts in the sensory domain. Initially, the agent is only endowed with a simple value system and the means to extract low-level features from an image. In the interaction with the environment, it acquires a visuo-tactile forward model that allows the agent to predict how the visual input is changing under its movements, and whether movements will lead to a collision. From short-term predictions based on the forward model, the agent learns an inverse model. The inverse model in turn produces suggestions about which actions should be simulated in long-term predictions, and long-term predictions eventually give rise to the perceptual ability. PMID- 21635345 TI - The weckud wetch of the wast: lexical adaptation to a novel accent. AB - Two experiments investigated the mechanism by which listeners adjust their interpretation of accented speech that is similar to a regional dialect of American English. Only a subset of the vowels of English (the front vowels) were shifted during adaptation, which consisted of listening to a 20-min segment of the "Wizard of Oz." Compared to a baseline (unadapted) condition, listeners showed significant adaptation to the accented speech, as indexed by increased word judgments on a lexical decision task. Adaptation also generalized to test words that had not been presented in the accented passage but that contained the shifted vowels. A control experiment showed that the adaptation effect was specific to the direction of the shift in the vowel space and not to a general relaxation of the criterion for what constitutes a good exemplar of the accented vowel category. Taken together, these results provide evidence for a context specific vowel adaptation mechanism that enables a listener to adjust to the dialect of a particular talker. PMID- 21635346 TI - Congruity Effects in Time and Space: Behavioral and ERP Measures. AB - Two experiments investigated whether motion metaphors for time affected the perception of spatial motion. Participants read sentences either about literal motion through space or metaphorical motion through time written from either the ego-moving or object-moving perspective. Each sentence was followed by a cartoon clip. Smiley-moving clips showed an iconic happy face moving toward a polygon, and shape-moving clips showed a polygon moving toward a happy face. In Experiment 1, using an explicit judgment task, participants judged smiley-moving cartoons as related to ego-moving sentences about space and about time, and shape-moving cartoons as related to object-moving sentences. In Experiment 2, participants viewed the same stimuli, but the cartoons were task-irrelevant. Event-related brain potentials revealed an early attentional effect of congruity on cartoons following sentences about space, and a later semantic effect on cartoons following sentences about time. Results are most consistent with accounts that posit differences in the processing of novel and conventional metaphors. PMID- 21635347 TI - Perceptual processing affects conceptual processing. AB - According to the Perceptual Symbols Theory of cognition (Barsalou, 1999), modality-specific simulations underlie the representation of concepts. A strong prediction of this view is that perceptual processing affects conceptual processing. In this study, participants performed a perceptual detection task and a conceptual property-verification task in alternation. Responses on the property verification task were slower for those trials that were preceded by a perceptual trial in a different modality than for those that were preceded by a perceptual trial in the same modality. This finding of a modality-switch effect across perceptual processing and conceptual processing supports the hypothesis that perceptual and conceptual representations are partially based on the same systems. PMID- 21635348 TI - The benefits of sensorimotor knowledge: body-object interaction facilitates semantic processing. AB - This article examined the effects of body-object interaction (BOI) on semantic processing. BOI measures perceptions of the ease with which a human body can physically interact with a word's referent. In Experiment 1, BOI effects were examined in 2 semantic categorization tasks (SCT) in which participants decided if words are easily imageable. Responses were faster and more accurate for high BOI words (e.g., mask) than for low BOI words (e.g., ship). In Experiment 2, BOI effects were examined in a semantic lexical decision task (SLDT), which taps both semantic feedback and semantic processing. The BOI effect was larger in the SLDT than in the SCT, suggesting that BOI facilitates both semantic feedback and semantic processing. The findings are consistent with the embodied cognition perspective (e.g., Barsalou's, 1999, Perceptual Symbols Theory), which proposes that sensorimotor interactions with the environment are incorporated in semantic knowledge. PMID- 21635349 TI - Bewitchment, biology, or both: the co-existence of natural and supernatural explanatory frameworks across development. AB - Three studies examined the co-existence of natural and supernatural explanations for illness and disease transmission, from a developmental perspective. The participants (5-, 7-, 11-, and 15-year-olds and adults; N = 366) were drawn from 2 Sesotho-speaking South African communities, where Western biomedical and traditional healing frameworks were both available. Results indicated that, although biological explanations for illness were endorsed at high levels, witchcraft was also often endorsed. More important, bewitchment explanations were neither the result of ignorance nor replaced by biological explanations. Instead, both natural and supernatural explanations were used to explain the same phenomena, and bewitchment explanations were highest among adults. Taken together, these data provide insight into how diverse, culturally constructed belief systems about illness co-exist across development. PMID- 21635350 TI - Processing polarity: how the ungrammatical intrudes on the grammatical. AB - A central question in online human sentence comprehension is, "How are linguistic relations established between different parts of a sentence?" Previous work has shown that this dependency resolution process can be computationally expensive, but the underlying reasons for this are still unclear. This article argues that dependency resolution is mediated by cue-based retrieval, constrained by independently motivated working memory principles defined in a cognitive architecture. To demonstrate this, this article investigates an unusual instance of dependency resolution, the processing of negative and positive polarity items, and confirms a surprising prediction of the cue-based retrieval model: Partial cue matches-which constitute a kind of similarity-based interference-can give rise to the intrusion of ungrammatical retrieval candidates, leading to both processing slow-downs and even errors of judgment that take the form of illusions of grammaticality in patently ungrammatical structures. A notable achievement is that good quantitative fits are achieved without adjusting the key model parameters. PMID- 21635351 TI - Understanding mortality and the life of the ancestors in rural madagascar. AB - Across two studies, a wide age range of participants was interviewed about the nature of death. All participants were living in rural Madagascar in a community where ancestral beliefs and practices are widespread. In Study 1, children (8-17 years) and adults (19-71 years) were asked whether bodily and mental processes continue after death. The death in question was presented in the context of a narrative that focused either on the corpse or on the ancestral practices associated with the afterlife. Participants aged 8 years and older claimed that death brings an end to most bodily and mental processes. Nevertheless, particularly in the context of the religious narrative, they claimed that certain mental processes continue even after death. This assertion of an afterlife was more evident among adults than children, especially with respect to cognitive processes, such as knowing and remembering. In Study 2, 5- and 7-year-olds were asked similar questions in connection with the death of a bird and a person. Seven-year-olds consistently claimed that bodily and mental processes cease at death, whereas 5-year-olds were unsystematic in their replies. Together, the two studies replicate and extend findings obtained with Western children showing that, in the course of development, different conceptions of death are elaborated a biological conception in which death terminates living processes and a religious conception in which death marks the beginning of a new form of spiritual existence. PMID- 21635352 TI - Working memory in wayfinding-a dual task experiment in a virtual city. AB - This study examines the working memory systems involved in human wayfinding. In the learning phase, 24 participants learned two routes in a novel photorealistic virtual environment displayed on a 220 degrees screen while they were disrupted by a visual, a spatial, a verbal, or-in a control group-no secondary task. In the following wayfinding phase, the participants had to find and to "virtually walk" the two routes again. During this wayfinding phase, a number of dependent measures were recorded. This research shows that encoding wayfinding knowledge interfered with the verbal and with the spatial secondary task. These interferences were even stronger than the interference of wayfinding knowledge with the visual secondary task. These findings are consistent with a dual-coding approach of wayfinding knowledge. PMID- 21635353 TI - Bigrams and the richness of the stimulus. AB - Recent challenges to Chomsky's poverty of the stimulus thesis for language acquisition suggest that children's primary data may carry "indirect evidence" about linguistic constructions despite containing no instances of them. Indirect evidence is claimed to suffice for grammar acquisition, without need for innate knowledge. This article reports experiments based on those of Reali and Christiansen (2005), who demonstrated that a simple bigram language model can induce the correct form of auxiliary inversion in certain complex questions. This article investigates the nature of the indirect evidence that supports this learning, and assesses how reliably it is available. Results confirm the original finding for one specific sentence type but show that the model's success is highly circumscribed. It performs poorly on inversion in related constructions in English and Dutch. Because other, more powerful statistical models have so far been shown to succeed only on the same limited subset of cases as the bigram model, it remains to be seen whether stimulus richness can be substantiated more generally. PMID- 21635354 TI - A computational model of early argument structure acquisition. AB - How children go about learning the general regularities that govern language, as well as keeping track of the exceptions to them, remains one of the challenging open questions in the cognitive science of language. Computational modeling is an important methodology in research aimed at addressing this issue. We must determine appropriate learning mechanisms that can grasp generalizations from examples of specific usages, and that exhibit patterns of behavior over the course of learning similar to those in children. Early learning of verb argument structure is an area of language acquisition that provides an interesting testbed for such approaches due to the complexity of verb usages. A range of linguistic factors interact in determining the felicitous use of a verb in various constructions-associations between syntactic forms and properties of meaning that form the basis for a number of linguistic and psycholinguistic theories of language. This article presents a computational model for the representation, acquisition, and use of verbs and constructions. The Bayesian framework is founded on a novel view of constructions as a probabilistic association between syntactic and semantic features. The computational experiments reported here demonstrate the feasibility of learning general constructions, and their exceptions, from individual usages of verbs. The behavior of the model over the timecourse of acquisition mimics, in relevant aspects, the stages of learning exhibited by children. Therefore, this proposal sheds light on the possible mechanisms at work in forming linguistic generalizations and maintaining knowledge of exceptions. PMID- 21635355 TI - Strategy generalization across orientation tasks: testing a computational cognitive model. AB - Humans use their spatial information processing abilities flexibly to facilitate problem solving and decision making in a variety of tasks. This article explores the question of whether a general strategy can be adapted for performing two different spatial orientation tasks by testing the predictions of a computational cognitive model. Human performance was measured on an orientation task requiring participants to identify the location of a target either on a map (find-on-map) or within an egocentric view of a space (find-in-scene). A general strategy instantiated in a computational cognitive model of the find-on-map task, based on the results from Gunzelmann and Anderson (2006), was adapted to perform both tasks and used to generate performance predictions for a new study. The qualitative fit of the model to the human data supports the view that participants were able to tailor a general strategy to the requirements of particular spatial tasks. The quantitative differences between the predictions of the model and the performance of human participants in the new experiment expose individual differences in sample populations. The model provides a means of accounting for those differences and a framework for understanding how human spatial abilities are applied to naturalistic spatial tasks that involve reasoning with maps. PMID- 21635356 TI - Modeling how, when, and what is learned in a simple fault-finding task. AB - We have developed a process model that learns in multiple ways while finding faults in a simple control panel device. The model predicts human participants' learning through its own learning. The model's performance was systematically compared to human learning data, including the time course and specific sequence of learned behaviors. These comparisons show that the model accounts very well for measures such as problem-solving strategy, the relative difficulty of faults, and average fault-finding time. More important, because the model learns and transfers its learning across problems, it also accounts for the faster problem solving times due to learning when examined across participants, across faults, and across the series of 20 trials on an individual participant basis. The model shows how learning while problem solving can lead to more recognition-based performance, and helps explain how the shape of the learning curve can arise through learning and be modified by differential transfer. Overall, the quality of the correspondence appears to have arisen from procedural, declarative, and episodic learning all taking place within individual problem-solving episodes. PMID- 21635357 TI - The deep versus the shallow: effects of co-speech gestures in learning from discourse. AB - This study concerned the role of gestures that accompany discourse in deep learning processes. We assumed that co-speech gestures favor the construction of a complete mental representation of the discourse content, and we tested the predictions that a discourse accompanied by gestures, as compared with a discourse not accompanied by gestures, should result in better recollection of conceptual information, a greater number of discourse-based inferences drawn from the information explicitly stated in the discourse, and poorer recognition of verbatim of the discourse. The results of three experiments confirmed these predictions. PMID- 21635358 TI - A case-control study of melatonin receptor type 1A polymorphism and acute myocardial infarction in a Spanish population. AB - Coronary artery disease (CAD) is a complex disease with genetic and environmental determinants. Although a large number of genetic polymorphisms involved in the pathogenesis of atherosclerosis have been identified, there is still no evidence of a genetic association with CAD. As melatonin might play a role in the pathogenesis of atherosclerosis through its anti-inflammatory and antioxidant properties, we tested whether the expression of six single nucleotide polymorphisms (SNPs) of the melatonin receptor differs in acute myocardial infarction (AMI) patients with acute myocardial infarction (n = 300) compared with healthy age- and sex-matched controls (n = 250). Finally, only MEL1A receptor SNP rs28383653 was selected because of Hardy-Weinberg equilibrium (chi(2) = 0.49). The distribution of genotype frequencies for this SNP showed that the unfavourable CT genotype was significantly more frequent in patients with AMI than in controls (4.5% versus 1.3%; P = 0.006). Multivariable analysis showed a significantly higher frequency of the unfavourable CT genotype in AMI patients with peripheral arteriopathy (28% versus 10%; P = 0.01). This finding suggests a synergism effect between the unfavourable genotype (CT) of the MELIA receptor SNP and the vascular disease in this subgroup of patients. To our knowledge, this is the first study to report an association between a genetic polymorphism of the melatonin receptor 1A and CAD. PMID- 21635359 TI - St John's wort greatly decreases the plasma concentrations of oral S-ketamine. AB - Ketamine is an intravenous anaesthetic and analgesic agent but it can also be used orally as an adjuvant in the treatment of chronic pain. This study investigated the effect of the herbal antidepressant St John's wort, an inducer of cytochrome P450 3A4 (CYP3A4), on the pharmacokinetics and pharmacodynamics of oral S-ketamine. In a randomized cross-over study with two phases, 12 healthy subjects were pretreated with oral St John's wort or placebo for 14 days. On day 14, they were given an oral dose of 0.3 mg/kg of S-ketamine. Plasma concentrations of ketamine and norketamine were measured for 24 h and pharmacodynamic variables for 12 h. St John's wort decreased the mean area under the plasma concentration-time curve (AUC(0-infinity)) of ketamine by 58% (P < 0.001) and decreased the peak plasma concentration (C(max)) of ketamine by 66% (P < 0.001) when compared with placebo. Mean C(max) of norketamine (the major metabolite of ketamine) was decreased by 23% (P = 0.002) and mean AUC(0-infinity) of norketamine by 18% (P < 0.001) by St John's wort. There was a statistically significant linear correlation between the self-reported drug effect and C(max) of ketamine (r = 0.55; P < 0.01). St John's wort greatly decreased the exposure to oral S-ketamine in healthy volunteers. Although this decrease was not associated with significant changes in the analgesic or behavioural effects of ketamine in the present study, usual doses of S-ketamine may become ineffective if used concomitantly with St John's wort. PMID- 21635360 TI - Assessment of self-administered epinephrine during a training session. PMID- 21635362 TI - A Japanese boy with adenine phosphoribosyltransferase (APRT) deficiency caused by compound heterozygosity including a novel missense mutation in APRT gene. AB - We describe a 2-year-old Japanese boy with radiolucent urolithiasis and recurrent urinary tract infection. Urinalysis showed typical 2,8-dihydroxyadenine (2,8-DHA) crystals, leading to a diagnosis as adenine phosphoribosyltransferase (APRT) deficiency. The sensitivity of proliferating T cells to an adenine analogue, whose cytotoxicity is dependent on APRT, showed that he was homozygous or compound heterozygous for the APRT gene mutation. A genetic analysis revealed a compound heterozygous state for M136T and a novel missense mutation L33P, not previously reported in patients with APRT deficiency. CONCLUSION: Adenine phosphoribosyltransferase deficiency should be suspected in all patients with radiolucent kidney stones, urinary 2,8-DHA crystals were an important finding for an early diagnosis of APRT deficiency. Appropriate treatment should be initiated to prevent the development of urolithiasis or renal failure in APRT-deficient children. The T cell method was useful to detect a homozygote or a compound heterozygote of the pathogenic allelic gene in APRT deficiency, and a genetic analysis revealed a novel mutation L33P. PMID- 21635363 TI - Long-term effects of earlier initiated continuous Kangaroo Mother Care (KMC) for low-birth-weight (LBW) infants in Madagascar. AB - AIM: To examine the long-term effects of earlier initiated continuous Kangaroo Mother Care (KMC) for relatively stable low-birth-weight (LBW) infants in a resource-limited country. METHODS: A randomized controlled trial with long-term follow-up was performed in LBW infants in Madagascar. Earlier continuous KMC (intervention group) was initiated as soon as possible within 24 h postbirth, and later continuous KMC (control group: conventional care) was initiated after complete stabilization. Outcome measures were mortality or readmission, nutritional indicators at 6-12 months postbirth and feeding condition at 6 months postbirth (ClinicalTrials.gov, NCT00531492). RESULTS: A total of 72 infants were followed for mortality or readmission at 6-12 months postbirth. There was no difference between the two groups (7/36 vs. 7/36, Risk ratio (RR), 1.00; 95% CIs, 0.39-2.56; p = 1.00). The proportion of exclusive breast feeding (EBF) at 6 months postbirth was significantly higher with earlier KMC than later KMC (12/29 vs. 4/26; RR 2.69; 95% CIs, 1.00-7.31; p = 0.04). There were no differences in nutritional indicators between the two groups at 6-12 months postbirth. CONCLUSION: Earlier initiated continuous KMC results in a significantly higher proportion of EBF at 6 months postbirth. Further larger-scale long-term evaluations of earlier initiated continuous KMC for LBW infants are needed. PMID- 21635364 TI - Ethical reflections on organ donation from children. PMID- 21635547 TI - Detection of low-level EGFR T790M mutation in lung cancer tissues. AB - Epidermal growth factor receptor (EGFR) gene mutation status is critical to predicting responsiveness to EGFR tyrosine kinase inhibitor (TKI) therapies in non-small cell lung cancer (NSCLC) patients. However, a vast majority of the patients experience recurrence of the cancers by a secondary mutation of EGFR (T790M). Earlier studies suggested evidence that subclones bearing EGFR T790M mutation pre-exist in NSCLCs even prior to the therapies. However, to date, the status of T790M mutation in primary NSCLC is largely known. In this study, we developed an assay using peptide nucleic acid (PNA)-clamping PCR for detection of low-level EGFR T790M mutation. We found that the assay showed the highest sensitivity (0.01% mutation detection) in the clamping condition. We analyzed 147 NSCLC tissues [70 adenocarcinomas (AD), 62 squamous cell carcinomas (SQ), 12 large cell carcinomas (LC), and three adenosquamous carcinomas] that had not been exposed to the TKI therapies, and found 12 (8.2%; 12/147) EGFR T790M mutation in eight AD (11.4%), three SQ (4.8%), and one LC (8.3%) by the PNA-clamping PCR. However, this mutation was not detected by conventional DNA sequencing. Our data indicate that EGFR T790M exists in pretreatment NSCLC at low levels irrespective of histologic types. This study provides a basis for developing an applicable protocol for detecting low-level EGFR T790M mutation in primary NSCLC, which might contribute to predicting recurrence of the tumor in response to the TKI therapies. PMID- 21635548 TI - Microvesicular fat, inter cellular adhesion molecule-1 and regulatory T lymphocytes are of importance for the inflammatory process in livers with non alcoholic steatohepatitis. AB - Great progress has been made in understanding the development of non-alcoholic fatty liver disease (NAFLD) but less is known about the mechanisms underlying the progress from steatosis to steatohepatitis (NASH). Our aim was to evaluate if the amount and type of storage of fat in hepatocytes is of importance for hepatocyte injury. We also wanted to show if not only the innate immunity but also the adaptive immunity is involved in NASH. Thirty-one patients with NASH or borderline NASH and 18 non-NASH patients were investigated. Liver biopsies were scored for NASH according to Kleiner et al. Paraffin-embedded liver biopsies were stained with antibodies against CD3, TLR4, CD68, Cleaved Caspase-3, ICAM1, Foxp3 and ApopTag by immunohistochemistry. Serum soluble ICAM-1 (sICAM-1) were analysed by ELISA. The volume density of fat was 59% in the NASH patients and microvesicular fat, increased in high NAS score patients. ICAM-1 positive hepatocytes were seen in NASH patients and were localized in areas with microvesicular fat. Non-NASH biopsies were negative for ICAM-1 positive hepatocytes. The sICAM-1 were significantly higher in NASH-patients (339.8 +/- 34.07) than in non-NASH patients (229.5 +/- 12.14), p = 0.0015. Patients with NAS score over four had higher area of CD68 positive cells p = 0.0011 and Foxp3 positive cells (p = 0.024) than non-NASH patients. In liver tissue with NASH, hepatocytes with microvesicular steatosis seem to be expressing more inflammatory markers, and in this liver tissue an increased number of CD68 cells and regulatory T-cells (Tregs, e.g. Foxp3+ cells) were seen, indicating an involvement of, both the innate and the adaptive immunity. PMID- 21635549 TI - Mutational analysis of VACM-1/cul5 exons in cancer cell lines. AB - VACM-1, a cul-5 gene product, functions via an E3 ligase complex and when overexpressed, has an antiproliferative effect in many cell types. Overexpression of VACM-/cul5 cDNA mutated at the PKA-specific phosphorylation site at Ser730 reversed this phenotype. These effects are associated with the appearance of larger M(r) species subsequently identified as a Nedd8-modified VACM-1/cul5. Although decreased levels of VACM-1 mRNA detected in several cancers and cancer cell lines may explain the progression of cell growth, possible genetic and epigenetic changes in its sequence have not been analyzed. We hypothesized that in rapidly proliferating cells, VACM-1/cul5 may be mutated at either the PKA specific phosphorylation site or the consensus neddylation site. We used RT-PCR and PCR, to amplify and to sequence mRNA and genomic DNA, respectively. To date we have sequenced all 19 coding exons of the VACM-1/cul5 gene in T47D breast cancer cells, U138MG glioma cells, ACHN renal cancer cells, and OVCAR-3 ovarian cancer cells. Our results indicate that in those cells VACM-1/cul5 is not mutated at the putative phosphorylation or the neddylation site. We have found one silent mutation in the genomic DNA isolated from U138MG, ACHN, and OVCAR-3 cell lines, but not from T47D cells. Our work suggests that in T47D breast cancer cells biologic activity of VACM-1/cul5 may be regulated by posttranslational modifications. PMID- 21635550 TI - Effect of N-nitrosodimethylamine on inducible nitric oxide synthase expression and production of nitric oxide by neutrophils and mononuclear cells: the role of JNK signalling pathway. AB - In neutrophils (PMN) and mononuclear cells (PBMC), one of the enzymes responsible for nitric oxide (NO) synthesis is inducible nitric oxide synthase (iNOS). Changes in iNOS expression result from various signalling pathways including the mitogen-activated protein kinase (MAPK) pathway activated by endogenic and exogenic factors. N-nitrosodimethylamine (NDMA) is a xenobiotic with widespread occurrence in human environment and has an effect on cells of the immune system. The aim of this study was to determine the effect of NDMA on iNOS expression and NO production by human PMN and PBMC cells in the light of superoxide anion production by PMN cells. Moreover, the role of JNK and p38 pathways in NO production with involvement of iNOS was studied. Additionally, the function of JNK pathway in generation of superoxide anion was determined. Moreover, nitrotyrosine expression was studied in PMN and PBMC cells in the presence of NDMA. This work shows that NDMA increases iNOS expression and NO production by PMN and PBMC cells. In addition, elevated expression of phospho-JNK and phospho p38, which are markers of JNK and p38 MAPK pathways activation, were observed. Lower iNOS expression and NO production by neutrophils exposed to extended action of NDMA were observed after application of inhibitor of JNK and p38 pathways. Lower phospho-p38 expression in PMN stimulated by NDMA was found as a result of arresting JNK pathway, whereas, application of inhibitor of p38 pathway resulted in enhanced phospho-JNK expression in PMN and PBMC cells. Increased ability to release superoxide anion by NDMA-stimulated PMN cells was observed. This ability was reduced after the application of inhibitor of JNK pathway. In PMN and PBMC cells exposed to NDMA, an increased expression of nitrotyrosine, which is dependant on JNK and p38 pathways that are activated by this particular xenobiotic, was observed. Generally, increased induction of iNOS related to elevated production of NO by PMN and PBMC cells exposed to NDMA may result in dysfunction of regulation of immunity responses controlled by this molecule in various conditions. Increased expression of nitrotyrosine in PMN and PBMC cells exposed to NDMA may affect their functions in an auto- and/or a paracrine way. Mutual interactions of JNK and p38 MAPK during the induction of iNOS expression in cells exposed to NDMA indicate complex mechanism of induction of iNOS synthase. PMID- 21635551 TI - Pilot study of omega-3 fatty acid supplements in sickle cell disease. AB - In a previous retrospective study, it was observed that the greater the amounts of the omega-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in the blood, the lesser the number of complications of sickle cell disease (SCD) and the higher the steady state haemoglobin level. SCD causes ischaemia reperfusion injury and inflammation; which can be ameliorated by a metabolite of DHA that down-regulates expression of pro-inflammatory genes. The objectives of this prospective pilot study were to evaluate the effects of DHA and EPA supplements in SCD, and test the hypothesis that these effects are mediated partly by reducing inflammation. Oral DHA and EPA supplements were given to 16 SCD patients for 6 months. We then compared pre- and post-supplementation values of number of crisis, steady state Hb, plasma unconjugated bilirubin and three indices of inflammation: plasma interleukin-6, blood neutrophil and platelet counts. There was a significant reduction in the plasma level of unconjugated bilirubin, and the number of sickle cell crisis; but not in the markers of inflammation. The pilot data suggest that DHA and EPA supplements reduce the number of crisis and steady state haemolysis in SCD; but provide no evidence that these effects are mediated by reducing inflammation. PMID- 21635552 TI - Prognostic significance of epidermal growth factor receptor and vascular endothelial growth factor receptor in colorectal adenocarcinoma. AB - The purpose of this study was to evaluate the association between the expression of growth factors and the clinicopathological variables of colorectal adenocarcinoma. Immunohistochemistry and fluorescence in situ hybridization (FISH) were used to evaluate the amplification and expression of epidermal growth factor receptor (EGFR), vascular endothelial growth factor (VEGF), VEGF-D, VEGF receptor (VEGFR)-2, VEGFR-3, transforming growth factor (TGF)-beta1, and insulin like growth factor-1 receptor (IGF-1R) in a tissue microarray of 292 colorectal adenocarcinomas. The expression of EGFR, VEGF, VEGF-D, VEGFR-2 and VEGFR-3 was detected in 5.1%, 10.0%, 6.8%, 5.2%, and 57.2%. EGFR expression was associated with angioinvasion (p < 0.05) and lymph node metastasis (p < 0.005). VEGFR-3 expression was higher in the rectum than in the colon (p < 0.05). VEGF expression correlated with VEGF-D (p < 0.05) and VEGFR-3 (p < 0.005) expression, while VEGF D expression showed no significant association with VEGFR-2 or VEGFR-3. EGFR amplification was present in 10.6% and was not associated with EGFR protein expression. VEGFR-2 and VEGFR-3 expression levels were related to poor patient survival. Stage, perineural invasion, and lymph node metastasis were independent prognostic factors based on a Cox analysis. VEGFR-2 and VEGFR-3 expression are markers of a poor prognosis in patients with surgically resected colorectal adenocarcinoma, whereas EGFR has a minor influence. PMID- 21635553 TI - Genetic polymorphisms of TP53-binding protein 1 (TP53BP1) gene and association with breast cancer risk. AB - In the present study, we evaluated the association between the TP53BP1 Glu353Asp and T-885G polymorphisms and breast cancer risk as well as with the clinicopathological characteristics of the patients. Genotyping of these polymorphisms was performed on 387 breast cancer patients and 252 normal and healthy women who had no history of any malignancy using PCR-RFLP method in a hospital-based Malaysian population. Breast cancer risk was not observed among women who were heterozygous (OR(adj) = 0.887; 95% CI, 0.632-1.245) or homozygous (OR(adj) = 1.083; 95% CI, 0.595-1.969) for Asp allele, and those carriers of Asp allele (OR(adj) = 0.979; 95% CI, 0.771-1.243). Similarly, women who were TG heterozygotes (OR(adj) = 1.181; 95% CI, 0.842-1.658) or GG homozygotes (OR(adj) = 1.362; 95% CI, 0.746-2.486) and carriers of G allele (OR(adj) = 1.147; 95% CI, 0.903-1.458) were not associated with increased risk of breast cancer. Asp allele genotype was significantly associated with ER negativity (p = 0.0015) and poorly differentiated tumours (p = 0.008), but G allele genotype was not associated with the clinicopathological characteristics. In conclusion, Glu353Asp and T-885G polymorphic variants might not have an influence on breast cancer risk, thus might not be potential candidates for cancer susceptibility. Glu353Asp variant might be associated with tumour aggressiveness as defined by its association with ER negativity and poorly differentiated tumours. PMID- 21635554 TI - Sublingual vaccination with sonicated Salmonella proteins and mucosal adjuvant induces mucosal and systemic immunity and protects mice from lethal enteritis. AB - Salmonella enteritidis is one of the most common pathogens of enteritis. Most experimental vaccines against Salmonella infection have been applied through injections. This is a new trial to explore the effect of sublingual administration of Salmonella vaccines on systemic and mucosal immunity. Adult BALB/c mice were sublingually vaccinated with sonicated Salmonella proteins (SSP) alone, or plus adjuvant CpG DNA (CpG) or cholera toxin (CT). They were boosted 2 weeks later. Saliva specific secretory IgA (SIgA) antibody responses were significantly stimulated in the mice vaccinated with SSP only or together with CpG or CT. Whereas the mice sublingually vaccinated with SSP and CpG had higher spleen cell IFN-gamma production and serum specific IgG2a antibody responses, those receiving SSP and CT showed enhanced spleen cell IL-4, IL-5 and IL-6 production, and serum specific IgG1 antibody responses. After oral challenge with live S. enteritidis, the same strain of the source of SSP, immune protection in those sublingually vaccinated with SSP and CpG or CT was found to prevent intestinal necrosis and to render a higher survival rate. In conclusion, sublingual vaccination together with mucosal adjuvant CpG or CT is a simple but effective way against enteric bacterial pathogens. PMID- 21635555 TI - Recurrent meningococcal sepsis in a presumptive immunocompetent host shown to be complement C5 deficient-a case report. AB - Invasive meningococcal disease is a world wide challenge. Most cases occur in immunocompetent children and young adults, but some immunodeficiencies are linked to a greater risk of invasive neisserial infections. One of these is complement component deficiencies, particularly deficiency of properdin and the terminal complement components. We describe a case of recurrent meningococcal sepsis in a young man who was later diagnosed with complete lack of complement component C5. This case report emphasizes the need of having complement deficiencies in mind when being introduced to patients with invasive Neisseria-infections. PMID- 21635556 TI - Importance of vacAs1 gene in gastric cancer patients infected with cagA-negative Helicobacter pylori. PMID- 21635557 TI - The alveolar process of the edentulous maxilla in periodontitis and non periodontitis subjects. AB - BACKGROUND: Early implant failures may document that the bone tissue or the wound healing process following installation surgery was compromised. Subjects who have lost teeth for periodontal reasons exhibit more earlier implant failures than subjects who had experienced tooth loss for other reasons. AIM: To describe the tissue of the fully healed extraction sites in subjects who had lost teeth as a result of periodontitis or for other reasons. MATERIAL AND METHODS: Thirty-six otherwise healthy, partially dentate subjects with fully healed edentulous portions in the posterior maxilla were included. Nineteen of these subjects had lost teeth because of advanced periodontitis (group P) and 17 for other reasons (group NP). Using a trephine drill, a 4-6 mm long hard tissue specimen was harvested. The biopsies were decalcified, embedded in paraffin, sectioned, stained and examined. RESULTS: The edentulous posterior maxilla was comprised of 47.1 +/- 11% lamellar bone, 8.1 +/- 7.1% woven bone, 4.3 +/- 3.1% osteoid and 16.5 +/- 10.4% bone marrow. There were no significant differences in the tissue composition of post-extraction sites of (i) P and NP subjects and (ii) premolar and molar sites. CONCLUSION: More than 50% of the edentulous maxilla was comprised of mineralized bone (lamellar and woven bone). The bone trabeculae frequently appeared to have a random orientation. The direction of the trabeculae rather than the lack of mineralized bone tissue may explain the clinical impression that the bone in the posterior maxilla provides limited resistance to mechanical instrumentation. PMID- 21635558 TI - EAO summer camp: a facilitated sharing experience. AB - AIM: The purpose of the EAO summer camp was to create visions and ideas for future developments in the field of implant dentistry. An additional goal was the installation of a young, strong and enduring network for scientific exchange among participants. SUMMER CAMP ACTIVITY: Forty participants younger than 40 years of age, from 16 different European countries, discussed potential future developments of implant dentistry in a professionally moderated workshop. Participants worked in a competitive manner over 3 days in small teams on four topics: future teaching and education, surgery in 2030, prosthetics in 2030 and futuristic tissue development related to the field of implantology. Various innovative conference and moderation techniques were applied to achieve a maximum output from the creative potential present. RESULTS: Plenum consensus was obtained for several key factors potentially influencing future development in implant dentistry. In particular, teaching and education will be improved by the establishment of curriculum standards and novel teaching technologies. Surgery in 2030 will benefit from an improved cost-effectiveness of new technologies and biomaterials. A more comprehensive knowledge on host susceptibility will have an impact on treatment planning and the predictability of implant therapy. A virtual patient concept and tissue engineering will influence Prosthodontics in 2030. Futuristic tissue development will set a "platinum standard" for tissue regeneration. SUMMARY: Visions on all four topics were generated and discussed intensively during the conference. "Future teaching and education" was voted unanimously as the winning team based on the presented ideas and the special interest this topic generated. PMID- 21635559 TI - Fracture strength of yttria-stabilized tetragonal zirconia polycrystals crowns with different design: an in vitro study. AB - OBJECTIVES: The aim of this study was to evaluate the effect of different types and design of zirconia frameworks as well as the effect of different veneering ceramics on the fracture strength of crowns. The importance of different abutment materials was also evaluated. MATERIALS AND METHODS: Eighty cores, 40 in a fully sintered zirconia material and 40 in a pre-sintered zirconia material were made. Twenty cores of each material were made with a core of even thickness shape (ES) and 20 were made with a core with anatomical shape (AS). The cores were divided into subgroups and veneered with one of two different veneering ceramics: a porcelain and a glass-ceramic material. In total eight groups of 10 crowns were made. They were all cemented onto abutments made of resin. One extra group of 10 AS, pre-sintered zirconia cores veneered with glass-ceramic were made and cemented onto titanium abutments. All crowns underwent thermocycling and mechanical pre-load and were finally loaded until fracture. RESULTS: AS crowns withstood significantly higher loads than ES crowns (P-value <0.001), and crowns with titanium abutments withstood significantly higher loads than crowns supported by abutments made of inlay pattern resin (P-value <0.001). Three types of fracture were noted: minor and major fracture of the veneering ceramic, and complete fracture through core and veneer. ES crowns showed significantly more major fractures of the veneering ceramic than AS crowns. CONCLUSIONS: This in vitro study indicates that the design of the core, as well as the abutment support, significantly influences fracture load and fracture mode of yttria stabilized tetragonal zirconia polycrystals crowns. PMID- 21635560 TI - Does bone mineral density influence the primary stability of dental implants? A systematic review. AB - OBJECTIVE: The aim of this systematic review was to investigate the influence of bone mineral density on the primary stability of dental implants. MATERIAL AND METHODS: A search of health science databases (Cochrane Library, MEDLINE-PubMed, ISI Web of Knowledge, EMBASE, LILACS) and grey literature was performed, including papers published until January 2011. The main key words used were "bone density" (MeSH/DeCS), "dental implant" (MeSH/DeCS), "implant stability", "implant stability quotient", "ISQ", "resonance frequency analysis", "RFA", "Osstell", "Periotest value", "PTV", "Periostest", "insertion torque", "placement torque", "cutting torque". The inclusion criteria comprised observational clinical studies performed in patients who received dental implants for rehabilitation; studies that evaluated the association between bone mineral density and implant primary stability; bone density assessment performed by measurement of Hounsfield units using cone beam computed tomography; and dental implant primary stability evaluated by ISQ value, PTV value or insertion torque measurement. The articles selected were carefully read and classified as low, moderate and high methodological quality, and data of interest were tabulated. RESULTS: Ten articles met the inclusion criteria, but only seven were included because of overlapping patients. They were classified as low or moderate methodological quality and control of bias, and presented positive association between primary stability and bone density. CONCLUSIONS: There is a positive association between implant primary stability and bone mineral density of the receptor site. However, the methodological quality and control of bias of the studies should be improved to produce stronger evidences. PMID- 21635561 TI - The pathopysiology of heat stroke: an integrative view of the final common pathway. AB - Heat stroke is defined as a condition in which body temperature is elevated to such a level that it becomes a noxious agent causing body tissue dysfunction and damage with a characteristic multi-organ clinical and pathological syndrome. Marked hyperthermia, usually above 40.5 degrees C and associated encephalopathy, occurs after thermoregulation is subordinated to circulatory and metabolic demands and to the associated systemic inflammatory reaction. Exertional heat stroke is a function of both intrinsic and extrinsic modulators. Intrinsic modulators like genetics, fitness, acclimatization, illness, medications, and sleep quality can alter individual risk and outcomes, while extrinsic modulators like exercise intensity and duration, clothing and equipment, ambient temperature, relative humidity, and solar radiation can affect the group risk and outcomes. This review integrates the current theoretical and accepted knowledge of physiological alterations into one model that depicts a common pathway from heat stress to heat stroke. PMID- 21635562 TI - Infrapatellar fat pad size, but not patellar alignment, is associated with patellar tendinopathy. AB - Patellar tendinopathy (PT) is one of the most common overuse injuries of the knee. Recent reports indicate that increased body mass is frequently associated with tendinopathy, not only biomechanically but biochemically. Abnormalities of other structures within the knee extensor mechanism [patellofemoral joint (PFJ) alignment and patellar tendon length] that can directly influence the strain distribution of the patellar tendon are inconsistently implicated in PT. The aim of this study was to compare the infrapatellar fat pad volume, patellar tendon length and PFJ alignment in people with chronic PT and a group of age-, gender-, height-, and activity-matched controls with normal tendons. Axial magnetic resonance (MR) images, from 26 participants with PT and 28 control participants were obtained. Fat pad size, patellar tendon length and PFJ alignment were measured digitally from the MR images, using measurement software, and the results compared between the PT group and control group. People with PT had a significantly larger fat pad than healthy controls when controlled for height (P=0.04). Patellar tendon length was not significantly different between groups (P=0.16), nor were there between-group differences for the measures of PFJ alignment (P=0.07-0.76). Thus, the infrapatellar fat pad may play an important role in PT. PMID- 21635563 TI - Receptor dependent immobilization of spermatozoa by sperm immobilization factor isolated from Escherichia coli: proof of evidence. AB - OBJECTIVE: To examine the receptor-ligand interaction between E. coli and spermatozoa resulting in sperm immobilization. METHODS: Sperm immobilization factor (SIF) was isolated and purified from filtrate of E. coli suspension. Receptor on human spermatozoa for SIF was made to isolated and purified by salt extraction, gel permeation and ion exchange chromatography. Receptor dependent immobilization of spermatozoa by SIF was confirmed by competitive inhibition by addition of the purified receptor and binding to sperm receptor by Fluorescin Isothiocynate (FITC) labelled SIF using fluorescence microscopy. RESULTS: Heat labile, non-dialyzable, sperm immobilization factor of ~56 kDa was isolated and purified from E. coli. Using SIF as a tool, receptor of 113 kDa could be isolated and purified from human spermatozoa. Addition of purified receptor completely inhibited sperm immobilization and death induced by SIF suggesting receptor dependent immobilization of spermatozoa. Further incubation of sperm with FITC labelled SIF resulted in staining of whole sperm. CONCLUSION: The study provides evidence of receptor-ligand interaction between E. coli and spermatozoa. PMID- 21635564 TI - Do men have more severe psoriasis than women? PMID- 21635565 TI - Low-dose UVA1 phototherapy for scleroderma: what benefit can we expect? AB - BACKGROUND: The first reports of the application of ultravioletA1 (UVA1) phototherapy for scleroderma go back only to 1995, but since then, several studies have proven its effectiveness in this disease. OBJECTIVES: To evaluate the effectiveness of low-dose (35 J/cm(2) ) UVA(1) phototherapy in patients with scleroderma, trying to define the subgroups that benefit most from this treatment. METHODS: Retrospective analysis of patients diagnosed with clinical and/or histological scleroderma, undergoing low-dose UVA(1) phototherapy between 2003 and 2010. RESULTS: We studied 21 patients (20 women and one man) aged 10-75 years (mean 50). Eighteen with morphea performed a total of 29 treatments, with an average of 33 sessions per treatment per patient, a mean dose of 31 J/cm(2) per session and an average cumulative dose of 1662 J/cm(2) (310-4270). The three patients with systemic scleroderma underwent a total of five treatments, with an average of 26 sessions per treatment per patient, a mean dose of 29.5 J/cm(2) per session and an average cumulative dose of 1160 J/cm(2) (660-1695). Using the modified Rodnan skin score, in the group of patients with morphea, a marked improvement was found in 77.8% patients and a moderate improvement was found in 11.1% patients. In the systemic scleroderma group, a patient with complete remission of the skin sclerosis is emphasized. CONCLUSIONS: Our experience allows us to conclude that low-dose UVA(1) phototherapy is a well tolerated option, with excellent results mainly in patients with morphea. This treatment is a valuable contribution to these patients, given the limited therapeutic options available. PMID- 21635566 TI - Contribution of the P2X7 1513A/C loss-of-function polymorphism to extrapulmonary tuberculosis susceptibility in Tunisian populations. AB - The P2X7 receptor has been found to be linked to an increased risk for tuberculosis in some populations. In this study, we investigate whether the P2X7 receptor plays a role in increasing susceptibility to tuberculosis in Tunisia. We examined two 1513A/C and -762T/C polymorphisms at the P2X7 receptor in 168 patients with pulmonary TB (pTB), 55 patients with extrapulmonary TB (epTB) and 150 blood donors from Tunisia. Genotyping of 1513A/C and -762T/C polymorphisms was performed in purified genomic DNA using PCR-restriction fragment length polymorphism and allele-specific PCR, respectively. The 1513C, CC and AC loss-of function allele and genotypes were overrepresented in the epTB group compared with the control group (45% vs. 17%, P=10(-8) ; 24% vs. 4%, P=3 * 10(-7) ; 42% vs. 27%, P=10(-3) , respectively). Additionally, they were associated with 3.83-, 11.86- and 3.15-fold risks of developing this clinical tuberculosis form, respectively. No associations between the -762T/C polymorphism and tuberculosis disease, as well as disease anatomic location were observed. Collectively, our results suggest that the P2X7 1513A/C loss-of-function polymorphism may contribute to susceptibility to epTB in Tunisian populations. PMID- 21635567 TI - Characterization of sugar recognition by the toxin complex produced by the Clostridium botulinum serotype C variant strain Yoichi. AB - Clostridium botulinum serotype C strains produce a neurotoxin (BoNT) along with nontoxic proteins, including nontoxic nonhemagglutinin and three hemagglutinin subcomponents, HA-70, HA-33 and HA-17, to form a large toxin complex (L-TC). While L-TCs produced by serotype C strains usually exhibit hemagglutination (HA) activity via HA-33 binding to sialic acid on erythrocytes, serotype C strain Yoichi (C-Yoichi) L-TC exhibited neither HA nor binding activity towards erythrocytes, probably due to a C-terminal truncation of the HA-33 protein. However, here, we demonstrate that C-Yoichi L-TC newly showed full HA and binding activity towards neuraminidase-treated erythrocytes that was completely inhibited in the presence of galactose (Gal) or lactose (Lac). Binding of C-Yoichi L-TC to rat small intestine epithelial cells (IEC-6) treated with neuraminidase was also significantly enhanced compared with untreated IEC-6 cells. Similarly, the HA 33/HA-17 complex isolated from C-Yoichi L-TC also bound to neuraminidase-treated IEC-6 cells. The binding activity of both L-TC and HA-33/HA-17 was inhibited in the presence of Gal or Lac. Additionally, C-Yoichi L-TC adsorbed tightly to a lactose-affinity gel column. These results strongly suggest that the unusual recognition of the Gal moiety on the cells could be due to a variation and/or a truncation in the C-terminal-half of the unique C-Yoichi HA-33 protein. PMID- 21635568 TI - The relative efficacy of different strain combinations of lactic acid bacteria in the reduction of populations of Salmonella enterica Typhimurium in the livers and spleens of mice. AB - Multispecies probiotics have been reported to be more effective than monostrain probiotics in health promoting for the host. In this study, 12 lactic acid bacteria (LAB) strains were selected based on the level of induction of tumor necrosis factor (TNF)-alpha in RAW 264.7 macrophage cells. Their adherence to Caco-2 cells and inhibitory effects on Salmonella invasion of Caco-2 cells were compared. Strains with different probiotic properties were then combined and BALB/c mice were fed with LAB strains for 63 days; then the mice were challenged with Salmonella on day 64. For Salmonella-unchallenged mice that received a multistrain combination of LAB strains that have greater TNF-alpha production in macrophages, greater adherence and inhibit Salmonella invasion of Caco-2 cells to a greater extent, their peritoneal macrophages had greater phagocytic activity. For Salmonella-challenged mice, a significant reduction of Salmonella cells in the livers and spleens of the mice was observed 8 days post challenge. The addition of 12% skim milk powder together with LAB strain combinations significantly enhanced the reduction of Salmonella cells in the mice livers and spleens. In conclusion, we have shown that LAB strain combinations with particular probiotic properties when fed to mice can inhibit Salmonella invasion of the liver and spleen. PMID- 21635569 TI - Acylated cholesteryl galactosides are ubiquitous glycolipid antigens among Borrelia burgdorferi sensu lato. AB - Lyme disease (LD) is the most common tick-borne disease in the Northern hemisphere. It is caused by Borrelia burgdorferi sensu lato, in particular, B. burgdorferi sensu stricto, Borrelia garinii, and Borrelia afzelii. However, other genospecies have been implicated as causative factors of LD as well. Borrelia burgdorferi exhibits numerous immunogenic lipoproteins, but due to strong heterogeneity, the use of these proteins for serodiagnosis and vaccination is hampered. We and others have identified acylated cholesteryl galactosides (ACGal) as a novel glycolipid present in B. burgdorferi sensu stricto, B. afzelii, and B. garinii. ACGal is a strong antigen and the majority of patients display anti ACGal antibodies in the chronic stages of LD. However, it is unknown whether ACGal is present in other presumably pathogenic B. burgdorferi genospecies. Therefore, we performed an analysis of the total lipid extracts of a wide spectrum of genospecies of B. burgdorferi sensu lato using thin-layer chromatography as well as Western blot and dot-blot assays. We show that ACGal is present in substantial quantities in all B. burgdorferi genospecies tested. Therefore, this molecule might improve the serological detection of rarely pathogenic genospecies, and may be used as a protective vaccine regardless of the prevailing genospecies. PMID- 21635570 TI - Rapid detection of hepatitis C virus RNA by a reverse transcription loop-mediated isothermal amplification assay. AB - The usefulness of reverse transcription loop-mediated isothermal amplification (RT-LAMP) for the rapid diagnosis of hepatitis C virus (HCV) RNA was evaluated. This assay showed higher sensitivities than that of nested RT-PCR, with a detection limit of 600 IU mL(-1) , and no cross-reactivity was observed with hepatitis A virus, hepatitis B virus and hepatitis E virus. Furthermore, 106 stored sera from recently diagnosed cases were retrospectively investigated with real-time RT-PCR, the nested RT-PCR, in parallel with this new assay. The general detection rates of HCV RT-LAMP, real-time PCR and the nested RT-PCR for 106 stored sera samples were 95%, 96% and 88%, respectively. This study provides the first data on the usefulness of HCV RT-LAMP in the diagnosis of HCV RNA, especially in the early clinical diagnosis of acute HCV infection. PMID- 21635571 TI - Effects of diets with different protein and DL-methionine levels on the growth performance and N-balance of growing minks. AB - This study was performed to evaluate the effects of diets with different protein and dl-methionine (Met) levels on nitrogen (N) retention, nutrient digestibility, growth performance, and some blood parameters in growing minks. Eighty healthy male minks were selected and randomly divided into five groups with different types of diet. The dietary protein levels, expressed as percentage of dry matter (DM), were 36% (HP) and 28% (LP), corresponding to average 363g and 295g protein/kg DM, respectively. LP was supplemented with Met (0.3%, 0.6% and 0.9% DM); the codes were LP+M1, LP+M2 and LP+M3, respectively. From July to middle of September, the average daily gain and feed: gain ratio (F/G) of the minks that received the diet with 0.6% Met added to the low protein diet was better than feeding HP and other groups. Fecal N and Urinary N of group LP+M2 were the lowest one, in contrast, the daily retention of N was the highest one. Digestibility of DM and CP were not affected by different diets, but digestibility of fat declined with dietary protein level decreasing. Serum urea nitrogen (SUN) was affected by different protein and Met levels. Considering all factors the best performance could be observed offering LP+M2, the prime level of Met was 13.87 g/kg DM in dietary, and 258.5 g digestible protein kg(-1) DM was enough for mink in growing period. Furthermore, addition of Met in low protein diets for minks would be beneficial in terms of reduced feed expenses and lower nitrogen emissions to the environment. PMID- 21635572 TI - Dry matter and calcium digestibility in captive veiled chameleons (Chamaeleo calyptratus). AB - Although metabolic bone disease (MBD) is a very common disease in reptiles kept as pets, empirical data on the calcium (Ca) metabolism of reptiles are still scarce. We used the opportunity of a large-scale experimental study on growth and clinical manifestations of MBD in captive veiled chameleons (Chamaeleo calyptratus) to measure the apparent dry matter (DM) and Ca digestibility in 19 animals (6-49 g), receiving locust nymphs (Locusta migratoria) of two size classes (0.05 and 0.5 g) with or without supplementation of Ca, vitamin A and cholecalciferol (Group A: Ca 0.04-0.09%DM; Group B: Ca 0.47-0.84%DM). Dry matter digestibility was significantly lower for animals receiving smaller-sized prey. A regression analysis of dietary Ca vs. digestible Ca content revealed a complete 'true' digestibility of Ca for the range of investigated diets, which might indicate that requirements for this mineral were not yet exceeded by the diets used (so that a reduction in Ca absorption would be induced). Options of higher dietary Ca provision, and reactions of chameleons to such diets, should be further investigated. PMID- 21635573 TI - Effects of dietary carbohydrate sources on plasma glucose, insulin and IGF-I levels in multiparous sows. AB - Effects of different carbohydrate sources on plasma glucose, insulin and insulin like growth factor-I (IGF-I) levels were compared to subsequently be able to study effects of insulin-stimulating diets on follicle development in sows. The following feed components were tested in 12 sows during six consecutive test periods of 9.5 days: dextrose (DEX), sucrose (SUC), lactose (LAC), dextrose plus lactose (DL), sucrose plus lactose (SL), dextrose plus sugarbeet pulp (DSBP) and control (CON). On day 2, 5 and 9 of each test period, plasma glucose (only at day 9), insulin and IGF-I profiles were determined. Despite similar glucose profiles for all diets, the postprandial insulin response was higher for DL and SL compared with CON and LAC; the other diets were intermediate. Plasma IGF-I levels were higher in CON, LAC and SL compared with DSBP, but differences were only marginal. It was concluded that dextrose and sucrose have the potential to stimulate fast and high insulin peaks, especially when combined with additional lactose. Despite the high dextrose in the DSBP diet, the insulin response was flattened, probably due to the viscosity of sugarbeet pulp. The results show that modulation of plasma insulin levels by dietary carbohydrates seems possible in anabolic sows, but IGF-I levels are less easily modified. PMID- 21635574 TI - Meta-analysis of the relationship between dietary tannin level and methane formation in ruminants from in vivo and in vitro experiments. AB - A meta-analysis was conducted to evaluate the extent to which dietary tannin level is related to methane emissions from ruminants. Data from a total of 30 experiments comprising 171 treatments were entered in a database. In vitro batch culture and in vivo measurements were distinguished as experimental approaches. With any approach, methane declined when dietary tannins increased. The in vitro approach predicted the in vivo response quite accurately. However, in vitro, the response followed a quadratic response pattern (R(2) = 0.66; lower response with increasing tannin level), whereas in vivo, this decline was linear (R(2) = 0.29). This indicates that the in vitro batch culture is of limited accuracy for estimating effects at levels >100 g tannin/kg dry matter. The large variation in methane/digestible organic matter (OM) found at low tannin levels may explain contrasting literature reports. Methane reduction with tannins was associated with a reduced apparent digestion of OM, and especially fibre, but methane/apparently digestible OM declined also. The present findings are helpful as they identified an underlying general antimethanogenic effect of tannins across tannin sources and experimental conditions, thus allowing concentrating the search on sources with satisfactory palatability and low adverse effects on animal performance. PMID- 21635575 TI - Effect of feeding Bacillus subtilis natto fermentation product on milk production and composition, blood metabolites and rumen fermentation in early lactation dairy cows. AB - This experiment was conducted to determine the effect of Bacillus subtilis natto fermentation product supplementation on blood metabolites, rumen fermentation and milk production and composition in early lactation dairy cows. Thirty-six multiparous Holstein cows (DIM = 29 +/- 6 days, parity = 2.8 +/- 1.1) were blocked by DIM and parity and then randomly assigned to three treatments (12 per treatment) in a 9-week trial. Cows in control, DFM1 and DFM2 were fed TMR diets supplemented with 0, 6 and 12 g of B. subtilis natto solid-state fermentation product per day per cow respectively. Plasma non-esterified fatty acids were lower (p = 0.03) in DFM1 and DFM2 compared with control cows (633 and 639 vs. 685 MUm). Ruminal propionate increased (23.9 vs. 26.3 and 26.9/100 mol, control vs. DFM1 and DFM2 respectively) and acetate decreased (64.2 vs. 62.7 and 62.1/100 mol, control vs. DFM1 and DFM2 respectively) with increasing B. subtilis natto fermentation product supplementation. DMI of the cows in three treatments was not affected by B. subtilis natto fermentation product supplementation, but milk yield was 3.1 and 3.2 kg/day higher for DFM1 and DFM2 than that for control cows on average across the 9-week trial, and significant differences were observed during weeks 5-9 of the trial, which resulted in 9.5% and 11.7% increase in feed efficiency. B. subtilis natto fermentation product supplementation did not affect milk fat percentage and protein yield but increased (p < 0.05) milk fat yield and lactose percentage (p < 0.01) and tended to decrease protein percentage (p = 0.06). The findings show that B. subtilis natto fermentation product was effective in increasing lactation performance of early lactation dairy cows possibly by altering the rumen fermentation pattern without any negative effects on blood metabolites. PMID- 21635576 TI - Effect of glucose addition and N sources in defined media on fibrolytic activity profiles of Neocallimastix sp. YQ1 grown on corn stover. AB - Cleavage of plant cell wall arabinoxylans occurs by the action of ferulic acid esterase (FAE) and acetyl esterase (AE), which cleave feruloyl groups substituted at the 5'-OH group of arabinosyl residues and acetyl groups substituted at O-2/O 3 of the xylan backbone, respectively. In this study, we examined the enzyme profiles of the anaerobic rumen fungus Neocallimastix sp. YQ1 for FAE, AE and polysaccharide hydrolases when grown on corn stover, a lignin-rich waste biomaterial. A 2 * 4 factorial experiment in 10-days pure cultures was used to test glucose addition (G(+) : glucose at 1.0 g/l, G(-) : no glucose) and four N sources (N1: 1.0 g/l yeast extract, 1.0 g/l tryptone and 0.5 g/l (NH(4))(2) SO(4); N2: 2.8 g/l yeast extract and 0.5 g/l (NH(4))(2) SO(4) ; N3: 1.6 g/l tryptone and 0.5 g/l (NH(4))(2) SO(4); N4: 1.4 g/l tryptone and 1.7 g/l yeast extract) in defined media. The optimal combinations of glucose and N sources to promote FAE and AE activity were G(+) N2 and G(+) N4, respectively. The peak activities of FAE and AE occurred on days 9 and 10, respectively. Addition of glucose and an increase in yeast extract and/or tryptone to a Hungate's medium favoured fungal production of volatile fatty acids, which could be just a consequence of more organic matter available to digest. This suggests that enzymatic release of ferulic acid by a synergistic action of lignin hydrolytic esterase and polysaccharide hydrolases may be essential for plant cell wall biodegradation in the rumen. PMID- 21635577 TI - Differential effects of propionate or beta-hydroxybutyrate on genes related to energy balance and insulin sensitivity in bovine white adipose tissue explants from a subcutaneous and a visceral depot. AB - Ruminants rely on short-chain fatty acids (SCFA) as principal energy source. Herein, we compared the effects of propionate, beta-hydroxybutyrate (BHB) and insulin on mRNA abundance of energy balance-related genes by short-term incubation (4 h) in bovine subcutaneous (SC) and retroperitoneal (RP) adipose tissue (AT) explants in vitro. Propionate either significantly (p < 0.05), or as a trend (p <= 0.1) affected mRNA abundance of genes such as adiponectin system in both depots in treated samples versus controls. Propionate increased adiponectin receptor 1 (AdipoR1) and AdipoR2 mRNA only in SC AT. beta-hydroxybutyrate decreased mRNA abundance of adiponectin and AdipoR1 in SC AT as a trend. The mRNA abundance of free fatty acid receptor 2/3 (FFAR2/3) and other genes of interest (GOI) was increased during differentiation in bovine preadipocyte culture. The mRNA abundance of all the GOI remained unchanged after short-term insulin stimulation. In total, propionate, BHB or insulin during short-term treatment exert divergent effects on the mRNA abundance of GOI in both depots in vitro. Our results indicate that the bovine adiponectin system might be more sensitive to propionate than to BHB. We demonstrated the presence of FFAR2/3 mRNA not only in both AT depots but also in differentiating preadipocytes isolated from bovine SC AT. Therefore, we established that SCFA are able to exert insulin-independent effects on bovine adipose tissue, which might be independent from propionate uptake-related events. PMID- 21635578 TI - Effect of linoleic acid albumin in a dilution solution and long-term equilibration for freezing of bovine spermatozoa with poor freezability. AB - Despite normal eucrasia, mating desire and semen quality, sire bulls sometimes have spermatozoa with poor freezing tolerance. This study assessed effects of the addition of linoleic acid albumin (LAA) and long-term (LT) equilibrium to frozen semen on their sperm freezing tolerance. Immediately after collection using an artificial vagina and a breeding mount, semen was diluted with yolk citrate buffer; then, it was cooled slowly to 4 degrees C during more than 5 h. Equilibrium treatment at 4 degrees C was applied using the same extender supplemented with glycerol. Semen of bull A, with low sperm freezing tolerance, was treated with 1 mg/ml of LAA added to the first extender. The equilibrium treatment at 4 degrees C was prolonged to 30 h. Significantly higher motility rates were obtained for the LT + LAA-treated sperm before and after freezing thawing. However, for semen of bulls B and C with normal sperm freezing tolerance, the LT + LAA treatment barely exhibited a small effect on the motility rate. Almost no difference was found among bulls A, B and C in the motility rates of LT + LAA-treated sperm after freezing-thawing. No difference of fertility was apparent on LT + LAA-treated frozen sperm in comparison with normal sperm in embryonic collection and in vitro fertilization. It was not an aberration of fertility in vivo or in vitro. In addition, the conception rate of artificial insemination did not have a difference, and a normal calf was obtained. Results show that addition of LAA to an extender for frozen bovine spermatozoa and 30 h of low-temperature equilibrium might improve the motility of freezing-thawing spermatozoa with poor freezability. Sperm exhibited normal fertilization capability and ontogenic capability. PMID- 21635579 TI - Effect of freezing on sperm nuclear DNA. AB - Sperm DNA damage has a significant impact on reproductive outcomes. In recent years, the search for optimal molecular markers for the evaluation of semen quality has resulted in the increased focus on sperm nuclear DNA assessment. The primary aim of this article was to review and summarize the effects of freezing thawing procedure on nuclear DNA integrity of boar spermatozoa. Using different sperm DNA integrity assays, it has been confirmed that the sperm DNA undergoes structural changes during the freezing-thawing process. Evidence has been shown that a significant proportion of frozen-thawed spermatozoa with compromised chromatin integrity was highly susceptible to DNA fragmentation. Moreover, the possible mechanisms responsible for post-thaw sperm DNA damage could be because of cryo-induced oxidative stress and, to a lesser extent, to the activation of an apoptotic-like phenomenon. This review also highlights the ongoing effort employed to develop optimal strategies to reduce sperm DNA damage following freezing-thawing of boar semen. PMID- 21635580 TI - The pathogenic persona of community-associated oral streptococci. AB - The mitis group streptococci (MGS) are widespread in the oral cavity and are traditionally associated with oral health. However, these organisms have many attributes that contribute to the development of pathogenic oral communities. MGS adhere rapidly to saliva-coated tooth surfaces, thereby providing an attachment substratum for more overtly pathogenic organisms such as Porphyromonas gingivalis, and the two species assemble into heterotypic communities. Close physical association facilitates physiologic support, and pathogens such as Aggregatibacter actinomycetemcomitans display resource partitioning to favour carbon sources generated by streptococcal metabolism. MGS exchange information with community members through a number of interspecies signalling systems including AI-2 and contact dependent mechanisms. Signal transduction systems induced in P. gingivalis are based on protein dephosphorylation mediated by the tyrosine phosphatase Ltp1, and converge on a LuxR-family transcriptional regulator, CdhR. Phenotypic responses in P. gingivalis include regulation of hemin uptake systems and gingipain activity, processes that are intimately linked to the virulence of the organism. Furthermore, communities of S. gordonii with P. gingivalis or with A. actinomycetemcomitans are more pathogenic in animal models than the constituent species alone. We propose that MGS should be considered accessory pathogens, organisms whose pathogenic potential only becomes evident in the context of a heterotypic microbial community. PMID- 21635581 TI - Heterologous protein transfer within structured myxobacteria biofilms. AB - Microbial biofilms represent heterogeneous populations of cells that form intimate contacts. Within these populations cells communicate, cooperate and compete. Myxobacteria are noted for their complex social interactions, including gliding motility and lipoprotein exchange. Here, we investigated cis protein sequence and cellular behaviour requirements for lipoprotein transfer between Myxococcus xanthus cells. Specifically, an outer membrane (OM) type II signal sequence (SS) fused to the heterologous mCherry fluorescent reporter resulted in OM localization. When donor cells harbouring SS(OM)-mCherry were mixed with GFP labelled recipient cells they developed red fluorescence. Our results surprisingly showed that a type II SS for OM localization, but not inner membrane localization, was necessary and sufficient for rapid and efficient heterologous protein transfer. Importantly, transfer did not occur in liquid or on surfaces where cells were poorly aligned. We conclude that cell-cell contact and alignment is a critical step for lipoprotein exchange. We hypothesize that protein transfer facilitates cooperative myxobacteria behaviours. PMID- 21635582 TI - The translational regulatory function of SecM requires the precise timing of membrane targeting. AB - In Escherichia coli, secA expression is regulated at the translational level by an upstream gene (secM) that encodes a presecretory protein. SecM contains a C terminal sequence motif that induces a transient translation arrest. Inhibition of SecM membrane targeting prolongs the translation arrest and increases SecA synthesis by concomitantly altering the structure of the secM-secA mRNA. Here we show that the SecM signal peptide plays an essential role in this regulatory process by acting as a molecular timer that co-ordinates membrane targeting with the synthesis of the arrest motif. We found that signal peptide mutations that alter targeting kinetics and insertions or deletions that change the distance between the SecM signal peptide and the arrest motif perturb the balance between the onset and release of arrest that is required to regulate SecA synthesis. Furthermore, we found that the strength of the interaction between the ribosome and the SecM arrest motif is calibrated to ensure the release of arrest upon membrane targeting. Our results strongly suggest that several distinctive features of the SecM protein evolved as a consequence of constraints imposed by the ribosome and the Sec machinery. PMID- 21635583 TI - The safety of flecainide treatment of atrial fibrillation: long-term incidence of sudden cardiac death and proarrhythmic events. AB - OBJECTIVE: To assess the safety of long-term treatment with flecainide in patients with atrial fibrillation (AF), particularly with regard to sudden cardiac death (SCD) and proarrhythmic events. DESIGN: Retrospective, observational cohort study. SETTING: Single-centre study at Orebro University Hospital, Sweden. Subjects. A total of 112 patients with paroxysmal (51%) or persistent (49%) AF (mean age 60 +/- 11 years) were included after identifying all patients with AF who initiated oral flecainide treatment (mean dose 203 +/- 43 mg per day) between 1998 and 2006. Standard exclusion/inclusion criteria for flecainide were used, and flecainide treatment was usually combined with an atrioventricular-blocking agent (89%). MAIN OUTCOME MEASURE: Death was classified as sudden or nonsudden according to standard definitions. Proarrhythmia was defined as cardiac syncope or life-threatening arrhythmia. RESULTS: Eight deaths were reported during a mean follow-up of 3.4 +/- 2.4 years. Compared to the general population, the standardized mortality ratios were 1.57 (95% confidence interval (CI) 0.68-3.09) for all-cause mortality and 4.16 (95% CI 1.53-9.06) for death from cardiovascular disease. Three deaths were classified as SCDs. Proarrhythmic events occurred in six patients (two each with wide QRS tachycardia, 1 : 1 conducted atrial flutter and syncope during exercise). CONCLUSION: We found an increased incidence of SCD or proarrhythmic events in this real-world study of flecainide used for the treatment of AF. The findings suggest that further investigation into the safety of flecainide for the treatment of patients with AF is warranted. PMID- 21635584 TI - Adiponectin and raised mortality in type 1 diabetes: any credible explanatory mechanisms? PMID- 21635585 TI - Tn5 transposase-assisted transformation of indica rice. AB - Here, we describe experiments on Tn5 transposase-assisted transformation of indica rice. Transposomes were formed in vitro as a result of hyperactive Tn5 transposase complexing with a transposon that contained a 19-bp tetracycline operator (tetO) sequence. To form modified projectiles for transformation, the Tn10-derived prokaryotic tetracycline repressor (TetR) proteins, which can bind transposomes via the high affinity of TetR for tetO, were immobilized onto the surface of bare gold microscopic particles. These projectiles were introduced into cells of the indica rice cultivar Zhuxian B by particle bombardment. Once projectiles were inside the cell, tetracycline induced an allosteric conformational change in TetR that resulted in the dissociation of TetR from tetO, and thus generated free transposomes. Molecular evidence of transposition was obtained by the cloning of insertion sites from many transgenic plants. We also demonstrated that the introduced foreign DNA was inherited stably over several generations. This technique is a promising transformation method for other plant species as it is species independent. PMID- 21635586 TI - Knockdown of CENH3 in Arabidopsis reduces mitotic divisions and causes sterility by disturbed meiotic chromosome segregation. AB - The histone H3 variant (CENH3) of centromeric nucleosomes is essential for kinetochore assembly and thus for chromosome segregation in eukaryotes. The mechanism(s) that determine centromere identity, assembly and maintenance of kinetochores are still poorly understood. Although the role of CENH3 during mitosis has been studied in several organisms, little is known about its meiotic function. We show that RNAi-mediated CENH3 knockdown in Arabidopsis thaliana caused dwarfism as the result of a reduced number of mitotic divisions. The remaining mitotic divisions appeared to be error-free. CENH3 RNAi transformants had reduced fertility because of frequently disturbed meiotic chromosome segregation. N-terminally truncated EYFP-CENH3(C) is deposited to and functional within Arabidopsis centromeres of mitotic chromosomes, but cannot be loaded onto centromeres of meiotic nuclei. Thus the N-terminal part is apparently required for CENH3 loading during meiosis. EYFP-CENH3(C) expression reduces the amount of endogenous CENH3, thus mimicking the effect of RNAi. The consequences of reduced endogenous CENH3 and lack of meiotic incorporation of EYFP-CENH3(C) are reduced fertility caused by insufficient CENH3 loading to the centromeres of meiotic chromosomes, subsequent lagging of chromosomes and formation of micronuclei. PMID- 21635587 TI - Forecasting the ward climate: a study from a dementia care unit. AB - AIMS AND OBJECTIVES: This article present findings from a study aiming to explore the psychosocial climate and its influence on the well-being of people with dementia in a psycho-geriatric hospital unit. BACKGROUND: Environmental influence in dementia is well explored in relation to the physical environment; however, few studies have explored the psychosocial environment and its influence on well being. DESIGN: The study had a grounded theory design. METHODS: Participant observations were conducted in a psycho-geriatric ward for assessment and treatment of people with dementia in Sweden (n = 36 hours). Data were collected and analysed in a dialectical fashion using the principles of grounded theory methodology. RESULTS: The basic social process that best accounted for the variation in the psychosocial climate and well-being of patients at the unit was 'staff presence or absence', conceptualised as the core category. Three categories emerged in relation to the core category; 'sharing place and moment', 'sharing place but not moment' and 'sharing neither place nor moment'. CONCLUSIONS: Staff were catalysts for the psychosocial climate and when being present and engaged they could create a climate interpreted as at-homeness which supported patient well-being. When being absent, the climate quickly became anxious and this facilitated patient ill-being. To provide quality care for people with dementia staff need to be aware of their role in setting the emotional tone of the psychosocial climate and also that this emotional tone significantly influences patient well-being. RELEVANCE TO CLINICAL PRACTICE: The findings are clinically relevant and can be operationalised and applied in clinical practice. Awareness of the intimate connection between staff presence and absence, the psychosocial climate and patient well-being highlights an ethical responsibility to question: routines that promote staff absence; a culture of merely 'doing for'; and nursing tasks which involve a minimum of staff patient interaction. The findings have implications for managers as well as for clinical staff. PMID- 21635588 TI - Can food be addictive? Public health and policy implications. AB - AIMS: Data suggest that hyperpalatable foods may be capable of triggering an addictive process. Although the addictive potential of foods continues to be debated, important lessons learned in reducing the health and economic consequences of drug addiction may be especially useful in combating food-related problems. METHODS: In the current paper, we review the potential application of policy and public health approaches that have been effective in reducing the impact of addictive substances to food-related problems. RESULTS: Corporate responsibility, public health approaches, environmental change and global efforts all warrant strong consideration in reducing obesity and diet-related disease. CONCLUSIONS: Although there exist important differences between foods and addictive drugs, ignoring analogous neural and behavioral effects of foods and drugs of abuse may result in increased food-related disease and associated social and economic burdens. Public health interventions that have been effective in reducing the impact of addictive drugs may have a role in targeting obesity and related diseases. PMID- 21635589 TI - Obesity - is food addiction to blame? PMID- 21635590 TI - Food and addiction - sugars, fats and hedonic overeating. PMID- 21635591 TI - Maternal diet and offspring development. PMID- 21635592 TI - Food addiction not helpful: the hedonic component - implicit wanting - is important. PMID- 21635593 TI - All foods are habit-forming - what I want to know is which will kill me! PMID- 21635595 TI - Commentary on Voas et al. (2011): drug-free roadways - can we get there from here? Steps to reducing drugged driving in the United States. PMID- 21635596 TI - Commentary on Makelaet al. (2011): how many patients must be asked about alcohol before it is enough? PMID- 21635597 TI - Comment on Halpern et al. (2011). PMID- 21635598 TI - 'Ecstasy use, by itself, does not result in residual neurotoxicity'- a powerful argument? PMID- 21635600 TI - Bruce Rounsaville (1949-2011). PMID- 21635601 TI - Residual neurocognitive features of ecstasy use: a re-interpretation of Halpern et al. (2011) consistent with serotonergic neurotoxicity. PMID- 21635602 TI - Heterozygosity is unrelated to adult fitness measures in a large, noninbred population of great tits (Parus major). AB - The extent to which heterozygosity-fitness correlations (HFCs) are expected in wild populations is an important and unresolved question in evolutionary biology, because it relates to our understanding of the genetic architecture of fitness. Here, we report a study of HFCs in a wild, noninbred population of great tits (Parus major), based on a sample comprising 281 individuals typed at 26 markers, resulting in a data set comprising over 5600 genotypes. We regressed pedigree derived f-score and multilocus genetic diversity against eight life-history traits known to be associated with fitness in this population, including lifetime reproductive success (LRS), as well as several morphological traits under weak selection. We found no evidence for either multilocus or single-locus HFCs for any morphological or fitness trait, and further found no evidence that effect sizes were stronger for those life-history traits more closely associated with reproductive fitness. This result may, in part, be explained by the fact that we found no evidence that our set of 26 markers had any power to infer genome-wide heterozygosity in this population and that marker-derived heterozygosity was uncorrelated with pedigree-derived f-score. Overall, these results emphasize the fact that the often-reported strong HFCs detected in small, inbred populations do not reflect a general phenomenon of increasing individual reproductive fitness with increasing heterozygosity. PMID- 21635603 TI - Path analysis of the genetic integration of traits in the sand cricket: a novel use of BLUPs. AB - This study combines path analysis with quantitative genetics to analyse a key life history trade-off in the cricket, Gryllus firmus. We develop a path model connecting five traits associated with the trade-off between flight capability and reproduction and test this model using phenotypic data and estimates of breeding values (best linear unbiased predictors) from a half-sibling experiment. Strong support by both types of data validates our causal model and indicates concordance between the phenotypic and genetic expression of the trade-off. Comparisons of the trade-off between sexes and wing morphs reveal that these discrete phenotypes are not genetically independent and that the evolutionary trajectories of the two wing morphs are more tightly constrained to covary than those of the two sexes. Our results illustrate the benefits of combining a quantitative genetic analysis, which examines statistical correlations between traits, with a path model that focuses upon the causal components of variation. PMID- 21635604 TI - Multiple quantitative trait loci influence intra-specific variation in genital morphology between phylogenetically distinct lines of Drosophila montana. AB - The evolution of animal genitalia has gained renewed interest because of their potential roles during sexual selection and early stages of species formation. Although central to understanding the evolutionary process, knowledge of the genetic basis of natural variation in genital morphology is limited to a very few species. Using an outbred cross between phylogenetically distinct lines of Drosophila montana, we characterized quantitative trait loci (QTLs) affecting the size and shape of the distiphallus, a prominent part of the male intromittent organ. Our microsatellite-based linkage analysis shows that intra-specific variation in the distiphallus involves several QTLs of largely additive effect and that a highly significant QTL co-localizes with the same inversion where we have earlier localized a large QTL for a sexually selected courtship song trait. The latter indicates that inversions can play an important role in shaping the evolution of rapidly evolving traits with a potential influence on speciation. PMID- 21635605 TI - The opportunity for sexual selection: not mismeasured, just misunderstood. AB - Evolutionary biologists have developed several indices, such as selection gradients (beta) and the opportunity for sexual selection (I(s) ), to quantify the actual and/or potential strength of sexual selection acting in natural or experimental populations. In a recent paper, Klug et al. (J. Evol. Biol.23, 2010, 447) contend that selection gradients are the only legitimate metric for quantifying sexual selection. They argue that I(s) and similar mating-system based metrics provide unpredictable results, which may be uncorrelated with selection acting on a trait, and should therefore be abandoned. We find this view short-sighted and argue that the choice of metric should be governed by the research question at hand. We describe insights that measures such as the opportunity for selection can provide and also argue that Klug et al. have overstated the problems with this approach while glossing over similar issues with the interpretation of selection gradients. While no metric perfectly characterizes sexual selection in all circumstances, thoughtful application of existing measures has been and continues to be informative in evolutionary studies. PMID- 21635606 TI - Temperature niche shift observed in a Lepidoptera population under allochronic divergence. AB - A process of adaptive divergence for tolerance to high temperatures was identified using a rare model system, consisting of two sympatric populations of a Lepidoptera (Thaumetopoea pityocampa) with different life cycle timings, a 'mutant' population with summer larval development, Leiria SP, and the founder natural population, having winter larval development, Leiria WP. A third, allopatric population (Bordeaux WP) was also studied. First and second instar larvae were experimentally exposed to daily-cycles of heat treatment reaching maximum values of 36, 38, 40 and 42 degrees C; control groups placed at 25 degrees C. A lethal temperature effect was only significant at 42 degrees C, for Leiria SP, whereas all temperatures tested had a significant negative effect upon Leiria WP, thus indicating an upper threshold of survival c.a. 6 degrees C above that of the WP. Cox regression model, for pooled heat treatments, predicted mortality hazard to increase for Leiria WP (+108%) and Bordeaux WP (+78%) in contrast to Leiria SP; to increase by 24% for each additional degrees C; and to decrease by 53% from first to second instar larvae. High variability among individuals was observed, a population characteristic that may favour selection and consequent adaptation. Present findings provide an example of ecological differentiation, following a process of allochronic divergence. Results further contribute to a better understanding of the implications of climate change for ecological genetics. PMID- 21635607 TI - Fitness-dependent mutation rates in finite populations. AB - Mutation rate may be condition dependent, whereby individuals in poor condition, perhaps from high mutation load, have higher mutation rates than individuals in good condition. Agrawal (J. Evol. Biol.15, 2002, 1004) explored the basic properties of fitness-dependent mutation rate (FDMR) in infinite populations and reported some heuristic results for finite populations. The key parameter governing how infinite populations evolve under FDMR is the curvature (k) of the relationship between fitness and mutation rate. We extend Agrawal's analysis to finite populations and consider dominance and epistasis. In finite populations, the probability of long-term existence depends on k. In sexual populations, positive curvature leads to low equilibrium mutation rate, whereas negative curvature results in high mutation rate. In asexual populations, negative curvature results in rapid extinction via 'mutational meltdown', whereas positive curvature sometimes allows persistence. We speculate that fitness-dependent mutation rate may provide the conditions for genetic architecture to diverge between sexual and asexual taxa. PMID- 21635608 TI - Why migrate during the day: a comparative analysis of North American birds. AB - Migration can take place primarily during the day or at night, or during both depending on the species. Why the timing of migration varies among species has been the object of much research but the underlying ecological processes are still unclear. Proximally, migration timing may be influenced by the timing of other activities or may be more prevalent in species that migrate over long distances. Adaptive scenarios emphasize the reduction in flight costs at night especially in smaller species and the advantages of travelling in groups during the day to locate staging sites more efficiently. I used phylogenetic independent contrasts to examine these hypotheses in all North American nesting birds. I uncovered 24 evolutionary transitions in migration timing, most of which involved a switch from nocturnal to diurnal migration. Few of these transitions involved a concomitant change in the timing of foraging habits or migration distance. However, species in diurnal clades were larger, travelled in larger flocks and were generally more sociable than their nocturnal counterparts. The results support the hypotheses that a reduction in flight costs and the ability to pool information from companions are associated with migration timing in North American bird species. PMID- 21635609 TI - Electrocardiographic versus echocardiographic optimization of the interventricular pacing delay in patients undergoing cardiac resynchronization therapy. AB - INTRODUCTION: Echocardiographic optimization of the VV interval may improve CRT response, but it is time-consuming and not routinely performed. The aim of this study was to compare the response to cardiac resynchronization therapy (CRT) when the interventricular pacing (VV) interval was optimized by tissue Doppler imaging (TDI) to CRT response when it was optimized following QRS width criteria. METHODS AND RESULTS: The study included 156 consecutive CRT patients with severe heart failure and left bundle-branch block configuration. Atrioventricular interval was selected according to a pulsed Doppler assessment, and VV optimization was randomly assigned to echocardiography (ECHO group, n = 78) or electrocardiography (ECG group, n = 78). Optimal VV was defined for the ECHO group as producing the best LV intraventricular synchrony according to TDI displacement curves and for the ECG group as resulting in the narrowest QRS measured from the earliest deflection. At 6-month follow-up, percentage of echocardiographic responders (defined as neither death nor heart transplantation and a LV end-systolic volume reduction >10%) was higher in the ECG optimized group (50.0% vs 67.9%; P = 0.023), whereas clinical response (defined as neither death nor heart transplantation and >10% improvement in the 6-minute walking test) was similar in both groups (71.8% vs 73.1%; P = 0.858). CONCLUSIONS: VV optimization based on QRS width obtained a higher percentage of responders in terms of LV reverse remodeling compared to the TDI method. PMID- 21635610 TI - Electroanatomic properties of the pulmonary veins: slowed conduction, low voltage and altered refractoriness in AF patients. AB - INTRODUCTION: Rapid PV activity is critical in initiating and maintaining AF. The underlying substrate responsible for this remains uncertain. We sought to identify if patients with paroxysmal (PAF) and persistent atrial fibrillation (PeAF) have an abnormal substrate within the pulmonary veins (PVs). METHODS AND RESULTS: Thirty-nine patients with AF (21 PAF, 18 PeAF) were compared with 15 age matched controls with left-sided accessory pathways (AVRT). High-density 3D electroanatomic maps of the PVs were created. PV voltage, conduction, PV muscle sleeve length, effective refractory periods (ERPs) of the PVs, posterior left atrium (PLA), left atrial appendage (LAA) and distal coronary sinus (CSd), and signal complexity were assessed. Compared with controls, the PVs of AF patients had (1) lower mean-voltage and a higher % low-voltage; (2) shorter PV muscle sleeves; (3) slower conduction; (4) shorter ERP; and (5) more prevalent complex signals. Compared with the PAF group, the PeAF group had (1) higher % low voltage; (2) slower conduction; and (3) more complex signals. In PAF patients, the PLA and LAA ERPs were longer than controls and the PV ERP was shorter than controls; in PeAF patients PLA and LAA ERPs were reduced, but to a lesser extent than in the PVs. AF induction occurred during PV ERP testing in both AF groups, but not controls. CONCLUSIONS: PAF and PeAF patients demonstrate electrical and electroanatomic remodeling of the PVs compared to control patients without prior AF. Some of these changes were more marked in PeAF. PMID- 21635611 TI - The challenging face of focal atrial tachycardia in the post AF ablation era. AB - Radiofrequency ablation (RFA) for atrial fibrillation (AF) has become one of the most common catheter ablation procedures performed worldwide. As experience and success in treating patients with paroxysmal AF have increased, more centers are performing ablation for persistent AF. Optimal results may require ablation beyond the pulmonary veins with extensive biatrial substrate modification required in some cases to restore sinus rhythm. On the road to sinus rhythm atrial tachycardias are generally encountered either acutely within the index procedure or following. This has led to an increase in the frequency of focal atrial tachycardia and a need to review our understanding and approach to this and how it differs following substrate modification in contrast with the de novo setting. This review aims to describe the differences in responsible mechanism and its translation to mapping and ablation of focal AT particularly in the post ablation atria (paAT). PMID- 21635612 TI - beta-blockers protect against dispersion of repolarization during exercise in congenital long-QT syndrome type 1. AB - INTRODUCTION: beta-Blocker therapy reduces syncope and sudden death in long-QT syndrome type 1 (LQT1), but the mechanism of protection is incompletely understood. This study tested the hypothesis that beta-blockade reduces QT prolongation and dispersion of repolarization, measured as the T peak-to-end interval (T(pe) ), during exercise and recovery in LQT1 patients. METHODS AND RESULTS: QT and T(pe) were measured in 10 LQT1 patients (33 +/- 13 years) and 35 normal subjects (32 +/- 12 years) during exercise tests on and off beta-blockade. In LQT1 patients, beta-blockade reduced QT (391 +/- 25 milliseconds vs 375 +/- 26 milliseconds, P = 0.04 during exercise; 419 +/- 41 milliseconds vs 391 +/- 39 milliseconds, P = 0.02 during recovery) and markedly reduced T(pe) (91 +/- 26 milliseconds vs 67 +/- 19 milliseconds, P = 0.03 during exercise; 103 +/- 26 milliseconds vs 78 +/- 11 milliseconds, P = 0.02 during recovery). In contrast, in normal subjects, beta-blockade had no effect on QT (320 +/- 17 milliseconds vs 317 +/- 16 milliseconds, P = 0.29 during exercise; 317 +/- 13 milliseconds vs 315 +/- 14 milliseconds, P = 0.15 during recovery) and mildly reduced T(pe) (69 +/- 13 milliseconds vs 61 +/- 11 milliseconds, P = 0.01 during exercise; 77 +/- 19 milliseconds vs. 68 +/- 14 milliseconds, P < 0.001 during recovery). CONCLUSION: In LQT1 patients, beta-blockers reduced QT and T(pe) during exercise and recovery, supporting the theory that beta-blocker therapy protects LQT1 patients by reducing dispersion of repolarization during exercise and recovery. PMID- 21635613 TI - Between an ICD and a hard place. PMID- 21635614 TI - Cerebral magnetic resonance findings in asymptomatic patients after atrial fibrillation catheter ablation: cause for alarm or further study? PMID- 21635617 TI - Treatment of crow's feet while performing a rhytidectomy. PMID- 21635618 TI - The efficacy of autologous platelet rich plasma combined with ablative carbon dioxide fractional resurfacing for acne scars: a simultaneous split-face trial. AB - BACKGROUND: Ablative carbon dioxide (CO(2) ) fractional resurfacing is a promising therapeutic intervention for the treatment of acne scars, although this technique is associated with prolonged surgical site erythema and edema, which may affect the daily lives of patients. Autologous platelet-rich plasma (PRP) is known to enhance wound healing and has applications in many areas of medicine. OBJECTIVES: To evaluate the synergistic effects of autologous PRP with CO(2) fractional resurfacing for acne scars. MATERIALS AND METHODS: A split-face trial was conducted in 14 Korean participants with acne scars. All participants received one session of ablative CO(2) fractional resurfacing. Immediately after resurfacing, facial halves were randomly assigned to receive treatment with autologous PRP injections on one side (experimental side) and normal saline injections on the other side (control side). The participants were monitored for degree of recovery and resurfacing-associated adverse events, including prolonged erythema, edema, and other effects on days 0, 2, 4, 6, 8, 15, and 30. The intensity of erythema was objectively measured using a chromometer at the same time intervals. After one additional treatment session using the same protocol, two independent dermatologists evaluated clinical improvement using a quartile grading scale. RESULTS: All participants completed the study. Erythema on the experimental side improved faster than on the control side and was significantly less at day 4 (p=.01). This difference was confirmed using a chromometer (p=.049). Total duration of erythema was an average of 10.4+/-2.7 days on the control side and 8.6+/-2.0 days on the experimental side (p=.047). Edema also improved faster on the experimental side than on the control side. The total duration of edema was an average of 7.1+/-1.5 days on the control side and 6.1+/ 1.1 days on the experimental side (p=.04). Participants were also assessed for duration of post-treatment crusting, with a mean of 6.8+/-1.0 days on the control side and 5.9+/-1.1 days on the experimental side (p=.04). No other adverse effects were observed in any participant. Four months after the final treatment, overall degree of clinical improvement was significantly better on the experimental side (2.7+/-0.7) than on the control side (2.3+/-0.5) (p=.03). CONCLUSIONS: Treatment with PRP after ablative CO(2) fractional resurfacing enhances recovery of laser-damaged skin and synergistically improves the clinical appearance of acne scarring. PMID- 21635619 TI - 1,064-nm Q-switched neodymium-doped yttrium aluminum garnet laser and 1,550-nm fractionated erbium-doped fiber laser for the treatment of nevus of Ota in Fitzpatrick skin type IV. PMID- 21635621 TI - Hypertrophic scarring after burn scar treatment with a 10,600-nm carbon dioxide fractional laser. PMID- 21635622 TI - Sentinel node biopsy should be offered in thin melanoma with mitotic rate greater than one. AB - BACKGROUND: The American Joint Committee on Cancer recently added mitotic rate (MR) to their seventh edition staging system for melanomas, which went into effect on January 1, 2010. MR has replaced the Clark level of invasion for T1 melanomas. Dermatologists and dermatopathologists should be aware of these new guidelines. OBJECTIVE: To clarify how MR may be used to determine the prognosis of thin melanomas and to identify patients with thin melanomas who would be candidates for a sentinel lymph node biopsy (SLNB). METHODS: Reports in the literature were reviewed regarding mitotic rate in thin melanomas and the use of MR for prognosis and as an indication for SLNB. RESULTS: Multiple studies have shown MR to be a significant prognostic factor, surpassing Clark level of invasion, in patients with thin melanomas. SLNB is the best prognostic method for staging T1b (<1 mm thick with MR >=1 or ulceration), T2, T3, and T4 melanomas, and SLNB should be discussed and offered to these patients. CONCLUSIONS: Multiple studies support the use of MR in staging for thin melanomas, and patients with a high MR should be considered for SLNB. PMID- 21635627 TI - Treatment of acne scars in Asian patients using a 2,790-nm fractional yttrium scandium gallium garnet laser. AB - BACKGROUND: Treatment of atrophic scars using a fractional laser resurfacing technique has demonstrated favorable outcomes. OBJECTIVE: To evaluate the efficacy and safety of 2,790-nm-wavelength ablative fractional resurfacing on atrophic acne scars in Asian individuals. METHODS: Twenty participants (8 female, 12 male, mean age 26) with skin phototype IV and atrophic acne scars were treated with two sessions of 2,790-nm ablative fractional resurfacing laser at a 6-week interval. Objective and subjective (clinical evaluation by two blinded dermatologists) assessments were obtained at baseline and 1 and 3 months after the final treatment. RESULTS: At the 3-month follow-up, 70% of the participants were rated as having at least 50% to 89% improvement of scars. Mild erythema was the most common adverse effect, observed in 30% of participants, but resolved completely in an average of 5 days. CONCLUSIONS: Yttrium scandium gallium garnet ablative fractional resurfacing (2,790-nm) appears to be effective and well tolerated for the treatment of atrophic acne scars in Asians. The authors have indicated no significant interest with commercial supporters. PMID- 21635628 TI - Hematoxylin and eosin tissue stain in Mohs micrographic surgery: a review. AB - BACKGROUND: Several factors may affect Mohs micrographic surgery (MMS) tissue section quality. Although other factors may affect section integrity and ease of processing, tissue stains are the cornerstone of histologic diagnosis. When performed incorrectly, visualization and discrimination of microscopic details may be suboptimal and even impossible. Patient harm may result if an accurate diagnosis cannot be made. OBJECTIVE: To review hematoxylin and eosin (H&E) technique, the most commonly used staining technique in MMS tissue preparation. MATERIALS AND METHODS: Review of literature. Tissue sections were H&E stained using a linear stainer where one reagent container was omitted on each trial run to evaluate microscopic differences in staining quality. RESULTS: The chemical nature of H&E staining is discussed, along with commonly encountered problems, pitfalls, and troubleshooting tips. H&E stained sections are presented to demonstrate histologic appearance in the absence of certain reagents. CONCLUSIONS: The success of MMS depends on high-quality tissue sections. The staining process should be optimized to be reproducible and reliable. To readily identify and resolve poor staining quality, a firm understanding of the principles upon which tissue staining is based and its pitfalls is necessary. PMID- 21635629 TI - Tumor characteristics of Mohs surgery patients in Ottawa, Canada versus Houston, Texas--a consequence of access to care? AB - BACKGROUND: Ontario is one of the most underserved provinces in Canada for providing Mohs micrographic surgery (MMS). A new MMS clinic was opened in Ottawa, Ontario, in June 2009 to help combat the increasing incidence of nonmelanoma skin cancer (NMSC) in this region. OBJECTIVE: To prospectively compare MMS cases completed in Ottawa with cases completed in Houston, Texas, and examine the differences in tumor characteristics. MATERIALS AND METHODS: The first 150 cases performed in Ottawa were prospectively compared with 150 consecutive cases performed at a Mohs surgery clinic in Houston, Texas. Patient demographics, tumor diagnosis, primary or recurrent disease, tumor dimension, number of surgical stages, defect size, complexity of the procedure, and closure method were compared. RESULTS: The average preoperative tumor area was three times as great in Ottawa as in Houston. Almost one entire additional stage was required to clear the tumors treated in Ottawa. Postoperative defects were 87% larger in Ottawa. The number of advanced reconstructive repairs was significantly higher in Ottawa (93%) than Houston (14%). CONCLUSIONS: A significantly higher NMSC disease burden and greater surgical complexity was observed in the tumors treated in Ottawa than in Houston. PMID- 21635630 TI - A case of pincer nail treated using a shape-memory alloy. PMID- 21635631 TI - Subdermal minimal surgery with hyaluronic acid as an effective treatment for neck wrinkles. AB - BACKGROUND: Neck wrinkles are common, troublesome aesthetic deformities for which a limited number of treatments exist. Although previous study has demonstrated the safety and efficacy of subdermal minimal surgery technology in treatment of acne scarring, this technology has never been applied to treatment of wrinkles. OBJECTIVE: To evaluate the efficacy and safety of subdermal minimal surgery technology in the treatment of horizontal neck wrinkles. MATERIALS AND METHODS: All 12 enrolled participants were Koreans (Fitzpatrick skin types II-IV) with horizontal neck wrinkles. Participants underwent up to four sessions of treatment with subdermal minimal surgery technology at 4-week intervals. Wrinkle assessments were conducted at baseline and 2 months after the final treatment session. Participants and physicians evaluated improvement using pre- and post treatment photographs. RESULTS: All participants completed the study. Improvement of wrinkling by at least 50% was observed at 6 months (2 months after the final treatment session) in more than half of the participants, as determined according to physician and participant evaluation. CONCLUSION: Findings from this preliminary study demonstrate that use of subdermal minimal surgery technology results in an effective decrease of the appearance of neck wrinkles. PMID- 21635632 TI - Scalpel reduction of giant nasal folliculosebaceous hamartoma and reconstruction using porcine xenograft. PMID- 21635633 TI - Micropunch (0.8 mm or less in diameter) hair transplantation. PMID- 21635634 TI - Skin cancer arising in scars: a systematic review. AB - BACKGROUND: Despite numerous case reports, epidemiologic evidence regarding true rate of skin cancer in scars of any etiology is sparse. METHODS: Systematic literature review of all published epidemiologic studies on skin cancer in scar tissue from surgery, ulcers, or burns using citation databases and manual review. RESULTS: There were no epidemiologic data to quantify risk of skin cancer in surgical scars or chronic ulcers. Two eligible cohort studies were identified, from Denmark and Sweden, in which skin cancers in 16, 903 and 37,095 burn patients, respectively, were ascertained through cancer registry follow-up. Each reported standardized incidence ratios (SIRs) for skin cancer types on any site that were uniformly less than unity compared with the general population. Only the Danish cohort assessed skin cancers specifically on past burn injury sites and found a burn-site-specific SIR of 1.2 (95% confidence interval (CI)=0.4-2.7) for squamous cell carcinoma (SCC), 0.7 (95% CI=0.4-1.1) for basal cell carcinoma, and 0.3 (95% CI=0.0-1.2) for melanoma. CONCLUSIONS: Available epidemiologic data suggest that burn patients are not at higher risk of skin cancers in general, although a modest excess of SCC in burn scars cannot be excluded, nor can excess risk with longer follow-up. Risk of skin cancer in scars other than burn scars has not been investigated epidemiologically. PMID- 21635636 TI - Alimentary canal of the blow fly Chrysomya megacephala (F.) (Diptera: Calliphoridae): an emphasis on dissection and morphometry. AB - The alimentary canal is a major organ system that is often involved in the transmission of pathogens to humans from insects that serve as vectors of disease. In this study, we investigated the alimentary canal of the blow fly, Chrysomya megacephala (F.) (Diptera: Calliphoridae), highlighting the description for dissection and morphometric analysis of each organ. Dissection was performed in a phosphate buffer solution (pH=7.4) on 3(rd) instar larvae (three to four days old) and on both male and female adults (seven days old). Larval dissection was accomplished using two fine forceps to open the specimens from the posterior end and proceed anteriorly toward the cephalic segment. Meticulous dissection of the anterior end was vital for observation of the delicate salivary ducts, crop duct, and esophagus. Overall length of the 3(rd) instar alimentary canal measured 89.15 mm (range 81.40-99.70 mm). The midgut comprised the longest portion, measuring 46.35 mm (range 40.00-52.00 mm; n = 30) of the entire canal. Adult dissection was also performed from abdomen to head. Morphometric analyses revealed that the alimentary canal of males and females were relatively similar. No statistical differences were found between the entire length of the alimentary canal from mouth to anus (excluding all branches of the salivary glands, crop, and Malpighian tubules) of males and females. The alimentary canals of males measured 36.23 mm (range 32.60-41.20 mm) in length; whereas, those of females measured 37.23 mm (range 32.70-42.15 mm). Two-thirds of the entire canal length was comprised of midgut in each sex. PMID- 21635637 TI - Density of Ixodes scapularis ticks on Monhegan Island after complete deer removal: a question of avian importation? AB - Questing adult blacklegged tick (Ixodes scapularis Say) abundance declined markedly three years after the 1999 removal of white-tailed deer (Odocoileus virginianus Zimmermann) from Monhegan Island, ME. Since 2000, subadult ticks have not been found on Norway rats (Rattus norvegicus Berkenhout); questing nymphs have not been found since 2002. This suggested I. scapularis was reintroduced annually via bird importation of subadult ticks, but unable to complete its two year life cycle on the island due to lack of deer. To investigate this, we used uncertainty analysis to estimate 1) questing adult ticks/ha that would result from avian importation of nymphs, and 2) questing adult ticks/ha on Monhegan Island, using bird capture and tick burden data from Appledore Island, ME, flagged tick data from Monhegan Island, and ten uncertain parameters. During the deer-fed period (1990-2001), estimated tick density on Monhegan Island was 18 times greater than that of imported ticks. During the post-deer-fed period (2002 2008), Monhegan Island tick density was equivalent to imported tick density. This supported the premise that all I. scapularis ticks on Monhegan Island have been bird-derived since 2002. PMID- 21635638 TI - Strain diversity of Borrelia burgdorferi in ticks dispersed in North America by migratory birds. AB - The role of migratory birds in the dispersal of Ixodes scapularis ticks in the northeastern U.S. is well established and is presumed to be a major factor in the expansion of the geographic risk for Lyme disease. Population genetic studies of B. burgdorferi sensu stricto, the agent of Lyme disease in this region, consistently reveal the local presence of as many as 15 distinct strain types as designated by major groups of the ospC surface lipoprotein. Recent evidence suggests such strain diversity is adaptive to the diverse vertebrate hosts that maintain enzootic infection. How this strain diversity is established in emergent areas is unknown. To determine whether similar strain diversity is present in ticks imported by birds, we examined B. burgdorferi strains in I. scapularis ticks removed from migrants at an isolated island site. Tick mid-guts were cultured and isolates underwent DNA amplification with primers targeting ospC. Amplicons were separated by gel electrophoresis and sequenced. One hundred thirty seven nymphal ticks obtained from 68 birds resulted in 24 isolates of B. burgdorferi representing eight ospC major groups. Bird-derived ticks contain diverse strain types of B. burgdorferi, including strain types associated with invasive Lyme disease. Birds and the ticks that feed on them may introduce a diversity of strains of the agent of Lyme disease to emergent areas. PMID- 21635639 TI - Altitudinal variation in the morphometric characteristics of Aedes vexans Meigen from northeastern Turkey. AB - Body size is one of the most significant features of organisms and is correlated with a large number of ecological and physiological variables. Similar to size, biological shape is one of the most conspicuous aspects of an organism's phenotype and provides a link between the genotype and the environment. Body size may change with altitude and also habitat differences associated with altitude may affect the biological shape and some morphological characteristics. Four populations of Aedes vexans Meigen occurring in different ecological subregions at altitudes between 808-1,620 m in the Aras Valley from northeastern Turkey were compared using traditional and geometric morphometrics. When the wing shape differences of populations were analyzed by UPGM, the cluster analyses recognized two main groups of populations. Godekli (808 m) comprised the first group while Zulfikar (848 m), Surmeli (944 m), and Cilehane (1,620 m) populations comprised the second group. In the second group, Zulfikar and Cilehane populations showed a similar grouping pattern while Surmeli appeared as a different group. Centroid sizes were used as measures of overall wing size differences among different regions. Aedes vexans from the Surmeli region had relatively larger wings. PMID- 21635640 TI - Ecological surveillance of small mammals at Dagmar North Training Area, Gyeonggi Province, Republic of Korea, 2001-2005. AB - A seasonal rodent-borne disease surveillance program was established at Dagmar North Training Area located near the demilitarized zone, Republic of Korea, from 2001 through 2005. Selected habitats surveyed included earthen banks separating rice paddies, fighting positions along a 5 m rock-faced earthen berm, and extensive tall grasses with various degrees of herbaceous and scrub vegetation associated with dirt roads, rice paddies, ditches, ponds, or the Imjin River. Of the nine species of small mammals captured, the striped field mouse (Apodemus agrarius), the primary reservoir for Hantaan virus, was the most frequently collected, representing 92.5% of the 1,848 small mammals captured. Males were captured similarly to females during the spring and summer seasons but were captured less frequently during the fall and winter seasons. Gravid rates were highest in the fall (25.5-57.3%) with the lowest rates during the summer (0.0 2.2%). Capture rates were the lowest along earthen banks separating rice paddies (5.5%) and highest in unmanaged tall grasses and crawling vegetation (15.3 43.5%). An increased knowledge of ecological factors that impact the abundance and distribution of small mammals and the associated ectoparasites and pathogens they harbor is critical for developing accurate disease risk assessments and mitigation strategies for preventing vector- and rodent-borne diseases among soldiers training in field environments. PMID- 21635641 TI - Evaluation of ammonia, human sweat, and bovine blood as attractants for the female canyon fly, Fannia conspicua (Diptera: Muscidae), in southern California. AB - Human sweat, liquid ammonia, and bovine blood are known to be attractive to some hematophagous flies. These materials were evaluated with and without carbon dioxide (CO(2)) for their ability to increase capture of female "canyon flies" (Fannia conspicua Malloch) using CDC-type suction traps (without light). Ammonia acted synergistically with CO(2) to increase trap catch 89.9% over CO(2) alone. There was no synergistic effect of human sweat or bovine blood with CO(2). In the absence of CO(2), none of the three materials increased trap catch of female canyon flies relative to non-baited traps. Implications for canyon fly control and further trap improvement are discussed. PMID- 21635642 TI - Susceptibility of adult mosquitoes to insecticides in aqueous sucrose baits. AB - Mosquitoes characteristically feed on plant-derived carbohydrates and honeydew just after emergence and intermittently during their lives. Development of toxic baits focusing on this carbohydrate-seeking behavior may potentially contribute to localized control. In the present study, ten insecticides were fed to female Culex quinquefasciatus, Anopheles quadrimaculatus, and Aedes taeniorhynchus in a 10% sucrose solution. Active ingredients representative of five classes of insecticides (pyrethroids, phenylpyroles, pyrroles, neonicotinoids, and macrocyclic lactones) were selected for comparison with commercial formulations used to facilitate incorporation of active ingredients into aqueous sucrose solutions. Sucrose as a phagostimulant significantly enhanced mortality to toxicants. In general, the most effective active ingredients were fipronil, deltamethrin and imidacloprid, followed by spinosad, thiamethoxam, bifenthrin, permethrin, and cyfluthrin. The least effective ingredients were chlorfenapyr and ivermectin. For some of the ingredients tested, Cx. quinquefasciatus was the least susceptible species. One-day-old male Cx. quinquefasciatus were more susceptible than females; however, no differences existed between one- and seven day-old mosquitoes. There were no differences in susceptibility between unfed and gravid ten-day-old female Cx. quinquefasciatus to bifenthrin. In conclusion, several pesticides from different classes of compounds have potential for use in development of toxic baits for mosquitoes. PMID- 21635643 TI - Comparison of the efficacy of CO2-baited and unbaited light traps, gravid traps, backpack aspirators, and sweep net collections for sampling mosquitoes infected with Japanese encephalitis virus. AB - Two field studies were conducted to determine the efficacy of mosquito collection methods for species composition, species abundance, and Japanese encephalitis virus infection rates in Taiwan. Traps evaluated included John W. Hock (JH) model UD black light traps, JH model 1012 new standard miniature CDC light traps, JH model 1712 CDC gravid traps, and Taiwan-made Pest-O-Lite light traps. Backpack aspirators and sweep nets were also used to collect the resting population. Culex tritaeniorhynchus in all studies and Mansonia uniformis in the Taipei areas were the two most abundance species collected. Dry ice-baited UD black light traps were effective in regard to species diversity, species abundance, and Japanese encephalitis virus infection rates. The unbaited Pest-O-Lite light traps collected significantly more female mosquitoes than the UD black light traps but performed similarly with regard to species diversity and male mosquito collection. Most mosquitoes collected by Pest-O-Lite light traps were dried and not suitable for virus detection. Dry ice-baited CDC light traps collected significantly fewer mosquitoes than other light traps. Although CO(2) -baited UD black light traps with octenol attracted more mosquitoes, no statistical significance was found compared to CO(2) -baited UD black light traps without octenol. Japanese encephalitis viruses were isolated from half of the positive pools in UD black light traps and CDC light traps. PMID- 21635644 TI - Abundance of West Nile virus mosquito vectors in relation to climate and landscape variables. AB - It is currently unclear if the potential for West Nile virus transmission by mosquito vectors in the eastern United States is related to landscape or climate factors or both. We compared abundance of vector species between urban and suburban neighborhoods of Henrico County, VA, in relation to the following factors: temperature, precipitation, canopy cover, building footprint, and proximity to drainage infrastructure. Mosquitoes were collected throughout the 2005, 2006, and 2007 seasons and tested for West Nile virus (WNV) in pools of 10 50. Test results of mosquito pools were compared to average site abundance from 37 sites in Henrico County, VA; abundance was then examined in relation to ecological variables. Urban infrastructure was positively correlated with the abundance of Culex pipiens L./Cx. restuans, and our findings implicate combined sewer overflow systems as large contributors to Culex vector populations. No measure of urbanization examined in our study was correlated with Aedes albopictus abundance. Our study showed that certain landscape variables identified using Geographic Information Systems are valuable for predicting primary WNV vector abundance in Virginia, and that temperature along with low precipitation are strong predictors of population growth. Our results support other regional studies that found WNV proliferates under drought conditions. PMID- 21635645 TI - Generation of monoclonal antibodies to vertebrate albumins for analysis of arthropod blood meals. AB - An immunoassay using monoclonal antibodies (MAbs) that are specific for different vertebrate taxa (from class to species) has been developed that simplifies and facilitates analysis of vertebrate blood meals from arthropod vectors. The MAbs have been prepared against the single protein albumin, the most abundant protein in vertebrate sera. A panel of these antibodies has been generated against albumins from 33 species of vertebrates, representing four classes, 15 orders, and 25 families. Immunoreactivity of albumin in mosquito blood meals can be detected as late as 48 h after feeding. Immunoassays with MAbs can be carried out in the field as well as the laboratory. Used in conjunction with nucleic acid assays or used alone with an appropriate assortment of antibodies, the assay is simple, sensitive, and unambiguous. PMID- 21635646 TI - Cold season mortality and hatching behavior of Aedes aegypti L. (Diptera: Culicidae) eggs in Buenos Aires City, Argentina. AB - In temperate regions, populations of Aedes aegypti survive the cold season in the egg stage. In the present work, we studied the cold-season mortality of Ae. aegypti eggs and their subsequent hatching pattern in Buenos Aires city. Eggs were exposed during the winter season (three months) in three neighborhoods located along a gradient of distance toward the Rio de la Plata River, coincident with a gradient of activity of Ae. aegypti. Results showed mortalities lower (30.6%) than those from tropical regions during the dry season. Significant differences were detected among the egg mortalities of each site with a maximum value at the site nearest the Rio de la Plata River (50%), and a minimum value at the most continental site (9%). Post-experimental hatching response of eggs differed between sites, with the highest proportion of hatched eggs during the first immersion in the site nearest to the river and the lowest proportion in the most continental site. The hatching proportion also differed between age classes, with older (early-laid) eggs hatching later than new (late-laid) ones. Our results provide the first information of Ae. aegypti egg mortality in temperate South America and support the hypothesis that differences in egg mortality are associated with abundance patterns of Ae. aegypti in Buenos Aires city. PMID- 21635647 TI - Field evaluation of imidacloprid as a systemic approach to flea control in black tailed prairie dogs, Cynomys ludovicianus. AB - Epizootic outbreaks of sylvatic plague have dramatically influenced prairie dog (Cynomys sp.) populations across North America. While a great deal of debate surrounds the cause and persistence of plague, flea control can stop the spread of plague epizootic outbreaks and even increase prairie dog survival under non epizootic conditions. We investigated a newly-developed imidacloprid-treated grain bait that could potentially reduce flea infestations and mitigate the effects of plague on black-tailed prairie dogs (C. ludovicianus). We used a study design involving randomly assigned experimental and control study plots to assess the effectiveness of the systemic flea control product. We observed a significant difference in flea prevalence and abundance between experimental and control sites on three of the four sites treated with a single application of imidacloprid-treated grain bait for up to 90 days post-treatment. We observed an even greater reduction in flea infestations following the double application of treatment bait on two of three additional experimental sites. While we were unable to reduce flea infestations to the extent reported for more commonly used topical insecticides containing deltamethrin, imidacloprid might still be effective at reducing the risk of plague and halting epizootics. In addition, this systemic product can be more rapidly applied than topical insecticides, providing managers with a tool to quickly reduce flea infestations. Future research is needed to evaluate the effectiveness of different application timing and rates, the utility of the product in limiting plague, and the potential effects on non-target species that might also consume the treated bait. PMID- 21635648 TI - Surveillance of the chikungunya vector Aedes albopictus (Skuse) in Emilia-Romagna (northern Italy): organizational and technical aspects of a large scale monitoring system. AB - The chikungunya virus outbreak that occurred in 2007 in northern Italy (Emilia Romagna region) prompted the development of a large scale monitoring system of the population density of Aedes albopictus (Skuse, 1894), comparable at the provincial and municipal levels. In 2007, egg density data presented an aggregated distribution (VMR >1) and Taylor's power law was applied to calculate the minimum number of ovitraps needed to obtain the prefixed precision levels: D=0.2 in the areas where the chikungunya epidemic occurred and D=0.3 in all the other urban areas >600 ha. The estimated minimum ovitrap number was then used to set up a monitoring network at the regional scale in season 2008 (May-October). In 242 municipalities 2,741 ovitraps were activated and the 2008 sampled data showed a similar aggregated distribution as in 2007. The adequacy of the monitoring design was evaluated by recalculating the Taylor's coefficients and the minimum ovitrap number for each urban area >600 ha using the 2008 egg density data. The comparison between the two estimates showed that the minimum ovitrap number calculated in 2007 was underestimated by 2.7% in weeks 22-41 but was overestimated by 29.4% if referring to the period of highest population density (weeks 27-37). The low cost of the proposed monitoring system, based on the use of fortnightly checked ovitraps, could make it economically sustainable even in a non-epidemic season. PMID- 21635649 TI - Seasonal and among-site variation in the occurrence and abundance of fleas on California ground squirrels (Otospermophilus beecheyi). AB - An improved understanding of the ecology of fleas on California ground squirrels, Otospermophilus beecheyi, is warranted given the role of fleas in the transmission, and perhaps persistence, of the plague-causing bacterium Yersinia pestis. We sampled O. beecheyi on a seasonal basis from three study sites, each representing a different land use type (preserve, pasture, and agriculture) in the San Joaquin Valley, CA. Overall, the abundance of fleas on squirrels was greatest in spring at the preserve site, in summer at the agriculture and pasture sites, and in winter at the pasture site. Hoplopsyllus anomalus, the species most frequently found on squirrels, was most abundant in spring at the preserve site and in summer at the agriculture and pasture sites. Oropsylla montana was most abundant in winter at the pasture site and on adult squirrels. Echidnophaga gallinacea was most abundant in fall on juvenile squirrels at the preserve site. All three flea species we encountered are known to be potential vectors of Y. pestis. Future efforts to predict flea species occurrence and abundance (and plague risk) at sites of concern should consider seasonal microclimatic conditions and the potential influence of human land use practices. PMID- 21635650 TI - Distribution and larval habitats of Anopheles species in northern Gyeonggi Province, Republic of Korea. AB - A total of 180 larval collection sites (e.g., rice paddies, marshes, ground pools, ponds, stream margins, and irrigation and drainage ditches) was surveyed within a 2 km radius from Warrior Base training area, 5 km south of Panmunjeom (Joint Security Area, demilitarized zone), Gyeonggi Province, Republic of Korea (ROK), from May through October, 2007 to characterize larval habitat distributions of members of the Anopheles Hyrcanus Group (An. sinensis, An. lesteri, An. pullus, An. belenrae, An. kleini, and An. sineroides). A total of 5,859 anopheline larvae was collected from 84.4% of the sites surveyed, of which 4,071 were identified to species by polymerase chain reaction (PCR) using the ribosomal DNA internal transcribed spacer 2 (rDNA ITS2). Anopheles sinensis (52.6%) was the most frequently collected, followed by An. kleini (29.4%), An. sineroides (9.8%), An. pullus (6.7%), An. belenrae (1.1%), and An. lesteri (0.5%). Anopheles pullus and An. kleini were collected in greater proportions in May and from May - July, respectively. Few An. sinensis were collected from May - June, but it was the predominant species collected by August, and accounted for >80% of all larvae from September - October. Anopheles kleini was found in all habitats sampled; however, it was collected most frequently in young growth rice paddies, while An. sinensis was collected more frequently in mature and post harvest paddies. Anopheles pullus was associated with pre-cultivated rice paddies, including water-filled tire ruts left from the previous fall's harvest. PMID- 21635651 TI - Meteorological indicators for Amblyomma cajennense and population dynamics in the Tamaulipan Biotic Province in Texas. AB - We studied the population dynamics of free-living ticks in the Tamaulipan Biotic Province in south Texas from March, 2005 to November, 2008. We collected 70,873 ticks using carbon dioxide traps. Amblyomma cajennense represented 93.6% of the ticks identified. A. cajennense is distributed from northern Argentina to south Texas in the Tamaulipan Biotic Province. Emergence of larval A. cajennense ticks was observed two to five weeks after significant rain events (p<0.0001) and had a strong negative correlation with temperature (p<0.0001). More larvae were observed under humid conditions (p<0.05). Fewer larvae were observed during windy and warmer conditions (p<0.05). This observation indicates high sensitivity of larvae to desiccating conditions. Peaks in nymphal activity were observed after peaks of larval emergence. Activity of nymphs was negatively correlated with temperature (p<0.05). Adult activity was negatively correlated with humidity (p<0.05) and negatively correlated with total rain from three to six weeks prior to observation (p<0.05). Adult A. cajennense are particularly tolerant to drier conditions relative to other closely related ticks. Adult female activity was positively correlated with temperature (p<0.05). Peaks in rain activity and a summer behavioral diapause appear to be the dominant factors controlling emergence of larvae, and by extension, the life cycle of A. cajennense in the Tamaulipan Biotic Province. PMID- 21635652 TI - Prospective field study of transovarial dengue-virus transmission by two different forms of Aedes aegypti in an urban area of Bangkok, Thailand. AB - A prospective field study was conducted to determine transovarial dengue-virus transmission in two forms of Aedes aegypti mosquitoes in an urban district of Bangkok, Thailand. Immature Aedes mosquitoes were collected monthly for one year and reared continuously until adulthood in the laboratory. Mosquitoes assayed for dengue virus were processed in pools and their dengue virus infection status was determined by one-step RT-PCR and nested-PCR methods. Of a total 15,457 newly emerged adult Ae. aegypti, 98.2% were dark and 1.8% of the pale form. The results showed that the minimum infection rate (MIR) by transovarial transmission (TOT) of dengue virus during the one-year study ranged between 0 to 24.4/1,000 mosquitoes. Dengue virus TOT increased gradually during the hot summer months, reaching a peak in April-June, while dengue cases peaked in September, a rainy month near the end of the rainy season. Therefore, mosquito infections due to TOT were prevalent four months before a high incidence of human infections. TOT dengue virus infections occurred in both forms of Ae. aegypti. All four dengue serotypes were detected, with DEN-4 predominant, followed by DEN-3, DEN-1, and DEN-2, respectively. PMID- 21635653 TI - Morphometric and morphological variation between two different populations of Phlebotomus major s.l. from endemic and non-endemic foci of visceral leishmaniasis in Iran. AB - Populations of Phlebotomus major were examined in two endemic and nonendemic foci of visceral leishmaniasis in Iran. Based on the shape of the aedeagus and ventrally located hairs of coxite and pharyngeal armatures, two morphotypes were found sympatrically in the endemic area of Borazjan. Significant differences in morphometric survey were observed in at least 11 measured characters. The aedeagus of the non-endemic Miyandoab morphotype, and also of a few specimens from Borazjan, is completely parallel throughout its length with a slightly expanded end. Ventrally located hairs of the middle coxite were longer and more compact. It is close morphologically to P. major neglectus (P. neglectus), which was recently recorded from Iran. It is also morphologically similar to P. notus, which has not yet been reported from Iran and needs further investigation. The aedeagus of the morphotype occurring only in Borazjan is narrower in the middle and the hairs are closer to the base of the coxite and are shorter and more outspread, which makes it similar to P. major krimensis or P. neglectus. The two morphotypes occurring sympatrically in Borazjan do not appear to be subspecies and it may be premature to propose them as separate species. Further investigation is needed to clarify the actual status of P. major s. l. in Iran. PMID- 21635654 TI - Temporal and spatial distribution of malaria within an agricultural settlement of the Brazilian Amazon. AB - Malaria has reemerged in tropical regions with rapid population growth and deforestation. The dynamics of malaria transmission in agricultural settlements of the Amazon have been poorly defined. We studied the spatial distribution of malaria incidence in Roraima, Brazil, using multi regression analysis on 12 parameters that described social, housing, and behavioral variables. Malaria cases were associated with the proximity of Anopheles darlingi breeding sites, the main vector in these areas. During the dry season, transmission was enhanced near a temporary river. Cases occurred throughout the year near fish-farming dams. Epidemiological models derived from urban or riverine malaria are probably inadequate for describing disease transmission in agricultural settlements, where cases are clustered near breeding sites, while the majority of the population remains unaffected. Identification of these areas, associated with residual insecticide spraying or surveillance, may considerably decrease the costs of control efforts. PMID- 21635655 TI - Heteropteran insects as mosquito predators in water jars in southern Vietnam. AB - Residents of Vietnam living in areas with water shortages and/or poor tap water maintain water storage containers, such as jars, in and around their domiciles in order to store water used in daily life. Although these water jars are known to be important breeding sources of the Aedes mosquito, use of chemical larvicides in such containers is legally prohibited in Vietnam. In this study, we identified the dominant mosquito insect predators in water jars in and around residences located in Tan Chanh, Long An, southern Vietnam. Of 3,646 Heteroptera collected from such jars, Corixidae (Micronecta spp.) and Veliidae (Microvelia spp.) were revealed to be the dominant predators. Polymerase chain reaction (PCR) analysis revealed that 40% of Micronecta and 12% of Veliidae had Aedes aegypti-positive reactions, indicating that these two dominant Heteroptera are important predators of Ae. aegypti. Our results suggest that aquatic Heteroptera may be an important mosquito control agent in addition to the currently used copepods. PMID- 21635656 TI - Ecological aspects of mosquitoes (Diptera: Culicidae) in an Atlantic forest area on the north coast of Rio Grande do Sul State, Brazil. AB - Mosquito diversity was determined in an area located on the southern limit of the Atlantic Forest on the north coast of Rio Grande of Sul State. Our major objective was to verify the composition, diversity, and temporal distribution of the mosquito fauna, and the influence of temperature and rainfall. Samplings were performed monthly between December, 2006 and December, 2008, in three biotopes: forest, urban area, and transition area, using CDC light traps and a Nasci vacuum. A total of 2,376 specimens was collected, from which 1,766 (74.32%) were identified as 55 different species belonging to ten genera. Culex lygrus, Aedes serratus, and Aedes nubilus were dominant (eudominant) and constant throughout samplings. The forest environment presented the highest species dominance (D(S) =0.20), while the transition area showed the highest values of diversity (H'=2.55) and evenness (J'=0.85). These two environments were the most similar, according to the Morisita-Horn Index (I(M-H) =0.35). Bootstrap estimates showed that 87.3% of the species occurring in the region were detected. The seasonal pattern showed a greater abundance of mosquitoes between May and October, indicating the period to intensify entomological surveillance in that area. PMID- 21635657 TI - Seasonal occurrence of black flies (Diptera: Simuliidae) in a desert stream receiving trout farm effluent. AB - The distribution and abundance of black flies (Diptera: Simuliidae) in a small desert stream were influenced by environmental changes caused by recharge of water supply storage basins and an aquaculture operation. Simulium virgatum was the most abundant benthic insect collected in Whitewater Canyon (Riverside County, CA) after April; however, it was never found in trout farm effluent where the ammonium-nitrogen concentration was > 0.25 mg/liter. S. virgatum densities downstream of the input of water from the Colorado River aqueduct were lower than at other sampling sites in the Whitewater River. Simulium tescorum, an especially anthropophilic black fly, was most abundant during February and March, was not collected from late spring through early autumn, and was found only in the highly enriched, less variable flow of trout farm effluent. The mean concentrations of ammonium nitrogen and nitrate nitrogen in the trout farm effluent nearly 1 km from the fish holding ponds were ten and two times, respectively, the ambient levels in the Whitewater River upstream of the effluent discharge point. A combination of factors probably contributed to the presence of S. tescorum in the trout farm effluent including homogenization of the flow regime, enrichment of larval resources, and the development of riparian vegetation that provided oviposition and attachment sites. PMID- 21635658 TI - The importance of an invasive tree fruit as a resource for mosquito larvae. AB - Invasive plants are common and may provide resources through litter for container mosquito larvae. Invasive plant reproductive parts can make up a substantial part of litter but have mostly been ignored as a resource for mosquito larvae. We hypothesized that the reproductive fruits of the invasive eastern red cedar, Juniperus virginiana, provide high quality resources for the invasive, container mosquito Aedes albopictus at the western margin of its invasive range in North America. To test this hypothesis, we performed two laboratory experiments. The first examined the response of individual larvae of Ae. albopictus to different amounts of J. virginiana leaf (fresh and senesced) and J. virginiana fruit (ripe and unripe), as well as to a control leaf (Quercus virginiana, live oak). The second experiment examined the response of different densities of Ae. albopictus larvae to each litter type. We found significant differences in response by individual larvae to different amounts of litter and litter types. We also found J. virginiana litter components could support positive population growth rates as a function of initial larval density where the control leaf could not. We conclude that invasive plants may provide high quality resources, and that the reproductive parts (fruits, flowers, cones) may be an important and overlooked component in provisioning larval habitats. Therefore, the expansion of J. virginiana into grassland areas may contribute to the expansion of Ae. albopictus westward in North America. PMID- 21635659 TI - Frequency of pyrethroid resistance in Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in Thailand. AB - Thirty-two Aedes aegypti populations collected throughout Thailand and five populations of Aedes albopictus from southern Thailand were subjected to standard WHO contact bioassays to assess susceptibility to three commonly used synthetic pyrethroids: permethrin, deltamethrin, and lambda-cyhalothrin. A wide degree of physiological response to permethrin was detected in Ae. aegypti, ranging from 56.5% survival (Lampang, northern Thailand) to only 4% (Kalasin in northeastern and Phuket in southern Thailand). All 32 populations of Ae. aegypti were found to have evidence of incipient resistance (62.5%) or levels of survival deemed resistant (37.5%) to permethrin. Four populations of Ae. albopictus were found with incipient resistance (97 - 80% mortality) and one with resistance (< 80%) to permethrin. The majority of Ae. aegypti populations (68.7%) was susceptible (> 98% mortality) to deltamethrin, with incipient resistance (observed 97-82% mortality) in other localities. In contrast, all populations of Ae. aegypti were completely susceptible (100% mortality) to the recommended operational dosage of lambda-cyhalothrin. All five populations of Ae. albopictus were found completely susceptible to both deltamethrin and lambda-cyhalothrin. Evidence of defined incipient or resistance to synthetic pyrethroids mandates appropriate response and countermeasures to mitigate further development and spread of resistance. In light of these findings, we conclude that routine and comprehensive susceptibility monitoring of dengue mosquito vectors to synthetic pyrethroids should be a required component of resistance management policies and disease control activities. PMID- 21635660 TI - Implications of Plasmodium parasite infected mosquitoes on an insular avifauna: the case of Socorro Island, Mexico. AB - Avian malaria (Plasmodium spp.) has been implicated in the decline of avian populations in the Hawaiian Islands and it is generally agreed that geographically isolated and immunologically naive bird populations are particularly vulnerable to the pathogenic effects of invasive malaria parasites. In order to assess the potential disease risk of malaria to the avifauna of Socorro Island, Mexico, we surveyed for Plasmodium isolates from 1,300 resident field-caught mosquitoes. Most of them were identified as Aedes (Ochlerotatus) taeniorhynchus (Wiedemann, 1821), which were abundant in the salt marshes. We also collected Culex quinquefasciatus Say, 1823 close to human dwellings. Mitochondrial ND5 and COII gene sequences of Ae. taeniorhynchus were analyzed and compared to corresponding sequences of mosquitoes of the Galapagos Islands, Latin America, and the North American mainland. Aedes lineages from Socorro Island clustered most closely with a lineage from the continental U.S. Plasmodium spp. DNA was isolated from both species of mosquitoes. From 38 positive pools, we isolated 11 distinct mitochondrial Cytb lineages of Plasmodium spp. Seven of the Plasmodium lineages represent previously documented avian infective strains while four were new lineages. Our results confirm a potential risk for the spread of avian malaria and underscore the need to monitor both the mosquito and avian populations as a necessary conservation measure to protect endangered bird species on Socorro Island. PMID- 21635661 TI - Serological study of the 2009 pandemic due to influenza A H1N1 in the metropolitan French population. AB - We looked for evidence of antibodies to the 2009 influenza A/H1N1 pandemic virus in panels of sera from individuals living in metropolitan France, obtained either before, during or after the epidemic, using standard haemagglutination inhibition and microneutralization tests. The difference between seroprevalence values measured in post- and pre-epidemic panels was used as an estimate of seroconversion rate in different age groups (23.4% (0-24 years, age-group 0); 16.5% (25-34); 7.9% (35-44); 7.2% (45-54); 1.6% (55-64); and 3.1% (>65)), confirming that the distribution of cases in different age groups was similar to that of the seasonal H1N1 virus. During the pre-pandemic period low-titre cross reactive antibodies were present in a large proportion of the population (presumably acquired against seasonal H1N1) whereas cross-reactive antibodies were detected in individuals over the age of 65 years with significantly higher prevalence and serological titres (presumably acquired previously against Spanish flu-related H1N1 strains). Clinical data and analysis of post-pandemic seroprevalence showed that few of these latter patients were infected by the influenza virus during the epidemic. In contrast, the majority of both clinical cases and seroconversions were recorded in the 0-24 age group and a global inverse relationship between prevalence of antibodies to pH1N1 in the pre pandemic period and rate of seroconversion was observed amongst age groups. Our results emphasize the complex relationships involved in antigenic reactivity to pandemic and seasonal H1N1 viral antigens; hence the difficulty in distinguishing between low-titre specific and cross-reactive antibodies, establishing precise seroprevalence numbers and fully understanding the relationship between previous immunity to seasonal viruses and protection against the novel variant. PMID- 21635662 TI - Susceptibility of European Escherichia coli clinical isolates from intra abdominal infections, extended-spectrum beta-lactamase occurrence, resistance distribution, and molecular characterization of ertapenem-resistant isolates (SMART 2008-2009). AB - A total of 3160 clinical isolates of Escherichia coli from intra-abdominal infections were collected during 2008-2009 from 13 European countries. The frequency of extended-spectrum beta-lactamase (ESBL)-producing isolates in Europe was 11%. The most active antibiotics tested were typically imipenem, ertapenem, and amikacin, although the activity of all non-carbapenem antibiotics was lower when tested against ESBL-positive isolates than when tested against ESBL-negative isolates. Ertapenem exhibited 99.3% susceptibility with all isolates, and 96.8% susceptibility with ESBL-positive isolates. With application of the ertapenem CLSI clinical breakpoint for resistance (MIC >=1 mg/L), only six isolates (0.2%) were ertapenem-resistant, and only three of these were available for molecular characterization. Of those three, only one was ESBL-positive (CTX-M-14), and two were carbapenemase-positive (OXA-48). All three were negative for, VIM, NDM and KPC carbapenemases. Although the level of ertapenem resistance in E. coli is very low, further monitoring of ertapenem susceptibility and molecular characterization of ertapenem-resistant isolates is needed. PMID- 21635663 TI - Carbapenemase-producing Klebsiella pneumoniae: (when) might we still consider treating with carbapenems? AB - Infections caused by carbapenemase-producing Klebsiella pneumoniae (CPKP) are increasing in frequency worldwide. CPKP isolates exhibit extensive drug resistance phenotypes, complicate therapy, and limit treatment options. Although CPKP isolates are often highly resistant to carbapenems, a proportion of these have relatively low MICs for carbapenems, raising the question of whether this class of agents has any therapeutic potential against CPKP infections. Results from animal studies and patient outcome data indicate that carbapenems retain meaningful in vitro activity against CPKP isolates with carbapenem MICs of <= 4 mg/L. Accumulating clinical experience also suggests that the therapeutic efficacy of carbapenems against CPKP isolates with MICs of <= 4 mg/L is enhanced when these agents are administered in combination with another active antibiotic. The results of human pharmacokinetic/pharmacodynamic studies are in line with the above observations; it is highly probable that a high-dose/prolonged-infusion regimen of a carbapenem would attain a time above the MIC value of 50% for CPKP isolates with MICs up to 4 mg/L, ensuring acceptable drug exposure and favourable treatment outcome. The analyses summarized in this review support the notion that carbapenems have their place in the treatment of CPKP infections and that the currently proposed EUCAST clinical breakpoints could direct physicians in making treatment decisions. PMID- 21635664 TI - Update on emergence of HIV-1 resistance to antiretroviral drug classes in an Italian national database: 2007-2009. AB - We analysed trends of human immunodeficiency virus type 1 (HIV-1) drug resistance during 2007-2009 in the Italian national HIV drug resistance database 'ARCA'. Prevalence of resistance in each year was examined on the basis of the presence of major International AIDS Society-2009 mutations. Predictors of resistance were analysed by multivariable logistic regression. Nine hundred and sixty-six patients were selected. Resistance to nucleoside reverse transcriptase inhibitors and protease inhibitors showed a significant decline with respect to previous surveys. Resistance to any class of drug and three drug classes remained stable. Independent predictors of three-class resistance were the number of treatment regimens experienced, prior suboptimal nucleoside reverse transcriptase inhibitor therapy and the current use of ritonavir-boosted protease inhibitors. PMID- 21635665 TI - Triple flap vaginoplasty for agenesis of the vagina. AB - AIM: Various procedures have been described for the creation of neovagina. McIndoe operation is gold standard for vaginoplasty in which split thick skin graft is used for lining the neovagina. However the rate of contraction of the graft is very high. We have used labia minora flaps along with the third flap created from suburethral tissue for lining the neovaginal space in 17 patients with good results. METHODS: The method was used in 17 patients. After creation of neovaginal space three flaps were raised - two labia minora flaps and the third flap from suburethral tissue. The flaps were inturned and lined the neovaginal space. If whole space was not lined with flaps, supplementary grafting was used to cover the unlined space. RESULTS: Thirteen patients came for follow up out of whom in one patient the labia minora flap necrosed and patient developed contracture of the vagina. In all other patients the result was satisfactory. There was no need of added lubrication during sex in any of the patients. CONCLUSION: A three flap method of vaginoplasty in agenesis of the vagina has been described with good result. PMID- 21635667 TI - Polycyclic aromatic hydrocarbons induce migration in human hepatocellular carcinoma cells (HepG2) through reactive oxygen species-mediated p38 MAPK signal transduction. AB - Although polycyclic aromatic hydrocarbons (PAHs) are carcinogenic and have been extensively studied with regard to tumor formation, few studies have investigated the involvement of these environmental chemicals in tumor migration and invasion. Polycyclic aromatic hydrocarbons induce reactive oxygen species (ROS) and activate MAPK signal transduction. The p38 signaling transduction pathway, one of the most typical MAPK pathways, plays an essential role in regulating cell migration. Therefore, we investigated whether three PAHs, benzo[a]anthracene (B[a]A), benzo[k]fluoranthene (B[k]F), and indeno[1,2,3-c,d]pyrene (IND), induce migration in human hepatocellular carcinoma cell line HepG2 through ROS-mediated p38 MAPK signal transduction. Reactive oxygen species generation and p38 MAPK activity both increased in a dose-dependent manner and were prevented by SB203580, an inhibitor of p38 MAPK, and N-acetylcysteine (NAC), a ROS scavenger. Expression of migration-related genes was also increased by B[a]A, B[k]F, and IND in a dose-dependent manner and was inhibited by SB203580 and NAC. The migration of HepG2 cells, observed using the Transwell migration assay, also increased in a dose-dependent manner and was prevented by SB203580 and NAC. Our results indicate that the ROS-mediated p38 MAPK signaling pathway plays an essential role in the PAH-induced migration of HepG2 cells. PMID- 21635668 TI - Unstimulated whole saliva creatine phosphokinase in acute myocardial infarction. AB - OBJECTIVE: Accurate and rapid diagnosis of acute myocardial infarction (MI) is of major medical and economic importance. The objective of this study was to identify unstimulated whole saliva creatine phosphokinase (CPK) in patients with acute MI. SUBJECTS AND METHODS: A case-control study was carried out in 30 normal healthy individuals and 30 patients with acute MI were hospitalized in CCU of Kamkar hospital, Qom, Iran. CPK levels were assayed in serum and unstimulated whole saliva at the first and the second days of acute MI by IFCC method. Statistical analysis of the Student's t-test and Pearson correlation coefficient was performed. RESULTS: The mean saliva and serum levels at both the first and the second days of acute MI were significantly higher in patients with acute MI compared with healthy individuals. They were significantly greater in the first day than in the second day. Saliva CPK concentration correlated significantly with serum CPK level in the first day (r = 0.442, P < 0.01) and in the second day of acute MI (r = 0.268, P < 0.01). CONCLUSION: Results suggest that salivary CPK can be used as an alternative to serum CPK for diagnosis and monitoring of myocardial infarction. PMID- 21635669 TI - Ethanol consumption and pineal melatonin daily profile in rats. AB - It is well known that melatonin participates in the regulation of many important physiological functions such as sleep-wakefulness cycle, motor coordination and neural plasticity, and cognition. However, as there are contradictory results regarding the melatonin production diurnal profile under alcohol consumption, the aim of this paper was to study the phenomenology and mechanisms of the putative modifications on the daily profile of melatonin production in rats submitted to chronic alcohol intake. The present results show that rats receiving 10% ethanol in drinking water for 35 days display an altered daily profile of melatonin production, with a phase delay and a reduction in the nocturnal peak. This can be partially explained by a loss of the daily rhythm and the 25% reduction in tryptophan hydroxylase activity and, mainly, by a phase delay in arylalkylamine N acetyltransferase gene expression and a 70% reduction in its peak activity. Upstream in the melatonin synthesis pathway, the results showed that noradrenergic signaling is impaired as well, with a decrease in beta1 and alpha1 adrenergic receptors' mRNA contents and in vitro sustained loss of noradrenergic stimulated melatonin production by glands from alcohol-treated rats. Together, these results confirm the alterations in the daily melatonin profile of alcoholic rats and suggest the possible mechanisms for the observed melatonin synthesis modification. PMID- 21635670 TI - Screening of patients with cognitive impairment when entering a rehabilitation unit. AB - AIM: The objective of this study is to evaluate the feasibility of an early screening of cognitive impairment in rehabilitation units, and the proportion of cognitive impairments (in particular, Alzheimer's disease) among this population. METHODS: This is a multicentric, non-interventional, epidemiological, transverse, projective and comprehensive study of five French rehabilitation units. RESULTS: This study on 279 people over 75 years found 105 new cases of cognitive impairments, among which 62 cases of dementia. CONCLUSION: These findings show the interest of generalizing cognitive impairments screening when entering a rehabilitation unit, in coordination with memory consultations and general practitioners. PMID- 21635671 TI - Antagonistic competition moderates virulence in Bacillus thuringiensis. AB - Classical models of the evolution of virulence predict that multiple infections should select for elevated virulence, if increased competitiveness arises from faster growth. However, diverse modes of parasite competition (resource-based, antagonism, immunity manipulation) can lead to adaptations with different implications for virulence. Using an experimental evolution approach we investigated the hypothesis that selection in mixed-strain infections will lead to increased antagonism that trades off against investment in virulence. Selection in mixed infections led to improved suppression of competitors in the bacterial insect pathogen Bacillus thuringiensis. Increased antagonism was associated with decreased virulence in three out of four selected lines. Moreover, mixed infections were less virulent than single-strain infections, and between-strain competition tended to decrease pathogen growth in vivo and in vitro. Spiteful interactions among these bacteria may be favoured because of the high metabolic costs of virulence factors and the high risk of mixed infections. PMID- 21635673 TI - Composition and dynamics of the gill microbiota of an invasive Indo-Pacific oyster in the eastern Mediterranean Sea. AB - Gill bacterial communities of Chama pacifica, an Indo-Pacific invasive oyster to the eastern Mediterranean Sea, were compared with those of Chama savignyi, its northern Red Sea congeneric species. Summer and winter bacterial populations were characterized and compared using 16S rDNA clone libraries, and seasonal population dynamics were monitored by automated ribosomal intergenic spacer analysis (ARISA). Clone libraries revealed a specific clade of bacteria, closely related to marine endosymbionts from the Indo-Pacific, found in both ecosystems, of which one taxon was conserved in oysters from both sites. This taxon was dominant in summer libraries and was weakly present in winter ones, where other members of this group were dominant. ARISA results revealed significant seasonal variation in bacterial populations of Mediterranean Sea oysters, as opposed to Red Sea ones that were stable throughout the year. We suggest that this conserved association between bacteria and oyster reflects either a symbiosis between the oyster host and some of its bacteria, a co-invasion of both parties, or both. PMID- 21635672 TI - Exploring and quantifying fungal diversity in freshwater lake ecosystems using rDNA cloning/sequencing and SSU tag pyrosequencing. AB - Water samples were collected along transects from the shore to the centre of two French lakes: the deep, volcanic, oligomesotrophic and low allochthonic-impacted Lake Pavin, and the productive and higher allochthonic-impacted Lake Aydat. The biodiversity was analysed using two approaches: the classical approach consisting of cloning/sequencing of the 18S, ITS1, 5.8S, ITS2 and partial 28S region using primers designed for fungus sequences, and the pyrosequencing of 18S rRNA hypervariable V2, V3 and V5 regions using two primer sets (one universal for eukaryotes and one for fungi). The classical approach yielded 146 (Lake Pavin) and 143 (Lake Aydat) sequences, corresponding to 46 and 63 operational taxonomic units (OTUs) respectively. Fungi represented half of the OTUs identified in Lake Pavin and 30% in Lake Aydat, and were dominated by sequences from Chytridiomycota found throughout Lake Pavin but mostly in the central pelagic zone of Lake Aydat. The pyrosequencing approach yielded 42,064 (Pavin) and 61,371 (Aydat) reads, of which 12-15% and 9-19% reads were assigned to fungi in Lakes Pavin and Aydat respectively. Chytridiomycota members were also dominant among these reads, with OTUs displaying up to > 33-fold overrepresentation in the centre compared with the riparian areas of Lake Aydat. Besides fungi, both approaches revealed other major eukaryote groups, with the highest diversity in the central areas of lakes. One of the major findings of our study was that the two lakes displayed contrasting spatial distributions, homogenous for Lake Pavin and heterogeneous for Lake Aydat, which may be related to their peculiarities. This study represents the first unveiling of microbial eukaryote and fungus diversity assessed with two complementary molecular methods, and is considered a major milestone towards understanding the dynamics and ecology of fungi in freshwater lake ecosystems, which are directly link to the abundance and distribution of taxa. PMID- 21635675 TI - Short-term metabolic effects of prednisone administration in healthy subjects. AB - AIMS: Supraphysiologic glucocorticoid activity is well established to cause impaired glucose tolerance and insulin resistance, yet no study has evaluated dose-dependent effects of low-dose prednisone during short-term oral administration. METHODS: The objective of this study was to quantify the effects of daily 10 or 25 mg prednisone administration for one week on insulin sensitivity by employing a two-step hyperinsulinemic euglycemic glucose clamp (Step 1: insulin infusion = 20 mU/m2/min; Step 2: insulin infusion = 80 mU/m2/min) in healthy, lean males. The amount of glucose infused at steady-state to maintain stable blood glucose [90 mg/dl (4.95 mmol/l)] was used to calculate several indices of insulin sensitivity. RESULTS: During Step 1 of the clamp, whole body glucose disposal (M) was reduced by 35% (p = 0.003) and M/I was reduced by 29% (p = 0.025) for 25 mg prednisone compared to placebo. No appreciable effect of 10 mg prednisone was observed. During Step 2, M was reduced by 33% (p = 0.001) and 15% (p = 0.006) for 25 and 10 mg prednisone compared to placebo; and M/I ratio was reduced by 31% (p < 0.001) and 13% (p = 0.026), respectively. The insulin sensitivity index, Si, calculated as the quotient of augmentation of M/I between Step 1 and 2, was reduced by 35.3% (p < 0.01) and 23.5% (p < 0.05) for 25 and 10 mg prednisone, respectively. CONCLUSION: Administration of relatively low pharmacological doses of prednisone for one week impaired insulin sensitivity in a dose-dependent manner in healthy males. These observed changes in insulin sensitivity are likely to be clinically relevant, especially in individuals predisposed to develop glucose intolerance. PMID- 21635674 TI - GLP-1 signals via ERK in peripheral nerve and prevents nerve dysfunction in diabetic mice. AB - AIM: Glucagon-like peptide-1 (GLP-1) is an incretin hormone that induces glucose dependent insulin secretion and may have neurotrophic properties. Our aim was to identify the presence and activity of GLP-1 receptors (GLP-1Rs) in peripheral nerve and to assess the impact of GLP-1R agonists on diabetes-induced nerve disorders. METHODS: Tissues were collected from streptozotocin-diabetic rats. GLP 1R function was assessed by incubating tissues from normal and diabetic rats with GLP-1R agonists and antagonists and measuring induction of ERK1/2 phosphorylation by Western blot. Streptozotocin-diabetic mice were also treated with the GLP-1R agonist exenatide for 8 weeks to assess the impact of GLP-1R signalling on peripheral nerve function and structure. RESULTS: GLP-1R protein was detected in rat dorsal root ganglia and the neurons and Schwann cells of the sciatic nerve. Protein levels were not affected by streptozotocin-induced diabetes. GLP-1R agonists did not signal via ERK1/2 in sciatic nerve of normal rats. However, GLP 1R agonists significantly increased pERK1/2 levels in sciatic nerves from diabetic rats, indicating that GLP-1Rs are functional in this tissue. Exenatide treatment did not affect blood sugar, insulin levels or paw thermal response latencies in either control or diabetic mice. However, the reductions of motor nerve conduction velocity and paw intraepidermal fibre density seen in diabetic mice were attenuated by exenatide treatment. CONCLUSIONS: These data show that the peripheral nerve of diabetic rodents exhibits functional GLP-1R and suggest that GLP-1R-mediated ERK-signalling in sciatic nerve of diabetic rodents may protect large motor fibre function and small C fibre structure by a mechanism independent of glycaemic control. PMID- 21635676 TI - Is impostor syndrome getting in the way of writing for the Journal of Forensic Nursing? PMID- 21635677 TI - Struggling to survive: the experiences of women sexually assaulted while intoxicated. AB - Approximately half of all sexual assault cases involve substance abuse or misuse, yet no studies have focused specifically on women who were under the influence of a substance when assaulted. The aim of this study was to explore the experiences of women who were sexually assault while under the influence of a substance. A phenomenological approach was used to gather data using individual, in-depth interviews with women following a sexual assault while intoxicated. Interviews were conducted using open-ended and probing questions to explore participants' life experiences from childhood to the present. The study uncovered a continuous struggle to survive among the participants. Five themes including previous victimization, substance misuse, struggling with feelings, finding support, and struggling to break the cycle were found. This research revealed lives complicated by substance misuse and histories of victimization. Intoxication raises vulnerability and risk for sexual assault. Effective nursing interventions designed to address the women's history of victimization and substance misuse may benefit women sexually assaulted while intoxicated. PMID- 21635678 TI - Working under threat: fear and nurse-patient interactions in a forensic psychiatric setting. AB - The purpose of this article is to present the results of a study conducted in a Canadian medium-security forensic psychiatric facility. The primary objective of this qualitative research was to describe and comprehend how fear influences nurse-patient interactions in a forensic psychiatric setting. Eighteen semistructured interviews with nurses were used as the primary source of data for analysis. In brief, the results from this research indicate, as other researchers have demonstrated, that within this highly regimented context, nurses are socialized to incorporate representations of the patients as being potentially dangerous, and, as a result, distance themselves from idealistic conceptions of care. Moreover, the research results emphasize the implication of fear in nurse patient interactions and particularly how fear reinforces nurses' need to create a safe environment in order to practice. A constant negotiation between space, "at risk" bodies and security takes place where nurses are forced to scrutinize their actions in order to avoid becoming victims of violence. In parallel, participants also described how being able to self-identify with patients enabled therapeutic interventions to take place. However, exposure to the patient's criminal history fostered negative reactions on the nurses' part, which impede nursing work. PMID- 21635679 TI - Reply to article "Receiving a forensic medical exam without participating in the criminal justice process: what will it mean?". AB - The Implications of the Violence Against Women Act (VAWA) pertaining to medical forensic exams are thoroughly explored in this analysis. The authors were motivated to write this article as a follow-up to a paper by Price (2010) previously published in the Journal of Forensic Nursing. Given the critical importance of this topic to forensic nursing, further examination of this issue is warranted, as a means of clarifying key issues, as well as addressing implications for the criminal justice and community response systems alike. PMID- 21635680 TI - Nonoccupational postexposure prophylaxis for human immunodeficiency virus in Sub Saharan Africa: a systematic review. AB - Both HIV and sexual assault remain major public health problems in Sub-Saharan Africa. This review examines the state of the science regarding provision of non- occupational post-exposure prophylaxis following sexual assault in Sub-Saharan African countries over the last 10 years. Specifically, rates of provision of HIV nPEP, the number of patients accepting nPEP, patients' rates of completing the HIV nPEP regimen as defined by the individual studies, and patients' perceptions of HIV nPEP from qualitative data. PMID- 21635681 TI - Criminalistics and the forensic nursing process. AB - Students learn science by actually performing science activities. The 12 laboratories described in this article assist students in applying the fundamental techniques germane to the field of forensic science to "solve" contrived cases and present "evidence" in a mock trial. Moreover, students are also confronted with some of the legal and ethical issues concerning the validity, reliability, and application of some forensic techniques. The pedagogical design of the laboratory course provides a rich, challenging, and interdisciplinary academic experience intended to augment and compliment the didactic forensic lecture portion of the course. This laboratory course was designed to engender, embody, and articulate one of the University's directive goals to support interdisciplinary teaching, research, and programming. Because we developed the laboratories on minimal funds, we demonstrated that it could be cost-effective. And thus, we recommend a laboratory science course be included as part of the curriculum of all forensic nursing students and practitioners. PMID- 21635682 TI - Consumerism and its impact on robotic-assisted radical prostatectomy. AB - OBJECTIVES: * Many experts consider that media coverage, marketing and/or direct to-consumer advertising, particularly Internet-based forms, are fundamental to the widespread adoption of robotic-assisted prostatectomy (RARP). However, this has not been explored previously. * The primary objective of the present study was to delineate the role of media coverage and marketing of RARP on the Internet, whereas the secondary goal focused on website quality with respect to the presentation of prostatectomy. MATERIALS AND METHODS: * Website content was evaluated for direct-to-consumer advertising after the retrieval of the first 50 websites using Google and Yahoo for each of the terms: 'robotic prostatectomy, laparoscopic prostatectomy (LP) and open radical prostatectomy (ORP)'. * A linear regression analysis was performed for the annual number of Internet news hits over the last decade for each procedure. Website quality assessment was performed using WHO Honesty on the Internet (HON) code principles. RESULTS: * Of the retrieved sites, the proportion containing direct-to-consumer advertising for RARP vs LP vs ORP using Google was 64% vs 14% vs 0%, respectively (P < 0.001) and, using Yahoo, 80% vs 16% vs 0%, respectively (P < 0.001). * In a linear regression analysis, the r(2) values for news hits for each year over the last 10 years were 0.89, 0.74 and 0.76 for RARP, LP and ORP, respectively. * Website quality assessment found that a minority of the websites were accredited with HONcode principles, with no difference between procedure types (P > 0.05). CONCLUSIONS: * Media coverage and marketing of RARP on the Internet is more widespread compared to LP and ORP. * Disturbingly, the quality of websites using any technique for prostatectomy was of poor quality when using principles of honest information presenting and such findings need to be discussed with respect to obtaining informed consent from patients. PMID- 21635683 TI - Twenty-year prevalence of diabetes mellitus and hypertension in patients receiving shock-wave lithotripsy for urolithiasis. AB - OBJECTIVES: To compare the prevalence of hypertension and diabetes mellitus (DM) in patients treated with an unmodified HM-3 lithotripter (USWL) and a second generation modified HM-3 lithotripter (MSWL) 20 years ago at our Centre with that in the provincial population. To determine whether the type of lithotripter was differentially associated with the development of these sequelae. PATIENTS AND METHODS: Retrospective review of 727 patients at Vancouver General Hospital who underwent shock-wave lithotripsy (SWL) between 1985 and 1989. Our study group was compared with Statistics Canada data describing the provincial prevalence of these diseases. Multivariate analysis was performed. RESULTS: The response rate was 37.3%. There was a greater proportion of overweight and obese individuals in the study group compared with the provincial average. In univariate analysis, lithotripsy with an unmodified HM-3 (USWL) was associated with a higher rate of DM than the provincial rate, whereas lithotripsy with the modified HM-3 (MSWL) was not. Hypertension was more prevalent in all lithotripsy subjects. On multivariate analysis the type of lithotripter was not associated with the development of either sequela. CONCLUSIONS: No association between lithotripsy and the development of either DM or hypertension in a multivariate analysis Metabolic syndrome may have elevated the prevalence of DM and hypertension observed in our subjects on univariate analysis, which is in keeping with the fact that our study population had statistically higher body mass indices than the provincial rate. Lithotripsy using the HM-3 was not associated with increased DM or hypertension. PMID- 21635684 TI - Carbon storage in terrestrial ecosystems: do browsing and grazing herbivores matter? AB - Large mammalian herbivores manifest a strong top-down control on ecosystems that can transform entire landscapes, but their impacts have not been reviewed in the context of terrestrial carbon storage. Here, we evaluate the effects of plant biomass consumption by large mammalian herbivores (>10 kg adult biomass), and the responses of ecosystems to these herbivores, on carbon stocks in temperate and tropical regions, and the Arctic. We calculate the difference in carbon stocks resulting from herbivore exclusion using the results of 108 studies from 52 vegetation types. Our estimates suggest that herbivores can reduce terrestrial above- and below-ground carbon stocks across vegetation types but reductions in carbon stocks may approach zero given sufficient periods of time for systems to respond to herbivory (i.e. decades). We estimate that if all large herbivores were removed from the vegetation types sampled in our review, increases in terrestrial carbon stocks would be up to three orders of magnitude less than many of the natural and human-influenced sources of carbon emissions. However, we lack estimates for the effects of herbivores on below-ground biomass and soil carbon levels in many regions, including those with high herbivore densities, and upwards revisions of our estimates may be necessary. Our results provide a starting point for a discussion on the magnitude of the effects of herbivory on the global carbon cycle, particularly given that large herbivores are common in many ecosystems. We suggest that herbivore removal might represent an important strategy towards increasing terrestrial carbon stocks at local and regional scales within specific vegetation types, since humans influence populations of most large mammals. PMID- 21635685 TI - Total intravenous anaesthesia by boluses or by continuous rate infusion of propofol in mute swans (Cygnus olor). AB - OBJECTIVE: To investigate intravenous (IV) propofol given by intermittent boluses or by continuous rate infusion (CRI) for anaesthesia in swans. STUDY DESIGN: Prospective randomized clinical study. ANIMALS: Twenty mute swans (Cygnus olor) (eight immature and 12 adults) of unknown sex undergoing painless diagnostic or therapeutic procedures. METHODS: Induction of anaesthesia was with 8 mg kg(-1) propofol IV. To maintain anaesthesia, ten birds (group BOLI) received propofol as boluses, whilst 10 (group CRI) received propofol as a CRI. Some physiological parameters were measured. Anaesthetic duration was 35 minutes. Groups were compared using Mann-Whitney U-test. Results are median (range). RESULTS: Anaesthetic induction was smooth and tracheal intubation was achieved easily in all birds. Bolus dose in group BOLI was 2.9 (1.3-4.3) mg kg(-1); interval between and number of boluses required were 4 (1-8) minutes and 6 (4-11) boluses respectively. Total dose of propofol was 19 (12.3-37.1) mg kg(-1). Awakening between boluses was very abrupt. In group CRI, propofol infusion rate was 0.85 (0.8-0.9) mg kg(-1) minute(-1), and anaesthesia was stable. Body temperature, heart and respiratory rates, oxygen saturation (by pulse oximeter) and reflexes did not differ between groups. Oxygen saturations (from pulse oximeter readings) were low in some birds. Following anaesthesia, all birds recovered within 40 minutes. In 55% of all, transient signs of central nervous system excitement occurred during recovery. CONCLUSIONS AND CLINICAL RELEVANCE: 8 mg kg(-1) propofol appears an adequate induction dose for mute swans. For maintenance, a CRI of 0.85 mg kg(-1) minute(-1) produced stable anaesthesia suitable for painless clinical procedures. In contrast bolus administration, was unsatisfactory as birds awoke very suddenly, and the short intervals between bolus requirements hampered clinical procedures. Administration of additional oxygen throughout anaesthesia might reduce the incidence of low arterial haemoglobin saturation. PMID- 21635686 TI - IL-15 preferentially enhances functional properties and antigen-specific responses of CD4+CD28(null) compared to CD4+CD28+ T cells. AB - One of the most prominent changes during T-cell aging in humans is the accumulation of CD28(null) T cells, mainly CD8+ and also CD4+ T cells. Enhancing the functional properties of these cells may be important as they provide an antigen-specific defense against chronic infections. Recent studies have shown that IL-15 does in fact play an appreciable role in CD4 memory T cells under physiological conditions. We found that treatment with IL-15 increased the frequency of elderly CD4+CD28(null) T cells by the preferential proliferation of these cells compared to CD4+CD28+ T cells. IL-15 induced an activated phenotype in CD4+CD28(null) T cells. Although the surface expression of IL-15R alpha-chain was not increased, the transcription factor STAT-5 was preferentially activated. IL-15 augmented the cytotoxic properties of CD4+CD28(null) T cells by increasing both the mRNA transcription and storage of granzyme B and perforin for the cytolytic effector functions. Moreover, pretreatment of CD4+CD28(null) T cells with IL-15 displayed a synergistic effect on the IFN-gamma production in CMV specific responses, which was not observed in CD4+CD28+ T cells. IL-15 could play a role enhancing the effector response of CD4+CD28(null) T cells against their specific chronic antigens. PMID- 21635687 TI - Scalene muscle injections for neurogenic thoracic outlet syndrome: case series. AB - Scalene muscle injections are used to confirm the diagnosis of neurogenic thoracic outlet syndrome and predict the response of patients to surgery. We performed a retrospective study to determine if relief of pain was related to brachial plexus blockade in these patients. METHODS: We reviewed the charts of 12 patients who had anterior and middle scalene muscle injections, for neurogenic thoracic outlet syndrome, between April 2009 and September 2010. The injections were performed under ultrasound guidance wherein 2 to 5 mL of 0.25% bupivacaine was injected into the belly of the anterior and scalene muscles. The following were noted: (1) sites of preprocedure pain; (2) volume injected into each of the anterior and middle scalene muscles; (3) presence of numbness after injection; and (4) presence and duration of pain relief. RESULTS: All 12 patients had relief of their pain. Six of the twelve patients developed numbness, which ranged from blockade of the C4-5, C6-7, and C4-T1 dermatomes. In the patients who developed numbness, there was no relationship between the duration of numbness and the duration of pain relief or the location of numbness and the location of pain relief. CONCLUSIONS: The relief from scalene muscle injections in patients with neurogenic thoracic outlet syndrome is not related to blockade of the brachial plexus. PMID- 21635688 TI - Overexpression of human calnexin in yeast improves measles surface glycoprotein solubility. AB - The limitations of high-level expression of virus surface proteins in yeast are not well understood. The inefficiency of yeast to produce active human virus surface glycoproteins, as well as other mammalian glycoproteins, is usually explained by the inefficient folding of the glycoprotein into its characteristic and functional three-dimensional structure from a random coil. The endoplasmic reticulum (ER) is a highly versatile protein factory that is equipped with chaperones and folding enzymes essential for protein folding. To improve folding and solubility of viral surface glycoprotein, the genes encoding human ER resident chaperones calnexin, calreticulin, immunoglobin binding protein (BiP), protein disulfide isomerase (PDI) and foldase (ERp57) were coexpressed together with hemagglutinin gene from measles virus in the yeast Saccharomyces cerevisiae. The effect of coexpressing chaperones on the total yield of measles virus hemagglutinin (MeH) as well as the intracellular fate of the glycoprotein was determined. Our results demonstrated that coexpression of human calnexin noticeably enhanced the quantity of the soluble glycosylated form of MeH in yeast. The coexpression of human calreticulin-, PDI-, ERp57- and BiP-encoding genes did not improve the quality of recombinant MeH. PMID- 21635689 TI - Avulsion of permanent teeth: theory to practice. AB - This paper presents clinical protocols for the emergency, early and post treatment complications of the avulsed tooth. The biological basis for these protocols is presented so that the reader understands the clinical decisions that have been made. Most of the protocols described in this article, but not all, have been adopted in the official guidelines of the International Association of Dental Traumatology. Some experimental results are promising and they have therefore been included in the review to stimulate colleagues to further research. PMID- 21635690 TI - Surfing for mouth guards: assessing quality of online information. AB - INTRODUCTION: The Internet is an easily accessible and commonly used source of health-related information, but evaluations of the quality of this information within the dental trauma field are still lacking. AIM: The aims of this study are (i) to present the most current scientific knowledge regarding mouth guards used in sport activities, (ii) to suggest a scoring system to evaluate the quality of information pertaining to mouth guard protection related to World Wide Web sites and (iii) to employ this scoring system when seeking reliable mouth guard-related websites. MATERIALS AND METHODS: First, an Internet search using the keywords 'athletic injuries/prevention and control' and 'mouth protector' or 'mouth guards' in English was performed on PubMed, Cochrane, SvedMed+ and Web of Science to identify scientific knowledge about mouth guards. Second, an Internet search using the keywords 'consumer health information Internet', 'Internet information public health' and 'web usage-seeking behaviour' was performed on PubMed and Web of Science to obtain scientific articles seeking to evaluate the quality of health information on the Web. Based on the articles found in the second search, two scoring systems were selected. Then, an Internet search using the keywords 'mouth protector', 'mouth guards' and 'gum shields' in English was performed on the search engines Google, MSN and Yahoo. The websites selected were evaluated for reliability and accuracy. RESULTS: Of the 223 websites retrieved, 39 were designated valid and evaluated. Nine sites scored 22 or higher. The mean total score of the 39 websites was 14.2. Fourteen websites scored higher than the mean total score, and 25 websites scored less. The highest total score, presented by a Public Institution Web site (Health Canada), was 31 from a maximum possible score of 34, and the lowest score was 0. CONCLUSION: This study shows that there is a high amount of information about mouth guards on the Internet but that the quality of this information varies. It should be the responsibility of health care professionals to suggest and provide reliable Internet URL addresses to patients. In addition, an appropriate search terminology and search strategy should be made available to persons who want to search beyond the recommended sites. PMID- 21635691 TI - Pattern of maxillofacial fractures at a tertiary hospital in northern India: a 4 year retrospective study of 718 patients. AB - PURPOSE: Pattern and aetiology of maxillofacial injuries varies from one country to another and even within the same country depending on prevailing socio economic, cultural and environmental factors. Various studies have been carried out in various countries to study the epidemiology and demographics of the maxillofacial injuries but the studies from India are few. MATERIAL AND METHODS: Retrospective study was undertaken at Christian Medical College Ludhiana (India) from January 2006 to December 2009. Treatment records of the patients was checked and age, gender, aetiology of injury, associated injuries, maxillofacial fractures and treatment offered were recorded. RESULTS: A total of 1075 fractures were recorded in 718 patients, ranging from 11 months to 85 years of age. Male:female ratio was 6.6:1. Maxillofacial injuries were most common in third decade of life. A total of 517 patients suffered injuries because of road traffic accident, 115 because of accidental fall and 67 because of interpersonal violence. A total of 184 patients had 221 associated injuries of which 56.1% head injuries, 29.0% orthopaedic injuries and 14.9% other injuries were present. Of 596 middle third fractures, 29.8% were managed conservatively; for 21.7% and 48.7% of fractures, closed reduction and open reduction were performed respectively. Of 479 mandibular fractures, 1.8% was managed conservatively; for 16.7% and 81.5% fractures, closed reduction and open reduction were performed respectively. CONCLUSION: Better socio-economic status of people, increased vehicular movements and non-implementation of road safety norms have increased road traffic accidents. Proper education of the people who are most commonly involved in the RTA can be one of the possible ways to reduce the maxillofacial injuries. PMID- 21635693 TI - Hyperactive behaviour in the mouse model of mucopolysaccharidosis IIIB in the open field and home cage environments. AB - Mucopolysaccharidosis IIIB (MPS IIIB) is a lysosomal storage disorder characterized by severe behavioural disturbances and progressive loss of cognitive and motor function. There is no effective treatment, but behavioural testing is a valuable tool to assess neurodegeneration and the effect of novel therapies in mouse models of disease. Several groups have evaluated behaviour in this model, but the data are inconsistent, often conflicting with patient natural history. We hypothesize that this discrepancy could be due to differences in open field habituation and home cage behaviour. Eight-month-old wild-type and MPS IIIB mice were tested in a 1-h open field test, performed 1.5 h after lights on, and a 24-h home cage behaviour test performed after 24 h of acclimatization. In the 1-h test, MPS IIIB mice were hyperactive, with increased rapid exploratory behaviour and reduced immobility time. No differences in anxiety were seen. Over the course of the test, differences became more pronounced with maximal effects at 1 h. The 24-hour home cage test was less reliable. There was evidence of increased hyperactivity in MPS IIIB mice, however, immobility was also increased, suggesting a level of inconsistency in this test. Performance of open field analysis within 1-2 h after lights on is probably critical to achieving maximal success as MPS IIIB mice have a peak in activity around this time. The open field test effectively identifies hyperactive behaviour in MPS IIIB mice and is a significant tool for evaluating effects of therapy on neurodegeneration. PMID- 21635694 TI - Flavogenomics--a genomic and structural view of flavin-dependent proteins. AB - Riboflavin (vitamin B(2)) serves as the precursor for FMN and FAD in almost all organisms that utilize the redox-active isoalloxazine ring system as a coenzyme in enzymatic reactions. The role of flavin, however, is not limited to redox processes, as ~ 10% of flavin-dependent enzymes catalyze nonredox reactions. Moreover, the flavin cofactor is also widely used as a signaling and sensing molecule in biological processes such as phototropism and nitrogen fixation. Here, we present a study of 374 flavin-dependent proteins analyzed with regard to their function, structure and distribution among 22 archaeal, eubacterial, protozoan and eukaryotic genomes. More than 90% of flavin-dependent enzymes are oxidoreductases, and the remaining enzymes are classified as transferases (4.3%), lyases (2.9%), isomerases (1.4%) and ligases (0.4%). The majority of enzymes utilize FAD (75%) rather than FMN (25%), and bind the cofactor noncovalently (90%). High-resolution structures are available for about half of the flavoproteins. FAD-containing proteins predominantly bind the cofactor in a Rossmann fold (~ 50%), whereas FMN-containing proteins preferably adopt a (betaalpha)(8)-(TIM)-barrel-like or flavodoxin-like fold. The number of genes encoding flavin-dependent proteins varies greatly in the genomes analyzed, and covers a range from ~ 0.1% to 3.5% of the predicted genes. It appears that some species depend heavily on flavin-dependent oxidoreductases for degradation or biosynthesis, whereas others have minimized their flavoprotein arsenal. An understanding of 'flavin-intensive' lifestyles, such as in the human pathogen Mycobacterium tuberculosis, may result in valuable new intervention strategies that target either riboflavin biosynthesis or uptake. PMID- 21635695 TI - Stereoselectivity and conformational stability of haloalkane dehalogenase DbjA from Bradyrhizobium japonicum USDA110: the effect of pH and temperature. AB - The effect of pH and temperature on structure, stability, activity and enantioselectivity of haloalkane dehalogenase DbjA from Bradyrhizobium japonicum USDA110 was investigated in this study. Conformational changes have been assessed by circular dichroism spectroscopy, functional changes by kinetic analysis, while quaternary structure was studied by gel filtration chromatography. Our study shows that the DbjA enzyme is highly tolerant to pH changes. Its secondary and tertiary structure was not affected by pH in the ranges 5.3-10.3 and 6.2-10.1, respectively. Oligomerization of DbjA was strongly pH-dependent: monomer, dimer, tetramer and a high molecular weight cluster of the enzyme were distinguished in solution at different pH conditions. Moreover, different oligomeric states of DbjA possessed different thermal stabilities. The highest melting temperature (T(m) = 49.1 +/- 0.2 degrees C) was observed at pH 6.5, at which the enzyme occurs in dimeric form. Maximal activity was detected at 50 degrees C and in the pH interval 7.7-10.4. While pH did not have any effect on enantiodiscriminination of DbjA, temperature significantly altered DbjA enantioselectivity. A decrease in temperature results in significantly enhanced enantioselectivity. The temperature dependence of DbjA enantioselectivity was analysed with 2-bromobutane, 2 bromopentane, methyl 2-bromopropionate and ethyl 2-bromobutyrate, and differential activation parameters Delta(R-S)DeltaH and Delta(R-S)DeltaS were determined. The thermodynamic analysis revealed that the resolution of beta bromoalkanes was driven by both enthalpic and entropic terms, while the resolution of alpha-bromoesters was driven mainly by an enthalpic term. Unique catalytic activity and structural stability of DbjA in a broad pH range, combined with high enantioselectivity with particular substrates, make this enzyme a very versatile biocatalyst. Enzyme EC3.8.1.5 haloalkane dehalogenase. PMID- 21635696 TI - Observing stage-shifts in head and neck squamous cell carcinoma from initial clinical outpatient staging to definite clinical tumour board staging using radiological and endoscopical investigations: will less do? AB - OBJECTIVES: Staging head and neck squamous cell carcinoma usually is initiated in the outpatient department (OPD) using patient history and physical examination. To reach consensus on stage and therapy, imaging and panendoscopy are more or less routinely applied as additional staging tests. Staging results from the outpatient department were evaluated for tumour and neck stage. We investigated in which situations additional staging tests are needed. DESIGN: Prospective cohort study. SETTING: Head and Neck Oncology Group, University Medical Centre. PARTICIPANTS: Of 341 patients, initial staging results from the outpatient department were compared with the ultimate staging results from the tumour board review, which provides the basis for decisions on therapy. MAIN OUTCOME MEASURES: Tumour stage and neck stage migrations during this staging tract. RESULTS: In staging head and neck tumours are squamous cell carcinomas, additional staging tests like imaging and/or panendoscopy remain necessary to evaluate the primary tumour stage. Average accuracies of T-staging in oral cavity, pharynx and larynx only amount to 71%, 47% and 61%, respectively. For T1N0 and T2N0 oral cavity tumours, neck assessment in the outpatient department was accurate in 100% (95% CI: 91-100%) and 93% (95% CI: 80-98%), respectively. In the larynx group, this was 100% (95% CI: 91-100%) and 97% (95% CI: 86-100%), respectively. CONCLUSIONS: For oral cavity and laryngeal tumours staged as T1-2N0 in the outpatient department, we concluded that the outpatient department staging is highly predictive of the final pre-treatment staging. In these cases, computer tomography and/or MRI remain a necessity but additional ultrasound with or without fine needle aspiration cytology can be omitted. In our institution, this would have resulted in a 46% reduction in ultrasound procedures. If T-stage is upgraded during imaging or panendoscopy, additional staging tests for N-stage should still be considered. PMID- 21635697 TI - A simple and accurate method to sex savannah, forest and Asian elephants using noninvasive sampling techniques. AB - We report the development of a reliable and efficient method for molecular sexing of all extant elephant taxa. We developed primers that amplify two short Y specific fragments (SRY1 and AMELY2) and one longer X-specific fragment (PLP1), developed from elephant sequences in one multiplex PCR. All fragments were designed to be short (< 200 basepairs) for use with degraded DNA and to be 50 basepairs apart to optimize visualization on agarose gels or as electropherograms. The multiplex PCR method matched sexes for at least 97.9% of the noninvasive savannah elephant samples and produced the expected female/male banding patterns for 14 African forest and 11 Asian elephant samples. We found this method to be more robust, efficient and less prone to contamination than previously developed sexing methods for elephants. PMID- 21635698 TI - Comparing and combining distance-based and character-based approaches for barcoding turtles. AB - Molecular barcoding can serve as a powerful tool in wildlife forensics and may prove to be a vital aid in conserving organisms that are threatened by illegal wildlife trade, such as turtles (Order Testudines). We produced cytochrome oxidase subunit one (COI) sequences (650 bp) for 174 turtle species and combined these with publicly available sequences for 50 species to produce a data set representative of the breadth of the order. Variability within the barcode region was assessed, and the utility of both distance-based and character-based methods for species identification was evaluated. For species in which genetic material from more than one individual was available (n = 69), intraspecific divergences were 1.3% on average, although divergences greater than the customary 2% barcode threshold occurred within 15 species. High intraspecific divergences could indicate species with a high degree of internal genetic structure or possibly even cryptic species, although introgression is also probable in some of these taxa. Divergences between species of the same genus were 6.4% on average; however, 49 species were <2% divergent from congeners. Low levels of interspecific divergence could be caused by recent evolutionary radiations coupled with the low rates of mtDNA evolution previously observed in turtles. Complementing distance-based barcoding with character-based methods for identifying diagnostic sets of nucleotides provided better resolution in several cases where distance-based methods failed to distinguish species. An online identification engine was created to provide character-based identifications. This study constitutes the first comprehensive barcoding effort for this seriously threatened order. PMID- 21635699 TI - Diagnosis of tuberculosis in camelids: old problems, current solutions and future challenges. AB - In spite of great efforts for its control and eradication, tuberculosis remains one of the most important zoonosis worldwide. Its causative agents, the members of the Mycobacterium tuberculosis complex, have a wide host range that complicates the epidemiology of this disease. Among susceptible species to these pathogens, camelids from the New World (llama, alpaca and vicuna) and Old World (Bactrian camel and dromedary) are acquiring an increasing importance in several European countries because of its growing number and could act as reservoirs of the disease for livestock and humans in their natural habitat. In addition, tuberculosis caused by a number of M. tuberculosis complex members is a life threatening disease in these animal species. Although tuberculosis has been known to affect camelids for a long time, ante-mortem diagnosis is still challenging because of the lack of standardized diagnostic techniques and the limited sensitivity and specificity of the most widely applied tests. However, in recent years, several techniques that can at least partially overcome these limitations have been developed. This paper reviews the results and advances achieved in tuberculosis diagnosis in camelids in the last decade as well as the progresses on ongoing investigations, with special attention to the remaining challenges that still have to be faced to assure the availability of reliable tools for the detection of tuberculosis-infected animals and herds. PMID- 21635700 TI - Greening critical care. AB - Climate change and environmental stewardship are phrases that have been defining the past few decades and promoting change in our societies. The sensitivities of intensive care as a specialty make the process of greening an intensive care unit a challenge, but not one that is insurmountable. This paper discusses opportunities for critical care to reduce its environmental impact and provide a framework change. The article includes suggestions of what can be done as an individual, as a unit and as a hospital. Generally, practices in critical care are accepted without questioning the environmental consequences. We believe it is time for change, and critical care should give environmental stewardship a higher priority. PMID- 21635701 TI - Functional markers to predict the need for prolonged mechanical ventilation in patients with Guillain-Barre syndrome. PMID- 21635702 TI - Diverse perspectives on the current state of genomic medicine: has the revolution begun? AB - A report on the Future of Genomic Medicine IV meeting held in La Jolla, California, USA, 3-4 March 2011. PMID- 21635703 TI - Using lung ultrasound to differentiate patients in acute dyspnea in the prehospital emergency setting. AB - The diagnosis and treatment of dyspnea in the emergency department and in the prehospital setting is a challenge faced by the emergency physician and other prehospital care providers. While the use of lung ultrasound as a diagnostic tool in dyspneic patients has been well researched, there has been limited evaluation of its use in the prehospital setting. In the previous issue of Critical Care, Prosen and colleagues study the accuracy of lung ultrasound compared with both N terminal pro-brain natriuretic peptide and the clinical examination for differentiating between acute decompensated congestive heart failure and chronic obstructive pulmonary disease exacerbations for patients in the prehospital setting. Their article adds to the growing body of evidence demonstrating the diagnostic efficacy of lung ultrasound in differentiating between these two disease processes in the acutely dyspneic patient. PMID- 21635704 TI - The conundrum of persistent inappropriate use of frozen plasma. AB - Frozen plasma (FP) is commonly used for the treatment of bleeding or the prevention of bleeding in critically ill patients, but clinical evidence to help aid the critical care clinician make decisions on whether to transfuse or not is at present limited. Despite the limited evidence, it appears FP is administered not infrequently in the absence of bleeding or with no required procedure when the international normalized ratio (INR) is essentially normal (<1.5) or only mildly deranged (<2.5). The study by Stanworth and colleagues in a recent issue of Critical Care raises awareness of FP transfusion use in the critically ill, should prompt a consideration of curbing its use when it is not clearly appropriate, and illustrates the need for future high quality evidence to guide FP use in the critically ill when the risk:benefit ratio is less clear. PMID- 21635705 TI - Emergency overcrowding: an incurable disease? PMID- 21635706 TI - From bone to breast and back - the bone cytokine RANKL and breast cancer. AB - Receptor activator of nuclear factor-kappaB ligand (RANKL) plays a pivotal role in regulating bone homeostasis. Osteoporosis and malignant bone disease secondary to breast cancer are characterized by enhanced RANKL production and increased bone turnover. Thus, denosumab, a monoclonal antibody to RANKL, has been developed and is now approved for various bone loss conditions. Recent results indicate that RANKL may also promote the development and osseous migration of breast cancer. PMID- 21635707 TI - Who is in the driver's seat in 8p12 amplifications? ZNF703 in luminal B breast tumors. AB - Two recent reports identify ZNF703 as an oncogene driving selection of frequent chromosome 8p12 amplifications in luminal B breast tumors. The estrogen responsive ZNF703 gene encodes a transcriptional cofactor that, when overexpressed, induces cell proliferation and interferes with transforming growth factor beta signaling. In MCF7 cells, increased ZNF703 expression results in activation of genes involved in stem cell self-renewal - while in primary human mammary epithelial cells, ZNF703 increases the ratio of luminal to basal progenitors. Expression of the murine homolog of ZNF703 reduces cell adhesion and promotes metastasis. ZNF703 overexpression thus alters regulation of proliferation and differentiation in luminal B tumors. PMID- 21635708 TI - Jekyll or Hyde: does Matrigel provide a more or less physiological environment in mammary repopulating assays? AB - In vivo transplantation is the current 'gold-standard' assay for evaluating mammary stem cell (MaSC) function. Matrigel, a reconstituted extracellular matrix derived from a mouse sarcoma line, is increasingly being utilized for mammary repopulating assays, although original studies were carried out in its absence. This matrix has also been shown to enhance tumor-initiating capacity. Whilst Matrigel increases the rate of engraftment by MaSCs, it also appears to promote progenitor activity that is distinct from bona fide stem cell activity. This caveat should be considered when interpreting mammary reconstitution assays that incorporate Matrigel, particularly when transplanting high cell numbers. PMID- 21635710 TI - Personalizing carbamazepine therapy. AB - The anticonvulsant carbamazepine has a high incidence of cutaneous adverse drug reactions. A recent prospective clinical trial in Taiwan has indicated that HLA B*1502 screening will reduce the incidence of life-threatening adverse reactions to carbamazepine, while a genome-wide association study has identified the HLA A*3101 allele as a genetic risk factor for the full spectrum of carbamazepine induced cutaneous adverse drug reactions in a European population. These studies should aid future decision-making for personalized use of carbamazepine treatment. PMID- 21635711 TI - Massively parallel rare disease genetics. AB - A report on the 'Genomic Disorders 2011 - The Genomics of Rare Diseases' meeting, Wellcome Trust Sanger Institute, Hinxton, UK, 23-26 March 2011. PMID- 21635712 TI - A mystery unveiled. AB - A recent phylogenomic study has provided new evidence for two ancient whole genome duplications in plants, with potential importance for the evolution of seed and flowering plants. PMID- 21635713 TI - Mapping of disease-associated variants in admixed populations. AB - A more refined estimation of ancestry would benefit admixture mapping and association mapping, making disease loci identification in admixed populations more powerful. PMID- 21635714 TI - The volatile microbiome. AB - The first detailed temporal study of the human microbiome shows that individual body habitats exhibit surprising variation over time yet maintain distinguishable community structures. PMID- 21635715 TI - In praise of model organisms. PMID- 21635709 TI - Interpreting Breast International Group (BIG) 1-98: a randomized, double-blind, phase III trial comparing letrozole and tamoxifen as adjuvant endocrine therapy for postmenopausal women with hormone receptor-positive, early breast cancer. AB - The Breast International Group (BIG) 1-98 study is a four-arm trial comparing 5 years of monotherapy with tamoxifen or with letrozole or with sequences of 2 years of one followed by 3 years of the other for postmenopausal women with endocrine-responsive early invasive breast cancer. From 1998 to 2003, BIG -98 enrolled 8,010 women. The enhanced design f the trial enabled two complementary analyses of efficacy and safety. Collection of tumor specimens further enabled treatment comparisons based on tumor biology. Reports of BIG 1-98 should be interpreted in relation to each individual patient as she weighs the costs and benefits of available treatments. Clinicaltrials.gov ID: NCT00004205. PMID- 21635716 TI - Aspects of innate immunity in Sjogren's syndrome. AB - Previously, a dominant role of the adaptive immune system in the pathogenesis of Sjogren's syndrome was suspected. Recent advances, however, have revealed a major role of the type I IFN pathway, documented by an increased circulating type I IFN activity and an IFN 'signature' in peripheral blood mononuclear cells and minor salivary gland biopsies from the patients. Polymorphisms in the genes IRF5 and STAT4 leading to increased IFN activation are associated with disease susceptibility. In the pathogenesis of Sjogren's syndrome, the activation of salivary gland epithelial cells appears to be the initial event. Once intrinsically activated, they express costimulatory and Toll-like receptors (TLRs) and MHC class I and II molecules, can present autoantigens and produce proinflammatory cytokines. The subsequent activation of plasmacytoid dendritic cells induces the production of high levels of proinflammatory cytokines in individuals with the risk alleles of the susceptibility genes IRF5 and STAT4. Under the influence of the high IFN concentration in the glands and through TLR ligation, B-cell activating factor is produced by epithelial cells and, together with autoantigen presentation on salivary gland epithelial cells, stimulates the adaptive immune system. In view of the central role of IFNalpha in at least the initiation of the pathogenesis of Sjogren's syndrome, blockade of this cytokine may be a rational therapeutic approach. PMID- 21635718 TI - Regulation of bone by the adaptive immune system in arthritis. AB - Studies on the immune regulation of osteoclasts in rheumatoid arthritis have promoted the new research field of 'osteoimmunology', which investigates the interplay between the skeletal and immune systems at the molecular level. Accumulating evidence lends support to the theory that bone destruction associated with rheumatoid arthritis is caused by the enhanced activity of osteoclasts, resulting from the activation of a unique helper T cell subset, 'Th17 cells'. Understanding the interaction between osteoclasts and the adaptive immune system in rheumatoid arthritis and the molecular mechanisms of Th17 development will lead to the development of potentially effective therapeutic strategies. PMID- 21635717 TI - MicroRNA biogenesis and regulation of bone remodeling. AB - MicroRNAs (miRNAs) are key post-transcriptional regulators of gene expression. This review will highlight our current understanding of miRNA biogenesis and mechanisms of action, and will summarize recent work on the role of miRNAs, including the miR-29 family, in bone remodeling. These studies represent the first steps in demonstrating the importance of miRNAs in the control of osteoblast and osteoclast differentiation and function. An in-depth understanding of the roles of these regulatory RNAs in the skeleton will be critical for the development of new therapeutics aimed at treating bone loss and perhaps facilitating fracture repair. PMID- 21635719 TI - A network of SCOP hidden Markov models and its analysis. AB - BACKGROUND: The Structural Classification of Proteins (SCOP) database uses a large number of hidden Markov models (HMMs) to represent families and superfamilies composed of proteins that presumably share the same evolutionary origin. However, how the HMMs are related to one another has not been examined before. RESULTS: In this work, taking into account the processes used to build the HMMs, we propose a working hypothesis to examine the relationships between HMMs and the families and superfamilies that they represent. Specifically, we perform an all-against-all HMM comparison using the HHsearch program (similar to BLAST) and construct a network where the nodes are HMMs and the edges connect similar HMMs. We hypothesize that the HMMs in a connected component belong to the same family or superfamily more often than expected under a random network connection model. Results show a pattern consistent with this working hypothesis. Moreover, the HMM network possesses features distinctly different from the previously documented biological networks, exemplified by the exceptionally high clustering coefficient and the large number of connected components. CONCLUSIONS: The current finding may provide guidance in devising computational methods to reduce the degree of overlaps between the HMMs representing the same superfamilies, which may in turn enable more efficient large-scale sequence searches against the database of HMMs. PMID- 21635720 TI - Purification of functional baculovirus particles from silkworm larval hemolymph and their use as nanoparticles for the detection of human prorenin receptor (PRR) binding. AB - BACKGROUND: Baculovirus, which has a width of 40 nm and a length of 250-300 nm, can display functional peptides, receptors and antigens on its surface by their fusion with a baculovirus envelop protein, GP64. In addition, some transmembrane proteins can be displayed without GP64 fusion, using the native transmembrane domains of the baculovirus. We used this functionality to display human prorenin receptor fused with GFPuv (GFPuv-hPRR) on the surface of silkworm Bombyx mori nucleopolyhedrovirus (BmNPV) and then tested whether these baculovirus particles could be used to detect protein-protein interactions. RESULTS: BmNPV displaying GFPuv-hPRR (BmNPV-GFPuv-hPRR) was purified from hemolymph by using Sephacryl S 1000 column chromatography in the presence of 0.01% Triton X-100. Its recovery was 86% and the final baculovirus particles number was 4.98*108 pfu. Based on the results of enzyme-linked immunosorbent assay (ELISA), 3.1% of the total proteins in BmNPV-GFPuv-hPRR were GFPuv-hPRR. This value was similar to that calculated from the result of western blot by a densitometry (2.7%). To determine whether BmNPV-GFPuv-hPRR particles were bound to human prorenin, ELISA results were compared with those from ELISAs using protease negative BmNPV displaying beta1,3 N-acetylglucosaminyltransferase 2 fused with the gene encoding GFPuv (GGT2) (BmNPV-CP--GGT2) particles, which do not display hPRR on their surfaces. CONCLUSION: The display of on the surface of the BmNPV particles will be useful for the detection of protein-protein interactions and the screening of inhibitors and drugs in their roles as nanobioparticles. PMID- 21635721 TI - Researchers' experience with project management in health and medical research: results from a post-project review. AB - BACKGROUND: Project management is widely used to deliver projects on time, within budget and of defined quality. However, there is little published information describing its use in managing health and medical research projects. We used project management in the Alcohol and Pregnancy Project (2006-2008) http://www.ichr.uwa.edu.au/alcoholandpregnancy and in this paper report researchers' opinions on project management and whether it made a difference to the project. METHODS: A national interdisciplinary group of 20 researchers, one of whom was the project manager, formed the Steering Committee for the project. We used project management to ensure project outputs and outcomes were achieved and all aspects of the project were planned, implemented, monitored and controlled. Sixteen of the researchers were asked to complete a self administered questionnaire for a post-project review. RESULTS: The project was delivered according to the project protocol within the allocated budget and time frame. Fifteen researchers (93.8%) completed a questionnaire. They reported that project management increased the effectiveness of the project, communication, teamwork, and application of the interdisciplinary group of researchers' expertise. They would recommend this type of project management for future projects. CONCLUSIONS: Our post-project review showed that researchers comprehensively endorsed project management in the Alcohol and Pregnancy Project and agreed that project management had contributed substantially to the research. In future, we will project manage new projects and conduct post-project reviews. The results will be used to encourage continuous learning and continuous improvement of project management, and provide greater transparency and accountability of health and medical research. The use of project management can benefit both management and scientific outcomes of health and medical research projects. PMID- 21635723 TI - Abdominal pain with a twist. AB - Malrotation in children is due to either an incomplete or non-rotation of the foetal mid-gut during perinatal development. Presentation is usually in the first few weeks of life, often with life-threatening volvulus and ischaemia. However, it can be a rare cause of abdominal pain in older children and young adults. We present such a case, as a reminder to emergency physicians that malrotation should be considered in the differential diagnosis of recurrent or chronic abdominal pain not only in children but also in adolescents. PMID- 21635722 TI - Prevalence and associated factors of depressive and anxiety symptoms during pregnancy: a population based study in rural Bangladesh. AB - BACKGROUND: Few studies have examined the associated factors of antepartum depressive and anxiety symptoms (ADS and AAS) in low-income countries, yet the World Health Organization identifies depressive disorders as the second leading cause of global disease burden by 2020. There is a paucity of research on mental disorders and their predictors among pregnant women in Bangladesh. This study aims to estimate the prevalence of depressive and anxiety symptoms and explore the associated factors in a cross-section of rural Bangladeshi pregnant women. METHODS: The study used cross-sectional data originating from a rural community based prospective cohort study of 720 randomly selected women in their third trimester of pregnancy from a district of Bangladesh. The validated Bangla version of the Edinburgh Postnatal Depression Scale was used to measure ADS, and a trait anxiety inventory to assess general anxiety symptoms. Background information was collected using a structured questionnaire at the respondents' homes. RESULTS: Prevalence of ADS was 18% and AAS 29%. Women's literacy (OR 0.59, 95% CI 0.37-0.95), poor partner relationship (OR 2.23, 95% CI 3.37-3.62), forced sex (OR 1.95, 95% CI 1.01-3.75), physical violence by spouse (OR 1.69, 95% CI 1.02-2.80), and previous depression (OR 4.62 95% CI 2.72-7.85) were found to be associated with ADS. The associated factors of AAS were illiteracy, poor household economy, lack of practical support, physical partner violence, violence during pregnancy, and interaction between poor household economy and poor partner relationship. CONCLUSION: Depressive and anxiety symptoms are found to occur commonly during pregnancy in Bangladesh, drawing attention to a need to screen for depression and anxiety during antenatal care. Policies aimed at encouraging practical support during pregnancy, reducing gender-based violence, supporting women with poor partner relationships, and identifying previous depression may ameliorate the potentially harmful consequences of antepartum depression and anxiety for the women and their family, particularly children. PMID- 21635724 TI - Etiological spectrum, injury characteristics and treatment outcome of maxillofacial injuries in a Tanzanian teaching hospital. AB - BACKGROUND: Maxillofacial injuries pose a therapeutic challenges to trauma, maxillofacial and plastic surgeons practicing in developing countries. This study was carried out to determine the etiology, injury characteristics and management outcome of maxillofacial injuries at our teaching hospital. PATIENTS AND METHODS: A prospective hospital based study of maxillofacial injury patients was carried out at Bugando Medical Centre from November 2008 to October 2009. Data was collected using a structured questionnaire and analyzed using SPPS computer software version 11.5. RESULTS: A total of 154 patients were studied. Males outnumbered females by a ratio of 2.7:1. Their mean age was 28.32 +/- 16.48 years and the modal age group was 21-30 years. Most injuries were caused by road traffic crushes (57.1%), followed by assault and falls in 16.2% and 14.3% respectively. Soft tissue injuries and mandibular fractures were the most common type of injuries. Head/neck (53.1%) and limb injuries (28.1%) were the most prevalent associated injuries. Surgical debridement (95.1%) was the most common surgical procedures. Closed reduction of maxillofacial fractures was employed in 81.5% of patients. Open reduction and internal fixation was performed in 6.8% of cases. Complications occurred in 24% of patients, mainly due to infection and malocclusion. The mean duration of hospital stay was 18.12 +/- 12.24 days. Mortality rate was 11.7%. CONCLUSION: Road traffic crashes remain the major etiological factor of maxillofacial injuries in our setting. Measures on prevention of road traffic crashes should be strongly emphasized in order to reduce the occurrence of these injuries. PMID- 21635725 TI - Antibodies on demand: a fast method for the production of human scFvs with minimal amounts of antigen. AB - BACKGROUND: Antibodies constitute a powerful tool to study protein function, protein localization and protein-protein interactions, as well as for diagnostic and therapeutic purposes. High-throughput antibody development requires faster methodologies with lower antigen consumption. RESULTS: Here, we describe a novel methodology to select human monoclonal recombinant antibodies by combining in vitro protein expression, phage display antibody libraries and antibody microarrays. The application of this combination of methodologies permitted us to generate human single-chain variable fragments (scFvs) against two proteins: green fluorescent protein (GFP) and thioredoxin (Trx) in a short time, using as low as 5 MUg of purified protein. These scFvs showed specific reactivity against their respective targets and worked well by ELISA and western blot. The scFvs were able to recognise as low as 31 ng of protein of their respective targets by western blot. CONCLUSION: This work describes a novel and miniaturized methodology to obtain human monoclonal recombinant antibodies against any target in a shorter time than other methodologies using only 5 MUg of protein. The protocol could be easily adapted to a high-throughput procedure for antibody production. PMID- 21635726 TI - A survey of the transmission of infectious diseases/infections between wild and domestic ungulates in Europe. AB - The domestic animals/wildlife interface is becoming a global issue of growing interest. However, despite studies on wildlife diseases being in expansion, the epidemiological role of wild animals in the transmission of infectious diseases remains unclear most of the time. Multiple diseases affecting livestock have already been identified in wildlife, especially in wild ungulates. The first objective of this paper was to establish a list of infections already reported in European wild ungulates. For each disease/infection, three additional materials develop examples already published, specifying the epidemiological role of the species as assigned by the authors. Furthermore, risk factors associated with interactions between wild and domestic animals and regarding emerging infectious diseases are summarized. Finally, the wildlife surveillance measures implemented in different European countries are presented. New research areas are proposed in order to provide efficient tools to prevent the transmission of diseases between wild ungulates and livestock. PMID- 21635727 TI - Characterisation of the transcriptome of a wild great tit Parus major population by next generation sequencing. AB - BACKGROUND: The recent development of next generation sequencing technologies has made it possible to generate very large amounts of sequence data in species with little or no genome information. Combined with the large phenotypic databases available for wild and non-model species, these data will provide an unprecedented opportunity to "genomicise" ecological model organisms and establish the genetic basis of quantitative traits in natural populations. RESULTS: This paper describes the sequencing, de novo assembly and analysis from the transcriptome of eight tissues of ten wild great tits. Approximately 4.6 million sequences and 1.4 billion bases of DNA were generated and assembled into 95,979 contigs, one third of which aligned with known Taeniopygia guttata (zebra finch) and Gallus gallus (chicken) transcripts. The majority (78%) of the remaining contigs aligned within or very close to regions of the zebra finch genome containing known genes, suggesting that they represented precursor mRNA rather than untranscribed genomic DNA. More than 35,000 single nucleotide polymorphisms and 10,000 microsatellite repeats were identified. Eleven percent of contigs were expressed in every tissue, while twenty one percent of contigs were expressed in only one tissue. The function of those contigs with strong evidence for tissue specific expression and contigs expressed in every tissue was inferred from the gene ontology (GO) terms associated with these contigs; heart and pancreas had the highest number of highly tissue specific GO terms (21.4% and 28.5% respectively). CONCLUSIONS: In summary, the transcriptomic data generated in this study will contribute towards efforts to assemble and annotate the great tit genome, as well as providing the markers required to perform gene mapping studies in wild populations. PMID- 21635728 TI - Neorickettsia risticii surface-exposed proteins: proteomics identification, recognition by naturally-infected horses, and strain variations. AB - Neorickettsia risticii is the Gram-negative, obligate, and intracellular bacterial pathogen responsible for Potomac horse fever (PHF): an important acute systemic disease of horses. N. risticii surface proteins, critical for immune recognition, have not been thoroughly characterized. In this paper, we identified the 51-kDa antigen (P51) as a major surface-exposed outer membrane protein of older and contemporary strains of N. risticii through mass spectrometry of streptavidin-purified biotinylated surface-labeled proteins. Western blot analysis of sera from naturally-infected horses demonstrated universal and strong recognition of recombinant P51 over other Neorickettsia recombinant proteins. Comparisons of amino acid sequences for predicted secondary structures of P51, as well as Neorickettsia surface proteins 2 (Nsp2) and 3 (Nsp3) among N. risticii strains from horses with PHF during a 26-year period throughout the United States revealed that the majority of variations among strains were concentrated in regions predicted to be external loops of their beta-barrel structures. Large insertions or deletions occurred within a tandem-repeat region in Ssa3. These data demonstrate patterns of geographical association for P51 and temporal associations for Nsp2, Nsp3, and Ssa3, indicating evolutionary trends for these Neorickettsia surface antigen genes. This study showed N. risticii surface protein population dynamics, providing groundwork for designing immunodiagnostic targets for PHF. PMID- 21635729 TI - Characterization of porcine dendritic cell response to Streptococcus suis. AB - Streptococcus suis is a major swine pathogen and important zoonotic agent causing mainly septicemia and meningitis. However, the mechanisms involved in host innate and adaptive immune responses toward S. suis as well as the mechanisms used by S. suis to subvert these responses are unknown. Here, and for the first time, the ability of S. suis to interact with bone marrow-derived swine dendritic cells (DCs) was evaluated. In addition, the role of S. suis capsular polysaccharide in modulation of DC functions was also assessed. Well encapsulated S. suis was relatively resistant to phagocytosis, but it increased the relative expression of Toll-like receptors 2 and 6 and triggered the release of several cytokines by DCs, including IL-1beta, IL-6, IL-8, IL-12p40 and TNF-alpha. The capsular polysaccharide was shown to interfere with DC phagocytosis; however, once internalized, S. suis was readily destroyed by DCs independently of the presence of the capsular polysaccharide. Cell wall components were mainly responsible for DC activation, since the capsular polysaccharide-negative mutant induced higher cytokine levels than the wild-type strain. The capsular polysaccharide also interfered with the expression of the co-stimulatory molecules CD80/86 and MHC-II on DCs. To conclude, our results show for the first time that S. suis interacts with swine origin DCs and suggest that these cells might play a role in the development of host innate and adaptive immunity during an infection with S. suis serotype 2. PMID- 21635730 TI - Thrombospondin 1 in hypoxia-conditioned media blocks the growth of human microvascular endothelial cells and is increased in systemic sclerosis tissues. AB - BACKGROUND: Systemic sclerosis (SSc) is a chronic inflammatory autoimmune disease characterised by vascular dysfunction and damage, excess collagen deposition and subsequent organ manifestations. Vasculopathy is an early feature of the disease which leads to a chronic hypoxic environment in the tissues. Paradoxically, there is a lack of angiogenesis. We hypothesised that this may in part be due to a nonphysiological, overriding upregulation in antiangiogenic factors produced by the hypoxic tissues. We considered thrombospondin 1 (TSP-1) as a candidate antiangiogenic factor. RESULTS: Conditioned media from human microvascular endothelial cells cultured in both normoxic and hypoxic environments were able to block endothelial cell proliferation, with the latter environment having a more profound effect. Filtration to remove > 100-kDa proteins or heparin-binding proteins from the conditioned media eliminated their antiproliferative effect. TSP-1 was expressed in high concentrations in the hypoxic media, as was vascular endothelial growth factor (VEGF). Depletion of TSP-1 from the media by immunoprecipitation reduced the antiproliferative effect. We then show that, in a dose-dependent fashion, recombinant TSP-1 blocks the proliferation of endothelial cells. Immunohistochemistry of skin biopsy material revealed that TSP-1 expression was significantly higher throughout the skin of patients with SSc compared with healthy controls. CONCLUSIONS: Despite the environment of chronic tissue hypoxia in SSc, there is a paradoxical absence of angiogenesis. This is thought to be due in part to aberrant expression of antiangiogenic factors, including TSP-1. We have demonstrated that TSP-1 is released in high concentrations by hypoxic endothelial cells. The conditioned media from these cells is able to block proliferation and induce apoptosis in microvascular endothelial cells, an effect that is reduced when TSP-1 is immunoprecipitated out. Further, we have shown that recombinant TSP-1 is able to block proliferation and induce apoptosis at concentrations consistent with those found in the plasma of patients with SSc and that its effect occurs in the presence of elevated VEGF levels. Taken together, these data are consistent with a model wherein injured microvascular cells in SSc fail to repair because of dysregulated induction of TSP-1 in the hypoxic tissues. PMID- 21635731 TI - Ostreid herpesvirus 1 detection and relationship with Crassostrea gigas spat mortality in France between 1998 and 2006. AB - Since its molecular characterisation, Ostreid herpesvirus 1 (OsHV-1) has been regularly detected in Crassostrea gigas in France. Although its pathogenicity was demonstrated on larval stages, its involvement during mortality outbreaks at the juvenile stage was highly suspected but not evidenced. To investigate mortality outbreaks, the French National Network for Surveillance and Monitoring of Mollusc Health (REPAMO) carried out two surveys in juvenile C. gigas. The first survey lasted from 1998 to 2006 and was an epidemiological inquiry occurring when oyster farmers reported mortality outbreaks. The second survey, a longitudinal one, was set up in 1998 to complete the network observations on OsHV-1. Data analysis showed a specific pattern of mortality outbreaks associated with OsHV-1 detection. Ostreid herpesvirus 1 detection mainly appeared during the summer, suggesting the influence of the seawater temperature on its occurrence. It mostly presented a patchy distribution in the field in contrast to the nursery. Significant relationship between OsHV-1 detection and spat mortality was found, preferentially in sheltered and closed environments. The longitudinal survey confirmed most of the network observations. Although subsequent works particularly epidemiological surveys would be useful to confirm the causal link between the detection of OsHV-1 and the mortality outbreaks in juvenile C. gigas, the role of OsHV-1 in oyster mortality is progressing. PMID- 21635732 TI - A single vaccination of commercial broilers does not reduce transmission of H5N1 highly pathogenic avian influenza. AB - Vaccination of chickens has become routine practice in Asian countries in which H5N1 highly pathogenic avian influenza (HPAI) is endemically present. This mainly applies to layer and breeder flocks, but broilers are usually left unvaccinated. Here we investigate whether vaccination is able to reduce HPAI H5N1 virus transmission among broiler chickens. Four sets of experiments were carried out, each consisting of 22 replicate trials containing a pair of birds. Experiments 1 3 were carried out with four-week-old birds that were unvaccinated, and vaccinated at day 1 or at day 10 of age. Experiment 4 was carried out with unvaccinated day-old broiler chicks. One chicken in each trial was inoculated with H5N1 HPAI virus. One chicken in each trial was inoculated with virus. The course of the infection chain was monitored by serological analysis, and by virus isolation performed on tracheal and cloacal swabs. The analyses were based on a stochastic SEIR model using a Bayesian inferential framework. When inoculation was carried out at the 28th day of life, transmission was efficient in unvaccinated birds, and in birds vaccinated at first or tenth day of life. In these experiments estimates of the latent period (~1.0 day), infectious period (~3.3 days), and transmission rate parameter (~1.4 per day) were similar, as were estimates of the reproduction number (~4) and generation interval (~1.4 day). Transmission was significantly less efficient in unvaccinated chickens when inoculation was carried out on the first day of life. These results show that vaccination of broiler chickens does not reduce transmission, and suggest that this may be due to the interference of maternal immunity. PMID- 21635734 TI - The prevalence of polypharmacy in elderly attenders to an emergency department - a problem with a need for an effective solution. AB - We studied the prevalence of polypharmacy in attenders aged 75 years and over to an emergency department (ED) in North London over a period of 1 month. We identified 467 patients in this age group. Analysis of medications being prescribed revealed at least 82 patients on medication with the potential for adverse interaction. There is a need for ED-initiated strategies to identify interactions and for pathways to allow for medication review. PMID- 21635733 TI - Extensive production of Neospora caninum tissue cysts in a carnivorous marsupial succumbing to experimental neosporosis. AB - Experimental infections of Sminthopsis crassicaudata, the fat-tailed dunnart, a carnivorous marsupial widely distributed throughout the arid and semi-arid zones of Australia, show that this species can act as an intermediate host for Neospora caninum. In contrast to existing models that develop relatively few N. caninum tissue cysts, dunnarts offer a new animal model in which active neosporosis is dominated by tissue cyst production. The results provide evidence for a sylvatic life cycle of N. caninum in Australia between marsupials and wild dogs. It establishes the foundation for an investigation of the impact and costs of neosporosis to wildlife. PMID- 21635735 TI - Differential properties of human ACL and MCL stem cells may be responsible for their differential healing capacity. AB - BACKGROUND: The human anterior cruciate ligament (hACL) and medial collateral ligament (hMCL) of the knee joint are frequently injured, especially in athletic settings. It has been known that, while injuries to the MCL typically heal with conservative treatment, ACL injuries usually do not heal. As adult stem cells repair injured tissues through proliferation and differentiation, we hypothesized that the hACL and hMCL contain stem cells exhibiting unique properties that could be responsible for the differential healing capacity of the two ligaments. METHODS: To test the above hypothesis, we derived ligament stem cells from normal hACL and hMCL samples from the same adult donors using tissue culture techniques and characterized their properties using immunocytochemistry, RT-PCR, and flow cytometry. RESULTS: We found that both hACL stem cells (hACL-SCs) and hMCL stem cells (hMCL-SCs) formed colonies in culture and expressed stem cell markers nucleostemin and stage-specific embryonic antigen-4 (SSEA-4). Moreover, both hACL SCs and hMCL-SCs expressed CD surface markers for mesenchymal stem cells, including CD44 and CD90, but not those markers for vascular cells, CD31, CD34, CD45, and CD146. However, hACL-SCs differed from hMCL-SCs in that the size and number of hACL-SC colonies in culture were much smaller and grew more slowly than hMCL-SC colonies. Moreover, fewer hACL-SCs in cell colonies expressed stem cell markers STRO-1 and octamer-binding transcription factor-4 (Oct-4) than hMCL-SCs. Finally, hACL-SCs had less multi-differentiation potential than hMCL-SCs, evidenced by differing extents of adipogenesis, chondrogenesis, and osteogenesis in the respective induction media. CONCLUSIONS: This study shows for the first time that hACL-SCs are intrinsically different from hMCL-SCs. We suggest that the differences in their properties contribute to the known disparity in healing capabilities between the two ligaments. PMID- 21635736 TI - Escape is a more common mechanism than avidity reduction for evasion of CD8+ T cell responses in primary human immunodeficiency virus type 1 infection. AB - BACKGROUND: CD8+ T cells play an important role in control of viral replication during acute and early human immunodeficiency virus type 1 (HIV-1) infection, contributing to containment of the acute viral burst and establishment of the prognostically-important persisting viral load. Understanding mechanisms that impair CD8+ T cell-mediated control of HIV replication in primary infection is thus of importance. This study addressed the relative extent to which HIV specific T cell responses are impacted by viral mutational escape versus reduction in response avidity during the first year of infection. RESULTS: 18 patients presenting with symptomatic primary HIV-1 infection, most of whom subsequently established moderate-high persisting viral loads, were studied. HIV specific T cell responses were mapped in each individual and responses to a subset of optimally-defined CD8+ T cell epitopes were followed from acute infection onwards to determine whether they were escaped or declined in avidity over time. During the first year of infection, sequence variation occurred in/around 26/33 epitopes studied (79%). In 82% of cases of intra-epitopic sequence variation, the mutation was confirmed to confer escape, although T cell responses were subsequently expanded to variant sequences in some cases. In contrast, < 10% of responses to index sequence epitopes declined in functional avidity over the same time-frame, and a similar proportion of responses actually exhibited an increase in functional avidity during this period. CONCLUSIONS: Escape appears to constitute a much more important means of viral evasion of CD8+ T cell responses in acute and early HIV infection than decline in functional avidity of epitope-specific T cells. These findings support the design of vaccines to elicit T cell responses that are difficult for the virus to escape. PMID- 21635737 TI - Conformational alterations in the CD4 binding cavity of HIV-1 gp120 influencing gp120-CD4 interactions and fusogenicity of HIV-1 envelopes derived from brain and other tissues. AB - BACKGROUND: CD4-binding site (CD4bs) alterations in gp120 contribute to HIV-1 envelope (Env) mediated fusogenicity and the ability of gp120 to utilize low levels of cell-surface CD4. In a recent study, we constructed three-dimensional models of gp120 to illustrate CD4bs conformations associated with enhanced fusogenicity and enhanced CD4-usage of a modestly-sized panel of blood-derived HIV-1 Envs (n = 16). These conformations were characterized by a wider aperture of the CD4bs cavity, as constrained by the inner-most atoms at the gp120 V1V2 stem and the V5 loop. Here, we sought to provide further validation of the utility of these models for understanding mechanisms that influence Env function, by characterizing the structure-function relationships of a larger panel of Envs derived from brain and other tissues (n = 81). FINDINGS: Three-dimensional models of gp120 were generated by our recently validated homology modelling protocol. Analysis of predicted CD4bs structures showed correlations between the aperture width of the CD4bs cavity and ability of the Envs to mediate cell-cell fusion, scavenge low-levels of cell-surface CD4, bind directly to soluble CD4, and bind to the Env mAb IgG1b12 whose epitope overlaps the gp120 CD4bs. These structural alterations in the CD4bs cavity were associated with repositioning of the V5 loop. CONCLUSIONS: Using a large, independent panel of Envs, we can confirm the utility of three-dimensional gp120 structural models for illustrating CD4bs alterations that can affect Env function. Furthermore, we now provide new evidence that these CD4bs alterations augment the ability of gp120 to interact with CD4 by increasing the exposure of the CD4bs. PMID- 21635739 TI - A new cultivation method for microbial oil production: cell pelletization and lipid accumulation by Mucor circinelloides. AB - The recent energy crisis has triggered significant attention on the microbial synthesis of lipids, which comprise the raw material for biodiesel production. Microbial oil accumulation with filamentous fungi has great potential because filamentous fungi can form pellets during cell growth, and these pellets are much easier to harvest from cell broth. This paper focuses on the cell pelletization process of the oleaginous Mucor circinelloides. We have studied the effect of various cultural conditions on pelletized cell growth and lipid accumulation. This study is the first to report that pH adjustment during cell growth plays a key role in pellet formation of M. circinelloides and describes a handy method by which to induce cell pelletization in submerged fungal cultivation. Our study reveals that cell growth and lipid production are not significantly affected by pelletization and that lipid accumulation is triggered at stressed conditions, such as a high carbon-to-nitrogen ratio and high temperature. PMID- 21635738 TI - Appraisal of literature reviews on end-of-life care for minority ethnic groups in the UK and a critical comparison with policy recommendations from the UK end-of life care strategy. AB - BACKGROUND: Evidence of low end-of-life (EoL) care service use by minority ethnic groups in the UK has given rise to a body of research and a number of reviews of the literature. This article aims to review and evaluate literature reviews on minority ethnic groups and EoL care in the UK and assess their suitability as an evidence base for policy. METHODS: Systematic review. Searches were carried out in thirteen electronic databases, eight journals, reference lists, and grey literature. Reviews were included if they concerned minority ethnic groups and EoL care in the UK. Reviews were graded for quality and key themes identified. RESULTS: Thirteen reviews (2001-2009) met inclusion criteria. Seven took a systematic approach, of which four scored highly for methodological quality (a mean score of six, median seven). The majority of systematic reviews were therefore of a reasonable methodological quality. Most reviews were restricted by ethnic group, aspect of EoL care, or were broader reviews which reported relevant findings. Six key themes were identified. CONCLUSIONS: A number of reviews were systematic and scored highly for methodological quality. These reviews provide a good reflection of the primary evidence and could be used to inform policy. The complexity and inter-relatedness of factors leading to low service use was recognised and reflected in reviews' recommendations for service improvement. Recommendations made in the UK End-of-Life Care Strategy were limited in comparison, and the Strategy's evidence base concerning minority ethnic groups was found to be narrow. Future policy should be embedded strongly in the evidence base to reflect the current literature and minimise bias. PMID- 21635740 TI - Concurrent detection of autolysosome formation and lysosomal degradation by flow cytometry in a high-content screen for inducers of autophagy. AB - BACKGROUND: Autophagy mediates lysosomal degradation of cytosolic components. Recent work has associated autophagic dysfunction with pathologies, including cancer and cardiovascular disease. To date, the identification of clinically applicable drugs that modulate autophagy has been hampered by the lack of standardized assays capable of precisely reporting autophagic activity. RESULTS: We developed and implemented a high-content, flow-cytometry-based screening approach for rapid, precise, and quantitative measurements of pharmaceutical control over autophagy. Our assay allowed for time-resolved individual measurements of autolysosome formation and degradation, and endolysosomal activities under both basal and activated autophagy conditions. As proof of concept, we analyzed conventional autophagy regulators, including cardioprotective compounds aminoimidazole carboxamide ribonucleotide (AICAR), rapamycin, and resveratrol, and revealed striking conditional dependencies of rapamycin and autophagy inhibitor 3-methyladenine (3-MA). To identify novel autophagy modulators with translational potential, we screened the Prestwick Chemical Library of 1,120 US Food and Drug Administration (FDA)-approved compounds for impact on autolysosome formation. In all, 38 compounds were identified as potential activators, and 36 as potential inhibitors of autophagy. Notably, amongst the autophagy enhancers were cardiac glycosides, from which we selected digoxin, strophanthidin, and digoxigenin for validation by standard biochemical and imaging techniques. We report the induction of autophagic flux by these cardiac glycosides, and the concentrations allowing for specific enhancement of autophagic activities without impact on endolysosomal activities. CONCLUSIONS: Our systematic analysis of autophagic and endolysosomal activities outperformed conventional autophagy assays and highlights the complexity of drug influence on autophagy. We demonstrate conditional dependencies of established regulators. Moreover, we identified new autophagy regulators and characterized cardiac glycosides as novel potent inducers of autophagic flux. PMID- 21635741 TI - Cluster analysis of behavioural and event-related potentials during a contingent negative variation paradigm in remitting-relapsing and benign forms of multiple sclerosis. AB - BACKGROUND: Event-related potentials (ERPs) may be used as a highly sensitive way of detecting subtle degrees of cognitive dysfunction. On the other hand, impairment of cognitive skills is increasingly recognised as a hallmark of patients suffering from multiple sclerosis (MS). We sought to determine the psychophysiological pattern of information processing among MS patients with the relapsing-remitting form of the disease and low physical disability considered as two subtypes: 'typical relapsing-remitting' (RRMS) and 'benign MS' (BMS). Furthermore, we subjected our data to a cluster analysis to determine whether MS patients and healthy controls could be differentiated in terms of their psychophysiological profile. METHODS: We investigated MS patients with RRMS and BMS subtypes using event-related potentials (ERPs) acquired in the context of a Posner visual-spatial cueing paradigm. Specifically, our study aimed to assess ERP brain activity in response preparation (contingent negative variation -CNV) and stimuli processing in MS patients. Latency and amplitude of different ERP components (P1, eN1, N1, P2, N2, P3 and late negativity -LN) as well as behavioural responses (reaction time -RT; correct responses -CRs; and number of errors) were analyzed and then subjected to cluster analysis. RESULTS: Both MS groups showed delayed behavioural responses and enhanced latency for long-latency ERP components (P2, N2, P3) as well as relatively preserved ERP amplitude, but BMS patients obtained more important performance deficits (lower CRs and higher RTs) and abnormalities related to the latency (N1, P3) and amplitude of ERPs (eCNV, eN1, LN). However, RRMS patients also demonstrated abnormally high amplitudes related to the preparation performance period of CNV (cCNV) and post processing phase (LN). Cluster analyses revealed that RRMS patients appear to make up a relatively homogeneous group with moderate deficits mainly related to ERP latencies, whereas BMS patients appear to make up a rather more heterogeneous group with more severe information processing and attentional deficits. CONCLUSIONS: Our findings are suggestive of a slowing of information processing for MS patients that may be a consequence of demyelination and axonal degeneration, which also seems to occur in MS patients that show little or no progression in the physical severity of the disease over time. PMID- 21635742 TI - Process of distant lymph node metastasis in colorectal carcinoma: implication of extracapsular invasion of lymph node metastasis. AB - BACKGROUND: We previously demonstrated that extracapsular invasion (ECI) at a metastatic sentinel node was significantly associated with the presence of positive non-sentinel nodes in patients with breast cancer. However, the mechanism of metastatic spreading of tumor cells to distant lymph nodes in patients with colorectal carcinoma is not fully understood. In this study, we investigated the factors that may determine the likelihood of additional regional lymph node metastasis when metastasis is found in nodes at the N1 site in colorectal cancer, especially focusing on the presence of ECI. METHODS: Two hundred and twenty-eight consecutive patients who underwent colorectal resection were identified for inclusion in this study, of which 37 (16.2%) had positive lymph nodes at the N1 site. Six of these 37 cases had additional metastasis in N2 site lymph nodes. We reviewed the clinicopathological features of these cases and performed statistical analysis of the data. RESULTS: In the univariate analysis ECI at the N1 site was the only factor significantly associated with the presence of cancer cells in the N2 site. Other factors, including number of positive lymph nodes, lymphovascular invasion of the primary tumor, tumor size and tumor depth of invasion, were not associated with metastatic involvement at the N2 site. CONCLUSIONS: Our results suggest that the presence of ECI at metastatic lymph nodes at the N1 site is correlated with further metastasis at the N2 site. These findings imply the possibility that ECI might indicate the ability of colorectal tumor cells to disseminate to distant lymph nodes. PMID- 21635743 TI - Retraction. MicroRNA expression profiling during the life cycle of the silkworm (Bombyx mori). PMID- 21635744 TI - Non medical prescribing leads views on their role and the implementation of non medical prescribing from a multi-organisational perspective. AB - BACKGROUND: In the United Kingdom, non-medical prescribing (NMP) has been identified as one way to improve healthcare quality and efficiency. Healthcare organisations are charged with overseeing the clinical governance of NMP and guidance recommends the identification of a lead director to be responsible for its implementation. While over twelve million items are prescribed each year by the 50,000 qualified NMPs its uptake is inconsistent. Several studies have explored the barriers to NMP at a practice level, however little is known about the role the NMP lead and the implementation of NMP from an organisational perspective. The aim of this research was to explore the role of the organisational NMP lead across a range of practice settings within one Strategic Health Authority (SHA) and consider the development of NMP from a multi organisational perspective. METHODS: Semi-structured telephone interviews with 28 NMP leads across one SHA were undertaken by a trained qualitative researcher. Interviews addressed the purpose of the role and difficulties encountered; audiotapes were transcribed, coded and themes were identified. RESULTS: The NMP lead role comprised of four main functions; communication, coordinating, clinical governance and support. Factors hampering progress in overseeing the safe development of NMP included lack of clarity about the NMP lead role and responsibilities, strategic support and a lack of protected time. The extent to which clinical governance systems were in place across organisations was inconsistent. Where a strategic approach to its development was adopted, fewer barriers were encountered and NMP was more likely to become embedded within organisations. CONCLUSIONS: The significant contribution that NMP leads play in embedding NMP within organisations should be acknowledged by clearer national guidance for this role and its responsibilities. Greater standardisation and consistency is required of clinical governance systems if quality and safety is to be ensured given the expanding development of NMP. The extent to which NMP is in place worldwide differs. However, our findings will be of interest to policymakers in other countries involved in the development and implementation of this role. PMID- 21635745 TI - Distinct different expression of Th17 and Th9 cells in coxsackie virus B3-induced mice viral myocarditis. AB - BACKGROUND: Recently, a new subset of CD4+T helper(Th) cell that predominantly secret cytokine interleukin-9(IL-9) is identified, termed Th9 cell. It has been reported to participate in tissue inflammation and autoimmune responses, and induce disease which differed from Th17 cells. Th17 cells have been shown to play a critical role in viral myocarditis (VMC), but whether Th9 cells are involved in the pathogenesis of VMC remains unclear. RESULTS: BALB/c mice were intraperitoneally (i.p) injected with coxsackie virus B3(CVB3) for establishing VMC models. Control mice were treated with phosphate-buffered saline i.p. On day 0,7,14,21,28,35,42 after injection, myocardial histopathological changes were evaluated by hematoxylin-eosin staining. Splenic Th17 and Th9 cells subsets were analyzed by flow cytometry. And cardiac IL-17, IL-9 mRNA were measured by semi quantitative reverse transcription-PCR and nested PCR, respectively. Results showed the levels of Th17 cells and IL-17 mRNA obviously increased in VMC mice on 7 day after infection, peaked on day 28, and highly persisted to at least day 42 (p < 0.05). While the frequencies of Th9 cells and IL-9 mRNA showed no significant difference between VMC and control group throughout the course of the experiment(p > 0.05). CONCLUSIONS: It was differentiated Th17 but not Th9 cells significantly elevated in the development of CVB3-induced VMC. The microenvironment of VMC seemed to contribute to the differentiation and proliferation of Th17 rather than Th9 cells. Our preliminary data implied Th9 cells could not protect against VMC nor promote the disease. PMID- 21635746 TI - Systematic review and meta-analysis comparing land and aquatic exercise for people with hip or knee arthritis on function, mobility and other health outcomes. AB - BACKGROUND: Aquatic and land based exercise are frequently prescribed to maintain function for people with arthritis. The relative efficacy of these rehabilitation strategies for this population has not been established.This review investigated the effects of aquatic compared to land based exercise on function, mobility or participants' perception of programs for people with arthritis. METHODS: Medline, CINAHL, AMED and the Cochrane Central Register of Controlled Clinical Trials were searched up to July 2010. Ten randomised, controlled clinical trials that compared land to aquatic exercise for adults with arthritis were included. Study quality was assessed with the PEDro scale. Data relevant to the review question were systematically extracted by two independent reviewers. Standardised mean differences between groups for key outcomes were calculated. Meta-analyses were performed for function, mobility and indices that pooled health outcomes across multiple domains. RESULTS: No differences in outcomes were observed for the two rehabilitation strategies in meta-analysis. There was considerable variability between trials in key program characteristics including prescribed exercises and design quality. Components of exercise programs were poorly reported by the majority of trials. No research was found that examined participant preferences for aquatic compared to land based exercise, identifying this as an area for further research. CONCLUSION: Outcomes following aquatic exercise for adults with arthritis appear comparable to land based exercise. When people are unable to exercise on land, or find land based exercise difficult, aquatic programs provide an enabling alternative strategy. PMID- 21635747 TI - ngs_backbone: a pipeline for read cleaning, mapping and SNP calling using next generation sequence. AB - BACKGROUND: The possibilities offered by next generation sequencing (NGS) platforms are revolutionizing biotechnological laboratories. Moreover, the combination of NGS sequencing and affordable high-throughput genotyping technologies is facilitating the rapid discovery and use of SNPs in non-model species. However, this abundance of sequences and polymorphisms creates new software needs. To fulfill these needs, we have developed a powerful, yet easy-to use application. RESULTS: The ngs_backbone software is a parallel pipeline capable of analyzing Sanger, 454, Illumina and SOLiD (Sequencing by Oligonucleotide Ligation and Detection) sequence reads. Its main supported analyses are: read cleaning, transcriptome assembly and annotation, read mapping and single nucleotide polymorphism (SNP) calling and selection. In order to build a truly useful tool, the software development was paired with a laboratory experiment. All public tomato Sanger EST reads plus 14.2 million Illumina reads were employed to test the tool and predict polymorphism in tomato. The cleaned reads were mapped to the SGN tomato transcriptome obtaining a coverage of 4.2 for Sanger and 8.5 for Illumina. 23,360 single nucleotide variations (SNVs) were predicted. A total of 76 SNVs were experimentally validated, and 85% were found to be real. CONCLUSIONS: ngs_backbone is a new software package capable of analyzing sequences produced by NGS technologies and predicting SNVs with great accuracy. In our tomato example, we created a highly polymorphic collection of SNVs that will be a useful resource for tomato researchers and breeders. The software developed along with its documentation is freely available under the AGPL license and can be downloaded from http://bioinf.comav.upv.es/ngs_backbone/ or http://github.com/JoseBlanca/franklin. PMID- 21635748 TI - Similarities and differences of the soleus and gastrocnemius H-reflexes during varied body postures, foot positions, and muscle function: multifactor designs for repeated measures. AB - BACKGROUND: Although the soleus (Sol), medial gastrocnemius (MG), and lateral gastrocnemius (LG) muscles differ in function, composition, and innervations, it is a common practice is to investigate them as single H-reflex recording. The purpose of this study was to compare H-reflex recordings between these three sections of the triceps surae muscle group of healthy participants while lying and standing during three different ankle positions. METHODS: The Sol, MG and LG muscles' H-reflexes were recorded from ten participants during prone lying and standing with the ankle in neutral, maximum dorsiflexion, and maximum plantarflexion positions. Four traces were averaged for each combination of conditions. Three-way ANOVAs (posture X ankle position X muscle) with planned comparisons were used for statistical comparisons. RESULTS: Although the H-reflex in the three muscle sections differed in latency and amplitude, its dependency on posture and ankle position was similar. The H-reflex amplitudes and maximum H reflex to M-response (H/M) ratios were significantly 1) lower during standing compared to lying with the ankle in neutral, 2) greater during standing with the ankle in plantarflexion compared to neutral, and 3) less with the ankle in dorsiflexion compared to neutral during lying and standing for all muscles (p <= .05). CONCLUSION: Varying demands are required for muscles activated during distinctly different postures and ankle movement tasks. PMID- 21635749 TI - Exploiting MeSH indexing in MEDLINE to generate a data set for word sense disambiguation. AB - BACKGROUND: Evaluation of Word Sense Disambiguation (WSD) methods in the biomedical domain is difficult because the available resources are either too small or too focused on specific types of entities (e.g. diseases or genes). We present a method that can be used to automatically develop a WSD test collection using the Unified Medical Language System (UMLS) Metathesaurus and the manual MeSH indexing of MEDLINE. We demonstrate the use of this method by developing such a data set, called MSH WSD. METHODS: In our method, the Metathesaurus is first screened to identify ambiguous terms whose possible senses consist of two or more MeSH headings. We then use each ambiguous term and its corresponding MeSH heading to extract MEDLINE citations where the term and only one of the MeSH headings co-occur. The term found in the MEDLINE citation is automatically assigned the UMLS CUI linked to the MeSH heading. Each instance has been assigned a UMLS Concept Unique Identifier (CUI). We compare the characteristics of the MSH WSD data set to the previously existing NLM WSD data set. RESULTS: The resulting MSH WSD data set consists of 106 ambiguous abbreviations, 88 ambiguous terms and 9 which are a combination of both, for a total of 203 ambiguous entities. For each ambiguous term/abbreviation, the data set contains a maximum of 100 instances per sense obtained from MEDLINE.We evaluated the reliability of the MSH WSD data set using existing knowledge-based methods and compared their performance to that of the results previously obtained by these algorithms on the pre-existing data set, NLM WSD. We show that the knowledge-based methods achieve different results but keep their relative performance except for the Journal Descriptor Indexing (JDI) method, whose performance is below the other methods. CONCLUSIONS: The MSH WSD data set allows the evaluation of WSD algorithms in the biomedical domain. Compared to previously existing data sets, MSH WSD contains a larger number of biomedical terms/abbreviations and covers the largest set of UMLS Semantic Types. Furthermore, the MSH WSD data set has been generated automatically reusing already existing annotations and, therefore, can be regenerated from subsequent UMLS versions. PMID- 21635750 TI - Using affinity propagation for identifying subspecies among clonal organisms: lessons from M. tuberculosis. AB - BACKGROUND: Classification and naming is a key step in the analysis, understanding and adequate management of living organisms. However, where to set limits between groups can be puzzling especially in clonal organisms. Within the Mycobacterium tuberculosis complex (MTC), the etiological agent of tuberculosis (TB), experts have first identified several groups according to their pattern at repetitive sequences, especially at the CRISPR locus (spoligotyping), and to their epidemiological relevance. Most groups such as "Beijing" found good support when tested with other loci. However, other groups such as T family and T1 subfamily (belonging to the "Euro-American" lineage) correspond to non monophyletic groups and still need to be refined. Here, we propose to use a method called Affinity Propagation that has been successfully used in image categorization to identify relevant patterns at the CRISPR locus in MTC. RESULTS: To adequately infer the relative divergence time between strains, we used a distance method inspired by the recent evolutionary model by Reyes et al. We first confirm that this method performs better than the Jaccard index commonly used to compare spoligotype patterns. Second, we document the support of each spoligotype family among the previous classification using affinity propagation on the international spoligotyping database SpolDB4. This allowed us to propose a consensus assignation for all SpolDB4 spoligotypes. Third, we propose new signatures to subclassify the T family. CONCLUSION: Altogether, this study shows how the new clustering algorithm Affinity Propagation can help building or refining clonal organims classifications. It also describes well-supported families and subfamilies among M. tuberculosis complex, especially inside the modern "Euro-American" lineage. PMID- 21635751 TI - Binding site prediction for protein-protein interactions and novel motif discovery using re-occurring polypeptide sequences. AB - BACKGROUND: While there are many methods for predicting protein-protein interaction, very few can determine the specific site of interaction on each protein. Characterization of the specific sequence regions mediating interaction (binding sites) is crucial for an understanding of cellular pathways. Experimental methods often report false binding sites due to experimental limitations, while computational methods tend to require data which is not available at the proteome-scale. Here we present PIPE-Sites, a novel method of protein specific binding site prediction based on pairs of re-occurring polypeptide sequences, which have been previously shown to accurately predict protein-protein interactions. PIPE-Sites operates at high specificity and requires only the sequences of query proteins and a database of known binary interactions with no binding site data, making it applicable to binding site prediction at the proteome-scale. RESULTS: PIPE-Sites was evaluated using a dataset of 265 yeast and 423 human interacting proteins pairs with experimentally determined binding sites. We found that PIPE-Sites predictions were closer to the confirmed binding site than those of two existing binding site prediction methods based on domain-domain interactions, when applied to the same dataset. Finally, we applied PIPE-Sites to two datasets of 2347 yeast and 14,438 human novel interacting protein pairs predicted to interact with high confidence. An analysis of the predicted interaction sites revealed a number of protein subsequences which are highly re-occurring in binding sites and which may represent novel binding motifs. CONCLUSIONS: PIPE-Sites is an accurate method for predicting protein binding sites and is applicable to the proteome-scale. Thus, PIPE-Sites could be useful for exhaustive analysis of protein binding patterns in whole proteomes as well as discovery of novel binding motifs. PIPE-Sites is available online at http://pipe-sites.cgmlab.org/. PMID- 21635752 TI - Two-tier charging in Maputo Central Hospital: costs, revenues and effects on equity of access to hospital services. AB - BACKGROUND: Special services within public hospitals are becoming increasingly common in low and middle income countries with the stated objective of providing higher comfort services to affluent customers and generating resources for under funded hospitals. In the present study expenditures, outputs and costs are analysed for the Maputo Central Hospital and its Special Clinic with the objective of identifying net resource flows between a system operating two-tier charging, and, ultimately, understanding whether public hospitals can somehow benefit from running Special Clinic operations. METHODS: A combination of step down and bottom-up costing strategies were used to calculate recurrent as well as capital expenses, apportion them to identified cost centres and link costs to selected output measures. RESULTS: The results show that cost differences between main hospital and clinic are marked and significant, with the Special Clinic's cost per patient and cost per outpatient visit respectively over four times and over thirteen times their equivalent in the main hospital. DISCUSSION: While the main hospital cost structure appeared in line with those from similar studies, salary expenditures were found to drive costs in the Special Clinic (73% of total), where capital and drug costs were surprisingly low (2 and 4% respectively). We attributed low capital and drug costs to underestimation by our study owing to difficulties in attributing the use of shared resources and to the Special Clinic's outsourcing policy. The large staff expenditure would be explained by higher physician time commitment, economic rents and subsidies to hospital staff. On the whole it was observed that: (a) the flow of capital and human resources was not fully captured by the financial systems in place and stayed largely unaccounted for; (b) because of the little consideration given to capital costs, the main hospital is more likely to be subsidising its Special Clinic operations, rather than the other way around. CONCLUSION: We conclude that the observed lack of transparency may create scope for an inequitable cross subsidy of private customers by public resources. PMID- 21635753 TI - A randomised controlled trial of an open lung strategy with staircase recruitment, titrated PEEP and targeted low airway pressures in patients with acute respiratory distress syndrome. AB - INTRODUCTION: Tidal volume and plateau pressure minimisation are the standard components of a protective lung ventilation strategy for patients with acute respiratory distress syndrome (ARDS). Open lung strategies, including higher positive end-expiratory pressure (PEEP) and recruitment manoeuvres to date have not proven efficacious. This study examines the effectiveness and safety of a novel open lung strategy, which includes permissive hypercapnia, staircase recruitment manoeuvres (SRM) and low airway pressure with PEEP titration. METHOD: Twenty ARDS patients were randomised to treatment or ARDSnet control ventilation strategies. The treatment group received SRM with decremental PEEP titration and targeted plateau pressure < 30 cm H2O. Gas exchange and lung compliance were measured daily for 7 days and plasma cytokines in the first 24 hours and on days 1, 3, 5 and 7 (mean +/- SE). Duration of ventilation, ICU stay and hospital stay (median and interquartile range) and hospital survival were determined. RESULTS: There were significant overall differences between groups when considering plasma IL-8 and TNF-alpha. For plasma IL-8, the control group was 41% higher than the treatment group over the seven-day period (ratio 1.41 (1.11 to 1.79), P = 0.01), while for TNF-alpha the control group was 20% higher over the seven-day period (ratio 1.20 (1.01 to 1.42) P = 0.05). PaO2/FIO2 (204 +/- 9 versus 165 +/- 9 mmHg, P = 0.005) and static lung compliance (49.1 +/- 2.9 versus 33.7 +/- 2.7 mls/cm H2O, P < 0.001) were higher in the treatment group than the control group over seven days. There was no difference in duration of ventilation (180 (87 to 298) versus 341 (131 to 351) hrs, P = 0.13), duration of ICU stay (9.9 (5.6 to 14.8) versus 16.0 (8.1 to 19.3) days, P = 0.19) and duration of hospital stay (17.9 (13.7 to 34.5) versus 24.7 (20.5 to 39.8) days, P = 0.16) between the treatment and control groups. CONCLUSIONS: This open lung strategy was associated with greater amelioration in some systemic cytokines, improved oxygenation and lung compliance over seven days. A larger trial powered to examine clinically meaningful outcomes is warranted. TRIAL REGISTRATION: ACTRN12607000465459. PMID- 21635754 TI - Outbreak of pneumonia in the setting of fatal pneumococcal meningitis among US Army trainees: potential role of Chlamydia pneumoniae infection. AB - BACKGROUND: Compared to the civilian population, military trainees are often at increased risk for respiratory infections. We investigated an outbreak of radiologically-confirmed pneumonia that was recognized after 2 fatal cases of serotype 7F pneumococcal meningitis were reported in a 303-person military trainee company (Alpha Company). METHODS: We reviewed surveillance data on pneumonia and febrile respiratory illness at the training facility; conducted chart reviews for cases of radiologically-confirmed pneumonia; and administered surveys and collected nasopharyngeal swabs from trainees in the outbreak battalion (Alpha and Hotel Companies), associated training staff, and trainees newly joining the battalion. RESULTS: Among Alpha and Hotel Company trainees, the average weekly attack rates of radiologically-confirmed pneumonia were 1.4% and 1.2% (most other companies at FLW: 0-0.4%). The pneumococcal carriage rate among all Alpha Company trainees was 15% with a predominance of serotypes 7F and 3. Chlamydia pneumoniae was identified from 31% of specimens collected from Alpha Company trainees with respiratory symptoms. CONCLUSION: Although the etiology of the outbreak remains unclear, the identification of both S. pneumoniae and C. pneumoniae among trainees suggests that both pathogens may have contributed either independently or as cofactors to the observed increased incidence of pneumonia in the outbreak battalion and should be considered as possible etiologies in outbreaks of pneumonia in the military population. PMID- 21635755 TI - Distinctions in gastric cancer gene expression signatures derived from laser capture microdissection versus histologic macrodissection. AB - BACKGROUND: Gastric cancer samples obtained by histologic macrodissection contain a relatively high stromal content that may significantly influence gene expression profiles. Differences between the gene expression signature derived from macrodissected gastric cancer samples and the signature obtained from isolated gastric cancer epithelial cells from the same biopsies using laser capture microdissection (LCM) were evaluated for their potential experimental biases. METHODS: RNA was isolated from frozen tissue samples of gastric cancer biopsies from 20 patients using both histologic macrodissection and LCM techniques. RNA from LCM was subject to an additional round of T7 RNA amplification. Expression profiling was performed using Affymetrix HG-U133A arrays. Genes identified in the expression signatures from each tissue processing method were compared to the set of genes contained within chromosomal regions found to harbor copy number aberrations in the tumor samples by array CGH and to proteins previously identified as being overexpressed in gastric cancer. RESULTS: Genes shown to have increased copy number in gastric cancer were also found to be overexpressed in samples obtained by macrodissection (LS P value < 10(-5)), but not in array data generated using microdissection. A set of 58 previously identified genes overexpressed in gastric cancer was also enriched in the gene signature identified by macrodissection (LS P < 10(-5)), but not in the signature identified by microdissection (LS P = 0.013). In contrast, 66 genes previously reported to be underexpressed in gastric cancer were enriched in the gene signature identified by microdissection (LS P < 10-5), but not in the signature identified by macrodissection (LS P = 0.89). CONCLUSIONS: The tumor sampling technique biases the microarray results. LCM may be a more sensitive collection and processing method for the identification of potential tumor suppressor gene candidates in gastric cancer using expression profiling. PMID- 21635756 TI - Retraction: Molecular characterization of Legionella pneumophila-induced interleukin-8 expression in T cells. PMID- 21635757 TI - Perinatal outcomes associated with low birth weight in a historical cohort. AB - OBJECTIVE: To identify perinatal outcomes associated with low birth weight (LBW). METHODS: A retrospective cohort study in a tertiary maternity hospital. Analysis of the database on 43,499 liveborn infants delivered between 1986 and 2004 with low (n = 6,477) and normal (n = 37,467) birth weight. Outcomes associated with LBW were identified through crude and adjusted risk ratio (RR) and 95%CI with bivariate and multivariate analysis. The main outcomes were: onset of labor, mode of delivery, indication for cesarean section; amniotic fluid, fetal heart rate pattern, Apgar score, somatic gestational age, gender and congenital malformation. RESULTS: LBW infants showed more frequently signs of perinatal compromise such as abnormal amniotic fluid volume (especially olygohydramnios), nonreassuring patterns of fetal heart rate, malformation, lower Apgar scores and lower gestational age at birth. They were associated with a greater risk of labor induction and cesarean delivery, but lower risk of forceps. CONCLUSION: There was a clear association between LBW and unfavorable perinatal outcomes. PMID- 21635758 TI - Retraction: NF-kappaB activation by Helicobacter pylori requires Akt-mediated phosphorylation of p65. PMID- 21635759 TI - Interpreting finite element results for brittle materials in endodontic restorations. AB - BACKGROUND: Finite element simulation has been used in last years for analysing the biomechanical performance of post-core restorations in endodontics, but results of these simulations have been interpreted in most of the works using von Mises stress criterion. However, the validity of this failure criterion for brittle materials, which are present in these restorations, is questionable. The objective of the paper is to analyse how finite element results for brittle materials of endodontic restorations should be interpreted to obtain correct conclusions about the possible failure in the restoration. METHODS: Different failure criteria (Von Mises, Rankine, Coulomb-Mohr, Modified Mohr and Christensen) and material strength data (diametral tensile strength and flexural strength) were considered in the study. Three finite element models (FEM) were developed to simulate an endodontic restoration and two typical material tests: diametral tensile test and flexural test. RESULTS: Results showed that the Christensen criterion predicts similar results as the Von Mises criterion for ductile components, while it predicts similar results to all other criteria for brittle components. The different criteria predict different failure points for the diametral tensile test, all of them under multi-axial stress states. All criteria except Von Mises predict failure for flexural test at the same point of the specimen, with this point under uniaxial tensile stress. CONCLUSIONS: From the results it is concluded that the Christensen criterion is recommended for FEM result interpretation in endodontic restorations and that the flexural test is recommended to estimate tensile strength instead of the diametral tensile test. PMID- 21635760 TI - Evaluation of biolistic gene transfer methods in vivo using non-invasive bioluminescent imaging techniques. AB - BACKGROUND: Gene therapy continues to hold great potential for treating many different types of disease and dysfunction. Safe and efficient techniques for gene transfer and expression in vivo are needed to enable gene therapeutic strategies to be effective in patients. Currently, the most commonly used methods employ replication-defective viral vectors for gene transfer, while physical gene transfer methods such as biolistic-mediated ("gene-gun") delivery to target tissues have not been as extensively explored. In the present study, we evaluated the efficacy of biolistic gene transfer techniques in vivo using non-invasive bioluminescent imaging (BLI) methods. RESULTS: Plasmid DNA carrying the firefly luciferase (LUC) reporter gene under the control of the human Cytomegalovirus (CMV) promoter/enhancer was transfected into mouse skin and liver using biolistic methods. The plasmids were coupled to gold microspheres (1 MUm diameter) using different DNA Loading Ratios (DLRs), and "shot" into target tissues using a helium-driven gene gun. The optimal DLR was found to be in the range of 4-10. Bioluminescence was measured using an In Vivo Imaging System (IVIS-50) at various time-points following transfer. Biolistic gene transfer to mouse skin produced peak reporter gene expression one day after transfer. Expression remained detectable through four days, but declined to undetectable levels by six days following gene transfer. Maximum depth of tissue penetration following biolistic transfer to abdominal skin was 200-300 MUm. Similarly, biolistic gene transfer to mouse liver in vivo also produced peak early expression followed by a decline over time. In contrast to skin, however, liver expression of the reporter gene was relatively stable 4-8 days post-biolistic gene transfer, and remained detectable for nearly two weeks. CONCLUSIONS: The use of bioluminescence imaging techniques enabled efficient evaluation of reporter gene expression in vivo. Our results demonstrate that different tissues show different expression kinetics following gene transfer of the same reporter plasmid to different mouse tissues in vivo. We evaluated superficial (skin) and abdominal organ (liver) targets, and found that reporter gene expression peaked within the first two days post transfer in each case, but declined most rapidly in the skin (3-4 days) compared to liver (10-14 days). This information is essential for designing effective gene therapy strategies in different target tissues. PMID- 21635761 TI - Effects of ischemic preconditioning on ischemia/reperfusion-induced arrhythmias by upregulatation of connexin 43 expression. AB - BACKGROUND: The susceptibility of hypertrophied myocardium to ischemia reperfusion injury is associated with increased risk of postoperative arrhythmias. We investigate the effects of ischemic preconditioning (IP) on post ischemic reperfusion arrhythmias in hypertrophic rabbit hearts. METHODS: Thirty three rabbit models of myocardial hypertrophy were randomly divided into three groups of 11 each: non-ischemia-reperfusion group (group A), ischemia-reperfusion group (group B), and ischemic preconditioning group (group C). Another ten healthy rabbits with normal myocardium served as the healthy control group. Rabbit models of myocardial hypertrophy were induced by abdominal aortic banding. Surface electrocardiogram (ECG) was recorded and Curtis-Ravingerova score was used for arrhythmia quantification. Connexin 43 (Cx43) expression was assessed by immunohistochemistry. RESULTS: Ratios of heart weight to body weight and left ventricular weight to body weight increase significantly in the three groups compared with the healthy control group (p < 0.05). Arrhythmia incidence in group C is significantly lower than group B (p < 0.05). Curtis-Ravingerova score in group C is lower than group B (p < 0.05). Cx43 expression area in group A is smaller by comparison with the healthy control group (p < 0.05). Cx43 expression area and fluorescence intensity in group B are reduced by 60.9% and 23.9%, respectively, compared with group A (p < 0.05). In group C, Cx43 expression area increases by 32.5% compared with group B (p < 0.05), and decreases by 54.8% compared with group A (p < 0.05). CONCLUSIONS: The incidence of ischemia/reperfusion-induced arrhythmias in hypertrophic rabbit hearts decreases after IP, which plays an important protecting role on the electrophysiology of hypertrophied myocardium by up-regulating the expression of Cx43. PMID- 21635762 TI - Age differences in heroin and prescription opioid abuse among enrolees into opioid treatment programs. AB - BACKGROUND: In the United States, among those entering opioid treatment programs (OTPs), prescription opioid (PO) abusers tend to be younger than heroin users. Admissions of older persons to OTPs have been increasing, and it is important to understand typical patterns of use among those older enrolees. METHODS: To disentangle the effect of age on recent heroin and PO abuse 29,114 enrolees into 85 OTPs were surveyed across 34 states from 2005-2009. OTPs where PO use was prevalent were oversampled. RESULTS: Mean age was 34; 28% used heroin only. Younger enrolees had increased odds of using POs relative to using heroin only but mixed model analysis showed that much of the total variability in type of use was attributed to variation in age between OTPs rather than within OTPs. CONCLUSIONS: Organizational and cultural phenomena (e.g., OTP characteristics) must be examined to better understand the context of individual characteristics (e.g., age). If nesting of enrolees within OTPs is ignored, then associations that primarily operate at the OTP level may be misinterpreted as exclusively dependent on individuals. PMID- 21635763 TI - A protocol for a systematic review of knowledge translation strategies in the allied health professions. AB - BACKGROUND: Knowledge translation (KT) aims to close the gap between knowledge and practice in order to realize the benefits of research through (a) improved health outcomes, (b) more effective health services and products, and (c) strengthened healthcare systems. While there is some understanding of strategies to put research findings into practice within nursing and medicine, we have limited knowledge of KT strategies in allied health professions. Given the interprofessional nature of healthcare, a lack of guidance for supporting KT strategies in the allied health professions is concerning. Our objective in this study is to systematically review published research on KT strategies in five allied health disciplines. METHODS: A medical research librarian will develop and implement search strategies designed to identify evidence that is relevant to each question of the review. Two reviewers will perform study selection and quality assessment using standard forms. For study selection, data will be extracted by two reviewers. For quality assessment, data will be extracted by one reviewer and verified by a second. Disagreements will be resolved through discussion or third party adjudication. Within each profession, data will be grouped and analyzed by research design and KT strategies using the Effective Practice and Organisation of Care Review Group classification scheme. An overall synthesis across professions will be conducted. SIGNIFICANCE: A uniprofessional approach to KT does not represent the interprofessional context it targets. Our findings will provide the first systematic overview of KT strategies used in allied health professionals' clinical practice, as well as a foundation to inform future KT interventions in allied healthcare settings. PMID- 21635764 TI - Protein kinase C inhibition ameliorates functional endothelial insulin resistance and vascular smooth muscle cell hypersensitivity to insulin in diabetic hypertensive rats. AB - OBJECTIVE: Insulin resistance, diabetes, and hypertension are considered elements of metabolic syndrome which is associated with vascular dysfunction. We investigated whether inhibition of protein kinase C (PKC) would affect vascular function in diabetic hypertensive (DH) rats. METHODS: A combination of type 2 diabetes and arterial hypertension was produced in male Sprague Dawley rats by intrauterine protein deprivation (IUPD) followed by high salt diet. At the age of 32 weeks, DH rats were treated for 2 weeks with the angiotensin-converting enzyme inhibitor captopril (Capto, 30 mg/kg), PKC inhibitor ruboxistaurin (RBX, 50 mg/kg) or vehicle (n = 8 per group) and blood pressure was monitored using telemetry. At the end of experiments, femoral arteries were dissected, and vascular reactivity was evaluated with isovolumic myography. RESULTS: The IUPD followed by high salt diet resulted in significant elevation of plasma glucose, plasma insulin, and blood pressure. Endothelium-dependent vascular relaxation in response to acetylcholine was blunted while vascular contraction in response to phenylephrine was enhanced in the DH rats. Neither Capto nor RBX restored endothelium-dependent vascular relaxation while both suppressed vascular contraction. Ex-vivo incubation of femoral arteries from control rats with insulin induced dose-response vasorelaxation while insulin failed to induce vasorelaxation in the DH rat arteries. In the control arteries treated with endothelial nitric oxide synthase inhibitor L-NAME, insulin induced vasoconstriction that was exacerbated in DH rats. Capto and RBX partially inhibited insulin-stimulated vascular contraction. CONCLUSION: These findings suggest that PKC inhibition ameliorates functional endothelial insulin resistance and smooth muscle cell hypersensitivity to insulin, but does not restore acetylcholine-activated endothelium-dependent vasodilation in DH rats. PMID- 21635765 TI - Premature ventricular contractions originating from the left ventricular septum: results of radiofrequency catheter ablation in twenty patients. AB - BACKGROUND: RFCA has been established as an effective and curative therapy for severely symptomatic PVC from the outflow tract in structurally normal hearts. However, it is unknown whether PVCs originating from the left ventricular septum, are effectively eliminated by RFCA. This study aimed to investigate electrophysiologic characteristics and effects of radiofrequency catheter ablation (RFCA) for patients with symptomatic premature ventricular contraction (PVC) originating from the left ventricular septum without including fascicular PVCs. METHODS: Characteristics of body surface electrocardiogram (ECG) and electrophysiologic recordings endocardiogram in a successful RFCA target were analyzed in 20 patients with symptomatic PVCs originating from the left ventricular septum. RFCA was performed using pace mapping and activation mapping. RESULTS: The QRS morphology of PVCs originating from the left ventricular septum is similar to that seen in fascicular tachycardia. Most of the PVCs originated from the left septum appears in the form of ventricular parasystole. The incidence of ventricular parasystole was 70%. Sustained ventricular tachycardia was not inducible by electrical stimulation and isoproterenol infusion in all 20 patients, ablation at the site recording the earliest Purkinje potential was not effective in all 20 patients, and Purkinje potentials were not identified at successful sites during point mapping. Sixteen patients were successful with RFCA using pace mapping and activation mapping, 3 failed, and 1 recurrent. CONCLUSION: Although the ECG characteristics of the PVCs arising from the left ventricular septum are similar to that seen in fascicular tachycardia, the electrophysiologic characteristics are different between the two types of PVCs. The distinguishing characteristic of the PVCs is that Purkinje potentials were not present at the site of successful ablation, suggesting a myocardial as opposed to fascicular substrate. RFCA is an effective curative therapy for symptomatic PVCs originating from the left ventricular septum (not from the left anterior and posterior fascicle). PMID- 21635766 TI - Ethnobotany of religious and supernatural beliefs of the Mising tribes of Assam with special reference to the 'Dobur Uie'. AB - Assam is very rich in plant biodiversity as well as in ethnic diversity and has a great traditional knowledge base in plant resources. It is inhabited by the largest number of tribes and they lead an intricate life totally dependent on forest plants. The Mising is the major section and second largest tribal community of Assam and have a rich tradition of religion and culture. Their religious practices and beliefs are based on supernaturalism. A study of the plants related to magico religious beliefs in Dobur Uie of Mising is carried out. The results revealed the use of 30 plants belonging to 23 families. All plant species are used both in religious purpose as well as in the treatment of different ailments. Details of the uses of plants and conservational practices employed in Dobur Uie are provided. Our findings on the use of plants in Dobur Uie ritual reflect that some plants are facing problems for survival and they need urgent conservation before their elimination. Because this elimination may threat the rich tradition of Mising culture. Most of the plants that are domesticated for different rituals are almost same in all Mising populated areas. PMID- 21635767 TI - Gene coexpression clusters and putative regulatory elements underlying seed storage reserve accumulation in Arabidopsis. AB - BACKGROUND: In Arabidopsis, a large number of genes involved in the accumulation of seed storage reserves during seed development have been characterized, but the relationship of gene expression and regulation underlying this physiological process remains poorly understood. A more holistic view of this molecular interplay will help in the further study of the regulatory mechanisms controlling seed storage compound accumulation. RESULTS: We identified gene coexpression networks in the transcriptome of developing Arabidopsis (Arabidopsis thaliana) seeds from the globular to mature embryo stages by analyzing publicly accessible microarray datasets. Genes encoding the known enzymes in the fatty acid biosynthesis pathway were found in one coexpression subnetwork (or cluster), while genes encoding oleosins and seed storage proteins were identified in another subnetwork with a distinct expression profile. In the triacylglycerol assembly pathway, only the genes encoding diacylglycerol acyltransferase 1 (DGAT1) and a putative cytosolic "type 3" DGAT exhibited a similar expression pattern with genes encoding oleosins. We also detected a large number of putative cis-acting regulatory elements in the promoter regions of these genes, and promoter motifs for LEC1 (LEAFY COTYLEDON 1), DOF (DNA-binding-with-One-Finger), GATA, and MYB transcription factors (TF), as well as SORLIP5 (Sequences Over Represented in Light-Induced Promoters 5), are overrepresented in the promoter regions of fatty acid biosynthetic genes. The conserved CCAAT motifs for B3 domain TFs and binding sites for bZIP (basic-leucine zipper) TFs are enriched in the promoters of genes encoding oleosins and seed storage proteins. CONCLUSIONS: Genes involved in the accumulation of seed storage reserves are expressed in distinct patterns and regulated by different TFs. The gene coexpression clusters and putative regulatory elements presented here provide a useful resource for further experimental characterization of protein interactions and regulatory networks in this process. PMID- 21635768 TI - Primary neuronal precursors in adult crayfish brain: replenishment from a non neuronal source. AB - BACKGROUND: Adult neurogenesis, the production and integration of new neurons into circuits in the brains of adult animals, is a common feature of a variety of organisms, ranging from insects and crustaceans to birds and mammals. In the mammalian brain the 1st-generation neuronal precursors, the astrocytic stem cells, reside in neurogenic niches and are reported to undergo self-renewing divisions, thereby providing a source of new neurons throughout an animal's life. In contrast, our work shows that the 1st-generation neuronal precursors in the crayfish (Procambarus clarkii) brain, which also have glial properties and lie in a neurogenic niche resembling that of vertebrates, undergo geometrically symmetrical divisions and both daughters appear to migrate away from the niche. However, in spite of this continuous efflux of cells, the number of neuronal precursors in the crayfish niche continues to expand as the animals grow and age. Based on these observations we have hypothesized that (1) the neuronal stem cells in the crayfish brain are not self-renewing, and (2) a source external to the neurogenic niche must provide cells that replenish the stem cell pool. RESULTS: In the present study, we tested the first hypothesis using sequential double nucleoside labeling to track the fate of 1st- and 2nd-generation neuronal precursors, as well as testing the size of the labeled stem cell pool following increasing incubation times in 5-bromo-2'-deoxyuridine (BrdU). Our results indicate that the 1st-generation precursor cells in the crayfish brain, which are functionally analogous to neural stem cells in vertebrates, are not a self renewing population. In addition, these studies establish the cycle time of these cells. In vitro studies examining the second hypothesis show that Cell TrackerTM Green-labeled cells extracted from the hemolymph, but not other tissues, are attracted to and incorporated into the neurogenic niche, a phenomenon that appears to involve serotonergic mechanisms. CONCLUSIONS: These results challenge our current understanding of self-renewal capacity as a defining characteristic of all adult neuronal stem cells. In addition, we suggest that in crayfish, the hematopoietic system may be a source of cells that replenish the niche stem cell pool. PMID- 21635769 TI - Untapped ethical resources for neurodegeneration research. AB - BACKGROUND: The research community has a mandate to discover effective treatments for neurodegenerative disorders. The ethics landscape surrounding this mandate is in a constant state of flux, and ongoing challenges place ever greater demands on investigators to be accountable to the public and to answer questions about the implications of their work for health care, society, and policy. METHODS: We surveyed US-based investigators involved in neurodegenerative diseases research about how they value ethics-related issues, what motivates them to give consideration to those issues, and the barriers to doing so. Using the NIH CRISP database we identified 1,034 researchers with relevant, active grants and invited them to complete an online questionnaire. We received 193 responses. We used exploratory factor analysis to transform individual survey questions into a smaller set of factors, and linear regression to understand the effect of key variables of interest on the factor scores. RESULTS: Ethics-related issues clustered into two groups: research ethics and external influences. Heads of research groups viewed issues of research ethics to be more important than the other respondents. Concern about external influences was related to overall interest in ethics. Motivators clustered into five groups: ensuring public understanding, external forces, requirements, values, and press and public. Heads of research groups were more motivated to ensure public understanding of research than the other respondents. Barriers clustered into four groups: lack of resources, administrative burden, relevance to the research, and lack of interest. Perceived lack of ethics resources was a particular barrier for investigators working in drug discovery. CONCLUSIONS: The data suggest that senior level neuroscientists working in the field of neurodegeneration (ND), and drug discovery specifically, are motivated to consider ethics issues related to their work, but the perceived lack of ethics resources thwarts their efforts. With bioethics centres at more than 50% of the institutions at which these respondents reside, the neuroscience and bioethics communities appear to be disconnected. Dedicated ethical, legal and social implications (ELSI) programs, such as those fully integrated into genetics and regenerative medicine, provide models for achieving meaningful partnerships not yet adequately realized for scholars and trainees interested in drug discovery for ND. PMID- 21635770 TI - The influence of longitudinal mentoring on medical student selection of primary care residencies. AB - BACKGROUND: The number of students selecting careers in primary care has declined by 41% in the last decade, resulting in anticipated shortages. METHODS: First year medical students interested in primary care were paired with primary care mentors. Mentors were trained, and mentors and students participated in focus groups at the end of each academic year. Quantitative and qualitative results are presented. RESULTS: Students who remained in the mentoring program matched to primary care programs at 87.5% in the first year and 78.9% in the second year, compared to overall discipline-specific match rates of 55.8% and 35.9% respectively. Students reported a better understanding of primary care and appreciated a relationship with a mentor. CONCLUSIONS: A longitudinal mentoring program can effectively support student interest in primary care if it focuses on the needs of the students and is supportive of the mentors. PMID- 21635771 TI - Clinical and technical phosphoproteomic research. AB - An encouraging approach for the diagnosis and effective therapy of immunological pathologies, which would include cancer, is the identification of proteins and phosphorylated proteins. Disease proteomics, in particular, is a potentially useful method for this purpose. A key role is played by protein phosphorylation in the regulation of normal immunology disorders and targets for several new cancer drugs and drug candidates are cancer cells and protein kinases. Protein phosphorylation is a highly dynamic process. The functioning of new drugs is of major importance as is the selection of those patients who would respond best to a specific treatment regime. In all major aspects of cellular life signalling networks are key elements which play a major role in inter- and intracellular communications. They are involved in diverse processes such as cell-cycle progression, cellular metabolism, cell-cell communication and appropriate response to the cellular environment. A whole range of networks that are involved in the regulation of cell development, differentiation, proliferation, apoptosis, and immunologic responses is contained in the latter. It is so necessary to understand and monitor kinase signalling pathways in order to understand many immunology pathologies. Enrichment of phosphorylated proteins or peptides from tissue or bodily fluid samples is required. The application of technologies such as immunoproteomic techniques, phosphoenrichments and mass spectrometry (MS) is crucial for the identification and quantification of protein phosphorylation sites in order to advance in clinical research. Pharmacodynamic readouts of disease states and cellular drug responses in tumour samples will be provided as the field develops. We aim to detail the current and most useful techniques with research examples to isolate and carry out clinical phosphoproteomic studies which may be helpful for immunology and cancer research. Different phosphopeptide enrichment and quantitative techniques need to be combined to achieve good phosphopeptide recovery and good up- and-down phospho-regulation protein studies. PMID- 21635772 TI - Metabolic and endocrine profiles and reproductive parameters in dairy cows under grazing conditions: effect of polymorphisms in somatotropic axis genes. AB - BACKGROUND: The present study hypothesized that GH-AluI and IGF-I-SnabI polymorphisms do change the metabolic/endocrine profiles in Holstein cows during the transition period, which in turn are associated with productive and reproductive parameters. METHODS: Holstein cows (Farm 1, primiparous cows, n = 110, and Farm 2, multiparous cows, n = 76) under grazing conditions were selected and GH and IGF-I genotypes were determined. Blood samples for metabolic/endocrine determinations were taken during the transition period and early lactation in both farms. Data was analyzed by farm using a repeated measures analyses including GH and IGF-I genotypes, days and interactions as fixed effects, sire and cow as random effects and calving date as covariate. RESULTS AND DISCUSSION: Frequencies of GH and IGF-I alleles were L:0.84, V:0.16 and A:0.60, B:0.40, respectively. The GH genotype was not associated with productive or reproductive variables, but interaction with days affected FCM yield in multiparous (farm 2) cows (LL yielded more than LV cows) in early lactation. The GH genotype affected NEFA and IGF-I concentrations in farm 1 (LV had higher NEFA and lower IGF-I than LL cows) suggesting a better energy status of LL cows.There was no effect of IGF I genotype on productive variables, but a trend was found for FCM in farm 2 (AB cows yielded more than AA cows). IGF-I genotype affected calving first service interval in farm 1, and the interaction with days tended to affect FCM yield (AB cows had a shorter interval and yielded more FCM than BB cows). IGF-I genotype affected BHB, NEFA, and insulin concentrations in farm 1: primiparous BB cows had lower NEFA and BHB and higher insulin concentrations. In farm 2, there was no effect of IGF-I genotype, but there was an interaction with days on IGF-I concentration, suggesting a greater uncoupling somatropic axis in AB and BB than AA cows, being in accordance with greater FCM yield in AB cows. CONCLUSION: The GH and IGF-I genotypes had no substantial effect on productive parameters, although IGF-I genotype affected calving-first service interval in primiparous cows. Besides, these genotypes may modify the endocrine/metabolic profiles of the transition dairy cow under grazing conditions. PMID- 21635773 TI - Methylenetetrahydrofolate reductase gene variant (MTHFR C677T) and migraine: a case control study and meta-analysis. AB - BACKGROUND: Migraine is a common disorder that often coexists with depression. While a functional polymorphism in methyleneterahydrofolate reductase gene (MTHFR C677T) has been implicated in depression; the evidence to support an association of MTHFR with migraine has been inconclusive. We aim to investigate the effect of this variant on propensity for migraine and to perform a systematic review and meta-analysis of studies of MTHFR and migraine to date. METHODS: Individuals with migraine (n = 447) were selected from the Depression Case Control (DeCC) study to investigate the association between migraine and MTHFR C677T single nucleotide polymorphism (SNP) rs1801133 using an additive model compared to non-migraineurs adjusting for depression status. A meta-analysis was performed and included 15 studies of MTHFR and migraine. RESULTS: MTHFR C677T polymorphism was associated with migraine with aura (MA) (OR 1.31, 95% CI 1.01-1.70, p = 0.039) that remained significant after adjusting for age, sex and depression status. A meta-analysis of 15 case-control studies showed that T allele homozygosity is significantly associated with MA (OR = 1.42; 95% CI, 1.10-1.82) and total migraine (OR = 1.37; 95% CI, 1.07-1.76), but not migraine without aura (OR = 1.16; 95% CI, 0.36-3.76). In studies of non-Caucasian population, the TT genotype was associated with total migraine (OR= 3.46; 95% CI, 1.22-9.82), whereas in studies of Caucasians this variant was associated with MA only (OR = 1.28; 95% CI, 1.002-1.63). CONCLUSIONS: MTHFR C677T is associated with MA in individuals selected for depression study. A meta-analysis of 15 studies supports this association and demonstrated effects across ethnic groups. PMID- 21635774 TI - Impact of changing the measles vaccine vial size on Niger's vaccine supply chain: a computational model. AB - BACKGROUND: Many countries, such as Niger, are considering changing their vaccine vial size presentation and may want to evaluate the subsequent impact on their supply chains, the series of steps required to get vaccines from their manufacturers to patients. The measles vaccine is particularly important in Niger, a country prone to measles outbreaks. METHODS: We developed a detailed discrete event simulation model of the vaccine supply chain representing every vaccine, storage location, refrigerator, freezer, and transport device (e.g., cold trucks, 4 * 4 trucks, and vaccine carriers) in the Niger Expanded Programme on Immunization (EPI). Experiments simulated the impact of replacing the 10-dose measles vial size with 5-dose, 2-dose and 1-dose vial sizes. RESULTS: Switching from the 10-dose to the 5-dose, 2-dose and 1-dose vial sizes decreased the average availability of EPI vaccines for arriving patients from 83% to 82%, 81% and 78%, respectively for a 100% target population size. The switches also changed transport vehicle's utilization from a mean of 58% (range: 4-164%) to means of 59% (range: 4-164%), 62% (range: 4-175%), and 67% (range: 5-192%), respectively, between the regional and district stores, and from a mean of 160% (range: 83-300%) to means of 161% (range: 82-322%), 175% (range: 78-344%), and 198% (range: 88-402%), respectively, between the district to integrated health centres (IHC). The switch also changed district level storage utilization from a mean of 65% to means of 64%, 66% and 68% (range for all scenarios: 3-100%). Finally, accounting for vaccine administration, wastage, and disposal, replacing the 10-dose vial with the 5 or 1-dose vials would increase the cost per immunized patient from $0.47US to $0.71US and $1.26US, respectively. CONCLUSIONS: The switch from the 10-dose measles vaccines to smaller vial sizes could overwhelm the capacities of many storage facilities and transport vehicles as well as increase the cost per vaccinated child. PMID- 21635775 TI - Focal laser ablation of prostate cancer: numerical simulation of temperature and damage distribution. AB - BACKGROUND: The use of minimally invasive ablative techniques in the management of patients with low grade and localized prostate tumours could represent a treatment option between active surveillance and radical therapy. Focal laser ablation (FLA) could be one of these treatment modalities. Dosimetry planning and conformation of the treated area to the tumor remain major issues, especially when, several fibers are required. An effective method to perform pre-treatment planning of this therapy is computer simulation. In this study we present an in vivo validation of a mathematical model. METHODS: The simulation model is based on finite elements method (FEM) to solve the bio-heat and the thermal damage equations. Laser irradiation was performed with a 980 nm laser diode system (5 W, 75 s). Light was transmitted using a cylindrical diffusing fiber inserted inside a preclinical animal prostate cancer model induced in Copenhagen rats. Non enhanced T2-weighted and dynamic gadolinium-enhanced T1-weighted MR imaging examinations were performed at baseline and 48 hours after the procedure. The model was validated by comparing the simulated necrosis volume to the results obtained in vivo on (MRI) and by histological analysis. 3 iso-damage temperatures were considered 43 degrees C, 45 degrees C and 50 degrees C. RESULTS: The mean volume of the tissue necrosis, estimated from the histological analyses was 0.974 +/- 0.059 cc and 0.98 +/- 0.052 cc on the 48 h MR images. For the simulation model, volumes were: 1.38 cc when T = 43 degrees C, 1.1 cc for T = 45 degrees C and 0.99 cc when T = 50 C degrees . CONCLUSIONS: In this study, a clear correlation was established between simulation and in vivo experiments of FLA for prostate cancer.Simulation is a promising planning technique for this therapy. It needs further more evaluation to allow to FLA to become a widely applied surgical method. PMID- 21635776 TI - Low early ototoxicity rates for pediatric medulloblastoma patients treated with proton radiotherapy. AB - BACKGROUND: Hearing loss is common following chemoradiotherapy for children with medulloblastoma. Compared to photons, proton radiotherapy reduces radiation dose to the cochlea for these patients. Here we examine whether this dosimetric advantage leads to a clinical benefit in audiometric outcomes. METHODS: From 2006 2009, 23 children treated with proton radiotherapy for medulloblastoma were enrolled on a prospective observational study, through which they underwent pre- and 1 year post-radiotherapy pure-tone audiometric testing. Ears with moderate to severe hearing loss prior to therapy were censored, leaving 35 ears in 19 patients available for analysis. RESULTS: The predicted mean cochlear radiation dose was 30 60Co-Gy Equivalents (range 19-43), and the mean cumulative cisplatin dose was 303 mg/m2 (range 298-330). Hearing sensitivity significantly declined following radiotherapy across all frequencies analyzed (P < 0.05). There was partial sparing of mean post-radiation hearing thresholds at low-to-midrange frequencies and, consequently, the rate of high-grade (grade 3 or 4) ototoxicity at 1 year was favorable (5%). Ototoxicity did not correlate with predicted dose to the auditory apparatus for proton-treated patients, potentially reflecting a lower-limit threshold for radiation effect on the cochlea. CONCLUSIONS: Rates of high-grade early post-radiation ototoxicity following proton radiotherapy for pediatric medulloblastoma are low. Preservation of hearing in the audible speech range, as observed here, may improve both quality of life and cognitive functioning for these patients. PMID- 21635777 TI - Sclerosing Mesenteritis: Diverse clinical presentations and dissimilar treatment options. A case series and review of the literature. AB - Sclerosing mesenteritis (SM) is a rare pathological condition affecting the mesentery. It is a benign, non-specific inflammation of the adipose tissue of the mesentery of the small intestine and colon. It is characterized by a variable amount of chronic fibrosis. Its etiology is unknown, the pathogenesis is obscure, while the pathological characteristics of the disease are unspecific. The initial clinical presentation varies from typically asymptomatic to that of an acute abdomen. The diagnosis is suggested by computed tomography but is usually confirmed by surgical biopsies. Treatment is largely empirical; it is decided upon on the basis of the clinical condition of the patient, and usually a few specific drugs are used. Surgical resection is sometimes attempted for definitive therapy, although the surgical approach is often limited. We will present five cases of SM as well as a review of the available literature in order to state and compare a variety of clinical presentations, diverse possible etiologies and dissimilar treatment options. PMID- 21635778 TI - Genetic basis of wing morphogenesis in Drosophila: sexual dimorphism and non allometric effects of shape variation. AB - BACKGROUND: The Drosophila wing represents a particularly appropriate model to investigate the developmental control of phenotypic variation. Previous studies which aimed to identify candidate genes for wing morphology demonstrated that the genetic basis of wing shape variation in D. melanogaster is composed of numerous genetic factors causing small, additive effects. In this study, we analyzed wing shape in males and females from 191 lines of D. melanogaster, homozygous for a single P-element insertion, using geometric morphometrics techniques. The analysis allowed us to identify known and novel candidate genes that may contribute to the expression of wing shape in each sex separately and to compare them to candidate genes affecting wing size which have been identified previously using the same lines. RESULTS: Our results indicate that more than 63% of induced mutations affected wing shape in one or both sexes, although only 33% showed significant differences in both males and females. The joint analysis of wing size and shape revealed that only 19% of the P-element insertions caused coincident effects on both components of wing form in one or both sexes. Further morphometrical analyses revealed that the intersection between veins showed the smallest displacements in the proximal region of the wing. Finally, we observed that mutations causing general deformations were more common than expected in both sexes whereas the opposite occurred with those generating local changes. For most of the 94 candidate genes identified, this seems to be the first record relating them with wing shape variation. CONCLUSIONS: Our results support the idea that the genetic architecture of wing shape is complex with many different genes contributing to the trait in a sexually dimorphic manner. This polygenic basis, which is relatively independent from that of wing size, is composed of genes generally involved in development and/or metabolic functions, especially related to the regulation of different cellular processes such as motility, adhesion, communication and signal transduction. This study suggests that understanding the genetic basis of wing shape requires merging the regulation of vein patterning by signalling pathways with processes that occur during wing development at the cellular level. PMID- 21635779 TI - mRNA binding protein staufen 1-dependent regulation of pyramidal cell spine morphology via NMDA receptor-mediated synaptic plasticity. AB - Staufens (Stau) are RNA-binding proteins involved in mRNA transport, localization, decay and translational control. The Staufen 1 (Stau1) isoform was recently identified as necessary for the protein synthesis-dependent late phase long-term potentiation (late-LTP) and for the maintenance of mature dendritic spines and synaptic activity in hippocampal CA1 pyramidal cells, strongly suggesting a role of mRNA regulation by Stau1 in these processes. However, the causal relationship between these impairments in synaptic function (spine shape and basal synaptic activity) and plasticity (late-LTP) remains unclear. Here, we determine that the effects of Stau1 knockdown on spine shape and size are mimicked by blocking NMDA receptors (or elevating extracellular Mg2+) and that Stau1 knockdown in the presence of NMDA receptor blockade (or high Mg2+) has no further effect on spine shape and size. Moreover, the effect of Stau1 knockdown on late-LTP cannot be explained by these effects, since when tested in normal medium, slice cultures that had been treated with high Mg2+ (to impair NMDA receptor function) in combination with a control siRNA still exhibited late-LTP, while siRNA to Stau1 was still effective in blocking late-LTP. Our results indicate that Stau1 involvement in spine morphogenesis is dependent on ongoing NMDA receptor-mediated plasticity, but its effects on late-LTP are independent of these changes. These findings clarify the role of Stau1-dependent mRNA regulation in physiological and morphological changes underlying long-term synaptic plasticity in pyramidal cells. PMID- 21635780 TI - Site-specific mobilization of vinyl chloride respiration islands by a mechanism common in Dehalococcoides. AB - BACKGROUND: Vinyl chloride is a widespread groundwater pollutant and Group 1 carcinogen. A previous comparative genomic analysis revealed that the vinyl chloride reductase operon, vcrABC, of Dehalococcoides sp. strain VS is embedded in a horizontally-acquired genomic island that integrated at the single-copy tmRNA gene, ssrA. RESULTS: We targeted conserved positions in available genomic islands to amplify and sequence four additional vcrABC -containing genomic islands from previously-unsequenced vinyl chloride respiring Dehalococcoides enrichments. We identified a total of 31 ssrA-specific genomic islands from Dehalococcoides genomic data, accounting for 47 reductive dehalogenase homologous genes and many other non-core genes. Sixteen of these genomic islands contain a syntenic module of integration-associated genes located adjacent to the predicted site of integration, and among these islands, eight contain vcrABC as genetic 'cargo'. These eight vcrABC -containing genomic islands are syntenic across their ~12 kbp length, but have two phylogenetically discordant segments that unambiguously differentiate the integration module from the vcrABC cargo. Using available Dehalococcoides phylogenomic data we estimate that these ssrA-specific genomic islands are at least as old as the Dehalococcoides group itself, which in turn is much older than human civilization. CONCLUSIONS: The vcrABC -containing genomic islands are a recently-acquired subset of a diverse collection of ssrA specific mobile elements that are a major contributor to strain-level diversity in Dehalococcoides, and may have been throughout its evolution. The high similarity between vcrABC sequences is quantitatively consistent with recent horizontal acquisition driven by ~100 years of industrial pollution with chlorinated ethenes. PMID- 21635781 TI - Diverse and tissue-enriched small RNAs in the plant pathogenic fungus, Magnaporthe oryzae. AB - BACKGROUND: Emerging knowledge of the impact of small RNAs as important cellular regulators has prompted an explosion of small transcriptome sequencing projects. Although significant progress has been made towards small RNA discovery and biogenesis in higher eukaryotes and other model organisms, knowledge in simple eukaryotes such as filamentous fungi remains limited. RESULTS: Here, we used 454 pyrosequencing to present a detailed analysis of the small RNA transcriptome (~ 15 - 40 nucleotides in length) from mycelia and appressoria tissues of the rice blast fungal pathogen, Magnaporthe oryzae. Small RNAs mapped to numerous nuclear and mitochondrial genomic features including repetitive elements, tRNA loci, rRNAs, protein coding genes, snRNAs and intergenic regions. For most elements, small RNAs mapped primarily to the sense strand with the exception of repetitive elements to which small RNAs mapped in the sense and antisense orientation in near equal proportions. Inspection of the small RNAs revealed a preference for U and suppression of C at position 1, particularly for antisense mapping small RNAs. In the mycelia library, small RNAs of the size 18 - 23 nt were enriched for intergenic regions and repetitive elements. Small RNAs mapping to LTR retrotransposons were classified as LTR retrotransposon-siRNAs (LTR-siRNAs). Conversely, the appressoria library had a greater proportion of 28 - 35 nt small RNAs mapping to tRNA loci, and were classified as tRNA-derived RNA fragments (tRFs). LTR-siRNAs and tRFs were independently validated by 3' RACE PCR and northern blots, respectively. CONCLUSIONS: Our findings suggest M. oryzae small RNAs differentially accumulate in vegetative and specialized-infection tissues and may play an active role in genome integrity and regulating growth and development. PMID- 21635782 TI - Sputum and nasal lavage lung-specific biomarkers before and after smoking cessation. AB - BACKGROUND: Little is known about the effect of smoking cessation on airway inflammation. Secretory Leukocyte Protease Inhibitor (SLPI), Clara Cell protein 16 (CC16), elafin and human defensin beta-2 (HBD-2) protect human airways against inflammation and oxidative stress. In this longitudinal study we aimed to investigate changes in sputum and nasal lavage SLPI, CC16, elafin and HBD-2 levels in healthy smokers after 6 and 12 months of smoking cessation. METHODS: Induced sputum and nasal lavage was obtained from healthy current smokers (n = 76) before smoking cessation, after 6 months of smoking cessation (n = 29), after 1 year of smoking cessation (n = 22) and from 10 healthy never smokers. SLPI, CC16, elafin and HBD-2 levels were measured in sputum and nasal lavage supernatants by commercially available ELISA kits. RESULTS: Sputum SLPI and CC-16 levels were increased in healthy smokers before smoking cessation versus never smokers (p = 0.005 and p = 0.08 respectively). SLPI and CC16 levels did not differ before and 6 months after smoking cessation (p = 0.118 and p = 0.543 respectively), neither before and 1 year after smoking cessation (p = 0.363 and p = 0.470 respectively). Nasal lavage SLPI was decreased 12 months after smoking cessation (p = 0.033). Nasal lavage elafin levels were increased in healthy smokers before smoking cessation versus never-smokers (p = 0.007), but there were no changes 6 months and 1 year after smoking cessation. CONCLUSIONS: Only nasal lavage SLPI decrease after 1 year after smoking cessation. We may speculate that there is an ongoing inflammatory process stimulating the production of counter regulating proteins in the airways of healthy ex-smokers. PMID- 21635783 TI - FLIP (Flice-like inhibitory protein) suppresses cytoplasmic double-stranded-RNA induced apoptosis and NF-kappaB and IRF3-mediated signaling. AB - BACKGROUND: Cytoplasmic viral double-stranded RNA (dsRNA) is detected by a class of ubiquitous cytoplasmic RNA helicases, retinoic acid inducible gene-I (RIG-I) and melanoma differentiation antigen-5 (MDA5), which initiate a signaling cascade via their common adaptor called interferon-beta (IFN-beta) promoter stimulator-1 (IPS-1). This leads to the production of proinflammatory and antiviral cytokines, the type I Interferons, via mainly nuclear factor kappa B (NF-kappaB) and interferon response factor-3 (IRF3) transcription factors. Fas-associated death domain (FADD) protein, receptor-interacting protein (RIP1), caspase-8 and tumor necrosis factor receptor (TNFR)-associated death domain (TRADD) protein, all traditionally associated with death receptor signaling, are also involved in RIG I/MDA5 signaling pathway. We previously showed that FLIP (Flice-like inhibitory protein), also designated as cflar (CASP8 and FADD-like apoptosis regulator), negatively regulates lipopolysaccharide (LPS)-induced toll-like receptor 4 (TLR4) signaling in endothelial cells and mouse embryonic fibroblasts (MEFs) and protected against TLR4-mediated apoptosis. RESULTS: In this study, we investigated the role of FLIP in cellular response to cytoplasmic polyinosinic:polycytidylic acid, poly(I:C), a synthetic analog of dsRNA. Consistent with the previously described role of FADD in RIG-I/MDA5-mediated apoptosis, we found that FLIP-/- MEFs were more susceptible to killing by cytoplasmic poly(I:C). However, FLIP-/- MEFs also exhibited markedly increased expression of NF-kappaB-and IRF3- dependent genes in response to cytoplasmic poly(I:C). Importantly, reconstitution of FLIP in FLIP-/-MEFs reversed the hyper activation of IRF3- and NF-kappaB-mediated gene expression. Further, we found that caspase-8 catalytic activity was not required for cytoplasmic poly(I:C) mediated NF-kappaB and IRF3 signaling. CONCLUSIONS: These results provide evidence for a crucial dual role for FLIP in antiviral responses to cytoplasmic dsRNA: it protects from cytoplasmic dsRNA-mediated cell death while down regulating IRF3-and NF-kappaB-mediated gene expression. Since the pathogenesis of several viral infections involves a heightened and dysregulated cytokine response, a possible therapy could involve modulating FLIP levels. PMID- 21635784 TI - Health impact assessment of waste management facilities in three European countries. AB - BACKGROUND: Policies on waste disposal in Europe are heterogeneous and rapidly changing, with potential health implications that are largely unknown. We conducted a health impact assessment of landfilling and incineration in three European countries: Italy, Slovakia and England. METHODS: A total of 49 (Italy), 2 (Slovakia), and 11 (England) incinerators were operating in 2001 while for landfills the figures were 619, 121 and 232, respectively. The study population consisted of residents living within 3 km of an incinerator and 2 km of a landfill. Excess risk estimates from epidemiological studies were used, combined with air pollution dispersion modelling for particulate matter (PM10) and nitrogen dioxide (NO2). For incinerators, we estimated attributable cancer incidence and years of life lost (YoLL), while for landfills we estimated attributable cases of congenital anomalies and low birth weight infants. RESULTS: About 1,000,000, 16,000, and 1,200,000 subjects lived close to incinerators in Italy, Slovakia and England, respectively. The additional contribution to NO2 levels within a 3 km radius was 0.23, 0.15, and 0.14 MUg/m3, respectively. Lower values were found for PM10. Assuming that the incinerators continue to operate until 2020, we are moderately confident that the annual number of cancer cases due to exposure in 2001-2020 will reach 11, 0, and 7 in 2020 and then decline to 0 in the three countries in 2050. We are moderately confident that by 2050, the attributable impact on the 2001 cohort of residents will be 3,621 (Italy), 37 (Slovakia) and 3,966 (England) YoLL. The total exposed population to landfills was 1,350,000, 329,000, and 1,425,000 subjects, respectively. We are moderately confident that the annual additional cases of congenital anomalies up to 2030 will be approximately 2, 2, and 3 whereas there will be 42, 13, and 59 additional low-birth weight newborns, respectively. CONCLUSIONS: The current health impacts of landfilling and incineration can be characterized as moderate when compared to other sources of environmental pollution, e.g. traffic or industrial emissions, that have an impact on public health. There are several uncertainties and critical assumptions in the assessment model, but it provides insight into the relative health impact attributable to waste management. PMID- 21635785 TI - Machado-Joseph Disease: from first descriptions to new perspectives. AB - Machado-Joseph Disease (MJD), also known as spinocerebellar ataxia type 3 (SCA3), represents the most common form of SCA worldwide. MJD is an autosomal dominant neurodegenerative disorder of late onset, involving predominantly the cerebellar, pyramidal, extrapyramidal, motor neuron and oculomotor systems; although sharing features with other SCAs, the identification of minor, but more specific signs, facilitates its differential diagnosis. MJD presents strong phenotypic heterogeneity, which has justified the classification of patients into three main clinical types. Main pathological lesions are observed in the spinocerebellar system, as well as in the cerebellar dentate nucleus. MJD's causative mutation consists in an expansion of an unstable CAG tract in exon 10 of the ATXN3 gene, located at 14q32.1. Haplotype-based studies have suggested that two main founder mutations may explain the present global distribution of the disease; the ancestral haplotype is of Asian origin, and has an estimated age of around 5,800 years, while the second mutational event has occurred about 1,400 years ago. The ATXN3 gene encodes for ataxin-3, which is ubiquitously expressed in neuronal and non-neuronal tissues, and, among other functions, is thought to participate in cellular protein quality control pathways. Mutated ATXN3 alleles consensually present about 61 to 87 CAG repeats, resulting in an expanded polyglutamine tract in ataxin-3. This altered protein gains a neurotoxic function, through yet unclear mechanisms. Clinical variability of MJD is only partially explained by the size of the CAG tract, which leaves a residual variance that should be explained by still unknown additional factors. Several genetic tests are available for MJD, and Genetic Counseling Programs have been created to better assist the affected families, namely on what concerns the possibility of pre symptomatic testing. The main goal of this review was to bring together updated knowledge on MJD, covering several aspects from its initial descriptions and clinical presentation, through the discovery of the causative mutation, its origin and dispersion, as well as molecular genetics aspects considered essential for a better understanding of its neuropathology. Issues related with molecular testing and Genetic Counseling, as well as recent progresses and perspectives on genetic therapy, are also addressed. PMID- 21635786 TI - StralSV: assessment of sequence variability within similar 3D structures and application to polio RNA-dependent RNA polymerase. AB - BACKGROUND: Most of the currently used methods for protein function prediction rely on sequence-based comparisons between a query protein and those for which a functional annotation is provided. A serious limitation of sequence similarity based approaches for identifying residue conservation among proteins is the low confidence in assigning residue-residue correspondences among proteins when the level of sequence identity between the compared proteins is poor. Multiple sequence alignment methods are more satisfactory--still, they cannot provide reliable results at low levels of sequence identity. Our goal in the current work was to develop an algorithm that could help overcome these difficulties by facilitating the identification of structurally (and possibly functionally) relevant residue-residue correspondences between compared protein structures. RESULTS: Here we present StralSV (structure-alignment sequence variability), a new algorithm for detecting closely related structure fragments and quantifying residue frequency from tight local structure alignments. We apply StralSV in a study of the RNA-dependent RNA polymerase of poliovirus, and we demonstrate that the algorithm can be used to determine regions of the protein that are relatively unique, or that share structural similarity with proteins that would be considered distantly related. By quantifying residue frequencies among many residue-residue pairs extracted from local structural alignments, one can infer potential structural or functional importance of specific residues that are determined to be highly conserved or that deviate from a consensus. We further demonstrate that considerable detailed structural and phylogenetic information can be derived from StralSV analyses. CONCLUSIONS: StralSV is a new structure based algorithm for identifying and aligning structure fragments that have similarity to a reference protein. StralSV analysis can be used to quantify residue-residue correspondences and identify residues that may be of particular structural or functional importance, as well as unusual or unexpected residues at a given sequence position. StralSV is provided as a web service at http://proteinmodel.org/AS2TS/STRALSV/. PMID- 21635787 TI - Patterns of multimorbidity in working Australians. AB - BACKGROUND: Multimorbidity is becoming more prevalent. Previously-used methods of assessing multimorbidity relied on counting the number of health conditions, often in relation to an index condition (comorbidity), or grouping conditions based on body or organ systems. Recent refinements in statistical approaches have resulted in improved methods to capture patterns of multimorbidity, allowing for the identification of nonrandomly occurring clusters of multimorbid health conditions. This paper aims to identify nonrandom clusters of multimorbidity. METHODS: The Australian Work Outcomes Research Cost-benefit (WORC) study cross sectional screening dataset (approximately 78,000 working Australians) was used to explore patterns of multimorbidity. Exploratory factor analysis was used to identify nonrandomly occurring clusters of multimorbid health conditions. RESULTS: Six clinically-meaningful groups of multimorbid health conditions were identified. These were: factor 1: arthritis, osteoporosis, other chronic pain, bladder problems, and irritable bowel; factor 2: asthma, chronic obstructive pulmonary disease, and allergies; factor 3: back/neck pain, migraine, other chronic pain, and arthritis; factor 4: high blood pressure, high cholesterol, obesity, diabetes, and fatigue; factor 5: cardiovascular disease, diabetes, fatigue, high blood pressure, high cholesterol, and arthritis; and factor 6: irritable bowel, ulcer, heartburn, and other chronic pain. These clusters do not fall neatly into organ or body systems, and some conditions appear in more than one cluster. CONCLUSIONS: Considerably more research is needed with large population-based datasets and a comprehensive set of reliable health diagnoses to better understand the complex nature and composition of multimorbid health conditions. PMID- 21635788 TI - Research in advance for FMD novel vaccines. AB - Foot-and-Mouth Disease (FMD), as a major global animal disease, affects millions of animals worldwide and remains the main sanitary barrier to the international and national trade of animals and animal products. Inactivated vaccination is the most effective measure for prevention of FMD at present, but fail to induce long term protection and content new requires for production of FMD vaccines. As a number of Researchers hope to obtain satisfactory novel vaccines by new bio technology, novel vaccines have been studied for more than thirty years. Here reviews the latest research progress of new vaccines, summarizes some importance and raises several suggestions for the future of FMD vaccine. PMID- 21635789 TI - Self-organization of developing embryo using scale-invariant approach. AB - BACKGROUND: Self-organization is a fundamental feature of living organisms at all hierarchical levels from molecule to organ. It has also been documented in developing embryos. METHODS: In this study, a scale-invariant power law (SIPL) method has been used to study self-organization in developing embryos. The SIPL coefficient was calculated using a centro-axial skew symmetrical matrix (CSSM) generated by entering the components of the Cartesian coordinates; for each component, one CSSM was generated. A basic square matrix (BSM) was constructed and the determinant was calculated in order to estimate the SIPL coefficient. This was applied to developing C. elegans during early stages of embryogenesis. The power law property of the method was evaluated using the straight line and Koch curve and the results were consistent with fractal dimensions (fd). Diffusion-limited aggregation (DLA) was used to validate the SIPL method. RESULTS AND CONCLUSION: The fractal dimensions of both the straight line and Koch curve showed consistency with the SIPL coefficients, which indicated the power law behavior of the SIPL method. The results showed that the ABp sublineage had a higher SIPL coefficient than EMS, indicating that ABp is more organized than EMS. The fd determined using DLA was higher in ABp than in EMS and its value was consistent with type 1 cluster formation, while that in EMS was consistent with type 2. PMID- 21635790 TI - Profile of mood states and stress-related biochemical indices in long-term yoga practitioners. AB - BACKGROUND: Previous studies have shown the short-term or intermediate-term practice of yoga to be useful for ameliorating several mental disorders and psychosomatic disorders. However, little is known about the long-term influences of yoga on the mental state or stress-related biochemical indices. If yoga training has a stress-reduction effect and also improves an individual's mental states for a long time, long-term yoga practitioners may have a better mental state and lower stress-related biochemical indices in comparison to non experienced participants. This study simultaneously examined the differences in mental states and urinary stress-related biochemical indices between long-term yoga practitioners and non-experienced participants. METHODS: The participants were 38 healthy females with more than 2 years of experience with yoga (long-term yoga group) and 37 age-matched healthy females who had not participated in yoga (control group). Their mental states were assessed using the Profile of Mood States (POMS) questionnaire. The level of cortisol, 8-hydroxydeoxyguanosine (8 OHdG) and biopyrrin in urine were used as stress-related biochemical indices. RESULTS: The average self-rated mental disturbance, tension-anxiety, anger hostility, and fatigue scores of the long-term yoga group were lower than those of the control group. There was a trend toward a higher vigor score in the long term yoga group than that in the control group. There were no significant differences in the scores for depression and confusion in the POMS between the two groups. The urine 8-OHdG concentration showed a trend toward to being lower in the long-term yoga group in comparison to the control group. There were no significant differences in the levels of urine biopyrrin or cortisol. CONCLUSIONS: The present findings suggest that long-term yoga training can reduce the scores related to mental health indicators such as self-rated anxiety, anger, and fatigue. PMID- 21635791 TI - Community interventions to reduce child mortality in Dhanusha, Nepal: study protocol for a cluster randomized controlled trial. AB - BACKGROUND: Neonatal mortality remains high in rural Nepal. Previous work suggests that local women's groups can effect significant improvement through community mobilisation. The possibility of identification and management of newborn infections by community-based workers has also arisen. METHODS/DESIGN: The objective of this trial is to evaluate the effects on newborn health of two community-based interventions involving Female Community Health Volunteers. MIRA Dhanusha community groups: a participatory intervention with women's groups. MIRA Dhanusha sepsis management: training of community volunteers in the recognition and management of neonatal sepsis.The study design is a cluster randomized controlled trial involving 60 village development committee clusters allocated 1:1 to two interventions in a factorial design. MIRA Dhanusha community groups: Female Community Health Volunteers (FCHVs) are supported in convening monthly women's groups. Nine groups per cluster (270 in total) work through two action research cycles in which they (i) identify local issues around maternity, newborn health and nutrition, (ii) prioritize key problems, (iii) develop strategies to address them, (iv) implement the strategies, and (v) evaluate their success. Cycle 1 focuses on maternal and newborn health and cycle 2 on nutrition in pregnancy and infancy and associated postpartum care practices. MIRA Dhanusha sepsis management: FCHVs are trained to care for vulnerable newborn infants. They (i) identify local births, (ii) identify low birth weight infants, (iii) identify possible newborn infection, (iv) manage the process of treatment with oral antibiotics and referral to a health facility to receive parenteral gentamicin, and (v) follow up infants and support families. PRIMARY OUTCOME: neonatal mortality rates. SECONDARY OUTCOMES: MIRA Dhanusha community group: stillbirth, infant and under-two mortality rates, care practices and health care seeking behaviour, maternal diet, breastfeeding and complementary feeding practices, maternal and under-2 anthropometric status. MIRA Dhanusha sepsis management: identification and treatment of neonatal sepsis by community health volunteers, infection-specific neonatal mortality. PMID- 21635792 TI - Oncogenic potential of Human Papillomavirus (HPV) and its relation with cervical cancer. AB - Human Papillomavirus (HPV) is the most common cause of cervical cancer. Cervical cancer being the second most common cancer after lung cancer, affecting women of different age groups; has a prevalence of about 20% in young sexually active women. Among different types of HPV, HPV16 the major strain causing this cancer and is sexually transmitted had been unnoticed for decades. Keeping in mind the multiple risk factors related with cervical cancer such as early age sexual activities, teenage pregnancies, smoking, use of oral contraceptives, having multiple sex partners, hormone replacement therapies and various other unknown factors lead to the onset of the disease. Awareness for various diagnostic procedures such as Pap smears screening prove to be an effective way in eradicating the oncogenic potential of HPV. PMID- 21635793 TI - Ultrasound in rheumatoid arthritis: volar versus dorsal synovitis evaluation and scoring. AB - BACKGROUND: Assessment of synovitis in rheumatoid arthritis (RA) is a major issue for a proper treatment administration; it has been proven that ultrasound (US) examination could be of valuable help and it is currently being investigated as a possible outcome measure for the disease. It is, though, of greatest importance to accurately establish the place of US scores among the already validated outcome measures, according to Outcome Measures for Rheumatoid Arthritis in Clinical Trials (OMERACT) filter. The present study is designed to compare the results of gray-scale ultrasound (GSUS) and Power Doppler ultrasound (PDUS) additive scores, separately calculated for volar and dorsal aspects of the hand, with physical examination, patient's evaluation of disease pain and global activity on Visual Analogic Scale (VAS) and traditional scores for disease activity assessment (DAS28, CDAI, SDAI, HAQ). The final aim is to prove the advantages of volar US evaluation in RA patients. METHODS: 42 RA patients have been clinically evaluated for pain and swelling of their hand joints, completed VAS and HAQ questionnaires and underwent both volar and dorsal sonography of the hands during the same day. The US examiner was blinded to clinical assessments and lab results. For each patient 20 joints were assessed by sonography (radiocarpal, intercarpal, metacarpophalangeal (MCP) 2-5, proximal interphalangeal (PIP) 2-5). Carpal joints were only evaluated from dorsal view, while MCPs and PIPs were evaluated both from dorsal and volar aspect resulting a total of 36 distinct evaluations for each patient. GSUS synovial hypertrophy was assessed both by quantitative measurement and semiquantitative scale (0-3 grades); Doppler signal (PDUS) was recorded on a semiquantitative scale (0-3 grades). The semiquantitative grades for both GSUS and PDUS evaluation of each joint were added and the sum was defined as the Echographic Score (ES) of each patient. Separately, we added the semiquantitative grades for volar and dorsal side, resulting in Volar ES (VES) and Dorsal ES (DES) of each patient. RESULTS: We found ESs correlated with other activity scores: DAS28, CDAI, SDAI, HAQ. Correlations with clinical indices as CDAI and SDAI were stronger for VES than for DES. US discovered more synovitis than clinical examination. CONCLUSION: VES is a suitable reflection of RA activity and volar US examination should accompany the dorsal one both in clinical practice and in clinical trials. PMID- 21635794 TI - The impact of Ty3-gypsy group LTR retrotransposons Fatima on B-genome specificity of polyploid wheats. AB - BACKGROUND: Transposable elements (TEs) are a rapidly evolving fraction of the eukaryotic genomes and the main contributors to genome plasticity and divergence. Recently, occupation of the A- and D-genomes of allopolyploid wheat by specific TE families was demonstrated. Here, we investigated the impact of the well represented family of gypsy LTR-retrotransposons, Fatima, on B-genome divergence of allopolyploid wheat using the fluorescent in situ hybridisation (FISH) method and phylogenetic analysis. RESULTS: FISH analysis of a BAC clone (BAC_2383A24) initially screened with Spelt1 repeats demonstrated its predominant localisation to chromosomes of the B-genome and its putative diploid progenitor Aegilops speltoides in hexaploid (genomic formula, BBAADD) and tetraploid (genomic formula, BBAA) wheats as well as their diploid progenitors. Analysis of the complete BAC_2383A24 nucleotide sequence (113,605 bp) demonstrated that it contains 55.6% TEs, 0.9% subtelomeric tandem repeats (Spelt1), and five genes. LTR retrotransposons are predominant, representing 50.7% of the total nucleotide sequence. Three elements of the gypsy LTR retrotransposon family Fatima make up 47.2% of all the LTR retrotransposons in this BAC. In situ hybridisation of the Fatima_2383A24-3 subclone suggests that individual representatives of the Fatima family contribute to the majority of the B-genome specific FISH pattern for BAC_2383A24. Phylogenetic analysis of various Fatima elements available from databases in combination with the data on their insertion dates demonstrated that the Fatima elements fall into several groups. One of these groups, containing Fatima_2383A24-3, is more specific to the B-genome and proliferated around 0.5 2.5 MYA, prior to allopolyploid wheat formation. CONCLUSION: The B-genome specificity of the gypsy-like Fatima, as determined by FISH, is explained to a great degree by the appearance of a genome-specific element within this family for Ae. speltoides. Moreover, its proliferation mainly occurred in this diploid species before it entered into allopolyploidy.Most likely, this scenario of emergence and proliferation of the genome-specific variants of retroelements, mainly in the diploid species, is characteristic of the evolution of all three genomes of hexaploid wheat. PMID- 21635795 TI - Predictors of extra care among magnesium sulphate treated eclamptic patients at Muhimbili National Hospital, Tanzania. AB - BACKGROUND: The inclusion of Magnesium Sulphate (MgSO4) as a gold standard in the treatment of eclampsia has substantially reduced incidences of repeated fits, eclamptic morbidity and deaths. However, despite treatment with MgSO4, a proportion of patients need extra medical/nursing attention and prolonged stay in the intensive care unit (ICU). The literature on the underlying factors for the need of extra care in the MgSO4 era is lacking. This study sought to establish predictors of extra care in ICU among eclamptic patients after treatment with MgSO4 at Muhimbili National Hospital (MNH). METHODS: Data were obtained from hospital records of eclamptic patients who were admitted at MNH and treated with MgSO4 from January 1st to December 31st, 2008. Based on set criteria, patients who needed extra care were identified. Analysis was performed using PASW statistics 18 whereby frequencies, cross-tabulations, bivariate and multiple logistic regressions were performed. RESULTS: A total of 366 eclamptic patients were admitted and treated with MgSO4 at MNH during a 12 month study period in 2008. Most of these (76%) were referred from district hospitals and 132 (36%) met the criteria for extra care in ICU. After adjusting for other variables, the risk of extra care in ICU for patients who were admitted with altered consciousness was double (OR = 2.3; 95% CI: 1.3-4.0) that of the ones admitted in alert state. The risk or need of extra care increased by increasing time to delivery and was doubled (OR = 2.0; 95% CI:1.1-3.7) if it was between 12 and 24 hours and tenfold elevated (OR = 10.0; 95% CI:4.3-23.6) if beyond 24 hours as compared to when time to delivery was less than 12 hours.Abdominal delivery was also independently associated with increased risk compared to vaginal delivery (OR = 2.5; 95%CI: 1.4 4.5). The type of referral and number of fits were associated with extra care in ICU but this association was wholly explained by the clinical status of the patient on admission to MNH and prolonged time lag to delivery. CONCLUSION: We concluded that even with MgSO4 used as the gold standard in the treatment of eclampsia, effective pre-referral care and expedited delivery were crucial in minimizing the need for extra care in ICU. PMID- 21635796 TI - Following autophagy step by step. AB - Autophagy is an evolutionarily conserved lysosomal degradation route for soluble components of the cytosol and organelles. There is great interest in identifying compounds that modulate autophagy because they may have applications in the treatment of major diseases including cancer and neurodegenerative disease. Hundeshagen and colleagues describe this month in BMC Biology a screening assay based on flow cytometry that makes it possible to track distinct steps in the autophagic process and thereby identify novel modulators of autophagy. PMID- 21635797 TI - Trauma care system in Iran. AB - OBJECTIVE: The high burden of injuries in Iran necessitates the establishment of a comprehensive trauma care system. The purpose of this paper is to describe the current status of trauma system regarding the components and function. METHODS: The current status of trauma system in all components of a trauma system was described through expert panels and semi-structured interviews with trauma specialists and policy makers. RESULTS: Currently, various organizations are involved in prevention, management and rehabilitation of injuries, but an integrative system approach to trauma is rather deficient. There has been ongoing progress in areas of public education through media, traffic regulation reinforcement, hospital care and prehospital services. Meanwhile, there are gaps regarding financing, legislations and education of high risk groups. The issues on education and training standards of the front line medical team and continuing education and evaluation are yet to be addressed. Trauma registry has been piloted in some provinces, but as it needs the well-developed infrastructure (regarding staff, maintenance, financial resources), it is not yet established in our system of trauma care. CONCLUSIONS: It seems that one of the problems with trauma care in Iran is lack of coordination among trauma system organizations. Although the clinical management of trauma patients has improved in our country in the recent decade, decreasing the burden of injuries necessitates an organized approach to prevention and management of trauma in the context of a trauma system. PMID- 21635798 TI - One stage anterior-posterior approach for traumatic atlantoaxial instability combined with subaxial cervical spinal cord injury. AB - OBJECTIVES: To explore the clinical features of traumatic atlantoaxial instability combined with subaxial cervical spinal cord injury (CSCI), and to analyze the feasibility, indication and therapeutic effects of anterior-posterior approach in such cases. METHODS: From March 2004 to September 2009, 16 cases with this trauma were admitted and surgically treated in our department. Before surgery, skull traction was performed. Posterior atlantoaxial pedicle screw internal fixation and bone graft fusion were conducted to manage traumatic atlantoaxial instability. As for subaxial CSCI, anterior cervical corpectomy or discectomy decompression, bone grafting and internal fixation with steel plates were applied. RESULTS: All operations were successful. The average operation time was 3 hours and operative blood loss 400 ml. Satisfactory reduction of both the upper and lower cervical spine and complete decompression were achieved. All patients were followed up for 12 to 36 months. Their clinical symptoms were improved by various levels. The Japanese Orthopaedic Association (JOA) scores ranged from 10 to 16 one year postoperatively, 13.95+/-2.06 on average(improvement rate equal to 70.10%). X-rays, spiral CT and MRI confirmed normal cervical alignments, complete decompression and fine implants'position. There was no breakage or loosening of screws, nor exodus of titanium mesh or implanted bone blocks. The grafted bone achieved fusion 3-6 months postoperatively and no atlantoaxial instability was observed. CONCLUSIONS: Traumatic atlantoaxial instability may combine with subaxial CSCI, misdiagnosis of which should be especially alerted and avoided. For severe cases, one stage anterior-posterior approach to decompress the upper and lower cervical spine, together with reposition, bone grafting and fusion, as well as internal fixation can immediately restore the normal alignments and stability of the cervical spine and effectively improve the spinal nervous function, thus being an ideal approach. PMID- 21635799 TI - Nonunion of coronal shear fracture of femoral condyle. AB - Isolated coronal fractures of femoral condyle are rare in adults and nonunion of Hoffa fracture is reported only a few times in the literature. We analyzed six cases of nonunion of Hoffa fractures over a period of three years. Three patients were treated conservatively and three patients had fixation failures. Delay of presentation was 2 months to one year. Treatment protocol consisted of open reduction, excision of pseudoarthrosis, bone grafting and internal fixation along with knee arthrolysis. Union was achieved in all patients at mean 16 weeks. The treatment of nonunion of Hoffa fractures requires careful preoperative planning and meticulous surgical technique. The literature regarding the controversies in fracture management and surgical technique are reviewed. PMID- 21635800 TI - Anatomical and biomechanical study on the interosseous membrane of the cadaveric forearm. AB - OBJECTIVE: To study the anatomical and biomechanical features of the interosseous membrane (IOM) of the cadaveric forearm. METHODS: Ten radius-IOM-ulna structures were harvested from fresh-frozen cadavers to measure the length, width and thickness of the tendinous portion of IOM. Then, the tendinous portion was isolated along with the ulnar and radial ends to which the tendon attached after measurement. The proximal portion of the radius and the distal portion of the ulna were embedded and fixed in the dental base acrylic resin powder. The embedded specimen was clamped and fixed by the MTS 858 test machine using a 10 000 N load cell for the entire tensile test. IOM was stretched at a speed of 50 mm/min until it was ruptured. The load-displacement curve was depicted with a computer and the maximum load and stiffness were recorded at the same time. RESULTS: The IOM of the forearm was composed of three portions: central tendinous tissue, membranous tissue and dorsal affiliated oblique cord. IOM was stretched at a neutral position, and flexed at pronation and supination positions. The tendinous portion of IOM was lacerated in 6 specimens when the point of the maximum load reached to 1021.50 N+/-250.13 N, the stiffness to 138.24 N/m+/-24.29 N/m, and the length of stretch to 9.77 mm+/-1.77 mm. Fracture occurred at the fixed end of the ulna before laceration of the tendinous portion in 4 specimens when the maximum load was 744.40 N+/-109.85 N, the stiffness was 151.17 N/m+/ 30.68 N/m, and the length of the stretch was 6.51 mm+/-0.51 mm. CONCLUSIONS: The IOM of the forearm is a structure having ligamentous characteristics between the radius and the ulna. It is very important for maintenance of the longitudinal stability of the forearm. The anatomical and biomechanical data can be used as an objective criterion for evaluating the reconstructive method of IOM of the forearm. PMID- 21635801 TI - Evaluation of emergency tourniquets for prehospital use in China. AB - OBJECTIVE: Massive hemorrhage is life-threatening during armed conflicts. Tourniquets are important medical devices used to reduce severe bleeding in trauma. The aim of this study was to empirically evaluate the current tourniquets used in China and provide information to emergency nurses in selecting the appropriate tourniquet. METHODS: Five tourniquets were self-applied by 20 healthy participants. The blood flow distal to the tourniquet site was assessed using vascular Doppler ultrasound. Application time, pain, numbness, and other parameters were evaluated. RESULTS: The bladder tourniquet and windlass tourniquet effectively occluded arterial blood flow with success rates higher than 75% in both the upper and lower extremities. The Cargo-strap was the fastest to apply, taking (7.22+/-2.30) s for the upper extremity and (6.48+/-2.40) s for the lower extremity. The rubber tube was the most painful, and the improvised tourniquet was the least efficient. The success rates were higher in the lower extremity than in the upper extremity (P less than 0.05, X(2) equal to 5.714). CONCLUSIONS: The bladder tourniquet and the windlass tourniquet are efficient tourniquets, although the windlass is superior with respect to portability and pain. The Cargo-strap and rubber tourniquets have several disadvantages that reduce their suitability for field use. The improvised tourniquet is not recommended because of low efficiency and severe pain during implementation. PMID- 21635802 TI - Evaluating clinical abdominal scoring system in predicting the necessity of laparotomy in blunt abdominal trauma. AB - OBJECTIVES: Trauma is among the leading causes of death. Medical management of blunt abdominal trauma (BAT) relies on judging patients for whom laparotomy is mandatory. This study aimed to determine BAT patients'signs, as well as paraclinical data, and to clarify the accuracy, sensitivity, specificity, positive and negative predictive value of clinical abdominal scoring system (CASS), a new scoring system based on clinical signs, in predicting whether a BAT patient needs laparotomy or not. METHODS: Totally 400 patients suspected of BAT that arrived at the emergency department of two university hospitals in Tehran from March 20, 2007 to March 19, 2009 were included in this study. They were evaluated for age, sex, type of trauma, systolic blood pressure, Glasgow coma scale (GCS), pulse rate, time of presentation after trauma, abdominal clinical findings, respiratory rate, temperature, hemoglobin (Hb) concentration, focused abdominal sonography in trauma (FAST) and CASS. RESULTS: Our measurements showed that CASS had an accuracy of 94%, sensitivity of 100%, specificity of 88%, positive predictive value of 90% and negative predictive value of 100% in determining the necessity of laparotomy in BAT patients. Moreover, in our analysis, systolic blood pressure, GCS, pulse rate, Hb concentration, time of presentation after trauma, abdominal clinical findings and FAST were also shown to be helpful in confirming the need for laparotomy (P less than 0.05). CONCLUSION: CASS is a promising scoring system in rapid detection of the need for laparotomy as well as in minimizing auxiliary expense for further evaluation in BAT patients, thus to promote the cost-benefit ratio and accuracy of diagnosis. PMID- 21635803 TI - Outcomes of surgical management of tracheobronchial injuries---a case series from a developing country. AB - OBJECTIVE: Tracheobronchial injuries are defined as injuries involving the trachea and/or bronchi from the level of the cricoid cartilage extending up to the division of the bronchi. We present a case series with most of the tracheobronchial injuries found to be sustained after penetrating trauma. METHODS: A retrospective review was performed at the Aga Khan University, Karachi, Pakistan. From January 2004 to December 2009, 168 patients with thoracic trauma were treated, of whom 15 were recognized to have major tracheobronchial and pulmonary injuries. RESULTS: The average age was 31 years with most of the patients being male (14:1). Among them,11 patients had penetrating trauma as the main cause of injury, 3 patients had blunt trauma from road traffic accidents, only 1 patient had combined trauma (blunt and penetrating trauma). Eight patients were diagnosed based on radiological findings. All the patients were treated surgically. Lobectomy was the most common intervention performed in 7 patients. The mortality rate was 7% (1 patient). Most patients survived with no sequelae (10 patients) while 5 survived with disability. We found that penetrating trauma was the leading cause of injury in our series. The severity of injury depends upon the weapon causing the trauma. Patients in our series had multiple injuries and required surgical management. CONCLUSIONS: Tracheobronchial injuries are rare but potentially life threatening. They require quick diagnosis and management. Diagnosis tends to be difficult since there are no specialised diagnostic modalities available at present. PMID- 21635804 TI - Prevalence of seat belt and mobile phone use and road accident injuries amongst college students in Kerman, Iran. AB - OBJECTIVE: To investigate the prevalence of mobile phone and seat belt use in driving in college students aged 18-24 years. METHODS: The study consisted of two phases. In the first phase, a questionnaire was given to 265 college students. The sample contained 188 males and 77 females. In the second phase, the data pertaining to road accident injuries from March 20, 2007 to March 20, 2008 were collected from Bahonar Central Hospital in Kerman, Iran, and analyzed. RESULTS: There were 14.0% of male and 19.0% of female participants traveled belted while driving on urban roads. It indicated that the participants unbelted or using mobile phone were more involved in accidents in last three years. This study also revealed that 19.0% of male and 4.2% of female drivers considered using mobile phone in driving not hazardous. The highest injury and property damage crash rates were 87 and 137 per 100 000 inhabitants, which occured in male group aged 18-24 years. And 30% of all fatalities were 19-24 years old. CONCLUSIONS: More than 50% of college students traveled unbelted. The females were less involved in road accident injuries. The college students aged 18-24 years had the highest percentages of trauma and head injury. PMID- 21635805 TI - Comparing twist-drill drainage with burr hole drainage for chronic subdural hematoma. AB - OBJECTIVE: The surgical management of chronic subdural hematoma (CSDH) is still a controversial issue, and a standard therapy has not been established because of the unclear pathogenic mechanisms in CSDH. The intention of this paper is to find a simple and efficient surgical procedure for CSDH. METHODS: A retrospective study of 448 patients with CSDH by surgical treatment during 2005 to 2009 was conducted in order to compare the efficiency between two different primary surgical methods, i.e. twist-drill drainage without irrigation in Group A (n equal to 178) and one burr-hole with irrigation in Group B (n equal to 270). The results were statistically analyzed. RESULTS: The reoperation rates in Group A and Group B were 7.9% and 11.9% respectively. The good outcome rate was 88.8% and 75.5%, the complication was 7.9% and 20.7% in Group A and Group B, respectively. CONCLUSIONS: The burr-hole drainage with irrigation of the hematoma cavity is not beneficial to the outcome and prognosis. Irrigation is not important in the surgical treatment for CSDH. Thus in initial treatment, twist-drill drainage without irrigation of the hematoma cavity is recommended because it is relatively safe, time-saving and cost-effective. PMID- 21635806 TI - Chronic subdural hematoma associated with sylvian arachnoid cyst in juvenile athletes: report of two cases and literature review. AB - The association of chronic subdural hematoma (CSDH) and arachnoid cyst (AC) is uncommon. We reported 2 juvenile athletes with CSDH associated with AC which occurred in their daily sports activities and reviewed the literature. Both of them were treated surgically, with satisfactory outcome. AC is a common predisposing factor in young patients with CSDH. The complication of intracranial bleeding is an indication for surgical management. Though there are still controversies in the treatment of asymptomatic AC, it is the consensus that the patients with AC should avoid violent sports so as to reduce the incidence of intracranial hemorrhage resulted from head injuries. PMID- 21635807 TI - Transient bone resorption following finger replantation: a report of 3 cases. AB - Radiographic changes consisting of alterations in mineral content, osteopaenia or destructive neuropathy that occur following successful finger replantation have already been described. We report our experience about four fingers in three individuals in whom bone changes developed in the first three months postoperatively with complete "restitution ad integrum". Three patients, 21-49 years old (average 36 years) sustained a clean-cut amputation of four fingers. The first patient had an amputation at the base of the middle phalanx of the index finger and the second patient at the base of the proximal phalanx of the ring finger. The third had an amputation at the base of the first metacarpal bone and the proximal phalanx of the small finger in a five finger amputation. In the first case, two dorsal veins and two palmar digital arteries and nerves were repaired. In the second case, one palmar artery and one dorsal vein were reanastomosed. In the third case at the thumb, two dorsal veins and two palmar digital arteries and nerves were reconstructed. At the small finger, one dorsal vein, one palmar digital artery and two digital nerves were reconstructed. Bone fixation was achieved with two and three K-wires or tension-band wiring. Replantation was successful in all cases. Three weeks after replantation, the X rays showed rapid development of osteopaenia in the juxtaarticular region and metaphyses of the bone. These changes were followed by subperiosteal, intracortical and endosteal bone resorption. No further surgical procedures or splintage were needed and hand therapy was not discontinued. At 10-13 weeks (average 12 weeks) postoperatively, the X-rays showed a complete recovery with new periosteal bone formation. We suggest that the radiographic changes after finger replantation are transient, first evident subperiosteally and progressing centrally. They may reflect small-vessel compromise and microinfarction and transient hyperemia secondary to neurovascular damage or to sympathetic progressive recovery. PMID- 21635808 TI - Ipsilateral fracture dislocations of the hip and knee joints with contralateral open fracture of the leg: a rare case and its management principles. AB - This paper discussed the injury mechanism and management of a patient who had concomitant ipsilateral hip and knee dislocations and contralateral open leg fracture. A 32-year-old man presented with ipsilateral fracture-dislocations of the left hip (Pipkin's type IV) and knee (Moore II) joints and contralateral open fracture of the leg bones after a car accident. After emergency resuscitative measures, the hip joint was reduced and Pipkin's fracture was fixed using Ganz approach with lag screws; knee joint was reduced closely and tibial plateau fracture was stabilized with lateral buttress plate and a transarticular spanning fixator. The open fracture on the other leg was debrided and fixed with an external fixator. There was no instability in both joints after fixation when he was examined under anesthesia. The fractures united after 3 months and the patient had no residual instability of hip and knee. There was no clinical or radiological evidence of osteonecrosis in the hip joint after 6 months. At one year follow-up, he had satisfactory functional outcome with almost normal range of motion at both joints. Ipsilateral hip and knee dislocations are rare injuries and more caution is needed for early diagnosis. A timely appropriate intervention can provide good functional outcome to the patient in this situation. PMID- 21635809 TI - Failure of intramedullary femoral nail with segmental breakage of distal locking bolts: a case report and review of the literature. AB - Breakage of locking bolts is an important cause of interlocking nail failure in femoral fractures. It usually occurs in the form of single breakage in one of the distal bolts of the nail or nail breakage around the distal locking hole. Here we report an unusual case of intramedullary femoral nail failure with segmental breakage of both the distal locking bolts. Such a scenario usually complicates further management. We successfully managed this case with exchange nailing without bone grafting. Here we briefly reviewed the literature regarding such an unusual presentation and discussed in detail the possible etiology of such a presentation and the management options when facing such a complex situation. PMID- 21635810 TI - Immunological mechanisms to establish embryo tolerance in early bovine pregnancy. AB - A well-balanced immunological interaction between mother and the semi-allogenic embryo is of particular importance. The objective of the present study was to analyse mechanisms of immune tolerance in bovine pregnancy during peri implantation. Simmental heifers inseminated with either cryopreserved spermatozoa or seminal plasma were killed 12, 15 or 18 days after oestrus. Uteri were flushed for the recovery of conceptuses and the ipsilateral intercaruncular endometrium was sampled for gene expression analysis. Indoleamine 2,3-dioxygenase (IDO) mRNA, coding for the initial enzyme of the kynurenine pathway, was 18-fold (P < 0.001) more abundant in the endometrium of Day 18 pregnant v. non-pregnant animals. Tandem mass spectrometry revealed a decrease of endometrial l-tryptophan (P = 0.0008), but an increase of l-kynurenine concentration (P = 0.005) from Day 12 to Day 18, suggesting increasing IDO activity (P < 0.03). An in vitro coculture model of endometrial cells showed an induction of IDO expression following interferon-tau exposure primarily in stroma cells, which was confirmed by in situ hybridisation localising IDO mRNA mainly in deep stroma cells. Immunohistochemical analysis revealed fewer CD45-positive leucocytes in the zona basalis of pregnant animals. Elevated IDO activity may reduce the presence of leucocytes in the pregnant endometrium, providing a possible mechanism for protecting the semi-allogenic conceptus from maternal rejection. PMID- 21635811 TI - Differential resistance of mammalian sperm chromatin to oxidative stress as assessed by a two-tailed comet assay. AB - Protamines of eutherian species are cysteine-rich molecules that become cross linked by disulfide bonds during epididymal transit, whereas the protamines of most marsupial species lack cysteine residuals. The present study made use of the differences in protamine structure between eutherian and metatherian mammal spermatozoa to examine the comparative resistance of sperm DNA to oxidative damage in three eutherian species (Mus musculus, Homo sapiens, Sus domesticus) and three metatherian species (Vombatus ursinus, Phascolarctos cinereus, Macropus giganteus). Sperm DNA fragmentation of samples exposed to increasing concentrations of hydrogen peroxide was assessed by means of the two-tailed comet assay. The sperm DNA of the marsupial species studied were significantly more sensitive to oxidative stress than the spermatozoa of eutherian species. Such susceptibility is consistent with the lack of disulfide cross-linking in marsupial sperm chromatin and suggests that the oxidation of thiols to disulfides for chromatin condensation during epididymal transit in eutherian mammals is likely to be important in order to provide stability and protect these cells from the genotoxic effects of adverse environments. PMID- 21635812 TI - Cyclic regulation of apoptotic gene expression in the mouse oviduct. AB - The oviduct is a dynamic structure whose function relies upon cyclic changes in the morphology of both ciliated and secretory luminal epithelial cells. Unfortunately, infection of these epithelial cells by sexually transmitted pathogens can lead to pelvic inflammatory disease, ectopic pregnancies and infertility. The disruption of normal, cyclic apoptosis in the oviducal epithelium appears to be a causal factor of oviducal pathology and therefore, these pathways represent a potential target for diagnosis and therapeutic intervention. The objective of this study was to determine the pattern of expression for apoptotic genes in the oviduct of the naturally cycling mouse, generating fundamental information that can be applied to the development of animal models for research and the identification of targets for disease intervention. Whole oviducts were collected from regular cycling mice killed at 1p.m. on each day of the oestrous cycle and the expression of 84 apoptotic genes determined by targeted PCR super-array. Intact and cleaved caspases were then evaluated by western blotting. The expression of mRNA for genes classified as pro apoptotic (Bad, Bak1 and Bok) and anti-apoptotic (Bag3, Bnip2 and Xiap) was regulated by day (P < 0.05). Differences in the temporal expression of several p53-related genes (Trp53bp2, Trp53inp1 and Trp73), those specific to the TNF superfamily (Tnfrsf10 and Tnfsf10b) and one caspase (Casp14) were also observed (P < 0.05). The cleaved forms of Caspases-3, -6 and -12 were all detected throughout the oestrous cycle. These results represent the first pathway-wide analysis of apoptotic gene expression in the murine oviduct. PMID- 21635813 TI - Maternal endometrial oedema may increase perinatal mortality of cloned and transgenic piglets. AB - The perinatal mortality of cloned animals is a well-known problem. In the present retrospective study, we report on mortality of cloned transgenic or non transgenic piglets produced as part of several investigations. Large White (LW) sows (n = 105) received hand-made cloned LW or minipig blastocysts and delivered either spontaneously or after prostaglandin induction followed by either Caesarean section or vaginal birth. The overall pregnancy rate was 62%, with 26% of pregnancies terminating before term. This resulted in 48 deliveries. The terminated pregnancies consisted of 12 abortions that occurred at 35 +/- 2 days gestation and five sows that went to term without returning to heat and then by surgery showed the uterus without fetal content. The gestation length was for sows with LW piglets that delivered by Caesarean section or vaginally was 115.7 +/- 0.3 and 117.6 +/- 0.4 days, respectively. In sows with minipiglets, the gestation length for those delivered by Caesarean section or vaginally 114.4 +/- 0.2 and 115.5 +/- 0.3 days, respectively. Of the 34 sows that delivered vaginally, 28 gave birth after induction, whereas 6 farrowed spontaneously. Of the 14 sows that delivered after Caesarean section and in the five empty sows, the endometrium and placenta showed severe oedema. Piglet mortality following vaginal delivery was higher than after Caesarean section (31% v. 10%, respectively; P < 0.001). When vaginal delivery occurred spontaneously, the stillborn rate was greater than after induced delivery (56% v. 24%, respectively; P < 0.0001). Internal organ weights were recorded for seven cloned LW piglets and six normal piglets. The relative weight of the heart, liver, kidneys and small intestine was found to be reduced in the cloned piglets (P < 0.05). The present study demonstrates extensive endometrial oedema in sows pregnant with cloned and transgenic piglets, as well as in empty recipients, at term. The growth of certain organs in some of the cloned piglets was reduced and the rate of stillborn piglets was greater in cloned and transgenic piglets delivered vaginally, possibly because of oedema of the fetal-maternal interface. PMID- 21635814 TI - Identification of a beta1 integrin isoform with restricted tissue expression in a teleost fish. AB - The composition and organisation of extracellular matrix (ECM)-related molecules change during development. These components interact with different cell surface receptors to modulate the transduction of signals for cell growth, differentiation, migration, proliferation and apoptosis. Previous findings in the teleost fish gilthead seabream (Sparus aurata L., Teleostei), a marine protandrous hermaphrodite fish, showed that endocrine and immune stimuli are able to modulate the expression of ECM-related molecules, as well as specific correlations between them. In the present study, quantitative reverse transcription-polymerase chain reaction was used to examine the gene expression profile of beta(1) integrin isoform b (ITGB1b) and its possible role in reproductive physiology, especially in relation to spermatogenesis. Expression profiles were analysed in the context of the reproductive cycle (RC) and in relation with other ECM-related molecules, including matrix metalloproteinase (MMP)-2, MMP-9, MMP-13, tissue-specific inhibitor of metalloproteinase (TIMP)-2a, TIMP-2b, collagen (COL1A1) and ITGB1a. Expression of ITGB1b was found in the testis and brain and, to some extent, in endothelial cells. In contrast, ITGB1a was expressed ubiquitously. In the testis, the ITGB1b expression peaked during spermatogenesis, whereas the expression of the other ECM-related molecules is induced mainly during the post-spawning stage, both stages of marked tissue remodelling during the first and second RC in males. In addition, in fish exposed to the endocrine disruptor 17alpha-ethynyloestradiol (at 5 and 50 MUg g(-1) food during 7, 14 and 21 days), ITGB1b expression in the testis was inhibited in a dose- and time-dependent manner and was related to reduced serum levels of testosterone. Together, these results suggest a different functionality for the two ITGB1 isoforms in the gilthead seabream, where ITGB1b is more specifically involved in reproduction. This is the first report of an ITGB1 gene isoform whose expression is restricted to endocrine-related tissues in vertebrates. PMID- 21635815 TI - Loss of the pregnancy-induced rise in cortisol concentrations in the ewe impairs the fetal insulin-like growth factor axis. AB - Maternal cortisol levels increase during pregnancy. Although this change is important for optimal fetal growth, the mechanisms of the changes in growth remain unclear. The hypothesis examined was that alterations in maternal plasma cortisol concentrations are associated with changes in the fetal insulin-like growth factor (IGF) axis. Pregnant ewes in late gestation (115 +/- 0.4 days) were studied: six control animals, five ewes given 1 mg kg(-1) day(-1) cortisol (high cortisol) and five adrenalectomised ewes given 0.5-0.6 mg kg(-1) day(-1) cortisol (low cortisol). Blood samples were taken throughout the experiment and at necropsy (130 +/- 0.2 days) and fetal liver was frozen for mRNA analysis. Fetal IGF-I and insulin plasma concentrations were lower and insulin-like growth factor binding protein-1 (IGFBP-1) concentrations were higher in the low cortisol group compared with those in the control group (P < 0.05). Fetal liver IGF-II and IGFBP 3 mRNA were decreased in low cortisol animals compared with controls (P < 0.05). There were no significant changes in these parameters in the high cortisol group, and there were no changes in fetal liver IGF-I, growth hormone receptor, IGF-I receptor, IGF-II receptor, IGFBP-1 or IGFBP-2 mRNA levels between the groups. These data suggest that reduced fetal IGF availability contributes to reduced fetal growth when maternal cortisol secretion is impaired, but not during exposure to moderate increases in cortisol. PMID- 21635816 TI - Fluctuations in total antioxidant capacity, catalase activity and hydrogen peroxide levels of follicular fluid during bovine folliculogenesis. AB - Follicular fluid is an important environment for oocyte development, yet current knowledge regarding its in vivo oxidant and antioxidant levels remains limited. Examining follicular fluid oxidants and antioxidants will improve understanding of their changes in vivo and contribute to optimisation of in vitro maturation conditions. The aim of the present study was to consider selected markers, namely catalase (CAT) enzyme activity, total antioxidant capacity (TAC) and hydrogen peroxide (H(2)O(2)) in follicular fluid samples (n = 503) originating from bovine antral follicles. The dynamic changes in two relevant antioxidant measures and one reactive oxygen species (ROS) were measured through stages of bovine follicular development and the oestrous cycle. CAT activity and H(2)O(2) levels decreased significantly as follicle size increased, whereas TAC increased significantly as follicle size increased. Lower TAC and higher H(2)O(2) in small follicles suggest increased ROS in the initial stages of folliculogenesis. Because CAT levels are highest in the follicular fluid of small follicles in the setting of an overall low TAC, CAT may represent a dominant antioxidant defence in the initial stages of folliculogenesis. Future studies must focus on other reactive oxygen species and their various scavenger types during antral folliculogenesis. PMID- 21635817 TI - Boar semen can tolerate rapid cooling rates prior to freezing. AB - Two experiments were performed in the present study that demonstrated that boar spermatozoa are capable of surviving rapid cooling rates within a range of 15-5 degrees C before freezing. Boar ejaculates diluted in Beltsville thawing solution (BTS) (1:1, v/v) were held at 17-20 degrees C and shipped over a 24-h time period from two AI centres to a cryobiology laboratory, where they were pooled (Experiment 1) or cryopreserved individually (Experiment 2) using a standard 0.5 mL straw freezing protocol. The effects of cooling before freezing were assessed after thawing through the objective evaluation of sperm motility and flow cytometric analysis of membrane integrity, acrosomal status, changes in membrane lipid architecture monitored by merocyanine and annexin V binding and intracellular production of reactive oxygen species. In Experiment 1 (six replicates), two semen pools (five ejaculates per pool) were cooled from 15 to 5 degrees C at rates of 0.08, 0.13, 0.40 and 1.50 degrees C min(-1). These cooling rates did not result in any significant differences (P>0.05) in any of the post thaw sperm assessments, even in thawed samples incubated under capacitation conditions. In Experiment 2, three individual ejaculates from 16 boars were slowly (0.08 degrees C min(-1)) or rapidly (1.5 degrees C min(-1)) cooled before freezing. A consistent interboar variability (P<0.01) was detected, which was independent of the cooling rate used. Cooling rate only significantly influenced (P<0.05) sperm assessments in four of 16 boars, which exhibited slightly higher percentages of motile cells and intact plasma and acrosomal membranes in the samples that had been cooled slowly. These findings demonstrate that boar spermatozoa undergoing cryopreservation can withstand rapid cooling rates before freezing. PMID- 21635818 TI - Mitochondrial inhibition during preimplantation embryogenesis shifts the transcriptional profile of fetal mouse brain. AB - Environmental stress results in perturbations to mitochondrial function in the preimplantation embryo and hinders subsequent embryo and possibly offspring development. Global gene expression in fetal mouse brain was investigated following targeted mitochondrial inhibition by amino-oxyacetate (AOA) from the 2 cell to the blastocyst stage. Blastocysts were transferred to pseudopregnant recipients and RNA extracted from Day 18 fetal brains for microarray interrogation. Exposure to 5 MUM AOA during preimplantation embryo development induced differential expression of 166 genes (>1.25 fold) in the fetal brain, relative to control medium-cultured embryos. Altered expression pathways included carbohydrate metabolism, neurological development, cellular proliferation and death, DNA replication, recombination and repair. Of 28 genes exhibiting the greatest change in expression, qPCR confirmed that 16 were significantly altered. Targeted qPCR assessment of a further 20 genes associated with methylation, acetylation and mitochondrial dysfunction revealed that three were significantly altered (Immp1l, Nars2, Sat2) and Dmap1 exhibited a sex-specific response to AOA exposure. Only 2/48 genes had significantly altered expression by qPCR (Nola3, Timm8b) in fetal brains exposed to 50 MUM AOA embryo culture, excluding an AOA dose-dependent response. It was concluded that perturbation of mitochondrial function induced by 5 MUM AOA during preimplantation embryo development alters gene expression in the neonatal brain in a manner that suggests that proper brain development may be compromised. PMID- 21635819 TI - Seasonal changes in morphology and steroid receptor expression in the prostate of the brushtail possum (Trichosurus vulpecula): an animal model for the study of prostate growth? AB - The prostate of the brushtail possum undergoes growth and regression during the year. The present study investigated the morphological changes and expression of androgen and oestrogen receptors during the breeding and non-breeding seasons. Prostate tissue was collected from adult possums at 2-monthly intervals. The periurethral and outer glandular areas were separated and the volume of stromal, epithelial and luminal tissues measured in each area. Immunohistochemistry was used to investigate cell proliferation with proliferating cell nuclear antigen (PCNA) and to localise androgen receptor (AR) and oestrogen receptors alpha and beta (ERalpha, ERbeta). Seasonal changes in expression of the three receptors were investigated using quantitative PCR and western blot analysis. During the breeding season the volume of stromal tissue in the periurethral area and the luminal volume in the glandular area significantly increased. The change in periurethral volume was associated with increased PCNA-immunopositive cells. While the localisation of AR to the stromal and epithelial cells did not change, there was a significant increase in receptor expression before the main breeding season. ERalpha and ERbeta expression and localisation did not alter during the year. Similarities in receptor expression and localisation suggest that the possum may be a suitable animal model for the study of human prostate growth. PMID- 21635820 TI - Leukaemia inhibitory factor mediated proliferation of HTR-8/SVneo trophoblast cells is dependent on activation of extracellular signal-regulated kinase 1/2. AB - Leukaemia inhibitory factor (LIF) is one of the cytokines that is indispensable for embryo implantation. The aim of the present study was to investigate the role of activation of extracellular signal-regulated kinase (ERK) 1/2 in LIF-mediated proliferation of HTR-8/SVneo cells. Stimulation of HTR-8/SVneo cells with LIF (50 ng mL(-1)) resulted in an increase in cell proliferation (P < 0.05) via increased transition of cells to the G(2)/M phase of cell cycle. Stimulation with LIF resulted in the activation of both signal transducer and activator of transcription (STAT) 3 Tyr(705) and ERK1/2, but inhibition of ERK1/2 signalling by pretreatment of cells with U0126 (10 uM) for 2h resulted in abrogation of LIF mediated increases in G(2)/M transition, with a significant decrease (P < 0.05) in absolute cell numbers compared with control. Although STAT3 silencing had no effect on LIF-dependent proliferation of HTR-8/SVneo cells, it did result in an increase in cell apoptosis, which increased further upon inhibition of ERK1/2 activation irrespective of LIF stimulation. Stimulation of cells with LIF increased the Bcl-2/Bax ratio, whereas ERK1/2 inhibition decreased the Bcl-2/Bax ratio, even after LIF stimulation. Hence, it can be inferred that ERK1/2 activation is essential for LIF-mediated increases in proliferation and that both STAT3 and ERK1/2 activation are important for the survival of HTR-8/SVneo cells. PMID- 21635821 TI - Regulation of aromatase expression by 1alpha,25(OH)2 vitamin D3 in rat testicular cells. AB - It is well known that the vitamin D endocrine system is involved in physiological and biochemical events in numerous tissues, especially gut, bone and kidney but also testis. Therefore, in this study the effect and mechanisms of action of 1alpha,25(OH)(2) vitamin D(3) (1,25D) on aromatase gene expression in immature rat Sertoli cells were evaluated. Vitamin D receptor transcripts were present in immature Sertoli cells as well as in adult testicular germ cells and somatic cells. The treatment of immature Sertoli cells with 100 nM 1,25D increased the amount of aromatase transcript, mainly in 30-day-old rats. The protein kinase A (PKA) blocker, H89, partially inhibited the 1,25D effect. The stimulation of aromatase gene expression in 30-day-old Sertoli cells by the agonist 1alpha,25(OH)(2) lumisterol(3), and the suppression of the 1,25D effect by the antagonists 1beta,25(OH)(2) vitamin D(3) and (23S)-25-dehydro-1alpha (OH)-vitamin D(3)-26,23-lactone suggested, besides a genomic effect of 1,25D, the existence of non-genomic activation of the membrane-bound vitamin D receptor involving the PKA pathway. PMID- 21635823 TI - Multiple minute digitate hyperkeratoses: a case report. AB - Multiple minute digitate hyperkeratoses (MMDH) is a rare familial or acquired cutaneous eruption of filiform keratoses, typically across the trunk and extremities. Histopathology, distribution, and history can distinguish it from other digitate keratoses. We describe a case of MMDH and discuss it in relation to other disseminated digitate keratoses. PMID- 21635824 TI - Unilateral nevoid telangiectasia syndrome: a case report and review of the literature. AB - We report on a 43-year-old Caucasian female who presented with bright red macules in a unilateral distribution in the left C5-8 and L3-5 dermatomes. Histopathologic examination showed superficial papillary dermal telangiectasia with minimal chronic inflammation. Immunohistochemical stains for estrogen and progesterone receptors (ER/PR) were negative. A diagnosis of unilateral nevoid telangiectasia syndrome (UNTS) was given. UNTS is an uncommon disorder first described by Alfred Blaschko in 1899. It is comprised of telangiectasias occurring in a predominantly unilateral dermatomal distribution and often affecting the trigeminal, cervical, and upper thoracic dermatomes. It can be either congenital or acquired and has a 2:1 female:male ratio. UNTS has been reported in relation to hyperestrogenic states, with half of the reported cases related to pregnancy, puberty, or liver disease. However, the vast majority of cases show no increase in estrogen and progesterone receptors in lesional skin. UNTS may be more common than previously believed, and shows some response to vascular laser therapy. Differential diagnoses include hemangioma, angioma serpiginosum, and rarely, nevus flammeus. PMID- 21635825 TI - Generalized linear porokeratosis: a rare entity with excellent response to acitretin. AB - Linear porokeratosis is a rare disorder of keratinization that usually presents at birth. We report a 17-year-old male with generalized linear porokeratosis, a very rare variant of porokeratosis, with extensive involvement of the trunk and extremities along with nail and genital involvement. The patient was treated with oral acitretin with excellent clinical response. PMID- 21635826 TI - Oral melanosis after tyrosine kinase inhibition with Imatinib for chronic myelogenous leukemia: report of a case and review of the literature. AB - Dermatologic manifestations from therapy with imatinib are well known and frequently include hypopigmentation, and less commonly, hyperpigmentation. There have been few reports of oral hyperpigmentation. We present a case of palatal melanosis related to imatinib therapy for chronic myelogenous leukemia. This case is reported to add to the sparse literature concerning mucosal reactions related to this medication. PMID- 21635827 TI - Cutaneous fusariosis developing in a post-irradiation site. AB - Cutaneous fusariosis is an opportunistic mycosis in immunocompromised patients. We present a novel variation of an immunocompromised patient who developed fusariosis in a previously irradiated site. Irradiation led to atrophy, contraction, fibrosis, barrier disruption, and an altered dermal environment in which the infection developed. Significantly, this is the first case report of fusariosis in a previously irradiated site of an immunocompromised patient. Treatment included debridement and voriconazole. PMID- 21635828 TI - Bilateral segmental neurofibromatosis diagnosed during pregnancy. AB - Bilateral segmental neurofibromatosis is a rare subtype of neurofibromatosis type 1 defined by lesions affecting a single segment of the body and crossing the midline, with no systemic involvement. We present a case diagnosed during pregnancy because of the characteristic increase in size of the lesions during this period. PMID- 21635829 TI - Granuloma annulare of the palms. AB - Granuloma annulare (GA) is a common, benign skin condition, which was first described over a century ago, but still remains an enigma with respect to etiology, associated systemic diseases, and treatment. A number of clinical variants have been classified. We report an atypical presentation of GA localized to the palms. PMID- 21635830 TI - Multifocal tuberculosis revealed by multiple cutaneous gummas in the immunocompetent. AB - Cutaneous tuberculosis is rare. Its occurrence in multifocal tuberculosis (MT) is uncommon and happens frequently in the context of immunosuppression. We report the case of MT with multiple cutaneous gummas and bone and lung involvement that occurred in an apparently immunocompetent patient. PMID- 21635831 TI - A child with giant cell tumor of tendon sheath. AB - A giant cell tumor of tendon sheath (GCTTS) is a soft tissue tumor consisting principally of a proliferation of synovial cells arising from a tendon sheath. GCTTS is the second most common tumor of the hand in general and a majority of GCTTS cases are in patients between 20 and 50 years of age, whereas pediatric cases of GCTTS are uncommon. This report presents the case of a nine-year-old girl with GCTTS arising on her right index finger. PMID- 21635832 TI - A case of verrucous psoriasis. AB - Psoriasis is a common inflammatory disease with a range of clinical presentations and a chronic relapsing course. There have been few reports of verrucous psoriasis, which is distinguished by its warty appearance and characteristic histology. We report a case of this rare form of psoriasis in an elderly woman with a history of plaque-type psoriasis and and discuss the histopathologic findings. PMID- 21635833 TI - Unknown: Yellow plaques on the chest and arms present for more than 6 years in an 80-year-old male. Diffuse normolipemic plane xanthomas presenting in association with myelodysplastic syndrome. AB - Yellow plaques and papules on the skin are hallmarks of xanthomas. These are not always associated with abnormalities in lipid profiles or their associated proteins. In cases of diffuse normolipemic plane xanthomas these cutaneous findings are often associated with myelodyscrasias including monoclonal gammopathy of unknown significance and multiple myeloma. Such clinical presentations may indicate that an evaluation for myelodyscrasias is warranted. PMID- 21635834 TI - Unknown: Papules on the knees. Elastosis perforans serpiginosa (EPS). AB - Elastosis perforans serpiginosa (EPS) can occur in patients with an underlying connective tissue disorder. Down syndrome, osteogenesis imperfecta, acrogeria, Ehlers-Danlos syndrome type IV, Marfan syndrome, pseudoxanthoma elasticum, Rothmund-Thomson syndrome, and scleroderma have all been associated with EPS. The clinical appearance exhibits umbilicated papules arranged in a typical serpiginous pattern, usually located around the neck. Less frequently, lesions may be seen on the face, abdomen, and extremities with a symmetrical distribution, as in our case. The clinical course may be variable with a spontaneous resolution in a few months or years, but frequently new lesions develop leading to persistence and progression of the disorder. We describe a 12 year-old girl affected by Down syndrome who presented a localized form of EPS, which involved only the extremities. Strangely, it resolved spontaneously after biopsy with no recurrence, without therapy. PMID- 21635835 TI - Occupational contact allergy to cephalosporins. AB - Contact sensitivity to systemically administered drugs occurs mainly among healthcare workers and is frequently caused by antibiotics. A 32-year-old nurse presented with a 11/2 year history of hand dermatitis and a 2 month history of palpebral eczema, which were clearly work related. Patch tests with standard and gloves series were negative. Testing of the products commonly handled by the patient were positive for cefradine and cefazolin. Testing of other cephalosporins the patient had not come in contact with were also positive for cefuroxime, ceftriaxone, and cefotaxime. This cross-reactivity may be explained by similar molecular structures. The patient stopped preparing cephalosporin solutions for systemic administration and the lesions cleared. PMID- 21635836 TI - Severe sorafenib-induced hand-foot skin reaction. AB - Sorafenib is a new drug, multikinase inhibitor, which has been recently approved for the treatment of metastatic renal cell carcinoma and hepatocellular carcinoma. Up to 90 percent of patients receiving this drug have been reported to develop dermatological symptoms. Recently, it has been suggested that the appearance of skin toxicity during therapy may indicate antitumor activity. We report a new case of sorafenib-induced severe hand-foot skin reaction, which hindered the patient's normal life. The reaction was successfully treated with topical costicosteroids and discontinuation of sorafenib. However, the patient died one month later. PMID- 21635837 TI - Journal of cosmetic science. PMID- 21635838 TI - Fiber orientation measurement using polarization imaging. AB - In this paper, we present a new technique to determine the orientation of hair fiber, a key parameter in the evaluation of visual appearance of hair. Using polarization imaging and image analysis tools, we are able to measure the orientation of hair fiber for each pixel in the image. A theoretical analysis of the optical set-up is presented. Experimental data on a single fiber, hair tress, and complete head are given. Application to shampoo and conditioner is also demonstrated. PMID- 21635839 TI - Linking enhanced deposition agent functionality with aesthetic performance. AB - This study examines the cationic polymers 1) guar hydroxypropyltrimonium chloride polymers (GHPTC), 2) acrylamidopropyltrimonium chloride/acrylamide copolymer (APTAC/Acm), 3) polyquaternium polymers (PQ-10, PQ-7, PQ-67), and 4) a new polymer system approach for their a) deposition efficiency (as measured by quantifying oils deposited on virgin hair) and b) ability to deliver good wet and dry lubricity to the hair from a cleansing formulation as measured by comb energy and friction characteristics of the hair samples. Conditioning polymer technology approaches 1) acrylamidopropyltrimonium chloride/acrylamide copolymer, 2) a guar hydroxypropyltrimonium chloride polymer, and 3) the new polymer system approach deliver superior deposition of natural conditioning oils and dimethicone materials from anionic/amphoteric surfactant cleansing formulations. These new polymer technologies offer formulators the ability to improve uniformity of deposition as well as deposition efficiency of conditioning agents onto hair, and target the desired hair lubricity. PMID- 21635840 TI - Void and pore formation inside the hair cortex by a denaturation and super contraction process occurring during hair setting with hot irons. AB - An analysis of hair fibers from donors that frequently use hot irons for hair straightening showed the presence of multiple pores and voids (phi approximately 0.1-1.5 MUm) that extend from the cuticle sheath to regions inside the hair cortex. Pore formation in the cortex was found to be confined at its periphery and could be reproduced in the laboratory with virgin hair fibers after the application of various hot-iron straightening cycles. The appearance of pores and voids in the cortex was found to be associated to the production of hot water vapor while the fiber is undergoing mechanical elongation or contraction. The number of pores was seen to rapidly increase with temperature in the range from 190 to 220 degrees C and also with the number of straightening cycles. Larger hair voids (phi approximately 2-5 MUm) were also detected in the cortex. The small pores found at the cortex periphery appear to occur by the simultaneous occurrence of rearrangement of hair proteins, fiber mechanical contraction/expansion, and the flow of super-heated steam. Hot irons create, thus, the conditions for the onset of pore formation as the high temperatures produce superheated steam and soften the native state of hair proteins by a process involving denaturation and changes in the crystalline regions. PMID- 21635841 TI - Graying of the human hair follicle. AB - Quality of life in our society depends crucially on healthy aging, a hallmark of which is the graying hair follicle. During anagen melanocyte precursors migrate to the hair bulb to form the pigmentary unit where they mature and synthesize melanin. Melanin is transferred to the hair shaft forming keratinocytes giving the hair its colour. Graying is the process in which distinct mechanisms lead to deterioration of the hair follicle melanocyte population. We briefly review the hair graying process and state that the aging hair follicle is a valid model for tissue specific aging and a promising target to test therapeutic intervention. PMID- 21635842 TI - Study of hair surface energy and conditioning. AB - A new test method has been developed to determine surface energy of hair fibers through measurements of contact angles at two hair/liquid interfaces. By measuring changes in surface energy of the same hair fiber before and after a cosmetic treatment, effects of active ingredients and the performance of tested formulations can be evaluated.The establishment of the method is based on Fowkes theory (1,2) described with two components, a dispersive and a non-dispersive component. The non-polar liquid used in this study was diiodomethane, and the polar liquid was benzyl alcohol. A Kruss 100 Tensiometer was used to measure contact angles of hair fibers. Virgin dark brown and regular bleached hairs were treated with selected conditioner formulations. Reductions in combing forces of hair tresses before and after respective treatments were correlated with decreases in average surface energy of hair fibers obtained from the corresponding tresses.Experimental results indicate that the average surface energy of hair fibers treated with conditioners decreases and the hydrophobicity of the hair surface increases, the results correlate well with the reduction in combing forces after respective treatments. This research work provides a new methodology to evaluate/screen conditioning performance of hair care ingredients and formulations for development of better products. PMID- 21635843 TI - Thermal styling: efficacy, convenience, damage tradeoffs. AB - We introduce a simple method to explore the efficacy of thermal styling, By using a temperature gradient curling iron we rapidly explore a range of thermal treatment conditions. The thermodynamic literature on the glass transition in keratin fibers explains the surprisingly limited role of elevated temperature in improvements in the efficacy of holding the styled curvature of the fibers. The onset of damage, however, is strongly temperature dependent. This combination of measurements of damage and efficacy shows the range of conditions over which thermal protection products must be functional. PMID- 21635844 TI - Characterization of hair styling formulations targeted to specific multicultural needs. AB - The ethnic hair care market is large and diverse, with many unmet needs, especially when the definition of ethnic varies as much as the hair does. By examining the variety of hair care raw materials now available, we designed hair styling formulations for targeted benefits such as anti-frizz, conditioning, style control, humidity resistance, UV protection and color loss protection. We have characterized three distinctive hair styling formulations targeted to specific multicultural needs. This has been completed by using standard personal care laboratory evaluations including the Diastron Limited (TM)Miniature Tensile Tester for stiffness, the Bossa Nova Technologies (TM) Shine Instrument, high humidity curl control, UV exposure, and expert panel evaluations; the results were substantiated using current state-of-the-art analytical tools, including atomic force microscopy and scanning electron microscopy. Results demonstrate that a varied product portfolio is required for such a diverse market segment. Styling products ranging from alcoholic sprays, leave on styling creams or gels and styling curl activators offer performance attributes that can be utilized on a variety of hair types such as Asian, African, Caucasian and Brazilian. PMID- 21635845 TI - Use of statistical modeling to predict the effect of formulation composition on coacervation, silicone deposition, and conditioning sensory performance of cationic cassia polymers. AB - Formulation composition has a dramatic influence on coacervate formation in conditioning shampoo. The purpose of this study is to correlate the amount of coacervate formation of novel cationic cassia polymers to the corresponding conditioning profiles on European brown hair using silicone deposition, cationic polymer deposition and sensory evaluation. A design of experiments was conducted by varying the levels of three surfactants (sodium lauryl ether sulfate, sodium lauryl sulfate, and cocamidopropyl betaine) in formulations containing cationic cassia polymers of different cationic charge density (1.7 and 3.0m Eq/g). The results show formulation composition dramatically affects physical properties, coacervation, silicone deposition, cationic polymer deposition and hair sensory attributes. Particularly, three parameters are of importance in determining silicone deposition: polymer charge, surfactant (micelle) charge and total amount of surfactant (micelle aspect ratio). Both sensory panel testing and silicone deposition results can be predicted with a high confidence level using statistical models that incorporate these parameters. PMID- 21635846 TI - A study of the distribution of polymer/surfactant coacervate between solution and foam in archetypal shampoo systems. AB - The research reported here attempted to answer the question, "is the foam important in delivering coacervates from shampoos." In order to answer this question, we have measured the amount of polymer in the foam and in the liquid phases of several cationic polymer/anionic surfactant systems by gravimetry and by FTIR techniques. In all cases studied, we discovered that the concentration of solids and, especially the polymer, in the liquid phase and in the foam phase were essentially the same. We conclude that the foam is unlikely to be an important factor in the topical delivery of polymer/surfactant coacervates. PMID- 21635847 TI - Spectrofluorescent characterization of changes in hair chemistry induced by environmental stresses. AB - Hair is frequently exposed to environmental stresses and chemical insults that result in damage to its internal structure and its outer cuticular components. Spectrofluorescence is a useful tool to monitor the health of biological tissues as it can measure the level of tryptophan (Trp), which is representative of protein integrity. In addition to Trp fluorescence, several other fluorophores are also present in hair and are believed to be attributed to kynurenenine, N formylkynurenine, and 3-hydroxykynurenine, which are known metabolic and degradation products of Trp that are affected by environmental stresses normally experienced by hair. In this work, we were able to construct an endogenous fingerprint of fluorescent compounds present in hair by employing a range of excitation wavelengths from 270 nm to 450 nm with a resolution of 2 nm. As a result, we generated surface plots of fluorescence emission as a function of excitation and emission wavelengths (excitation-emission matrices). Thus, we were able to profile the levels of various structural molecules in hair before and after exposure to UV irradiation and thermal straightening irons as well as to chemical treatment such as bleaching and straightening. PMID- 21635848 TI - Hair breakage index: an alternative tool for damage assessment of human hair. AB - Improper hair care, mechanical abrasion, sun damage and chemical treatment changes the physical and morphological characteristics of hair. Several methods involving microscopic techniques, protein loss and assessment of tensile properties of the hair are generally used to evaluate the extent of damage caused. These are also used to determine the protective effect of hair care products. In the present investigation, the hair breakage index (HBI) was used as an alternative tool to determine the change in the properties of hair on weathering. HBI is a measure of the diameter of hair in a given cross sectional area of a marked region of hair on the scalp. The hair diameter changes as we progress towards the tip of the hair due to breakage. The ratio of the diameter of hair bundle in the distal region to the diameter of hair bundle in the proximal region from the scalp is used as an indicator of hair breakage. Higher HBI value is an indicator of hair damage.A study was conducted for duration of 16 weeks to assess the effect of weathering due to grooming practices on HBI values. The HBI and break stress for a group of 30 subjects were measured at baseline and at the end of 16 weeks (NU). Since Coconut oil (CNO) is known to have a positive benefit on tensile properties of hair, another matched group of 30 subjects who oiled their hair daily with CNO was used as a positive control (CNO). The HBI and break stress for this group were also measured at the baseline and after 16 weeks. It was observed that the HBI significantly increased in the NU group versus the CNO user group. The break stress also significantly decreased in the NU group suggesting its correlation with the HBI data. This study demonstrates the usefulness of HBI as a simple and effective tool for determining hair damage and its protection by different hair care products. PMID- 21635849 TI - Anionic/cationic complexes in hair care. AB - The formulation of cosmetic products is always more complicated than studying the individual components in aqueous solution. This is because there are numerous interactions between the components, which make the formulation truly more than the sum of the parts. This article will look at interactions between anionic and cationic surfactants and offer insights into how to use these interactions advantageously in making formulations. PMID- 21635850 TI - Proteomic analysis of hair shaft and nail plate. AB - The protein components of living cells in the hair follicle are amenable to study by standard molecular biological techniques, but identifying those in the hair shaft has been problematic until recently. Most of the protein, primarily keratins and keratin associated proteins, can be extracted under denaturing conditions, but 15-20% is intractable due to transglutaminase-mediated cross linking. Shotgun proteomics now permits identifying >300 constituents of the isopeptide cross-linked proteome and even certain post-translational modifications. The proteins originate from all the intracellular compartments, indicating that the cross-linking process makes effective use of available resources to produce structures with great mechanical stability. Knowing this proteome provides a foundation for correlating defects in hair shaft structure with protein deficiencies. Such investigations can be extended to mouse models of aberrant pelage hair. Thus, inbred mouse strains can be distinguished by their hair proteomes, raising the possibility of similar variation in the human population. The nail plate is also amenable to this shotgun proteomic approach. Providing discrete and noninvasive sampling of the human proteome, these epidermal appendages could have diagnostic utility for certain disease states. PMID- 21635851 TI - Prevention of hair surface aging. AB - The hydrophobic character of the surface of human hair is particularly attributed to the lipid components of the epicuticle and to a layer of covalently bound fatty acids. This outer f-layer mainly consists of 18-methyl eicosanoic acid (18 MEA), which is covalently bound to the underlying protein matrix, forming the epicuticle as composite surface structure. Daily weathering and chemical treatments, specifically oxidative bleaching, decrease the hydrophobicity of the outer hair surface drastically.Multiple daily stress, simulated by an automatic test device including shampooing, blow drying and sun light exposure, changed the lipid composition of hair significantly. A marked loss of 18-MEA was observed. Decreasing contact angles are the direct consequence. A new method to determine the "pseudo-static" contact angle on hair was developed. The results correlate with the corresponding data obtained by dynamic contact angle measurements according to Wilhelmy. Besides that, the resorption time of water droplets by the hair surface provides additional information about the intactness of the outer f layer.Specific proteolipids, which are lipid-modified keratins, are able to reconstruct the surface layer of damaged hair by creating renewed surface hydrophobicity and extending the water resorption time by the hair surface. PMID- 21635852 TI - Polymer/surfactant interactions and nanostructures: current development for cleansing, release, and deposition of actives. AB - Nature exhibits a variety of remarkable phenomena that are useful but difficult to be imitated in real life. Examples are a "touch me not" plant folding up upon being attacked or microbes depositing on ocean vessels even under hostile conditions. Understanding of mechanisms governing these phenomena can prove powerful for developing new classes of cosmetic products. Systems based on polymer/surfactant colloid chemistry are being developed for achieving transport and release of cosmetic and pharmaceutical molecules at desired rates and desired sites. Modifications of the surfactants and polymers provide cleansing properties such as scavenging of odor and sebaceous body excretions and controlled delivery and deposition of sensory/hygienic attributes. New surfactants (sugar based and bio surfactants), hybrid polymers (silicone based and hydrophobically modified) and nanogels have been recently synthesized which may have applications in fields of cosmetics/fragrances/drugs etc. Due to the associative nature of the hydrophobic groups, hybrid polymers can form intramolecular nanodomains at all concentrations of the polymer and inter-molecular aggregates at high concentrations. New hybrid polymers and nano-gel particles can be developed with ability to extract and deliver actives by varying such properties as swelling/shrinking capacity and sensitivity to temperature, shear and dilution. Control of such properties as size, shape and cross linking of nanohybrid particles offer maximum opportunity for producing families of nanovehicles in personal and homecare industry. This review article provides an insight into current developments in field of nano-surfactant science, comprising discussions on nanogel particles, hybrid polymer and liposomes. PMID- 21635853 TI - From conditioning shampoo to nanomechanics and haptics of human hair. AB - Shampoo treatment and hair conditioning have a direct impact on our wellbeing via properties like combability and haptic perception of hair. Therefore, systematic investigations leading to quality improvement of hair care products are of major interest. The aim of our work is a better understanding of complex testing and the correlation with quantitative parameters. The motivation for the development of physical testing methods for hair feel relates to the fact that an ingredient supplier like BASF can only find new, so far not yet toxicologically approved chemistries for hair cosmetics, if an in-vitro method exists.In this work, the effects of different shampoo treatments with conditioning polymers are investigated. The employed physical test method, dry friction measurements and AFM observe friction phenomena on a macroscopic as well as on a nanoscale directly on hair. They are an approach to complement sensoric evaluation with an objective in-vitro method. PMID- 21635854 TI - The effect of various cosmetic pretreatments on protecting hair from thermal damage by hot flat ironing. AB - Hot flat irons are used to create straight hair styles. As these devices operate at temperatures over 200 degrees C they can cause significant damage to hair keratin. In this study, hair thermal damage and the effect of various polymeric pretreatments were investigated using FTIR imaging spectroscopy, DSC, dynamic vapor sorption (DVS), AFM, SEM, and thermal image analysis. FTIR imaging spectroscopy of hair cross sections provides spatially resolved molecular information such as protein distribution and structure. This approach was used to monitor thermally induced modification of hair protein, including the conversion of alpha-helix to beta-sheet and protein degradation. DSC measurements of thermally treated hair also demonstrated degradation of hair keratin. DVS of thermally treated hair shows the reduced water regain and lower water retention, compared to the non-thermally treated hair, which might be attributed to the protein conformation changes due to heat damage. The protection of native protein structure associated with selected polymer pretreatments leads to improved moisture restoration and water retention of hair. This contributes to heat control on repeated hot flat ironing. Thermally stressing hair led to significantly increased hair breakage when subjected to combing. These studies indicate that hair breakage can be reduced significantly when hair is pretreated with selected polymers such as VP/acrylates/lauryl methacrylate copolymer, polyquaternium-55, and a polyelectrolyte complex of PVM/MA copolymer and polyquaternium-28. In addition, polymeric pretreatments provide thermal protection against thermal degradation of keratin in the cortex as well as hair surface damage. The morphological improvement in cuticle integrity and smoothness with the polymer pretreatment plays an important role in their anti-breakage effect. Insights into structure-property relationships necessary to provide thermal protection to hair are presented. PMID- 21635855 TI - [Level of evidence for therapeutic drug monitoring of fluconazole]. AB - Fluconazole is a recommended treatment option for the treatment of invasive candidiasis. Fluconazole is active against most pathogenic Candida spp. except C. kruzei and C. glabrata. Fluconazole is orally well absorbed. Its volume of distribution is reported to be 0.7-1.0 L/kg and it is weakly bound to plasma proteins. The majority of fluconazole dose is excreted via the kidneys. Usually, fluconazole pharmacokinetics display moderate inter-individual variability. However, in presence of renal dysfunction or in pediatrics patients, fluconazole pharmacokinetics might be difficult to predict. Concentration-efficacy relationships are demonstrated whereas concentration-toxicity relationships are sparse. As a consequence, therapeutic drug monitoring is not routinely recommended but can be useful in certain situations. PMID- 21635856 TI - [Level of evidence for therapeutic drug monitoring of itraconazole]. AB - Itraconazole is a triazole antifungal agent that is active against Aspergillus, histoplasmosis, and rare fungal infections. Itraconazole exhibit marked variability in drug concentration as a result of inconsistent absorption, metabolism, or interaction with concomitant medications. Preclinical and clinical data have exhibited a relationship between serum concentrations and treatment efficacy or toxicity, thus therapeutic drug monitoring (TDM) of itraconazole is largely used to optimise therapy. The analysis of bibliographic data demonstrate that, even if the utility of itraconazole's TDM has not been proved by randomized controlled trial or pharmaco-economics studies, it could be useful for managing an absence of response or a drug-drug interactions, or interpreting an adverse effect. However, the interest of this monitoring was proved only in some populations of patients (neutropenics or AIDS patients) so its level of proof varies between levels "potentially useful" and "recommended". PMID- 21635857 TI - [Level of evidence for therapeutic drug monitoring of voriconazole]. AB - Voriconazole is a major antifungal drug with activity against endemic fungi, Candida and Aspergillus species in immunocompromised patients. Voriconazole has a good bioavailability, an high protein binding percentage in plasma and is metabolized in liver via CYP2C19. It presents important neuro- and hepatotoxicities. Some studies determined trough concentrations of voriconazole in plasma using liquid chromatography coupled with UV or tandem mass detection. These studies showed a relationship between trough concentrations of voriconazole and efficacy or toxicity. Indeed, some studies reported a relationship between a lack of clinical response and concentrations below 1 or 2 MUg/mL according to the localization of infection, while toxicities are frequently observed at concentrations above 5 MUg/mL. Some particular populations will have to be taken into account such as children, patients with hemodialysis-dependent renal deficiency or hepatic insufficiency, cystic fibrosis patients or those treated concomitantly with interfering drugs. According to our survey, therapeutic drug monitoring of voriconazole appears recommended. However, controlled studies are still necessary to validate it prospectively and to evaluate pharmacokinetically based methods proposed for individual dose adjustment. PMID- 21635858 TI - [Level of evidence for therapeutic drug monitoring of posaconazole]. AB - Posaconazole, systemic antifungal marketed in France since 2006, is indicated as second line in curative treatment of invasive fungal infections (IFI) (aspergillosis...) and prophylaxis of IFI in patients receiving chemotherapy or hematopoietic stem cell transplantation. The analysis of the literature indicates a concentration-efficacy relationship, but to date, no study has been able to show a concentration-toxicity correlation due to its favourable safety profile and the difficulty to obtain high concentrations. In curative, maintenance of trough plasma concentrations between 0.5 and 1.5 mg/L seems to be associate with an efficacy. In prophylaxis, a threshold of 0.5 mg/L corresponds to a minimal exposure. However this target is not yet well defined. Saturation of absorption above the 800 mg oral dose limits the adjustment of concentrations. As such, the Therapeutic Drug Monitoring of posaconazole can be recommended. PMID- 21635859 TI - [Aripiprazole use in children and adolescent psychiatric patients]. AB - Aripiprazole inaugurates a new generation of antipsychotics called dopamine serotonin system stabilizers. Its mechanism of action is different as aripiprazole is a partial dopamine D(2) and serotonin 5-HT(1A) receptor agonist and 5-HT(2A) receptor antagonist. Therefore, aripiprazole is thought to have an antagonistic action in the mesolimbic pathway but an agonistic action in the mesocortical pathway, tending to normalize the dopaminergic transmission regardless of the type of imbalance. Clinical trials involving children and adolescents have demonstrated the efficacy of aripiprazole in bipolar disorders, schizophrenia, mood disorders associated with pervasive developmental disorders, in tics and Tourette's. The most frequent side effects are extrapyramidal symptoms and sleepiness and are dose-dependent. Nevertheless, contrary to other second-generation antipsychotics available in France, it induces little weight gain, does not modify lipid and glucidic profiles, does not increase prolactin levels, or induce QTc lengthening. The main advantage of aripiprazole is its good safety profile, with different toxicity targets to other second-generation antipsychotics available in France. Aripiprazole appears to be an alternative for children and adolescents who are vulnerable to these side effects and are having trouble coping with them. PMID- 21635860 TI - [Family pharmacy: survey of a sample of patients in Midi-Pyrenees]. AB - OBJECTIVE: Possession of drugs at home in the family pharmacy and self-medication are at risk. METHOD: Appraisal based upon an analysis of 247 questionnaires completed by patients and 116 questionnaires completed by general practitioners in the French department of Haute Garonne (Southwestern, France). RESULTS: Two hundred and forty-four patients were involved in the study. In 80% of cases, women were in charge of family pharmacy who was located in 66% of cases in a unsecurise room and could be reached by children in 17% of cases. Drugs most frequently found: antiseptics (97%), paracetamol (91%), anti-inflammatory drugs (68%), anti-diarrhea (60%). For the physicians 52 useable questionnaires, 80% of physician were confronted with one of three risks: self-medication, drug autolysis, poisoning in children. CONCLUSION: Women are the referent of the family pharmacy. The doctors seem best placed to a message of prevention through minimal advice. PMID- 21635861 TI - Pulmonary arterial hypertension and benfluorex: 5 case reports. AB - Benfluorex, an amphetamine derivative, was marketed as an adjunctive drug for patients with hypertriglyceridemia or diabetes with overweight. We describe here 5 cases of pulmonary arterial hypertension (PAH) associated to a treatment with this drug and notified to the Midi-Pyrenees Pharmacovigilance Centre. All cases are women, exposed to benfluorex during at least 3 years. In most of cases, benfluorex was used off-licence, for overweight. PMID- 21635862 TI - [Drug induced hyponatremia. About a series of 54 cases notified to the regional center of pharmacovigilance of Saint-Etienne]. AB - OBJECTIVE: Analyze the most frequently reported drug in iatrogenic hyponatremia, explaining the mechanism in question. METHODS: Retrospective study on 54 notified cases between the Jan.1(st) 2003 and Dec. 31(st) 2009. The analysis focuses on drug classes, accountability and mechanism; risk factors, severity and the evolution of the case. RESULTS: Fourty-nine cases have been retained, 75,5% of the time it was an inappropriate secretion of antidiuretic hormone (SIADH) involving a selective serotonin reuptake inhibitor (SSRI) in 15 cases, an anticonvulsant in 7 cases, a proton pump inhibitor (PPI) in 7 cases or other drugs in 11. The hypo-osmolar hypovolemic hyponatremia was 12.2% of cases, incriminating a thiazide diuretic once in two. The mechanism was different or indeterminate in 12.2% of cases. In 67% cases were serious. The evolution was often favourable. CONCLUSION: Drug induced hyponatremia can be serious. It must control natremia with patients at risk or symptomatic and conduct the etiological diagnosis. PMID- 21635863 TI - [Sarcoidosis and anti-TNF: a paradoxical class effect? Analysis of the French Pharmacovigilance system database and literature review]. AB - OBJECTIVES: To identify and characterize the observations of sarcoidosis occurring during anti-TNF blockade collected in the French Pharmacovigilance system database and reported in the literature. RESULTS: Seven cases were reported in the French Pharmacovigilance system database and 39 cases (37 original) have been reported internationally. Monoclonal antibodies (infliximab and adalimumab) and fusion protein (etanercept) are equally involved. Sarcoidosis have been confirmed histologically and occurred predominantly in the rheumatoid arthritis (22) and spondylarthropathy (16). CONCLUSION: The lack of protopathic bias suggests that these paradoxical sarcoidosis occurring during treatment with anti-TNF are a class-effect, as with psoriasis, uveitis, and IBD reported under similar conditions. Their pathogenesis remains unclear. PMID- 21635864 TI - [Profile of psychoactive substances consumption in workplace]. AB - PURPOSE: The aim of this study was to identify profiles of psychoactive substances consumers among workers according to their professional characteristics. METHODS: In 2006, 2213 workers participated in "Mode de Vie et Travail" (Drugs and Work) cross-sectional survey. Data were collected by a self administered questionnaire concerning general and professional characteristics and the consumption of psychoactive substances (psychoactive drugs, tobacco, alcohol and cannabis) during the professional medical visit. We identified consumer profiles with a hierarchical ascendant classification as statistical method. RESULTS: We underlined five profiles associated with psychoactive substance consumption: (1) alcohol consumers in the workplace were sales engineers satisfied with their employment, (2) alcohol consumers after their work were not satisfied with their lives, (3) cannabis consumers were men professionally satisfied but suffering from job insecurity, (4) smokers were workers with professional responsibilities under time pressure, and finally (5) poly-consumers had strong professional constraints. CONCLUSIONS: This study guides occupational physicians on psychoactive substances consumption among a worker population. PMID- 21635865 TI - Evaluation of the effects of ethinylestradiol on sexual differentiation in the olvas-GFP/STII-YI medaka (transgenic Oryzias latipes) strain as estimated by proliferative activity of germ cells. AB - We evaluated the effects of 17(-ethinylestradiol (EE(2)) on sexual differentiation in transgenic olvas-GFP/STII-YI medaka (Oryzias latipes) in terms of the proliferative activity of germ cells. This strain contains the green fluorescent protein (GFP) gene fused to the regulatory region of the medaka vasa gene, and germ cell-specific expression of GFP can be visualized in living (transparent) individuals. From 0 days post-hatch (0 dph) onwards, juveniles were exposed to graded concentrations of EE(2) (25.2-1710 ng/L) for 35 days. The gonads of live specimens were monitored by measuring their size and calculating their GFP-fluorescence area. GFP-fluorescent area in control females was about 10 times that in control males at 10 days posthatch (dph) whereas the gonadal size of 10 dph males that had been exposed to 158 ng/L of EE(2) significantly increased up to twice the size of control males, indicating that abnormal sexual differentiation towards female might occur in these individuals. Histological examination and identification of the sex-linked marker SL1 indicated that male to female sex reversal occurred at EE(2) exposure >=45.1 ng/L at 35 dph. These results suggest that observation of proliferative activity of germ cells in the olvas-GFP/STII-YI strain could be applied to facilitated screening fish model to detect adverse effects on sexual differentiation as early as 10 dph juveniles. PMID- 21635866 TI - Potential synergistic effects of microcystins and bacterial lipopolysaccharides on life history traits of Daphnia galeata raised on low and high food levels. AB - Metastudies have found no consistent effects of the cyanobacterial toxin microcystin on Daphnia, and there are discrepancies between field observations and experiments. Confounding factors include absence or presence of alternative high quality food or the presence of bioactive compounds, other than microcystins in cyanobacteria. Of specific interest are lipopolysaccharides (LPS) on the outer cell wall. LPS may have a number of biological effects, including reduced detoxication of microcystins in plants and animals. When grazing seston in the field, filterfeeders take up heterotrophic bacteria attached to cyanobacteria, as well as free-living bacteria. The LPS produced by heterotrophic bacteria have been shown to be much more harmful than cyanobacterial LPS. We performed two experiments in which we tested for potential synergistic effects between bacterial LPS and microcystins. Full-factorial experiments separated the main effects and interactions between (i) food quantity as well as food quality (addition of the green alga Scenedesmus), (ii) presence or absence of strains that vary in amount and composition of microcystins (microcystin free strain NIVA CYA43, moderate microcystin producing strain NIVA-CYA140 and high microcystin producing strain PCC7820), and (iii) presence or absence of bacterial LPS on different life history traits of Daphnia galeata. We measured juvenile growth rate, age and size at first reproduction, death before first reproduction and standard carbon content of Daphnia. From the experiments we conclude that microcystin-producing Microcystis had deleterious effects on the life history of D. galeata, but especially when the availability of high quality green algal food was limited in comparison to the supply of microcystin producing strain PCC7820. In the experiment in which PCC7820 was used as microcystin-producing strain, addition of LPS lowered SCC of Daphnia, but had no effects on other life history parameters. The interaction between Microcystis strain, Microcystis concentration and LPS was highly significant in case of PCC7820, but not in case of CYA-140, indicating that the effects of LPS and its interactions with microcystin on Daphnia life history were strongly context dependent. PMID- 21635867 TI - Atrazine exposure affects growth, body condition and liver health in Xenopus laevis tadpoles. AB - Six studies were performed regarding the effects of atrazine, the most frequently detected pesticide in fresh water in the US, on developing Xenopus laevis tadpoles exposed 5 days post-hatch through Nieuwkoop Faber Stage 62. The levels of atrazine tested included those potentially found in puddles, vernal ponds and runoff soon after application (200 and 400 MUg/L) and a low level studied by a number of other investigators (25 MUg/L). One study tested 0, 25 and 200 MUg/L, another tested 0, 200 and 400 MUg/L, while the remaining four studies tested 0 and 400 MUg/L. During all exposures, mortality, growth, metamorphosis, sex ratio, fat body (a lipid storage organ) size and liver weights, both relative to body weight, were evaluated. In selected studies, feeding behavior was recorded, livers and fat bodies were histologically evaluated, liver glycogen and lipid content were determined by image analysis, and immunohistochemical detection of activated caspase-3 in hepatocytes was performed. The NOEC was 25 MUg/L. None of these exposure levels changed sex ratios nor were intersex gonads noted, however, no definitive histological evaluation of the gonads was performed. Although a marginal increase in mortality at the 200 MUg/L level was noted, this was not statistically significant. Nor was there an increase in mortality at 400 MUg/L versus controls. At the 400 MUg/L level, tadpoles were smaller than controls by 72 h of exposure and remained smaller throughout the entire exposure. Appetite was not decreased at any exposure level. Slowed metamorphosis was noted only at 400 MUg/L in two of five studies. Livers were significantly smaller in the study that tested both 200 and 400 MUg/L, yet no pathological changes or differences in glycogen or lipid stores were noted. However, livers from 400 MUg/L exposed tadpoles had higher numbers of activated caspase-3 immunopositive cells suggesting increased rates of apoptosis. Fat body size decreased significantly after exposure to 200 and 400 MUg/L although these organs still contained some lipid and lacked any pathology. Since this was noted across all studies, it was considered the most sensitive indicator of atrazine exposure measured. The changes noted in body and organ size at 200 and 400 MUg/L atrazine indicated exposure throughout development compromised the tadpoles. Significant reductions in fat body size could potentially decrease their ability to survive the stresses of metamorphosis or reduce reproductive fitness as frogs rely on stored lipids for these processes. PMID- 21635868 TI - The efficacy of simulated solar disinfection (SODIS) against Ascaris, Giardia, Acanthamoeba, Naegleria, Entamoeba and Cryptosporidium. AB - The antimicrobial activity of simulated solar disinfection (SODIS) in the presence and absence of riboflavin against various protozoa and helminth organisms was investigated in this study. Assays were conducted in transparent 12 well microtitre plates containing a suspension of test organisms in the presence or absence of 250 MUM riboflavin. Plates were exposed to simulated sunlight at an optical irradiance of 550 Wm(-2) (watts per square metre) delivered from a SUNTESTTM CPS+ solar simulator. Aliquots of the test suspensions were taken at set time points and the viability of the test organisms was determined by either culture, microscopy or flow cytometry where applicable. With Acanthamoeba, Naegleria, Entamoeba and Giardia exposure to SODIS at an optical irradiance of 550 Wm(-2) for up to 6h resulted in significant inactivation of these organisms. The addition of riboflavin to this system significantly increased the level of inactivation observed with cysts of A. castellanii. With Cryptosporidium oocysts and Ascaris ova exposure to SODIS in the presence and absence of riboflavin for 6 8h resulted in a negligible reduction in viability of both organisms. In this present study we have been able to show that SODIS is effective against a variety of previously untested waterborne organisms and with A. castellanii cysts the addition of micro-molar concentrations of riboflavin can enhance cyst inactivation. However, care must be taken as Ascaris larvae continue to develop inside the ova after exposure to SODIS and Cryptosporidium remain impermeable to propidium iodide staining indicating they may still be infectious. PMID- 21635869 TI - Multilocus molecular and phylogenetic analysis of phlebotomine sand flies (Diptera: Psychodidae) from southern Italy. AB - This study reports a combined analysis of mitochondrial and ribosomal DNA target regions of phlebotomine sand flies (Diptera: Psychodidae) from the Mediterranean region. A ~900 bp long fragment of the mitochondrial DNA encompassing regions within cytb and nd1 gene and the complete ITS2 ribosomal region (~500 bp) were sequenced and characterized for Phlebotomus perniciosus, Phlebotomus perfiliewi, Phlebotomus neglectus, Phlebotomus papatasi, and Sergentomyia minuta, captured in two sites of southern Italy. From one to eight mitochondrial haplotypes and from one to three ITS2 sequence types were found for the examined specimens according to the different sand fly species. The mean interspecific difference in the mitochondrial sequences was of 16.1%, with an overall intraspecific nucleotide variation from 0.1 to 2.8%. A higher interspecific difference (mean 25.1%) was recorded for the ITS2 sequence, with an overall intraspecific nucleotide variation up to 4.9%. The sequence types alignment of ITS2 region showed that all phlebotomine specimens possessed a split 5.8S rRNA, consisting of a mature 5.8S rRNA and a 2S rRNA separated by a short transcribed spacer. Phylogenetic analysis of the Phlebotomus spp. sequences, herein determined and of those available in GenBankTM were concordant in clustering P. neglectus, P. perfiliewi and P. papatasi with the same species collected from different geographic areas of the Mediterranean basin in four main clades for mtDNA and ITS2, respectively. This study demonstrates the utility of multilocus sequencing, provides a dataset for the molecular identification of the most prevalent phlebotomine sand flies in southern Europe and defines the phylogenetic relationships among species examined. PMID- 21635870 TI - Plk1 is negatively regulated by RNF8. AB - RNF8 is a nuclear protein having an N-terminal forkhead-associated (FHA) domain and a C-terminal RING-finger (RF) domain. Depletion of RNF8 caused cell growth inhibition and cell cycle arrest at not only S but also G2/M phases. In addition, cell death was frequently observed in RNF8-depleted cells. Analyses of time-lapse microscopy revealed that the cells died in mitosis and interphase. To elucidate the RNF8 function in M phase, the Plk1 content in RNF8-depleted cells was examined. The amount of RNF8 decreased time-dependently, whereas Plk1 reciprocally increased by transfection of RNF8 siRNA. Protein contents of RNF8 and Plk1 among various cell lines were also compared. RNF8 in normal cell lines was much higher than that in many cancer cell lines. Conversely, Plk1 in normal cell lines was lower than in cancer cell lines. These results suggest that RNF8 is downregulated in many cancer cells and inversely correlated with Plk1. PMID- 21635871 TI - Crystal structure of Pseudomonas aeruginosa PA2196, a putative TetR family transcriptional repressor. AB - PA2196 of Pseudomonas aeruginosa is a putative transcriptional regulator and belongs to the TetR family repressor that is involved in adaptations to environmental changes and bacterial antibiotic resistance. The crystal structure of PA2196 determined to 2.4A resolution revealed nine alpha-helical bundles that can be divided into N-terminal DNA binding domain with an alpha-helix-turn-alpha helix motif and C-terminal ligand binding domain with a hydrophobic ligand binding pocket. The distance between the N-terminal domains of homodimeric PA2196 suggested that our structure is similar to the DNA-bound form of other TetR family proteins. The C-terminal ligand binding pocket is composed mainly of hydrophobic residues and has a volume of about 523A(3) with two openings. PA2196 binds to the upstream region and can regulate the downstream genes that are chemical modification enzymes. Our crystal structure of PA2196 provides insights about the DNA recognition and ligand binding characteristics. PMID- 21635872 TI - Acquired and intrinsic BRAF inhibitor resistance in BRAF V600E mutant melanoma. AB - The discovery of activating BRAF V600E mutations in 50% of all cutaneous melanomas has revolutionized the understanding of melanoma biology and provided new strategies for the therapeutic management of this deadly disease. Highly potent small molecule inhibitors of BRAF are now showing great promise as a novel therapeutic strategy for melanomas harboring activating BRAF V600E mutations and are associated with high levels of response. This commentary article discusses the latest data on the role of mutated BRAF in the development and progression of melanoma as the basis for understanding the mechanism of action of BRAF inhibitors in the preclinical and clinical settings. We further address the issue of BRAF inhibitor resistance and outline the latest insights into the mechanisms of therapeutic escape as well as describing approaches to prevent and abrogate the onset of both intrinsic and acquired drug resistance. It is likely that our evolving understanding of melanoma genetics and signaling will allow for the further personalization of melanoma therapy with the goal of improving clinical responses. PMID- 21635873 TI - Compound C stimulates heme oxygenase-1 gene expression via the Nrf2-ARE pathway to preserve human endothelial cell survival. AB - We recently identified adenosine monophosphate-activated protein kinase (AMPK) as a novel inducer of heme oxygenase-1 (HO-1) and surprisingly found that compound C (6-[4-(2-piperidin-1-yl-ethoxy)-phenyl]-3-pyridin-4-yl-pyrazolo[1,5-a] pyrimidine), a cell-permeable inhibitor of AMPK, could also elevate HO-1 suggesting other AMPK-independent actions for this agent. In this study, we investigated the biochemical mechanism by which compound C stimulates HO-1 expression in human endothelial cells (ECs) and determined the biological significance of the induction of HO-1 by compound C in these cells. Compound C stimulated a concentration- and time-dependent increase in HO-1 expression and an increase in HO-1 promoter activity that was abrogated by mutating the antioxidant responsive elements (AREs) in the HO-1 promoter or by overexpressing a dominant negative mutant of NF-E2-related factor 2 (Nrf2). Compound C also stimulated Nrf2 expression this was associated with an increase in the production of reactive oxygen species and with a decline in intracellular glutathione levels. Interestingly, the glutathione donor N-acetyl-l-cysteine or the NADPH oxidase inhibitor apocynin blocked the induction of HO-1 by compound C. Finally, compound C stimulated EC death and this was potentiated by silencing HO-1 expression and reversed by the administration of CO, biliverdin, or bilirubin. In conclusion, this study demonstrates that compound C stimulates HO-1 gene expression in human vascular endothelium via the activation of the Nrf2/ARE signaling pathway to counteract compound C-mediated cell death. The ability of compound C to induce HO 1 expression may contribute to the pleiotropic actions of this agent and suggest caution when using compound C to probe for AMPK functions. PMID- 21635874 TI - N-acetyl-L-cysteine (NAC) inhibit mucin synthesis and pro-inflammatory mediators in alveolar type II epithelial cells infected with influenza virus A and B and with respiratory syncytial virus (RSV). AB - 64% of chronic obstructive pulmonary disease (COPD) exacerbations are caused by respiratory infections including influenza (strains A and B) and respiratory syncytial virus (RSV). They affect the airway epithelium increasing inflammatory and apoptosis events through mechanisms involving ROS generation, and induce the release of mucins from epithelial cells that are involved in the deterioration of the patient's health during the course of the disease. The antioxidant NAC has proved useful in the management of COPD reducing symptoms, exacerbations and accelerated lung function decline. It has been shown to inhibit influenza virus replication and to diminish the release of inflammatory and apoptotic mediators during virus infection. The main objective of this study is to analyze the effects of NAC in modulating MUC5AC over-expression and release in an in vitro infection model of alveolar type II A549 cells infected with influenza (strains A and B) and RSV. We have also analyzed virus replication and different pro inflammatory responses. Our results indicate a significant induction of MUC5AC, IL8, IL6 and TNF-alpha that is strongly inhibited by NAC at the expression and at the release level. It also decreased the intracellular H(2)O(2) concentration and restored the intracellular total thiol contents. Mechanisms of NAC included inhibition of NF-kappaB translocation to the cellular nucleus and phosphorylation of MAPK p38. NAC also inhibited replication of the three viruses under study. This work supports the use of antioxidants in order to ameliorate the inflammatory effects of different viral infections during COPD exacerbations. PMID- 21635875 TI - Phase diagram of mixed monolayers of stearic acid and dimyristoylphosphatidylcholine. Effect of the acid ionization. AB - The aim of this work is to study the phase diagram of mixed monolayers composed of dimyristoylphosphatidylcholine (DMPC) and stearic acid (SA) at different ionic strength and bulk pH of the aqueous subphase. In this way, the effect of ionization of SA on the interaction and thus on phase separation with the DMPC matrix can be analyzed. To this purpose, we first determined the ionization state of pure SA monolayers as a function of the bulk subphase pH. The SA monolayers are nearly fully ionized at pH 10 and essentially neutral at pH 4 and the mixture of DMPC and SA was studied at those two pHs. We found that the DMPC-enriched phase admits more SA if the SA monolayer is in a liquid-expanded state, which is highly related to the acid ionization state, and thus to the bulk pH and ionic strength. At pH 4 the molecules hardly mix while at pH 10 the mixed monolayer with DMPC can admit between 30 and 100% of SA (depending on the lateral pressure) before phase separation is established. The addition of calcium ions to the subphase has a condensing effect on SA monolayers at all pHs and the solubility of SA in the DMPC matrix does not depend on the bulk pH in these conditions. The observed phase diagrams are independent on the manner in which the state of the mixed film is reached and may thus be considered states of apparent equilibrium. PMID- 21635876 TI - Ether lipids. AB - The naturally occurring 1-O-alkyl-sn-glycerols and their methoxylated congeners, 1-O-(2'-methoxyalkyl)-sn-glycerols, are biologically active compounds, ubiquitously found in nature as diacyl glyceryl ether lipids and phosphoether lipids. The chief objective of this article is to provide a comprehensive and up to date review on such ether lipids. The occurrence and distribution of these compounds in nature are extensively reviewed, their chemical structure and molecular variety, their biosynthesis and chemical synthesis and, finally, their various biological effects are described and discussed. An unprecedented biosynthesis of the 2'-methoxylated alkylglycerols is proposed. The first synthesis of enantiopure (Z)-(2'R)-1-O-(2'-methoxyhexadec-4'-enyl)-sn-glycerol, the most prevalent 2'-methoxylated type alkylglycerol present in cartilaginous fish, is described. It was accomplished by a highly convergent five step process. PMID- 21635877 TI - The relationship between bone turnover markers and BMD decreasing rates in Chinese middle-aged women. AB - BACKGROUND: The relationship between bone turnover markers (BTMs) and BMD decreasing rate (BDR) in Chinese women is unclear. Wu investigated the relationship between (BTMs) and BDR at various skeletal sites in Chinese middle aged women. METHODS: A cross-section study of 555 healthy Chinese women over 35 60years of age. BMD at posteroanterior spine, the left hip, and the left forearm were measured with a DXA. Levels of serum osteocalcin (OC), bone-specific alkaline phosphatase (BAP), cross-linked N-terminal telopeptides of type I collagen (sNTX) and total urinary deoxypyridinoline (uDPD) were determined. RESULTS: BDR at various skeletal sites had significant negative correlation with serum OC(r=-0.395 to -0.530), BAP(r=-0.297 to -0.486), and sNTX(r=-0.207 to 0.272). After adjustment of age and weight, serum OC, BAP, and sNTX rather than total uDPD still exhibited significant correlations with BDR. Stepwise regression analyses showed that, serum OC and BAP were the significantly negative determinants of BDR. Between 4.7-27.7% and 1.2-16.1% of the changes in BDR were determined by serum OC and BAP, respectively. However, sNTX and total uDPD had no significant effect on BDR at various skeletal sites. CONCLUSIONS: This study indicated the correlation between BTMs and early-stage BDR in Chinese middle-aged women and suggested that serum OC and BAP, rather than sNTX and total uDPD, are the key determining factors of early BMD decreases. PMID- 21635878 TI - Human chorionic gonadotropin in pregnancy diagnostics. AB - Human chorionic gonadotropin (hCG) is a 237 aminoacid glycoprotein hormone composed of two dissimilar alpha and beta subunits noncovalently linked by charge interactions, which are both required for the biological activity of the hormone. Due to structural heterogeneity, hCG exists in biological fluids as a mixture of different isoforms, i.e., intact active hormone (hCG), nicked hCG (hCGn), free beta subunits (hCGbeta), free alpha subunit (hCGalpha), beta-core fragment (hCGbetacf, predominantly detected in urine and containing amino acids 6-40 and 55-92 linked by disulphide bridges) and nicked free beta-subunit (hCGbetan). Although the measurement of hCG might be useful in a kaleidoscope of clinical conditions, such as diagnosis, monitoring and follow-up of pregnancy-related disorders, prenatal screening and gynecological cancers, the leading application is still the diagnosis of pregnancy, where it can be measured quantitatively either in serum or urine, in the latter case also using qualitative and rapid immunoassays. Since there is still debate as to whether serum or urine tests are to be preferred for establishing a diagnosis of pregnancy, we discuss here the main analytical and clinical aspects of hCG measurement for the diagnosis of pregnancy, highlighting the advantages and limitations of assessing hCG in urine and serum. PMID- 21635879 TI - Replicated association between genetic variation in the PARK2 gene and blood pressure. AB - BACKGROUND: The PARK2 gene encodes Parkin which is linked to the mitochondrial fusion, fission and mitophagy. In the previous study, single nucleotide polymorphisms (SNPs) in the PARK2 gene have been reported to be associated with blood pressure (BP) in Nigerian families. We aimed to confirm whether the genetic variation of the PARK2 gene influenced the susceptibility to BP and hypertension in Korean population. METHODS: We performed the quantitative BP trait analysis and hypertension case-control analysis for the 227 SNPs in the PARK2 gene in the Korean Association Resource (KARE) cohort (8512 subjects) and the Korean Urban Epidemiology (KUE) cohort (3703 subjects) by the independent association analysis and meta-analysis. RESULTS: Two SNPs, rs9456721 and rs6902041, were significantly associated with systolic BP (SBP) and diastolic BP (DBP), respectively, in the KARE subjects, and their association p-values were below the Bonferroni-corrected significance level. The replication analysis in the KUE subjects revealed a significant association between the SNP, rs6902041 and BP. CONCLUSIONS: These results indicate that genetic variation in the PARK2 gene is significantly associated with BP not only in the Nigerian population but also in the Korean population. This study may provide insight into the genetic basis of hypertension related to the mitochondrial quality control. PMID- 21635880 TI - Serum galectin-3 levels in patients with nonalcoholic fatty liver disease. AB - OBJECTIVES: Galectin-3 might serve as a biomarker of human metabolic alterations. We measured serum levels of galectin-3 in patients with nonalcoholic fatty liver disease (NAFLD) and examined their association with clinical and histological phenotypes. DESIGN AND METHODS: Serum levels of galectin-3 were assayed in 71 patients with biopsy-proven NAFLD and 39 controls. RESULTS: Serum galectin-3 levels did not differ in patients with NAFLD (median 4.1 ng/mL; interquartile range: 1.5-5.5 ng/mL) compared with healthy controls (median 3.1 ng/mL; interquartile range: 0.8-7.5 ng/mL, P=0.93). Among patients with NAFLD, however, serum galectin-3 levels correlated significantly with BMI (r=0.267, P<0.05). This association persisted after adjustment for potential confounders (beta=0.30; t=2.11, P<0.05). CONCLUSIONS: Although galectin-3 was modestly associated with BMI, our results do not support the hypothesis that levels of this molecule are altered in patients with NAFLD. PMID- 21635881 TI - Analytical interference of quinolone antibiotics and quinine derived drugs on urinary protein determined by reagent strips and the pyrogallol red-molybdate protein assay. AB - OBJECTIVES: To investigate the analytical interference of drugs in urinary protein and to estimate the lowest interfering concentrations. DESIGN AND METHODS: Drug supplemented urine samples were compared to the control with two reagent strips and the total protein was determined using Pyrogallol Red Molybdate (PRM). RESULTS: False-positive interferences occurred with Multistix 10 SG for hydroxychloroquine, levofloxacin and ofloxacin. No interference was observed with Combur 10 Test M. Statistically significant false-positive interferences were observed in the PRM assay with all tested drugs, and lowest interfering concentrations were mostly above estimated therapeutic concentrations. The PRM assay "confirmed" the results of the Multistix dipstick, so a real proteinuria could be presumed from the double analytical interference. CONCLUSIONS: This is the first report of analytical interference by quinolone and quinine derivatives in the PRM assay. Special attention to patients using these drugs is needed to minimize errors in the interpretation of urinary protein results. PMID- 21635882 TI - Endocrine profile and phenotype-genotype correlation in unrelated patients with non-classical congenital adrenal hyperplasia. AB - OBJECTIVES: The aim of this study was to identify the molecular defect in a group of 37 unrelated Greek Cypriot patients affected by NC-CAH and evaluate the relationship between the genotype, phenotype and adrenal androgen levels. DESIGN AND METHODS: Clinical evaluation, biochemical analysis of 17-OHP, Testosterone, Androstenedione, DHEA-S, direct DNA sequencing and MLPA analyses. RESULTS: Eleven known mutations were identified with the p.V281L being the most predominant and observed in 68.9% of the alleles. There was no difference between the two genotypes (mild/mild and mild/severe) with clinical presentation, whereas a proportional relationship between the type of mutation and adrenal androgen levels was found. CONCLUSION: The frequency of the underlying genetic defect in our patients with NC-CAH is similar to that observed in most Mediterranean populations. Although the genotype cannot solely explain the clinical expression of NC-CAH, discrimination between mild and severe alleles is crucial in antenatal diagnosis and genetic counselling. PMID- 21635883 TI - Third intracellular loop of glucagon like-peptide-1 receptor is coupled with endogenous mono-ADP-ribosyltransferase - novel type of receptor regulation? AB - Our previous studies revealed the main role of the third intracellular loop (IC(3)) of glucagon-like peptide-1 receptor (GLP-1 receptor), in G-protein activation, where the presence or absence of agonist and the receptor phosphorylation seemed to be the only regulatory mechanisms. In order to further study the signaling mechanisms of GLP-1 receptor, we investigated the effect of the third intracellular loop-derived peptide on endogenous mono-ADP ribosyltransferase mediated mono-ADP-ribosylation of G-proteins beta subunit in CHO cells. Results showed an inhibitory effect of IC(3) peptide on mono-ADP ribosylation of beta subunit, obviously via the mechanism of competitive inhibition. Excluding the activity of this inhibitory mechanism via pertussis toxin-sensitive G proteins, the direct functional coupling of IC(3) of GLP-1 receptor and endogenous mono-ADP-ribosyltransferase was confirmed. We suggest that this arginine specific enzymatic posttranslational modification of third intracellular loop of GLP-1 receptor might represent a possible novel mechanism of receptor activity regulation and the pharmacological potential in treatment of diabetes mellitus type 2. PMID- 21635884 TI - Oral administration of the thrombin receptor antagonist E5555 (atopaxar) attenuates intimal thickening following balloon injury in rats. AB - Thrombin is a powerful agonist for a variety of cellular responses including platelet aggregation and vascular smooth muscle cell (SMC) proliferation. These actions are mediated by a thrombin receptor known as protease-activated receptor 1 (PAR-1). Recently we discovered that 1-(3-tert-butyl-4-methoxy-5 morpholinophenyl)-2-(5,6-diethoxy-7-fluoro-1-imino-1,3-dihydro-2H-isoindol-2 yl)ethanone hydrobromide (E5555, atopaxar) is a potent and selective thrombin receptor antagonist. This study characterized the pharmacological effects of E5555 on SMC proliferation in vitro and in a rat model of intimal thickening after balloon injury in vivo. E5555 selectively inhibited rat aortic SMC proliferation induced by thrombin and thrombin receptor-activating peptide (TRAP) with half maximal inhibitory concentration (IC(50)) values of 0.16 and 0.038 MUM, respectively. E5555 did not inhibit rat SMC proliferation induced by basic fibroblast growth factor (bFGF) and platelet-derived growth factor (PDGF) at concentrations up to 1MUM. In addition, E5555 inhibited human aortic SMC proliferation induced by thrombin at concentrations of 0.3 and 3units/ml with IC(50) values of 0.028 and 0.079 MUM, respectively, whereas it did not affect bFGF-induced proliferation at concentrations up to 1MUM. Repeated oral administration of 30 mg/kg E5555 (once daily for 16 days) significantly reduced neointimal formation in the balloon-injured rat arterial model. These results suggested that a PAR-1 antagonist could be effective for treating restenosis following vascular intervention in addition to preventing thrombus formation. E5555 could thus have therapeutic potential for restenosis and chronic atherothrombotic disease. PMID- 21635885 TI - Protective effect of Etoricoxib against middle cerebral artery occlusion induced transient focal cerebral ischemia in rats. AB - Stroke is the third leading cause of global death and disability. Cyclooxygenase 2 mRNA has been shown to be up-regulated after stroke and also the time window of its expression extends from 4 to 12 h. The objective of this study was to elucidate the protective effect of Etoricoxib (a selective Cyclooxygenase-2 inhibitor) against transient middle cerebral artery occlusion induced behavioral, biochemical and histological alterations. Transient ischemia reperfusion significantly caused behavioral (neurological deficits, decreased locomotor activity and rotarod performance), biochemical (increased lipid peroxidation and nitrite concentration, while decreased superoxide dismutase and catalase activity) and histological (increased infarct volume) changes. Etoricoxib (3 and 10 mg/kg, i.p.) significantly reversed the alterations caused by cerebral ischemia however, 1 mg/kg dose was not found effective in any of the parameters. Finally, we can conclude that Etoricoxib has beneficial effects against transient middle cerebral artery occlusion model in rats. The present study indicates that Etoricoxib may be considered as a potential candidate in the treatment of stroke, clinically. PMID- 21635886 TI - Effect of glutamate receptor antagonists microinjected into the nucleus accumbens on place aversion induced by naloxone in single-dose, morphine-treated rats. AB - It is well established that acute morphine withdrawal can be observed following opioid receptor antagonism in rodents. Glutamate receptor antagonists can attenuate the conditioning place aversion (CPA) induced by naloxone in single dose, morphine-treated rats. Anatomically, the nucleus accumbens appears to be involved in opiate dependence. In the present study, we examined the effects of various glutamate receptor antagonists in the nucleus accumbens on naloxone induced CPA in rats. MK-801 (an NMDA receptor antagonist), GYKI52466 (an AMPA receptor antagonist), and MCPG (a metabotropic glutamate receptor antagonist) significantly attenuated naloxone-induced CPA following microinjection into the accumbens. In contrast, none of the agents showed place conditioning ability on their own in either morphine-exposed or naive rats. The present study suggests that glutamate receptors in the nucleus accumbens play a key role in the motivational component of withdrawal during acute morphine dependence. PMID- 21635887 TI - CIN85 regulates ubiquitination and degradative endosomal sorting of the EGF receptor. AB - CIN85 has been demonstrated to interact with a number of proteins involved in endocytosis and intracellular sorting. However, the exact functional role of CIN85 in endocytosis remains unclear. We have investigated whether CIN85 plays a role in EGF-induced EGF receptor (EGFR) internalization, as previously suggested, or whether CIN85 is rather involved in endosomal sorting of the EGFR. When over expressing a dominant negative interfering CIN85 mutant consisting of three SH3 domains only, we found that internalization of EGF was inhibited. However, when knocking down CIN85 by RNAi, the EGF-EGFR uptake appeared similar to in control cells. Furthermore, in CIN85 depleted cells, EGF-induced ubiquitination of the EGFR was decreased, and degradation of EGF-EGFR complexes was delayed. Our data further demonstrated that depletion of CIN85 increased the recycling of EGF, suggesting that CIN85 plays a role in endosomal sorting of the ubiquitinated EGFR. Our data also demonstrated that CIN85 was constitutively associated with Hrs, and this strengthens the hypothesis of a functional role of CIN85 in endosomal EGFR sorting. PMID- 21635889 TI - Protective effects and mechanism of tetramethylpyrazine against lens opacification induced by sodium selenite in rats. AB - Tetramethylpyrazine (TMP), extracted from the Chinese herbal medicine Ligusticum wallichii franchat (chuan xiong in Chinese), is a potent anti-free radical and calcium antagonist. Correspondingly, two important hypotheses in the causation of cataracts are free radical toxicity and calcium ion overload. In this study we investigated the effect of TMP on lens opacification induced by sodium selenite in rats, addressing the potential of TMP eye drops to prevent and treat cataracts. Results showed that the extent of lens opacification in the untreated Normal Control group (NC group) was significantly less than that of selenite injected untreated rats (MC group) on days 3, 5, 7 and 10 (p < 0.001), while TMP treated selenite-injected rats (TMP group) had less lens opacification than the MC group on days 3, 5, 7 and 10 (p < 0.05). Compared with the NC group, the MC group had significantly decreased activity of super-oxide dismutase (SOD), glutathione peroxidase (GSH-PX) and catalase (CAT) and significantly elevated malondialdehyde (MDA) and calcium ion content (p < 0.001). Compared with the MC group, the activity of (SOD), (GSH-PX) and (CAT) were significantly higher while (MDA) and calcium ion levels were significantly lower in the TMP group at all time points (p < 0.01). The findings demonstrate that the selenite-induced cataract rat models were successfully built and the TMP eye drops can delay lens opacification induced by sodium selenite in rats. The mechanism by which TMP preserves lens transparency from selenite treated animals is associated with the lenses' ability to maintain normal levels of activity of SOD, GSH-PX and CAT and normal concentrations of MDA and calcium ion. PMID- 21635888 TI - The isolated muscle fibre as a model of disuse atrophy: characterization using PhAct, a method to quantify f-actin. AB - Research into muscle atrophy and hypertrophy is hampered by limitations of the available experimental models. Interpretation of in vivo experiments is confounded by the complexity of the environment while in vitro models are subject to the marked disparities between cultured myotubes and the mature myofibres of living tissues. Here we develop a method (PhAct) based on ex vivo maintenance of the isolated myofibre as a model of disuse atrophy, using standard microscopy equipment and widely available analysis software, to measure f-actin content per myofibre and per nucleus over two weeks of ex vivo maintenance. We characterize the 35% per week atrophy of the isolated myofibre in terms of early changes in gene expression and investigate the effects on loss of muscle mass of modulatory agents, including Myostatin and Follistatin. By tracing the incorporation of a nucleotide analogue we show that the observed atrophy is not associated with loss or replacement of myonuclei. Such a completely controlled investigation can be conducted with the myofibres of a single muscle. With this novel method we can distinguish those features and mechanisms of atrophy and hypertrophy that are intrinsic to the muscle fibre from those that include activities of other tissues and systemic agents. PMID- 21635890 TI - A novel free radical scavenger rescues retinal cells in vivo. AB - The benzopyran BP (3,4-dihydro-6-hydroxy-7-methoxy-2,2-dimethyl-1(2H)-benzopyran) is a free radical scavenger that is structurally similar to alpha-tocopherol and has provided neuro-protection in a number of disease models where oxidative stress is a causative factor. A novel derivative of BP with improved lipid solubility, which we have designated BP3, was synthesized and its neuro protective efficacy subsequently analyzed in three mouse models of retinal disease in vivo. In the acute light damage model, balb/c mice received a single intra-peritoneal injection (200 mg/kg) of BP3 one hour prior to phototoxicity, reducing photoreceptor degeneration for up to 48 h post insult. In the rd10/rd10 mouse, a chronic model of inherited retinal degeneration, systemic dosing with BP3 on alternate days between post-natal day 18 and 25 preserved rod photoreceptor numbers and cone photoreceptor morphology. Finally, NMDA induced toxicity in retinal ganglion cells was diminished for at least 72 h after the initial insult by a single dose of BP3. In each disease model, BP3 alleviated cellular oxidative burden as MDA levels were markedly reduced. These results demonstrate that systemically administered BP3 has potent free radical scavenging capacity in the retina and may represent a single therapeutic strategy applicable across several retinopathies. PMID- 21635891 TI - Identification of Drosophila SOD3 and its protective role against phototoxic damage to cells. AB - Superoxide dismutase (SOD) is one of several major proteins that regulate removal of superoxide. Three isoforms of SOD exist in mammals. It has long been thought that Drosophila lacks the SOD3 gene. However, a putative SOD3 gene sequence (dSod3) in the Drosophila genome was reported recently. Thus we investigated whether dSod3 truly functions as a SOD3 homolog in Drosophila. We found that dSod3 not only retains SOD activity but also properties of secreted proteins, as do other SOD3s. In addition, the dSod3 protein alleviates ultraviolet-induced cellular damage. These results suggest that dSod3 functions as an extracellular SOD3. PMID- 21635892 TI - Cellular DBP and E4BP4 proteins are critical for determining the period length of the circadian oscillator. AB - The phenotypes of mice carrying clock gene mutations have been critical to understanding the mammalian clock function. However, behavior does not necessarily reflect cell-autonomous clock phenotypes, because of the hierarchical dominance of the central clock. We performed cell-based siRNA knockdown and cDNA overexpression and monitored rhythm using bioluminescent reporters of clock genes. We found that knockdown of DBP, D-box positive regulator, in our model led to a short-period phenotype, whereas overexpressing of DBP produced a long-period rhythm when compared to controls. Furthermore, knockdown and overexpressing of E4BP4, D-box negative regulator, led to an opposite effect of DBP. Our experiments demonstrated that D-box regulators play a crucial role in determining the period length of Per1 and Per2 promoter-driven circadian rhythms in Rat-1 fibroblasts. PMID- 21635893 TI - Enteric neuronal density contributes to the severity of intestinal inflammation. AB - BACKGROUND & AIMS: Enteric neurons have been reported to be increased in inflamed regions of the bowel in patients with inflammatory bowel disease or intestinal neurogangliomatosis. It is impossible to determine whether this hyperinnervation predates intestinal inflammation, results from it, or contributes to its severity in humans, so we studied this process in mice. METHODS: To determine whether the density of enteric neurons determines the severity of inflammation, we studied transgenic mice that have greater than normal (NSE-noggin mice, which overexpress noggin under the control of the neuron-specific enolase promoter) or fewer than normal (Hand2(+/-) mice) numbers of neurons in the enteric nervous system. Colitis was induced with trinitrobenzene sulfonic acid or dextran sulfate sodium, and the intensity of the resulting inflammation in Hand2(+/-) and NSE-noggin mice was compared with that of wild-type littermates. RESULTS: Severity of each form of colitis (based on survival, symptom, and histologic scores; intestinal expression of genes that encode proinflammatory molecules; and levels of neutrophil elastase and p50 nuclear factor kappaB) were significantly reduced in Hand2(+/-) mice and significantly increased in NSE-noggin animals. Neither mouse differed from wild-type in the severity of delayed-type hypersensitivity (edema, T-cell and neutrophil infiltration, or expression of interleukin-1beta, interferon-gamma, or tumor necrosis factor-alpha) induced in the ears using 2,4 dinitro-1-fluorobenzene. Transgene effects on inflammation were therefore restricted to the gastrointestinal tract. CONCLUSIONS: The severity of intestinal inflammation is associated with the density of the enteric innervation in mice. Abnormalities in development of the enteric nervous system might therefore contribute to the pathogenesis of inflammatory bowel disease. PMID- 21635894 TI - Mechanisms responsible for progesterone's protection against lordosis-inhibiting effects of restraint I. Role of progesterone receptors. AB - Progestins and antiprogestins are widely used therapeutic agents in humans. In many cases, these are indicated for the treatment of reproductive activities. However, progesterone has widespread physiological effects including a reduction of the response to stress. We have reported that 5 min of restraint reduced lordosis behavior of ovariectomized rats hormonally primed with estradiol benzoate. When ovariectomized rats received both estradiol benzoate and progesterone priming, restraint had minimal effects on lordosis. Progesterone influences behavior through classical intracellular progesterone receptor mediated nuclear events as well as extranuclear events. How these multiple events contribute to the response to stress is unclear. The current project was designed to initiate examination of the mechanisms responsible for progesterone's ability to protect against the effects of the restraint. In the first experiment, ovariectomized rats, primed with 10 MUg estradiol benzoate, received 500 MUg progesterone 4 h, 1 h, or 30 min before restraint. When progesterone was injected 4h before restraint, progesterone eliminated the effects of restraint. In contrast, progesterone 30 min before restraint offered no protection. Effects of progesterone 1h before restraint were equivocal allowing the suggestion that less than 4h of progesterone priming might be sufficient. In the second experiment, the synthetic progestin, medroxyprogesterone, was shown to mimic effects of progesterone in preventing effects of restraint. Finally, the progesterone receptor antagonist, RU486, attenuated progesterone's protection against restraint. These findings offer evidence that ligand-activated progesterone receptor mechanisms contribute to the maintenance of lordosis behavior in the presence of mild stress. PMID- 21635895 TI - Pre- and post-weaning cold exposure does not lead to an obese phenotype in adult Brandt's voles (Lasiopodomys brandtii). AB - Evidence has shown that postnatal undernutrition, overnutrition and cold stress are associated with imbalanced metabolic regulation as rodents achieve adulthood. In this study, we used a breeding colony of Brandt's voles (Lasiopodomys brandtii), a wild rodent species from the Inner Mongolia grasslands in China, to examine the effects of pre- and post-weaning cold exposure on the adult body (fat) mass, serum hormones and hypothalamic neuropeptides. Unlike laboratory rodents, vole offspring exposed to pre-weaning cold did not exhibit overweight or obese phenotypes in adulthood compared with unexposed controls. Moreover, adult male voles that remained in colder conditions had less body mass and lower serum leptin levels despite having higher food intake compared to other groups. To understand the mechanism of this unexpected regulation, hypothalamic gene expression was assessed for pre- and post-weaning cold exposure. Voles exposed to cold before weaning increased hypothalamic, orexigenic agouti-related protein (AgRP) and decreased anorexigenic proopiomelanocortin (POMC) mRNA expression at weaning. These expression changes were associated with hyperphagia and catch-up growth after weaning. Interestingly, these changes in hypothalamic neuropeptides were short lasting because in adult voles these differences were no longer apparent, which might explain why the pre-weaning, cold-exposed voles did not become obese in adulthood. These data suggest that some species do not develop an obese phenotype in response to early life cold stress. PMID- 21635896 TI - Resisting resistin; it's good for the heart. PMID- 21635897 TI - Three-dimensional analysis of the association of viral particles with mitochondria during the replication of Rice gall dwarf virus. AB - Examination of cultured insect vector cells that had been infected with Rice gall dwarf virus (RGDV), using transmission electron microscopy and confocal microscopy, revealed the presence of clusters of virus-coated mitochondria around viroplasms in which replication and assembly of RGDV occurred, suggesting a role for mitochondria in supplying the energy required for viral morphogenetic processes. Electron tomography revealed that RGDV particles on the surface of mitochondria are arrayed in an orderly but loose manner, unlike tightly packaged particles in vesicular compartments, suggesting the presence of counterpart molecules on the surface of mitochondria. The viral particles in close proximity to mitochondria were aligned along intermediate filaments, which might serve as scaffolds for the anchorage of these particles. RGDV has a putative mitochondrion targeting sequence on the outer surface of the outer-capsid protein P8. The arrangement of RGDV particles around mitochondria suggests that the region of the P8 protein containing the mitochondrion-targeting sequence might attach to a molecule like a receptor on the outer mitochondrial membrane. Our analysis demonstrates the three-dimensional arrangement and molecular basis for the mitochondrial proximity of RGDV particles during viral replication. PMID- 21635898 TI - Exploring the Structure-Function Loop Adaptability of a (beta/alpha)(8)-Barrel Enzyme through Loop Swapping and Hinge Variability. AB - Evolution of proteins involves sequence changes that are frequently localized at loop regions, revealing their important role in natural evolution. However, the development of strategies to understand and imitate such events constitutes a challenge to design novel enzymes in the laboratory. In this study, we show how to adapt loop swapping as semiautonomous units of functional groups in an enzyme with the (beta/alpha)(8)-barrel and how this functional adaptation can be measured in vivo. To mimic the natural mechanism providing loop variability in antibodies, we developed an overlap PCR strategy. This includes introduction of sequence diversity at two hinge residues, which connect the new loops with the rest of the protein scaffold, and we demonstrate that this is necessary for a successful exploration of functional sequence space. This design allowed us to explore the sequence requirements to functional adaptation of each loop replacement that may not be sampled otherwise. Libraries generated following this strategy were evaluated in terms of their folding competence and their functional proficiency, an observation that was formalized as a Structure-Function Loop Adaptability value. Molecular details about the function and structure of some variants were obtained by enzyme kinetics and circular dichroism. This strategy yields functional variants that retain the original activity at higher frequencies, suggesting a new strategy for protein engineering that incorporates a more divergent sequence exploration beyond that limited to point mutations. We discuss how this approach may provide insights into the mechanism of enzyme evolution and function. PMID- 21635899 TI - Regulation of mammalian cell cycle progression in the regenerating liver. AB - The process of cell division in mammalian cells is orchestrated by cell-cycle dependent oscillations of cyclin protein levels. Cyclin levels are controlled by redundant transcriptional, post-translational and degradation feedback loops. How each of these separate loops contributes to the regulation of the key cell cycle events and to the connection between the G1-S transition and the subsequent mitotic events is under investigation. Here, we present an integrated computational model of the mammalian cell cycle based on the sequential activation of cyclins. We validate the model against experimental data on liver cells (hepatocytes), which undergo one or two rounds of synchronous circadian clock gated cell divisions during liver regeneration, after partial hepatectomy (PH). The model exhibits bandpass filter properties that allow the system to ignore strong but transient, or sustained but weak damages after PH. Bifurcation analysis of the model suggests two different threshold mechanisms for the progression of the cell through mitosis. These results are coherent with the notion that the mitotic exit in mammalian cells is bistable, and suggests that Cdc20 homologue 1 (Cdh1) is an important regulator of mitosis. Regulation by Cdh1 also explains the observed G2/M phase prolongation after hepatocyte growth factor (HGF) stimulation during S phase. PMID- 21635900 TI - Two-intermediate model to characterize the structure of fast-folding proteins. AB - This paper introduces a new model that enables researchers to conduct protein folding simulations. A two-step in silico process is used in the course of structural analysis of a set of fast-folding proteins. The model assumes an early stage (ES) that depends solely on the backbone conformation, as described by its geometrical properties--specifically, by the V-angle between two sequential peptide bond planes (which determines the radius of curvature, also called R radius, according to a second-degree polynomial form). The agreement between the structure under consideration and the assumed model is measured in terms of the magnitude of dispersion of both parameters with respect to idealized values. The second step, called late-stage folding (LS), is based on the "fuzzy oil drop" model, which involves an external hydrophobic force field described by a three dimensional Gauss function. The degree of conformance between the structure under consideration and its idealized model is expressed quantitatively by means of the Kullback-Leibler entropy, which is a measure of disparity between the observed and expected hydrophobicity distributions. A set of proteins, representative of the fast-folding group - specifically, cold shock proteins - is shown to agree with the proposed model. PMID- 21635901 TI - Adaptive compressive learning for prediction of protein-protein interactions from primary sequence. AB - Protein-protein interactions (PPIs) play an important role in biological processes. Although much effort has been devoted to the identification of novel PPIs by integrating experimental biological knowledge, there are still many difficulties because of lacking enough protein structural and functional information. It is highly desired to develop methods based only on amino acid sequences for predicting PPIs. However, sequence-based predictors are often struggling with the high-dimensionality causing over-fitting and high computational complexity problems, as well as the redundancy of sequential feature vectors. In this paper, a novel computational approach based on compressed sensing theory is proposed to predict yeast Saccharomyces cerevisiae PPIs from primary sequence and has achieved promising results. The key advantage of the proposed compressed sensing algorithm is that it can compress the original high-dimensional protein sequential feature vector into a much lower but more condensed space taking the sparsity property of the original signal into account. What makes compressed sensing much more attractive in protein sequence analysis is its compressed signal can be reconstructed from far fewer measurements than what is usually considered necessary in traditional Nyquist sampling theory. Experimental results demonstrate that proposed compressed sensing method is powerful for analyzing noisy biological data and reducing redundancy in feature vectors. The proposed method represents a new strategy of dealing with high dimensional protein discrete model and has great potentiality to be extended to deal with many other complicated biological systems. PMID- 21635902 TI - The clock and wavefront model revisited. AB - The currently accepted interpretation of the clock and wavefront model of somitogenesis is that a posteriorly moving molecular gradient sequentially slows the rate of clock oscillations, resulting in a spatial readout of temporal oscillations. However, while molecular components of the clocks and wavefronts have now been identified in the pre-somitic mesoderm (PSM), there is not yet conclusive evidence demonstrating that the observed molecular wavefronts act to slow clock oscillations. Here we present an alternative formulation of the clock and wavefront model in which oscillator coupling, already known to play a key role in oscillator synchronisation, plays a fundamentally important role in the slowing of oscillations along the anterior-posterior (AP) axis. Our model has three parameters which can be determined, in any given species, by the measurement of three quantities: the clock period in the posterior PSM, somite length and the length of the PSM. A travelling wavefront, which slows oscillations along the AP axis, is an emergent feature of the model. Using the model we predict: (a) the distance between moving stripes of gene expression; (b) the number of moving stripes of gene expression and (c) the oscillator period profile along the AP axis. Predictions regarding the stripe data are verified using existing zebrafish data. We simulate a range of experimental perturbations and demonstrate how the model can be used to unambiguously define a reference frame along the AP axis. Comparing data from zebrafish, chick, mouse and snake, we demonstrate that: (a) variation in patterning profiles is accounted for by a single nondimensional parameter; the ratio of coupling strengths; and (b) the period profile along the AP axis is conserved across species. Thus the model is consistent with the idea that, although the genes involved in pattern propagation in the PSM vary, there is a conserved patterning mechanism across species. PMID- 21635903 TI - Modeling non-homologous end joining. AB - Non-homologous end joining (NHEJ) is an important DNA repair pathway for DNA double-strand breaks. Several proteins, including Ku, DNA-PKcs, Artemis, XRCC4/Ligase IV and XLF, are involved in the NHEJ for the DNA damage detection, DNA free end processing and ligation. The classical model of NHEJ is a sequential model in which DNA-PKcs is first recruited by the Ku bound DNA prior to any other repair proteins. Recent experimental study (McElhinny et al., 2000; Costantini et al., 2007; Mari et al., 2006; Yano and Chen, 2008) suggested that the recruitment ordering is not crucial. In this work, by proposing a mathematical model in terms of biochemical reaction network and performing stability and related analysis, we demonstrate theoretically that if DSB repair pathway independent of DNA-PKcs exists, then the classical sequential model and new two-phase model are essentially indistinguishable in the sense that DSB can be repaired thoroughly in both models when the repair proteins are sufficient. PMID- 21635904 TI - The distribution of alternating AT sequences in eukaryotic genomes suggests a role in homologous chromosome recognition in meiosis. AB - There are general features of chromosome dynamics, such as homologue recognition in early meiosis, which are expected to involve related sequence motifs in non coding DNA, with a similar distribution in different species. A search for such motifs is presented here. It has been carried out with the CONREPP programme. It has been found that short alternating AT sequences (10-20 bases) have a similar distribution in most eukaryotic organisms, with some exceptions related to unique meiotic features. All other microsatellite and repeat sequences vary significantly in different organisms. It is concluded that the unique structural features and uniform distribution of alternating AT sequences indicate that they may facilitate homologous chromosome pairing in the early preleptotene stage of meiosis. They may also play a role in the compaction of DNA in mitotic chromosomes. PMID- 21635905 TI - Fading memory model for airway smooth muscle dynamic response. AB - This work presents the application of a fading memory model to describe the behavior of contracted airway smooth muscle (ASM) for two biophysical cases: finite duration length steps and longitudinal sinusoidal oscillations. The model parameters were initially determined from literature data on transient step length change response and subsequently the model was applied to the two cases. Results were compared with previously published experimental data on ASM oscillations. The model confirms a trend observed in the experimental data which shows that: (i) the value of tissue length change is the most important factor to determine the degree of cross-bridge detachment and (ii) a strong correlation exists between increasing frequency and declining stiffness until a certain frequency (~25 Hz) beyond which frequency dependence is negligible. Although the model was not intended to simulate biophysical events individually, the data could be explained by cross-bridge cycling rates. As the frequency increases, cross-bridge reattachment becomes less likely, until no further cross-bridge attachment is possible. PMID- 21635906 TI - Flavonoids: potential Wnt/beta-catenin signaling modulators in cancer. AB - Flavonoids are polyphenolic compounds found throughout the plant kingdom. They occur in every organ but are usually concentrated in leaves and flowers. During the last two decades, in vitro and in vivo studies demonstrated that flavonoids have inhibitory effects on human diseases through targeting of multiple cellular signaling components. Wnt/beta-catenin signaling regulates proliferation, differentiation and fate specification in developmental stages and controls tissue homeostasis in adult life. For these reasons, this pathway has received great attention in the last years as potential pathway involved in distinct Human pathologies. In this review we discuss the emerging potential mechanisms for flavonoids on Wnt/beta-catenin signaling in cancer and possible investigation strategies to understand flavonoids mode of action on this signaling pathway. PMID- 21635907 TI - Role of the primary motor cortex in L-Dopa-induced dyskinesia and its modulation by 5-HT1A receptor stimulation. AB - While serotonin 5-HT1A receptor (5-HT1AR) agonists reduce L-DOPA-induced dyskinesias (LID) by normalizing activity in the basal ganglia neurocircuitry, recent evidence suggests putative 5-HT1AR within the primary motor cortex (M1) may also contribute. To better characterize this possible mechanism, c-fos immunohistochemistry was first used to determine the effects of systemic administration of the full 5-HT1AR agonist +/-8-OH-DPAT on L-Dopa-induced immediate early gene expression within M1 and the prefrontal cortex (PFC) of rats with unilateral medial forebrain bundle (MFB) dopamine (DA) lesions. Next, in order to determine if direct stimulation of 5-HT1AR within M1 attenuates the onset of LID, rats with MFB lesions were tested for L-Dopa-induced abnormal involuntary movements (AIMs) and rotations following M1 microinfusions of +/-8-OH DPAT with or without coadministration of the 5-HT1AR antagonist WAY100635. Finally, +/-8-OH-DPAT was infused into M1 at peak dyskinesia to determine if 5 HT1AR stimulation attenuates established L-Dopa-induced AIMs and rotations. While no treatment effects were seen within the PFC, systemic +/-8-OH-DPAT suppressed L Dopa-induced c-fos within M1. Intra-M1 5-HT1AR stimulation diminished the onset of AIMs and this effect was reversed by WAY100635 indicating receptor specific effects. Finally, continuous infusion of +/-8-OH-DPAT into M1 at peak dyskinesia alleviated L-Dopa-induced AIMs. Collectively, these findings support an integral role for M1 in LID and its modulation by local 5-HT1AR. PMID- 21635908 TI - Low dose methamphetamine mediates neuroprotection through a PI3K-AKT pathway. AB - High doses of methamphetamine induce the excessive release of dopamine resulting in neurotoxicity. However, moderate activation of dopamine receptors can promote neuroprotection. Therefore, we used in vitro and in vivo models of stroke to test the hypothesis that low doses of methamphetamine could induce neuroprotection. We demonstrate that methamphetamine does induce a robust, dose-dependent, neuroprotective response in rat organotypic hippocampal slice cultures exposed to oxygen-glucose deprivation (OGD). A similar dose dependant neuroprotective effect was observed in rats that received an embolic middle cerebral artery occlusion (MCAO). Significant improvements in behavioral outcomes were observed in rats when methamphetamine administration delayed for up to 12 h after MCAO. Methamphetamine-mediated neuroprotection was significantly reduced in slice cultures by the addition of D1 and D2 dopamine receptor antagonist. Treatment of slice cultures with methamphetamine resulted in the dopamine-mediated activation of AKT in a PI3K dependant manner. A similar increase in phosphorylated AKT was observed in the striatum, cortex and hippocampus of methamphetamine treated rats following MCAO. Methamphetamine-mediated neuroprotection was lost in rats when PI3K activity was blocked by wortmannin. Finally, methamphetamine treatment decreased both cleaved caspase 3 levels in slice cultures following OGD and TUNEL staining within the striatum and cortex in rats following transient MCAO. These data indicate that methamphetamine can mediate neuroprotection through activation of a dopamine/PI3K/AKT-signaling pathway. PMID- 21635909 TI - Immune function and HPA axis activity in free-ranging rhesus macaques. AB - In mammals, the hypothalamic-pituitary-adrenal (HPA) axis and immune system play an important role in the maintenance of homeostasis. Dysregulation of either system resulting, for example, from psychosocial or reproductive stress increases susceptibility to disease and mortality risk, especially in aging individuals. In a study of free-ranging rhesus macaques, we examined how female age, reproductive state, social rank, and body condition influence (i) aspects of cytokine biology (plasma concentrations of interleukin-1 receptor antagonist (IL-1ra), IL-6 and IL 8), and (ii) HPA axis activity (plasma and fecal glucocorticoid levels). We also assessed individual differences in cytokine and hormone concentrations over time to determine their consistency and to investigate relations between these two indicators of physiological regulation and demand. Female monkeys showed marked increases in HPA axis activity during pregnancy and lactation, and increased circulating levels of IL-1ra with advancing age. Inter-individual differences in IL-1ra and IL-8 were consistent over successive years, suggesting that both are stable, trait-like characteristics. Furthermore, the concentrations of fecal glucocorticoid hormones in non-pregnant, non-lactating females were correlated with their plasma cortisol and IL-8 concentrations. Some individuals showed permanently elevated cytokine levels or HPA axis activity, or a combination of the two, suggesting chronic stress or disease. Our results enhance our understanding of within- and between-individual variation in cytokine levels and their relationship with glucocorticoid hormones in free-ranging primates. These findings can provide the basis for future research on stress and allostatic load in primates. PMID- 21635910 TI - Rats that binge eat fat-rich food do not show somatic signs or anxiety associated with opiate-like withdrawal: implications for nutrient-specific food addiction behaviors. AB - Previous studies suggest that binge eating sugar leads to behavioral and neurochemical changes similar to those seen with drug addiction, including signs of opiate-like withdrawal. Studies are emerging that show multiple neurochemical and behavioral indices of addiction when animals overeat a fat-rich diet. The goal of the present study was to utilize liquid and solid diets high in sugar and fat content to determine whether opiate-like withdrawal is seen after binge consumption of these diets in Sprague-Dawley rats. Control groups were given ad libitum access to the sweet-fat food or standard chow. All rats were then given a battery of tests to measure signs of opiate-like withdrawal, which included somatic signs of distress, elevated plus-maze anxiety, and locomotor hypoactivity. Neither naloxone-precipitated (3 mg/kg) nor deprivation-induced withdrawal was observed in rats that were maintained on a nutritionally complete pelleted sweet-fat diet or a sweet, high-fat diet supplemented with standard rodent chow. Naloxone-precipitated withdrawal was also not seen in rats fed a liquid sweet-fat food. Further, body weight reduction to 85%, which is known to potentiate the reinforcing effects of substances of abuse, did not affect naloxone-precipitated signs of opiate-like withdrawal. Thus, unlike previous findings reported regarding rats with binge access to a sucrose solution, rats that binge eat sweet-fat combinations do not show signs of opiate-like withdrawal under the conditions tested. These data support the idea that excessive consumption of different nutrients can induce behaviors associated with addiction in different ways, and that the behaviors that could characterize "food addiction" may be subtyped based on the nutritional composition of the food consumed. PMID- 21635911 TI - The influence of environmental bacteria in freshwater stingray wound-healing. AB - Invasion by bacteria can influence the course of healing of wounds acquired in aquatic environment. In this study, the bacteria present in Potamotrygon motoro stingray mucus and in the Alto Parana river water were identified, and their ability to induce tissue injury and resist antibiotics was determined. Biochemical identification analysis showed that 97% of all bacterial isolates were Gram negative, Aeromonas spp., Enterobacter cloacae and Citrobacter freundii being the species most prevalent. Gelatinase and caseinase were produced by Aeromonas hydrophila, Aeromonas sobria and Pseudomonas aeruginosa strains. Erythrocyte hemolysis assay showed that A. sobria, A. hydrophila and to a lesser extent, other Gram-negative bacteria produced hemolysin. It was also observed that molecules released in culture by these bacteria were toxic to human epithelial cells. Antibiogram results showed that 68% of all bacterial isolates were resistant to at least one type of antibiotic, mainly B-lactams. Finally, it was demonstrated that although P. motoro venom was toxic to epithelial cells it did not influence bacterial proliferation. In summary, the results obtained in this work indicate that during the accident, the mucus of P. motoro and the environmental water may transfer into the wound pathogenic multi-resistant bacteria with the potential to cause severe secondary infections. PMID- 21635912 TI - Anticancer activity of a low immunogenic protein toxin (BMP1) from Indian toad (Bufo melanostictus, Schneider) skin extract. AB - Earlier, a protein (BMP1, MW-79kDa) had been isolated from Indian toad (Bufo melanostictus) skin aqueous extract possessed anticancer activity against EAC bearing mice (Bhattacharjee et al., 2011). In the present study, the anti proliferative and apoptogenic activities of BMP1 have been evaluated in leukemic (U937 and K562) and hepatoma (HepG2) cells. BMP1 dose dependently inhibited U937 and K562 cell growth having IC50 values of 49 MUg/ml and 30 MUg/ml respectively. The anti-proliferative activity of BMP1 was observed in MTT assay, proliferating cell nuclear antigen (PCNA) expression and cell cycle arrest study. Flow cytometric data revealed that BMP1 arrested cell cycle in U937 and K562 cells at Sub-G1 and G1 phases. The BMP1-induced dose dependent expressions of CDKIs (p21(cip1) and p27(kip1)) and inhibition of CDK2 and PCNA expression in HepG2 cells support the inhibition of cell proliferation due to G1 arrest. BMP1-induced apoptosis analyzed by annexin-V binding study and the DNA fragmentation by comet assay were correlated with the sub-G1 arrest. The parallel induction of bax and p53 expression in HepG2 cells and the up-regulation of caspase 3 and caspase 9 due to BMP1 treatment indicated the involvement of p53-dependent intrinsic pathway of apoptosis. BMP1 was found to be low immunogenic in nature. PMID- 21635913 TI - Oral toxicity of extracts of the microcystin-containing cyanobacterium Planktothrix agardhii to the medaka fish (Oryzias latipes). AB - As previously demonstrated the medaka fish appears to offer a good model for studies of microcystins (MCs) effects. Since cyanobacterial toxins are released with other molecules in the aquatic environment when the producers are dying, in this study, we performed additional experiments in order to compare the described effects obtained with the pure toxin microcystin-LR (MC-LR), among the most toxic MCs, to those induced by complex extracts of an MCs-producer Planktothrix agardhii, strain PMC 75.02 and a natural bloom containing the MCs-producer P. agardhii. The toxicity of these extracts containing several variants of MC was determined in adult medaka treated by gavage. Extracts of an MCs-free strain of P. agardhii (PMC 87.02) were assayed for comparison. Extracts effects were analysed on two tissues, liver and intestine by means of photon and transmission electron microscopy. MC was localized in these tissues by immunocytochemistry. No effect was detectable with extracts of the MCs-free P. agardhii strain. The two MCs-P. agardhii extracts (strain and natural bloom) were able to induce harmful effects in the liver and intestine of the medaka fish in acute intoxication by gavage. In these target organs as shown by toxin immunolocalization, reactions leading to cell disjunction and lysis were observed apparently associated with an immune reaction implying MC containing macrophages. These effects are similar to those previously described with photonic microscopy in medaka treated with pure MC-LR with additional results obtained under the electron microscope. Since no significant effect was detected with the MCs-free (PMC 87.02) extract, we then conclude that MCs, even in complex association with other cyanobacterial components, should be responsible for the toxic effects observed in treated fish. PMID- 21635914 TI - Chronic cocaine self-administration attenuates the anxiogenic-like and stress potentiating effects of the benzodiazepine inverse agonist, FG 7142. AB - Stress is a well-known risk factor in relapse to drug abuse. Several forms of stress in animals have been used with varied degrees of success to elicit reinstatement of drug-seeking after chronic drug self-administration. Here, we tested the ability of the benzodiazepine (BZ) inverse agonist, FG 7142, to elicit anxiety-like behavior and potentiate stress responses in rats as measured by standard behavioral and hormonal indices and for its ability to affect reinstatement of cocaine-seeking in rats with a prior history of cocaine self administration. FG 7142 elicited anxiety-like behavior on the elevated plus maze (EPM) in cocaine-naive rats, and cocaine-naive rats injected with FG 7142 exhibited increased plasma corticosterone levels following EPM exposure. However, in animals with a history of cocaine self-administration, FG 7142 failed to affect elevated plus maze performance and did not affect plasma corticosterone response to the EPM. Furthermore, FG 7142 failed to reinstate cocaine-seeking, nor did it alter conditioned cue-induced reinstatement. These data indicate that the anxiety-related and stress potentiating qualities of BZ inverse agonism are attenuated in cocaine-experienced animals and do not lead to reinstatement of cocaine-seeking. PMID- 21635915 TI - Interaction studies of aristolochic acid I with human serum albumin and the binding site of aristolochic acid I in subdomain IIA. AB - Optical spectroscopy and molecular docking methods were used to examine the binding of aristolochic acid I (AAI) to human serum albumin (HSA) in this paper. By monitoring the intrinsic fluorescence of single Trp214 residue and performing displacement measurements, the specific binding of AAI in the vicinity of Sudlow's Site I of HSA has been clarified. An apparent distance of 2.53nm between the Trp214 and AAI was obtained via fluorescence resonance energy transfer (FRET) method. In addition, the changes in the secondary structure of HSA after its complexation with the ligand were studied with circular dichroism (CD) spectroscopy, which indicated that AAI does not has remarkable effect on the structure of the protein. Moreover, thermal denaturation experiments clearly indicated that the HSA-AAI complexes are conformationally more stable. Finally, the binding details between AAI and HSA were further confirmed by molecular docking studies, which revealed that AAI was bound at subdomain IIA through multiple interactions, such as hydrophobic effect, van der Waals forces and hydrogen bonding. PMID- 21635916 TI - Sodium alginate/poly(vinyl alcohol)/nano ZnO composite nanofibers for antibacterial wound dressings. AB - Sodium alginate (SA)/poly (vinyl alcohol) (PVA) fibrous mats were prepared by electrospinning technique. ZnO nanoparticles of size ~160nm was synthesized and characterized by UV spectroscopy, dynamic light scattering (DLS), XRD and infrared spectroscopy (IR). SA/PVA electrospinning was further carried out with ZnO with different concentrations (0.5, 1, 2 and 5%) to get SA/PVA/ZnO composite nanofibers. The prepared composite nanofibers were characterized using FT-IR, XRD, TGA and SEM studies. Cytotoxicity studies performed to examine the cytocompatibility of bare and composite SA/PVA fibers indicate that those with 0.5 and 1% ZnO concentrations are less toxic where as those with higher concentrations of ZnO is toxic in nature. Cell adhesion potential of this mats were further proved by studying with L929 cells for different time intervals. Antibacterial activity of SA/PVA/ZnO mats were examined with two different bacteria strains; Staphylococcus aureus and Escherichia coli, and found that SA/PVA/ZnO mats shows antibacterial activity due to the presence of ZnO. Our results suggest that this could be an ideal biomaterial for wound dressing applications once the optimal concentration of ZnO which will give least toxicity while providing maximum antibacterial activity is identified.f. PMID- 21635917 TI - Early stage aggregation of human serum albumin in the presence of metal ions. AB - The heat induced aggregation of human serum albumin (HSA) with and without an equimolar amount of Cu(II) and Zn(II) was investigated by using optical absorption, fluorescence, AFM and EPR spectroscopy. Turbidity experiments as a function of temperature indicate that the protein aggregation occurs after the melting of the protein. The kinetic of HSA aggregation, investigated between 60 and 70 degrees C by monitoring the optical density changes at 400nm on a 180min time window, shows an exponential growth with a rate that increases with the temperature. Fluorescence of the thioflavin T evidences a significant increase of the intensity at 480nm at increasing incubation time. These results combined with AFM experiments show that the protein aggregates are elongated oligomers with fibrillar-like features. The absence of a lag-phase suggests that the early stage aggregation of HSA follows a downhill pathway that does not require the formation of an organized nucleus. The presence of Cu(II) and Zn(II) ions does not affect the thermally induced aggregation process and the morphology of HSA aggregates. The result is compatible with the binding of the metal ions to the protein in the native state and with the high conformational stability of HSA. PMID- 21635918 TI - Noninvasive pulmonary function screening in spontaneously breathing rodents: an engineering systems perspective. AB - Noninvasive pulmonary function measurements made on rodents are commonly used for studies where quick, relatively easy end-points are required. These types of measurements are of particular advantage for studies where large numbers of animals are involved. Using tests that are simple to administer generally translates to more efficient and more accurate data collection. Noninvasive measurements result in less stress placed on the animal and allow repeated testing of the same animals at multiple time points. This review focuses on several noninvasive methods that have been developed for pulmonary function screening, which are analyzed from an engineering systems perspective. An analog model of the respiratory system of a conscious, freely respiring animal is presented in terms of an equivalent electrical circuit. This model is used as a basis to demonstrate the relationship between pulmonary parameters derived from circuit analysis. PMID- 21635919 TI - Congenital adrenal hyperplasia--pharmacologic interventions from the prenatal phase to adulthood. AB - Congenital adrenal hyperplasia (CAH) is one of the most common inherited autosomal recessive disorders, caused by deficiency of one of the enzymes involved in steroid synthesis. The clinical picture of the most prevalent form, i.e. 21-hydroxylase deficiency, is characterized by cortisol and mostly aldosterone deficiency and androgen excess (leading to congenital virilization in girls). Treatment consists of glucocorticoids, aimed at substitution of cortisol deficiency and, decrease of androgen excess. Usually supraphysiological doses of glucocorticoids are required to effectively suppress adrenal androgens. Furthermore, with the currently available glucocorticoid preparations, it is not possible to simulate a normal circadian rhythm in CAH patients. Therefore, it is a difficult task for (pediatric) endocrinologists to find the best balance between under- and overtreatment thereby avoiding important long term complications. In this review we will discuss the current pharmacologic treatment options. We give age dependent dose recommendations and describe the limitations of current treatment strategies. We discuss effects on fertility, bone density and cardiovascular risks. Recommendations about the use of glucocorticoids in case of fever or stress situations are given. The principles of treatment of non classic (mild) CAH are discussed in a separate section. Also prenatal therapy, to prevent congenital virilization of a female CAH newborn, is discussed. Furthermore, an overview of alternative pharmacological treatment options in the future is given. PMID- 21635920 TI - Performance of six commercial enzyme immunoassays and two alternative HIV-testing algorithms for the diagnosis of HIV-1 infection in Kisumu, Western Kenya. AB - Performances of serological parallel and serial testing algorithms were analyzed using a combination of three ELISA and three rapid tests for the confirmation of HIV infection. Each was assessed individually for their sensitivity and specificity on a blinded panel of 769 retrospective sera of known HIV status. Western blot was used as a confirmatory assay for discordant results. Subsequently, one parallel and one serial testing algorithm were assessed on a new panel of 912 HIV-positive and negative samples. Individual evaluation of the ELISAs and rapid tests indicated a sensitivity of 100% for all assays except Uni Gold with 99.7%. The specificities ranged from 99.1% to 99.4% for rapid assays and from 97.5% to 99.1% for ELISAs. A parallel and serial testing algorithms using Enzygnost and Vironostika, and Determine followed by Uni-Gold respectively, showed 100% sensitivity and specificity. The cost for testing 912 samples was US$4.74 and US$ 1.9 per sample in parallel and serial testing respectively. Parallel or serial testing algorithm yielded a sensitivity and specificity of 100%. This alternative algorithm is reliable and reduces the occurrence of both false negatives and positives. The serial testing algorithm was more cost effective for diagnosing HIV infections in this population. PMID- 21635921 TI - Validation of a recombinant integrin alphavbeta6/monoclonal antibody based antigen ELISA for the diagnosis of foot-and-mouth disease. AB - A sandwich ELISA using recombinant integrin alphavbeta6 as a capture ligand and serotype-specific monoclonal antibodies (Mabs) as detecting reagents has been compared with a polyclonal antibody based ELISA (using type-specific rabbit antibodies as capture and guinea pig antibodies as detectors), which is employed routinely at the FAO World Reference Laboratory for Foot-and-Mouth Disease (FMD), for the identification and serotyping of FMD virus (FMDV). The study used cell culture grown antigens (1351 FMDV positive) derived from suspected cases of vesicular disease collected from 86 countries between 1924 and 2011, those positive for the other vesicular diseases of swine vesicular disease (n = 25) and vesicular stomatitis (n = 45) and negative samples collected from uninfected cell cultures (n = 36). The diagnostic sensitivity of the assays was similar at 98.1% (polyclonal ELISA) compared to 97.9% (integrin/Mab ELISA) but the serotypic specificity of the latter was vastly superior (96%) to that of the former (61.5%). Reactions with the viruses of swine vesicular disease and vesicular stomatitis, which produce clinically indistinguishable syndromes in pigs and cattle, did not occur. The integrin/Mab ELISA recognized FMDV strains of wide antigenic and molecular diversity of all seven serotypes and although some FMDV isolates were not detected, the greater specificity of the assay, while retaining test sensitivity comparable to the conventional assay, warrants its consideration for adoption for routine diagnostic use. PMID- 21635922 TI - Species-specific identification of variola, monkeypox, cowpox, and vaccinia viruses by multiplex real-time PCR assay. AB - A method of one-stage rapid identification of variola (VARV), monkeypox (MPXV), cowpox (CPXV), and vaccinia (VACV) viruses, pathogenic for humans, utilizing multiplex real-time TaqMan PCR (MuRT-PCR) assay was developed. Four pairs of oligonucleotide primers and four hybridization probes with various fluorescent dyes and the corresponding fluorescence quenchers were concurrently used for MuRT PCR assay. The hybridization probe specific for the VARV sequence contained FAM/BHQ1 as a dye/quencher pair; MPXV-specific, TAMRA/BHQ2; CPXV-specific, JOE/BHQ1; VACV-specific, Cy5/BHQ3. The specificity and sensitivity of the developed method were assessed by analyzing DNA of 29 strains belonging to six orthopoxvirus species as well as the DNA samples isolated from archive clinical specimens of human smallpox cases and experimental specimens isolated from CPXV infected mice and MPXV-infected marmot. PMID- 21635923 TI - Detection, discrimination and absolute quantitation of Tomato spotted wilt virus isolates using real time RT-PCR with TaqMan((r))MGB probes. AB - A quantitative real-time reverse transcription-polymerase chain reaction (RT qPCR) procedure using a general primer set and three TaqMan((r))MGB probes was developed for general and genotype-specific detection and quantitation of the genomic M segment of Tomato spotted wilt virus (TSWV). Standard curves using RNA transcripts homologous to the three probes allowed reproducible quantitative assays with a wide dynamic range (10(3)-10(10) TSWV M segment RNA copies/ng of total RNA) and high sensitivity. This protocol was assayed with a battery of TSWV isolates, covering the range of the present known genetic variation, in single and/or mix infections in three plant hosts, as well as in the thrips vector Frankliniella occidentalis. This quantitative detection assay will be a valuable tool for molecular biology and epidemiology studies, diagnosis and disease control. PMID- 21635924 TI - Assessment of the efficacy of the neuraminidase inhibitor oseltamivir against 2009 pandemic H1N1 influenza virus in ferrets. AB - Pandemic 2009 influenza A (H1N1) virus (H1N1pdm) is different from contemporary seasonal human viruses in that it can cause infection deep in the lungs of critical care patients. Here we establish a mammalian animal model and assessed the efficacy of the neuraminidase (NA) inhibitor oseltamivir treatment against H1N1pdm virus infection. Oseltamivir (25 mg/kg/day twice daily for 5 days) was orally administered to groups of ferrets, starting either 2 or 24 h after inoculation with 10(6)PFU of A/California/04/2009 (H1N1) influenza virus. We determined that virus replication was restricted to 1 or 2 of 4 lung lobes in oseltamivir-treated animals, while virus was consistently isolated from 4 of 4 lung lobes in control animals (1.5-3.8log(10)PFU/g). Analysis of arterial blood oxygenation revealed less pronounced changes in partial oxygen and carbon dioxide pressure in oseltamivir-treated ferrets, and histologic examination confirmed reduced pneumonia. Treated animals had significantly decreased inflammatory responses in the upper respiratory tract (P < 0.05), less fever and weight loss, and less reduction of activity. Virus titers in the nasal washes of treated and control ferrets did not differ significantly. NA sequencing and fluorescence based phenotypic assays identified no oseltamivir-resistant variants. Overall, oseltamivir treatment decreases the signs of infection and reduced the spread of H1N1pdm influenza virus in the lungs of ferrets and therefore impeded the development of viral pneumonia. PMID- 21635925 TI - High serum fasting peptide YY (3-36) is associated with obesity-associated insulin resistance and type 2 diabetes. AB - We studied whether serum fasting levels of active form of peptide YY (PYY), PYY(3 36), are associated with obesity and related phenotypes. The study population consisted of 428 patients with coronary artery disease and diagnosed type 2 diabetes and 440 patients with coronary artery disease but without evidence of diabetes from the ARTEMIS study. The patients were recruited from the consecutive series of patients undergoing coronary angiography in the Oulu University Hospital. The patients without diabetes underwent a 2-hour oral glucose tolerance test. PYY(3-36) levels were analyzed by human PYY(3-36) specific radioimmunoassay. Result suggested that when PYY(3-36) tertiles were considered, high serum fasting PYY(3-36) concentration was associated with high body mass index, waist circumference, hemoglobin A1c, fasting blood glucose, leptin, triglyceride (p for all p <= 0.001), serum insulin (p=0.013) and with a low high density lipoprotein cholesterol (p=0.004) concentrations in the analyses adjusted for age, sex and study group. The link high PYY(3-36)-high insulin level was evident in subjects with normal glucose tolerance (p<0.05). The prevalence of diabetes was 72%, 46% and 30% in the highest, medium and lowest PYY(3-36) tertile (p<0.001). The PYY(3-36) concentrations (after adjustment for age, sex and body mass index) were higher in type 2 diabetics compared to subjects with impaired fasting glucose, impaired glucose tolerance and normal glucose tolerance (p<0.001 for trend). In conclusion, fasting PYY(3-36) concentrations in type 2 diabetic subjects are high. Although high PYY(3-36) is strongly linked to obesity and associated insulin resistance, the relation between PYY(3-36) and type 2 diabetes is independent of body fatness. PMID- 21635926 TI - Intracellular reactivation of transcription factors fused with protein transduction domain. AB - Induction of a desired cell type by defined transcription factors (TFs) using iPS technology can be used for cell replacement therapy. However, to overcome problems such as tumor formation, genomic insertional mutagenesis by viral transduction in the induction process needs to be avoided using alternative approaches. One approach could be the direct delivery of TF protein by a protein transduction system, whereby a protein transduction domain (PTD) is fused to facilitate the penetration of cell membrane. However, fusion proteins, including TFs, are reported to be biologically less active through the interference of PTD with proper protein folding. Here, we report a proof-of-concept study in which TF proteins fused with PTDs could be reactivated by removal of PTDs from cells. We demonstrated that Sox2 and Oct3/4 proteins fused with PTD were less active in mouse embryonic stem cells. Removal of PTD by a site-specific protease, derived from tobacco etch virus (TEV), substantially restored the functionality of these proteins, proved by enhanced rescue ability for differentiation induced by endogenous Sox2 and Oct3/4 repression. These results suggest that, by removing a PTD inside the cells, directly delivered TF proteins may exert substantially enhanced function than presently considered. PMID- 21635927 TI - Plant viruses: exploiting agricultural and natural ecosystems. PMID- 21635928 TI - "You Say it's Liking, I Say it's Wanting ...". On the difficulty of disentangling food reward in man. AB - According to the influential theory of Berridge (1996, 2009), food reward comprises two components: food 'liking' and 'wanting', with 'liking' referring to the pleasure derived from eating a given food and 'wanting' referring to appetitive motivation. Animal research shows that these two components have separate neural correlates. In examining reward driven eating in man, researchers have thus begun to develop interest in indicators of 'liking' and 'wanting'. But validating 'liking' and 'wanting' requires the dissociation of these components when theory dictates they should diverge. One such circumstance is neural sensitization as this leads to exaggerated 'wanting' without increased 'liking'. However, there are no data suggesting that such sensitization is the likely result of (over)eating. Without sensitization, one cannot determine whether task performance is indicative of true food 'liking' or 'wanting'. It is concluded that it is important to assess appetite and palatability in the study of reward driven eating, but determining whether these measurements reflect either food 'wanting' or food 'liking' is not. PMID- 21635929 TI - Therapeutic applications of ghrelin to cachexia utilizing its appetite stimulating effect. AB - Ghrelin, which is a natural ligand for the growth hormone (GH)-secretagogue receptor (GHS-R), stimulates food intake in both animals and humans. Ghrelin is the only circulating hormone known to stimulate appetite in humans. Ghrelin also stimulates GH secretion and inhibits the production of anorectic proinflammatory cytokines. As GH is an anabolic hormone, protein stores are spared at the expense of fat during conditions of caloric restriction. Thus, ghrelin exhibits anti cachectic actions via both GH-dependent and -independent mechanisms. Several studies are evaluating the efficacy of ghrelin in the treatment of cachexia caused by a variety of diseases, including congestive heart failure, chronic obstructive pulmonary disease, cancer, and end-stage renal disease. These studies will hopefully lead to the development of novel therapeutic applications for ghrelin in the future. This review summarizes the recent advances in this area of research. PMID- 21635930 TI - Dynamics of diapause hormone and prothoracicotropic hormone transcript expression at diapause termination in pupae of the corn earworm, Helicoverpa zea. AB - Both diapause hormone (DH) and ecdysone (E) are capable of terminating pupal diapause in members of the Helicoverpa/Heliothis complex. In this study we examine how the transcript encoding prothoracicotropic hormone (PTTH), the neuropeptide that stimulates the prothoracic gland to produce E, and the transcript encoding DH respond to developmental changes, as well as environmental and hormonal cues that can trigger the termination of diapause. In nondiapausing individuals PTTH and DH transcripts are abundant from pupation until adult eclosion, while in pupae that enter diapause PTTH transcripts are undetectable and abundant DH transcripts are present only briefly after pupation. Injection of E can break diapause at either 18 or 21 degrees C, but DH is effective in breaking diapause only at the higher temperature. Transfer of pupae to a diapause terminating temperature of 25 degrees C, injections of 1nmol DH or 75ng E at 21 degrees C, and injections of 500ng E at 18 degrees C, are all accompanied by a simultaneous elevation of mRNAs encoding both PTTH and DH, although the rate of PTTH mRNA increase is consistently more rapid than that of DH. Subthreshold doses of E or injections of distilled water elicit a temporary rise in PTTH and DH transcripts but do not lead to diapause termination. The results suggest that these two hormonal systems work together in the cascade of events leading to diapause termination, producing a sophisticated control system that is finely tuned and responsive to subtle temperature changes in the overwintering environment. PMID- 21635931 TI - The mTOR signaling pathway in the prefrontal cortex is compromised in major depressive disorder. AB - Recent studies demonstrate that rapid antidepressant response to ketamine is mediated by activation of the mammalian target of rapamycin (mTOR) signaling pathway, leading to increased synaptic proteins in the prefrontal cortex (PFC) of rats. Our postmortem studies indicate robust deficits in prominent postsynaptic proteins including N-methyl-d-aspartate (NMDA) receptor subunits (NR2A, NR2B), metabotropic glutamate receptor subtype 5 (mGluR5) and postsynaptic density protein 95kDa (PSD-95) in the PFC in major depressive disorder (MDD). We hypothesize that deficits in the mTOR-dependent translation initiation pathway contribute to the molecular pathology seen in the PFC of MDD subjects, and that a rapid reversal of these abnormalities may underlie antidepressant activity. The majority of known translational regulation occurs at the level of initiation. mTOR regulates translation initiation via its downstream components: p70-kDa ribosomal protein S6 kinase (p70S6K), and eukaryotic initiation factors 4E and 4B (eIF4E and eIF4B). In this study, we examined the expression of mTOR and its core downstream signaling targets: p70S6K, eIF4E, and eIF4B in the PFC of 12 depressed subjects and 12 psychiatrically healthy controls using Western blot. Levels of eIF4E phosphorylated at serine 209 (p-eIF4E-Ser209) and eIF4B phosphorylated at serine 504 (p-eIF4B-Ser504) were also examined. Adjacent cortical tissue samples from both cohorts of subjects were used in our previous postmortem analyses. There was a significant reduction in mTOR, p70S6K, eIF4B and p-eIF4B protein expression in MDD subjects relative to controls. No group differences were observed in eIF4E, p-eIF4E or actin levels. Our findings show deficits in mTOR dependent translation initiation in MDD particularly via the p70S6K/eIF4B pathway, and indicate a potential association between marked deficits in synaptic proteins and dysregulation of mTOR signaling in MDD. PMID- 21635932 TI - Biomarkers of exposure to triclocarban in urine and serum. AB - 3,4,4'-Trichlorocarbanilide (triclocarban, TCC) is widely used as an antimicrobial agent in a variety of consumer and personal care products. TCC is considered a potential endocrine disruptor, but its potential toxic effects in humans are still largely unknown. Because of its widespread uses, the potential for human exposure to TCC is high. In order to identify adequate exposure biomarkers of TCC, we investigated the metabolic profile of TCC in adult female Sprague Dawley rats after administering TCC once (500 mg/kg body weight) by oral gavage. Urine was collected 0-24 h before dosing, and 0-24 h and 24-48 h after dosing. Serum was collected at necropsy 48 h after dosing. We identified several metabolites of TCC in urine and serum by on-line solid phase extraction-high performance liquid chromatography-mass spectrometry. We unambiguously identified two major oxidative metabolites of TCC, 3'-hydroxy-TCC and 2'-hydroxy-TCC, by comparing their chromatographic behavior and mass spectral fragmentation patterns with those of authentic standards. By contrast, compared to these oxidative metabolites, we detected very low levels of TCC in the urine or serum. Taken together these data suggest that in rats, oxidation of TCC is a major metabolic pathway. We also measured TCC and its oxidative metabolites in 50 urine and 16 serum samples collected from adults in the United States. The results suggest differences in the metabolic profile of TCC in rats and in humans; oxidation appears to be a minor metabolic pathway in humans. Total (free plus conjugated) TCC could serve as a potential biomarker for human exposure to TCC. PMID- 21635933 TI - A structural analysis of G-quadruplex/ligand interactions. AB - This focused review article discusses in detail, all available high-resolution small molecule ligand/G-quadruplex structural data derived from crystallographic and NMR based techniques, in an attempt to understand key factors in ligand binding and to highlight the biological importance of these complexes. In contrast to duplex DNA, G-quadruplexes are four-stranded nucleic acid structures folded from guanine rich repeat sequences stabilized by the stacking of guanine G quartets and extensive Watson-Crick/Hoogsteen hydrogen bonding. Thermally stable, these topologies can play a role in telomere regulation and gene expression. The core structures of G-quadruplexes form stable scaffolds while the loops have been shown, by the addition of small molecule ligands, to be sufficiently adaptable to generate new and extended binding platforms for ligands to associate, either by extending G-quartet surfaces or by forming additional planar dinucleotide pairings. Many of these structurally characterised loop rearrangements were totally unexpected opening up new opportunities for the design of selective ligands. However these rearrangements do significantly complicate attempts to rationally design ligands against well defined but unbound topologies, as seen for the series of napthalene diimides complexes. Drawing together previous findings and with the introduction of two new crystallographic quadruplex/ligand structures we aim to expand the understanding of possible structural adaptations available to quadruplexes in the presence of ligands, thereby aiding in the design of new selective entities. PMID- 21635934 TI - Promiscuity, stability and cold adaptation of a newly isolated acylaminoacyl peptidase. AB - We report on the characterisation of a member of the acylaminoacyl peptidase family, the first isolated from bacteria. The enzyme was obtained from the psychrophilic bacterium Sporosarcina psychrophila and shows the typical features of cold adaptation (low T(m), optimal temperature of 40 degrees C, poor thermal stability). It was also tested for substrate specificity, effect of metals, temperature dependence and structure stability and revealed promiscuous catalytic activity on at least two chemically distinct substrates, with k(cat)/K(m) values for ester hydrolysis and acylamino acids cleavage of 1.7 * 10(4) s(-1) M(-1) and 6.2 * 10(3) s(-1) M(-1), respectively. Despite some properties cannot be explained with current models, results report on the relevance of structural and catalytic properties for the successful adaptation to cold temperatures. PMID- 21635935 TI - MHC class II upregulation and colocalization with Fas in experimental models of immune-mediated bone marrow failure. AB - OBJECTIVE: To test the hypothesis that gamma-interferon (IFN-gamma) promotes major histocompatibility complex (MHC) class II expression on bone marrow (BM) cell targets that facilitate T-cell-mediated BM destruction in immune-mediated BM failure. MATERIALS AND METHODS: Allogeneic lymph node (LN) cells were infused into MHC- or minor histocompatibility antigen-mismatched hosts to induce BM failure. MHC class II and Fas expression and cell apoptosis were analyzed by flow cytometry. MHC class II-Fas colocalization was detected by ImageStream Imaging Flow Cytometry and other cell-to-cell associations were visualized by confocal microscopy. T-cell-mediated BM cell apoptosis and effects of IFN-gamma on MHC class II-Fas colocalization on normal BM cells were studied using cell culture in vitro followed by conventional and imaging flow cytometry. RESULTS: BM failure animals had significantly upregulated MHC class II expression on CD4(-)CD8( )CD11b(-)CD45R(-) residual BM cells and significantly increased MHC class II-Fas colocalization on BM CD150(+) and CD34(+) hematopoietic cells. MHC class II(+)Fas(+) BM cells were closely associated with CD4(+) T cells in the BM of affected animals, and they were significantly more responsive to T-cell-mediated cell apoptosis relative to MHC class II(-)Fas(-) BM cells. Infusion of IFN-gamma deficient LN cells into minor histocompatibility antigen-mismatched recipients resulted in no MHC class II-Fas upregulation and no clinically overt BM failure. Treatment with recombinant IFN-gamma significantly increased both MHC class II Fas coexpression and colocalization on normal BM cells. CONCLUSIONS: Elevation of the inflammatory cytokine IFN-gamma-stimulated MHC class II expression and MHC class II-Fas colocalization, which may facilitate T-cell-mediated cell destruction. PMID- 21635936 TI - The neural organization of perception in chess experts. AB - The human visual system responds to expertise, and it has been suggested that regions that process faces also process other objects of expertise including chess boards by experts. We tested whether chess and face processing overlap in brain activity using fMRI. Chess experts and novices exhibited face selective areas, but these regions showed no selectivity to chess configurations relative to other stimuli. We next compared neural responses to chess and to scrambled chess displays to isolate areas relevant to expertise. Areas within the posterior cingulate, orbitofrontal cortex, and right temporal cortex were active in this comparison in experts over novices. We also compared chess and face responses within the posterior cingulate and found this area responsive to chess only in experts. These findings indicate that the configurations in chess are not strongly processed by face-selective regions that are selective for faces in individuals who have expertise in both domains. Further, the area most consistently involved in chess did not show overlap with faces. Overall, these results suggest that expert visual processing may be similar at the level of recognition, but need not show the same neural correlates. PMID- 21635937 TI - The significance of chloride in the inhibitory action of disodium cromoglycate on immunologically-stimulated rat peritoneal mast cells. AB - BACKGROUND: The microelectrode array (MEA) was used to investigate the pharmacological relevance of chloride (Cl-) ions in antigen-dependent mast cell activation and the inhibitory effect of disodium cromoglycate (DSCG) on mast cell activation. METHODS: The movements of ions across the cellular membrane and the potential relationship between Cl- channels and DSCG during immunological activation were investigated using the MEA. The results were then subsequently compared with the amount of histamine released from anti-IgE activated peritoneal mast cells. RESULTS: The inclusion of charybdotoxin (ChTX) in Cl--free buffer showed that the measured field potentials during antigen-stimulated peritoneal mast cell were a combination of Cl- influx and K+ efflux. The delayed onset time of Cl- influx indicated the presence of a delayed outwardly-rectifying Cl- current in the antigen-stimulated peritoneal mast cells. The use of 5-nitro-2-(3 phenylpropylamino) benzoic acid demonstrated that the activated mast cell membrane potential can be stabilised, thereby reducing the amount of histamine released from the anti-IgE activated mast cells. The correlation between the results of the histamine release assay and the electrophysiological measurements demonstrated the importance of Cl- to anti-IgE dependent mast cell activation. The inhibitory effect of DSCG on anti-IgE activated cells, however, did not correlate with the presumed influx of Cl-. CONCLUSIONS: The MEA data suggest that Cl- influx is crucial to IgE-dependent mast cell degranulation. GENERAL SIGNIFICANCE: While the MEA cannot yield information about single channel properties, it is convenient to use and can provide information on the global changes in electrophysiological responses of non-excitable cells. PMID- 21635938 TI - Increased sensitivity to the effects of chronic social defeat stress in an innately anxious mouse strain. AB - Stress and genetic predisposition are two of the major risk factors for a variety of psychiatric illnesses. Inbred mouse strains are considered useful tools in dissecting the genetic basis of complex disorders. Indeed, mice of the C57BL/6 and BALB/c strains, differing markedly in anxiety behaviours, are among the most widely used in psychopharmacological research. However, there is a paucity of studies investigating the impact of social stress in these two strains. Moreover, whether these two mouse strains exhibit different sensitivities to chronic social defeat stress remains poorly studied. Thus in this study we compared the impact of repeated (10 days) social defeat stress on a variety of behavioural and endocrine parameters including social interaction, locomotor activity, plasma corticosterone, body weight and stress-related physiological parameters in both mouse strains. Given that the duration of stress exposure may differentially affect such responses we also compared stressors of short (Social Defeat-Short; SD-S) and of long (Social Defeat-Long; SD-L) duration. Our results show that although mice from both strains were defeated in both social defeat paradigms, only BALB/c mice displayed social interaction impairments following SD-S, whereas both strains were behaviourally sensitive to SD-L. Moreover, both strains also differed in some of the physiological alterations induced by social defeat stress. Specifically, SD-S did not induce any change in corticosterone levels in either of the two strains, whereas SD-L was able to induce significant changes in C57BL/6 mice only. SD-S induced differential effects on bodyweight gain in both strains, increasing it in C57BL/6 and decreasing it in BALB/c mice, whereas SD-L had no effect. On the other hand, exposure to SD-S resulted in cardiac hypertrophy in C57BL/6 mice and SD-L induced spleen hypertrophy and thymus atrophy in BALB/c mice in addition to decreasing faecal output. Overall, the innately anxious BALB/c mice were more sensitive to social stress than C57BL/6, with differential behavioural and physiological alterations emerging as a function of stress severity. These data suggest different coping strategies to social interaction stress between the two mouse strains. The genetic basis of this stress-resilience/susceptibility warrants further investigation. PMID- 21635939 TI - Participation of the GABAergic system on the glutamate release of frontal cortex synaptosomes from Wistar rats with experimental autoimmune encephalomyelitis. AB - We previously found that the glutamate release was decreased in synaptosomes from rat cerebral cortex during the development of experimental autoimmune encephalomyelitis (EAE), the animal model of multiple sclerosis. Various other reports have shown a deficit in the expression of proteins associated with GABAergic neurotransmission in the neocortex of patients with multiple sclerosis and it was also demonstrated that the activation of GABAA receptors leads to an inhibition of glutamate release. Now, in order to evaluate the events that may affect the neuronal function in EAE synaptosomes, we analyzed the participation of the GABAergic system in glutamate release and in the flunitrazepam-sensitive GABAA receptor density. This revealed alterations in the GABAergic system of the frontal cortex synaptosomes from EAE animals. GABA induced a decrease in the 4 aminopyridine-evoked glutamate release in control synaptosomes which was abolished by picrotoxin, a GABAA receptor antagonist. In contrast, synaptosomes from EAE rats showed a loss in the inhibition of glutamate release mediated by GABA. Furthermore, the flunitrazepam-sensitive GABAA receptor density was decreased during the acute stage of the disease in synaptosomes from EAE rats. We also observed a loss of inhibition in the Ca2+-dependent phosphorylation of synapsin I mediated by GABA in nerve terminals from EAE animals, which could explain the loss of GABAergic regulation on evoked glutamate release. The changes observed in the GABAA receptor density as well as the loss of GABAergic inhibition of glutamate release were partially reverted in cortical synaptosomes from recovered EAE animals. These results suggest that the decrease in the flunitrazepam-sensitive GABAA receptor density may explain the observed failure of GABAergic regulation in the glutamate release of synaptosomes from EAE rats, which might contribute to the appearance of clinical symptoms and disease progression. PMID- 21635940 TI - A postmenopause-like model of ovariectomized Wistar rats to identify active principles of Erythrina lysistemon (Fabaceae). AB - To determine whether the two major compounds of Erythrina lysistemon are active principles accounting for Erythrina estrogenic effects, we used a postmenopause like model of ovariectomized Wistar rats to evaluate their effects on some menopausal problems. Ovariectomized rats were orally treated either with compound 1 or compound 2 at 1 and 10 mg/kg BW for 28 days. Estradiol valerate served as the reference substance. As results, compounds 1 and 2 displayed estrogen-like effects on the uterus and the vagina, and reduced atherogenic risks by decreasing the two assessed atherogenic parameters, the total cholesterol/HDL-cholesterol ratio and the atherogenic index of plasma. PMID- 21635941 TI - Irbic acid, a dicaffeoylquinic acid derivative from Centella asiatica cell cultures. AB - 3,5-O-dicaffeoyl-4-O-malonilquinic acid (1) (irbic acid) has been isolated for the first time from cell cultures of Centella asiatica and till now it has never been reported to be present in the intact plant. Evidence of its structure was obtained by spectroscopic analyses (MS/NMR). Besides 1, cell cultures produce also the known 3,5-O-dicaffeoylquinic acid, chlorogenic acid, and the triferulic acid 2 (4-O-8'/4'-O-8"-didehydrotriferulic acid). Biological activities were evaluated for compound 1, which showed to have a strong radical scavenging capacity, together with a high inhibitory activity on collagenase. This suggests a possible utilization of this substance as a topical agent to reduce the skin ageing process. PMID- 21635942 TI - TMPIT1 from wild emmer wheat: first characterisation of a stress-inducible integral membrane protein. AB - In this study a gene for a drought stress-inducible putative membrane protein was cloned and characterised from root tissue of wild emmer wheat. Sequence analysis indicated that the protein is a member of the widespread but hitherto uncharacterised TMPIT (transmembrane protein inducible by TNF-alpha) family, so it was labelled TdicTMPIT1. Real-time RT-PCR showed that the TdicTMPIT1 gene is upregulated on drought stress in drought-tolerant wild emmer wheat, but not in a drought-sensitive accession or in cultivated durum wheat. The TdicTMPIT1 product was predicted to be a membrane protein with four transmembrane helices. The protein was expressed and analysed in Escherichia coli and Saccharomyces cerevisiae. Cellular localisation of the protein in the cell was also investigated using an eGFP-tagged form of the protein in S. cerevisiae. Results obtained by confocal laser microscopy indicated that the TdicTMPIT1 tagged with GFP was localised in a membraneous compartment. It is concluded that TdicTMPIT1 is a membrane protein associated with the drought stress response in wild emmer wheat, and so it may be useful for the improvement of modern wheat genotypes. Members of this protein family in other organisms are proposed also to be involved in stress responses. PMID- 21635943 TI - Discrete element method (DEM) simulations of stratified sampling during solid dosage form manufacturing. AB - Discrete element model (DEM) simulations of the discharge of powders from hoppers under gravity were analyzed to provide estimates of dosage form content uniformity during the manufacture of solid dosage forms (tablets and capsules). For a system that exhibits moderate segregation the effects of sample size, number, and location within the batch were determined. The various sampling approaches were compared to current best-practices for sampling described in the Product Quality Research Institute (PQRI) Blend Uniformity Working Group (BUWG) guidelines. Sampling uniformly across the discharge process gave the most accurate results with respect to identifying segregation trends. Sigmoidal sampling (as recommended in the PQRI BUWG guidelines) tended to overestimate potential segregation issues, whereas truncated sampling (common in industrial practice) tended to underestimate them. The size of the sample had a major effect on the absolute potency RSD. The number of sampling locations (10 vs. 20) had very little effect on the trends in the data, and the number of samples analyzed at each location (1 vs. 3 vs. 7) had only a small effect for the sampling conditions examined. The results of this work provide greater understanding of the effect of different sampling approaches on the measured content uniformity of real dosage forms, and can help to guide the choice of appropriate sampling protocols. PMID- 21635944 TI - Oxidative mechanisms contribute to nanosize silican dioxide-induced developmental neurotoxicity in PC12 cells. AB - Neurotoxicity was investigated in nano-SiO2-treated cultured PC12 cells, an in vitro neuronal cell model, in order to define a relatively safe dose range for its application. The following were observed in the present study: (1) A dose dependent increase in the level of reactive oxygen species (ROS) with a corresponding decrease in the level of glutathione (R2=0.965) suggesting 20- and 50-nm SiO2-induced free radical generation and glutathione depletion. (2) A dose- and time-dependent decrease in cell viability that was associated with elevation of ROS level, especially after 24-h nano-SiO2 exposure (R2=0.965), suggesting the role of oxidative stress on nano-SiO2 induced cell death. (3) An increase in the level of thiobarbituric-acid reactive species that correlated reversely with cell viability of the PC12 cells treated with nano-SiO2 (R2=0.945) suggesting nano SiO2-induced membrane damage caused by lipid peroxidation. (4) A dose-dependent increase in sub-G1 population in SiO2-exposed cells along with cell shrinkage and nuclear condensation from morphological examination suggesting nano-SiO2-induced cell apoptosis. Furthermore, nano-SiO2 exposure diminished the ability of neurite extension in response to nerve growth factor in treated PC12 cells. In summary, SiO2 nanoparticle exposure resulted in dose-dependent neurotoxicity in cultured PC12 cells that was probably associated with oxidative stress and induced apoptosis. PMID- 21635945 TI - Lactate is an ideal non-invasive marker for evaluating temporal alterations in cell stress and toxicity in repeat dose testing regimes. AB - Technological developments are driving in vitro methods towards integrated "omic" strategies. However, there is still an over reliance on classical viability assays for dose range finding. Such assays are not readily suited to the investigation of subtle alterations in cell function and most require termination of the experiment, which makes it difficult to monitor temporal alterations in repeat-dose long term exposure experiments. To this end, we investigated the use of lactate production as a marker of cell stress in long term repeat dose experiments. We conducted daily exposures to eight compounds at five concentrations for 14 days on human renal proximal tubular cells (RPTEC/TERT1), human hepatoma cells (HepaRG) and mouse fibroblasts (BALB-3T3) cells. Compounds were chosen from a training set used in the 7th EU Framework project Predict-IV and consisted of amiodarone, diclofenac, troglitazone, cadmium chloride, cephaloridine, cidofovir, cyclosporine A and buflomedil. At days 1, 3, 7 and 14, lactate was measured in the supernatant medium. At day 14, cells were assayed for resazurin reduction capability and subsequently lysed in methanol for ATP determination. Compound-induced loss of viability was comparable across all cell lines. For all cell types, when cell viability was compromised at day 14, lactate production was induced during the treatment period. In some situations, lactate also fell below control values, indicating cell death. Thus, temporal alterations in supernatant lactate provides information on the time and concentration of stress induction and the time and concentration where cell death becomes the dominant factor. Supernatant lactate production is a simple, cheap and non invasive parameter. Since many molecular pathways converge on the glycolytic pathway, enhanced lactate production may be considered as a global marker of sub lethal injury and thus an ideal marker for investigating temporal alterations in long term repeat dose testing in vitro regimes. PMID- 21635946 TI - Lipopolysaccharide delays demyelination and promotes oligodendrocyte precursor proliferation in the central nervous system. AB - Systemic infection can influence the course in many diseases of the central nervous system (CNS) such as multiple sclerosis (MS), yet the relationship between infection outside the CNS and potential damage and/or protection within the CNS is still not understood. Activation of microglia is a characteristic feature of most CNS autoimmune disorders, including MS, and both protective and degenerative functions of microglia have been proposed. Hence, we analyzed the effects of a systemic inflammatory reaction induced by peripheral treatment with lipopolysaccharide (LPS) on microglial reaction and cuprizone induced de- and remyelination. We found that LPS administration delayed demyelination, which was linked with inhibition of microglial proliferation and reduced numbers of activated microglia. The phenotype of microglia changed as an increase of Toll like receptor 4 was found. During remyelination, LPS treatment delayed the onset of myelin protein re-expression, but later there was a beneficial effect via an increase of proliferating oligodendrocyte precursor cells (OPC) and mature oligodendrocytes. Moreover, the expression of ciliary neurotrophic factor was increased in response to LPS, a growth factor known to mediate OPC proliferation. Additional experiments showed that the time window to induce LPS effects was limited and associated with the presence of microglia. In conclusion, LPS delayed demyelination and caused beneficial effects on remyelination via increasing the proliferation of OPC. These differences seem to be an effect of LPS induced microglial modulation and indicate that exposure to certain infectious agents within a given time window may be beneficial in promoting tissue repair. PMID- 21635947 TI - Dissociation between learning and memory impairment and other sickness behaviours during simulated Mycoplasma infection in rats. AB - To investigate potential consequences for learning and memory, we have simulated the effects of Mycoplasma infection, in rats, by administering fibroblast stimulating lipopepide-1 (FSL-1), a pyrogenic moiety of Mycoplasma salivarium. We measured the effects on body temperature, cage activity, food intake, and on spatial learning and memory in a Morris Water Maze. Male Sprague-Dawley rats had radio transponders implanted to measure abdominal temperature and cage activity. After recovery, rats were assigned randomly to receive intraperitoneal (I.P.) injections of FSL-1 (500 or 1000 MUg kg(-1) in 1 ml kg(-1) phosphate-buffered saline; PBS) or vehicle (PBS, 1 ml kg(-1)). Body mass and food intake were measured daily. Training in the Maze commenced 18 h after injections and continued daily for four days. Spatial memory was assessed on the fifth day. In other rats, we measured concentrations of brain pro-inflammatory cytokines, interleukin (IL)-1beta and IL-6, at 3 and 18 h after injections. FSL-1 administration induced a dose-dependent fever (~1 degrees C) for two days, lethargy (~78%) for four days, anorexia (~65%) for three days and body mass stunting (~6%) for at least four days. Eighteen hours after FSL-1 administration, when concentrations of IL-1beta, but not that of IL-6, were elevated in both the hypothalamus and the hippocampus, and when rats were febrile, lethargic and anorexic, learning in the Maze was unaffected. There also was no memory impairment. Our results support emerging evidence that impaired learning and memory is not inevitable during simulated infection. PMID- 21635948 TI - The serotonergic system in fish. AB - Neurons using serotonin (5-HT) as neurotransmitter and/or modulator have been identified in the central nervous system in representatives from all vertebrate clades, including jawless, cartilaginous and ray-finned fishes. The aim of this review is to summarize our current knowledge about the anatomical organization of the central serotonergic system in fishes. Furthermore, selected key functions of 5-HT will be described. The main focus will be the adult brain of teleosts, in particular zebrafish, which is increasingly used as a model organism. It is used to answer not only genetic and developmental biology questions, but also issues concerning physiology, behavior and the underlying neuronal networks. The many evolutionary conserved features of zebrafish combined with the ever increasing number of genetic tools and its practical advantages promise great possibilities to increase our understanding of the serotonergic system. Further, comparative studies including several vertebrate species will provide us with interesting insights into the evolution of this important neurotransmitter system. PMID- 21635949 TI - Immunosuppressive mechanisms of embryonic stem cells and mesenchymal stem cells in alloimmune response. AB - Although both embryonic stem cells (ESCs) and mesenchymal stem cells (MSCs) are known to have immunosuppressive effects, the mechanisms of immunosuppression are still controversial. Both types of stem cells suppressed not only the proliferation but also survival of CD4(+) T cells in vitro. They suppressed secretion of various cytokines (IL-2, IL-12, IFN-gamma, TNF-alpha, IL-4, IL-5, IL 1beta, and IL-10), whereas there was no change in the levels of TGF-beta or IDO. Classic and modified transwell experiments demonstrated that immunosuppressive activities were mainly mediated by cell-to-cell contact. Granzyme B in the ESCs played a significant role in their immunosuppression, whereas PDL-1, Fas ligand, CD30 or perforin was not involved in the contact-dependent immunosuppression. However, none of the above molecules played a significant role in the immunosuppression by the MSCs. Interestingly, both stem cells increased the proportion of Foxp3(+) regulatory T cells. Our results showed that both ESCs and MSCs suppressed the survival as well as the proliferation of T cells by mainly contact-dependent mechanisms and increased the proportion of regulatory T cells. Granzyme B was involved in immunosuppression by the ESCs in a perforin independent manner. PMID- 21635950 TI - Distribution of intrahepatic T, NK and CD3(+)CD56(+)NKT cells alters after liver transplantation: Shift from innate to adaptive immunity? AB - BACKGROUND: The liver is an immunological organ containing a large number of T, NK and NKT cells, but little is known about intrahepatic immunity after LTx. Here, we investigated whether the distribution of T, NK and CD3(+)CD56(+)NKT cells is altered in transplanted livers under different circumstances. METHODS: Core biopsies of transplanted livers were stained with antibodies against CD3 and CD56. Several cell populations including T (CD3(+)CD56(-)), NK (CD3(-)CD56(+)) and NKT cells (CD3(+)CD56(+)) were studied by fluorescence microscopy. Cell numbers were analyzed in relation to the time interval after LTx, immunosuppressive therapy and stage of acute graft rejection (measured with the rejection activity index: RAI) compared to tumor free liver tissue from patients after liver resection due to metastatic disease as control. RESULTS: Recruitment of CD3(+)CD56(+)NKT cells revealed a significant decrease during high RAI scores in comparison to low and middle RAI scores (RAI 7-9: 0.03+/-0.01/HPF vs. RAI 4-6: 0.1+/-0.005/HPF). CD3(+)CD56(+)NKT cells were also lower during immunosuppressive therapy with tacrolimus (0.03+/-0.01/HPF) than with cyclosporine (0.1+/ 0.003/HPF), cyclosporine/MMF (0.1+/-0.003/HPF) or sirolimus (0.1+/-0.01/HPF) treatment. Intrahepatic T cell numbers increased significantly 50days after LTx compared to control liver tissue (4.5+/-0.2/HPF vs. 1.9+/-0.1/HPF). In contrast, NK cells (0.3+/-0.004/HPF) were significantly fewer in all biopsies after LTx compared to the control (0.7+/-0.04/HPF). CONCLUSIONS: These data indicate significant alterations in the hepatic recruitment of T, NK and CD3(+)CD56(+)NKT cells after LTx. The increase in T cells and the decrease in NK and CD3(+)CD56(+)NKT cells suggest a shift from innate to adaptive hepatic immunity in the liver graft. PMID- 21635951 TI - Nitric oxide, a janus-faced therapeutic target for diabetic microangiopathy Friend or foe? AB - Accelerated atherosclerosis and microvascular complications are the leading causes of coronary heart disease, end-stage renal failure, acquired blindness and a variety of neuropathy, which could account for disabilities and high mortality rates in patients with diabetes. As the prevalence of diabetes has risen to epidemic proportions worldwide, diabetic vascular complications have now become one of the most challenging health problems. Nitric oxide (NO) is a pleiotropic molecule critical to a number of physiological and pathological processes in humans. NO not only inhibits the inflammatory-proliferative reactions in vascular wall cells, but also exerts anti-thrombogenic and endothelial cell protective properties, all of which could potentially be exploited as a therapeutic option for the treatment of vascular complications in diabetes. However, high amounts of NO produced by inducible NO synthase (iNOS) and/or peroxynitrite (ONOO(-)), a reactive intermediate of NO with superoxide anion are involved in pro inflammatory reactions and tissue damage as well. This implies that NO is a janus faced molecule and acts as a double-edged sword in vascular complications in diabetes. Further, NO is synthesized from l-arginine via the action of NO synthase (NOS), while NOS is blocked by endogenous l-arginine analogues such as asymmetric dimethylarginine (ADMA), a naturally occurring amino acid which is found in the plasma and various tissues. These findings suggest that amounts of NO locally produced, oxidative stress conditions and level of ADMA could determine the beneficial and detrimental effects of NO on vascular complications in diabetes. In this paper, we review the janus-faced aspects of NO in diabetic microangiopathy. PMID- 21635952 TI - Kruppel-like transcription factors in the nervous system: novel players in neurite outgrowth and axon regeneration. AB - The Kruppel-like family of transcription factors (KLFs) have been widely studied in proliferating cells, though very little is known about their role in post mitotic cells, such as neurons. We have recently found that the KLFs play a role in regulating intrinsic axon growth ability in retinal ganglion cells (RGCs), a type of central nervous system (CNS) neuron. Previous KLF studies in other cell types suggest that there may be cell-type specific KLF expression patterns, and that their relative expression allows them to compete for binding sites, or to act redundantly to compensate for another's function. With at least 15 of 17 KLF family members expressed in neurons, it will be important for us to determine how this complex family functions to regulate the intricate gene programs of axon growth and regeneration. By further characterizing the mechanisms of the KLF family in the nervous system, we may better understand how they regulate neurite growth and axon regeneration. PMID- 21635953 TI - Evidence for altered hippocampal function in a mouse model of the human 22q11.2 microdeletion. AB - 22q11.2 chromosomal deletions are recurrent copy number mutations that increase the risk of schizophrenia around thirty-fold. Deletion of the orthologous chromosomal region in mice offers an opportunity to characterize changes to neuronal structure and function that may account for the development of this disease. The hippocampus has been implicated in schizophrenia pathogenesis, is reduced in volume in 22q11.2 deletion carriers and displays altered neuronal structure in a mouse model of the mutation (Df(16)A(+/-) mice). Here we investigate hippocampal CA1 physiology, hippocampal-dependent spatial memory and novelty-induced hippocampal activation in Df(16)A(+/-) mice. We found normal spatial reference memory (as assayed by the Morris water maze test) as well as modest but potentially important deficits in physiology. In particular, a reduction in the level of inhibition of CA1 pyramidal neurons was observed, implying a decrease in interneuron activity. Additionally, deficits in LTP were observed using certain induction protocols. Induction of c-Fos expression by exploration of a novel environment suggested a relative sparing of CA1 and dentate gyrus function but showed a robust decrease in the number of activated CA3 pyramidal neurons in Df(16)A(+/-) mice. Overall, experiments performed in this 22q11.2 deletion model demonstrated deficits of various degrees across different regions of the hippocampus, which together may contribute to the increased risk of developing schizophrenia. PMID- 21635954 TI - Structure and stability of the lamin A tail domain and HGPS mutant. AB - Hutchinson-Gilford progeria syndrome (HGPS) is a premature aging syndrome caused by the expression and accumulation of a mutant form of lamin A, Delta50 lamin A. As a component of the cell's nucleoskeleton, lamin A plays an important role in the mechanical stabilization of the nuclear envelope and in other nuclear functions. It is largely unknown how the characteristic 50 amino acid deletion affects the conformation of the mostly intrinsically disordered tail domain of lamin A. Here we perform replica exchange molecular dynamics simulations of the tail domain and determine an ensemble of semi-stable structures. Based on these structures we show that the ZMPSTE 24 cleavage site on the precursor form of the lamin A tail domain orients itself in such a way as to facilitate cleavage during the maturation process. We confirm our simulated structures by comparing the thermodynamic properties of the ensemble structures to in vitro stability measurements. Using this combination of experimental and computational techniques, we compare the size, heterogeneity of size, thermodynamic stability of the Ig-fold, as well as the mechanisms of force-induced denaturation. Our data shows that the Delta50 lamin A tail domain is more compact and displays less heterogeneity than the mature lamin A tail domain. Altogether these results suggest that the altered structure and stability of the tail domain can explain changed protein-protein and protein-DNA interactions and may represent an etiology of the disease. Also, this study provides the first molecular structure(s) of the lamin A tail domain, which is confirmed by thermodynamic tests in experiment. PMID- 21635955 TI - Population-wide principal component-based quantification of blood-brain-barrier dynamics in multiple sclerosis. AB - The processes by which new white matter lesions in multiple sclerosis (MS) develop are only partially understood. Much of this understanding has come through magnetic resonance imaging (MRI) of the human brain. One of the hallmarks of new lesion development in MS is enhancement on T(1)-weighted MRI scans following the intravenous administration of a gadolinium-based contrast agent that shortens the longitudinal relaxation time of the tissue. Visible enhancement on the MRI results from the opening of the blood-brain barrier and reveals areas of active inflammation. The incidence and number of existing enhancing lesions are common outcome measures used in MS treatment clinical trials. Dynamic contrast-enhanced MRI (DCE-MRI) can estimate the rate at which contrast agents pass from the plasma to MS lesions. In this paper, we develop a principal component-based framework for the analysis of these data that provides biologically meaningful quantification of blood-brain-barrier opening in new MS lesions. To accomplish this, we use functional principal components analysis to study directions of variation in the voxel-level time series of intensities both within and across subjects. The analysis reveals and allows quantification of typical spatiotemporal enhancement patterns in acute MS lesions, providing measures of magnitude, rate, shape (ring-like vs. nodular), and dynamics (centrifugal vs. centripetal). Across 10 subjects with relapsing-remitting and primary progressive MS, we found subjects to have between 0 and 12 gadolinium enhancing lesions, the majority of which enhanced centripetally. We quantified the spatiotemporal behavior within each of these lesions using novel measures. Further application of these techniques will determine the extent to which these lesion measures can predict or track response to therapy or long-term prognosis in this disorder. PMID- 21635956 TI - Heart rate correction models to detect QT interval prolongation in novel pharmaceutical development. AB - INTRODUCTION: The QT interval of the electrocardiogram (ECG) reflects the duration of ventricular depolarization and repolarization. A drug-induced prolongation of ventricular repolarization, and thereby QT prolongation, is recognized to be a marker for an enhanced risk for ventricular arrhythmia. The assessment of a drug's effect on the QT interval has therefore become routine within pharmaceutical research and development. However, the heart rate has a major influence on the QT interval; the QT interval shortens as heart rate increases such that one needs to account for such heart rate-dependent changes when evaluating possible drug-induced effects on the QT interval. The relationship between the QT interval and heart rate can be modeled mathematically and using this function a so-called "corrected" QT interval (QTc) can be generated to assess drug-induced effects independent from heart rate-dependent effects. In the past few years, a large number of mathematical relationship have been described that supposedly best describe the heart rate-QT relationship. In this paper we describe a novel approach for selecting the optimal mathematical function for this purpose for a given individual. METHODS: Mongrel, purpose-bred dogs (16, males and females) were instrumented with radiotelemetry transmitters (ITS) for measurement of aortic pressure (AP), left ventricular pressure (LVP), the lead II ECG and body temperature. ECGs were recorded continuously without drug treatment and include a range of HRs due to spontaneous, physiological changes over the 24h of data acquisition. Various mathematical models (>20) were then used to evaluate the HR-QT relationship and these were compared statistically to objectively select the model best fitting the data set of each individual animal. RESULTS: In this study a dynamic analysis algorithm was developed to find the optimal descriptor of the HR-QT relationship for a given individual animal under control conditions. The use of this optimal relationship provides the best possible approach for detecting drug-induced effects on the QT interval for compounds that also affect the heart rate. DISCUSSION: Several numerical methods to optimize the correction functions and statistical procedures to perform significance tests were discussed and implemented in a QT/RR relationship analysis system, named QTana. Given a sample data set, QTana searches the best correction model(s) from the integrated 11 QT/RR relationship modeling functions. PMID- 21635957 TI - Trends in safety pharmacology: posters presented at the annual meetings of the Safety Pharmacology Society 2001-2010. AB - INTRODUCTION: The inaugural meeting of the Safety Pharmacology Society (SPS) was in 2001, soon after ICH S7A had been adopted. The 10th anniversary is an appropriate milestone at which to analyse trends in the science and themes of safety pharmacology, as reflected in posters presented at the annual meetings. METHODS: The source information was the poster abstract booklets from each of the first ten annual meetings. RESULTS: The number of posters rose steadily from 34 in 2001 to 201 in 2010. The proportion of posters containing in vitro data has remained constant throughout the decade at ~30%. In terms of organ functions, themes relating to the cardiovascular system (CVS) have always generated the majority of posters, remaining above 60% of the total for the last 9years. The dominant theme has been around 'QT liability'. This peaked in 2003 at 68% of all posters presented, around the time of the ICHS7B discussions, and has remained above 30% thereafter. Apart from 2003 (dipping to 4%), CNS-related posters have remained steady at 11-17% throughout the decade. Respiratory-related posters have remained at 5-8% over the last 5years. Gastrointestinal (GI)-related posters have contributed 2-6% throughout the decade, and renal-related posters 1-3%. Posters on combined organ assessments have appeared in recent years. The relative emphasis on the different organ functions is broadly proportional to the causes of candidate drug attrition preclinically, whereas both CNS and GI are under represented when considering their contribution to significant adverse effects during clinical development. DISCUSSION: Trends are either regulatory-driven (e.g. increase in posters on abuse-dependence liability since EMEA/CHMP/SWP/94227/2004), technology-driven (e.g. automated hERG assay; left ventricular function; non-invasive CVS measurements; stem cells, etc.), or relate to the predictive ability of safety pharmacology data (e.g. clinical translation initiatives; concordance between in vitro and in vivo preclinical data; integrated risk assessment; PK-PD relationships, etc.). PMID- 21635958 TI - Regulation of LXR by fatty acids, insulin, growth hormone and tumor necrosis factor-alpha in rainbow trout myocytes. AB - The liver X receptor (LXR) has recently been described in salmonids. In mammals, this receptor is already known as a transcriptional factor that regulates diverse aspects of cholesterol, fatty acid and carbohydrate metabolism in various tissues, including muscle. Here we examined LXR in trout myocytes. For this purpose, we analyzed LXR target gene expression and gene profile during myocyte development. In addition, we studied the transcriptional regulation of LXR by hormones, a pro-inflammatory mediator and unsaturated fatty acids. Trout myocytes were incubated with LXR agonists (T091317, 22(R)-hydroxycholesterol) and unsaturated fatty acids for 24h. Furthermore, differentiated myocytes were incubated with insulin and growth hormone (GH) for 3h, 6h and 18h, and with tumor necrosis factor-alpha (TNFalpha) for 24h. Samples were also obtained in various stages of cell differentiation. Our results demonstrate that lipoprotein lipase (LPL), fatty acid synthase (FAS), ATP-binding cassette transporter A1 (ABCA1), peroxisome proliferator-activated receptor-alpha and beta (PPARalpha, beta) are target genes of LXR in trout muscle. LXR agonists increased LXR expression, thereby indicating that this receptor is autoregulated. Unsaturated fatty acids downregulated LXR gene expression. This observation suggests a regulatory mechanism of these molecules on LXR-mediated fatty acid synthesis and uptake. TNFalpha did not modulate LXR gene transcription. Expression of the LXR gene was activated by insulin and GH. These results suggest that LXR may play a lipogenic role through insulin stimulation and a tendency to promote anabolic effects through GH on trout myocytes. PMID- 21635959 TI - Elucidating the responses and role of the cardiovascular system in crocodilians during diving: fifty years on from the work of C.G. Wilber. AB - In 1960, C.G. Wilber in a laboratory-based study described for the first time the changes in heart rate with submergence in the American alligator noting in particular the marked bradycardia that occurred during forced dives. This short review summarizes the major advances in our understanding of diving and the responses and role of the cardiovascular system of crocodilians during submergence in the 50 years since Wilber published his findings. These advances are attributable in part to the technological advances made in physiological monitoring devices and wildlife telemetry that have not only provided greater elucidation of the hemodynamics of the unique crocodilian cardiovascular system but also allowed the natural diving behaviors and heart rates in free-ranging crocodiles to be recorded. Of note, telemetric field-based studies have revealed that wild free-ranging crocodiles typically undertake only short dives, less than 20 min, yet crocodiles are also capable of dives of many hours in duration. In contrast to Wilber's study, dives recorded from free-ranging crocodiles were found to be accompanied by only a modest bradycardia, highlighting the often confounding effects associated with captive animals monitored under laboratory conditions. More recent studies have also documented the complex central flow and pressure patterns of crocodilians, including a pulmonary to systemic shunt that can be initiated by a unique intracardiac valve located in the subpulmonary conus. The role and significance of this cardiac shunt remains controversial and the focus of recent lab-based studies. We contend that elucidation of the role and significance of the cardiac shunt in crocodilians will only be achieved by monitoring telemetrically the central cardiovascular flows and pressures in non captive animals that are undisturbed and free-ranging. This presents the challenge ahead in the next 50 years. PMID- 21635960 TI - Isolation and characterization of two strains of Fusarium oxysporum causing potato dry rot in Solanum tuberosum in Colombia. AB - BACKGROUND: Fusarium oxysporum has worldwide distribution and causes severe vascular wilt or root rot in many plants. Strains are classified into formae speciales based on their high degree of host specificity, of which multilocus sequence typing provides a fairly good estimate. AIMS: The main aim of this study was to identify the causal agent of an infected potato tuber in Colombia. METHODS: Two F. oxysporum isolates were recovered from a potato tuber showing symptoms of dry rot. Both macroscopic and microscopic morphology differences were observed between the two isolates. Koch's postulates were verified and in quantitative tuber pathogenecity trials, both isolates induced moderate dry rot. Ribosomal internal transcribed spacer (ITS) and partial intergenic spacer region (IGS) sequences were PCR-amplified, sequenced and shown to be identical for the two isolates. A maximum parsimony phylogeny was created using F. oxysporum IGS sequences available in the Genebank database, which does not include sequences from the formae speciales tuberosi. RESULTS: Our two isolates were most closely related to a red clover (Trifolium pratense) pathogenic isolate and two non pathogenic F. oxysporum isolates from birdsfoot trefoil (Lotus corniculatus) and Lycopersicon sp. rhyzosphere (99% identity). CONCLUSIONS: These experiments showed that our isolates are not restricted to potato and that a molecular marker is needed to differentiate the formae speciales since the IGS and EF-1alpha do not have the power to do it. PMID- 21635961 TI - Plant extracts to control Alternaria alternata in Murcott tangor fruits. AB - BACKGROUND: Alternaria alternata causes the Alternaria brown spot disease (ABS) in many tangerines and their hybrids worldwide. Plant extracts offer an alternative method for controlling this disease, which control is based on chemical fungicides. AIMS: To identify plant species with antifungal properties against A. alternata, the causal agent of the ABS. METHODS: Plant extracts prepared from leaves, barks, flowers, and stalks collected from 105 plant species in the State of Minas Gerais, Brazil, were tested for activity against the fungus A. alternata in vitro and in vivo. RESULTS: The most promising extract was obtained from Anadenanthera colubrina, which reduced the disease on Murcott tangor fruits to levels obtained with commercial fungicides. Artemisia annua, Cariniana estrelensis, Ficus carica, and Ruta graveolens presented moderate in vitro antifungal activity, but no effects were observed on the disease when the extracts were applied to fruits inoculated with the fungus. Besides, A. colubrina was the most active extract against A. alternata in the in vitro assay. CONCLUSIONS: The results obtained in the in vitro and in vivo assays suggested that the fungal growth test, which uses 96-well polypropylene plates, seems to be appropriate for selecting potential plant species for testing new methods to control ABS. PMID- 21635962 TI - [In vitro antifungal activity of ajoene on five clinical isolates of Histoplasma capsulatum var. capsulatum]. AB - BACKGROUND: The number of histoplasmosis cases have considerably increased since the advent of AIDS, and the therapy for this mycosis is not always effective, as well as having adverse effects. AIMS: To evaluate the inhibitory effect of ajoene on five clinical isolates of Histoplasma capsulatum, on the mycelial form, using Sabouraud dextrose broth (SDB) and RPMI-1640 culture media. METHODS: Growth curves and inhibitory activity of the drug (at concentrations of 1.25 ug/ml to 20 MUg/ml) were performed at room temperature, under mechanical agitation, and the turbidimetric readings (540 nm) were recorded every 48 h for 14 days, in both culture media. Generation times (GT) were calculated and graphs were constructed to estimate Minimal Inhibitory Concentrations (MIC) and Inhibitory Concentration 50% (IC(50)). The fungicidal minimal concentrations (FMC) were determined by plate cultures. The U-Mann-Whitney and t-test with a significance level of 0.05 were used to evaluate the statistical significance between culture media and GT, MIC, IC(50) MFC and fungistatic effect (FE). RESULTS: In both media and for all isolates, growth curves showed a GT of 43 to 67 hrs, an FE at 1.25-2.5 MUg/ml, and a MFC at 5-10 MUg/ml of ajoene. Values of MIC were 2.5-5 in SDB and in RPMI medium these values were 1.25-5 MUg/ml of ajoene. For IC(50), in SDB, the values were 1.9-2.6 ug/ml and in RPMI medium, they were of 3.8-4.3 MUg/ml of ajoene. There were no significance differences between culture media for GT, FE, MIC, IC(50) and MFC (p>0.05). CONCLUSIONS: These findings corroborate that ajoene inhibits the growth of the mycelial form of H. capsulatum. PMID- 21635963 TI - NADPH-oxidase but not inducible nitric oxide synthase contributes to resistance in a murine Staphylococcus aureus Newman pneumonia model. AB - Staphylococcus aureus is a pathogen that often causes severe nosocomial infections including pneumonia. The present study was designed to examine innate phagocyte mediated immune mechanisms using a previously described murine S. aureus Newman pneumonia model. We found that BALB/c mice represent a more susceptible mouse strain compared to C57BL/6 mice after intranasal S. aureus Newman challenge. Depletion experiments revealed that neutrophils are a crucial determinant for resistance whereas depletion of alveolar macrophages protected mice to some degree from acute pulmonary S. aureus challenge. C57BL/6 mice lacking the subunit gp91phox of the NADPH-oxidase (gp91phox-/- mice) proved to be highly susceptible against the pathogen. In contrast, C57BL/6 inducible nitric oxidase synthase deficient (iNOS-/-) mice did not differ in their clinical outcome after infection. Neither bone marrow macrophages from iNOS-/- nor from gp91phox-/- mice were impaired in controlling intracellular persistence of S. aureus. Our data suggest that neutrophil and NADPH-oxidase mediated mechanisms are essential components in protecting the host against pulmonary S. aureus Newman challenge. On contrary, macrophages as well as NO mediated mechanisms do not seem to play a critical role for resistance in this model. PMID- 21635965 TI - Macrolides: new therapeutic perspectives in lung diseases. AB - There is strong clinical evidence for the effectiveness of macrolides in the treatment of a number of chronic airway diseases through their immunomodulatory effects. Recently, new information has been released supporting the view that macrolides may also be beneficial in pathologic situations associated with altered repair of the alveolar structure, such as those observed in interstitial lung diseases and fibrosis. It is proposed that macrolides may contribute to lung regeneration through their actions on several components of the remodeling process. The present review provides new insights on the effects of macrolides on the regenerative response of alveolar epithelium to injury. It also discusses novel findings which suggest that macrolides may contribute to alveolar surfactant homeostasis. PMID- 21635966 TI - Decreased expression of protease-activated receptor 4 in human gastric cancer. AB - Protease-activated receptors (PARs) are a unique family of G-protein coupled receptors. PAR4, the most recently identified PAR member, was reported to be overexpressed during the progression of colon and prostate cancers. Though PAR4 mRNA was detected in normal stomach, the role of PAR4 in gastric cancer has not been investigated. In this study, differential expression of PAR4 was measured by real-time PCR (n=28) and tissue microarrays (n=74). We showed that PAR4 was located from basal to middle portions of normal gastric mucosa. PAR4 expression was remarkably decreased in gastric cancer tissues as compared with matched noncancerous tissues, especially in positive lymph node or low differentiation cancers. Furthermore, methylation of the PAR4 promoter in cell lines was assessed by treatment with 5-aza-2'-deoxycytidine and genomic bisulfite sequencing. AGS and N87 human gastric cancer cell lines did not express PAR4, as compared to HT 29 human colon cancer cell line with significant PAR4 expression. Treatment with 5-aza-2'-deoxycytidine restored PAR4 expression in AGS and N87 cells, which exhibited significantly more 5-methylcytosines in the PAR4 promoter compared with HT-29 cells. Our results revealed that down-regulation of PAR4 expression occurs frequently in gastric cancers and exhibits association with more aggressive gastric cancer. Interestingly, the loss of PAR4 expression in gastric cancers may result from hypermethylation of the PAR4 promoter. PMID- 21635964 TI - Inflammation in diabetic retinopathy. AB - Diabetes causes a number of metabolic and physiologic abnormalities in the retina, but which of these abnormalities contribute to recognized features of diabetic retinopathy (DR) is less clear. Many of the molecular and physiologic abnormalities that have been found to develop in the retina in diabetes are consistent with inflammation. Moreover, a number of anti-inflammatory therapies have been found to significantly inhibit development of different aspects of DR in animal models. Herein, we review the inflammatory mediators and their relationship to early and late DR, and discuss the potential of anti-inflammatory approaches to inhibit development of different stages of the retinopathy. We focus primarily on information derived from in vivo studies, supplementing with information from in vitro studies were important. PMID- 21635967 TI - A procedural framework for good imaging practice in pharmacological fMRI studies applied to drug development #1: processes and requirements. AB - There is increasing interest in the application of quantitative magnetic resonance imaging (MRI) methods to drug development, but as yet little standardization or best practice guidelines for its use in this context. Pharmaceutical trials are subject to regulatory constraints and sponsor company processes, including site qualification and expectations around study oversight, blinding, quality assurance and quality control (QA/QC), analysis and reporting of results. In this article, we review the processes on the sponsor side and also the procedures involved in data acquisition at the imaging site. We then propose summary recommendations to help guide appropriate imaging site qualification, as part of a framework of 'good imaging practice' for functional (f)MRI studies applied to drug development. PMID- 21635968 TI - Genetic and epigenetic biomarkers of colorectal cancer. AB - Cancer is a heterogeneous disease caused, in part, by genetic and epigenetic alterations. These changes have been explored in studies of the pathogenesis of colorectal cancer (CRC) and have led to the identification of many biomarkers of disease progression. However, the number of biomarkers that have been incorporated into clinical practice is surprisingly small. We review the genetic and epigenetic mechanisms of colorectal cancer and discuss molecular markers recommended for use in early detection, screening, diagnosis, determination of prognosis, and prediction of treatment outcomes. We also review important areas for future research. PMID- 21635969 TI - Repeat stool testing to diagnose Clostridium difficile infection using enzyme immunoassay does not increase diagnostic yield. AB - BACKGROUND & AIMS: Clostridium difficile infection (CDI) is a hospital-acquired infection with increasing incidence and severity. The most frequently used test to diagnose CDI is an enzyme immunoassay (EIA) for toxins A and B in stool samples. It is common to test 2 or more stool samples, based on the assumption that this detects CDI with greater sensitivity than analysis of 1 sample. We investigated whether repeat stool testing significantly improves the diagnostic yield for CDI. METHODS: We performed a retrospective analysis of hospitalized patients who were tested for CDI using EIA. From year 2005 to 2008, 39,402 stool samples from 17,971 patients with 29,373 diarrhea episodes were tested. Transition probabilities were calculated based on results from repeated tests. RESULTS: A total of 2692 diarrheal episodes (9.17%) were diagnosed with CDI. Based on results of 3 consecutive tests, 2675 (99.36%) were diagnosed with CDI. The first stool sample tested produced positive results for 90.7% of cases. When samples were tested consecutively, for the second and third time, an additional 6.6% and 2% patients had positive test results, respectively. If the first test result was negative, the probability of the second test result being positive was 2.7%. If the first 2 test results were negative, the probability of the third test result being positive was 2.3%. CONCLUSIONS: In patients who had multiple stool samples tested for CDI by EIA, almost 91% were accurately diagnosed based on the results of a single stool sample alone. Subsequent testing yielded a positive result in only 8.6% of patients. We therefore recommend that repeat testing not be done on a routine basis because it does not significantly improve diagnostic yield. PMID- 21635970 TI - Spatiotemporal phylogenetic analysis and molecular characterization of coxsackievirus A4. AB - Coxsackievirus A4 outbreaks occurred in Taiwan in 2004 and 2006. The spatiotemporal transmission of this error-prone RNA virus involves a continuous interaction between rapid sequence variation and natural selection. To elucidate the molecular characteristics of CV-A4 and the spatiotemporal dynamic changes in CV-A4 transmission, worldwide sequences of the 3' VP1 region (420 nt) obtained from GenBank were analyzed together with sequences isolated in Taiwan from 2002 to 2009. Sequences were characterized in terms of recombination, variability, and selection. Phylogenetic trees were constructed using neighbor-joining, maximum likelihood and Monte Carlo Markov Chain methods. Spatiotemporal dynamics of CV-A4 transmission were further estimated by a Bayesian statistical inference framework. No recombination was detected in the 420 nt region. The estimated evolution rate of CV-A4 was 8.65 * 10(-3) substitutions/site/year, and a purifying selection (d(N)/d(S)=0.032) was noted over the 3' VP1 region. All trees had similar topology: two genotypes (GI and GII), each including two subgenotypes (A and B), with the prototype and a Kenyan strain in separate branches. The results revealed that the virus first appeared in USA in 1950. Since 1998, it has evolved into the Kenya, GI-A (Asia) and GII-A (Asia and Europe) strains. Since 2004, GI-B and GII-B have evolved continuously and have remained prevalent. The co-existence of several positive selection lineages of GI-B in 2006 indicates that the subgenotype might have survived lineage extinction. This study revealed rapid lineage turnover of CV-A4 and the replacement of previously circulating strains by a new dominant variant. Therefore, continuous surveillance for further CV-A4 transmission is essential. PMID- 21635971 TI - The regulation of CD4+ T cell immune responses toward Th2 cell development by prostaglandin E2. AB - As an important immune mediator, PGE2 plays an important role in the immune tolerance, autoimmune diseases, immune regulation and tumor immunotolerance. PGE2 is considered to be a promising candidate for the control of the immune diseases. To further understand the immuno-modulating effects of PGE2 on CD4+ T cells, in vitro investigation was conducted in the present study. The results showed that PGE2 inhibited the proliferation of T cells in vitro in a dose-dependent manner. Gene expression profiling showed that 1716 genes were down regulated and 73 genes were up regulated with a change of 1.5 fold. Several signal transduction pathways were involved, such as TNF-alpha and NF-kB signaling pathway, T cell receptor signaling pathway, IL-2 signaling pathway, and MAPK pathway. The results showed that PGE2 inhibited IFN-gamma, TNF-alpha and IL-4 production by CD4+ T cells 24h after cell culture. A comparison between IFN-gamma and IL-4 production showed that PGE2 enhanced the relative ratio of IL-4 to IFN-gamma in CD4+ T cells culture, and regulated CD4+ T cells toward Th2 cell development. The results of the present study indicated that PGE2 has the potential to treat Th1-mediated inflammatory diseases by regulating CD4+ T cells toward Th2 cell immune response. PMID- 21635973 TI - Autoimmune hepatitis-like disease in C57BL/6 mice infected with mouse hepatitis virus A59. AB - Mouse hepatitis virus A59 (MHV A59) induces autoantibodies (autoAb) to fumarylacetoacetate hydrolase (FAH), a soluble cytosolic enzyme present in the liver and kidneys, in various mouse strains. The aim of this work was to amplify and diversify the autoimmune response restricted to FAH through the use of the exogenous adjuvant called PADRE. Accordingly, C57BL/6 mice were chosen, because these animals respond to PADRE better than other mouse strains. Results presented herein indicate that, surprisingly, C57BL/6 mice developed signs of autoimmune hepatitis-like disease (AIH), including transient hypergammaglobulinemia, elevated transaminases, autoAb directed against different liver proteins and hepatic cellular infiltrates, indicating that a new model of experimental AIH could be generated by a viral inoculation. Furthermore, PADRE administration amplified the MHV effect, extending the duration of hypergammaglobulinemia and increasing the binding of autoAb as well as the degree of hepatic infiltrates. However, the adjuvant did not expand the time of the symptoms. Additionally, since plasmatic uric acid and high-mobility group box protein 1 (HGMB1) concentrations augmented in MHV- and/or PADRE-treated mice, it is suggested that both alarmins were probably involved in the spreading of the immune response induced by the viral infection and the adjuvant administration. PMID- 21635972 TI - Resolving the identity myth: key markers of functional CD4+FoxP3+ regulatory T cells. AB - Authenticating markers for the functional suppressive CD4(+)FoxP3(+) regulatory T cells (Tregs) are important for the quantitative identification and enrichment of viable Tregs for possible therapeutic use. CD25 as a surrogate marker of Tregs has some limitations, which prompted investigators to identify more specific marker(s) of Tregs. The search for a firm molecular definition of Tregs resulted in the identification of FoxP3 as a better marker of this subset of CD4 cells. Nevertheless, FoxP3(+) Tregs are phenotypically and functionally heterogeneous. Even in normal mice, only a minority of FoxP3(+) T cells are potent suppressor cells. Therefore, additional marker(s) are required for delineation of truly functional Tregs. In this review, the studies identifying markers of functional Tregs, both in mouse and in human, and their functional implications are discussed. Our finding that TNFR2, which mediates the effect of TNF on the activation of Tregs, is a superb marker of the most suppressive subset of mouse Tregs and its application in the identification of functional human Tregs will also be reviewed. PMID- 21635974 TI - The implications of mitochondrial DNA copy number regulation during embryogenesis. AB - Mutations of mitochondrial DNA (mtDNA) cause a wide array of multisystem disorders, particularly affecting organs with high energy demands. Typically only a proportion of the total mtDNA content is mutated (heteroplasmy), and high percentage levels of mutant mtDNA are associated with a more severe clinical phenotype. MtDNA is inherited maternally and the heteroplasmy level in each one of the offspring is often very different to that found in the mother. The mitochondrial genetic bottleneck hypothesis was first proposed as the explanation for these observations over 20 years ago. Although the precise bottleneck mechanism is still hotly debated, the regulation of cellular mtDNA content is a key issue. Here we review current understanding of the factors regulating the amount of mtDNA within cells and discuss the relevance of these findings to our understanding of the inheritance of mtDNA heteroplasmy. PMID- 21635975 TI - Enzymatic characterization of a serralysin-like metalloprotease from the entomopathogen bacterium, Xenorhabdus. AB - We investigated the enzymatic properties of a serralysin-type metalloenzyme, provisionally named as protease B, which is secreted by Xenorhabdus bacterium, and probably is the ortholog of PrA peptidase of Photorhabdus bacterium. Testing the activity on twenty-two oligopeptide substrates we found that protease B requires at least three amino acids N-terminal to the scissile bond for detectable hydrolysis. On such substrate protease B was clearly specific for positively charged residues (Arg and Lys) at the P1 substrate position and was rather permissive in the others. Interestingly however, it preferred Ser at P1 in the oligopeptide substrate which contained amino acids also C-terminal to the scissile bond, and was cleaved with the highest k(cat)/K(M) value. The pH profile of activity, similarly to other serralysins, has a wide peak with high values between pH 6.5 and 8.0. The activity was slightly increased by Cu(2+) and Co(2+) ions, it was not sensitive for serine protease inhibitors, but it was inhibited by 1,10-phenanthroline, features shared by many Zn-metalloproteases. At the same time, EDTA inhibited the activity only partially even either after long incubation or in excess amount, and Zn(2+) was inhibitory (both are unusual among serralysins). The 1,10-phenanthroline inhibited activity could be restored with the addition of Mn(2+), Cu(2+) and Co(2+) up to 90-200% of its original value, while Zn(2+) was inefficient. We propose that both the Zn inhibition of protease B activity and its resistance to EDTA inhibition might be caused by an Asp in position 191 where most of the serralysins contain Asn. PMID- 21635976 TI - Thrombocytopenia-absent radius (TAR) syndrome: a clinical genetic series of 14 further cases. impact of the associated 1q21.1 deletion on the genetic counselling. AB - Thrombocytopenia-absent radius Syndrome (TAR) is a rare congenital malformation syndrome of complicated transmission. 1q21.1 deletion is necessary but not sufficient for its expression. We report the result of a French multicentric clinical study, and we emphasized on the role of the associated 1q21.1 deletion in the diagnosis and the genetic counselling of our patients. We gathered information on 14 patients presenting with TAR syndrome and referred for genetic counselling in six different university hospitals (8 foetuses, 1 child and 5 adults). Clinical or pathology details, as well as skeletal X-rays were analyzed. Genetic studies were performed by Array-CGH, and Quantitative Multiplex PCR. We demonstrated the very variable phenotypes of TAR syndrome. Female:male ratio was ~2:1. All patients presented with bilateral radial aplasia/hypoplasia with preserved thumbs. Phocomelia and lower limb anomalies were present in 28% of the cases. We reported the first case of cystic hygroma on affected foetus. 1q21.1 deletions ranging from 330 to 1100 kb were identified in all affected patients. Most of them were inherited from one healthy parent (80%). The identification of a 1q21.1 deletion allowed confirmation of TAR syndrome diagnosis, particularly in foetuses and in atypical phenotypes. Additionally, it allowed accurate genetic counselling, especially when it occurred de novo. These findings allowed discussing the diagnostic criteria and management towards TAR syndrome. PMID- 21635977 TI - Protein identification using MS/MS data. AB - The subject of this tutorial is protein identification and characterisation by database searching of MS/MS Data. Peptide Mass Fingerprinting is excluded because it is covered in a separate tutorial. Practical aspects of database searching are emphasised, such as choice of sequence database, effect of mass tolerance, and how to identify post-translational modifications. The relationship between sensitivity and specificity is discussed, as is the challenge of using peptide match information to infer which proteins were present in the sample. Since these tutorials are introductory in nature, most references are to reviews, rather than primary research papers. Some familiarity with mass spectrometry and protein chemistry is assumed. There is an accompanying slide presentation, including speaker notes, and a collection of web-based, practical exercises, designed to reinforce key points. This Tutorial is part of the International Proteomics Tutorial Programme (IPTP 6). PMID- 21635978 TI - Identification and validation of six proteins as marker for endemic nephropathy. AB - Endemic nephropathy (EN) is defined as a slow progressive renal tubulointestitial disease that mainly occurs in the restricted areas of the Balkan Peninsula. The complexity of the pathogenesis of EN makes its earlier diagnosis very difficult. Urine samples from healthy volunteers from EN regions, EN patients with proteinuria less than 150 mg/L and EN patients with proteinuria more than 150 mg/L, patients with acute kidney injury, patients with diabetic nephropathy and healthy volunteers from Germany were collected. The urinary proteome analyses were performed using 2-D DIGE and mass spectrometry. The validation of biomarkers was investigated by two approaches (Western blot (WB) and dot blot) in successively increasing size - and partially overlapping - sample sets. Comparative and statistical analyses of the proteomics data from the different patient groups allowed the identification of six proteins (alpha-1-microglobulin, alpha-2-glycoprotein-1, beta-2-microglobulin, mannose-binding-lectin-2, protection-of-telomeres-protein-1, and superoxide-dismutase [Cu-Zn]), which were able to discriminate EN with low and high proteinuria from the other groups with high significance (p<0.05). The reliability of the identified proteins as EN marker was underlined with high statistical significance using WB analyses (sensitivity 66.7-98% and specificity 70-100%), whereas the dot blot analyses revealed a decrease in the sensitivity and specificity of these biomarkers. PMID- 21635979 TI - Proteomic characterization of thymocyte-derived microvesicles and apoptotic bodies in BALB/c mice. AB - Several studies have characterized exosomes derived from different cell sources. In this work we set the goal of proteomic characterization of two less studied populations of membrane vesicles, microvesicles (100-800 nm) and apoptotic bodies (> 800 nm) released by thymus cells of BALB/c mice. The vesicles were isolated by the combination of differential centrifugation and gravity driven multistep filtration of the supernatant of thymus cell cultures. The size distribution of vesicle preparations was determined by transmission electron microscopy. Proteins were released from the vesicles, digested in solution, and analyzed using nano HPLC/MS(MS). Ingenuity pathway analysis was used to identify functions related to membrane vesicle proteins. In apoptotic bodies and microvesicles we have identified 142 and 195 proteins, respectively. A striking overlap was detected between the proteomic compositions of the two subcellular structures as 108 proteins were detected in both preparations. Identified proteins included autoantigens implicated in human autoimmune diseases, key regulators of T-cell activation, molecules involved in known immune functions or in leukocyte rolling and transendothelial transmigration. The presence and abundance of proteins with high immunological relevance within thymocyte-derived apoptotic bodies and microvesicles raise the possibility that these subcellular structures may substantially modulate T-cell maturation processes within the thymus. PMID- 21635980 TI - Shotgun proteomic analysis of the unicellular alga Ostreococcus tauri. AB - Ostreococcus tauri is a unicellular green alga and amongst the smallest and simplest free-living eukaryotes. The O. tauri genome sequence was determined in 2006. Molecular, physiological and taxonomic data that has been generated since then highlight its potential as a simple model species for algae and plants. However, its proteome remains largely unexplored. This paper describes the global proteomic study of O. tauri, using mass spectrometry-based approaches: phosphopeptide enrichment, cellular fractionation, label-free quantification and (15)N metabolic labeling. The O. tauri proteome was analyzed under the following conditions: sampling at different times during the circadian cycle, after 24h of illumination, after 24h of darkness and under various nitrogen source supply levels. Cell cycle related proteins such as dynamin and kinesin were significantly up-regulated during the daylight-to-darkness transition. This is reflected by their higher intensity at ZT13 and this transition phase coincides with the end of mitosis. Proteins involved in several metabolic mechanisms were found to be up-regulated under low nitrogen conditions, including carbon storage pathways, glycolysis, phosphate transport, and the synthesis of inorganic polyphosphates. Ostreococcus tauri responds to low nitrogen conditions by reducing its nitrogen assimilation machinery which suggests an atypical adaptation mechanism for coping with a nutrient-limited environment. PMID- 21635981 TI - Calcium supplementation attenuates citrate-related changes in bone metabolism: a placebo-controlled crossover study in healthy volunteers. AB - BACKGROUND: Citrate is the anticoagulation of choice in apheresis procedures. Citrate anticoagulation results in a short-term increase in serological markers of bone turnover, with uncertain clinical significance. AIM: To understand the effect of calcium supplementation on serological bone turnover markers during an acute citrate load as a mimic of citrate anticoagulation during apheresis procedures. METHODS: A placebo-controlled, crossover study was conducted in 22 healthy volunteers. Volunteers received a standardized citrate load at a fixed dose of 1.5 mg/kg of body weight/min for 80 min for three times and a single placebo infusion as a control. Each intervention was separated by a wash-out interval of 2 to 3 weeks. During two citrate infusions, volunteers received an additional calcium supplementation, consisting of either oral administration of calcium carbonate or an i.v. bypass infusion of calcium gluconate. Serial blood samples were collected for the determination of ionized calcium (iCa), intact parathyroid hormone (iPTH) and markers of bone remodeling, C-telopeptide of type 1 collagen (CTX) and osteocalcin (OC). RESULTS: The infusion of citrate without calcium supplementation resulted in an increase in the bone formation marker OC and the bone resorption marker CTX, in addition to the changes in iPTH and iCa. The administration of calcium by either oral administration or as an i.v. bypass infusion attenuated the observed changes in CTX, but showed no effects on the elevation of the bone formation marker OC. There was no difference in the attenuation of CTX between the two calcium formulations. However, the i.v. application of calcium gluconate had a superior effect in reducing the change of serum iPTH and iCa as compared to the oral administration of calcium carbonate. CONCLUSIONS: Calcium supplementation is an effective method in damping the citrate-related transient increase of the serological bone resorption marker CTX. As a mimic for the citrate-based apheresis procedure, our data may enforce the prophylactic application of calcium supplementation to attenuate the short-term elevation of bone resorption related to an acute citrate load. PMID- 21635984 TI - Angioaccess for hemodialysis. PMID- 21635985 TI - Foreword. Continued debate regarding optimal surgery, radiation techniques, and chemotherapy regimens. PMID- 21635986 TI - Gastric cancer. PMID- 21635987 TI - Cardiovascular implications of antihyperglycemic therapies for type 2 diabetes. AB - BACKGROUND: Several risk factors for cardiovascular disease (CVD), including insulin resistance/hyperinsulinemia, hyperglycemia, overweight/obesity, dyslipidemia, and hypertension, are often present in varying combinations in patients with type 2 diabetes mellitus (DM). Patients with a clustering of these risk factors, termed the metabolic syndrome, are at greater risk for CVD than are patients with only a single risk factor. Although glycemic control is the central feature of type 2 DM management, patients require an individualized approach to therapy that takes their other CVD risk factors into account. OBJECTIVE: This review examined the effects of antidiabetes therapy on glycemic control, as well as its potential to affect body weight, serum lipids, and blood pressure (BP), and thus CVD risk. METHODS: Information was obtained by searching the MEDLINE and EMBASE databases from 1995 through March 2010. The search terms included type 2 DM, metabolic syndrome, CV complications of type 2 DM, and therapy for type 2 DM. Articles that described relevant details of the metabolic syndrome, CV complications of type 2 DM, and effects of antidiabetes therapy on glycosylated hemoglobin, body weight, serum lipids, and BP were selected for in-depth review. Only English language publications were reviewed. Clinical trials, meta-analyses, and review articles on the key words were preferentially selected for review and analysis. Non-English language publications, case reports, letters to the editor, and similar types of publications were excluded. RESULTS: Although all approved antidiabetes agents lowered glucose, their effect on other CV risk factors, such as BP, lipids, and weight, differed significantly. Therapy with insulin, the sulfonylureas, and the thiazolidinediones was associated with weight gain. Metformin and the dipeptidyl-peptidase-4 inhibitors were generally considered weight neutral, whereas the glucagon-like peptide-1 receptor agonists and amylin agonists were associated with weight loss. Metformin, sulfonylureas, thiazolidinedioness, and dipeptidyl-peptidase-4 inhibitors had modest effects on serum lipid levels and BP. The glucagon-like peptide-1 receptor agonists generally had beneficial effects on serum lipid levels and systolic and diastolic BP. CONCLUSION: A wide variety of agents were available to aid glycemic control in patients with type 2 DM. These agents had variable effects on known CV risk factors that might be present in this patient population, including excess body weight, elevated BP, and increased serum lipids. Some of the newer agents improved glycemic control while also having potentially favorable effects on these CV risk factors. The impact of various agents on known CV risk factors should be considered when selecting a therapeutic regimen. PMID- 21635988 TI - Evaluating treatment algorithms for the management of patients with type 2 diabetes mellitus: a perspective on the definition of treatment success. AB - BACKGROUND: Traditional treatment of type 2 diabetes mellitus (T2DM) has focused on correcting hyperglycemia. However, T2DM is often accompanied by other conditions and risk factors, including hypertension, overweight/obesity, and dyslipidemia, that affect morbidity and mortality. A broader view toward treating the array of physiologic derangements may provide significant long-term outcomes benefits. OBJECTIVE: This perspective paper reviews recent data regarding the pathophysiology of T2DM, evaluates current treatment recommendations/algorithms, and discusses potential risks and benefits associated with the various therapeutic options and their combinations. METHODS: Information was obtained by a search of the PubMed and Embase databases using the key words type 2 diabetes mellitus, glycosylated hemoglobin, pathophysiology of type 2 diabetes, glycemic control, antidiabetes therapy, multifactorial therapy, and treatment algorithms for the period of 1985 to 2010. A representative number of relevant articles dealing with these topics was then selected for review. RESULTS: Three recently proposed treatment algorithms, the American Diabetes Association/European Association for the Study of Diabetes less well-validated algorithm, the American Association of Clinical Endocrinologists/American College of Endocrinology algorithm, and the DeFronzo algorithm, were compared and contrasted. Metformin is usually the first oral agent to be used when not contraindicated because of its ability to suppress hepatic glucose production. Some recommended agents, such as sulfonylureas, drive beta-cell failure even as they improve glycosylated hemoglobin. Others, such as glucagon-like peptide-1 (GLP-1) receptor agonists and thiazolidinediones, support and enhance beta-cell function. Also, some agents predispose to weight gain (eg, sulfonylureas and insulin), whereas others are weight neutral (eg, dipeptidyl-peptidase-4 inhibitors) or result in weight loss (eg, GLP-1 receptor agonists, pramlintide). Finally, the impact of these agents on associated cardiovascular risk factors is varied. Agents that improve glycemic control while favorably affecting blood lipid levels and/or systemic blood pressure may lead to improvements in morbidity and mortality. Attention to the mechanisms of action and associated consequences of the numerous pharmacologic treatment choices may provide clinicians a better selection of agents in treating patients with T2DM. CONCLUSIONS: Physicians should evaluate the array of treatment options available for patients with T2DM. An aggressive regimen including metformin, a thiazolidinedione, and a GLP-1 receptor agonist may improve insulin sensitivity and enhance beta-cell function. Addressing the pathophysiologic defects associated with T2DM, as well as the various associated cardiovascular risk factors, with combination therapy may slow the natural progression of the disease and development of its associated complications. PMID- 21635989 TI - Emerging antiplatelet therapies in percutaneous coronary intervention: a focus on prasugrel. AB - BACKGROUND: Prasugrel is the most recent addition to the available thienopyridine antiplatelet agents used to prevent ischemic events in patients with acute coronary syndrome undergoing percutaneous coronary intervention. OBJECTIVE: The aim of this article was to review published data on the efficacy and safety profile of prasugrel, cost considerations with its use, and its place in clinical care. METHODS: We searched PubMed and Ovid databases for English language clinical trial articles, published through December 2010, involving the use of prasugrel in human subjects. The key word prasugrel was used. The review focused on clinical trials, but other articles and Food and Drug Administration documents were also reviewed for relevant information. RESULTS: Phase II studies showed that prasugrel had a more powerful antiplatelet effect and was more effective in its inhibition of platelet activation than clopidogrel. In the only Phase III trial completed before its Food and Drug Administration approval, prasugrel demonstrated a decrease in the primary composite efficacy end point of the rate of cardiovascular death, nonfatal myocardial infarction, or nonfatal stroke relative to clopidogrel (643 [9.9%] vs 781 [12.1%], respectively; P = 0.001). Prasugrel was associated with a significantly higher risk of bleeding compared with clopidogrel. Non-coronary artery bypass graft-related thrombolysis in myocardial infarction (TIMI) major bleeding occurred in 146 (2.4%) patients in the prasugrel group versus 111 (1.8%) patients in the clopidogrel group (P = 0.03). For every 1000 patients treated with prasugrel instead of clopidogrel, a total of 23 myocardial infarctions could be prevented at the cost of 6 additional TIMI major bleeding events. However, this benefit was diminished with longer-term therapy. Adverse outcomes of prasugrel use outweighed its benefits in certain subgroups, including patients >75 years old, those weighing <60 kg, and patients with a history of stroke or transient ischemic attack. CONCLUSIONS: Prasugrel's clinical benefits were counterbalanced by an increase in bleeding risk compared with conventional thienopyridine treatment with clopidogrel. Current practice guidelines incorporated prasugrel as a treatment option, but at this time do not recommend that prasugrel be selected over clopidogrel in any patient subgroup. Further study is required to determine optimal dosing and proper patient selection with prasugrel treatment. PMID- 21635990 TI - Palonosetron versus other 5-HT(3) receptor antagonists for prevention of chemotherapy-induced nausea and vomiting in patients with cancer on chemotherapy in a hospital outpatient setting. AB - BACKGROUND: Despite favorable evidence from clinical trials for single-dose palonosetron versus other commercially available 5-HT(3)-receptor antagonists for the prophylaxis of chemotherapy-induced nausea and vomiting (CINV), clinical comparative data are scarce from hospital outpatient settings, where these antiemetic agents are used in patients diagnosed with cancer who are receiving chemotherapy (CTH). OBJECTIVE: The purpose of our retrospective study was to assess the hospital claims to evaluate the rate of uncontrolled CINV with antiemetic prophylaxis using palonosetron versus other 5-HT(3)-receptor antagonists in patients diagnosed with cancer who are receiving CTH (highly emetogenic CTH, moderately emetogenic CTH, low-emetogenic CTH, or minimally emetogenic CTH) treatment in a hospital outpatient setting. METHODS: Patients aged >=18 years who had cancer and were being treated with CTH and antiemetic prophylaxis with palonosetron (Group 1) and other 5-HT(3) receptor antagonists (Group 2) for the first time between April 1, 2007, and March 31, 2009, were identified using a hospital-service database. Within each CTH cycle, CINV events were identified through International Classification of Diseases (ICD)-9 codes for nausea, vomiting, and/or volume depletion (from Day 1 of each CTH administration until the end of the CTH cycle) or for use of rescue medications (Day 2 until the end of the CTH cycle). A multivariate regression model was developed to predict uncontrolled CINV event rates per CTH cycle between Groups 1 and 2 matched on CTH emetogenicity distribution in the study follow-up period (first of 8 cycles or 6 months). A subgroup analysis of patients on CTH with the highest risk of nausea and vomiting (highly emetogenic CTH or moderately emetogenic CTH) was also conducted. RESULTS: Of 9144 identified patients, 1775 were prescribed palonosetron (Group 1). Group 1 patients were statistically younger (61.2 vs 62.8 years; P < 0.001), composed of more females (57.1% vs 51.9%; P < 0.001) and more whites (72.8% vs 71.4%; all races P < 0.001), received more highly emetogenic CTH treatments (43.3% vs 28.5%; all CTH P < 0.001), and had more lung (26.1% vs 22.4%) and breast cancer patients (19.3% vs 15.3%; all cancer P < 0.001). The regression model predicted a 13.7% decrease in CINV event rate per CTH cycle for Group 1 versus Group 2. For Subgroup 1, the model predicted a 12.5% decrease in the CINV event rate per cycle in Group 1 patients versus those in Group 2. CONCLUSIONS: In this study, patients with cancer who were treated with CTH and on antiemetic prophylaxis using palonosetron were found to have significantly lower CINV event rates than those receiving other 5-HT(3) receptor antagonists. PMID- 21635991 TI - Hyperkalemia and renal function during monotherapy and dual renin-angiotensin blockade in the community setting. AB - BACKGROUND: In controlled trials, dual therapy with angiotensin-converting enzyme inhibitors (ACE-I) and angiotensin receptor blockers (ARBs) is associated with hyperkalemia and decreased renal function, but there is no information about these adverse effects in clinical practice. OBJECTIVE: The aim of this study was to assess the incidence of hyperkalemia and decreased renal function during dual therapy (ACE-I plus ARB) in a community-based setting. METHODS: In a retrospective cohort database study, we identified patients who received ARBs added to ongoing ACE-I therapy and who had at least 1 measurement of serum creatinine and potassium during each treatment period. We compared rates of hyperkalemia (>5.5 mmol/L) during equal periods of monotherapy and dual therapy and the rate of a significant rise in serum creatinine (>=0.5 mg/dL) between study periods. We assessed the impact of potential confounders on outcomes by logistic regression analysis. RESULTS: Among 425 patients (median follow-up 19 months for each treatment period), hyperkalemia was 2-fold more common during dual therapy than monotherapy (11.1% and 5.6% of patients, respectively) (relative risk = 1.96; 95% CI, 1.22-3.14; P < 0.001). In 77 patients with reduced renal function on monotherapy (serum creatinine >=1.5 mg/dL), the rate of hyperkalemia was 20.8/100 patient-years, resulting in a number needed to harm of 10.1 patients, compared with 52.6 patients among those with preserved renal function. Mean serum creatinine between treatment periods increased >0.5 mg/dL in 7.5% of patients, more commonly in patients with decreased (18.2%) than with preserved (5.2%) baseline renal function (P < 0.001). CONCLUSION: In the community setting, dual therapy was associated with hyperkalemia and a decrease in renal function. The absolute risks were especially high among patients with reduced baseline renal function. PMID- 21635992 TI - Effects of varenicline in adult smokers: a multinational, 24-week, randomized, double-blind, placebo-controlled study. AB - BACKGROUND: Prevalence rates of smoking are rising in developing countries. Previous trials evaluating the efficacy and tolerability of the smoking-cessation medication varenicline have used largely participants of Caucasian origin. OBJECTIVE: This study was conducted to evaluate the efficacy and tolerability of varenicline in populations of participants from Latin America, Africa, and the Middle East to investigate potential differences in the therapeutic response to varenicline. METHODS: This multinational, randomized, double-blind, placebo controlled trial was conducted at 42 centers in 11 countries (Latin America: Brazil, Colombia, Costa Rica, Mexico, and Venezuela; Africa: Egypt and South Africa; Middle East: Jordan, Lebanon, Saudi Arabia, and the United Arab Emirates). Participants were male and female smokers aged 18 to 75 years who were motivated to stop smoking; smoked >=10 cigarettes/d, with no cumulative period of abstinence >3 months in the previous year; and who had no serious or unstable disease within the previous 6 months. Subjects were randomized in a 2:1 ratio to receive varenicline 1 mg or placebo, BID for 12 weeks, with a 12-week nontreatment follow-up. Brief smoking-cessation counseling was provided. The main outcome measures were carbon monoxide-confirmed continuous abstinence rate (CAR) at weeks 9 to 12 and weeks 9 to 24. Adverse events (AEs) were recorded for tolerability assessment. RESULTS: Overall, 588 subjects (varenicline, 390; placebo, 198) were randomized and treated. The mean (SD) ages of subjects in the varenicline and placebo groups were 43.1 (10.8) and 43.9 (10.8) years, respectively; 57.7% and 65.7% were male; and the mean (SD) weights were 75.0 (16.0) and 76.7 (16.3) kg (range, 40.0-130.0 and 45.6-126.0 kg). CAR at weeks 9 to 12 was significantly higher with varenicline than with placebo (53.59% vs 18.69%; odds ratio [OR] = 5.76; 95% CI, 3.74-8.88; P < 0.0001), and this rate was maintained during weeks 9 to 24 (39.74% vs 13.13%; OR = 4.78; 95% CI, 2.97-7.68; P < 0.0001). Nausea, headache, and insomnia were the most commonly reported AEs with varenicline and were reported numerically more frequently in the varenicline group compared with the placebo group. Serious AEs (SAEs) were reported in 2.8% of varenicline recipients compared with 1.0% in the placebo group, with 6 subjects reporting psychiatric SAEs compared with none in the placebo group. CONCLUSION: Based on these data, varenicline was apparently efficacious and generally well tolerated as a smoking-cessation aid in smokers from selected sites in Latin America, Africa, and the Middle East. ClinicalTrials.gov identifier: NCT00594204. PMID- 21635993 TI - A better mousetrap. PMID- 21635994 TI - Cost-effectiveness of targeted therapy with cetuximab in patients with K-ras wild type colorectal cancer presenting with initially unresectable metastases limited to the liver in a German setting. AB - BACKGROUND: In patients with metastases limited to the liver (liver-limited disease [LLD]), effective therapies such as monoclonal antibodies combined with chemotherapy may facilitate metastasis resection and improve long-term survival. OBJECTIVE: This study assessed the cost-effectiveness of bevacizumab and cetuximab in the treatment of patients with colorectal cancer presenting with initially unresectable liver metastases of the Kirsten rat sarcoma viral oncogene homolog (K-ras) wild type, from the perspective of German statutory health insurance. METHODS: The health-economic modeling approach presented here made indirect comparisons between available data on bevacizumab and cetuximab treatment outcomes using evidence synthesis techniques, extrapolating from the follow-up duration of identified clinical trials to a longer time horizon of up to 10 years and inferring costs and health outcomes based on modeled patient pathways. Expert opinion and Delphi panel methods were used for some assumptions, when evidence was missing. Probabilistic sensitivity analyses and different scenario analyses were applied to test for uncertainty around input parameters and assumptions. RESULTS: For the metastatic colorectal cancer LLD population with K-ras wild-type genotype, mean overall survival estimates were 37.7 months for first-line treatment with cetuximab plus FOLFIRI (irinotecan, leucovorin, fluorouracil) and 30.4 months for bevacizumab plus FOLFOX (oxaliplatin, leucovorin, fluorouracil). Corresponding discounted survival estimates were 2.88 life-years with cetuximab plus FOLFIRI versus 2.38 life-years with bevacizumab plus FOLFOX, an average gain of 0.50 discounted life-years. The incremental cost effectiveness ratio of cetuximab plus FOLFIRI versus bevacizumab plus FOLFOX was ?15,020 (year 2010 ?) per life-year gained in the base case (with a 95% CI from the probabilistic sensitivity analysis of ?3806-?24,660). Results were robust in different scenario analyses as well as in the probabilistic sensitivity analysis. CONCLUSIONS: First-line treatment with cetuximab plus FOLFIRI offers a cost effective treatment option versus bevacizumab plus FOLFOX for the metastatic colorectal cancer LLD population with K-ras wild-type genotype in Germany. K-ras testing should be performed on all presenting cases of metastatic colorectal cancer to ensure access to this treatment option. PMID- 21635996 TI - Guns for EMS? Infrared guns can help assess & save lives. PMID- 21635995 TI - Comparative bioavailability of 2 tablet formulations of levodopa/benserazide in healthy, fasting volunteers: a single-dose, randomized-sequence, open-label crossover study. AB - BACKGROUND: Currently, levodopa administered with decarboxylase inhibitors is the gold standard for the management of the motor symptoms of Parkinson's disease, a neurodegenerative disorder characterized by the degeneration of dopaminergic neurons in the substantia nigra pars compacta. In Argentina, only 1 commercial product is available with such composition; this study was contracted by the manufacturer to comply with new generic product regulations. OBJECTIVE: The aim of this study was to evaluate the fasting bioavailability of a new generic formulation of levodopa 200 mg/benserazide 50 mg tablets (test) and compare this generic formulation with the branded formulation (reference) to meet regulatory criteria for marketing the test product in Argentina. METHODS: A randomized sequence, open-label, 2-period, crossover study was conducted between August and October 2009 in healthy Caucasian volunteers (n = 24; 18 males, aged 21 to 42 years, with a body mass index ranging from 19.7 to 26.0 kg/m(2)) in the fasted state. A single oral dose of the test or reference formulation was administered, and after a 7-day washout period, the other formulation was given. Blood samples were collected at baseline and at 10, 20, 30, 40, 50, 60, 70, 80, 90, and 105 minutes and 2, 2.5, 3, 3.5, 4, and 6 hours after dosing. Levodopa plasma concentrations were measured by high-performance liquid chromatography with electrochemical detection, without stereo-specificity assessment. The formulations were considered bioequivalent if the 90% CI of the geometric mean ratios (test/reference) for the C(max) and AUC(0-t) of levodopa were within the 0.8 to 1.25 range. Adverse events were monitored throughout the study, based on clinical parameters and patient reports. RESULTS: The geometric means (90% CI) of the C(max) for the test and reference formulations were 2462.02 (2312.06-3492.40) and 2542.85 (2394.49-3231.29) ng/mL, respectively; the AUC(0-t) was 3878.04 (3623.88-5393.09) and 3972.10 (3765.88-5393.02) ng/mL/h, respectively; and the AUC(0-infinity)was 4610.37 (4315.71-6315.70) and 4728.96 (4502.17-6828.26) ng/mL/h, respectively. There were no significant differences in pharmacokinetic parameters between the 2 formulations. The test:reference ratios for C(max), AUC(0-t), and AUC(0-infinity) were 96.82% (90% CI, 83.87-111.77), 97.63% (90% CI, 85.95-110.91), and 97.49% (90% CI, 84.09-113.02), respectively. No clinically significant adverse events were reported; this finding is probably the result of subjects not believing that their side effects were severe enough to be reported and not because of a genuine and absolute lack of predictable side effects. CONCLUSIONS: In this single-dose study, the test formulation of levodopa/benserazide tablets met the Argentinean criterion for bioequivalence to the reference formulation. (www.clinicaltrials.gov: NCT01327261). PMID- 21635997 TI - Making sense of shock. PMID- 21635998 TI - Making sense of shock. PMID- 21636000 TI - EMS providers who drive like maniacs. PMID- 21636001 TI - EMS providers who drive like maniacs. PMID- 21636002 TI - Can you keep a secret? Leaking information. PMID- 21636003 TI - Suction skills: be positive about negative pressure. PMID- 21636004 TI - Find your farmhouse: caring for the rest of our lives. PMID- 21636005 TI - More than skin deep: use caution when treating impalement injuries. PMID- 21636007 TI - Achieving 20/20 glottic visualization: clinical research provides lessons for a perfect view during intubation. PMID- 21636008 TI - New directions: MCHD redesign takes a forward-thinking approach. Interview by A J Heightman. PMID- 21636009 TI - Simulation station: Chicago Fire Department takes training to a new level. PMID- 21636010 TI - Weight loss is coupled with improvements to affective state in obese participants engaged in behavior change therapy based on incremental, self-selected "small changes". AB - The aim of this study was to investigate the effects of a group behavior change intervention involving self-selected, contextualized, and mediated goal setting on anthropometric, affective, and dietary markers of health. It was hypothesized that the intervention would elicit changes consistent with accepted health recommendations for obese individuals. A rolling program of 12-week "Small Changes" interventions during 24 months recruited 71 participants; each program accommodated 10 to 13 adults (body mass index [BMI] >= 30 kg/m2). Fifty-eight participants completed Small Changes. Repeated measures were made at baseline, 6 and 12 weeks. Anthropometric measures included height and weight (to calculate BMI), body composition, waist circumference, and blood pressure. Affective state was monitored using relevant validated questionnaires. Dietary assessment used 3 day household measures food diaries with Schofield equations to monitor underreporting. Relevant blood measures were recorded throughout. Across the measurement period, Small Changes elicited a significant reduction in body weight (baseline, 102.95 +/- 15.47 vs 12 weeks 100.09 +/- 16.01 kg, P < .0005), coupled with associated significant improvements in BMI, body fat percentage, and waist circumference measures. There were additional significant positive changes in measures of affective state including general well-being (baseline, 58.92 +/- 21.22 vs 12 weeks 78.04 +/- 14.60, P < .0005) and total mood disturbance (baseline, 31.19 +/- 34.03 vs 12 weeks 2.67 +/- 24.96, P < .0005). Dietary changes that occurred were largely consistent with evidenced-based recommendations for weight management and included significant reductions in total energy intake and in fat and saturated fat as a proportion of energy. The Small Changes approach can elicit a range of health-orientated benefits for obese participants, and although further work is needed to ascertain the longevity of such effects, the outcomes from Small Changes are likely to help inform health professionals when framing the future of weight management. Long-term follow-up of Small Changes is warranted. PMID- 21636011 TI - Intake of added sugars is not associated with weight measures in children 6 to 18 years: National Health and Nutrition Examination Surveys 2003-2006. AB - Studies examining an association between consumption of added sugars (AS) and weight measures in children are inconclusive. This study examined the association between intake of AS and 5 measures of weight or adiposity using a nationally recent representative sample of children. National Health and Nutrition Examination Surveys 2003-2006 24-hour recall data from children 6 to 18 years (n = 3136) were used. United States Department of Agriculture's definition of AS and MyPyramid Equivalents Database were used to estimate daily intake. Multiple linear regression and squared partial correlation coefficients were used to estimate the strength of association between weight, body mass index (BMI), BMI Z scores, waist circumference, and triceps and subscapular skinfolds as dependent variables with AS as the independent variable. Covariates were age, sex, race/ethnicity, poverty income ratio, total energy intake (kJ), and physical activity. Mean intake of AS was 23 +/- 0.55 teaspoons (tsp) (21 tsp for 6-11 years and 25 tsp for 12-18 years) accounting for 17% of total energy intake for both groups. The percent variance explained in BMI Z scores was 3.9% in children 6 to 11 years, with AS contributing only 0.03%. In children 12 to 18 years, the percent variance explained in the BMI Z scores was 6.5% with 0.18% coming from AS. No significant associations were observed between intake of AS and weight or adiposity measures. Consumption of AS did not contribute significantly to BMI Z scores in children 6 to 18 years. Longitudinal studies using more robust and precise measures of dietary intake are needed to further investigate the role of AS and weight in children. PMID- 21636012 TI - Nutritional supplementation of hop rho iso-alpha acids, berberine, vitamin D3, and vitamin K1 produces a favorable bone biomarker profile supporting healthy bone metabolism in postmenopausal women with metabolic syndrome. AB - Metabolic syndrome poses additional risk for postmenopausal women who are already at risk for osteoporosis. We hypothesized that a nutritional supplement containing anti-inflammatory phytochemicals and essential bone nutrients would produce a favorable bone biomarker profile in postmenopausal women with metabolic syndrome. In this 14-week, randomized trial, 51 women were instructed to consume a modified Mediterranean-style, low-glycemic-load diet and to engage in aerobic exercise. Those in the intervention arm (n = 25) additionally received 200 mg hop rho iso-alpha acids, 100 mg berberine sulfate trihydrate, 500 IU vitamin D3, and 500 MUg vitamin K1 twice daily. Forty-five women completed the study. Baseline nutrient intake did not differ between arms. Compared with baseline, the intervention arm exhibited an approximate 25% mean decrease (P < .001) in serum osteocalcin (indicative of bone turnover), whereas the placebo arm exhibited a 21% increase (P = .003). Serum 25-hydroxyvitamin D increased 23% (P = .001) in the intervention arm and decreased 12% (P = .03) in the placebo arm. The between arm differences for osteocalcin and 25-hydroxyvitamin D were statistically significant. Serum insulin-like growth factor I was statistically increased in both arms, but the between-arm differences were not statistically significant. Subanalysis showed that among those in the highest tertile of baseline insulin like growth factor I, the intervention arm exhibited a significant increase in amino-terminal propeptide of type I collagen, whereas the placebo arm showed a significant decrease at 14 weeks. Treatment with rho iso-alpha acids, berberine, vitamin D3, and vitamin K1 produced a more favorable bone biomarker profile indicative of healthy bone metabolism in postmenopausal women with metabolic syndrome. PMID- 21636013 TI - Low intake of vitamin B6 is associated with irritable bowel syndrome symptoms. AB - Most subjects with irritable bowel syndrome (IBS) experience an association between symptoms and food consumption. Although dietary intake has been the focus of previous research, attention to specific nutrients has been rare. We hypothesized that there is an association between the severity of IBS symptoms and the intake of specific food groups and specific nutrients. In this cross sectional study, 17 human subjects with IBS, as defined according to the Rome II criteria, were recruited. IBS symptoms were recorded on diary cards every evening for 7 days, and an IBS sum score was calculated (range, 0-15). Intake of food was assessed from a food diary kept by the subjects in the same period. Associations between IBS sum score and dietary intake were explored. The daily IBS sum score was 6.43 (range, 3.86- 9.09). Intake of vitamin B6 was the only component of the diet that was significantly associated with the IBS sum score. The median daily intake of vitamin B6 was 0.9 mg/day (range, 0.6-1.5), the recommended daily intake for men and women is 1.6 mg/day or more and 1.2 mg/day or more, respectively. A high symptom score was associated with low vitamin B6 intake (adjusted R2 = 0.583; beta = -4.431; 95% confidence interval, -6.386 to -2.476; P = 0.0002). A significant inverse association between intake of vitamin B6 and severity of IBS symptoms might have clinical implications. PMID- 21636014 TI - Obesity with a body mass index under 30 does not significantly impair the immune response in young adults. AB - Obesity accompanies various metabolic and immunologic changes. Evidence from epidemiological, animal, and human studies has linked obesity to impaired immunity. However, human studies that have investigated the immunocompetence of the obese are still limited. We studied the immune and inflammatory responses of obese (body mass index [BMI], 28.3 +/- 0.5 kg/m2; n = 30) and normal-weight (BMI, 21.2 +/- 0.3 kg/m2; n = 15) young adults to test the hypothesis that obesity is associated with an impaired immune function and dysregulated inflammatory response. Serum levels of adipokines and subpopulations of immune cells were examined. In vitro proliferative response of whole blood lymphocytes, the production of inflammatory cytokines (interleukin [IL] 1beta, IL-6, and tumor necrosis factor alpha) and T helper 1/T helper 2 cytokines (IL-2, interferon gamma, IL-4, and IL-10) were determined. Serum leptin levels were significantly higher (P < .001) and CD8+ T-cell subpopulation was significantly lower (P = .044) in the obese than normal-weight subjects. There was no difference in the proliferative response of whole-blood lymphocytes to T-cell mitogens between 2 groups. Phytohemagglutinin-stimulated peripheral blood mononuclear cells from the obese group produced significantly higher levels of IL-2 (P = .002) and tended to produce higher levels of IL-4 (P = .053) than those from the normal-weight group. No significant differences in the production of inflammatory cytokines by either whole-blood lymphocytes or peripheral blood mononuclear cells stimulated with lipopolysaccharide were observed between the obese and normal-weight subjects. These results suggest that obesity with a BMI less than 30 does not significantly impair immune function in young adults. However, further research is needed to investigate the clinical significance of a lower CD8+ T-cell population associated with obesity. PMID- 21636015 TI - Preliminary evidence that glucose ingestion facilitates prospective memory performance. AB - Previous research has found that the ingestion of glucose boosts task performance in the memory domain (including tasks tapping episodic, semantic, and working memory). The present pilot study tested the hypothesis that glucose ingestion would enhance performance on a test of prospective memory. In a between-subjects design, 56 adults ranging from 17 to 80 years of age performed a computerized prospective memory task and an attention (filler) task after 25 g of glucose or a sweetness-matched placebo. Blood glucose measurements were also taken to assess the impact of individual differences on glucose regulation. After the drink containing glucose, cognitive facilitation was observed on the prospective memory task after excluding subjects with impaired fasting glucose level. Specifically, subjects receiving glucose were 19% more accurate than subjects receiving a placebo, a trend that was marginally nonsignificant, F1,41 = 3.4, P = .07, but that had a medium effect size, d = 0.58. Subjects receiving glucose were also significantly faster on the prospective memory task, F1,35 = 4.8, P < .05, d = 0.6. In addition, elevated baseline blood glucose (indicative of poor glucose regulation) was associated with slower prospective memory responding, F1,35 = 4.4, P < .05, d = 0.57. These data add to the growing body of evidence suggesting that both memory and executive functioning can benefit from the increased provision of glucose to the brain. PMID- 21636016 TI - Weight loss increased serum adiponectin but decreased lipid levels in obese subjects whose body mass index was lower than 30 kg/m2. AB - We hypothesized that weight loss in obese subjects may affect adipokine levels, such as adiponectin and tumor necrosis factor (TNF) alpha. This study investigated the effects of an 8-week weight-control program on serum adiponectin, TNF-alpha, and blood lipid level profiles in obese subjects. Twenty obese subjects with a body mass index (BMI) higher than 25 kg/m2 were recruited for this weight loss program that used dietetic control and aerobic exercise training. A total of 3 obese men and 11 obese women (mean age, 40.3 +/- 10.8 years; BMI, 30.0 +/- 3.4 kg/m2) finished the program. Anthropometric and biochemical characteristics in subjects before and after the program were determined. The results showed that subjects' body weight, BMI, waist circumference, hip circumference, diastolic blood pressure, total cholesterol, and low-density lipoprotein cholesterol levels significantly (P < .05) decreased during the program. Further analysis showed a negative correlation between delta adiponectin and delta TNF-alpha, triacylglycerol, and systolic blood pressure in obese subjects. Subgroup analysis showed that obese subjects whose original BMI was less than 30 kg/m2 had significantly increased serum adiponectin levels, and more than 3% weight reduction markedly improved blood lipids and body fat profiles during the program. Our findings suggest that weight reduction through an 8-week weight loss program may have anti-inflammatory and antiatherogenic effects via increased serum adiponectin levels and improvements in blood lipid profiles and systolic blood pressure. PMID- 21636017 TI - Fermented garlic protects diabetic, obese mice when fed a high-fat diet by antioxidant effects. AB - This study examined the bioactivity of yeast (Saccharomyces cerevisiae)-fermented aged black garlic (FBG) on obese mice supplied a high-fat diet (HFD) and its in vitro antioxidant activity. Aged black garlic (BG) exhibits potent antioxidative effects and has been subjected to extensive research. In addition, the bioactivity of some natural products is increased by fermentation. In a preliminary test, this study found that the antioxidant activity of FBG is stronger than that of BG. Therefore, it was hypothesized that the bioactivity of BG would be increased by yeast fermentation and would be a good candidate as a nutraceutical product for improving the oxidative defense systems in older patients or patients affected by various oxidative stresses, for example, diabetes and diabetic complications. To test this hypothesis, the bioactivities of FBG in diabetic and obese mice as well as the antioxidant activity in vitro were examined. After 91 days of continuous HFD supply, the mice showed marked obesity, hyperglycemia, hyperlipemia, and liver and kidney damages. Black garlic and all 3 different doses of FBG showed favorable hepatoprotective, nephroprotective, hypolipidemic, and antiobesity effects compared with the HFD control, but no hypoglycemic effects. In particular, more favorable bioactivity against all 4 HFD-induced diabetic complications was detected in the FBG-treated groups compared with the group given equivalent doses of BG. These findings suggest that the bioactivities of BG can be improved by yeast fermentation. PMID- 21636018 TI - Consumption of legumes improves certain bone markers in ovariectomized rats. AB - Soybeans are known to protect against osteoporosis, but other legumes frequently consumed in Asia have not been studied to learn if they have a similar protective effect. This study investigated the hypothesis that consumption of soybean, mung bean, cowpea, and adzuki bean has beneficial effects on bone biomarkers in ovariectomized rats. Female Sprague-Dawley rats were either sham operated (sham; n = 7) or surgically ovariectomized and then fed a regular AIN-93M diet (OVX; n = 7) or AIN-93M containing soybean (n = 7), mung bean (n = 7), cowpea (n = 7), or adzuki beans (n = 7) for 10 weeks. No bean consumption significantly altered the body, subcutaneous fat, or uterus weight; however, consumption significantly increased the serum calcium/phosphorous ratio and decreased urinary calcium excretion compared with those of the OVX group. Serum concentration of 17beta estradiol was significantly lower in the OVX group compared with that of the sham group and was lowest in the group fed OVX diet containing soybean. Serum osteocalcin concentration was significantly higher in all OVX rats given a diet with beans compared with the same diet without, but urinary deoxypyridinoline excretion was lowest in the group fed OVX diet containing cowpea. There were no significant differences in bone mineral density or bone mineral content of the right femur, tibia, or lumbar spine or in the trabecular bone volume of the tibia among the diet groups. In conclusion, the consumption of soybean, mung bean, cowpea, and adzuki bean in OVX rats improved osteocalcin, but only those fed cowpea showed decreased bone resorption biomarker, suggesting that cowpea may have the most protective effect on bone in OVX rats. PMID- 21636019 TI - Rapid homeostatic response of H4IIE cells to diethylenetriaminepentaacetic acid is not due to changes in the amount or localization of ZnT-1 protein. AB - We have demonstrated that reducing zinc availability with the extracellular chelator diethylenetriaminepentaacetic acid (DTPA) causes rapid inhibition of cellular zinc efflux in H4IIE hepatoma cells but increases zinc efflux in primary hepatocytes. Similar differences were also observed between the rat anterior pituitary cell line GH3 and primary anterior pituitary cells. We hypothesized that the difference between the transformed and primary cells is due to differential regulation of ZnT-1 or SLC-30A-1 because this is the only zinc efflux transporter localized to the plasma membrane. The effects of DTPA (50 MUM) and zinc (100 MUM) treatment on messenger RNA (mRNA) and protein expression and protein localization of ZnT-1 were studied in H4IIE cells and primary hepatocytes. Although zinc tended to increase ZnT-1 mRNA in H4IIE cells, DTPA had no effect on ZnT-1 mRNA and protein expression in either hepatoma cells or hepatocytes. Although ZnT-1 is thought to be localized on the plasma membrane, this localization was not seen in these liver cells where ZnT-1 was distributed throughout the cytoplasm. Vesicular localization of ZnT-1 appeared to increase with zinc treatment. Total zinc content was reduced by DTPA in H4IIE cells. Diethylenetriaminepentaacetic acid also reduced metallothionein 1 mRNA, reflecting this reduction in cellular zinc. We conclude that the rapid homeostatic response of cells to altered zinc availability must be attributed to a transporter other than ZnT-1 or to changes in the activity of ZnT-1 by a novel mechanism. PMID- 21636020 TI - Universal total wrist arthroplasty: midterm follow-up study. AB - PURPOSE: We reviewed 21 consecutive patients who underwent a total wrist arthroplasty as a primary procedure between October 2001 and February 2007. The purposes of the present study were to communicate our midterm results and to compare them with previously published series. METHODS: We evaluated all patients clinically and radiologically. We used the Patient-Related Wrist Evaluation a primary outcome measure. The mean follow-up was 5.5 years (range, 3-8 years). A total of 14 patients had rheumatoid arthritis, including 1 with juvenile arthritis, and 1 each had psoriatic arthritis, systemic lupus erythematosus, and undifferentiated spondyloarthropathy. Of the remaining 4 patients, 2 had grade IV Kienbock disease, 1 had degenerative arthrosis, and 1 had chondrocalcinosis. RESULTS: Postoperative Patient-Related Wrist Evaluation scores averaged 24 points (SD, 21 pints) out of 100 (worst score). When the patients were specifically asked about pain and function of the arthroplasty, 20 claimed to be satisfied or very satisfied with the procedure. Two early and 3 late complications occurred. One patient had a wound hematoma and another had a superficial wound infection, both of which resolved with no further complications during the immediate postoperative period. In 2 patients, there was some osteolysis around the screw inserted into the medullary canal of the index metacarpal, but not in the trapezoid bone. One patient had a slight loosening of the distal component with subsidence on the ulnar side of the carpus. There have been no dislocations or surgical revisions of the components. CONCLUSIONS: Based on our study, a total wrist arthroplasty should be considered as a good alternative to arthrodesis for patients who wish to preserve some degree of mobility of the wrist. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic IV. PMID- 21636021 TI - Comparison of a new multifilament stainless steel suture with frequently used sutures for flexor tendon repair. AB - PURPOSE: To investigate the mechanical properties of some common suture materials currently in use and compare them with a new multifilament stainless steel suture. METHODS: We investigated the mechanical properties of 3-0 and 4-0 Fiberwire, 3-0 Supramid, 3-0 Ethibond, and a new 3-0 and 4-0 multifilament stainless steel suture. All suture material was tested in a knotted configuration and all but the Supramid was tested in an unknotted configuration. We measured the load, elongation at failure, and stiffness during both tests. RESULTS: The 4 0 multifilament stainless steel showed the least elongation, whereas the 3-0 multifilament stainless steel withstood the highest load of any material in both the knotted and unknotted tests. There was no difference in stiffness between the 3-0 and 4-0 multifilament stainless steel when untied; however, the 3-0 multifilament stainless steel was stiffer when tied. Soaking in a saline solution had no significant effect on the ultimate load, elongation at failure, or stiffness of any of the sutures. The 3-0 Fiberwire and 3-0 Ethibond required at least 5 throws to resist untying. CONCLUSIONS: Multifilament stainless steel exhibited promising mechanical advantages over the other sutures tested. More research is needed to determine how this material will affect the clinical outcomes of primary flexor tendon repair. CLINICAL RELEVANCE: With a secure attachment to the tendon, the multifilament stainless steel's lower elongation and better knot-holding ability may result in a higher force to produce a 2-mm gap and a higher ultimate tensile strength in a tendon repair. PMID- 21636022 TI - Stenosing synovitis of the extensor pollicis longus tendon. AB - There are only a few published cases of extensor pollicis longus (EPL) tenosynovitis in patients without rheumatoid arthritis. Even less common are cases of stenosing tenosynovitis of the EPL associated with triggering. This article presents 2 cases of EPL stenosing tenosynovitis with triggering of the thumb in the area of Lister's tubercle and addresses how to treat them. PMID- 21636023 TI - Predictors of scar pain after open carpal tunnel release. AB - PURPOSE: To identify the predictors of scar pain after open carpal tunnel release (CTR). METHODS: We enrolled 83 patients with idiopathic carpal tunnel syndrome treated by open CTR. All patients completed the Brigham and Women's (Boston) carpal tunnel questionnaire (BCTQ) preoperatively. We assessed levels of depression preoperatively using the Center for the Epidemiological Study of Depression (CES-D) scale, and pain anxiety using the Pain Anxiety Symptoms Scale. At 3 months after surgery, patients were asked to self-assess treatment satisfaction and scar pain using a 10-point ordinal scale and to complete the BCTQ. RESULTS: The mean BCTQ-symptom (BCTQ-S) score decreased significantly from 2.7 +/- 1.1 preoperatively to 1.6 +/- 1.0 at 3 months postoperatively, and mean BCTQ-function score decreased significantly from 2.4 +/- 1.1 to 1.4 +/- 1.0. Overall, scar pain intensity at 3 months postoperatively ranged from 0 to 8 (mean, 2.4 +/- 2.2), and overall satisfaction ranged from 2 to 10 (mean, 7.6 +/- 2.6). The intensity of the scar pain was significantly correlated with the CES-D scale and BCTQ-S. Multivariable regression analysis showed that depression, assessed using the CES-D scale, and postoperative symptoms, assessed using the BCTQ-S, predicted scar pain intensity, which accounted for 38% of scar pain intensity variance. CONCLUSIONS: Depression score and postoperative symptoms predicted scar pain intensity after open CTR. However, the most important contributor to scar pain intensity variance remains unidentified. TYPE OF STUDY/LEVEL OF EVIDENCE: Prognostic I. PMID- 21636024 TI - Treatment of bony boutonniere deformity with a loop wire. AB - Boutonniere deformity associated with a dorsal avulsion fracture of the central slip overlying the proximal interphalangeal joint results in loss of extension of the joint and hyperextension of the distal interphalangeal joint. This article reports a surgical technique for treatment of the injury in 21 digits, which involves application of loop stainless steel wire. We also present the long-term results using the technique on the digits. Loop wire fixation is a successful surgical technique for the treatment of displaced central slip avulsion fracture. PMID- 21636025 TI - Microsurgery of the upper extremity. AB - In the past 50 years, hand surgeons have made considerable contributions to microsurgery. The unique demands of complex upper extremity care have driven many of the technical and scientific advances of this discipline, including functional muscle transfers, nerve transfers, and composite tissue allotransplantation. The purpose of this article was to review the current applications of microsurgery to the upper extremity. PMID- 21636026 TI - Replantation surgery. AB - The current concepts of replantation surgery, a procedure that has been practiced for half a century, can be discussed in terms of patients' demands and expectations, present indications for the procedure, available evidence that influences decision making, and technical refinements practiced to produce better outcomes. PMID- 21636027 TI - Anatomical study of the A1 pulley: length and location by means of cutaneous landmarks on the palmar surface. PMID- 21636028 TI - Wire loop technique to retrieve flexor tendon. PMID- 21636029 TI - A traumatic rupture of valsalva sinus with dissection into the interventricular septum. PMID- 21636030 TI - Triad of metabolic syndrome, chronic kidney disease, and coronary heart disease with a focus on microalbuminuria death by overeating. AB - Coronary heart disease remains a major cause of morbidity and mortality in the United States, and its incidence is rising worldwide. Because atherosclerosis is a chronic process, and it is often associated with certain lifestyle and risk factors such as hypertension, dyslipidemia, and insulin resistance, much emphasis is being placed on lifestyle modification and control of risk factors. It is being recognized that some lifestyle patterns such as overeating result in metabolic syndrome, which may play a role in the development of chronic kidney disease and coronary heart disease. Here, we focus on an important relationship between these 3 conditions, and we provide evidence that microalbuminuria develops in many patients with metabolic syndrome, may be an important correlate of chronic kidney disease and coronary heart disease, and may represent an important prognostic marker. Although the pathogenesis of microalbuminuria in metabolic syndrome is not clear, we suggest that microalbuminuria, chronic kidney disease, and coronary heart disease share common pathways involving inflammation and oxidative stress. We also discuss that a healthy lifestyle is essential for preventing and treating chronic kidney disease and coronary heart disease seen in patients with metabolic syndrome. PMID- 21636031 TI - Effect of switching antithrombin agents for primary angioplasty in acute myocardial infarction: the HORIZONS-SWITCH analysis. AB - OBJECTIVES: We investigated the outcomes of switching to bivalirudin after initial administration of heparin in patients with acute ST-segment elevation myocardial infarction undergoing primary percutaneous coronary intervention. BACKGROUND: Unfractionated heparin (UFH) is frequently administered early in ST segment elevation myocardial infarction. Whether the benefits of bivalirudin documented in the HORIZONS-AMI (Harmonizing Outcomes With Revascularization and Stents in Acute Myocardial Infarction) trial persist in patients previously administered UFH is unknown. METHODS: We analyzed the outcomes of the 2,357 patients from HORIZONS-AMI treated with UFH before enrollment according to their subsequent randomization to bivalirudin (switch group, n = 1,178) or UFH plus a glycoprotein IIb/IIIa inhibitor (control group, n = 1,179). RESULTS: At 30 days, major bleeding occurred in 7.6% of the switch group versus 12.3% of the control group (p = 0.0001). Switch patients had lower 30-day rates of cardiac mortality (1.6% vs. 2.9%, p = 0.04). At 2-year follow-up, switch patients experienced lower rates of major bleeding (8.4% vs. 13.0%, p = 0.0003), cardiac mortality (2.3% vs. 3.8%, p = 0.04), and reinfarction (4.0% vs. 7.1%, p = 0.0002). Two-year rates of definite/probable stent thrombosis were similar in switch and control patients (3.1% vs. 4.3%, p = 0.17). CONCLUSIONS: In ST-segment elevation myocardial infarction patients who receive early treatment with UFH, switching to bivalirudin before primary percutaneous coronary intervention results in reduced rates of major bleeding and improved early and late cardiac survival. PMID- 21636032 TI - Distinguishing arrhythmogenic right ventricular cardiomyopathy/dysplasia associated mutations from background genetic noise. AB - OBJECTIVES: The aims of this study were to determine the spectrum and prevalence of "background genetic noise" in the arrhythmogenic right ventricular cardiomyopathy/dysplasia (ARVC) genetic test and to determine genetic associations that can guide the interpretation of a positive test result. BACKGROUND: ARVC is a potentially lethal genetic cardiovascular disorder characterized by myocyte loss and fibrofatty tissue replacement of the right ventricle. Genetic variation among the ARVC susceptibility genes has not been systematically examined, and little is known about the background noise associated with the ARVC genetic test. METHODS: Using direct deoxyribonucleic acid sequencing, the coding exons/splice junctions of PKP2, DSP, DSG2, DSC2, and TMEM43 were genotyped for 93 probands diagnosed with ARVC from the Netherlands and 427 ostensibly healthy controls of various ethnicities. Eighty-two additional ARVC cases were obtained from published reports, and additional mutations were included from the ARVD/C Genetic Variants Database. RESULTS: The overall yield of mutations among ARVC cases was 58% versus 16% in controls. Radical mutations were hosted by 0.5% of control individuals versus 43% of ARVC cases, while 16% of controls hosted missense mutations versus a similar 21% of ARVC cases. Relative to controls, mutations in cases occurred more frequently in non-Caucasians, localized to the N-terminal regions of DSP and DSG2, and localized to highly conserved residues within PKP2 and DSG2. CONCLUSIONS: This study is the first to comprehensively evaluate genetic variation in healthy controls for the ARVC susceptibility genes. Radical mutations are high-probability ARVC-associated mutations, whereas rare missense mutations should be interpreted in the context of race and ethnicity, mutation location, and sequence conservation. PMID- 21636033 TI - Hearing the noise the challenges of human genome variation in genetic testing. PMID- 21636034 TI - Preventing overdiagnosis of implantable cardioverter-defibrillator lead fractures using device diagnostics. AB - OBJECTIVES: This study sought to use implantable cardioverter-defibrillator (ICD) diagnostics to discriminate ICD lead fractures from normally functioning leads with high impedance and from connection problems between the lead and header. BACKGROUND: ICD diagnostics facilitate identification of fractures, but there are no accepted criteria for discriminating fractures from other causes of high impedance and/or nonphysiological "noise" oversensing. METHODS: We analyzed a development set of 91 leads to construct a stepwise algorithm based on ICD diagnostics. It included 40 fractures, 30 connection problems, and 21 functioning leads that triggered high-impedance alerts. Then we applied this algorithm to an independent test set of 100 leads: 70 fractures and 30 intact leads with connection problems that were misdiagnosed clinically as fractures. In the algorithm, either extremely high maximum impedance or noise oversensing with a normal impedance trend indicated a fracture. A short interval from surgery to impedance rise or prolonged stable impedance after an abrupt rise indicated a connection problem. A gradual impedance increase or stable, high impedance indicated a functioning lead. RESULTS: In the test set, the algorithm correctly classified 100% of fractures (95% confidence interval [CI]: 95% to 100%) and 87% of connection problems that were misdiagnosed as fractures (95% CI: 70% to 95%). CONCLUSIONS: An algorithm using only ICD diagnostics identifies leads with oversensing or high impedance as fractures or connection problems with a high degree of accuracy. PMID- 21636035 TI - Low prevalence of risk markers in cases of sudden death due to Brugada syndrome relevance to risk stratification in Brugada syndrome. AB - OBJECTIVES: The objective of this study was to determine the prevalence of conventional risk factors in sudden arrhythmic death syndrome (SADS) probands with Brugada syndrome (BrS). BACKGROUND: Patients with BrS and previous aborted sudden cardiac death (SCD) are at high risk of recurrent events. Other universally accepted clinical features associated with higher risk include unheralded syncope and the presence of a spontaneous type 1 electrocardiogram (ECG). METHODS: We analyzed reported symptoms and reviewed ECGs from SADS probands with familial diagnoses of BrS, established by cardiological evaluation, including ECG, 2-dimensional echocardiography, Holter monitoring, exercise tolerance testing, and ajmaline provocation. These cases underwent familial evaluation between 2003 and 2010. RESULTS: A total of 49 consecutive families with a confirmed SADS death and a diagnosis of BrS were evaluated, comprising assessment of 202 family members in total. One family had 2 members with SADS, resulting in a total of 50 probands included. Mean age of death of probands was 29.1 +/- 10.6 years, with 41 males (82%) (p < 0.05). Antemortem ECGs were available for 5 SADS probands, 1 of which demonstrated a spontaneous type 1 pattern. In 45 probands, symptoms before death were reported reliably by family members. Of these, 9 (20%) had experienced at least 1 syncopal episode before the fatal event. Importantly, 68% of probands would not have fulfilled any current criteria for consideration of implantable cardioverter-defibrillator. CONCLUSIONS: The "low-risk" asymptomatic BrS group comprises the majority of SCD in this cohort. Current risk stratification would appear to be inadequate, and new markers of risk are vital. PMID- 21636036 TI - Clinical implications of midventricular obstruction in patients with hypertrophic cardiomyopathy. AB - OBJECTIVES: We investigated the prevalence, clinical characteristics, and prognosis of hypertrophic cardiomyopathy (HCM) patients with midventricular obstruction (MVO). BACKGROUND: Previous descriptions of patients with MVO have been confined to case reports or small patient series, and this subgroup of HCM patients has therefore remained underrecognized. METHODS: The study population included 490 HCM patients. Left ventricular MVO was diagnosed when the peak midcavitary gradient was estimated to be >=30 mm Hg. RESULTS: MVO was identified in 46 patients (9.4%). Patients with MVO were more likely to be symptomatic than those without. MVO was found to be an independent determinant of HCM-related death in multivariate models (hazard ratio [HR]: 2.23, p = 0.016), and this trend was especially pronounced for the combined endpoint of sudden death and potentially lethal arrhythmic events (HR: 3.19, p < 0.001). Apical aneurysm formation was identified in 28.3% of patients with MVO and strongly predicted HCM related death (HR: 3.47, p = 0.008) and the combined endpoint of sudden death and potentially lethal arrhythmic events (HR: 5.08, p < 0.001). In addition, MVO without apical aneurysm was also identified as an independent determinant of the combined endpoint of sudden death and potentially lethal arrhythmic events (HR: 2.43, p = 0.045). CONCLUSIONS: This analysis identified MVO as an independent predictor of adverse outcomes, especially the combined endpoint of sudden death and potentially lethal arrhythmic events. Our results suggest that longer periods of exposure to MVO might lead to unfavorable consequences. They also support the principle that the presence of MVO in patients with HCM has important pathophysiological implications. PMID- 21636038 TI - The importance of functional status in the management of peripheral arterial disease. PMID- 21636037 TI - Greater sedentary hours and slower walking speed outside the home predict faster declines in functioning and adverse calf muscle changes in peripheral arterial disease. AB - OBJECTIVES: In participants with peripheral arterial disease (PAD), we determined whether more sedentary behavior and slower outdoor walking speed were associated with faster functional decline and more adverse changes in calf muscle characteristics over time. BACKGROUND: Modifiable behaviors associated with faster functional decline in lower-extremity PAD are understudied. METHODS: Participants were 384 men and women with an ankle brachial index <0.90 followed for a median of 47 months. At baseline, participants reported the number of hours they spent sitting per day and their walking speeds outside their homes. Participants underwent baseline and annual measures of objective functional performance. Calf muscle characteristics were measured with computed tomography at baseline and every 2 years subsequently. Analyses were adjusted for age, sex, race, comorbidities, ankle brachial index, and other confounders. RESULTS: Slower walking speed outside the home was associated with faster annual decline in calf muscle density (brisk/striding pace -0.32 g/cm(3), average pace -0.46 g/cm(3), casual strolling -1.03 g/cm(3), no walking at all -1.43 g/cm(3), p trend <0.001). Greater hours sitting per day were associated with faster decline in 6-min walk (<4 h: -35.8 feet/year; 4 to <7 h: -41.1 feet/year; 8 to <11 h: -68.7 feet; >=12 h: -78.0 feet; p trend = 0.008). Similar associations were observed for greater hours sitting per day and faster declines in fast-paced (p trend = 0.018) and usual-paced (p trend < 0.001) 4-m walking velocity. CONCLUSIONS: Greater sedentary hours per day and slower outdoor walking speed are modifiable behaviors that are associated with faster functional decline and greater decline in calf muscle density, respectively, in patients with PAD. PMID- 21636039 TI - Transfer of adolescents with congenital heart disease from pediatric cardiology to adult health care: an analysis of transfer destinations. AB - OBJECTIVES: The transfer of adolescents with congenital heart disease from pediatric to adult care was examined. The aims were to investigate where these adolescents received adult-centered care, to determine the proportion of patients with no follow-up and with no appropriate follow-up after leaving pediatric cardiology, and to explore the determinants of no follow-up and no appropriate follow-up. BACKGROUND: Even after successful treatment, many patients require lifelong cardiac surveillance by specialized practitioners. Although guidelines describe the most appropriate level of follow-up, this is not always implemented in practice. METHODS: A descriptive, observational study was performed, including 794 patients with congenital heart disease examined and/or treated at a tertiary care center. RESULTS: Overall, 58 of the 794 patients included (7.3%) were not in follow-up. Cessation of follow-up was found in 2 of 74 patients with complex (2.7%), 31 of 448 patients with moderate (6.9%), and 25 of 272 patients with simple (9.2%) heart defects. Moreover, 684 patients (86.1%) remained in specialized follow-up. According to international guidelines, 81 patients (10.2%) did not receive the minimal level of cardiac care. Multivariable logistic regression revealed that male sex and no prior heart surgery were associated with no follow-up. Male sex, no prior heart surgery, and greater complexity of congenital heart disease were associated with no appropriate level of cardiac follow-up. CONCLUSIONS: The proportion of patients in this study lost to follow up was substantially lower than in other Western countries. Because only patient related factors were examined with respect to loss to follow-up, further examination of patient-related, hospital-related, and healthcare-related determinants of lack of follow-up is needed. PMID- 21636040 TI - Surgically corrected right-sided aortic arch. PMID- 21636041 TI - President's page: clouds, science, and education. PMID- 21636042 TI - Methodological issues in pharmacotherapy research in older adults. AB - BACKGROUND: The shifting of demographics to an aging society with multiple chronic illnesses and increased heterogeneity has created an undeniable imperative regarding the knowledge about pharmacotherapy in older patients, especially because older adults have a higher rate of medication use compared with the general population. More research on pharmacotherapy in older adults is needed to maximize not only the appropriate use of medications but also the benefits of available treatments in this historically underrepresented group. Investigators face many challenges that may greatly affect the outcomes of research on pharmacotherapy in older adults. OBJECTIVE: This commentary discusses the common challenges of research on pharmacotherapy in older adults and provides strategies to overcome such challenges. METHODS: The following databases were searched for examples of concepts (dates: 1976-September 14, 2010; key terms: research [in the title], aged, elderly, older adult, and geriatric): PubMed, Cumulative Index to Nursing and Allied Health Literature Database (CINAHL), Family & Societal Studies Worldwide, Women's Studies International, Academic Search Premier, Health Source: Academic/Nursing Edition, ISI Web of Knowledge, International Pharmaceutical Abstracts, PsycInfo, and Social Work Abstracts. RESULTS: There are several methodological challenges related to study design, including the measurement of outcomes and the analytical plan; study implementation, such as the recruitment of sites and individuals; the informed consent process; retention; and other practical considerations. A crucial area to consider is the measurement of medication adherence, both as an independent factor and as a confounder. Many challenges of research in older patients reflect the complexities of the aging process, including psychosocial consequences, and the resultant effects on study participation and outcomes. CONCLUSION: It is hoped that the recommendations provided will help researchers and clinicians overcome the methodological challenges of conducting research on pharmacotherapy in older adults, thereby fostering more studies that critically evaluate pharmacotherapy and medication-management services and maximizing treatment outcomes in older adults. PMID- 21636044 TI - Re: "outsourcing to teleradiology companies: bad for radiology, bad for radiologists". PMID- 21636046 TI - Without a strong team, we cannot advance. PMID- 21636047 TI - The seed corn. PMID- 21636048 TI - Reading portable radiographs: a call for honesty. PMID- 21636049 TI - Academic job market. PMID- 21636050 TI - The innovator's solution. PMID- 21636051 TI - ACR Appropriateness Criteria((r)) on suspected lower extremity deep vein thrombosis. AB - Lower extremity deep vein thrombosis (DVT) is a common clinical concern, with an incidence that increases with advanced age. DVT typically begins below the knee but may extend proximally and result in pulmonary embolism. Pulmonary embolism can occur in 50% to 60% of patients with untreated DVT and can be fatal. Although clinical examination and plasma d-dimer blood evaluation can often predict the presence of DVT, imaging remains critical for the diagnostic confirmation and treatment planning of DVT. Patients with above-the-knee or proximal DVT have a high risk for pulmonary embolism and are recommended to receive anticoagulation therapy. On the other hand, patients with below-the-knee or distal DVT rarely experience pulmonary embolism, and anticoagulation therapy in these patients remains controversial. However, one sixth of patients with distal DVT may experience extension of their thrombus above the knee and therefore are recommended to undergo serial imaging assessment at 1 week to exclude proximal DVT extension if anticoagulation therapy is not initiated. Ultrasound is the preferred imaging method for evaluation of patients with newly suspected lower extremity DVT. Magnetic resonance and CT venography can be especially helpful for the evaluation of suspected DVT in the pelvis and thigh. Contrast x-ray venography, the historic gold standard for DVT assessment, is now less commonly performed and primarily reserved for patients with more complex presentations such as those with suspected recurrent acute DVT. PMID- 21636052 TI - ACR-SNM Task Force on Nuclear Medicine Training: report of the task force. AB - The expansion of knowledge and technological advances in nuclear medicine and radiology require physicians to have more expertise in functional and anatomic imaging. The convergence of these two specialties into the new discipline of molecular imaging has also begun to place demands on residency training programs for additional instruction in physiology and molecular biology. These changes have unmasked weaknesses in current nuclear medicine and radiology training programs. Adding to the impetus for change are the attendant realities of the job market and uncertain employment prospects for physicians trained in nuclear medicine but not also trained in diagnostic radiology. With this background, the ACR and the Society of Nuclear Medicine convened the Task Force on Nuclear Medicine Training to define the issues and develop recommendations for resident training. PMID- 21636053 TI - Radiology benefit managers: cost saving or cost shifting? AB - PURPOSE: Radiology benefit managers (RBMs) are widely used by private payers to manage the utilization of imaging services through prior authorization, and they have been proposed for use in the Medicare program. The authors created a framework for evaluating the impact of key parameters on the ability of RBMs to lower costs and used decision-analytic modeling to simulate the net impact of RBMs on health care costs under uncertainty from a societal perspective. METHODS: The authors' model of a "typical" RBM's prior authorization process used base case values for each parameter (utilization rate and costs for MR, CT, and PET imaging; physician and staff time spent in complying with RBM requirements; approval and denial rates; and RBM fees to insurers) drawn from published data and the experience of a large, academic institution. Different values were tested in the sensitivity analysis to account for uncertainty in the parameter estimates. A hypothetical 100,000-member private health plan with an imaging utilization rate of 135 per 1,000 members per year was assumed. RESULTS: Under the authors' base-case scenario, in which RBMs have no net impact on costs, they estimated that 28% ($182,066/$640,263) of the projected RBM-related savings are shifted to providers. RBMs were cost saving in 45% of simulations, and 95% of simulations fell between a cost decrease of $397,880 and a cost increase of $341,991. The probability of an initial approval by the RBM, the RBM's fee, and the imaging utilization rate and associated charges had the largest influence on the results. CONCLUSIONS: The authors' models shows that RBMs shift significant costs to physicians and that their net impact on societal costs depends on parameters for which supporting data are incomplete. PMID- 21636054 TI - Informing patients about risks and benefits of radiology examinations: a review article. AB - Communicating the risks, benefits, and alternatives to a planned medical intervention is integral to high-quality patient care. When effective, such communication promotes patient autonomy, alleviates unfounded patient apprehension, and mitigates medicolegal liability. The topic of medical radiation adds to the usual challenges of effective medical communication some special challenges of its own. Among these is a lack of understanding by the general population and health professionals of the benefits and risks of medical radiation, which is compounded by unfamiliar terminology and units of measure. This is further complicated by the fact that many patients have poor comprehension of risk data in general. In this article, the authors present a case, review the ethical basis and legal history of informed consent, and explore the current initiatives, available resources, and further opportunities related to this challenging topic. PMID- 21636055 TI - Cross-sectional examination interpretation discrepancies between on-call diagnostic radiology residents and subspecialty faculty radiologists: analysis by imaging modality and subspecialty. AB - PURPOSE: The aim of this study was to identify the rate of discrepancies between radiology residents and faculty radiologists at an academic hospital and to determine the distribution across subspecialties and modalities, specifically CT, MR, and ultrasound. METHODS: Consecutive CT, MR, and ultrasound preliminary interpretations rendered by on-call second-year through fourth-year radiology residents for 9 months on emergency department patients, inpatients, and urgent outpatients formed the study population. All preliminary interpretations were graded using a modified RADPEER((r)) system (scores 0 and 1 = concordance; score 2 = minor, clinically insignificant discordance; scores 3 and 4 = clinically significant discordance) by the subspecialty faculty members who rendered the final interpretation. RESULTS: There were 158 clinically significant discrepancies out of 21,482 preliminary interpretations, for a discrepancy rate of 0.7%. There was no statistically significant difference in rates across subspecialties or between adult and pediatric examinations (cardiothoracic, 1%; abdominal, 0.7%; neuroradiology, 0.6%; musculoskeletal, 0.7%; pediatrics, 0.8%). MR and CT interpretations had significantly higher rates than ultrasound (MR, 1.4%; CT, 0.9%; ultrasound, 0.2%; P < .001). Within neuroradiology, there was a significantly higher rate for MR than CT (1.5% vs 0.6%, P < .01), and within abdominal radiology, there was a significantly higher rate for CT than ultrasound (1.1% vs 0.2%, P < .01). CONCLUSIONS: Discrepancy rates in this study were less than or comparable with those reported previously and within or lower than rates for practicing radiologists. Discrepancy rates varied among subspecialties and modalities, suggesting the use of a RADPEER system with attention to modality and subspecialty as a methodology for identifying areas for targeted resident education. PMID- 21636056 TI - Comparison of discrepancy rates in resident and faculty interpretations of on call PE CT and V/Q scans: is one study more reliable during off hours? AB - PURPOSE: There has been no comparison of concordance rates for residents' and faculty members' interpretations of pulmonary embolism (PE) CT and ventilation/perfusion (V/Q) studies. If significantly different rates of agreement are demonstrable, this could influence which test is ordered during off hours. The purpose of this study was to evaluate how the performance of residents compared with that of faculty members in interpreting these two modalities. METHODS: Interobserver agreement between preliminary resident reports and final attending radiologist interpretations was calculated retrospectively for PE CT studies (n = 1,179) and V/Q scans (n = 331) interpreted by on-call residents from April 2007 to October 2008. Discordant cases were reviewed for clinical significance and outcomes at 3 months. Interobserver variability was also evaluated relative to residents' training levels and faculty members' years of experience. RESULTS: Interobserver agreement between faculty members and residents was substantial, at 95% for PE CT (kappa = 0.77) and 77.9% for V/Q scans (kappa = 0.67). Although changes in interpretation were significantly more common with V/Q scans (22.1% vs 5%; chi(2) < .0001), the rates of clinically significant discrepancy did not differ significantly between the modalities (2.1% for V/Q vs 1.2% for PE CT; chi(2)P = .20). The overall discrepancy rate and the rate of clinically significant discrepancy did not correlate with residents' training levels or faculty members' years of experience. CONCLUSIONS: Concordance rates for residents' interpretations of V/Q scans and PE CT studies were high, and discrepancies resulting in changes in patient management were rare for both modalities. Residents' preliminary interpretations of both modalities are reliable and safe for making initial patient management decisions. PMID- 21636057 TI - The Clinical Research Center: a vital part of the ACR mission. AB - The ACR's mission statement identifies five pillars of excellence. One of its pillars is research. ACR is recognized by many as supporting one of the premier research endeavors sponsored by a professional medical society of which the ACR Clinical Research Center is the largest component. The center is comprised of four entities: ACRIN((r)), RTOG((r)), QRRO((r)), and ACR Image MetrixTM. The Clinical Research Center encompasses personnel with extensive clinical trial expertise, a state-of-the-art IT infrastructure, and an imaging and radiation oncology core laboratory. This research enterprise supports a global network of researchers in the conduct of medical imaging and radiation oncology clinical trials. This paper's focus is on the Clinical Research Center's value to the radiology and radiation oncology professions, to the practices engaged in the clinical research, and to our patients. PMID- 21636058 TI - The effect of imaging capacity on the imaging workup of patients with stroke: a preliminary case for a cross-country comparison. AB - PURPOSE: Medical imaging is a large and growing component of health care expenditures. To better understand some of the determinants of imaging ordering behavior, the authors analyzed the effect of differential capacity on the imaging workup of patients with acute nonhemorrhagic stroke. METHODS: All patients at a US teaching hospital and a two-campus Canadian teaching hospital between 2001 and 2005 discharged with diagnoses of acute nonhemorrhagic stroke were identified. Billing data were linked with clinical information systems to identify all imaging studies performed, comorbidities, and patient disposition. RESULTS: Nine hundred eighteen patients at the US hospital and 1,759 patients at the Canadian hospital were included. Patients were similar in age and distribution of comorbid illnesses. The rate of MRI scans at the US hospital was more than twice that at either of the Canadian hospitals (95.75 scans per 100 patients vs 41.39 scans per 100 patients). The length of stay was significantly shorter and the inpatient mortality rate significantly lower at the US hospital compared with the Canadian hospital. A multivariate regression analysis demonstrated that only patient age and site (US vs Canada) were significant predictors of MRI use, controlling for patient gender, comorbidities, and use of anticoagulants. CONCLUSIONS: Scanning utilization varied at hospitals with differential access to scanning technologies. There was less frequent use of MRI scanning at hospitals with limited access to this modality. Patient and health system factors are important considerations when interpreting the mechanisms for this variation, its importance, and the potential relationship of imaging use with patient outcomes. PMID- 21636059 TI - Natural language processing: the basics (part 1). PMID- 21636060 TI - Clinical audits: the European Society of Radiology Perspective. PMID- 21636061 TI - The lean concept of waste in radiology. PMID- 21636062 TI - Automatic exposure control in CT: applications and limitations. PMID- 21636063 TI - The heart of radiology in Minnesota. PMID- 21636064 TI - Morris Simon. PMID- 21636065 TI - Cognitive consequences of motivational orientation: perceived similarity between objects. AB - Recent findings suggest that the unconscious activation of the motivational orientations of approach and avoidance is accompanied by the adoption of a more global and a more local processing style, respectively. A global processing style, in turn, is assumed to instigate a focus on similarities whereas a local processing style is assumed to instigate a focus on differences. Integrating these two ideas, the present research examines the hypothesis that participants under approach perceive objects as more similar to each other than participants under avoidance. To test this assumption, we induced the two motivational orientations and elicited judgments of similarities (Experiments 1 and 2) and differences (Experiment 2) for pairs of pictures. Results confirmed the hypothesis. We propose that the relative attunement to similarities/differences under approach/avoidance is functional because it allows for a flexible conceptualization of the environment/an ability to discern slight deviations from what is expected. PMID- 21636066 TI - Mutations in FYCO1 cause autosomal-recessive congenital cataracts. AB - Congenital cataracts (CCs), responsible for about one-third of blindness in infants, are a major cause of vision loss in children worldwide. Autosomal recessive congenital cataracts (arCC) form a clinically diverse and genetically heterogeneous group of disorders of the crystalline lens. To identify the genetic cause of arCC in consanguineous Pakistani families, we performed genome-wide linkage analysis and fine mapping and identified linkage to 3p21-p22 with a summed LOD score of 33.42. Mutations in the gene encoding FYVE and coiled-coil domain containing 1 (FYCO1), a PI(3)P-binding protein family member that is associated with the exterior of autophagosomes and mediates microtubule plus-end directed vesicle transport, were identified in 12 Pakistani families and one Arab Israeli family in which arCC had previously been mapped to the overlapping CATC2 region. Nine different mutations were identified, including c.3755 delC (p.Ala1252AspfsX71), c.3858_3862dupGGAAT (p.Leu1288TrpfsX37), c.1045 C>T (p.Gln349X), c.2206C>T (p.Gln736X), c.2761C>T (p.Arg921X), c.2830C>T (p.Arg944X), c.3150+1 G>T, c.4127T>C (p.Leu1376Pro), and c.1546C>T (p.Gln516X). Fyco1 is expressed in the mouse embryonic and adult lens and peaks at P12d. Expressed mutant proteins p.Leu1288TrpfsX37 and p.Gln736X are truncated on immunoblots. Wild-type and p.L1376P FYCO1, the only missense mutant identified, migrate at the expected molecular mass. Both wild-type and p. Leu1376Pro FYCO1 proteins expressed in human lens epithelial cells partially colocalize to microtubules and are found adjacent to Golgi, but they primarily colocalize to autophagosomes. Thus, FYCO1 is involved in lens development and transparency in humans, and mutations in this gene are one of the most common causes of arCC in the Pakistani population. PMID- 21636067 TI - X-linked congenital hypertrichosis syndrome is associated with interchromosomal insertions mediated by a human-specific palindrome near SOX3. AB - X-linked congenital generalized hypertrichosis (CGH), an extremely rare condition characterized by universal overgrowth of terminal hair, was first mapped to chromosome Xq24-q27.1 in a Mexican family. However, the underlying genetic defect remains unknown. We ascertained a large Chinese family with an X-linked congenital hypertrichosis syndrome combining CGH, scoliosis, and spina bifida and mapped the disease locus to a 5.6 Mb critical region within the interval defined by the previously reported Mexican family. Through the combination of a high resolution copy-number variation (CNV) scan and targeted genomic sequencing, we identified an interchromosomal insertion at Xq27.1 of a 125,577 bp intragenic fragment of COL23A1 on 5q35.3, with one X breakpoint within and the other very close to a human-specific short palindromic sequence located 82 kb downstream of SOX3. In the Mexican family, we found an interchromosomal insertion at the same Xq27.1 site of a 300,036 bp genomic fragment on 4q31.2, encompassing PRMT10 and TMEM184C and involving parts of ARHGAP10 and EDNRA. Notably, both of the two X breakpoints were within the short palindrome. The two palindrome-mediated insertions fully segregate with the CGH phenotype in each of the families, and the CNV gains of the respective autosomal genomic segments are not present in the public database and were not found in 1274 control individuals. Analysis of control individuals revealed deletions ranging from 173 bp to 9104 bp at the site of the insertions with no phenotypic consequence. Taken together, our results strongly support the pathogenicity of the identified insertions and establish X linked congenital hypertrichosis syndrome as a genomic disorder. PMID- 21636068 TI - Use of in-cycle antimullerian hormone levels to predict cycle outcome. AB - OBJECTIVE: The goal of this work is to expand the usefulness of antimullerian hormone (AMH) in predicting in vitro fertilization cycle outcome by demonstrating that AMH concentration obtained in an ongoing treatment cycle predicts both oocyte number and pregnancy. STUDY DESIGN: Serum samples were obtained from 190 in vitro fertilization patients at onset of follicle-stimulating hormone stimulation. These were analyzed retrospectively during a single cycle in which clinicians were blinded to the results. Our major outcome measures were the number of oocytes obtained and ongoing pregnancy. RESULTS: Patients with an initial AMH concentration of >3 ng/mL were found to produce a mean of 19.8 oocytes and had an ongoing pregnancy rate of 60.3%. In contrast, those with AMH values of <=1 ng/mL yielded a mean of 6.2 oocytes and had an ongoing pregnancy rate of 23.4% (P < .0001 for both). CONCLUSION: Greater AMH serum concentration strongly predicts an increased number of oocytes and ongoing pregnancy (P <= .0001). PMID- 21636070 TI - Long-term complications associated with glaucoma drainage devices and Boston keratoprosthesis. AB - PURPOSE: To evaluate long-term complications related to glaucoma drainage devices in patients undergoing Boston type 1 keratoprosthesis surgery. DESIGN: Retrospective case series. METHODS: All patients who underwent Boston type 1 keratoprosthesis surgery at the University of California, Davis, between 2004 and 2010 were included. Preoperative and postoperative data were reviewed. Twenty five eyes with glaucoma drainage devices were highlighted. Visual acuity and postoperative complications were tracked at postoperative months 1, 3, 6, 9, and 12 and at annual intervals thereafter. RESULTS: Forty eyes of 35 patients were evaluated with an average follow-up of 33.6 months. Conjunctival breakdown occurred in association with 10 glaucoma drainage device implants in 9 eyes. Eleven eyes had glaucoma drainage devices placed before keratoprosthesis surgery, 3 eyes underwent glaucoma drainage device placement at the time of surgery, and 2 eyes had a glaucoma drainage device placed after surgery. All but one of the eroded glaucoma drainage devices were placed before surgery. Associated complications included endophthalmitis, hypotony, and keratoprosthesis extrusion, with 6 glaucoma drainage devices requiring removal. Long-term beset-corrected visual acuity was maintained better in eyes in which glaucoma drainage device erosions did not develop. CONCLUSIONS: One of the main challenges with keratoprosthesis surgery is treating concurrent glaucoma. Glaucoma drainage devices have been advocated as a way to address this long-term complication, but this series suggests that glaucoma drainage device-related complications can cause significant vision loss. PMID- 21636071 TI - Statins improve outcome in isolated heart valve operations: a propensity score analysis of 3,217 patients. AB - BACKGROUND: Whether statins can improve postoperative outcome in patients without coronary artery disease undergoing heart valve operations was assessed. METHODS: Data for 3,217 patients undergoing isolated valve procedures at 2 institutions between May 2003 and May 2009 were reviewed. Clinical follow-up was completed. Two propensity-matched cohorts of 1,104 patients each were identified. Multivariable regression and Kaplan-Meyer survival analysis were performed to investigate risk factors correlated with death, stroke, myocardial infarction, and cardiac arrhythmias. RESULTS: The overall 30-day mortality rate was 2.7%, and 2,096 of 2,149 hospital survivors were alive at a median follow-up of 27 months. Preoperative statin treatment was independently associated with a significant reduction in the risk of hospital death (odds ratio [OR], 0.48; 95% confidence interval [CI], 0.32 to 0.89; p=0.001), postoperative cardiac arrhythmias (OR, 0.76; 95% CI, 0.62 to 0.93; p<0.006), and stroke (OR, 0.54; 95% CI, 0.32 to 0.92; p=0.02) but was not independently associated with a reduced risk of postoperative myocardial infarction. At follow-up, Kaplan-Meyer survival analysis showed statistically significant lower rates of mortality (chi2, 4.41; hazard ratio [HR], 1.59; 95% CI, 1.13 to 2.27; p=0.03), stroke (chi2, 11.42; HR, 2.15; 95% CI, 1.37 to 3.27; p=0.0007), cardiac arrhythmias (chi2, 19.9; HR, 2.13; 95% CI, 1.81 to 2.72; p<0.0001), and major adverse cardiac and cerebrovascular events (chi2, 3.74; HR, 1.37; 95% CI, 0.99 to 1.74; p=0.05) in patients receiving statin treatment. No statistically significant difference was found between groups in myocardial infarction incidence at follow-up. CONCLUSIONS: Statin therapy is associated with a lower rate of adverse cardiovascular events after isolated heart valve operations. PMID- 21636072 TI - A user-centred approach to requirements elicitation in medical device development: a case study from an industry perspective. AB - The healthcare industry is dependent upon the provision of well designed medical devices. To achieve this it is recommended that user-centred design should begin early, and continue throughout device development. This is a challenge, particularly for smaller companies who may lack the necessary expertise and knowledge. The aim of this study was to conduct a rigorous yet focused investigation into the user requirements for a new medical imaging device. Open ended semi-structured interviews were conducted with potential clinical users of the device to investigate the clinical need for the device and the potential benefits for patients and clinical users. The study identified a number of new and significant clinical needs that suggested that the concept of the device should be fundamentally changed. The clinical and organisational priorities of the clinical users were identified, as well as a number of factors that would act as barriers to the safe and effective adoption of the device. The developers reported that this focused approach to early requirements elicitation would result in an improved product, reduce the time to market, and save the time and cost of producing and evaluating an inappropriate prototype. PMID- 21636073 TI - Increased trunk extension endurance is associated with meaningful improvement in balance among older adults with mobility problems. AB - OBJECTIVE: To determine whether trunk extension endurance changes with training are associated with clinically meaningful improvements in balance among mobility limited older adults. DESIGN: Longitudinal data from a randomized controlled trial. SETTING: Outpatient rehabilitation research center. PARTICIPANTS: Community-dwelling older adults (N=64; mean age, 75.9y) with mobility limitations as defined by a score of 4 to 10 on the Short Physical Performance Battery. INTERVENTIONS: Sixteen weeks of progressive resistance training. MAIN OUTCOME MEASURES: Outcomes were the Berg Balance Scale (BBS) and the Unipedal Stance Time (UST). Predictors included leg strength, leg power, trunk extension endurance, and the product of heart rate and blood pressure (RPP) at the final stage of an exercise tolerance test. We performed an analysis of data from participants who completed 16 weeks of training by using binary outcomes defined by a clinically meaningful change (CMC) from baseline to completion of the intervention (BBS=4 units; UST=5s). The association of predictor variables with balance outcomes was examined separately and together in multivariate adjusted logistic regression models. RESULTS: Trunk extension endurance in seconds (1.04 [1.00-1.09]) was independently associated with CMC on the BBS. Trunk extension endurance (1.02 [1.00-1.03]) was independently associated with CMC on the UST. Other physical attributes were not associated with meaningful change in balance. CONCLUSIONS: Improvements in trunk extension endurance were independently associated with CMCs in balance in older adults. Leg strength, leg power, and RPP were not associated with CMC in balance. Poor trunk extension endurance may be a rehabilitative impairment worthy of further study as a modifiable factor linked to balance among older adults. PMID- 21636074 TI - Medication, surgery, and physiotherapy among patients with the hypermobility type of Ehlers-Danlos syndrome. AB - OBJECTIVES: To describe medication use, surgery, and physiotherapy, and to examine the effect of these treatment modalities on functional impairment and amount of complaints among patients with the hypermobility type of Ehlers-Danlos syndrome (EDS-HT). DESIGN: Cross-sectional study. SETTING: Physical and rehabilitation medicine department and center for medical genetics. PARTICIPANTS: Patients with EDS-HT (N=79; 8 men, 71 women) were recruited for this study. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Patients filled out questionnaires regarding type of complaints, medication use, surgery, physiotherapy, and outcome of treatment. Functional impairment in daily life was measured by the Sickness Impact Profile. Pain severity was assessed with visual analog scales. RESULTS: Patients reported a large number of complaints, a considerable presence of severe pain, and a clinically significant impact of disease on daily functioning. Most patients (92.4%) used medications, among which analgesics were the most prevalent. Fifty-six patients (70.9%) underwent surgery, including mainly interventions of the extremities and abdomen. Forty-one patients (51.9%) are currently enrolled in a physical therapy program, mainly comprising neuromuscular exercises, massage, and electrotherapy. Patients with a high consumption of analgesics, who visited the physiotherapist, or who underwent surgery had a higher dysfunction in daily life. Only 33.9% of the patients who underwent surgery and 63.4% of patients in physical therapy reported a positive outcome. CONCLUSIONS: Patients with EDS-HT have numerous complaints and an impaired functional status that strongly determine their high rate of treatment consumption. The outcome of surgical and physiotherapy treatment is disappointing in a large percentage, which illustrates a strong need for evidence-based therapy. PMID- 21636075 TI - One plus one is less than two: visual features elicit non-additive mismatch related brain activity. AB - In a passive oddball task (performing in a video game), participants were presented with sequences of either standard stimuli or patterns containing deviant orientation, deviant spatial frequency or both deviant orientation and spatial frequency. Orientation deviants presented to the lower half of the visual field elicited a posterior negative component with a peak latency of 130 ms. Spatial frequency deviants elicited a similarly negative component that was later followed by another negative component. Activity elicited by the double-deviant stimulus was identical to activity elicited by the orientation deviant alone. The subtraction difference of the peak latency and scalp distribution of the deviant minus the standard difference potentials were unequal to those of the exogenous event-related potential (ERP) components and were therefore considered visual mismatch negativities (vMMNs). The non-additivity of the feature-related responses is interpreted as sensitivity of the implicit change-detection system to deviant events rather than an exclusive sensitivity to individual features. Deviant stimuli presented to the upper half of the field elicited responses with positive polarity, but this activity was less pronounced than the vMMN. Polarity reversal of the response to upper half-field stimulation suggests that the origin of the activity lies in retinotopic areas. Because of the emergence of a mismatch component with positive polarity, we propose that the term visual mismatch negativity (vMMN) be replaced with the more general term visual mismatch response (vMMR). PMID- 21636076 TI - Malformation of the superior olivary complex in an animal model of autism. AB - Autism is a neurodevelopmental disorder characterized by social difficulties, impaired communication skills and repetitive behavioral patterns. Additionally, there is evidence that auditory deficits are a common feature of the autism spectrum disorders. Despite the prevalence of autism, the neurobiology of this disorder is poorly understood. However, abnormalities in neuronal morphology, cell number and connectivity have been described throughout the autistic brain. Indeed, we have demonstrated significant dysmorphology in the superior olivary complex (SOC), a collection of auditory brainstem nuclei, in the autistic brain. Prenatal exposure to valproic acid (VPA) in humans has been associated with autism and in rodents prenatal VPA exposure produces many neuroanatomical and behavioral deficits associated with autism. Thus, in an effort to devise an animal model of the autistic auditory brainstem, we have investigated neuronal number and morphology in animals prenatally exposed to valproic acid (VPA). In VPA exposed rats, we find significantly fewer neurons and significant alterations in neuronal morphology. Thus, prenatal VPA exposure in rats appears to produce similar dysmorphology as we have reported in the autistic human brain. PMID- 21636077 TI - Progression of neurodegeneration and morphologic changes in the brains of juvenile mice with selenoprotein P deleted. AB - Selenoprotein P (Sepp1) is an important protein involved in selenium (Se) transport and homeostasis. Severe neurologic dysfunction develops in Sepp1 null mice (Sepp1(-/-)) fed a selenium-deficient diet. Sepp1(-/-) mice fed a selenium deficient diet have extensive degeneration of the brainstem and thalamus, and even when supplemented with selenium exhibit subtle learning deficits and altered basal synaptic transmission and short-term plasticity in the CA1 region of the hippocampus. The goal of this study was to delineate the regional progression of neurodegeneration in the brain, determine the extent of neuronal cell death, and evaluate neurite structural changes within the hippocampus of Sepp1(-/-) mice. Whole brain serial sections of wild-type and Sepp1(-/-) mice maintained on selenium-deficient or supplemented diets over the course of 12 days from weaning were evaluated with amino cupric silver neurodegeneration stain. The neurodegeneration was present in all regions upon weaning and progressed over 12 days in Sepp1(-/-) mice fed selenium-deficient diet, except in the medial forebrain bundle and somatosensory cortex where the neurodegeneration developed post-weaning. The neurodegeneration was predominantly axonal, however the somatosensory cortex and lateral striatum showed silver-stained neurons. Morphologic analysis of the hippocampus revealed decreased dendritic length and spine density, suggesting that loss of Sepp1 also causes subtle changes in the brain that can contribute to functional deficits. These data illustrate that deletion of Sepp1, and presumably selenium deficiency in the brain, produce both neuronal and axonal degeneration as well as more moderate and potentially reversible neurite changes in the developing brain. PMID- 21636078 TI - A novel highly charged exopolysaccharide produced by two strains of Stenotrophomonas maltophilia recovered from patients with cystic fibrosis. AB - Stenotrophomonas maltophilia is a non-fermenting Gram-negative microorganism capable of causing chronic pulmonary infection in cystic fibrosis patients and its ability to form biofilms on polystyrene and glass surfaces, as well as on cystic fibrosis-derived bronchial epithelial IB3-I cells was recently demonstrated. The latter evidence might explain the power of S. maltophilia to produce persistent lung infections, despite intensive antibiotic treatment. In addition to being important components of the extracellular biofilm matrix, polysaccharides are involved in virulence, as they contribute to bacterial survival in a hostile environment. With the aim of contributing to the elucidation of S. maltophilia virulence factors, the exopolysaccharides produced by two mucoid clinical isolates of S. maltophilia obtained from two cystic fibrosis patients were completely characterised, mainly by means of ESI-MS and NMR spectroscopy. The results showed that, although the two isolates were recovered from two different patients living in different countries (Italy and France), the exopolysaccharides produced have an identical primary structure, with the following repeating unit: The exopolysaccharide is highly negatively charged for the presence of three uronic acids on four residues in the repeating unit. Moreover, an ether-linked d-lactate substituent is located on C-3 and one O acetyl group on C-4 of the galacturonic acid side chain. Another O-acetyl group substitutes C-2 of the galacturonic acid in the backbone, making this primary structure unique. PMID- 21636079 TI - Increased expression of mGITRL on D2SC/1 cells by particulate beta-glucan impairs the suppressive effect of CD4+CD25+ regulatory T cells and enhances the effector T cell proliferation. AB - beta-Glucans have been shown to enhance immune responses for centuries, which contributes to their anti-tumor property. However, their mechanisms of action are still elusive. Dectin-1, the C-type lectin receptor for beta-glucan, is expressed abundantly on dendritic cells (DCs). Activation of DCs via Dectin-1 can lead to the maturation of DC, inducing both innate and adaptive immune responses against tumor development and microbial infection. In this study, we found that particulate yeast-derived beta-glucans could induce the maturation of murine dendritic cell line D2SC/1 cells and increase the expression of mGITRL on D2SC/1 cells via Dectin-1/Syk pathway in a dose dependent manner. Furthermore, we demonstrated that the increased mGITRL on D2SC/1 cells could impair the suppressive activity of CD4(+)CD25(+) regulatory T cells (Tregs) and enhance the proliferation of CD4(+)CD25(-) effector T cells (Teffs). These findings suggest that particulate beta-glucan can be used as immunomodulator to stimulate potent T cell-mediated adaptive immunity while down-regulate immune suppressive activity, leading to a more efficient defense mechanism against tumor development or infectious diseases. PMID- 21636080 TI - Experimental food allergy leads to adipose tissue inflammation, systemic metabolic alterations and weight loss in mice. AB - To investigate the consequences of food allergy in adipose tissue and metabolism, we used a murine model in which mice have been sensitized subcutaneously with ovalbumin and further received antigen-containing diet. Allergic mice presented a significant weight loss 7 days after oral challenge with a concomitant decrease in epididymal adipose tissue mass. This decrease was associated with increased lipolysis and local inflammation. In adipose tissue of allergic mice there were increased leukocyte rolling and adhesion in the microvasculature, increased number of leukocytes in the tissue, especially macrophages (F4/80(+) cells) and increased pro-inflammatory cytokines levels, including TNF-alpha, IL-6 and CCL2. In addition, we observed low serum concentrations of triglyceride, glucose, total cholesterol and free fatty acids in the allergic mice. Our results suggest that the induction of food allergy in mice leads to adipose tissue inflammation and systemic metabolic alterations that contribute to the weight loss observed. PMID- 21636081 TI - Some ethical issues in brain imaging. PMID- 21636082 TI - Organophosphate pesticide metabolite levels in pre-school children in an agricultural community: within- and between-child variability in a longitudinal study. AB - This study characterized the within- and between-child variability in dimethylthiophosphate (DMTP) levels in the urine of 44 children living in an agricultural community in central Washington State in December 1997 and 1999. The goal of this analysis was to investigate these variability components during periods when organophosphate pesticides were and were not actively applied to orchards in that community. Each child provided between 10 and 26 biweekly urine samples over a 21-month period, and these samples were analyzed for six dialkylphosphate (DAP) metabolites common to organophosphate pesticides, including DMTP. Previous analysis of this dataset found that DAP concentrations were elevated during months when organophosphate pesticides were applied to orchards in this region. The current analysis demonstrates that the within-child component of day-to-day variability was much greater than the between-child component of variability by a factor of 3-7 across the DAP metabolites that were analyzed. Therefore, organophosphate pesticide exposure appeared to vary more than 3 times from day-to-day than from child-to-child. This finding has important implications for epidemiologic and exposure pathways research, since accounting for within-child variability may increase the power of a study and allow for the detection of differences that would not otherwise be possible without an analysis that separates out the within-child variability. PMID- 21636083 TI - 'Time critical' rapid amputation using fire service hydraulic cutting equipment. AB - INTRODUCTION: Entrapped trauma victims require extrication, which, on rare occasions, may involve amputation of a limb. Standard extrication techniques sometimes fail or may be impossible, leading to the death of the entrapped victim. We propose that the use of fire service hydraulic cutting equipment can be used effectively to urgently amputate a limb, where conventional techniques are unusable. METHOD: The study aims to determine: (i) the potential use of this equipment to achieve expeditious life-saving amputations and (ii) the effect the fire service hydraulic cutting equipment has on the bony and surrounding soft tissues. Initially a porcine limb was used followed by fresh-frozen cadaveric lower limbs. We recorded the time, number of cuts, proximal fracture propagation and quality of bone cut when performing amputations at five levels. RESULTS: The experiment confirms that faster guillotine amputations in human cadaveric lower limb specimens can be achieved by using fire service hydraulic cutting equipment. Overall, the average time to complete an amputation in these ideal experimental circumstances at all five levels was quicker using the hydraulic cutting equipment. Either one or two cutting actions were required to achieve the amputation using fire service hydraulic cutting equipment. The degree and proximal extent of the comminution were greater using the fire service hydraulic cutting equipment. CONCLUSION: If circumstances and time constrains allow, a conventional amputation technique carried out by a trained medical practitioner would be preferable to the use of the fire service hydraulic cutting equipment. However, we feel that this technique could be used to perform emergent amputation under trained medical supervision, if it is felt that a standard amputation technique would take too long or the environment is too restrictive to perform a standard amputation safely. PMID- 21636084 TI - Single-stage total hip arthroplasty and fracture fixation for a both column acetabular fracture in type I osteogenesis imperfecta. PMID- 21636085 TI - Evolution of trauma care in the UK: current developments and future expectations. PMID- 21636086 TI - Monitoring micrometer-scale collagen organization in rat-tail tendon upon mechanical strain using second harmonic microscopy. AB - We continuously monitored the microstructure of a rat-tail tendon during stretch/relaxation cycles. To that purpose, we implemented a new biomechanical device that combined SHG imaging and mechanical testing modalities. This multi scale experimental device enabled simultaneous visualization of the collagen crimp morphology at the micrometer scale and measurement of macroscopic strain stress response. We gradually increased the ultimate strain of the cycles and showed that preconditioning mostly occurs in the first stretching. This is accompanied by an increase of the crimp period in the SHG image. Our results indicate that preconditioning is due to a sliding of microstructures at the scale of a few fibrils and smaller, that changes the resting length of the fascicle. This sliding can reverse on long time scales. These results provide a proof of concept that continuous SHG imaging performed simultaneously with mechanical assay allows analysis of the relationship between macroscopic response and microscopic structure of tissues. PMID- 21636087 TI - Analysis of salicylate and benzophenone-type UV filters in soils and sediments by simultaneous extraction cleanup and gas chromatography-mass spectrometry. AB - An analytical method for the determination of UV filters in soil and sediment has been developed and validated considering benzophenones (BP) and salicylates as target analytes. Soil and sediment samples were extracted with ethyl acetate methanol (90:10, v/v) assisted with sonication, performing a simultaneous clean up step. Quantification of these compounds was carried out by gas chromatography mass spectrometry (GC-MS) after derivatization of the extracts with N,O bis(trimethylsilyl)trifluoroacetamide (BSTFA). Recoveries from spiked soil samples ranged from 89.8% to 104.4% and they were between 88.4% and 105.3% for spiked sediment samples. The effect of the residence time and soil moisture content on the recovery of these compounds was also studied. The precision, expressed as relative standard deviation, was in all cases below 6.1% and the limits of detection (S/N=3) varied from 0.07 to 0.10 ng g(-1) and from 0.11 to 0.28 ng g(-1) for soils and sediments, respectively. The validated method was applied to the analysis of five benzophenone and two salicylate UV filters in soil and sediment samples collected in different areas of Spain. PMID- 21636088 TI - Application of dispersive liquid-liquid microextraction for the determination of aflatoxins B1, B2, G1 and G2 in cereal products. AB - The application of dispersive liquid-liquid microextraction (DLLME) technique for the rapid analysis of aflatoxins B(1), B(2), G(1) and G(2) in maize, rice and wheat products has been evaluated. After extraction of aflatoxins from cereal matrices with a mixture of methanol/water 8:2 (v/v), the analytes were rapidly transferred from the extract to another small volume of organic solvent, chloroform, by DLLME. Aflatoxins were determined using high performance liquid chromatography with florescence detection and photochemical post-column derivatization. Parameters affecting both extraction and DLLME procedures, such as extraction solvent, type and volume of DLLME extractant, volume of water and salt effect, were systematically investigated and optimized to achieve the best extraction efficiency. Under the optimal experimental conditions, the whole analytical method provides enrichment factors around 2.5 times and detection limits (0.01-0.17 MUg kg(-1)) below the maximum levels imposed by current regulation for aflatoxins in cereals and cereal products intended for direct human consumption. Recoveries (67-92%) and repeatability (RSD<10, n=3), tested in three different cereal matrices, meet the performance criteria required by EC Regulation No. 401/2006 for the determination of the levels of mycotoxins in foodstuffs. The proposed method was successfully applied to the analysis of retail cereal products with quantitative results comparable to the immunoaffinity chromatography (IAC). The main advantages of developed method are the simplicity of operation, the rapidity to achieve a very high sample throughput and low cost. PMID- 21636089 TI - Plunger-in-needle solid-phase microextraction with graphene-based sol-gel coating as sorbent for determination of polybrominated diphenyl ethers. AB - A solid-phase microextraction (SPME) device, assembled with a commercially available plunger-in-needle microsyringe, with the plunger coated with graphene via a sol-gel approach, was developed for the gas chromatographic-mass spectrometric determination of polybrominated diphenyl ethers (PBDEs) in environmental samples. This is the first application of graphene-based sol-gel coating as SPME sorbent. Parameters affecting the extraction efficiency were investigated in detail. The new coating exhibited enrichment factors for PBDEs between 1378 and 2859. The unique planar structure of graphene enhanced the pi-pi interaction with the aromatic PBDEs; additionally, the sol-gel coating technique created a porous three-dimensional network structure which offered larger surface area for extraction. The stainless steel plunger provided firm support for the coating and enhanced the durability of the assembly. The plunger-in-needle microsyringe represents a ready-made tool for SPME implementation. Under the optimized conditions, the method detection limits for five PBDEs were in the range of 0.2 and 5.3 ng/L (at a signal/noise ratio of 3) and the precision (% relative standard deviation, n=5) was 3.2-5.0% at a concentration level of 100 ng/L. The linearities were 5-1000 or 10-1000 ng/L for different PBDEs. Finally, the proposed method was successfully applied to the extraction and determination by gas chromatography-mass spectrometry of PBDEs in canal water samples. PMID- 21636090 TI - Characterization of several stationary phases prepared by thermal immobilization of poly(methyltetradecylsiloxane) onto silica surfaces. AB - Variations of a thermal immobilization procedure using poly(methyltetradecilsiloxane) and silica produced fourteen stationary phases with carbon contents of 4-18%. The stationary phases were chromatographically evaluated with the Engelhardt, SRM 870 and Tanaka tests. Classifications using USP and Euerby procedures indicate that the new immobilized phases are different from most commercial phases although there was some similarity with phases that have high ion-exchange interactions. The retention mechanism involved in the separation of basic solutes on several of the new stationary phases was studied by varying pH, type of Lewis base and the ionic strength of the eluent. The separations are strongly influenced by the chemistry of the accessible free silanols. The stationary phases present good selectivity at intermediate pH where the basic analytes were protonated, suggesting use of intermediate pH for these separations. Stability tests show that the stationary phases have poor stability at very high pH, even at 23 degrees C, but good stability in acidic mobile phases, even at 75 degrees C, as expected for an immobilized polymer stationary phase. PMID- 21636091 TI - Optimization of the derivatization reaction and the solid-phase microextraction conditions using a D-optimal design and three-way calibration in the determination of non-steroidal anti-inflammatory drugs in bovine milk by gas chromatography-mass spectrometry. AB - An experimental design optimization is reported of an analytical procedure used in the simultaneous determination of seven non-steroidal anti-inflammatory drugs (NSAIDs) in bovine milk by gas chromatography with mass spectrometry detection (GC-MS). This analytical procedure involves a solid-phase microextraction (SPME) step and an aqueous derivatization procedure of the NSAIDs to ethyl esters in bovine milk. The following NSAIDs are studied: ibuprofen (IBP), naproxen (NPX), ketoprofen (KPF), diclofenac (DCF), flufenamic acid (FLF), tolfenamic acid (TLF) and meclofenamic acid (MCL). Three kinds of SPME fibers - polyacrylate (PA), polydimethylsiloxane/divinylbenzene (PDMS/DVB) and polydimethylsiloxane (PDMS) - are compared to identify the most suitable one for the extraction process, on the basis of two steps: to determine the equilibrium time of each fiber and to select the fiber that provides the best figures-of-merit values calculated with three way PARAFAC-based calibration models at the equilibrium time. The best results were obtained with the PDMS fiber. Subsequently, 8 experimental factors (related to the derivatization reaction and the SPME) were optimized by means of a D optimal design that involves only 14 rather than 512 experiments in the complete factorial design. The responses used in the design are the sample mode loadings of the PARAFAC decomposition which are related to the quantity of each NSAID that is extracted in the experiment. Owing to the fact that each analyte is unequivocally identified in the PARAFAC decomposition, a calibration model is not needed for each experimental condition. The procedure fulfils the performance requirements for a confirmatory method established in European Commission Decision 2002/657/EC. PMID- 21636092 TI - Cu(II) adsorption by halloysites intercalated with sodium acetate. AB - Intercalated halloysites with sodium acetate at various contact time were prepared. The resulting materials were characterised by X-ray powder diffraction, Fourier transformed infrared spectroscopy, scanning electronic microscopy, and specific surface area evaluation. The modified halloysites were employed as Cu(II) adsorbents from aqueous solutions. Various parameters were studied through the batch method. Kinetic data, equilibrium isotherms, and thermodynamic parameters were evaluated by considering several models. The fraction of halloysite intercalated with sodium acetate remained low, up to 7 days, then linearly increased with contact time. Thirty days were required to achieve a ratio of 91%. For this sample, a proliferation of small tubes (nanotubes) was evidenced. The best results were achieved with the pseudo-second-order kinetic model associated with intraparticle diffusion and with the Redlich-Peterson isotherm, for the equilibrium data. The thermodynamic data show that adsorption would be spontaneous at low temperatures, of exothermic nature, resulting in an adsorbate-adsorbent system much more ordered. The insertion of CH(3)COONa into halloysite significantly affects the Cu(II) adsorption. The magnitude in enhancement of copper adsorption on solid phase thus depends on the content of the carboxylic functional groups, which increases with the insertion of CH(3)COONa into the halloysitic matrix. The involved mechanism is quite complex. It implies electrostatic considerations and a cationic exchange process. The most intercalated sample was found to be very effective as adsorbent of copper(II) from aqueous solutions. PMID- 21636093 TI - Layered zinc hydroxide salts: delamination, preferred orientation of hydroxide lamellae, and formation of ZnO nanodiscs. AB - Delamination of layered zinc hydroxide salts (LZH) into hydroxide layers provides nanobuilding blocs of a two-dimensional anisotropy. The methodology, extent of delamination, the size and stability of hydroxide lamellae are described in detail. The ability of lamellae to restack to form oriented hydroxide films depends on the solvent, original LZH salt, and conditions used for delamination. The most interesting results were obtained using LZH intercalated with dodecyl sulfate anions and LZH nitrate delaminated in butanol at 60 degrees C and in formamide at room temperature, respectively. The former method produces hydroxide lamellae of a lateral size of ca. 10-20 nm. The inner structure of the hydroxide layers is conserved and separated lamellae restack to the original layered structure of LZH dodecyl sulfate. The latter method yields lamellae with a size decreasing from 73.3 nm to 10 nm after a 2-week aging, while their thickness is nearly constant (2.6-3.8 nm). However, the use of formamide is complicated by the formation of Zn(II) formate. The major part of LZH intercalated with dodecyl sulfate anions is transformed during the delamination procedure to anisotropic ZnO nanoparticles, either needle-like particles prolonged in the [0 0 1] direction or disc-like particles flattened along the (0 0 1) plane. PMID- 21636094 TI - Behavioral profiles associated with auditory processing disorder and specific language impairment. AB - PURPOSE: To describe and compare behavioral profiles associated with auditory processing disorder (APD) and specific language impairment (SLI) in school-age children. METHOD: The participants in this cross-sectional observational study were 64 children (mean age 10.1 years) recruited through clinician referrals. Thirty-five participants had a clinical diagnosis of APD and 29 were receiving services for language impairment. Participants completed 18 behavioral measures of spoken language, auditory processing, reading, memory, and motor speed. Responses were used to classify children as affected/not affected with APD, and affected/not affected with SLI. Comparisons were made between children with and without an APD diagnosis, and between children assigned to the APD/not APD and SLI/not SLI groups. Agreement between clinical status and test-based classifications is also reported. RESULTS: There were no group mean differences between children with and without a clinical diagnosis of APD. Group mean differences on Cube Design and reading fluency were observed for children classified as APD/not APD; and group mean differences on nonword repetition, spatial working memory, and two auditory processing tests were observed for children classified as SLI/not SLI. CONCLUSIONS: The behavioral profiles of children with APD and SLI were very similar. Although group mean differences were found, they were difficult to interpret in terms of current theories. LEARNING OUTCOMES: The reader will be able to: (1) describe similarities and differences found between children with SLI and children with APD and (2) discuss assessment problems posed by overlapping behavioral characteristics of SLI and APD. PMID- 21636095 TI - Functional magnetic resonance imaging study of Piaget's conservation-of-number task in preschool and school-age children: a neo-Piagetian approach. AB - Jean Piaget's theory is a central reference point in the study of logico mathematical development in children. One of the most famous Piagetian tasks is number conservation. Failures and successes in this task reveal two fundamental stages in children's thinking and judgment, shifting at approximately 7 years of age from visuospatial intuition to number conservation. In the current study, preschool children (nonconservers, 5-6 years of age) and school-age children (conservers, 9-10 years of age) were presented with Piaget's conservation-of number task and monitored by functional magnetic resonance imaging (fMRI). The cognitive change allowing children to access conservation was shown to be related to the neural contribution of a bilateral parietofrontal network involved in numerical and executive functions. These fMRI results highlight how the behavioral and cognitive stages Piaget formulated during the 20th century manifest in the brain with age. PMID- 21636096 TI - Youth and young adult suicide: a study of life trajectory. AB - OBJECTIVES: Explore the unique developmental challenges and early adversity faced by youth and young adult who died of suicide. METHOD: Sixty-seven suicide victims (SG) were compared with 56 living control with no suicidal ideations in the last year, matched for age, gender, and geographical region. Mixed methods were used: consensus DSM-IV diagnoses were formulated based on Structured Clinical Interview for DSM-IV (SCID)-I and -II interviews complemented by medical charts. Life calendar method was conducted with closest third party informant. Life-history calendar served to measure life events and adversity throughout the life course and were analyzed by attributing burden of adversity score per five-year segment, which was then cluster-analyzed to define suicide victim profiles. RESULTS: During the last year, mood disorders, abuse and dependence disorders, and anxiety disorder were between 8 and 63 times more likely to be present in the suicide group. Between 0 and 4 years old, 50% of children in the SG were exposed to abuse, physical and/or sexual violence; 60% between 5 and 9 years old; and by the time they were 10-14 years old, 77% were exposed to these forms of violence. In the control group, the respective figures were 14%, 18% and 34%. In the suicide group, the trajectories leading to suicide are different as we observe two different subgroups, one with early-onset and one with later-onset of adversity. To a large extent, people in the suicide group were exposed to major adversity and they were more likely to present cumulative comorbid disorders. PMID- 21636097 TI - Trauma, dissociation, and antiretroviral adherence among persons living with HIV/AIDS. AB - BACKGROUND: There are approximately 1,000,000 persons living with HIV/AIDS (PLH) in the United States; to reduce rates of new infection and curb disease progression, adherence to HIV medication among PLH is critical. Despite elevated trauma rates in PLH, no studies to date have investigated the relationship between dissociation, a specific symptom of trauma, and HIV medication adherence. We hypothesized that Post-Traumatic Stress Disorder (PTSD) symptoms would be associated with lower adherence, and that dissociation would moderate this relationship. METHODS: Forty-three individuals with HIV were recruited from community-based clinics to participate in a cross-sectional study. The relationship of trauma, dissociation, and their interaction to the probability of antiretroviral adherence was assessed using a hierarchical binary logistic regression analysis. RESULTS: Among 38 eligible participants, greater PTSD was associated with lower odds of adherence (OR = .92, p < .05). Dissociation moderated the effect of PTSD on adherence, resulting in lower odds of adherence (OR = .95, p < .05). PTSD symptoms were significantly associated with lower odds of adherence in individuals reporting high levels of dissociation (OR = .86, p < .05) but not in those reporting low levels of dissociation (OR = 1.02, p > .05). CONCLUSIONS: This is the first study to demonstrate a relationship between dissociation and medication adherence. Findings are discussed in the context of clinical management of PLH with trauma histories and the need for interventions targeting dissociative symptomatology to optimize adherence. PMID- 21636098 TI - A double-blind, placebo-controlled, randomized, crossover pilot study of the safety and efficacy of multiple doses of intra-oral tropicamide films for the short-term relief of sialorrhea symptoms in Parkinson's disease patients. AB - OBJECTIVE: This proof-of-concept, pilot study aimed to explore the safety and anti-sialorrhea efficacy of single doses of intra-oral slow dissolving thin films containing tropicamide (NH004) or placebo. METHODS: Nineteen non-demented, idiopathic stable or fluctuating PD patients who complained of sialorrhea received 3 doses (0.3, 1, 3mg) of tropicamide and placebo in random order, separated by 7 days. A 10-cm visual analog scale (VAS) was used to measure the patient's subjective feelings of saliva levels at baseline and at 15, 30, 45, 90 and 120 min after treatment administration. For the last 7 patients, saliva volume was measured at baseline and 75 min after treatment. Fluctuating patients were evaluated in the ON-condition. RESULTS: The mean age of included patients was 67+/-12 years, 78% were male. Median disease duration was 8 years. The mean decrease in VAS score from baseline to 120 min were -0.55+/-0.54, -1.08+/-0.54, 1.53+/-0.52 and -0.81+/-0.51 for placebo and 0.3, 1 and 3mg tropicamide, respectively (F=0.6 p=0.6, ANOVA). Tropicamide 1mg resulted in a significant VAS score decrease (95%CI: -2.57 to -0.48). Saliva volume was reduced by 27%, 33% or 20% after tropicamide 0.3, 1 or 3mg vs 5% with placebo (p=0.5, Friedman). No adverse events were detected in any of the treatment sequences. DISCUSSION: Results of this pilot, proof-of-concept study show that NH004 was safe and exerted antisialorrhea effects worthy of further exploration. PMID- 21636100 TI - Primary angiitis of the central nervous system: report of eight cases from a single Italian center. AB - The primary angiitis of the central nervous system (PACNS) is a rare and potentially fatal form of vasculitis with unknown etiology. Headache and encephalopathy are the most frequent symptoms. Neuroimaging plays an important role in the diagnosis, but the pattern of abnormal findings is not specific. In some cases brain biopsy is mandatory. PACNS is often described as a diagnostic and therapeutic challenge for clinicians considering the lack of univocal diagnostic criteria and paucity of studies evaluating the long-term outcome. This brief work reports how we managed eight patients with PACNS from diagnosis to long-term follow up treatment. Headache and focal acute neurological deficits were the most common symptoms. Magnetic resonance imaging (MRI) was abnormal in all patients with different patterns of infarctions or intraparenchymal/subarachnoid hemorrhages or their combination. Cerebral angiography demonstrated pathological findings compatible with vasculitis in all cases. Other causes of encephalopathy were ruled out. Patients with severe clinical and neuroradiological findings were treated with steroids and immunosuppressive therapy using cyclophosphamide/methotrexate. Patients with less aggressive PACNS, were treated with steroids only. In one case with multiple relapses infliximab was used after first-line immunosuppressive therapy failure. PACNS suspicion is based on the combination of demographic/clinical and MRI findings and the exclusion of other causes of multifocal encephalopathy. A positive angiography has a diagnostic value in an adequate clinical field. A strict collaboration of neurologists, neuroradiologists, and immunoreumatologists is essential in the management of PACNS both in the diagnostic and therapeutic phases. PMID- 21636101 TI - Why do ischemic stroke and transient ischemic attack patients get readmitted? AB - OBJECTIVE: Readmission is an important indicator for the quality of healthcare services. The authors examined the reasons for 30-day readmission among urban stroke patients, and their clinical consequences. METHODS: Consecutive patients admitted to a JCAHO certified primary stroke center with ischemic stroke or transient ischemic attacks (TIA) were included. Demographics, TOAST mechanism, risk factors, treatments administered and discharge destination were collected. Charts were reviewed for readmissions up to 30 days from discharge. Reasons for readmission and outcomes in terms of disability and discharge destination were determined. RESULTS: Two hundred sixty-five patients (50.9% male; 79.6%African American; mean age 60.9 years) were included. There were 205(77.4%) strokes and 60(22.6%) TIAs. Thirteen (5%) patients died during their first admission. Of the remaining 252 patients, 25 (9.9%) were readmitted within 30 days. The reason for readmission was neurological in 8/25 patients (32%; 3 ischemic strokes, 1 hemorrhagic stroke and 4 TIAs); and non-neurological in 17/25 patients (68%). The frequent non-neurological reasons were infections (6/25), electrolyte disturbances (3/25) and trauma related to falls (2/25). Patients with coronary artery disease were more likely to be readmitted (45.5% vs. 14.7%; p=0.001) An NIH stroke scale >=10 predicted readmission (50.0% vs. 25.4% for NIHSS<10; p value 0.02). Patients discharged home or to acute rehabilitation units were less likely to be readmitted than those discharged to subacute rehabilitation units or nursing homes (8.2% vs. 23.8%; p value=0.01). INTERPRETATION: Disabling strokes are more likely to be readmitted. The reason is often non-neurological, and sometimes preventable. Physicians should review cases that return within 30 days and determine best practices that prevent readmission. PMID- 21636102 TI - [Recurrent respiratory infections, pneumonia and pulmonary interstitial infiltrates in a 81 year old man]. PMID- 21636099 TI - Supportive strategies to improve adherence to IFN beta-1b in multiple sclerosis- results of the betaPlus observational cohort study. AB - BACKGROUND: Low adherence to treatment in Multiple Sclerosis (MS) has been shown to lead to poor health outcomes. Various strategies to improve adherence have been suggested including educative programs, injection devices and dedicated nurse assistance. OBJECTIVE: To assess the impact of elements of the patient support program on adherence; to explore disease factors affecting adherence; and to determine whether these factors influence the choices of supportive elements. METHODS: A prospective, observational cohort study was conducted. MS patients were eligible if they had switched to Interferon beta-1b (IFNB-1b) between 1 and 3 months prior to inclusion. Data were collected at months 6, 12, 18 and 24 after inclusion. Adherence was defined as completion of both study protocol and medication at 24 months. Patients underwent evaluations of disability, quality of life, depression, and coping styles. RESULTS: A total of 1077 patients from 15 countries were included, of which 61.8% were adherent to IFNB-1b after 24-months. Depression, quality of life and autoinjector devices were baseline predictors of adherence at 24-months. Coping styles did not show to have substantial impact on adherence. Lower quality of life increased the probability of choosing supportive elements. CONCLUSION: The study showed that the usage of autoinjector devices chosen during the study was the strongest predictor of drug adherence of all the supportive elements tested in this study. PMID- 21636103 TI - Biotransformation of bufadienolides by cell suspension cultures of Saussurea involucrata. AB - The biotransformation of three bioactive bufadienolides, namely, bufotalin (1), telocinobufagin (2), and gamabufotalin (3) by cell suspension cultures of Saussurea involucrata yielded 11 products. Bufotalin yielded 3-epi-bufotalin (1a), 3-epi-desacetylbufotalin (1b), 3-epi-bufotalin 3-O-beta-D-glucoside (1c), 1beta-hydroxybufotalin (1d), and 5beta-hydroxybufotalin (1e); telocinobufagin yielded 3-dehydroscillarenin (2a), 3-dehydrobufalin (2b), and 3-epi telocinobufagin (2c); and gamabufotalin yielded 3-epi-gamabufotalin (3a), 3 dehydrogamabufotalin (3b), and 3-dehydro-Delta1-gamabufotalin (3c), respectively. Among these 11 products, 1a, 1b, 1c, 1d, 3a and 3c are previously unreported. The structures of these metabolites were elucidated based on NMR spectroscopic analyses and mass spectrometry. Most metabolites showed significant cytotoxic activities against human hepatoma (HepG2) and breast cancer (MCF-7) cell lines. In addition, the time course for the biotransformation of 3 was investigated. PMID- 21636104 TI - Associations between types of dietary fat and fish intake and risk of stroke in the Caerphilly Prospective Study (CaPS). AB - BACKGROUND: Few population-based longitudinal studies on diet and stroke have been conducted, and associations between dietary fat and fish intake and risk of stroke are unclear. OBJECTIVES: To prospectively examine relationships between intakes of total fat, saturated fat, unsaturated fat, white fish and oily fish and risk of stroke in a well-defined population of 2710 middle-aged men. STUDY DESIGN: Prospective cohort study. METHODS: Detailed information on health and lifestyle factors was collected via interview, and diet was assessed on three occasions using a food frequency questionnaire. Stroke ascertainment was by self report and inspection of clinical records. Extracted data were assessed by two independent experts. RESULTS: During a median follow-up of 18 years, 225 strokes (209 ischaemic and 19 haemorrhagic) were eligible for inclusion in the analyses. For most recent diet (i.e. food frequency questionnaire data collected immediately prior to the stroke event), there was a slightly lower risk of stroke with higher intakes of unsaturated fat and oily fish. Multiple adjusted hazard ratios (HRs) for the lowest vs highest quintiles of unsaturated fat and oily fish intakes were 0.66 [95% confidence interval (CI) 0.41-1.05, P trend = 0.13] and 0.66 (95% CI 0.41-1.05, P trend = 0.09), respectively. Baseline and cumulative diets showed a slightly higher risk of stroke with higher intake of white fish; HRs for the lowest vs highest quintiles were 1.16 (95% CI 0.76-1.77, P trend = 0.22) and 1.28 (95% CI 0.77-2.13, P trend = 0.48), respectively. CONCLUSIONS: Overall, strong associations were not found between intakes of different types of fat and fish and risk of stroke in middle-aged men. The inverse associations between unsaturated fat and oily fish intakes and risk of stroke were weak, but the direction of association was broadly consistent with other studies; however, these relatively weak associations were not conventionally statistically significant. PMID- 21636105 TI - Human metapneumovirus and human bocavirus associated with respiratory infection in Apulian population. AB - We have studied the occurrence of hBoV, hMPV and InfA-B in an Apulian population with respiratory tract infections. During influenza season 2008-2009, 116 oropharingeal swabs were collected from patients affected by Influenza-Like Illness (ILI). The PCR products of hMPV M and HBoV NP-1 genes were sequenced. 78 out of 116 samples were positive for at least one respiratory virus; hBoV was detected in 53, hMPV in 22 and InfA-B in 41 out of 116 swabs. A high rate of hBoV infection in adult (18.9%) and elderly (26.4%) subjects was found. The co infection rate was higher for hMPV (18/22 cases, 81.8%) compared to hBoV (26/53 cases, 49.1%), and InfA-B (25/41 cases, 61.0%). Co-infections were common in children. hBoV positive samples shared a high level of genetic similarity with the hBoV1 genotype, and hMPV positive samples clustered with A2 subgroup. Our results suggest that hBoV and hMPV play a role in ILI. PMID- 21636106 TI - Ozonation of secondary treated wastewater reduces ecotoxicity to Gammarus fossarum (Crustacea; Amphipoda): are loads of (micro)pollutants responsible? AB - Ozone application is an effective tool to reduce loads of (micro)pollutants in wastewater, however, its ecotoxicological implications are largely unknown. Therefore, the feeding rates of a leaf-shredding invertebrate (Gammarus fossarum) exposed to secondary (=non-ozone) or ozone treated wastewater were investigated to assess potential ecotoxicological effects. Two repetitive experiments resulted in significantly higher feeding rates for gammarids exposed to ozone compared to non-ozone treated wastewater sampled from a treatment plant equipped with a full scale ozonation. A further experiment confirmed these results also for wastewater from the same treatment plant, when ozonation was conducted at the lab-scale. However, the deviations in dissolved organic carbon profiles of ozone and non ozone wastewater did not seem to be the driving factor for the effects observed. Two additional experiments displayed on the one hand a higher feeding rate of G. fossarum if exposed to ten-fold enriched eluates from solid phase extraction cartridges loaded with ozone compared to non-ozone treated wastewater. On the other hand, the mean feeding rate of gammarids exposed to non-ozone treated wastewater, which contained hardly any (micro)pollutants (i.e. pharmaceuticals), was at the same level as wastewater from the same source additionally treated with ozone. These results suggest that not an alteration in the organic matrix but a reduction in the load of micropollutants most likely triggered the effects in the bioassay applied. Hence, the feeding rate of G. fossarum appears to be a well-suited bioassay to indicate alterations in ecotoxicological properties of wastewater due to the application of advanced oxidation processes like ozonation. PMID- 21636107 TI - Uncertainty-based calibration and prediction with a stormwater surface accumulation-washoff model based on coverage of sampled Zn, Cu, Pb and Cd field data. AB - A dynamic conceptual and lumped accumulation wash-off model (SEWSYS) is uncertainty-calibrated with Zn, Cu, Pb and Cd field data from an intensive, detailed monitoring campaign. We use the generalized linear uncertainty estimation (GLUE) technique in combination with the Metropolis algorithm, which allows identifying a range of behavioral model parameter sets. The small catchment size and nearness of the rain gauge justified excluding the hydrological model parameters from the uncertainty assessment. Uniform, closed prior distributions were heuristically specified for the dry and wet removal parameters, which allowed using an open not specified uniform prior for the dry deposition parameter. We used an exponential likelihood function based on the sum of squared errors between observed and simulated event masses and adjusted a scaling factor to cover 95% of the observations within the empirical 95% model prediction bounds. A positive correlation between the dry deposition and the dry (wind) removal rates was revealed as well as a negative correlation between the wet removal (wash-off) rate and the ratio between the dry deposition and wind removal rates, which determines the maximum pool of accumulated metal available on the conceptual catchment surface. Forward Monte Carlo analysis based on the posterior parameter sets covered 95% of the observed event mean concentrations, and 95% prediction quantiles for site mean concentrations were estimated to 470 MUg/l +/- 20% for Zn, 295 MUg/l +/- 40% for Cu, 20 MUg/l +/- 80% for Pb and 0.6 MUg/l +/- 35% for Cd. This uncertainty-based calibration procedure adequately describes the prediction uncertainty conditioned on the used model and data, but seasonal and site-to-site variation is not considered, i.e. predicting metal concentrations in stormwater runoff from gauged as well as ungauged catchments with the SEWSYS model is generally more uncertain than the indicated numbers. PMID- 21636108 TI - Virtual phosphorus ore requirement of Japanese economy. AB - Phosphorus is indispensable for agricultural production. Hence, the consumption of imported food indirectly implies the import of phosphorus resources. The global consumption of agricultural products depends on a small number of ore producing countries. For sustainable management of phosphorus resources, the global supply and demand network should be clarified. In this study, we propose the virtual phosphorus ore requirement as a new indicator of the direct and indirect phosphorus requirements for our society. The virtual phosphorus ore requirement indicates the direct and indirect demands for phosphorus ore transformed into agricultural products and fertilizer. In this study, the virtual phosphorus ore requirement was evaluated for the Japanese economy in 2005. Importantly, the results show that our society requires twice as much phosphorus ore as the domestic demand for fertilizer production. The phosphorus contained in "eaten" agricultural products was only 12% of virtual phosphorus ore requirement. PMID- 21636109 TI - Surprises perilous: toxic health hazards for employees unloading fumigated shipping containers. AB - The fumigation of freight containers to protect transported goods from fungal and pest infestation has increased worldwide in the last five years due to international regulations requiring fumigation or heat treatment of wooden packaging material and dunnage. We have found in 2008 that every sixth container and its contents do retain harmful concentrations of various fumigants and chemicals, representing a significant health risk for port and transport workers, customs officials, warehousemen, store employees and consumers. The shipping documents of these containers did not provide any information about the fumigation procedure or the used fumigant. We report here the cases of 26 patients introduced to our outpatient clinic with presumed intoxication to fumigants, or with symptoms due to inhaling the air out of fumigated containers. All patients were examined from 2007 to 2010 according to a standardized comprehensive diagnostic program. We were able to confirm the diagnosis based on typical symptoms and extensive clinical examination; by laboratory analysis we identified ethylene dichloride, methyl bromide, phosphine and methylene chloride. The predominant symptoms were headaches, concentration and memory problems, dizziness and nausea, irritation of the skin and mucous membranes and a reduced ability to do exercise. In addition to the neurological and neuropsychological impairments our analyses verified the development of reactive airways dysfunction syndrome (RADS) in 14 of 26 patients with long lasting symptoms due to their contact with fumigants. Intoxications with fumigants are serious and could be avoided. These systematical explored cases show the sustainable impact for health and socio-economic wellbeing. These findings also emphasize the necessity for international standards on permitted fumigants, appropriate labeling in the shipping documents and handling of fumigated containers. PMID- 21636110 TI - Modelling aquatic exposure and effects of insecticides--application to south eastern Australia. AB - Agricultural pesticides are widely used and can affect freshwater organisms. We applied a spatially explicit exposure model, validated for central Europe, to estimate exposure to insecticides through runoff for streams in south-eastern Australia. The model allows the identification of streams potentially affected by insecticide runoff located in 10*10 km grid cells. The computation of runoff relies on key environmental factors such as land use, soil texture, slope and precipitation. Additionally, the model predicted the ecological effect of insecticides on the macroinvertebrate community. We predicted insecticide surface runoff that results in a moderate to poor ecological quality for streams in half of the grid cells containing agricultural land. These results are in good accordance with the results obtained by estimating pesticide stress with a biotic index (SPEAR(pesticides)) based on macroinvertebrate monitoring data. We conclude that the exposure and effect model can act as an effective and cost-saving tool to identify high risk areas of insecticide exposure and to support stream management. PMID- 21636111 TI - Persistent organic pollutants in ringed seals from the Russian Arctic. AB - Organochlorine compounds total DDT (SigmaDDT), total HCH isomers (SigmaHCH), toxaphenes (sum of Parlar 26, 50, 62), mirex, endrin, methoxychlor, total chlorinated benzenes (SigmaCBz), total chlordane compounds (SigmaCHL), polychlorinated biphenyls (total of 56 congeners; SigmaPCBs), polychlorinated dibenzo-p-dioxins and furans (PCDD/Fs), and polybrominated diphenyl ethers (sum of 7 tri- to hepta congeners; SigmaPBDEs) were analysed in the blubber of adult ringed seals from the four areas of the Russian Arctic (White Sea, Barents Sea, Kara Sea and Chukchi Sea) collected in 2001-2005. Ringed seals from the south western part of the Kara Sea (Dikson Island - Yenisei estuary) were the most contaminated with SigmaDDTs, SigmaPCBs, SigmaCHL, and mirex as compared with those found in the other three areas of Russian Arctic, while the highest mean concentrations of SigmaHCHs and PCDD/Fs were found in the blubber of ringed seals from the Chukchi Sea and the White Sea, respectively. Among all organochlorine compounds measured in ringed seals from the European part of the Russian Arctic, concentrations of SigmaDDT and SigmaPCBs only were higher as compared with the other Arctic regions. Levels of all other organochlorine compounds were similar or lower than in seals from Svalbard, Alaska, the Canadian Arctic and Greenland. SigmaPBDEs were found in all ringed seal samples analysed. There were no significant differences between SigmaPBDE concentrations found in the blubber of ringed seals from the three studied areas of the European part of the Russian Arctic, while PBDE contamination level in ringed seals from the Chukchi Sea was 30-50 times lower. SigmaPBDE levels in the blubber of seals from the European part of the Russian Arctic are slightly higher than in ringed seals from the Canadian Arctic, Alaska, and western Greenland but lower compared to ringed seals from Svalbard and eastern Greenland. PMID- 21636112 TI - The nuclear factor--kappa B pathway in atherosclerosis: a potential therapeutic target for atherothrombotic vascular disease. AB - Nuclear factor kappa B (NFkappaB) is a transcription factor belonging to 'Rel' family that represents a crucial intracellular signal transduction system involved in several inflammatory diseases including atherosclerosis. Activation of NFkappaB mediated signal transduction has been established at different stages of atherosclerosis, beginning from plaque formation to its destabilization and rupture. The NFkappaB pathway is also involved in angiogenic, apoptotic and neoplastic processes. Experimental studies indicate that inhibition of these pathway may reduce inflammatory burden. The development of natural or pharmaceutical, selective and specific inhibitors of NFkappaB pathway over IkappaB kinase alpha or beta, may ultimately prove to be promising anti atherosclerotic, anti-inflammatory, antiangiogenic and antiapoptotic therapeutic instruments that could potentially reduce inflammation, attenuate atherogenesis and prevent its complications. PMID- 21636113 TI - Neoadjuvant chemotherapy and vaginal radical trachelectomy for fertility-sparing treatment in women affected by cervical cancer (FIGO stage IB-IIA1). AB - OBJECTIVES: The aim of the present report is to support the feasibility and the safety of a new fertility-sparing treatment in young women affected by bulky cervical cancer. METHODS: Between February 2007 and October 2010, seven patients presenting large IB-IIA1 tumors (30-45 mm) were scheduled for conservative treatment. All patients underwent neoadjuvant chemotherapy (NACT) followed by laparoscopic pelvic lymphadenectomy and vaginal radical trachelectomy (VRT). RESULTS: One patient presented hematological toxicity during NACT (grade 3). All patients showed complete disappearance of tumor (n=4/7) or partial response (a 50% or more decrease in total tumor size, n=3/7) to neoadjuvant treatment, and they were all treated with pelvic lymphadenectomy and VRT. Additional treatment (interstitial brachytherapy) was offered to only one woman because of a persistent parametrial tumoral lesion. After a mean follow up of 22 months (range 5-49), no relapse was observed. To date, only one woman in our study attempted to conceive and she is currently pregnant. CONCLUSIONS: Neoadjuvant chemotherapy for fertility sparing treatment is an innovative approach which is potentially quite interesting for many young women affected by bulky cervical cancer. These women, i.e. those with tumors larger than 2 cm (2-5 cm), are traditionally not offered fertility sparing treatment, thus the preliminary data we report here might have a promising impact. Nevertheless, for these patients it may be suitable to use the more radical, and time-tested, conservative surgical approach to allow for a complete and conservative excision of the residual tumor after neoadjuvant treatment. Studies with a larger number of patients and adequate follow-up are required to validate this conservative approach and to define clearly the good indications for this treatment. PMID- 21636114 TI - Risk factors for lymph node metastasis in apparent early-stage epithelial ovarian cancer: implications for surgical staging. AB - OBJECTIVES: The extent of lymphadenectomy to be performed in apparent early-stage epithelial ovarian cancer (EOC) is not well defined. We evaluated the patterns of lymphatic spread in apparent early-stage EOC and risk factors for lymph node metastasis, as these have potential implications for clinical decision making. METHODS: All cases of apparent early-stage EOC diagnosed at our institution between January 1994 and December 2003 were retrospectively identified. Apparent early-stage EOC was defined as gross disease that appeared confined to the pelvis without abdominal spread at the time of initial exploration. Demographics, pathologic findings, staging procedures performed, and clinical impression at surgery were analyzed. Patterns of lymph node positivity and risk factors associated with upstaging were assessed. RESULTS: One hundred and ninety patients with apparent early-stage EOC undergoing primary surgical staging met criteria for inclusion. All patients had at least some pathologic assessment of lymph nodes, with 115 having both bilateral pelvic and paraaortic lymphadenectomy performed. After review of pathology and operative reports, the final FIGO staging within the cohort was 54 IA (28.4%), 10 IB (5.3%), 51 IC (26.8%), 1 IIA (0.5%), 4 IIB (2.1%), 37 IIC (19.5%), 8 IIIA (4.2%), 25 IIIC (13.2%). Overall 25/190 (13%) had lymph nodes metastasis as follows: 8 (32%) had positive pelvic nodes, 12 (48%) had positive paraaortic nodes, and 5 (20%) had both positive pelvic and paraaortic lymph nodes. Significant risk factors for lymph node metastasis included bilateral vs. unilateral primary lesion (26.8% vs. 7.5%, p<0.001), positive cytologic washings vs. negative (22.4% vs. 9.1%, p=0.012), ascites vs. no ascites (28.2% vs. 9.3%, p=0.002), serous vs. other histology (28% vs. 9%, p=0.001), grade 1 vs. grade 2 vs. grade 3 disease (2.7% vs. 1.9% vs. 23.2%, p<0.001), and preoperative CA 125 levels of >35 vs. <= 35 U/ml (22.4% vs.0% p=0.006). No patients with mucinous cancers (n=29) had lymph node metastases. Patterns of LN metastases were largely independent of laterality of primary lesions: among those with unilateral lesions and positive nodes (n=10), 5 (50%) had ipsilateral lymph node involvement, 4 (40%) had bilateral involvement, and 1 (10%) had isolated contralateral lymph nodes positive. CONCLUSIONS: Complete surgical staging in EOC patients with gross disease confined to the ovaries and pelvis should include bilateral pelvic and paraaortic lymphadenectomy. Even in patients with unilateral lesions, lymph node metastases are commonly bilateral. Bilateral ovarian lesions, positive cytology, presence of ascites, high grade histology, and serous histology are risk factors for lymph node involvement. This information may be helpful in counseling patients presenting for consideration of re-staging after unexpected findings of malignancy. PMID- 21636115 TI - Downregulation of basophil-derived IL-4 and in vivo T(H)2 IgE responses by serotonin and other organic cation transporter 3 ligands. AB - BACKGROUND: Murine basophils can contribute to the T(H)2 polarization of the immune response by providing rapidly large amounts of IL-4, which suggests that pharmacologic downregulation of this cytokine might provide a strategy to attenuate pathologies associated with excessive production. OBJECTIVE: We examined a number of physiological and pharmacologic ligands of the organic cation transporter 3 (OCT3), a membrane carrier of biogenic amines, for their inhibitory effect on IL-4 production by basophils, selecting the most efficient compounds for in vivo evaluation in basophil-dependent experimental models. METHODS: IL-4 production by basophils isolated ex vivo or from bone marrow cultures was assessed in response to various stimuli with or without biogenic monoamines or pharmacologic analogs. Selected compounds were administered in vivo to examine their effect on levels of circulating IgE generated during a basophil dependent T(H)2 response and on basophil activation in mice receiving IL-33. RESULTS: We found a drastic decrease in IL-4 production by stimulated basophils on exposure to serotonin (5-hydroxytryptamine [5-HT]) that is taken up by basophils through the specific high-affinity transporters serotonin transporter and the polyspecific, high-capacity organic cation transporter 3 (OCT3; or Slc22a3) but inhibits their function exclusively through the latter. This downregulation is likewise observed in vivo in response to 5-HT and other OCT3 ligands, as well as in human basophils sorted from PMBCs of nonatopic donors. CONCLUSIONS: We provide evidence for a new means of downregulating IL-4 production by basophils, both in vitro and in vivo, through OCT3 targeted by 5-HT and pharmacologic ligands. PMID- 21636116 TI - Efficacy and safety of omalizumab in patients with chronic urticaria who exhibit IgE against thyroperoxidase. AB - BACKGROUND: A subgroup of patients with chronic spontaneous urticaria (CU) exhibits IgE antibodies directed against autoantigens, such as thyroperoxidase (TPO). We conducted this study to investigate whether such patients with CU with IgE against TPO benefit from treatment with omalizumab, a humanized anti-IgE mAb licensed for the treatment of severe persistent allergic (IgE-mediated) asthma. OBJECTIVES: We sought to assess the efficacy of omalizumab treatment in patients with CU with IgE autoantibodies against TPO. METHODS: In this multicenter, randomized, double-blind, placebo-controlled study patients with CU (male/female, 18-70 years of age) with IgE autoantibodies against TPO who had persistent symptoms (wheals and pruritus) despite standard antihistamine therapy were randomized to receive either omalizumab (75-375 mg, dose determined by using the approved asthma dosing table) or placebo subcutaneously once every 2 or 4 weeks for 24 weeks. The primary end point was the change from baseline in mean weekly urticaria activity score after 24 weeks of treatment, as calculated from patients' diaries. The safety and tolerability of omalizumab were also assessed. RESULTS: Of the 49 randomized patients (omalizumab, n = 27; placebo, n = 22), 42 completed the study. At week 24, patients demonstrated a mean reduction in the weekly urticaria activity score from baseline of 17.8 with omalizumab and 7.9 with placebo (P = .0089). Complete protection from wheal development was observed in 19 (70.4%) patients in the omalizumab group compared with only 1 (4.5%) patient in the placebo group. The rate of adverse events was similar in both groups. CONCLUSIONS: The results of this study indicate that omalizumab is an effective treatment option for patients with CU with IgE autoantibodies against TPO who are refractory to conventional treatment. PMID- 21636117 TI - Long-term outcomes in pediatric-onset esophageal eosinophilia. AB - BACKGROUND: Pediatric eosinophilic esophagitis (EoE) is a newly recognized antigen-induced form of chronic esophagitis (CE). OBJECTIVE: Characterization of long-term clinical outcomes in patients with pediatric EoE is needed. METHODS: From histologic review of 3817 pediatric esophageal biopsy specimens from 1982 1999, we conducted a nested case-control study of patients with retrospectively identified histologic eosinophilic esophagitis (rEoE) and CE, as well as an age matched control cohort. Participants were asked to complete validated health related outcome questionnaires. RESULTS: At an average of 15 years after initial endoscopy, both cohorts (42/198 patients with rEoE and 67/468 patients with CE, as well as 100 age-matched control subjects) completed questionnaires. Compared with control subjects, quality of life was significantly decreased among patients with rEoE (P < .001) and patients with CE (P < .001). Rates of dysphagia (patients with rEoE, 49%; patients with CE, 37%; control subjects, 6%) and food impaction (patients with rEoE, 40%; patients with CE, 14%; control subjects, 3%) were significantly increased in the rEoE cohort compared with those seen in control subjects (P < .001 and P < .001, respectively). Increased esophageal eosinophil counts (odds ratio [OR], 1.6; 95% CI, 1.1-2.5; P < .05) during childhood were predictive of dysphagia during early adulthood. Food allergy (OR, 2.7; 95% CI, 1.2-6.0; P < .01), allergic rhinitis (OR, 3.5; 95% CI, 1.8-6.8; P < .001), and asthma (OR, 2.1; 95% CI, 1.04-4.3; P = .04) were associated with dysphagia. Food impaction was more common among patients with reported food allergy than among those without (OR, 3.1; 95% CI, 1.2-7.8; P = .02). CONCLUSIONS: Esophageal eosinophilia is associated with reduced quality of life and persistent symptoms 15 years after presentation. Increased esophageal eosinophil counts and the occurrence of food allergy and atopy in childhood increase the rate of dysphagia in young adulthood. PMID- 21636119 TI - Remodeling in asthma. AB - Airway remodeling encompasses the structural alterations in asthmatic compared with normal airways. Airway remodeling in asthmatic patients involves a wide array of pathophysiologic features, including epithelial changes, increased smooth muscle mass, increased numbers of activated fibroblasts/myofibroblasts, subepithelial fibrosis, and vascular changes. Multiple cytokines, chemokines, and growth factors released from both inflammatory and structural cells in the airway tissue create a complex signaling environment that drives these structural changes. However, recent investigations have changed our understanding of asthma from a purely inflammatory disease to a disease in which both inflammatory and structural components are equally involved. Several reports have suggested that asthma primarily develops because of serious defects in the epithelial layer that allow environmental allergens, microorganisms, and toxins greater access to the airway tissue and that can also stimulate the release of mediators from the epithelium, thus contributing to tissue remodeling. Lung-resident fibroblasts and smooth muscle cells have also been implicated in the pathogenesis of airway remodeling. Remodeling is assumed to result in persistent airflow limitation, a decrease in lung function, and airway hyperresponsiveness. Asthmatic subjects experience an accelerated decrease in lung function compared with healthy subjects, which is proportionally related to the duration and severity of their disease. PMID- 21636120 TI - Tiotropium improves lung function in patients with severe uncontrolled asthma: a randomized controlled trial. AB - BACKGROUND: Some patients with severe asthma remain symptomatic and obstructed despite maximal recommended treatment. Tiotropium, a long-acting inhaled anticholinergic agent, might be an effective bronchodilator in such patients. OBJECTIVE: We sought to compare the efficacy and safety of 2 doses of tiotropium (5 and 10 MUg daily) administered through the Respimat inhaler with placebo as add-on therapy in patients with uncontrolled severe asthma (Asthma Control Questionnaire score, >= 1.5; postbronchodilator FEV1, <= 80% of predicted value) despite maintenance treatment with at least a high-dose inhaled corticosteroid plus a long-acting beta2-agonist. METHODS: This was a randomized, double-blind, crossover study with three 8-week treatment periods. The primary end point was peak FEV1 at the end of each treatment period. RESULTS: Of 107 randomized patients (54% female patients; mean, 55 years of age; postbronchodilator FEV1, 65% of predicted value), 100 completed all periods. Peak FEV1 was significantly higher with 5 MUg (difference, 139 mL; 95% CI, 96-181 mL) and 10 MUg (difference, 170 mL; 95% CI, 128-213 mL) of tiotropium than with placebo (both P < .0001). There was no significant difference between the active doses. Trough FEV1 at the end of the dosing interval was higher with tiotropium (5 MUg: 86 mL [95% CI, 41 132 mL]; 10 MUg: 113 mL [95% CI, 67-159 mL]; both P < .0004). Daily home peak expiratory flow measurements were higher with both tiotropium doses. There were no significant differences in asthma-related health status or symptoms. Adverse events were balanced across groups except for dry mouth, which was more common on 10 MUg of tiotropium. CONCLUSION: The addition of once-daily tiotropium to asthma treatment, including a high-dose inhaled corticosteroid plus a long-acting beta2 agonist, significantly improves lung function over 24 hours in patients with inadequately controlled, severe, persistent asthma. PMID- 21636118 TI - Lung imaging in asthmatic patients: the picture is clearer. AB - Imaging of the lungs in patients with asthma has evolved dramatically over the last decade with sophisticated techniques, such as computed tomography, magnetic resonance imaging, positron emission tomography, and single photon emission computed tomography. New insights into current and future modalities for imaging in asthmatic patients and their application are discussed to potentially shed a clearer picture of the underlying pathophysiology of asthma, especially severe asthma, and the proposed clinical utility of imaging in patients with this common disease. PMID- 21636121 TI - Glucocorticoid treatment in community-acquired pneumonia. PMID- 21636123 TI - The enhancement of mature vessel formation and cardiac function in infarcted hearts using dual growth factor delivery with self-assembling peptides. AB - For successful treatment of myocardial infarction (MI), it is important to prevent cardiac fibrosis and maintain cardiac function by protecting cardiomyocytes and inducing angiogenesis. To establish functional and stable vessels, various growth factors, ones stimulating both endothelial cells (EC) and vascular smooth muscle cells (VSMC), are required. Self-assembling peptides form fibers (<10 nm) and provide 3-dimensional microenvironments that can recruit EC and VSMC to promote vascularization and long-term delivery of growth factors. Here we demonstrate myocardial protection of infarcted heart using dual growth factor delivery with self-assembling peptides. After coronary artery ligation in rats, growth factors (PDGF-BB and FGF-2) with self-assembling peptides were injected. There were 6 rats in each group. Hearts were harvested at 4 and 8 weeks for functional and histological analysis. Infarct size and cardiomyocyte apoptosis in dual growth factors along with self-assembling peptides group were dramatically reduced compared to sham. The capillary and arterial density of this group recovered with angiogenic synergism and cardiac functions had almost recovered. In conclusion, dual growth factors along with self-assembling peptides lead to myocardial protection, stable vessel formation, and improvement in cardiac function. PMID- 21636122 TI - Dexamethasone and length of hospital stay in patients with community-acquired pneumonia: a randomised, double-blind, placebo-controlled trial. AB - BACKGROUND: Whether addition of corticosteroids to antibiotic treatment benefits patients with community-acquired pneumonia who are not in intensive care units is unclear. We aimed to assess effect of addition of dexamethasone on length of stay in this group, which might result in earlier resolution of pneumonia through dampening of systemic inflammation. METHODS: In our double-blind, placebo controlled trial, we randomly assigned adults aged 18 years or older with confirmed community-acquired pneumonia who presented to emergency departments of two teaching hospitals in the Netherlands to receive intravenous dexamethasone (5 mg once a day) or placebo for 4 days from admission. Patients were ineligible if they were immunocompromised, needed immediate transfer to an intensive-care unit, or were already receiving corticosteroids or immunosuppressive drugs. We randomly allocated patients on a one-to-one basis to treatment groups with a computerised randomisation allocation sequence in blocks of 20. The primary outcome was length of hospital stay in all enrolled patients. This study is registered with ClinicalTrials.gov, number NCT00471640. FINDINGS: Between November, 2007, and September, 2010, we enrolled 304 patients and randomly allocated 153 to the placebo group and 151 to the dexamethasone group. 143 (47%) of 304 enrolled patients had pneumonia of pneumonia severity index class 4-5 (79 [52%] patients in the dexamethasone group and 64 [42%] controls). Median length of stay was 6.5 days (IQR 5.0-9.0) in the dexamethasone group compared with 7.5 days (5.3-11.5) in the placebo group (95% CI of difference in medians 0-2 days; p=0.0480). In hospital mortality and severe adverse events were infrequent and rates did not differ between groups, although 67 (44%) of 151 patients in the dexamethasone group had hyperglycaemia compared with 35 (23%) of 153 controls (p<0.0001). INTERPRETATION: Dexamethasone can reduce length of hospital stay when added to antibiotic treatment in non-immunocompromised patients with community-acquired pneumonia. FUNDING: None. PMID- 21636124 TI - Controlled release of DNA from poly(vinylpyrrolidone) capsules using cleavable linkers. AB - The design of polymer carriers with tunable degradation and cargo release is fundamental for applications in drug and gene delivery. In this study, we report low-fouling poly(N-vinyl pyrrolidone) (PVPON) capsules assembled via hydrogen bonding and stabilized using covalent cross-linking. We first investigated the effects of pH and ionic strength to optimize the assembly conditions. A model therapeutic cargo (plasmid DNA) was then loaded in the capsules and used for encapsulation and release studies. Two bisazide cross-linkers that contain a disulfide bond, termed PEG8 (poly(ethylene glycol)) and PEG(16), were employed to stabilize the multilayer films, and used to tune the degradation and cargo release behavior of the capsules in simulated cytoplasmic conditions. The results suggest that PEG8-stabilized capsules were more efficiently cross-linked, and hence displayed higher plasmid encapsulation. Consequently, the capsules cross linked with PEG8 also showed a two-fold reduction in degradation rate. This ability to achieve controlled carrier degradation and cargo release makes these capsules of potential interest for drug and gene delivery. PMID- 21636125 TI - A gene delivery system for human cells mediated by both a cell-penetrating peptide and a piggyBac transposase. AB - The piggyBac (PB) transposable element has recently accumulated enormous attention as a tool for the transgenesis in various eukaryotic organisms. Arginine-rich cell-penetrating peptides (CPPs) are protein transduction domains containing a large amount of basic amino acids that were found to be capable of delivering biologically active macromolecules into living cells. In this study, we demonstrate a strategy, which we called "transposoduction", which is a one plasmid gene delivery system mediated by the nontoxic CPP-piggyBac transposase (CPP-PBase) fusion protein to accomplish both protein transduction and transposition. CPPs were proven to be able to synchronously deliver covalently linked PBase and noncovalently linked a cis plasmid into human cells. The expression of promoterless reporter genes coding for red (dTomato) and yellow (mOrange) fluorescent proteins (RFP and YFP) with PB elements could be detected in cells treated with the PBase-expressing plasmid after 3 days indicating transposition of coding regions to downstream of endogenous promoter sequences. An enhanced green fluorescent protein (EGFP) plasmid-based excision assay further confirmed the efficiency of the bifunctional CPP-PBase fusion protein. In conclusion, this strategy representing a combinational concept of both protein transduction and mobile transposition may provide tremendous potential for safe and efficient cell line transformation, gene therapy and functional genomics. PMID- 21636126 TI - Functional fibrils derived from the peptide TTR1-cycloRGDfK that target cell adhesion and spreading. AB - Peptide self-assembly offers a route for the production of fibrous nanomaterials with advanced bioactive properties that promote specific cell interactions. In this study the peptide TTR1-cycloRGDfK was designed to form amyloid-like fibrils that display the functional cyclic RGDfK pentapeptide ligand to target mammalian cell surface alpha(V)beta3 integrin receptors. The TTR105-115 (or TTR1) sequence was used as the self-assembling domain. Once assembled, TTR1-cycloRGDfK fibrils display a characteristic cross-beta core structure by X-ray fibre diffraction that was preserved following dehydration. Thin films of fibrils were characterised by infrared synchrotron mapping, scanning electron microscopy and atomic force microscopy. Cell adhesion and spreading were promoted on thin films of TTR1-cycloRGDfK fibrils via specific interactions with the cyclic RGDfK ligand. Low levels of non-specific interactions were also observed between cells and non-functionalised fibrils. TTR1-cycloRGDfK fibrils are an advance on bioactive fibrils previously designed to interact with a range of RGD binding integrins and our findings show that the assembly of amyloid-like fibrils based on the TTR1 sequence is robust and can be directed to form materials with specific properties. PMID- 21636127 TI - Guided sprouting from endothelial spheroids in fibrin gels aligned by magnetic fields and cell-induced gel compaction. AB - An aligned engineered microvascular network is critical to the culture of thick or highly metabolic tissue in vitro due to the need for inlet and outlet sides for perfusion of the network. Contact guidance may be a way to achieve aligned networks, but the relationship between the alignment of endothelial sprouts and the alignment of extracellular matrix fibers has yet to be fully elucidated. The data presented here show that sprouts from human blood outgrowth endothelial cell spheroids align with fibrin fibrils, and that the extent to which the sprouts align depends upon the strength of the fibril alignment. This was true for both magnetically-aligned fibrin and fibrin aligned via cell-induced gel compaction, although magnetically-aligned fibrin was more effective over the same culture period. The data also demonstrate that longer sprouts are grown when the fibrils, and thus the sprouts, are more strongly aligned. The formation of aligned endothelial sprouts using these methods can be an essential step in the generation of aligned microvascular networks. PMID- 21636128 TI - Carbon nanotube nanoreservior for controlled release of anti-inflammatory dexamethasone. AB - On demand release of anti-inflammatory drug or neurotropic factors have great promise for maintaining a stable chronic neural interface. Here we report the development of an electrically controlled drug release system based on conducting polymer and carbon nanotubes. Drug delivery research using carbon nanotubes (CNTs) has taken advantage of the ability of CNTs to load large amounts of drug molecules on their outer surface. However, the utility of the inner cavity of CNTs, which can increase the drug loading capacity, has not yet been explored. In this paper, the use of multi-wall CNTs as nanoreserviors for drug loading and controlled release is demonstrated. The CNTs are pretreated with acid sonication to open their ends and make their outer and inner surfaces more hydrophilic. When dispersed and sonicated in a solution containing the anti-inflammatory drug dexamethasone, experiments show that the pretreated CNTs are filled with the drug solution. To prevent the unwanted release of the drug, the open ends of the drug filled CNTs are then sealed with polypyrrole (PPy) films formed through electropolymerization. The prepared electrode coating significantly reduced the electrode impedance, which is desired for neural recording and stimulation. More importantly, the coating can effectively store drug molecules and release the bioactive drug in a controlled manner using electrical stimulation. The dexamethasone released from the PPy/CNT film was able to reduce lipopolysaccharide induced microglia activation to the same degree as the added dexamethasone. PMID- 21636129 TI - Robust CNS regeneration after complete spinal cord transection using aligned poly L-lactic acid microfibers. AB - Following spinal cord injury, axons fail to regenerate without exogenous intervention. In this study we report that aligned microfiber-based grafts foster robust regeneration of vascularized CNS tissue. Film, random, and aligned microfiber-based conduits were grafted into a 3 mm thoracic rat spinal cord gap created by complete transection. Over the course of 4 weeks, microtopography presented by aligned or random poly-L-lactic acid microfibers facilitated infiltration of host tissue, and the initial 3 mm gap was closed by endogenous cell populations. This bulk tissue response was composed of regenerating axons accompanied by morphologically aligned astrocytes. Aligned fibers promoted long distance (2055 +/- 150 MUm), rostrocaudal axonal regeneration, significantly greater than random fiber (1162 +/- 87 MUm) and film (413 +/- 199 MUm) controls. Retrograde tracing indicated that regenerating axons originated from propriospinal neurons of the rostral spinal cord, and supraspinal neurons of the reticular formation, red nucleus, raphe and vestibular nuclei. Our findings outline a form of regeneration within the central nervous system that holds important implications for regeneration biology. PMID- 21636130 TI - Mediation of osteogenic differentiation of human mesenchymal stem cells on titanium surfaces by a Wnt-integrin feedback loop. AB - Peri-implant bone formation depends on the ability of mesenchymal cells to colonize the implant surface and differentiate into osteoblasts. Human mesenchymal stem cells (HMSCs) undergo osteoblastic differentiation on microstructured titanium (Ti) surfaces in the absence of exogenous factors, but the mechanisms are unknown. Wnt proteins are associated with an osteoblast phenotype, but how Wnt signaling regulates HMSC differentiation on microstructured Ti surfaces is not known. HMSCs were cultured on tissue culture polystyrene or Ti (PT [Sa = 0.33 MUm, theta = 96 degrees ], SLA [Sa = 2.5 MUm, theta = 132 degrees ], modSLA [hydrophilic-SLA]). Expression of calcium-dependent Wnt ligand WNT5A increased and canonical Wnt pathway ligands decreased on microstructured Ti in a time-dependent manner. Treatment of HMSCs with canonical ligand Wnt3a preserved the mesenchymal phenotype on smooth surfaces. Treatment with Wnt5a increased osteoblastic differentiation. Expression of integrins ITGA1, ITGA2, and ITGAV increased over time and correlated with increased WNT5A expression. Treatment of HMSCs with Wnt5a, but not Wnt3a, increased integrin expression. Regulation of integrin expression due to surface roughness and energy was ablated in WNT5A-knockdown HMSCs. This indicates that surface properties regulate stem cell fate and induce osteoblast differentiation via the Wnt calcium dependent pathway. Wnt5a enhances osteogenesis through a positive feedback with integrins and local factor regulation, particularly though BMP signaling. PMID- 21636131 TI - The effects of intramuscular and intraperitoneal injections of benzo[a]pyrene on selected biomarkers in Clarias gariepinus. AB - This study investigated the dose-dependent and time-course effects of intramuscular (i.m.) and intraperitoneal (i.p.) injection of benzo[a]pyrene (BaP) on the biomarkers EROD activity, GST activity, concentrations of BaP metabolites in bile, and visceral fat deposits (Lipid Somatic Index, LSI) in African catfish (Clarias gariepinus). Intraperitoneal injection resulted in 4.5 times higher accumulation of total selected biliary FACs than i.m. injection. Hepatic GST activities were inhibited by BaP via both injection methods. Dose-response relationships between BaP injection and both biliary FAC concentrations and hepatic GST activities were linear in the i.p. injected group but nonlinear in the i.m. injected fish. Hepatic EROD activity and LSI were not significantly affected by BaP exposure by either injection route. We conclude that i.p. is a more effective route of exposure than i.m. for future ecotoxicological studies of PAH exposure in C. gariepinus. PMID- 21636132 TI - Carryover of cigarette smoke effects on hematopoietic cytokines to F1 mouse litters. AB - Neutrophils have been implicated in the pathogenesis of COPD, being recruited into the lung in response to cigarette smoke (CS) inhalation and responsible for the release of proteases and oxidant-producing enzymes, resulting in bronchitis and emphysema. Several hematopoietic cytokines are involved in neutrophil growth and recruitment; however, little is known about the effects of CS on hematopoietic cytokines are transmitted between generations. In the present investigation we evaluate the expression of hematopoietic and proinflammatory cytokines in different organs of female F(0) mice subjected to sub-chronic CS exposure, and in F(1) litters. Virgin female Balb/c mice inhaled either air or air containing CS for 90 days. The specific resistance of the airways (sRaw) was evaluated and, thereafter, the mice were mated with unexposed adult males. The levels of granulocyte-macrophage colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF), interleukin-6 (IL-6), IL-1beta and TNF-alpha mRNA and protein were evaluated in the bone marrow, amniotic fluid and bronchoalveolar lavage fluid (BALF) of F(0) dams at gestation day(14) (gd(14)) and the bone marrow, BALF and lungs of F(0) dams and F(1) littermates at post natal day(21) (pnd(21)). At gd(14), overexpression of GM-CSF, G-CSF and IL-6 mRNA and protein was observed in the bone marrow, amniotic fluid and BALF of F(0) dams. These hematopoietic cytokines were also overexpressed in the lungs of F(1) littermates compared with the control F(1) litters at pnd(21). Lineage-specific hematopoietic growth factors may play an important role in the transmission of neutrophil-associated disease susceptibility across generations. PMID- 21636133 TI - Psychological factors associated with the incidence and persistence of suicidal ideation. AB - BACKGROUND: Suicidal ideation has been identified as both a common antecedent and a significant risk factor for suicide attempt and completed suicide. However, little is known about the incidence and persistence of suicidal ideation in the general population and the associated risk factors. METHODS: A 12-month follow-up survey investigated 997 of the respondents who participated in the baseline territory-wide survey of adult population in Hong Kong. A set of baseline psychological factors was considered as predictors of first onset and persistence of suicidal ideation. RESULTS: Twelve-month incidence (1.9%) and persistence (6.2%) rates were estimated. Respondents with anxiety and lack of reasons for living were more likely to report a development of suicidal thoughts in the follow-up assessment, while respondents with higher level of average life distress and lower level of hope were at increased risk of continuing to have suicidal thoughts. Depression was found to partially mediate the effect of average life distress on persistent suicidality. LIMITATIONS: Retention rate of the follow-up sample was about 50% only. Assessments of suicidal ideation were based on retrospective reports. CONCLUSIONS: Psychological factors differentially predict first onset and persistence of suicidal ideation. It is of clinical value that depression partially mediated the effect of life distress on persistence of suicidality. PMID- 21636135 TI - Varenicline prevents affective and cognitive exacerbation during smoking abstinence in male patients with schizophrenia. AB - To explore the effects of varenicline on the psychopathology and cognition of chronic inpatients with schizophrenia, we conducted a non-randomized control group time series investigation between March 2009 and April 2010. In a mandatory smoking cessation intervention, 41 male inpatient smokers were scheduled to undergo either a 5-week varenicline treatment (varenicline group) or the use of no drugs (non-treatment group). Depression (HAM-D), anxiety (HAM-A), and psychosis (PANSS) were evaluated at baseline, and at the 2nd, 4th, 8th and 12th week after abstinence; four neuropsychological tests, including Digit Span Forward and Backward (DSF and DSB), and Trail Making Test-A and -B, were evaluated at baseline and at the 4th, 8th and 12th week. .Thirty patients completed the study. Among 15 patients in the non-treatment group, the HAM-D, HAM A, DSF, and DSB scores were exacerbated during the 2-8 weeks of abstinence, but there were no changes in psychotic symptoms and the other two neuropsychological tests. Compared with the non-treatment group, varenicline users experienced less impairment in HAM-D and HAM-A scores at the 2nd and 4th weeks, and in DSF tasks at the 4th week after abstinence. In conclusions, varenicline can attenuate abstinence-induced adverse outcomes and appears to be well-tolerated in smokers with schizophrenia. PMID- 21636134 TI - Olanzapine vs. risperidone in patients with first-episode schizophrenia and a lifetime history of cannabis use disorders: 16-week clinical and substance use outcomes. AB - The purpose of this study is to compare the efficacy of olanzapine and risperidone for the acute treatment of first-episode schizophrenia patients with cannabis use disorders. This secondary analysis of a previously published study included 49 first-episode patients with a diagnosis of schizophrenia, schizophreniform disorder, or schizoaffective disorder and a co-occurring lifetime diagnosis of cannabis use disorders randomly assigned to treatment with either olanzapine (n=28) or risperidone (n=21) for 16weeks. The olanzapine group did not differ significantly from the risperidone group for initial response rates of positive symptoms, and rates of cannabis use or alcohol use during the study. Positive symptoms and the Scale for Assessment of Negative Symptoms (SANS) global asociality-anhedonia scores improved over time but did not differ between study medications. In both groups, cannabis use during the study was higher in patients who used cannabis within three months of the admission. Thus, our results suggest that olanzapine and risperidone had a similar initial efficacy on psychotic symptoms and substance use in first-episode patients with co-occurring cannabis use disorders. If clinicians are choosing between olanzapine versus risperidone treatment for this population, their decision should be based upon factors other than symptom response and short-term substance misuse. PMID- 21636136 TI - A systematic review of the effect of tympanostomy tubes in children with recurrent acute otitis media. AB - OBJECTIVE: Documentation of the effect of tympanostomy tubes in children with recurrent acute otitis media (RAOM) is limited. A recently published Cochrane review on the effect of tympanostomy tubes in children with RAOM was based on only two studies. Could the documentation be increased by including other randomized studies? METHODS: A MEDLINE and EMBASE search for randomized controlled trials was performed and 143 eligible papers were found. Only five studies could be included. All five were randomized studies with a total of 519 children, four randomized by children and one by ears. All five studies had different designs and control groups, making a proper meta-analysis impossible. Three studies had an antibiotic treated group, two studies a placebo group, and two studies a no treatment group as comparison group. Outcome measures were rates of AOM or fraction free of AOM in six or 12 months. RESULTS: Between two and five children have to be treated with tympanostomy tubes to prevent one child from attacks of acute otitis media (AOM) in six months. Tube treatment could reduce AOM with about one attack in six months after operation. Six months treatment with antibiotics was not different from treatment with tubes. No study reported quality of life for child and family or parental absence from day care or work. CONCLUSION: Insertion of tympanostomy tubes or long-term treatment with antibiotics seems to prevent one attack of AOM or keep one child out of three free from AOM in six months. PMID- 21636137 TI - Depressive symptoms, inflammatory markers and body composition in elderly with and without chronic obstructive pulmonary disease (COPD). AB - Our aim was to assess the relationships between cortisol, interleukin-2 (Il-2) and tumor necrosis factor-alpha (TNF-alpha) levels in elderly with and without COPD presenting with or without depressive symptoms. Forty COPD patients and 53 elderly individuals with no COPD took part in the study. Depressive symptoms (Geriatric Depression Scale=GDS-15), IL-2 and TNF-alpha, serum cortisol, number of comorbidities, smoking habits and body composition were evaluated. The prevalence of depressive symptoms was higher in COPD group. The number of comorbidities was higher in patients with depressive symptoms. No differences were found between IL-2, TNF-alpha and cortisol levels, years of smoking and smoked pack-years in the groups. The COPD group obtained lower body mass index (BMI) and fat content and higher fat free mass index as well as greater nutritional depletion. CONCLUSIONS: Depressive symptoms as well as fat and lean body composition, due to preserved BMI in those with nutritional depletion, must be investigated. PMID- 21636138 TI - Stereological analysis of age-related changes of testicular peritubular cells in men. AB - This work aimed to analyze quantitative changes of peritubular cells in testes of aged men. Testicular tissues were obtained from 42 aged men with advanced prostate cancer and 16 young men with biopsy, quantitatively investigated with stereological techniques with quadrate mask grid, measured the parameters volume density (V(V)), numerical density on area (N(A)), and numerical density (N(V)) with grid test points. No significant differences were found in cell ratio, peritubular cell number per tubule, diameter of seminiferous tubules between young and old men (p>0.05). Aged men had higher pathologic assignment score than that of young men, which demonstrated more severe pathologic changes (p<0.05). Peritubular cell V(V) and pachytene germ cell V(V) increased significantly in old men compared to young men (p<0.05). Sertoli cell (SC) number per tubule in two dimensional was significantly less in aged men than that of young men, p<0.01. Peritubular cell N(A), N(V) decreased significantly in aged men compared to young one, p<0.05. It is concluded that the stereological data of peritubular cells from three-dimensional level in testes of aged men suggest a significant decrease when compared with young men, indicating age-related changes. PMID- 21636139 TI - Antigen carbohydrate 125 in heart failure: a promising clinical tool. PMID- 21636140 TI - Is four fatal for the Chinese? PMID- 21636141 TI - Short-term effect of pitavastatin on the reactive hyperemic index in post menopausal women with high levels in serum LDL-cholesterol. PMID- 21636142 TI - Mast cell activation disorders presenting with cerebral vasospasm-related symptoms: a "Kounis-like" syndrome? PMID- 21636143 TI - Catheter-based radiofrequency ablation therapy of the renal sympathetic-nerve system for drug resistant hypertension in a patient with end-stage renal disease. PMID- 21636144 TI - A clinical risk score for heart failure in patients with type 2 diabetes and macrovascular disease: an analysis of the PROactive study. AB - BACKGROUND: Diabetes mellitus is associated with a high risk for heart failure, which is further increased in the presence of coronary heart disease. So far no clinical risk score for development of heart failure exists for patients with type 2 diabetes and macrovascular disease. METHODS: Independent clinical predictors for heart failure events were identified by Cox regression in a post hoc analysis of the PROactive trial and were used for calculating a risk prediction score. RESULTS: 233 of 4951 patients with available baseline data suffered a serious adverse heart failure event during a mean follow-up of 34.5 (+/-2.3) months. Age, renal dysfunction, diuretic use, HbA1c, duration of diabetes, LDL-cholesterol, heart rate, right and left bundle branch block, microalbuminuria, previous myocardial infarction and pioglitazone treatment were independent predictors of heart failure. The risk score showed a good calibration and moderate discrimination (AUC 0.75). Patients were accurately stratified with an actual risk of 1.0%, 3.2% and 9.7% in the bottom, middle and top tertile of the score, respectively, with corresponding hazard ratios of 3.5 (95% CI 2.0-6.2) and 10.5 (95% CI 6.3-17.6) for the middle and top tertile compared to the bottom tertile (both p<0.0001). The score stratified well in subgroups defined by pioglitazone treatment, prior myocardial infarction, obesity, poor glycemic control and microalbuminuria. CONCLUSION: A risk score based on routinely assessed clinical variables proved a good stratification for future heart failure events in diabetic patients with macrovascular disease. Strategies targeting specific interventions and monitoring of high risk patients need further evaluation. PMID- 21636145 TI - 17beta-Estradiol enhances the recruitment of bone marrow-derived endothelial progenitor cells into infarcted myocardium by inducing CXCR4 expression. AB - BACKGROUND: 17beta-Estradiol (E2) has been thought to produce cardioprotective effects by mediating bone marrow-derived endothelial progenitor cells (EPC) for cardiac repair in the setting of acute myocardial infarction (AMI). However, the underlying mechanism of action of E2 on EPC remains unclear. CXCR4 is a critical modulator in homing of EPC. Accordingly, we hypothesized that E2 exerts beneficial effects through enhancing EPC homing to infarcted myocardium via mediating CXCR4 pathway. METHODS AND RESULTS: Migratory capacity and CXCR4 expression of EPC from ovariectomized BALB/C mice were detected after being incubated with various E2 concentrations for various incubation times. For in vivo studies, EPC were labeled with superparamagnetic ion oxide (SPIO) for tracing, and ovariectomized mice were grouped (n=11) after inducing AMI to receive saline without cells or with 3 * 10(6) non-preconditioned EPC, 100 nmol/L E2 preconditioned EPC, CXCR4 inhibitor AMD3100 (5 MUg/mL) preconditioned EPC, or EPC pretreated with E2 plus AMD3100. The number of homing EPC in infarcted myocardium and left ventricular (LV) function, dimensions and fibrosis were measured. In vitro data showed that E2 increased migratory activity and functional CXCR4 expression of EPC. However, these effects were completely blocked by AMD3100. In vivo data in E2 group displayed a greater number of homing EPC, decreased fibrosis of LV, and significant improvement in cardiac function. Nevertheless, effects of E2 preconditioning were abrogated by AMD3100. CONCLUSIONS: We conclude that E2 enhances the recruitment of EPC into infarcted myocardium by up-regulating functional CXCR4 expression, resulting in improving recovery after myocardial infarction. PMID- 21636146 TI - A meta-analysis of randomized trials of triple versus dual antiplatelet therapy after stent-based percutaneous coronary intervention. PMID- 21636147 TI - Radiation skin injury caused by percutaneous coronary intervention, report of 3 cases. PMID- 21636148 TI - Exercise as a beneficial adjunct therapy during Doxorubicin treatment--role of mitochondria in cardioprotection. AB - One of the mostly used chemotherapeutic drugs is the highly effective anthracycline Doxorubicin. However, its clinical use is limited by the dose related and cumulative cardiotoxicity and consequent dysfunction. It has been proposed that the etiology of this toxicity is related to mitochondrial dysfunction. The present review aimed to analyze the promising results regarding the effect of several types of physical exercise in cardiac tolerance of animals treated with acute and sub-chronic doses of Doxorubicin (DOX), highlighting the importance of cardiac mitochondrial-related mechanisms in the process. Physical exercise positively modulates some important cardiac defense systems to antagonize the toxic effects caused by DOX treatment, including antioxidant capacity, the overexpression of heat shock proteins and other anti-apoptotic proteins. An important role in this protective phenotype afforded by exercise should be attributed to mitochondrial plasticity, as related adaptations could be translated into improved cardiac function in the setting of the DOX cardiomyopathy. PMID- 21636149 TI - Quality of life improvement at midterm follow-up after transcatheter aortic valve implantation. AB - BACKGROUND: Transcatheter aortic valve implantation (TAVI) techniques have been presenting good procedural success and favorable clinical outcomes. However, optimal management of aortic valve disease in elderly patients depends on quality of life (QoL) improvement. In this study we aimed to evaluate changes in QoL in patients referred for TAVI. METHODS AND RESULTS: Prospective analysis of 74 consecutive patients (34 male), aged 81.6 +/- 8 years with symptomatic severe aortic valve stenosis (AS) ineligible for conventional aortic valve replacement, referred to TAVI in one tertiary center. For the assessment of QoL, the Minnesota Living with Heart Failure Questionnaire (MLHFQ) was used before the procedure and at 6.5 months. The mortality was 9.5% at 30 days and 20.2% at 6.5 months follow up. Fifty three (71.6%) patients completed MLHFQ at baseline and at follow-up. All patients showed good hemodynamic results and no signs of prosthesis dysfunction were observed on transthoracic echocardiography. The New York Heart Association (NYHA) class (2.9 +/- 0.4 to 1.4 +/- 0.7; p<0.001), and the MLHFQ scores [overall (37.0 +/- 14.7 vs. 14.4 +/- 10.1; p<0.001), physical dimension (23.2 +/- 9.5 vs. 8.6 +/- 5.9; p<0.001) and emotional dimension (5.4 +/- 4.2 vs. 2.6 +/- 3.0; p<0.001)] were significantly improved 6.5 months after TAVI. Patients with peripheral vascular disease (PVD) had an inferior improvement in QoL caused by a lower enhancement in physical dimension MLHFQ score (mean difference: -17.0 +/- 10.2 vs. -10.1 +/- 11.5; p=0.036). CONCLUSION: TAVI significantly improves symptoms and QoL in patients with severe AS and high surgical risk. Patients with PVD might be expected to have a less impressive improvement in QoL after TAVI. PMID- 21636150 TI - Authors' reply (Should tissue Doppler echocardiography be used to improve prediction of mortality risk in patients with Chagas cardiomyopathy?). PMID- 21636151 TI - Association of cardiorespiratory fitness with characteristics of coronary plaque: assessment using integrated backscatter intravascular ultrasound and optical coherence tomography. AB - BACKGROUND: Cardiorespiratory fitness (CRF) can predict future cardiovascular disease. Rupture of vulnerable plaque which often has a large lipid core with a thin fibrous cap causes acute coronary syndrome including sudden cardiac death. We tested our hypothesis that preserved CRF is associated with low lipid composition and thick fibrous cap thickness of coronary lesions. METHODS: We prospectively performed both integrated backscatter intravascular ultrasound (IB IVUS) and optical coherence tomography (OCT) for 77 non-culprit coronary lesions in 77 consecutive angina pectoris patients who underwent percutaneous coronary intervention (PCI). Percentage of achieved of predicted peak oxygen consumption (%PPeak Vo(2)) calculated based on measured peak Vo(2) using a cardiopulmonary exercise test performed post PCI was adapted as an indicator of patient CRF. RESULTS: Patients were divided into two groups [those with preserved CRF (%PPeak Vo(2) >82%) (Group I) or others (Group II)]. Coronary plaques of Group I patients had significantly smaller lipid volume, greater fibrous volume, and thicker fibrous cap thickness than those of Group II (32 +/- 14% vs. 45 +/- 13%, p<0.001; 57 +/- 11% vs. 49 +/- 11%, p<0.001; and 177.7 +/- 20.9 MUm vs. 143.7 +/- 36.9 MUm, p<0.001). In multivariate linear regression analysis, %PPeak Vo(2) showed a significantly negative correlation with lipid volume and a positive correlation with fibrous volume and fibrous cap thickness (beta=-0.418, p=0.001; beta=0.361, p=0.006; and beta=0.339, p=0.008). CONCLUSIONS: High %PPeak Vo(2) was associated with low lipid volume, high fibrous volume and thick fibrous cap thickness in coronary lesions. These results may well suggest an attenuated risk of cardiovascular events in patients with preserved CRF. PMID- 21636152 TI - Surviving acute myocardial infarction (IMA): a match ruled by insulin and mitochondria? PMID- 21636153 TI - A meta-analysis of randomized trials and adjusted observational studies of drug eluting stents versus coronary artery bypass grafting for unprotected left main coronary artery disease. PMID- 21636154 TI - Effect of statin therapy on contrast-induced nephropathy after coronary angiography: a meta-analysis. AB - BACKGROUND: Although the pleiotropic effects of statins are postulated to be renoprotective, clinical studies have demonstrated conflicting results. We undertook a meta-analysis of published trials to evaluate the impact of statin therapy on the incidence of contrast-induced nephropathy (CIN) in patients undergoing coronary angiography. METHODS: We searched MEDLINE and EMBASE databases through December 2010 for articles evaluating the effect of statins on the incidence of CIN in patients undergoing coronary angiography. Odds ratios (OR) with 95% confidence intervals (CI) were calculated using random effects modeling. RESULTS: Three randomized controlled trials involving 770 patients (330 in the statin group and 340 in the control group) and 7 non-randomized studies involving 31,959 patients (11,936 statin-pretreated and 20,023 statin-naive). The definition of CIN varied somewhat among the studies. Based on the pooled estimate across the 3 randomized controlled trials, statin therapy did not significantly reduce the incidence of CIN compared to control (OR=0.76, 95% CI: 0.41-1.41, p=0.39). No significant heterogeneity was found in the randomized studies (I(2)=0%, p=0.48). The pooled analysis of the non-randomized studies showed a marginally significant benefit associated with statin therapy (OR=0.60, 95% CI: 0.36-1.00, p=0.05). There was significant heterogeneity among the non-randomized studies (I(2)=88%, p<0.00001). CONCLUSIONS: Our meta-analysis suggests that statin therapy might be associated with a significant reduction in the incidence of CIN in patients undergoing coronary angiography. Further studies are warranted to clarify this issue. PMID- 21636155 TI - Volatile sulphur compounds-forming abilities of lactic acid bacteria: C-S lyase activities. AB - Volatile sulphur compounds (VSCs) are of prime importance in the overall aroma of cheese and make a significant contribution to their typical flavours. Thus, the control of VSCs formation offers considerable potential for industrial applications. Here, lactic acid bacteria (LAB) from different ecological origins were screened for their abilities to produce VSCs from L-methionine. From the data presented, VSC-forming abilities were shown to be strain-specific and were correlated with the C-S lyase enzymatic activities determined using different approaches. High VSCs formation were detected for those strains that were also shown to possess high thiol-producing abilities (determined either by agar plate or spectrophotometry assays). Moreover, differences in C-S lyase activities were shown to correspond with the enzymatic potential of the strains as determined by in situ gel visualization. Therefore, the assessment of the C-S lyase enzymatic potential, by means of either of these techniques, could be used as a valuable approach for the selection of LAB strains with high VSC-producing abilities thus, representing an effective way to enhance cheese sulphur aroma compounds synthesis. In this regard, this study highlights the flavour forming potential of the Streptococcus thermophilus STY-31, that therefore could be used as a starter culture in cheese manufacture. Furthermore, although C-S lyases are involved in both biosynthetic and catabolic pathways, an association between methionine and cysteine auxotrophy of the selected strains and their VSCs-producing abilities could not be found. PMID- 21636156 TI - DPP-IV inhibitor may affect spermatogenesis. AB - A 39-year-old physician was diagnosed retrograde ejaculation due to diabetes and treated with amoxapine. This treatment was effective; he could ejaculate. He began receiving DPP-IV inhibitor (Sitagliptin 50 mg, daily), in lieu of insulin injection. Unusual effect on semen quality was occurred following the administration of the drug. PMID- 21636157 TI - The role of acculturation in diabetes self-management among Chinese Americans with type 2 diabetes. AB - AIMS: We sought to examine the acculturation level and its relationship with diabetes self-management among Chinese Americans with type 2 diabetes. METHODS: We conducted a cross-sectional survey research study in a convenience sample of 211 Chinese Americans with type 2 diabetes. We measured acculturation and diabetes self-management and evaluated the association between acculturation and self-management behaviors after controlling participants' characteristics using structural equation modeling. RESULTS: The majority of the participants was born outside the U.S. and had a low level of acculturation. Women and older individuals were less acculturated, and those who had higher social economic status and lived in the U.S. for a longer period were more acculturated. The results of the structural equation modeling demonstrated that acculturation was significantly associated with DM self-management, and more-acculturated individuals were more likely to perform DM self-management than less-acculturated ones after controlling demographic characteristics. CONCLUSIONS: The results of this study indicated that Chinese Americans diagnosed with diabetes may benefit from acculturation to mainstream society probably because increased acculturation was associated with increased help seeking behaviors and increased use of professional services. PMID- 21636158 TI - Easy and flexible carbohydrate counting sliding scale reduces blood glucose of hospitalized diabetic patient in safety. AB - INTRODUCTION: The in-hospital sliding scale (Sc) to determine the insulin dose was changed to a carbohydrate counting sliding scale (CSc). Blood glucose levels before and after the change were compared. METHODS: The Sc was used in 32 patients in July and August 2009 (Sc group) and the CSc was used in 32 patients in September and October 2009 (CSc group). The blood glucose levels recorded before breakfast, lunch, and supper for 14 days were analyzed. The overall and daily mean of all blood glucose data were compared between the 2 groups. RESULTS: The overall blood glucose level was significantly lower in the CSc group than in the Sc group (p<0.001). The percentage of blood glucose level below 199mg/dL was 47% in the Sc group and 59% in the CSc group. The daily blood glucose level in the Sc group was 203-229mg/dL until day 14, while the daily mean blood glucose level decreased significantly to 186mg/dL on day 4 in the CSc group and remained in the 176-200mg/dL range on subsequent days (p=0.049). CONCLUSIONS: The CSc is easy to use, safe and useful in controlling the blood glucose level. PMID- 21636159 TI - Effects of 12-week overground walking training at ventilatory threshold velocity in type 2 diabetic women. AB - This study analyzed the effects of overground walking training at ventilatory threshold (VT) velocity on glycaemic control, body composition, physical fitness and lipid profile in DM2 women. Nineteen sedentary patients were randomly assigned to a control group (CG; n=10, 55.9+/-2.2 years) or a trained group (TG; n=9, 53.4+/-2.3 years). Both groups were subjected to anthropometric measures, a 12-h fasting blood sampling and a graded treadmill exercise test at baseline and after a 12-week period, during which TG followed a training program involving overground walking at VT velocity for 20-60min/session three times/week. Significant group*time interactions (P<0.05) in glycated hemoglobin (HbA1c), body mass, body mass index (BMI), peak oxygen uptake (VO(2peak)) and exercise duration were observed as effects of training exercise, whereas intervention did not induced significant changes (P>0.05) in fasting blood glucose, submaximal fitness parameters and lipid profile. Our results suggest that overground walking training at VT velocity improves long term glycaemic control, body composition and exercise capacity, attesting for the relevance of this parameter as an effective strategy for the exercise intensity prescription in DM2 population. PMID- 21636160 TI - Exercise capacity in relation to body fat distribution and muscle fibre distribution in elderly male subjects with impaired glucose tolerance, type 2 diabetes and matched controls. AB - BACKGROUND: The aim of this study was to examine the impact of insulin sensitivity and muscle fibre composition to exercise capacity in individuals with type 2 diabetes (T2D), impaired glucose tolerance (IGT) and normal glucose tolerance (NGT). METHODS: Thirty-nine male patients with T2D, 44 male subjects with IGT and 58 subjects with NGT matched for age, weight and body mass index (BMI) participated in the study. Insulin sensitivity was obtained with hyperinsulinemic-euglycemic clamps, muscle fibre distribution with a biopsy and exercise capacity from an incremental exercise test. Anthropometric measurements as height, weight, waist and hip circumference were performed. RESULTS: There were small differences between groups in waist hip ratio (WHR) with significance attained between NGT and T2D. There was a progressive reduction in exercise capacity, both expressed as VO(2peak) and work rate from subjects with NGT to IGT to T2D. Multiple regression analysis with VO(2peak) as dependent variable showed insulin sensitivity to be the most important factor followed by Type I fibres. WHR and capillary density also influenced the variance of VO(2peak). CONCLUSION: Exercise capacity is independently related to insulin sensitivity, muscle fibre composition and WHR in subjects with NGT, IGT and T2D who are matched for age and BMI. PMID- 21636161 TI - Response at 3 months to insulin dose decisions made at exenatide initiation in the Association of British Clinical Diabetologists (ABCD) nationwide exenatide audit. AB - It is uncertain what should be done with insulin dose if starting exenatide. In the ABCD nationwide exenatide audit, many patients with type 2 diabetes had worsened glycaemia when insulin was stopped. If starting exenatide, insulin should not be stopped but weaned off only if there is significant glycaemic response. PMID- 21636162 TI - Uric acid and mortality from all-causes and cardiovascular disease among adults with and without diagnosed diabetes: findings from the National Health and Nutrition Examination Survey III Linked Mortality Study. AB - Using data from the National Health and Nutrition Examination Survey III Linked Mortality Study, uric acid concentration was significantly related to mortality from all-causes (978 diabetic participants: hazard ratio per mg/dl, 1.14; 95% confidence interval, 1.01-1.28; 12,824 nondiabetic participants: hazard ratio, 1.06; 95% confidence interval, 1.02-1.11) but not major CVD. PMID- 21636163 TI - Psychosocial problems in patients with newly diagnosed diabetes: number and characteristics. AB - Early in the course of diabetes, it is important to identify and support patients whose psychosocial situations and reactions to the diagnosis may affect their ability to adjust or take adequate responsibility for self-care. We aimed to identify (a) the number and characteristics of patients, 18-65 years, newly diagnosed with diabetes, who needed psychosocial interventions and (b) the type of psychosocial problems they had. A total of 106 patients (72 men) were included in the study. Interviews showed that 41.5% had psychosocial problems. Fifteen dropped out early in the study; 38% of those remaining had psychosocial problems (PSP). More than half had problems with their life situation; most commonly in relationships. About a third had problems related to diabetes, most commonly, work-related. Compared to other participants, PSP patients lived in more strained social situations, especially regarding personal finances and social support. More of the PSP patients were anxious and depressed. They used negative coping strategies more often and more frequently expected that diabetes would negatively affect their future. In conclusion, early in the course of diabetes, screening instruments should be used to identify PSP patients. Treatment by medical social workers skilled in diabetes care should be offered. PMID- 21636164 TI - Privacy-preserving models for comparing survival curves using the logrank test. AB - The incorporation of electronic health care in medical institutions will benefit and thus further boost the collaborations in medical research among clinics and research institutions. However, privacy regulations and security concerns make such collaborations very restricted. In this paper, we propose privacy preserving models for survival curves comparison based on logrank test, in order to perform better survival analysis through the collaboration of multiple medical institutions and protect the data privacy. We distinguish two collaboration scenarios and for each scenario we present a privacy preserving model for logrank test. We conduct experiments on the real medical data to evaluate the effectiveness of our proposed models. PMID- 21636165 TI - [Acute isolated hydrops associated with atypical corneal condition in the fellow eye]. PMID- 21636166 TI - Two-step algorithm for the detection of Clostridium difficile from stool samples. PMID- 21636167 TI - Reduction in antibiotic use through procalcitonin testing in patients in the medical admission unit or intensive care unit with suspicion of infection. AB - We report an evaluation of the utility of serum procalcitonin (PCT) measurement as an additional diagnostic tool to support initiating or withholding antibiotics in clinical situations where there is a clinical suspicion of infection but the diagnosis is uncertain. During a six-month period, 99 patients on the medical admission unit (MAU) with suspected infection, and 42 patients on the intensive care unit (ICU) with clinical signs or physiological parameters suggesting possible new infection, had serum PCT concentration measured with the result available within 90min of the request. The test was initiated by the microbiology/infection team during clinical consultations to support the antibiotic decision. On the basis of low PCT values, antibiotics were withheld in MAU on 52 occasions and in ICU on 42 occasions. Patients were followed up prospectively for a week. There was neither progression of bacterial infection requiring antibiotics, nor complications or infection-related mortality in any patients who were denied antibiotics on either MAU or ICU. Without the PCT value it is likely that all of these patients would have received empirical antibiotics. Reduction in unnecessary antibiotic usage was made without any adverse effects on these patients and there was a clear reduction in antibiotic prescribing with cost reduction implications. PCT has the potential to become a valuable tool in antibiotic management. PMID- 21636168 TI - Glove removal method and distance: what else can affect contamination? PMID- 21636169 TI - Occupational exposure to blood in workers in a Greek hospital. PMID- 21636170 TI - Effectiveness of a hand hygiene promotion strategy using alcohol-based handrub in 6 intensive care units in Colombia. AB - BACKGROUND: Hand hygiene is an effective strategy for the prevention of health care-associated infection (HAI). We investigated the effect of a hand hygiene promotion strategy introducing alcohol-based handrub (AHBR) on the incidence of HAI in a university hospital in Colombia. METHODS: A Prospective cohort study was performed in 6 intensive care units from January 2001 to December 2005. HAI were identified using standard US Centers for Disease Control and Prevention definitions. Alcohol-based handrub dispensers were installed between February and June 2002. RESULTS: Total ABHR consumption was 5,794 L (mean, 28.9 L per 1,000 patient-days) and significantly increased over time (+9.2% per year; P < .001). Of 14,516 patients cumulating 166,498 patient-days, 2,398 (16.5%) acquired a total of 3,490 HAI episodes (20.9 per 1,000 patient-days). Incidence densities for central line-associated bloodstream infection (CLABSI), ventilator-associated pneumonia, and urinary tract infections were 7.7, 10.6, and 3.6 episodes per 1,000 device-days, respectively. A significant decrease was observed for CLABSI ( 12.7% per year; P < .001) with low nurse-to-patient ratio independently associated with infection (odds ratio, 1.11; 95% confidence interval: 1.07-1.16; P < .001). CONCLUSION: Improved hand hygiene measured by increased ABHR consumption resulted in CLABSI reduction. Low nurse-to-patient ratio is independently associated with HAI in an upper-middle income country. PMID- 21636171 TI - Before influenza tests results are available, can droplet precautions be instituted if influenza is suggested by leukopenia, relative lymphopenia, or thrombocytopenia? PMID- 21636172 TI - Benchmarking of percutaneous injuries at a teaching tertiary care center in Saudi Arabia relative to United States hospitals participating in the Exposure Prevention Information Network. AB - BACKGROUND: Despite recent improvements in policies, practices, and device design, percutaneous injuries (PIs) from needles and sharp instruments continue to expose health care workers to the risk of bloodborne pathogens. METHODS: Prospective surveillance was instituted to study the epidemiologic characteristics of PIs at King Abdulaziz Medical City, Riyadh, Saudi Arabia (KAMC R) from 2004 through 2008 and to benchmark these characteristics relative to those of a network of US hospitals participating in the Exposure Prevention Information Network (EPINet) research group (2004-2007). RESULTS: The mean PIs rate per 100 daily occupied beds in KAMC-R was significantly lower than that reported by teaching and nonteaching US EPINet hospitals. Similar to US EPINet hospitals, nurses at KAMC-R reported more PIs than physicians. Compared with US EPINet hospitals, nurses at KAMC-R experienced more PIs (52.8% vs 38.1%, respectively), whereas physicians experienced fewer PIs (18.4% vs 28.6%, respectively). The majority of PIs happened in patient wards (50.6%) in KAMC-R and operating rooms (34.1%) in US EPINet hospitals. Suturing, which was involved in only 10% of PIs at KAMC-R, was the most frequent PIs mechanism in US EPINet hospitals (23.3%). In both KAMC-R and US EPINet hospitals, disposable syringes were the most frequent devices involved, the left hand was the most common site of injury, and the source patient was largely identifiable. CONCLUSION: We have lower rates of PIs at KAMC-R relative to US EPINet hospitals. Whereas it is difficult to fully explain such differences, this could be due to variations in health care systems, underreporting, or the impact of the PIs prevention activities. PMID- 21636173 TI - [Pulmonary thromboembolism and congenital absence of the inferior vena cava]. PMID- 21636174 TI - [Granular cell tumor of the esophagus: description of an infrequent benign tumor]. AB - Granular cell tumors (GCT) are infrequent tumors first described by Abrikossoff in 1926. Gastrointestinal involvement occurs in about 6% of GCT, the esophagus being the most frequent location. These tumors are usually benign and asymptomatic and are usually found incidentally when an upper gastrointestinal endoscopy is carried out for another reason, showing an isolated polyp or sessile submucosal nodule, covered by intact yellowish mucosa and with firm consistency. Endoscopic ultrasonography has significantly improved the diagnosis of these lesions. Nowadays endoscopic mucosectomy is the treatment of choice of esophageal GCT with a low frequency of complications. Histologic analysis of the surgical specimen shows specific characteristics such as positivity for S-100 protein. We present two new cases of esophageal GCT that were diagnosed recently and discuss the most relevant features of this infrequent disease. PMID- 21636175 TI - [Corticosteroid-induced mediastinal lipomatosis in a patient with overlap syndrome (autoimmune hepatitis-primary biliary cirrhosis)]. PMID- 21636176 TI - [Care of the chronic patient in a complex situation: the challenge of building an integrated care scenario]. AB - The ageing of the population and the increase in people with chronic illnesses is a scenario that creates important challenges on how to organise and provide services for this population group. There is already experience and evidence that would help us to re-think about what are the key aspects that should be taken into account to design a simple health model directed towards this new paradigm.The clinical care approach in itself is important, but at the same time, limited. A multidimensional approach is required where different elements, such as real platforms for change, are incorporated; the contribution that can be made by information systems, the objectives that should be assigned cross-sectionally to different organisations and professionals, the clinical and relational skills that should be contemplated in training and skill development plans or how the provision should be financed. Also, only interacting with some of these elements could give rise to a significant change in the care of chronic patients, particularly those in a highly complex and vulnerable situation. PMID- 21636177 TI - [Validity and usefulness of B-type natriuretic peptide (BNP) for early detection of left ventricular dysfunction in high-risk patients in primary care]. AB - OBJECTIVE: The aim of this study was to determine the accuracy of BNP test for early diagnosis of left ventricular dysfunction in patients at high-risk for heart failure. DESIGN: Cross-sectional descriptive study. SETTING: 7 Primary Care Centres in Madrid (Spain). PARTICIPANTS: A consecutive sample of 204 consecutive asymptomatic patients with high risk for heart failure (Stages A-B, AHA/ACC Classification). MAIN MEASUREMENTS: BNP plasma levels were measured in the clinical setting using Triage BNP Test((r)) (Biosite((r))) and an echocardiography was performed in the following 3 days in a single hospital unit as a reference standard. Plasma BNP levels were compared depending on the presence/absence of left ventricular dysfunction (LVD), type and severity degree. Sensitivity, specificity, positive and negative predictive values, and Area under the receiver operating characteristic curve (ROC) for BNP assay were calculated. RESULTS: BNP values were significantly higher (P<.001) in patients with left ventricular systolic dysfunction (LVSD). No significant differences were found for diastolic dysfunction. The best cut-off value to discriminate the patients with LVSD was 71.00 pg/ml, with an Area under the ROC curve of 0.757 (95% CI 0.64 0.87). Sensitivity for LVD diagnosis was 75% (95% CI 50.66-99.34), specificity 70.19% (95% CI 62.81-77.57), positive predictive value (PPV) 20% (95% CI 9.05 30.95), and negative predictive value (NPV) 96.58% (95% CI 92.86-100), with LVSD prevalence of 9.04% in this population. CONCLUSIONS: BNP determinations are of value in diagnosing LVSD in a primary care setting, with similar sensitivities and specificities. Due to the high NPV is useful to rule-out patients for echocardiography. PMID- 21636178 TI - [Use of a proxy to the Charlson index to study the short and long-term comorbidity and mortality in the elderly]. AB - OBJECTIVES: To design a proxy to the Charlson comorbidity index (PrCCI) for use in studies with pre-existing data sources and to analyse its influence on mortality risk in elderly people. DESIGN: Longitudinal study. SETTING: Population base, From "Aging in Leganes" cohort. PARTICIPANTS: All individuals with complete information (n=978, aged 65 years old and older) were included in the construction of the proxy (PrCCI). MEASUREMENTS: A proxy (PrCCI) was created based on the original Charlson Comorbidity Index (CCI) and the available pre existing data from the "Aging in Leganes" study. The relationship between PrCCI and mortality was assessed using a Survival analysis. Cox proportional Hazard Models were adjusted for possible confusion factors. RESULTS: The PrCCI varied from 0 to 11 points (median=2 points, RI25-75=1-3). It was higher in women than in men until 79 years old, but no differences were found for both men and women from 80 years old. Individuals who scored four and more points in the new index had a higher mortality risk after 5 and 15 years of follow up, even after controlling for sociodemographic, health behaviours and health status related covariables (HR: 3.69, 95% CI: 1.52-8.96 and 2.14, 95% CI: 1.42-3.21, respectively). CONCLUSION: The proxy to the Charlson comorbidity index is easy and useful to measure comorbidity in studies in population from 65 to 75 years old, which use pre-existing data bases and data available in the primary care setting. The association between PrCCI and mortality shows that it is useful as comorbidity index. PMID- 21636179 TI - [Meningoencephalitis due to varicella-zoster virus: an uncommon problem of acute confusional syndrome in the elderly]. PMID- 21636180 TI - Design, synthesis and antiproliferative activities of biarylolefins based on polyhydroxylated and carbohydrate scaffolds. AB - A series of diversely substituted biarylolefins based on carbohydrate and dihydroxyethylene scaffolds were synthesized and evaluated for antiproliferative activity against a panel of human tumor cell lines. Among the thirty-five yet unknown biarylolefins prepared, six displayed potent antiproliferative activities with IC(50) values in the micromolar and submicromolar range. As a new type of antiproliferative agent, the most potent compound 26 showed an IC(50) value of 70 nM against SK-OV3 cell line (ovarian cancer). All the synthesized compounds exhibited a poor or modest tubulin polymerization inhibitory activity suggesting another mode of action for these compounds. Molecular docking simulations to the colchicine binding site of tubulin of representative compounds have been used to explain the lack of activity as inhibitors of tubulin polymerization. PMID- 21636181 TI - A review of clinical competence assessment in nursing. AB - AIMS: To investigate trends in the evaluation of clinical competence in nursing students and newly qualified nurses over the last 10 years. DESIGN: A literature review following PRISMA guidelines. METHODS: The following databases were searched: Cochrane, Medline and CINAHL using the terms competenc*, nurs*and assess*, evaluat*, measure*, from January 2001-March 2010. Strict inclusion and exclusion criteria were applied. RESULTS: Twenty three papers were included and these mainly considered the following topics: instrument development and testing; approaches to testing competence; assessment and related factors. A holistic concept of competence is gaining popularity, and consensus around definitions is emerging. Some methods and instruments to measure competence are under systematic development and testing for reliability and validity with large samples and rigorous statistical method. Wider national and international cooperation is evident in competence-based assessment. CONCLUSIONS: Competence-based education is evident, but this does not mean that issues related to competence definition have been resolved. Larger and more international cooperation is required to reach common agreement and validity in competence-based education and assessment. PMID- 21636182 TI - Student nurses' attitudes to illicit drugs: a grounded theory study. AB - The aim of this research was to identify the factors that influence the attitudes of student nurses towards illicit drugs. This insight is important in providing a foundation for the development of educational approaches aimed at challenging what appear to be negative attitudes to illicit drug users within nursing. The absence of a testable hypothesis prior to the investigation led to the generation of theory from the data (inductive enquiry) with a constructivist approach to grounded theory (Charmaz, 2006), being employed. Data generation involved informal conversational interviews (n=12), semi-structured interviews (n=9), four focus groups and an audit of the education received by students (n=61) around substance misuse issues. The final grounded theory indicated that: Student nurses enter training with a wide range of personal experiences relating to illicit drug use. The influences of society's negative views and the image of drug use presented in the press appeared to be significant factors in developing their attitudes on the subject. In the absence of effective approaches to education, and given that many professionals in the practice environment appear to view illicit substance users in a negative way, it is likely that interventions with identified drug users will be influenced by negative attitudes. PMID- 21636183 TI - Effect of essential amino acid supplementation on quality of life, amino acid profile and strength in institutionalized elderly patients. AB - BACKGROUND & AIMS: This study assessed the efficacy of supplemented essential amino acids on depressive symptoms, nutrition, muscle function, daily physical activity, and health-related quality of life (HRQoL) of institutionalized elderly patients. METHODS: Forty-one patients (58.5% women; mean age 79.8 yrs) with sequelae of coronary artery disease (73%), femoral fracture (34%), were randomly assigned to receive oral essential amino acids 4 gr 2 times a day for 8 weeks or isocaloric placebo. Before randomization and 8 weeks after the protocol started, the following variables were measured: depressive symptoms (Geriatric Depression Scale, GDS), nutritional panel (Mini Nutritional Assessment, MNA; serum albumin and prealbumin levels), muscle strength (Hand Grip, HG), Activity Daily Life (ADL), Quality of Life (SF-36, HRQoL) and amino acid profile. RESULTS: Compared with the placebo group, EAA patients improved nutrition (MNA score 22.6 +/- 1.5 post vs 21.8 +/- 1.6 pre; p < 0. 04, albumin g/dl 4.04 +/- 0.35 post vs 3.88 +/- 0.3 pre; p < 0.01), GDS(score 10.3 +/- 1.75 post vs 13.85 +/- 3.37 pre; p < 0.001), HG (Kg 19.75 +/- 1.7 post vs 18.68 +/- 1.36 pre; p = 0.001), ADL (p < 0.04) and both physical and mental components of SF-36 (p < 0.002). CONCLUSIONS: Oral supplementation with essential amino acids improved several determinants of quality of life in institutionalized elderly patients, including depressive symptoms, nutrition, muscle function and daily life activity. PMID- 21636184 TI - Investigating incorporation and distribution of radionuclides in trinitite. AB - Most of the surface explosions in nuclear tests have released radioactivity to the environment in the form of bulk glassy materials originating from the melting of sandy soil in the neighbourhood of ground zero. In view of clarifying issues concerning the mechanism of formation and the radiological impact of these materials, we investigated incorporation and volume distribution of radionuclides in a typical fragment of trinitite, the glassy substance generated following the first nuclear test (Trinity Site, New Mexico, 1945). Specific activities were determined by gamma-spectrometry for the most significant fission and activation products. In particular, (152)Eu activity was used to estimate the original point of collection of the sample with respect to ground zero. After embedding in an epoxy resin, the sample was then sliced to perform cross-sectional beta- and alpha-autoradiograph. alpha-spectrometry was also carried out on a fine powder obtained by surface abrasion. In the beta-autoradiography, hot spots were distinguishable in the proximity of the blast side, over a 1000 times less intense background of sand activation products. Also alpha-contamination (from (239+240)Pu and (241)Am) was mostly concentrated within the superficial layer, in a fraction of only 20% of the overall volume of the sample, exhibiting a discontinuous, droplet-like distribution. This evidence would partially support a recent hypothesis on trinitite formation according to which most of the glass layer was formed not on the ground but by a rain of material injected into the fireball that melted, fell back, and collected on a bed of already fused sand. PMID- 21636185 TI - Indoor radon levels in Greek schools. AB - Radon and gamma dose rate measurements were performed in 512 schools in 8 of the 13 regions of Greece. The distribution of radon concentration was well described by a lognormal distribution. Most (86%) of the radon concentrations were between 60 and 250 Bq m(-3) with a most probable value of 135 Bq m(-3). The arithmetic and geometric means of the radon concentration are 149 Bq m(-3) and 126 Bq m(-3) respectively. The maximum measured radon gas concentration was 958 Bq m(-3). As expected, no correlation between radon gas concentration and indoor gamma dose rate was observed. However, if only mean values for each region are considered, a linear correlation between radon gas concentration and gamma dose rate is apparent. Despite the fact that the results of radon concentration in schools cannot be applied directly for the estimation of radon concentration in homes, the results of the present survey indicate that it is desirable to perform an extended survey of indoor radon in homes for at least one region in Northern Greece. PMID- 21636186 TI - Direct and tomographic dimensional analysis of the inter-radicular distance and thickness of the vestibular cortical bone in the parasymphyseal region of adult human mandibles. AB - We calculated the inter-radicular distances between the canine, and first and second premolars, of human mandibles and the thickness of the cortical bone at adjacent sites using computed tomographic (CT) imaging, and assessed the precision of the dimensional assessment made by CT compared with the same measurement made directly with calipers. We examined 100 adult cadaveric dentate human hemimandibles. At the level at which monocortical screws are inserted to place miniplates according to the current technique used to treat mandibular fractures, points A, B, and C referred to the canine, and first and second premolars, and were marked to calculate the level of the CT slice and as the reference for sectioning of the hemimandible. Our findings showed that there was no significant difference in the inter-radicular distance or in the thickness of cortical bone between the sides of the mandible. The vestibular cortical bone was less than 3.0mm thick in 91 of the samples. In 98 of the samples the inter radicular distance between the canine and the first and second premolars was more than 2mm. There was no significant difference in micrometric precision between the dimensional assessment on CT and direct measurement using a caliper. In the region of the mental foramen the screws have cortical bone less than 3mm thick in which to be anchored. The inter-radicular distance suggests that there is a minimal risk of radicular injury when miniscrews are inserted between the alveolar structures. CT is a reliable tool for measuring the inter-radicular distance and the thickness of mandibular cortical bone. PMID- 21636187 TI - Analysis of the impact of deprivation on urgent suspected head and neck cancer referrals in the Mersey region between January 2004 to December 2006. AB - Serious delay in patients presenting with head and neck cancer is associated with poor outcomes. We aimed to examine the influence of deprivation on professional delay in the Mersey region from 2004 to 2006. The study sample comprised 6681 patients who were referred between January 2004 and December 2006. The dataset was dominated by the largest hospital (H1), which received 48% of all cases. Median referral overall was 12 days (IQR 8-15 days), and 74% of patients were referred in 14 days or less. Professional delay (percentage 14 days or less) was associated with hospital (from 58% H1 to 97% H5), year of referral (from 64% in 2004 to 80% in 2006), age (from 69% under 55 years to 80% over 75 years), and deprivation (Index of Multiple Deprivation 2000 from 67% most deprived (IMD 1) to 85% least deprived (IMD 5)). Hospital location was associated with these factors and the results imply that by far, the most important variable in predicting professional delay was the hospital that received the referral. Trends over time in age, and to a lesser extent, for deprivation were noted in H1, but were largely absent across other hospitals. Some of them needed to make substantial improvements to meet the two-week referral pathway and it would be interesting to compare these results with current practice. This study highlights the importance of maintaining the standards of the current policy on two-week referrals for suspected head and neck malignancy. PMID- 21636188 TI - A double-blind, placebo-controlled, randomised trial of active manuka honey and standard oral care for radiation-induced oral mucositis. AB - Our aim was to investigate the effect of active manuka honey on radiation-induced mucositis. A total of 131 patients diagnosed with head and neck cancer who were having radiotherapy to the oral cavity or oropharyngeal area were recruited into the study, and were randomly allocated to take either manuka honey or placebo (golden syrup) 20 ml 4 times daily for 6 weeks. Mucositis was assessed according to the Radiation Therapy Oncology Group (RTOG) scale at baseline, weekly during radiotherapy, and twice weekly thereafter until the mucositis resolved. The patient's weight was recorded at the same time as the mucositis was assessed. Throat swabs to identify bacterial or fungal infections were taken at baseline, and during and after radiotherapy. There was no significant difference between honey and golden syrup in their effects on mucositis. Active manuka honey did not improve mucositis, but both the honey and the syrup seemed to be associated with a reduction in bacterial infections. Compliance was a problem after the onset of mucositis, which may have affected the findings. PMID- 21636189 TI - The effect of sutureless wound closure on postoperative pain and swelling after impacted mandibular third molar surgery. AB - Our aim was to assess the influence of sutureless and multiple-suture closure of wounds on postoperative complications after extraction of bilateral, impacted, mandibular third molars in 30 patients in a split mouth study. After the teeth had been removed, on one side the flap was replaced but with no suture to hold it in place (study side), and on the other side the wound was closed primarily with three sutures (control side). Recorded complications included pain, swelling, bleeding, and formation of periodontal pockets. The results showed that patients had significantly less postoperative pain and swelling when no sutures were used (p=0.005). There were no signs of excessive bleeding or oozing postoperatively on either side. Six months postoperatively there was no significant difference in the depth of the periodontal pocket around the second molar. PMID- 21636190 TI - Biomechanical properties of the scapholunate ligament and the importance of its portions in the capitate intrusion injury. AB - BACKGROUND: Repair of the dorsal component of the scapholunate ligament alone is the usual surgical treatment for scapholunate injuries. Recent literature has suggested that additionally repairing the palmar component of the scapholunate ligament leads to improved and lasting clinical outcomes. The aim of this study was to determine the biomechanical properties of both portions of scapholunate ligaments derived from the same wrist and compare them with the whole scapholunate ligament. The goal was to further elucidate the importance of the palmar portion of the scapholunate ligament from a biomechanical perspective. METHODS: Scapholunate ligaments and their components were harvested from the same fresh frozen cadaveric wrists. Force at failure and stiffness were measured. FINDINGS: The mean maximum loads to failure for the entire scapholunate ligament, domicronrsal and palmar portions were found to be 147 (SD 54)N, 83 (SD 18)N and 86 (SD 16)N respectively. No statistical difference was found between the mean maximum load and stiffness for palmar and dorsal components (P=0.05). Mean maximum load and stiffness, of the intermediate portion, were 36 (SD 15)N and 25 (SD 23)N/mm. INTERPRETATION: Our biomechanical findings on the dorsal and palmar portions of the scapholunate ligament suggest that each portion contributes approximately 50% to the whole ligament tensile force. These results appear to agree with other reports about the stabilizing role of the palmar portion of the scapholunate ligament and suggest that the palmar portion of the ligament should be considered for surgical repair. PMID- 21636191 TI - The fate of the aqueous phase polycyclic aromatic hydrocarbon fraction in a detention pond system. AB - The concentration of dissolved polycyclic aromatic hydrocarbons (PAH) in influent, effluent, and within a detention pond system was measured. The "soluble fraction" was operationally defined as the PAHs in solution that passed through a 1.2 MUm filter. The results show that influent and effluent PAH concentrations were similar, indicating that dissolved PAH moved essentially unhindered through the detention pond system. In general, low molecular weight PAH were present at the highest concentrations and the highest PAH concentrations were measured in Summer. Also, year-to-year variations in PAH concentration were observed. At the end of sufficiently large storms, the pond was comparably unpolluted. During dry periods, the dissolved PAH concentration rose, possibly due to evapoconcentration and by partitioning of PAH from trapped contaminated sediment in the detention pond system. This study provides evidence that aqueous-phase PAH concentrations in runoff water were relatively unaffected by the passage through a conventional detention pond system. PMID- 21636192 TI - Comparison of competitive and synergetic adsorption of three phenolic compounds on river sediment. AB - Knowledge of toxic chemical sorption by soil/sediment is critical for environmental risk assessment of toxic chemicals, especially for the multi sorbate system in river ecosystem. Sorption characteristics of 2, 4 Dichlorophenol, 2, 4-Dinitrophenol and 2, 4-Dimethyphenol on sediment were investigated. Adsorption isotherms in single- and multi-sorbate systems fitted well the Freundlich model. The adsorption effects were different among three selected phenolic compounds in single- and multi-sorbate systems. The synergetic affect that 2, 4-Dinitrophenol and 2, 4-Dimethyphenol bring to 2, 4 Dichlorophenol can be explained by the compression of double electronic layer and the charge neutrality. Adsorption kinetic results showed that pseudo-second-order model can be used to describe the experimental data and the adsorption affinity of phenolic compounds influenced greatly by the adsorption velocity. The present study suggests that the fate and transport of emerging pollutants such as phenolic compounds could be affected in the presence of different hydrophobic pollutants in aquatic systems. PMID- 21636193 TI - Vitamin D status and mortality risk in CKD: a meta-analysis of prospective studies. AB - BACKGROUND: Vitamin D deficiency, assessed as low 25-hydroxyvitamin D (25[OH]D) level, is highly prevalent in patients with chronic kidney disease (CKD) and is associated with various adverse health outcomes. Whether low 25(OH)D levels in patients with CKD are an independent risk factor for mortality remains to be studied in detail, and this was the objective of our work. STUDY DESIGN: A systematic review and meta-analysis of prospective observational studies. SETTING & POPULATION: Patients with CKD. CKD was diagnosed mainly as decreased estimated glomerular filtration rate. SELECTION CRITERIA FOR STUDIES: We performed a systematic literature search in MEDLINE, ISI, and EMBASE to identify prospective studies reporting on 25(OH)D levels and mortality. PREDICTOR: 25(OH)D serum concentrations. OUTCOME: All-cause mortality. RESULTS: 10 studies with an overall sample of 6,853 patients with CKD were included. Relative risk of mortality per 10-ng/mL (25-nmol/L) increase in 25(OH)D level was 0.86 (95% CI, 0.82-0.91), with no indication of publication bias or significant heterogeneity (I(2) =15%; P = 0.3). Summary estimates for CKD cohorts with and without dialysis treatment showed homogeneous results (P = 0.9). LIMITATIONS: Results may be limited by heterogeneity, unconsidered confounders, and the observational design of the studies. Furthermore, publication bias by unpublished null findings on the association of 25(OH)D level and mortality cannot be ruled out and ascertainment of CKD was based largely on estimated glomerular filtration rate. CONCLUSIONS: Higher 25(OH)D levels are associated with significantly improved survival in patients with CKD. Whether treatment of low 25(OH)D level using natural vitamin D supplementation improves survival in patients with CKD remains to be elucidated in randomized controlled trials. PMID- 21636194 TI - Socioeconomic inequalities in old-age mortality: a comparison of Denmark and the USA. AB - Previous studies have reported important variations in the magnitude of health inequalities between countries that belong to different welfare systems. This suggests that there is scope for reducing health inequalities by means of country level interventions. The present study adds to this literature by exploring whether the magnitude of socioeconomic inequalities in mortality is associated with social inequality levels. Denmark and the USA belong to fundamentally different welfare systems (social democratic and liberal) and our study thereby contributes to the ongoing debate on whether welfare systems are linked to health inequalities. We analyze Denmark and the USA in terms of socioeconomic differences in mortality above age 58. The data sources were Danish register data from 1980 to 2002 (n = 2,029,324), and survey data from the US Health and Retirement Study (HRS) from 1992 to 2006 (n = 9374). Survival analysis was used to study the impact of socioeconomic status on mortality and the magnitude of mortality differences between the two countries was compared. The results showed surprisingly that mortality differentials were larger in Denmark than in the USA even after controlling for a number of covariates: The poorest 10 percent of the Danish elderly population have a mortality rate ratio of 3.32 (men) and 3.70 (women) compared to the richest 25 percent. In the USA the corresponding rate ratios are 1.67 and 1.56. Low income seems to be a more powerful risk factor for mortality than low education. A number of possible explanations for higher mortality differences in Denmark are discussed: unintended positive correlation between generous health services and health inequality, early life influences, mortality selection, and relative deprivation. PMID- 21636195 TI - Forging convictions: the effects of active participation in a clinical trial. AB - This qualitative study explored non-specific influences on participation in, and outcomes of, a randomised controlled trial. It was nested within a single-blind clinical trial of western acupuncture which compared real acupuncture with two types of placebo control administered to National Health Service (NHS) patients awaiting hip and knee replacement surgery in England. Data collection (2004-2008) was based on narrative-style interviews and participant observation. The results indicate that trial recruitment and retention depend on a set of convictions forged largely as a result of contextual factors peripheral to the intervention, including the friendliness and helpfulness of research centre staff and status of the administering practitioner. These convictions also influence the reporting of the study outcomes, particularly if participants experience uncertainties when choosing an appropriate response. The findings suggest that participants in clinical trials are actively involved in shaping the research process, rather than passive recipients of treatment. Thus the outcomes of trials, notably those involving contact interventions, should be regarded not as matters of fact, but as products of complex environmental, social, interpretive and biological processes. In this paper, we develop and present a 'theory of active research participation' which offers a framework for understanding the impact of non specific processes in clinical trials. PMID- 21636196 TI - Scouting for talent: appointment practices of women professors in academic medicine. AB - This paper contributes to current literature on the under-representation of women in academic medicine by critically examining appointment practices for medical professors in the Netherlands. By opening the black box of these highly secretive appointments, it is shown how allegedly gender-neutral practices contribute to the perpetuation of gender inequalities in academic medicine. The methods employed include quantitative analysis of 286 appointment reports and qualitative interviews with 21 scouts. The analysis revealed a dominant pattern of recruitment by invitation by male scouts, leading to three gender mechanisms of inclusion and exclusion through formal/informal networking. When candidates are recruited through homogeneous male networks, the pool of potential candidates is drastically restricted. Women are not seen as obvious choices for professorships since their commitment to the job is questioned. Furthermore, women do not correspond to the image of the ideal manager since they do not appear to conform to the gendered preconceptions of leadership held by the predominantly male scouts. PMID- 21636197 TI - Surgical treatment of giant fibrous dysplasia of the mandible with concomitant craniofacial involvement. PMID- 21636198 TI - [Inferior gluteal artery perforator flap: anatomical study for its application in breast reconstruction]. AB - PURPOSE: The inferior gluteal artery perforator flap, which is vascularised by perforator branches of the inferior gluteal artery (formerly ischiatic artery) is harvested in the gluteal crease. The purpose of this anatomical study was to clarify the anatomical features of this flap and locate the perforators of the inferior gluteal artery destined to the flap, in view to facilitate its surgical removal, as free flap for breast reconstruction. MATERIALS AND METHOD: We performed 12 dissections on fresh cadavers, after selective injection of the inferior gluteal artery with a gelified solution. During harvesting, we located the perforators which arise from the inferior gluteal artery and perfuse the flap. RESULTS: We distinguished four zones, defined relatively to anatomical landmarks of the region, according to the frequency of perforators. CONCLUSION: This anatomical study makes possible an improvement of the technique to raise this flap in its free shape with view to use it for breast autologus reconstruction especially given its numerous advantages: a constant volume of fat even in thin patients and minimal donor site morbidity. PMID- 21636200 TI - Therapeutic hypothermia after cardiac arrest - cerebral perfusion and metabolism during upper and lower threshold normocapnia. AB - BACKGROUND: During cardiac arrest and after successful resuscitation a continuum of ischaemia-reperfusion injury develops. Mild hypothermia exerts protective effects in the postresuscitation phase but also alters CO2 production and solubility, which may lead to deleterious effects if overlooked when adjusting the ventilation of the resuscitated patient. Using a multimodality approach, the effects of different carbaemic states on cerebral perfusion and metabolism were evaluated during therapeutic hypothermia. METHODS: Eight comatose survivors of prehospital cardiac arrest were cooled to 33 degrees C for 24 h and underwent a 60 min phase of interventional lower threshold normocapnia according to temperature non-corrected pCO2 (4.2 kPa) and higher threshold normocapnia according to corrected pCO2 (6.0 kPa) in a random order. Prior to, during and after each phase, cerebral perfusion and metabolites via a microdialysis catheter were measured. RESULTS: During upper-threshold pCO2, an increase in middle cerebral artery mean flow velocity (MFV) and jugular bulb oxygen saturation (jSvO2) were observed with a concomitant decrease in cerebral lactate concentration. Lower threshold normocapnia was associated with a decrease in MFV in most patients. In all patients jSvO2 decreased but no change in cerebral lactate was observed. In seven patients jSvO2 decreased below 55%. These changes were not reflected to intracranial pressure or cerebral oximetry. CONCLUSIONS: During induced hypothermia, lower threshold normocapnia was associated with decreased cerebral perfusion/oxygenation but not reflected to interstitial metabolites. Upper threshold pCO2 increased cerebral perfusion and reduced cerebral lactate. Vigilance over the ventilatory and CO2 analysis regimen is mandatory during mild hypothermia. PMID- 21636201 TI - Modulation of nitric oxide expression with methylene blue does not improve outcome after hypovolemic cardiac arrest. AB - AIM OF THE STUDY: We recently reported that female sex protects against cerebral and cardiac injury after hypovolemic cardiac arrest (CA), independent of sex hormone effects. As female sex was also associated with a smaller increase in inducible and neuronal nitric oxide synthase (NOS), we hypothesised that nitric oxide inhibition with methylene blue (MB) improves the outcome, primarily in male animals. METHODS: Twenty sexually immature piglets (10 males and 10 females) were bled to mean arterial blood pressure of 35mmHg, and were subjected to 2 min of untreated CA followed by 8 min of open chest cardiopulmonary resuscitation (CPR). Volume resuscitation was started during CPR with intravenous administration of 3mlkg(-1) hypertonic saline-dextran. Methylene blue was then administered as bolus of 2.5mgkg(-1) over 20 min, followed by 1.5mgkg(-1) infusion over 40 min. Historical data from 21 animals were used as control (no MB). Hemodynamic parameters, myocardial injury (troponin I), and short-term survival (3-h) were evaluated. Histopathological evaluation of heart specimens was performed. RESULTS: There were no differences between male and female animals in survival or resuscitation rate. After CA female piglets had significantly greater systolic and mean arterial pressures, and had lower troponin I plasma concentrations compared to male piglets, with or without MB. No difference was observed in histopathological analysis of heart specimens between sexes. CONCLUSIONS: After resuscitation from hypovolemic CA, female sex protects against cardiac injury, independent of sex hormones. Modulation of NO expression with MB does not improve survival or myocardial histological injury in either sex. PMID- 21636202 TI - Strong and weak aspects of an established post-resuscitation treatment protocol-A five-year observational study. AB - AIM OF STUDY: Favourable hospital survival increased from 26% to 56% in the implementation phase of a new standard operating procedure (SOP) for treatment after out-of hospital cardiac arrest (OHCA) in 2003. We now evaluate protocol adherence and survival rates after five years with this established SOP. METHODS: This observational study is based on prospectively collected registry data from all OHCA patients with cardiac aetiology admitted with spontaneous circulation to Ulleval Hospital between September 2003 and January 2009. Three patient categories are described based on early assessment in the emergency department: conscious, comatose, and comatose patients receiving only palliative care, with main focus on comatose patients receiving active treatment. RESULTS: Of 248 patients, 22% were consciousness on admission, 70% were comatose and received active treatment, while 8% received only palliative care. Favourable survival from admittance to discharge remained at 56% throughout the study period. Among actively treated patients 83% received emergency coronary angiography and 48% underwent subsequent percutaneous coronary intervention. In this cohort 63% had an acute myocardial infarction, ten of whom did not receive emergency coronary angiography. Among actively treated comatose patients, 6% survived with unfavourable neurology, while 51% of the deaths followed treatment withdrawal after prognostication of severe brain injury. CONCLUSION: The previously reported doubling in survival rate remained throughout a five-year study period. Establishing reliable indication for emergency coronary angiography and interventions and validating prognostication rules in the hypothermia era are important challenges for future studies. PMID- 21636203 TI - Pre-hospital cooling for out-of-hospital cardiac arrest--more research required. PMID- 21636205 TI - Maternal caffeine consumption and sine causa recurrent miscarriage. AB - OBJECTIVE: The objective of this study was to examine whether the risk of sine causa recurrent miscarriage is associated with caffeine consumption during the periconceptional period and early gestation after controlling for pregnancy related symptoms. STUDY DESIGN: A retrospective case-control study was conducted in the Department of Obstetrics and Gynecology, University of Turin between 2008 and 2009. Fifty-two sine causa recurrent miscarriers and 260 healthy pregnant women were assessed. Data were analyzed using SPSS 17 for Windows. RESULTS: Caffeine consumption during the periconceptional period and early gestation was higher in sine causa recurrent miscarriers compared to healthy pregnant women. Moreover, each caffeine intake of 100mg/day was associated with an increased odds ratio for sine causa recurrent miscarriage of 2724 (p for trend 0.001; 95% confidence interval [CI], 2.715-2.733), after adjusting for relevant confounding covariates. CONCLUSION: Caffeine intake may increase the risk of sine causa recurrent miscarriage regardless of pregnancy-related symptoms and relevant covariates (such as age and tobacco use). PMID- 21636204 TI - Haplotype-based association of ACE I/D, AT1R 1166A>C, and AGT M235T polymorphisms in renin-angiotensin-aldosterone system genes in Korean women with idiopathic recurrent spontaneous abortions. AB - OBJECTIVES: The objective of this study was to determine whether or not the angiotensin-converting enzyme insertion/deletion (ACE I/D), angiotensin II type 1 receptor (AT1R), and angiotensinogen (AGT) gene polymorphisms are associated with idiopathic recurrent spontaneous abortions (RSAs) in Korean women. STUDY DESIGN: A total of 251 patients with unexplained consecutive pregnancy losses, and 126 healthy controls with at least one live birth and no history of pregnancy loss. RESULT: The odds ratios (ORs) of the ACE ID (OR=2.423; 95% confidence interval (CI)=1.417-4.142; p=0.001) and the ACE II (OR=2.050; 95% CI=1.143-3.675; p=0.018) for the ACE DD genotype were significantly different between patients with idiopathic RSA and controls; however, there were no significant differences between patients and controls with respect to the AT1R 1166A>C and AGT M235T polymorphisms. In a haplotype-based analysis of I-A (p=0.010), D-A (p=0.004), I-A T (p=0.033), D-A-T (p=0.0005), and D-C-T (p=0.013) polymorphism pairs with synergistic effects derived by the MDR method in patients and in controls showed significant results. CONCLUSION: This study suggests that ACE, AT1R and AGT polymorphisms and haplotypes are a genetic determinant for the risk of idiopathic RSA in Korean women. PMID- 21636206 TI - Late-onset maternal mortality after amniocentesis. PMID- 21636207 TI - A successful case of abdominal radical trachelectomy for cervical cancer during pregnancy. PMID- 21636208 TI - Vaginal prolapse repair using the Prolift kit: a registry of 100 successive cases. AB - OBJECTIVES: To evaluate anatomical, functional outcomes and complications inherent to the treatment of pelvic organ prolapse by implantation of polypropylene mesh, using the Prolift kit. MATERIALS AND METHODS: Single-center observational study of 100 successive patients enrolled in a registry, who underwent Prolift prolapse repair. Data on prior treatments, associated procedures and per- and post-operative complications were collected and the patients were seen after 2, 6 and 12 months. Anatomical outcomes were assessed using the International Continence Society Pelvic Organ Prolapse Quantification (ICS POP-Q) system. Function was assessed in terms of urinary and digestive problems, and impact on sex life. Failure was defined as relapse of POP-Q Grade 2 or more. RESULTS: Of the 100 patients, 32 had an anterior, 14 a posterior, 54 a total Prolift; 53 had concomitant urinary incontinence surgery. At 6 and 12 months respectively, 8% and 12% of the patients were lost to follow-up. Mean operating time was 39.8 min. With respect to peri-operative complications, there was no bladder or rectal damage but three patients experienced bleeding (>= 300 ml) without needing transfusion. POP-Q Grades before surgery were: Grades III-IV cystocele in 65.7% of the anterior Prolift patients; Grades II-III rectocele in the posterior; and 77.8% of total were Grade III and 11.1% Grade IV. The incidence of recurrence was 3.6% at Month 6 and 10.2% at Month 12. Significant (p<0.05) improvements were seen in median scores for the various POP-Q items. With respect to functional problems, stress urinary incontinence was cured in 92% of the patients but 7.7% reported new-onset urinary incontinence after one year. One case of vaginal exposure after one year was observed and major or symptomatic mesh retraction was observed in 8%. New-onset dyspareunia was reported by 11.1% of the patients. CONCLUSIONS: These results confirm the feasibility of using the Prolift kit in the repair of prolapse via a vaginal approach and the low per- and post-operative morbidity associated with that technique. Nevertheless, longer term evaluation is required to confirm the results. PMID- 21636210 TI - Influence of the conserved active site residues of histidyl tRNA synthetase on the mechanism of aminoacylation reaction. AB - The relation between the conservation of active site residues and the molecular mechanism of aminoacylation reaction is an unexplored problem. In the present paper, the influences of the conserved active site residues on the reaction mechanism as well as the electrostatic potential near the reaction center are analyzed for Histidyl tRNA synthetase from Escherichia coli, Thermus thermophilus and Staphylococcus aureus. While the primary structures show both convergence as well as divergence, the secondary level structures of the active sites of the three species show considerable conservation in the respective structural organizations. The conserved active site residues near the reaction center, which have a major role in the reaction mechanism and catalysis, retain their specific position and orientation relative to the substrate in the three species. In order to understand the influence of different conserved and nonconserved residues near the reaction center, two different models are considered. First, a large model of active site with the substrates, Mg(2+) ions and water is constructed in which the first shell residues (including both conserved as well as nonconserved) near the reaction center are studied. From the large model, a smaller model is constructed for reaction path modeling individually for three species. Validation of the smaller model is carried out by comparing the energy surfaces of large and small models as a function of reaction coordinates. Further, the electrostatic potential near the reaction center for the large and small model are compared. The transition state structures of the activation step of aminoacylation reaction for E. coli, T. thermophilus and S. aureus are calculated using the combined ab initio/semi-empirical calculation. The similarity of the energy profiles as a function of the relevant reaction coordinate and the orientation of the catalytic residue, Arg259, indicate that the reaction mechanisms are identical which are guided by the strikingly similar structural pattern formed by conserved residues for three species. The energy surfaces have close resemblance in three species and present a clear perspective that how the reaction proceeds with the aid of different conserved residues. The study of electrostatic potential confirms this view. The present study provides an understanding of the relationship between the conservation of residues and the efficient reaction mechanism of aminoacylation reaction. PMID- 21636209 TI - E. coli SSB tetramer binds the first and second molecules of (dT)(35) with heat capacities of opposite sign. AB - We have previously shown that formation of a 1:1 fully wrapped complex of Escherichia coli SSB tetramer with (dT)(70) displays a temperature-dependent sign reversal of the binding heat capacity (DeltaC(P)). Here we examine SSB binding to shorter oligodeoxynucleotides ((dX)(35)) to probe whether this effect requires binding of one or two (dX)(35) molecules per SSB tetramer. We find that the DeltaC(P) for the first molecule of (dX)(35) is always negative. However, a sign reversal of DeltaC(P) from negative to positive occurs with increasing temperature for binding of the second (dX)(35). This striking behavior of DeltaC(P) for the second (dX)(35) appears linked to conformational changes within the ssDNA-SSB complex that are required to form a fully wrapped (SSB)(65) binding mode. These results also underscore that binding heat capacities of macromolecular interactions have multiple origins that cannot be understood simply on the basis of examining static structures. PMID- 21636211 TI - Greening the greenhouse grower. A behavioral analysis of a sector-initiated system to reduce the environmental load of greenhouses. AB - Growing plants and flowers in greenhouses is a commercial activity that imposes a burden on the environment. Recently a system of registration, control, and licensing has been developed by the sector of greenhouse growers in the Netherlands, acknowledged by the state. The current study was executed to understand the achievements of the greenhouse growers within this system. We applied a social-cognitive model to understand intentions to reduce emissions and predict actual pesticide use. The social-cognitive concepts from the model were measured in a questionnaire that was completed by 743 greenhouse growers. Factual information on these growers' pesticide use in 2004 and 2005 was provided by the agency responsible for the program, and could be linked to the questionnaire data. Using structural equation modeling we found that intention to reduce emission was primarily predicted by the attitude, descriptive norm, and self efficacy. Actual pesticide use was predicted by the interaction of intention and response efficacy. Results can be used to improve communication with growers, focusing on the influential determinants of intention and behavior. PMID- 21636212 TI - Kinetics of forced aerated biodegradation of digested sewage sludge-reed mixtures at different temperatures. AB - This work presents a kinetic analysis of the aerobic biodegradation of anaerobically digested sewage sludge and dried reed mixtures at different temperatures. Batch experiments were conducted in laboratory-scale reactors with temperature (T) control and forced aeration of the solid mixture. The biowaste mixture was treated at four different temperatures: 25, 40, 50 and 60 degrees C, with moisture controlled and samples taken weekly for carbon (C) and volatile solids (VS) measurements. The duration of experiments was either 90 d (at 25 degrees C) or 60 d (at 40, 50 and 60 degrees C). Two different kinetic models were used to fit the carbon mineralisation curves: the 2C model, which considers two organic fractions (biodegradable and non-biodegradable) and the 3C model, which considers three fractions (easily biodegradable, slowly biodegradable and non-biodegradable). In both cases, the kinetic rate constants were calculated by mathematical fitting and were compared with previously reported values. The temperature influence on the rate constants was studied for both models using a T dependent equation. The calculated kinetic rate constants were in agreement with previously published values, and good fitting of the experimental data was obtained with both models. Similar rate constant values were obtained for mineralisation of the biodegradable fraction (2C model) and the easily biodegradable fraction (3C model). The rate constants for the slowly biodegradable fraction (3C model) were much lower. A good correlation between rate constants and T was observed. Different optimum temperature values were obtained for each rate constant depending on which carbon fraction was degraded. The T-dependent rate constant values obtained could be used for modelling the C mineralisation of real variable-temperature composting processes. PMID- 21636213 TI - The estrogen receptor beta agonist diarylpropionitrile (DPN) inhibits medulloblastoma development via anti-proliferative and pro-apototic pathways. AB - Gender-related differences in medulloblastoma (MB) development have been reported with a higher incidence in males (slightly above 60%) than in females, female gender being also a significantly favorable prognostic factor in MB. The present study focused on the evaluation of the mechanisms by which estrogens protect against MB formation. To this end, we used a well characterized mouse model of MB - the Patched1 heterozygous mice. Ovariectomized mice were treated with 2,3-bis(4 hydroxyphenyl)-propionitrile (DPN), a highly potent ERbeta agonist, or 4,4',4"-(4 propyl-[1H]-pyrazole-1,3,5-triyl) trisphenol (PPT), a highly potent ERalpha agonist. Our results show that the ERbeta selective agonist DPN significantly inhibits development of MB preneoplastic lesions when compared with untreated ovariectomized mice, restoring the final incidence to that observed in the intact controls, and that these effects were achieved via activation of anti proliferative and pro-apototic pathways. On the other hand, the ERalpha selective agonist PPT did not influence MB tumorigenesis relative to untreated ovariectomized mice. PMID- 21636214 TI - Stress-induced hyperalgesia is associated with a reduced and delayed GABA inhibitory control that enhances post-synaptic NMDA receptor activation in the spinal cord. AB - GABA and glutamate are both affected by stress and are involved in nociception. Thus, we determined whether stress-induced enhancement of inflammatory hyperalgesia is mediated by an imbalance between glutamate and GABA neurotransmission. Male rats were subjected daily to 10 to 20 minutes per day of either forced swimming (FS) or sham swimming for 3 consecutive days; nonconditioned rats served as controls. Some rats were treated i.p. with ketamine (5 mg/kg), diazepam (2 mg/kg), flumazenil (0.1 mg/kg), or vehicle (0.9% NaCl), 30 to 60 minutes before each conditioning session or nociception assessment. Pain behavior, spinal nociceptive neuronal activation and GABA and glutamate release were respectively evaluated by the formalin test, the expression of c-Fos and in vivo microdialysis of superficial laminae of the lumbar spinal cord, 48 hours after the last conditioning session. Nitric oxide metabolites (NO(x)) were determined as markers of post-synaptic NMDA receptor activation. FS stress enhanced formalin-induced hyperalgesia, increased pain-elicited c-Fos expression, decreased basal and delayed pain-induced GABA release, and increased basal and induced glutamate release. Hyperalgesia and c-Fos overexpression were blocked only by prestress treatment with diazepam and post-stress treatment with ketamine, whereas changes in GABA and glutamate release were reversed by prestress treatment with diazepam. Diazepam effects were blocked by flumazenil. NO(x) increased in lumbar spinal cord of FS rats by a mechanism antagonized by ketamine. Thus, stress-induced hyperalgesia is initiated by a decreased and delayed GABA release and GABA-A receptor activation, whereas it is maintained by increased glutamate release and NMDA glutamate receptor activation at the spinal level. PMID- 21636215 TI - The use of larvae therapy to debride full thickness burns in the anaesthetically unfit patient: the Nottingham experience. PMID- 21636216 TI - Adolescents' self-reported reasons for using nicotine replacement therapy products: a population-based study. AB - BACKGROUND: Available research provides evidence that adolescents use nicotine replacement therapy (NRT) products. Yet, little is known about reasons and motives behind their use. The present study examined the reasons for NRT use among 14-18-year-old Finnish adolescents. METHOD: A national Adolescent Health and Lifestyle Survey was conducted in Finland in 2009 (N=4834, response rate 55%). Main measures were prevalence of NRT use, self-reported reasons for using NRT and smoking status. RESULTS: Overall, 10% had used NRT. Boys used NRT more often than girls (11.5% versus 8.7%, p<.001). The three most commonly reported reasons were 'just try' (56%), 'to quit' (33%) and 'smoking not possible' (24%). "Just try" was the most common reason given by non-smokers/experimental smokers whereas daily/occasional smokers used NRT mainly for quitting purposes and when smoking was impossible. CONCLUSIONS: These findings suggest that when planning treatment plans for adolescent smokers, health care personnel should pay particular attention to adolescents' primary reasons and motives for using NRT before suggesting its use. PMID- 21636217 TI - Concerns about dextromethorphan as a potential rapid-acting antidepressant. PMID- 21636218 TI - Is Gilbert syndrome a new risk factor for breast cancer? AB - Patients with Gilbert syndrome have an impaired function of the enzyme UGT1A1, responsible for the degradation of 4-OH-estrogens. These elements are produced by the degradation of estrogens and are well-known carcinogens. In theory, patients with Gilbert syndrome accumulate 4-OH-estrogens and, therefore, might have a higher risk for breast cancer, especially when exposed to higher levels of estrogens. If this theory is true, a new risk group for breast cancer would be described, producing new insights in breast carcinogenesis. PMID- 21636219 TI - On-line determination and control of fat content in batches of beef trimmings by NIR imaging spectroscopy. AB - An NIR imaging scanner was calibrated for on-line determination of the fat content of beef trimmings. A good calibration model was obtained for fat in intact beef (R=0.98, RMSECV=3.0%). The developed model could be used on single pixels to get an image of the fat distribution, or on the average spectrum from each trimming/portion of trimmings passing under the scanner. The fat model gave a rather high prediction error (RMSEP=8.7%) and a correlation of 0.84 when applied to 45 single trimmings with average fat content ranging from 1.6 to 49.3% fat. Test measurements on streams of trimmings making up batches varying from 10 to 24 kg gave a much lower prediction error (RMSEP=1.33%). Simulations based on true measurements indicate that the RMSEP decreases with increasing batch size and, for the present case, reached about 0.6% for 100 kg batches. The NIR scanner was tested on six batches of intact trimmings varying from 145 to 210 kg and gave similar fat estimates as an established microwave system obtained on the ground batches. The proven concept should be applicable to on-line estimation of fat in trimmings in order to determine the batch fat content and also to control the production of batches to different target fat levels. A possible requirement for the concept to work properly is that the trimming or layer of trimmings on the belt is not too thick. In this study maximum thickness was about 8 cm. Thicker trimmings might be measured, but careful hardware adjustments are then required. PMID- 21636220 TI - Effects of kimchi ethanolic extracts on oxidative stability of refrigerated cooked pork. AB - The antioxidant effect of different kimchi extracts in cooked ground pork during storage for 14 days at 4 degrees C was studied. Cooked ground pork was treated with ascorbic acid, BHT, baechu kimchi (BK), got kimchi (GK), puchu kimchi (PK), and white kimchi (WK) and compared to cooked ground pork without antioxidant. Radical scavenging and chelating activities of kimchi extracts were in the order: GK>PK>BK>WK. Total phenolic contents and flavonoid contents ranged from 32.52 to 46.73 mg of GAE/g and 5.87 to 25.58 mg quercetin/g, respectively. Significantly (P<0.05) lower values of TBARS, peroxide values, and hexanal contents were obtained for GK treated samples compared with cooked pork without antioxidant during refrigerated storage. GK showed good antioxidant activity and was significantly different (P<0.05) from the other treatments. Based on these findings, the natural antioxidants examined may have applications in the development of nutritionally enhanced meat products with enhanced shelf life. PMID- 21636221 TI - Effects of myosin heavy chain isoforms on meat quality, fatty acid composition, and sensory evaluation in Berkshire pigs. AB - The main objective of this study was to investigate the effects of myosin heavy chain (MHC) isoforms on meat and sensory quality in Berkshire pigs. A total of 85 pigs were evaluated, and muscle samples were taken for the analyses of MHC isoform, meat quality, fatty acid composition, and sensory evaluation. Content of the MHC slow isoform was significantly correlated with pH(24h) (r=0.26, P<0.05) and drip loss (r=-0.32, P<0.01), although the content of MHC isoforms showed limited relationships with individual fatty acids. In the case of sensory evaluation of meat by a trained panel test, the MHC fast/slow ratio was correlated with the juiciness (r=-0.33, P<0.01), off-flavor (r=0.34, P<0.01), tenderness attributes (r=-0.43 to -0.47). These results imply that the content of MHC isoforms can influence various aspects of quality including pork and sensory quality in Berkshire pigs. PMID- 21636222 TI - A novel solution for venous congestion following digital replantation: a proximally based cross-finger flap. AB - Digital replantations are often complicated by problems of venous congestion. Conservative management is not always successful. Furthermore, the skin edge around the replanted digit is frequently inflamed and necrotic, leading to difficulties in restoring venous flow by direct venous anastomosis or interpositional vein grafts. We introduce a novel solution using the proximally based cross-finger flap. We used this flap in 10 patients who had venous congestion with inflamed, necrotic skin at their digital replant site. Their initial injuries were amputation injuries. The flap had an average length of 3.98 cm and width of 2.59 cm and was harvested from the dorsum of the adjacent, uninjured digit. There was only 1 failure, due to massive crush injury. Of the remaining 9 cases, 7 met or exceeded the sensory threshold (Semmes-Weinstein monofilament test). The 2-point discrimination test was less than 6 mm in 8 cases. Three patients complained of residual pain (based on the Michigan Hand Outcomes Questionnaire), and only 1 was unsatisfied with the appearance. The proximally based cross-finger flap is pedicled and requires only a single level of venous anastomosis distally, leading to a higher success rate. It offers a simple yet effective solution for venous congestion. PMID- 21636223 TI - Open fractures of the distal radius: the effects of delayed debridement and immediate internal fixation on infection rates and the need for secondary procedures. AB - PURPOSE: There are few clinical data evaluating the outcome of surgery for open distal radius fractures based on treatment method. Specifically, the major contributing factors to infection are largely unknown. The purpose of this study is to determine the effect of early versus delayed debridement and the choice of initial external versus internal fixation on infection rates and the need for secondary procedures. METHODS: Thirty-two patients with open distal radius fractures were identified from a database. Ten debridements were early (<6 h after hospital admission), and 22 debridements were delayed (>6 h after hospital admission). There were 10 treating surgeons for the 32 patients in this study. Based on the attending surgeon's preference and experience, 20 fractures were treated with external fixation, 7 with plating, and 5 with planned staged conversion from external fixation to plating. The cohort included 19 grade I, 11 grade II, and 3 grade IIIA open injuries. RESULTS: There were no infections, regardless of the time to debridement or the use of immediate plating. Other complications requiring secondary procedures occurred more frequently in patients treated with a planned staged conversion from external fixation to plating than in the patients treated with either external fixation or plating. CONCLUSIONS: We did not encounter infections for grade I and grade II open distal radius fractures, and infections do not appear to be related to either the time to debridement or the initial type of fracture fixation. Plating might be safe at the initial debridement, but temporary external fixation with a staged conversion to plating increases the risk of complications, which necessitates corrective secondary procedures. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic III. PMID- 21636224 TI - Capsular imbrication for posttraumatic instability of the distal radioulnar joint. AB - PURPOSE: To analyze functional and subjective outcomes of patients with posttraumatic dorsal instability of the distal radioulnar joint (DRUJ) treated by a dorsal capsular imbrication. METHODS: We reviewed 11 patients (7 men and 4 women) with posttraumatic instability of the DRUJ who were treated by a capsular imbrication. The patients ranged in age from 18 to 48 years (average, 33 y). The duration of symptoms before surgery ranged from 3 weeks to 6 months (average, 5 mo). We evaluated subjective outcome measurements such as the Disabilities of the Arm, Shoulder, and Hand (DASH) questionnaire, questions regarding symptoms of pain and instability, and overall satisfaction. Objective outcome measurements were physical examination, standard radiographs, and a review of any surgical complications. Patients were asked whether they experienced wrist tenderness with applied pressure, swelling, scar sensitivity, or pain. We determined functional outcome using the Modified Mayo Wrist Score including range of motion and grip strength. The score was used to evaluate the surgical technique and enable comparison with other surgical methods used to treat instability of the DRUJ. RESULTS: After surgery, the total mean DASH score was 15 points. Range of motion, grip strength, pain, and overall function resulted in a mean Modified Mayo Wrist Score of 88 points, and therefore a good functional outcome. Three patients were noted to have persistent mild DRUJ instability on the stress test but did not have symptoms. The only intraoperative or postoperative complications observed were transient paresthesias on the ulnar side of the hand. CONCLUSIONS: Capsular imbrication for posttraumatic instability of the DRUJ is a highly effective procedure with few complications and good functional and subjective results even in cases with concomitant healed fractures of the radius or ulnar styloid. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic IV. PMID- 21636225 TI - Changes in illicit drug consumption patterns in 2009 detected by wastewater analysis. AB - BACKGROUND: Wastewater analysis can provide estimates of illicit drug (ID) consumption in local communities. METHODS: We used repeated raw wastewater analysis in urban wastewater treatment plants to estimate loads of cocaine, heroin, methamphetamine, and cannabis consumed daily by the inhabitants of two cities in Northern Italy, Milan and Como, from 2005 to 2009. RESULTS: Daily cocaine loads did not change in Milan from 2005 to 2008 but fell 45% in 2009 (ANOVA, p<0.001, followed by Tukey-Kramer HSD test [2009 vs. others], p<0.05), and there was a similar drop in Como (41%, p<0.0001, t-test). Heroin also fell from 2008 to 2009 in Milan (66%, ANOVA, p<0.001, followed by Tukey-Kramer [2009 vs. others], p<0.05) and Como (26%, p=0.017, t-test). However, methamphetamine, which had risen in Milan from 2005 to 2008, rose further in 2009 (Kruskal-Wallis test, p<0.001, followed by Steel-Dwass [2009 or 2008 vs. previous], p<0.05), and cannabis, which was falling from 2005 to March 2009, rose again in September 2009 (40%, p=0.027, t-test). CONCLUSIONS: Results suggest a trend toward a decrease in consumption of costly ID, such as cocaine and heroin. This might be due to a reduction in the number of consumers and/or to a change in their behaviour since there was also an increase in the consumption of less expensive ID. This itself might reflect a drop in consumers' money supply, caused by the economic crisis. Wastewater analysis was useful to estimate ID consumption levels in local communities in real time and promptly identify changes in trends. PMID- 21636226 TI - Strategies for characterizing complex phenotypes and environments: general and specific family environmental predictors of young adult tobacco dependence, alcohol use disorder, and co-occurring problems. AB - BACKGROUND: Defining phenotypes in studies of tobacco and alcohol misuse is difficult because of the complexity of these behaviors and their strong association with each other and with other problem behaviors. The present paper suggests a strategy for addressing this issue by conceptualizing and partitioning variance in phenotypes into either general or substance/behavior-specific. The paper also applies the general or substance/behavior-specific conceptualization to environmental predictors of tobacco and alcohol misuse and other problem behaviors. METHODS: Data were drawn from the Seattle Social Development Project, a contemporary, ethnically diverse and gender-balanced longitudinal panel including 808 participants. Latent variable modeling was used to partition variance in young adult (age 24) nicotine dependence, alcohol abuse and dependence, illicit drug abuse and dependence, involvement in crime, and engagement in HIV sexual risk behavior into general problem behavior and behavior specific variance. Similarly, measures of general, drinking-specific, and smoking specific adolescent family environment were constructed. RESULTS: Consistent with expectations, more positive general family environment during adolescence was associated with lower levels of shared variance in problem behaviors at age 24, but not with unique variance in tobacco or alcohol use disorder. Higher levels of family smoking and drinking environments during adolescence, however, were positively associated with unique variance in tobacco and alcohol use disorder, respectively, but did not predict shared variance in problem behaviors. CONCLUSIONS: Results support the utility of the proposed approach. Ways in which this approach might contribute to future molecular genetic studies are discussed. PMID- 21636227 TI - Survival of faecal indicator bacteria in treated pig manure stored in clay covered heaps in Vietnam. AB - Small-scale pig farmers in Vietnam typically store manure in heaps covered by a layer of clay. However, the hygienic quality of manure applied to agricultural soil is uncertain as storage time is determined by the need for soil fertilizer. This study aimed to assess how the storage of pig manure added straw, lime stone or urea in heaps covered by clay affected the survival of faecal bacteria. E. coli was reduced from 4 log(10)CFU/g to below the detection limit within 2 weeks with 2% (w/w) urea (CO(NH(2))(2)) whereas other amendments were associated with limited E. coli reductions. Enterococcus spp. were not reduced in any heaps indicating possible re-growth and that enterococci should be used as hygiene indicators with caution. Follow-up research should document the practical conditions under which the farmer practice of storing manure with a clay cover can be utilized to obtain a bactericidal effect of ammonia and a safe product with high fertilizer value. PMID- 21636228 TI - Characteristics of cannabinoids composition of Cannabis plants grown in Northern Thailand and its forensic application. AB - The Thai government has recognized the possibility for legitimate cultivation of hemp. Further study of certain cannabinoid characteristics is necessary in establishing criteria for regulation of cannabis cultivation in Thailand. For this purpose, factors affecting characteristics of cannabinoids composition of Thai-grown cannabis were investigated. Plants were cultivated from seeds derived from the previous studies under the same conditions. 372 cannabis samples from landraces, three different trial fields and seized marijuana were collected. 100g of each sample was dried, ground and quantitatively analyzed for THC, CBD and CBN contents by GC-FID. The results showed that cannabis grown during March-June which had longer vegetative stages and longer photoperiod exposure, had higher cannabinoids contents than those grown in August. The male plants grown in trial fields had the range of THC contents from 0.722% to 0.848% d.w. and average THC/CBD ratio of 1.9. Cannabis in landraces at traditional harvest time of 75 days had a range of THC contents from 0.874% to 1.480% d.w. and an average THC/CBD ratio of 2.6. The THC contents and THC/CBD ratios of cannabis in second generation crops grown in the same growing season were found to be lower than those grown in the first generation, unless fairly high temperatures and a lesser amount of rainfall were present. The average THC content in seized fresh marijuana was 2.068% d.w. while THC/CBD ratios were between 12.6 and 84.09, which is 10-45 times greater than those of similar studied cannabis samples from the previous study. However, most Thai cannabis in landraces and in trial fields giving a low log(10) value of THC/CBD ratio at below 1 may be classified as intermediate type, whereas seized marijuana giving a higher log(10) value at above 1 could be classified as drug type. Therefore, the expanded information provided by the current study will assist in the development of criteria for regulation of hemp cultivation in Thailand. PMID- 21636229 TI - A cupulolith repositioning maneuver in the treatment of horizontal canal cupulolithiasis. AB - OBJECTIVE: We evaluated outcomes and their significance of a new treatment method for horizontal canal cupulolithiasis that could be applied regardless of the side of the cupula where otoliths are attached. METHODS: Consecutive 78 patients who showed persistent apogeotropic horizontal canal positional vertigo (horizontal canal cupulolithiasis) were enrolled, and they were treated with the new cupulolith repositioning maneuver. RESULTS: Horizontal semicircular canal cupulolithiasis was alleviated in 97.4% of patients, after an average of 2.1 repetitions of the maneuver. Otoliths were suspected to be attached to the canal side of the cupula in 30 cases and the utricular side in 44 cases. CONCLUSION: The cupulolith repositioning maneuver is an effective method for treating horizontal canal cupulolithiasis. It may also provide an insight into the side of the cupula where otoliths are attached. PMID- 21636230 TI - Efficacy of combined treatment with S-carboxymethylcysteine (carbocisteine) and clarithromycin in chronic rhinosinusitis patients without nasal polyp or with small nasal polyp. AB - OBJECTIVE: In Japan, fourteen-membered ring macrolides, antibacterial agents, and S-carboxymethylcysteine (SCMC; carbocisteine), a mucolytic, are commonly used to treat chronic rhinosinusitis (CRS), and they are also used in combination. However, no large-scale randomized study has examined the effects of these pharmacotherapies. The aim of this study is to evaluate the effect of combined administration of clarithromycin (CAM), a fourteen-membered ring macrolide, and SCMC, compared with CAM single therapy. METHODS: Patients with CRS were centrally registered and randomly assigned to treatment with CAM (200mg/day) alone (monotherapy group) or CAM (200mg/day) in combination with SCMC (1500mg/day; combination group) for 12 weeks. We assessed the clinical efficacy of the treatments using measures of subjective symptoms and objective findings, health related quality of life (HRQOL) determined by the 20-Item Sino-Nasal Outcome Test (SNOT-20) score and computed tomography (CT) score. RESULTS: Four hundred twenty five subjects were enrolled (combination group, 213; monotherapy group, 212). At week 12 of treatment, the rate of effectiveness was significantly higher in the combination group (64.2%) compared with the monotherapy group (45.6%; P=0.001). In addition, objective findings, including characteristics of nasal discharge (P=0.008) and post-nasal discharge (P=0.002) were significantly improved in the combination group. In both groups, SNOT-20 and CT scores were significantly improved from week 0 (P<0.001), and were not significantly different between groups. CONCLUSION: The results indicated that long-term combination therapy with SCMC at a dose of 1500mg/day and CAM at a dose of 200mg/day is effective for improving subjective symptoms and objective findings in adult patients with CRS. PMID- 21636231 TI - Adrenal neoplasms: Effectiveness and safety of CT-guided ablation of 23 tumors in 22 patients. AB - PURPOSE: To retrospectively evaluate the effectiveness and safety of image-guided ablation of adrenal tumors. MATERIALS AND METHODS: : This HIPAA-compliant study was IRB approved and informed consent was waived. From 5/1999-6/2008, 20 consecutive adrenal metastases (mean diameter 4.2cm; range, 2-8) and 3 hormonally active primary adrenal tumors (mean diameter 2.3cm; range, 1-4), including an aldosteronoma and 2 pheochromocytomas in 22 patients (14 men, 8 women; mean age 61 years; range 40-84) were ablated in 23 sessions. Bilateral metastases were treated in a single patient. Radiofrequency ablation was used to treat 16 adrenal metastases and the 3 hyperfunctioning tumors. Microwave ablation was used to treat 4 metastases. Successful treatment was defined as a lack of both enhancement on follow-up contrast enhanced CT and/or up-take on FDG PET-CT and for functioning tumors, resolution of biochemical abnormalities. RESULTS: Technical success was achieved in all sessions. Mean follow-up was 45.1 months (range, 1-91) Local tumor progression (focal enhancement at ablation site >=1cm in short axis) was detected in 4 of 23 tumors, two of which were identified bilaterally in a single patient prompting re-treatment. Of 19 patients with metastatic disease, 16 had fatal extra-adrenal disease progression, and 3 remain alive. Two of the 3 patients who underwent ablation of hyperfunctioning tumors remain alive, including the patient with an aldosteronoma who had recurrent symptoms 91 months post ablation. Intra-ablative hypertension occurred in 9% (2/23) of sessions and was successfully treated pharmacologically. CONCLUSION: Ablation of metastatic and hyperfunctioning adrenal tumors is safe and may provide local control and treatment of pathologic biochemical activity. PMID- 21636232 TI - T2WI MRI and MRI-MDCT correlations of the osteoporotic vertebral compressive fractures. AB - PURPOSE: The purpose of this study was to present the various vertebral bone signal and configuration patterns on T2WI MRI in osteoporotic benign fractures. The appearances were also assessed with thin slice MPR CT images. MATERIALS AND METHODS: 66 patients (43 women, 23 men; age range, 34-97 years; mean age, 76 years) with 100 vertebral compression fractures due to osteoporosis were included in our study. All cases are acute or subacute phase fractures which occurred within 1 month. Multiple fractures were observed in 29 patients, in 8 cases multiple concurrent fractures were proved by past radiograph or MRI. Three planes of T2WI (sagittal, coronal, axial) section images were correlated with the corresponding MDCT images. RESULTS: In our retrospective study, various signals and forms of vertebral body fractures were observed. The fracture zone (line) was classified 3 signal intensity patterns. (high; n = 24, low to intermediate; n = 16, low intensity; n = 38) on T2WI. The signal of the peri(para) fractured area were high (n = 6), intermediate to low (n = 41), low (n = 53)intensity. With CT correlative study, high, low to intermediate area were considered to be corresponded to the bone marrow edema, fibrous reparative tissue with/without the bony callus formation. The localization were as follows: sub-end plate band like lesion in 53, central in 16, anterior in 13, sub-endplate+anterior in 10, and complete (diffuse) in 8. In 10 cases, adjacent Schmorl's nodules were observed. CONCLUSION: The various vertebral body signal patterns were observed in osteoporotic compressive fracture on T2WI. By MRI-CT correlations, high signal and low signal area on T2WI corresponded to the bone marrow edema, and the fibrous tissue or callus formation respectively. PMID- 21636233 TI - Stability assessment of o/w parenteral nutrition emulsions in the presence of high glucose and calcium concentrations. AB - The purpose of the present study was to examine how the colloid stability features of o/w parenteral nutrition emulsions made with SMOFlipid (lipid emulsion based on soybean oil, medium chain triglycerides, olive oil and fish oil) will change in the presence of high concentration of calcium and glucose if usual micronutrients are also present, according to the needs of the clinical nutrition patient. Particle size analysis, zeta potential, dynamic surface tension measurements and light microscopic screening were carried out to evaluate the possible changes in the kinetic stability of the emulsions. Our results indicate that the higher glucose concentration of 15 or 20% could not compensate the emulsion-destabilizing effect of higher (5 mM) calcium concentration even in the presence of a modern fat emulsion. Therefore calcium demand of undernourished patient requiring 5 mM or higher final Ca2+ content in nutrient solution should be supplemented in another way. PMID- 21636234 TI - Identification of oxidative degradation impurities of olanzapine drug substance as well as drug product. AB - Impurities found in stressed and stability studies of olanzapine (polymorphic form-I) [1-7] in both drug substance and drug product are described. These impurities are identified as 4-(4-methyl-1-piperazinyl)-3-hydroxymethylidene-1H benzo[b][1,4]diazepine-2(3H)-thione (hydroxymethylidene thione) and (Z)-4-(4 methyl-1-piperazinyl)-3-acetoxymethylidene-1H-benzo[b][1,4]diazapine-2(3H)-thione (acetoxymethylidene thione). An oxidative degradation pathway of olanzapine, for the formation of these impurities, has been proposed. PMID- 21636235 TI - Content uniformity studies in tablets by NIR-CI. AB - Near infrared chemical imaging (NIR-CI) is attracting growing interest in pharmaceutical analysis by virtue of its ability to provide a wealth of information from a single sample. Among others, NIR-CI has enabled the determination of the quantitative composition and distribution of acetylsalicylic (ASA) from the analysis of commercial tablets. In this work, we analyzed ASA commercial tablets of four different brands purchased at local chemists. The nominal ASA concentration for the brands was calculated from the nominal content and averaged weight of tablets. The tablets were found to span an ASA concentration range of 71-82%, and to differ in size and composition between brands. The API content and its homogeneity distribution were determined by applying quantitative algorithm to global hyperspectral image of ten tablets. Multivariate curve resolution-alternating least squares (MCR-ALS) is used to quantify each pixel in the images to obtain appropriate concentration maps. No prior calibration or reference data were needed for quantitation and results are close to the nominal content used as reference. Application to an image for 10 tablets and an individual tablet quantitation of the API allowed us to obtain the Accepted Value (AV) as defined by the European Pharmacopoeia. We conclude that all brands meet the pharmacopoeia specifications. PMID- 21636236 TI - Shedding light on the dark side of doctor-patient interactions: verbal and nonverbal messages physicians communicate during error disclosures. AB - OBJECTIVE: Existing investigations on medical error disclosures have neglected the fact that a disproportionately large amount of the meaning in messages is derived from nonverbal cues. This study provides an empirical assessment of the verbal and nonverbal messages physicians communicate when disclosing medical errors to standardized patients. METHODS: Sixty hypothetical error disclosures by a volunteer sample of attending physicians were videotaped, coded, and statistically analyzed. RESULTS: Physicians used friendly, smooth, approaching and invested nonverbal styles as they disclosed medical errors to standardized patients. Female physicians smiled more and were more attentive to patients than male physicians, and physicians tended to exhibit more positive affect in the form of facial pleasantness toward angry female patients than toward angry male patients. Furthermore, physicians touched and smiled at patients more frequently at the beginning and at the end of their error disclosures, and displayed decreased attentiveness and interactional fluency. CONCLUSION: Future research needs to examine which disclosure styles patients perceive as competent, and to assess their causal impacts on objective and relational disclosure outcomes. PRACTICE IMPLICATIONS: This study provides an important baseline understanding of medical error disclosures that is essential for the successful implementation of empirically based training programs. PMID- 21636237 TI - Survey of family physicians' perspectives on management of immigrant patients: attitudes, barriers, strategies, and training needs. AB - OBJECTIVE: Immigrants in Canada form a significant portion of the population and have unique and complex health needs. This study was undertaken to evaluate family physicians' perspectives on the care of this population. METHODS: Questionnaires were distributed to family physicians in Montreal (n=598). The main outcomes of interest were attitudes of family physicians to care of immigrants including barriers perceived, resources and strategies used to accommodate immigrant patients, as well as physicians' training in immigrant care. RESULTS: Family physicians find communication difficulties to be the key barrier and would like to see the access to interpreters improved. Very few physicians make use of professional interpreters. Only a minority of physicians have received specific cross-cultural competence training but those who have seem to provide better quality of care. CONCLUSIONS: Knowledge of physician perspectives is an essential element on which to base interventions to improve the quality of care to this population. PRACTICE IMPLICATIONS: Physicians should be reminded of the importance of using professional interpretation services in multi-lingual encounters. Cross-cultural training should be further advanced in Canadian medical curricula. PMID- 21636238 TI - Evaluating preventable adverse safety events after elective lower extremity procedures. AB - BACKGROUND: Improving patient safety has become a national priority. Patient safety indicators (PSIs) are validated tools to identify potentially preventable adverse events. No studies currently exist for evaluating lower extremity (LE) vascular procedures and the occurrence of PSIs. METHODS: The Nationwide Inpatient Sample (2003-2007) was queried for elective LE angioplasty (endo) and bypass (open). PSIs appropriate to surgery were analyzed by chi(2) and logistic regression analyses. RESULTS: A total of 226,501 LE interventions (104,491 endo; 122,010 open) were evaluated. The rate of developing any PSI was 7.74% (open) and 8.51% (endo), P < .0001. In the latter group, PSI9 (postoperative hemorrhage or hematoma) with the rate 4.74% was a predominant indicator that was associated with an almost three times greater likelihood of death in this group. PSI predictors included advanced age (odds ratio [OR], 1.64; 95% confidence interval [CI], 1.55-1.75 for oldest vs youngest patients), females (OR, 1.18; 95% CI, 1.13 1.22), blacks (OR, 1.10; 95% CI, 1.04-1.17), congestive heart failure (OR, 1.83; 95% CI, 1.72-1.96), diabetes mellitus (OR, 1.20; 95% CI, 1.12-1.28), renal failure (OR, 2.31; 95% CI, 2.14-2.50), hospital teaching status (OR, 1.21; 95% CI, 1.17-1.26), and larger hospitals (OR, 1.11; 95% CI, 1.05-1.17). PSI occurrence was associated with increased cost ($28,387 vs $13,278; P < .0001). CONCLUSIONS: Endovascular procedures were found to have lower mortality rates overall, but were found to have a greater number of safety events occur primarily due to bleeding complications in women and the elderly. PSIs were associated with advanced age, black race, and comorbidities. Adverse events added significant cost, occurred more frequently in teaching and large hospitals, and future organizational analysis may improve safety and reduce cost. PMID- 21636239 TI - Results of the Protege EverFlex 200-mm-long nitinol stent (ev3) in TASC C and D femoropopliteal lesions. AB - OBJECTIVES: This study investigated the results with primary stenting using the Protege EverFlex 200-mm-long self-expanding nitinol stent (ev3 Endovascular Inc, Plymouth, Minn) in femoropopliteal TransAtlantic Inter-Society Consensus (TASC) C and D lesions of at least 150 mm in length. METHODS: Between March 2008 and June 2009, 100 patients (66 men) presenting with 100 symptomatic TASC C and D femoropopliteal lesions were treated with at least one 200-mm-long Protege EverFlex stent. The intention of this study was to treat all lesions with as few stents as possible. The primary study end point was primary patency at 12 months, defined as the absence of hemodynamically significant stenosis on duplex ultrasound imaging (systolic velocity ratio <2.4) at the target lesion and without target lesion revascularization (TLR) <=12 months. Stent fracture occurrence was assessed at the 12-month follow-up by conventional x-ray imaging. RESULTS: Average patient age was 70 years. Preoperative symptom assessment reported 71 patients (71%) had claudication vs 29 (29%) with critical limb ischemia. Average lesion length was 242 mm (range, 160-450 mm), and 27 patients (27%) presented with popliteal involvement. A total of 158 Protege EverFlex stents were used to treat 100 lesions. Kaplan-Meier estimation reported a 12 month freedom from target lesion revascularization of 68.2% and a primary patency rate of 64.8%. Stent fractures occurred in six patients (6.0%) when x-ray images taken immediately after the procedure were compared with those taken after 1 year. CONCLUSIONS: The results of our Durability-200 study show an acceptable primary patency rate after 1 year was obtained in this patient cohort with TASC C and D femoropopliteal lesions. PMID- 21636240 TI - Late outcomes of balloon angioplasty and angioplasty with selective stenting for superficial femoral-popliteal disease are equivalent. AB - OBJECTIVE: Several trials have reported early superior patency of stenting over isolated angioplasty (plain old balloon angioplasty [POBA]) for infra-inguinal occlusive disease, yet long-term data are sparse. The purpose of this study was to contrast long-term clinical outcomes and costs of angioplasty alone vs angioplasty with selective stenting in the treatment of femoropopliteal occlusive disease. METHODS: Patients undergoing primary endovascular treatments of the native femoropopliteal arteries from 2002 to 2009 were divided into two groups, POBA alone or stenting based on final treatment received at their index procedure. Study end points included actuarial 5-year primary patency (using strict criteria of any hemodynamic deterioration or return of symptoms), 5-year limb salvage, and 5-year survival and hospital costs. RESULTS: Eight hundred twenty-four primary procedures were performed during the study interval; 517 (63%) were POBA and 307 (37%) were stenting. The mean follow-up duration was 33 months (range, 0-98 months). The indication for intervention in the stenting group was claudication in 71% of the patients, whereas the remaining 29% had critical limb ischemia (CLI). In the POBA cohort, the indication for treatment was claudication in 59% of the patients and CLI in the remaining 41%. A higher percentage of POBA lesions were TransAtlantic Inter-Society Consensus (TASC) II A & B when compared to stenting (91% POBA vs 73% stenting; P < .001). There was no difference in overall 5-year primary patency (POBA 36% +/- 3%; stenting 41% +/- 4%; P = .31), nor was there a difference in patients with claudication (POBA 42% +/- 4%; stenting 45% +/- 4%; P = .8). In patients with CLI, the 4-year primary patency was 27% +/- 5% (POBA) vs 36% +/- 8% (stenting), P = .22; the 4-year limb salvage was 80% +/- 4% (POBA) vs 90% +/- 5% (stenting), P = .18. There was no difference in survival between the two groups (claudication: 83% +/- 3% POBA vs 84% +/- 4% stenting at 5 years (P = .65), CLI: 44% +/- 4% POBA vs 49% +/- 6% stenting at 4 years (P = .40). Subgroup analysis by lesion anatomy showed similar primary patency between POBA and stenting for TASC II A & B lesions, while the primary patency was significantly higher at 5 years after stenting of TASC II C & D lesions (34% +/- 6% vs 12% +/- 9%; P < .05). Stenting increased the procedural cost by 57% when compared to POBA (P < .001) regardless of treatment indication. In addition, stenting added 45% (P < .001) to the overall hospital cost of patients treated for claudication. CONCLUSION: Stenting resulted in equivalent long-term outcomes compared to POBA when stratified by indications. However, stenting yielded statistically better primary patency in patients with TASC II C & D lesions. The lack of improved clinical outcomes and significantly higher cost of stenting supports a posture of selective use of stents (especially in TASC II A & B) in the endovascular treatment of femoropopliteal occlusive disease. PMID- 21636241 TI - The importance of expert feedback during endovascular simulator training. AB - OBJECTIVES: Complex endovascular skills are difficult to obtain in the clinical environment. Virtual reality (VR) simulator training is a valuable addition to current training curricula, but is there a benefit in the absence of expert trainers? METHODS: Eighteen endovascular novices performed a renal artery angioplasty/stenting (RAS) on the Vascular Interventional Surgical Trainer simulator. They were randomized into three groups: Group A (n = 6, control), no performance feedback; Group B (n = 6, nonexpert feedback), feedback after every procedure from a nonexpert facilitator; and Group C (n = 6, expert feedback), feedback after every procedure from a consultant vascular surgeon. Each trainee completed RAS six times. Simulator-measured performance metrics included procedural and fluoroscopy time, contrast volume, accuracy of balloon placement, and handling errors. Clinical errors were also measured by blinded video assessment. Data were analyzed using SPSS version 15. RESULTS: A clear learning curve was observed across the six trials. There were no significant differences between the three groups for the general performance metrics, but Group C made fewer errors than Groups A (P = .009) or B (P = .004). Video-based error assessment showed that Groups B and C performed better than Group A (P = .002 and P = .000, respectively). CONCLUSION: VR simulator training for novices can significantly improve general performance in the absence of expert trainers. Procedure-specific qualitative metrics are improved with expert feedback, but nonexpert facilitators can also enhance the quality of training and may represent a valuable alternative to expert clinical faculty. PMID- 21636242 TI - Modern PET imaging for paragangliomas: relation to genetic mutations. AB - AIM AND METHODS: Paragangliomas (PGL) are neural crest-derived tumours that are found along the autonomic neural network throughout the body and can be multiple and/or metastatic. Nuclear medicine imaging in combination with conventional imaging is required to fully delineate the extent of the disease. The performance of molecular imaging modalities is widely dependent on tumour biology. RESULTS: In the present paper we discuss the recent publications focused on the role of positron emission tomography (PET) imaging and the relationship between tracer uptake patterns and genetic mutations associated with the disease. CONCLUSION: Recent advances in genetic and molecular pathogenesis of PGL have allowed for the identification of new molecular diagnostic and therapeutic radiopharmaceuticals tailored to genetic abnormalities. However, the optimal diagnostic imaging algorithm remains to be determined. PMID- 21636243 TI - Sentinel lymph node biopsy to direct treatment in gastric cancer. A systematic review of the literature. AB - Gastric cancer is one of the main causes of cancer-related deaths around the world. The prevalence of early gastric cancer (EGC) among all gastric cancers of 45-51% in Japan, but only 7-28% in Western countries. The prevalence of EGC is growing partly because of better diagnostics and screening programmes. Possible treatment options for EGC treatment are expanded by the introduction of endoscopic mucosal resection and endoscopic submucosal dissection Therefore, detailed knowledge about nodal metastatic risk is warranted. We performed a systematic review of the literature concerning studies investigating the role of sentinel lymph node biopsy in EGCr and whether there is enough proof to introduce SLN as a part of treatment for EGC in the Netherlands. Several detection substances (dye or radiocolloid) and injection methods (submucosal or subserosal) are investigated. An overall sensitivity percentage of 85.4% was found. In comparison, high and clinically sufficient percentages were observed for specificity (98.2%), negative predictive value (90.7%) and accuracy (94%). Subgroup analyses showed that the combination of dye and radiocolloid detection substances is the best method for sentinel lymph node detection in early gastric cancer. However, the precise method of sentinel lymph node biopsy in EGC has to be determined further. Large, randomized series should be initiated in Europe to address this issue. PMID- 21636244 TI - Long-term follow-up reveals that ulceration and sentinel lymph node status are the strongest predictors for survival in patients with primary cutaneous melanoma. AB - PURPOSE: To assess the long-term outcome after sentinel lymph node biopsy (SLNB) in melanoma patients. METHODS: Between 1995-2009 450 melanoma patients underwent SLNB in a single center. Survival and prognostic factors were analyzed for 429 patients. RESULTS: Median age was 53 (range 11-84) years. Median Breslow thickness was 2.4 (range 1-20) mm and 36% were ulcerated melanomas. Median follow up time was 64.8 (range 2-174) months. A tumor-positive SLN was present in 140 patients (31%). Completion lymph node dissection (CLND) was performed in 119 patients and these patients were analyzed for recurrence and survival. 124 Patients (29%) relapsed during follow-up; 55 in the node-positive group who underwent CLND (55/119; 46%) and 69 in the node-negative group (69/310; 22%; p < 0.001). In the node-negative group 17 patients developed recurrence in the regional node field; false-negative rate 11%. On multivariate analysis strongest prognostic factors for disease free survival (DFS) were primary melanoma ulceration and SLN positivity (Hazard Ratio (HR) of 2.2 and 2.3; p < 0.001). For disease specific survival (DSS) the same was found to be true with an HR of 2.1 for ulceration and 2.0 for SLN positivity (p = 0.001 and p = 0.002 respectively). 10-Year DFS was 71% for node-negative patients compared with 48% for node positive patients (p < 0.001). 10-Year DSS was 77% for node-negative patients compared to 60% for node-positive patients (p < 0.001). CONCLUSIONS: This study shows a remarkably high percentage of tumor-positive SLN. The long-term follow-up data confirm that tumor-positive SLN patients have a worse DFS and DSS than tumor negative SLN patients. Ulceration and SLN status proved to be the strongest prognostic factors for long-term DFS and DSS. PMID- 21636245 TI - Anger suppression after imagined rejection among individuals with social anxiety. AB - Individuals experiencing high levels of social anxiety report frequent and intense anger. Yet, little is known about how they manage this emotion. Despite general tendencies towards anger suppression, subsets of individuals with social anxiety regulate anger through outward expression. In this study, we investigated rejection as an antecedent to anger, examined how and when individuals with high social anxiety suppress anger, and evaluated experiential avoidance (EA) as a moderator of the relationship between social anxiety and anger suppression. 170 undergraduate students described their responses to everyday social situations that were designed to elicit anger; several situations reflected instances of social rejection. Our results suggest that rejection was a potent source of anger for most people and that social anxiety predicted both anger and EA in response to imagined rejection. In addition, as evidence of a moderation model, individuals with low social anxiety and low EA reported the least anger suppression; no significant differences were found for individuals with high social anxiety. We discuss the implications for understanding the interface of social anxiety and anger. PMID- 21636246 TI - Observational studies in systematic [corrected] reviews of comparative effectiveness: AHRQ and the Effective Health Care Program. AB - OBJECTIVE: Systematic reviewers disagree about the ability of observational studies to answer questions about the benefits or intended effects of pharmacotherapeutic, device, or procedural interventions. This study provides a framework for decision making on the inclusion of observational studies to assess benefits and intended effects in comparative effectiveness reviews (CERs). STUDY DESIGN AND SETTING: The conceptual model and recommendations were developed using a consensus process by members of the methods workgroup of the Effective Health Care Program of the Agency for Healthcare Research and Quality. RESULTS: In considering whether to use observational studies in CERs for addressing beneficial effects, reviewers should answer two questions: (1) Are there gaps in the evidence from randomized controlled trials (RCTs)? (2) Will observational studies provide valid and useful information? The latter question involves the following: (a) refocusing the study questions on gaps in the evidence from RCTs, (b) assessing the risk of bias of the body of evidence of observational studies, and (c) assessing whether available observational studies address the gap review questions. CONCLUSIONS: Because it is unusual to find sufficient evidence from RCTs to answer all key questions concerning benefit or the balance of benefits and harms, comparative effectiveness reviewers should routinely assess the appropriateness of inclusion of observational studies for questions of benefit. Furthermore, reviewers should explicitly state the rationale for inclusion or exclusion of observational studies when conducting CERs. PMID- 21636247 TI - Discovery of low-affinity preproinsulin epitopes and detection of autoreactive CD8 T-cells using combinatorial MHC multimers. AB - Autoreactive cytotoxic CD8 T-cells (CTLs) play a key pathogenic role in the destruction of insulin-producing beta-cells resulting in type 1 diabetes. However, knowledge regarding their targets is limited, restricting the ability to monitor the course of the disease and immune interventions. In a multi-step discovery process to identify novel CTL epitopes in human preproinsulin (PPI), PPI was digested with purified human proteasomes, and resulting COOH-fragments aligned with algorithm-predicted HLA-binding peptides to yield nine potential HLA A1, -A2, -A3 or -B7-restricted candidates. An UV-exchange method allowed the generation of a repertoire of multimers including low-affinity HLA-binding peptides. These were labeled with quantum dot-fluorochromes and encoded in a combinatorial fashion, allowing parallel and sensitive detection of specific, low avidity T-cells. Significantly increased frequencies of T-cells against four novel PPI epitopes (PPI(4-13)/B7, PPI(29-38)/A2, PPI(76-84)/A3 and PPI(79-88)/A3) were detected in stored blood of patients with recent onset diabetes but not in controls. Changes in frequencies of circulating CD8 T-cells against these novel epitopes were detected in blood of islet graft recipients at different time points after transplantation, which correlated with clinical outcome. In conclusion, our novel strategy involving a sensitive multiplex detection technology and requiring minimal volumes of stored blood represents a major improvement in the direct ex-vivo characterization and enumeration of immune cells in the pathogenesis of type 1 diabetes. PMID- 21636248 TI - Complement and cytokine based therapeutic strategies in myasthenia gravis. AB - Myasthenia gravis (MG) is a T cell-dependent and antibody-mediated disease in which the target antigen is the skeletal muscle acetylcholine receptor (AChR). In the last few decades, several immunological factors involved in MG pathogenesis have been discovered mostly by studies utilizing the experimental autoimmune myasthenia gravis (EAMG) model. Nevertheless, MG patients are still treated with non-specific global immunosuppression that is associated with severe chronic side effects. Due to the high heterogeneity of AChR epitopes and antibody responses involved in MG pathogenesis, the specific treatment of MG symptoms have to be achieved by inhibiting the complement factors and cytokines involved in anti-AChR immunity. EAMG studies have clearly shown that inhibition of the classical and common complement pathways effectively and specifically diminish the neuromuscular junction destruction induced by anti-AChR antibodies. The inborn or acquired deficiencies of IL-6, TNF-alpha and TNF receptor functions are associated with the lowest EAMG incidences. Th17-type immunity has recently emerged as an important contributor of EAMG pathogenesis. Overall, these results suggest that inhibition of the complement cascade and the cytokine networks alone or in combination might aid in development of future treatment models that would reduce MG symptoms with highest efficacy and lowest side effect profile. PMID- 21636249 TI - T cell clonal expansions detected in patients with primary biliary cirrhosis express CX3CR1. AB - The intrahepatic biliary destruction of primary biliary cirrhosis (PBC) appears secondary to a multi-lineage response that includes autoantibodies, biliary apotopes, and cellular responses. Although there has been considerable effort in defining the role and specificity of anti-mitochondrial autoantibodies, a major challenge has been the characterization of T effector pathways. This difficulty is due in part to the limitation of current technologies for directly isolating and characterizing autoreactive T cells from patients. Herein, we successfully demonstrate a novel technology for characterizing the surface phenotype of T cell oligoclonal expansions directly ex vivo. Using PBC as a prototypic disease we were able to detect clonal T cell expansions in 15/15 patients examined. Although the T cell expansions from different patients expressed different TCRVbeta gene segments, the surface phenotype of the cells was the same. The clonal T cell expansions in PBC patients are CX3CR1(+) Fas(+) effector-memory T cells, a finding of particular importance given the known up-regulation of fractalkine on injured biliary epithelial cells (BEC). In contrast to the persistent aberrantly expanded T cells observed in the PBC patients, T cell expansions detected in response to a herpes viral infection were very dynamic and resolved over time. This protocol can be used to characterize T cell expansions in other autoimmune diseases. PMID- 21636251 TI - Effects of acetazolamide on epileptic apnea in migrating partial seizures in infancy. AB - We report two cases of migrating partial seizures in infancy complicated with intractable epileptic apnea with severe desaturation. Ictal electroencephalography revealed migrating foci of epileptiform discharges, which spread to bilateral temporal areas resulting in the onset of apnea. Magnetoencephalography detected dipole sources at bilateral perisylvian areas. Single photon emission tomography revealed a significant ictal change in perfusion at bilateral anterior temporal lobes in one patient. Addition of acetazolamide to the regimen resulted in complete disappearance of epileptic seizures. PMID- 21636252 TI - Abnormal movements are associated with poor psychosocial functioning in adolescents at high risk for psychosis. AB - The period immediately preceding the onset of overt psychosis is characterized by a range of symptoms and behaviors including emerging attenuated psychosis, spontaneous movement abnormalities, and a broad decline in role and social functioning. Recent evidence suggests that basal ganglia dysfunction, which is implicated in the development of psychotic symptomatology, may manifest in the form of both movement abnormalities and deficits in processes integral to psychosocial functioning. However, little is known about the relationship between abnormal movement function and the observed psychosocial deficits. In the present study, 40 clinical high-risk participants meeting criteria for a prodromal syndrome were assessed for movement abnormalities and global role and social functioning at baseline. Role and social functioning were then followed up after a one-year period. At baseline, the severity of spontaneous movement abnormalities was associated with poor role functioning. Further, when controlling for baseline functioning, movement abnormalities predicted changes in social functioning one-year later, with a trend in the same direction for role functioning. Exploratory analyses also indicated that elevated baseline movement abnormalities distinguished those at-risk participants who eventually converted to psychosis and that this was also the case for poorer baseline global role functioning (at the trend level). Taken together, the results suggest that movement abnormalities are closely associated with deficits in psychosocial functioning. Elucidating the link between these phenomena may serve to refine etiological models of frontal-subcortical circuit dysfunction and inform understanding of functioning and outcome of these affected youth. PMID- 21636253 TI - Translating genomic research into care for people with schizophrenia in China. AB - The implications of increased understanding of the genetic contribution to schizophrenia for patients and their families remain unclear. We carried out a study of Chinese patients'(n=118) and relatives' (n=78) views of illness severity, attribution of cause, concern about developing illness, and effect of schizophrenia on family planning. A comparison sample of English-survey respondents was also obtained, using the same series of questions (n=42 patients, n=127 relatives). Fewer Chinese patients and family members rated schizophrenia as very severe (33%) than did the predominantly North American respondents (67%, p<0.0001). The pattern of attribution of cause differed between samples (p<0.0001), favoring environmental alone in the Chinese sample (52%), with a low frequency of genetics alone (9%). Although comparatively fewer Chinese respondents were very concerned about developing schizophrenia themselves or about the risk of illness in their families (21%), this high level of concern was more common in family members (28%). Finally, Chinese respondents were somewhat less likely to indicate that schizophrenia impacted on family planning decisions (31%) than were English-survey respondents (45%, p=0.02). The descriptive findings contribute to understanding schizophrenia in China. The comparative findings must be regarded as preliminary, since differences in demographics could influence results. The present findings suggest that understanding patients' and families' attributions of cause of schizophrenia may be important for developing a shared model of illness in order to decrease stigmatization, and improve therapeutic alliances. PMID- 21636254 TI - Adherence to antipsychotic drug treatment in early-episode schizophrenia: a six month naturalistic follow-up study. AB - BACKGROUND: The Health Belief Model states that medication adherence is primarily determined by beliefs (i.e., perceptions of: adherence costs and benefits, susceptibility, and outcome severity), yet little is known regarding the model's longitudinal utility. AIMS: To examine the longitudinal utility of the Health Belief Model in explaining non-adherence with antipsychotic medication in clinical settings in early-episode schizophrenia. METHOD: Participants (n=112) with a DSM-IV diagnosis of schizophrenia (n=84, 75%) or schizoaffective disorder (n=28, 25%) participated in a four wave six month study. Participants were assessed on adherence (Visual Analog Scale for Assessing Treatment Adherence), symptom severity indices, the Drug Attitudes Inventory, and extrapyramidal side effects. RESULTS: Unlike non/partially adherent participants, adherent participants showed statistically significantly (p<.05) more: insight into illness, awareness of the need for medication, positive perceptions of trust in the doctor-patient therapeutic alliance, perceived family involvement in pharmacological treatment, positive attitudes towards medication in the family and fewer adverse events. Adherence rates at endpoint did not differ between typical, atypical and mixed antipsychotic medication groups. Structural equation modeling showed that over 6 months symptom severity, awareness for the need of medication and attitudes to medication predicted adherence. Awareness of the need of medication, awareness of social consequences, participant's perceived trust in physician and the severity of negative symptoms all predicted attitudes to medication that in turn predicted adherence. CONCLUSIONS: The current results partly support the adherence Health Belief Model, and emphasize the role of attitudes toward medication as a predictor of adherence. PMID- 21636255 TI - Vancomycin for meticillin-resistant Staphylococcus aureus (MRSA) infections in the Intensive Care Unit: the sunset boulevard of a gold standard? PMID- 21636256 TI - Vancomycin clearance during continuous venovenous haemofiltration in critically ill patients. AB - The objective of this study was to determine the pharmacokinetics and dosing recommendations of vancomycin in critically ill patients receiving continuous venovenous haemofiltration (CVVH). A prospective study was conducted in the Intensive Care Unit of a university hospital. Seven patients receiving CVVH with a triacetate hollow-fibre dialyser were enrolled. CVVH was performed in pre dilution mode with a blood flow rate of 200-250 mL/min and an ultrafiltrate flow rate of 800-1200 mL/h. To determine vancomycin pharmacokinetics, serum and ultrafiltrate were collected over 12 h after a 2-h infusion of 1000 mg vancomycin. The mean (+/- standard deviation) sieving coefficient of vancomycin was 0.71+/-0.13, which is consistent with previously reported values. Clearance of vancomycin by CVVH (0.73+/-0.21 L/h or 12.11+/-3.50 mL/min) constituted 49.4+/ 20.8% of total vancomycin clearance (1.59+/-0.47 L/h) and was consistent with previously reported clearances. Approximately one-fifth of the vancomycin dose was removed during the 12-h CVVH (213.9+/-104.0 mg). The volume of distribution was 24.69+/-11.00 L, which is smaller than previously reported. The elimination rate constant and terminal half-life were 0.08+/-0.05 h(-1) and 12.02+/-7.00 h, respectively. In conclusion, elimination of vancomycin by CVVH contributed to ca. 50% of the total elimination in critically ill patients. The maintenance dose of vancomycin, calculated from parameters from patients in this study, would be 500 750 mg every 12 h to provide a steady-state trough concentration of 15-20 mg/L. Owing to alterations in clinical conditions, serum vancomycin concentrations must be closely monitored in critically ill patients. PMID- 21636257 TI - Trial Simulation to estimate Type I error when a population window enrichment strategy is used to improve efficiency of clinical trials in depression. AB - Performance of recruitment centers is a critical feature for the success of multicentric randomized clinical trials (RCT). An enrichment window strategy (EWS) was recently proposed to enhance signal detection in RCTs based on the identification of centers with non-plausible placebo response, leading to a Per Protocol exclusion of all data from these non-informative centers before statistical analysis. The risk of an inflated Type I error associated with EWS was assessed using Clinical Trial Simulation. Randomized two-arm placebo controlled trials were simulated under the assumption that the drug treatment was ineffective.The results obtained in the absence of population enrichment were compared to the results obtained after EWS application. The results indicated that EWS preserved the Type I error showing no bias, at variance with other procedures. These results were confirmed by simulating 13 three-arm placebo controlled RCTs from the GSK Clinical Database. PMID- 21636258 TI - Regulation of endocytosis into human brain-microvascular endothelial cells by inhibition of efflux proteins. AB - The expressions of P-glycoprotein (P-gp) and multidrug resistance-associated proteins (MRPs) in the blood-brain barrier (BBB) were regulated by verapamil and probenecid. The two protein interveners inhibited the efflux-pumping capability for permeating calcein-AM in the monolayer of human brain-microvascular endothelial cells (HBMECs). The immunochemical staining revealed that the order in the integrity of tight junction was human astrocyte (HA)-regulated HBMECs>HBMECs cultured with 100% astrocyte-conditioned medium (ACM)>HBMECs cultured with 50% ACM>HBMECs. The viability of HBMECs was higher than 94% when the concentrations of verapamil and probenecid were lower than 50 MUM and 1000 MUM, respectively. The culture using ACM negligibly affected the activity of P-gp and MRPs on HBMECs after the suppression with verapamil and/or probenecid. However, the double culture of HA-regulated HBMECs promoted the quantity of P-gp and MRPs and reduced the endocytosis of calcein-AM. The inhibitive and endocytotic analysis can unveil the role of HAs in the protein expressions on HBMECs for establishing a reliable BBB model in vitro. PMID- 21636259 TI - Pain in patients with cancer: the World Health Organization analgesic ladder and beyond. PMID- 21636260 TI - Selection of thrombin-binding aptamers by using computational approach for aptasensor application. AB - The possibility of introducing a computationally assisted method to study aptamer protein interaction was evaluated with the aim of streamlining the screening and selection of new aptamers. Starting from information on the 15-mer (5' GGTTGGTGTGGTTGG-3') thrombin binding aptamer (TBA), a library of mutated DNA sequences (994 elements) was generated and screened using shapegauss a shape based scoring function from openeye software to generate computationally derived binding scores. The TBA and three other mutated oligonucleotides, selected on the basis of their binding score (best, medium, worst), were incorporated into surface plasmon resonance (SPR) biosensors. By reducing the ionic strength (binding buffer, 50 mM TrisHCl pH 7.4, 140 mM NaCl, 1mM MgCl2, diluted 1:50) in order to match the simulated condition, the analytical performances of the four oligonucleotide sequences were compared using signal amplitude, sensitivity (slope), linearity (R2) and reproducibility (CVav %). The experimental results were in agreement with the simulation findings. PMID- 21636261 TI - Reversible thrombin detection by aptamer functionalized STING sensors. AB - Signal Transduction by Ion NanoGating (STING) is a label-free technology based on functionalized quartz nanopipettes. The nanopipette pore can be decorated with a variety of recognition elements and the molecular interaction is transduced via a simple electrochemical system. A STING sensor can be easily and reproducibly fabricated and tailored at the bench starting from inexpensive quartz capillaries. The analytical application of this new biosensing platform, however, was limited due to the difficult correlation between the measured ionic current and the analyte concentration in solution. Here we show that STING sensors functionalized with aptamers allow the quantitative detection of thrombin. The binding of thrombin generates a signal that can be directly correlated to its concentration in the bulk solution. PMID- 21636262 TI - Micro/nanofabricated environments for synthetic biology. AB - A better understanding of how confinement, crowding and reduced dimensionality modulate reactivity and reaction dynamics will aid in the rational and systematic discovery of functionality in complex biological systems. Artificial microfabricated and nanofabricated structures have helped elucidate the effects of nanoscale spatial confinement and segregation on biological behavior, particularly when integrated with microfluidics, through precise control in both space and time of diffusible signals and binding interactions. Examples of nanostructured interfaces for synthetic biology include the development of cell like compartments for encapsulating biochemical reactions, nanostructured environments for fundamental studies of diffusion, molecular transport and biochemical reaction kinetics, and regulation of biomolecular interactions as functions of microfabricated and nanofabricated topological constraints. PMID- 21636263 TI - The role of diagnostic imaging in the evaluation of suspected osteomyelitis in the foot: a critical review. AB - The early diagnosis of osteomyelitis in the foot from its clinical presentation alone can be difficult particularly in cases when the early signs are subtle. Early diagnosis and subsequent early intervention are imperative to reduce the risk of chronic infection, associated early lytic changes to bone and potential long term structural complications caused by subsequent deformity and lost anatomy. Diagnostic imaging has a major role to play in the early assessment and diagnosis of bone infection, yet the choice of approach can be controversial. Several imaging modalities have been advocated, imaging of the infected foot is complex and no single test is ideal for every situation. The clinician needs to be aware of the strengths and weaknesses of each imaging modality so that the most appropriate test is selected for the individual case. Factors such as site of infection in the foot, the aggressive nature of the organism, the time since onset, previous associated surgery and co-morbidity may all play apart in the clinician's decision making process to determine the best approach in detecting the sometimes subtle changes which may be seen in some cases of osteomyelitis. This review considers the literature and highlights the advantages and disadvantages of the main imaging techniques used for the evaluation of the foot when osteomyelitis is suspected. An evidence based algorithm for the selection of appropriate imaging techniques is suggested to aid clinicians in there decision making process. PMID- 21636264 TI - Lower extremity radicular pain after prophylactic intrathecal saline injection through a subarachnoid catheter following accidental dural puncture. AB - We describe a case in which severe lower extremity radicular pain occurred after administration of 0.9% saline into the subarachnoid space through a catheter that had been left for 20 h following inadvertent dural puncture in an obstetric patient. A 42-year-old (G8P7) woman was admitted for repeat cesarean delivery. Accidental dural puncture occurred during epidural placement. Following a slow 10 mL intrathecal injection of 0.9% normal saline an epidural catheter was advanced into the subarachnoid space. Spinal anesthesia was used for cesarean delivery and the subarachnoid catheter was kept in place for 20 h. Before catheter removal, an additional 10 mL of 0.9% saline was slowly administered into the intrathecal space. Almost instantly, the patient complained of back pain that progressed to lower extremity radicular pain and paresthesia; symptoms began to resolve after 10 min. Subsequently, the patient developed a postdural puncture headache that persisted for three days. The patient's radiculitis and paresthesia likely resulted from an acute increase in intrathecal pressure after saline administration or from direct catheter irritation. Although both intrathecal saline administration and subarachnoid catheter placement have been previously proposed as ways to prevent postdural puncture headache, their efficacy remains controversial, and we advise caution with these techniques. PMID- 21636266 TI - Interneuron networks in the hippocampus. AB - The hippocampus has contributed enormously to our understanding of the operation of elemental brain circuits, not least through the classification of forebrain interneurons. Understanding the operation of interneuron networks however requires not only a wiring diagram that describes the innervation and postsynaptic targets of different GABAergic cells, but also an appreciation of the temporal dimension. Interneurons differ extensively in their intrinsic firing rates, their recruitment in different brain rhythms, and in their synaptic kinetics. Furthermore, in common with principal neurons, both the synapses innervating interneurons and the synapses made by these cells are highly modifiable, reflecting both their recent or remote use (short-term and long-term plasticity) and the action of extracellular messengers. This review examines recent progress in understanding how different hippocampal interneuron networks contribute to feedback and feed-forward inhibition at different timescales. PMID- 21636265 TI - From microRNAs to targets: pathway discovery in cell fate transitions. AB - MicroRNAs (miRNAs) are 22 nt non-coding RNAs that regulate expression of downstream targets by messenger RNA (mRNA) destabilization and translational inhibition. A large number of eukaryotic mRNAs are targeted by miRNAs, with many individual mRNAs being targeted by multiple miRNAs. Further, a single miRNA can target hundreds of mRNAs, making these small RNAs powerful regulators of cell fate decisions. Such regulation by miRNAs has been observed in the maintenance of the embryonic stem cell (ESC) cell cycle and during ESC differentiation. MiRNAs can also promote the dedifferentiation of somatic cells to induced pluripotent stem cells. During this process they target multiple downstream genes, which represent important nodes of key cellular processes. Here, we review these findings and discuss how miRNAs may be used as tools to discover novel pathways that are involved in cell fate transitions using dedifferentiation of somatic cells to induced pluripotent stem cells as a case study. PMID- 21636267 TI - Reduced miR-100 expression in cervical cancer and precursors and its carcinogenic effect through targeting PLK1 protein. AB - AIM: Although aberrant miRNAs expression has been documented, altered miR-100 expression in cervical cancer and precursor tissues and its carcinogenic effect and mechanism remain unexplored. The aim of our study was to investigate the role of miR-100 alteration in cervical carcinogenesis. METHODS: The expression of miR 100 was examined by quantitative real-time reverse transcriptase PCR (qRT-PCR) in 125 cervical tissues including normal cervical epithelium, cervical intraepithelial neoplasia (CIN), and cervical cancer, as well as in five cervical cell lines. Through modulating miR-100 expression using miR-100 inhibitor or mimic in vitro, cell growth, cycle and apoptosis were tested separately by MTT or flow cytometry and meanwhile Polo-like kinase1 (PLK1) mRNA and protein expressions were detected by qRT-PCR and immunoblotting. The expression of PLK1 in 125 cervical tissues was also examined by immunohistochemical staining and the correlation between miR-100 and PLK1 expression in the same tissues was analysed. Finally, HPV-16 E6/E7 expression was modulated by gene transfection and subsequently the level of miR-100 was examined by qRT-PCR. RESULTS: The miR-100 expression showed a significantly and gradually reduced tendency from low-grade CIN, high-grade CIN to cervical cancer tissues and a significant decrease in HPV positive cervical cancer cell lines. The modulation of miR-100 expression remarkably influenced cell proliferation, cycle and apoptosis, as well as the level of PLK1 protein, but not mRNA, in vitro experiments. PLK1 expression was negatively correlated with miR-100 expression in CIN3 and cervical cancer tissues. The modulation of HR-HPV E6/E7 expression did not change miR-100 level. CONCLUSIONS: The reduced miR-100 expression participates in the development of cervical cancer at least partly through loss of inhibition to target gene PLK1, which probably occurs in a relative late phase of carcinogenesis. HR-HPV E6/E7 may not directly regulate miR-100 expression in cervical cells. PMID- 21636268 TI - Laparoscopic hysterectomy is preferred over laparotomy in early endometrial cancer patients, however not cost effective in the very obese. AB - BACKGROUND: Total laparoscopic hysterectomy (TLH) is safe and cost effective in early stage endometrial cancer when compared to total abdominal hysterectomy (TAH). In non-randomised data it is often hypothesised that older and obese patients benefit most from TLH. Aim of this study is to analyse whether data support this assumption to advice patients, clinicians and policy makers. METHODS: Data of 283 patients enrolled in a randomised controlled trial comparing TAH versus TLH in early stage endometrial cancer were re-analysed. Randomisation by sequential number generation was done centrally, with stratification by trial centre. Using multivariate analysis, predictors of major complications and conversions to laparotomy were assessed. For the cost effectiveness analysis, subgroups of patients were constructed based on age and body mass index (BMI). For each subgroup, costs per major complication-free patient were estimated, using incremental cost effect ratios (extra costs per additional effect). RESULTS: Older (odds ratio (OR): 1.05; 1.01-1.09) and obese (OR: 1.05; 1.01-1.10) patients had a higher risk to develop complications, for both groups. In obese (OR: 1.17; 1.09-1.25) patients and patients with a previous laparotomy (OR: 3.45; 1.19-10.04) a higher risk of conversion to laparotomy was found. For patients>70 years of age and patients with a BMI over 35 kg/m2, incremental costs per major complication-free patients were ?16 and ?54 for TLH compared to TAH, respectively. CONCLUSION: In general, TLH should be recommended as the standard surgical procedure in early stage endometrial cancer, also in patients>70 years of age. In obese patients with a BMI>35 kg/m2 TLH is not cost effective because of the high conversion rate. A careful consideration of laparoscopic treatment is needed for this subgroup. Surgeon experience level may influence this choice. PMID- 21636269 TI - Thermo-mechanical extrusion pretreatment for conversion of soybean hulls to fermentable sugars. AB - Thermo-mechanical extrusion pretreatment for lignocellulosic biomass was investigated using soybean hulls as the substrate. The enzyme cocktail used to hydrolyze pretreated soybean hulls to fermentable sugars was optimized using response surface methodology (RSM). Structural changes in substrate and sugar yields from thermo-mechanical processing were compared with two traditional pretreatment methods that utilized dilute acid (1% sulfuric acid) and alkali (1% sodium hydroxide). Extrusion processing parameters (barrel temperature, in-barrel moisture, screw speed) and processing aids (starch, ethylene glycol) were studied with respect to reducing sugar and glucose yields. The conditions resulting in the highest cellulose to glucose conversion (95%) were screw speed 350rpm, maximum barrel temperature 80 degrees C and in-barrel moisture content 40%wb. Compared with untreated soybean hulls, glucose yield from enzymatic hydrolysis of soybean hulls increased by 69.6%, 128.7% and 132.2%, respectively, when pretreated with dilute acid, alkali and extrusion. PMID- 21636270 TI - Structure-based design of thienobenzoxepin inhibitors of PI3-kinase. AB - Starting from thienobenzopyran HTS hit 1, co-crystallization, molecular modeling and metabolic analysis were used to design potent and metabolically stable inhibitors of PI3-kinase. Compound 15 demonstrated PI3K pathway suppression in a mouse MCF7 xenograft model. PMID- 21636271 TI - Enantioselective binding of second generation pyrrolobenzoxazepinones to the catalytic ternary complex of HIV-1 RT wild-type and L100I and K103N drug resistant mutants. AB - We investigated some pyrrolobenzoxazepinone (PBOs, 3e-i) analogues of early described effective non-nucleoside inhibitors of HIV-1 reverse transcriptase (RT). Enzymological studies of 3e-i enantiomers, with wild type (wt) RT and some drug-resistant mutants, revealed a stereoselective mode of action and selectivity for RT ternary complex. Unexpectedly (+)-3g was found more potent towards the L100I mutant than towards the wt RT, whereas (+)-3h inhibited the K103N mutant and RT wt with comparable potency. PMID- 21636272 TI - The geranyl-modified tryptophan residue is crucial for ComXRO-E-2 pheromone biological activity. AB - The ComX pheromone is an isoprenoidal oligopeptide containing a modified tryptophan residue, which stimulates natural genetic competence in gram-positive bacteria, Bacillus. We have reported the structure of the ComX(RO-E-2) pheromone, which is produced by the RO-E-2 strain of Bacillus subtilis. ComX(RO-E-2) analogs with substituted amino acids and isoprenoid modified tryptophan residues (e.g., prenyl, geranyl, and farnesyl), were synthesized and examined for biological activity. These results indicate that Phe-Trp(*)(Ger)-NH(2) is the minimum pharmacophore of the ComX(RO-E-2) pheromone. Furthermore, the length of the isoprenoid moiety (i.e., modification style), and the presence of double bonds, are crucial for biological activity. The modification style of the ComX pheromone is more important than the peptide sequence with respect to biological activity. PMID- 21636273 TI - Substituted benzoxazinones as potent positive allosteric AMPA receptor modulators: part II. AB - AMPA receptors (AMPARs) are an important therapeutic target in the CNS. A series of substituted benzoxazinone derivatives with good to very good in vitro activity as positive allosteric AMPAR modulators was synthesized and evaluated. The appropriate substituent choice on the benzoxazinone fragment improved the affinity towards the AMPA receptor significantly in comparison to our lead molecule CX614. PMID- 21636274 TI - Syntheses of 2-NBDG analogues for monitoring stereoselective uptake of D-glucose. AB - 2-NBDG is a widely used fluorescent tracer for monitoring d-glucose uptake into single living cells. However, 2-NBDG alone is not sufficient for monitoring the net stereoselective uptake of d-glucose, unless its possible non-stereoselective uptake is properly evaluated. l-Glucose derivatives, which emit fluorescence distinct from that of 2-NBDG, should provide valuable information on the stereoselective uptake, when used with 2-NBDG in combination. In the present study, we synthesized Texas Red (sulforhodamine 101 acid)-coupled and [2-(benz-2 oxa-1,3-diazol-4-yl)amino]-coupled 2-deoxy-D-glucose, referred to as [2-TRG] and [2-BDG], respectively. These derivatives showed emission wavelength longer and shorter than that of 2-NBDG, respectively. 2-TRLG, an antipode of 2-TRG, proved to be an effective tracer for evaluating the extent of non-stereoselective uptake of 2-NBDG when used simultaneously with 2-NBDG. On the other hand, 2-BDG exhibited very weak fluorescence, but the application of a novel cross coupling in the presence of a benzoxadiazole group may be useful for the future development of effective glucose tracers. PMID- 21636275 TI - Benzoxazinones as potent positive allosteric AMPA receptor modulators: part I. AB - AMPA receptors (AMPARs) are an increasingly important therapeutic target in the CNS. Aniracetam, the first identified potentiator of AMPARs, led to the rigid and more potent CX614. This lead molecule was optimized in order to increase affinity towards the AMPA receptor. The substitution of the dioxine with a benzoxazinone ring system increased the activity and allowed further investigation of the sidechain SAR. PMID- 21636276 TI - Pyridazine-derived gamma-secretase modulators. AB - SAR of a novel series of pyridazine-derived gamma-secretase modulators is described. Compound 25 was found to be a potent modulator in vitro, which on further profiling, was found to decrease Abeta42 and Abeta40, and maintain the levels of total Abeta. Furthermore, 25 demonstrated excellent pharmacokinetic parameters as well as good CNS penetration in the rat. PMID- 21636277 TI - Agitated honeybees exhibit pessimistic cognitive biases. AB - Whether animals experience human-like emotions is controversial and of immense societal concern [1-3]. Because animals cannot provide subjective reports of how they feel, emotional state can only be inferred using physiological, cognitive, and behavioral measures [4-8]. In humans, negative feelings are reliably correlated with pessimistic cognitive biases, defined as the increased expectation of bad outcomes [9-11]. Recently, mammals [12-16] and birds [17-20] with poor welfare have also been found to display pessimistic-like decision making, but cognitive biases have not thus far been explored in invertebrates. Here, we ask whether honeybees display a pessimistic cognitive bias when they are subjected to an anxiety-like state induced by vigorous shaking designed to simulate a predatory attack. We show for the first time that agitated bees are more likely to classify ambiguous stimuli as predicting punishment. Shaken bees also have lower levels of hemolymph dopamine, octopamine, and serotonin. In demonstrating state-dependent modulation of categorization in bees, and thereby a cognitive component of emotion, we show that the bees' response to a negatively valenced event has more in common with that of vertebrates than previously thought. This finding reinforces the use of cognitive bias as a measure of negative emotional states across species and suggests that honeybees could be regarded as exhibiting emotions. PMID- 21636278 TI - Pervasive multinucleotide mutational events in eukaryotes. AB - Many aspects of mutational processes are nonrandom, from the preponderance of transitions relative to transversions to the higher rate of mutation at CpG dinucleotides [1]. However, it is still often assumed that single-nucleotide mutations are independent of one another, each being caused by separate mutational events. The occurrence of multiple, closely spaced substitutions appears to violate assumptions of independence and is often interpreted as evidence for the action of adaptive natural selection [2, 3], balancing selection [4], or compensatory evolution [5, 6]. Here we provide evidence of a frequent, widespread multinucleotide mutational process active throughout eukaryotes. Genomic data from mutation-accumulation experiments, parent-offspring trios, and human polymorphisms all show that simultaneous nucleotide substitutions occur within short stretches of DNA. Regardless of species, such multinucleotide mutations (MNMs) consistently comprise ~3% of the total number of nucleotide substitutions. These results imply that previous adaptive interpretations of multiple, closely spaced substitutions may have been unwarranted and that MNMs must be considered when interpreting sequence data. PMID- 21636279 TI - The kynurenine pathway modulates neurodegeneration in a Drosophila model of Huntington's disease. AB - Neuroactive metabolites of the kynurenine pathway (KP) of tryptophan degradation have been implicated in the pathophysiology of neurodegenerative disorders, including Huntington's disease (HD) [1]. A central hallmark of HD is neurodegeneration caused by a polyglutamine expansion in the huntingtin (htt) protein [2]. Here we exploit a transgenic Drosophila melanogaster model of HD to interrogate the therapeutic potential of KP manipulation. We observe that genetic and pharmacological inhibition of kynurenine 3-monooxygenase (KMO) increases levels of the neuroprotective metabolite kynurenic acid (KYNA) relative to the neurotoxic metabolite 3-hydroxykynurenine (3-HK) and ameliorates neurodegeneration. We also find that genetic inhibition of tryptophan 2,3 dioxygenase (TDO), the first and rate-limiting step in the pathway, leads to a similar neuroprotective shift toward KYNA synthesis. Importantly, we demonstrate that the feeding of KYNA and 3-HK to HD model flies directly modulates neurodegeneration, underscoring the causative nature of these metabolites. This study provides the first genetic evidence that inhibition of KMO and TDO activity protects against neurodegenerative disease in an animal model, indicating that strategies targeted at two key points within the KP may have therapeutic relevance in HD, and possibly other neurodegenerative disorders. PMID- 21636280 TI - Total knee arthroplasty for secondary osteoarthritis following ACL reconstruction: a matched-pair comparative study of intra-operative and early post-operative complications. AB - Injury to the anterior cruciate ligament (ACL) is associated with increased risk of osteoarthritis and subsequent need for total knee arthroplasty (TKA). The impact of prior ACL reconstruction on TKA has been rarely studied. Twenty-two patients undergoing TKA, with a mean of 26 years following ACL reconstruction, were compared to a matched control group. Tibial exposure was more difficult in the study group, requiring tibial tubercle osteotomy in three cases (14%). Manipulation under anesthesia was required in five patients in the study group (23%) and none in the control group (p=0.048). No differences in final range of motion, outcomes scores, or alignment were noted 2 to 3 year post-operative. Total knee arthroplasty following ACL reconstruction is effective. Difficulties in obtaining tibial exposure and post-operative stiffness requiring manipulation under anesthesia are common. PMID- 21636281 TI - Revision of failed unicompartmental knee replacement to total knee replacement. AB - Recent reports suggest good outcome results following unicompartmental knee replacement (UKR). However, a number of authors have commented on the problem of osseous defects requiring technically difficult revision surgery. We reviewed clinical outcomes following revision total knee replacement (TKR) for failed UKR and analysed the reasons for failure and the technical aspects of the revision surgery. Between 2001 and 2010 our institute performed 132 UKR's out of which 33 required revision to TKR during a period 6 years. Demographics, details and indications for primary and revision surgery, the revised prosthesis including augments, technical difficulties and complications were noted. Patient outcome assessment was based on the Oxford knee score (OKS). Survival analysis for the UKR prosthesis was calculated using Kaplan-Meier Survival curves. Reasons for revision included aseptic loosening, persistent pain, dislocated meniscus, mal alignment and other compartment osteoarthritis. Median time to revision was 19 months (range 2-159). Using revision as the end-point the survival proportion at 5-years was 69%. 18 revisions required additional intra-operative constructs including stemmed implants, wedge augmentation or bone graft. The mean 1 year post-operative OKS was 29 compared to 39 for primary TKR during the same period (p<0.001). Aseptic loosening was the commonest mode of failure. UKR survivorship at a non-specialist institute is considerably lower than at originating centres. Two thirds of the revisions were technically difficult and required additional constructs. The clinical outcome after revision surgery was inferior to that of primary TKR. The role of UKR needs to be more clearly defined. PMID- 21636282 TI - A hydrophilic polymer grafted with a histone tail peptide as an artificial gene regulator. AB - In chromatin, gene transcription is regulated through posttranslational modifications on the histone N-terminal tail sequences, typically an acetyl group modification on lysine residues. To realize a simple model of the gene regulation of chromatin, we designed a hydrophilic polymer grafted with histone H3 tail peptides. The polyplex formed from the polymer and DNA suppressed the gene expression effectively although the polyplex was weaker than the polyplex of poly L-lysine and DNA. This weaker polyplex afforded the acetylation of the lysine residue of the grafted peptides by histone acetyltransferase. Subsequently, the gene expression was activated due to the relaxation of the polyplex which was brought by a cationic charge decrease in the grafted peptides. This molecular system is the first functional model of the gene regulation of the chromatin. PMID- 21636283 TI - Influence of mineralogical and heavy metal composition on natural radionuclide concentrations in the river sediments. AB - The natural radiation level has been determined for the sediment samples of the Ponnaiyar River with an aim of evaluating the radiation hazard. The mineralogical characterizations of the sediments have been carried out using the Fourier Transform Infrared (FTIR) spectroscopic technique. The relative distribution of major minerals is determined by calculating extinction coefficient. The concentration and spatial distribution of heavy metals (Pb, Cr, Cu, Zn and Ni) have been studied to understand the heavy metal contamination and its level of toxicity. To evaluate the potential toxicity, heavy metal concentrations are compared with different toxicological and geological reference values. The comparison results suggest that the present metals create an adverse effect on the aquatic ecosystems associated with this river. To assess the sediment contamination due to the studied heavy metals, the Pollution Load Index (PLI) is calculated. Multivariate Statistical analyses (Pearson Correlation, Cluster and Factor analysis) were carried out between the parameters obtained from radioactivity, mineralogical and geochemical analysis to know the existing relations. Obtained results showed that the effect of mineralogy on level of radioactivity should be significant. However, mineralogy effect on heavy metal composition in the sediments should be limited, indicating that other factors such as vicinity of the pollution sources are more important. Also, the influence of mineralogical characterization on level of radioactivity is significant, whereas the influence of the heavy metal composition on level of radioactivity should be limited. PMID- 21636284 TI - Ascending aortic approach for balloon aortic valvuloplasty with concomitant bilateral pulmonary artery banding in a very low-birth-weight neonate with critical aortic stenosis and poor left ventricular function. AB - An 1186-g infant was born at 27 weeks' gestation. Echocardiography showed critical aortic stenosis, a dilated left ventricle (left ventricular end diastolic diameter, 16.9 mm), and poor left ventricular function (left ventricular ejection fraction, 8.5%). Due to duct-dependent systemic circulation, the patient underwent a hybrid intervention consisting of bilateral pulmonary artery banding (PAB) and balloon aortic valvuloplasty (BAV) via the ascending aorta on day 2. A stainless steel tip and a 6-French introducer were combined to create an access device. The access device was inserted into the ascending aorta, through which balloon catheters were manipulated. On day 4, echocardiography showed a left ventricular ejection fraction of 48% and dominant antegrade blood flow in the aortic arch. The pulmonary artery bands were removed and the ductus was clipped on day 8 to establish in-line circulation. After re-balloon aortic valvuloplasty for restenosis, the patient was discharged from the hospital at 7 months of age. The clinical implications of this case are: the ascending aortic approach is feasible for BAV in low-birth-weight neonates; and bilateral PAB performed concomitantly with BAV may be efficient for neonates with critical aortic stenosis and poor left ventricular function. PMID- 21636285 TI - Hyperoxia during early reperfusion does not increase ischemia/reperfusion injury. AB - OBJECTIVE: Oxygen is routinely administered to patients undergoing acute myocardial infarction as well as during revascularization procedures and cardiac surgery. Because reactive oxygen species are mediators of ischemia/reperfusion injury, increased oxygen availability might theoretically aggravate myocardial injury during reperfusion. We hypothesized that ventilation with a hyperoxic gas at start of reperfusion might increase ischemia/reperfusion injury. METHODS: Rats were anesthetized with isoflurane and ventilated with 40% oxygen. The animals were subjected to 40 min of regional myocardial ischemia and 120 min of reperfusion. In the test group, rats (n=11) were ventilated with a normobaric hyperoxic gas (95% O2) during the last 10 min of ischemia and the first 10 min of reperfusion. Control rats (n=14) were ventilated with 40% O2 throughout the experiments. Due to irreversible reperfusion arrhythmias, one animal in the hyperoxia group and six animals in the control group were excluded. Hearts (n=8 in the control group and n=10 in the test group) were harvested for measurement of infarct size. RESULTS: The incidence of lethal arrhythmias was 1/11 in the test group and 6/14 in the control group (p=0.06). Reperfusion with normobaric hyperoxia did not influence infarct size (20+/-8% of area at risk) compared with the normoxia group (24+/-8% and of area at risk), respectively (mean+/-SD, p>0.2). CONCLUSION: Normobaric hyperoxia during early reperfusion did not increase ischemia/reperfusion injury. PMID- 21636286 TI - Thrombus arising from ascending aorta. PMID- 21636287 TI - Aspirin resistance in off-pump coronary artery bypass grafting. AB - OBJECTIVE: Anti-platelet therapy with aspirin is the cornerstone of treatment after coronary artery bypass grafting (CABG). Aspirin resistance describes the clinical observation of the inability of aspirin to prevent thrombotic complications or the laboratory phenomenon of absence of the effect of aspirin on platelet inhibition tests. Off-pump CABG (OPCAB) is associated with reduced platelet activation and turnover compared to on-pump surgery which may indicate that aspirin is more effective after OPCAB. Our aim was to evaluate the efficacy of aspirin and incidence of aspirin resistance in patients undergoing OPCAB. METHODS: A total of 331 patients was recruited, of which 111 underwent primary OPCAB (group A) and 220 controls with ischaemic heart disease received medical therapy. Arachidonic acid-induced platelet aggregation and urinary 11-dehydro thromboxane B2 (11-dehydroTxB2) were measured at baseline and following aspirin administration on days 1, 4 and 10. A 6-month follow-up was completed in patients who developed aspirin resistance. RESULTS: On the first postoperative day, 78 patients (70.3%) were aspirin sensitive (AS) and 33 (29.7%) were aspirin resistant (AR). Of the latter, 18 (16.2%) and five (4.5%) patients remained resistant on days 4 and 10, respectively. AR patients had significantly greater platelet aggregation and urinary 11-dehydroTxB2 levels at all time points than those in the AS group. All patients in the AR group were AS by 6 months. All controls were sensitive to aspirin with similar platelet aggregation and 11 dehydroTxB2 to those in the AS group. CONCLUSIONS: Aspirin resistance is a transient phenomenon during the early postoperative period in approximately 30% of patients undergoing OPCAB. PMID- 21636288 TI - Vitamin D status and its associated factors of free living Malay adults in a tropical country, Malaysia. AB - Vitamin D status is influenced by sun exposure, geographic latitude, daily outdoor activities, body surface exposed to sunlight and dietary intakes. Malaysia, is sunny all year round. However, the vitamin D status of this population especially among the healthy and free living adults is not known. Therefore a study of vitamin D status and associated factors was initiated among an existing Malay cohort in Kuala Lumpur. A total of 380 subjects were sampled to have their vitamin D status assessed using 25-hydroxyvitamin D (25(OH)D). A short questionnaire enquiring socio-demographic characteristics, exposure to sunlight and clothing style was administered. Their mean age was 48.5+/-5.2years and the mean 25(OH)D for males and females were 56.2+/-18.9nmol/L and 36.2+/-13.4nmol/L respectively. There were significant positive correlation for sun exposure score (r=0.27, p<0.001) and negative correlation for sun protection score (r=-0.41, p<0.001) with 25(OH)D levels. In the logistic regression model, females (OR=2.93; 95% CI: 1.17, 7.31), BMI (1.1; 1.03, 1.20) and sun exposure score (0.998; 0.996, 0.999) were significantly associated with vitamin D status as represented by 25(OH)D levels. Our findings show that obesity, lifestyle behaviours and clothing style are directly associated with our participants especially females' low vitamin D status. PMID- 21636289 TI - Systematic review of efficacy of dose-dense versus non-dose-dense chemotherapy in breast cancer, non-Hodgkin lymphoma, and non-small cell lung cancer. AB - Randomized controlled trials (RCTs) have suggested a potential advantage of dose dense chemotherapy in improving disease-free and overall survival in patients with certain malignancies. This systematic review summarizes the literature on the efficacy of dose-dense chemotherapy across various cancers (breast cancer, non-Hodgkin lymphoma [NHL], and non-small cell lung cancer) and chemotherapy regimens. Among the 17 trials identified, few reported statistically significant differences between dose-dense and standard chemotherapy, and most were small with limited statistical power. Statistically significant differences in overall survival favoring dose-dense schedules were apparent among large RCTs in potentially curative settings such as early-stage breast cancer and NHL. Clinical and treatment heterogeneity demonstrated the flexibility of the dose-dense paradigm but also precluded quantitative meta-analysis of results. Further study of dose-dense schedules based on large RCTs is needed to demonstrate the consistency and generalizability of these findings. PMID- 21636290 TI - The effect of the partially restricted sit-to-stand task on biomechanical variables in subjects with and without Parkinson's disease. AB - The aim of this study was to explore the electromyographic, kinetic and kinematic patterns during a partially restricted sit-to-stand task in subjects with and without Parkinson's disease (PD). If the trunk is partially restricted, different behavior of torques and muscle activities could be found and it can serve as a reference of the deterioration in the motor performance of subjects with PD. Fifteen subjects participated in this study and electromyography (EMG) activity of the tibialis anterior (TA), soleus (SO), vastus medialis oblique (VMO), biceps femoris (BF) and erector spinae (ES) were recorded and biomechanical variables were calculated during four phases of the movement. Subjects with PD showed more flexion at the ankle, knee and hip joints and increased knee and hip joint torques in comparison to healthy subjects in the final position. However, these joint torques can be explained by the differences in kinematic data. Also, the hip, knee and ankle joint torques were not different in the acceleration phase of movement. The use of a partially restricted sit-to-stand task in PD subjects with moderate involvement leads to the generation of joint torques similar to healthy subjects. This may have important implications for rehabilitation training in PD subjects. PMID- 21636291 TI - Concepts or metacognition--what is the issue? Commentary on Stephane Savanah's "The concept possession hypothesis of self-consciousness". AB - The author claims that concept possession is not only necessary but also sufficient for self-consciousness, where self-consciousness is understood as the awareness of oneself as a self. Further, he links concept possession to intelligent behavior. His ultimate aim is to provide a framework for the study of self-consciousness in infants and non-human animals. I argue that the claim that all concepts are necessarily related to the self-concept remains unconvincing and suggest that what might be at issue here are not so much conceptual but rather metacognitive abilities. PMID- 21636292 TI - Osteochondritis dissecans of the first metatarsophalangeal joint: arthroscopy and microfracture technique. AB - Osteochondritis dissecans (OCD) is a localized disorder of the joint surface by which a segment of subchondral bone and its overlying cartilage become fragmented. Arthroscopic treatment of OCD of the first metatarsophalangeal joint consisted mainly of debridement and loose body removal. The microfracture technique for the treatment of OCD of the first metatarsophalangeal joint is presented for the first time. The technique has been shown to be a safe and technically uncomplicated first-line treatment. PMID- 21636293 TI - Functional outcomes after fibula locking nail for fragility fractures of the ankle. AB - The aim of the present study was to assess the functional outcome of fragility fractures of the ankle treated with a fibular locking nail. A retrospective review of 24 patients with fragility fractures treated with a fibular locking nail from January 2005 to December 2007 was performed. The fibular nail used in our study was Biomet SST (stainless steel taper) small bone locking nail for the fibula. The Olerud and Molander scale was used to assess the functional outcome at the end of 1 year. The domains of the Olerud and Molander scale are pain, stiffness, swelling, stair climbing, running, jumping, squatting, support, and the activities of daily living. The patients were interviewed by telephone or the questionnaire was send by mail. Of the 24 patients, 2 were men and 22 were women. The left side was affected in 15 patients. The age group ranged from 71 to 91 years (average, 79). Of the fractures, 10 were lateral alveolus, 8 were bimalleolar, and 6 were trimalleolar fractures. All the patients were followed up at 6 weeks, 12 weeks, and after 6 months. The average period to fracture union was 8.7 weeks. No wound breakdown or any deep infections developed. The average Olerud and Molander scale score was 57 (range 30 to 65). The use of fibular locking nails to treat these difficult fracture are quite crucial to achieve early mobilization and also to maintain a good fracture position. In our study, the use of fibular nails was a very useful and successful method of treating fragility fractures with a very low risk of complications. It also helps to restore function and results in patient satisfaction. PMID- 21636294 TI - Bcl6 protein expression shapes pre-germinal center B cell dynamics and follicular helper T cell heterogeneity. AB - The transcription factor Bcl6 is essential for the development of germinal center (GC) B cells and follicular helper T (Tfh) cells. However, little is known about in vivo dynamics of Bcl6 protein expression during and after development of these cells. By using a Bcl6 reporter mouse strain, we found that antigen-engaged B cells upregulated Bcl6 before clustering in GCs. Two-photon microscopic analysis indicated that Bcl6 upregulation in pre-GC B cells contributed to sustaining their interactions with helper T cells and was required for their entry to GC clusters. Our data also suggested that Tfh cells gradually downmodulated Bcl6 protein over weeks after development. The Bcl6-low Tfh cells rapidly terminated proliferation and upregulated IL-7 receptor. These results clarify the role of Bcl6 in pre-GC B cell dynamics and highlight the modulation of Bcl6 expression in Tfh cells that persist in the late phase of the antibody response. PMID- 21636295 TI - Germinal center B cell and T follicular helper cell development initiates in the interfollicular zone. AB - We identify the interfollicular (IF) zone as the site where germinal center B cell and T follicular helper (Tfh) cell differentiation initiates. For the first 2 days postimmunization, antigen-specific T and B cells remained confined within the IF zone, formed long-lived interactions, and upregulated the transcriptional repressor Bcl6. T cells also acquired the Tfh cell markers CXCR5, PD-1, and GL7. Responding B and T cells migrated to the follicle interior directly from the IF zone, T cell immigration preceding B cells by 1 day. Notably, in the absence of cognate B cells, Tfh cells still formed and migrated to the follicle. However, without such B cells, PD-1, ICOS, and GL7 were no longer expressed on follicular Bcl6(hi) T cells that nevertheless persisted in the follicle. Thus, Ag-specific B cells are required for the maintenance of the PD-1(hi)ICOS(hi)GL7(hi) Tfh cell phenotype within the follicle, but not for their initial differentiation in the IF zone. PMID- 21636297 TI - Insomnia and daytime cognitive performance: a meta-analysis. AB - OBJECTIVES: Individuals with insomnia consistently report difficulties pertaining to their cognitive functioning (e.g., memory, concentration). However, objective measurements of their performance on neuropsychological tests have produced inconsistent findings. This meta-analysis was conducted to provide a quantitative summary of evidence regarding the magnitude of differences between individuals with primary insomnia and normal sleepers on a broad range of neuropsychological measures. METHODS: Reference databases (PubMed, PsycInfo, Dissertation Abstracts International) were searched for studies comparing adults with primary insomnia to normal sleepers on neuropsychological measures. Dependent variables related to cognitive and psychomotor performance were extracted from each study. Variables were classified independently by two licensed neuropsychologists according to the main cognitive function being measured. Individual effect sizes (Cohen's d) were weighted by variability and combined for each cognitive function using a fixed effects model. Average effect sizes and their 95% confidence intervals were computed for each cognitive function. RESULTS: Twenty-four studies met inclusion criteria, for a total of 639 individuals with insomnia and 558 normal sleepers. Significant impairments (p<0.05) of small to moderate magnitude were found in individuals with insomnia for tasks assessing episodic memory (ES = -0.51), problem solving (ES = -0.42), manipulation in working memory (ES = -0.42), and retention in working memory (ES = -0.22). No significant group differences were observed for tasks assessing general cognitive function, perceptual and psychomotor processes, procedural learning, verbal functions, different dimensions of attention (alertness, complex reaction time, speed of information processing, selective attention, sustained attention/vigilance) and some aspects of executive functioning (verbal fluency, cognitive flexibility). CONCLUSION: Individuals with insomnia exhibit performance impairments for several cognitive functions, including working memory, episodic memory and some aspects of executive functioning. While the data suggests that these impairments are of small to moderate magnitude, further research using more ecologically valid measures and normative data are warranted to establish their clinical significance. PMID- 21636296 TI - ICOS receptor instructs T follicular helper cell versus effector cell differentiation via induction of the transcriptional repressor Bcl6. AB - The nature of follicular helper CD4(+) T (Tfh) cell differentiation remains controversial, including the minimal signals required for Tfh cell differentiation and the time at which Tfh cell differentiation occurs. Here we determine that Tfh cell development initiates immediately during dendritic cell (DC) priming in vivo. We demonstrate that inducible costimulator (ICOS) provides a critical early signal to induce the transcription factor Bcl6, and Bcl6 then induces CXCR5, the canonical feature of Tfh cells. Strikingly, a bifurcation between Tfh and effector Th cells was measurable by the second cell division of CD4(+) T cells, at day 2 after an acute viral infection: IL2Ralpha(int) cells expressed Bcl6 and CXCR5 (Tfh cell program), whereas IL2Ralpha(hi) cells exhibited strong Blimp1 expression that repressed Bcl6 (effector Th cell program). Virtually complete polarization between Bcl6(+) Tfh cells and Blimp1(+) effector Th cell populations developed by 72 hr, even without B cells. Tfh cells were subsequently lost in the absence of B cells, demonstrating a B cell requirement for maintenance of Bcl6 and Tfh cell commitment via sequential ICOS signals. PMID- 21636298 TI - Effects of medetomidine and ketamine on the regional cerebral blood flow in cats: a SPECT study. AB - Brain perfusion can be investigated using single photon emission computed tomography (SPECT) and the intravenous injection of (99m)technetium ethyl cysteinate dimer ((99m)Tc-ECD). However, sedation using medetomidine, an alpha(2) agonist, or anaesthesia using medetomidine and ketamine, an N-methyl-d-aspartate (NMDA)-antagonist, may be required for SPECT studies in cats but can affect the regional cerebral blood flow (rCBF). The effects of medetomidine, with or without ketamine, on regional brain perfusion were therefore investigated in six cats under three conditions. Injection of tracer occurred before sedation or anaesthesia (condition A), following intramuscular (IM) sedation with medetomidine (condition M) or after IM anaesthesia with medetomidine and ketamine (condition MK). Medetomidine and medetomidine with ketamine caused a significantly higher total tracer uptake in all brain regions. Semi quantification of brain perfusion gave lower perfusion indices in several sub cortical regions in conditions M and MK, compared to A. Left-right differences were observed in the temporal cortex (A), the temporal, parietal cortex and the thalamus (M) and the frontal cortex (MK). A significantly higher perfusion index in the sub-cortical regions, compared to the whole cortex, was only present in condition A. This study showed that caution is needed when quantifying brain perfusion indices when using sedative or anaesthetic agents that may affect rCBF. PMID- 21636299 TI - IGF-I-IGFBP-3-acid-labile subunit (ALS) complex in children and adolescents with classical congenital adrenal hyperplasia due to 21-hydroxylase deficiency (CAH). AB - It has been shown that changes in IGF-I and IGFBP levels in children with classical congenital adrenal hyperplasia due to 21-hydroxylase deficiency (CAH) are correlated with different states of metabolic control. Our approach was to analyze the serum levels of IGF-I, IGFBP-3, their molar ratio IGF-I:IGFBP-3 (MR), and ALS in a cohort of CAH children and adolescents, and their associations with different clinical and biochemical parameters. DESIGN AND PATIENTS: 56 patients, aged between 5.6 and 19.0 years were studied cross-sectionally. All patients had genetically proven CAH and received standard steroid substitution therapy. We measured serum levels of IGF-I, IGFBP-3, and ALS by commercial ELISA and calculated MR and assigned population-based SD scores (SDS). RESULTS: (median, quartiles) Overall IGF-I was not significantly altered (0.05 SDS, -1.21, 0.92), whereas IGFBP-3 was significantly elevated (1.50 SDS; 0.58, 1.95, p<0.0001) compared to the reference population. Consecutively, MR was decreased (-0.64 SDS; -1.38, 0.32; p=0.0017). ALS was clearly decreased (-1.95 SDS; -3.075, -1.00; p<0.0001). ALS, IGF-I, MR, and IGFBP-3 SDS were lower in pubertal than in prepubertal patients (p<0.05). ALS SDS were lower in girls (p=0.0038). Correlation analyses (r(s), p) revealed correlations between MR/ALS and chronological age (-0.583, <0.0001/-0.428, 0.0010), MR/ALS and Tanner stages ( 0.500, <0.0001/-0.334, 0.0118), MR/ALS and bone age (0.407, 0.0075/0.426, 0.0049), and between MR and ALS (0.405, 0.0020), respectively. For MR and ALS, we found no significant correlations for BMI, HOMA-IR, hydrocortisone and fludrocortisone dosage, or parameters of metabolic control. CONCLUSIONS: Our data provide evidence that the components of the trimeric IGF-I-IGFBP-3-ALS complex are altered in CAH children with possible implications on pubertal growth and final height. PMID- 21636300 TI - A large TAT deletion in a tyrosinaemia type II patient. AB - A girl, born to unrelated Spanish parents, presented at 6 months of age with photophobia, keratitis, palmar hyperkeratosis and high plasma tyrosine levels, indicative of tyrosinaemia type II. Analysis of the tyrosine aminotransferase (TAT) gene revealed a paternally inherited frameshift mutation c.1213delCinsAG at codon 405 causing a premature stop codon, and a maternally inherited deletion of 193kb encompassing the complete TAT gene and three neighbouring genes. This is the first complete TAT deletion in tyrosinaemia type II described so far. PMID- 21636301 TI - Time-dependent changes in the plasma amino acid concentration in diabetes mellitus. AB - We investigated longitudinal change in the amino acid (AA) profile in type 1 diabetes mellitus (DM) using AKITA mice, which develop DM as a result of insulin deficiency. The plasma concentrations of valine, leucine, isoleucine, as well as the total branched chain amino acids, alanine, citrulline and proline, were significantly higher in the diabetic mice. We show that the degree and timing of the changes were different among the plasma amino acid concentrations (pAAs) during the development of type 1 DM. PMID- 21636302 TI - Early-onset severe neuromuscular phenotype associated with compound heterozygosity for OPA1 mutations. AB - INTRODUCTION: Pathogenic mutations in the OPA1 gene are the most common identifiable cause of autosomal dominant optic atrophy (DOA), which is characterized by selective retinal ganglion cell loss, a distinctive pattern of temporal pallor of the optic nerve and a typical color vision deficit, with variable effects on visual acuity. Haploinsufficiency has been suggested as the major pathogenic mechanism for DOA. Here we present two siblings with severe ataxia, hypotonia, gastrointestinal dysmotility, dysphagia, and severe, early onset optic atrophy who were found to be compound heterozygotes for two pathogenic OPA1 mutations. This example expands the clinical phenotype of OPA1 associated disorders and provides additional evidence for semi-dominant inheritance. METHODS AND RESULTS: Molecular analysis of the OPA1 gene in this family by Sanger sequencing revealed compound heterozygosity for two mutations in trans configuration, a p.I382 M missense mutation and a p.V903GfsX3 frameshift deletion in both affected siblings. Electron microscopy of a skeletal muscle biopsy of the older sibling revealed dense osmiophilic bodies within the mitochondria. Mitochondrial DNA (mtDNA) content was within normal limits, and electron transport chain analysis showed no deficiencies of the mitochondrial respiratory chain enzymes. Multiple mtDNA deletions were not found. CONCLUSION: Compound heterozygosity of pathogenic OPA1 mutations may cause severe neuromuscular phenotypes in addition to early-onset optic atrophy. While a role for OPA1 in mtDNA maintenance has been discussed, compound biallelic pathogenic OPA1 mutations in our patients did not result in altered mtDNA copy number, mtDNA deletions, or deficiencies of the electron transport chain, despite the severe clinical phenotype. PMID- 21636303 TI - A ubiquitin ligase-associated chaperone holdase maintains polypeptides in soluble states for proteasome degradation. AB - Endoplasmic reticulum-associated degradation (ERAD) employs membrane-bound ubiquitin ligases and the translocation-driving ATPase p97 to retrotranslocate misfolded proteins for proteasomal degradation. How retrotranslocated polypeptides bearing exposed hydrophobic motifs or transmembrane domains (TMDs) avoid aggregation before reaching the proteasome is unclear. Here we identify a ubiquitin ligase-associated multiprotein complex comprising Bag6, Ubl4A, and Trc35, which chaperones retrotranslocated polypeptides en route to the proteasome to improve ERAD efficiency. In vitro, Bag6, the central component of the complex, contains a chaperone-like activity capable of maintaining an aggregation-prone substrate in an unfolded yet soluble state. The physiological importance of this holdase activity is underscored by observations that ERAD substrates accumulate in detergent-insoluble aggregates in cells depleted of Bag6, or of Trc35, a cofactor that keeps Bag6 outside the nucleus for engagement in ERAD. Our results reveal a ubiquitin ligase-associated holdase that maintains polypeptide solubility to enhance protein quality control in mammalian cells. PMID- 21636304 TI - [Emotional climate and internal communication in a clinical management unit compared with two traditional hospital services]. AB - BACKGROUND AND OBJECTIVE: The aim of this study is to compare the emotional climate, quality of communication and performance indicators in a clinical management unit and two traditional hospital services. MATERIAL AND METHODS: Quantitative study. INSTRUMENT: questionnaire of 94 questions. PARTICIPANTS: 83 health professionals (63 responders) from the clinical management unit of breast pathology and the hospital services of medical oncology and radiation oncology. ANALYSIS: descriptive statistics, comparison of means, correlation and linear regression models. RESULTS: The clinical management unit reaches higher values compared with the hospital services about: performance indicators, emotional climate, internal communication and evaluation of the leadership. An important gap between existing and desired sources, channels, media and subjects of communication appear, in both clinical management unit and traditional services. CONCLUSIONS: The clinical management organization promotes better internal communication and interpersonal relations, leading to improved performance indicators. PMID- 21636305 TI - Evaluation of enzyme immunoassay techniques for diagnosis of the most common intestinal protozoa in fecal samples. AB - OBJECTIVE: This study was designed to evaluate the antigen capture enzyme immunoassays (EIAs) Triage parasite panel and TechLab Entamoeba histolytica II in detecting Giardia intestinalis, Cryptosporidium sp, and Entamoeba histolytica in fecal samples in comparison to microscopy, and in differentiating Entamoeba histolytica from Entamoeba dispar. METHODS: The Triage EIA was evaluated using 100 stool specimens that were tested by standard ova and parasite examination, including staining with both trichrome and modified acid-fast stains. Differentiation between E. histolytica and E. dispar was performed using TechLab. RESULTS: Microscopic examination revealed that 19% of the samples were positive for Giardia, 4% for Cryptosporidium, and 1% for E. histolytica/E. dispar, and other parasites were found in 5%. By Triage, 23% of the samples were infected with Giardia, 5% with Cryptosporidium, and 2% with E. histolytica/E. dispar. Triage showed a sensitivity of 100% and specificity of 91.5%. The TechLab assay was negative for both samples diagnosed as E. histolytica/E. dispar by Triage, which suggested that they were E. dispar. Both tests showed no cross-reactivity with other intestinal protozoa. CONCLUSION: These results indicate that antigen detection by EIA has the potential to become a valuable tool, capable of making stool diagnostics more effective. PMID- 21636306 TI - Pigment epithelium-derived factor inhibits advanced glycation end-products induced cytotoxicity in retinal pericytes. AB - AIM: This study investigated the effects of pigment epithelium-derived factor (PEDF) on advanced glycation end-product (AGE)-induced cytotoxicity in porcine retinal pericytes and the signalling mechanism involved. METHODS: Retinal pericytes were isolated from porcine eyes and characterized by immunocytochemistry. The effect of AGEs and PEDF on cell proliferation was determined by bromodeoxyuridine (BrdU) assay. Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity was analyzed by luminescence assay. Reactive oxygen species (ROS), nitric oxide (NO), superoxide dismutase (SOD) and glutathione peroxidase (GSH) were determined by biochemical assays. Induction of apoptosis was determined by caspase-3 colorimetric assay and DNA fragmentation analysis. Src activity was assessed by transient transfection analysis, and the status of Src phosphorylation at Y419 was analyzed by a competitive ELISA method. RESULTS: AGEs significantly increased intracellular ROS generation in pericytes via NADPH oxidase and induced cell death via caspase-3 enzyme activation, whereas PEDF increased cell proliferation in a dose-dependent manner. In addition, PEDF inhibited AGE-induced ROS generation by increasing levels of SOD and GSH, and also blocked the activation of caspase-3. Furthermore, PEDF induced cell survival via the Src pathway by Src phosphorylation at Y419, as evidenced by a pharmacological inhibitor and Src mutants. CONCLUSION: These results suggest that PEDF abrogates AGE-induced oxidative stress and apoptosis in retinal pericytes via the Src pathway, thereby suggesting that PEDF is an effective therapeutic agent for the treatment of loss of pericytes in early diabetic retinopathy. PMID- 21636307 TI - Evaluation the pozzolanic reactivity of sonochemically fabricated nano natural pozzolan. AB - Natural pozzolans are appropriate supplementary cementitious materials in cement and concrete industry. A simple sonochemical method was developed to synthesize nanostructures of natural pozzolan. Chemical composition, crystallinity, morphology and reactivity of the natural pozzolan samples were compared before and after the sonochemical process, by using powder X-ray diffraction (XRD), Scanning Electron Microscopy (SEM), Thermal Gravimetry and Differential Thermal Analysis (TG/DTA). Compressive strength tests were performed to evaluate the properties of blended cements incorporating nano natural pozzolan. Under optimized conditions, the nano natural pozzolans showed a superior reactivity as compared with the bulk natural pozzolan. Also higher compressive strength was obtained for the cement specimen incorporating nano natural pozzolan. PMID- 21636308 TI - Activity classification using a single chest mounted tri-axial accelerometer. AB - Accelerometer-based activity monitoring sensors have become the most suitable means for objective assessment of mobility trends within patient study groups. The use of minimal, low power, IC (integrated circuit) components within these sensors enable continuous (long-term) monitoring which provides more accurate mobility trends (over days or weeks), reduced cost, longer battery life, reduced size and weight of sensor. Using scripted activities of daily living (ADL) such as sitting, standing, walking, and numerous postural transitions performed under supervised conditions by young and elderly subjects, the ability to discriminate these ADL were investigated using a single tri-axial accelerometer, mounted on the trunk. Data analysis was performed using Matlab(r) to determine the accelerations performed during eight different ADL. Transitions and transition types were detected using the scalar (dot) product technique and vertical velocity estimates on a single tri-axial accelerometer was compared to a proven discrete wavelet transform method that incorporated accelerometers and gyroscopes. Activities and postural transitions were accurately detected by this simplified low-power kinematic sensor and activity detection algorithm with a sensitivity and specificity of 86-92% for young healthy subjects in a controlled setting and 83-89% for elderly healthy subjects in a home environment. PMID- 21636309 TI - Development and validity of an instrumented handbike: initial results of propulsion kinetics. AB - OBJECTIVE: To develop an instrumented handbike system to measure the forces applied to the handgrip during handbiking. DESIGN: A 6 degrees of freedom force sensor was built into the handgrip of an attach-unit handbike, together with two optical encoders to measure the orientation of the handgrip and crank in space. Linearity, precision, and percent error were determined for static and dynamic tests. RESULTS: High linearity was demonstrated for both the static and the dynamic condition (r=1.01). Precision was high under the static condition (standard deviation of 0.2N), however the precision decreased with higher loads during the dynamic condition. Percent error values were between 0.3 and 5.1%. CONCLUSION: This is the first instrumented handbike system that can register 3 dimensional forces. It can be concluded that the instrumented handbike system allows for an accurate force analysis based on forces registered at the handle bars. PMID- 21636310 TI - Significance of cerebellar Purkinje cell loss to pathogenesis of essential tremor. AB - Some recent studies indicate that most essential tremor (ET) cases have cerebellar pathology characterized by Purkinje cell (PC) loss and its sequelae. OBJECTIVE: To assess the role of PC degeneration in ET symptomatology. METHODS: We studied seven ET, six tremor dominant Parkinson's disease controls and two normal control brains. Cerebellar PC counts were done in all cases by a neuropathologist using three different methods to identify PC. RESULTS: There were individual differences in PC counts in all subgroups. There was no difference between ET and controls of the same age. The PC count in ET was not related to: onset site of ET, severity of symptoms, or any other clinical features that we studied but there was a trend to a reduced number of PC with age in all groups. CONCLUSION: There is no evidence in our study or in the literature to date indicating that PC loss is the pathological basis of ET. Further studies are needed to determine the pathophysiology of ET. PMID- 21636311 TI - Reconstruction of scattered data in fetal diffusion MRI. AB - In this paper we present a method for reconstructing diffusion-weighted MRI data on regular grids from scattered data. The proposed method has the advantage that no specific diffusion model needs to be assumed. Previous work assume the tensor model, but this is not suitable under certain conditions like intravoxel orientational heterogeneity (IVOH). Data reconstruction is particularly important when studying the fetal brain in utero, since registration methods applied for movement and distortion correction produce scattered data in spatial and diffusion domains. We propose the use of a groupwise registration method, and a dual spatio-angular interpolation by using radial basis functions (RBF). Leave one-out experiments performed on adult data showed a high accuracy of the method. The application to fetal data showed an improvement in the quality of the sequences according to objective criteria based on fractional anisotropy (FA) maps, and differences in the tractography results. PMID- 21636312 TI - The effect of hydrogel and silicone hydrogel contact lenses on the measurement of intraocular pressure with rebound tonometry. AB - PURPOSE: To assess the accuracy of intraocular pressure (IOP) measurements using rebound tonometry over disposable hydrogel (etafilcon A) and silicone hydrogel (senofilcon A) contact lenses (CLs) of different powers. METHODS: The experimental group comprised 36 subjects (19 male, 17 female). IOP measurements were undertaken on the subject's right eyes in random order using a rebound tonometer (ICare). The CLs had powers of +2.00D, -2.00D and -6.00D. Six measurements were taken over each contact lens and also before and after the CLs had been worn. RESULTS: A good correlation was found between IOP measurements with and without CLs (all r>=0.80; p<0.05). Bland Altman plots did not show any significant trend in the difference in IOP readings with and without CLs as a function of IOP value. A two-way ANOVA revealed a significant effect of material and power (p<0.01) but no interaction. All the comparisons between the measurements without CLs and with hydrogel CLs were significant (p<0.01). The comparisons with silicone hydrogel CLs were not significant. CONCLUSIONS: Rebound tonometry can be reliably performed over silicone hydrogel CLs. With hydrogel CLs, the measurements were lower than those without CLs. However, despite the fact that these differences were statistically significant, their clinical significance was minimal. PMID- 21636313 TI - The involvement of CHD5 hypermethylation in laryngeal squamous cell carcinoma. AB - Chromodomain helicase DNA-binding protein 5 (CHD5) has been found to be a candidate tumor suppressor gene (TSG) in malignant neural tumors. In mice heterozygous for chd5 deficiency, the first tumor observed was pathological squamous cell carcinoma. More than 95% of primary laryngeal cancer is squamous cell carcinoma. Thus, we explored the expression of CHD5 in 65 patients with laryngeal squamous cell carcinoma (LSCC) using real-time PCR, immunohistochemistry and Western blotting. DNA methylation was detected using bisulfate-specific sequencing. The potential function of CHD5 was determined using MTT, apoptosis and transwell migration assays in CHD5-transfected Hep-2 cells. Our results revealed that the mRNA and protein expression levels of CHD5 in LSCC tissues were significantly lower than those in clear surgical margin tissues (p<0.05), and there is a significant correlation between the mRNA and protein expression levels of CHD5 (p<0.01). In addition, there were significant differences in CHD5 mRNA and protein levels with respect to the patient's clinical stage (p<0.05). Aberrant methylation of the CHD5 promoter was frequently found in the Hep-2 cell line and LSCC tumor tissues, especially tumor tissues from advanced TNM (p<0.05) or older patients (p<0.05). Finally, ectopic expression of CHD5 in laryngeal cancer cells led to significant inhibition of growth and invasiveness. Our data suggest that CHD5 is a tumor suppressor gene that is epigenetically downregulated in LSCC. PMID- 21636314 TI - Endogenous peptide elicitors in higher plants. AB - Plant defense responses against invading organisms are initiated through the perception of molecules associated with attacking microbes and herbivores by pattern recognition receptors. In addition to elicitor molecules derived from attacking organisms, plants recognize host-derived molecules. These endogenous elicitors induce and amplify the defense responses against invading organisms both locally and systemically. Several classes of plant-derived molecules elicit defense, including cell wall fragments and peptides. Endogenous peptide elicitors have been discovered in species across the plant kingdom, and their role regulating immunity to both herbivores and pathogens is becoming increasingly appreciated. In this review, we will focus on the five known endogenous peptide elicitor families, summarize their properties, and discuss research goals to further understanding of plant innate immunity. PMID- 21636315 TI - Course of poly(4-aminodiphenylamine)/Ag nanocomposite formation through UV-vis spectroscopy. AB - Kinetics of chemical oxidative polymerization of 4-aminodiphenylamine (4ADPA) was followed in aqueous 1 M p-toluene sulfonic acid (p-TSA) using silver nitrate (AgNO3) as an oxidant by UV-vis spectroscopy. The medium was found to be clear and homogeneous during the course of polymerization. The absorbances corresponding to the intermediate and the polymer were followed for different concentrations of 4ADPA and AgNO3 and at different reaction time. The appearance of a band around 450 nm during the initial stages of polymerization corresponds to the plasmon resonance formed by the reduction of Ag+ ions. Rate of poly(4 aminodiphenylamine)/Ag nanocomposite (RP4ADPA/AgNC) was determined for various reaction conditions. R(P4ADP/AgNC) showed second order power dependence on 4ADPA and first order dependence on AgNO3. The observed order dependences of 4ADPA and AgNO3 on the formation of P4ADPA/AgNC were used to deduce a rate equation for the reaction. Rate constant for the reaction was determined through different approaches. The good agreement between the rate constants obtained through different approaches justifies the selection of rate equation. PMID- 21636316 TI - Crystal growth and characterization of gamma-glycine grown from potassium fluoride for photonic applications. AB - Single crystals of gamma-glycine, an organic nonlinear optical material have been synthesized in the presence of potassium fluoride (KF) by slow evaporation technique at ambient temperature. The size of the grown crystal is up to the dimension of 12 mm*10 mm*8 mm. The gamma-phase was confirmed by single crystal X ray diffraction, powder XRD and the FTIR analysis. Optical absorption spectrum reveals that the grown crystal has good optical transparency in the entire visible region with an energy band gap of 5.09 eV, which is an essential requirement for a nonlinear optical crystal. Thermal stability of the grown gamma glycine crystal was determined using the thermo gravimetric and differential thermal analyses. The NLO activity of gamma-glycine was confirmed by the Kurtz powder technique using Nd:YAG laser and the grown crystal exhibits high relative conversion efficiency when compared to potassium dihydrogen phosphate (KDP). PMID- 21636317 TI - Diagnosis of dengue fever in North West Italy in travelers from endemic areas: a retrospective study. AB - BACKGROUND: Domestic outbreaks of Dengue (DENV) fever from imported cases have to be considered a possible risk in non-endemic countries where Dengue vectors are present, such as in Italy. OBJECTIVE: To review imported acute/recent DENV infections in a one-year survey in a North West Italy region where the presence of Aedes albopictus is documented. STUDY DESIGN: We retrospectively reviewed laboratory and clinical records of Italian febrile travelers from Dengue endemic areas referring to the local reference Centre for Infectious Disease, covering a population of about 4 million people. RESULTS: Acute/recent DENV infection was identified in 15 out of 91 travelers from endemic areas (16.5%) including 12 primary and 3 secondary infections; in 6 patients the virus was detectable in blood according to molecular real-time Polymerase Chain Reaction-based assays: in 9 patients the diagnosis of DENV infection was accomplished by the combination of specific IgM reactivity, high IgG titers, IgG seroconversion from negative to positive and increasing (four-fold) IgG titers in paired serum samples. Two cases of DENV infections were imported from South Egypt in patients travelling together, confirming the importance of returning travelers as sentinels of a rapidly changing epidemiology in specific geographic areas. CONCLUSIONS: Our findings outline the high rate of imported Dengue infection in North West Italy and emphasize the need for a continued Dengue surveillance in non-endemic countries as well as a careful evaluation and follow-up of febrile patients returning from Dengue endemic countries. PMID- 21636318 TI - Human parainfluenza virus type 4 infection in Chinese children with lower respiratory tract infections: a comparison study. AB - BACKGROUND: Human parainfluenza viruses (HPIVs) are a leading cause of acute respiratory tract infections (ARTIs). Although HPIV-4 has been associated with mild ARTIs for years, recent investigations have also associated HPIV-4 infection with severe respiratory syndromes and with outbreaks of ARTIs in children. OBJECTIVES: To characterize the role of HPIV-4 and its clinical features in children with acute lower respiratory tract infections (ALRTIs) in Beijing, China. STUDY DESIGN: Nasopharyngeal aspirates were collected from 2009 hospitalized children with ALRTIs between March 2007 and April 2010. RT-PCR and PCR analyses were used to identify HPIV types and other known respiratory viruses. RESULTS: HPIVs were detected in 246 (12.2%) patients, of whom 25 (10.2%) were positive for HPIV-4, 11 (4.5%) for HPIV-2, 51 (20.7%) for HPIV-1, 151 (61.4%) for HPIV-3, and 8 (3.3%) were co-detected with different types of HPIVs. Like HPIV-3, HPIV-4 was detected in spring, summer, and late fall over the study period. Seasonal incidence varied for HPIV-1 and -2. The median patient age was 20 months for HPIV-4 infections and 7-11 months for HPIV-1, -2, and -3 infections, but the clinical manifestations did not differ significantly between HPIV-1, -2, -3, and -4 infections. Moreover, co-detection of HPIV-4 (44%) with other respiratory viruses was lower than that of HPIV-1 (62.7%), HPIV-2 (63.6%), and HPIV-3 (72.7%). CONCLUSIONS: HPIV-4 plays an important role in Chinese paediatric ALRTIs. The epidemiological and clinical characteristics reported here improve our understanding of the pathogenesis associated with HPIV-4. PMID- 21636319 TI - The utility of EMG interference pattern analysis in botulinum toxin treatment of torticollis: a randomised, controlled and blinded study. AB - OBJECTIVE: The significance of electromyography (EMG) guidance in botulinum toxin (BT) treatment has been much debated. The aim of this study was to evaluate if EMG guidance in the treatment of torticollis in BT-naive patients had a better outcome than treatment after clinical evaluation alone. METHODS: Twenty-six patients with torticollis were included and treated for 1 year in this prospective, blinded study. Quantitative EMG was performed simultaneously in the four most frequently affected muscles: the sternocleidomastoid muscles and the posterior neck muscles on both sides. EMGs were analysed for turns per second. Clinical ratings were performed by an experienced neurologist (A). Injections were given by another neurologist (B), who was blinded to the ratings. In group 1, the results of the EMG were available to the treating neurologist B, whereas in group 2, neurologist B was blinded. In group 1, treatment with BT was given when turns per second were higher than 100. RESULTS: In patients treated guided by EMG, clinical outcome, evaluated by objective ratings, was better than in patients treated based on clinical judgement alone (p = 0.05). In group 2, 105 muscles were treated with BT. Of these, 37 did not show dystonic EMG activity. CONCLUSIONS: Treatment with BT guided by EMG results in better clinical outcome than treatment without EMG and reduces the amount of BT used. SIGNIFICANCE: EMG guidance by interference pattern analysis may optimise BT treatment in torticollis by a more precise injection and may reduce side effects and the risk of development of antibodies to BT. PMID- 21636320 TI - Flow cytometric analysis of lectin-oligosaccharide interactions by using fluorescent oligosaccharide probes. AB - Tetraphenylethylene (TPE) derivatives have strong fluorescence in aggregated state. We report here an oligosaccharide binding assay system using tetraphenylethylene derivatives bearing oligosaccharides with aggregation-induced emission (AIE) characteristics. A tetraphenylethylene derivative bearing 6' sialyllactose (6'SL) bound to microbeads coated with SSA lectin more effectively than RCA120 lectin. Microbeads that bound to fluorescent oligosaccharide probes could be detected by flow cytometric analysis. Tetraphenylethylene derivatives bearing oligosaccharides are useful for flow cytometric analysis of lectin oligosaccharide interactions. PMID- 21636321 TI - A randomized controlled trial for the effect of passive stretching on measures of hamstring extensibility, passive stiffness, strength, and stretch tolerance. AB - To measure hamstring extensibility, stiffness, stretch tolerance, and strength following a 4-week passive stretching program. Randomized controlled trial. Twenty-two healthy participants were randomly assigned to either a 4-week stretching program consisting of 4 hamstring and hip stretches performed 5 times per week, or a non-stretching control group. Hamstring extensibility and stiffness were measured before and after training using the instrumented straight leg raise test (iSLR). Stretch tolerance was measured as the pain intensity (visual analog scale; VAS) elicited during the maximal stretch. Hamstring strength was measured using isokinetic dynamometry at 30 and 120 degrees s(-1). Hamstring extensibility increased by 20.9% in the intervention group following 4 weeks of training (p<0.001; d=0.86). Passive stiffness was reduced by 31% in the intervention group (p<0.05; d=-0.89). Stretch tolerance VAS scores were not different between groups at either time point, and no changes were observed following training. There were no changes in hamstring concentric strength measured at 30 and 120 degrees s(-1). Passive stretching increases hamstring extensibility and decreases passive stiffness, with no change in stretch tolerance defined by pain intensity during the stretch. Compared to previous research, the volume of stretching was higher in this study. The volume of prescribed stretching is important for eliciting the strong clinical effect observed in this study. PMID- 21636322 TI - Knee and hip sagittal and transverse plane changes after two fatigue protocols. AB - Fatigue has been shown to alter the biomechanics of lower extremity during landing tasks. To date, no study has examined the effects of two types of fatigue on kinetics and kinematics. OBJECTIVES: This study was conducted to assess biomechanical differences between two fatigue protocols [Slow Linear Oxidative Fatigue Protocol (SLO-FP) and Functional Agility Short-Term Fatigue Protocol (FAST-FP)]. DESIGN: Single-group repeated measures design. METHODS: Fifteen female collegiate soccer players had to perform five successful trials of unanticipated sidestep cutting (SS) pre- and post-fatigue protocols. The SLO-FP consisted of an initial VO(2peak) test followed by 5-min rest, and a 30-min interval run. The FAST-FP consisted of 4 sets of a functional circuit. Biomechanical measures of the hip and knee were obtained at different instants while performing SS pre- and post-fatigue. Repeated 2 * 2 ANOVAs were conducted to examine task and fatigue differences. Alpha level set a priori at 0.05. RESULTS: During the FAST-FP, participants had increased knee internal rotation at initial contact (IC) (12.5 +/- 5.9 degrees ) when compared to the SLO-FP (7.9 +/- 5.4 degrees , p<0.001). For hip flexion at IC, pre-fatigue had increased angles (36.4 +/- 8.4 degrees ) compared to post-fatigue (30.4 +/- 9.3 degrees , p=0.003), also greater knee flexion during pre-fatigue (25.6 +/- 6.8 degrees ) than post-fatigue (22.4 +/- 8.4 degrees , p=0.022). CONCLUSION: The results of this study showed that hip and knee mechanics were substantially altered during both fatigue conditions. PMID- 21636323 TI - Regulatory T cell plasticity: beyond the controversies. AB - Forkhead box P3 (Foxp3)(+) regulatory T (Treg) cells represent a distinct cell lineage that is committed to suppressive functions, whose stable differentiation state ensures the robustness of self-tolerance and immune homeostasis in a changing environment. Recent studies have challenged this notion and suggest that Treg cells retain developmental plasticity to be reprogrammed to Foxp3(-) helper T cells in response to extrinsic perturbations such as inflammation and lymphopenia. This issue of Treg cell plasticity, however, remains controversial because other recent reports argue against the plasticity phenomena. Here, I propose that the controversies can be resolved by considering the heterogeneity model of plasticity, which hypothesizes that the observed plasticity does not reflect lineage reprogramming of Treg cells but rather a minor population of uncommitted Foxp3(+) T cells. PMID- 21636324 TI - Mycobacterium tuberculosis infection induces hypoxic lung lesions in the rat. AB - Hypoxia is believed to influence the metabolic state of Mycobacterium tuberculosis and cause phenotypic drug resistance. Using pimonidazole adduct staining, we show that lung lesions of infected rats contain regions of low oxygen tension. Our results support the use of the rat model for evaluating anaerobic drug activity in vivo. PMID- 21636325 TI - Epidemiology, antibiotic resistance trends and the cost of enteric fever in East London, 2005-2010. AB - INTRODUCTION: Enteric fever seen in the UK has usually been acquired abroad. The cost to the NHS of treating enteric fever cases is not known. Data on the epidemiology of enteric fever, inpatient treatment costs and the public health management is needed to make decisions regarding the cost benefit considerations of introducing targeted prevention strategies. METHODS: A retrospective study of laboratory confirmed enteric fever cases was conducted to estimate the cost of inpatient treatment and to determine antimicrobial resistance patterns at two hospitals in East London between January 2005 and the end of August 2010. RESULTS: 138 cases of enteric fever were identified during the study period (90 S.ser.Typhi and 48 S. ser. Paratyphi). 92% had a recent history of foreign travel, 57% had travelled to visit friends and relatives (VFRs), 26% sought pre travel health advice and 26% of patients had received typhoid vaccination. The inpatient treatment cost of 138 cases to the NHS was L272,747. The proportion of isolates with high level ciprofloxacin resistance (MICs>1 mg/L) has increased from 10% in 2006 to 30% in 2010. Our data also shows the emergence of isolates with high azithromycin MICs (>32 mg/L); 60% (six out of ten) isolates tested in July-August 2010. CONCLUSIONS: There is a significant direct cost of treating enteric fever cases on the NHS. Cost reduction measures are confined due to the lack of effective oral antibiotics following the emergence of high level resistance to ciprofloxacin and azithromycin. Outpatient parenteral antibiotic therapy service and improved preventative public health measures aimed at VFR travellers in particular may be helpful in reducing costs. PMID- 21636326 TI - Parental information, motivation, and behavioral skills correlate with child sweetened beverage consumption. AB - OBJECTIVE: To evaluate fit of the Information-Motivation-Behavioral Skills (IMB) model applied to sweetened beverage (SB) consumption in children. DESIGN: Cross sectional. Parents completed a home beverage inventory and IMB survey regarding SB consumption. SETTING: Health fairs, Special Supplemental Nutrition Program for Women, Infants, and Children clinics. PARTICIPANTS: Convenience sample of 198 parents of low socioeconomic status. MAIN OUTCOME MEASURES: Independent variables included scores from 3 indices calculated from the IMB survey, information, motivation, and behavioral skills. The dependent variable was average child daily caloric consumption from SB consumption calculated from the home beverage inventory. ANALYSIS: Structural equation modeling. RESULTS: Parental information had direct and indirect negative relationships with SB consumption. Parental motivation was only indirectly associated with SB consumption mediated through behavioral skills. Parental behavioral skills had a negative correlation with SB consumption. CONCLUSIONS AND IMPLICATIONS: In applying the IMB model to SB consumption, the authors found preliminary support for relationships between parental information, motivation, behavioral skills, and child SB consumption. Application of this model shows promise in identifying the complex interactions between factors influencing consumption behaviors. As nutrition educators frequently desire to alter behaviors, use of the IMB model may help with both targeting and assessment efforts. PMID- 21636327 TI - Lack of increased prevalence of immunoregulatory disorders in hereditary angioedema due to C1-inhibitor deficiency. AB - Hereditary angioedema due to deficiency of C1-INH (HAE-C1-INH) is associated with enhanced consumption of the early complement components, which may predispose for autoimmune disease. We assessed the prevalence of such disorders among HAE- C1 INH patients and their impact on the natural course of HAE-C1-INH. Clinical data and immunoserological parameters of 130 HAE-C1-INH and 174 non-C1-INH-deficient patients with angioedema were analyzed. In our study, the incidence of immunoregulatory disorders was 11.5% in the population of HAE-C1-INH patients and 5.2% among non-C1-INH-deficient controls with angioedema. Immunoserology screening revealed a greater prevalence of anticardiolipin IgM (p=0.0118) among HAE-C1-INH patients, than in those with non-C1-INH-deficient angioedema. We did not find higher prevalence of immunoregulatory disorders among our HAE-C1-INH patients. However, in patients with confirmed immunoregulatory disorders, the latter influenced both the severity of HAE-C1-INH and the effectiveness of its long-term management. Appropriate management of the immunoregulatory disease thus identified improves the symptoms of HAE-C1-INH. PMID- 21636328 TI - Histology of cervical intraepithelial neoplasia and the role of biomarkers. AB - Accurate histological grading of cervical intraepithelial neoplasia (CIN) lesions is important for clinical management of patients, because CIN1 and CIN2 and 3 lesions are treated differently. In general, there tends to be poor inter and intra-observer reproducibility of CIN grade evaluation among pathologists. In particular, the differential diagnosis between immature squamous metaplasia and CIN1 and 2, or between low-grade (CIN1) and high-grade (CIN2 and 3) lesions, tend to be difficult. These difficulties mean that patients tend to be over-treated for CIN lesions, which will naturally regress. Collectively, this highlights the need for alternative approaches and specific biomarkers to aid objective CIN lesion grading, and to identify true high-grade cervical disease. In this review we focus on the aetiology, pathobiology, the natural history of CIN, current issues with diagnosis and classification of CIN and the diagnostic and prognostic utility of specific biomarkers in identifying true cancerous precursor lesions. PMID- 21636329 TI - Cardiovascular and neuroendocrine features of Panayiotopoulos syndrome. PMID- 21636330 TI - Number, size, conduction, and vasoconstrictor ability of unmyelinated fibers of the ovarian nerve in adult and aged rats. AB - The effect of aging on the number, size, conduction velocity, and vasoconstrictive function of unmyelinated fibers in ovarian nerve accompanying the ovarian artery was studied in adult (4-7mo) and aged (28-31mo) rats. Morphological observation by electron microscopy showed that the ovarian nerve contains mainly unmyelinated fibers with only a small percentage (less than 4%) of myelinated fibers in either age group. The number of unmyelinated fibers tended to decrease in aged rats (717+/-59) compared to adult rats (801+/-48), especially in fibers of smaller diameter, although this difference was not statistically significant. The maximum conduction velocity of unmyelinated fibers within the ovarian nerve was similar when compared between adult (1.05+/-0.04m/s) and aged (1.02+/-0.05m/s) rats. Under anesthesia, electrical stimulation of the distal portion of a severed ovarian nerve reduced ovarian blood flow, as measured by laser Doppler flowmetry, when the stimulus intensity was above the threshold for unmyelinated C fibers. Stimulation of the ovarian nerve with supra-maximum intensity (10V) at 2-20Hz frequencies produced frequency-dependent reductions in ovarian blood flow in both adult and aged rats. There were no significant differences in magnitude of the reduction in ovarian blood flow with comparable frequencies of electrical stimulation of the ovarian nerve between adult and aged rats. Collectively, these data indicate that unmyelinated C fibers in ovarian nerve are maintained in number, size, conduction ability, and vasoconstrictor function in aged rats. PMID- 21636331 TI - Staphylococcus aureus with decreased susceptibility to glycopeptides in cystic fibrosis patients. AB - We report the isolation of Staphylococcus aureus with decreased susceptibility to glycopeptides in five CF patients and review the clinical and microbiological features of these cases. Three patients presented with pulmonary exacerbation that may be attributed to these strains and two of them were successfully treated using linezolid therapy. Glycopeptide-intermediate S. aureus (GISA) strains isolated in two patients were susceptible to methicillin, while the three other patients harbored methicillin-resistant GISA. Rarely reported in CF, GISA may remain underestimated due to the difficulty of detection, and both clinicians and microbiologists should be aware of the GISA emergence in CF patients' population. PMID- 21636332 TI - UV B-irradiation enhances the racemization and isomerizaiton of aspartyl residues and production of Nepsilon-carboxymethyl lysine (CML) in keratin of skin. AB - UV-B irradiation is one of the risk factors in age-related diseases. We have reported that biologically uncommon D-beta-Asp residues accumulate in proteins from sun-exposed elderly human skin. A previous study also reported that carboxymethyl lysine (CML; one of the advanced glycation end products (AGEs)) which is produced by the oxidation of glucose and peroxidation of lipid, also increases upon UV B irradiation. The formation of D-beta-Asp and CML were reported as the alteration of proteins in UV B irradiated skin, independently. In this study, in order to clarify the relationship between the formation of D-beta Asp and CML, immunohistochemical analysis using anti-D-beta-Asp containing peptide antibodies and anti-CML antibodies was performed in UV B irradiated mice. Immunohistochemical analyses clearly indicated that an anti-D-beta-Asp containing peptide antibody and anti-CML antibody reacted at a common area in UV B irradiated skin. Western blot analyses of the proteins isolated from UV B irradiated skin demonstrated that proteins of 50-70 kDa were immunoreactive towards antibodies for both D-beta-Asp containing peptide and CML. These proteins were identified by proteomic analysis as members of the keratin families including keratin-1, keratin-6B, keratin-10, and keratin-14. PMID- 21636333 TI - Signification and significance: Music, brain, and culture: Comment on "Towards a neural basis of processing musical semantics" by S. Koelsch. PMID- 21636334 TI - Myth: cerebral palsy cannot be predicted by neonatal brain imaging. AB - There is controversy in the literature about the value of brain imaging in neonates regarding the prediction of cerebral palsy (CP). The aim of this review was to unravel the myth that CP cannot be predicted by neuroimaging in neonates. Major intracranial lesions in the preterm infant should be recognized with sequential cranial ultrasound and will predict those with non-ambulatory CP. Magnetic resonance imaging (MRI) at term-equivalent age will refine the prediction by assessment of myelination of the posterior limb of the internal capsule. Prediction of motor outcome in preterm infants with subtle white matter injury remains difficult, even with conventional MRI. MRI is a better tool to predict outcome in the term infant with hypoxic-ischaemic encephalopathy or neonatal stroke. The use of diffusion-weighted imaging as an additional sequence adds to the predictive value for motor outcome. Sequential and dedicated neuroimaging should enable us to predict motor outcome in high risk newborns infants. PMID- 21636335 TI - Myth: neonatology is evidence-based. AB - The practice of evidence-based medicine involves the judicious use of current best evidence in the care of individual patients. Decisions about diagnosis, prognosis and treating patients require knowledge of the probability and value of outcomes. Decision analysis illustrates how probabilities and values help define one another, and each are important. Whereas initial probability estimates can be obtained by 'searching for the best evidence', values belong to individuals. Obtaining values from patients or parents is sometimes difficult and requires a respectful, thoughtful, systematic approach, but only after doing this is neonatal care evidence-based. PMID- 21636336 TI - Long-term results after 40 years experience with treatment of rare facial clefts: Part 1--Oblique and paramedian clefts. AB - BACKGROUND: Oblique and paramedian rare facial clefts impose a major reconstructive challenge and long-term assessments of the outcomes remain scarce. This study provides new details regarding surgical techniques and timing, influence of growth, and difficulties of this pathology on the long-term; a guideline for surgical treatment is given. METHODS: Twenty-nine adults with an oblique or paramedian facial cleft and surgically treated in the authors' unit between 1969 and 2009, were included. The long-term evaluation was based on series of photographs, 3D-CT's, X-rays, operation data, and was specified per facial area. RESULTS: The mean number of performed operations per patient was 10.6 (range: 1-26). Vertical dystopia is not caused by previous surgery, but by growth deficiencies of the maxilla. In all patients with vertical dystopia, its presence and severity were clear at the age of five, and it should ideally be treated shortly after that age. In mild cases grafting seems sufficient, but in more severe cases orbital translocation is necessary. Costochondral grafts showed the best long-term results in both orbital and nasal reconstructions. Major nose reconstruction is best delayed until adolescence. For an optimal final result in selected cases, correction of midface hypoplasia at adolescence is necessary. CONCLUSION: The three-dimensional underdevelopment of the midface region plays a central role in the deformities of most patients, but is complex and difficult to correct. The provided guideline should help to minimize the number of operations and ameliorate long-term results. PMID- 21636337 TI - The use of a template to accurately position the inframammary fold in breast reconstruction. AB - Reconstruction of the inframammary fold (IMF) during breast reconstruction is vital in achieving good aesthetic outcomes and avoiding further corrective surgery. The IMF is usually reconstructed by comparing with the other side and estimating its correct position. The IMF template can be used to accurately and consistently mark the position of the IMF relative to the contralateral side thus aiding accurate reconstruction without the need to estimate. PMID- 21636338 TI - Ultrastructural changes in cardiac myocytes from Boxer dogs with arrhythmogenic right ventricular cardiomyopathy. AB - OBJECTIVES: We sought to quantify the number and length of desmosomes, gap junctions, and adherens junctions in arrhythmogenic right ventricular cardiomyopathy (ARVC) and non-ARVC dogs, and to determine if ultrastructural changes existed. ANIMALS: Hearts from 8 Boxer dogs afflicted with histopathologically confirmed ARVC and 6 dogs without ARVC were studied. METHODS: Quantitative transmission electron microscopy (TEM) and Western blot semi quantification of alpha-actinin were used to study the intercalated disc and sarcomere of the right and left ventricles. RESULTS: When ARVC dogs were compared to non-ARVC dogs reductions in the number of desmosomes (P = 0.04), adherens junctions (P = 0.04) and gap junctions (P = 0.02) were found. The number of gap junctions (P = 0.04) and adherens junctions (P = 0.04) also were reduced in the left ventricle, while the number of desmosomes was not (P = 0.88). A decrease in the length of desmosomal complexes within LV samples (P = 0.04) was found. These findings suggested disruption of proteins providing attachment of the cytoskeleton to the intercalated disc. Immunoblotting did not demonstrate a quantitative reduction in the amount of alpha-actinin in ARVC afflicted samples. All Boxers with ARVC demonstrated the presence of electron dense material originating from the Z band and extending into the sarcomere, apparently at the expense of the cytoskeletal structure. CONCLUSIONS: These results emphasize the importance of structural integrity of the intercalated disc in the pathogenesis of ARVC. In addition, observed abnormalities in sarcomeric structure suggest a novel link between ARVC and the actin-myosin contractile apparatus. PMID- 21636339 TI - Congenital coronary-pulmonary arterial shunt in a German shepherd dog: diagnosis and surgical correction. AB - BACKGROUND: A 10 week-old intact female German shepherd dog was examined because of a heart murmur. METHODS: An echocardiogram revealed an extracardiac left-to right shunt. An angiogram identified shunting between the aorta and the pulmonary artery in an unusual location. RESULTS: Thoracotomy was performed to better identify and correct the lesion; a tubular shunt between the left coronary artery and the pulmonary artery was found and ligated in a closed-heart procedure. The murmur resolved immediately upon ligation and the dog's heart size normalized over a period of several months after surgery. CONCLUSION: In the dog, surgical correction of a coronary artery-pulmonary artery shunt can be performed without complications and can be associated with reversal of cardiac remodeling. PMID- 21636340 TI - [Primary hyperoxaluria]. AB - Primary hyperoxalurias are rare recessive inherited inborn errors of glyoxylate metabolism. They are responsible for progressive renal involvement, which further lead to systemic oxalate deposition, which can even occur in infants. Primary hyperoxaluria type 1 is the most common form in Europe and is due to alanine glyoxylate aminostransferase deficiency, a hepatic peroxisomal pyridoxin dependent enzyme. Therefore primary hyperoxaluria type 1 is responsible for hyperoxaluria leading to aggressive stone formation and nephrocalcinosis. As glomerular filtration rate decreases, systemic oxalate storage occurs throughout all the body, and mainly in the skeleton. The diagnosis is first based on urine oxalate measurement, then on genotyping, which may also allow prenatal diagnosis to be proposed. Conservative measures - including hydration, crystallization inhibitors and pyridoxine - are safe and may allow long lasting renal survival, provided it is given as soon as the diagnosis has been even suspected. No dialysis procedure can remove enough oxalate to compensate oxalate overproduction from the sick liver, therefore a combined liver and kidney transplantation should be planned before advanced renal disease has occurred, in order to limit/avoid systemic oxalate deposition. In the future, primary hyperoxaluria type 1 may benefit from hepatocyte transplantation, chaperone molecules, etc. PMID- 21636341 TI - A paternity case with mutations at three CODIS core STR loci. PMID- 21636342 TI - Association analysis of intractable epilepsy with C3435T and G2677T/A ABCB1 gene polymorphisms in Iranian patients. AB - OBJECTIVE: The results from studies investigating a possible association between ABCB1 polymorphism and drug-resistant epilepsy are so far inconsistent. Moreover, recent meta-analyses studies do not confirm any link between ABCB1 C3435T polymorphism and drug resistance. Yet, if patients with comparable clinical status (same type of epilepsy, antiepileptic drugs, epilepsy onset and gender) are evaluated, the link between ABCB1 polymorphisms and drug resistance may be unmasked. We studied the association between C3435T and G2677T/A ABCB1 gene polymorphisms and drug resistance in Iranian epilepsy patients. METHODS: Two hundred healthy subjects and 332 epilepsy patients (200 drug-responsive and 132 drug-resistant) were selected. Genotypes were determined by polymerase chain reaction followed by restriction fragment length polymorphism or the amplification refractory mutation system. RESULTS: The risk of drug resistance was higher in patients with a C/T genotype than in those with C/C or T/T genotypes at position 3435 in patients with cryptogenic epilepsy (p=0.01). A higher risk of drug resistance was observed in adult patients with a C/C genotype than in those with a T/T genotype at position 3435 (25.8% vs 15.8%, p=0.01). The risk of drug resistance was also higher in female patients with a C/C genotype than in those with a T/T genotype at position 3435 (26.8% vs 16.3%, p=0.04). No significant association was found between G2677T/A polymorphism and epilepsy drug resistance in the different subgroups of patients. CONCLUSION: Iranian adult female patients with a C/C genotype at position 3435 of the ABCB1 gene have a higher risk of resistance to antiepileptic drugs. Replication studies with large sample sizes are needed to confirm the results. PMID- 21636343 TI - [How to discuss death with children and families?]. AB - Taking care of a child or an adolescent in palliative stage is a testing work for health professionals in paediatric onco-hematology. The communication with the child at the end of life and his family is delicate but however essential to ensure quality cares while supporting trust. The continuation of a genuine relation and a regular information concerning the child's health help the parents to progressively understand the lack of possible cure and the future death of their child. This anticipation also allows to possibly discuss about conditions and place of death of the ill child or adolescent. The child or adolescent himself at the end of life can also feel the need for speaking about his situation. It is then for the adult, parent or health professional, to share with him his questionings and his possible worries in order to reduce his loneliness and his fears. To let be guided by the child allows to respect his progression and his wish to know more about it or not. Finally a support must also be offered for the siblings. PMID- 21636344 TI - Radiation-induced modifications of the tumor microenvironment promote metastasis. AB - Radiotherapy is successfully used to treat cancer. Emerging evidence, however, indicates that recurrences after radiotherapy are associated with increased local invasion, metastatic spreading and poor prognosis. Radiation-induced modifications of the tumor microenvironment have been proposed to contribute to increased aggressive tumor behavior, an effect also referred to as tumor bed effect, but the putative mechanisms involved have remained largely elusive. We have recently demonstrated that irradiation of the prospective tumor stroma impairs de novo angiogenesis through sustained inhibition of proliferation, migration and sprouting of endothelial cells. Experimental tumors growing within a pre-irradiated field have reduced tumor angiogenesis and tumor growth, increased hypoxia, necrosis, local invasion and distant metastasis. Mechanisms of progression involve adaptation of tumor cells to local hypoxic conditions as well as selection of cells with invasive and metastatic capacities. The matricellular protein CYR61 and integrin alphaVbeta5 emerged as molecules that cooperate to mediate lung metastasis. Cilengitide, a small molecular inhibitor of alphaV integrins prevented lung metastasis formation. These results represent a conceptual advance to the understanding of the tumor bed effect and indicate that alphaV integrin inhibition might be a potential therapeutic approach for preventing metastasis in patients at risk for post-radiation recurrences. PMID- 21636345 TI - [Medical treatments of endocrine-sensitive Her-2 negative breast cancers: a review]. AB - New molecular classification is one of the cornerstones of current and future progress in research and patient care for breast carcinoma. For the larger hormone-receptor positive and Her-2 negative subgroup, which concerns 75% of the patients, endocrine therapy and chemotherapy may be considered. Looking toward new-targeted therapies, this paper reviews the current use of these two treatment modalities in adjuvant, neoadjuvant and metastatic settings of this disease. PMID- 21636346 TI - [Pathogenesis and management of refractory malignant ascites]. AB - Malignant ascites are the cancer-associated accumulation of fluids in the peritoneal cavity. The neoplasms most frequently associated with ascites are ovarian, breast, colon, stomach and pancreas adenocarcinomas. Symptoms are abdominal distention, nausea, vomiting, anorexia, dyspnea and limbs oedemas. Several pathophysiological mechanisms might be implicated such as peritoneal carcinomatosis, lymphatic vessels' obstruction, portal hypertension or heart failure. Its diagnosis is most often performed in a context of already known neoplasia. Malignant ascites are associated with a pejorative evolution. Ascites which cannot be mobilized or show early recurrence and cannot be prevented by medical treatment are defined as refractory ascites. Therefore, management of refractory malignant ascites takes place in the context of palliative care and aims at improving the quality of life of these patients. This review lists the current data reported on the pathophysiology of malignant ascites and describes the present and future options for refractory malignant ascites management. PMID- 21636347 TI - Possible genetic anticipation in families with idiopathic generalised epilepsy. AB - Idiopathic generalised epilepsies (IGE) constitute nearly one third of all epilepsies. IGEs manifest with absences, myoclonic jerks and generalised tonic clonic seizures (GTCS), either alone or in varying combinations, and have a strong genetic background. We present two three-generation families with juvenile myoclonic epilepsy (JME) probands and other affected family members with different forms of IGE in whom genetic anticipation was possible, i.e. the progressive decrease in age at onset with each successive generation. In the first family, the proband presented with JME with all three seizure types with an age at onset of eight years. Her cousin presented with both absence seizures and myoclonic jerks simultaneously at age 14 years, and GTCS occurred one year later. The proband's mother had her first seizures at the age of 39 years (brief myoclonic jerks and subtle absences predated GTCS by a few months). In the second family, the proband and his younger brother presented with JME at the age of 13 years, their mother experienced a single GTCS at the age of 38 years, while the grand-mother died during de novo generalised status at the age of 62 years. To our knowledge, this is one of the few reports to describe the occurrence of possible genetic anticipation in IGE which should be further investigated in larger cohorts of patients. PMID- 21636348 TI - Transcallosal endoscopic resection of hypothalamic hamartoma in a case with Pallister-Hall syndrome. AB - Pallister-Hall syndrome (PHS) is a very rare syndrome characterized by hypothalamic hamartoma (HH), polydactyly, panhypopituitarism, imperforate anus and other visceral anomalies. Contrary to patients with isolated HH, neurological dysfunction and precocious puberty are uncommon and seizures are usually well controlled with anticonvulsant medication. Therefore, conservative management of HH is advised. To the best of our knowledge, seven cases of PHS with surgical resection of the HH have so far been reported. Five patients were either seizure free or had >90% seizure reduction postoperatively. Here, we present a case of PHS of a patient who also underwent transcallosal endoscopic resection of the HH with a subsequent 70% reduction in seizure frequency. PMID- 21636350 TI - Serum interleukin-1 and interleukin-6 are correlated neither with oxidized low density lipoprotein, nor with low-grade inflammation in patients with type 2 diabetes. AB - INTRODUCTION: In vitro studies have shown that oxidized, low density lipoprotein (ox-LDL) stimulates macrophages to release interleukin-6 (IL-6) and interleukin 1beta (IL-1beta). In this study, we aimed to investigate the correlation between ox-LDL and IL-6 and IL-1beta levels in the peripheral circulation of patients with type 2 diabetes, and normal controls. We measured serum high sensitivity CRP (hs-CRP) levels in order to define basal inflammation in patients and controls. METHODS: A total of 40 patients with type 2 diabetes, and 40, age, sex, and body mass index (BMI) -matched healthy adults were enrolled in the study. Fasting blood sugar (FBS), lipid profile, creatinine, ox-LDL, IL-6, IL-1beta and hs-CRP levels were measured. RESULTS: Patients with type 2 diabetes had higher serum FBS, HbA1C, high density lipoprotein cholesterol, LDL, ox-LDL/LDL ratio and hs CRP levels than controls. The higher serum ox-LDL/LDL ratio in patients with type 2 diabetes remained significant after multiple adjustments for age, BMI, FBS and HbA1C, (0.65 [0.59-0.71] vs 0.49 [0.41-0.56]; p<0.001) using general linear models. Serum IL-1beta levels were significantly higher in women than in men with type 2 diabetes; this was not the case in controls. Postmenopausal women in patient and control groups had higher serum IL-6 levels than premenopausal women. There was no significant correlation between serum ox-LDL, ox-LDL/LDL ratio, IL 6, IL-1beta and hs-CRP levels in patients with type 2 diabetes. CONCLUSION: Inflammatory cytokines such as IL-1beta and IL-6 lose their discriminatory power with respect to chronic inflammation in patients with type 2 diabetes. PMID- 21636351 TI - Diagnostic value of resistin and visfatin, in comparison with C-reactive protein, procalcitonin and interleukin-6 in neonatal sepsis. AB - AIM: The aim of this study was to evaluate the predictive value of resistin and visfatin in neonatal sepsis, and to compare these adipocytokines with C-reactive protein (CRP), procalcitonin and interleukin 6 (IL-6). DONORS AND METHODS: A total of 62 term or near term infants with sepsis proven by positivity of blood culture, and 43 healthy infants were included in this study. RESULTS: There were no statistically significant differences between the two groups as regards birthweight and gestational age. White blood cell count (p= 0.039), CRP levels (p=0.01), procalcitonin levels (p=0.01), IL-6 levels (p= 0.01), visfatin levels (p=0.01) and resistin levels (p=0.01) were significantly higher in septic infants. There was a positive correlation between visfatin, resistin and other markers (WBC, CRP, procalcitonin and IL-6). A cut-off value of 10 ng/mL for visfatin, showed 92% sensitivity and 94% specificity, and a cut-off value of 8 ng/mL for resistin showed 93% sensitivity and 95% specificity for neonatal sepsis. CONCLUSION: In the light of these results, visfatin and resistin can be used as a diagnostic marker similar to CRP, procalcitonin and IL-6 in neonatal sepsis. Further studies are needed to better understand the role and predictive value of these molecules in neonatal sepsis. PMID- 21636352 TI - [Use of antioxidant and other complementary medicine by patients treated by antitumor chemotherapy: a prospective study]. AB - Use of complementary and alternative medicine (CAM) has been reported to be more and more frequent among cancer patients in USA. The aim of this study was to analyze among French cancer patients the prevalence of CAM use, focusing on antioxidants (AO) that could interfere with antitumor agents. Seventy-nine patients, treated by antitumor chemotherapy in oncology day care unit, participated to an interview (medium age = 60 years old). CAM use was reported by 42% of patients: mostly AO (24%) (selenium, green tea and vitamins ACE, more specifically), but also relaxation, acupuncture, hypnosis (19%) and homeopathy (15%). Among patients using CAM, 66% of them indicated that their physicians were not aware of this use and 47% of them thought that CAM use was safe. Nevertheless, for seven patients who have taken AO, previous in vitro and preclinical studies suggested interactions with antitumor chemotherapy. Therefore, CAM use and, more specifically, AO use is common among cancer patients treated by antitumor chemotherapy in France. Nevertheless, AO could generate interactions with conventional treatment. Clinical studies are warranted to evaluate these interactions, and adequate communication with patients is needed. PMID- 21636353 TI - Video atlas of lateralising and localising seizure phenomena. AB - The detailed analysis of seizure semiology is an essential tool for diagnosing epileptic patients and is particularly important in the evaluation of patients considered for epilepsy surgery. The meticulous clinical observation of epileptic seizures provides information about the localisation and lateralisation of the symptomatogenic zone. Here, we present a video atlas showing a variety of ictal and postictal localising and lateralising phenomena. [Published with video sequences]. PMID- 21636354 TI - Shaking body attacks: a new type of benign non-epileptic attack in infancy. AB - Non-epileptic attacks represent a heterogeneous group of clinical entities which frequently pose a challenge for the differential diagnosis of epilepsy. This is particularly the case when motor manifestations are the main clinical features. For the large majority of patients, such motor manifestations have a benign course. A correct diagnosis is important to avoid inappropriate investigations, unnecessary therapy, and parental anxiety. Here, a previously unreported form of non-epileptic attacks with infantile onset is described which is different from all subtypes of Fejerman syndrome and does not appear to be uncommon. Our series includes 23 patients with an age at onset of the paroxysmal events ranging from 3 to 8 months. The characteristic feature is side-to-side shaking movements of the trunk and limbs. Surprisingly, urinary infection is often a false diagnosis. Home video recording is particularly helpful in recognising the nature of these episodes once their existence is known. [Published with video sequences]. PMID- 21636355 TI - Recurrent occipital seizures misdiagnosed as status migrainosus. AB - Periictal headache is commonly reported in patients with epilepsy and often exhibits migraine features. Misdiagnosis is frequent since visual seizures may often be misinterpreted as visual aura of migraine. We herein describe a 35-year old woman with recurrent occipital seizures, clinically presenting with intractable headache. EEG monitoring was crucial in order to reach the correct diagnosis. PMID- 21636356 TI - Infliximab-related seizures: a first case study. AB - Seizures following infliximab treatment are very rare and, to date, there is no detailed description of EEG abnormalities with cerebral radiological findings reported in cases with infliximab-related seizures. We describe a patient who acutely developed seizures temporally related to infliximab treatment, which disappeared after drug withdrawal. MRI showed encephalopathy involving mainly cortical regions and EEGs showed focal paroxysmal activity which completely disappeared a few days after infliximab withdrawal. No other plausible cause of the seizures was identified. The clear temporal association between seizure onset and infliximab treatment as well as the clinical improvement and disappearance of focal epileptiform activity after drug withdrawal indicated an evident correlation between seizures and infliximab therapy. The coexistence of pathological findings on MRI suggested that seizures were secondary to the encephalopathy. Further studies are required to evaluate whether infliximab per se has an epileptogenic effect or whether the seizures are caused by encephalopathy involving cortico-subcortical regions. PMID- 21636357 TI - How to determine sapling buckling risk with only a few measurements. AB - Tree buckling risk (actual height/critical buckling height) is an important biomechanical trait of plant growth strategies, and one that contributes to species coexistence. To estimate the diversity of this trait among wide samples, a method that minimizes damage to the plants is necessary. On the basis of the rarely used, complete version of Greenhill's model (1881, Proceedings of the Cambridge Philosophical Society 4(2): 65-73), we precisely measured all the necessary parameters on a sample of 236 saplings of 16 species. Then, using sensitivity (variance) analysis, regressions between successive models for risk factors and species ranks and the use of these models on samples of self- and nonself-supporting saplings, we tested different degrees of simplification up to the most simple and widely used formula that assumes that the tree is a cylindrical homogeneous pole. The size factor had the greatest effect on buckling risk, followed by the form factor and the modulus of elasticity of the wood. Therefore, estimates of buckling risk must consider not only the wood properties but especially the form factor. Finally, we proposed a simple but accurate method of assessing tree buckling risk that is applicable to a wide range of samples and that requires mostly nondestructive measurements. PMID- 21636358 TI - Density and seed set in a self-compatible forb, Penstemon digitalis (Plantaginaceae), with multiple pollinators. AB - Seed production may be limited because flowers do not get enough suitable pollen or because plants lack the resources to make seeds. We used replicated plantings to test factors that influence effects of bumblebee behavior on pollen limitation, as measured by the difference in seed set between hand- and naturally pollinated flowers, of Penstemon digitalis in patches of four to 41 flowering individuals. Seed set per flower was 376% higher in the largest as compared with the smallest Penstemon patches. This positive density dependence reflects activity of long-tongued bees, which (1) have higher effective density as patch size increases, (2) visit greater proportions of plants as patch size increases, and (3) visit smaller proportions of flowers per visited plant as patch size increases. Our results suggest that economics of flight and maneuverability of large, long-tongued bumblebees lead them to transfer more pollen between than within Penstemon plants in large patches. Density of smaller, short-tongued bumblebees was not positively associated with Penstemon seed set, but these bees may be important pollinators at low plant densities. Our experimental system indicates a clear positive relationship between activity of effective pollinators and seed set in a species capable of pollinating itself. PMID- 21636359 TI - Multiple resonance damping or how do trees escape dangerously large oscillations? AB - To further understand the mechanics of trees under dynamic loads, we recorded damped oscillations of a Douglas fir (Pseudotsuga menziesii) tree and of its stem without branches. Eigenfrequencies of the branches were calculated and compared to the oscillation frequency of the intact tree. The term eigenfrequency is used here to characterize the calculated resonance frequency of a branch fixed at the proximal end to a solid support. All large branches had nearly the same frequency as the tree. This property is a prerequisite for the distribution of mechanical energy between stem and branches and leads to an enhanced efficiency of damping. We propose that trees constitute systems of coupled oscillators tuned to allow optimal energy dissipation. PMID- 21636360 TI - Cryptic species and host specificity in the ectomycorrhizal genus Strobilomyces (Strobilomycetaceae). AB - Taxonomical classification of higher fungi remains an important challenge and can benefit from the application of molecular analysis. We propose that the ectomycorrhizal (EM) fungal taxa might include a number of cryptic species because there are few morphological characteristics useful for distinguishing among these fungi. Previously, host specificity in most EM fungi was thought to be low, but we suspect that confusion of cryptic species has led to an underestimate of fungal host specificity. We analyzed both nuclear and mitochondrial DNA sequences from Strobilomyces fungi and obtained evidence that what were previously described as four species can be grouped into 14 distinct lineages, suggesting that these lineages might be distinct biological species. Moreover, we identified host plants for Strobilomyces via nucleotide sequencing of both fungal and plant DNA from EM samples. Most lineages of Strobilomyces tested in this study were associated only with Fagaceae trees, even though Strobilomyces species were previously thought to be generalists with regard to hosts. Thus, we present an approach useful for identifying cryptic species and detecting the true host range of a set of EM fungi in natural conditions. PMID- 21636361 TI - Establishing a fossil record for the perianthless Piperales: Saururus tuckerae sp. nov. (Saururaceae) from the Middle Eocene Princeton Chert. AB - Investigations of small permineralized flowers from the Middle Eocene Princeton Chert, British Columbia, Canada have revealed that they represent an extinct species of Saururus. Over 100 flowers and one partial inflorescence were studied, and numerous minute perianthless flowers are borne in an indeterminate raceme. Each flower is subtended by a bract, and flowers and bracts are borne at the end of a common stalk. Five stamens are basally adnate to the carpels. Pollen is frequently found in situ in the anthers. Examined under SEM and TEM, pollen grains are minute (6-11 MUm), monosulcate, boat-shaped-elliptic, with punctate sculpturing and a granulate aperture membrane. The gynoecium is composed of four basally connate, lobed carpels with recurved styles and a single ovule per carpel. Flower structure and pollen are indicative of Saururaceae (Piperales), and in phylogenetic analyses using morphological characters, the fossils are sister to extant Saururus. The fossil flowers are described here as Saururus tuckerae sp. nov. These fossil specimens add to the otherwise sparse fossil record of Piperales, represent the oldest fossils of Saururaceae as well as the first North American fossil specimens of this family, and provide the first evidence of saururaceous pollen in the fossil record. PMID- 21636362 TI - Reproductive structures of Rhamnaceae from the Cerro del Pueblo (Late Cretaceous, Coahuila) and Coatzingo (Oligocene, Puebla) Formations, Mexico. AB - Recently discovered fossil flowers from the Cretaceous Cerro del Pueblo and flowers and fruits from the Oligocene Coatzingo Formations are assigned to the Rhamnaceae. The Cretaceous flower, Coahuilanthus belindae Calvillo-Canadell and Cevallos-Ferriz, gen. et sp. nov., is actinomorphic with fused perianth parts forming a slightly campanulate to cupulate floral cup, with sepals slightly keeled and spatulate clawed petals. The Oligocene fossils include Nahinda axamilpensis Calvillo-Canadell and Cevallos-Ferriz, gen. et sp. nov. (characterized by its campanulate bisexual flower with stamens opposite, adnate to and enfolded by petals; and with the ovary ripening into a drupe), and a winged fruit assigned to Ventilago engoto Calvillo-Canadell and Cevallos-Ferriz, sp. nov. The flowers and drupe features indicate closer affinity to Zizipheae and/or Rhamneae, while the single samaroid fruit suggests the presence of Ventilagineae. However, the unique character combination in the fossil flowers precludes placing them in extant genera. Nevertheless, the history of the family is long and can be traced back to the Campanian. A detailed phylogenetic revision of the group that uses morphological characters from both extant and fossil plants is needed to better understand the significance of these records as well as other important fossils of the family. PMID- 21636363 TI - Crassulacean acid metabolism in the ZZ plant, Zamioculcas zamiifolia (Araceae). AB - Zamioculcas zamiifolia (Araceae), a terrestrial East African aroid, with two defining attributes of crassulacean acid metabolism (CAM) (net CO(2) uptake in the dark and diel fluctuations of titratable acidity) is the only CAM plant described within the Araceae, a mainly tropical taxon that contains the second largest number of epiphytes of any vascular plant family. Within the Alismatales, the order to which the Araceae belong, Z. zamiifolia is the only documented nonaquatic CAM species. Zamioculcas zamiifolia has weak CAM that is upregulated in response to water stress. In well-watered plants, day-night fluctuations in titratable acidity were 2.5 MUmol H(+).(g fresh mass)(-1), and net CO(2) uptake in the dark contributed less than 1% to daily carbon gain. Following 10 d of water stress, net CO(2) uptake in the light fell 94% and net CO(2) uptake in the dark increased 7.5-fold, such that its contribution increased to 19% of daily carbon gain. Following rewatering, dark CO(2) uptake returned to within 5% of prestressed levels. We postulate that CAM assists survival of Z. zamiifolia by reducing water loss and maintaining carbon gain during seasonal droughts characteristic of its natural habitat. PMID- 21636364 TI - Effects of self-pollination and outcrossing with cultivated plants in small natural populations of American ginseng, Panax quinquefolius (Araliaceae). AB - For rare plants, self-pollination and inbreeding can increase in small populations, while unusual levels of outcrossing can occur through restoration efforts. To study both inbreeding and outcrossing, we performed experimental pollinations using Panax quinquefolius (American ginseng), a wild-harvested plant with a mixed mating system. For inbreeding, plants were either cross-pollinated within the population or self-pollinated, which resulted in a higher proportion of seeds from self-pollinated flowers. For outcrossing, wild plants were either cross-pollinated within the population or with cultivated plants from West Virginia or Wisconsin. Offspring of all crosses were followed for 4 yr. Two-yr old seedlings from self-pollination had 45% smaller leaf areas and 33% smaller heights relative to those from cross-pollination. Leaf area is a positive predictor of longer-term survival in wild populations. Our results suggest inbreeding depression, which is unexpected in this self-fertile species. Seedlings from crosses with cultivated plants had 127% greater leaf area and 165% greater root biomass relative to outcrosses within the population. The accelerated growth suggests genetic differences between wild and cultivated populations, but outbreeding depression may not appear until later generations. Assessment of the ultimate fitness consequences of introducing cultivated genotypes requires monitoring over longer time periods. PMID- 21636365 TI - The impact of pollination syndrome and habitat on gene flow: a comparative study of two Streptocarpus (Gesneriaceae) species. AB - Gene flow through pollen and seed dispersal is important in terms of population differentiation and eventually speciation. Seed and pollen flow are affected in turn by habitats and pollen vectors. We examined the effect of different pollinators and habitats on gene flow by comparing two species of Streptocarpus, using microsatellite and chloroplast RFLP markers. Populations of the forest dwelling S. primulifolius were highly differentiated according to nuclear microsatellite data and had mutually exclusive chloroplast haplotypes. This result is congruent with infrequent seed dispersal and limited between-population foraging by the long-tongued fly pollinator Stenobasipteron wiedemanni. In contrast, populations of S. dunnii growing in exposed crags had lower levels of population differentiation according to both nuclear and chloroplast data, congruent with a hypothesis of more effective between population seed dispersal and greater pollen-mediated gene flow due to the sunbird pollinator Nectarinia famosa. The population genetic behavior of these species is reflected in their taxonomy and phylogenetic position; S. primulifolius belongs to a taxonomically complex clade in which recent speciation is evident, while the clade containing S. dunnii is characterized by taxonomically well-defined species on longer phylogenetic branches. Our study shows that pollinator movements and seed dispersal patterns are a major determinant of the evolutionary trajectories of these species. PMID- 21636366 TI - Effects of floral display size and biparental inbreeding on outcrossing rates in Delphinium barbeyi (Ranunculaceae). AB - Floral display size represents a tradeoff between the benefits of increased pollinator visitation and the quantity of pollen received vs. the costs of increased self-pollination and reduced pollination quality. Plants with large floral displays often are more attractive to pollinators, but pollinators visit more flowers per plant. Intraplant foraging movements should increase self pollination through geitonogamy, lowering outcrossing rates in large plants. Local genetic structure should also increase inbreeding and decrease outcrossing estimates, if pollinators move between neighboring, related plants. These predictions were tested in a population of larkspurs (Delphinium barbeyi) in Colorado. Allozymes were used to estimate outcrossing rates of plants varying in display size. Floral displays varied widely (2-1400 flowers; 1-26 inflorescences per plant), and outcrossing rate decreased significantly with increasing display size. Large, multistalked plants self over twice as frequently as single-stalked plants (46 vs. 21%). Local population structure is significant, and biparental inbreeding depresses outcrossing in plants surrounded by genetically similar neighbors. Protandry, coupled with stereotypical bottom-up pollinator foraging, reduces self-fertilization by autogamy or geitonogamy within inflorescences. Selfing is predominantly (>60%) by geitonogamy between inflorescences in large plants. Geitonogamy may be a significant cost to plants with large floral displays if inbreeding depression and/or pollen and ovule discounting results. If so, floral display size, particularly inflorescence number, may be under contrasting selection for pollination quantity vs. quality. PMID- 21636367 TI - Correlated evolution of fruit size and sexual expression in andromonoecious Solanum sections Acanthophora and Lasiocarpa (Solanaceae). AB - Andromonoecy is hypothesized to evolve as a mechanism enabling plants to independently allocate resources to female and male function. If staminate flower production is a mechanism to regulate allocation to female function (i.e., fruit production), then large-fruited species should be more strongly andromonoecious than smaller-fruited taxa because more resources are required to mature large fruit. We combined phylogenetically independent contrast analyses with extensive phenotypic characterization under common greenhouse conditions to examine the predicted relationship between fruit mass and the strength of andromonoecy among 13 species in Solanum sections Acanthophora and Lasiocarpa. The strength of andromonoecy, defined as the proportion of staminate flowers produced within inflorescences, was significantly and positively associated with fruit mass in both naive and phylogenetically independent analyses. Our results are consistent with the hypothesis that andromonoecy functions as a mechanism to regulate allocation to female function and suggest that the strength of andromonoecy is also associated with resource limitation. In general, we find that strong andromonoecy appears to arise via reductions in hermaphroditic flower number. However, increases in staminate flowers have also contributed to transitions to strong andromonoecy in certain species. Finally, our analyses identified a suite of correlated characters (flower size, ovary width, fruit mass) that are associated with changes in the sexual expression of andromonoecy. PMID- 21636368 TI - Sexually dimorphic inflorescence traits in a wind-pollinated species: heritabilities and genetic correlations in Schiedea adamantis (Caryophyllaceae). AB - Sexual dimorphism may be especially pronounced in wind-pollinated species because they lack the constraints of biotically pollinated species that must present their pollen and stigmas in similar positions to ensure pollen transfer. Lacking these constraints, the sexes of wind-pollinated species may diverge in response to the different demands of pollen dispersal and receipt, depending on the magnitude of genetic correlations preventing divergence between sexes. Patterns of sexual dimorphism and genetic variation were investigated for inflorescence traits in Schiedea adamantis (Caryophyllaceae), a species well adapted to wind pollination, and compared to S. salicaria, a species with fewer adaptations to wind pollination. For S. adamantis, dimorphism was pronounced for inflorescence condensation and its components, including lateral flower number and pedicel length. Within sexes, genetic correlations between traits may constrain the relative shape of the inflorescence. Correlations detected across sexes may retard the evolution of sexual dimorphism in inflorescence structure, including features favoring enhanced dispersal and receipt of pollen. Despite genetic correlations across sexes, common principal components analysis showed that genetic variance-covariance matrices (G matrices) differed significantly between the sexes, in part because of greater genetic variation for flower number in hermaphrodites than in females. G matrices also differed between closely related S. adamantis and S. salicaria, indicating the potential for divergent evolution of inflorescence structure despite general similarities in morphology and pollination biology. PMID- 21636369 TI - Molecular phylogeny of Macaranga, Mallotus, and related genera (Euphorbiaceae s.s.): insights from plastid and nuclear DNA sequence data. AB - Macaranga and Mallotus (Euphorbiaceae s.s.) are two closely related, large paleo(sub)tropical genera. To investigate the phylogenetic relationships between and within them and to determine the position of related genera belonging to the subtribe Rottlerinae, we sequenced one plastid (trnL-F) and three nuclear (ITS, ncpGS, phyC) markers for species representative of these genera. The analyses demonstrated the monophyly of Macaranga and the paraphyly of Mallotus and revealed three highly supported main clades. The genera Cordemoya and Deuteromallotus and the Mallotus sections Hancea and Oliganthae form a basal Cordemoya s.l. clade. The two other clades, the Macaranga clade and the Mallotus s.s. clade (the latter with Coccoceras, Neotrewia, Octospermum, and Trewia), are sister groups. In the Macaranga clade, two basal lineages (comprising mostly sect. Pseudorottlera) and a crown group with three geographically homogenous main clades were identified. The phylogeny of the Mallotus s.s. clade is less clear because of internal conflict in all four data sets. Many of the sections and informal infrageneric groups of Macaranga and Mallotus do not appear to be monophyletic. In both the Macaranga and Mallotus s.s. clades, the African and/or Madagascan taxa are nested in Asian clades, suggesting migrations or dispersals from Asia to Africa and Madagascar. PMID- 21636370 TI - The role of auxin transport during inflorescence development in maize (Zea mays, Poaceae). AB - Axillary meristems play a fundamental role in inflorescence architecture. Maize (Zea mays) inflorescences are highly branched panicles because of the production of multiple types of axillary meristems. We used auxin transport inhibitors to show that auxin transport is required for axillary meristem initiation in the maize inflorescence. The phenotype of plants treated with auxin transport inhibitors is very similar to that of barren inflorescence2 (bif2) and barren stalk1 (ba1) mutants, suggesting that these genes function in the same auxin transport pathway. To dissect this pathway, we performed RNA in situ hybridization on plants treated with auxin transport inhibitors. We determined that bif2 is expressed upstream and that ba1 is expressed downstream of auxin transport, enabling us to integrate the genetic and hormonal control of axillary meristem initiation. In addition, treatment of maize inflorescences with auxin transport inhibitors later in development results in the production of single instead of paired spikelets. Paired spikelets are a key feature of the Andropogoneae, a group of over 1000 grasses that includes maize, sorghum, and sugarcane. Because all other grasses bear spikelets singly, these results implicate auxin transport in the evolution of inflorescence architecture. Furthermore, our results provide insight into mechanisms of inflorescence branching that are relevant to all plants. PMID- 21636371 TI - Glomeromycotean associations in liverworts: a molecular, cellular, and taxonomic analysis. AB - Liverworts form endophytic associations with fungi that mirror mycorrhizal associations in tracheophytes. Here we report a worldwide survey of liverwort associations with glomeromycotean fungi (GAs), together with a comparative molecular and cellular analysis in representative species. Liverwort GAs are circumscribed by a basal assemblage embracing the Haplomitriopsida, the Marchantiopsida (except a few mostly derived clades), and part of the Metzgeriidae. Fungal endophytes from Haplomitrium, Conocephalum, Fossombronia, and Pellia were related to Glomus Group A, while the endophyte from Monoclea was related to Acaulospora. An isolate of G. mosseae colonized axenic thalli of Conocephalum, producing an association similar to that in the wild. Fungal colonization in marchantialean liverworts suppressed cell wall autofluorescence and elicited the deposition of a new wall layer that specifically bound the monoclonal antibody CCRC-M1 against fucosylated side groups associated with xyloglucan and rhamnogalacturonan I. The interfacial material covering the intracellular fungus contained the same epitopes present in host cell walls. The taxonomic distribution and cytology of liverwort GAs suggest an ancient origin and multiple more recent losses, but the occurence in widely separated liverwort taxa of fungi related to glomeromycotean lineages that form arbuscular mycorrhizas in tracheophytes, notably the Glomus Group A, is better explained by host shifting from tracheophytes to liverworts. PMID- 21636372 TI - Complementarity in mineral nitrogen use among dominant plant species in a subalpine community. AB - The underlying mechanisms that enable plant species to coexist are poorly understood. Complementarity in resource use is among the major mechanisms proposed that could favor species coexistence but is insufficiently documented. In alpine soil, low temperatures are a major constraint for the supply of plant nitrogen. We carried out (15)N labeling of soil mineral N to determine to what extent four major species of a subalpine community compete for N, or develop ionic (NH(4)(+) vs. NO(3)(-)) or temporal complementarity. The Poaceae took up much more (15)N per soil area unit than the ericaceous species, and all species displayed three major strategies in exploiting (15)N: (1) uptake mainly early in the growing season (Vaccinium myrtillus), (2) uptake at a slow and similar rate throughout the growing season (Rhododendron ferrugineum), and (3) uptake at high rates over the growing season (Festuca eskia and Nardus stricta). However, while F. eskia used (15)NH(4)(+) mainly early and (15)NO(3)(-) mainly late in the growing season, the reverse was observed for N. stricta. Taking into account (15)N dilution in soil NH(4)(+) and NO(3)(-) pools, we calculated that NH(4)(+) provided more than 80% of the mineral N uptake in Ericaceae and about 60% in grasses. Together, such ionic and temporal complementarity would reduce competition between species and could be a major mechanism promoting species diversity. PMID- 21636373 TI - Estimation of gene flow into fragmented populations of Bursera simaruba (Burseraceae) in the dry-forest life zone of Puerto Rico. AB - We examined the impact of habitat fragmentation on gene flow in populations of the neotropical tree Bursera simaruba. In particular, we compared the effectiveness of three common techniques to estimate gene flow in the context of a highly disturbed system. Paternity analysis on emerging seedlings from eight small (N = 3 to 9) stands of trees showed that between 45% and 100% of seedlings were sired from outside their stand, indicating pollen moved readily over the isolation distances examined. Based on six populations of 21-24 trees each, estimates of allozyme genetic diversity (P(s) = 73.3%; H(e) = 0.244) were higher than those reported for species with similar life history traits. Indirect, F(ST) based gene flow estimates for these six populations yielded an estimate of 3.57 migrants per generation, although possible violations of model assumptions limit the reliability of the estimate. A twogener analysis showed pollen moved either 320 m or 361 m and that there were only 2.46 effective pollen donors per maternal tree. Despite the potential for long-distance pollen movement, seed abortion was high, especially in stands with fewer than four trees. Population size, rather than isolation distance, appears to limit reproduction in the populations examined. PMID- 21636374 TI - Seed-mass effects in four Mediterranean Quercus species (Fagaceae) growing in contrasting light environments. AB - Three hypotheses have been proposed to explain the functional relationship between seed mass and seedling performance: the reserve effect (larger seeds retain a larger proportion of reserves after germinating), the metabolic effect (seedlings from larger seeds have slower relative growth rates), and the seedling size effect (larger seeds produce larger seedlings). We tested these hypotheses by growing four Mediterranean Quercus species under different light conditions (3, 27, and 100% of available radiation). We found evidence for two of the three hypotheses, but none of the four species complied with all three hypotheses at the same time. The reserve effect was not found in any species, the metabolic effect was found in three species (Q. ilex, Q. pyrenaica, and Q. suber), and the seedling-size effect in all species. Light availability significantly affected the relationships between seed size and seedling traits. For Q. ilex and Q. canariensis, a seedling-size effect was found under all three light conditions, but only under the lowest light (3%) for Q. suber and Q. pyrenaica. In all species, the correlation between seed mass and seedling mass increased with a decrease in light, suggesting that seedlings growing in low light depend more upon their seed reserves. A causal model integrates the three hypotheses, suggesting that larger seeds generally produced larger seedlings. PMID- 21636375 TI - Leaf physiology reflects environmental differences and cytoplasmic background in Ipomopsis (Polemoniaceae) hybrids. AB - Natural hybridization can produce individuals that vary widely in fitness, depending upon the performance of particular genotypes in a given environment. In a hybrid zone with habitat heterogeneity, differences in physiological responses to abiotic conditions could influence the fitness and spatial distribution of hybrids and parental species. This study compared gas exchange physiology of Ipomopsis aggregata, I. tenuituba, and their natural hybrids in situ and assessed whether physiological differences were consistent with their native environmental conditions. We also produced reciprocal F2s in a greenhouse study to test for cytonuclear effects on water-use efficiency (WUE). The relative performance of natural hybrids and parentals was consistent with their native habitats: I. aggregata at the coolest, wettest locations had the lowest WUE, while hybrids from the most xeric sites had the highest WUE. In hybrids, the mechanism by which both natural and experimental hybrids achieved this high WUE depended on cytotype: those with I. tenuituba cytoplasm had reduced transpiration, while those with I. aggregata cytoplasm had an increased photosynthetic rate, consistent with patterns in the cytoplasmic parent. The high WUE in hybrids may contribute to their high survival in the dry center of the natural hybrid zone, consistent with environment-dependent models of hybrid zone dynamics. PMID- 21636376 TI - Infection by powdery mildew Erysiphe cruciferarum (Erysiphaceae) strongly affects growth and fitness of Alliaria petiolata (Brassicaceae). AB - Garlic mustard (Alliaria petiolata) is an invasive biennial that negatively impacts plant and animal communities throughout North America and lacks significant herbivory in its invasive range. Throughout Ohio, many garlic mustard populations support the powdery mildew fungus Erysiphe cruciferarum, although disease incidence varies among populations and environments. Effects of infection on plant growth, as well as both plant and fungal responses to drought and light conditions, were examined on greenhouse-grown, first-year garlic mustard plants. Also, the effects of the fungus on plant growth and fitness were studied in a naturally growing population of second-year plants in the field. Powdery mildew significantly reduced growth of first-year plants in the greenhouse, eventually causing complete mortality. Simulated drought slowed both plant growth and disease development, independent of light conditions. In the field, plants with little incidence of disease after their first year grew taller during their second year, producing significantly more siliques and twice as many seeds as heavily diseased plants did. Seed germination rates did not differ between plants with different levels of disease severity. Consistent reductions in survival, growth, and fitness caused by fungal infection may reduce populations of garlic mustard. These effects may be more evident in moist sites that favor fungal development. PMID- 21636377 TI - Swietenia (Meliaceae) flower in Late Oligocene Early Miocene amber from Simojovel de Allende, Chiapas, Mexico. AB - The amber of Simojovel de Allende, Chiapas, Mexico, of Late Oligocene-Early Miocene age, has yielded a new flower representing the Meliaceae. The flower of Swietenia miocenica Castaneda-Posadas & Cevallos-Ferriz sp. nov. is characterized by small size; free calyx composed of five glabrous lobes, ciliolated along the margin lobes; corolla composed of five free, contortedly inserted petals with ciliolated margins; cylindrical staminal tube ending in 10 acuminate or toothed accessories and 10 sessile anthers; and a discoid stigma divided in five lobular stigmatic glands. The morphology of S. miocenica is well represented among Meliaceae. Although the new species shares many characters with Swietenia microphylla, small differences in the length and width of petals and the length of staminal tube support its recognition as a new species. The presence of this genus demonstrates the establishment of tropical communities in southern Mexico by the early Miocene and highlights the influence of the northern hemisphere flora on the extant neotropical flora of the area. PMID- 21636378 TI - Flower development of Meliosma (Sabiaceae): evidence for multiple origins of pentamery in the eudicots. AB - Flower developmental studies are a complement to molecular phylogenetics and a tool to understand the evolution of the angiosperm flower. Buds and mature flowers of Meliosma veitchiorum, M. cuneifolia, and M. dilleniifolia (Sabiaceae) were investigated using scanning electron microscopy to clarify flower developmental patterns and morphology, to understand the origin of the perianth merism, and to discuss the two taxonomic positions proposed for Sabiaceae, among rosids or in the basal grade of eudicots. Flowers in Meliosma appear pentamerous with two of the five sepals and petals strongly reduced, three staminodes alternating with two fertile stamens opposite the small petals, and a two carpellate gynoecium. The flower development in Meliosma is spiral without distinction between bracteoles and sepals. Because of this development, sepals, petals, and stamens are almost opposite and not alternating as expected in cyclical pentamerous flowers. In four-sepal flowers the direction of petal initiation is reversed. The symmetry of the flower appears to be transversally zygomorphic, although this is hidden by the almost equal size of the larger petals. Evidence points to a unique pentamerous origin of flowers in Meliosma, and not to a trimerous origin, as earlier suggested, and adds support to multiple origins of pentamery in the eudicots. PMID- 21636379 TI - High-resolution phylogeny for Helianthus (Asteraceae) using the 18S-26S ribosomal DNA external transcribed spacer. AB - The sunflower genus, Helianthus, is recognized widely for the cultivated sunflower H. annuus and scientifically as a model organism for studying diploid and polyploid hybrid speciation, introgression, and genetic architecture. A resolved phylogeny for the genus is essential for the advancement of these scientific areas. In the past, phylogenetic relationships of the perennial species and polyploid hybrids have been particularly difficult to resolve. Using the external transcribed spacer region of the nuclear 18S-26S rDNA region, we reveal for the first time a highly resolved gene tree for Helianthus. Phylogenetic analysis allowed the determination of a monophyletic annual H. sect. Helianthus, a two-lineage polyphyletic H. sect. Ciliares, and the monotypic H. sect. Agrestis, all of which were nested within a large perennial and polyphyletic H. sect. Divaricati. The distribution of perennial polyploids and known annual diploid hybrids on this phylogeny suggested multiple independent hybrid speciation events that gave rise to at least four polyploids and three diploid hybrids. Also provided by this phylogeny was evidence for homoploid hybrid speciation outside H. sect. Helianthus. Finally, previous hypotheses about the secondary chemistry in the genus were tested in a phylogenetic framework to obtain a better understanding of the evolution of these compounds in Helianthus. PMID- 21636380 TI - Sporophytic inbreeding depression in mosses occurs in a species with separate sexes but not in a species with combined sexes. AB - Inbreeding depression is a critical factor countering the evolution of inbreeding and thus potentially shaping the evolution of plant sexual systems. Current theory predicts that inbreeding depression could have important evolutionary consequences, even in haploid-dominant organisms. To date, no data have been reported on inbreeding depression in moss species. Here, we present data on the magnitude of inbreeding depression in sporophytic traits of moss species with contrasting breeding systems. In Ceratodon purpureus (Ditrichaceae), a moss species with separate sexes, self-fertilizations between sibling gametophytes (intergametophytic selfing) significantly reduced fitness in two of four traits quantified, with seta length and capsule length having inbreeding coefficients significantly different from zero, resulting in a cumulative inbreeding depression that was also significantly greater than zero (delta = 0.619 +/- 0.076). In hermaphroditic Funaria hygrometrica (Funariaceae), there was no evidence of inbreeding depression in seta length, spore number, capsule mass, or capsule length resulting from sporophytes generated by self-fertilization within an individual (intragametophytic selfing), and cumulative inbreeding depression was also not different from zero (delta = 0.038 +/- 0.022). These results provide evidence that, despite haploid dominance, inbreeding depression can be expressed at the diploid stage in mosses and may have implications for the evolution and maintenance of combined versus separate sexes in mosses. PMID- 21636381 TI - Molecular phylogenetics of Maxillaria and related genera (Orchidaceae: Cymbidieae) based on combined molecular data sets. AB - The orchid genus Maxillaria is one of the largest and most common of neotropical orchid genera, but its current generic boundaries and relationships have long been regarded as artificial. Phylogenetic relationships within subtribe Maxillariinae sensu Dressler (1993) with emphasis on Maxillaria s.l. were inferred using parsimony analyses of individual and combined DNA sequence data. We analyzed a combined matrix of nrITS DNA, the plastid matK gene and flanking trnK intron, and the plastid atpB-rbcL intergenic spacer for 619 individuals representing ca. 354 species. The plastid rpoC1 gene (ca. 2600 bp) was sequenced for 84 selected species and combined in a more limited analysis with the other data sets to provide greater resolution. In a well-resolved, supported consensus, most clades were present in more than one individual analysis. All the currently recognized minor genera of "core" Maxillariinae (Anthosiphon, Chrysocycnis, Cryptocentrum, Cyrtidiorchis, Mormolyca, Pityphyllum, and Trigonidium) are embedded within a polyphyletic Maxillaria s.l. Our results support the recognition of a more restricted Maxillaria, of some previously published segregate genera (Brasiliorchis, Camaridium, Christensonella, Heterotaxis, Ornithidium, Sauvetrea), and of several novel clades at the generic level. These revised monophyletic generic concepts should minimize further nomenclatural changes, encourage monographic studies, and facilitate more focused analyses of character evolution within Maxillariinae. PMID- 21636382 TI - Ascoma development and phylogeny of an apothecioid dothideomycete, Catinella olivacea. AB - Catinella olivacea is a discomycetous fungus often found fruiting within cavities in rotting logs. Because this habitat would lack the air currents upon which discomycete species normally rely for the dispersal of their forcibly ejected ascospores, we suspected an alternative disseminative strategy might be employed by this species. An examination of the development of the discomycetous ascomata in pure culture, on wood blocks, and on agar showed that the epithecium was gelatinous at maturity and entrapped released ascospores in a slimy mass. We interpreted this as an adaptation for ascospore disperal by arthropods. Developmental data also showed that C. olivacea was unusual among other discomycetes in the Helotiales (Leotiomycetes). For example, the ascoma developed from a stromatic mass of meristematically dividing cells and involved the formation of a uniloculate cavity within a structure better considered an ascostroma than an incipient apothecium. Furthermore, the ascus had a prominent ocular chamber and released its ascospores through a broad, bivalvate slit. These features, along with phylogenetic analyses of large subunit and small subunit rDNA, indicated that this unusual apothecial fungus is, surprisingly, more closely affiliated with the Dothideomycetes than the Leotiomycetes. PMID- 21636383 TI - Hybridization and morphogenetic variation in the invasive alien Fallopia (Polygonaceae) complex in Belgium. AB - The invasive alien knotweeds, Fallopia spp. (Polygonaceae), are some of the most troublesome invasive species in Europe and North America. Invasive success in Fallopia may be enhanced by multiple hybridization events. We examined the pattern of hybridization and its evolutionary consequences in Belgium with a concerted analysis of ploidy levels (chromosome counts and flow cytometry), morphological variation, and genetic variation (RAPDs). At least four taxa with different ploidy levels were part of the pattern of invasion in Belgium. Hybrid F. *bohemica with various chromosome numbers restored the genotypic diversity that was lacking in the parental species. Hybrid genotypes were mainly assigned to a specific genetic pool and not to a mixture between the genetic pools of the putative parental species as would be expected for hybrids. Parental species and hexaploid hybrids differed significantly for a set of well-defined morphological characters, enabling future researchers to distinguish these taxa. On the basis of our results, the importance of hybridization has probably been underestimated in large parts of the adventive range of alien Fallopia species, pointing to the need for concerted molecular and morphological analyses in the study of the evolutionary consequences of hybridization. PMID- 21636384 TI - Errata. PMID- 21636385 TI - Evolution of shoot apical meristem structures in vascular plants with respect to plasmodesmatal network. AB - Vascular plants have evolved shoot apical meristems (SAMs), whose structures differ among plant groups. To clarify the evolutionary course of the different structural types of SAMs, we compared plasmodesmatal networks in the SAMs for 17 families and 24 species of angiosperms, gymnosperms, and pteridophytes, using transmission electron microscopy (TEM). The plasmodesmata (PD) in almost all cell walls in median longitudinal sections of SAMs were counted, and the PD density per unit area was calculated for each cell wall. Angiosperm and gymnosperm SAMs have low densities, with no difference between stratified (tunica-corpus) and unstratified structures. SAMs of ferns, including Psilotum and Equisetum, have average densities that are more than three times higher than those of seed plants. Interestingly, microphyllous lycopods have both the fern and seed-plant types of PD networks; Selaginellaceae SAMs with single apical cells have high PD densities, while SAMs of Lycopodiaceae and Isoetaceae with plural initial cells have low PD densities, equivalent to those of seed plants. In summary, PD networks are strongly correlated to SAM organizations-SAMs with single and plural initial cells have the fern and seed-plant types of PD, respectively. The two SAM organizations may have evolved separately in lycophytes and euphyllophytes and may be associated with gain or loss of the ability to form secondary PD. PMID- 21636386 TI - Bizonoplast, a unique chloroplast in the epidermal cells of microphylls in the shade plant Selaginella erythropus (Selaginellaceae). AB - Study of the unique leaf anatomy and chloroplast structure in shade-adapted plants will aid our understanding of how plants use light efficiently in low light environments. Unusual chloroplasts in terms of size and thylakoid membrane stacking have been described previously in several deep-shade plants. In this study, a single giant cup-shaped chloroplast, termed a bizonoplast, was found in the abaxial epidermal cells of the dorsal microphylls and the adaxial epidermal cells of the ventral microphylls in the deep-shade spike moss Selaginella erythropus. Bizonoplasts are dimorphic in ultrastructure: the upper zone is occupied by numerous layers of 2-4 stacked thylakoid membranes while the lower zone contains both unstacked stromal thylakoids and thylakoid lamellae stacked in normal grana structure oriented in different directions. In contrast, other cell types in the microphylls contain chloroplasts with typical structure. This unique chloroplast has not been reported from any other species. The enlargement of epidermal cells into funnel-shaped, photosynthetic cells coupled with specific localization of a large bizonoplast in the lower part of the cells and differential modification in ultrastructure within the chloroplast may allow the plant to better adapt to low light. Further experiments are required to determine whether this shade-adapted organism derives any evolutionary or ecophysiological fitness from these unique chloroplasts. PMID- 21636387 TI - Anisotropic viscosity of the Chara (Characeae) rhizoid cytoplasm. AB - To characterize cellular fluidity and mechanical processes, we determined the viscous properties of the cytoplasm of Chara contraria rhizoids in vivo by injecting and displacing superparamagnetic particles. After injection and a 24-h recovery period, the particles were moved to different positions within the rhizoid by an external magnet. The system was calibrated with solutions of known viscosities. The viscosity was determined based on the velocity at which individual beads moved toward the external magnet. The viscosity of the cytoplasm varied with direction of measurement (i.e., was highly anisotropic) and also varied between sites. The highest viscosity was observed near the endogenous statoliths (139 mP.s parallel and 78 mP.s perpendicular to the rhizoid axis). Depolymerization of actin filaments with latrunculin B reduced the viscosity significantly except around the nucleus but did not change the overall viscosity pattern. Microtubule depolymerization with oryzalin reduced viscosity especially between the nucleus and the statolith zone. The data indicate that F-actin but not microtubules affects statolith sedimentation and that cytoplasmic viscosity may be important for the gravisensing system. PMID- 21636388 TI - Predicting the effects of nectar robbing on plant reproduction: implications of pollen limitation and plant mating system. AB - The outcome of species interactions is often difficult to predict, depending on the organisms involved and the ecological context. Nectar robbers remove nectar from flowers, often without providing pollination service, and their effects on plant reproduction vary in strength and direction. In two case studies and a meta analysis, we tested the importance of pollen limitation and plant mating system in predicting the impacts of nectar robbing on female plant reproduction. We predicted that nectar robbing would have the strongest effects on species requiring pollinators to set seed and pollen limited for seed production. Our predictions were partially supported. In the first study, natural nectar robbing was associated with lower seed production in Delphinium nuttallianum, a self compatible but non-autogamously selfing, pollen-limited perennial, and experimental nectar robbing reduced seed set relative to unrobbed plants. The second study involved Linaria vulgaris, a self-incompatible perennial that is generally not pollen limited. Natural levels of nectar robbing generally had little effect on estimates of female reproduction in L. vulgaris, while experimental nectar robbing reduced seed set per fruit but not percentage of fruit set. A meta-analysis revealed that nectar robbing had strong negative effects on pollen-limited and self-incompatible plants, as predicted. Our results suggest that pollination biology and plant mating system must be considered to understand and predict the ecological outcome of both mutualistic and antagonistic plant-animal interactions. PMID- 21636389 TI - Widespread mycorrhizal specificity correlates to mycorrhizal function in the neotropical, epiphytic orchid Ionopsis utricularioides (Orchidaceae). AB - Tropical orchids constitute the greater part of orchid diversity, but little is known about their obligate mycorrhizal relationships. The specificity of these interactions and associated fungal distributions could influence orchid distributions and diversity. We investigated the mycorrhizal specificity of the tropical epiphytic orchid Ionopsis utricularioides across an extensive geographical range. DNA ITS sequence variation was surveyed in both plants and mycorrhizal fungi. Phylogeographic relationships were estimated for the mycorrhizal fungi. Orchid functional outcomes were determined through in vitro seed germination and seedling growth with a broad phylogenetic representation of fungi. Most fungal isolates derived from one clade of Ceratobasidium (anamorphs assignable to Ceratorhiza), with 78% within a narrower phylogenetic group, clade B. No correlation was found between the distributions of orchid and fungal genotypes. All fungal isolates significantly enhanced seed germination, while fungi in clade B significantly enhanced seedling growth. These results show that I. utricularioides associates with a phylogenetically narrow, effective fungal clade over a broad distribution. This preference for a widespread mycorrhizae may partly explain the ample distribution and abundance of I. utricularioides and contrasts with local mycorrhizal diversification seen in some nonphotosynthetic orchids. Enhanced orchid function with a particular fungal subclade suggests mycorrhizal specificity can increase orchid fitness. PMID- 21636390 TI - Comparative height crown allometry and mechanical design in 22 tree species of Kuala Belalong rainforest, Brunei, Borneo. AB - In rainforests, trunk size, strength, crown position, and geometry of a tree affect light interception and the likelihood of mechanical failure. Allometric relationships of tree diameter, wood density, and crown architecture vs. height are described for a diverse range of rainforest trees in Brunei, northern Borneo. The understory species follow a geometric model in their diameter-height relationship (slope, beta = 1.08), while the stress-elasticity models prevail (beta = 1.27-1.61) for the midcanopy and canopy/emergent species. These relationships changed with ontogeny, especially for the understory species. Within species, the tree stability safety factor (SSF) and relative crown width decreased exponentially with increasing tree height. These trends failed to emerge in across-species comparisons and were reversed at a common (low) height. Across species, the relative crown depth decreased with maximum potential height and was indistinguishable at a common (low) height. Crown architectural traits influence SSF more than structural property of wood density. These findings emphasize the importance of applying a common reference size in comparative studies and suggest that forest trees (especially the understory group) may adapt to low light by having deeper rather than wider crowns due to an efficient distribution and geometry of their foliage. PMID- 21636391 TI - Genomic relationships between the cultivated peanut (Arachis hypogaea, Leguminosae) and its close relatives revealed by double GISH. AB - Arachis hypogaea is a natural, well-established allotetraploid (AABB) with 2n = 40. However, researchers disagree on the diploid genome donor species and on whether peanut originated by a single or multiple events of polyploidization. Here we provide evidence on the genetic origin of peanut and on the involved wild relatives using double GISH (genomic in situ hybridization). Seven wild diploid species (2n = 20), harboring either the A or B genome, were tested. Of all genomic DNA probe combinations assayed, A. duranensis (A genome) and A. ipaensis (B genome) appeared to be the best candidates for the genome donors because they yielded the most intense and uniform hybridization pattern when tested against the corresponding chromosome subsets of A. hypogaea. A similar GISH pattern was observed for all varieties of the cultigen and also for A. monticola. These results suggest that all presently known subspecies and varieties of A. hypogaea have arisen from a unique allotetraploid plant population, or alternatively, from different allotetraploid populations that originated from the same two diploid species. Furthermore, the bulk of the data demonstrated a close genomic relationship between both tetraploids and strongly supports the hypothesis that A. monticola is the immediate wild antecessor of A. hypogaea. PMID- 21636392 TI - High salinity alters chloroplast morpho-physiology in a freshwater Kirchneriella species (Selenastraceae) from Ethiopian Lake Awasa. AB - Plants differ in their ability to tolerate salt stress. In aquatic ecosystems, it is important to know the responses of microalgae to increased salinity levels, especially considering that global warming will increase salinity levels in some regions of the Earth, e.g., Ethiopia. A green microalga, Kirchneriella sp. (Selenastraceae, Chlorophyta), isolated from freshwater Lake Awasa in the Rift Valley, Ethiopia, was cultured in media amended with 0, 0.4, 1.9, 5.9, and 19.4 g NaCl.L(-1) adjusted with NaCl to five salinity levels adjusted with NaCl. Growth was monitored for 3 mo, then samples were collected for photosynthetic pigment determinations, microspectrofluorimetric analyses, and micro- and submicroscopic examinations. The best growth was found at 1.9 g NaCl.L(-1). In the chloroplast, excess NaCl affected the coupling of light harvesting complex II and photosystem II (LHCII-PSII), but changes in thylakoid architecture and in the PSII assembly state allowed sufficient integrity of the photosynthetic membrane. The mucilaginous capsule around the cell probably provided partial protection against NaCl excess. On the whole, the microalga is able to acclimate to a range of NaCl concentrations, and this plasticity indicates that Kirchneriella sp. may survive future changes in water quality. PMID- 21636393 TI - Maternal effects of drought stress and inbreeding in Impatiens capensis (Balsaminaceae). AB - Maternal effects can have substantial impacts on plant fitness and plant populations. Stressful environmental conditions can cause a maternal plant to inadequately provision its progeny, resulting in poor seedling growth, low reproductive success, and decreased competitive ability. Maternal effects consist of environmental and genetic load components, but the interactions between these two components have rarely been considered. To determine the effects of maternal drought stress and maternal inbreeding on progeny biomass (a fitness correlate) and physiological responses to drought stress, we conducted a greenhouse experiment with genetic lines from two populations (mesic site vs. dry site) of the herbaceous annual Impatiens capensis (Balsaminaceae). Seeds were collected from cleistogamous flowers of inbred or outcrossed maternal plants that were subject to either a drought or control treatment. These seeds were grown into juvenile plants that were also subject to either a drought stress or a control treatment. Plants from the mesic site had significantly reduced biomass from maternal drought stress, while plants from the dry site maintained biomass despite adverse maternal environmental conditions. Juvenile plants of both populations had reduced biomass only as a result of maternal inbreeding. Interestingly, inbreeding depression was more apparent when maternal environmental conditions were benign. PMID- 21636394 TI - The taxonomic distribution of C4 photosynthesis in Amaranthaceae sensu stricto. AB - C(4) photosynthesis evolved multiple times in the Amaranthaceae s.s., but the C(4) evolutionary lineages are unclear because the photosynthetic pathway is unknown for most species of the family. To clarify the distribution of C(4) photosynthesis in the Amaranthaceae, we determined carbon isotope ratios of 607 species and mapped these onto a phylogeny determined from matK/trnK sequences. Approximately 28% of the Amaranthaceae species use the C(4) pathway. C(4) species occur in 10 genera-Aerva, Amaranthus, Blutaparon, Alternanthera, Froelichia, Lithophila, Guilleminea, Gomphrena, Gossypianthus, and Tidestromia. Aerva, Alternanthera, and Gomphrena contain both C(3) and C(4) species. In Aerva, 25% of the sampled species are C(4). In Alternanthera, 19.5% are C(4), while 89% of the Gomphrena species are C(4). Integration of isotope and matK/trnK data indicated C(4) photosynthesis evolved five times in the Amaranthaceae, specifically in Aerva, Alternanthera, Amaranthus, Tidestromia, and a lineage containing Froelichia, Blutaparon, Guilleminea, Gomphrena pro parte, and Lithophila. Aerva and Gomphrena are both polyphyletic with C(3) and C(4) species belonging to distinct clades. Alternanthera appears to be monophyletic with C(4) photosynthesis originating in a terminal sublineage of procumbent herbs. Alpine C(4) species were also identified in Alternanthera, Amaranthus, and Gomphrena, including one species (Gomphrena meyeniana) from 4600 m a.s.l. PMID- 21636395 TI - Phylogenetic systematics and character evolution in the angiosperm family Haloragaceae. AB - The poorly known Haloragaceae R. Br. (Saxifragales) are highly diverse in habit (small trees to submerged aquatics) and labile in floral merosity (2-4), both uncommon among the core eudicots. This family has a cosmopolitan distribution, but taxonomic diversity is concentrated in Australia. An explicit phylogenetic approach has not previously been utilized to examine relationships or character evolution in this family. We used molecular evidence from nrDNA ITS and cpDNA trnK and matK regions under both Bayesian and parsimony analyses to address phylogenetic relationships. Combined molecular analyses defined a monophyletic Haloragaceae with the woody genera (Haloragodendron, Glischrocaryon) sister to the rest. Relationships among many genera were well resolved, with genera as currently delimited generally well supported, although there were notable exceptions; a new genus (Trihaloragis) is recognized, and the aquatic genus Meionectes is again distinct from Haloragis. Three new species combinations are also recognized. There are multiple (two or three) origins of the submerged aquatic habit in the family and potentially an intermediate reversal to the terrestrial habit, neither previously demonstrated in a core eudicot family using an explicit phylogenetic hypothesis. Ancestral character analyses suggest two origins of trimerous flowers and multiple reductions to dimerous flowers throughout Haloragaceae. PMID- 21636396 TI - Molecular phylogenetics of tribe Poranthereae (Phyllanthaceae; Euphorbiaceae sensu lato). AB - Novel insights into the evolutionary history of a taxonomically complex tropical plant group were gained in this study using DNA sequence data. A molecular phylogenetic analysis of the newly circumscribed and expanded tribe Poranthereae (Phyllanthaceae) is presented. Sampling included 97 accessions for 63 of c. 120 species. Largely congruent results have been obtained from nuclear ribosomal ITS and plastid matK sequences. These analyses support the recognition of Andrachne and Leptopus as distinct genera. The deceptively similar Andrachne section Phyllanthopsis, Andrachne ovalis, and Leptopus decaisnei are separate lineages to be segregated. Zimmermannia and Zimmermanniopsis are embedded in Meineckia; Oreoporanthera is embedded in Poranthera; and Archileptopus is embedded in Leptopus. Andrachne section Pseudophyllanthus is polyphyletic, the two Madagascan endemics emerging as a sister clade to Meineckia. The noncontiguous distributions of Andrachne sensu lato and Leptopus sensu lato were found to be the result of separate evolutionary histories of morphologically similar clades, whereas Andrachne sensu stricto and Meineckia remain geographically disjunct. Actephila and Leptopus are sisters with a sympatric distribution in humid Asia. Presence of petals appears to be plesiomorphic for the tribe. Petals are reduced in Actephila and Oreoporanthera and lost in the Meineckia clade. PMID- 21636397 TI - Potomacanthus lobatus gen. et sp. nov., a new flower of probable Lauraceae from the Early Cretaceous (Early to Middle Albian) of eastern North America. AB - A charcoalified fossil flower, Potomacanthus lobatus gen. et sp. nov., is described from the Early Cretaceous (Early to Middle Albian) Puddledock locality, Virginia, USA. Internal floral structure was studied using nondestructive synchrotron-radiation x-ray tomographic microscopy (SRXTM). The flower is bisexual and trimerous. The perianth consists of two whorls of tepals. The androecium has two whorls of fertile stamens. Anthers open by two distally hinged valves. The gynoecium consists of a single carpel that is plicate in the style and ascidiate in the ovary and contains a single pendant ovule. The fossil flower shares many similarities with flowers of extant Lauraceae and is unlike flowers of other families of Laurales. However, the fossil flower also differs in detail from all extant or fossil Lauraceae, particularly in configuration of the androecium. The new taxon, together with previously described but more fragmentary material from the Puddledock locality, provides the earliest fossil record of plants more closely related to Lauraceae than to any other extant family. It reveals several derived morphological characters that are potential synapomorphies among extant representatives of the family Lauraceae and contributes to the growing evidence for an early diversification of Laurales before the end of the Early Cretaceous. PMID- 21636398 TI - Quantifying the presence and absence of turgor for the spatial characterization of cortical senescence in roots of Triticum aestivum (Poaceae). AB - Cortical senescence is an important feature of the roots of a number of graminaceous species because it may contribute to rhizodeposition of carbon and influence the efficiency of nutrient uptake. A major limitation to understanding the physiological control of senescence and its impact on rhizosphere processes has been the lack of reliable techniques for characterizing the progress of senescence along the root. The use of a single-cell pressure probe was evaluated for quantifying cell integrity in different regions of wheat roots. The percentage of locations with turgid cells declined with increasing distance behind the root apex. The decline preceded visible collapse of the cortex but after the loss of nuclear staining in the outer cortex. The percentage of locations with turgid cells was closely associated with root diameter, which suggests that measurements of changes in diameter, made using minirhizotrons under well-watered conditions in the field, could provide estimates of in situ rates of cortical senescence. PMID- 21636399 TI - Phylogenetic evidence for a flower size and number trade-off. AB - The size and number of flowers displayed together on an inflorescence (floral display) influences pollinator attraction and pollen transfer and receipt, and is integral to plant reproductive success and fitness. Life history theory predicts that the evolution of floral display is constrained by trade-offs between the size and number of flowers and inflorescences. Indeed, a trade-off between flower size and flower number is a key assumption of models of inflorescence architecture and the evolution of floral display. Surprisingly, however, empirical evidence for the trade-off is limited. In particular, there is a lack of phylogenetic evidence for a trade-off between flower size and number. Analyses of phylogenetic independent contrasts (PICs) of 251 angiosperm species spanning 63 families yielded a significant negative correlation between flower size and flower number. At smaller phylogenetic scales, analyses of individual genera did not always find evidence of a trade-off, a result consistent with previous studies that have examined the trade-off for a single species or genus. Ours is the first study to support an angiosperm-wide trade-off between flower size and number and supports the theory that life history constraints have influenced the evolution of floral display. PMID- 21636400 TI - Analysis of substage associations in prophase I of meiosis in floral buds of wild type Arabidopsis thaliana (Brassicaceae). AB - We developed an improved cytological protocol for producing high quality, light microscope images of plant meiotic chromosomes. Because the technique works on species with small genomes and thick microsporocyte cell walls, it should be useful for studying the wild relatives of Arabidopsis and other eudicots with small genomes. Combining this improved fixation protocol with our new analysis of associated substages in floral buds, we can unambiguously assign individual meiotic cells to particular substages of prophase I in Arabidopsis thaliana, even for difficult distinctions such as that between late zygotene or early diplotene. In this report we provide the first estimate of the individual duration of the zygotene and pachytene substages (4.8 h and 10.0 h, respectively) in A. thaliana. We also have examined the diffuse substage of prophase I and report that during this post-pachytene substage, nuclei retain the association of homologous nucleolus organizer regions and homologous centromeres, despite the generally diffuse chromatin and generally unpaired chromosome regions. Additionally, we have observed that centromeric regions of the chromosomes of diffuse-stage nuclei are highly condensed, more so than those of any other substage of prophase I. PMID- 21636401 TI - Comparison of whole chloroplast genome sequences to choose noncoding regions for phylogenetic studies in angiosperms: the tortoise and the hare III. AB - Although the chloroplast genome contains many noncoding regions, relatively few have been exploited for interspecific phylogenetic and intraspecific phylogeographic studies. In our recent evaluation of the phylogenetic utility of 21 noncoding chloroplast regions, we found the most widely used noncoding regions are among the least variable, but the more variable regions have rarely been employed. That study led us to conclude that there may be unexplored regions of the chloroplast genome that have even higher relative levels of variability. To explore the potential variability of previously unexplored regions, we compared three pairs of single-copy chloroplast genome sequences in three disparate angiosperm lineages: Atropa vs. Nicotiana (asterids); Lotus vs. Medicago (rosids); and Saccharum vs. Oryza (monocots). These three separate sequence alignments highlighted 13 mutational hotspots that may be more variable than the best regions of our former study. These 13 regions were then selected for a more detailed analysis. Here we show that nine of these newly explored regions (rpl32 trnL((UAG)), trnQ((UUG))-5'rps16, 3'trnV((UAC))-ndhC, ndhF-rpl32, psbD trnT((GGU)), psbJ-petA, 3'rps16-5'trnK((UUU)), atpI-atpH, and petL-psbE) offer levels of variation better than the best regions identified in our earlier study and are therefore likely to be the best choices for molecular studies at low taxonomic levels. PMID- 21636402 TI - Evolution of Lecythidaceae with an emphasis on the circumscription of neotropical genera: information from combined ndhF and trnL-F sequence data. AB - The Lecythidaceae comprise a pantropical family best known for the edible seeds of the Brazil nut (Bertholletia excelsa) and the cannon-ball tree (Couroupita guianensis), which is planted as a botanical curiosity in subtropical and tropical gardens. In addition, species of the family are often among the most common in neotropical forests, especially in the Amazon Basin. The Brazil nut family is diverse and abundant in the Amazon and is considered to be an indicator of undisturbed or scarcely disturbed lowland forests; thus, what is learned about its evolution, ecology, and biogeography may suggest similar patterns for other Amazonian tree families. We used combined data sets derived from the ndhF and trnL-F genes to elucidate relationships of genera in both the Old and New Worlds that have been associated with Lecythidaceae. Our molecular tree agrees with the recognition of Napoleonaeaceae and Scytopetalaceae. Within the Lecythidaceae, there is molecular support for recognizing three subfamilies: Foetidioideae, Planchonioideae, and Lecythidoideae. We then focused on genera of the Lecythidoideae and found support for recognizing Allantoma (when the actinomorphic-flowered species of Cariniana are included in it), Grias, Gustavia, Corythophora, Couratari, and Couroupita, but conclude that Cariniana, Lecythis, and Eschweilera are not monoyphyletic. Because the position of the monotypic Bertholletia excelsa in relation to the other zygomorphic-flowered genera is not resolved, we are not able to comment on its generic relationships. PMID- 21636403 TI - A comparative analysis of the Lactuca and Helianthus (Asteraceae) plastid genomes: identification of divergent regions and categorization of shared repeats. AB - We have sequenced two complete chloroplast genomes in the Asteraceae, Helianthus annuus (sunflower), and Lactuca sativa (lettuce), which belong to the distantly related subfamilies, Asteroideae and Cichorioideae, respectively. The Helianthus chloroplast genome is 151 104 bp and the Lactuca genome is 152 772 bp long, which is within the usual size range for chloroplast genomes in flowering plants. When compared to tobacco, both genomes have two inversions: a large 22.8-kb inversion and a smaller 3.3-kb inversion nested within it. Pairwise sequence divergence across all genes, introns, and spacers in Helianthus and Lactuca has resulted in the discovery of new, fast-evolving DNA sequences for use in species-level phylogenetics, such as the trnY-rpoB, trnL-rpl32, and ndhC-trnV spacers. Analysis and categorization of shared repeats resulted in seven classes useful for future repeat studies: double tandem repeats, three or more tandem repeats, direct repeats dispersed in the genome, repeats found in reverse complement orientation, hairpin loops, runs of A's or T's in excess of 12 bp, and gene or tRNA similarity. Results from BLAST searches of our genomic sequence against expressed sequence tag (EST) databases for both genomes produced eight likely RNA edited sites (C -> U changes). These detailed analyses in Asteraceae contribute to a broader understanding of plastid evolution across flowering plants. PMID- 21636404 TI - Searching for the relatives of Coffea (Rubiaceae, Ixoroideae): the circumscription and phylogeny of Coffeeae based on plastid sequence data and morphology. AB - The circumscription of Coffeeae (Rubiaceae) and phylogenetic relationships within the tribe were evaluated using sequence data from four plastid regions (trnL-F intron, trnL-F intergenic spacer [IGS], rpl16 intron, and accD-psa1 IGS) and a morphological data set. Eleven candidates for inclusion in Coffeeae were examined using plastid data, and a further three were investigated using morphology alone. Based on previous phylogenetic analysis of the subfamily Ixoroideae, nine genera representing five tribes were used as outgroups. Our results support an enlarged circumscription for Coffeeae, containing 11 genera, viz. Argocoffeopsis, Belonophora, Calycosiphonia, Coffea, Diplospora, Discospermum, Nostolachma, Psilanthus, Tricalysia, Sericanthe, and Xantonnea. The inclusion of Diplospora and Tricalysia within Coffeeae, based on published molecular data, and the inclusion of Argocoffeopsis, Belonophora, Calycosiphonia, Discospermum, and Sericanthe, based on morphological evidence, are well supported. Nostolachma is newly transferred from Gardenieae subtribe Diplosporinae to Coffeeae, and Xantonnea from Octotropideae to Coffeeae. The exclusion of Bertiera from Coffeeae and placement in tribe Bertiereae is supported on the basis of molecular and morphological data. The removal of Diplospora and all other genera from Gardenieae subtribe Diplosporinae to Coffeeae and Octotropideae renders Diplosporinae superfluous. It is proposed that Xantonneopsis be transferred to Octotropideae; Petitiocodon is tentatively placed in Gardenieae. The monophyly of seven genera is supported, but Coffea is identified as paraphyletic in relation to Psilanthus on the basis of molecular and combined molecular and morphological data. PMID- 21636405 TI - Origin and evolution of North American polyploid Silene (Caryophyllaceae). AB - Nuclear DNA sequences from introns of the low-copy nuclear gene family encoding the second largest subunit of RNA polymerases and the ribosomal internal transcribed spacer (ITS) regions, combined with the psbE-petL spacer and the rps16 intron from the chloroplast genome were used to infer origins and phylogenetic relationships of North American polyploid Silene species and their closest relatives. Although the vast majority of North American Silene species are polyploid, which contrasts to the diploid condition dominating in other parts of the world, the phylogenetic analyses rejected a single origin of the North American polyploids. One lineage consists of tetraploid Silene menziesii and its diploid allies. A second lineage, Physolychnis s.l., consists of Arctic, European, Asian, and South American taxa in addition to the majority of the North American polyploids. The hexaploid S. hookeri is derived from an allopolyploidization between these two lineages. The tetraploid S. nivea does not belong to any of these lineages, but is closely related to the European diploid S. baccifera. The poor resolution within Physolychnis s.l. may be attributed to rapid radiation, recombination among homoeologues, homoplasy, or any combination of these factors. No extant diploid donors could be identified in Physolychnis s.l. PMID- 21636406 TI - Cascadiacarpa spinosa gen. et sp. nov. (Fagaceae): castaneoid fruits from the Eocene of Vancouver Island, Canada. AB - Documenting the paleodiversity of well-studied angiosperm families serves to broaden their circumscription while also providing a time-specific reference point to mark the first occurrence of characters and appearance of lineages. More than 80 anatomically preserved specimens of spiny, cupulate fruits in various developmental stages have been studied from the Eocene Appian Way locality of Vancouver Island, British Columbia, Canada. Details of internal anatomy and external morphology are known for the cupules, fruits, and pedicels. Cupule spines branch and are often borne in clusters. Cupules lack clear sutures and are adnate to a single nut that is enclosed entirely with the exception of the apical stylar protrusion of the pistil. A central hollow cylinder of vascular tissue can be seen extending up the peduncle to the base of the fruit and along the inner wall of the cupule. The fruit has a sclerotic outer pericarp that grades into a parenchymatous mesocarp and a sclerotic endocarp lining the locules. Early in development, the two locules are divided by a thin septum to which the ovules are attached. Only one seed develops to maturity as evidenced by an embryo occupying the locule alongside an abortive apical ovule. Three-dimensional reconstructions of these fruits have allowed for comparisons to both extinct and extant fagaceous taxa. The Appian Way fruits are most similar to extant Castanopsis species (Fagaceae) but differ in having only two locules. Cascadiacarpa spinosa gen. et sp. nov. Mindell, Stockey et Beard is the first occurence of a bipartite gynoecium and earliest known occurrence of hypogeous fruits in Fagaceae. The appearance of Casacadiacarpa in the Eocene of British Columbia supports a Paleogene radiation of the family. The numerous derived characters of these fruits show that evalvate, spiny, single-fruited cupules of Fagaceae were present in the Paleogene of North America. PMID- 21636407 TI - Diversity of Kranz anatomy and biochemistry in C4 eudicots. AB - C(4) photosynthesis and Kranz anatomy occur in 16 eudicot families, a striking example of convergent evolution. Biochemical subtyping for 13 previously undiagnosed C(4) eudicot species indicated that 10 were NADP-malic enzyme (ME) and three were NAD-ME. A total of 33 C(4) species, encompassing four Kranz anatomical types (atriplicoid, kochioid, salsoloid, and suaedioid), and 21 closely related C(3) species were included in a quantitative anatomical study in which we found that, unlike similar studies in grasses and sedges, anatomical type had no predictive value for the biochemical subtype. In a multivariate canonical discriminant analysis, C(4) species were distinguished from C(3) species by the mesophyll to bundle sheath ratio and exposure of the bundle sheath surface to intercellular space. Discrimination between NADP-ME and NAD-ME was not significant, although in a Mantel test grouping by biochemical subtype was significant, while grouping by family was not. This comprehensive survey of C(4) anatomy and biochemistry unequivocally demonstrated that atriplicoid anatomy and NADP-ME biochemistry predominate in many evolutionary lineages. In addition to a main decarboxylating enzyme, high activity of a second decarboxylating enzyme was often observed. Notably, PEP-carboxykinase activity was significant in a number of species, demonstrating that this enzyme could also serve as a secondary pathway for C(4) metabolism in eudicots. PMID- 21636408 TI - Key innovations in the evolution of Kranz anatomy and C4 vein pattern in Flaveria (Asteraceae). AB - Kranz anatomy and C(4) vein pattern are required for C(4) biochemical functioning in C(4) plants; however, the evolutionary timing of anatomical and biochemical adaptations is unknown. From the genus Flaveria, 16 species (C(3), C(4), intermediates [C(3)-C(4), C(4)-like]) were analyzed, novel anatomical and vein pattern characters were analyzed and key anatomical differences among photosynthetic groups were highlighted. A stepwise acquisition of anatomical and vein pattern traits prior to derived biochemistry was outlined on the basis of the phylogeny of Flaveria. Increased vein density represents a potential "precondition" contributing to lower ratios of photosynthetic tissues (mesophyll, bundle sheath) and precedes further anatomical and biochemical modifications observed in derived C(3)-C(4) intermediates. In derived Flaveria species, bundle sheath volume is modified through cell expansion, whereas mesophyll volume is altered through mesophyll cell expansion, reductions in the number of ground tissue layers, and increased vein density. Results demonstrated that key anatomical features of C(4) plants are also required for C(3)-C(4) biochemical intermediacy, and anatomical and biochemical alterations acquired during evolution of intermediacy may predispose a species for evolution of C(4) photosynthesis. C(4)-like species are similar to C(4) species, demonstrating that Kranz anatomy is fully evolved before complete C(4) biochemistry is achieved. PMID- 21636409 TI - Molecular phylogenetic evidence for the origin of a diploid hybrid of Paeonia (Paeoniaceae). AB - There is growing evidence that hybridization not only by means of allopolyploidy but also at the homoploidy level was a major driving force of plant diversification. While allopolyploidy is known to be a common mode of speciation in Paeonia (Paeoniaceae), hybrid speciation at the diploid level needs further evaluation. Paeonia anomala was previously considered to be an interspecific hybrid but with an unknown ploidy level. In this study P. anomala is identified as a diploid (2n = 10). With increased sampling of populations and molecular markers, we showed that P. anomala is a homoploid hybrid that originated from a cross between P. veitchii and P. lactiflora. Five populations of P. anomala were sequenced for the following molecular markers: the matK gene and two intergenic spacers, psbA-trnH and rps16-trnQ, of the chloroplast genome; the internal transcribed spacers (ITS) of nuclear ribosomal DNA; and three low-copy nuclear genes, Adh1, Adh2, and Gpat. The populations of P. anomala were grouped together with P. veitchii on the ITS and Gpat phylogenies but with P. lactiflora on the chloroplast phylogeny. Sequence polymorphism was found at the Adh1 and Adh2 loci within individuals of P. anomala. These polymorphic sequences were grouped with P. veitchii and P. lactiflora, respectively. Phenetic analysis indicated that P. anomala is morphologically similar to P. veitchii. Phenotypic evolution resulting from the combination of two diverged genomes might have occurred primarily at the physiological level and allowed P. anomala to adapt to geographic regions different from those of its parents. PMID- 21636410 TI - Do differences in plant and flower age change mating patterns and alter offspring fitness in Raphanus sativus (Brassicaceae)? AB - When more pollen is present on stigmas than needed to fertilize all ovules, selection among pollen grains may occur due to effects of both pollen donors and maternal plants. We asked whether increasing plant age and flower age, two changes in maternal condition, altered the pattern of seed paternity after mixed pollination. We also asked whether changes in seed paternity affected offspring success in an experimental garden. While flower age did not affect seed paternity, there was a dramatic shift in pollen donor performance as plants aged. These differences were seen in the offspring as well, where the offspring of one pollen donor, which sired more seeds on young plants, flowered earlier in the season, and the offspring of another pollen donor, which sired more seeds on old plants, flowered later in the season. Thus, change in maternal condition resulted in altered seed paternity, perhaps because the environment for pollen tube growth was different. The pattern of seed paternity and offspring performance suggests that pollen donors may show temporal specialization. PMID- 21636411 TI - Relative pollination effectiveness of floral visitors of Pitcairnia angustifolia (Bromeliaceae). AB - The effectiveness of flower visitors as pollinators will determine their potential role as selective agents on flower traits. Pitcairnia angustifolia has floral characters that would fit pollination by long-billed hummingbirds, and they should be the most effective pollinators for this plant. To test this prediction, we characterized the behavior of visitors toward flowers and their pollination effectiveness. Coereba flaveola (bananaquits) was the most frequent flower visitor and acted as a primary nectar robber; however, they pollinated incidentally and deposited pollen on stigmas. The endemic short-billed hummingbird Chlorostilbon maugaeus behaved as a secondary robber and did not pollinate flowers. As expected, the long-billed hummingbird, Anthracothorax viridis, was the most efficient visitor in terms of pollen deposition; however, it was the least frequent flower visitor. Introduced Apis mellifera (honeybees) were second in efficiency at depositing pollen and performed one third of the flower visits. Estimates of the expected rate of pollen deposition by each pollinator did not identify a single most effective pollinator. For P. angustifolia at least three flower visitors including an exotic bee and a nectar robber may be equally important to reproductive success. While these results limit our ability to make predictions on the role of hummingbird-pollination on current flower evolution, they do suggest the potential for pollination redundancy among flower visitors for P. angustifolia populations. PMID- 21636412 TI - Genotype, soil type, and locale effects on reciprocal transplant vigor, endophyte growth, and microbial functional diversity of a narrow sagebrush hybrid zone in Salt Creek Canyon, Utah. AB - When addressing the nature of ecological adaptation and environmental factors limiting population ranges and contributing to speciation, it is important to consider not only the plant's genotype and its response to the environment, but also any close interactions that it has with other organisms, specifically, symbiotic microorganisms. To investigate this, soils and seedlings were reciprocally transplanted into common gardens of the big sagebrush hybrid zone in Salt Creek Canyon, Utah, to determine location and edaphic effects on the fitness of parental and hybrid plants. Endophytic symbionts and functional microbial diversity of indigenous and transplanted soils and sagebrush plants were also examined. Strong selection occurred against the parental genotypes in the middle hybrid zone garden in middle hybrid zone soil; F(1) hybrids had the highest fitness under these conditions. Neither of the parental genotypes had superior fitness in their indigenous soils and habitats; rather F(1) hybrids with the nonindigenous maternal parent were superiorly fit. Significant garden-by-soil type interactions indicate adaptation of both plant and soil microorganisms to their indigenous soils and habitats, most notably in the middle hybrid zone garden in middle hybrid zone soil. Contrasting performances of F(1) hybrids suggest asymmetrical gene flow with mountain, rather than basin, big sagebrush acting as the maternal parent. We showed that the microbial community impacted the performance of parental and hybrid plants in different soils, likely limiting the ranges of the different genotypes. PMID- 21636413 TI - Phenotypic integration and the potential for independent color evolution in a polymorphic spring ephemeral. AB - The genetic architecture of the total phenotype may substantially constrain or enhance the evolution of floral color within populations in response to multiple selection pressures. Using Claytonia virginica I previously identified opposing selection on floral color generated through herbivores and pathogens. Here I ask whether the evolution of floral color in this system is constrained or unconstrained by its phenotypic integration with floral and vegetative traits. Morphological, physiological, and pollen traits were measured on over 400 plants in the field and greenhouse, and these data were used to test whether floral color morphs differed with respect to other traits and whether the among-trait correlation structure differed across the color morphs. The color morphs varied with respect to most measured traits; however, the pattern of variation was not consistent among them, and there was little evidence of trade-offs with floral color. A common principal components analysis revealed that the pattern of phenotypic integration substantially differed among the color morphs. Combined, these results suggest that floral-color evolution may proceed relatively unconstrained by associations with other traits in this system. The absence of a strong constraint in combination with known fluctuating selective pressures may help to explain observed within- and among-population color variation in this species. PMID- 21636414 TI - Leaf damage and gender but not flower damage affect female fitness in Nemophila menziesii (Hydrophyllaceae). AB - Researchers can answer questions about the evolution or maintenance of separate sexes using dioecious plant systems. Because females in these species typically put more resources into reproductive effort than male plants, researchers have hypothesized that females may be less tolerant of the stresses found in marginal habitats. Herbivory can act as a biotic stressor that reduces resources in plants much like a marginal habitat can. Females may be limited by resources, and may thus be less tolerant to herbivory than males. Here, I explore the relationships between florivory, leaf herbivory, and gender in a gynodioecious, annual plant, Nemophila menziesii (Hydrophyllaceae, senso lato). I performed a crossed design experiment examining the main effects and interactions of plant gender, artificial leaf damage, and artificial flower damage on components of female plant fitness. Leaf damage decreased fruit set and females made significantly more fruit than hermaphrodites. However, contrary to theory, I found little evidence for a gender by damage interaction for either type of artificial herbivory. Based on these results, I propose more work exploring the effects of both source and sink damage in dioecious species to help elucidate where and when different sexual morphs are favored by natural selection. PMID- 21636415 TI - Ecological and evolutionary determinants of a key plant functional trait: wood density and its community-wide variation across latitude and elevation. AB - Wood density is an important trait in trees indicative of their life history and mechanical and physiological strategies. The following examines the evolutionary ecology of wood density using a large database for seed plants. In particular, we focused on the geographic and phylogenetic variation in wood density for both gymnosperms and angiosperms. A phylogenetic supertree was constructed for over 4600 taxa, allowing for comprehensive analyses of divergences across the seed plant phylogeny. Community-wide means and variances for wood densities were quantified for 171 standardized forest communities. Wood density was generally highly conserved across the entire seed plant phylogeny, yet large divergences were found within the rosid clade. Geographic and community variation in wood density, however, was significantly lower in temperate and high elevation communities, dominated by gymnosperms, than in tropical lowland communities, dominated by angiosperms, suggesting an increase in trait and, to some extent, clade filtering with latitude and elevation. Together, our results support the notion that both biotic and abiotic forces have been important in the evolution of wood density as well as in controlling the observed trait mean and variance across geographic gradients. PMID- 21636416 TI - Development of the enigmatic peristome of Timmia megapolitana (Timmiaceae; Bryophyta). AB - The Timmiaceae (Bryophyta) have been traditionally classified within the Bryales based on peristome architecture. Phylogenetic analyses of nucleotide sequences have revealed relationships that are incongruent with this hypothesis and have implicated an origin for this lineage early in the radiation of arthrodontous mosses (Bryopsida). This unexpected phylogenetic placement raises important questions about the evolutionary significance of the Timmia peristome, which differs from all other mosses by 64 isomorphic filaments topping the endostomial membrane. A developmental study of the peristome in Timmia megapolitana was undertaken to examine alignments of anticlinal cell walls in the inner peristomial layer (IPL) with those of the primary peristomial layer (PPL), a character that has been used to define major arthrodontous lineages. Criteria were established for assessing longitudinally homologous regions that contribute to the peristome-forming region. Young sporophytes were examined using histological techniques, and the alignment of the cell wall divisions was quantified. Critical divisions in the IPL of T. megapolitana were determined to be symmetrical, similar to patterns in the Funariales. This research provides novel developmental observations for a putative ancestral lineage of arthrodontous mosses, reevaluates criteria used to compare developmental studies of different lineages and discusses the phylogenetic implications of these observations. PMID- 21636417 TI - Mechanisms of prezygotic reproductive isolation between two sympatric species, Gelsemium rankinii and G. sempervirens (Gelsemiaceae), in the southeastern United States. AB - Natural hybridization plays a critical role in speciation, the maintenance of reproductive isolation, and genetic introgression. While many plant species have hybrid swarms in areas of sympatry, the lack of hybrids among closely related sympatrically distributed species suggests that strong pre- and/or postzygotic barriers exist to hybridization. Gelsemium sempervirens and G. rankinii (Gelsemiaceae) are sympatrically distributed southeastern sister taxa that have strong postzygotic barriers to hybrid formation and high levels of genetic differentiation. In this study, two sympatric populations in Lowndes County, Georgia were surveyed from 1999-2005 to assess the role of temporal and pollinator isolation as potential prezygotic barriers. The populations had mostly non-overlapping flowering periods in 2003-2005, with significant differences in time of peak flowering and length of flowering. Both species shared a similar community of flower visitors, with the apid bee Habropoda laboriosa the dominant visitor to both species. A choice experiment found that H. laboriosa visited both species but preferred G. sempervirens. The primary prezygotic barrier is temporal isolation preventing hybridization in spite of the shared pollinators. This study suggests that reliance on a shared pollinator during speciation may limit opportunity for divergent selection on flowering time. PMID- 21636418 TI - A molecular approach to species identification of Chenopodiaceae pollen grains in surface soil. AB - Pollen identification and classification are important not only for palynologists, but also for systematists and ecologists. Because palynological methods for the identification of pollen in surface soil until now could resolve at best to the generic level, we have developed a molecular approach to species level identification of Chenopodiaceae pollen in surface soils. Surface soil samples were collected in the central area of Junggar Desert Basin, Xinjiang, China. Fresh leaves of 19 Chenopodiaceae species were sampled for DNA sequencing, establishing a database of internal transcribed spacer (ITS) regions of nuclear ribosomal DNA for Chenopodiaceae. Individual chenopod pollen grains in a soil sample were separated from the soil and the ITS1 region of each pollen grain was amplified using nested PCR and sequenced. By comparing the amplified ITS1 sequences to those in the Chenopodiaceous database, we identified the pollen in the soil samples to the level of species. The new method provides a technical reference for species identification of soil surface pollen for other families. This work is necessary for further efforts to interpret the relationship of surface soil pollen to vegetation characteristics. It also has significant potential for enhancing the ability to identify pollen in clinical airborne allergen or criminological studies. PMID- 21636419 TI - A search for phylogenetically informative wood characters within Lecythidaceae s.l. AB - The wood structure of 71 species representing 24 genera of the pantropical Lecythidaceae s.l., including the edible Brazil nuts (Bertholletia excelsa) and the spectacular cannon-ball tree (Couroupita guianensis), was investigated using light and scanning electron microscopy. This study focused on finding phylogenetically informative characters to help elucidate any obscure evolutionary patterns within the family. The earliest diverging subfamily Napoleonaeoideae has mixed simple/scalariform vessel perforations, scalariform vessel-ray pitting, and high multiseriate rays, all features that are also present in Scytopetaloideae. The wood structure of Napoleonaea is distinct, but its supposed close relative Crateranthus strongly resembles Scytopetaloideae. The isolated position of Foetidia (Foetidioideae) can be supported by a unique type of vessel-ray pitting that is similar in shape and size to intervessel pitting (distinctly bordered, <5 MUm). The more derived Planchonioideae and Lecythidoideae share exclusively simple perforations and two types of vessel-ray pitting, but they can easily be distinguished from each other by the size of intervessel pitting, shape of body ray cells in multiseriate rays, and the type of crystalliferous axial parenchyma cells. The anatomical diversity observed is clearly correlated with differences in plant size (shrubs vs. tall trees): the percentage of scalariform perforations, as well as vessel density, and the length of vessel elements, fibers, and multiseriate rays are negatively correlated with increasing plant size, while the reverse is true for vessel diameter. PMID- 21636420 TI - Pit membranes in tracheary elements of Rosaceae and related families: new records of tori and pseudotori. AB - The micromorphology of pits in tracheary elements was examined in 35 species representing 29 genera of Rosaceae and related families to evaluate the assumption that angiosperm pits are largely invariant. In most Rosaceae, pit membranes between fibers and tracheids frequently appear to have amorphous thickenings with an irregular distribution. Although these structures are torus like under the light microscope, observations by electron microscopy illustrate that they represent "pseudotori" or plasmodesmata-associated thickenings. These thickenings frequently extend from the periphery of the pit membrane and form a cap-like, hollow structure. Pseudotori are occasionally found in few Elaeagnaceae and Rhamnaceae and appear to be related to species with fiber-tracheids and/or tracheids. True tori are strongly associated with round to oval pit apertures and are consistently present in narrow tracheary elements of Cercocarpus (Rosaceae), Planera (Ulmaceae), and ring-porous species of Ulmus and Zelkova (Ulmaceae). Vestured pits with homogenous pit membranes are reported for Hemiptelea (Ulmaceae). The homoplastic nature of pit membrane characteristics may be related to functional adaptations in terms of safety and efficiency of water transport or may reflect different developmental processes of xylem elements. These observations illustrate that there is more variation in angiosperm pits than previously thought. PMID- 21636421 TI - Light-dependent leaf trait variation in 43 tropical dry forest tree species. AB - Our understanding of leaf acclimation in relation to irradiance of fully grown or juvenile trees is mainly based on research involving tropical wet forest species. We studied sun-shade plasticity of 24 leaf traits of 43 tree species in a Bolivian dry deciduous forest. Sampling was confined to small trees. For each species, leaves were taken from five of the most and five of the least illuminated crowns. Trees were selected based on the percentage of the hemisphere uncovered by other crowns. We examined leaf trait variation and the relation between trait plasticity and light demand, maximum adult stature, and ontogenetic changes in crown exposure of the species. Leaf trait variation was mainly related to differences among species and to a minor extent to differences in light availability. Traits related to the palisade layer, thickness of the outer cell wall, and N(area) and P(area) had the greatest plasticity, suggesting their importance for leaf function in different light environments. Short-lived pioneers had the highest trait plasticity. Overall plasticity was modest and rarely associated with juvenile light requirements, adult stature, or ontogenetic changes in crown exposure. Dry forest tree species had a lower light-related plasticity than wet forest species, probably because wet forests cast deeper shade. In dry forests light availability may be less limiting, and low water availability may constrain leaf trait plasticity in response to irradiance. PMID- 21636422 TI - High seedling relative growth rate and specific leaf area are traits of invasive species: phylogenetically independent contrasts of woody angiosperms. AB - Understanding causal factors of exotic species invasions is important not only for prevention and prioritizing control efforts, but also for providing valuable insights into the underlying biology of contrasting life-history strategies. In seedling growth analyses, invasive woody species were compared with less-invasive woody species commonly cultivated in California using phylogenetically corrected procedures (12 phylogenetically independent contrasts). Invasive species were hypothesized to have higher seedling relative growth rates (RGRs) and specific leaf areas (SLAs) than did related less-invasive species. In phylogenetically independent contrasts conducted among taxa within families, high seedling RGRs and SLAs have significant positive associations with woody plant invasiveness. For contrasts containing species invasive in mediterranean regions, invasive species had significantly larger root biomass allocation than did less-invasive species. Optimization of fast seedling growth (high RGR) associated with opportunistic resource acquisition (high SLA) and increased root allocation to survive summer drought may be critical for the success of plant invaders in regions with mediterranean climates. PMID- 21636423 TI - Phenotypic plasticity, precipitation, and invasiveness in the fire-promoting grass Pennisetum setaceum (Poaceae). AB - Invasiveness may result from genetic variation and adaptation or phenotypic plasticity, and genetic variation in fitness traits may be especially critical. Pennisetum setaceum (fountain grass, Poaceae) is highly invasive in Hawaii (HI), moderately invasive in Arizona (AZ), and less invasive in southern California (CA). In common garden experiments, we examined the relative importance of quantitative trait variation, precipitation, and phenotypic plasticity in invasiveness. In two very different environments, plants showed no differences by state of origin (HI, CA, AZ) in aboveground biomass, seeds/flower, and total seed number. Plants from different states were also similar within watering treatment. Plants with supplemental watering, relative to unwatered plants, had greater biomass, specific leaf area (SLA), and total seed number, but did not differ in seeds/flower. Progeny grown from seeds produced under different watering treatments showed no maternal effects in seed mass, germination, biomass or SLA. High phenotypic plasticity, rather than local adaptation is likely responsible for variation in invasiveness. Global change models indicate that temperature and precipitation patterns over the next several decades will change, although the direction of change is uncertain. Drier summers in southern California may retard further invasion, while wetter summers may favor the spread of fountain grass. PMID- 21636424 TI - Aerenchyma development and elevated alcohol dehydrogenase activity as alternative responses to hypoxic soils in the Piriqueta caroliniana complex. AB - The ability of plants to make morphological or physiological adjustments in response to environmental cues allows them to survive and reproduce under a wide range of conditions. One stress that plants are often exposed to is soil oxygen depletion due to flooding. Plants can respond to hypoxic soils by producing oxygen-conducting aerenchymous tissue or through induction of enzymes in the ethanolic fermentation pathway. Here we use greenhouse experiments to examine flood responses in plants of the Piriqueta caroliniana (Turneraceae) complex, which occupy a range of moisture regimes. Morphotypes and hybrids in this complex exhibited contrasting responses to hypoxic conditions. Genotypes from flooded habitats developed aerenchyma and did not substantially elevate levels of alcohol dehydrogenase (ADH) activity, an enzyme associated with anaerobic respiration. Plants from drier sites, on the other hand, did not develop aerenchyma but had much higher levels of ADH activity. Plants with aerenchymous tissue had substantially higher rates of growth under sustained flooding. Results are consistent with the hypothesis that aerenchyma development is an effective strategy in habitats subject to persistent flooding, while elevating activity of enzymes for ethanolic fermentation is effective only under ephemeral flooding. The range of phenotypic responses observed illustrates contrasting adaptive strategies that can lead to habitat isolation and evolutionary divergence. PMID- 21636425 TI - Molecular insights into the taxonomy of Glyceria (Poaceae: Meliceae) in North America. AB - Eighteen Glyceria species grow in the United States and Canada, with 16 being native to the region. We used data from morphology and three chloroplast DNA intergenic regions to address taxonomic questions concerning Glyceria in North America, particularly the status of G. declinata, G. occidentalis, G. fluitans, G. striata, and G. elata in western North America. The chloroplast data confirmed the presence of two European species, G. declinata and G. fluitans, in western North America. Glyceria occidentalis was exceptional among the taxa studied in having chloroplast genotypes that fell into two different clades, one of which contained G. fluitans and the other the North American species G. leptostachya. The morphological data showed G. occidentalis to be intermediate between G. fluitans and G. leptostachya with respect to their distinguishing characters. Based on these results, we hypothesize that G. occidentalis consists of hybrids between G. fluitans and G. leptostachya. Glyceria elata and G. striata, which have sometimes been treated as a single species, had different chloroplast genotypes, supporting their recognition as distinct taxa. DNA data from all three intergenic regions would be needed for unequivocal identification of the non hybrid species examined. PMID- 21636426 TI - Phylogenetic relationships and ecological speciation in the mistletoe Tristerix (Loranthaceae): the influence of pollinators, dispersers, and hosts. AB - Phylogenies can provide valuable information on biotic and abiotic factors associated with speciation. We examined species relationships in Tristerix (Loranthaceae), a genus of 11 species with an Andean distribution from Colombia to Chile. A previous classification divided Tristerix into subgenera Tristerix (two species) and Metastachys (nine species). We tested this classification by generating a molecular phylogeny of the genus using nuclear ribosomal DNA ITS and chloroplast atpB-rbcL intergenic spacer and trnL-F regions. All partitions generally gave congruent trees, thus a combined analysis was conducted. Tristerix was composed of a northern clade (six species) and a southern clade (four species). Tristerix verticillatus and T. penduliflorus (Metastachys) were strongly supported as members of the (southern) subgenus Tristerix clade. Speciation appears to be correlated with the emergence of matorral and cloud forest biomes and is driven by interactions with pollinators and seed dispersers. Tristerix aphyllus is sister to T. corymbosus of the matorral, not to neighboring temperate forest populations, thus rendering the latter species paraphyletic. This ecological speciation event may have occurred in sympatry. Tristerix provides excellent examples of how, during the orography of the Andes, many dynamic and interacting ecological factors have influenced their speciation. PMID- 21636427 TI - Delimitation of major lineages within Cuscuta subgenus Grammica (Convolvulaceae) using plastid and nuclear DNA sequences. AB - Subgenus Grammica, the largest and most diverse group in the parasitic genus Cuscuta, includes 130 species distributed primarily throughout the New World, with Mexico as its center of diversity. To circumscribe the subgenus and assess the relationships among its major lineages, we conducted the first phylogenetic study of Grammica using plastid trnL-F and nrITS sequences from a wide taxonomic sampling covering its morphological, physiological, and geographical diversity. With the exception of one species belonging elsewhere, the subgenus was found to be monophyletic. The results further indicate the presence of 15 well-supported major clades within Grammica. Some of those lineages correspond partially to earlier taxonomic treatments, but the majority of groups are identified in this study for the first time. The backbone relationships among major clades, however, remain weakly supported or unresolved in some cases. The phylogenetic results indicate that the fruit dehiscence character is homoplastic, thus compromising its value as a major taxonomic and evolutionary feature. While several striking cases of long-distance dispersal are inferred, vicariance emerges as the most dominant biogeographical pattern for Cuscuta. Species placed within one of the clades with a predominantly South American distribution are hypothesized to have substantially altered plastid genomes. PMID- 21636428 TI - Gravitropism in the starch excess mutant of Arabidopsis thaliana. AB - Amyloplasts are hypothesized to play a key role in the cellular mechanisms of gravity perception in plants. While previous studies have examined the effects of starch deficiency on gravitropic sensitivity, in this paper, we report on gravitropism in plants with a greater amount of starch relative to the normal wild type. Thus, we have studied the sex1 (starch excess) mutant of Arabidopsis thaliana, which accumulates extra starch because it is defective in a protein involved in the regulation of starch mobilization. Compared to the wild type (WT), sex1 seedlings contained excess starch in cotyledons, hypocotyls, the root hypocotyl transition zone, the body of the root, root hairs, and in peripheral rootcap cells. Sedimented amyloplasts were found in both the WT and in sex1 in the rootcap columella and in the endodermis of stems, hypocotyls, and petioles. In roots, the starch content and amyloplast sedimentation in central columella cells and the gravitropic sensitivity were comparable in sex1 and the WT. However, in hypocotyls, the sex1 mutant was much more sensitive to gravity during light-grown conditions compared to the WT. This difference was correlated to a major difference in size of plastids in gravity-perceiving endodermal cells between the two genotypes (i.e., sex1 amyloplasts were twice as big). These results are consistent with the hypothesis that only very large changes in starch content relative to the WT affect gravitropic sensitivity, thus indicating that wild-type sensing is not saturated. PMID- 21636429 TI - Climatic reconstruction at the Miocene Shanwang basin, China, using leaf margin analysis, CLAMP, coexistence approach, and overlapping distribution analysis. AB - The reconstruction of the climate in the Miocene Shanwang basin is an important link in understanding past climate and environmental changes in East Asia. A recent study showed that the mean annual temperature (MAT) estimates derived from leaf margin analysis (LMA) and the Climate Leaf Analysis Multivariate Program (CLAMP) conflicted with and were remarkably lower than those estimated by the coexistence approach (CA). Overlapping distribution analysis (ODA), a new method introduced here, is used to reconstruct the Shanwang Miocene climate based explicitly on local plant distribution data and associated meteorological stations. The Shanwang flora (17-15.2 Ma) suggests a MAT of 10.9-14.5 degrees C and a mean annual precipitation (MAP) of 1107.3-1880.0 mm. This result is closer to the values derived from CLAMP and LMA than that obtained by CA. This report is the first comprehensive intercomparison of foliar physiognomic and nearest living relative climate proxies in a Chinese context and provides important cross validation of results. PMID- 21636430 TI - Cobbania corrugata gen. et comb. nov. (Araceae): a floating aquatic monocot from the Upper Cretaceous of western North America. AB - The fossil record of aquatic flowering plants broadens our understanding of their former diversity and origins from terrestrial ancestors. This paper describes a floating aquatic monocot from 71 whole plants and several isolated leaf fragments from Upper Cretaceous oxbow lake sediments in the Dinosaur Park Formation, Alberta, Canada. The new material is represented by rosettes of leaves and roots attached to short stems that are interconnected by stolons and corresponds to the fossil aroid originally described as Pistia corrugata Lesquereux. Up to six plants have been found interconnected on a single slab suggesting that these plants grew in extensive floating mats covering lakes and calm stretches of rivers. Stems have up to six leaves and large numbers of branched aquatic roots. The leaf is trumpet-shaped with an elongate clasping petiole, large aerenchymatous base, and a nearly circular blade rim. Leaf bases are often filled with sediment giving the leaf the appearance of having a basal pouch. Petioles have 6-9 veins that divide into an upper and lower set, and veins converge at an apical notch. A submarginal collective vein and at least two marginal veins with branching veins form the leaf rim. A series of dichotomizing and anastomosing veins characterize the adaxial leaf surface. Tertiary and quaternary veins form polygonal areolae. Leaf surfaces are covered in trichomes that, like those in Pistia stratiotes, probably aided in buoyancy. A reconstruction of the plant is presented. Based on unique leaf morphology, these fossil plants are clearly not assignable to the genus Pistia and are described as Cobbania corrugata (Lesquereux) Stockey, Rothwell et Johnson gen. et comb. nov. Recent systematic analyses using molecular characters resolve two separate origins of floating aquatic aroids included in the duckweeds and the genus Pistia. This new fossil genus increases our understanding of colonization of aquatic habitats by revealing a third possible origin of the floating aquatic habit within Araceae. PMID- 21636431 TI - Does Macaronesia exist? Conflicting signal in the bryophyte and pteridophyte floras. AB - Macaronesia, which includes five mid-Atlantic archipelagos (Azores, Madeira, Selvagems, Canaries, and Cape Verdes), has been traditionally recognized as a distinct biogeographic unit whose circumscription has been intimately associated with the hypothesis that the flora is a relict of a formerly broadly distributed subtropical Tertiary flora. The concept of Macaronesia is revisited here using parsimony and Bayesian analyses of floristic data sets for the moss, liverwort, and pteridophyte floras. All analyses reject the monophyly of Macaronesia s.l., resolving the Cape Verdes with tropical Africa. Of the other Macaronesian archipelagos, the liverwort and pteridophyte analyses support, or could not reject, an Azorean-Madeiran-Canarian clade (hereafter Macaronesia s.s.), but the moss analysis resolves the Canary Islands as sister to North Africa, thus rejecting the concept of Macaronesia s.s. for this group. Dynamic interchange of taxa with neighboring continental areas rather than relictualism best explains the relationships of the Cape Verde cryptogamic flora and the Canary Island moss flora. In contrast, relictualism is consistent with a monophyletic Macaronesia s.s. for liverworts and pteridophytes. However, from the limited information available on relationships of endemic cryptogams, this explanation alone may be unsatisfactory. Spatially congruent patterns may, in fact, conceal a complex mixture of relictual distributions and more recent speciation and dispersal events. PMID- 21636432 TI - Dynamics of freeze-thaw embolism in Smilax rotundifolia (Smilacaceae). AB - Freeze-thaw cycles pose a major physiological challenge for all temperate perennial plants, but monocotyledonous vines face a still greater risk because their few large vessels are especially susceptible to embolism and are not replaced by secondary growth. The genus Smilax is particularly remarkable because it is widespread in the tropics but includes species that survive the hard frosts of New England winters. Smilax rotundifolia was monitored for a year for evidence of stem xylem freeze-thaw cavitation and refilling. Embolism of metaxylem was complete by late November and was completely reversed by late April, when root pressures rose as high as 100 kPa. Protoxylem remained full of sap throughout the year in cryogenic scanning electron micrographs. Three methods were used to quantify embolism: percent loss conductivity (PLC), gravimetric air fraction (GAF: mass of water in stem xylem relative to capacity), and cryogenic scanning electron microscopy (cryo-SEM). The three methods corroborated one another well and gave quantitatively similar results. Osmolality of xylem sap extracted from exuding stems was 64 mol/kg (+/-7.0, N = 8), consistent with the root pressures observed. Strong root pressure can account for Smilax's survival in temperate regions with severe frosts, where few monocots with persistent aboveground organs are found. PMID- 21636433 TI - Hawkmoth pollination of aerangoid orchids in Kenya, with special reference to nectar sugar concentration gradients in the floral spurs. AB - The African orchid flora has a high proportion of species with long-spurred white flowers. Few data exist to test the prediction that this floral syndrome pattern reflects an important role for hawkmoth pollination in the evolution and ecology of these orchids. The pollination biology of five aerangoid orchid species (Rangaeris amaniensis, Aerangis brachycarpa, A. confusa, A. thomsonii, and A. kotschyana) was investigated in Kenya. Four of these have long spurs (>10 cm) and were pollinated by Agrius convolvuli and Coelonia fulvinotata. Aerangis confusa, which has relatively short spurs (ca. 4 cm), was pollinated by the short-tongued hawkmoths Hippotion celerio and Daphnis nerii. Nectar frequently filled the entire spur in some of the study species, even at anthesis. Sugar concentration of the nectar of four species was found to vary from ca. 1% at the mouth of the spur to 20% at the tip. Gradients were expressed more strongly in species with long, straight spurs. Species with spirally twisted spurs showed both steep and shallow nectar gradients. These gradients, previously unknown in plants, may function as a "sugar trail," enticing long-tongued hawkmoths to probe deeply into spurs without incurring the cost of filling an entire spur with concentrated nectar. In addition, the most concentrated nectar is kept out of reach of short tongued pollinators. PMID- 21636434 TI - Transfer of glyphosate resistance: evidence of hybridization in Conyza (Asteraceae). AB - Transfer of herbicide resistance genes between crops and weeds is relatively well documented; however, far less information exists for weed-to-weed interactions. The hybridization between the weedy diploids Conyza canadensis (2n = 18) and C. ramosissima (2n = 18) was investigated by monitoring transmission of the allele conferring resistance to N-phosphonomethyl glycine (glyphosate). In a multivariate quantitative trait analysis, we described the phylogenic relationship of the plants, whereas we tested seed viability to assess potential postzygotic reproductive barriers (PZRB) thus affecting the potential establishment of hybrid populations in the wild. When inflorescences were allowed to interact freely, approximately 3% of C. ramosissima or C. canadensis ova were fertilized by pollen of the opposing species and produced viable seeds; >95% of the ova were fertilized under no-pollen competition conditions (emasculation). The interspecific Conyza hybrid ( ) demonstrated an intermediate phenotype between the parents but superior resistance to glyphosate compared to the resistant C. canadensis parent. Inheritance of glyphosate resistance in the selfed ( ) followed the partially dominant nuclear, single-gene model; backcrosses confirmed successful introgression of the resistance allele to either parent. Negligible PZRB were observed in the hybrid progenies, confirming fertility of the C. canadensis * C. ramosissima nothotaxa. The implications of introgressive hybridization for herbicide resistance management and taxonomy of Conyza are discussed. PMID- 21636435 TI - Male-biased nectar production in a protandrous herb matches predictions of sexual selection theory in plants. AB - Nectar production may disproportionately benefit male relative to female pollination success. In such cases, sexual selection is often suggested as the cause of asymmetric benefits, yet sexual selection in plants-particularly plants with hermaphroditic flowers-is infrequently tested empirically. Here, I used a protandrous herb with male-biased nectar production (Chrysothemis friedrichsthaliana, Gesneriaceae) to test predictions from sexual selection theory. During three flowering seasons, I measured nectar production, pollinator visits, and male and female fecundity following different numbers of cross pollination events. In accordance with sexual selection predictions, (1) nectar production was greater during the male phase by at least 65%; (2) visits by the main pollinator (hummingbird Phaethornis striigularis) were limiting for part of the season, indicating that plants had to compete for pollinator visits; (3) pollinators spent 53% more time per visit and made 86% more visits to male- vs. female-phase flowers, suggesting that nectar increased male more than female pollination success; and (4) female fecundity was maximized by one visit, whereas male fecundity continued to increase with additional visits. Autonomous self pollination further reduced visit requirements for maximum female seed set. These findings match specific sexual selection predictions: they link an observable male bias in a secondary sexual trait (nectar) to positive responses of mating participants (pollinators), resulting in more mating opportunities for mate limited males, relative to apparently resource-limited females. This field testing of theoretical predictions provides unique evidence that sexual selection helps maintain nectar production patterns in this and, quite likely, other hermaphroditic plant species. PMID- 21636436 TI - Fertility of Vriesea gigantea Gaud. (Bromeliaceae) in southern Brazil. AB - Plant fertility is a central subject of many questions in plant evolutionary and conservation biology. Pollen availability, abiotic resources, and flowering pattern can limit fruit and seed production. Open pollination and pollen supplementation studies are used to estimate any pollen limitation in natural populations. To study the impact of these factors on the reproductive success of Vriesea gigantea, an epiphytic bromeliad in the Atlantic Rainforest in Brazil, its fertility in four natural populations in Itapua State Park was assessed by considering plant and inflorescence size, flower production, fruit and seed set, flower and fruit set pattern, and seed viability and germination rate. Supplemental pollination in adult plants was used to determine whether fruit production in V. gigantea is limited by reception of pollen. The results showed that V. gigantea has a high production of flowers, fruits, and seeds. Seeds are highly viable in all populations, presenting an average germination rate of 94% (SE +/- 3.5). Plants of V. gigantea from Itapua State Park are highly fertile. The high proportion of fruit and seed set after manual hand pollination indicates that the species is self-compatible. Pollination treatments showed evidence of pollinator limitation in the Itapua State Park population. PMID- 21636437 TI - Chaetomium elatum (Kunze: Chaetomiaceae) as a root-colonizing fungus in avocado: is it a mutualist, cheater, commensalistic associate, or pathogen? AB - Plants support numerous root colonists that may share morphological characteristics with mycorrhizal fungi but may play different roles in the rhizosphere. To determine the function of one such root-colonizing fungus, Chaetomium elatum, the infectivity and composition of inoculum containing C. elatum were varied independently of and in association with the known mutualist Glomus intraradices under two light intensities. Maximum plant benefit occurred with mixtures of both G. intraradices and C. elatum and under high light intensity. Under low light intensity and in monoculture, C. elatum functioned as a weak pathogen that was able to kill host plants. Here, maximum plant mortality was associated with the highest levels of C. elatum infectivity. When G. intraradices was present, no negative impact of C. elatum was detected. Intraspecific interactions were important in predicting sporulation rates for both fungi, whereas no interspecific fungal interactions were detected. In the presence of G. intraradices, C. elatum appears to function as a "commensalistic associate," neither impacting plant growth nor sporulation by G. intraradices. Overall, C. elatum appears to be multifunctional, serving as both a rhizoplane and rhizophere fungus, opportunistically colonizing plant roots and only becoming pathogenic when resources are severely limited and intraspecific competition is high. This multifunctional strategy may be shared with other fungi that form similar structures in roots. PMID- 21636438 TI - Gametophyte ecology and demography of epiphytic and terrestrial tropical ferns. AB - Factors that influence the distribution of ferns are poorly understood and likely reflect the ecology of both the sporophyte and the gametophyte generation. Little study has been done on the ecology of the gametophyte generation, especially in regard to tropical species. The goal of this study was to examine demography and the influence of light and disturbance on the distribution of the gametophytes of several tropical epiphytic, hemiepiphytic, and terrestrial fern species. Through a series of observational and experimental studies, we found that increased terrestrial gametophyte density and richness were related to both increased light and disturbance. By contrast, increased light had no influence, and increased disturbance negatively affected epiphytic density. Over a 25-mo demographic study, epiphytic and hemiepiphytic species had significantly greater longevities and lower recruitment rates than terrestrial species. Such unique strategies may have evolved in response to different disturbance regimens between the two habitats. Terrestrial species encounter and are adapted to more frequent disturbance and have invested in rapid gametophyte growth and recruitment. Epiphytic species may be more influenced by bryophyte competition, and in habitats of relatively low disturbance, they have invested in greater size and longevities. In such systems, gametophytes are able to survive for years waiting for favorable recruitment conditions. PMID- 21636439 TI - Erratum. PMID- 21636440 TI - Wood vessel diameter is related to elevation and genotype in the Hawaiian tree Metrosideros polymorpha (Myrtaceae). AB - We tested the hypothesis that trees growing at high elevations with occasional freezing temperatures have smaller diameter xylem vessels than trees of the same species growing at lower and warmer elevations. The young branch wood of the wide ranging Hawaiian tree species Metrosideros polymorpha (Myrtaceae) was examined in three natural field populations (high, middle, and low elevations: 2469, 1280, and 107 m a.s.l., respectively) and contrasted with seedlings from these populations that were grown in a common garden at middle elevation (1190 m). Previous studies showed that these populations have some genetic differences and have distinctive leaf structure and ecophysiological traits. Vessel diameter was significantly smaller in the high elevation field and common garden plants than in middle elevation plants. However, high elevation vessels were wider in common garden plants compared to field plants, indicating that vessel diameter is determined both by genotype (parental populations) and environment (growing conditions different from those of parents). Reduced vessel diameter has implications for resistance to cavitation induced by freezing and/or drought in plants growing near tree line in Hawaii. PMID- 21636441 TI - Floral organogenesis and floral evolution of the Lecythidoideae (Lecythidaceae). AB - The subfamily Lecythidoideae of Lecythidaceae (Brazil nut family) is a dominant group in neotropical forests, especially those of Amazonia. New World members of the family have large showy flowers that are either polysymmetric or monosymmetric. In this study, floral organogenesis of all 10 neotropical genera was examined using SEM. Our observations of floral development are put into the context of a molecular phylogeny based on sequences of the ndhF and trnL-F genes (Am. J. Bot. 94: 289-301). Floral evolution of the subfamily is explained as having undergone four different levels of complexity in regard to floral symmetry. The basal most genera, Grias and Gustavia, have polysymmetric flowers. At level two, represented only by Couroupita, monosymmetry is established through the expression of abaxial dominance and the development of an androecial hood; at this level, abaxial dominance impacts the perianth and androecium, but not the gynoecium. At the third level, monosymmetry is developed in groups of Couratari and Cariniana domestica; but, in the Allantoma/Cariniana decandra lineage, a reversal back to polysymmetric flowers, resulting from a gradual weakening of abaxial dominance, and the loss of the hood has occurred. Finally, in level four, including Bertholletia, Corythophora, Eschweilera, and Lecythis, monosymmetry is so strongly expressed that the gynoecium is also influenced by abaxial dominance. In this group, the hood is complicated in both structure and function, and the floral axis is changed from straight to slightly inclined. This study demonstrates that the development of floral abaxial dominance is the proximate cause of monosymmetry in the Lecythidoideae. We suggest that monosymmetric flowers are more efficiently pollinated, and therefore the bees and bats that pollinate the monosymmetric flowers in this group are ultimately responsible for the monosymmetry. PMID- 21636442 TI - Structure and development of the upper haustorium in the parasitic flowering plant Cuscuta japonica (Convolvulaceae). AB - The anatomical and ultrastructural development of the haustorium of the Cuscuta japonica, a holoparasitic angiosperm, growing on the host plant Impatiens balsamina was studied. After the shoot tips of light-grown parasite seedlings contacted the host, the upper haustorium (external to the host organ) developed through three main successive stages of the haustorial initials, the meristem, and the endophyte primoridium (EP) within the middle layer of the cortex of the parasite stem. The haustorial initial cells were characterized by abundant starch bearing amyloplasts and mitochondria with an expanded intermembrane space. The meristem cells had numerous large chloroplasts with well-developed thylakoids, reflecting the capability for photosynthesis. Commonly, all three stages of haustorial cells contained conspicuous, large nuclei with enlarged nucleoli and dense cytoplasm including many other organelles, indicating a very active metabolism. In the final stage of upper haustorium development, the meristem cells differentiated into the EP, a host-penetrating tissue. The primordium had smaller file cells at the proximal end and elongate digitate cells at the distal end. The file cells divided actively, while the digitate cells contained abundant chloroplasts, dictyosomes, rough endoplasmic reticulum, and other organelles, suggesting that the EP was cytohistologically well organized for penetration into the host tissue. PMID- 21636443 TI - Reproductive potential of balsam fir (Abies balsamea), white spruce (Picea glauca), and black spruce (P. mariana) at the ecotone between mixedwood and coniferous forests in the boreal zone of western Quebec. AB - The reproductive potentials of balsam fir and white spruce (co-dominants in mixedwood forests) and black spruce (dominant in coniferous forests) were studied to explain the location of the ecotone between the two forest types in the boreal zone of Quebec. Four sites were selected along a latitudinal gradient crossing the ecotone. Cone crop, number of seeds per cone, percentage filled seeds, and percentage germination were measured for each species. Balsam fir and white spruce cone crops were significantly lower in the coniferous than in the mixedwood forest, while black spruce had greater crop constancy and regularity between both forest types. Mast years were more frequent for black spruce than for balsam fir in both forest types (mast year data not available for white spruce). The number of seeds per cone was more related to cone size than to forest type for all species. Black spruce produced more filled seeds in the coniferous forest than balsam fir or white spruce. The sum of growing degree-days and the maximum temperature of the warmest month (both for the year prior to cone production) significantly affected balsam fir cone production. The climate related northward decrease in reproductive potential of balsam fir and white spruce could partly explain the position of the northern limit of the mixedwood forest. This could change drastically, however, as the ongoing climate warming might cancel this competitive advantage of black spruce. PMID- 21636444 TI - Ecophysiological responses of two herbaceous species to prescribed burning, alone or in combination with overstory thinning. AB - The oak-rich deciduous forests of the central Appalachian Mountains of eastern North America have changed significantly since the onset of effective fire suppression early in the 20th century. Those changes have resulted in progressively decreasing light and nutrient supplies to herbaceous perennial understory species. Application of ecological restoration treatments such as reintroduction of frequent dormant-season fire and overstory thinning to pre suppression density often increase light, soil temperature and moisture, and short-term nutrient availability to pre-suppression levels. To persist in this environment, perennial understory herbs must be able to acclimate phenotypically to the very different resource supply combinations present with and without fire suppression. As part of a larger study of the response of the long-lived herbaceous perennials Desmodium nudiflorum and Panicum boscii to ecosystem restoration treatments in Ohio mixed-oak forests, this study examined the ecophysiological effects of prescribed burning (B) and the combination of burning and thinning (T + B) in mixed-oak forests in southern Ohio. Control (C) plants had significantly lower maximum photosynthetic rate (A(max)) than those in the treated plots. The enhancement of A(max) averaged 26.7% and 52.7% in the B and T + B treatments, respectively. Plants from the T + B plots had higher quantum yield, stomatal conductance, and photosynthetic nutrient use efficiency than B and C plants. B plants had greater intrinsic water use efficiency (WUE) than plants in the C or T + B treatments. Light saturation point (LSP), light compensation point (LCP), and "dark" respiration (DR) did not differ among treatments. Photosynthetic parameters did vary significantly between the species, but no significant treatment * species interactions were detected. Our results support the hypothesis that prescribed burning, especially when combined with overstory thinning, in these perennial herbs can result in phenotypic acclimation characterized by enhanced photosynthetic performance. PMID- 21636445 TI - Ecological distribution of homobaric and heterobaric leaves in tree species of Malaysian lowland tropical rainforest. AB - Tree species can generally be classified into two groups, heterobaric and homobaric leafed species, according to whether bundle-sheath extensions (BSEs) are found in the leaf (heterobaric leaf) or not (homobaric leaf). In this study, we study whether the leaf type is related to the growth environment and/or life form type, even in a tropical rain forest, where most trees have evergreen leaves that are generally homobaric. Accordingly, we investigated the distribution of leaf morphological differences across different life forms of 250 tree species in 45 families in a tropical rainforest. In total, 151 species (60%) in 36 families had homobaric leaves, and 99 species (40%) in 21 families had heterobaric leaves. We found that the proportion of heterobaric and homobaric leaf species differed clearly across taxonomic groups and life form types, which were divided into five life form types by their mature tree heights (understory, subcanopy, canopy, and emergent species) and as canopy gap species. Most understory (94%) and subcanopy (83%) species such as Annonaceae had homobaric leaves. In contrast, heterobaric leaf trees appeared more frequently in the canopy species (43%), the emergent species (96%) (such as Dipterocarpaceae), and the canopy gap species (62%). Our results suggest that tree species in the tropical rainforest adapt to spatial differences in the environmental conditions experienced at the mature height of each tree species, such as light intensity and vapor pressure difference, by having differing leaf types (heterobaric or homobaric) because these types potentially have different physiological and/or mechanical functions. PMID- 21636446 TI - An evaluation of mechanisms preventing growth and survival of two native species in invasive Bohemian knotweed (Fallopia xbohemica, Polygonaceae). AB - Restoring native diversity to habitats dominated by invasive plants requires improved understanding of mechanisms that allow introduced plants to retain dominance. We used a factorial transplant experiment to assess whether light limitation, nutrient limitation, or allelopathic interference by Fallopia *bohemica reduces growth or survival of Eupatorium perfoliatum or Acer saccharinum. Increased light improved A. saccharinum performance but had no effect on E. perfoliatum growth. Increased light had no effect on A. saccharinum survival but improved E. perfoliatum survival. Activated carbon addition had short-term benefits allowing E. perfoliatum plants to grow for 4 wk and increased A. saccharinum leaf width but not biomass. Nutrient addition had no beneficial effect on transplants. These results in combination with the outcome of a cutting experiment suggest that F. *bohemica achieves competitive superiority primarily by limiting access to light. Species-specific effects and significant interaction effects particularly of light and activated carbon suggest additional mechanisms. Allelopathic interference or interaction with microbial soil organisms may contribute to the lack of native species in populations of F. *bohemica. Restoration efforts should combine selective F. *bohemica control with soil amendments (such as activated carbon) to assess their potential to facilitate return of native species to invaded habitats. PMID- 21636447 TI - Attenuation of incident light in Galax urceolata (Diapensiaceae): concerted influence of adaxial and abaxial anthocyanic layers on photoprotection. AB - Although anthocyanin coloration in lower (abaxial) leaf cells has been documented for numerous species, the functional significance of this character has not been comprehensively investigated according to habitat or leaf orientation. Here, we demonstrate that abaxial anthocyanin may function as a photoprotectant, similarly to its purported role in upper (adaxial) cells, in leaves vulnerable to high irradiance incident on abaxial surfaces. Spectral scans were derived for Galax urceolata leaves with the following phenotypes: abaxial or adaxial anthocyanin only, abaxial and adaxial anthocyanin, and no anthocyanin. To determine whether anthocyanins conferred protection from photoinhibition, maximum photosystem II efficiencies of red (anthocyanic) and green (acyanic) surfaces were compared during and after exposure to photoinhibitory conditions. Leaves were either positioned with their adaxial surfaces facing the light source or inverted to expose abaxial surfaces. Spectral scans showed increased absorptance of 500-600 nm wavelengths by red surfaces (consistent with the absorbance spectrum of anthocyanin), regardless of whether that surface was abaxial or adaxial. Leaves with anthocyanin in either illuminated surface were also photoinhibited less than leaves lacking anthocyanin in that surface. These results suggest that anthocyanic layers reduce absorbed sunlight in the mesophyll not only for adaxial surfaces, but also for the abaxial. Adaxial/abaxial anthocyanin plasticity may therefore be adaptive in high-light environments or during light-sensitive developmental stages where leaf orientation and/or substrate albedo are variable. PMID- 21636448 TI - Secondary phloem anatomy of Cycadeoidea (Bennettitales). AB - Secondary phloem anatomy of several species of Cycadeoidea is described from trunks in the Wieland Collection, Peabody Museum of Natural History. The trunks were collected from the Lakota Formation, Lower Cretaceous, Black Hills of South Dakota. Secondary phloem is extensively developed and consists of alternating, tangential bands of fibers and sieve elements, with rare phloem parenchyma. Uniseriate rays, 2-22 cells high, occur between every one to three files of the axial system. Fibers are long, more than 1200 MUm, approximately 26.6-34.2 MUm in diameter, and have slit-like apertures on the lateral walls. Sieve elements range from 16-25 MUm in diameter and are up to 500 MUm long. Elliptical sieve areas appear on both end and radial walls and measure 10 MUm across; minute spots, which may represent sieve pores, are present within the sieve areas. Secondary phloem of North American Cycadeoidea is similar in organization (alternating tangential bands) and cell types (sieve cells, fibers, axial parenchyma) to that known in other extant and fossil cycadophytes and some seed ferns. The unusual pattern of cell types and thickness of secondary phloem is discussed in the context of plant habit, phloem efficiency, and potential phylogenetic importance. PMID- 21636449 TI - Assessing genetic stability of a range of terrestrial microalgae after cryopreservation using amplified fragment length polymorphism (AFLP). AB - Cryopreservation is the long-term, indefinite storage of living biological resources at ultralow temperatures. It is almost universally assumed that cryogenic storage supports genetic and phenotypic stability of organisms. However, certain components of the cryopreservation process, particularly some cryoprotective additives (CPAs) and free radical mediated cryoinjury, may potentially cause genetic alterations. Genetic integrity in cryopreserved microalgae was assessed using a very sensitive molecular fingerprinting technique, AFLP, on 28 terrestrial microalgal strains. In about half of all investigated strains the AFLP fingerprints revealed, with high levels of reproducibility, clearly detectable genomic differences after cryopreservation employing a widely used standard two-step cooling protocol. Differences ranged from a single fragment position to multiple fragment changes and were compared to differences found between wild-type and UV-light- or radioisotope-induced mutants of Parachlorella kessleri. The basis of the changes are discussed in terms of their reversibility, as may be the case if they are attributed to DNA methylation and/or whether they are true mutations that may potentially manifest in the phenotype. The possibility that cryopreservation selects for genotypically different subpopulations of microalgae is also considered. PMID- 21636450 TI - Drought response of two Mexican oak species, Quercus laceyi and Q. sideroxyla (Fagaceae), in relation to elevational position. AB - To investigate the relationship between the altitudinal distribution of Quercus laceyi and Q. sideroxyla and their physiological responses to drought, we measured relative water content (RWC), water potentials (Psi(predawn) and Psi(midday)), photosynthesis (A(max)), stomatal conductance (g), chlorophyll fluorescence (F(v)/F(m)), and spectral reflectance (400-1100 nm) five times during a 7 wk acute drought. Quercus laceyi was drought tolerant, while Q. sideroxyla was a drought avoider; Q. laceyi tolerated lower RWC (Q. sideroxyla = 54%, Q. laceyi = 44%), Psi(pd) (Q. sideroxyla = -2.6 MPa, Q. laceyi = -3.3 MPa), and Psi(md) (Q. sideroxyla = -4.5 MPa, Q. laceyi = -6.6 MPa). The F(v)/F(m) also declined first in Q. sideroxyla in wk 6, whereas F(v)/F(m) did not decline in Q. laceyi until wk 7. A(max) and g fell in wk 4, 6, and 7 in drought seedlings of both species, suggesting a decline in CO(2) assimilation during the drought. Leaf spectral reflectance increased with time in response to decreases in leaf photosynthetic pigment concentrations in latter weeks of the drought. The results suggest a close association between the altitudinal distributions of these species and their adaptation to water stress. PMID- 21636451 TI - Elevated temperatures increase leaf senescence and root secondary metabolite concentrations in the understory herb Panax quinquefolius (Araliaceae). AB - The response of understory species to elevated temperatures is not well understood but is important because these plants are highly sensitive to their growth conditions. Three-year-old plants of Panax quinquefolius, an understory herb endemic to the eastern deciduous forests of North America, were grown in a greenhouse at 25/20 degrees C (day/night) or 30/25 degrees C for one growing season and analyzed each month. Plants grown at high temperatures had an early onset of leaf senescence and therefore accumulated less carbon. From May to July, P. quinquefolius grown at high temperatures had decreased photosynthesis (52%), stomatal conductance (60%), and root and total biomass (33% and 28%, respectively) compared to plants grown at low temperatures. As P. quinquefolius prepared to overwinter, plants grown at high temperatures had less root biomass (53%) than plants in low temperatures. The amount of storage-root ginsenosides was unaffected by temperature, and differences in storage root size may explain why plants grown at high temperatures had greater concentrations of storage root ginsenosides (49%) than plants grown at low temperatures. Panax quinquefolius is clearly sensitive to a 5 degrees C increase in temperature, and therefore other understory species may be negatively impacted by future increases in global temperature. PMID- 21636452 TI - Population structure of Pacific Cordyline fruticosa (Laxmanniaceae) with implications for human settlement of Polynesia. AB - The Polynesian-introduced Cordyline fruticosa is used as a proxy for reconstructing human colonization patterns in Oceania. Because of its material, nutritional, medicinal, and religious importance, green-leaved C. fruticosa was transferred by Polynesian settlers to virtually every habitable Pacific island before European contact. Previous studies propose that green-leaved C. fruticosa is unable to reproduce sexually. To confirm sterility, crosses between fertile and putatively sterile forms were performed. To look for population structure in C. fruticosa that might confirm sterility as well as illustrate patterns of human migration, amplified fragment length polymorphism data were generated. Genotypic similarities were visualized using neighbor joining phenograms and analyses of molecular variance and principal components. The results from greenhouse crosses show that the Eastern Polynesian form is sterile; this finding is corroborated by a lack of genetic variability in Eastern Polynesian accessions. Sterile C. fruticosa appears to have been preferentially transferred throughout Eastern Polynesia; selection for the sterile form may be related to consumption of its rhizomes. Identification of a sterile form of C. fruticosa, possibly developed within Western Polynesia, may be significant to the systematics of Cordyline because it raises the possibility that the fertile form may actually be native to some Pacific islands. PMID- 21636453 TI - Variation in ovule and seed size and associated size-number trade-offs in angiosperms. AB - Unlike pollen and seed size, the extent and causes of variation in ovule size remain unexplored. Based on 45 angiosperm species, we assessed whether intra- and interspecific variation in ovule size is consistent with cost minimization during ovule production or allows maternal plants to dominate conflict with their seeds concerning resource investment. Despite considerable intraspecific variation in ovule volume (mean CV = 0.356), ovule production by few species was subject to a size-number trade-off. Among the sampled species, ovule volume varied two orders of magnitude, whereas seed volume varied four orders of magnitude. Ovule volume varied positively among species with flower mass and negatively with ovule number. Tenuinucellate ovules were generally larger that crassinucellate ovules, and species with apical placentation (which mostly have uniovulate ovaries) had smaller ovules than those with other placentation types. Seed volume varied positively among species with fruit mass and seed development time, but negatively with seed number. Seeds grew a median 93-fold larger than the ovules from which they originated. Our results provide equivocal evidence that selection minimizes ovule size to allow efficient resource allocation after fertilization, but stronger evidence that ovule size affords maternal plants an advantage in parent-offspring conflict. PMID- 21636454 TI - Is Cycas revoluta (Cycadaceae) wind- or insect-pollinated? AB - Among the Cycadales (Cycadaceae and Zamiaceae), the Zamiaceae are known to be insect-pollinated. In contrast, the Cycadaceae are still considered wind pollinated, although some doubt has been cast on several species, including Cycas revoluta. Using a large population of C. revoluta on Yonaguni Island (Okinawa, Japan), we performed exclusion experiments, documented insects from male and female cones, and analyzed the morphology of the apical part of the ovule to determine the pollination method of this species. Insect exclusion resulted in a notable reduction in seed set, except in a few individuals growing near male cones. The amount of airborne pollen was abundant within a 2-m radius of male cones but decreased markedly beyond this distance. Pollen grains of C. revoluta were found on the body of Carpophilus chalybeus (Nitidulidae, Coleoptera), one of a few species of insects collected from both male cones and female cones far from males. We conclude that C. revoluta relies on both wind (anemophily) and insect pollination (entomophily), although such anemophily is restricted to female trees growing within a 2-m radius of male trees. The nitidulids are not host specific to this cycad and primarily feed on plant tissue but serve as pollinators during pollen release. Cycas revoluta appears to be in an initial mode of animal pollination, as opposed to the host-specific insect pollination observed in most Zamiaceae. PMID- 21636455 TI - Molecular phylogeny of Nyctaginaceae: taxonomy, biogeography, and characters associated with a radiation of xerophytic genera in North America. AB - The four o'clock family (Nyctaginaceae) has a number of genera with unusual morphological and ecological characters, several of which appear to have a "tendency" to evolve repeatedly in Nyctaginaceae. Despite this, the Nyctaginaceae have attracted little attention from botanists. To produce a phylogeny for the Nyctaginaceae, we sampled 51 species representing 25 genera (of 28-31) for three chloroplast loci (ndhF, rps16, rpl16, and nrITS) and included all genera from North America. Parsimony, likelihood, and Bayesian methods were used to reconstruct the phylogeny for the family. The family is neotropical in origin. A radiation of woody taxa unites Pisonia and Pisoniella with the difficult tropical genera Neea and Guapira, which also form a clade, though neither appears to be monophyletic. This group is sister to a clade containing Bougainvillea, Belemia, and Phaeoptilum. A dramatic radiation of genera occurred in the deserts of North America. The tribe Nyctagineae and its subtribes are paraphyletic, due to over reliance on a few homoplasious characters, i.e., pollen morphology and involucre presence. Two notable characters associated with the desert radiation are cleistogamy and edaphic endemism on gypsum soils. We discuss evolutionary trends in these traits in light of available data about self-incompatibility and gypsum tolerance in Nyctaginaceae. PMID- 21636456 TI - A molecular phylogeny of scaly tree ferns (Cyatheaceae). AB - Tree ferns recently were identified as the closest sister group to the hyperdiverse clade of ferns, the polypods. Although most of the 600 species of tree ferns are arborescent, the group encompasses a wide range of morphological variability, from diminutive members to the giant scaly tree ferns, Cyatheaceae. This well-known family comprises most of the tree fern diversity (~500 species) and is widespread in tropical, subtropical, and south temperate regions of the world. Here we investigate the phylogenetic relationships of scaly tree ferns based on DNA sequence data from five plastid regions (rbcL, rbcL-accD IGS, rbcL atpB IGS, trnG-trnR, and trnL-trnF). A basal dichotomy resolves Sphaeropteris as sister to all other taxa and scale features support these two clades: Sphaeropteris has conform scales, whereas all other taxa have marginate scales. The marginate-scaled clade consists of a basal trichotomy, with the three groups here termed (1) Cyathea (including Cnemidaria, Hymenophyllopsis, Trichipteris), (2) Alsophila sensu stricto, and (3) Gymnosphaera (previously recognized as a section within Alsophila) + A. capensis. Scaly tree ferns display a wide range of indusial structures, and although indusium shape is homoplastic it does contain useful phylogenetic information that supports some of the larger clades recognised. PMID- 21636457 TI - Within-population genetic structure and clonal diversity of a threatened endemic metallophyte, Viola calaminaria (Violaceae). AB - We studied the within-population genetic structure and the clonality extent of Viola calaminaria, a rare endemic species of calamine soils, by means of RAPD markers in two populations (one recent and one ancient) with expected harsh and heterogeneous heavy-metal stress. At a very local scale (0.2-3 m), clonal propagation was detected in both populations, but the levels of clonal diversity were high (number of genets/number of ramets sampled = 0.9 [recent] and 0.76 [ancient]) and the maximal observed extension of the clones was 0.4 m. This indicated that clonality is not, for the species, an important mode of propagation and that clonal growth cannot be interpreted as a strategy for propagating or perpetuating adapted genotypes under harsh ecological constraints. Spatial autocorrelations revealed a significant (P < 0.001) negative value of correlogram slope in the two populations even when a single individual per clone was considered (i.e., analysis at the genet level). We conclude that spatial genetic structure at a very local scale reflects limited gene flow due to restricted seed dispersal rather than variation in clonal pattern in response to environmental heterogeneity. At a larger scale (2-30 m), spatial autocorrelations revealed a positive (P < 0.001) correlation at < 3 m and a random pattern at larger distances for the two populations. This suggested a patchy distribution of the genetically linked individuals associated with a disrupted pattern at a longer distance probably due to gene flow by pollen dispersal and a seed bank effect. The implications for the conservation of V. calaminaria are discussed. PMID- 21636458 TI - Self-pollen interference is absent in wild radish (Raphanus raphanistrum, Brassicaceae), a species with sporophytic self-incompatibility. AB - Explaining the diversity of mating systems and floral forms in flowering plants is a long-standing concern of evolutionary biologists. One topic of interest is the conditions under which self-pollination can interfere with seed set for flowering plants with a self-incompatibility system. We investigated the effect of self-pollen interference for wild radish, Raphanus raphanistrum, which has sporophytic self-incompatibility. We performed pollinations and determined seed set for plants grown in the greenhouse, using pollen mixtures representing either self- with outcross-pollen or outcross-pollen alone. Stigmas were collected for a subset of pollinated flowers to determine the number of pollen grains applied. Average seed set for the self/cross (5.13 seeds/pollination) and cross treatments (5.09 seeds/pollination) did not differ significantly. Stigmatic pollen loads averaged around 700 grains, an amount close to observed natural pollen loads on R. raphanistrum. We concluded that for R. raphanistrum in natural populations, self-pollen is unlikely to interfere with outcross-pollen success. This study is the first to investigate effects of self-pollen interference on seed set for a homomorphic species with sporophytic self-incompatibility where rejection occurs at the stigmatic surface. PMID- 21636459 TI - Seed size, seedling morphology, and response to deep shade and damage in neotropical rain forest trees. AB - To investigate the existence of coordinated sets of seedling traits adapted to contrasting establishment conditions, we examined evolutionary convergence in seedling traits for 299 French Guianan woody plant species and the stress response in a shadehouse of species representing seed size gradients within five major cotyledon morphology types. The French Guianan woody plant community has larger seeds than other tropical forest communities and the largest proportion of hypogeal cotyledon type (59.2%) reported for tropical forests. Yet the community includes many species with intermediate size seeds that produce seedlings with different cotyledonal morphologies. A split-plot factorial design with two light levels (0.8% and 16.1% PAR) and four damage treatments (control, seed damage, leaf damage, stem damage) was used in the shadehouse experiment. Although larger seeded species had higher survival and slower growth, these patterns were better explained by cotyledon type than by seed mass. Even larger-seeded species with foliar cotyledons grew faster than species with reserve-type cotyledons, and survival after stem grazing was five times higher in seedlings with hypogeal cotyledons than with epigeal cotyledons. Thus, to predict seedling performance using seed size, seedling morphology must also be considered. PMID- 21636460 TI - Reaction tissue formation and stem tensile modulus properties in wild-type and p coumarate-3-hydroxylase downregulated lines of alfalfa, Medicago sativa (Fabaceae). AB - To our knowledge, xylary reaction tissue has never been reported in a forage crop species. Here we report the discovery of reaction tissue in a transgenic line of Medicago sativa (pC3H, for the gene for p-coumarate-3-hydroxylase) with reduced lignin content and in the wild-type (WT) line. Based on microscopy and biomechanical testing of internodal alfalfa branch sections, the transgenic (pC3H I) line, relative to the WT (1) apparently formed more reaction tissue containing gelatinous fibers with adjacent thick-walled fibers (presumed to be "intermediate" tissue) more rapidly, (2) had more xylem tissue, and (3) had comparable tensile dynamic modulus properties. These findings thus establish the (limited) ability of this perennial angiosperm to form (inducible) reaction tissue in a manner somewhat analogous to that of woody arborescent angiosperms. The potential of effectuating reductions in lignin amounts in (woody) angiosperms with increased formation of reaction (tension wood) tissue is discussed because reaction tissues are often viewed as a deleterious trait in processing for many agronomic/industrial applications, especially with the current interest in biofuels. PMID- 21636461 TI - Characterization of Rubisco activase from thermally contrasting genotypes of Acer rubrum (Aceraceae). AB - The lability of Rubisco activase function is thought to have a major role in the decline of leaf photosynthesis under moderate heat (<35 degrees C). To investigate this further, we characterized Rubisco activase and explored its role in the previously demonstrated thermal acclimation and inhibition of two genotypes of Acer rubrum originally collected from Florida (FL) and Minnesota (MN). When plants were grown at 33/25 degrees C (day/night) for 21 d, the FL genotype compared to the MN genotype maintained about a two-fold increase in leaf photosynthetic rates at 33-42 degrees C and had a 22% increase in the maximal rate of Rubisco carboxylation (V(cmax)) at 33 degrees C under nonphotorespiratory conditions. Both genotypes had two leaf Rca transcripts, likely from equivalent alternative splicing events. The RCA1 and RCA2 proteins increased modestly in FL plants under warmer temperature, while only RCA2 protein increased in MN plants. Rubisco large subunit (RbsL) protein abundance was relatively unaffected in either genotype by temperature. These results support the idea that Rubisco activase, particularly the ratio of Rubisco activase to Rubisco, may play a role in the photosynthetic heat acclimation in A. rubrum and may have adaptive significance. This mechanism alone is not likely to entirely explain the thermotolerance in the FL genotype, and future research on adaptive mechanisms to high temperatures should consider activase function in a multipathway framework. PMID- 21636462 TI - Genealogy and fine mapping of obscuravenosa, a gene affecting the distribution of chloroplasts in leaf veins, and evidence of selection during breeding of tomatoes (Lycopersicon esculentum; Solanaceae). AB - In the processes of plant domestication and variety development, some traits are under direct selection, while others may be introduced by indirect selection or linkage. In the cultivated tomato (Lycopersicon esculentum = Solanum lycopersicum), and all other Solanaceae examined, chloroplasts are normally absent from subepidermal and mesophyll cells surrounding the leaf veins, and thus, veins appear clear upon subillumination. The tomato mutant obscuravenosa (obv), in contrast, contains chloroplasts in cells around the vein, and thus, veins appear as dark as the surrounding leaf tissue. Among tomato cultivars, the obv allele is common in processing varieties bred for mechanical harvest, but is otherwise rare. We traced the source of obv in processing tomatoes to the cultivar Earliana, released in the 1920s. The obv locus was mapped to chromosome 5, bin 5G, using introgression lines containing single chromosome segments from the wild species L. pennellii. This region also contains a quantitative trait locus (QTL) for plant height, pht5.4, which cosegregated with SP5G, a paralog of self-pruning (sp), the gene that controls the switch between determinate and indeterminate growth in tomato. The pht5.4 QTL was partially dominant and associated with a reduced percentage of red fruit at harvest. Our data suggest that the prevalence of obv in nearly all processing varieties may have resulted from its tight linkage to a QTL conferring a more compact, and horticulturally desirable, plant habit. PMID- 21636463 TI - Molecular and morphological evidence reveals introgression in swarms of the invasive taxa Fallopia japonica, F. sachalinensis, and F. xbohemica (Polygonaceae) in the United States. AB - Fallopia japonica (Japanese knotweed, Polygonaceae) is a well-known East Asian perennial that is established throughout the U.S. and Europe. Another congener, F. sachalinensis, and their hybrid, F. *bohemica, also persist on both continents. Their invasive success is primarily attributed to their ability to spread via clonal growth. However, mounting evidence suggests invasion history and dynamics differ between continents and that sexual reproduction is more common than previously assumed. We used published morphological traits designed to distinguish the three taxa to characterize their distribution in 24 New England towns. We found continuous variation of all five traits, with 84% of our 81 individuals having at least one trait outside parental limits. Hierarchical cluster analysis, along with two chloroplast and one nuclear species-specific markers, suggests the presence of intercrossing, segregating hybrids, and likely introgression between F1 hybrids and F. japonica. Our markers also show the first evidence of bidirectional hybridization between parental taxa in the U.S., emphasizing the complex structure of populations in our region. This study is a first step toward unraveling the evolutionary forces that have made these taxa such aggressive invaders in the U.S. The data may also affect management strategies originally designed for largely monomorphic, clonal populations. PMID- 21636464 TI - Genetic diversity and clonal vs. sexual reproduction in Fallopia spp. (Polygonaceae). AB - Although fundamental to the study of invasion mechanisms, the relationship between mode of reproduction and plant invasion is not well understood. Fallopia japonica (Japanese knotweed), a highly aggressive invasive plant in both Europe and North America, serves as a model species for examining this relationship. In Britain, F. japonica var. japonica is a single female clone reproducing solely through vegetative growth or obligate hybridization with other Fallopia spp. In the U.S., however, there is more evidence for sexual reproduction. Here, simple sequence repeat (SSR) markers were developed, and three Massachusetts populations were sampled at regular intervals. The amount of sexual and clonal reproduction in each population was determined based on within-population genetic diversity. Clonal growth was apparent, but the populations together contained 26 genotypes and had evidence of sexual reproduction. One genotype that was present in all populations matched the single aggressive British clone of F. japonica var. japonica. Also, a potentially diagnostic marker for the F. sachalinensis genome provided evidence of inter- and intraspecific sexual reproduction and introgression. These differences observed in U.S. populations compared to European populations have significant implications for management of Fallopia spp. in the U.S. and underscore the importance of regional studies of invasive species. PMID- 21636465 TI - Patterns of variation of a common fern (Athyrium filix-femina; Woodsiaceae): population structure along and between altitudinal gradients. AB - Genetic variability of Athyrium filix-femina populations was evaluated with regard to phenotypic, allozyme, and RAPD variation in 20 Swiss populations along five altitudinal gradients at four different elevations in the northern Swiss Alps. Additionally, allozyme and phenotypic variations in one Italian and two Spanish populations were compared with the variation in the Swiss populations. We hypothesized that there will be statistically significant genetic differences among populations of different altitudes and sites. The results showed no substantial correlation between genetic variation and phenotypic variation among Swiss populations. These results imply that outbreeding and effective gene exchange (long-distance spore dispersal) are the keys to population structure in this fern species, and as a consequence, phenotypic plasticity is assumed to be favored. This contrasts with results found in similar studies of herbaceous flowering plants where genetic adaptation to gradients like altitude is common. However, when data from the more distant Italian and Spanish populations of A. filix-femina were included, significant variation was detected. PMID- 21636466 TI - The evolution of distyly from tristyly in populations of Oxalis alpina (Oxalidaceae) in the Sky Islands of the Sonoran Desert. AB - The evolution of distyly from tristyly was investigated in populations of Oxalis alpina at high elevations throughout the Sky Islands of the Sonoran Desert. Incompatibility systems in tristylous populations, where self-incompatible short , mid-, and long-styled morphs occur in populations, vary from those typical of tristylous species in which each morph is equally capable of fertilizing ovules of the other two morphs, to breeding systems in which incompatibility relationships are asymmetric. In these populations, selection against the allele controlling expression of the mid-styled morph is likely. The degree of modification of incompatibility in the short- and long-styled morphs in 10 populations was strongly associated with fewer mid-styled morphs, supporting models predicting the effect of these modifications of incompatibility on frequency of the mid-styled morph. Self-compatibility of the mid-styled morph may be important for maintaining the frequency of this morph, depending on the level of self-pollination, self-fertilization, and the extent of inbreeding depression. Modifications of incompatibility in tristylous populations and the distribution of distylous populations of O. alpina in the Sky Island region have similar geographic components, indicating the potential importance of historical factors in the evolution of distyly from tristyly. PMID- 21636467 TI - Do antheridiogens act via gametophyte size? A study of Woodwardia radicans (Blechnaceae). AB - For many plants, sex is not fixed by genotype but determined by environmental conditions during development. In homosporous pteridophytes, sex is environmentally determined by the presence or absence of antheridiogens, maleness inducing pheromones. It has been proposed that antheridiogens primarily reduce growth rate, with small gametophyte size responsible for maleness. To test this hypothesis, the effects of antheridiogen and intergametophytic competition on gender expression and gametophyte size were studied in a culture experiment with Woodwardia radicans. We found that (1) antheridiogen inhibited growth of gametophytes; and (2) slow growth favored maleness, whereas fast growth favored femaleness, irrespective of the presence or absence of antheridiogen. Both conclusions are consistent with the hypothesis that, in W. radicans, antheridiogen effect is mediated by size. They also agree with the "size advantage" hypothesis in which energetic limitations associated with relatively small individual size impose a less severe limitation for male reproductive success than for female reproductive success. The results are also discussed with regard to a genetic sex-determining pathway that has recently been identified. PMID- 21636468 TI - Increased selfing and decreased effective pollen donor number in peripheral relative to central populations in Picea sitchensis (Pinaceae). AB - Because mating system can be influenced by effective neighborhood size, density, and isolation, populations at range peripheries may differ from those in the center. The importance of peripheral populations to conservation and evolution is controversial, and additional information about their genetic structure and evolutionary dynamics will inform conservation strategies. In wind-pollinated species, selfing rate is generally negatively correlated with population size and density, and inbreeding may therefore increase toward range peripheries. Picea sitchensis has a long and narrow range along the Pacific Coast of North America that tapers toward the northern and southern peripheries. We investigated whether central and peripheral populations differ in mating system parameters. The results suggest that population position within the range has a strong effect on mating system, and geographic isolation appears to be associated with higher selfing. The estimated effective number of pollen donors was much higher in the center of the range (mean = 18.5) than at the periphery (mean = 3.6), while selfing rate increased from 7.3% in central populations to as high as 35.2% in the northern, isolated population. These strong geographical patterns suggest mating system is influenced by both population size and isolation at range peripheries. PMID- 21636469 TI - Carica papaya (Caricaceae): a case study into the effects of domestication on plant vegetative growth and reproduction. AB - Few studies have quantitatively evaluated the gender specific effects of cultivation on plant growth and reproduction. The availability of cultivated and wild populations of different genders of Carica papaya L. (Caricaceae) on Guam provided an opportunity to study these effects quantitatively. We compared the gender specific allometry of height vs. basal stem diameter (H vs. D), stem slenderness ratio (H/D), and the height at first flowering (H(fl)) of carpellate and staminate plants growing under natural conditions (N = 150 each) with those of carpellate and hermaphroditic plants (N = 250 each) from two cultivars (Sunrise and Tainung 2). These comparisons indicated that (1) wild carpellate and staminate plants are significantly taller than either gender of the two cultivars with equivalent D; (2) the scaling exponent governing the H vs. D relationship of both genders of wild plants is significantly higher than that of either cultivated gender; (3) cultivar type does not affect the H vs. D exponent, but gender expression does; (4) gender expression (but not cultivar-type) also affects H(fl) (cultivation substantially reduces carpellate plant H(fl)); and (5) the onset of sexual maturity is associated with a dramatic reversal in H/D ontogeny. Cultivation therefore has "condensed" patterns of vegetative growth in a gender specific manner, whereas gender expression alters both vegetative and reproductive growth significantly more so than does cultivar-type. PMID- 21636470 TI - Ancient paralogy in the cpDNA trnL-F region in Annonaceae: implications for plant molecular systematics. AB - The plastid trnL-F region has proved useful in molecular phylogenetic studies addressing diverse evolutionary questions from biogeographic history to character evolution in a broad range of plant groups. An important assumption for phylogenetic reconstruction is that data used in combined analyses contain the same phylogenetic signal. The trnL-F region is often used in combined analyses of multiple chloroplast markers. These markers are assumed to contain congruent phylogenetic signal due to lack of recombination. Here we show that trnL-F sequences display a phylogenetic signal conflicting with that of other chloroplast markers in Annonaceae, and we demonstrate that this conflict results from ancient paralogy. TrnL-F copy 2 diverged from trnL-F copy 1 (as used in family-wide phylogenetic analyses) in a direct ancestor of the Annonaceae. Although this divergence dates back 88 million years or more, the exons of both copies appear to be intact. In this case, assuming that (putative) chloroplast markers contain the same phylogenetic signal results in an incorrect topology and an incorrect estimate of ages. Our study demonstrates that researchers should be cautious when interpreting gene phylogenies, irrespective of the genome from which they are presumed to have been sampled. PMID- 21636471 TI - Molecular phylogeny, biogeography, and systematics of Dicerandra (Lamiaceae), a genus endemic to the southeastern United States. AB - Dicerandra, an endemic mint of the southeastern United States, comprises nine species, all of which are threatened or endangered and restricted to sandhill vegetation and a mosaic of scrub habitats. Molecular phylogenetic analyses of Dicerandra based on data from the nuclear and plastid genomes for all 13 taxa of the genus, identified two strongly supported clades, corresponding to the four annual and to the five perennial species of Dicerandra. However, the nuclear and plastid trees were incongruent in their placement of two perennial taxa, D. cornutissima and D. immaculata var. savannarum, perhaps due to ancient hybridization or to lineage sorting. Based on these analyses, the widespread D. linearifolia is not monophyletic, with populations of D. linearifolia var. linearifolia falling into either western or eastern clades. The western clade, comprising populations of D. linearifolia var. linearifolia and var. robustior, occurs in an area drained by rivers flowing toward the Gulf of Mexico, whereas the eastern clade, comprising populations of D. linearifolia var. linearifolia, D. densiflora, D. odoratissima, and D. radfordiana (i.e., all the annual species), occupies a region drained by rivers flowing to the Atlantic Ocean. Although this pattern of genetic differentiation between populations from these two river drainages has been documented in several animal species, it has not previously been reported for plants. A revised subgeneric classification is presented to reflect the annual and perennial clades. PMID- 21636472 TI - Phylogeny and biogeography of the sandalwoods (Santalum, Santalaceae): repeated dispersals throughout the Pacific. AB - Results of the first genus-wide phylogenetic analysis for Santalum (Santalaceae), using a combination of 18S-26S nuclear ribosomal (ITS, ETS) and chloroplast (3' trnK intron) DNA sequences, provide new perspectives on relationships and biogeographic patterns among the widespread and economically important sandalwoods. Congruent trees based on maximum parsimony, maximum likelihood, and Bayesian methods support an origin of Santalum in Australia and at least five putatively bird-mediated, long-distance dispersal events out of Australia, with two colonizations of Melanesia, two of the Hawaiian Islands, and one of the Juan Fernandez Islands. The phylogenetic data also provide the best available evidence for plant dispersal out of the Hawaiian Islands to the Bonin Islands and eastern Polynesia. Inability to reject rate constancy of Santalum ITS evolution and use of fossil-based calibrations yielded estimates for timing of speciation and colonization events in the Pacific, with dates of 1.0-1.5 million yr ago (Ma) and 0.4-0.6 Ma for onset of diversification of the two Hawaiian lineages. The results indicate that the previously recognized sections Polynesica, Santalum, and Solenantha, the widespread Australian species S. lanceolatum, and the Hawaiian species S. freycinetianum are not monophyletic and need taxonomic revision, which is currently being pursued. PMID- 21636473 TI - Mannitol-enhanced, fluid-phase endocytosis in storage parenchyma cells of celery (Apium graveolens; Apiaceae) petioles. AB - We recently demonstrated the occurrence of a sucrose-enhanced, fluid-phase endocytic (FPE) mechanism of nutrient uptake in heterotrophic cells. In the present work, the possible enhancement/induction of FPE by photoassimilates other than sucrose was investigated by measuring the incorporation of the fluorescent endocytosis marker d-TR (dextran-Texas red, 3000 mw) into celery (Apium graveolens) petiole storage parenchyma (CSP), a tissue that transports and accumulates mannitol. Mannitol uptake in these cells is biphasic, with a hyperbolic phase at concentrations below 20 mM and a linear phase above 20 mM external solute concentration. In the absence of mannitol, or in its presence at concentrations within the hyperbolic phase, CSP cells accumulated low levels of d TR. Conversely, d-TR accumulation by CSP cells was greatly enhanced in the presence of mannitol at concentrations within the linear phase. At high external mannitol concentration, d-TR accumulation was prevented by the endocytic inhibitors LY294002 and latrunculin B. In addition, d-TR uptake was temperature dependent under high mannitol concentration. Microscopic observations revealed that d-TR accumulated in the vacuole. These data support the occurrence of an FPE mechanism in CSP cells that participates in trapping and transport of photoassimilates to the vacuole. The FPE mechanism is enhanced by high mannitol concentrations. PMID- 21636474 TI - Calcium function and distribution during fertilization in angiosperms. AB - Calcium has an essential signaling, physiological, and regulatory role during sexual reproduction in flowering plants; elevation of calcium amounts is an accurate predictor of plant fertility. Calcium is present in three forms: (1) covalently bound calcium, (2) loosely bound calcium typically associated with fixed and mobile anions (ionic bonding); and (3) cytosolic free calcium-an important secondary messenger in cell signaling. Pollen often requires calcium for germination. Pollen tube elongation typically relies on external calcium stores in the pistil. Calcium establishes polarity of the pollen tube and forms a basis for pulsatory growth. Applying calcium on the tip may alter the axis; thus calcium may have a role in determining the directionality of tube elongation. In the ovary and ovule, an abundance of calcium signals receptivity, provides essential mineral nutrition, and guides the pollen tube in some plants. Calcium patterns in the embryo sac also correspond to synergid receptivity, reflecting programmed cell death in one synergid cell that triggers degeneration and prepares this cell to receive the pollen tube. Male gametes are released in the synergid, and fusion of the gametes requires calcium, according to in vitro fertilization studies. Fusion of plant gametes in vitro triggers calcium oscillations evident in both the zygote and primary endosperm during double fertilization that are similar to those in animals. PMID- 21636475 TI - Do epidermal lens cells facilitate the absorptance of diffuse light? AB - Many understory plants rely on diffuse light for photosynthesis because direct light is usually scattered by upper canopy layers before it strikes the forest floor. There is a considerable gap in the literature concerning the interaction of direct and diffuse light with leaves. Some understory plants have well developed lens-shaped epidermal cells, which have long been thought to increase the absorption of diffuse light. To assess the role of epidermal cell shape in capturing direct vs. diffuse light, we measured leaf reflectance and transmittance with an integrating sphere system using leaves with flat (Begonia erythrophylla, Citrus reticulata, and Ficus benjamina) and lens-shaped epidermal cells (B. bowerae, Colocasia esculenta, and Impatiens velvetea). In all species examined, more light was absorbed when leaves were irradiated with direct as opposed to diffuse light. When leaves were irradiated with diffuse light, more light was transmitted and more was reflected in both leaf types, resulting in absorptance values 2-3% lower than in leaves irradiated with direct light. These data suggest that lens-shaped epidermal cells do not aid the capture of diffuse light. Palisade and mesophyll cell anatomy and leaf thickness appear to have more influence in the capture and absorption of light than does epidermal cell shape. PMID- 21636476 TI - Differences in mechanical and structural properties of surface and aerial petioles of the aquatic plant Nymphaea odorata subsp. tuberosa (Nymphaeaceae). AB - Lily pads (Nymphaea odorata) exhibit heterophylly where a single plant may have leaves that are submerged, floating, or above (aerial) the surface of the water. Lily pads are placed in a unique situation because each leaf form is exposed to a distinctly different set of mechanical demands. While surface petioles may be loaded in tension under conditions of wind or waves, aerial petioles are loaded in compression because they must support the weight of the lamina. Using standard techniques, we compared the mechanical and morphological properties of both surface and aerial leaf petioles. Structural stiffness (EI) and the second moment of area (I) were higher in aerial petioles, although we detected no differences in other mechanical values (elastic modulus [E], extension ratio, and breaking strength). Morphologically, aerial petioles had a thicker rind, with increased collenchyma tissue and sclereid cell frequency. Aerial petioles also had a larger cross-sectional area and were more elliptical. Thus, subtle changes in the distribution of materials, rather than differences in their makeup, differentiate petiole forms. We suggest that the growth of aerial petioles may be an adaptive response to shading, allowing aerial leaves to rise above a crowded water surface. PMID- 21636477 TI - Morphology of Hydatellaceae, an anomalous aquatic family recently recognized as an early-divergent angiosperm lineage. AB - The family Hydatellaceae was recently reassigned to the early-divergent angiosperm order Nymphaeales rather than the monocot order Poales. This dramatic taxonomic adjustment allows comparison with other early-divergent angiosperms, both extant and extinct. Hydatellaceae possess some monocot-like features that could represent adaptations to an aquatic habit. Ecophysiological parallels can also be drawn from fossil taxa that are known from small achene-like diaspores, as in Hydatellaceae. Reproductive units of Hydatellaceae consist of perianthlike bracts enclosing several pistils and/or stamens. In species with bisexual reproductive units, a single unit resembles an "inside-out" flower, in which stamens are surrounded by carpels that are initiated centrifugally. Furthermore, involucre development in Trithuria submersa, with delayed growth of second whorl bracts, resembles similar delayed development of the second perianth whorl in Cabomba. Several hypotheses on the homologies of reproductive units in Hydatellaceae are explored. Currently, the most plausible interpretation is that each reproductive unit represents an aggregation of reduced unisexual apetalous flowers, which are thus very different from flowers of Nymphaeales. Each pistil in Hydatellaceae is morphologically and developmentally consistent with a solitary ascidiate carpel. However, ascidiate carpel development, consistent with placement in Nymphaeales, is closely similar to pseudomonomerous pistil development as in Poaes. PMID- 21636478 TI - Comparative infructescence morphology in Altingia (Altingiaceae) and discordance between morphological and molecular phylogenies. AB - Altingia (Altingiaceae) is a tropical to subtropical Asian genus of lowland trees for which 5-15 species have been recognized. Morphological diversity, particularly of the mature infructescence, has been poorly known, especially for species with relatively localized and narrow distributions, and our understanding of Altingia has lagged behind that of its close temperate relative Liquidambar (sweet gum). In this contribution, mature infructescence structure, at the levels of anatomy, morphology, and micromorphology, and some distinctive inflorescence features, are described for five recognized species of Altingia, some for the first time. In the phylogenetic framework of both morphology and molecules, characters of Altingia contrast with those of Liquidambar and suggest that character evolution within Altingiaceae is at least partly related to geographic and climatic distribution. Differences in rates of evolution and morphological convergence suggest complex patterns of diversification in Altingiaceae at several different phylogenetic levels: (1) at the deep nodes, characters of the stem lineage fossil Microaltingia persist into crown group Altingiaceae, morphological stasis; (2) at the generic level, convergence within both Liquidambar and Altingia toward their respective habitats; (3) at the infrageneric level, morphological divergence in species diversification within Altingia, in response to diverse habitats of the eastern Asian subtropics; and (4) within the intercontinental disjunct species pair L. orientalis-L. styraciflua, morphological stasis. PMID- 21636479 TI - Cell wall degradation and modification during programmed cell death in lace plant, Aponogeton madagascariensis (Aponogetonaceae). AB - An unusual form of leaf morphogenesis occurs in the aquatic, lace plant, Aponogeton madagascariensis (Aponogetonaceae). Early in development, discrete patches of cells undergo programmed cell death (PCD) and form perforations during leaf expansion. In addition to the protoplasts, walls of the dying cells are degraded during PCD. The cuticle of the perforation site is eroded first, followed by dissolution of cell wall matrix components, so that walls appear as loose fibrillar networks as perforations form. Gel diffusion assays of wall degrading enzyme activity indicated that pectinases are active throughout leaf development, while cellulase activity was restricted to early stages of perforation formation. Alcian blue staining showed that degrading walls remain rich in pectin, and immunolocalization of pectin epitopes indicated that the proportions of esterified and de-esterifed pectins do not change significantly. Walls of perforation border cells are modified by suberin deposition late in development, and reactive oxygen species, thought to have a role in polymerization of phenolic suberin monomers, are present at the same stage. This timing suggests that suberization may limit the spread of PCD and provide an apoplastic barrier against microbial invasion but does not initiate PCD. PMID- 21636480 TI - Plant biomechanical strategies in response to frequent disturbance: uprooting of Phyllostachys nidularia (Poaceae) growing on landslide-prone slopes in Sichuan, China. AB - Bamboo is considered useful for controlling landslides, but we observed numerous shallow-slope failures in forests of big node bamboo (Phyllostachys nidularia) in Sichuan, China. Therefore, we inventoried landslide occurrence and vegetation type along one valley. To quantify bamboo root anchorage, we performed uprooting tests and measured plant morphological characteristics. Landslide occurrence was greatest at sites with bamboo and young trees. Culm failure was common because of the high length to diameter ratio (242 +/- 6). Uprooting tests showed that the maximal force to cause failure was small (1615 +/- 195 N). Uprooting force was strongly and positively regressed with a combination of the predictors lateral root number and volume (R(2) = 0.92), and root systems were highly superficial (depth = 0.15 +/- 0.12 m), contributing little to slope stability. In P. nidularia, which grows on landslide-prone slopes, surprisingly few resources have been allocated to anchorage. We suggest that this strategy puts this pioneer at an advantage on steep slopes, where it contributes little to slope stability and colonizes frequently formed gaps through vegetative regeneration. Fewer disturbances would result in subsequent secondary succession and dying back of this shade intolerant species. PMID- 21636481 TI - Hydrochory as a determinant of genetic distribution of seeds within Hibiscus moscheutos (Malvaceae) populations. AB - Seed dispersal is a major determinant of the spatial genetic structure of plant populations. In this study, we evaluated the role of distinct hydrologic regimes in determining the spatial genetic structure of the seed bank of the wetland plant Hibiscus moscheutos. We analyzed seeds in surface soil samples collected in the autumn and the following spring by determining their allozyme genotypes and estimated the pattern in seed movements during flooding. We selected study sites in nontidal and tidal wetlands with different flooding regimes. One nontidal site had no flooding, while the second nontidal site was inundated for most of the year. One tidal wetland site flooded with almost every tide, and a second tidal site was inundated at moderate frequency. Genetic makeup of the seed bank at the nonflooded site changed little between seasons. Secondary seed dispersal altered absolute allele frequencies at the other three sites, with the greatest change occurring at the two tidally influenced sites. This study demonstrates that secondary hydrochory influences the genetic composition of the seed bank and that hydrologic conditions play an important role in determining the local patterns in seed movements. PMID- 21636482 TI - Landscape structure, clonal propagation, and genetic diversity in Scandinavian populations of Arabidopsis lyrata (Brassicaceae). AB - Colonization history, landscape structure, and environmental conditions may influence patterns of neutral genetic variation because of their effects on gene flow and reproductive mode. We compared variation at microsatellite loci within and among 26 Arabidopsis lyrata populations in two disjunct areas of its distribution in northern Europe (Norway and Sweden). The two areas probably share a common colonization history but differ in size (Norwegian range markedly larger than Swedish range), landscape structure (mountains vs. coast), and habitat conditions likely to affect patterns of gene flow and opportunities for sexual reproduction. Within-population genetic diversity was not related to latitude but was higher in Sweden than in Norway. Population differentiation was stronger among Norwegian than among Swedish populations (F(ST) = 0.23 vs. F(ST) = 0.18). The frequency of clonal propagation (proportion of identical multilocus genotypes) increased with decreasing population size, was higher in Norwegian than in Swedish populations, but was not related to altitude or substrate. Differences in genetic structure are discussed in relation to population characteristics and range size in the two areas. The results demonstrate that the possibility of clonal propagation should be considered when developing strategies for sampling and analyzing data in ecological and genetic studies of this emerging model species. PMID- 21636483 TI - Introduction history and population genetics of the invasive grass Bromus tectorum (Poaceae) in Canada. AB - The invasive annual Bromus tectorum (cheatgrass) is distributed in Canada primarily south of 52 degrees N latitude in two diffuse ranges separated by the extensive coniferous forest in western Ontario. The grass was likely introduced independently to eastern and western Canada post-1880. We detected regional variation in the grass's genetic diversity using starch gel electrophoresis to analyze genetic diversity at 25 allozyme loci in 60 populations collected across Canada. The Pgm-1a & Pgm-2a multilocus genotype, which occurs in the grass's native range in Eastern Europe, is prevalent in eastern Canada but occurs at low frequency in western Canada. In contrast, the Got-4c multilocus genotype, found in the native range in Central Europe, is widespread in populations from western Canada. Overall genetic diversity of B. tectorum is much higher in eastern Canada than in the eastern U.S., while the genetic diversity in populations in western North America is similar between Canada and the U.S. The distribution of genetic diversity across Canada strongly suggests multiple introduction events. Heterozygous individuals, which are exceedingly rare in B. tectorum, were detected in three Canadian populations. Formation of novel genotypes through occasional outcrossing events could spark adaptive evolution and further range expansion across Canada of this exceedingly damaging grass. PMID- 21636484 TI - Pollen tube growth in association with a dry-type stigmatic transmitting tissue and extragynoecial compitum in the basal angiosperm Kadsura longipedunculata (Schisandraceae). AB - Spatial features of pollen tube growth and the composition of the extracellular matrix (ECM) of transmitting tissue in carpels of Kadsura longipedunculata, a member of the basal angiosperm taxon Schisandraceae, were characterized to identify features of transmitting tissue that might have been important for pollen-carpel interactions during the early history of angiosperms. In addition to growing extracellularly along epidermal cells that make up stigmatic crests of individual carpels, pollen tubes grow on abaxial carpel epidermal cells between unfused carpels along an extragynoecial compitum to subsequently enter an adjacent carpel, a feature important for enhancing seed set in apocarpous species. Histo- and immunochemical data indicated that transmitting tissue ECM is not freely flowing as previously hypothesized. Rather, the ECM is similar to that of a dry-type stigma whereby a cuticular boundary with associated esterase activity confines a matrix containing methyl-esterified homogalacturonans. The Schisandraceae joins an increasing number of basal angiosperm taxa that have a transmitting tissue ECM similar to a dry-type stigma, thereby challenging traditional views that the ancestral pollen tube pathway was similar to a wet type stigma covered with a freely flowing exudate. Dry-type stigmas are posited to provide tighter control over pollen capture, retention, and germination than wet-type stigmas. PMID- 21636485 TI - Delayed stigma receptivity in Collinsia heterophylla (Plantaginaceae): genetic variation and adaptive significance in relation to pollen competition, delayed self-pollination, and mating-system evolution. AB - To increase our knowledge about mating-system evolution, we need to understand the relationship between specific floral traits and mating system. Species of Collinsia (Plantaginaceae) vary extensively in mating system; this variation is associated with variation in floral morphology and development and with the timing of self-pollination. Counterintuitively, large-flowered, more outcrossing species tend to have delayed stigma receptivity, reducing the amount of time that the stigma is receptive to cross-pollination before autonomous self-pollination. To understand how the timing of stigma receptivity is related to mating-system evolution, we studied in detail the timing of both stigma receptivity and self pollination (anther-stigma contact) in two greenhouse-grown populations of large flowered Collinsia heterophylla. Crosses on emasculated flowers at different stages of floral development always produced seeds, suggesting that cross fertilization can be effected by pollen arriving prior to physiological receptivity. Phenotypic and genetic variation within populations in the timing of stigma receptivity and anther-stigma contact was substantial, although slightly less for the contact. Despite strong interspecific and interpopulation correlations, we did not find an among-genet phenotypic correlation between the traits. This indicates that each trait may respond independently to selection, and the trait association may be the result of correlational selection. PMID- 21636486 TI - Gender-specific inbreeding depression in a gynodioecious plant, Geranium maculatum (Geraniaceae). AB - In gynodioecious species, females coexist with hermaphrodites in natural populations even though hermaphrodites attract more pollinators, are capable of reproducing through pollen, and can self-fertilize. This study tests the hypothesis that inbreeding depression helps to maintain females in natural populations. It also examines whether gender lineages that differ in selfing rates might experience different levels of inbreeding depression. Female and hermaphroditic lineages of the gynodioecious species Geranium maculatum were used in self, sib-cross and outcross experiments to examine inbreeding depression levels and to determine whether these levels differ between hermaphroditic and female lineages. Six fitness correlates were measured in the greenhouse and compared among pollination types and between genders. Severe inbreeding depression was found for both individual fitness traits and cumulative fitness in early life history stages. Inbreeding depression levels were slightly higher in hermaphroditic than in female lineages, but this difference was not statistically significant. Because females are unable to self-pollinate and are less likely to experience inbreeding than hermaphrodites under natural conditions, these results suggest that severe inbreeding depression could confer a selective advantage for females that could help to maintain females in natural populations. PMID- 21636487 TI - Patterns of polyploid evolution in Greek marsh orchids (Dactylorhiza; Orchidaceae) as revealed by allozymes, AFLPs, and plastid DNA data. AB - Polyploidy is common in higher plants, and speciation in polyploid complexes is usually the result of reticulate evolution. We examined variation in nuclear AFLP fingerprints, nuclear isozymes, and hypervariable plastid DNA loci to describe speciation patterns and species relationships in the Dactylorhiza incarnata/maculata polyploid complex (marsh orchids; Orchidaceae) in Greece. Several endemic taxa with restricted distribution have been described from this area, and to propose meaningful conservation priorities, detailed relationships need to be known. We identified four independently derived allopolyploid lineages, which is a pattern poorly correlated with prevailing taxonomy. Three lineages were composed of populations restricted to small areas and may be of recent origins from extant parental lineages. One lineage with wide distribution in northern Greece was characterized by several unique plastid haplotypes that were phylogenetically related and evidently older. The D. incarnata/maculata polyploid complex in Greece has high levels of genetic diversity at the polyploid level. This diversity has accumulated over a long time and may include genetic variants originating from now extinct parental populations. Our data also indicate that the Balkans may have constituted an important refuge from which northern European Dactylorhiza were recruited after the Weichselian ice age. PMID- 21636488 TI - AFLP and DNA sequence variation in an Andean domesticate, pepino (Solanum muricatum, Solanaceae): implications for evolution and domestication. AB - The pepino (Solanum muricatum) is a vegetatively propagated, domesticated native of the Andes, where it grows with wild relatives. We used AFLPs and a 1-kb sequence of the 3-methylcrotonyl-CoA carboxylase gene to study variation of 27 accessions of S. muricatum and 35 collections of 10 species of wild relatives (Solanum section Basarthrum). A total of 298 AFLP fragments and 29 DNA sequence haplotypes were detected. Cluster and principal coordinate analyses and other genetic parameters estimated from both types of markers, show that S. muricatum is closely related to the species from one of the series (Caripensia) of section Basarthrum and that >90% of the variation of the cultigen is also represented in that series. Pepino is highly diverse, either because it is not monophyletic or it has been subjected to regular introgression with wild species, or both. Although a continuous distribution of the genetic variation occurred within the cultivated species, three genetic clusters were recognized. Cluster 1 is mostly centered in Ecuador, cluster 2 in Ecuador and Peru, and cluster 3 in Colombia and Ecuador. Cluster 3 also includes all modern cultivars studied. These results and other evidence suggest that northern Ecuador/southern Colombia is the main center of pepino diversity and the center of origin. The high genetic variation of this cultigen indicates that domestication does not always produce a genetic bottleneck. PMID- 21636489 TI - Inflorescence diversification in the "finger millet clade" (Chloridoideae, Poaceae): a comparison of molecular phylogeny and developmental morphology. AB - Within the Poaceae, inflorescence diversification and its bearing on phylogeny and evolution are exceedingly complex. We used phylogenetic information of the "finger millet clade," a group of grasses with digitate inflorescences, to study the inflorescence diversification. This clade appears monophyletic in the morphological and molecular phylogenetic analyses. Three well-supported clades are shown in our cpDNA-derived phylogeny, with clades I and III consisting of species of Chloris and Microchloa, respectively, and clade II including species of Cynodon, Dactyloctenium, and Eleusine. Variation appears at different times throughout development. Changes involving primordium number and arrangement occur very early, changes involving duration of primordium activity occur much later. Characters derived from the comparison of developmental sequences were optimized onto the most parsimonious tree. The developmental characters were congruent with the molecular phylogeny. Two developmental characters may not be homologous in the Chloris subclade and the Cynodon subclade. PMID- 21636490 TI - Arbuscular mycorrhizal symbionts in Botrychium (Ophioglossaceae). AB - Many plant species are characterized by a life cycle with a long-lived, subterranean phase that is completely dependent on mycorrhizal fungal symbionts for fixed carbon. This type of life cycle is both phylogenetically and ecologically widespread and is found in diverse vascular plant lineages from the tropics to subalpine meadows. Here we report on the molecular identities of the arbuscular mycorrhizal fungi associated with the autotrophic and underground mycoheterotrophic life cycle phases of the ferns Botrychium crenulatum and B. lanceolatum. We show that the Glomus taxa found in the mycoheterotrophic life cycle phases of B. crenulatum and B. lanceolatum are also found in conspecific and heterospecific photosynthetic neighboring plants. From our DNA sequence data, we infer carbon flow from photosynthetic plants to mycoheterotrophic plants through shared glomalean fungal networks. Finally, our phylogenetic analyses identify a major Glomus clade that forms associations with mycoheterotrophic life cycle stages of B. crenulatum and B. lanceolatum. PMID- 21636491 TI - Relationships of cucumbers and melons unraveled: molecular phylogenetics of Cucumis and related genera (Benincaseae, Cucurbitaceae). AB - Cucumis (Cucurbitaceae) comprises 33 species of annuals and perennials with a major native center of diversity in tropical and southern Africa. The genus includes some economically important and widely grown vegetables such as cucumbers and melons. Monophyly of the genus has been disputed in previous studies, but with only limited sampling. Relationships within Cucumis are thus poorly understood; moreover, the validity of the closely related genera has not been thoroughly tested. The present study was undertaken to test the monophyly of Cucumis and several closely related genera, to test sectional circumscriptions within Cucumis, and to understand the biogeographical history of the genus. We sequenced the nuclear ITS and plastid trnS-trnG regions for 40 ingroup and three outgroup taxa, representing all recognized subgenera and sections. Parsimony, maximum likelihood, and Bayesian analyses found Cucumella, Oreosyce, Mukia, Myrmecosicyos, and Dicaelospermum nested within Cucumis. The clades recovered within the Cucumis complex in some instances represent the first phylogenetically derived hypothesis of relationships, whereas others correspond to previous subgeneric and sectional classifications. At least four introductions from Africa to Asia, as well as one reintroduction to Africa, are suggested within the Cucumis complex. Cucumis sativus (cucumber) is strongly supported as sister to the eastern Asian C. hystrix, whereas C. melo (melon) is strongly supported as sister to C. sagittatus in southern Africa. PMID- 21636492 TI - Foraging behavior affects pollen removal and deposition in Impatiens capensis (Balsaminaceae). AB - Flowers of most plant species are visited by a variety of animals. Some of these visitors are effective pollinators while others remove resources without transferring pollen. Studies comparing the effectiveness of different visitors as pollinators often compare taxa without considering variation in behavior within a taxon. Wilson and Thomson (Ecology 72: 1503-1507, 1991) documented the effects of honey bees and bumble bees on the pollination dynamics of Impatiens capensis. They found that pollen-collecting honey bees removed large numbers of pollen grains from anthers but deposited little of it on stigmas; bumble bees, which sought nectar, removed less pollen but deposited more of it on stigmas. It is unclear whether the low pollen transfer efficiencies of honey bees are explained by their morphology or by their pollen-collecting behavior. We repeated the work of Wilson and Thomson at a site where honey bees were foraging for nectar, not pollen. We measured the quantity of pollen remaining in anthers, the number of pollen grains deposited on stigmas, and seed production after single visits by honey bees and bumble bees. The differences between the taxa disappeared when they were foraging in a similar manner. Our results clearly demonstrate the importance of foraging behavior on the pollination effectiveness of floral visitors. PMID- 21636493 TI - Differentiation and growth traits associated with acrotony in the apple tree (Malus xdomestica, Rosaceae). AB - Branching is a key factor in the evolutionary diversification of plants and is a main criterion for plant architecture analysis. Among descriptive features, acrotony is defined as increased vigor of the vegetative proleptic branches (from dormant buds), from the proximal to the distal part of the parent growth unit. I hypothesized that acrotony could be extended to other, usually poorly described, architectural traits. The study was conducted on two architecturally contrasting apple (Malus *domestica) cultivars, 'Pitchounette' and 'Chantecler.' The proportion and size of various offspring entities were assessed according to their position along the shoot for 2 years after parent shoot growth. Acrotony was characterized by two inverse phenomena: an acropetal decrease in the proportion of latent buds and of laterals that aborted, and an acropetal increase in the proportion of reproductive laterals among growing laterals. Distally located reproductive laterals had more spur leaves and flowers and higher fruit set and flowered earlier than reproductive laterals lower on the parent annual shoot. The results suggest that the length-based criterion used for acrotony should be integrated into a general conceptual framework in which the organogenetic potential of the axillary meristem increases from the proximal to the distal part of the annual shoot, leading to greater branching density, larger offspring, and a greater propensity for flower bud formation over consecutive years. PMID- 21636494 TI - Giant cuticular pores in Eidothea zoexylocarya (Proteaceae) leaves. AB - Ubiquitous, large diameter pores have not previously been adequately demonstrated to occur in leaf cuticles. Here we show conclusively that such structures occur in Eidothea zoexylocarya, a rainforest tree species of Proteaceae restricted to the Australian Wet Tropics. The pores are abundant, large-diameter apertures (~1 MUm), that extend perpendicularly most of the way through the cuticle from the inside. They occur on both sides of the leaf, but are absent from the cuticle associated with stomatal complexes on the abaxial side. No such pores were found in any other species, including the only other species of Eidothea, E. hardeniana from New South Wales, and other species that have previously been purported to possess cuticular pores. To determine whether these pores made the cuticles more leaky to water vapor, we measured astomatous cuticular conductances to water vapor for E. zoexylocarya and seven other Proteaceae species of the Wet Tropics. Cuticular conductance for E. zoexylocarya was relatively low, indicating that the prominent pores do not increase conductance. The function of the pores is currently obscure, but the presence of both pores and an adaxial hypodermis in E. zoexylocarya but not E. hardeniana suggests evolution in response to greater environmental stresses in the tropics. PMID- 21636495 TI - Multiclavula ichthyiformis (Fungi: Basidiomycota: Cantharellales: Clavulinaceae), a remarkable new basidiolichen from Costa Rica. AB - The new basidiolichen Multiclavula ichthyiformis Nelsen, Lucking, Umana, Trest & Will-Wolf is described from Costa Rica. The new species differs from other species of Multiclavula in having a basidiocarp with tomentose stipe and flattened lamina with nonamphigenous hymenium. Molecular sequence data (ITS) confirmed its placement within Multiclavula in the Clavulinaceae (Cantharellales, Agaricomycetes). The new lichen was discovered in a Central American paramo remnant, illustrating the importance of biotic inventories of fungi and lichens to increase our knowledge of the diversity of these groups in endangered tropical ecosystems. The new species was found as part of the TICOLICHEN project in Costa Rica. PMID- 21636496 TI - Leaf traits and leaf life spans of two xeric-adapted palmettos. AB - Plants of nutrient-poor, arid environments often have leaf traits that include small size, sclerophylly, long life span, low nutrient concentration, and low photosynthetic rate. Hence, the success of two large-leaved palmettos in peninsular Florida's seasonally xeric, nutrient-impoverished uplands seems anomalous, given that their leaves are orders of magnitude larger than the leaves of sympatric species. An examination of a 16-yr data set of leaf traits and leaf life spans across four vegetative associations differing in available light showed that Serenoa repens and Sabal etonia had low rates of leaf production coupled with long leaf life spans reaching 3.5 yr in heavily shaded plants. The adaptation of these palmettos to xeric, nutrient-poor habitats has generated dwarf statures, diminished leaf sizes and numbers, increased leaf life spans, and reduced rates of leaf production relative to other palms and congeners of more mesic sites. Leaf and petiole size, plant leaf canopy area, and leaf life span increased in both palmettos with decreasing available light, helping to compensate for reduced photosynthetic rates under shaded conditions and for the high leaf construction costs of the large, thick palmetto leaves. Large leaf size in these palmettos, likely due to phylogenetic conservatism, is compensated by other leaf traits (e.g., heavily cutinized epidermises, thick laminas) that increase survival in seasonally xeric, nutrient-impoverished environments. PMID- 21636497 TI - Mycorrhizal benefit in two low arctic herbs increases with increasing temperature. AB - Climate change may influence the relationship between arctic plants and their symbiotic mycorrhizal fungi. The benefit of the symbiosis for the host plant affects vegetation succession and may be a key parameter in predicting vegetation responses to warming. We investigated the mycorrhizal benefit in the low arctic perennial herbs Potentilla crantzii and Ranunculus acris in symbiosis with the arbuscular mycorrhizal fungus Glomus claroideum. Temperature response in the mycorrhiza-mediated acquisition of nitrogen (N) and phosphorus (P), growth, and photosynthetic nutrient-use efficiency were determined. Near the average natural soil temperature (12 degrees C), mycorrhiza did not improve plant nutrient capture but significantly enhanced plant P capture at 17 degrees C. Photosynthetic nitrogen-use efficiency was higher at 17 degrees C than at 12 degrees C and was further increased by mycorrhiza at 17 degrees C. Photosynthetic phosphorus-use efficiency was not affected by temperature or mycorrhiza. Increasing the growing temperature by 5 degrees C increased the relative shoot growth rate by 15%. Mycorrhizal symbiosis did not enhance plant growth rate, but the plants gained between 20% and 90% more mycorrhiza-mediated P when grown at higher temperature. The results suggest that these low arctic species have good potential to respond positively to increasing temperatures. PMID- 21636498 TI - Evolutionary correlations of polycyclic shoot growth in Acer (Sapindaceae). AB - Two strategies have evolved in understory trees in relation to light availability: maximization of light capture and shade tolerance. In the genus Acer, light capture is favored by a suite of traits maximizing twig thickness and leaf size and minimizing the density of branching in the crown. In contrast, shade tolerance is enhanced by minimizing crown area, crown volume, and total leaf area per unit height. Maples with polycyclic shoot growth (i.e., successive flushes of shoot growth separated by a resting phase within the same vegetative period) may benefit from the prolonged growth by growing more and increasing total leaf area; thus we hypothesize that polycyclism is evolutionarily correlated with the suite of traits related to light capture. We tested this hypothesis using different phylogenetic trees to explore correlations between polycyclism and both suites of traits. Polycyclism was correlated with the suite of traits maximizing light capture, suggesting that polycyclic maples are "optimists" (i.e., they make vigorous vertical extensions in rich light) and monocyclic maples are "pessimists" (i.e., they wait in the dark understory until a gap is opened). Both strategies have been described for different floras, and interestingly, polycyclic species recruit over a wider range of environments than the monocyclic species. PMID- 21636499 TI - The molecular phylogeny of Rebutia (Cactaceae) and its allies demonstrates the influence of paleogeography on the evolution of South American mountain cacti. AB - The tropical Andes harbor a major part of the world's plant biodiversity. The montane cacti of the tribes Browningieae, Cereeae, and Trichocereeae underwent extensive radiation and thus are well suited as a model group to study the diversification of Andean plants. We reconstructed their phylogeny employing three noncoding chloroplast regions and explained it in the context of the geological history of South America. We found that the clade of cephalia-bearing cacti with naked pericarpels is centered in northeastern Brazil, whereas almost all other clades comprise Andean species. The spatial split between the clades was probably caused by the Andean uplift and the concurrent formation of intracontinental marine basins in the Tertiary. The phylogenetic reconstructions based on parsimony and Bayesian approaches do not reflect the traditional delimitation of the tribes and of the large genera. Our results suggest that Rebutia s.l. and Echinopsis s.l. are not monophyletic and that Sulcorebutia, Weingartia, and Cintia should be united into one genus. Even though this "Weingartia-complex" and the genus Gymnocalycium are similar in size and morphological diversity, Gymnocalycium has a very high molecular divergence suggesting a comparably older radiation. PMID- 21636500 TI - Inheritance of chloroplast DNA is not strictly maternal in Silene vulgaris (Caryophyllaceae): evidence from experimental crosses and natural populations. AB - Chloroplast DNA (cpDNA) is maternally inherited in the majority, but not all, of angiosperm species. The mode of inheritance of cpDNA is a critical determinant of its molecular evolution and of its population genetic structure. Here, we present the results of investigations of the inheritance of cpDNA in Silene vulgaris, a plant used in a variety of studies in which cpDNA is an important component. PCR/RFLP markers were used to compare mother and offspring cpDNA genotypes sampled from two natural populations, and mother, father, and offspring genotypes obtained from controlled greenhouse crosses. Ten of 215 offspring cpDNA genotypes studied in the controlled crosses and three of 156 offspring from natural populations did not match that of the mother, demonstrating rare nonmaternal inheritance. That the chloroplast genome is occasionally transmitted through pollen is discussed in the context of using S. vulgaris cpDNA as a marker in studies of seed dispersal and when considering the joint evolution of the chloroplast and mitochondrial genomes. PMID- 21636501 TI - Timing of self-compatibility, flower longevity, and potential for male outcross success in Leptosiphon jepsonii (Polemoniaceae). AB - When fertilization triggers flower senescence, early autonomous selfing may cause flowers to senesce before pollen has dispersed, discounting unused pollen. Selfing-induced flower senescence was examined in Leptosiphon jepsonii, a species that varies in the timing of self-compatibility. In field and greenhouse experiments, fertilization had a large effect on flower senescence; most outcrossed flowers senesced after 1 d whereas emasculated flowers lasted 2-5 d. In a comparison of inbred lines from three populations, longevity of autonomously selfed flowers of early self-compatible individuals was significantly less than that of late self-compatible individuals. In field experiments, autonomously selfed flowers were shorter-lived in a predominantly early-selfing population than in a predominantly late-selfing population. Pollen was available and viable beyond the first day of anthesis, suggesting that reductions in flower longevity caused by autonomous selfing could incur a cost to male outcross fitness. We argue that this effect is likely to be most pronounced under intermediate rates of pollinator visitation. Observed pollinator visitation rates ranged from 0.035 0.775 visits per flower per day, indicating a potential for selfing-induced flower senescence to incur pollen discounting in Leptosiphon jepsonii. PMID- 21636502 TI - Crassulacean acid metabolism photosynthesis in columnar cactus seedlings during ontogeny: the effect of light on nocturnal acidity accumulation and chlorophyll fluorescence. AB - Columnar cacti have been traditionally classified as crassulacean acid metabolism (CAM) plants, though recent research indicates some cactus seedlings employ the C(3) pathway. To verify this last result, we measured acidity fluctuations for five columnar and one globular cactus species in seedlings from 1 to 48 d old after experimental exposure to 60% and 30% full sunlight, and in adult plants in the field. Using light-response curves of chlorophyll fluorescence, we determined photosynthetic efficiency (DeltaF/Fm'), maximum electron transport rate (ETR(max)) and saturating photosynthetically active photon flux density (PPFD(sat)). All seedlings used the CAM pathway from their first day of development, and increases in nocturnal acidity depended on species, light treatment, and age. The CAM pathway was also found in adult plants. Cactus seedlings were able to acclimatize to light conditions by making photochemical adjustments, mainly by modifying the level of light at which photosystem II is saturated (PPFD(sat)). The presence of CAM in the seedlings of columnar cacti increases water-use efficiency and reduces the risk of photoinhibition. This could favor survival in the highly variable light levels characteristic of the desert environments of columnar cacti. PMID- 21636503 TI - Pollination patterns limit hybridization between two sympatric species of Narcissus (Amaryllidaceae). AB - Natural hybrids between rare and common sympatric species are commonly eradicated to avoid the potential extinction of the rare species, although there is currently no clear predictive framework to quantify this risk. As hybrids can have intrinsic value as new evolutionary pathways, further knowledge on the factors controlling hybridization is needed. In this study we evaluated the role of pollination patterns in hybridization events in two sympatric populations of Narcissus cavanillesii and N. serotinus in Portugal. Narcissus cavanillesii is a rare species, while N. serotinus is widely distributed across the Mediterranean. The hybrid, N. *perezlarae, is quite frequent in southeastern Spain but is scarce in Portugal. Reciprocal manual crossings confirmed compatibility between the two species, although hybridization was more successful when N. cavanillesii participated as female. Narcissus cavanillesii and N. serotinus only shared one pollinator, Megachile sp. (Hymenoptera), which had low visitation rates and high flower constancy. No single isolation mechanism was fully effective in preventing hybridization. Temporal displacement of flowering peaks, strong pollinator specificity, and high flower constancy in the shared pollinator all contributed to limiting hybridization in this site. In other sympatric occurrences, different phenological windows and pollination assemblages may allow greater frequency of the hybrid. PMID- 21636504 TI - Chromosome numbers, nuclear DNA content, and polyploidy in Consolea (Cactaceae), an endemic cactus of the Caribbean Islands. AB - Polyploidy, an important mechanism of plant evolution, was investigated in Consolea, an endemic Caribbean opuntioid genus represented by nine subdioecious species with very narrow distributions, including species classified as rare or threatened. Standard chromosome counting and flow cytometric analyses were used to determine chromosome numbers and ploidy of each taxon. Compared to the base number (x = 11), the mitotic and meiotic counts indicated that there are seven hexaploid (2n = 66) and two octoploid species (2n = 88); no diploids were found. Histograms of intact nuclei confirmed that all species are polyploid, with C-DNA values ranging from 4.88-9.50 pg. The variation of DNA content was significantly higher for the octoploids than for the hexaploids. Male and female sexual morphs had similar DNA content, suggesting that there are no sex chromosomes. Cytomixis between cells and microsporocytes with no chromatin were observed. This provides a mechanism whereby gametes with variable chromosome numbers are produced, influencing reproduction and promoting speciation. In conclusion, C-DNA content and chromosome number separated Consolea species into two groups, which may correspond to two phylogenetic lineages or indicate that polyploidization occurred independently, with comparable effects on C-DNA content. PMID- 21636505 TI - Aerodynamics of saccate pollen and its implications for wind pollination. AB - Pollen grains of many wind-pollinated plants contain 1-3 air-filled bladders, or sacci. Sacci are thought to help orient the pollen grain in the pollination droplet. Sacci also increase surface area of the pollen grain, yet add minimal mass, thereby increasing dispersal distance; however, this aerodynamic hypothesis has not been tested in a published study. Using scanning electron and transmission electron microscopy, mathematical modeling, and the saccate pollen of three extant conifers with structurally different pollen grains (Pinus, Falcatifolium, Dacrydium), we developed a computational model to investigate pollen flight. The model calculates terminal settling velocity based on structural characters of the pollen grain, including lengths, widths, and depths of the main body and sacci; angle of saccus rotation; and thicknesses of the saccus wall, endoreticulations, intine, and exine. The settling speeds predicted by the model were empirically validated by stroboscopic photography. This study is the first to quantitatively demonstrate the adaptive significance of sacci for the aerodynamics of wind pollination. Modeling pollen both with and without sacci indicated that sacci can reduce pollen settling speeds, thereby increasing dispersal distance, with the exception of pollen grains having robust endoreticulations and those with thick saccus walls. Furthermore, because the mathematical model is based on structural characters and error propagation methods show that the model yields valid results when sample sizes are small, the flight dynamics of fossil pollen can be investigated. Several fossils were studied, including bisaccate (Pinus, Pteruchus, Caytonanthus), monosaccate (Gothania), and nonsaccate (Monoletes) pollen types. PMID- 21636506 TI - Bud and growth-unit structure in seedlings and saplings of Nothofagus alpina (Nothofagaceae). AB - In temperate trees, axis length growth generally results from the differentiation of organs at the end of a growing season and the extension of such "preformed organs" in the next growing season. Neoformation, i.e., the simultaneous differentiation and extension of organs, has been studied for only a few species. Here we evaluated bud composition and growth unit (GU) size for seedlings and saplings of Nothofagus alpina, a valuable South American forest tree. Trunk GUs of seedlings and saplings included preformed and neoformed organs, whereas main branch GUs of saplings were entirely preformed. The size of a GU was more closely related to the number of preformed green leaves than to the number of cataphylls of its preceding bud. Proximal buds of a trunk GU had more cataphylls and less green-leaf primordia than distal buds. Individual leaf area increased from proximal to distal positions on trunk GUs. For trunk and main-branch GUs, the length/width ratio was maximum for leaves in intermediate positions. The development of large neoformed leaves at the end of the growing season could increase the photosynthetic capacity of this species in late summer, when the activity of preformed organs is likely to be decreasing. PMID- 21636507 TI - Complex distribution patterns of di-, tetra-, and hexaploid cytotypes in the European high mountain plant Senecio carniolicus (Asteraceae). AB - DNA ploidy levels were estimated using DAPI-flow cytometry of silica-dried specimens of the European mountain plant Senecio carniolicus (Asteraceae), covering its entire distribution area in the Eastern Alps (77 populations, 380 individuals) and the Carpathians (five populations, 22 individuals). A complex pattern of ploidy level variation (2x, 4x, 5x, 6x, and 7x cytotypes) was found in this species, which has been considered uniformly hexaploid. Hexaploids predominated in the Eastern Alps and was the only cytotype found in the Carpathians, while odd ploidy levels (5x, 7x) constituted a small fraction of the samples (<1.3%). Tetraploids occurred in two disjunct areas, which correspond with putative Pleistocene refugia for silicicolous alpine plants. Diploids occurred in large portions of the Alps but were absent from areas most extensively glaciated in the past. Intrapopulational cytotype mixture was detected in 22 populations-the majority involving diploids and hexaploids-with intermediate ploidy levels mostly lacking, suggesting limited gene flow and the evolution of reproductive isolation. Significant and reproducible intracytotype variation in nuclear DNA content was observed. Higher genome size in western diploids might be due to ancient introgression with the closely related S. incanus or to different evolutionary pathways in the geographically separated diploids. PMID- 21636508 TI - Expression of matK: functional and evolutionary implications. AB - Strong phylogenetic signal from matK has rendered it an invaluable gene in plant systematic and evolutionary studies at various evolutionary depths. Further, matK is proposed as the only chloroplast-encoded group II intron maturase, thus implicating MATK in chloroplast posttranscriptional processing. For a protein coding gene, matK has an unusual evolutionary mode and tempo, including relatively high substitution rates at both the nucleotide and amino acids levels. These evolutionary features have raised questions about matK function. In the current study, we examined matK RNA and protein from representative land plant species to provide insight into functional aspects of this unusual gene. We report the first evidence of a transcript for matK separate from the trnK precursor and demonstrate that a full-length MATK protein exists in five angiosperm species. We also show that matK RNA and protein levels are regulated by light and developmental stage, suggesting functional roles for this putative maturase. Specifically, matK expression increased after etiolation and decreased at 4 weeks after germination. This work provides evidence for the expression of the only putative chloroplast-encoded group II intron maturase and insight into regulation mechanisms relating to plant development and, indirectly, to photosynthesis. PMID- 21636509 TI - Origin of Hawaiian Polystichum (Dryopteridaceae) in the context of a world phylogeny. AB - A genus-wide molecular phylogeny for Polystichum and allied genera (Dryopteridaceae) was reconstructed to address the biogeographic origin and evolution of the three Hawaiian Polystichum species, all endemic there. The analysis was based on the cpDNA sequences rbcL and the trnL-F spacer from a taxonomically and geographically diverse sample. Parsimony and Bayesian phylogenetic analyses of the combined data support a monophyletic Polystichum and corroborate recent hypotheses as to membership and sequence of origin of the major groups within the genus. The Hawaiian Polystichum species are polyphyletic; two separate lineages appear to have arrived independently from the Old World. The provenance of the diploid Polystichum hillebrandii is continental eastern Asia, while the source of the polyploid lineage comprising tetraploid P. haleakalense and octoploid P. bonseyi is likely continental Asia. From our results, the origin of the Hawaiian species of Polystichum, like many Hawaiian fern genera with several species, is the result of multiple migrations to the islands, rather than single migrations yielding nearly all the local diversity as in the angiosperms. This emerging pattern provides a modern test of the premise that propagule vagility has a central role in determining pattern of evolution. PMID- 21636510 TI - Molecular phylogeny of the moonseed family (Menispermaceae): implications for morphological diversification. AB - We used the chloroplast gene ndhF to reconstruct the phylogeny of the moonseed family (Menispermaceae), a morphologically diverse and poorly known cosmopolitan family of dioecious, primarily climbing plants. This study includes a worldwide sample of DNA sequences for 88 species representing 49 of the 70 genera of all eight traditionally recognized tribes. Phylogenetic relationships were estimated, and the Shimodaira-Hasegawa test was used to compare the likelihood of alternative phylogenetic hypotheses and to evaluate the monophyly of tribes currently in use. The monospecific Indo-Malesian Tinomiscium is sister to the remaining members of the family, within which are two major clades. Within these two clades, well-supported clades correspond to four of the eight traditionally recognized tribes, while others, such as Menispermeae, are polyphyletic. Mapping of major morphological characters on the phylogeny indicates that the crescent shaped seed is derived from a straight seed, the tree habit has arisen multiple times, endosperm has been lost many times, but unicarpellate flowers evolved only once. Morphological synapomorphies for Menispermaceae include the presence of a condyle, a large embryo, and druplets. The phylogeny provides for the first time a detailed molecular-based assessment of relationships in Menispermaceae and clarifies our understanding of morphological diversification within the family. PMID- 21636511 TI - Structural attributes of the hypogeous holoparasite Hydnora triceps Drege & Meyer (Hydnoraceae). AB - The morphology of the hypogeous root holoparasite Hydnora triceps is highly reduced, and as with many holoparasites, the vegetative body is difficult to interpret. The vegetative body of H. triceps has been historically considered a "pilot root" studded with lateral appendages known as "haustorial roots." We found the vegetative body of H. triceps to consist of a rhizome with a thickened root-cap-like structure that covered a vegetative shoot apical meristem. From the apical meristem, procambial strands originated and developed into endarch collateral vascular bundles arranged radially around a pith without an interfascicular cambium. Xylem vessels had scalariform pitting and simple perforation plates. A continuous periderm without root hairs was observed. Increase in girth was attributed to cork and fascicular cambia. "Haustorial roots" or bumps on the surface of the vegetative body were exogenous, contained meristems and were the origins of vegetative branching, budding, and haustoria. The haustoria of H. triceps were cylindrical and penetrated the host root stele. Phloem and xylem elements were observed within the endophyte, and direct xylem to host-xylem contacts were observed. The arrangement of vascular tissues and xylem anatomy of H. triceps are likely plesiomorphic features in light of Hydnoraceae's placement in the Piperales. PMID- 21636512 TI - Floral sex allocation at individual and branch levels in Betula platyphylla var. japonica (Betulaceae), a tall, wind-pollinated monoecious tree species. AB - Floral sex allocation at the individual and first-order branch levels and the relation between these levels were examined in Betula platyphylla var. japonica, a wind-pollinated monoecious tree. Floral sex allocation at the individual level varied with resource availability in a pattern similar to that predicted by the Masaka and Takada model (Journal of Theoretical Biology 240: 114-125). Thus, individual trees with few reproductive resources produced only female or male inflorescences, whereas individuals with many resources rarely had a high male ratio (i.e., number of male inflorescences/total number of inflorescences). Furthermore, the number of male inflorescences tended to reach an upper limit, whereas the number of female inflorescences increased monotonically with increasing reproductive investment. The patterns of floral sex allocation at the first-order branch level were analogous to those at the individual level. Thus, each first-order branch of B. platyphylla var. japonica behaves like an individual, and the floral sex allocation of a given branch does not necessarily represent the individual tree. The effect of the individual-level floral sex ratio on branch-level floral sex allocation indicates that branch behavior is controlled by the individual. PMID- 21636513 TI - Functional diversity of carbon-gain, water-use, and leaf-allocation traits in trees of a threatened lowland dry forest in Hawaii. AB - We examined carbon-gain, water-use, and leaf-allocation traits for six tree species of a Hawaiian dry forest to better understand the functional diversity within this threatened ecosystem. Tropical dry forests are among the most endangered ecosystems on Earth, and in Hawaii, as elsewhere, declining biodiversity threatens ecosystem processes that may depend on forest functional diversity. We found broad variation among species including a two-fold difference for mean photosynthetic rate, a greater than three-fold difference for predawn water potential, and a nearly three-fold difference for leaf life span. Principal component analysis showed a clear separation of species based on carbon-gain vs. water-use related axes, and delta(13)C analysis revealed differing limitations (supply vs. demand) on carbon assimilation. The broad functional variation not only spanned traditional classifications (avoiders vs. tolerators), but also included unusual strategies (e.g., fast growth with drought tolerance). Correlations among traits, including leaf life span, leaf mass per area, and %N, followed typical global patterns, but some exceptions appeared as a result of unique life-history characteristics, such as latex-rich sap and root parasitism. Elucidating functional variation provides important information that can be used to link plant biodiversity with ecosystem processes and also facilitate the management and preservation of tropical dry forests and other threatened communities. PMID- 21636514 TI - Impact of global warming on a group of related species and their hybrids: cherry tree (Rosaceae) flowering at Mt. Takao, Japan. AB - Climate change is affecting plant phenology worldwide. Phenological responses vary among species, but it is not clear how responses differ among closely related species. We examined a 25-yr record (1981-2005) of flowering times for 97 trees, representing 17 species and hybrids of cherry (Cerasus sp. or Prunus sp.) grown at Mt. Takao, in Tokyo, Japan. The cherry trees flowered earlier over time, by an average of 5.5 d over the 25-yr study. Earlier flowering was explained largely by a 1.8 degrees C increase in February-March mean monthly temperatures. Most species and hybrids flowered 3-5 d earlier for each 1 degrees C increase in temperature, but early-flowering taxa flowered as much as 9 d earlier for each 1 degrees C increase in temperature. Flowering durations and differences in flowering times among species were greater in warm years than in cold years. Species and individual trees also flowered longer in warm years. These results show that the flowering times of closely related species may change similarly in response to climate change, but that early-flowering species may diverge from the overall trend in a predictable way. Such changes in flowering may affect gene flow and pollination as the length of the flowering season increases. PMID- 21636515 TI - Genetic consequences of pre-Columbian cultivation for Agave murpheyi and A. delamateri (Agavaceae). AB - Pre-Columbian farmers cultivated several species of agave in central Arizona from ca. A.D. 600-1350. Because of the longevity and primarily asexual reproduction of these species, relict agave clones remain in the landscape and provide insights into pre-Columbian agricultural practices. We analyzed variation in allozyme allele frequencies to infer genetic effects of prehistoric cultivation on Agave murpheyi and A. delamateri, specifically to estimate genetic diversity and structure, to determine whether cultivated populations descended from a single clone, and to examine regional-scale genetic variation. Agave murpheyi maintained more genetic diversity at the species and population levels than A. delamateri, and A. murpheyi populations typically included more multilocus genotypes. Relict plants from prehistoric fields reflect a more complex history than descent from a single clone; A. murpheyi populations may have included more diversity initially because bulbils (produced routinely in A. murpheyi but not A. delamateri) and possibly seed would have facilitated transport of genetically diverse planting stock. Genetic variation in both cultigens was lower than in most contemporary commercial crops but similar to that observed in modern traditional agricultural systems. PMID- 21636516 TI - Boreotropical migration explains hybridization between geographically distant lineages in the pantropical clade Sideroxyleae (Sapotaceae). AB - To determine whether the fragmented pantropical distribution of present day Sideroxyleae primarily is the result of long-distance dispersals or represents the remnants of a once continuous distribution in the northern hemisphere, the boreotropical flora, we used phylogenetic analyses of chloroplast and nuclear ribosomal DNA data, Bayesian molecular dating, and Bayesian estimation of ancestral areas. Incongruence between the two data sets was examined with a nuclear low copy gene phylogeny to discover any occurrences of reticulate evolution. The Pacific clade Nesoluma was shown to have two distinct copies of the nuclear low copy gene AAT, one from an African and one from an American ancestral lineage, indicating that it is of allopolyploid origin. We conclude that Sideroxyleae, including the ancestral lineages of Nesoluma, were part of the boreotropical flora and entered the New World via the north Atlantic land bridge. We also suggest that the distribution of extant species resulted from the cooling climate at the end of the Eocene. Sideroxylon oxyacanthum is shown not to belong in the group, but in Chrysophylloideae. A classification reflecting phylogenetic relationships, as well as new combinations for the species in Nesoluma under Sideroxylon, is presented. PMID- 21636517 TI - A generic 3D finite element model of tree anchorage integrating soil mechanics and real root system architecture. AB - Understanding the mechanism of tree anchorage in a forest is a priority because of the increase in wind storms in recent years and their projected recurrence as a consequence of global warming. To characterize anchorage mechanisms during tree uprooting, we developed a generic finite element model where real three dimensional (3D) root system architectures were represented in a 3D soil. The model was used to simulate tree overturning during wind loading, and results compared with real data from two poplar species (Populus trichocarpa and P. deltoides). These trees were winched sideways until failure, and uprooting force and root architecture measured. The uprooting force was higher for P. deltoides than P. trichocarpa, probably due to its higher root volume and thicker lateral roots. Results from the model showed that soil type influences failure modes. In frictional soils, e.g., sandy soils, plastic failure of the soil occurred mainly on the windward side of the tree. In cohesive soils, e.g., clay soils, a more symmetrical slip surface was formed. Root systems were more resistant to uprooting in cohesive soil than in frictional soil. Applications of this generic model include virtual uprooting experiments, where each component of anchorage can be tested individually. PMID- 21636518 TI - Molecular phylogeny suggests a single origin of insect symbiosis in the Pucciniomycetes with support for some relationships within the genus Septobasidium. AB - In the Pucciniomycetes, a class of fungi that includes the plant pathogenic rust fungi, insect parasitism is restricted to a single family, the Septobasidiaceae. The Septobasidiaceae form a variety of symbioses with scale insects and have remained largely unstudied since the 1930s. Transitions between plant and animal parasitism and between mutualism and parasitism cannot be fully addressed in the Basidiomycota without a clear phylogenetic hypothesis for the Septobasidiales. Here, molecular phylogenetic methods were applied to understand the origin of scale insect parasitism, test the monophyly of the order Septobasidiales, and evaluate the infrageneric concepts in the largest genus of scale insect parasites, Septobasidium. DNA sequence data from rRNA genes were used to infer higher-level relationships within the Pucciniomycetes, and data from translation elongation factor 1-alpha (tef1) were added for phylogenetic inference within the Septobasidiaceae. Data from tef1 revealed different intron arrangements within Septobasidium, but the molecule did not provide much additional phylogenetically informative data. Likelihood-model-based phylogenetic analyses of 44 Pucciniomycotina taxa provided moderate support for a single origin of insect parasitism. Within the Septobasidiaceae, there was little or no support for a monophyletic Septobasidium, and well-resolved subclades of Septobasidium species contradict previous morphological delimitations of groups within the genus. PMID- 21636519 TI - Polyploidy and self-fertilization in flowering plants. AB - Mating systems directly control the transmission of genes across generations, and understanding the diversity and distribution of mating systems is central to understanding the evolution of any group of organisms. This basic idea has been the motivation for many studies that have explored the relationships between plant mating systems and other biological and/or ecological phenomena, including a variety of floral and environmental characteristics, conspecific and pollinator densities, growth form, parity, and genetic architecture. In addition to these examples, a potentially important but poorly understood association is the relationship between plant mating systems and genome duplication, i.e., polyploidy. It is widely held that polyploid plants self-fertilize more than their diploid relatives, yet a formal analysis of this pattern does not exist. Data from 235 species of flowering plants were used to analyze the association between self-fertilization and ploidy. Phylogenetically independent contrasts and cross-species analyses both lend support to the hypothesis that polyploids self fertilize more than diploids. Because polyploidy and self-fertilization are so common among angiosperms, these results contribute not only to our understanding of the relationship between mating systems and polyploidy in particular, but more generally, to our understanding of the evolution of flowering plants. PMID- 21636520 TI - Seed morphology of modern and fossil Ampelocissus (Vitaceae) and implications for phytogeography. AB - Seeds are useful in distinguishing among extant genera of Vitaceae and provide a good basis for interpretation of fossil remains in reconstructing the evolutionary and phytogeographic history of this putatively basal Rosid family. Seeds of Ampelocissus s.l. including Pterisanthes and Nothocissus are distinguished from those of all other vitaceous genera by long, parallel ventral infolds and a centrally positioned oval chalazal scar. Principal component analysis facilitates recognition of four Ampelocissus s.l. seed morphotypes differentiated by dorsiventral thickness, width of ventral infolds, chalazal depth, and degree of dorsal surface rugosity. While these intergrade, their end member morphologies are distinctive and coincide well with inflorescence morphology, extant geographic distribution, and ecology. Seven fossil morphospecies are recognized. Ampelocissus parvisemina sp. n. (Paleocene of North Dakota; Eocene of Oregon) and A. auriforma Manchester (Eocene of Oregon) resemble extant Central American species; A. bravoi Berry (Eocene of Peru) is similar to one group of Old World extant species; and A. parachandleri sp. n. (Eocene of Oregon) and the three European fossil species A. chandleri (Kirchheimer) comb. n., A. lobatum (Chandler) comb. n., and A. wildei sp. n. (Eocene to Miocene) resemble another group of extant Old World Ampelocissus. All these fossils occur outside the present geographic range of the genus, reflecting warmer climates and former intercontinental links. PMID- 21636521 TI - Phylogeny of the tribe Aveneae (Pooideae, Poaceae) inferred from plastid trnT-F and nuclear ITS sequences. AB - New insights into evolutionary trends in the economically important oat tribe (Aveneae) are presented. Plastid trnT-F and nuclear ribosomal ITS sequences were used to reconstruct the phylogeny of the Aveneae-Poeae-Seslerieae complex (Pooideae, Poaceae) through Bayesian- and maximum parsimony-based analyses, separately and in combination. The plastid data identified a strongly supported core Aveneae lineage that separated from other former Aveneae and Poeae groups. Koeleriinae, Aveninae, and Agrostidinae emerged as the main groups of this core Aveneae, which also included other minor subgroups with uncertain relationships and a few former Poeae members. Several former Aveneae representatives were also placed in independent sublineages in Poeae. Seslerieae resolved as close allies of Poeae or Aveneae in the plastid and nuclear topologies, respectively. Because of the intermingling of some Aveneae and Seslerieae lineages in Poeae and vice versa, we propose to expand Poeae to include all the aforementioned lineages. This best reflects our current understanding of the phylogeny of these important temperate grasses and sheds light on their evolutionary history. PMID- 21636522 TI - Variation in vegetative and flowering phenology in a forest herb caused by environmental heterogeneity. AB - Timing of seasonal plant development can affect biotic interactions and plant fitness. Phenology is governed largely by temperature and may therefore be affected by global climate warming, making this an important area of research. Several factors in addition to temperature may cause differences in phenology. We studied the influence of local environment, plant size, and reproductive effort on shoot emergence and flowering time of 290 individuals of Actaea spicata (Ranunculaceae), distributed among 25 plots in four populations. We used multiple regression and structural equation models (SEM) to study causal relationships. Among plots, soil temperature and canopy cover explained 63% of the variation in shoot emergence. Soil temperature, slope, and canopy cover together explained 83% of the variation in flowering time. Within plots, small plants on steep south facing slopes with high soil potassium concentrations emerged earlier in the year. Plants emerging earlier flowered earlier, but no environmental factors affected flowering time directly. We found no effects of reproductive effort. Our results support the view that flowering time of temperate forest herbs is constrained by several environmental factors acting indirectly through effects on shoot emergence time. PMID- 21636523 TI - Vivipary in coastal cacti: a potential reproductive strategy in halophytic environments. AB - Vivipary, the germination of seeds within the fruit prior to abscission from the maternal plant, is an important event in plants. Two main types of vivipary are known in vascular plants: true vivipary and pseudovivipary. In crop grasses, pseudovivipary is an undesirable character as it results in lower yields. To date, vivipary in the Cactaceae has been reported for less than 20 species, most of which are cultivated. Here, we report viviparous (cryptoviviparous-a subcategory of true vivipary) cacti in nature in members of the tribes Cacteae and Pachycereeae (subfamily Cactoideae). We present four species inhabiting coastal plains in areas subject to periodic flooding, namely, Ferocactus herrerae, Stenocereus alamosensis, S. thurberi, and Pachycereus schottii. These species from localities in northwestern Mexico had viviparous fruits and offspring in different stages of development. A potential trend in the data indicates that the overall proportion of viviparous plants is higher in coastal flooding areas relative to halophytic, nonflooding areas. In our view, vivipary is a reproductive strategy that has evolved to provide a more efficient mechanism favoring germination and new avenues for survival by contributing to population maintenance and short-distance dispersal on halophytic substrates. PMID- 21636524 TI - The spatial tuning of achromatic and chromatic vision in budgerigars. AB - Birds are assumed to use half of their cones (double cones) to detect fine spatial detail while their other half (single cones) is used for color vision. However, the spatial resolution of the color pathway in birds has never been studied. We determined the spatial contrast sensitivity to achromatic and isoluminant red-green and blue-green color gratings in budgerigars (Melopsittacus undulatus). Contrast sensitivity to achromatic gratings has band-pass characteristics while that for red-green and blue-green gratings has low-pass properties. Maximum sensitivity is lower to blue-green than to red-green gratings and the acuity for both color gratings is less than half (ca. 4.5 cycles/degree) of that for achromatic gratings (ca. 10 cycles/degree). This suggests that achromatic vision in birds, as in humans and bees, is tuned for detecting fine detail while chromatic vision is tuned for viewing larger fields. Similar to humans, blue-sensitive cones contribute little to spatial vision. Moreover, budgerigars detected gratings having both achromatic and chromatic contrasts more reliably at high spatial frequencies than gratings with either of these contrasts, suggesting that the single and double cone pathways are incompletely separated. The study demonstrates the importance of the spatial dimension of color vision; fine patterns remain unresolved even if they present large color contrasts. PMID- 21636525 TI - Detection of staphylococcal cassette chromosome mec type XI carrying highly divergent mecA, mecI, mecR1, blaZ, and ccr genes in human clinical isolates of clonal complex 130 methicillin-resistant Staphylococcus aureus. AB - Methicillin resistance in staphylococci is mediated by penicillin binding protein 2a (PBP 2a), encoded by mecA on mobile staphylococcal cassette chromosome mec (SCCmec) elements. In this study, two clonal complex 130 (CC130) methicillin resistant Staphylococcus aureus (MRSA) isolates from patients in Irish hospitals were identified that were phenotypically PBP 2a positive but lacked mecA by conventional PCR and by DNA microarray screening. The isolates were identified as methicillin-susceptible S. aureus using the GeneXpert real-time PCR assay. Whole genome sequencing of one isolate (M10/0061) revealed a 30-kb SCCmec element encoding a class E mec complex with highly divergent blaZ-mecA-mecR1-mecI, a type 8 cassette chromosome recombinase (ccr) complex consisting of ccrA1-ccrB3, an arsenic resistance operon, and flanking direct repeats (DRs). The SCCmec element was almost identical to that of SCCmec type XI (SCCmec XI) identified by the Sanger Institute in sequence type 425 bovine MRSA strain LGA251 listed on the website of the International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements. The open reading frames (ORFs) identified within SCCmec XI of M10/0061 exhibited 21 to 93% amino acid identity to ORFs in GenBank. A third DR was identified ca. 3 kb downstream of SCCmec XI, indicating the presence of a possible SCC remnant. SCCmec XI was also identified in the second CC130 MRSA isolate by PCR and sequencing. The CC130 MRSA isolates may be of animal origin as previously reported CC130 S. aureus strains were predominantly from bovine sources. The highly divergent nature of SCCmec XI relative to other SCCmec elements indicates that it may have originated in another taxon. PMID- 21636526 TI - Retrotransposition of marked SVA elements by human L1s in cultured cells. AB - Human retrotransposons generate structural variation and genomic diversity through ongoing retrotransposition and non-allelic homologous recombination. Cell culture retrotransposition assays have provided great insight into the genomic impact of retrotransposons, in particular, LINE-1(L1) and Alu elements; however, no such assay exists for the youngest active human retrotransposon, SINE-VNTR-Alu (SVA). Here we report the development of an SVA cell culture retrotransposition assay. We marked several SVAs with either neomycin or EGFP retrotransposition indicator cassettes. Engineered SVAs retrotranspose using L1 proteins supplemented in trans in multiple cell lines, including U2OS osteosarcoma cells where SVA retrotransposition is equal to that of an engineered L1. Engineered SVAs retrotranspose at 1-54 times the frequency of a marked pseudogene in HeLa HA cells. Furthermore, our data suggest a variable requirement for L1 ORF1p for SVA retrotransposition. Recovered engineered SVA insertions display all the hallmarks of LINE-1 retrotransposition and some contain 5' and 3' transductions, which are common for genomic SVAs. Of particular interest is the fact that four out of five insertions recovered from one SVA are full-length, with the 5' end of these insertions beginning within 5 nt of the CMV promoter transcriptional start site. This assay demonstrates that SVA elements are indeed mobilized in trans by L1. Previously intractable questions regarding SVA biology can now be addressed. PMID- 21636527 TI - Wild-type HTT modulates the enzymatic activity of the neuronal palmitoyl transferase HIP14. AB - Huntington disease (HD) is caused by polyglutamine expansion in the huntingtin (HTT) protein. Huntingtin-interacting protein 14 (HIP14), one of 23 DHHC domain containing palmitoyl acyl transferases (PATs), binds to HTT and robustly palmitoylates HTT at cysteine 214. Mutant HTT exhibits reduced palmitoylation and interaction with HIP14, contributing to the neuronal dysfunction associated with HD. In this study, we confirmed that, among 23 DHHC PATs, HIP14 and its homolog DHHC-13 (HIP14L) are the two major PATs that palmitoylate HTT. Wild-type HTT, in addition to serving as a palmitoylation substrate, also modulates the palmitoylation of HIP14 itself. In vivo, HIP14 palmitoylation is decreased in the brains of mice lacking one HTT allele (hdh+/-) and is further reduced in mouse cortical neurons treated with HTT antisense oligos (HTT-ASO) that knockdown HTT expression by ~95%. Previously, it has been shown that palmitoylation of DHHC proteins may affect their enzymatic activity. Indeed, palmitoylation of SNAP25 by HIP14 is potentiated in vitro in the presence of wild-type HTT. This influence of HTT on HIP14 activity is lost in the presence of CAG expansion. Furthermore, in both brains of hdh+/- mice and neurons treated with HTT-ASO, we observe a significant reduction in palmitoylation of endogenous SNAP25 and GluR1, synaptic proteins that are substrates of HIP14, suggesting wild-type HTT also influences HIP14 enzymatic activity in vivo. This study describes an important biochemical function for wild-type HTT modulation of HIP14 palmitoylation and its enzymatic activity. PMID- 21636528 TI - DNMT1 deficiency triggers mismatch repair defects in human cells through depletion of repair protein levels in a process involving the DNA damage response. AB - DNA methyltransferase 1 (DNMT1) maintains methylation at CpG dinucleotides, important for transcriptional silencing at many loci. It is also implicated in stabilizing repeat sequences: DNMT1 deficiency causes microsatellite instability in mouse embryonic stem cells, but it is unclear how this occurs, how repeats lacking CpG become unstable and whether the effect is confined to stem cells. To address these questions, we transfected hTERT-immortalized normal human fibroblasts (hTERT-1604) with a short hairpin RNA construct targeting DNMT1 and isolated stable integrants with different levels of protein. DNMT1 expression levels agreed well with methylation levels at imprinted genes. Knockdown cells showed two key characteristics of mismatch repair (MMR) deficiency, namely resistance to the drug 6-thioguanine and up to 10-fold elevated mutation rates at a CA(17) microsatellite reporter, but had limited viability. The likely cause of MMR defects is a matching drop in steady-state protein levels for key repair components in DNMT1 knockdown cells, affecting both the MutLalpha and MutSalpha complexes. This indirect effect on MMR proteins was also seen using a different targeting method in HT29 colon cancer cells and did not involve transcriptional silencing of the respective genes. Decreased levels of MMR components follow activation of the DNA damage response and blocking this response, and in particular poly(ADP-ribose) polymerase (PARP) overactivation, rescues cell viability in DNMT1-depleted cells. These results offer an explanation for how and why unmethylated microsatellite repeats can be destabilized in cells with decreased DNMT1 levels and uncover a novel and important role for PARP in this process. PMID- 21636529 TI - Abdominal surgery inhibits circulating acyl ghrelin and ghrelin-O-acyltransferase levels in rats: role of the somatostatin receptor subtype 2. AB - Clinical studies are evaluating the efficacy of synthetic ghrelin agonists in postoperative ileus management. However, the control of ghrelin secretion under conditions of postoperative gastric ileus is largely unknown. Peripheral somatostatin inhibits ghrelin secretion in animals and humans. We investigated the time course of ghrelin changes postsurgery in fasted rats and whether somatostatin receptor subtype 2 (sst(2)) signaling is involved. Abdominal surgery (laparotomy and 1-min cecal palpation) induced a rapid and long-lasting decrease in plasma acyl ghrelin levels as shown by the 64, 67, and 59% reduction at 0.5, 2, and 5 h postsurgery, respectively, compared with sham (anesthesia alone for 10 min, P < 0.05). Levels were partly recovered at 7 h and fully restored at 24 h. The percentage of acyl ghrelin reduction was significantly higher than that of desacyl ghrelin at 2 h postsurgery and not at any other time point. This was associated with a 48 and 23% decrease in gastric and plasma ghrelin-O acyltransferase protein concentrations, respectively (P < 0.001). Ghrelin positive cells in the oxyntic mucosa expressed sst(2a) receptor and the sst(2) agonist S-346-011 inhibited fasting acyl ghrelin levels by 64 and 77% at 0.5 and 2 h, respectively. The sst(2) antagonist S-406-028 prevented the abdominal surgery-induced decreased circulating acyl ghrelin but not the delayed gastric emptying assessed 0.5 h postinjection. These data show that activation of sst(2) receptor located on gastric X/A-like cells plays a key role in the rapid inhibition of circulating acyl ghrelin induced by abdominal surgery while not being primarily involved in the early phase of postoperative gastric ileus. PMID- 21636530 TI - Expression, localization, and functional role for synaptotagmins in pancreatic acinar cells. AB - Secretagogue-induced changes in intracellular Ca(2+) play a pivotal role in secretion in pancreatic acini yet the molecules that respond to Ca(2+) are uncertain. Zymogen granule (ZG) exocytosis is regulated by soluble N ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexes. In nerve and endocrine cells, Ca(2+)-stimulated exocytosis is regulated by the SNARE associated family of proteins termed synaptotagmins. This study examined a potential role for synaptotagmins in acinar secretion. RT-PCR revealed that synaptotagmin isoforms 1, 3, 6, and 7 are present in isolated acini. Immunoblotting and immunofluorescence using three different antibodies demonstrated synaptotagmin 1 immunoreactivity in apical cytoplasm and ZG fractions of acini, where it colocalized with vesicle-associated membrane protein 2. Synaptotagmin 3 immunoreactivity was detected in membrane fractions and colocalized with an endolysosomal marker. A potential functional role for synaptotagmin 1 in secretion was indicated by results that introduction of synaptotagmin 1 C2AB domain into permeabilized acini inhibited Ca(2+)-dependent exocytosis by 35%. In contrast, constructs of synaptotagmin 3 had no effect. Confirmation of these findings was achieved by incubating intact acini with an antibody specific to the intraluminal domain of synaptotagmin 1, which is externalized following exocytosis. Externalized synaptotagmin 1 was detected exclusively along the apical membrane. Treatment with CCK-8 (100 pM, 5 min) enhanced immunoreactivity by fourfold, demonstrating that synaptotagmin is inserted into the apical membrane during ZG fusion. Collectively, these data indicate that acini express synaptotagmin 1 and support that it plays a functional role in secretion whereas synaptotagmin 3 has an alternative role in endolysosomal membrane trafficking. PMID- 21636531 TI - Oxidative stress-induced posttranslational modification of TRPV1 expressed in esophageal epithelial cells. AB - Human esophageal epithelium is continuously exposed to physical stimuli or to gastric acid that sometimes causes inflammation of the mucosa. Transient receptor potential vanilloid 1 (TRPV1) is a nociceptive, Ca(2+)-selective ion channel activated by capsaicin, heat, and protons. It has been reported that activation of TRPV1 expressed in esophageal mucosa is involved in gastroesophageal reflux disease (GERD) or in nonerosive GERD symptoms. In this study, we examined the expression and function of TRPV1 in the human esophageal epithelial cell line Het1A, focusing in particular on the role of oxidative stress. Interleukin-8 (IL 8) secreted by Het1A cells upon stimulation by capsaicin or acid with/without 4 hydroxy-2-nonenal (HNE) was measured by ELISA. Following capsaicin stimulation, the intracellular production of reactive oxygen species (ROS) was determined using a redox-sensitive fluorogenic probe, and ROS- and HNE-modified proteins were determined by Western blotting using biotinylated cysteine and anti-HNE antibody, respectively. HNE modification of TRPV1 proteins was further investigated by immunoprecipitation after treatment with synthetic HNE. Capsaicin and acid induced IL-8 production in Het1A cells, and this production was diminished by antagonists of TRPV1. Capsaicin also significantly increased the production of intracellular ROS and ROS- or HNE-modified proteins in Het1A cells. Moreover, IL-8 production in capsaicin-stimulated Het1A cells was enhanced by synthetic HNE treatment. Immunoprecipitation studies revealed that TRPV1 was modified by HNE in synthetic HNE-stimulated Het1A cells. We concluded that TRPV1 functions in chemokine production in esophageal epithelial cells, and this function may be regulated by ROS via posttranslational modification of TRPV1. PMID- 21636532 TI - Deoxycholic acid causes DNA damage while inducing apoptotic resistance through NF kappaB activation in benign Barrett's epithelial cells. AB - Gastroesophageal reflux is associated with adenocarcinoma in Barrett's esophagus, but the incidence of this tumor is rising, despite widespread use of acid suppressing medications. This suggests that refluxed material other than acid might contribute to carcinogenesis. We looked for potentially carcinogenetic effects of two bile acids, deoxycholic acid (DCA) and ursodeoxycholic acid (UDCA), on Barrett's epithelial cells in vitro and in vivo. We exposed Barrett's (BAR-T) cells to DCA or UDCA and studied the generation of reactive oxygen/nitrogen species (ROS/RNS); expression of phosphorylated H2AX (a marker of DNA damage), phosphorylated IkBalpha, and phosphorylated p65 (activated NF-kappaB pathway proteins); and apoptosis. During endoscopy in patients, we took biopsy specimens of Barrett's mucosa before and after esophageal perfusion with DCA or UDCA and assessed DNA damage and NF-kappaB activation. Exposure to DCA, but not UDCA, resulted in ROS/RNS production, DNA damage, and NF-kappaB activation but did not increase the rate of apoptosis in BAR-T cells. Pretreatment with N-acetyl l-cysteine (a ROS scavenger) prevented DNA damage after DCA exposure, and DCA did induce apoptosis in cells treated with NF-kappaB inhibitors (BAY 11-7085 or AdIkappaB superrepressor). DNA damage and NF-kappaB activation were detected in biopsy specimens of Barrett's mucosa taken after esophageal perfusion with DCA, but not UDCA. These data show that, in Barrett's epithelial cells, DCA induces ROS/RNS production, which causes genotoxic injury, and simultaneously induces activation of the NF-kappaB pathway, which enables cells with DNA damage to resist apoptosis. We have demonstrated molecular mechanisms whereby bile reflux might contribute to carcinogenesis in Barrett's esophagus. PMID- 21636533 TI - Branched-chain amino acids increase arterial blood ammonia in spite of enhanced intrinsic muscle ammonia metabolism in patients with cirrhosis and healthy subjects. AB - Branched-chain amino acids (BCAA) are used in attempts to reduce blood ammonia in patients with cirrhosis and intermittent hepatic encephalopathy based on the hypothesis that BCAA stimulate muscle ammonia detoxification. We studied the effects of an oral dose of BCAA on the skeletal muscle metabolism of ammonia and amino acids in 14 patients with cirrhosis and in 7 healthy subjects by combining [(13)N]ammonia positron emission tomography (PET) of the thigh muscle with measurements of blood flow and arteriovenous (A-V) concentrations of ammonia and amino acids. PET was used to measure the metabolism of blood-supplied ammonia and the A-V measurements were used to measure the total ammonia metabolism across the thigh muscle. After intake of BCAA, blood ammonia increased more than 30% in both groups of subjects (both P < 0.05). Muscle clearance of blood-supplied ammonia (PET) was unaffected (P = 0.75), but the metabolic removal rate (PET) increased significantly because of increased blood ammonia in both groups (all P < 0.05). The total ammonia clearance across the leg muscle (A-V) increased by more than 50% in both groups, and the flux (A-V) of ammonia increased by more than 45% (all P < 0.05). BCAA intake led to a massive glutamine release from the muscle (cirrhotic patients, P < 0.05; healthy subjects, P = 0.12). In conclusion, BCAA enhanced the intrinsic muscle metabolism of ammonia but not the metabolism of blood-supplied ammonia in both the patients with cirrhosis and in the healthy subjects. PMID- 21636535 TI - Accountable care organizations versus accountable care: is there a difference? PMID- 21636536 TI - Head and neck cancers. PMID- 21636537 TI - Controversies in the locoregional management of head and neck cancer. AB - Head and neck cancer (HNC) is a heterogeneous combination of various sites and types of disease. This manuscript elaborates on 3 important and current issues: the emerging role of human papilloma virus (HPV) in oropharyngeal cancer (OPC), current considerations in systemic therapy for advanced disease, and evolving treatment of the neck. Exogenous carcinogens, most notably tobacco, have classically been implicated in the development of HNC. A large increase in the incidence of OPC has occurred in the past few decades, predominantly in nontobacco users, and is caused by HPV. This disease is unique in many respects and presents an opportunity for novel therapeutic approaches. Because the prognosis for HPV-related HNC is better, regardless of whether surgery or radiation is used as the primary therapy, the reduction of treatment-related morbidity has assumed increasing importance and provides unique opportunities and challenges for de-escalation of therapies. Radiotherapy (RT) and concurrent cisplatin is the most commonly used nonsurgical platform for locally advanced disease. New data suggest that viable alternatives exist to the typical 3 cycles of bolus high-dose cisplatin. The role of RT and concurrent taxanes remains less understood. Similarly, the value of integrating epidermal growth factor inhibition and concurrent chemoradiation is under continuing investigation. The use of PET scanning is changing the traditional use of adjuvant neck dissection after RT or chemoradiation. Recent data support the use of surgery in the presence of a positive posttreatment PET, and observation in the setting of a negative posttreatment scan. PMID- 21636538 TI - The epidemic of HPV-associated oropharyngeal cancer is here: is it time to change our treatment paradigms? AB - Although relatively uncommon, oropharyngeal cancers are increasing in incidence despite declining prevalence of smoking and in direct opposition to a decreasing incidence of all other head and neck cancers. An epidemic of human papillomavirus (HPV)--associated oropharyngeal cancers seems to account for these incidence trends. Important demographic, behavioral, and prognostic characteristics define this unique population. Changes in prevention, diagnosis, evaluation, staging, and treatment are needed. This article summarizes the epidemiology and clinical behavior of HPV-associated oropharyngeal cancer and discusses evolving/potential paradigms of treatment. However, data are currently insufficient to change treatment paradigms for HPV-associated oropharyngeal cancer outside of a closely monitored clinical trial. PMID- 21636539 TI - Current recommendations for systemic therapy of recurrent and/or metastatic head and neck squamous cell cancer. AB - For palliation of patients with recurrent and/or metastatic head and neck squamous cell cancer (R/M HNSCC), the major classes of commonly used cytotoxic chemotherapeutic agents are platinum agents (cisplatin, carboplatin), taxanes (paclitaxel, docetaxel), and antimetabolic agents (methotrexate, 5-fluorouracil). Cetuximab, a monoclonal antibody directed against the extracellular domain of the epidermal growth factor receptor, also shows modest activity against R/M HNSCC. Because the overall management of patients with R/M HNSCC often involves multidisciplinary input, this review focuses on data that help guide decision making in scenarios in which palliative chemotherapy is planned. Avenues for ongoing research are also presented. PMID- 21636540 TI - Direct and differential effects of stem cell factor on the neovascularization activity of endothelial progenitor cells. AB - AIMS: Previous studies on the role of stem cell factor (SCF) in endothelial progenitor cell (EPC)-mediated neovascularization have focused on the EPC mobilization and homing process. However, the direct effects of SCF on neovascularization activity of EPCs have not been characterized. We sought to determine whether SCF regulates the neovascularization ability of EPCs by comparing its roles in mature endothelial cells. METHODS AND RESULTS: In vitro and in vivo assays revealed that SCF substantially increased the neovascularization activity of human EPCs through the c-Kit receptor. Notably, the SCF-induced increase in neovascularization activity was substantially greater in EPCs than that in human umbilical vein endothelial cells (HUVECs). SCF-induced phosphorylation of c-Kit and downstream signalling molecules was consistently found to be more potent and longer-lasting in EPCs than in HUVECs. This high responsiveness of EPCs to SCF was explained by the finding that the cell-surface expression of c-Kit is far higher in EPCs than in HUVECs. A c-Kit promoter assay revealed that the increased expression of c-Kit in EPCs could be attributed to the greater expression of stem cell leukaemia, LIM-only 2, and GATA-binding protein 2. CONCLUSION: In addition to its documented role in the mobilization and recruitment of EPCs, our findings show that SCF directly enhances the neovascularization activity of EPCs. Furthermore, the present study provides further evidence that EPCs exhibit differentially greater responsiveness to hypoxia-inducible cytokines, including SCF, than mature endothelial cells, suggesting that EPCs in ischaemic tissues function differently from mature endothelial cells, although they exhibit very similar phenotypes. PMID- 21636541 TI - Nuclear morphometry identifies a distinct aggressive cellular phenotype in cutaneous squamous cell carcinoma. AB - By identifying aggressive cutaneous squamous cell carcinoma (cSCC) in patients who are at high risk for recurrences or second primaries after resection, intensive surveillance and therapy may decrease morbidity and mortality. We investigated the role of nuclear morphometry (karyometry) in differentiating between aggressive and nonaggressive cSCC. We retrospectively analyzed cSCC lesions from 40 male patients. Twenty-two patients had evidence of aggressive cSCC (local/regional recurrence or a second primary cSCC), and 18 patients were identified with similar ages and sites of disease as control patients with nonaggressive cSCC (no evidence of recurrence, metastasis, or second primary). We carried out karyometric analysis to identify nuclear features that discriminate between aggressive and nonaggressive cSCC nuclei. We used statistically significant differences (Kruskal-Wallis test, P < 0.0001) to compose a quantitative aggressive classification score (proportion of aggressive nuclei from 0% to 100%). For comparisons, we used Fisher's exact test or Student's t test. The mean age was 79 +/- 7 years for aggressive cSCC and 80 +/- 9 years for nonaggressive cSCC (P = 0.66). We analyzed a mean of 96 nuclei in each group. The mean classification score for aggressive cSCC was significantly higher (69% +/- 6%) than for nonaggressive cSCC (28% +/- 5%, P = 0.00002). Overall, the classification score accurately categorized 80% of our patients (P = 0.0004). In most patients, karyometry differentiated between aggressive and nonaggressive cSCC. We found that classification scores, which provide information on individual lesions, could be used for risk stratification. PMID- 21636542 TI - Altered histology provides a positive clinical signal in the bronchial epithelium. AB - The history of lung cancer chemoprevention trials has been uniformly disappointing in that the large phase III studies showed no effect or harm in actively smoking participants, and smaller phase II studies have also been negative. In this issue of the journal (beginning on page 793), Keith and colleagues report their randomized, placebo-controlled trial of the oral prostacyclin analogue iloprost, the first trial to show an improvement in bronchial histology (i.e., regression), which occurred in former, but not current, smokers with sputum atypia. This Perspective discusses the strength of the clinical signal provided by this observation and its implications for further drug development. PMID- 21636543 TI - Cotargeting cyclin D1 starts a new chapter in lung cancer prevention and therapy. AB - Lung cancer has limited effective therapy and no effective prevention. Cytotoxic chemotherapy has not improved when combined with the epidermal growth factor receptor (EGFR) inhibitor erlotinib (standard lung cancer therapy) or with the rexinoid bexarotene. Combining erlotinib and bexarotene, however, to cotarget cyclin D1 via the retinoid X receptor and EGFR was active preclinically in KRAS driven lung cancer cells derived from transgenic mice and in two clinical studies in lung cancer (including wild-type EGFR tumors, with or without KRAS mutations), as reported in this issue of the journal by Dragnev and colleagues (beginning on page 818). These results, along with closely related clinical results of the BATTLE program, support the promise of this cotargeting approach for lung cancer prevention and therapy and of cyclin D1 as a predictive, personalizing marker for it. PMID- 21636544 TI - Biomarkers in exploring the frontiers of diagnosis, prognosis, and therapy of Barrett's esophagus. AB - Barrett's esophagus (BE) is the principal risk factor for esophageal adenocarcinoma. BE patients currently undergo periodic endoscopic surveillance with tissue sampling and histopathologic assessment for dysplasia. They frequently are prescribed proton pump inhibitors to pharmacologically suppress gastric acid that is the cause of BE. These standard endoscopic and pharmacologic approaches for managing BE are crude at best. Identification of novel tissue biomarkers within BE may allow for more accurate endoscopic risk stratification and provide potential targets for chemoprevention. PMID- 21636545 TI - Cancer interception. AB - A common perception is that cancer risk reduction is passive, such as not smoking. However, advances in the understanding of cancer biology and in cancer treatment modalities suggest that it is now timely to consider anew cancer risk reduction by active, including pharmacologic, approaches. Risk avoidance approaches are certainly important, but other approaches are important as well, as exemplified by the irony that most new lung cancers occur in former smokers, or current avoiders. Cancer interception is the active way of combating cancer and carcinogenesis at earlier and earlier stages. A great challenge is to educate people that the development of cancers, like heart disease, typically takes years and accordingly can potentially be intercepted with risk-reducing agents in the same way that advanced cancers can be treated with drugs or that cardiovascular disease can be intercepted with antihypertensive and other risk-reducing drugs. The cancer biology behind cancer interception is increasingly solid. For example, hedgehog pathway studies of mutations in the patched homolog 1 (PTCH1) gene, which constitutively activates Smoothened (SMO), led to development of an oral SMO inhibitor active in advanced basal cell carcinoma and which, in very high risk Gorlin syndrome patients (germ line PTCH1 mutation), is nearly completely clinically effective in intercepting basal cell neoplasia. Also, the oral immunomodulator lenalidomide, first found to be active in advanced, relapsed multiple myeloma, was highly effective in intercepting the precursor stage, high risk smoldering multiple myeloma from progressing. These are but two exciting, recent examples of the many advances in cancer research that have created an optimal time to discover and implement cancer interception. The multifaceted roles of telomere maintenance in both fueling advanced cancers and, at early stages, keeping them at bay, also highlight how the growing knowledge of cancer biology opens avenues for cancer interception. Emerging molecular techniques, including next-generation sequencing platforms, that account for a large part of the remarkable recent advances in cancer biology are now being applied to interception of premalignancy. Keeping the medical community and public at large informed about possibilities for actively intercepting cancer will be important for gaining acceptance of this increasingly powerful approach to lessening the cancer burden. PMID- 21636546 TI - Oral iloprost improves endobronchial dysplasia in former smokers. AB - There are no established chemopreventive agents for lung cancer, the leading cause of cancer death in the United States. Prostacyclin levels are low in lung cancer and supplementation prevents lung cancer in preclinical models. We carried out a multicenter double-blind, randomized, phase II placebo-controlled trial of oral iloprost in current or former smokers with sputum cytologic atypia or endobronchial dysplasia. Bronchoscopy was performed at study entry and after completion of six months of therapy. Within each subject, the results were calculated by using the average score of all biopsies (Avg), the worst biopsy score (Max), and the dysplasia index (DI). Change in Avg was the primary end point, evaluated in all subjects, as well as in current and former smokers. The accrual goal of 152 subjects was reached and 125 completed both bronchoscopies (60/75 iloprost, 65/77 placebo). Treatment groups were well matched for age, tobacco exposure, and baseline histology. Baseline histology was significantly worse for current smokers (Avg 3.0) than former smokers (Avg 2.1). When compared with placebo, former smokers receiving oral iloprost exhibited a significantly greater improvement in Avg (0.41 units better, P = 0.010), in Max (1.10 units better, P = 0.002), and in DI (12.45%, P = 0.006). No histologic improvement occurred in current smokers. Oral iloprost significantly improves endobronchial histology in former smokers and deserves further study to determine if it can prevent the development of lung cancer. PMID- 21636547 TI - Characterizing the impact of smoking and lung cancer on the airway transcriptome using RNA-Seq. AB - Cigarette smoke creates a molecular field of injury in epithelial cells that line the respiratory tract. We hypothesized that transcriptome sequencing (RNA-Seq) will enhance our understanding of the field of molecular injury in response to tobacco smoke exposure and lung cancer pathogenesis by identifying gene expression differences not interrogated or accurately measured by microarrays. We sequenced the high-molecular-weight fraction of total RNA (>200 nt) from pooled bronchial airway epithelial cell brushings (n = 3 patients per pool) obtained during bronchoscopy from healthy never smoker (NS) and current smoker (S) volunteers and smokers with (C) and without (NC) lung cancer undergoing lung nodule resection surgery. RNA-Seq libraries were prepared using 2 distinct approaches, one capable of capturing non-polyadenylated RNA (the prototype NuGEN Ovation RNA-Seq protocol) and the other designed to measure only polyadenylated RNA (the standard Illumina mRNA-Seq protocol) followed by sequencing generating approximately 29 million 36 nt reads per pool and approximately 22 million 75 nt paired-end reads per pool, respectively. The NuGEN protocol captured additional transcripts not detected by the Illumina protocol at the expense of reduced coverage of polyadenylated transcripts, while longer read lengths and a paired end sequencing strategy significantly improved the number of reads that could be aligned to the genome. The aligned reads derived from the two complementary protocols were used to define the compendium of genes expressed in the airway epithelium (n = 20,573 genes). Pathways related to the metabolism of xenobiotics by cytochrome P450, retinol metabolism, and oxidoreductase activity were enriched among genes differentially expressed in smokers, whereas chemokine signaling pathways, cytokine-cytokine receptor interactions, and cell adhesion molecules were enriched among genes differentially expressed in smokers with lung cancer. There was a significant correlation between the RNA-Seq gene expression data and Affymetrix microarray data generated from the same samples (P < 0.001); however, the RNA-Seq data detected additional smoking- and cancer-related transcripts whose expression was were either not interrogated by or was not found to be significantly altered when using microarrays, including smoking-related changes in the inflammatory genes S100A8 and S100A9 and cancer-related changes in MUC5AC and secretoglobin (SCGB3A1). Quantitative real-time PCR confirmed differential expression of select genes and non-coding RNAs within individual samples. These results demonstrate that transcriptome sequencing has the potential to provide new insights into the biology of the airway field of injury associated with smoking and lung cancer. The measurement of both coding and non-coding transcripts by RNA-Seq has the potential to help elucidate mechanisms of response to tobacco smoke and to identify additional biomarkers of lung cancer risk and novel targets for chemoprevention. PMID- 21636549 TI - Evaluation of difluoromethylornithine for the chemoprevention of Barrett's esophagus and mucosal dysplasia. AB - Patients with Barrett's esophagus (BE) and dysplasia are candidates for chemopreventive strategies to reduce cancer risk. We determined the effects of difluoromethylornithine (DMFO) on mucosal polyamines, gene expression, and histopathology in BE. Ten patients with BE and low-grade dysplasia participated in a single-arm study of DFMO (0.5 g/m(2)/d) given continuously for 6 months. Esophagoscopy with biopsies was conducted at baseline, 3, 6, and 12 months. Dysplasia was graded by a gastrointestinal pathologist. Audiology was assessed (at baseline and at 6 months). Mucosal polyamines were measured by high performance liquid chromatography. Microarray-based gene expression was analyzed using a cDNA two-color chip. DFMO suppressed levels of the polyamines putrescine (P = 0.02) and spermidine (P = 0.02) and the spermidine/spermine ratio (P < 0.01) in dysplastic BE (6 months vs. baseline) that persisted at 6 months following drug cessation. Among the top 25 modulated genes, we found those regulating p53 mediated cell signaling (RPL11), cell-cycle regulation (cyclin E2), and cell adhesion and invasion (Plexin1). DFMO downregulated Kruppel-like factor 5 (KLF5), a transcription factor promoting cell proliferation, and suppressed RFC5 whose protein interacts with proliferating cell nuclear antigen. Histopathology showed regression of dysplasia (n = 1), stable disease (n = 8), and progression to high grade dysplasia (n = 1). Polyamines were suppressed in the responder to a greater extent than in stable cases. DFMO was well tolerated, and one patient had subclinical, unilateral ototoxicity. DFMO suppressed mucosal polyamines and modulated genes that may be mechanistically related to its chemopreventive effect. Further study of DFMO for the chemoprevention of esophageal cancer in BE patients is warranted. PMID- 21636548 TI - Bexarotene plus erlotinib suppress lung carcinogenesis independent of KRAS mutations in two clinical trials and transgenic models. AB - The rexinoid bexarotene represses cyclin D1 by causing its proteasomal degradation. The epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) erlotinib represses cyclin D1 via different mechanisms. We conducted a preclinical study and 2 clinical/translational trials (a window-of opportunity and phase II) of bexarotene plus erlotinib. The combination repressed growth and cyclin D1 expression in cyclin-E- and KRAS/p53-driven transgenic lung cancer cells. The window-of-opportunity trial in early-stage non-small-cell lung cancer (NSCLC) patients (10 evaluable), including cases with KRAS mutations, repressed cyclin D1 (in tumor biopsies and buccal swabs) and induced necrosis and inflammatory responses. The phase II trial in heavily pretreated, advanced NSCLC patients (40 evaluable; a median of two prior relapses per patient (range, 0-5); 21% with prior EGFR-inhibitor therapy) produced three major clinical responses in patients with prolonged progression-free survival (583-, 665-, and 1,460-plus days). Median overall survival was 22 weeks. Hypertriglyceridemia was associated with an increased median overall survival (P = 0.001). Early PET (positron emission tomographic) response did not reliably predict clinical response. The combination was generally well tolerated, with toxicities similar to those of the single agents. In conclusion, bexarotene plus erlotinib was active in KRAS-driven lung cancer cells, was biologically active in early-stage mutant KRAS NSCLC, and was clinically active in advanced, chemotherapy-refractory mutant KRAS tumors in this study and previous trials. Additional lung cancer therapy or prevention trials with this oral regimen are warranted. PMID- 21636550 TI - Clonal structure of carcinogen-induced intestinal tumors in mice. AB - Previous studies have shown that intestinal tumors from Apc(Min)(/+) (Min) mice and familial adenomatous polyposis (FAP) patients are often polyclonal. We sought to determine whether polyclonality is unique to tumors arising from hereditary predispositions or, instead, is a common feature of intestinal tumorigenesis in other pathways to tumorigenesis. Ethylnitrosourea-induced intestinal tumors from mice wild type at the Apc locus and chimeric for the Rosa26 lineage marker were analyzed. Many were overtly polyclonal, being composed of a mixture of Rosa26(+) and Rosa26(-) neoplastic cells. Statistical analyses revealed that polyclonality could be explained by interactions between two initiated clones separated by a very short distance. The frequency of overtly polyclonal tumors and the range of interactions estimated in this model are similar to those observed when analyzing familial tumors from Min mice. Thus, polyclonality does not depend on the familial pathway to tumorigenesis. Interactions between two initiated clones might provide a selective advantage during the early stages of intestinal tumorigenesis. PMID- 21636552 TI - BRAF activation induces transformation and then senescence in human neural stem cells: a pilocytic astrocytoma model. AB - PURPOSE: BRAF is frequently activated by gene fusion or point mutation in pilocytic astrocytoma, the most common pediatric brain tumor. We investigated the functional effect of constitutive BRAF activation in normal human neural stem and progenitor cells to determine its role in tumor induction in the brain. EXPERIMENTAL DESIGN: The constitutively active BRAF(V600E) allele was introduced into human neurospheres, and its effects on MAPK (mitogen-activated protein kinase) signaling, proliferation, soft agarose colony formation, stem cell phenotype, and induction of cellular senescence were assayed. Immunohistochemistry was used to examine p16(INK4a) levels in pilocytic astrocytoma. RESULTS: BRAF(V600E) expression initially strongly promoted colony formation but did not lead to significantly increased proliferation. BRAF(V600E) expressing cells subsequently stopped proliferating and induced markers of oncogene-induced senescence including acidic beta-galactosidase, PAI-1, and p16(INK4a) whereas controls did not. Onset of senescence was associated with decreased expression of neural stem cell markers including SOX2. Primary pilocytic astrocytoma cultures also showed induction of acidic beta-galactosidase activity. Immunohistochemical examination of 66 pilocytic astrocytomas revealed p16(INK4a) immunoreactivity in the majority of cases, but patients with tumors negative for p16(INK4a) had significantly shorter overall survival. CONCLUSIONS: BRAF activation in human neural stem and progenitor cells initially promotes clonogenic growth in soft agarose, suggesting partial cellular transformation, but oncogene-induced senescence subsequently limits proliferation. Induction of senescence by BRAF may help explain the low-grade pathobiology of pilocytic astrocytoma, whereas worse clinical outcomes associated with tumors lacking p16(INK4a) expression could reflect failure to induce senescence or an escape from oncogene-induced senescence. PMID- 21636553 TI - Role of CXCR3 ligands in IL-7/IL-7R alpha-Fc-mediated antitumor activity in lung cancer. AB - PURPOSE: We evaluated the utility of chimeric gammac homeostatic cytokine, IL 7/IL-7Ralpha-Fc, to restore host APC (antigen presenting cell) and T cell activities in lung cancer. EXPERIMENTAL DESIGN: Utilizing murine lung cancer models we determined the antitumor efficacy of IL-7/IL-7Ralpha-Fc. APC, T cell, cytokine analyses, neutralization of CXCL9, CXCL10, and IFNgamma were carried out to evaluate the mechanistic differences in the antitumor activity of IL-7/IL 7Ralpha-Fc in comparison to controls. RESULTS: IL-7/IL-7Ralpha-Fc administration inhibited tumor growth and increased survival in lung cancer. Accompanying the tumor growth inhibition were increases in APC and T cell activities. In comparison to controls, IL-7/IL-7Ralpha-Fc treatment of tumor bearing mice led to increased: (i) levels of CXCL9, CXCL10, IFNgamma, IL-12 but reduced IL-10 and TGFbeta, (ii) tumor macrophage infiltrates characteristic of M1 phenotype with increased IL-12, iNOS but reduced IL-10 and arginase, (iii) frequencies of T and NK cells, (iv) T cell activation markers CXCR3, CD69 and CD127(low), (v) effector memory T cells, and (vi) T cell cytolytic activity against parental tumor cells. IL-7/IL-7Ralpha-Fc treatment abrogated the tumor induced reduction in splenic functional APC activity to T responder cells. The CXCR3 ligands played an important role in IL-7/IL-7Ralpha-Fc-mediated antitumor activity. Neutralization of CXCL9, CXCL10, or IFNgamma reduced CXCR3 expressing activated T cells infiltrating the tumor and abrogated IL-7/IL-7Ralpha-Fc-mediated tumor growth inhibition. CONCLUSIONS: Our findings show that IL-7/IL-7Ralpha-Fc promotes afferent and efferent antitumor responses in lung cancer. PMID- 21636555 TI - Infection control in UK neonatal units: the greater impact in surgical units. PMID- 21636554 TI - Genetic variations in multiple drug action pathways and survival in advanced stage non-small cell lung cancer treated with chemotherapy. AB - PURPOSE: Variations in genes related to biological activity of anticancer drugs could influence treatment responses and lung cancer prognosis. Genetic variants in four biological pathways, that is, glutathione metabolism, DNA repair, cell cycle, and epidermal growth factor receptor (EGFR), were systematically investigated to examine their association with survival in advanced stage non small cell lung cancer (NSCLC) treated with chemotherapy. EXPERIMENTAL DESIGN: A total of 894 tagging single-nucleotide polymorphisms (SNP) in 70 genes from the four pathways were genotyped and analyzed in a 1,076-patient cohort. Association with overall survival was analyzed at SNP and whole-gene levels within all patients and major chemotherapy agent combination groups. RESULTS: A poorer overall survival was observed in patients with genetic variations in GSS (glutathione pathway) and MAP3K1 (EGFR pathway; HR = 1.45; 95% CI = 1.20-1.77 and HR = 1.25; 95% CI = 1.05-1.50, respectively). In the stratified analysis on patients receiving platinum plus taxane treatment, we observed a hazardous effect on overall survival by the MAP3K1 variant (HR = 1.38; 95% CI = 1.11-1.72) and a protective effect by RAF1 (HR = 0.64; 95% CI = 0.50-0.82) in the EGFR pathway. In patients receiving platinum plus gemcitabine treatment, RAF1 and GPX5 (glutathione pathway) genetic variations showed protective effects on survival (HR = 0.54; 95% CI = 0.38-0.77; HR = 0.67; 95% CI = 0.52-0.85, respectively); in contrast, NRAS (EGFR pathway) and GPX7 (glutathione pathway) variations showed hazardous effects on overall survival (HR = 1.91; 95% CI = 1.30-2.80; HR = 1.83; 95% CI = 1.27-2.63, respectively). All genes that harbored these significant SNPs remained significant by whole-gene analysis. CONCLUSION: Common genetic variations in genes of EGFR and glutathione pathways may be associated with overall survival among patients with advanced stage NSCLC treated with platinum, taxane, and/or gemicitabine combinations. PMID- 21636556 TI - Who understands delirium? PMID- 21636557 TI - Dose response of commercially available optically stimulated luminescent detector, Al2O3:C for megavoltage photons and electrons. AB - This study examined the dose response of an optically stimulated luminescence dosemeter (OSLD) to megavoltage photon and electron beams. A nanoDotTM dosemeter was used to measure the dose response of the OSLD. Photons of 6-15 MV and electrons of 9-20 MeV were delivered by a Varian 21iX machine (Varian Medical System, Inc. Milpitas, CA, USA). The energy dependency was <1 %. For the 6-MV photons, the dose was linear until 200 cGy. The superficial dose measurements revealed photon irradiation to have an angular dependency. The nanoDotTM dosemeter has potential use as an in vivo dosimetric tool that is independent of the energy, has dose linearity and a rapid response compared with normal in vivo dosimetric tools, such as thermoluminescence detectors. However, the OSLD must be treated very carefully due to the high angular dependency of the photon beam. PMID- 21636558 TI - Development of a light-weight portable neutron survey meter. AB - A light-weight portable neutron survey meter was developed using a mixed organic gas counter for dose management at nuclear power plants and accelerator facilities. This survey meter, NSN31041, is ~2 kg in weight and W160*H250*L300 mm(3) in size, which is capable of measuring neutron ambient dose equivalent rate from thermal to 15 MeV neutrons. The neutron energy response of the survey meter is evaluated using continuous energy neutron sources of (252)Cf, (241)Am-Be, thermal neutrons generated from a graphite pile loading a (252)Cf source, concrete-moderated neutrons of (241)Am-Be source and D(2)O-moderated neutrons of (252)Cf source. The measured response data show very good agreement with neutron ambient dose equivalent within a 50 % deviation. PMID- 21636559 TI - Indoor radon measurements in the granodiorite area of Bergama (Pergamon)-Kozak, Turkey. AB - Indoor radon levels in 20 dwellings of rural areas at the Kozak-Bergama (Pergamon) granodiorite area in Turkey were measured by the alpha track etch integrated method. These dwellings were monitored for eight successive months. Results show that the radon levels varied widely in the area ranging from 11+/-1 to 727+/-11 Bq m(-3) and the geometric mean was found to be 63 Bq m(-3) with a geometric standard deviation of 2 Bq m(-3). A log-normal distribution of the radon concentration was obtained for the studied area. Estimated annual effective doses due to the indoor radon ranged from 0.27 to 18.34 mSv y(-1) with a mean value of 1.95 mSv y(-1), which is lower than the effective dose values 3-10 mSv given as the range of action levels recommended by International Commission on Radiation Protection. All dosimetric calculations were performed based on the guidance of the UNSCEAR 2000 report. PMID- 21636560 TI - Effects of simulated reflux laryngitis on laryngeal chemoreflexes in newborn lambs. AB - It has been suggested that reflux laryngitis (RL) is involved in apneas bradycardias of the newborn. The aim of the present study was to develop a unique RL model in newborn lambs to test the hypothesis that RL enhances the cardiorespiratory components of the laryngeal chemoreflexes (LCR) in the neonatal period. Gastric juice surrogate (2 ml of normal saline solution with HCl pH 2 + pepsin 300 U/ml) (RL group, n = 6) or normal saline (control group, n = 6) was repeatedly injected onto the posterior aspect of the larynx, 3 times a day for 6 consecutive days, via a retrograde catheter introduced into the cervical esophagus. Lambs instilled with gastric juice surrogate presented clinical signs of RL, as well as moderate laryngitis on histological observation. Laryngeal chemoreflexes were thereafter induced during sleep by injection of 0.5 ml of HCl (pH 2), ewe's milk, distilled water or saline into the laryngeal vestibule via a chronic, transcutaneous supraglottal catheter. Overall, RL led to a significantly greater respiratory inhibition compared with the control group during LCR, including longer apnea duration (P = 0.01), lower minimal respiratory rate (P = 0.002), and a more prominent decrease in arterial hemoglobin saturation (SpO(2)) (P = 0.03). No effects were observed on cardiac variables. In conclusion, 1) our unique neonatal ovine model presents clinical and histological characteristics of RL; and 2) the presence of RL in newborn lambs increases the respiratory inhibition observed with LCR, at times leading to severe apneas and desaturations. PMID- 21636561 TI - Increased palmitoyl-myristoyl-phosphatidylcholine in neonatal rat surfactant is lung specific and correlates with oral myristic acid supply. AB - Surfactant predominantly comprises phosphatidylcholine (PC) species, together with phosphatidylglycerols, phosphatidylinositols, neutral lipids, and surfactant proteins-A to -D. Together, dipalmitoyl-PC (PC16:0/16:0), palmitoyl-myristoyl-PC (PC16:0/14:0), and palmitoyl-palmitoleoyl-PC (PC16:0/16:1) make up 75-80% of mammalian surfactant PC, the proportions of which vary during development and in chronic lung diseases. PC16:0/14:0, which exerts specific effects on macrophage differentiation in vitro, increases in surfactant during alveolarization (at the expense of PC16:0/16:0), a prenatal event in humans but postnatal in rats. The mechanisms responsible and the significance of this reversible increase are, however, not understood. We hypothesized that, in rats, myristic acid (C14:0) enriched milk is key to lung-specific PC16:0/14:0 increases in surfactant. We found that surfactant PC16:0/14:0 in suckling rats correlates with C14:0 concentration in plasma chylomicrons and lung tissue triglycerides, and that PC16:0/14:0 fractions reflect exogenous C14:0 supply. Significantly, C14:0 was increased neither in plasma PC, nor in liver triglycerides, free fatty acids, or PC. Lauric acid was also abundant in triglycerides, but was not incorporated into surfactant PC. Comparing a C14:0-rich milk diet with a C14:0-poor carbohydrate diet revealed increased C14:0 and decreased C16:0 in plasma and lung triglycerides, respectively. PC16:0/14:0 enrichment at the expense of PC16:0/16:0 did not impair surfactant surface tension function. However, the PC profile of the alveolar macrophages from the milk-fed animals changed from PC16:0/16:0 rich to PC16:0/14:0 rich. This was accompanied by reduced reactive oxygen species production. We propose that nutritional supply with C14:0 and its lung-specific enrichment may contribute to decreased reactive oxygen species production during alveolarization. PMID- 21636562 TI - Inflammatory cells and apoptosis in respiratory and limb muscles of patients with COPD. AB - Discrepancies exist regarding the involvement of cellular inflammation and apoptosis in the muscle dysfunction of chronic obstructive pulmonary disease (COPD) patients with preserved body composition. We explored whether levels of inflammatory cells and apoptosis were increased in both respiratory and limb muscles of COPD patients without nutritional abnormalities. In the vastus lateralis, external intercostals, and diaphragms of severe and moderate COPD patients with normal body composition, and in healthy subjects, intramuscular leukocytes and macrophage levels were determined (immunohistochemistry). Muscle structure was also evaluated. In the diaphragm and vastus lateralis of severe and moderate COPD patients and controls, apoptotic nuclei were explored using the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay, electron microscopy, and caspase-3 expression. In COPD patients compared with controls, diaphragm and intercostal levels of inflammatory cells were extremely low and not significantly different. However, in the vastus lateralis of the severe patients, inflammatory cell counts, although also very low, were significantly greater. In those patients, TUNEL-positive nuclei levels were also significantly greater in diaphragms and vastus lateralis. A significant inverse relationship was found between quadriceps TUNEL-positive nuclei levels and muscle force. Ultrastructural apoptotic nuclei revealed no differences in respiratory or limb muscles between COPD patients and controls. Muscle caspase-3 expression did not differ between patients and controls. In severe COPD patients with preserved body composition, while increased apoptotic nuclei seems to be a contributor to their muscle dysfunction, cellular inflammation does not. The increased numbers of TUNEL-positive nuclei in their muscles suggest that they may also be exposed to a continuous repair/remodeling process. PMID- 21636563 TI - Lower vascular tone and larger plasma volume in Parkinson's disease with orthostatic hypotension. AB - The pathophysiology of orthostatic hypotension in Parkinson's disease (PD) is incompletely understood. The primary focus has thus far been on failure of the baroreflex, a central mediated vasoconstrictor mechanism. Here, we test the role of two other possible factors: 1) a reduced peripheral vasoconstriction (which may contribute because PD includes a generalized sympathetic denervation); and 2) an inadequate plasma volume (which may explain why plasma volume expansion can manage orthostatic hypotension in PD). We included 11 PD patients with orthostatic hypotension (PD + OH), 14 PD patients without orthostatic hypotension (PD - OH), and 15 age-matched healthy controls. Leg blood flow was examined using duplex ultrasound during 60 degrees head-up tilt. Leg vascular resistance was calculated as the arterial-venous pressure gradient divided by blood flow. In a subset of 9 PD + OH, 9 PD - OH, and 8 controls, plasma volume was determined by indicator dilution method with radiolabeled albumin ((125)I-HSA). The basal leg vascular resistance was significantly lower in PD + OH (0.7 +/- 0.3 mmHg.ml( 1).min) compared with PD - OH (1.3 +/- 0.6 mmHg.ml(-1).min, P < 0.01) and controls (1.3 +/- 0.5 mmHg.ml(-1).min, P < 0.01). Leg vascular resistance increased significantly during 60 degrees head-up tilt with no significant difference between the groups. Plasma volume was significantly larger in PD + OH (3,869 +/- 265 ml) compared with PD - OH (3,123 +/- 377 ml, P < 0.01) and controls (3,204 +/- 537 ml, P < 0.01). These results indicate that PD + OH have a lower basal leg vascular resistance in combination with a larger plasma volume compared with PD - OH and controls. Despite the increase in leg vascular resistance during 60 degrees head-up tilt, PD + OH are unable to maintain their blood pressure. PMID- 21636564 TI - Metabolic disruptions induced by reduced ambulatory activity in free-living humans. AB - Physical inactivity likely plays a role in the development of insulin resistance and obesity; however, direct evidence is minimal and mechanisms of action remain unknown. Studying metabolic outcomes that occur after transitioning from higher to lower levels of physical activity is the best tool to answer these questions. Previous studies have successfully used more extreme models of inactivity, including bed rest, or the cessation of exercise in highly trained endurance athletes, to provide novel findings. However, these models do not accurately reflect the type of inactivity experienced by a large majority of the population. Recent studies have used a more applicable model in which active (~10,000 steps/day), healthy young controls are asked to transition to an inactive lifestyle (~1,500 steps/day) for a 14-day period. The transition to inactivity resulted in reduced insulin sensitivity and increased central adiposity. This review will discuss the outcomes of these studies, their implications for the cause/effect relationship between central adiposity and insulin resistance, and provide rationale for why inactivity induces these factors. In addition, the experimental challenges of directly linking acute responses to inactivity to chronic disease will also be discussed. PMID- 21636565 TI - Angiotensin II evokes sensory long-term facilitation of the carotid body via NADPH oxidase. AB - We previously reported that reactive oxygen species generated by NADPH oxidase 2 (Nox2) induces sensory plasticity of the carotid body, manifested as a progressive increase in baseline sensory activity or sensory long-term facilitation (sLTF). ANG II, a peptide generated within the carotid body, is a potent activator of Nox2. In the present study, we tested the hypothesis that ANG II evokes sLTF of the carotid body via Nox2 activation. Experiments were performed on carotid bodies ex vivo from adult rats and mice. Sensory activity was recorded from the carotid sinus nerve. Repetitive (5 times for 30 s each at 5 min intervals), but not continuous (for 150 s), application of 60 pM ANG II evoked robust sLTF of the carotid body. ACh, ATP, substance P, and KCl, when applied repetitively, stimulated the carotid body but did not evoke sLTF. Reactive oxygen species levels increased in response to repetitive applications of ANG II, and this effect was blocked by apocynin, an inhibitor of Nox2, as well as losartan, an angiotensin type 1 (AT(1)) receptor antagonist. Losartan, apocynin, and 4-(2-aminoethyl)benzenesulfonyl fluoride prevented ANG II-induced sLTF, which was absent in mice deficient in gp91(phox), the catalytic subunit of the Nox2 complex. These results demonstrate that repetitive application of ANG II induces sLTF of the carotid body via activation of Nox2 by AT(1) receptors. PMID- 21636566 TI - Acute mountain sickness, inflammation, and permeability: new insights from a blood biomarker study. AB - The pathophysiology of acute mountain sickness (AMS) is unknown. One hypothesis is that hypoxia induces biochemical changes that disrupt the blood-brain barrier (BBB) and, subsequently, lead to the development of cerebral edema and the defining symptoms of AMS. This study explores the relationship between AMS and biomarkers thought to protect against or contribute to BBB disruption. Twenty healthy volunteers participated in a series of hypobaric hypoxia trials distinguished by pretreatment with placebo, acetazolamide (250 mg), or dexamethasone (4 mg), administered using a randomized, double-blind, placebo controlled, crossover design. Each trial included peripheral blood sampling and AMS assessment before (-15 and 0 h) and during (0.5, 4, and 9 h) a 10-h hypoxic exposure (barometric pressure = 425 mmHg). Anti-inflammatory and/or anti permeability [interleukin (IL)-1 receptor agonist (IL-1RA), heat shock protein (HSP)-70, and adrenomedullin], proinflammatory (IL-6, IL-8, IL-2, IL-1beta, and substance P), angiogenic, or chemotactic biomarkers (macrophage inflammatory protein-1beta, VEGF, TNF-alpha, monocyte chemotactic protein-1, and matrix metalloproteinase-9) were assessed. AMS-resistant subjects had higher IL-1RA (4 and 9 h and overall), HSP-70 (0 h and overall), and adrenomedullin (overall) compared with AMS-susceptible subjects. Acetazolamide raised IL-1RA and HSP-70 compared with placebo in AMS-susceptible subjects. Dexamethasone also increased HSP-70 and adrenomedullin in AMS-susceptible subjects. Macrophage inflammatory protein-1beta was higher in AMS-susceptible than AMS-resistant subjects after 4 h of hypoxia; dexamethasone minimized this difference. Other biomarkers were unrelated to AMS. Resistance to AMS was accompanied by a marked anti-inflammatory and/or anti-permeability response that may have prevented downstream pathophysiological events leading to AMS. Conversely, AMS susceptibility does not appear to be related to an exaggerated inflammatory response. PMID- 21636567 TI - Time course of isotonic shortening and the underlying contraction mechanism in airway smooth muscle. AB - Although the structure of the contractile unit in smooth muscle is poorly understood, some of the mechanical properties of the muscle suggest that a sliding-filament mechanism, similar to that in striated muscle, is also operative in smooth muscle. To test the applicability of this mechanism to smooth muscle function, we have constructed a mathematical model based on a hypothetical structure of the smooth muscle contractile unit: a side-polar myosin filament sandwiched by actin filaments, each attached to the equivalent of a Z disk. Model prediction of isotonic shortening as a function of time was compared with data from experiments using ovine tracheal smooth muscle. After equilibration and establishment of in situ length, the muscle was stimulated with ACh (100 MUM) until force reached a plateau. The muscle was then allowed to shorten isotonically against various loads. From the experimental records, length-force and force-velocity relationships were obtained. Integration of the hyperbolic force-velocity relationship and the linear length-force relationship yielded an exponential function that approximated the time course of isotonic shortening generated by the modeled sliding-filament mechanism. However, to obtain an accurate fit, it was necessary to incorporate a viscoelastic element in series with the sliding-filament mechanism. The results suggest that a large portion of the shortening is due to filament sliding associated with muscle activation and that a small portion is due to continued deformation associated with an element that shows viscoelastic or power-law creep after a step change in force. PMID- 21636568 TI - Giant sucking sound: can physiology fill the intellectual void left by the reductionists? AB - Molecular reductionism has so far failed to deliver the broad-based therapeutic insights that were initially hoped for. This form of reductionism is now being replaced by so-called "systems biology." This is a nebulously defined approach and/or discipline, with some versions of it relying excessively on hypothesis neutral approaches and only minimally informed by key physiological concepts such as homeostasis and regulation. In this context, physiology is uniquely positioned to continue to provide impressive levels of both biological and therapeutic insight by using hypothesis-driven "classical" approaches and concepts to help frame what might be described as the "pieces of the puzzle" that emerge from molecular reductionism. The strength of physiology as a "bridge" between reductionism and epidemiology, along with its unparalleled ability to generate therapeutic insights and opportunities justifies increased attention and emphasis on our discipline into the future. Arguments relevant to this set of assertions are advanced and this paper, which was based on the 2011 Adolph Lecture, represents an effort to fill the intellectual void left by reductionism and improve scientific progress. PMID- 21636569 TI - Immunoreactive and bioactive growth hormone responses to resistance exercise in men who are lean or obese. AB - It has been suggested that obese individuals have a blunted growth hormone (GH) response to spontaneous and stimulated GH secretion. The present study was designed to examine the effects of a high-volume, whole body acute resistance exercise (RE) protocol on immunoreactive GH (iGH), bioactive GH (bGH), and GH binding protein (GHBP) in sedentary lean and obese men. Nine obese (mean +/- SD: 20.8 +/- 2.1 yr old, 177.0 +/- 4.1 cm height, 108.7 +/- 15.9 kg body mass, 37.6 +/- 5.29% body fat) and nine lean (20.1 +/- 2.1 yr old, 177.8 +/- 8.7 cm height, 71.7 +/- 5.8 kg body mass, 14.7 +/- 3.54% body fat) men completed an acute RE protocol (6 exercises, 3 sets of 10 repetitions at 85-95% of 10 repetitions maximum with 120- and 90-s rest periods), and blood samples were collected before, at the midpoint, and immediately after exercise and during recovery (+50, +70, and +110). In contrast to prior studies, which examined acute responses to cardiovascular exercise protocols, groups did not differ in iGH response to the exercise stimulus. However, bGH concentrations overall were significantly lower in the obese than the lean participants (P < 0.001). Additionally, obese individuals had significantly higher GHBP concentrations (P < 0.001). Results suggest that obese and lean sedentary men performing a high-volume, whole body acute RE protocol demonstrate similar increases in iGH. Blunted bGH and elevated GHBP concentrations are indicative of altered GH activity associated with obesity. Prior research findings of blunted iGH response may be attributable to RE protocols not equated on relative intensity or volume. These results underscore the complexity of pituitary biology and its related mechanisms and may have implications for exercise prescription in the treatment of obesity. PMID- 21636570 TI - Mechanisms of increase in cardiac output during acute weightlessness in humans. AB - Based on previous water immersion results, we tested the hypothesis that the acute 0-G-induced increase in cardiac output (CO) is primarily caused by redistribution of blood from the vasculature above the legs to the cardiopulmonary circulation. In seated subjects (n = 8), 20 s of 0 G induced by parabolic flight increased CO by 1.7 +/- 0.4 l/min (P < 0.001). This increase was diminished to 0.8 +/- 0.4 l/min (P = 0.028), when venous return from the legs was prevented by bilateral venous thigh-cuff inflation (CI) of 60 mmHg. Because the increase in stroke volume during 0 G was unaffected by CI, the lesser increase in CO during 0 G + CI was entirely caused by a lower heart rate (HR). Thus blood from vascular beds above the legs in seated subjects can alone account for some 50% of the increase in CO during acute 0 G. The remaining increase in CO is caused by a higher HR, of which the origin of blood is unresolved. In supine subjects, CO increased from 7.1 +/- 0.7 to 7.9 +/- 0.8 l/min (P = 0.037) when entering 0 G, which was solely caused by an increase in HR, because stroke volume was unaffected. In conclusion, blood originating from vascular beds above the legs can alone account for one-half of the increase in CO during acute 0 G in seated humans. A Bainbridge-like reflex could be the mechanism for the HR-induced increase in CO during 0 G in particular in supine subjects. PMID- 21636571 TI - Assembly of a filamin four-domain fragment and the influence of splicing variant 1 on the structure. AB - Filamins are scaffold proteins that bind to various proteins, including the actin cytoskeleton, integrin adhesion receptors, and adaptor proteins such as migfilin. Alternative splicing of filamin, largely constructed from 24 Ig-like domains, is thought to have a role in regulating its interactions with other proteins. The filamin A splice variant-1 (FLNa var-1) lacks 41 amino acids, including the last beta-strand of domain 19, FLNa(19), and the first beta-strand of FLNa(20) that was previously shown to mask a key binding site on FLNa(21). Here, we present a structural characterization of domains 18-21, FLNa(18-21), in the FLNa var-1 as well as its nonspliced counterpart. A model of nonspliced FLNa(18-21), obtained from small angle x-ray scattering data, shows that these four domains form an L shaped structure, with one arm composed of a pair of domains. NMR spectroscopy reveals that in the splice variant, FLNa(19) is unstructured whereas the other domains retain the same fold as in their canonical counterparts. The maximum dimensions predicted by small angle x-ray scattering data are increased upon migfilin binding in the FLNa(18-21) but not in the splice variant, suggesting that migfilin binding is able to displace the masking beta-strand and cause a rearrangement of the structure. Possible function roles for the spliced variants are discussed. PMID- 21636572 TI - Differential but competitive binding of Nogo protein and class i major histocompatibility complex (MHCI) to the PIR-B ectodomain provides an inhibition of cells. AB - Binding of class I MHC molecules (MHCI) to an inhibitory receptor, PIR-B, expressed on B cells and myeloid cells provides constitutive cellular inhibition, thus ensuring peripheral tolerance. Recent unexpected findings pointed to a novel inhibitory role of PIR-B in neurite regeneration through binding to three axonal outgrowth inhibitors of myelin, including Nogo. Thus, it becomes interesting to determine whether the actions of the inhibitory myelin proteins and MHCI could coexist independently or be mutually exclusive as to the PIR-B-mediated immune and neural cell inhibition. Here, we present data supporting the competition of Nogo- and MHCI-mediated inhibition where they coexist. Kinetic analyses of Nogo and MHCI binding to the whole or a part of the recombinant PIR-B ectodomain revealed that PIR-B binds with higher affinity to Nogo than MHCI and that the MHCI binding only occurred with the N-terminal domains of PIR-B, whereas Nogo binding occurred with either the N- or C-terminal ectodomains. Importantly, kinetic tests indicated that the binding to PIR-B of Nogo and MHCI was competitive. Both endogenous and exogenous Nogo intensified the PIR-B-mediated suppression of interleukin-6 release from lipopolysaccharide-stimulated wild type, but not PIR-B-deficient, cultured mast cells, indicating that PIR-B mediates Nogo-induced inhibition. Thus, we propose a novel mechanism by which PIR B-mediated regulation is achieved differentially but competitively via MHCI and Nogo in cells of the immune system. PMID- 21636573 TI - Actin cytoskeleton remodeling by the alternatively spliced isoform of PDLIM4/RIL protein. AB - RIL (product of PDLIM4 gene) is an actin-associated protein that has previously been shown to stimulate actin bundling by interacting with actin-cross-linking protein alpha-actinin-1 and increasing its affinity to filamentous actin. Here, we report that the alternatively spliced isoform of RIL, denoted here as RILaltCterm, functions as a dominant-negative modulator of RIL-mediated actin reorganization. RILaltCterm is regulated at the level of protein stability, and this protein isoform accumulates particularly in response to oxidative stress. We show that the alternative C-terminal segment of RILaltCterm has a disordered structure that directs the protein to rapid degradation in the core 20 S proteasomes. Such degradation is ubiquitin-independent and can be blocked by binding to NAD(P)H quinone oxidoreductase NQO1, a detoxifying enzyme induced by prolonged exposure to oxidative stress. We show that either overexpression of RILaltCterm or its stabilization by stresses counteracts the effects produced by full-length RIL on organization of actin cytoskeleton and cell motility. Taken together, the data suggest a mechanism for fine-tuning actin cytoskeleton rearrangement in response to stresses. PMID- 21636574 TI - Conditional inactivation of the CXCR4 receptor in osteoprecursors reduces postnatal bone formation due to impaired osteoblast development. AB - Cysteine (C)-X-C motif chemokine receptor 4 (CXCR4), the primary receptor for stromal cell-derived factor-1 (SDF-1), is involved in bone morphogenic protein 2 (BMP2)-induced osteogenic differentiation of mesenchymal progenitors. To target the in vivo function of CXCR4 in bone and explore the underlying mechanisms, we conditionally inactivated CXCR4 in osteoprecursors by crossing osterix (Osx)-Cre mice with floxed CXCR4 (CXCR4(fl/fl)) mice to generate knock-outs with CXCR4 deletion driven by the Osx promoter (Osx::CXCR4(fl/fl)). The Cre-mediated excision of CXCR4 occurred exclusively in bone of Osx::CXCR4(fl/fl) mice. When compared with littermate controls, Osx::CXCR4(fl/fl) mice developed smaller osteopenic skeletons as evidenced by reduced trabecular and cortical bone mass, lower bone mineral density, and a slower mineral apposition rate. In addition, Osx::CXCR4(fl/fl) mice displayed chondrocyte disorganization in the epiphyseal growth plate associated with decreased proliferation and collagen matrix syntheses. Moreover, mature osteoblast-related expression of type I collagen alpha1 and osteocalcin was reduced in bone of Osx::CXCR4(fl/fl) mice versus controls, suggesting that CXCR4 deficiency results in arrested osteoblast progression. Primary cultures for osteoblastic cells derived from Osx::CXCR4(fl/fl) mice also showed decreased proliferation and impaired osteoblast differentiation in response to BMP2 or BMP6 stimulation, and suppressed activation of intracellular BMP receptor-regulated Smads (R-Smads) and Erk1/2 was identified in CXCR4-deficient cells and bone tissues. These findings provide the first in vivo evidence that CXCR4 functions in postnatal bone development by regulating osteoblast development in cooperation with BMP signaling. Thus, CXCR4 acts as an endogenous signaling component necessary for bone formation. PMID- 21636575 TI - Isotopomer profiling of Leishmania mexicana promastigotes reveals important roles for succinate fermentation and aspartate uptake in tricarboxylic acid cycle (TCA) anaplerosis, glutamate synthesis, and growth. AB - Leishmania parasites proliferate within nutritionally complex niches in their sandfly vector and mammalian hosts. However, the extent to which these parasites utilize different carbon sources remains poorly defined. In this study, we have followed the incorporation of various (13)C-labeled carbon sources into the intracellular and secreted metabolites of Leishmania mexicana promastigotes using gas chromatography-mass spectrometry and (13)C NMR. [U-(13)C]Glucose was rapidly incorporated into intermediates in glycolysis, the pentose phosphate pathway, and the cytoplasmic carbohydrate reserve material, mannogen. Enzymes involved in the upper glycolytic pathway are sequestered within glycosomes, and the ATP and NAD(+) consumed by these reactions were primarily regenerated by the fermentation of phosphoenolpyruvate to succinate (glycosomal succinate fermentation). The initiating enzyme in this pathway, phosphoenolpyruvate carboxykinase, was exclusively localized to the glycosome. Although some of the glycosomal succinate was secreted, most of the C4 dicarboxylic acids generated during succinate fermentation were further catabolized in the TCA cycle. A high rate of TCA cycle anaplerosis was further suggested by measurement of [U-(13)C]aspartate and [U (13)C]alanine uptake and catabolism. TCA cycle anaplerosis is apparently needed to sustain glutamate production under standard culture conditions. Specifically, inhibition of mitochondrial aconitase with sodium fluoroacetate resulted in the rapid depletion of intracellular glutamate pools and growth arrest. Addition of high concentrations of exogenous glutamate alleviated this growth arrest. These findings suggest that glycosomal and mitochondrial metabolism in Leishmania promastigotes is tightly coupled and that, in contrast to the situation in some other trypanosomatid parasites, the TCA cycle has crucial anabolic functions. PMID- 21636576 TI - Impaired nutrient signaling and body weight control in a Na+ neutral amino acid cotransporter (Slc6a19)-deficient mouse. AB - Amino acid uptake in the intestine and kidney is mediated by a variety of amino acid transporters. To understand the role of epithelial neutral amino acid uptake in whole body homeostasis, we analyzed mice lacking the apical broad-spectrum neutral (0) amino acid transporter B(0)AT1 (Slc6a19). A general neutral aminoaciduria was observed similar to human Hartnup disorder which is caused by mutations in SLC6A19. Na(+)-dependent uptake of neutral amino acids into the intestine and renal brush-border membrane vesicles was abolished. No compensatory increase of peptide transport or other neutral amino acid transporters was detected. Mice lacking B(0)AT1 showed a reduced body weight. When adapted to a standard 20% protein diet, B(0)AT1-deficient mice lost body weight rapidly on diets containing 6 or 40% protein. Secretion of insulin in response to food ingestion after fasting was blunted. In the intestine, amino acid signaling to the mammalian target of rapamycin (mTOR) pathway was reduced, whereas the GCN2/ATF4 stress response pathway was activated, indicating amino acid deprivation in epithelial cells. The results demonstrate that epithelial amino acid uptake is essential for optimal growth and body weight regulation. PMID- 21636577 TI - Interaction of the HIV-1 gp120 viral protein V3 loop with bacterial lipopolysaccharide: a pattern recognition inhibition. AB - HIV-1 represents an elusive target for therapeutic compounds due to its high rate of mutation. Targeting structural patterns instead of a constantly changing specific three-dimensional structure may represent an approach that is less sensitive to viral mutations. The V3 loop of gp120 of HIV-1, which is responsible for binding of viral gp120 to CCR5 or CXCR4 coreceptors, has already been identified as an effective target for the inhibition of viral entry. The peptide derived from the V3 loop of gp120 specifically interacts with the lipid A moiety of LPS, as does the full gp120 protein. NMR analysis of V3 in complex with LPS shows formation of an amphipathic turn. The interaction between LPS and V3 relies on the structural pattern, comprising a combination of hydrophobic and charge interactions, similar to the interaction between antimicrobial peptides and LPS. LPS inhibited binding of gp120 to the surface of target T cells. Nonendotoxic LPS antagonists inhibited viral infection, demonstrating the possibility for the development of an inhibitor of HIV-1 attachment to T cells based on the recognition of a conserved structural pattern. PMID- 21636578 TI - Inhibition of RNase L and RNA-dependent protein kinase (PKR) by sunitinib impairs antiviral innate immunity. AB - RNase L and RNA-dependent protein kinase (PKR) are effectors of the interferon antiviral response that share homology in their pseudokinase and protein kinase domains, respectively. Sunitinib is an orally available, ATP-competitive inhibitor of VEGF and PDGF receptors used clinically to suppress angiogenesis and tumor growth. Sunitinib also impacts IRE1, an endoplasmic reticulum protein involved in the unfolded protein response that is closely related to RNase L. Here, we report that sunitinib is a potent inhibitor of both RNase L and PKR with IC(50) values of 1.4 and 0.3 MUM, respectively. In addition, flavonol activators of IRE1 inhibited RNase L. Sunitinib treatment of wild type (WT) mouse embryonic fibroblasts resulted in about a 12-fold increase in encephalomyocarditis virus titers. However, sunitinib had no effect on encephalomyocarditis virus growth in cells lacking both PKR and RNase L. Furthermore, oral delivery of sunitinib in WT mice resulted in 10-fold higher viral titers in heart tissues while suppressing by about 2-fold the IFN-beta levels. In contrast, sunitinib had no effect on viral titers in mice deficient in both RNase L and PKR. Also, sunitinib reduced mean survival times from 12 to 6 days in virus-infected WT mice while having no effect on survival of mice lacking both RNase L and PKR. Results indicate that sunitinib treatments prevent antiviral innate immune responses mediated by RNase L and PKR. PMID- 21636579 TI - Repeat motif-containing regions within thyroglobulin. AB - Thyroglobulin (precursor for thyroid hormone synthesis) is a large secreted glycoprotein comprising contiguous region I (multiple type-1 repeating units engaging the first ~1,191 residues, followed by a ~245-residue hinge region), regions II-III (multiple type-2 and 3 repeating units, comprising ~720 residues), and the C-terminal cholinesterase-like (ChEL) domain (~570 residues). A signal peptide attached to ChEL makes an independent secretory protein that binds to I II-III, stabilizing it and rescuing the secretion of I-II-III that would otherwise be trapped in the endoplasmic reticulum (ER). In this study, we found that a signal peptide attached to regions II-III also makes for an efficient secretory protein that neither demonstrably interacts nor has its secretion enhanced by the presence of secretory ChEL. By contrast, region I, either with or without the hinge region, cannot be secreted on its own and remains in the ER where it is bound to ER chaperones BiP and GRP94. Whereas ChEL can rescue secretion of I-II-III, it can rescue I-II only very weakly, and region I not at all. Yet, ChEL begins to rescue region I in cells that also co-express secretory II-III. The data suggest that conformational maturation of region I is a limiting step in the thyroglobulin maturation process, and this step is facilitated by the presence of both regions II-III and ChEL. Mutations causing hypothyroidism might induce solely local/regional misfolding or may interfere more globally by impeding interactions between regions that are required for thyroglobulin secretion. PMID- 21636580 TI - A unique nail gun injury to the heart with a delayed presentation. AB - We describe a 24-year-old construction worker who was unaware that he had been shot by a pneumatic nail gun in the chest during work. After returning home, he felt some palpitations and mild shortness of breath, and in the mirror discovered a non-bleeding pinpoint skin wound in his upper chest. He admitted himself to the emergency department of a local hospital and, after a detailed history and a chest X-ray had been taken, he was surprisingly diagnosed with a penetrating nail injury to the heart and was referred to our center. Transthoracic echocardiography and chest computed tomography were done, and the patient was transported to the operating room. After the nail had been removed and the mitral valve repaired, the patient was discharged on the fifth postoperative day without any complications. PMID- 21636581 TI - Reliability of repeated cognitive assessment of dementia using a brief computerized battery. AB - OBJECTIVE: The aim of this study was to evaluate the short-term stability and reliability of a brief computerized cognitive battery in established dementia types. METHOD: Patients were administered the computerized battery twice with administrations approximately 2 hours apart, with intervening conventional neuropsychological tests. Patients were classified clinically, via consensus conference, as healthy controls (n = 23), mild cognitive impairment (n = 20), Alzheimer's disease (n = 52), dementia with Lewy Bodies ([DLB], n = 10), or frontotemporal dementia (n = 9). RESULTS: Minimal practice effects were evident across Cog-State test administrations. Small magnitude improvements were seen across all groups on a working memory task, and healthy controls showed a mild practice effect on the accuracy of associative learning. CONCLUSIONS: In established dementia, administration of the CogState tasks appears sensitive to cognitive impairment in dementia. Repeat administration also provided acceptable stability and test-retest reliability with minimal practice effects at short test retest intervals despite intervening cognitive challenges. PMID- 21636582 TI - Understanding caregiver health behaviors: depressive symptoms mediate caregiver self-efficacy and health behavior patterns. AB - Previous research on female caregivers of elderly relatives with dementia has demonstrated that caregiving self-efficacy (SE) is associated with reduced cumulative health risk. The overarching aim of the current study was to expand on that research by exploring whether depressive symptoms mediate the relationship between 3 domains of caregiving SE and cumulative health risk associated with health behavior patterns. Data from 256 female family caregivers of patients with dementia are presented. Path analysis revealed a significant mediated effect for depressive symptoms as both SE for obtaining respite and SE for controlling upsetting Thoughts had a significant, indirect effect on cumulative health risk. There were no direct effects between caregiver SE and cumulative health risk. The current study sheds light on the complex pathway between caregiver SE and health and speaks to the importance of skills-based interventions designed to enhance efficacy beliefs and minimize depression in dementia caregivers. PMID- 21636584 TI - Association of IS5 with divergent tandem blaCMY-2 genes in clinical isolates of Escherichia coli. AB - OBJECTIVE: To characterize a unique tandem bla(CMY-2) gene arrangement found in two non-identical clinical strains of Escherichia coli. METHODS: Both plasmid and chromosomal DNA were evaluated using PFGE, restriction digest analysis, plasmid profiling and Southern hybridization. bla(CMY-2) gene expression and gene copy number were evaluated by real-time PCR. Susceptibilities to selected beta-lactam antibiotics were determined by agar dilution. RESULTS: A tandem arrangement for bla(CMY-2) was identified in both isolates and was the only arrangement for bla(CMY-2) observed. These isolates had distinct PFGE and plasmid profiles. Each strain exhibited 2-fold higher bla(CMY-2) mRNA expression and up to 8-fold lower beta-lactam susceptibility compared with a strain with a single copy of bla(CMY 2). CONCLUSION: This is the first report of IS5 being associated with tandem bla(CMY-2). IS5 has previously been associated with antibiotic resistance through tandem gene amplification. The unique tandem arrangement provides a mechanism for increased bla(CMY-2) expression. PMID- 21636583 TI - Comparative effectiveness of continuing a virologically effective first-line boosted protease inhibitor combination or of switching to a three-drug regimen containing either efavirenz, nevirapine or abacavir. AB - OBJECTIVES: To compare virological effectiveness in patients who continued on a virologically successful first-line boosted protease inhibitor (PI)-containing combination antiretroviral therapy (cART) regimen or who switched to a PI-free cART including efavirenz, nevirapine or abacavir. METHODS: From the French Hospital Database on HIV, we selected 439 patients with undetectable viral load (VL) on a first-line boosted PI-containing cART regimen who switched to a PI-free combination including efavirenz, nevirapine or abacavir. Each of these patients was matched with three patients who continued to take their first-line cART regimen, on the basis of gender, age, CD4 cell count, VL, date of cART initiation and the duration of VL undetectability. Time to virological failure (VF) was analysed with Kaplan-Meier curves and Cox models. RESULTS: The 12 month probabilities of VF were 3.7% and 5.7% in non-switch and switch patients, respectively, and 3.9%, 7.2% and 9.0% in patients switching to efavirenz-, nevirapine- and abacavir-containing cART, respectively. After adjustment, only patients switching to abacavir-containing cART had a higher risk of VF than non switch patients (adjusted hazard ratio, 1.99; 95% confidence interval, 1.05 3.79). CONCLUSIONS: Switching from a virologically successful first-line boosted PI-containing cART regimen to a non-nucleoside reverse transcriptase inhibitor containing cART regimen containing either efavirenz or nevirapine is virologically safe, while switching to abacavir-containing cART should be avoided. PMID- 21636585 TI - Life-threatening acute generalized exanthematous pustulosis induced by two different protease inhibitors in an HIV-1-infected patient. PMID- 21636586 TI - Molecular analysis of ciprofloxacin resistance and clonal relatedness of clinical Escherichia coli isolates from haematology patients receiving ciprofloxacin prophylaxis. AB - OBJECTIVES: Widespread use of fluoroquinolones has led to increased levels of resistance in clinical isolates of Escherichia coli. We investigated the evolution of ciprofloxacin susceptibility and molecular epidemiology of clinical E. coli isolates in haematology patients receiving ciprofloxacin prophylaxis on the population and individual patient level. METHODS: From August 2006 through December 2007 we collected all E. coli isolates (n = 404) from surveillance and infection-site cultures from 169 haematology patients receiving ciprofloxacin prophylaxis. Analysis of the gyrase A (gyrA) gene was performed by denaturing gradient gel electrophoresis (DGGE) in 364 isolates and clonal relatedness was determined by the single-enzyme amplified fragment length polymorphism (seAFLP) technique in 162 isolates. One hundred of these isolates were also subjected to qnrA analysis. RESULTS: The average number of samples per patient was 2.4 (maximum 20) and 122 (30%) of 404 E. coli isolates were resistant to ciprofloxacin. In 124 patients only ciprofloxacin-susceptible strains were detected. DGGE revealed 11 different gyrA sequence patterns and, based on AFLP analysis, there was evidence of selection of ciprofloxacin-resistant strains under antibiotic pressure, as well as the occurrence of genetically indistinguishable ciprofloxacin-resistant and -susceptible E. coli isolates within one patient. Clonal dissemination of ciprofloxacin-resistant E. coli was observed, but did not predominate. CONCLUSIONS: The genetic evolution of clinical E. coli isolates in haematology patients receiving ciprofloxacin prophylaxis is characterized by selection of ciprofloxacin-resistant strains. However, we did find evidence for de novo resistance mutation in ciprofloxacin-susceptible E. coli in individual patients under selective pressure. PMID- 21636587 TI - Hospice use among African Americans, Asians, Hispanics, and Whites: implications for practice. AB - OBJECTIVE: This study examined the characteristics of individuals in hospice care by racial/ethnic groups. METHODS: A total of 22,936 patients served by a hospice in Central Florida during a four-year period, from 2002 to 2006, were included. Of these, 80.6% were White, 9.6% were Black/African-American, 9.3% were Hispanic and 0.5% were Asian American/Pacific Islander. We examined the associations between the characteristics of hospice users and race/ethnicity, and change of hospice user characteristics over time using chi-square and ANOVA tests. RESULTS: More females than males were represented. Spouse caregivers were most common for Whites (35%) and Asian/Pacific Islanders (36%). However, "other" (41%) caregivers were most frequent for African Americans and daughters (33%) were most often caregivers for Hispanics. Cancer was the primary diagnosis across the four groups. Racial/ethnic minorities were more likely to rely on Medicaid than Whites (10-70% vs. 4%) and African Americans were most likely to be transferred from hospital (57%), whereas Whites were referred from assisted living/nursing homes more frequently than others(16% vs. 7-10%). CONCLUSION: As the hospice settings become more racially/ethnically diverse, it is essential to attend to the different circumstances and needs of the various groups in providing optimal care. PMID- 21636588 TI - Cultural competency and diversity among hospice palliative care volunteers. AB - This case study examines the current state of cultural competence in hospice and palliative care in the Greater Toronto Area (GTA). Because of changing demographic trends and ethnic minorities underutilizing hospice palliative care services, this research examined the current state of culturally competent care in a hospice setting, and the challenges to providing culturally competent care in a hospice in the GTA. A case study was conducted with a hospice and included in-depth interviews with 14 hospice volunteers. The findings reveal that volunteers encountered cultural clashes when their level of cultural competency was weak. Second, volunteers revealed there was a lack of adequate cultural competency training with their hospice, and finally, there was a lack of ethnic, cultural, and linguistic diversity among the hospice volunteers. PMID- 21636589 TI - Coverage tradeoffs and power estimation in the design of whole-genome sequencing experiments for detecting association. AB - MOTIVATION: Whole-genome sequencing (WGS) allows direct interrogation of previously undetected uncommon or rare variants, which potentially contribute to the missing heritability of human disease. However, cost of sequencing large numbers of samples limits its application in case-control association studies. Here, we describe theoretical and empirical design considerations for such sequencing studies, aimed at maximizing the power of detecting association under the constraint of study-wide cost. RESULTS: We consider two cost regimes. First, assuming cost is proportional to the total amount of base pairs to be sequenced across all samples, which is a practical model for whole-genome sequencing, we explored the tradeoff in terms of study power between increasing the number of subjects and increasing depth coverage. We demonstrate that the optimal power of detecting association is achieved at medium depth coverage under a wide range of realistic conditions for case-only sequencing designs. Second, if cost is fixed per sample, which is approximately the case in exome sequencing, we show that in a simple case+control sequencing study, the optimal design should include cases totaling 1/e of all subjects. AVAILABILITY: A web tool implementing the methods is available at http://www.cs.columbia.edu/~itsik/OPERA/. PMID- 21636590 TI - Identification of cavities on protein surface using multiple computational approaches for drug binding site prediction. AB - MOTIVATION: Protein-ligand binding sites are the active sites on protein surface that perform protein functions. Thus, the identification of those binding sites is often the first step to study protein functions and structure-based drug design. There are many computational algorithms and tools developed in recent decades, such as LIGSITE(cs/c), PASS, Q-SiteFinder, SURFNET, and so on. In our previous work, MetaPocket, we have proved that it is possible to combine the results of many methods together to improve the prediction result. RESULTS: Here, we continue our previous work by adding four more methods Fpocket, GHECOM, ConCavity and POCASA to further improve the prediction success rate. The new method MetaPocket 2.0 and the individual approaches are all tested on two datasets of 48 unbound/bound and 210 bound structures as used before. The results show that the average success rate has been raised 5% at the top 1 prediction compared with previous work. Moreover, we construct a non-redundant dataset of drug-target complexes with known structure from DrugBank, DrugPort and PDB database and apply MetaPocket 2.0 to this dataset to predict drug binding sites. As a result, >74% drug binding sites on protein target are correctly identified at the top 3 prediction, and it is 12% better than the best individual approach. AVAILABILITY: The web service of MetaPocket 2.0 and all the test datasets are freely available at http://projects.biotec.tu-dresden.de/metapocket/ and http://sysbio.zju.edu.cn/metapocket. PMID- 21636591 TI - PathVisio-MIM: PathVisio plugin for creating and editing Molecular Interaction Maps (MIMs). AB - MOTIVATION: A plugin for the Java-based PathVisio pathway editor has been developed to help users draw diagrams of bioregulatory networks according to the Molecular Interaction Map (MIM) notation. Together with the core PathVisio application, this plugin presents a simple to use and cross-platform application for the construction of complex MIM diagrams with the ability to annotate diagram elements with comments, literature references and links to external databases. This tool extends the capabilities of the PathVisio pathway editor by providing both MIM-specific glyphs and support for a MIM-specific markup language file format for exchange with other MIM-compatible tools and diagram validation. AVAILABILITY: The PathVisio-MIM plugin is freely available and works with versions of PathVisio 2.0.11 and later on Windows, Mac OS X and Linux. Information about MIM notation and the MIMML format is available at http://discover.nci.nih.gov/mim. The plugin, along with diagram examples, instructions and Java source code, may be downloaded at http://discover.nci.nih.gov/mim/mim_pathvisio.html. PMID- 21636592 TI - The FAF-Drugs2 server: a multistep engine to prepare electronic chemical compound collections. AB - SUMMARY: The FAF-Drugs2 server is a web application that prepares chemical compound libraries prior to virtual screening or that assists hit selection/lead optimization before chemical synthesis or ordering. The FAF-Drugs2 web server is an enhanced version of the FAF-Drugs2 package that now includes Pan Assay Interference Compounds detection. This online toolkit has been designed through a user-centered approach with emphasis on user-friendliness. This is a unique online tool allowing to prepare large compound libraries with in house or user defined filtering parameters. AVAILABILITY: The FAF-Drugs2 server is freely available at http://bioserv.rpbs.univ-paris-diderot.fr/FAF-Drugs/. PMID- 21636593 TI - A memory-efficient data structure representing exact-match overlap graphs with application for next-generation DNA assembly. AB - MOTIVATION: Exact-match overlap graphs have been broadly used in the context of DNA assembly and the shortest super string problem where the number of strings n ranges from thousands to billions. The length l of the strings is from 25 to 1000, depending on the DNA sequencing technologies. However, many DNA assemblers using overlap graphs suffer from the need for too much time and space in constructing the graphs. It is nearly impossible for these DNA assemblers to handle the huge amount of data produced by the next-generation sequencing technologies where the number n of strings could be several billions. If the overlap graph is explicitly stored, it would require Omega(n(2)) memory, which could be prohibitive in practice when n is greater than a hundred million. In this article, we propose a novel data structure using which the overlap graph can be compactly stored. This data structure requires only linear time to construct and and linear memory to store. RESULTS: For a given set of input strings (also called reads), we can informally define an exact-match overlap graph as follows. Each read is represented as a node in the graph and there is an edge between two nodes if the corresponding reads overlap sufficiently. A formal description follows. The maximal exact-match overlap of two strings x and y, denoted by ov(max)(x, y), is the longest string which is a suffix of x and a prefix of y. The exact-match overlap graph of n given strings of length l is an edge-weighted graph in which each vertex is associated with a string and there is an edge (x, y) of weight omega=l-|ov(max)(x, y)| if and only if omega <= lambda, where |ov(max)(x, y)| is the length of ov(max)(x, y) and lambda is a given threshold. In this article, we show that the exact-match overlap graphs can be represented by a compact data structure that can be stored using at most (2lambda 1)(2?logn?+?loglambda?)n bits with a guarantee that the basic operation of accessing an edge takes O(log lambda) time. We also propose two algorithms for constructing the data structure for the exact-match overlap graph. The first algorithm runs in O(lambdalnlogn) worse-case time and requires O(lambda) extra memory. The second one runs in O(lambdaln) time and requires O(n) extra memory. Our experimental results on a huge amount of simulated data from sequence assembly show that the data structure can be constructed efficiently in time and memory. AVAILABILITY: Our DNA sequence assembler that incorporates the data structure is freely available on the web at http://www.engr.uconn.edu/~htd06001/assembler/leap.zip PMID- 21636594 TI - LinkinPath: from sequence to interconnected pathway. AB - SUMMARY: LinkinPath is a pathway mapping and analysis tool that enables users to explore and visualize the list of gene/protein sequences through various Flash driven interactive web interfaces including KEGG pathway maps, functional composition maps (TreeMaps), molecular interaction/reaction networks and pathway to-pathway networks. Users can submit single or multiple datasets of gene/protein sequences to LinkinPath to (i) determine the co-occurrence and co-absence of genes/proteins on animated KEGG pathway maps; (ii) compare functional compositions within and among the datasets using TreeMaps; (iii) analyze the statistically enriched pathways across the datasets; (iv) build the pathway-to pathway networks for each dataset; (v) explore potential interaction/reaction paths between pathways; and (vi) identify common pathway-to-pathway networks across the datasets. AVAILABILITY: LinkinPath is freely available to all interested users at http://www.biotec.or.th/isl/linkinpath/. PMID- 21636595 TI - Aligning short reads to reference alignments and trees. AB - MOTIVATION: Likelihood-based methods for placing short read sequences from metagenomic samples into reference phylogenies have been recently introduced. At present, it is unclear how to align those reads with respect to the reference alignment that was deployed to infer the reference phylogeny. Moreover, the adaptability of such alignment methods with respect to the underlying reference alignment strategies/philosophies has not been explored. It has also not been assessed if the reference phylogeny can be deployed in conjunction with the reference alignment to improve alignment accuracy in this context. RESULTS: We assess different strategies for short read alignment and propose a novel phylogeny-aware alignment procedure. Our alignment method can improve the accuracy of subsequent phylogenetic placement of the reads into a reference phylogeny by up to 5.8 times compared with phylogeny-agnostic methods. It can be deployed to align reads to alignments generated by using fundamentally different alignment strategies (e.g. PRANK(+F) versus MUSCLE). AVAILABILITY: http://www.exelixis-lab.org/software.html PMID- 21636596 TI - Comparative studies of de novo assembly tools for next-generation sequencing technologies. AB - MOTIVATION: Several new de novo assembly tools have been developed recently to assemble short sequencing reads generated by next-generation sequencing platforms. However, the performance of these tools under various conditions has not been fully investigated, and sufficient information is not currently available for informed decisions to be made regarding the tool that would be most likely to produce the best performance under a specific set of conditions. RESULTS: We studied and compared the performance of commonly used de novo assembly tools specifically designed for next-generation sequencing data, including SSAKE, VCAKE, Euler-sr, Edena, Velvet, ABySS and SOAPdenovo. Tools were compared using several performance criteria, including N50 length, sequence coverage and assembly accuracy. Various properties of read data, including single end/paired-end, sequence GC content, depth of coverage and base calling error rates, were investigated for their effects on the performance of different assembly tools. We also compared the computation time and memory usage of these seven tools. Based on the results of our comparison, the relative performance of individual tools are summarized and tentative guidelines for optimal selection of different assembly tools, under different conditions, are provided. PMID- 21636597 TI - Robust biclustering by sparse singular value decomposition incorporating stability selection. AB - MOTIVATION: Over the past decade, several biclustering approaches have been published in the field of gene expression data analysis. Despite of huge diversity regarding the mathematical concepts of the different biclustering methods, many of them can be related to the singular value decomposition (SVD). Recently, a sparse SVD approach (SSVD) has been proposed to reveal biclusters in gene expression data. In this article, we propose to incorporate stability selection to improve this method. Stability selection is a subsampling-based variable selection that allows to control Type I error rates. The here proposed S4VD algorithm incorporates this subsampling approach to find stable biclusters, and to estimate the selection probabilities of genes and samples to belong to the biclusters. RESULTS: So far, the S4VD method is the first biclustering approach that takes the cluster stability regarding perturbations of the data into account. Application of the S4VD algorithm to a lung cancer microarray dataset revealed biclusters that correspond to coregulated genes associated with cancer subtypes. Marker genes for different lung cancer subtypes showed high selection probabilities to belong to the corresponding biclusters. Moreover, the genes associated with the biclusters belong to significantly enriched cancer-related Gene Ontology categories. In a simulation study, the S4VD algorithm outperformed the SSVD algorithm and two other SVD-related biclustering methods in recovering artificial biclusters and in being robust to noisy data. AVAILABILITY: R-Code of the S4VD algorithm as well as a documentation can be found at http://s4vd.r forge.r-project.org/. PMID- 21636598 TI - Loading and maintenance dose algorithms for phenprocoumon and acenocoumarol using patient characteristics and pharmacogenetic data. AB - AIMS: Polymorphisms in CYP2C9 and VKORC1 influence patients' phenprocoumon (PHE) and acenocoumarol (ACE) dose requirements. To provide physicians with tools to estimate the patient's individual dose, we aimed to develop algorithms for PHE and ACE. METHODS AND RESULTS: In two Dutch anticoagulation clinics, data on age, sex, height, weight, co-medication, coumarin derivative doses, and international normalized ratio values were obtained from 624 patients taking PHE and 471 taking ACE. Single nucleotide polymorphisms relevant to coumarin derivative dosing on the CYP2C9 and VKORC1 genes were determined. Using multiple linear regression, we developed genotype-guided and non-genotype-guided algorithms to predict the maintenance dose with patient characteristics and genetic information. In addition, loading doses were derived from the calculated maintenance doses. We performed external validation in an independent data set with 229 PHE and 168 ACE users. CYP2C9 and VKORC1 genotype, weight, height, sex, age, and amiodarone use contributed to the maintenance dose of PHE and ACE. The genotype-guided algorithms explained 55.9% (PHE) and 52.6% (ACE) of the variance of the maintenance dose, the non-genetic algorithms 17.3% (PHE) and 23.7% (ACE). Validation in an independent data set resulted in an explained variation of 59.4% (PHE) and 49.0% (ACE) for the genotype-guided algorithms and for 23.5% (PHE) and 17.8% (ACE) for the non-genotype-guided algorithms, without height and weight as parameters. CONCLUSION: To our knowledge, these are the first genotype-guided loading and maintenance dose algorithms for PHE and ACE using large cohorts. The utility of these algorithms will be tested in randomized controlled trials. PMID- 21636599 TI - Prevention of dementia should start 20 years before symptoms become apparent. PMID- 21636602 TI - Test anxiety in relation to measures of cognitive and intellectual functioning. AB - The potential impact of test anxiety on cognitive testing was examined in a sample of 300 predominantly male veteran referrals who were administered a comprehensive neuropsychological test battery. Exclusionary criteria included failure on effort testing (n= 14). Level of test anxiety was significantly related to performance on the WAIS-III Working Memory Index (r = -.343, p < .001) but not to scores on the Processing Speed, Perceptual Organization, or Verbal Comprehension indexes. Test anxiety was not related to a global index of neuropsychological performance on the HRNES-R (Average Impairment Scale). Level of education had a collinear relationship with test anxiety in predicting cognitive test performance. Regression analyses revealed a more prominent role for education, indicating the possibility that test anxiety may be a reaction to, more than a cause of, deficient working memory performance. These results suggest that clinicians who use these particular tests should be reluctant to attribute poor test performance to anxiety that occurs during the testing process. PMID- 21636603 TI - Identification of tammar wallaby SIRH12, derived from a marsupial-specific retrotransposition event. AB - In humans and mice, there are 11 genes derived from sushi-ichi related retrotransposons, some of which are known to play essential roles in placental development. Interestingly, this family of retrotransposons was thought to exist only in eutherian mammals, indicating their significant contributions to the eutherian evolution, but at least one, PEG10, is conserved between marsupials and eutherians. Here we report a novel sushi-ichi retrotransposon-derived gene, SIRH12, in the tammar wallaby, an Australian marsupial species of the kangaroo family. SIRH12 encodes a protein highly homologous to the sushi-ichi retrotransposon Gag protein in the tammar wallaby, while SIRH12 in the South American short-tailed grey opossum is a pseudogene degenerated by accumulation of multiple nonsense mutations. This suggests that SIRH12 retrotransposition occurred only in the marsupial lineage but acquired and retained some as yet unidentified novel function, at least in the lineage of the tammar wallaby. PMID- 21636604 TI - Interleukin-15 and other myokines in chronic alcoholics. AB - AIMS: Interleukin (IL)-15 is highly expressed in skeletal muscle, where it exerts anabolic effects, increasing protein content in muscle fibres and promoting muscle growth. Alcoholics frequently suffer myopathy. Therefore, we analyse the behaviour of IL-15 (and other myokines, such as IL-6, IL-8 and tumour necrosis factor alpha (TNF-alpha)) in alcoholics. METHODS: These myokines and also malondialdehyde (MDA)--a lipid peroxidation product--were determined by radioimmunoanalytic techniques in blood samples of 35 chronic alcoholics and 13 age- and sex-matched controls, and compared with body composition, nutritional status, liver function, amount of ethanol and routine biochemical variables. RESULTS: IL-15, IL-6, TNF-alpha, IL-8 and MDA were all higher in alcoholics than in controls; MDA and IL-6 were clearly related with liver function impairment and short-term prognosis, whereas IL-15 was higher among those who died and was related to serum bilirubin. No relation was found between IL-15 and lean mass. CONCLUSION: IL-15 levels were higher in alcoholics than in controls, especially among those who died within 18 months after admission. They are not related with muscle mass, intensity of alcoholism or nutritional status, but only with serum bilirubin. IL-6 showed inverse correlations with liver function, intensity of alcoholism, nutritional status, left arm muscle mass and short-term mortality. PMID- 21636605 TI - Two-dimensional speckle-tracking strain echocardiography in long-term heart transplant patients: a study comparing deformation parameters and ejection fraction derived from echocardiography and multislice computed tomography. AB - AIMS: Longitudinal strain determined by speckle tracking is a sensitive parameter to detect systolic left ventricular dysfunction. In this study, we assessed regional and global longitudinal strain values in long-term heart transplants and compared deformation indices with ejection fraction as determined by transthoracic echocardiography (TTE) and multislice computed tomographic coronary angiography (MSCTA). METHODS AND RESULTS: TTE and MSCTA were prospectively performed in 31 transplant patients (10.6 years post-transplantation) and in 42 control subjects. Grey-scale apical views were recorded for speckle tracking (EchoPAC 7.0, GE) of the 16 segments of the left ventricle. The presence of coronary artery disease (CAD) was assessed by MSCTA. Strain analysis was performed in 1168 segments [496 in transplant patients (42.5%), 672 in control subjects (57.7%)]. Global longitudinal peak systolic strain was significantly lower in the transplant recipients than in the healthy population (-13.9 +/- 4.2 vs. -17.4 +/- 5.8%, P< 0.01). This was still the case after exclusion of the nine transplant patients with CAD (-14.1 +/- 4.4 vs. -17.4 +/- 5.8%, P=0.03). Transplant patients exhibited significantly lower regional strain values in 9 of the 16 segments. Left ventricular ejection fraction (%) (MSCTA/Simpsons method) was 60.7 +/- 10.1%/60.2 +/- 6.7% in transplant recipients vs. 64.7 +/- 6.4%/63.0 +/- 6.2% in the healthy population, P=ns. CONCLUSION: Even though 'healthy' heart transplants without CAD exhibit normal ejection fraction, deformation indices are reduced in this population when compared with control subjects. Our findings suggests that strain analysis is more sensitive than assessment of ejection fraction for the detection of abnormalities of systolic function. PMID- 21636606 TI - Amrubicin monotherapy for patients with previously treated advanced large-cell neuroendocrine carcinoma of the lung. AB - OBJECTIVE: No standard chemotherapy has been established yet for large-cell neuroendocrine carcinoma of the lung. Amrubicin is active for both small cell and non-small cell lung cancers, but its activity for large-cell neuroendocrine carcinoma is still unknown. METHODS: From January 2002 to December 2009, 18 patients with previously treated advanced large-cell neuroendocrine carcinoma received amrubicin monotherapy. The efficacy and toxicity of the treatment were analyzed retrospectively. RESULTS: The patients comprised 17 males and one female with a median age of 62 years (range, 51-77). Fourteen and four patients had a performance status of 0-1 and 2, respectively. Thirteen (72%) patients had received one prior chemotherapy, while the others had received two or more chemotherapies. All the patients had received platinum-based chemotherapy before the amrubicin treatment. A total of 63 cycles of amrubicin chemotherapy was administered in the 18 patients, with a median number of cycles per patient of 2.5 (range, 1-10). The median dose of amrubicin in the 63 cycles was 40 (range, 30-45) mg/m(2)/day for 3 days. Grades 3-4 neutropenia, thrombocytopenia and anemia were seen in 89, 17 and 22% of the patients, respectively. Grade 3 febrile neutropenia occurred in 33% of the patients. Non-hematological toxicity was generally mild and manageable. There were five cases of partial response, six of stable disease and six of progressive disease among the 18 patients, yielding an objective response rate of 27.7%. The median progression-free and overall survivals of the patients were 3.1 and 5.1 months, respectively. CONCLUSION: Amrubicin was potentially active against previously treated large-cell neuroendocrine carcinoma. PMID- 21636607 TI - Can we accredit hospital ethics? A tentative proposal. AB - OBJECTIVES: The objective of this research was to develop ethics accreditation standards for hospitals. RESEARCH DESIGN: Our research methods included a literature review, an expert focus group, the Delphi technique and a hospital survey. The entire process was separated into two stages: (1) the development of a draft of hospital ethics accreditation standards; and (2) conducting a nationwide hospital survey of the proposed standards. RESULTS: This study produced a tentative draft of hospital ethics accreditation standards comprised of six chapters and 62 standards based on the expert focus group and Delphi technique. The six chapters are: Medical ethics policies, regulations and leadership; The establishment and operation of a medical ethics committee; The establishment and operation of research-related ethics committees; Medical ethics education; Organisational ethical climate; and Respect for patients' rights and establishment of good hospital-patient relationships. The hospital survey indicated that the concept of an organisational ethical climate was new to most hospital managers, most hospitals disliked the idea of having a separate hospital ethics accreditation system, and small hospitals were concerned about their ability to comply with all of the standards. CONCLUSIONS: Regardless of whether hospital ethics accreditation can be a stand-alone accreditation or just part of existing hospital accreditation programmes, we hope this draft can serve as a good reference for future endeavours by hospital accreditation authorities. PMID- 21636608 TI - Assessment of children's capacity to consent for research: a descriptive qualitative study of researchers' practices. AB - BACKGROUND: In Canadian jurisdictions without specific legislation pertaining to research consent, the onus is placed on researchers to determine whether a child is capable of independently consenting to participate in a research study. Little, however, is known about how child health researchers are approaching consent and capacity assessment in practice. The aim of this study was to explore and describe researchers' current practices. METHODS: The study used a qualitative descriptive design consisting of 14 face-to-face interviews with child health researchers and research assistants in Southern Ontario. Transcribed interviews were analysed for common themes. RESULTS: Procedures for assessing capacity varied considerably from the use of age cutoffs to in-depth engagement with each child. Three key issues emerged from the accounts: (1) requirements that consent be provided by a single person thwarted researchers' abilities to support family decision-making; (2) little practical distinction was made between assessing if a child was capable, versus determining if study information had been adequately explained by the researcher; and (3) participants' perceived that review boards' requirements may conflict with what they considered ethical consent practices. CONCLUSION: The results suggest that researchers' consent and capacity knowledge and skills vary considerably. Perceived discrepancies between ethical practice and ethics boards' requirements suggest the need for dialogue, education and possibly ethics board reforms. Furthermore we propose, where appropriate, a 'family decision-making' model that allows parents and their children to consent together, thereby shifting the focus from separate assent and consent procedures to approaches that appropriately engage the child and family. PMID- 21636609 TI - Ethical evaluation of the website-based promotion and advertisements for in vitro fertilisation services in Turkey. PMID- 21636610 TI - Monoamine oxidase inhibitory activity in tobacco smoke varies with tobacco type. AB - BACKGROUND: It has been suggested that inhibition of monoamine oxidase (MAO) activity by components of cigarette smoke may impact on smoking addiction, but it is unclear to what extent the known MAO inhibitors in tobacco smoke cause this inhibition. METHODS: MAO inhibitory activity was measured in a series of tobacco particulate matter preparations from different brands of cigarette and loose-leaf tobacco commonly available in New Zealand. RESULTS: When tobacco extracts were diluted to contain a physiologically relevant nicotine concentration of 0.2 MUM, all samples tested inhibited MAO-A and MAO-B by between 4% and 12% in a standard assay. Per mg of nicotine, samples from factory-made cigarettes contained significantly less MAO inhibitory activity than did samples from loose-leaf tobacco. When inhibitory activity was calculated on a per mg of tar basis, there was no significant difference between loose-leaf tobaccos and factory-made cigarettes. CONCLUSIONS: The present study shows that the ratio of nicotine to MAO inhibitory activity varies depending on the type of tobacco product used. The roll-your-own tobaccos tested delivered more tar and more MAO inhibitory activity per mg of nicotine than the factory-made cigarettes. These findings suggest that smokers of roll-your-own tobacco may experience greater difficulty in stopping smoking. PMID- 21636611 TI - Through tobacco industry eyes: civil society and the FCTC process from Philip Morris and British American Tobacco's perspectives. AB - OBJECTIVE: To analyse the models Philip Morris (PM) and British American Tobacco (BAT) used internally to understand tobacco control non-governmental organizations (NGOs) and their relationship to the global tobacco control policy making process that resulted in the Framework Convention for Tobacco Control (FCTC). METHODS: Analysis of internal tobacco industry documents in the Legacy Tobacco Document Library. RESULTS: PM contracted with Mongoven, Biscoe, and Duchin, Inc. (MBD, a consulting firm specialising in NGO surveillance) as advisors. MBD argued that because NGOs are increasingly linked to epistemic communities, NGOs could insert themselves into the global policy-making process and influence the discourse surrounding the treaty-making process. MBD advised PM to insert itself into the policy-making process, mimicking NGO behaviour. BAT's Consumer and Regulatory Affairs (CORA) department argued that global regulation emerged from the perception (by NGOs and governments) that the industry could not regulate itself, leading to BAT advocating social alignment and self-regulation to minimise the impact of the FCTC. Most efforts to block or redirect the FCTC failed. CONCLUSIONS: PM and BAT articulated a global policy-making environment in which NGOs are key, non-state stakeholders, and as a result, internationalised some of their previous national-level strategies. After both companies failed to prevent the FCTC, their strategies began to align. Multinational corporations have continued to successfully employ some of the strategies outlined in this paper at the local and national level while being formally excluded from ongoing FCTC negotiations at the global level. PMID- 21636612 TI - Waterpipe tobacco products: nicotine labelling versus nicotine delivery. AB - BACKGROUND: Waterpipe tobacco package labelling typically indicates "0.0% tar" and "0.05% or 0.5% nicotine". OBJECTIVE: To determine the extent to which nicotine labeling is related to nicotine delivery. METHODS: 110 waterpipe smokers engaged in a 45-minute waterpipe smoking session. Puff topography and plasma nicotine were measured. Three waterpipe tobacco brands were used: Nakhla (0.5% nicotine), Starbuzz (0.05% nicotine), and Al Fakher (0.05% nicotine). Data were analyzed by one-way ANOVA. RESULTS: Topography did not differ across brands. Peak plasma nicotine varied significantly across brands. Al Fakher had the highest nicotine delivery (11.4 ng/ml) followed by Nakhla (9.8 ng/ml) and Starbuzz (5.8 ng/ml). CONCLUSIONS: Nicotine labelling on waterpipe tobacco products does not reflect delivery; smoking a brand with a "0.05% nicotine" label led to greater plasma nicotine levels than smoking a brand with a "0.5% nicotine" label. Waterpipe tobacco products should be labelled in a manner that does not mislead consumers. PMID- 21636613 TI - Epidemiology of internal contamination with polonium-210 in the London incident, 2006. AB - BACKGROUND: More than 700 UK residents were tested for possible contamination with polonium-210 ((210)Po) following the alleged poisoning of Mr Alexander Litvinenko in London in November 2006. This paper describes the epidemiology of internal contamination with the radionuclide in this group. METHODS: 11 locations in London had been identified as sufficiently environmentally contaminated with (210)Po to present a health risk to people associated with them. Public health consultant teams identified individuals at risk and offered 24-h urine testing for (210)Po excretion. Prevalence of internal contamination was estimated, and a retrospective cohort analysis was completed for each location. RESULTS: Overall 139 individuals (prevalence 0.19 (95% CI 0.13 to 0.27)) showed evidence of internal contamination with (210)Po, although none with uptakes likely to cause adverse health effects. Substantial prevalence was seen among specific hotel service staff, customers, staff and other users of a hotel bar, office and hospital staff, staff of one restaurant and residents of and visitors to the family home. Increased risks of contamination were seen for a hotel bar in association with occupational, behavioural and temporal factors. Occupational and guest exposure to contaminated areas of hotels were also associated with increased contamination risk. Nurses were more likely to become contaminated than other staff involved in direct patient care. CONCLUSIONS: Uptake of trace amounts of radionuclide in this incident was frequent. Occupational, behavioural and temporal gradients in contamination risk were mostly consistent with a priori site risk assessments. Utility of the investigation methods and findings for future accidental or deliberate environmental contamination incidents are discussed. PMID- 21636614 TI - Effectiveness of a lifestyle intervention in promoting the well-being of independently living older people: results of the Well Elderly 2 Randomised Controlled Trial. AB - BACKGROUND: Older people are at risk for health decline and loss of independence. Lifestyle interventions offer potential for reducing such negative outcomes. The aim of this study was to determine the effectiveness and cost-effectiveness of a preventive lifestyle-based occupational therapy intervention, administered in a variety of community-based sites, in improving mental and physical well-being and cognitive functioning in ethnically diverse older people. METHODS: A randomised controlled trial was conducted comparing an occupational therapy intervention and a no-treatment control condition over a 6-month experimental phase. Participants included 460 men and women aged 60-95 years (mean age 74.9 +/- 7.7 years; 53% <$12000 annual income) recruited from 21 sites in the greater Los Angeles metropolitan area. RESULTS: Intervention participants, relative to untreated controls, showed more favourable change scores on indices of bodily pain, vitality, social functioning, mental health, composite mental functioning, life satisfaction and depressive symptomatology (ps<0.05). The intervention group had a significantly greater increment in quality-adjusted life years (p<0.02), which was achieved cost-effectively (US $41218/UK L24868 per unit). No intervention effect was found for cognitive functioning outcome measures. CONCLUSIONS: A lifestyle-oriented occupational therapy intervention has beneficial effects for ethnically diverse older people recruited from a wide array of community settings. Because the intervention is cost-effective and is applicable on a wide scale basis, it has the potential to help reduce health decline and promote well being in older people. Trial Registration clinicaltrials.gov identifier: NCT0078634. PMID- 21636615 TI - Estimating the resources required in the roll-out of universal access to antiretroviral treatment in Zimbabwe. AB - OBJECTIVES: To develop projections of the resources required (person-years of drug supply and healthcare worker time) for universal access to antiretroviral treatment (ART) in Zimbabwe. METHODS: A stochastic mathematical model of disease progression, diagnosis, clinical monitoring and survival in HIV infected individuals. FINDINGS: The number of patients receiving ART is determined by many factors, including the strategy of the ART programme (method of initiation, frequency of patient monitoring, ability to include patients diagnosed before ART became available), other healthcare services (referral rates from antenatal clinics, uptake of HIV testing), demographic and epidemiological conditions (past and future trends in incidence rates and population growth) as well as the medical impact of ART (average survival and the relationship with CD4 count when initiated). The variations in these factors lead to substantial differences in long-term projections; with universal access by 2010 and no further prevention interventions, between 370 000 and almost 2 million patients could be receiving treatment in 2030-a fivefold difference. Under universal access, by 2010 each doctor will initiate ART for up to two patients every day and the case-load for nurses will at least triple as more patients enter care and start treatment. CONCLUSIONS: The resources required by ART programmes are great and depend on the healthcare systems and the demographic/epidemiological context. This leads to considerable uncertainty in long-term projections and large variation in the resources required in different countries and over time. Understanding how current practices relate to future resource requirements can help optimise ART programmes and inform long-term public health planning. PMID- 21636616 TI - The impact of genital warts: loss of quality of life and cost of treatment in eight sexual health clinics in the UK. AB - OBJECTIVES: To estimate the loss of quality of life and cost of treatment associated with genital warts seen in sexual health clinics. METHODS: A cross sectional questionnaire study and case note review of individuals with genital warts, carried out in eight sexual health clinics in England and Northern Ireland. Individuals with genital warts attending the participating clinics were invited to take part in the questionnaire study. 895 participants were recruited. A separate sample of 370 participants who had attended a participating clinic with a first visit for a first or recurrent episode of genital warts between April and June 2007 was included in the case note review. Quality of life was measured using the EQ-5D questionnaire and the cost of an episode of care was derived from the case note review. RESULTS: The weighted mean EQ-5D index score was 0.87 (95% CI 0.85 to 0.89). The weighted mean disutility was 0.056 (95% CI 0.038 to 0.074). The estimated mean loss of quality-adjusted life-years associated with an episode of genital warts was 0.018 (95% CI 0.0079 to 0.031), equivalent to 6.6 days of healthy life lost per episode. The weighted mean cost per episode of care was L94 (95% CI L84 to L104), not including the cost of a sexually transmitted infection screen. CONCLUSIONS: Genital warts have a substantial impact on the health service and the individual. This information can be utilised for economic evaluation of human papillomavirus vaccination. PMID- 21636617 TI - Determining the frequency of de novo germline mutations in DNA mismatch repair genes. AB - BACKGROUND: Carriers of a germline mutation in a DNA mismatch repair (MMR) gene- that is, persons with Lynch syndrome--have substantially high risks of colorectal (CRC), endometrial, and several other cancers. The proportion of carriers who have de novo mutations (not inherited from either parent) is not known. This study reports a case series of de novo mutations in MMR genes and estimates the frequency of de novo mutation in MMR genes using the Colon Cancer Family Registry. METHODS: Screening for germline MLH1, MSH2, MSH6, and PMS2 mutations was performed for all incident CRC cases recruited from cancer registries (population based probands) displaying microsatellite instability (MSI) or loss of expression of MMR genes by immunohistochemistry (IHC) and probands with CRC in multi-case families recruited from clinics (clinic based probands), regardless of MSI or IHC status. All relatives of probands with a pathogenic mutation who donated a blood sample underwent testing for the mutation identified in the proband. RESULTS: Of 261 probands (202 clinic based, 59 population based) with MMR gene mutations for whom it was possible to determine the origin of the mutation, six (2.3%, 95% CI 0.9% to 5.0%) were confirmed as de novo, and the remaining 255 (97.7%, 95% CI 95.0% to 99.1%) were inherited. Of the de novo mutation carriers, three were clinic based probands (1.5%, 95% CI 0.3% to 4.5%) and three were population based probands (5.1%, 95% CI 1.2% to 14.5%). Two were in MLH1, three in MSH2, and one in MSH6. CONCLUSION: De novo MMR gene mutations are uncommon causes of Lynch syndrome. PMID- 21636618 TI - Mobile encapsulated adipose tissue (MEAT) of cows and humans: a distinct nonneoplastic entity. AB - Mobile encapsulated adipose tissue can be found in both the subcutis and peritoneal cavity. The cutaneous lesions are more common and better described; they are usually designated as "mobile encapsulated lipoma" or "nodular-cystic fat necrosis." The clinical name of abacus tumor describes best the small marble like nodules that often can be moved freely through the subcutaneous tissue planes. Histopathologically, the nodules are composed of a dense fibrous capsule surrounding fat that may show varying degrees of necrosis, calcification, and lipomembranous changes. The peritoneal nodules are thought to originate from detached epiploic appendices. They are described occasionally in humans but more commonly in cows. Because these bovine mobile peritoneal bodies are clinically and histopathologically identical to the nodules in human subcutaneous tissue, the authors suggest that one can better understand both processes by comparing them. Because the lesions are reactive, not neoplastic, and necrosis is not an invariable feature, the authors suggest the neutral designation of mobile encapsulated adipose tissue. PMID- 21636619 TI - A 6-year outcome of patients who cancelled carpal tunnel surgery. AB - The long-term outcomes of patients with carpal tunnel syndrome who were scheduled for release but did not proceed to surgery were compared to patients who underwent surgery, matched on preoperative symptom scores. Both groups completed the Levine-Katz questionnaire 6 years after enrolment to our multicentre carpal tunnel syndrome outcomes database. Symptom and function scores improved for the surgical (n = 24) and non-surgical (n = 36) groups (p < 0.001). Improvement in symptom scores was greater in surgical patients compared to non-surgical patients (n = 24 matched pairs; p = 0.007) but improvement in function scores between groups was not significantly different (p = 0.13). For surgical patients, function and symptom scores improved by 6 months and were unchanged at 6 years. Patients planning surgical release can expect symptomatic and functional benefits within 6 months. Overall improvement was experienced by both groups, with a superior outcome achieved with surgery. The symptoms of carpal tunnel syndrome may improve without surgery, but further studies are needed to understand the natural history of the disorder. PMID- 21636620 TI - Biomechanical comparison of three fixation techniques used for four-corner arthrodesis. AB - Clinical results following four-corner arthrodesis vary and suggest that nonunion may be related to certain fixation techniques. The purpose of our study was to examine the displacement between the lunate and capitate following a simulated four-corner arthrodesis with the hypothesis that three types of fixation (Kirschner wires, dorsal circular plate, and a locked dorsal circular plate) would allow different amounts of displacement during simulated wrist flexion and extension. Cadaver wrists with simulated four-corner arthrodeses were loaded cyclically either to implant failure or until the lunocapitate displacement exceeded 1 mm. The locked dorsal circular plate group was significantly more stable than the dorsal circular plate and K-wire groups (p = 0.018 and p = 0.006). While these locked dorsal circular plates appear to be very stable our results are limited only to the biomechanical behavior of these fixation techniques within a cadaver model. PMID- 21636621 TI - Simplifying four-strand flexor tendon repair using double-stranded suture: a comparative ex vivo study on tensile strength and bulking. AB - We have compared a simple four-strand flexor tendon repair, the single cross stitch locked repair using a double-stranded suture (dsSCL) against two other four-strand repairs: the Pennington modified Kessler with double-stranded suture (dsPMK); and the cruciate cross-stitch locked repair with single-stranded suture (Modified Sandow). Thirty fresh frozen cadaveric flexor digitorum profundus tendons were transected and repaired with one of the core repair techniques using identical suture material and reinforced with identical peripheral sutures. Bulking at the repair site and tendon-suture junctions was measured. The tendons were subjected to linear load-to-failure testing. Results showed no significant difference in ultimate tensile strength between the Modified Sandow (36.8 N) and dsSCL (32.6 N) whereas the dsPMK was significantly weaker (26.8 N). There were no significant differences in 2 mm gap force, stiffness or bulk between the three repairs. We concluded that the simpler dsSCL repair is comparable to the modified Sandow repair in tensile strength, stiffness and bulking. PMID- 21636622 TI - Masson's disease in hand surgery: a clinicopathologic study of four cases. AB - Intravascular papillary endothelial hyperplasia (IPEH), also known as Masson's tumour, is an uncommon benign vascular lesion characterized by small multiple, endothelial-lined, papillary structures with hyaline stalks. It appears to be a reactive condition involving excessive proliferation of endothelial cells in normal blood vessels or in vascular malformations, perhaps in response to blood vessel injury or thrombosis. The lesions are small, superficial, reddish-blue nodules, usually in the head, neck or hand. Distinction from pyogenic granuloma and angiosarcoma usually requires pathological examination. Complete surgical excision is the treatment of choice. Four cases affecting the finger are presented together with histological features. PMID- 21636623 TI - The ezetimibe controversy: implications for clinical practice. AB - Low-density lipoprotein cholesterol (LDL-C) remains the primary target of lipid lowering therapy. Achieving LDL-C goals as outlined by the National Cholesterol Education Program Adult Treatment Panel III can be difficult with statins alone; therefore, adjunctive therapy is often indicated to reduce cardiovascular risk. Ezetimibe, a potent inhibitor of intestinal cholesterol absorption, has been shown to be safe, tolerable and effective at lowering LDL-C, non-high-density lipoprotein cholesterol and apolipoprotein B, each of which has been correlated with improved clinical outcomes, alone or in combination with a statin. However, because of randomized trials that demonstrated mixed results about atherosclerotic plaque regression via carotid intima-media thickness and a concern about cancer risk, ezetimibe's role in lipid therapy has been questioned. Currently, a large randomized controlled trial is in progress to answer if ezetimibe improves clinical outcomes in patients with high-risk acute coronary syndrome. A smaller trial in patients with chronic kidney disease demonstrated reduced clinical events, including myocardial infarction, stroke and revascularization for patients taking the combination of ezetimibe and simvastatin versus those taking statin or placebo alone. In this paper, we review the trials that have led to the ezetimibe controversy and then discuss the possible role of ezetimibe in specific patient populations until the results of ongoing clinical trials are known. PMID- 21636624 TI - Integrated headache care. AB - Patients with chronic or difficult to treat headaches are generally under the care of general practictioners or neurologists in private practice. Some are referred to a headache specialist for evaluation and advice. Treatment is often provided by the referring physician. An alternative is a multidisciplinary headache centre, where care is provided by different disciplines (neurology, behavioural psychology, psychiatry, psychosomatic medicine, physical therapy, sport therapy) across sectors of the healthcare system involving out- and inpatient care and treatment. This is called integrated headache care. This review summarizes experiences in integrated headache care settings in Europe and the USA, describes these settings, and reports outcome data. PMID- 21636625 TI - Association of GPX1 polymorphism, GPX activity and prostate cancer risk. AB - Prostate cancer is the second most common cancer in men worldwide. Although the aetiology of this disease remains largely unclear, several lines of evidence suggest that oxidative stress plays a role in prostate carcinogenesis. The antioxidant enzyme glutathione peroxidase 1 (GPX1) is part of the enzymatic antioxidant defence, preventing oxidative damage to DNA, proteins and lipids by detoxifying hydrogen and lipid peroxides that may contribute to prostate cancer development. Some studies indicate an association between GPX1 Pro198Leu polymorphism and an increased risk of cancer. The purpose of the present study was to determine the possible association of GPX1 Pro198Leu polymorphism and erythrocyte GPX activity with the risk of developing prostate cancer and to clarify whether erythrocyte GPX activity levels were correlated with the GPX1 Pro198Leu genotype in the Turkish population. The GPX1 Pro198Leu genotype was determined in 33 prostate cancer patients and 91 control individuals. As evident from our results, there was no difference between genotype and/or allele frequencies in prostate cancer patients and controls. No significant difference was found in GPX1 genotype or allele frequency between aggressive and non aggressive prostate cancer patients. It can be suggested with these findings that individual susceptibility of prostate cancer may be modulated by GPX1 polymorphism, but it needs further studies. PMID- 21636626 TI - Chronic polyarthritis as the first manifestation of juvenile systemic lupus erythematosus patients. AB - OBJECTIVE: To evaluate the prevalence of chronic polyarthritis in juvenile systemic lupus erythematosus (JSLE) and to describe the manifestations, treatments, and outcomes in these patients. METHODS: From January 1983 to July 2010, 5419 patients were followed up at the Pediatric Rheumatology Unit of the University Hospital and 271 (5%) of them had JSLE (American College of Rheumatology [ACR] criteria). 'Rhupus' was classified as the overlap of juvenile idiopathic arthritis (International League of Associations for Rheumatology [ILAR] criteria) and JSLE. We evaluated demographic data, polyarthritis and other clinical manifestations, disease activity and damage, laboratory exams, radiographic findings, treatments, and outcomes. RESULTS: The prevalence of chronic polyarthritis in this JSLE population was 2.6% (7/271). This articular involvement was the initial manifestation in all seven JSLE patients. The median duration of chronic polyarthritis was 11 months (range 2-15 months). Interestingly, rhupus with chronic polyarthritis and limitation of movement, presence of rheumatoid factor, autoantibodies, and/or radiographic abnormalities (juxtaarticular osteopenia, joint-space narrowing, or erosions) was evidenced in three patients. No patient had deformities of hands and feet associated with Jaccoud's arthropathy or osteonecrosis. All patients were treated with nonsteroidal anti-inflammatory drugs (NSAIDs, naproxen 10-15 mg/kg/day) when polyarthritis diagnosis was established. Prednisone and antimalarials were administered at JSLE diagnosis. The three non-responsive rhupus patients were treated in conjunction with immunosuppressive drugs (methotrexate, azathioprine, and/or cyclosporine). CONCLUSIONS: Chronic polyarthritis was a rare lupus manifestation in active pediatric patients. The interesting overlap between chronic arthritis and lupus, called rhupus suggests a new entity with a different clinical profile and a poor response to treatment with NSAIDs alone. In addition, the occurrence of this association in JSLE patients could be classified as a clinical sub-group of JSLE with possible specific genetic determinants. PMID- 21636627 TI - Association of heart structure and function abnormalities with laboratory findings in patients with systemic lupus erythematosus. AB - Conventional risk factors of coronary artery disease fail to explain the increased frequency of cardiovascular morbidity in patients with systemic lupus erythematosus (SLE). The study was conducted to determine possible association between the heart structure and function abnormalities with established prognostic value assessed by non-invasive imaging techniques and markers of autoimmune and inflammatory phenomena typical for SLE. Echocardiography and single photon emission computerized tomography (SPECT; Tc-99m-MIBI) at rest were performed in 60 SLE patients in a stable clinical condition of their disease. Laboratory evaluation included serum levels of C-reactive protein (CRP), complement C3c and C4 components and antiphospholipid antibodies (aPL). The latter included serum anticardiolipin (aCL) and anti-beta2-glycoprotein I (antibeta2GPI) antibodies, both of IgG and IgM class, and lupus anticoagulant (LA) in plasma. Echocardiography revealed pathologic thickening of valvular leaflets and/or pericardium in more than 60% of patients. Right ventricular systolic pressure (RVSP) was elevated (>30 mmHg) in 16.7%. Myocardial perfusion defects were present in 36.7% of patients, despite normal ECG recordings and a lack of clinical symptoms of myocardial ischaemia. There was a significant association between thickening of valvular leaflets and/or pericardium and high CRP and low C3c and C4 concentrations. On the other hand, increased RVSP and the presence of myocardial perfusion defects were associated with the presence of anticardiolipin and antibeta2GPI antibodies of the IgG class. Increased anticardiolipin IgG levels predicted perfusion defects in SPECT study with 100% sensitivity and 68% specificity, whereas elevated antibeta2GPI IgG levels predicted RVSP elevation (>30 mmHg) with 100% sensitivity and 78% specificity. In stable SLE patients pericardial and valve abnormalities may be associated with markers of an ongoing inflammation. Also, pulmonary systolic pressure elevation and myocardial perfusion defects are combined with elevated levels of anticardiolipin and antibeta2GPI antibodies of the IgG class. These results indicate that even clinically silent pulmonary hypertension and myocardial perfusion defects in SLE patients could be causally related to the presence of antiphospholipid antibodies. PMID- 21636628 TI - Lack of association of interleukin-18 gene promoter -607 A/C polymorphism with susceptibility to autoimmune diseases: a meta-analysis. AB - OBJECTIVE: Published data on the association between interleukin (IL)-18 gene promoter -607 A/C polymorphism and autoimmune diseases risk are inconclusive. To derive a more precise estimation of the relationship, a meta-analysis was performed. METHODS: A total of 17 studies, including six studies on type 1 diabetes (T1D), four on rheumatoid arthritis (RA), five on systemic lupus erythematosus (SLE), three on Crohn's Disease (CD) and three on ulcerative colitis (UC), were available for the meta-analysis. Meta-analysis was performed for genotypes A/A (recessive effect), genotypes A/A + A/C (dominant effect), and A allele in fixed or random-effects models. RESULTS: Overall, no significantly elevated autoimmune diseases risk was found in all genetic models when all studies were pooled into the meta-analysis. The overall odds ratios (ORs) and 95% confidence intervals (CIs) for A-allele were T1D (OR = 0.938, 95% CI = 0.757 1.162), RA (OR = 0.759, 95% CI = 0.540-1.067), SLE (OR = 0.858, 95% CI = 0.609 1.208), CD (OR = 1.159, 95% CI = 0.975-1.379) and UC (OR = 1.170, 95% CI = 0.977 1.402), respectively. In the subgroup analysis by ethnicity, there was still no significant association detected in all genetic models. CONCLUSIONS: To date, there is still not enough evidence to indicate the association of IL-18 gene promoter -607 A/C polymorphism and the development of autoimmune diseases. PMID- 21636629 TI - Re: "Job strain and the risk of depression: is reporting biased?". PMID- 21636634 TI - Are policy makers really listening? PMID- 21636633 TI - Effect of evidence based risk information on "informed choice" in colorectal cancer screening: randomised controlled trial. AB - OBJECTIVE: To compare the effect of evidence based information on risk with that of standard information on informed choice in screening for colorectal cancer. DESIGN: Randomised controlled trial with 6 months' follow-up. SETTING: German statutory health insurance scheme. PARTICIPANTS: 1577 insured people who were members of the target group for colorectal cancer screening (age 50-75, no history of colorectal cancer). INTERVENTIONS: Brochure with evidence based risk information on colorectal cancer screening and two optional interactive internet modules on risk and diagnostic tests; official information leaflet of the German colorectal cancer screening programme (control). MAIN OUTCOME MEASURE: The primary end point was "informed choice," comprising "knowledge," "attitude," and "combination of actual and planned uptake." Secondary outcomes were "knowledge" and "combination of actual and planned uptake." Knowledge and attitude were assessed after 6 weeks and combination of actual and planned uptake of screening after 6 months. RESULTS: The response rate for return of both questionnaires was 92.4% (n = 1457). 345/785 (44.0%) participants in the intervention group made an informed choice, compared with 101/792 (12.8%) in the control group (difference 31.2%, 99% confidence interval 25.7% to 36.7%; P < 0.001). More intervention group participants had "good knowledge" (59.6% (n = 468) v 16.2% (128); difference 43.5%, 37.8% to 49.1%; P < 0.001). A "positive attitude" towards colorectal screening prevailed in both groups but was significantly lower in the intervention group (93.4% (733) v 96.5% (764); difference -3.1%, -5.9% to -0.3%; P<0.01). The intervention had no effect on the combination of actual and planned uptake (72.4% (568) v 72.9% (577); P = 0.87). CONCLUSIONS: Evidence based risk information on colorectal cancer screening increased informed choices and improved knowledge, with little change in attitudes. The intervention did not affect the combination of actual and planned uptake of screening. Trial registration Current Controlled Trials ISRCTN47105521. PMID- 21636635 TI - US abortion rates rise among poor women but fall overall. PMID- 21636636 TI - Cause of outbreak of E coli in Germany is still uncertain. PMID- 21636637 TI - GMC suspends surgeon for plagiarism. PMID- 21636638 TI - Medical treatment, PCI, or CABG for coronary artery disease? PMID- 21636639 TI - Accelerated maturation and abnormal morphology in the preterm neonatal kidney. AB - Nephrogenesis is ongoing at the time of birth for the majority of preterm infants, but whether postnatal renal development follows a similar trajectory to normal in utero growth is unknown. Here, we examined tissue collected at autopsy from 28 kidneys from preterm neonates, whose postnatal survival ranged from 2 to 68 days, including 6 that had restricted intrauterine growth. In addition, we examined kidneys from 32 still-born gestational controls. We assessed the width of the nephrogenic zone, number of glomerular generations, cross-sectional area of the renal corpuscle, and glomerular maturity and morphology. Renal maturation accelerated after preterm birth, with an increased number of glomerular generations and a decreased width of the nephrogenic zone in the kidneys of preterm neonates. Of particular concern, compared with gestational controls, preterm kidneys had a greater percentage of morphologically abnormal glomeruli and a significantly larger cross-sectional area of the renal corpuscle, suggestive of renal hyperfiltration. These observations suggest that the preterm kidney may have fewer functional nephrons, thereby increasing vulnerability to impaired renal function in both the early postnatal period and later in life. PMID- 21636640 TI - Inhibition of the P2X7 receptor reduces cystogenesis in PKD. AB - The P2X7 receptor participates in purinergic signaling, which may promote the progression of ADPKD. We examined the effects of a P2X7 receptor antagonist and a P2X7 receptor agonist on cyst development in a zebrafish model of polycystic kidney disease in which we knocked down pkd2 by morpholinos. We used live wt-1b pronephric-specific GFP-expressing zebrafish embryos to directly observe changes in the pronephros. Exposure of pkd2-morphant zebrafish to a P2X7 receptor antagonist (oxidized ATP [OxATP]) significantly reduced the frequency of the cystic phenotype compared with either exposure to a P2X7 receptor agonist (BzATP) or with no treatment (P < 0.01). Histology confirmed improvement of glomerular cysts in OxATP-treated pkd2 morphants. OxATP also reduced p-ERK activity and cell proliferation in pronephric kidneys in pkd2 morphants. Inhibition of P2X7 with an additional specific antagonist (A-438079), and through morpholino-mediated knockdown of p2rx7, confirmed these effects. In conclusion, blockade of the P2X7 receptor reduces cyst formation via ERK-dependent pathways in a zebrafish model of polycystic kidney disease, suggesting that P2X7 antagonists may have therapeutic potential in ADPKD. PMID- 21636641 TI - Generation of induced pluripotent stem cells from urine. AB - Forced expression of selected transcription factors can transform somatic cells into embryonic stem cell (ESC)-like cells, termed induced pluripotent stem cells (iPSCs). There is no consensus regarding the preferred tissue from which to harvest donor cells for reprogramming into iPSCs, and some donor cell types may be more prone than others to accumulation of epigenetic imprints and somatic cell mutations. Here, we present a simple, reproducible, noninvasive method for generating human iPSCs from renal tubular cells present in urine. This procedure eliminates many problems associated with other protocols, and the resulting iPSCs display an excellent ability to differentiate. These data suggest that urine may be a preferred source for generating iPSCs. PMID- 21636642 TI - Hyperplastic polyposis syndrome and the risk of colorectal cancer. PMID- 21636643 TI - An 'illuminating' CT. PMID- 21636644 TI - Suboptimal gastroprotective coverage of NSAID use and the risk of upper gastrointestinal bleeding and ulcers: an observational study using three European databases. AB - BACKGROUND: Gastro-protective agents (GPA) are co-prescribed with non-steroidal anti-inflammatory drugs (NSAID) to lower the risk of upper gastrointestinal (UGI) events. It is unknown to what extent the protective effect is influenced by therapy adherence. AIM: To study the association between GPA adherence and UGI events among non-selective (ns) NSAID users. METHODS: The General Practice Research Database (UK 1998-2008), the Integrated Primary Care Information database (the Netherlands 1996-2007) and the Health Search/CSD Longitudinal Patient Database (Italy 2000-2007) were used. A nested case-control design was employed within a cohort of nsNSAID users aged >=50 years, who also used a GPA. UGI event cases (UGI bleeding and/or symptomatic ulcer with/without obstruction/perforation) were matched to event-free members of the cohort for age, sex, database and calendar time. Adherence to GPA was calculated as the proportion of nsNSAID treatment days covered by a GPA prescription. Adjusted OR with 95% CI were calculated. RESULTS: The cohort consisted of 618 684 NSAID users, generating 1 107 266 nsNSAID episodes. Of these, 117 307 (10.6%) were (partly) covered by GPA, 4.9% of which with a GPA coverage <20% (non-adherence), and 68.1% with a GPA coverage >80% (full adherence). 339 patients experienced an event. Among non-adherers, the OR was 2.39 (95% CI 1.66 to 3.44) for all UGI events and 1.89 (95% CI 1.09 to 3.28) for UGI bleeding alone, compared to full adherers. CONCLUSIONS: The risk of UGI events was significantly higher in nsNSAID users with GPA non-adherence. This underlines the importance of strategies to improve GPA adherence. PMID- 21636645 TI - The MUC13 cell-surface mucin protects against intestinal inflammation by inhibiting epithelial cell apoptosis. AB - BACKGROUND AND AIMS: The MUC13 transmembrane mucin is highly and constitutively expressed in the small and large intestine. Although MUC13 polymorphisms have been associated with human inflammatory bowel diseases and susceptibility to Escherichia coli infection in pigs, the biological functions of MUC13 are unknown. This study aimed to explore whether MUC13 modulates intestinal inflammation. METHODS: Muc13(-/-) mice were generated, phenotyped and challenged with the colitis-inducing agent, dextran sodium sulphate (DSS). Colitis was assessed by clinical symptoms and intestinal histopathology. Intestinal epithelial cell apoptosis and proliferation, macrophage infiltration and cytokine production were also quantified. Apoptosis of human LS513 intestinal epithelial cells in response to apoptotic agents, including DSS, was also measured, following knockdown of MUC13 with siRNA. RESULTS: Muc13(-/-) mice were viable, fertile and developed normally, with no spontaneous intestinal pathology except mild focal neutrophilic inflammation in the small and large intestines of old mice. In response to DSS challenge, Muc13(-/-) mice developed more severe acute colitis, as reflected by increased weight loss, rectal bleeding, diarrhoea and histological colitis scores compared with wild-type mice. Increased numbers of F4/80(+) macrophages in inflamed mucosa of Muc13(-/-) mice were accompanied by increased expression of intestinal IL-1beta and TNFalpha mRNA. Muc13(-/-) mice had significantly increased intestinal epithelial cell apoptosis within 3 days of DSS exposure. LS513 cells were more susceptible to DSS, actinomycin-D, ultraviolet irradiation and TRAIL-induced apoptosis when MUC13 was knocked down by siRNA. CONCLUSIONS: These novel findings indicate a protective role for Muc13 in the colonic epithelium by inhibiting toxin-induced apoptosis and have important implications for intestinal infections, inflammatory diseases and the development of intestinal cancer. PMID- 21636646 TI - Association of TNFSF15 polymorphism with irritable bowel syndrome. AB - BACKGROUND: Irritable bowel syndrome (IBS) is the most common gastrointestinal disorder, affecting more than 10% of the general population worldwide. Although a genetic component is suspected, unambiguous susceptibility genes have so far not been identified. This study tested the hypothesis that genes contributing to epithelial barrier integrity, control of mucosal immune responses and interactions with bacteria in the gut are associated with IBS. METHODS: Single nucleotide polymorphisms (SNPs) corresponding to top signals of association with Crohn's disease at 30 known susceptibility loci were tested for their effect on IBS risk in 1992 individuals from two independent case-control cohorts from Sweden and the USA. Association tests included a conservative Bonferroni correction for multiple comparisons, and were also performed on specific subgroups of patients characterised by constipation (IBS-C), diarrhoea (IBS-D) or alternating constipation and diarrhoea (IBS-A). RESULTS: The Crohn's disease risk allele rs4263839 G in the TNFSF15 gene was significantly associated with an increased risk of both IBS (p=2.2*10(-5); OR 1.37) and more pronouncedly, IBS-C (p=8.7*10(-7); OR 1.79) in the entire sample. Similar associations and risk effects of the same magnitude were observed in the two cohorts analysed separately. A correlation between rs4263839 genotype and TNFSF15 mRNA expression was detected both in peripheral blood and in rectal mucosal biopsies from healthy individuals (combined p=0.0033). CONCLUSIONS: TNFSF15 is a susceptibility gene for IBS and IBS constipation. As TL1A, the protein encoded by TNFSF15, contributes to the modulation of inflammatory responses, the results support a role of immune activation in IBS. PMID- 21636647 TI - p38 MAP kinase is a therapeutic target for hepatic encephalopathy in rats with portacaval shunts. AB - OBJECTIVE: Inflammation plays a role in neurological alterations in patients with hepatic encephalopathy (HE). Animal models of HE show neuroinflammation. Treatment with ibuprofen, a non-steroidal anti-inflammatory drug (NSAID), reduces neuroinflammation and restores cognitive and motor function in rats with HE due to portacaval shunts (PCS). This suggests that reducing neuroinflammation would improve neurological status in patients with minimal or clinical HE. NSAID induce kidney damage in patients with cirrhosis and PCS rats and are not suitable for clinical use. It is therefore necessary to look for procedures to eliminate neuroinflammation without inducing secondary effects in the kidney. Inhibition of p38 MAPK is being tested as a therapeutic target in inflammatory diseases and reduces microglial activation. This study aimed to assess whether inhibiting p38 with SB239063 reduces neuroinflammation and improves cognitive and motor function in PCS rats without affecting the kidney. RESULTS: p38 activity is increased in the brains of PCS rats and treatment with SB239063 reduces microglial activation, as well as inflammatory markers in brain (prostaglandin E2, cyclooxygenase activity, iNOS, IL-1beta, TNFalpha) and blood (prostaglandin E2 and TNFalpha). PCS rats showed increased ammonia and glutamine in the brain, which was not affected by SB239063. PCS rats showed reduced ability to learn a Y-maze conditional discrimination task, reduced motor activity and impaired motor coordination, as assessed in the rotarod. Treatment with SB239063 completely restored learning ability, motor activity and coordination in PCS rats. SB239063 did not affect creatinine or sodium levels in serum, indicating that it does not induce kidney damage. CONCLUSION: These findings suggest that reducing neuroinflammation by using inhibitors of p38 would improve the neurological status in HE without inducing secondary effects in the kidney. PMID- 21636648 TI - A conserved residue, PomB-F22, in the transmembrane segment of the flagellar stator complex, has a critical role in conducting ions and generating torque. AB - Bacterial flagellar motors exploit the electrochemical potential gradient of a coupling ion (H(+) or Na(+)) as their energy source, and are composed of stator and rotor proteins. Sodium-driven and proton-driven motors have the stator proteins PomA and PomB or MotA and MotB, respectively, which interact with each other in their transmembrane (TM) regions to form an ion channel. The single TM region of PomB or MotB, which forms the ion-conduction pathway together with TM3 and TM4 of PomA or MotA, respectively, has a highly conserved aspartate residue that is the ion binding site and is essential for rotation. To investigate the ion conductivity and selectivity of the Na(+)-driven PomA/PomB stator complex, we replaced conserved residues predicted to be near the conserved aspartate with H(+)-type residues, PomA-N194Y, PomB-F22Y and/or PomB-S27T. Motility analysis revealed that the ion specificity was not changed by either of the PomB mutations. PomB-F22Y required a higher concentration of Na(+) to exhibit swimming, but this effect was suppressed by additional mutations, PomA-N194Y or PomB-S27T. Moreover, the motility of the PomB-F22Y mutant was resistant to phenamil, a specific inhibitor for the Na(+) channel. When PomB-F22 was changed to other amino acids and the effects on swimming ability were investigated, replacement with a hydrophilic residue decreased the maximum swimming speed and conferred strong resistance to phenamil. From these results, we speculate that the Na(+) flux is reduced by the PomB-F22Y mutation, and that PomB-F22 is important for the effective release of Na(+) from PomB-D24. PMID- 21636649 TI - Diversity and distribution of transcription factors: their partner domains play an important role in regulatory plasticity in bacteria. AB - The ability of bacteria to deal with diverse environmental changes depends on their repertoire of genes and their ability to regulate their expression. In this process, DNA-binding transcription factors (TFs) have a fundamental role because they affect gene expression positively and/or negatively depending on operator context and ligand-binding status. Here, we show an exhaustive analysis of winged helix-turn-helix domains (wHTHs), a class of DNA-binding TFs. These proteins were identified in high proportions and widely distributed in bacteria, representing around half of the total TFs identified so far. In addition, we evaluated the repertoire of wHTHs in terms of their partner domains (PaDos), identifying a similar trend, as with TFs, i.e. they are abundant and widely distributed in bacteria. Based on the PaDos, we defined three main groups of families: (i) monolithic, those families with little PaDo diversity, such as LysR; (ii) promiscuous, those families with a high PaDo diversity; and (iii) monodomain, with families of small sizes, such as MarR. These findings suggest that PaDos have a very important role in the diversification of regulatory responses in bacteria, probably contributing to their regulatory complexity. Thus, the TFs discriminate over longer regions on the DNA through their diverse DNA-binding domains. On the other hand, the PaDos would allow a great flexibility for transcriptional regulation due to their ability to sense diverse stimuli through a variety of ligand-binding compounds. PMID- 21636650 TI - Biofilm formation by zygomycetes: quantification, structure and matrix composition. AB - Most studies on fungal biofilms have focused on Candida in yeasts and Aspergillus in mycelial fungi. To the authors' knowledge, biofilm formation by zygomycetes has not been reported previously. In this study, the biofilm-forming capacity of Rhizopus oryzae, Lichtheimia corymbifera, Rhizomucor pusillus and Apophysomyces elegans was evaluated. At appropriate seeding spore densities, Rhp. oryzae (105 c.f.u. ml-1, L. corymbifera (104 c.f.u. ml-1) and Rhm. pusillus (104 c.f.u. ml-1) produced highly intertwined, adherent structures on flat-bottomed polystyrene microtitre plates after 24 h at 37 degrees C. The adhered fungal hyphae were encased in an extracellular matrix, as confirmed by phase-contrast and confocal microscopy. The thickness of Rhp. oryzae, L. corymbifera and Rhm. pusillus biofilms was 109.67+/-10.02, 242+/-23.07 and 197+/-9.0 um (mean+/-sd), respectively. Biochemical characterization of the biofilm matrix indicated the presence of glucosamine, constituting 74.54-82.22 % of its dry weight, N acetylglucosamine, glucose and proteins. Adherence and biofilm formation were not observed in A. elegans. Although A. elegans spores germinated at all three seeding densities tested (1*107, 1*106 and 1*105 c.f.u. ml-1), no significant difference was observed (P>0.05) between the A490 of wells inoculated with A. elegans and the cut-off A490 for biofilm detection. This study highlights the potential for biofilm formation by at least three medically important species of zygomycetes. PMID- 21636651 TI - Cell envelope gene expression in phosphate-limited Bacillus subtilis cells. AB - The high phosphate content of Bacillus subtilis cell walls dictates that cell wall metabolism is an important feature of the PhoPR-mediated phosphate limitation response. Here we report the expression profiles of cell-envelope associated and PhoPR regulon genes, determined by live cell array and transcriptome analysis, in exponentially growing and phosphate-limited B. subtilis cells. Control by the WalRK two-component system confers a unique expression profile and high level of promoter activity on the genes of its regulon with yocH and cwlO expression differing both qualitatively and quantitatively from all other autolysin-encoding genes examined. The activity of the PhoPR two-component system is restricted to the phosphate-limited state, being rapidly induced in response to the cognate stimulus, and can be sustained for an extended phosphate limitation period. Constituent promoters of the PhoPR regulon show heterogeneous induction profiles and very high promoter activities. Phosphate-limited cells also show elevated expression of the actin-like protein MreBH and reduced expression of the WapA cell wall protein and WprA cell wall protease indicating that cell wall metabolism in this state is distinct from that of exponentially growing and stationary-phase cells. The PhoPR response is very rapidly deactivated upon removal of the phosphate limitation stimulus with concomitant increased expression of cell wall metabolic genes. Moreover expression of genes encoding enzymes involved in sulphur metabolism is significantly altered in the phosphate-limited state with distinct perturbations being observed in wild-type 168 and AH024 (DeltaphoPR) cells. PMID- 21636652 TI - Acyl depsipeptide (ADEP) resistance in Streptomyces. AB - ADEP, a molecule of the acyl depsipeptide family, has an antibiotic activity with a unique mode of action. ADEP binding to the ubiquitous protease ClpP alters the structure of the enzyme. Access of protein to the ClpP proteolytic chamber is therefore facilitated and its cohort regulatory ATPases (ClpA, ClpC, ClpX) are not required. The consequent uncontrolled protein degradation in the cell appears to kill the ADEP-treated bacteria. ADEP is produced by Streptomyces hawaiiensis. Most sequenced genomes of Streptomyces have five clpP genes, organized as two distinct bicistronic operons, clpP1clpP2 and clpP3clpP4, and a single clpP5 gene. We investigated whether the different Clp proteases are all sensitive to ADEP. We report that ClpP1 is a target of ADEP whereas ClpP3 is largely insensitive. In wild-type Streptomyces lividans, clpP3clpP4 expression is constitutively repressed and the reason for the maintenance of this operon in Streptomyces has been elusive. ClpP activity is indispensable for survival of actinomycetes; we therefore tested whether the clpP3clpP4 operon, encoding an ADEP-insensitive Clp protease, contributes to a mechanism of ADEP resistance by target substitution. We report that in S. lividans, inactivation of ClpP1ClpP2 production or protease activity is indeed a mode of resistance to ADEP although it is neither the only nor the most frequent mode of resistance. The ABC transporter SclAB (orthologous to the Streptomyces coelicolor multidrug resistance pump SCO4959-SCO4960) is also able to confer ADEP resistance, and analysis of strains with sclAB deletions indicates that there are also other mechanisms of ADEP resistance. PMID- 21636653 TI - The use of the PleurX catheter in the management of non-malignant pleural effusions. AB - PURPOSE: To evaluate the effectiveness of the PleurX catheter in the management of recurrent non-malignant pleural effusions. METHODS: All subjects who underwent a PleurX catheter placement between 2003 and 2009 were evaluated. General demographic data, time to pleurodesis, complications, and a satisfaction questionnaire were collected. The subjects were divided into two groups. Group I included patients with non-malignant effusions and group II included patients with malignant effusions. RESULTS: A total of 64 subjects were included in the final data analysis. A total of 23 subjects were included in group I and 41 subjects were included in group II. The diagnoses in group I included congestive heart failure (CHF; 13), hepatic hydrothorax (8), traumatic bloody (1), and idiopathic exudative (1). The diagnoses in group II included lung cancer (20), breast cancer (11), colon cancer (5), prostate cancer (2), B-cell lymphoma (2), and mesothelioma (1). The time to pleurodesis was 36 +/- 12 days for group II compared to 110.8 +/- 41 days for group I (p < 0.0001). The mean satisfaction score was similar in both groups (3.8 +/- 0.4). Time to pleurodesis was significantly shorter in hepatic hydrothorax compared to CHF (73.6 +/- 9 days vs. 113 +/- 36 days, p = 0.006). There was one case of exit site infection in a patient with hepatic hydrothorax. Among subjects who were alive at 3 months after the catheter removal, none had recurrence of their pleural effusion. CONCLUSION: The Denver catheter was effective in achieving pleurodesis in non-malignant pleural effusions. The complication rate was low and patient satisfaction was high. PMID- 21636654 TI - Interpeduncular heterotopia in Joubert syndrome: a previously undescribed MR finding. AB - The so-called molar tooth sign is the radiologic hallmark of JSRD. Joubert syndrome is a rare, most often autosomal-recessive disorder with a characteristic malformation of the midhindbrain. We describe 3 patients with JSRD and the additional MR finding of tissue resembling heterotopia in the interpeduncular fossa, which in one patient was combined with a more extensive intramesencephalic heterotopia. Interpeduncular heterotopia has not been reported previously, either in the context of JSRD or as a separate entity. This new imaging feature enlarges the spectrum of brain stem abnormalities in JSRD. In view of the underlying ciliopathy, it seems likely that the interpeduncular heterotopia results from misdirected migration. PMID- 21636655 TI - Lorcaserin, a 5-HT2C agonist, decreases nicotine self-administration in female rats. AB - Lorcaserin, a selective 5-hydroxytryptamine(2C) (5-HT(2C)) agonist, has been shown to facilitate weight loss in obese populations. It was assessed for its efficacy in reducing nicotine self-administration in young adult female Sprague Dawley rats. The effect of short-term doses (subcutaneous) on nicotine self administration (0.03 mg/kg per infusion) with a fixed ratio 1 schedule was assessed in 3-h sessions. Short-term lorcaserin doses (0.3125-20 mg/kg) were administered in a counterbalanced order. Significant reduction of nicotine self administration was achieved with all of the short-term doses in this range. Tests of lorcaserin on locomotor activity detected prominent sedative effects at doses greater than 1.25 mg/kg with more modest transient effects seen at 0.625 to 1.25 mg/kg. Long-term effects of lorcaserin on locomotor activity were tested with repeated injections with 0.625 mg/kg lorcaserin 10 times over 2 weeks. This low lorcaserin dose did not cause an overall change in locomotor activity relative to that of saline-injected controls. Long-term lorcaserin (0.625 mg/kg) significantly reduced nicotine self-administration over a 2-week period of repeated injections. Long-term lorcaserin at this same dose had no significant effects on food self-administration over the same 2-week period of repeated injections. These studies support development of the 5-HT(2C) agonist lorcaserin to aid tobacco smoking cessation. PMID- 21636656 TI - Subchronic administration and combination metabotropic glutamate and GABAB receptor drug therapy in fragile X syndrome. AB - The most common cause of inherited mental retardation, fragile X syndrome, results from a triplet repeat expansion in the FMR1 gene and loss of the mRNA binding protein, fragile X mental retardation protein (FMRP). In the absence of FMRP, signaling through group I metabotropic glutamate receptors (mGluRs) is enhanced. We previously proposed a mechanism whereby the audiogenic seizures exhibited by FMR1 null mice result from an imbalance in excitatory mGluR and inhibitory GABA(B) receptor (GABA(B)R) signaling (Mol Pharmacol 76:18-24, 2009). Here, we tested the mGluR5-positive allosteric modulator 3-cyano-N-(1,3-diphenyl 1H-pyrazol-5-yl)benzamide (CDPPB), the mGluR5 inverse agonist 2-methyl-6 (phenylethynyl)pyridine (MPEP), and GABA(B) receptor agonists, alone and in combination on receptor protein expression and audiogenic seizures in FMR1 mice. Single doses of MPEP (30 mg/kg), the GABA(B)R orthosteric agonist R-baclofen (1 mg/kg), or the GABA(B)R-positive allosteric modulator N,N'-dicyclopentyl-2 (methylthio)-5-nitro-4,6-pyrimidine diamine (GS-39783) (30 mg/kg), reduced the incidence of seizures. However, when administered subchronically (daily injections for 6 days), MPEP retained its anticonvulsant activity, whereas R baclofen and GS-39783 did not. When administered at lower doses that had no effect when given alone, a single injection of MPEP plus R-baclofen also reduced seizures, but the effect was lost after subchronic administration. We were surprised to find that subchronic treatment with R-baclofen also induced tolerance to a single high dose of MPEP. These data demonstrate that tolerance develops rapidly to the antiseizure properties of R-baclofen alone and R-baclofen coadministered with MPEP, but not with MPEP alone. Our findings suggest that cross-talk between the G-protein signaling pathways of these receptors affects drug efficacy after repeated treatment. PMID- 21636657 TI - Navigating a tool end in a specific direction: stick-tool use in kea (Nestor notabilis). AB - This study depicts how captive kea, New Zealand parrots, which are not known to use tools in the wild, employ a stick-tool to retrieve a food reward after receiving demonstration trials. Four out of six animals succeeded in doing so despite physical (beak curvature) and ecological (no stick-like materials used during nest construction) constraints when handling elongated objects. We further demonstrate that the same animals can thereafter direct the functional end of a stick-tool into a desired direction, aiming at a positive option while avoiding a negative one. PMID- 21636658 TI - The association of traumatic experiences and posttraumatic stress disorder with physical morbidity in old age: a German population-based study. AB - OBJECTIVE: To examine the relationship of traumatic experiences and posttraumatic stress disorder (PTSD) among a representative population sample of 1456 German elderly (60-85 years). Several studies have suggested that PTSD and traumatic experiences are related to adverse health outcomes. However, many past studies were based on special samples such as combat veterans or survivors of natural disasters. METHODS: Using self-report data and regression analyses, we investigated the association of traumatic experiences and PTSD with several medical conditions. RESULTS: Traumatized subjects had a significantly increased risk for all the medical conditions under study compared with those participants without a traumatic exposure, ranging from odds ratio of 1.37 (95% confidence interval [CI] = 1.07-1.75) for hypertension up to 5.12 (95% CI = 2.25-11.6) for cancer. There are significant associations of current PTSD with cardiovascular diseases (angina pectoris/coronary artery disease, congestive heart failure, and peripheral vascular disease) and cardiovascular risk factors (hypertension and elevated cholesterol level; odds ratio of 1.94 [95% CI = 1.14-3.31]) for peripheral vascular disease up to 3.76 [95% CI = 2.11-6.70] for elevated cholesterol level), as well as with asthma, cancer, back pain, hard of hearing, osteoporosis, stomach problems, and thyroid disorders. CONCLUSIONS: These findings suggest an association between traumatic stress and PTSD with impaired physical health in a general population sample in the German elderly. It underscores the importance of traumatic experiences and PTSD not only for mental health but also for physical health as a long-term consequence. PMID- 21636659 TI - Directionality of the relationship between depressive symptom dimensions and C reactive protein in patients with acute coronary syndromes. AB - OBJECTIVE: Previous theoretical models predict that elevated inflammation may predict later depressive symptoms, but bidirectional associations are possible. We examined whether depressive symptoms or inflammation predicts change in the other for a 3-month period in a sample of adults with acute coronary syndromes (ACS). METHODS: During hospitalization for their index ACS event (baseline) and then again 1 and 3 months later, 163 post-ACS patients completed the Beck Depression Inventory, a measure of depressive symptom severity with cognitive affective and somatic-affective subscales. C-reactive protein (CRP) was also assessed at each visit; known correlates of depression and CRP were assessed at baseline. Path analyses were conducted to evaluate prospective associations between depressive symptoms and log-transformed CRP values and whether strength and/or directionality varied by specific depressive symptom dimensions. RESULTS: Baseline total depressive symptom severity predicted a smaller decrease in CRP from baseline to 1 month (unstandardized parameter estimates [B] = 0.04, p < .001) controlling for all covariates, as did baseline cognitive-affective depressive symptom severity (B = 0.10, p = .02). Baseline somatic-affective depressive symptom severity did not predict change in CRP (B = -0.002, p = .94). CRP did not predict 1- or 3-month change in total, cognitive-affective, or somatic-affective depressive symptom severity. The results did not differ for men and women. CONCLUSIONS: Greater cognitive-affective and total depressive symptom severity at the time of a cardiac event predicts a smaller decrease in CRP 1 month later, but there was no evidence in this study that CRP predicts change in depressive symptoms. PMID- 21636660 TI - Depressed mood and flow-mediated dilation: a systematic review and meta-analysis. AB - OBJECTIVE: This systematic and quantitative review evaluates the literature on associations between depressed mood and flow-mediated dilation (FMD), a measure of endothelial function, in adults. METHODS: Published English-language articles (through December 2010) were identified from literature searches, assessed for data extraction, and evaluated for quality. RESULTS: The literature includes cross-sectional (n = 9) and retrospective examinations (n = 3) of how FMD correlates with clinical or subclinical depression in healthy adults and cardiovascular patients (total N across 12 studies = 1491). FMD was assessed using a variety of methodologies. Samples were predominately older white and Asian subjects with higher socioeconomic status. In eight of the 12 articles selected for this review, at least one significant inverse association was noted between depressed mood and FMD, with primarily moderate effect sizes. The overall meta-analysis (random-effects model) revealed a combined effect size of correlation coefficient r = 0.19 (95% confidence interval = 0.08-0.29, p = .001). Significant combined effects were found for subgroups of studies that a) received better quality ratings (r = 0.29), b) examined patients with cardiovascular disease or with cardiovascular disease risk factors/comorbidity (r = 0.29), c) used maximum vasodilation to quantify FMD (r = 0.27), and d) assessed samples that had a mean age of 55 years and older (r = 0.15). CONCLUSIONS: Diverse studies support the inverse correlation between depressed mood and endothelial function, as measured by FMD. This literature would be strengthened by prospective studies, increased methodological consistency in FMD testing, and broader sampling (e.g., African Americans, younger age, lower socioeconomic status). PMID- 21636661 TI - Evaluation of enzyme immunoassay and radioimmunoassay methods for the measurement of plasma oxytocin. AB - OBJECTIVE: There is increased interest in measuring peripheral oxytocin levels to better understand the role of this peptide in mammalian behavior, physiology, and disease. The purpose of this study was to compare methods for plasma oxytocin measurement using a commercially available enzyme immunoassay (EIA) and radioimmunoassay (RIA), to evaluate the need for sample extraction, and to assess the immunospecificity of the assays. METHODS: Oxytocin was measured in extracted and unextracted human plasma samples (n = 39). Oxytocin and its degradation products were separated by high-performance liquid chromatography and gel filtration chromatography and then assayed by EIA or RIA to identify oxytocin immunoreactive peaks. RESULTS: Without extraction, plasma measured by EIA was more than 100-fold higher than in extracted plasma, and the correlation between oxytocin levels in extracted and unextracted plasma was minimal (Spearman rho = 0.10, p = .54). Using the RIA, most samples (>90%) were below the level of detection with or without extraction. After chromatographic fractionation of sample extracts, multiple immunoreactive products were found to be present in addition to oxytocin, which casts doubts on the specificity of the assays. CONCLUSIONS: Changes in oxytocin levels have been reported in social and behavioral challenge studies. This study indicates that sample extraction is necessary to obtain valid assay results. Changes in oxytocin degradation products are likely to contribute to the previously observed responses in circulating oxytocin levels to behavioral and social challenge. There is a critical need for valid and reliable methods to measure oxytocin in biologic samples. PMID- 21636662 TI - Temporal uncoupling of the DNA methylome and transcriptional repression during embryogenesis. AB - DNA methylation is a tightly regulated epigenetic mark associated with transcriptional repression. Next-generation sequencing of purified methylated DNA obtained from early Xenopus tropicalis embryos demonstrates that this genome is heavily methylated during blastula and gastrula stages. Although DNA methylation is largely absent from transcriptional start sites marked with histone H3 lysine 4 trimethylation (H3K4me3), we find both promoters and gene bodies of active genes robustly methylated. In contrast, DNA methylation is absent in large H3K27me3 domains, indicating that these two repression pathways have different roles. Comparison with chromatin state maps of human ES cells reveals strong conservation of epigenetic makeup and gene regulation between the two systems. Strikingly, genes that are highly expressed in pluripotent cells and in Xenopus embryos but not in differentiated cells exhibit relatively high DNA methylation. Therefore, we tested the repressive potential of DNA methylation using transient and transgenic approaches and show that methylated promoters are robustly transcribed in blastula- and gastrula-stage embryos, but not in oocytes or late embryos. These findings have implications for reprogramming and the epigenetic regulation of pluripotency and differentiation and suggest a relatively open, pliable chromatin state in early embryos followed by reestablished methylation dependent transcriptional repression during organogenesis and differentiation. PMID- 21636663 TI - Using endobronchial ultrasound features to predict lymph node metastasis in patients with lung cancer. AB - PURPOSES: Reliable staging of the mediastinum determines TNM classification and directs therapy for non-small cell lung cancer (NSCLC). Our aim was to evaluate predictors of mediastinal lymph node metastasis in patients undergoing endobronchial ultrasound (EBUS). METHODS: Patients with known or suspected lung cancer undergoing EBUS for staging were included. Lymph node radiographic characteristics on chest CT/PET scan and ultrasound characteristics of size, shape, border, echogenicity, and number were correlated with rapid on-site evaluation (ROSE) and final pathology. Logistic regression (estimated with generalized estimating equations to account for correlation across nodes within patients) was used with cancer (vs normal pathology) as the outcome. ORs compare risks across groups, and testing was performed with two-sided alpha of 0.05. RESULTS: Two hundred twenty-seven distinct lymph nodes (22.5% positive for malignancy) were evaluated in 100 patients. Lymph node size, by CT scan and EBUS measurements, and round and oval shape were predictive of mediastinal metastasis. Increasing size of lymph nodes on EBUS was associated with increasing malignancy risk (P = .0002). When adjusted for CT scan size, hypermetabolic lymph nodes on PET scan did not predict malignancy. Echogenicity and border contour on EBUS and site of biopsy were not significantly associated with cancer. In 94.8% of lymph nodes with a clear diagnosis, the ROSE of the first pass correlated with subsequent passes. CONCLUSIONS: Lymph node size on CT scan and EBUS and round or oval shape by EBUS are predictors of malignancy, but no single characteristic can exclude a visualized lymph node from biopsy. Further, increasing the number of samples taken is unlikely to significantly improve sensitivity. PMID- 21636664 TI - Preventive and therapeutic effects of phosphoinositide 3-kinase inhibitors on acute lung injury. AB - BACKGROUND: Phosphoinositide 3-kinases (PI3Ks) are involved in a number of biologic responses. Recent preclinical studies demonstrated that the PI3K dominant signal pathway could play an important role in the development of acute lung injury, although the mechanism remains unclear. METHODS: CD-1 mice were administered different PI3K inhibitors either intranasally or intragastrically once a day for 3 days before intratracheal instillation of lipopolysaccharide at 4 h and 24 h. Effects of SHBM1009 on lipopolysaccharide-induced capillary permeability, leukocyte distribution and activation, and epithelial cell function were measured. Therapeutic effects of SHBM1009 on pancreatic elastase-induced lung injury were evaluated in rats. RESULTS: The data demonstrated that the local delivery of PI3K inhibitors played more effective roles in the prevention of endotoxin-induced lung injury than the systemic delivery. The preventive effects of PI3K inhibitors varied most likely because of chemical properties, targeting sites, and pharmacokinetics. The local PI3K inhibitors prevented both endotoxin- and elastase-induced lung injury in mice and rats, possibly through directly inhibiting or inactivating the function of airway epithelial cells, which could not produce chemoattractant factors to activate neutrophils and macrophages. CONCLUSIONS: PI3K may be a therapeutic target for lung injury, and local delivery of PI3K inhibitors may be one of the optimal approaches for the therapy. PMID- 21636665 TI - Racial differences in quality of life in patients with COPD. AB - BACKGROUND: Although COPD is associated with significant health-related quality of-life (HRQL) impairment, factors influencing HRQL in patients with COPD are not well understood, particularly in African Americans. We hypothesized that HRQL in COPD differs by race and sought to identify factors associated with those differences. METHODS: We analyzed 224 African American and 1,049 Caucasian subjects with COPD enrolled in the COPDGene (Genetic Epidemiology of COPD) Study whose conditions were classified as GOLD (Global Initiative for Chronic Obstructive Lung Disease) stages I to IV. HRQL and symptoms were compared using the St. George Respiratory Questionnaire (SGRQ) and the modified Medical Research Council Dyspnea (MMRC) scale. We constructed a mixed-effects linear regression model for SGRQ score. RESULTS: African Americans were younger and reported fewer pack-years of smoking, more current smoking, and less attained education than Caucasians; MMRC scores were higher (P = .02) as were SGRQ scores (mean score difference, 8.4; P < .001). In a general linear model of SGRQ total score after adjusting for factors such as age, sex, and pack-years of smoking, SGRQ total score was similar for African Americans and Caucasians who reported no COPD exacerbations in the prior year. However, for subjects with exacerbations, SGRQ total score was increased to a greater relative extent for African Americans than for Caucasians (1.89 points for each exacerbation, P = .006). For hospitalized exacerbations, the effect on SGRQ total score also was greater for African Americans (4.19 points, P = .04). Furthermore, a larger percentage of African Americans reported having had at least one exacerbation that required hospitalization in the prior year (32% vs 16%, P < .001). CONCLUSION: In analyses that account for other variables that affect quality of life, HRQL is similar for African Americans and Caucasians with COPD without exacerbations but worse for African Americans who experience exacerbations, particularly hospitalized exacerbations. PMID- 21636666 TI - Efficacy of an adjustable oral appliance and comparison with continuous positive airway pressure for the treatment of obstructive sleep apnea syndrome. AB - BACKGROUND: We sought to establish the efficacy of an adjustable oral appliance (aOA) in the largest patient population studied to date, to our knowledge, and to provide a comparison with continuous positive airway pressure (CPAP). METHODS: We conducted a retrospective analysis of patients using an aOA. Results of overnight polysomnography with aOA titration were evaluated and compared with CPAP. Predictors of a successful aOA titration were determined using a multivariate logistic regression model. RESULTS: A total of 497 patients were given an aOA during the specified time period. The aOA reduced the mean apnea-hypopnea index (AHI) to 8.4 +/- 11.4, and 70.3%, 47.6%, and 41.4% of patients with mild, moderate, and severe disease achieved an AHI < 5, respectively. Patients using an aOA decreased their mean Epworth Sleepiness Score by 2.71 (95% CI, 2.3-3.2; P < .001) at follow-up. CPAP improved the AHI by -3.43 (95% CI, 1.88-4.99; P < .001) when compared with an aOA, but when adjusted for severity of disease, this difference only reached significance for patients with severe disease (-5.88 [95% CI, -8.95 to -2.82; P < .001]). However, 70.1% of all patients achieved an AHI < 5 using CPAP compared with 51.6% for the aOA (P < .001). On multivariate analysis, baseline AHI was a significant predictor of achieving an AHI < 5 on aOA titration, and age showed a trend toward significance. CONCLUSIONS: In comparison with past reports, more patients in our study achieved an AHI < 5 using an aOA. The aOA is comparable to CPAP for patients with mild disease, whereas CPAP is superior for patients with moderate to severe disease. A lower AHI was the only predictor of a successful aOA titration. PMID- 21636667 TI - Fluticasone/Salmeterol combination confers benefits in people with asthma who smoke. AB - BACKGROUND: Smoking induces airway inflammation and relative resistance to inhaled steroids. The objective of this study was to evaluate the effects on airway hyperresponsiveness of adding salmeterol to fluticasone vs doubling the dose of fluticasone in patients with asthma who smoked and patients with asthma who did not smoke. METHODS: Sixteen patients with mild to moderate persistent asthma who did not smoke and 15 such patients who smoked completed a double blind, randomized, placebo-controlled crossover study. They received either a fluticasone/salmeterol combination (FP/SM) (125/25 MUg) two puffs bid (plus fluticasone placebo), or active fluticasone (250 MUg) two puffs bid (plus FP/SM placebo), for 2 weeks each, with baselines after 1-week to 2-week run-in and washout periods. The primary outcome was the change from baseline in the provocative concentration of methacholine required to produce a 20% fall in FEV(1) (PC(20)). RESULTS: In the patients who did not smoke, there were similar improvements in the methacholine PC(20) with the use of fluticasone and FP/SM. The patients who smoked gained a benefit from FP/SM but not fluticasone, amounting to a PC(20) difference of 1.6 doubling dilutions (95% CI, 1.0-2.2), P < .01. The provocative dose of mannitol required to produce a 15% fall in FEV(1) (PD(15)) showed greater improvements with FP/SM than fluticasone in both patients who smoked and did not smoke. Similar differences in airway caliber between those who smoked and did not smoke were observed in FEV(1) and airway resistance. CONCLUSIONS: FP/SM confers greater improvements in airway hyperresponsiveness and airway caliber in patients with asthma who smoke compared with double the dose of fluticasone. We hypothesize that in the presence of relative steroid resistance, the smooth muscle stabilization conferred by salmeterol is of greater clinical importance in patients who smoke than in those who do not smoke. TRIAL REGISTRY: ClinicalTrials.gov: No.: NCT00830505; URL: www.clinicaltrials.gov. PMID- 21636668 TI - Relation of natriuretic peptide concentrations to central sleep apnea in patients with heart failure. AB - BACKGROUND: Central sleep apnea (CSA) is frequent among patients with heart failure (HF) and associated with increased morbidity and mortality. Elevated cardiac filling pressures promote CSA and atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) secretion. We hypothesized that circulating natriuretic peptide concentrations predict CSA. METHODS: Consecutive patients with HF (n = 44) with left ventricular ejection fraction (LVEF) <= 35% underwent polysomnography for detection of CSA. CSA was defined as an apnea-hypopnea index >= 15 with >= 50% central apneic events. The relation of natriuretic peptide concentrations to CSA was evaluated by estimation of ORs and receiver operator characteristics (ROCs). RESULTS: Twenty-seven subjects (61%) had CSA, with men more frequently affected than women (73% vs 27%; OR, 7.1; P = .01); given that only three women had CSA, further analysis was restricted to men. Subjects with CSA had higher mean ANP (4,336 pg/mL vs 2,510 pg/mL, P = .03) and BNP concentrations (746 pg/mL vs 379 pg/mL, P = .05). ANP and BNP concentrations were significantly related to CSA (OR, 3.7 per 3,000 pg/mL, P = .03 and OR, 1.5 per 200 pg/mL, P = .04, respectively), whereas age, LVEF, and New York Heart Association functional class were not. Concentrations of ANP and BNP were predictive of CSA as ROC demonstrated areas under the curve of 0.75 and 0.73, respectively. CONCLUSIONS: Risk of CSA is related to severity of HF. ANP and BNP concentrations performed similarly for detection of CSA; low concentrations appear associated with low risk for CSA in men. PMID- 21636669 TI - Novel pathway of metabolic activation of bisphenol A-related compounds for estrogenic activity. AB - We previously demonstrated that estrogenic activity of bisphenol A (BPA) in the yeast estrogen screening assay was increased severalfold after incubation with rat liver S9 fraction in the presence of a NADPH-generating system. In this study, we investigated whether eight BPA-related compounds are similarly activated metabolically by rat liver S9 fraction. Three of the analogs exhibited an increase of estrogenic activity after incubation with rat liver S9 fraction but not with microsomal or cytosolic fraction alone. The structures of the metabolites formed were examined by liquid chromatography/mass spectrometry. In addition to oxidized metabolites such as catechols, we found novel dimer-type metabolites. Some of the putative metabolites were chemically synthesized to confirm their structures. The structural requirements for formation of the metabolites, some of which showed more potent estrogenic activity than the parent substrates, were examined. We have uncovered a new pathway of metabolic activation of certain phenolic compounds, such as BPA analogs, to estrogenic dimer-type compounds. PMID- 21636670 TI - Plant polyphenolic compounds as potential antimicrobial drugs. PMID- 21636671 TI - Protease inhibitors decrease IgG shedding from Staphylococcus aureus, increasing complement activation and phagocytosis efficiency. AB - Staphylococcus aureus is a major pathogen for immunologically intact humans and its pathogenesis is a model system for evasion of host defences. Antibodies and complement are essential elements of the humoral immune system for prevention and control of S. aureus infections. The specific hypothesis for the proposed research is that S. aureus modifies humoral host defences by cleaving IgG that has bound to the bacterial surface, thereby inhibiting opsonophagocytosis. S. aureus was coated with pooled, purified human IgG and assayed for the shedding of cleaved IgG fragments using ELISA and Western blot analysis. Surface-bound IgG was shed efficiently from S. aureus in the absence of host blood proteins. Broad spectrum protease inhibitors prevented cleavage of IgG from the S. aureus surface, suggesting that staphylococcal proteases are responsible for IgG cleavage. Serine protease inhibitors and cysteine protease inhibitors decreased the cleavage of surface-bound IgG; however, a metalloprotease inhibitor had no effect. Using protease inhibitors to prevent the cleavage of surface-bound IgG increased the binding of complement C3 fragments on the surface of S. aureus, increased the association with human neutrophils and increased phagocytosis by human neutrophils. PMID- 21636673 TI - Serotype and genotype distributions of pneumococcal carriage isolates recovered from Brazilian children attending day-care centres. AB - Pneumococcal nasopharyngeal carriage isolates recovered from Brazilian children attending day-care centres in 2005 were assessed for serotype, genotype and penicillin susceptibility phenotype. As 124 of the 253 isolates (49 %) were characterized previously with respect to serotype and penicillin susceptibility, the primary objectives were to examine clonal associations and penicillin susceptibility within major serotypes and to assess the suitability of conventional multiplex PCR for deducing carriage serotypes within this population. Using a combination of PCR-based serotyping and the Quellung reaction, serotypes were identified for 81 % (205/253) of the isolates, with serogroups or types 14, 6, 23F, 19F and 18 being predominant. Included within the 205 isolates successfully serotyped by PCR were 28 isolates that had become non viable. Forty-eight isolates were non-typable using both the PCR method and the Quellung reaction. Penicillin non-susceptibility was observed within 16 of the 18 multilocus sequence types detected. Thus, this study provides further evidence from a diverse collection of pneumococcal clones that PCR-based serotype deduction is useful for providing supportive evidence for pneumococcal conjugate vaccine implementation. PMID- 21636675 TI - Chinese translation and validation of the Walking Impairment Questionnaire in patients with peripheral artery disease. AB - The Walking Impairment Questionnaire (WIQ) is a frequently used questionnaire to evaluate patients with intermittent claudication on four subscales: pain severity, walking distance, walking speed and the ability to climb stairs. The aim of this study is to translate and validate the WIQ in Chinese. After translation and cultural adaptation of the WIQ, 134 patients with intermittent claudication completed the Chinese WIQ and European Quality of Life 5 Dimension (EQ-5D). Walking distances were determined by the 6-minute walk test (6MWT). Correlations between the WIQ, quality of life questionnaire and walking distances were calculated to determine validity. Reliability and internal consistency were determined using the intra-class correlation coefficient (ICC) and Cronbach's alpha (alpha), respectively. Significant correlations were found between the WIQ score, initial claudication distance (ICD), absolute claudication distance (ACD) and all domains of the EQ-5D (all p <= 0.01). Test-retest reliability (ICC = 0.74) and the overall internal consistency determined (alpha = 0.90) showed good agreement. A lower WIQ score corresponded to shorter walking distances. In conclusion, this study showed that the Chinese version of the WIQ is a valid, reliable and clinically relevant instrument for assessing walking impairment in patients with intermittent claudication. PMID- 21636674 TI - MicroRNA processing and binding site polymorphisms are not replicated in the Ovarian Cancer Association Consortium. AB - BACKGROUND: Single nucleotide polymorphisms (SNP) in microRNA-related genes have been associated with epithelial ovarian cancer (EOC) risk in two reports, yet associated alleles may be inconsistent across studies. METHODS: We conducted a pooled analysis of previously identified SNPs by combining genotype data from 3,973 invasive EOC cases and 3,276 controls from the Ovarian Cancer Association Consortium. We also conducted imputation to obtain dense coverage of genes and comparable genotype data for all studies. In total, 226 SNPs within 15 kb of 4 miRNA biogenesis genes (DDX20, DROSHA, GEMIN4, and XPO5) and 23 SNPs located within putative miRNA binding sites of 6 genes (CAV1, COL18A1, E2F2, IL1R1, KRAS, and UGT2A3) were genotyped or imputed and analyzed in the entire dataset. RESULTS: After adjustment for European ancestry, no overall association was observed between any of the analyzed SNPs and EOC risk. CONCLUSIONS: Common variants in these evaluated genes do not seem to be strongly associated with EOC risk. IMPACT: This analysis suggests earlier associations between EOC risk and SNPs in these genes may have been chance findings, possibly confounded by population admixture. To more adequately evaluate the relationship between genetic variants and cancer risk, large sample sizes are needed, adjustment for population stratification should be carried out, and use of imputed SNP data should be considered. PMID- 21636676 TI - Poorer clock draw test scores are associated with greater functional impairment in peripheral artery disease: the Walking and Leg Circulation Study II. AB - We hypothesized that, in the absence of clinically recognized dementia, cognitive dysfunction measured by the clock draw test (CDT) is associated with greater functional impairment in men and women with peripheral artery disease (PAD). Participants were men and women aged 60 years and older with Mini-Mental Status Examination scores >= 24 with PAD (n = 335) and without PAD (n = 234). We evaluated the 6-minute walk test, 4-meter walking velocity at usual and fastest pace, the Short Physical Performance Battery (SPPB), and accelerometer-measured physical activity. CDTs were scored using the Shulman system as follows: Category 1 (worst): CDT score 0-2; Category 2: CDT score 3; Category 3 (best): CDT score 4 5. Results were adjusted for age, sex, race, education, ankle-brachial index (ABI), and comorbidities. In individuals with PAD, lower CDT scores were associated with slower 4-meter usual-paced walking velocity (Category 1: 0.78 meters/second; Category 2: 0.83 meters/second; Category 3: 0.86 meters/second; p trend = 0.025) and lower physical activity (Category 1: 420 activity units; Category 2: 677 activity units; Category 3: 701 activity units; p-trend = 0.045). Poorer CDT scores were also associated with worse functional performance in individuals without PAD (usual and fast-paced walking velocity and SPPB, p-trend = 0.022, 0.043, and 0.031, respectively). In conclusion, cognitive impairment identified with CDT is independently associated with greater functional impairment in older, dementia-free individuals with and without PAD. Longitudinal studies are necessary to explore whether baseline CDT scores and changes in CDT scores over time can predict long-term decline in functional performance in individuals with and without PAD. PMID- 21636677 TI - Effect of darbepoetin alfa on endothelial progenitor cells and vascular reactivity in chronic kidney disease. AB - Endothelial progenitor cells (EPCs) are thought to be important for maintaining normal vascular function. We conducted a prospective study evaluating the effect of the erythropoiesis-stimulating agent darbepoetin alfa on EPCs and vascular function in patients with chronic kidney disease (CKD), with or without diabetes. Thirty subjects with CKD (20 subjects with type II diabetes mellitus and 10 without diabetes mellitus) received weekly subcutaneous administration of darbepoetin alfa for 4 weeks. EPCs were measured at baseline and 2 and 4 weeks after drug administration. Vascular function was measured with brachial ultrasound and cell activity was measured with a cell proliferation assay. Cells expressing CD133, CD34, CD146 and CD146/31 were significantly elevated (all p < 0.05), flow-mediated vasodilatation increased 2.1%, 95% CI: (0.4%, 3.8%) and colony-forming units increased twofold, 95% CI: (1.7, 2.3) after 4 weeks of treatment with darbepoetin alfa. Subjects with diabetes exhibited an increase in a subset of EPCs (CD133( +) and 34(+), p < 0.01 and p = 0.06, respectively), vasodilatation and proliferation. In conclusion, the administration of darbepoetin alfa for 4 weeks increased a subset of EPCs, improved endothelial function and increased cell proliferation, including those with diabetes, which is consistent with a favorable improvement in vascular health. PMID- 21636678 TI - Images in vascular medicine: perforator vein incompetence in a 10-year-old male. PMID- 21636679 TI - Images in vascular medicine: rapid epinephrine 'reversal' with phentolamine following accidental autoinjector inoculation. PMID- 21636680 TI - Differential secretomics of Streptococcus pyogenes reveals a novel peroxide regulator (PerR)-regulated extracellular virulence factor mitogen factor 3 (MF3). AB - Streptococcus pyogenes is a human pathogen that causes various diseases. Numerous virulence factors secreted by S. pyogenes are involved in pathogenesis. The peroxide regulator (PerR) is associated with the peroxide resistance response and pathogenesis, but little is known about the regulation of the secretome involved in virulence. To investigate how PerR regulates the expression of the S. pyogenes secretome involved in virulence, a perR deficient mutant was used for comparative secretomic analysis with a wild-type strain. The conditioned medium containing secreted proteins of a wild-type strain and a perR deficient mutant at the stationary phase were collected for two-dimensional gel electrophoresis analysis, where protease inhibitors were applied to avoid the degradation of extracellular proteins. Differentially expressed protein spots were identified by liquid chromatography electrospray ionization tandem MS. More than 330 protein spots were detected on each gel. We identified 25 unique up-regulated proteins and 13 unique down-regulated proteins that were directly or indirectly controlled by the PerR regulator. Among these identified proteins, mitogen factor 3 (MF3), was selected to verify virulence and the expression of gene products. The data showed that MF3 protein levels in conditioned medium, as measured by immunoblot analysis, correlated well with protein levels determined by two-dimensional gel electrophoresis analysis. We also demonstrated that PerR bound to the promoter region of the mf3 gene. The result of an infection model showed that virulence was attenuated in the mf3 deficient mutant. Additional growth data of the wild type strain and the mf3 deficient mutant suggested that MF3 played a role in digestion of exogenous DNA for promoting growth. To summarize, we conclude that PerR can positively regulate the expression of the secreted protein MF3 that contributes to the virulence in S. pyogenes. The analysis of the PerR-regulated secretome provided key information for the elucidation of the host-pathogen interactions and might assist in the development of potential chemotherapeutic strategies to prevent or treat streptococcal diseases. PMID- 21636681 TI - HSP70 inhibition by the small-molecule 2-phenylethynesulfonamide impairs protein clearance pathways in tumor cells. AB - The evolutionarily conserved stress-inducible HSP70 molecular chaperone plays a central role in maintaining protein quality control in response to various forms of stress. Constitutively elevated HSP70 expression is a characteristic of many tumor cells and contributes to their survival. We recently identified the small molecule 2-phenylethyenesulfonamide (PES) as a novel HSP70 inhibitor. Here, we present evidence that PES-mediated inhibition of HSP70 family proteins in tumor cells results in an impairment of the two major protein degradation systems, namely, the autophagy-lysosome system and the proteasome pathway. HSP70 family proteins work closely with the HSP90 molecular chaperone to maintain the stability and activities of their many client proteins, and PES causes a disruption in the HSP70/HSP90 chaperone system. As a consequence, many cellular proteins, including known HSP70/HSP90 substrates, accumulate in detergent insoluble cell fractions, indicative of aggregation and functional inactivation. Overall, PES simultaneously disrupts several cancer critical survival pathways, supporting the idea of targeting HSP70 as a potential approach for cancer therapeutics. PMID- 21636682 TI - The ARF tumor suppressor inhibits tumor cell colonization independent of p53 in a novel mouse model of pancreatic ductal adenocarcinoma metastasis. AB - Pancreatic ductal adenocarcinoma (PDAC) is an incurable, highly metastatic disease that is largely resistant to existing treatments. A better understanding of the genetic basis of PDAC metastasis should facilitate development of improved therapies. To that end, we developed a novel mouse xenograft model of PDAC metastasis to expedite testing of candidate genes associated with the disease. Human PDAC cell lines BxPC-3, MiaPaCa-2, and Panc-1 stably expressing luciferase were generated and introduced by intracardiac injections into immunodeficient mice to model hematogenous dissemination of cancer cells. Tumor development was monitored by bioluminescence imaging. Bioluminescent MiaPaCa-2 cells most effectively recapitulated PDAC tumor development and metastatic distribution in vivo. Tumors formed in nearly 90% of mice and in multiple tissues, including normal sites of PDAC metastasis. Effects of p14ARF, a known suppressor of PDAC, were tested to validate the model. In vitro, p14ARF acted through a CtBP2 dependent, p53-independent pathway to inhibit MiaPaCa-2-invasive phenotypes, which correlated with reduced tumor cell colonization in vivo. These findings establish a new bioluminescent mouse tumor model for rapidly assessing the biological significance of suspected PDAC metastasis genes. This system may also provide a valuable platform for testing innovative therapies. PMID- 21636683 TI - Hypoglycemic effect of basil (Ocimum basilicum) aqueous extract is mediated through inhibition of alpha-glucosidase and alpha-amylase activities: an in vitro study. AB - The present study investigated the in vitro hypoglycemic activity of basil (Ocimum basilicum) aqueous extract. Preliminary phytochemical screening of the extract revealed the presence of reducing sugars, cardiac glycosides, tannins, saponins, glycosides, flavonoids and steroids. The total polyphenols content (TPC), flavonoids content (FC), percentage diphenylpicrylhydrazyl (DPPH( . )) radical inhibition and total antioxidant status (TAS) were estimated. The FC was 41 +/- 2.2 rutin/g dry extract, the TPC was 146 +/- 5.26 mg catechin/g dry extract and the TAS was 5.12 +/- 0.7 mmol/L. The %DPPH( . ) free radical inhibition was 60%, 54%, 49% and 43%, respectively, for different extract concentrations; 20, 18.2, 16.3 and 14.5 mg/ml, respectively. The extract elicited significant dose-dependent pattern against rat intestinal sucrase (RIS; IC(50) = 36.72 mg/ml), rat intestinal maltase (RIM; IC(50) = 21.31 mg/ml) and porcine pancreatic alpha-amylase (PPA; IC(50) = 42.50 mg/ml) inhibitory activities. The inhibition was greater against maltase compared with sucrase. These effects may be attributed to the high TPC and FC levels. The linear regression analysis revealed strong significant positive correlations between %DPPH( . ) radical inhibition and each of %RIS, %RIM and %PPA inhibiting activity. Also, strong significant positive correlations between %RIS and either %RIM or %PPA inhibition activity were observed. We concluded therefore that basil aqueous extract via antioxidant and possibly alpha-glucosidase and alpha-amylase inhibiting activities, offered positive benefits to control diabetes. PMID- 21636684 TI - Effect of nickel and chromium exposure on buccal cells of electroplaters. AB - The electroplating industry commonly involves the use of nickel and chromium. An assessment of the genotoxic effects of these metals can be carried out by micronucleus (MN) test in buccal cells. Other nuclear anomalies (NA) observed in buccal cells viz., karyorrhexis, pyknosis and karyolysis are also the indicators of genotoxicity. The current study aims at determining the extent of genotoxic damage in relation to the duration of exposure to nickel and hexavalent chromium via micronuclei induction and other nuclear anomalies. The present investigation included 150 subjects of which 50 individuals with no history of nickel/chromium exposure (Group I) were taken as control, 50 electroplaters exposed to nickel and hexavalent chromium for duration of less than 10 years (Group II) and 50 electroplaters exposed for >=10 years (Group III) were included. Slides of buccal cells were prepared and the frequency of MN (0/00) and NA (0/00) were calculated. ANOVA was applied to test significance. Results were considered significant at p < 0.05 and p < 0.001. Group III showed the highest MN frequency (1.08 +/- 0.540/00, p < 0.05), karyorrhexis (20.75 +/- 6.29, p < 0.05), karyolysis (3.50 +/ 1.91, p < 0.001), binucleate (4.75 +/- 2.75, p < 0.05) and enucleated cells (5.75 +/- 1.70, p < 0.05). Significant increase in frequencies between Group II and III was found as duration of exposure increased. Plasma nickel and chromium levels were also determined which showed a positive correlation with frequency MN and other nuclear abnormalities (p < 0.01). PMID- 21636685 TI - Influence of naringin on cadmium-induced genomic damage in human lymphocytes in vitro. AB - Cadmium is an important toxic environmental heavy metal. Generally, occupational and environmental exposures to cadmium result from heavy metal mining, metallurgy and industrial use and the manufacturing of nickel-cadmium batteries, pigments and plastic stabilizers. Cadmium induces oxidative stress and alters the antioxidant system, resulting in oxidative DNA damage and lipid peroxidation. The effect of naringin, a grapefruit flavonone, on cadmium-induced genomic damage was studied by using an in vitro system to test for chromosomal aberrations and sister chromatid exchanges. Cadmium significantly increased the total chromosomal aberrations in human lymphocytes at concentrations of 20 and 40 MUM, and although naringin alone did not induce any chromosomal aberrations, it decreased those induced by cadmium. The mitotic index was not affected by either cadmium or naringin. Cadmium also induced a significant number of sister chromatid exchanges, but naringin alone did not induce sister chromatid exchanges and was unable to decrease the frequency of sister chromatid exchanges induced by cadmium. Replicative index analysis revealed that naringin and cadmium did not significantly alter replicative index frequencies. In this study, we show that plant-based flavonoids, such as naringin, may reduce the genomic damage induced by cadmium and may protect the cellular environments from free radical damage by its possible antioxidative potential. PMID- 21636686 TI - Improving the access of young urban children to speech, language and hearing screening via telehealth. AB - We studied the feasibility of low-cost videoconferencing (using Skype) in urban community health clinics for speech, language and hearing screening of children up to six years of age. During a two-year study, screening services were provided via videoconferencing at two community clinics in an inner city area of Cleveland, Ohio. In total, 411 screenings were completed. Of these, 358 children (87%) received hearing screenings, 377 (92%) received tympanometry screening and 263 (64%) received speech and language screening only. A total of 151 children were aged three years or under (37%). The reliability of pure tone hearing screening (n = 7), DPOAE screening (n = 51) and speech-language screening (n = 10) was 100%. Typanometry screenings (n = 55) were 84% reliable. Families reported a high level of satisfaction with both the technology and with the videoconferencing. The results indicate that low-cost videoconferencing for screening of speech, language and hearing development in very young children in urban community health clinics is feasible, reliable and strongly supported by the community. PMID- 21636687 TI - A brief self-guided telehealth intervention for post-traumatic stress disorder in combat veterans: a pilot study. AB - To engage more US combat veterans in PTSD treatment, we offered a psychological intervention that could be initiated in primary care and completed using the Internet. Participants (n = 31) were randomized to complete either Written Emotional Disclosure (WED) or time management narratives on a secure Internet website. In the WED group, participants wrote about their thoughts and emotions regarding one traumatic combat experience in three 20-min sessions. Writing instructions encouraged exposure to traumatic memories and cognitive processing of trauma. The intervention was found to be feasible and safe to implement. Although follow-up assessments did not reveal significant group differences in PTSD symptoms, half of the WED participants reported symptom reductions. Content analyses revealed that participants who expressed more emotion and cognitions were significantly more likely to experience decreased PTSD symptoms. WED may have promise as a brief intervention for veterans with PTSD. PMID- 21636688 TI - Tree phenology responses to warming: spring forward, fall back? PMID- 21636689 TI - Temperature signals contribute to the timing of photoperiodic growth cessation and bud set in poplar. AB - Bud set, the cornerstone delimiting the seasonal growth period in trees, is the dynamic net result of the often photoperiod-controlled growth cessation and the subsequent bud formation. Here, we show that in hybrid poplar, the critical day length for growth cessation and the duration of bud formation each vary with local climatic conditions in identical genotypes. The detailed dissection of bud set suggests temperature as one additional environmental factor that modifies the sensitivity to day-length signals at growth cessation and influences the duration of bud formation in poplar. The ability of perennial plants to integrate additional environmental signals with photoperiod signaling may add to short-term acclimatization to the predicted longer growing seasons in future climates. PMID- 21636690 TI - Phenotypic plasticity of stem water potential correlates with crop load in horticultural trees. AB - Conceptual models accounting for the influence of source:sink ratio on water relations of trees are theoretically relevant from a physiological perspective and practically important for irrigation scheduling. Midday stem water potential of horticultural trees often declines with increasing crop load but the actual response depends on environmental, management and plant factors. Here we advance a quantitative synthesis of the response of stem water potential to crop load from the perspective of phenotypic plasticity, defined as 'the amount by which the expression of individual characteristics of a genotype are changed by different environments'. Data sets of stem water potential for contrasting crop loads were compiled for apple (Malus domestica L. Borkh.), olive (Olea europea L.), peach (Prunus persica L.), pear (Pyrus communis L.) and plum (Prunus domestica L.). Phenotypic plasticity of stem water potential was calculated as the slope of the linear regression between stem water potential for each crop load and the environmental mean of stem water potential across crop loads. Regression lines for trees with different crop load diverged with decreasing environmental mean stem water potential. For the pooled data, plasticity of stem water potential was a linear function of relative crop load. This represents a significant shift in perspective: the effect of crop load on the trait per se (stem water potential) is environmentally contingent, but the effect of crop load on the plasticity of the trait is not. We conclude that research on the effects of crop load on tree water relations would return more robust results if plant traits are considered from the dual perspective of the trait per se and its plasticity. PMID- 21636691 TI - Variability of stomatal conductance in a small and isolated population of silver fir (Abies alba Mill.). AB - We analyzed the response to drought of 420 individuals from eight half-sib families from a small and isolated population of silver fir (Abies alba Mill.): 105 of them were kept in well-watered conditions as control while the remaining 315 were exposed to drought for 27 days. A model describing stomatal behavior derived from Monteith and developed in beech by Leonardi et al. was fitted to experimental transpiration data obtained simply from the difference between two daily pot weighings. The estimated parameters were maximum stomatal conductance, maximum transpiration in well-watered conditions and sensitivity to soil water deficit. The model worked well: convergence for all but four individuals and concordance between experimental and fitted data were good (R(2)=0.86). Inter individual variability for all three estimated parameters was high and two of them (maximum stomatal conductance and sensitivity to soil water deficit) were significantly different among families, suggesting genetic control. Our results validate the simplified method used to evaluate individual stomatal parameters. We also show that in the small and isolated population of our study substantial adaptive variability remains, a crucial prerequisite to endure environmental conditions determined by climatic change foreseen for the next decades. PMID- 21636692 TI - Hydraulic responses of whole vines and individual roots of kiwifruit (Actinidia chinensis) following root severance. AB - Whole vine (K(plant)) and individual root (K(root)) hydraulic conductances were measured in kiwifruit (Actinidia chinensis Planch. var. chinensis 'Hort16A') vines to observe hydraulic responses following partial root system excision. Heat dissipation and compensation heat pulse techniques were used to measure sap flow in trunks and individual roots, respectively. Sap flux and measurements of xylem pressure potential (Psi) were used to calculate K(plant) and K(root) in vines with zero and ~80% of roots severed. Whole vine transpiration (E), Psi and K(plant) were significantly reduced within 24 h of root pruning, and did not recover within 6 weeks. Sap flux in intact roots increased within 24 h of root pruning, driven by an increase in the pressure gradient between the soil and canopy and without any change in root hydraulic conductance. Photosynthesis (A) and stomatal conductance (g(s)) were reduced, without significant effects on leaf internal CO(2) concentration (c(i)). Shoot growth rates were maintained; fruit growth and dry matter content were increased following pruning. The woody roots of kiwifruit did not demonstrate a rapid dynamic response to root system damage as has been observed previously in monocot seedlings. Increased sap flux in intact roots with no change in K(root) and only a moderate decline in shoot A suggests that under normal growing conditions root hydraulic conductance greatly exceeds requirements for adequate shoot hydration. PMID- 21636693 TI - Ectomycorrhizal fungal diversity, tree diversity and root nutrient relations in a mixed Central European forest. AB - Knowledge is limited about whether root nutrient concentrations are affected by mixtures of tree species and interspecific root competition. The goal of this field study was to investigate root nutrient element concentrations in relation to root and ectomycorrhizal (EM) diversity in six different mixtures of beech (Fagus sylvatica), ash (Fraxinus excelsior) and lime (Tilia sp.) in an old growth, undisturbed forest ecosystem. Root biomass and nutrient concentrations per tree taxon as well as the abundance and identity of all EM fungi were determined in soil cores of a volume of 1 L (r=40 mm, depth=200 mm). Stand-level nutrient concentrations in overall root biomass and H' (Shannon-Wiener diversity) were obtained by pooling the data per stand. At stand level, Shannon H' for roots and aboveground tree species abundance were correlated. H' for roots and EM fungi were not correlated because of the contribution of ash roots that form only arbuscular mycorrhizal but no EM associations. Nutrient element concentrations in roots showed taxon-related differences and increased in the following order: beech <= lime < ash with the exception of calcium (Ca), which was lower in ash. Stand-level concentrations of Ca, magnesium, potassium and sulfur in roots increased with increasing tree diversity because of two effects: increasing contribution of ash roots to the mixture and increasing Ca accumulation in beech roots with increasing root diversity. On a small scale, increasing root diversity, but not EM diversity, was correlated with decreasing P concentrations in beech roots pointing to interspecific tree competition. Nitrogen (N) concentrations of beech roots were unaltered in relation to root and EM diversity. Opposing behavior was observed for lime and ash: the N concentrations in lime roots increased, whereas those in ash roots decreased with increasing EM diversity in a given soil volume. This suggests that EM diversity facilitates N acquisition of lime roots at the expense of non-EM ash. PMID- 21636694 TI - Initiation of somatic embryogenesis from immature zygotic embryos of oocarpa pine (Pinus oocarpa Schiede ex Schlectendal). AB - The focus of the current project was to establish somatic embryogenesis protocols for the tropical pine species Pinus oocarpa using immature zygotic embryos (ZEs) as explants. Somatic embryogenesis is best supported by mimicking natural seed embryo developmental conditions, through a tissue culture medium formulation based on the mineral content of the seed nutritive tissue [megagametophyte (MG)]. A novel culture medium (P. oocarpa medium, PO) was tested in combination with different plant growth regulator (PGR) concentrations and compared with standard Pinus taeda media for the initiation of somatic embryogenesis from immature ZEs of P. oocarpa. Immature MGs containing immature ZEs of two mother trees were used with 12 and 8% extrusion rates for mother tree genotypes 3 and 5, respectively. In both mother trees the percentage capture was 2%. Multiplication of two captured cell lines (T5C2S01 and T5C1S12) was improved by lowering the concentrations of PGRs to 2.5 uM each 2,4-dichlorophenoxyacetic acid and abscisic acid (ABA) plus 1.0 uM each 6-benzylaminopurine and kinetin. Mature somatic embryos formed on 40 uM ABA, 6% (w/v) maltose, 12% (w/v) PEG 8000 and 0.6% (w/v) Phytagel. While PO medium appeared suboptimal for somatic embryo induction, it did exhibit potential for enhanced culture proliferation and subsequent improved maturation with cell line T5C2S01, where microscopic analysis revealed better embryo morphology on PO medium than on 1250 medium. However, this enhancement was not observed with cell line T5C1S12. Germination was preceded by partial desiccation for a period of 2-3 weeks before transferring the embryos to germination medium. Germination was observed after 7 days under low light, and apical primordia slowly expanded after transfer to ex vitro conditions. To our knowledge, this is the first report on the production of somatic seedlings in P. oocarpa. PMID- 21636695 TI - Normal anatomy and histology of the adult zebrafish. AB - The zebrafish has been shown to be an excellent vertebrate model for studying the roles of specific genes and signaling pathways. The sequencing of its genome and the relative ease with which gene modifications can be performed have led to the creation of numerous human disease models that can be used for testing the potential and the toxicity of new pharmaceutical compounds. Many pharmaceutical companies already use the zebrafish for prescreening purposes. So far, the focus has been on ecotoxicity and the effects on embryonic development, but there is a trend to expand the use of the zebrafish with acute, subchronic, and chronic toxicity studies that are currently still carried out with the more conventional test animals such as rodents. However, before we can fully realize the potential of the zebrafish as an animal model for understanding human development, disease, and toxicology, we must first greatly advance our knowledge of normal zebrafish physiology, anatomy, and histology. To further this knowledge, we describe, in the present article, location and histology of the major zebrafish organ systems with a brief description of their function. PMID- 21636696 TI - The global presence of holistic nursing. PMID- 21636698 TI - Simultaneous phenotyping and quantification of alpha-1-antitrypsin by liquid chromatography-tandem mass spectrometry. AB - BACKGROUND: alpha-1-Antitrypsin (A1AT) deficiency results from a genetic disorder at 2 common loci. Diagnosis requires quantification of A1AT and subsequent identification of the specific variant. The current algorithm of laboratory testing for the diagnosis of A1AT deficiency uses a combination of quantification (nephelometry), genotyping, and/or phenotyping. We developed a multiple reaction monitoring liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous quantification of A1AT and identification of the 2 most common deficiency alleles present in 95% of the patients with A1AT deficiency. METHODS: Serum samples (n = 40) were digested with trypsin, and appropriate 13C/15N labeled standard peptides were added. We performed LC-MS/MS analysis with a 0.5- by 150-mm C18 column and H2O:acetonitrile:n-propanol:formic acid (A:98:1:1:0.2 and B:10:80:10:0.2; flow 12 MUL/min) mobile phase in positive ion mode on a TSQ Quantum triple quadrupole MS system. We measured the A1AT concentration by comparison to a calibration curve and determined the phenotype by the presence or absence of variant peptides. We compared the results to the current phenotyping assay by isoelectric focusing (IEF) and the immunonephelometry quantitative assay. RESULTS: For A1AT allele detection, in 39 of 40 samples the LC-MS/MS results were identical to those obtained by IEF gel electrophoresis. The single discrepant result was rerun by IEF at a lower dilution, and the results were in concordance. The A1AT quantification by LC-MS/MS also compared favorably with nephelometry. CONCLUSIONS: The LC-MS/MS method correlates well with current phenotyping and nephelometric assays and has the potential to improve the laboratory diagnosis of genetic A1AT deficiency. PMID- 21636699 TI - Molecular and cellular pharmacology of the novel noncamptothecin topoisomerase I inhibitor Genz-644282. AB - Camptothecin derivatives are powerful anticancer drugs because of their ability to trap topoisomerase I (Top1)-DNA cleavage complexes. However, they exhibit clinical limitations due to the instability of their alpha-hydroxylactone six membered E-ring structure. In addition, they exhibit bone marrow and intestinal toxicity, especially in adults, and are drug efflux substrates. Here, we report a novel Top1 inhibitor, Genz-644282. We show that Genz-644282 and its metabolites induce Top1 cleavage at similar, as well as unique genomic positions, compared with camptothecin. The compound also induces protein-linked DNA breaks and Top1 DNA cleavage complexes that persist longer after compound removal than camptothecin. Concentration-dependent and persistent gammaH2AX formation was readily observed in cells treated with Genz-644282, and was present in greater than 50% of the cell population following 24 hours compound exposure. The compound shows partial cross-resistance in cell lines resistant to camptothecin. These cell lines include the human prostate DU145RC0.1 and the leukemic CEM/C2 cells. Limited cross-resistance to Genz-644282 was also found in the Top1 knockdown colon cancer (HCT116) and breast cancer (MCF7) cell lines and in human adenocarcinoma cells (KB31/KBV1) that overexpress (P-glycoprotein, ABCB1), a member of the ATP-binding cassette family of cell surface transport proteins known to confer MDR. Together, our results provide the first molecular and cellular characterization of Genz-644282 and its clinically relevant metabolites. PMID- 21636700 TI - Leptin deficiency suppresses MMTV-Wnt-1 mammary tumor growth in obese mice and abrogates tumor initiating cell survival. AB - Obesity increases both the risk and mortality associated with many types of cancer including that of the breast. In mice, obesity increases both incidence of spontaneous tumors and burden of transplanted tumors. Our findings identify leptin, an adipose secreted cytokine, in promoting increased mammary tumor burden in obese mice and provide a link between this adipokine and cancer. Using a transplantable tumor that develops spontaneously in the murine mammary tumor virus-Wnt-1 transgenic mice, we show that tumors transplanted into obese leptin receptor (LepRb)-deficient (db/db) mice grow to eight times the volume of tumors transplanted into lean wild-type (WT) mice. However, tumor outgrowth and overall tumor burden is reduced in obese, leptin-deficient (ob/ob) mice. The residual tumors in ob/ob mice contain fewer undifferentiated tumor cells (keratin 6 immunopositive) compared with WT or db/db mice. Furthermore, tumors in ob/ob mice contain fewer cells expressing phosphorylated Akt, a growth promoting kinase activated by the LepRb, compared with WT and db/db mice. In vivo limiting dilution analysis of residual tumors from ob/ob mice indicated reduced tumor initiating activity suggesting fewer cancer stem cells (CSCs). The tumor cell populations reduced by leptin deficiency were identified by fluorescence activated cell sorting and found to express LepRb. Finally, LepRb expressing tumor cells exhibit stem cell characteristics based on the ability to form tumorspheres in vitro and leptin promotes their survival. These studies provide critical new insight on the role of leptin in tumor growth and implicate LepRb as a CSC target. PMID- 21636701 TI - Different gene expression profiles in metastasizing midgut carcinoid tumors. AB - The genetic events leading the progression of midgut carcinoid tumors are largely unknown. The disease course varies from patient to patient, and there is a lack of reliable prognostic markers. In order to identify genes involved in tumor progression, gene expression profiling was performed on tumor specimens. Samples comprised 18 primary tumors, 17 lymph node (LN) metastases, and seven liver metastases from a total of 19 patients. Patients were grouped according to clinical data and histopathology into indolent or progressive course. RNA was subjected to a spotted oligo microarray and B-statistics were performed. Differentially expressed genes were verified using quantitative real-time PCR. Self-organizing maps demonstrated three clusters: 11 primary tumors separated in one cluster, five LN metastases in another cluster, whereas all seven liver metastases, seven primary, and 12 LN metastases formed a third cluster. There was no correlation between indolent and progressive behavior. The primary tumors with Ki67 >5%, with low frequency of the carcinoid syndrome, and a tendency toward shorter survival grouped together. Primary tumors differed in expression profile from their associated LN metastases; thus, there is evidence for genetic changes from primary tumors to metastases. ACTG2, GREM2, REG3A, TUSC2, RUNX1, TPH1, TGFBR2, and CDH6 were differentially expressed between clusters and subgroups of tumors. The expression profile that assembles tumors as being genetically similar on the RNA expression level may not be concordant with the clinical disease course. This study reveals differences in gene expression profiles and novel genes that may be of importance in midgut carcinoid tumor progression. PMID- 21636702 TI - Novel candidate colorectal cancer biomarkers identified by methylation microarray based scanning. AB - DNA hypermethylation is a common epigenetic abnormality in colorectal cancers (CRCs) and a promising class of CRC screening biomarkers. We conducted a genome wide search for novel neoplasia-specific hypermethylation events in the colon. We applied methylation microarray analysis to identify loci hypermethylated in 17 primary CRCs relative to eight non-neoplastic colonic mucosae (NCs) from neoplasia-free subjects. These CRC-associated hypermethylation events were then individually evaluated for their ability to discriminate neoplastic from non neoplastic cases, based on real-time quantitative methylation-specific PCR (qMSP) assays in 113 colonic tissues: 51 CRCs, nine adenomas, 19 NCs from CRC patients (CRC-NCs), and 34 NCs from neoplasia-free subjects (control NCs). A strict microarray data filtering identified 169 candidate CRC-associated hypermethylation events. Fourteen of these 169 loci were evaluated using qMSP assays. Ten of these 14 methylation events significantly distinguished CRCs from age-matched control NCs (P<0.05 by receiver operator characteristic curve analysis); methylation of visual system homeobox 2 (VSX2) achieved the highest discriminative accuracy (83.3% sensitivity and 92.3% specificity, P<1*10(-6)), followed by BEN domain containing 4 (BEND4), neuronal pentraxin I (NPTX1), ALX homeobox 3 (ALX3), miR-34b, glucagon-like peptide 1 receptor (GLP1R), BTG4, homer homolog 2 (HOMER2), zinc finger protein 583 (ZNF583), and gap junction protein, gamma 1 (GJC1). Adenomas were significantly discriminated from control NCs by hypermethylation of VSX2, BEND4, NPTX1, miR-34b, GLP1R, and HOMER2 (P<0.05). CRC NCs were significantly distinguished from control NCs by methylation of ALX3 (P<1*10(-4)). In conclusion, systematic methylome-wide analysis has identified ten novel methylation events in neoplastic and non-neoplastic colonic mucosae from CRC patients. These potential biomarkers significantly discriminate CRC patients from controls. Thus, they merit further evaluation in stool- and circulating DNA-based CRC detection studies. PMID- 21636703 TI - Relationship between self-reported racial composition of high school and health literacy among community health center patients. AB - Intervention and policy approaches targeting the societal factors that affect health literacy (e.g., educational systems) could have promise to improve health outcomes, but little research has investigated these factors. This study examined the associations between self-reported racial composition of prior educational and neighborhood contexts and health literacy among 1,061 English- and Spanish speaking adult community health center patients. The authors found that self reported racial composition of high school was a significant predictor of health literacy among those who received schooling in the United States, controlling for race/ethnicity, education, age, country of birth, and survey language. Black and Hispanic patients had significantly lower health literacy than White patients within educational strata among those schooled in the United States. The findings revealed substantial disparities in health literacy. Self-reported racial composition of school context was a significant predictor of health literacy. Transdisciplinary, multilevel intervention approaches are likely to be needed to address the health literacy needs of this population. PMID- 21636704 TI - Neurocognitive function: an emerging surrogate endpoint for neuro-oncology trials. PMID- 21636705 TI - Pathway inhibition: emerging molecular targets for treating glioblastoma. AB - Insights into the molecular pathogenesis of glioblastoma have not yet resulted in relevant clinical improvement. With standard therapy, which consists of surgical resection with concomitant temozolomide in addition to radiotherapy followed by adjuvant temozolomide, the median duration of survival is 12-14 months. Therefore, the identification of novel molecular targets and inhibitory agents has become a focus of research for glioblastoma treatment. Recent results of bevacizumab may represent a proof of principle that treatment with targeted agents can result in clinical benefits for patients with glioblastoma. This review discusses limitations in the existing therapy for glioblastoma and provides an overview of current efforts to identify molecular targets using large scale screening of glioblastoma cell lines and tumor samples. We discuss preclinical and clinical data for several novel molecular targets, including growth factor receptors, phosphatidylinositol-3 kinase, SRC-family kinases, integrins, and CD95 ligand and agents that inhibit these targets, including erlotinib, enzastaurin, dasatinib, sorafenib, cilengitide, AMG102, and APG101. By combining advances in tumor screening with novel targeted therapies, it is hoped that new treatment options will emerge for this challenging tumor type. PMID- 21636706 TI - Downregulation of Pdcd4 by mir-21 facilitates glioblastoma proliferation in vivo. AB - MicroRNAs (miRNAs) are small, noncoding RNAs that play a critical role in developmental and physiological processes and are implicated in the pathogenesis of several human diseases, including cancer. They function by regulating target gene expression post-transcriptionally. In this study, we examined the role of oncogenic mir-21 in the pathogenesis of glioblastoma, the most aggressive form of primary brain tumor. We have previously reported that mir-21 is expressed at higher levels in primary glioblastoma-tissue and glioblastoma-derived cell lines than in normal brain tissue. We demonstrate that downregulation of mir-21 in glioblastoma-derived cell lines results in increased expression of its target, programmed cell death 4 (Pdcd4), a known tumor-suppressor gene. In addition, our data indicate that either downregulation of mir-21 or overexpression of its target, Pdcd4, in glioblastoma-derived cell lines leads to decreased proliferation, increased apoptosis, and decreased colony formation in soft agar. Using a glioblastoma xenograft model in immune-deficient nude mice, we observe that glioblastoma-derived cell lines in which mir-21 levels are downregulated or Pdcd4 is over-expressed exhibit decreased tumor formation and growth. Significantly, tumors grow when the glioblastoma-derived cell lines are transfected with anti-mir-21 and siRNA to Pdcd4, confirming that the tumor growth is specifically regulated by Pdcd4. These critical in vivo findings demonstrate an important functional linkage between mir-21 and Pdcd4 and further elucidate the molecular mechanisms by which the known high level of mir-21 expression in glioblastoma can attribute to tumorigenesis--namely, inhibition of Pdcd4 and its tumor-suppressive functions. PMID- 21636707 TI - Myeloid-derived suppressor cell accumulation and function in patients with newly diagnosed glioblastoma. AB - To assess the accumulation of myeloid-derived suppressor cells (MDSCs) in the peripheral blood of patients with glioma and to define their heterogeneity and their immunosuppressive function. Peripheral blood mononuclear cells (PBMCs) from healthy control subjects and from patients with newly diagnosed glioma were stimulated with anti-CD3/anti-CD28 and T cells assessed for intracellular expression of interferon (IFN)-gamma. Antibody staining of PBMCs from glioma patients and healthy donors (CD33, HLADR, CD15, and CD14) followed by 4-color flow cytometry analysis-defined MDSC levels in the peripheral blood. To assess the role of MDSCs in suppressing T cell IFNgamma production, PBMCs were depleted of MDSCs using anti-CD33 and anti-CD15 antibody-coated beads prior to T cell stimulation. Enzyme-linked immunosorbent assays were used to assess plasma arginase activity and the level of granulocyte colony-stimulating factor (G-CSF). Patients with glioblastoma have increased MDSC counts (CD33+HLADR-) in their blood that are composed of neutrophilic (CD15(+); >60%), lineage-negative (CD15( )CD14(-); 31%), and monocytic (CD14(+); 6%) subsets. After stimulation, T cells from patients with glioblastoma had suppressed IFN-gamma production when compared with healthy, age-matched donor T cells. Removal of MDSCs from the PBMCs with anti-CD33/CD15-coated beads significantly restored T cell function. Significant increases in arginase activity and G-CSF levels were observed in plasma specimens obtained from patients with glioblastoma. The accumulation of MDSCs in peripheral blood in patients with glioma likely promotes T cell immune suppression that is observed in this patient population. Increased plasma levels of arginase and G CSF may relate to MDSC suppressor function and MDSC expansion, respectively, in patients with glioma. PMID- 21636708 TI - High beta-catenin/Tcf-4 activity confers glioma progression via direct regulation of AKT2 gene expression. AB - Recent data suggest that the beta-catenin/Tcf-4 signaling pathway plays an important role in human cancer tumorigenesis. However, the mechanism of beta catenin/Tcf-4 signaling in tumorigenesis is poorly understood. In this study, we show that Tcf-4 protein levels were significantly elevated in high-grade gliomas in comparison with low-grade gliomas and that Tcf-4 levels correlated with levels of AKT2. Reduction of beta-catenin/Tcf-4 activity inhibited glioma cell proliferation and invasion in vitro and tumor growth in vivo. This effect of beta catenin/Tcf-4 activity was mediated by AKT2, and in vivo binding of beta catenin/Tcf-4 to the AKT2 promoter was validated using the chromatin immunoprecipitation assay and luciferase reporter assays. Taken together, we have demonstrated that Tcf-4 is associated with glioma progression and that AKT2 is a new member of the genes that are regulated by beta-catenin/Tcf-4. PMID- 21636709 TI - Control of proliferation in astrocytoma cells by the receptor tyrosine kinase/PI3K/AKT signaling axis and the use of PI-103 and TCN as potential anti astrocytoma therapies. AB - A growing body of work suggests that astrocytomas and glioblastoma multiforme will require carefully tailored, molecularly targeted therapy for successful treatment. Recent efforts to comprehensively identify mutations and gene expression changes in glioblastoma have shown that mutation of NF1 is a common alteration in human glioblastoma. We have developed and characterized a panel of 14 tumor lines from grades II through IV astrocytomas developed from our Nf1 /+;Trp53-/+cis mouse model and have used this panel to characterize signal transduction pathways and inhibitors that are candidate therapeutic targets for astrocytoma and glioblastoma. We show that these tumors express platelet-derived growth factor receptor-alpha, epidermal growth factor receptor, and their respective ligands to varying degrees. We find that both the MEK and PI3K signaling pathways downstream of epidermal growth factor receptor and platelet derived growth factor receptor-alpha are necessary for full proliferation of astrocytoma cells; however, inhibition of the PI3K pathway is more effective than inhibition of MEK at blocking cell growth. We have examined inhibitors of the PI3K/Akt/mTOR signaling pathway and find that PI-103 and TCN show particular promise for inhibiting growth in Nf1 and Trp53 mutant astrocytoma cells. PMID- 21636710 TI - International retrospective study of over 1000 adults with anaplastic oligodendroglial tumors. AB - Treatment for newly diagnosed anaplastic oligodendroglial tumors is controversial. Radiotherapy (RT) alone and in combination with chemotherapy (CT) are the most well studied strategies. However, CT alone is often advocated, especially in cases with 1p19q codeletion. We retrospectively identified 1013 adults diagnosed from 1981-2007 treated initially with RT alone (n = 200), CT + RT (n = 528), CT alone (n = 201), or other strategies (n = 84). Median overall survival (OS) was 6.3 years and time to progression (TTP) was 3.1 years. 1p19q codeletion correlated with longer OS and TTP than no 1p or 19q deletion. In codeleted cases, median TTP was longer following CT + RT (7.2 y) than following CT (3.9 y, P = .003) or RT (2.5 y, P < .001) alone but without improved OS; median TTP was longer following treatment with PCV alone than temozolomide alone (7.6 vs. 3.3 y, P = .019). In cases with no deletion, median TTP was longer following CT + RT (3.1 y) than CT (0.9 y, P = .0124) or RT (1.1 y, P < .0001) alone; OS also favored CT + RT (median 5.0 y) over CT (2.2 y, P = .02) or RT (1.9 y, P < .0001) alone. In codeleted cases, CT alone did not appear to shorten OS in comparison with CT + RT, and PCV appeared to offer longer disease control than temozolomide but without a clear survival advantage. Combined CT + RT led to longer disease control and survival than did CT or RT alone in cases with no 1p19q deletion. Ongoing trials will address these issues prospectively. PMID- 21636711 TI - Treatment of young children with localized medulloblastoma by chemotherapy alone: results of the prospective, multicenter trial HIT 2000 confirming the prognostic impact of histology. AB - This study was designed to confirm the previously observed favorable survival rates and prognostic factors in young children with nonmetastatic medulloblastoma (MB) treated with postoperative chemotherapy alone. Patients who received a diagnosis during the period January 2001 through December 2005 and who were aged <4 years received 3 cycles of postoperative systemic multiagent chemotherapy and intraventricular methotrexate. In cases of complete remission, treatment was terminated after 2 additional cycles of chemotherapy. Otherwise, secondary surgery, radiotherapy, and consolidation chemotherapy were recommended. At a median follow-up of 4.5 years, the 5-year event-free survival (EFS) and overall survival (OS) rates (+/- standard error) for 45 patients (median age, 2.5 years) were 57% +/- 8% and 80% +/- 6%, respectively. Nineteen patients with desmoplastic/nodular MB variants had better 5-year EFS and OS rates (90% +/- 7% and 100% +/- 0%, respectively) than did 23 patients with classic MB (30% +/- 11% and 68% +/- 10%, respectively; P < .001 for EFS; P = .008 for OS). Five-year EFS and OS rates for 3 children with anaplastic MB were 33% +/- 27%. Desmoplastic/nodular histology was an independent prognostic factor for EFS. Twenty-nine of 30 patients without postoperative residual tumor remained in continuous complete remission. Our results confirm that histology of MB variants is a strong prognostic factor in this age group. Sustained tumor control can be achieved by this chemotherapy regimen in young children with desmoplastic/nodular MB variants. For children with non-desmoplastic/nonnodular MB variants, for which predominantly local relapses lead to less favorable survival rates, local radiotherapy has been introduced after chemotherapy since 2006. PMID- 21636712 TI - Thalamic high-grade gliomas in children: a distinct clinical subset? AB - Pediatric high-grade gliomas (HGGs) of the thalamic region account for up to 13% of pediatric HGGs and usually result in only anecdotal long-term survival. Because very little is known about these tumors, we aimed to further characterize them. In our series of 99 pediatric thalamic HGGs, there were no significant differences in survival between patients with tumors affecting the thalamus alone (including bithalamic lesions) and patients with tumors affecting the thalamus plus adjacent structures. Tumor resection (event-free survival/overall survival) and an early treatment response to radiotherapy/chemotherapy (event-free survival) had independent prognostic significance, as shown by Kaplan-Meier and multivariate Cox regression analyses. When we compared clinical characteristics and outcomes of pediatric thalamic HGG with those of pediatric (nonthalamic) supratentorial (n = 177) as well as pediatric pontine HGG (including diffuse intrinsic pontine gliomas; n = 234), we found that thalamic HGG shared more similarities with pontine than with supratentorial HGG, but overall, it appeared to represent a clinically distinct subgroup of pediatric HGG. The varying extent of tumor resection in the different tumor localizations may play some role in the observed clinical differences, as shown by multivariate Cox regression analyses, but the tumor site itself was also identified as an independent prognostic parameter. Thus, an additional location-specific effect on survival and/or tumor biology, despite different neurosurgical accessibility, has to be considered. Therefore, future investigations should try to further characterize the obviously site-specific heterogeneity of pediatric HGG on a molecular genetic basis. PMID- 21636713 TI - Sema3A [corrected] resists retinal revascularization. AB - In this issue of Blood, Joyal and colleagues make the insightful finding that Semaphorin3A (Sema3A) is secreted by hypoxic neurons in ischemic/avascular retina,thereby inhibiting vascular regeneration of the retina and enhancing pathologic preretinal neovascularization. PMID- 21636714 TI - For disease in need, a Friend indeed. AB - It has been 40 years since Charlotte Friend demonstrated the unbelievable for the second time. PMID- 21636715 TI - "Let"-ing go with clonal expansion? AB - In this issue of Blood, Ikeda et al report the effects of the high mobility group A2 (Deltahmga2) lacking the 3' untranslated region (UTR) in hematopoietic tissues. Overexpression of Deltahmga2 in transgenic mice resulted in "big mice with big blood": the mice weighed more and had a myeloproliferative phenotype with increases in peripheral blood counts, splenomegaly, a hypercellular bone marrow and erythropoietin-independent erythroid colony formation. PMID- 21636716 TI - When differentiation goes viral. AB - In this issue of Blood, Vrzalikova et al present evidence that Epstein-Barr virus (EBV) infection leads to the down-regulation of BLIMP1 in primary germinal center (GC) B cells. BLIMP1 is a transcription factor encoded by the PRDM1 gene. It stops cell cycling and leads to terminal differentiation of B cells to plasma cells. The authors suggest that EBV inhibition of BLIMP1 expression serves to inhibit terminal differentiation of EBV-infected cells. PMID- 21636717 TI - Megakaryocyte TLR2: immunity bullet? AB - Innate immunity is the host defensive weapon against pathogens and this issue of Blood offers an interesting new bullet for the immune system gun by studying the role of Toll-like receptor 2 (TLR2) in megakaryocytic function; a field previously not explored. Beaulieu and colleagues provide evidence that inflammation, through TLR2, can increase megakaryocyte maturation and modulate its phenotype, contributing to our understanding of the interrelationship between inflammation and hemostasis. PMID- 21636718 TI - A novel mechanism of transplacental cancer transmission: natural killer/T-cell lymphoma in the paratesticular region is of maternal origin. PMID- 21636719 TI - Fanconi anemia (FA)-associated 3q gains in leukemic transformation consistently target EVI1, but do not affect low TERC expression in FA. PMID- 21636720 TI - Doubts concerning the recently reported human neutrophil lifespan of 5.4 days. PMID- 21636721 TI - Deuterium and neutrophil kinetics. PMID- 21636722 TI - Anemia in congenital nephrotic syndrome: role of urinary copper and ceruloplasmin loss. PMID- 21636724 TI - Rotation constraint after double-bundle ACL reconstruction: letter/response. PMID- 21636723 TI - Extracellular nucleotides and apyrases regulate stomatal aperture in Arabidopsis. AB - This study investigates the role of extracellular nucleotides and apyrase enzymes in regulating stomatal aperture. Prior data indicate that the expression of two apyrases in Arabidopsis (Arabidopsis thaliana), APY1 and APY2, is strongly correlated with cell growth and secretory activity. Both are expressed strongly in guard cell protoplasts, as determined by reverse transcription-polymerase chain reaction and immunoblot analyses. Promoter activity assays for APY1 and APY2 show that expression of both apyrases correlates with conditions that favor stomatal opening. Correspondingly, immunoblot data indicate that APY expression in guard cell protoplasts rises quickly when these cells are moved from darkness into light. Both short-term inhibition of ectoapyrase activity by polyclonal antibodies and long-term suppression of APY1 and APY2 transcript levels significantly disrupt normal stomatal behavior in light. Stomatal aperture shows a biphasic response to applied adenosine 5'-[gamma-thio]triphosphate (ATPgammaS) or adenosine 5'-[beta-thio] diphosphate, with lower concentrations inducing stomatal opening and higher concentrations inducing closure. Equivalent concentrations of adenosine 5'-O-thiomonophosphate have no effect on aperture. Two mammalian purinoceptor inhibitors block ATPgammaS- and adenosine 5'-[beta thio] diphosphate-induced opening and closing and also partially block the ability of abscisic acid to induce stomatal closure and of light to induce stomatal opening. Treatment of epidermal peels with ATPgammaS induces increased levels of nitric oxide and reactive oxygen species, and genetically suppressing the synthesis of these agents blocks the effects of nucleotides on stomatal aperture. A luciferase assay indicates that treatments that induce either the closing or opening of stomates also induce the release of ATP from guard cells. These data favor the novel conclusion that ectoapyrases and extracellular nucleotides play key roles in regulating stomatal functions. PMID- 21636725 TI - Predifferentiated mesenchymal stem cells for osteochondral defects: letter. PMID- 21636726 TI - Medial patellofemoral attachment site in children: letter/response. PMID- 21636727 TI - Medial patellofemoral ligament injury in children: letter. PMID- 21636728 TI - Platelet-rich plasma augmentation of rotator cuff repair: letter. PMID- 21636729 TI - Keeping it simple. PMID- 21636730 TI - Achievement motivation and memory: achievement goals differentially influence immediate and delayed remember-know recognition memory. AB - Little research has been conducted on achievement motivation and memory and, more specifically, on achievement goals and memory. In the present research, the authors conducted two experiments designed to examine the influence of mastery approach and performance-approach goals on immediate and delayed remember-know recognition memory. The experiments revealed differential effects for achievement goals over time: Performance-approach goals showed higher correct remember responding on an immediate recognition test, whereas mastery-approach goals showed higher correct remember responding on a delayed recognition test. Achievement goals had no influence on overall recognition memory and no consistent influence on know responding across experiments. These findings indicate that it is important to consider quality, not just quantity, in both motivation and memory, when studying relations between these constructs. PMID- 21636731 TI - Does being attractive always help? Positive and negative effects of attractiveness on social decision making. AB - Previous studies of organizational decision making demonstrate an abundance of positive biases directed toward highly attractive individuals. The current research, in contrast, suggests that when the person being evaluated is of the same sex as the evaluator, attractiveness hurts, rather than helps. Three experiments assessing evaluations of potential job candidates (Studies 1 and 3) and university applicants (Study 2) demonstrated positive biases toward highly attractive other-sex targets but negative biases toward highly attractive same sex targets. This pattern was mediated by variability in participants' desire to interact with versus avoid the target individual (Studies 1 and 2) and was moderated by participants' level of self-esteem (Study 3); the derogation of attractive same-sex targets was not observed among people with high self-esteem. Findings demonstrate an important exception to the positive effects of attractiveness in organizational settings and suggest that negative responses to attractive same-sex targets stem from perceptions of self-threat. PMID- 21636732 TI - Effects of varicocelectomy on sperm DNA fragmentation, mitochondrial function, chromatin condensation, and apoptosis. AB - The aim of this study was to evaluate conventional semen parameters (density, morphology, and progressive motility) and the flow-cytometric parameters of DNA fragmentation, mitochondrial membrane potential, phosphatidylserine externalization, and chromatin compactness in patients with varicocele before and after varicocelectomy. Thirty men (26.5 +/- 3.2 years old, range 20-32 years) with oligoasthenoteratozoospermia and grade 3 left varicocele were selected (without other causes of male infertility). Each of them underwent sperm analysis and flow cytometric evaluation before and 4 months after subinguinal microsurgical varicocelectomy (SMV). After varicocelectomy, men had significantly higher sperm density, progressive motility, and normal forms compared with baseline. They also had a significantly lower percentage of spermatozoa with low mitochondrial membrane potential. After SMV, they showed a significantly lower percentage of spermatozoa with phosphatidylserine externalization, an early sign of apoptosis. Significantly decreased percentages of spermatozoa with abnormal chromatin compactness and spermatozoa with DNA fragmentation were found after SMV compared with baseline. Subinguinal microsurgical varicocelectomy improves sperm function in oligoasthenoteratozoospermia secondary to grade 3 left varicocele. Improvements are seen in conventional parameters and biofunctional parameters not routinely evaluated. PMID- 21636733 TI - Donor sperm insemination cycles: are two inseminations better than one? AB - The objective of this study was to determine the clinical pregnancy rate with 2 inseminations compared to a single intrauterine insemination (IUI) in a given cycle using frozen-thawed donor sperm. This was a retrospective study at a university practice; patients were women using donor sperm. We conducted a comparison of single IUI, intracervical insemination (ICI) followed by an IUI on the next day, and double IUI (2 consecutive days); clinical pregnancy rate was the main outcome measure. The cycle-specific and total pregnancy rates were not significantly different between the 3 protocol groups (306 cycles). The average pregnancy rate over 3 cycles was 10.2% for IUI, 15.3% for ICI/IUI, and 13.7% for IUI/IUI (P = .47). After controlling for repeated measures per subject and age, gravidity, and use of Clomid, there was no significant difference between protocols. The ICI/IUI (odds ratio [OR] = 1.70; 95% confidence interval [CI], 0.83-3.51) and IUI/IUI (OR = 1.5; 95% CI, 0.52-4.33) protocols appeared more likely to result in a clinical pregnancy than the single IUI protocol. Current information on the optimal number of inseminations per cycle using donor sperm is limited. Our large study using 3 protocols found an increase in pregnancy rate with the addition of either an ICI or IUI to a single IUI protocol in a natural or Clomid cycle but did not meet statistical significance. Additional prospective studies are needed to better counsel patients using donor sperm. PMID- 21636734 TI - Multiple penile schwannomas and their magnetic resonance imaging characteristics. AB - Penile schwannoma is a rare encapsulated nerve sheath tumor of the peripheral nerve in the penis. Here, we report a case of multiple, painful, beadlike, nodular tumors in the penis of a 40-year-old man. The nodules showed well defined margins and hypoechoic areas on gray-scale ultrasonograms. They showed high signal intensity on fat-saturated T2-weighted magnetic resonance imaging (MRI) and intense enhancement on gadolinium-enhanced T1-weighted MRI, compatible with their myxoid content. The nodules pressed against the tunica albuginea and some caused inward indentation, which we considered as indirect evidence of the tumor location. In conclusion, the MRI characteristics and location of the nodules together enable presurgical diagnosis of penile schwannomas, which is especially useful in asymptomatic patients and those unwilling to undergo surgery. PMID- 21636735 TI - The effects of honey supplementation on seminal plasma cytokines, oxidative stress biomarkers, and antioxidants during 8 weeks of intensive cycling training. AB - The purpose of this study was to examine the effects of natural honey supplementation on seminal plasma cytokines, oxidative stress biomarkers, and antioxidants during 8 weeks of intensive cycling training in male road cyclists. Thirty-nine healthy nonprofessional male road cyclists aged 18-28 years participated in this study. The participants were randomly assigned to exercise + supplement (E + S, n = 20) and exercise (E, n = 19) groups. All subjects participated in 8 weeks of intensive cycling training. Ninety minutes before each training session, subjects in the E + S group supplemented with 70 g of honey, whereas subjects in the E group received 70 g of an artificial sweetener. All subjects had an initial semen sampling at baseline (T(1)). The next 6 semen collections were collected immediately (T(2)) and 12 (T(3)) and 24 hours (T(4)) after the last training session in week 4, as well as immediately (T(5)) and 12 (T(6)) and 24 hours (T(7)) after the last training session in week 8, respectively. In the E group, 8 weeks of intensive cycling training significantly increased seminal interleukin (IL)-1beta, IL-6, IL-8, tumor necrosis factor (TNF)-alpha, reactive oxygen species (ROS), and malondialdehyde (MDA) levels (P < .008) and significantly decreased the levels of seminal superoxide dismutase (SOD), catalase, and total antioxidant capacity (TAC) (P < .008). Significantly less elevation in seminal IL-1beta, IL-6, IL-8, TNF-alpha, ROS, and MDA levels (P < .008) and significant increases in seminal SOD, catalase, and TAC concentrations were observed after the honey supplementation in the E + S group (P < .008). It may be possible that honey supplementation following long-term intensive cycling training would be effective in attenuating the probable aggravating effects of intensive cycling training on spermatogenesis and fertility capacity in road cyclists. PMID- 21636736 TI - Restricted expression of Ovol2/MOVO in XY body of mouse spermatocytes at the pachytene stage. AB - The development of multicellular organisms is controlled by sequential activation of a hierarchy of regulatory genes, which encode transcription factors having DNA-binding motifs. We previously identified a testis-specific zinc finger transcriptional factor, Ovol2/MOVO, as a mouse homologue of Drosophila Ovo. Because mice deficient in Ovol2/Movo die during early embryogenesis, its function in male germ cells has remained unknown. We have recently succeeded in preparing anti-Ovol2/MOVO antiserum for immunohistochemical use. In the present study, we demonstrated that Ovol2/MOVO protein started to be expressed in male germ cells at 2 weeks after birth and that Ovol2/MOVO expression was restricted to the XY body in spermatocytes at the pachytene stage. In a reporter assay, Ovol2/MOVO repressed the histone H1t promoter activity in the spermatogenic cell line GC-2spd. These results suggest that Ovol2/MOVO may play an important role in the XY body during spermatogenesis, possibly in the processes of XY body formation and meiotic sex chromosome inactivation. PMID- 21636737 TI - Single-nucleotide polymorphisms in the SEPTIN12 gene may be a genetic risk factor for Japanese patients with Sertoli cell-only syndrome. AB - Genetic mechanisms have been implicated as a cause of some cases of male infertility. Recently, 10 novel genes involved in human spermatogenesis, including human SEPTIN12, were identified by expression microarray analysis of human testicular tissue. Septin12 is a member of the septin family of conserved cytoskeletal GTPases that form heteropolymeric filamentous structures in interphase cells. It is expressed specifically in the testis. Therefore, we hypothesized that mutation or polymorphisms of SEPTIN12 participate in male infertility, especially Sertoli cell-only syndrome (SCOS). To investigate whether SEPTIN12 gene defects are associated with azoospermia caused by SCOS, mutational analysis was performed in 100 Japanese patients by direct sequencing of coding regions. Statistical analysis was performed in patients with SCOS and in 140 healthy control men. No mutations were found in SEPTIN12 ; however, 8 coding single-nucleotide polymorphisms (SNP1-SNP8) could be detected in the patients with SCOS. The genotype and allele frequencies in SNP3, SNP4, and SNP6 were notably higher in the SCOS group than in the control group (P < .001). These results suggest that SEPTIN12 might play a critical role in human spermatogenesis. PMID- 21636738 TI - Pituitary adenylate cyclase-activating polypeptide (PACAP) and its receptors in the zebrafish ovary: evidence for potentially dual roles of PACAP in controlling final oocyte maturation. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuropeptide originally purified from ovine hypothalamus for its potent activity to stimulate cAMP production. However, its presence and action have also been demonstrated in various peripheral tissues including the ovary. In the zebrafish, two forms of PACAP (PACAP(38)-1, adcyap1a; and PACAP(38)-2, adcyap1b) and three PACAP receptors (PAC(1)-R, adcyap1r1; VPAC(1)-R, vipr1; and VPAC(2)-R, vipr2) were all expressed in the ovary. Interestingly, although both follicle cells and oocytes express adcyap1b, the expression of adcyap1a was restricted to the oocytes only. Among the three receptors, adcyap1r1 and vipr2 were expressed in the oocytes, whereas the expression of vipr1 was exclusively located in the follicle cells. Temporal expression analysis of PACAP ligands and receptors during folliculogenesis suggested that PACAP might play differential roles in regulating follicle growth and maturation through different receptors. The two receptors that are expressed in the oocyte (adcyap1r1 and vipr2) showed a significant increase in expression at the transition from the primary growth (PG) stage to previtellogenic (PV) stage and their levels maintained high during follicle growth. However, when the follicle development approached full-grown (FG) stage, these two receptors both decreased significantly in expression. In contrast, vipr1, the receptor expressed in the follicle cells, showed little change in expression at the PG-PV transition and afterwards during follicle growth; however, its expression surged dramatically at the FG stage prior to oocyte maturation. Based on these results, we hypothesized that PACAP might play dual roles in regulating follicle growth and maturation through different receptors located in different compartments. PACAP may stimulate oocyte growth but block its maturation in early follicles by acting directly on the oocyte via PAC1-R and VPAC2-R, whose expression is dominant in growth phase; however, PACAP may promote oocyte maturation in the maturation phase via VPAC1-R on the follicle cells, whose expression surges in FG follicles prior to maturation and is consistently high in the follicles undergoing final maturation. This hypothesis was further supported by the observation that PACAP promoted maturation of follicle-enclosed oocytes but suppressed spontaneous maturation of denuded oocytes in vitro. This study provides strong evidence for a PACAP-mediated signaling network in the zebrafish ovarian follicle, which may play roles in orchestrating follicle growth and maturation via different types of receptors located in different compartments of the follicle. PMID- 21636739 TI - Bisphenol A increases mammary cancer risk in two distinct mouse models of breast cancer. AB - Bisphenol A (BPA) is an industrial plasticizer that leaches from food containers during normal usage, leading to human exposure. Early and chronic exposure to endocrine-disrupting environmental contaminants such as BPA elevates the potential for long-term health consequences. We examined the impact of BPA exposure on fetal programming of mammary tumor susceptibility as well as its growth promoting effects on transformed breast cancer cells in vivo. Fetal mice were exposed to 0, 25, or 250 MUg/kg BPA by oral gavage of pregnant dams. Offspring were subsequently treated with the known mammary carcinogen, 7,12 dimethylbenz[a]anthracene (DMBA). While no significant differences in postnatal mammary development were observed, both low- and high-dose BPA cohorts had a statistically significant increase in susceptibility to DMBA-induced tumors compared to vehicle-treated controls. To determine if BPA also promotes established tumor growth, MCF-7 human breast cancer cells were subcutaneously injected into flanks of ovariectomized NCR nu/nu female mice treated with BPA, 17beta-estradiol, or placebo alone or combined with tamoxifen. Both estradiol- and BPA-treated cohorts formed tumors by 7 wk post-transplantation, while no tumors were detected in the placebo cohort. Tamoxifen reversed the effects of estradiol and BPA. We conclude that BPA may increase mammary tumorigenesis through at least two mechanisms: molecular alteration of fetal glands without associated morphological changes and direct promotion of estrogen-dependent tumor cell growth. Both results indicate that exposure to BPA during various biological states increases the risk of developing mammary cancer in mice. PMID- 21636740 TI - Simulation of quality-adjusted survival in chronic diseases: an application in type 2 diabetes. AB - BACKGROUND: Recent studies have demonstrated that measures of health-related quality of life can predict complications and mortality in patients with diabetes, even after adjustment for clinical risk factors. METHODS: The authors developed a simulation model of disease progression in type 2 diabetes to investigate the impact of patient quality of life on lifetime outcomes and its potential response to therapy. Changes in health utility over time are captured as a result of complications and aging. All risk equations, model parameter estimates, and input data were derived from patient-level data from the Fenofibrate Intervention and Event Lowering in Diabetes (FIELD) trial. RESULTS: Healthier patients with type 2 diabetes enjoy more life years, quality-adjusted life years (QALYs), and more life years free of complications. A 65-year-old patient at full health (utility = 1) can expect to live approximately 2 years longer and achieve 6 more QALYs than a patient at average health (utility = 0.8), given similar clinical risk factors. For patients with higher EQ-5D utility, the additional years lived without complications contribute more to longer life expectancy than years lived with complications. CONCLUSIONS: The authors have developed a model for progression of disease in diabetes that has a number of novel features; it captures the observed relationships between measures of quality of life and future outcomes, the number of states have been minimized, and it can be parameterized with just 4 risk equations. Underlying the simple model structure is important patient-level heterogeneity in health and outcomes. The simulations suggest that differences in patients' EQ-5D utility can account for large differences in QALYs, which could be relevant in cost-utility analyses. PMID- 21636741 TI - The quenching effect of flavonoids on 4-methylumbelliferone, a potential pitfall in fluorimetric neuraminidase inhibition assays. AB - Many assays aimed to test the inhibitory effects of synthetic molecules, and naturally occurring products on the neuraminidase activity exploit the hydrolysis of 2'-O-(4-methylumbelliferyl)-N-acetylneuraminic acid (4-MUNANA). The amount of the released product, 4-methylumbelliferone (4-MU), is then measured fluorimetrically. The authors attempted an analysis of the inhibitory properties of 35 naturally occurring flavonoids on neuraminidase N3, where only 29 of them were sufficiently soluble in the assay medium. During the analysis, the authors noticed a strong quenching effect due to the test compounds on the fluorescence of 4-MU. The quenching constants for the flavonoids were determined according to the Stern-Volmer approach. The extent of fluorescence reduction due to quenching and the magnitude of the fluorescence reduction measured in the inhibition assays were comparable: for 11 of 29 compounds, the two values were found to be coincident within the experimental uncertainty. These data were statistically analyzed for correlation by calculating the pertinent Pearson correlation coefficient. Inhibition and quenching were found to be positively correlated (r = 0.71, p(uncorr) = 1.5 * 10(-5)), and the correlation was maintained for the whole set of tested compounds. Altogether, the collected data imply that all of the tested flavonoids could produce false-positive results in the neuraminidase inhibition assay using 4-MUNANA as a substrate. PMID- 21636742 TI - Detection of convective downflows in a sunspot penumbra. AB - The fine structure and dynamics of sunspots and the strong outflow in their outer filamentary part--the penumbra--have puzzled astronomers for more than a century. Recent theoretical models and three-dimensional numerical simulations explain the penumbral filaments and their radiative energy output as the result of overturning convection. Here, we describe the detection of ubiquitous, relatively dark downward flows of up to 1 kilometer per second (km/s) in the interior penumbra, using imaging spectropolarimetric data from the Swedish 1-meter Solar Telescope. The dark downflows are omnipresent in the interior penumbra, distinguishing them from flows in arched flux tubes, and are associated with strong (3 to 3.5 km/s) radial outflows. They are thus part of a penumbral convective flow pattern, with the Evershed flow representing the horizontal component of that convection. PMID- 21636743 TI - Adult neural function requires MeCP2. AB - Rett syndrome (RTT) is a postnatal neurological disorder caused by mutations in MECP2, encoding the epigenetic regulator methyl-CpG-binding protein 2 (MeCP2). The onset of RTT symptoms during early life together with findings suggesting neurodevelopmental abnormalities in RTT and mouse models of RTT raised the question of whether maintaining MeCP2 function exclusively during early life might protect against disease. We show by using an inducible model of RTT that deletion of Mecp2 in adult mice recapitulates the germline knock-out phenotype, underscoring the ongoing role of MeCP2 in adult neurological function. Moreover, unlike the effects of other epigenetic instructions programmed during early life, the effects of early MeCP2 function are lost soon after its deletion. These findings suggest that therapies for RTT must be maintained throughout life. PMID- 21636744 TI - Processive movement of MreB-associated cell wall biosynthetic complexes in bacteria. AB - The peptidoglycan cell wall and the actin-like MreB cytoskeleton are major determinants of cell shape in rod-shaped bacteria. The prevailing model postulates that helical, membrane-associated MreB filaments organize elongation specific peptidoglycan-synthesizing complexes along sidewalls. We used total internal reflection fluorescence microscopy to visualize the dynamic relation between MreB isoforms and cell wall synthesis in live Bacillus subtilis cells. During exponential growth, MreB proteins did not form helical structures. Instead, together with other morphogenetic factors, they assembled into discrete patches that moved processively along peripheral tracks perpendicular to the cell axis. Patch motility was largely powered by cell wall synthesis, and MreB polymers restricted diffusion of patch components in the membrane and oriented patch motion. PMID- 21636745 TI - Coupled, circumferential motions of the cell wall synthesis machinery and MreB filaments in B. subtilis. AB - Rod-shaped bacteria elongate by the action of cell wall synthesis complexes linked to underlying dynamic MreB filaments. To understand how the movements of these filaments relate to cell wall synthesis, we characterized the dynamics of MreB and the cell wall elongation machinery using high-precision particle tracking in Bacillus subtilis. We found that MreB and the elongation machinery moved circumferentially around the cell, perpendicular to its length, with nearby synthesis complexes and MreB filaments moving independently in both directions. Inhibition of cell wall synthesis by various methods blocked the movement of MreB. Thus, bacteria elongate by the uncoordinated, circumferential movements of synthetic complexes that insert radial hoops of new peptidoglycan during their transit, possibly driving the motion of the underlying MreB filaments. PMID- 21636746 TI - The values of science. PMID- 21636750 TI - Italy. Quake experts to be tried for manslaughter. PMID- 21636751 TI - Microbiology. Concerns about arsenic-laden bacterium aired. PMID- 21636752 TI - Biomedicine. Wellcome puts its money on elite researchers. PMID- 21636753 TI - Ecology. Appraising U.K. ecosystems, report envisions greener horizon. PMID- 21636754 TI - DNA nanotechnology. DNA nanotechnology grows up. PMID- 21636755 TI - DNA nanotechnology. Next step: DNA robots? PMID- 21636756 TI - Physics. Possible sighting of dark matter fires up search and tempers. PMID- 21636757 TI - Zoos and captive breeding. PMID- 21636758 TI - Editor's note. PMID- 21636760 TI - Agriculture. Research principles for developing country food value chains. PMID- 21636761 TI - Biochemistry. Scaling up DNA computation. PMID- 21636762 TI - Planetary science. Io's tortured interior. PMID- 21636763 TI - Materials science. Potential solutions for creating responsive materials. PMID- 21636764 TI - Evolution. In evolution, the sum is less than its parts. PMID- 21636765 TI - Genomics. Behavior and the dynamic genome. PMID- 21636766 TI - Polariton superfluids reveal quantum hydrodynamic solitons. AB - A quantum fluid passing an obstacle behaves differently from a classical one. When the flow is slow enough, the quantum gas enters a superfluid regime, and neither whirlpools nor waves form around the obstacle. For higher flow velocities, it has been predicted that the perturbation induced by the defect gives rise to the turbulent emission of quantized vortices and to the nucleation of solitons. Using an interacting Bose gas of exciton-polaritons in a semiconductor microcavity, we report the transition from superfluidity to the hydrodynamic formation of oblique dark solitons and vortex streets in the wake of a potential barrier. The direct observation of these topological excitations provides key information on the mechanisms of superflow and shows the potential of polariton condensates for quantum turbulence studies. PMID- 21636767 TI - Observing the average trajectories of single photons in a two-slit interferometer. AB - A consequence of the quantum mechanical uncertainty principle is that one may not discuss the path or "trajectory" that a quantum particle takes, because any measurement of position irrevocably disturbs the momentum, and vice versa. Using weak measurements, however, it is possible to operationally define a set of trajectories for an ensemble of quantum particles. We sent single photons emitted by a quantum dot through a double-slit interferometer and reconstructed these trajectories by performing a weak measurement of the photon momentum, postselected according to the result of a strong measurement of photon position in a series of planes. The results provide an observationally grounded description of the propagation of subensembles of quantum particles in a two-slit interferometer. PMID- 21636768 TI - Two-dimensional Mott-Hubbard electrons in an artificial honeycomb lattice. AB - Artificial crystal lattices can be used to tune repulsive Coulomb interactions between electrons. We trapped electrons, confined as a two-dimensional gas in a gallium arsenide quantum well, in a nanofabricated lattice with honeycomb geometry. We probed the excitation spectrum in a magnetic field, identifying collective modes that emerged from the Coulomb interaction in the artificial lattice, as predicted by the Mott-Hubbard model. These observations allow us to determine the Hubbard gap and suggest the existence of a Coulomb-driven ground state. PMID- 21636769 TI - A material with electrically tunable strength and flow stress. AB - The selection of a structural material requires a compromise between strength and ductility. The material properties will then be set by the choice of alloy composition and microstructure during synthesis and processing, although the requirements may change during service life. Materials design strategies that allow for a recoverable tuning of the mechanical properties would thus be desirable, either in response to external control signals or in the form of a spontaneous adaptation, for instance in self-healing. We have designed a material that has a hybrid nanostructure consisting of a strong metal backbone that is interpenetrated by an electrolyte as the second component. By polarizing the internal interface via an applied electric potential, we accomplish fast and repeatable tuning of yield strength, flow stress, and ductility. The concept allows the user to select, for instance, a soft and ductile state for processing and a high-strength state for service as a structural material. PMID- 21636770 TI - Magnetosphere sawtooth oscillations induced by ionospheric outflow. AB - The sawtooth mode of convection of Earth's magnetosphere is a 2- to 4-hour planetary-scale oscillation powered by the solar wind-magnetosphere-ionosphere (SW-M-I) interaction. Using global simulations of geospace, we have shown that ionospheric O(+) outflows can generate sawtooth oscillations. As the outflowing ions fill the inner magnetosphere, their pressure distends the nightside magnetic field. When the outflow fluence exceeds a threshold, magnetic field tension cannot confine the accumulating fluid; an O(+)-rich plasmoid is ejected, and the field dipolarizes. Below the threshold, the magnetosphere undergoes quasi-steady convection. Repetition and the sawtooth period are controlled by the strength of the SW-M-I interaction, which regulates the outflow fluence. PMID- 21636771 TI - Diminishing returns epistasis among beneficial mutations decelerates adaptation. AB - Epistasis has substantial impacts on evolution, in particular, the rate of adaptation. We generated combinations of beneficial mutations that arose in a lineage during rapid adaptation of a bacterium whose growth depended on a newly introduced metabolic pathway. The proportional selective benefit for three of the four loci consistently decreased when they were introduced onto more fit backgrounds. These three alleles all reduced morphological defects caused by expression of the foreign pathway. A simple theoretical model segregating the apparent contribution of individual alleles to benefits and costs effectively predicted the interactions between them. These results provide the first evidence that patterns of epistasis may differ for within- and between-gene interactions during adaptation and that diminishing returns epistasis contributes to the consistent observation of decelerating fitness gains during adaptation. PMID- 21636772 TI - Negative epistasis between beneficial mutations in an evolving bacterial population. AB - Epistatic interactions between mutations play a prominent role in evolutionary theories. Many studies have found that epistasis is widespread, but they have rarely considered beneficial mutations. We analyzed the effects of epistasis on fitness for the first five mutations to fix in an experimental population of Escherichia coli. Epistasis depended on the effects of the combined mutations- the larger the expected benefit, the more negative the epistatic effect. Epistasis thus tended to produce diminishing returns with genotype fitness, although interactions involving one particular mutation had the opposite effect. These data support models in which negative epistasis contributes to declining rates of adaptation over time. Sign epistasis was rare in this genome-wide study, in contrast to its prevalence in an earlier study of mutations in a single gene. PMID- 21636773 TI - Scaling up digital circuit computation with DNA strand displacement cascades. AB - To construct sophisticated biochemical circuits from scratch, one needs to understand how simple the building blocks can be and how robustly such circuits can scale up. Using a simple DNA reaction mechanism based on a reversible strand displacement process, we experimentally demonstrated several digital logic circuits, culminating in a four-bit square-root circuit that comprises 130 DNA strands. These multilayer circuits include thresholding and catalysis within every logical operation to perform digital signal restoration, which enables fast and reliable function in large circuits with roughly constant switching time and linear signal propagation delays. The design naturally incorporates other crucial elements for large-scale circuitry, such as general debugging tools, parallel circuit preparation, and an abstraction hierarchy supported by an automated circuit compiler. PMID- 21636774 TI - Residue-specific vibrational echoes yield 3D structures of a transmembrane helix dimer. AB - Two-dimensional (2D) vibrational echo spectroscopy has previously been applied to structural determination of small peptides. Here we extend the technique to a more complex, biologically important system: the homodimeric transmembrane dimer from the alpha chain of the integrin alpha(IIb)beta(3). We prepared micelle suspensions of the pair of 30-residue chains that span the membrane in the native structure, with varying levels of heavy ((13)C=(18)O) isotopes substituted in the backbone of the central 10th through 20th positions. The constraints derived from vibrational coupling of the precisely spaced heavy residues led to determination of an optimized structure from a range of model candidates: Glycine residues at the 12th, 15th, and 16th positions form a tertiary contact in parallel right handed helix dimers with crossing angles of -58 degrees +/- 9 degrees and interhelical distances of 7.7 +/- 0.5 angstroms. The frequency correlation established the dynamical model used in the analysis, and it indicated the absence of mobile water associated with labeled residues. Delocalization of vibrational excitations between the helices was also quantitatively established. PMID- 21636775 TI - Interaction between Notch and Hif-alpha in development and survival of Drosophila blood cells. AB - A blood cell type termed crystal cell in Drosophila functions in clotting and wound healing and requires Notch for specification and maintenance. We report that crystal cells express elevated levels of Sima protein orthologous to mammalian hypoxia-inducible factor-alpha (Hif-alpha) even under conditions of normal oxygen availability. In these platelet-like crystal cells, Sima activates full-length Notch receptor signaling via a noncanonical, ligand-independent mechanism that promotes hemocyte survival during both normal hematopoietic development and hypoxic stress. This interaction initiates in early endosomes, is independent of Hif-beta (Tauangomicron in Drosophila), and does not activate hypoxia response targets. Studies in vertebrate myeloid cells have shown a similar up-regulation of Hif-alpha protein in well-oxygenated environments. This study provides a mechanistic paradigm for Hif-alpha/Notch interaction that may be conserved in mammals. PMID- 21636776 TI - Increased structure and active learning reduce the achievement gap in introductory biology. AB - Science, technology, engineering, and mathematics instructors have been charged with improving the performance and retention of students from diverse backgrounds. To date, programs that close the achievement gap between students from disadvantaged versus nondisadvantaged educational backgrounds have required extensive extramural funding. We show that a highly structured course design, based on daily and weekly practice with problem-solving, data analysis, and other higher-order cognitive skills, improved the performance of all students in a college-level introductory biology class and reduced the achievement gap between disadvantaged and nondisadvantaged students--without increased expenditures. These results support the Carnegie Hall hypothesis: Intensive practice, via active-learning exercises, has a disproportionate benefit for capable but poorly prepared students. PMID- 21636777 TI - Pediatric restless legs syndrome: analysis of symptom descriptions and drawings. AB - The specific aims of this study were to collect and analyze detailed symptom descriptions from patients with pediatric restless legs syndrome, ages 6 to 17 years, as well as assess symptom impact and the usefulness of drawings. Trained qualitative interviewers conducted face-to-face audio-recorded interviews of children and adolescents who met criteria for definite restless legs syndrome. Thirty-three patients in 3 age groups used 16 different categories of descriptors for restless legs sensations, with a mean of 3 or more categories used per patient in each age group. "Need to move/kick," "pain/hurts," "uncomfortable/cannot get comfortable," and "like bugs or ants/crawling" were the most common descriptors. Two-thirds reported daytime sensations, and nearly half had arm involvement. They described impact on sleep, cognitive function, and affect. Drawings provided useful diagnostic information. These detailed empirical data will be useful in clinical practice, as well as in the development of formal diagnostic tools and severity measures. PMID- 21636778 TI - RNA expression profiles from blood for the diagnosis of stroke and its causes. AB - A blood test to detect stroke and its causes would be particularly useful in babies, young children, and patients in intensive care units and for emergencies when imaging is difficult to obtain or is unavailable. Whole genome microarrays were used to show specific gene expression profiles in rats 24 hours after ischemic and hemorrhagic stroke, hypoxia, and hypoglycemia. These proof-of principle studies revealed that groups of genes (called gene profiles) can distinguish ischemic stroke patients from controls within 3 to 24 hours after the strokes. In addition, gene expression profiles have been developed that distinguish stroke due to large-vessel atherosclerosis from cardioembolic stroke. These profiles will be useful for predicting the causes of cryptogenic stroke. The results in adults suggest that similar diagnostic tools could be developed for children. PMID- 21636779 TI - Change in gross motor abilities of girls and women with rett syndrome over a 3- to 4-year period. AB - Rett syndrome is a rare but severe neurological disorder typically associated with a mutation in the MECP2 gene. We describe change in gross motor function over 3 to 4 years for 70 subjects participating in the Australian Rett Syndrome Database. Linear regression was used to assess relationships with age, genotype, and general and complex gross motor skills scores measured on the Gross Motor Scale for Rett syndrome. Skills were slightly better or maintained in approximately 40% of subjects and slightly decreased in approximately 60%. Teenagers and women who walked in 2004 were less likely to lose complex skills than those younger. Girls with a p.R294X mutation were more likely to lose complex motor skills, otherwise skill changes were spread across the mutation categories. In conclusion, small changes were observed over this period with greater stability of skills in teenagers and women with the ability to walk. PMID- 21636780 TI - Epilepsy treatment in Rett syndrome. AB - Rett syndrome is a neurodevelopmental disorder predominately affecting females. The majority of patients have epilepsy in the early stages of the disease. This study evaluates the clinical course of epilepsy and the effect of antiepileptic drug treatment in Rett syndrome using retrospective data analysis. Epilepsy was present in 16 of 19 (84%) patients with Rett syndrome in this series. The mean age of seizure onset was 4 years. Remission of seizures was achieved after the first monotherapy in 56% and after the second monotherapy in 18.5% of patients. Valproate, lamotrigine, and carbamazepine were the drugs used most frequently as monotherapy. Valproate monotherapy was highly effective as 75% of treated patients achieved seizure remission. Monotherapy with lamotrigine or carbamazepine was effective in half of the treated patients. There was a clear tendency toward seizure remission after the age of 15 years. PMID- 21636781 TI - Abnormal brain protein synthesis in language areas of children with pervasive developmental disorder: a L-[1-11C]-leucine PET study. AB - This study was performed to evaluate the cerebral protein synthesis rate of language brain regions in children with developmental delay with and without pervasive developmental disorder. The authors performed L-[1-(11)C]-leucine positron emission tomography (PET) on 8 developmental delay children with pervasive developmental disorder (mean age, 76.25 months) and 8 developmental delay children without pervasive developmental disorder (mean age, 77.63 months). They found a higher protein synthesis rate in developmental delay children with pervasive developmental disorder in the left posterior middle temporal region (P = .014). There was a significant correlation of the Gilliam Autism Rating Scale autism index score with the protein synthesis rate of the left posterior middle temporal region (r = .496, P = .05). In addition, significant asymmetric protein synthesis (right > left) was observed in developmental delay children without pervasive developmental disorder in the middle frontal and posterior middle temporal regions (P = .03 and P = .04, respectively). In conclusion, abnormal language area protein synthesis in developmentally delayed children may be related to pervasive symptoms. PMID- 21636782 TI - Long-term correlations in European socioeconomic conditions create a bias to conclusion that an invasion debt occurs. PMID- 21636783 TI - Structural basis for pregnenolone biosynthesis by the mitochondrial monooxygenase system. AB - In humans, the precursor to all steroid hormones, pregnenolone, is synthesized from cholesterol by an enzyme complex comprising adrenodoxin reductase (AdR), adrenodoxin (Adx), and a cytochrome P450 (P450scc or CYP11A1). This complex not only plays a key role in steroidogenesis, but also has long been a model to study electron transfer, multistep catalysis, and C-C bond cleavage performed by monooxygenases. Detailed mechanistic understanding of these processes has been hindered by a lack of structural information. Here we present the crystal structure of the complex of human Adx and CYP11A1--the first of a complex between a eukaryotic CYP and its redox partner. The structures with substrate and a series of reaction intermediates allow us to define the mechanism underlying sequential hydroxylations of the cholesterol and suggest the mechanism of C-C bond cleavage. In the complex the [2Fe-2S] cluster of Adx is positioned 17.4 A away from the heme iron of CYP11A1. This structure suggests that after an initial protein-protein association driven by electrostatic forces, the complex adopts an optimized geometry between the redox centers. Conservation of the interaction interface suggests that this mechanism is common for all mitochondrial P450s. PMID- 21636784 TI - Structure of the guanylyltransferase domain of human mRNA capping enzyme. AB - The enzyme guanylyltransferase (GTase) plays a central role in the three-step catalytic process of adding an (m7)GpppN cap cotranscriptionally to nascent mRNA (pre-mRNAs). The 5'-mRNA capping process is functionally and evolutionarily conserved from unicellular organisms to human. However, the GTases from viruses and yeast have low amino acid sequence identity (~25%) with GTases from mammals that, in contrast, are highly conserved (~98%). We have defined by limited proteolysis of human capping enzyme residues 229-567 as comprising the minimum enzymatically active human GTase (hGTase) domain and have determined the structure by X-ray crystallography. Seven related conformational states of hGTase exist in the crystal. The GTP-binding site is evolutionarily and structurally conserved. The positional variations of the oligonucleotide/oligosaccharide binding fold lid domain over the GTP-binding site provide snapshots of the opening and closing of the active site cleft through a swivel motion. The pattern of conserved surface residues in mammals, but not in yeast, supports the finding that the recognition of the capping apparatus by RNA polymerase II and associated transcription factors is highly conserved in mammals, and the mechanism may differ somewhat from that in yeast. The hGTase structure should help in the design of biochemical and molecular biology experiments to explore the proteinprotein and proteinRNA interactions that ensure regulated transcription of genes in humans and other mammals. PMID- 21636785 TI - MicroRNA-21 targets peroxisome proliferators-activated receptor-alpha in an autoregulatory loop to modulate flow-induced endothelial inflammation. AB - Adhesion of circulating monocytes to vascular endothelial cells (ECs) is a critical event leading to vascular inflammation and, hence, development of atherosclerosis. MicroRNAs (miRs) are a class of endogenous, highly conserved, noncoding small RNAs that play important roles in regulating gene expression and cellular function, as well as pathogenesis of atherosclerosis. Here, we showed that oscillatory shear stress (OSS) induces the expression of miR-21 at the transcriptional level in cultured human umbilical vein ECs via an increased binding of c-Jun, which is a component of transcription factor activator protein 1 (AP-1), to the promoter region of miR-21. OSS induction of miR-21 inhibited the translation, but not transcription, of peroxisome proliferators-activated receptor-alpha (PPARalpha) by 3'-UTR targeting. Overexpression of miR-21 up regulated AP-1 activation, which was attenuated by exogenous expression of PPARalpha. OSS and overexpression of miR-21 enhanced the expression of adhesion molecules vascular cell adhesion molecule-1 and monocyte chemotactic protein-1 and the consequential adhesion of monocytes to ECs. Overexpression of PPARalpha significantly attenuated the AP-1-mediated miR-21 expression. These results demonstrate a unique mechanism by which OSS induces AP-1-dependent miR-21 expression, which directly targets PPARalpha to inhibit its expression, thereby allowing activation of AP-1 and the promotion of monocyte adhesion. Our findings suggest the presence of a positive feedback loop that enables the sustained induction of miR-21, thus contributing to the proinflammatory responses of vascular endothelium under OSS. PMID- 21636786 TI - Increased Wingless (Wnt) signaling in pituitary progenitor/stem cells gives rise to pituitary tumors in mice and humans. AB - Wingless (Wnt)/beta-catenin signaling plays an essential role during normal development, is a critical regulator of stem cells, and has been associated with cancer in many tissues. Here we demonstrate that genetic expression of a degradation-resistant mutant form of beta-catenin in early Rathke's pouch (RP) progenitors leads to pituitary hyperplasia and severe disruption of the pituitary specific transcription factor 1-lineage differentiation resulting in extreme growth retardation and hypopituitarism. Mutant mice mostly die perinatally, but those that survive weaning develop lethal pituitary tumors, which closely resemble human adamantinomatous craniopharyngioma, an epithelial tumor associated with mutations in the human beta-catenin gene. The tumorigenic effect of mutant beta-catenin is observed only when expressed in undifferentiated RP progenitors, but tumors do not form when committed or differentiated cells are targeted to express this protein. Analysis of affected pituitaries indicates that expression of mutant beta-catenin leads to a significant increase in the total numbers of pituitary progenitor/stem cells as well as in their proliferation potential. Our findings provide insights into the role of the Wnt pathway in normal pituitary development and demonstrate a causative role for mutated beta-catenin in an undifferentiated RP progenitor in the genesis of murine and human craniopharyngioma. PMID- 21636787 TI - Intrinsic functional architecture predicts electrically evoked responses in the human brain. AB - Adaptive brain function is characterized by dynamic interactions within and between neuronal circuits, often occurring at the time scale of milliseconds. These complex interactions between adjacent and noncontiguous brain areas depend on a functional architecture that is maintained even in the absence of input. Functional MRI studies carried out during rest (R-fMRI) suggest that this architecture is represented in low-frequency (<0.1 Hz) spontaneous fluctuations in the blood oxygen level-dependent signal that are correlated within spatially distributed networks of brain areas. These networks, collectively referred to as the brain's intrinsic functional architecture, exhibit a remarkable correspondence with patterns of task-evoked coactivation as well as maps of anatomical connectivity. Despite this striking correspondence, there is no direct evidence that this intrinsic architecture forms the scaffold that gives rise to faster processes relevant to information processing and seizure spread. Here, we demonstrate that the spatial distribution and magnitude of temporally correlated low-frequency fluctuations observed with R-fMRI during rest predict the pattern and magnitude of corticocortical evoked potentials elicited within 500 ms after single-pulse electrical stimulation of the cerebral cortex with intracranial electrodes. Across individuals, this relationship was found to be independent of the specific regions and functional systems probed. Our findings bridge the immense divide between the temporal resolutions of these distinct measures of brain function and provide strong support for the idea that the low-frequency signal fluctuations observed with R-fMRI maintain and update the intrinsic architecture underlying the brain's repertoire of functional responses. PMID- 21636788 TI - Steroidogenic factor 1 directs programs regulating diet-induced thermogenesis and leptin action in the ventral medial hypothalamic nucleus. AB - The transcription factor steroidogenic factor 1 (SF-1) is exclusively expressed in the brain in the ventral medial hypothalamic nucleus (VMH) and is required for the development of this nucleus. However, the physiological importance of transcriptional programs regulated by SF-1 in the VMH is not well defined. To delineate the functional significance of SF-1 itself in the brain, we generated pre- and postnatal VMH-specific SF-1 KO mice. Both models of VMH-specific SF-1 KO were susceptible to high fat diet-induced obesity and displayed impaired thermogenesis after acute exposure to high fat diet. Furthermore, VMH-specific SF 1 KO mice showed significantly decreased LepR expression specifically in the VMH, leading to leptin resistance. Collectively, these results indicate that SF-1 directs transcriptional programs in the hypothalamus relevant to coordinated control of energy homeostasis, especially after excess caloric intake. PMID- 21636789 TI - Acidic nuclear phosphoprotein 32kDa (ANP32)B-deficient mouse reveals a hierarchy of ANP32 importance in mammalian development. AB - The highly conserved ANP32 proteins are proposed to function in a broad array of physiological activities through molecular mechanisms as diverse as phosphatase inhibition, chromatin regulation, caspase activation, and intracellular transport. On the basis of previous analyses of mice bearing targeted mutations of Anp32a or Anp32e, there has been speculation that all ANP32 proteins play redundant roles and are dispensable for normal development. However, more recent work has suggested that ANP32B may in fact have functions that are not shared by other ANP32 family members. Here we report that ANP32B expression is associated with a poor prognosis in human breast cancer, consistent with the increased levels of Anp32b mRNA present in proliferating wild-type (WT) murine embryonic fibroblasts and stimulated WT B and T lymphocytes. Moreover, we show that, contrary to previous assumptions, Anp32b is very important for murine embryogenesis. In a mixed genetic background, ANP32B-deficient mice displayed a partially penetrant perinatal lethality that became fully penetrant in a pure C57BL/6 background. Surviving ANP32B-deficient mice showed reduced viability due to variable defects in various organ systems. Study of compound mutants lacking ANP32A, ANP32B, and/or ANP32E revealed previously hidden roles for ANP32A in mouse development that became apparent only in the complete absence of ANP32B. Our data demonstrate a hierarchy of importance for the mammalian Anp32 genes, with Anp32b being the most critical for normal development. PMID- 21636790 TI - Apoptosis and the selective survival of host animals following thermal bleaching in zooxanthellate corals. AB - During the past several decades, numerous reports from disparate geographical areas have documented an increased frequency of "bleaching" in reef-forming corals. The phenomenon, triggered by increased sea surface temperatures, occurs when the cnidarian hosts digest and/or expel their intracellular, photosynthetic dinoflagellate symbionts ("zooxanthellae" in the genus Symbiodinium). Although coral bleaching is often followed by the death of the animal hosts, in some cases, the animal survives and can be repopulated with viable zooxanthellae. The physiological factors determining the ability of the coral to survive bleaching events are poorly understood. In this study, we experimentally established that bleaching and death of the host animal involve a caspase-mediated apoptotic cascade induced by reactive oxygen species produced primarily by the algal symbionts. In addition, we demonstrate that, although some corals naturally suppress caspase activity and significantly reduce caspase concentration under high temperatures as a mechanism to prevent colony death from apoptosis, even sensitive corals can be prevented from dying by application of exogenous inhibitors of caspases. Our results indicate that variability in response to thermal stress in corals is determined by a four-element, combinatorial genetic matrix intrinsic to the specific symbiotic association. Based on our experimental data, we present a working model in which the phenotypic expression of this symbiont/host relationship places a selective pressure on the symbiotic association. The model predicts the survival of the host animals in which the caspase-mediated apoptotic cascade is down-regulated. PMID- 21636791 TI - Algal ice-binding proteins change the structure of sea ice. PMID- 21636792 TI - Intelligence differences may explain the link between childhood psychological problems and adult socioeconomic status. PMID- 21636793 TI - Comparison of direct action of thiazolidinediones and glucocorticoids on renal podocytes: protection from injury and molecular effects. AB - The U.S. Food and Drug Administration-approved thiazolidinediones pioglitazone and rosiglitazone are peroxisome proliferator-activated receptor-gamma (PPARgamma) agonists developed to control serum glucose in patients with diabetes. They have been found to reduce proteinuria and microalbuminuria in both diabetic nephropathy and nondiabetic glomerulosclerosis. We hypothesized that the renal protective effects of thiazolidinediones result, at least in part, from their direct action on podocytes, similar to glucocorticoids. Treatment with pioglitazone, rosiglitazone, or dexamethasone significantly protected podocytes against puromycin aminonucleoside-induced injury (designed to mimic nephrotic syndrome-related injury), as determined by both cell survival and actin cytoskeletal integrity. Furthermore, we compared the ability of these drugs to modulate key signaling pathways in podocytes that may be critical to their protective effects. Rosiglitazone deactivated the mitogen-activated protein kinases (MAPKs), extracellular signal-regulated kinases 1/2, p38 MAPK, and stress activated protein kinase/c-Jun NH2-terminal kinase, whereas pioglitazone did not, and dexamethasone deactivated to some extent. Similar to dexamethasone, both thiazolidinediones increased the glucocorticoid receptor phosphorylation, and this response to rosiglitazone and possibly to pioglitazone was PPARgamma dependent. Furthermore, both drugs mimicked or enhanced the effects of dexamethasone on glucocorticoid-responsive genes in a PPARgamma- and glucocorticoid receptor-dependent manner. In addition, both thiazolidinediones mimicked dexamethasone-induced effects on calcineurin activity. In summary, thiazolidinediones are able to modulate the glucocorticoid pathway and exert direct protective effects on podocytes, similar to glucocorticoids. This suggests that thiazolidinediones may have potential clinical utility as either primary or adjunctive therapy for nephrotic syndrome or other diseases treated with glucocorticoids. These findings may also lend mechanistic insight into the well established but poorly understood renal protective effects of thiazolidinediones in diabetic nephropathy. PMID- 21636794 TI - Skin intrinsic fluorescence is associated with hemoglobin A(1c )and hemoglobin glycation index but not mean blood glucose in children with type 1 diabetes. AB - OBJECTIVE: To evaluate the relationship between skin advanced glycation end products (sAGEs) with mean blood glucose (MBG), hemoglobin A(1c) (HbA(1c)), and MBG-independent, between-patient differences in HbA(1c) among children with type 1 diabetes. RESEARCH DESIGN AND METHODS: Children aged 5 to 20 years with type 1 diabetes of at least 1 year duration participated. At a clinic visit, sAGE was estimated noninvasively by measurement of skin intrinsic fluorescence (SIF). SIF data were adjusted to correct for variation in skin pigmentation. MBG independent, between-patient differences in HbA(1c) were examined by statistically controlling HbA(1c) for MBG or alternatively by use of a hemoglobin glycation index (HGI). Results were similar whether HbA(1c), MBG, and HGI were analyzed as single values from the time of the SIF examination visit or as the mean values from all available visits of the patient. RESULTS: HbA(1c) was correlated with MBG (r = 0.5; P < 0.001; n = 110). HbA(1c) and HGI, but not MBG, were statistically associated with SIF after adjustment for age, duration of diabetes, race, sex, and BMI z-score. SIF increased with age and duration of diabetes and was higher in girls than boys. CONCLUSIONS: sAGE levels estimated by SIF increase with age, duration of diabetes, and female sex. sAGE is correlated with MBG-independent biological variation in HbA(1c), but not with MBG itself. These results suggest that factors besides MBG that influence HbA(1c) levels also contribute to accumulation of sAGE. PMID- 21636795 TI - Correlates of quality of life in older adults with diabetes: the diabetes & aging study. AB - OBJECTIVE: To evaluate associations between health-related quality of life (HRQL) and geriatric syndromes, diabetes complications, and hypoglycemia in older adults with diabetes. RESEARCH DESIGN AND METHODS: A race-stratified random sample of 6,317 adults with type 2 or type 1 diabetes, aged 60 to 75 years, enrolled in Kaiser Permanente Northern California, who completed a survey that included a HRQL instrument based on the Short Form 8-item health survey. Administrative records were used to ascertain diagnoses of geriatric syndromes, diabetes complications, and hypoglycemia. Associations were estimated between HRQL and exposures in exposure-specific and combined exposure models (any syndrome, any complication, or hypoglycemia). Conservatively, differences of >=3 points were considered the minimally important difference in HRQL scores. RESULTS: HRQL was lower with nearly all exposures of interest. The lowest physical HRQL was associated with amputation. In combined exposure models, geriatric syndromes ( 5.3 [95% CI -5.8 to -4.8], P < 0.001) and diabetes complications (-3.5 [-4.0 to 2.9], P < 0.001) were associated with lower physical HRQL. The lowest mental HRQL was associated with depression, underweight (BMI <18 kg/m(2)), amputation, and hypoglycemia. In combined exposure models, only hypoglycemia was associated with lower mental HRQL (-4.0 [-7.0 to -1.1], P = 0.008). CONCLUSIONS: Geriatric syndromes and hypoglycemia are associated with lower HRQL to a comparable degree as diabetes complications. Addressing geriatric syndromes and avoiding hypoglycemia should be given as high a priority as preventing diabetes complications in older adults with diabetes. PMID- 21636796 TI - Effect of nurse case management compared with usual care on controlling cardiovascular risk factors in patients with diabetes: a randomized controlled trial. AB - OBJECTIVE: To determine whether nurse case management with a therapeutic algorithm could effectively improve rates of control for hypertension, hyperglycemia, and hyperlipidemia compared with usual care among veterans with diabetes. RESEARCH DESIGN AND METHODS: A randomized controlled trial of diabetic patients that had blood pressure (BP) >140/90 mmHg, hemoglobin A(1c) (HbA(1c)) >9.0%, or LDL >100 mg/dL. Intervention patients received case management (n = 278) versus usual care (n = 278) over a 1-year period. The primary outcome was the percentage of patients achieving simultaneous control of all three parameters (defined by BP <130/80 mmHg, HbA(1c) <8.0%, and LDL <100 mg/dL) at 1 year. Secondary outcomes included improvements within each individual component of the composite primary outcome. Differences between groups were analyzed using t tests, Pearson chi(2) tests, and linear and logistic regression. RESULTS: A greater number of individuals assigned to case management achieved the primary study outcome of having all three outcome measures under control (61 [21.9%] compared with 28 [10.1%] in the usual care group [P < 0.01]). In addition, a greater number of individuals assigned to the intervention group achieved the individual treatment goals of HbA(1c) <8.0% (73.7 vs. 65.8%, P = 0.04) and BP <130/80 mmHg (45.0 vs. 25.4%, P < 0.01), but not for LDL <100 mg/dL (57.6 vs. 55.4%, P = 0.61), compared with those in the usual care group. CONCLUSIONS: In patients with diabetes, nurse case managers using a treatment algorithm can effectively improve the number of individuals with control of multiple cardiovascular risk factors at 1 year. PMID- 21636797 TI - Evolution of renal hyperfiltration and arterial stiffness from adolescence into early adulthood in type 1 diabetes. AB - OBJECTIVE: To determine, in a small but carefully physiologically characterized cohort of subjects with uncomplicated type 1 diabetes, the changes in renal hemodynamic function and arterial stiffness that occur over time as the participants transitioned from adolescence into early adulthood. The classical paradigm for type 1 diabetes suggests that glomerular filtration rate (GFR) declines in patients with renal hyperfiltration, but the natural history of peripheral vascular function abnormalities in uncomplicated type 1 diabetes is less well understood, particularly as patients transition from adolescence to early adulthood. RESEARCH DESIGN AND METHODS: Renal hemodynamic function (inulin and p-aminohippuric acid clearance), blood pressure, arterial stiffness (radial augmentation index), albumin excretion, and circulating renin-angiotensin system measures were obtained during clamped euglycemia at baseline and at follow-up 6.8 +/- 2.5 years later in 10 patients with hyperfiltration (GFR >=135 mL/min/1.73 m(2)) and in 8 with normofiltration. RESULTS: Compared with baseline values, GFR (171 +/- 20 to 120 +/- 15 mL/min/1.73 m(2)) and filtration fraction (FF, 0.24 +/- 0.06 to 0.18 +/- 0.03) declined in hyperfilterers (ANOVA P <= 0.033), and renal vascular resistance increased (0.0678 +/- 0.0135 to 0.0783 +/- 0.0121 mmHg/L/min, P = 0.017). GFR and FF did not change in normofiltering subjects. In contrast, the radial augmentation index decreased in hyperfiltering (1.2 +/- 11.7 to -11.0 +/- 7.8%) and normofiltering (14.3 +/- 14.0 to 2.5 +/- 14.6%) subjects (within group changes, ANOVA P <= 0.030). The decline in circulating aldosterone levels was similar in both groups. CONCLUSIONS: During the transition from adolescence to early adulthood, hyperfiltration is not sustained in subjects with type 1 diabetes, whereas GFR remains stable in normofiltering subjects. Our findings suggest early normofiltration may predict stable renal function. In contrast, arterial stiffness decreased in all patients regardless of filtration status, suggesting that age-related increases in arterial stiffness occur at older ages. PMID- 21636798 TI - Optimal glycemic control, pre-eclampsia, and gestational hypertension in women with type 1 diabetes in the diabetes and pre-eclampsia intervention trial. AB - OBJECTIVE: To assess the relationship between glycemic control, pre-eclampsia, and gestational hypertension in women with type 1 diabetes. RESEARCH DESIGN AND METHODS: Pregnancy outcome (pre-eclampsia or gestational hypertension) was assessed prospectively in 749 women from the randomized controlled Diabetes and Pre-eclampsia Intervention Trial (DAPIT). HbA(1c) (A1C) values were available up to 6 months before pregnancy (n = 542), at the first antenatal visit (median 9 weeks) (n = 721), at 26 weeks' gestation (n = 592), and at 34 weeks' gestation (n = 519) and were categorized as optimal (<6.1%: referent), good (6.1-6.9%), moderate (7.0-7.9%), and poor (>=8.0%) glycemic control, respectively. RESULTS: Pre-eclampsia and gestational hypertension developed in 17 and 11% of pregnancies, respectively. Women who developed pre-eclampsia had significantly higher A1C values before and during pregnancy compared with women who did not develop pre-eclampsia (P < 0.05, respectively). In early pregnancy, A1C >= 8.0% was associated with a significantly increased risk of pre-eclampsia (odds ratio 3.68 [95% CI 1.17-11.6]) compared with optimal control. At 26 weeks' gestation, A1C values >= 6.1% (good: 2.09 [1.03-4.21]; moderate: 3.20 [1.47-7.00]; and poor: 3.81 [1.30-11.1]) and at 34 weeks' gestation A1C values >= 7.0% (moderate: 3.27 [1.31-8.20] and poor: 8.01 [2.04-31.5]) significantly increased the risk of pre eclampsia compared with optimal control. The adjusted odds ratios for pre eclampsia for each 1% decrement in A1C before pregnancy, at the first antenatal visit, at 26 weeks' gestation, and at 34 weeks' gestation were 0.88 (0.75-1.03), 0.75 (0.64-0.88), 0.57 (0.42-0.78), and 0.47 (0.31-0.70), respectively. Glycemic control was not significantly associated with gestational hypertension. CONCLUSIONS: Women who developed pre-eclampsia had significantly higher A1C values before and during pregnancy. These data suggest that optimal glycemic control both early and throughout pregnancy may reduce the risk of pre-eclampsia in women with type 1 diabetes. PMID- 21636799 TI - Estimating the risk of developing type 2 diabetes: a comparison of several risk scores: the Cohorte Lausannoise study. AB - OBJECTIVE: To compare in the Swiss population the results of several scores estimating the risk of developing type 2 diabetes. RESEARCH DESIGN AND METHODS: This was a single-center, cross-sectional study conducted between 2003 and 2006 in Lausanne, Switzerland. Overall, 3,251 women and 2,937 men, aged 35-75 years, were assessed, of which 5,760 (93%) were free from diabetes and included in the current study. The risk of developing type 2 diabetes was assessed using seven different risk scores, including clinical data with or without biological data. Participants were considered to be eligible for primary prevention according to the thresholds provided for each score. The results were then extrapolated to the Swiss population of the same sex and age. RESULTS: The risk of developing type 2 diabetes increased with age in all scores. The prevalence of participants at high risk ranged between 1.6 and 24.9% in men and between 1.1 and 15.7% in women. Extrapolated to the Swiss population of similar age, the overall number of participants at risk, and thus susceptible to intervention, ranged between 46,708 and 636,841. In addition, scores that included the same clinical variables led to a significantly different prevalence of participants at risk (4.2% [95% CI 3.4 5.0] vs. 12.8% [11.5-14.1] in men and 2.9% [2.4-3.6] vs. 6.0% [5.2-6.9] in women). CONCLUSIONS; The prevalence of participants at risk for developing type 2 diabetes varies considerably according to the scoring system used. To adequately prevent type 2 diabetes, risk-scoring systems must be validated for each population considered. PMID- 21636800 TI - Etiological approach to characterization of diabetes type: the SEARCH for Diabetes in Youth Study. AB - OBJECTIVE: To describe an etiologic approach to classification of diabetes types in youth based on the 1997 American Diabetes Association (ADA) framework, using data from the SEARCH for Diabetes in Youth Study. RESEARCH DESIGN AND METHODS: SEARCH conducted a comprehensive assessment of 2,291 subjects aged <20 years with recently diagnosed diabetes. Using autoimmunity (at least one of two diabetes autoantibodies) and insulin sensitivity (equation validated against hyperinsulinemic-euglycemic clamps) as the main etiologic markers, we described four categories along a bidimensional spectrum: autoimmune plus insulin-sensitive (IS), autoimmune plus insulin-resistant (IR), nonautoimmune plus IS, and nonautoimmune plus IR. We then explored how characteristics, including genetic susceptibility to autoimmunity (HLA genotypes), insulin deficiency, and clinical factors varied across these four categories. RESULTS: Most subjects fell into either the autoimmune plus IS (54.5%) or nonautoimmune plus IR categories (15.9%) and had characteristics that align with traditional descriptions of type 1 or type 2 diabetes. The group classified as autoimmune plus IR (19.5%) had similar prevalence and titers of diabetes autoantibodies and similar distribution of HLA risk genotypes to those in the autoimmune plus IS group, suggesting that it includes individuals with type 1 diabetes who are obese. The group classified as nonautoimmune plus IS (10.1%) likely includes individuals with undetected autoimmunity but may also include those with monogenic diabetes and thus requires further testing. CONCLUSIONS: The SEARCH study offers researchers and clinicians a practical application for the etiologic classification of diabetes type and at the same time identifies a group of youths who would benefit from further testing. PMID- 21636801 TI - Bacterial endotoxin activity in human serum is associated with dyslipidemia, insulin resistance, obesity, and chronic inflammation. AB - OBJECTIVE: To investigate whether bacterial lipopolysaccharide (LPS) activity in human serum is associated with the components of the metabolic syndrome (MetS) in type 1 diabetic patients with various degrees of kidney disease and patients with IgA glomerulonephritis (IgAGN). RESEARCH DESIGN AND METHODS: Serum LPS activity was determined with the Limulus Amoebocyte Lysate chromogenic end point assay in type 1 diabetic patients with a normal albumin excretion rate (n = 587), microalbuminuria (n = 144), macroalbuminuria (n = 173); patients with IgAGN (n = 98); and in nondiabetic control subjects (n = 345). The relationships of the LPS/HDL ratio and MetS-associated variables were evaluated with Pearson correlation. RESULTS: The MetS was more prevalent in type 1 diabetic patients (48%) than in patients with IgAGN (15%). Diabetic patients with macroalbuminuria had a significantly higher serum LPS/HDL ratio than patients with IgAGN. In the normoalbuminuric type 1 diabetic group, patients in the highest LPS/HDL quartile were diagnosed as having the MetS three times more frequently than patients in the lowest quartile (69 vs. 22%; P < 0.001). High LPS activity was associated with higher serum triglyceride concentration, earlier onset of diabetes, increased diastolic blood pressure, and elevated urinary excretion of monocyte chemoattractant protein-1. CONCLUSIONS: High serum LPS activity is strongly associated with the components of the MetS. Diabetic patients with kidney disease seem to be more susceptible to metabolic endotoxemia than patients with IgAGN. Bacterial endotoxins may thus play an important role in the development of the metabolic and vascular abnormalities commonly seen in obesity and diabetes related diseases. PMID- 21636802 TI - Placental growth factor regulates cardiac adaptation and hypertrophy through a paracrine mechanism. AB - RATIONALE: Paracrine growth factor-mediated crosstalk between cardiac myocytes and nonmyocytes in the heart is critical for programming adaptive cardiac hypertrophy in which myocyte size, capillary density, and the extracellular matrix function coordinately. OBJECTIVE: To examine the role that placental growth factor (PGF) plays in the heart as a paracrine regulator of cardiac adaptation to stress stimulation. METHODS AND RESULTS: PGF is induced in the heart after pressure-overload stimulation, where it is expressed in both myocytes and nonmyocytes. We generated cardiac-specific and adult inducible PGF overexpressing transgenic mice and analyzed Pgf(-/-) mice to examine the role that this factor plays in cardiac disease and paracrine signaling. Although PGF transgenic mice did not have a baseline phenotype or a change in capillary density, they did exhibit a greater cardiac hypertrophic response, a greater increase in capillary density, and increased fibroblast content in the heart in response to pressure-overload stimulation. PGF transgenic mice showed a more adaptive type of cardiac growth that was protective against signs of failure with pressure overload and neuroendocrine stimulation. Antithetically, Pgf(-/-) mice rapidly died of heart failure within 1 week of pressure overload, they showed an inability to upregulate angiogenesis, and they showed significantly less fibroblast activity in the heart. Mechanistically, we show that PGF does not have a direct effect on cardiomyocytes but works through endothelial cells and fibroblasts by inducing capillary growth and fibroblast proliferation, which secondarily support greater cardiac hypertrophy through intermediate paracrine growth factors such as interleukin-6. CONCLUSIONS: PGF is a secreted factor that supports hypertrophy and cardiac function during pressure overload by affecting endothelial cells and fibroblasts that in turn stimulate and support the myocytes through additional paracrine factors. PMID- 21636803 TI - Overexpression of VEGF165b, an inhibitory splice variant of vascular endothelial growth factor, leads to insufficient angiogenesis in patients with systemic sclerosis. AB - RATIONALE: Systemic sclerosis (SSc) is characterized by widespread microangiopathy, fibrosis, and autoimmunity. Despite the lack of angiogenesis, the expression of vascular endothelial growth factor A (VEGF) was shown to be upregulated in SSc skin and circulation; however, previous studies did not distinguish between proangiogenic VEGF(165) and antiangiogenic VEGF(165)b isoforms, which are generated by alternative splicing in the terminal exon of VEGF pre-RNA. OBJECTIVE: We investigated whether VEGF isoform expression could be altered in skin and circulation of patients with SSc. METHODS AND RESULTS: Here, we show that the endogenous antiangiogenic VEGF(165)b splice variant is selectively overexpressed at both the mRNA and protein levels in SSc skin. Elevated VEGF(165)b expression correlated with increased expression of profibrotic transforming growth factor-beta1 and serine/arginine protein 55 splicing factor in keratinocytes, fibroblasts, endothelial cells, and perivascular inflammatory cells. Circulating levels of VEGF(165)b were significantly higher in patients with SSc than in control subjects. Microvascular endothelial cells (MVECs) isolated from SSc skin expressed and released higher levels of VEGF(165)b than healthy MVECs. Transforming growth factor-beta1 upregulated the expression of VEGF(165)b and serine/arginine protein 55 in both SSc and healthy MVECs. In SSc MVECs, VEGF receptor-2 was overexpressed, but its phosphorylation was impaired. Recombinant VEGF(165)b and SSc-MVEC-conditioned medium inhibited VEGF(165)-mediated VEGF receptor-2 phosphorylation and capillary morphogenesis in healthy MVECs. The addition of anti-VEGF(165)b blocking antibodies abrogated the antiangiogenic effect of SSc-MVEC-conditioned medium. Capillary morphogenesis was severely impaired in SSc MVECs and could be ameliorated by treatment with recombinant VEGF(165) and anti-VEGF(165)b blocking antibodies. CONCLUSIONS: In SSc, a switch from proangiogenic to antiangiogenic VEGF isoforms may have a crucial role in the insufficient angiogenic response to chronic ischemia. PMID- 21636804 TI - Thioredoxin-interacting protein mediates TRX1 translocation to the plasma membrane in response to tumor necrosis factor-alpha: a key mechanism for vascular endothelial growth factor receptor-2 transactivation by reactive oxygen species. AB - OBJECTIVE: Thioredoxin-interacting protein (TXNIP) promotes inflammation in endothelial cells (EC) by binding to thioredoxin-1 (TRX1) in a redox-dependent manner. Formation of the TXNIP-TRX1 complex relieves inhibition of the apoptosis signal-regulating kinase 1-c-Jun N-terminal kinase-vascular cell adhesion molecule-1 pathway. Because TXNIP is an alpha-arrestin with numerous protein protein interacting domains, we hypothesized that TXNIP-TRX1 trafficking should alter function of EC exposed to reactive oxygen species (ROS). METHODS AND RESULTS: In response to physiological levels of ROS (10 ng/mL tumor necrosis factor-alpha and 30 MUmol/L H(2)O(2)), TXNIP-TRX1 translocated to the plasma membrane in human umbilical vein EC, with a peak at 30 minutes, as measured by immunofluorescence colocalization with vascular endothelial-cadherin, cell fractionation, and membrane sheet assay. TXNIP-mediated translocation of TRX1 to the membrane required TXNIP and TRX1 binding, as evidenced by inability of the ROS-insensitive TXNIP-Cys247Ser mutant to promote membrane localization. Vascular endothelial growth factor signaling required TXNIP, as shown by significant decreases in plasma membrane tyrosine phosphorylation and EC migration after TRX1 knockdown. Furthermore, TXNIP knockdown increased human umbilical vein EC apoptosis induced by tumor necrosis factor. Rescue with TXNIP-wild-type but not TXNIP-Cys247Ser prevented cell death. CONCLUSIONS: These findings suggest a novel role for the TXNIP-TRX1 complex to enable inflammation by promoting EC survival and vascular endothelial growth factor signaling under conditions of physiological oxidative stress. PMID- 21636805 TI - Response gene to complement 32 promotes vascular lesion formation through stimulation of smooth muscle cell proliferation and migration. AB - OBJECTIVE: The objectives of this study were to determine the role of response gene to complement 32 (RGC-32) in vascular lesion formation after experimental angioplasty and to explore the underlying mechanisms. METHODS AND RESULTS: Using a rat carotid artery balloon-injury model, we documented for the first time that neointima formation was closely associated with a significantly increased expression of RGC-32 protein. Short hairpin RNA knockdown of RGC-32 via adenovirus-mediated gene delivery dramatically inhibited the lesion formation by 62% as compared with control groups 14 days after injury. Conversely, RGC-32 overexpression significantly promoted the neointima formation by 33%. Gain- and loss-of-function studies in primary culture of rat aortic smooth muscle cells (RASMCs) indicated that RGC-32 is essential for both the proliferation and migration of RASMCs. RGC-32 induced RASMC proliferation by enhancing p34(CDC2) activity. RGC-32 stimulated the migration of RASMC by inducing focal adhesion contact and stress fiber formation. These effects were caused by the enhanced rho kinase II-alpha activity due to RGC-32-induced downregulation of Rad GTPase. CONCLUSIONS: RGC-32 plays an important role in vascular lesion formation following vascular injury. Increased RGC-32 expression in vascular injury appears to be a novel mechanism underlying the migration and proliferation of vascular smooth muscle cells. Therefore, targeting RGC-32 is a potential therapeutic strategy for the prevention of vascular remodeling in proliferative vascular diseases. PMID- 21636806 TI - Targeted disruption of the prostaglandin E2 E-prostanoid 2 receptor exacerbates vascular neointimal formation in mice. AB - OBJECTIVE: Restenosis after angioplasty remains a major clinical problem. Prostaglandin E(2) (PGE(2)) plays an important role in vascular homeostasis. The PGE(2) receptor E-prostanoid 2 (EP2) is involved in the proliferation and migration of various cell types. We aimed to determine the role of EP2 in the pathogenesis of neointimal formation after vascular injury. METHODS AND RESULTS: Wire-mediated vascular injury was induced in the femoral arteries of male wild type (EP2+/+) and EP2 gene-deficient (EP2-/-) mice. In EP2+/+ mice, EP2 mRNA expression was increased in injured vessels for at least 4 weeks after vascular injury. Neointimal hyperplasia was markedly accelerated in EP2-/- mice, which was associated with increased proliferation and migration of vascular smooth muscle cells (VSMCs) and increased cyclin D1 expression in the neointima layer. Platelet derived growth factor-BB (PDGF-BB) treatment resulted in more significant cell proliferation and migration in VSMCs of EP2-/- mice than in those of EP2+/+ mice. Activation and overexpression of EP2 attenuated PDGF-BB-elicited cell proliferation and migration, induced G(1)->S-phase arrest and reduced PDGF-BB stimulated extracellular signal-regulated kinase phosphorylation in EP2+/+ VSMCs. CONCLUSIONS: These findings reveal a novel role of the EP2 receptor in neointimal hyperplasia after arterial injury. The EP2 receptor may represent a potential therapeutic target for restenosis after angioplasty. PMID- 21636807 TI - Dietary phosphate modulates atherogenesis and insulin resistance in apolipoprotein E knockout mice--brief report. AB - OBJECTIVE: Epidemiological studies link higher serum phosphate and the phosphatonin fibroblast growth factor 23 with cardiovascular events and atheroma, and they link lower serum phosphate with insulin resistance and the metabolic syndrome. We investigated whether manipulating dietary phosphate influences atherogenesis or insulin sensitivity in mice. METHODS AND RESULTS: Apolipoprotein E knockout mice were fed an atherogenic diet with low (0.2%), standard (0.6%), or high (1.6%) phosphate content. Serum phosphate and fibroblast growth factor 23 significantly increased with increasing dietary phosphate intake, but lipid profile and blood pressure were unaffected. After 20 weeks, mice on the higher phosphate diet had significantly more atheroma at the aortic sinus (42+/-1.9% versus 30+/-1.5% for high versus low phosphate, P<0.01). Compared with standard and high-phosphate diet groups, mice on a low-phosphate diet had more adipose tissue and a 4-fold increase in insulin resistance measured by homeostatic model assessment (43.7+/-9.3 versus 8.9+/-0.7 for low versus high phosphate, P<0.005). CONCLUSIONS: A high-phosphate diet accelerates atherogenesis in apolipoprotein E( /-) mice, whereas low phosphate intake induces insulin resistance. These data indicate for the first time that controlling dietary phosphate intake may influence development of both atherosclerosis and the metabolic syndrome. PMID- 21636808 TI - Anoxia-reoxygenation enhances platelet thromboxane A2 production via reactive oxygen species-generated NOX2: effect in patients undergoing elective percutaneous coronary intervention. AB - OBJECTIVE: Platelets undergoing anoxia-reoxygenation (AR) simultaneously increase reactive oxygen species (ROS) and thromboxane (Tx) B(2). Our aim was to assess whether there is an interplay between activation of NOX2, the catalytic subunit of NADPH oxidase, and platelet TxB(2) in vitro and in vivo. METHODS AND RESULTS: Platelets that underwent AR had enhanced ROS. This was associated with NOX2 activation and was inhibited by incubation with NOX2-blocking peptide. AR was associated with TxB(2) and isoprostane production, which were inhibited by NOX2 blocking peptide, vitamin C, and the inhibitor of phospholipase A(2). Platelet incubation with 100 MUmol/L aspirin fully prevented AR-induced TxA(2) but did not affect isoprostane production. We included 56 aspirin-treated patients undergoing elective percutaneous coronary intervention (PCI) who were randomly allocated to receive either placebo or intravenous infusion of 1 g of vitamin C. Blood TxB(2), isoprostanes, and soluble NOX2-derived peptide, a marker of systemic NADPH oxidase activation, significantly increased at 60 and 120 minutes after PCI in placebo-treated but not in vitamin C-treated patients. CONCLUSIONS: AR is associated with overproduction of platelet TxB(2) and isoprostanes, which is dependent on NOX2-dependent ROS generation. Low doses of aspirin are unable to prevent TxB(2) formation in patients who undergo PCI. PMID- 21636809 TI - Human aldose reductase expression accelerates atherosclerosis in diabetic apolipoprotein E-/- mice. AB - OBJECTIVE: There are several pathways that mediate the aberrant metabolism of glucose and that might induce greater vascular damage in the setting of diabetes. The polyol pathway mediated by aldose reductase (AR) has been postulated to be one such pathway. However, it has been reported that AR reduces toxic lipid aldehydes and, under some circumstances, might be antiatherogenic. METHODS AND RESULTS: Atherosclerosis development was quantified in 2 lines of transgenic mice expressing human AR (hAR) crossed on the apolipoprotein E knockout background. The transgenes were used to increase the normally low levels of this enzyme in wild-type mice. Both generalized hAR overexpression and hAR expression via the Tie 2 promoter increased lesion size in streptozotocin diabetic mice. In addition, pharmacological inhibition of AR reduced lesion size. CONCLUSIONS: Although in some settings AR expression might reduce levels of toxic aldehydes, transgenic expression of this enzyme within the artery wall leads to greater atherosclerosis. PMID- 21636810 TI - Entrainment for distinguishing atypical atrioventricular node reentrant tachycardia from atrioventricular reentrant tachycardia over septal accessory pathways with long-RP [corrected] tachycardia. AB - BACKGROUND: The response to right ventricular (RV) entrainment is useful to distinguish atypical AV node reentrant tachycardia from AV reentrant tachycardia using a septal accessory pathway. Whether entrainment can differentiate between AV node reentrant tachycardia and AV reentrant tachycardia in patients with long RP tachycardia has not been systematically validated. METHODS AND RESULTS: Twenty four patients with concealed septal accessory pathways who had an electrophysiology study between January 1, 2000, and January 1, 2010, were included (age, 38 +/- 17 years; men, 17). Entrainment was performed from the RV apex pacing at cycle length 20 to 40 ms shorter than tachycardia cycle length (TCL). The mean TCL was 390 +/- 80 ms, the mean AH interval during tachycardia was 151 +/- 57 ms, and the mean ventriculoatrial (VA) time was 182 +/- 103 ms. Twelve patients had typical accessory pathways (VA/TCL <40%), and 12 had slowly conducting accessory pathways (VA/TCL >= 40%). In all patients with typical accessory pathways, the postpacing interval minus the TCL (PPI-TCL) was <115 ms and the difference in the VA interval during pacing and tachycardia (StimA-VA) was <85 ms. On the other hand, in 6 of the 12 patients in the slowly conducting group, the PPI-TCL was >115 ms, and the StimA-VA was > 85 ms. CONCLUSIONS: Slowly conducting accessory pathways frequently yield RV entrainment criteria traditionally attributable to AV node reentry. Distinguishing AV node reentry from AV reentry in patients with long-RP tachycardia requires other criteria. PMID- 21636811 TI - White matter hyperintensity penumbra. AB - BACKGROUND AND PURPOSE: White matter hyperintensities (WMHs) are associated with progressive age-related cognitive decline and cardiovascular risk factors, but their biological relevance as indicators of generalized white matter injury is unclear. Diffusion tensor imaging provides more sensitive indications of subtle white matter disruption and can therefore clarify whether WMHs represent foci of generalized white matter damage that extends over a broader neighborhood. METHODS: Two hundred eight participants from the University of California, Davis Alzheimer's Disease Center received a comprehensive clinical evaluation and brain MRI including fluid-attenuated inversion recovery and diffusion tensor imaging sequences. Voxelwise maps of WMHs were produced from fluid-attenuated inversion recovery using a standardized WMH detection protocol. Fractional anisotropy maps were calculated from diffusion tensor imaging. All WMH and fractional anisotropy maps were coregistered to a standardized space. For each normal-appearing white matter voxel in each subject fluid-attenuated inversion recovery scan, a neighborhood white matter injury score was calculated that increased with increasing number and proximity of WMH in the vicinity of the normal-appearing white matter voxel. Fractional anisotropy was related to neighborhood white matter injury using a nonlinear mixed effect model controlling for relevant confounding factors. RESULTS: Fractional anisotropy was found to decrease as neighborhood white matter injury increased (beta = -0.0017/%, P < 0.0001) with an accelerated rate (P < 0.0001) for neighborhood white matter injury >0.4. An increase of 1% in neighborhood white matter injury score was associated with a decrease in mean fractional anisotropy of 0.012 (P < 0.001). CONCLUSIONS: WMH may represent foci of more widespread and subtle white matter changes rather than distinct, sharply delineated anatomic abnormalities. We use the term white matter hyperintensities penumbra to explain this phenomenon. PMID- 21636812 TI - U.S. Wingspan Registry: 12-month follow-up results. AB - BACKGROUND AND PURPOSE: The purpose of this study is to present 12-month follow up results for a series of patients undergoing percutaneous transluminal angioplasty and stenting with the Gateway-Wingspan stenting system (Boston Scientific) for the treatment of symptomatic intracranial atherostenosis. METHODS: Clinical and angiographic follow-up results were recorded for patients from 5 participating institutions. Primary end points were stroke or death within 30 days of the stenting procedure or ipsilateral stroke after 30 days. RESULTS: During a 21-month study period, 158 patients with 168 intracranial atherostenotic lesions (50% to 99%) were treated with the Gateway-Wingspan system. The average follow-up duration was 14.2 months with 143 patients having at least 3 months of clinical follow-up and 110 having at least 12 months. The cumulative rate of the primary end point was 15.7% for all patients and 13.9% for patients with high grade (70% to 99%) stenosis. Of 13 ipsilateral strokes occurring after 30 days, 3 resulted in death. Of these strokes, 76.9% (10 of 13) occurred within the first 6 months of the stenting procedure and no events were recorded after 12 months. An additional 9 patients experienced ipsilateral transient ischemic attack after 30 days. Most postprocedural events (86%) could be attributed to interruption of antiplatelet medications (n=6), in-stent restenosis (n=12), or both (n=1). In 3 patients, the events were of uncertain etiology. CONCLUSIONS: After successful Wingspan percutaneous transluminal angioplasty and stenting, some patients continued to experience ipsilateral ischemic events. Most of these ischemic events occurred within 6 months of the procedure and were associated with the interruption of antiplatelet therapy or in-stent restenosis. PMID- 21636813 TI - Recombinant tissue-type plasminogen activator use for ischemic stroke in the United States: a doubling of treatment rates over the course of 5 years. AB - BACKGROUND AND PURPOSE: Recombinant tissue-type plasminogen activator (rtPA) is the only approved therapy for acute ischemic stroke (AIS). In 2004, 1.8% to 2.1% of AIS patients in the United States received rtPA. Given incentives from regulatory agencies and payors to increase rtPA use, we hypothesized that rtPA use in the United States would increase from 2005 to 2009. METHODS: AIS cases were defined by exclusion of hemorrhagic stroke and transient ischemic attack International Classification of Diseases 9th revision codes (430, 431, 432, and 435) from diagnosis-related groups 14, 15, 524, and 559 discharges. Patients receiving thrombolytics were identified using International Classification of Diseases 9th revision code 99.10 (Medicare Provider and Analysis Review and Premier databases) and pharmacy billing records (Premier). Change over time and differences between databases were tested using negative binomial regression. RESULTS: Within Medicare Provider and Analysis Review, thrombolytic use increased from 1.1% in 2005 to 3.4% in 2009 (P<0.001 for trend). Within Premier, thrombolytic use increased from 1.4% in 2005 to 3.7% in 2009 for all cases (P<0.001). Analysis of pharmacy billing records in Premier for 50-mg or 100-mg doses of rtPA showed that 3.4% of AIS cases were treated in 2009. Inclusion of patients with transient ischemic attack or hemorrhagic stroke International Classification of Diseases 9th revision codes who received any thrombolytic as "ischemic stroke patients receiving rtPA" changed the rate of thrombolysis to 5.2%. CONCLUSIONS: In 2009, 3.4% to 5.2% of AIS patients in the United States received thrombolytics, approximately double the rate of treatment in 2005. Rapid recognition and transport and quick treatment in the emergency department remain goals for further improving treatment rates. PMID- 21636814 TI - Outcome of patients with >=70% symptomatic intracranial stenosis after Wingspan stenting. AB - BACKGROUND AND PURPOSE: There were limited data on the long-term outcome of patients with symptomatic intracranial atherosclerotic stenosis >=70% after Wingspan stenting. Using our Wingspan cohort data and the data from the Warfarin and Aspirin for Symptomatic Intracranial Atherosclerotic Disease (WASID) as a historical control, we tested the hypothesis that stenting provided no benefit over antithrombotic therapy alone for these high-risk patients. METHODS: Between January 2007 and February 2009, 100 consecutive patients with intracranial atherosclerotic stenosis >=70% and symptoms within 90 days were enrolled into this prospective single-center Wingspan cohort study and followed up until the end of February 2010. Stenosis was measured per the WASID criteria. One-year risk of primary end point (any stroke or death within 30 days and ipsilateral ischemic stroke afterward) was compared with that of ipsilateral ischemic stroke in the WASID patients with >=70% stenosis. RESULTS: The stent placement success rate was 99%. All patients but 1 had clinical follow-up of >=12 months. During a mean follow-up of 1.8 years, 9 patients developed primary end point events (5 within 30 days and 4 afterward). The 1-year risk of the outcome events was lower than that in similar WASID patients: 7.3% (95% CI, 2.0% to 12.5%) versus 18% (95% CI, 13% to 24%; P<0.05). CONCLUSIONS: The clinical outcome of Wingspan stenting for high-risk intracranial atherosclerotic stenosis patients in this high-volume center study compares favorably with that of antithrombotic therapy alone. A randomized trial comparing medical therapy alone with medical therapy plus Wingspan stenting, conducted at high-volume centers, is needed to confirm the stenting benefit. PMID- 21636815 TI - Reliability and validity of bilateral ankle accelerometer algorithms for activity recognition and walking speed after stroke. AB - BACKGROUND AND PURPOSE: Outcome measures of mobility for large stroke trials are limited to timed walks for short distances in a laboratory, step counters and ordinal scales of disability and quality of life. Continuous monitoring and outcome measurements of the type and quantity of activity in the community would provide direct data about daily performance, including compliance with exercise and skills practice during routine care and clinical trials. METHODS: Twelve adults with impaired ambulation from hemiparetic stroke and 6 healthy controls wore triaxial accelerometers on their ankles. Walking speed for repeated outdoor walks was determined by machine-learning algorithms and compared to a stopwatch calculation of speed for distances not known to the algorithm. The reliability of recognizing walking, exercise, and cycling by the algorithms was compared to activity logs. RESULTS: A high correlation was found between stopwatch-measured outdoor walking speed and algorithm-calculated speed (Pearson coefficient, 0.98; P=0.001) and for repeated measures of algorithm-derived walking speed (P=0.01). Bouts of walking >5 steps, variations in walking speed, cycling, stair climbing, and leg exercises were correctly identified during a day in the community. Compared to healthy subjects, those with stroke were, as expected, more sedentary and slower, and their gait revealed high paretic-to-unaffected leg swing ratios. CONCLUSIONS: Test-retest reliability and concurrent and construct validity are high for activity pattern-recognition Bayesian algorithms developed from inertial sensors. This ratio scale data can provide real-world monitoring and outcome measurements of lower extremity activities and walking speed for stroke and rehabilitation studies. PMID- 21636816 TI - Validation of in vivo magnetic resonance imaging blood-brain barrier permeability measurements by comparison with gold standard histology. AB - BACKGROUND AND PURPOSE: We sought to validate the blood-brain barrier permeability measurements extracted from perfusion-weighted MRI through a relatively simple and frequently applied model, the Patlak model, by comparison with gold standard histology in a rat model of ischemic stroke. METHODS: Eleven spontaneously hypertensive rats and 11 Wistar rats with unilateral 2-hour filament occlusion of the right middle cerebral artery underwent imaging during occlusion at 4 hours and 24 hours after reperfusion. Blood-brain barrier permeability was imaged by gradient echo imaging after the first pass of the contrast agent bolus and quantified by a Patlak analysis. Blood-brain barrier permeability was shown on histology by the extravasation of Evans blue on fluorescence microscopy sections matching location and orientation of MR images. Cresyl-violet staining was used to detect and characterize hemorrhage. Landmark based elastic image registration allowed a region-by-region comparison of permeability imaging at 24 hours with Evans blue extravasation and hemorrhage as detected on histological slides obtained immediately after the 24-hour image set. RESULTS: Permeability values in the nonischemic tissue (marginal mean +/- SE: 0.15 +/- 0.019 mL/min 100 g) were significantly lower compared to all permeability values in regions of Evans blue extravasation or hemorrhage. Permeability values in regions of weak Evans blue extravasation (0.23 +/- 0.016 mL/min 100 g) were significantly lower compared to permeability values of in regions of strong Evans blue extravasation (0.29 +/- 0.020 mL/min 100 g) and macroscopic hemorrhage (0.35 +/- 0.049 mL/min 100 g). Permeability values in regions of microscopic hemorrhage (0.26 +/- 0.024 mL/min 100 g) only differed significantly from values in regions of nonischemic tissue (0.15 +/- 0.019 mL/min 100 g). CONCLUSIONS: Areas of increased permeability measured in vivo by imaging coincide with blood-brain barrier disruption and hemorrhage observed on gold standard histology. PMID- 21636817 TI - A cost-utility analysis of mechanical thrombectomy as an adjunct to intravenous tissue-type plasminogen activator for acute large-vessel ischemic stroke. AB - BACKGROUND AND PURPOSE: Mechanical thrombectomy has the potential to improve recanalization rates and outcomes for patients with ischemic stroke, but potential gains could be offset by procedural complications and costs. We evaluated the cost and utility of combined intravenous (IV) tissue-type plasminogen activator (tPA) and mechanical thrombectomy compared to IV tPA alone for acute large-vessel ischemic stroke. METHODS: We constructed a decision tree for a hypothetical 68-year-old with a large-vessel ischemic stroke who is eligible for IV tPA. The interventional strategy was IV tPA, a cerebral angiogram, and mechanical thrombectomy and thrombolysis if indicated. Recanalization, hemorrhage complications, and outcomes for the interventional strategy were from the Multi-MERCI study. The medical strategy was IV tPA using inputs from a comprehensive systematic review. Costs were estimated from Medicare reimbursements. We modeled lifetime costs and utilities for disability using a Markov model and Monte-Carlo multivariable sensitivity analysis. RESULTS: For the baseline scenario, the recanalization rate was 72.9% for the interventional strategy and 46.2% for the medical strategy. For the interventional strategy, the symptomatic hemorrhage rate was 8.6% with recanalization and 15.4% without. For the medical strategy, the corresponding rates were 3.6% and 13.3%, respectively. The interventional strategy was cost-effective in 97.6% of simulations (incremental cost-effectiveness ratio $16 001/quality-adjusted life year; 95% CI, $2736-$39,232). CONCLUSIONS: Based on observational data, the combination of IV tPA and mechanical thrombectomy for large-vessel ischemic stroke appears to be cost-effective compared to IV tPA alone. These findings require additional validation with randomized trial data. PMID- 21636818 TI - Intracranial stenting with Wingspan: still awaiting a safe landing. PMID- 21636819 TI - Improved cerebral vasomotor reactivity after exercise training in hemiparetic stroke survivors. AB - BACKGROUND AND PURPOSE: Animal studies provide strong evidence that aerobic exercise training positively influences cerebral blood flow, but no human studies support the use of exercise for improving cerebral hemodynamics. This randomized study in stroke survivors assessed the effects of treadmill aerobic exercise training (TM) on cerebral blood flow parameters compared to a control intervention of nonaerobic stretching. METHODS: Thirty-eight participants (19 in TM group and 19 in control group) with remote stroke (>6 months) and mild to moderate gait deficits completed middle cerebral artery blood flow velocity measurements by transcranial Doppler ultrasonography before and after a 6-month intervention period. Middle cerebral artery blood flow velocity was assessed bilaterally during normocapnia and hypercapnia (6% CO2). Cerebral vasomotor reactivity (cVMR) was calculated as percent change in middle cerebral artery blood flow velocity from normocapnia to hypercapnia (cVMR percent) and as an index correcting percent change for absolute increase in end tidal CO2 (cVMR index). RESULTS: The TM group had significantly larger improvements than did controls for both ipsilesional and contralesional cVMR index (P<=0.05) and contralesional cVMR percent (P<=0.01). Statin users in the TM group (n=10) had higher baseline cVMR and lower training-induced cVMR change, indicating that cVMR change among those not using statins (n=9) primarily accounted for the between group effects. There was a 19% increase in Vo2 peak for the TM group compared to a 4% decrease in the control group (P<0.01), and peak fitness change correlated with cVMR change (r=0.55; P<0.05). CONCLUSIONS: Our data provide the first evidence to our knowledge of exercise-induced cVMR improvements in stroke survivors, implying a protective mechanism against recurrent stroke and other brain-related disorders. Statin use appears to regulate cVMR and the cVMR training response. PMID- 21636820 TI - Short-term anomia training and electrical brain stimulation. AB - BACKGROUND AND PURPOSE: Language training success in chronic aphasia remains only moderate. Electric brain stimulation may be a viable way to enhance treatment efficacy. METHODS: In a randomized, double-blind, sham-controlled crossover trial, we assessed if anodal transcranial direct current stimulation compared to cathodal transcranial direct current stimulation and sham stimulation over the right temporo-parietal cortex would improve the success of short-term high frequency anomia training. Twelve chronic poststroke aphasia patients were studied. Naming outcome was assessed after training and 2 weeks later. RESULTS: All training conditions led to a significant increase in naming ability, which was retained for at least 2 weeks after the end of the training. Application of anodal transcranial direct current stimulation significantly enhanced the overall training effect compared to sham stimulation. Baseline naming ability significantly predicted anodal transcranial direct current stimulation effects. CONCLUSIONS: Anodal transcranial direct current stimulation applied over the nonlanguage dominant hemisphere can enhance language training outcome in chronic aphasia. Clinical Trial Registration- URL: www.clinicaltrials.gov/. Unique identifier: NCT00822068. PMID- 21636821 TI - Impact of white matter lesions on physical functioning and fall risk in older people: a systematic review. AB - BACKGROUND AND PURPOSE: White matter lesions (WMLs) are common findings on neuroimaging in older people. This review systematically evaluates the published literature on the associations between WMLs and balance, gait, mobility, and falls in older people. METHODS: Studies were identified with searches of the MEDLINE databases. Articles reporting associations between WMLs and balance, gait, mobility, and falls in older people in cross-sectional and longitudinal studies were included. RESULTS: Thirty-one articles reporting data from 19 studies met the inclusion criteria. There were consistent findings from both cross-sectional and longitudinal studies indicating greater WML volumes are associated with impaired balance, slower gait, and reduced mobility. Most studies addressing regional WML distributions have reported that WMHs in the frontal lobe and periventricular regions show the strongest relationships with balance, gait, and mobility impairments. In relation to falls, a threshold effect was apparent in that only those with severe WML volumes were found to be at increased risk of falling. CONCLUSIONS: The findings of this systematic review indicate that WMLs are common and are significantly associated with impaired balance, gait, mobility, and falls in older people. In many studies, however, impaired mobility and increased fall risk are only evident in people who have the most severe degree of WMLs. PMID- 21636822 TI - Severity of intraventricular extension correlates with level of admission glucose after intracerebral hemorrhage. AB - BACKGROUND AND PURPOSE: Hyperglycemia after spontaneous intracerebral hemorrhage (ICH) is associated with poor outcome, but the pathophysiology of ICH-induced glucose dysregulation remains unclear. We sought to identify clinical and radiographic parameters of ICH that are associated with admission hyperglycemia. METHODS: Patients admitted to the Columbia University Medical Center Neurological Intensive Care Unit with spontaneous ICH between January 2009 and September 2010 were prospectively enrolled in the ICH Outcomes Project. Clinical, radiographic, and laboratory data were collected prospectively. Receiver operating characteristic analysis was used to identify the glucose level with optimal sensitivity and specificity for in-hospital mortality. Logistic and linear regression analyses were used to identify independent predictors of outcome measures where appropriate. RESULTS: One hundred four patients admitted during the study period were included in the analysis. Mean admission glucose level was 8.23 +/- 3.15 mmol/L (3.83 to 18.89 mmol/L) and 23.2% had a history of diabetes mellitus. Admission glucose was significantly associated with discharge (P=0.003) and 3-month mortality (P=0.002). Critical hyperglycemia defined at 10 mmol/L independently predicted discharge mortality (P=0.027; OR, 4.381; 95% CI, 1.186 to 16.174) and 3-month mortality (P=0.011; OR, 10.95; 95% CI, 1.886 to 62.41). Admission intraventricular extension score (P=0.038; OR, 1.117; 95% CI, 1.043 to 1.197) and diabetes mellitus (P=0.002; OR, 5.530; 95% CI, 1.833 to 16.689) were independent predictors of critical hyperglycemia. The intraventricular extension score (B=0.115, P=0.001) linearly correlated with admission glucose level (R=0.612, P=0.001) after adjusting for other clinical variables. CONCLUSIONS: Admission hyperglycemia after spontaneous ICH is associated with poor outcome and potentially related to the presence and severity of intraventricular extension. PMID- 21636823 TI - Letter by Munin et al regarding article, "Botulinum Toxin for the Upper Limb after Stroke (BoTULS) trial: effect on impairment, activity limitation, and pain". PMID- 21636824 TI - Evaluation of left ventricular enlargement as a marker of early disease in familial dilated cardiomyopathy. AB - BACKGROUND: Echocardiographic screening of families with dilated cardiomyopathy has identified a subgroup of asymptomatic relatives with left ventricular enlargement (LVE). The prognostic significance of LVE in this setting is incompletely understood. METHODS AND RESULTS: We evaluated 457 asymptomatic relatives in 128 dilated cardiomyopathy families and identified 110 individuals (24%) with LVE. Serial echocardiograms in 72 untreated LVE relatives showed that 9 individuals (13%) had development of dilated cardiomyopathy over 10 to 152 months (median, 52). Thirty LVE relatives and 30 age- and sex-matched healthy control subjects were evaluated using 2-dimensional and M-mode echocardiography, tissue Doppler imaging, noninvasive pressure-volume assessment, exercise stress echocardiography, and brain natriuretic peptide levels. LVE relatives showed mild defects of systolic and diastolic LV function, with normal filling pressures and exercise-induced increments in systolic contraction in most cases. LV dimensions and fractional shortening most effectively differentiated LVE relatives from control subjects, with other functional indices lacking additive discriminative value. In a receiver operating characteristics analysis, the area under the curve for LV end-diastolic diameter (% predicted) was 0.96 (P<0.001). LV end-diastolic diameter (% predicted) >116% or LV end-diastolic diameter (% predicted) 112% to 116%+fractional shortening <=29% had high sensitivity (100%) and specificity (93%) for LVE relatives and identified 8 of 9 progressors. CONCLUSIONS: LVE is a common finding in asymptomatic relatives in dilated cardiomyopathy families and can be a marker of preclinical cardiomyopathy. Assessment of LV size and contractile function is required for differentiating between pathological and physiological causes of LVE and may help to identify those at risk of disease progression. PMID- 21636825 TI - Left renal venographic findings in children with orthostatic proteinuria. PMID- 21636826 TI - Overall and cardiovascular mortality in Norwegian kidney donors compared to the background population. AB - BACKGROUND: There are concerns regarding potential long-term risks to the living kidney donor. Cardiovascular mortality has not been evaluated. The aim of this study was to assess overall and cardiovascular mortality in previous kidney donors compared with a general population sample. METHODS: All live kidney donors in Norway in the period 1963-2007 were included. Controls matched 3:1 for age, gender and year of birth were provided by Statistics, Norway. Cause of death was retrieved from the death master file. Vital status as of 1 January 2010 was provided for all participants, and cause of death was available until 1 January 2008. Comparative survival analyses were performed by Kaplan-Meier curves and log rank test. Age-stratified death rates were calculated and compared with a selected group with a health status hypothetically allowing donation. RESULTS: There were 2269 living kidney donors in the study period. At donation, mean age was 47.6 + 12.6 years, 41.3% were male. Median observation time was 14.3 years. A total of 324 donors died during the study period. Causes of death were similar for donors and controls. By Kaplan-Meier analysis, overall and cardiovascular mortality was lower for previous kidney donors than for matched controls (P < 0.001 and P = 0.004, respectively). Age-stratified death rates were elevated for the oldest group of donors. CONCLUSIONS: Overall and cardiovascular mortality results are partially reassuring. However, the seemingly elevated mortality rate among the oldest donors requires further study. PMID- 21636827 TI - Health-related quality of life of hematopoietic stem cell transplant childhood survivors: state of the science. AB - The notion of health-related quality of life (HRQoL) holds unique significance in the treatment of patients who have undergone hematopoietic stem cell transplantation (HSCT). Not only is transplant procedure inevitably associated with immediate and late medical effects along with high mortality and morbidity rates, but it can also significantly affect the HRQoL for the patient and family. This review of literature will assist advanced practice nurses and pediatric oncology nurses in distinguishing and targeting interventions for patients and families who are at high risk of encountering distress during and following HSCT. It provides information on the assessment of pre-HSCT variables to identify patient subgroups in need of more aggressive supportive care to improve HRQoL during transplant. Furthermore, it serves as a guideline for developing interventional strategies and the role of the advanced practice nurse and pediatric oncology nurse caring for the patient throughout and following transplant. PMID- 21636828 TI - Pneumocystis carinii pneumonia prophylaxis: current therapies and recommendations. AB - Pneumocystis pneumonia (PCP) can be life threatening for children receiving chemotherapy and immunosuppressive medication, including high-dose steroids. Although there are no current guidelines for prophylaxis in pediatric oncology patients, ongoing studies are evaluating the efficacy, side effects, ease of administration, and compliance of drugs used for PCP prophylaxis. Drugs currently being prescribed in practice include Bactrim, pentamidine, dapsone, and atovaquone. Bactrim remains superior for preventing PCP, but alternatives are being analyzed and investigated for those unable to tolerate Bactrim because of drug allergy or side effects. Educating patients and families about the importance of PCP prophylaxis and compliance should be a priority for all health care providers caring for children receiving immunosuppressive medications, including chemotherapy. PMID- 21636829 TI - Developing a scale to measure parental worry and their attitudes toward childhood cancer after successful completion of treatment: a pilot study. AB - The purpose of this study is to develop and evaluate the psychometric characteristics of a scale designed to measure parents' attitude toward childhood cancer after treatment has ended. In this study, the 2 theoretical frameworks (theory of attitude and theory of worry) were used as they related to the study. An attitude is an idea charged with negative or positive emotion, directed to a psychological object, such as cancer. A sample of 84 White, middle class, American parents (n = 49 mothers, n = 35 fathers) of 51 childhood cancer survivors, whose treatment ended between 1988 and 2005, was surveyed between November 2005 and February 2006. Two factors were extracted using principal component analysis with oblique rotation. Cronbach's alpha reliability for Factor 1 was .91 and for Factor 2 was .76. This study suggests that most parents of cancer survivors tend to perseverate, ruminating on the idea that their child's cancer will return; as a result, they remain in a heightened state of alertness and develop uncontrollable thoughts, or inconsolable worry, about the recurrence of the disease. PMID- 21636830 TI - Mismatch between investigator-determined and patient-reported independence after spinal cord injury: consequences for rehabilitation and trials. AB - OBJECTIVE: This study investigated the course and relationship between investigator-determined and patient-reported level of independence within the first year after spinal cord injury (SCI). The authors examined variables that contributed to these scores. METHODS: In this observational cohort study, 73 patients with traumatic SCI were evaluated at 1, 3, and 6 months (and 40 subjects at 1 to 12 months). The investigator-determined independence was quantified using the Spinal Cord Independence Measure (SCIM). The subjective, patient-reported independence was determined by asking how their general restrictions influenced everyday life activities. Several variables were used to explain these 2 scores. RESULTS: The SCIM score was higher than the patient-reported independence and improved significantly more over time (up to about 70/100 at 12 months), whereas the perceived independence remained below 50/100. The correlations between the 2 measures were at most moderate (r(s) <= 0.51), but in general somewhat higher for subjects with tetraplegia. Age and muscle strength predicted the SCIM score well. No variable predicted the patient-reported level of independence. CONCLUSIONS: Investigator-determined and patient-reported outcomes can differ considerably and evolve differently. A patient-reported outcome measure may not detect actual functional improvement. It is likely that changes in patient-reported outcomes are influenced by many factors in addition to those associated with functional recovery, including psychological factors. PMID- 21636831 TI - Comparison of motor control deficits during treadmill and overground walking poststroke. AB - BACKGROUND: Force-sensing split-belt treadmills (TMs) provide an alternative to the conventional overground (OG) setting and allow new avenues for analyzing the biomechanics and motor control of walking. However, walking control may differ on a TM compared with walking OG. OBJECTIVE: To compare spatiotemporal, kinematic, and EMG-based measures of motor control between TM and OG walking at self selected and fastest comfortable speeds in persons with poststroke hemiparesis. METHODS: Individuals with chronic hemiparesis (56) and similarly aged healthy individuals (17) walked over an instrumented walkway and on an instrumented split belt TM; 16 channels of EMG recorded bilateral muscle activity, and a 12-camera motion capture system collected bilateral 3D kinematics. The authors applied a nonnegative matrix factorization (NNMF) algorithm to examine the underlying patterns of motor control. RESULTS: Self-selected walking patterns differed on the TM versus OG in controls: speed decreased, stride length decreased, stance percentage increased, and double-support percentage increased. Poststroke, responses were similar, but cadence also decreased, and step length asymmetry increased. Kinematic patterns were similar except those associated with slower walking speeds. NNMF demonstrated similar EMG variance in the 2 environments. CONCLUSION: Persons, both healthy and poststroke, walk with different gait parameters on the TM. Although measures of motor control were mostly similar between the 2 environments, the TM induced step length asymmetry in 30% of participants (60% of whom took longer paretic steps). TM walking, therefore, is a valid method for detecting motor control deficits. PMID- 21636832 TI - Novel expression of CST1 as candidate senescence marker. AB - Senescent cells exhibit altered expression of numerous genes. Identifying the significance of the changes in gene expression may help advance our understanding of the senescence biology. Here, we report on the consistent and strong upregulation of CST1 expression during cellular senescence, independent of the initial trigger. CST1 expression at both the messenger RNA and protein levels was barely detected in control cells, which included early passage proliferating, quiescent, or immortal human fibroblasts and various human tumor cell lines. Immunoblotting and immunofluorescence cytochemical studies further suggest that CST1 accumulates intracellularly, within vesicular structures. We discuss these results in light of the known function of CST1 as a potent inhibitor of lysosomal cysteine proteases. PMID- 21636834 TI - Fertility and intergroup bias in racial and minimal-group contexts: evidence for shared architecture. AB - Recent research has shown that White women's bias against Black men increases with elevated fertility across the menstrual cycle. We demonstrate that the association between fertility and intergroup bias is not limited to groups defined by race, but extends to group categories that are minimally defined, and may depend on the extent to which women associate out-group men with physical formidability. In Study 1, Black and White women with strong associations between the racial out-group and physical formidability displayed greater bias against out-group men as conception risk increased. Study 2 replicated these results in a minimal-group paradigm. These findings are consistent with the notion that women may be endowed with a psychological system that generates intergroup bias via mechanisms that rely on categorization heuristics and perceptions of the physical formidability of out-group men, particularly when the costs of sexual coercion are high. PMID- 21636833 TI - Usefulness of preclinical models for assessing the efficacy of late-life interventions for sarcopenia. AB - Caloric restriction and physical exercise have proven beneficial against age associated changes in body composition and declining physical performance; however, little is known regarding what benefit these interventions might have when initiated late in life. The study of mimetics of diet and exercise and the combination thereof may provide additional treatments for a vulnerable elderly population; however, how and when to initiate such interventions requires consideration in developing the most safe and efficacious treatment strategies. In this review, we focus on preclinical late-life intervention studies, which assess the relationship between physical function, sarcopenia, and body composition. We provide a conceptual framework for the ever-changing definition of sarcopenia and a rationale for the use of an appropriate rodent model of this condition. We finish by providing our perspective regarding the implications of this body of work and future areas of research that may also contribute to the ultimate goal of extending healthspan. PMID- 21636835 TI - Differential regulation of CIDEA and CIDEC expression by insulin via Akt1/2- and JNK2-dependent pathways in human adipocytes. AB - Both insulin and the cell death-inducing DNA fragmentation factor-alpha-like effector (CIDE) family play important roles in apoptosis and lipid droplet formation. Previously, we reported that CIDEA and CIDEC are differentially regulated by insulin and contribute separately to insulin-induced anti-apoptosis and lipid droplet formation in human adipocytes. However, the upstream signals of CIDE proteins remain unclear. Here, we investigated the signaling molecules involved in insulin regulation of CIDEA and CIDEC expression. The phosphatidylinositol 3-kinase (PI3K) inhibitors wortmannin and PI-103 blocked both insulin-induced downregulation of CIDEA and upregulation of CIDEC. The Akt inhibitor API-2 and the c-Jun N-terminal kinase (JNK) inhibitor SP600125 selectively inhibited insulin regulation of CIDEA and CIDEC expression, respectively, whereas the MAPK/ERK kinase inhibitor U0126 and the p38 inhibitor SB203580 did not. Small interfering RNA-mediated depletion of Akt1/2 prevented insulin-induced downregulation of CIDEA and inhibition of apoptosis. Depletion of JNK2, but not JNK1, inhibited insulin-induced upregulation of CIDEC and lipid droplet enlargement. Furthermore, insulin increased both Akt and JNK phosphorylation, which was abrogated by the PI3K inhibitors. These results suggest that insulin regulates CIDEA and CIDEC expression via PI3K, and it regulates expression of each protein via Akt1/2- and JNK2-dependent pathways, respectively, in human adipocytes. PMID- 21636836 TI - Potential of an electric prosthesis for dynamic facial reanimation. AB - Chronic facial paralysis is a devastating condition with severe functional and emotional consequences. The current surgical armamentarium permits the predictable reestablishment of a protective blink as well as good resting symmetry. Yet the ultimate goal of symmetric, spontaneous emotional expression remains elusive despite significant progress in the areas of peripheral nerve grafting and free tissue transfer. This commentary explores the possibility of an implantable electrical prosthesis for facial reanimation. It reviews animal studies supporting this concept as well as recent human data suggesting that such an implant could rescue denervated facial musculature, thus overcoming a major hurdle for existing reanimation techniques. PMID- 21636837 TI - Nonexophytic nasopharyngeal carcinoma: high frequency of advanced lymph node and distant metastasis. AB - OBJECTIVE: Nasopharyngeal carcinoma (NPC) usually presents with an exophytic mass. However, some patients present with biopsy-proven NPC but normal-appearing nasopharynx on endoscopic examination (nonexophytic NPC). The authors aim to characterize the clinicopathological features and survival outcomes of nonexophytic NPC in a high-incidence area. STUDY DESIGN AND SETTING: A retrospective cohort analysis of 274 patients with NPC in a tertiary institution. SUBJECTS AND METHODS: On the basis of endoscopic findings, the patients were categorized into nonexophytic and exophytic NPC. Clinical, pathological, and survival outcomes between the 2 groups were compared. RESULTS: Nonexophytic NPC was present in 24 patients (8.8%). A greater proportion of patients with nonexophytic NPC presented with T1 tumors compared to patients with exophytic NPC (70.8% vs 35.2%, P = .004). Similarly, a higher proportion of patients with nonexophytic NPC compared with exophytic NPC presented with both N3 and M1 disease (37.5% vs 13.2%, P = .01 and 25.0% vs 6%, P = .005, respectively). The mean duration of symptoms of patients did not differ significantly. The 5-year disease-specific and disease-free survivals did not show any significant differences. CONCLUSION: Nonexophytic NPC appears to be associated with higher risks of both N3 and M1 disease at presentation. However, these patients have similar treatment outcomes compared to patients with exophytic NPC. Although nonexophytic NPC appears to be an aggressive variant, current treatment protocols are as effective as for the group with exophytic NPC. Awareness of the existence of nonexophytic NPC is important for adequate diagnostic workup of patients with this subset of NPC. PMID- 21636838 TI - Treatment of velopharyngeal insufficiency by pharyngeal and velar fat injections. AB - The aim of this prospective study was to evaluate the effectiveness of fat injections in the treatment of velopharyngeal insufficiency (VPI). The study involved 10 patients (6 adults aged 19-48 years and 4 children aged 5-13 years) with mild/moderate VPI who were injected with 3.5 to 8 mL of fat in the posterior, lateral pharyngeal walls and soft palate under general anaesthesia. A second fat-grafting procedure was performed in 2 patients to achieve further improvement. Nasoendoscopy revealed a reduction in the closure gap in all patients, and the perceptual evaluation demonstrated improved speech intelligibility and resonance and reduced nasal air leakage in all cases (P < .005). The aerodynamic assessment showed a significant reduction in nasal airflow during phonation (P < .05). Follow-up was 6 to 23 months. In conclusion, fat injections improved voice resonance and reduced nasal air escape in all treated cases and can be a promising alternative to major procedures, such as velopharyngoplasties, for the treatment of mild/moderate VPI. PMID- 21636839 TI - Common foreign body, unusual site. PMID- 21636840 TI - An unusual presentation of a bronchial foreign body. PMID- 21636841 TI - Consideration of genetic contributions to the risk for spasmodic dysphonia. AB - Spasmodic dysphonia, a form of the neurologic condition known as dystonia, results from involuntary spasms of the larynx, producing interruptions of speech and changes in voice quality. The pathogenesis of spasmodic dysphonia is not well understood. However, several genetic mutations have been identified that cause different forms of dystonia. In some individuals, these genetic mutations result in spasmodic dysphonia, either with no other signs of dystonia or as part of a broader dystonia phenotype. Thus, research in the growing field of dystonia genetics may help to inform our understanding of the pathogenesis of spasmodic dysphonia. PMID- 21636842 TI - Modeling the measurements of cochlear microcirculation and hearing function after loud noise. AB - OBJECTIVE: Recent findings support the crucial role of microcirculatory disturbance and ischemia for hearing impairment especially after noise-induced hearing loss (NIHL). The aim of this study was to establish an animal model for in vivo analysis of cochlear microcirculation and hearing function after a loud noise to allow precise measurements of both parameters in vivo. STUDY DESIGN: Randomized controlled trial. Setting. Animal study. Subjects and Methods. After assessment of normacusis (0 minutes) using evoked auditory brainstem responses (ABRs), noise (106-dB sound pressure level [SPL]) was applied to both ears in 6 guinea pigs for 30 minutes while unexposed animals served as controls. In vivo fluorescence microscopy of the stria vascularis capillaries was performed after surgical exposure of 1 cochlea. ABR measurements were derived from the contralateral ear. RESULTS: After noise exposure, red blood cell velocity was reduced significantly by 24.3% (120 minutes) and further decreased to 44.5% at the end of the observation (210 minutes) in contrast to stable control measurements. Vessel diameters were not affected in both groups. A gradual decrease of segmental blood flow became significant (38.1%) after 150 minutes compared with controls. Hearing thresholds shifted significantly from 20.0 +/- 5.5 dB SPL (0 minutes) to 32.5 +/- 4.2 dB SPL (60 minutes) only in animals exposed to loud noise. CONCLUSION: With regard to novel treatments targeting the stria vascularis in NIHL, this standardized model allows us to analyze in detail cochlear microcirculation and hearing function in vivo. PMID- 21636843 TI - Seborrheic keratosis, keratotic type, of the external auditory canal. PMID- 21636844 TI - Diagnostic value of bent-lever planes in detecting abnormality of the malleus incus complex. AB - OBJECTIVE: To retrospectively investigate the diagnostic value of the bent-lever planes in detecting abnormality of the malleus-incus complex. STUDY DESIGN: Diagnostic test assessment. SETTING: The study was conducted at the Shandong Medical Imaging Research Institute, Shandong University, Jinan, People's Republic of China. SUBJECTS: Eighty-five ears in 74 patients with surgically proved abnormality of the malleus-incus complex. MATERIALS AND METHODS: The bent-lever planes and direct axial images were reviewed by 2 radiologists. The radiologists assigned a value of abnormality or continuity to different parts of the malleus incus complex. Differences in categorical data were evaluated with the McNemar test. The sensitivity, specificity, Youden index, and interobserver agreement were calculated. RESULTS: There was no significant difference between bent-lever planes and direct axial images in identifying abnormality of the malleus-incus complex. The sensitivity ranged from 71.4% to 97.1%, and specificity ranged from 89.5% to 100.0%. The Youden index ranged from 0.609 to 0.971. The radiologists had almost perfect or substantial agreement in identifying the abnormality of different parts of the malleus-incus complex. CONCLUSION: The bent-lever planes do not add additional diagnostic value for experienced observers. PMID- 21636845 TI - Systolic blood pressure and incident heart failure in the elderly. The Cardiovascular Health Study and the Health, Ageing and Body Composition Study. AB - BACKGROUND: The exact form of the association between systolic blood pressure (SBP) and heart failure (HF) risk in the elderly remains incompletely defined, especially in individuals not receiving antihypertensive drugs. OBJECTIVE: To examine the association between SBP and HF risk in the elderly. DESIGN: Competing risks proportional hazards modelling of incident HF risk, using 10-year follow-up data from two NIH-sponsored cohort studies: the Cardiovascular Health Study (inception: 1989-90 and 1992-3) and the Health ABC Study (inception: 1997-8). SETTING: Community-based cohorts. PARTICIPANTS: 4408 participants (age, 72.8 (4.9) years; 53.1% women, 81.7% white; 18.3% black) without prevalent HF and not receiving antihypertensive drugs at baseline. MAIN OUTCOME MEASURES: Incident HF, defined as first adjudicated hospitalisation for HF. RESULTS: Over 10 years, 493 (11.2%) participants developed HF. Prehypertension (120-139 mm Hg), stage 1 (140 159 mm Hg), and stage 2 (>=160 mm Hg) hypertension were associated with escalating HF risk; HRs versus optimal SBP (<120 mm Hg) in competing-risks models controlling for clinical characteristics were 1.63 (95% CI 1.23 to 2.16; p=0.001), 2.21 (95% CI 1.65 to 2.96; p<0.001) and 2.60 (95% CI 1.85 to 3.64; p<0.001), respectively. Overall 255/493 (51.7%) HF events occurred in participants with SBP <140 mm Hg at baseline. Increasing SBP was associated with higher HF risk in women than in men; no race-SBP interaction was seen. In analyses with continuous SBP, HF risk had a continuous positive association with SBP to levels as low as 113 mm Hg in men and 112 mm Hg in women. CONCLUSIONS: There is a continuous positive association between SBP and HF risk in the elderly for levels of SBP as low as <115 mm Hg; over half of incident HF events occur in individuals with SBP <140 mm Hg. PMID- 21636846 TI - Remote ischaemic conditioning before exercise: are we there yet? PMID- 21636847 TI - Effectiveness of local cold application on skin burns and pain after transthoracic cardioversion. AB - BACKGROUND: Transthoracic cardioversion (TTC) is widely used in emergency departments and daily clinical practice. TTC may cause skin lesions on the application of apical and sternal paddle areas. The lesions are characterised by redness, erythema and blister(s), and can be defined as first degree burns locally causing pain and increased sensitivity. AIM: To evaluate the effectiveness of local cold application on reducing the incidence, severity and pain/sensitivity of skin burns in patients who underwent TTC. METHODS: The study was conducted in the intensive care unit of the cardiovascular surgery department. The patients were assigned to study (n=24) and control groups (n=24). Local cold application was performed for a 1 hour period on patients in the study group, whereas only clinical procedures were applied in the control group following TTC. Incidence and severity of burn was evaluated 2 h after TTC, and pain/sensitivity scores were evaluated at 2, 4 and 24 h after TTC. Results The incidence of burn was significantly lower in the study group (3/24) compared to the control group (21/24) (12.5% vs 83.3%, p<0.001). Pain/sensitivity scores were significantly lower in the study group compared to the control group (p<0.05). CONCLUSION: Local cold application following TTC is an effective means of reducing the incidence and severity of burns and pain/sensitivity. It is cost effective and can easily be applied by nurses in medical/surgical units and emergency departments. PMID- 21636848 TI - Single-channel electroencephalography of epileptic seizures in the out-of hospital setting: an observational study. AB - OBJECTIVES: To evaluate whether single-channel electroencephalography (EEG) recording can be conducted in the out-of-hospital setting and whether it can be used to record electrographic signs of convulsive epileptic seizures. METHODS: This prospective observational feasibility study included patients who presented with a recent or ongoing epileptic seizure during out-of-hospital emergency treatment. Bifrontal single-channel EEG recordings were conducted by ambulance physicians throughout the initial treatment. The data recorded were analysed for the quality of recording and the occurrence of ictal EEG patterns. RESULTS: There were 45 adult patients who had a recent or an ongoing epileptic seizure in the study group and 15 patients with no neurological disorders in the control group. The median percentage of time during which no artefacts were detected by the device was 88.0% in the study group and 96.0% in the control group. EEG recordings for 3 out of 45 (6.6%) patients were of poor quality and not evaluable. Spike/wave or polyspike patterns were found in 98% and 100% of patients in the study and control groups, respectively, whereas the occurrence of periodic epileptiform discharges and delta waves with spikes showed a sensitivity and specificity of 100% (10/10) for the presence of an ongoing epileptic seizure. CONCLUSIONS: Single-channel EEG can be performed outside the hospital and yields useful recordings in most patients with acceptable rates of artefact. The diagnosis of generalised convulsive epileptic seizures by offline analysis of out of-hospital EEG showed a high sensitivity and specificity when compared with the clinical diagnosis. PMID- 21636849 TI - The frontoparietal attention network of the human brain: action, saliency, and a priority map of the environment. AB - The dorsal convexity of the human frontal and parietal lobes forms a network that is crucially involved in the selection of sensory contents by attention. This network comprehends cortex along the intraparietal sulcus, the inferior parietal lobe, and dorsal premotor cortex, including the frontal eye field. These regions are richly interconnected with recurrent fibers passing through the superior longitudinal fasciculus. The posterior parietal cortex has several functional characteristics-such as feature-independent coding, enhancement of activity by attention, representation of task-related signals, and access to multiple reference frames-that point to a central role of this region in the computation of a feature- and modality-independent priority map of the environment. The priority map integrates feature information elaborated in sensory cortex and top down representations of behavioral goals and expectations originating in the dorsolateral prefrontal and premotor cortex. This review presents converging evidence from single-unit studies of the primate brain, functional neuroimaging, and investigations of neuropsychological disorders such as Balint syndrome and spatial neglect for a decisive role of the frontoparietal attention network in the selection of relevant environmental information. PMID- 21636850 TI - Plasticity of cortical maps: multiple triggers for adaptive reorganization following brain damage and spinal cord injury. AB - Sensory and motor representations embedded in topographic cortical maps are use dependent, dynamically maintained, and self-organizing functional mosaics that constitute idiosyncratic entities involved in perceptual and motor learning abilities. Studies of cortical map plasticity have substantiated the view that local reorganization of sensory and motor areas has great significance in recovery of function following brain damage or spinal cord injury. In addition, the transfer of function to distributed cortical areas and subcortical structures represents an adaptive strategy for functional compensation. There is a growing consensus that subject-environment interactions, by continuously refining the canvas of synaptic connectivity and reshaping the anatomical and functional architecture of neural circuits, promote adaptive behavior throughout life. Taking advantage of use-dependent neural plasticity, early initiated rehabilitative procedures improve the potential for recovery. PMID- 21636851 TI - Epigenetic regulation of phosphatidylinositol 3,4,5-triphosphate-dependent Rac exchanger 1 gene expression in prostate cancer cells. AB - Aberrant up-regulation of P-Rex1 expression plays important roles in cancer progression and metastasis. The present study investigated the regulatory mechanism underlying P-Rex1 gene expression in prostate cancer cells. We showed that P-Rex1 expression was much higher in metastatic prostate cancer cells than in prostate epithelial cells and non-metastatic prostate cancer cells. Histone deacetylase (HDAC) inhibitors or silence of endogenous HDAC1 and HDAC2 markedly elevated P-Rex1 transcription in non-metastatic prostate cancer cells, whereas overexpression of recombinant HDAC1 in metastatic prostate cancer cells suppressed P-Rex1 expression. HDAC inhibitor trichostatin A (TSA) also significantly increased P-Rex1 promoter activity and caused acetylated histones to accumulate and associate with the P-Rex1 promoter. One Sp1 site, essential for basal promoter activity, was identified as critical for the TSA effect. TSA treatment did not alter the DNA-binding activity of Sp1 toward the P-Rex1 promoter; however, it facilitated the dissociation of the repressive HDAC1 and HDAC2 from the Sp1 binding region. Interestingly, HDAC1 association with Sp1 and with the P-Rex1 promoter were much weaker in metastatic prostate cancer PC-3 cells than in non-metastatic prostate cancer cells, and HDAC inhibitors only had very modest stimulatory effects on P-Rex1 promoter activity and P-Rex1 expression in PC-3 cells. Altogether, our studies demonstrate that HDACs could regulate P Rex1 gene transcription by interaction with Sp1 and by region-specific changes in histone acetylation within the P-Rex1 promoter. Disassociation of HDACs from Sp1 on the P-Rex1 promoter may contribute to aberrant up-regulation of P-Rex1 in cancer. PMID- 21636852 TI - NDRG4 protein-deficient mice exhibit spatial learning deficits and vulnerabilities to cerebral ischemia. AB - The N-myc downstream-regulated gene (NDRG) family consists of four related proteins, NDRG1-NDRG4, in mammals. We previously generated NDRG1-deficient mice that were unable to maintain myelin sheaths in peripheral nerves. This condition was consistent with human hereditary motor and sensory neuropathy, Charcot-Marie Tooth disease type 4D, caused by a nonsense mutation of NDRG1. In contrast, the effects of genetic defects of the other NDRG members remain unknown. In this study, we focused on NDRG4, which is specifically expressed in the brain and heart. In situ mRNA hybridization on the brain revealed that NDRG4 was expressed in neurons of various areas. We generated NDRG4-deficient mice that were born normally with the expected Mendelian frequency. Immunochemical analysis demonstrated that the cortex of the NDRG4-deficient mice contained decreased levels of brain-derived neurotrophic factor (BDNF) and normal levels of glial cell line-derived neurotrophic factor, NGF, neurotrophin-3, and TGF-beta1. Consistent with BDNF reduction, NDRG4-deficient mice had impaired spatial learning and memory but normal motor function in the Morris water maze test. When temporary focal ischemia of the brain was induced, the sizes of the infarct lesions were larger, and the neurological deficits were more severe in NDRG4 deficient mice compared with the control mice. These findings indicate that NDRG4 contributes to the maintenance of intracerebral BDNF levels within the normal range, which is necessary for the preservation of spatial learning and the resistance to neuronal cell death caused by ischemic stress. PMID- 21636853 TI - Role of the nuclear receptor coactivator AIB1-Delta4 splice variant in the control of gene transcription. AB - The oncogene amplified in breast cancer 1 (AIB1) is a nuclear receptor coactivator that plays a major role in the progression of various cancers. We previously identified a splice variant of AIB1 called AIB1-Delta4 that is overexpressed in breast cancer. Using mass spectrometry, we define the translation initiation of AIB1-Delta4 at Met(224) of the full-length AIB1 sequence and have raised an antibody to a peptide representing the acetylated N terminus. We show that AIB1-Delta4 is predominantly localized in the cytoplasm, although leptomycin B nuclear export inhibition demonstrates that AIB1-Delta4 can enter and traffic through the nucleus. Our data indicate an import mechanism enhanced by other coactivators such as p300/CBP. We report that the endogenously and exogenously expressed AIB1-Delta4 is recruited as efficiently as full-length AIB1 to estrogen-response elements of genes, and it enhances estrogen-dependent transcription more effectively than AIB1. Expression of an N-terminal AIB1 protein fragment, which is lost in the AIB1-Delta4 isoform, potentiates AIB1 as a coactivator. This suggests a model whereby the transcriptional activity of AIB1 is squelched by a repressive mechanism utilizing the N-terminal domain and that the increased coactivator function of AIB1-Delta4 is due to the loss of this inhibitory domain. Finally, we show, using Scorpion primer technology, that AIB1 Delta4 expression is correlated with metastatic capability of human cancer cell lines. PMID- 21636854 TI - Three-dimensional structure of the signal peptide peptidase. AB - Signal peptide peptidase (SPP) is an atypical aspartic protease that hydrolyzes peptide bonds within the transmembrane domain of substrates and is implicated in several biological and pathological functions. Here, we analyzed the structure of human SPP by electron microscopy and reconstructed the three-dimensional structure at a resolution of 22 A. Enzymatically active SPP forms a slender, bullet-shaped homotetramer with dimensions of 85 * 85 * 130 A. The SPP complex has four concaves on the rhombus-like sides, connected to a large chamber inside the molecule. Intriguingly, the N-terminal region of SPP is sufficient for the tetrameric assembly. Moreover, overexpression of the N-terminal region inhibited the formation of the endogenous SPP tetramer and the proteolytic activity within cells. These data suggest that the homotetramer is the functional unit of SPP and that its N-terminal region, which works as the structural scaffold, has a novel modulatory function for the intramembrane-cleaving activity of SPP. PMID- 21636855 TI - p16INK4a deficiency promotes IL-4-induced polarization and inhibits proinflammatory signaling in macrophages. AB - The CDKN2A locus, which contains the tumor suppressor gene p16(INK4a), is associated with an increased risk of age-related inflammatory diseases, such as cardiovascular disease and type 2 diabetes, in which macrophages play a crucial role. Monocytes can polarize toward classically (CAMphi) or alternatively (AAMphi) activated macrophages. However, the molecular mechanisms underlying the acquisition of these phenotypes are not well defined. Here, we show that p16(INK4a) deficiency (p16(-/-)) modulates the macrophage phenotype. Transcriptome analysis revealed that p16(-/-) BM-derived macrophages (BMDMs) exhibit a phenotype resembling IL-4-induced macrophage polarization. In line with this observation, p16(-/-) BMDMs displayed a decreased response to classically polarizing IFNgamma and LPS and an increased sensitivity to alternative polarization by IL-4. Furthermore, mice transplanted with p16(-/-) BM displayed higher hepatic AAMphi marker expression levels on Schistosoma mansoni infection, an in vivo model of AAMphi phenotype skewing. Surprisingly, p16(-/-) BMDMs did not display increased IL-4-induced STAT6 signaling, but decreased IFNgamma induced STAT1 and lipopolysaccharide (LPS)-induced IKKalpha,beta phosphorylation. This decrease correlated with decreased JAK2 phosphorylation and with higher levels of inhibitory acetylation of STAT1 and IKKalpha,beta. These findings identify p16(INK4a) as a modulator of macrophage activation and polarization via the JAK2-STAT1 pathway with possible roles in inflammatory diseases. PMID- 21636856 TI - Steroid-refractory GVHD: T-cell attack within a vulnerable endothelial system. AB - Acute graft-versus-host disease (GVHD) is a major complication of allogeneic stem cell transplantation (SCT) and can be readily controlled by systemic high-dose steroids in many patients. However, patients whose GVHD is refractory to this therapy have a poor prognosis. Refractory patients have ongoing end-organ damage despite effective immunosuppression with second-line regimens, suggesting pathomechanisms independent from the initiating T-cell attack. To explore whether endothelial damage might contribute to GVHD refractoriness and to study the role of angiopoietin-2 (ANG2) in this process, we have compared kinetics of T-cell activation markers and markers of endothelial dysfunction in the serum of patients with sensitive (n = 23) and refractory GVHD (n = 25). Longitudinal measurements of soluble FAS ligand along with other immune markers demonstrate that refractory patients are not exposed to an overwhelming or unresponsive T cell attack. However, in contrast to sensitive GVHD, refractory GVHD was associated with rising thrombomodulin levels and high ANG2/ vascular endothelial derived growth factor ratios. Patients with refractory GVHD showed significantly increased ANG2 levels already before SCT. These results suggest that endothelial cell vulnerability and dysfunction, rather than refractory T-cell activity, drives treatment refractoriness of GVHD and opens new avenues for prediction and control of this devastating condition. PMID- 21636857 TI - SUMO conjugation of STAT1 protects cells from hyperresponsiveness to IFNgamma. AB - The biologic effects of IFNgamma are mediated by the transcription factor STAT1. The activity of STAT1 is inhibited by small ubiquitin-like modifier (SUMO) conjugation. This occurs both directly through decreasing STAT1 tyrosine phosphorylation and indirectly by facilitating STAT1 dephosphorylation consequential to increased STAT1 solubility because of suppressed paracrystal assembly. However, the physiologic implications of SUMO conjugation have remained unclear. Here, we used fibroblasts and bone marrow-derived macrophages (BMMs) from knockin mice expressing SUMO-free STAT1 to explore the consequences of STAT1 sumoylation for IFNgamma signaling. Our experiments demonstrated buffer property of paracrystals for activated STAT1, such that SUMO-mediated paracrystal dispersal profoundly reduced phosphorylation of STAT1, which affected both the activating tyrosine 701 and the transcription-enhancing serine 727. Accordingly, the curtailed STAT1 activity in the nucleus caused by SUMO conjugation resulted in diminished transcription of IFNgamma-responsive genes; and increased the IFNgamma concentration more than 100-fold required to trigger lipopolysaccharide induced cytotoxicity in bone marrow-derived macrophages. These experiments identify SUMO conjugation of STAT1 as a mechanism to permanently attenuate the IFNgamma sensitivity of cells, which prevents hyperresponsiveness to this cytokine and its potentially self-destructive consequences. This sets the mode of SUMO-mediated inhibition apart from the other negative STAT regulators known to date. PMID- 21636858 TI - miR-181b is a biomarker of disease progression in chronic lymphocytic leukemia. AB - MicroRNAs play a crucial role in chronic lymphocytic leukemia. We investigated whether microRNAs can discriminate patients with a progressive disease from patients with a stable disease. We analyzed microRNA expression on leukemic cells isolated from 358 sequential samples of 114 patients with either stable or progressive disease. We found that during the course of the disease the expression values of miR-181b, the most dysregulated microRNA, decreased in samples of patients with a progressive (P < .001, training and validation sets) but not in samples of patients with a stable disease (P = .3, training set; P = .2, validation set) over time. A drop of >= 50% between sequential samples and/or a miR-181b value <= 0.005 at the starting time point were significant to differentiate progressive from stable disease (P = .004, training set; P < .001, validation set). These parameters were associated with high risk of requiring treatment (risk ratio, 5.8; 95% confidence interval, 2.5-14.9). We also observed that miR-181b targets Mcl-1 protein and that the decrease of its expression inversely correlated with increased protein levels of MCL1 and BCL2 target genes. We conclude that parameters defined on the basis of the miR-181b expression values specify disease progression in chronic lymphocytic leukemia and are associated with clinical outcome. PMID- 21636859 TI - Rap1 promotes VEGFR2 activation and angiogenesis by a mechanism involving integrin alphavbeta3. AB - Vascular endothelial growth factor (VEGF) acting through VEGF receptor 2 (VEGFR2) on endothelial cells (ECs) is a key regulator of angiogenesis, a process essential for wound healing and tumor metastasis. Rap1a and Rap1b, 2 highly homologous small G proteins, are both required for angiogenesis in vivo and for normal EC responses to VEGF. Here we sought to determine the mechanism through which Rap1 promotes VEGF-mediated angiogenesis. Using lineage-restricted Rap1 knockout mice we show that Rap1-deficiency in endothelium leads to defective angiogenesis in vivo, in a dose-dependent manner. Using ECs obtained from Rap1 deficient mice we demonstrate that Rap1b promotes VEGF-VEGFR2 kinase activation and regulates integrin activation. Importantly, the Rap1b-dependent VEGF-VEGFR2 activation is in part mediated via integrin alpha(v)beta(3). Furthermore, in an in vivo model of zebrafish angiogenesis, we demonstrate that Rap1b is essential for the sprouting of intersomitic vessels, a process known to be dependent on VEGF signaling. Using 2 distinct pharmacologic VEGFR2 inhibitors we show that Rap1b and VEGFR2 act additively to control angiogenesis in vivo. We conclude that Rap1b promotes VEGF-mediated angiogenesis by promoting VEGFR2 activation in ECs via integrin alpha(v)beta(3). These results provide a novel insight into the role of Rap1 in VEGF signaling in ECs. PMID- 21636860 TI - G-CSF receptor activation of the Src kinase Lyn is mediated by Gab2 recruitment of the Shp2 phosphatase. AB - Src activation involves the coordinated regulation of positive and negative tyrosine phosphorylation sites. The mechanism whereby receptor tyrosine kinases, cytokine receptors, and integrins activate Src is not known. Here, we demonstrate that granulocyte colony-stimulating factor (G-CSF) activates Lyn, the predominant Src kinase in myeloid cells, through Gab2-mediated recruitment of Shp2. After G CSF stimulation, Lyn dynamically associates with Gab2 in a spatiotemporal manner. The dephosphorylation of phospho-Lyn Tyr507 was abrogated in Shp2-deficient cells transfected with the G-CSF receptor but intact in cells expressing phosphatase defective Shp2. Auto-phosphorylation of Lyn Tyr396 was impaired in cells treated with Gab2 siRNA. The constitutively activated Shp2E76A directed the dephosphorylation of phospho-Lyn Tyr507 in vitro. Tyr507 did not undergo dephosphorylation in G-CSF-stimulated cells expressing a mutant Gab2 unable to bind Shp2. We propose that Gab2 forms a complex with Lyn and after G-CSF stimulation, Gab2 recruits Shp2, which dephosphorylates phospho-Lyn Tyr507, leading to Lyn activation. PMID- 21636861 TI - A phase 2 study of the safety and efficacy of rituximab with plasma exchange in acute acquired thrombotic thrombocytopenic purpura. AB - The safety and efficacy of weekly rituximab 375 mg/m(2) (*4), given within 3 days of acute TTP admission, with standard therapy (PEX and steroids) was evaluated. Clinical outcomes were compared to historical controls (n = 40) who had not received rituximab. Within the trial group, 15 of 40 required ICU admission and 15% of all cases with the highest troponin T levels on admission were ventilated. Before the second rituximab infusion, 68% of cases had a platelet count > 50 * 10(9)/L and 38% > 150 * 10(9)/L. Fewer PEX were required in whites compared to nonwhite in the rituximab group (mean 14 vs 21, P = .0095). Inpatient stay was reduced by 7 days in the non-ICU trial cases compared to historical controls (P = .04), especially in whites, with a mean reduction of 7 days (P = .05). Ten percent of trial cases relapsed, median, 27 months (17-31 months), compared to 57% in historical controls, median 18 months (3-60 months; P = .0011). There were no excess infections or serious adverse events with rituximab. In conclusion, rituximab appears a safe and effective therapy. Inpatient stay and relapse are significantly reduced in the rituximab cohort. Rituximab should be considered in conjunction with standard therapy on acute presentation of TTP. This study was registered at www.clinicaltrials.gov as NCT009-3713. PMID- 21636862 TI - Humans are equal: lessons to learn from the World Health Organization prescriptive growth standards. PMID- 21636863 TI - Implications of using World Health Organization growth reference (2007) for identifying growth problems in Hong Kong children aged 6 to 18 years. AB - OBJECTIVE: To evaluate the implications of replacing Hong Kong's 1993 growth references (HK1993) with the World Health Organization's 2007 references (WHO2007) for children aged 6 to 18 years. DESIGN: Cross-sectional study. SETTING: Thirty-six randomly selected primary and secondary schools in Hong Kong. PARTICIPANTS: A total of 14 842 children and adolescents aged 6 to 18 years in Hong Kong during 2005/06. MAIN OUTCOME MEASURES: Creation of age-specific z scores for height, weight, and body mass index relative to HK1993 and WHO2007 references. RESULTS: Use of WHO2007 instead of HK1993 could classify an additional 1.4% children aged 6 to 10 years and 2.8% children aged 11 to 18 years as having a short stature. Using WHO2007, respective proportions that could be classified as underweight and obese increased by 3.5% and 2.1% among children aged 6 to 10 years, and 5.5% and 1.6% among children aged 11 to 18 years. CONCLUSIONS: Use of WHO2007 could increase clinical workload and patient and parent anxiety by 'over-diagnosing' short stature and underweight. Although WHO2007 may have a role in international comparative research, retention of HK1993 would seem appropriate from a clinical perspective. PMID- 21636864 TI - Arthroscopic treatment of popliteal cyst. AB - OBJECTIVE: To review the results of arthroscopic treatment of popliteal cysts in our centre and analyse outcomes including complications. DESIGN: Retrospective study. SETTING: University teaching hospital, Hong Kong. PATIENTS: From July 2007 to July 2009, 11 patients with symptomatic popliteal cysts were treated arthroscopically. All of them had preoperative magnetic resonance imaging to confirm the diagnosis, identify the valvular opening, and the associated intra articular pathology. We used the Rauschning and Lindgren criteria for evaluation. RESULTS: Intra-articular pathology like cartilage degeneration and meniscus tear were commonly associated with popliteal cysts. All patients achieved symptomatic improvement after treatment and the recurrence rate was low. No major complications were encountered. We failed to identify (and correct) any valvular opening in one patient. CONCLUSION: From our experience, we conclude that arthroscopic treatment of popliteal cyst with correction of the valvular opening and treatment of associated intra-articular pathology is effective and safe. PMID- 21636865 TI - Association of molecular marker O(6)Methylguanine DNA methyltransferase and concomitant chemoradiotherapy with survival in Southern Chinese glioblastoma patients. AB - OBJECTIVES: (1) To compare the survival of concomitant chemotherapy and radiotherapy with radiotherapy alone in Chinese patients with primary glioblastoma. (2) To determine the methylation status of O(6)Methylguanine DNA methyltransferase in Chinese primary glioblastoma, and to assess the prognostic value of O(6)Methylguanine DNA methyltransferase methylation status in such patients. DESIGN: Retrospective correlative analysis. SETTING: University teaching hospital, Hong Kong. PATIENTS: Patients diagnosed with histologically proven primary glioblastoma in the period of March 2005 to June 2007 were recruited. Genomic DNA was isolated from formalin-fixed and paraffin-embedded sections of glioblastoma tissues. Methylation-specific polymerase chain reaction for O(6)Methylguanine DNA methyltransferase was performed. Patients' information at presentation was collected (age, performance status, steroid use, extent of resection, complications, radiotherapy data, use of chemotherapy). Primary outcome was measured by overall survival while secondary outcome was measured by progression-free survival. Overall and progression-free survivals were estimated by the Kaplan-Meier technique. Outcomes were assessed for groups with and without concomitant chemoradiotherapy and for groups with and without O(6)Methylguanine DNA methyltransferase methylation. RESULTS: A total of 35 glioblastoma patients were recruited; 27 were male and 8 female. Their mean age was 50 years. In all, 17 received concomitant chemoradiotherapy, and 18 received radiotherapy only. Their median overall survival was 12 (range, 7-17) months and the median progression-free survival was 5 (range, 3-6) months. In the radiotherapy alone group, the median progression-free survival and overall survival was 4 (range, 3 5) months and 6 (range, 2-10) months, respectively. In the concomitant radiochemotherapy group, the median progression-free survival and overall survival was 6 (range, 2-10) months and 13 (range, 8-18) months, respectively. Fifteen (43%) of the tumour samples showed methylation of O(6)Methylguanine DNA methyltransferase. There was a trend towards overall longer survival in the group with methylated tumours compared to those with unmethylated tumours; respective values for median survival (ranges) were 17 (13-21) versus 10 (6-14) months (P=0.105). CONCLUSIONS: Our single-centre results indicated that Chinese glioblastoma patients who had received concomitant chemoradiotherapy showed a trend towards longer overall survival compared to those receiving radiotherapy alone. Approximately 43% of our Chinese glioblastoma samples showed methylation of O(6)Methylguanine DNA methyltransferase. O(6)Methylguanine DNA methyltransferase methylation may be a significant prognostic factor in Chinese glioblastoma patients. PMID- 21636866 TI - A synopsis of current haemophilia care in Hong Kong. AB - OBJECTIVE: To provide a synopsis of current haemophilia care in Hong Kong. DESIGN: Retrospective survey. SETTING: All haematology units of the Hospital Authority in Hong Kong. PATIENTS: All patients with haemophilia A and haemophilia B. RESULTS: To date, there were 222 mild-to-severe haemophilia patients (192 type A, 30 type B) under regular public care in Hong Kong (43% were considered severe, 33% moderate, and 24% mild), which gave a crude prevalence of 6.8/100 000 male inhabitants. A total of 12.8 million units of Factor VIII and 3 million units of Factor IX were prescribed annually. This amounts to 1.83 units of FVIII per capita of the population, which is comparable to that of other developed countries. Leading causes of mortality were human immunodeficiency virus-related complications (10 cases) and cerebral bleeding (2 cases). The life expectancy of patients with severe haemophilia in Hong Kong is improving; currently the oldest patient is 60 years old. Such improved survival may be due to enhanced factor availability, prompt treatment of bleeding episodes at home, safer factor products, and better antiviral treatment. Primary prophylaxis is the accepted standard of care for severe and moderate cases, and "Factor First" has become hospital policy. However, 12 patients continue to present treatment challenges, due to the documented presence of factor inhibitors. In all, 28, 100, and 14 cases respectively were positive for human immunodeficiency virus, hepatitis C virus, and hepatitis B virus; the youngest patients with the corresponding infections being 28, 13, and 22 years old. Comprehensive care with dedicated physiotherapy, surgical support, and radionucleotide synovectomy may reduce morbidity further. CONCLUSION: A multidisciplinary approach can further improve the future care for haemophilia patients in Hong Kong. PMID- 21636867 TI - Risk of development of diabetes mellitus in Chinese women with persistently impaired glucose tolerance after gestational diabetes. AB - OBJECTIVES: To estimate the cumulative incidence for the development of diabetes mellitus in Chinese women with persistently impaired glucose tolerance after gestational diabetes, and evaluate putative risk factors. DESIGN: Historical cohort study. SETTING: A regional hospital in Hong Kong. PATIENTS: Women with postpartum impaired glucose tolerance (as confirmed by a 75-gram oral glucose tolerance test 6 weeks after delivery) seen between January 2000 and December 2006. RESULTS: After a mean follow-up period of 52 (standard deviation, 22; range, 12-106) months, 47 (20%) of the 238 women converted to diabetes mellitus. Concomitant postpartum impaired fasting plasma glucose levels increased the risk of future diabetes mellitus by 3.5-fold (95% confidence interval, 1.7-7.0; P=0.001) when compared to those with postpartum impaired glucose tolerance only. Based on multivariate analysis, only antepartum and postpartum fasting plasma glucose levels predicted future development of diabetes mellitus. At 1 year after delivery in 95/159 (60%) of the women, glucose tolerance regressed to normal, while in only 9/159 (6%) it progressed to diabetes mellitus. At this stage, 29% of those with impaired glucose regulation (impaired glucose tolerance, impaired fasting glucose or both) compared to 2% of those whose glucose tolerance reverted to normal developed diabetes mellitus upon subsequent follow-up (P<0.001). In all, 24/159 (15%) fulfilled the definition of metabolic syndrome and its presence was associated with 4.7-fold increased risk of future diabetes mellitus (95% confidence interval, 1.7-13.4; P=0.004). CONCLUSIONS: Women with persistent postpartum impaired glucose tolerance after gestational diabetes have a high risk of developing diabetes mellitus. However, a significant proportion of these women regress to normal glucose tolerance 1 year after delivery, and their risk of progression to diabetes mellitus is lower than those with persistent impaired glucose regulation. Therefore, women with a history of gestational diabetes, particularly those with persistent glucose intolerance 6 weeks and 1 year after delivery, should have regular surveillance for the development of diabetes mellitus. PMID- 21636868 TI - Learning needs in a medical curriculum in Hong Kong. AB - OBJECTIVE: To survey medical graduates from the University of Hong Kong on how well they perceived their learning needs had been fulfilled by the old (before 1997) and the new (after 1997) curricula. DESIGN: Retrospective questionnaire survey. SETTING: The University of Hong Kong, Hong Kong. PARTICIPANTS: Medical graduates from the University of Hong Kong who graduated between 1997 and 2006 were invited to complete a questionnaire online or in paper form; 1997-2001 graduates were trained under the old curriculum, and 2002-2006 graduates under the new curriculum. RESULTS: The response rate was 23%. The survey showed that the graduates of both curricula felt that research skills, population health, and ophthalmology were not emphasised enough in the medical programme. In addition, some graduates of the old curriculum mentioned interpersonal skills, ethics and professionalism, and language skills, which were pinpointed in the curriculum reform in 1997. Some graduates of the new curriculum mentioned anatomy, microbiology, and diagnostic radiology. Graduates of both the old and the new curricula perceived the same top five areas as being lacking in their respective curricula, in relationship to their clinical career and personal growth, namely: business administration, law, professional English, life coaching, and humanities. A small percentage of graduates also took courses in these areas after graduation. CONCLUSIONS: The survey showed that the curriculum reform in 1997 at the University of Hong Kong had correctly pinpointed some of the learning needs. The survey also identified educational needs in the existing curriculum that need to be dealt with in the forthcoming curriculum reform in 2012. PMID- 21636869 TI - The development of a Health Call Centre in Hong Kong: a study on the perceived needs of patients. AB - OBJECTIVES: To assess the preference for the development of a Health Call Centre and the perceived needs of patients in Hong Kong. DESIGN: Cross-sectional, questionnaire-based survey. SETTING: Two general out-patient clinics from health facilities in a geographical region in Hong Kong. PARTICIPANTS: Patients aged 18 years or above were recruited for the study, which was conducted between January and March 2009. MAIN OUTCOME MEASURES: Patients' perspectives of a Health Call Centre, perceived needs for services, perceived health status, and socio demographic status. RESULTS: A total of 403 participants completed the questionnaire with a response rate of 78%. A total of 342 (85%) supported the development of a Health Call Centre. Providing basic health and chronic disease information, current health conditions and treatment information, and caregiver support advice were cited as the top perceived needs on the development of a Health Call Centre. Adjusting for age, education, and individual monthly income, participants aged 31 to 64 years (odds ratio=4.37; 95% confidence interval, 1.92 9.99; P<0.001) and caregivers (odds ratio=3.41; 95% confidence interval, 1.21 9.59; P=0.020) were more likely to use the Health Call Centre. Presence of chronic illness had no significant correlation with the use of a Health Call Centre (odds ratio=1.43; 95% confidence interval, 0.69-3.00; P=0.340). CONCLUSIONS: This is the first study to assess the preferences for the development of a Health Call Centre and the perceived needs of patients in Hong Kong. The majority supported the development of a Health Call Centre. Services provided could meet the general needs of all patients and caregivers, and be accessible to old people. PMID- 21636870 TI - Quality of care of nurse-led and allied health personnel-led primary care clinics. AB - OBJECTIVES: To review the literature regarding quality of care of nurse-led and allied health personnel-led primary care clinics with specific attention to the quality indicators for fall prevention, continence care, pulmonary rehabilitation, mental health, pharmaceutical care, and wound care services. DATA SOURCES: Literature search from 1990 to 2010 including Ovid Medline, Cochrane Database, RAND (Research and Development) Corporation Health Database, the ACOVE (Assessing the Care of Vulnerable Elders) project and clinical guidelines from the United Kingdom, Australia, Canada, and the United States. STUDY SELECTION: This review was limited to studies involving adult, primary care patients. Where available, evidence from systematic reviews and meta-analyses were used to synthesise findings. DATA EXTRACTION: Combinations of the following terms (and related terms) were used to identify studies: primary care, clinic, allied health, nurse-led, fall prevention, continence care, incontinence, chronic obstructive pulmonary disorder, pulmonary disease, respiratory rehabilitation, mental health, mental wellbeing, depression, anxiety, wound care, leg ulcer, venous ulcer, dressings clinic, wound clinic, medication review, pharmacist-led, pharmaceutical care. DATA SYNTHESIS: A total of 21 international guidelines and 33 studies were selected for data synthesis. Despite a lack of consistent outcomes data, it is apparent that certain aspects of organisational structure and clinical care processes are important though not necessarily sufficient indicators of quality of care, because they themselves can influence care outcomes. Seven key factors were identified which seem important determinants of the quality of care provided by nurse- and allied health personnel-led clinics. CONCLUSION: Delivery of primary health care by nurse and allied health personnel led teams is a well-established model, internationally. Evidence from the literature provides benchmarks for standards of good practice. Knowledge of factors influencing quality of care can assist the planning, implementation, evaluation, and further expansion of such programmes, locally. PMID- 21636871 TI - Journey of a Hong Kong public teaching hospital in preparation of hospital accreditation. AB - Hospital accreditation is a new concept for Hong Kong Hospital Authority hospitals. Queen Mary Hospital has been engaged as one of the hospitals in a territory-wide Pilot Scheme of Hospital Accreditation. In preparation for accreditation, Queen Mary Hospital has undergone the process of self-assessment, staff engagement, and service improvements which all require well-planned strategies to achieve successful outcomes. In this article, we highlight the journey of preparation and the staff engagement exercise we conducted to attain full accreditation. We also highlight the obstacles, conundrums, and pitfalls we encountered, along with successful overcoming strategies and countermeasures we adopted, and quandaries to be avoided. Throughout the preparation, the hospital's senior executives insisted that achieving hospital accreditation was not the main focus, but rather an emphasis on how the Pilot Scheme would bring about organisational transformations in our culture, and thus foster quality, safety, effectiveness, and reliability of services. We hope our experience can provide a reference and be of value to other hospitals that will go through the journey in the future. PMID- 21636872 TI - Common association of haemolytic uraemic syndrome with invasive Streptococcus pneumoniae infection in five Chinese paediatric patients. AB - Haemolytic uraemic syndrome is an important cause of acute renal impairment in childhood. We review the incidence, and clinical and laboratory features of haemolytic uraemic syndrome in a Chinese population. Five patients were identified from 2006 to 2008. All patients were young children with associated invasive Streptococcus pneumoniae pulmonary infection. Serotypes 3, 14, and 19A were confirmed in four patients. The classical post-diarrhoeal form associated with Escherichia coli (O157:H7) infection was not seen. One patient died of acute respiratory failure. Streptococcus pneumoniae infection, as an associated condition in haemolytic uraemic syndrome, is important and relatively common in Chinese patients, especially among children. The acute clinical picture is similar to that reported in the western literature, except for an uncommon association with meningitis. The medium-term renal outcome of the Chinese population appears to be more favourable than the Caucasians. Widespread vaccination against Streptococcus pneumoniae may have resulted in changes in bacterial epidemiology and clinicians should be continuously aware of this severe disease. The use of washed blood components for transfusion in the acute stage requires further study. PMID- 21636873 TI - Giant myxoma causing right ventricular outflow tract obstruction. AB - Atrial cardiac myxoma is the most common benign cardiac tumour. Atrial myxoma most commonly arises from the left atrium and, less frequently, from the right atrium or both ventricles. Cardiac myxoma arising from the tricuspid valve is rare. These tumours can present with right heart failure as a result of right ventricular outflow tract obstruction. A high index of suspicion and appropriate investigations are necessary for making the correct diagnosis. Fatal complications such as embolisation and obstruction of the outflow tract and other intracardiac structures make prompt surgical intervention necessary. We report on a patient with a rare type of giant myxoma arising from the tricuspid valve. He underwent successful operation with en-bloc removal of the tumour, while preserving the integrity of the tricuspid valve. PMID- 21636874 TI - Hypertrophic pyloric stenosis in a newborn: a diagnostic dilemma. AB - Infants with hypertrophic pyloric stenosis typically present at 2 to 4 weeks of age with nonbilious projectile vomiting. Hypertrophic pyloric stenosis is exceedingly rare in newborn infants and is scarcely reported in literature. Also, the diagnostic criteria for ultrasonographic measurements in newborn infants have yet to be determined. This report is of a newborn infant with hypertrophic pyloric stenosis. The patient presented with high-volume non-bile-stained output from a nasogastric tube and a dilated gastric bubble on abdominal radiograph. Contrast study ruled out intestinal malrotation. Two ultrasound tests showed that the pyloric muscle thickness and pyloric canal length were within normal limits. Subsequent laparotomy showed a thickened pylorus and pyloromyotomy was performed. The patient showed marked improvement in feeding postoperatively. A high index of suspicion is required for newborn infants presenting with gastric outlet obstruction. Ultrasound and contrast studies provide additional information, but definitive diagnosis may only be available intra-operatively. PMID- 21636875 TI - A case of young-onset dementia. AB - Dementia is a major worldwide public health concern in view of the global ageing phenomenon. Dementia usually occurs in old age. However, if the symptoms occur in young patients, the diagnosis can be challenging. Posterior cortical atrophy is a variant of the Alzheimer's disease, which is described as a presenile disease affecting relatively late-middle-aged patients. A combination of clinical, neuropsychological, and neuroimaging techniques may facilitate making a diagnosis of this particular patient group, as demonstrated in this report. Although there is no effective disease-modifying agent for treating these patients to date, there may be considerable pressure to arrive at a quick and accurate diagnosis from the perspective of employment and insurance. PMID- 21636877 TI - The lesson of "BRIC". PMID- 21636876 TI - Hepatosplenic candidiasis complicating acute myeloid leukaemia. PMID- 21636878 TI - Proteotoxic stress targeted therapy (PSTT). PMID- 21636879 TI - Fabrication of 13-93 bioactive glass scaffolds for bone tissue engineering using indirect selective laser sintering. AB - Bioactive glasses are promising materials for bone scaffolds due to their ability to assist in tissue regeneration. When implanted in vivo, bioactive glasses can convert into hydroxyapatite, the main mineral constituent of human bone, and form a strong bond with the surrounding tissues, thus providing an advantage over polymer scaffold materials. Bone scaffold fabrication using additive manufacturing techniques can provide control over pore interconnectivity during fabrication of the scaffold, which helps in mimicking human trabecular bone. 13 93 glass, a third-generation bioactive material designed to accelerate the body's natural ability to heal itself, was used in the research described herein to fabricate bone scaffolds using the selective laser sintering (SLS) process. 13-93 glass mixed with stearic acid (as the polymer binder) by ball milling was used as the powder feedstock for the SLS machine. The fabricated green scaffolds underwent binder burnout to remove the stearic acid binder and were then sintered at temperatures between 675 degrees C and 695 degrees C. The sintered scaffolds had pore sizes ranging from 300 to 800 um with 50% apparent porosity and an average compressive strength of 20.4 MPa, which is excellent for non-load bearing applications and among the highest reported for an interconnected porous scaffold fabricated with bioactive glasses using the SLS process. The MTT labeling experiment and measurements of MTT formazan formation are evidence that the rough surface of SLS scaffolds provides a cell-friendly surface capable of supporting robust cell growth. PMID- 21636880 TI - Pulsed plasma polymerization for controlling shrinkage and surface composition of nanopores. AB - Solid-state nanopores have emerged as sensors for single molecules and these have been employed to examine the biophysical properties of an increasingly large variety of biomolecules. Herein we describe a novel and facile approach to precisely adjust the pore size, while simultaneously controlling the surface chemical composition of the solid-state nanopores. Specifically, nanopores fabricated using standard ion beam technology are shrunk to the requisite molecular dimensions via the deposition of highly conformal pulsed plasma generated thin polymeric films. The plasma treatment process provides accurate control of the pore size as the conformal film deposition depends linearly on the deposition time. Simultaneously, the pore and channel chemical compositions are controlled by appropriate selection of the gaseous monomer and plasma conditions employed in the deposition of the polymer films. The controlled pore shrinkage is characterized with high resolution AFM, and the film chemistry of the plasma generated polymers is analyzed with FTIR and XPS. The stability and practical utility of this new approach is demonstrated by successful single molecule sensing of double-stranded DNA. The process offers a viable new advance in the fabrication of tailored nanopores, in terms of both the pore size and surface composition, for usage in a wide range of emerging applications. PMID- 21636881 TI - Effects of electrons on the shape of nanopores prepared by focused electron beam induced etching. AB - The fabrication of nanometric pores with controlled size is important for applications such as single molecule detection. We have recently suggested the use of focused electron beam induced etching (FEBIE) for the preparation of such nanopores in silicon nitride membranes. The use of a scanning probe microscope as the electron beam source makes this technique comparably accessible, opening the way to widespread fabrication of nanopores. Since the shape of the nanopores is critically important for their performance, in this work we focus on its analysis and study the dependence of the nanopore shape on the electron beam acceleration voltage. We show that the nanopore adopts a funnel-like shape, with a central pore penetrating the entire membrane, surrounded by an extended shallow-etched region at the top of the membrane. While the internal nanopore size was found to depend on the electron acceleration voltage, the nanopore edges extended beyond the primary electron beam spot size due to long-range effects, such as radiolysis and diffusion. Moreover, the size of the peripheral-etched region was found to be less dependent on the acceleration voltage. We also found that chemical etching is the rate-limiting step of the process and is only slightly dependent on the acceleration voltage. Furthermore, due to the chemical etch process the chemical composition of the nanopore rims was found to maintain the bulk membrane composition. PMID- 21636882 TI - A nanocontact printing system for sub-100 nm aligned patterning. AB - Though many aspects of contact printing have been explored extensively since its invention, there are still hurdles to overcome for multilayer printing in the nanometer regime. Here we report on an aligned nanocontact printing (nCP) system that has demonstrated a sub-100 nm alignment capability by means of moire fringes and microspacers. To address issues in the stamp inking, we have devised a microfluidic apparatus based on the gradient capillary force for transport of ink solutions. The nCP system has been tested by printing nucleoside phosphoramidites on a nanopillar arrayed substrate. Although the nCP system was designed primarily for use in the fabrication of high density DNA nanoarrays, it has the potential to be applied to other fields of nanotechnology for nanoscale patterning. PMID- 21636883 TI - Extremely high response of electrostatically exfoliated few layer graphene to ammonia adsorption. AB - Extremely high gas sensing properties of p-type few layer graphene flakes exfoliated from highly oriented pyrolytic graphite have been demonstrated. The current response to ammonia adsorption is strongly dependent on film thickness and is higher than that for graphene by 1-8 orders of magnitude. A maximal response was found for sample thickness ~ 2 nm. The effect is attributed to the formation of multiple p-n-p junctions at the grain boundaries in the polycrystalline graphene flakes exposed to ammonia-containing ambient. PMID- 21636884 TI - Suspension and measurement of graphene and Bi2Se3 thin crystals. AB - Coupling high-quality suspended atomic membranes to specialized electrodes enables the investigation of many novel phenomena, such as spin or Cooper pair transport in these two-dimensional systems. However, many electrode materials are not stable in the acids that are used to dissolve underlying substrates. Here we present a versatile and powerful multilevel lithographical technique to suspend thin crystals, which can be applied to the vast majority of substrate, crystal and electrode materials. Using this technique, we fabricated suspended graphene devices with Al electrodes and a mobility of 5500 cm(2) V(-1) s(-1). We also demonstrate, for the first time, fabrication and measurement of a free-standing thin Bi(2)Se(3) crystal, which has low contact resistance to electrodes and a mobility of approximately > 580 cm(2) V(-1) s(-1). PMID- 21636885 TI - BMP-2 and ALP gene expression induced by a BMP-2 gene-fibronectin-apatite composite layer. AB - The bone morphogenetic protein 2 (BMP-2) gene delivery system with a gene fibronectin (Fn)-apatite composite layer was fabricated on the surface of a hydroxyapatite ceramic scaffold. The BMP-2 gene-Fn-apatite composite layer was coated on the scaffold using a supersaturated calcium phosphate solution supplemented with BMP-2 DNA and Fn. The scaffolds were ectopically implanted into the dorsal subcutaneous tissue of rats. Four weeks after the implantation, the hydroxyapatite scaffold coated with the BMP-2 gene-Fn-apatite composite layer showed improved gene expressions of BMP-2 and alkaline phosphatase as compared with the scaffold coated with the apatite layer. Although these results suggest the possibility of ectopic bone formation induced by the present gene delivery system, further study is necessary to prove this. PMID- 21636886 TI - In vitro mechanical integrity of hydroxyapatite coated magnesium alloy. AB - The mechanical integrity of resorbable implants during service, especially in load bearing orthopaedic applications, is critical. The high degradation rate of resorbable magnesium and magnesium-based implants in body fluid may potentially cause premature in-service failure. In this study, a magnesium alloy (AZ91) was potentiostatically coated with hydroxyapatite at different cathodic voltages in an attempt to enhance the mechanical integrity. The mechanical integrity of the uncoated and hydroxyapatite coated alloys was evaluated after in vitro testing of the coated samples in simulated body fluid (SBF). The uncoated alloy showed 40% loss in the mechanical strength after five days exposure to SBF. However, the hydroxyapatite coated alloy exposed to SBF showed 20% improvement in the mechanical strength as compared to that of the uncoated alloy. The alloy coated potentiostatically at -2 V performed better than the -3 V coated alloy. The cross sectional analysis of the coatings revealed relatively uniform coating thickness for the -2 V coated alloy, whereas the -3 V coated alloy exhibited areas of uneven coating. This can be attributed to the increase in hydrogen evolution on the alloy during -3 V coating as compared to -2 V coating. The scanning electron micrographs of the in vitro tested alloy revealed that hydroxyapatite coating significantly reduced the localized corrosion of the alloy, which is critical for better in-service mechanical integrity. Thus, the study suggests that the in vitro mechanical integrity of resorbable magnesium-based alloy can be improved by potentiostatic hydroxyapatite coating. PMID- 21636889 TI - Structure and biochemical characterization of proliferating cellular nuclear antigen from a parasitic protozoon. AB - Proliferating cellular nuclear antigen (PCNA) is a toroidal-shaped protein that is involved in cell-cycle control, DNA replication and DNA repair. Parasitic protozoa are early-diverged eukaryotes that are responsible for neglected diseases. In this work, a PCNA from a parasitic protozoon was identified, cloned and biochemically characterized and its crystal structure was determined. Structural and biochemical studies demonstrate that PCNA from Entamoeba histolytica assembles as a homotrimer that is able to interact with and stimulate the activity of a PCNA-interacting peptide-motif protein from E. histolytica, EhDNAligI. The data indicate a conservation of the biochemical mechanisms of PCNA mediated interactions between metazoa, yeast and parasitic protozoa. PMID- 21636890 TI - Scaffoldin-borne family 3b carbohydrate-binding module from the cellulosome of Bacteroides cellulosolvens: structural diversity and significance of calcium for carbohydrate binding. AB - The potent cellulose-binding modules of cellulosomal scaffoldin subunits belong to the greater family of carbohydrate-binding modules (CBMs). They have generally been classified as belonging to family 3a on the basis of sequence similarity. They form nine-stranded beta-sandwich structures with jelly-roll topology. The members of this family possess on their surface a planar array of aromatic amino acid residues (known as the linear strip) that form stacking interactions with the glucose rings of cellulose chains and have a conserved Ca(2+)-binding site. Intriguingly, the CBM3 from scaffoldin A (ScaA) of Bacteroides cellulosolvens exhibits alterations in sequence that make it more similar to the CBMs of free cellulolytic enzymes, which are classified into CBM family 3b. X-ray structural analysis was undertaken in order to examine the structural consequences of the sequence changes and the consequent family affiliation. The CBM3 crystallized in space group I4(1)22 with one molecule in the asymmetric unit, yielding diffraction to a resolution of 1.83 A using X-ray synchrotron radiation. Compared with the known structures of other scaffoldin-borne CBMs, a sequence insertion and deletion appear to compensate for each other as both contained an aromatic residue that is capable of contributing to cellulose binding; hence, even though there are alterations in the composition and localization of the aromatic residues in the linear strip its binding ability was not compromised. Interestingly, no Ca(2+) ions were detected in the conserved calcium-binding site, although the module was properly folded; this suggests that the structural role of Ca(2+) is less important than originally supposed. These observations indicate that despite their conserved function the scaffoldin-borne CBMs are more diverse in their sequences and structures than previously assumed. PMID- 21636891 TI - Mutation of the His ligand in mitoNEET stabilizes the 2Fe-2S cluster despite conformational heterogeneity in the ligand environment. AB - MitoNEET is the only identified Fe-S protein localized to the outer mitochondrial membrane and a 1.5 A resolution X-ray analysis has revealed a unique structure [Paddock et al. (2007), Proc. Natl Acad. Sci. USA, 104, 14342-14347]. The 2Fe-2S cluster is bound with a 3Cys-1His coordination which defines a new class of 2Fe 2S proteins. The hallmark feature of this class is the single noncysteine ligand His87, which when replaced by Cys decreases the redox potential (E(m)) by ~300 mV and increases the stability of the cluster by around sixfold. Unexpectedly, the pH dependence of the lifetime of the 2Fe-2S cluster remains the same as in the wild-type protein. Here, the crystal structure of H87C mitoNEET was determined to 1.7 A resolution (R factor = 18%) to investigate the structural basis of the changes in the properties of the 2Fe-2S cluster. In comparison to the wild type, structural changes are localized to the immediate vicinity of the cluster-binding region. Despite the increased stability, Cys87 displays two distinct conformations, with distances of 2.3 and 3.2 A between the S(gamma) and the outer Fe of the 2Fe-2S cluster. In addition, Lys55 exhibits multiple conformations in the H87C mutant protein. The structure and distinct characteristics of the H87C mutant provide a framework for further studies investigating the effects of mutation on the properties of the 2Fe-2S cluster in this new class of proteins. PMID- 21636892 TI - Contribution of the 80s loop of HIV-1 protease to the multidrug-resistance mechanism: crystallographic study of MDR769 HIV-1 protease variants. AB - The flexible flaps and the 80s loops (Pro79-Ile84) of HIV-1 protease are crucial in inhibitor binding. Previously, it was reported that the crystal structure of multidrug-resistant 769 (MDR769) HIV-1 protease shows a wide-open conformation of the flaps owing to conformational rigidity acquired by the accumulation of mutations. In the current study, the effect of mutations on the conformation of the 80s loop of MDR769 HIV-1 protease variants is reported. Alternate conformations of Pro81 (proline switch) with a root-mean-square deviation of 3 4.8 A in the C(alpha) atoms of the I10V mutant and a side chain with a 'flipped out' conformation in the A82F mutant cause distortion in the S1/S1' binding pockets that affects inhibitor binding. The A82S and A82T mutants show local changes in the electrostatics of inhibitor binding owing to the mutation from nonpolar to polar residues. In summary, the crystallographic studies of four variants of MDR769 HIV-1 protease presented in this article provide new insights towards understanding the drug-resistance mechanism as well as a basis for design of future protease inhibitors with enhanced potency. PMID- 21636893 TI - X-CHIP: an integrated platform for high-throughput protein crystallization and on the-chip X-ray diffraction data collection. AB - The X-CHIP (X-ray Crystallization High-throughput Integrated Platform) is a novel microchip that has been developed to combine multiple steps of the crystallographic pipeline from crystallization to diffraction data collection on a single device to streamline the entire process. The system has been designed for crystallization condition screening, visual crystal inspection, initial X-ray screening and data collection in a high-throughput fashion. X-ray diffraction data acquisition can be performed directly on-the-chip at room temperature using an in situ approach. The capabilities of the chip eliminate the necessity for manual crystal handling and cryoprotection of crystal samples, while allowing data collection from multiple crystals in the same drop. This technology would be especially beneficial for projects with large volumes of data, such as protein complex studies and fragment-based screening. The platform employs hydrophilic and hydrophobic concentric ring surfaces on a miniature plate transparent to visible light and X-rays to create a well defined and stable microbatch crystallization environment. The results of crystallization and data-collection experiments demonstrate that high-quality well diffracting crystals can be grown and high-resolution diffraction data sets can be collected using this technology. Furthermore, the quality of a single-wavelength anomalous dispersion data set collected with the X-CHIP at room temperature was sufficient to generate interpretable electron-density maps. This technology is highly resource-efficient owing to the use of nanolitre-scale drop volumes. It does not require any modification for most in-house and synchrotron beamline systems and offers a promising opportunity for full automation of the X-ray structure-determination process. PMID- 21636894 TI - Structural basis for drug and substrate specificity exhibited by FIV encoding a chimeric FIV/HIV protease. AB - A chimeric feline immunodeficiency virus (FIV) protease (PR) has been engineered that supports infectivity but confers sensitivity to the human immunodeficiency virus (HIV) PR inhibitors darunavir (DRV) and lopinavir (LPV). The 6s-98S PR has five replacements mimicking homologous residues in HIV PR and a sixth which mutated from Pro to Ser during selection. Crystal structures of the 6s-98S FIV PR chimera with DRV and LPV bound have been determined at 1.7 and 1.8 A resolution, respectively. The structures reveal the role of a flexible 90s loop and residue 98 in supporting Gag processing and infectivity and the roles of residue 37 in the active site and residues 55, 57 and 59 in the flap in conferring the ability to specifically recognize HIV PR drugs. Specifically, Ile37Val preserves tertiary structure but prevents steric clashes with DRV and LPV. Asn55Met and Val59Ile induce a distinct kink in the flap and a new hydrogen bond to DRV. Ile98Pro->Ser and Pro100Asn increase 90s loop flexibility, Gln99Val contributes hydrophobic contacts to DRV and LPV, and Pro100Asn forms compensatory hydrogen bonds. The chimeric PR exhibits a comparable number of hydrogen bonds, electrostatic interactions and hydrophobic contacts with DRV and LPV as in the corresponding HIV PR complexes, consistent with IC(50) values in the nanomolar range. PMID- 21636895 TI - Structural characterization of the PPIase domain of FKBP51, a cochaperone of human Hsp90. AB - Steroid hormone receptors are key components of mammalian stress and sex hormone systems. Many of them rely on the Hsp90 chaperone system for full function and are further fine-tuned by Hsp90-associated peptidyl-prolyl isomerases such as FK506-binding proteins 51 and 52. FK506-binding protein 51 (FKBP51) has been shown to reduce glucocorticoid receptor signalling and has been genetically associated with human stress resilience and with numerous psychiatric disorders. The peptidyl-prolyl isomerase domain of FKBP51 contains a high-affinity binding site for the natural products FK506 and rapamycin and has further been shown to convey most of the inhibitory activity on the glucocorticoid receptor. FKBP51 has therefore become a prime new target for the treatment of stress-related affective disorders that could be amenable to structure-based drug design. Here, a series of high-resolution structures of the peptidyl-prolyl isomerase domain of FKBP51 as well as a cocrystal structure with the prototypic ligand FK506 are described. These structures provide a detailed picture of the drug-binding domain of FKBP51 and the molecular binding mode of its ligand as a starting point for the rational design of improved inhibitors. PMID- 21636896 TI - Structure-based design of a disulfide-linked oligomeric form of the simian virus 40 (SV40) large T antigen DNA-binding domain. AB - The modular multifunctional protein large T antigen (T-ag) from simian virus 40 orchestrates many of the events needed for replication of the viral double stranded DNA genome. This protein assembles into single and double hexamers on specific DNA sequences located at the origin of replication. This complicated process begins when the origin-binding domain of large T antigen (T-ag ODB) binds the GAGGC sequences in the central region (site II) of the viral origin of replication. While many of the functions of purified T-ag OBD can be studied in isolation, it is primarily monomeric in solution and cannot assemble into hexamers. To overcome this limitation, the possibility of engineering intermolecular disulfide bonds in the origin-binding domain which could oligomerize in solution was investigated. A recent crystal structure of the wild type T-ag OBD showed that this domain forms a left-handed spiral in the crystal with six subunits per turn. Therefore, we analyzed the protein interface of this structure and identified two residues that could potentially support an intermolecular disulfide bond if changed to cysteines. SDS-PAGE analysis established that the mutant T-ag OBD formed higher oligomeric products in a redox dependent manner. In addition, the 1.7 A resolution crystal structure of the engineered disulfide-linked T-ag OBD is reported, which establishes that oligomerization took place in the expected manner. PMID- 21636897 TI - Analysis of phases in the structure determination of an icosahedral virus. AB - The constraints imposed on structure-factor phases by noncrystallographic symmetry (NCS) allow phase improvement, phase extension to higher resolution and hence ab initio phase determination. The more numerous the NCS redundancy and the greater the volume used for solvent flattening, the greater the power for phase determination. In a case analyzed here the icosahedral NCS phasing appeared to have broken down, although later successful phase extension was possible when the envelope around the NCS region was tightened. The phases from the failed phase determination attempt fell into four classes, all of which satisfied the NCS constraints. These four classes corresponded to the correct solution, opposite enantiomorph, Babinet inversion and opposite enantiomorph with Babinet inversion. These incorrect solutions can be seeded from structure factors belonging to reciprocal-space volumes that lie close to icosahedral NCS axes where the structure amplitudes tend to be large and the phases tend to be 0 or pi. Furthermore, the false solutions can spread more easily if there are large errors in defining the envelope designating the region in which NCS averaging is performed. PMID- 21636898 TI - Structures of human nucleosomes containing major histone H3 variants. AB - The nucleosome is the fundamental repeating unit of chromatin, via which genomic DNA is packaged into the nucleus in eukaryotes. In the nucleosome, two copies of each core histone, H2A, H2B, H3 and H4, form a histone octamer which wraps 146 base pairs of DNA around itself. All of the core histones except for histone H4 have nonallelic isoforms called histone variants. In humans, eight histone H3 variants, H3.1, H3.2, H3.3, H3T, H3.5, H3.X, H3.Y and CENP-A, have been reported to date. Previous studies have suggested that histone H3 variants possess distinct functions in the formation of specific chromosome regions and/or in the regulation of transcription and replication. H3.1, H3.2 and H3.3 are the most abundant H3 variants. Here, crystal structures of human nucleosomes containing either H3.2 or H3.3 have been solved. The structures were essentially the same as that of the H3.1 nucleosome. Since the amino-acid residues specific for H3.2 and H3.3 are located on the accessible surface of the H3/H4 tetramer, they may be potential interaction sites for H3.2- and H3.3-specific chaperones. PMID- 21636899 TI - High-resolution neutron crystallographic studies of the hydration of the coenzyme cob(II)alamin. AB - The hydration of the coenzyme cob(II)alamin has been studied using high resolution monochromatic neutron crystallographic data collected at room temperature to a resolution of 0.92 A on the original D19 diffractometer with a prototype 4 degrees * 64 degrees detector at the high-flux reactor neutron source run by the Institute Laue-Langevin. The resulting structure provides hydrogen-bonding parameters for the hydration of biomacromolecules to unprecedented accuracy. These experimental parameters will be used to define more accurate force fields for biomacromolecular structure refinement. The presence of a hydrophobic bowl motif surrounded by flexible side chains with terminal functional groups may be significant for the efficient scavenging of ligands. The feasibility of extending the resolution of this structure to ultrahigh resolution was investigated by collecting time-of-flight neutron crystallographic data during commissioning of the TOPAZ diffractometer with a prototype array of 14 modular 2 degrees * 21 degrees detectors at the Spallation Neutron Source run by Oak Ridge National Laboratory. PMID- 21636901 TI - Structure of an essential GTPase, YsxC, from Thermotoga maritima. AB - YsxC belongs to the YihA family of TRAFAC class GTPases. The protein is involved in the biogenesis of ribosomes and is essential for the survival of a wide range of bacteria. Here, crystal structures of YsxC from Thermotoga maritima and its complex with GDP were determined at maximal resolutions of 2.3 and 1.9 A, respectively. Major structural differences are observed in the switch I region, which is disordered in the apo form but exists in both an 'open' and a 'closed' conformation in the GDP-bound state. A comparison with the structure of the GMPPNP-YsxC complex from Bacillus subtilis provides insights into the mechanism of conformational change in the switch I and II regions upon hydrolysis of GTP. PMID- 21636902 TI - Structure of human R-state aquomethemoglobin at 2.0 A resolution. AB - The crystal structure of tetrameric (alphabeta)(2) R-state human adult aquomethemoglobin is reported at 2.0 A resolution. The asymmetric unit contained one alphabeta subunit pair. The R-state crystal belonged to space group P4(1)2(1)2, with unit-cell parameters a = b = 53.6, c = 192.8 A. An Fe-bound water molecule was modeled into the heme distal pockets of each of the alpha and beta subunits. In the alpha subunit, a highly ordered liganded water was modeled with an Fe-O(water) distance of 2.2 A and appears to be protected against escape from the distal pocket by the conformation of the heme propionate groups, which point upwards towards the distal His58 residue aided by a hydrogen-bonding network involving the solvent. In the beta subunit, the liganded water exhibited greater motion and was modeled with a longer Fe-O(water) distance of 2.5 A; in this subunit both propionate groups point downwards away from the distal His63 residue, presumably allowing greater motion of the liganded water in and out of the distal pocket. PMID- 21636903 TI - High-resolution structure of the recombinant sweet-tasting protein thaumatin I. AB - Thaumatin, an intensely sweet-tasting plant protein, elicits a sweet taste at a concentration of 50 nM. The crystal structure of a recombinant form of thaumatin I produced in the yeast Pichia pastoris has been determined to a resolution of 1.1 A. The model was refined with anisotropic B parameters and riding H atoms. A comparison of the diffraction data and refinement statistics for recombinant thaumatin I with those for plant thaumatin I revealed no significant differences in the diffraction data. The R values for recombinant thaumatin I and plant thaumatin I (F(o) > 4sigma) were 9.11% and 9.91%, respectively, indicating the final model to be of good quality. Notably, the electron-density maps around Asn46 and Ser63, which differ between thaumatin variants, were significantly improved. Furthermore, a number of H atoms became visible in an OMIT map and could be assigned. The high-quality structure of recombinant thaumatin with H atoms should provide details about sweetness determinants in thaumatin and provide valuable insights into the mechanism of its interaction with taste receptors. PMID- 21636904 TI - Preliminary X-ray crystallographic studies of the catalytic subunit of Escherichia coli AHAS II with its cofactors. AB - Acetohydroxyacid synthase (AHAS) is the first common enzyme in the branched-chain amino-acid biosynthesis pathway and is the target of several classes of commercial herbicides. In this study, the Escherichia coli ilvG gene that encodes the catalytic subunit of AHAS II was cloned into the pET28a vector and expressed in soluble form at high levels in E. coli strain BL21 (DE3) cells. The protein was purified using Ni(2+)-chelating chromatography followed by size-exclusion chromatography. The catalytic subunit of E. coli AHAS II was cocrystallized with its cofactors Mg(2+), FAD and ThDP using the sitting-drop vapour-diffusion method and the crystals diffracted to 2.80 A resolution. PMID- 21636905 TI - Crystallization and preliminary X-ray crystallographic analysis of Escherichia coli CheA P3 dimerization domain. AB - The chemotaxis histidine kinase CheA assembles into a dimer in which the P3 dimerization domain forms a four-helix bundle by the parallel association of two alpha-helical hairpins from each subunit. Ligand occupancy of the chemoreceptor regulates signal transduction by controlling the autophosphorylation activity of CheA. Autophosphorylation of CheA occurs in trans, i.e. one subunit phosphorylates the other. The P3 domain of CheA from Escherichia coli has been overexpressed in E. coli and crystallized at 298 K using PEG as a precipitant. X ray diffraction data to 2.80 A resolution have been collected at 100 K using synchrotron radiation. The crystal belonged to space group P1, with unit-cell parameters a = 59.271, b = 67.674, c = 82.815 A, alpha = 77.568, beta = 86.073, gamma = 64.436 degrees . The asymmetric unit may contain up to ten dimeric units of P3 four-helix bundles. PMID- 21636906 TI - Crystallization and preliminary X-ray analysis of geraniol dehydrogenase from Backhousia citriodora (lemon myrtle). AB - A recombinant form of geraniol dehydrogenase (EC 1.1.1.183) from Backhousia citriodora was overexpressed in Escherichia coli and purified and crystallized by the sitting-drop method using polyethylene glycol 3350 as a precipitant. A data set to 2.3 A resolution was collected from a monocrystal at 98 K using synchrotron radiation on beamline NE3A of the Photon Factory. The crystals belonged to the orthorhombic group P2(1)2(1)2, with unit-cell parameters a = 125.00, b = 151.01, c = 51.18 A. The asymmetric unit is expected to contain two BcGEDH molecules, with a corresponding crystal volume per protein weight of 3.1 A(3) Da(-1) and a solvent content of 60.6%. PMID- 21636907 TI - Expression, purification, crystallization and preliminary X-ray diffraction studies of phosphoglycerate kinase from methicillin-resistant Staphylococcus aureus MRSA252. AB - Phosphoglycerate kinase (PGK) from methicillin-resistant Staphylococcus aureus MRSA252 has been cloned in pQE30 expression vector, overexpressed in Escherichia coli SG13009 (pREP4) cells and purified to homogeneity. The protein was crystallized from 0.15 M CaCl(2), 0.1 M HEPES-NaOH pH 6.8, 20%(w/v) polyethylene glycol 2000 at 298 K by the hanging-drop vapour-diffusion method. The crystals belonged to space group P2(1), with unit-cell parameters a = 45.14, b = 74.75, c = 58.67 A, beta = 95.72 degrees . X-ray diffraction data have been collected and processed to a maximum resolution of 2.3 A. The presence of one molecule in the asymmetric unit gives a Matthews coefficient (V(M)) of 2.26 A(3) Da(-1) with a solvent content of 46%. The structure has been solved by molecular replacement and structure refinement is now in progress. PMID- 21636908 TI - Crystallization and preliminary X-ray analysis of Aspergillus oryzae catechol oxidase. AB - Catechol oxidase is an enzyme that catalyzes the oxidation of o-diphenols to the corresponding o-quinones. It is a copper-containing enzyme with a binuclear copper active site. Here, the crystallization and multiple-wavelength anomalous dispersion data collection of catechol oxidase from the mould fungus Aspergillus oryzae are described. During the purification, three forms of the enzyme (39.3, 40.5 and 44.3 kDa) were obtained. A mixture of these three forms was initially crystallized and gave crystals that diffracted to 2.5 A resolution and belonged to space group P3(2)21, with unit-cell parameters a = b = 118.9, c = 84.5 A, alpha = beta = 90, gamma = 120 degrees . A preparation containing only the shorter form (39.3 kDa) produced crystals that diffracted to 2.9 A resolution and belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 51.8, b = 95.3, c = 139.5 A, alpha = beta = gamma = 90 degrees . PMID- 21636909 TI - Crystallization and preliminary X-ray crystallographic analysis of Salmonella Typhimurium CueP. AB - Salmonella enterica serovar Typhimurium (S. Typhimurium) can survive in the phagosome of macrophages, causing serious medical and veterinary problems. CueP is uniquely found in S. Typhimurium and has been characterized as a major periplasmic copper-binding protein. Although cueP has been identified as being responsible for the copper resistance of the bacterium in vivo, the biochemical role and three-dimensional structure of CueP remain unknown. In this study, CueP from S. Typhimurium was overexpressed and the recombinant protein was purified using Ni-NTA affinity, anion-exchange and gel-filtration chromatographies. The purified CueP protein was crystallized using the vapour-diffusion method. A diffraction data set was collected to 2.5 A resolution at 100 K. The crystal belonged to space group P2(1)2(1)2(1). To obtain initial phases, selenomethionyl substituted protein was overproduced and purified. Optimization of crystallization conditions for the selenomethionyl-substituted protein is in progress. PMID- 21636910 TI - Crystallization of the factor H-binding protein, FhbB, from the periopathogen Treponema denticola. AB - Treponema denticola is a primary etiological agent of periodontal disease. T. denticola evades complement-mediated killing by binding to the host's factor H (FH), a negative regulator of the alternative complement pathway. The T. denticola FH-binding protein has been identified and designated as factor H binding protein B (FhbB). Crystals of recombinant FhbB were obtained by the hanging-drop vapor-diffusion method using sodium citrate and 0.2 M sodium thiocyanate. FhbB crystals diffracted to 1.8 A resolution and belonged to space group P4(3)2(1)2 or P4(1)2(1)2, with unit-cell parameters a = b = 46.76, c = 167.68 A. Two FhbB molecules per asymmetric unit gave a Matthews coefficient of 2.2 A(3) Da(-1) and a solvent content of 44%. FhbB is the smallest bacterially produced FH-binding protein identified to date. Determination of its structure will provide unique insight into the minimal structural determinants required for FH binding. PMID- 21636911 TI - Crystallization and preliminary X-ray analysis of argininosuccinate lyase from Streptococcus mutans. AB - Argininosuccinate lyase (ASL) is an important enzyme in arginine synthesis and the urea cycle, which are highly conserved from bacteria to eukaryotes. The gene encoding Streptococcus mutans ASL (smASL) was amplified and cloned into expression vector pET28a. The recombinant smASL protein was expressed in a soluble form in Escherichia coli strain BL21 (DE3) and purified to homogeneity by two-step column chromatography. Crystals suitable for X-ray analysis were obtained and X-ray diffraction data were collected to a resolution of 2.5 A. The crystals belonged to space group R3, with unit-cell parameters a = b = 254.5, c = 78.3 A. PMID- 21636912 TI - Crystallization and preliminary X-ray crystallographic analysis of dihydrouridine synthase from Thermus thermophilus and its complex with tRNA. AB - Dihydrouridine synthase (Dus) is responsible for catalyzing dihydrouridine formation in RNA by the reduction of uridine. To elucidate its RNA-recognition mechanism, Dus from Thermus thermophilus (TthDus) and its complex with tRNA were crystallized. Diffraction data sets were collected from crystals of native and selenomethionine-substituted TthDus to resolutions of 1.70 and 2.30 A, respectively. These crystals belonged to space group P1. Preliminary X-ray crystallographic analysis showed that two molecules of TthDus were contained in an asymmetric unit. In addition, diffraction data were collected to 3.51 A resolution from a crystal of selenomethionine-substituted TthDus in complex with tRNA, which belonged to space group P4(1)2(1)2. Preliminary structural analysis showed that the asymmetric unit contained two TthDus-tRNA complexes. PMID- 21636913 TI - Cloning, expression, purification, crystallization and X-ray crystallographic analysis of glucuronic acid dehydrogenase from Chromohalobacter salexigens. AB - Glucuronic acid dehydrogenase (GluUADH), the product of the Csal-2474 gene from the halophilic bacterium Chromohalobacter salexigens DSM 3043, is an enzyme with potential use in the conversion of glucuronic acid in seaweed biomass to fuels and chemicals. GluUADH is an enzyme that catalyzes the oxidation of glucuronic acid (GluUA) and galacturonic acid (GalUA) and has a preference for NAD(+) rather than NADP(+) as a cofactor. Recombinant GluUADH was crystallized in the presence of 0.2 M calcium acetate, 0.1 M Tris-HCl pH 7.0 and 20% PEG 3000 at 295 K. X-ray diffraction data were collected to a maximum resolution of 2.1 A. The GluUADH crystal belonged to space group P6(3), with unit-cell parameters a = b = 122.58, c = 150.49 A, gamma = 120 degrees . With one molecule per asymmetric unit, the crystal volume per unit protein weight (V(M)) is 2.78 A(3) Da(-1). The structure was solved by the single anomalous dispersion method and structure refinement is in progress. PMID- 21636914 TI - Cloning, expression, purification, crystallization and preliminary X-ray crystallographic study of GK0767, the copper-containing nitrite reductase from Geobacillus kaustophilus. AB - The soluble region (residues 32-354) of GK0767, a copper-containing nitrite reductase from the thermophilic Gram-positive bacterium Geobacillus kaustophilus HTA426, has been cloned and overexpressed in Escherichia coli. The purified recombinant protein was crystallized using the hanging-drop vapour-diffusion method. X-ray diffraction data were collected and processed to a maximum resolution of 1.3 A. The crystals belonged to space group R3, with unit-cell parameters a = b = 115.1, c = 87.5 A. Preliminary studies and molecular replacement calculations reveal the presence of one subunit of the homotrimeric structure in the asymmetric unit; this corresponds to a V(M) value of 3.14 A(3) Da(-1). PMID- 21636915 TI - Crystallization and preliminary X-ray crystallographic analysis of the human kindlin-2 PH domain. AB - Kindlins contribute to the correct assembly of integrin-containing focal adhesion sites through their direct interaction with the cytoplasmic tail of beta integrin. The FERM domain of kindlins has a unique subdomain organization: the F2 subdomain harbours a centrally located pleckstrin homology (PH) domain that is thought to be involved in the membrane targeting of kindlins. FERM domains are found in a number of cytoskeletal proteins that mediate the interaction between integrins and cytosolic proteins. In the present study, the PH domain of human kindlin-2 was subcloned, solubly expressed in Escherichia coli and crystallized using the hanging-drop vapour-diffusion method. A diffraction data set was collected at 2.8 A resolution using synchrotron radiation on BL-4A at the Pohang Accelerator Laboratory (Pohang, Republic of Korea). PMID- 21636916 TI - Crystallization and preliminary X-ray studies of a galactose-specific lectin from the seeds of Spatholobus parviflorus. AB - A galactose-specific seed lectin was purified from the legume Spatholobus parviflorus and crystallized using the hanging-drop vapour-diffusion technique. The crystals belonged to space group P1, with unit-cell parameters a = 60.998, b = 60.792, c = 78.179 A, alpha = 101.32, beta = 91.38, gamma = 104.32 degrees . X ray diffraction data were collected under cryoconditions (100 K) to a resolution of 2.04 A using a MAR image-plate detector system mounted on a rotating-anode X ray (Cu Kalpha) generator. Molecular replacement using legume-lectin coordinates as a search model gave a tetrameric structure. PMID- 21636917 TI - Crystallization and preliminary X-ray diffraction analysis of kanamycin-binding beta-lactamase in complex with its ligand. AB - TEM-1 beta-lactamase is a highly efficient enzyme that is involved in bacterial resistance against beta-lactam antibiotics such as penicillin. It is also a robust scaffold protein which can be engineered by molecular-evolution techniques to bind a variety of targets. One such beta-lactamase variant (BlaKr) has been constructed to bind kanamycin (kan) and other aminoglycoside antibiotics, which are neither substrates nor ligands of native beta-lactamases. In addition to recognizing kan, BlaKr activity is up-regulated by its binding via an activation mechanism which is not yet understood at the molecular level. In order to fill this gap, determination of the structure of the BlaKr-kan complex was embarked upon. A crystallization condition for BlaKr-kan was identified using high throughput screening, and crystal growth was further optimized using streak seeding and hanging-drop methods. The crystals belonged to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 47.01, b = 72.33, c = 74.62 A, and diffracted to 1.67 A resolution using synchrotron radiation. The X-ray structure of BlaKr with its ligand kanamycin should provide the molecular-level details necessary for understanding the activation mechanism of the engineered enzyme. PMID- 21636918 TI - Crystallization and preliminary crystallographic studies of Helicobacter pylori arginase. AB - Helicobacter pylori arginase is an important factor in evasion of the host's immune system and contributes to persistent infection by this bacterium. It is unique in many aspects compared with other arginases: for example, it has optimal activity with Co(2+) as a cofactor rather than Mn(2+) and has strongest activity at acidic pH instead of alkaline pH. In this study, H. pylori arginase was purified and crystallized in complex with Mn(2+) and a diffraction data set was collected to 2.2 A resolution. The crystals belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 94.69, b = 102.24, c = 148.61 A. PMID- 21636919 TI - Crystallization and preliminary X-ray characterization of the glpX-encoded class II fructose-1,6-bisphosphatase from Mycobacterium tuberculosis. AB - Fructose-1,6-bisphosphatase (FBPase; EC 3.1.3.11), which is a key enzyme in gluconeogenesis, catalyzes the hydrolysis of fructose 1,6-bisphosphate to form fructose 6-phosphate and orthophosphate. The present investigation reports the crystallization and preliminary crystallographic studies of the glpX-encoded class II FBPase from Mycobacterium tuberculosis H37Rv. The recombinant protein, which was cloned using an Escherichia coli expression system, was purified and crystallized using the hanging-drop vapor-diffusion method. The crystals diffracted to a resolution of 2.7 A and belonged to the hexagonal space group P6(1)22, with unit-cell parameters a = b = 131.3, c = 143.2 A. The structure has been solved by molecular replacement and is currently undergoing refinement. PMID- 21636920 TI - Purification, crystallization and X-ray characterization of a Kunitz-type trypsin inhibitor protein from the seeds of chickpea (Cicer arietinum). AB - A Kunitz-type trypsin inhibitor protein (CPTI) purified from chickpea seeds was estimated to have a molecular mass of 18 kDa on SDS-PAGE. The IC(50) value of CPTI was determined to be 2.5 ug against trypsin. The inhibitory activity of CPTI is 114 TIU (trypsin inhibitory units) per milligram of protein, which is high compared with those of other known Kunitz-type trypsin inhibitors from legumes. CPTI crystallized in three different orthorhombic crystal forms: P2(1)2(1)2 form A, P2(1)2(1)2 form B and P2(1)2(1)2(1). The crystals of P2(1)2(1)2 form A, with unit-cell parameters a = 37.2, b = 41.2, c = 104.6 A, diffracted to 2.0 A resolution at the home source and to 1.4 A on beamline BM14 at the ESRF. Data were also collected from crystals grown in the presence of iodine. The Matthews coefficient for these crystals was calculated to be 2.37 A(3) Da(-1), corresponding to a solvent content of 42%. The other two crystal forms (P2(1)2(1)2 form B and P2(1)2(1)2(1)) diffracted comparatively poorly. PMID- 21636921 TI - Crystallization and preliminary structural analysis of the Listeria monocytogenes Ca(2+)-ATPase LMCA1. AB - Ca(2+)-ATPases are ATP-driven membrane pumps that are responsible for the transport of Ca(2+) ions across the membrane. The Listeria monocytogenes Ca(2+) ATPase LMCA1 has been crystallized in the Ca(2+)-free state stabilized by AlF(4)( ), representing an occluded E2-P(i)-like state. The crystals belonged to space group P2(1)2(1)2 and a complete data set extending to 4.3 A resolution was collected. A molecular-replacement solution was obtained, revealing type I packing of the molecules in the crystal. Unbiased electron-density features were observed for AlF(4)(-) and for shifts of the helices, which were indicative of a reliable structure determination. PMID- 21636922 TI - Expression, crystallization and preliminary X-ray analysis of a ferric binding protein from Thermus thermophilus HB8. AB - A ferric binding protein from Thermus thermophilus HB8 (TtFbpA) was expressed, purified and crystallized using the hanging-drop vapour-diffusion method. Four different crystal forms were obtained and characterized by X-ray diffraction. Two crystal forms with TtFbpA in the apo state belonged to the orthorhombic space group P2(1)2(1)2(1) (unit-cell parameters a = 42.1, b = 139.3, c = 326.5 A and a = 42.1, b = 139.3, c = 218.9 A). The third form with TtFbpA also in the apo state belonged to the monoclinic space group P2(1) (unit-cell parameters a = 66.5, b = 61.7, c = 73.9 A, beta = 111.7 degrees ). The fourth form, with TtFbpA in the iron-bound holo state as confirmed by an atomic absorption spectrophotometry assay, belonged to the trigonal space group P3(1)21 or P3(2)21 (unit-cell parameters a = 63.6, b = 63.6, c = 266.7 A, alpha = beta = 90.0, gamma = 120.0 degrees ). PMID- 21636923 TI - Crystallization and preliminary crystallographic studies of UbiG, an O methyltransferase from Escherichia coli. AB - UbiG, an O-methyltransferase from the ubiquinone-biosynthesis pathway in Escherichia coli, catalyzes two O-methyl transfer steps. The primary structures of the O-methyltransferase enzyme family used in ubiquinone synthesis are conserved in both prokaryotes and eukaryotes, but their tertiary structures and catalytic mechanisms are not yet known. Here, UbiG with an N-terminal hexahistidine tag was expressed and crystallized. Crystals grown by the hanging drop vapour-diffusion method diffracted to 2.00 A resolution and belonged to space group C121, with unit-cell parameters a = 119.8, b = 58.6, c = 40.2 A, beta = 105.3 degrees . Both Matthews coefficient analysis and the self-rotation function suggested the presence of one molecule per asymmetric unit in the crystal, with a solvent content of 50.52% (V(M) = 2.48 A(3) Da(-1)). PMID- 21636924 TI - Crystallization and preliminary X-ray crystallographic analysis of eIF5BDeltaN and the eIF5BDeltaN-eIF1ADeltaN complex. AB - The binding between two universally conserved translation initiation factors, eIF5B and eIF1A, is important in the initiation step of eukaryotic protein synthesis on the ribosome. Through this interaction, eIF1A assists in recruiting eIF5B to the initiating 40S subunit; eIF5B then encourages the joining of the 60S subunit to form an initiating 80S ribosome. Here, the expression, purification, crystallization and preliminary X-ray analyses of eIF5BDeltaN and the eIF5BDeltaN eIF1ADeltaN complex from Saccharomyces cerevisiae are reported. The crystal of eIF5BDeltaN diffracted to 2.45 A resolution and belonged to space group P4(1)2(1)2, with unit-cell parameters a = b = 130.0, c = 71.7 A. The asymmetric unit was estimated to contain one molecule. The initial phase was obtained by Se SAD. The crystal of the eIF5BDeltaN-eIF1ADeltaN complex diffracted to 3.3 A resolution and belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 101.9, b = 120.9, c = 132.8 A. The asymmetric unit was estimated to contain two complex molecules. PMID- 21636925 TI - Pediatric cardiac program in India: changing perspectives. PMID- 21636926 TI - Cardiac tamponade and clinical acumen. PMID- 21636927 TI - Validation of a decision-making strategy for systolic anterior motion following mitral valve repair. AB - Low cardiac output syndrome and hypotension are dreadful consequences of systolic anterior motion (SAM) after a mitral valve (MV) repair. The management of SAM in the operating room remains controversial. We validate a recently suggested two step management method and classification of this complication. This was a teaching hospital-based observational study. We validated a novel two-step conservative management method, consisting in intravascular volume expansion and discontinuation of inotropic drugs (step 1), and increasing the afterload by ascending aorta manual compression while administering esmolol e.v. (step 2). We also validate a novel classification of SAM: easy-to-revert (responding to step 1), difficult-to-revert (responding to step 2), or persistent. Fifty patients had an easy-to-revert while 26 had a difficult-to-revert SAM; 4 patients had a persistent condition (promptly diagnosed through our decisional algorithm) and underwent an immediate second pump run to repeat the mitral repair surgery. We confirmed that SAM after a repair of a degenerative MV is common and validated a simple two-step conservative management method that allows to clearly identify those few patients who require immediate surgical revision. PMID- 21636928 TI - Real-time three-dimensional echocardiographic assessment of mitral valve: Is it really superior to 2D transesophageal echocardiography? AB - Aim of our study was to investigate the feasibility of use and possible additional value of real-time 3D transesophageal echocardiography (RT-3D-TEE) compared to conventional 2D-TEE in patients undergoing elective mitral valve repair. After ethical committee approval, patients were included in this prospective study. After induction of anesthesia, a comprehensive 2D-TEE examination was performed, followed with RT-3D-TEE. The intraoperative surgical finding was used as the gold standard for segmental analysis. Only such segments which were surgically corrected either by resection or insertion of artificial chords were judged pathologic. A total of 50 patients were included in this study; usable data were available from 42 of these patients . Based on the Carpentier classification, the pathology found was type I in 2 (5%) patients, type II in 39 (93%) patients and type IIIb in 1 (2%) patient. We found that 3D imaging of complex mitral disease involving multiple segments, when compared to 2D-TEE did not show any statistically significant difference.RT-3D-TEE did not show any major advantage when compared to conventional 2D-TEE for assessing mitral valve pathology, although further study in a larger population is required to establish the validity of this study. PMID- 21636929 TI - Anesthetic management of patients undergoing extra-anatomic renal bypass surgery for renovascular hypertension. AB - Renal artery disease is the most common cause for surgically curable form of hypertension. In a small subset of patients with severe aortic disease where the aorta is not suitable for endovascular technique and to provide an arterial inflow, an extra-anatomic renal bypass surgery (EARBS) is an option. Anesthetic management of such procedures has not been described so far in the literature. We retrospectively analyzed the anesthetic techniques used in all patients who underwent EARBS between February 1998 and June 2008 at this institute. We also further analyzed data concerning blood pressure (BP) control and renal function response following surgery as outcome variable measures. A total of 11 patients underwent EARBS during this period. Five received oral clonidine with premedication. During laryngoscopy, esmolol was used in 4 patients, while lignocaine was used in remaining 7 patients. Of 11 patients, 7 showed significant hemodynamic response to laryngoscopy and intubation; among these, one had oral clonidine with premedicant, and 6 received lignocaine just before laryngoscopy. Intravenous vasodilators were used to maintain target BP within 20% of baseline during perioperative period. All patients received renal protective measures. During follow-up, 10% were considered cured, 70% had improved BP response, while 20% failed to show improvement in BP response. Renal functions improved in 54.5%, remain unchanged in 36.5%, and worsened in 9% of patients. Use of clonidine during premedication and esmolol before laryngoscopy were beneficial in attenuating hemodynamic response to laryngoscopy, while use of vasodilators to maintain target BP within 20% of baseline, and routine use of renal protective measures appear to be promising in patients undergoing EARBS. PMID- 21636930 TI - Comparison of transthoracic electrical bioimpedance cardiac output measurement with thermodilution method in post coronary artery bypass graft patients. AB - Transthoracic electrical bioimpedance (TEB) has been proposed as a non-invasive, continuous, and cost-effective method of cardiac output (CO) measurement. In this prospective, non-randomized, clinical study, we measured CO with NICOMON (Larsen and Toubro Ltd., Mysore, India) and compared it with thermodilution (TD) method in patients after off-pump coronary artery bypass (OPCAB) graft surgery. We also evaluated the effect of ventilation (mechanical and spontaneous) on the measurement of CO by the two methods. Forty-six post-OPCAB patients were studied at five predefined time points during controlled ventilation and at five time points when breathing spontaneously. A total of 230 data pairs of CO were obtained. During controlled ventilation, TD CO values ranged from 2.29 to 6.74 L/min (mean 4.45 +/- 0.85 L/min), while TEB CO values ranged from 1.70 to 6.90 L/min (mean 4.43 +/- 0.94 L/min). The average correlation (r) was 0.548 (P = 0.0002), accompanied by a bias of 0.015 L/min and precision of 0.859 L/min. In spontaneously breathing patients, TD CO values ranged from 2.66 to 6.92 L/min (mean 4.66 +/- 0.76 L/min), while TEB CO values ranged from 3.08 to 6.90 L/min (mean 4.72 +/- 0.82 L/min). Their average correlation was relatively poor (r = 0.469, P= 0.002), accompanied by a bias of -0.059 L/min and precision of 0.818 L/min. The overall percent errors between TD CO and TEB CO were 19.3% (during controlled ventilation) and 17.4% (during spontaneous breathing), respectively. To conclude, a fair correlation was found between TD CO and TEB CO measurements among post-OPCAB patients during controlled ventilation. However, the correlation was weak in spontaneously breathing patients. PMID- 21636931 TI - Pericardial tamponade after left posterolateral thoracotomy for left upper lobectomy for pulmonary aspergilloma. AB - Pericardial tamponade limits diastolic filling of the heart; therefore, a high venous pressure is required to fill the ventricle. In presence of cardiac tamponade, therapeutic agents and manoeuvres that results in venodilation or vasodilation can severely compromise diastolic filling of the heart and might result in rapid cardiac decompensation. Equalization of central venous pressure and pulmonary artery diastolic pressure or equalization of pressures in all four chambers during diastole confirms cardiac tamponade. Transthoracic echocardiography can detect the site of tamponade and assist in pericardiocentesis. We describe acute pericardial tamponade in a young man who underwent left posterolateral thoracotomy for left upper lobectomy. Intraoperatively, mobilization of the left upper lobe was frequently associated with hypotension. Postoperatively, the patient suffered two more episodes of hypotension. The episodes of hypotension were attributed to surgical manipulation and epidural blockade. Hemodynamics normalized after discontinuing epidural infusion, volume resuscitation and lobectomy. On third postoperative day, the patient developed cardiovascular collapse; arterial blood pressure and central venous pressure were 70/50 and 12 mmHg. Investigations showed haziness of left lung, and severe respiratory acidosis. On opening of the left thoracotomy wound, pericardial tamponade was diagnosed. A pericardial window was created and tamponade was released with that the hemodynamics normalized. Episodes of unexplained hypotension after left upper lobectomy suggest a cardiac etiology and acute pericardial tamponade is a possibility which should be released immediately otherwise it can result in fatal outcome. PMID- 21636932 TI - Anesthetic management for combined mitral valve replacement and aortic valve repair in a patient with osteogenesis imperfecta. AB - Osteogenesis imperfecta is a rare disorder of connective tissues and presents multiple challenges, including difficult airway, hyperthermia, coagulopathy and respiratory dysfunction, for anesthesiologists, especially during cardiac surgery. We present anesthetic management of a patient with osteogenesis impertecta during double valve surgery. Dexmedetomidine infusion minimized the risks of malignant hyperthermia. Glidescope and in-line stabilization facilitated endotracheal intubation and protected his oral structures and cervical spine. Transesophageal echocardiography (TEE) diagnosed a flail A3 segment and redundant left coronary cusp causing mitral and aortic regurgitation. The mitral valve was replaced and the aortic valve repaired. Coagulopathy was corrected according to comprehensive coagulation analysis. Glidescope, dexmedetomidine, coagulation analysis and TEE could facilitate anesthetic management in these patients. PMID- 21636933 TI - An unknown complication of peripherally inserted central venous catheter in a patient with ventricular assist device. AB - We report an unknown complication of peripherally inserted central venous catheter in a patient with Ventricular Assist Device. This rare complication led to the failure of the right ventricular assist device, which could be detrimental in patients with dilated cardiomyopathy. PMID- 21636934 TI - Placement of an implantable cardioverter-defibrillator in an infant with congenital long QT syndrome: anesthetic considerations. AB - Sudden cardiac arrest (SCA) in children is a rare, but catastrophic event. Children with cardiac pathology at particular risk include those with congenital long QT syndrome (CLQTS) and hypertrophic cardiomyopathy. CLQTS is a genetic disorder of the cardiac ion channels and is associated with significant risk of malignant ventricular arrhythmias and SCA. For symptomatic, untreated patients, the mortality rate is approximately 20% for the first year and 50% at ten years. Use of an implantable cardioverter-defibrillator (ICD) is recommended for the prevention of SCA in this patient population. We report a case of CLQTS, who after successful resuscitation from SCA, underwent ICD placement at our center. PMID- 21636935 TI - Perioperative hypotension and myocardial ischemia: diagnostic and therapeutic approaches. AB - Although perioperative hypotension is a common problem, its true incidence is largely unknown. There is evidence that postoperative outcome, including the incidence of myocardial adverse events, may be linked to the prolonged episodes of perioperative hypotension. Despite this, there are very few comprehensive resources available in the literature regarding diagnosis and management of these not so uncommon clinical occurrences, especially during non-cardiac surgery. Most anesthesia providers consider intraoperative hypotension to be caused by systemic vasodilatation and relative hypovolemia and so treat it empirically. The introduction of new monitoring devices including transesophageal echocardiography and arterial pressure waveform based stroke volume measurement have provided additional tools to narrow the differential diagnoses and initiate optimal treatment measures. Understanding the basic pathophysiology of hypotension and myocardial ischemia can further assist in providing goal directed management. This article serves as a comprehensive guide for anesthesiologists to diagnose and treat hypotension and myocardial ischemia. A summary of available techniques to monitor perioperative myocardial ischemia and their limitations are also discussed. PMID- 21636936 TI - Role of multimodality cardiac imaging in preoperative cardiovascular evaluation before noncardiac surgery. AB - The preoperative cardiac assessment of patients undergoing noncardiac surgery is common in the daily practice of medical consultants, anesthesiologists, and surgeons. The number of patients undergoing noncardiac surgery worldwide is increasing. Currently, there are several noninvasive diagnostic tests available for preoperative evaluation. Both nuclear cardiology with myocardial perfusion single photon emission computed tomography (SPECT) and stress echocardiography are well-established techniques for preoperative cardiac evaluation. Recently, some studies demonstrated that both coronary angiography by gated multidetector computed tomography and stress cardiac magnetic resonance might potentially play a role in preoperative evaluation as well, but more studies are needed to assess the role of these new modalities in preoperative risk stratification. A common question that arises in preoperative evaluation is if further preoperative testing is needed, which preoperative test should be used. The preferred stress test is the exercise electrocardiogram (ECG). Stress imaging with exercise or pharmacologic stress agents is to be considered in patients with abnormal rest ECG or patients who are unable to exercise. After reviewing this article, the reader should develop an understanding of the following: (1) the magnitude of the cardiac preoperative morbidity and mortality, (2) how to select a patient for further preoperative testing, (3) currently available noninvasive cardiac testing for the detection of coronary artery disease and assessment of left ventricular function, and (4) an approach to select the most appropriate noninvasive cardiac test, if needed. PMID- 21636937 TI - Antiphospholipid syndrome, cardiac surgery and cardiopulmonary bypass. PMID- 21636938 TI - Left juxtaposed atrial appendages in a patient with dextrocardia and tricuspid atresia: TEE images. PMID- 21636939 TI - Unmasking of patent ductus arteriosus on cardiopulmonary bypass: role of intraoperative trans-esophageal echocardiography in a patient with severe pulmonary hypertension due to pulmonary vein stenosis and cor triatriatum. PMID- 21636940 TI - A simple method of electrocardiogram: controlled central venous catheterization. PMID- 21636941 TI - Dengue fever in a patient recovering from coronary artery bypass grafting. PMID- 21636942 TI - Post-extubation pulmonary edema after open cholecystectomy: significance of diastolic cardiac dysfunction. PMID- 21636943 TI - Bipolar hip arthroplasty in an adult patient with uncorrected tetralogy of fallot: anesthetic management. PMID- 21636944 TI - Heart and ECMO: are we ready? PMID- 21636945 TI - Comparison of cardiac output estimation by FloTrac/Vigileo TM and intermittent pulmonary artery thermodilution in patient with Takayasu arteritis. PMID- 21636946 TI - Artifactual ST segment depression induced by electrocautery. PMID- 21636947 TI - Pulsus alternans after aortic valve replacement: a preventable yet a possible risk of cardiac manipulation - fact or fiction? PMID- 21636948 TI - To evaluate the heart or not in emergency neurosurgical head-injured patients with ST elevation: authors' reply. PMID- 21636949 TI - C-reactive protein (CRP) is not a mere marker, but an active pathogenic substance. PMID- 21636951 TI - Effect of linear polarized near-infrared light irradiation and light exercise on muscle performance. AB - This study aimed to determine the effect of active warm-up by local muscle light exercise and passive warm-up by polarized light irradiation on skin and muscle temperatures and forearm muscle performance (muscle strength, power, endurance, and controlled force-exertion). Ten healthy males performed various grip tests before and after active (local muscle light exercise) and passive (linear polarized near-infrared light irradiation) warm-ups. An active warm-up involved intermittent gripping exercise (contraction: 1 second and relaxation: 1 second) for 10 minutes using a sponge. A passive warm-up consisted of polarized light irradiation to the forearm (superficial digital flexor) for 10 minutes (irradiation: 5 seconds and rest: 1 second). Skin and muscle temperatures were measured during both warm-ups. Skin and muscle temperatures increased significantly after 5 minutes of local muscle light exercise and after 10 minutes of polarized light irradiation. Temperatures were significantly higher after 6 minutes of local muscle light exercise than after 6 minutes of polarized light irradiation. There were no significant differences of muscle strength, power, and controlled force-exertion before and after either warm-up. Average force outputs in all conditions significantly decreased with exertion time, and at 30, 60, 90, and 120 seconds they were higher in both warm-up conditions than in the non-warm up condition. In conclusion, both warm-ups may contribute to improve muscle endurance performance in the decreasing force phase. PMID- 21636950 TI - Effect of sulfonylurea agents on reverse cholesterol transport in vitro and vivo. AB - AIM: Reverse cholesterol transport (RCT) is a critical mechanism for the anti atherogenic property of HDL. The inhibitory effect of the sulfonylurea agent (SUA) glibenclamide on ATP binding-cassette transporter (ABC) A1 may decrease HDL function but it remains unclear whether it attenuates RCT in vivo. We therefore investigated how the SUAs glibenclamide and glimepiride affected the functionality of ABCA1/ABCG1 and scavenger receptor class B type I (SR-BI) expression in macrophages in vitro and overall RCT in vivo. METHODS: RAW264.7, HEK293 and BHK-21 cells were used for in vitro studies. To investigate RCT in vivo, 3H-cholesterol-labeled and acetyl LDL-loaded RAW264.7 cells were injected into mice. RESULTS: High dose (500uM) of glibenclamide inhibited ABCA1 function and apolipoprotein A-I (apoA-I)-mediated cholesterol efflux, and attenuated ABCA1 expression. Although glimepiride maintained apoA-I-mediated cholesterol efflux from RAW264.7 cells, like glibenclamide, it inhibited ABCA1-mediated cholesterol efflux from transfected HEK293 cells. Similarly, the SUAs inhibited SR-BI mediated cholesterol efflux from transfected BHK-21 cells. High doses of SUAs increased ABCG1 expression in RAW264.7 cells, promoting HDL-mediated cholesterol efflux in an ABCG1-independent manner. Low doses (0.1-100 uM) of SUAs did not affect cholesterol efflux from macrophages despite dose-dependent increases in ABCA1/G1 expression. Furthermore, they did not change RCT or plasma lipid levels in mice. CONCLUSION: High doses of SUAs inhibited the functionality of ABCA1/SR BI, but not ABCG1. At lower doses, they had no unfavorable effects on cholesterol efflux or overall RCT in vivo. These results indicate that SUAs do not have adverse effects on atherosclerosis contrary to previous findings for glibenclamide. PMID- 21636952 TI - Effects of different light intensities in the morning on dim light melatonin onset. AB - The present study evaluated the effects of exposure to light intensity in the morning on dim light melatonin onset (DLMO). The tested light intensities were 750 lux, 150 lux, 3000 lux, 6000 lux and 12,000 lux (horizontal illuminance at cornea), using commercial 5000 K fluorescent lamps. Eleven healthy males aged 21 31 participated in 2-day experiments for each light condition. On the first experimental day (day 1), subjects were exposed to dim light (<30 lux) for 3 h in the morning (09:00-12:00). On the same day, saliva samples were taken in dim light (<30 lux) every 30 min from 21:00 to 01:00 to determine the DLMO phase. The subjects were allowed to sleep from 01:00 to 08:00. On the second experimental day (day 2), the subjects were exposed to experimental light conditions for 3 h in the morning. The experimental schedule after light exposure was the same as on day 1. On comparing day 2 with day 1, significant phase advances of DLMO were obtained at 3000 lux, 6000 lux and 12,000 lux. These findings indicate that exposure to a necessary intensity from an ordinary light source, such as a fluorescent lamp, in the morning within one day affects melatonin secretion. PMID- 21636953 TI - Automatic sleep/wake scoring from body motion in bed: validation of a newly developed sensor placed under a mattress. AB - The purpose of this study was to formulate a "sleep/wake" scoring algorithm for processing activity measurements obtained using a newly developed nonwear actigraphy (NWA) device, and to test its validity. The NWA device has a highly sensitive pressure sensor and is placed under a mattress. It can continuously record the activity of a person lying on the mattress and identify an "in-bed/out of-bed" state from the vibrations of the mattress. We formulated the sleep/wake scoring algorithm by using data obtained simultaneously by wrist actigraphy (Act) and the NWA device in 33 healthy participants. Agreement rate, sensitivity, and specificity with Act were 95.7%, 97.6%, and 75.8% (33 healthy people); the corresponding values were 85.9%, 89.1%, and 79.8% for 12 nursing home residents and 93.7%, 97.2%, and 60.8% for 60 nights for 6 healthy persons who slept 10 nights on their futons. Agreement rate, sensitivity, and specificity with polysomnography were in almost perfect agreement with Act (12 nights; 6 healthy persons who slept 2 nights). All our validation results indicate that the NWA device, placed under a mattress or a futon, can produce almost identical sleep/wake scores to Act. It is expected that the NWA device, a nonwear device for scoring sleep/wake and in-bed/out-of-bed, enables convenient long-term sleep related evaluation in various fields, including hospital settings, home-care settings, and care facility settings such as nursing homes. PMID- 21636954 TI - Serum leptin concentration and its effect on puberty in Naqu Tibetan adolescents. AB - This study aimed to clarify the regularity of leptin in Naqu Tibetan adolescents. This study investigated the concentration of fasting serum leptin and clarified its relationship between BMI and other indices. Healthy Naqu Tibetan adolescents aged 12-18 were investigated randomly in the study. They were divided into seven groups (each year as one group, 12 boys and 12 girls in each group); serum concentrations of leptin, estradiol, testosterone (T), follicle stimulating hormone (FSH) and luteinizing hormone (LH) were analyzed. The height and body weight of the 168 healthy Naqu Tibetan adolescents were also assessed. The leptin level in boys decreased with age but increased in girls; in boys and girls they both differed between groups (p<0.05). In boys, the leptin level was inversely correlated with body mass index (BMI), FSH, and T (p<0.05), while in girls, it was positively related to BMI, FSH, LH, and E2 (p<0.01).These findings suggested that during puberty the serum leptin concentration increased with age in girls while it decreased in boys; in the same age group, the leptin level in girls was significantly higher than in boys. Leptin may have some relationship with puberty in Tibetan adolescents. PMID- 21636955 TI - Effects of walking speed and step frequency on estimation of physical activity using accelerometers. AB - This study evaluated the accuracy of assessing step counts and energy costs under walking conditions altered by step frequency changes at given speeds using uni- (LC) and tri-axial accelerometers (AM, ASP). Healthy young men and women (n=18) volunteered as subjects. Nine tests were designed to manipulate three step frequencies, low (-15% of normal), normal, and high (+15%), at each walking speed (55, 75, and 95 m/min). A facemask connected to a Douglas bag was attached to subjects, who wore accelerometers around their waist. LC underestimated the step counts at normal or high step frequency at 55 m/min and AM also at all step frequencies at 55 m/min, whereas ASP did not in all trials. LC underestimated metabolic equivalents (METs) at low or normal step frequency at all walking speeds. AM underestimated METs at low step frequency at all walking speeds and at high step frequency of 95 m/min. ASP gave underestimates only at low step frequency of 95 m/min. The degree of the percentage error of METs for AM and ASP was affected by step frequency. Significant interaction between step frequency and speed was found that for LC. These results suggest that LC and AM can cause errors in step-count functions at a low walking speed. Furthermore, LC may show low accuracy of the METs measurement during walking altered according to step frequency and speed, whereas AM and ASP, which are tri-axial accelerometers, are more accurate but the degree of the percentage error is affected by step frequency. PMID- 21636956 TI - Non-CpG methylation occurs in the regulatory region of the Sry gene. AB - The Sry (sex determining region on Y chromosome) gene is a master gene for sex determination. We previously reported that the Sry gene has tissue-dependent and differentially methylated regions (T-DMRs) by analyzing the DNA methylation states at CpG sites in the promoter regions. In this study, we found unique non CpG methylation at the internal cytosine in the 5'-CCTGG-3' pentanucleotide sequence in the Sry T-DMR. This non-CpG methylation was detected in four mouse strains (ICR, BALB/c, DBA2 and C3H), but not in two strains (C57BL/6 and 129S1), suggesting that the CCTGG methylation is tentative and unstable. Interestingly, this CCTGG methylation was associated with demethylation of the CpG sites in the Sry T-DMR in the developmental process. A methylation-mediated promoter assay showed that the CCTGG methylation promotes gene expression. Our finding shows that non-CpG methylation has unique characteristic and is still conserved in mammals. PMID- 21636957 TI - Effect of fluorescent mercury light irradiation on in vitro and in vivo development of mouse oocytes after parthenogenetic activation or sperm microinjection. AB - The detection of specific cellular components using fluorescent agents such as green fluorescent protein (GFP), red fluorescent protein or Hoechst dyes provides a powerful tool for studying cell biology. However, specimens must be exposed to high-intensity light, which might cause cellular damage. Here, we exposed mouse metaphase stage (M) II oocytes to fluorescent mercury vapor light at three wavelengths (539 nm, 488 nm and 341 nm) to determine the maximum exposure time that would avoid damage. When oocytes were activated parthenogenetically after exposure to these wavelengths for more than 20 min, 5 min or 4 sec, respectively, the percentages of dead oocytes after activation increased, and none of the surviving embryos developed to blastocysts. However, embryos fertilized by intracytoplasmic sperm injection (ICSI) were more tolerant to light damage, even though the quality of blastocysts, judged by cell number and cell allocation to the inner cell mass and trophectoderm measured by immunostaining for Oct4 and Cdx2, was reduced as exposure times increased. Live, healthy offspring were obtained when these exposed embryos were transferred into recipient pseudopregnant females at the 2-cell stage. In addition, MII oocytes collected from GFP-expressing transgenic mice after 5 min of irradiation with 488-nm light were also able to develop to full term following ICSI. Thus, we determined the safe period of exposure to several wavelengths for oocyte manipulation or observation that would permit subsequent development. PMID- 21636960 TI - Insulin responses to administrations of amino acids and fatty acids in healthy cats. AB - In order to compare the stimulation ability of insulin secretion, we determined changes in plasma glucose and insulin concentrations after intravenous administration of various amino acids and essential fatty acids in clinically healthy adult cats. Plasma glucose concentrations were within the normal ranges after injection of amino acids and fatty acids. Plasma insulin concentrations increased rapidly 2 to 4 min after injection of arginine, then decreased to the basal levels at 20 min in all five cats. Insulin peak responses were significantly greater in arginine injections than in normal saline (P<0.01). Areas under the curve (AUC) of plasma insulin concentrations from 0 to 10 min after injection of arginine were significantly larger than after injection of normal saline (P<0.01) and glucose (P<0.05). Increases in AUC of plasma insulin concentration from 0 to 60 min were observed after injection of arginine, leucine, alanine, and fat emulsion. Arginine had a strong insulinotropic effect, and leucine, alanine, and fatty acids had weak ones. Besides, valine, methionine, taurine and glutamine had no stimulant activity of insulin. Given the risk of glucose toxication and required time for testing, the intravenous arginine tolerance test may be useful for estimation of insulin responses in cats. PMID- 21636961 TI - Molecular genetic and pathogenic characterization of psittacid herpesvirus type 1 isolated from a captive galah (Eolophus roseicapillus) in Japan. AB - Psittacid herpesvirus type 1 (PsHV-1) was isolated from a captive galah (Eolophus roseicapillus) in Japan that was suspected of having Pacheco's disease (PD), an acute fatal disease in psittacine birds. PsHV-1 has been classified into four genotypes based on the UL16 gene sequence. In the present study, we investigated the genetic and pathogenic characteristics of the isolated virus, FOY-1, compared with a reference strain, RSL-1. The FOY-1 strain was classified into PsHV-1 genotype 2. The FOY-1 strain was found to be less pathogenic to budgerigars than RSL-1, which was classified as genotype 4 in an in vivo study. This is the first report regarding the classification of originally isolated PsHV-1 in Japan and its characterization by animal infection experiment. PMID- 21636963 TI - Japanese guideline for adult asthma. AB - Adult bronchial asthma (hereinafter, asthma) is characterized by chronic airway inflammation, reversible airway narrowing, and airway hyperresponsiveness. Long standing asthma induces airway remodeling to cause an intractable asthma. The number of patients with asthma has increased, while the number of patients who die from asthma has decreased (1.7 per 100,000 patients in 2009). The aim of asthma treatment is to enable patients with asthma to lead a healthy life without any symptoms. A partnership between physicians and patients is indispensable for appropriate treatment. Long-term management with agents and elimination of causes and risk factors are fundamental to asthma treatment. Four steps in pharmacotherapy differentiate mild to intensive treatments; each step includes an appropriate daily dose of an inhaled corticosteroid (ICS), varying from low to high doses. Long-acting beta(2) agonists (LABA), leukotriene receptor antagonists, and theophylline sustained-release preparation are recommended as concomitant drugs, while anti-IgE antibody therapy is a new choice for the most severe and persistent asthma. Inhaled beta(2) agonists, aminophylline, corticosteroids, adrenaline, oxygen therapy, etc., are used as needed against acute exacerbations. Allergic rhinitis, chronic obstructive pulmonary disease (COPD), aspirin induced asthma, pregnancy, and cough variant asthma are also important factors that need to be considered. PMID- 21636964 TI - Japanese guideline for childhood asthma. AB - The Japanese Guideline for the Diagnosis and Treatment of Allergic Diseases 2010 (JAGL 2010) describes childhood asthma based on the Japanese Pediatric Guideline for the Treatment and Management of Asthma 2008 (JPGL 2008) published by the Japanese Society of Pediatric Allergy and Clinical Immunology. JAGL 2010 provides information on diagnosis by age groups from infancy to puberty, treatment for acute exacerbations, long-term management by medication, daily life guidance, and patient education to allow physicians, not specialized in childhood asthma, to refer to this guideline for routine medical treatment. JAGL differs from the Global Initiative for Asthma Guideline (GINA) in that the former emphasizes long term management of childhood asthma based on asthma severity and early diagnosis and intervention at <2 years and 2-5 years of age. However, a management method, including step-up or step-down of long-term management agents based on the status of asthma symptoms, is easy to understand and thus JAGL is suitable for routine medical treatment. JAGL also introduced treatment and management using a control test for children, recommending treatment and management aimed at complete control through avoiding exacerbation factors and appropriate use of antiinflammatory agents. PMID- 21636965 TI - Japanese guideline for allergic rhinitis. AB - Like asthma and atopic dermatitis, allergic rhinitis is an allergic disease, but of the three, it is the only type I allergic disease. Allergic rhinitis includes pollinosis, which is intractable and reduces quality of life (QOL) when it becomes severe. A guideline is needed to understand allergic rhinitis and to use this knowledge to develop a treatment plan. In Japan, the first guideline was prepared after a symposium held by the Japanese Society of Allergology in 1993. The current 6th edition was published in 2009, and is widely used today. To incorporate evidence based medicine (EBM) introduced from abroad, the most recent collection of evidence/literature was supplemented to the Practical Guideline for the Management of Allergic Rhinitis in Japan 2009. The revised guideline includes assessment of diagnosis/treatment and prescriptions for children and pregnant women, for broad clinical applications. An evidence-based step-by-step strategy for treatment is also described. In addition, the QOL concept and cost benefit analyses are also addressed. Along with Allergic Rhinitis and its Impact of Asthma (ARIA), this guideline is widely used for various clinical purposes, such as measures for patients with sinusitis, childhood allergic rhinitis, oral allergy syndrome, and anaphylaxis and for pregnant women. PMID- 21636966 TI - Japanese guideline for allergic conjunctival diseases. AB - The definition, classification, pathogenesis, test methods, clinical findings, criteria for diagnosis, and therapies of allergic conjunctival disease are summarized based on the Guidelines for Clinical Management of Allergic Conjunctival Disease (Second Edition) revised in 2010. Allergic conjunctival disease is defined as "a conjunctival inflammatory disease associated with a Type I allergy accompanied by some subjective or objective symptoms." Allergic conjunctival disease is classified into allergic conjunctivitis, atopic keratoconjunctivitis, vernal keratoconjunctivitis, and giant papillary conjunctivitis. Representative subjective symptoms include ocular itching, hyperemia, and lacrimation, whereas objective symptoms include conjunctival hyperemia, swelling, folliculosis, and papillae. Patients with vernal keratoconjunctivitis, which is characterized by conjunctival proliferative changes called giant papilla accompanied by varying extents of corneal lesion, such as corneal erosion and shield ulcer, complain of foreign body sensation, ocular pain, and photophobia. In the diagnosis of allergic conjunctival diseases, it is required that type I allergic diathesis is present, along with subjective and objective symptoms accompanying allergic inflammation. The diagnosis is ensured by proving a type I allergic reaction in the conjunctiva. Given that the first-line drug for the treatment of allergic conjunctival disease is an antiallergic eye drop, a steroid eye drop will be selected in accordance with the severity. In the treatment of vernal keratoconjunctivitis, an immunosuppressive eye drop will be concomitantly used with the above mentioned drugs. PMID- 21636967 TI - Japanese guideline for atopic dermatitis. AB - Given the importance of appropriate diagnosis and appropriate assessment of cutaneous symptoms in treatment of atopic dermatitis, the basics of treatment in this guideline are composed of (1) investigation and countermeasures of causes and exacerbating factors, (2) correction of skin dysfunctions (skin care), and (3) pharmacotherapy, as three mainstays. These are based on the disease concept that atopic dermatitis is a inflammatory cutaneous disease with eczema by atopic diathesis, multi-factorial in onset and aggravation, and accompanied by skin dysfunctions. These three points are equally important and should be appropriately combined in accordance with the symptoms of each patient. In treatment, it is important to transmit the etiological, pathological, physiological, or therapeutic information to the patient to build a favorable partnership with the patient or his/her family so that they may fully understand the treatment. This guideline discusses chiefly the basic therapy in relation to the treatment of this disease. The goal of treatment is to enable patients to lead an uninterrupted social life and to control their cutaneous symptoms so that their quality of life (QOL) may meet a satisfactory level. PMID- 21636968 TI - Japanese guideline for food allergy. AB - Food allergy is defined as "a phenomenon in which adverse reactions (symptoms in skin, mucosal, digestive, respiratory systems, and anaphylactic reactions) are caused in living body through immunological mechanisms after intake of causative food." Various symptoms of food allergy occur in many organs. Food allergy falls into four general clinical types; 1) neonatal and infantile gastrointestinal allergy, 2) infantile atopic dermatitis associated with food allergy, 3) immediate symptoms (urticaria, anaphylaxis, etc.), and 4) food-dependent exercise induced anaphylaxis and oral allergy syndrome (i.e., specific forms of immediate type food allergy). Therapy for food allergy includes treatments of and prophylactic measures against hypersensitivity like anaphylaxis. A fundamental prophylactic measure is the elimination diet. However, elimination diets should be conducted only if they are inevitable because they places a burden on patients. For this purpose, it is highly important that causative foods are accurately identified. Many means to determine the causative foods are available, including history taking, skin prick test, antigen specific IgE antibodies in blood, basophil histamine release test, elimination diet test, oral food challenge test, etc. Of these, the oral food challenge test is the most reliable. However, it should be conducted under the supervision of experienced physicians because it may cause adverse reactions such as anaphylaxis. PMID- 21636969 TI - The duality of Aspergillus terreus: Differential immune responses to distinct conidia. PMID- 21636970 TI - Gibberellins negatively regulate low temperature-induced anthocyanin accumulation in a HY5/HYH-dependent manner. AB - Low temperature could significantly induce anthocyanin accumulation in the presence of light. Recently, two bZIP transcription factors LONG HYPOCOTYL 5 (HY5) and HOMOLOG OF HY5 (HYH) were identified to play an important role in the process of low temperature-induced anthocyanin accumulation. However, the mechanism by which HY5/HYH regulates anthocyanin accumulation under low temperature still remains unclear. Here, we found that the gibberellins (GAs) could decrease but PAC (endogenous GAs biosynthesis inhibitor) increase the low temperature-induced anthocyanin accumulation, implying that GAs signaling may involve in this process. Furthermore, the transcript levels of GA2ox1, encoding a major member of bioactive GAs-deactivating enzymes, were significantly up regulated by low temperature in a HY5/HYH-dependent manner. Moreover, hy5hyh mutant was insensitive to PAC in enhancing anthocyanin accumulation under low temperature. From these data we propose that, together with HY5/HYH, GA signaling may play an important role during low temperature-induced anthocyanin accumulation. PMID- 21636971 TI - There is more to autophagy than induction: regulating the roller coaster. AB - Considerable attention has been paid to the topic of autophagy induction. In part, this is because of the potential for modulating this process for therapeutic purposes. Of course we know that induced autophagy can also be problematic--for example, when trying to eliminate an established tumor that might be relying on autophagy for its own cytoprotective uses. Accordingly, inhibitory mechanisms have been considered; however, the corresponding studies have tended to focus on the pathways that block autophagy under non-inducing conditions, such as when nutrients are available. In contrast, relatively little is known about the mechanisms for inhibiting autophagy under inducing conditions. Yet, this type of regulation must be occurring on a routine basis. We know that dysregulation of autophagy, e.g., due to improper activation of Beclin 1 leading to excessive autophagy activity, can cause cell death. Accordingly, we assume that during starvation or other inducing conditions there must be a mechanism to modulate autophagy. That is, once you turn it on, you do not want to let it continue unchecked. But how is autophagy downregulated when the inducing conditions still exist? PMID- 21636972 TI - TRPM2 modulates insulin secretion in pancreatic beta-cells. AB - Insulin secretion from pancreatic beta-cells is the primary mechanism by which the body lowers blood glucose concentrations. Glucose is the principal stimulator of insulin secretion, and the primary pathway involved in glucose-stimulated insulin secretion is the ATP-sensitive K+ channel voltage-gated Ca2+ channel mediated pathway. Several TRP channels expressed in pancreatic beta-cells have been reported to be involved in insulin secretion. One recent report found that TRPM2 is expressed in pancreatic beta-cells and modulates insulin secretion stimulated by glucose and further potentiated by incretin hormones. TRPM2 is a Ca2+-permeable non-selective cation channel activated by adenosine dinucleotides, hydrogen peroxide, and intracellular Ca2+. Glucose tolerance was impaired and insulin secretion was decreased in TRPM2 knockout mice. Insulin secretion via TRPM2 occurs not only through control of intracellular Ca2+ concentrations but also through Ca2+ influx-independent mechanisms. Although further examination is needed to clarify the mechanism of TRPM2-mediated insulin secretion, TRPM2 may be a key player in regulation of insulin secretion and could represent a new target for diabetes therapy. PMID- 21636973 TI - DNA methylation in white blood cells: association with risk factors in epidemiologic studies. AB - Alterations in DNA methylation patterns, both at specific loci and overall in the genome, have been associated with many different health outcomes. In cancer and other diseases, most of these changes have been observed at the tissue level. Data on whether DNA methylation changes in white blood cells (WBC) can serve as a useful biomarker for different health outcomes are much more limited, but rapidly emerging. Epidemiologic studies have reported associations between global WBC methylation and several different cancers including cancers of the colon, bladder, stomach, breast and head and neck, as well as schizophrenia and myelodysplastic syndrome. Evidence for WBC methylation at specific loci and disease risk is more limited, but increasing. Differences in WBC DNA methylation by selected risk factors including demographic (age, gender, race), environmental exposures (benzene, persistent organic pollutants, lead, arsenic, and air pollution), and other risk factors (cigarette smoke, alcohol drinking, body size, physical activity and diet) have been observed in epidemiologic studies though the patterns are far from consistent. Challenges in inferences from the existing data are primarily due to the cross-sectional and small size of most studies to date as well as the differences in results across assay type and source of DNA. Large, prospective studies will be needed to understand whether changes in risk factors are associated with changes in DNA methylation patterns, and if changes in DNA methylation patterns are associated with changes in disease endpoints. PMID- 21636974 TI - Effects of endocrine disruptors on imprinted gene expression in the mouse embryo. AB - Environmental endocrine disruptors (EDs) are synthetic chemicals that resemble natural hormones and are known to cause epigenetic perturbations. EDs have profound effects on development and fertility. Imprinted genes had been identified as susceptible loci to environmental insults by EDs because they are functionally haploid, and because the imprints undergo epigenetic resetting between generations. To screen for possible epigenetic perturbations caused by EDs at imprinted loci, we treated pregnant mice daily between 8.5 and 12.5 days post coitum (dpc) with di-(2-ethylhexyl)-phthalate (DEHP), bisphenol A (BPA), vinclozolin (VZ), or control oil vehicle. After isolating RNA from the placenta, yolk sac, amnion, head, body, heart, liver, lung, stomach, and intestines of 13.5 dpc embryos we measured the allele-specific expression of 38 imprinted transcripts using multiplex single nucleotide primer extension (SNuPE) assays. In this representative data set we identified only a small number of transcripts that exhibited a substantial relaxation of imprinted expression with statistical significance: Slc22a18 with 10% relaxation in the embryo after BPA treatment; Rtl1as with 11 and 16% relaxation in the lung and placenta, respectively after BPA treatment; and Rtl1 with 12% relaxation in the yolk sac after DEHP treatment. Additionally, the standard deviation of allele-specificity increased in various organs after ED treatment for several transcripts including Igf2r, Rasgrf1, Usp29, Slc38a4, and Xist. Our data suggest that the maintenance of strongly biased monoallelic expression of imprinted genes is generally insensitive to EDs in the 13.5 dpc embryo and extra-embryonic organs, but is not immune to those effects. PMID- 21636975 TI - Methylation variation at IGF2 differentially methylated regions and maternal folic acid use before and during pregnancy. AB - Folic acid (FA) supplementation before and during pregnancy has been associated with decreased risk of neural tube defects although recent reports suggest it may also increase the risk of other chronic diseases. We evaluated exposure to maternal FA supplementation before and during pregnancy in relation to aberrant DNA methylation at two differentially methylated regions (DMRs) regulating Insulin-like Growth Factor 2 (IGF2) expression in infants. Aberrant methylation at these regions has been associated with IGF2 deregulation and increased susceptibility to several chronic diseases. Using a self-administered questionnaire, we assessed FA intake before and during pregnancy in 438 pregnant women. Pyrosequencing was used to measure methylation at two IGF2 DMRs in umbilical cord blood leukocytes. Mixed models were used to determine relationships between maternal FA supplementation before or during pregnancy and DNA methylation levels at birth. Average methylation at the H19 DMR was 61.2%. Compared to infants born to women reporting no FA intake before or during pregnancy, methylation levels at the H19 DMR decreased with increasing FA intake (2.8%, p=0.03, and 4.9%, p=0.04, for intake before and during pregnancy, respectively). This methylation decrease was most pronounced in male infants (p=0.01). Methylation alterations at the H19 DMR are likely an important mechanism by which FA risks and/or benefits are conferred in utero. Because stable methylation marks at DMRs regulating imprinted genes are acquired before gastrulation, they may serve as archives of early exposures with the potential to improve our understanding of developmental origins of adult disease. PMID- 21636976 TI - Timing is everything: the when and how of environmentally induced changes in the epigenome of animals. AB - Environmental influence on developmental plasticity impacts a wide diversity of animal life from insects to humans. We now understand the epigenetic basis for many of these altered phenotypes. The five environmental factors of nutrition, behavior, stress, toxins, and stochasticity work individually and in concert to affect the developing epigenome. During early embryogenesis, epigenetic marks, such as DNA methylation, are reset at specific times. Two waves of global demethylation and reestablishment of methylation frame the sensitive times for early environmental influences and will be the focus of this review. Gene transcription, translation, and post-translational modification of chromatin remodeling complexes are three mechanisms affected by developmental exposure to environmental factors. To illustrate how changes in the early environment profoundly affect these mechanisms, we provide examples throughout the animal kingdom. Herein we review the history, time points, and mechanisms of epigenetic gene-environment interaction. PMID- 21636977 TI - Human SIRT1 associates with mitotic chromatin and contributes to chromosomal condensation. AB - SIRT1 is a NAD-dependent deacetylase that participates in cellular controls of gene expression, metabolism, genomic stability and anti-aging. Here we report that SIRT1 levels rise in prometaphase leading to SIRT1 global association with mitotic chromatin until telophase. Moreover, SIRT1 contributes to chromosomal condensation by mediating chromosomal loading of histone H1 and the condensin I complex. Consistently, SIRT1 knockdown led to improper condensation and overall aberrant mitosis. Our data highlight new role for SIRT1 in maintenance of chromosome stability in mitosis and suggests how diminished SIRT1 activity during aging and tumorigenesis may lead to aneuploidy and genomic instability. PMID- 21636979 TI - Dismantling the NPC permeability barrier at the onset of mitosis. PMID- 21636978 TI - Gfer is a critical regulator of HSC proliferation. AB - Hematopoietic stem cells (HSC) are a relatively quiescent pool of cells that perform the arduous task of replacing the short-lived mature cells of the peripheral blood. While a rapid expansion of HSCs under periods of hematological stress is warranted, their enhanced proliferation during homeostasis leads to loss of function. We recently reported that in HSCs, the evolutionarily conserved growth factor erv1-like (Gfer) acts to counter jun activation domain-binding protein 1 (Jab1)-mediated nuclear export and destabilization of the cell cycle inhibitor, p27kip1, by directly binding to and sequestering the COP9 signalosome (CSN) subunit. Through this mechanism, Gfer promotes quiescence and maintains the functional integrity of HSCs. Here, we extend our study to demonstrate an association between Gfer and Ca2+/calmodulin-dependent protein kinase IV (CaMKIV) in the regulation of HSC proliferation. Highly proliferative and functionally deficient Camk4-/- HSCs possess significantly lower levels of Gfer and p27kip1. Ectopic expression of Gfer restores quiescence and elevates p27kip1 expression in Camk4-/- HSCs. These results further substantiate a critical role for Gfer in the restriction of unwarranted proliferation in HSCs through the inhibition of Jab1 and subsequent stabilization and nuclear retention of p27kip1. This Gfer-mediated pro-quiescence mechanism could be therapeutically exploited in the treatment of hematological malignancies associated with elevated Jab1 and reduced p27kip1. PMID- 21636980 TI - Disconnecting XRCC1 and DNA ligase III. AB - DNA strand break repair is essential for the prevention of multiple human diseases, particularly those which feature neuropathology. To further understand the pathogenesis of these syndromes, we recently developed animal models in which the DNA single-strand break repair (SSBR) components, XRCC1 and DNA Ligase III (LIG3), were inactivated in the developing nervous system. Although biochemical evidence suggests that inactivation of XRCC1 and LIG3 should share common biological defects, we found profound phenotypic differences between these two models, implying distinct biological roles for XRCC1 and LIG3 during DNA repair. Rather than a key role in nuclear DNA repair, we found LIG3 function was central to mitochondrial DNA maintenance. Instead, our data indicate that DNA Ligase 1 is the main DNA ligase for XRCC1-mediated DNA repair. These studies refine our understanding of DNA SSBR and the etiology of neurological disease. PMID- 21636981 TI - Time it was, and what a time it was... PMID- 21636982 TI - Beware of metaphors: chasses and orthogonality in synthetic biology. AB - Every descriptive language is not only metaphoric and interpretative, but it is also developed (or adopted) ad hoc to fulfill a certain agenda. Even the hardcore scientific languages of mathematics and physics are not entirely neutral--let alone the much softer terminology of biology. We use metaphors all the time in science and that is useful as long as they serve their purpose well and we are aware of them. But it is a serious mistake to identify uncritically the thing and the metaphors employed to bring the thing to mind. I address here two of the most sparkling metaphors brought about by synthetic biology: chassis and orthogonality -the first borrowed instance from engineering of vehicles, and the second from computer terminology (itself borrowed from mathematics). For the sake of simplicity the discussion is limited to the connotation of these two concepts in the prokaryotic realm. The power of such metaphors from describing the state of affairs in synthetic biology and for setting a vigorous research and technology schedule is analyzed. Awareness of the meta-languages and allegories that are being adopted by the SB community should help to frame other controversial concepts such as artificial cells or synthetic life. PMID- 21636983 TI - Bacterial luciferase reporters: the Swiss army knife of molecular biology. AB - Bioluminescence is a process during which light in the visible spectrum is emitted as a consequence of an enzymatic reaction catalyzed by luciferases. Luciferases have been identified mainly in marine organisms and are used for several biological purposes include camouflage, repulsion, attraction, communication and illumination. Some of the currently known luciferases have become indispensible tools in modern molecular biology and are used for diverse applications such as autoinducer-1 activity assays, promoter test assays in both prokaryotes and eukaryotes, imaging of bacterial infections in live animals, in vivo activity assays genes involved in host response and disease and monitoring of bacterial contaminations of food products. With the present review, the authors intend to give an overview on the currently used bacterial luciferase reporter systems, their methodologies and applications and compare them to other reporter systems. PMID- 21636984 TI - Proteomics in the microbial sciences. AB - Mass spectrometry based proteomics is now widely used in the microbial sciences. In conjunction with transcriptomics it has greatly enhanced the field of microbial biology and has provide microbiologists with unparalleled insights into cellular processes and functions. Proteomics allows the dynamic nature of the entire protein network to be mapped providing a deeper understanding of microbial systems, their evolution and role in disease states. This review is intended to provide an overview of mass spectrometry and its application to the field of microbial proteomics. Background is provided on the core mass analyzers, including the Orbitrap mass spectrometer, and novel fragmentation processes such as Electron Transfer Dissociation which leave post-translational modifications intact on peptide backbones allowing for their identification and localization. The review will also provide information on current key quantitative technologies and the state of the art in microbial metaproteomics. PMID- 21636985 TI - Development of a broad-host-range phage cocktail for biocontrol. AB - The aims of this study were to investigate the incidence of different resistance mechanisms to phage K in a bank of Irish Staph aureus hospital strains; and to develop a broad host-range phage cocktail with enhanced lytic activity against those strains which were previously phage resistant. A bank of 180 Staph aureus strains, which included all the sequence types currently in existence in Ireland, were tested for sensitivity to phage K. Twenty nine strains were identified, which did not permit plaque formation. The phage resistance systems in the 29 strain were investigated and it was found that restriction modification (r-m) was evident in 24, an adsorption inhibition mechanism was evident in three, while two were resistant by an unidentified mechanism. Seventeen modified derivatives of phage K were developed which could circumvent all the r-m systems. Nevertheless, six of the modified phage were considered superior in terms of their individual host ranges. These six were pooled as a cocktail with phage K, which then lysed 24 of the 29 resistant strains (97.2% of the entire staphylococcal bank). In conclusion, phage resistant systems affecting phage K are common in Staph. aureus but it is possible to significantly broaden the host-range of this phage for biocontrol applications. PMID- 21636986 TI - Transformation of the mycorrhizal fungus Laccaria bicolor using Agrobacterium tumefaciens. AB - Most boreal and temperate forest trees form a mutualistic symbiosis with soil borne fungi called ectomycorrhiza (ECM). In this association both partners benefit due to nutrient exchange at the symbiotic interface. Laccaria bicolor is the first mycorrhizal fungus with its genome sequenced thus making possible for the first time to analyze genome scale gene expression profiles of a mutualistic fungus. However, in order to be able to take full advantage of the genome sequence, reverse genetic tools are needed. Among them a high throughput transformation system is crucial. Herein we present a detailed protocol for genetic transformation of L. bicolor by means of Agrobacterium tumefaciens with emphasis on critical steps affecting the success and efficiency of the approach. PMID- 21636987 TI - Efficient E. coli expression systems for the production of recombinant beta mannanases and other bacterial extracellular enzymes. AB - Two Escherichia coli expression systems based on T7 RNA polymerase promoter (pET system) and tac promoter (pFLAG system) have been used for the production and secretion of recombinant beta-mannanases from Bacillus sp. Both E. coli OmpA signal peptide and native Bacillus signal peptide could be used efficiently for the secretion of recombinant enzymes into periplasmic space and culture media. The genes could be induced for over-expression with 0.1-1 mM isopropyl-beta-D-1 thiogalactopyranoside (IPTG) when the OD 600 of the culture broth reached 0.6 1.5. The recombinant enzymes could be harvested from whole cell lysate, perimplasmic extract, or culture broth after induction for 4-20 hours. Since the enzyme is C-terminally tagged with hexahistidine, the recombinant enzymes could be conveniently purified to apparent homogeneity by one-step immobilized-metal affinity chromatography (IMAC) using Ni-NTA resins. The characteristics of purified recombinant beta-mannanases from B. licheniformis and B. subtilis, which share 78% amino acid identity, are slightly different. These systems should be applicable for the production of various recombinant bacterial extracellular enzymes. PMID- 21636988 TI - The microalga Chlamydomonas reinhardtii as a platform for the production of human protein therapeutics. AB - Microalgae are a diverse group of eukaryotic photosynthetic microorganisms. While microalgae play a crucial role in global carbon fixation and oxygen evolution, these organisms have recently gained much attention for their potential role in biotechnological and industrial applications, such as the production of biofuels. We investigated the potential of the microalga Chlamydomonas reinhardtii to be a platform for the production of human therapeutic proteins. C. reinhardtii is a unicellular freshwater green alga that has served as a popular model alga for physiological, molecular, biochemical and genetic studies. As such, the molecular toolkit for this microorganism is highly developed, including well-established methods for genetic transformation and recombinant gene expression. We transformed the chloroplast genome of C. reinhardtii with seven unrelated genes encoding for current or potential human therapeutic proteins and found that four of these genes supported protein accumulation to levels that are sufficient for commercial production. Furthermore, the algal-produced proteins were bioactive. Thus, the microalga C. reinhardtii has the potential to be a robust platform for human therapeutic protein production. PMID- 21636989 TI - Genetic improvement of butanol tolerance in Escherichia coli by cell surface expression of fish metallothionein. AB - Cysteine-rich metallothioneins (MTs) have been reported to possess the capacity to scavenge reactive oxygen species in vitro and in vivo. Recombinant strains of Escherichia coli expressing outer membrane protein C (OmpC) fused with MTs from human, mouse and tilapia displayed the ability for such surface-localized MTs to scavenge extracellular free radicals, but the benefits of the possible applications of this capacity have not yet been demonstrated. Because the intrinsic butanol tolerance of microbes has become an impediment for biological butanol production, we examined whether surface-displayed MTs could contribute to butanol tolerance. The results show that strains expressing OmpC-MT fusion proteins had higher butanol tolerance than strains with cytoplasmically expressed MTs. Furthermore, the OmpC-tilapia MT fusion protein enhanced butanol tolerance more strongly than other recombinant constructs. Although the enhanced level of tolerance was not as high as that provided by OmpC-tilapia MT, over-expression of OmpC was also found to contribute to butanol tolerance. These results suggest that free-radical scavenging by MT and OmpC-related osmoregulation enhance butanol tolerance. Our results shed new light on methods for engineering bacteria with higher butanol tolerance. PMID- 21636990 TI - Parenteral vaccination of mammalian livestock with Newcastle disease virus-based vector vaccines offers optimal efficacy and safety. AB - Newcastle disease virus (NDV) is an avian virus that is being evaluated as a vaccine vector for the delivery of foreign genes in mammals. The use of NDV as a vaccine vector in these species offers two major advantages. First, NDV is highly attenuated in mammals, rendering its use inherently safe. Second, mammals lack pre-existing NDV immunity, which minimizes the risk of vaccination failure. NDV vector vaccines are generally administered to mammals via the respiratory route. We recently showed that intramuscular vaccination with NDV-based Rift Valley fever virus (RVFV) vaccines provides complete protection in mice and induces neutralizing antibodies in sheep and cattle, the main target species of RVFV. Here, we discuss the use of NDV as a vaccine vector for applications in mammalian livestock with an emphasis on the vaccination route. We also report the results of novel experiments that underscore our notion that vaccination via a parenteral route is more effective than immunization via the respiratory route. PMID- 21636991 TI - Escherichia coli K5 heparosan fermentation and improvement by genetic engineering. AB - N-acetyl heparosan is the precursor for the biosynthesis of the important anticoagulant drug heparin. The E. coli K5 capsular heparosan polysaccharide provides a promising precursor for in vitro chemoenzymatic production of bioengineered heparin. This article explores the improvements of heparosan production for bioengineered heparin by fermentation process engineering and genetic engineering. PMID- 21636992 TI - Stem cell research in the Emerald Isle. AB - Fiscal policy, governance, education and a more internationally integrative strategic policy significantly assists Ireland's competitiveness in stem cell R&D sector. PMID- 21636993 TI - Introducing a new bioengineered bug: Methylobacterium extorquens tuned as a microbial bioplastic factory. AB - Discussion on and use of methanol as chemical feedstock and as alternative fuel has gained momentum during the past years. Consequently, microorganism and product design based on ''methylotrophism'' is in vogue as reflected by increasing research and development activities in methanol-related areas. A recent example of microorganism and product development is the use of recombinant Methylobacterium extorquens ATCC 55366 strains in the production of second generation biopolyesters. Feeding n-alkenoic acids in addition to methanol yielded functionalized polyhydroxyalkanoates (PHAs) and uncovered how M. extorquens copes with fatty acids. While some parts of the degradation pathway remain unclear, possible metabolic routes are suggested that may explain the significant loss of double bonds prior to polymerization of PHA precursors and occurrence of odd-numbered monomers derived from even-numbered n-alkenoic acids. In addition, microbial discoloration upon fatty acid feeding is discussed and future directions for further genetic modification of M. extorquens are provided. PMID- 21636994 TI - Engineering lactic acid bacteria for increased industrial functionality. AB - Based on their spoilage-preventing and flavor-contributing characteristics, lactic acid bacteria (LAB) are employed as starter cultures for the fermentation of foods and feeds. In addition, several specific LAB strains are marketed on basis of their beneficial effects on the consumer's health, representing an explosively growing market for the products containing these so-called probiotics. Due to this extensive industrial use there is a strong interest in unraveling the molecular mechanisms involved in industrial robustness, cognate stress resistance, and health-promoting phenotypes of these LAB that may vary drastically between different starter and probiotic strains currently marketed. This review describes some of the post-genomic tools developed, as well as their employment for the identification of bacterial effector molecules involved in the aforementioned industrially relevant phenotypes. Furthermore, it addresses possible strategies to exploit such knowledge into the rational design of LAB strains with increased industrial functionality. PMID- 21636995 TI - Comparative modelling of LysB from the mycobacterial bacteriophage Ardmore. AB - Given their potential as specific and natural biocontrol agents, bacteriophages and their associated proteins have become the focus of renewed attention over the last decade. The aim of this study was to use a comparative modelling approach to generate a predicted 3D structure for LysB; a 332 amino acid lipolytic enzyme encoded by the mycobacteriophage Ardmore. The GXSXG pentapeptide, characteristic of lipolytic enzymes, was located at amino acid position 166-170. The three absolutely conserved residues among mycobacteriophage LysB proteins were also identified in Ardmore LysB as Ser-168, Gly-203 and Pro-205. CATH analysis of Ardmore LysB revealed a mainly Beta classification, Beta Barrel architecture and a topology similar to maltoporin. This is unlike the alpha/beta hydrolase structure reported for the D29 LysB protein and, in fact appears in only 3 other sequenced LysB homologues to date. A search for conserved motifs within the amino acid sequence of LysB revealed the presence of both a cutinase motif and a PE-PPE motif. This study presents an in silico 3D predictive model of Ardmore lysin and confirms the high diversity of mycobacteriophages LysB proteins both at the sequence (2D) and structural (predicted 3D) levels. PMID- 21636996 TI - LysGH15 reduces the inflammation caused by lethal methicillin-resistant Staphylococcus aureus infection in mice. AB - The endolysin LysGH15, derived from staphylococcal phage GH15, has a wide lytic spectrum and strong lytic activity against Staphylococcus aureus, especially methicillin-resistant S. aureus (MRSA), in vitro and in vivo. Here, the ability of lethal MRSA to induce mRNA levels of interleukin-6 (IL-6), interleukin-4 (IL 4), and interferon-gamma (IFN-gamma) in spleen tissues of mice was studied. A large number of bacteria were detected in spleens. The bacteria caused elevated expression levels of these three cytokines. Administration of LysGH15 significantly reduced the number of bacteria and the levels of IL-6, IL-4, and IFN-gamma mRNA in spleen cells compared with those in untreated mice at 24 h (P < 0.05). LysGH15 can eliminate a large number of bacteria and effectively alleviate inflammation induced by infection with lethal MRSA. PMID- 21636997 TI - Filamentous fungi in good shape: microparticles for tailor-made fungal morphology and enhanced enzyme production. AB - Filamentous fungi such as Aspergillus niger are important biocatalysts for industrial production of various enzymes as well as organic acids or antibiotics. In suspended culture these microorganisms exhibit a complex morphology which typically has a strong influence on their production properties. In this regard, we have recently shown that the addition of inorganic micro particles to the culture medium is a straightforward and elegant approach to precisely tame fungal morphology. For A. niger a full range of morphological forms from pellets with different diameters to free mycelium could be adjusted by supplementation with talc powder. Aluminium oxide particles similarly affected morphology, showing that this effect is largely independent of the chemical particle composition. Exemplified for different recombinant A. niger strains enzyme production could be strongly enhanced by the addition of microparticles. This was demonstrated for the production of fructofuranosidase, an important high-value biocatalyst for pre biotic fructo-oligosaccharides, by recombinant A. niger. In a microparticle enhanced fed-batch process, a highly productive mycelium could be achieved. The enzyme titre of 2800 U/mL finally reached was more then tenfold higher then that of any other process reported so far. Here we provide additional insights into the novel production process. This includes the confirmation of the highly selective production of the target enzyme fructofuranosidase using MALDI-TOF MS analysis. Moreover, we show that the obtained enzyme suspension can be efficiently used with minimal pre-treatment for the biosynthesis of short chain fructooligosaccharides of the inulin type, such as 1-kestose and 1-nystose, prebiotics with substantial commercial interest. In particular, these compounds are highly attractive for human consumption, since they have been shown to reduce the risk of colon cancer. In summary, the use of microparticles opens a new avenue of engineering fungal morphology into the desired form for specific production processes. PMID- 21636998 TI - Molecular responses of Escherichia coli caused by heat stress and recombinant protein production during temperature induction. AB - In a recent review, we discussed the extensively used temperature-inducible expression system, based on the pL and/or pR phage lambda promoters that are finely regulated by the thermo-labile cI857 repressor. In this system, an increase in temperature induces the heterologous protein production and activates the heat shock response, as well as the stringent and SOS responses. The same responses are activated just by the overproduction of recombinant protein. All such responses result in a metabolic burden to the cells, a decrease in the specific growth rate, and alterations in the central carbon metabolism. Altogether, these effects can alter the quantity and quality of the produced foreign protein. Here, we compare and discuss the transcription of selected genes, and the concomitant synthesis of heat-shock proteins (hsp) soon after thermal induction, in relation to the responses that occur in other expression systems that also trigger the heat-shock response. PMID- 21636999 TI - Characteristics of artificial forisomes from plants and yeast. AB - Forisomes are protein bodies found exclusively in the phloem of the Fabaceae (legumes). In response to wounding, the influx of Ca ( 2+) induces a conformational change from a condensed to a dispersed state which plugs the sieve tubes and prevents the loss of photoassimilates. This reversible, ATP-independent reaction can be replicated with purified forisomes in vitro by adding divalent cations or electrically inducing changes in pH, making forisomes ideal components of technical devices. Although native forisomes comprise several subunits, we recently showed that functional homomeric forisomes with distinct properties can be expressed in plants and yeast, providing an abundant supply of forisomes with tailored properties. Forisome subunits MtSEO-F1 and MtSEO-F4 can each assemble into homomeric artificial forisomes, which indicates functional redundancy. However, we provide further evidence that both proteins are subunits of the native heteromeric forisome body in planta. We also show that the properties of artificial forisomes can be modified by immobilization, which is a prerequisite for their incorporation into technical devices. PMID- 21637000 TI - Is it possible to produce succinic acid at a low pH? AB - Bio-based succinate is still a matter of special emphasis in biotechnology and adjacent research areas. The vast majority of natural and engineered producers are bacterial strains that accumulate succinate under anaerobic conditions. Recently, we succeeded in obtaining an aerobic yeast strain capable of producing succinic acid at low pH. Herein, we discuss some difficulties and advantages of microbial pathways producing "succinic acid" rather than "succinate." It was concluded that the peculiar properties of the constructed yeast strain could be clarified in view of a distorted energy balance. There is evidence that in an acidic environment, the majority of the cellular energy available as ATP will be spent for proton and anion efflux. The decreased ATP:ADP ratio could essentially reduce the growth rate or even completely inhibit growth. In the same way, the preference of this elaborated strain for certain carbon sources could be explained in terms of energy balance. Nevertheless, the opportunity to exclude alkali and mineral acid waste from microbial succinate production seems environmentally friendly and cost-effective. PMID- 21637001 TI - Oxidative versus reductive succinic acid production in the yeast Saccharomyces cerevisiae. AB - Bio-based succinic acid is receiving increasing attention, as it could provide a cost-effective, ecologically sustainable alternative to the current petrochemical production process, thus promising a significantly higher market potential. The yeast Saccharomyces cerevisiae is a robust and well-established industrial production organism exhibiting an extraordinarily high acid- and osmotolerance. These features in conjunction with the sophisticated toolbox for genetic engineering make it particularly suitable for succinic acid production. The high tolerance towards acidity is a major advantage over previously established bacterial succinic acid production hosts, since it makes the use of neutralisation salts dispensable and thus enormously facilitates the downstream process. By constructing yeast strains capable of producing significant amounts of succinic acid, we have recently established S. cerevisiae as a promising host for succinic acid production. Our metabolic engineering strategy relied on the implementation of an oxidative production route using the glyoxylate cycle. We here discuss theoretical and practical aspects of oxidative and reductive succinic acid production routes in S. cerevisiae. PMID- 21637002 TI - Caspase and nm23: apoptosis genes linked to the antibacterial response of the Chinese mitten crab. AB - Apoptosis is a central regulatory feature of the immune system, and the most common form of death among immunological cells. However, the function of apoptosis, within the innate immune system of invertebrates, remains largely unknown. For this reason, we investigated the immune functionality of two apoptosis genes, caspase and nm23, in the Chinese mitten crab (Eriocheir sinensis), a commercially important and disease vulnerable aquaculture species. The capase and nm23 transcripts were detected in all examined crab tissue; furthermore, the expression is immuno-responsive and may be linked to immune functionality in crabs. PMID- 21637003 TI - Modulation of RNA splicing as a potential treatment for cancer. AB - Close to 90% of human genes are transcribed into pre-mRNA that undergoes alternative splicing, producing multiple mRNAs and proteins from single genes. This process is largely responsible for human proteome diversity, and about half of genetic disease-causing mutations affect splicing. Splice-switching oligonucleotides (SSOs) comprise an emerging class of antisense therapeutics that modify gene expression by directing pre-mRNA splice site usage. Bauman et al. investigated an SSO that up-regulated the expression of an anti-cancer splice variant while simultaneously eliminating an over-expressed cancer-causing splice variant. This was accomplished by targeting pre-mRNA of the apoptotic regulator Bcl-x, which is alternatively spliced to express anti- and pro-apoptotic splice variants Bcl-xL and Bcl-xS, respectively. High expression of Bcl-xL is a hallmark of many cancers and is considered a general mechanism used by cancer cells to evade apoptosis. Redirection of Bcl-x pre-mRNA splicing from Bcl-xL to -xS by SSO induced apoptotic and chemosensitizing effects in various cancer cell lines. Importantly, the paper shows that delivery of Bcl-x SSO using a lipid nanoparticle redirected Bcl-x splicing and reduced tumor burden in melanoma lung metastases. This was the first demonstration of SSO efficacy in tumors in vivo. SSOs are not limited to be solely potential anti-cancer drugs. SSOs were first applied to repair aberrant splicing in thalassemia, a genetic disease, they have been used to create novel proteins (e.g., ?7TNFR1), and they have recently progressed to clinical trials for patients with Duchenne muscular dystrophy. PMID- 21637004 TI - Engineered cyanobacteria: teaching an old bug new tricks. AB - Cyanobacteria have played an important role in the development of the Earth and have long been studied as model organisms for photosynthesis and the circadian rhythm. Recent developments have led to increased interest in the use of engineered cyanobacteria for the production of protein and chemical products. This review highlights the genetic tools and strategies for manipulation of cyanobacteria as well as previous accomplishments in the development of engineered cyanobacteria for applied use. Particular attention is given to the engineering of cyanobacteria for biofuel production, including both hydrocarbon and hydrogen fuels. Genetic engineering efforts to enhance cyanobacterial fitness are reviewed with an emphasis on physiological improvements for large-scale production. Lastly, a future outlook on engineered cyanobacteria is presented, highlighting the future areas of focus and technical challenges in this field. With the uncertainty of future energy security, it is an exciting time in applied cyanobacterial research, but we must take the time to learn from these past accomplishments before we can capitalize on the potential of these photosynthetic microorganisms. PMID- 21637005 TI - Cell immobilization studies using a cellulose-binding domain fused to PrtP in Lactococcus lactis. AB - The cellulose-binding domain (CBD) of XylA was fused with PrtP, NisP and AcmA anchors derived from Lactococcus lactis under P45 promoter and Usp45 secretion signal. The fusion construct with the anchor PrtP (334 aa) was shown to exhibit the most efficient immobilization. The CBD-PrtP construct on the other hand was not efficiently attached to the cell wall and as such was found mainly in the supernatant. Results also showed that expression of the CBD-NisP anchor fusion led to a similar result. This raised the question if more efficient binding of the anchor to the cell wall by sortase could enhance the efficiency of cell immobilization to the cellulosic material. However, expressing sortase with the CBD-PrtP fusion did not improve the immobilization of the cells to cellulose. PMID- 21637006 TI - Vaccine and adjuvant design for emerging viruses: mutations, deletions, segments and signaling. AB - Vaccination is currently the most effective strategy to medically control viral diseases. However, developing vaccines is a long and expensive process, and traditional methods, such as attenuating wild-type viruses by serial passage, may not be suitable for all viruses and may lead to vaccine safety considerations, particularly in the case of the vaccination of particular patient groups, such as the immunocompromised and the elderly. In particular, developing vaccines against emerging viral pathogens adds a further level of complexity, as they may only be administered to small groups of people or only in response to a specific event or threat, limiting our ability to study and evaluate responses. In this commentary, we discuss how novel techniques may be used to engineer a new generation of vaccine candidates as we move toward a more targeted vaccine design strategy, driven by our understanding of the mechanisms of viral pathogenesis, attenuation and the signaling events which are required to develop a lasting, protective immunity. We will also briefly discuss the potential future role of vaccine adjuvants, which could be used to bridge the gap between vaccine safety, and lasting immunity from a single vaccination. PMID- 21637007 TI - E. coli filament formation induced by heterologous S-layer expression. AB - Escherichia coli is a rod-shaped intestinal bacterium which has a size of 1.1-1.5 um x 2.0-6.0 um. The fast cell division process and the uncomplicated living conditions have turned E. coli into a widely used host in genetic engineering and into one of the best studied microorganisms of all. We used E. coli BL21(DE3) as host for heterologous expression of S-layer proteins of Lysinibacillus sphaericus JG-A12 in order to enable a fast and high efficient protein production. The S layer expression induced in E. coli an unusual elongation of the cells, thus producing filaments of > 100 um in length. In the stationary growth phase, E. coli filaments develop tube-like structures that contain E. coli single cells. Fluorescence microscopic analyses of S-layer expressing E. coli cells that were stained with membrane stain FM ((r)) 5-95 verify the membrane origin of the tubes. Analyses of DAPI stained GFP-S-layer expressing E. coli support the assumption of a disordered cell division that is induced by the huge amount of recombinant S-layer proteins. However, the underlying mechanism is still not characterized in detail. These results describe the occurrence of a novel stable cell form of E. coli as a result of a disordered cell division process. PMID- 21637008 TI - Recombinant adeno-associated virus expressing a modified transcription factor triggered by the hepatitis C viral protease NS3/4a. AB - Many viruses, including hepatitis C virus (HCV), are major threats to public health, but few treatment options are available. Systemic administration of a combination of interferon-a and ribavirin is the only approved treatment for HCV. However, half of all patients are not cured by such treatment and a wide spectrum of systemic side-effects limits its effectiveness. I developed a gene therapy approach with three goals: 1) the restoration of local interferon secretion in cells infected with HCV, 2) no secretion of interferon in normal cells not infected with the virus, and 3) suppression of the emergence of resistant viral strains. A recombinant transcription factor was constructed, the intracellular localization of which is controlled by a viral protease to stimulate focal interferon secretion at sites of infection. A recombinant adenovirus associated virus expressing the transcription factor based on the described strategy inhibited HCV replication effectively in a HCV in vitro culture system. PMID- 21637009 TI - Identification and characterization of an essential gene in Listeria monocytogenes using an inducible gene expression system. AB - The Listeria monocytogenes gene lmo1594 is a homolog of the Bacillus subtilis cell division gene ezrA. EzrA is a negative regulator of FtsZ ring formation, which is required for efficient cell division as it regulates the frequency and position of Z-rings in the cell and prevents aberrant polar cell division. Previously identified as a putative high pressure (HP) resistance mechanism; conferring enhanced barotolerance when heterologously expressed against an Escherichia coli background; the aim of the current study was to investigate whether lmo1594 plays a role in listerial barotolerance. When the creation of a deletion mutant proved unsuccessful, the role of lmo1594 was addressed by creating a conditional knockout mutant which demonstrated that the gene is in fact essential for cell survival and growth in L. monocytogenes. In order to investigate the effect of lmo1594 on barotolerance, the gene was over-expressed. The over-expression of lmo1594 increased survival levels in L. monocytogenes treated at 300 MPa, but survival levels similar to those of the wild-type strain were observed when treated at a higher pressure (>=400 MPa). In conclusion, this study reveals for the first time that lmo1594 is absolutely essential for listerial cell growth and survival, and also plays an important role in listerial barotolerance. PMID- 21637010 TI - Exploiting recombinant antibodies in point-of-care (POC) diagnostics: the combinatorial advantage. AB - Antibodies are ubiquitously deployed on in vitro diagnostic (IVD) platforms for detecting a panoply of analytes indicative of environmental and food contamination, residue adulteration and both veterinary and medical diagnostics. In the clinical realm, rapid and accurate determination of disease status is paramount. The significance of immunodiagnostic performance cannot be overemphasized and in many cases reliable diagnosis informs medical intervention which can mean the difference between patient recovery and demise. Cardiovascular disease (CVD) is the single biggest cause of adult mortality in the western world and principal burden on the healthcare services. Although the troponin (Tn) family, in particular troponin I (TnI), are regarded as the gold standard for diagnosis of myocardial damage, over the last decade much research has focused on the identification of alternative cardiac biomarker molecules that can either supplant or complement TnI metrics to add value to cardiac risk stratification criteria. In particular, markers that appear earlier than TnI in the pathophyisiology of cardiac disease are highly sought after. The subject of this addendum represents part of a broader challenge to deliver novel rapid point-of care (POC) diagnostics to provide a chip-based multi-plexed platform for more comprehensive profiling of cardiac status with additive diagnostic and prognostic value. Specifically, it outlines proof-of-concept direct myeloperoxidase (MPO) detection, demonstrates the benefits of using recombinant antibodies in POC diagnostics and describes optimized strategies for generation of superior candidate antibody panels. PMID- 21637012 TI - Mind-bending infections. PMID- 21637011 TI - A new diet for yeast to improve biofuel production. AB - In 2010, our group announced the discovery of two cellodextrin transporter families from the cellulolytic fungus, Neurospora crassa. Furthermore, we demonstrated the utility of these transporters in the production of lignocellulosic biofuels. This discovery was made possible by a decision to systematically study cell wall degradation by N. crassa. The identified transport pathway has opened up a new way of thinking about microbial fermentation of hexoses as well as pentoses derived from plant cell walls. Integrating this pathway with the endogenous metabolism and signaling networks of S. cerevisiae is now a major goal of our group. PMID- 21637013 TI - A review of Dientamoeba fragilis carriage in humans: several reasons why this organism should be considered in the diagnosis of gastrointestinal illness. AB - Dientamoeba fragilis is a protozoan that inhabits the human gut. It is approximately 100 years since Dientamoeba's discovery and first description when it was described as a rare and harmless commensal. Since then it has struggled to gain recognition as a pathogen despite the evidence supporting its pathogenicity. Dientamoeba remains neglected, probably due to the misconceptions that it is uncommon and non-pathogenic. Usually, carriage of Dientamoeba is associated with symptoms such as abdominal pain and diarrhea. Moreover, antimicrobial therapy followed by resolution of symptoms coincides with the eradication of Dientamoeba. This manuscript reviews the scientific literature relating to Dientamoeba's prevalence and pathogenicity. While much of the evidence supporting its pathogenicity is only circumstantial, it is apparent that most researchers agree that Dientamoeba is pathogenic. Therefore, in symptomatic patients who harbor Dientamoeba and no other pathogen, Dientamoeba should be considered as the etiological agent and treated as such. PMID- 21637014 TI - Autotransporters and virulence of enteroaggregative E. coli. AB - Enteroaggregative Escherichia coli (EAEC) is an emerging pathogen associated with acute and persistent diarrhea in children and adults. EAEC strains are very heterogeneous and the pathogenesis of EAEC diarrhea is complex and not completely understood. Studies have suggested three major features of EAEC pathogenesis: abundant adherence to the intestinal mucosa, elaboration of enterotoxins and cytotoxins, and induction of mucosal inflammation. Here, we discuss the role of the virulence factors involved in these three major features, focusing in the EAEC adhesion including fimbrial and afimbrial factors, EAEC toxins and autotransporter proteins, such as Pet (plasmid encoded toxin) and Pic (protein involved in colonization); both proteins play a role in two EAEC pathogenic features: cytotoxicity and mucosal colonization, including the bacterium-mucus biofilm. Finally we discuss relevant factors involved in the inflammatory process induce by EAEC, such as flagellin, fimbria and regulator factors (AggR). Interestingly, all these factors are not present in all EAEC strain, contributing to EAEC heterogeneity. PMID- 21637015 TI - Human intestinal microbiota: characterization of a simplified and stable gnotobiotic rat model. AB - The study of host microbe interactions is hampered by the complexity and inter individual variability of the human gut microbiota. Therefore, a simplified human intestinal microbiota (SIHUMI) consisting of seven bacterial species was introduced into germfree rats. Species selection was based on numerical importance and fermentative abilities in the human gut. Association of the rats with the SIHUMI (Anaerostipes caccae, Bacteroides thetaiotaomicron, Bifidobacterium longum, Blautia producta, Clostridium ramosum, Escherichia coli and Lactobacillus plantarum) resulted in increased faecal concentrations of short chain fatty acids compared to germfree animals. Since the faecal butyrate concentration was low (0.9 +/- 0.5 umol/g dry matter) the SIHUMI was complemented with Clostridium butyricum. This extended bacterial community (SIHUMIx) led to an increased faecal butyrate concentration of 1.5 +/- 0.7 umol/g dry matter. Besides forming SCFA, the SIHUMIx was capable of degrading mucins, beta-aspartylglycine and bilirubin. These features are characteristic of conventional animals but not observed in germfree animals. Dietary interventions with modifications in fibre and fat content led to changes in the proportion of community members. The relative increase of one member of this community in response to a high-fat diet reflects the situation reported for obese mice and human subjects. The strength of the model communities is their remarkable stability over time and their easy transfer to the offspring. PMID- 21637016 TI - Determination of spatial and temporal colonization of enteropathogenic E. coli and enterohemorrhagic E. coli in mice using bioluminescent in vivo imaging. AB - Infectious diarrhea is a major contributor of child morbidity and mortality in developing nations. Murine models to study the pathogenesis of infectious diarrhea caused by organisms such as enteropathogenic E. coli (EPEC) and enterohemorrhagic E. coli (EHEC) are not fully characterized. More emphasis has been placed on infection of mice with the murine specific pathogen Citrobacter rodentium. While these three organisms are genetically related they are not identical. Our goal was to better characterize the murine model of EPEC and EHEC infection by using bioluminescent bacteria to determine temporal and spatial colonization of these two human pathogens. EPEC and EHEC were transformed with a bacterial luciferase expression plasmid containing the constitutive OmpC promoter. C57BL/6 mice were orally inoculated with bioluminescent EPEC or EHEC and bacterial localization in the intestine was monitored ex vivo and in vivo by IVIS. At 3 days after infection, EPEC, EHEC and Citrobacter rodentium were all localized in the cecum and colon. EPEC colonization peaked at day 2-3 and was undetectable by day 7. The bioluminescent EPEC adheres to the cecum and colon of the mouse intestine. However, when EPEC infected mice were administered xylazine/ketamine for in vivo live imaging, the EPEC persisted at high densities for up to 31 days. This is the first report of a bioluminescent imaging of luciferase expressing EPEC in a mouse model. PMID- 21637017 TI - Life at the margins: modulation of attachment proteins in Helicobacter pylori. AB - Helicobacter pylori is the primary cause of peptic ulcer disease and is estimated to account for about 60% of all cases of gastric cancer, the second most common cause of cancer death worldwide. Among the H. pylori virulence factors associated with disease, in addition to the well-known cag pathogenicity island, is the BabA adhesin, an outer membrane protein that binds with high affinity to fucosylated glycans on the gastric epithelium, such as Lewis B (Le(b)) and related terminal fucose residues found on the blood group O (H antigen), A and B antigens. BabA mediated attachment to the gastric mucosa promotes chronic inflammation and gastric pathology, which from the bacterial perspective carries both risks and benefits. We recently described modulation in expression of BabA and related outer membrane proteins that occurs during colonization of experimental animals. Here we put these findings into a broader context, and speculate on their implications for the host-pathogen relationship. PMID- 21637018 TI - Paneth cell function--implications in pediatric Crohn disease. AB - Defects in the intestinal barrier play a central role in disease pathogenesis. Recently we have demonstrated that children with ileal Crohn's disease show a reduced expression of small intestinal HD-5 at the age of onset suggesting that a compromised mucosal barrier function might be a key factor in the early disease pathogenesis. We also identified a disturbance of the Wnt signaling transcription factor TCF-4 as a major mechanism for this deficiency in children which might result in a compromised innate immune function of small intestinal Paneth cells via defensin secretion. Here we provide a summary on our recent findings and discuss the data in more detail especially focusing on the role of Paneth cell differentiation and function in the pathogenesis of pediatric ileal Crohn's disease. PMID- 21637019 TI - Distance burning: how gut microbes promote extraintestinal cancers. AB - Gut microbes play a major role in carcinogenesis of the gastrointestinal tract. We and others have shown in mouse models that colonic bacteria also influence the development of extraintestinal cancers including hepatocellular and mammary carcinomas. Microbes such as Helicobacter hepaticus invoke a proinflammatory microenvironment in the lower bowel that may extend to distant organs, often in the absence of histologically evident inflammation. Innate immunity plays a crucial role in the promotion of liver cancer and other systemic diseases by gut microbes. Additional mechanisms include type 1 adaptive immunity, altered metabolism, and oxidative stress. Emerging links between host genetics, gut microbes, inflammatory bowel disease and colorectal cancer also may prove useful for the correlation of specific bacterial populations with extraintestinal neoplasms. Interruption of deleterious host-microbe networks through judicious use of antibiotics and targeted molecular therapies may help reduce the incidence of liver, breast, and other human cancers. PMID- 21637020 TI - A breathtaking feat: to compete with the gut microbiota, Salmonella drives its host to provide a respiratory electron acceptor. AB - Salmonella is a common cause of food poisoning. However, after ingestion the pathogen has to compete with resident microbes that already occupy the intestinal lumen (microbiota), which poses a challenge for Salmonella to successfully colonize this niche. Recent data show that Salmonella elicits help from the host immune response to beat the competition. After arriving in the intestine, Salmonella elicits acute intestinal inflammation. The respiratory burst of neutrophils that transmigrate into the intestinal lumen during inflammation oxidizes endogenous sulfur compounds to generate a respiratory electron acceptor, tetrathionate. As a result, Salmonella can use tetrathionate respiration to outgrow the fermenting microbiota in the anaerobic environment of the gut, which promotes transmission of the pathogen. This principle might be used by other gut microbes and contribute to changes in the microbiota composition observed during inflammation. PMID- 21637021 TI - Activation of type I IFN signaling by NOD1 mediates mucosal host defense against Helicobacter pylori infection. AB - Infection of gastric epithelial cells with Helicobacter pylori (H. pylori) induces a complex array of host protective immune responses. The best known are the adaptive T helper type 1 and type 17 responses that are induced in the gastric lamina propria by antigen-presenting cells via presentation of H. pylori antigens to CD4(+) T cells. Recently, it has become apparent that innate immune responses are also induced by H.pylori infection, both in epithelial cells and in underlying antigen-presenting cells. One important component of these innate responses involves the activity of NOD1, an intra-cellular sensor of peptides derived from the peptidoglycan component of the bacterial cell wall. In this review, we discuss our recent work showing that the signaling pathway utilized by NOD1 results in the generation of type I interferon and that this cytokine mediates both chemokine and cytokine responses that regulate the severity of gastric H. pylori infection. PMID- 21637022 TI - Of cabbages and kings. PMID- 21637023 TI - Mechanistic insight into polysaccharide use within the intestinal microbiota. AB - It is becoming increasingly clear that diet is one of the major factors that drives the function and composition of the intestinal microbiota. The diet of humans is highly diverse when considering different populations or even a single individual over a relatively short period of time. However, we are just beginning to understand the mechanisms that connect dietary change to intestinal microbiota dynamics. The community of microbes within our distal digestive tract influences numerous aspects of our biology, and aberrant shifts in its composition appear to be associated with several diseases. It is, therefore, necessary to understand how our behaviour and environmental factors, such as changes in diet, impact our intestinal residents. Here we look to recent work to highlight some of the major questions on the horizon for understanding the key role that the Bacteroidetes play in the commerce of dietary polysaccharides within the intestine. PMID- 21637024 TI - Cyclopropane fatty acid synthase mutants of probiotic human-derived Lactobacillus reuteri are defective in TNF inhibition. AB - Although commensal microbes have been shown to modulate host immune responses, many of the bacterial factors that mediate immune regulation remain unidentified. Select strains of human-derived Lactobacillus reuteri synthesize immunomodulins that potently inhibit production of the inflammatory cytokine TNF. In this study, genetic and genomic approaches were used to identify and investigate L. reuteri genes required or human TNF immunomodulatory activity. Analysis of membrane fatty acids from multiple L. reuteri strains cultured in MRS medium showed that only TNF inhibitory strains produced the cyclopropane fatty acid (CFA) lactobacillic acid. The enzyme cyclopropane fatty acid synthase is required for synthesis of CFAs such as lactobacillic acid, therefore the cfa gene was inactivated and supernatants from the cfa mutant strain were assayed for TNF inhibitory activity. We found that supernatants from the wild-type strain, but not the cfa mutant, suppressed TNF production by activated THP-1 human monocytoid cells Although this suggested a direct role for lactobacillic acid in immunomodulation, purified lactobacillic acid did not suppress TNF at physiologically relevant concentrations. We further analyzed TNF inhibitory and TNF non-inhibitory strains under different growth conditions and found that lactobacillic acid production did not correlate with TNF inhibition. These results indicate that cfa indirectly contributed to L. reuter immunomodulatory activity and suggest that other mechanisms, such as decreased membrane fluidity or altered expression of immunomodulins, result in the loss of TNF inhibitory activity. By increasing our understanding of immunomodulation by probiotic species, beneficial microbes can be rationally selected to alleviate intestinal inflammation. PMID- 21637025 TI - Cesarean delivery is associated with celiac disease but not inflammatory bowel disease in children. AB - The postnatal period represents a particularly dynamic phase in the establishment of the host-microbial homeostasis. The sterile protected intestinal mucosa of the fetus becomes exposed to and subsequently colonized by a complex and diverse bacterial community. Both, the exposure to microbial ligands and the bacterial colonization have been described to differ between neonates born vaginally or by cesarean delivery. These differences might influence the development of the mucosal immune system, the establishment of a stable intestinal host-microbial homeostasis, and ultimately contribute to the risk to acquire immune mediated diseases later in life. Indeed, an increased risk for atopic diseases such as allergic rhinitis and asthma was reported in children born by cesarean delivery. Our recent study described an association between cesarean delivery and celiac disease. Here we summarize the available information on postnatal microbial colonization and the influence of the mode of delivery on flora composition and host microbial homeostasis. We discuss possible consequences of the mode of delivery on epithelial barrier function and the establishment of the mucosal immune system and speculate on functional links between flora alterations and the development of inappropriate host immune responses that may contribute to enteric inflammatory diseases. PMID- 21637027 TI - A comparative analysis of the effect of antibiotic treatment and enteric infection on intestinal homeostasis. AB - The intestinal metabolome is a rich collection of molecules with specialized functions and important physiological effects. Many insults such as enteric infection and microbiota disruption by antibiotics can have profound effects in the metabolic homeostasis of the gut. We have recently shown that Salmonella infection and antibiotic treatment of mice drastically alter the intestinal metabolome. Particularly, host hormone metabolism was significantly altered by both insults. Infection resulted in a net increase in the production of both steroids and eicosanoids, whereas antibiotic treatment seemed to reduce the production of these hormones. Our results suggest that both intestinal pathogens and commensals affect common metabolic functions and that this phenomenon may have implications for the interactions between microbes and their hosts. PMID- 21637028 TI - Flexible cytokine production by macrophages and T cells in response to probiotic bacteria: a possible mechanism by which probiotics exert multifunctional immune regulatory activities. AB - Probiotics have been reported to be efficacious against cancers, infections, allergies, inflammatory bowel diseases and autoimmune diseases, and it is important to explain how such multifunctional activities are realized. Lactobacillus casei Shirota (LcS) is one of these multifunctional probiotics, and its ability to augment the host immune system has been extensively examined. We have shown that the cell wall structure of this probiotic strain is responsible for potently inducing IL-12 production. In addition, we have recently found that LcS differentially controls the inflammatory cytokine responses of macrophages and T cells in either Peyer's patches or the spleen. Other studies revealed that LcS-induced IL-12 production by macrophages is modified when other bacteria or their cell components are simultaneously present. These findings can provide a theoretical basis for understanding the multifunctional activities of specific probiotics. PMID- 21637029 TI - The endoplasmic reticulum stress response chaperone: Gp96, a host receptor for Crohn disease-associated adherent-invasive Escherichia coli. AB - Ileal lesions of Crohn disease (CD) patients are abnormally colonized by adherent invasive Escherichia coli (AIEC) producing outer membrane vesicles (OMVs) that contribute to the bacterial invasion process. In addition, endoplasmic stress was recently reported to have a key role in CD patients after the discovery, in CD patients, of several single nucleotide polymorphisms within gene encoding key component of the ER stress response and of increased expression of ER-localised stress response proteins. We recently demonstrated that the ER-stress response chaperone Gp96 is strongly expressed on the apical surface of ileal epithelial cells in CD patients and acts as a host cell receptor for OMVs, promoting AIEC invasion. In this addendum, we provide a synopsis on current data concerning AIEC virulence, summarize our recent findings and also discuss, in more detail, unresolved issues and the potential implications of our findings. PMID- 21637030 TI - Is microencapsulation the future of probiotic preparations? The increased efficacy of gastro-protected probiotics. AB - In a recent publication we assessed the kinetics of intestinal colonization by microencapsulated probiotic bacteria in comparison with the same strains given in an uncoated form. It's well known, in fact, that microencapsulation of probiotics with specific materials is able to confer a significant resistance to gastric juice, thus protecting the cells during the gastric and duodenal transit and enhancing the probiotic efficacy of any supplementation. In any case, this was the first study reporting the fecal amounts of probiotics administered in a coated, protected form compared with traditional, uncoated ones. Here we discuss additional in vitro data of resistance of the same bacteria to gastric juice, human bile and pancreatic secretion and correlate them with the results of in vivo gut colonization. PMID- 21637031 TI - Effect of 25 weeks probiotic supplementation on immune function of HIV patients. AB - INTRODUCTION: Studies with a follow-up of < 8 weeks have indicated immune preserving effects of yogurt probiotic supplementation among HIV patients. To evaluate the impact of 25 weeks use of probiotics, a randomized, double blind, controlled study was undertaken on 65 women who were naive to anti-retroviral treatment. RESULTS: Ten participants were excluded post-randomization due to non eligibility. Thirty participants were assigned placebo, of whom 25 completed the study versus 19 of 25 completing the study in the probiotics group (p = 0.5). From baseline to 10 weeks follow-up, the CD4 count declined on average 3 CD4 cells/MUl (95% Confidence Interval: -97; 91) with placebo versus an increase of 50 cells/MUl (95% CI: -61; 162) with probiotics (p = 0.5). From baseline to 25 weeks, the CD4 count increased with 19 cells/MUl (95% CI: -90; 129) in the placebo group versus 46 cells/MUl (95% CI: -100; 192) with probiotics (p = 0.8). No differences in immune markers, diarrhea incidence or adverse events were observed. DISCUSSION: Lactobacillus GR-1 and RC-14 may be safely consumed at 2 x 10(9) CFU/day by moderately immune compromised HIV patients but this did not universally preserve immune-function. PATIENTS AND METHODS: Women were randomized to receive oral capsules containing Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 (2 x 10(9) colony forming units) or placebo twice daily for 25 weeks. The CD4 count and immune markers (IgG, IgE, IFNgamma and IL 10) were measured at baseline and during follow-up, the occurrence of diarrhea was reported daily. PMID- 21637032 TI - A more perfect union. PMID- 21637033 TI - Purpose of recently detected inhibitory domain of the Helicobacter pylori protein CagA. AB - Helicobacter pylori infection is the most common bacterial infection worldwide and is strongly associated with gastric oncogenesis. Recently, we discovered that the H. pylori protein CagA, a risk factor for carcinogenesis, consists of two distinct membrane-targeting domains. The C-terminal membrane-binding domain induces host cell responses associated with a high oncogenic potential. The N terminal membrane-targeting domain, however, localizes to a different membrane substructure at the site of newly formed cell-cell contacts thereby diminishing the effects of C-terminal signaling motifs on host cell physiology. This inhibitory function may allow H. pylori to establish a colonization niche in the host by maintaining the host epithelial architecture and thus decreasing the oncogenic potential as a side effect. From a bacterial standpoint, however, its main purpose maybe is to translocate the CagA protein via the type IV secretion apparatus into host epithelial cells. PMID- 21637034 TI - Responders and non-responders to probiotic interventions: how can we improve the odds? AB - As with many clinical studies, trials using probiotics have shown clearly that some patients benefit from the treatment while others do not. For example if treatment with probiotics leads to 36% cure rate of diarrhea, why did the other 64% not have the same result? The issue is important for human and indeed experimental animal studies for two main reasons: (i) Would changing the design of the study result in more subjects responding to treatment? (ii) If a subject does not respond what are the mechanistic reasons? In order to tackle the issue of responders and non-responders to therapy, a workshop was held by the International Scientific Association for Probiotics and Prebiotics (ISAPP). The outcome was four recommendations. 1. Clearly define the end goal: this could be supporting a health claim or having the highest clinical effect and impact. 2. Design the study to maximize the chance of a positive response by identifying precise parameters and defining the level of response that will be tested. 3. Base the selection of the intervention on scientific investigations: which strain(s) and/or product formulation should be used and why. 4. Carefully select the study cohort: use biological or genetic markers when available to stratify the patient population before enrollment and decide at what point intervention will provide the best outcome (for example, in acute phase of disease, or during remission, with or without use of pharmaceutical agents). By following these recommendations and selecting an appropriate primary outcome, it is hoped that clinical data will emerge in the future that expands our knowledge of which probiotics benefits which subjects and by what mechanism. PMID- 21637035 TI - Current level of consensus on probiotic science--report of an expert meeting- London, 23 November 2009. AB - The present paper summarizes the consensus views of a group of 9 European clinicians and scientists on the current state of scientific knowledge on probiotics, covering those areas where there is substantial evidence for beneficial effects and those where the evidence base is poor or inconsistent. There was general agreement that probiotic effects were species and often strain specific. The experts agreed that some probiotics were effective in reducing the incidence and duration of rotavirus diarrhoea in infants, antibiotic-associated diarrhoea in adults and, for certain probiotics, Clostridium difficile infections. Some probiotics are associated with symptomatic improvements in irritable bowel syndrome and alleviation of digestive discomfort. Probiotics can reduce the frequency and severity of necrotizing enterocolitis in premature infants and have been shown to regulate intestinal immunity. Several other clinical effects of probiotics, including their role in inflammatory bowel disease, atopic dermatitis, respiratory or genito-urinary infections or H.pylori adjuvant treatment were thought promising but inconsistent. PMID- 21637036 TI - Listeria monocytogenes: at the coalface of host-pathogen research. AB - Listeria monocytogenes is a highly adaptable food-borne pathogen that causes the life threatening illness listeriosis in infected individuals. Within the host this bacterium invades cells, escapes into the host cell cytosol and replicates intracellularly. To achieve this L. monocytogenes has evolved a sophisticated set of molecular weaponry that allows it to interact with and manipulate the cell biology of the host to its own advantage. Many of these interactions are well understood, putting this pathogen at the forefront of host-pathogen research, but fascinating new interactions are still emerging. The seventeenth International Symposium on Problems of Listeriosis (ISOPOL) was held in Portugal (Porto) in May of this year and this report describes some of the exciting developments that were presented at the meeting. The report focuses on developments in understanding the molecular interactions between L. monocytogenes and the host; it describes novel uses for L. monocytogenes as an anti-cancer treatment; and it describes some innovative uses of transcriptional profiling and reporter gene fusions that are helping illuminate our understanding of the basic biology of this important pathogen. PMID- 21637037 TI - Environmental epigenetic transgenerational inheritance and somatic epigenetic mitotic stability. AB - The majority of environmental factors can not modify DNA sequence, but can influence the epigenome. The mitotic stability of the epigenome and ability of environmental epigenetics to influence phenotypic variation and disease, suggests environmental epigenetics will have a critical role in disease etiology and biological areas such as evolutionary biology. The current review presents the molecular basis of how environment can promote stable epigenomes and modified phenotypes, and distinguishes the difference between epigenetic transgenerational inheritance through the germ line versus somatic cell mitotic stability. PMID- 21637039 TI - Current world literature. PMID- 21637041 TI - Bibliography. Speech therapy and rehabilitation. Current world literature. PMID- 21637042 TI - Benefits of hybrid SPECT/CT for (111)In-oxine- and Tc-99m-hexamethylpropylene amine oxime-labeled leukocyte imaging. AB - BACKGROUND: This retrospective study assessed the effect of the addition of SPECT/CT imaging on the diagnostic accuracy of labeled leukocyte scintigraphy. METHODS: Leukocyte scans of 134 patients performed between December 2005 and December 2009 were reviewed. All patients underwent whole-body planar and SPECT/CT imaging with either (111)In-oxine- or Tc-99m-hexamethylpropylene amine oxime (HMPAO)-labeled leukocytes. Leukocyte imaging identified a total of 221 lesions. Based on additional diagnostic tests and clinical follow-up, a final diagnosis was established in 115 patients. RESULTS: A total of 113 focal lesions with an established final diagnosis were detected on imaging in bone (n = 43), soft-tissues (n = 34), vascular grafts (n = 19), and other surgical implants (n = 17), whereas 26 scans resulted negative. Overall, leukocyte scanning including SPECT/CT yielded sensitivity, specificity, positive and negative predictive values of 87.5%, 85.3%, 83.6%, and 88.9%, respectively. As compared to planar imaging and SPECT, SPECT/CT imaging significantly increased the number of correctly identified lesion locations and improved overall reader confidence in 77 (68%) and 71 (63%) of 113 focal lesions, respectively (P < 0.001). Significant differences in scan accuracy were neither observed between In-oxine- or Tc-99m hexamethylpropylene amine oxime-labeled leukocyte studies, nor between scans obtained with or without antibiotic treatment. CONCLUSIONS: Hybrid SPECT/CT leukocyte imaging has incremental value over planar imaging with SPECT because of improved diagnostic accuracy of lesion identification and reader confidence. PMID- 21637043 TI - Tc-99m MAA findings in dilated cardiomyopathy with partial anomalous venous connections. AB - Tc-99m MAA showed asymmetric uptake in the lung field in a 21-year-old man with dilated cardiomyopathy. CT revealed partial anomalous venous connections in the left upper lobe. Angiogram of the left pulmonary upper lobe showed all the contrast material drained into the left vertical vein. The possible cause of relative increase in the left upper lobe blood flow is that right pulmonary blood flow is slowed by the high pressure in the left atrium due to dilated cardiomyopathy, whereas the flow from the left upper lobe drains into the superior vena cava which has less pressure than left atrium. PMID- 21637044 TI - Primary peripheral T-cell lymphoma of the colon mimics inflammatory bowel disease: a potential pitfall with F-18 FDG PET/CT imaging. AB - We present a case of a woman with ulcerative colitis who was referred for an F-18 FDG PET/CT after a suspicious colonoscopy and biopsy. PET/CT showed multiple elongated foci of intense FDG uptake in the colon, a pattern commonly reported with inflammatory bowel disease; however, the possibility of lymphoma was raised. Reevaluation of the biopsies with immunohistochemistry and molecular studies revealed a peripheral T-cell lymphoma and the patient was treated with a total colectomy. In this case, F-18 FDG PET/CT was very useful in determining the extent of the disease. This case also highlights a potential pitfall whereby the tubular and multifocal appearance of this lymphoma subtype can mimic the appearance of bowel involved by inflammatory bowel disease especially on novel techniques such as PET/CT enterography and PET/CT colonography. PMID- 21637045 TI - Methylene diphosphonate uptake in a case of vertebral hemangioma: demonstration with hybrid SPECT/CT. AB - Osseous hemangiomas are usually asymptomatic. Symptomatic hemangiomas are rare and represent approximately 1% of all cases. Hemangiomas usually present as photopenic defects in methylene diphosphonate (MDP) bone scintigraphy. MDP uptake in vertebral hemangiomas is extremely rare. We report a 20-year-old woman who presented with upper back pain. MDP bone scan showed focus of MDP uptake in the seventh dorsal vertebra. SPECT/CT localized the uptake to vertebral body and transverse process with CT findings suggestive of hemangioma. Further, MRI also confirmed hemangioma in the same location. PMID- 21637046 TI - Primary umbilical neoplasm demonstrated on FDG PET/CT. AB - Primary umbilical neoplasm is very rare, even though there are scattered case reports about the umbilical metastases known as Sister Mary Joseph's nodules. A 51-year-old man presented with an umbilical mass. Surgical biopsy showed infiltrating neoplasm with glandular and focal papillary architecture. FDG PET/CT demonstrated a focus of abnormal uptake located in the umbilicus, without evidence of a primary tumor elsewhere. Further investigations with EGD, colonoscopy, and cystoscopy were all negative for a neoplasm. The whole-body FDG PET/CT is an ideal imaging modality for evaluation of an umbilical lesion, especially identification or exclusion of a primary neoplasm elsewhere. PMID- 21637047 TI - Incidental finding of autosomal dominant polycystic kidney disease with liver involvement on Tc-99m sestamibi scintigraphy. AB - Autosomal dominant polycystic kidney disease is a systemic hereditary disease characterized by renal cysts and sometimes involvement of the liver. We present a 65-year-old woman with autosomal dominant polycystic kidney disease on regularly hemodialysis who recently experienced intermittent right upper quadrant abdominal pain and elevated intact parathyroid hormone for more than a year. She was referred for double-phase Tc-99m sestamibi scintigraphy, under the impression of hyperparathyroidism. Apart form increased uptake in the right thyroid bed, the images showed a large photon-deficient area in the upper portion of the abdomen corresponding to the liver. PMID- 21637048 TI - Unusual parathyroid adenoma avid for both Tc-99m sestamibi and I-123 iodine due to acquisition protocol. PMID- 21637049 TI - Synchronous second primary neoplasms detected by initial staging F-18 FDG PET/CT examination in patients with non-Hodgkin lymphoma. AB - PURPOSE OF THE REPORT: Cases of synchronous non-Hodgkin lymphoma (NHL) and second primary carcinoma in previously untreated immunocompetent patients are relatively rare. The aim of this part of our prospective study was to a revealed 2-F-18 fluoro-2-deoxy-D-glucose (F-18 FDG) positron emission tomography (PET)-positive lesion in an extranodal organ suggestive of second primary neoplasm in newly diagnosed NHL patients. MATERIALS AND METHODS: A total of 209 patients with NHL underwent initial staging F-18 FDG PET/computed tomography (CT). The finding was assessed by a radiologist, nuclear medicine physician, and hematologist. In 6 suspicious cases (2.9%) of second neoplasm, the decision was made to perform further investigations before lymphoma therapy. RESULTS: Two patients were diagnosed with colorectal carcinoma, 1 with esophageal adenocarcinoma, 1 with invasive ductal breast carcinoma, 1 with medullary thyroid carcinoma, and 1 with squamous cell lung carcinoma. In 5 of the 6 patients, the second solid tumor was completely asymptomatic and revealed only by F-18 FDG PET/CT examination. CONCLUSIONS: We conclude that in patients with NHL, appropriate imaging, clinical, and histologic analysis of organ lesions detected by F-18 FDG PET/CT will occasionally demonstrate significant synchronous neoplasms. PMID- 21637050 TI - FDG PET/CT in detection of adrenal metastasis in patients with renal cell carcinoma. AB - AIM: Adrenal metastasis in renal cell carcinoma (RCC) is uncommon. The present study was aimed to evaluate the role of fluoro-deoxy-glucose positron emission tomography (FDG PET)/computed tomography (CT) in the detection of adrenal metastasis in patients with RCC. MATERIALS AND METHODS: A total of 9 FDG PET/CT scans were performed in 4 patients (3 males, 1 female; median age: 53.5 years, range: 52-73 years). Of 9 PET/CT scans, 2 scans were done for initial staging, 2 for restaging, and 5 for evaluation of treatment response. PET/CT image interpretation and analysis was performed qualitatively (visually) and semiquantitatively. Absence of uptake in postchemotherapy PET/CT scan was considered as complete response and a fall in SUV by more than 50% of baseline value, as significant response. RESULTS: Patients 1 and 2 underwent staging FDG PET/CT. Patient 1 had undergone left radical nephrectomy 13 years back for left sided RCC. Current FDG PET/CT revealed appearance of right RCC with adrenal metastasis. PET/CT of patient 2 demonstrated right adrenal and distant metastases. PET/CT done for restaging in patient 3, revealed right adrenal and abdominal lymph nodal metastasis. In patient 4, PET/CT revealed solitary adrenal metastasis. Magnetic resonance imaging findings were consistent with adrenal metastasis in this patient. Patient 1 received sunitinib and interferon chemotherapy along with radiotherapy and showed partial response on PET/CT scans done at 6, 17, 23, and 33 months from the initial study. PET/CT of patient 3 demonstrated the progression of disease (nonresponder). CONCLUSION: FDG PET/CT appears to be useful for staging, treatment response evaluation, and recurrence detection in patients with RCC having adrenal metastasis. PMID- 21637051 TI - A comparative study of fused FDG PET/MRI, PET/CT, MRI, and CT imaging for assessing surrounding tissue invasion of advanced buccal squamous cell carcinoma. AB - PURPOSE: This study aimed to evaluate the diagnostic value of fused fluorodeoxyglucose positron emission tomography and magnetic resonance imaging (PET/MRI) compared with PET/computed tomography (CT), MRI, and CT in assessing surrounding tissue invasion of advanced buccal squamous cell carcinoma (BSCC). MATERIALS AND METHODS: PET/CT and MRI were performed in 17 consecutive patients with suspected masticator space invasion of BSCC from CT images. Attenuation corrected PET and head and neck MRI datasets were registered. For pathologic correlation, 4 regions of interest were examined, including the maxilla, mandible, pterygoid, and masseter muscle. The tumor maximal diameter, measured by different imaging modalities, was correlated with pathology results. RESULTS: All PET/MRI fusions were verified as well matched using specific anatomic criteria. For pathology results, 1 patient had inflammation only, 1 had spindle cell cancer, and 15 had squamous cell cancer. Of 64 regions of interest, 20 (31.3%) harbored tumor invasion. The likelihood ratio was highest in fused PET/MRI (42.56) compared with PET/CT (25.02), MRI (22.94), and CT (8.6; all P < 0.05). The sensitivity and specificity of fused PET/MRI were also highest among the 4 modalities (90.0%/90.9%, 80.0%/84.1%, 80.0%/79.5%, and 55.0%/81.8%, respectively). The level of confidence was higher in fused PET/MRI or MRI than in PET/CT or CT (85.9%, 85.9%, 70.3%, 73.4%, respectively). The maximal lesion size was 3.0 to 6.0 cm in the pathology specimen. Regression analysis showed better agreement between fused PET/MRI and pathology results. CONCLUSIONS: Fused PET/MRI is more reliable for focal invasion assessment and tumor size delineation in advanced BSCC compared with PET/CT, MRI, and CT. PET/CT has the lowest confidence level, which may limit its use in the clinical setting. PMID- 21637052 TI - PET/CT allows stratification of responders to neoadjuvant chemotherapy for high grade sarcoma: a prospective study. AB - PURPOSE: The aim of the present study was to determine whether metabolic reduction is capable of reflecting the histopathologic response and outcome after neoadjuvant chemotherapy in patients with high-grade sarcoma. PATIENTS AND METHODS: Forty-two patients with histologically proven high-grade sarcoma underwent neoadjuvant chemotherapy followed by surgical resection. Quantitative F 18 fluorodeoxyglucose (F-18-FDG) positron emission tomography (PET)/computed tomography scans were acquired before and after the first cycle and after completion of neoadjuvant chemotherapy. Standardized uptake values (SUVs) and metabolic reduction rates were compared with histopathologic response, progression-free survival, and overall survival. RESULTS: Baseline SUVmax was 10.9 +/- 3.6 (range, 3.8-19.6). Therapeutic effect resulted in 10 patients (24%) with a satisfactory response and in 32 patients (76%) with an unsatisfactory response after completion of neoadjuvant chemotherapy. The SUV decreased to 7.8 +/- 3.4 after the first cycle (t1) of chemotherapy and to 5.2 +/- 3.4 after completion (t2) of chemotherapy. Histopathologic response and percentage SUV (t2) reduction rate were independent predictors of progression-free survival and overall survival in the multivariate analyses. CONCLUSION: Metabolic reduction after neoadjuvant chemotherapy evaluated by F-18 FDG PET or computed tomography can be used for stratification of the histopathologic response in patients with high-grade sarcoma. PMID- 21637053 TI - Clinical role of FDG PET/CT for methotrexate-related malignant lymphoma. AB - Methotrexate-related malignant lymphoma (MTX-RML) is a type of therapy-related lymphoma, and it often occurs in patients with rheumatoid arthritis. The most distinctive characteristic of MTX-RML is a quick response to withdrawal of MTX. However, because there is a risk of recurrence without a distinctive indicator of disease, close follow-up is needed. We present F-18 2-fluoro-2-deoxyglucose (FDG) postitron emission tomography (PET) or computed tomography (CT) images of MTX-RML along with the characteristic clinical presentation of MTX-RML. FDG PET/CT has the advantage of being able to detect malignant lymphoma in patients who have undergone MTX treatment. After withdrawal of MTX, FDG uptake decreases along with a reduction in the volume of lesions. Although recurrent lesion develops independent to the initial FDG PET/CT findings, FDG PET/CT is useful for early detection of unexpected recurrent lesions. FDG PET/CT allows for the assessment of malignant lymphoma and recurrent lesions in patients who received MTX therapy, which is crucial for the management of MTX-RML. PMID- 21637054 TI - The value of fused SPECT/CT in the evaluation of solitary skull lesion. AB - AIM: The aim of this study was to evaluate the role of single photon emission computed tomography and/or computed tomography (SPECT/CT) in differentiating metastatic from benign solitary skull lesions. MATERIALS AND METHODS: Consecutive patients who had a SPECT/CT of the head subsequent to a whole-body bone scan (WBS) for the evaluation of a single skull lesion were selected. A single skull lesion on the WBS was further evaluated with SPECT/CT to characterize the lesion. The results of the SPECT/CT were correlated with other radiologic examinations performed within 2 weeks. An average follow-up interval after the SPECT/CT was 8.9 months to correlate with additional radiologic imaging studies and clinical information. RESULTS: A total of 19 lesions in 19 patients were seen on the WBS and 2 additional lesions on the SPECT/CT. All lesions demonstrated focal increased tracer uptake. The SPECT/CT correctly identified 3 out of 3 metastatic lesions and 12 out of 17 benign lesions, that is 71% of lesions were correctly classifised as metastatic or benign lesions. Only 1 patient was classified incorrectly as metastatic lesion with SPECT/CT when it was proven benign by other imaging modalities and follow-up. The sensitivity, specificity, positive and negative predictive values of SPECT/CT images in identifying metastatic lesions were 100%, 92%, 75%, and 100%, respectively. Five lesions remained indeterminate even after the SPECT/CT interpretation and were confirmed benign by other imaging modalities. CONCLUSION: SPECT/CT can help identify benign versus metastatic solitary skull lesions in most of the patients with high sensitivity and specificity. PMID- 21637055 TI - Whole-body gallium-67 scintigraphic findings in IgG4-related disease. AB - PURPOSE: To clarify the features of gallium-67 (Ga-67) imaging typical of IgG4 related disease. METHODS: We retrospectively investigated 14 patients diagnosed with IgG4-related disease who underwent Ga-67 scintigraphy in our hospital between January 2005 and May 2010. Of these, 13 patients who underwent gallium scintigraphy before steroid therapy were enrolled in this study. The patient population comprised 11 men and 2 women with age ranging from 47 to 76 years (mean age, 61.9 years). RESULTS: Among the 13 patients, significant accumulation of Ga-67 was detected in the bilateral pulmonary hila in 10 patients (77%), pancreas in 10 (77%), salivary glands in 7 (54%), lacrimal glands in 7 (54%), periaortic lesions in 2 (15%), and lung parenchyma in 1 (8%) patient. High accumulation of Ga-67 in the salivary glands was observed in the parotid glands in 3 cases and in the submandibular glands in 6 cases, with the following pattern: normal parotid gland uptake and symmetrical submandibular gland uptake in 2 cases; symmetrical high accumulation in both parotid and submandibular glands in 1 case; symmetrical high accumulation in parotid glands and normal submandibular gland uptake in 1 case; symmetrical uptake by parotid glands and asymmetric uptake by submandibular glands in 1 case; normal parotid gland uptake and asymmetric submandibular gland uptake in 1 case; and asymmetric parotid gland uptake and symmetrical parotid gland uptake in 1 case. CONCLUSION: Characteristic patterns of gallium uptake would be helpful for diagnosis, detection of involved lesions, and differential diagnosis in patients with IgG4-related disease to avoid unnecessary surgery. PMID- 21637056 TI - Risk evaluation for coronary artery disease in patients with impaired glucose tolerance after a successful coronary intervention. AB - PURPOSE: Patients with coronary artery disease (CAD) often have risk factors that may influence endothelial function. The purpose of this study was to evaluate the endothelial function and its association with coronary risk factors after percutaneous coronary intervention (PCI). MATERIALS AND METHODS: A total of 14 patients with impaired glucose tolerance and CAD underwent positron emission tomography with N-13 ammonia to measure myocardial blood flow (MBF) at rest and during a cold pressor test (CPT), to estimate endothelial function as a percent increase (%increase) of MBF. The results were compared among normal segments (normal), reperfused segments with PCI (PCI), and nonculprit CAD segments without PCI (non-PCI). Correlations between the %increase and major risk factors were also investigated. RESULTS: CPT induced significant increase in MBF in all groups. The %increase of normal, non-PCI, and PCI groups were 33% +/- 22%, 21% +/ 23%, and 26% +/- 23%, respectively. Comparison with risk factors demonstrated significant correlations only in the non-PCI group. Specifically, there were negative correlations between %increase and fasting blood sugar (r = -0.64, P < 0.05), hemoglobin A1c (r = -0.74, P < 0.05), total cholesterol (r = -0.87, P < 0.05), triglyceride (r = -0.71, P < 0.05), and low-density lipoprotein cholesterol (r = -0.92, P < 0.005), respectively. CONCLUSIONS: Although impaired glucose tolerance patients with a PCI-treated coronary stenosis showed preserved response to CPT, the %increase negatively correlated with risk factors in the non PCI segments. Therefore, coronary risk factors may affect CAD lesions in PCI treated patients. PMID- 21637057 TI - F-18 FDG PET for evaluation of bone marrow involvement in non-Hodgkin lymphoma: a meta-analysis. AB - BACKGROUND: In recent years, the use of F-18 fluorodeoxyglucose (FDG) positron emission tomography/computed tomography (PET/CT) has become widespread for the staging of lymphoma. In non-Hodgkin lymphoma (NHL), the bone marrow (BM) involvement is a sign of extensive disease, and the iliac crest BM biopsy (BMB) is the established method for the detection of BM infiltration. However, iliac crest BMB is associated with a high rate of false-negative results. We assess the ability of FDG PET or PET/CT scan to ascertain the presence of BM involvement in aggressive and indolent NHL. METHODS: The authors conducted a systematic MEDLINE search of articles published (last update, May 2010). Two reviewers independently assessed the methodological quality of each study. A meta-analysis of the reported sensitivity and specificity of each study was performed. RESULTS: Eight studies met the inclusion criteria. The studies had several design deficiencies. Pooled sensitivity and specificity for the detection of non-Hodgkin aggressive lymphoma were 0.74 (95% CI, 0.65-0.83) and 0.84 (95% CI, 0.80-0.89), respectively. Pooled sensitivity and specificity for the detection of non-Hodgkin indolent lymphoma were 0.46 (95% CI, 0.33-0.59) and 0.93 (95% CI, 0.88-0.98), respectively. CONCLUSIONS: The diagnostic accuracy of FDG PET or PET/CT scans was slightly higher but without significant statistical difference (P = 0.1507) in patients with non-Hodgkin aggressive lymphoma as compared with those with non Hodgkin indolent lymphoma. The sensitivity to detect indolent lymphoma BM infiltration was low for FDG PET or PET/CT. PMID- 21637058 TI - Atypical femoral head osteonecrosis: a case report. AB - We report the case of a 65-year-old man presented with polymyalgia rheumatica. After a week on corticosteroid (40 mg/d), pain was relieved rapidly. Bone scan, requested to precisely localize osteoarticular lesions, showed high uptake in the external aspect of the head of left femur. In the clinical setting, bone scan and MRI appearances are suggestive of osteonecrosis, probably of recent onset. The final diagnosis of atypical necrosis of the femoral head, most probably secondary to corticoid therapy, was thus established. PMID- 21637059 TI - A unique case of thyroid dysgenesis in a neonate. AB - Neonates with congenital hypothyroidism often demonstrate ectopic thyroid tissue. However, 2 sites of abnormally located thyroid tissue are exceptionally rare with only one other published case on a neonate. The authors present a unique case of a neonate who was found to have congenital hypothyroidism on laboratory screening; Tc-99m pertechnetate scintigraphy demonstrated 2 sites of abnormal uptake: one at the base of the tongue and the other in the midline at the expected location of the thyroid bed. PMID- 21637060 TI - F-18 FDG PET/CT as a crucial guide toward optimal treatment planning in a case of postirradiation sarcoma 10 years after primary bone lymphoma of the pelvis. AB - Ten years after chemoradiation for primary lymphoma of the left pelvic bone, a 38 year-old man presented with a 4-month history of gradually increasing pain in his left upper leg and thigh. Initial radiographs and contrast-enhanced magnetic resonance imaging were consistent with recurrent lymphoma, infection, or postirradiation sarcoma. Subsequent F-18 fluorodeoxyglucose positron emission tomography/computed tomography demonstrated a focal area of F-18 fluorodeoxyglucose-avidity within the previously irradiated bone consistent with tumor and the location was confirmed by advanced magnetic resonance imaging techniques and histopathology, thus optimizing treatment planning. PMID- 21637061 TI - An unusual unilocular mesothelial cyst of the pleura: a case report. AB - A 79-year-old woman was found to have an abnormal shadow on chest radiography. Computed tomography demonstrated a pleural mass. The F-18 fluorodeoxyglucose positron emission tomography (FDG PET) was performed to determine whether the pleural mass was benign or malignant. The histologic examination of the resected mass showed a unilocular mesothelial cyst of the pleura. The FDG PET findings of a mesothelial cyst of the pleura have not yet been previously reported. The FDG PET findings of a mesothelial cyst in the pleura reflected the microscopic findings of the resected mass. The FDG PET findings, therefore, seem to be useful in the diagnosis of mesothelial cysts. PMID- 21637062 TI - Synchronous chronic middle cerebral artery occlusion and ipsilateral dural arteriovenous fistula. AB - Dural arteriovenous fistulas (dAVFs) are a complex family of vascular malformations. Most series discuss features in isolation of other vascular pathologies, despite evidence that dAVFs may arise in a setting of elevated angiogenic growth factors related to regional hypoxia. We describe a 72-year-old man with ischemic symptoms secondary to middle cerebral artery stenosis and an incidental ipsilateral Borden I dAVF. Balloon-occlusion single photon emission computed tomography demonstrated reversible ischemia within the basal ganglia consistent with fistula-related "steal" phenomenon. This case supports the hypothesis that dAVFs may arise secondary to angiogenic growth factors related to regional tissue hypoxia. PMID- 21637063 TI - A rare case of Tolosa-Hunt syndrome imaged with FDG PET/CT and MRI. AB - We report on the imaging findings of Tolosa-Hunt syndrome in a 59-year-old patient. Clinical findings included periorbital pain, ptosis, disordered eye movements, and blurred vision. Treatment with intravenous administration of steroid resolved all symptoms. Currently, magnetic resonance imaging plays a key role in the diagnosis of Tolosa-Hunter syndrome for locating the inflammatory tissue and follow-up. This case of Tolosa-Hunter syndrome with representative (FDG PET/CT) images may imply that FDG PET/CT is a useful tool in detecting and monitoring of this disease. PMID- 21637064 TI - Rapidly growing complex fibroadenoma with surrounding ductal hyperplasia mimics breast malignancy on serial F-18 FDG PET/CT imaging. AB - A 30-year-old woman was referred for an F-18 fluorodeoxyglucose (FDG) PET/CT to rule out lymphoma, and was found to have an incidental FDG-avid right breast nodule that grew significantly in size and FDG uptake on a subsequent scan, raising suspicion of a growing breast malignancy. Histologic evaluation showed a complex fibroadenoma with adenosis and surrounding ductal hyperplasia. Although variable F-18 FDG uptake in fibroadenomas has been described, a distinction between simple and complex fibroadenomas has not been made in the PET literature, even though complex fibroadenomas have a higher propensity to develop into malignancies. This case shows that a rapidly growing complex fibroadenoma can mimic a breast malignancy on serial F-18 FDG PET/CT scans, showing significant increase in both size and FDG-avidity on follow-up studies. PMID- 21637065 TI - Cardiac hydatid cyst mimicking malignancy. AB - A 50-year-old man presented with chest pain symptoms for 7 years. He did not report any other complaints. The diagnostic tests including physical examination, routine laboratory tests, chest x-ray, and electrocardiography were reported as normal. Cardiac MRI, however, revealed a 64 * 66 mm mass lesion in the left myocardium. He was referred to our unit for further tests. A PET/CT was carried out which demonstrated a minimally increased fluorodeoxyglucose uptake at the peripheral margins of the mass, which deemed to be suspicious for malignancy. The patient underwent a thoracotomy and the cardiac mass was excised. Histopathologic examination reported this lesion as a hydatid cyst. PMID- 21637066 TI - FDG PET/CT in a patient with spontaneous remission of methotrexate-associated lymphoproliferative disorders after interruption of methotrexate. AB - We report a case that fluorodeoxyglucose positron emission tomography (FDG PET)/computed tomography (CT) depicted systemically multiple lesions of methotrexate (MTX)-associated lymphoproliferative disorders. A 70-year-old man receiving MTX for rheumatoid arthritis complained of neck swelling. FDG PET/CT revealed multiple FDG-avid lesions in lymph nodes, lungs, bones, and muscles. Lesions in bones and muscles were not detected on X-CT. Final diagnosis of cervical node was confirmed as MTX-associated lymphoproliferative disorders. FDG PET/CT performed 148 days after discontinuing MTX demonstrated complete remission. PMID- 21637067 TI - Cutaneous sarcoidosis evaluated by FDG PET. AB - A 50-year-old man presented with initial complaints of diffuse skin pain and pruritus. Physical examination revealed scattered skin plaques and subcutaneous nodules with mild tenderness throughout the body. Skin biopsy demonstrated noncaseating epithelioid granulomas. Patient soon developed cough, fever with hot flashes, and shortness of breath on exertion. FDG PET/CT demonstrated diffuse cutaneous involvement throughout the body. Follow-up FDG PET/CT after treatment revealed a decrease in FDG uptake suggesting a good response to therapy. PMID- 21637068 TI - Usefulness of F-18 FDG PET/CT in the evaluation of neck swelling in polymorphic posttransplant lymphoproliferative disorder after renal transplantation. AB - A 38-year-old man with a history of renal transplantation presented with a 3 month history of bilateral neck swelling. Ultrasonography of the neck showed multiple enlarged lymph nodes at the bilateral neck. Histologic examination of a specimen from an excisional biopsy at right level II cervical lymph node revealed polymorphic posttransplant lymphoproliferative disorder. F-18 fluorodeoxyglucose (FDG) positron emission tomography/computed tomography scan was performed for further evaluation. The scan revealed marked enlargement and intense hypermetabolism of the lymph nodes along the bilateral cervical lymph node chains (Fig. 1). Several reports have described that diverse infectious or inflammatory disorders and malignant diseases show various degree of FDG uptake of cervical lymph nodes. This case illustrates that the possibility of posttransplant lymphoproliferative disorder should be considered in renal transplant patients with increased FDG uptake of cervical lymph nodes on F-18 FDG positron emission tomography/computed tomography scan. PMID- 21637069 TI - Malignant epithelioid hemangioendothelioma of the lower extremity: staging with F 18 FDG PET/CT. AB - A 36-year-old woman presented with a 5-month history of a growing left thigh mass, causing difficulty walking. Biopsy revealed a malignant epithelioid hemangioendothelioma (EHE), and the patient was referred for a staging F-18 FDG PET/CT that showed intense FDG uptake in the thigh mass, but no FDG-avid local lymph nodes or distant metastases. A few reports have noted the usefulness of F 18 FDG PET/CT imaging in the staging of EHEs in the lung, liver, and bone marrow. This rare study highlights the usefulness of F-18 FDG PET/CT in the staging of malignant EHE of the soft tissues of the extremities. PMID- 21637070 TI - FDG PET imaging in a patient with primary seminoma of the prostate. AB - Primary seminoma of the prostate is extremely rare. We describe a case of a 35 year-old man who presented with difficulty in urinating. Physical findings were unremarkable, but a rectal examination revealed a slightly hardened prostate with a nodule in the right lobe. Blood parameters were normal; however, blood chemistry revealed slightly elevated lactic dehydrogenase, and microscopic hematuria was evident. Serum prostate acid phosphate, carcinoembryonic antigen, and alpha-fetoprotein values were within normal limits. Both computed tomography and magnetic resonance images revealed a large mass in the prostate gland. A prostatic sarcoma or malignant lymphoma was initially suspected. A systemic survey using 18-fluoro-2-deoxyglucose (F-18 FDG) positron emission tomography revealed focal FDG uptake in the prostate gland. A prostatic biopsy of the mass was histologically diagnosed to reveal an extragonadal seminoma. PMID- 21637071 TI - Umbilical hernia simulating recurrent carcinoid on octreoscan. AB - Physiologic bowel activity on In-111 octreotide imaging is commonly seen. However, on occasion, normal bowel activity may mimic lesions in the abdomen, which can be very difficult to differentiate, particularly after surgical intervention. We report a case of focal bowel activity simulating a lesion in a patient who had an In-111 octreotide scan (Octreoscan), postoperatively, looking for recurrent carcinoid. SPECT/CT demonstrated that the uptake was localized in the anterior abdomen, and corresponded to a loop of bowel within a ventral hernia at the site of surgical incision. The correlation with CT as well as the precise localization made possible by the fusion images helped avoid misinterpretation of this finding as possible recurrent carcinoid. PMID- 21637072 TI - Hemosuccus pancreaticus in Glanzmann thrombasthenia: implication of scintigraphy in gastrointestinal bleeding (GIB) of unknown origin. AB - A 33-year-old man with Glanzmann thrombasthenia, enduring retractable melena, and with an active upper gastrointestinal bleeding (GIB) of unknown origin lasting intermittently for 6 months, was admitted. Abdominal ultrasound and computed tomography were not diagnostic. The patient underwent upper gastrointestinal endoscopy twice, both of which showed blood issuing forth from ampulla of vater and possible diagnoses of bleeding from papilla/hematobilia or hemosuccus pancreaticus were suspected. Digital subtraction angiography of celiac/superior mesenteric arteries was unremarkable. In GIB scintigraphy, a focus of activity appeared in the epigastric area early in the study (arrow), intensity of which increased gradually. Exploratory laparatomy confirmed the diagnosis of hemosuccus pancreaticus. Although the patient was doing well for few weeks after the surgery, he died 3 months later after an acute episode of severe GIB. PMID- 21637073 TI - Malignant extrarenal rhabdoid tumor of the spine: staging and evaluation of response to therapy with F-18 FDG PET/CT. AB - Malignant extrarenal rhabdoid tumor (ERRT) is a very rare type of soft-tissue sarcoma with a reported incidence of 0.3% of all soft-tissue sarcomas. Only 7 cases of spinal malignant ERRT have been reported in the literature, and to our knowledge, F-18 FDG PET/CT imaging for staging and evaluation of response to therapy for these tumors has not been previously described. This is a case of an 8-month-old boy with malignant ERRT of the spine, who was staged with F-18 FDG PET/CT, and had his tumor burden assessed with PET/CT after chemotherapy, which altered the subsequent chemotherapy regimen. PMID- 21637074 TI - Assessment of radiofrequency ablation of lung metastasis from colorectal cancer using dual time-point PET/CT. AB - A 70-year-old man with operated colorectal cancer relapsed with a solitary lung metastasis. Dual time-point PET/CT performed at 60' (standard images) and after 2 hours (delayed images) showed focal uptake in the lung nodule. A second uptake in the liver dome was also visualized only in the delayed images. Radiofrequency of the lung metastasis was performed by CT-fluoroscopy guide. A PET/CT 1 month after radiofrequency showed significant reduction of activity in the delayed images, and the uptake at 3 months was virtually normal. A fine-needle biopsy of the liver lesion confirmed the final diagnosis of metastasis from adenocarcinoma. PMID- 21637075 TI - A case of secondary hemochromatosis with high uptake of liver in F-18 FDG PET/CT imaging. AB - Hemochromatosis is an iron storage disorder with progressive and massive deposition of iron in the parenchymal cells of various organs. A 72-year-old woman with rheumatoid arthritis diagnosed with secondary hemochromatosis underwent F-18 fluorodeoxyglucose (FDG) positron emission tomography(PET)/computed tomography (CT) to search for malignancy. F-18 FDG PET/CT incidentally showed homogeneous and high FDG uptake (mean SUV, 4.8) in the liver with high density. FDG uptake seemed to be reflected by the functional abnormality of liver. Hemochromatosis should be considered when making a diagnosis of homogeneous high F-18 FDG uptake in liver with high density on CT image. PMID- 21637076 TI - Extraosseus uptake of F-18 fluoride in the primary malignancy and cerebral metastasis in a case of non-small-cell lung cancer. AB - A 68-year-old man with history of heavy smoking was admitted for increasing falls during the past 4 weeks. Chest x-ray revealed a right upper lobe mass. Biopsy demonstrated poorly differentiated non-small-cell carcinoma. F-18 fluoride positron emission tomography/computer tomography (PET/CT) was performed to evaluate bone metastasis. Review of the sectional PET images demonstrated extraosseous fluoride uptake in the primary lung mass, as well as ring-shaped fluoride uptake in the cerebral metastatic lesion. Neither of these lesions demonstrated calcifications on CT images. The patient received radiation treatment of the brain metastasis after F-18 fluoride PET/CT study. PMID- 21637077 TI - A child with Burkitt lymphoma with pleural, peritoneal, mesenteric, omental, and renal involvement: diagnostics by FDG PET/CT. AB - A 7-year-old boy presented with increasing abdominal girth and abdominal pain. Abdominal ultrasound revealed a pleural effusion and ascites. Computed tomography (CT) demonstrated a mass involving the peritoneum, mesentery, and omentum with marked ascites, mesenteric nodules, and renal involvement. Positron emission tomography (PET)/CT scan identified additional tumor within the pleura. PET/CT imaging was helpful in identifying the appropriate pleural mass biopsy site in this patient with Burkitt lymphoma. PMID- 21637078 TI - Syphilitic bone destruction on FDG PET/CT. AB - A 35-year-old man underwent PET/CT scan to evaluate suspected osseous metastases from a presumed tumor of unknown origin. PET images demonstrated multiple foci of increased FDG uptake in all extremities, corresponding to the osseous destruction on the concurrent CT images. However, no primary tumor was identified. Instead, the bone lesions were subsequently proven due to syphilis infection. PMID- 21637079 TI - Current readings in nuclear medicine. PMID- 21637081 TI - Editorial comment: spondyloarthritides. PMID- 21637082 TI - Dual-energy X-ray absorptiometry in clinical practice: new guidelines and concerns. AB - PURPOSE OF REVIEW: To summarize recent policy and guideline updates that have significant consequences for the clinical use of dual-energy X-ray absorptiometry (DXA) in the diagnosis and treatment of osteoporosis and, thus, the prevention of fractures and associated morbidity and mortality. RECENT FINDINGS: Recent policy trends have sought to reverse reimbursement declines for DXA services, whereas updated guidelines have attempted to restrict the candidates for bone density testing. Recent literature reflects the ongoing controversy of repeat DXA testing. SUMMARY: Patient access to DXA scans has been threatened by declining reimbursement and, therefore, access to diagnosis and fracture prevention. There have been successful efforts to reverse this trend, but the future remains uncertain. The complexities and omissions of updated guidelines for obtaining DXA testing may serve again to restrict initial access, and the recent controversy of repeat DXA testing may make monitoring results of therapy more difficult. PMID- 21637083 TI - Bibliography. Spondyloarthropathies. Current world literature. PMID- 21637084 TI - Transformation and the power of intention. PMID- 21637085 TI - The reliability and validity of the Turkish version of the constipation risk assessment scale. AB - The aim of this study is to translate into the Turkish language, and test the reliability and validity, of the Turkish version of the Constipation Risk Assessment Scale (CRAS). This study consisted of 245 adult in-patients who were hospitalized in the medical and surgical clinics of Celal Bayar University Hospital in January through May 2007. The patients were categorized into two groups (constipated and not constipated) according to Rome II criteria. All participants were assessed with the CRAS. The CRAS was retested on 32 patients selected randomly from among the initial constipated group (n =152). The statistical analysis consisted of reliability and validity analyses. Test-retest comparison and internal consistency were used to assess the reliability of the instrument. Divergence and known groups approaches were used to test for construct validity. Correlation analysis using the Pearson's coefficient was conducted to assess the test-retest. For testing of the criteria and known groups, Student's t test and Mann-Whitney U test were used. Cronbach's = value for the constipated respondents was r = 61.9. According to the effect size comparisons, the most effective variable on the CRAS score was perception of constipation risk requirement. The overall score and subsection score correlations were also found acceptable (r = 0.47-0.57). PMID- 21637086 TI - Comparisons of screening colonoscopy performed by a nurse practitioner and gastroenterologists: a single-center randomized controlled trial. AB - Several barriers to colorectal cancer screening have been identified including limited access to trained endoscopists and highlight insufficient capacity to meet projected demand for colonoscopies. Two European studies have found that nonphysician providers can perform colonoscopies as safely and accurately as physicians. Training nurse practitioners (NP) to perform colonoscopy may be an effective strategy to increase access. The goal of this study was to compare accuracy, safety, and patient satisfaction in screening colonoscopy performed by board certified gastroenterologists (GI-MD) and a gastroenterology trained nurse practitioner (GI-NP). A consecutive sample of average risk participants referred for screening colonoscopy was randomized to have their procedure performed by either a GI-MD (n = 100) or a GI-NP (n = 50). Participants completed a preprocedure and postprocedure questionnaire. Endoscopists completed a postprocedure questionnaire. Cecal intubation rates, duration of procedure, sedative, and analgesic use, and patient reported procedural pain scores were equivalent among the groups. The GI-NP group had a higher adenoma detection rate compared with the combined GI-MD groups (42% and 17%, respectively, p = .0001) and a higher satisfaction score when compared with the combined GI-MD groups (mean 5.9 +/- 13.81 and 8.6 +/- 16.11, respectively, p = .042; visual analog scale 0-100 mm, "0" = completely satisfied, "100" = completely dissatisfied). There were no immediate complications reported in any group. The properly trained GI-NP in our study performed screening colonoscopy as safely, accurately, and satisfactorily as the GI-MDs. Using well-trained NPs for screening colonoscopy can be an effective strategy to increase access to colorectal screening. PMID- 21637087 TI - Validating patient safety in the endoscopy unit using the joint commission standards. AB - Patient safety has gained increasing interest and importance in the endoscopy suite. To maintain standards in patient identification, confidentiality, medication safety, and specimen handling while still meeting The Joint Commission or other agency requirements, all employees must be part of this process. This article demonstrates ways to accomplish this goal, using audits of safety standards that increase staff involvement. A sample audit tool is provided, along with methods to help all employees feel empowered to meet standards and report any patient safety issues. PMID- 21637088 TI - Celiac disease: implications for patient management. AB - Celiac disease is an autoimmune disorder that is known specifically for causing inflammation of the mucosa in the small intestine. Through multiple diagnostic and screening tools such as small intestinal biopsy sample, serological testing, and human leukocyte antigen testing, healthcare providers can diagnose this disease that contains components related to genetic predisposition and intake of gluten proteins found in wheat, barley, and rye. There are some who believe that having an autoimmune disease may predispose one to acquiring another disease. With patients experiencing mostly diarrhea, abdominal pain, and weight loss, the implementation of a gluten-free diet is the treatment that healthcare providers recommend. Through monitoring gluten intake and providing nutritional supplementation, those diagnosed with celiac disease can lead a relatively normal life without complications. With celiac disease affecting all age ranges in the population, and with a documented higher frequency, there is a growing awareness in society that can be easily seen in grocery stores, restaurants, and food manufacturers. PMID- 21637090 TI - Medical mission volunteering: Ecuador. AB - This article discusses a volunteer medical mission of 50 people traveling to Ecuador. Included were surgeons, an anesthesiologist, a gastroenterologist, and an ear, nose, and throat surgeon as well as certified registered nurse anesthetists, registered nurses for pre-, intra-, and postrecovery, a certified registered gastroenterology nurse, and helpers. The focus is on the people involved, starting as strangers, working together, crossing bridges, and helping others in a third world country. PMID- 21637091 TI - Perceived self-efficacy among liver transplant candidates and recipients. AB - Self-efficacy is defined as one's belief that one can perform the necessary behaviors to achieve one's goals. This belief is one of the most powerful motivational predictors of how well a person will perform at almost any endeavor. Limited information is available, however, about self-efficacy in the organ transplantation population. This study aimed to identify the self-efficacy belief among liver transplant candidates and recipients and compare these results with demographic characteristics. The authors analyzed 100 patients who were candidates and recipients for liver transplantation in a Brazilian center. The average self-efficacy score for the candidates was significantly lower than for the recipients. Future investigation is necessary to explore possible interventions to be taken before and after liver transplantation that could influence self-efficacy of the organ transplantation patients. PMID- 21637092 TI - Flexible bronchoscopy with transbronchial needle aspiration confirmed by endobronchial ultrasound. PMID- 21637094 TI - Lusedra (fospropofol disodium). PMID- 21637095 TI - New insights into the haemodynamic effects of dipyridamole. PMID- 21637096 TI - Detection of left main coronary artery anomalous origin with low-dose multidetector computed tomography using prospective ECG gating. PMID- 21637099 TI - Simulator training to automaticity leads to improved skill transfer compared with traditional proficiency-based training: a randomized controlled trial. AB - OBJECTIVE: We hypothesized that novices will perform better in the operating room after simulator training to automaticity compared with traditional proficiency based training (current standard training paradigm). BACKGROUND: Simulator acquired skill translates to the operating room, but the skill transfer is incomplete. Secondary task metrics reflect the ability of trainees to multitask (automaticity) and may improve performance assessment on simulators and skill transfer by indicating when learning is complete. METHODS: Novices (N = 30) were enrolled in an IRB-approved, blinded, randomized, controlled trial. Participants were randomized into an intervention (n = 20) and a control (n = 10) group. The intervention group practiced on the FLS suturing task until they achieved expert levels of time and errors (proficiency), were tested on a live porcine fundoplication model, continued simulator training until they achieved expert levels on a visual spatial secondary task (automaticity) and were retested on the operating room (OR) model. The control group participated only during testing sessions. Performance scores were compared within and between groups during testing sessions. RESULTS: : Intervention group participants achieved proficiency after 54 +/- 14 and automaticity after additional 109 +/- 57 repetitions. Participants achieved better scores in the OR after automaticity training [345 (range, 0-537)] compared with after proficiency-based training [220 (range, 0 452; P < 0.001]. CONCLUSIONS: Simulator training to automaticity takes more time but is superior to proficiency-based training, as it leads to improved skill acquisition and transfer. Secondary task metrics that reflect trainee automaticity should be implemented during simulator training to improve learning and skill transfer. PMID- 21637100 TI - Standardized graphs and terms for refractive surgery results. PMID- 21637101 TI - Processes that influence communicative impairments in deaf children using cochlear implants. AB - OBJECTIVES: We previously presented evidence for the existence of a "disproportionate language impairment" (DLI) in a small sample of children using a cochlear implant (CI) who had a good surgical outcome and who were otherwise unimpaired, including typical nonverbal intelligence quotient. Herein, we studied a larger sample of children with DLI who were CI users and, as before, paired them with closely matched, "Control" children who were CI users but had more typical language skills. The primary aim was to assess the communication skills of siblings of each group of CI users to seek evidence for the inheritance of language and more general communicative impairments in the DLI group. We also investigated laterality as a possible coindicator of impairment among the CI users. DESIGN: Prelingually deaf children using a single CI and presenting with severe unexplained language problems (DLI-CI, N = 25) were compared with a group of children using a CI who did not present with such difficulties (Control-CI, N = 25). The groups were matched on age, gender, etiology, age at implantation, implant manufacturer, duration of deafness, and CI experience. Members of both groups completed a series of nonverbal and verbal tests used to identify specific language impairment in normal-hearing (NH) children, and tests of cerebral laterality. Parents completed the Children's Communication Checklist (CCC-2) for each of their children (i.e., the CI users and all their siblings; sibling results were grouped as DLI-Sibling and Control-Sibling) and a questionnaire regarding family history of hearing, speech, language, and literacy problems of the children's first-degree relatives. RESULTS: Significant differences of performance on all language tests were found between the DLI-CI and the Control CI groups. No difference was found between the groups on performance intelligence quotient or auditory memory. CCC-2 results indicated lower General Communication Composite and higher Social Interaction Deviance Composite scores in the DLI-CI than in the Control-CI group. All children in the DLI-CI group produced abnormal overall profiles compared with less than half the children in the Control-CI group. CCC-2 results for the siblings showed a significant association between sibling group and communication; 35% in the DLI-Sibling group produced abnormal profiles compared with 8% in the Control-Sibling group. Co-occurring deafness and language problems in a small number of siblings and other family members did not account fully for these communication difficulties, but there was a higher prevalence of these difficulties in the families of both groups than is reported in NH families. Laterality tasks did not show significant differences between the groups of CI users, although there were trends for children in the DLI-CI group to have less dominant hand preference and eye-hand dominance than those in the Control-CI group and in NH children. CONCLUSIONS: Language tests, including the CCC-2, could identify communication difficulties in children using CIs, leading to prompt identification, informed intervention strategies, and managed expectations. Evidence from their siblings suggests that these difficulties may not all be determined by their deafness or device characteristics but through the same heritable and environmental factors that influence language development in all children. PMID- 21637102 TI - Economic evaluation of targeted cancer interventions: critical review and recommendations. AB - Scientific advances have improved our ability to target cancer interventions to individuals who will benefit most and spare the risks and costs to those who will derive little benefit or even be harmed. Several approaches are currently used for targeting interventions for cancer risk reduction, screening, and treatment, including risk prediction algorithms for identifying high-risk subgroups and diagnostic tests for tumor markers and germline genetic mutations. Economic evaluation can inform decisions about the use of targeted interventions, which may be more costly than traditional strategies. However, assessing the impact of a targeted intervention on costs and health outcomes requires explicit consideration of the method of targeting. In this study, we describe the importance of this principle by reviewing published cost-effectiveness analyses of targeted interventions in breast cancer. Few studies we identified explicitly evaluated the relationships among the method of targeting, the accuracy of the targeting test, and outcomes of the targeted intervention. Those that did found that characteristics of targeting tests had a substantial impact on outcomes. We posit that the method of targeting and the outcomes of a targeted intervention are inextricably linked and recommend that cost-effectiveness analyses of targeted interventions explicitly consider costs and outcomes of the method of targeting. PMID- 21637103 TI - Long-term follow-up of children with confirmed newborn screening disorders using record linkage. AB - BACKGROUND: Long-term follow-up of children identified through newborn screening is a critical process of data collection and analysis for advancing the public health understanding of the health outcomes and service uptake of the affected children. This article describes first steps toward the long-term follow-up of newborn screening children with confirmed disorders through records linkage using population-based administrative data. METHODS: The study cohort consisted of children born in 2006-2007 with confirmed disorders identified through newborn screening. Deterministic data linkage methods were used for record matching. RESULTS: The cohort was followed up to 2 years after birth by matching to data sources including vital records, hospital discharges, the Congenital Malformations Registry, and Early Intervention to monitor service utilization, comorbidities, and mortality of the affected children. Of 1215 children with confirmed conditions identified through newborn screening, 25 deaths (2.1%) were identified, 86.1% used hospital (in- or outpatient) services, 36.1% were enrolled in the Congenital Malformations Registry, and 19.9% used the services of the Early Intervention program during the 2-year follow-up period. CONCLUSIONS: Long term follow-up of children with disorders identified through newborn screening can be initiated by using existing administrative data. This method is an inexpensive, cost-effective. and efficient approach for periodical assessment of services utilization, the efficiency of service delivery, and health outcomes for affected individuals. PMID- 21637104 TI - Why is genetic screening for autosomal dominant disorders underused in families? The case of hereditary hemorrhagic telangiectasia. AB - PURPOSE: Appropriate management of autosomal dominant disorders reduces morbidity and mortality but relies on identifying which family members are affected. Genetic testing may identify relatives needing follow-up but is underused. We conducted this study to identify barriers to genetic testing for one disorder, hereditary hemorrhagic telangiectasia. METHODS: Surveys and online discussion groups with people from hereditary hemorrhagic telangiectasia families. RESULTS: Multiple barriers to hereditary hemorrhagic telangiectasia genetic testing were identified including lack of knowledge about genetic testing, problems with access, and emotional barriers. Many participants did not understand the rationale for hereditary hemorrhagic telangiectasia testing or benefits of early detection; believed that genetic testing is expensive and not covered by insurance; and believed that primary care providers do not know how to order genetic testing. Access to hereditary hemorrhagic telangiectasia testing is limited by distance from a hereditary hemorrhagic telangiectasia center or a genetics clinic. Emotional barriers include fear of insurance discrimination; denial of having hereditary hemorrhagic telangiectasia or being at risk; and guilt and stigma. CONCLUSION: Voluntary disease organizations should develop and disseminate brief educational materials that describe the rationale for genetic testing and emphasize the benefits of early detection and treatment. In addition, laboratories offering genetic testing should provide support for primary care physicians to order and interpret genetic tests. PMID- 21637105 TI - Physicians' communication of Down syndrome screening test results: the influence of physician numeracy. AB - PURPOSE: We investigated three questions: (1) How do obstetrician-gynecologists communicate positive and negative test results? (2) When reporting screening test results, do obstetrician-gynecologists use quantitative or qualitative information? and (3) Is physician numeracy (i.e., the ability to use and understand numbers) associated with use of quantitative or qualitative information? METHOD: Obstetrician-gynecologists (N = 203; 55.6% response rate) who were members of the American College of Obstetricians and Gynecologists completed a survey about their communication of Down syndrome screening test results, an Objective Numeracy Scale, and the Subjective Numeracy Scale. RESULTS: Higher scores on the Subjective Numeracy Scale and younger age predicted obstetrician-gynecologists' use of numbers to explain testing results. The Objective Numeracy Scale did not predict use of numbers. Gender was correlated with scores on the Subjective Numeracy Scale (r = 0.2) and the Subjective Numeracy Scale-Ability Subscale (r = 0.3), with men scoring higher than women when controlling for age. Open-ended questions revealed that communication strategies vary, with approximately one in three obstetrician-gynecologists providing numerical information, and frequency format being the commonly used numerical format. CONCLUSION: Although physicians are often overlooked in the problem of low health literacy, it is important that we continue to investigate the impact of physician numeracy on patient care. PMID- 21637106 TI - COL3A1 haploinsufficiency results in a variety of Ehlers-Danlos syndrome type IV with delayed onset of complications and longer life expectancy. AB - PURPOSE: To characterize the clinical outcome of heterozygosity for COL3A1 null mutations in Ehlers-Danlos syndrome type IV, the vascular type. METHODS: We identified mutations that produced premature termination codons and resulted in nonsense-mediated messenger RNA decay in 19 families. We reviewed the clinical and family histories and medical complications in 54 individuals from these families with COL3A1 null mutations. RESULTS: Compared with individuals with missense or exon-skipping mutations, we found that life span was extended, the age of first complication was delayed by almost 15 years, and major complications were limited to vascular events. The families were ascertained after a complication in a single individual, but only 28% of relatives, some of whom had reached their seventies or eighties without incidents, had a complication and only 30% had minor clinical features of Ehlers-Danlos syndrome type IV CONCLUSION: Null mutations have reduced penetrance compared with missense and splicing mutations, and the phenotype seems to be limited almost entirely to vascular events. PMID- 21637107 TI - The impact of antibodies on clinical outcomes in diseases treated with therapeutic protein: lessons learned from infantile Pompe disease. AB - PURPOSE: Enzyme replacement therapy with rhGAA (Myozyme(r)) has lead to improved survival, which is largely attributable to improvements in cardiomyopathy and skeletal muscle function. However, crossreactive immunologic material-negative patients have a poor clinical response to enzyme replacement therapy secondary to high sustained antibody titers. Furthermore, although the majority of crossreactive immunologic material-positive patients tolerize or experience a downtrend in anti-rhGAA antibody titers, antibody response is variable with some crossreactive immunologic material-positive infants also mounting high sustained antibody titers. METHODS: We retrospectively analyzed 34 infants with Pompe disease: 11 crossreactive immunologic material-negative patients, nine high-titer crossreactive immunologic material-positive patients, and 14 low-titer crossreactive immunologic material-positive patients. Clinical outcome measures were overall survival, ventilator-free survival, left ventricular mass index, Alberta Infant Motor Scale score, and urine Glc(4) levels. RESULTS: Clinical outcomes in the high-titer crossreactive immunologic material-positive group were poor across all areas evaluated relative to the low-titer crossreactive immunologic material-positive group. For the crossreactive immunologic material negative and high-titer crossreactive immunologic material-positive groups, no statistically significant differences were observed for any outcome measures, and both patient groups did poorly. CONCLUSIONS: Our data indicate that, irrespective of crossreactive immunologic material status, patients with infantile Pompe disease with high sustained antibody titer have an attenuated therapeutic response to enzyme replacement therapy. With the advent of immunomodulation therapies, identification of patients at risk for developing high sustained antibody titer is critical. PMID- 21637108 TI - Unsafe sex among HIV-infected adults in Kenya: results of a nationally representative survey. AB - OBJECTIVE: Assess factors associated with knowledge of HIV status, sexual activity, and unprotected sex with a partner of unknown or negative HIV status (unsafe sex) among HIV-infected adults in Kenya. DESIGN: Nationally representative Kenya AIDS Indicator Survey among adults aged 15-64 years in 2007. METHODS: A standardized questionnaire was administered and blood samples tested for HIV. We assessed factors associated with knowledge of HIV infection, sexual activity, and unsafe sex. Analyses took into account stratification and clustering in the survey design and estimates were weighted to account for sampling probability. RESULTS: Of 15,853 participants with blood samples, 1104 (6.9%) were HIV infected. Of these, 83.8% did not know their HIV status (56% had never tested; 27.8% reported their last HIV test was negative), and 80.4% were sexually active. Of 861 sexually active adults, 76.9% reported unsafe sex in the past year. Adults who did not know their HIV status were more likely to be sexually active [never tested adjusted odds ratio (AOR): 5.5, 95% confidence interval (CI): 2.8 to 10.7; ever tested, incorrect knowledge AOR: 6.5, CI: 2.1 to 19.6) and to report unsafe sex (never tested AOR: 51.7, CI: 27.3 to 97.6; ever tested, incorrect knowledge of status AOR: 18.6, CI: 8.6 to 40.5) than those who knew their status. CONCLUSIONS: The majority of adults did not know they were infected and engaged in unsafe sex. Adults who knew their HIV status were less likely to be sexually active and report unsafe sex compared with those unaware of their infection. HIV prevention interventions that target HIV-infected adults are urgently needed. PMID- 21637109 TI - Interleukin-2 production by polyfunctional HIV-1-specific CD8 T cells is associated with enhanced viral suppression. AB - BACKGROUND: Assays to measure the induction of HIV-1-specific CD8 T-cell responses often rely on measurements of indirect effector function such as chemokine and cytokine production, which may not reflect direct elimination of an invading pathogen. Assessment of the functional ability of CD8 T cells to suppress HIV-1 replication has been viewed as a surrogate marker of an effectual immune response. To further investigate this, we measured the capacity of virus specific CD8 T cells to inhibit HIV-1 replication in an in vitro suppression assay. METHODS: We expanded 15 epitope-specific CD8 T-cell lines from peripheral blood mononuclear cells of chronically HIV--infected progressors (n = 5) and controllers (n = 4) who were not on antiretroviral therapy. Cell lines were tested for their ability to produce effector molecules (CD107a, IL-2, IFN-gamma, TNF-alpha, perforin) and suppress virus replication in autologous CD4 T cells. RESULTS: CD8 T-cell lines from both progressors and controllers had largely similar effector function profiles as determined by intracellular cytokine staining. In contrast, we observed that CD8 T-cell lines derived from controllers show enhanced virus suppression when compared with progressors. Virus suppression was mediated in an major histocompatibility complex-dependent manner and found to correlate with a polyfunctional IL-2 CD8 T-cell response. CONCLUSIONS: Using a sensitive in vitro suppression assay, we demonstrate that CD8 T-cell-mediated suppression of HIV-1 replication is a marker of HIV-1 control. Suppressive capacity was found to correlate with polyfunctional IL-2 production. Assessment of CD8 T-cell-mediated suppression may be an important tool to evaluate vaccine induced responses. PMID- 21637111 TI - Validation in Uganda of the new WHO diagnostic algorithm for smear-negative pulmonary tuberculosis in HIV prevalent settings. AB - BACKGROUND: We compared the effectiveness of the new WHO 2007 diagnostic algorithm for pulmonary tuberculosis (PTB) for HIV prevalent settings (WHO07) to the WHO 2003 guidelines used by the Ugandan National Tuberculosis Program (UgWHO03) and most other countries for the diagnosis of smear-negative PTB. METHODS: A prospective observational cohort design was employed under routine operational conditions at a District Hospital in Uganda. Newly diagnosed patients with HIV and enrolled to the outpatient HIV clinic were assessed for possible PTB. The study was in 2 phases with evaluation of the UgWHO03 and WHO07 over the first and second phases, respectively. RESULTS: One hundred forty-seven enrolled onto the UgWHO03 and 166 onto WHO07. There were significant reductions in the geometric mean days (11.0 vs. 21.2, P < 0.05) and number of health facility visits (1.7 vs. 2.8, P < 0.001) from presentation to diagnosis of all PTB for the WHO07 compared with the UgWHO03. For smear-negative PTB, the geometric mean days (28.5 vs. 34.1, P > 0.05) and number of health facility visits (2.5 vs. 2.6, P > 0.05) from presentation to diagnosis were nonsignificantly reduced. Diagnostic sensitivity (95% confidence intervals) for smear-negative PTB increased nonsignificantly from 63% (27% to 90%) to 95% (72% to 100%) for the WHO07, with specificity (95% confidence interval) remaining high at 99% (92% to 100%) and 98% (93% to 100%). CONCLUSIONS: Implementing the new WHO07 is likely to reduce the number of days and increase sensitivity for diagnosis of smear-negative cases of PTB. We identified many barriers to implementing both guidelines in HIV prevalent resource-poor settings. PMID- 21637110 TI - Augmented HIV-specific interferon-gamma responses, but impaired lymphoproliferation during interruption of antiretroviral treatment initiated in primary HIV infection. AB - BACKGROUND: Antiretroviral therapy (ART) introduced during primary HIV infection followed by treatment interruption (TI) is postulated to enhance virologic control through induction of HIV-specific CD4 T cells, which foster virus specific CD8+ T cells that suppress virus replication. This hypothesis was evaluated in 21 subjects enrolled in AIDS Clinical Trials Group 709, a substudy of AIDS Clinical Trials Group 371, which prospectively evaluated subjects who received >=1 year of ART initiated in acute or recent HIV infection followed by TI. METHODS: Lymphoproliferation was assessed by [methyl-H] thymidine incorporation and HIV-specific CD8+ T-cell interferon-gamma responses by enzyme linked immunospot-forming assays. Virologic success was defined as sustained viral load <5000 copies per milliliter for 24 weeks after TI. RESULTS: HIV specific lymphoproliferative responses were detected at least once in 5 (24%) of 21 subjects, were generally transient, and were unrelated to HIV-specific interferon-gamma responses (P > 0.4). HIV-specific CD8+ interferon-gamma responses increased after 48 weeks of ART (P = 0.03), but failed to predict virologic success (P = 0.18). Compared with seronegative subjects, lymphoproliferation to Candida, cytomegalovirus, and alloantigens was similar in HIV-infected subjects during ART, but lower during TI (P <= 0.04). CONCLUSIONS: HIV-specific CD8+ T-cell interferon-gamma responses expand during ART following primary HIV infection, but are not related to HIV-specific lymphoproliferative responses nor virologic success. Impaired non-HIV antigen-specific lymphoproliferation associated with TI suggests this strategy could be deleterious. PMID- 21637112 TI - Effect of mutations at position E138 in HIV-1 reverse transcriptase on phenotypic susceptibility and virologic response to etravirine. AB - The contribution of E138 mutations to etravirine resistance was investigated. Amino acids at position E138 after failure with etravirine in DUET were A (n = 1), G (n = 5), K (n = 3), P (n = 1), Q (n = 5), and V (n = 2). At baseline, only E138A and Q were found at 3.0% and 2.5%, respectively. Virologic response (less than 50 copies/mL) was observed in six of 12 and eight of 10 patients with E138A and E138Q, respectively. Site-directed mutants harboring E138A/G/K/Q/R or S showed etravirine fold change values of 2.9, 2.4, 2.6, 3.0, 3.6, and 2.8, respectively. E138G, K, and Q were added to the existing etravirine-weighted genotypic score including 17 etravirine resistance-associated mutations. PMID- 21637113 TI - Delayed maturation of antibody avidity but not seroconversion in rhesus macaques infected with simian HIV during oral pre-exposure prophylaxis. AB - BACKGROUND: Pre-exposure prophylaxis (PrEP) is a novel intervention strategy for the prevention HIV transmission. Because several clinical trials are at various stages of completion, it is important to understand the impact of PrEP treatment on the development of the immune response to HIV, particularly in individuals who exhibit breakthrough infections despite PrEP. METHODS: A model of HIV infection, using rhesus macaques and the simian/human immunodeficiency virus (SHIV), was used to evaluate the effects of PrEP on the evolution of the humoral immune response. Time to seroconversion, neutralizing and binding antibody levels, and antibody avidity were measured in 12 rhesus macaques infected during daily or intermittent PrEP with FTC (emtricitabine) or Truvada (FTC/tenofovir combination) and compared with 11 untreated, simian HIV-infected controls. RESULTS: Macaques that became infected while receiving PrEP exhibited significantly lower peak virus loads during acute infection as compared with untreated animals. Although the timing of seroconversion and SHIV binding and neutralizing antibody levels were not impacted by treatment, lower maturation rates of antibody avidity for anti-p27, gp120, gp160, and gp41 were observed. CONCLUSIONS: This study suggests that reduced virus loads associated with PrEP treatment have little impact on timing of seroconversion and neutralizing/binding antibody levels; however, maturation of antibody avidity was suppressed. PMID- 21637114 TI - Disparities in antiretroviral treatment: a comparison of behaviorally HIV infected youth and adults in the HIV Research Network. AB - OBJECTIVES: Increasing numbers of youth are becoming HIV-infected and need highly active antiretroviral therapy (HAART). We hypothesized that behaviorally HIV infected youth (BIY) ages 18 to 24 years are less likely than adults (25 years or older) to receive HAART and, once initiated, more likely to discontinue their first HAART regimen. METHODS: Longitudinal analysis of treatment-naive patients (age 18 years or older) meeting criteria for HAART and followed at HIV Research Network sites (2002-2008). Time from meeting criteria to HAART initiation and duration on first regimen were assessed using Cox proportional hazards regression. RESULTS: A total of 3127 (268 youth, 2859 adult) treatment-naive, HIV infected patients met criteria. BIY were more likely to be black (66.8% vs 51.1%; P < 0.01) and less likely to identify injection drug use HIV risk (1.1% vs 8.8%; P < 0.01) than adults 25 years of age or older. Nearly 69% of BIY started HAART versus 79% of adults (P < 0.001). Adults 25 to 29 years of age (adjusted hazards ratio [AHR], 1.39; 95% confidence interval [CI], 1.12-1.73) and 50 years of age or older (AHR, 1.24; 95% CI, 1.00-1.54), but not 30 to 49 years (AHR, 1.19; 95% CI, 0.99-1.44) were more likely to initiate HAART than BIY. Attending four or more HIV provider visits within 1 year of meeting criteria was associated with HAART initiation (AHR, 1.91; 1.70-2.14). CD4 200 to 350 versus less than 200 cells/mm (AHR, 0.57; 95% CI, 0.52-0.63), and injection drug use (AHR, 0.80; 95% CI, 0.69-0.92) were associated with a lower likelihood of HAART initiation. There were no age-related differences in duration of the first regimen. CONCLUSION: BIY are less likely to start HAART when meeting treatment criteria. Addressing factors associated with this disparity is critical to improving care for youth. PMID- 21637115 TI - Glaucoma diagnostic capabilities of optic nerve head parameters as determined by Cirrus HD optical coherence tomography. AB - PURPOSE: To compare the glaucoma diagnostic capabilities of optic nerve head (ONH) parameters with retinal nerve fiber layer thickness (RNFLT) using Cirrus spectral-domain optical coherence tomography (Carl Zeiss Meditec Inc, Dublin, CA; version 5.0.0.326). METHODS: Two hundred twenty-nine glaucomatous patients, 405 preperimetric glaucoma patients, and 109 healthy individuals were imaged by Cirrus optical coherence tomography optic disc cube mode. Correlations were sought between RNFLT and ONH parameters (disc and rim area, average and vertical cup-to-disc ratio, and cup volume). Areas under receiver operating characteristic curves (AUCs) of average RNFLT were compared with those of ONH parameters with respect to discrimination between glaucomatous patients and healthy individuals. Subgroup analysis was performed in early, moderate-to-advanced glaucomatous groups, glaucoma patients with a small disc area and a large disc area. RESULTS: Rim area showed the strongest correlation with average RNFLT (r=0.663) and the highest AUC (0.871). The overall AUC for discrimination between healthy individuals and glaucomatous patients was higher for average RNFLT than for rim area (0.957 vs. 0.871, P<0.001). In the early and small disc area subgroup, the AUC of average RNFLT was significantly greater than those of all ONH parameters. In moderate-to-advanced glaucomatous groups' patients, the AUCs of average RNFLT and rim area, in large disc area group patients, the AUC of average RNFLT and vertical cup-to-disc ratio, did not differ significantly. CONCLUSIONS: RNFLT was better than any tested ONH parameter when used for glaucoma discrimination, especially in patients with early-stage glaucoma and in glaucomatous patients with small optic discs. PMID- 21637116 TI - Comparison of retinal nerve fiber layer measurement between 2 spectral domain OCT instruments. AB - PURPOSE: To determine the repeatability of retinal nerve fiber layer (RNFL) measurement from 2 spectral domain optical coherence tomography (OCT) instruments and assess the level of agreement. A secondary objective was to assess if pupillary dilatation affected the repeatability of RNFL measurement. DESIGN: Prospective, observational study. PARTICIPANTS: Fifty normal participants were recruited after being screened for exclusion criteria, which included preexisting glaucoma and significant cataract. METHODS: Peripapillary RNFL thickness was measured from either eye of the 50 participants using both the Cirrus and Spectralis OCTs. Three scans were performed consecutively for each instrument and the process was repeated after pupillary dilatation. MAIN OUTCOME MEASURES: Test retest variability, coefficient of variation, and intraclass correlation coefficient (ICC) for repeatability of RNFL measurement. ICC and Bland-Altman plots for agreement between both devices. RESULTS: Test-retest variability for the Cirrus and Spectralis OCTs were 4.89 MUm and 4.95 MUm, respectively. RNFL measurements were less repeatable in the 4 quadrants, ranging from 5.02 to 15.00 MUm. The temporal quadrant was the most repeatable quadrant for the Cirrus OCT with a variability of 7.81 MUm, whereas the nasal quadrant was the most repeatable for Spectralis OCT with 5.02 MUm. Test-retest variability was poorer for the Cirrus OCT in the nasal, inferior, and superior quadrants. Pupillary dilatation increased repeatability of RNFL measurement for Cirrus OCT, particularly in the superior quadrant, whereas not affecting measurements from Spectralis OCT. ICC values for agreement between both instruments were good. For average RNFL as well as RNFL in the superior, inferior, and temporal quadrants, ICC ranged from 0.859 to 0.908. The nasal quadrant had a lower ICC of 0.663. Bland-Altman plots showed that the Spectralis OCT provided the thicker RNFL measurement in general, except in the nasal quadrant when RNFL was thin. CONCLUSIONS: The repeatability of RNFL thickness measurement in normal participants was excellent for both the Cirrus and Spectralis OCTs. Agreement of RNFL measurement between both the devices was generally good, with the exception of the nasal quadrant in which a linear relationship exists. Pupillary dilatation improved the repeatability of RNFL measurement for Cirrus while having minimal influence on Spectralis OCT. More studies will be required to ascertain the relationships of RNFL measurement between the different spectral domain OCT instruments in normal and glaucomatous patients. PMID- 21637117 TI - Posterolateral approaches for the treatment of tibial plateau fractures and total knee arthroplasty. PMID- 21637118 TI - Fat emboli syndrome in a nondisplaced tibia fracture. PMID- 21637119 TI - How does a varus deformity of the humeral head affect elevation forces and shoulder function? A biomechanical study with human shoulder specimens. AB - OBJECTIVES: A biomechanical study was performed to test the hypothesis that a varus deformity of the humeral head decreases supraspinatus (SSP) efficiency and increases deltoid elevation forces in human specimens. METHODS: Twenty-four fresh frozen human shoulder specimens were prepared by preserving the rotator cuff and deltoid. A defined, medial closed-wedge osteotomy was performed and lateral locked plate applied to simulate a varus deformity of 45 degrees in Group I (n = 8) and 20 degrees in Group II (n = 8). The control group (n = 8) was not osteotomized. The effect of the deformities on arm elevation forces was measured in a robot-assisted shoulder simulator under a physiologically loaded rotator cuff during three elevation phases. Phase 1 encompassed 0 degrees to 30 degrees , Phase 2 was from 30 degrees to 60 degrees , and Phase 3 included 60 degrees to 90 degrees of shoulder elevation. RESULTS: SSP efficiency, defined as the degree of elevation attained per unit muscle force, was significantly less in Group I compared with Group II (P = 0.036) and the control group (P = 0.039) (Group I = 0.12 +/- 0.03 degrees /N, Group II 0.18 +/- 0.05 degrees /N, and control group 0.24 +/- 0.10 degrees /N). Under physiological loading of the rotator cuff, the deltoid (DELT) elevation forces were significantly greater in Group I (Pphase 1 = 0.015, Pphase 3 = 0.001) and Group II (Pphase 1 = 0.015, Pphase 3 = 0.006) compared with the control group in elevation Phase 1 (Group I: 3.20 +/- 1.04 N/ degrees , Group II: 3.03 +/- 0.96 N/ degrees , control group: 2.01 +/- 0.53 N/ degrees ) and Phase 3 (Group I: 2.50 +/- 0.85 N/ degrees , Group II: 1.55 +/- 0.28 N/ degrees , control group: 1.21 +/- 0.18 N/ degrees ). When the SSP was unloaded, the DELT elevation forces were significantly greater in Group l than in Group II (P = 0.040) and the control group (P = 0.004) during elevation Phase 3 (Group I: 2.12 +/- 0.60 N/ degrees , Group II: 1.47 +/- 0.34 N/ degrees , control group: 1.24 +/- 0.32 N/ degrees ). CONCLUSIONS: A varus deformity of the humeral head changes the pretension of the rotator cuff and results in a significantly decreased SSP efficiency (45 degrees varus) and significantly higher arm elevation forces (20 degrees varus). Clinically, the study's findings are relevant because they indicate that varus deformities of more than 20 degrees should not be accepted intraoperatively and might indicate the need for surgical correction in case of subsequent symptoms. PMID- 21637120 TI - Biomechanical evaluation of two-part surgical neck fractures of the humerus fixed by an angular stable locked intramedullary nail. AB - OBJECTIVE: The aim of the current study was to see how different interlocking mechanisms would affect construct stability and overall failure in the treatment of two-part surgical neck fractures in the proximal humerus in vitro. METHODS: Left and right bones of eight pairs of fresh-frozen human cadaveric humeri were assigned to either a group with conventional or a group with angular stable distal interlocking. The different experimental interlocking mechanisms were used in a surgical neck fracture model of the humerus (Orthopaedic Trauma Association 11- A3) stabilized by a proximal humeral nail. The following variables were evaluated by biomechanical tests: hysteresis width in bending and torsion, stiffness, and fracture gap movement during cyclic axial loading until failure and the overall failure mechanism of the construct. RESULTS: The angular stable group showed significantly less motion in initial bending and torsion and higher bending stiffness throughout the complete deformation cycle compared with the conventional interlocked group. Fracture gap movement was significantly less in the angular stable group. Higher stability was mainly observed in the early phase of the applied loading pattern; however, ultimate failure was not related to distal interlocking but occurred in the proximal fragment in both groups. CONCLUSIONS: An experimental angular stable distal interlocking system of proximal humeral nails shows higher construct stability in the early phase of fracture fixation in vitro. In terms of overall failure, loss of fixation in the proximal fragment was crucial and not different between groups. PMID- 21637121 TI - Reconstructive surgical treatment without bone grafting in nonunions of humeral surgical neck fractures. AB - OBJECTIVE: To evaluate reconstruction surgery without bone grafting in humeral surgical neck nonunions using either a blade plate or the "Humerusblock" implant in small head fragments. DESIGN: Retrospective cohort study, Evidence-Based Medicine Level IV. SETTING: Paracelsus Medical University Salzburg, Department of Traumatology and Sports Injuries. PATIENTS: Fifty-five patients (mean age, 66.2 years) with symptomatic nonunion after humeral surgical neck fractures were subjected to surgical reconstruction. INTERVENTION: In 45 patients, open reduction and internal fixation using a blade plate was performed (Group 1). In 10 patients, the Humerusblock device was used (Group 2), because the head fragment of each of these patients was too small for plating. In no patients was bone grafting used. MAIN OUTCOME MEASUREMENTS: Shoulder function, level of pain, and bone healing. RESULTS: After a mean follow-up time of 74 months, the overall mean Constant score improved from 30.4% preoperatively to 83.2% postoperatively. The improvement was from 30.5% to 85.3% in Group 1 and from 32.2% to 75.4% in Group 2, which represented statistically significant improvements for both groups (P < 0.01). Radiologic bone healing was achieved in 51 patients (93%). The overall complication rate was high, 15%, and complications included plate loosening, avascular head necrosis, persistent nonunion, and infection. CONCLUSIONS: Nonunion of humeral surgical neck fractures can be successfully treated by surgical reconstruction without bone grafting using either a blade plate or the Humerusblock for small head fragments. However, an increased complication rate is associated with this challenging posttraumatic pathology. PMID- 21637122 TI - A fragment-specific approach to Type IID Monteggia elbow fracture-dislocations. AB - OBJECTIVES: To describe the pattern of injury, surgical technique, and outcomes of Monteggia Type IID fracture dislocations. DESIGN: Retrospective review of prospectively collected clinical and radiographic patient data in an orthopaedic trauma database. SETTING: Level I university-based trauma center. PATIENTS/PARTICIPANTS: All patients with Monteggia Type IID fracture-dislocations admitted from January 2000 to July 2005. INTERVENTION: Review of patient demographics, fracture pattern, method of fixation, complications, additional surgical procedures, and clinical and radiographic outcome measures. MAIN OUTCOME MEASUREMENTS: Clinical outcomes: elbow range of motion, complications. Radiographic outcomes: characteristic fracture fragments, quality of fracture reduction, healing time, degenerative changes, and heterotopic ossification. RESULTS: Sixteen patients were included in the study. All fractures united. There were six complications in six patients, including three contractures with associated heterotopic ossification, one pronator syndrome and late radial nerve palsy, one radial head collapse, and one with prominent hardware. CONCLUSIONS: Monteggia IID fracture-dislocations are complex injuries with typical specific fracture fragments. Anatomic fixation of all injury components and avoidance of complications where possible can lead to a good outcome in these challenging injuries. PMID- 21637123 TI - Hang them high: a hands-free technique for limb-holding during surgical preparation. AB - We describe the use of a suspension system to provide hands-free elevation and traction of lower limbs during preoperative skin preparation. The technique is particularly useful for fractured limbs that are often both awkward to elevate by hand and present a physical challenge to lift and hold. Furthermore, limb-lifting and holding has been identified by the National Association of Orthopaedic Nurses as a commonly performed task associated with high risk of work-related musculoskeletal disorders. The National Association of Orthopaedic Nurses Task Force has recommended the use of assisted lifting and holding devices if the task of limb elevation and or holding exceeds its recommended limits. The principal determinants of these limits are the weight of the leg, the height it is elevated above the chest, and the time for which it is held. The maximal weight recommended for a two hand lift is 35 pounds, which is often less than the weight of a patient's leg whose total body weight is over 190 pounds. The maximal limit is below 35 pounds for lifts above chest height and for holds over 1 minute. Because the average size and weight of American citizens is increasing, so has the need for devices to assist with lifting and holding in the operating room. This technique also can improve operating room efficiency because a member of the operating team is not engaged in holding the limb during the skin preparation and draping. The limb is also held reliably with constant in-line traction by this technique. This reduces the chance for accidental decontamination of the skin or further movement of the fracture and potential secondary soft tissue damage if the limb holder loses or changes their grip. PMID- 21637125 TI - Calculating the mounting parameters for Taylor Spatial Frame correction using computed tomography. AB - The Taylor Spatial Frame uses a computer program-based six-axis deformity analysis. However, there is often a residual deformity after the initial correction, especially in deformities with a rotational component. This problem can be resolved by recalculating the parameters and inputting all new deformity and mounting parameters. However, this may necessitate repeated x-rays and delay treatment. We believe that error in the mounting parameters is the main reason for most residual deformities. To prevent these problems, we describe a new calculation technique for determining the mounting parameters that uses computed tomography. This technique is especially advantageous for deformities with a rotational component. Using this technique, exact calculation of the mounting parameters is possible and the residual deformity and number of repeated x-rays can be minimized. This new technique is an alternative method to accurately calculating the mounting parameters. PMID- 21637124 TI - Decreasing bacterial colonization of external fixation pins through nitric oxide release coatings. AB - OBJECTIVE: Bacterial infection of the pin tract represents the most common complication associated with external fixation. This study was designed to evaluate the antibacterial activity of nitric oxide (NO)-releasing xerogel films applied to commercially pure titanium pins in a rat model. METHODS: Pins were coated with xerogel solution through a dip-coating procedure. Half of the xerogel coated implant pins were modified into NO donors and served as the NO-releasing group, whereas the remaining pins were left unmodified to serve as non-NO releasing xerogel-coated controls. Acid-etched pins served as uncoated controls. Animal selection was randomized and every rat had one pin from each of the three groups randomly allocated to the third, fourth, or fifth tail vertebrae. Quantification of bacterial infection was performed 48 days postoperatively and the tissue-implant interface was inspected for clinical signs of infection on Days 14 and 28 postimplantation. RESULTS: Pin tract bacterial colony counts of the NO-releasing group (170,000 +/- 181,000) were significantly lower than both the xerogel-coated group (677,000 +/- 675,000) and the control group (1,181,000 +/- 2,717,000) 48 days postoperatively (P < 0.05). No significant difference in colony counts was observed between the xerogel-coated group and the control group. The NO-releasing group also had significantly fewer clinical signs of infection than both the coated and the control groups on postoperative Day 28 (P < 0.05). CONCLUSION: The application of NO-releasing xerogel coatings can inhibit bacterial colonization of external fixation pins both during the initial postsurgical period and up to 48 days postimplantation. PMID- 21637126 TI - The pin-bone interface in external fixator: a standardized analysis in a sheep osteotomy model. AB - OBJECTIVES: The hypothesis of the study was that the incidence of pin loosening and pin infection would increase, whereas the general stability of the pin-bone interface would decrease with ongoing implantation time. The aim of this study was to analyze the biologic reactions of the bone tissue adjacent to the pin to determine the relationship among the osseous anchorage of pins, the incidence of infections, and the histologic appearance. METHODS: Three groups of sheep received a tibial osteotomy stabilized by external fixators. The pin-bone interface was analyzed biomechanically, radiologically, microbiologically, and histologically after 3, 6, and 9 weeks. RESULTS/CONCLUSIONS: Contrary to common opinion, pin anchorage was not altered biomechanically throughout the 9 weeks of the study. This effect might be attributed to an increasing remodeling found in the callus and cortex around the pins and was likely assisted by a strict pin care routine and a low infection rate. PMID- 21637128 TI - Interaction among ethnicity, socioeconomic status, and Helicobacter pylori seroprevalence in Israeli children and adolescents. AB - OBJECTIVES: The aim of the study was to examine the prevalence of Helicobacter pylori infection among Israeli children from different backgrounds and to assess potential interactions between ethnicity, socioeconomic status (SES), and H pylori seroprevalence. PATIENTS AND METHODS: The present sero-epidemiologic study was conducted among 0- to 20-year-old children seeking medical attention, not specifically gastrointestinal symptoms, using sera collected between 2000 and 2001 from 575 Israeli Arab children, 584 Jewish children from the general population, and sera that were obtained between 1997 and 2007 from 464 children of an ultraorthodox Jewish community. An enzyme-linked immunosorbent assay was used to measure H pylori serum immunoglobulin G antibodies and seropositivity to H pylori CagA strains. RESULTS: H pylori seropositivity was 22.9% (95% confidence interval [CI] 19.7-26.5) among Jewish children from the general population, 25.2% (95% CI 21.5-29.4) among ultraorthodox Jewish children, and 45.6% (95% CI 41.5 49.7) among Arab children. H pylori seroprevalence increased significantly with age in the 3 study groups, but it was consistently higher in Arab children. Compared with Jewish participants from high SES and controlling for age and sex, the odds ratio for H pylori seropositivity was 2.03 (95% CI 1.31-3.12) in Jewish children from intermediate SES, 2.42 (95% CI 1.29-4.53) in Arab children from intermediate SES, 2.26 (95% CI 1.52-3.36) in Jewish children from low SES, and 5.72 (95% CI 3.89-8.42) in Arab children from low SES. CagA seropositivity was 40.8% and 45.0% among Jewish and Arab children, respectively (P =0.59), and it was highest among subjects of lower SES. CONCLUSIONS: Socioeconomic factors may not totally explain the ethnic differences in H pylori prevalence. PMID- 21637127 TI - Vitamin D deficiency is prevalent in girls and women with Rett syndrome. AB - OBJECTIVES: The aim of the study was to determine the prevalence of vitamin D deficiency and identify the relation between 25-hydroxyvitamin D (25-(OH)D) levels and the consumption of dietary sources of vitamin D or exposure to anticonvulsants in girls and women with Rett syndrome (RTT). SUBJECTS AND METHODS: Retrospective review of the medical records of 284 girls and women with RTT to determine serum 25-(OH)D and parathyroid hormone levels, nutritional status, dietary sources of vitamin D, exposure to anticonvulsants, degree of mobility, and MECP2 status. RESULTS: Twenty percent of girls and women who were tested (n = 157) had 25-(OH)D levels <50 nmol/L. Multivitamin supplements, vitamin D-fortified milk, and commercial formulas were consumed by 40%, 52%, and 54%, respectively. Anticonvulsants were used by 57%, and 39% ambulated independently. Median 25-(OH)D levels were lower in individuals who did not receive multivitamin supplements (P < 0.05) or commercial formulas (P < 0.001) than in those who did. Median 25-(OH)D levels differed (P < 0.01) among racial and ethnic groups, but the number in some groups was small. Nutritional status, use of anticonvulsants, degree of mobility, and MECP2 status did not influence 25 (OH)D levels. CONCLUSIONS: Vitamin D deficiency is prevalent in girls and women with RTT. The use of multivitamin supplements or commercial formulas is associated with improved vitamin D levels. Attention to vitamin D may enhance bone mineral deposition and reduce the frequency of bone fractures in these individuals. PMID- 21637129 TI - Hamartomatous small intestinal polyp from tuberous sclerosis. PMID- 21637130 TI - Rising incidence of inflammatory bowel disease in young children: What does the future hold? PMID- 21637131 TI - Efficacy of an MFGM-enriched complementary food in diarrhea, anemia, and micronutrient status in infants. AB - OBJECTIVE: The aim of the present study was to evaluate the efficacy of a milkfat globule membrane (MFGM)-enriched protein fraction in a complementary food, on diarrhea, anemia, and micronutrient status. SUBJECTS AND METHODS: A randomized, double-blind controlled design to study 550 infants, 6 to 11 months old, who received daily for 6 months a complementary food (40 g/day) with the protein source being either the MFGM protein fraction or skim milk proteins (control). Health and nutritional status of infants were examined monthly in the outpatient clinic; product intake, food patterns, and diarrhea morbidity were assessed by home visits twice per week. Hemoglobin and micronutrient status were measured at 0 and 6 months of intervention. Results are presented as the entire group and as 6 to 8 and 9 to 11 months subgroups. RESULTS: A total of 499 infants completed the study. Global prevalence of diarrhea was 3.84% and 4.37% in the MFGM group and control group, respectively (P < 0.05). Consumption of the MFGM protein fraction reduced episodes of bloody diarrhea (odds ratio 0.54; 95% confidence interval 0.31-0.93, P = 0.025) adjusting for anemia and potable water facilities as covariates. There were no differences between groups in anemia, serum ferritin, zinc, or folate. CONCLUSIONS: Addition of an MFGM-enriched protein fraction to complementary food had beneficial effects on diarrhea in infants and may thus help to improve the health of vulnerable populations. PMID- 21637132 TI - Claustrophobia: a proxy for psychological distress in patients with back pain. AB - STUDY DESIGN: Case-control study. OBJECTIVE: The aim of this study was to assess the levels of psychological distress in patients with back pain who expressed claustrophobia at the time of their magnetic resonance imaging (MRI) scan, compared with sex and age-matched normal controls who did not exhibit claustrophobia. The secondary aim was to document the level of disability and intervention rates in this group. SUMMARY OF BACKGROUND DATA: Psychosocial factors influence the outcomes of low back pain treatment with psychological distress being associated with poorer surgical outcomes in patients with low back pain. Up to 14% of patients experience claustrophobia during MRI scans requiring sedation to complete the scan. The effect of claustrophobia on back pain disability and outcomes has not been previously reported. METHODS: Twenty females and 13 males all requiring MRI scan under sedation for claustrophobia (group 1) were compared with an age and sex-matched cohort that had MRI scan without sedation (group 2). Both groups were drawn from a chronic back clinic. Average age in both groups was 54 years (range, 27 to 79 y). Both groups had standard conservative therapy, together with psychometric evaluation: Zung Depression Index and Modified Somatic Perception Questionnaire. Disability was measured by Oswestry Disability Index. Primary outcome measures were intervention rates (surgery, injections, and physiotherapy sessions) and prevalence of psychological distress. RESULTS: Mean Zung Depression Index in group 1 was significantly higher than in group 2 (59.5 vs. 28.9, P<0.05) as was the mean Modified Somatic Perception Questionnaire score (13.3 vs. 9.2, P<0.05.) Prevalence of psychological distress was higher in group 1 (75.8% vs. 18.2%, P<0.05). Oswestry Disability Index was the same in both groups (50% vs. 48%). Group 1 had 13 interventions (13 patients=39.4%) compared with 26 in group 2 (26 patients=78.8%) (P< 0.05). Twenty-two patients (66.7%) in group 1 were discharged after their MRI scan with no intervention compared with 7 patients (21.2%) in group 2 (P<0.05). CONCLUSIONS: Claustrophobic patients with back pain showed higher levels of depression than nonclaustrophobic patients, with a higher rate of psychological distress. Disability, however, was not higher. The majority of claustrophobic patients did not require surgical intervention. The reasons for this are unclear and require further investigation. Claustrophobia requiring sedation for MRI scans may be a proxy for psychological distress in these patients and psychometric testing is advised during assessment to help with surgical decision making. PMID- 21637133 TI - Spinopelvic dissociation as a complication of iliac crest bone graft harvest using an acetabular reamer. AB - STUDY DESIGN: Descriptive case report. OBJECTIVE: To report spinopelvic dissociation as a complication of autogenous bone graft harvest with the use of an acetabular reamer. SUMMARY OF BACKGROUND DATA: Iliac crest bone graft harvest has been previously described in orthopedic literature with overall complication rates of 0% to 39%. Complications are inconsistently separated into "minor" and "major" groups, commonly reporting hematoma, infection, and injury to the superior gluteal artery or nerve. No reports of spinopelvic dissociation, a well known injury in the traumatic setting, have been reported. METHODS: We report the case of 1 patient who presented with spinopelvic dissociation after autogenous bone graft harvest with the use of an acetabular reamer. RESULTS: We present a unique case in which fracture through the reamer harvesting site led to complete disruption of the posterior pelvic ring on the operative side. Subsequent attempted closed management led to failure through the osteoporotic anterior pelvic ring on the operative side and the posterior pelvic ring on the contralateral side, leading to progressive instability, disability, and pain. This was successfully addressed with lumbopelvic fusion and 2-year follow-up is presented. CONCLUSIONS: The potential hazard of acetabular reamer bone graft harvest is discussed, and primary operative fixation of harvest-related fractures is suggested. PMID- 21637134 TI - Anterior Approach Versus Posterior Approach With Subtotal Corpectomy, Decompression, and Reconstruction of Spine in the Treatment of Thoracolumbar Burst Fractures: A Prospective Randomized Controlled Study. AB - STUDY DESIGN: A randomized, controlled follow-up study. OBJECTIVE: The objective of this study was to compare the results of anterior approach versus posterior approach with subtotal corpectomy, decompression, and reconstruction of spine in the treatment of thoracolumbar burst fractures. SUMMARY OF BACKGROUND DATA: Burst fractures are frequently associated with instability or neurological deficit. Anterior subtotal corpectomy, decompression, and reconstruction with instrumentation are an established method for a highly unstable burst fracture. In the past few years, subtotal corpectomy, decompression, and reconstruction of spine could be completed by posterior approach. Posterior segmental pedicle screw instrumentation, with its more rigid fixation and less technically demanding, could offer potential advantages. METHODS: A total of 64 patients with thoracolumbar burst fractures were divided into 2 groups randomly. Group A was treated by anterior approach and group B was treated by posterior approach with subtotal corpectomy, decompression, and reconstruction of spine. During the minimum 24 months (range, 24 to 72 mo) follow-up period, all patients were prospectively evaluated for clinical and radiologic outcomes. The intraoperative blood loss, operative time, complications of operation, pulmonary function, Frankel scale, and the American Spinal Injury Association (ASIA) motor score were used for clinical evaluation, whereas the heights of anterior edge of vertebral body and the Cobb angle were examined for radiologic outcome. RESULTS: All patients in this study achieved solid fusion, with significant neurological improvement. The intraoperative blood loss (P<0.05) and complications of operation were less, the operative time was shorter (P<0.05), and the pulmonary function after operation was better in the group B (P<0.05). The Frankel scale, the ASIA motor score, and the radiologic results were not significantly different (P<0.05) at all time points between the 2 groups A and B. But the 2 groups improved in their neurological function by approximately 1.3 Frankel grade and 15.6 ASIA motor scores at final follow-up. CONCLUSION: Anterior approach and posterior approach with subtotal corpectomy, decompression, and reconstruction of spine are sufficient for surgical treatment of thoracolumbar burst fractures. Less intraoperative blood loss and complications, shorter operative time, and better pulmonary function after operation are the significant advantages of posterior surgery. PMID- 21637135 TI - Reconstruction of extensive defects from posterior en bloc resection of sacral tumors with human acellular dermal matrix and gluteus maximus myocutaneous flaps. AB - BACKGROUND: Performing a sacrectomy from an exclusively posterior approach allows the en bloc resection of tumors without the morbidity of a laparotomy. However, reconstruction of the resultant extensive soft-tissue defects is challenging because a vertical rectus abdominis myocutaneous flap is not harvested. OBJECTIVE: To report the largest series (with the longest follow-up) of sacral reconstructions using a combination of human acellular dermal matrix (HADM) and gluteus maximus myocutaneous flaps. METHODS: Thirty-four patients with sacral tumors with a follow-up of at least 1 year were reviewed retrospectively. After the tumor was excised, HADM (AlloDerm, LifeCell Corp, Branchburg, New Jersey) was secured to create a pelvic diaphragm. Subsequently, the gluteus maximus muscles were freed from their origins and advanced to cover the HADM. RESULTS: The mean age of patients was 50.1 years (SD, 16.0 years), and the histopathology was a chordoma in 82.4%. Seven patients (20.6%) developed a postoperative wound dehiscence, 5 of whom (14.7%) required operative debridement. An estimated blood loss of >1500 mL, an operative time of >9 hours during sacrectomy, and postoperative bowel incontinence were associated with a significantly higher likelihood of undergoing a subsequent debridement for dehiscence (P <= .03). With a mean follow-up of 45.7 months, only 1 patient developed an asymptomatic parasacral hernia. CONCLUSION: Reconstruction of posterior sacrectomy defects with HADM and gluteus maximus myocutaneous flaps may be valid. This approach may have rates of wound dehiscence comparable to other techniques and low rates of parasacral herniation. PMID- 21637136 TI - Challenges in the management of ruptured and unruptured brainstem arteriovenous malformations: outcome after conservative, single-modality, or multimodality treatments. AB - BACKGROUND: Brainstem arteriovenous malformations are challenging lesions, and benefits of treatment are uncertain. OBJECTIVE: To study the clinical course of Brainstem arteriovenous malformations and the influence of treatments on outcome. METHODS: We reviewed a prospective series of 31 brainstem arteriovenous malformations. Demographic, morphological, and clinical characteristics were recorded. Factors determining initial and final outcomes (modified Rankin Scale), results of treatments (cure rates, complications), and disease course were analyzed. RESULTS: Brainstem arteriovenous malformations were symptomatic and bled in 93% and 61% of cases, respectively. Examination was abnormal and initial modified Rankin Scale score was < 3 in 71% and 86% of patients, respectively. The average follow-up time was 6.2 years, and 26% of patients rebled (5.9 %/y). Treatment modalities included conservative, radiosurgical, endovascular, surgical, and multimodality treatment in 13%, 58%, 35%, 16%, and 26% of cases, respectively. The obliteration rate was 60% overall and 39% after radiosurgery, 40% after embolization, and 75% after microsurgery, with respective complication free cure rates of 71%, 50%, and 0%. Overall procedural mortality and morbidity were 2.3% and 18.6%, respectively. Final modified Rankin Scale score was < 3 in 77% of cases. Neurological deterioration (35%) was related to treatment complications in 74% of cases with a negative impact of surgery (P = .04), palliative embolization (odds ratio = 16), and multimodality treatments (odds ratio = 24). Radiosurgery was inversely associated with worsening (odds ratio = 0.06). CONCLUSION: Brainstem arteriovenous malformations require individualized treatment decisions. Single-modality treatments with a reasonable chance of complete cure and low complication rate (such as radiosurgery) should be favored. PMID- 21637137 TI - The benefit of subthalamic deep brain stimulation for pain in Parkinson disease: a 2-year follow-up study. AB - BACKGROUND: Pain is a well-recognized feature of Parkinson disease (PD), which is primarily a motor disorder. In a previous study, we showed that subthalamic deep brain stimulation (STN DBS) improves pain as well as motor symptoms 3 months after surgery in PD patients. OBJECTIVE: To determine whether there is a long term beneficial effect of STN DBS on pain in PD. METHODS: We studied 21 patients with PD who underwent STN DBS. Motor symptoms were assessed using the Unified Parkinson's Disease Rating Scale and Hoehn and Yahr staging. Pain was evaluated by asking patients about the quality and severity of pain in each body part. Evaluations were performed at baseline and at 3 and 24 months after surgery. RESULTS: At baseline, 18 of the 21 patients (86%) experienced pain. After surgery, most of the pain reported at baseline had improved or disappeared at 3 months and improved further at 24 months. The benefit of STN DBS for pain evaluated at 24 months was comparable to that with medication at baseline. At 24 months, 9 patients (43%) reported new pain that was not present at baseline. Most of the new pain was musculoskeletal in quality. Despite the development of new pain, the mean pain score at follow-up was lower than at baseline. CONCLUSION: STN DBS improves pain in PD, and this beneficial effect persists, being observed after a prolonged follow-up of 24 months. In addition, in many of the PD patients new, mainly musculoskeletal pain developed on longer follow-up. PMID- 21637138 TI - Adenosine for temporary flow arrest during intracranial aneurysm surgery: a single-center retrospective review. AB - BACKGROUND: Clip application for temporary occlusion is not always practical or feasible. Adenosine is an alternative that provides brief periods of flow arrest that can be used to advantage in aneurysm surgery, but little has been published on its utility for this indication. OBJECTIVE: To report our 2-year consecutive experience with 40 aneurysms in 40 patients for whom we used adenosine to achieve temporary arterial occlusion during aneurysm surgery. METHODS: We retrospectively reviewed our clinical database between May 2007 and December 2009. All patients who underwent microsurgical clipping of intracranial aneurysms under adenosine induced asystole were included. Aneurysm characteristics, reasons for adenosine use, postoperative angiographic and clinical outcome, cardiac complications, and long-term neurological follow-up with the modified Rankin Scale were noted. RESULTS: Adenosine was used for 40 aneurysms (10 ruptured, 30 unruptured). The most common indications for adenosine were aneurysm softening in 17 cases and paraclinoid location in 14 cases, followed by broad neck in 12 cases and intraoperative rupture in 6 cases. Troponins were elevated postoperatively in 2 patients. Echocardiography did not show acute changes in either. Clinically insignificant cardiac arrhythmias were noted in 5 patients. Thirty-six patients were available for follow-up. Mean follow-up was 12.8 months. The modified Rankin Scale score was 0 for 29 patients at the time of the last follow-up. Four patients had an modified Rankin Scale score of 1, and scores of 2 and 3 were found in 2 and 1 patients, respectively. CONCLUSION: Adenosine appears to allow safe flow arrest during intracranial aneurysm surgery. This can enhance the feasibility and safety of clipping in select circumstances. PMID- 21637139 TI - Antibody-mediated rejection after alemtuzumab induction: incidence, risk factors, and predictors of poor outcome. AB - BACKGROUND: Antibody-mediated rejection (AMR) is associated with allograft loss. Identification of factors associated with poor outcome has not been extensively studied. METHODS: We retrospectively studied 469 patients who received a negative crossmatch renal transplant with alemtuzumab induction. Forty-eight of 469 (10.2%) patients were treated for AMR. Thirty of 48 (62.5%) of the cases fulfilled the Banff criteria for definite AMR, whereas 18 of 48 (37.5%) were categorized as suspicious for AMR (tissue injury with C4d staining or donor specific antibodies [DSAbs]). Sensitization, high human leukocyte antigen, and DR mismatch were risk factors for the development of AMR (P = 0.0016, 0.001, and 0.012, respectively). RESULTS: Allograft survival was inferior in the AMR group (70.2%) compared with the nonrejector group (97.0%) (P<0.001). Forty-two of 48 (87.5%) of patients with acute AMR had DSAbs. Patients with CII DSAbs at the time of AMR, whether alone or in combination with CI DSAbs had the worst allograft survival (P = 0.014). Both the mean cumulative and immunodominant mean fluorescence index were higher in those patients who subsequently lost their grafts (P<0.001). Patients with diffuse C4d staining had inferior allograft survival than those with focal C4d or no staining (P = 0.02). There was no significant difference in survival by histological grade but a trend to inferior outcomes in those with vascular involvement (P = 0.06). Those patients who met the full Banff criteria had worse survival than those with suspicion for AMR only (P = 0.04). CONCLUSION: This study identifies patients at risk of graft failure from AMR. These patients may benefit from newer therapeutic strategies including the use of eculizumab or bortezomib. PMID- 21637140 TI - Pretransplant IgG subclasses of donor-specific human leukocyte antigen antibodies and development of antibody-mediated rejection. AB - BACKGROUND: The subclass of IgG antibodies contributes to their capability to activate complement. It is currently unknown whether the pretransplant IgG subclass composition allows distinguishing harmful from presumably irrelevant donor-specific human leukocyte antigen (HLA) antibodies (HLA-DSA) detected by single-antigen flow beads (SAFB). METHODS: Seventy-four patients transplanted in the presence of HLA-DSA were investigated. HLA-DSA characteristics were not different between patients experiencing antibody-mediated rejection (AMR) (n=40) and patients who did not (n=34) experience AMR. Sera were reanalyzed using SAFB with IgG subclass-specific reporter antibodies. RESULTS: The 74 patients had in total 141 HLA-DSA. IgG1 was the predominant subclass (78%), followed by IgG2 (49%), IgG3 (36%), and IgG4 (20%). When grouped according to the complement activating capability, only 4 of 74 patients (5%) had exclusively weak/no complement-activating HLA-DSA (i.e., IgG2 and IgG4), 21 of 74 patients (28%) had isolated strong complement-activating HLA-DSA (i.e., IgG1 and IgG3), and 46 of 74 patients (62%) had a mixture of both. There was no difference between the strong complement-activating and the mixture group regarding incidence of AMR (57% vs. 54%; P=0.81), phenotypes of AMR (P=0.70), and death-censored allograft survival at 5 years (78% vs. 78%; P=0.74). Interestingly, patients with exclusively weak/no complement-activating HLA-DSA (n=4) had a numerically lower incidence of AMR (25%) and no allograft loss has occurred yet. CONCLUSION: In 90% of patients, pretransplant HLA-DSA are composed of isolated strong or a mixture of strong and weak/no complement-activating IgG subclasses. Because outcomes in these two groups were similar, pretransplant IgG subclass analysis is likely not providing substantial value beyond the standard IgG SAFB assay for pretransplant risk stratification. PMID- 21637141 TI - Moving forward: advancing venous thromboembolism knowledge in pediatric trauma. PMID- 21637142 TI - Liver support for acute liver failure: plus ca change, plus c'est la meme chose. PMID- 21637143 TI - Glucose control in the pediatric intensive care unit: more questions than answers! PMID- 21637144 TI - Checklists, guidelines, and protocols: a prescription for improved outcomes. PMID- 21637145 TI - Infections acquired while on extracorporeal membrane oxygenation: navigating the maze. PMID- 21637146 TI - Antibiotics in the pediatric intensive care unit: have we finally figured out that less is more? PMID- 21637147 TI - Ventilator-associated infections: beat the regulators to the punch: a call to action. PMID- 21637148 TI - Where in the world is WALDO (Why Adrenal Levels Defy Observation)? Hemodynamic correlates of serum cortisol in neonates after cardiopulmonary bypass. PMID- 21637149 TI - Quality and safety in the pediatric cardiac intensive care unit: it is time to leave the pit stop and the cockpit and perfect the handover. PMID- 21637150 TI - Predicting outcome after severe pediatric traumatic brain injury: making progress one baby step at a time. PMID- 21637151 TI - Bereavement in the neonatal period: what we know and what we wish. PMID- 21637152 TI - Strategic approaches to improving arrest management: the promise of technology and the limits of advance training. PMID- 21637153 TI - Prolonged mechanical ventilation: does shorter duration of mechanical ventilation equal morbidity reduction for congenital heart disease patients? PMID- 21637154 TI - Therapeutic hypothermia: ready...fire...aim? How small feasibility studies can inform large efficacy trials. PMID- 21637155 TI - Planning for the next influenza pandemic: what should we expect? PMID- 21637156 TI - How much practice makes perfect? PMID- 21637157 TI - A call for full public disclosure for donation after circulatory determination of death in children. PMID- 21637159 TI - Comparative proteomic analysis of irinotecan-sensitive colorectal carcinoma cell line and its chemoresistant counterpart. AB - In this study, we used two-dimensional gel electrophoresis and MALDI-Q-TOF-MS/MS analysis to examine the global protein expression of a pair of colorectal carcinoma cell lines, SW620 and irinotecan-resistant SW620. Of the 30 spots identified as differentially expressed proteins (+/-over twofold, P<0.05) between the two cell lines, 26 spots (corresponding to 26 unique proteins) were positively identified by MALDI-Q-TOF-MS/MS analysis. These proteins could be grouped into main classes including metabolism (15.38%), cell SSproliferation/differentiation (11.53%), molecular chaperone (11.53%), mRNA splicing (11.53%), and so on. The proteins, which might be involved in the development of tumor drug resistance, such as alpha-enolase, cofilin, and thioredoxin-dependent peroxide 1, have been validated by western blot analysis and have been discussed. The proteins identified in this study may be useful in showing the mechanisms underlying irinotecan resistance. PMID- 21637160 TI - Cytotoxic effects of the trifunctional bispecific antibody FBTA05 in ex-vivo cells of chronic lymphocytic leukaemia depend on immune-mediated mechanism. AB - Monoclonal antibodies such as rituximab and alemtuzumab show considerable therapeutic efficacy in chronic lymphocytic leukaemia (CLL). Aiming to further improve antineoplastic efficacy, the trifunctional bispecific antibody FBTA05 was developed. FBTA05 is thought to function by simultaneously binding B cells and T cells by its variable regions and by recruiting FcgammaR-positive accessory immune cells by its intact Fc region. As it was previously shown that this antibody shows considerable cytotoxicity towards a spectrum of B-cell lymphoma cell lines, we here tested its potential efficacy ex vivo against malignant B-CLL cells. Therefore, we assessed the capacity of increasing concentrations of FBTA05 to bind to neoplastic cells, to induce cytotoxicity (comparing it with rituximab and alemtuzumab) and cytokine release. We evaluated the results with respect to the extent of CD20 expression, the effector:target cell ratio as well as with the patients' overall effector cell status. Thus, we show that, although FBTA05 elicited cytotoxicity was comparable with that induced by alemtuzumab, it considerably exceeded the antineoplastic effects of rituximab. Noteworthy, FBTA05 shows effective elimination of malignant B cells even if CD20 surface expression is low. Importantly, a high grade of cytotoxicity was associated with the induction of T-cell proliferation and the concomittant release of interferon gamma and interleukin-6, thus overcoming the detrimental effects of an unfavourable effector:target cell ratio. In conclusion, we here present novel evidence for the therapeutic efficacy of the trifunctional, bispecific antibody FBTA05 in CLL and provide evidence for the importance of immune-mediated mechanisms conveying the cytotoxic effects against malignant B lymphocytes. PMID- 21637161 TI - An in-vitro evaluation of the polo-like kinase inhibitor GW843682X against paediatric malignancies. AB - Polo-like kinase 1 (PLK1) is a regulator of mitosis and its upregulation in tumours is often associated with poor prognosis. Although PLK1 inhibitors have already entered phase 1 clinical trials, little is known about their impact on the treatment of paediatric malignancies. Thus, we evaluated the concept of PKL1 inhibition by testing the effects of the PLK1 inhibitor GW843682X alone and in combination with the topoisomerase 1 inhibitor, camptothecin, against a panel of 18 paediatric tumour cell lines. Cytotoxicity was evaluated by MTT test and by caspase 3/7 activation. Expression of target was confirmed by western blot analysis. Expression of ATP binding cassette transporters was analysed by quantitative real-time reverse transcription PCR. GW843682X significantly inhibited cell growth in all 18 cell lines. Concentrations, which inhibited cell growth by 50% compared with untreated controls after 72 h, ranged from 0.02 to 11.7 MUmol/l. Apart from the N-Myc-amplified neuroblastoma cell lines, the osteosarcoma cell lines MNNG-HOS and OST, which are highly resistant to standard anticancer drugs, were sensitive to GW843682X. The toxicity of GW843682X was dependent neither on the ATP binding cassette drug transporter expression nor on the p53 mutation status. Neither synergistic nor antagonistic effects were observed for the combination of GW843682X and camptothecin in 14 cell lines. GW843682X showed considerable toxicity against a panel of paediatric tumour cell lines suggesting that PLK1 inhibitors under clinical development should be evaluated against paediatric malignancies too. PMID- 21637162 TI - Adenovirus-mediated p53 gene therapy reverses resistance of breast cancer cells to adriamycin. AB - The aim of this study was to determine whether adenovirus-mediated p53 gene (Ad p53) transfection can enhance adriamycin cytotoxicity and reverse adriamycin resistance in human breast cancer cells and explore its effect on the expression of MDR1 gene and permeability-glycoprotein (P-gp). Human breast cancer cell lines, MCF-7 and MCF-7/ADR, were used in in-vitro studies. After infection with Ad-p53, the cytotoxicity of adriamycin was evaluated using the Cell Counting Kit 8 assay. The expression of MDR1 mRNA was detected by quantitative real-time PCR. The expression of P-gp was analyzed using western blotting. In in-vivo studies, MCF-7/ADR tumor cells were inoculated subcutaneously in athymic nude mice. After 14 days of inoculation, tumor size was measured. Apoptosis and expression of P-gp in the tumor tissue were analyzed by fluorescence activated cell sorting and western blotting. After transfection with a multiplicity of infection of 50 for Ad-p53, chemosensitivity of MCF-7/ADR cells increased by 18.1 times (P=0.001), and 50% inhibitory concentration (IC50) of adriamycin decreased from 4.54 +/- 0.91 to 0.26 +/- 0.11 mg/l. Real-time PCR showed that MDR1 mRNA decreased from 1.32 to 0.85 (P=0.001). Western blotting analysis showed that P-gp also decreased. In in-vivo studies, Ad-p53 combined with adriamycin dramatically inhibited the growth of subcutaneous xenograft of MCF-7/ADR. The fluorescence activated cell sorting assay showed that there were more apoptotic cells in tumor tissues treated with Ad-p53 and adriamycin. The expression of P-gp was significantly decreased in tumor tissues. This study suggests that Ad-p53 can reverse MCF-7/MDR cell resistance to adriamycin. The reversal effect was associated with inhibition of P-gp expression and induction of apoptosis. PMID- 21637164 TI - Bibliography. Obstetric and gynaecological anesthesia. Current world literature. PMID- 21637165 TI - Counting down to good health. 10 tips from the editors. PMID- 21637166 TI - Menopausal hot flashes tied to lower breast cancer risk. PMID- 21637167 TI - Studies offer more support for use of shingles vaccine. PMID- 21637168 TI - Know your numbers. Key figures offer insight into your health. PMID- 21637169 TI - Mayo Clinic office visit. Solid fats and added sugars (SoFAS). An interview with Jennifer Nelson, M.S., R.D. PMID- 21637170 TI - Cultivating contentment. Turn up your happiness level. PMID- 21637171 TI - Is Kombucha tea safe to drink? PMID- 21637172 TI - Is mouth-to-mouth breathing necessary for CPR to be effective? PMID- 21637173 TI - Special report. Understanding arthritis. A look at common conditions, treatment options. PMID- 21637174 TI - Update from the New Jersey Department of Banking and Insurance. PMID- 21637175 TI - Perspectives on healthcare from the 2011 Edward J. Ill Excellence in Medicine honorees. PMID- 21637176 TI - Profile of Mary O'Dowd, MPH. Acting Commissioner, New Jesey Department of Health and Senior Services. PMID- 21637177 TI - Prescribing in an age of "wonder drugs". PMID- 21637178 TI - Pathologizing old age: the case of osteoporosis. PMID- 21637179 TI - New legislation to regulate out-of-network benefits. PMID- 21637180 TI - Repairing the communication disconnect. PMID- 21637181 TI - Medically unexplained symptoms in the veteran population: challenges and opportunities. PMID- 21637182 TI - HIV surveillance--United States, 1981-2008. AB - Within 1 year of the initial report in 1981 of a deadly new disease that occurred predominantly in previously healthy persons and was manifested by Pneumocystis carinii pneumonia and Kaposi's sarcoma, the disease had a name: acquired immune deficiency syndrome (AIDS). Within 2 years, the causative agent had been identified: human immunodeficiency virus (HIV). On the 30th anniversary of the epidemic, to characterize trends in HIV infection and AIDS in the United States during 1981-2008, CDC analyzed data from the National HIV Surveillance System. This report summarizes the results of that analysis, which indicated that, in the first 14 years, sharp increases were reported in the number of new AIDS diagnoses and deaths among persons aged>=13 years, reaching highs of 75,457 in 1992 and 50,628 in 1995, respectively. With introduction of highly active antiretroviral therapy, AIDS diagnoses and deaths declined substantially from 1995 to 1998 and remained stable from 1999 to 2008 at an average of 38,279 AIDS diagnoses and 17,489 deaths per year, respectively. Despite the decline in AIDS cases and deaths, at the end of 2008 an estimated 1,178,350 persons were living with HIV, including 236,400 (20.1%) whose infection was undiagnosed. These findings underscore the importance of the National HIV/AIDS Strategy focus on reducing HIV risk behaviors, increasing opportunities for routine testing, and enhancing use of care (1). PMID- 21637183 TI - HIV testing among men who have sex with men--21 cities, United States, 2008. AB - Although men who have sex with men (MSM) comprise an estimated 2% of the overall U.S. population aged>=13 years (1), 59% of persons with diagnoses of human immunodeficiency virus (HIV) infection in the United States in 2009 were MSM, including MSM who inject drugs (2). CDC recommends HIV testing at least annually for sexually active MSM to identify HIV infections and prevent ongoing transmission (3). Results of HIV testing conducted as part of the National HIV Behavioral Surveillance System (NHBS) in 21 cities indicated that 19% of MSM who were tested in 2008 were HIV-positive; of these, 44% were unaware that they were infected (4). To assess whether MSM were tested as recommended and whether more frequent testing might be indicated, CDC analyzed NHBS data for 2008. This report describes the results of that analysis, which indicated that, of 7,271 MSM interviewed and tested who did not report a previous positive HIV test, 61% had been tested for HIV infection during the past 12 months; among these, 7% had a new, positive HIV test result when tested as part of NHBS. Given the high prevalence of new HIV infection among MSM who had been tested during the past year, sexually active MSM might benefit from more frequent HIV testing (e.g., every 3 to 6 months). PMID- 21637184 TI - Vaccination coverage among children in kindergarten--United States, 2009-10 school year. AB - Healthy People 2020 objectives include maintaining vaccination coverage among children in kindergarten (IID-10) (1). The target is >=95% vaccination coverage for the following vaccines: poliovirus; diphtheria and tetanus toxoids and acellular pertussis (DTP/DTaP/DT); measles, mumps, and rubella (MMR); hepatitis B (HepB); and varicella (1). Data from school assessment surveys are used to monitor vaccination coverage and vaccination exemption levels among children enrolled in kindergarten. This report summarizes data from school assessment surveys submitted to CDC by 48 federal immunization program grantees (including 47 states and the District of Columbia) for the 2009-10 school year to describe vaccination coverage and exemption rates (2). For that period, 17 grantees reported coverage of ?95% for four vaccines (poliovirus, DTP/DTaP/DT, MMR, and HepB) and four grantees reported coverage of >=95% for 2 doses of varicella vaccine. Total exemption rates, including medical, religious, and philosophical exemptions, ranged from <1% to 6.2% across grantees, and 15 grantees reported exemption rates<1%. Survey methods for vaccination coverage and exemption rates varied among grantees, making comparisons difficult and limiting the use of school assessment surveys to report aggregate national rates. Further standardization of school assessment survey methods will generate comparable data between grantees to monitor and track progress in reaching national objectives, and allow development of best practice guidelines for grantees to more effectively use and report school coverage and exemption data. CDC will continue to monitor vaccination coverage and exemption levels and assist grantees in identification of local areas with low vaccination coverage or high exemption rates for further evaluation or intervention. PMID- 21637185 TI - Update: influenza activity--United States, 2010-11 season, and composition of the 2011-12 influenza vaccine. AB - During the 2010-11 influenza season, influenza activity* first began to increase in the southeastern United States, and peaked nationally in early February. Compared with the previous pandemic year (2009-10), higher rates of hospitalization were observed for persons aged>=65 years during the 2010-11 season, whereas lower hospitalization rates were observed in younger populations than during the pandemic year. Overall, the percentages of outpatient visits for influenza-like illness (ILI) were lower during the 2010-11 season than the 2009 10 pandemic influenza season. In the United States, influenza A (H3N2) remained the predominant virus throughout the season; however, 2009 influenza A (H1N1) and influenza B viruses also circulated, and the predominant virus varied by U.S. Department of Health and Human Service (HHS) region and week. This report summarizes influenza activity in the United States during the 2010-11 influenza season (October 3, 2010-May 21, 2011) and describes the components of the 2011-12 Northern Hemisphere influenza vaccine. PMID- 21637192 TI - Lower back-up rates improve ventilator triggering during assist-control ventilation: a randomized crossover trial. AB - OBJECTIVE: The objective of this study is to compare the effects of back-up ventilation rates (BURs) on triggered inflations and patient cardiorespiratory stability during assist-control/volume guarantee ventilation (AC/VG). STUDY DESIGN: This study is a randomized crossover trial conducted in a neonatal unit in an Australian tertiary NICU. In all, 26 stable preterm infants on AC/VG ventilation were studied at BUR settings of 30, 40 and 50 min(-1). Inflation rate, triggering and cardiorespiratory measures of patient stability were compared during 20 min epochs with 10 min washout periods. RESULT: The 26 infants studied were median (inter-quartile range) gestational age 27 (26, 30) weeks, birth weight 0.84 (0.75, 1.14) kg and FiO(2) 0.24 (0.21, 0.31) and age 6 (4, 19) days. At BURs of 30, 40 and 50, the proportions of inflations, which were triggered, were mean (s.d.) 85% (11), 75% (19) and 61% (25); P<0.01 for all comparisons. Total delivered inflation rates were 56 (8), 58 (9) and 62 (8) min( 1), respectively. Cardiorespiratory parameters did not vary between the settings. CONCLUSION: Using a lower BUR allows greater triggering of ventilator inflations. Cardiorespiratory parameters including CO(2) levels were stable at all rates. PMID- 21637193 TI - Faster generation of hiPSCs by coupling high-titer lentivirus and column-based positive selection. AB - The protocols described here address methods used in two crucial stages in the retroviral reprogramming of somatic cells to produce human induced pluripotent stem cell (hiPSC) lines. The first is an optimized method for producing lentivirus at an efficiency 600-fold greater than previously published, and it includes conjugation of the lentivirus to streptavidin superparamagnetic particles; this process takes 8 d. The second method enables the isolation of true hiPSCs immediately after somatic cell reprogramming and involves column based positive selection of cells expressing the pluripotency marker TRA-1-81. This process takes 2 h and, as it is directly compatible with feeder-free culture, the time burden of manually identifying and mechanically propagating hiPSC colonies is reduced drastically. Taken together, these methods accelerate the production of hiPSCs and enable lines to be isolated, expanded to approxiamtely 107 cells and cryopreserved within 6-8 weeks. PMID- 21637194 TI - Initiation, growth and cryopreservation of plant cell suspension cultures. AB - Methods described in this paper are confined to in vitro dedifferentiated plant cell suspension cultures, which are convenient for the large-scale production of fine chemicals in bioreactors and for the study of cellular and molecular processes, as they offer the advantages of a simplified model system for the study of plants when compared with plants themselves or differentiated plant tissue cultures. The commonly used methods of initiation of a callus from a plant and subsequent steps from callus to cell suspension culture are presented in the protocol. This is followed by three different techniques for subculturing (by weighing cells, pipetting and pouring cell suspension) and four methods for growth measurement (fresh- and dry-weight cells, dissimilation curve and cell volume after sedimentation). The advantages and disadvantages of the methods are discussed. Finally, we provide a two-step (controlled rate) freezing technique also known as the slow (equilibrium) freezing method for long-term storage, which has been applied successfully to a wide range of plant cell suspension cultures. PMID- 21637195 TI - Web-based inference of biological patterns, functions and pathways from metabolomic data using MetaboAnalyst. AB - MetaboAnalyst is an integrated web-based platform for comprehensive analysis of quantitative metabolomic data. It is designed to be used by biologists (with little or no background in statistics) to perform a variety of complex metabolomic data analysis tasks. These include data processing, data normalization, statistical analysis and high-level functional interpretation. This protocol provides a step-wise description on how to format and upload data to MetaboAnalyst, how to process and normalize data, how to identify significant features and patterns through univariate and multivariate statistical methods and, finally, how to use metabolite set enrichment analysis and metabolic pathway analysis to help elucidate possible biological mechanisms. The complete protocol can be executed in approximately 45 min. PMID- 21637196 TI - Synthesis of all-hydrocarbon stapled alpha-helical peptides by ring-closing olefin metathesis. AB - This protocol provides a detailed procedure for the preparation of stapled alpha helical peptides, which have proven their potential as useful molecular probes and as next-generation therapeutics. Two crucial features of this protocol are (i) the construction of peptide substrates containing hindered alpha-methyl, alpha-alkenyl amino acids and (ii) the ring-closing olefin metathesis (RCM) of the resulting resin-bound peptide substrates. The stapling systems described in this protocol, namely bridging one or two turns of an alpha-helix, are highly adaptable to most peptide sequences, resulting in favorable RCM kinetics, helix stabilization and promotion of cellular uptake. PMID- 21637197 TI - Rapid DNA detection by beacon-assisted detection amplification. AB - This protocol describes a new and rapid isothermal reaction process designed to amplify and detect a specific DNA sequence in purified DNA extracted from cultured cells. The protocol uses a DNA nanomachine that comprises two molecular switches that function in concert to isothermally amplify and detect a DNA target. First, a molecular beacon detection switch is 'activated' only if a DNA target sequence is present. A DNA primer and DNA polymerase are used to lock the beacon in an activated conformation. Second, an amplification and signal transduction switch is initiated following successful activation. A nicking endonuclease and the DNA polymerase are used to replicate the DNA target. Both switches operate simultaneously at 40 degrees C in a single reaction to rapidly generate multiple copies of the DNA target in a cyclic polymerization reaction. This protocol enables femtomole amounts of a DNA target to be reproducibly amplified and detected in <40 min. We demonstrate the successful use of this protocol in assays containing synthetic DNA components and purified DNA extracted from biological samples. PMID- 21637198 TI - Flexizymes for genetic code reprogramming. AB - Genetic code reprogramming is a method for the reassignment of arbitrary codons from proteinogenic amino acids to nonproteinogenic ones; thus, specific sequences of nonstandard peptides can be ribosomally expressed according to their mRNA templates. Here we describe a protocol that facilitates genetic code reprogramming using flexizymes integrated with a custom-made in vitro translation apparatus, referred to as the flexible in vitro translation (FIT) system. Flexizymes are flexible tRNA acylation ribozymes that enable the preparation of a diverse array of nonproteinogenic acyl-tRNAs. These acyl-tRNAs read vacant codons created in the FIT system, yielding the desired nonstandard peptides with diverse exotic structures, such as N-methyl amino acids, D-amino acids and physiologically stable macrocyclic scaffolds. The facility of the protocol allows a wide variety of applications in the synthesis of new classes of nonstandard peptides with biological functions. Preparation of flexizymes and tRNA used for genetic code reprogramming, optimization of flexizyme reaction conditions and expression of nonstandard peptides using the FIT system can be completed by one person in approximately 1 week. However, once the flexizymes and tRNAs are in hand and reaction conditions are fixed, synthesis of acyl-tRNAs and peptide expression is generally completed in 1 d, and alteration of a peptide sequence can be achieved by simply changing the corresponding mRNA template. PMID- 21637199 TI - Generation of phospholipid vesicle-nanotube networks and transport of molecules therein. AB - We describe micromanipulation and microinjection procedures for the fabrication of soft-matter networks consisting of lipid bilayer nanotubes and surface immobilized vesicles. These biomimetic membrane systems feature unique structural flexibility and expandability and, unlike solid-state microfluidic and nanofluidic devices prepared by top-down fabrication, they allow network designs with dynamic control over individual containers and interconnecting conduits. The fabrication is founded on self-assembly of phospholipid molecules, followed by micromanipulation operations, such as membrane electroporation and microinjection, to effect shape transformations of the membrane and create a series of interconnected compartments. Size and geometry of the network can be chosen according to its desired function. Membrane composition is controlled mainly during the self-assembly step, whereas the interior contents of individual containers is defined through a sequence of microneedle injections. Networks cannot be fabricated with other currently available methods of giant unilamellar vesicle preparation (large unilamellar vesicle fusion or electroformation). Described in detail are also three transport modes, which are suitable for moving water-soluble or membrane-bound small molecules, polymers, DNA, proteins and nanoparticles within the networks. The fabrication protocol requires ~90 min, provided all necessary preparations are made in advance. The transport studies require an additional 60-120 min, depending on the transport regime. PMID- 21637200 TI - A simple, versatile and efficient method to genetically modify human monocyte derived dendritic cells with HIV-1-derived lentiviral vectors. AB - Lentiviral vectors derived from the human immunodeficiency type 1 virus (HIV-1 LV) are among the finest tools available today for the genetic modification of human monocyte-derived dendritic cells (MDDCs). However, this process is largely inefficient because MDDCs show a strong resistance to HIV-1 transduction. Here we describe a step-by-step protocol from the production of LVs to cell transduction that allows the efficient genetic modification of MDDCs. This protocol can be completed in 23 d from the initial phase of LV production to the final analysis of the results of MDDC transduction. The method relies on the simultaneous addition of HIV-1 LVs along with noninfectious virion-like particles carrying Vpx, a nonstructural protein encoded by the simian immunodeficiency virus (Vpx VLPs). When thus provided in target cells, Vpx exerts a strong positive effect on incoming LVs by counteracting the restriction present in MDDCs; accordingly, 100% of cells can be transduced with low viral inputs. Vpx-VLPs will improve the efficiency of LV-mediated transduction of MDDCs with vectors for both ectopic gene expression and depletion studies. PMID- 21637201 TI - Mouse corneal lymphangiogenesis model. AB - This protocol describes a powerful in vivo method to quantitatively study the formation of new lymphatic vessels in the avascular cornea without interference of pre-existing lymphatics. Implantation of 100 ng of lymphangiogenic factors such as vascular endothelial growth factor (VEGF)-A, VEGF-C or fibroblast growth factor-2, together with slow-release polymers, into a surgically created micropocket in the mouse cornea elicits a robust lymphangiogenic response. Newly formed lymphatic vessels are detected by immunohistochemical staining of the flattened corneal tissue with lymphatic endothelial-specific markers such as lymphatic vessel endothelial hyaluronan receptor-1; less-specific markers such as vascular endothelial growth factor receptor 3 may also be used. Lymphatic vessel growth in relation to hemangiogenesis can be readily detected starting at day 5 or 6 after pellet implantation and persists for ~14 d. This protocol offers a unique opportunity to study the mechanisms underlying lymphatic vessel formation, remodeling and function. PMID- 21637202 TI - Generating gene knockout rats by homologous recombination in embryonic stem cells. AB - We describe here a detailed protocol for generating gene knockout rats by homologous recombination in embryonic stem (ES) cells. This protocol comprises the following procedures: derivation and expansion of rat ES cells, construction of gene-targeting vectors, generation of gene-targeted rat ES cells and, finally, production of gene-targeted rats. The major differences between this protocol and the classical mouse gene-targeting protocol include ES cell culture methods, drug selection scheme, colony picking and screening strategies. This ES cell-based gene-targeting technique allows sophisticated genetic modifications to be performed in the rat, as many laboratories have been doing in the mouse for the past two decades. Recently we used this protocol to generate Tp53 (also known as p53) gene knockout rats. The entire process requires ~1 year to complete, from derivation of ES cells to generation of knockout rats. PMID- 21637203 TI - Imaging three-dimensional tissue architectures by focused ion beam scanning electron microscopy. AB - In this protocol, we describe a 3D imaging technique known as 'volume electron microscopy' or 'focused ion beam scanning electron microscopy (FIB/SEM)' applied to biological tissues. A scanning electron microscope equipped with a focused gallium ion beam, used to sequentially mill away the sample surface, and a backscattered electron (BSE) detector, used to image the milled surfaces, generates a large series of images that can be combined into a 3D rendered image of stained and embedded biological tissue. Structural information over volumes of tens of thousands of cubic micrometers is possible, revealing complex microanatomy with subcellular resolution. Methods are presented for tissue processing, for the enhancement of contrast with osmium tetroxide/potassium ferricyanide, for BSE imaging, for the preparation and platinum deposition over a selected site in the embedded tissue block, and for sequential data collection with ion beam milling; all this takes approximately 90 h. The imaging conditions, procedures for alternate milling and data acquisition and techniques for processing and partitioning the 3D data set are also described; these processes take approxiamtely 30 h. The protocol is illustrated by application to developing chick cornea, in which cells organize collagen fibril bundles into complex, multilamellar structures essential for transparency in the mature connective tissue matrix. The techniques described could have wide application in a range of fields, including pathology, developmental biology, microstructural anatomy and regenerative medicine. PMID- 21637204 TI - Monitoring protein expression in whole-cell extracts by targeted label- and standard-free LC-MS/MS. AB - Targeted quantification of proteins is a daily task in biological research but often relies on techniques such as western blotting that are only barely quantitative. Here we present a broadly applicable workflow for protein quantification from unpurified whole-cell extracts that can be completed in less than 3 d. Without prefractionation or affinity enrichment, a whole-cell extract is trypsin-digested in an acetonitrile-containing ammonium carbonate buffer and high-molecular-weight compounds are removed by filtration. A normalization strategy, which involves endogenous reference proteins, facilitates the determination of relative changes in protein expression without requiring isotope labeling or standard addition. On a triple-quadrupole mass spectrometer, we demonstrate standard-free quantification of yeast proteins present over five orders of magnitude and present at >=500 copies per cell. Liquid chromatography/multiple reaction monitoring (LC-MRM)-based proteomics is therefore a next-generation alternative to western blotting, as it allows simultaneous and reliable quantification of multiple endogenous proteins without the need for enrichment, isotope labeling or use of antibodies. PMID- 21637205 TI - Genome-scale analysis of replication timing: from bench to bioinformatics. AB - Replication timing profiles are cell type-specific and reflect genome organization changes during differentiation. In this protocol, we describe how to analyze genome-wide replication timing (RT) in mammalian cells. Asynchronously cycling cells are pulse labeled with the nucleotide analog 5-bromo-2-deoxyuridine (BrdU) and sorted into S-phase fractions on the basis of DNA content using flow cytometry. BrdU-labeled DNA from each fraction is immunoprecipitated, amplified, differentially labeled and co-hybridized to a whole-genome comparative genomic hybridization microarray, which is currently more cost effective than high throughput sequencing and equally capable of resolving features at the biologically relevant level of tens to hundreds of kilobases. We also present a guide to analyzing the resulting data sets based on methods we use routinely. Subjects include normalization, scaling and data quality measures, LOESS (local polynomial) smoothing of RT values, segmentation of data into domains and assignment of timing values to gene promoters. Finally, we cover clustering methods and means to relate changes in the replication program to gene expression and other genetic and epigenetic data sets. Some experience with R or similar programming languages is assumed. All together, the protocol takes ~3 weeks per batch of samples. PMID- 21637206 TI - In vivo solid-phase microextraction for monitoring intravenous concentrations of drugs and metabolites. AB - This protocol for in vivo solid-phase microextraction (SPME) can be used to monitor and quantify intravenous concentrations of drugs and metabolites without the need to withdraw a blood sample for analysis. The SPME probe is inserted directly into a peripheral vein of a living animal through a standard medical catheter, and extraction occurs typically over 2-5 min. After extraction, the analytes are removed from the sorbent and analyzed by, for example, liquid chromatography-tandem mass spectrometry. It has been validated in comparison with conventional blood analysis, and we describe here the in vitro experiments typically conducted during method development. The new-generation biocompatible SPME probes are designed specifically for extraction of semi-volatiles and nonvolatiles directly from aqueous samples and can be steam sterilized. Sorbents are coated on fine-gauge surgical steel wire (200-MUm diameter), which is more rugged and biocompatible than conventional fibers (100-MUm fused silica fiber). They incorporate a binding agent that resists fouling by the biological matrix and does not cause an immune response in the experimental animal. The sorbents used (coating thickness of ~50 MUm) are selected for their affinity for the types of small molecules of interest. The procedure is illustrated by the analysis of benzodiazepines with polypyrrole-coated wires inserted into peripheral blood vessels of beagles, although it can be adapted for use in smaller animals. The in vivo sampling can require as little as 1 min, in which case the entire procedure from sampling to instrumental analysis can take as little as 30 min. PMID- 21637207 TI - A time-of-drug addition approach to target identification of antiviral compounds. AB - Insight into the mode of action of newly discovered antiviral agents is now almost a prerequisite for clinical development. This protocol describes a method that provides information on the target of inhibitors of the human immunodeficiency virus (HIV); it can also be adapted to other viruses. The results from this experiment are available within 2 d. This time-based approach determines how long the addition of a compound can be postponed before losing its antiviral activity in cell culture. The target of an antiviral compound can be identified by comparing its relative position in the time scale to that of reference drugs. Therefore, it is more precise than, for example, in the case of HIV, a determination of pre- or postintegrational mode of action, and combines in one routine different assays for studying mechanisms of action. PMID- 21637209 TI - Mineralogical constraints on Precambrian pCO2. PMID- 21637210 TI - Faint young Sun paradox remains. PMID- 21637211 TI - Low pCO2 in the pore water, not in the Archean air. PMID- 21637212 TI - A big disease with a little name. PMID- 21637213 TI - Gold mine. PMID- 21637214 TI - Board the eurostar. PMID- 21637215 TI - China needs no foreign help to feed itself. PMID- 21637227 TI - A new eye on biodiversity. PMID- 21637228 TI - Scientists on trial over L'Aquila deaths. PMID- 21637229 TI - Japan quake rocks fusion project. PMID- 21637230 TI - Tevatron's legacy set to disappear. PMID- 21637231 TI - Furore erupts over Turkish test cipher. PMID- 21637232 TI - First sale for quantum computing. PMID- 21637233 TI - Will you take the 'arsenic-life' test? PMID- 21637234 TI - Physics meets cancer: The disruptor. PMID- 21637235 TI - Quantum computing: The power of discord. PMID- 21637236 TI - Children of the AIDS pandemic. PMID- 21637237 TI - Stigma impedes AIDS prevention. PMID- 21637242 TI - Population mapping of poor countries. PMID- 21637243 TI - Cameroon: listening to indigenous peoples. PMID- 21637244 TI - Respect limits of embryo patents. PMID- 21637246 TI - Willard Boyle (1924-2011). PMID- 21637245 TI - Cameroon: not ready for REDD+. PMID- 21637247 TI - Malaria: Mosquitoes bamboozled. PMID- 21637248 TI - Quantum information: Entanglement as elbow grease. PMID- 21637249 TI - Immunology: In command of commensals. PMID- 21637250 TI - Palaeoanthropology: In search of the australopithecines. PMID- 21637251 TI - Molecular evolution: Hidden diversity sparks adaptation. PMID- 21637252 TI - Climate change: Ancient Antarctic fjords. PMID- 21637254 TI - The thermodynamic meaning of negative entropy. AB - The heat generated by computations is not only an obstacle to circuit miniaturization but also a fundamental aspect of the relationship between information theory and thermodynamics. In principle, reversible operations may be performed at no energy cost; given that irreversible computations can always be decomposed into reversible operations followed by the erasure of data, the problem of calculating their energy cost is reduced to the study of erasure. Landauer's principle states that the erasure of data stored in a system has an inherent work cost and therefore dissipates heat. However, this consideration assumes that the information about the system to be erased is classical, and does not extend to the general case where an observer may have quantum information about the system to be erased, for instance by means of a quantum memory entangled with the system. Here we show that the standard formulation and implications of Landauer's principle are no longer valid in the presence of quantum information. Our main result is that the work cost of erasure is determined by the entropy of the system, conditioned on the quantum information an observer has about it. In other words, the more an observer knows about the system, the less it costs to erase it. This result gives a direct thermodynamic significance to conditional entropies, originally introduced in information theory. Furthermore, it provides new bounds on the heat generation of computations: because conditional entropies can become negative in the quantum case, an observer who is strongly correlated with a system may gain work while erasing it, thereby cooling the environment. PMID- 21637253 TI - Crystal structure of the FimD usher bound to its cognate FimC-FimH substrate. AB - Type 1 pili are the archetypal representative of a widespread class of adhesive multisubunit fibres in Gram-negative bacteria. During pilus assembly, subunits dock as chaperone-bound complexes to an usher, which catalyses their polymerization and mediates pilus translocation across the outer membrane. Here we report the crystal structure of the full-length FimD usher bound to the FimC FimH chaperone-adhesin complex and that of the unbound form of the FimD translocation domain. The FimD-FimC-FimH structure shows FimH inserted inside the FimD 24-stranded beta-barrel translocation channel. FimC-FimH is held in place through interactions with the two carboxy-terminal periplasmic domains of FimD, a binding mode confirmed in solution by electron paramagnetic resonance spectroscopy. To accommodate FimH, the usher plug domain is displaced from the barrel lumen to the periplasm, concomitant with a marked conformational change in the beta-barrel. The amino-terminal domain of FimD is observed in an ideal position to catalyse incorporation of a newly recruited chaperone-subunit complex. The FimD-FimC-FimH structure provides unique insights into the pilus subunit incorporation cycle, and captures the first view of a protein transporter in the act of secreting its cognate substrate. PMID- 21637255 TI - A dynamic early East Antarctic Ice Sheet suggested by ice-covered fjord landscapes. AB - The first Cenozoic ice sheets initiated in Antarctica from the Gamburtsev Subglacial Mountains and other highlands as a result of rapid global cooling ~34 million years ago. In the subsequent 20 million years, at a time of declining atmospheric carbon dioxide concentrations and an evolving Antarctic circumpolar current, sedimentary sequence interpretation and numerical modelling suggest that cyclical periods of ice-sheet expansion to the continental margin, followed by retreat to the subglacial highlands, occurred up to thirty times. These fluctuations were paced by orbital changes and were a major influence on global sea levels. Ice-sheet models show that the nature of such oscillations is critically dependent on the pattern and extent of Antarctic topographic lowlands. Here we show that the basal topography of the Aurora Subglacial Basin of East Antarctica, at present overlain by 2-4.5 km of ice, is characterized by a series of well-defined topographic channels within a mountain block landscape. The identification of this fjord landscape, based on new data from ice-penetrating radar, provides an improved understanding of the topography of the Aurora Subglacial Basin and its surroundings, and reveals a complex surface sculpted by a succession of ice-sheet configurations substantially different from today's. At different stages during its fluctuations, the edge of the East Antarctic Ice Sheet lay pinned along the margins of the Aurora Subglacial Basin, the upland boundaries of which are currently above sea level and the deepest parts of which are more than 1 km below sea level. Although the timing of the channel incision remains uncertain, our results suggest that the fjord landscape was carved by at least two iceflow regimes of different scales and directions, each of which would have over-deepened existing topographic depressions, reversing valley floor slopes. PMID- 21637256 TI - Strontium isotope evidence for landscape use by early hominins. AB - Ranging and residence patterns among early hominins have been indirectly inferred from morphology, stone-tool sourcing, referential models and phylogenetic models. However, the highly uncertain nature of such reconstructions limits our understanding of early hominin ecology, biology, social structure and evolution. We investigated landscape use in Australopithecus africanus and Paranthropus robustus from the Sterkfontein and Swartkrans cave sites in South Africa using strontium isotope analysis, a method that can help to identify the geological substrate on which an animal lived during tooth mineralization. Here we show that a higher proportion of small hominins than large hominins had non-local strontium isotope compositions. Given the relatively high levels of sexual dimorphism in early hominins, the smaller teeth are likely to represent female individuals, thus indicating that females were more likely than males to disperse from their natal groups. This is similar to the dispersal pattern found in chimpanzees, bonobos and many human groups, but dissimilar from that of most gorillas and other primates. The small proportion of demonstrably non-local large hominin individuals could indicate that male australopiths had relatively small home ranges, or that they preferred dolomitic landscapes. PMID- 21637257 TI - Nematoda from the terrestrial deep subsurface of South Africa. AB - Since its discovery over two decades ago, the deep subsurface biosphere has been considered to be the realm of single-cell organisms, extending over three kilometres into the Earth's crust and comprising a significant fraction of the global biosphere. The constraints of temperature, energy, dioxygen and space seemed to preclude the possibility of more-complex, multicellular organisms from surviving at these depths. Here we report species of the phylum Nematoda that have been detected in or recovered from 0.9-3.6-kilometre-deep fracture water in the deep mines of South Africa but have not been detected in the mining water. These subsurface nematodes, including a new species, Halicephalobus mephisto, tolerate high temperature, reproduce asexually and preferentially feed upon subsurface bacteria. Carbon-14 data indicate that the fracture water in which the nematodes reside is 3,000-12,000-year-old palaeometeoric water. Our data suggest that nematodes should be found in other deep hypoxic settings where temperature permits, and that they may control the microbial population density by grazing on fracture surface biofilm patches. Our results expand the known metazoan biosphere and demonstrate that deep ecosystems are more complex than previously accepted. The discovery of multicellular life in the deep subsurface of the Earth also has important implications for the search for subsurface life on other planets in our Solar System. PMID- 21637258 TI - Ultra-prolonged activation of CO2-sensing neurons disorients mosquitoes. AB - Carbon dioxide (CO(2)) present in exhaled air is the most important sensory cue for female blood-feeding mosquitoes, causing activation of long-distance host seeking flight, navigation towards the vertebrate host and, in the case of Aedes aegypti, increased sensitivity to skin odours. The CO(2) detection machinery is therefore an ideal target to disrupt host seeking. Here we use electrophysiological assays to identify a volatile odorant that causes an unusual, ultra-prolonged activation of CO(2)-detecting neurons in three major disease-transmitting mosquitoes: Anopheles gambiae, Culex quinquefasciatus and A. aegypti. Importantly, ultra-prolonged activation of these neurons severely compromises their ability subsequently to detect CO(2) for several minutes. We also identify odours that strongly inhibit CO(2)-sensitive neurons as candidates for use in disruption of host-seeking behaviour, as well as an odour that evokes CO(2)-like activity and thus has potential use as a lure in trapping devices. Analysis of responses to panels of structurally related odours across the three mosquitoes and Drosophila, which have related CO(2)-receptor proteins, reveals a pattern of inhibition that is often conserved. We use video tracking in wind tunnel experiments to demonstrate that the novel ultra-prolonged activators can completely disrupt CO(2)-mediated activation as well as source-finding behaviour in Aedes mosquitoes, even after the odour is no longer present. Lastly, semi field studies demonstrate that use of ultra-prolonged activators disrupts CO(2) mediated hut entry behaviour of Culex mosquitoes. The three classes of CO(2) response-modifying odours offer powerful instruments for developing new generations of insect repellents and lures, which even in small quantities can interfere with the ability of mosquitoes to seek humans. PMID- 21637259 TI - Cryptic genetic variation promotes rapid evolutionary adaptation in an RNA enzyme. AB - Cryptic variation is caused by the robustness of phenotypes to mutations. Cryptic variation has no effect on phenotypes in a given genetic or environmental background, but it can have effects after mutations or environmental change. Because evolutionary adaptation by natural selection requires phenotypic variation, phenotypically revealed cryptic genetic variation may facilitate evolutionary adaptation. This is possible if the cryptic variation happens to be pre-adapted, or "exapted", to a new environment, and is thus advantageous once revealed. However, this facilitating role for cryptic variation has not been proven, partly because most pertinent work focuses on complex phenotypes of whole organisms whose genetic basis is incompletely understood. Here we show that populations of RNA enzymes with accumulated cryptic variation adapt more rapidly to a new substrate than a population without cryptic variation. A detailed analysis of our evolving RNA populations in genotype space shows that cryptic variation allows a population to explore new genotypes that become adaptive only in a new environment. Our observations show that cryptic variation contains new genotypes pre-adapted to a changed environment. Our results highlight the positive role that robustness and epistasis can have in adaptive evolution. PMID- 21637262 TI - ISCoS and WHO, an ever-strengthening cooperation for the benefit of individuals with spinal cord injuries worldwide. PMID- 21637264 TI - The FDA's generic-drug approval process: similarities to and differences from brand-name drugs. PMID- 21637265 TI - Algorithm for the management of endoscopic perforations: a quality improvement project. AB - Perforations are an uncommon but serious complication of endoscopy. Although they are well recognized, no universally accepted strategy for their management exists. The need for management algorithms in situations that call for multiple interventions in a short time, with coordinated effort encompassing multiple providers from different specialties, has long been recognized, but no such clinical care pathway has been developed for the management of endoscopic perforations. Since perforations are uncommon, a predetermined plan of action can streamline patient management. Furthermore, such a plan demonstrates preparedness on the part of the gastroenterologist. We developed an endoscopic perforation management strategy based on the best available scientific evidence and our specific resources. We report our experience in the hope that it may form a useful framework for gastroenterologists attempting to do the same at their own institution. PMID- 21637266 TI - Denial: what is it, how do we recognize it, and what should we do about it? PMID- 21637269 TI - Editorial: risk scoring for colon cancer screening: validated, but still not ready for prime time. AB - Risk stratification for colorectal cancer screening would allow us to use less expensive screening tests, such as sigmoidoscopy with or without fecal blood testing, on lower risk individuals, and reserve colonoscopy for those at higher risk. In this issue, Levitzky et al. validates a risk score that was previously developed by Imperiale et al., finding similar results among three ethnic groups. Risk scoring would detect 82-87% of proximal advanced neoplasia while decreasing colonoscopy use by 33-46%. However, before risk scoring is ready for widespread use, sigmoidoscopy access and performance issues need to be addressed, and we must be comfortable with missing some proximal neoplasms. PMID- 21637272 TI - The role of Acid in functional dyspepsia. PMID- 21637274 TI - Classification of the severity of acute pancreatitis. PMID- 21637276 TI - Small intestinal bacterial overgrowth and Helicobacter pylori: can they be cause of thrombocytopenia in patients with chronic liver disease? PMID- 21637277 TI - High prevalence of significant endoscopic findings in patients with uninvestigated typical reflux symptoms. PMID- 21637280 TI - Antibiotics and oral contraceptive efficacy in inflammatory bowel disease. PMID- 21637283 TI - Stem cell mobilization in patients with newly diagnosed multiple myeloma after lenalidomide induction therapy. AB - Lenalidomide has raised concerns regarding its potential impact on the ability to collect stem cells for autologous stem cell transplantation, especially after prolonged exposure. The use of cyclophosphamide plus granulocyte colony stimulating factor (G-CSF) to mobilize peripheral blood stem cells may overcome this concern. In newly diagnosed multiple myeloma (MM) patients, we investigated the influence of lenalidomide on stem cell collection. In a prospective study, 346 patients received four cycles of lenalidomide-dexamethasone (Rd). Stem cells were mobilized with cyclophosphamide and G-CSF. Patients failing to collect a minimum of 4 * 10(6) CD34(+)/kg cells received a second mobilization course. After mobilization, a median yield of 8.7 * 10(6) CD34(+)/kg was obtained from patients receiving Rd induction. After first mobilization, inadequate yield was observed in 21% of patients, whereas only 9% of patients failed to collect the target yield after the second mobilization attempt. In conclusion, we confirm that a short induction with lenalidomide allowed sufficient stem cells collection to perform autologous transplantation in 91% of newly diagnosed patients. PMID- 21637284 TI - Identification of a role for the nuclear receptor EAR-2 in the maintenance of clonogenic status within the leukemia cell hierarchy. AB - Identification of genes that regulate clonogenicity of acute myelogenous leukemia (AML) cells is hindered by the difficulty of isolating pure populations of cells with defined proliferative abilities. By analyzing the growth of clonal siblings in low passage cultures of the cell line OCI/AML4 we resolved this heterogeneous population into strata of distinct clonogenic potential, permitting analysis of the transcriptional signature of single cells with defined proliferative abilities. By microarray analysis we showed that the expression of the orphan nuclear receptor EAR-2 (NR2F6) is greater in leukemia cells with extensive proliferative capacity than in those that have lost proliferative ability. EAR-2 is expressed highly in long-term hematopoietic stem cells, relative to short-term hematopoietic stem and progenitor cells, and is downregulated in AML cells after induction of differentiation. Exogenous expression of EAR-2 increased the growth of U937 cells and prevented the proliferative arrest associated with terminal differentiation, and blocked differentiation of U937 and 32Dcl3 cells. Conversely, silencing of EAR-2 by short-hairpin RNA initiated terminal differentiation of these cell lines. These data identify EAR-2 as an important factor in the regulation of clonogenicity and differentiation, and establish that analysis of clonal siblings allows the elucidation of differences in gene expression within the AML hierarchy. PMID- 21637285 TI - The emerging role of hypoxia, HIF-1 and HIF-2 in multiple myeloma. AB - Hypoxia is an imbalance between oxygen supply and demand, which deprives cells or tissues of sufficient oxygen. It is well-established that hypoxia triggers adaptive responses, which contribute to short- and long-term pathologies such as inflammation, cardiovascular disease and cancer. Induced by both microenvironmental hypoxia and genetic mutations, the elevated expression of the hypoxia-inducible transcription factor-1 (HIF-1) and HIF-2 is a key feature of many human cancers and has been shown to promote cellular processes, which facilitate tumor progression. In this review, we discuss the emerging role of hypoxia and the HIFs in the pathogenesis of multiple myeloma (MM), an incurable hematological malignancy of BM PCs, which reside within the hypoxic BM microenvironment. The need for current and future therapeutic interventions to target HIF-1 and HIF-2 in myeloma will also be discussed. PMID- 21637286 TI - Mutation analysis of TET2, IDH1, IDH2 and ASXL1 in chronic myeloid leukemia. PMID- 21637287 TI - 18th Euroconference on Apoptosis in Ghent, Belgium, 1-4 September 2010. PMID- 21637288 TI - The role of vacuole in plant cell death. AB - Almost all plant cells have large vacuoles that contain both hydrolytic enzymes and a variety of defense proteins. Plants use vacuoles and vacuolar contents for programmed cell death (PCD) in two different ways: for a destructive way and for a non-destructive way. Destruction is caused by vacuolar membrane collapse, followed by the release of vacuolar hydrolytic enzymes into the cytosol, resulting in rapid and direct cell death. The destructive way is effective in the digestion of viruses proliferating in the cytosol, in susceptible cell death induced by fungal toxins, and in developmental cell death to generate integuments (seed coats) and tracheary elements. On the other hand, the non-destructive way involves fusion of the vacuolar and the plasma membrane, which allows vacuolar defense proteins to be discharged into the extracellular space where the bacteria proliferate. Membrane fusion, which is normally suppressed, was triggered in a proteasome-dependent manner. Intriguingly, both ways use enzymes with caspase like activity; the membrane-fusion system uses proteasome subunit PBA1 with caspase-3-like activity, and the vacuolar-collapse system uses vacuolar processing enzyme (VPE) with caspase-1-like activity. This review summarizes two different ways of vacuole-mediated PCD and discusses how plants use them to attack pathogens that invade unexpectedly. PMID- 21637289 TI - DeltaNp63alpha regulates keratinocyte proliferation by controlling PTEN expression and localization. AB - DeltaNp63alpha, implicated as an oncogene, is upregulated by activated Akt, part of a well-known cell survival pathway. Inhibition of Akt activation by phosphatase and tensin homolog deleted on chromosome 10 (PTEN) and the presence of putative p63-binding sites in the pten promoter led us to investigate whether DeltaNp63alpha regulates PTEN expression. Knockdown of DeltaNp63alpha led to increases in PTEN levels and loss of activated Akt, while overexpression of DeltaNp63alpha decreased PTEN levels and elevated active Akt. The repression of PTEN by DeltaNp63alpha occurs independently of p53 status, as loss of DeltaNp63alpha increases PTEN expression in cell lines with and without functional p53. In addition, decreased levels of DeltaNp63alpha resulted in an increase in nuclear PTEN. Conversely, in vivo nuclear PTEN was absent in the proliferative basal layer of the epidermis where DeltaNp63alpha expression is highest. Additionally, we show that in keratinocytes a balance between DeltaNp63alpha and PTEN regulates Akt activation and maintains normal proliferation rates. This balance is disrupted in non-melanoma skin cancers through increased DeltaNp63alpha levels, and could enhance proliferation and subsequent neoplastic development. Our studies show that DeltaNp63alpha negatively regulates PTEN, thereby providing a feedback loop between PTEN, Akt and DeltaNp63alpha, which has an integral role in skin cancer development. PMID- 21637290 TI - SAHA shows preferential cytotoxicity in mutant p53 cancer cells by destabilizing mutant p53 through inhibition of the HDAC6-Hsp90 chaperone axis. AB - Mutant p53 (mutp53) cancers are surprisingly dependent on their hyperstable mutp53 protein for survival, identifying mutp53 as a potentially significant clinical target. However, exploration of effective small molecule therapies targeting mutp53 has barely begun. Mutp53 hyperstabilization, a hallmark of p53 mutation, is cancer cell-specific and due to massive upregulation of the HSP90 chaperone machinery during malignant transformation. We recently showed that stable complex formation between HSP90 and its mutp53 client inhibits E3 ligases MDM2 and CHIP, causing mutp53 stabilization. Histone deacetylase (HDAC) inhibitors (HDACi) are a new class of promising anti-cancer drugs, hyperacetylating histone and non-histone targets. Currently, suberoylanilide hydroxamic acid (SAHA) is the only FDA-approved HDACi. We show that SAHA exhibits preferential cytotoxicity for mutant, rather than wild-type and null p53 human cancer cells. Loss/gain-of-function experiments revealed that although able to exert multiple cellular effects, SAHA's cytotoxicity is caused to a significant degree by its ability to strongly destabilize mutp53 at the level of protein degradation. The underlying mechanism is SAHA's inhibition of HDAC6, an essential positive regulator of HSP90. This releases mutp53 and enables its MDM2- and CHIP mediated degradation. SAHA also strongly chemosensitizes mutp53 cancer cells for chemotherapy due to its ability to degrade mutp53. This identifies a novel action of SAHA with the prospect of SAHA becoming a centerpiece in mutp53-specific anticancer strategies. PMID- 21637291 TI - Mitochondrially localized PKA reverses mitochondrial pathology and dysfunction in a cellular model of Parkinson's disease. AB - Mutations in PTEN-induced kinase 1 (PINK1) are associated with a familial syndrome related to Parkinson's disease (PD). We previously reported that stable neuroblastoma SH-SY5Y cell lines with reduced expression of endogenous PINK1 exhibit mitochondrial fragmentation, increased mitochondria-derived superoxide, induction of compensatory macroautophagy/mitophagy and a low level of ongoing cell death. In this study, we investigated the ability of protein kinase A (PKA) to confer protection in this model, focusing on its subcellular targeting. Either: (1) treatment with pharmacological PKA activators; (2) transient expression of a constitutively active form of mitochondria-targeted PKA; or (3) transient expression of wild-type A kinase anchoring protein 1 (AKAP1), a scaffold that targets endogenous PKA to mitochondria, reversed each of the phenotypes attributed to loss of PINK1 in SH-SY5Y cells, and rescued parameters of mitochondrial respiratory dysfunction. Mitochondrial and lysosomal changes in primary cortical neurons derived from PINK1 knockout mice or subjected to PINK1 RNAi were also reversed by the activation of PKA. PKA phosphorylates the rat dynamin-related protein 1 isoform 1 (Drp1) at serine 656 (homologous to human serine 637), inhibiting its pro-fission function. Mimicking phosphorylation of Drp1 recapitulated many of the protective effects of AKAP1/PKA. These data indicate that redirecting endogenous PKA to mitochondria can compensate for deficiencies in PINK1 function, highlighting the importance of compartmentalized signaling networks in mitochondrial quality control. PMID- 21637292 TI - Peculiarities of cell death mechanisms in neutrophils. AB - Analyses of neutrophil death mechanisms have revealed many similarities with other cell types; however, a few important molecular features make these cells unique executors of cell death mechanisms. For instance, in order to fight invading pathogens, neutrophils possess a potent machinery to produce reactive oxygen species (ROS), the phagocyte nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. Evidence is emerging that these ROS are crucial in the execution of most neutrophil cell death mechanisms. Likewise, neutrophils exhibit many diverse granules that are packed with cytotoxic mediators. Of those, cathepsins were recently shown to activate pro-apoptotic B-cell lymphoma-2 (Bcl-2) family members and caspases, thus acting on apoptosis regulators. Moreover, neutrophils have few mitochondria, which hardly participate in ATP synthesis, as neutrophils gain energy from glycolysis. In spite of relatively low levels of cytochrome c in these cells, the mitochondrial death pathway is functional. In addition to these pecularities defining neutrophil death pathways, neutrophils are terminally differentiated cells, hence they do not divide but undergo apoptosis shortly after maturation. The initial trigger of this spontaneous apoptosis remains to be determined, but may result from low transcription and translation activities in mature neutrophils. Due to the unique biological characteristics of neutrophils, pharmacological intervention of inflammation has revealed unexpected and sometimes disappointing results when neutrophils were among the prime target cells during therapy. In this study, we review the current and emerging models of neutrophil cell death mechanisms with a focus on neutrophil peculiarities. PMID- 21637293 TI - Absolute requirement for STAT3 function in small-intestine crypt stem cell survival. AB - The transcription factor signal transducer and activator of transcription 3 (STAT3) is frequently activated in human cancers. Interestingly, STAT3 also maintains the pluripotency and self-renewal of murine embryonic stem cells, and several tissue stem cell types. To investigate whether STAT3 also maintains the small-intestine crypt stem cell, we conditionally inactivated a Floxed Stat3 allele (Stat3(fl)) in murine small-intestine crypt stem cells. Following Cre recombinase expression, apoptosis increased in Stat3(fl/-) experimental crypts relative to Stat3(wt/-) controls before declining. Control Stat3(wt/-) mice carrying a Flox-STOP LacZ reporter transgene stably expressed LacZ after Cre induction. In contrast, Stat3(fl/-) intestine LacZ expression initially increased modestly, before declining to background levels. Quantitative PCRs revealed a similar transient in recombined Stat3(fl) allele levels. Long-term bromodeoxyuridine labelling directly demonstrated that functional STAT3 is required for +4 to +6 region label-retaining small-intestine stem cell survival. Rapid clearance of recombined Stat3(fl/-) cells involves apoptosis potentially induced by elevated c-Myc in non-recombined cells and involves elevated p53 expression and caspase 3 activation. Intriguingly, Stat3(fl/-) intestine recombination triggered dramatically upregulated polycomb transcriptional repressor Bmi1 - potentially accelerating recombined crypt repopulation. In summary, STAT3 activity is absolutely required for small-intestine crypt stem cell survival at both the +4 to +6 label-retaining and crypt base columnar cell locations. PMID- 21637294 TI - Ret finger protein inhibits muscle differentiation by modulating serum response factor and enhancer of polycomb1. AB - Skeletal myogenesis is precisely regulated by multiple transcription factors. Previously, we demonstrated that enhancer of polycomb 1 (Epc1) induces skeletal muscle differentiation by potentiating serum response factor (SRF)-dependent muscle gene activation. Here, we report that an interacting partner of Epc1, ret finger protein (RFP), blocks skeletal muscle differentiation. Our findings show that RFP was highly expressed in skeletal muscles and was downregulated during myoblast differentiation. Forced expression of RFP delayed myoblast differentiation, whereas knockdown enhanced it. Epc1-induced enhancements of SRF dependent multinucleation, transactivation of the skeletal alpha-actin promoter, binding of SRF to the serum response element, and muscle-specific gene induction were blocked by RFP. RFP interfered with the physical interaction between Epc1 and SRF. Muscles from rfp knockout mice (Rfp(-/-)) mice were bigger than those from wild-type mice, and the expression of SRF-dependent muscle-specific genes was upregulated. Myotube formation and myoblast differentiation were enhanced in Rfp(-/-) mice. Taken together, our findings highlight RFP as a novel regulator of muscle differentiation that acts by modulating the expression of SRF-dependent skeletal muscle-specific genes. PMID- 21637295 TI - RNA steady-state defects in myotonic dystrophy are linked to nuclear exclusion of SHARP. AB - We describe a new mechanism by which CTG tract expansion affects myotonic dystrophy (DM1). Changes to the levels of a panel of RNAs involved in muscle development and function that are downregulated in DM1 are due to aberrant localization of the transcription factor SHARP (SMART/HDAC1-associated repressor protein). Mislocalization of SHARP in DM1 is consistent with increased CRM1 mediated export of SHARP to the cytoplasm. A direct link between CTG repeat expression and SHARP mislocalization is demonstrated as expression of expanded CTG repeats in normal cells recapitulates cytoplasmic SHARP localization. These results demonstrate a role for the inactivation of SHARP transcription in DM1 biology. PMID- 21637296 TI - Single-molecule approach to immunoprecipitated protein complexes: insights into miRNA uridylation. AB - Single-molecule techniques have been used for only a subset of biological problems because of difficulties in studying proteins that require cofactors or post-translational modifications. Here, we present a new method integrating single-molecule fluorescence microscopy and immunopurification to study protein complexes. We used this method to investigate Lin28-mediated microRNA uridylation by TUT4 (terminal uridylyl transferase 4, polyU polymerase), which regulates let 7 microRNA biogenesis. Our real-time analysis of the uridylation by the TUT4 immunoprecipitates suggests that Lin28 functions as a processivity factor of TUT4. Our new technique, SIMPlex (single-molecule approach to immunoprecipitated protein complexes), provides a universal tool to analyse complex proteins at the single-molecule level. PMID- 21637297 TI - p53 downregulates Down syndrome-associated DYRK1A through miR-1246. AB - Several microRNAs mediate the functions of p53 family members. Here we characterize miR-1246 as a new target of this family. In response to DNA damage, p53 induces the expression of miR-1246 which, in turn, reduces the level of DYRK1A, a Down syndrome-associated protein kinase. Knockdown of p53 has the opposite effect. Overexpression of miR-1246 reduces DYRK1A levels and leads to the nuclear retention of NFATc1, a protein substrate of DYRK1A, and the induction of apoptosis, whereas a miR-1246-specific inhibitor prevented the nuclear import of NFATc1. Together, these results indicate that p53 inhibits DYRK1A expression through the induction of miR-1246. PMID- 21637298 TI - ATM-mediated phosphorylation of polynucleotide kinase/phosphatase is required for effective DNA double-strand break repair. AB - The cellular response to double-strand breaks (DSBs) in DNA is a complex signalling network, mobilized by the nuclear protein kinase ataxia-telangiectasia mutated (ATM), which phosphorylates many factors in the various branches of this network. A main question is how ATM regulates DSB repair. Here, we identify the DNA repair enzyme polynucleotide kinase/phosphatase (PNKP) as an ATM target. PNKP phosphorylates 5'-OH and dephosphorylates 3'-phosphate DNA ends that are formed at DSB termini caused by DNA-damaging agents, thereby regenerating legitimate ends for further processing. We establish that the ATM phosphorylation targets on human PNKP-Ser 114 and Ser 126-are crucial for cellular survival following DSB induction and for effective DSB repair, being essential for damage-induced enhancement of the activity of PNKP and its proper accumulation at the sites of DNA damage. These findings show a direct functional link between ATM and the DSB repair machinery. PMID- 21637299 TI - A genetic screen identifies BRCA2 and PALB2 as key regulators of G2 checkpoint maintenance. AB - To identify key connections between DNA-damage repair and checkpoint pathways, we performed RNA interference screens for regulators of the ionizing radiation induced G2 checkpoint, and we identified the breast cancer gene BRCA2. The checkpoint was also abrogated following depletion of PALB2, an interaction partner of BRCA2. BRCA2 and PALB2 depletion led to premature checkpoint abrogation and earlier activation of the AURORA A-PLK1 checkpoint-recovery pathway. These results indicate that the breast cancer tumour suppressors and homologous recombination repair proteins BRCA2 and PALB2 are main regulators of G2 checkpoint maintenance following DNA-damage. PMID- 21637300 TI - Microbiological cure times in acanthamoeba keratitis. AB - AIMS: The purpose of this study was to estimate the duration of treatment necessary for sequential acanthamoeba laboratory tests from corneal scrapings to become negative, and to assess predictors that affect this duration period. METHODS: We included all patients with at least one positive acanthamoeba culture or Giemsa stain at the F.I. Proctor Foundation Microbiology Laboratory from 1996 to 2009. A parametric survival analysis was performed among patients with repeat cultures to assess significant predictors for extended clearance time. Simulations were performed to estimate clearance time in the entire patient population, assuming imperfect sensitivity. RESULTS: Thirty-seven patients with laboratory evidence of acanthamoeba had testing at 69 time points. The median clearance time among eyes with repeat cultures was 42.5 days (interquartile range (IQR) 22.0-82.0 days; unadjusted parametric model). Initial visual acuity was the only predictor significantly associated with clearance time in univariate analyses (P<0.0001). Using initial visual acuity as a predictor for clearance time among the entire patient population, the estimated clearance time decreased to 38.7 days (95% confidence interval (CI) 27.9-53.5 days). When the imperfect sensitivity of the culture technique was also taken into account, the estimated clearance time was 44.1 days (95% CI 31.9-61.0 days). CONCLUSION: The duration of infection with acanthamoeba keratitis undergoing treatment has not been well characterized. In this report we estimate a median clearance time of approximately 6 weeks, with an IQR of 22-82 days. PMID- 21637301 TI - Safe eye surgery: non-technical aspects. AB - The traditional training of surgeons focused exclusively on developing knowledge, clinical expertise, and technical (surgical) skills. However, analyses of the reasons for adverse events in surgery have revealed that many underlying causes originate from behavioural or non-technical aspects of performance (eg, poor communication among members of the surgical team) rather than from a lack of surgical (ie, technical) skills. Therefore, technical skills appear to be necessary but not sufficient to ensure patient safety. Paying attention to non technical skills, such as team working, leadership, situation awareness, decision making, and communication, will increase the likelihood of maintaining high levels of error-free performance. Identification and training of non-technical skills has been developed for high-risk careers, such as civil aviation and nuclear power. Only recently, training in non-technical skills has been adopted by the surgical world and anaesthetists. Non-technical skills need to be tailored to the environment where they are used, and eye surgery has some substantial differences compared with other surgical areas, for example, high volume of surgery, use of local anaesthetics, and very sophisticated equipment. This review highlights the need for identification of the non-technical skills relevant to eye surgeons and promotion of their use in the training of eye surgeons. PMID- 21637302 TI - Comment on a new ocular trauma score in pediatric penetrating eye injuries. PMID- 21637304 TI - Trans-scleral dye injection during vitreous surgery to identify clinically undetectable retinal breaks causing retinal detachment. AB - BACKGROUND/AIMS: Finding all retinal breaks is a critical step in rhegmatogenous retinal detachment (RRD) surgery in order to prevent persistent/recurrent retinal detachment (RD). We describe a technique of trans-scleral dye injection into the subretinal fluid under the detached retina in the context of recurrent/persistent RD in vitrectomized eyes, in order to determine the location of clinically unidentified (occult) retinal breaks causing RD. METHODS: Retrospective consecutive single-surgeon case-series analysis of patients presenting with a repeat RRD after having been treated with pars plana vitrectomy (PPV) as the method of primary RRD repair. Trans-scleral injection of subretinal vision blue (TSVB) was used to help identify retinal breaks during repeat vitrectomy. OUTCOME MEASURES: successful detection of a break; location of breaks; persistent retinal attachment; final visual acuity (VA); complications. RESULTS: There were 395 cases of RRD during the 3-year period reviewed. TSVB was used for eight instances in seven eyes. All eight instances were repeat RRD. TSVB facilitated occult break detection in 7/8 instances of use. Breaks were at or adjacent to the previous cryo site in three instances. Persistent retinal attachment was achieved in 5/7 cases. Final VA increased in 5/7 cases. There was no evidence of complications as a result of TSVB injection. CONCLUSIONS: TSVB coupled with indentation to vent a plume of dye through an occult break during vitreous surgery is a relatively simple technique that may facilitate the identification of occult retinal breaks and help achieve anatomical success and functional success. PMID- 21637303 TI - Non-steroidal drug-induced glaucoma. AB - Numerous systemically used drugs are involved in drug-induced glaucoma. Most reported cases of non-steroidal drug-induced glaucoma are closed-angle glaucoma (CAG). Indeed, many routinely used drugs that have sympathomimetic or parasympatholytic properties can cause pupillary block CAG in individuals with narrow iridocorneal angle. The resulting acute glaucoma occurs much more commonly unilaterally and only rarely bilaterally. CAG secondary to sulfa drugs is a bilateral non-pupillary block type and is due to forward movement of iris-lens diaphragm, which occurs in individuals with narrow or open iridocorneal angle. A few agents, including antineoplastics, may induce open-angle glaucoma. In conclusion, the majority of cases with glaucoma secondary to non-steroidal medications are of the pupillary block closed-angle type and preventable if the at-risk patients are recognized and treated prophylactically. PMID- 21637306 TI - A rare case of endogenous Streptococcus group C endophthalmitis associated with cellulitis. PMID- 21637307 TI - Radiation enhances long-term metastasis potential of residual hepatocellular carcinoma in nude mice through TMPRSS4-induced epithelial-mesenchymal transition. AB - Recurrence and metastasis are frequently observed after radiotherapy for hepatocellular carcinoma (HCC), although upregulation of matrix metalloproteinases (MMPs) and vascular endothelial growth factor (VEGF) induced by radiation has been claimed to be involved, the mechanism is not clarified yet. In the present study, by using MHCC97L, a human HCC cell line with metastatic potential, and its xenograft in nude mice, we found that radiation induced a 48- to 72-h temporary increase in the expression of MMP-2 and VEGF both in vitro and in vivo, but only the in vitro invasiveness of MHCC97L cells was enhanced, while the in vivo metastatic potential of tumors was suppressed. Whereas, 30 days after radiation, when the expression of MMP-2 and VEGF decreased to unirradiated control levels, the in vivo dissemination and metastatic potential of residual tumors have just begun to increase with overexpression of TMPRSS4, which induced loss of E-cadherin through induction of Smad-Interacting Protein 1 (SIP1), an E cadherin transcriptional repressor, and led to epithelial-mesenchymal transition (EMT). This process was blocked by treatment of siRNA-TMPRSS4. In conclusion, our study revealed novel findings regarding the biphasic effect of radiation on the metastatic potential of residual HCC. Overexpression of TMPRSS4 has a critical role in radiation-induced long-term dissemination and metastasis of residual HCC by facilitating EMT. These findings may provide new clues to suppress the radiation-induced dissemination and metastasis, thereby improve the prognosis of HCC patients. PMID- 21637308 TI - Experimental arthritis: Therapeutic promise of dual blockade of IL-17 and TNF in inflammatory arthritis. PMID- 21637309 TI - Connective tissue diseases: Notch signaling: an important player in SSc fibrosis. PMID- 21637310 TI - Therapy: Rituximab for refractory SLE--patients reach lasting remission with short-term regimen. PMID- 21637311 TI - Therapy: Atacicept lacks clinical efficacy in RA. PMID- 21637312 TI - Rheumatoid arthritis: Statins have immunomodulatory and clinical effects in RA. PMID- 21637313 TI - Therapy: Sodium oxybate for fibromyalgia. PMID- 21637315 TI - Vasculitis syndromes: T cells turned killer by IL-15 attack the endothelium in GPA. PMID- 21637316 TI - Clinical trials: Insufficient data on glucocorticoid use in RA trials. PMID- 21637317 TI - Susceptibility of patients with rheumatic diseases to B-cell non-Hodgkin lymphoma. AB - Autoimmune rheumatic diseases (ARD), such as systemic lupus erythematosus (SLE), rheumatoid arthritis (RA) and Sjogren's syndrome (SS), have consistently been associated with the development of B-cell non-Hodgkin lymphoma (BCNHL). In this Review, we focus on reports published since 2006 and summarize the data regarding the BCNHL subtypes and clinical findings associated with this increased risk. Patients with these ARD, particularly those with detectable autoantibodies and systemic involvement, are at increased risk of developing BCNHL, especially diffuse large B-cell lymphoma and marginal zone lymphoma. SS shows the strongest association with BCNHL. Male sex, advanced age, prolonged disease course and increased disease severity, but not family history of autoimmune conditions, seem to be associated with an increased risk of non-Hodgkin lymphoma. Chronic immune stimulation, genetic and environmental factors and some immunosuppressive drugs might be involved in lymphomagenesis in these patients. The reason why some ARD are associated with BCNHL and other autoimmune diseases are not remains unclear. These associations are important as they provide information about the mechanisms of lymphomagenesis, and might help identify new therapeutic targets. PMID- 21637318 TI - Scaling up services for mental and neurological disorders in low-resource settings. AB - Mental and neurological disorders (MNDs) account for a large, and growing, burden of disease in low- and middle-income countries. Most people do not have access to even basic health care for these disorders. Recent evidence shows that task shifting to non-specialist community health workers is a feasible and effective strategy for delivery of efficacious treatments for specific MND in low-resource settings. New global initiatives, such as the WHO's mental health Gap Action Program, are utilizing this evidence to devise packages of care for specific MNDs. This paper describes a plan that seeks to integrate the evidence on the treatment of specific MNDs, based on a task-shifting paradigm, for scaling up services for MNDs at the level of a defined population. The plan was developed by a state government in India in collaboration with technical partners, as a model District Mental Health Program for India's National Mental Health Program. PMID- 21637319 TI - An overview of ethnography in healthcare and medical education research. AB - Research in healthcare settings and medical education has relied heavily on quantitative methods. However, there are research questions within these academic domains that may be more adequately addressed by qualitative inquiry. While there are many qualitative approaches, ethnography is one method that allows the researcher to take advantage of relative immersion in order to obtain thick description. The purpose of this article is to introduce ethnography, to describe how ethnographic methods may be utilized, to provide an overview of ethnography's use in healthcare and medical education, and to summarize some key limitations with the method. PMID- 21637320 TI - The RUB Cage: Respiration-Ultrasonic Vocalizations-Behavior Acquisition Setup for Assessing Emotional Memory in Rats. AB - In animals, emotional memory is classically assessed through pavlovian fear conditioning in which a neutral novel stimulus (conditioned stimulus) is paired with an aversive unconditioned stimulus. After conditioning, the conditioned stimulus elicits a fear response characterized by a wide range of behavioral and physiological responses. Despite the existence of this large repertoire of responses, freezing behavior is often the sole parameter used for quantifying fear response, thus limiting emotional memory appraisal to this unique index. Interestingly, respiratory changes and ultrasonic vocalizations (USV) can occur during fear response, yet very few studies investigated the link between these different parameters and freezing. The aim of the present study was to design an experimental setup allowing the simultaneous recording of respiration, USV, and behavior (RUB cage), and the offline synchronization of the collected data for fine-grain second by second analysis. The setup consisted of a customized plethysmograph for respiration monitoring, equipped with a microphone capturing USV, and with four video cameras for behavior recording. In addition, the bottom of the plethysmograph was equipped with a shock-floor allowing foot-shock delivery, and the top received tubing for odor presentations. Using this experimental setup we first described the characteristics of respiration and USV in different behaviors and emotional states. Then we monitored these parameters during contextual fear conditioning and showed that they bring complementary information about the animal's anxiety state and the strength of aversive memory. The present setup may be valuable in providing a clearer appraisal of the physiological and behavioral changes that occur during acquisition as well as retrieval of emotional memory. PMID- 21637321 TI - Neural systems underlying aversive conditioning in humans with primary and secondary reinforcers. AB - Money is a secondary reinforcer commonly used across a range of disciplines in experimental paradigms investigating reward learning and decision-making. The effectiveness of monetary reinforcers during aversive learning and associated neural basis, however, remains a topic of debate. Specifically, it is unclear if the initial acquisition of aversive representations of monetary losses depends on similar neural systems as more traditional aversive conditioning that involves primary reinforcers. This study contrasts the efficacy of a biologically defined primary reinforcer (shock) and a socially defined secondary reinforcer (money) during aversive learning and its associated neural circuitry. During a two-part experiment, participants first played a gambling game where wins and losses were based on performance to gain an experimental bank. Participants were then exposed to two separate aversive conditioning sessions. In one session, a primary reinforcer (mild shock) served as an unconditioned stimulus (US) and was paired with one of two colored squares, the conditioned stimuli (CS+ and CS-, respectively). In another session, a secondary reinforcer (loss of money) served as the US and was paired with one of two different CS. Skin conductance responses were greater for CS+ compared to CS- trials irrespective of type of reinforcer. Neuroimaging results revealed that the striatum, a region typically linked with reward-related processing, was found to be involved in the acquisition of aversive conditioned response irrespective of reinforcer type. In contrast, the amygdala was involved during aversive conditioning with primary reinforcers, as suggested by both an exploratory fMRI analysis and a follow-up case study with a patient with bilateral amygdala damage. Taken together, these results suggest that learning about potential monetary losses may depend on reinforcement learning related systems, rather than on typical structures involved in more biologically based fears. PMID- 21637322 TI - Reliability of quadruplicated serological parameters in the korean genome and epidemiology study. AB - OBJECTIVES: The aim of this study was to evaluate whether clinical test values from different laboratories in the Korean Genome and Epidemiology Study (KoGES) can be integrated through a statistical adjustment algorithm with appropriate intra- and inter-laboratory reliability. METHODS: External quality control data were obtained from the Korean Society for Laboratory Medicine and quadruplicated standardized serological samples (N=3,200) were manufactured in order to check the intra- and inter-laboratory reliability for aspartic acid transaminase (AST), alanine transaminase (ALT), gamma-glutamyl transpeptidase (gamma-GTP), blood urea nitrogen (BUN), creatinine, uric acid (UA), fasting blood sugar (FBS), cholesterol, and triglyceride (TG). As an index of inter- and intra-rater reliability, Pearson's correlation coefficient, intraclass correlation coefficients and kappa statistics were estimated. In addition, to detect the potential for data integration, we constructed statistical compensation models using linear regression analysis with residual analysis, and presented the R square values. RESULTS: All correlation coefficient values indicated good intra- and inter-laboratory reliability, which ranged from 0.842 to 1.000. Kappa coefficients were greater than 0.75 (0.75-1.00). All of the regression models based on the trial results had strong R-square values and zero sums of residuals. These results were consistent in the regression models using external quality control data. CONCLUSION: The two laboratories in the KoGES have good intra- and inter-laboratory reliability for ten chemical test values, and data can be integrated through algorithmic statistical adjustment using regression equations. PMID- 21637323 TI - Basic helix-loop-helix transcription factor TCF21 is a downstream target of the male sex determining gene SRY. AB - The cascade of molecular events involved in mammalian sex determination has been shown to involve the SRY gene, but specific downstream events have eluded researchers for decades. The current study identifies one of the first direct downstream targets of the male sex determining factor SRY as the basic-helix-loop helix (bHLH) transcription factor TCF21. SRY was found to bind to the Tcf21 promoter and activate gene expression. Mutagenesis of SRY/SOX9 response elements in the Tcf21 promoter eliminated the actions of SRY. SRY was found to directly associate with the Tcf21 promoter SRY/SOX9 response elements in vivo during fetal rat testis development. TCF21 was found to promote an in vitro sex reversal of embryonic ovarian cells to induce precursor Sertoli cell differentiation. TCF21 and SRY had similar effects on the in vitro sex reversal gonadal cell transcriptomes. Therefore, SRY acts directly on the Tcf21 promoter to in part initiate a cascade of events associated with Sertoli cell differentiation and embryonic testis development. PMID- 21637324 TI - Review of cardiotoxicity in pediatric cancer patients: during and after therapy. AB - With the improvement in survival from childhood cancer, late effects of therapy are becoming more apparent. Cardiac disease, one of these late effects, has a significant impact on the life of survivors of childhood cancers. Most survivors are followed by primary care doctors and adult subspecialists after they have graduated from pediatric centers. Since much of the cardiac toxicity of therapy occurs years off of therapy, it is important for these physicians to be aware of how to monitor survivors for the development of cardiac toxicities. In this paper we will discuss the incidence of cardiac disease during treatment and in survivors, what treatment modalities contribute to its development and modalities utilized to screen for cardiac disease. Recommendations for posttherapy monitoring will be emphasized. PMID- 21637325 TI - Enhancement of Cellulase Activity from a New Strain of Bacillus subtilis by Medium Optimization and Analysis with Various Cellulosic Substrates. AB - The cellulase activity of Bacillus subtilis AS3 was enhanced by optimizing the medium composition by statistical methods. The enzyme activity with unoptimised medium with carboxymethylcellulose (CMC) was 0.07 U/mL and that was significantly enhanced by CMC, peptone, and yeast extract using Placket-Burman design. The combined effects of these nutrients on cellulase activity were studied using 2(2) full factorial central composite design. The optimal levels of medium components determined were CMC (1.8%), peptone (0.8%), and yeast extract (0.479%). The maximum enzyme activity predicted by the model was 0.49 U/mL which was in good agreement with the experimental value 0.43 U/mL showing 6-fold increase as compared to unoptimised medium. The enzyme showed multisubstrate specificity, showing significantly higher activity with lichenan and beta-glucan and lower activity with laminarin, hydroxyethylcellulose, and steam exploded bagasse. The optimised medium with lichenan or beta-glucan showed 2.5- or 2.8-fold higher activity, respectively, at same concentration as of CMC. PMID- 21637326 TI - Identification of a Functional Type IA Topoisomerase, LdTopIIIbeta, from Kinetoplastid Parasite Leishmania donovani. AB - DNA topoisomerases of kinetoplastids represent a family of DNA processing enzymes that essentially solve the topological problems not only in nuclear DNA but also in kinetoplast DNA. We have, for the first time, identified a Leishmania donovani homologue of bacterial and eukaryotic IA type of topoisomerase III protein and termed as LdTopIIIbeta. Complementation study of wild-type and mutant LdTopIIIbeta with slow-growing topoisomerase III mutant yeast S. cerevisiae revealed the functional conservation of the leishmanial counterpart of topoisomerase IIIbeta protein, the 327 tyrosine being the active site amino acid. A C-terminal deletion construct of LdTopIIIbeta could not suppress the slow growth phenotype of mutant yeast, indicating the requirement of C-terminal region for the enzyme function in vivo.LdTopIIIbeta localized inside the nucleus and kinetoplast of the parasite. Taken together, our study indicates functional conservation and possible role of LdTopIIIbeta in parasite DNA processing. PMID- 21637327 TI - Inhibitory effect of phthalic Acid on tyrosinase: the mixed-type inhibition and docking simulations. AB - Tyrosinase inhibition studies are needed due to the medicinal applications such as hyperpigmentation. For probing effective inhibitors of tyrosinase, a combination of computational prediction and enzymatic assay via kinetics was important. We predicted the 3D structure of tyrosinase, used a docking algorithm to simulate binding between tyrosinase and phthalic acid (PA), and studied the reversible inhibition of tyrosinase by PA. PA inhibited tyrosinase in a mixed type manner with a K(i) = 65.84 +/- 1.10 mM. Measurements of intrinsic and ANS binding fluorescences showed that PA induced changes in the active site structure via indirect binding. Simulation was successful (binding energies for Dock6.3 = 27.22 and AutoDock4.2 = -0.97 kcal/mol), suggesting that PA interacts with LEU73 residue that is predicted commonly by both programs. The present study suggested that the strategy of predicting tyrosinase inhibition based on hydroxyl groups and orientation may prove useful for screening of potential tyrosinase inhibitors. PMID- 21637328 TI - Evaluation of Chlorella (Chlorophyta) as Source of Fermentable Sugars via Cell Wall Enzymatic Hydrolysis. AB - The cell wall of Chlorella is composed of up to 80% carbohydrates including cellulose. In this study, Chlorella homosphaera and Chlorella zofingiensis were evaluated as source of fermentable sugars via their cell wall enzymatic degradation. The algae were cultivated in inorganic medium, collected at the stationary growth phase and centrifuged. The cell pellet was suspended in citrate buffer, pH 4.8 and subjected to 24 hours hydrolysis at 50 degrees C using a cellulases, xylanases, and amylases blend. The measurement of glucose and reducing sugars concentration in the reaction mixture supernatant, on a dry biomass base, showed hydrolysis yields of 2.9% and 5.03% glucose and 4.8% and 8.6% reducing sugars, for C. homosphaera and C. zofingiensis, respectively. However if cells were washed with chilled ethanol, cold dried, and grounded the biomass hydrolysis yields increased to 23.3% and 18.4% glucose and 24.5% and 19.3% reducing sugars for C. homosphaera and C. zofingiensis, respectively. PMID- 21637329 TI - Chemistry based on renewable raw materials: perspectives for a sugar cane-based biorefinery. AB - Carbohydrates are nowadays a very competitive feedstock for the chemical industry because their availability is compatible with world-scale chemical production and their price, based on the carbon content, is comparable to that of petrochemicals. At the same time, demand is rising for biobased products. Brazilian sugar cane is a competitive feedstock source that is opening the door to a wide range of bio-based products. This essay begins with the importance of the feedstock for the chemical industry and discusses developments in sugar cane processing that lead to low cost feedstocks. Thus, sugar cane enables a new chemical industry, as it delivers a competitive raw material and a source of energy. As a result, sugar mills are being transformed into sustainable biorefineries that fully exploit the potential of sugar cane. PMID- 21637330 TI - Performance and consistency of indicator groups in two biodiversity hotspots. AB - BACKGROUND: In a world limited by data availability and limited funds for conservation, scientists and practitioners must use indicator groups to define spatial conservation priorities. Several studies have evaluated the effectiveness of indicator groups, but still little is known about the consistency in performance of these groups in different regions, which would allow their a priori selection. METHODOLOGY/PRINCIPAL FINDINGS: We systematically examined the effectiveness and the consistency of nine indicator groups in representing mammal species in two top-ranked Biodiversity Hotspots (BH): the Brazilian Cerrado and the Atlantic Forest. To test for group effectiveness we first found the best sets of sites able to maximize the representation of each indicator group in the BH and then calculated the average representation of different target species by the indicator groups in the BH. We considered consistent indicator groups whose representation of target species was not statistically different between BH. We called effective those groups that outperformed the target-species representation achieved by random sets of species. Effective indicator groups required the selection of less than 2% of the BH area for representing target species. Restricted-range species were the most effective indicators for the representation of all mammal diversity as well as target species. It was also the only group with high consistency. CONCLUSIONS/SIGNIFICANCE: We show that several indicator groups could be applied as shortcuts for representing mammal species in the Cerrado and the Atlantic Forest to develop conservation plans, however, only restricted-range species consistently held as the most effective indicator group for such a task. This group is of particular importance in conservation planning as it captures high diversity of endemic and endangered species. PMID- 21637331 TI - Estimating parameters of speciation models based on refined summaries of the joint site-frequency spectrum. AB - Understanding the processes and conditions under which populations diverge to give rise to distinct species is a central question in evolutionary biology. Since recently diverged populations have high levels of shared polymorphisms, it is challenging to distinguish between recent divergence with no (or very low) inter-population gene flow and older splitting events with subsequent gene flow. Recently published methods to infer speciation parameters under the isolation migration framework are based on summarizing polymorphism data at multiple loci in two species using the joint site-frequency spectrum (JSFS). We have developed two improvements of these methods based on a more extensive use of the JSFS classes of polymorphisms for species with high intra-locus recombination rates. First, using a likelihood based method, we demonstrate that taking into account low-frequency polymorphisms shared between species significantly improves the joint estimation of the divergence time and gene flow between species. Second, we introduce a local linear regression algorithm that considerably reduces the computational time and allows for the estimation of unequal rates of gene flow between species. We also investigate which summary statistics from the JSFS allow the greatest estimation accuracy for divergence time and migration rates for low (around 10) and high (around 100) numbers of loci. Focusing on cases with low numbers of loci and high intra-locus recombination rates we show that our methods for the estimation of divergence time and migration rates are more precise than existing approaches. PMID- 21637332 TI - Tim-3 negatively regulates IL-12 expression by monocytes in HCV infection. AB - T cell immunoglobulin and mucin domain-containing protein 3 (Tim-3) is a newly identified negative immunomodulator that is up-regulated on dysfunctional T cells during viral infections. The expression and function of Tim-3 on human innate immune responses during HCV infection, however, remains poorly characterized. In this study, we report that Tim-3 is constitutively expressed on human resting CD14(+) monocyte/macrophages (M/M(O)) and functions as a cap to block IL-12, a key pro-inflammatory cytokine linking innate and adaptive immune responses. Tim-3 expression is significantly reduced and IL-12 expression increased upon stimulation with Toll-like receptor 4 (TLR4) ligand--lipopolysaccharide (LPS) and TLR7/8 ligand--R848. Notably, Tim-3 is over-expressed on un-stimulated as well as TLR-stimulated M/M(O), which is inversely associated with the diminished IL-12 expression in chronically HCV-infected individuals when compared to healthy subjects. Up-regulation of Tim-3 and inhibition of IL-12 are also observed in M/M(O) incubated with HCV-expressing hepatocytes, as well as in primary M/M(O) or monocytic THP-1 cells incubated with HCV core protein, an effect that mimics the function of complement C1q and is reversible by blocking the HCV core/gC1qR interaction. Importantly, blockade of Tim-3 signaling significantly rescues HCV mediated inhibition of IL-12, which is primarily expressed by Tim-3 negative M/M(O). Tim-3 blockade reduces HCV core-mediated expression of the negative immunoregulators PD-1 and SOCS-1 and increases STAT-1 phosphorylation. Conversely, blocking PD-1 or silencing SOCS-1 gene expression also decreases Tim 3 expression and enhances IL-12 secretion and STAT-1 phosphorylation. These findings suggest that Tim-3 plays a crucial role in negative regulation of innate immune responses, through crosstalk with PD-1 and SOCS-1 and limiting STAT-1 phosphorylation, and may be a novel target for immunotherapy to HCV infection. PMID- 21637333 TI - Cost-effectiveness of preoperative screening and eradication of Staphylococcus aureus carriage. AB - BACKGROUND: Preoperative screening for nasal S. aureus carriage, followed by eradication treatment of identified carriers with nasal mupirocine ointment and chlorhexidine soap was highly effective in preventing deep-seated S. aureus infections. It is unknown how cost-effectiveness of this intervention is affected by suboptimal S. aureus screening. We determined cost-effectiveness of different preoperative S. aureus screening regimes. METHODS: We compared different screening scenarios (ranging from treating all patients without screening to treating only identified S. aureus carriers) to the base case scenario without any screening and treatment. Screening and treatment costs as well as costs and mortality due to deep-seated S. aureus infection were derived from hospital databases and prospectively collected data, respectively. RESULTS: As compared to the base case scenario, all scenarios are associated with improved health care outcomes at reduced costs. Treating all patients without screening is most cost beneficial, saving ?7339 per life year gained, as compared to ?3330 when only identified carriers are treated. In sensitivity analysis, outcomes are susceptible to the sensitivity of the screening test and the efficacy of treatment. Reductions in these parameters would reduce the cost-effectiveness of scenarios in which treatment is based on screening. When only identified S. aureus carriers are treated costs of screening should be less than ?6.23 to become the dominant strategy. CONCLUSIONS: Preoperative screening and eradication of S. aureus carriage to prevent deep-seated S. aureus infections saves both life years and medical costs at the same time, although treating all patients without screening is the dominant strategy, resulting in most health gains and largest savings. PMID- 21637334 TI - The characterisation of three types of genes that overlie copy number variable regions. AB - BACKGROUND: Due to the increased accuracy of Copy Number Variable region (CNV) break point mapping, it is now possible to say with a reasonable degree of confidence whether a gene (i) falls entirely within a CNV; (ii) overlaps the CNV or (iii) actually contains the CNV. We classify these as type I, II and III CNV genes respectively. PRINCIPAL FINDINGS: Here we show that although type I genes vary in copy number along with the CNV, most of these type I genes have the same expression levels as wild type copy numbers of the gene. These genes must, therefore, be under homeostatic dosage compensation control. Looking into possible mechanisms for the regulation of gene expression we found that type I genes have a significant paucity of genes regulated by miRNAs and are not significantly enriched for monoallelically expressed genes. Type III genes, on the other hand, have a significant excess of genes regulated by miRNAs and are enriched for genes that are monoallelically expressed. SIGNIFICANCE: Many diseases and genomic disorders are associated with CNVs so a better understanding of the different ways genes are associated with normal CNVs will help focus on candidate genes in genome wide association studies. PMID- 21637335 TI - Multiple-locus variable number tandem repeat analysis for Streptococcus pneumoniae: comparison with PFGE and MLST. AB - In the era of pneumococcal conjugate vaccines, surveillance of pneumococcal disease and carriage remains of utmost importance as important changes may occur in the population. To monitor these alterations reliable genotyping methods are required for large-scale applications. We introduced a high throughput multiple locus variable number tandem repeat analysis (MLVA) and compared this method with pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). The MLVA described here is based on 8 BOX loci that are amplified in two multiplex PCRs. The labeled PCR products are sized on an automated DNA sequencer to accurately determine the number of tandem repeats. The composite of the number of repeats of the BOX loci makes up a numerical profile that is used for identification and clustering. In this study, MLVA was performed on 263 carriage isolates that were previously characterized by MLST and PFGE. MLVA, MLST and PFGE (cut-off of 80%) yielded 164, 120, and 87 types, respectively. The three typing methods had Simpson's diversity indices of 98.5% or higher. Congruence between MLST and MLVA was high. The Wallace of MLVA to MLST was 0.874, meaning that if two strains had the same MLVA type they had an 88% chance of having the same MLST type. Furthermore, the Wallace of MLVA to clonal complex of MLST was even higher: 99.5%. For some isolates belonging to a single MLST clonal complex although displaying different serotypes, MLVA was more discriminatory, generating groups according to serotype or serogroup. Overall, MLVA is a promising genotyping method that is easy to perform and a relatively cheap alternative to PFGE and MLST. In the companion paper published simultaneously in this issue we applied the MLVA to assess the pneumococcal population structure of isolates causing invasive disease in The Netherlands before the introduction of the 7-valent conjugate vaccine. PMID- 21637336 TI - Choosing and using a plant DNA barcode. AB - The main aim of DNA barcoding is to establish a shared community resource of DNA sequences that can be used for organismal identification and taxonomic clarification. This approach was successfully pioneered in animals using a portion of the cytochrome oxidase 1 (CO1) mitochondrial gene. In plants, establishing a standardized DNA barcoding system has been more challenging. In this paper, we review the process of selecting and refining a plant barcode; evaluate the factors which influence the discriminatory power of the approach; describe some early applications of plant barcoding and summarise major emerging projects; and outline tool development that will be necessary for plant DNA barcoding to advance. PMID- 21637337 TI - Field testing of different chemical combinations as odour baits for trapping wild mosquitoes in The Gambia. AB - Odour baited traps have potential use in population surveillance of insect vectors of disease, and in some cases for vector population reduction. Established attractants for human host-seeking mosquitoes include a combination of CO(2) with L-lactic acid and ammonia, on top of which additional candidate compounds are being tested. In this field study in rural Gambia, using Latin square experiments with thorough randomization and replication, we tested nine different leading candidate combinations of chemical odorants for attractiveness to wild mosquitoes including anthropophilic malaria vectors, using modified Mosquito Magnet-X (MM-X) counterflow traps outside experimental huts containing male human sleepers. Highest catches of female mosquitoes, particularly of An. gambiae s.l. and Mansonia species, were obtained by incorporation of tetradecanoic acid. As additional carboxylic acids did not increase the trap catches further, this 'reference blend' (tetradecanoic acid with L-lactic acid, ammonia and CO(2)) was used in subsequent experiments. MM-X traps with this blend caught similar numbers of An. gambiae s.l. and slightly more Mansonia and Culex mosquitoes than a standard CDC light trap, and these numbers were not significantly affected by the presence or absence of human sleepers in the huts. Experiments with CO(2) produced from overnight yeast cultures showed that this organic source was effective in enabling trap attractiveness for all mosquito species, although at a slightly lower efficiency than obtained with use of CO(2) gas cylinders. Although further studies are needed to discover additional chemicals that increase attractiveness, as well as to optimise trap design and CO(2) source for broader practical use, the odour-baited traps described here are safe and effective for sampling host-seeking mosquitoes outdoors and can be incorporated into studies of malaria vector ecology. PMID- 21637338 TI - Serine/threonine protein kinase SpkG is a candidate for high salt resistance in the unicellular cyanobacterium Synechocystis sp. PCC 6803. AB - BACKGROUND: Seven serine/threonine kinase genes have been predicted in unicellular cyanobacterium Synechocystis sp. PCC6803. SpkA and SpkB were shown to be required for cell motility and SpkE has no kinase activity. There is no report whether the other four STKs are involved in stress-mediated signaling in Synechocystis PCC6803. METHODOLOGY/PRINCIPAL FINDINGS: In this paper, we examined differential expression of the other four serine/threonine kinases, SpkC, SpkD, SpkF and SpkG, at seven different stress conditions. The transcriptional level was up-regulated of spkG and down-regulated of spkC under high salt stress condition. Two spk deletion mutants, DeltaspkC and DeltaspkG, were constructed and their growth characteristic were examined compared to the wild strain. The wild strain and DeltaspkC mutant were not affected under high salt stress conditions. In contrast, growth of spkG mutant was completely impaired. To further confirm the function of spkG, we also examined the effect of mutation of spkG on the expression of salt stress-inducible genes. We compared genome-wide patterns of transcription between wild-type Synechocystis sp. PCC6803 and cells with a mutation in the SpkG with DNA microarray analysis. CONCLUSION: In this study, we first study the spkG gene as sensor of high salt signal. We consider that SpkG play essential roles in Synechocystis sp. for sensing the high salt signal directly, rather than mediating signals among other kinases. Our microarray experiment may help select relatively significant genes for further research on mechanisms of signal transduction of Synechocystis sp. PCC6803 under high salt stress. PMID- 21637339 TI - Cell entry and trafficking of human adenovirus bound to blood factor X is determined by the fiber serotype and not hexon:heparan sulfate interaction. AB - Human adenovirus serotype 5 (HAdV5)-based vectors administered intravenously accumulate in the liver as the result of their direct binding to blood coagulation factor X (FX) and subsequent interaction of the FX-HAdV5 complex with heparan sulfate proteoglycan (HSPG) at the surface of liver cells. Intriguingly, the serotype 35 fiber-pseudotyped vector HAdV5F35 has liver transduction efficiencies 4-logs lower than HAdV5, even though both vectors carry the same hexon capsomeres. In order to reconcile this apparent paradox, we investigated the possible role of other viral capsid proteins on the FX/HSPG-mediated cellular uptake of HAdV5-based vectors. Using CAR- and CD46-negative CHO cells varying in HSPG expression, we confirmed that FX bound to serotype 5 hexon protein and to HAdV5 and HAdV5F35 virions via its Gla-domain, and enhanced the binding of both vectors to surface-immobilized hypersulfated heparin and cellular HSPG. Using penton mutants, we found that the positive effect of FX on HAdV5 binding to HSPG and cell transduction did not depend on the penton base RGD and fiber shaft KKTK motifs. However, we found that FX had no enhancing effect on the HAdV5F35 mediated cell transduction, but a negative effect which did not involve the cell attachment or endocytic step, but the intracellular trafficking and nuclear import of the FX-HAdV5F35 complex. By cellular imaging, HAdV5F35 particles were observed to accumulate in the late endosomal compartment, and were released in significant amounts into the extracellular medium via exocytosis. We showed that the stability of serotype 5 hexon:FX interaction was higher at low pH compared to neutral pH, which could account for the retention of FX-HAdV5F35 complexes in the late endosomes. Our results suggested that, despite the high affinity interaction of hexon capsomeres to FX and cell surface HSPG, the adenoviral fiber acted as the dominant determinant of the internalization and trafficking pathway of HAdV5 based vectors. PMID- 21637340 TI - Evaluation of magnetic micro- and nanoparticle toxicity to ocular tissues. AB - PURPOSE: Magnetic nanoparticles (MNPs) may be used for focal delivery of plasmids, drugs, cells, and other applications. Here we ask whether such particles are toxic to ocular structures. METHODS: To evaluate the ocular toxicity of MNPs, we asked if either 50 nm or 4 um magnetic particles affect intraocular pressure, corneal endothelial cell count, retinal morphology including both cell counts and glial activation, or photoreceptor function at different time points after injection. Sprague-Dawley rats (n = 44) were injected in the left eye with either 50 nm (3 ul, 1.65 mg) or 4 um (3 ul, 1.69 mg) magnetic particles, and an equal volume of PBS into the right eye. Electroretinograms (ERG) were used to determine if MNPs induce functional changes to the photoreceptor layers. Enucleated eyes were sectioned for histology and immunofluorescence. RESULTS: Compared to control-injected eyes, MNPs did not alter IOP measurements. ERG amplitudes for a-waves were in the 100-250 uV range and b-waves were in the 500-600 uV range, with no significant differences between injected and non-injected eyes. Histological sectioning and immunofluorescence staining showed little difference in MNP-injected animals compared to control eyes. In contrast, at 1 week, corneal endothelial cell numbers were significantly lower in the 4 um magnetic particle-injected eyes compared to either 50 nm MNP- or PBS-injected eyes. Furthermore, iron deposition was detected after 4 um magnetic particle but not 50 nm MNP injection. CONCLUSIONS: Intravitreal or anterior chamber injections of MNPs showed little to no signs of toxicity on retinal structure, photoreceptor function or aqueous drainage in the eye. Our results suggest that MNPs are safe for intraocular use. PMID- 21637341 TI - Allele-specific, age-dependent and BMI-associated DNA methylation of human MCHR1. AB - BACKGROUND: Melanin-concentrating hormone receptor 1 (MCHR1) plays a significant role in regulation of energy balance, food intake, physical activity and body weight in humans and rodents. Several association studies for human obesity showed contrary results concerning the SNPs rs133072 (G/A) and rs133073 (T/C), which localize to the first exon of MCHR1. The variations constitute two main haplotypes (GT, AC). Both SNPs affect CpG dinucleotides, whereby each haplotype contains a potential methylation site at one of the two SNP positions. In addition, 15 CpGs in close vicinity of these SNPs constitute a weak CpG island. Here, we studied whether DNA methylation in this sequence context may contribute to population- and age-specific effects of MCHR1 alleles in obesity. PRINCIPAL FINDINGS: We analyzed DNA methylation of a 315 bp region of MCHR1 encompassing rs133072 and rs133073 and the CpG island in blood samples of 49 individuals by bisulfite sequencing. The AC haplotype shows a significantly higher methylation level than the GT haplotype. This allele-specific methylation is age-dependent. In young individuals (20-30 years) the difference in DNA methylation between haplotypes is significant; whereas in individuals older than 60 years it is not detectable. Interestingly, the GT allele shows a decrease in methylation status with increasing BMI, whereas the methylation of the AC allele is not associated with this phenotype. Heterozygous lymphoblastoid cell lines show the same pattern of allele-specific DNA methylation. The cell line, which exhibits the highest difference in methylation levels between both haplotypes, also shows allele specific transcription of MCHR1, which can be abolished by treatment with the DNA methylase inhibitor 5-aza-2'-deoxycytidine. CONCLUSIONS: We show that DNA methylation at MCHR1 is allele-specific, age-dependent, BMI-associated and affects transcription. Conceivably, this epigenetic regulation contributes to the age- and/or population specific effects reported for MCHR1 in several human obesity studies. PMID- 21637342 TI - Early estimation of the reproduction number in the presence of imported cases: pandemic influenza H1N1-2009 in New Zealand. AB - We analyse data from the early epidemic of H1N1-2009 in New Zealand, and estimate the reproduction number R. We employ a renewal process which accounts for imported cases, illustrate some technical pitfalls, and propose a novel estimation method to address these pitfalls. Explicitly accounting for the infection-age distribution of imported cases and for the delay in transmission dynamics due to international travel, R was estimated to be (95% confidence interval: 107,1.47). Hence we show that a previous study, which did not account for these factors, overestimated R. Our approach also permitted us to examine the infection-age at which secondary transmission occurs as a function of calendar time, demonstrating the downward bias during the beginning of the epidemic. These technical issues may compromise the usefulness of a well-known estimator of R- the inverse of the moment-generating function of the generation time given the intrinsic growth rate. Explicit modelling of the infection-age distribution among imported cases and the examination of the time dependency of the generation time play key roles in avoiding a biased estimate of R, especially when one only has data covering a short time interval during the early growth phase of the epidemic. PMID- 21637343 TI - Vascular endothelial growth factor in the circulation in cancer patients may not be a relevant biomarker. AB - BACKGROUND: Levels of circulating vascular endothelial growth factor (VEGF) have widely been used as biomarker for angiogenic activity in cancer. For this purpose, non-standardized measurements in plasma and serum were used, without correction for artificial VEGF release by platelets activated ex vivo. We hypothesize that "true" circulating (c)VEGF levels in most cancer patients are low and unrelated to cancer load or tumour angiogenesis. METHODOLOGY: We determined VEGF levels in PECT, a medium that contains platelet activation inhibitors, in citrate plasma, and in isolated platelets in 16 healthy subjects, 18 patients with metastatic non-renal cancer (non-RCC) and 12 patients with renal cell carcinoma (RCC). In non-RCC patients, circulating plasma VEGF levels were low and similar to VEGF levels in controls if platelet activation was minimized during the harvest procedure by PECT medium. In citrate plasma, VEGF levels were elevated in non-RCC patients, but this could be explained by a combination of increased platelet activation during blood harvesting, and by a two-fold increase in VEGF content of individual platelets (controls: 3.4 IU/10(6), non-RCC: 6.2 IU/10(6) platelets, p = 0.001). In contrast, cVEGF levels in RCC patients were elevated (PECT plasma: 64 pg/ml vs. 21 pg/ml, RCC vs. non-RCC, p<0.0001), and not related to platelet VEGF concentration. CONCLUSIONS: Our findings suggest that "true" freely cVEGF levels are not elevated in the majority of cancer patients. Previously reported elevated plasma VEGF levels in cancer appear to be due to artificial release from activated platelets, which in cancer have an increased VEGF content, during the blood harvest procedure. Only in patients with RCC, which is characterized by excessive VEGF production due to a specific genetic defect, were cVEGF levels elevated. This observation may be related to limited and selective success of anti-VEGF agents, such as bevacizumab and sorafenib, as monotherapy in RCC compared to other forms of cancer. PMID- 21637344 TI - Cue integration in categorical tasks: insights from audio-visual speech perception. AB - Previous cue integration studies have examined continuous perceptual dimensions (e.g., size) and have shown that human cue integration is well described by a normative model in which cues are weighted in proportion to their sensory reliability, as estimated from single-cue performance. However, this normative model may not be applicable to categorical perceptual dimensions (e.g., phonemes). In tasks defined over categorical perceptual dimensions, optimal cue weights should depend not only on the sensory variance affecting the perception of each cue but also on the environmental variance inherent in each task-relevant category. Here, we present a computational and experimental investigation of cue integration in a categorical audio-visual (articulatory) speech perception task. Our results show that human performance during audio-visual phonemic labeling is qualitatively consistent with the behavior of a Bayes-optimal observer. Specifically, we show that the participants in our task are sensitive, on a trial by-trial basis, to the sensory uncertainty associated with the auditory and visual cues, during phonemic categorization. In addition, we show that while sensory uncertainty is a significant factor in determining cue weights, it is not the only one and participants' performance is consistent with an optimal model in which environmental, within category variability also plays a role in determining cue weights. Furthermore, we show that in our task, the sensory variability affecting the visual modality during cue-combination is not well estimated from single-cue performance, but can be estimated from multi-cue performance. The findings and computational principles described here represent a principled first step towards characterizing the mechanisms underlying human cue integration in categorical tasks. PMID- 21637345 TI - A Sir2-like protein participates in mycobacterial NHEJ. AB - In eukaryotic cells, repair of DNA double-strand breaks (DSBs) by the nonhomologous end-joining (NHEJ) pathway is critical for genome stability. In contrast to the complex eukaryotic repair system, bacterial NHEJ apparatus consists of only two proteins, Ku and a multifunctional DNA ligase (LigD), whose functional mechanism has not been fully clarified. We show here for the first time that Sir2 is involved in the mycobacterial NHEJ repair pathway. Here, using tandem affinity purification (TAP) screening, we have identified an NAD-dependent deacetylase in mycobacteria which is a homologue of the eukaryotic Sir2 protein and interacts directly with Ku. Results from an in vitro glutathione S transferase (GST) pull-down assay suggest that Sir2 interacts directly with LigD. Plasmid-based end-joining assays revealed that the efficiency of DSB repair in a sir2 deletion mutant was reduced 2-fold. Moreover, the Deltasir2 strain was about 10-fold more sensitive to ionizing radiation (IR) in the stationary phase than the wild-type. Our results suggest that Sir2 may function closely together with Ku and LigD in the nonhomologous end-joining pathway in mycobacteria. PMID- 21637346 TI - Role of IL-1 beta in the development of human T(H)17 cells: lesson from NLPR3 mutated patients. AB - BACKGROUND: T helper 17 cells (T(H)-17) represent a lineage of effector T cells critical in host defence and autoimmunity. In both mouse and human IL-1beta has been indicated as a key cytokine for the commitment to T(H)-17 cells. Cryopyrin associated periodic syndromes (CAPS) are a group of inflammatory diseases associated with mutations of the NLRP3 gene encoding the inflammasome component cryopyrin. In this work we asked whether the deregulated secretion of IL-1beta secondary to mutations characterizing these patients could affect the IL-23/IL-17 axis. METHODOLOGY/PRINCIPAL FINDINGS: A total of 11 CAPS, 26 systemic onset juvenile idiopathic arthritis (SoJIA) patients and 20 healthy controls were analyzed. Serum levels of IL-17 and IL-6 serum were assessed by ELISA assay. Frequency of T(H)17 cells was quantified upon staphylococcus enterotoxin B (SEB) stimulation. Secretion of IL-1beta, IL-23 and IL-6 by monocyte derived dendritic cells (MoDCs), were quantified by ELISA assay. A total of 8 CAPS and 11 SoJIA patients were also analysed before and after treatment with IL-1beta blockade. Untreated CAPS patients showed significantly increased IL-17 serum levels as well as a higher frequency of T(H)17 compared to control subjects. On the contrary, SoJIA patients displayed a frequency of T(H)17 similar to normal donors, but were found to have significantly increased serum level of IL-6 when compared to CAPS patients or healthy donors. Remarkably, decreased IL-17 serum levels and T(H)17 frequency were observed in CAPS patients following in vivo IL-1beta blockade. On the same line, MoDCs from CAPS patients exhibited enhanced secretion of IL-1beta and IL-23 upon TLRs stimulation, with a reduction after anti-IL-1 treatment. CONCLUSION/SIGNIFICANCE: These findings further support the central role of IL 1beta in the differentiation of T(H)17 in human inflammatory conditions. PMID- 21637347 TI - Prevalence and risk factors for swine influenza virus infection in the English pig population. AB - Infection of pigs with influenza viruses is a cause of considerable economic loss for pig farmers as well as a potential human health concern - as evidenced by the identification of genetic material derived from swine-adapted influenza viruses in an novel strain of H1N1 influenza virus in 2009. A study was conducted investigating the prevalence of influenza virus infection in a selection of 143 English pig herds between April 2008 and April 2009, which found evidence of recent virus circulation in over half of these herds (n=75). Farms which were sampled in the Summer months were found to have lower odds of recent virus circulation, as were farms containing pigs kept in straw yards. Additionally, farms containing pigs kept indoors and farms containing high numbers of finisher pigs per water space were found to have higher odds of recent virus circulation. It is hoped that further studies will expand on these findings, and may allow targeting of surveillance for influenza viruses in the English pig population. PMID- 21637348 TI - HIV Disease Impact on Mothers: What They Miss During Their Children's Developmental Years. AB - Adjusting to chronic illness is very complicated for families with children, as they are already faced with the challenge of development and childrearing. In this study, qualitative interviews were conducted with HIV positive mothers on a number of issues related to being an HIV positive mother raising young children. One topic of the interview was whether or not they felt that HIV had caused them to miss activities with their children while the children were growing up, what types of activities they had missed, the age of the child for each example, and how HIV had led to missing these activities. Interviews were conducted in 2008 with a random sample of 57 mothers being followed in a longitudinal assessment study. All study participants were English or Spanish speaking. Mean age was 44.1 (SD = 5.6) years; 47% were Latina; 35% African American; 11% White; and 7% other race. About 60% of the mothers disclosed that their HIV status had caused them to miss out on activities with their children while their children were growing up, ranging from daily care activities to major school and extra-curricular activities. Some mothers missed significant amounts of time with their children due to hospitalizations. In some cases mothers felt forced into a choice between mothering ability and their own health, including adherence to medications. Implications for the mothers and the children are discussed. PMID- 21637349 TI - Drawing and interpreting data: Children's impressions of onchocerciasis and community-directed treatment with ivermectin (CDTI) in four onchocerciasis endemic countries in Africa. AB - Although the depiction of a child leading a blind man is the most enduring image of onchocerciasis in Africa, research activities have hardly involved children. This paper aims at giving voice to children through drawings and their interpretation. The study was conducted in 2009 in Cameroon, Democratic Republic of Congo (DRC), Nigeria and Uganda. Children aged 6-16 years were asked to draw their perceptions of onchocerciasis and community-directed treatment with ivermectin (CDTI) in their communities. A total of 50 drawings were generated. The drawings depicted four main aspects of onchocerciasis: (1) the disease symptoms, (2) the negative consequences of onchocerciasis among children and in the community generally, (3) the ivermectin distribution process, and (4) the benefits or effects of taking ivermectin. Out of the 50 drawings, 30 were on symptoms, 7 on effects of the disease on children, 8 on distribution process, and 5 represented multiple perceptions on symptoms, drug distribution processes, benefits, and effects of treatment. The lack of clarity when treatment with ivermectin can be stopped in endemic areas requires working with children to ensure continued compliance with treatment into the future. Children's drawings should be incorporated into health education interventions. PMID- 21637350 TI - Maintaining families' well-being in everyday life. AB - The aim of this paper is to discuss how everyday life changes for the family in the event of chronic illness or disability. It changes physically due to loss of body function and socially due to time and other constraints related to treatment or lack of mobility. Equally important, there is a psychological impact due to the uncertainty of the future. The article will explore how family participation can help to maintain well-being in everyday life. The family should therefore focus on their own needs as much as on the needs of the family members who are ill. In order to maintain well-being in everyday life, it is crucial for the family to create routines and spend time doing things that they enjoy. By doing this, the family will create a rhythm of well-being regardless of the critical family situation. Family members and professional caregivers also need to come together at the beginning and during the illness or disability event to discuss changes that could be made day-to-day for all those involved, thereby making for an easier transition into care giving. PMID- 21637351 TI - How a mycoparasite employs g-protein signaling: using the example of trichoderma. AB - Mycoparasitic Trichoderma spp. act as potent biocontrol agents against a number of plant pathogenic fungi, whereupon the mycoparasitic attack includes host recognition followed by infection structure formation and secretion of lytic enzymes and antifungal metabolites leading to the host's death. Host-derived signals are suggested to be recognized by receptors located on the mycoparasite's cell surface eliciting an internal signal transduction cascade which results in the transcription of mycoparasitism-relevant genes. Heterotrimeric G proteins of fungi transmit signals originating from G-protein-coupled receptors mainly to the cAMP and the MAP kinase pathways resulting in regulation of downstream effectors. Components of the G-protein signaling machinery such as Galpha subunits and G protein-coupled receptors were recently shown to play crucial roles in Trichoderma mycoparasitism as they govern processes such as the production of extracellular cell wall lytic enzymes, the secretion of antifungal metabolites, and the formation of infection structures. PMID- 21637352 TI - GIMAP Proteins in T-Lymphocytes. AB - (GIMAPs) GTPase of the immunity associated protein family are a novel protein family of putative small GTPases. GIMAPs are mainly expressed in the cells of the immune system and have been associated with immunological functions, such as thymocyte development, apoptosis of peripheral lymphocytes and T helper cell differentiation. GIMAPs have also been linked to immunological diseases, such as T cell lymphopenia, leukemia and autoimmune diseases. In this review we examine the role of GIMAP proteins in T-lymphocyte biology. PMID- 21637353 TI - Posttranslational processing and modification of cathepsins and cystatins. AB - Cathepsins are an essential protease family in all living cells. The cathepsins play an essential roles such as protein catabolism and protein synthesis. To targeting to various organella and to regulate their activity, the post translational-processing and modification play an important role Cathepsins are translated in polysome as the pre-pro-mature forms. The pre-peptide is removed cotranslationally and then translocated to Golgi-apparatus and the pro-part is removed and the mature-part is glycosylated, and the mature-part is targeted into the lysosome mediated by mannose-6-phosphate signal and the mature-part is bound with their coenzymes. The degradation of the mature-part is started by the limited proteolysis of the ordered nicked bonds to make hydrophobic peptides. The peptides are incorporated into phagosome or proteasome after ubiquitinated and are degrade into amino-acids. Cystatins are endogenous inhibitors of cathepsins. Cystatin alpha which is only located in skin is phosphorylated at the near C terminus by protein kinase-C, and the phosphorylate-cystatin alpha is incorporated into cornified envelope and conjugated with filaggrin-fiber by transglutaminase to form the linker-fiber of skin. The cystatin alpha is modified by glutathione or make their dimmer, and they are inactive. Those modifications are regulated by the redox-potential by the glutathione. PMID- 21637354 TI - Suppression by Ghrelin of Porphyromonas gingivalis-Induced Constitutive Nitric Oxide Synthase S-Nitrosylation and Apoptosis in Salivary Gland Acinar Cells. AB - Oral mucosal inflammatory responses to periodontopathic bacterium, P. gingivalis, and its key virulence factor, LPS, are characterized by a massive rise in epithelial cell apoptosis and the disturbances in NO signaling pathways. Here, we report that the LPS-induced enhancement in rat sublingual salivary gland acinar cell apoptosis and NO generation was associated with the suppression in constitutive nitric oxide synthase (cNOS) activity and a marked increase in the activity of inducible nitric oxide synthase (iNOS). We demonstrate that the detrimental effect of the LPS on cNOS was manifested by the enzyme protein S nitrosylation, that was susceptible to inhibition by iNOS inhibitor, 1400 W. Further, we show that a peptide hormone, ghrelin, countered the LPS-induced changes in apoptosis and cNOS activity. This effect of ghrelin was reflected in the decrease in cNOS S-nitrosylation and the increase in phosphorylation. Our findings imply that P. gingivalis-induced disturbances in the acinar cell NO signaling pathways result from upregulation in iNOS-derived NO that causes cNOS S nitrosylation that interferes with its activation through phosphorylation. We also show that ghrelin protection against P. gingivalis-induced disturbances involves cNOS activation associated with a decrease in its S-nitrosylation and the increase in phosphorylation. PMID- 21637355 TI - Biomarkers of radiosensitivity in a-bomb survivors pregnant at the time of bombings in hiroshima and nagasaki. AB - Purpose. There is evidence in the literature of increased maternal radiosensitivity during pregnancy. Materials and Methods. We tested this hypothesis using information from the atomic-bomb survivor cohort, that is, the Adult Health Study database at the Radiation Effects Research Foundation, which contains data from a cohort of women who were pregnant at the time of the bombings of Hiroshima and Nagasaki. Previous evaluation has demonstrated long term radiation dose-response effects. Results/Conclusions. Data on approximately 250 women were available to assess dose-response rates for serum cholesterol, white blood cell count, erythrocyte sedimentation rate, and serum hemoglobin, and on approximately 85 women for stable chromosome aberrations, glycophorin A locus mutations, and naive CD4 T-cell counts. Although there is no statistically significant evidence of increased radiosensitivity in pregnant women, the increased slope of the linear trend line in the third trimester with respect to stable chromosome aberrations is suggestive of an increased radiosensitivity. PMID- 21637356 TI - Domestic Violence during Pregnancy and Mental Health: Exploratory Study in Primary Health Centers in Penalolen. AB - Objective. To determine the prevalence of domestic violence in a sample of pregnant women attending Primary Health Centers in Penalolen (Penalolen is a low income district in the Metropolitan Region in Santiago de Chile.), to explore risk facts for domestic violence during pregnancy, and to establish associations with their psychological health. Method and Materials. 256 pregnant women were assessed with a domestic violence screening and a questionnaire on mental symptoms. Frequency and correlations analysis were developed. Results. 5, 9% of the participants reported physical violence during current pregnancy. Emotional violence ascended to 30, 1% of the cases. Main risk facts found were as follows: having suffered violence along lifetime and physical violence during the last year. Anxiety and depressive symptoms positively correlated to domestic violence during pregnancy, but also to previous domestic violence experiences. Conclusions. Domestic violence during pregnancy is a prevalent problem and domestic violence history constitutes an alert to its occurrence. Positive and significant association to psychological disturbances suggests the need to detect it early during antenatal care. PMID- 21637357 TI - Inguinal endometriosis: an uncommon differential diagnosis as an inguinal tumour. AB - Inguinal endometriosis can present as a rare tumor when it occurs outside the abdomen and pelvis. We present a patient with a painful tumor in the right inguinal region, where its diagnosis was made before the operation. This enabled a conservative excisional surgery to be performed. The literature is reviewed and the diagnosis as well as management are discussed. PMID- 21637358 TI - Management of Precancerous Lesions of the Uterine Cervix according to Demographic Data. AB - Aims. Worldwide, cervical cancer is the fifth most deadly cancer in women, but screening prevents cancer by detecting precancerous lesions. The purpose of this study is to present the treatment profile for precancerous lesions of the uterine cervix, according to demographic data. Methods. An annual retrospective study was conducted in two public primary health care centres in Greece. The total number of Pap smears and colposcopies performed as well as the management of women with cervical intraepithelial neoplasia was collected and analysed. Results. Demographic characteristics and correlations with levels of Cervical Intraepithelial Neoplasia (CIN) and treatment path are presented. For each case, we noted the patients' age, the marital and educational status, and the professional and insurance type. From a total of 238 diagnostic procedures, 118 (49.5%) showed precancerous lesions, 83.3% of these were high grade while 16.7% were low grade. Conclusions. This study provides an estimate of the extensiveness of precancerous lesions of the uterine cervix. Management of CIN should be accounted for when balancing the benefits and unfavourable effects of this screening. PMID- 21637359 TI - An Evaluation of the Timing and Use of Healthcare during Pregnancy in Birmingham, UK and Pretoria, South Africa. AB - Objective. A pilot study to compare the rates of antenatal healthcare use in Birmingham, UK and Pretoria, South Africa, and identify differences in knowledge and perception of antenatal healthcare. Subjects. 62 women, 31 at each location <24 hours after delivery. Results. Women from Birmingham use healthcare services earlier (P <= .0001) and more often during pregnancy (P <= .0001). Women from Birmingham identified more conditions that may affect pregnancy (median 6 versus 3 reasons) and were less aware of HIV. In addition they perceived antenatal healthcare as relatively more important for advice and reassurance about pregnancy, whilst women from Pretoria had more problems with transport and clinic overcrowding. Conclusions. Increasing education on the importance of antenatal healthcare and medical problems during pregnancy may help improve antenatal healthcare use in Pretoria. Improving transport links and overcrowding in clinics in Pretoria may also help increase use. Measuring maternal outcomes and confirming these findings in a larger population are important for future studies. PMID- 21637360 TI - Breast metastatic localization of signet-ring cell gastric carcinoma. AB - Metastatic tumors in the breast are quite rare and constitute 0,5 to 6% of all breast malignancies. They often occur in a polymetastatic context. Gastrointestinal lesions rarely metastasize to the breast. The first case of a metastasis deposit to the breast and ovary from gastric signet-ring cell carcinoma was reported in the literature in 1999. Since this report, only 5 cases have been reported. We present a case report of a 37-year-old woman who complained of a lump in the left breast. Two months earlier, the woman underwent a subtotal gastrectomy and a total hysterectomy with double anexectomy, which histologically was diagnosed of gastric signet-ring carcinoma, disseminated with Krukenberg's tumor. In those days, the patient was following a chemotherapy treatment. A core needle biopsy of the lesion in left breast revealed cells with signet-ring features, with probably gastric origin. PMID- 21637361 TI - Accuracy of prenatal diagnosis in elective termination of pregnancy: 385 cases from 2000 to 2007. AB - Objective. To evaluate the quality of prenatal results in all cases of termination of pregnancy (TOP) due to fetal abnormalities in a tertiary prenatal diagnosis center. Material and Methods. Retrospective analysis of the 385 TOP performed on our department due to fetal abnormalities between January 1, 2000, and December 31, 2007. We compared all data for agreement between the ultrasound, genetic, and postmortem findings, regarding the abnormalities identified in the etiological diagnosis and its prognosis. Results. Chromosome abnormalities were the most common indication for TOP (39%), followed by abnormalities of CNS (20%), monogenic disorders (11%), sequences (9.6%), polimalformative syndromes (5.2%), and isolated congenital heart diseases (4%). Total agreement was 21%. Further abnormalities were identified in 79%. The data collected after TOP changed the etiologic diagnosis in 21% but the prognosis was changed in only one fetus. Discussion. This study corroborates the necessity of a multidisciplinary team in prenatal diagnosis centers. Their work remarkably improves the genetic counseling and represents an important aspect in quality control of the information given to a couple previously to a TOP. PMID- 21637362 TI - Spontaneous rupture of the imperforate hymen in an adolescent girl with hematocolpometra. AB - Background. Imperforate hymen is usually treated with hymenotomy, and the management after its spontaneous rupture is not very well known. Case. In this paper, we present spontaneous rupture of the imperforate hymen in a 13-year-old adolescent girl with hematocolpometra just before a planned hymenotomy operation. The patient was managed conservatively with a satisfactory outcome. Conclusion. Hymenotomy may not be needed in cases with spontaneous rupture of the imperforate hymen if adequate opening for menstrual discharge is warranted. PMID- 21637363 TI - Bilateral ovarian krukenberg tumor in a full-term pregnancy. AB - Objective. Krukenberg tumors in pregnancy are very rare and their management can present a dilemma for the obstetrician gynecologist. Case Report. We present the case of a G3P2002 who presented to us and the 38 weeks gestation with bilateral massive Krukenberg tumors. Despite at surgery and chemotherapy she died 3 months postpartum. Conclusion. Early detection followed by surgery and chemotherapy could possibly result in a favorable outcome with such patients. PMID- 21637364 TI - Low-Protein Diet during Lactation and Maternal Metabolism in Rats. AB - Some metabolic alterations were evaluated in Wistar rats which received control or low-protein (17%; 6%) diets, from the pregnancy until the end of lactation: control non-lactating (CNL), lactating (CL), low-protein non-lactating (LPNL) and lactating (LPL) groups. Despite the increased food intake by LPL dams, both LP groups reduced protein intake and final body mass was lower in LPL. Higher serum glucose occurred in both LP groups. Lactation induced lower insulin and glucagon levels, but these were reduced by LP diet. Prolactin levels rose in lactating, but were impaired in LPL, followed by losses of mammary gland (MAG) mass and, a fall in serum leptin in lactating dams. Lipid content also reduced in MAG and gonadal white adipose tissue of lactating and, in LPL, contributed to a decreased daily milk production, and consequent impairment of body mass gain by LPL pups. Liver mass, lipid content and ATP-citrate enzyme activity were increased by lactation, but malic enzyme and lipid: glycogen ratio elevated only in LPL. Conclusion. LP diet reduced the development of MAG and prolactin secretion which compromised milk production and pups growth. Moreover, this diet enhanced the store of lipid to glycogen ratio and suggests a higher risk of fatty liver development. PMID- 21637365 TI - Body mass index impacts in vitro fertilization stimulation. AB - The objective of the study was to prospectively determine if body mass index (BMI) is predictive of live birth rates in patients undergoing IVF. The prospective study enrolled 117 infertility patients with the primary outcome measure being IVF success rates. Mean BMI did not differ between patients with successful outcomes and those without successful outcomes. There was a significant positive correlation between BMI and the number of stimulated follicles (r = 0.19, P < .05). A significant negative correlation between BMI and ampules of gonadotropins used (r = -0.25, P < .01) and between BMI and days of stimulation (r = -0.19, P < .05) was noted. These data demonstrate that women with an elevated BMI produce more follicles, stimulate quicker, and require less gonadotropins during IVF. However, BMI did not have a significant effect on pregnancy outcome rates. PMID- 21637366 TI - Relative Roles of TGF-beta and IGFBP-5 in Idiopathic Pulmonary Fibrosis. AB - Although most evident in the skin, the process of scarring, or fibrosis, occurs in all major organs because of impaired epithelial self-renewal. No current therapy exists for Idiopathic pulmonary fibrosis. The major profibrotic factor is TGF-beta1 and developing inhibitors is an area of active research. Recently, IGFBP-5 has also been identified as a profibrotic factor, and studies suggest that, while both TGF-beta1 and IGFBP-5 activate mesenchymal cells to increase collagen and fibronectin production, their effects on epithelial cells are distinct. TGF-beta1 induces cell death and/or EMT in the epithelial cells, exacerbating the disruption of tissue architecture. In contrast, IGFBP-5 induces epithelial cell spreading over collagen or fibronectin matrices, increases secretion of laminin, the epithelial basement membrane, and enhances the survival of epithelial cells in nutrient-poor conditions, as exists in scar tissue. Thus, IGFBP-5 may enhance repair and may be an important target for antifibrotic therapies. PMID- 21637367 TI - Diffuse alveolar damage: a common phenomenon in progressive interstitial lung disorders. AB - It has become obvious that several interstitial lung diseases, and even viral lung infections, can progress rapidly, and exhibit similar features in their lung morphology. The final histopathological feature, common in these lung disorders, is diffuse alveolar damage (DAD). The histopathology of DAD is considered to represent end stage phenomenon in acutely behaving interstitial pneumonias, such as acute interstitial pneumonia (AIP) and acute exacerbations of idiopathic pulmonary fibrosis (IPF). Acute worsening and DAD may occur also in patients with nonspecific interstitial pneumonias (NSIPs), and even in severe viral lung infections where there is DAD histopathology in the lung. A better understanding of the mechanisms underlying the DAD reaction is needed to clarify the treatment for these serious lung diseases. There is an urgent need for international efforts for studying DAD-associated lung diseases, since the prognosis of these patients has been and is still dismal. PMID- 21637368 TI - Macrophage-derived biomarkers of idiopathic pulmonary fibrosis. AB - Idiopathic pulmonary fibrosis (IPF) is a severe, rapidly progressive diffuse lung disease. Several pathogenetic mechanisms have been hypothesized on the basis of the fibrotic lung damage occurring in this disease, and a potential profibrotic role of activated alveolar macrophages and their mediators in the pathogenesis of IPF was recently documented. This paper focuses on recent literature on potential biomarkers of IPF derived from activated alveolar macrophages. Biomarker discovery and clinical application are a recent topic of interest in the field of interstitial lung diseases (ILDs). Cytokines, CC-chemokines, and other macrophage produced mediators are the most promising prognostic biomarkers. Many molecules have been proposed in the literature as potential biomarker of IPF; however, a rigorous validation is needed to confirm their clinical utility. PMID- 21637369 TI - Subjects with Discordant Airways Obstruction: Lost between Spirometric Definitions of COPD. AB - Background. Since the FEV1/FVC ratio declines with age, using the fixed ratio of 0.70 leads to overdiagnosis of COPD in older populations and underdiagnosis among young adults. Objective. To evaluate whether discordant obstructive cases (FEV1/FVC < 0.70 but >=LLN) are a healthy population or have clinical features that would place them at increased risk. Methods. We used post-bronchodilator spirometry data from the population-based Austrian Burden of Obstructive Lung Disease (BOLD) study. Those with post-bronchodilator FEV1/FVC ratio =LLN but <0.70 were defined as discordant obstructive cases. Results. Discordant obstructive cases were more likely to be older, male and never-smokers. Additionally they had less respiratory symptoms and less severe impairment of FEV1. However, discordant obstructive cases reported significantly more often a diagnosis of heart disease than subjects with normal lung function (27.2% vs 7.3%, P = .015). Conclusion. The clinical profile of discordant obstructive cases includes potentially important comorbid disease. PMID- 21637370 TI - A Normal Range of KL-6/MUC1 Independent of Elevated SP-D Indicates a Better Prognosis in the Patients with Honeycombing on High-Resolution Computed Tomography. AB - Both SP-D and KL-6/MUC1 are established biomarkers of the interstitial pneumonias, including idiopathic pulmonary fibrosis (IPF), but the causes and clinical outcomes based on their independent effects are not known. Eleven asymptomatic patients, detected with honeycombing on high-resolution computed tomography (HRCT), were compared with 17 other IPF outpatients having slight respiratory symptoms and honeycombing as well. Although SP-D was increased in both groups, KL-6 was significantly higher in the symptomatic IPF group. When the patients (n = 11) having both biomarkers elevated were compared with the other patients (n = 6) with only SP-D elevated, the distribution of fibrotic lesions with honeycombing on HRCT was larger and the survival time was shorter in the patients having both biomarkers elevated. Immunohistochemical analysis also differentiated these biomarkers in the lung. These results suggest both a cause and the prognostic value of dissociation of these biomarkers. PMID- 21637371 TI - Amiloride-sensitive sodium channels and pulmonary edema. AB - The development of pulmonary edema can be considered as a combination of alveolar flooding via increased fluid filtration, impaired alveolar-capillary barrier integrity, and disturbed resolution due to decreased alveolar fluid clearance. An important mechanism regulating alveolar fluid clearance is sodium transport across the alveolar epithelium. Transepithelial sodium transport is largely dependent on the activity of sodium channels in alveolar epithelial cells. This paper describes how sodium channels contribute to alveolar fluid clearance under physiological conditions and how deregulation of sodium channel activity might contribute to the pathogenesis of lung diseases associated with pulmonary edema. Furthermore, sodium channels as putative molecular targets for the treatment of pulmonary edema are discussed. PMID- 21637372 TI - Gradual increase of high mobility group protein b1 in the lungs after the onset of acute exacerbation of idiopathic pulmonary fibrosis. AB - The pathogenesis of acute exacerbation of idiopathic pulmonary fibrosis (IPF) remains to be elucidated. To evaluate the roles of inflammatory mediators in acute exacerbation, the concentrations of high mobility group protein B1 (HMGB1), a chief mediator of acute lung injury, and 18 inflammatory cytokines were measured in the bronchoalveolar lavage fluid, serially sampled from seven IPF patients after the onset of acute exacerbation. HMGB1 gradually increased in the alveolar fluid after the onset of acute exacerbation, in positive correlation with monocytes chemotactic protein-1 (MCP-1), a potent fibrogenic mediator. In the lung tissues of eight IPF patients autopsied after acute exacerbation, intense cytoplasmic staining for HMGB1 was observed in the alveolar epithelial cells in alveolar capillary augmented lesions, where the capillary endothelial cells remarkably reduced the expression of thrombomodulin, an intrinsic antagonist of HMGB1. These results suggest pathogenic roles for HMGB1 and MCP-1 in the late phase of acute exacerbation of IPF. PMID- 21637373 TI - Postoperative Acute Exacerbation of IPF after Lung Resection for Primary Lung Cancer. AB - Idiopathic pulmonary fibrosis (IPF) is characterized by slowly progressive respiratory dysfunction. Nevertheless, some IPF patients experience acute exacerbations generally characterized by suddenly worsening and fatal respiratory failure with new lung opacities and pathological lesions of diffuse alveolar damage. Acute exacerbation of idiopathic pulmonary fibrosis (AEIPF) is a fatal disorder defined by rapid deterioration of IPF. The condition sometimes occurs in patients who underwent lung resection for primary lung cancer in the acute and subacute postoperative phases. The exact etiology and pathogenesis remain unknown, but the condition is characterized by diffuse alveolar damage superimposed on a background of IPF that probably occurs as a result of a massive lung injury due to some unknown factors. This systematic review shows that the outcome, however, is poor, with postoperative mortality ranging from 33.3% to 100%. In this paper, the etiology, risk factors, pathogenesis, therapy, prognosis, and predictors of postoperative AEIPF are described. PMID- 21637374 TI - Mos in the oocyte: how to use MAPK independently of growth factors and transcription to control meiotic divisions. AB - In many cell types, the mitogen-activated protein kinase (MAPK) also named extracellular signal-regulated kinase (ERK) is activated in response to a variety of extracellular growth factor-receptor interactions and leads to the transcriptional activation of immediate early genes, hereby influencing a number of tissue-specific biological activities, as cell proliferation, survival and differentiation. In one specific cell type however, the female germ cell, MAPK does not follow this canonical scheme. In oocytes, MAPK is activated independently of growth factors and tyrosine kinase receptors, acts independently of transcriptional regulation, plays a crucial role in controlling meiotic divisions, and is under the control of a peculiar upstream regulator, the kinase Mos. Mos was originally identified as the transforming gene of Moloney murine sarcoma virus and its cellular homologue was the first proto-oncogene to be molecularly cloned. What could be the specific roles of Mos that render it necessary for meiosis? Which unique functions could explain the evolutionary cost to have selected one gene to only serve for few hours in one very specific cell type? This review discusses the original features of MAPK activation by Mos and the roles of this module in oocytes. PMID- 21637375 TI - Integrin signaling in oligodendrocytes and its importance in CNS myelination. AB - Multiple sclerosis is characterized by repeated demyelinating attacks of the central nervous system (CNS) white matter tracts. To tailor novel therapeutics to halt or reverse disease process, we require a better understanding of oligodendrocyte biology and of the molecular mechanisms that initiate myelination. Cell extrinsic mechanisms regulate CNS myelination through the interaction of extracellular matrix proteins and their transmembrane receptors. The engagement of one such receptor family, the integrins, initiates intracellular signaling cascades that lead to changes in cell phenotype. Oligodendrocytes express a diverse array of integrins, and the expression of these receptors is developmentally regulated. Integrin-mediated signaling is crucial to the proliferation, survival, and maturation of oligodendrocytes through the activation of downstream signaling pathways involved in cytoskeletal remodeling. Here, we review the current understanding of this important signaling axis and its role in oligodendrocyte biology and ultimately in the myelination of axons within the CNS. PMID- 21637376 TI - Activation of mitogen-activated protein kinase in descending pain modulatory system. AB - The descending pain modulatory system is thought to undergo plastic changes following peripheral tissue injury and exerts bidirectional (facilitatory and inhibitory) influence on spinal nociceptive transmission. The mitogen-activated protein kinases (MAPKs) superfamily consists of four main members: the extracellular signal-regulated protein kinase1/2 (ERK1/2), the c-Jun N-terminal kinases (JNKs), the p38 MAPKs, and the ERK5. MAPKs not only regulate cell proliferation and survival but also play important roles in synaptic plasticity and memory formation. Recently, many studies have demonstrated that noxious stimuli activate MAPKs in several brain regions that are components of descending pain modulatory system. They are involved in pain perception and pain-related emotional responses. In addition, psychophysical stress also activates MAPKs in these brain structures. Greater appreciation of the convergence of mechanisms between noxious stimuli- and psychological stress-induced neuroplasticity is likely to lead to the identification of novel targets for a variety of pain syndromes. PMID- 21637377 TI - Integrins are the necessary links to hypertrophic growth in cardiomyocytes. AB - To compensate for hemodynamic overload of the heart, an event which stretches the myocardium, growth and survival signaling are activated in cardiac muscle cells (cardiomyocytes). Integrins serve as the signaling receptors of cardiomyocytes responsible for mechanotransduction toward intracellular signaling. The main integrin heterodimers on the cardiomyocyte surface are alpha(5)beta(1) and alpha(v)beta(3), and elimination of either beta(1) or beta(3) integrins impedes pressure-induced hypertrophic signaling and leads to increased mortality. The growth signaling pathways downstream of beta(1) and beta(3) integrins are well characterized. However, new integrin pathways responsible for inhibiting apoptosis induced by hemodynamic overload are emerging. beta(1) and beta(3) integrins activate differential survival signaling, yet both integrins initiate survival signaling downstream of ubiquitination and the kinase pathway including phosphoinositol-3-kinase (PI3K)/Akt. Further characterization of these integrin signaling mechanisms may lead to drug targets to prevent decompensation to heart failure. PMID- 21637378 TI - Regulation of ack-family nonreceptor tyrosine kinases. AB - Ack family non-receptor tyrosine kinases are unique with regard to their domain composition and regulatory properties. Human Ack1 (activated Cdc42-associated kinase) is ubiquitously expressed and is activated by signals that include growth factors and integrin-mediated cell adhesion. Stimulation leads to Ack1 autophosphorylation and to phosphorylation of additional residues in the C terminus. The N-terminal SAM domain is required for full activation. Ack1 exerts some of its effects via protein-protein interactions that are independent of its kinase activity. In the basal state, Ack1 activity is suppressed by an intramolecular interaction between the catalytic domain and the C-terminal region. Inappropriate Ack1 activation and signaling has been implicated in the development, progression, and metastasis of several forms of cancer. Thus, there is increasing interest in Ack1 as a drug target, and studies of the regulatory properties of the enzyme may reveal features that can be exploited in inhibitor design. PMID- 21637379 TI - Mitogen-Activated Protein Kinases and Reactive Oxygen Species: How Can ROS Activate MAPK Pathways? AB - Mitogen-activated protein kinases (MAPKs) are serine-threonine protein kinases that play the major role in signal transduction from the cell surface to the nucleus. MAPKs, which consist of growth factor-regulated extracellular signal related kinases (ERKs), and the stress-activated MAPKs, c-jun NH(2)-terminal kinases (JNKs) and p38 MAPKs, are part of a three-kinase signaling module composed of the MAPK, an MAPK kinase (MAP2K) and an MAPK kinase (MAP3K). MAP3Ks phosphorylate MAP2Ks, which in turn activate MAPKs. MAPK phosphatases (MKPs), which recognize the TXY amino acid motif present in MAPKs, dephosphorylate and deactivate MAPKs. MAPK pathways are known to be influenced not only by receptor ligand interactions, but also by different stressors placed on the cell. One type of stress that induces potential activation of MAPK pathways is the oxidative stress caused by reactive oxygen species (ROS). Generally, increased ROS production in a cell leads to the activation of ERKs, JNKs, or p38 MAPKs, but the mechanisms by which ROS can activate these kinases are unclear. Oxidative modifications of MAPK signaling proteins and inactivation and/or degradation of MKPs may provide the plausible mechanisms for activation of MAPK pathways by ROS, which will be reviewed in this paper. PMID- 21637380 TI - The Functional Crosstalk between HER2 Tyrosine Kinase and TGF-beta Signaling in Breast Cancer Malignancy. AB - Accumulating evidence indicates a functional crosstalk between the HER2 (ErbB2) tyrosine kinase and the TGF-beta signaling mediated by its serine/threonine kinase receptors. In HER2-overexpressing breast cancer, this crosstalk results in increased cancer cell proliferation, survival and invasion, accelerated cancer progression and metastasis in animal models, and resistance to chemotherapy and HER2-targeted therapy. The transformed cellular context with constitutively active HER2 signaling, as a consequence of HER2 gene amplification or overexpression, converts TGF-beta from a tumor suppressor to a malignancy promoting factor. TGF-beta, in turn, potentiates oncogenic HER2 signaling by inducing shedding of the ErbB ligands and clustering of HER2 with integrins. In addition, TGF-beta is associated with resistance to trastuzumab, an anti-HER2 therapeutic antibody. Recent mechanistic studies indicate that TGF-beta and HER2 cooperate through both Smad-dependent and independent mechanisms. Blockade of HER2:TGF-beta crosstalk may significantly enhance the efficiency of conventional therapies in breast cancer patients with HER2 overexpression. PMID- 21637381 TI - The role of specific mitogen-activated protein kinase signaling cascades in the regulation of steroidogenesis. AB - Mitogen-activated protein kinases (MAPKs) comprise a family of serine/threonine kinases that are activated by a large variety of extracellular stimuli and play integral roles in controlling many cellular processes, from the cell surface to the nucleus. The MAPK family includes four distinct MAPK cascades, that is, extracellular signal-regulated kinase 1/2 (ERK1/2), p38 MAPK, c-Jun N-terminal kinase or stress-activated protein kinase, and ERK5. These MAPKs are essentially operated through three-tiered consecutive phosphorylation events catalyzed by a MAPK kinase kinase, a MAPK kinase, and a MAPK. MAPKs lie in protein kinase cascades. The MAPK signaling pathways have been demonstrated to be associated with events regulating the expression of the steroidogenic acute regulatory protein (StAR) and steroidogenesis in steroidogenic tissues. However, it has become clear that the regulation of MAPK-dependent StAR expression and steroid synthesis is a complex process and is context dependent. This paper summarizes the current level of understanding concerning the roles of the MAPK signaling cascades in the regulation of StAR expression and steroidogenesis in different steroidogenic cell models. PMID- 21637382 TI - Raf-1 activation prevents caspase 9 processing downstream of apoptosome formation. AB - In many cell types, growth factor removal induces the release of cytochrome-c from mitochondria that leads to activation of caspase-9 in the apoptosome complex. Here, we show that sustained stimulation of the Raf-1/MAPK1,3 pathway prevents caspase-9 activation induced by serum depletion in CCL39/DeltaRaf-1:ER fibroblasts. The protective effect mediated by Raf-1 is sensitive to MEK inhibition that is sufficient to induce caspase-9 cleavage in exponentially growing cells. Raf-1 activation does not inhibit the release of cytochrome-c from mitochondria while preventing caspase-9 activation. Gel filtration chromatography analysis of apoptosome formation in cells shows that Raf-1/MAPK1,3 activation does not interfere with APAF-1 oligomerization and recruitment of caspase 9. Raf 1-mediated caspase-9 inhibition is sensitive to emetine, indicating that the protective mechanism requires protein synthesis. However, the Raf/MAPK1,3 pathway does not regulate XIAP. Taken together, these results indicate that the Raf 1/MAPK1,3 pathway controls an apoptosis regulator that prevents caspase-9 activation in the apoptosome complex. PMID- 21637383 TI - Receptor tyrosine kinases in kidney development. AB - The kidney plays a fundamental role in the regulation of arterial blood pressure and fluid/electrolyte homeostasis. As congenital anomalies of the kidney and urinary tract (CAKUT) constitute one of the most common human birth defects, improved understanding of the cellular and molecular mechanisms that lead to CAKUT is critical. Accumulating evidence indicates that aberrant signaling via receptor tyrosine kinases (RTKs) is causally linked to CAKUT. Upon activation by their ligands, RTKs dimerize, undergo autophosphorylation on specific tyrosine residues, and interact with adaptor proteins to activate intracellular signal transduction pathways that regulate diverse cell behaviours such as cell proliferation, survival, and movement. Here, we review the current understanding of role of RTKs and their downstream signaling pathways in the pathogenesis of CAKUT. PMID- 21637384 TI - The Dynamical Systems Properties of the HOG Signaling Cascade. AB - The High Osmolarity Glycerol (HOG) MAP kinase pathway in the budding yeast Saccharomyces cerevisiae is one of the best characterized model signaling pathways. The pathway processes external signals of increased osmolarity into appropriate physiological responses within the yeast cell. Recent advances in microfluidic technology coupled with quantitative modeling, and techniques from reverse systems engineering have allowed yet further insight into this already well-understood pathway. These new techniques are essential for understanding the dynamical processes at play when cells process external stimuli into biological responses. They are widely applicable to other signaling pathways of interest. Here, we review the recent advances brought by these approaches in the context of understanding the dynamics of the HOG pathway signaling. PMID- 21637385 TI - Parasite mitogen-activated protein kinases as drug discovery targets to treat human protozoan pathogens. AB - Protozoan pathogens are a highly diverse group of unicellular organisms, several of which are significant human pathogens. One group of protozoan pathogens includes obligate intracellular parasites such as agents of malaria, leishmaniasis, babesiosis, and toxoplasmosis. The other group includes extracellular pathogens such as agents of giardiasis and amebiasis. An unfortunate unifying theme for most human protozoan pathogens is that highly effective treatments for them are generally lacking. We will review targeting protozoan mitogen-activated protein kinases (MAPKs) as a novel drug discovery approach towards developing better therapies, focusing on Plasmodia, Leishmania, and Toxoplasma, about which the most is known. PMID- 21637386 TI - Role of innate immunity in diabetes and metabolism: recent progress in the study of inflammasomes. AB - Type 1 diabetes is one of the classical examples of organ- specific autoimmune diseases characterized by lymphocytic infiltration or inflammation in pancreatic islets called 'insulitis'. In contrast, type 2 diabetes has been traditionally regarded as a metabolic disorder with a pathogenesis that is totally different from that of type 1 diabetes. However, recent investigation has revealed contribution of chronic inflammation in the pathogenesis of type 2 diabetes. In addition to type 2 diabetes, the role of chronic inflammation is being appreciated in a wide variety of metabolic disorders such as obesity, metabolic syndrome, and atherosclerosis. In this review, we will cover the role of innate immunity in the pathogenesis of metabolic disorders with an emphasis on NLRP3. PMID- 21637387 TI - Characterization of CTL Clones Specific for Single Antigen, H60 Minor Histocompatibility Antigen. AB - BACKGROUND: Disparities of Minor H antigens can induce graft rejection after MHC matched transplantation. H60 has been characterized as a dominant antigen expressed on hematopoietic cells and considered to be an ideal model antigen for study on graft-versus-leukemia effect. METHODS: Splenocytes from C57BL/6 mice immunized with H60 congenic splenocytes were used for establishment of H60 specific CTL clones. Then the clones were characterized for proliferation capacity and cytotoxicity after stimulation with H60. Clone #14, #15, and #23 were tested for the TCR binding avidity to H60-peptide/H-2K(b) and analyzed for TCR sequences. RESULTS: H60-specific CTL clones showed different levels of proliferation capacity and cytotoxic activity to H60-stimulation. Clones #14, #15, and #23 showed high proliferation activity, high cytotoxicity, and low activities on both aspects, respectively, and have TCRs with different binding avidities to H60-peptide/H-2K(b) with t(1/2) values of 4.87, 6.92, and 13.03 minutes, respectively. The TCR usages were Valpha12D-3-01+Jalpha11-01 and Vbeta12 1-01+Dbeta1-01+J2-7-01 for clone #14, Valpha13D-1-02+Jalpha34-02 and Vbeta13-1 02+Dbeta2-01+Jbeta2-7-01 for clone #15, and Valpha16D+Jalpha45-01 and Vbeta12-1 01+Dbeta1-01+Jbeta2-5-01 for clone #23. CONCLUSION: The results will be useful for modeling GVL and generation TCR transgenic mouse. PMID- 21637388 TI - Dietary Aloe Reduces Adipogenesis via the Activation of AMPK and Suppresses Obesity-related Inflammation in Obese Mice. AB - BACKGROUND: Metabolic disorders, including type II diabetes and obesity, present major health risks in industrialized countries. AMP-activated protein kinase (AMPK) has become the focus of a great deal of attention as a novel therapeutic target for the treatment of metabolic syndromes. In this study, we evaluated whether dietary aloe could reduce obesity-induced inflammation and adipogenesis. METHODS: Male C57BL/6 obese mice fed a high-fat diet for 54 days received a supplement of aloe formula (PAG, ALS, Aloe QDM, and Aloe QDM complex) or pioglitazone (PGZ) and were compared with unsupplemented controls (high-fat diet; HFD) or mice fed a regular diet (RD). RT-PCR and western blot analysis were used to quantify the expression of obesity-induced inflammation. RESULTS: Aloe QDM complex down-regulated fat size through suppressed expression of scavenger receptors on adipose tissue macrophages (ATMs) compared with HFD. Both white adipose tissue (WATs) and muscle exhibited increased AMPK activation through aloe supplementation, and in particular, the Aloe QDM complex. Obesity-induced inflammatory cytokines (IL-1beta and -6) and HIF1alpha mRNA and protein were decreased markedly, as was macrophage infiltration by the Aloe QDM complex. Further, the Aloe QDM complex decreased the translocation of NF-kappaB p65 from the cytosol in the WAT. CONCLUSION: Dietary aloe formula reduced obesity-induced inflammatory responses by activation of AMPK in muscle and suppression of proinflammatory cytokines in the WAT. Additionally, the expression of scavenger receptors in the ATM and activation of AMPK in WAT led to reduction in the percent of body fat. Thus, we suggest that the effect of the Aloe QDM complex in the WAT and muscle are related to activation of AMPK and its use as a nutritional intervention against T2D and obesity-related inflammation. PMID- 21637389 TI - Characterization of Monoclonal Antibodies against Human Leukocyte Common Antigen (CD45). AB - BACKGROUND: The leukocyte common antigen (CD45) is a transmembrane-type protein tyrosine phosphatase that has five isoforms. METHODS: We generated seven murine mAbs against human CD45 by injecting cells from different origins, such as human thymocytes, PBMCs, and leukemic cell lines. By using various immunological methods including flow cytometry, immunohistochemistry, and immunoprecipitation, we evaluated the reactivity of those mAbs to CD45 of thymus as well as tonsil lysates. Furthermore, we transiently transfected COS-7 cells with each of gene constructs that express five human CD45 isoforms respectively, and examined the specificities of the mAbs against the transfected isoforms. RESULTS: In case of thymocytes, lymphocytes, and monocytes, all the seven mAbs demonstrated positive reactivities whereas none was reactive to erythrocytes and platelets. The majority of immune cells in formalin-fixed paraffin-embedded thymus and tonsil tissues displayed strong membranous immunoreactivity, and the main antigen was detected near 220 kDa in all cases. Among the mAbs, four mAbs (AP4, DN11, SHL-1, and P6) recognized a region commonly present in all the five isoforms. One mAb, YG27, recognized four isoforms (ABC, AB, BC, and O). Two mAbs, P1 and P14, recognized the isoforms that contain exon A encoded regions (ABC and AB). CONCLUSION: In this study, we confirmed that AP4, DN11, SHL-1, YG27 and P6, are mAbs reactive with the CD45 antigen whereas P1 and P14 are reactive with the CD45RA antigen. PMID- 21637391 TI - Using Evidence to Guide but not Dictate Practice. PMID- 21637390 TI - Mycobacterial Heparin-binding Hemagglutinin Antigen Activates Inflammatory Responses through PI3-K/Akt, NF-kappaB, and MAPK Pathways. AB - BACKGROUND: Mycobacterium tuberculosis (Mtb) heparin binding hemagglutinin (HBHA) is an Ag known to evoke effective host immune responses during tuberculosis infection. However, the molecular basis of the host immune response to HBHA has not been fully characterized. In this study, we examined the molecular mechanisms by which HBHA can induce the expression of proinflammatory cytokines in macrophages. METHODS: HBHA-induced mRNA and protein levels of proinflammatory cytokines were determined in bone marrow-derived macrophages (BMDMs) using RT-PCR and ELISA analysis. The roles of intracellular signaling pathways for NF-kappaB, PI3-K/Akt, and MAPKs were investigated in macrophage proinflammatory responses after stimulation with HBHA. RESULTS: HBHA robustly activated the expression of mRNA and protein of both TNF-alpha and IL-6, and induced phosphorylation of NF kappaB, Akt, and MAPKs in BMDMs. Both TNF-alpha and IL-6 production by HBHA was regulated by the NF-kappaB, PI3-K, and MAPK pathways. Furthermore, PI3-K activity was required for the HBHA-induced activation of ERK1/2 and p38 MAPK, but not JNK, pathways. CONCLUSION: These data suggest that mycobacterial HBHA significantly induces proinflammatory responses through crosstalk between the PI3-K and MAPK pathways in macrophages. PMID- 21637392 TI - Blood pressure measurement guidelines for physical therapists. AB - Vital sign measurement and assessment are important components of the review of systems in a physical therapy examination for individuals with and without documented cardiopulmonary disease. The measurement of blood pressure gives the therapist information regarding the patient's baseline cardiovascular status, response to exercise/activity, and guides exercise prescription. Accurate measurement of blood pressure is critical for making appropriate clinical decisions especially if physical therapists wish to play an important role as primary health care providers. The purpose of this paper is to present recommended guidelines for blood pressure measurement by physical therapists and physical therapist assistants. PMID- 21637393 TI - Case report: exercise in a patient with acute decompensated heart failure receiving positive inotropic therapy. AB - BACKGROUND AND PURPOSE: The projected increase in persons with advanced heart failure and associated costs warrant the examination of exercise in patients receiving inotropic therapy. Literature supports the use of exercise and inotropic therapy in the treatment of patients with advanced heart failure. The purposes of this paper are to illustrate the use of exercise prescription and outcomes assessment with a 6-minute walk test in a patient with acute decompensated heart failure receiving tailored therapy with dobutamine and to discuss potential relationships resulting in observed improvements. CASE DESCRIPTION: A 67-year old man was admitted to an acute care hospital with acute decompensated heart failure for tailored medical therapy including dobutamine. The patient received 14 days of tailored medical therapy, of which 12 days included exercise training by a physical therapist. OUTCOMES: Functional outcomes showed a clinically significant improvement in distance walked and improvement in the cardiorespiratory response. The improvement in estimated peak oxygen consumption was 7% greater than that predicted to be from tailored medical therapy. DISCUSSION: Exercise was safely provided to a patient hospitalized with advanced heart failure on continuous inotropic therapy. The 6-minute walk test was effectively used to prescribe exercise and examine patient outcomes. PMID- 21637394 TI - THE LINDA CRANE MEMORIAL LECTUR: Leading Leaders: A Vision for Our Centennial Years. AB - At 90 years of age, the APTA may be facing some of the greatest national and global challenges of its history. Membership has grown from 238 in 1921 to over 70,000 in 2011, but the expansion of the APTA may be restrictive to individual participation. A leadership gap appears imminent in practice and education. Fostering every member to understand the APTA and its great work is essential to ensuring a profession that lives its core values and meets societal needs. The Linda Crane Memorial Lecture in 2011 celebrated a vision of the APTA's 100th birthday with every member serving as a "professional centenarian" who stewards the organization to continued greatness. PMID- 21637395 TI - Low typing endurance in keyboard workers with work-related upper limb disorder. AB - OBJECTIVE: To compare results of typing endurance and pain before and after a standardized functional test. DESIGN: A standardized previously published typing test on a standard QWERTY keyboard. SETTING: An outpatient hospital environment. PARTICIPANTS: Sixty-one keyboard and mouse operating patients with WRULD and six normal controls. MAIN OUTCOME MEASURE: Pain severity before and after the test, typing endurance and speed were recorded. RESULTS: Thirty-two patients could not complete the test before pain reached VAS 5 and this group only typed a mean of 11 minutes. The control group and the remaining group of 29 patients completed the test. Two-tailed student T test was used for evaluation. The endurance was significantly shorter in the patient group that could not complete the test (P < 0.00001) and the pain levels were also higher in this group both before (P = 0.01) and after the test (P = 0.0003). Both patient groups had more pain in the right than the left hand, both before and after typing. CONCLUSIONS: Low typing endurance correlates statistically with more resting pain in keyboard and mouse operators with work-related upper limb disorder and statistically more pain after a standardized typing test. As the right hands had higher pain levels, typing alone may not be the cause of the pain as the left hand on a QWERTY keyboard does relative more keystrokes than the right hand. PMID- 21637396 TI - The incidence of peripheral arterial embolism in association with a patent foramen ovale (right-to-left shunt). AB - OBJECTIVES: The aim of this study was to examine a cohort of patients who had suffered an arterial embolism to see whether a patent foramen ovale (PFO) was an identifiable cause. DESIGN: This study was conducted in two parts; a retrospective limb involving an audit of patient records over a period of 10 years, and a prospective limb including selected patients from that audit to search for a PFO using an agitated saline test with transcranial Doppler ultrasound monitoring of the anterior cerebral artery. Data on patients with peripheral vascular disease were collected using a structured questionnaire. SETTING: A clinical vascular department. All patients were seen in the vascular outpatients clinic. PARTICIPANTS: Patients who had been identified from a retrospective search based on the headline diagnosis of arterial embolus. Collected data on the 71 patients revealed that 75% had predisposing factors for DVT, 70% were male smokers, and 84.4% had a significant past history of vascular symptoms. MAIN OUTCOME MEASURES: Whether or not patients identified as having a possible PFO actually had one on objective testing with transcranial Doppler assessment of the cerebral circulation with an agitated saline solution. RESULTS: Fifteen patients who were suspected of having a PFO were selected from these 71 patients; 12 of these were found to have no PFO on testing, and three had already undergone a percutaneous PFO closure. CONCLUSION: The incidence of a PFO in this small study group is no higher than that found in the general population (3/15, 20%). There was high prevalence of male smokers with associated predisposing factors leading to a DVT. PMID- 21637397 TI - Emergency first presentation of colorectal cancer following air travel: a case series. PMID- 21637398 TI - Patient information in otorhinolaryngology: a prospective audit. AB - OBJECTIVES: The aim of patient information is to involve patients in their condition and their treatment. The literature states that good information can improve medical outcomes, reduce patient anxiety and that patients want access to it. We wanted to calculate the provision of written patient information to ENT day-case patients, measure information recall and patient satisfaction. DESIGN: A prospective audit cycle. The first cycle of the audit studied patients receiving current practice, where verbal information was provided but written patient information was not routine. Following a departmental drive towards provision of written patient information, a second cycle was audited. A questionnaire on admission to the ward on the day of surgery was used to measure outcomes. SETTING: The ENT Department of a UK university teaching hospital. MAIN OUTCOME MEASURES: The number of patients receiving written patient information, the rate of recall of complications and patient satisfaction with the information provided. PARTICIPANTS: One hundred patients undergoing day-case surgery were included. The first cycle of the audit studied 50 consecutive patients, receiving current practice. The second cycle, following implementation of change, studied a further 50 consecutive patients. RESULTS: Following a departmental drive towards provision of patient information, 64% of patients received written patient information improving the rate of recall of the majority of complications from 24% to 52%. There was no significant difference in patient satisfaction between groups. CONCLUSIONS: Written patient information leaflets are a useful tool to improve recall of information given to patients, in order to facilitate informed consent. PMID- 21637399 TI - Liquefying panniculitis associated with intraductal papillary mucinous neoplasm. PMID- 21637400 TI - Career decision difficulties post foundation training - the medical student perspective. AB - OBJECTIVES: Since 2005, newly qualified doctors in the UK have had their time to prepare for career subspecialization application cut short to 16 months. To have enough time to become a competitive applicant, the choice of specialization may now have to be made as early as in medical school. This study aimed to assess how prepared medical students are towards committing to a specialty while in medical school, and their opinion about having to make such a decision. DESIGN: A cross sectional questionnaire. A list of all career specialties available to doctors at the point of specialization was provided and asked students to rank their top choice. An assessment of the certainty of their choice was then determined. SETTING: Questionnaires were distributed at the end of an optional final year medical student academic meeting held at a leading London medical school university. PARTICIPANTS: One hundred and thirty final year students attended the meeting. Questionnaires were distributed to all attenders; 115 responses were collected. MAIN OUTCOME MEASURES: The certainty of career specialization choice was assessed in qualitative form, with responses ranging from 'not likely', 'maybe', 'probably', 'almost certainly' and 'definitely'. Their feelings in having to decide upon career specialty while in medical school was assessed through either a 'yes' or 'no' response. RESULTS: A total of 115 responses were collected. The second most common selection was the 'undecided' option at 15%. The highest certainty factor occurred at 'maybe' with 41% and progressively fewer responses occurred as the certainty factor increased, with only 10% at 'definitely'; 95% voted 'no' to having to decide what they want to specialize in by the end of medical school. CONCLUSIONS: The majority of students have yet to commit to a specialty and almost all agree that they should not have to decide what they want to specialize in by the end of medical school. There is thus greater responsibility from medical schools to incorporate more career discussions into their syllabus. PMID- 21637401 TI - Clopidogrel-induced liver failure. PMID- 21637402 TI - Acute gastric dilatation causing respiratory distress. PMID- 21637403 TI - Chronic fatigue syndrome in the media: a content analysis of newspaper articles. AB - OBJECTIVES: Although cognitive behavioural therapy and graded exercise treatment are recognized evidence-based treatments for chronic fatigue syndrome/myalgic encephalomyelitis (ME), their use is still considered controversial by some patient groups. This debate has been reflected in the media, where many patients gather health information. The aim of this study was to examine how treatment for chronic fatigue syndrome/ME is described in the newspaper media. DESIGN: Content analysis of newspaper articles. SETTING: The digitalized media archive Atekst was used to identify Norwegian newspaper articles where chronic fatigue syndrome/ME was mentioned. PARTICIPANTS: Norwegian newspaper articles published over a 20 month period, from 1 January 2008 to 31 August 2009. MAIN OUTCOME MEASURES: Statements regarding efficiency of various types of treatment for chronic fatigue syndrome/ME and the related source of the treatment advice. Statements were categorized as being either positive or negative towards evidence-based or alternative treatment. RESULTS: One hundred and twenty-two statements regarding treatment of chronic fatigue syndrome/ME were identified among 123 newspaper articles. The most frequent statements were positive statements towards alternative treatment Lightning Process (26.2%), negative statements towards evidence-based treatments (22.1%), and positive statements towards other alternative treatment interventions (22.1%). Only 14.8% of the statements were positive towards evidence-based treatment. Case-subjects were the most frequently cited sources, accounting for 35.2% of the statements, followed by physicians and the Norwegian ME association. CONCLUSIONS: Statements regarding treatment for chronic fatigue syndrome/ME in newspapers are mainly pro-alternative treatment and against evidence-based treatment. The media has great potential to influence individual choices. The unbalanced reporting of treatment options for chronic fatigue syndrome/ME in the media is potentially harmful. PMID- 21637404 TI - Normal inflammatory markers in appendicitis: evidence from two independent cohort studies. AB - OBJECTIVES: Acute appendicitis is a common surgical condition which can lead to severe complications. Recent work suggested that patients experiencing right lower abdominal pain, with normal white cell count (WCC) and C-reactive protein (CRP) are unlikely to have acute appendicitis and can be discharged. We present two independent data-sets that suggest that this strategy may not be risk-free. DESIGN: Retrospective cohort study of consecutive patients from two district general hospitals. Sensitivity and specificity of CRP, WCC and neutrophil count (NC) in predicting appendicitis were calculated. Markers were analysed using Fisher's exact test and Kruskul-Wallace test. SETTING: Two district general hospitals in the UK. PARTICIPANTS: Patients undergoing appendicectomy for suspected appendicitis. MAIN OUTCOME MEASURES: Inflammatory markers and appendix histology. RESULTS: A total of 297 patients were included. Appendicitis occurred in four patients with normal CRP, WCC and NC in centre A and 13 patients in centre B. The sensitivity of all three markers combined was 94% (centre A) and 92% (centre B). The specificity was 60% (centre A) and 64% (centre B). No single marker could differentiate uncomplicated and complicated appendicitis, but a raised NC or a CRP >35.5 mg/l predicted complicated appendicitis. CRP, WCC and NC combined differentiated between patients with a normal appendix, uncomplicated appendicitis and complicated appendicitis. CONCLUSIONS: Appendicitis in the presence of normal inflammatory markers is not uncommon. We disagree with the view of Sengupta et al. who suggest that patients with normal WCC and CRP are unlikely to have appendicitis, and recommend that clinicians be wary of normal inflammatory markers in patients with a high clinical suspicion of appendicitis. PMID- 21637405 TI - beta-globin haplotypes in normal and hemoglobinopathic individuals from Reconcavo Baiano, State of Bahia, Brazil. AB - Five restriction site polymorphisms in the beta-globin gene cluster (HincII-5' epsilon, HindIII-(G) gamma, HindIII-(A) gamma, HincII- psibeta1 and HincII-3' psibeta1) were analyzed in three populations (n = 114) from Reconcavo Baiano, State of Bahia, Brazil. The groups included two urban populations from the towns of Cachoeira and Maragojipe and one rural Afro-descendant population, known as the "quilombo community", from Cachoeira municipality. The number of haplotypes found in the populations ranged from 10 to 13, which indicated higher diversity than in the parental populations. The haplotypes 2 (+ - - - -), 3 (- - - - +), 4 (- + - - +) and 6 (- + + - +) on the beta(A) chromosomes were the most common, and two haplotypes, 9 (- + + + +) and 14 (+ + - - +), were found exclusively in the Maragojipe population. The other haplotypes (1, 5, 9, 11, 12, 13, 14 and 16) had lower frequencies. Restriction site analysis and the derived haplotypes indicated homogeneity among the populations. Thirty-two individuals with hemoglobinopathies (17 sickle cell disease, 12 HbSC disease and 3 HbCC disease) were also analyzed. The haplotype frequencies of these patients differed significantly from those of the general population. In the sickle cell disease subgroup, the predominant haplotypes were BEN (Benin) and CAR (Central African Republic), with frequencies of 52.9% and 32.4%, respectively. The high frequency of the BEN haplotype agreed with the historical origin of the afro-descendant population in the state of Bahia. However, this frequency differed from that of Salvador, the state capital, where the CAR and BEN haplotypes have similar frequencies, probably as a consequence of domestic slave trade and subsequent internal migrations to other regions of Brazil. PMID- 21637406 TI - Micronuclei formation in liver fibrosis samples from patients infected by hepatitis C virus. AB - Genetic research on fibrosis outset and its progression in chronic hepatitis (CH) by hepatitis C virus (HCV) are limited. The lack of cytogenetic data led us to investigate the presence of micronuclei (MNi), as a sign of genomic damage. Hepatocytes of hepatic parenchyma from 62 cases diagnosed with CH associated with HCV and displaying different degrees of fibrosis (F1-F4) were analyzed. These data were compared to 15 cases without fibrosis (F0). Twelve healthy liver parenchyma samples were included as control. All samples were obtained from paraffin-embedded archival material. Micronucleated hepatocytes (MN-Heps) were analyzed through Feulgen/Fast-green staining. Results showed that the rates of MN Heps in the F4 group were statistically significant (p < 0.05) and higher than those in the control group. Like results were also obtained on comparing F4 with F0, F1, F2 and F3 cases. Conversely, differences were not significant (p > 0.05) on comparing F0, F1, F2, F3, one against the other, as well as individual versus control. Although chromosomal losses in CH were detected, it was shown that liver parenchyma with fibrosis in the initial stages (F1-F3) cannot be considered cytogenetically abnormal. PMID- 21637407 TI - Analysis of paternal lineages in Brazilian and African populations. AB - The present-day Brazilian population is a consequence of the admixture of various peoples of very different origins, namely, Amerindians, Europeans and Africans. The proportion of each genetic contribution is known to be very heterogeneous throughout the country. The aim of the present study was to compare the male lineages present in two distinct Brazilian populations, as well as to evaluate the African contribution to their male genetic substrate. Thus, two Brazilian population samples from Manaus (State of Amazon) and Ribeirao Preto (State of Sao Paulo) and three African samples from Guinea Bissau, Angola and Mozambique were typed for a set of nine Y chromosome specific STRs. The data were compared with those from African, Amerindian and European populations. By using Y-STR haplotype information, low genetic distances were found between the Manaus and Ribeirao Preto populations, as well as between these and others from Iberia. Likewise, no significant distances were observed between any of the African samples from Angola, Mozambique and Guinea Bissau. Highly significant Rst values were found between both Brazilian samples and all the African and Amerindian populations. The absence of a significant Sub-Saharan African male component resulting from the slave trade, and the low frequency in Amerindian ancestry Y-lineages in the Manaus and Ribeirao Preto population samples are in accordance with the accentuated gender asymmetry in admixture processes that has been systematically reported in colonial South American populations. PMID- 21637408 TI - A lack of association between adiponectin polymorphisms and coronary artery disease in a Chinese population. AB - We investigated the association between two single nucleotide polymorphisms (SNPs) in the adiponectin gene (rs822395 and rs266729) and coronary artery disease (CAD) in a case-control study of 198 unrelated Chinese CAD patients (with >= 70% coronary stenosis or previous myocardial infarction) and 237 non-CAD controls. The ligase reaction was used to detect SNPs rs822395 and rs266729, and the allelic association of these SNPs with the occurrence and severity of CAD was assessed. There were no significant differences in the genotypic or allelic frequencies of the two SNPs between control and CAD individuals. In addition, there was no association between the two SNPs and the severity of CAD based on the number of diseased vessels. The frequencies of alleles C and G at rs266729 differed significantly between females in the CAD and control groups, but not between males. Female carriers of allele G at rs266729 had a higher risk of CAD compared with allele C carriers (OR = 1.30, 95% CI: 1.09-2.64, p = 0.02). These results indicate a gender-specific effect of the adiponectin gene rs266729 variant in modulating the risk of CAD in women. PMID- 21637409 TI - Polymorphisms of cytochrome P450 1A1, glutathione s-transferases M1 and T1 genes in Ouangolodougou (Northern Ivory Coast). AB - In this study, the frequencies of CYP1A1, GSTM1, and GSTT1 gene polymorphisms were determined in 133 healthy individuals from Ouangolodougou, a small rural town situated in the north of the Ivory Coast. As appeared in several published studies, ethnic differences in these frequencies have been found to play an important role in the metabolism of a relevant number of human carcinogens. In the studied sample, the frequencies of Ile/Ile (wild type), Ile/Val (heterozygous variant), and Val/Val (homozygous variant) CYP1A1 genotypes were 0.271, 0.692, and 0.037, respectively. Frequencies of GSTM1 and GSTT1 null genotypes were 0.361 and 0.331, respectively. No significant differences were noted between men and women. In contrast to published data for Africans, CYP1A1 *Val Allele frequency (0.383) was significantly high (p < 0.001) in this specific population. For the GSTT1 null genotype, no differences were found between the studied and other African populations, the contrary to what occurred for the GSTM1 null genotype in relation to Gambia and Egypt. PMID- 21637410 TI - Glutathione S-transferase mu 1 (GSTM1) and theta 1 (GSTT1) genetic polymorphisms and atopic asthma in children from Southeastern Brazil. AB - Xenobiotics can trigger degranulation of eosinophils and mast cells. In this process, the cells release several substances leading to bronchial hyperactivity, the main feature of atopic asthma (AA). GSTM1 and GSTT1 genes encode enzymes involved in the inactivation of these compounds. Both genes are polymorphic in humans and have a null variant genotype in which both the gene and corresponding enzyme are absent. An increased risk for disease in individuals with the null GST genotypes is therefore, but this issue is controversial. The aim of this study was to investigate the influence of the GSTM1 and GSTT1 genotypes on the occurrence of AA, as well as on its clinical manifestations. Genomic DNA from 86 patients and 258 controls was analyzed by polymerase chain reaction. The frequency of the GSTM1 null genotype in patients was higher than that found in controls (60.5% versus 40.3%, p = 0.002). In individuals with the GSTM1 null genotype the risk of manifested AA was 2.3-fold higher (95%CI: 1.4-3.7) than for others. In contrast, similar frequencies of GSTT1 null and combined GSTM1 plus GSTT1 null genotypes were seen in both groups. No differences in genotype frequencies were perceived in patients stratified by age, gender, ethnic origin, and severity of the disease. These results suggest that the inherited absence of the GSTM1 metabolic pathway may alter the risk of AA in southeastern Brazilian children, although this must be confirmed by further studies with a larger cohort of patients and age-matched controls from the distinct regions of the country. PMID- 21637411 TI - CTLA4CT60 gene polymorphism is not associated with differential susceptibility to pemphigus foliaceus. AB - Pemphigus foliaceus is an organ-specific autoimmune disease characterized by autoantibodies against the extracellular region of desmoglein 1, a protein that mediates intercellular adhesion in desmosomes. Cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) is a key negative regulator of the T cell immune response, playing an important role in T cell homeostasis and maintenance of peripheral tolerance. Polymorphisms in the CTLA4 gene have been associated with autoimmune diseases and the functional CT60 single nucleotide polymorphism (rs3087243, also named 6230G > A) has been proposed to be a casual variant in several of these diseases. The aim of this study was to ascertain whether this polymorphism is associated with inter-individual variation in susceptibility to pemphigus foliaceus. The population sample in this case-control association study comprised 248 patient and 367 controls. We did not found a significant association of pemphigus foliaceus with the CT60 variants. We conclude that the CTLA4CT60 polymorphism is not an important factor for pemphigus foliaceus pathogenesis in the population analyzed. PMID- 21637412 TI - Familial congenital cyanosis caused by Hb-M(Yantai)(alpha-76 GAC -> TAC, Asp -> Tyr). AB - Methemoglobin (Hb-M) is a rare hemoglobinopathy in China. We hereby report on a family living in Yantai, East China, with congenital cyanosis due to Hb-M mutation. The proband, a 65-year-old female, presented 63% oxygen saturation. Both Hb-M concentration and arterial oxygen saturation remained unchanged, even following intravenous treatment with methylene blue. There was also no change in blood-color (chocolate-brown) after adding 0.1% KCN. A fast-moving band (Hb-X) in hemolysates was found by cellulose acetate electrophoresis, the Hb-X/Hb-A ratio exceeding 10%. GT transition at 131nt of exon 2, although present in one of the alpha(2) -globin alleles, was not found in alpha(1) -globin alleles as a whole. This mutation leads to the aspartic acid to tyrosine substitution (Asp76Tyr). In this family, the novel mutation in the alpha(2) -globin gene resulted in a rare form of congenital cyanosis due to Hb-M. This hemoglobin was named Hb-M (Yantai) . PMID- 21637413 TI - Replication of TCF7L2 rs7903146 association with type 2 diabetes in an Iranian population. AB - The transcription factor 7-like 2 gene (TCF7L2) rs7903146 T allele is constantly associated with Type 2 diabetes in various populations and ethnic groups. Nevertheless, this has not been observed in two studies involving Arab populations. The aim of the present study was to investigate the association between TCF7L2 rs7903146 in an Iranian population. Type 2 diabetes patients (N = 258) and normal healthy control subjects (N = 168) from the same area, were examined. The ARMS- PCR (Amplification Refractory Mutation System) technique, subsequently validated by direct sequencing, was used for genotyping. Allele and genotype frequencies were significantly different between patients and controls TT vs. CT + CC [p 0.0081 OR 3.4 95%CI (1.27-11.9)] and T vs. C allele [p 0.02 OR 1.4 95%CI (1.03-1.9)]. Our data thus confirm the association between the rs7903146 T allele and T2D in an Iranian population, contrary to previous reports in Arab populations. This can possibly be attributed to differences in ethnic background or the effects of environmental factors. PMID- 21637414 TI - Molecular forms of butyrylcholinesterase and obesity. AB - This study compared obese (N = 134) and unobese (N = 92) male blood donors, regarding the relative intensity (RI) and activity of different molecular forms (G1, G2, G4 and G1-ALB) of butyrylcholinesterase (BChE, EC 3.1.1.8) found in plasma, thereby searching for an association between these variables with obesity and SNPs of exons 1 and 4 of the BCHE gene. It was shown that obese and unobese individuals do not differ in the RI of each BChE band, even when classifying the sample into three genotypes of exons 1 and 4 of the BCHE gene (-116GG/539AA, 116GG/539AT, -116GA/539AT). Although the mean BChE activity of each band was significantly higher in obese than in unobese blood donors, the proportions of BChE bands were maintained, even under the metabolic stress associated to obesity, thereby leading to infer that this proportion is somehow regulated, and may therefore be important for BChE functions. PMID- 21637415 TI - PENCALC: A program for penetrance estimation in autosomal dominant diseases. AB - We present a computer program developed for estimating penetrance rates in autosomal dominant diseases by means of family kinship and phenotype information contained within the pedigrees. The program also determines the exact 95% credibility interval for the penetrance estimate. Both executable (PenCalc for Windows) and web versions (PenCalcWeb) of the software are available. The web version enables further calculations, such as heterozygosity probabilities and assessment of offspring risks for all individuals in the pedigrees. Both programs can be accessed and down-loaded freely at the home-page address http://www.ib.usp.br/~otto/software.htm. PMID- 21637416 TI - Co-localization of GSTP1 and JNK in transitional cell carcinoma of urinary bladder. AB - Transitional cell carcinoma (TCC) of urinary bladder belongs to glutathione S transferase P1 (GSTP1) overexpressing tumors. Upregulated GSTP1 in TCC is related to apoptosis inhibition. This antiapoptotic effects of GSTP1 might be mediated through protein:protein interaction with c-Jun NH(2) -terminal kinase (JNK). Herein, we analyzed whether a direct link between GSTP1 and JNK exists in TCC. The presence of GSTP1/JNK complexes was analyzed by immunoprecipitation and Western blotting in 20 TCC specimens, obtained after surgery. Co-localization of GSTP1 and JNK was also investigated in the 5637 TCC cell line by immunofluorescence confocal microscopy. By means of immunoprecipitation we show for the first time the presence of GSTP1/JNK complexes in all TCC samples studied. A co-localization of GSTP1 and JNK was also demonstrated in the 5637 TCC cell line by means of confocal microscopy. Protein-protein interactions, together with co-localization between GSTP1 and JNK provide evidence that GSTP1 most probably inhibits apoptosis in TCC cells by non-covalent binding to JNK. PMID- 21637417 TI - Effect of polymorphisms in the Slc11a1 coding region on resistance to brucellosis by macrophages in vitro and after challenge in two Bos breeds (Blanco Orejinegro and Zebu). AB - The resistance/susceptibility of selected cattle breeds to brucellosis was evaluated in an F1 population generated by crossing animals classified as resistant (R) and susceptible (S) (R x R, R x S, S x R, S x S) based on challenges in vitro and in vivo. The association between single nucleotide polymorphisms identified in the coding region of the Slc11a1 gene and resistance/susceptibility was estimated. The trait resistance or susceptibility to brucellosis, evaluated by a challenge in vitro, showed a high heritable component in terms of additive genetic variance (h(2) = 0.54 +/- 0.11). In addition, there was a significant association (p < 0.05) between the control of bacterial survival and two polymorphisms (a 3'UTR and SNP4 located in exon 10). The antibody response of animals classified as resistant to infection by Brucella abortus differed significantly (p < 0.05) from that of susceptible animals. However, there was no significant association between single nucleotide polymorphisms located in the Slc11a1 gene and the antibody response stimulated by a challenge in vivo. PMID- 21637418 TI - Cloning, mapping and molecular characterization of porcine progesterone receptor membrane component 2 (PGRMC2) gene. AB - Progesterone plays an important role in sow reproduction by stimulating classic genomic pathways via nuclear receptors and non-genomic pathways via membrane receptors such a progesterone receptor membrane component 2 (PGRMC2). In this work, we used radiation hybrid mapping to assign PGRMC2 to pig chromosome 8 and observed that this receptor has two transcripts in pigs. The full-length cDNA of the large transcript is 1858 bp long and contains a 669-bp open reading frame (ORF) encoding a protein of 223 amino acids. The shorter transcript encodes a protein of 170 amino acids. The porcine PGRMC2 gene consists of three exons 446 bp, 156 bp and 1259 bp in length. The promoter sequence is GC-rich and lacks a typical TATA box. Several putative cis-regulatory DNA motifs were identified in the 208-bp upstream genomic region. Five single nucleotide polymorphisms (SNPs) were detected in introns* and the 3' UTR. RT-PCR indicated that the PGRMC2 gene is expressed ubiquitously in all pig tissues examined. PMID- 21637419 TI - Phenotypic variation of erythrocyte linker histone H1.c in a pheasant (Phasianus colchicus L.) population. AB - Our goal was to characterize a phenotypic variation of the pheasant erythrocyte linker histone subtype H1.c. By using two-dimensional polyacrylamide gel electrophoresis three histone H1.c phenotypes were identified. The differently migrating allelic variants H1.c1 and H1.c2 formed either two homozygous phenotypes, c1 and c2, or a single heterozygous phenotype, c1c2. In the pheasant population screened, birds with phenotype c2 were the most common (frequency 0.761) while individuals with phenotype c1 were rare (frequency 0.043). PMID- 21637420 TI - Description of the karyotype of Rhagomys rufescens Thomas, 1886 (Rodentia, Sigmodontinae) from Southern Brazil Atlantic forest. AB - Rhagomys rufescens (Rodentia: Sigmodontinae) is an endemic species of the Atlantic forest from Southern and Southeastern Brazil. Some authors consider Rhagomys as part of the tribe Thomasomyini; but its phylogenetic relationships remain unclear. Chromosomal studies on eight specimens of Rhagomys rufescens revealed a diploid number of 2n = 36 and a number of autosome arms FN = 50. GTG, CBG and Ag-NOR banding and CMA(3) /DAPI staining were performed on metaphase chromosomes. Eight biarmed and nine acrocentric pairs were found in the karyotype of this species. The X and Y chromosomes were both acrocentric. Most of the autosomes and the sex chromosomes showed positive C-bands in the pericentromeric region. The X chromosome showed an additional heterochromatic block in the proximal region of the long arm. Nucleolus organizer regions (NORs) were located in the pericentromeric region of three biarmed autosomes (pairs 4, 6 and 8) and in the telomeric region of the short arm of three acrocentrics (pairs 10, 12 and 17). CMA (3) /DAPI staining produced fluorescent signals in many autosomes, especially in pairs 4, 6, and 8. This study presents cytogenetic data of Rhagomys rufescens for the first time. PMID- 21637421 TI - Evaluation of reference genes for real-time PCR studies of Brazilian Somalis sheep infected by gastrointestinal nematodes. AB - Precise normalization with reference genes is necessary, in order to obtain reliable relative expression data in response to gastrointestinal nematode infection. By using sheep from temperate regions as models, three reference genes, viz., ribosomal protein LO (RPLO), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and succinate dehydrogenase complex subunit A (SDHA), were investigated in the abomasum, abomasal lymph nodes and small intestine of Brazilian Somalis sheep, either resistant or susceptible to gastrointestinal nematodes infections. Real time PCR was carried out by using SYBR Green I dye, and gene stability was tested by geNorm. RPLO was an ideal reference gene, since its expression was constant across treatments, presented lower variation, and was ranked as the most stable in abomasum and lymph node tissues. On the other hand, SDHA was the most stable in the small intestine followed by RPLO and GAPDH. These findings demonstrate the importance of correctly choosing reference genes prior to relative quantification. In addition, we determined that reference genes used in sheep from temperate regions, when properly tested, can be applied in animals from tropical regions such as the Brazilian Somalis sheep. PMID- 21637422 TI - Detection of diploid males in a natural colony of the cleptobiotic bee Lestrimelitta sp (Hymenoptera, Apidae). AB - When working at quantifying the genome size of stingless bees, it was observed that males of Lestrimelitta sp possessed the same amount of nuclear DNA as the females. Thus, we used flow cytometry (FCM) and cytogenetic analysis to confirm the ploidy of these individuals. The males analyzed proved to be diploid, since, through cytometric analysis, it was demonstrated that the mean genome size of both males and females was the same (C = 0.463 pg), and, furthermore, cytogenetic analysis demonstrated that both had 2n = 28 chromosomes. PMID- 21637423 TI - Karyotypic description of the stingless bee Oxytrigona cf. flaveola (Hymenoptera, Apidae, Meliponina) of a colony from Tangara da Serra, Mato Grosso State, Brazil. AB - The aim was to broaden knowledge on the cytogenetics of the subtribe Meliponina, by furnishing cytogenetic data as a contribution to the characterization of bees from the genus Oxytrigona. Individuals of the species Oxytrigona cf. flaveola, members of a colony from Tangara da Serra, Mato Grosso State, Brazil, were studied. The chromosome number was 2n = 34, distributed among four chromosomal morphologies, with the karyotype formula 8m+8sm+16st+2t. Size heteromorphism in the first metacentric pair, subsequently confirmed by sequential staining with fluorochrome (DA/DAPI/CMA(3) ), was apparent in all the examined individuals The nucleolar organizing regions (NORs) are possibly located in this metacentric chromosome pair. These data will contribute towards a better understanding of the genus Oxytrigona. Given that species in this group are threatened, the importance of their preservation and conservation can be shown in a sensible, concise fashion through studies such as this. PMID- 21637424 TI - Linkage analysis between dominant and co-dominant makers in full-sib families of out-breeding species. AB - As high-throughput genomic tools, such as the DNA microarray platform, have lead to the development of novel genotyping procedures, such as Diversity Arrays Technology (DArT) and Single Nucleotide Polymorphisms (SNPs), it is likely that, in the future, high density linkage maps will be constructed from both dominant and co-dominant markers. Recently, a strictly genetic approach was described for estimating recombination frequency (r) between co-dominant markers in full-sib families. The complete set of maximum likelihood estimators for r in full-sib families was almost obtained, but unfortunately, one particular configuration involving dominant markers, segregating in a 3:1 ratio and co-dominant markers, was not considered. Here we add nine further estimators to the previously published set, thereby making it possible to cover all combinations of molecular markers with two to four alleles (without epistasis) in a full-sib family. This includes segregation in one or both parents, dominance and all linkage phase configurations. PMID- 21637425 TI - Development of microsatellite markers for identifying Brazilian Coffea arabica varieties. AB - Microsatellite markers, also known as SSRs (Simple Sequence Repeats), have proved to be excellent tools for identifying variety and determining genetic relationships. A set of 127 SSR markers was used to analyze genetic similarity in twenty five Coffea arabica varieties. These were composed of nineteen commercially important Brazilians and six interspecific hybrids of Coffea arabica, Coffea canephora and Coffealiberica. The set used comprised 52 newly developed SSR markers derived from microsatellite enriched libraries, 56 designed on the basis of coffee SSR sequences available from public databases, 6 already published, and 13 universal chloroplast microsatellite markers. Only 22 were polymorphic, these detecting 2-7 alleles per marker, an average of 2.5. Based on the banding patterns generated by polymorphic SSR loci, the set of twenty-five coffee varieties were clustered into two main groups, one composed of only Brazilian varieties, and the other of interspecific hybrids, with a few Brazilians. Color mutants could not be separated. Clustering was in accordance with material genealogy thereby revealing high similarity. PMID- 21637426 TI - Association mapping for yield and grain quality traits in rice (Oryza sativa L.). AB - Association analysis was applied to a panel of accessions of Embrapa Rice Core Collection (ERiCC) with 86 SSR and field data from two experiments. A clear subdivision between lowland and upland accessions was apparent, thereby indicating the presence of population structure. Thirty-two accessions with admixed ancestry were identified through structure analysis, these being discarded from association analysis, thus leaving 210 accessions subdivided into two panels. The association of yield and grain-quality traits with SSR was undertaken with a mixed linear model, with markers and subpopulation as fixed factors, and kinship matrix as a random factor. Eight markers from the two appraised panels showed significant association with four different traits, although only one (RM190) maintained the marker-trait association across years and cultivation. The significant association detected between amylose content and RM190 was in agreement with previous QTL analyses in the literature. Herein, the feasibility of undertaking association analysis in conjunction with germplasm characterization was demonstrated, even when considering low marker density. The high linkage disequilibrium expected in rice lines and cultivars facilitates the detection of marker-trait associations for implementing marker assisted selection, and the mining of alleles related to important traits in germplasm. PMID- 21637427 TI - Genetic control of Eucalyptus urophylla and E. grandis resistance to canker caused by Chrysoporthe cubensis. AB - Chrysophorte cubensis induced canker occurs in nearly all tropical and subtropical regions where eucalypts are planted, causing losses in both wood quality and volume productivity, especially so in the warmer and more humid regions of Brazil. The wide inter and intra-specific genetic variability of resistance to canker among Eucalyptus species facilitates the selection of resistant plants. In this study, we evaluated resistance to this pathogen in five Eucalyptus grandis (G) and 15 E. urophylla (U) trees, as well as in 495 individuals from 27 progenies derived from crosses between the trees. In the field, six-months-old test seedlings were inoculated with C. cubensis. Lesion length in the xylem and bark was measured eight months later. The results demonstrated that xylem lesions could preferentially be used for the selection of resistant clones. Eight trees (7 U and 1 G) were susceptible, and the remainder (8 U and 4 G) resistant. Individual narrow and broad sense heritability estimates were 17 and 81%, respectively, thereby suggesting that canker resistance is quantitative and highly dependent on dominance and epistasis. PMID- 21637428 TI - Genetic diversity in natural populations of Jacaranda decurrens Cham. determined using RAPD and AFLP markers. AB - Jacaranda decurrens (Bignoniaceae) is an endemic species of the Cerrado with validated antitumoral activity. The genetic diversity of six populations of J. decurrens located in the State of Sao Paulo was determined in this study by using molecular markers for randomly amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP). Following optimization of the amplification reaction, 10 selected primers generated 78 reproducible RAPD fragments that were mostly (69.2%) polymorphic. Two hundred and five reproducible AFLP fragments were generated by using four selected primer combinations; 46.3% of these fragments were polymorphic, indicating a considerable level of genetic diversity. Analysis of molecular variance (AMOVA) using these two groups of markers indicated that variability was strongly structured amongst populations. The unweighted pair group method with arithmatic mean (UPGMA) and Pearson's correlation coefficient (RAPD -0.16, p = 0.2082; AFLP 0.37, p = 0.1006) between genetic matrices and geographic distances suggested that the population structure followed an island model in which a single population of infinite size gave rise to the current populations of J. decurrens, independently of their spatial position. The results of this study indicate that RAPD and AFLP markers were similarly efficient in measuring the genetic variability amongst natural populations of J. decurrens. These data may be useful for developing strategies for the preservation of this medicinal species in the Cerrado. PMID- 21637429 TI - Characterization of 13 microsatellite loci developed from Meconopsis horridula. AB - Meconopsis horridula is one of the eight most famous flowers in Chinese province of Yunnan. In this study, a modified biotin-streptavidin capture method was used to detect 13 microsatellite markers in the genome of M. horridula. The polymorphism of each locus was assessed in 24 samples collected from four populations. The number of alleles per locus ranged from 2 to 7 (mean: 3.2). The observed and expected heterozygosities ranged from 0.0833 to 0.9167 and 0.0816 to 0.8050, respectively. Additionally, nine of the 13 microsatellite markers were successfully amplified in three other congeneric species. These polymorphic SSR markers could be useful for studying the population genetics of M. horridula and for assessing genetic variation in this and congenerc species in conservation programs. PMID- 21637430 TI - Evaluation of extracts from Coccoloba mollis using the Salmonella/microsome system and in vivo tests. AB - The common everyday use of medicinal plants is an ancient, and still very widespread practice, whereby the need for studies on their possible toxicity and mutagenic properties. The species Coccoloba mollis has been much used in phytotherapy, mainly in cases involving loss of memory and stress. In order to investigate its genotoxic and mutagenic potential, ethanolic extracts from the leaves and roots underwent Salmonella/microsome assaying (TA98 and TA100 strains, with and without exogenous metabolism - S9), besides comet and micronucleus tests in vivo.There was no significant increase in the number of revertants/plate of Salmonella strains in any of the analyzed root-extract concentrations, although the extract itself was extremely toxic to the Salmonella TA98 strain in the tests carried out with S9 (doses varying from 0.005 to 0.5 MUg/plate). On the other hand, the leaf-extract induced mutations in the TA98 strain in the absence of S9 in the highest concentration evaluated, although at very low mutagenic potency (0.004 rev/ MUg). Furthermore, there was no statistically significant increase in the number of comets and micronuclei, in treatments involving Swiss mice. It was obvious that extracts of Coccoloba mollis, under the described experimental conditions, are not mutagenic. PMID- 21637431 TI - Genotoxicity and mutagenicity of Echinodorus macrophyllus (chapeu-de-couro) extracts. AB - Echinodorus macrophyllus, commonly known as chapeu-de-couro, is a medicinal plant used in folk medicine to treat inflammation and rheumatic diseases. In this work, we used short-term bacterial assays based on the induction of SOS functions to examine the genotoxicity and mutagenicity of an aqueous extract of E. macrophyllus leaves. Whole extract and an ethyl acetate fraction showed similar genotoxicity and caused an ~70-fold increase in lysogenic induction. The extract also gave a positive result in the SOS chromotest with an increase of 12-fold in beta-Galactosidase enzymatic units. There was a strong trend towards base substitutions and frameshifts at purine sites in the mutations induced by the extract in Escherichia coli (CC103 and CC104 strains) and Salmonella typhimurium test strains (22-fold increase in histidine revertants in TA98 strain). Since reactive oxygen species may be implicated in aging process and in degenerative diseases, we used antioxidant compounds as catalase, thiourea and dipyridyl in the lysogenic induction test. All this compounds were able to reduce the induction factor observed in the treatment with chapeu-de-couro, thus suggesting that the genotoxicity and mutagenicity were attributable to the production of reactive oxygen species that targeted DNA purines. PMID- 21637432 TI - Genotoxicity of lapachol evaluated by wing spot test of Drosophila melanogaster. AB - This study investigated the genotoxicity of Lapachol (LAP) evaluated by wing spot test of Drosophila melanogaster in the descendants from standard (ST) and high bioactivation (HB) crosses. This assay detects the loss of heterozygosity of marker genes expressed phenotypically on the fly's wings. Drosophila has extensive genetic homology to mammals, which makes it a suitable model organism for genotoxic investigations. Three-day-old larvae from ST crosses (females flr(3)/TM3, Bd(s) x males mwh/mwh), with basal levels of the cytochrome P450 and larvae of high metabolic bioactivity capacity (HB cross) (females ORR; flr(3)/TM3, Bd(s) x males mwh/mwh), were used. The results showed that LAP is a promutagen, exhibiting genotoxic activity in larvae from the HB cross. In other words, an increase in the frequency of spots is exclusive of individuals with a high level of the cytochrome P450. The results also indicate that recombinogenicity is the main genotoxic event induced by LAP. PMID- 21637433 TI - Molecular identification, phylogeny and geographic distribution of Brazilian mangrove oysters (Crassostrea). AB - Oysters (Ostreidae) manifest a high degree of phenotypic plasticity, whereby morphology is of limited value for species identification and taxonomy. By using molecular data, the aim was to genetically characterize the species of Crassostrea occurring along the Brazilian coast, and phylogenetically relate these to other Crassostrea from different parts of the world. Sequencing of the partial cytochrome oxidase c subunit I gene (COI), revealed a total of three species of Crassostrea at 16 locations along the Brazilian coast. C. gasar was found from Curuca (Para state) to Santos (Sao Paulo state), and C. rhizophorae from Fortim (Ceara state) to Florianopolis (Santa Catarina state), although small individuals of the latter species were also found at Ajuruteua beach (municipality of Braganca, Para state). An unidentified Crassostrea species was found only on Canela Island, Braganca. Crassostrea gasar and C. rhizophorae grouped with C. virginica, thereby forming a monophyletic Atlantic group, whereas Crassostrea sp. from Canela Island was shown to be more similar to Indo-Pacific oysters, and either arrived in the Atlantic Ocean before the convergence of the Isthmus of Panama or was accidentally brought to Brazil by ship. PMID- 21637434 TI - MHC class I loci of the Bar-Headed goose (Anser indicus). AB - MHC class I proteins mediate functions in anti-pathogen defense. MHC diversity has already been investigated by many studies in model avian species, but here we chose the bar-headed goose, a worldwide migrant bird, as a non-model avian species. Sequences from exons encoding the peptide-binding region (PBR) of MHC class I molecules were isolated from liver genomic DNA, to investigate variation in these genes. These are the first MHC class I partial sequences of the bar headed goose to be reported. A preliminary analysis suggests the presence of at least four MHC class I genes, which share great similarity with those of the goose and duck. A phylogenetic analysis of bar-headed goose, goose and duck MHC class I sequences using the NJ method supports the idea that they all cluster within the anseriforms clade. PMID- 21637435 TI - Analysis of the association between spawning time QTL markers and the biannual spawning behavior in rainbow trout (Oncorhynchus mykiss). AB - The rainbow trout is a salmonid fish that occasionally exhibits broodstocks with biannual spawning behavior, a phenomenon known as a double annual reproductive cycle (DARC). Spawning time quantitative trait loci (SPT-QTLs) affect the time of the year that female rainbow trout spawn and may influence expression of the DARC trait. In this study, microsatellite markers linked and unlinked to SPT-QTLs were genotyped to investigate the underlying genetics of this trait. SPT-QTLs influenced the DARC trait since in two case-control comparisons three linked markers (OmyFGT12TUF, One3ASC and One19ASC) had significant levels of allelic frequency differentiation and marker-character association. Furthermore, alleles of One3ASC and One19ASC had significantly higher frequencies in populations that carried the DARC trait. PMID- 21637436 TI - The enigmatic monotypic crab plover Dromas ardeola is closely related to pratincoles and coursers (Aves, Charadriiformes, Glareolidae). AB - The phylogenetic placement of the monotypic crab plover Dromasardeola (Aves, Charadriiformes) remains controversial. Phylogenetic analysis of anatomical and behavioral traits using phenetic and cladistic methods of tree inference have resulted in conflicting tree topologies, suggesting a close association of Dromas to members of different suborders and lineages within Charadriiformes. Here, we revisited the issue by applying Bayesian and parsimony methods of tree inference to 2,012 anatomical and 5,183 molecular characters to a set of 22 shorebird genera (including Turnix). Our results suggest that Bayesian analysis of anatomical characters does not resolve the phylogenetic relationship of shorebirds with strong statistical support. In contrast, Bayesian and parsimony tree inference from molecular data provided much stronger support for the phylogenetic relationships within shorebirds, and support a sister relationship of Dromas to Glareolidae (pratincoles and coursers), in agreement with previously published DNA-DNA hybridization studies. PMID- 21637437 TI - American marsupials chromosomes: Why study them? AB - Marsupials, one of the three main groups of mammals, are only found in Australia and in the American continent. Studies performed in Australian marsupials have demonstrated the great potential provided by the group for the understanding of basic genetic mechanisms and chromosome evolution in mammals. Genetic studies in American marsupials are relatively scarce and cytogenetic data of most species are restricted to karyotype descriptions, usually without banding patterns. Nevertheless, the first marsupial genome sequenced was that of Monodelphis domestica, a South American species. The knowledge about mammalian genome evolution and function that resulted from studies on M. domestica is in sharp contrast with the lack of genetic data on most American marsupial species. Here, we present an overview of the chromosome studies performed in marsupials with emphasis on the South American species. PMID- 21637438 TI - Effects of L1-ORF2 fragments on green fluorescent protein gene expression. AB - The retrotransposon known as long interspersed nuclear element-1 (L1) is 6 kb long, although most L1s in mammalian and other eukaryotic cells are truncated. L1 contains two open reading frames, ORF1 and ORF2, that code for an RNA-binding protein and a protein with endonuclease and reverse transcriptase activities, respectively. In this work, we examined the effects of full length L1-ORF2 and ORF2 fragments on green fluorescent protein gene (GFP) expression when inserted into the pEGFP-C1 vector downstream of GFP. All of the ORF2 fragments in sense orientation inhibited GFP expression more than when in antisense orientation, which suggests that small ORF2 fragments contribute to the distinct inhibitory effects of this ORF on gene expression. These results provide the first evidence that different 280-bp fragments have distinct effects on the termination of gene transcription, and that when inserted in the antisense direction, fragment 280-9 (the 3' end fragment of ORF2) induces premature termination of transcription that is consistent with the effect of ORF2. PMID- 21637439 TI - RNAi-mediated silencing of the Bmi-1 gene causes growth inhibition and enhances doxorubicin-induced apoptosis in MCF-7 cells. AB - The oncogene Bmi-1 is a member of the Polycomb group gene family. Its expression is found to be greatly increased in a number of malignant tumors including breast cancer. This could suggest Bmi-1 as a potent therapeutic target. In this study, RNAi was introduced to down-regulate the expression of Bmi-1 in a highly malignant breast adenocarcinoma cell line, MCF-7. A thorough study of the biological behavior and chemosensitivity changes of the MCF-7 cells was carried out in context to the therapeutic potential of Bmi-1. The results obtained indicated that siRNA targeting of Bmi-1 could lead to an efficient and specific inhibition of endogenous Bmi-1 activity. The mRNA and protein expression of Bmi-1 were determined by RT-PCR and Western blot, respectively. Furthermore, silencing of Bmi-1 resulted in a drastic inhibition of the growth of MCF-7 cells as well as G(1) /S phase transition. The number of target cells was found to increase in phase G (0) /G (1) and decrease in the S phase, but no increase in the basal level of apoptosis was noticed. On the other hand, a reduction in the expression of cyclin D1 and an increase in the expression of p21 were also noticed. Silencing of Bmi-1 made the MCF-7 cells more sensitive to the chemotherapeutic agent doxorubicin and induced a significantly higher percentage of apoptotic cells. Here, we report on a study regarding the RNAi-mediated silencing of the Bmi-1 gene in breast cancer. PMID- 21637440 TI - Identification and detection of a novel human endogenous retrovirus-related gene, and structural characterization of its related elements. AB - Up-regulation of human endogenous retroviruses (HERVs) is associated with many diseases, including cancer. In this study, an H family HERV (HERV-H)-related gene was identified and characterized. Its spliced transcript lacks protein-coding capacity and may belong to the emerging class of noncoding RNAs (ncRNAs). The 1.3 kb RNA consisting of four exons is transcribed from an Alu element upstream of a 5.0-kb structurally incomplete HERV-H element. RT-PCR and quantitative RT-PCR results indicated that expression of this HERV-related transcript was negatively associated with colon, stomach, and kidney cancers. Its expression was induced upon treatment with DNA methylation and histone deacetylation inhibitors. A BLAT search using long terminal repeats (LTRs) identified 50 other LTR homogenous HERV H elements. Further analysis of these elements revealed that all are structurally incomplete and only five exert transcriptional activity. The results presented here recommend further investigation into a potentially functional HERV-H-related ncRNA. PMID- 21637441 TI - No contribution of GSTM1 and GSTT1 null genotypes to the risk of neutropenia due to benzene exposure in Southeastern Brazil. AB - Exposure to benzene has been associated with haematological diseases such as neutropenia (NEB) and acute myeloid leukaemia (AML). We tested whether the null genotypes of the GSTM1 and GSTT1 genes, involved in benzene inactivation, altered the risk for NEB in southeastern Brazil. Genomic DNA from 55 NEB patients and 330 controls was analysed by multiplex-polymerase chain reaction. The frequency of the GSTM1, GSTT1 and combined null genotypes was similar in patients and controls (GSTM1, 27.3% vs. 38.8%, p = 0.16; GSTT1, 25.5% vs. 19.7%, p = 0.24; GSTM1/GSTT1, 12.7% vs. 6.7%, p = 0.26; respectively). The distribution of genotype classes in NEB patients was similar to normal controls, suggesting that GSTM1 and GSTT1 null genotypes make no specific contribution to the risk of NEB. As the GSTM1 and GSTT1 null genotypes were previously associated with increased risk for AML in Brazil and elsewhere, we hypothesise that different thresholds of chemical exposure relative to distinct GSTM1 and GSTT1 genotypes may determine whether AML or NEB manifests in benzene exposed individuals from southeastern Brazil. Although indicative, our results still require support by prospective and large scale epidemiological studies, with rigorous assessment of daily chemical exposures and control of the possible contribution of other polymorphic genes involved in benzene metabolism. PMID- 21637442 TI - Hb H disease resulting from the association of an alpha-thalassemia allele [ (alpha)] with an unstable alpha-globin variant [Hb Icaria]: First report on the occurrence in Brazil. AB - Hb H Disease is caused by the loss or inactivation of three of the four functional alpha-globin genes. Patients present chronic hemolytic anemia and splenomegaly. In some cases, occasional blood transfusions are required. Deletions are the main cause of this type of thalassemia ( alpha-thalassemia). We describe here an unusual case of Hb H disease caused by the combination of a common alpha(0) deletion [-( alpha) (20.5) ] with a rare point mutation (c.427T > A), thus resulting in an elongated and unstable alpha-globin variant, Hb Icaria, (X142K), with 31 additional amino-acid residues. Very high levels of Hb H and Hb Bart's were detected in the patient's red blood cells (14.7 and 19.0%, respectively). This is the first description of this infrequent association in the Brazilian population. PMID- 21637443 TI - Polymorphism of alpha-1-antitrypsin in hematological malignancies. AB - Alpha-1-antitrypsin (AAT) or serine protease inhibitor A1 (SERPINA1) is an important serine protease inhibitor in humans. The main physiological role of AAT is to inhibit neutrophil elastase (NE) released from triggered neutrophils, with an additional lesser role in the defense against damage inflicted by other serine proteases, such as cathepsin G and proteinase 3. Although there is a reported association between AAT polymorphism and different types of cancer, this association with hematological malignancies (HM) is, as yet, unknown. We identified AAT phenotypes by isoelectric focusing (in the pH 4.2-4.9 range) in 151 serum samples from patients with HM (Hodgkins lymphomas, non-Hodgkins lymphomas and malignant monoclonal gammopathies). Healthy blood-donors constituted the control group (n = 272). The evaluated population of patients as well as the control group, were at Hardy-Weinberg equilibrium for the AAT gene (chi(2) = 4.42, d.f.11, p = 0.96 and chi(2) = 4.71, d.f.11, p = 0.97, respectively). There was no difference in the frequency of deficient AAT alleles (Pi Z and Pi S) between patients and control. However, we found a significantly higher frequency of PiM1M1 homozygote and PiM1 allele in HM patients than in control (for phenotype: f = 0.5166 and 0.4118 respectively, p = 0.037; for allele: f = 0.7020 and 0.6360 respectively, p = 0.05). In addition, PiM homozygotes in HM-patients were more numerous than in controls (59% and 48%, respectively, p = 0.044). PiM1 alleles and PiM1 homozygotes are both associated with hematological malignancies, although this is considered a functionally normal AAT variant. PMID- 21637444 TI - Three novel polymorphic microsatellite markers for the glaucoma locus GLC1B by datamining tetranucleotide repeats on chromosome 2p12-q12. AB - In order to identify new markers around the glaucoma locus GLC1B as a tool to refine its critical region at 2p11.2-2q11.2, we searched the critical region sequence obtained from the UCSC database for tetranucleotide (GATA)n and (GTCT)n repeats of at least 10 units in length. Three out of four potential microsatellite loci were found to be polymorphic, heterozygosity ranging from 64.56% to 79.59%. The identified markers are useful not only for GLC1B locus but also for the study of other disease loci at 2p11.2-2q11.2, a region with scarcity of microsatellite markers. PMID- 21637445 TI - Population analysis of xenobiotic metabolizing genes in South Brazilian Euro and Afro-descendants. AB - Individual variability in xenobiotic metabolism has been associated with susceptibility to developing complex diseases. Genes involved in xenobiotic metabolism have been evaluated in association studies; the difficulty of obtaining accurate gene frequencies in mixed populations makes interpretation of the results difficult. We sought to estimate population parameters for the cytochrome P450 and glutathione S-transferase gene families, thus contributing to studies using these genes as markers. We describe the frequencies of six genes (CYP1A1, CYP2D6, CYP2E1, GSTM1, GSTT1, and GSTP1) and estimate population parameters in 115 Euro-descendants and 196 Afro-descendants from Curitiba, South of Brazil. PCR-based methods were used for genotyping, and statistical analysis were performed by AMOVA with ARLEQUIN software. The mutant allele frequencies in the Afro-descendants and Euro-descendants, respectively, were: CYP1A1*2A = 30.1% and 15.2%; CYP2D6*4 = 14.5% and 21.5%; CYP2E1*5B = 7.9% and 5%; GSTP1*B = 37.8% and 28.3%. The null genotype frequencies were: GSTM1*0 = 36.8% and 46.1%; GSTT1*0 = 24.2% and 17.4%. PMID- 21637446 TI - Schizophrenia is not associated with the ERBB3 gene in a Han Chinese population sample: Results from case-control and family-based studies. AB - ERBB3 (v-erb-b2 erythroblastic leukemia viral oncogene homolog 3), encoding a receptor of neuregulin-1 (NRG1), has been considered a functional candidate gene for schizophrenia susceptibility. In order to investigate a relationship between ERBB3 gene and schizophrenia in the Chinese population, case-control and family based studies were carried out in 470 cases matched by controls, and in 532 family trios. Our results failed to show any evidence of significant association between the ERBB3 rs2292238 polymorphism and schizophrenia. PMID- 21637447 TI - Occurrence and abundance of a mariner-like element in freshwater and terrestrial planarians (Platyhelminthes, Tricladida) from southern Brazil. AB - Transposable elements are DNA sequences present in all the large phylogenetic groups, both capable of changing position within the genome and constituting a significant part of eukaryotic genomes. The mariner family of transposons is one of the few which occurs in a wide variety of taxonomic groups, including freshwater planarians. Nevertheless, so far only five planarian species have been reported to carry mariner-like elements (MLEs), although several different species have been investigated. Regarding the number of copies of MLEs, Girardia tigrina is the only planarian species in which this has been evaluated, with an estimation of 8,000 copies of the element per haploid genome. Preliminary results obtained in our laboratory demonstrated that MLE is found in a large number of different species of planarians, including terrestrial. With this in mind, the aim was to evaluate the occurrence and estimate the number of MLE copies in different planarian species collected in south Brazil. Twenty-eight individuals from 15 planarian species were analyzed. By using PCR and the hybridization of nucleic acids, it was found that MLE was present in all the analyzed species, the number of copies being high, probably over 10(3) per haploid genome. PMID- 21637448 TI - Cytogenetic analysis of three species of Pseudacteon (Diptera, Phoridae) parasitoids of the fire ants using standard and molecular techniques. AB - Pseudacteon flies, parasitoids of worker ants, are being intensively studied as potentially effective agents in the biological control of the invasive pest fire ant genus Solenopsis (Hymenoptera: Formicidae). This is the first attempt to describe the karyotype of P. curvatus Borgmeier, P. nocens Borgmeier and P. tricuspis Borgmeier. The three species possess 2n = 6; chromosomes I and II were metacentric in the three species, but chromosome pair III was subtelocentric in P. curvatus and P. tricuspis, and telocentric in P. nocens. All three species possess a C positive band in chromosome II, lack C positive heterochromatin on chromosome I, and are mostly differentiated with respect to chromosome III. P. curvatus and P. tricuspis possess a C positive band, but at different locations, whereas this band is absent in P. nocens. Heterochromatic bands are neither AT nor GC rich as revealed by fluorescent banding. In situ hybridization with an 18S rDNA probe revealed a signal on chromosome II in a similar location to the C positive band in the three species. The apparent lack of morphologically distinct sex chromosomes is consistent with proposals of environmental sex determination in the genus. Small differences detected in chromosome length and morphology suggests that chromosomes have been highly conserved during the evolutionary radiation of Pseudacteon. Possible mechanisms of karyotype evolution in the three species are suggested. PMID- 21637449 TI - Comparative cytogenetic analysis between Lonchorhina aurita and Trachops cirrhosus (Chiroptera, Phyllostomidae). AB - Phyllostomidae comprises the most diverse family of neotropical bats, its wide range of morphological features leading to uncertainty regarding phylogenetic relationships. Seeing that cytogenetics is one of the fields capable of providing support for currently adopted classifications through the use of several markers, a comparative analysis between two Phyllostomidae species was undertaken in the present study, with a view to supplying datasets for the further establishment of Phyllostomidae evolutionary relationships. Karyotypes of Lonchorhina aurita (2n = 32; FN = 60) and Trachops cirrhosus (2n = 30; FN = 56) were analyzed by G- and C banding, silver nitrate staining (Ag-NOR) and base-specific fluorochromes. Chromosomal data obtained for both species are in agreement with those previously described, except for X chromosome morphology in T. cirrhosus, hence indicating chromosomal geographical variation in this species. A comparison of G-banding permitted the identification of homeologies in nearly all the chromosomes. Furthermore, C-banding and Ag-NOR patterns were comparable to what has already been observed in the family. In both species CMA(3) /DA/DAPI staining revealed an R-banding-like pattern with CMA (3) , whereas DAPI showed uniform staining in all the chromosomes. Fluorochrome staining patterns for pericentromeric constitutive heterochromatin (CH) regions, as well as for nucleolar organizing regions (NORs), indicated heterogeneity regarding these sequences among Phyllostomidae species. PMID- 21637450 TI - Bayesian inference in genetic parameter estimation of visual scores in Nellore beef-cattle. AB - The aim of this study was to estimate the components of variance and genetic parameters for the visual scores which constitute the Morphological Evaluation System (MES), such as body structure (S), precocity (P) and musculature (M) in Nellore beef-cattle at the weaning and yearling stages, by using threshold Bayesian models. The information used for this was gleaned from visual scores of 5,407 animals evaluated at the weaning and 2,649 at the yearling stages. The genetic parameters for visual score traits were estimated through two-trait analysis, using the threshold animal model, with Bayesian statistics methodology and MTGSAM (Multiple Trait Gibbs Sampler for Animal Models) threshold software. Heritability estimates for S, P and M were 0.68, 0.65 and 0.62 (at weaning) and 0.44, 0.38 and 0.32 (at the yearling stage), respectively. Heritability estimates for S, P and M were found to be high, and so it is expected that these traits should respond favorably to direct selection. The visual scores evaluated at the weaning and yearling stages might be used in the composition of new selection indexes, as they presented sufficient genetic variability to promote genetic progress in such morphological traits. PMID- 21637451 TI - Genetic parameters for growth, reproductive and maternal traits in a multibreed meat sheep population. AB - The genetic parameters for growth, reproductive and maternal traits in a multibreed meat sheep population were estimated by applying the Average Information Restricted Maximum Likelihood method to an animal model. Data from a flock supported by the Programa de Melhoramento Genetico de Caprinos e Ovinos de Corte (GENECOC) were used. The traits studied included birth weight (BW), weaning weight (WW), slaughter weight (SW), yearling weight (YW), weight gain from birth to weaning (GBW), weight gain from weaning to slaughter (GWS), weight gain from weaning to yearling (GWY), age at first lambing (AFL), lambing interval (LI), gestation length (GL), lambing date (LD - number of days between the start of breeding season and lambing), litter weight at birth (LWB) and litter weight at weaning (LWW). The direct heritabilities were 0.35, 0.81, 0.65, 0.49, 0.20, 0.15 and 0.39 for BW, WW, SW, YW, GBW, GWS and GWY, respectively, and 0.04, 0.06, 0.10, 0.05, 0.15 and 0.11 for AFL, LI, GL, LD, LWB and LWW, respectively. Positive genetic correlations were observed among body weights. In contrast, there was a negative genetic correlation between GBW and GWS (-0.49) and GBW and GWY (-0.56). Positive genetic correlations were observed between AFL and LI, LI and GL, and LWB and LWW. These results indicate a strong maternal influence in this herd and the presence of sufficient genetic variation to allow mass selection for growth traits. Additive effects were of little importance for reproductive traits, and other strategies are necessary to improve the performance of these animals. PMID- 21637452 TI - A porcine gene, PBK, differentially expressed in the longissimus muscle from Meishan and Large White pig. AB - An investigation of differences in gene expression in the longissimus muscle of Meishan and Large White pigs was undertaken, using the mRNA display technique. A fragment of one differentially expressed gene was isolated and sequenced, whereupon the complete cDNA sequence was then obtained by using the rapid amplification of cDNA ends (RACE). The nucleotide sequence of the gene is not related to any known porcine gene. Sequence analysis revealed that the open reading frame of this gene encodes a protein with 322 amino acids, thus displaying high sequence identity with the PDZ binding kinase (PBK) of eleven other animal species - dog, horse, cattle, human, chimpanzee, crab-eating macaque, rhesus monkey, rat, mouse, gray short-tailed opossum and platypus, so it can be defined as the porcine PBK gene. This gene was finally assigned GeneID:100141310. Phylogenetic tree analysis revealed that the swine PBK gene has a closer genetic relationship with the PBK gene of platypus. Gene expression analysis of eight tissues of a Meishan x Large White cross showed that the porcine PBK gene is differentially expressed in various tissues. Our experiment established the primary foundation for further research on this gene. PMID- 21637453 TI - Expression profile of genes associated with mastitis in dairy cattle. AB - In order to characterize the expression of genes associated with immune response mechanisms to mastitis, we quantified the relative expression of the IL-2, IL-4, IL-6, IL-8, IL-10, IFN-gamma and TNF- alpha genes in milk cells of healthy cows and cows with clinical mastitis. Total RNA was extracted from milk cells of six Black and White Holstein (BW) cows and six Gyr cows, including three animals with and three without mastitis per breed. Gene expression was analyzed by real-time PCR. IL-10 gene expression was higher in the group of BW and Gyr cows with mastitis compared to animals free of infection from both breeds (p < 0.05). It was also higher in BW Holstein animals with clinical mastitis (p < 0.001), but it was not significant when Gyr cows with and without mastitis were compared (0.05 < p < 0.10). Among healthy cows, BW Holstein animals tended to present a higher expression of all genes studied, with a significant difference for the IL-2 and IFN- gamma genes (p < 0.001). For animals with mastitis no significant difference in gene expression was observed between the two breeds. These findings suggest that animals with mastitis develop a preferentially cell-mediated immune response. Further studies including larger samples are necessary to better characterize the gene expression profile in cows with mastitis. PMID- 21637454 TI - New occurrence of B chromosomes in Partamonahelleri (Friese, 1900) (Hymenoptera, Meliponini). AB - Cytogenetic analyses of the stingless bee Partamona helleri collected in the state of Bahia, Northeast Brazil revealed the chromosome numbers n = 18 in the haploid males and 2n = 35 in the diploid females. All karyotypes displayed one large acrocentric B chromosome, which differs from the minute B chromosomes previously described in the populations from southeastern Brazil. Giemsa staining, C-banding and DAPI/CMA(3) fluorochrome staining also revealed a remarkable interpopulational divergence regarding both the regular karyotype and the B chromosomes. The B chromosomes found in the samples from Jequie, Bahia, were entirely heterochromatic, while those found in Cravolandia, Bahia, displayed a euchromatic portion at the telomeric end of the long arm. CMA (3) labeling sites varied from seven to eight between the two localities in Bahia, due to the presence of an extra GC-rich block in the karyotype of the samples from Jequie. This is the first report of a large B chromosome in P. helleri and reveals the occurrence of a geographic differentiation within this species. PMID- 21637455 TI - Synteny of human chromosomes 14 and 15 in the platyrrhines (Primates, Platyrrhini). AB - In order to study the intra- and interspecific variability of the 14/15 association in Platyrrhini, we analyzed 15 species from 13 genera, including species that had not been described yet. The DNA libraries of human chromosomes 14 and 15 were hybridized to metaphases of Alouatta guariba clamitans, A. caraya, A. sara, Ateles paniscus chamek, Lagothrix lagothricha, Brachyteles arachnoides, Saguinus midas midas, Leontopithecus chrysomelas, Callimico goeldii, Callithrix sp., Cebus apella, Aotus nigriceps, Cacajao melanocephalus,Chiropotes satanas and Callicebus caligatus. The 14/15 hybridization pattern was present in 13 species, but not in Alouatta sara that showed a 14/15/14 pattern and Aotus nigriceps that showed a 15/14/15/14 pattern. In the majority of the species, the HSA 14 homologue retained synteny for the entire chromosome, whereas the HSA 15 homologue displayed fragmented segments. Within primates, the New World monkeys represent the taxon with the highest variability in chromosome number (2n = 16 to 62). The presence of the HSA 14/15 association in all species and subspecies studied herein confirms that this association is the ancestral condition for platyrrhines and that this association has been retained in most platyrrhines, despite the occurrence of extensive inter- and intrachromosomal rearrangements in this infraorder of Primates. PMID- 21637456 TI - Comparative cytogenetics among populations of Astyanax altiparanae (Characiformes, Characidae, Incertae sedis). AB - Cytogenetic data are presented for Astyanax altiparanae populations from three Brazilian hydrographic systems. The chromosomal data obtained in A. altiparanae support the hypothesis of diploid number conservation. However, small differences in the karyotype formula and number of nucleolar organizer regions were observed in these populations. The apparent karyotypical similarity among the studied populations strongly suggests a close relationship among them with some chromosomal divergences due to gene flow restriction. PMID- 21637457 TI - Cytological analysis of hybrids among triticales and trigopiros. AB - WE STUDIED THREE DIFFERENT TRICEPIROS: (Don Santiago x Don Noe), (Cume x Horovitz) and (Cume x Don Noe). The tricepiro (Don Santiago x Don Noe) was obtained by crossing the triticale Don Santiago INTA (AABBRR, 2n = 6x = 42) with the trigopiro Don Noe INTA (AABBDDJJ, 2n = 8x = 56). The number of chromosomes for the F(1) was 2n = 49, the most frequent meiotic configuration being 14 bivalents and 21 univalents. The univalents were situated in the periphery of the equatorial plane, whereas the bivalents were located in the central zone. The chromatids in some of the univalents split when bivalents underwent reductional division in anaphase I. There were few laggard chromosomes or chromatids at this phase. The number of chromosomes (2n = 48-58) was high and variable, and the number of bivalents per cell (18-23) also high in F (3) individuals. In all F (8) tricepiros (Don Santiago x Don Noe), F (12) tricepiros (Cume x Horovitz) and F (12) tricepiros (Cume x Don Noe), the number of chromosomes (2n = 42) was the same, these retaining the rye genome, as demonstrated by GISH and FISH. These new synthesized allopolyploids constitute interesting models for investigating the evolutionary changes responsible for diploidization, and the chromosomal and genomic re-ordering that cannot be revealed in natural allopolyploids. PMID- 21637458 TI - Altered expression of the caffeine synthase gene in a naturally caffeine-free mutant of Coffea arabica. AB - In this work, we studied the biosynthesis of caffeine by examining the expression of genes involved in this biosynthetic pathway in coffee fruits containing normal or low levels of this substance. The amplification of gene-specific transcripts during fruit development revealed that low-caffeine fruits had a lower expression of the theobromine synthase and caffeine synthase genes and also contained an extra transcript of the caffeine synthase gene. This extra transcript contained only part of exon 1 and all of exon 3. The sequence of the mutant caffeine synthase gene revealed the substitution of isoleucine for valine in the enzyme active site that probably interfered with enzymatic activity. These findings indicate that the absence of caffeine in these mutants probably resulted from a combination of transcriptional regulation and the presence of mutations in the caffeine synthase amino acid sequence. PMID- 21637459 TI - Chromosome studies in Orchidaceae from Argentina. AB - The center of diversity of Argentinean orchids is in the northeast region of the country. Chromosome numbers and karyotype features of 43 species belonging to 28 genera are presented here. Five chromosome records are the first ones at the genus level; these taxa are Aspidogyne kuckzinskii (2n = 42), Eurystyles actinosophila (2n = 56), Skeptrostachys paraguayensis (2n = 46), Stigmatosema polyaden (2n = 40) and Zygostates alleniana (2n = 54). In addition, a chromosome number is presented for the first time for 15 species: Corymborkis flava (2n = 56), Cyclopogon callophyllus (2n = 28), C. oliganthus (2n = 64), Cyrtopodium hatschbachii (2n = 46), C. palmifrons (2n = 46), Galeandra beyrichii (2n = 54), Habenaria bractescens (2n = 44), Oncidium edwallii (2n = 42), O. fimbriatum (2n = 56), O. pubes (2n = 84), O. riograndense (2n = 56), Pelexia ekmanii (2n = 46), P. lindmanii (2n = 46) and Warrea warreana (2n = 48). For Oncidium longicornu (2n = 42), O. divaricatum (2n = 56) and Sarcoglottis fasciculata (2n = 46+1B?, 46+3B?), a new cytotype was found. Chromosome data support phylogenetic relationships proposed by previous cytological, morphologic and molecular analyses, and in all the cases cover some gaps in the South American literature on orchid chromosomes. PMID- 21637460 TI - Tandem repeat distribution of gene transcripts in three plant families. AB - Tandem repeats (microsatellites or SSRs) are molecular markers with great potential for plant genetic studies. Modern strategies include the transfer of these markers among widely studied and orphan species. In silico analyses allow for studying distribution patterns of microsatellites and predicting which motifs would be more amenable to interspecies transfer. Transcribed sequences (Unigene) from ten species of three plant families were surveyed for the occurrence of micro and minisatellites. Transcripts from different species displayed different rates of tandem repeat occurrence, ranging from 1.47% to 11.28%. Both similar and different patterns were found within and among plant families. The results also indicate a lack of association between genome size and tandem repeat fractions in expressed regions. The conservation of motifs among species and its implication on genome evolution and dynamics are discussed. PMID- 21637461 TI - Genetic changes in grapevine genomes after stress induced by in vitro cultivation, thermotherapy and virus infection, as revealed by AFLP. AB - The Amplification Fragment Length Polymorphism (AFLP) technique was employed to study genetic variations which can be induced in vines by the stress occurring during different aspects of viticulture (in vitro cultivation, in vitro thermotherapy and virus infection). Analysis of AFLP banding patterns, generated by using 15 primer combinations, pointed to negligible genetic variation among plants exposed to individual stress. The average of similarity coefficients between differently stressed plants of the cultivars Muller Thurgau and Riesling were 0.984 and 0.991, respectively, as revealed by AFLP analysis. The low incidence of observed polymorphism demonstrates the high level of genome uniformity in plants reproduced by in vitro micropropagation via nodes, those subjected to in vitro thermotherapy and virus-infected plants. PMID- 21637462 TI - Distinctively variable sequence-based nuclear DNA markers for multilocus phylogeography of the soybean- and rice-infecting fungal pathogen Rhizoctonia solani AG-1 IA. AB - A series of multilocus sequence-based nuclear DNA markers was developed to infer the phylogeographical history of the Basidiomycetous fungal pathogen Rhizoctonia solani AG-1 IA infecting rice and soybean worldwide. The strategy was based on sequencing of cloned genomic DNA fragments (previously used as RFLP probes) and subsequent screening of fungal isolates to detect single nucleotide polymorphisms (SNPs). Ten primer pairs were designed based on these sequences, which resulted in PCR amplification of 200-320 bp size products and polymorphic sequences in all markers analyzed. By direct sequencing we identified both homokaryon and heterokaryon (i.e. dikaryon) isolates at each marker. Cloning the PCR products effectively estimated the allelic phase from heterokaryotic isolates. Information content varied among markers from 0.5 to 5.9 mutations per 100 bp. Thus, the former RFLP codominant probes were successfully converted into six distinctively variable sequence-based nuclear DNA markers. Rather than discarding low polymorphism loci, the combination of these distinctively variable anonymous nuclear markers would constitute an asset for the unbiased estimate of the phylogeographical parameters such as population sizes and divergent times, providing a more reliable species history that shaped the current population structure of R. solani AG-1 IA. PMID- 21637463 TI - Protective effects of solvent fractions of Mentha spicata (L.) leaves evaluated on 4-nitroquinoline-1-oxide induced chromosome damage and apoptosis in mouse bone marrow cells. AB - Spearmint leaves (Mentha spicata L.) contain high levels of antioxidants that are known to protect against both exogenous and endogenous DNA damage. In this study, the protective effects of the hexane fraction (HF), chloroform fraction (CF) and ethyl acetate fraction (EAF) in an ethanol extract from M. spicata were evaluated against 4-nitroquinoline-1-oxide (4-NQO) induced chromosome damage and apoptosis in bone marrow cells of Swiss albino mice. Two (EAF; 80 and 160 mg/ kg body weight - bw) or three (HF and CF; 80, 160 and 320 mg/ kg bw) doses of solvent fractions or vehicle control (25% DMSO in water) were administered orally for five consecutive days. Upon the sixth day, 4-NQO was injected intraperitoneally. The animals were killed the following day. Other control groups were comprised of animals treated with either the vehicle control or the various doses of solvent fractions, but with no 4-NQO treatment. 4-NQO induced micro-nucleated polychromatic erythrocytes (MnPCEs) in all the test groups. However, pre treatment of animals with the solvent fractions significantly reduced the 4-NQO induced MnPCEs as well as the percentage of apoptotic cells. The reduction of both MnPCE and apoptosis was more evident following the pre-treatment of animals with 160 mg/kg bw EAF. PMID- 21637464 TI - Toxicity and genotoxicity of wastewater from gasoline stations. AB - The toxicity and genotoxicity of wastewater from eight gasoline stations in Brasilia, Brazil's capital city, was studied by assessing chromosomal aberrations, chromosomal malsegregation and the mitotic index in Alliumcepa root cells, and the occurrence of micronucleus and nuclear abnormalities in peripheral erythrocytes of tilapia fish (Oreochromis niloticus). The content of gasoline station effluents was also analyzed based on several physico-chemical parameters. None of the wastewater samples was genotoxic to A. cepa root cells, although cell proliferation was significantly inhibited, especially at the highest concentrations. Likewise, no micronuclei were observed in O. niloticus peripheral erythrocytes, even after exposure to high concentrations, but there was an increase in the number of nuclear abnormalities and fish mortality. These results show that although the effluent from gasoline stations is processed by an oil/water separation system before being discharged into the main sewage system, the wastewater still contains toxic compounds. PMID- 21637465 TI - Genetic structure of drone congregation areas of Africanized honeybees in southern Brazil. AB - As yet, certain aspects of the Africanization process are not well understood, for example, the reproductive behavior of African and European honeybees and how the first Africanized swarms were formed and spread. Drone congregation areas (DCAs) are the ideal place to study honeybee reproduction under natural conditions since hundreds of drones from various colonies gather together in the same geographical area for mating. In the present study, we assessed the genetic structure of seven drone congregations and four commercial European-derived and Africanized apiaries in southern Brazil, employing seven microsatellite loci for this purpose. We also estimated the number of mother-colonies that drones of a specific DCA originated from. Pairwise comparison failed to reveal any population sub-structuring among the DCAs, thus indicating low mutual genetic differentiation. We also observed high genetic similarity between colonies of commercial apiaries and DCAs, besides a slight contribution from a European derived apiary to a DCA formed nearby. Africanized DCAs seem to have a somewhat different genetic structure when compared to the European. PMID- 21637466 TI - Population structure of the Monocelis lineata (Proseriata, Monocelididae) species complex assessed by phylogenetic analysis of the mitochondrial Cytochrome c Oxidase subunit I (COI) gene. AB - Monocelis lineata consists of a complex of sibling species, widespread in the Mediterranean and Atlantic Ocean. Previous genetic analysis placed in evidence at least four sibling species. Nevertheless, this research was not conclusive enough to fully resolve the complex or to infer the phylogeny/phylogeography of the group. We designed specific primers aiming at obtaining partial sequences of the mtDNA gene Cytochrome c Oxidase subunit I (COI) of M. lineata, and have identified 25 different haplotypes in 32 analyzed individuals. The dendrogram generated by Neighbor-Joining analysis confirmed the differentiation between Atlantic and Mediterranean siblings, as well as the occurrence of at least two Mediterranean sibling species. Thus validated, the method here presented appears as a valuable tool in population genetics and biodiversity surveys on the Monocelis lineata complex. PMID- 21637467 TI - Genetic variability of two populations of Pseudoplatystoma reticulatum from the Upper Paraguay River Basin. AB - Catfishes of the genus Pseudoplatystoma are very important species due to both their high commercial value and their ecological role as voracious predators. They undertake lengthy migratory movements during their life-cycle, this including reproductive migration which occurs from October to December in the rainy season. In the present study, seven microsatellite loci were analyzed to access genetic variability in two samples of P. reticulatum from the Upper Paraguay Basin. The loci were highly polymorphic (mean = 7.28). According to all analysis, the two samples of P. reticulatum revealed pronounced genetic differentiation. F(st) value was 0.2290, R(st) value 0.1067 and AMOVA 22.90% (F(st) ) and 10.67% (R(st) ), all being highly significant (p < 0.001). The division of the fishes into two groups was confirmed by microsatellite multi locus Bayesian assignment testing. The results obtained present evidence of genetic structuring in a P. reticulatum population. PMID- 21637468 TI - Parentage test in broad-snouted caimans (Caiman latirostris, Crocodylidae) using microsatellite DNA. AB - In this study, microsatellite markers, developed for Alligator mississipiensis and Caiman latirostris, were used to assess parentage among individuals from the captive colony of Caiman latirostris at the University of Sao Paulo, in Piracicaba, Sao Paulo, Brazil. Many of the females in the colony were full siblings, which made maternal identification difficult due to genotypic similarity. Even so, the most likely mother could be identified unambiguously among offspring in most of the clutches studied. Two non-parental females displayed maternal behavior which would have misled managers in assigning maternity based on behavior alone. This set of variable loci demonstrates the utility of parentage testing in captive propagation programs. PMID- 21637469 TI - Genetic structure of sigmodontine rodents (Cricetidae) along an altitudinal gradient of the Atlantic Rain Forest in southern Brazil. AB - The population genetic structure of two sympatric species of sigmodontine rodents (Oligoryzomys nigripes and Euryoryzomys russatus) was examined for mitochondrial DNA (mtDNA) sequence haplotypes of the control region. Samples were taken from three localities in the Atlantic Rain Forest in southern Brazil, along an altitudinal gradient with different types of habitat. In both species there was no genetic structure throughout their distribution, although levels of genetic variability and gene flow were high. PMID- 21637470 TI - Genomic imbalances in esophageal squamous cell carcinoma identified by molecular cytogenetic techniques. AB - This review summarizes the chromosomal changes detected by molecular cytogenetic approaches in esophageal squamous cell carcinoma (ESCC), the ninth most common malignancy in the world. Whole genome analyses of ESCC cell lines and tumors indicated that the most frequent genomic gains occurred at 1, 2q, 3q, 5p, 6p, 7, 8q, 9q, 11q, 12p, 14q, 15q, 16, 17, 18p, 19q, 20q, 22q and X, with focal amplifications at 1q32, 2p16-22, 3q25-28, 5p13-15.3, 7p12-22, 7q21-22, 8q23-24.2, 9q34, 10q21, 11p11.2, 11q13, 13q32, 14q13-14, 14q21, 14q31-32, 15q22-26, 17p11.2, 18p11.2-11.3 and 20p11.2. Recurrent losses involved 3p, 4, 5q, 6q, 7q, 8p, 9, 10p, 12p, 13, 14p, 15p, 18, 19p, 20, 22, Xp and Y. Gains at 5p and 7q, and deletions at 4p, 9p, and 11q were significant prognostic factors for patients with ESCC. Gains at 6p and 20p, and losses at 10p and 10q were the most significant imbalances, both in primary carcinoma and in metastases, which suggested that these regions may harbor oncogenes and tumor suppressor genes. Gains at 12p and losses at 3p may be associated with poor relapse-free survival. The clinical applicability of these changes as markers for the diagnosis and prognosis of ESCC, or as molecular targets for personalized therapy should be evaluated. PMID- 21637471 TI - NS3 protease from flavivirus as a target for designing antiviral inhibitors against dengue virus. AB - The development of novel therapeutic agents is essential for combating the increasing number of cases of dengue fever in endemic countries and among a large number of travelers from non-endemic countries. The dengue virus has three structural proteins and seven non-structural (NS) proteins. NS3 is a multifunctional protein with an N-terminal protease domain (NS3pro) that is responsible for proteolytic processing of the viral polyprotein, and a C-terminal region that contains an RNA triphosphatase, RNA helicase and RNA-stimulated NTPase domain that are essential for RNA replication. The serine protease domain of NS3 plays a central role in the replicative cycle of dengue virus. This review discusses the recent structural and biological studies on the NS2B-NS3 protease helicase and considers the prospects for the development of small molecules as antiviral drugs to target this fascinating, multifunctional protein. PMID- 21637472 TI - Inbreeding levels in Northeast Brazil: Strategies for the prospecting of new genetic disorders. AB - A new autosomal recessive genetic condition, the SPOAN syndrome (an acronym for spastic paraplegia, optic atrophy and neuropathy syndrome), was recently discovered in an isolated region of the State of Rio Grande do Norte in Northeast Brazil, in a population that was identified by the IBGE (Brazilian Institute of Geography and Statistics) as belonging to the Brazilian communities with the highest rates of "deficiencies" (Neri, 2003), a term used to describe diseases, malformations, and handicaps in general. This prompted us to conduct a study of consanguinity levels in five of its municipal districts by directly interviewing their inhabitants. Information on 7,639 couples (corresponding to about 40% of the whole population of the studied districts) was obtained. The research disclosed the existence of very high frequencies of consanguineous marriages, which varied from about 9% to 32%, suggesting the presence of a direct association between genetic diseases such as the SPOAN syndrome, genetic drift and inbreeding levels. This fact calls for the introduction of educational programs for the local populations, as well as for further studies aiming to identify and characterize other genetic conditions. Epidemiological strategies developed to collect inbreeding data, with the collaboration of health systems available in the region, might be very successful in the prospecting of genetic disorders. PMID- 21637473 TI - Association of homocysteine and methylene tetrahydrofolate reductase (MTHFR C677T) gene polymorphism with coronary artery disease (CAD) in the population of North India. AB - The implications of the methylene tetrahydrofolate reductase (MTHFR) gene and the level of homocysteine in the pathogenesis of coronary artery disease (CAD) have been extensively studied in various ethnic groups. Our aim was to discover the association of MTHFR (C677T) polymorphism and homocysteine level with CAD in north Indian subjects. The study group consisted of 329 angiographically proven CAD patients, and 331 age and sex matched healthy individuals as controls. MTHFR (C677T) gene polymorphism was detected based on the polymerase chain reaction and restriction digestion with HinfI. Total homocysteine plasma concentration was measured using immunoassay. T allele frequency was found to be significantly higher in patients than in the control group. We found significantly elevated levels of mean homocysteine in the patient group when compared to the control group (p = 0.00). Traditional risk factors such as diabetes, hypertension, smoking habits, a positive family history and lipid profiles (triglyceride, total cholesterol, HDL-cholesterol, LDL-cholesterol, VLDL-cholesterol), were found significantly associated through univariate analysis. Furthermore, multivariable logistics regression analysis revealed that CAD is significantly and variably associated with diabetes, hypertension, smoking, triglycerides and HDL cholesterol. Our findings showed that MTHFR C677T polymorphism and homocysteine levels were associated with coronary artery disease in the selected population. PMID- 21637474 TI - Frequencies of BCR-ABL1 fusion transcripts among Sudanese chronic myeloid leukaemia patients. AB - The incidence of one or other rearrangement in chronic myeloid leukemia (CML) patients varies in different reported series. In this study we report the frequencies of BCR-ABL1 fusion transcript variants studied in 43 CML patients from Sudan. The study includes 46 Sudanese patients, three of which negative for the BCR-ABL1 fusion transcript. More than half of 43 positive patients showed b2a2 fusion transcript (53.5%), while (41.9%) showed b3a2 transcript and the remaining (4.6%) coexpression of b3a2/ b2a2 and b3a2/b2a2/e19a2. We detected neither coexpression of p210/p190 nor e1a2 alone. Male patients showed a tendency to express b2a2, while female tende to express b3a2 (p = 0.017). Moreover, a single nucleotide polymorphism was detected in BCR exon 13 in one out of four patients and this patient showed only b2a2 expression. In conclusion, we observed a significant correlation between sex and type of BCR-ABL1 transcript, an observation that deserves further investigation. PMID- 21637475 TI - Novel mutations in the TBX5 gene in patients with Holt-Oram Syndrome. AB - The Holt-Oram syndrome (HOS) is an autosomal dominant condition characterized by upper limb and cardiac malformations. Mutations in the TBX5 gene cause HOS and have also been associated with isolated heart and arm defects. Interactions between the TBX5, GATA4 and NKX2.5 proteins have been reported in humans. We screened the TBX5, GATA4, and NKX2.5 genes for mutations, by direct sequencing, in 32 unrelated patients presenting classical (8) or atypical HOS (1), isolated congenital heart defects (16) or isolated upper-limb malformations (7). Pathogenic mutations in the TBX5 gene were found in four HOS patients, including two new mutations (c.374delG; c.678G > T) in typical patients, and the hotspot mutation c.835C > T in two patients, one of them with an atypical HOS phenotype involving lower-limb malformations. Two new mutations in the GATA4 gene were found in association with isolated upper-limb malformations, but their clinical significance remains to be established. A previously described possibly pathogenic mutation in the NKX2.5 gene (c.73C > 7) was detected in a patient with isolated heart malformations and also in his clinically normal father. PMID- 21637476 TI - Cytogenetic analysis of Astylus antis (Perty, 1830) (Coleoptera, Melyridae): Karyotype, heterochromatin and location of ribosomal genes. AB - Cytogenetic analysis of Astylus antis using mitotic and meiotic cells was performed to characterize the haploid and diploid numbers, sex determination system, chromosome morphology, constitutive heterochromatin distribution pattern and chromosomes carrying nucleolus organizer regions (NORs). Analysis of spermatogonial metaphase cells revealed the diploid number 2n = 18, with mostly metacentric chromosomes. Metaphase I cells exhibited 2n = 8II+Xyp and a parachute configuration of the sex chromosomes. Spermatogonial metaphase cells submitted to C-banding showed the presence of small dots of constitutive heterochromatin in the centromeric regions of nearly all the autosomes and on the short arm of the X chromosome (Xp), as well as an additional band on one of the arms of pair 1. Mitotic cells submitted to double staining with base-specific fluorochromes (DAPI CMA(3) ) revealed no regions rich in A+T or G+C sequences. Analysis of spermatogonial mitotic cells after sequential Giemsa/AgNO (3) staining did not reveal any specific mark on the chromosomes. Meiotic metaphase I cells stained with silver nitrate revealed a strong impregnation associated to the sex chromosomes, and in situ hybridization with an 18S rDNA probe showed ribosomal cistrons in an autosomal bivalent. PMID- 21637477 TI - Meiotic chromosomes and nucleolar behavior in testicular cells of the grassland spittlebugs Deois flavopicta, Mahanarva fimbriolata and Notozulia entreriana (Hemiptera, Auchenorrhyncha). AB - Spittlebugs annually infest pastures and cause severe damage, representing a serious problem for the tropical American beef cattle industry. Spittlebugs are an important biotic constraint to forage production and there is a lack of cytogenetic data for this group of insects. For these reasons, we conducted this work, in which the spermatogenesis and nucleolar behavior of Deois flavopicta, Mahanarva fimbriolata and Notozulia entreriana were studied. The males possessed testes in the shape of a "bunch of grapes"; a variable number of testicular lobes per individual and polyploid nuclei composed of several heteropycnotic bodies. A heteropycnotic area was located in the periphery of the nucleus (prophase I); the chiasmata were terminal or interstitial; metaphases I were circular or linear and anaphase showed late migration of the sex chromosome. The chromosome complement had 2n = 19, except for N. entreriana (2n = 15); the spermatids were round with heteropycnotic material in the center and elongated with conspicuos chromatin. The analysis of testes after silver nitrate staining showed polyploid nuclei with three large and three smaller nucleolar bodies. Early prophase cells had an intensely stained nucleolar body located close to the chromatin and another less evident body located away from the chromatin. The nucleolar bodies disintegrated during diplotene. Silver staining occurred in two autosomes, in terminal and subterminal locations, the latter probably corresponding to the nucleolus organizer regions (NORs). The spermatids were round with a round nucleolar body and silver staining was observed in the medial and posterior region of the elongated part of the spermatid head. PMID- 21637478 TI - Cytogenetic characterization of Partamona cupira (Hymenoptera, Apidae) by fluorochromes. AB - Four colonies of the stingless bee Partamona cupira (Hymenoptera: Apidae) were cytogenetically analyzed using conventional staining and the fluorochromes CMA(3) e DAPI. The females have 2n = 34 chromosomes (2K = 32 M-+2 A-). Some females, however, presented an additional large B acrocentric chromosome, to a total of 2n = 35. Chromosome B and the chromosomal pairs 2, 9 and 10 showed CMA (3) (+) bands, indicating an excess of CG base-pairs. A clear association was verified between the P. helleri B chromosome SCAR marker and the presence of a B chromosome in P. cupira. The data obtained suggests that B chromosomes in P. helleri and P. cupira share a common origin. PMID- 21637479 TI - Thaptomys Thomas 1915 (Rodentia, Sigmodontinae, Akodontini) with karyotypes 2n = 50, FN = 48, and 2n = 52, FN = 52: Two monophyletic lineages recovered by molecular phylogeny. AB - A novel karyotype with 2n = 50, FN = 48, was described for specimens of Thaptomys collected at Una, State of Bahia, Brazil, which are morphologically indistinguishable from Thaptomys nigrita, 2n = 52, FN = 52, found in other localities. It was hence proposed that the 2n = 50 karyotype could belong to a distinct species, cryptic of Thaptomys nigrita, once chromosomal rearrangements observed, along with the geographic distance, might represent a reproductive barrier between both forms. Phylogenetic analyses using maximum parsimony and maximum likelihood based on partial cytochrome b sequences with 1077 bp were performed, attempting to establish the relationships among the individuals with distinct karyotypes along the geographic distribution of the genus; the sample comprised 18 karyotyped specimens of Thaptomys, encompassing 15 haplotypes, from eight different localities of the Atlantic Rainforest. The intra-generic relationships corroborated the distinct diploid numbers, once both phylogenetic reconstructions recovered two monophyletic lineages, a northeastern clade grouping the 2n = 50 and a southeastern clade with three subclades, grouping the 2n = 52 karyotype. The sequence divergence observed between their individuals ranged from 1.9% to 3.5%. PMID- 21637480 TI - Cytogenetic analysis of three sea catfish species (Teleostei, Siluriformes, Ariidae) with the first report of Ag-NOR in this fish family. AB - Despite their ecological and economical importance, fishes of the family Ariidae are still genetically and cytogenetically poorly studied. Among the 133 known species of ariids, only eight have been karyotyped. Cytogenetic analyses performed on Genidens barbus and Sciades herzbergii revealed that both species have 2n = 56 chromosomes and Cathorops aff. mapale has 2n = 52 chromosomes: Genidens barbus has 10 Metacentrics (M), 14 Submetacentrics (SM), 26 Subtelocentrics (ST), and 6 Acrocentrics (A), Sciades herzbergii has 14M, 20SM, 18ST and 4A, whereas Cathorops aff. mapale has 14M, 20SM, and 18ST. The nucleolus organizer regions (NORs) were found in a single chromosome pair on the short arm of a large-sized ST pair in Genidens barbus and on the short arm of a middle-size SM pair in Cathorops aff. mapale. Multiple NORs on the short arms of two large sized ST pairs were found in Sciades herzbergii. The occurrence of diploid numbers ranging from 2n = 52 through 56 chromosomes and the presence of different karyotypic compositions, besides the number and position of NORs suggest that several numeric and structural chromosome rearrangements were fixed during the evolutionary history of this fish family. PMID- 21637481 TI - Association of IGF-I gene polymorphisms with milk yield and body size in Chinese dairy goats. AB - The association of IGF-I gene polymorphisms with certain traits in 708 individuals of two Chinese dairy-goat breeds (Guanzhong and Xinong Saanen) was investigated. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and DNA sequencing methods were employed in screening for genetic variation. Two novel mutations were detected in the 5'-flanking region and in intron 4 of IGF-I gene, viz., g.1617 G > A and g.5752 G > C (accession D26119.2), respectively. The associations of the g.1617 G > A mutation with milk yield and the body size were not significant (p > 0.05). However, in the case of g.5752 G > C, Xinong Saanen dairy goats with the CG genotype presented longer bodies (p < 0.05). Chest circumference (p < 0.05) was larger in Guanzhong goats with the GG genotype. In Xinong Saanen dairy goats with the CC genotype, milk yields were significantly higher during the first and second lactations (p < 0.05). Hence, the g.5752 G > C mutation could facilitate association analysis and serve as a genetic marker for Chinese dairy-goat breeding and genetics. PMID- 21637482 TI - Genetic variation in the Solanaceae fruit bearing species lulo and tree tomato revealed by Conserved Ortholog (COSII) markers. AB - The Lulo or naranjilla (Solanum quitoense Lam.) and the tree tomato or tamarillo (Solanum betaceum Cav. Sendt.) are both Andean tropical fruit species with high nutritional value and the potential for becoming premium products in local and export markets. Herein, we present a report on the genetic characterization of 62 accessions of lulos (n = 32) and tree tomatoes (n = 30) through the use of PCR based markers developed from single-copy conserved orthologous genes (COSII) in other Solanaceae (Asterid) species. We successfully PCR amplified a set of these markers for lulos (34 out of 46 initially tested) and tree tomatoes (26 out of 41) for molecular studies. Six polymorphic COSII markers were found in lulo with a total of 47 alleles and five polymorphic markers in tree tomato with a total of 39 alleles in the two populations. Further genetic analyses indicated a high population structure (with F(ST) > 0.90), which may be a result of low migration between populations, adaptation to various niches and the number of markers evaluated. We propose COSII markers as sound tools for molecular studies, conservation and the breeding of these two fruit species. PMID- 21637483 TI - Relationship between epistasis and aggressiveness in resistance of pepper (Capsicum annuum L.) to Phytophthora nicotianae. AB - This study evaluated the types of gene action governing the inheritance of resistance to Phytophthora nicotianae necrosis in populations derived from two crosses involving two susceptible (Beldi and Nabeul II) and one resistant (CM334) cultivars of pepper (Capsicum annuum L.). Populations, composed of Pr, Ps, F(1) , F (2) , BC (1) Pr, and BC (1) Ps generations, were inoculated with six P. nicotianae isolates. Generation means analysis indicated that an additive dominance model was appropriate for P. nicotianae isolates Pn (Ko1) , Pn (Ko2) and Pn (Kr1) , which showed low aggressiveness in the two crosses. For the more aggressive isolates Pn (Bz1) , Pn (Bz2) and Pn (Kr2) , epistasis was an integral component of resistance in the two crosses. The presence of epistasis in the resistance of pepper to P. nicotianae was dependent on the level of aggressiveness of the isolates. Selection in pepper with less aggressive isolates was efficient, but not with more aggressive isolates; on the other hand, selection with more aggressive isolates was more stable. The minimum number of genes controlling resistance was estimated at up to 2.71. In the majority of cases, the additive variance was significant and greater than the environmental and dominance variance. PMID- 21637484 TI - Molecular cloning and expression profiling of a chalcone synthase gene from hairy root cultures of Scutellaria viscidula Bunge. AB - A cDNA encoding chalcone synthase (CHS), the key enzyme in flavonoid biosynthesis, was isolated from hairy root cultures of Scutellaria viscidula Bunge by rapid amplification of cDNA ends (RACE). The full-length cDNA of S. viscidula CHS, designated as Svchs (GenBank accession no. EU386767), was 1649 bp with a 1170 bp open reading frame (ORF) that corresponded to a deduced protein of 390 amino acid residues, a calculated molecular mass of 42.56 kDa and a theoretical isoelectric point (pI) of 5.79. Multiple sequence alignments showed that SvCHS shared high homology with CHS from other plants. Functional analysis in silico indicated that SvCHS was a hydrophilic protein most likely associated with intermediate metabolism. The active sites of the malonyl-CoA binding motif, coumaroyl pocket and cyclization pocket in CHS of Medicago sativa were also found in SvCHS. Molecular modeling indicated that the secondary structure of SvCHS contained mainly alpha-helixes and random coils. Phylogenetic analysis showed that SvCHS was most closely related to CHS from Scutellaria baicalensis. In agreement with its function as an elicitor-responsive gene, the expression of Svchs was induced and coordinated by methyl jasmonate. To our knowledge, this is the first report to describe the isolation and expression of a gene from S. viscidula. PMID- 21637485 TI - A genomic analysis of disease-resistance genes encoding nucleotide binding sites in Sorghum bicolor. AB - A large set of candidate nucleotide-binding site (NBS)-encoding genes related to disease resistance was identified in the sorghum (Sorghum bicolor) genome. These resistance (R) genes were characterized based on their structural diversity, physical chromosomal location and phylogenetic relationships. Based on their N terminal motifs and leucine-rich repeats (LRR), 50 non-regular NBS genes and 224 regular NBS genes were identified in 274 candidate NBS genes. The regular NBS genes were classified into ten types: CNL, CN, CNLX, CNX, CNXL, CXN, NX, N, NL and NLX. The vast majority (97%) of NBS genes occurred in gene clusters, indicating extensive gene duplication in the evolution of S. bicolor NBS genes. Analysis of the S. bicolor NBS phylogenetic tree revealed two major clades. Most NBS genes were located at the distal tip of the long arms of the ten sorghum chromosomes, a pattern significantly different from rice and Arabidopsis, the NBS genes of which have a random chromosomal distribution. PMID- 21637486 TI - Restoring pollen fertility in transgenic male-sterile eggplant by Cre/loxp mediated site-specific recombination system. AB - This study was designed to control plant fertility by cell lethal gene Barnase expressing at specific developmental stage and in specific tissue of male organ under the control of Cre/loxP system, for heterosis breeding, producing hybrid seed of eggplant. The Barnase-coding region was flanked by loxP recognition sites for Cre-recombinase. The eggplant inbred/pure line ('E-38') was transformed with Cre gene and the inbred/pure line ('E-8') was transformed with the Barnase gene situated between loxp. The experiments were done separately, by means of Agrobacterium co-culture. Four T(0) -plants with the Barnase gene were obtained, all proved to be male-sterile and incapable of producing viable pollen. Flowers stamens were shorter, but the vegetative phenotype was similar to wild-type. Five T (0) -plants with the Cre gene developed well, blossomed out and set fruit normally. The crossing of male-sterile Barnase-plants with Cre expression transgenic eggplants resulted in site-specific excision with the male-sterile plants producing normal fruits. With the Barnase was excised, pollen fertility was fully restored in the hybrids. The phenotype of these restored plants was the same as that of the wild-type. Thus, the Barnase and Cre genes were capable of stable inheritance and expression in progenies of transgenic plants. PMID- 21637487 TI - Genetic divergence of rubber tree estimated by multivariate techniques and microsatellite markers. AB - Genetic diversity of 60 Hevea genotypes, consisting of Asiatic, Amazonian, African and IAC clones, and pertaining to the genetic breeding program of the Agronomic Institute (IAC), Brazil, was estimated. Analyses were based on phenotypic multivariate parameters and microsatellites. Five agronomic descriptors were employed in multivariate procedures, such as Standard Euclidian Distance, Tocher clustering and principal component analysis. Genetic variability among the genotypes was estimated with 68 selected polymorphic SSRs, by way of Modified Rogers Genetic Distance and UPGMA clustering. Structure software in a Bayesian approach was used in discriminating among groups. Genetic diversity was estimated through Nei's statistics. The genotypes were clustered into 12 groups according to the Tocher method, while the molecular analysis identified six groups. In the phenotypic and microsatellite analyses, the Amazonian and IAC genotypes were distributed in several groups, whereas the Asiatic were in only a few. Observed heterozygosity ranged from 0.05 to 0.96. Both high total diversity (H(T') = 0.58) and high gene differentiation (G (st') = 0.61) were observed, and indicated high genetic variation among the 60 genotypes, which may be useful for breeding programs. The analyzed agronomic parameters and SSRs markers were effective in assessing genetic diversity among Hevea genotypes, besides proving to be useful for characterizing genetic variability. PMID- 21637488 TI - SSR and AFLP based genetic diversity of soybean germplasm differing in photoperiod sensitivity. AB - Forty-four soybean genotypes with different photoperiod response were selected after screening of 1000 soybean accessions under artificial condition and were profiled using 40 SSR and 5 AFLP primer pairs. The average polymorphism information content (PIC) for SSR and AFLP marker systems was 0.507 and 0.120, respectively. Clustering of genotypes was done using UPGMA method for SSR and AFLP and correlation was 0.337 and 0.504, respectively. Mantel's correlation coefficients between Jaccard's similarity coefficient and the cophenetic values were fairly high in both the marker systems (SSR = 0.924; AFLP = 0.958) indicating very good fit for the clustering pattern. UPGMA based cluster analysis classified soybean genotypes into four major groups with fairly moderate bootstrap support. These major clusters corresponded with the photoperiod response and place of origin. The results indicate that the photoperiod insensitive genotypes, 11/2/1939 (EC 325097) and MACS 330 would be better choice for broadening the genetic base of soybean for this trait. PMID- 21637489 TI - A new approach for obtaining rapid uniformity in rice (Oryza sativa L.) via a 3x x 2x cross. AB - A triploid (2n = 3x = 36) rice plant was obtained by screening a twin seedling population in which each seed germinated to two or three sprouts that were then crossed with diploid plants. One diploid plant was chosen among the various F(1) progenies and developed into an F (2) population via self-pollination. Compared with the control variety Shanyou 63, this F (2) population had a stable agronomical performance in field trials, as confirmed by the F-test. The stability of the F (2) population was further substantiated by molecular analysis with simple sequence repeat markers. Specifically, of 160 markers assayed, 37 (covering all 12 chromosomes) were polymorphic between the parental lines. Testing the F (1) hybrid individually with these markers showed that each PCR product had only a single band instead of two bands from each parent. The bands were identical to either maternal (23 markers) or paternal (eight markers) bands or distinct from both parents (six markers). The amplified bands of all 60 randomly selected F (2) plants were uniform and identical to those of the F (1) hybrid. These results suggest that the F (1) plant is a non-segregating hybrid and that a stable F (2) population was obtained. This novel system provides an efficient means for shortening the cycle of hybrid rice seed production. PMID- 21637490 TI - Efficiency of IRAP and ITS-RFLP marker systems in accessing genetic variation of Pyrenophora graminea. AB - The usefulness of IRAP (inter-retrotransposon amplified polymorphism) and ITS RFLP (restriction of PCR-amplified internal transcribed spacers of the rDNA) markers in the analysis of 39 Pyrenophora graminea isolates was determined. Each marker system could discriminate between all of the isolates in detecting polymorphism, albeit with variable efficiency. IRAP and ITS-RFLP produced 85% and 77% polymorphic bands, respectively, with a corresponding mean polymorphic information content (PIC) of 0.38 and 0.36. The IRAP marker index ratio (2.41) was higher than ITS-RFLP (1.50). On one hand, the quality nature of data (QND) was higher for ITS-RFLP (0.169) than IRAP (0.093). However, correlation between both marker similarity matrices was significant (r = 0.34, p < 0.05). These findings suggest their combined use in phylogenetic analysis. To our knowledge, this is the first report of a comparison involving these two advanced DNA marker systems. PMID- 21637491 TI - PhoR/PhoP two component regulatory system affects biocontrol capability of Bacillus subtilis NCD-2. AB - The Bacillus subtilis strain NCD-2 is an important biocontrol agent against cotton verticillium wilt and cotton sore shin in the field, which are caused by Verticillium dahliae Kleb and Rhizoctonia solani Kuhn, respectively. A mutant of strain NCD-2, designated M216, with decreased antagonism to V. dahliae and R. solani, was selected by mini-Tn10 mutagenesis and in vitro virulence screening. The inserted gene in the mutant was cloned and identified as the phoR gene, which encodes a sensor kinase in the PhoP/PhoR two-component system. Compared to the wild-type strain, the APase activities of the mutant was decreased significantly when cultured in low phosphate medium, but no obvious difference was observed when cultured in high phosphate medium. The mutant also grew more slowly on organic phosphate agar and lost its phosphatidylcholine-solubilizing ability. The suppression of cotton seedling damping-off in vivo and colonization of the rhizosphere of cotton also decreased in the mutant strain when compared with the wild type strain. All of these characteristics could be partially restored by complementation of the phoR gene in the M216 mutant. PMID- 21637492 TI - Distribution and biological role of the oligopeptide-binding protein (OppA) in Xanthomonas species. AB - In this study we investigated the prevalence of the oppA gene, encoding the oligopeptide binding protein (OppA) of the major bacterial oligopeptide uptake system (Opp), in different species of the genus Xanthomonas. The oppA gene was detected in two Xanthomonas axonopodis strains among eight tested Xanthomonas species. The generation of an isogenic oppA-knockout derivative of the Xac 306 strain, showed that the OppA protein neither plays a relevant role in oligopeptide uptake nor contributes to the infectivity and multiplication of the bacterial strain in leaves of sweet orange (Citrus sinensis) and Rangpur lime (Citrus limonia). Taken together these results suggest that the oppA gene has a recent evolutionary history in the genus and does not contribute in the physiology or pathogenesis of X. axonopodis. PMID- 21637493 TI - Differential expression of pathogenicity- and virulence-related genes of Xanthomonas axonopodis pv. citri under copper stress. AB - In this study, we used real-time quantitative PCR (RT-qPCR) to evaluate the expression of 32 genes of Xanthomonas axonopodis pv. citri related to pathogenicity and virulence that are also involved in copper detoxification. Nearly all of the genes were up-regulated, including copA and copB. Two genes homologous to members of the type II secretion system (xcsH and xcsC) and two involved in the degradation of plant cell wall components (pglA and pel) were the most expressed in response to an elevated copper concentration. The type II secretion system (xcs operon) and a few homologues of proteins putatively secreted by this system showed enhanced expression when the bacteria were exposed to a high concentration of copper sulfate. The enhanced expression of the genes of secretion II system during copper stress suggests that this pathway may have an important role in the adaptative response of X. axonopodis pv. citri to toxic compounds. These findings highlight the potential role of these genes in attenuating the toxicity of certain metals and could represent an important means of bacterial resistance against chemicals used to control diseases. PMID- 21637494 TI - Molecular characterization of beta-tubulin from Phakopsora pachyrhizi, the causal agent of Asian soybean rust. AB - beta-tubulins are structural components of microtubules and the targets of benzimidazole fungicides used to control many diseases of agricultural importance. Intron polymorphisms in the intron-rich genes of these proteins have been used in phylogeographic investigations of phytopathogenic fungi. In this work, we sequenced 2764 nucleotides of the beta-tubulin gene (Pp tubB) in samples of Phakopsora pachyrhizi collected from seven soybean fields in Brazil. Pp tubB contained an open reading frame of 1341 nucleotides, including nine exons and eight introns. Exon length varied from 14 to 880 nucleotides, whereas intron length varied from 76 to 102 nucleotides. The presence of only four polymorphic sites limited the usefulness of Pp tubB for phylogeographic studies in P. pachyrhizi. The gene structures of Pp tubB and orthologous beta-tubulin genes of Melampsora lini and Uromyces viciae-fabae were highly conserved. The amino acid substitutions in beta-tubulin proteins associated with the onset of benzimidazole resistance in model organisms, especially at His (6) , Glu (198) and Phe (200) , were absent from the predicted sequence of the P. pachyrhizi beta-tubulin protein. PMID- 21637495 TI - Dietary carotenoid-rich oil supplementation improves exercise-induced anisocytosis in runners: influences of haptoglobin, MnSOD (Val9Ala), CAT (21A/T) and GPX1 (Pro198Leu) gene polymorphisms in dilutional pseudoanemia (sports anemia). AB - Physical training induces beneficial adaptation, whereas exhaustive exercises increase reactive oxygen-species generation, thereby causing oxidative damage in plasma and erythrocytes, fractions susceptible to lipid peroxidation. Pequi (Caryocar brasiliense Camb.) is a Brazilian Cerrado fruit containing a carotenoid rich oil. The aim was to investigate the effects of pequi-oil on exercise-induced oxidative damage in plasma and erythrocytes, after running in the same environment and undergoing weekly training under the same conditions as to type, intensity and length. Evaluations were accomplished after outdoor running on flat land before and after ingestion of 400 mg pequi-oil capsules for 14 days. Blood samples were taken after running and submitted to TBARS assay and erythrogram analysis. Haptoglobin, MnSOD (Val9Ala), CAT (21A/T) and GPX1 (Pro198Leu) gene polymorphisms were priorly investigated, so as to estimate genetic influence The reduction in erythrocytes, hemoglobin and hematocrit after pequi-oil treatment was notably associated with higher plasma expansion. Except for MCHC (mean corpuscular hemoglobin concentration) and RDW (red cell distribution width), the results were influenced by the polymorphisms studied. The best response to pequi oil was presented by MnSOD Val/Val, CAT AA or AT genotypes and the GPX1 Pro allele. The significantly lower RDW and higher MHCH values were related to pequi oil protective effects. Pequi oil, besides possessing other nutritional properties, showed protective blood effects. PMID- 21637496 TI - Non-homologous DNA end joining in normal and cancer cells and its dependence on break structures. AB - DNA double-strand breaks (DSBs) are a serious threat to the cell, for if not or miss-repaired, they can lead to chromosomal aberration, mutation and cancer. DSBs in human cells are repaired via non-homologous DNA end joining (NHEJ) and homologous recombination repair pathways. In the former process, the structure of DNA termini plays an important role, as does the genetic constitution of the cells, through being different in normal and pathological cells. In order to investigate the dependence of NHEJ on DSB structure in normal and cancer cells, we used linearized plasmids with various, complementary or non-complementary, single-stranded or blunt DNA termini, as well as whole-cell extract isolated from normal human lymphocytes, chronic myeloid leukemia K562 cells and lung cancer A549 cells. We observed a pronounced variability in the efficacy of NHEJ reaction depending on the type of ends. Plasmids with complementary and blunt termini were more efficiently repaired than the substrate with 3' protruding single-strand ends. The hierarchy of the effectiveness of NHEJ was on average, from the most effective to the least, A549/ normal lymphocytes/ K562. Our results suggest that the genetic constitution of the cells together with the substrate terminal structure may contribute to the efficacy of the NHEJ reaction. This should be taken into account on considering its applicability in cancer chemo- or radiotherapy by pharmacologically modulating NHEJ cellular responses. PMID- 21637497 TI - Nuclear anomalies in the buccal cells of calcite factory workers. AB - The micronucleus (MN) assay on exfoliated buccal cells is a useful and minimally invasive method for monitoring genetic damage in humans. To determine the genotoxic effects of calcite dust that forms during processing, MN assay was carried out in exfoliated buccal cells of 50 (25 smokers and 25 non-smokers) calcite factory workers and 50 (25 smokers and 25 non-smokers) age- and sex matched control subjects. Frequencies of nuclear abnormalities (NA) other than micronuclei, such as binucleates, karyorrhexis, karyolysis and 'broken eggs', were also evaluated. Micronuclei and the other aforementioned anomalies were analysed by two way analysis of covariance. The linear correlations between the types of micronucleus and nuclear abnormalities were determined by Spearman's Rho. There was a positive correlation between micronuclei and other types of nuclear abnormalities in accordance with the Spearman's Rho test. Results showed statistically significant difference between calcite fabric workers and control groups. MN and NA frequencies in calcite fabric workers were significantly higher than those in control groups (p < 0.05). The results of this study indicate that calcite fabric workers are under risk of significant cytogenetic damage. PMID- 21637498 TI - Genetic variation and evolutionary demography of Fenneropenaeus chinensis populations, as revealed by the analysis of mitochondrial control region sequences. AB - Genetic variation and evolutionary demography of the shrimp Fenneropenaeus chinensis were investigated using sequence data of the complete mitochondrial control region (CR). Fragments of 993 bp of the CR were sequenced for 93 individuals from five localities over most of the species' range in the Yellow Sea and the Bohai Sea. There were 84 variable sites defining 68 haplotypes. Haplotype diversity levels were very high (0.95 +/- 0.03-0.99 +/- 0.02) in F. chinensis populations, whereas those of nucleotide diversity were moderate to low (0.66 +/- 0.36%-0.84 +/- 0.46%). Analysis of molecular variance and conventional population statistics (F(ST) ) revealed no significant genetic structure throughout the range of F. chinensis. Mismatch distribution, estimates of population parameters and neutrality tests revealed that the significant fluctuations and shallow coalescence of mtDNA genealogies observed were coincident with estimated demographic parameters and neutrality tests, in implying important past-population size fluctuations or range expansion. Isolation with Migration (IM) coalescence results suggest that F. chinensis, distributed along the coasts of northern China and the Korean Peninsula (about 1000 km apart), diverged recently, the estimated time-split being 12,800 (7,400 18,600) years ago. PMID- 21637499 TI - Comparative study on the use of specific and heterologous microsatellite primers in the stingless bees Melipona rufiventris and M. mondury (Hymenoptera, Apidae). AB - Due to their high degree of polymorphism, microsatellites are considered useful tools for studying population genetics. Nevertheless, studies of genetic diversity in stingless bees by means of these primers have revealed a low level of polymorphism, possibly the consequence of the heterologous primers used, since in most cases these were not specifically designed for the species under consideration. Herein we compared the number of polymorphic loci and alleles per locus, as well as observed heterozygosity in Melipona rufiventris and M. mondury populations, using specific and heterologous primers. The use of specific primers placed in evidence the greater frequency of polymorphic loci and alleles per locus, besides an expressive increase in observed heterozygosity in M. rufiventris and M. mondury, thereby reinforcing the idea that populational studies should be undertaken by preferably using species-specific microsatellite primers. PMID- 21637500 TI - Variation and genetic structure of Melipona quadrifasciata Lepeletier (Hymenoptera, Apidae) populations based on ISSR pattern. AB - For a study of diversity and genetic structuring in Melipona quadrifasciata, 61 colonies were collected in eight locations in the state of Minas Gerais, Brazil. By means of PCR analysis, 119 ISSR bands were obtained, 80 (68%) being polymorphic. H(e) and H (B) were 0.20 and 0.16, respectively. Two large groups were obtained by the UPGMA method, one formed by individuals from Januaria, Urucuia, Rio Vermelho and Caete and the other by individuals from Sao Joao Del Rei, Barbacena, Ressaquinha and Cristiano Otoni. The Phist and theta(B) values were 0.65 and 0.58, respectively, thereby indicating high population structuring. UPGMA grouping did not reveal genetic structuring of M. quadrifasciata in function of the tergite stripe pattern. The significant correlation between dissimilarity values and geographic distances (r = 0.3998; p < 0.05) implies possible geographic isolation. The genetic differentiation in population grouping was probably the result of an interruption in gene flow, brought about by geographic barriers between mutually close geographical locations. Our results also demonstrate the potential of ISSR markers in the study of Melipona quadrifasciata population structuring, possibly applicable to the studies of other bee species. PMID- 21637501 TI - Comparison between simulated annealing algorithms and rapid chain delineation in the construction of genetic maps. AB - The efficiency of simulated annealing algorithms and rapid chain delineation in establishing the best linkage order, when constructing genetic maps, was evaluated. Linkage refers to the phenomenon by which two or more genes, or even more molecular markers, can be present in the same chromosome or linkage group. In order to evaluate the capacity of algorithms, four F(2) co-dominant populations, 50, 100, 200 and 1000 in size, were simulated. For each population, a genome with four linkage groups (100 cM) was generated. The linkage groups possessed 51, 21, 11 and 6 marks, respectively, and a corresponding distance of 2, 5, 10 and 20 cM between adjacent marks, thereby causing various degrees of saturation. For very saturated groups, with an adjacent distance between marks of 2 cM and in greater number, i.e., 51, the method based upon stochastic simulation by simulated annealing presented orders with distances equivalent to or lower than rapid chain delineation. Otherwise, the two methods were commensurate through presenting the same SARF distance. PMID- 21637502 TI - Translational selection on SHH genes. AB - Codon usage bias has been observed in various organisms. In this study, the correlation between SHH genes expression in some tissues and codon usage features was analyzed by bioinformatics. We found that translational selection may act on compositional features of this set of genes. PMID- 21637503 TI - Genomic rearrangements in BRCA1 and BRCA2: A literature review. AB - Women with mutations in the breast cancer genes BRCA1 or BRCA2 have an increased lifetime risk of developing breast, ovarian and other BRCA-associated cancers. However, the number of detected germline mutations in families with hereditary breast and ovarian cancer (HBOC) syndrome is lower than expected based upon genetic linkage data. Undetected deleterious mutations in the BRCA genes in some high-risk families are due to the presence of intragenic rearrangements such as deletions, duplications or insertions that span whole exons. This article reviews the molecular aspects of BRCA1 and BRCA2 rearrangements and their frequency among different populations. An overview of the techniques used to screen for large rearrangements in BRCA1 and BRCA2 is also presented. The detection of rearrangements in BRCA genes, especially BRCA1, offers a promising outlook for mutation screening in clinical practice, particularly in HBOC families that test negative for a germline mutation assessed by traditional methods. PMID- 21637504 TI - Population prevalence of hereditary breast cancer phenotypes and implementation of a genetic cancer risk assessment program in southern Brazil. AB - In 2004, a population-based cohort (the Nucleo Mama Porto Alegre - NMPOA Cohort) was started in Porto Alegre, southern Brazil and within that cohort, a hereditary breast cancer study was initiated, aiming to determine the prevalence of hereditary breast cancer phenotypes and evaluate acceptance of a genetic cancer risk assessment (GCRA) program. Women from that cohort who reported a positive family history of cancer were referred to GCRA. Of the 9218 women enrolled, 1286 (13.9%) reported a family history of cancer. Of the 902 women who attended GCRA, 55 (8%) had an estimated lifetime risk of breast cancer >= 20% and 214 (23.7%) had pedigrees suggestive of a breast cancer predisposition syndrome; an unexpectedly high number of these fulfilled criteria for Li-Fraumeni-like syndrome (122 families, 66.7%). The overall prevalence of a hereditary breast cancer phenotype was 6.2% (95%CI: 5.67-6.65). These findings identified a problem of significant magnitude in the region and indicate that genetic cancer risk evaluation should be undertaken in a considerable proportion of the women from this community. The large proportion of women who attended GCRA (72.3%) indicates that the program was well-accepted by the community, regardless of the potential cultural, economic and social barriers. PMID- 21637506 TI - Analysis of 31 STR loci in the genetic isolate of Carloforte (Sardinia, Italy). AB - The genotypes of 31 autosomal short tandem repeat loci in the population of Carloforte were analyzed, these representing a linguistic and genetic isolate located on the island of Sardinia (Italy). The markers span the entire length of chromosomes 19, 20, 21 and 22. Allele frequencies and statistical parameters were presented for all loci. Observed heterozygosity ranged from 0.279 to 0.884, and polymorphism information content from 0.552 to 0.886. All but two loci showed Hardy-Weinberg equilibrium after Bonferroni correction. The 31 short tandem repeat loci examined in the present work provide additional data on the genetic structure of the Carloforte population. PMID- 21637505 TI - Haptoglobin gene subtypes in three Brazilian population groups of different ethnicities. AB - Haptoglobin is a plasma hemoglobin-binding protein that limits iron loss during normal erythrocyte turnover and hemolysis, thereby preventing oxidative damage mediated by iron excess in the circulation. Haptoglobin polymorphism in humans, characterized by the Hp(*1) and Hp (*2) alleles, results in distinct phenotypes known as Hp1-1, Hp2-1 and Hp2-2, whose frequencies vary according to the ethnic origin of the population. The Hp(*1) allele has two subtypes, Hp (*1F) and Hp (*1S) , that also vary in their frequencies among populations worldwide. In this work, we examined the distribution frequencies of haptoglobin subtypes in three Brazilian population groups of different ethnicities. The haptoglobin genotypes of Kayabi Amerindians (n = 56), Kalunga Afro-descendants (n = 70) and an urban population (n = 132) were determined by allele-specific PCR. The Hp(*1F) allele frequency was highest in Kalunga (29.3%) and lowest in Kayabi (2.6%). The Hp(*1F)/Hp(*1S) allele frequency ratios were 0.6, 1.0 and 0.26 for the Kayabi, Kalunga and urban populations, respectively. This variation was attributable largely to the Hp(*1F) allele. However, despite the large variation in Hp(*1F) frequencies, results of F (ST) (0.0291) indicated slight genetic differentiation among subpopulations of the general Brazilian population studied here. This is the first Brazilian report of variations in the Hp(*1F) and Hp(*1S) frequencies among non-Amerindian Brazilians. PMID- 21637507 TI - Mutation and association analysis of the PVR and PVRL2 genes in patients with non syndromic cleft lip and palate. AB - Orofacial clefts (OFC; MIM 119530) are among the most common major birth defects. Here, we carried out mutation screening of the PVR and PVRL2 genes, which are both located at an OFC linkage region at 19q13 (OFC3) and are closely related to PVRL1, which has been associated with both syndromic and non-syndromic cleft lip and palate (nsCLP). We screened a total of 73 nsCLP patients and 105 non-cleft controls from the USA for variants in PVR and PVRL2, including all exons and encompassing all isoforms. We identified four variants in PVR and five in PVRL2. One non-synonymous PVR variant, A67T, was more frequent among nsCLP patients than among normal controls, but this difference did not achieve statistical significance. PMID- 21637508 TI - The karyotype of three Brazilian Terrarana frogs (Amphibia, Anura) with evidence of a new Barycholos species. AB - A recent substantial rearrangement of the 882 described eleutherodactyline frog species has considerably improved the understanding of their systematics. Nevertheless, many taxonomic aspects of the South American eleutherodactyline species remain unknown and require further investigation using morphological, cytogenetic and molecular approaches. In this work, the karyotypes of the Brazilian species Ischnocnema juipoca (Atibaia and Campos do Jordao, SP), Barycholos cf. ternetzi (Uberlandia, MG, and Porto Nacional, TO), and Pristimantis crepitans (Chapada dos Guimaraes and Sao Vicente, MT) were analyzed using Giemsa staining, Ag-NOR labeling, and C-banding techniques. All individuals had a diploid number of 22 chromosomes, but the Fundamental Numbers were different among species. The herein described low chromosome number of Pristimantis crepitans is unique within this genus, suggesting that cytogenetically this species is not closely related either to its congeneric species or to Ischnocnema. In addition, karyotype differences, mainly in the NOR position, clearly distinguished the two Barycholos populations, besides indicating the existence of a so far undescribed species in this genus. A taxonomic review could clarify the systematic position of P. crepitans and verify the hypothetic new Barycholos species. PMID- 21637509 TI - Astyanax hastatus Myers, 1928 (Teleostei, Characidae): A new species complex within the genus Astyanax? AB - Four populations of Astyanax hastatus Myers 1928 from the Guapimirim River basin (Rio de Janeiro State) were analyzed and three distinct cytotypes identified. These cytotypes presented 2n = 50 chromosomes, with 4M+8SM+10ST+28A (Cytotype A), 8M+10SM+14ST+18A (Cytotype B), 6M+8SM+4ST+32A (Cytotype C) and scanty heterochromatin, mainly located throughout pericentromeric regions of several chromosomal pairs. No homologies with the As-51 satellite DNA were observed in the three cytotypes, although all of them presented multiple 18S rDNA sites, as detected by both silver nitrate staining and FISH (fluorescent in situ hybridization). The application of the term "species complex" in Astyanax is discussed from a cytotaxonomic viewpoint. PMID- 21637510 TI - Aromatase gene and its effects on growth, reproductive and maternal ability traits in a multibreed sheep population from Brazil. AB - We determined the polymorphism C242T of the aromatase gene (Cyp19) and its allelic frequency, as well as the effect of the variants on productive and reproductive traits in 71 purebred Santa Ines sheep, 13 purebred Brazilian Somali sheep, nine purebred Poll Dorset sheep, and 18 crossbred 1/2 Dorper sheep. The animals were genotyped using the PCR-RFLP technique. The influence of the animal's genotype on its performance or on the performance of its lambs was analyzed by the least square method. Another factor assessed was the importance of the animal's genotype in analysis models for quantitative breeding value estimates, and whether there were differences among the averages of breeding values of animals with different genotypes for this gene. In the sample studied, no AA individuals were observed; the AB and BB frequencies were 0.64 and 0.36, respectively. All Brazilian Somali sheep were of genotype BB. All 1/2 Dorper BB animals presented a lower age at first lambing, and the Santa Ines BB ewes presented a lower lambing interval. In these same genetic groups, AB ewes presented higher litter weight at weaning. This is evidence that BB ewes have a better reproductive performance phenotype, whereas AB ewes present a better maternal ability phenotype. However, in general, animals with genotype AB presented better average breeding values than those with genotype BB. PMID- 21637511 TI - The association of CAPN1 316 marker genotypes with growth and meat quality traits of steers finished on pasture. AB - The objective of this paper was to determine the association of a SNP in the MU calpain gene at position 316 with growth and quality of meat traits of steers grown on pasture. Fifty-nine Brangus and 20 Angus steers were genotyped for CAPN1 316. Warner Bratzler shear force was measured in l. lumborum samples after a 7 day aging period. A multivariate analysis of variance was performed, including shear force (WBSF), final weight (FW), average daily gain (ADG), backfat thickness (BFT), average monthly fat thickness gain (AMFTG), rib-eye area (REA), and beef rib-eye depth (RED) as dependent variables. The CAPN1 316 genotype was statistically significant. Univariate analyses were done with these variables. The marker genotype was statistically significant (p < 0.05) for WBSF (kg: CC: 4.41 +/- 0.57; CG: 5.58 +/- 0.20; GG: 6.29 +/- 0.18), FW (kg: CC: 360.23 +/- 14.71; CG: 381.34 +/- 5.26; GG: 399.23 +/- 4.68), and ADG (kg/d: CC: 0.675 +/- 0.046; CG: 0.705 +/- 0.016; GG: 0.765 +/- 0.014) Shear force, final weight and average daily gain were significantly different according to the CAPN1 316 marker genotypes. The marker genotype was statistically significant in the multivariate analysis (p = 0.001). The first characteristic root explained 89% of the differences among genotypes. WBSF, FW and ADG were the most important traits in the first vector, indicating that animals with the marker genotype for lowest WBSF also have the lowest FW and ADG. PMID- 21637512 TI - Bias in the prediction of genetic gain due to mass and half-sib selection in random mating populations. AB - The prediction of gains from selection allows the comparison of breeding methods and selection strategies, although these estimates may be biased. The objective of this study was to investigate the extent of such bias in predicting genetic gain. For this, we simulated 10 cycles of a hypothetical breeding program that involved seven traits, three population classes, three experimental conditions and two breeding methods (mass and half-sib selection). Each combination of trait, population, heritability, method and cycle was repeated 10 times. The predicted gains were biased, even when the genetic parameters were estimated without error. Gain from selection in both genders is twice the gain from selection in a single gender only in the absence of dominance. The use of genotypic variance or broad sense heritability in the predictions represented an additional source of bias. Predictions based on additive variance and narrow sense heritability were equivalent, as were predictions based on genotypic variance and broad sense heritability. The predictions based on mass and family selection were suitable for comparing selection strategies, whereas those based on selection within progenies showed the largest bias and lower association with the realized gain. PMID- 21637513 TI - Inhibition of NF-kB 1 (NF-kBp50) by RNA interference in chicken macrophage HD11 cell line challenged with Salmonellaenteritidis. AB - The NF-kB pathway plays an important role in regulating the immunity response in animals. In this study, small interfering RNAs (siRNA) were used to specifically inhibit NF-kB 1 expression and to elucidate the role of NF-kB in the signal transduction pathway of the Salmonella challenge in the chicken HD11 cell line. The cells were transfected with either NF-kB 1 siRNA, glyceraldehyde 3-phosphate dehydrogenase siRNA (positive control) or the negative control siRNA for 24 h, followed by Salmonella enteritidis (SE) challenge or non-challenge for 1 h and 4 h. Eight candidate genes related to the signal pathway of SE challenge were selected to examine the effect of NF-kB 1 inhibition on their expressions by mRNA quantification. The results showed that, with a 36% inhibition of NF-kB 1 expression, gene expression of both Toll-like receptor (TLR) 4 and interleukin (IL)-6 was consistently and significantly increased at both 1 h and 4 h following SE challenge, whereas the gene expression of MyD88 and IL-1beta was increased at 1 h and 4 h, respectively. These findings suggest a likely inhibitory regulation by NF-kB 1, and could lay the foundation for studying the gene network of the innate immune response of SE infection in chickens. PMID- 21637514 TI - Regulation of MMP2 and MMP9 metalloproteinases by FSH and growth factors in bovine granulosa cells. AB - Matrix metalloproteinases (MMP) are key enzymes involved in tissue remodeling. Within the ovary, they are believed to play a major role in ovulation, and have been linked to follicle atresia. To gain insight into the regulation of MMPs, we measured the effect of hormones and growth factors on MMP2 and MMP9 mRNA levels in non-luteinizing granulosa cells in serum-free culture. FSH and IGF1 both stimulated estradiol secretion and inhibited MMP2 and MMP9 mRNA abundance. In contrast, EGF and FGF2 both inhibited estradiol secretion but had no effect on MMP expression. At physiological doses, none of these hormones altered the proportion of dead cells. Although we cannot link MMP expression with apoptosis, the specific down regulation by the gonadotropic hormones FSH and IGF1 in vitro suggests that excess MMP2 and MMP9 expression is neither required nor desired for follicle development. PMID- 21637515 TI - Genetic diversity and phylogeny in Hystrix (Poaceae, Triticeae) and related genera inferred from Giemsa C-banded karyotypes. AB - The phylogenetic relationships of 15 taxa from Hystrix and the related genera Leymus (NsXm), Elymus (StH), Pseudoroegneria (St), Hordeum (H), Psathyrostachys (Ns), and Thinopyrum (E) were examined by using the Giemsa C-banded karyotype. The Hy. patula C-banding pattern was similar to those of Elymus species, whereas C-banding patterns of the other Hystrix species were similar to those of Leymus species. The results suggest high genetic diversity within Hystrix, and support treating Hy. patula as E. hystrix L., and transferring Hy. coreana, Hy. duthiei ssp. duthiei and Hy. duthiei ssp. longearistata to the genus Leymus. On comparing C-banding patterns of Elymus species with their diploid ancestors (Pseudoroegneria and Hordeum), there are indications that certain chromosomal re arrangements had previously occurred in the St and H genomes. Furthermore, a comparison of the C-banding patterns of the Hystrix and Leymus species with the potential diploid progenitors (Psathyrostachys and Thinopyrum) suggests that Hy. coreana and some Leymus species are closely related to the Ns genome of Psathyrostachys, whereas Hy. duthiei ssp. duthiei, Hy. duthiei ssp. longearistata and some of the Leymus species have a close relationship with the E genome. The results suggest a multiple origin of the polyploid genera Hystrix and Leymus. PMID- 21637516 TI - Karyotype and nuclear DNA content of hexa-, octo-, and duodecaploid lines of Bromus subgen. Ceratochloa. AB - The subgenus Ceratochloa of the genus Bromus includes a number of closely related allopolyploid forms or species that present a difficult taxonomic problem. The present work combines data concerning chromosome length, heterochromatin distribution and nuclear genome size of different 6x, 8x and 12x accessions in this subgenus. Special attention is paid to the karyotype structure and genomic constitution of duodecaploid plants recently found in South America. Hexaploid lineages possess six almost indistinguishable genomes and a nuclear DNA content between 12.72 pg and 15.10 pg (mean 1Cx value = 2.32 pg), whereas octoploid lineages contain the same six genomes (AABBCC) plus two that are characterized by longer chromosomes and a greater DNA content (1Cx = 4.47 pg). Two duodecaploid accessions found in South America resemble each other and apparently differ from the North American duodecaploid B. arizonicus as regards chromosome size and nuclear DNA content (40.00 and 40.50 pg vs. 27.59 pg). These observations suggest that the South American duodecaploids represent a separate evolutionary lineage of the B. subgenus Ceratochloa, unrecognized heretofore. PMID- 21637517 TI - Development of a Brazilian maize core collection. AB - The aim of this study was to evaluate methods for developing a Brazilian maize core collection. For an initial survey of the active collection, passport information, as well as characterization and evaluation of accessions, were taken into consideration, these then being divided according to geographic region and kernel-type. Multiple sampling methods were evaluated. The strategy of constant sampling generated extensive alterations in extract accession frequency. The multivariate strategy with dispersion graphs and principal components associated with the Tocher method was considered efficient for identifying the most divergent genotypes. The multivariate strategy generated greater alterations in the variance of traits. The average number of traits revealed few modifications with the various sampling strategies used. Therefore, the active collection could be considered as possessing a satisfactory amount of information for most of its accessions. Moreover, the multivariate strategy generated modifications in the variance of the traits, independent of sampling intensity. PMID- 21637518 TI - Patterns of genetic diversity in southern and southeastern Araucaria angustifolia (Bert.) O. Kuntze relict populations. AB - Habitat fragmentation and a decrease in population size may lead to a loss in population genetic diversity. For the first time, the reduction in genetic diversity in the northernmost limit of natural occurence (southeastern Brazil) of Araucaria angustifolia in comparison with populations in the main area of the species continuous natural distribution (southern Brazil), was tested. The 673 AFLPs markers revealed a high level of genetic diversity for the species (Ht = 0.27), despite anthropogenic influence throughout the last century, and a decrease of H in isolated populations of southeastern Brazil (H = 0.16), thereby indicating the tendency for higher genetic diversity in remnant populations of continuous forests in southern Brazil, when compared to natural isolated populations in the southeastern region. A strong differentiation among southern and southeastern populations was detected (AMOVA variance ranged from 10%-15%). From Bayesian analysis, it is suggested that the nine populations tested form five "genetic clusters" (K = 5). Five of these populations, located in the northernmost limit of distribution of the species, represent three "genetic clusters". These results are in agreement with the pattern of geographic distribution of the studied populations. PMID- 21637519 TI - Genetic variability in Brazilian wheat cultivars assessed by microsatellite markers. AB - Wheat (Triticum aestivum) is one of the most important food staples in the south of Brazil. Understanding genetic variability among the assortment of Brazilian wheat is important for breeding. The aim of this work was to molecularly characterize the thirty-six wheat cultivars recommended for various regions of Brazil, and to assess mutual genetic distances, through the use of microsatellite markers. Twenty three polymorphic microsatellite markers (PMM) delineated all 36 of the samples, revealing a total of 74 simple sequence repeat (SSR) alleles, i.e. an average of 3.2 alleles per locus. Polymorphic information content (PIC value) calculated to assess the informativeness of each marker ranged from 0.20 to 0.79, with a mean of 0.49. Genetic distances among the 36 cultivars ranged from 0.10 (between cultivars Ocepar 18 and BRS 207) to 0.88 (between cultivars CD 101 and Fudancep 46), the mean distance being 0.48. Twelve groups were obtained by using the unweighted pair-group method with arithmetic means analysis (UPGMA), and thirteen through the Tocher method. Both methods produced similar clusters, with one to thirteen cultivars per group. The results indicate that these tools may be used to protect intellectual property and for breeding and selection programs. PMID- 21637520 TI - Conversion of barley SNPs into PCR-based markers using dCAPS method. AB - Molecular genetic research relies heavily on the ability to detect polymorphisms in DNA. Single nucleotide polymorphisms (SNPs) are the most frequent form of DNA variation in the genome. In combination with a PCR assay, the corresponding SNP can be analyzed as a derived cleaved amplified polymorphic sequence (dCAPS) marker. The dCAPS method exploits the well-known specificity of a restriction endonuclease for its recognition site and can be used to virtually detect any SNP. Here, we describe the use of the dCAPS method for detecting single nucleotide changes by means of a barley EST, CK569932, PCR-based marker. PMID- 21637521 TI - Transferability and characterization of microsatellite markers in two Neotropical Ficus species. AB - Microsatellite markers were transferred and characterized for two Neotropical fig tree species, Ficus citrifolia and Ficus eximia. Our study demonstrated that microsatellite markers developed from different subgenera of Ficus can be transferred to related species. In the present case, 12 of the 15 primer pairs tested (80%) were successfully transferred to both of the above species. Eleven loci were polymorphic when tested across 60 F. citrifolia and 60 F. eximia individuals. For F. citrifolia, there were 4 to 15 alleles per locus, whereas expected heterozygosities ranged from 0.31 to 0.91. In the case of F. eximia, this was 2 to 12 alleles per locus and expected heterozygosities from 0.42 to 0.87. PMID- 21637522 TI - Physiological traits of the symbiotic bacterium Teredinibacter turnerae isolated from the mangrove shipworm Neoteredo reynei. AB - Nutrition in the Teredinidae family of wood-boring mollusks is sustained by cellulolytic/nitrogen fixing symbiotic bacteria of the Teredinibacter clade. The mangrove Teredinidae Neoteredo reynei is popularly used in the treatment of infectious diseases in the north of Brazil. In the present work, the symbionts of N. reynei, which are strictly confined to the host's gills, were conclusively identified as Teredinibacter turnerae. Symbiont variants obtained in vitro were able to grow using casein as the sole carbon/nitrogen source and under reduced concentrations of NaCl. Furthermore, cellulose consumption in T. turnerae was clearly reduced under low salt concentrations. As a point of interest, we hereby report first hand that T. turnerae in fact exerts antibiotic activity. Furthermore, this activity was also affected by NaCl concentration. Finally, T. turnerae was able to inhibit the growth of Gram-negative and Gram-positive bacteria, this including strains of Sphingomonas sp., Stenotrophomonas maltophilia, Bacillus cereus and Staphylococcus sciuri. Our findings introduce new points of view on the ecology of T. turnerae, and suggest new biotechnological applications for this marine bacterium. PMID- 21637523 TI - Sequence and expression pattern of the germ line marker vasa in honey bees and stingless bees. AB - Queens and workers of social insects differ in the rates of egg laying. Using genomic information we determined the sequence of vasa, a highly conserved gene specific to the germ line of metazoans, for the honey bee and four stingless bees. The vasa sequence of social bees differed from that of other insects in two motifs. By RT-PCR we confirmed the germ line specificity of Amvasa expression in honey bees. In situ hybridization on ovarioles showed that Amvasa is expressed throughout the germarium, except for the transition zone beneath the terminal filament. A diffuse vasa signal was also seen in terminal filaments suggesting the presence of germ line cells. Oocytes showed elevated levels of Amvasa transcripts in the lower germarium and after follicles became segregated. In previtellogenic follicles, Amvasa transcription was detected in the trophocytes, which appear to supply its mRNA to the growing oocyte. A similar picture was obtained for ovarioles of the stingless bee Melipona quadrifasciata, except that Amvasa expression was higher in the oocytes of previtellogenic follicles. The social bees differ in this respect from Drosophila, the model system for insect oogenesis, suggesting that changes in the sequence and expression pattern of vasa may have occurred during social evolution. PMID- 21637524 TI - Molecular cloning and expression analysis of a zebrafish novel zinc finger protein gene rnf141. AB - ZNF230 is a novel zinc finger gene cloned by our laboratory. In order to understand the potential functions of this gene in vertebrate development, we cloned the zebrafish orthologue of human ZNF230, named rnf141. The cDNA fragment of rnf141 was obtained by rapid amplification of cDNA ends (RACE). The open reading frame (ORF) encodes a polypeptide of 222 amino acids which shares 75.65% identity with the human ZNF230. RT-PCR analysis in zebrafish embryo and adult tissues revealed that rnf141 transcripts are maternally derived and that rnf141 mRNA has a broad distribution. Zygotic rnf141 message is strongly localized in the central nervous system, as shown by whole-mount in situ hybridization. Knockdown and over expression of rnf141 can induce abnormal phenotypes, including abnormal development of brain, as well as yolk sac and axis extendsion. Marker gene analysis showed that rnf141 may play a role in normal dorsoventral patterning of zebrafish embryos, suggesting that rnf141 may have a broad function during early development of vertebrates. PMID- 21637525 TI - Genetic characterization of native and introduced populations of the neotropical cichlid genus Cichla in Brazil. AB - A molecular phylogenetic analysis based on mitochondrial 16S ribosomal DNA and Control Region sequences from native and introduced populations was undertaken, in order to characterize the introduction of Cichla (peacock bass or tucunare) species in Brazil. Mitochondrial DNA haplotypes found in introduced fish from Minas Gerais state (southeastern Brazil) clustered only with those from native species of the Tocantins River (Cichla piquiti and C. kelberi), thereby suggesting a single or, at most, few translocation acts in this area, even though with fish from the same source-population. Our study contributes to an understanding of the introduction of Cichla in regions of Brazil outside the Amazon basin, and adds phylogenetic data to the recently describe Cichla species, endemic from the Tocantins-Araguaia basin. PMID- 21637526 TI - DNA barcoding of Brazilian sea turtles (Testudines). AB - Five out of the seven recognized species of sea turtles (Testudines) occur on the Brazilian coast. The Barcode Initiative is an effort to undertake a molecular inventory of Earth biodiversity. Cytochrome Oxidase c subunit I (COI) molecular tags for sea turtle species have not yet been described. In this study, COI sequences for the five species of sea turtles that occur in Brazil were generated. These presented widely divergent haplotypes. All observed values were on the same range as those already described for other animal groups: the overall mean distance was 8.2%, the mean distance between families (Dermochelyidae and Cheloniidae) 11.7%, the mean intraspecific divergence 0.34%, and the mean distance within Cheloniidae 6.4%, this being 19-fold higher than the mean divergence observed within species. We obtained species-specific COI barcode tags that can be used for identifying each of the marine turtle species studied. PMID- 21637527 TI - Green turtles (Chelonia mydas) foraging at Arvoredo Island in Southern Brazil: Genetic characterization and mixed stock analysis through mtDNA control region haplotypes. AB - We analyzed mtDNA control region sequences of green turtles (Chelonia mydas) from Arvoredo Island, a foraging ground in southern Brazil, and identified eight haplotypes. Of these, CM-A8 (64%) and CM-A5 (22%) were dominant, the remainder presenting low frequencies (< 5%). Haplotype (h) and nucleotide (pi) diversities were 0.5570 +/- 0.0697 and 0.0021 +/- 0.0016, respectively. Exact tests of differentiation and AMOVA Phi(ST) pairwise values between the study area and eight other Atlantic foraging grounds revealed significant differences in most areas, except Ubatuba and Rocas/Noronha, in Brazil (p > 0.05). Mixed Stock Analysis, incorporating eleven Atlantic and one Mediterranean rookery as possible sources of individuals, indicated Ascension and Aves islands as the main contributing stocks to the Arvoredo aggregation (68.01% and 22.96%, respectively). These results demonstrate the extensive relationships between Arvoredo Island and other Atlantic foraging and breeding areas. Such an understanding provides a framework for establishing adequate management and conservation strategies for this endangered species. PMID- 21637528 TI - A straightforward multiallelic significance test for the Hardy-Weinberg equilibrium law. AB - Much forensic inference based upon DNA evidence is made assuming Hardy-Weinberg Equilibrium (HWE) for the genetic loci being used. Several statistical tests to detect and measure deviation from HWE have been devised, and their limitations become more obvious when testing for deviation within multiallelic DNA loci. The most popular methods-Chi-square and Likelihood-ratio tests-are based on asymptotic results and cannot guarantee a good performance in the presence of low frequency genotypes. Since the parameter space dimension increases at a quadratic rate on the number of alleles, some authors suggest applying sequential methods, where the multiallelic case is reformulated as a sequence of "biallelic" tests. However, in this approach it is not obvious how to assess the general evidence of the original hypothesis; nor is it clear how to establish the significance level for its acceptance/rejection. In this work, we introduce a straightforward method for the multiallelic HWE test, which overcomes the aforementioned issues of sequential methods. The core theory for the proposed method is given by the Full Bayesian Significance Test (FBST), an intuitive Bayesian approach which does not assign positive probabilities to zero measure sets when testing sharp hypotheses. We compare FBST performance to Chi-square, Likelihood-ratio and Markov chain tests, in three numerical experiments. The results suggest that FBST is a robust and high performance method for the HWE test, even in the presence of several alleles and small sample sizes. PMID- 21637529 TI - Prediction of binding hot spot residues by using structural and evolutionary parameters. AB - In this work, we present a method for predicting hot spot residues by using a set of structural and evolutionary parameters. Unlike previous studies, we use a set of parameters which do not depend on the structure of the protein in complex, so that the predictor can also be used when the interface region is unknown. Despite the fact that no information concerning proteins in complex is used for prediction, the application of the method to a compiled dataset described in the literature achieved a performance of 60.4%, as measured by F-Measure, corresponding to a recall of 78.1% and a precision of 49.5%. This result is higher than those reported by previous studies using the same data set. PMID- 21637530 TI - D-MaPs - DNA-microarray projects: Web-based software for multi-platform microarray analysis. AB - The web application D-Maps provides a user-friendly interface to researchers performing studies based on microarrays. The program was developed to manage and process one- or two-color microarray data obtained from several platforms (currently, GeneTAC, ScanArray, CodeLink, NimbleGen and Affymetrix). Despite the availability of many algorithms and many software programs designed to perform microarray analysis on the internet, these usually require sophisticated knowledge of mathematics, statistics and computation. D-maps was developed to overcome the requirement of high performance computers or programming experience. D-Maps performs raw data processing, normalization and statistical analysis, allowing access to the analyzed data in text or graphical format. An original feature presented by D-Maps is GEO (Gene Expression Omnibus) submission format service. The D-MaPs application was already used for analysis of oligonucleotide microarrays and PCR-spotted arrays (one- and two-color, laser and light scanner). In conclusion, D-Maps is a valuable tool for microarray research community, especially in the case of groups without a bioinformatic core. PMID- 21637531 TI - Computational analysis suggests that virulence of Chromobacterium violaceum might be linked to biofilm formation and poly-NAG biosynthesis. AB - Groups of genes that produce exopolysaccharide with a N-acetyl-D-glucosamine monomer are in the genome of several pathogenic bacteria. Chromobacterium violaceum, an opportunistic pathogen, has the operon hmsHFR-CV2940, whose proteins can synthesize such polysaccharide. In this work, multiple alignments among proteins from bacteria that synthesize such polysaccharide were used to verify the existence of amino acids that might be critical for pathogen activity. Three-dimensional models were generated for spatial visualization of these amino acid residues. The analysis carried out showed that the protein HmsR preserves the amino acids D135, D228, Q264 and R267, considered critical for the formation of biofilms and, furthermore, that these amino acids are close to each other. The protein HmsF of C. violaceum preserves the residues D86, D87, H156 and W115. It was also shown that these residues are also close to each other in their spatial arrangement. For the proteins HmsH and CV2940 there is evidence of conservation of the residues R104 and W94, respectively. Conservation and favorable spatial location of those critical amino acids that constitute the proteins of the operon indicates that they preserve the same enzymatic function in biofilm synthesis. This is an indicator that the operon hmsHFR-CV2940 is a possible target in C. violaceum pathogenicity. PMID- 21637532 TI - Using linear algebra for protein structural comparison and classification. AB - In this article, we describe a novel methodology to extract semantic characteristics from protein structures using linear algebra in order to compose structural signature vectors which may be used efficiently to compare and classify protein structures into fold families. These signatures are built from the pattern of hydrophobic intrachain interactions using Singular Value Decomposition (SVD) and Latent Semantic Indexing (LSI) techniques. Considering proteins as documents and contacts as terms, we have built a retrieval system which is able to find conserved contacts in samples of myoglobin fold family and to retrieve these proteins among proteins of varied folds with precision of up to 80%. The classifier is a web tool available at our laboratory website. Users can search for similar chains from a specific PDB, view and compare their contact maps and browse their structures using a JMol plug-in. PMID- 21637533 TI - Actin-interacting and flagellar proteins in Leishmania spp.: Bioinformatics predictions to functional assignments in phagosome formation. AB - Several motile processes are responsible for the movement of proteins into and within the flagellar membrane, but little is known about the process by which specific proteins (either actin-associated or not) are targeted to protozoan flagellar membranes. Actin is a major cytoskeleton protein, while polymerization and depolymerization of parasite actin and actin-interacting proteins (AIPs) during both processes of motility and host cell entry might be key events for successful infection. For a better understanding the eukaryotic flagellar dynamics, we have surveyed genomes, transcriptomes and proteomes of pathogenic Leishmania spp. to identify pertinent genes/proteins and to build in silico models to properly address their putative roles in trypanosomatid virulence. In a search for AIPs involved in flagellar activities, we applied computational biology and proteomic tools to infer from the biological meaning of coronins and Arp2/3, two important elements in phagosome formation after parasite phagocytosis by macrophages. Results presented here provide the first report of Leishmania coronin and Arp2/3 as flagellar proteins that also might be involved in phagosome formation through actin polymerization within the flagellar environment. This is an issue worthy of further in vitro examination that remains now as a direct, positive bioinformatics-derived inference to be presented. PMID- 21637534 TI - Interleukin-8-251T > A, Interleukin-1alpha-889C > T and Apolipoprotein E polymorphisms in Alzheimer's disease. AB - An inflammatory process has been involved in numerous neurodegenerative disorders such as Parkinson's disease, stroke and Alzheimer's disease (AD). In AD, the inflammatory response is mainly located in the vicinity of amyloid plaques. Cytokines, such as interleukin-8 (IL-8) and interleukin-1alpha (IL-1alpha), have been clearly involved in this inflammatory process. Polymorphisms of several interleukin genes have been correlated to the risk of developing AD. The present study investigated the association of AD with polymorphisms IL-8 -251T > A (rs4073) and IL-1alpha-889C > T (rs1800587) and the interactive effect of both, adjusted by the Apolipoprotein E genotype. 199 blood samples from patients with AD, 146 healthy elderly controls and 95 healthy young controls were obtained. DNA samples were isolated from blood cells, and the PCR-RFLP method was used for genotyping. The genotype distributions of polymorphisms IL-8, IL-1alpha and APOE were as expected under Hardy-Weinberg equilibrium. The allele frequencies did not differ significantly among the three groups tested. As expected, the APOE4 allele was strongly associated with AD (p < 0.001). No association of AD with either the IL-1alpha or the IL-8 polymorphism was observed, nor was any interactive effect between both polymorphisms. These results confirm previous studies in other populations, in which polymorphisms IL-8 -251T > A and IL-1alpha-889C > T were not found to be risk factors for AD. PMID- 21637535 TI - Transcriptional activity of the 5'-flanking region of the thyroid transcription factor-1 gene in human thyroid cell lines. AB - Thyroid transcription factor-1 (TTF-1, NKX2-1) is a homeodomain-containing transcriptional factor that binds to and activates the promoters of thyroid and lung-specific genes, such as thyroglobulin, thyroid peroxidase, and thyroid stimulating hormone receptor. TTF-1 is known to play a key role in the development of the thyroid. However, the precise mechanism of TTF-1 gene transcription in human thyroid cells has not been studied. The expression of transcriptional activity in various lengths of the 5'-flanking region of the human TTF -1 gene was studied in TTF-1 positive and negative human thyroid cell lines. Increased transcriptional activity was observed in thyroid cell lines containing plasmids that coded for a sequence proximal to the transcription start site of exon 1 of the TTF-1 gene. However, we did not observe any difference in promoter activity in the region up to -2.6 kb from the proximal transcription start site of the TTF-1 gene between TTF-1 positive and negative cells. These results suggest that the proximal 5'-flanking region of the human TTF -1 gene does not contain sufficient cis-active regulatory information to direct gene expression in thyroid cells, and that other cis- or trans-acting factors participate in the thyroid specific gene expression of TTF-1. PMID- 21637536 TI - Superoxide dismutase, catalase, glutathione peroxidase and gluthatione S transferases M1 and T1 gene polymorphisms in three Brazilian population groups. AB - Antioxidants such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX1) reduce the oxidation rates in the organism. Gluthatione S transferases (GSTs) play a vital role in phase 2 of biotransformation of many substances. Variation in the expression of these enzymes suggests individual differences for the degree of antioxidant protection and geographical differences in the distribution of these variants. We described the distribution frequency of CAT (21A/T), SOD2 (Ala9Val), GPX1 (Pro198Leu), GSTM1 and GSTT1 polymorphisms in three Brazilian population groups: Kayabi Amerindians (n = 60), Kalunga Afro descendants (n = 72), and an urban mixed population from Federal District (n = 162). Frequencies of the variants observed in Kalunga (18% to 58%) and Federal District (33% to 63%) were similar to those observed in Euro and Afro descendants, while in Kayabi (3% to 68%), depending on the marker, frequencies were similar to the ones found in different ethnic groups. Except for SOD2 in all population groups studied here, and for GPX1 in Kalunga, the genotypic distributions were in accordance with Hardy-Weinberg Equilibrium. These data can clarify the contribution of different ethnicities in the formation of mixed populations, such as that of Brazil. Moreover, outcomes will be valuable resources for future functional studies and for genetic studies in specific populations. If these studies are designed to comprehensively explore the role of these genetic polymorphisms in the etiology of human diseases they may help to prevent inconsistent genotype-phenotype associations in pharmacogenetic studies. PMID- 21637537 TI - Regulation of adipogenesis by nuclear receptor PPARgamma is modulated by the histone demethylase JMJD2C. AB - A potential strategy to combat obesity and its associated complications involves modifying gene expression in adipose cells to reduce lipid accumulation. The nuclear receptor Peroxisome Proliferator-activated receptor gamma (PPARgamma) is the master regulator of adipose cell differentiation and its functional activation is currently used as a therapeutic approach for Diabetes Mellitus type 2. However, total activation of PPARgamma induces undesirable secondary effects that might be set with a partial activation. A group of proteins that produce histone demethylation has been shown to modify the transcriptional activity of nuclear receptors. Here we describe the repressive action of the jumonji domain containing 2C/lysine demethylase 4 C (JMJD2C/KDM4C) on PPARgamma transcriptional activation. JMJD2C significantly reduced the rosiglitazone stimulated PPARgamma activation. This effect was mainly observed in experiments performed using the Tudor domains that may interact with histone deacetylase class 1 (HDAC) and this interaction probably reduces the mediated activation of PPARgamma. Trichostatin A, a HDAC inhibitor, reduces the repressive effect of JMJD2C. When JMJD2C was over-expressed in 3T3-L1 cells, a reduction of differentiation was observed with the Tudor domain. In summary, we herein describe JMJD2C-mediated reduction of PPARgamma transcriptional activation as well as preadipocyte differentiation. This novel action of JMJD2C might have an important role in new therapeutic approaches to treat obesity and its complications. PMID- 21637538 TI - Impact of the tissue factor pathway inhibitor gene on apoptosis in human vascular smooth muscle cells. AB - Tissue factor pathway inhibitor (TFPI) plays a vitally important role in the blood coagulation pathway. Recent studies indicated that TFPI induces apoptosis in vascular smooth-muscle cells (VSMCs) in animals. The present study investigated whether the TFPI gene could also induce apoptosis in human vascular smooth-muscle cells (hVSMCs). Such cells were isolated from human umbilical arteries and subsequently transfected with pIRES-TFPI plasmid (2 MUg/mL). MTT assaying and cell counting were applied to measure cell viability and proliferation, RT-PCR was utilized to analyze TFPI gene expression in the cells. Apoptosis was analyzed by fluorescence activated cell sorting (FACS). Several key proteins involved in apoptosis were examined through Western blotting. It was shown that TFPI gene transfer led to its increased cellular expression, with a subsequent reduction in hVSMC proliferation. Further investigation demonstrated that TFPI gene expression resulted in lesser amounts of procaspase-3, procaspase 8 and procascase-9, and an increased release of mitochondrial cytochrome c (cyt c) into cytoplasm, thereby implying the involvement of both extrinsic and intrinsic pathways in TFPI gene-induced apoptosis in hVSMCs. PMID- 21637539 TI - Human aging and somatic point mutations in mtDNA: A comparative study of generational differences (grandparents and grandchildren). AB - The accumulation of somatic mutations in mtDNA is correlated with aging. In this work, we sought to identify somatic mutations in the HVS-1 region (D-loop) of mtDNA that might be associated with aging. For this, we compared 31 grandmothers (mean age: 63 +/- 2.3 years) and their 62 grandchildren (mean age: 15 +/- 4.1 years), the offspring of their daughters. Direct DNA sequencing showed that mutations absent in the grandchildren were detected in a presumably homoplasmic state in three grandmothers and in a heteroplasmic state in an additional 13 grandmothers; no mutations were detected in the remaining 15 grandmothers. However, cloning followed by DNA sequencing in 12 grandmothers confirmed homoplasia in only one of the three mutations previously considered to be homoplasmic and did not confirm heteroplasmy in three out of nine grandmothers found to be heteroplasmic by direct sequencing. Thus, of 12 grandmothers in whom mtDNA was analyzed by cloning, eight were heteroplasmic for mutations not detected in their grandchildren. In this study, the use of genetically related subjects allowed us to demonstrate the occurrence of age-related (> 60 years old) mutations (homoplasia and heteroplasmy). It is possible that both of these situations (homoplasia and heteroplasmy) were a long-term consequence of mitochondrial oxidative phosphorylation that can lead to the accumulation of mtDNA mutations throughout life. PMID- 21637540 TI - Allelic frequencies and statistical data obtained from 12 codis STR loci in an admixed population of the Brazilian Amazon. AB - The allelic frequencies of 12 short tandem repeat loci were obtained from a sample of 307 unrelated individuals living in Macapa, a city in the northern Amazon region, Brazil. These loci are the most commonly used in forensics and paternity testing. Based on the allele frequency obtained for the population of Macapa, we estimated an interethnic admixture for the three parental groups (European, Native American and African) of, respectively, 46%, 35% and 19%. Comparing these allele frequencies with those of other Brazilian populations and of the Iberian Peninsula population, no significant distances were observed. The interpopulation genetic distances (F(ST) coefficients) to the present database ranged from F(ST) = 0.0016 between Macapa and Belem to F(ST) = 0.0036 between Macapa and the Iberian Peninsula. PMID- 21637541 TI - Biochemical and genetic analysis of butyrylcholinesterase (BChE) in a family, due to prolonged neuromuscular blockade after the use of succinylcholine. AB - Butyrylcholinesterase (BChE) is a plasma enzyme that catalyzes the hydrolysis of choline esters, including the muscle-relaxant succinylcholine and mivacurium. Patients who present sustained neuromuscular blockade after using succinylcholine usually carry BChE variants with reduced enzyme activity or an acquired BChE deficiency. We report here the molecular basis of the BCHE gene underlying the slow catabolism of succinylcholine in a patient who underwent endoscopic nasal surgery. We measured the enzyme activity of BChE and extracted genomic DNA in order to study the promoter region and all exons of the BCHE gene of the patient, her parents and siblings. PCR products were sequenced and compared with reference sequences from GenBank. We detected that the patient and one of her brothers have two homozygous mutations: nt1615 GCA > ACA (Ala539Thr), responsible for the K variant, and nt209 GAT > GGT (Asp70Gly), which produces the atypical variant A. Her parents and two of her brothers were found to be heterozygous for the AK allele, and another brother is homozygous for the normal allele. Sequence analysis of exon 1 including 5'UTR showed that the proband and her brother are homozygous for -116GG. The AK/AK genotype is considered the most frequent in hereditary hypocholinesterasemia (44%). This work demonstrates the importance of defining the phenotype and genotype of the BCHE gene in patients who are subjected to neuromuscular block by succinylcholine, because of the risk of prolonged neuromuscular paralysis. PMID- 21637542 TI - Population analysis of the GLB1 gene in South Brazil. AB - Infantile GM1 gangliosidosis is caused by the absence or reduction of lysosomal beta-galactosidase activity. Studies conducted in Brazil have indicated that it is one of the most frequent lysosomal storage disorders in the southern part of the country. To assess the incidence of this disorder, 390 blood donors were tested for the presence of two common mutations (1622-1627insG and R59H) in the GLB1 gene. Another group, consisting of 26 GM1 patients, and the blood donors were tested for the presence of two polymorphisms (R521C and S532G), in an attempt to elucidate whether there is a founder effect. The frequencies of the R59H and 1622-1627insG mutations among the GM1 patients studied were 19.2% and 38.5%, respectively. The frequency of polymorphism S532G was 16.7%, whereas R521C was not found in the patients. The overall frequency of either R59H or 1622 1627insG was 57.7% of the disease-causing alleles. This epidemiological study suggested a carrier frequency of 1:58. Seven different haplotypes were found. The 1622-1627insG mutation was not found to be linked to any polymorphism, whereas linkage disequilibrium was found for haplotype 2 (R59H, S532G) (p < 0.001). These data confirm the high incidence of GM1 gangliosidosis and the high frequency of two common mutations in southern Brazil. PMID- 21637543 TI - The association of two single nucleotide polymorphisms (SNPs) in growth hormone (GH) gene with litter size and superovulation response in goat-breeds. AB - Two active mutations (A 781 G and A 1575 G) in growth hormone (GH) gene, and their associations with litter size (LS), were investigated in both a high prolificacy (Matou, n = 182) and a low prolificacy breed (Boer, n = 352) by using the PCR-RFLP method. Superovulation experiments were designed in 57 dams, in order to evaluate the effect of different genotypes of the GH gene on superovulation response. Two genotypes (AA and AB, CC and CD) in each mutation were detected in these two goat breeds. Neither BB nor DD homozygous genotypes were observed. The genotypic frequencies of AB and CC were significantly higher than those of AA and CD. In the third parity, Matou dams with AB or CC genotypes had significantly larger litter sizes than those with AA and CD (p < 0.05). On combining the two loci, both Matou and Boer dams with ABCD genotype had the largest litter sizes when compared to the other genotypes (p < 0.05). When undergoing like superovulation treatments, a significantly higher number of corpora lutea and ova, with a lower incidence of ovarian cysts, were harvested in the AB and CC genotypes than in AA and CD. These results show that the two loci of GH gene are highly associated with abundant prolificacy and superovulation response in goat breeds. PMID- 21637544 TI - Association of PPARgamma2 polymorphisms with carcass and meat quality traits in a Pietrain x Jinhua F2 population. AB - The PPARgamma2 gene is a key regulator of both proliferation and preadipocyte differentiation in mammals. Herein its genotype and allele frequencies were analyzed using PCR-SSCP in eight pig breeds (N = 416). Two kinds of polymorphisms of the PPARgamma2 gene were detected, including a previously reported shift SNP A177G (Met59Val) in exon 1 and a novel silent mutation G876A in exon 5. The results revealed that European pig breeds carry a higher allele A frequency at the A177G locus and a fixed GG genotype at the G876A locus. Allele A at the G876A locus was only found in Jinhua pigs. The association between haplotype (A177G/G876A) and carcass and meat quality traits was analyzed in a Pietrain x Jinhua F2 population (N = 248). The PPARgamma2 gene was found to be significantly associated with backfat thickness at the shoulder (p < 0.05), 6-7(th) ribs (p < 0.01), last rib (p < 0.01), gluteus medius (p <0.05) and ham weight (p < 0.01). Significant effects of different haplotypes on ham weight and backfat thickness at the 6-7(th) ribs, last rib, and gluteus medius were also observed. PMID- 21637545 TI - Effect of lactation length adjustment procedures on genetic parameter estimates for buffalo milk yield. AB - The objectives of this study were to estimate the genetic parameters for milk yield unadjusted and adjusted for days in milk and, subsequently, to assess the influence of adjusting for days in milk on sire rank. Complete lactations from 90 or 150 days of lactation to 270 or 350 days in milk were considered in these analyses. Milk yield was adjusted for days in milk by multiplicative correction factors, or by including lactation length as a covariable in the model. Milk yields adjusted by different procedures were considered as different traits. Heritability estimates varied from 0.17 to 0.28. Genetic correlation estimates between milk yields unadjusted and adjusted for days in milk were greater than 0.82. Adjusting for days in milk affected the parameter estimates. Multiplicative correction factors produced the highest heritability estimates. More reliable breeding value estimates can be expected by including short length lactation records in the analyses and adjusting the milk yields for days in milk, regardless of the method used for the adjustment. High selection intensity coupled to the inclusion of short length lactations and adjustment with multiplicative factors can change the sire rank.. PMID- 21637546 TI - Genetic variability in the Skyros pony and its relationship with other Greek and foreign horse breeds. AB - In Greece, seven native horse breeds have been identified so far. Among these, the Skyros pony is outstanding through having a distinct phenotype. In the present study, the aim was to assess genetic diversity in this breed, by using different types of genetic loci and available genealogical information. Its relationships with the other Greek, as well as foreign, domestic breeds were also investigated. Through microsatellite and pedigree analysis it appeared that the Skyros presented a similar level of genetic diversity to the other European breeds. Nevertheless, comparisons between DNA-based and pedigree-based results revealed that a loss of genetic diversity had probably already occurred before the beginning of breed registration. Tests indicated the possible existence of a recent bottleneck in two of the three main herds of Skyros pony. Nonetheless, relatively high levels of heterozygosity and Polymorphism Information Content indicated sufficient residual genetic variability, probably useful in planning future strategies for breed conservation. Three other Greek breeds were also analyzed. A comparison of these with domestic breeds elsewhere, revealed the closest relationships to be with the Middle Eastern types, whereas the Skyros itself remained isolated, without any close relationship, whatsoever. PMID- 21637547 TI - Occurrence of B chromosomes in Tetragonisca Latreille, 1811 (Hymenoptera, Apidae, Meliponini): A new contribution to the cytotaxonomy of the genus. AB - Tetragonisca angustula and Tetragonisca fiebrigi have recently been listed as valid species. This study aimed to cytogenetically investigate both species, emphasizing the new registry of B chromosomes in the tribe Meliponini. We analyzed colonies of T. angustula and T. fiebrigi collected at Tangara da Serra, Mato Grosso, Brazil, through conventional Giemsa staining, C-banding, and base specific fluorochrome staining (CMA(3)/DAPI). T. angustula showed 2n = 34 chromosomes in females and n = 17 in males, with karyotype formula 2K = 34A(M). T. fiebrigi showed numeric variation, with chromosome number varying from 2n = 34 to 2n = 36 in females and from n = 17 to n = 18 in males, with karyotype formula 2K = 32A(M)+2A(Mc) and 2K = 32A(M)+2A(Mc) + 1 or 2 B-chromosomes. The B chromosomes are heterochromatic. In T. fiebrigi, the CMA(3)/DAPI staining revealed four chromosomes with a CMA(3) positive band. All individuals from the same colony showed the same number of B chromosomes. T. angustula and T. fiebrigi showed karyotype divergence, principally due to the presence of B chromosomes, which are found only in T. fiebrigi. Our data corroborate the status of valid species for both T. angustula and T. fiebrigi, as recently proposed. PMID- 21637548 TI - Maintenance of syntenic groups between Cathartidae and Gallus gallus indicates symplesiomorphic karyotypes in new world vultures. AB - Similarities between New World and Old World vultures have been interpreted to reflect a close relationship and to suggest the inclusion of both in Accipitridae (Falconiformes). However, deeper analyses indicated that the placement of the New World vultures (cathartids) in this Order is uncertain. Chromosome analysis has shown that cathartids retained a karyotype similar to the putative avian ancestor. In order to verify the occurrence of intrachromosomal rearrangements in cathartids, we hybridized whole chromosome probes of two species (Gallus gallus and Leucopternis albicollis) onto metaphases of Cathartes aura. The results showed that not only were the syntenic groups conserved between Gallus and C. aura, but probably also the general gene order, suggesting that New World vultures share chromosomal symplesiomorphies with most bird lineages. PMID- 21637549 TI - The karyotype of Adenomera diptyx (Boettger 1885) (Anura, Leptodactylidae) from northeastern Argentina. AB - In this work we analyzed the karyotype of five populations of Adenomera diptyx from Argentina after conventional staining, Ag-NOR and C-banding. All specimens presented 2n = 26 and FN = 34. The karyotype was formed by three submetacentric, one metacentric and nine telocentric pairs. Silver staining revealed that the NOR was located on a secondary constriction in pair 7. C- banding evidenced constitutive heterochromatin at the pericentromeric region of all chromosomes. The karyotype of A. diptyx was similar to that of A. hylaedactyla (2n = 26, FN = 34) and different from that of A. andreae (2n = 26, FN = 40) in the fundamental number and secondary constriction position. It also differed from the karyotypes of A. marmorata (2n = 24, FN = 34 and 36) and of A. aff. bokermanni (2n = 23, FN = 34) in diploid number. Until a comprehensive cytogenetic analysis of all the species of the genus is performed, their chromosome evolution will remain poorly understood. PMID- 21637550 TI - Genetic diversity in cultivated carioca common beans based on molecular marker analysis. AB - A wide array of molecular markers has been used to investigate the genetic diversity among common bean species. However, the best combination of markers for studying such diversity among common bean cultivars has yet to be determined. Few reports have examined the genetic diversity of the carioca bean, commercially one of the most important common beans in Brazil. In this study, we examined the usefulness of two molecular marker systems (simple sequence repeats - SSRs and amplified fragment length polymorphisms - AFLPs) for assessing the genetic diversity of carioca beans. The amount of information provided by Roger's modified genetic distance was used to analyze SSR data and Jaccards similarity coefficient was used for AFLP data. Seventy SSRs were polymorphic and 20 AFLP primer combinations produced 635 polymorphic bands. Molecular analysis showed that carioca genotypes were quite diverse. AFLPs revealed greater genetic differentiation and variation within the carioca genotypes (Gst = 98% and Fst = 0.83, respectively) than SSRs and provided better resolution for clustering the carioca genotypes. SSRs and AFLPs were both suitable for assessing the genetic diversity of Brazilian carioca genotypes since the number of markers used in each system provided a low coefficient of variation. However, fingerprint profiles were generated faster with AFLPs, making them a better choice for assessing genetic diversity in the carioca germplasm. PMID- 21637551 TI - Characterization of a small cryptic plasmid from endophytic Pantoea agglomerans and its use in the construction of an expression vector. AB - A circular cryptic plasmid named pPAGA (2,734 bp) was isolated from Pantoea agglomerans strain EGE6 (an endophytic bacterial isolate from eucalyptus). Sequence analysis revealed that the plasmid has a G+C content of 51% and contains four potential ORFs, 238(A), 250(B), 131(C), and 129(D) amino acids in length without homology to known proteins. The shuttle vector pLGM1 was constructed by combining the pPAGA plasmid with pGFPmut3.0 (which harbors a gene encoding green fluorescent protein, GFP), and the resulting construct was used to over-express GFP in E. coli and P. agglomerans cells. GFP production was used to monitor the colonization of strain EGE6gfp in various plant tissues by fluorescence microscopy. Analysis of EGE6gfp colonization showed that 14 days after inoculation, the strain occupied the inner tissue of Eucalyptus grandis roots, preferentially colonizing the xylem vessels of the host plants. PMID- 21637552 TI - Induction of SCEs and DNA fragmentation in bovine peripheral lymphocytes by in vitro exposure to tolylfluanid-based fungicide. AB - The potential for genotoxic and cytotoxic effects of tolylfluanid-based fungicide (50% active agent) was evaluated using sister chromatid exchange (SCE) and proliferation indices (PI) in cultured bovine peripheral lymphocytes. For the detection of possible genetic damage, DNA fragmentation assay was also applied. Bovine lymphocytes cultured for 72 h were treated with the fungicide at the final concentrations of 1.75, 3.5, 8.75, and 17.5 MUg/mL for the last 24 and 48 h of culture without S9 metabolic activation, and during the last 2 h of culture with S9 metabolic activation. In the SCE assays no evidence for genotoxic activity of the fungicide was found in treatments of 24 h without and 2 h with S9. After the 24 h exposure to tolylfluanid, a weak decrease in the PI was observed. With the prolonged exposure time (48 h), dose dependence in the increase of SCE frequencies was observed. Moreover, after 48 h exposure slight fragmentation of DNA at the concentrations of 3.5 and 8.75 MUg/mL was demonstrated. SCE quantification is the most widely used approach for the assessment of genotoxic/cytogenetic effects of chemical compounds. Positive results in the assay at 48 h exposure indicated a potential of the fungicide to increase frequency of chromosomal damage (replication injuries) that is the confirmation of early effect of exposure. PMID- 21637553 TI - Assessment of the genetic risks of a metallic alloy used in medical implants. AB - The use of artificial implants provides a palliative or permanent solution for individuals who have lost some bodily function through disease, an accident or natural wear. This functional loss can be compensated for by the use of medical devices produced from special biomaterials. Titanium alloy (Ti-6Al-4V) is a well established primary metallic biomaterial for orthopedic implants, but the toxicity of the chemical components of this alloy has become an issue of concern. In this work, we used the MTT assay and micronucleus assay to examine the cytotoxicity and genotoxicity, respectively, of an extract obtained from this alloy. The MTT assay indicated that the mitochondrial activity and cell viability of CHO-K1 cells were unaffected by exposure to the extract. However, the micronucleus assay revealed DNA damage and an increase in micronucleus frequency at all of the concentrations tested. These results show that ions released from Ti-6Al-4V alloy can cause DNA and nuclear damage and reinforce the importance of assessing the safety of metallic medical devices constructed from biomaterials. PMID- 21637554 TI - Assessment of the genotoxic impact of pesticides on farming communities in the countryside of Santa Catarina State, Brazil. AB - The aim of this study was to assess the use of pesticides on farms located in the Lambedor River watershed in Guatambu, State of Santa Catarina, as well as to determine, by micronucleus testing, the risk of genotoxic impact. Samples from locally collected Cyprinus carpio, Hypostomus punctatus, Rhamdia quelen and Oreochromis niloticus gave evidence of a mean increase in micronuclei frequency from 6.21 to 13.78 in 1,000 erythrocytes, a clear indication of the genotoxic potenciality of pesticide residues in regional dams, and their significant contribution to local environmental contamination. PMID- 21637555 TI - Evaluation of genetic damage induced by glyphosate isopropylamine salt using Tradescantia bioassays. AB - Glyphosate is noted for being non-toxic in fishes, birds and mammals (including humans). Nevertheless, the degree of genotoxicity is seriously controversial. In this work, various concentrations of a glyphosate isopropylamine salt were tested using two methods of genotoxicity assaying, viz., the pink mutation assay with Tradescantia (4430) and the comet assay with nuclei from staminal cells of the same plant. Staminal nuclei were studied in two different forms, namely nuclei from exposed plants, and nuclei exposed directly. Using the pink mutation assay, isopropylamine induced a total or partial loss of color in staminal cells, a fundamental criterion utilized in this test. Consequently, its use is not recommended when studying genotoxicity with agents that produce pallid staminal cells. The comet assay system detected statistically significant (p < 0.01) genotoxic activity by isopropylamine, when compared to the negative control in both the nuclei of treated plants and directly treated nuclei, but only the treated nuclei showed a dose-dependent increase. Average migration in the nuclei of treated plants increased, when compared to that in treated nuclei. This was probably due, either to the permanence of isopropylamine in inflorescences, or to the presence of secondary metabolites. In conclusion, isopropylamine possesses strong genotoxic activity, but its detection can vary depending on the test systems used. PMID- 21637556 TI - Protective effect of carboxymethyl-glucan (CM-G) against DNA damage in patients with advanced prostate cancer. AB - Carboxymethyl-glucan (CM-G) is a soluble derivative from Saccharomyces cerevisiae (1 -> 3)(1 -> 6)-beta-D-glucan. The protective efficiency of CM-G against DNA damage in cells from patients with advanced prostate cancer (PCa), and undergoing Androgen Deprivation Therapy (ADT), was evaluated. DNA damage scores were obtained by the comet assay, both before and after treatment with CM-G. The reduction in DNA damage, ranging from 18% to 87%, with an average of 59%, was not related to the increased number of leukocytes in peripheral blood. The results demonstrate for the first time the protective effect of CM-G against DNA damage in patients with advanced PCa. Among smokers, three presented the highest reduction in DNA damage after treatment with CM-G. There was no observable relationship between DNA damage scores before and after treatment, and age, alcoholism and radiotherapy. PMID- 21637557 TI - A transcriptome analysis of mitten crab testes (Eriocheir sinensis). AB - The identification of expressed genes involved in sexual precocity of the mitten crab (Eriocheir sinensis) is critical for a better understanding of its reproductive development. To this end, we constructed a cDNA library from the rapid developmental stage of testis of E. sinensis and sequenced 3,388 randomly picked clones. After processing, 2,990 high-quality expressed sequence tags (ESTs) were clustered into 2,415 unigenes including 307 contigs and 2,108 singlets, which were then compared to the NCBI non-redundant (nr) protein and nucleotide (nt) database for annotation with Blastx and Blastn, respectively. After further analysis, 922 unigenes were obtained with concrete annotations and 30 unigenes were found to have functions possibly related to the process of reproduction in male crabs - six transcripts relevant to spermatogenesis (especially Cyclin K and RecA homolog DMC1), two transcripts involved in nuclear protein transformation, two heat-shock protein genes, eleven transcription factor genes (a series of zinc-finger proteins), and nine cytoskeleton protein-related genes. Our results, besides providing valuable information related to crustacean reproduction, can also serve as a base for future studies of reproductive and developmental biology. PMID- 21637558 TI - The history of the introduction of the giant river prawn, Macrobrachium cf. rosenbergii (Decapoda, Palaemonidae), in Brazil: New insights from molecular data. AB - The giant river prawn, Macrobrachium cf. rosenbergii, is one of the most cultivated freshwater prawns in the world and has been introduced into more than 40 countries. In some countries, this prawn is considered an invasive species that requires close monitoring. Recent changes in the taxonomy of this species (separation of M. rosenbergii and M. dacqueti) require a re-evaluation of introduced taxa. In this work, molecular analyses were used to determine which of these two species was introduced into Brazil and to establish the geographic origin of the introduced populations that have invaded Amazonian coastal waters. The species introduced into Brazil was M. dacqueti through two introduction events involving prawns originating from Vietnam and either Bangladesh or Thailand. These origins differ from historical reports of the introductions and underline the need to confirm the origin of other exotic populations around the world. The invading populations in Amazonia require monitoring not only because the biodiversity of this region may be affected by the introduction, but also because admixture of different native haplotypes can increase the genetic variability and the likelihood of persistence of the invading species in new habitats. PMID- 21637559 TI - Genetic structure of the snakehead murrel, Channa striata (channidae) based on the cytochrome c oxidase subunit I gene: Influence of historical and geomorphological factors. AB - Nucleotide sequences of a partial cytochrome c oxidase subunit I gene were used to assess the manner in which historical processes and geomorphological effects may have influenced genetic structuring and phylogeographic patterns in Channa striata. Assaying was based on individuals from twelve populations in four river systems, which were separated into two regions, the eastern and western, of the biodiversely rich state of Perak in central Peninsular Malaysia. In 238 specimens, a total of 368-bp sequences with ten polymorphic sites and eleven unique haplotypes were detected. Data on all the twelve populations revealed incomplete divergence due to past historical coalescence and the short period of separation. Nevertheless, SAMOVA and F(ST) revealed geographical structuring existed to a certain extent in both regions. For the eastern region, the data also showed that the upstream populations were genetically significantly different compared to the mid- and downstream ones. It is inferred that physical barriers and historical processes played a dominant role in structuring the genetic dispersal of the species. A further inference is that the Grik, Tanjung Rambutan and Sungkai are potential candidates for conservation and aquaculture programmes since they contained most of the total diversity in this area. PMID- 21637560 TI - Genetic evaluation of the mating system in the blue-and-yellow macaw (Ara ararauna, Aves, Psittacidae) by DNA fingerprinting. AB - More than 90% of birds are socially monogamous, although genetic studies indicate that many are often not sexually monogamous. In the present study, DNA fingerprinting was used to estimate the genetic relationships between nestlings belonging to the same broods to evaluate the mating system in the socially monogamous macaw, Ara ararauna. We found that in 10 of 11 broods investigated, the nestlings showed genetic similarity levels congruent with values expected among full-sibs, suggesting that they shared the same parents. However, in one brood, the low genetic similarity observed between nestlings could be a result of intraspecific brood parasitism, intraspecific nest competition or extra-pair paternity. These results, along with available behavioral and life-history data, imply that the blue-and-yellow macaw is not only socially, but also genetically monogamous. However, the occurrence of eventual cases of extra-pair paternity cannot be excluded. PMID- 21637561 TI - Microsatellite characterization of Cimarron Uruguayo dogs. AB - Various genetic markers, including microsatellites, have been used to analyze the genetic polymorphism and heterozygosity in canine breeds. In this work, we used nine microsatellite markers to investigate the genetic variability in Cimarron Uruguayo dogs, the only officially recognized native canine breed in Uruguay. DNA from 30 Cimarron Uruguayo dogs from northeastern and southern Uruguay was analyzed. The allelic frequencies for each microsatellite, the genetic variability and the consanguinity were calculated, as were the polymorphic information content (PIC) and the probability of exclusion (PE). All of the microsatellites studied were polymorphic. FH 2361, FH 2305 and PEZ 03 were the most informative, with PIC values > 0.7, in agreement with results for other canine breeds. The PE values for the markers were within the ranges previously described and were generally greater for microsatellites with higher PIC values. The heterozygosity value (0.649) was considered high since only nine microsatellites were analyzed. Compared with data for other breeds, the results obtained here indicate that Cimarron Uruguayo dogs have high genetic diversity. PMID- 21637562 TI - Genetic characterization of 18 novel microsatellite loci in northern pike (Esox lucius L.). AB - The northern pike (Esox lucius L.), an important predatory freshwater species, is undergoing significant population decline. In this study, 18 novel polymorphic microsatellite loci were isolated and used for assessing genetic variation in the Chinese Ulungur and Hungarian Balaton populations of the species. The number of alleles ranged from 2 to 13, observed heterozygosity from 0.154 to 0.920 and expected heterozygosity from 0.145 to 0.921, thereby indicating the specific usefulness of these suites of markers for investigating genetic variability. PMID- 21637563 TI - Informative microsatellites for genetic population studies of black-faced lion tamarins (Leontopithecus caissara). AB - Leontopithecus caissara is a critically endangered primate species from the Brazilian Atlantic Forest. Nineteen microsatellite loci, previously developed for congeneric species, were tested with 34 L. caissara individuals from Superagui Island. Of the 19 loci, 17 (89.4%) produced robust alleles, nine (47.4%) of these proved to be polymorphic, with a total of 23 alleles and an average of 2.56 alleles per locus. Expected and observed heterozygosity averaged 0.483 and 0.561, respectively. The exclusion power for identifying the first parent of an arbitrary offspring was 0.315 over all loci. The results thus indicate both the usefulness and limitations of these nine microsatellite loci in the genetic analysis of L. caissara, as well as their potentiality for genetic investigation in other congeneric species. PMID- 21637565 TI - Short-tandem repeat analysis in seven Chinese regional populations. AB - In the present study, we investigated the application of 13 short tandem repeat (STR) loci (D13S317, D7S820, TH01, D16S539, CSFIPO, VWA, D8S1179, TPOX, FGA, D3S1358, D21S11, D18S51 and D5S818) routinely used in forensic analysis, for delineating population relationships among seven human populations representing the two major geographic groups, namely the southern and northern Chinese. The resulting single topology revealed pronounced geographic and population partitioning, consistent with the differences in geographic location, languages and eating habits. These findings suggest that forensic STR loci might be particularly powerful tools in providing the necessary fine resolution for reconstructing recent human evolutionary history. PMID- 21637564 TI - Mucopolysaccharidosis I, II, and VI: Brief review and guidelines for treatment. AB - Mucopolysaccharidoses (MPS) are rare genetic diseases caused by the deficiency of one of the lysosomal enzymes involved in the glycosaminoglycan (GAG) breakdown pathway. This metabolic block leads to the accumulation of GAG in various organs and tissues of the affected patients, resulting in a multisystemic clinical picture, sometimes including cognitive impairment. Until the beginning of the XXI century, treatment was mainly supportive. Bone marrow transplantation improved the natural course of the disease in some types of MPS, but the morbidity and mortality restricted its use to selected cases. The identification of the genes involved, the new molecular biology tools and the availability of animal models made it possible to develop specific enzyme replacement therapies (ERT) for these diseases. At present, a great number of Brazilian medical centers from all regions of the country have experience with ERT for MPS I, II, and VI, acquired not only through patient treatment but also in clinical trials. Taking the three types of MPS together, over 200 patients have been treated with ERT in our country. This document summarizes the experience of the professionals involved, along with the data available in the international literature, bringing together and harmonizing the information available on the management of these severe and progressive diseases, thus disclosing new prospects for Brazilian patients affected by these conditions. PMID- 21637566 TI - The insulin polymorphism -23Hph increases the risk for type 1 diabetes mellitus in the Romanian population. AB - The insulin -23Hph and IGF2 Apa polymorphisms were genotyped in Romanian patients with T1DM (n = 204), T2DM (n = 215) or obesity (n = 200) and normoponderal healthy subjects (n = 750). The genotypes of both polymorphisms were distributed in concordance with Hardy-Weinberg equilibrium in all groups. The -23Hph AA genotype increased the risk for T1DM (OR: 3.22, 95%CI: 2.09-4.98, p < 0,0001), especially in patients without macroalbuminuria (OR: 4.32, 95%CI: 2.54-7.45, p < 0,0001). No other significant association between the alleles or genotypes of insulin -23Hph and IGF2 Apa and diabetes or obesity was identified. PMID- 21637567 TI - Polymorphisms in promoter sequences of MDM2, p53, and p16 genes in normal Japanese individuals. AB - Research has been conducted to identify sequence polymorphisms of gene promoter regions in patients and control subjects, including normal individuals, and to determine the influence of these polymorphisms on transcriptional regulation in cells that express wild-type or mutant p53. In this study we isolated genomic DNA from whole blood of healthy Japanese individuals and sequenced the promoter regions of the MDM2, p53, and p16(INK4a) genes. We identified polymorphisms comprising 3 nucleotide substitutions at exon 1 and intron 1 regions of the MDM2 gene and 1 nucleotide insertion at a poly(C) nucleotide position in the p53 gene. The Japanese individuals also exhibited p16(INK4a) polymorphisms at several positions, including position -191. Reporter gene analysis by using luciferase revealed that the polymorphisms of MDM2, p53, and p16(INK4a) differentially altered luciferase activities in several cell lines, including the Colo320DM, U251, and T98G cell lines expressing mutant p53. Our results indicate that the promoter sequences of these genes differ among normal Japanese individuals and that polymorphisms can alter gene transcription activity. PMID- 21637568 TI - Interactions of polymorphisms in different clock genes associated with circadian phenotypes in humans. AB - Several studies have shown that mutations and polymorphisms in clock genes are associated with abnormal circadian parameters in humans and also with more subtle non-pathological phenotypes like chronotypes. However, there have been conflicting results, and none of these studies analyzed the combined effects of more than one clock gene. Up to date, association studies in humans have focused on the analysis of only one clock gene per study. Since these genes encode proteins that physically interact with each other, combinations of polymorphisms in different clock genes could have a synergistic or an inhibitory effect upon circadian phenotypes. In the present study, we analyzed the combined effects of four polymorphisms in four clock genes (Per2, Per3, Clock and Bmal1) in people with extreme diurnal preferences (morning or evening). We found that a specific combination of polymorphisms in these genes is more frequent in people who have a morning preference for activity and there is a different combination in individuals with an evening preference for activity. Taken together, these results show that it is possible to detect clock gene interactions associated with human circadian phenotypes and bring an innovative idea of building a clock gene variation map that may be applied to human circadian biology. PMID- 21637569 TI - In silico analysis of alpha1-antitrypsin variants: The effects of a novel mutation. AB - Alpha1-antitrypsin (AAT) is a highly polymorphic protein with more than 120 variants that are classified as normal (normal protein secretion), deficient (reduced circulating AAT level caused by defective secretion) or null (no protein secretion). Alpha1-antitrypsin deficiency, one of the most common genetic disorders, predisposes adults to pulmonary emphysema and, to a lesser extent, chronic liver disease and cirrhosis. In this report, we provide additional sequence data for alpha1-antitrypsin based on the characterization of a novel variant detected in a 53-year-old heterozygous patient with chronic obstructive pulmonary disease. The mutation occurred on a PI*M2 base allele and was characterized by a T -> C transition at nt 97 in exon II that led to the replacement of phenylalanine by leucine (F33L). Since the mutation was found in the heterozygous state with the expression of a normally secreted variant (PI*M1) it was not possible to assess the pattern of F33L secretion. However, computational analyses based on evolutionary, structural and functional information indicated a reduction of 23 A (3) in the side chain volume and the creation of a cavity in the protein hydrophobic core that likely disturbed the tridimensional structure and folding of AAT. The accuracy of the in silico prediction was confirmed by testing known mutations. PMID- 21637570 TI - DNA repair genes XRCC1 and XRCC3 polymorphisms and their relationship with the level of micronuclei in breast cancer patients. AB - Breast cancer (BC) is the most prevalent type worldwide, besides being one of the most common causes of death among women. It has been suggested that sporadic BC is most likely caused by low-penetrance genes, including those involved in DNA repair mechanisms. Furthermore, the accumulation of DNA damage may contribute to breast carcinogenesis. In the present study, the relationship between two DNA repair genes, viz., XRCC1 (Arg399Gln) and XRCC3 (Thr241Met) polymorphisms, and the levels of chromosome damage detected in 65 untreated BC women and 85 healthy controls, was investigated. Chromosome damage was evaluated through micronucleus assaying, and genotypes determined by PCR-RFLP methodology. The results showed no alteration in the risk of BC and DNA damage brought about by either XRCC1 (Arg399Gln) or XRCC3 (Thr241Met) action in either of the two groups. Nevertheless, on evaluating BC risk in women presenting levels of chromosome damage above the mean, the XRCC3Thr241Met polymorphism was found to be more frequent in the BC group than in the control, thereby leading to the conclusion that there is a slight association between XRCC3 (241 C/T) genotypes and BC risk in the subgroups with higher levels of chromosome damage. PMID- 21637571 TI - Prevalence of common alpha-thalassemia determinants in south Brazil: Importance for the diagnosis of microcytic anemia. AB - Alpha thalassemia has not been systematically investigated in Brazil. In this study, 493 unrelated individuals from the southernmost Brazilian state of Rio Grande do Sul were screened for deletional forms of alpha-thalassemia. One hundred and one individuals had microcytic anemia (MCV < 80 fL) and a normal hemoglobin pattern (Hb A (2) < 3.5% and Hb F < 1%). The subjects were screened for - alpha(3.7) , - alpha(4.2) , - alpha(20.5) , - (SEA) and - (MED) deletions but only the - alpha(3.7) allele was detected. The - alpha(3.7) allele frequency in Brazilians of European and African ancestry was 0.02 and 0.12, respectively, whereas in individuals with microcytosis the frequency was 0.20. The prevalence of alpha-thalassemia was significantly higher in individuals with microcytosis than in healthy individuals (p = 0.001), regardless of their ethnic origin. There were also significant differences in the hematological parameters of individuals with - alpha(3.7) / alphaalpha, - alpha(3.7) /- alpha(3.7) and beta-thalassemia trait compared to healthy subjects. These data suggest that alpha-thalassemia is an important cause of microcytosis and mild anemia in Brazilians. PMID- 21637572 TI - Relationship of an hRAD54 gene polymorphism (2290 C/T) in an Ecuadorian population with chronic myelogenous leukemia. AB - The hRAD54 gene is a key member of the RAD52 epistasis group involved in repair of double-strand breaks (DSB) by homologous recombination (HR). Thus, alterations of the normal function of these genes could generate genetic instability, shifting the normal process of the cell cycle, leading the cells to develop into cancer. In this work we analyzed exon 18 of the hRAD54 gene, which has been previously reported by our group to carry a silent polymorphism, 2290 C/T (Ala730Ala), associated to meningiomas. We performed a PCR-SSCP method to detect the polymorphism in 239 samples including leukemia and normal control population. The results revealed that the 2290 C/T polymorphism has frequencies of 0.1 for the leukemia and 0.1 for the control group. These frequencies show no statistical differences. Additionally, we dissected the leukemia group in chronic myelogenous leukemia (CML) and acute lymphoblastic leukemia (ALL) to evaluate the polymorphism. The frequencies found in these subgroups were 0.14 for CML and 0.05 for ALL. We found statistically significant differences between CML patients and the control group (p < 0.05) but we did not find significant differences between ALL and the control group (p > 0.05). These results suggest a possible link between the 2290 C/T polymorphism of the hRAD54 gene and CML. PMID- 21637573 TI - Molecular characterization and genetic structure of the Nero Siciliano pig breed. AB - Nero Siciliano is an autochthonous pig breed that is reared mainly in semi extensive systems in northeastern Sicily. Despite its economic importance and well-appreciated meat products, this breed is currently endangered. Consequently, an analysis of intra-breed variability is a fundamental step in preserving this genetic resource and its breeding system. In this work, we used 25 microsatellite markers to examine the genetic composition of 147 unrelated Nero Siciliano pigs. The total number of alleles detected (249, 9.96 per locus) and the expected heterozygosity (0.708) indicated that this breed had a high level of genetic variability. Bayesian cluster analysis showed that the most likely number of groups into which the sample could be partitioned was nine. Based on the proportion of each individuals genome derived from ancestry, pigs with at least 70% of their genome belonging to one cluster were assigned to that cluster. The cluster size ranged from 7 to 17 (n = 108). Genetic variability in this sub population was slightly lower than in the whole sample, genetic differentiation among clusters was moderate (F(ST) 0.125) and the F(IS) value was 0.011. NeighborNet and correspondence analysis revealed two clusters as the most divergent. Molecular coancestry analysis confirmed the good within-breed variability and highlighted the clusters that retained the highest genetic diversity. PMID- 21637574 TI - Change in genetic size of small-closed populations: Lessons from a domestic mammal population. AB - The aim of this study was to monitor changes in genetic size of a small-closed population of Iranian Zandi sheep, by using pedigree information from animals born between 1991 and 2005. The genetic size was assessed by using measures based on the probability of identity-by-descend of genes (coancestry, f, and effective population size, N(e) ), as well as measures based on probability of gene origin (effective number of founders, f(e) , effective number of founder genomes, f(g) , and effective number of non-founder genomes, f(ne) ). Average coancestry, or the degree of genetic similarity of individuals, increased from 0.81% to 1.44% during the period 1993 to 2005, at the same time that N(e) decreased from 263 to 93. The observed trend for f(e) was irregular throughout the experiment in a way that f(e) was 68, 87, 77, 92, and 80 in 1993, 1996, 1999, 2002, and 2005, respectively. Simultaneously, f(g) , the most informative effective number, decreased from 61 to 35. The index of genetic diversity (GD) which was obtained from estimates of f(g) , decreased about 2% throughout the period studied. In addition, a noticeable reduction was observed in the estimates of f(ne) from 595 in 1993 to 61 in 2005. The higher than 1 ratio of f(e) to f(g) indicated the presence of bottlenecks and genetic drift in the development of this population of Zandi sheep. From 1993 to 1999, f(ne) was much higher than f(e) , thereby indicating that with respect to loss of genetic diversity, the unequal contribution of founders was more important than the random genetic drift in non founder generations. Subsequently, random genetic drift in non-founder generations was the major reason for f(e) > f(ne) . The minimization of average coancestry in new reproductive individuals was recommended as a means of preserving the population against a further loss in genetic diversity. PMID- 21637575 TI - Chromosomes of Theridiidae spiders (Entelegynae): Interspecific karyotype diversity in Argyrodes and diploid number intraspecific variability in Nesticodes rufipes. AB - Theridiidae is a derived family within the Araneoidea clade. In contrast to closely related groups, the 2n(male) = 20+X(1) X (2) with acro/telocentric chromosomes is the most widespread karyotype among the theridiid spiders. In this work, the cytogenetic analysis of Argyrodes elevatus revealed original chromosome features different from those previously registered for Theridiidae, including the presence of 2n(male) = 20+X with meta/submetacentric chromosomes. Most individuals of Nesticodes rufipes showed family conserved karyotype characteristics. However, one individual had a 2n(male) = 24 due to the presence of an extra chromosome pair, which exhibited regular behavior and reductional segregation during meiosis. After silver staining, mitotic cells exhibited NORs localized on the terminal regions of the short arms of pairs 2, 3, and 4 of A. elevatus and on the terminal regions of long arms of pair 4 of N. rufipes. The comparative analysis with data from phylogenetically related species allowed the clarification of the origin of the interspecific and intraspecific chromosome variability observed in Argyrodes and in N. rufipes, respectively. PMID- 21637576 TI - Sociogenetic structure of Polistes (Aphanilopterus) versicolor Olivier, 1791 colonies (Hymenoptera, Vespidae, Polistini). AB - The observation of two distinct, well-defined oviposition areas in nests of the primitively eusocial wasp Polistes versicolor suggests the presence of multiple egg-layers and territorial behaviors. Electrophoretic analysis of enzyme loci in pupae from 35 colonies revealed an average observed heterozygosity of 0.10 and the existence of private polymorphisms, thereby indicating a low dispersion in this species. No evidence of diploid males was found. Phenotypic segregation analysis revealed the presence of more than one egg-laying female in 15 out of 35 colonies, as well as spatially preferential oviposition in 2 out of 13 nests, with distinct oviposition areas. Genetic relatedness estimates for brood were lower than expected for haplodiploid species under monogynous conditions (r = 0.75 for female broods and r = 0.5 for male) in 4 of those 13 nests, thereby inferring complex sociogenetic structuring in Polistes versicolor colonies. PMID- 21637577 TI - Isolation and characterization of genes functionally involved in ovarian development of the giant tiger shrimp Penaeus monodon by suppression subtractive hybridization (SSH). AB - Suppression subtractive hybridization (SSH) libraries between cDNA in stages I (previtellogenic) and III (cortical rod) ovaries of the giant tiger shrimp (Penaeus monodon) were established. In all, 452 ESTs were unidirectionally sequenced. Sequence assembly generated 28 contigs and 201 singletons, 109 of which (48.0%) corresponding to known sequences previously deposited in GenBank. Several reproduction-related transcripts were identified. The full-length cDNA of anaphase promoting complex subunit 11 (PmAPC11; 600 bp with an ORF of 255 bp corresponding to a polypeptide of 84 amino acids) and selenoprotein Mprecursor (PmSePM; 904 bp with an ORF of 396 bp corresponding to a polypeptide of 131 amino acids) were characterized and reported for the first time in penaeid shrimp. Semiquantitative RT-PCR revealed that the expression levels of PmSePM and keratinocyte-associated protein 2 significantly diminished throughout ovarian development, whereas Ser/Thrcheckpoint kinase 1 (Chk1), DNA replication licensing factor mcm2 and egalitarian were down-regulated in mature ovaries of wild P. monodon (p < 0.05). Accordingly, the expression profiles of PmSePM and keratinocyte-associated protein 2 could be used as biomarkers for evaluating the degree of reproductive maturation in domesticated P. monodon. PMID- 21637578 TI - Cloning and endogenous expression of a Eucalyptus grandis UDP-glucose dehydrogenase cDNA. AB - UDP-glucose dehydrogenase (UGDH) catalyzes the oxidation of UDP-glucose (UDP-Glc) to UDP-glucuronate (UDP-GlcA), a key sugar nucleotide involved in the biosynthesis of plant cell wall polysaccharides. A full-length cDNA fragment coding for UGDH was cloned from the cambial region of 6-month-old E. grandis saplings by RT-PCR. The 1443-bp-ORF encodes a protein of 480 amino acids with a predicted molecular weight of 53 kDa. The recombinant protein expressed in Escherichia coli catalyzed the conversion of UDP-Glc to UDP-GlcA, confirming that the cloned cDNA encodes UGDH. The deduced amino acid sequence of the cDNA showed a high degree of identity with UGDH from several plant species. The Southern blot assay indicated that more than one copy of UGDH is present in Eucalyptus. These results were also confirmed by the proteomic analysis of the cambial region of 3- and 22-year-old E. grandis trees by 2-DE and LC-MS/MS, showing that at least two isoforms are present. The cloned gene is mainly expressed in roots, stem and bark of 6-month-old saplings, with a lower expression in leaves. High expression levels were also observed in the cambial region of 3- and 22-year-old trees. The results described in this paper provide a further view of the hemicellulose biosynthesis during wood formation in E. grandis. PMID- 21637579 TI - Population structures of Brazilian tall coconut (Cocos nucifera L.) by microsatellite markers. AB - Coconut palms of the Tall group were introduced to Brazil from the Cape Verde Islands in 1553. The present study sought to evaluate the genetic diversity among and within Brazilian Tall coconut populations. Samples were collected of 195 trees from 10 populations. Genetic diversity was accessed by investigating 13 simple sequence repeats (SSR) loci. This provided a total of 68 alleles, ranging from 2 to 13 alleles per locus, with an average of 5.23. The mean values of gene diversity (H(e) ) and observed heterozygosity (H(o) ) were 0.459 and 0.443, respectively. The genetic differentiation among populations was estimated at theta^P=0.1600and the estimated apparent outcrossing rate was t(a) = 0.92. Estimates of genetic distances between the populations varied from 0.034 to 0.390. Genetic distance and the corresponding clustering analysis indicate the formation of two groups. The first consists of the Baia Formosa, Georgino Avelino, and Sao Jose do Mipibu populations and the second consists of the Japoata, Pacatuba, and Praia do Forte populations. The correlation matrix between genetic and geographic distances was positive and significant at a 1% probability. Taken together, our results suggest a spatial structuring of the genetic variability among the populations. Geographically closer populations exhibited greater similarities. PMID- 21637580 TI - In silicio expression analysis of PKS genes isolated from Cannabis sativa L. AB - Cannabinoids, flavonoids, and stilbenoids have been identified in the annual dioecious plant Cannabis sativa L. Of these, the cannabinoids are the best known group of this plant's natural products. Polyketide synthases (PKSs) are responsible for the biosynthesis of diverse secondary metabolites, including flavonoids and stilbenoids. Biosynthetically, the cannabinoids are polyketide substituted with terpenoid moiety. Using an RT-PCR homology search, PKS cDNAs were isolated from cannabis plants. The deduced amino acid sequences showed 51% 73% identity to other CHS/STS type sequences of the PKS family. Further, phylogenetic analysis revealed that these PKS cDNAs grouped with other non chalcone-producing PKSs. Homology modeling analysis of these cannabis PKSs predicts a 3D overall fold, similar to alfalfa CHS2, with small steric differences on the residues that shape the active site of the cannabis PKSs. PMID- 21637581 TI - Genome re-assignment of Arachis trinitensis (Sect. Arachis, Leguminosae) and its implications for the genetic origin of cultivated peanut. AB - The karyotype structure of Arachis trinitensis was studied by conventional Feulgen staining, CMA/DAPI banding and rDNA loci detection by fluorescence in situ hybridization (FISH) in order to establish its genome status and test the hypothesis that this species is a genome donor of cultivated peanut. Conventional staining revealed that the karyotype lacked the small "A chromosomes" characteristic of the A genome. In agreement with this, chromosomal banding showed that none of the chromosomes had the large centromeric bands expected for A chromosomes. FISH revealed one pair each of 5S and 45S rDNA loci, located in different medium-sized metacentric chromosomes. Collectively, these results suggest that A. trinitensis should be removed from the A genome and be considered as a B or non-A genome species. The pattern of heterochromatic bands and rDNA loci of A. trinitensis differ markedly from any of the complements of A. hypogaea, suggesting that the former species is unlikely to be one of the wild diploid progenitors of the latter. PMID- 21637582 TI - Exploiting a wheat EST database to assess genetic diversity. AB - Expressed sequence tag (EST) markers have been used to assess variety and genetic diversity in wheat (Triticum aestivum). In this study, 1549 ESTs from wheat infested with yellow rust were used to examine the genetic diversity of six susceptible and resistant wheat cultivars. The aim of using these cultivars was to improve the competitiveness of public wheat breeding programs through the intensive use of modern, particularly marker-assisted, selection technologies. The F(2) individuals derived from cultivar crosses were screened for resistance to yellow rust at the seedling stage in greenhouses and adult stage in the field to identify DNA markers genetically linked to resistance. Five hundred and sixty ESTs were assembled into 136 contigs and 989 singletons. BlastX search results showed that 39 (29%) contigs and 96 (10%) singletons were homologous to wheat genes. The database-matched contigs and singletons were assigned to eight functional groups related to protein synthesis, photosynthesis, metabolism and energy, stress proteins, transporter proteins, protein breakdown and recycling, cell growth and division and reactive oxygen scavengers. PCR analyses with primers based on the contigs and singletons showed that the most polymorphic functional categories were photosynthesis (contigs) and metabolism and energy (singletons). EST analysis revealed considerable genetic variability among the Turkish wheat cultivars resistant and susceptible to yellow rust disease and allowed calculation of the mean genetic distance between cultivars, with the greatest similarity (0.725) being between Harmankaya99 and Sonmez2001, and the lowest (0.622) between Aytin98 and Izgi01. PMID- 21637583 TI - Karyotype differentiation in three species of Tripogandra Raf. (Commelinaceae) with different ploidy levels. AB - Most species of the genus Tripogandra (Commelinaceae) are taxonomically poorly circumscribed, in spite of having a relatively stable basic number x = 8. Aiming to estimate the cytological variation among Tripogandra species carrying this base number, several structural karyotypic characters were investigated in the diploid T. glandulosa, the hexaploid T. serrulata, and the octoploid T. diuretica. A careful evaluation of chromosome size and morphology did not reveal clear chromosome homeologies among karyotypes. The mean chromosome size was strongly reduced in the octoploid species, but not in the hexaploid species. They also differed largely in the CMA(+) banding pattern and in the number of 5S and 45S rDNA sites per monoploid chromosome complement. All three species showed proximal DAPI (+) heterochromatin, although in T. serrulata this kind of heterochromatin was only visible after FISH. Further, the meiosis in T. serrulata was highly irregular, suggesting that this species has a hybrid origin. The data indicate that, in spite of the conservation of the base number, these species are karyologically quite different from each other. PMID- 21637584 TI - Molecular cloning and expression of key gene encoding hypothetical DNA polymerase from B. mori parvo-like virus. AB - BmPLV-Z is the abbreviation for Bombyx mori parvo-like virus (China isolate). This is a novel virus with two single-stranded linear DNA molecules, viz., VD1 (6543 bp) and VD2 (6022 bp), which are encapsidated respectively into separate virions. Analysis of the deduced amino acid sequence of VD1-ORF4 indicated the existence of a putative DNA-polymerase with exonuclease activity, possibly involved in the replication of BmPLV-Z. In the present study, a recombinant baculovirus was constructed to express the full length of the protein encoded by the VD1-ORF4 gene (3318 bp). In addition, a 2163-bp fragment amplified from the very same gene was cloned into prokaryotic expression vector pET-30a and expressed in E.coli Rosetta 2 (DE3) pLysS. The expressed fusion protein was employed to immunize New Zealand white rabbits for the production of an antiserum, afterwards used for examining the expression of the protein encoded by VD1-ORF4 gene in Sf-9 cells infected with recombinant baculovirus. Western blot analysis of extracts from thus cells infected revealed a specific band of about 120 kDa, thereby indicating that the full length protein encoded by the VD1-ORF4 gene had been successfully and stably expressed in Sf-9 cells. PMID- 21637585 TI - Phylogenetic position of an uncharacterized Brazilian strain of bovine papillomavirus in the genus Xipapillomavirus based on sequencing of the L1 open reading frame. AB - The use of PCR assays with degenerate primers has suggested the existence of numerous as yet uncharacterized bovine papillomaviruses (BPV). Despite the endemic nature of BPV infections, the identification of BPV types in Brazilian cattle is still only sporadic. However, in a recent analysis of a partial segment of the L1 gene, we observed notable diversity among the BPV types detected. The aim of this study was to determine the phylogenetic position of the previously identified wild strain BPV/BR-UEL2 detected in the state of Parana in Brazil. Since previous analysis of the partial L1 sequence had shown that this strain was most closely related to BPV type 4, genus-specific primers were designed. Phylogenetic analysis using complete L1 ORF sequences revealed that BPV/BR-UEL2 was related to BPV types classified in the genus Xipapillomavirus and shared the highest L1 nucleotide sequence similarity with BPV type 4 (78%). This finding suggests that BPV/BR-UEL2 should be classified as a potential new type of BPV in the genus Xipapillomavirus. PMID- 21637586 TI - Toxicity and genotoxicity in Astyanax bimaculatus (Characidae) induced by microcystins from a bloom of Microcystis spp. AB - Studies of genotoxicity in fish caused by cyanobacterial microcystins can be useful both in determining the sensitivity of native species, as well as comparing exposure routes. The genotoxicity caused by the microcystins LR and LA from a bloom collected in a eutrophic lake, was revealed in the fish Astyanaxbimaculatus, a native species from South America. LC50 (72 h) was determined as 242.81 MUg L (-1) and LD50 (72 h) as 49.19 MUg kg (-1) bw. There was a significant increase of DNA damage in peripheral erythrocytes, following intraperitoneal injection (ip) with tested concentrations of 24.58 MUg kg (-1) bw and 36.88 MUg kg (-1) bw, as well as through body exposure to a concentration of 103.72 MUg L (-1) . Micronucleus (MN) induction was observed after ip injections of 24.58 MUg kg (-1) bw and 36.88 MUg kg (-1) bw for 72 h, as well as following body exposure for 72 at 103.72 MUg L (-1) . Thus, both exposure routes resulted in MN induction and DNA damage. Apoptosis-necrosis testing was carried out only by ip injection with concentrations of 24.58 MUg kg (-1) bw and 36.88 MUg kg- 1 bw. Exposure to microcystins at lower concentrations induced more apoptosis than necrosis in peripheral erythrocytes, whereas exposure at higher concentrations gave rise to both conditions. Thus, Astyanax bimaculatus can be considered as a species sensitive to the genotoxic effects caused by microcystins. PMID- 21637587 TI - Micronuclei induced by reverse transcriptase inhibitors in mononucleated and binucleated cells as assessed by the cytokinesis-block micronucleus assay. AB - This study evaluated the clastogenic and/or aneugenic potential of three nucleoside reverse transcriptase inhibitors (zidovudine - AZT, lamivudine - 3TC and stavudine - d4T) using the cytokinesis-block micronucleus (CBMN) assay in human lymphocyte cultures. All three inhibitors produced a positive response when tested in binucleated cells. The genotoxicity of AZT and 3TC was restricted to binucleated cells since there was no significant increase in the frequency of micronuclei in mononucleated cells. This finding indicated that AZT and 3TC caused chromosomal breakage and that their genotoxicity was related to a clastogenic action. In addition to the positive response observed with d4T in binucleated cells, this drug also increased the frequency of micronuclei in mononucleated cells, indicating clastogenic and aneugenic actions. Since the structural differences between AZT and 3TC and AZT and d4T involve the 3' position in the 2'-deoxyribonucleoside and in an unsaturated 2',3',dideoxyribose, respectively, we suggest that an unsaturated 2', 3', dideoxyribose is responsible for the clastogenic and aneugenic actions of d4T. PMID- 21637588 TI - A comparison of the effects of physical and chemical mutagens in sesame (Sesamum indicum L.). AB - Three sesame genotypes (Rama, SI 1666 and IC 21706) were treated with physical (gamma-rays: 200 Gy, 400 Gy or 600 Gy) or chemical (ethyl methane sulphonate, EMS: 0.5%, 1.0%, 1.5% or 2.0%) mutagens and their mutagenic effectiveness and efficiency were estimated in the M (2) generation. The M (3) generation was used to identify the most effective mutagen and dose for induction of mutations. The average effectiveness of EMS was much higher than gamma-rays. The lowest dose of gamma-rays (200 Gy) and the lowest concentration of EMS (0.5%) showed the highest mutagenic efficiency in all genotypes. Analysis of the M (3) generation data based on parameters such as the variance ratio and the difference in residual variances derived from the model of Montalvan and Ando indicated that 0.5% concentration of EMS was the most effective treatment for inducing mutations. PMID- 21637589 TI - Scenario of the spread of the invasive species Zaprionus indianus Gupta, 1970 (Diptera, Drosophilidae) in Brazil. AB - Zaprionus indianus was first recorded in Brazil in 1999 and rapidly spread throughout the country. We have obtained data on esterase loci polymorphisms (Est2 and Est3), and analyzed them, using Landscape Shape Interpolation and the Monmonier Maximum Difference Algorithm to discover how regional invasion occurred. Hence, it was apparent that Z. indianus, after first arriving in Sao Paulo state, spread throughout the country, probably together with the transportation of commercial fruits by way of the two main Brazilian freeways, BR 153, to the south and the surrounding countryside, and the BR 116 along the coast and throughout the north-east. PMID- 21637590 TI - Genetic structure of red-handed howler monkey populations in the fragmented landscape of Eastern Brazilian Amazonia. AB - We genotyped 15 microsatellite loci in order to evaluate the effects of habitat fragmentation, caused by flooding of the Tucurui reservoir, on the genetic structure of Alouatta belzebul in eastern Amazonia. The analysis included two populations sampled in 1984, representing both margins of the Tocantins river, and three populations sampled 18 years later. Minimal differences in the diversity levels between present-day (Ho = 0.62-0.69 and A(R) = 6.07-7.21) and pre-flooding (Ho = 0.60-0.62 and A (R) = 6.27-6.77) populations indicated there was no significant loss of genetic variability, possibly because of successful management strategies applied during the flooding. The changes observed were limited to shifts in the composition of alleles, which presumably reflect the admixture of subpopulations during flooding. Given this, there were significant differences in the Rst values (p = 0.05) in all but one between-site comparison. Both present-day and original populations showed a deficit of heterozygotes, which suggests that this may be typical of the species, at least at a local level, perhaps because of specific ecological characteristics. The relatively large number of private alleles recorded in all populations may be a consequence of the Wahlund effect resulting from population admixture or a process of expansion rather than the loss of rare alleles through genetic drift. Additionally, the levels of genetic variability observed in this study were higher than those reported for other species of Neotropical primates, suggesting good fitness levels in these A. belzebul populations. Regular genetic monitoring of remnant populations, especially on islands, should nevertheless be an integral component of long-term management strategies. PMID- 21637591 TI - Genetic variability in five populations of Partamona helleri (Hymenoptera, Apidae) from Minas Gerais State, Brazil. AB - Partamona is a Neotropical genus of stingless bees that comprises 33 species distributed from Mexico to southern Brazil. These bees are well-adapted to anthropic environments and build their nests in several substrates. In this study, 66 colonies of Partamona helleri from five localities in the Brazilian state of Minas Gerais (Sao Miguel do Anta, Teixeiras, Porto Firme, Vicosa and Rio Vermelho) were analyzed using nine microsatellite loci in order to assess their genetic variability. Low levels of observed (H(o) = 0.099-0.137) and expected (H (e) = 0.128-0.145) heterozygosity were encountered and revealed discrete genetic differentiation among the populations (F (ST) = 0.025). AMOVA further showed that most of the total genetic variation (94.24%) in P. helleri was explained by the variability within local populations. PMID- 21637592 TI - Testing the Rio Doce as a riverine barrier in shaping the Atlantic rainforest population divergence in the rodent Akodon cursor. AB - Akodon cursor occurs in dense rainforest from northern (8 degrees S) to southern (26 degrees S) states along the Atlantic coast of Brazil. Previous karyological and molecular data revealed two major clades, one including northern (8-15 degrees S) and the other southern (19-26 degrees S) populations. The center of geographic distribution (15-20 degrees S), which included the state of Espirito Santo, was identified as a potential vicariance region. Since river barriers are among the most discussed models in the study of Neotropical diversification, we examined whether the Rio Doce (19 degrees S) plays an important role in shaping the population genetic divergence of A. cursor by including samples from Espirito Santo in the analysis. Our results showed that the northern-southern division region in Atlantic forest was no coincidence with the presence of the Rio Doce by refuting the hypothesis that this river is an effective barrier to gene flow between populations. Instead, we found evidence that isolation by geographical distance shaped the phylogeographical structure in the southern lineage. However, there is uncertainty about effectiveness of the processes involved and further studies based on wider sampling are needed. PMID- 21637593 TI - Expression of DLK1 and MEG3 genes in porcine tissues during postnatal development. AB - The Drosophila-like homolog 1 (DLK1), a transmembrane signal protein similar to other members of the Notch/Delta/Serrate family, regulates the differentiation process in many types of mammalian cells. Callipyge sheep and DLK1 knockout mice are excellent examples of a fundamental role of the gene encoding DLK1 in muscle growth and fat deposition. DLK1 is located within co-regulated imprinted clusters (the DLK1/DIO3 domain), along with other imprinted genes. Some of these, e.g. the RNA coding MEG3 gene, presumedly interfere with DLK1 transcription. The aim of our study was to analyze DLK1 and MEG3 gene expression in porcine tissues (muscle, liver, kidney, heart, brain stem) during postnatal development. The highest expression of both DLK1 and MEG3 variant 1 (MEG3 var.1) was observed in the brain-stem and muscles, whereas that of MEG3 variant 2 (MEG3var.2) was the most abundant in muscles and the heart. During development (between 60 and 210 days of age) expression of analyzed genes was down-regulated in all the tissues. An exception was the brain- stem, where there was no significant change in MEG3 (both variants) mRNA level, and relatively little decline (2-fold) in that of DLK1 transcription. This may indicate a distinct function of the DLK1 gene in the brain-stem, when compared with other tissues. PMID- 21637594 TI - In silico identification of coffee genome expressed sequences potentially associated with resistance to diseases. AB - Sequences potentially associated with coffee resistance to diseases were identified by in silico analyses using the database of the Brazilian Coffee Genome Project (BCGP). Keywords corresponding to plant resistance mechanisms to pathogens identified in the literature were used as baits for data mining. Expressed sequence tags (ESTs) related to each of these keywords were identified with tools available in the BCGP bioinformatics platform. A total of 11,300 ESTs were mined. These ESTs were clustered and formed 979 EST-contigs with similarities to chitinases, kinases, cytochrome P450 and nucleotide binding site leucine rich repeat (NBS-LRR) proteins, as well as with proteins related to disease resistance, pathogenesis, hypersensitivity response (HR) and plant defense responses to diseases. The 140 EST-contigs identified through the keyword NBS-LRR were classified according to function. This classification allowed association of the predicted products of EST-contigs with biological processes, including host defense and apoptosis, and with molecular functions such as nucleotide binding and signal transducer activity. Fisher's exact test was used to examine the significance of differences in contig expression between libraries representing the responses to biotic stress challenges and other libraries from the BCGP. This analysis revealed seven contigs highly similar to catalase, chitinase, protein with a BURP domain and unknown proteins. The involvement of these coffee proteins in plant responses to disease is discussed. PMID- 21637595 TI - Mitochondrial DNA common deletion is not associated with thyroid, breast and colorectal tumors in Turkish patients. AB - Recently, efforts have been focused on mitochondrial DNA changes and their relation to human cancers. Among them, a 4977 bp deletion of mitochondrial DNA, named "common deletion", has been investigated in several types of tumors, with inconsistent results. In this study, we investigated the presence of the common deletion in tissues from 25 breast, 25 colorectal and 50 thyroid tumors and in the adjacent healthy tissues from Turkish patients. Samples from healthy volunteers were also evaluated for comparison. Two PCR-based methods were used for the detection of the common deletion. First, two pairs of primers were used to amplify wild-type and deleted mtDNA. Then, a highly sensitive nested-PCR was performed, to determine low amounts of deleted genomes. By the first method, wild type mtDNAs were observed in all samples, but a deletion was observed in only six thyroid samples, by using the nested-PCR method. In conclusion, the mitochondrial common deletion was very rare in our study group and did not appear to be not related with cancer. PMID- 21637596 TI - Identification of a rare p.G320R alpha-1-antitrypsin variant in emphysema and lung cancer patients. AB - The alpha-1-antitrypsin (A1AT) gene is highly polymorphic, with more than 100 genetic variants identified of which some can affect A1AT protein concentration and/or function and lead to pulmonary and/or liver disease. This study reports on the characterization of a p.G320R variant found in two patients, one with emphysema and the other with lung cancer. This variant results from a single base pair substitution in exon 4 of the A1AT gene, and has been characterized as P by isoelectric focusing. Functional evaluation of the A1AT p.G320R variant was through comparing specific trypsin inhibitory activity in two patients with pulmonary disorders, carriers of the p.G320R variant, and 19 healthy individuals, carriers of normal A1AT M variants. Results showed that specific trypsin inhibitory activity was lower in both emphysema (2.45 mU/g) and lung cancer (2.07 mU/g) patients than in carriers of the normal variants (range 2.51-3.71 mU/g). This rare A1AT variant is associated with reduced functional activity of A1AT protein. Considering that it was found in patients with severe pulmonary disorders, this variant could be of clinical significance. PMID- 21637597 TI - Analysis of the SLC4A1 gene in three Mexican patients with hereditary spherocytosis: Report of a novel mutation. AB - We analyzed the SLC4A1 gene in three Mexican patients with Hereditary Spherocytosis (HS). The promoter and all 20 exons were investigated through heteroduplex analysis and DNA sequencing. No DNA changes were detected in one of the three patients. Two well-known polymorphisms, Memphis I and the Diego-a blood group, were detected in another one. In the third, the HS phenotype could be explained by the novel 1885_1888dupCCGG mutation found in heterozygosis. This frameshift mutation is predicted to result in a truncated and unstable protein lacking normal functions. PMID- 21637598 TI - Immediate response of myocardium to pressure overload includes transient regulation of genes associated with mitochondrial bioenergetics and calcium availability. AB - Ventricular hypertrophy is one of the major myocardial responses to pressure overload (PO). Most studies on early myocardial response focus on the days or even weeks after induction of hypertrophic stimuli. Since mechanotransduction pathways are immediately activated in hearts undergoing increased work load, it is reasonable to infer that the myocardial gene program may be regulated in the first few hours. In the present study, we monitored the expression of some genes previously described in the context of myocardial hypertrophic growth by using the Northern blot technique, to estimate the mRNA content of selected genes in rat myocardium for the periods 1, 3, 6, 12 and 48 h after PO stimuli. Results revealed an immediate switch in the expression of genes encoding alpha and beta isoforms of myosin heavy chain, and up-regulation of the cardiac isoform of alpha actin. We also detected transitory gene regulation as the increase in mitochondrial cytochrome c oxidase 1 gene expression, parallel to down-regulation of genes encoding sarco(endo)plasmic reticulum Ca(+2) ATPase and sodium-calcium exchanger. Taken together, these results indicate that initial myocardial responses to increased work load include alterations in the contractile properties of sarcomeres and transitory adjustment of mitochondrial bioenergetics and calcium availability. PMID- 21637599 TI - Knockdown of E2f1 by RNA interference impairs proliferation of rat cells in vitro. AB - E2F1 plays a key role in cell-cycle regulation in mammals, since its transcription factor activity controls genes required for DNA synthesis and apoptosis. E2F1 deregulation is a common feature among different tumor types and can be a major cause of cell proliferation. Thus, blocking E2F1 expression by RNA interference represents a promising therapeutic approach. In this study, the introduction of specific short hairpin RNAs (shRNAs) reduced E2f1 expression by up to 77%, and impaired rat glioma cell proliferation by approximately 70%, as compared to control cells. Furthermore, we investigated the expression of E2f1 target genes, Cyclin A and Cyclin E. Cyclin A was found to be down-regulated, whereas Cyclin E had similar expression to control cells, indicating that gene(s) other than E2f1 control its transcription. Other E2f family members, E2f2 and E2f3, which have been classified in the same subgroup of transcriptional activators, were also analyzed. Expression of both E2f2 and E2f3 was similar to control cells, showing no cross-inactivation or up-regulation to compensate for the absence of E2f1. Nevertheless, their expression was insufficient to maintain the initial proliferation potential. Taken together, our results suggest that shE2f1 is a promising therapy to control tumor cell proliferation. PMID- 21637600 TI - Alpha-1 antitrypsin gene polymorphism in Chronic Obstructive Pulmonary Disease (COPD). AB - Alpha-1-antitrypsin (AAT) plays an important role in the pathogenesis of emphysema, the pathological lesion underlying the majority of the manifestations of Chronic Obstructive Pulmonary Disease (COPD). In this study we tested the hypothesis that common AAT polymorphisms influence the risk of developing COPDs. We investigated PiM1 (Ala213Val), PiM2 (Arg101His), PiM3 (Glu376Asp), PiS (Glu264Val) and PiZ (Glu342Lys) SERPINA1 alleles in 100 COPD patients and 200 healthy controls. No significant differences were observed in allele frequencies between COPD patients and controls, neither did haplotype analysis show significant differences between the two groups. A cross-sectional study revealed no significant relationship between common SERPINA1 polymorphisms (PiM1, PiM2, PiM3) and the emphysematous type of COPD. In addition, FEV(1) annual decline, determined during a two-year follow up period, revealed no difference among carriers of the tested polymorphisms. PMID- 21637601 TI - Identification of 2chromosome region translocated onto the W chromosome by RFLP with EST-cDNA clones in the Gensei-kouken strains of the mulberry silkworm, Bombyx mori L. AB - In silkworms, sex-limited strains are either obtained spontaneously or induced by X-rays or gamma rays. When a fragment of an autosome carrying a dominant allele of those genes responsible for certain characters is translocated onto a W chromosome, the female of the successive generations will express these phenotypic characters and sex discrimination can be facilitated. Gensei-kouken strains are sex-limited strains of silkworms developed by irradiating the pupae with gamma rays, by which a portion of the second chromosome is translocated onto the W chromosome. In these improved strains, the females are yellow-blooded and spin yellow cocoons. By using the EST-cDNA clones mapped on the Z chromosome, we identified the sex according to the polymorphic banding pattern or intensity of the signals. Furthermore, by using the clones on the second chromosome, the region of the second chromosome translocated onto the W chromosome was also defined. In both the A95 and A 96 strains selected for the present study, only the mid-portion of the second chromosome was translocated. The differences in length of the fragments translocated in these strains are discussed. PMID- 21637602 TI - Chromosomal localization and molecular marker development of the lipopolysaccharide and beta-1,3-glucan binding protein gene in the Zhikong scallop Chlamys farreri (Jones et Preston) (Pectinoida, Pectinidae). AB - Zhikong scallop Chlamys farreri (Jones et Preston) is an economically important species in China. Understanding its immune system would be of great help in controlling diseases. In the present study, an important immunity-related gene, the Lipopolysaccharide and Beta-1,3-glucan Binding Protein (LGBP) gene, was located on C. farreri chromosomes by mapping several lgbp-containing BAC clones through fluorescence in situ hybridization (FISH). Through the localization of various BAC clones, it was shown that only one locus of this gene existed in the genome of C. farreri, and that this was located on the long arm of a pair of homologous chromosomes. Molecular markers, consisting of eight single nucleotide polymorphism (SNPs) markers and one insertion-deletion (indel), were developed from the LGBP gene. Indel marker testing in an F1 family revealed slightly distorted segregation (p = 0.0472). These markers can be used to map the LGBP gene to the linkage map and assign the linkage group to the corresponding chromosome. Segregation distortion of the indel marker indicated genes with deleterious alleles might exist in the surrounding region of the LGBP gene. PMID- 21637603 TI - Differentially expressed genes in a flock of Chinese local-breed chickens infected with a subgroup J avian leukosis virus using suppression subtractive hybridization. AB - Avian leukosis virus subgroup J (ALV-J) is a new type of virus that mainly induces myeloid leukosis (ML) in chickens. To further elucidate the pathogenesis of ALV-J infection and tumor development, expression profiles from the bone marrow tissue of 15 infected and 18 non-infected birds from a local-breed poultry farm under naturally infected conditions, were analyzed by suppression subtractive hybridization. The birds were diagnosed as ML+ (or ML-) by specific ALV-J detection methods, involving serological tests for antigens and antibodies, and RT-PCR to detect viral RNA. A total of 59 partial gene sequences were revealed by differential screening of 496 forward and 384 reverse subtracted cDNA clones. Of these, 22 identified genes, including 8 up-regulated and 14 down regulated, were related to immune functions, these genes being, MHC B-G antigen, translationally-controlled tumor protein (TPT1/TPTC), transferrin and ferritin, hemoglobin and Carbonic anhydrase. Four of the down-regulated genes were selected for further analysis, in view of their predicted roles in infection and immunity by real-time qRT-PCR, using RNA collected from the same birds as those used for SSH. The four genes were expressed at significantly lower levels (p < 0.001) in ALV-J infected birds than in non-infected ones. PMID- 21637604 TI - Microsatellite analysis to estimate genetic relationships among five bulgarian sheep breeds. AB - Herein, genetic relationships among five breeds of Bulgarian sheep were estimated using microsatellite markers. The total number of alleles identified was 226 at the 16 loci examined. D(A) distance values were used for phylogenetic tree construction with the UPGMA algorithm. The two Tsigai and two Maritza populations were found to be geneticallvery closely related to each other y (0.198, and 0.258 respectively). The Pleven Black Head population was distinct from the other four. These results could be useful for preserving genes in these breeds, thereby ensuring their preservation in Bulgaria. PMID- 21637605 TI - Karyological study of Amphisbaena ridleyi (Squamata, Amphisbaenidae), an endemic species of the Archipelago of Fernando de Noronha, Pernambuco, Brazil. AB - The karyotype of Amphisbaena ridleyi, an endemic species of the archipelago of Fernando de Noronha, in State of Pernambuco, Brazil, is described after conventional staining, Ag-NOR impregnation and fluorescence in situ hybridization (FISH) with a telomeric probe. The diploid number is 46, with nine pairs of macrochromosomes (three metacentrics, four subtelocentrics and two acrocentrics) and 14 pairs of microchromosomes. The Ag-NOR is located in the telomeric region of the long arm of metacentric chromosome 2 and FISH revealed signals only in the telomeric region of all chromosomes. Further cytogenetic data on other amphisbaenians as well as a robust phylogenetic hypothesis of this clade is needed in order to understand the evolutionary changes on amphisbaenian karyotypes. PMID- 21637606 TI - Evaluation of genetic variability in the collared peccary Pecari tajacu and the white-lipped peccary Tayassu pecari by microsatellite markers. AB - In this study, the microsatellite technique was used to evaluate the genetic variability in populations of collared and white-lipped peccaries kept in captivity. Six primers developed for domestic pigs were used and amplified in both species. They revealed the presence of five polymorphic loci and one monomorphic locus. The polymorphic loci included 4 of the 16 alleles in collared peccaries, and 3 of the 10 alleles in the white-lipped peccaries. Polymorphic information content (PIC) in both species and all the loci was highly informative. The probability of paternity exclusion (PEC), if one of the parents is known, was almost as high in white-lipped peccaries (95.53%) as in the collared (99,48%). The Fst values for collared (0.042) and white-lipped (0.1387) peccaries showed that both populations are not structured. The Fis values for all loci, except ACTG2 in white-lipped peccaries (-0.0275) and in both species (0.1985 to 0.9284 in collared peccaries and 0.3621 to 0.4754 in the white lipped), revealed a high level of homozygosis, probably caused by inbreeding. Data on heterologous amplification and genetic variability in collared and white lipped peccaries are presented for the first time. PMID- 21637607 TI - Association between MUC1 gene polymorphism and expected progeny differences in Nelore cattle (Bos primigenius indicus). AB - MUC1 is a heavily glycosylated mammalian transmembrane protein expressed by mucosal secretory tissues for both protection against microbial infection and lubrication. An important characteristic of MUC1 is its variable number of tandem repeats (VNTR) containing several sites for O-glycosylation. VNTR length has been associated with many human diseases and with certain economically important traits in domestic ruminants. The aim of the present study was to correlate the length of MUC1 gene VNTR with expected progeny differences (EPDs) obtained for growth, fertility and carcass traits. Five alleles were identified, with alleles containing short VNTRs being more frequent than those with long, thereby demonstrating that Brazilian Nelore cattle are characterized by high frequencies in short MUC1 VNTRs. Statistical analyses revealed there to be no significant association between VNTR length and EPDs for weight at 120 days (W(120) ), scrotal circumference at 365 (SC (365) ) and 450 (SC (450) ) days, age at first calving (AFC), and rib eye area (REA). PMID- 21637608 TI - Genetic parameters for milk, fat and protein yields in Murrah buffaloes (Bubalus bubalis Artiodactyla, Bovidae). AB - The objective of the present study was to estimate genetic parameters for test day milk, fat and protein yields and 305-day-yields in Murrah buffaloes. 4,757 complete lactations of Murrah buffaloes were analyzed. Co-variance components were estimated by the restricted maximum likelihood method. The models included additive direct genetic and permanent environmental effects as random effects, and the fixed effects of contemporary group, milking number and age of the cow at calving as linear and quadratic covariables. Contemporary groups were defined by herd-year-month of test for test-day yields and by herd-year-season of calving for 305-day yields. The heritability estimates obtained by two-trait analysis ranged from 0.15 to 0.24 for milk, 0.16 to 0.23 for protein and 0.13 to 0.22 for fat, yields. Genetic and phenotypic correlations were all positive. The observed population additive genetic variation indicated that selection might be an effective tool in changing population means in milk, fat and protein yields. PMID- 21637609 TI - High levels of genetic differentiation and selfing in the Brazilian cerrado fruit tree Dipteryx alata Vog. (Fabaceae). AB - Dipteryx alata is a native fruit tree species of the cerrado (Brazilian savanna) that has great economic potential because of its multiple uses. Knowledge of how the genetic variability of this species is organized within and among populations would be useful for genetic conservation and breeding programs. We used nine simple sequence repeat (SSR) primers developed for Dipteryx odorata to evaluate the genetic structure of three populations of D. alata located in central Brazil based on a leaf sample analysis from 101 adults. The outcrossing rate was evaluated using 300 open-pollinated offspring from 25 seed-trees. Pollen dispersal was measured by parentage analysis. We used spatial genetic structure (SGS) to test the minimal distance for harvesting seeds in conservation and breeding programs. Our data indicate that the populations studied had a high degree of genetic diversity and population structure, as suggested by the high level of divergence among populations . The estimated outcrossing rate suggested a mixed mating system, and the intrapopulation fixation index was influenced by SGS. We conclude that seed harvesting for genetic conservation and breeding programs requires a minimum distance between trees of 196 m to avoid collecting seeds from related seed-trees. PMID- 21637610 TI - Clonal diversity and conservation genetics of the medicinal plant Carapichea ipecacuanha (Rubiaceae). AB - The roots of the understorey shrub Carapichea ipecacuanha (ipecac) have medicinal properties, and the uprooting of wild plants has supplied most of the world demand for this species. Although under severe population decline, C. ipecacuanha lacks legal protection. In the wild, the aerial stems of ipecac clump together to form clusters with well-defined borders. Cluster size may range from several to hundreds of aerial stems. To investigate the extent of clonality among aerial stems in ipecac clusters, we sampled 50 wild clusters (a total of 291 aerial stems) and screened them with 89 inter-simple sequence repeat (ISSR) markers. The 291 aerial stems were grouped into 42 putative clones. The clonal groups generally consisted of aerial stems from the same cluster, and there was little or no genetic differentiation among aerial stems at the cluster level. These findings suggest that strategies designed to conserve ipecac in situ should not rely upon census data, which are based on the number of aerial stems per cluster and the number of clusters per population, because such data greatly underestimate the species effective population size and genetic diversity. Our results also indicate that this species needs protection at a federal level. PMID- 21637611 TI - Chromatin differentiation between Theobroma cacao L. and T. grandiflorum Schum. AB - A comparative analysis of mitotic chromosomes of Theobroma cacao (cacao) and T. grandiflorum (cupuacu) was performed aiming to identify cytological differences between the two most important species of this genus. Both species have symmetric karyotypes, with 2n = 20 metacentric chromosomes ranging in size from 2.00 to 1.19 MUm (cacao) and from 2.21 to 1.15 MUm (cupuacu). The interphase nuclei of both species were of the arreticulate type, displaying up to 20 chromocentres, which were more regularly shaped in cacao than in cupuacu. Prophase chromosomes of both species were more condensed in the proximal region, sometimes including the whole short arm. Both species exhibited only one pair of terminal heterochromatic bands, positively stained with chromomycin A (3) , which co localized with the single 45S rDNA site. Each karyotype displayed a single 5S rDNA site in the proximal region of another chromosome pair. Heterochromatic bands were also observed on the centromeric/pericentromeric regions of all 20 chromosomes of cacao after C-banding followed by Giemsa or DAPI staining, whereas in cupuacu they were never detected. These data suggest that the chromosomes of both species have been largely conserved and their pericentromeric chromatin is the only citologically differentiated region. PMID- 21637612 TI - The use and limits of ITS data in the analysis of intraspecific variation in Passiflora L. (Passifloraceae). AB - The discovery and characterization of informative intraspecific genetic markers is fundamental for evolutionary and conservation genetics studies. Here, we used nuclear ribosomal ITS sequences to access intraspecific genetic diversity in 23 species of the genus Passiflora L. Some degree of variation was detected in 21 of these. The Passiflora and Decaloba (DC.) Rchb. subgenera showed significant differences in the sizes of the two ITS regions and in GC content, which can be related to reproductive characteristics of species in these subgenera. Furthermore, clear geographical patterns in the spatial distribution of sequence types were identified in six species. The results indicate that ITS may be a useful tool for the evaluation of intraspecific genetic variation in Passiflora. PMID- 21637613 TI - Genetic diversity analysis in the section Caulorrhizae (genus Arachis) using microsatellite markers. AB - Diversity in 26 microsatellite loci from section Caulorrhizae germplasm was evaluated by using 33 accessions of A. pintoi Krapov. & W.C. Gregory and ten accessions of Arachis repens Handro. Twenty loci proved to be polymorphic and a total of 196 alleles were detected with an average of 9.8 alleles per locus. The variability found in those loci was greater than the variability found using morphological characters, seed storage proteins and RAPD markers previously used in this germplasm. The high potential of these markers to detect species-specific alleles and discriminate among accessions was demonstrated. The set of microsatellite primer pairs developed by our group for A. pintoi are useful molecular tools for evaluating Section Caulorrhizae germplasm, as well as that of species belonging to other Arachis sections. PMID- 21637614 TI - Comparative karyotypic analysis in the Alstroemeria hookeri Lodd. (Alstroemeriaceae) complex sensu Bayer (1987). AB - Alstroemeria L. (Alstroemeriaceae) is an American genus of monocots with two principal distribution centers in Chile and Brazil. In Chile, it is represented by about 32 species, most of them in central Chile, an area known for its high level of endemism. The "complex" Alstroemeriahookeri is endemic to Chile, where it is distributed from the Coquimbo to the Bio-Bio Region. We analyzed the karyotypes of 36 populations of this complex along its natural distribution. Ten metaphases per population were used for chromosome measurements. All analyzed subspecies presented a well defined asymmetric karyotype. The populations of A. hookeri subsp. hookeri collected in the coastal range of the Bio-Bio Region and the populations from the Central Valley of this Region (Pangal del Laja) presented striking morphological differences in the karyotype, mainly on chromosome 3. The population of A. hookeri subsp. recumbens from Pichicuy showed a polymorphism on chromosome 7, which differed from the other analyzed populations of this subspecies. Phenetic analysis suggested that A. hookeri subsp. cummingiana, which showed a more symmetrical karyotype and did not grow in sandy soil, should be alocated to A. cummingiana rather than considered as part of the hookeri complex. PMID- 21637615 TI - Epistasis and genotype-by-environment interaction of grain protein content in durum wheat. AB - Parental, F(1) , F (2) , BC (1) and BC (2) generations of four crosses involving four cultivars of durum wheat (Triticum durum Desf.) were evaluated at two sites in Tunisia. A three-parameter model was found inadequate for all cases except crosses Chili x Cocorit 71 at site Sidi Thabet and Inrat 69 x Karim at both sites. In most cases a digenic epistatic model was sufficient to explain variation in generation means. Dominance effects (h) and additive x additive epistasis (i) (when significant) were more important than additive (d) effects and other epistatic components. Considering the genotype-by-environment interaction, the non-interactive model (m, d, h, e) was found adequate. Additive variance was higher than environmental variance in three crosses at both sites. The estimated values of narrow-sense heritability were dependent upon the cross and the sites and were 0%-85%. The results indicate that appropriate choice of environment and selection in later generations would increase grain protein content in durum wheat. PMID- 21637616 TI - Genetic characterization of 12 heterologous microsatellite markers for the giant tropical tree Cariniana legalis (Lecythidaceae). AB - Twelve microsatellite loci previously developed in the tropical tree Cariniana estrellensis were genetically characterized in Cariniana legalis. Polymorphisms were assessed in 28 C. legalis individuals found between the Pardo and Mogi-Guacu River basins in the state of Sao Paulo, Brazil. Of the 12 loci, 10 were polymorphic and exhibited Mendelian inheritance. The allelic richness at each locus ranged from 2-11, with an average of 7 alleles per locus, and the expected heterozygosity ranged from 0.07-0.88. These loci showed a high probability of paternity exclusion. The characteristics of these heterologous microsatellite markers indicate that they are suitable tools for investigating questions concerning population genetics in C. legalis. PMID- 21637617 TI - Mutations in retrotransposon AtCOPIA4 compromises resistance to Hyaloperonospora parasitica in Arabidopsis thaliana. AB - Retrotransposons (RTEs) are a principal component of most eukaryotic genomes, representing 50%-80% of some grass genomes. RTE sequences have been shown to be preferentially present in disease resistance gene clusters in plants. Arabidopsis thaliana has over 1,600 annotated RTE sequences and 56 of these appear to be expressed because of the exact expressed sequence tag (EST) matches and the presence of intact open reading frames. Of the 22 represented in the Affymetrix ATH1 array, AtCOPIA4 was found to be expressed at a higher level than all other RTEs across different developmental stages. Since AtCOPIA4 is located in the RPP5 gene cluster and is adjacent to RPP4 which confers resistance to the downy mildew oomycete Hyaloperonospora parasitica isolate EMWA1, we evaluated AtCOPIA4 mutants for resistance to this pathogen. T-DNA insertional and antisense knockout of AtCOPIA4 was found to reduce the resistance of wild type plants by 2-4 folds. Our results suggest that retrotransposon can be exapted to participate in plant defense response. PMID- 21637618 TI - Unusual intron in the second exon of a Type III polyketide synthase gene of Alpinia calcarata Rosc. AB - Plant phenolic compounds form a valuable resource of secondary metabolites having a broad spectrum of biological activities. Type III polyketide synthases play a key role in the formation of basic structural skeleton of the phenolic compounds. As a group of medicinal plants, PKSs with novel features are expected in the genome of Zingiberaceae. The genomic exploration of PKS in Alpinia calcarata conducted in this study identified the presence of an unusual intron at the region forming the second exon of typical PKSs, forming a gateway information of distribution of novel PKSs in Zingiberaceae. PMID- 21637619 TI - Mutation in the xpsD gene of Xanthomonas axonopodis pv. citri affects cellulose degradation and virulence. AB - The Gram-negative bacterium Xanthomonas axonopodis pv. citri, the causal agent of citrus canker, is a major threat to the citrus industry worldwide. Although this is a leaf spot pathogen, it bears genes highly related to degradation of plant cell walls, which are typically found in plant pathogens that cause symptoms of tissue maceration. Little is known on Xac capacity to cause disease and hydrolyze cellulose. We investigated the contribution of various open reading frames on degradation of a cellulose compound by means of a global mutational assay to selectively screen for a defect in carboxymethyl cellulase (CMCase) secretion in X. axonopodis pv. citri. Screening on CMC agar revealed one mutant clone defective in extracellular glycanase activity, out of nearly 3,000 clones. The insertion was located in the xpsD gene, a component of the type II secretion system (T2SS) showing an influence in the ability of Xac to colonize tissues and hydrolyze cellulose. In summary, these data show for the first time, that X. axonopodis pv. citri is capable of hydrolyzing cellulose in a T2SS-dependent process. Furthermore, it was demonstrated that the ability to degrade cellulose contributes to the infection process as a whole. PMID- 21637620 TI - Investigations on DNA damage and frequency of micronuclei in occupational exposure to electromagnetic fields (EMFs) emitted from video display terminals (VDTs). AB - The potential effect of electromagnetic fields (EMFs) emitted from video display terminals (VDTs) to elicit biological response is a major concern for the public. The software professionals are subjected to cumulative EMFs in their occupational environments. This study was undertaken to evaluate DNA damage and incidences of micronuclei in such professionals. To the best of our knowledge, the present study is the first attempt to carry out cytogenetic investigations on assessing bioeffects in personal computer users. The study subjects (n = 138) included software professionals using VDTs for more than 2 years with age, gender, socioeconomic status matched controls (n = 151). DNA damage and frequency of micronuclei were evaluated using alkaline comet assay and cytochalasin blocked micronucleus assay respectively. Overall DNA damage and incidence of micronuclei showed no significant differences between the exposed and control subjects. With exposure characteristics, such as total duration (years) and frequency of use (minutes/day) sub-groups were assessed for such parameters. Although cumulative frequency of use showed no significant changes in the DNA integrity of the classified sub-groups, the long-term users (> 10 years) showed higher induction of DNA damage and increased frequency of micronuclei and micro nucleated cells. PMID- 21637621 TI - Alterations in gene expression profiles correlated with cisplatin cytotoxicity in the glioma U343 cell line. AB - Gliomas are the most common tumors in the central nervous system, the average survival time of patients with glioblastoma multiforme being about 1 year from diagnosis, in spite of harsh therapy. Aiming to study the transcriptional profiles displayed by glioma cells undergoing cisplatin treatment, gene expression analysis was performed by the cDNA microarray method. Cell survival and apoptosis induction following treatment were also evaluated. Drug concentrations of 12.5 to 300 MUM caused a pronounced reduction in cell survival rates five days after treatment, whereas concentrations higher than 25 MUM were effective in reducing the survival rates to ~1%. However, the maximum apoptosis frequency was 20.4% for 25 MUM cisplatin in cells analyzed at 72 h, indicating that apoptosis is not the only kind of cell death induced by cisplatin. An analysis of gene expression revealed 67 significantly (FDR < 0.05) modulated genes: 29 of which down- and 38 up-regulated. These genes belong to several classes (metabolism, protein localization, cell proliferation, apoptosis, adhesion, stress response, cell cycle and DNA repair) that may represent several affected cell processes under the influence of cisplatin treatment. The expression pattern of three genes (RHOA, LIMK2 and TIMP2) was confirmed by the real time PCR method. PMID- 21637622 TI - Genotoxic evaluation of an industrial effluent from an oil refinery using plant and animal bioassays. AB - Polycyclic aromatic hydrocarbons (PAHs) are genotoxic chemicals commonly found in effluents from oil refineries. Bioassays using plants and cells cultures can be employed for assessing environmental safety and potential genotoxicity. In this study, the genotoxic potential of an oil refinery effluent was analyzed by means of micronucleus (MN) testing of Alium cepa, which revealed no effect after 24 h of treatment. On the other hand, primary lesions in the DNA of rat (Rattus norvegicus) hepatoma cells (HTC) were observed through comet assaying after only 2 h of exposure. On considering the capacity to detect DNA damage of a different nature and of these cells to metabolize xenobiotics, we suggest the association of the two bioassays with these cell types, plant (Allium cepa) and mammal (HTC) cells, for more accurately assessing genotoxicity in environmental samples. PMID- 21637623 TI - Determination of the antimutagenicity of an aqueous extract of Rhizophora mangle L. (Rhizophoraceae), using in vivo and in vitro test systems. AB - An aqueous extract of Rhizophora mangle L. bark is used as raw material in pottery making in the State of Espirito Santo, Brazil. This extract presents large quantities of tannins, compounds possessing antioxidant properties. Tannin antioxidant activity, as a plant chemical defense mechanism in the process of stabilizing free radicals, has been an incentive to studies on anti-mutagenicity. The present work aimed to evaluate possible antimutagenic activity of a R. mangle aqueous extract, using the Allium cepa test-system and micronuclear (MN) assay with blockage of cytokinesis in Chinese hamster ovary cells (CHO-K1). The Allium cepa test-system indicated antimutagenic activity against the damage induced by the mutagenic agent methyl methanesulfonate. A reduction in both MN cell frequency and chromosome breaks occurred in both the pre and post-treatment protocols. The MN testing of CHO-K1 cells revealed anti-mutagenic activity of the R. mangle extract against methyl methanesulfonate and doxorubicin in pre, simultaneous and post-treatment protocols. These results suggest the presence of phyto-constituents in the extract presenting demutagenic and bio-antimutagenic activities. Since the chemical constitution of Rhizophora mangle species presents elevated tannin content, it is highly probable that these compounds are the antimutagenic promoters themselves. PMID- 21637624 TI - Protective effect of Hemidesmus indicus R.Br. root extract against cisplatin induced cytogenetic damage in mouse bone marrow cells. AB - The aqueous extract of Hemidesmus indicus roots was investigated for its in vivo antigenotoxic effect against cisplatin-induced cytogenetic damage. Swiss albino mice were administered with various doses of the extract either singly (50, 100 and 200 mg/kg body weight) or as split doses (10, 20 and 40 mg/kg bw/day) for five consecutive days by oral gavage. As endpoints, chromosome aberrations, micronuclei in polychromatic erythrocytes, mitotic index and PCE/NCE ratio were estimated. The extract protected the bone marrow cells from cisplatin-induced genotoxicity in an inverse dose-dependent manner. However, the extract was cytotoxic at all doses. But, under split dose regime it conferred a higher level of genoprotection and was not cytotoxic at the lower two doses. The presence of saponins, tannins, phenols, terpenoids, flavonoids and coumarins in the crude extract could explain these effects. PMID- 21637625 TI - Mitochondrial genome nucleotide substitution pattern between domesticated silkmoth, Bombyx mori, and its wild ancestors, Chinese Bombyx mandarina and Japanese Bombyx mandarina. AB - Bombyx mori and Bombyx mandarina are morphologically and physiologically similar. In this study, we compared the nucleotide variations in the complete mitochondrial (mt) genomes between the domesticated silkmoth, B. mori, and its wild ancestors, Chinese B. mandarina (ChBm) and Japanese B. mandarina (JaBm). The sequence divergence and transition mutation ratio between B. mori and ChBm are significantly smaller than those observed between B. mori and JaBm. The preference of transition by DNA strands between B. mori and ChBm is consistent with that between B. mori and JaBm, however, the regional variation in nucleotide substitution rate shows a different feature. These results suggest that the ChBm mt genome is not undergoing the same evolutionary process as JaBm, providing evidence for selection on mtDNA. Moreover, investigation of the nucleotide sequence divergence in the A+T-rich region of Bombyx mt genomes also provides evidence for the assumption that the A+T-rich region might not be the fastest evolving region of the mtDNA of insects. PMID- 21637626 TI - Novel sequence variations in LAMA2 andSGCG genes modulating cis-acting regulatory elements and RNA secondary structure. AB - In this study, we detected new sequence variations in LAMA2 and SGCG genes in 5 ethnic populations, and analysed their effect on enhancer composition and mRNA structure. PCR amplification and DNA sequencing were performed and followed by bioinformatics analyses using ESEfinder as well as MFOLD software. We found 3 novel sequence variations in the LAMA2 (c.3174+22_23insAT and c.6085 +12delA) and SGCG (c. (*) 102A/C) genes. These variations were present in 210 tested healthy controls from Tunisian, Moroccan, Algerian, Lebanese and French populations suggesting that they represent novel polymorphisms within LAMA2 and SGCG genes sequences. ESEfinder showed that the c. (*) 102A/C substitution created a new exon splicing enhancer in the 3'UTR of SGCG genes, whereas the c.6085 +12delA deletion was situated in the base pairing region between LAMA2 mRNA and the U1snRNA spliceosomal components. The RNA structure analyses showed that both variations modulated RNA secondary structure. Our results are suggestive of correlations between mRNA folding and the recruitment of spliceosomal components mediating splicing, including SR proteins. The contribution of common sequence variations to mRNA structural and functional diversity will contribute to a better study of gene expression. PMID- 21637627 TI - Use of direct and iterative solvers for estimation of SNP effects in genome-wide selection. AB - The aim of this study was to compare iterative and direct solvers for estimation of marker effects in genomic selection. One iterative and two direct methods were used: Gauss-Seidel with Residual Update, Cholesky Decomposition and Gentleman Givens rotations. For resembling different scenarios with respect to number of markers and of genotyped animals, a simulated data set divided into 25 subsets was used. Number of markers ranged from 1,200 to 5,925 and number of animals ranged from 1,200 to 5,865. Methods were also applied to real data comprising 3081 individuals genotyped for 45181 SNPs. Results from simulated data showed that the iterative solver was substantially faster than direct methods for larger numbers of markers. Use of a direct solver may allow for computing (co)variances of SNP effects. When applied to real data, performance of the iterative method varied substantially, depending on the level of ill-conditioning of the coefficient matrix. From results with real data, Gentleman-Givens rotations would be the method of choice in this particular application as it provided an exact solution within a fairly reasonable time frame (less than two hours). It would indeed be the preferred method whenever computer resources allow its use. PMID- 21637628 TI - Other research interests regarding biomarkers in the population with alcohol dependency. PMID- 21637629 TI - Personality disorders: a nation-based perspective on prevalence. AB - To date, five major studies have examined the prevalence and type of personality disorders in community samples in the United States. According to the majority of studies, the overall prevalence of Axis II disorders in the general population is consistently around 10 percent. According to the most recent study, obsessive compulsive personality disorder is the most frequent Axis II disorder in community samples in the United States, followed by narcissistic and borderline personality disorders. In contrast to studies in the United States, community prevalence rates of personality disorders in other countries show moderately wide variation, from 6.1 to 13.4 percent; yet, the averaging of these two low/high percentages results in 9.7 percent. The most common type of personality pathology in a given country varies, and this variance may be accounted for in a number of relevant ways. PMID- 21637630 TI - The use of research measures in adult clinical practice. AB - Many psychopathology research assessment tools can be used easily and productively in clinical practice. We conducted a workshop in 2009 and 2010 at the American Psychiatric Association annual meeting designed to bring clinicians some commonly used adult research measures with broad applicability to a variety of conditions. This article reviews what was most helpful to the practicing clinicians at the workshop. PMID- 21637631 TI - Reflexes in psychiatry. AB - Psychiatric patients often do not cooperate fully with the neurologic examination. Reflexes virtually bypass patient effort and are difficult to consciously determine. This article reviews muscle stretch (deep tendon) reflexes, and pathological reflexes including the extensor plantar (Babinski) and primitive release reflexes. Topics include findings in common psychiatric and neurologic conditions and methods for eliciting these signs. PMID- 21637632 TI - Narcolepsy presenting as schizophrenia: a literature review and two case reports. AB - Schizophrenia is a psychiatric disorder that causes significant disability and morbidity. Narcolepsy is a disorder, less prevalent than schizophrenia, but a disorder in which symptoms overlap with schizophrenia. This overlap in symptoms can cause narcolepsy to be confused with schizophrenia. The differences and similarities between narcolepsy and schizophrenia are discussed in the context of two cases. The first case describes an adolescent and the second case describes a refractory case of narcolepsy that did not respond to stimulants. Both cases were previously diagnosed as schizophrenia and did not respond to trials of antipsychotics and other psychotropics. The patients were reevaluated and referred to sleep testing at our facility for clinical suspicion of narcolepsy. Both patients underwent polysomnography with subsequent multiple sleep latency testing. The sleep testing results and multiple sleep latency testing criteria for narcolepsy are also discussed. The patients were treated for narcolepsy resulting in remission of the psychotic symptoms with significant behavioral improvement. We recommend that psychiatrists consider narcolepsy in the differential diagnosis when faced with refractory psychosis. PMID- 21637633 TI - Are clozapine blood dyscrasias associated with concomitant medications? AB - Clozapine is an atypical antipsychotic agent used for refractory schizophrenia. It has a relatively low affinity for D2 receptors and thus is associated with a lower incidence of extrapyramidal side effects when compared with typical antipsychotics. Clozapine as monotherapy can induce a rare, but serious, blood dyscrasia called agranulocytosis; however, some concomitant medications may contribute to the risk. Examples of these medications are mood-stabilizing antiepileptic drugs, such as carbamazepine, and sulfonamide antibiotics, such as sulfamethoxazole. There were no studies at the writing of this article examining the effect of concomitant medications on clozapine blood dyscrasias, and few published reports describing enhanced bone marrow suppression in those taking clozapine. The primary objective of this study was to evaluate the effect of concomitant medications used in a state psychiatric hospital on clozapine-induced blood dyscrasias. This was a retrospective record review of adverse drug reactions reported at an adult inpatient state psychiatric center. The records for a pilot sample of 26 patients with reported clozapine-related adverse drug reactions between January 1, 2007, and June 30, 2009, were reviewed. Fundamental to this study were reported adverse drug reactions defined as 1) substantial drops in white blood cell or absolute neutrophil count (a substantial drop in white blood cell is >3,000 or absolute neutrophil count is >1,500 over a 3-week period); 2) mild leukopenia/granulocytopenia; and 3) moderate-severe leukopenia/granulocytopenia. Concomitant medications were examined for contributions to an increased potential for clozapine-induced blood dyscrasias. Other data collected included demographic information (age, gender, ethnicity), medical and psychiatric diagnoses, dose and duration of medications, and changes in medications. Medications that had a statistically significant impact on the incidence of clozapine-induced blood dyscrasias are reported in this article, as well as the possible duration of medication use prior to induction of an adverse drug reaction. PMID- 21637634 TI - The social and treatment consequences of a shared delusional disorder in a homeless family. AB - OBJECTIVE: Delusional disorder is defined as a fixed, false belief that is held by a person despite evidence to the contrary. Shared psychotic disorder, also known as folie a' deux, psychosis by association and induced psychotic disorder, is an uncommon and unique psychiatric disorder. It is even more unusual when it occurs within families (folie a' famille). CASE PRESENTATION: This case report describes the occurrence of a shared delusion within a family consisting of an adults on and two elderly parents. The shared delusion, which was the belief that a large financial settlement was a waiting to be disbursed to the family members by the local law enforcement agency, contributed to their state of homelessness and rejection of all offers of assistance from service providers. CONCLUSION: The impact of this shared psychotic disorder contributed to the family's state of extreme poverty and homelessness, which, as a consequence, greatly impeded the initiation of evidence-based therapeutic interventions. PMID- 21637635 TI - Hereditary breast and ovarian cancer: new genes, new treatments, new concepts. AB - BACKGROUND: Every year, 60,000 women in Germany are found to have breast cancer, and 9000 to have ovarian cancer. Familial clustering of carcinoma is seen in about 20% of cases. METHODS: We selectively review relevant articles published up to December 2010 that were retrieved by a search in PubMed, and we also discuss findings from the experience of the German Consortium for Hereditary Breast and Ovarian Cancer. RESULTS: High risk is conferred by the highly penetrant BRCA1 and BRCA2 genes as well as by other genes such as RAD51C. Genes for breast cancer that were originally designated as moderately penetrant display higher penetrance than previously thought in families with a hereditary predisposition. The role these genes play in DNA repair is thought to explain why tumors associated with them are sensitive to platin derivatives and PARP inhibitors. In carriers of BRCA1 and BRCA2, prophylactic bilateral mastectomy and adnexectomy significantly lowers the incidence of breast and ovarian cancer. Moreover, prophylactic adnexectomy also lowers the breast-and-ovarian-cancer-specific mortality, as well as the overall mortality. If a woman bearing a mutation develops cancer in one breast, her risk of developing cancer in the other breast depends on the particular gene that is mutated and on her age at the onset of disease. CONCLUSION: About half of all monogenically determined carcinomas of the breast and ovary are due to a mutation in one or the other of the highly penetrant BRCA genes (BRCA1 and BRCA2). Women carrying a mutated gene have an 80% to 90% chance of developing breast cancer and a 20% to 50% chance of developing ovarian cancer. Other predisposing genes for breast and ovarian cancer have been identified. Clinicians should develop and implement evidence-based treatments on the basis of these new findings. PMID- 21637638 TI - Deficits in the management of patients with adrenocortical carcinoma in Germany. Statements cannot be substantiated. PMID- 21637636 TI - Percutaneous vertebroplasty in vertebral compression fractures of benign or malignant origin: a prospective study of 1188 patients with follow-up of 12 months. AB - BACKGROUND: Vertebral body fractures are a source of high costs for the health care system and will continue to be one as the population ages. Cost-effective treatment is thus all the more important. In this study, we evaluated patients' quality of life during the first 12 months after they had undergone percutaneous vertebroplasty for vertebral body fractures which were refractory to conservative treatment. Our analysis took the causes of the fractures into account. METHODS: Pain, mobility, and need for analgesics were assessed prospectively on verbal rating scales one day before and one day after vertebroplasty, as well as over a follow-up period of up to 12 months. The same examiner interviewed each patient at all time points to obtain this information. RESULTS: 1188 patients underwent vertebroplasty for 1980 vertebral body fractures; the most common etiology was osteoporosis (75%). There was statistically relevant improvement in all three of the variables studied from the day before the procedure to the last follow-up, regardless of the cause of fracture (p<0.01). Most of the clinical benefit was already evident on the day after the procedure. Patients with fractures due to osteoporosis experienced further statistically relevant improvement by 6 months after treatment. CONCLUSION: Percutaneous vertebroplasty immediately relieves the pain of vertebral body fractures, improves patients' mobility, and lowers their consumption of analgesics. There can be further clinical improvement up to 6 months after the procedure, particularly in patients with fractures due to osteoporosis. As osteoporosis is the most common cause of vertebral body fractures, this patient group is important not just clinically, but economically as well. PMID- 21637639 TI - Genetic signatures of parental contribution in black and white populations in Brazil. AB - Two hundred and three individuals classified as white were tested for 11 single nucleotide polymorphisms plus two insertion/deletions in their Y-chromosomes. A subset of these individuals (n = 172) was also screened for sequences in the first hypervariable segment of their mitochondrial DNA (mtDNA). In addition, complementary studies were done for 11 of the 13 markers indicated above in 54 of 107 black subjects previously investigated in this southern Brazilian population. The prevalence of Y-chromosome haplogroups among whites was similar to that found in the Azores (Portugal) or Spain, but not to that of other European countries. About half of the European or African mtDNA haplogroups of these individuals were related to their places of origin, but not their Amerindian counterparts. Persons classified in these two categories of skin color and related morphological traits showed distinct genomic ancestries through the country. These findings emphasize the need to consider in Brazil, despite some general trends, a notable heterogeneity in the pattern of admixture dynamics within and between populations/groups. PMID- 21637640 TI - Analysis of the CCR5 gene coding region diversity in five South American populations reveals two new non-synonymous alleles in Amerindians and high CCR5*D32 frequency in Euro-Brazilians. AB - The CC chemokine receptor 5 (CCR5) molecule is an important co-receptor for HIV. The effect of the CCR5*D32 allele in susceptibility to HIV infection and AIDS disease is well known. Other alleles than CCR5*D32 have not been analysed before, neither in Amerindians nor in the majority of the populations all over the world. We investigated the distribution of the CCR5 coding region alleles in South Brazil and noticed a high CCR5*D32 frequency in the Euro-Brazilian population of the Parana State (9.3%), which is the highest thus far reported for Latin America. The D32 frequency is even higher among the Euro-Brazilian Mennonites (14.2%). This allele is uncommon in Afro-Brazilians (2.0%), rare in the Guarani Amerindians (0.4%) and absent in the Kaingang Amerindians and the Oriental Brazilians. R223Q is common in the Oriental-Brazilians (7.7%) and R60S in the Afro-Brazilians (5.0%). A29S and L55Q present an impaired response to beta chemokines and occurred in Afro- and Euro-Brazilians with cumulative frequencies of 4.4% and 2.7%, respectively. Two new non-synonymous alleles were found in Amerindians: C323F (g.3729G > T) in Guarani (1.4%) and Y68C (g.2964A > G) in Kaingang (10.3%). The functional characteristics of these alleles should be defined and considered in epidemiological investigations about HIV-1 infection and AIDS incidence in Amerindian populations. PMID- 21637641 TI - Simultaneous quantitative assessment of circulating cell-free mitochondrial and nuclear DNA by multiplex real-time PCR. AB - Quantification of circulating nucleic acids in plasma and serum could be used as a non-invasive diagnostic tool for monitoring a wide variety of diseases and conditions. We describe here a rapid, simple and accurate multiplex real-time PCR method for direct synchronized analysis of circulating cell-free (ccf) mitochondrial (mtDNA) and nuclear (nDNA) DNA in plasma and serum samples. The method is based on one-step multiplex real-time PCR using a FAM-labeled MGB probe and primers to amplify the mtDNA sequence of the ATP 8 gene, and a VIC-labeled MGB probe and primers to amplify the nDNA sequence of the glycerinaldehyde-3 phosphate-dehydrogenase (GAPDH) gene, in plasma and serum samples simultaneously. The efficiencies of the multiplex assays were measured in serial dilutions. Based on the simulation of the PCR reaction kinetics, the relative quantities of ccf mtDNA were calculated using a very simple equation. Using our optimised real-time PCR conditions, close to 100% efficiency was obtained from the two assays. The two assays performed in the dilution series showed very good and reproducible correlation to each other. This optimised multiplex real-time PCR protocol can be widely used for synchronized quantification of mtDNA and nDNA in different samples, with a very high rate of efficiency. PMID- 21637642 TI - Estimating genomic instability mediated by Alu retroelements in breast cancer. AB - Alu-PCR is a relatively simple technique that can be used to investigate genomic instability in cancer. This technique allows identification of the loss, gain or amplification of gene sequences based on the analysis of segments between two Alu elements coupled with quantitative and qualitative analyses of the profiles obtained from tumor samples, surgical margins and blood. In this work, we used Alu-PCR to identify gene alterations in ten patients with invasive ductal breast cancer. Several deletions and insertions were identified, indicating genomic instability in the tumor and adjacent normal tissue. Although not associated with specific genes, the alterations, which involved chromosomal bands 1p36.23, 1q41, 11q14.3, 13q14.2, occurred in areas of well-known genomic instability in breast and other types of cancer. These results indicate the potential usefulness of Alu PCR in identifying altered gene sequences in breast cancer. However, caution is required in its application since the Alu primer can produce non-specific amplification. PMID- 21637643 TI - Polymorphism of the catechol-O-methyltransferase gene in Han Chinese patients with psoriasis vulgaris. AB - PSORIASIS VULGARIS IS DEFINED BY A SERIES OF LINKED CELLULAR CHANGES IN THE SKIN: hyperplasia of epidermal keratinocytes, vascular hyperplasia and ectasia, and infiltration of T lymphocytes, neutrophils and other types of leukocytes in the affected skin. Catechol-O-methyltransferase (COMT) 158 polymorphism can reduce the activity of the COMT enzyme that may trigger defective differentiation of keratinocytes in psoriasis. Immunocytes can degrade and inactivate catecholamines via monamine oxidase (MAO) and COMT in the cells. We hypothesized that the COMT 158G > A polymorphism was associated with the risk of psoriasis vulgaris in Han Chinese people. In a hospital-based case-control study, 524 patients with psoriasis vulgaris and 549 psoriasis-free controls were studied. COMT-158 G > A polymorphism was genotyped using the PCR sequence-specific primer (PCR-SSP) technique. We found no statistically significant association between the COMT-158 allele A and the risk of psoriasis vulgaris (p = 0.739 adjusted OR = 1.03; 95% CI = 0.81-1.31). This suggests that the COMT-158 G > A polymorphism may not contribute to the etiology of psoriasis vulgaris in the Han Chinese population. PMID- 21637644 TI - Chromosomal location of 18S and 5S rDNA sites in Triportheus fish species (Characiformes, Characidae). AB - The location of 18S and 5S rDNA sites was determined in eight species and populations of the fish genus Triportheus by using fluorescent in situ hybridization (FISH). The males and females of all species had 2n = 52 chromosomes and a ZZ/ZW sex chromosome system. A single 18S rDNA site that was roughly equivalent to an Ag-NOR was detected on the short arms of a submetacentric pair in nearly all species, and up to two additional sites were also observed in some species. In addition, another 18S rDNA cluster was identified in a distal region on the long arms of the W chromosome; this finding corroborated previous evidence that this cluster would be a shared feature amongst Triportheus species. In T. angulatus, a heterozygotic paracentric inversion involving the short arms of one homolog of a metacentric pair was associated with NORs. The 5S rDNA sites were located on the short arms of a single submetacentric chromosomal pair, close to the centromeres, except in T. auritus, which had up to ten 5S rDNA sites. The 18S and 5S rDNA sites were co localized and adjacent on the short arms of a chromosomal pair in two populations of T. nematurus. Although all Triportheus species have a similar karyotypic macrostructure, the results of this work show that in some species ribosomal genes may serve as species-specific markers when used in conjunction with other putatively synapomorphic features. PMID- 21637645 TI - Morphological variation in a secondary contact between divergent lineages of brown trout (Salmo trutta) from the Iberian Peninsula. AB - The aim of this study was to analyze the morphological variation of brown trout (Salmo trutta) in the Duero basin, an Atlantic river basin in the Iberian Peninsula, where a spatial segregation of two divergent lineages was previously reported, based on isozyme, microsatellite and mtDNA data. In these studies, two divergent pure regions (Pisuerga and Lower-course) and several hybrid populations between them were identified. Morphological variation was evaluated in 11 populations representative of the genetic differentiation previously observed in the Duero basin, using multivariate analysis on 12 morphometric and 4 meristic traits. A large differentiation between populations was observed (interpopulation component of variance: 41.8%), similar to that previously detected with allozymes and microsatellites. Morphometric differentiation was also reflected by the high classification success of pure and hybrid individuals to their respective populations, using multivariate discriminant functions (94.1% and 79.0%, respectively). All multivariate and clustering analyses performed demonstrated a strong differentiation between the pure regions. The hybrid populations, though showing large differentiation among them, evidenced an intermediate position between the pure samples. Head and body shape traits were the most discriminant among the morphometric characters, while pectoral rays and gillrakers were the most discriminant among the meristic traits. These results confirmed the high divergence of the brown trout from the Duero basin and suggest some traits on which selection could be acting to explain the spatial segregation observed. PMID- 21637646 TI - Biochemical comparison of two Hypostomus populations (Siluriformes, Loricariidae) from the Atlantico Stream of the upper Parana River basin, Brazil. AB - Two syntopic morphotypes of the genus Hypostomus - H. nigromaculatus and H. cf. nigromaculatus (Atlantico Stream, Parana State) - were compared through the allozyme electrophoresis technique. Twelve enzymatic systems (AAT, ADH, EST, GCDH, G3PDH, GPI, IDH, LDH, MDH, ME, PGM and SOD) were analyzed, attributing the score of 20 loci, with a total of 30 alleles. Six loci were diagnostic (Aat-2, Gcdh-1, Gpi-A, Idh-1, Ldh-A and Mdh-A), indicating the presence of interjacent reproductive isolation. The occurrence of few polymorphic loci acknowledge two morphotypes, with heterozygosity values He = 0.0291 for H. nigromaculatus and He = 0.0346 for H. cf. nigromaculatus. F(IS) statistics demonstrated fixation of the alleles in the two morphotypes. Genetic identity (I) and distance (D) of Nei (1978) values were I = 0.6515 and D = 0.4285. The data indicate that these two morphotypes from the Atlantico Stream belong to different species. PMID- 21637647 TI - Hidden heterochromatin: Characterization in the Rodentia species Cricetus cricetus, Peromyscus eremicus (Cricetidae) and Praomys tullbergi (Muridae). AB - The use of in situ restriction endonuclease (RE) (which cleaves DNA at specific sequences) digestion has proven to be a useful technique in improving the dissection of constitutive heterochromatin (CH), and in the understanding of the CH evolution in different genomes. In the present work we describe in detail the CH of the three Rodentia species, Cricetus cricetus, Peromyscus eremicus (family Cricetidae) and Praomys tullbergi (family Muridae) using a panel of seven REs followed by C-banding. Comparison of the amount, distribution and molecular nature of C-positive heterochromatin revealed molecular heterogeneity in the heterochromatin of the three species. The large number of subclasses of CH identified in Praomys tullbergi chromosomes indicated that the karyotype of this species is the more derived when compared with the other two genomes analyzed, probably originated by a great number of complex chromosomal rearrangements. The high level of sequence heterogeneity identified in the CH of the three genomes suggests the coexistence of different satellite DNA families, or variants of these families in these genomes. PMID- 21637648 TI - Polymorphisms on SSC15q21-q26 Containing QTL for reproduction in Swine and its association with litter size. AB - Several quantitative trait loci (QTL) for important reproductive traits (ovulation rate) have been identified on the porcine chromosome 15 (SSC15). To assist in the selection of positional candidate swine genes for these QTL on SSC15, twenty-one genes had already been assigned to SSC15 in a previous study in our lab, by using the radiation hybrid panel IMpRH. Further polymorphism studies were carried out on these positional candidate genes with four breeds of pigs (Duroc, Erhualian, Dahuabai and Landrace) harboring significant differences in reproduction traits. A total of nineteen polymorphisms were found in 21 genes. Among these, seven in six genes were used for association studies, whereby NRP2 polymorphism was found to be significantly (p < 0.05) associated with litter-size traits. NRP2 might be a candidate gene for pig-litter size based on its chromosome location (Du et al., 2006), significant association with litter-size traits and relationships with Sema and the VEGF super families. PMID- 21637649 TI - Bovine gene polymorphisms related to fat deposition and meat tenderness. AB - Leptin, thyroglobulin and diacylglycerol O-acyltransferase play important roles in fat metabolism. Fat deposition has an influence on meat quality and consumers' choice. The aim of this study was to determine allele and genotype frequencies of polymorphisms of the bovine genes, which encode leptin (LEP), thyroglobulin (TG) and diacylglycerol O-acyltransferase (DGAT1). A further objective was to establish the effects of these polymorphisms on meat characteristics. We genotyped 147 animals belonging to the Nelore (Bos indicus), Canchim (5/8 Bos taurus + 3/8 Bos indicus), Rubia Gallega X Nelore (1/2 Bos taurus + 1/2 Bos indicus), Brangus Three-way cross (9/16 Bos taurus + 7/16 Bos indicus) and Braunvieh Three-way cross (3/4 Bos taurus + 1/4 Bos indicus) breeds. Backfat thickness, total lipids, marbling score, ribeye area and shear force were fitted, using the General Linear Model (GLM) procedure of the SAS software. The least square means of genotypes and genetic groups were compared using Tukey's test. Allele frequencies vary among the genetic groups, depending on Bos indicus versus Bos taurus influence. The LEP polymorphism segregates in pure Bos indicus Nelore animals, which is a new finding. The T allele of TG is fixed in Nelore, and DGAT1 segregates in all groups, but the frequency of allele A is lower in Nelore animals. The results showed no association between the genotypes and traits studied, but a genetic group effect on these traits was found. So, the genetic background remains relevant for fat deposition and meat tenderness, but the gene markers developed for Bos taurus may be insufficient for Bos indicus. PMID- 21637650 TI - Cytogenetical studies in five Atlantic Anguilliformes fishes. AB - The order Anguilliformes comprises 15 families, 141 genera and 791 fish species. Eight families had at least one karyotyped species, with a prevalence of 2n = 38 chromosomes and high fundamental numbers (FN). The only exception to this pattern is the family Muraenidae, in which the eight species analyzed presented 2n = 42 chromosomes. Despite of the large number of Anguilliformes species, karyotypic reports are available for only a few representatives. In the present work, a species of Ophichthidae, Myrichthys ocellatus (2n = 38; 8m+14sm+10st+6a; FN = 70) and four species of Muraenidae, Enchelycore nigricans (2n = 42; 6m+8sm+12st+16a; FN = 68), Gymnothorax miliaris (2n = 42; 14m+18sm+10st; FN = 84), G. vicinus (2n = 42; 8m+6sm+28a; FN = 56) and Muraena pavonina (2n = 42; 6m+4sm+32a; FN = 52), collected along the Northeastern coast of Brazil and around the St Peter and St Paul Archipelago were analyzed. Typical large metacentric chromosomes were observed in all species. Conspicuous polymorphic heterochromatic regions were observed at the centromeres of most chromosomes and at single ribosomal sites. The data obtained for Ophichthidae corroborate the hypothesis of a karyotypic diversification mainly due to pericentric inversions and Robertsonian rearrangements, while the identification of constant chromosome numbers in Muraenidae (2n = 42) suggests a karyotype diversification through pericentric inversions and heterochromatin processes. PMID- 21637651 TI - Metaphase I orientation of Robertsonian trivalents in the water-hyacinth grasshopper, Cornops aquaticum (Acrididae, Orthoptera). AB - Trivalents resulting from polymorphic Robertsonian rearrangements must have a regular orientation in metaphase I if the polymorphisms are to be maintained. It has been argued that redistribution of proximal and interstitial chiasmata to more distal positions is necessary for a convergent orientation, the only one that produces viable gametes. Cornops aquaticum is a South-American grasshopper that lives and feeds on water-hyacinths, and has three polymorphic Robertsonian rearrangements in its southernmost distribution area in Central Argentina and Uruguay. The orientation of trivalents in metaphase I, the formation of abnormal spermatids and the frequency and position of chiasmata in the trivalents, was analysed in a polymorphic population of C. aquaticus. In this study we observed a correlation between the number of trivalents with the frequency of abnormal spermatids; additionally, the number of chiasmata, especially proximal and interstitial ones, was strongly correlated with the frequency of the linear orientation. Therefore we confirmed our previous assumption, based on other evidence, that the chiasmata redistribution in fusion carriers is essential to the maintenance of the polymorphisms. PMID- 21637652 TI - Genetic diversity of carotenoid-rich bananas evaluated by Diversity Arrays Technology (DArT). AB - The aim of this work was to evaluate the carotenoid content and genetic variability of banana accessions from the Musa germplasm collection held at Embrapa Cassava and Tropical Fruits, Brazil. Forty-two samples were analyzed, including 21 diploids, 19 triploids and two tetraploids. The carotenoid content was analyzed spectrophotometrically and genetic variability was estimated using 653 DArT markers. The average carotenoid content was 4.73 MUg.g (-1) , and ranged from 1.06 MUg.g (-1) for the triploid Nanica (Cavendish group) to 19.24 MUg.g ( 1) for the triploid Saney. The diploids Modok Gier and NBA-14 and the triploid Saney had a carotenoid content that was, respectively, 7-fold, 6-fold and 9-fold greater than that of cultivars from the Cavendish group (2.19 MUg.g (-1)). The mean similarity among the 42 accessions was 0.63 (range: 0.24 to 1.00). DArT analysis revealed extensive genetic variability in accessions from the Embrapa Musa germplasm bank. PMID- 21637653 TI - Genetic characterization of cassava (Manihot esculenta) landraces in Brazil assessed with simple sequence repeats. AB - Based on nine microsatellite loci, the aim of this study was to appraise the genetic diversity of 42 cassava (Manihot esculenta) landraces from selected regions in Brazil, and examine how this variety is distributed according to origin in several municipalities in the states of Minas Gerais, Sao Paulo, Mato Grosso do Sul, Amazonas and Mato Grosso. High diversity values were found among the five above-mentioned regions, with 3.3 alleles per locus on an average, a high percentage of polymorphic loci varying from 88.8% to 100%, an average of 0.265 for observed heterozygosity and 0.570 for gene diversity. Most genetic diversity was concentrated within the regions themselves (H(S) = 0.52). Cluster analysis and principal component based scatter plotting showed greater similarity among landraces from Sao Paulo, Mato Grosso do Sul and Amazonas, whereas those from Minas Gerais were clustered into a sub-group within this group. The plants from Mato Grosso, mostly collected in the municipality of General Carneiro, provided the highest differentiation. The migration of human populations is one among the possible reasons for this closer resemblance or greater disparity among plants from the various regions. PMID- 21637654 TI - The karyotype of Nothoscordum arenarium Herter (Gilliesioideae, Alliaceae): A populational and cytomolecular analysis. AB - The genus Nothoscordum Kunth comprises approximately 20 species native to South America. Karyologically, the genus is remarkable for its large chromosomes and Robertsonian translocations. Variation in chromosome number has been recorded in a few polyploid species and it is unknown among diploids. This study presents the chromosome number and morphology of 53 individuals of seven populations of N. arenarium Herter (2n = 10). In addition, karyotype analyses after C-banding, staining with CMA and DAPI, and in situ hybridization with 5S and 45S rDNA probes were performed in six individuals from one population. All individuals exhibited 2n = 10 (6M + 4A), except for one tetraploid (2n = 20, 12M + 8A) and one triploid (2n = 15, 9M + 6A) plant. C-banding revealed the presence of CMA(+) /DAPI (-) heterochromatin in the short arm and in the proximal region of the long arm of all acrocentric chromosomes. The 45S rDNA sites co-localized with the CMA (+) regions of the acrocentrics short arms, while the 5S rDNA probe only hybridized with the subterminal region of a pair of metacentric chromosomes. A change in the pattern of CMA bands and rDNA sites was observed in only one individual bearing a reciprocal translocation involving the long arm of a metacentric and the long arm of an acrocentric chromosome. These data suggest that, despite isolated cases of polyploidy and translocation, the karyotype of N. arenarium is very stable and the karyotypic instability described for other species may be associated with their polyploid condition. PMID- 21637655 TI - Characterization of six microsatellite loci in Myrica faya (Myricaceae) and cross amplification in the endangered endemic M. rivas-martinezii in Canary Islands, Spain. AB - Six novel polymorphic microsatellite markers were isolated from enriched libraries in Myrica faya Ait., recently renamed Morella faya, (fayatree, firetree, or firebush) in order to examine the genetic diversity in natural populations. Also, test cross-specific amplification and genetic diversity in Myrica rivas-martinezii, which is endemic on the Canary islands. Microsatellite loci were screened in 225 individuals of both species from different islands of the Canarian archipelago. All markers were successfully amplified from both Myrica species, with an average number of 6.5 and 9.3 alleles per locus in M. rivas-martinezii and M. faya, respectively. There was no evidence for linkage disequilibrium between loci, and the probability of null alleles ranged from 0.01 to 0.17. PMID- 21637656 TI - Comparative analysis of the genes UL1 through UL7 of the duck enteritis virus and other herpesviruses of the subfamily Alphaherpesvirinae. AB - The nucleotide sequences of eight open reading frames (ORFs) located at the 5' end of the unique long region of the duck enteritis virus (DEV) Clone-03 strain were determined. The genes identified were designated UL1, UL2, UL3, UL4, UL5, UL6 and UL7 homologues of the herpes simplex virus 1 (HSV-1). The DEV UL3.5 located between UL3 and UL4 had no homologue in the HSV-1. The arrangement and transcription orientation of the eight genes were collinear with their homologues in the HSV-1. Phylogenetic trees were constructed based on the alignments of the deduced amino acids of eight proteins with their homologues in 12 alpha herpesviruses. In the UL1, UL3, UL3.5, UL5 and UL7 proteins trees, the branches were more closely related to the genus Mardivirus. However, the UL2, UL4, and UL6 proteins phylogenetic trees indicated a large distance from Mardivirus, indicating that the DEV evolved differently from other viruses in the subfamily Alphaherpesvirinae and formed a single branch within this subfamily. PMID- 21637657 TI - Gel mobility shift scanning of pectin-inducible promoter from Penicillium griseoroseum reveals the involvement of a CCAAT element in the expression of a polygalacturonase gene. AB - Previous reports have described pgg2, a polygalacturonase-encoding gene of Penicillium griseoroseum, as an attractive model for transcriptional regulation studies, due to its high expression throughout several in vitro growth conditions, even in the presence of non-inducing sugars such as sucrose. A search for regulatory motifs in the 5' upstream regulatory sequence of pgg2 identified a putative CCAAT box that could justify this expression profile. This element, located 270 bp upstream of the translational start codon, was tested as binding target for regulatory proteins. Analysis of a 170 bp promoter fragment by electrophoretic mobility shift assay (EMSA) with nuclear extracts prepared from mycelia grown in pectin-containing culture medium revealed a high mobility complex that was subsequently confirmed by analyzing it with a double-stranded oligonucleotide spanning the CCAAT motif. A substitution in the core sequence for GTAGG partially abolished the formation of specific complexes, showing the involvement of the CCAAT box in the regulation of the polygalacturonase gene studied. PMID- 21637658 TI - Effect of organic tomato (Lycopersicon esculentum) extract on the genotoxicity of doxorubicin in the Drosophila wing spot test. AB - The consumption of organic tomatoes (ORTs) reduces the risk of harmful effects to humans and the environment caused by exposure to toxic agrochemicals. In this study, we used the somatic mutation and recombination test (SMART) of wing spots in Drosophila melanogaster to evaluate the genotoxicity of ORT and the effect of cotreatment with ORT on the genotoxicity of Doxorubicin((r)) (DXR, a cancer chemotherapeutic agent) that is mediated by free radical formation. Standard (ST) cross larvae were treated chronically with solutions containing 25%, 50% or 100% of an aqueous extract of ORT, in the absence and presence of DXR (0.125 mg/mL), and the number of mutant spots on the wings of emergent flies was counted. ORT alone was not genotoxic but enhanced the toxicity of DXR when administered concomitantly with DXR. The ORT-enhanced frequency of spots induced by DXR may have resulted from the interaction of ORT with the enzymatic systems that catalyze the metabolic detoxification of this drug. PMID- 21637659 TI - Profile of micronucleus frequencies and DNA damage in different species of fish in a eutrophic tropical lake. AB - Lake Paranoa is a tropical reservoir for the City of Brasilia, which became eutrophic due to inadequate sewage treatment associated with intensive population growth. At present, two wastewater treatment plants are capable of processing up to 95% of the domestic sewage, thereby successfully reducing eutrophization. We evaluated both genotoxic and cytotoxic parameters in several fish species (Geophagus brasiliensis, Cichla temensis, Hoplias malabaricus, Astyanax bimaculatus lacustres, Oreochromis niloticus, Cyprinus carpio and Steindachnerina insculpita) by using the micronucleus (MN) test, the comet assay and nuclear abnormality assessment in peripheral erythrocytes. The highest frequencies of MN were found in Cichla temensis and Hoplias malabaricus, which were statistically significant when compared to the other species. However, Steindachnerina insculpita (a detritivorous and lake-floor feeder species) showed the highest index of DNA damage in the comet assay, followed by C. temensis (piscivorous). Nuclear abnormalities, such as binucleated, blebbed, lobed and notched cells, were used as evidence of cytotoxicity. Oreochromis niloticus followed by Hoplias malaricus, ominivorous/detritivotous and piscivorous species, respectively, presented the highest frequency of nuclear abnormalities, especially notched cells, while the herbivorous Astyanax bimaculatus lacustres showed the lowest frequency compared to the other species studied. Thus, for biomonitoring aquatic genotoxins under field conditions, the food web should also be considered. PMID- 21637660 TI - Preferential induction of MLL(Mixed Lineage Leukemia) rearrangements in human lymphocyte cultures treated with etoposide. AB - Topoisomerase II inhibitors are effective chemotherapeutic agents in the treatment of cancer, in spite of being associated with the development of secondary leukemia. Our purpose was to determine the effects of etoposide on different genomic regions, aiming at discovering whether there are preferential sites which can be targeted by this drug in peripheral lymphocytes from healthy individuals. The in vitro treatment with low doses of etoposide (0.25, 0.5, and 1 MUg/mL, in 1 hour-pulse or continuous-48 h treatment) induced a significant increase in chromosomal aberrations, detected by conventional staining and FISH with specific probes for chromosomes 8 and 11, compared with untreated controls (p < 0.05). Additionally, the frequencies of alterations at 11q23, detected by MLL specific probes, were significantly higher (p < 0.005) in treated cells than in controls. In contrast, an analysis of rearrangements involving the IGH gene did not disclose differences between treatments. The present results demonstrated the potential of etoposide to interact with preferential chromosome sites in human lymphocytes, even at concentrations below the mean plasma levels measured in cancer patients. This greater susceptibility to etoposide-induced cleavage may explain the more frequent involvement of MLL in treatment-related leukemia. PMID- 21637661 TI - Cytogenotoxicity biomarkers in fat snook Centropomus parallelus from Cananeia and Sao Vicente estuaries, SP, Brazil. AB - The aquatic environment receives many contaminants that can induce damages at the molecular, biochemical, cellular and physiological levels. Centropomus parallelus, an important food resource for local populations, is a predator fish that feeds on small fishes and benthic invertebrates, thus being vulnerable to the bioconcentration and biomagnification processes. This study aimed to evaluate cytogenotoxic responses in erythrocytes from C. parallelus juveniles collected in the Cananeia and Sao Vicente estuaries, both in winter and in summer. After anesthesia, blood samples were collected by caudal puncture. Blood smears were prepared on glass slides and stained with May-Grunwald-Giemsa dye. Two thousand cells were analyzed per slide (1000x), and nuclear abnormalities (NA) and micronuclei (MN) were scored. The Sao Vicente sample showed MN and NA frequencies (%/1000 cells) of 0.325 and 3.575, in winter, and of 0.125 and 2.935 in summer respectively; the Cananeia sample showed frequencies of 0.0325 and 0.03, in winter, and of 0.065 and 0.355 in summer, respectively. The rates found in Sao Vicente were significantly higher than those found in Cananeia, evidencing that the levels of pollution in that estuary were high enough to induce genetic damages. PMID- 21637662 TI - Application of the comet assay in erythrocytes of Oreochromis niloticus (Pisces): A methodological comparison. AB - The present study applied the comet assay to erythrocytes of Oreochromis niloticus with the aim of improving protocols to detect DNA damage in these cells, by using two distinct pHs (pH = 12.1 and pH > 13) and evaluating whether there is a correspondence between silver and ethidium bromide staining. Comets were visually examined and, the frequency of cells with and without damage was obtained, as well as the distribution of classes and scores. By using the Kruskal Wallis test, our results revealed that pH 12.1 is more effective, although both pHs can be used. Our findings also suggest that silver staining can substitute ethidium bromide, an expensive and highly toxic stain that requires specific equipment for examination. PMID- 21637663 TI - Non-random pre-transcriptional evolution in HIV-1. A refutation of the foundational conditions for neutral evolution. AB - The complete base sequence of HIV-1 virus and GP120 ENV gene were analyzed to establish their distance to the expected neutral random sequence. An especial methodology was devised to achieve this aim. Analyses included: a) proportion of dinucleotides (signatures); b) homogeneity in the distribution of dinucleotides and bases (isochores) by dividing both segments in ten and three sub-segments, respectively; c) probability of runs of bases and No-bases according to the Bose Einstein distribution. The analyses showed a huge deviation from the random distribution expected from neutral evolution and neutral-neighbor influence of nucleotide sites. The most significant result is the tremendous lack of CG dinucleotides (p < 10(-50) ), a selective trait of eukaryote and not of single stranded RNA virus genomes. Results not only refute neutral evolution and neutral neighbor influence, but also strongly indicate that any base at any nucleotide site correlates with all the viral genome or sub-segments. These results suggest that evolution of HIV-1 is pan-selective rather than neutral or nearly neutral. PMID- 21637664 TI - Modeling body size evolution in Felidae under alternative phylogenetic hypotheses. AB - The use of phylogenetic comparative methods in ecological research has advanced during the last twenty years, mainly due to accurate phylogenetic reconstructions based on molecular data and computational and statistical advances. We used phylogenetic correlograms and phylogenetic eigenvector regression (PVR) to model body size evolution in 35 worldwide Felidae (Mammalia, Carnivora) species using two alternative phylogenies and published body size data. The purpose was not to contrast the phylogenetic hypotheses but to evaluate how analyses of body size evolution patterns can be affected by the phylogeny used for comparative analyses (CA). Both phylogenies produced a strong phylogenetic pattern, with closely related species having similar body sizes and the similarity decreasing with increasing distances in time. The PVR explained 65% to 67% of body size variation and all Moran's I values for the PVR residuals were non-significant, indicating that both these models explained phylogenetic structures in trait variation. Even though our results did not suggest that any phylogeny can be used for CA with the same power, or that "good" phylogenies are unnecessary for the correct interpretation of the evolutionary dynamics of ecological, biogeographical, physiological or behavioral patterns, it does suggest that developments in CA can, and indeed should, proceed without waiting for perfect and fully resolved phylogenies. PMID- 21637665 TI - Alcohol dehydrogenase activities and ethanol tolerance in Anastrepha (Diptera, Tephritidae) fruit-fly species and their hybrids. AB - The ADH (alcohol dehydrogenase) system is one of the earliest known models of molecular evolution, and is still the most studied in Drosophila. Herein, we studied this model in the genus Anastrepha (Diptera, Tephritidae). Due to the remarkable advantages it presents, it is possible to cross species with different Adh genotypes and with different phenotype traits related to ethanol tolerance. The two species studied here each have a different number of Adh gene copies, whereby crosses generate polymorphisms in gene number and in composition of the genetic background. We measured certain traits related to ethanol metabolism and tolerance. ADH specific enzyme activity presented gene by environment interactions, and the larval protein content showed an additive pattern of inheritance, whilst ADH enzyme activity per larva presented a complex behavior that may be explained by epistatic effects. Regression models suggest that there are heritable factors acting on ethanol tolerance, which may be related to enzymatic activity of the ADHs and to larval mass, although a pronounced environmental effect on ethanol tolerance was also observed. By using these data, we speculated on the mechanisms of ethanol tolerance and its inheritance as well as of associated traits. PMID- 21637666 TI - On the reliability of a simple method for scoring phenotypes to estimate heritability: A case study with pupal color in Heliconius erato phyllis, Fabricius 1775 (Lepidoptera, Nymphalidae). AB - In this paper, two methods for assessing the degree of melanization of pupal exuviae from the butterfly Heliconius erato phyllis, Fabricius 1775 (Lepidoptera, Nymphalidae, Heliconiini) are compared. In the first method, which was qualitative, the exuviae were classified by scoring the degree of melanization, whereas in the second method, which was quantitative, the exuviae were classified by optical density followed by analysis with appropriate software. The heritability (h(2)) of the degree of melanization was estimated by regression and analysis of variance. The estimates of h (2) were similar with both methods, indicating that the qualitative method could be particularly suitable for field work. The low estimates obtained for heritability may have resulted from the small sample size (n = 7-18 broods, including the parents) or from the allocation priority hypothesis in which pupal color would be a lower priority trait compared to morphological traits and adequate larval development. PMID- 21637667 TI - High levels of genetic variability and differentiation in hilsa shad, Tenualosa ilisha (Clupeidae, Clupeiformes) populations revealed by PCR-RFLP analysis of the mitochondrial DNA D-loop region. AB - The hilsa shad, Tenualosa ilisha (Clupeidae, Clupeiformes) is an important anadromous clupeid species from the Western division of the Indo-Pacific region. It constitutes the largest single fishable species in Bangladesh. Information on genetic variability and population structure is very important for both management and conservation purposes. Past reports on the population structure of T. ilisha involving morphometric, allozyme and RAPD analyses are contradictory. We examined genetic variability and divergence in two riverine (the Jamuna and the Meghna), two estuarine (Kuakata and Sundarbans) and one marine (Cox's Bazar) populations of T. ilisha by applying PCR-RFLP analysis of the mtDNA D-loop region. The amplified PCR products were restricted with four restriction enzymes namely, XbaI, EcoRI, EcoRV, and HaeIII. High levels of haplotype and gene diversity within and significant differentiations among, populations of T. ilisha were observed in this study. Significant F(ST) values indicated differentiation among the river, estuary and marine populations. The UPGMA dendrogram based on genetic distance resulted in two major clusters, although, these were subsequently divided into three, corresponding to the riverine, estuarine and marine populations. The study underlines the usefulness of RFLP of mtDNA D-loop region as molecular markers, and detected at least two differentiated populations of T. ilisha in Bangladesh waters. PMID- 21637668 TI - Genetic variation of wild and hatchery populations of the catla Indian major carp (Catla catla Hamilton 1822: Cypriniformes, Cyprinidae) revealed by RAPD markers. AB - Genetic variation is a key component for improving a stock through selective breeding programs. Randomly amplified polymorphic DNA (RAPD) markers were used to assess genetic variation in three wild population of the catla carp (Catla catla Hamilton 1822) in the Halda, Jamuna and Padma rivers and one hatchery population in Bangladesh. Five decamer random primers were used to amplify RAPD markers from 30 fish from each population. Thirty of the 55 scorable bands were polymorphic, indicating some degree of genetic variation in all the populations. The proportion of polymorphic loci and gene diversity values reflected a relatively higher level of genetic variation in the Halda population. Sixteen of the 30 polymorphic loci showed a significant (p < 0.05, p < 0.01, p < 0.001) departure from homogeneity and the F(ST) values in the different populations indicated some degree of genetic differentiation in the population pairs. Estimated genetic distances between populations were directly correlated with geographical distances. The unweighted pair group method with averages (UPGMA) dendrogram showed two clusters, the Halda population forming one cluster and the other populations the second cluster. Genetic variation of C. catla is a useful trait for developing a good management strategy for maintaining genetic quality of the species. PMID- 21637669 TI - A review of techniques for spatial modeling in geographical, conservation and landscape genetics. AB - Most evolutionary processes occur in a spatial context and several spatial analysis techniques have been employed in an exploratory context. However, the existence of autocorrelation can also perturb significance tests when data is analyzed using standard correlation and regression techniques on modeling genetic data as a function of explanatory variables. In this case, more complex models incorporating the effects of autocorrelation must be used. Here we review those models and compared their relative performances in a simple simulation, in which spatial patterns in allele frequencies were generated by a balance between random variation within populations and spatially-structured gene flow. Notwithstanding the somewhat idiosyncratic behavior of the techniques evaluated, it is clear that spatial autocorrelation affects Type I errors and that standard linear regression does not provide minimum variance estimators. Due to its flexibility, we stress that principal coordinate of neighbor matrices (PCNM) and related eigenvector mapping techniques seem to be the best approaches to spatial regression. In general, we hope that our review of commonly used spatial regression techniques in biology and ecology may aid population geneticists towards providing better explanations for population structures dealing with more complex regression problems throughout geographic space. PMID- 21637670 TI - HLA-DRB1 alleles in four Amerindian populations from Argentina and Paraguay. AB - The major histocompatibility complex (MHC) is one of the biological systems of major polymorphisms. The study of HLA class II variability has allowed the identification of several alleles that are characteristic to Amerindian populations, and it is an excellent tool to define the relations and biological affinities among them. In this work, we analyzed the allelic distribution of the HLA-DRB1 class II locus in four Amerindian populations: Mapuche (n = 34) and Tehuelche (n = 23) from the Patagonian region of Argentina, and Wichi SV (n = 24) and Lengua (n = 17) from the Argentinean and Paraguayan Chaco regions, respectively. In all of these groups, relatively high frequencies of Amerindian HLA-DRB1 alleles were observed (DRB1*0403, DRB1*0407, DRB1*0411, DRB1*0417, DRB1*0802, DRB1*0901, DRB1*1402, DRB1*1406 and DRB1*1602). However, we also detected the presence of non-Amerindian variants in Mapuche (35%) and Tehuelche (22%). We compared our data with those obtained in six indigenous groups of the Argentinean Chaco region and in a sample from Buenos Aires City. The genetic distance dendrogram showed a clear-cut division between the Patagonian and Chaco populations, which formed two different clusters. In spite of their linguistic differences, it can be inferred that the biological affinities observed are in concordance with the geographic distributions and interethnic relations established among the groups studied. PMID- 21637671 TI - Linkage disequilibrium and diversity for three genomic regions in Azoreans and mainland Portuguese. AB - Studies on linkage disequilibrium (LD) across the genome and populations have been used in recent years with the main objective of improving gene mapping of complex traits. Here, we characterize the patterns of genetic diversity of HLA loci and evaluate LD (D') extent in three genomic regions: Xq13.3, NRY and HLA. In addition, we examine the distribution of DXS1225-DXS8082 haplotype diversity in Azoreans and mainland Portuguese. Allele distribution has demonstrated that the Sao Miguel population is genetically very diverse; haplotype analysis revealed 100% discriminatory power for X- and Y-markers and 94.3% for HLA markers. Standardized multiallelic D' in these three genomic regions shows values lower than 0.33, thereby suggesting there is no extensive LD in the Sao Miguel population. Data regarding the distribution of DXS1225-DXS8082 haplotypes indicate that there are no significant differences among all the populations studied, (Azorean geographical groups, the Azores archipelago and mainland Portugal). Moreover, in these as well as in other European populations, the most frequent DXS1225-DXS8082 haplotype is 210-219. Even though Sao Miguel islanders and Azoreans do not constitute isolated populations and show LD for only very short physical distances, certain characteristics, such as the absence of genetic structure, the same environment and the possibility of constructing extensive pedigrees through church and civil records, offer an opportunity for dissecting the genetic background of complex diseases in these populations. PMID- 21637672 TI - Effects of APOE, APOB and LDLR variants on serum lipids and lack of association with xanthelasma in individuals from Southeastern Brazil. AB - Xanthelasma might be a clinical manifestation of dyslipidemia, a recognized risk factor for coronary artery disease. We investigated the association of apolipoprotein E (APOE HhaI), apolipoprotein B (APOB XbaI and Ins/Del) and LDL receptor (LDLR AvaII and HincII) gene polymorphisms with lipid profiles in 100 Brazilians with xanthelasma and 100 controls. Allele frequencies were similar in both groups. APOE, APOB and LDLR genotypes were not correlated with differences in the serum lipid profile. In individuals with xanthelasma, the APOB D allele was associated with less chance of having increased LDL-cholesterol (O.R. = 0.16, CI95% = 0.03-0.94, p = 0.042). In the control group, the APOB X+ allele was associated with less chance of having both increased total cholesterol (O.R. = 0.16, CI95% = 0.03-0.78, p = 0.023) and increased LDL-cholesterol (O.R. = 0.10, CI95% = 0.02-0.60, p = 0.012). Moreover, there was a significantly higher frequency of control individuals (68%) with elevated serum triglyceride levels, compared to patients (48%, p = 0.008). On the other hand, triglyceride levels in controls also seemed to be influenced by all other gene polymorphisms studied, an effect that might be enhanced by environmental factors. PMID- 21637673 TI - MLL leukemia-associated rearrangements in peripheral blood lymphocytes from healthy individuals. AB - Chromosomal translocations are characteristic of hematopoietic neoplasias and can lead to unregulated oncogene expression or the fusion of genes to yield novel functions. In recent years, different lymphoma/leukemia-associated rearrangements have been detected in healthy individuals. In this study, we used inverse PCR to screen peripheral lymphocytes from 100 healthy individuals for the presence of MLL (Mixed Lineage Leukemia) translocations. Forty-nine percent of the probands showed MLL rearrangements. Sequence analysis showed that these rearrangements were specific for MLL translocations that corresponded to t(4;11)(q21;q23) (66%) and t(9;11) (20%). However, RT-PCR failed to detect any expression of t(4;11)(q21;q23) in our population. We suggest that 11q23 rearrangements in peripheral lymphocytes from normal individuals may result from exposure to endogenous or exogenous DNA-damaging agents. In practical terms, the high susceptibility of the MLL gene to chemically-induced damage suggests that monitoring the aberrations associated with this gene in peripheral lymphocytes may be a sensitive assay for assessing genomic instability in individuals exposed to genotoxic stress. PMID- 21637674 TI - Peripheral position of CCND1 and HER-2/neu oncogenes within chromosome territories in esophageal and gastric cancers non-related to amplification and overexpression. AB - Interphase chromosomes have been shown to occupy discrete regions of the nucleus denominated chromosome territories (CTs), their active genes being preferentially positioned on the surfaces of these CTs, where they are accessible to transcriptional machinery. By means of FISH (Fluorescence in situ Hybridization), we analyzed the CCND1 and HER-2/neu gene positions within the CTs and their relationship with gene amplification and protein over-expression in esophageal and gastric cancers. The CCND1 and HER-2/Neu genes were more often positioned at the periphery (mean frequency of 60%-83%) of the CTs in tumor tissues of the esophagus and stomach. Moreover, this positioning revealed no association with either gene amplification or the protein over-expression status of these genes, although, in esophageal carcinoma, Kappa statistics showed a moderate agreement between amplification of the CCND1 gene (Kappa = 0.400) and its location within the CT, as well as with over-expression of the corresponding protein (Kappa = 0.444). Thus, our results suggest that gene positioning in interphase chromosomes does not follow a definitive pattern neither does it depend only on gene transcriptional activity. Apparently, this positioning could be both gene- and tissue-specific, and depends on other factors acting together, such as dense gene, chromosome size, chromatin structure, and the level and stability of its expression. PMID- 21637675 TI - A novel point mutation in a class IV glucose-6-phosphate dehydrogenase variant (G6PD Sao Paulo) and polymorphic G6PD variants in Sao Paulo State, Brazil. AB - In this study, we used red cell glucose-6-phosphate dehydrogenase (G6PD) activity to screen for G6PD-deficient individuals in 373 unrelated asymptomatic adult men who were working with insecticides (organophosphorus and carbamate) in dengue prevention programs in 27 cities in Sao Paulo State, Brazil. Twenty-one unrelated male children suspected of having erythroenzymopathy who were attended at hospitals in Sao Paulo city were also studied. Fifteen of the 373 adults and 12 of the 21 children were G6PD deficient. G6PD gene mutations were investigated in these G6PD-deficient individuals by using PCR-RFLP, PCR-SSCP analysis and DNA sequencing. Twelve G6PD A-202A/376G and two G6PD Seattle844C, as well as a new variant identified as G6PD Sao Paulo, were detected among adults, and 11 G6PD A 202A/376G and one G6PD Seattle844C were found among children. The novel mutation c.660C > G caused the replacement of isoleucine by methionine (I220M) in a region near the dimer interface of the molecule. The conservative nature of this mutation (substitution of a nonpolar aliphatic amino acid for another one) could explain why there was no corresponding change in the loss of G6PD activity (64.5% of normal activity in both cases). PMID- 21637676 TI - XRCC1 gene polymorphisms in a population sample and in women with a family history of breast cancer from Rio de Janeiro (Brazil). AB - The X-ray repair cross-complementing Group1 (XRCC1) gene has been defined as essential in the base excision repair (BER) and single-strand break repair processes. This gene is highly polymorphic, and the most extensively studied genetic changes are in exon 6 (Arg194Trp) and in exon 10 (Arg399Gln). These changes, in conserved protein sites, may alter the base excision repair capacity, increasing the susceptibility to adverse health conditions, including cancer. In the present study, we estimated the frequencies of the XRCC1 gene polymorphisms Arg194Trp and Arg399Gln in healthy individuals and also in women at risk of breast cancer due to family history from Rio de Janeiro. The common genotypes in both positions (194 and 399) were the most frequent in this Brazilian sample. Although the 194Trp variant was overrepresented in women reporting familial cases of breast cancer, no statistically significant differences concerning genotype distribution or intragenic interactions were found between this group and the controls. Thus, in the population analyzed by us, variants Arg194Trp and Arg399Gln did not appear to have any impact on breast cancer susceptibility. PMID- 21637677 TI - Myeloperoxidase gene-463G > A polymorphism and premature coronary artery disease. AB - We investigated the association between myeloperoxidase gene -463G > A polymorphism and premature coronary artery disease (CAD) in two Chinese population samples: 229 patients and 230 controls. Genotypes were determined by ligase detection reaction-polymerase chain reaction sequencing and the grouping technique. We found lower frequencies of both the A/A genotype and the A allele in patients (p < 0.05). Multivariate logistic regression showed that the risk of premature CAD in subjects carrying the AA genotype was reduced by 83% in relation to individuals carrying the G/G genotype (OR = 0.172, 95% CI: 0.057-0.526, p = 0.002). Our results indicate that -463G > A polymorphism of the myeloperoxidase gene is associated with premature CAD in Chinese individuals, suggesting that the AA genotype is a protective factor against premature CAD. PMID- 21637678 TI - Increased risk of venous thrombosis by AB alleles of the ABO blood group and Factor V Leiden in a Brazilian population. AB - Most cases of a predisposition to venous thrombosis are caused by resistance to activated protein C, associated in 95% of cases with the Factor V Leiden allele (FVL or R506Q). Several recent studies report a further increased risk of thrombosis by an association between the AB alleles of the ABO blood group and Factor V Leiden. The present study investigated this association with deep vein thrombosis (DVT) in individuals treated at the Hemocentro de Pernambuco in northeastern Brazil. A case-control comparison showed a significant risk of thrombosis in the presence of Factor V Leiden (OR = 10.1), which was approximately doubled when the AB alleles of the ABO blood group were present as well (OR = 22.3). These results confirm that the increased risk of deep vein thrombosis in the combined presence of AB alleles and Factor V Leiden is also applicable to the Brazilian population suggesting that ABO blood group typing should be routinely added to FVL in studies involving thrombosis. PMID- 21637679 TI - Karyotype and genome size of Iberochondrostoma almacai (Teleostei, Cyprinidae) and comparison with the sister-species I.lusitanicum. AB - This study aimed to define the karyotype of the recently described Iberian endemic Iberochondrostoma almacai, to revisit the previously documented chromosome polymorphisms of its sister species I.lusitanicum using C-, Ag-/CMA(3) and RE-banding, and to compare the two species genome sizes. A 2n = 50 karyotype (with the exception of a triploid I.lusitanicum specimen) and a corresponding haploid chromosome formula of 7M:15SM:3A (FN = 94) were found. Multiple NORs were observed in both species (in two submetacentric chromosome pairs, one of them clearly homologous) and a higher intra and interpopulational variability was evidenced in I.lusitanicum. Flow cytometry measurements of nuclear DNA content showed some significant differences in genome size both between and within species: the genome of I. almacai was smaller than that of I.lusitanicum (mean values 2.61 and 2.93 pg, respectively), which presented a clear interpopulational variability (mean values ranging from 2.72 to 3.00 pg). These data allowed the distinction of both taxa and confirmed the existence of two well differentiated groups within I. lusitanicum: one that includes the populations from the right bank of the Tejo and Samarra drainages, and another that reunites the southern populations. The peculiar differences between the two species, presently listed as "Critically Endangered", reinforced the importance of this study for future conservation plans. PMID- 21637680 TI - Comparative cytogenetics of three species of Dichotomius (Coleoptera, Scarabaeidae). AB - Meiotic and mitotic chromosomes of Dichotomius nisus, D. semisquamosus and D. sericeus were analyzed after conventional staining, C-banding and silver nitrate staining. In addition, Dichotomius nisus and D. semisquamosus chromosomes were also analyzed after fluorescent in situ hybridization (FISH) with an rDNA probe. The species analyzed had an asymmetrical karyotype with 2n = 18 and meta submetacentric chromosomes. The sex determination mechanism was of the Xy(p) type in D. nisus and D. semisquamosus and of the Xy (r) type in D. sericeus. C-banding revealed the presence of pericentromeric blocks of constitutive heterochromatin (CH) in all the chromosomes of the three species. After silver staining, the nucleolar organizer regions (NORs) were located in autosomes of D. semisquamosus and D. sericeus and in the sexual bivalent of D. nisus. FISH with an rDNA probe confirmed NORs location in D. semisquamosus and in D. nisus. Our results suggest that chromosome inversions and fusions occurred during the evolution of the group. PMID- 21637681 TI - Genotype by environment interaction for 450-day weight of Nelore cattle analyzed by reaction norm models. AB - Genotype by environment interactions (GEI) have attracted increasing attention in tropical breeding programs because of the variety of production systems involved. In this work, we assessed GEI in 450-day adjusted weight (W450) Nelore cattle from 366 Brazilian herds by comparing traditional univariate single-environment model analysis (UM) and random regression first order reaction norm models for six environmental variables: standard deviations of herd-year (RRMw) and herd year-season-management (RRMw-m) groups for mean W450, standard deviations of herd year (RRMg) and herd-year-season-management (RRMg-m) groups adjusted for 365-450 days weight gain (G450) averages, and two iterative algorithms using herd-year season-management group solution estimates from a first RRMw-m and RRMg-m analysis (RRMITw-m and RRMITg-m, respectively). The RRM results showed similar tendencies in the variance components and heritability estimates along environmental gradient. Some of the variation among RRM estimates may have been related to the precision of the predictor and to correlations between environmental variables and the likely components of the weight trait. GEI, which was assessed by estimating the genetic correlation surfaces, had values < 0.5 between extreme environments in all models. Regression analyses showed that the correlation between the expected progeny differences for UM and the corresponding differences estimated by RRM was higher in intermediate and favorable environments than in unfavorable environments (p < 0.0001). PMID- 21637682 TI - Genetic parameters for faecal egg count, packed-cell volume and body-weight in Santa Ines lambs. AB - Worm infection is one of the main factors responsible for economic losses in sheep breeding in Brazil. Random regression analysis was used to estimate genetic parameters for the factors faecal egg-count (FEC), packed-cell volume (PCV) and body weight (BW) in Santa Ines lambs. Data from 119 female, offspring of nine rams, were collected between December, 2005 and December, 2006, from the experimental flock of Embrapa Tabuleiros Costeiros, the Brazilian Agricultural Research Corporation located in Frei Paulo, SE, Brazil. After weaning, females were drenched until the faecal egg count had dropped to zero. Two natural challenges were undertaken. FEC heritability was extremely variable, this increasing from 0.04 to 0.27 in the first challenge and from 0.01 to 0.52 during the second. PCV heritability peaks were 0.31 and 0.12 in the first and second challenges, respectively. In the second challenge, BW heritability was close to 0.90. The genetic correlations among these traits did not differ from zero. There is the possibility of increasing parasite resistance in Santa Ines by selecting those animals with lower FEC. Selection to increase resistance will not adversely affect lamb-growth, although lambs with a slow growth-rate may be more susceptible to infection. PMID- 21637683 TI - Characterization analysis and polymorphism detection of the porcine Myd88 gene. AB - The myeloid differentiation primary response protein 88 (Myd88) is an essential adaptor protein, which mediates in all Toll-like receptor (TLR) members signal transduction, except for TLR3. In this study, the 4464 bp genomic sequence of porcine Myd88 was first isolated, whereupon tissue distribution, chromosome mapping and single nucleotide polymorphism (SNP) were analyzed. Our results revealed that porcine Myd88 gene, which was located at chromosome 13 linked with marker S0288 (distance = 40 cR; LOD = 8.66), was widely expressed in all the examined tissues. There were 16 potential SNPs in the isolated genome fragment. SNP 797T/C in the first intron was studied, with no significant association being found between the genotype and immune traits in pigs (p > 0.05). The porcine Myd88 protein contained both the death domain (DD) and the Toll/IL-1 receptor domain (TIR). Leu residues, essential for its structure, were the most abundant encountered in the DD. The TIR contained two conserved motifs which may play important roles in the Myd88 function. PMID- 21637684 TI - Karyology of the Atlantic forest rodent Juliomys (Cricetidae): A new karyotype from southern Brazil. AB - Juliomys is a small rodent from the family Cricetidae which inhabits the Atlantic forest and forests from Argentina to eastern Brazil. The three species recognized so far have different karyotypes. In this paper, we describe a new karyotype with 2n = 32, FN = 48 found in Juliomys specimens from a high-altitude area in the Atlantic forest of southern Brazil. The karyotype was analyzed after G- and C banding and silver staining of the nucleolus organizer regions (Ag-NOR) and its G banding patterns were compared with those of the newly described species Juliomys ossitenuis (2n = 20, FN = 36). The 2n = 32 karyomorph presented peculiar features and was very different from those of the other species of the genus: J. pictipes (2n = 36, FN = 34), J. rimofrons (2n = 20, FN = 34) and J. ossitenuis (2n = 20, FN = 36). Differences were mostly due to centric and tandem fusions, pericentric inversion and loss of heterochromatin. The karyotype represents a powerful tool to differentiate Juliomys species and our data suggest that the karyotype described herein belongs to a new species. PMID- 21637685 TI - Implementation of a parentage control system in Portuguese beef-cattle with a panel of microsatellite markers. AB - A study was conducted to assess the feasibility of applying a panel of 10 microsatellite markers in parentage control of beef cattle in Portugal. In the first stage, DNA samples were collected from 475 randomly selected animals of the Charolais, Limousin and Preta breeds. Across breeds and genetic markers, means for average number of alleles, effective number of alleles, expected heterozygosity and polymorphic information content, were 8.20, 4.43, 0.733 and 0.70, respectively. Enlightenment from the various markers differed among breeds, but the set of 10 markers resulted in a combined probability above 0.9995 in the ability to exclude a random putative parent. The marker-set thus developed was later used for parentage control in a group of 140 calves from several breeds, where there was the suspicion of possible faulty parentage recording. Overall, 76.4% of the calves in this group were compatible with the recorded parents, with most incompatibilities due to misidentification of the dam. Efforts must be made to improve the quality of pedigree information, with particular emphasis on information recorded at the calf's birth. PMID- 21637686 TI - Use of AFLP and RAPD molecular genetic markers and cytogenetic analysis to explore relationships among taxa of the Patagonian Bromus setifolius complex. AB - Bromus setifolius var. pictus (Hook) Skottsb., B. setifolius var. setifolius Presl. and B.setifolius var. brevifolius Ness are three native Patagonian taxa in the section Pnigma Dumort of the genus Bromus L. AFLP and RAPD analysis, in conjunction with genetic distance measurements and statistical techniques, revealed variation within this group and indicated that B. setifolius var. brevifolius was closely related to B. setifolius var. pictus, with both taxa being more distantly related to B. setifolius var. setifolius. Cytogenetic analysis confirmed the chromosomal number of B. setifolius var. pictus (2n = 70) and B. setifolius var. setifolius (2n = 28) and showed for the first time that B. setifolius var. brevifolius had 2n = 70. The combination of molecular genetic and cytogenetic evidence supported a species status for two of the three taxa and suggested hypotheses for the evolutionary origin of these complex taxa. Species status was also indicated for B. setifolius var. setifolius. Based on these findings, we suggest that B. setifolius var. pictus be referred to as B. pictus Hook var. pictus, and B. setifolius var brevifolius as B. pictus Hook var brevifolius. The correlation between AFLP diversity and variation in ecological parameters suggested that this marker system could be used to assess breeding progress and to monitor the domestication of Patagonian Bromus species for agronomic use. PMID- 21637687 TI - Karyotype differentiation of four Cestrum species (Solanaceae) revealed by fluorescent chromosome banding and FISH. AB - The karyotypes of four South American species of Cestrum (C. capsulare,C. corymbosum,C. laevigatum and C. megalophylum) were studied using conventional staining, C-CMA/DAPI chromosome banding and FISH with 45S and 5S rDNA probes. The karyotypes showed a chromosome number of 2n = 2x = 16, with metacentric chromosomes, except for the eighth submeta- to acrocentric pair. Several types of heterochromatin were detected, which varied in size, number, distribution and base composition. The C-CMA(+) bands and 45S rDNA were located predominantly in terminal regions. The C-CMA (+) /DAPI (+) bands appeared in interstitial and terminal regions, and the C-DAPI (+) bands were found in all chromosome regions. The 5S rDNA sites were observed on the long arm of pair 8 in all species except C. capsulare, where they were found in the paracentromeric region of the long arm of pair 4. The differences in band patterns among the species studied here, along with data from other nine species reported in the literature, suggest that the bands are dispersed in an equilocal and non-equilocal manner and that structural rearrangements can be responsible for internal karyotype diversification. However, it is important to point out that the structural changes involving repetitive segments did not culminate in substantial changes in the general karyotype structure concerning chromosome size and morphology. PMID- 21637688 TI - Genetic parameters and selection strategies for soybean genotypes resistant to the stink bug-complex. AB - Soybean genotypes resistant to stink bugs are derived from complex breeding processes obtained through indirect selection. The aim of the present work was to estimate genetic parameters for guiding selection strategies towards resistant genotypes, based on those traits associated with responses to pod-attacking stink bugs, such as the grain filling period (GFP), leaf retention (LR), percentage index of pod damage (PIPD) and percentage of spotted seeds (PSS). We assessed the parental lines IAC-100 (resistant) and FT-Estrela (susceptible), the progenies F(2) and F (4) , 30 progenies F (2:3) , 30 progenies BC (1) F (2:3) and 30 progenies BC (2) F (2:3) , besides the cultivars BRS Celeste and MGBR-46 (Conquista). Three field experiments, using randomized complete block design with three replications, were installed in Goiania-GO, in the 2002/03 season. Each experiment consisted of 36 treatments (6 common and 30 regular). Heritability estimates were: 74.6 and 36.1 (GFP); 51.9 and 19.9 (LR); 49.6 and 49.6 (PIPD) and 55.8 and 20.3 (PSS), in both the broad and narrow senses, respectively. Based on these results, we concluded that the best strategy for obtaining stink bug resistant genotypes consists of selecting the PIPD trait in early generations (F (3) or F (4) ), followed by selection for the GFP, LR and PSS traits in generations with higher endogamy levels. PMID- 21637689 TI - Cross-amplification and characterization of microsatellite loci for the Neotropical orchid genus Epidendrum. AB - In this study we tested the cross-amplification of 33 microsatellite loci previously developed for two closely related Neotropical orchid genera (Epidendrum and Laelia). A set of ten loci were polymorphic across five examined species (20 individuals each) with 2 to 15 alleles per locus. The mean expected and observed heterozygosity (average across species) ranged from 0.34 to 0.82 and from 0.27 to 0.85, respectively. In addition we tested all loci in 35 species representative of the genus Epidendrum. Of these, 26 loci showed successful amplification. Cross-application of these loci represent a potential source of co dominant markers for evolutionary, ecological and conservation studies in this important orchid genus. PMID- 21637690 TI - Xylella fastidiosa gene expression analysis by DNA microarrays. AB - Xylella fastidiosa genome sequencing has generated valuable data by identifying genes acting either on metabolic pathways or in associated pathogenicity and virulence. Based on available information on these genes, new strategies for studying their expression patterns, such as microarray technology, were employed. A total of 2,600 primer pairs were synthesized and then used to generate fragments using the PCR technique. The arrays were hybridized against cDNAs labeled during reverse transcription reactions and which were obtained from bacteria grown under two different conditions (liquid XDM(2) and liquid BCYE). All data were statistically analyzed to verify which genes were differentially expressed. In addition to exploring conditions for X. fastidiosa genome-wide transcriptome analysis, the present work observed the differential expression of several classes of genes (energy, protein, amino acid and nucleotide metabolism, transport, degradation of substances, toxins and hypothetical proteins, among others). The understanding of expressed genes in these two different media will be useful in comprehending the metabolic characteristics of X. fastidiosa, and in evaluating how important certain genes are for the functioning and survival of these bacteria in plants. PMID- 21637691 TI - Neurospora crassa mat A-2 and mat A-3 proteins weakly interact in the yeast two hybrid system and affect yeast growth. AB - Mating-type genes control the entry into the sexual cycle, mating identity and sexual development in fungi. The mat A-2 and mat A-3 genes, present in the mat A idiomorph of the filamentous fungus Neurospora crassa, are required for post fertilization functions but are not essential for mating identity. Their putative roles as transcription factors are based on the similarity of mat A-2 with the Podospora anserina SMR1 gene and an HMG motif present in the mat A-3 gene. In this work the yeast two-hybrid system was used to identify transcriptional activity and protein-protein interaction of N. crassamat A-2 and mat A-3 genes. We observed that the mat A-3 protein alone is capable of weakly activating transcription of yeast reporter genes; it also binds with low specificity to the GAL1 promoter sequence, possibly due to its HMG domain. Our results also indicate that mat A-3 is capable to form homodimers, and interact with mat A-2. Interference on yeast growth was observed on some transformants suggesting a toxic action of the mat A-2 protein. Our data on pattern of interactions of mat proteins contributes towards understanding the control of vegetative and sexual cycles in filamentous fungi. PMID- 21637692 TI - The glyceraldehyde-3-phosphate dehydrogenase gene of Moniliophthoraperniciosa, the causal agent of witches' broom disease of Theobroma cacao. AB - This report describes the cloning, sequence and expression analysis of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene of Moniliophthora perniciosa, the most important pathogen of cocoa in Brazil. Southern blot analysis revealed the presence of a single copy of the GAPDH gene in the M. perniciosa genome (MpGAPDH). The complete MpGAPDH coding sequence contained 1,461 bp with eight introns that were conserved in the GAPDH genes of other basidiomycete species. The cis-elements in the promoter region of the MpGAPDH gene were similar to those of other basidiomycetes. Likewise, the MpGAPDH gene encoded a putative 339 amino acid protein that shared significant sequence similarity with other GAPDH proteins in fungi, plants, and metazoans. Phylogenetic analyses clustered the MPGAPDH protein with other homobasidiomycete fungi of the family Tricholomataceae. Expression analysis of the MpGAPDH gene by real-time PCR showed that this gene was more expressed (~1.3X) in the saprotrophic stage of this hemibiotrophic plant pathogen than in the biotrophic stage when grown in cacao extracts. PMID- 21637693 TI - Estimates of DNA strand breakage in bottlenose dolphin (Tursiops truncatus) leukocytes measured with the Comet and DNA diffusion assays. AB - The analysis of DNA damage by mean of Comet or single cell gel electrophoresis (SCGE) assay has been commonly used to assess genotoxic impact in aquatic animals being able to detect exposure to low concentrations of contaminants in a wide range of species. The aims of this work were 1) to evaluate the usefulness of the Comet to detect DNA strand breakage in dolphin leukocytes, 2) to use the DNA diffusion assay to determine the amount of DNA strand breakage associated with apoptosis or necrosis, and 3) to determine the proportion of DNA strand breakage that was unrelated to apoptosis and necrosis. Significant intra-individual variation was observed in all of the estimates of DNA damage. DNA strand breakage was overestimated because a considerable amount (~29%) of the DNA damage was derived from apoptosis and necrosis. The remaining DNA damage in dolphin leukocytes was caused by factors unrelated to apoptosis and necrosis. These results indicate that the DNA diffusion assay is a complementary tool that can be used together with the Comet assay to assess DNA damage in bottlenose dolphins. PMID- 21637694 TI - Genotoxicity assessment of a pharmaceutical effluent using four bioassays. AB - Pharmaceutical industries are among the major contributors to industrial waste. Their effluents when wrongly handled and disposed of endanger both human and environmental health. In this study, we investigated the potential genotoxicity of a pharmaceutical effluent, by using the Allium cepa, mouse- sperm morphology, bone marrow chromosome aberration (CA) and micronucleus (MN) assays. Some of the physico-chemical properties of the effluent were also determined. The A. cepa and the animal assays were respectively carried out at concentrations of 0.5, 1, 2.5, 5 and 10%; and 1, 5, 10, 25 and 50% of the effluent. There was a statistically different (p < 0.05), concentration-dependent inhibition of onion root growth and mitotic index, and induction of chromosomal aberrations in the onion and mouse CA test. Assessment of sperm shape showed that the fraction of the sperm that was abnormal in shape was significantly (p < 0.05) greater than the negative control value. MN analysis showed a dose-dependent induction of micronucleated polychromatic erythrocytes across the treatment groups. These observations were provoked by the toxic and genotoxic constituents present in test samples. The tested pharmaceutical effluent is a potentially genotoxic agent and germ cell mutagen, and may induce adverse health effects in exposed individuals. PMID- 21637695 TI - Modulatory effects of Tabebuia impetiginosa (Lamiales, Bignoniaceae) on doxorubicin-induced somatic mutation and recombination in Drosophila melanogaster. AB - The wing Somatic Mutation and Recombination Test (SMART) in D. melanogaster was used to study genotoxicity of the medicinal plant Tabebuia impetiginosa. Lapachol (naphthoquinone) and beta-lapachone (quinone) are the two main chemical constituents of T. impetiginosa. These compounds have several biological properties. They induce apoptosis by generating oxygen-reactive species, thereby inhibiting topoisomerases (I and II) or inducing other enzymes dependent on NAD(P)H:quinone oxidoreductase 1, thus affecting cell cycle checkpoints. The SMART was used in the standard (ST) version, which has normal levels of cytochrome P450 (CYP) enzymes, to check the direct action of this compound, and in the high bioactivation (HB) version, which has a high constitutive level of CYP enzymes, to check for indirect action in three different T. impetiginosa concentrations (10%, 20% or 40% w/w). It was observed that T. impetiginosa alone did not modify the spontaneous frequencies of mutant spots in either cross. The negative results observed prompted us to study this phytotherapeuticum in association with the reference mutagen doxorubicin (DXR). In co-treated series, T. impetiginosa was toxic in both crosses at higher concentration, whereas in the HB cross, it induced a considerable potentiating effect (from ~24.0 to ~95.0%) on DXR genotoxity. Therefore, further research is needed to determine the possible risks associated with the exposure of living organisms to this complex mixture. PMID- 21637696 TI - Cytotoxic and genotoxic effects of Br-containing oxaphosphole on Allium cepa L. root tip cells and mouse bone marrow cells. AB - The continuous production and release of chemicals into the environment has led to the need to assess their genotoxicity. Numerous organophosphorus compounds with different structures have been synthesized in recent years, and several oxaphosphole derivatives are known to possess biological activity. Such chemical compounds may influence proliferating cells and cause disturbances of the genetic material. In this study, we examined the cytotoxicity and genotoxicity of 4-bromo N,N-diethyl-5,5-dimethyl-2,5-dihydro-1,2-oxaphosphol-2-amine 2-oxide (Br-oxph). In A. cepa cells, Br-oxph (10(-9) M, 10 (-6) M and 10 (-3) M) reduced the mitotic index 48 h after treatment with the two highest concentrations, with no significant effect at earlier intervals. Mitotic cells showed abnormalities 24 h and 48 h after treatment with the two lowest concentrations but there were no consistent changes in interphase cells. Bone marrow cells from mice treated with Br-oxph (2.82 x 10 (-3) MUg/kg) also showed a reduced mitotic index after 48 h and a greater percentage of cells with aberrations (principally chromatid and isochromatid breaks). These findings indicate the cytotoxicity and genotoxicity of Br-oxph in the two systems studied. PMID- 21637697 TI - Evaluation of genotoxicity using the micronucleus assay and nuclear abnormalities in the tropical sea fish Bathygobius soporator (Valenciennes, 1837) (Teleostei, Gobiidae). AB - The micronucleus and nuclear abnormalities assays have been used increasingly to evaluate genotoxicity of many compounds in polluted aquatic ecossystems. The aim of this study is to verify the efficiency of the micronucleus assay and nuclear abnormality assay in field and laboratory work, when using erythrocytes of the tropical marine fish Bathygobius soporator as genotoxicity biomarkers. Gill peripheral blood samples were obtained from specimens of Bathygobius soporator. In order to investigate the frequencies of micronuclei and to assess the sensitivity of species, the results were compared with samples taken at the reference site and maintained in the laboratory, and fish treated with cyclophosphamide. The micronucleus assay was efficient in demonstrating field pollution and reproducing results in the labotatory. There were significant higher frequencies of micronuclei in two sites subject to discharge of urban and industrial effluents. The nuclear abnormality assay did not appear to be an efficient tool for genotoxicity evaluation when compared with field samples taken at a reference site in laboratory, with a positive control. PMID- 21637698 TI - Expression of Msx1 and Dlx1 during Dumbo rat head development: Correlation with morphological features. AB - The Dumbo rat possesses some characteristics that evoke several human syndromes, such as Treacher-Collins: shortness of the maxillary, zygomatic and mandibular bones, and low position of the ears. Knowing that many homeobox genes are candidates in craniofacial development, we investigated the involvement of the Msx1 and Dlx1 genes in the Dumbo phenotype with the aim of understanding their possible role in abnormal craniofacial morphogenesis and examining the possibility of using Dumbo rat as an experimental model for understanding abnormal craniofacial development. We studied the expression of these genes during craniofacial morphogenesis by RT-PCR method. We used Dumbo embryos at E12 and E14 and included the Wistar strain as a control. Semi-quantitative PCR analysis demonstrated that Msx1 and Dlx1 are expressed differently between Dumbo and Wistar rats, indicating that their low expression may underly the Dumbo phenotype. PMID- 21637699 TI - Identification and phylogenetic inferences on stocks of sharks affected by the fishing industry off the Northern coast of Brazil. AB - The ongoing decline in abundance and diversity of shark stocks, primarily due to uncontrolled fishery exploitation, is a worldwide problem. An additional problem for the development of conservation and management programmes is the identification of species diversity within a given area, given the morphological similarities among shark species, and the typical disembarkation of processed carcasses which are almost impossible to differentiate. The main aim of the present study was to identify those shark species being exploited off northern Brazil, by using the 12S-16S molecular marker. For this, DNA sequences were obtained from 122 specimens collected on the docks and the fish market in Braganca, in the Brazilian state of Para. We identified at least 11 species. Three-quarters of the specimens collected were either Carcharhinus porosus or Rhizoprionodon sp, while a notable absence was the daggernose shark, Isogomphodon oxyrhyncus, previously one of the most common species in local catches. The study emphasises the value of molecular techniques for the identification of cryptic shark species, and the potential of the 12S-16S marker as a tool for phylogenetic inferences in a study of elasmobranchs. PMID- 21637700 TI - Evidence of two lineages of the dengue vector Aedes aegypti in the Brazilian Amazon, based on mitochondrial DNA ND4 gene sequences. AB - Genetic variation was estimated in ten samples populations of Aedes aegypti from the Brazilian Amazon, by using a 380 bp fragment of the mitochocondrial NADH dehydrogenase subunit 4 (ND4) gene. A total of 123 individuals were analyzed, whereby 13 haplotypes were found. Mean genetic diversity was slightly high (h = 0.666 +/- 0.029; pi = 0.0115 +/- 0.0010). Two AMOVA analyses indicated that most of the variation (~70%-72%) occurred within populations. The variation found among and between populations within the groups disclosed lower, but even so, highly significant values. F(ST) values were not significant in most of the comparisons, except for the samples from Pacaraima and Rio Branco. The isolation by distance (IBD) model was not significant (r = 0.2880; p = 0.097) when the samples from Pacaraima and Rio Branco were excluded from the analyses, this indicating that genetic distance is not related to geographic distance. This result may be explained either by passive dispersal patterns (via human migrations and commercial exchange) or be due to the recent expansion of this mosquito in the Brazilian Amazon. Phylogenetic relationship analysis showed two genetically distinct groups (lineages) within the Brazilian Amazon, each sharing haplotypes with populations from West Africa and Asia. PMID- 21637701 TI - Significant genetic differentiation among populations of Anomalocardia brasiliana (Gmelin, 1791): A bivalve with planktonic larval dispersion. AB - Four Brazilian populations of Anomalocardia brasiliana were tested for mutual genetic homogeneity, using data from 123 sequences of the mtDNA cytochrome oxidase c subunit I gene. A total of 36 haplotypes were identified, those shared being H3 (Canela Island, Prainha and Acupe) and both H5 and H9 (Prainha and Acupe). Haplotype diversity values were high, except for the Camurupim population, whereas nucleotide values were low in all the populations, except for that of Acupe. Only the Prainha population showed a deviation from neutrality and the SSD test did not reject the demographic expansion hypothesis. Fst values showed that the Prainha and Acupe populations represent a single stock, whereas in both the Canela Island and Camurupim stocks, population structures are different and independent. The observed structure at Canela Island may be due to the geographic distance between this population and the remainder. The Camurupim population does not share any haplotype with the remaining populations in northeastern Brazil. The apparent isolation could be due to the rocky barrier located facing the mouth of the Mamanguape River. The results highlight the importance of wide-scale studies to identify and conserve local genetic diversity, especially where migration is restricted. PMID- 21637702 TI - Allozyme comparison of two populations of Rineloricaria (Siluriformes, Loricariidae) from the Ivai River, upper Parana River basin, Brazil. AB - Two allopatric morphotypes of the genus Rinelocaria were compared through the allozyme electrophoresis technique: one morphotype, R. pentamaculata, from the Keller River in the middle stretch of the Ivai River basin and the other, R. aff. pentamaculata, from the Sao Joao River in the upper portion of the Ivai River basin. The morphotype from the Sao Joao River was collected upstream from the Sao Joao waterfall, which is about 80 m deep. Twelve enzymatic systems (AAT, ADH, EST, GCDH, G3PDH, GPI, IDH, LDH, MDH, ME, PGM and SOD) were analyzed, which allowed to score 22 loci. Only lociAat-2, Est-3 and Mdh-C showed polymorphism. The two samples differed in allele frequencies at the three polymorphic loci. The average expected heterozygosity for all loci was 0.0806 +/- 0.0447 in the Keller River sample. For the Sao Joao River morphotype, this value was 0.0489 +/- 0.0350. Nei' s genetic identity and distance between the two populations were respectively 0.9789 and 0.0213. Wright's F(IS) , F(IT) and F(ST) over all loci were estimated as 0.3121, 0.4021 and 0.1309, respectively. We consider that the two morphotypes represent species in statunascendi. PMID- 21637703 TI - Care transitions for older patients with musculoskeletal disorders: continuity from the providers' perspective. AB - INTRODUCTION: Care transitions are a common and frequently adverse aspect of health care, resulting in a high-risk period for both care quality and patient safety. Patients who have complex care needs and undergo treatment in multiple care settings, such as older patients with musculoskeletal disorders, may be at higher risk for poor care transitions. METHODS: Key informant interviews were used to gather in-depth information on transitional care issues, particularly those which impact informational continuity, from the perspective of a range of health professionals (eta=17) in care settings relevant to the care continuum of older patients with hip fractures. RESULTS: Three transitional care themes were identified; medical complexity impacts care trajectories, larger circles of care can be both beneficial and challenging, and a variety of channels and modes are required for meaningful information exchange. Many issues cut across each care setting, and address challenges to informational continuity among and between health care providers, patients, and caregivers. CONCLUSIONS: Medical complexity enlarges the circle of care which challenges care continuity. There may be fundamental elements which, regardless of care setting, strengthen transitional care quality. Standardized transitional care processes might help to offset informational discontinuity across care settings as a result of this population's larger circles of care. PMID- 21637704 TI - A new model of care collaboration for community-dwelling elders: findings and lessons learned from the NORC-Health Care linkage evaluation. AB - INTRODUCTION AND BACKGROUND: Few financial incentives in the United States encourage coordination across the health and social care systems. Supportive Service Programs (SSPs), operating in Naturally Occurring Retirement Communities (NORCs), attempt to increase access to care and enhance care quality for aging residents. This article presents findings from an evaluation conducted from 2004 to 2006 looking at the feasibility, quality and outcomes of linking health and social services through innovative NORC-SSP and health organization micro collaborations. METHODS: Four NORC-SSPs participated in the study by finding a health care organization or community-based physicians to collaborate with on addressing health conditions that could benefit from a biopsychosocial approach. Each site focused on a specific population, addressed a specific condition or problem, and created different linkages to address the target problem. Using a case study approach, incorporating both qualitative and quantitative methods, this evaluation sought to answer the following two primary questions: 1) Have the participating sites created viable linkages between their organizations that did not exist prior to the study; and, 2) To what extent have the linkages resulted in improvements in clinical and other health and social outcomes? RESULTS: FINDINGS SUGGEST THAT IMMEDIATE OUTCOMES WERE WIDELY ACHIEVED ACROSS SITES: knowledge of other sector providers' capabilities and services increased; communication across providers increased; identification of target population increased; and, awareness of risks, symptoms and health seeking behaviors among clients/patients increased. Furthermore, intermediate outcomes were also widely achieved: shared care planning, continuity of care, disease management and self care among clients improved. Evidence of improvements in distal outcomes was also found. DISCUSSION: Using simple, familiar and relatively low-tech approaches to sharing critical patient information among collaborating organizations, inter sector linkages were successfully established at all four sites. Seven critical success factors emerged that increase the likelihood that linkages will be implemented, effective and sustained: 1) careful goal selection; 2) meaningful collaboration; 3) appropriate role for patients/clients; 4) realistic interventions; 5) realistic expectations for implementation environment; 6) continuous focus on outcomes; and, 7) stable leadership. Focused, micro-level collaborations have the potential to improve care, increasing the chance that organizations will undertake such endeavors. PMID- 21637705 TI - Integrated care in Norway: the state of affairs years after regulation by law. AB - INTRODUCTION: A mandatory multidisciplinary plan for individual care, the 'Individual care Plan', was introduced by law in Norway in 2001. The regulation was established to meet the need for improved efficiency and quality of health and social services, and to increase patient involvement. The plan was intended for patients with long-term and complex needs for coordinated care. The aim of this study was to elaborate on knowledge of such planning processes in Norwegian municipalities. METHOD: A piloted questionnaire was sent to 92 randomly selected municipalities in 2005-2006, addressing local organization and participation in the work with individual care plans. Local political governance, size of the population, funds available for health care, and problems related to living conditions were indicators for analysing the extent to which the individual care plan was used five years after the regulation was introduced. RESULTS: Our results showed that 0.5% as opposed to an expected 3% of the population had an individual care plan. This was independent of the political, social and financial situation in the municipalities or the way the planning process had been carried out. The planning process was mostly taken care of by local health and social care professionals, rather than by hospital staff and general practitioners. DISCUSSION AND CONCLUSION: The low number of care plans and the oblique responsibility among professionals for planning showed that the objectives of the national initiative had not been achieved. More research is needed to determine the reasons for this lack of success and to contribute to solutions for improved multidisciplinary cooperation. PMID- 21637706 TI - Structuring an integrated care system: interpreted through the enacted diversity of the actors involved-the case of a French healthcare network. AB - RESEARCH QUESTION: We are looking at the process of structuring an integrated care system as an innovative process that swings back and forth between the diversity of the actors involved, local aspirations and national and regional regulations. We believe that innovation is enriched by the variety of the actors involved, but may also be blocked or disrupted by that diversity. Our research aims to add to other research, which, when questioning these integrated systems, analyses how the actors involved deal with diversity without really questioning it. CASE STUDY: The empirical basis of the paper is provided by case study analysis. The studied integrated care system is a French healthcare network that brings together healthcare professionals and various organisations in order to improve the way in which interventions are coordinated and formalised, in order to promote better detection and diagnosis procedures and the implementation of a care protocol. We consider this case as instrumental in developing theoretical proposals for structuring an integrated care system in light of the diversity of the actors involved. RESULTS AND DISCUSSION: We are proposing a model for structuring an integrated care system in light of the enacted diversity of the actors involved. This model is based on three factors: the diversity enacted by the leaders, three stances for considering the contribution made by diversity in the structuring process and the specific leading role played by those in charge of the structuring process. Through this process, they determined how the actors involved in the project were differentiated, and on what basis those actors were involved. By mobilising enacted diversity, the leaders are seeking to channel the emergence of a network in light of their own representation of that network. This model adds to published research on the structuring of integrated care systems. PMID- 21637707 TI - Integrated care requires integrated supervision. AB - INTRODUCTION: Given recent developments in integrated care, it is becoming increasingly important for the Dutch Health Care Inspectorate to direct its supervision in a way that may help speed up the implementation of integrated care. DESCRIPTION OF CARE PRACTICE: Since the implementation of integrated care for chronic patients is facing obstacles, alternative methods are required to ensure that the implementation process does not run into any delays. By applying a risk-based approach to integrated care providers, the Inspectorate can analyse the care providers' performance by means of quality indicators and rank them. In order to be effective, appropriated supervision arrangements will be applied to the care providers of integrated care. DISCUSSION: With a ranking model transparency will be improved and this may encourage integrated care providers to strive for greater quality due to the competition inherent in the system. Supervision based on advice and encouragement might be helpful in the implementation of integrated care. CONCLUSION: Integrated care also requires integrated supervision, which means the Inspectorate may have to reconsider its working methods and the composition of its inspection teams. PMID- 21637708 TI - Use of a knowledge synthesis by decision makers and planners to facilitate system level integration in a large Canadian provincial health authority. AB - PURPOSE: The study is an examination of how a knowledge synthesis, conducted to fill an information gap identified by decision makers and planners responsible for integrating health systems in a western Canadian health authority, is being used within that organization. METHODS: Purposive sampling and snowball technique were used to identify 13 participants who were interviewed about how they are using the knowledge synthesis for health services planning and decision-making. RESULTS: The knowledge synthesis is used by those involved in the strategic direction of the provincial healthcare organization and those tasked with the operationalization of integration at the provincial or local level. Both groups most frequently use the 10 key principles for integration, followed by the sections on integration processes, strategies and models. The key principles facilitate discussion on priority areas to be considered and provide a reference point for a desired future state. Perceived information gaps relate to a lack of detail on 'how to' strategies, tools and processes that would lead to successful integration. DISCUSSION AND CONCLUSION: The current project demonstrates that decision makers and planners will effectively use a knowledge synthesis if it is timely, relevant and accessible. The information can be applied at strategic and operations levels. Attention needs to be paid to include more information on implementation strategies and processes. Including knowledge users in identifying research questions will increase information uptake. PMID- 21637709 TI - Development of a program for tele-rehabilitation of COPD patients across sectors: co-innovation in a network. AB - INTRODUCTION: The aim of the Telekat project is to prevent re-admissions of patients with chronic obstructive pulmonary disease (COPD) by developing a preventive program of tele-rehabilitation across sectors for COPD patients. The development of the program is based on a co-innovation process between COPD patients, relatives, healthcare professionals and representatives from private firms and universities. This paper discusses the obstacles that arise in the co innovation process of developing an integrated technique for tele-rehabilitation of COPD patients. THEORY: Network and innovation theory. METHODS: The case study was applied. A triangulation of data collection techniques was used: documents, observations (123 hours), qualitative interviews (n=32) and action research. FINDINGS: Obstacles were identified in the network context; these obstacles included the mindset of the healthcare professionals, inter-professionals relations, views of technology as a tool and competing visions for the goals of tele-rehabilitation. CONCLUSION: We have identified obstacles that emerge in the co-innovation process when developing a programme for tele-rehabilitation of COPD patients in an inter-organizational context. Action research has been carried out and can have helped to facilitate the co-innovation process. PMID- 21637710 TI - Adopting telehealth as a tool of integrated care: what type of research is required to justify the investment? PMID- 21637711 TI - Inhibitory actions of Anti-Mullerian Hormone (AMH) on ovarian primordial follicle assembly. AB - The current study was designed to investigate the actions of Anti-Mullerian Hormone (AMH) on primordial follicle assembly. Ovarian primordial follicles develop from the breakdown of oocyte nests during fetal development for the human and immediately after birth in rodents. AMH was found to inhibit primordial follicle assembly and decrease the initial primordial follicle pool size in a rat ovarian organ culture. The AMH expression was found to be primarily in the stromal tissue of the ovaries at this period of development, suggesting a stromal epithelial cell interaction for primordial follicle assembly. AMH was found to promote alterations in the ovarian transcriptome during primordial follicle assembly with over 200 genes with altered expression. A gene network was identified suggesting a potential central role for the Fgf2/Nudt6 antisense transcript in the follicle assembly process. A number of signal transduction pathways are regulated by AMH actions on the ovarian transcriptome, in particular the transforming growth factor-beta (TGFbeta) signaling process. AMH is the first hormone/protein shown to have an inhibitory action on primordial follicle assembly. Due to the critical role of the primordial follicle pool size for female reproduction, elucidation of factors, such as AMH, that regulate the assembly process will provide insights into potential therapeutics to manipulate the pool size and female reproduction. PMID- 21637712 TI - High throughput ratio imaging to profile caspase activity: potential application in multiparameter high content apoptosis analysis and drug screening. AB - Recent advancement in the area of green fluorescent protein techniques coupled with microscopic imaging has significantly contributed in defining and dissecting subcellular changes of apoptosis with high spatio-temporal resolution. Although single cell based studies using EGFP and associated techniques have provided valuable information of initiation and hierarchical changes of apoptosis, they are yet to be exploited for multiparameter cell based real time analysis for possible drug screening or pathway defining in a high throughput manner. Here we have developed multiple cancer cell lines expressing FRET sensors for active caspases and adapted them for high throughput live cell ratio imaging, enabling high content image based multiparameter analysis. Sensitivity of the system to detect live cell caspase activation was substantiated by confocal acceptor bleaching as well as wide field FRET imaging. Multiple caspase-specific activities of DEVDase, IETDase and LEHDase were analysed simultaneously with other decisive events of cell death. Through simultaneous analysis of caspase activation by FRET ratio change coupled with detection of mitochondrial membrane potential loss or superoxide generation, we identified several antitumor agents that induced caspase activation with or without membrane potential loss or superoxide generation. Also, cells that escaped the initial drug-induced caspase activation could be easily followed up for defining long term fate. Employing such a revisit imaging strategy of the same area, we have tracked the caspase surviving fractions with multiple drugs and its subsequent response to retreatment, revealing drug-dependent diverging fate of surviving cells. This thereby indicates towards a complex control of drug induced tumor resistance. The technique described here has wider application in both screening of compound libraries as well as in defining apoptotic pathways by linking multiple signaling to identify non-classical apoptosis inducing agents, the greatest advantage being that the high content information obtained are from individual cells rather than being population based. PMID- 21637713 TI - Lack of the long pentraxin PTX3 promotes autoimmune lung disease but not glomerulonephritis in murine systemic lupus erythematosus. AB - The long pentraxin PTX3 has multiple roles in innate immunity. For example, PTX3 regulates C1q binding to pathogens and dead cells and regulates their uptake by phagocytes. It also inhibits P-selectin-mediated recruitment of leukocytes. Both of these mechanisms are known to be involved in autoimmunity and autoimmune tissue injury, e.g. in systemic lupus erythematosus, but a contribution of PTX3 is hypothetical. To evaluate a potential immunoregulatory role of PTX3 in autoimmunity we crossed Ptx3-deficient mice with Fas-deficient (lpr) C57BL/6 (B6) mice with mild lupus-like autoimmunity. PTX3 was found to be increasingly expressed in kidneys and lungs of B6lpr along disease progression. Lack of PTX3 impaired the phagocytic uptake of apoptotic T cells into peritoneal macrophages and selectively expanded CD4/CD8 double negative T cells while other immune cell subsets and lupus autoantibody production remained unaffected. Lack of PTX3 also aggravated autoimmune lung disease, i.e. peribronchial and perivascular CD3+ T cell and macrophage infiltrates of B6lpr mice. In contrast, histomorphological and functional parameters of lupus nephritis remained unaffected by the Ptx3 genotype. Together, PTX3 specifically suppresses autoimmune lung disease that is associated with systemic lupus erythematosus. Vice versa, loss-of-function mutations in the Ptx3 gene might represent a genetic risk factor for pulmonary (but not renal) manifestations of systemic lupus or other autoimmune diseases. PMID- 21637714 TI - Threshold learning dynamics in social networks. AB - Social learning is defined as the ability of a population to aggregate information, a process which must crucially depend on the mechanisms of social interaction. Consumers choosing which product to buy, or voters deciding which option to take with respect to an important issue, typically confront external signals to the information gathered from their contacts. Economic models typically predict that correct social learning occurs in large populations unless some individuals display unbounded influence. We challenge this conclusion by showing that an intuitive threshold process of individual adjustment does not always lead to such social learning. We find, specifically, that three generic regimes exist separated by sharp discontinuous transitions. And only in one of them, where the threshold is within a suitable intermediate range, the population learns the correct information. In the other two, where the threshold is either too high or too low, the system either freezes or enters into persistent flux, respectively. These regimes are generally observed in different social networks (both complex or regular), but limited interaction is found to promote correct learning by enlarging the parameter region where it occurs. PMID- 21637715 TI - Detailed analysis of variants in FTO in association with body composition in a cohort of 70-year-olds suggests a weakened effect among elderly. AB - BACKGROUND: The rs9939609 single-nucleotide polymorphism (SNP) in the fat mass and obesity (FTO) gene has previously been associated with higher BMI levels in children and young adults. In contrast, this association was not found in elderly men. BMI is a measure of overweight in relation to the individuals' height, but offers no insight into the regional body fat composition or distribution. OBJECTIVE: To examine whether the FTO gene is associated with overweight and body composition-related phenotypes rather than BMI, we measured waist circumference, total fat mass, trunk fat mass, leg fat mass, visceral and subcutaneous adipose tissue, and daily energy intake in 985 humans (493 women) at the age of 70 years. In total, 733 SNPs located in the FTO gene were genotyped in order to examine whether rs9939609 alone or the other SNPs, or their combinations, are linked to obesity-related measures in elderly humans. DESIGN: Cross-sectional analysis of the Prospective Investigation of the Vasculature in Uppsala Seniors (PIVUS) cohort. RESULTS: Neither a single SNP, such as rs9939609, nor a SNP combination was significantly linked to overweight, body composition-related measures, or daily energy intake in elderly humans. Of note, these observations hold both among men and women. CONCLUSIONS: Due to the diversity of measurements included in the study, our findings strengthen the view that the effect of FTO on body composition appears to be less profound in later life compared to younger ages and that this is seemingly independent of gender. PMID- 21637716 TI - Co-variation of tonality in the music and speech of different cultures. AB - Whereas the use of discrete pitch intervals is characteristic of most musical traditions, the size of the intervals and the way in which they are used is culturally specific. Here we examine the hypothesis that these differences arise because of a link between the tonal characteristics of a culture's music and its speech. We tested this idea by comparing pitch intervals in the traditional music of three tone language cultures (Chinese, Thai and Vietnamese) and three non-tone language cultures (American, French and German) with pitch intervals between voiced speech segments. Changes in pitch direction occur more frequently and pitch intervals are larger in the music of tone compared to non-tone language cultures. More frequent changes in pitch direction and larger pitch intervals are also apparent in the speech of tone compared to non-tone language cultures. These observations suggest that the different tonal preferences apparent in music across cultures are closely related to the differences in the tonal characteristics of voiced speech. PMID- 21637717 TI - Differential effects of Bartonella henselae on human and feline macro- and micro vascular endothelial cells. AB - Bartonella henselae, a zoonotic agent, induces tumors of endothelial cells (ECs), namely bacillary angiomatosis and peliosis in immunosuppressed humans but not in cats. In vitro studies on ECs represent to date the only way to explore the interactions between Bartonella henselae and vascular endothelium. However, no comparative study of the interactions between Bartonella henselae and human (incidental host) ECs vs feline (reservoir host) ECs has been carried out because of the absence of any available feline endothelial cell lines.To this purpose, we have developed nine feline EC lines which allowed comparing the effects of Bartonella strains on human and feline micro-vascular ECs representative of the infection development sites such as skin, versus macro-vascular ECs, such as umbilical vein.Our model revealed intrinsic differences between human (Human Skin Microvascular ECs -HSkMEC and Human Umbilical Vein ECs - iHUVEC) and feline ECs susceptibility to Bartonella henselae infection.While no effect was observed on the feline ECs upon Bartonella henselae infection, the human ones displayed accelerated angiogenesis and wound healing.Noticeable differences were demonstrated between human micro- and macro-vasculature derived ECs both in terms of pseudo-tube formation and healing. Interestingly, Bartonella henselae effects on human ECs were also elicited by soluble factors.Neither Bartonella henselae infected Human Skin Microvascular ECs clinically involved in bacillary angiomatosis, nor feline ECs increased cAMP production, as opposed to HUVEC.Bartonella henselae could stimulate the activation of Vascular Endothelial Growth Factor Receptor-2 (VEGFR-2) in homologous cellular systems and trigger VEGF production by HSkMECs only, but not iHUVEC or any feline ECs tested.These results may explain the decreased pathogenic potential of Bartonella henselae infection for cats as compared to humans and strongly suggest that an autocrine secretion of VEGF by human skin endothelial cells might induce their growth and ultimately lead to bacillary angiomatosis formation. PMID- 21637718 TI - Peripheral vascular dysfunction in chronic kidney disease. AB - There is an increased prevalence of cardiovascular disease- (CVD-) related mortality in patients with chronic kidney disease (CKD). Endothelial dysfunction is a primary event in the development of atherosclerosis and hypertension and likely contributes to the elevated cardiovascular risk in CKD. Endothelial dysfunction has been shown to occur in the peripheral vasculature of patients with both severe and moderate CKD. Mechanisms include oxidative stress, L arginine deficiency, and elevated plasma levels of ADMA. Interventions designed to restore vascular function in patients with CKD have shown mixed results. Evidence from cell culture studies suggest that the accumulation of uremic toxins inhibits L-arginine transport and reduces nitric oxide production. The results of these studies suggest that endothelial dysfunction may become less reversible with advancing kidney disease. The purpose of this paper is to present the current literature pertaining to potential mechanisms of peripheral vascular dysfunction in chronic kidney disease and to identify possible targets for treatment. PMID- 21637719 TI - Salmonella enteritidis Infection Complicated by Acute Myocarditis: A Case Report and Review of the Literature. AB - Salmonella spp. is the cause of commonly encountered infections, with seasonal pattern of occurrence and worldwide distribution. Some of the clinical manifestations such as gastroenteritis and bacteremia are common, whereas others like mycotic aneurysms and osteomyelitis are infrequent especially in immunocompetent patients. Salmonella has been rarely described as a cause of myocarditis in the literature. We describe a case of an 18-year-old previously healthy male patient with myocarditis after Salmonella enteritidis infection. Clinical manifestations and diagnostic approach of this severe complication are discussed with a review of the literature. PMID- 21637720 TI - Electrocardiographic and electrophysiologic characteristics of ventricular extrasystoles arising from the aortomitral continuity. AB - Left ventricular outflow tract arrhythmias originating from the aortomitral continuity, the left coronary cusp, the superior basal septum, and the epicardial left ventricular summit display common electrocardiographic and electrophysiological features, probably due to the close proximity of those locations. Catheter ablation of these arrhythmias can be challenging. The case of a 68-year-old male with frequent premature ventricular extrasystoles arising from the aortomitral continuity of the basal left ventricle is described. The electrocardiographic and electrophysiologic characteristics of this arrhythmia are discussed. PMID- 21637721 TI - Effects of Phonotactic Probabilities on the Processing of Spoken Words and Nonwords by Adults with Cochlear Implants Who Were Postlingually Deafened. AB - Probabilistic phonotactics refers to the frequency with which segments and sequences of segments occur in syllables and words. Knowledge of phonotactics has been shown to be an important source of information in recognizing spoken words in listeners with normal hearing. Two online tasks (an auditory same-different task and an auditory lexical decision task) were used to examine the use of phonotactic information by adults who were postlingually deafened who have received cochlear implants. The results of the experiments showed that cochlear implant patients with better word recognition abilities (as measured by the Northwestern University Auditory Test No. 6 (NU-6) produced patterns of results that were similar to the pattern of results obtained from listeners with normal hearing in Vitevitch and Luce (1999). This finding suggests that cochlear implant patients with better word recognition abilities use lexical and sublexical representations to process spoken words, much like listeners with normal hearing. In contrast, cochlear implant patients with poor word recognition abilities could not differentiate between stimuli varying in phonotactic probability and lexicality, suggesting that less distinct representations are used by these patients to process spoken words. The implications of these results for outcome assessments and clinical interventions are discussed. PMID- 21637722 TI - SMOKE-FREE ORDINANCES INCREASE RESTAURANT PROFIT AND VALUE. AB - This study estimates the economic value added to a restaurant by a smoke-free policy using regression analysis of the purchase price of restaurants, as a function of the presence of a smoke-free law and other control variables. There was a median increase of 16% (interquartile range 11% to 25%) in the sale price of a restaurant in a jurisdiction with a smoke-free law compared to a comparable restaurant in a community without such a law. This result indicates that, contrary to claims made by the tobacco industry and other opponents of smoke-free laws, these laws are associated with an increase in restaurant profitability. PMID- 21637723 TI - The Presentation of Narcissistic Personality Disorder in an Octogenarian: Converging Evidence from Multiple Sources. AB - Little is known about personality disorders (PDs) in later life. One reason for this dearth of knowledge is that many investigators believe that PDs soften with age. Recent anecdotal and empirical evidence, however, suggests that PDs are still very relevant in later life and may actually have unique presentations and consequences. The DSM-IV PD criteria seem to overlook these possibilities, perhaps because the personalities of older adults were not sufficiently understood when these criteria were written. But without age-appropriate criteria, clinicians and investigators who work with older adults may be unable to measure PDs adequately in their clients and research participants. A starting point for better understanding these disorders in older adults is the presentation of rich, empirical, clinical descriptions of symptoms and related behaviors using data from multiple instruments and sources. To this end, we describe in depth a case of narcissistic PD (NPD) in a woman in her mid 80s. This case study reveals that NPD is indeed relevant in the context of later life and impairs functioning in significant ways. PMID- 21637724 TI - Identification of Multivariate Responders/Non-Responders Using Bayesian Growth Curve Latent Class Models. AB - In this paper, we propose a multivariate growth curve mixture model that groups subjects based on multiple symptoms measured repeatedly over time. Our model synthesizes features of two models. First, we follow Roy and Lin (2000) in relating the multiple symptoms at each time point to a single latent variable. Second, we use the growth mixture model of Muthen and Shedden (1999) to group subjects based on distinctive longitudinal profiles of this latent variable. The mean growth curve for the latent variable in each class defines that class's features. For example, a class of "responders" would have a decline in the latent symptom summary variable over time. A Bayesian approach to estimation is employed where the methods of Elliott et al (2005) are extended to simultaneously estimate the posterior distributions of the parameters from the latent variable and growth curve mixture portions of the model. We apply our model to data from a randomized clinical trial evaluating the efficacy of Bacillus Calmette-Guerin (BCG) in treating symptoms of Interstitial Cystitis. In contrast to conventional approaches using a single subjective Global Response Assessment, we use the multivariate symptom data to identify a class of subjects where treatment demonstrates effectiveness. Simulations are used to confirm identifiability results and evaluate the performance of our algorithm. The definitive version of this paper is available at onlinelibrary.wiley.com. PMID- 21637725 TI - Elastomeric polypeptide-based biomaterials. AB - Elastomeric proteins are characterized by their large extensibility before rupture, reversible deformation without loss of energy, and high resilience upon stretching. Motivated by their unique mechanical properties, there has been tremendous research in understanding and manipulating elastomeric polypeptides, with most work conducted on the elastins but more recent work on an expanded set of polypeptide elastomers. Facilitated by biosynthetic strategies, it has been possible to manipulate the physical properties, conformation, and mechanical properties of these materials. Detailed understanding of the roles and organization of the natural structural proteins has permitted the design of elastomeric materials with engineered properties, and has thus expanded the scope of applications from elucidation of the mechanisms of elasticity to the development of advanced drug delivery systems and tissue engineering substrates. PMID- 21637726 TI - Microscale 1-3-Type (Na,K)NbO(3)-Based Pb-Free Piezocomposites for High-Frequency Ultrasonic Transducer Applications. AB - Fine-grained Pb-free (Na(0.535)K(0.485))(0.95)Li(0.05)(Nb(0.8)Ta(0.2))O(3) (NKLNT) piezoceramics prepared by spark plasma sintering (SPS) technique was used to fabricate NKLNT/epoxy 1-3 composites with a modified dice-fill method. Because of its good machinability, SPSed NKLNT ceramic rods could be miniaturized to a lateral width of 50 um. After lapping down to 56 um in thickness, the composite was used to fabricate an ultrasonic transducer as the active piezoelectric element. This composite transducer showed a bandwidth at -6 dB nearly 90%at a center frequency of 29 MHz, demonstrating that this Pb-free composite thick film is very promising for the fabrication of high-frequency ultrasonic transducers in medical imaging applications. PMID- 21637727 TI - The Relations Among Oral and Silent Reading Fluency and Comprehension in Middle School: Implications for Identification and Instruction of Students With Reading Difficulties. AB - The purpose of this study was to investigate the relations among oral and silent reading fluency and reading comprehension for students in Grades 6 to 8 (n = 1,421) and the use of fluency scores to identify middle school students who are at risk for failure on a high-stakes reading test. Results indicated moderate positive relations between measures of fluency and comprehension. Oral reading fluency (ORF) on passages was more strongly related to reading comprehension than ORF on word lists. A group-administered silent reading sentence verification test approximated the classification accuracy of individually administered ORF passages. The correlation between a maze task and comprehension was weaker than has been reported for elementary students. The best predictor of a high-stakes reading comprehension test was the previous year's administration of the grade appropriate test; fluency and verbal knowledge measures accounted for only small amounts of unique variance beyond that accounted for by the previous year's administration. PMID- 21637728 TI - Effects of Speaking Rate on Word Recognition in Parkinson's Disease and Normal Aging. AB - Current theories of basal ganglia function emphasize their role in the integration of sensory information into motor activities, particularly in the control of movement timing. People with basal ganglia disorders such as Parkinson's disease exhibit poor temporal control of movements, in general and articulation in particular, as demonstrated by irregular speaking rate, reduced stress contrasts, and reduced movement durations and velocities. Previous research has implicated sensory deficits as contributory factors in limb movement control in patients with Parkinson's disease; however, the relation between sensory deficits and speech-movement abnormalities has not been documented. In the present study, the existence of perceptual processing difficulties of speaking rate was investigated in subjects with Parkinsonian dysarthria (PD). Comparisons in perception were made between subjects with PD, neurologically normal geriatrics (GN) and neurologically normal young adults (YN) for accuracy in identification of words presented at different speaking rates. We hypothesized that word-identification scores would be lower for PD and GN subjects compared to the YN subjects, an effect that was supported by the data. We also expected that there would be differences between the GN and PD subjects in their accuracy of word identification at a faster speaking rate, an hypothesis that was not supported by the data. Rather, GN and PD subjects differed in identification scores for words spoken at a slow rate. PD subjects who had faster habitual speaking rates (HSR) had significantly lower word-identification scores in the slow compared to conversational rate conditions, a relation that was significant r = +0.64). These data suggest the need to consider perceptual deficits as an additional factor that contributes to rate variations in PD speech. PMID- 21637729 TI - Acceptance, Communication Mode and Use of Audio Computer-Assisted Self Interview Using Touchscreen to Identify Risk Factors among Pregnant Minority Women. AB - This paper evaluates the acceptability, communication mode and use of audio computer-assisted self-interview (A-CASI) among minority pregnant women receiving prenatal care in six Washington, DC sites. A total of 2,913 women were screened for demographic eligibility (18+ years old, <29 weeks gestation, Black/African American or Hispanic) and risk (smoking, environmental tobacco smoke exposure, depression, intimate partner violence). Questions were displayed on touch screen laptop monitors and heard through earphones. The mean length of time to complete the screener was almost 6 minutes.A-CASI experience, which included difficulty in using the computer, acceptability (enjoyment), and preferred communication mode, was compared across sites, the eligibility and risk groups and a subset of 878 enrolled women for whom educational attainment and receipt of WIC (a proxy for income) were available. Respondents thought A-CASI was not difficult to use and liked using the computer. Black/African-American or Hispanic respondents enjoyed it significantly more than did respondents of other race/ethnicities. Respondents who were demographically eligible, Black/African-American or Hispanic, or with lower education levels listened to questions significantly more than did their counterparts. Mainly listening or listening and reading does not impact burden in terms of the length of time it took to complete the screener.The acceptance of A CASI as a screening tool opens the door for more uses of this technology in health-related fields. The laptop computer and headphones provide privacy and mobility so the technology can be used to ask sensitive questions in almost any locale, including busy clinic settings. PMID- 21637730 TI - Ontological realism: A methodology for coordinated evolution of scientific ontologies. AB - Since 2002 we have been testing and refining a methodology for ontology development that is now being used by multiple groups of researchers in different life science domains. Gary Merrill, in a recent paper in this journal, describes some of the reasons why this methodology has been found attractive by researchers in the biological and biomedical sciences. At the same time he assails the methodology on philosophical grounds, focusing specifically on our recommendation that ontologies developed for scientific purposes should be constructed in such a way that their terms are seen as referring to what we call universals or types in reality. As we show, Merrill's critique is of little relevance to the success of our realist project, since it not only reveals no actual errors in our work but also criticizes views on universals that we do not in fact hold. However, it nonetheless provides us with a valuable opportunity to clarify the realist methodology, and to show how some of its principles are being applied, especially within the framework of the OBO (Open Biomedical Ontologies) Foundry initiative. PMID- 21637731 TI - The role of stem cells in tumor targeting and growth suppression of gliomas. AB - Glioma remains the most challenging solid organ tumor to treat successfully. Based on the capacity of stem cells to migrate extensively and target invading glioma cells, the transplantation of stem cells as a cell-based delivery system may provide additional tools for the treatment of gliomas. In addition to the use of modified stem cells for the delivery of therapeutic agents, unmodified stem cells have been shown to have growth-suppressing effects on tumors in vitro and in vivo. This review outlines the probable factors involved in tumor tropism and tumor growth suppression, with a specific focus on the use of unmodified stem cells in the treatment of gliomas. Based on these and further future data, clinical trials may be justified. PMID- 21637733 TI - Crystal methamphetamine use among American Indian and White youth in Appalachia: Social context, masculinity, and desistance. AB - Rural areas and American Indian reservations are hotspots for the use of crystal methamphetamine ("meth") in the United States, yet there is little ethnographic data describing meth use in these areas. This study draws upon three years of ethnographic work conducted with American Indian and White youth in Appalachia during the height of the meth epidemic. It describes how historical, cultural, and socioeconomic processes influence vulnerability to meth use in Appalachia, and highlights the role of social relationships and meaning-making in facilitating desistance and recovery from meth use. The first section shows how crystal meth filled a particular functional niche in the lives of many young men, alleviating boredom and anomie linked to recent socioeconomic changes and labor opportunities in the region, and intersecting with local understandings of masculinity and forms of military identity. Here, ethnographic and interview data converge to illustrate how social role expectations, recent socioeconomic change, and meth's pharmacological properties converge to create vulnerability to meth use in Appalachia. The second section draws upon two American Indian narratives of desistance. These youth described recently severed social relationships and acute feelings of social isolation during the initiation of meth use. Both also described dramatic close calls with death that facilitated their eventual desistance from use, involving repaired social relationships and the establishment of new lives and hope. These interviews illustrate how changes in social relationships were linked with both initiation and desistance from meth use, and how religious interpretations of near-death experiences structured narratives of cessation and redemption. PMID- 21637732 TI - Interactions of host proteins with the murine leukemia virus integrase. AB - Retroviral infections cause a variety of cancers in animals and a number of diverse diseases in humans such as leukemia and acquired immune deficiency syndrome. Productive and efficient proviral integration is critical for retroviral function and is the key step in establishing a stable and productive infection, as well as the mechanism by which host genes are activated in leukemogenesis. Host factors are widely anticipated to be involved in all stages of the retroviral life cycle, and the identification of integrase interacting factors has the potential to increase our understanding of mechanisms by which the incoming virus might appropriate cellular proteins to target and capture host DNA sequences. Identification of MoMLV integrase interacting host factors may be key to designing efficient and benign retroviral-based gene therapy vectors; key to understanding the basic mechanism of integration; and key in designing efficient integrase inhibitors. In this review, we discuss current progress in the field of MoMLV integrase interacting proteins and possible roles for these proteins in integration. PMID- 21637734 TI - Random Regression Models Based On The Skew Elliptically Contoured Distribution Assumptions With Applications To Longitudinal Data. AB - Bartolucci et al.(2003) extended the distribution assumption from the normal (Lyles et al., 2000) to the elliptical contoured distribution (ECD) for random regression models used in analysis of longitudinal data accounting for both undetectable values and informative drop-outs. In this paper, the random regression models are constructed on the multivariate skew ECD. A real data set is used to illustrate that the skew ECDs can fit some unimodal continuous data better than the Gaussian distributions or more general continuous symmetric distributions when the symmetric distribution assumption is violated. Also, a simulation study is done for illustrating the model fitness from a variety of skew ECDs. The software we used is SAS/STAT, V. 9.13. PMID- 21637735 TI - The Differential Impact on Children of Inter- and Intra-Community Violence in Northern Ireland. AB - This study explores distinctions in Northern Ireland between inter-community (i.e. sectarian) and intra-community (i.e. nonsectarian) violence and their respective impacts on children, and considers these forms of violence in relation to children's processes of emotional security about community conflict. Preliminary work was based on focus groups with mothers in Belfast, followed by a quantitative study involving mothers in Derry/Londonderry. Support emerged for a conceptually-based distinction between sectarian and nonsectarian violence and differential prediction of children's adjustment problems, which was more closely linked with sectarian than nonsectarian community violence. Pertinent to explanatory mechanisms, community violence, especially sectarian, related to mothers' perceptions of children's emotional insecurity about community. Findings are discussed in terms of future directions for understanding community violence and child development in cultural context. PMID- 21637736 TI - QUANTITATIVE MODELING OF SPATIO-TEMPORAL DYNAMICS OF INFERIOR OLIVE NEURONS WITH A SIMPLE CONDUCTANCE-BASED MODEL. AB - Inferior olive (IO) neurons project to the cerebellum and contribute to motor control. They can show intriguing spatio-temporal dynamics with rhythmic and synchronized spiking. IO neurons are connected to their neighbors via gap junctions to form an electrically coupled network, and so it is considered that this coupling contributes to the characteristic dynamics of this nucleus. Here, we demonstrate that a gap junction-coupled network composed of simple conductance based model neurons (a simplified version of a Hodgkin-Huxley type neuron) reproduce important aspects of IO activity. The simplified phenomenological model neuron facilitated the analysis of the single cell and network properties of the IO while still quantitatively reproducing the spiking patterns of complex spike activity observed by simultaneous recording in anesthetized rats. The results imply that both intrinsic bistability of each neuron and gap junction coupling among neurons play key roles in the generation of the spatio-temporal dynamics of IO neurons. PMID- 21637737 TI - Estimating Causal Effects in Trials Involving Multi-Treatment Arms Subject to Non compliance: A Bayesian framework. AB - Data analysis for randomized trials including multi-treatment arms is often complicated by subjects who do not comply with their treatment assignment. We discuss here methods of estimating treatment efficacy for randomized trials involving multi-treatment arms subject to non-compliance. One treatment effect of interest in the presence of non-compliance is the complier average causal effect (CACE) (Angrist et al. 1996), which is defined as the treatment effect for subjects who would comply regardless of the assigned treatment. Following the idea of principal stratification (Frangakis & Rubin 2002), we define principal compliance (Little et al. 2009) in trials with three treatment arms, extend CACE and define causal estimands of interest in this setting. In addition, we discuss structural assumptions needed for estimation of causal effects and the identifiability problem inherent in this setting from both a Bayesian and a classical statistical perspective. We propose a likelihood-based framework that models potential outcomes in this setting and a Bayes procedure for statistical inference. We compare our method with a method of moments approach proposed by Cheng & Small (2006) using a hypothetical data set, and further illustrate our approach with an application to a behavioral intervention study (Janevic et al. 2003). PMID- 21637738 TI - THE USE OF A DUAL-TASK PARADIGM FOR ASSESSING SPEECH INTELLIGIBILITY IN CLIENTS WITH PARKINSON DISEASE. AB - Differences in the clinical and ecological manifestations of reduced intelligibility for individuals with dysarthria related to Parkinson disease (PD) have been reported in the literature. The current study explored whether a dual task paradigm could be used during intelligibility testing to collect speech samples that were representative of functional performance. Intelligibility was calculated for four speakers with PD and four age-matched controls (CG) based on single-word, sentences, and monologue tasks recorded in single-and dual-task conditions and a spontaneous speech sample. In the dual-task condition, speakers produced the target speech sample and performed a simultaneous motor task, turning a nut on a bolt. No significant differences in intelligibility were found for the CG. For speakers with PD, differences between conditions were statistically significant for all speech tasks. Intelligibility scores in the dual-task condition were lower, with variability between tasks and speakers noted. There was a significant difference between scores for the monologue in the single-task condition and the spontaneous sample; however, there was no significant difference between the monologue in the dual-task condition and the spontaneous sample. Findings suggest that including a simple motor task during a clinical assessment may help elicit speech samples that are representative of a speaker's typical speech production. PMID- 21637739 TI - Supporting positive dimensions of health, challenges in mental health care. AB - This paper will explore two contrasting paradigms in mental health care and their relationship to evidence-based practice. The biomedical perspective of pathogenesis and the health perspective of salotogenesis are two major diverse views in mental health care. Positive dimensions of health are traditionally viewed as software not suitable for statistical analysis, while absence of symptoms of disease are regarded as measurable and suitable for statistical analysis and appropriate as a foundation of evidence-based practice. If the main goal of mental health care is to enhance subjectively experienced health among patients, it will not be sufficient to evaluate absence of symptoms of disease as a measure of quality of care. The discussion focuses on the paradox of evidence based absence of illness and disease versus subjectively experienced health and well-being as criterions of quality of care in mental health care. PMID- 21637740 TI - In what direction should we go to promote health in mental health care? AB - There is a growing recognition of the need for health promotion interventions in all health care today. In spite of this, health promotion interventions among patients with mental illnesses have been scarce in research, practice, and policies. There is also an ambiguous interpretation of the definition of health promotion in the literature. The emphasis in this paper is thus to (1) discuss why we should pay attention to the interpretations of the concept of health promotion and (2) present a possible model for what nurses do when they intend to promote health in mental health care. This paper was presented at the Nordic Conference of Mental Health Nursing in Helsinki, Finland in 2010. PMID- 21637741 TI - Speed/accuracy trade-off between the habitual and the goal-directed processes. AB - Instrumental responses are hypothesized to be of two kinds: habitual and goal directed, mediated by the sensorimotor and the associative cortico-basal ganglia circuits, respectively. The existence of the two heterogeneous associative learning mechanisms can be hypothesized to arise from the comparative advantages that they have at different stages of learning. In this paper, we assume that the goal-directed system is behaviourally flexible, but slow in choice selection. The habitual system, in contrast, is fast in responding, but inflexible in adapting its behavioural strategy to new conditions. Based on these assumptions and using the computational theory of reinforcement learning, we propose a normative model for arbitration between the two processes that makes an approximately optimal balance between search-time and accuracy in decision making. Behaviourally, the model can explain experimental evidence on behavioural sensitivity to outcome at the early stages of learning, but insensitivity at the later stages. It also explains that when two choices with equal incentive values are available concurrently, the behaviour remains outcome-sensitive, even after extensive training. Moreover, the model can explain choice reaction time variations during the course of learning, as well as the experimental observation that as the number of choices increases, the reaction time also increases. Neurobiologically, by assuming that phasic and tonic activities of midbrain dopamine neurons carry the reward prediction error and the average reward signals used by the model, respectively, the model predicts that whereas phasic dopamine indirectly affects behaviour through reinforcing stimulus-response associations, tonic dopamine can directly affect behaviour through manipulating the competition between the habitual and the goal-directed systems and thus, affect reaction time. PMID- 21637742 TI - How modeling can reconcile apparently discrepant experimental results: the case of pacemaking in dopaminergic neurons. AB - Midbrain dopaminergic neurons are endowed with endogenous slow pacemaking properties. In recent years, many different groups have studied the basis for this phenomenon, often with conflicting conclusions. In particular, the role of a slowly-inactivating L-type calcium channel in the depolarizing phase between spikes is controversial, and the analysis of slow oscillatory potential (SOP) recordings during the blockade of sodium channels has led to conflicting conclusions. Based on a minimal model of a dopaminergic neuron, our analysis suggests that the same experimental protocol may lead to drastically different observations in almost identical neurons. For example, complete L-type calcium channel blockade eliminates spontaneous firing or has almost no effect in two neurons differing by less than 1% in their maximal sodium conductance. The same prediction can be reproduced in a state of the art detailed model of a dopaminergic neuron. Some of these predictions are confirmed experimentally using single-cell recordings in brain slices. Our minimal model exhibits SOPs when sodium channels are blocked, these SOPs being uncorrelated with the spiking activity, as has been shown experimentally. We also show that block of a specific conductance (in this case, the SK conductance) can have a different effect on these two oscillatory behaviors (pacemaking and SOPs), despite the fact that they have the same initiating mechanism. These results highlight the fact that computational approaches, besides their well known confirmatory and predictive interests in neurophysiology, may also be useful to resolve apparent discrepancies between experimental results. PMID- 21637743 TI - Paced-mating increases the number of adult new born cells in the internal cellular (granular) layer of the accessory olfactory bulb. AB - The continuous production and addition of new neurons during life in the olfactory bulb is well accepted and has been extensively studied in rodents. This process could allow the animals to adapt to a changing environment. Olfactory neurogenesis begins in the subventricular zone where stem cells proliferate and give rise to young undifferentiated neuroblasts that migrate along the rostral migratory stream to the olfactory bulb (OB). Olfaction is crucial for the expression of sexual behavior in rodents. In female rats, the ability to control the rate of sexual interactions (pacing) has important physiological and behavioral consequences. In the present experiment we evaluated if pacing behavior modifies the rate of new cells that reach the main and accessory olfactory bulb. The BrdU marker was injected before and after different behavioral tests which included: females placed in a mating cage (control), females allowed to pace the sexual interaction, females that mated but were not able to control the rate of the sexual interaction and females exposed to a sexually active male. Subjects were sacrificed fifteen days after the behavioral test. We observed a significant increase in the density of BrdU positive cells in the internal cellular layer of the accessory olfactory bulb when females paced the sexual interaction in comparison to the other 3 groups. No differences in the cell density in the main olfactory bulb were found. These results suggest that pacing behavior promotes an increase in density of the new cells in the accessory olfactory bulb. PMID- 21637744 TI - HIV-1-infected and immune-activated macrophages induce astrocytic differentiation of human cortical neural progenitor cells via the STAT3 pathway. AB - Diminished adult neurogenesis is considered a potential mechanism in the pathogenesis of HIV-1-associated dementia (HAD). In HAD, HIV-1-infected and immune-activated brain mononuclear phagocytes (MP; perivascular macrophages and microglia) drive central nervous system (CNS) inflammation and may alter normal neurogenesis. We previously demonstrated HIV-1-infected and lipopolysaccharide (LPS) activated monocyte-derived macrophages (MDM) inhibit human neural progenitor cell (NPC) neurogenesis, while enhancing astrogliogenesis through the secretion of the inflammatory cytokines such as tumor necrosis factor-alpha (TNF alpha), in vitro and in vivo. Here we further test the hypothesis that HIV-1 infected/activated MDM promote NPC astrogliogenesis via activation of the transcription factor signal transducer and activator of transcription 3 (STAT3), a critical factor for astrogliogenesis. Our results show that LPS-activated MDM conditioned medium (LPS-MCM) and HIV-infected/LPS-activated MDM-conditioned medium (LPS+HIV-MCM) induced Janus kinase 1 (Jak1) and STAT3 activation. Induction of the Jak-STAT3 activation correlated with increased glia fibrillary acidic protein (GFAP) expression, demonstrating an induction of astrogliogenesis. Moreover, STAT3-targeting siRNA (siSTAT3) decreased MCM-induced STAT3 activation and NPC astrogliogenesis. Furthermore, inflammatory cytokines (including IL-6, IL 1beta and TNF-alpha) produced by LPS-activated and/or HIV-1-infected MDM may contribute to MCM-induced STAT3 activation and astrocytic differentiation. These observations were confirmed in severe combined immunodeficient (SCID) mice with HIV-1 encephalitis (HIVE). In HIVE mice, siRNA control (without target sequence, sicon) pre-transfected NPCs injected with HIV-1-infected MDM showed more astrocytic differentiation and less neuronal differentiation of NPCs as compared to NPC injection alone. siSTAT3 abrogated HIV-1-infected MDM-induced astrogliogenesis of injected NPCs. Collectively, these observations demonstrate that HIV-1-infected/activated MDM induces NPC astrogliogenesis through the STAT3 pathway. This study generates important data elucidating the role of brain inflammation in neurogenesis and may provide insight into new therapeutic strategies for HAD. PMID- 21637746 TI - Detection of pathogenic mycobacteria based on functionalized quantum dots coupled with immunomagnetic separation. AB - Mycobacteria have always proven difficult to identify due to their low growth rate and fastidious nature. Therefore molecular biology and more recently nanotechnology, have been exploited from early on for the detection of these pathogens. Here we present the first stage of development of an assay incorporating cadmium selenide quantum dots (QDs) for the detection of mycobacterial surface antigens. The principle of the assay is the separation of bacterial cells using magnetic beads coupled with genus-specific polyclonal antibodies and monoclonal antibodies for heparin-binding hemagglutinin. These complexes are then tagged with anti-mouse biotinylated antibody and finally streptavidin-conjugated QDs which leads to the detection of a fluorescent signal. For the evaluation of performance, the method under study was applied on Mycobacterium bovis BCG and Mycobacterium tuberculosis (positive controls), as well as E. coli and Salmonella spp. that constituted the negative controls. The direct observation of the latter category of samples did not reveal fluorescence as opposed to the mycobacteria mentioned above. The minimum detection limit of the assay was defined to 10(4) bacteria/ml, which could be further decreased by a 1 log when fluorescence was measured with a spectrofluorometer. The method described here can be easily adjusted for any other protein target of either the pathogen or the host, and once fully developed it will be directly applicable on clinical samples. PMID- 21637745 TI - Polymorphisms in stromal genes and susceptibility to serous epithelial ovarian cancer: a report from the Ovarian Cancer Association Consortium. AB - Alterations in stromal tissue components can inhibit or promote epithelial tumorigenesis. Decorin (DCN) and lumican (LUM) show reduced stromal expression in serous epithelial ovarian cancer (sEOC). We hypothesized that common variants in these genes associate with risk. Associations with sEOC among Caucasians were estimated with odds ratios (OR) among 397 cases and 920 controls in two U.S. based studies (discovery set), 436 cases and 1,098 controls in Australia (replication set 1) and a consortium of 15 studies comprising 1,668 cases and 4,249 controls (replication set 2). The discovery set and replication set 1 (833 cases and 2,013 controls) showed statistically homogeneous (P(heterogeneity)>=0.48) decreased risks of sEOC at four variants: DCN rs3138165, rs13312816 and rs516115, and LUM rs17018765 (OR = 0.6 to 0.9; P(trend) = 0.001 to 0.03). Results from replication set 2 were statistically homogeneous (P(heterogeneity)>=0.13) and associated with increased risks at DCN rs3138165 and rs13312816, and LUM rs17018765: all ORs = 1.2; P(trend)<=0.02. The ORs at the four variants were statistically heterogeneous across all 18 studies (P(heterogeneity)<=0.03), which precluded combining. In post-hoc analyses, interactions were observed between each variant and recruitment period (P(interaction)<=0.003), age at diagnosis (P(interaction) = 0.04), and year of diagnosis (P(interaction) = 0.05) in the five studies with available information (1,044 cases, 2,469 controls). We conclude that variants in DCN and LUM are not directly associated with sEOC, and that confirmation of possible effect modification of the variants by non-genetic factors is required. PMID- 21637748 TI - SARS-CoV 9b protein diffuses into nucleus, undergoes active Crm1 mediated nucleocytoplasmic export and triggers apoptosis when retained in the nucleus. AB - BACKGROUND: 9b is an accessory protein of the SARS-CoV. It is a small protein of 98 amino acids and its structure has been solved recently. 9b is known to localize in the extra-nuclear region and has been postulated to possess a nuclear export signal (NES), however the role of NES in 9b functioning is not well understood. PRINCIPAL FINDINGS/METHODOLOGY: In this report, we demonstrate that 9b in the absence of any nuclear localization signal (NLS) enters the nucleus by passive transport. Using various cell cycle inhibitors, we have shown that the nuclear entry of 9b is independent of the cell cycle. Further, we found that 9b interacts with the cellular protein Crm1 and gets exported out of the nucleus using an active NES. We have also revealed that this NES activity influences the half-life of 9b and affects host cell death. We found that an export signal deficient SARS-CoV 9b protein induces apoptosis in transiently transfected cells and showed elevated caspase-3 activity. CONCLUSION/SIGNIFICANCE: Here, we showed that nuclear shuttling of 9b and its interaction with Crm1 are essential for the proper degradation of 9b and blocking the nuclear export of this protein induces apoptosis. This phenomenon may be critical in providing a novel role to the 9b accessory protein of SARS-CoV. PMID- 21637747 TI - Computational identification of transcriptional regulators in human endotoxemia. AB - One of the great challenges in the post-genomic era is to decipher the underlying principles governing the dynamics of biological responses. As modulating gene expression levels is among the key regulatory responses of an organism to changes in its environment, identifying biologically relevant transcriptional regulators and their putative regulatory interactions with target genes is an essential step towards studying the complex dynamics of transcriptional regulation. We present an analysis that integrates various computational and biological aspects to explore the transcriptional regulation of systemic inflammatory responses through a human endotoxemia model. Given a high-dimensional transcriptional profiling dataset from human blood leukocytes, an elementary set of temporal dynamic responses which capture the essence of a pro-inflammatory phase, a counter regulatory response and a dysregulation in leukocyte bioenergetics has been extracted. Upon identification of these expression patterns, fourteen inflammation-specific gene batteries that represent groups of hypothetically 'coregulated' genes are proposed. Subsequently, statistically significant cis regulatory modules (CRMs) are identified and decomposed into a list of critical transcription factors (34) that are validated largely on primary literature. Finally, our analysis further allows for the construction of a dynamic representation of the temporal transcriptional regulatory program across the host, deciphering possible combinatorial interactions among factors under which they might be active. Although much remains to be explored, this study has computationally identified key transcription factors and proposed a putative time dependent transcriptional regulatory program associated with critical transcriptional inflammatory responses. These results provide a solid foundation for future investigations to elucidate the underlying transcriptional regulatory mechanisms under the host inflammatory response. Also, the assumption that coexpressed genes that are functionally relevant are more likely to share some common transcriptional regulatory mechanism seems to be promising, making the proposed framework become essential in unravelling context-specific transcriptional regulatory interactions underlying diverse mammalian biological processes. PMID- 21637749 TI - Persistence increases with diversity and connectance in trophic metacommunities. AB - BACKGROUND: We are interested in understanding if metacommunity dynamics contribute to the persistence of complex spatial food webs subject to colonization-extinction dynamics. We study persistence as a measure of stability of communities within discrete patches, and ask how do species diversity, connectance, and topology influence it in spatially structured food webs. METHODOLOGY/PRINCIPAL FINDINGS: We answer this question first by identifying two general mechanisms linking topology of simple food web modules and persistence at the regional scale. We then assess the robustness of these mechanisms to more complex food webs with simulations based on randomly created and empirical webs found in the literature. We find that linkage proximity to primary producers and food web diversity generate a positive relationship between complexity and persistence in spatial food webs. The comparison between empirical and randomly created food webs reveal that the most important element for food web persistence under spatial colonization-extinction dynamics is the degree distribution: the number of prey species per consumer is more important than their identity. CONCLUSIONS/SIGNIFICANCE: With a simple set of rules governing patch colonization and extinction, we have predicted that diversity and connectance promote persistence at the regional scale. The strength of our approach is that it reconciles the effect of complexity on stability at the local and the regional scale. Even if complex food webs are locally prone to extinction, we have shown their complexity could also promote their persistence through regional dynamics. The framework we presented here offers a novel and simple approach to understand the complexity of spatial food webs. PMID- 21637750 TI - In vivo dynamics of the musculoskeletal system cannot be adequately described using a stiffness-damping-inertia model. AB - BACKGROUND: Visco-elastic properties of the (neuro-)musculoskeletal system play a fundamental role in the control of posture and movement. Often, these properties are described and identified using stiffness-damping-inertia (KBI) models. In such an approach, perturbations are applied to the (neuro-)musculoskeletal system and subsequently KBI-model parameters are optimized to obtain a best fit between simulated and experimentally observed responses. Problems with this approach may arise because a KBI-model neglects critical aspects of the real musculoskeletal system. METHODOLOGY/PRINCIPAL FINDINGS: The purpose of this study was to analyze the relation between the musculoskeletal properties and the stiffness and damping estimated using a KBI-model, to analyze how this relation is affected by the nature of the perturbation and to assess the sensitivity of the estimated stiffness and damping to measurement errors. Our analyses show that the estimated stiffness and damping using KBI-models do not resemble any of the dynamical parameters of the underlying system, not even when the responses are very accurately fitted by the KBI-model. Furthermore, the stiffness and damping depend non-linearly on all the dynamical parameters of the underlying system, influenced by the nature of the perturbation and the time interval over which the KBI-model is optimized. Moreover, our analyses predict a very high sensitivity of estimated parameters to measurement errors. CONCLUSIONS/SIGNIFICANCE: The results of this study suggest that the usage of stiffness-damping-inertia models to investigate the dynamical properties of the musculoskeletal system under control by the CNS should be reconsidered. PMID- 21637752 TI - An in silico modeling approach to understanding the dynamics of sarcoidosis. AB - BACKGROUND: Sarcoidosis is a polygenic disease with diverse phenotypic presentations characterized by an abnormal antigen-mediated Th1 type immune response. At present, progress towards understanding sarcoidosis disease mechanisms and the development of novel treatments is limited by constraints attendant to conducting human research in a rare disease in the absence of relevant animal models. We sought to develop a computational model to enhance our understanding of the pathological mechanisms of and predict potential treatments of sarcoidosis. METHODOLOGY/RESULTS: Based upon the literature, we developed a computational model of known interactions between essential immune cells (antigen presenting macrophages, effector and regulatory T cells) and cytokine mediators (IL-2, TNFalpha, IFNgamma) of granulomatous inflammation during sarcoidosis. The dynamics of these interactions are described by a set of ordinary differential equations. The model predicts bistable switching behavior which is consistent with normal (self-limited) and "sarcoidosis-like" (sustained) activation of the inflammatory components of the system following a single antigen challenge. By perturbing the influence of model components using inhibitors of the cytokine mediators, distinct clinically relevant disease phenotypes were represented. Finally, the model was shown to be useful for pre-clinical testing of therapies based upon molecular targets and dose-effect relationships. CONCLUSIONS/SIGNIFICANCE: Our work illustrates a dynamic computer simulation of granulomatous inflammation scenarios that is useful for the investigation of disease mechanisms and for pre-clinical therapeutic testing. In lieu of relevant in vitro or animal surrogates, our model may provide for the screening of potential therapies for specific sarcoidosis disease phenotypes in advance of expensive clinical trials. PMID- 21637751 TI - Common gene therapy viral vectors do not efficiently penetrate sputum from cystic fibrosis patients. AB - Norwalk virus and human papilloma virus, two viruses that infect humans at mucosal surfaces, have been found capable of rapidly penetrating human mucus secretions. Viral vectors for gene therapy of Cystic Fibrosis (CF) must similarly penetrate purulent lung airway mucus (sputum) to deliver DNA to airway epithelial cells. However, surprisingly little is known about the rates at which gene delivery vehicles penetrate sputum, including viral vectors used in clinical trials for CF gene therapy. We find that sputum spontaneously expectorated by CF patients efficiently traps two viral vectors commonly used in CF gene therapy trials, adenovirus (d~80 nm) and adeno-associated virus (AAV serotype 5; d~20 nm), leading to average effective diffusivities that are ~3,000-fold and 12,000 fold slower than their theoretical speeds in water, respectively. Both viral vectors are slowed by adhesion, as engineered muco-inert nanoparticles with diameters as large as 200 nm penetrate the same sputum samples at rates only ~40 fold reduced compared to in pure water. A limited fraction of AAV exhibit sufficiently fast mobility to penetrate physiologically thick sputum layers, likely because of the lower viscous drag and smaller surface area for adhesion to sputum constituents. Nevertheless, poor penetration of CF sputum is likely a major contributor to the ineffectiveness of viral vector based gene therapy in the lungs of CF patients observed to date. PMID- 21637753 TI - Automated discrimination of brain pathological state attending to complex structural brain network properties: the shiverer mutant mouse case. AB - Neuroimaging classification procedures between normal and pathological subjects are sparse and highly dependent of an expert's clinical criterion. Here, we aimed to investigate whether possible brain structural network differences in the shiverer mouse mutant, a relevant animal model of myelin related diseases, can reflect intrinsic individual brain properties that allow the automatic discrimination between the shiverer and normal subjects. Common structural networks properties between shiverer (C3Fe.SWV Mbp(shi)/Mbp(shi), n = 6) and background control (C3HeB.FeJ, n = 6) mice are estimated and compared by means of three diffusion weighted MRI (DW-MRI) fiber tractography algorithms and a graph framework. Firstly, we found that brain networks of control group are significantly more clustered, modularized, efficient and optimized than those of the shiverer group, which presented significantly increased characteristic path length. These results are in line with previous structural/functional complex brain networks analysis that have revealed topologic differences and brain network randomization associated to specific states of human brain pathology. In addition, by means of network measures spatial representations and discrimination analysis, we show that it is possible to classify with high accuracy to which group each subject belongs, providing also a probability value of being a normal or shiverer subject as an individual anatomical classifier. The obtained correct predictions (e.g., around 91.6-100%) and clear spatial subdivisions between control and shiverer mice, suggest that there might exist specific network subspaces corresponding to specific brain disorders, supporting also the point of view that complex brain network analyses constitutes promising tools in the future creation of interpretable imaging biomarkers. PMID- 21637754 TI - Autistic traits and brain activation during face-to-face conversations in typically developed adults. AB - BACKGROUND: Autism spectrum disorders (ASD) are characterized by impaired social interaction and communication, restricted interests, and repetitive behaviours. The severity of these characteristics is posited to lie on a continuum that extends into the general population. Brain substrates underlying ASD have been investigated through functional neuroimaging studies using functional magnetic resonance imaging (fMRI). However, fMRI has methodological constraints for studying brain mechanisms during social interactions (for example, noise, lying on a gantry during the procedure, etc.). In this study, we investigated whether variations in autism spectrum traits are associated with changes in patterns of brain activation in typically developed adults. We used near-infrared spectroscopy (NIRS), a recently developed functional neuroimaging technique that uses near-infrared light, to monitor brain activation in a natural setting that is suitable for studying brain functions during social interactions. METHODOLOGY: We monitored regional cerebral blood volume changes using a 52-channel NIRS apparatus over the prefrontal cortex (PFC) and superior temporal sulcus (STS), 2 areas implicated in social cognition and the pathology of ASD, in 28 typically developed participants (14 male and 14 female) during face-to-face conversations. This task was designed to resemble a realistic social situation. We examined the correlations of these changes with autistic traits assessed using the Autism Spectrum Quotient (AQ). PRINCIPAL FINDINGS: Both the PFC and STS were significantly activated during face-to-face conversations. AQ scores were negatively correlated with regional cerebral blood volume increases in the left STS during face-to-face conversations, especially in males. CONCLUSIONS: Our results demonstrate successful monitoring of brain function during realistic social interactions by NIRS as well as lesser brain activation in the left STS during face-to-face conversations in typically developed participants with higher levels of autistic traits. PMID- 21637755 TI - Visualisation of Leishmania donovani fluorescent hybrids during early stage development in the sand fly vector. AB - BACKGROUND: The Leishmania protozoan parasites cause devastating human diseases. Leishmania have been considered to replicate clonally, without genetic exchange. However, an accumulation of evidence indicates that there are inter-specific and intra-specific hybrids among natural populations. The first and so far only experimental proof of genetic exchange was obtained in 2009 when double drug resistant Leishmania major hybrids were produced by co-infecting sand flies with two strains carrying different drug resistance markers. However, the location and timing of hybridisation events in sand flies has not been described. METHODOLOGY/PRINCIPAL FINDINGS: Here we have co-infected Phlebotomus perniciosus and Lutzomyia longipalpis with transgenic promastigotes of Leishmania donovani strains carrying hygromycin or neomycin resistance genes and red or green fluorescent markers. Fed females were dissected at different times post bloodmeal (PBM) and examined by fluorescent microscopy or fluorescent activated cell sorting (FACS) followed by confocal microscopy. In mixed infections strains LEM3804 and Gebre-1 reached the cardia and stomodeal valves more rapidly than strains LEM4265 and LV9. Hybrids unequivocally expressing both red and green fluorescence were seen in single flies of both vectors tested, co-infected with LEM4265 and Gebre-1. The hybrids were present as short (procyclic) promastigotes 2 days PBM in the semi-digested blood in the endoperitrophic space. Recovery of a clearly co-expressing hybrid was also achieved by FACS. However, hybrids could not sustain growth in vitro. CONCLUSIONS/SIGNIFICANCE: For the first time, we observed L. donovani hybrids in the sand fly vector, 2 days PBM and described the morphological stages involved. Fluorescence microscopy in combination with FACS allows visualisation and recovery of the progeny of experimental crosses but on this occasion the hybrids were not viable in vitro. Nevertheless, genetic exchange in L. donovani has profound epidemiological significance, because it facilitates the emergence and spread of new phenotypic traits. PMID- 21637756 TI - Tumor angiogenesis and vascular patterning: a mathematical model. AB - Understanding tumor induced angiogenesis is a challenging problem with important consequences for diagnosis and treatment of cancer. Recently, strong evidences suggest the dual role of endothelial cells on the migrating tips and on the proliferating body of blood vessels, in consonance with further events behind lumen formation and vascular patterning. In this paper we present a multi-scale phase-field model that combines the benefits of continuum physics description and the capability of tracking individual cells. The model allows us to discuss the role of the endothelial cells' chemotactic response and proliferation rate as key factors that tailor the neovascular network. Importantly, we also test the predictions of our theoretical model against relevant experimental approaches in mice that displayed distinctive vascular patterns. The model reproduces the in vivo patterns of newly formed vascular networks, providing quantitative and qualitative results for branch density and vessel diameter on the order of the ones measured experimentally in mouse retinas. Our results highlight the ability of mathematical models to suggest relevant hypotheses with respect to the role of different parameters in this process, hence underlining the necessary collaboration between mathematical modeling, in vivo imaging and molecular biology techniques to improve current diagnostic and therapeutic tools. PMID- 21637757 TI - Live tissue imaging shows reef corals elevate pH under their calcifying tissue relative to seawater. AB - The threat posed to coral reefs by changes in seawater pH and carbonate chemistry (ocean acidification) raises the need for a better mechanistic understanding of physiological processes linked to coral calcification. Current models of coral calcification argue that corals elevate extracellular pH under their calcifying tissue relative to seawater to promote skeleton formation, but pH measurements taken from the calcifying tissue of living, intact corals have not been achieved to date. We performed live tissue imaging of the reef coral Stylophora pistillata to determine extracellular pH under the calcifying tissue and intracellular pH in calicoblastic cells. We worked with actively calcifying corals under flowing seawater and show that extracellular pH (pHe) under the calicoblastic epithelium is elevated by ~0.5 and ~0.2 pH units relative to the surrounding seawater in light and dark conditions respectively. By contrast, the intracellular pH (pHi) of the calicoblastic epithelium remains stable in the light and dark. Estimates of aragonite saturation states derived from our data indicate the elevation in subcalicoblastic pHe favour calcification and may thus be a critical step in the calcification process. However, the observed close association of the calicoblastic epithelium with the underlying crystals suggests that the calicoblastic cells influence the growth of the coral skeleton by other processes in addition to pHe modification. The procedure used in the current study provides a novel, tangible approach for future investigations into these processes and the impact of environmental change on the cellular mechanisms underpinning coral calcification. PMID- 21637758 TI - Combining computational prediction of cis-regulatory elements with a new enhancer assay to efficiently label neuronal structures in the medaka fish. AB - The developing vertebrate nervous system contains a remarkable array of neural cells organized into complex, evolutionarily conserved structures. The labeling of living cells in these structures is key for the understanding of brain development and function, yet the generation of stable lines expressing reporter genes in specific spatio-temporal patterns remains a limiting step. In this study we present a fast and reliable pipeline to efficiently generate a set of stable lines expressing a reporter gene in multiple neuronal structures in the developing nervous system in medaka. The pipeline combines both the accurate computational genome-wide prediction of neuronal specific cis-regulatory modules (CRMs) and a newly developed experimental setup to rapidly obtain transgenic lines in a cost-effective and highly reproducible manner. 95% of the CRMs tested in our experimental setup show enhancer activity in various and numerous neuronal structures belonging to all major brain subdivisions. This pipeline represents a significant step towards the dissection of embryonic neuronal development in vertebrates. PMID- 21637759 TI - Time changes with the embodiment of another's body posture. AB - The aim of the present study was to investigate whether the perception of presentation durations of pictures of different body postures was distorted as function of the embodied movement that originally produced these postures. Participants were presented with two pictures, one with a low-arousal body posture judged to require no movement and the other with a high-arousal body posture judged to require considerable movement. In a temporal bisection task with two ranges of standard durations (0.4/1.6 s and 2/8 s), the participants had to judge whether the presentation duration of each of the pictures was more similar to the short or to the long standard duration. The results showed that the duration was judged longer for the posture requiring more movement than for the posture requiring less movement. However the magnitude of this overestimation was relatively greater for the range of short durations than for that of longer durations. Further analyses suggest that this lengthening effect was mediated by an arousal effect of limited duration on the speed of the internal clock system. PMID- 21637760 TI - Monitoring of in vivo function of superparamagnetic iron oxide labelled murine dendritic cells during anti-tumour vaccination. AB - Dendritic cells (DCs) generated in vitro to present tumour antigens have been injected in cancer patients to boost in vivo anti-tumour immune responses. This approach to cancer immunotherapy has had limited success. For anti-tumour therapy, delivery and subsequent migration of DCs to lymph nodes leading to effective stimulation of effector T cells is thought to be essential. The ability to non-invasively monitor the fate of adoptively transferred DCs in vivo using magnetic resonance imaging (MRI) is an important clinical tool to correlate their in vivo behavior with response to treatment. Previous reports of superparamagnetic iron oxides (SPIOs) labelling of different cell types, including DCs, have indicated varying detrimental effects on cell viability, migration, differentiation and immune function. Here we describe an optimised labelling procedure using a short incubation time and low concentration of clinically used SPIO Endorem to successfully track murine DC migration in vivo using MRI in a mouse tumour model. First, intracellular labelling of bone marrow derived DCs was monitored in vitro using electron microscopy and MRI relaxometry. Second, the in vitro characterisation of SPIO labelled DCs demonstrated that viability, phenotype and functions were comparable to unlabelled DCs. Third, ex vivo SPIO labelled DCs, when injected subcutaneously, allowed for the longitudinal monitoring by MR imaging of their migration in vivo. Fourth, the SPIO DCs induced the proliferation of adoptively transferred CD4(+) T cells but, most importantly, they primed cytotoxic CD8(+) T cell responses to protect against a B16-Ova tumour challenge. Finally, using anatomical information from the MR images, the immigration of DCs was confirmed by the increase in lymph node size post-DC injection. These results demonstrate that the SPIO labelling protocol developed in this study is not detrimental for DC function in vitro and in vivo has potential clinical application in monitoring therapeutic DCs in patients with cancer. PMID- 21637761 TI - Thyroid hormone T3 counteracts STZ induced diabetes in mouse. AB - This study intended to demonstrate that the thyroid hormone T3 counteracts the onset of a Streptozotocin (STZ) induced diabetes in wild type mice. To test our hypothesis diabetes has been induced in Balb/c male mice by multiple low dose Streptozotocin injection; and a group of mice was contemporaneously injected with T3. After 48 h mice were tested for glucose tolerance test, insulin serum levels and then sacrificed. Whole pancreata were utilized for morphological and biochemical analyses, while protein extracts and RNA were utilized for expression analyses of specific molecules. The results showed that islets from T3 treated mice were comparable to age- and sex-matched control, untreated mice in number, shape, dimension, consistency, ultrastructure, insulin and glucagon levels, Tunel positivity and caspases activation, while all the cited parameters and molecules were altered by STZ alone. The T3-induced pro survival effect was associated with a strong increase in phosphorylated Akt. Moreover, T3 administration prevented the STZ-dependent alterations in glucose blood level, both during fasting and after glucose challenge, as well as in insulin serum level. In conclusion we demonstrated that T3 could act as a protective factor against STZ induced diabetes. PMID- 21637762 TI - Adiponectin gene polymorphism is selectively associated with the concomitant presence of metabolic syndrome and essential hypertension. AB - OBJECTIVE: Cardiovascular risk increases with the presence of both metabolic syndrome (MetS) and hypertension (HTN). Although the adiponectin (ADIPOQ) gene has been reported to be involved in MetS, its association with HTN remained undetermined. This study aimed to investigate the association of ADIPOQ gene with the phenotypes of HTN and MetS. METHODS: A total of 962 participants from 302 families from the Taiwan young-onset hypertension genetic study were enrolled. Plasma adiponectin were measured, and association analysis was conducted by using GEE regression-based method. Another study, of 1448 unrelated participants, was conducted to replicate the association between ADIPOQ gene and variable phenotypes of MetS with or without HTN. RESULTS: Among 962 subjects from family samples, the lowest plasma adiponectin value was observed in MetS with HTN component (9.3+/-0.47 ug/ml) compared with hypertensives (13.4+/-0.74 ug /ml) or MetS without HTN (11.9+/-0.60 ug/ml, P<0.05). The SNP rs1501299 (G276T) in ADIPOQ gene was found associated with the presence of HTN in MetS (odds ratio for GG+GT vs. TT = 2.46; 95% CI: 1.14-5.3, p = 0.02), but not rs2241766 (T45G). No association of ADIPOQ gene with HTN alone or MetS without HTN was observed. The significant association of the SNP rs1501299 (G276T) with the phenotype of presence of HTN in MetS was confirmed (odds ratio for GG+GT vs. TT = 2.15; 95% CI: 1.1-4.3) in the replication study. CONCLUSIONS: ADIPOQ genetic variants were selectively and specifically associated with the concomitant presence of MetS and HTN, suggesting potential genetic linkage between MetS and HTN. PMID- 21637763 TI - Comparison of human memory CD8 T cell responses to adenoviral early and late proteins in peripheral blood and lymphoid tissue. AB - Treatment of invasive adenovirus (Ad) disease in hematopoietic stem cell transplant (SCT) recipients with capsid protein hexon-specific donor T cells is under investigation. We propose that cytotoxic T cells (CTLs) targeted to the late protein hexon may be inefficient in vivo because the early Ad protein E3-19K downregulates HLA class I antigens in infected cells. In this study, CD8+ T cells targeted to highly conserved HLA A2-restricted epitopes from the early regulatory protein DNA polymerase (P-977) and late protein hexon (H-892) were compared in peripheral blood (PB) and tonsils of naturally infected adults. In tonsils, epitope-specific pentamers detected a significantly higher frequency of P 977+CD8+ T cells compared to H-892+CD8+ T cells; this trend was reversed in PB. Tonsil epitope-specific CD8+ T cells expressed IFN-gamma and IL-2 but not perforin or TNF-alpha, whereas PB T cells were positive for IFN-gamma, TNF-alpha, and perforin. Tonsil epitope-specific T cells expressed lymphoid homing marker CCR7 and exhibited lower levels of the activation marker CD25 but higher proliferative potential than PB T cells. Finally, in parallel with the kinetics of mRNA expression, P-977-specific CTLs lysed targets as early as 8 hrs post infection. In contrast, H-892-specific CTLs did not kill unless infected fibroblasts were pretreated with IFN-gamma to up regulate HLA class I antigens, and cytotoxicity was delayed until 16-24 hours. These data show that, in contrast to hexon CTLs, central memory type DNA polymerase CTLs dominate the lymphoid compartment and kill fibroblasts earlier after infection without requiring exogenous IFN-gamma. Thus, use of CTLs targeted to both early and late Ad proteins may improve the efficacy of immunotherapy for life-threatening Ad disease in SCT recipients. PMID- 21637764 TI - Genetic and environmental influences on infant growth: prospective analysis of the Gemini twin birth cohort. AB - OBJECTIVE: Infancy is a critical period during which rapid growth potentially programs future disease risk. Identifying the modifiable determinants of growth is therefore important. To capture the complexity of infant growth, we modeled growth trajectories from birth to six months in order to compare the genetic and environmental influences on growth trajectory parameters with single time-point measures at birth, three and six months of age. METHODS: Data were from Gemini, a population sample of 2402 UK families with twins. An average 10 weight measurements per child made by health professionals were available over the first six months. Weights at birth, three and six months were identified. Longitudinal growth trajectories were modeled using SITAR utilizing all available weight measures for each child. SITAR generates three parameters: size (characterizing mean weight throughout infancy), tempo (indicating age at peak weight velocity (PWV)), and velocity (reflecting the size of PWV). Genetic and environmental influences were estimated using quantitative genetic analysis. RESULTS: In line with previous studies, heritability of weight at birth and three months was low (38%), but it was higher at six months (62%). Heritability of the growth trajectory parameters was high for size (69%) and velocity (57%), but low (35%) for tempo. Common environmental influences predominated for tempo (42%). CONCLUSION: Modeled growth parameters using SITAR indicated that size and velocity were primarily under genetic influence but tempo was predominantly environmentally determined. These results emphasize the importance of identifying specific modifiable environmental determinants of the timing of peak infant growth. PMID- 21637765 TI - Epithelial membrane protein-2 promotes endometrial tumor formation through activation of FAK and Src. AB - Endometrial cancer is the most common gynecologic malignancy diagnosed among women in developed countries. One recent biomarker strongly associated with disease progression and survival is epithelial membrane protein-2 (EMP2), a tetraspan protein known to associate with and modify surface expression of certain integrin isoforms. In this study, we show using a xenograft model system that EMP2 expression is necessary for efficient endometrial tumor formation, and we have started to characterize the mechanism by which EMP2 contributes to this malignant phenotype. In endometrial cancer cells, the focal adhesion kinase (FAK)/Src pathway appears to regulate migration as measured through wound healing assays. Manipulation of EMP2 levels in endometrial cancer cells regulates the phosphorylation of FAK and Src, and promotes their distribution into lipid raft domains. Notably, cells with low levels of EMP2 fail to migrate and poorly form tumors in vivo. These findings reveal the pivotal role of EMP2 in endometrial cancer carcinogenesis, and suggest that the association of elevated EMP2 levels with endometrial cancer prognosis may be causally linked to its effect on integrin-mediated signaling. PMID- 21637766 TI - Progression of biopsy-measured liver fibrosis in untreated patients with hepatitis C infection: non-Markov multistate model analysis. AB - BACKGROUND: Fibrosis stages from liver biopsies reflect liver damage from hepatitis C infection, but analysis is challenging due to their ordered but non numeric nature, infrequent measurement, misclassification, and unknown infection times. METHODS: We used a non-Markov multistate model, accounting for misclassification, with multiple imputation of unknown infection times, applied to 1062 participants of whom 159 had multiple biopsies. Odds ratios (OR) quantified the estimated effects of covariates on progression risk at any given time. RESULTS: Models estimated that progression risk decreased the more time participants had already spent in the current stage, African American race was protective (OR 0.75, 95% confidence interval 0.60 to 0.95, p = 0.018), and older current age increased risk (OR 1.33 per decade, 95% confidence interval 1.15 to 1.54, p = 0.0002). When controlled for current age, older age at infection did not appear to increase risk (OR 0.92 per decade, 95% confidence interval 0.47 to 1.79, p = 0.80). There was a suggestion that co-infection with human immunodeficiency virus increased risk of progression in the era of highly active antiretroviral treatment beginning in 1996 (OR 2.1, 95% confidence interval 0.97 to 4.4, p = 0.059). Other examined risk factors may influence progression risk, but evidence for or against this was weak due to wide confidence intervals. The main results were essentially unchanged using different assumed misclassification rates or imputation of age of infection. DISCUSSION: The analysis avoided problems inherent in simpler methods, supported the previously suspected protective effect of African American race, and suggested that current age rather than age of infection increases risk. Decreasing risk of progression with longer time already spent in a stage was also previously found for post-transplant progression. This could reflect varying disease activity, with recent progression indicating active disease and high risk, while longer time already spent in a stage indicates quiescent disease and low risk. PMID- 21637767 TI - Effect of the topology and delayed interactions in neuronal networks synchronization. AB - As important as the intrinsic properties of an individual nervous cell stands the network of neurons in which it is embedded and by virtue of which it acquires great part of its responsiveness and functionality. In this study we have explored how the topological properties and conduction delays of several classes of neural networks affect the capacity of their constituent cells to establish well-defined temporal relations among firing of their action potentials. This ability of a population of neurons to produce and maintain a millisecond-precise coordinated firing (either evoked by external stimuli or internally generated) is central to neural codes exploiting precise spike timing for the representation and communication of information. Our results, based on extensive simulations of conductance-based type of neurons in an oscillatory regime, indicate that only certain topologies of networks allow for a coordinated firing at a local and long range scale simultaneously. Besides network architecture, axonal conduction delays are also observed to be another important factor in the generation of coherent spiking. We report that such communication latencies not only set the phase difference between the oscillatory activity of remote neural populations but determine whether the interconnected cells can set in any coherent firing at all. In this context, we have also investigated how the balance between the network synchronizing effects and the dispersive drift caused by inhomogeneities in natural firing frequencies across neurons is resolved. Finally, we show that the observed roles of conduction delays and frequency dispersion are not particular to canonical networks but experimentally measured anatomical networks such as the macaque cortical network can display the same type of behavior. PMID- 21637768 TI - Polyhydroxy fullerenes (fullerols or fullerenols): beneficial effects on growth and lifespan in diverse biological models. AB - Recent toxicological studies on carbon nanomaterials, including fullerenes, have led to concerns about their safety. Functionalized fullerenes, such as polyhydroxy fullerenes (PHF, fullerols, or fullerenols), have attracted particular attention due to their water solubility and toxicity. Here, we report surprisingly beneficial and/or specific effects of PHF on model organisms representing four kingdoms, including the green algae Pseudokirchneriella subcapitata, the plant Arabidopsis thaliana, the fungus Aspergillus niger, and the invertebrate Ceriodaphnia dubia. The results showed that PHF had no acute or chronic negative effects on the freshwater organisms. Conversely, PHF could surprisingly increase the algal culture density over controls at higher concentrations (i.e., 72% increase by 1 and 5 mg/L of PHF) and extend the lifespan and stimulate the reproduction of Daphnia (e.g. about 38% by 20 mg/L of PHF). We also show that at certain PHF concentrations fungal growth can be enhanced and Arabidopsis thaliana seedlings exhibit longer hypocotyls, while other complex physiological processes remain unaffected. These findings may open new research fields in the potential applications of PHF, e.g., in biofuel production and aquaculture. These results will form the basis of further research into the mechanisms of growth stimulation and life extension by PHF. PMID- 21637769 TI - A multiwell platform for studying stiffness-dependent cell biology. AB - Adherent cells are typically cultured on rigid substrates that are orders of magnitude stiffer than their tissue of origin. Here, we describe a method to rapidly fabricate 96 and 384 well platforms for routine screening of cells in tissue-relevant stiffness contexts. Briefly, polyacrylamide (PA) hydrogels are cast in glass-bottom plates, functionalized with collagen, and sterilized for cell culture. The Young's modulus of each substrate can be specified from 0.3 to 55 kPa, with collagen surface density held constant over the stiffness range. Using automated fluorescence microscopy, we captured the morphological variations of 7 cell types cultured across a physiological range of stiffness within a 384 well plate. We performed assays of cell number, proliferation, and apoptosis in 96 wells and resolved distinct profiles of cell growth as a function of stiffness among primary and immortalized cell lines. We found that the stiffness-dependent growth of normal human lung fibroblasts is largely invariant with collagen density, and that differences in their accumulation are amplified by increasing serum concentration. Further, we performed a screen of 18 bioactive small molecules and identified compounds with enhanced or reduced effects on soft versus rigid substrates, including blebbistatin, which abolished the suppression of lung fibroblast growth at 1 kPa. The ability to deploy PA gels in multiwell plates for high throughput analysis of cells in tissue-relevant environments opens new opportunities for the discovery of cellular responses that operate in specific stiffness regimes. PMID- 21637770 TI - DRD4-exonIII-VNTR moderates the effect of childhood adversities on emotional resilience in young-adults. AB - Most individuals successfully maintain psychological well-being even when exposed to trauma or adversity. Emotional resilience or the ability to thrive in the face of adversity is determined by complex interactions between genetic makeup, previous exposure to stress, personality, coping style, availability of social support, etc. Recent studies have demonstrated that childhood trauma diminishes resilience in adults and affects mental health. The Dopamine receptor D4 (DRD4) exon III variable number tandem repeat (VNTR) polymorphism was reported to moderate the impact of adverse childhood environment on behaviour, mood and other health-related outcomes. In this study we investigated whether DRD4-exIII-VNTR genotype moderates the effect of childhood adversities (CA) on resilience. In a representative population sample (n = 1148) aged 30-34 years, we observed an interactive effect of DRD4 genotype and CA (beta = 0.132; p = 0.003) on resilience despite no main effect of the genotype when effects of age, gender and education were controlled for. The 7-repeat allele appears to protect against the adverse effect of CA since the decline in resilience associated with increased adversity was evident only in individuals without the 7-repeat allele. Resilience was also significantly associated with approach-/avoidance-related personality measures (behavioural inhibition/activation system; BIS/BAS) measures and an interactive effect of DRD4-exIII-VNTR genotype and CA on BAS was observed. Hence it is possible that approach-related personality traits could be mediating the effect of the DRD4 gene and childhood environment interaction on resilience such that when stressors are present, the 7-repeat allele influences the development of personality in a way that provides protection against adverse outcomes. PMID- 21637771 TI - Effects of common polymorphisms rs11614913 in miR-196a2 and rs2910164 in miR-146a on cancer susceptibility: a meta-analysis. AB - BACKGROUND: MicroRNAs regulate gene expression at the post-transcriptional level and involved in diverse biological and pathological processes, including tumorigenesis. Rs11614913 in miR-196a2 and rs2910164 in miR-146a are shown to associate with increased/decreased cancer risk. We performed a meta-analysis to systematically summarize the possible association. METHODOLOGY/PRINCIPAL FINDINGS: We assessed published studies of the association between these microRNA polymorphisms and cancer risk from eleven studies with 16,771 subjects for miR 196a2 and from ten studies with 15,126 subjects for miR-146a. As for rs11614913, the contrast of homozygote (TT vs CC: OR = 0.92, 95% CI = 0.85-0.99, P(heterogeneity) = 0.45), allele (T vs C: OR = 0.96, 95% CI = 0.92-0.99, P(heterogeneity) = 0.61) and recessive model (OR = 0.90, 95% CI = 0.84-0.97, P(heterogeneity) = 0.50) produced statistically association. Subgroup analysis by ethnicity, statistically significantly decreased cancer risks were found among Asians for allele contrast (OR = 0.95, 95% CI = 0.90-0.99, P(heterogeneity) = 0.74) and the recessive genetic model (OR = 0.90, 95% CI = 0.82-0.98, P(heterogeneity) = 0.85). According to subgroup analysis by tumor types, the protective effect of C/T polymorphism was only found in breast cancer under allele contrast (T vs C: OR = 0.94, 95% CI = 0.88-0.99, P(heterogeneity) = 0.26). For rs2910164, no significant associations were found among overall analysis model with relatively large heterogeneity. Through the stratified analysis, heterogeneity decreased significantly. In the subgroup analyses by cancer types, the C allele of rs2910164 was associated with protection from digestive cancer in allele contrast (C vs G: OR = 0.86, 95% CI = 0.77-0.96, P(heterogeneity) = 0.51). CONCLUSIONS/SIGNIFICANCE: Our meta-analysis suggests that the rs11614913 most likely contributes to decreased susceptibility to cancer, especially in Asians and breast cancer. Besides, the C allele of the rs2910164 might be associated with a protection from digestive cancer. PMID- 21637772 TI - ALTERED MERISTEM PROGRAM 1 is involved in development of seed dormancy in Arabidopsis. AB - Mutants in the rice PLASTOCHRON 3 and maize VIVIPAROUS 8 genes have been shown to have reduced dormancy and ABA levels. In this study we used several mutants in the orthologous gene ALTERED MERISTEM PROGRAM 1 (AMP1) to determine its role in seed dormancy in Arabidopsis. Here we report that there are accession-specific effects of mutations in AMP1. In one accession, amp1 mutants produce seeds with higher dormancy, while those in two other accessions produce seeds of lower dormancy. These accession-specific effects of mutating AMP1 were shown to extend to ABA levels. We assayed global gene transcription differences in seeds of wild type and mutant from two accessions demonstrating opposing phenotypes. The transcript changes observed indicate that the amp1 mutation shifts the seed transcriptome from a dormant into an after-ripened state. Specific changes in gene expression in the mutants give insight into the direct and indirect effects that may be contributing to the opposing dormancy phenotypes observed, and reveal a role for AMP1 in the acquisition and/or maintenance of seed dormancy in Arabidopsis. PMID- 21637773 TI - MDA5 and TLR3 initiate pro-inflammatory signaling pathways leading to rhinovirus induced airways inflammation and hyperresponsiveness. AB - Rhinovirus (RV), a single-stranded RNA picornavirus, is the most frequent cause of asthma exacerbations. We previously demonstrated in human bronchial epithelial cells that melanoma differentiation-associated gene (MDA)-5 and the adaptor protein for Toll-like receptor (TLR)-3 are each required for maximal RV1B-induced interferon (IFN) responses. However, in vivo, the overall airway response to viral infection likely represents a coordinated response integrating both antiviral and pro-inflammatory pathways. We examined the airway responses of MDA5 and TLR3-deficient mice to infection with RV1B, a minor group virus which replicates in mouse lungs. MDA5 null mice showed a delayed type I IFN and attenuated type III IFN response to RV1B infection, leading to a transient increase in viral titer. TLR3 null mice showed normal IFN responses and unchanged viral titers. Further, RV-infected MDA5 and TLR3 null mice showed reduced lung inflammatory responses and reduced airways responsiveness. Finally, RV-infected MDA5 null mice with allergic airways disease showed lower viral titers despite deficient IFN responses, and allergic MDA5 and TLR3 null mice each showed decreased RV-induced airway inflammatory and contractile responses. These results suggest that, in the context of RV infection, binding of viral dsRNA to MDA5 and TLR3 initiates pro-inflammatory signaling pathways leading to airways inflammation and hyperresponsiveness. PMID- 21637774 TI - Arachnids secrete a fluid over their adhesive pads. AB - BACKGROUND: Many arachnids possess adhesive pads on their feet that help them climb smooth surfaces and capture prey. Spider and gecko adhesives have converged on a branched, hairy structure, which theoretically allows them to adhere solely by dry (solid-solid) intermolecular interactions. Indeed, the consensus in the literature is that spiders and their smooth-padded relatives, the solifugids, adhere without the aid of a secretion. METHODOLOGY AND PRINCIPAL FINDINGS: We investigated the adhesive contact zone of living spiders, solifugids and mites using interference reflection microscopy, which allows the detection of thin liquid films. Like insects, all the arachnids we studied left behind hydrophobic fluid footprints on glass (mean refractive index: 1.48-1.50; contact angle: 3.7 11.2 degrees ). Fluid was not always secreted continuously, suggesting that pads can function in both wet and dry modes. We measured the attachment forces of single adhesive setae from tarantulas (Grammostola rosea) by attaching them to a bending beam with a known spring constant and filming the resulting deflection. Individual spider setae showed a lower static friction at rest (26%+/-2.8 SE of the peak friction) than single gecko setae (Thecadactylus rapicauda; 96%+/-1.7 SE). This may be explained by the fact that spider setae continued to release fluid after isolation from the animal, lubricating the contact zone. SIGNIFICANCE: This finding implies that tarsal secretions occur within all major groups of terrestrial arthropods with adhesive pads. The presence of liquid in an adhesive contact zone has important consequences for attachment performance, improving adhesion to rough surfaces and introducing rate-dependent effects. Our results leave geckos and anoles as the only known representatives of truly dry adhesive pads in nature. Engineers seeking biological inspiration for synthetic adhesives should consider whether model species with fluid secretions are appropriate to their design goals. PMID- 21637775 TI - Crystal structure of a novel esterase Rv0045c from Mycobacterium tuberculosis. AB - There are at least 250 enzymes in Mycobacterium tuberculosis (M. tuberculosis) involved in lipid metabolism. Some of the enzymes are required for bacterial survival and full virulence. The esterase Rv0045c shares little amino acid sequence similarity with other members of the esterase/lipase family. Here, we report the 3D structure of Rv0045c. Our studies demonstrated that Rv0045c is a novel member of alpha/beta hydrolase fold family. The structure of esterase Rv0045c contains two distinct domains: the alpha/beta fold domain and the cap domain. The active site of esterase Rv0045c is highly conserved and comprised of two residues: Ser154 and His309. We proposed that Rv0045c probably employs two kinds of enzymatic mechanisms when hydrolyzing C-O ester bonds within substrates. The structure provides insight into the hydrolysis mechanism of the C-O ester bond, and will be helpful in understanding the ester/lipid metabolism in M. tuberculosis. PMID- 21637777 TI - Comparison of two porcine-derived materials for repairing abdominal wall defects in rats. AB - OBJECTIVE: The purpose of this study was to compare the mechanical properties, host responses and incorporation of porcine small intestine submucosa (PSIS) and porcine acellular dermal matrix (PADM) in a rat model of abdominal wall defect repair. MATERIALS AND METHODS: Prior to implantation, PSIS and PADM were prepared and evaluated in terms of structure and mechanical properties. Full-thickness abdominal wall defects were created in 50 Sprague-Dawley rats, and were repaired using either PSIS or PADM. Rats were sacrificed 1, 2, 4, 8 and 12 weeks post repair and examined for herniation, infection, adhesions, contraction, and changes in the thickness and strength of the tissues incorporated at the defect sites. Histopathology and immunohistochemistry were performed to analyze inflammatory responses, collagen deposition and vascularization. RESULTS: PADM showed more dense collagen deposition and stronger mechanical properties than PSIS prior to implantation (P<0.01). However, the mechanical properties observed after integration with the surrounding native tissues was similar for PADM and PSIS. Both PADM and PSIS showed significant contraction by week 12. However, PADM tissue induced less adhesion and increased in thickness more slowly, and showed less infiltration by foreign giant cells, polymorphonuclear cells, and mononuclear cells. Improved remodeling of host tissue was observed after PSIS implantation, which was apparent from the orientation of bands of fibrous connective tissue, intermixed with newly formed blood vessels by Week 12. CONCLUSION: PSIS showed weaker mechanical properties prior to implantation. However, after implantation PSIS induced more pronounced host responses and showed better incorporation into host tissues than PADM. PMID- 21637776 TI - Criterion and construct validity of the CogState Schizophrenia Battery in Japanese patients with schizophrenia. AB - BACKGROUND: The CogState Schizophrenia Battery (CSB), a computerized cognitive battery, covers all the same cognitive domains as the Measurement and Treatment Research to Improve Cognition in Schizophrenia (MATRICS) Consensus Cognitive Battery but is briefer to conduct. The aim of the present study was to evaluate the criterion and construct validity of the Japanese language version of the CSB (CSB-J) in Japanese patients with schizophrenia. METHODOLOGY/PRINCIPAL FINDINGS: Forty Japanese patients with schizophrenia and 40 Japanese healthy controls with matching age, gender, and premorbid intelligence quotient were enrolled. The CSB J and the Brief Assessment of Cognition in Schizophrenia, Japanese-language version (BACS-J) were performed once. The structure of the CSB-J was also evaluated by a factor analysis. Similar to the BACS-J, the CSB-J was sensitive to cognitive impairment in Japanese patients with schizophrenia. Furthermore, there was a significant positive correlation between the CSB-J composite score and the BACS-J composite score. A factor analysis showed a three-factor model consisting of memory, speed, and social cognition factors. CONCLUSIONS/SIGNIFICANCE: This study suggests that the CSB-J is a useful and rapid automatically administered computerized battery for assessing broad cognitive domains in Japanese patients with schizophrenia. PMID- 21637778 TI - STAT is an essential activator of the zygotic genome in the early Drosophila embryo. AB - In many organisms, transcription of the zygotic genome begins during the maternal to-zygotic transition (MZT), which is characterized by a dramatic increase in global transcriptional activities and coincides with embryonic stem cell differentiation. In Drosophila, it has been shown that maternal morphogen gradients and ubiquitously distributed general transcription factors may cooperate to upregulate zygotic genes that are essential for pattern formation in the early embryo. Here, we show that Drosophila STAT (STAT92E) functions as a general transcription factor that, together with the transcription factor Zelda, induces transcription of a large number of early-transcribed zygotic genes during the MZT. STAT92E is present in the early embryo as a maternal product and is active around the MZT. DNA-binding motifs for STAT and Zelda are highly enriched in promoters of early zygotic genes but not in housekeeping genes. Loss of Stat92E in the early embryo, similarly to loss of zelda, preferentially down regulates early zygotic genes important for pattern formation. We further show that STAT92E and Zelda synergistically regulate transcription. We conclude that STAT92E, in conjunction with Zelda, plays an important role in transcription of the zygotic genome at the onset of embryonic development. PMID- 21637779 TI - Characterizing genetic risk at known prostate cancer susceptibility loci in African Americans. AB - GWAS of prostate cancer have been remarkably successful in revealing common genetic variants and novel biological pathways that are linked with its etiology. A more complete understanding of inherited susceptibility to prostate cancer in the general population will come from continuing such discovery efforts and from testing known risk alleles in diverse racial and ethnic groups. In this large study of prostate cancer in African American men (3,425 prostate cancer cases and 3,290 controls), we tested 49 risk variants located in 28 genomic regions identified through GWAS in men of European and Asian descent, and we replicated associations (at p<=0.05) with roughly half of these markers. Through fine mapping, we identified nearby markers in many regions that better define associations in African Americans. At 8q24, we found 9 variants (p<=6*10(-4)) that best capture risk of prostate cancer in African Americans, many of which are more common in men of African than European descent. The markers found to be associated with risk at each locus improved risk modeling in African Americans (per allele OR = 1.17) over the alleles reported in the original GWAS (OR = 1.08). In summary, in this detailed analysis of the prostate cancer risk loci reported from GWAS, we have validated and improved upon markers of risk in some regions that better define the association with prostate cancer in African Americans. Our findings with variants at 8q24 also reinforce the importance of this region as a major risk locus for prostate cancer in men of African ancestry. PMID- 21637780 TI - DNA methylation dynamics in human induced pluripotent stem cells over time. AB - Epigenetic reprogramming is a critical event in the generation of induced pluripotent stem cells (iPSCs). Here, we determined the DNA methylation profiles of 22 human iPSC lines derived from five different cell types (human endometrium, placental artery endothelium, amnion, fetal lung fibroblast, and menstrual blood cell) and five human embryonic stem cell (ESC) lines, and we followed the aberrant methylation sites in iPSCs for up to 42 weeks. The iPSCs exhibited distinct epigenetic differences from ESCs, which were caused by aberrant methylation at early passages. Multiple appearances and then disappearances of random aberrant methylation were detected throughout iPSC reprogramming. Continuous passaging of the iPSCs diminished the differences between iPSCs and ESCs, implying that iPSCs lose the characteristics inherited from the parent cells and adapt to very closely resemble ESCs over time. Human iPSCs were gradually reprogrammed through the "convergence" of aberrant hyper-methylation events that continuously appeared in a de novo manner. This iPS reprogramming consisted of stochastic de novo methylation and selection/fixation of methylation in an environment suitable for ESCs. Taken together, random methylation and convergence are driving forces for long-term reprogramming of iPSCs to ESCs. PMID- 21637781 TI - SHINE transcription factors act redundantly to pattern the archetypal surface of Arabidopsis flower organs. AB - Floral organs display tremendous variation in their exterior that is essential for organogenesis and the interaction with the environment. This diversity in surface characteristics is largely dependent on the composition and structure of their coating cuticular layer. To date, mechanisms of flower organ initiation and identity have been studied extensively, while little is known regarding the regulation of flower organs surface formation, cuticle composition, and its developmental significance. Using a synthetic microRNA approach to simultaneously silence the three SHINE (SHN) clade members, we revealed that these transcription factors act redundantly to shape the surface and morphology of Arabidopsis flowers. It appears that SHNs regulate floral organs' epidermal cell elongation and decoration with nanoridges, particularly in petals. Reduced activity of SHN transcription factors results in floral organs' fusion and earlier abscission that is accompanied by a decrease in cutin load and modified cell wall properties. SHN transcription factors possess target genes within four cutin- and suberin-associated protein families including, CYP86A cytochrome P450s, fatty acyl-CoA reductases, GSDL-motif lipases, and BODYGUARD1-like proteins. The results suggest that alongside controlling cuticular lipids metabolism, SHNs act to modify the epidermis cell wall through altering pectin metabolism and structural proteins. We also provide evidence that surface formation in petals and other floral organs during their growth and elongation or in abscission and dehiscence through SHNs is partially mediated by gibberellin and the DELLA signaling cascade. This study therefore demonstrates the need for a defined composition and structure of the cuticle and cell wall in order to form the archetypal features of floral organs surfaces and control their cell-to-cell separation processes. Furthermore, it will promote future investigation into the relation between the regulation of organ surface patterning and the broader control of flower development and biological functions. PMID- 21637782 TI - Vascular wall-resident CD44+ multipotent stem cells give rise to pericytes and smooth muscle cells and contribute to new vessel maturation. AB - Here, we identify CD44(+)CD90(+)CD73(+)CD34(-)CD45(-) cells within the adult human arterial adventitia with properties of multipotency which were named vascular wall-resident multipotent stem cells (VW-MPSCs). VW-MPSCs exhibit typical mesenchymal stem cell characteristics including cell surface markers in immunostaining and flow cytometric analyses, and differentiation into adipocytes, chondrocytes and osteocytes under culture conditions. Particularly, TGFbeta1 stimulation up-regulates smooth muscle cell markers in VW-MPSCs. Using fluorescent cell labelling and co-localisation studies we show that VW-MPSCs differentiate to pericytes/smooth muscle cells which cover the wall of newly formed endothelial capillary-like structures in vitro. Co-implantation of EGFP labelled VW-MPSCs and human umbilical vein endothelial cells into SCID mice subcutaneously via Matrigel results in new vessels formation which were covered by pericyte- or smooth muscle-like cells generated from implanted VW-MPSCs. Our results suggest that VW-MPSCs are of relevance for vascular morphogenesis, repair and self-renewal of vascular wall cells and for local capacity of neovascularization in disease processes. PMID- 21637783 TI - Microsatellite instability in pediatric high grade glioma is associated with genomic profile and differential target gene inactivation. AB - High grade gliomas (HGG) are one of the leading causes of cancer-related deaths in children, and there is increasing evidence that pediatric HGG may harbor distinct molecular characteristics compared to adult tumors. We have sought to clarify the role of microsatellite instability (MSI) in pediatric versus adult HGG. MSI status was determined in 144 patients (71 pediatric and 73 adults) using a well established panel of five quasimonomorphic mononucleotide repeat markers. Expression of MLH1, MSH2, MSH6 and PMS2 was determined by immunohistochemistry, MLH1 was assessed for mutations by direct sequencing and promoter methylation using MS-PCR. DNA copy number profiles were derived using array CGH, and mutations in eighteen MSI target genes studied by multiplex PCR and genotyping. MSI was found in 14/71 (19.7%) pediatric cases, significantly more than observed in adults (5/73, 6.8%; p = 0.02, Chi-square test). MLH1 expression was downregulated in 10/13 cases, however no mutations or promoter methylation were found. MSH6 was absent in one pediatric MSI-High tumor, consistent with an inherited mismatch repair deficiency associated with germline MSH6 mutation. MSI was classed as Type A, and associated with a remarkably stable genomic profile. Of the eighteen classic MSI target genes, we identified mutations only in MSH6 and DNAPKcs and described a polymorphism in MRE11 without apparent functional consequences in DNA double strand break detection and repair. This study thus provides evidence for a potential novel molecular pathway in a proportion of gliomas associated with the presence of MSI. PMID- 21637785 TI - Promoter methylation in head and neck squamous cell carcinoma cell lines is significantly different than methylation in primary tumors and xenografts. AB - Studies designed to identify novel methylation events related to cancer often employ cancer cell lines in the discovery phase of the experiments and have a relatively low rate of discovery of cancer-related methylation events. An alternative algorithm for discovery of novel methylation in cancer uses primary tumor-derived xenografts instead of cell lines as the primary source of nucleic acid for evaluation. We evaluated DNA extracted from primary head and neck squamous cell carcinomas (HNSCC), xenografts grown from these primary tumors in nude mice, HNSCC-derived cell lines, normal oral mucosal samples, and minimally transformed oral keratinocyte-derived cell lines using Illumina Infinum Humanmethylation 27 genome-wide methylation microarrays. We found >2,200 statistically significant methylation differences between cancer cell lines and primary tumors and when comparing normal oral mucosa to keratinocyte cell lines. We found no statistically significant promoter methylation differences between primary tumor xenografts and primary tumors. This study demonstrates that tumor derived xenografts are highly accurate representations of promoter methylation in primary tumors and that cancer derived cell lines have significant drawbacks for discovery of promoter methylation alterations in primary tumors. These findings also support use of primary tumor xenografts for the study of methylation in cancer, drug discovery, and the development of personalized cancer treatments. PMID- 21637787 TI - Multi-scale approach for predicting fish species distributions across coral reef seascapes. AB - Two of the major limitations to effective management of coral reef ecosystems are a lack of information on the spatial distribution of marine species and a paucity of data on the interacting environmental variables that drive distributional patterns. Advances in marine remote sensing, together with the novel integration of landscape ecology and advanced niche modelling techniques provide an unprecedented opportunity to reliably model and map marine species distributions across many kilometres of coral reef ecosystems. We developed a multi-scale approach using three-dimensional seafloor morphology and across-shelf location to predict spatial distributions for five common Caribbean fish species. Seascape topography was quantified from high resolution bathymetry at five spatial scales (5-300 m radii) surrounding fish survey sites. Model performance and map accuracy was assessed for two high performing machine-learning algorithms: Boosted Regression Trees (BRT) and Maximum Entropy Species Distribution Modelling (MaxEnt). The three most important predictors were geographical location across the shelf, followed by a measure of topographic complexity. Predictor contribution differed among species, yet rarely changed across spatial scales. BRT provided 'outstanding' model predictions (AUC = >0.9) for three of five fish species. MaxEnt provided 'outstanding' model predictions for two of five species, with the remaining three models considered 'excellent' (AUC = 0.8-0.9). In contrast, MaxEnt spatial predictions were markedly more accurate (92% map accuracy) than BRT (68% map accuracy). We demonstrate that reliable spatial predictions for a range of key fish species can be achieved by modelling the interaction between the geographical location across the shelf and the topographic heterogeneity of seafloor structure. This multi-scale, analytic approach is an important new cost-effective tool to accurately delineate essential fish habitat and support conservation prioritization in marine protected area design, zoning in marine spatial planning, and ecosystem-based fisheries management. PMID- 21637786 TI - Nodal-dependent mesendoderm specification requires the combinatorial activities of FoxH1 and Eomesodermin. AB - Vertebrate mesendoderm specification requires the Nodal signaling pathway and its transcriptional effector FoxH1. However, loss of FoxH1 in several species does not reliably cause the full range of loss-of-Nodal phenotypes, indicating that Nodal signals through additional transcription factors during early development. We investigated the FoxH1-dependent and -independent roles of Nodal signaling during mesendoderm patterning using a novel recessive zebrafish FoxH1 mutation called midway, which produces a C-terminally truncated FoxH1 protein lacking the Smad-interaction domain but retaining DNA-binding capability. Using a combination of gel shift assays, Nodal overexpression experiments, and genetic epistasis analyses, we demonstrate that midway more accurately represents a complete loss of FoxH1-dependent Nodal signaling than the existing zebrafish FoxH1 mutant schmalspur. Maternal-zygotic midway mutants lack notochords, in agreement with FoxH1 loss in other organisms, but retain near wild-type expression of markers of endoderm and various nonaxial mesoderm fates, including paraxial and intermediate mesoderm and blood precursors. We found that the activity of the T-box transcription factor Eomesodermin accounts for specification of these tissues in midway embryos. Inhibition of Eomesodermin in midway mutants severely reduces the specification of these tissues and effectively phenocopies the defects seen upon complete loss of Nodal signaling. Our results indicate that the specific combinations of transcription factors available for signal transduction play critical and separable roles in determining Nodal pathway output during mesendoderm patterning. Our findings also offer novel insights into the co evolution of the Nodal signaling pathway, the notochord specification program, and the chordate branch of the deuterostome family of animals. PMID- 21637784 TI - Association of genetic variants in complement factor H and factor H-related genes with systemic lupus erythematosus susceptibility. AB - Systemic lupus erythematosus (SLE), a complex polygenic autoimmune disease, is associated with increased complement activation. Variants of genes encoding complement regulator factor H (CFH) and five CFH-related proteins (CFHR1-CFHR5) within the chromosome 1q32 locus linked to SLE, have been associated with multiple human diseases and may contribute to dysregulated complement activation predisposing to SLE. We assessed 60 SNPs covering the CFH-CFHRs region for association with SLE in 15,864 case-control subjects derived from four ethnic groups. Significant allelic associations with SLE were detected in European Americans (EA) and African Americans (AA), which could be attributed to an intronic CFH SNP (rs6677604, in intron 11, P(meta) = 6.6*10(-8), OR = 1.18) and an intergenic SNP between CFHR1 and CFHR4 (rs16840639, P(meta) = 2.9*10(-7), OR = 1.17) rather than to previously identified disease-associated CFH exonic SNPs, including I62V, Y402H, A474A, and D936E. In addition, allelic association of rs6677604 with SLE was subsequently confirmed in Asians (AS). Haplotype analysis revealed that the underlying causal variant, tagged by rs6677604 and rs16840639, was localized to a ~146 kb block extending from intron 9 of CFH to downstream of CFHR1. Within this block, the deletion of CFHR3 and CFHR1 (CFHR3-1Delta), a likely causal variant measured using multiplex ligation-dependent probe amplification, was tagged by rs6677604 in EA and AS and rs16840639 in AA, respectively. Deduced from genotypic associations of tag SNPs in EA, AA, and AS, homozygous deletion of CFHR3-1Delta (P(meta) = 3.2*10(-7), OR = 1.47) conferred a higher risk of SLE than heterozygous deletion (P(meta) = 3.5*10(-4), OR = 1.14). These results suggested that the CFHR3-1Delta deletion within the SLE-associated block, but not the previously described exonic SNPs of CFH, might contribute to the development of SLE in EA, AA, and AS, providing new insights into the role of complement regulators in the pathogenesis of SLE. PMID- 21637788 TI - Integrated genome-scale prediction of detrimental mutations in transcription networks. AB - A central challenge in genetics is to understand when and why mutations alter the phenotype of an organism. The consequences of gene inhibition have been systematically studied and can be predicted reasonably well across a genome. However, many sequence variants important for disease and evolution may alter gene regulation rather than gene function. The consequences of altering a regulatory interaction (or "edge") rather than a gene (or "node") in a network have not been as extensively studied. Here we use an integrative analysis and evolutionary conservation to identify features that predict when the loss of a regulatory interaction is detrimental in the extensively mapped transcription network of budding yeast. Properties such as the strength of an interaction, location and context in a promoter, regulator and target gene importance, and the potential for compensation (redundancy) associate to some extent with interaction importance. Combined, however, these features predict quite well whether the loss of a regulatory interaction is detrimental across many promoters and for many different transcription factors. Thus, despite the potential for regulatory diversity, common principles can be used to understand and predict when changes in regulation are most harmful to an organism. PMID- 21637789 TI - A novel mouse synaptonemal complex protein is essential for loading of central element proteins, recombination, and fertility. AB - The synaptonemal complex (SC) is a proteinaceous, meiosis-specific structure that is highly conserved in evolution. During meiosis, the SC mediates synapsis of homologous chromosomes. It is essential for proper recombination and segregation of homologous chromosomes, and therefore for genome haploidization. Mutations in human SC genes can cause infertility. In order to gain a better understanding of the process of SC assembly in a model system that would be relevant for humans, we are investigating meiosis in mice. Here, we report on a newly identified component of the murine SC, which we named SYCE3. SYCE3 is strongly conserved among mammals and localizes to the central element (CE) of the SC. By generating a Syce3 knockout mouse, we found that SYCE3 is required for fertility in both sexes. Loss of SYCE3 blocks synapsis initiation and results in meiotic arrest. In the absence of SYCE3, initiation of meiotic recombination appears to be normal, but its progression is severely impaired resulting in complete absence of MLH1 foci, which are presumed markers of crossovers in wild-type meiocytes. In the process of SC assembly, SYCE3 is required downstream of transverse filament protein SYCP1, but upstream of the other previously described CE-specific proteins. We conclude that SYCE3 enables chromosome loading of the other CE specific proteins, which in turn would promote synapsis between homologous chromosomes. PMID- 21637790 TI - Development of resistance towards artesunate in MDA-MB-231 human breast cancer cells. AB - Breast cancer is the most common cancer and the second leading cause of cancer death in industrialized countries. Systemic treatment of breast cancer is effective at the beginning of therapy. However, after a variable period of time, progression occurs due to therapy resistance. Artesunate, clinically used as anti malarial agent, has recently revealed remarkable anti-tumor activity offering a role as novel candidate for cancer chemotherapy. We analyzed the anti-tumor effects of artesunate in metastasizing breast carcinoma in vitro and in vivo. Unlike as expected, artesunate induced resistance in highly metastatic human breast cancer cells MDA-MB-231. Likewise acquired resistance led to abolishment of apoptosis and cytotoxicity in pre-treated MDA-MB-231 cells. In contrast, artesunate was more cytotoxic towards the less tumorigenic MDA-MB-468 cells without showing resistance. Unraveling the underlying molecular mechanisms, we found that resistance was induced due to activation of the tumor progression related transcription factors NFkappaB and AP-1. Thereby transcription, expression and activity of the matrix-degrading enzyme MMP-1, whose function is correlated with increased invasion and metastasis, was up-regulated upon acquisition of resistance. Additionally, activation of the apoptosis-related factor NFkappaB lead to increased expression of ant-apoptotic bcl2 and reduced expression of pro-apoptotic bax. Application of artesunate in vivo in a model of xenografted breast cancer showed, that tumors growth was not efficiently abolished as compared to the control drug doxorubicin. Taken together our in vitro and in vivo results correlate well showing for the first time that artesunate induces resistance in highly metastatic breast tumors. PMID- 21637791 TI - Meiotic recombination intermediates are resolved with minimal crossover formation during return-to-growth, an analogue of the mitotic cell cycle. AB - Accurate segregation of homologous chromosomes of different parental origin (homologs) during the first division of meiosis (meiosis I) requires inter homolog crossovers (COs). These are produced at the end of meiosis I prophase, when recombination intermediates that contain Holliday junctions (joint molecules, JMs) are resolved, predominantly as COs. JM resolution during the mitotic cell cycle is less well understood, mainly due to low levels of inter homolog JMs. To compare JM resolution during meiosis and the mitotic cell cycle, we used a unique feature of Saccharomyces cerevisiae, return to growth (RTG), where cells undergoing meiosis can be returned to the mitotic cell cycle by a nutritional shift. By performing RTG with ndt80 mutants, which arrest in meiosis I prophase with high levels of interhomolog JMs, we could readily monitor JM resolution during the first cell division of RTG genetically and, for the first time, at the molecular level. In contrast to meiosis, where most JMs resolve as COs, most JMs were resolved during the first 1.5-2 hr after RTG without producing COs. Subsequent resolution of the remaining JMs produced COs, and this CO production required the Mus81/Mms4 structure-selective endonuclease. RTG in sgs1 DeltaC795 mutants, which lack the helicase and Holliday junction-binding domains of this BLM homolog, led to a substantial delay in JM resolution; and subsequent JM resolution produced both COs and NCOs. Based on these findings, we suggest that most JMs are resolved during the mitotic cell cycle by dissolution, an Sgs1 helicase-dependent process that produces only NCOs. JMs that escape dissolution are mostly resolved by Mus81/Mms4-dependent cleavage that produces both COs and NCOs in a relatively unbiased manner. Thus, in contrast to meiosis, where JM resolution is heavily biased towards COs, JM resolution during RTG minimizes CO formation, thus maintaining genome integrity and minimizing loss of heterozygosity. PMID- 21637792 TI - A genetic and structural study of genome rearrangements mediated by high copy repeat Ty1 elements. AB - Ty elements are high copy number, dispersed repeated sequences in the Saccharomyces cerevisiae genome known to mediate gross chromosomal rearrangements (GCRs). Here we found that introduction of Ty912, a previously identified Ty1 element, onto the non-essential terminal region of the left arm of chromosome V led to a 380-fold increase in the rate of accumulating GCRs in a wild-type strain. A survey of 48 different mutations identified those that either increased or decreased the rate of Ty-mediated GCRs and demonstrated that suppression of Ty mediated GCRs differs from that of both low copy repeat sequence- and single copy sequence-mediated GCRs. The majority of the Ty912-mediated GCRs observed were monocentric nonreciprocal translocations mediated by RAD52-dependent homologous recombination (HR) between Ty912 and a Ty element on another chromosome arm. The remaining Ty912-mediated GCRs appeared to involve Ty912-mediated formation of unstable dicentric translocation chromosomes that were resolved by one or more Ty mediated breakage-fusion-bridge cycles. Overall, the results demonstrate that the Ty912-mediated GCR assay is an excellent model for understanding mechanisms and pathways that suppress genome rearrangements mediated by high copy number repeat sequences, as well as the mechanisms by which such rearrangements occur. PMID- 21637793 TI - Structural and functional differences in the long non-coding RNA hotair in mouse and human. AB - Long non-coding RNAs regulate various biological processes such as dosage compensation, imprinting, and chromatin organization. HOTAIR, a paradigm of this new class of RNAs, is localized within the human HOXC gene cluster and was shown, in human cells, to regulate HOXD genes in trans via the recruitment of Polycomb Repressive Complex 2 (PRC2), followed by the trimethylation of lysine 27 of histone H3. We looked for the presence of Hotair in mice to assess whether this in trans mechanism was conserved, in particular at the developmental stages, when Hoxd genes must be tightly regulated. We show that the cognate mouse Hotair is poorly conserved in sequence; and its absence, along with the deletion of the HoxC cluster, has surprisingly little effect in vivo, neither on the expression pattern or transcription efficiency, nor on the amount of K27me3 coverage of different Hoxd target genes. We conclude that Hotair may have rapidly evolved within mammals and acquired a functional importance in humans that is not easily revealed in mice. Alternatively, redundant or compensatory mechanisms may mask its function when studied under physiological conditions. PMID- 21637794 TI - Identification, replication, and functional fine-mapping of expression quantitative trait loci in primary human liver tissue. AB - The discovery of expression quantitative trait loci ("eQTLs") can help to unravel genetic contributions to complex traits. We identified genetic determinants of human liver gene expression variation using two independent collections of primary tissue profiled with Agilent (n = 206) and Illumina (n = 60) expression arrays and Illumina SNP genotyping (550K), and we also incorporated data from a published study (n = 266). We found that ~30% of SNP-expression correlations in one study failed to replicate in either of the others, even at thresholds yielding high reproducibility in simulations, and we quantified numerous factors affecting reproducibility. Our data suggest that drug exposure, clinical descriptors, and unknown factors associated with tissue ascertainment and analysis have substantial effects on gene expression and that controlling for hidden confounding variables significantly increases replication rate. Furthermore, we found that reproducible eQTL SNPs were heavily enriched near gene starts and ends, and subsequently resequenced the promoters and 3'UTRs for 14 genes and tested the identified haplotypes using luciferase assays. For three genes, significant haplotype-specific in vitro functional differences correlated directly with expression levels, suggesting that many bona fide eQTLs result from functional variants that can be mechanistically isolated in a high-throughput fashion. Finally, given our study design, we were able to discover and validate hundreds of liver eQTLs. Many of these relate directly to complex traits for which liver-specific analyses are likely to be relevant, and we identified dozens of potential connections with disease-associated loci. These included previously characterized eQTL contributors to diabetes, drug response, and lipid levels, and they suggest novel candidates such as a role for NOD2 expression in leprosy risk and C2orf43 in prostate cancer. In general, the work presented here will be valuable for future efforts to precisely identify and functionally characterize genetic contributions to a variety of complex traits. PMID- 21637795 TI - Simulation of the undiseased human cardiac ventricular action potential: model formulation and experimental validation. AB - Cellular electrophysiology experiments, important for understanding cardiac arrhythmia mechanisms, are usually performed with channels expressed in non myocytes, or with non-human myocytes. Differences between cell types and species affect results. Thus, an accurate model for the undiseased human ventricular action potential (AP) which reproduces a broad range of physiological behaviors is needed. Such a model requires extensive experimental data, but essential elements have been unavailable. Here, we develop a human ventricular AP model using new undiseased human ventricular data: Ca(2+) versus voltage dependent inactivation of L-type Ca(2+) current (I(CaL)); kinetics for the transient outward, rapid delayed rectifier (I(Kr)), Na(+)/Ca(2+) exchange (I(NaCa)), and inward rectifier currents; AP recordings at all physiological cycle lengths; and rate dependence and restitution of AP duration (APD) with and without a variety of specific channel blockers. Simulated APs reproduced the experimental AP morphology, APD rate dependence, and restitution. Using undiseased human mRNA and protein data, models for different transmural cell types were developed. Experiments for rate dependence of Ca(2+) (including peak and decay) and intracellular sodium ([Na(+)](i)) in undiseased human myocytes were quantitatively reproduced by the model. Early afterdepolarizations were induced by I(Kr) block during slow pacing, and AP and Ca(2+) alternans appeared at rates >200 bpm, as observed in the nonfailing human ventricle. Ca(2+)/calmodulin dependent protein kinase II (CaMK) modulated rate dependence of Ca(2+) cycling. I(NaCa) linked Ca(2+) alternation to AP alternans. CaMK suppression or SERCA upregulation eliminated alternans. Steady state APD rate dependence was caused primarily by changes in [Na(+)](i), via its modulation of the electrogenic Na(+)/K(+) ATPase current. At fast pacing rates, late Na(+) current and I(CaL) were also contributors. APD shortening during restitution was primarily dependent on reduced late Na(+) and I(CaL) currents due to inactivation at short diastolic intervals, with additional contribution from elevated I(Kr) due to incomplete deactivation. PMID- 21637796 TI - The ISWI chromatin remodeler organizes the hsromega ncRNA-containing omega speckle nuclear compartments. AB - The complexity in composition and function of the eukaryotic nucleus is achieved through its organization in specialized nuclear compartments. The Drosophila chromatin remodeling ATPase ISWI plays evolutionarily conserved roles in chromatin organization. Interestingly, ISWI genetically interacts with the hsromega gene, encoding multiple non-coding RNAs (ncRNA) essential, among other functions, for the assembly and organization of the omega speckles. The nucleoplasmic omega speckles play important functions in RNA metabolism, in normal and stressed cells, by regulating availability of hnRNPs and some other RNA processing proteins. Chromatin remodelers, as well as nuclear speckles and their associated ncRNAs, are emerging as important components of gene regulatory networks, although their functional connections have remained poorly defined. Here we provide multiple lines of evidence showing that the hsromega ncRNA interacts in vivo and in vitro with ISWI, regulating its ATPase activity. Remarkably, we found that the organization of nucleoplasmic omega speckles depends on ISWI function. Our findings highlight a novel role for chromatin remodelers in organization of nucleoplasmic compartments, providing the first example of interaction between an ATP-dependent chromatin remodeler and a large ncRNA. PMID- 21637797 TI - Pathway analysis of expression data: deciphering functional building blocks of complex diseases. PMID- 21637798 TI - An acidic loop and cognate phosphorylation sites define a molecular switch that modulates ubiquitin charging activity in Cdc34-like enzymes. AB - E2 ubiquitin-conjugating enzymes are crucial mediators of protein ubiquitination, which strongly influence the ultimate fate of the target substrates. Recently, it has been shown that the activity of several enzymes of the ubiquitination pathway is finely tuned by phosphorylation, an ubiquitous mechanism for cellular regulation, which modulates protein conformation. In this contribution, we provide the first rationale, at the molecular level, of the regulatory mechanism mediated by casein kinase 2 (CK2) phosphorylation of E2 Cdc34-like enzymes. In particular, we identify two co-evolving signature elements in one of the larger families of E2 enzymes: an acidic insertion in beta4alpha2 loop in the proximity of the catalytic cysteine and two conserved key serine residues within the catalytic domain, which are phosphorylated by CK2. Our investigations, using yeast Cdc34 as a model, through 2.5 us molecular dynamics simulations and biochemical assays, define these two elements as an important phosphorylation controlled switch that modulates opening and closing of the catalytic cleft. The mechanism relies on electrostatic repulsions between a conserved serine phosphorylated by CK2 and the acidic residues of the beta4alpha2 loop, promoting E2 ubiquitin charging activity. Our investigation identifies a new and unexpected pivotal role for the acidic loop, providing the first evidence that this loop is crucial not only for downstream events related to ubiquitin chain assembly, but is also mandatory for the modulation of an upstream crucial step of the ubiquitin pathway: the ubiquitin charging in the E2 catalytic cleft. PMID- 21637799 TI - A role for both conformational selection and induced fit in ligand binding by the LAO protein. AB - Molecular recognition is determined by the structure and dynamics of both a protein and its ligand, but it is difficult to directly assess the role of each of these players. In this study, we use Markov State Models (MSMs) built from atomistic simulations to elucidate the mechanism by which the Lysine-, Arginine-, Ornithine-binding (LAO) protein binds to its ligand. We show that our model can predict the bound state, binding free energy, and association rate with reasonable accuracy and then use the model to dissect the binding mechanism. In the past, this binding event has often been assumed to occur via an induced fit mechanism because the protein's binding site is completely closed in the bound state, making it impossible for the ligand to enter the binding site after the protein has adopted the closed conformation. More complex mechanisms have also been hypothesized, but these have remained controversial. Here, we are able to directly observe roles for both the conformational selection and induced fit mechanisms in LAO binding. First, the LAO protein tends to form a partially closed encounter complex via conformational selection (that is, the apo protein can sample this state), though the induced fit mechanism can also play a role here. Then, interactions with the ligand can induce a transition to the bound state. Based on these results, we propose that MSMs built from atomistic simulations may be a powerful way of dissecting ligand-binding mechanisms and may eventually facilitate a deeper understanding of allostery as well as the prediction of new protein-ligand interactions, an important step in drug discovery. PMID- 21637800 TI - Ten simple rules for providing a scientific Web resource. PMID- 21637801 TI - A zebrafish model of Roberts syndrome reveals that Esco2 depletion interferes with development by disrupting the cell cycle. AB - The human developmental diseases Cornelia de Lange Syndrome (CdLS) and Roberts Syndrome (RBS) are both caused by mutations in proteins responsible for sister chromatid cohesion. Cohesion is mediated by a multi-subunit complex called cohesin, which is loaded onto chromosomes by NIPBL. Once on chromosomes, cohesin binding is stabilized in S phase upon acetylation by ESCO2. CdLS is caused by heterozygous mutations in NIPBL or cohesin subunits SMC1A and SMC3, and RBS is caused by homozygous mutations in ESCO2. The genetic cause of both CdLS and RBS reside within the chromosome cohesion apparatus, and therefore they are collectively known as "cohesinopathies". However, the two syndromes have distinct phenotypes, with differences not explained by their shared ontology. In this study, we have used the zebrafish model to distinguish between developmental pathways downstream of cohesin itself, or its acetylase ESCO2. Esco2 depleted zebrafish embryos exhibit features that resemble RBS, including mitotic defects, craniofacial abnormalities and limb truncations. A microarray analysis of Esco2 depleted embryos revealed that different subsets of genes are regulated downstream of Esco2 when compared with cohesin subunit Rad21. Genes downstream of Rad21 showed significant enrichment for transcriptional regulators, while Esco2 regulated genes were more likely to be involved the cell cycle or apoptosis. RNA in situ hybridization showed that runx1, which is spatiotemporally regulated by cohesin, is expressed normally in Esco2-depleted embryos. Furthermore, myca, which is downregulated in rad21 mutants, is upregulated in Esco2-depleted embryos. High levels of cell death contributed to the morphology of Esco2 depleted embryos without affecting specific developmental pathways. We propose that cell proliferation defects and apoptosis could be the primary cause of the features of RBS. Our results show that mutations in different elements of the cohesion apparatus have distinct developmental outcomes, and provide insight into why CdLS and RBS are distinct diseases. PMID- 21637802 TI - Cross-sectional analysis of late HAART initiation in Latin America and the Caribbean: late testers and late presenters. AB - BACKGROUND: Starting HAART in a very advanced stage of disease is assumed to be the most prevalent form of initiation in HIV-infected subjects in developing countries. Data from Latin America and the Caribbean is still lacking. Our main objective was to determine the frequency, risk factors and trends in time for being late HAART initiator (LHI) in this region. METHODOLOGY: Cross-sectional analysis from 9817 HIV-infected treatment-naive patients initiating HAART at 6 sites (Argentina, Chile, Haiti, Honduras, Peru and Mexico) from October 1999 to July 2010. LHI had CD4(+) count <=200 cells/mm(3) prior to HAART. Late testers (LT) were those LHI who initiated HAART within 6 months of HIV diagnosis. Late presenters (LP) initiated after 6 months of diagnosis. Prevalence, risk factors and trends over time were analyzed. PRINCIPAL FINDINGS: Among subjects starting HAART (n = 9817) who had baseline CD4(+) available (n = 8515), 76% were LHI: Argentina (56%[95%CI:52-59]), Chile (80%[95%CI:77-82]), Haiti (76%[95%CI:74-77]), Honduras (91%[95%CI:87-94]), Mexico (79%[95%CI:75-83]), Peru (86%[95%CI:84-88]). The proportion of LHI statistically changed over time (except in Honduras) (p<=0.02; Honduras p = 0.7), with a tendency towards lower rates in recent years. Males had increased risk of LHI in Chile, Haiti, Peru, and in the combined site analyses (CSA). Older patients were more likely LHI in Argentina and Peru (OR 1.21 per +10-year of age, 95%CI:1.02-1.45; OR 1.20, 95%CI:1.02-1.43; respectively), but not in CSA (OR 1.07, 95%CI:0.94-1.21). Higher education was associated with decreased risk for LHI in Chile (OR 0.92 per +1-year of education, 95%CI:0.87-0.98) (similar trends in Mexico, Peru, and CSA). LHI with date of HIV-diagnosis available, 55% were LT and 45% LP. CONCLUSION: LHI was highly prevalent in CCASAnet sites, mostly due to LT; the main risk factors associated were being male and older age. Earlier HIV-diagnosis and earlier treatment initiation are needed to maximize benefits from HAART in the region. PMID- 21637804 TI - Distinct control of initiation and metrics of memory-guided saccades and vergence by the FEF: a TMS study. AB - BACKGROUND: The initiation of memory guided saccades is known to be controlled by the frontal eye field (FEF). Recent physiological studies showed the existence of an area close to FEF that controls also vergence initiation and execution. This study is to explore the effect of transcranial magnetic simulation (TMS) over FEF on the control of memory-guided saccade-vergence eye movements. METHODOLOGY/PRINCIPAL FINDINGS: Subjects had to make an eye movement in dark towards a target flashed 1 sec earlier (memory delay); the location of the target relative to fixation point was such as to require either a vergence along the median plane, or a saccade, or a saccade with vergence; trials were interleaved. Single pulse TMS was applied on the left or right FEF; it was delivered at 100 ms after the end of memory delay, i.e. extinction of fixation LED that was the "go" signal. Twelve healthy subjects participated in the study. TMS of left or right FEF prolonged the latency of all types of eye movements; the increase varied from 21 to 56 ms and was particularly strong for the divergence movements. This indicates that FEF is involved in the initiation of all types of memory guided movement in the 3D space. TMS of the FEF also altered the accuracy but only for leftward saccades combined with either convergence or divergence; intrasaccadic vergence also increased after TMS of the FEF. CONCLUSIONS/SIGNIFICANCE: The results suggest anisotropy in the quality of space memory and are discussed in the context of other known perceptual motor anisotropies. PMID- 21637803 TI - Exonic DNA sequencing of ERBB4 in bipolar disorder. AB - The Neuregulin-ErbB4 pathway plays a crucial role in brain development and constitutes one of the most biologically plausible signaling pathways implicated in schizophrenia and, to a lesser extent, in bipolar disorder (BP). However, recent genome-wide association analyses have not provided evidence for common variation in NRG1 or ERBB4 influencing schizophrenia or bipolar disorder susceptibility. In this study, we investigate the role of rare coding variants in ERBB4 in BP cases with mood-incongruent psychotic features, a form of BP with arguably the greatest phenotypic overlap with schizophrenia. We performed Sanger sequencing of all 28 exons in ERBB4, as well as part of the promoter and part of the 3'UTR sequence, hypothesizing that rare deleterious variants would be found in 188 cases with mood-incongruent psychosis from the GAIN BP study. We found 42 variants, of which 16 were novel, although none were non-synonymous or clearly deleterious. One of the novel variants, present in 11.2% of cases, is located next to an alternative stop codon, which is associated with a shortened transcript of ERBB4 that is not translated. We genotyped this variant in the GAIN BP case-control samples and found a marginally significant association with mood incongruent psychotic BP compared with controls (additive model: OR = 1.64, P value = 0.055; dominant model: OR = 1.73. P-value = 0.039). In conclusion, we found no rare variants of clear deleterious effect, but did uncover a modestly associated novel variant that could affect alternative splicing of ERBB4. However, the modest sample size in this study cannot definitively rule out a role for rare variants in bipolar disorder and studies with larger sample sizes are needed to confirm the observed association. PMID- 21637805 TI - Host-pathogen o-methyltransferase similarity and its specific presence in highly virulent strains of Francisella tularensis suggests molecular mimicry. AB - Whole genome comparative studies of many bacterial pathogens have shown an overall high similarity of gene content (>95%) between phylogenetically distinct subspecies. In highly clonal species that share the bulk of their genomes subtle changes in gene content and small-scale polymorphisms, especially those that may alter gene expression and protein-protein interactions, are more likely to have a significant effect on the pathogen's biology. In order to better understand molecular attributes that may mediate the adaptation of virulence in infectious bacteria, a comparative study was done to further analyze the evolution of a gene encoding an o-methyltransferase that was previously identified as a candidate virulence factor due to its conservation specifically in highly pathogenic Francisella tularensis subsp. tularensis strains. The o-methyltransferase gene is located in the genomic neighborhood of a known pathogenicity island and predicted site of rearrangement. Distinct o-methyltransferase subtypes are present in different Francisella tularensis subspecies. Related protein families were identified in several host species as well as species of pathogenic bacteria that are otherwise very distant phylogenetically from Francisella, including species of Mycobacterium. A conserved sequence motif profile is present in the mammalian host and pathogen protein sequences, and sites of non-synonymous variation conserved in Francisella subspecies specific o-methyltransferases map proximally to the predicted active site of the orthologous human protein structure. Altogether, evidence suggests a role of the F. t. subsp. tularensis protein in a mechanism of molecular mimicry, similar perhaps to Legionella and Coxiella. These findings therefore provide insights into the evolution of niche-restriction and virulence in Francisella, and have broader implications regarding the molecular mechanisms that mediate host-pathogen relationships. PMID- 21637806 TI - Isolation of thylakoid membrane complexes from rice by a new double-strips BN/SDS PAGE and bioinformatics prediction of stromal ridge subunits interaction. AB - Thylakoid membrane complexes of rice (Oryza sativa L.) play crucial roles in growth and crop production. Understanding of protein interactions within the complex would provide new insights into photosynthesis. Here, a new "Double Strips BN/SDS-PAGE" method was employed to separate thylakoid membrane complexes in order to increase the protein abundance on 2D-gels and to facilitate the identification of hydrophobic transmembrane proteins. A total of 58 protein spots could be observed and subunit constitution of these complexes exhibited on 2D gels. The generality of this new approach was confirmed using thylakoid membrane from spinach (Spinacia oleracea) and pumpkin (Cucurita spp). Furthermore, the proteins separated from rice thylakoid membrane were identified by the mass spectrometry (MS). The stromal ridge proteins PsaD and PsaE were identified both in the holo- and core- PSI complexes of rice. Using molecular dynamics simulation to explore the recognition mechanism of these subunits, we showed that salt bridge interactions between residues R19 of PsaC and E168 of PasD as well as R75 of PsaC and E91 of PsaD played important roles in the stability of the complex. This stromal ridge subunits interaction was also supported by the subsequent analysis of the binding free energy, the intramolecular distances and the intramolecular energy. PMID- 21637807 TI - New classes of alanine racemase inhibitors identified by high-throughput screening show antimicrobial activity against Mycobacterium tuberculosis. AB - BACKGROUND: In an effort to discover new drugs to treat tuberculosis (TB) we chose alanine racemase as the target of our drug discovery efforts. In Mycobacterium tuberculosis, the causative agent of TB, alanine racemase plays an essential role in cell wall synthesis as it racemizes L-alanine into D-alanine, a key building block in the biosynthesis of peptidoglycan. Good antimicrobial effects have been achieved by inhibition of this enzyme with suicide substrates, but the clinical utility of this class of inhibitors is limited due to their lack of target specificity and toxicity. Therefore, inhibitors that are not substrate analogs and that act through different mechanisms of enzyme inhibition are necessary for therapeutic development for this drug target. METHODOLOGY/PRINCIPAL FINDINGS: To obtain non-substrate alanine racemase inhibitors, we developed a high-throughput screening platform and screened 53,000 small molecule compounds for enzyme-specific inhibitors. We examined the 'hits' for structural novelty, antimicrobial activity against M. tuberculosis, general cellular cytotoxicity, and mechanism of enzyme inhibition. We identified seventeen novel non-substrate alanine racemase inhibitors that are structurally different than any currently known enzyme inhibitors. Seven of these are active against M. tuberculosis and minimally cytotoxic against mammalian cells. CONCLUSIONS/SIGNIFICANCE: This study highlights the feasibility of obtaining novel alanine racemase inhibitor lead compounds by high-throughput screening for development of new anti-TB agents. PMID- 21637808 TI - Lymphoadenopathy during lyme borreliosis is caused by spirochete migration induced specific B cell activation. AB - Lymphadenopathy is a hallmark of acute infection with Borrelia burgdorferi, a tick-borne spirochete and causative agent of Lyme borreliosis, but the underlying causes and the functional consequences of this lymph node enlargement have not been revealed. The present study demonstrates that extracellular, live spirochetes accumulate in the cortical areas of lymph nodes following infection of mice with either host-adapted, or tick-borne B. burgdorferi and that they, but not inactivated spirochetes, drive the lymphadenopathy. The ensuing lymph node response is characterized by strong, rapid extrafollicular B cell proliferation and differentiation to plasma cells, as assessed by immunohistochemistry, flow cytometry and ELISPOT analysis, while germinal center reactions were not consistently observed. The extrafollicular nature of this B cell response and its strongly IgM-skewed isotype profile bear the hallmarks of a T-independent response. The induced B cell response does appear, however, to be largely antigen specific. Use of a cocktail of recombinant, in vivo-expressed B. burgdorferi antigens revealed the robust induction of borrelia-specific antibody-secreting cells by ELISPOT. Furthermore, nearly a quarter of hybridomas generated from regional lymph nodes during acute infection showed reactivity against a small number of recombinant Borrelia-antigens. Finally, neither the quality nor the magnitude of the B cell responses was altered in mice lacking the Toll-like receptor adaptor molecule MyD88. Together, these findings suggest a novel evasion strategy for B. burgdorferi: subversion of the quality of a strongly induced, potentially protective borrelia-specific antibody response via B. burdorferi's accumulation in lymph nodes. PMID- 21637809 TI - Acquisition of human-type receptor binding specificity by new H5N1 influenza virus sublineages during their emergence in birds in Egypt. AB - Highly pathogenic avian influenza A virus subtype H5N1 is currently widespread in Asia, Europe, and Africa, with 60% mortality in humans. In particular, since 2009 Egypt has unexpectedly had the highest number of human cases of H5N1 virus infection, with more than 50% of the cases worldwide, but the basis for this high incidence has not been elucidated. A change in receptor binding affinity of the viral hemagglutinin (HA) from alpha2,3- to alpha2,6-linked sialic acid (SA) is thought to be necessary for H5N1 virus to become pandemic. In this study, we conducted a phylogenetic analysis of H5N1 viruses isolated between 2006 and 2009 in Egypt. The phylogenetic results showed that recent human isolates clustered disproportionally into several new H5 sublineages suggesting that their HAs have changed their receptor specificity. Using reverse genetics, we found that these H5 sublineages have acquired an enhanced binding affinity for alpha2,6 SA in combination with residual affinity for alpha2,3 SA, and identified the amino acid mutations that produced this new receptor specificity. Recombinant H5N1 viruses with a single mutation at HA residue 192 or a double mutation at HA residues 129 and 151 had increased attachment to and infectivity in the human lower respiratory tract but not in the larynx. These findings correlated with enhanced virulence of the mutant viruses in mice. Interestingly, these H5 viruses, with increased affinity to alpha2,6 SA, emerged during viral diversification in bird populations and subsequently spread to humans. Our findings suggested that emergence of new H5 sublineages with alpha2,6 SA specificity caused a subsequent increase in human H5N1 influenza virus infections in Egypt, and provided data for understanding the virus's pandemic potential. PMID- 21637810 TI - Population structure of invasive Streptococcus pneumoniae in The Netherlands in the pre-vaccination era assessed by MLVA and capsular sequence typing. AB - The introduction of nationwide pneumococcal vaccination may lead to serotype replacement and the emergence of new variants that have expanded their genetic repertoire through recombination. To monitor alterations in the pneumococcal population structure, we have developed and utilized Capsular Sequence Typing (CST) in addition to Multiple-Locus Variable number tandem repeat Analysis (MLVA).To assess the serotype of each isolate CST was used. Based on the determination of the partial sequence of the capsular wzh gene, this method assigns a capsular type of an isolate within a single PCR reaction using multiple primersets. The genetic background of pneumococcal isolates was assessed by MLVA. MLVA and CST were used to create a snapshot of the Dutch pneumococcal population causing invasive disease before the introduction of the 7-valent pneumococcal conjugate vaccine in The Netherlands in 2006. A total of 1154 clinical isolates collected and serotyped by the Netherlands Reference Laboratory for Bacterial Meningitis were included in the snapshot. The CST was successful in discriminating most serotypes present in our collection. MLVA demonstrated that isolates belonging to some serotypes had a relatively high genetic diversity whilst other serotypes had a very homogeneous genetic background. MLVA and CST appear to be valuable tools to determine the population structure of pneumococcal isolates and are useful in monitoring the effects of pneumococcal vaccination. PMID- 21637811 TI - Suboptimal activation of antigen-specific CD4+ effector cells enables persistence of M. tuberculosis in vivo. AB - Adaptive immunity to Mycobacterium tuberculosis controls progressive bacterial growth and disease but does not eradicate infection. Among CD4+ T cells in the lungs of M. tuberculosis-infected mice, we observed that few produced IFN-gamma without ex vivo restimulation. Therefore, we hypothesized that one mechanism whereby M. tuberculosis avoids elimination is by limiting activation of CD4+ effector T cells at the site of infection in the lungs. To test this hypothesis, we adoptively transferred Th1-polarized CD4+ effector T cells specific for M. tuberculosis Ag85B peptide 25 (P25TCRTh1 cells), which trafficked to the lungs of infected mice and exhibited antigen-dependent IFN-gamma production. During the early phase of infection, ~10% of P25TCRTh1 cells produced IFN-gamma in vivo; this declined to <1% as infection progressed to chronic phase. Bacterial downregulation of fbpB (encoding Ag85B) contributed to the decrease in effector T cell activation in the lungs, as a strain of M. tuberculosis engineered to express fbpB in the chronic phase stimulated P25TCRTh1 effector cells at higher frequencies in vivo, and this resulted in CD4+ T cell-dependent reduction of lung bacterial burdens and prolonged survival of mice. Administration of synthetic peptide 25 alone also increased activation of endogenous antigen-specific effector cells and reduced the bacterial burden in the lungs without apparent host toxicity. These results indicate that CD4+ effector T cells are activated at suboptimal frequencies in tuberculosis, and that increasing effector T cell activation in the lungs by providing one or more epitope peptides may be a successful strategy for TB therapy. PMID- 21637812 TI - Distribution of attention modulates salience signals in early visual cortex. AB - Previous research has shown that the extent to which people spread attention across the visual field plays a crucial role in visual selection and the occurrence of bottom-up driven attentional capture. Consistent with previous findings, we show that when attention was diffusely distributed across the visual field while searching for a shape singleton, an irrelevant salient color singleton captured attention. However, while using the very same displays and task, no capture was observed when observers initially focused their attention at the center of the display. Using event-related fMRI, we examined the modulation of retinotopic activity related to attentional capture in early visual areas. Because the sensory display characteristics were identical in both conditions, we were able to isolate the brain activity associated with exogenous attentional capture. The results show that spreading of attention leads to increased bottom up exogenous capture and increased activity in visual area V3 but not in V2 and V1. PMID- 21637813 TI - Crystal structure and functional analysis of the SARS-coronavirus RNA cap 2'-O methyltransferase nsp10/nsp16 complex. AB - Cellular and viral S-adenosylmethionine-dependent methyltransferases are involved in many regulated processes such as metabolism, detoxification, signal transduction, chromatin remodeling, nucleic acid processing, and mRNA capping. The Severe Acute Respiratory Syndrome coronavirus nsp16 protein is a S adenosylmethionine-dependent (nucleoside-2'-O)-methyltransferase only active in the presence of its activating partner nsp10. We report the nsp10/nsp16 complex structure at 2.0 A resolution, which shows nsp10 bound to nsp16 through a ~930 A2 surface area in nsp10. Functional assays identify key residues involved in nsp10/nsp16 association, and in RNA binding or catalysis, the latter likely through a SN2-like mechanism. We present two other crystal structures, the inhibitor Sinefungin bound in the S-adenosylmethionine binding pocket and the tighter complex nsp10(Y96F)/nsp16, providing the first structural insight into the regulation of RNA capping enzymes in +RNA viruses. PMID- 21637814 TI - Phospholipids trigger Cryptococcus neoformans capsular enlargement during interactions with amoebae and macrophages. AB - A remarkable aspect of the interaction of Cryptococcus neoformans with mammalian hosts is a consistent increase in capsule volume. Given that many aspects of the interaction of C. neoformans with macrophages are also observed with amoebae, we hypothesized that the capsule enlargement phenomenon also had a protozoan parallel. Incubation of C. neoformans with Acanthamoeba castellanii resulted in C. neoformans capsular enlargement. The phenomenon required contact between fungal and protozoan cells but did not require amoeba viability. Analysis of amoebae extracts showed that the likely stimuli for capsule enlargement were protozoan polar lipids. Extracts from macrophages and mammalian serum also triggered cryptococcal capsular enlargement. C. neoformans capsule enlargement required expression of fungal phospholipase B, but not phospholipase C. Purified phospholipids, in particular, phosphatidylcholine, and derived molecules triggered capsular enlargement with the subsequent formation of giant cells. These results implicate phospholipids as a trigger for both C. neoformans capsule enlargement in vivo and exopolysaccharide production. The observation that the incubation of C. neoformans with phospholipids led to the formation of giant cells provides the means to generate these enigmatic cells in vitro. Protozoan- or mammalian-derived polar lipids could represent a danger signal for C. neoformans that triggers capsular enlargement as a non-specific defense mechanism against potential predatory cells. Hence, phospholipids are the first host derived molecules identified to trigger capsular enlargement. The parallels apparent in the capsular response of C. neoformans to both amoebae and macrophages provide additional support for the notion that certain aspects of cryptococcal virulence emerged as a consequence of environmental interactions with other microorganisms such as protists. PMID- 21637815 TI - The merging of two dynasties--identification of an African cotton leaf curl disease-associated begomovirus with cotton in Pakistan. AB - Cotton leaf curl disease (CLCuD) is a severe disease of cotton that occurs in Africa and Pakistan/northwestern India. The disease is caused by begomoviruses in association with specific betasatellites that differ between Africa and Asia. During survey of symptomatic cotton in Sindh (southern Pakistan) Cotton leaf curl Gezira virus (CLCuGV), the begomovirus associated with CLCuD in Africa, was identified. However, the cognate African betasatellite (Cotton leaf curl Gezira betasatellite) was not found. Instead, two Asian betasatellites, the CLCuD associated Cotton leaf curl Multan betasatellite (CLCuMB) and Chilli leaf curl betasatellite (ChLCB) were identified. Inoculation of the experimental plant species Nicotiana benthamiana showed that CLCuGV was competent to maintain both CLCuMB and ChLCB. Interestingly, the enations typical of CLCuD were only induced by CLCuGV in the presence of CLCuMB. Also in infections involving both CLCuMB and ChLCB the enations typical of CLCuMB were less evident. This is the first time an African begomovirus has been identified on the Indian sub-continent, highlight the growing threat of begomoviruses and particularly the threat of CLCuD causing viruses to cotton cultivation in the rest of the world. PMID- 21637816 TI - The Coxiella burnetii Dot/Icm system delivers a unique repertoire of type IV effectors into host cells and is required for intracellular replication. AB - Coxiella burnetii, the causative agent of human Q fever, is an intracellular pathogen that replicates in an acidified vacuole derived from the host lysosomal network. This pathogen encodes a Dot/Icm type IV secretion system that delivers bacterial proteins called effectors to the host cytosol. To identify new effector proteins, the functionally analogous Legionella pneumophila Dot/Icm system was used in a genetic screen to identify fragments of C. burnetii genomic DNA that when fused to an adenylate cyclase reporter were capable of directing Dot/Icm dependent translocation of the fusion protein into mammalian host cells. This screen identified Dot/Icm effectors that were proteins unique to C. burnetii, having no overall sequence homology with L. pneumophila Dot/Icm effectors. A comparison of C. burnetii genome sequences from different isolates revealed diversity in the size and distribution of the genes encoding many of these effectors. Studies examining the localization and function of effectors in eukaryotic cells provided evidence that several of these proteins have an affinity for specific host organelles and can disrupt cellular functions. The identification of a transposon insertion mutation that disrupts the dot/icm locus was used to validate that this apparatus was essential for translocation of effectors. Importantly, this C. burnetii Dot/Icm-deficient mutant was found to be defective for intracellular replication. Thus, these data indicate that C. burnetii encodes a unique subset of bacterial effector proteins translocated into host cells by the Dot/Icm apparatus, and that the cumulative activities exerted by these effectors enables C. burnetii to successfully establish a niche inside mammalian cells that supports intracellular replication. PMID- 21637817 TI - OsSpo11-4, a rice homologue of the archaeal TopVIA protein, mediates double strand DNA cleavage and interacts with OsTopVIB. AB - DNA topoisomerase VI from Archaea, a heterotetrameric complex composed of two TopVIA and two TopVIB subunits, is involved in altering DNA topology during replication, transcription and chromosome segregation by catalyzing DNA strand transfer through transient double-strand breaks. The sequenced yeast and animal genomes encode only one homologue of the archaeal TopVIA subunit, namely Spo11, and no homologue of the archaeal TopVIB subunit. In yeast, Spo11 is essential for initiating meiotic recombination and this function appears conserved among other eukaryotes. In contrast to yeast and animals, studies in Arabidopsis and rice have identified three Spo11/TopVIA homologues and one TopVIB homologue in plants. Here, we further identified two novel Spo11/TopVIA homologues (named OsSpo11-4 and OsSpo11-5, respectively) that exist just in the monocot model plant Oryza sativa, indicating that at least five Spo11/TopVIA homologues are present in the rice genome. To reveal the biochemical function of the two novel Spo11/TopVIA homologues, we first examined the interactions among OsSpo11-1, OsSpo11-4, OsSpo11-5, and OsTopVIB by yeast two-hybrid assay. The results showed that OsSpo11-4 and OsTopVIB can self-interact strongly and among the 3 examined OsSpo11 proteins, only OsSpo11-4 interacted with OsTopVIB. Pull-down assay confirmed the interaction between OsSpo11-4 and OsTopVIB, which indicates that OsSpo11-4 may interact with OsTopVIB in vivo. Further in vitro enzymatic analysis revealed that among the above 4 proteins, only OsSpo11-4 exhibited double-strand DNA cleavage activity and its enzymatic activity appears dependent on Mg(2+) and independent of OsTopVIB, despite its interaction with OsTopVIB. We further analyzed the biological function of OsSpo11-4 by RNA interference and found that down-regulated expression of OsSpo11-4 led to defects in male meiosis, indicating OsSpo11-4 is required for meiosis. PMID- 21637818 TI - Fluid shear stress regulates the invasive potential of glioma cells via modulation of migratory activity and matrix metalloproteinase expression. AB - BACKGROUND: Glioma cells are exposed to elevated interstitial fluid flow during the onset of angiogenesis, at the tumor periphery while invading normal parenchyma, within white matter tracts, and during vascular normalization therapy. Glioma cell lines that have been exposed to fluid flow forces in vivo have much lower invasive potentials than in vitro cell motility assays without flow would indicate. METHODOLOGY/PRINCIPAL FINDINGS: A 3D Modified Boyden chamber (Darcy flow through collagen/cell suspension) model was designed to mimic the fluid dynamic microenvironment to study the effects of fluid shear stress on the migratory activity of glioma cells. Novel methods for gel compaction and isolation of chemotactic migration from flow stimulation were utilized for three glioma cell lines: U87, CNS-1, and U251. All physiologic levels of fluid shear stress suppressed the migratory activity of U87 and CNS-1 cell lines. U251 motility remained unaltered within the 3D interstitial flow model. Matrix Metalloproteinase (MMP) inhibition experiments and assays demonstrated that the glioma cells depended on MMP activity to invade, and suppression in motility correlated with downregulation of MMP-1 and MMP-2 levels. This was confirmed by RT-PCR and with the aid of MMP-1 and MMP-2 shRNA constructs. CONCLUSIONS/SIGNIFICANCE: Fluid shear stress in the tumor microenvironment may explain reduced glioma invasion through modulation of cell motility and MMP levels. The flow-induced migration trends were consistent with reported invasive potentials of implanted gliomas. The models developed for this study imply that flow-modulated motility involves mechanotransduction of fluid shear stress affecting MMP activation and expression. These models should be useful for the continued study of interstitial flow effects on processes that affect tumor progression. PMID- 21637819 TI - Stromal down-regulation of macrophage CD4/CCR5 expression and NF-kappaB activation mediates HIV-1 non-permissiveness in intestinal macrophages. AB - Tissue macrophages are derived exclusively from blood monocytes, which as monocyte-derived macrophages support HIV-1 replication. However, among human tissue macrophages only intestinal macrophages are non-permissive to HIV-1, suggesting that the unique microenvironment in human intestinal mucosa renders lamina propria macrophages non-permissive to HIV-1. We investigated this hypothesis using blood monocytes and intestinal extracellular matrix (stroma) conditioned media (S-CM) to model the exposure of newly recruited monocytes and resident macrophages to lamina propria stroma, where the cells take up residence in the intestinal mucosa. Exposure of monocytes to S-CM blocked up-regulation of CD4 and CCR5 expression during monocyte differentiation into macrophages and inhibited productive HIV-1 infection in differentiated macrophages. Importantly, exposure of monocyte-derived macrophages simultaneously to S-CM and HIV-1 also inhibited viral replication, and sorted CD4+ intestinal macrophages, a proportion of which expressed CCR5+, did not support HIV-1 replication, indicating that the non-permissiveness to HIV-1 was not due to reduced receptor expression alone. Consistent with this conclusion, S-CM also potently inhibited replication of HIV 1 pseudotyped with vesicular stomatitis virus glycoprotein, which provides CD4/CCR5-independent entry. Neutralization of TGF-beta in S-CM and recombinant TGF-beta studies showed that stromal TGF-beta inhibited macrophage nuclear translocation of NF-kappaB and HIV-1 replication. Thus, the profound inability of intestinal macrophages to support productive HIV-1 infection is likely the consequence of microenvironmental down-regulation of macrophage HIV-1 receptor/coreceptor expression and NF-kappaB activation. PMID- 21637820 TI - The cell surface proteome of human mesenchymal stromal cells. AB - BACKGROUND: Multipotent human mesenchymal stromal cells (hMSCs) are considered as promising biological tools for regenerative medicine. Their antibody-based isolation relies on the identification of reliable cell surface markers. METHODOLOGY/PRINCIPAL FINDINGS: To obtain a comprehensive view of the cell surface proteome of bone marrow-derived hMSCs, we have developed an analytical pipeline relying on cell surface biotinylation of intact cells using cell impermeable, cleavable sulfo-NHS-SS-biotin to enrich the plasma membrane proteins and mass spectrometry for identification with extremely high confidence. Among the 888 proteins identified, we found ~200 bona fide plasma membrane proteins including 33 cell adhesion molecules and 26 signaling receptors. In total 41 CD markers including 5 novel ones (CD97, CD112, CD239, CD276, and CD316) were identified. The CD markers are distributed homogenously within plastic-adherent hMSC populations and their expression is modulated during the process of adipogenesis or osteogenesis. Moreover, our in silico analysis revealed a significant difference between the cell surface proteome of hMSCs and that of human embryonic stem cells reported previously. CONCLUSIONS/SIGNIFICANCE: Collectively, our analytical methods not only provide a basis for further studies of mechanisms maintaining the multipotency of hMSCs within their niches and triggering their differentiation after signaling, but also a toolbox for a refined antibody-based identification of hMSC populations from different tissues and their isolation for therapeutic intervention. PMID- 21637821 TI - Proteomic analysis of honeybee (Apis mellifera L.) pupae head development. AB - The honeybee pupae development influences its future adult condition as well as honey and royal jelly productions. However, the molecular mechanism that regulates honeybee pupae head metamorphosis is still poorly understood. To further our understand of the associated molecular mechanism, we investigated the protein change of the honeybee pupae head at 5 time-points using 2-D electrophoresis, mass spectrometry, bioinformatics, quantitative real-time polymerase chain reaction and Western blot analysis. Accordingly, 58 protein spots altered their expression across the 5 time points (13-20 days), of which 36 proteins involved in the head organogenesis were upregulated during early stages (13-17 days). However, 22 proteins involved in regulating the pupae head neuron and gland development were upregulated at later developmental stages (19-20 days). Also, the functional enrichment analysis further suggests that proteins related to carbohydrate metabolism and energy production, development, cytoskeleton and protein folding were highly involved in the generation of organs and development of honeybee pupal head. Furthermore, the constructed protein interaction network predicted 33 proteins acting as key nodes of honeybee pupae head growth of which 9 and 4 proteins were validated at gene and protein levels, respectively. In this study, we uncovered potential protein species involved in the formation of honeybee pupae head development along with their specific temporal requirements. This first proteomic result allows deeper understanding of the proteome profile changes during honeybee pupae head development and provides important potential candidate proteins for future reverse genetic research on honeybee pupae head development to improve the performance of related organs. PMID- 21637822 TI - Detection of plant volatiles after leaf wounding and darkening by proton transfer reaction "time-of-flight" mass spectrometry (PTR-TOF). AB - Proton transfer reaction-time of flight (PTR-TOF) mass spectrometry was used to improve detection of biogenic volatiles organic compounds (BVOCs) induced by leaf wounding and darkening. PTR-TOF measurements unambiguously captured the kinetic of the large emissions of green leaf volatiles (GLVs) and acetaldehyde after wounding and darkening. GLVs emission correlated with the extent of wounding, thus confirming to be an excellent indicator of mechanical damage. Transient emissions of methanol, C5 compounds and isoprene from plant species that do not emit isoprene constitutively were also detected after wounding. In the strong isoprene-emitter Populus alba, light-dependent isoprene emission was sustained and even enhanced for hours after photosynthesis inhibition due to leaf cutting. Thus isoprene emission can uncouple from photosynthesis and may occur even after cutting leaves or branches, e.g., by agricultural practices or because of abiotic and biotic stresses. This observation may have important implications for assessments of isoprene sources and budget in the atmosphere, and consequences for tropospheric chemistry. PMID- 21637823 TI - Cancer induces cardiomyocyte remodeling and hypoinnervation in the left ventricle of the mouse heart. AB - Cancer is often associated with cachexia, cardiovascular symptoms and autonomic dysregulation. We tested whether extracardiac cancer directly affects the innervation of left ventricular myocardium. Mice injected with Lewis lung carcinoma cells (tumor group, TG) or PBS (control group, CG) were analyzed after 21 days. Cardiac function (echocardiography), serum levels of TNF-alpha and Il-6 (ELISA), structural alterations of cardiomyocytes and their innervation (design based stereology) and levels of innervation-related mRNA (quantitative RT-PCR) were analysed. The groups did not differ in various functional parameters. Serum levels of TNF-alpha and Il-6 were elevated in TG. The total length of axons in the left ventricle was reduced. The number of dense core vesicles per axon profile was reduced. Decreased myofibrillar volume, increased sarcoplasmic volume and increased volume of lipid droplets were indicative of metabolic alterations of TG cardiomyocytes. In the heart, the mRNA level of nerve growth factor was reduced whereas that of beta1-adrenergic receptor was unchanged in TG. In the stellate ganglion of TG, mRNA levels of nerve growth factor and neuropeptide Y were decreased and that of tyrosine hydroxylase was increased. In summary, cancer induces a systemic pro-inflammatory state, a significant reduction in myocardial innervation and a catabolic phenotype of cardiomyocytes in the mouse. Reduced expression of nerve growth factor may account for the reduced myocardial innervation. PMID- 21637824 TI - Hybrid titanium/biodegradable polymer implants with an hierarchical pore structure as a means to control selective cell movement. AB - In order to improve implant success rate, it is important to enhance their responsiveness to the prevailing conditions following implantation. Uncontrolled movement of inflammatory cells and fibroblasts is one of these in vivo problems and the porosity properties of the implant have a strong effect on these. Here, we describe a hybrid system composed of a macroporous titanium structure filled with a microporous biodegradable polymer. This polymer matrix has a distinct porosity gradient to accommodate different cell types (fibroblasts and epithelial cells). The main clinical application of this system will be the prevention of restenosis due to excessive fibroblast migration and proliferation in the case of tracheal implants. METHODOLOGY/PRINCIPAL FINDINGS: A microbead-based titanium template was filled with a porous Poly (L-lactic acid) (PLLA) body by freeze extraction method. A distinct porosity difference was obtained between the inner and outer surfaces of the implant as characterized by image analysis and Mercury porosimetry (9.8+/-2.2 um vs. 36.7+/-11.4 um, p<=0.05). On top, a thin PLLA film was added to optimize the growth of epithelial cells, which was confirmed by using human respiratory epithelial cells. To check the control of fibroblast movement, PKH26 labeled fibroblasts were seeded onto Titanium and Titanium/PLLA implants. The cell movement was quantified by confocal microscopy: in one week cells moved deeper in Ti samples compared to Ti/PLLA. CONCLUSIONS: In vitro experiments showed that this new implant is effective for guiding different kind of cells it will contact upon implantation. Overall, this system would enable spatial and temporal control over cell migration by a gradient ranging from macroporosity to nanoporosity within a tracheal implant. Moreover, mechanical properties will be dependent mainly on the titanium frame. This will make it possible to create a polymeric environment which is suitable for cells without the need to meet mechanical requirements with the polymeric structure. PMID- 21637826 TI - Symbiodinium genotypic and environmental controls on lipids in reef building corals. AB - BACKGROUND: Lipids in reef building corals can be divided into two classes; non polar storage lipids, e.g. wax esters and triglycerides, and polar structural lipids, e.g. phospholipids and cholesterol. Differences among algal endosymbiont types are known to have important influences on processes including growth and the photobiology of scleractinian corals yet very little is known about the role of symbiont types on lipid energy reserves. METHODOLOGY/PRINCIPAL FINDINGS: The ratio of storage lipid and structural lipid fractions of Scott Reef corals were determined by thin layer chromatography. The lipid fraction ratio varied with depth and depended on symbiont type harboured by two corals (Seriatopora hystrix and Pachyseris speciosa). S. hystrix colonies associated with Symbiodinium C1 or C1/C# at deep depths (>23 m) had lower lipid fraction ratios (i.e. approximately equal parts of storage and structural lipids) than those with Symbiodinium D1 in shallow depths (<23 m), which had higher lipid fraction ratios (i.e. approximately double amounts of storage relative to structural lipid). Further, there was a non-linear relationship between the lipid fraction ratio and depth for S. hystrix with a modal peak at ~23 m coinciding with the same depth as the shift from clade D to C types. In contrast, the proportional relationship between the lipid fraction ratio and depth for P. speciosa, which exhibited high specificity for Symbiodinium C3 like across the depth gradient, was indicative of greater amounts of storage lipids contained in the deep colonies. CONCLUSIONS/SIGNIFICANCE: This study has demonstrated that Symbiodinium exert significant controls over the quality of coral energy reserves over a large-scale depth gradient. We conclude that the competitive advantages and metabolic costs that arise from flexible associations with divergent symbiont types are offset by energetic trade-offs for the coral host. PMID- 21637825 TI - NKX2-3 transcriptional regulation of endothelin-1 and VEGF signaling in human intestinal microvascular endothelial cells. AB - BACKGROUND: NKX2-3 is associated with inflammatory bowel disease (IBD). NKX2-3 is expressed in microvascular endothelial cells and the muscularis mucosa of the gastrointestinal tract. Human intestinal microvascular endothelial cells (HIMECs) are actively involved in the pathogenesis of IBD and IBD-associated microvascular dysfunction. To understand the cellular function of NKX2-3 and its potential role underlying IBD pathogenesis, we investigated the genes regulated by NKX2-3 in HIMEC using cDNA microarray. METHODOLOGY/PRINCIPAL FINDINGS: NKX2-3 expression was suppressed by shRNA in two HIMEC lines and gene expression was profiled by cDNA microarray. Pathway Analysis was used to identify gene networks according to biological functions and associated pathways. Validation of microarray and genes expression in intestinal tissues was assessed by RT-PCR. NKX2-3 regulated genes are involved in immune and inflammatory response, cell proliferation and growth, metabolic process, and angiogenesis. Several inflammation and angiogenesis related signaling pathways that play important roles in IBD were regulated by NKX2-3, including endothelin-1 and VEGF-PI3K/AKT-eNOS. Expression levels of NKX2 3, VEGFA, PI3K, AKT, and eNOS are increased in intestinal tissues from IBD patients and expression levels of EDN1 are decreased in intestinal tissues from IBD patients. These results demonstrated the important roles of NKX2-3, VEGF, PI3K, AKT, eNOS, and EDN1 in IBD pathogenesis. Correlation analysis showed a positive correlation between mRNA expression of NKX2-3 and VEGFA and a negative correlation between mRNA expression of NKX2-3 and EDN1 in intestinal tissues from IBD patients. CONCLUSION/RELEVANCE: NKX2-3 may play an important role in IBD pathogenesis by regulating endothelin-1 and VEGF signaling in HIMECs. PMID- 21637827 TI - Cross-mapping events in miRNAs reveal potential miRNA-mimics and evolutionary implications. AB - MicroRNAs (miRNAs) have important roles in various biological processes. miRNA cross-mapping is a prevalent phenomenon where miRNA sequence originating from one genomic region is mapped to another location. To have a better understanding of this phenomenon in the human genome, we performed a detailed analysis in this paper using public miRNA high-throughput sequencing data and all known human miRNAs. We observed widespread cross-mapping events between miRNA precursors (pre miRNAs), other non-coding RNAs (ncRNAs) and the opposite strands of pre-miRNAs by analyzing the high-throughput sequencing data. Computational analysis on all known human miRNAs also confirmed that many of them could be involved in cross mapping events. The processing or decay of both ncRNAs and pre-miRNA opposite strand transcripts may contribute to miRNA enrichment, although some might be miRNA-mimics due to miRNA mis-annotation. Comparing to canonical miRNAs, miRNAs involved in cross-mapping events between pre-miRNAs and other ncRNAs normally had shorter lengths (17-19 nt), lower prediction scores and were classified as pseudo miRNA precursors. Notably, 4.9% of all human miRNAs could be accurately mapped to the opposite strands of pre-miRNAs, which showed that both strands of the same genomic region had the potential to produce mature miRNAs and simultaneously implied some potential miRNA precursors. We proposed that the cross-mapping events are more complex than we previously thought. Sequence similarity between other ncRNAs and pre-miRNAs and the specific stem-loop structures of pre-miRNAs may provide evolutionary implications. PMID- 21637829 TI - Failure tolerance of motif structure in biological networks. AB - Complex networks serve as generic models for many biological systems that have been shown to share a number of common structural properties such as power-law degree distribution and small-worldness. Real-world networks are composed of building blocks called motifs that are indeed specific subgraphs of (usually) small number of nodes. Network motifs are important in the functionality of complex networks, and the role of some motifs such as feed-forward loop in many biological networks has been heavily studied. On the other hand, many biological networks have shown some degrees of robustness in terms of their efficiency and connectedness against failures in their components. In this paper we investigated how random and systematic failures in the edges of biological networks influenced their motif structure. We considered two biological networks, namely, protein structure network and human brain functional network. Furthermore, we considered random failures as well as systematic failures based on different strategies for choosing candidate edges for removal. Failure in the edges tipping to high degree nodes had the most destructive role in the motif structure of the networks by decreasing their significance level, while removing edges that were connected to nodes with high values of betweenness centrality had the least effect on the significance profiles. In some cases, the latter caused increase in the significance levels of the motifs. PMID- 21637828 TI - Deep sequencing reveals novel microRNAs and regulation of microRNA expression during cell senescence. AB - In cell senescence, cultured cells cease proliferating and acquire aberrant gene expression patterns. MicroRNAs (miRNAs) modulate gene expression through translational repression or mRNA degradation and have been implicated in senescence. We used deep sequencing to carry out a comprehensive survey of miRNA expression and involvement in cell senescence. Informatic analysis of small RNA sequence datasets from young and senescent IMR90 human fibroblasts identifies many miRNAs that are regulated (either up or down) with cell senescence. Comparison with mRNA expression profiles reveals potential mRNA targets of these senescence-regulated miRNAs. The target mRNAs are enriched for genes involved in biological processes associated with cell senescence. This result greatly extends existing information on the role of miRNAs in cell senescence and is consistent with miRNAs having a causal role in the process. PMID- 21637830 TI - ECAT11/L1td1 is enriched in ESCs and rapidly activated during iPSC generation, but it is dispensable for the maintenance and induction of pluripotency. AB - The principal factors that lead to proliferation and pluripotency in embryonic stem cells (ESCs) have been vigorously investigated. However, the global network of factors and their full signaling cascade is still unclear. In this study, we found that ECAT11 (L1td1) is one of the ESC-associated transcripts harboring a truncated fragment of ORF-1, a component of the L1 retrotransposable element. We generated an ECAT11 knock-in mouse by replacing its coding region with green fluorescent protein. In the early stage of development, the fluorescence was observed at the inner cell mass of blastocysts and epiblasts. Despite this specific expression, ECAT11-null mice grow normally and are fertile. In addition, ECAT11 was dispensable for both the proliferation and pluripotency of ESCs.We found rapid and robust activation of ECAT11 in fibroblasts after the forced expression of transcription factors that can give rise pluripotency in somatic cells. However, iPS cells could be established from ECAT11-null fibroblasts. Our data demonstrate the dispensability of ECAT11/L1td1 in pluripotency, despite its specific expression. PMID- 21637831 TI - Altered cerebellar-cerebral functional connectivity in geriatric depression. AB - Although volumetric and activation changes in the cerebellum have frequently been reported in studies on major depression, its role in the neural mechanism of depression remains unclear. To understand how the cerebellum may relate to affective and cognitive dysfunction in depression, we investigated the resting state functional connectivity between cerebellar regions and the cerebral cortex in samples of patients with geriatric depression (n = 11) and healthy controls (n = 18). Seed-based connectivity analyses were conducted using seeds from cerebellum regions previously identified as being involved in the executive, default-mode, affective-limbic, and motor networks. The results revealed that, compared with controls, individuals with depression show reduced functional connectivity between several cerebellum seed regions, specifically those in the executive and affective-limbic networks with the ventromedial prefrontal cortex (vmPFC) and increased functional connectivity between the motor-related cerebellum seed regions with the putamen and motor cortex. We further investigated whether the altered functional connectivity in depressed patients was associated with cognitive function and severity of depression. A positive correlation was found between the Crus II-vmPFC connectivity and performance on the Hopkins Verbal Learning Test-Revised delayed memory recall. Additionally, the vermis-posterior cinglate cortex (PCC) connectivity was positively correlated with depression severity. Our results suggest that cerebellum-vmPFC coupling may be related to cognitive function whereas cerebellum-PCC coupling may be related to emotion processing in geriatric depression. PMID- 21637832 TI - Comparative analyses by sequencing of transcriptomes during skeletal muscle development between pig breeds differing in muscle growth rate and fatness. AB - Understanding the dynamics of muscle transcriptome during development and between breeds differing in muscle growth is necessary to uncover the complex mechanism underlying muscle development. Herein, we present the first transcriptome-wide longissimus dorsi muscle development research concerning Lantang (LT, obese) and Landrace (LR, lean) pig breeds during 10 time-points from 35 days-post-coitus (dpc) to 180 days-post-natum (dpn) using Solexa/Illumina's Genome Analyzer. The data demonstrated that myogenesis was almost completed before 77 dpc, but the muscle phenotypes were still changed from 77 dpc to 28 dpn. Comparative analysis of the two breeds suggested that myogenesis started earlier but progressed more slowly in LT than in LR, the stages ranging from 49 dpc to 77 dpc are critical for formation of different muscle phenotypes. 595 differentially expressed myogenesis genes were identified, and their roles in myogenesis were discussed. Furthermore, GSK3B, IKBKB, ACVR1, ITGA and STMN1 might contribute to later myogenesis and more muscle fibers in LR than LT. Some myogenesis inhibitors (ID1, ID2, CABIN1, MSTN, SMAD4, CTNNA1, NOTCH2, GPC3 and HMOX1) were higher expressed in LT than in LR, which might contribute to more slow muscle differentiation in LT than in LR. We also identified several genes which might contribute to intramuscular adipose differentiation. Most important, we further proposed a novel model in which MyoD and MEF2A controls the balance between intramuscular adipogenesis and myogenesis by regulating CEBP family; Myf5 and MEF2C are essential during the whole myogenesis process while MEF2D affects muscle growth and maturation. The MRFs and MEF2 families are also critical for the phenotypic differences between the two pig breeds. Overall, this study contributes to elucidating the mechanism underlying muscle development, which could provide valuable information for pig meat quality improvement. The raw data have been submitted to Gene Expression Omnibus (GEO) under series GSE25406. PMID- 21637833 TI - Parent-completed developmental screening in premature children: a valid tool for follow-up programs. AB - Our goals were to (1) validate the parental Ages and Stages Questionnaires (ASQ) as a screening tool for psychomotor development among a cohort of ex-premature infants reaching 2 years, and (2) analyse the influence of parental socio economic status and maternal education on the efficacy of the questionnaire. A regional population of 703 very preterm infants (<35 weeks gestational age) born between 2003 and 2006 were evaluated at 2 years by their parents who completed the ASQ, by a pediatric clinical examination, and by the revised Brunet Lezine psychometric test with establishment of a DQ score. Detailed information regarding parental socio-economic status was available for 419 infants. At 2 years corrected age, 630 infants (89.6%) had an optimal neuromotor examination. Overall ASQ scores for predicting a DQ score <=85 produced an area under the receiver operator curve value of 0.85 (95% Confidence Interval:0.82-0.87). An ASQ cut-off score of <=220 had optimal discriminatory power for identifying a DQ score <=85 with a sensitivity of 0.85 (95%CI:0.75-0.91), a specificity of 0.72 (95%CI:0.69-0.75), a positive likelihood ratio of 3, and a negative likelihood ratio of 0.21. The median value for ASQ was not significantly associated with socio-economic level or maternal education. ASQ is an easy and reliable tool regardless of the socio-economic status of the family to predict normal neurologic outcome in ex-premature infants at 2 years of age. ASQ may be beneficial with a low-cost impact to some follow-up programs, and helps to establish a genuine sense of parental involvement. PMID- 21637834 TI - Drosophila eggshell production: identification of new genes and coordination by Pxt. AB - Drosophila ovarian follicles complete development using a spatially and temporally controlled maturation process in which they resume meiosis and secrete a multi-layered, protective eggshell before undergoing arrest and/or ovulation. Microarray analysis revealed more than 150 genes that are expressed in a stage specific manner during the last 24 hours of follicle development. These include all 30 previously known eggshell genes, as well as 19 new candidate chorion genes and 100 other genes likely to participate in maturation. Mutations in pxt, encoding a putative Drosophila cyclooxygenase, cause many transcripts to begin expression prematurely, and are associated with eggshell defects. Somatic activity of Pxt is required, as RNAi knockdown of pxt in the follicle cells recapitulates both the temporal expression and eggshell defects. One of the temporally regulated genes, cyp18a1, which encodes a cytochromome P450 protein mediating ecdysone turnover, is downregulated in pxt mutant follicles, and cyp18a1 mutation itself alters eggshell gene expression. These studies further define the molecular program of Drosophila follicle maturation and support the idea that it is coordinated by lipid and steroid hormonal signals. PMID- 21637835 TI - Herpes simplex virus type 2, genital ulcers and HIV-1 disease progression in postpartum women. AB - BACKGROUND: Co-infection with herpes simplex virus type 2 (HSV-2) has been associated with increased HIV-1 RNA levels and immune activation, two predictors of HIV-1 progression. The impact of HSV-2 on clinical outcomes among HIV-1 infected pregnant women is unclear. METHODS: HIV-1 infected pregnant women in Nairobi were enrolled antenatally and HSV-2 serology was obtained. HIV-1 RNA and CD4 count were serially measured for 12-24 months postpartum. Survival analysis using endpoints of death, opportunistic infection (OI), and CD4<200 cells uL, and linear mixed models estimating rate of change of HIV-1 RNA and CD4, were used to determine associations between HSV-2 serostatus and HIV-1 progression. RESULTS: Among 296 women, 254 (86%) were HSV-2-seropositive. Only 30 (10%) women had prior or current genital ulcer disease (GUD); median baseline CD4 count was 422 cells uL. Adjusting for baseline CD4, women with GUD were significantly more likely to have incident OIs (adjusted hazard ratio (aHR) 2.79, 95% CI: 1.33-5.85), and there was a trend for association between HSV-2-seropositivity and incident OIs (aHR 3.83, 95% CI: 0.93-15.83). Rate of change in CD4 count and HIV-1 RNA did not differ by HSV-2 status or GUD, despite a trend toward higher baseline HIV-1 RNA in HSV-2-seropositive women (4.73 log10 copies/ml vs. 4.47 log10 copies/ml, P = 0.07). CONCLUSIONS: HSV-2 was highly prevalent and pregnant HIV-1 infected women with GUD were significantly more likely to have incident OIs than women without GUD, suggesting that clinically evident HSV-2 is a more important predictor of HIV-1 disease progression than asymptomatic HSV-2. PMID- 21637836 TI - An appraisal of the maternal mortality decline in Nepal. AB - BACKGROUND: A decline in the national maternal mortality ratio in Nepal has been observed from surveys conducted between 1996 and 2008. This paper aims to assess the plausibility of the decline and to identify drivers of change. METHODS: National and sub-national trends in mortality data were investigated using existing demographic and health surveys and maternal mortality and morbidity surveys. Potential drivers of the variation in maternal mortality between districts were identified by regressing district-level indicators from the Nepal demographic health surveys against maternal mortality estimates. RESULTS: A statistically significant decline of the maternal mortality ratio from 539 maternal deaths to 281 per 100,000 (95% CI 91,507) live births between 1993 and 2003 was demonstrated. The sub-national changes are of similar magnitude and direction to those observed nationally, and in the terai region (plains) the differences are statistically significant with a reduction of 361 per 100,000 live births (95% CI 36,686) during the same time period. The reduction in fertility, changes in education and wealth, improvements in components of the human development index, gender empowerment and anaemia each explained more than 10% of the district variation in maternal mortality. A number of limitations in each of the data sources used were identified. Of these, the most important relate to the underestimation of numbers of deaths. CONCLUSION: It is likely that there has been a decline in Nepal's maternal mortality since 1993. This is good news for the country's sustained commitments in this area. Conclusions on the magnitude, pattern of the change and drivers of the decline are constrained by lack of data. We recommend close tracking of maternal mortality and its determinants in Nepal, attention to the communication of future estimates, and various options for bridging data gaps. PMID- 21637837 TI - A set of 100 chloroplast DNA primer pairs to study population genetics and phylogeny in monocotyledons. AB - Chloroplast DNA sequences are of great interest for population genetics and phylogenetic studies. However, only a small set of markers are commonly used. Most of them have been designed for amplification in a large range of Angiosperms and are located in the Large Single Copy (LSC). Here we developed a new set of 100 primer pairs optimized for amplification in Monocotyledons. Primer pairs amplify coding (exon) and non-coding regions (intron and intergenic spacer). They span the different chloroplast regions: 72 are located in the LSC, 13 in the Small Single Copy (SSC) and 15 in the Inverted Repeat region (IR). Amplification and sequencing were tested in 13 species of Monocotyledons: Dioscorea abyssinica, D. praehensilis, D. rotundata, D. dumetorum, D. bulbifera, Trichopus sempervirens (Dioscoreaceae), Phoenix canariensis, P. dactylifera, Astrocaryum scopatum, A. murumuru, Ceroxylon echinulatum (Arecaceae), Digitaria excilis and Pennisetum glaucum (Poaceae). The diversity found in Dioscorea, Digitaria and Pennisetum mainly corresponded to Single Nucleotide Polymorphism (SNP) while the diversity found in Arecaceae also comprises Variable Number Tandem Repeat (VNTR). We observed that the most variable loci (rps15-ycf1, rpl32-ccsA, ndhF-rpl32, ndhG ndhI and ccsA) are located in the SSC. Through the analysis of the genetic structure of a wild-cultivated species complex in Dioscorea, we demonstrated that this new set of primers is of great interest for population genetics and we anticipate that it will also be useful for phylogeny and bar-coding studies. PMID- 21637838 TI - Identification of novel targets of CSL-dependent Notch signaling in hematopoiesis. AB - Somatic activating mutations in the Notch1 receptor result in the overexpression of activated Notch1, which can be tumorigenic. The goal of this study is to understand the molecular mechanisms underlying the phenotypic changes caused by the overexpression of ligand independent Notch 1 by using a tetracycline inducible promoter in an in vitro embryonic stem (ES) cells/OP9 stromal cells coculture system, recapitulating normal hematopoiesis. First, an in silico analysis of the promoters of Notch regulated genes (previously determined by microarray analysis) revealed that the motifs recognized by regulatory proteins known to mediate hematopoiesis were overrepresented. Notch 1 does not bind DNA but instead binds the CSL transcription factor to regulate gene expression. The in silico analysis also showed that there were putative CSL binding sites observed in the promoters of 28 out of 148 genes. A custom ChIP-chip array was used to assess the occupancy of CSL in the promoter regions of the Notch1 regulated genes in vivo and showed that 61 genes were bound by activated Notch responsive CSL. Then, comprehensive mapping of the CSL binding sites genome-wide using ChIP-seq analysis revealed that over 10,000 genes were bound within 10 kb of the TSS (transcription start site). The majority of the targets discovered by ChIP-seq belong to pathways that have been shown by others to crosstalk with Notch signaling. Finally, 83 miRNAs were significantly differentially expressed by greater than 1.5-fold during the course of in vitro hematopoiesis. Thirty one miRNA were up-regulated and fifty two were down-regulated. Overexpression of Notch1 altered this pattern of expression of microRNA: six miRNAs were up regulated and four were down regulated as a result of activated Notch1 overexpression during the course of hematopoiesis. Time course analysis of hematopoietic development revealed that cells with Notch 1 overexpression mimic miRNA expression of cells in a less mature stage, which is consistent with our previous biological characterization. PMID- 21637839 TI - Litter size variation in hypothalamic gene expression determines adult metabolic phenotype in Brandt's voles (Lasiopodomys brandtii). AB - BACKGROUND: Early postnatal environments may have long-term and potentially irreversible consequences on hypothalamic neurons involved in energy homeostasis. Litter size is an important life history trait and negatively correlated with milk intake in small mammals, and thus has been regarded as a naturally varying feature of the early developmental environment. Here we investigated the long term effects of litter size on metabolic phenotype and hypothalamic neuropeptide mRNA expression involved in the regulation of energy homeostasis, using the offspring reared from large (10-12) and small (3-4) litter sizes, of Brandt's voles (Lasiopodomys brandtii), a rodent species from Inner Mongolia grassland in China. METHODOLOGY/PRINCIPAL FINDINGS: Hypothalamic leptin signaling and neuropeptides were measured by Real-Time PCR. We showed that offspring reared from small litters were heavier at weaning and also in adulthood than offspring from large litters, accompanied by increased food intake during development. There were no significant differences in serum leptin levels or leptin receptor (OB-Rb) mRNA in the hypothalamus at weaning or in adulthood, however, hypothalamic suppressor of cytokine signaling 3 (SOCS3) mRNA in adulthood increased in small litters compared to that in large litters. As a result, the agouti-related peptide (AgRP) mRNA increased in the offspring from small litters. CONCLUSIONS/SIGNIFICANCE: These findings support our hypothesis that natural litter size has a permanent effect on offspring metabolic phenotype and hypothalamic neuropeptide expression, and suggest central leptin resistance and the resultant increase in AgRP expression may be a fundamental mechanism underlying hyperphagia and the increased risk of overweight in pups of small litters. Thus, we conclude that litter size may be an important and central determinant of metabolic fitness in adulthood. PMID- 21637840 TI - Quantitative proteomic and interaction network analysis of cisplatin resistance in HeLa cells. AB - Cisplatin along with other platinum based drugs are some of the most widely used chemotherapeutic agents. However drug resistance is a major problem for the successful chemotherapeutic treatment of cancer. Current evidence suggests that drug resistance is a multifactorial problem due to changes in the expression levels and activity of a wide number of proteins. A majority of the studies to date have quantified mRNA levels between drug resistant and drug sensitive cell lines. Unfortunately mRNA levels do not always correlate with protein expression levels due to post-transcriptional changes in protein abundance. Therefore global quantitative proteomics screens are needed to identify the protein targets that are differentially expressed in drug resistant cell lines. Here we employ a quantitative proteomics technique using stable isotope labeling with amino acids in cell culture (SILAC) coupled with mass spectrometry to quantify changes in protein levels between cisplatin resistant (HeLa/CDDP) and sensitive HeLa cells in an unbiased fashion. A total of 856 proteins were identified and quantified, with 374 displaying significantly altered expression levels between the cell lines. Expression level data was then integrated with a network of protein protein interactions, and biological pathways to obtain a systems level view of proteome changes which occur with cisplatin resistance. Several of these proteins have been previously implicated in resistance towards platinum-based and other drugs, while many represent new potential markers or therapeutic targets. PMID- 21637841 TI - Inhibition of bacterial conjugation by phage M13 and its protein g3p: quantitative analysis and model. AB - Conjugation is the main mode of horizontal gene transfer that spreads antibiotic resistance among bacteria. Strategies for inhibiting conjugation may be useful for preserving the effectiveness of antibiotics and preventing the emergence of bacterial strains with multiple resistances. Filamentous bacteriophages were first observed to inhibit conjugation several decades ago. Here we investigate the mechanism of inhibition and find that the primary effect on conjugation is occlusion of the conjugative pilus by phage particles. This interaction is mediated primarily by phage coat protein g3p, and exogenous addition of the soluble fragment of g3p inhibited conjugation at low nanomolar concentrations. Our data are quantitatively consistent with a simple model in which association between the pili and phage particles or g3p prevents transmission of an F plasmid encoding tetracycline resistance. We also observe a decrease in the donor ability of infected cells, which is quantitatively consistent with a reduction in pili elaboration. Since many antibiotic-resistance factors confer susceptibility to phage infection through expression of conjugative pili (the receptor for filamentous phage), these results suggest that phage may be a source of soluble proteins that slow the spread of antibiotic resistance genes. PMID- 21637842 TI - Search for limiting factors in the RNAi pathway in silkmoth tissues and the Bm5 cell line: the RNA-binding proteins R2D2 and Translin. AB - RNA interference (RNAi), an RNA-dependent gene silencing process that is initiated by double-stranded RNA (dsRNA) molecules, has been applied with variable success in lepidopteran insects, in contrast to the high efficiency achieved in the coleopteran Tribolium castaneum. To gain insight into the factors that determine the efficiency of RNAi, a survey was carried out to check the expression of factors that constitute the machinery of the small interfering RNA (siRNA) and microRNA (miRNA) pathways in different tissues and stages of the silkmoth, Bombyx mori. It was found that the dsRNA-binding protein R2D2, an essential component in the siRNA pathway in Drosophila, was expressed at minimal levels in silkmoth tissues. The silkmoth-derived Bm5 cell line was also deficient in expression of mRNA encoding full-length BmTranslin, an RNA-binding factor that has been shown to stimulate the efficiency of RNAi. However, despite the lack of expression of the RNA-binding proteins, silencing of a luciferase reporter gene was observed by co-transfection of luc dsRNA using a lipophilic reagent. In contrast, gene silencing was not detected when the cells were soaked in culture medium supplemented with dsRNA. The introduction of an expression construct for Tribolium R2D2 (TcR2D2) did not influence the potency of luc dsRNA to silence the luciferase reporter. Immunostaining experiments further showed that both TcR2D2 and BmTranslin accumulated at defined locations within the cytoplasm of transfected cells. Our results offer a first evaluation of the expression of the RNAi machinery in silkmoth tissues and Bm5 cells and provide evidence for a functional RNAi response to intracellular dsRNA in the absence of R2D2 and Translin. The failure of TcR2D2 to stimulate the intracellular RNAi pathway in Bombyx cells is discussed. PMID- 21637843 TI - Phosphoproteomics identifies oncogenic Ras signaling targets and their involvement in lung adenocarcinomas. AB - BACKGROUND: Ras is frequently mutated in a variety of human cancers, including lung cancer, leading to constitutive activation of MAPK signaling. Despite decades of research focused on the Ras oncogene, Ras-targeted phosphorylation events and signaling pathways have not been described on a proteome-wide scale. METHODOLOGY/PRINCIPAL FINDINGS: By functional phosphoproteomics, we studied the molecular mechanics of oncogenic Ras signaling using a pathway-based approach. We identified Ras-regulated phosphorylation events (n = 77) using label-free comparative proteomics analysis of immortalized human bronchial epithelial cells with and without the expression of oncogenic Ras. Many were newly identified as potential targets of the Ras signaling pathway. A majority (~60%) of the Ras targeted events consisted of a [pSer/Thr]-Pro motif, indicating the involvement of proline-directed kinases. By integrating the phosphorylated signatures into the Pathway Interaction Database, we further inferred Ras-regulated pathways, including MAPK signaling and other novel cascades, in governing diverse functions such as gene expression, apoptosis, cell growth, and RNA processing. Comparisons of Ras-regulated phosphorylation events, pathways, and related kinases in lung cancer-derived cells supported a role of oncogenic Ras signaling in lung adenocarcinoma A549 and H322 cells, but not in large cell carcinoma H1299 cells. CONCLUSIONS/SIGNIFICANCE: This study reveals phosphorylation events, signaling networks, and molecular functions that are regulated by oncogenic Ras. The results observed in this study may aid to extend our knowledge on Ras signaling in lung cancer. PMID- 21637844 TI - Directed neural differentiation of mouse embryonic stem cells is a sensitive system for the identification of novel Hox gene effectors. AB - The evolutionarily conserved Hox family of homeodomain transcription factors plays fundamental roles in regulating cell specification along the anterior posterior axis during development of all bilaterian animals by controlling cell fate choices in a highly localized, extracellular signal and cell context dependent manner. Some studies have established downstream target genes in specific systems but their identification is insufficient to explain either the ability of Hox genes to direct homeotic transformations or the breadth of their patterning potential. To begin delineating Hox gene function in neural development we used a mouse ES cell based system that combines efficient neural differentiation with inducible Hoxb1 expression. Gene expression profiling suggested that Hoxb1 acted as both activator and repressor in the short term but predominantly as a repressor in the long run. Activated and repressed genes segregated in distinct processes suggesting that, in the context examined, Hoxb1 blocked differentiation while activating genes related to early developmental processes, wnt and cell surface receptor linked signal transduction and cell-to cell communication. To further elucidate aspects of Hoxb1 function we used loss and gain of function approaches in the mouse and chick embryos. We show that Hoxb1 acts as an activator to establish the full expression domain of CRABPI and II in rhombomere 4 and as a repressor to restrict expression of Lhx5 and Lhx9. Thus the Hoxb1 patterning activity includes the regulation of the cellular response to retinoic acid and the delay of the expression of genes that commit cells to neural differentiation. The results of this study show that ES neural differentiation and inducible Hox gene expression can be used as a sensitive model system to systematically identify Hox novel target genes, delineate their interactions with signaling pathways in dictating cell fate and define the extent of functional overlap among different Hox genes. PMID- 21637845 TI - Diversity of SCCmec elements in methicillin-resistant coagulase-negative staphylococci clinical isolates. AB - BACKGROUND: Methicillin-resistant coagulase-negative staphylococci (MR-CoNS) are opportunistic pathogens and serve as a large reservoir of staphylococcal cassette chromosome mec (SCCmec). Characterization of SCCmec in MR-CoNS can generate useful information on the mobilization and evolution of this element. METHODOLOGY/PRINCIPAL FINDINGS: Non-repetitive MR-CoNS clinical isolates (n = 84; 39 S. epidermidis, 19 S. haemolyticus, 9 S. hominis, 6 S. capitis, 4 S. warneri, 2 S. cohnii, 2 S. saprophyticus, 1 S. kloosii, 1 S. simulans and 1 S. massiliensis) were collected. All isolates could grow on plates with 4 mg/L cefoxitin and all had mecA as detected by PCR. Strain typing using RAPD and ERIC PCR revealed that almost all isolates were of different strains. SCCmec typing was performed using multiplex PCR published previously. For isolates in which SCCmec could not be typed, the mec complex classes were determined by additional PCR and the ccr genes were amplified with published or newly-designed primers and then sequenced. SCCmec types were assigned for 63 isolates by multiplex PCR and were assigned for 14 other isolates by PCR targeting mec and ccr. Among 77 isolates with determined SCCmec types, 54 had a single type, including type III (n = 19), IV (n = 14), V (n = 10), II (n = 2), I (n = 1), VIII (n = 1) and five unnamed types (n = 7), while 23 isolates had two types, III+V (n = 12), II+V (n = 8), II+IV (n = 2) or IV+V (n = 1). The five unnamed types were assigned UT1 (class A mec, ccrA1/ccrB4), UT2 (class C1 mec, ccrA4/ccrB4), UT3 (class A mec, ccrA5/ccrB3), UT4 (class C2 mec, ccrA2/ccrB2 plus ccrC1) and UT5 (class A mec, ccrA1/ccrB1 plus ccrC1). CONCLUSIONS/SIGNIFICANCE: SCCmec types III, IV and V were prevalent in MR-CoNS and many isolates could harbor more than one type. Several new types of SCCmec were identified, highlighting the great genetic diversity and the need of developing classification schemes for SCCmec in MR CoNS. PMID- 21637846 TI - Heterologous expression of Alteromonas macleodii and Thiocapsa roseopersicina [NiFe] hydrogenases in Synechococcus elongatus. AB - Oxygen-tolerant [NiFe] hydrogenases may be used in future photobiological hydrogen production systems once the enzymes can be heterologously expressed in host organisms of interest. To achieve heterologous expression of [NiFe] hydrogenases in cyanobacteria, the two hydrogenase structural genes from Alteromonas macleodii Deep ecotype (AltDE), hynS and hynL, along with the surrounding genes in the gene operon of HynSL were cloned in a vector with an IPTG-inducible promoter and introduced into Synechococcus elongatus PCC7942. The hydrogenase protein was expressed at the correct size upon induction with IPTG. The heterologously-expressed HynSL hydrogenase was active when tested by in vitro H(2) evolution assay, indicating the correct assembly of the catalytic center in the cyanobacterial host. Using a similar expression system, the hydrogenase structural genes from Thiocapsa roseopersicina (hynSL) and the entire set of known accessory genes were transferred to S. elongatus. A protein of the correct size was expressed but had no activity. However, when the 11 accessory genes from AltDE were co-expressed with hynSL, the T. roseopersicina hydrogenase was found to be active by in vitro assay. This is the first report of active, heterologously-expressed [NiFe] hydrogenases in cyanobacteria. PMID- 21637847 TI - Mycoplasma genitalium: an emerging cause of sexually transmitted disease in women. AB - Mycoplasma genitalium is an emerging sexually transmitted pathogen implicated in urethritis in men and several inflammatory reproductive tract syndromes in women including cervicitis, pelvic inflammatory disease (PID), and infertility. This comprehensive review critically examines epidemiologic studies of M. genitalium infections in women with the goal of assessing the associations with reproductive tract disease and enhancing awareness of this emerging pathogen. Over 27,000 women from 48 published reports have been screened for M. genitalium urogenital infection in high- or low-risk populations worldwide with an overall prevalence of 7.3% and 2.0%, respectively. M. genitalium was present in the general population at rates between those of Chlamydia trachomatis and Neisseria gonorrhoeae. Considering more than 20 studies of lower tract inflammation, M. genitalium has been positively associated with urethritis, vaginal discharge, and microscopic signs of cervicitis and/or mucopurulent cervical discharge in seven of 14 studies. A consistent case definition of cervicitis is lacking and will be required for comprehensive understanding of these associations. Importantly, evidence for M. genitalium PID and infertility are quite convincing and indicate that a significant proportion of upper tract inflammation may be attributed to this elusive pathogen. Collectively, M. genitalium is highly prevalent in high- and low-risk populations, and should be considered an etiologic agent of select reproductive tract disease syndromes in women. PMID- 21637848 TI - The impact of subsidies on the ecological sustainability and future profits from North Sea fisheries. AB - BACKGROUND: This study examines the impact of subsidies on the profitability and ecological stability of the North Sea fisheries over the past 20 years. It shows the negative impact that subsidies can have on both the biomass of important fish species and the possible profit from fisheries. The study includes subsidies in an ecosystem model of the North Sea and examines the possible effects of eliminating fishery subsidies. METHODOLOGY/PRINCIPAL FINDINGS: Hindcast analysis between 1991 and 2003 indicates that subsidies reduced the profitability of the fishery even though gross revenue might have been high for specific fisheries sectors. Simulations seeking to maximise the total revenue between 2004 and 2010 suggest that this can be achieved by increasing the effort of Nephrops trawlers, beam trawlers, and the pelagic trawl-and-seine fleet, while reducing the effort of demersal trawlers. Simulations show that ecological stability can be realised by reducing the effort of the beam trawlers, Nephrops trawlers, pelagic- and demersal trawl-and-seine fleets. This analysis also shows that when subsidies are included, effort will always be higher for all fleets, because it effectively reduces the cost of fishing. CONCLUSIONS/SIGNIFICANCE: The study found that while removing subsidies might reduce the total catch and revenue, it increases the overall profitability of the fishery and the total biomass of commercially important species. For example, cod, haddock, herring and plaice biomass increased over the simulation when optimising for profit, and when optimising for ecological stability, the biomass for cod, plaice and sole also increased. When subsidies are eliminated, the study shows that rather than forcing those involved in the fishery into the red, fisheries become more profitable, despite a decrease in total revenue due to a loss of subsidies from the government. PMID- 21637849 TI - Pre-microRNA and mature microRNA in human mitochondria. AB - BACKGROUND: Because of the central functions of the mitochondria in providing metabolic energy and initiating apoptosis on one hand and the role that microRNA (miRNA) play in gene expression, we hypothesized that some miRNA could be present in the mitochondria for post-transcriptomic regulation by RNA interference. We intend to identify miRNA localized in the mitochondria isolated from human skeletal primary muscular cells. METHODOLOGY/PRINCIPAL FINDINGS: To investigate the potential origin of mitochondrial miRNA, we in-silico searched for microRNA candidates in the mtDNA. Twenty five human pre-miRNA and 33 miRNA aligments (E value<0.1) were found in the reference mitochondrial sequence and some of the best candidates were chosen for a co-localization test. In situ hybridization of pre-mir-302a, pre-let-7b and mir-365, using specific labelled locked nucleic acids and confocal microscopy, demonstrated that these miRNA were localized in mitochondria of human myoblasts. Total RNA was extracted from enriched mitochondria isolated by an immunomagnetic method from a culture of human myotubes. The detection of 742 human miRNA (miRBase) were monitored by RT-qPCR at three increasing mtRNA inputs. Forty six miRNA were significantly expressed (2(nd) derivative method Cp>35) for the smallest RNA input concentration and 204 miRNA for the maximum RNA input concentration. In silico analysis predicted 80 putative miRNA target sites in the mitochondrial genome (E-value<0.05). CONCLUSIONS/SIGNIFICANCE: The present study experimentally demonstrated for the first time the presence of pre-miRNA and miRNA in the human mitochondria isolated from skeletal muscular cells. A set of miRNA were significantly detected in mitochondria fraction. The origin of these pre-miRNA and miRNA should be further investigate to determine if they are imported from the cytosol and/or if they are partially processed in the mitochondria. PMID- 21637850 TI - Human macrophages infected with a high burden of ESAT-6-expressing M. tuberculosis undergo caspase-1- and cathepsin B-independent necrosis. AB - Mycobacterium tuberculosis (Mtb) infects lung macrophages, which instead of killing the pathogen can be manipulated by the bacilli, creating an environment suitable for intracellular replication and spread to adjacent cells. The role of host cell death during Mtb infection is debated because the bacilli have been shown to be both anti-apoptotic, keeping the host cell alive to avoid the antimicrobial effects of apoptosis, and pro-necrotic, killing the host macrophage to allow infection of neighboring cells. Since mycobacteria activate the NLRP3 inflammasome in macrophages, we investigated whether Mtb could induce one of the recently described inflammasome-linked cell death modes pyroptosis and pyronecrosis. These are mediated through caspase-1 and cathepsin-B, respectively. Human monocyte-derived macrophages were infected with virulent (H37Rv) Mtb at a multiplicity of infection (MOI) of 1 or 10. The higher MOI resulted in strongly enhanced release of IL-1beta, while a low MOI gave no IL-1beta response. The infected macrophages were collected and cell viability in terms of the integrity of DNA, mitochondria and the plasma membrane was determined. We found that infection with H37Rv at MOI 10, but not MOI 1, over two days led to extensive DNA fragmentation, loss of mitochondrial membrane potential, loss of plasma membrane integrity, and HMGB1 release. Although we observed plasma membrane permeabilization and IL-1beta release from infected cells, the cell death induced by Mtb was not dependent on caspase-1 or cathepsin B. It was, however, dependent on mycobacterial expression of ESAT-6. We conclude that as virulent Mtb reaches a threshold number of bacilli inside the human macrophage, ESAT-6-dependent necrosis occurs, activating caspase-1 in the process. PMID- 21637852 TI - MosSCI and gateway compatible plasmid toolkit for constitutive and inducible expression of transgenes in the C. elegans germline. AB - Here we describe a toolkit for the production of fluorescently tagged proteins in the C. elegans germline and early embryo using Mos1-mediated single copy insertion (MosSCI) transformation. We have generated promoter and 3'UTR fusions to sequences of different fluorescent proteins yielding constructs for germline expression that are compatible with MosSCI MultiSite Gateway vectors. These vectors allow tagged transgene constructs to be inserted as single copies into known sites in the C. elegans genome using MosSCI. We also show that two C. elegans heat shock promoters (Phsp-16.2 and Phsp-16.41) can be used to induce transgene expression in the germline when inserted via MosSCI transformation. This flexible set of new vectors, available to the research community in a plasmid repository, should facilitate research focused on the C. elegans germline and early embryo. PMID- 21637851 TI - p21(WAF1/CIP1) upregulation through the stress granule-associated protein CUGBP1 confers resistance to bortezomib-mediated apoptosis. AB - BACKGROUND: p21(WAF1/CIP1) is a well known cyclin-dependent kinase inhibitor induced by various stress stimuli. Depending on the stress applied, p21 upregulation can either promote apoptosis or prevent against apoptotic injury. The stress-mediated induction of p21 involves not only its transcriptional activation but also its posttranscriptional regulation, mainly through stabilization of p21 mRNA levels. We have previously reported that the proteasome inhibitor MG132 induces the stabilization of p21 mRNA, which correlates with the formation of cytoplasmic RNA stress granules. The mechanism underlying p21 mRNA stabilization, however, remains unknown. METHODOLOGY/PRINCIPAL FINDINGS: We identified the stress granules component CUGBP1 as a factor required for p21 mRNA stabilization following treatment with bortezomib ( = PS-341/Velcade). This peptide boronate inhibitor of the 26S proteasome is very efficient for the treatment of myelomas and other hematological tumors. However, solid tumors are sometimes refractory to bortezomib treatment. We found that depleting CUGBP1 in cancer cells prevents bortezomib-mediated p21 upregulation. FISH experiments combined to mRNA stability assays show that this effect is largely due to a mistargeting of p21 mRNA in stress granules leading to its degradation. Altering the expression of p21 itself, either by depleting CUGBP1 or p21, promotes bortezomib-mediated apoptosis. CONCLUSIONS/SIGNIFICANCE: We propose that one key mechanism by which apoptosis is inhibited upon treatment with chemotherapeutic drugs might involve upregulation of the p21 protein through CUGBP1. PMID- 21637853 TI - A linear model for transcription factor binding affinity prediction in protein binding microarrays. AB - Protein binding microarrays (PBM) are a high throughput technology used to characterize protein-DNA binding. The arrays measure a protein's affinity toward thousands of double-stranded DNA sequences at once, producing a comprehensive binding specificity catalog. We present a linear model for predicting the binding affinity of a protein toward DNA sequences based on PBM data. Our model represents the measured intensity of an individual probe as a sum of the binding affinity contributions of the probe's subsequences. These subsequences characterize a DNA binding motif and can be used to predict the intensity of protein binding against arbitrary DNA sequences. Our method was the best performer in the Dialogue for Reverse Engineering Assessments and Methods 5 (DREAM5) transcription factor/DNA motif recognition challenge. For the DREAM5 bonus challenge, we also developed an approach for the identification of transcription factors based on their PBM binding profiles. Our approach for TF identification achieved the best performance in the bonus challenge. PMID- 21637854 TI - Expression of miRNAs miR-133b and miR-206 in the Il17a/f locus is co-regulated with IL-17 production in alphabeta and gammadelta T cells. AB - Differentiation of T helper 17 cells (Th17) is a multistep process that involves the cytokines IL-6, TGF-beta, and IL-23 as well as IL-1beta, IL-21, and TNF alpha. Thereby, robust induction of the capacity to produce IL-17 involves epigenetic modifications of the syntenic Il17a/f locus. Using inbred mouse strains, we identified co-regulation of gene transcription at the Il17a/f locus with the nearby microRNAs miR-133b and miR-206 that are clustered approximately 45 kb upstream of Il17a/f. Expression of these microRNAs was specific for Th17 as compared to other CD4(+) T cell subsets and this was equally valid for in vitro polarized and ex vivo derived cells. From all factors analyzed, IL-23 was the most important cytokine for the in vitro induction of miR-133b and miR-206 in naive CD4(+) T cells of wild type mice. However, analysis of IL-23R deficient mice revealed that IL-23R signaling was not essential for the induction of miR 133b and miR-206. Importantly, we found a similar co-regulation in CCR6(+) and other gammadelta T cell subsets that are predisposed to production of IL-17. Taken together, we discovered a novel feature of T cell differentiation towards an IL-17-producing phenotype that is shared between alphabeta and gammadelta T cells. Notably, the specific co-regulation of miR-133b and miR-206 with the Il17a/f locus also extended to human Th17 cells. This qualifies expression of miR 133b and miR-206 in T cells as novel biomarkers for Th17-type immune reactions. PMID- 21637855 TI - Gene network landscape of the ciliate Tetrahymena thermophila. AB - BACKGROUND: Genome-wide expression data of gene microarrays can be used to infer gene networks. At a cellular level, a gene network provides a picture of the modules in which genes are densely connected, and of the hub genes, which are highly connected with other genes. A gene network is useful to identify the genes involved in the same pathway, in a protein complex or that are co-regulated. In this study, we used different methods to find gene networks in the ciliate Tetrahymena thermophila, and describe some important properties of this network, such as modules and hubs. METHODOLOGY/PRINCIPAL FINDINGS: Using 67 single channel microarrays, we constructed the Tetrahymena gene network (TGN) using three methods: the Pearson correlation coefficient (PCC), the Spearman correlation coefficient (SCC) and the context likelihood of relatedness (CLR) algorithm. The accuracy and coverage of the three networks were evaluated using four conserved protein complexes in yeast. The CLR network with a Z-score threshold 3.49 was determined to be the most robust. The TGN was partitioned, and 55 modules were found. In addition, analysis of the arbitrarily determined 1200 hubs showed that these hubs could be sorted into six groups according to their expression profiles. We also investigated human disease orthologs in Tetrahymena that are missing in yeast and provide evidence indicating that some of these are involved in the same process in Tetrahymena as in human. CONCLUSIONS/SIGNIFICANCE: This study constructed a Tetrahymena gene network, provided new insights to the properties of this biological network, and presents an important resource to study Tetrahymena genes at the pathway level. PMID- 21637856 TI - Risk factors for hospitalized seasonal influenza in rural western Kenya. AB - BACKGROUND: Risk factors for influenza hospitalization in Africa are unknown, including the role of HIV. METHODS: We conducted a case-control study of risk factors for hospitalized seasonal influenza among persons in rural western Kenya, a high HIV prevalence area, from March 2006-August 2008. Eligible cases were >=five years old, admitted to health facilities with respiratory symptoms, and had nasopharyngeal/oropharyngeal swab specimens that tested positive for influenza A or B by real-time reverse transcription-PCR. Three randomly selected age-, sex- and neighborhood-matched controls were enrolled per case. A structured questionnaire was administered and home-based HIV testing was performed. Risk factors were evaluated using conditional logistic regression. RESULTS: A total of 64 cases (38 with influenza A and 26 with influenza B) and 190 controls were enrolled. The median age was 16 years (range 5-69 years). Among cases, 24.5% were HIV-infected versus 12.5% of controls (p = 0.004). Among persons >=18 years old, 13 (59%) of 22 tested cases were HIV-positive compared with 15 (24%) of 62 tested controls (p = 0.005). In multivariable analysis, HIV-infection was associated with hospitalization due to influenza [adjusted Odds Ratio (aOR) 3.56, 95% CI 1.25-10.1]. The mean CD4 count among HIV-infected cases and controls was similar (399 vs. 387, respectively, p = 0.89). Chronic lung disease (aOR 6.83, 95% CI 1.37-34.0) was also associated with influenza hospitalization in multivariable analysis. Active pulmonary tuberculosis was associated with influenza hospitalization in bivariate, but not multivariable, analysis. CONCLUSIONS: People with HIV infection and chronic lung disease were at increased risk of hospitalized influenza in rural Kenya. HIV infection is common in many parts of sub-Saharan Africa. Influenza vaccine might prevent severe influenza in these risk groups. PMID- 21637857 TI - lpxC and yafS are the most suitable internal controls to normalize real time RT qPCR expression in the phytopathogenic bacteria Dickeya dadantii. AB - BACKGROUND: Quantitative RT-PCR is the method of choice for studying, with both sensitivity and accuracy, the expression of genes. A reliable normalization of the data, using several reference genes, is critical for an accurate quantification of gene expression. Here, we propose a set of reference genes, of the phytopathogenic bacteria Dickeya dadantii and Pectobacterium atrosepticum, which are stable in a wide range of growth conditions. RESULTS: We extracted, from a D. dadantii micro-array transcript profile dataset comprising thirty-two different growth conditions, an initial set of 49 expressed genes with very low variation in gene expression. Out of these, we retained 10 genes representing different functional categories, different levels of expression (low, medium, and high) and with no systematic variation in expression correlating with growth conditions. We measured the expression of these reference gene candidates using quantitative RT-PCR in 50 different experimental conditions, mimicking the environment encountered by the bacteria in their host and directly during the infection process in planta. The two most stable genes (ABF-0017965 (lpxC) and ABF-0020529 (yafS) were successfully used for normalization of RT-qPCR data. Finally, we demonstrated that the ortholog of lpxC and yafS in Pectobacterium atrosepticum also showed stable expression in diverse growth conditions. CONCLUSIONS: We have identified at least two genes, lpxC (ABF-0017965) and yafS (ABF-0020509), whose expressions are stable in a wide range of growth conditions and during infection. Thus, these genes are considered suitable for use as reference genes for the normalization of real-time RT-qPCR data of the two main pectinolytic phytopathogenic bacteria D. dadantii and P. atrosepticum and, probably, of other Enterobacteriaceae. Moreover, we defined general criteria to select good reference genes in bacteria. PMID- 21637858 TI - Ciliary neurotrophic factor induces genes associated with inflammation and gliosis in the retina: a gene profiling study of flow-sorted, Muller cells. AB - BACKGROUND: Ciliary neurotrophic factor (CNTF), a member of the interleukin-6 cytokine family, has been implicated in the development, differentiation and survival of retinal neurons. The mechanisms of CNTF action as well as its cellular targets in the retina are poorly understood. It has been postulated that some of the biological effects of CNTF are mediated through its action via retinal glial cells; however, molecular changes in retinal glia induced by CNTF have not been elucidated. We have, therefore, examined gene expression dynamics of purified Muller (glial) cells exposed to CNTF in vivo. METHODOLOGY/PRINCIPAL FINDINGS: Muller cells were flow-sorted from mgfap-egfp transgenic mice one or three days after intravitreal injection of CNTF. Microarray analysis using RNA from purified Muller cells showed differential expression of almost 1,000 transcripts with two- to seventeen-fold change in response to CNTF. A comparison of transcriptional profiles from Muller cells at one or three days after CNTF treatment showed an increase in the number of transcribed genes as well as a change in the expression pattern. Ingenuity Pathway Analysis showed that the differentially regulated genes belong to distinct functional types such as cytokines, growth factors, G-protein coupled receptors, transporters and ion channels. Interestingly, many genes induced by CNTF were also highly expressed in reactive Muller cells from mice with inherited or experimentally induced retinal degeneration. Further analysis of gene profiles revealed 20-30% overlap in the transcription pattern among Muller cells, astrocytes and the RPE. CONCLUSIONS/SIGNIFICANCE: Our studies provide novel molecular insights into biological functions of Muller glial cells in mediating cytokine response. We suggest that CNTF remodels the gene expression profile of Muller cells leading to induction of networks associated with transcription, cell cycle regulation and inflammatory response. CNTF also appears to function as an inducer of gliosis in the retina. PMID- 21637859 TI - An efficient strategy for broad-range detection of low abundance bacteria without DNA decontamination of PCR reagents. AB - BACKGROUND: Bacterial DNA contamination in PCR reagents has been a long standing problem that hampers the adoption of broad-range PCR in clinical and applied microbiology, particularly in detection of low abundance bacteria. Although several DNA decontamination protocols have been reported, they all suffer from compromised PCR efficiency or detection limits. To date, no satisfactory solution has been found. METHODOLOGY/PRINCIPAL FINDINGS: We herein describe a method that solves this long standing problem by employing a broad-range primer extension-PCR (PE-PCR) strategy that obviates the need for DNA decontamination. In this method, we first devise a fusion probe having a 3'-end complementary to the template bacterial sequence and a 5'-end non-bacterial tag sequence. We then hybridize the probes to template DNA, carry out primer extension and remove the excess probes using an optimized enzyme mix of Klenow DNA polymerase and exonuclease I. This strategy allows the templates to be distinguished from the PCR reagent contaminants and selectively amplified by PCR. To prove the concept, we spiked the PCR reagents with Staphylococcus aureus genomic DNA and applied PE-PCR to amplify template bacterial DNA. The spiking DNA neither interfered with template DNA amplification nor caused false positive of the reaction. Broad-range PE-PCR amplification of the 16S rRNA gene was also validated and minute quantities of template DNA (10-100 fg) were detectable without false positives. When adapting to real-time and high-resolution melting (HRM) analytical platforms, the unique melting profiles for the PE-PCR product can be used as the molecular fingerprints to further identify individual bacterial species. CONCLUSIONS/SIGNIFICANCE: Broad range PE-PCR is simple, efficient, and completely obviates the need to decontaminate PCR reagents. When coupling with real-time and HRM analyses, it offers a new avenue for bacterial species identification with a limited source of bacterial DNA, making it suitable for use in clinical and applied microbiology laboratories. PMID- 21637860 TI - Oxidized LDL receptor 1 (OLR1) as a possible link between obesity, dyslipidemia and cancer. AB - Recent studies have linked expression of lectin-like ox-LDL receptor 1 (OLR1) to tumorigenesis. We analyzed microarray data from Olr1 knockout (KO) and wild type (WT) mice for genes involved in cellular transformation and evaluated effects of OLR1 over-expression in normal mammary epithelial cells (MCF10A) and breast cancer cells (HCC1143) in terms of gene expression, migration, adhesion and transendothelial migration. Twenty-six out of 238 genes were inhibited in tissues of OLR1 KO mice; the vast majority of OLR1 sensitive genes contained NF-kappaB binding sites in their promoters. Further studies revealed broad inhibition of NF kB target genes outside of the transformation-associated gene pool, with enrichment themes of defense response, immune response, apoptosis, proliferation, and wound healing. Transcriptome of Olr1 KO mice also revealed inhibition of de novo lipogenesis, rate-limiting enzymes fatty acid synthase (Fasn), stearoyl-CoA desaturase (Scd1) and ELOVL family member 6 (Elovl6), as well as lipolytic phospholipase A2 group IVB (Pla2g4b). In studies comparing MCF10A and HCC1143, the latter displayed 60% higher OLR1 expression. Forced over-expression of OLR1 resulted in upregulation of NF-kappaB (p65) and its target pro-oncogenes involved in inhibition of apoptosis (BCL2, BCL2A1, TNFAIP3) and regulation of cell cycle (CCND2) in both cell lines. Basal expression of FASN, SCD1 and PLA2G4B, as well as lipogenesis transcription factors PPARA, SREBF2 and CREM, was higher in HCC1143 cells. Over-expression of OLR1 in HCC1143 cells also enhanced cell migration, without affecting their adherence to TNFalpha-activated endothelium or transendothelial migration. On the other hand, OLR1 neutralizing antibody inhibited both adhesion and transmigration of untreated HCC1143 cells. We conclude that OLR1 may act as an oncogene by activation of NF-kB target genes responsible for proliferation, migration and inhibition of apoptosis and de novo lipogenesis genes. PMID- 21637861 TI - Synthesis, structure and DFT study of dinuclear iron, cobalt and nickel complexes with cyclopentadienyl-metal moieties. AB - Reactions of 1,1'-bis(dipheny1phosphino)cobaltocene with Co(PMe(3))(4), Ni(PMe(3))(4), Fe(PMe(3))(4), Ni(COD)(2), FeMe(2)(PMe(3))(4) or NiMe(2)(PMe(3))(3) afford a series of novel dinuclear complexes [((Me(3)P)[lower bond 1 start]Co(eta(5)-C(5)H(4)[upper bond 1 start]PPh(2)))((Me(3)P)M[upper bond 1 end](eta(5)-C(5)H(4)P[lower bond 1 end]Ph(2)))] (M = Co(1), Ni(2) and Fe(3)) [Co(eta(5)-C(5)H(4)[upper bond 1 start]PPh(2))(2)Ni[upper bond 1 end](COD)](4), [Co(eta(5)-C(5)H(4)[upper bond 1 start]PPh(2))(2)Ni[upper bond 1 end](PMe(3))(2)] (5) and [((Me(3)P)[lower bond 1 start]Co(Me)(eta(5)-C(5)H(4)[upper bond 1 start]PPh(2)))((Me(3)P)Fe[upper bond 1 end](Me)(eta(5)-C(5)H(4)P[lower bond 1 end]Ph(2)))] (6). Reactions of 1,1'-bis(dipheny1phosphino)ferrocene with Ni(PMe(3))(4), NiMe(2)(PMe(3))(3), or Co(PMe(3))(4) gives rise to complexes [Fe(eta(5)-C(5)H(4)[upper bond 1 start]PPh(2))(2)M[upper bond 1 end](PMe(3))(2)] (M = Ni (7), Co (8)). The complexes 1-8 were spectroscopically investigated and studied by X-ray single crystal diffraction. The possible reaction mechanisms and structural characteristics are discussed. Density functional theory (DFT) calculations strongly support the deductions. PMID- 21637862 TI - Fungal DNA and pet allergen levels in Swedish day care centers and associations with building characteristics. AB - Pet allergens and mold growth related to damp are common phenomena in day care centers in Sweden but exposure measurements of these factors are lacking. The aim of this study was to investigate the relationship between building construction and indoor environment quality in Swedish day care centers and the potential for exposure to fungi (analyzed by quantitative PCR) and animal allergens (analyzed by ELISA). Measurements were performed in 21 day care centers (103 rooms) from one municipality in Sweden, which were identified as constructions at risk of dampness (85% of the buildings) and with visible damage and mold growth (54% of the buildings). Dust samples were collected using cotton swab and Petri dishes. Total fungal DNA was detected in 99% and 100%, Aspergillus/Penicillium DNA in 54% and 68%, and Stachybotrys chartarum DNA in 4% and 9% of the investigated rooms in cotton swab and Petri dish samples, respectively. The total fungal DNA levels (Geometric Mean, GM) were 4.2 * 10(6) cell equivalents per m(2) and 2.9 * 10(5) cell equivalents per m(2) per day in the swab and Petri dish samples, respectively. The concentrations (GM) of cat (Fel d1), dog (Can f1), and horse (Equ cx) allergens were 9.4, 7.2 ng m(-2) day(-1), and 5.0 unit per m(2) per day, respectively. Total fungal DNA levels were higher in risk construction buildings (p = 0.01), in rooms with linoleum flooring material (p = 0.003), and in buildings with rotating heat exchangers (p = 0.02). There were associations between total fungal DNA levels and cat (p = 0.02), dog (p < 0.001), and horse (p = 0.001) allergens. In conclusion, risk constructions, damp constructions, mould growth, fungal DNA, and animal allergens were common exposure factors in Swedish day care centers. Building constructions that represent a high risk for internal dampness should be avoided in the future, and measures to reduce allergen levels should be considered to protect pet-allergic children from asthmatic problems. PMID- 21637863 TI - Wearable electrochemical sensors for in situ analysis in marine environments. AB - The development of wearable screen-printed electrochemical sensors on underwater garments comprised of the synthetic rubber neoprene is reported. These wearable sensors are able to determine the presence of environmental pollutants and security threats in marine environments. Owing to its unique elastic and superhydrophobic morphology, neoprene is an attractive substrate for thick-film electrochemical sensors for aquatic environments and offers high-resolution printing with no apparent defects. The neoprene-based sensor was evaluated for the voltammetric detection of trace heavy metal contaminants and nitroaromatic explosives in seawater samples. We also describe the first example of enzyme (tyrosinase) immobilization on a wearable substrate towards the amperometric biosensing of phenolic contaminants in seawater. Furthermore, the integration of a miniaturized potentiostat directly on the underwater garment is demonstrated. The wearable sensor-potentiostat microsystem provides a visual indication and alert if the levels of harmful contaminants have exceeded a pre-defined threshold. The concept discussed here is well-suited for integration into dry- and wetsuits worn by divers and recreational surfers/swimmers, thereby providing them with the ability to continuously assess their surroundings for environmental contaminants and security hazards. PMID- 21637864 TI - Quantum dot-sensitized solar cells incorporating nanomaterials. AB - Quantum dot-sensitized solar cells (QDSSCs) are interesting energy devices because of their (i) impressive ability to harvest sunlight and generate multiple electron/hole pairs, (ii) ease of fabrication, and (iii) low cost. The power conversion efficiencies (eta) of most QDSSCs (typically <4%) are, however, less than those (up to 12%) of dye-sensitized solar cells, mainly because of narrow absorption ranges and charge recombination occurring at the QD-electrolyte and TiO(2)-electrolyte interfaces. To further increase the values of eta of QDSSCs, it will be necessary to develop new types of working electrodes, sensitizers, counter electrodes and electrolytes. This Feature Article describes the nanomaterials that have been used recently as electronic conductors, sensitizers and counter electrodes in QDSSCs. The nature, size, morphology and quantity of these nanomaterials all play important roles affecting the efficiencies of electron injection and light harvesting. We discuss the behavior of several important types of semiconductor nanomaterials (sensitizers, including CdS, Ag(2)S, CdSe, CdTe, CdHgTe, InAs and PbS) and nanomaterials (notably TiO(2), ZnO and carbon-based species) that have been developed to improve the electron transport efficiency of QDSSCs. We point out the preparation of new generations of nanomaterials for QDSSCs and the types of electrolytes, particularly iodide/triiodide electrolytes (I(-)/I(3)(-)), polysulfide electrolytes (S(2 )/S(x)(2-)), and cobalt redox couples ([Co(o-phen)(3)(2+)/(3+)]), that improve their lifetimes. With advances in nanotechnology, we foresee significant improvements in the efficiency (eta > 6%) and durability (>3000 h) of QDSSCs. PMID- 21637865 TI - Template-free synthesis of silica ellipsoids. AB - We have created ellipsoidal or spherical morphologies of silica particles in a template-free scheme that involves controlling surface tension forces through selected volume ratios of a water/oil micellar system. PMID- 21637866 TI - Evidence for the optical signalling of changes in bicarbonate concentration within the mitochondrial region of living cells. AB - Image and spectral intensity from bicarbonate-selective europium(III) probes localised in the mitochondria of cells is modulated reversibly by variation of external pCO(2), and is suppressed by addition of the carbonic anhydrase inhibitor, acetazolomide. PMID- 21637867 TI - Covalent diamond-gold nanojewel hybrids via photochemically generated carbenes. AB - Photochemical generation of a reactive carbene-modified gold nanoparticle (AuNP) in the presence of micro-diamond monocrystalline powder leads to a robust covalent diamond-AuNP hybrid material. PMID- 21637868 TI - A combined experimental inelastic neutron scattering, Raman and ab initio lattice dynamics study of alpha-lithium amidoborane. AB - A combination of inelastic neutron scattering (INS) spectroscopy and Raman spectroscopy with periodic density functional theory calculations is used to provide a complete assignment of the vibrational spectra of alpha-lithium amidoborane (alpha-LiNH(2)BH(3)). The Born charge density and the atomic motion up to the decomposition temperature have been modelled. These models not only explain the nature of bonding in alpha-LiNH(2)BH(3) but also provide an insight into the atomic mechanisms of its decomposition. The (INS) measurements were performed in the range of 0-4000 cm(-1) on the high-resolution time-of-flight TOSCA INS spectrometer at the ISIS Spallation Neutron Source at the Rutherford Appleton Laboratory. PMID- 21637869 TI - Influence of clustering and molecular orbital shapes on the ionization enhancement in ammonia. AB - The Coulomb explosion of clusters is known to be an efficient source for producing multiply charged ions through an enhanced ionization process. However, the factors responsible for obtaining these high charge states have not been previously explored in detail and remain poorly understood. By comparing intensity-resolved visible laser excitation experiments with semi-classical theory over a range spanning both multiphoton and tunneling ionization regimes, we reveal the mechanism in which extreme ionization proceeds. Under laser conditions that can only singly ionize individual molecules, ammonia clusters generate ions depleted of all valence electrons. The geometries of the molecular orbitals are revealed to be important in driving the ionization, and can be entirely emptied at the energy requirement for removal of the first electron in the orbital. The results are in accord with non-sequential ionization arising from electrons tunneling from three separate molecular orbitals aided through the ionization ignition mechanism. PMID- 21637870 TI - Adsorption of DNA/RNA nucleobases on hexagonal boron nitride sheet: an ab initio study. AB - Our ab initio calculations indicate that the interaction of deoxyribonucleic/ribonucleic acid (DNA/RNA) nucleobases [guanine (G), adenine (A), thymine (T), cytosine (C), and uracil (U)] with the hexagonal boron nitride (h-BN) sheet, a polar but chemically inert surface, is governed by mutual polarization. Unlike the case of graphene, all nucleobases exhibit the same stacking arrangement on the h-BN sheet due to polarization effects: the anions (N and O atoms) of nucleobases prefer to stay on top of cations (B) of the substrate as far as possible, regardless of the biological properties of nucleobases. The adsorption energies, ranging from 0.5 eV to 0.69 eV, increase in the order of U, C, T, A and G, which can be attributed to different side groups or atoms of nucleobases. The fundamental nature of DNA/RNA nucleobases and h-BN sheet remains unchanged upon adsorption, suggesting that the h-BN sheet is a promising template for DNA/RNA-related research, such as self-assembly. PMID- 21637871 TI - Weak interactions in ion-ligand complexes of C3H3(+) isomers: competition between H-bound and C-bound structures in c-C3H3(+).L and H2CCCH(+).L (L = Ne, Ar, N2, CO2, and O2). AB - Explicitly correlated coupled cluster theory at the CCSD(T)-F12x level (T. B. Adler, G. Knizia, and H.-J. Werner, J. Chem. Phys.127, 221106, 2007) has been employed to study structures and vibrations of complexes of type c-C(3)H(3)(+).L and H(2)C(3)H(+).L (L = Ne, Ar, N(2), CO(2), and O(2)). Both cations have different binding sites, allowing for the formation of weak to moderately strong hydrogen bonds as well as "C-bound" or "pi-bound" structures. In contrast to previous expectations, the energetically most favourable structures of all H(2)C(3)H(+).L complexes investigated are "C-bound", with the ligand bound to the methylenic carbon atom. The theoretical predictions enable a more detailed interpretation of infrared photodissociation (IRPD) spectra than was possible hitherto. In particular, the bands observed in the range 3238-3245 cm(-1) (D. Roth and O. Dopfer, Phys. Chem. Chem. Phys.4, 4855, 2002) are assigned to essentially free acetylenic CH stretching vibrations of the propargyl cation in "C-bound" H(2)C(3)H(+).L complexes. PMID- 21637872 TI - Initial stages of salt crystal dissolution determined with ab initio molecular dynamics. AB - The initial stages of NaCl dissolution in liquid water have been examined with state-of-the-art ab initio molecular dynamics and free energy sampling techniques. Our simulations reveal a complex multi-step process triggered by the departure of Cl ions from the lattice, with a well-defined intermediate state wherein departing ions are partially solvated but remain in contact with the crystal. The polarizability of Cl(-) is identified as the source of the anion's preferential initial dissolution, an effect which leads a forcefield based description of NaCl dissolution to fail to identify a preference for Cl over Na dissolution. PMID- 21637873 TI - The use of enzymes for construction of DNA-based objects and assemblies. AB - DNA has found wide applications in DNA-based nanotechnology due to its simplicity and predictability of its secondary structure. Selecting DNA for the nanoconstruction of objects and assemblies bears the inherent potential for manipulations and control by DNA modifying enzymes. In this tutorial review, we present an overview of the enzyme-catalysed construction of DNA-based objects and assemblies. It is illustrated how a diversity of enzyme-based biochemical reactions are transferred in nanotechnological applications. PMID- 21637875 TI - Single crystal EPR study at 95 GHz of a large Fe based molecular nanomagnet: toward the structuring of magnetic nanoparticle properties. AB - A W-band single-crystal EPR study has been performed on a molecular cluster comprising 19 iron(III) ions bridged by oxo- hydroxide ions, Fe(19), in order to investigate magnetic nanosystems with a behavior in between the one of Magnetic NanoParticles (MNP) and that of Single Molecule Magnets (SMM). The Fe(19) has a disk-like shape: a planar Fe(7) core with a brucite (Mg(OH)(2)) structure enclosed in a "shell" of 12 Fe(III) ions. EPR and magnetic measurements revealed an S = 35/2 ground state with an S = 33/2 excited state lying ~ 8 K above. The presence of other low-lying excited states was also envisaged. Rhombic Zero Field Splitting (ZFS) tensors were determined, the easy axes lying in the Fe(19) plane for both the multiplets. At particular temperatures and orientations, a partially resolved fine structure could be observed which could not be distinguished in powder spectra, due to orientation disorder. The similarities of the EPR behavior of Fe(19) and MNP, together with the accuracy of single crystal analysis, helped to shed light on spectral features observed in MNP spectra, that is a sharp line at g = 2 and a low intensity transition at g = 4. Moreover, a theoretical analysis has been used to estimate the contribution to the total magnetic anisotropy of core and surface; this latter is crucial in determining the easy axis-type anisotropy, alike that of MNP surface. PMID- 21637874 TI - Homoleptic 1-D iron selenolate complexes-synthesis, structure, magnetic and thermal behaviour of (1)(infinity)[Fe(SeR)2] (R=Ph, Mes). AB - The first examples of polymeric homoleptic iron chalcogenolato complexes (1)(infinity)[Fe(SePh)(2)] and (1)(infinity)[Fe(SeMes)(2)] (Ph = phenyl = C(6)H(5), Mes = mesityl = C(6)H(2)-2,4,6-(CH(3))(3)) have been both prepared by reaction of [Fe(N(SiMe(3))(2))(2)] with two equivalents of HSeR (R = Ph, Mes) while (1)(infinity)[Fe(SePh)(2)] was found to be also easily accessible through reactions of either FeCl(2), Fe(OOCCH(3))(2) or FeCl(3) with PhSeSiMe(3) in THF. In the crystal, the two compounds form one-dimensional chains with bridging selenolate ligands comprising distinctly different Fe-Se-Fe bridging angles, namely 71.15-72.57 degrees in (1)(infinity)[Fe(SePh)(2)] and 91.80 degrees in (1)(infinity)[Fe(SeMes)(2)]. Magnetic measurements supported by DFT calculations reveal that this geometrical change has a pronounced influence on the antiferromagnetic exchange interactions of the unpaired electrons along the chains in the two different compounds with a calculated magnetic exchange coupling constant of J = -137 cm(-1) in (1)(infinity)[Fe(SePh)(2)] and J = -20 cm(-1) in (1)(infinity)[Fe(SeMes)(2)]. In addition we were able to show that the ring molecule [Fe(SePh)(2)](12) which is a structural isomer of (1)(infinity)[Fe(SePh)(2)] behaves magnetically similar to the latter one. Investigations by powder XRD reveal that the ring molecule is only a metastable intermediate which converts in THF completely to form (1)(infinity)[Fe(SePh)(2)]. Thermal gravimetric analysis of (1)(infinity)[Fe(SePh)(2)] under vacuum conditions shows that the compound is thermally labile and already starts to decompose above 30 degrees C in a two step process under cleavage of SePh(2) to finally form at 250 degrees C tetragonal PbO-type FeSe. The reaction of (1)(infinity)[Fe(SePh)(2)] with the Lewis base 1,10-phenanthroline yielded, depending on the conditions, the octahedral monomeric complexes [Fe(SePh)(2)(1,10 phen)(2)] and [Fe(1,10-phen)(3)][Fe(SePh)(4)]. PMID- 21637876 TI - Multifunctional nanocomposites constructed from Fe3O4-Au nanoparticle cores and a porous silica shell in the solution phase. AB - This work is directed towards the synthesis of multifunctional nanoparticles composed of Fe(3)O(4)-Au nanocomposite cores and a porous silica shell (Fe(3)O(4) Au/pSiO(2)), aimed at ensuring the stability, magnetic, and optical properties of magnetic-gold nanocomposite simultaneously. The prepared Fe(3)O(4)-Au/pSiO(2) core/shell nanoparticles are characterized by means of TEM, N(2) adsorption desorption isotherms, FTIR, XRD, UV-vis, and VSM. Meanwhile, as an example of the applications, catalytic activity of the porous silica shell-encapsulated Fe(3)O(4)-Au nanoparticles is investigated by choosing a model reaction, reduction of o-nitroaniline to benzenediamine by NaBH(4). Due to the existence of porous silica shells, the reaction with Fe(3)O(4)-Au/pSiO(2) core/shell nanoparticles as a catalyst follows second-order kinetics with the rate constant (k) of about 0.0165 l mol(-1) s(-1), remarkably different from the first-order kinetics with the k of about 0.002 s(-1) for the reduction reaction with the core Fe(3)O(4)-Au nanoparticles as a catalyst. PMID- 21637877 TI - Tin/polypyrrole composite anode using sodium carboxymethyl cellulose binder for lithium-ion batteries. AB - A tin nanoparticle/polypyrrole (nano-Sn/PPy) composite was prepared by chemically reducing and coating Sn nanoparticles onto the PPy surface. The composite shows a much higher surface area than the pure nano-Sn reference sample, due to the porous higher surface area of PPy and the much smaller size of Sn in the nano Sn/PPy composite than in the pure tin nanoparticle sample. Poly(vinylidene fluoride) (PVDF) and sodium carboxymethyl cellulose (CMC) were also used as binders, and the electrochemical performance was investigated. The electrochemical results show that both the capacity retention and the rate capability are in the same order of nano-Sn/PPy-CMC > nano-Sn/PPy-PVDF > nano-Sn CMC > nano-Sn-PVDF. Scanning electronic microscopy (SEM) and electrochemical impedance spectroscopy (EIS) results show that CMC can prevent the formation of cracks in electrodes caused by the big volume changes during the charge-discharge process, and the PPy in the composite can provide a conducting matrix and alleviate the agglomeration of Sn nanoparticles. The present results indicate that the nano-Sn/PPy composite could be suitable for the next generation of anode materials with relatively good capacity retention and rate capability. PMID- 21637878 TI - Mono N,C,N-pincer complexes of titanium, vanadium and niobium. Synthesis, structure and catalytic activity in olefin polymerisation. AB - Transmetallation of 4,4'-bis{(2,6-bis[(dimethylamino)methyl]phenylgold)diphenyl phosphino}biphenyl (3) with MCl(4) (M = Ti, NbCl, V) in benzene gave the corresponding transition metal pincer complexes (4) and insoluble 4,4'-bis[P (chloro gold(I))diphenylphosphino]biphenyl (2), which can be quantitatively recovered and recycled. Interestingly, 3 did not react with TiCl(3). However, reaction of 2,6-bis[(dimethylamino)methyl]phenyllithium (1) with TiCl(3) resulted in formation of the novel diaryltitanium(IV) compound 5 (16% yield), comprising one N,C,N-mer bound NCN-pincer ligand and a second NCN-pincer ligand that is rearranged from a 1,2,6-isomer to a 1,2,4 one. The latter NCN-ligand is dianionic and is bidentate bonded; one of the CH(2)NMe(2) substituents (para to C'(ipso)) is non-coordinated, while the second CH(2)NMe(2) group, after C-H activation of one of the Me groups, is eta(2)-C,N-bonded to the titanium centre trans to C(ipso) of the mer-NCN ligand. The new NCN-pincer metal complexes 2,6 bis[(dimethylamino)methyl]phenylTiCl(3) (4a) and 2,6-bis[(dimethylamino)methyl] phenylVCl(2) (4d) gave, after immobilization on MgCl(2)-based supports, very high activity in ethene polymerisation. PMID- 21637879 TI - The first binuclear sandwich-like complexes based on the aromatic tetraatomic species. AB - The first binuclear sandwich-like complexes based on the aromatic tetraatomic species with formula M(2)(eta(4)-E(4))(2) (M = Al, Ga; E = N, P, As) have been studied by density functional theory (DFT). The stable conformer for each M(2)(eta(4)-E(4))(2) is the staggered one with D(4d) symmetry except for Ga(2)(eta(2)-N(4))(2) with C(2v) symmetry. Natural bonding orbital (NBO) analysis indicates that the metal-metal bonds of Al(2)(eta(4)-E(4))(2) (E = N, P, As) and Ga(2)(eta(4)-E(4))(2) (E = P, As) are all sigma single bonds, which are derived mostly from the s and p(z) orbitals of the metal atoms by molecular orbital (MO) analysis. For M(2)(eta(4)-E(4))(2) (M = Al, Ga; E = P, As), the metal-ligand interactions are covalent, while for Al(2)(eta(4)-N(4))(2) the interactions between the Al atoms and the N(4)(2-) ligands are ionic. According to the calculated dissociation energies for breaking metal-metal bonds, the Al-Al and Ga Ga bonds are very strong indicating that these stable sandwich-like compounds Al(2)(eta(4)-E(4))(2) (E = N, P, As) and Ga(2)(eta(4)-E(4))(2) (E = P, As) may be synthesized in future experiments. The nitrogen-rich compounds Al(2)(eta(4) N(4))(2) and Ga(2)(eta(2)-N(4))(2) may be used as potential candidates of high energy density materials (HEDMs). Nucleus-independent chemical shifts (NICS) values reveal that the E(4)(2-) rings in the Al(2)(eta(4)-E(4))(2) (E = N, P, As) and Ga(2)(eta(4)-E(4))(2) (E = P, As) species possess conflicting aromaticity (sigma antiaromaticity and pi aromaticity) and with the same ligands, the E(4)(2 ) ligands in Ga(2)(eta(4)-E(4))(2) have more aromaticity than those in Al(2)(eta(4)-E(4))(2). PMID- 21637880 TI - Visualising the equilibrium distribution and mobility of organic contaminants in soil using the chemical partitioning space. AB - Assessing the behaviour of organic chemicals in soil is a complex task as it is governed by the physical chemical properties of the chemicals, the characteristics of the soil as well as the ambient conditions of the environment. The chemical partitioning space, defined by the air-water partition coefficient (K(AW)) and the soil organic carbon-water partition coefficient (K(OC)), was employed to visualize the equilibrium distribution of organic contaminants between the air-filled pores, the pore water and the solid phases of the bulk soil and the relative importance of the three transport processes removing contaminants from soil (evaporation, leaching and particle erosion). The partitioning properties of twenty neutral organic chemicals (i.e. herbicides, pharmaceuticals, polychlorinated biphenyls and volatile chemicals) were estimated using poly-parameter linear free energy relationships and superimposed onto these maps. This allows instantaneous estimation of the equilibrium phase distribution and mobility of neutral organic chemicals in soil. Although there is a link between the major phase and the dominant transport process, such that chemicals found in air-filled pore space are subject to evaporation, those in water-filled pore space undergo leaching and those in the sorbed phase are associated with particle erosion, the partitioning coefficient thresholds for distribution and mobility can often deviate by many orders of magnitude. In particular, even a small fraction of chemical in pore water or pore air allows for evaporation and leaching to dominate over solid phase transport. Multiple maps that represent soils that differ in the amount and type of soil organic matter, water saturation, temperature, depth of surface soil horizon, and mineral matters were evaluated. PMID- 21637881 TI - Mesoporous silica as a membrane for ultra-thin implantable direct glucose fuel cells. AB - The design, fabrication and characterization of an inorganic catalyst based direct glucose fuel cell using mesoporous silica coating as a functional membrane is reported. The desired use of mesoporous silica based direct glucose fuel cell is for a blood vessel implantable device. Blood vessel implantable direct glucose fuel cells have access to higher continuous glucose concentrations. However, reduction in the implant thickness is required for application in the venous system as part of a stent. We report development of an implantable device with a platinum thin-film (thickness: 25 nm) deposited on silicon substrate (500 MUm) to serve as the anode, and graphene pressed on a stainless steel mesh (175 MUm) to serve as the cathode. Control experiments involved the use of a surfactant-coated polypropylene membrane (50 MUm) with activated carbon (198 MUm) electrodes. We demonstrate that a mesoporous silica thin film (270 nm) is capable of replacing the conventional polymer based membranes with an improvement in the power generated over conventional direct glucose fuel cells. PMID- 21637882 TI - beta-Galactosidase-instructed formation of molecular nanofibers and a hydrogel. AB - Here we report the first example of using beta-galactosidase to trigger the formation of cell compatible, supramolecular nanofibers, which ultimately may lead to a new approach for the development of soft nanotechnology. PMID- 21637883 TI - Fast and efficient microwave-assisted synthesis of functionalized peptoids via Ugi reactions. AB - A wide range of N-alkylglycines (peptoids) can be efficiently prepared via Ugi reactions using microwave irradiations. The results confirm the versatility and efficiency of the methodology for the preparation of functionalized peptoids. The products can be used in consecutive Ugi reactions to yield cyclic peptoids of potential biological interest. PMID- 21637884 TI - Asymmetric photocycloaddition of naphthamide with a diene using the provisional molecular chirality in a chiral crystal. AB - N-(2-Methoxy-1-naphthoyl)piperidine afforded chiral crystals by spontaneous crystallization, and the molecular chirality of the crystals was retained after dissolving them in a cooled solvent. An asymmetric photocycloaddition reaction with a diene was performed using the provisional chiral molecular conformation derived from these chiral crystals. PMID- 21637885 TI - Highly efficient photocatalytic dehalogenation of organic halides on TiO2 loaded with bimetallic Pd-Pt alloy nanoparticles. AB - UV irradiation of TiO(2) loaded with bimetallic Pd-Pt alloy particles promotes highly efficient dehalogenation of organic halides with alcohol as a hydrogen source. PMID- 21637886 TI - Electrochemiluminescence resonance energy transfer between an emitter electrochemically generated by luminol as the donor and luminescent quantum dots as the acceptor and its biological application. AB - Novel electrochemiluminescence resonance energy transfer (ECRET) between an emitter electrochemically generated by luminol as the donor and luminescent quantum dots as the acceptor is investigated. The ECRET technique can be used to study the interactions and conformational changes of proteins. PMID- 21637887 TI - Catalytic asymmetric hydroboration of beta,gamma-unsaturated Weinreb amides: striking influence of the borane. AB - Subtle differences in the structure of the borane strongly influence the catalytic efficiency and level of enantioselectivity in the catalytic asymmetric hydroboration of beta,gamma-unsaturated Weinreb amides. PMID- 21637888 TI - Access to unusual polycyclic spiro-enones from 2,2'-bis(allyloxy)-1,1' binaphthyls using Grubbs' catalysts: an unprecedented one-pot RCM/Claisen sequence. AB - Treatment of 2,2'-bis(allyloxy)-1,1'-binaphthyls with the first-generation Grubbs' carbene under MW-irradiation results in the formation of new polycyclic spiro-enones through an unprecedented RCM/Claisen sequence. PMID- 21637889 TI - Multifunctional water-soluble molecular capsules based on p-phosphonic acid calix[5]arene. AB - p-Phosphonic acid calix[5]arene forms molecular capsules in water based on two of the molecules, which can be loaded with carboplatin using intense shearing, and attached to single wall carbon nano-tubes. Spin coating of the capsules onto a substrate affords 2 nm fibres of stacked calixarenes, with the self-assembly understood using molecular modelling. PMID- 21637890 TI - Efficient recycling of a chiral palladium catalytic system for asymmetric allylic substitutions in ionic liquid. AB - Chiral carbohydrate-based diphosphites were used for Pd-catalysed asymmetric allylic substitution (alkylation, amination, phosphination) in neat ionic liquids (ILs). Pyrrolidinium-based IL led to the best activities, allowing an efficient catalyst immobilization. In the allylic amination (TOF > 3100 h(-1)), the catalyst could be recycled nine times preserving both activity and enantioselectivity. PMID- 21637891 TI - Au-catalyzed synthesis of 2-alkylindoles from N-arylhydroxylamines and terminal alkynes. AB - The first gold-catalyzed addition of N-arylhydroxylamines to aliphatic terminal alkynes is developed to access O-alkenyl-N-arylhydroxylamines, which undergo facile in situ sequential 3,3-rearrangements and cyclodehydrations to afford 2 alkylindoles with regiospecificity and under exceptionally mild reaction conditions. PMID- 21637892 TI - A novel catalyst-free synthesis of vertically aligned silicon nanowire-carbon nanotube heterojunction arrays for high performance electron field emitters. AB - A novel, catalyst-free strategy for the direct synthesis of vertically aligned silicon nanowire-carbon nanotube (SiNW-CNT) heterojunction arrays is presented. Such a heterojunction with the junction area in the nanoscale displays enhanced field emission characteristics at low turn-on field, with a nearly three times increase in the field enhancement factor. PMID- 21637893 TI - Flow-alignment of bicellar lipid mixtures: orientations of probe molecules and membrane-associated biomacromolecules in lipid membranes studied with polarized light. AB - Bicelles are excellent membrane-mimicking hosts for a dynamic and structural study of solutes with NMR, but the magnetic fields required for their alignment are hard to apply to optical conditions. Here we demonstrate that bicellar mixtures can be aligned by shear forces in a Couette flow cell, to provide orientation of membrane-bound retinoic acid, pyrene and cytochrome c (cyt c) protein, conveniently studied with linear dichroism spectroscopy. PMID- 21637894 TI - Perspectives on the structural and biosynthetic interrelationships between oxygenated furanocembranoids and their polycyclic congeners found in corals. AB - Macrocyclic and polycyclic 'cembranoid' diterpenes are one of the most widespread groups of natural products that are found in the marine milieu. The macrocyclic cembranoids are linked to each other by a network of oxygenation processes, which often climax with the formation of furano- and furanobutenolide-based macrocyclic cembranoids commonly found in gorgonian and soft corals. These macrocycles are then prone to oxidative rearrangements, photochemical ring contraction, and transannular cyclisations amongst others, leading to a plethora of novel and architecturally attractive marine metabolites. Although there is a dearth of knowledge about the enzymes that trigger some of the steps in their biosynthesis, speculations are rife. In this personal perspective we have examined many of the structural relationships within oxycembranoids and their relatives isolated from corals. This has allowed us to speculate on the likely biosynthetic interrelationships between structurally similar metabolites, and then propose some likely key carbon-to-carbon bond forming reactions that are followed in vivo in linking macrocyclic cembranoids to their polycyclic congeners. Biomimetic synthesis studies, which vindicate some of the biosynthetic speculations, are interweaved in the discussion. PMID- 21637895 TI - Why are dermatologists still talking about acne? Because so many people have it... and we are always seeking better ways to manage it. PMID- 21637896 TI - Why are Dermatologists Still Talking About Acne? Because So Many People Have It... and We Are Always Seeking Better Ways to Manage It. PMID- 21637898 TI - Pathogenesis of acne vulgaris: what's new, what's interesting and what may be clinically relevant. AB - Acne vulgaris is the most common skin disorder seen in dermatology and primary care offices today with significant associated morbidity. The pathogenesis of acne is complex and multifactorial, and there continues to be an influx of new information to increase our understanding of this chronic disease. Recent advances in acne pathogenesis will be discussed, including theories regarding the sequence of events in acne formation, the functions of P. acnes, TLR involvement and role of the sebaceous gland and factors influencing sebum production. PMID- 21637899 TI - Efficacy and safety of azelaic acid (AzA) gel 15% in the treatment of post inflammatory hyperpigmentation and acne: a 16-week, baseline-controlled study. AB - Although there are few differences in the incidence and pathophysiology of acne across various races and ethnicities, there is some evidence that black patients may have larger sebaceous glands and increased sebum production. Of greater clinical relevance, patients with darker skin types are at increased risk for the development of post-inflammatory hyperpigmentation (PIH), which some find as or more troubling than acne itself. This common and bothersome sequelum of acne can be difficult to manage in this population. Topical azelaic acid gel is recognized to have anti-tyrosinase activity, suggesting it may be a suitable treatment option for mild-to-moderate acne with associated moderate-to-severe PIH. This pilot study demonstrates the efficacy of topical AzA gel 15% when applied twice daily for the reduction of both acne and PIH. J PMID- 21637900 TI - Evaluation of self-treatment of mild-to-moderate facial acne with a blue light treatment system. AB - INTRODUCTION: This study evaluated the efficacy and tolerability of treating mild to-moderate facial acne using a new, hand-held, light-emitting diode blue light device in conjunction with a foam cleanser containing 5% glycolic acid and 2% salicylic acid plus a skin rebuilding serum containing 1.25% salicylic acid, 0.5% niacinamide, 0.08% liposomal-based azelaic acid and superoxide dismutase. METHODS: Volunteers with mild-to-moderate facial inflammatory acne used the blue light device twice daily for eight weeks, plus the cleanser before treatments and the serum after each evening treatment. RESULTS: Among 33 subjects aged 25-45 years old, 28 completed. In a 3 cm x 5 cm target area receiving a daily dose of ~29 J/cm2, treatment was associated with significant reductions from baseline in the inflammatory lesion count from week 1 onward (P<= .01) and in the non inflammatory lesion count from week 4 onward (P<= .05). The number of flares was significantly reduced from baseline from week 2 onward (P<= .05), and flare severity and flare redness were significantly reduced from baseline from week 4 onward (P<= .01 and P<= .05, respectively). At week 8, more than 90 percent of subjects reported improvements in their skin's overall appearance, clarity, radiance, tone, texture and smoothness. In addition, 82 percent were satisfied, very satisfied, or extremely satisfied with the blue light treatment system and 86 percent agreed the treatment system was much gentler than traditional acne treatments. CONCLUSION: The blue light treatment system offers effective, rapid, convenient and well tolerated treatment of inflammatory and non-inflammatory acne lesions. The majority of subjects consider it much gentler than traditional acne treatments and it facilitates effective treatment without the need for antibiotic exposure. The blue light treatment system and blue light therapy alone are attractive treatment options for acne vulgaris, both as alternatives to traditional acne treatments and as adjunctive treatments to complement existing therapies. PMID- 21637901 TI - How patients experience and manage dryness and irritation from acne treatment. AB - BACKGROUND: Products that may cause irritation are widely used to treat acne. Irritation has the potential to reduce treatment adherence. How patients manage irritation and dryness is not well characterized. OBJECTIVES: To study self reported irritation, its impact and coping mechanisms in patients who had been treated for acne with a clindamycin-5% benzoyl peroxide (BPO) product. METHODS: An Internet-based survey of 200 subjects, aged 15-40 years who had used a clindamycin-5% BPO fixed combination product in the last six months on at least 50 percent of their face, at least five days per week. RESULTS: The majority of subjects (57%) had moderate acne, 28 percent had severe acne. Bothersome side effects of the clindamycin-5% BPO combination included dry skin (55%), flaky/peeling skin (45%), irritated skin (44%), itchy skin (39%) and redness (37%). As a result, subjects used the product only as a spot treatment (33%), only when breakouts seemed worse (28%), or less often than recommended (32%); stopped using from time to time (32%); switched to a different prescription medication and/or an over-the-counter acne product (28%); or stopped using altogether (10%). 41 percent of subjects reported using moisturizers to counteract dryness and redness. LIMITATIONS: We queried patients concerning use of combination clindamycin/BPO products and not other products. DISCUSSION: Irritation to clindamycin-5% BPO is a common problem that reduces patients' use of the medication. Strategies to improve treatment include communication with patients on possible side effects, providing written instruction on how to manage irritation and dryness and consideration of alternative topical treatments and treatment regimens. PMID- 21637902 TI - Nephrogenic systemic fibrosis: a case report and review of the literature. AB - The authors report a case of a patient who presented with thick, indurated, hyperpigmented plaques of the bilateral upper and lower extremities, ultimately discovered to be attributed to nephrogenic systemic fibrosis. The case was written to highlight the clinical manifestations of nephrogenic systemic fibrosis, as well as to make dermatologists aware of this disease, their role in diagnosis and management, and review treatment options. PMID- 21637903 TI - Topical rosacea therapy: the importance of vehicles for efficacy, tolerability and compliance. AB - Many topical medications are available for the treatment of papulopustular rosacea. While treatments contain metronidazole, azelaic acid, or sodium sulfacetamide-sulfur as the active ingredient, the composition of the vehicle formulations varies widely. These vehicles come in gels, creams, lotions and foams; some ingredients are common to many vehicles, while some vehicles contain unique ingredients designed to optimize skin penetration and delivery of the active drug to its target. Vehicles can also influence tolerability, which is always a concern in patients with heightened skin sensitivity, and compliance, which is typically lower for topical treatments than oral treatments. Ideally, the vehicle of any rosacea treatment should enhance drug delivery, be nonirritating and be easy to use. Ingredients that help repair barrier function are also desirable. This review will focus on the key components of the vehicles from the most commonly used topical therapies for papulopustular rosacea and how vehicle formulations influence the delivery of active ingredient, skin barrier repair, tolerability and compliance. PMID- 21637904 TI - Evidence-based review: fixed-combination therapy and topical retinoids in the treatment of acne. AB - Topical fixed-combination products and topical retinoid monotherapy are established first-line treatments for mild-to-moderate acne vulgaris, yet adequate comparative data are lacking. The following evidence-based review addresses the question: "In patients with mild-to-moderate acne, are topical fixed-combination products or topical retinoids a more efficacious choice in reducing noninflammatory, inflammatory and total lesions after 12 weeks of treatment?" To identify relevant studies, a PubMed search was performed using "acne" and search terms for adapalene, tretinoin, tazarotene, benzoyl peroxide, clindamycin, or erythromycin. Forty-two studies from January 1991 to November 2009 were included. The studies were evaluated using the Strength of Recommendation Taxonomy, and all but seven received the highest level of evidence grade. To evaluate efficacy, a side-by-side comparison was made using reduction in acne lesion counts at week 12 for study groups treated with fixed-combination therapy or retinoid monotherapy. Twenty-nine studies containing relevant efficacy data for fixed-combination therapy and retinoid monotherapy are summarized here. Nine studies compared fixed-combination therapy with retinoid monotherapy; in eight of these studies, fixed-combination therapy was significantly more efficacious in reducing acne lesion counts. This evidence-based review analyzes clinical evidence to date for these therapies to provide guidance in determining appropriate treatment for patients with mild-to-moderate acne. PMID- 21637905 TI - Safety and efficacy of micronized tretinoin gel (0.05%) in treating adolescent acne. AB - Tretinoin is widely used in the treatment of acne. Despite significant advances in formulation development, irritation and dryness can be particularly bothersome, especially during the first 3-4 weeks, impacting adherence. Dose titration and adjunct use of moisturizers have been commonly employed. Co prescribing with benzoyl peroxide (BPO) or a BPO/antibiotic combination is also common practice. The tretinoin molecule is unstable and can be degraded by BPO, further complicating treatment regimens. Lately, formulation technology has focused on providing more efficient penetration of the tretinoin into the skin layers so that lower concentrations of tretinoin might afford better tolerability, but maintain good efficacy; incorporating moisturizing excipients to minimize irritation; and providing greater stability to the tretinoin molecule. This approach would be particularly relevant in a pediatric acne population where efficacy/tolerability balance is important and treatment regimens must take into account lifestyles, but little data exist on the use of tretinoin in this patient population. A micronized formulation of tretinoin (0.05%) gel has been developed that provides a more efficient delivery of tretinoin, because of its optimal particle size, no degradation by BPO and better cutaneous tolerability than tretinoin microsphere (0.1%) gel without compromising efficacy in a pediatric population. PMID- 21637906 TI - Ten tips for treating acne vulgaris in Fitzpatrick skin types IV-VI. AB - During the 21st century, people with skin of color will account for a large portion of the US population. According to race projections from the US Census Bureau, persons of African American, Hispanic, Asian and Native American descent will make up nearly half of the total US population by the year 2050. Because of our changing demographics, it is important that all dermatologists become comfortable in treating skin of color. This article provides 10 practical pearls for management of acne vulgaris in patients with Fitzpatrick skin types IV-VI. PMID- 21637907 TI - Taurine chloramine inhibits NO and TNF-alpha production in zymosan plus interferon-gamma activated RAW 264.7 cells. AB - Taurine is present abundantly in various tissues, especially in leukocytes embattled to foreign invaders such as microorganisms or oxidants. Taurine chloramine (Tau-Cl) is produced from taurine at the site of inflammation via the myeloperoxidase-halide pathway in leukocytes induced by oxidants and/or infectious materials. Previously, our data demonstrated that Tau-Cl inhibited nitric oxide (NO) production and TNF-alpha secretion induced by lipopolysaccharide (LPS), a Toll-like receptor 4 (TLR-4) ligand or lipoarabinomannan (LAM), a TLR-2 ligand plus interferon-gamma (IFN-gamma) in peritoneal macrophages or RAW 264.7 cells. Zymosan, a beta-glucan of yeast cell wall, is a ligand for TLR-2 and dectin-1 and stimulates macrophages to produce proinflammatory mediators such as NO and TNF-alpha. Based on our previous data, we examined the effect of zymosan and IFN-gamma induced production of NO and TNF alpha in the absence or presence of Tau-Cl or taurine using RAW 264.7 cells. Production of NO and secretion of TNF-alpha is increased when zymosan is combined with IFN-gamma. Tau-Cl inhibited production of NO and secretion of TNF-alpha in zymosan plus IFN-gamma activated RAW 264.7 cells in a dose-dependent manner (99% vs. 48% using 0.8mM Tau-Cl). Taurine was without effect. Nitric oxide synthase protein (iNOS), induced by zymosan plus IFN-gamma, was inhibited by Tau-Cl (0.8mM) as measured using western blot analysis. NOS mRNA was inhibited by Tau-Cl at four, eight and 16 hours post activation, but not at 24 hours. TNF-alpha mRNA was inhibited at four hours and eight hours, but not at 16 and 24 hours. These data suggest that expression of both iNOS and TNF-alpha mRNAs are inhibited by treatment with Tau-Cl within four and eight hours, but not at later time points. Transient suppression of activation of RAW 264.7 cells induced by zymosan may play a critical physiological role for taurine in protecting against tissue injury from initial overt inflammation. This study indicates that tropical treatment of taurine may ameliorate inflammatory dermatoses caused by an environmental yeast or abnormal immune function. PMID- 21637908 TI - Bilateral comparison study of pimecrolimus cream 1% and a ceramide-hyaluronic acid emollient foam in the treatment of patients with atopic dermatitis. AB - Topical corticosteroids have been the mainstay of treatment for atopic dermatitis (AD) over the last decade, especially in the setting of acute flares. However, heavy and prolonged use of topical corticosteroid is undesirable as it is associated with side effects such as, skin atrophy, telangiectasia, striae, steroid-induced dermatoses, rosacea, acne exacerbation, and in some severe and rare cases, systemic effects such as hypothalamic-pituitary-adrenal axis suppression, growth retardation and ocular problems. Non-steroidal ant inflammatory agents specific for the treatment of AD (topical calcineurin inhibitors, or TCIs) are now available and they are a viable alternative to topical corticosteroids in treating dermatitis of the face, neck, eyelids, and intertriginous areas where there is a greater risk of the steroid-induced side effects. More recently, medical device emollients have entered the marketplace. These medical devices provide, but are not limited to, anti-oxidant, anti protease, anti-inflammatory activity, and aid in restoring the natural balance of lipids, which is one of the causes of the epidermal abnormalities seen with AD. The present study evaluated the short-term effectiveness and appeal of a non steroidal medicated device foam as compared to pimecrolimus cream 1% in the treatment of AD within a wide age group of subjects with active disease at baseline. In this study, both pimecrolimus and the medical device foam exhibited efficacy in mild-to-moderate AD. Primary efficacy was measured by IGA. After four weeks of treatment with the medical device foam, 82% of target lesions were scored "clear" (0) or "almost clear" (1) compared to 71% of target lesions under the pimecrolimus arm. This study confirmed that pimecrolimus cream 1% and the medical device foam work well in the treatment of AD in both adults and children with no associated adverse effects. PMID- 21637909 TI - A guide to temporal fossa augmentation with small gel particle hyaluronic acid dermal filler. AB - Loss of volume in the temple can result in a gaunt, wasted appearance. Dermal filler augmentation of the temples can counteract volume loss and achieve a more balanced and youthful appearance. Although the temporal fossa is a critical area for volume restoration of the aging face, published information is limited. The authors retrospectively describe the treatment of 20 female patients who sought facial rejuvenation and received small gel particle hyaluronic acid (SPG-HA) injections for temporal fossa augmentation. The authors discuss a rationale for their choice of dermal filler and provide a detailed, illustrated injection technique guide for restoring volume in the temporal fossa region with SPG-HA. There is a need for prospective, controlled studies investigating safety, efficacy and persistency of hyaluronic acid fillers in this area of the face. PMID- 21637910 TI - A biopsy diagnosis? Clinical clues and patterns to help distinguish cutaneous metastases: part I of II. PMID- 21637911 TI - Altered oxido-reductive state in the diabetic heart: loss of cardioprotection due to protein disulfide isomerase. AB - Diabetes is associated with an increased risk of heart failure, in part explained by endoplasmic reticulum stress and apoptosis. Protein disulfide isomerase (PDI) prevents stressed cardiomyocytes apoptosis. We hypothesized that diabetes impairs PDI function by an alteration in its oxido-reductive state. Myocardial biopsies harvested from the anterolateral left ventricular wall from diabetic (n = 7) and nondiabetic (n = 8) patients were used to assess PDI expression and cardiomyocyte death. A mouse model of diabetes (streptozotocin injection, 130 mg/mL) was used to study PDI expression and its redox state after ischemia/reperfusion injury induced by 30-min occlusion of the left anterior coronary artery followed by reperfusion. Transthoracic echocardiography was performed to assess cardiac remodeling after 1 wk. Western blot analysis was used to analyze PDI expression, and methoxy-polyethyleneglycol-maleimide was used to assess its redox state. Dehydroascorbate (DHA) administration was used to restore the PDI redox state. Diabetic patients had a greater number of transferase-mediated dUTP nick-end labeling (TUNEL)-positive cells than nondiabetic patients despite a greater myocardial PDI expression suggesting altered PDI function. Diabetic mice had a worse postinfarction remodeling associated with an altered PDI redox state. DHA treatment restored functional PDI redox state and ameliorated post-myocardial infarction remodeling. An increase in PDI levels with a paradoxical decrease of its active form occurs in the diabetic heart after ischemia and may explain the lack of protective effects of PDI in diabetes. Restoration of PDI redox state prevents adverse remodeling. The potential significance of these findings deserves to be validated in a clinical setting. PMID- 21637912 TI - Differential microRNA expression and identification of putative miRNA targets and pathways in head and neck cancers. AB - MicroRNAs (miRNAs) are small noncoding RNAs that involved in various cancer related cellular processes. Diverse studies on expression profiling of miRNAs have been performed and the data showed that some miRNAs are up-regulated or down regulated in cancer. Until now, there are no data published on the miRNA expression in head and neck cancers from Malaysia. Hence, this study aimed to investigate potentially crucial miRNAs in head and neck cancer patients from Malaysian populations. A global miRNA profiling was performed on 12 samples of head and neck cancer tissue using microarray analysis followed by validation using real-time RT-PCR. Microarray analysis identified 10 miRNAs that could distinguish malignant head and neck cancer lesions from normal tissues; 7 miRNAs (hsa-miR-181a-2*, hsa-miR-29b-1*, hsa-miR-181a, hsa-miR-181b, hsa-miR-744, hsa miR-1271 and hsa-miR-221*) were up-regulated while 3 miRNAs (hsa-miR-141, hsa-miR 95 and hsa-miR-101) were down-regulated. These miRNAs may contribute in a simple profiling strategy to identify individuals at higher risk of developing head and neck cancers, thus helping in the elucidation of the molecular mechanisms involved in head and neck cancer pathogenesis. PMID- 21637913 TI - Expression and histopathological correlation of CCR9 and CCL25 in ovarian cancer. AB - Ovarian carcinoma is the most lethal gynecological malignancy among women and its poor prognosis is mainly due to metastasis. Chemokine receptor CCR9 is primarily expressed by a small subset of immune cells. The interactions between CCL25 and CCR9 have been implicated in leukocyte trafficking to the small bowel, a frequent metastatic site for ovarian cancer cells. We have previously shown that ovarian cancer cells express CCR9 and play an important role in cell migration, invasion and survival in the presence of its natural ligand in vitro. In this study, we have evaluated the expression of CCR9 and CCL25 in ovarian cancer cells and clinical samples. Ovarian cancer tissue microarrays from University of Alabama at Birmingham and AccuMax were stained for CCR9 and CCL25. Aperio ScanScope was used to acquire 80X digital images and expression analysis of CCR9 and CCL25. Flow cytometry and the Image stream system were used to conform the expression of CCR9 and CCL25 in ovarian cancer cells. Our results show significantly higher (p<0.001) expression of CCR9 and CCL25 in serous adenocarcinoma followed by serous papillary cystadenoma, endometrioid adenocarcinoma, mucinous adenocarcinoma, cystadenoma, mucinous boderline adenocarcinoma, clear cell carcinoma, granulosa cell tumor, dysgerminoma, transitional cell carcinoma, Brenner tumor, yolk sac tumor, adenocarcinoma and fibroma cases, compared to non neoplastic ovarian tissue. Similar to tissue expression, CCR9 was also significantly expressed by the ovarian cancer cell lines (OVCAR-3 and SK-OV-3) in comparison to normal adult ovarian epithelial cell. We provide the first evidence that CCR9 and its natural ligand CCL25 are highly expressed by ovarian cancer tissue and their expression correlates with histological subtypes. Expression of this chemokine receptor and its ligand CCL25 within primary tumor tissue further suggests a potential role of this chemokine-receptor axis in ovarian cancer progression. PMID- 21637914 TI - A novel GATA4 mutation responsible for congenital ventricular septal defects. AB - Ventricular septal defect (VSD) is the most common type of cardiovascular developmental anomaly and is an important risk factor for the substantially increased morbidity and mortality in newborns. Aggregating evidence implicates genetic defects in the pathogenesis of congenital VSD. However, VSD is genetically heterogeneous and the genetic determinants for VSD in most patients remain to be identified. In this study, the whole coding region of the GATA4 gene, which encodes a zinc-finger transcription factor pivotal to cardiogenesis, was initially sequenced in 210 unrelated patients with VSD. The relatives of the index patient carrying the identified mutation and 200 unrelated ethnically matched healthy individuals used as controls were subsequently genotyped. The functional effect of the mutant GATA4 was characterized in contrast to its wild type counterpart using a luciferase reporter assay system. A novel heterozygous GATA4 mutation, p.G296R, was identified in a family with VSD inherited as an autosomal dominant trait. Absent in 200 control individuals, the mutation co segregated with VSD in the family with 100% penetrance and was completely conserved evolutionarily across species. Functional analysis displayed that the p.G296R mutation of GATA4 was associated with a decreased transcriptional activity. The findings expand the spectrum of mutations in GATA4 linked to VSD and provide more insight into the molecular mechanism involved in VSD. The results of the present study imply the potential implications in the genetic diagnosis and gene-specific therapy of this common malformation in infancy. PMID- 21637915 TI - Synthetic emmprin peptides inhibit tumor cell-fibroblast interaction-stimulated upregulation of MMP-2 and tumor cell invasion. AB - Stromal cells are the main source of matrix metalloproteinases (MMPs) in human carcinoma tissues. Emmprin is a glycosylated transmembrane protein containing two immunoglobulin (Ig) domains that is expressed in carcinoma cells and stimulates MMP production by adjacent stromal cells. The first Ig domain (ECI) of emmprin contains the biologically active site. We investigated whether synthetic peptides carrying a partial ECI sequence could inhibit emmprin activity. Only the second peptide (emp#2), which contains a putative N-glycosylation site sequence, inhibited emmprin-stimulated production of MMP-2 in co-cultures of fibroblasts and several different human tumor cells types, including carcinoma, sarcoma, melanoma, leukemia and glioma cells. Moreover, emp#2 significantly inhibited the invasive activity of glioblastoma cells promoted by interaction with fibroblasts. Perturbation of emmprin activity by this peptide may have potential therapeutic uses in the prevention of MMP-2-dependent cancer invasion. PMID- 21637916 TI - 18alpha-glycyrrhetinic acid targets prostate cancer cells by down-regulating inflammation-related genes. AB - Glycyrrhetinic acid is an active triterpenoid metabolite of glycyrrhizin abundantly present in licorice roots. Glycyrrhetinic acid exists as alpha and beta stereo-isomeric forms. Both stereo-isomeric forms are known to have anti inflammatory and anticancer activity. However, the effects and anticancer mechanism of alpha glycyrrhetinic acid in prostate cancer cells has not yet been evaluated. Therefore, we investigated the growth inhibition, induction of apoptosis and the anticancer mechanisms of 18alpha-glycyrrhetinic acid (AGA), on the androgen-independent metastatic prostate cancer cell line DU-145. Our results showed that AGA inhibited proliferation and growth of these cells by inducing apoptosis as determined by Annexin V and flow cytometry analyses. Our studies also showed that HUVEC tube formation was drastically reduced when cultured in conditioned medium of AGA-treated DU-145 cells. In addition, AGA treatment prevented the invasion of DU-145 prostate cancer cells on matrigel coated transwells via down-regulation of NF-kappaB (p65), VEGF and MMP-9 expression. Furthermore, AGA treatment also down-regulated the expression of pro-inflammatory cytokine/growth factor genes HMGB1, IL-6 and IL-8 in DU-145 cells. Interestingly, AGA simultaneously upregulated the expression of non-steroidal anti-inflammatory gene-1 (NAG-1) in DU-145 cells suggesting its anti-inflammatory activity on prostate cancer cells. Taken together, the results of this study suggest that AGA may be a promising anticancer agent that merits further investigation for the chemoprevention and treatment of prostate cancer. PMID- 21637917 TI - BCR-ABL- and Ras-independent activation of Raf as a novel mechanism of Imatinib resistance in CML. AB - Although the BCR-ABL tyrosine kinase inhibitor Imatinib has undoubtedly revolutionized the therapy of chronic myeloid leukaemia (CML), acquired drug resistance remains a common problem in CML therapy. Resistance often arises from second-line mutations in BCR-ABL or overexpression of the BCR-ABL protein but in ~20% of CML cases resistance mechanisms do not involve altered BCR-ABL function. Imatinib-resistant CML cell lines have been widely used for comparative proteome/genome-wide expression screens in order to decipher resistance mechanisms but a clearcut molecular mechanism or molecular player in BCR-ABL independent resistance to Imatinib has not yet evolved from those studies. Here, we report the identification of a novel mechanism for Imatinib resistance in CML cells with unaltered BCR-ABL function. Pharmacological analysis evidenced a constitutive, Imatinib-insensitive activation of the Erk-MAPK pathway in resistant cells. A systematic analysis of pathway constituents illustrated that Ras-GTP accumulation remained fully sensitive to Imatinib but c-Raf activity from serum-fed cultures was largely resistant to the drug's action. Sequencing excluded mutations in either B-Raf or c-Raf as the origin of resistance, indicating that a functional alteration in the regulation of c-Raf activity was responsible for this effect. Collectively, these findings highlight a novel mechanism of acquired Imatinib resistance based on the BCR-ABL and Ras independent constitutive activation of the Erk-MAPK pathway through activated c Raf, which could prove helpful for a better functional classification of the causes of Imatinib resistance in CML. PMID- 21637918 TI - Identification of GIRK2-4 subunits in human esophageal smooth muscle cells. AB - Acetylcholine (ACh) secreted from the vagus nerve contributes to the physiological and pathological regulation of the contraction and relaxation of human esophageal smooth muscle. Expression of acetylcholine-sensitive G protein activated inwardly rectifying potassium channels (GIRKs) occurs widely in the heart, nervous system and gastrointestine, but the role of GIRKs in the esophagus remains unclear. In the present study, expression of the GIRK1-4 subunits in mRNA and total protein was examined in human esophageal smooth muscle cells (SMCs) by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. mRNA and protein expression of the GIRK2-4 subunits was detected in human esophageal longitudinal muscle (LM) and circular muscle (CM) cells. However, GIRK1 mRNA and protein were not observed in either the esophageal LM or CM. This study is the first to identify the expression of GIRK2-4 subunits in human esophageal SMCs. PMID- 21637919 TI - RB1CC1 activates the p16 promoter through the interaction with hSNF5. AB - RB1-inducible coiled-coil 1 (RB1CC1, also known as FIP200) is involved in dephosphorylation and increase of retinoblastoma tumor suppressor protein (RB1), but the RB1CC1 molecular mechanism in the dephosphorylation of RB1 is not fully understood. We determined that RB1CC1 activates the expression of p16 (also called INK4a/CDKN2a) through the activation of its promoter, using chromatin immunoprecipitation (ChIP) and p16 promoter-luciferase reporter assays. In addition, RB1CC1 essentially requires binding with hSNF5 (also known as BAF47/INI1, a chromatin-remodeling factor) to activate the p16 promoter, in order to enhance the RB1 pathway and acts as a tumor suppressor. Evaluation of the RB1CC1 mechanism of action is expected to provide useful information for clinical practice and future therapeutic strategies in human cancers. PMID- 21637920 TI - TTF1-induced apoptosis of HepG-2 cells through a mitochondrial pathway. AB - The Chinese medicinal herb Sorbaria sorbifolia, native to Changbai Mountain, can induce apoptosis in HepG-2 cells. We studied the mechanism by which 5, 2', 4' trihydroxy-6,7, 5'-trimethoxyflavone (TTF1) isolated from acetic ether extracts of Sorbaria sorbifolia induces apoptosis in HepG-2 cells. The results showed that TTF1 both inhibited cell growth and induced apoptosis. RT-PCR and Western blot analysis showed that TTF1 treatment led to decreased transcription and protein expression of bcl-2 and an increase in bax, Cyt-c, caspase-3 and caspase-9. These results together suggest that TTF1 may induce apoptosis of HepG-2 cells through a mitochondrial pathway. PMID- 21637921 TI - All reovirus subtypes show oncolytic potential in primary cells of human high grade glioma. AB - Reoviridae are non-human pathogenic viruses. The family of reoviridae consists of 4 different subtypes. Many studies have proven that the Dearing subtype 3 has oncolytic potential. This potential is related to the RAS protein expression in tumour cells. The aim of this study, was to investigate whether all reovirus subtypes have oncolytic potential and whether there are differences in their efficacy, in particular for high-grade glioma. To evaluate the oncolytic potential, we performed an in vitro head-to-head study for all reovirus subtypes in 5 primary cell cultures of high-grade gliomas. The oncolytic activity was determined using end-point titration with observation of the cytopathogenic effect. For measurement of RAS activity, we performed an immunofluorescent detection stain on all cell cultures. For quantification of the virus, an RT-PCR measurement for all subtypes was performed. All reovirus subtypes showed oncolytic activity in the observed glioma biopsies. These observations correlated with RAS overexpression in the observed cells. All glioma biopsies overexpressed the RAS protein. The quantitative oncolytic potential differed in relation to the single observed cell culture and in relation to the chosen reovirus subtype. To our knowledge, this is the first study showing oncolytic activity for all reovirus subtypes. We show the relationship and correlation between RAS protein overexpression and vulnerability of cells to reovirus. Efficacy of the different subtypes is interindividually different and cannot be forecast. PMID- 21637922 TI - Soy isoflavones augment the effect of TRAIL-mediated apoptotic death in prostate cancer cells. AB - Prostate cancer represents an ideal disease for chemopreventive intervention. Genistein, daidzein and equol, the predominant soy isoflavones, have been reported to lower the risk of prostate cancer. Isoflavones exert their chemopreventive properties by affecting apoptosis signalling pathways in cancer cells. Tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) is an endogenous anticancer agent that induces apoptosis selectively in tumour cells. Soluble or expressed in immune cells, TRAIL molecules play an important role in immune surveillance and defense mechanisms against tumour cells. However, various types of cancer cells are resistant to TRAIL-mediated apoptosis. We examined the cytotoxic and apoptotic effects of genistein, daidzein and equol in combination with TRAIL in LNCaP cells. Cytotoxicity was measured by MTT and LDH assays. Apoptosis was analyzed by flow cytometry and fluorescence microscopy using Annexin V-FITC. Mitochondrial membrane potential (DeltaPsim) was evaluated by fluorescence microscopy using DePsipher staining. Flow cytometry detected the expression of death receptor TRAIL-R1 (DR4) and TRAIL-R2 (DR5) on cell surfaces. The soy isoflavones sensitized TRAIL-resistant prostate cancer cells to apoptotic death. The isoflavones did not alter death receptor expression, but significantly augmented TRAIL-induced disruption of DeltaPsim in the LNCaP cells. We showed for the first time that the chemopreventive effects of soy foods on prostate cancer are associated with isoflavone-induced support of TRAIL-mediated apoptotic death. PMID- 21637923 TI - Down-regulation of beta-N-acetyl-D-glucosaminidase increases Akt1 activity in thyroid anaplastic cancer cells. AB - O-GlcNAcylation is a common and dynamic modification of intracellular proteins in which beta-N-acetyl-glucosamine moieties are attached to hydroxyl groups of serine or threonine residues (O-GlcNAc). Accumulating evidence suggests the critical role of protein O-GlcNAcylation in signal transduction, transcriptional control, cell cycle regulation and protein degradation. However, the exact role of O-GlcNAc modification in tumor pathogenesis or progression remains to be established. In the present study, we investigated the effect of increased O GlcNAcylation of cellular proteins on IGF-1 signaling in 8305C thyroid anaplastic cancer cells. The global O-GlcNAc level in the 8305C cells was increased by down regulation of beta-N-acetyl-D-glucosaminidase (O-GlcNAcase) activity, an enzyme which removes O-GlcNAc moieties. We demonstrated here that IGF-1 stimulates Akt1 activity in 8305C cells, and down-regulation of O-GlcNAcase activity by the chemical inhibitor PUGNAc or RNA interference method enhances this effect. Increased Akt1 activation increased cell proliferation. In cells with down regulation of O-GlcNAcase activity, kinase GSK3beta phosphorylation and cyclin D1 levels were higher than those in control cells. Our findings suggest that increased proliferation of 8305C cells treated with PUGNAc or RNAi against O GlcNAcase at least partially depends on the IGF-1-Akt1-GSK3beta-cyclin D1 pathway. PMID- 21637924 TI - Tetramethylpyrazine inhibits migration of SKOV3 human ovarian carcinoma cells and decreases the expression of interleukin-8 via the ERK1/2, p38 and AP-1 signaling pathways. AB - Interleukin-8 (IL-8) expression by melanoma cells may influence their metastatic capabilities. Tetramethylpyrazine (TMP) from Ligusticum wallichil Franch. possesses anti-inflammatory and antitumor activities. It has recently been suggested that autocrine IL-8 may play a role in tumor cell survival, invasion and migration. The role of TMP in association with IL-8 in the tumor cell migratory process remains unclear. The purpose of the present study was to determine whether TMP influences the migratory ability of a human ovarian carcinoma cell line (SKOV3) via regulation of IL-8 expression in vitro. Cell counts showed that treatment of SKOV3 with TMP (25-100 ug/ml) for 24 h did not decrease cell numbers, while an effect of TMP on the down-regulation of the expression of IL-8 was observed. In addition, migration of SKOV3 cells was suppressed after treatment with TMP (25-100 ug/ml) for 24 h. Therefore, expression of IL-8 by SKOV3 cells correlates with their metastatic potential. Western blot analysis revealed that ERK1/2 and p38 phosphorylation was blocked by TMP. Furthermore, IL-8 mRNA expression was inhibited significantly after co incubation with PD98059 (ERK inhibitor) and SB203580 (p38 inhibitor), respectively. Notably, these changes were the results of activator protein-1 (AP 1) activity suppression rather than that of NF-kappaB. Our data suggest that TMP may inhibit tumor cell invasion and migration, at least in part, through its down regulation of IL-8 expression. Our results provide evidence that anti inflammation plays an important role in integrative cancer therapies. PMID- 21637925 TI - Sepiapterin inhibits cell proliferation and migration of ovarian cancer cells via down-regulation of p70S6K-dependent VEGFR-2 expression. AB - Tetrahydrobiopterin (BH4) is known to be an essential cofactor for the aromatic amino acid hydroxylases, which are involved in the production of neurotransmitters, and for nitric oxide (NO) synthase. In the present study, we report that sepiapterin, the more stable form of the BH4 precursor, modulates ovarian cancer cell proliferation and migration by NO-dependent and -independent mechanisms. Sepiapterin induction of cell proliferation and migration in SKOV-3 cells is accompanied by ERK, Akt and p70S6K activation. These stimulatory effects of sepiapterin are reversed by pretreatment with NO synthase inhibitor. We also show that sepiapterin significantly inhibits vascular endothelial growth factor-A (VEGF-A)-stimulated cell proliferation and migration. Pretreatment with NO synthase inhibitor does not alter the ability of sepiapterin to inhibit VEGF-A induced cell proliferation and migration, indicating that the suppressive effects of sepiapterin on VEGF-A-induced responses are mediated by a NO-independent mechanism. Finally, we demonstrate that sepiapterin markedly suppresses VEGF-A induced p70S6K phosphorylation and VEGFR-2 expression, resulting in inhibition of VEGF-A-induced cell proliferation and migration. Collectively, these findings represent a biphasic effect of sepiapterin on cellular fates, depending on the presence of growth factors, and support further development and evaluation of sepiapterin for the treatment of cancers overexpressing VEGFR-2. PMID- 21637926 TI - Restored expression of the tumor suppressor gene RUNX3 reduces cancer stem cells in hepatocellular carcinoma by suppressing Jagged1-Notch signaling. AB - Runt-related transcription factor 3 (RUNX3) is a candidate tumor suppressor gene that is downregulated in various cancers. In the present study, we analyzed the regulatory function of RUNX3 on Jagged-1 (JAG1) expression and cancer stem cell (CSC) signaling in hepatocellular carcinoma (HCC). Eleven HCC cell lines and 30 human HCC tissues were used. RUNX3 and JAG1 expression levels were analyzed by immunoblotting and immunohistochemistry. Ectopic RUNX3 expression was induced by introducing RUNX3 cDNA into the RUNX3-negative HCC cell line Hep3B and Huh7 cells. Furthermore endogenous RUNX3 expression was knocked down by RUNX3 siRNA in SK-Hep-1 cells. In order to analyze JAG1 transcriptional regulation, we conducted reporter assays, chromatin immunoprecipitation (ChIP) assays and electrophoretic mobility shift assays (EMSAs). Tumorigenicity was analyzed using a SCID mouse liver injection model. An inverse correlation was observed between RUNX3 expression and JAG1 expression in most HCC cell lines and tissues. Restoring RUNX3 expression decreased the expression of JAG1 in Hep3B and Huh7 cells, whereas JAG1 expression was upregulated in RUNX3 siRNA-treated SK-Hep-1 cells. Reporter assays, ChIP assays and EMSAs revealed that RUNX3 directly bound to the transcriptional regulatory region of JAG1 and suppressed JAG1 transcription. Moreover, RUNX3 restoration downregulated CSCs by suppressing JAG1-mediated Notch signaling. The tumorigenic capacity of RUNX3-expressing Hep3B cells was lower compared to that of control Hep3B cells. RUNX3 expression suppressed JAG1 expression and resulted in downregulation of tumorigenesis by suppression of JAG1 mediated CSCs. PMID- 21637927 TI - Pyrolysis-GC/MS for the identification of macromolecular components in historical recipes. AB - Analytical pyrolysis with thermally assisted hydrolysis and methylation was employed to investigate ancient ointments collected from Spanish vessels coming from the sixteenth century pharmacies. The ointments were reproduced on the basis of historical recipes and characterization was made in comparison with real samples. Characteristic markers indicate the presence of beeswax, of animal and plant lipids, and of natural resins. Analyses of old samples are consistent with the modern reproductions and with the analysis of raw materials. Multivariate data analysis was used to discriminate between the different types of lipidic materials, also in connection with their relative amount in the samples. PMID- 21637928 TI - Bioaccumulation assessment of the sunscreen agent 2-ethylhexyl 4-(N,N dimethylamino)benzoate in human semen by automated online SPE-LC-MS/MS. AB - The proven endocrine disruption nature of the sunscreen ingredient 2-ethylhexyl 4 (N,N-dimethylamino)benzoate (EDP) calls for research to understand its distribution and bioaccumulation in the human body. A sensitive analytical method to determine EDP and its metabolites in human semen based on online SPE-LC-MS/MS is described. The method has been fully validated and a standard addition calibration has been used for quantification to correct the observed matrix effects. The on-column detection limits of the analytes are between 0.2 and 0.6 ng, depending on the analyte and the sample. The repeatability of the method, expressed as relative standard deviation, was in the range 4.6-9.4%. The method was satisfactorily applied to semen samples from male volunteers who were subjected to single and repeated whole-body applications of an EDP-containing sunscreen product. EDP metabolites were found at different concentrations in semen samples from the repeated application study, thus showing evidences of bioaccumulation in humans. PMID- 21637929 TI - Determination of some sulfonylurea herbicides in soil by a novel liquid-phase microextraction combined with sweeping micellar electrokinetic chromatography. AB - A novel method for the determination of five sulfonylurea herbicides in soil was developed by a dispersive solid-phase extraction (DSPE) clean-up followed by dispersive liquid-liquid microextraction (DLLME), prior to sweeping micellar electrokinetic chromatography (MEKC). In the DSPE-DLLME, 10 g of soil sample was first extracted with 10 mL of acetonitrile containing 5% formic acid (pH 3.0). The extract was then cleaned-up by a DSPE with C(18) as sorbent. A 1 mL aliquot of the resulting extract was then added into a centrifuge tube containing 5 mL of water adjusted to pH 2.0 and 60.0 MUL chlorobenzene (as extraction solvent) for DLLME procedure. Then, the organic sample extraction solution was evaporated to dryness, and reconstituted with 20.0 MUL of 1.0 mmol L(-1) Na(2)HPO(4) (pH 10.0) for sweeping-MEKC analysis after DLLME. Under optimized conditions, the method provided as high as 3,000- to 5,000-fold enrichments factors. The linearity of the method was in the range of 3.3-200 ng g(-1) for chlorimuron ethyl and bensulfuron methyl, and in the range of 1.7-200 ng g(-1) for tribenuron methyl, chlorsulfuron and metsulfuron methyl, with the correlation coefficients (r) ranging from 0.9965 to 0.9983, respectively. The limits of detection (LODs) ranged from 0.5 to 1.0 ng g(-1). The intraday relative standard deviations (RSDs, n = 5) were below 5.3% and interday RSDs (n = 15) within 6.8%. The recoveries of the method for the five sulfonylureas from soil samples at spiking levels of 5.0, 20.0, and 100.0 ng g(-1) were 76.0-93.5%, respectively. The developed method has been successfully applied to the analysis of the target sulfonylurea herbicide residues in soil samples with a satisfactory result. PMID- 21637930 TI - Quantitative multi-element mapping of ancient glass using a simple and robust LA ICP-MS rastering procedure in combination with image analysis. AB - The surface of two glass artefacts in mosaic style, probably fragments of conglomerate glass bowls dating back two millennia, was investigated by laser ablation-inductively coupled plasma mass spectrometry (LA-ICP-MS). By rastering with the laser beam over a selected area of the surface of the glass artefacts, elemental oxide maps were generated. Quantification of the elemental oxides in the maps was achieved using a so-called sum normalization procedure, summating the elements-54 in total-as their oxides to 100% (w/w), without using an internal standard and applying only one external standard (NIST SRM glass 610). This results in a robust mapping procedure which automatically corrects for drift and defocusing issues. Sum normalization was applied to each pixel in the map separately and required a custom source code to process all the data in the tens of thousands of pixels to generate the elemental oxide concentration maps. The digital element maps generated upon rastering of the two glass artefacts are very compelling and are an excellent entry point to gain detailed insight into their fabrication and provenance using image analysis software for retrieval of localized elemental oxide concentrations and correlations. PMID- 21637931 TI - Recent developments in doping testing for erythropoietin. AB - The constant development of new erythropoiesis-stimulating agents (ESAs), since the first introduction of recombinant erythropoietin (rhEpo) for clinical use, has also necessitated constant development of methods for detecting the abuse of these substances. Doping with ESAs is prohibited according to the World Anti Doping Code and its prohibited list of substances and methods. Since the first publication of a direct and urine-based detection method in 2000, which uses changes in the Epo isoform profile as detected by isoelectric focusing in polyacrylamide slab gels (IEF-PAGE), the method has been constantly adapted to the appearance of new ESAs (e.g., Dynepo, Mircera). Blood had to be introduced as an additional matrix, because Mircera (a PEGylated Epo) is best confirmed in serum or plasma after immunoaffinity purification. A Mircera ELISA was developed for fast screening of sera. With the appearance of Dynepo and copy epoetins, the additional application of sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE or equivalent) became necessary. The haematological module of the Athlete Biological Passport is the latest development in multivariable indirect testing for ESA doping. The article summarizes the main strategies currently used in Epo anti-doping testing with special focus on new developments made between 2009 and 2010. PMID- 21637932 TI - Discovering the composition of ancient cosmetics and remedies: analytical techniques and materials. AB - This article reviews the analytical techniques and procedures used in the study of ancient cosmetics, therapeutic chemicals, and remedies found in historical and archaeological sites. Well consolidated techniques based on molecular and atomic spectroscopy, for example FT-IR, Raman, SEM-EDX, and XRD, and analytical procedures based on high-performance chromatography and mass spectrometry, for example GC-MS and HPLC-MS are reviewed. The advantages of recently introduced techniques based on synchrotron radiation and on direct mass spectrometric techniques are also discussed. The possibility of extracting information about composition, preparation techniques, and the degradation processes of ancient cosmetics, pharmaceutics, and ritual balms is analysed by use of several case studies. PMID- 21637933 TI - Cannabinoids and metabolites in expectorated oral fluid after 8 days of controlled around-the-clock oral THC administration. AB - Oral fluid (OF) is an increasingly accepted matrix for drug testing programs, but questions remain about its usefulness for monitoring cannabinoids. Expectorated OF specimens (n = 360) were obtained from 10 adult daily cannabis smokers before, during, and after 37 20-mg oral Delta(9)-tetrahydrocannabinol (THC) doses over 9 days to characterize cannabinoid disposition in this matrix. Specimens were extracted and analyzed by gas chromatography-mass spectrometry with electron impact ionization for THC, 11-hydroxy-THC, cannabidiol, and cannabinol, and negative chemical ionization for 11-nor-9-carboxy-THC (THCCOOH). Linear ranges for THC, 11-hydroxy-THC, and cannabidiol were 0.25-50 ng/mL; cannabinol 1-50 ng/mL; and THCCOOH 5-500 pg/mL. THCCOOH was the most prevalent analyte in 344 specimens (96.9%), with concentrations up to 1,390.3 pg/mL. 11-hydroxy-THC, cannabidiol, and cannabinol were detected in 1, 1, and 3 specimens, respectively. THC was detected in only 13.8% of specimens. The highest THC concentrations were obtained at admission (median 1.4 ng/mL, range 0.3-113.6) from previously self administered smoked cannabis. A total of 2.5 and 3.7% of specimens were THC positive at the recommended Substance Abuse and Mental Health Services Administration (2 ng/mL) and Driving Under the Influence of Drugs, Alcohol and Medicines (DRUID) (1 ng/mL) confirmation cutoffs, respectively. THC is currently the only analyte for monitoring cannabis exposure in OF; however, these data indicate chronic therapeutic oral THC administration and illicit oral THC use are unlikely to be identified with current guidelines. Measurement of THCCOOH may improve the detection and interpretation of OF cannabinoid tests and minimize the possibility of OF contamination from passive inhalation of cannabis smoke. PMID- 21637934 TI - Toward poetic science. AB - This article is an extension of earlier discussion in the present journal regarding feelings in literature and, more broadly, the distinction between literary and scientific discourse. Valid though this distinction may be on some level, it is argued herein that it owes its very existence, in part, to a problematically narrow view of what constitutes science, one that not only eschews the life of feeling but that fails to abide by the primary obligation of the scientific endeavor-namely, to practice fidelity to the phenomena of concern. In this respect, it might plausibly be said that much of contemporary psychological science is not scientific enough and that, consequently, a more open and capacious conception is called for. More specifically, it is suggested that a portion of the discipline move in the direction of what might be called poetic science, a form of science that, in its very art-fulness, can do justice to the ambiguity, complexity, and depth that characterizes lived experience. PMID- 21637935 TI - Understanding continuity to recognize discontinuity. AB - In this paper the author comments on the contribution by Antonio P. Ribeiro and Miguel M. Goncalves (in this journal) that offer a creative and unique perspective on maintenance and transformation of problematic self-narrative. From here the author contributes to the topic through the exploration of some issues: a) the relation, in the dialogical process of self-narrative construction, between semiotic processes that give voice to the semantic opposition and semiotic processes that give voice to the contradictory; b) the relation between sameness and ipseity in the self-narrative process; c) the role of a pathemic axis of meaning in the generation process of self-narratives. A final reflection is done on narrative as a device of clinical intervention in which the author makes a distinction between methods based on the recognition and extension of variability and methods based on the recognition of permanency so to get to variability. PMID- 21637936 TI - PdCYP51B, a new putative sterol 14alpha-demethylase gene of Penicillium digitatum involved in resistance to imazalil and other fungicides inhibiting ergosterol synthesis. AB - Penicillium digitatum, causing green mold decay, is the most destructive postharvest pathogen of citrus fruits worldwide. The phenotypes and genotypes of 403 isolates of P. digitatum, collected from packing houses and supermarkets in Zhejiang, China, during 2000 to 2010, were characterized in terms of their imazalil sensitivity. The frequency of detected imazalil-resistant (IMZ-R) isolates increased from 2.1% in 2000 to 60-84% during 2005-2010. Only 6.5% and 4.5% of the collected IMZ-R isolates belong to the previously described IMZ-R1 and IMZ-R2 genotypes, respectively. To determine the resistance mechanism of the predominant and novel IMZ-R isolates of P. digitatum (termed IMZ-R3), genes PdCYP51B and PdCYP51C, homologous to the sterol 14alpha-demethylase encoded gene PdCYP51, were cloned from six IMZ-R3 and eight imazalil-sensitive (IMZ-S) isolates of P. digitatum. A unique 199-bp insertion was observed in the promoter region of PdCYP51B in all IMZ-R3 isolates examined but in none of the tested IMZ S isolates. Further analysis by PCR confirmed that this insertion was present in all IMZ-R3 isolates but absent in IMZ-S, IMZ-R1, and IMZ-R2 isolates. Transcription levels of PdCYP51B in three IMZ-R3 isolates were found to be 7.5- to 13.6-fold higher than that in two IMZ-S isolates of P. digitatum. Introduction of another copy of PdCYP51B ( s ) (from IMZ-S) into an IMZ-S isolate decreased the sensitivity of P. digitatum to 14alpha-demethylation inhibitors (DMIs) only to a small extent, but introduction of a copy of PdCYP51B ( R ) (from IMZ-R3) dramatically increased the resistance level of P. digitatum to DMIs. Regarding PdCYP51C, no consistent changes in either nucleotide sequence or expression level were correlated with imazalil resistance among IMZ-R and IMZ-S isolates. Based on these results, we concluded that (1) the CYP51 family of P. digitatum contains the PdCYP51B and PdCYP51C genes, in addition to the known gene PdCYP51A (previously PdCYP51); (2) PdCYP51B is involved in DMI fungicide resistance; and (3) overexpression of PdCYP51B resulting from a 199-bp insertion mutation in the promoter region of PdCYP51B is responsible for the IMZ-R3 type of DMI resistance in P. digitatum. PMID- 21637937 TI - Prokaryotic diversity, composition structure, and phylogenetic analysis of microbial communities in leachate sediment ecosystems. AB - In order to obtain insight into the prokaryotic diversity and community in leachate sediment, a culture-independent DNA-based molecular phylogenetic approach was performed with archaeal and bacterial 16S rRNA gene clone libraries derived from leachate sediment of an aged landfill. A total of 59 archaeal and 283 bacterial rDNA phylotypes were identified in 425 archaeal and 375 bacterial analyzed clones. All archaeal clones distributed within two archaeal phyla of the Euryarchaeota and Crenarchaeota, and well-defined methanogen lineages, especially Methanosaeta spp., are the most numerically dominant species of the archaeal community. Phylogenetic analysis of the bacterial library revealed a variety of pollutant-degrading and biotransforming microorganisms, including 18 distinct phyla. A substantial fraction of bacterial clones showed low levels of similarity with any previously documented sequences and thus might be taxonomically new. Chemical characteristics and phylogenetic inferences indicated that (1) ammonium utilizing bacteria might form consortia to alleviate or avoid the negative influence of high ammonium concentration on other microorganisms, and (2) members of the Crenarchaeota found in the sediment might be involved in ammonium oxidation. This study is the first to report the composition of the microbial assemblages and phylogenetic characteristics of prokaryotic populations extant in leachate sediment. Additional work on microbial activity and contaminant biodegradation remains to be explored. PMID- 21637938 TI - A sirA-like gene, sirA2, is essential for 3-succinoyl-pyridine metabolism in the newly isolated nicotine-degrading Pseudomonas sp. HZN6 strain. AB - A novel nicotine-degrading Pseudomonas sp. strain, HZN6, was isolated from a pesticide-wastewater treatment facility in Hangzhou. The strain could grow on nicotine as its sole source of carbon, nitrogen, and energy. The strain's main intermediate metabolites were determined to be pseudooxynicotine, 3-succinoyl pyridine (SP), and 6-hydroxy-3-succinoyl-pyridine (HSP). A Tn5 transposon mutant was generated in which the degradation pathway was blocked at the SP. A 4,583-bp DNA fragment flanking the transposon insertion site was obtained through self formed adaptor PCR and analyzed. The mutant gene orfC displays 89% deduced amino acid sequence identity with the sirA-like gene (sirA2, a sulfurtransferase homologue gene) of Pseudomonas stutzeri A1501. The orfC-disrupted strain lost the ability to degrade SP, and the complementation strains with the orfC from the Pseudomonas sp. HZN6 and the sirA2 (PP_1233) from Pseudomonas putida KT2440 recovered the degradation ability. Though the orfC-disrupted strain also lost the xanthine dehydrogenase activity, the effects of tungsten on the degradation of SP and hypoxanthine revealed that the hydroxylation of SP to HSP was not a xanthine dehydrogenase type. These results demonstrated that the orfC gene was essential for the SP metabolism involved in the nicotine metabolic pathway in the Pseudomonas sp. HZN6 strain. This study might advance the understanding of the nicotine metabolic mechanism in Pseudomonas. PMID- 21637939 TI - Identification of a cyclooxygenase gene from the red alga Gracilaria vermiculophylla and bioconversion of arachidonic acid to PGF(2alpha) in engineered Escherichia coli. AB - Prostaglandins (PGs) are important local messenger molecules in many tissues and organs of animals including human. For applications in medicine and animal care, PGs are mostly purified from animal tissues or chemically synthesized. To generate a clean, reliable, and inexpensive source for PGs, we have now engineered expression of a suitable cyclooxygenase gene in Escherichia coli and achieved production levels of up to 2.7 mg l(-1) PGF(2alpha). The cyclooxygenase gene cloned from the red alga Gracilaria vermiculophylla appears to be fully functional without any eukaryotic modifications in E. coli. A crude extract of the recombinant E. coli cells is able to convert in vitro the substrate arachidonic acid (AA) to PGF(2alpha). Furthermore, these E. coli cells produced PGF(2alpha) in a medium supplemented with AA and secreted the PGF(2alpha) product. To our knowledge, this is the first report of the functional expression of a cyclooxygenase gene and concomitant production of PGF(2alpha) in E. coli. The successful microbial synthesis of PGs with reliable yields promises a novel pharmaceutical tool to produce PGF(2alpha) at significantly reduced prices and greater purity. PMID- 21637940 TI - Improved homo L-lactic acid fermentation from xylose by abolishment of the phosphoketolase pathway and enhancement of the pentose phosphate pathway in genetically modified xylose-assimilating Lactococcus lactis. AB - In order to achieve efficient homo L-lactic acid fermentation from xylose, we first carried out addition of xylose assimilation ability to Lactococcus lactis IL 1403 by introducing a plasmid carrying the xylRAB genes from L. lactis IO-1 (pXylRAB). Then modification of xylose assimilation pathway was carried out. L. lactis has two pathways for xylose assimilation called the phosphoketolase pathway (PK pathway) that produces both lactic acid and acetic acid and the pentose phosphate pathway (PP pathway) that produces only lactic acid as a final product. Thus a mutant strain that disrupted its phosphokeolase gene (ptk) was constructed. The Deltaptk mutant harboring pXylRAB lacked the PK pathway and produced predominantly lactic acid from xylose via the PP pathway, although its fermentation rate slightly decreased. Further introduction of the transketolase gene (tkt) to disrupted ptk locus led restoration of fermentation rate and this was attributed to enhancement of the PP pathway. As a result, ptk::tkt strain harboring pXylRAB produced 50.1 g/l of L-lactic acid from xylose with a high optical purity of 99.6% and a high yield of 1.58 (moles per mole xylose consumed) that is close to theoretical value of 1.67 from xylose. PMID- 21637941 TI - Improved EM algorithm for MPT model analysis. AB - Multinomialprocessing tree (MPT) models are in wide use as measurement models for analyzing categorical data in cognitive experiments. The approach involves estimating parameters and conducting hypothesis tests involving parameters that are arrayed in a tree structure designed to represent latent cognitive processes. The standard inference algorithm for these models is based on the well-known expectationmaximization (EM) algorithm. On the basis of the original use of the EMalgorithm for MPT models, this article presents an approach that accelerates the convergence speed of the algorithm by (1) adjusting suitable initial positions for certain parameters to reduce required iterative times and (2) using a series of operations between/among a set of matrices that are specific to the original model structure and information to reduce the time required for a single iteration. As compared with traditional algorithms, the simulation results show that the proposed algorithm has superior efficiency in interpreted languages and also has better algorithm readability and structure flexibility. PMID- 21637942 TI - Content-embedded tasks beat complex span for predicting comprehension. AB - Discourse comprehension requires one to process information that is actively maintained in working memory (WM). Therefore, we hypothesized that individual differences in comprehension would be predicted better by working memory tasks that capture the concurrent demands of processing and maintenance of the same memory elements (i.e., content-embedded tasks) than by WM tasks that require the maintenance of an extraneous memory load during processing (e.g., complex span tasks). Two hundred sixty-one undergraduates completed three content-embedded tasks, three complex span tasks, and three measures of comprehension. Results of structural equation modeling indicated that the content-embedded tasks accounted for a greater amount of variance in comprehension than did complex span tasks. Thus, tasks that require one to coordinate the processing and maintenance of task specific memory elements are preferable for capturing the relationship between WM and comprehension. PMID- 21637943 TI - Systematic influence of gaze position on pupil size measurement: analysis and correction. AB - Cognitive effort is reflected in pupil dilation, but the assessment of pupil size is potentially susceptible to changes in gaze position. This study exemplarily used sentence reading as a stand-in for paradigms that assess pupil size in tasks during which changes in gaze position are unavoidable. The influence of gaze position on pupil size was first investigated by an artificial eye model with a fixed pupil size. Despite its fixed pupil size, the systematic measurements of the artificial eye model revealed substantial gaze-position-dependent changes in the measured pupil size. We evaluated two functions and showed that they can accurately capture and correct the gaze-dependent measurement error of pupil size recorded during a sentence-reading and an effortless z-string-scanning task. Implications for previous studies are discussed, and recommendations for future studies are provided. PMID- 21637944 TI - Enhanced solubility, stability, and transcorneal permeability of delta-8 tetrahydrocannabinol in the presence of cyclodextrins. AB - The purpose of this study was to investigate the effect of cyclodextrins (CDs) on aqueous solubility, stability, and in vitro corneal permeability of delta-8 tetrahydrocannabinol (Delta(8)-THC). Phase solubility of Delta(8)-THC was studied in the presence of 2-hydroxypropyl-beta-cyclodextrin (HPbetaCD), randomly methylated-beta-cyclodextrin (RMbetaCD) and sulfobutyl ether-beta-cyclodextrin sodium salt (SbetaCD). Stability of Delta(8)-THC in 5% w/v aqueous CD solutions, as a function of pH, was studied following standard protocols. In vitro corneal permeation of Delta(8)-THC (with and without CDs) across excised rabbit cornea was also determined. Phase-solubility profile of Delta(8)-THC in the presence of both HPbetaCD and RMbetaCD was of the A(P) type, whereas, with SbetaCD an A(L) type was apparent. Aqueous solubility of Delta(8)-THC increased to 1.65, 2.4, and 0.64 mg/mL in the presence of 25% w/v HPbetaCD, RMbetaCD, and SbetaCD, respectively. Significant degradation of Delta(8)-THC was not observed within the study period at the pH values studied, except for at pH 1.2. Transcorneal permeation of Delta(8)-THC was dramatically improved in the presence of CDs. The results demonstrate that CDs significantly increase aqueous solubility, stability, and transcorneal permeation of Delta(8)-THC. Thus, topical ophthalmic formulations containing Delta(8)-THC and modified beta CDs may show markedly improved ocular bioavailability. PMID- 21637945 TI - Development of lipid-based nanoparticles for enhancing the oral bioavailability of paclitaxel. AB - The current research work investigates the potential of solid lipid nanoparticles (SLNs) in improving the oral bioavailability of paclitaxel. Paclitaxel-loaded SLNs (PTX-SLNs) were prepared by modified solvent injection method using stearylamine as lipid, soya lecithin and poloxamer 188 as emulsifiers. SLNs were characterized in terms of surface morphology, size and size distribution, surface chemistry and encapsulation efficiency. Pharmacokinetics and bioavailability studies were conducted in male Swiss albino mice after oral administration of PTX SLNs. SLNs exhibited spherical shape with smooth surface as analyzed by transmission electron microscopy (TEM). The mean particle size of SLNs was 96 +/- 4.4 nm with a low polydispersity index of 0.162 +/- 0.04 and zeta potential of 39.1 +/- 0.8 mV. The drug entrapment efficiency was found to be 75.42 +/- 1.5% with a loading capacity of 31.5 +/- 2.1% (w/w). Paclitaxel showed a slow and sustained in vitro release profile and followed Higuchi kinetic equations. After oral administration of the PTX-SLNs, drug exposure in plasma and tissues was ten- and twofold higher, respectively, when compared with free paclitaxel solution. PTX-SLNs produced a high mean C (max) (10,274 ng/ml) compared with that of free paclitaxel solution (3,087 ng/ml). The absorbed drug was found to be distributed in liver, lungs, kidneys, spleen, and brain. The results suggested that PTX-SLNs dispersed in an aqueous environment are promising novel formulations that enhanced the oral bioavailability of hydrophobic drugs, like paclitaxel and were quite safe for oral delivery of paclitaxel as observed by in vivo toxicity studies. PMID- 21637946 TI - Bioavailability study of berberine and the enhancing effects of TPGS on intestinal absorption in rats. AB - Berberine chloride (BBR) is a natural isoquinoline alkaloid extracted from medicinal herbs. It has been reported that the intestinal absorption of BBR is very low. In this study, the absolute bioavailability of BBR was studied, and the enhancing effects of D-alpha-tocopheryl polyethylene glycol 1000 succinate (TPGS) on intestinal absorption were investigated in rats. BBR injection was administrated via the femoral vein at a dose of 1.0 mg kg(-1) in intravenous group, and BBR oral formulations were administrated by oral gavage at a dose of 100 mg kg(-1) in BBR control (control) group and BBR-TPGS (test) group, respectively. The result showed that BBR had a very low absolute bioavailability of 0.68%, and TPGS could enhance intestinal absorption of BBR significantly. TPGS at a concentration of 2.5% could improve peak concentration (C(max)) and area under the curve (AUC(0-36)) of BBR by 2.9 and 1.9 times, respectively. The absorption enhancing ability of TPGS may be due to its ability to affect the biological activity of P-glycoprotein and thereby reduce the excretion of absorbed BBR into the intestinal lumen. This study indicated that absolute bioavailability of BBR was 0.68% in rats, and TPGS was a good absorption enhancer capable of enhancing intestinal absorption of BBR significantly. PMID- 21637949 TI - Sleep disturbances in untreated Parkinson's disease. AB - Sleep abnormalities are frequently found in Parkinson's disease (PD). However, it is unclear if they are present from the initial stages of PD. We thus aimed to assess sleep disturbances in newly diagnosed PD patients. We investigated 20 untreated PD patients using the Epworth Sleepiness Scale (ESS), the Pittsburgh Sleep Quality Index (PSQI) and the PD Sleep Scale (PDSS). Video-polysomnography and multiple sleep latency test (MSLT) were performed in 15 patients and 15 healthy controls. The ESS score was abnormally high in one patient, while short MSLT times were found in three other patients. The PSQI was higher (p < 0.05) and the PDSS lower (p < 0.001) in patients compared with controls. Video polysomnography demonstrated a higher percentage of rapid eye movement sleep without atonia (RWA) in patients compared with controls (mean 28 vs. 2.9%, p < 0.001), whereas only one patient had clinically manifested rapid eye movement sleep behavior disorder (RBD). Interestingly, the occurrence of RWA correlated with the motor score (rho = 0.65, p < 0.05). This study demonstrates that sleep disturbances emerge, in a proportion of patients, from the early stages of PD. RWA is a common finding while RBD is rarely present in early untreated PD. PMID- 21637948 TI - Vimentin in cancer and its potential as a molecular target for cancer therapy. AB - Vimentin, a major constituent of the intermediate filament family of proteins, is ubiquitously expressed in normal mesenchymal cells and is known to maintain cellular integrity and provide resistance against stress. Vimentin is overexpressed in various epithelial cancers, including prostate cancer, gastrointestinal tumors, tumors of the central nervous system, breast cancer, malignant melanoma, and lung cancer. Vimentin's overexpression in cancer correlates well with accelerated tumor growth, invasion, and poor prognosis; however, the role of vimentin in cancer progression remains obscure. In recent years, vimentin has been recognized as a marker for epithelial-mesenchymal transition (EMT). Although EMT is associated with several tumorigenic events, vimentin's role in the underlying events mediating these processes remains unknown. By virtue of its overexpression in cancer and its association with tumor growth and metastasis, vimentin serves as an attractive potential target for cancer therapy; however, more research would be crucial to evaluate its specific role in cancer. Our recent discovery of a vimentin-binding mini-peptide has generated further impetus for vimentin-targeted tumor-specific therapy. Furthermore, research directed toward elucidating the role of vimentin in various signaling pathways would reveal new approaches for the development of therapeutic agents. This review summarizes the expression and functions of vimentin in various types of cancer and suggests some directions toward future cancer therapy utilizing vimentin as a potential molecular target. PMID- 21637950 TI - Treatment of multiple sclerosis: current concepts and future perspectives. AB - Currently, several disease-modifying therapies for the treatment of multiple sclerosis are established and more are likely to be introduced. These treatment options differ with respect to their application profile, mechanism of action, efficacy, safety, and tolerance. Here, we review current concepts of MS therapies, report recent results of clinical trials, and discuss emerging treatment options. PMID- 21637951 TI - A common variant in the adiponectin gene and polycystic ovary syndrome risk. AB - In this study, we explored whether polymorphisms in insulin receptor (INSR), adiponectin (ADIPOQ), parathyroid hormone (PTH), and vitamin D receptor (VDR) genes are associated with polycystic ovary syndrome (PCOS). A total of 362 subjects, including 181 women with PCOS and 181 controls were enrolled in this case-control study. Two SNPs (rs2059806 and rs1799817) in the INSR gene, two SNPs (rs2241766 and rs1501299) in the ADIPOQ gene, one SNP (rs6256) in the PTH gene, and one SNP (rs757343) in the VDR gene were analyzed using PCR-RFLP method. We observed no significant difference in genotype and allele frequencies between the women with PCOS and controls for the rs2059806, rs1799817, rs1501299, rs6256, and rs757343 polymorphisms either before or after adjustment for confounding factors including age and BMI. However, the ADIPOQ rs2241766 "TT" genotype compared with "TG and GG" genotypes was associated with a 1.93-fold increased risk for PCOS (P = 0.006, OR = 1.93, 95% CI = 1.20-3.11), and the differences remained significant after adjustment for age and BMI (P = 0.039, OR = 1.72, 95% CI = 1.03-2.86). Furthermore, the ADIPOQ rs2241766 "T" allele was significantly overrepresented in women with PCOS than controls (P = 0.006; OR = 1.80, 95% CI = 1.18-2.70), and the difference remained significant after Bonferroni correction. Our findings suggest that the ADIPOQ rs2241766 "TT" genotype is a marker of increased PCOS susceptibility. This study also indicates for the first time that there are no significant association between INSR rs2059806, PTH rs6256, and VDR rs757343 gene polymorphisms and PCOS risk. However, these data remain to be confirmed in larger studies and in other populations. PMID- 21637952 TI - Plk1-mediated phosphorylation of UAP56 regulates the stability of UAP56. AB - Polo-like kinase 1 (Plk1) is a conserved serine/threonine protein kinase that plays pivotal roles during the cell cycle and cell proliferation. Although a number of important targets have been identified, the mechanism of Plk1-regulated pathways and the bulk of the Plk1 interactome are largely unknown. Here, we demonstrate that Plk1 interacts with the DExH/D RNA helicase, UAP56. The protein levels of UAP56 and Plk1 are inversely correlated during the cell cycle. We also show that Plk1 phosphorylates UAP56 in vitro and in vivo and that Plk1-dependent phosphorylation of UAP56 triggers ubiquitination and degradation of UAP56 through proteasomes. This result suggests that Plk1-mediated phosphorylation of UAP56 regulates the stability of UAP56. Our results will be helpful in further understanding mRNA metabolism, cell cycle progression, and the link between mRNA metabolism and cellular function. PMID- 21637953 TI - Cytokine gene polymorphism and asthma susceptibility, progress and control level. AB - Asthma is a multifactor inflammatory disorder, and its management requires understanding of its various pathogenesis and control mechanisms. Cytokines and other inflammatory mediators are important factors in asthma pathophysiology. In this study, we evaluated the role of cytokine polymorphisms in the asthma susceptibility, progress, control, and lung functions. IL-4-C590T polymorphism by PCR-RFLP method, IFN-gamma T+874A, TNF-alpha-A308G, IL-6 G-174C and TGF-beta T+869C variants by ARMS-PCR method and IgE serum level by ELISA technique were determined in 81 asthmatic patients and 124 normal subjects. Asthma diagnosis, treatment and control levels were considered using standard schemes and criteria. TNF-alpha-308GA genotype was more frequent in asthmatics (P = 0.025, OR 3.352), and polymorphisms between different asthma control levels (P > 0.05) were not different. IFN-gamma+874AT genotype had a positive correlation with the familial history of asthma (P = 0.034, OR 2.688). IL-6-174C allele (P = 0.045), TNF-alpha 308GG genotype (P = 0.002) and TNF-alpha-308G allele (P = 0.004) showed reduced values, and TNF-alpha-308GA genotype (P = 0.002) increased FEF25-75 value in asthmatics. IFN-gamma+874AA genotype caused a decrease in FVC factor (P = 0.045). This study showed that TNF-alpha-308GA is a risk factor for asthma, but cytokine gene variants do not affect asthma control and IgE serum levels. Variants producing lower levels of IL-6, TNF-alpha and IFN-gamma are associated with reduced pulmonary capacities. To achieve an appropriate schema for asthma management, further studies with consideration of different aspects in a larger group of patients would be more elucidative. PMID- 21637954 TI - Characterization of the expression profiles of calpastatin (CAST) gene in chicken. AB - The calpain system, a Ca(2+)-activated protease family, plays an important role in postmortem tenderization of skeletal muscle due to its involvement in the degradation of important myofibrillar and associated proteins, as well as in cytoskeletal remodeling and regulation of muscle growth. In this study, we quantified the expression of calpastatin (CAST) in two Chinese chicken breeds (mountainous black-bone chicken breed (MB) and a commercial meat type chicken breed (S01)), to discern the tissue and age-related specific expression pattern and its potential role on muscle tissue metabolism. Real-time quantitative PCR (RT-qPCR) assay was developed for accurate measurement of CAST mRNA levels in various tissues from chicken with different ages (0, 2, 4, 6, 8, 10, and 12 week). CAST mRNA was detected in collected organs. The heart and leg muscle tissues had the highest expression of CAST than other tissues from the same chicken (P < 0.01). Age-related expression pattern of CAST gene was evident in breast muscle, liver, and brain tissues (P < 0.05), but not in heart and leg muscle tissues (P > 0.05). Overall, the CAST mRNA level exhibited a "rise-decline rise-decline" developmental change in breast muscle and liver, with the highest expression at 2 weeks and the lowest expression at 8 weeks. The S01 chicken had significantly higher expression of CAST in breast muscle and heart than the MB chicken (P < 0.05) at 10 weeks. Our results suggested the CAST expression may be related to muscle fiber development. PMID- 21637955 TI - Aldose reductase regulates TNF-alpha-induced inducible nitric oxide synthase expression in human mesangial cells. AB - Glomerulonephritis is one of the most important causes of renal failure, which is accompanied with production of Nitric Oxide (NO) synthesized by inducible nitric oxide synthase (iNOS). Aldose reductase (AR) is the key enzyme in polyol pathway and plays an important role in glucose metabolism. Here, we report our finding that AR regulates tumor-necrosis-factor-alpha-induced (TNF-alpha-induced) iNOS expression in human mesangial cells (HMC) via nuclear factor kappaB (NFkappaB) signal pathway. The TNF-alpha-induced iNOS expression in HMC with different level of AR was measured by Real-time PCR and Western blot. The activation of signal pathway was analyzed by Western blot and electrophoretic mobility shift assay (EMSA). In cultured HMC, TNF-alpha induces the expression of iNOS, which is attenuated by knockdown AR with siRNA. On the other side, transfection with pcDNA3.1-AR gets the consistent conclusion. Furthermore, knockdown of AR attenuated activity of NFkappaB, suggesting that the effects of AR on this pathway may result in the reduced iNOS transcription and expression. Taken together, these data demonstrate the role of AR in regulating iNOS expression induced by TNF-alpha in cultured HMC, indicating the novel function of AR in glomerulonephritis besides glucose metabolism. PMID- 21637956 TI - Mitochondrial DNA variation in natural populations of endangered Indian feather back fish, Chitala chitala. AB - Genetic variation at mitochondrial cytochrome b (cyt b) and D-loop region reveals the evidence of population sub-structuring in Indian populations of highly endangered primitive feather-back fish Chitala chitala. Samples collected through commercial catches from eight riverine populations from different geographical locations of India were analyzed for cyt b region (307 bp) and D-loop region (636 716 bp). The sequences of the both the mitochondrial regions revealed high haplotype diversity and low nucleotide diversity. The patterns of genetic diversity, haplotypes networks clearly indicated two distinct mitochondrial lineages and mismatch distribution strongly suggest a historical influence on the genetic structure of C. chitala populations. The baseline information on genetic variation and the evidence of population sub-structuring generated from this study would be useful for planning effective strategies for conservation and rehabilitation of this highly endangered species. PMID- 21637957 TI - Identification and expression pattern of one stress-responsive NAC gene from Solanum lycopersicum. AB - NAC (for NAM, ATAF1, 2, and CUC2) family genes have been found to play an important role in diversified developmental processes and environmental responses. A new NAC-type transcription factor SlNAC3 was primarily identified and isolated from the cDNA libraries of tomato cultivar Ailsa Craig. It contains three exons and two introns within genomic DNA sequence and encodes a polypeptide of 329 amino acids. A plant-specific and conserved NAC domain is located in the N terminus of SlNAC3. The protein SlNAC3 is subcellularly localized in the nucleus of onion epidemical cells and it has a transcriptional activation domain in the C terminal region which shows extremely divergent among NACs. Phylogenetic analysis showed that SlNAC3 belonged to the OsNAC3 subgroup of the NAC protein family. Tissue expression profile analysis revealed that SlNAC3 was expressed mainly in flower, fruit and root. The transcription expression of SlNAC3 was inhibited by salt, drought stress and ABA treatment. These data demonstrate that SlNAC3 might interact with environmental and endogenous stimuli and probably function when plants response to salt and drought stresses through ABA signaling pathways as a transcriptional activator. PMID- 21637958 TI - Hemi-Castaing ligamentoplasty for the treatment of chronic lateral ankle instability: a retrospective assessment of outcome. AB - PURPOSE: In the treatment of chronic ankle instability, most non-anatomical reconstructions use the peroneus brevis tendon. This, however, sacrifices the natural ankle stabilising properties of the peroneus brevis muscle. The aim of this study was to evaluate the functional outcome of patients treated with a hemi Castaing procedure, which uses only half the peroneus brevis tendon. METHODS: We performed a retrospective cohort study of patients who underwent hemi-Castaing ligamentoplasty for chronic lateral ankle instability between 1993 and 2010, with a minimum of one year follow-up. Patients were sent a postal questionnaire comprising five validated outcome measures: Olerud-Molander Ankle Score (OMAS), Karlsson Ankle Functional Score (KAFS), Tegner Activity Level Score (pre-injury, prior to surgery, at follow-up), visual analog scale on pain (VAS) and the Short Form 36 (SF-36). RESULTS: Twenty patients completed the questionnaire on functional outcome. The OMAS showed good to excellent outcome in 80% and the KAFS in 65%, the Tegner Score improved from surgery but did not reach pre-injury levels, the VAS on pain was 1 of 10 and the SF-36 returned to normal compared with the average population. CONCLUSIONS: Even though most patients were satisfied with the results, outcome at long-term follow-up was less favourable compared with the literature on anatomical reconstructions. In accordance with the literature, we therefore conclude that the initial surgical treatment of chronic lateral ankle instability should be an anatomical repair with augmentation (i.e. the Brostrom-Gould technique) and the non-anatomical repair should be reserved for unsuccessful cases after anatomical repair or in cases where no adequate ligament remnants are available for reconstruction. PMID- 21637959 TI - Percutaneous vertebroplasty versus balloon kyphoplasty for treatment of osteoporotic vertebral compression fracture: a meta-analysis of randomised and non-randomised controlled trials. AB - PURPOSE: There is still debate over whether vertebroplasty (VP) or kyphoplasty (KP) is superior for the treatment of osteoporosis vertebral compression fractures (VCFs). We performed a systematic review and meta-analysis of randomised and non-randomised controlled trials comparing VP with KP to reach a relatively conclusive answer. METHODS: We searched computerised databases comparing efficacy and safety of VP and KP in osteoporotic fractures. These trials reported pain relief (Visual Analogue Scale), disability (Oswestry disability score) and complications (i.e., cement leakage, incident fractures) as the primary outcome. RESULTS: Eight studies involving 848 patients were identified. The outcome showed that VP is more effective in the short-term (no more than seven days) pain relief. Kyphoplasty had a superior capability for intermediate-term (around three months) functional improvement. As for long-term pain relief and functional improvement, there is no significant difference between these two interventions. Consistently, both interventions have similar risk for subsequent fracture and cement leakage. CONCLUSION: Thus considering the higher cost of the KP procedure, we recommend VP over KP for the treatment of osteoporotic VCFs. PMID- 21637960 TI - Cardiac tamponade due to perforation by an Amplatzer atrial septal occluder in a patient with Marfan syndrome. AB - Device closure of atrial septum defect was performed using an Amplatzer septal occluder in a 48-year-old patient with Marfan syndrome. Acute tamponade due to perforation was observed 2 months after catheter intervention. Careful consideration of the indication for device closure for atrium septal defect is necessary in patients with Marfan syndrome. PMID- 21637961 TI - A structural view of the antibiotic degradation enzyme NDM-1 from a superbug. AB - Gram-negative Enterobacteriaceae with resistance to carbapenem conferred by New Delhi metallo-beta-lactamase 1 (NDM-1) are a type of newly discovered antibioticresistant bacteria. The rapid pandemic spread of NDM-1 bacteria worldwide (spreading to India, Pakistan, Europe, America, and Chinese Taiwan) in less than 2 months characterizes these microbes as a potentially major global health problem. The drug resistance of NDM-1 bacteria is largely due to plasmids containing the blaNDM-1 gene shuttling through bacterial populations. The NDM-1 enzyme encoded by the blaNDM-1 gene hydrolyzes beta-lactam antibiotics, allowing the bacteria to escape the action of antibiotics. Although the biological functions and structural features of NDM-1 have been proposed according to results from functional and structural investigation of its homologues, the precise molecular characteristics and mechanism of action of NDM-1 have not been clarified. Here, we report the three-dimensional structure of NDM-1 with two catalytic zinc ions in its active site. Biological and mass spectroscopy results revealed that D-captopril can effectively inhibit the enzymatic activity of NDM-1 by binding to its active site with high binding affinity. The unique features concerning the primary sequence and structural conformation of the active site distinguish NDM-1 from other reported metallo-beta-lactamases (MBLs) and implicate its role in wide spectrum drug resistance. We also discuss the molecular mechanism of NDM-1 action and its essential role in the pandemic of drug-resistant NDM-1 bacteria. Our results will provide helpful information for future drug discovery targeting drug resistance caused by NDM-1 and related metallo-beta-lactamases. PMID- 21637962 TI - Targeted deletion of mouse Rad1 leads to deficient cellular DNA damage responses. AB - The Rad1 gene is evolutionarily conserved from yeast to human. The fission yeast Schizosaccharomyces pombe Rad1 ortholog promotes cell survival against DNA damage and is required for G(2)/M checkpoint activation. In this study, mouse embryonic stem (ES) cells with a targeted deletion of Mrad1, the mouse ortholog of this gene, were created to evaluate its function in mammalian cells. Mrad1 (-/-) ES cells were highly sensitive to ultraviolet-light (UV light), hydroxyurea (HU) and gamma rays, and were defective in G(2)/M as well as S/M checkpoints. These data indicate that Mrad1 is required for repairing DNA lesions induced by UV-light, HU and gamma rays, and for mediating G(2)/M and S/M checkpoint controls. We further demonstrated that Mrad1 plays an important role in homologous recombination repair (HRR) in ES cells, but a minor HRR role in differentiated mouse cells. PMID- 21637963 TI - Post-operative radiation improves survival in children younger than 3 years with intracranial ependymoma. AB - Concerns regarding long-term toxicities have led to the avoidance of post operative radiation (PORT) in young children with intracranial ependymoma. We investigated the association between post-operative radiation therapy and overall survival (OS) in children younger than 3 years and compared their survival to other age groups. The study sample from the SEER database included 804 patients with intracranial ependymoma, grades 2-3, and diagnosed between 1988 and 2005. OS was estimated using the Kaplan-Meier method, and hazard ratios (HR) and 95% confidence limits (CL) were calculated based on multivariable Cox proportional hazards models. A total of 804 patients were selected and PORT was administered to 35% of patients younger than 3 years. With a median follow-up of 3 years (range 0.1-18 years), the 3 year OS was 61% for children younger than 3 years, 83% for those ages 3-20 years, and 69% for patients older than 20 years (P < 0.001). In multivariable analysis, OS was significantly improved for patients receiving PORT (HR 0.8, 95% CL 0.6-0.9), and gross total resection (HR 0.6, 95% CL 0.5-0.8). Among children younger than 3 years, the 3 year OS was significantly greater among those who received PORT compared to those who did not (81% vs. 56%, respectively, P = 0.005). The majority of children younger than 3 years with intracranial ependymoma did not receive PORT. Children younger than 3 years who did not receive PORT had a relatively poor outcome, while those who received radiation therapy had a survival similar to older patients. PMID- 21637964 TI - Herpes simplex encephalitis in patients with cancer. AB - Case reports and animal models suggest that chemotherapy, corticosteroids and radiotherapy (RT) may increase the risk of herpes simplex encephalitis (HSE). We retrospectively examined cases of HSE at an academic hospital devoted to cancer care. Patients were identified by positive herpes simplex virus (HSV) polymerase chain reaction (PCR) in cerebrospinal fluid (CSF) or by brain pathology. There were seven patients with HSE over a 12 year period, four of whom had received cranial RT. During this time, a total of 997 patients were treated with cranial RT, suggesting a greater incidence than the expected risk of two to four cases per million people per year in the general population. Five patients had recently received chemotherapy and three were on dexamethasone. MRI findings were typical; four patients had bilateral anterior temporal lesions and three had unilateral temporal lesions. Four patients had a normal CSF white blood cell count, three of whom had prior RT and dexamethasone. Four patients were positive for HSV-1, and two for HSV-2. One patient had a negative CSF PCR for HSV, but autopsy confirmed active HSE. Though still rare, the risk of HSE may be increased in patients with cancer, especially in those receiving cranial RT. MRI findings were typical, but CSF white blood cell count was normal in four patients and one had negative CSF testing, suggesting that CSF results may be misleading in this population. PMID- 21637965 TI - A Case of Primary Myxoid Malignant Fibrous Histiocytoma of the Pancreas: Clinical, Histopathology, and Immunohistochemistry Findings. PMID- 21637966 TI - Expression of cell cycle regulators p21 and p27 as predictors of disease outcome in colorectal carcinoma. AB - BACKGROUND: Recent studies suggest that aberrations in cell cycle checkpoint controllers are a common feature in human malignancies and predict prognosis independent of stage. OBJECTIVES: This study correlated two cell cycle regulators (p27 and p21) with clinical and pathological variables in colorectal cancer (CRC) patients to assess their role as prognostic factors. PATIENTS AND METHODS: A series of 65 CRC patients were analyzed for p27 and p21 expression in their tumors using immunohistochemistry. RESULTS: Forty-six percent of tumors showed positive nuclear p27 expression, whereas 72% of cases were completely p21 negative. There were no significant correlations between p27 and p21 expression and gender, age, lymph node involvement, stage, and grade. However, p27 (but not p21) expression revealed highly significant correlation with tumor location (p < 0.01), depth of invasion (p < 0.03), and lympho-vascular invasion (p < 0.02). Tumors with high p27 expression showed a higher recurrence rate than tumors with no expression (p < 0.03). In Kaplan-Meier survival analysis, there was a significant (p = 0.046) difference in disease-free survival (DFS) between p27 positive and p27-negative tumors in favor of the latter. p21 did not show any predictive value of DFS (p < 0.7). Neither p27 nor p21 did predict disease specific survival (DSS) in Kaplan-Meier analysis, but DSS time was much shorter for p27-positive tumors. In multivariate (Cox) model, p27 lost its value as independent predictor of DFS, and none of the covariates were independent predictors of DSS. CONCLUSION: p27 expression seems to be more powerful than p21 expression in providing useful prognostic information in CRC, particularly in predicting the patients at high risk for recurrent disease. Larger cohort and longer follow-up are needed to fully elucidate the value of p27 (and p21) as independent predictors of disease outcome. PMID- 21637967 TI - A dual role for MYB60 in stomatal regulation and root growth of Arabidopsis thaliana under drought stress. AB - In response to environmental challenges, plant cells activate several signaling pathways that trigger the expression of transcription factors. Arabidopsis MYB60 was reported to be involved in stomatal regulation under drought conditions. Here, two splice variants of the MYB60 gene are shown to play a crucial role in stomatal movement. This role was demonstrated by over-expressing each variant, resulting in enhanced sensitivity to water deficit stress. The MYB60 splice variants, despite the fact that one of which lacks the first two exons encoding the first MYB DNA binding domain, both localize to the nucleus and promote guard cell deflation in response to water deficit. Moreover, MYB60 expression is increased in response to a low level of ABA and decreased in response to high level of ABA. At initial stage of drought stress, the plant system may modulate the root growth behavior by regulating MYB60 expression, thus promotes root growth for increased water uptake. In contrast, severe drought stress inhibits the expression of the MYB60 gene, resulting in stomatal closure and root growth inhibition. Taken together, these data indicate that MYB60 plays a dual role in abiotic stress responses in Arabidopsis through its involvement in stomatal regulation and root growth. PMID- 21637968 TI - Mesenchymal stem cells for cardiovascular regeneration. AB - Despite recent studies suggesting that the heart has instrinsic mechanisms of self-regeneration following myocardial infarction, it cannot regenerate itself to an optimal level. Mesenchymal stem cells (MSCs) are currently being investigated for regeneration of mesenchyme-derived tissues, such as bone, cartilage and tendon. In vitro evidence suggests that MSCs can also differentiate into cardiomyogenic and vasculogenic lineages, offering another cell source for cardiovascular regeneration. In vivo, MSCs may contribute to the re-growth and protection of vasculature and cardiomyocytes, mediated by paracrine actions, and/or persist within the myocardium in a differentiated state; although proof of cardiomyocytic phenotype and functional integration remains elusive. Herein, we review the evidence of MSCs as a cell source for cardiovascular regeneration, as well as their limitations that may prevent them from being effectively used in the clinic. PMID- 21637969 TI - Design and rationale of Japanese evaluation between Formula of Azelnidipine and amlodipine add on olmesartan to Get antialbuminuric effect study (J-FLAG) : evaluation of the antialbuminuric effects between calcium channel blocker with sympatholytic action in hypertensive patients with diabetes and albuminuria. AB - PURPOSE: Calcium channel blockers (CCBs) are recommended second-line antihypertensives for renin-angiotensin system (RAS) inhibitor-treated patients with chronic kidney disease (CKD), but they do not always ameliorate the progression of CKD. However, small clinical studies suggest that sympatholytic CCBs may protect against kidney injury. Therefore, a clinical trial was designed to test whether the sympatholytic CCB azelnidipine decreases the urinary albumin levels of CKD patients treated with the angiotensin receptor blocker olmesartan more potently than the widely-used non-sympatholytic CCB amlodipine. METHODS: A multi-center, open-labeled, randomized clinical intervention trial was designed to compare the antialbuminuric effect of azelnidipine (8-16 mg/day) and amlodipine (2.5-5 mg/day) in olmesartan-treated hypertensive (blood pressure 130 180/80-110 mmHg) patients with type 2 diabetes (fasting blood sugar >=126 mg/dL or treatment with antidiabetic agents) and albuminuria (urinary albumin/creatinine ratio >=30 mg/g). The primary study endpoint is the change in the urinary albumin/creatinine ratio after 12 months of treatment. CONCLUSIONS: The present trial is expected to clarify whether the sympatholytic CCB azelnidipine is a beneficial second-line choice for RAS inhibitor-treated hypertensive patients with CKD, such as diabetic nephropathy. PMID- 21637970 TI - Removal of genotoxicity in chlorinated secondary effluent of a domestic wastewater treatment plant during dechlorination. AB - PURPOSE: Dechlorination with tetravalent sulfur is widely used in wastewater treatment processes after chlorination. Dechlorination can remove certain genotoxic disinfection by-products (DBPs). However, the reactions occurring during dechlorination of chlorinated secondary effluent and their genotoxic chemicals are still very complex, and the related genotoxicity changes remain unknown. Therefore, the effects of dechlorination on genotoxicity in secondary effluent and its fractions and typical genotoxic chemical after chlorination were evaluated. METHODS: The dissolved organic matter in the secondary effluent sample was separated into four fractions with XAD-8 resin. Genotoxicity of secondary effluent and its fractions was evaluated by SOS/umu test, an ISO standard method. The concentration of typical genotoxic chemical named ofloxacin was determined by liquid chromatography with a mass spectrometer and a fluorescence detector. RESULTS: Dechlorination with the addition of Na(2)SO(3) notably decreased the genotoxicity in the chlorinated secondary effluent, especially in the presence of high ammonia nitrogen concentration in the sample before chlorination. The Na(2)SO(3) addition significantly decreased the genotoxicity of the secondary effluent and its genotoxic ofloxacin prior to chlorination. The genotoxicity in the fractions containing hydrophobic acids (HOA) increased after chlorination, while addition of Na(2)SO(3) decreased the genotoxicity induced by chlorination. Tryptophan found in HOA exhibited genotoxicity after chlorination, while dechlorination decreased the genotoxicity in chlorinated tryptophan induced by DBPs. CONCLUSIONS: Dechlorination was found to decrease the genotoxicity of chlorinated secondary effluent. The decrease was associated with the reduction of genotoxicity in genotoxic chemicals in secondary effluent prior to chlorination and DBPs. PMID- 21637971 TI - Salicylic acid minimizes nickel and/or salinity-induced toxicity in Indian mustard (Brassica juncea) through an improved antioxidant system. AB - INTRODUCTION: This study was hypothesized that salicylic acid elevates the level of antioxidant system that will protect plants from the stress generated by nickel and/or salinity. MATERIALS AND METHODS: Seeds of Brassica juncea were sown in sand amended with NiCl(2) (100 mg kg(-1)) and/or 15-day-old seedlings supplied for 3 days with NaCl (150 mM) and were then, at 20-day stage, sprayed with salicylic acid (10(-5) M) to assess selected morphological, physiological, and biochemical parameters at 30-day stage. RESULTS: The combination of Ni and NaCl proved most deleterious and exhibited significant decline in growth, leaf water potential, the level of pigments, and photosynthetic attributes. However, the follow-up treatment with salicylic acid detoxified the stress-generated damages caused by the combination (NiCl(2) and NaCl) and also significantly improved values for the above parameters. The NiCl(2) and/or NaCl increased electrolyte leakage, lipid peroxidation, and H(2)O(2) content but decreased the membrane stability index and activity of nitrate reductase and carbonic anhydrase. However, the salicylic acid treatment in the presence or absence of the stress improved the activity of nitrate reductase and carbonic anhydrase. The activity of antioxidative enzymes and the level of proline exhibited a significant increase in response to NiCl(2) and/or NaCl stress and which enhanced further with the spray of salicylic acid. CONCLUSIONS: It is concluded that the elevated level of antioxidative enzymes and level of proline might be responsible for minimizing the Ni and/or salinity-induced toxicity in Indian mustard which is manifested in terms of improved growth and photosynthesis. PMID- 21637972 TI - 2011--year of the bold step? PMID- 21637973 TI - Patients' understanding of clinical trials needs to be assessed in the context of understanding of overall goals of care. PMID- 21637974 TI - Improving the availability and accessibility of opioids for the treatment of pain: the International Pain Policy Fellowship. AB - Opioid analgesics are simultaneously indispensable medicines for the treatment of moderate to severe pain and are harmful when abused. The challenge for governments is to balance the obligation to prevent diversion, trafficking, and abuse of opioids with the equally important obligation to ensure their availability and accessibility for the relief of pain and suffering. Over the last 30 years, significant progress has been made toward improving access to opioids as measured by increasing global medical opioid consumption. However, this progress is marked by ongoing large disparities among countries, with most increases in medical opioid consumption attributed to high-income countries, not low- and middle-income countries (LMICs). The International Pain Policy Fellowship (IPPF) was developed by the Pain & Policy Studies Group, with the central goal of developing national leaders from LMICs and empowering them to improve availability and accessibility of opioids for the treatment of pain. To date, two classes of fellows have been selected, representing 17 fellows from 15 countries. Progress achieved by the leadership of three fellows from Sierra Leone, Colombia, and Serbia is highlighted in this paper. The fellows from each country were successful at initiating collaboration with relevant governmental bodies, national authorities, and professional societies, which resulted in a new supply of oral opioids in Sierra Leone and Serbia, and improvements in the distribution of already available opioids in Colombia. All fellows were instrumental in facilitating evaluation of national policy. The IPPF program empowers fellows with the necessary knowledge, skills, and guidance to improve the availability and accessibility of opioids for the treatment of pain. PMID- 21637975 TI - Utilization of hypnotic medication in the context of cancer: predictors and frequency of use. AB - OBJECTIVE: The present study aims to document the frequency of use of hypnotic medication among a large sample of randomly selected patients having been treated for various types of cancer, as well as to identify the sociodemographic, psychosocial, and medical factors that characterize the users of this type of medication. METHODS: Five thousand patients who had received treatment for breast, prostate, lung, or colorectal cancer at the L'Hotel-Dieu de Quebec were solicited by mail to take part in this study. Among these patients, 1,984 (39.7%) agreed to complete a battery of questionnaires. RESULTS: Overall, 22.6% of the patients were currently consuming hypnotic medication. Factors associated with a greater utilization of hypnotic medication were older age, greater difficulties initiating sleep, more stressful life events experienced in the past 6 months, higher levels of anxiety, past or current psychological difficulties, poorer role functioning, less severe urinary symptoms, greater use of opioids, and past or current chemotherapy treatments. CONCLUSIONS: These results are consistent with those of previous studies conducted in cancer patients in showing high rates of hypnotic medication use. Moreover, this study identified several factors that might help identify persons at risk of using this type of medication and, therefore, to experience the potential negative effects of chronic hypnotics use. PMID- 21637976 TI - Isolation and characterization of a new cellulosome-producing Clostridium thermocellum strain. AB - The anaerobic thermophilic bacterium, Clostridium thermocellum, is a potent cellulolytic microorganism that produces large extracellular multienzyme complexes called cellulosomes. To isolate C. thermocellum organisms that possess effective cellulose-degrading ability, new thermophilic cellulolytic strains were screened from more than 800 samples obtained mainly from agriculture residues in Thailand using microcrystalline cellulose as a carbon source. A new strain, C. thermocellum S14, having high cellulose-degrading ability was isolated from bagasse paper sludge. Cellulosomes prepared from S14 demonstrated faster degradation of microcrystalline cellulose, and 3.4- and 5.6-fold greater Avicelase activity than those from C. thermocellum ATCC27405 and JW20 (ATCC31449), respectively. Scanning electron microscopic analysis showed that S14 had unique cell surface features with few protuberances in contrast to the type strains. In addition, the cellulosome of S14 was resistant to inhibition by cellobiose that is a major end product of cellulose hydrolysis. Saccharification tests conducted using rice straw soaked with sodium hydroxide indicated the cellulosome of S14 released approximately 1.5-fold more total sugars compared to that of ATCC27405. This newly isolated S14 strain has the potential as an enzyme resource for effective lignocellulose degradation. PMID- 21637977 TI - Anatomic and functional correlation of frequency-doubling technology perimetry (FDTP) in multiple sclerosis. AB - To study the correlation between the results of frequency-doubling technology perimetry (FDTP), visual function (visual acuity, contrast vision, standard automated perimetry (SAP)) and the thickness of the retinal nerve fiber layer (RNFL) throughout the course of multiple sclerosis (MS). Sixty-six eyes of thirty three patients suffering from MS were chosen. Thirty-five eyes had a previous history of optic neuritis (ON group) and thirty-one eyes had no previous history of optic neuritis (non-ON group). The FDTP was performed with the N-30 screening program. Visual acuity was determined with the Snellen scale and the ETDRS (Early Treatment Diabetic Retinopathy Study) scale, the contrast vision with the Pelli Robson and Sloan tests and the SAP with the Humphrey 750 perimeter. The thickness of the RNFL was measured using the STRATUS OCTTM optical coherence tomography (OCT). The visual field FDTP was divided into three sectors corresponding to the three SAP sectors and to the three RNFL quadrants of the OCT. The FDTP was significantly correlated (P<0.0001) to the contrast vision and to the SAP results (mean deviation (MD) and the different sectors among themselves). In the ON group, the MD FDTP was significantly correlated to the average RNFL thickness (r=0.44, P=0.0091). A decrease of 5 decibels (dB) of the MD FDTP corresponded to a decrease of 11.7 MUm of the average RNFL thickness (Y=2.34*X+87.5). The strong correlation with SAP and RNFL confirms the value of FDTP in assessing optic nerve damage throughout the course of MS. PMID- 21637978 TI - Photodynamic therapy for unilateral idiopathic peripapillary choroidal neovascularization in a child. AB - A 10-year-old girl presented with visual loss in her right eye. Best-corrected visual acuity was 20/50 in the right eye and 20/20 in the left eye. Dilated funduscopic examination revealed a yellowish elevated lesion near the optic disc with macular edema and hemorrhage on the inferotemporal vascular arcade in the right eye. Fluorescein angiography showed a hyperfluorescent lesion consistent with choroidal neovascularization (CNV) and optical coherence tomography (OCT) showed a peripapillary lesion with subretinal fluid elevating the neurosensory retina in the macular area. With a diagnosis of unilateral idiopathic peripapillary CNV, the patient underwent photodynamic therapy (PDT) with verteporfin. At the four month follow-up, visual acuity increased to 20/25 and OCT showed peripapillary scar formation and total resolution of the subretinal fluid. At the one year follow-up, visual acuity and fundus were stable without recurrence of the lesion. PDT for peripapillary CNV in children has not previously been reported. Although it seems to be a favorable treatment option for peripapillary lesions, there is a lack of knowledge about long-term follow-up in pediatric cases. PMID- 21637979 TI - Epicardial adipose tissue thickness predicts descending thoracic aorta atherosclerosis shown by multidetector computed tomography. AB - Human epicardial adipose tissue (EAT) is a type of visceral adipose tissue functioning as an endocrine organ by secreting hormones and adipocytokines which have an important role in the atherosclerotic process. In this study, we aimed to assess the relationship between EAT measured by dual source multidetector computed tomography (MDCT) and descending thoracic aorta (DTA) atherosclerosis. A total of 148 patients who underwent MDCT for the evaluation of coronary artery disease were enrolled in this study. Thickness of the EAT was measured on contrast enhanced multiplanar reformat images with parasternal short axis view at basal, mid-ventricular and apical levels and horizontal long axis view. The atherosclerotic plaque was scored from 0 to 4 points by the percentage of the luminal surface at the cross sectional area of proximal, mid and distal segments of descending aorta. Among the study population, 84 (56.8%) were male and age was (mean +/- standart deviation) 56.9 +/- 11.7 years. In patients with critical coronary atherosclerosis, DTA atherosclerosis had a significant relationship with EAT (P = 0.012). Multivariate linear regression analysis revealed that in addition to critical coronary stenosis, age and total epicardial fat thickness were associated with aortic atherosclerosis (beta value, 0.058 and 0.035; t value, 4.74 and 2.28, respectively; P < 0.05) after adjustment for traditional cardiovascular risk factors. In this study we demonstrated that atherosclerotic plaque burden of DTA was associated with the amount of EAT thickness among patients with suspected CAD shown by MDCT. Further large scale prospective studies are needed to address the interaction of EAT as well as the mediators of inflammation and adipocytokines with the development of atherosclerotic plaques in aorta and effects on cardiovascular outcomes. PMID- 21637980 TI - Optimization of energy level for coronary angiography with dual-energy and dual source computed tomography. AB - Dual-energy computed tomography (DE-CT) uses polyenergetic X-rays at 100- and 140 kVp tube energy, and generates 120-kVp composite images that are referred to as polyenergetic images (PEIs). Moreover, DE-CT can produce monoenergetic images (MEIs) at any effective energy level. We evaluated whether the image quality of coronary angiography is improved by optimizing the energy levels of DE-CT. We retrospectively evaluated data sets obtained from 24 consecutive patients using cardiac DE-CT at 100- and 140-kVp tube energy with a dual-source scanner. Signal to-noise ratios (SNRs) were evaluated in the left ascending coronary artery in PEIs, and in MEIs reconstructed at 40, 50, 60, 70, 80, 90, 100, 130, 160 and 190 keV. Energy levels of 100, 120 and 140 kVp generated the highest SNRs in PEIs from 10, 12 and 2 patients, respectively, at 60, 70 and 80 keV in MEIs from 2, 10 and 10 patients, respectively, and at 90 and 100 keV in those from one patient each. Optimization of the energy level for each patient increased the SNR by 16.6% in PEIs (P < 0.0001) and by 18.2% in MEIs (P < 0.05), compared with 120-kVp composite images. The image quality of coronary angiography using DE-CT can be improved by optimizing the energy level for individual patients. PMID- 21637981 TI - Automatic centerline extraction of coronary arteries in coronary computed tomographic angiography. AB - Coronary computed tomographic angiography (CCTA) is a non-invasive imaging modality for the visualization of the heart and coronary arteries. To fully exploit the potential of the CCTA datasets and apply it in clinical practice, an automated coronary artery extraction approach is needed. The purpose of this paper is to present and validate a fully automatic centerline extraction algorithm for coronary arteries in CCTA images. The algorithm is based on an improved version of Frangi's vesselness filter which removes unwanted step-edge responses at the boundaries of the cardiac chambers. Building upon this new vesselness filter, the coronary artery extraction pipeline extracts the centerlines of main branches as well as side-branches automatically. This algorithm was first evaluated with a standardized evaluation framework named Rotterdam Coronary Artery Algorithm Evaluation Framework used in the MICCAI Coronary Artery Tracking challenge 2008 (CAT08). It includes 128 reference centerlines which were manually delineated. The average overlap and accuracy measures of our method were 93.7% and 0.30 mm, respectively, which ranked at the 1st and 3rd place compared to five other automatic methods presented in the CAT08. Secondly, in 50 clinical datasets, a total of 100 reference centerlines were generated from lumen contours in the transversal planes which were manually corrected by an expert from the cardiology department. In this evaluation, the average overlap and accuracy were 96.1% and 0.33 mm, respectively. The entire processing time for one dataset is less than 2 min on a standard desktop computer. In conclusion, our newly developed automatic approach can extract coronary arteries in CCTA images with excellent performances in extraction ability and accuracy. PMID- 21637982 TI - Right ventricular function declines after cardiac surgery in adult patients with congenital heart disease. AB - Right ventricular function (RVF) is often selectively declined after coronary artery bypass graft surgery. In adult patients with congenital heart disease (CHD) the incidence and persistence of declined RVF after cardiac surgery is unknown. The current study aimed to describe RVF after cardiac surgery in these patients. Adult CHD patients operated between January 2008 and December 2009 in the Academic Medical Centre in Amsterdam were studied. Clinical characteristics, laboratory tests, surgical data and intensive care unit outcome were obtained from medical records. RVF was measured by trans-thoracic echocardiography (TTE) and expressed by tricuspid annular plane systolic excursion (TAPSE), tissue Doppler imaging (RV S') and myocardial performance index (MPI) pre-operatively and direct, at intermediate and late follow up. Of a total of 185 operated, 86 patients (mean age 39 +/- 13 years, 54% male) had echo data available. There was a significant fall in RVF after cardiac surgery. TAPSE and RV S' were significantly higher and MPI was significantly lower pre-operatively compared to direct post-operative values (TAPSE 22 +/- 5 versus 13 +/- 3 mm (P < 0.01), RV S' 11 +/- 4 versus 8 +/- 2 cm/s (P < 0.01) and MPI 0.36 +/- 0.14 vs 0.62 +/- 0.25; P < 0.01). There were no significant differences in left ventricular function pre operatively compared to post-operative values. Right-sided surgery was performed in 33, left-sided surgery in 37 and both sided surgery in 16 patients. Decline in RVF was equal for those groups. Patients with severe decline in RVF, were patients who underwent tricuspid valve surgery. Decline in RVF was associated with post-operative myocardial creatine kinase level and maximal troponin T level. There was no association between decline in RVF and clinical outcome on the intensive care unit. 18 months post-operatively, most RVF parameters had recovered to pre-operative values, but TAPSE which remained still lower (P < 0.01). CHD patients have a decline in RVF directly after cardiac surgery, regardless the side of surgery. Although a gradual improvement was observed, complete recovery was not seen 18 months post-operatively. PMID- 21637983 TI - Intratumoural hydrogen peroxide injection during resection of metastatic vertebral melanoma. Technical note. AB - It is well recognized that the surgical removal of vertebral metastases of several malignant tumours often causes extensive bleeding, increasing the difficulties for surgeons and the risks for patients. Among the different subtypes of malignancies involved, melanoma has an extensive vascularity, and its resection is frequently challenging because of a continuous and annoying bleeding. A 55-year-old patient with a history of treated melanoma was operated on urgently for a severe T10-11 spinal cord compression by a huge metastasis involving the peridural space and the vertebral bone. During the resection, we injected, in successive stages, a total of 5 ml of 3% hydrogen peroxide (H(2)O(2)) solution, with good control of tumour bleeding. The patient had no problems related to the procedure, and 6 months after the surgery he is able to perform sustained walking. The aim of this manuscript is to report a safe, simple, effective, inexpensive and readily available technique for intraoperative haemostasis that could allow the control of tumour bleeding and facilitate its excision. PMID- 21637984 TI - Saliva in perimenopausal and early postmenopausal women. A 2-year follow-up study. AB - This study aims to investigate salivary flow and biochemical constituents of menopausal-age women with the hypothesis that women using hormone therapy (HT) might present better saliva values than non-users. Two hundred HT users and 200 non-users were selected at random from a cohort study of 3,173 peri- and postmenopausal women and invited to a 2-year clinical follow-up study. Clinical examination with saliva sampling was made at baseline and 2 years later. Salivary total protein, albumin, and immunoglobulin (Ig) concentrations were analyzed. Final material included 106 consistent HT users and 55 non-users. Backward logistic regression analysis was made to determine the risk factors for higher or lower than medium salivary protein values. No difference was seen in salivary flow rate, total protein, and IgA values between baseline and follow-up measurements or between the groups. Albumin, IgG, and IgM concentrations were significantly lower in the 2-year samples of the HT group when compared with baseline. IgA and IgM values were higher in the non-HT 2-year samples when compared with the corresponding HT samples. The only significant explanatory factor for higher than median salivary albumin concentration was the number of teeth both at baseline and 2 years later. HT possibly improved epithelial integrity since the concentrations of serum components albumin, IgG, and IgM decreased during the follow-up. HT as such does not seem to affect saliva, although it may modify it. The clinical relevance of these results needs to be assessed in future studies. PMID- 21637985 TI - Cancer causes & control. PMID- 21637986 TI - Menopausal hormone therapy and risk of ovarian cancer in the European prospective investigation into cancer and nutrition. AB - The association between menopausal hormone therapy (HT) and risk of ovarian cancer was assessed among 126,920 post-menopausal women recruited into the European Prospective Investigation into Cancer and Nutrition. After an average of 9-year follow-up, 424 incident ovarian cancers were diagnosed. Cox models adjusted for body mass index, smoking status, unilateral ovariectomy, simple hysterectomy, age at menarche, number of full-term pregnancies, and duration of oral contraceptives were used. Compared with baseline never use, current use of any HT was positively associated with risk (HR [hazard ratio], 1.29; 95% CI [confidence interval], 1.01-1.65), while former use was not (HR, 0.96; 95% CI, 0.70-1.30). Current estrogen-only HT was associated with a 63% higher risk (HR, 1.63; 95% CI, 1.08-2.47), while current estrogen plus progestin was associated with a smaller and non-significant higher risk (HR, 1.20; 95% CI, 0.89-1.62). Use of tibolone was associated with a twofold greater risk (HR, 2.19; 95% CI, 1.06 4.50), but was based on small numbers. In conclusion, women who currently use HT have a moderate increased risk of ovarian cancer, and which may be stronger for estrogen-only than estrogen plus progestin preparations. PMID- 21637987 TI - Hat, shade, long sleeves, or sunscreen? Rethinking US sun protection messages based on their relative effectiveness. AB - BACKGROUND: Sun protection messages in the United States emphasize sunscreen use, although its efficacy in skin cancer prevention remains controversial. METHODS: We used data from NHANES 2003-2006, restricted to adult whites (n = 3,052) to evaluate how Americans protect themselves from the sun. Participants completed questionnaires on the frequency with which they used sunscreen, wore a hat, long sleeves, or stayed in the shade, in addition to the number of sunburns in the past year. RESULTS: Although using sunscreen is the most common sun protective behavior (30%), frequent sunscreen use was not associated with fewer sunburns. However, the odds of multiple sunburns were significantly lower in individuals who frequently avoided the sun by seeking shade (OR = 0.70, p < 0.001) or wearing long sleeves (OR = 0.73, p = 0.01). CONCLUSIONS: Our findings suggest that shade and protective clothing may be more effective than sunscreen, as typically used by Americans. PMID- 21637988 TI - Risk factors for chronic kidney diseases may include periodontal diseases, as estimated by the correlations of plasma pentraxin-3 levels: a case-control study. AB - BACKGROUND: Pentraxins are classical mediators of inflammation and markers of acute-phase reactions. Pentraxin-3 (PTX3) is believed to be a true independent indicator of disease activity. It has been associated with clinical outcomes in incident chronic kidney disease (CKD) and periodontal diseases. Periodontitis is lately being considered as a risk factor for CKD. However, no data are available on elevated PTX3 in patients with CKD associated with periodontitis. METHOD: Sixty subjects were divided into three groups (n = 20) based on glomerular filtration rate (GFR) and periodontal parameters: healthy (group-1), CKD (group 2), and CKD with periodontitis (group-3). Plasma samples obtained from each patient were quantified for PTX3 using Enzyme-linked Immunosorbent Assay (ELISA). RESULTS: Both patient groups with CKD had higher plasma PTX3 concentrations than control subjects. However, there was no significant difference between the two groups (groups 2 and 3). In all groups, plasma PTX3 correlated positively with periodontal parameters. Group 3 patients had higher concentrations of PTX3 (6.338 ng/ml) than group 2 (5.41 ng/ml) and group 1 (1.835 ng/ml). CONCLUSIONS: Within the limits of the present study, the difference in plasma PTX3 levels between groups 2 and 3 was not found to be statistically significant (P > 0.05). However, as PTX3 values correlated positively with periodontal parameters, this model could contribute to identifying individuals with periodontitis at high risk of CKD. Thus, periodontal disease could serve as a risk factor for developing CKD. Further large-scale studies nullifying the confounders for CKD are warranted to confirm positive results. PMID- 21637989 TI - Fatal catheter-related bacteremia due to Alcaligenes (Achromobacter) xylosoxidans in a hemodialysis patient. PMID- 21637990 TI - Effects of temporarily disrupting BBB on activity-induced manganese-dependent functional MRI. AB - This study further investigates the influence of temporarily disrupting the blood brain barrier (BBB) on the level of manganese used in AIM fMRI other than the recognized function of allowing that substance to enter into the activated brain regions more effectively during the BBB opening. We injected manganese into Wistar rats through ICA following the disruption of BBB with mannitol in a functional MRI test of the visual cortex. Through comparing MRI signal intensity and manganese contents in the visual cortex of rats received visual stimuli of unequal degree after the restoration of BBB, we found that the signal in the visual cortex could be further enhanced on T1WI given visual stimulation after the restoration of BBB. Temporary BBB disruption has an additional advantage in allowing Mn(2+) to enter the CSF or brain for later transference to the activated brain area. So the dosage of manganese in AIM fMRI could be minimized by extending the stimulus. PMID- 21637991 TI - Combined chlorhexidine and PVP-I decontamination of human donor eyes prior to corneal preservation. AB - The aim of this study was to report the efficacy of adding chlorhexidine to the protocol for decontamination of human donor globes prior to excision of corneo scleral rims for future keratoplasty procedures. In 2005, chlorhexidine was introduced by our eye bank as an additional step in the protocol for decontaminating human donor globes. After 5 years, we prospectively evaluated the number of contaminations. Out of 2,891 globes included in our study, 2,663 globes were processed, of which 36 (1.4%) were considered contaminated. Seventeen contaminations (0.6%) were detected by culturing limbal swabs, directly after decontamination, eight (0.3%) by visible discoloration of the culture medium carrying a corneo-scleral rim, and eleven (0.4%) after inoculation of the culture medium on blood agar plates. Importantly, after 4 weeks of incubation, none of the aerobic and anaerobic cultures taken from the secondary 'transport medium' (dextran containing medium used to transport corneal tissue to the transplantation centre) showed microbiological growth. In conclusion, the combined use of 0.02% chlorhexidine and 0.5% povidone-iodine may allow decontamination of donor globes to a level at which the risk of tissue contamination at the time of transplantation is minimized, while corneal viability is preserved. PMID- 21637993 TI - The role of working equines to livelihoods in current day campesino hill-slope communities in central Mexico. AB - Small-holder campesino agriculture is based on the diversified use of resources and off-farm work. Working equines have a multifunctional character and sustain the diversification of livelihoods having different values as assets or providing services. The objective was to identify the role of working equines in current diversification strategies in the livelihoods of campesino families in a hill slope community in central Mexico within livelihoods analysis. Thirty-one variables related to ownership and use of working equines were analysed by cluster analysis and descriptive statistics contrasting the presence of equines in the diversification of livelihoods. Four groups were identified, determined mainly by age of farmer and number of family members who utilise equines. Results show these systems diversify in response to conditions of risk or to take advantage of opportunities, such that a balance is reached by resorting to off farm activities without the total loss of components of the farming system. Two main situations were found in relation to working equines: the disappearance and change of functions of the large equines (mules), and the adaptation of small equines (donkeys) to the new conditions. It is concluded that there is a process of adaptation in hill-slope campesino farms such that large equines are less present in farms that have moved towards more diversification, but are kept in those farms less diversified. The use of equines for draught force in agricultural production and as pack animals continues, as is the presence of small livestock (sheep and poultry) irrespective of the context of the farm. PMID- 21637994 TI - Factors influencing the prevalence of subclinical mastitis in lactating dromedary camels in Riyadh Region, Saudi Arabia. AB - The purpose of this study was to determine the prevalence of subclinical mastitis in camels in Riyadh, Saudi Arabia and the factors influencing its incidence. A total of 740 quarter milk samples were collected from 47 camel herds belonging to Majahim, Maghatir, Shu'l, and Sufer breeds. California mastitis test (CMT) was used as a screening test for subclinical mastitis. Samples giving negative or trace CMT scores (0) were assigned to healthy quarters, while those giving positive scores of 1+ to 3+ were assigned to subclinically affected quarters. Logistic regression was used to assess the association of breed, parity, and stage of lactation with the prevalence of subclinical mastitis. Milk fat, protein, lactose, solid nonfat percentages and Na, Ca, and K concentrations were compared in CMT-positive versus healthy quarters. One third (33%) of tested quarters had subclinical mastitis based on CMT. The estimated probability of subclinical mastitis with the combined effects of breed, parity, and stage of lactation ranged from 15.8% to 54.6%. The risk of subclinical mastitis increased significantly with parity and with the early stage of lactation. The Shu'l breed had significantly higher prevalence of subclinical mastitis than other breeds. Significant decreases in protein, lactose, and solid nonfat, Ca and K concentrations and increase in Na concentrations were associated with subclinical mastitis. In conclusion, subclinical mastitis is prevalent in Saudi camels, and its incidence is influenced by breed, parity, and stage of lactation. PMID- 21637995 TI - Genetic diversity analysis of abiotic stress response gene TaSnRK2.7-A in common wheat. AB - Sucrose non-fermenting1-related protein kinase 2 (SnRK2) plays a key role in plant stress signaling transduction pathways. In this study, one copy of TaSnRK2.7, a SnRK2 member of common wheat, was isolated and characterized for nucleotide diversity among 45 wheat accessions with different stress-response features. Most of the accessions were elite wheat cultivars, which had been subject to population bottlenecks and intensive selection during breeding. Nucleotide and haplotype diversity across the entire TaSnRK2.7-A region was 0.00076 and 0.590, respectively, and diversity in non-coding regions was higher than that in coding regions. Sliding-window analysis showed variable levels of nucleotide variation along the entire TaSnRK2.7-A region; the sixth intron and ninth exon represented variation-enriched regions. As predicted, neutrality tests revealed that population bottlenecks or purifying selection had acted on the TaSnRK2.7-A gene, a relatively conserved gene. Furthermore, strong linkage disequilibrium between SNP loci extends across the entire TaSnRK2.7-A region. These findings demonstrate that the TaSnRK2.7-A genomic region has evolved under extensive selection pressure during crop breeding. PMID- 21637996 TI - VDR haploinsufficiency impacts body composition and skeletal acquisition in a gender-specific manner. AB - The vitamin D receptor (VDR) is crucial for virtually all of vitamin D's actions and is thought to be ubiquitously expressed. We hypothesized that disruption of one allele of the VDR gene would impact bone development and would have metabolic consequences. Body composition and bone mass (BMD) in VDR heterozygous (VDR HET) mice were compared to those obtained in male and female VDR KO and WT mice at 8 weeks of age. Male mice were also evaluated at 16 weeks, and bone marrow mesenchymal stem cell (MSC) differentiation was evaluated in VDR female mice. Additionally, female VDR HET and WT mice received intermittent PTH treatment or vehicle (VH) for 4 weeks. BMD was determined at baseline and after treatment. MRI was done in vivo at the end of treatment; MUCT and bone histomorphometry were performed after killing the animals. VDR HET male mice had normal skeletal development until 16 weeks of age but showed significantly less gain in fat mass than WT mice. In contrast, female VDR HET mice showed decreased total-body BMD at age 8 weeks but had a normal skeletal response to PTH. MSC differentiation was also impaired in VDR HET female mice. Thus, female VDR HET mice show early impairment in bone acquisition, while male VDR HET mice exhibit a lean phenotype. Our results indicate that the VDR HET mouse is a useful model for studying the metabolic and skeletal impact of decreased vitamin D sensitivity. PMID- 21637998 TI - Enhanced enantioselective recognition with diastereoisomeric BINOL based chiral fluorescent boronic acid sensors. AB - We prepared the diastereoisomers of BINOL based bisboronic acid chiral probes (the probes are with dual chirogenic centers) for enantioselective recognition of chiral analytes, such as tartaric acids, D-sorbitol, etc. We found the diastereoisomeric probes give different emission intensity-pH profiles, a phenomenon was reported, probably, for the first time. We found that with the second chirogenic center, the selectivity of the probes toward chiral analytes can be improved. For example, the diastereoisomeric probes give drastically different response to D-sorbitol, the same selectivity was not found for the BINOL bisboronic acid probes with single chirogenic center. Our result with the diastereoisomeric probes is helpful for design of new chiral molecular probes to enhance the selectivity of the boronic acid sensors toward chiral analytes. PMID- 21637997 TI - Adverse reactions and drug-drug interactions in the management of women with postmenopausal osteoporosis. AB - The pharmacological management of disease should involve consideration of the balance between the beneficial effects of treatment on outcome and the probability of adverse effects. The aim of this review is to explore the risk of adverse drug reactions and drug-drug interactions with treatments for postmenopausal osteoporosis. We reviewed evidence for adverse reactions from regulatory documents, randomized controlled trials, pharmacovigilance surveys, and case series. Bisphosphonates are associated with gastrointestinal effects, musculoskeletal pain, and acute-phase reactions, as well as, very rarely, atrial fibrillation, atypical fracture, delayed fracture healing, osteonecrosis of the jaw, hypersensitivity reactions, and renal impairment. Cutaneous effects and osteonecrosis of the jaw are of concern for denosumab (both very rare), though there are no pharmacovigilance data for this agent yet. The selective estrogen receptor modulators are associated with hot flushes, leg cramps, and, very rarely, venous thromboembolism and stroke. Strontium ranelate has been linked to hypersensitivity reactions and venous thromboembolism (both very rare) and teriparatide with headache, nausea, dizziness, and limb pain. The solidity of the evidence base depends on the frequency of the reaction, and causality is not always easy to establish for the very rare adverse reactions. Drug-drug interactions are rare. Osteoporosis treatments are generally safe and well tolerated, though they are associated with a few very rare serious adverse reactions. While these are a cause for concern, the risk should be weighed against the benefits of treatment itself, i.e., the prevention of osteoporotic fracture. PMID- 21637999 TI - Candidacy of a chitin-inducible gibberellin-responsive gene for a major locus affecting plant height in rice that is closely linked to Green Revolution gene sd1. AB - Appropriate plant height is crucial for lodging resistance to improve the rice crop yield. The application of semi-dwarf 1 led to the green revolution in the 1960s, by predominantly increasing the rice yield. However, the frequent use of single sd1 gene sources may cause genetic vulnerability to pests and diseases. Identifying useful novel semi-dwarf genes is important for the genetic manipulation of plant architecture in practical rice breeding. In this study, introgression lines derived from two parents contrasting in plant height, Zhenshan 97 and Pokkali were employed to locate a gene with a large effect on plant height by the bulk segregant analysis method. A major gene, ph1, was mapped to a region closely linked to sd1 on chromosome 1; the additive effects of ph1 were more than 50 cm on the plant height and 2 days on the heading date in a BC(4)F(2) population and its progeny. ph1 was then fine mapped to BAC AP003227. Gene annotation indicated that LOC_OS01g65990 encoding a chitin-inducible gibberellin-responsive protein (CIGR), which belongs to the GRAS family, might be the right candidate gene of ph1. Co-segregation analysis of the candidate gene derived marker finally confirmed its identity as the candidate gene. A higher expression level of the CIGR was detected in all the tested tissues in tall plants compared to those of short plants, especially in the young leaf sheath containing elongating tissues, which indicated its importance role in regulating plant height. ph1 showed a tremendous genetic effect on plant height, which is distinct from sd1 and could be a new resource for breeding semi-dwarf varieties. PMID- 21638000 TI - Genetic diversity and population structure in cultivated sunflower and a comparison to its wild progenitor, Helianthus annuus L. AB - Crop germplasm collections are valuable resources for ongoing plant breeding efforts. To fully utilize such collections, however, researchers need detailed information about the amount and distribution of genetic diversity present within collections. Here, we report the results of a population genetic analysis of the primary gene pool of sunflower (Helianthus annuus L.) based on a broad sampling of 433 cultivated accessions from North America and Europe, as well as a range wide collection of 24 wild sunflower populations. Gene diversity across the cultivars was 0.47, as compared with 0.70 in the wilds, indicating that cultivated sunflower harbors roughly two-thirds of the total genetic diversity present in wild sunflower. Population structure analyses revealed that wild sunflower can be subdivided into four genetically distinct population clusters throughout its North American range, whereas the cultivated sunflower gene pool could be split into two main clusters separating restorer lines from the balance of the gene pool. Use of a maximum likelihood method to estimate the contribution of the wild gene pool to the cultivated sunflower germplasm revealed that the bulk of the cultivar diversity is derived from two wild sunflower population genetic clusters that are primarily composed of individuals from the east-central United States, the same general region in which sunflower domestication is believed to have occurred. We also identified a nested subset of accessions that capture as much of the allelic diversity present within the sampled cultivated sunflower germplasm collection as possible. At the high end, a core set of 288 captured nearly 90% of the alleles present in the full set of 433, whereas a core set of just 12 accessions was sufficient to capture nearly 50% of the total allelic diversity present within this sample of cultivated sunflower. PMID- 21638001 TI - Fine-mapping of the woolly gene controlling multicellular trichome formation and embryonic development in tomato. AB - Trichomes are small hairs that originate from the epidermal cells of nearly all land plants, and they exist in unicellular and multicellular forms. The regulatory pathway of unicellular trichomes in Arabidopsis is well characterized. However, little is known about the multicellular trichome formation in tomato (Solanum lycopersicum). The woolly (Wo) gene controls multicellular trichome initiation and leads to embryonic lethality when homozygous in tomato. To clone and characterize Wo, the gene was fine-mapped to a DNA fragment of ~200 kb using the map-based cloning strategy. A series of sequence-based molecular markers, including simple sequence repeat, sequence characterized amplified region, and cleaved amplified polymorphic sequence were utilized in this study. Analysis of the sequence indicated that this region carries 19 putative open reading frames. These results will provide not only the important information for the isolation and characterization of Wo but also the starting point for studying the regulatory pathway responsible for trichome formation and embryonic lethality in tomato. PMID- 21638002 TI - Fractionation and speciation of arsenic in fresh and combusted coal wastes from Yangquan, northern China. AB - In this study, the content and speciation of arsenic in coal waste and gas condensates from coal waste fires were investigated, respectively, using the digestion and sequential extraction methods. The fresh and fired-coal waste samples were collected from Yangquan, which is one of the major coal production regions in northern China. High-performance liquid chromatography-inductively coupled plasma-mass spectrometry (HPLC-ICP-MS) was used to determine the concentrations of four major arsenic species [As(III), As(V), monomethylarsonic acid (MMA) and dimethylarsenic acid (DMA)] in the extracts, while ICP-MS was used to measure total As content. Arsenic content in the investigated coal wastes and the condensate ranges between 23.3 and 69.3 mg/kg, which are higher than its reported average content in soils. Arsenic in coal waste exists primarily in the residual fraction; this is followed in decreasing order by the organic matter bound, Fe-Mn oxides-bound, exchangeable, carbonates-bound, and water-soluble fractions. The high content of arsenic in the condensates indicates that combustion or spontaneous combustion is one of the major ways for arsenic release into the environment from coal waste. About 15% of the arsenic in the condensate sample is labile and can release into the environment under leaching processes. The water extractable arsenic (WEA) in the fresh coal waste, fired coal wastes, and the condensate varied between 14.6 and 341 MUg/kg, with As(V) as the major species. Furthermore, both MMA and DMA were found in fresh coal wastes, fired coal wastes, and the condensate. PMID- 21638003 TI - Functionally referential and intentional communication in the domestic dog: effects of spatial and social contexts. AB - In apes, four criteria are set to explore referential and intentional communication: (1) successive visual orienting between a partner and distant targets, (2) the presence of apparent attention-getting behaviours, (3) the requirement of an audience to exhibit the behaviours, and (4) the influence of the direction of attention of an observer on the behaviours. The present study aimed at identifying these criteria in behaviours used by dogs in communicative episodes with their owner when their toy is out of reach, i.e. gaze at a hidden target or at the owner, gaze alternation between a hidden target and the owner, vocalisations and contacts. In this study, an additional variable was analysed: the position of the dog in relation to the location of the target. Dogs witnessed the hiding of a favourite toy, in a place where they could not get access to. We analysed how dogs engaged in communicative deictic behaviours in the presence of their owner; four heights of the target were tested. To control for the motivational effects of the toy on the dogs' behaviour and for the referential nature of the behaviours, observations were staged where only the toy or only the owner was present, for one of the four heights. The results show that gazing at the container and gaze alternation were used as functionally referential and intentional communicative behaviours. Behavioural patterns of dog position, the new variable, fulfilled the operational criteria for functionally referential behaviour and a subset of operational criteria for intentional communication: the dogs used their own position as a local enhancement signal. Finally, our results suggest that the dogs gazed at their owner at optimal locations in the experimental area, with respect to the target height and their owner's (or their own) line of gaze. PMID- 21638004 TI - Geographic differences in the increasing ESRD rate have disappeared in Japan. AB - BACKGROUND: We previously showed that there are marked geographic differences in the incidence of end-stage renal disease (ESRD) within Japan. In addition, the use of renin-angiotensin system inhibitors was found to be inversely correlated with the increasing ESRD rate. It was recently demonstrated that the incidence of ESRD due to diabetic nephropathy is declining in both Europe and USA. Therefore, we investigated the increasing ESRD rate and its geographic difference in Japan. METHODS: Each year, the Japanese Society for Dialysis Therapy reports the numbers of patients initiating maintenance dialysis therapy in each prefecture of Japan. We used old (1984-1991) and recent (2001-2008) data to compare the increasing ESRD rate, which was estimated from the slope of the regression line of the annual incidence corrected for population, between the two periods in 11 regions of Japan. RESULTS: Increasing ESRD rate almost halved, from 11.1 +/- 5.6 to 5.4 +/- 0.7/million per year from the old to the recent period. Deceleration of the increasing ESRD rate from the old to the recent period was correlated with the incidence in the old period across 11 regions (r = 0.81, p < 0.003); i.e., the deceleration was greater in the regions where ESRD incidence had been higher. Whereas the increasing ESRD rate was significantly different among regions in the old period, this was not the case in the recent period, resulting in uniformity throughout Japan. CONCLUSIONS: The increasing ESRD rate is slowing in Japan, and its geographic differences, previously observed, have disappeared. PMID- 21638005 TI - Response of plasma membrane H(+)-ATPase to low temperature in cucumber roots. AB - The effect of low temperature (LT, 10 degrees C) on modification of plasma membrane (PM) H(+)-ATPase (EC 3.6.3.14) activity in cucumber roots was studied. Plants were grown under LT for 3 or 6 days. Some of the plants after 3 days exposure to LT were transferred to control conditions for another 3 days (post cold, PC). The activity of PM-H(+)-ATPase was decreased in plants treated for 3 days with LT. However, the activity of PM-H(+)-ATPase was higher in plants treated with LT for a longer time and in PC plants as well. Estimation of transcript levels of cucumber PM-H(+)-ATPase in roots indicates that the action of LT involves the gene expression level. The level of PM-H(+)-ATPase mRNA was markedly decreased in roots exposed to LT for 3 days. Moreover, the increased H(+)-ATPase activity in PM isolated from plants treated for 6 days with LT and from PC plants was positively correlated with higher levels of CsHA transcripts. Western blot analysis with an anti-phosphothreonine antibody showed that modification of the activity of PM-H(+)-ATPase under LT stress did not result from phosphorylation/dephosphorylation of the enzyme protein. However, the stimulation of PM-H(+)-ATPase activity in the case of PC plants could partially have emanated from increased activity of PM NAD(P)H oxidoreductase. In addition, modification of the transcript level of proton pump genes could have resulted from the action of H(2)O(2). In PC plants, an increase in H(2)O(2) level was observed. Moreover, treatment of plants with H(2)O(2) induced expression of PM H(+)-ATPase genes. PMID- 21638006 TI - Biomass and diversity of dry alpine plant communities along altitudinal gradients in the Himalayas. AB - A non-linear relationship between phytodiversity and altitude has widely been reported, but the relationship between phytomass and altitude remains little understood. We examined the phytomass and diversity of vascular plants along altitudinal gradients on the dry alpine rangelands of Ladakh, western Himalaya. We used generalized linear and generalized additive models to assess the relationship between these vegetation parameters and altitude. We found a hump shaped relationship between aboveground phytomass and altitude. We suspect that this is engendered by low rainfall and trampling/excessive grazing at lower slopes by domestic livestock, and low temperature and low nutrient levels at higher slopes. We also found a unimodal relationship between plant species richness and altitude at a single mountain as well as at the scale of entire Ladakh. The species-richness at the single mountain peaked between 5,000 and 5,200 m, while it peaked between 3,500 and 4,000 m at entire Ladakh level. Perhaps biotic factors such as grazing and precipitation are, respectively, important in generating this pattern at the single mountain and entire Ladakh. PMID- 21638007 TI - Pre-anaesthetic metomidate sedation delays the stress response after caudal artery cannulation in Atlantic cod (Gadus morhua). AB - Recovery from caudal artery cannulation with and without pre-anaesthesia metomidate sedation was assessed in Atlantic cod (Gadus morhua). The levels of plasma cortisol, glucose, electrolytes and acid-base parameters were compared between sedated and unsedated cod and to those in uncannulated individuals, where the samples were obtained by sacrificial sampling (reference level). Metomidate sedation delayed the stress response, causing sedated cod plasma cortisol to return to the reference level more slowly [day 4 post surgery (PS)] than in unsedated cod (day 2 PS). Plasma glucose was elevated in both sedated and unsedated cod up to and including day 5 PS. Plasma K(+) was lower and pH was higher in cannulated cod than in the reference from 24 h PS until the end of experimentation, indicating a stress effect of sacrificial sampling on plasma K(+) and pH that was likely caused by an acute stress response. Metomidate sedation delayed the stress response following CA cannulation and should therefore not be used as a pre-anaesthetic sedation in Atlantic cod. The caudal artery cannulation can be a useful tool in obtaining repeated blood samples from Atlantic cod given an adequate recovery time, which was determined to be 6 days irrespective of pre-anaesthesia sedation status. PMID- 21638008 TI - Evidence that elevated water temperature affects the reproductive physiology of the European bullhead Cottus gobio. AB - Climate change is predicted to increase the average water temperature and alter the ecology and physiology of several organisms including fish species. To examine the effects of increased water temperature on freshwater fish reproduction, adult European bullhead Cottus gobio of both genders were maintained under three temperature regimes (T1: 6-10, T2: 10-14 and T3: 14-18 degrees C) and assessed for gonad development (gonadosomatic index-GSI and gonad histology), sex steroids (testosterone-T, 17beta-estradiol-E2 and 11 ketotestosterone-11-KT) and vitellogenin (alkali-labile phosphoprotein phosphorus ALP) dynamics in December, January, February and March. The results indicate that a 8 degrees C rise in water temperature (T3) deeply disrupted the gonadal maturation in both genders. This observation was associated with the absence of GSI peak from January to March, and low levels of plasma sex steroids compared with T1-exposed fish. Nevertheless, exposure to an increasing temperature of 4 degrees C (T2) appeared to accelerate oogenesis with an early peak value in GSI and level of plasma T recorded in January relative to T1-exposed females. In males, the low GSI, reduced level of plasma 11-KT and the absence of GSI increase from January to March support the deleterious effects of increasing water temperature on spermatogenesis. The findings of the present study suggest that exposure to elevated temperatures within the context of climate warming might affect the reproductive success of C. gobio. Specifically, a 4 degrees C rise in water temperature affects gametogenesis by advancing the spawning, and a complete reproductive failure is observed at an elevated temperature of 8 degrees C. PMID- 21638009 TI - Effects of thermal acclimation on tissue fatty acid composition of freshwater alewives (Alosa pseudoharengus). AB - In this study, we examine changes in fatty acid composition of polar and neutral lipids in gill, liver, and muscle of freshwater alewives (Alosa pseudoharengus) after temperature challenges in the laboratory. Alewives experienced either a warm or cold challenge in which temperatures were raised or lowered 0.5 degrees C day(-1) over 4-6 weeks. In alewives experiencing the cold challenge, gill polar lipids showed evidence of significant remodeling, including decreases in palmitic acid and saturated fatty acids and increases in n-3 and n-6 highly unsaturated fatty acids including docosahexaenoic acid (DHA) and arachidonic acid. In alewives experiencing the warm challenge, we observed significant increases in saturated fatty acids (due mainly to increased palmitic acid) and decreases in polyunsaturated fatty acids in polar lipids of muscle and liver tissue. Fish that died during the cold challenge had significantly higher levels of palmitic acid in muscle polar lipids compared to fish that survived; fish that died during the warm challenge displayed complex changes in fatty acid composition. Based on theoretical considerations, the changes in polar lipids we observed during thermal acclimation are likely to promote appropriate membrane fluidity under each thermal regime. The increased incorporation of highly unsaturated fatty acids during cold acclimation could have significant physiological and ecological implications. In particular, since highly unsaturated fatty acids are typically scarce in freshwater food webs, dietary deficiencies in these essential fatty acids may be a significant factor in winter mortality of freshwater alewives. PMID- 21638010 TI - Mammary analogue secretory carcinoma: a new twist to the diagnostic dilemma of zymogen granule poor acinic cell carcinoma. PMID- 21638011 TI - Bmi-1, c-myc, and Snail expression in primary breast cancers and their metastases -elevated Bmi-1 expression in late breast cancer relapses. AB - Breast cancer is known for its propensity to recur decades after treatment. The biology behind the phenomenon of tumor dormancy is still poorly understood. Bmi 1, c-myc, and Snail are transcription factors that have prognostic roles in several malignancies. In order to reveal whether any of these markers has impact on late relapses, we used immunohistochemistry to study the expression of Bmi-1, c-myc, Snail, and estrogen receptor in 73 primary breast cancers and in their metastatic relapses detected within 2 years, or 5 or 10 years after primary surgery. The expression of Bmi-1 was higher in the metastases than in their corresponding primary tumors in both early and late relapses. The highest expression of Bmi-1 was seen in the very late relapsing tumors (first tumor relapse after 10 years). Previously, Bmi-1 has been reported to function as a marker of tumor stem cells in breast cancer. Our results indicate that metastases, when compared to primary tumors, arise from tumor cells that have retained stem cell properties. We also analyzed the relationship between the expression of these markers and clinical parameters. A significant association between the expression of Bmi-1 and estrogen receptor was found. Nuclear expression of c-myc in primary tumors correlated with an increased risk for axillary lymph node metastasis. PMID- 21638012 TI - In response--a modified staining protocol for Smoothelin immunostaining. PMID- 21638013 TI - Is the cerebellum a potential target for stimulation in Parkinson's disease? Results of 1-Hz rTMS on upper limb motor tasks. AB - The aim of this study was to find whether 1-Hz cerebellar repetitive transcranial magnetic stimulation (rTMS) could affect upper limb movement in early-stage Parkinson's disease (PD). Twenty patients with PD underwent one session with real and one with sham rTMS. rTMS (1 Hz, 600 pulses) was targeted at the right lateral cerebellum. Before and after rTMS, patients performed two motor tests with their fingers and hands (ball test, nine-hole peg test). The duration of these tests was measured. There were statistically significant differences (p < 0.05) in the results of the tests after real stimulation and sham stimulation. We excluded the impact of learning. After real rTMS, we observed a significantly faster response in the ball test and a slower response in the nine-hole peg test, both on the right upper limb. This study indicates the influence of 1-Hz cerebellar rTMS in modifying the voluntary movements of the upper limb in PD. This influence is differentiated: the improvement of gross motor skills and the worsening of fine motor skills. PMID- 21638014 TI - Usefulness of enhanced power Doppler imaging in monitoring acral microcirculation in type 2 diabetes mellitus and its complications. AB - This study compared hemodynamic changes of acral arterioles (pulps and nail beds of fingers and toes) and the microcirculatory status of acra between patients with uncomplicated (n = 45) or complicated (n = 36) type 2 diabetic mellitus (type 2 DM) and healthy subjects (n = 40). Enhanced power Doppler imaging (e Flow) was used to display the nail bed arterioles and distal branches of pulp arterioles (digitales palmares propriea and digitales plantares propriea) in the end knuckle of the right middle finger and right big toe. Arteriolar density (AD) was assessed by vascular pixel percentage. Compared to healthy subjects, in patients with DM the end diastolic velocity (EDV) of the nail bed arterioles of both finger and toe was diminished, while the vascular resistance index (RI) was increased. These changes became more prominent with a longer duration of the disease. Furthermore, both the peak systolic velocity (PSV) and AD were decreased in patients with DM. These hemodynamic changes were also evident in the pulp arterioles of fingers and toes, although they appeared at more advanced stages of the disease. Overall, the abnormal changes were more pronounced in patients with complications. In conclusion, hemodynamic changes (e.g. decrease in the number of acral arterioles) progress with a longer duration of the disease. The acral arteriolar damage is more pronounced in patients with a complicated type 2 DM. PMID- 21638015 TI - How uniform is the peptide plane geometry? A high-accuracy NMR study of dipolar Calpha-C'/H N-N cross-correlated relaxation. AB - Highly precise and accurate measurements of very small NMR cross-correlated relaxation rates, namely those between protein H (i) (N) -N(i) and C (i-1) (alpha) -C(i-1)' dipoles, are demonstrated with an error of 0.03 s(-1) for GB3. Because the projection angles between the two dipole vectors are very close to the magic angle the rates range only from -0.2 to +0.2 s(-1). Small changes of the average vector orientations have a dramatic impact on the relative values. The rates suggest deviation from idealized peptide plane geometry caused by twists around the C'-N bonds and/or pyramidalization of the nitrogen atoms. A clear alternating pattern along the sequence is observed in beta strands 1, 3 and 4 of GB3, where the side chains of almost all residues with large positive rates are solvent exposed. In the alpha helix all rates are relatively large and positive. Some of the currently most accurate structures of GB3 determined by both high resolution X-ray crystallography and NMR are in satisfactory agreement with the experimental rates in the helix and beta strand 3, but not in the loops and the two central strands of the sheet for which no alternating pattern is predicted. PMID- 21638016 TI - Novel heterozygous tissue-nonspecific alkaline phosphatase (TNAP) gene mutations causing lethal perinatal hypophosphatasia. AB - Hypophosphatasia is a rare inherited disorder characterized by poor bone mineralization and deficiency of alkaline phosphatase activity. It is caused by mutations in the liver/bone/kidney alkaline phosphatase gene encoding the tissue nonspecific isoenzyme of alkaline phosphatase (TNAP), which displays many allelic heterogeneities, leading to different clinical phenotypes. This study reports the case of a patient diagnosed with lethal perinatal hypophosphatasia. His gene analysis showed compound heterozygocity of two novel mutations: c.650delTinsCTAA and c.984_986delCTT, which led to p.217delVinsAK and p.328delF, respectively. The two mutations originated separately from his parents, consistent with autosomal recessive perinatal hypophosphatasia. For these two novel mutations, we analyzed their functions through three-dimensional structural analysis. This revealed that V217 is located in the beta-sheet area, V217 is deleted, and insertion of alanine and lysine alter the secondary structure, causing instability in the hydrophobic region, which may influence the metal-binding vicinity. This mutant structure loses its catalytic activity. Deletion of 328F also results in protein structural alteration and affects TNAP functions. These results may provide an explanation of the two novel mutated alleles correlating with the lethal phenotype of our patient. In conclusion, we demonstrated the case of a patient with lethal perinatal hypophosphatasia caused by two novel heterozygous mutations. PMID- 21638017 TI - Soy protein diet and exercise training increase relative bone volume and enhance bone microarchitecture in a mouse model of uremia. AB - Soy protein consumption and exercise training have been widely studied for their effects on the vasculature and bone in healthy populations, but little is known about the effectiveness of these interventions in chronic kidney disease (CKD). Cardiovascular disease and bone fracture risk are significantly elevated in CKD, and current pharmacological interventions have been unsuccessful in treating these conditions simultaneously. The purpose of this study was to compare the effects of a soy protein diet and endurance exercise training, alone or in combination, on cardiovascular and bone health in a mouse model of renal insufficiency. At 8 weeks of age, 60 female apolipoprotein E(-/-) mice underwent a two-step surgical procedure to induce uremia. These mice were then randomized at 12 weeks of age to one of four treatment groups for the 16-week intervention period: sedentary, control diet (n = 16); sedentary, soy protein diet (n = 18); exercise, control diet (n = 14); and exercise, soy protein diet (n = 12). There were no significant treatment effects on atherosclerotic lesion areas or aortic calcium deposits. We demonstrated a significant main effect of both diet and exercise on relative bone volume, trabecular number, trabecular separation, and trabecular connective density in the proximal femur as measured by microcomputed tomography. There were no treatment effects on trabecular thickness. We also showed a main effect of diet on plasma urea levels. These data suggest that soy protein intake and exercise training exert beneficial effects on properties of bone and plasma urea levels in mice with surgically induced renal impairment. PMID- 21638018 TI - Identification of genes for bone mineral density variation by computational disease gene identification strategy. AB - We previously used five freely available bioinformatics tools (Prioritizer, Geneseeker, PROSPECTR and SUSPECTS, Disease Gene Prediction, and Endeavour) to analyze the thirteen well-replicated osteoporosis susceptibility loci and identify a subset of most likely candidate osteoporosis susceptibility genes (Huang et al. in J Hum Genet 53:644-655, 2008). In the current study, we experimentally tested the association between bone mineral density (BMD) and the 9 most likely candidate genes [LAMC2(1q25-q31), MATN3(2p24-p23), ITGAV(2q31-q32), ACVR1(2q23-q24), TDGF1(3p21.31), EGF(4q25), IGF1(12q22-q23), ZIC2(13q32), BMP2(20p12)] which were pinpointed by 4 or more bioinformatics tools. Forty tag SNPs in nine candidate genes were genotyped in a southern Chinese female case control cohort consisting of 1643 subjects. Single- and multi-marker association analyses were performed using logistic regression analysis implemented by PLINK. Potential transcription factor binding sites were predicted by MatInspector. The strongest association was observed between rs10178256 (MATN3) and trochanter (P < 0.001) and total hip BMD (P = 0.002). The SNP rs6214 (IGF1) showed consistent association with BMD at all the four measured skeletal sites (P = 0.005-0.044). Prediction of transcription factor binding suggested that the minor allele G of rs10178256 might abolish the binding of MESP1 and MESP2 which play vital roles in bone homeostasis, whereas the minor allele G of rs6214 might create an additional binding site for XBP1, a constitutive regulator of endoplasmic reticulum stress response. Our data suggested that variants in MATN3 and IGF1 were involved in BMD regulation in southern Chinese women. PMID- 21638019 TI - Efficient synthesis of unnatural dipeptides based on cis-2,5-disubstituted pyrrolidine. AB - The well-defined unnatural dipeptides based on cis-2,5-disubstituted pyrrolidine backbone were synthesized from commercially available starting materials meso diethyl-2,5-dibromoadipate, (S)-(-)-1-phenylethylamine, and phenylalanine. The configurations of all the chiral centers in these unnatural dipeptides are confirmed by X-ray crystal diffraction analysis. PMID- 21638020 TI - Effects of dietary L-arginine or N-carbamylglutamate supplementation during late gestation of sows on the miR-15b/16, miR-221/222, VEGFA and eNOS expression in umbilical vein. AB - Placental vascular formation and blood flow are crucial for fetal survival, growth and development, and arginine regulates vascular development and function. This study determined the effects of dietary arginine or N-carbamylglutamate (NCG) supplementation during late gestation of sows on the microRNAs, vascular endothelial growth factor A (VEGFA) and endothelial nitric oxide synthase (eNOS) expression in umbilical vein. Twenty-seven landrace*large white sows at day (d) 90 of gestation were assigned randomly to three groups and fed the following diets: a control diet and the control diet supplemented with 1.0% L-arginine or 0.10% NCG. Umbilical vein of fetuses with body weight around 2.0 kg (oversized), 1.5 kg (normal) and 0.6 kg (intrauterine growth restriction, IUGR) were obtained immediately after farrowing for miR-15b, miR-16, miR-221, miR-222, VEGFA and eNOS real-time PCR analysis. Compared with the control diets, dietary Arg or NCG supplementation enhanced the reproductive performance of sows, significantly increased (P<0.05) plasma arginine and decreased plasma VEGF and eNOS (P<0.05). The miR-15b expression in the umbilical vein was higher (P<0.05) in the NCG supplemented group than in the control group. There was a trend in that the miR 222 expression in the umbilical vein of the oversized fetuses was higher (0.05 betagamma modification can be harnessed by reducing the solvent-exposed hydrophobic surface area and placing of suitably long and bulky helix-forming side chains at the hydrophobic core. The pairing of alphabetagamma-chimeric sequences with the native wild-type are thermodynamically allowed in the case of ideal arrangement of beta- and gamma-residues. This indicates a similarity in local side chain packing of beta- and gamma-amino acids at the helical interface of alphabetagamma-chimeras and the native alpha-peptide. Consequently, the backbone extended residues are able to participate in classical "knob-into-hole" packing with native alpha-peptide. PMID- 21638023 TI - How common is methotrexate pneumonitis? A large prospective study investigates. AB - The current literature states the prevalence of methotrexate pneumonitis (MTX-P) to be 3.5-7.6%. This is based on retrospective data. Consequently, clinicians remain cautious in using methotrexate especially in patients with pre-existing lung disease. To get a true idea of the incidence of MTX-P we designed an ongoing prospective study, which is the largest to date. We recruited all patients starting low-dose methotrexate in our department, and followed them up for 2 years or until development of MTX-P. All patients had their pulmonary spirometry checked at baseline. Patients were excluded if they did not give consent for methotrexate therapy, or had a forced expiratory volume in 1 s (fev1) or full vital capacity (FVC) of less than 1 l. So far, 223 patients have been recruited of whom 223 have completed 6 months and 185 have finished 2 years of follow-up from commencing methotrexate. Only two patients developed MTX-P. This gave an incidence of one case every 192 patient years of MTX-P. The results of this ongoing prospective study suggest that MTX-P when diagnosed using Carson's criteria and Chest HRCT scanning, does not occur as often as previously thought. Also it would appear from our data that baseline spirometry rather than full pulmonary function tests can be used routinely as an immediate screening of lung function prior to commencement of methotrexate. Interestingly the patients who developed MTX-P did not have any specific abnormalities at baseline. PMID- 21638024 TI - Identification of serum biomarkers for ovarian cancer using MALDI-TOF-MS combined with magnetic beads. AB - BACKGROUND: The objective of this study was to search for potential protein biomarkers in serum for diagnosis of ovarian cancer, by use of proteomic fingerprint techniques. METHOD: MALDI-TOF-MS was combined with magnetic beads to profile and compare serum protein spectra from 40 ovarian cancer patients and from 60 healthy controls. RESULTS: The tree analysis model of potential cancer biomarkers was constructed with Biomarker Patterns software on the basis of three identified biomarkers (5486, 6440, and 13720 Da), resulting in excellent discrimination between the ovarian cancer and non-cancer in our tests. The sensitivity was 90% and the specificity was 86.7%. In a blind test the sensitivity was 88% and the specificity was 83.3%. CONCLUSION: The results suggested that biomarkers for ovarian cancer diagnosis in serum could be identified by MALDI-TOF-MS combined with the use of magnetic beads. The use of combined biomarkers would further enable powerful and reliable diagnosis of ovarian cancer with high sensitivity and specificity. PMID- 21638025 TI - Duodenal metastasis from recurrent invasive lobular carcinoma of breast: a case report and literature review. AB - We present a rare case of duodenal metastasis from invasive breast lobular carcinoma, which first presented clinically as elevated serum tumor marker levels, followed by jaundice but with no other clinical evidence of recurrence and metastasis. A 53-year-old woman underwent modified radical mastectomy of the left breast (pT2 N3 M0 stage III c) followed by postoperative chemo-radiotherapy and hormonal therapy. After about 3 years, the patient presented with elevated serum tumor marker levels and mild jaundice. She was subsequently admitted to the hospital for nausea and severe vomiting. A duodenoscopy revealed the thickening of duodenal papilla on the lateral wall and stenosis. A duodenal tissue biopsy revealed poorly differentiated adenocarcinoma, and immunohistochemical staining suggested that the carcinoma was of breast origin. The patient received further radiation and chemotherapy. Although duodenal metastases of breast cancer are rare, physicans should be alert and vigilant when a patient with a history of breast cancer presents with new gastrointestinal symptoms. PMID- 21638026 TI - Alternative promoter usage and differential expression of multiple transcripts of mouse Prkar1a gene. AB - Prkar1a gene encodes regulatory type 1 alpha subunit (RIalpha) of cAMP-dependent protein kinase (PKA) in mouse. The role of this gene has been implicated in Carney complex and many cancer types that suggest its involvement in physiological processes like cell cycle regulation, growth and/or proliferation. We have identified and sequenced partial cDNA clones encoding four alternatively spliced transcripts of mouse Prkar1a gene. These transcripts have alternate 5' UTR structure which results from splicing of three exons (designated as E1a, E1b, and E1c) to canonical exon 2. The designated transcripts T1, T2, T3, and T4 contain 5' UTR exons as E1c, E1a + E1b, E1a, and E1b, respectively. The transcript T1 corresponded to earlier reported transcript in GenBank. In silico study of genomic DNA sequence revealed three distinct promoter regions namely, P1, P2, and P3 upstream of the exons E1a, E1b, and E1c, respectively. P1 is non CpG-related promoter but P2 and P3 are CpG-related promoters; however, all three are TATA less. RT-PCR analysis demonstrated the expression of all four transcripts in late postnatal stages; however, these were differentially regulated in early postnatal stages of 0.5 day, 3 day, and 15 day mice in different tissue types. Variations in expression of Prkar1a gene transcripts suggest their regulation from multiple promoters that respond to a variety of signals arising in or out of the cell in tissue and developmental stage-specific manner. PMID- 21638028 TI - Novel screening cascade identifies MKK4 as key kinase regulating Tau phosphorylation at Ser422. AB - Phosphorylation of Tau at serine 422 promotes Tau aggregation. The kinase that is responsible for this key phosphorylation event has so far not been identified but could be a potential drug target for Alzheimer's disease. We describe here an assay strategy to identify this kinase. Using a combination of screening a library of 65'000 kinase inhibitors and in vitro inhibitor target profiling of the screening hits using the Ambit kinase platform, MKK4 was identified as playing a key role in Tau-S422 phosphorylation in human neuroblastoma cells. PMID- 21638027 TI - Dual role of insulin-like growth factor-1 in acetyl-CoA carboxylase-alpha activity in human colon cancer cells HCT-8: downregulating its expression and phosphorylation. AB - Insulin-like growth factor-1 (IGF-1) plays the role in cellular lipid synthesis and cell proliferation. However, the role of IGF-1 on the growth of colon cancer cell line HCT-8 is not clear. In this study, HCT-8 cells were exposed to IGF-1 at 0, 10, 50, or 100 ng/ml in serum-free medium. Fatty acid/lipid synthesis in HCT-8 cells was examined by 2-14C-acetate incorporation. HCT-8 cell growth and proliferation were determined by MTT assay and Trypan blue exclusive viable cell counting. We found that in serum starvation conditions, IGF-1 at 10-100 ng/ml induced dose-dependent down regulation of both the ACCalpha expression and the phosphorylation in HCT-8 cells, maintaining a balance in ACCalpha activity and lipid synthesis. IGF-1 reduced p-ATM, p-AMPK, and then p-ACCalpha protein levels in HCT-8 cells. IGF-1 increased p-Akt levels, but decreased p-ERK1/2 levels, leading to the decrease in ACCalpha protein and mRNA levels. Similarly, ERK1/2 inhibitor PD98059 reduced ACCalpha expression. IGF-1 influences neither HCT-8 cell growth nor their p53 protein levels and PARP cleavage. In a word, IGF-1 reduced ACCalpha phosphorylation via an ATM/AMPK signaling pathway and suppressed ACCalpha expression through an ERK1/2 transduction, playing a dual role in regulating ACCalpha activity and lipogenesis. This may render a cell with survival advantages under a serum starvation crisis, representing a novel mitogenic role of IGF-1. PMID- 21638029 TI - Clavulanic acid does not affect convulsions in acute seizure tests in mice. AB - Clavulanic acid (CLAV) inhibits bacterial beta-lactamases and is commonly used to aid antibiotic therapy. Prompted by the initial evidence suggestive of the potential anticonvulsant and neuroprotective properties of CLAV, the present study was undertaken to systematically evaluate its acute effects on seizure thresholds in seizure tests typically used in primary screening of potential antiepileptic drugs (AEDs). In the present study, 6-Hz seizure threshold, maximal electroshock seizure threshold (MEST) test, and intravenous pentylenetetrazole (i.v. PTZ) seizure tests were used to determine anticonvulsant effects of intraperitoneally (i.p.) administered CLAV in mice. Acute effects on motor coordination and muscle strength were assessed in the chimney and grip-strength tests, respectively. Doses of CLAV studied in the present study were either comparable or extended the doses reported in the literature to be effective against kainic acid-induced convulsions in mice or behaviorally active in rodents and monkeys. CLAV had no effect on seizure thresholds in the 6-Hz (64 ng/kg to 1 mg/kg) and MEST (64 ng/kg to 5 mg/kg) seizure tests. Similarly, CLAV had no effect on seizure thresholds for i.v. PTZ-induced myoclonic twitch, clonic convulsions, and tonic convulsions (64 ng/kg to 5 mg/kg). Finally, CLAV (64 ng/kg to 5 mg/kg) had no effect on the motor performance and muscle strength in the chimney and grip-strength tests, respectively. In summary, CLAV failed to affect seizure thresholds in three seizure tests in mice. Although the results of the present study do not support further development of CLAV as an AED, its beneficial effects in chronic epilepsy models warrant further evaluation owing to its, for example, potential neuroprotective properties. PMID- 21638030 TI - Optic disc pit: a review. AB - BACKGROUND: Since Wiethe first described the clinical presentation of two optic disc depressions in a 62-year-old woman in 1882, there have been many studies addressing what later become known as the "optic disc pit." The main complication of this condition, termed optic disc pit maculopathy, is associated with visual deterioration. Treatment of optic disc pit maculopathy remains challenging. METHODS: Here we review the body of literature that documents the clinical findings, pathophysiology, histology, main complications, treatment options, special features and presentations, and differential diagnosis of optic disc pit. RESULTS: The source of the intraretinal fluid in optic disc pit maculopathy remains controversial. Four possible sources of this fluid have been proposed: fluid from the vitreous cavity; cerebrospinal fluid originating from the subarachnoid space; fluid from leaky blood vessels at the base of the pit; and fluid from the orbital space surrounding the dura. CONCLUSIONS: Optic disc pits are a very rare clinical entity, affecting approximately one in 11,000 people. Patients with congenital optic disc pit sometimes remain asymptomatic, but 25% to 75% present with visual deterioration in their 30s or 40s after developing macular schisis and detachment. The most widely accepted treatment for such patients is a surgical approach involving pars plana vitrectomy with or without internal limiting membrane peeling, with or without endolaser photocoagulation and C3F8 endotamponade. PMID- 21638031 TI - Unsuccessful surgical excision of optic nerve drusen. AB - BACKGROUND: To report the case of failed surgical excision of optic nerve drusen (OND). METHODS: Case report. A 53-year-old woman presented with bilateral OND leading to progressive field defects and LE light perception and RE 20/25 vision. A vitrectomy was performed on the legally-blind left eye to test the resectability of OND. RESULTS: The removal of superficial OND failed because the singular-appearing superficial OND presented as one large mass with multiple excrescences on its surface. Retinal vessels passed through it, which made an excision impossible. CONCLUSION: OND cannot always be removed surgically, due to their variable consistency and hardness. PMID- 21638032 TI - A case of haemorrhagic cystitis by inhaled salbutamol and salmeterol. PMID- 21638033 TI - Pharmacokinetics of caspofungin in a critically ill patient with liver cirrhosis. PMID- 21638034 TI - Pelvic ultrasound immediately following MDCT in female patients with abdominal/pelvic pain: is it always necessary? AB - To determine the added value of reimaging the female pelvis with ultrasound (US) immediately following multidetector CT (MDCT) in the emergent setting. CT and US exams of 70 patients who underwent MDCT for evaluation of abdominal/pelvic pain followed by pelvic ultrasound within 48 h were retrospectively reviewed by three readers. Initially, only the CT images were reviewed followed by evaluation of CT images in conjunction with US images. Diagnostic confidence was recorded for each reading and an exact Wilcoxon signed rank test was performed to compare the two. Changes in diagnosis based on combined CT and US readings versus CT readings alone were identified. Confidence intervals (95%) were derived for the percentage of times US reimaging can be expected to lead to a change in diagnosis relative to the diagnosis based on CT interpretation alone. Ultrasound changed the diagnosis for the ovaries/adnexa 8.1% of the time (three reader average); the majority being cases of a suspected CT abnormality found to be normal on US. Ultrasound changed the diagnosis for the uterus 11.9% of the time (three reader average); the majority related to the endometrial canal. The 95% confidence intervals for the ovaries/adnexa and uterus were 5-12.5% and 8-17%, respectively. Ten cases of a normal CT were followed by a normal US with 100% agreement across all three readers. Experienced readers correctly diagnosed ruptured ovarian cysts and tubo-ovarian abscesses (TOA) based on CT alone with 100% agreement. US reimaging after MDCT of the abdomen and pelvis is not helpful: (1) following a normal CT of the pelvic organs or (2) when CT findings are diagnostic and/or characteristic of certain entities such as ruptured cysts and TOA. Reimaging with ultrasound is warranted for (1) less-experienced readers to improve diagnostic confidence or when CT findings are not definitive, (2) further evaluation of suspected endometrial abnormalities. A distinction should be made between the need for immediate vs. follow-up imaging with US after CT. PMID- 21638035 TI - Expression and functional characterization of P2X receptors in mouse alveolar macrophages. AB - Alveolar macrophages (AM) are crucial for pulmonary host defense, and evidence emerges that ATP-gated P2X receptors are involved in inflammatory processes. This study focuses on the expression and functional characterization of P2X receptors in AM from mouse. In RT-PCR experiments, transcripts encoding the P2X1, P2X3, P2X4, P2X5, and P2X7 receptors were detected. In whole-cell patch-clamp recordings, ATP (1 mM) evoked an inward current (mouse and human AM) that was reversible upon washout, and the reversal potential was ~5 mV, indicating the activation of a non-selective conductance-a fingerprint of P2X receptors. Further characterization (mouse AM) revealed that the current was not desensitized by a second ATP application. The ATP-induced current was increased by the removal of extracellular Ca2+ (in human and mouse AM), and EC50 in mouse AM were determined with ~1 mM ATP, in the presence as well as in the absence of extracellular Ca2+. Pharmacological characterization of mouse AM revealed that the effect was augmented by BzATP and pre-application with ivermectin, but no effect with alpha,beta-meATP was observed. Further, the ATP effect was reduced by PPADS (300 MUM), brilliant blue G (5 MUM), and about A438079 (10 MUM). Although different P2X receptor transcripts were detected in mouse AM, the observed functional and pharmacological characteristics indicate primarily the participation of P2X4 and P2X7 receptors as mediators of the ATP-induced ion current in mouse AM. These suggestions were confirmed by experiments with AM from P2X7 -deficient animals, indicating a contribution of P2X4 and P2X7 receptors in pulmonary immune function. PMID- 21638036 TI - GmFtsH9 expression correlates with in vivo photosystem II function: chlorophyll a fluorescence transient analysis and eQTL mapping in soybean. AB - Filamentation temperature-sensitive H (FtsH) is an ATP-dependent zinc metalloprotease involved in diverse biological functions. There are 12 FtsH proteins in Arabidopsis, among which AtFtsH2 plays an important role in regulating the turnover of photosystem II (PSII) reaction center D1 protein and the development of the photosynthetic apparatus. Here, we have identified 11 FtsH genes in the soybean genome by a bioinformatics approach. These soybean FtsH genes corresponded to seven Arabidopsis FtsH genes, suggesting that the main characteristics of soybean FtsH genes were formed before the evolutionary split of soybean and Arabidopsis. Phylogenetic analyses allowed us to clone a soybean AtFtsH2-like gene designated as GmFtsH9. The predicted protein of GmFtsH9 consists of 690 amino acids and contains three typical FtsH proteins conserved domains. The expression level of GmFtsH9 was determined in a soybean recombinant inbred line population under a pot experiment conducted for measuring chlorophyll a fluorescence transient parameters, photosynthetic CO(2) fixation rate (P (N)), and seed yield. Expression quantitative trait loci (eQTL) mapping revealed two trans-acting eQTLs for GmFtsH9. The significant correlation of gene expression level with chlorophyll a fluorescence transient parameters and the presence of overlapping eQTL (QTL) between gene expression level and chlorophyll a fluorescence transient parameters indicated that GmFtsH9 could be involved in regulating PSII function. These results further lead to the understanding of the mechanism underlying FtsH gene expression, and contribute to the development of marker-assisted selection breeding programs for modulating soybean FtsH gene expression. PMID- 21638037 TI - Protective effect of propofol and its relation to postoperation recovery in children undergoing cardiac surgery with cardiopulmonary bypass. AB - The aim of this study was to investigate the effect of propofol and its relation to postoperation recovery in children undergoing cardiac surgery with cardiopulmonary bypass (CPB). Twenty ASA class I-II children with congenital heart disease undergoing cardiac surgery were randomly allocated to a propofol group (n = 10) or a control group (n = 10). Blood samples were collected at five time points: before operation (T (0)), before the start of CPB (T (1)), 25 min after the aorta was cross-clamped (T (2)), 30 min after release of the aortic cross-clamp (T (3)), and 2 h after the cessation of CPB (T (4)). The myocardial samples were collected at the time of incubation into the right atrium before CPB and at 30 min after reperfusion. After CPB, propofol significantly suppressed the increase of the serum lactate dehydrogenase (LDH), creatine phosphokinase (CK), and interleukin-6 (IL-6) levels and the decrease of the serum superoxide dismutase (SOD) level. In addition, propofol inhibited the increase of myocardial nuclear factor-kappaB (NF-kappaB) expression and inflammatory cells infiltration after CPB. Furthermore, propofol significantly shortened the tracheal extubation time. In conclusion, propofol exerts a protective effect and improves postoperation recovery through its antioxidant and anti-inflammatory actions in children undergoing cardiac surgery with CPB. PMID- 21638038 TI - Right coronary artery fistula to the right atrium. PMID- 21638039 TI - Central fat influences cardiac autonomic function in obese and overweight girls. AB - It has been suggested that upper-body fat compared with lower-body fat is more closely associated with cardiovascular abnormalities. Our objective was to analyze the relationship between central fat (CF) and cardiac autonomic (cANS) function in obese and overweight girls. Children were classified in two groups based on CF: those above (CFa(50)) and those below the 50th percentile (CFb(50)) of the entire sample. This study included 16 female children who were diagnosed as being overweight or obese (age: 14.3 +/- 2.8 years; weight: 75.0 +/- 15.8 kg; height: 157.1 +/- 8.9 cm; body mass index: 30.1 +/- 5.4; and total body fat: 40.5 +/- 5.0%; Tanner stage: 4). cANS function was assessed through heart rate variability (HRV) and CF parameters by dual-energy X-ray absorptiometry. Female children with higher CF exhibited significantly higher sympathetic and lower parasympathetic modulation than those with lower CF, independently of total body fat. The data of the present study indicate that CF is associated with less favorable indexes of HRV. In addition, our findings suggest that CF might be an important measure to assess the effect of obesity on cANS function in female children. PMID- 21638040 TI - The community dynamics of major bioleaching microorganisms during chalcopyrite leaching under the effect of organics. AB - To determine the effect of organics (yeast extract) on microbial community during chalcopyrite bioleaching at different temperature, real-time polymerase chain reaction (PCR) was employed to analyze community dynamics of major bacteria applied in bioleaching. The results showed that yeast extract exerted great impact on microbial community, and therefore influencing bioleaching rate. To be specific, yeast extract was adverse to this bioleaching process at 30 degrees C due to decreased proportion of important chemolithotrophs such as Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans. However, yeast extract could promote bioleaching rate at 40 degrees C on account of the increased number and enhanced work of Ferroplasma thermophilum, a kind of facultative bacteria. Similarly, bioleaching rate was enhanced under the effect of yeast extract at 50 degrees C owing to the work of Acidianus brierleyi. At 60 degrees C, bioleaching rate was close to 100% and temperature was the dominant factor determining bioleaching rate. Interestingly, the existence of yeast extract greatly enhanced the relative competitiveness of Ferroplasma thermophilum in this complex bioleaching microbial community. PMID- 21638041 TI - Genetic and electron-microscopic characterization of 'Rickettsiella agriotidis', a new Rickettsiella pathotype associated with wireworm, Agriotes sp. (Coleoptera: Elateridae). AB - Wireworms, the polyphagous larvae of click beetles belonging to the genus Agriotes (Coleoptera: Elateridae), are severe and widespread agricultural pests affecting numerous crops. A previously unknown intracellular bacterium has been identified in a diseased Agriotes larva. Microscopic studies revealed the subcellular structures characteristic of Rickettsiella infections. Molecular phylogenetic analysis based on 16S ribosomal RNA and signal recognition particle receptor (FtsY) encoding sequences demonstrates that the wireworm pathogen belongs to the taxonomic genus Rickettsiella. Therefore, the new pathotype designation 'R. agriotidis' is proposed to refer to this organism. Moreover, genetic analysis makes it likely that--on the basis of the currently accepted organization of the genus Rickettsiella--this new pathotype should be considered a synonym of the nomenclatural type species, Rickettsiella popilliae. PMID- 21638042 TI - The green microalga Chlorella saccharophila as a suitable source of oil for biodiesel production. AB - The aim of this study was to investigate the potential of the green microalga Chlorella saccharophila as a source of oil for biodiesel production. We evaluated for the first time, the effect of salinity and/or nitrogen depletion (ND) on cell growth, lipid accumulation and lipid profile in this microalga. The fatty acid methyl esters (FAME) identified for C. saccharophila in this study consisted of C 16:0, C-18:0, C-18:1 cis, and C-18:1 trans. Among these, C-18:1 (indicator of biodiesel quality) was the main FAME found, representing approximately 76 and 80% of total FAME under normal and ND growing conditions, respectively. Under a normal growing condition this microalga showed 154.63 mg l(-1) d(-1), 63.33 mg l( 1) d(-1), and 103.73 mg l(-1) of biomass productivity, lipid productivity, and FAME yield, respectively. The higher biomass productivity (159.58 mg l(-1) d( 1)), lipid productivity (99.33 mg l(-1) d(-1)), and FAME yield (315.53 mg l(-1)) were obtained under the ND treatment. In comparison to other related studies, our results suggest that C. saccharophila can be considered as a suitable source of oil for biodiesel production. PMID- 21638043 TI - Expression, characterization, and site-directed mutation of a multiple herbicide resistant acetohydroxyacid synthase (rAHAS) from Pseudomonas sp. Lm10. AB - A multiple herbicide-resistant acetohydroxyacid synthase (rAHAS) gene was cloned from Pseudomonas sp. Lm10. Sequence analysis showed that the rAHAS regulatory subunit was identical to that of Pseudomonas putida KT2440 (sensitive AHAS, sAHAS), whereas six different sites [H134->N (rAHAS->sAHAS), A135->P, S136->T, I210->V, F264->Y, and S486->W] were found in the catalytic subunit. The rAHAS and sAHAS were over expressed, purified and characterized. rAHAS showed higher resistance to four kinds of AHAS-inhibitor herbicides than sAHAS. The resistance factor of rAHAS was 56.0-fold, 12.6-fold, 6.5-fold, and 9.2-fold as compared with sAHAS when metsulfuron-methyl, imazethapyr, flumetsulam, and pyriminobac-methyl used as inhibitor, respectively. The specific activity of rAHAS was lower than that of sAHAS and the K (m) value of rAHAS for pyruvate was approximately onefold higher than the corresponding value for sAHAS. Data from site-directed mutagenesis demonstrated that alteration at A135, F264, and S486 resulted in resistance reduction, while the mutation at H134, S136, and I210 has little effect on the resistance. A135 was mainly responsible for resistance to imidazolinone; F264 conferred resistance to sulfonylurea and triazolopyrimidine sulfonamide; and S486 showed multiple herbicides resistance to the four herbicides. PMID- 21638044 TI - Time-dependent density functional theory study on the electronic excited-state hydrogen bonding of the chromophore coumarin 153 in a room-temperature ionic liquid. AB - In the present work, in order to investigate the electronic excited-state intermolecular hydrogen bonding between the chromophore coumarin 153 (C153) and the room-temperature ionic liquid N,N-dimethylethanolammonium formate (DAF), both the geometric structures and the infrared spectra of the hydrogen-bonded complex C153-DAF(+) in the excited state were studied by a time-dependent density functional theory (TDDFT) method. We theoretically demonstrated that the intermolecular hydrogen bond C(1) = O(1)...H(1)-O(3) in the hydrogen-bonded C153 DAF(+) complex is significantly strengthened in the S(1) state by monitoring the spectral shifts of the C=O group and O-H group involved in the hydrogen bond C(1) = O(1)...H(1)-O(3). Moreover, the length of the hydrogen bond C(1) = O(1)...H(1) O(3) between the oxygen atom and hydrogen atom decreased from 1.693 A to 1.633 A upon photoexcitation. This was also confirmed by the increase in the hydrogen bond binding energy from 69.92 kJ mol(-1) in the ground state to 90.17 kJ mol(-1) in the excited state. Thus, the excited-state hydrogen-bond strengthening of the coumarin chromophore in an ionic liquid has been demonstrated theoretically for the first time. PMID- 21638045 TI - Insights into the permeability of drugs and drug-like molecules from MI-QSAR and HQSAR studies. AB - Membrane-interaction QSAR (MI-QSAR) and Holographic QSAR (HQSAR) analyses have been performed on a diverse set of drugs and drug-like molecules. MI-QSAR combines a set of membrane-solute interaction properties calculated during molecular dynamics simulation with the set of classical solute descriptors to predict the biological behavior of drugs and drug-like molecules. HQSAR is a technique which employs fragment fingerprints or molecular holograms as predictive variables of biological activity. A data set of 60 structurally diverse molecules with permeability coefficients were used to construct significant MI-QSAR and HQSAR models of Caco-2 cell permeation. A statistically meaningful MI-QSAR model was obtained with r (2) = 0.805 and q (2) = 0.696. Subsequently, HQSAR models were developed on the same data set. The best HQSAR model (r (2) = 0.915, q (2) = 0.539) was obtained with fragment distinctions atom, bond, donor and acceptor with atom count 4 to 7. The predictions for training and test set molecules are in good agreement with experimental results and show the potential of models for untested compounds. This displays the importance of MI-QSAR and HQSAR analysis in estimating ADME properties characterized by the transport of solutes through biological membranes. PMID- 21638046 TI - Invasive placentation and uterus preserving treatment modalities: a systematic review. AB - PURPOSE: We present a systematic review to evaluate failure rates (secondary hysterectomy or maternal mortality) and success rates (subsequent menstruation or pregnancy) after different uterus preserving treatment modalities in women with invasive placentation. METHODS: A review of English, German or Dutch language published research, using Medline and Embase databases, was performed. Studies of any design were included. RESULTS: Ten cohort studies and 50 case series or case reports were included. Expectant management reported a secondary hysterectomy in 55/287 (19%), maternal mortality in 1/295 (0.3%), a subsequent menstruation in 44/49 (90%) and a subsequent pregnancy in 24/36 (67%). Embolization of the uterine arteries described a secondary hysterectomy in 8/45 (18%), a subsequent menstruation in 8/13 (62%) and a subsequent pregnancy in 5/33 (15%). Methotrexate therapy presented a secondary hysterectomy in 1/16 (6%), a subsequent menstruation in 4/5 (80%) and a subsequent pregnancy in 1/2 (50%). Uterus preserving surgery showed a secondary hysterectomy in 24/77 (31%), maternal mortality in 2/55 (4%), a subsequent menstruation in 28/34 (82%) and a subsequent pregnancy in 19/26 (73%). CONCLUSIONS: This review indicates that different uterus preserving treatment modalities may be effective in managing invasive placentation. Despite the extensive review of the literature, no conclusions about the superiority of any modality can be drawn. PMID- 21638047 TI - Acute sex hormone suppression reduces skeletal muscle sympathetic nerve activity. AB - OBJECTIVES: Comparisons of sympathetic nervous system activity (SNA) between young and older women have produced equivocal results, in part due to inadequate control for potential differences in sex hormone concentrations, age, and body composition. The aim of the present study was to determine the effect of a short term reduction in sex hormones on tonic skeletal muscle sympathetic nerve activity (MSNA), an indirect measure of whole body SNA, using an experimental model of sex hormone deficiency in young women. We also assessed the independent effects of estradiol and progesterone add-back therapy on MSNA. METHODS: MSNA was measured in 9 women (30+/-2 years; mean+/-SE) on three separate occasions: during the mid-luteal menstrual cycle phase, on the fifth day of gonadotropin-releasing hormone antagonist (GnRHant) administration, and after 5 days add-back of either estradiol (n=4) or progesterone (n=3) during continued GnRHant administration. RESULTS: In response to GnRHant, there were significant reductions in serum estradiol and progesterone (both p<0.01) and MSNA (25.0+/-1.9 vs. 19.2+/-2.4 bursts/min, p=0.04). Continued GnRHant plus add-back estradiol or progesterone resulted in a nonsignificant decrease (19.2+/-1.7 vs. 12.1+/-1.9 bursts/min, p=0.07) or increase (16.2+/-1.7 vs. 21.0+/-6.0 bursts/min, p=0.39), respectively, in MSNA when compared with GnRHant alone. INTERPRETATION: The findings of this preliminary study suggest that short-term ovarian hormone suppression attenuates MSNA and that this may be related to the suppression of progesterone rather than estradiol. PMID- 21638048 TI - Effects of toremifene and anastrozole on serum lipids and bone metabolism in postmenopausal females with estrogen receptor-positive breast cancer: the results of a 2-year multicenter open randomized study. AB - The potential long-term adverse effects on quality of life have to be considered when selecting agents for adjuvant hormonal treatment for postmenopausal patients with estrogen receptor-positive breast cancer. We performed a 2-year multicenter randomized study to assess the differences in the time course effects between toremifene (TOR) and anastrozole (ANA) on serum lipid profiles and bone metabolism. This study assessed the serum levels of triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), apolipoprotein A-1 (Apo A1), and apolipoprotein B (Apo B) as lipid profiles and bone-specific alkaline phosphatase (BAP) and the N-telopeptide of type-I collagen (NTX) as bone turnover markers in patients who received daily doses of 40 mg and 1 mg for TOR and ANA, respectively. A decreased serum level of TC, LDL-C, and Apo B was, respectively, observed at 6 months in 6.2, 12.9, and 13.8% of the patients who received TOR compared with the baseline. These decreases were maintained for at least 24 months. These lipid levels were not changed in those who received ANA. In the TOR patients, there was an increase in the serum level of HDL-C and Apo A1 at 6 months in 17.1 and 16.3%, respectively, which was maintained for at least 24 months, whereas these levels were almost stable in the patients who received ANA. Serum BAP decreased by 12.1% at 12 months and further decreased at 24 months and the serum NTX decreased by 22.0% at 6 months, which was maintained for at least 24 months in the patients who received TOR. In contrast, the serum BAP was increased by 26.0% at 6 months and by 29.2% at 12 months and the serum NTX increased by 21.3% at 24 months compared with baseline in those received ANA. However, the serum BAP increase was not significant at 24 months. TOR provides better effects than ANA in terms of lipid profiles and bone metabolism in postmenopausal females with early breast cancer. PMID- 21638049 TI - In situ detection of HER2:HER2 and HER2:HER3 protein-protein interactions demonstrates prognostic significance in early breast cancer. AB - HER2 overexpression/amplification is linked with poor prognosis in early breast cancer. Co-expression of HER2 and HER3 is associated with endocrine and chemotherapy resistance, driven not simply by expression but by signalling via HER2:HER3 or HER2:HER2 dimers. Proximity ligation assays (PLAs) detect protein protein complexes at a single-molecule level and allow study of signalling pathways in situ. A cohort of 100 tumours was analyzed by PLA, IHC and FISH. HER complexes were analyzed by PLA in a further 321 tumours from the BR9601 trial comparing cyclophosphamide, methotrexate and fluorouracil (CMF) with epirubicin followed by CMF (epi-CMF). The relationships between HER dimer expression and RFS and OS were investigated, and multivariate regression analysis identified factors influencing patient prognosis. PLA successfully and reproducibly detected HER2:HER2 and HER2:HER3 protein complexes in vivo. A significant association (P < 0.00001) was identified between HER2 homodimerization and HER2 gene amplification. Following a minimum p value approach high levels of HER2:HER2 dimers were significantly associated with reduced relapse-free (RFS; hazard ratio = 1.72, 95% confidence interval 1.15-2.56, P = 0.008) and overall survival (OS HR = 1.69 95% CI = 1.09-2.62, P = 0.019). Similarly, high levels of HER2:HER3 dimers were associated with reduced RFS (HR = 2.18, 95% CI = 1.46-3.26, P = 0.00016) and OS (HR = 2.21, 95% CI = 1.41-3.47, P = 0.001). This study demonstrates that in situ detection of HER2 and HER2:3 protein:protein complexes can be performed robustly and reproducibly in clinical specimens, provides novel prognostic information and opens a significant novel opportunity to probe the clinical impact of cellular signalling processes. PMID- 21638050 TI - MiR-28 regulates Nrf2 expression through a Keap1-independent mechanism. AB - NF-E2-related factor 2 (Nrf2) is an important transcription factor involved in antioxidant response. Nrf2 binds antioxidant response elements (ARE) within promoters of genes encoding detoxification enzymes (e.g., NAD (P) H-quinone oxidoreductase 1 (NQO1)) leading to their transcriptional activation. Nrf2 function is regulated post-translationally by its negative regulator Kelch-like ECH-associated protein 1 (Keap1) that binds Nrf2 and induces cytoplasmic Nrf2 degradation. Our present studies provide new evidence that Nrf2 expression can be regulated by a Keap1-independent mechanism. Here, we utilized breast epithelial cells to explore the impact of microRNA (miRNA) on Nrf2 expression. We found that Nrf2 mRNA levels are reversibly correlated with miR-28 expression and that ectopic expression of miR-28 alone reduces Nrf2 mRNA and protein levels. We further investigated the molecular mechanisms by which miR-28 inhibits Nrf2 mRNA expression. Initially, the ability of miR-28 to regulate the 3' untranslated region (3'UTR) of Nrf2 mRNA was evaluated via luciferase reporter assay. We observed that miR-28 reduces wild-type Nrf2 3'UTR luciferase reporter activity and this repression is eliminated upon mutation of the miR-28 targeting seed sequence within the Nrf2 3'UTR. Moreover, over-expression of miR-28 decreased endogenous Nrf2 mRNA and protein expression. We also explored the impact of miR 28 on Keap1-Nrf2 interactions and found that miR-28 over-expression does not alter Keap1 protein levels and has no effect on the interaction of Keap1 and Nrf2. Our findings, that miR-28 targets the 3'UTR of Nrf2 mRNA and decreases Nrf2 expression, suggest that this miRNA is involved in the regulation of Nrf2 expression in breast epithelial cells. PMID- 21638051 TI - Association between polymorphisms of the renin-angiotensin system genes and breast cancer risk: a meta-analysis. AB - The renin-angiotensin system (RAS) has been considered to be implicated in the development of breast cancer. However, the results are inconsistent. In this study, we conducted a meta-analysis to assess the association between four polymorphisms, including angiotensin I-converting enzyme (ACE) I/D and A240T, angiotensin II type 1 receptor (AGTR1) A1166C and angiotensinogen (AGT) M235T polymorphisms, and breast cancer risk. Published literature from PubMed, ISI web of science, and Embase databases were retrieved. All studies evaluating the association between ACE I/D, ACE A240T, AGTR1 A1166C, or AGT M235T polymorphism and breast cancer risk were included. Pooled odds ratio (OR) with 95% confidence interval (CI) was calculated using fixed- or random-effects model. Ten studies (1,650 cases and 9,283 controls) on ACE I/D polymorphism, six studies (1,316 cases and 2,632 controls) on ACE A240T polymorphism, three studies (235 cases and 601 controls) on AGTR1 A1166C polymorphism, and two studies (273 cases and 3,547 controls) on AGT M235T polymorphism were included. Overall, the meta-analysis showed no significant association between I/D or A240T polymorphism and breast cancer risk in either genetic model. Further subgroup analysis by ethnicity also revealed non-significant association in Caucasian or Asian populations except for Africans (the statistically significant association for ACE I/D or A240T polymorphism in Africans derived from only one study). A marginally significant association was observed for AGTR1 A1166C polymorphism in Caucasians (CC vs. AA: OR = 0.31, 95% CI 0.10-0.99). In addition, there was a significant association between AGT M235T polymorphism and breast cancer risk in Caucasians (OR = 1.45, 95% CI 1.12-1.88). The present meta-analysis suggested that ACE I/D and A240T polymorphisms might not be a good predictor of breast cancer risk, while AGTR1 A1166C and AGT M235T polymorphisms might be implicated in the pathogenesis of breast cancer. Given the limited sample size, the findings warrant further investigation. PMID- 21638052 TI - Characterisation of unclassified variants in the BRCA1/2 genes with a putative effect on splicing. AB - A subset of the unclassified variants (UVs) identified during genetic screening of BRCA1/2 genes may affect splicing. We assessed at RNA level the effect of four BRCA1 and ten BRCA2 UVs with a putative splice effect, as predicted in silico. The variants selected for this study were beyond the positions -1, -2 or +1, +2 from the exon, and were not previously described (n = 8) or their effect on splicing was not assessed previously (n = 6). Lymphocytes from UV carriers and healthy controls were cultured and treated with puromycin to prevent nonsense mediated mRNA decay. The relative contribution of each allele to the various transcripts was assessed using combinations of allele-specific and transcript specific primers. BRCA2 c.425G>T, c.7976+3_7976+4del and c.8754+3G>C give rise to aberrant transcripts BRCA2Delta4, BRCA2Delta17 and retention of 46nt of intron 21, respectively, and were considered pathogenic. BRCA1 c.4987-3C>G gives rise to BRCA1Delta17 that is likely pathogenic; however, residual expression of the full length transcript from the variant allele could not be excluded. BRCA1 c.692C>T, c.693G>A and BRCA2 c.6935A>T, besides expressing the full-length transcript, increased expression of BRCA1Delta11 and BRCA2Delta12, respectively. As these are naturally occurring isoforms, also observed in controls, the clinical relevance is unclear. The seven remaining UVs did not affect splicing and three intronic variants were therefore classified as neutral. In conclusion, the RNA analysis results clarified the clinical relevance of 6 of the 14 studied UVs and thereby greatly improve the genetic counselling of high-risk breast/ovarian cancer patients carrying these classified variants. PMID- 21638053 TI - The peptide-hormone glucagon-like peptide-1 activates cAMP and inhibits growth of breast cancer cells. AB - The incretin hormone glucagon-like peptide (GLP)-1 is secreted from intestinal L cells in response to food intake, and promotes insulin secretion and pancreatic beta-cell proliferation. Reduced GLP-1 levels are observed in obesity and type 2 diabetes mellitus (T2DM) and are associated with reduced insulin secretion and increased insulin resistance. GLP-1 mediates its activities through activation of a G-protein coupled receptor, which is expressed in the pancreas, as well as other tissues. Long-acting GLP-1 receptor (GLP-1R) agonists, such as exendin-4, are currently approved for the treatment of T2DM. As obesity and T2DM are associated with increased risk of breast cancer, we aimed to explore the effects of GLP-1 and exendin-4, on breast cancer cells. Treatment with GLP-1 or exendin-4 reduced viability and enhanced apoptosis of breast cancer cells but did not affect viability of nontumorigenic cells. Moreover, exendin-4 attenuated tumor formation by breast cancer cells in athymic mice. Treatment with either GLP-1 or exendin-4 elevated cAMP levels, activated the down-stream target CREB, and enhanced CRE promoter transcription, in breast cancer cells. Moreover, inhibition of exendin-4-induced adenylate cyclase activation restored cell viability, thus suggesting cAMP as a principle mediator of exendin-4 anti-tumorigenic activity. While the pancreatic form of the GLP-1R could not be detected in breast cancer cells, several lines of evidence indicated the existence of an alternative GLP-1R in mammary cells. Thus, internalization of GLP-1 into MCF-7 cells was evidenced, infection of MCF-7 cells with the pancreatic receptor enhanced proliferation, and treatment with exendin-(9-39), a GLP-1R antagonist, further increased cAMP levels. Our studies indicate the incretin hormone GLP-1 as a potent inducer of cAMP and an inhibitor of breast cancer cell proliferation. Reduced GLP-1 levels may, therefore, serve as a novel link between obesity, diabetes mellitus, and breast cancer. PMID- 21638054 TI - Bone metastasis in a novel breast cancer mouse model containing human breast and human bone. AB - In practice, investigations for bone metastasis of breast cancer rely heavily on models in vivo. Lacking of such ideal model makes it difficult to study the whole process or accurate mechanism of each step of this metastatic disease. Development of xenograft mouse models has made great contributions in this area. Currently, the best animal model of breast cancer metastasizing to bone is NOD/SCID-hu models containing human bone, which makes it possible to let the breast cancer cells and the bone target of osteotropic metastasis be both of human origin. We have developed a novel mouse model containing both human bone and breast, and proved it functional and reliable. In this study, a set of human breast cancer cell line including MDA-MB-231, MDA-MB-231BO, MCF-7, ZR-75-1 and SUM1315 were characterized their osteotropism in this model. A specific cell line SUM1315 made species-specific bone metastasis, certifying the osteotropism identification utility of the novel mouse model. Furthermore, gene expression and microRNA expression profiling analysis were done to the two SUM1315 derived sub lines isolated and purified from the orthotopic and metastatic xenograft. In addition, to demonstrate the disparity between the "spontaneous" and "forced" bone metastasis in mouse model, MDA-MB-231 cells were inoculated into both the human implants in this model simultaneously, and then primary cultured and profiling analyzed. Supported by overall results of profiling analyses, this study suggested the novel model was a useful tool for understanding, preventing and treating bone metastasis of breast cancer, meanwhile it had provided significant information for further investigations. PMID- 21638055 TI - Survival of thermophilic and hyperthermophilic microorganisms after exposure to UV-C, ionizing radiation and desiccation. AB - In this study, we investigated the ability of several (hyper-) thermophilic Archaea and phylogenetically deep-branching thermophilic Bacteria to survive high fluences of monochromatic UV-C (254 nm) and high doses of ionizing radiation, respectively. Nine out of fourteen tested microorganisms showed a surprisingly high tolerance against ionizing radiation, and two species (Aquifex pyrophilus and Ignicoccus hospitalis) were even able to survive 20 kGy. Therefore, these species had a comparable survivability after exposure to ionizing radiation such as Deinococcus radiodurans. In contrast, there was nearly no difference in survival of the tested strains after exposure to UV-C under anoxic conditions. If the cells had been dried in advance of UV-C irradiation, they were more sensitive to UV-C radiation compared with cells irradiated in liquid suspension; this effect could be reversed by the addition of protective material like sulfidic ores before irradiation. By exposure to UV-C, photoproducts were formed in the DNA of irradiated Archaea and Bacteria. The distribution of the main photoproducts was species specific, but the amount of the photoproducts was only partly dependent on the applied fluence. Overall, our results show that tolerance to radiation seems to be a common phenomenon among thermophilic and hyperthermophilic microorganisms. PMID- 21638056 TI - Evolution of temperature optimum in Thermotogaceae and the prediction of trait values of uncultured organisms. AB - Quantitative characterization of the mode and rate of phenotypic evolution is rarely applied to prokaryotes. Here, we present an analysis of temperature optimum (T (opt)) evolution in the thermophilic family Thermotogaceae, which has a large number of cultured representatives. We use log-rate-interval analysis to show that T (opt) evolution in Thermotogaceae is consistent with a Brownian motion (BM) evolutionary model. The properties of the BM model are used to a establish confidence intervals on the unknown phenotypic trait value of an uncultured organism, given its distance to a close relative with known trait value. Cross-validation by bootstrapping indicates that the predictions are robust. PMID- 21638057 TI - The unresolved challenge of subjects at intermediate cardiovascular risk. PMID- 21638058 TI - The FOUR score: is it just another new coma scale? PMID- 21638059 TI - Herbal remedies in hospital setting: an Italian explorative survey among physicians and patients on knowledge and use. PMID- 21638060 TI - Unusual and voluminous pulmonary trunk aneurysm. PMID- 21638061 TI - Cloning and characterization of a maize SnRK2 protein kinase gene confers enhanced salt tolerance in transgenic Arabidopsis. AB - SnRK2 (sucrose non-fermenting 1-related protein kinases 2) represents a unique family of protein kinase in regulating signaling transduction in plants. Although the regulatory mechanisms of SnRK2 have been well demonstrated in Arabidopsis thaliana, their functions in maize are still unknown. In our study, we cloned an SnRK2 gene from maize, ZmSAPK8, which encoded a putative homolog of the rice SAPK8 protein. ZmSAPK8 had two copies in the maize genome and harbored eight introns in its coding region. We demonstrated that ZmSAPK8 expressed differentially in various organs of maize plants and was up-regulated by high salinity and drought treatment. A green fluorescent protein (GFP)-tagged ZmSAPK8 showed subcellular localization in the cell membrane, cytoplasm and nucleus. In vitro kinase assays indicated that ZmSAPK8 preferred Mn(2+) to Mg(2+) as cofactor for phosphorylation, and Ser-182 and Thr-183 in activation loop was important for its activity. Heterologous overexpression of ZmSAPK8 in Arabidopsis could significantly strengthen tolerance to salt stress. Under salt treatment, ZmSAPK8 overexpressed transgenic plants exhibited higher germination rate and proline content, low electrolyte leakage and higher survival rate than wild type. Further analysis indicated that transgenic plants showed increased transcription of the stress-related genes, RD29A, RD29B, RAB18, ABI1, DREB2A and P5CS1, under high salinity conditions. The results demonstrated that ZmSAPK8 was involved in diverse stress signal transduction. Moreover, no obvious adverse effects on growth and development in the ZmSAPK8-overexpressed transgenic plants implied that ZmSAPK8 was potentially useful in transgenic breeding to improve salt tolerance in crops. PMID- 21638062 TI - Treatment of bleached wool with trans-glutaminases to enhance tensile strength, whiteness, and alkali resistance. AB - Trans-glutaminases is known as a cross-linking enzyme for proteins. Wool is a proteinous fiber conventionally is treated through several processes to obtain the desirable characteristics. Bleaching is also one of the most important processes usually carried out by using an oxidizing agent in a conventional method. The tensile strength of wool yarns was reduced as a consequence of oxidative bleaching. Here, with the help of microbial trans-glutaminases (m TGases), a novel bleaching process was disclosed in a way to obtain a bleached wool yarn with no significant reduction in the tensile strength. The results confirmed that the bleached wool yarns with H(2)O(2) could be modified by m TGases post-treatment. The m-TGases treatment on the bleached wool yarns improved the tensile strength and whiteness along with the higher alkali resistance. PMID- 21638063 TI - Determinants of DHA incorporation into tumor tissue during dietary DHA supplementation. AB - Docosahexaenoic acid (DHA), upon incorporation into tumor tissue, has the potential to sensitize tumors to the effects of chemotherapy or radiation therapy. Although DHA has usually been supplied to tumor tissue in the diet, appropriate dietary conditions required to obtain optimal tumor levels have not been established. Hence, we studied mammary tumor tissue responses in rats fed various durations and doses of DHA. Rats fed a palm oil enriched diet (diet 0) were switched to diets providing either 0.8 g DHA/day (diet 1) or 1.5 g DHA/day (diet 2). Tumor tissue fatty acid composition was analysed at baseline (diet 0), at weeks 1, 4 and 9 during diet 1 and at week 4 during diet 2. Dietary DHA supplementation differentially increased DHA within phospholipids (PL) and triacylglycerol (TAG) fractions in tumors. DHA level equilibrated between 2 and 4 weeks in PL while DHA increase was more progressive in TAG and did not reach a steady state. A higher dose of DHA further increased DHA content in tumor PL and TAG (P = 0.018 and P < 0.001, respectively). DHA concentration in plasma PL was positively correlated with DHA in tumor PL (r = 0.72; P = 0.0003) and TAG (r = 0.64; P = 0.003). We conclude that dietary DHA supplementation enhances tumor content of DHA in a time- and dose-dependent manner, and that the DHA level in plasma PL could be used as a proxy for tumor DHA. These findings have implications for dietary DHA supplementations in cancer patients. PMID- 21638064 TI - Ethanolic extract of propolis promotes reverse cholesterol transport and the expression of ATP-binding cassette transporter A1 and G1 in mice. AB - The ethanolic extract of propolis (EEP) is beneficial in increasing high density lipoprotein (HDL) cholesterol (HDL-C) and diminishing risks of atherosclerosis. In this study, we examined the effects of EEP on reverse cholesterol transport in mice. (3)H -cholesterol laden macrophage was injected intraperitoneally into mice fed by gastric gavage with EEP. Plasma lipid level was determined and (3)H cholesterol was traced in plasma, liver and feces. The effects of EEP on ATP binding cassette transporter A1 and G1 (ABCA1 and ABCG1) and scavenger receptor BI (SR-BI) in mice liver and in cultured cells were also investigated. EEP administration led to a significant increase in HDL-C and peritoneal macrophage original (3)H-cholesterol in plasma, liver and feces. Liver protein expressions of ABCA1 and ABCG1 were increased but SR-B1 was not. In vitro experiments with HepG2 and Raw264.7 cell lines confirmed the above results. The finding of these studies shows that EEP-enhanced reverse cholesterol transport may have resulted from EEP stimulated plasma HDL level and hepatic ABCA1 and ABCG1 expression. PMID- 21638065 TI - Chlorination in a wastewater treatment plant: acute toxicity effects of the effluent and of the recipient water body. AB - This study investigates the impact of wastewater treatment plant (WWTP) effluent on the toxicity of the recipient water body and the effectiveness of the disinfection treatment applied (sodium hypochloride) to assure the compliance of both microbiological and toxicological emission limits. No toxicity was found in the majority of samples collected from the recipient river, upstream and downstream of the WWTP, using three different toxicity tests (Vibrio fischeri, Daphnia magna, and Pseudokirchneriella subcapitata). Only three samples presented toxic unit (TU) values with V. fischeri, and one presented TU with P. subcapitata. The influent toxicity ranged from slightly toxic to toxic (TU = 0.68 4.47) with V. fischeri, while only three samples presented TU values with the other tests. No toxicity was found in the absence of chlorination, while the mean toxicity was 3.42 +/- 4.12 TU with chlorination in the effluent. Although no toxicity or very slight toxicity was found in the receiving water, its residual toxicity was higher than the US EPA Quality Standard in two samples. Escherichia coli concentration had a lower mean value in the chlorinated effluent: 13,993 +/- 12,037 CFU/100 mL vs. 62,857 +/- 80,526 CFU/100 mL for the not chlorinated effluent. This difference was shown to be significant (p < 0.05). E. coli in ten chlorinated samples was higher than the limit established by European and Italian Legislation. The mean highest trihalomethanes (THMs) value was found in the influent samples (2.79 +/- 1.40 MUg/L), while the mean highest disinfection by products (DBPs) was found in the effluent samples (1.85 +/- 2.25 MUg/L). Significant correlations were found between toxicity, sodium hypochlorite, THMs, DBPs, E. coli, and residual chlorine. In conclusion, this study highlighted that the disinfection of wastewater effluents with sodium hypochlorite determines the increase of the toxicity, and sometimes is not enough to control the E. coli contamination. PMID- 21638066 TI - New evidences in the complexity of contamination of the lagoon of Venice: polybrominated diphenyl ethers (PBDEs) pollution. AB - This study presents the first evaluation of the current pollution by polybrominated diphenyl ethers (PBDEs) of surface sediments from the Lagoon of Venice. We focused the research on tri-to hepta-BDEs, the main components of penta- and octa-mixtures, which are considered to be the most toxic for the biocoenosis. The results pointed out a quite homogeneous contamination of this keystone European transitional environment, with ? (13)PBDEs values ranging from 0.39 to 6.78 ng/g dry weight; these values reflect low to moderate pollution levels, which is in conformity to other coastal European ecosystems. The average PBDE profile of the lagoon sediments follows this decreasing trend of congeners: BDE-47>BDE-99> >BDE-190>BDE-28>BDE-153>BDE-154>BDE-138, BDE-183, and BDE-17, which is similar to the worldwide distribution pattern. BDE-47 and BDE-99 revealed a recent use of a penta-BDE mixture, while the presence of hepta-BDEs (BDE-183 and BDE-190) in all of the sites can indicate the actual use of a deca BDE formulation, because these congeners are considered to be debrominated byproducts of BDE-209 degradation. PMID- 21638067 TI - EAF2 loss enhances angiogenic effects of Von Hippel-Lindau heterozygosity on the murine liver and prostate. AB - Von Hippel-Lindau (VHL) disease results from the inactivation of the VHL gene and is characterized by highly vascular tumors. A consequence of VHL loss is the stabilization of hypoxia-inducible factor (HIF) alpha subunits and increased expression of HIF target genes, which include pro-angiogenic growth factors such as vascular endothelial growth factor (VEGF). In mice, homozygous deletion of VHL is embryonic lethal due to vascular abnormalities in the placenta; and, VHL(+/-) mice develop proliferative vascular lesions in several major organs, most prominently the liver. Loss of ELL-associated factor (EAF2) in murine models has also been shown to induce increased vascular density in the liver as well as the prostate. Previously, EAF2 was determined to be a binding partner of VHL and loss of EAF2 induced a reduction in pVHL levels and an increase in hypoxia induced factor 1alpha (HIF1alpha) levels in vitro. Here we characterized the cooperative effects of VHL- and EAF2-deficiency on angiogenesis in the liver and prostate of male mice. VHL deficiency consistently increased the incidence of hepatic vascular lesions across three mouse strains. These vascular lesions where characterized by an increase in microvessel density, and staining intensity of VHL target proteins HIF1alpha and VEGF. EAF2(-/-)VHL(+/-) mice had increased incidence of proliferative hepatic vascular lesions (4/4) compared to VHL(+/-) (10/18) and EAF2(-/-) (0/5) mice. Prostates of EAF2(-/-)VHL(+/-) mice also displayed an increase in microvessel density, as well as stromal inflammation and prostatic intraepithelial neoplasia. These results suggest that cooperation of VHL and EAF2 may be critical for angiogenic regulation of the liver and prostate, and concurrent loss of these two tumor suppressors may result in a pro-angiogenic phenotype. PMID- 21638068 TI - A case of malignant hyperthermia captured by an anesthesia information management system. AB - Many cases of malignant hyperthermia triggered by volatile anesthetic agents have been described. However, to our knowledge, there has not been a report describing the precise changes in physiologic data of a human suffering from this process. Here we describe a case of malignant hyperthermia in which monitoring information was frequently and accurately captured by an anesthesia information management system. PMID- 21638069 TI - Cost of care for colorectal cancer in Ireland: a health care payer perspective. AB - OBJECTIVE: Management options for colorectal cancer have expanded in recent years. We estimated average lifetime cost of care for colorectal cancer in Ireland in 2008, from the health care payer perspective. METHOD: A decision tree model was developed in Microsoft EXCEL. Site and stage-specific treatment pathways were constructed from guidelines and validated by expert clinical opinion. Health care resource use associated with diagnosis, treatment and follow up were obtained from the National Cancer Registry Ireland (n=1,498 cancers diagnosed during 2004-2005) and three local hospital databases (n=155, 142 and 46 cases diagnosed in 2007). Unit costs for hospitalisation, procedures, laboratory tests and radiotherapy were derived from DRG costs, hospital finance departments, clinical opinion and literature review. Chemotherapy costs were estimated from local hospital protocols, pharmacy departments and clinical opinion. Uncertainty was explored using one-way and probabilistic sensitivity analysis. RESULTS: In 2008, the average (stage weighted) lifetime cost of managing a case of colorectal cancer was ?39,607. Average costs were 16% higher for rectal (?43,502) than colon cancer (?37,417). Stage I disease was the least costly (?23,688) and stage III most costly (?48,835). Diagnostic work-up and follow-up investigations accounted for 4 and 5% of total costs, respectively. Cost estimates were most sensitive to recurrence rates and prescribing of biological agents. CONCLUSION: This study demonstrates the value of using existing data from national and local databases in contributing to estimating the cost of managing cancer. The findings illustrate the impact of biological agents on costs of cancer care and the potential of strategies promoting earlier diagnosis to reduce health care resource utilisation and care costs. PMID- 21638070 TI - Strength training improves cycling efficiency in master endurance athletes. AB - The purpose of this study was to test the effect of a 3-week strength training program of knee extensor muscles on cycling delta efficiency in master endurance athletes. Nine master (age 51.5 +/- 5.5 years) and 8 young (age 25.6 +/- 5.9 years) endurance athletes with similar training levels participated in this study. During three consecutive weeks, all the subjects were engaged in a strength training program of the knee extensor muscles. Every week, they performed three training sessions consist of 10 * 10 knee extensions at 70% of maximal repetition with 3 min rest between in a leg extension apparatus. Maximal voluntary contraction torque (MVC torque) and force endurance (End) were assessed before, after every completed week of training, and after the program. Delta efficiency (DE) in cycling was evaluated before and after the training period. Before the training period, MVC torque, End, and DE in cycling were significantly lower in masters than in young. The strength training induced a significant improvement in MVC torque in all the subjects, more pronounced in masters (+17.8% in masters vs. +5.9% in young, P < 0.05). DE in cycling also significantly increased after training in masters, whereas it was only a trend in young. A significant correlation (r = 0.79, P < 0.01) was observed between MVC torque and DE in cycling in masters. The addition of a strength training program for the knee extensor muscles to endurance-only training induced a significant improvement in strength and cycling efficiency in master athletes. This enhancement in muscle performance alleviated all the age-related differences in strength and efficiency. PMID- 21638071 TI - Are tangles as toxic as they look? AB - Neurofibrillary tangles are intracellular accumulations of hyperphosphorylated and misfolded tau protein characteristic of Alzheimer's disease and other tauopathies. Classic cross-sectional studies of Alzheimer patient brains showed associations of tangle accumulation with neuronal loss, synapse loss, and dementia, which led to the supposition that tangles are toxic to neurons. More recent advances in imaging techniques and mouse models have allowed the direct exploration of the question of toxicity of aggregated versus soluble tau and have surprisingly challenged the view of tangles as toxic species in the brain. Here, we review these recent experiments on the nature of the toxicity of tau with particular emphasis on our experiments imaging tangles in the intact brain through a cranial window, which allows observation of tangle formation and longitudinal imaging of the fate of tangle-bearing neurons. PMID- 21638072 TI - Pseudohypoaldosteronism masquerading as congenital adrenal hyperplasia. AB - A 15-day- old male infant presented with features suggestive of congenital adrenal hyperplasia (CAH). But on detailed investigation, he had normal 17-OH Progesterone and high aldosterone levels. Infant also had right sided hydronephrosis and associated urinary tract infection. Hence, a diagnosis of secondary pseudohypoaldosteronism was made. There was good response to sodium chloride supplementation and he was thriving well on follow up. While evaluating any infant with suspected CAH, one should consider secondary pseudohypoaldosteronism, especially when risk factors like urinary tract obstruction or UTI coexists. PMID- 21638073 TI - Evaluation of health related quality of life in 6-18 years old patients with acute leukemia during chemotherapy. AB - OBJECTIVE: To evaluate the quality of life (QOL) of Iranian children with acute leukemia during chemotherapy. METHODS: One hundred patients between 6 to 18-years old were selected by convenient sampling method. EORTC QLQ-C30 Questionnaire was completed by their parents' help. Demographic information such as age, sex and type of leukemia were also collected. These data were evaluated by SPSS software, Chi-square and independent sample T test. The relation between different scales of questionnaire and variables was measured and final results were compared with reference values. RESULTS: In acute Lymphoblastic Leukemia patients QOL, physical and cognitive functions were lower in comparison with acute myelogenous leukemia and they had more fatigue, pain and insomnia. The patients between ages of 12-18 years-old had more financial difficulties and diarrhea and lower cognitive function in comparison with 6-12-years-old patients. The present patients achieved higher scores than reference value, but they had more economic problem. CONCLUSIONS: The patients had relatively good QOL. The lowest impression was in cognitive function and the highest was in emotional function. The patients mostly complained of financial difficulties and fatigue and rarely of diarrhea and constipation. It is necessary to do more researches related to health related QOL in pediatric patients. PMID- 21638074 TI - Effect of pseudoephedrine on cardiac rhythm of children with rhinitis. AB - OBJECTIVE: To investigate the effect of pseudoephedrine on heart rhythm of children with rhinitis. METHODS: The study included 25 children diagnosed with rhinitis from March 2009 through February 2010 in the Department of Pediatrics. Holter records were obtained for 24 h before and at the fourth day of pseudoephedrine treatments. RESULTS: Study group consisted of 18 girls (72%) and 7 boys (28%) with a mean age of 8.7 +/- 3.4 (4-17.9 years). Common complaints of the patients were rhinorrhea (100%), cough (68%) fatigue (48%), sore throat (36%), and headache (28%). Of the 25 patients whose Holter recordings were evaluated, rare supraventricular extrasystoles were observed in one prior to the administration of pseudoephedrine, which were not repeated on this patient's follow-up recording on day four. There were two ventricular extrasystoles in the day four Holter recording of another patient. None of the patients complained of chest pain or palpitation. There were no observations of supraventricular tachycardia, ventricular tachycardia or ventricular fibrillation. No statistical differences could be found (p > 0.05) in the values before treatment and those on day four of treatment of either the time-dependent Heart rate variability (HRV) parameters SDNN, SDNN index, SDANN and RMSSD, or the frequency-dependent parameters (TP, HF, LF). No statistical difference could be determined between heart rate values of the patients before treatment and those on day four of treatment (p > 0.05). CONCLUSIONS: This study has established that therapeutic doses of pseudoephedrine do not cause an additional dysrhythmia risk for children with no health problem except rhinitis. PMID- 21638075 TI - An endoscopic modification of the simultaneous 'above and below' approach to large pituitary adenomas. AB - Surgical resections of large-to-giant pituitary adenomas (PA) are technically challenging procedures. Tumors with a fibrous consistency or 'hour-glass' configurations are particularly difficult to remove completely and safely through the transsphenoidal route alone. Although the transcranial approach can facilitate the removal of a large suprasellar mass, it may be associated with significant bleeding within the intradural space. A simultaneous microscopic transcranial and transsphenoidal approach has been described as an alternative surgical strategy. We have further modified this 'above and below' approach by adopting endoscopic techniques for the transsphenoidal part of the procedure. This modified approach has the advantages of requiring only one operating microscope, and permitting freer maneuvers and easier orientation for both surgical teams. We present two patients successfully treated with this approach. Complete tumor removal was achieved and both patients achieved satisfactory functional recovery. PMID- 21638076 TI - Silver discharged in effluents from image-processing services: a risk to human and environmental health. AB - In Brazil, only 20.2% of the municipalities have sewage collection and treatment. The use of medical diagnostic by radiographic processing generates effluents which may contain contaminants such as silver (Ag) that causes irreversible damage to health. They can also contaminate soil, water, and food if there is no treatment before disposal. This study aimed to identify and quantify the Ag concentration in radiographic film-washing water and fixer generated from radiographic processing in 12 Brazilian health institutions, in order to offer subsidies to the environmental and public health managers about this issue. The Ag values found in the washing water films samples ranged from 0.1 to 1,785.97 mg L(-1). In the fixer samples, the values ranged from 435.59 to 16,325.92 mg L(-1). These data are far above the values required by Brazilian legislation whose limit is 0.1 mg L(-1) for Ag in effluents released directly or indirectly into the environment. This research reveals that the disposal of Ag in this kind of effluent needs to be monitored and controlled by the Brazilian government in order to protect both the human and environment health from those contaminants. PMID- 21638077 TI - Impairment of breast cancer cell invasion by COX-2-specific inhibitor NS398: roles of CXCR4 and of uPA system. AB - Inhibition of cyclooxygenase-2 (COX-2) is known to impair cancer cell metastatic behaviour, but the mechanisms involved largely remain elusive. We aimed to analyse whether the antimetastatic effect of COX-2 inhibition in breast cancer cells could be explained by variations in the expression levels of chemokine receptor CXCR4, vascular endothelium growth factor (VEGF) and UPA/UPAR components of the urokinase plasminogen activator system (uPAR). Breast cancer cell line MDA MB-231 was exposed to COX-2-specific inhibitor NS398. Experimental data were assessed using Matrigel invasion tests, qRT-PCR, ELISA, flow cytometry and MTT test. Exposure to NS398 had no major effect on cell viability, apoptosis or VEGF production. Cell invasion was significantly decreased with reductions ranging from of 3.6% with 10 MUM NS398 to 81.04% with 100 MUM NS398. CXCR4 membrane expression was significantly reduced by 18% (P < 0.05) when cells were treated with 100 MUM of NS398 for 72 h. UPA mRNA levels were significantly reduced to 78 and 63% after treatment with 10 MUM NS398 for 48 and 72 h, respectively (P < 0.05). UPAR mRNA levels also decreased with mild NS398 concentrations, reaching the lowest level of 56% with 50 MUM of NS398 for 48 h (P < 0.05). With NS398 higher concentrations, UPAR and UPA expression levels increased. According to our results, impairment of expression of CXCR4, UPA and UPAR differentially contribute to the antimetastatic effect of COX-2 inhibitors depending on drug concentration. PMID- 21638078 TI - Role of urokinase plasminogen activator receptor (CD87) as a prognostic marker in acute myeloid leukemia. AB - Expression of uPAR (CD87) may play a relevant role in the diagnosis and pathophysiology and prognostic pattern of acute myeloid leukemia. The aims of this study were to find out the prognostic significance of pretreatment detection of CD87 and study the prevalence of CD87 expression and its value as a predictor for survival in AML patients. CD87 expression has been studied on blasts in 110 newly diagnosed AML patients. Immunophenotyping and cytogenetic analysis of these cases were performed. CD87 was positive in 80 (72.7%) cases of patients. The MFI categorized the expression of CD87 into CD87(High) and CD87(Low) expression. Blast cells show that monocytic differentiation has a significantly more CD87(High) expression than CD87(Low) expression. Cytogenetic abnormalities were found in 58.7% of patients with CD87(Low) AML and 41.25% of patients with CD87(High) AML. Cases with CD87(High) expression cells were characterized by a significantly lower survival period especially when co-expressed with CD56, CD34, and/or CD64. There is a negative prognostic influence of the expression of CD87 on the surface of AML blasts, but more tests are necessary to explain the pathophysiological mechanisms behind these findings and to learn about the mechanism that influences the CD87 expression and function. PMID- 21638079 TI - The vulnerability cube: a multi-dimensional framework for assessing relative vulnerability. AB - The diversity and abundance of information available for vulnerability assessments can present a challenge to decision-makers. Here we propose a framework to aggregate and present socioeconomic and environmental data in a visual vulnerability assessment that will help prioritize management options for communities vulnerable to environmental change. Socioeconomic and environmental data are aggregated into distinct categorical indices across three dimensions and arranged in a cube, so that individual communities can be plotted in a three dimensional space to assess the type and relative magnitude of the communities' vulnerabilities based on their position in the cube. We present an example assessment using a subset of the USEPA National Estuary Program (NEP) estuaries: coastal communities vulnerable to the effects of environmental change on ecosystem health and water quality. Using three categorical indices created from a pool of publicly available data (socioeconomic index, land use index, estuary condition index), the estuaries were ranked based on their normalized averaged scores and then plotted along the three axes to form a vulnerability cube. The position of each community within the three-dimensional space communicates both the types of vulnerability endemic to each estuary and allows for the clustering of estuaries with like-vulnerabilities to be classified into typologies. The typologies highlight specific vulnerability descriptions that may be helpful in creating specific management strategies. The data used to create the categorical indices are flexible depending on the goals of the decision makers, as different data should be chosen based on availability or importance to the system. Therefore, the analysis can be tailored to specific types of communities, allowing a data rich process to inform decision-making. PMID- 21638081 TI - Aligning patient preferences and patient care at the end of life. PMID- 21638082 TI - The role of PERK and GCN2 in basal and hydrogen peroxide-regulated translation from the hepatitis C virus internal ribosome entry site. AB - We have previously shown that translation from the HCV IRES is up-regulated by patho/physiological doses of H(2)O(2) but is still sensitive to the inhibitory effect of phospho-eIF2alpha in hepatocytes. In this study using wild type and 'knockout' mouse embryonic fibroblasts (MEFs), we showed that two of the eIF2alpha kinases, PERK and GCN2, were not responsible for translational regulation under physiological and a higher apoptotic doses of H(2)O(2) (100 MUM). However, a differential translational response was observed at a lower apoptotic dose of H(2)O(2) (50 MUM) between Perk+/+ and Perk-/- MEFs but not that between Gcn2+/+ and Gcn2-/- MEFs, suggesting that PERK may play a role in translational up-regulation under oxidative stress. Our results also suggest that PERK mediates such an effect via an eIF2-independent pathway. This is in contrast to the canonical role of PERK on translational inhibition under stress conditions via phosphorylation of eIF2alpha. When tested for the role of PERK and GCN2 on basal translation from the HCV IRES under non-stressed condition, we found that basal translation from the HCV IRES was also favoured in the presence of PERK or GCN2 in MEFs over that of cap-dependent translation and was favoured in the presence of GCN2 but not PERK in Huh-7 cells. These results suggest that PERK and GCN2 also have a functional role on regulating translation under non-stressed conditions, apart from their long established roles as stress kinases. PMID- 21638083 TI - Beneficial effects of theophylline infusions in surgical patients with intra abdominal hypertension. AB - BACKGROUND: Intra-abdominal hypertension (IAH) can cause high mortality. Recently, we found that IAH was associated with increased serum levels of adenosine and interleukin 10. Our present "hypothesis-generated study" was based on the above mentioned results. MATERIALS AND METHODS: In this uncontrolled clinical trial, a total of 78 patients with IAH were enrolled representing a 13 20 mmHg range of intra-abdominal pressure (IAP). Patients requiring surgical abdominal decompression were excluded. Patients were treated with the following protocols: standard supportive therapy (ST, n = 38) or ST plus infusion with the adenosine receptor antagonist theophylline (T, n = 40). Over the 5-day measurement period, IAP was monitored continuously and serum adenosine concentration and other clinical and laboratory measurements were monitored daily. Mortality was followed for the first 30 days following the diagnosis of IAH. RESULTS: Mortality of ST patients was 55%, which is compatible to other studies. Serum adenosine concentration was found to be directly proportional to IAP. Of the 40 patients receiving T treatment, survival was 100%. An increased survival related to theophylline infusion correlated with improving serum concentrations of IL-10, urea, and creatinine, as well as 24-h urine output, fluid balance, mean arterial pressure, and O(2)Sat. CONCLUSIONS: Adenosine receptor antagonism with T following IAH diagnosis resulted in markedly reduced mortality in patients with moderated IAH (<20 mmHg). Theophylline-associated mortality reduction may be related to improved renal perfusion and improved MAP, presumably caused by adenosine receptor blockade. Because this study was not a randomized controlled study, these compelling observations require further multicentric clinical confirmation. PMID- 21638084 TI - RGD-conjugated human ferritin nanoparticles for imaging vascular inflammation and angiogenesis in experimental carotid and aortic disease. AB - PURPOSE: Inflammation and angiogenesis are important contributors to vascular disease. We evaluated imaging both of these biological processes, using Arg-Gly Asp (RGD)-conjugated human ferritin nanoparticles (HFn), in experimental carotid and abdominal aortic aneurysm (AAA) disease. PROCEDURES: Macrophage-rich carotid lesions were induced by ligation in hyperlipidemic and diabetic FVB mice (n = 16). AAAs were induced by angiotensin II infusion in apoE(-/-) mice (n=10). HFn, with or without RGD peptide, was labeled with Cy5.5 and injected intravenously for near-infrared fluorescence imaging. RESULTS: RGD-HFn showed significantly higher signal than HFn in diseased carotids and AAAs relative to non-diseased regions, both in situ (carotid: 1.88 +/- 0.30 vs. 1.17 +/- 0.10, p = 0.04; AAA: 2.59 +/- 0.24 vs. 1.82 +/- 0.16, p = 0.03) and ex vivo. Histology showed RGD-HFn colocalized with macrophages in carotids and both macrophages and neoangiogenesis in AAA lesions. CONCLUSIONS: RGD-HFn enhances vascular molecular imaging by targeting both vascular inflammation and angiogenesis, and allows more comprehensive detection of high-risk atherosclerotic and aneurysmal vascular diseases. PMID- 21638085 TI - Towards a sociology of knowledge translation: the importance of being dis interested in knowledge translation. AB - Over the last 15 years there has been an increasingly energetic search for theories and definitions in the burgeoning area of knowledge translation (KT) in the health care context. The focus has been on the design and evaluation of KT activities with little attention to developing a considered KT theoretical/methodological approach that takes a more distanced critical inquiry to the studying of KT interventions. As such, what has been overlooked in the health professions KT literature to date is a suitably complex conceptualisation of translation that encompasses the multiplicity of KT interventions, the dynamic environments in which they are occurring, and the production of new knowledge which arises from their interactions. Attending to these epistemological issues; i.e. issues of the type of knowledge and how it is produced, is crucial to developing a relational understanding of the production and emergence of context specific clinical knowledge and practice in KT processes. Such an approach is compared to the traditional KT evaluations by KT practitioners themselves of the compliance or non-compliance of individual and organisational behaviour change. A Sociology of KT (SKT) is proposed to complement the applied and evaluative research practice of implementing planned action KT activities. The purpose of a SKT basic research agenda is to understand both the context-specific nature of KT interventions and the epistemological premises of the activities of KT implementers themselves, in order to advance the science of KT and inform, complement and add to the success of applied and evaluative KT research in the future. PMID- 21638086 TI - Core components of communication of clinical reasoning: a qualitative study with experienced Australian physiotherapists. AB - Communication is an important area in health professional education curricula, however it has been dealt with as discrete skills that can be learned and taught separate to the underlying thinking. Communication of clinical reasoning is a phenomenon that has largely been ignored in the literature. This research sought to examine how experienced physiotherapists communicate their clinical reasoning and to identify the core processes of this communication. A hermeneutic phenomenological research study was conducted using multiple methods of text construction including repeated semi-structured interviews, observation and written exercises. Hermeneutic analysis of texts involved iterative reading and interpretation of texts with the development of themes and sub-themes. Communication of clinical reasoning was perceived to be complex, dynamic and largely automatic. A key finding was that articulating reasoning (particularly during research) does not completely represent actual reasoning processes but represents a (re)construction of the more complex, rapid and multi-layered processes that operate in practice. These communications are constructed in ways that are perceived as being most relevant to the audience, context and purpose of the communication. Five core components of communicating clinical reasoning were identified: active listening, framing and presenting the message, matching the co communicator, metacognitive aspects of communication and clinical reasoning abilities. We propose that communication of clinical reasoning is both an inherent part of reasoning as well as an essential and complementary skill based on the contextual demands of the task and situation. In this way clinical reasoning and its communication are intertwined, providing evidence for the argument that they should be learned (and explicitly taught) in synergy and in context. PMID- 21638087 TI - The cerebellar component of Friedreich's ataxia. AB - Lack of frataxin in Friedreich's ataxia (FRDA) causes a complex neurological and pathological phenotype. Progressive atrophy of the dentate nucleus (DN) is a major intrinsic central nervous system lesion. Antibodies to neuron-specific enolase (NSE), calbindin, glutamic acid decarboxylase (GAD), and vesicular glutamate transporters 1 and 2 (VGluT1, VGluT2) allowed insight into the disturbed synaptic circuitry of the DN. The available case material included autopsy specimens of 24 patients with genetically defined FRDA and 14 normal controls. In FRDA, the cerebellar cortex revealed intact Purkinje cell somata and dendrites as assessed by calbindin immunoreactivity. The DN, however, displayed severe loss of large NSE-reactive neurons. Small neurons remained intact. Labeling of Purkinje cells, basket fibers, Golgi neurons, and Golgi axonal plexuses with antibodies to GAD indicated normal intrinsic circuitry of the cerebellar cortex involving gamma-aminobutyric acid (GABA). In contrast, the DN displayed severe loss of GABA-ergic terminals and formation of GAD- and calbindin reactive grumose degeneration. The surviving small GAD-positive DN neurons provided normal GABA-ergic terminals to intact inferior olivary nuclei. The olives also received normal glutamatergic terminals as shown by VGluT2 reactivity. VGluT1-immunocytochemistry of the cerebellar cortex confirmed normal glutamatergic input to the molecular layer by parallel fibers and the granular layer by mossy fibers. VGluT2-immunoreactivity visualized normal climbing fibers and mossy fiber terminals. The DN, however, showed depletion of VGluT1- and VGluT2-reactive terminals arising from climbing and mossy fiber collaterals. The main functional deficit underlying cerebellar ataxia in FRDA is defective processing of inhibitory and excitatory impulses that converge on the large neurons of the DN. The reason for the selective vulnerability of these nerve cells remains elusive. PMID- 21638088 TI - Assessment of BRAF V600E mutation status by immunohistochemistry with a mutation specific monoclonal antibody. AB - Activating mutations of the serine threonine kinase v-RAF murine sarcoma viral oncogene homolog B1 (BRAF) are frequent in benign and malignant human tumors and are emerging as an important biomarker. Over 95% of BRAF mutations are of the V600E type and specific small molecular inhibitors are currently under pre clinical or clinical investigation. BRAF mutation status is determined by DNA based methods, most commonly by sequencing. Here we describe the development of a monoclonal BRAF V600E mutation-specific antibody that can differentiate BRAF V600E and wild type protein in routinely processed formalin-fixed and paraffin embedded tissue. A total of 47 intracerebral melanoma metastases and 21 primary papillary thyroid carcinomas were evaluated by direct sequencing of BRAF and by immunohistochemistry using the BRAF V600E mutation-specific antibody clone VE1. Correlation of VE1 immunohistochemistry and BRAF sequencing revealed a perfect match for both papillary thyroid carcinomas and melanoma metastases. The staining intensity in BRAF V600E mutated tumor samples ranged from weak to strong. The generally homogenous VE1 staining patterns argue against a clonal heterogeneity of the tumors investigated. Caution is essential when only poorly preserved tissue is available for VE1 immunohistochemical analysis or when tissues with only little total BRAF protein are analyzed. Immunohistochemistry using antibody VE1 may substantially facilitate molecular analysis of BRAF V600E status for diagnostic, prognostic, and predictive purposes. PMID- 21638089 TI - Caval clamping during total hepatectomy with caval preservation in liver transplantation. PMID- 21638091 TI - Clinical scoring systems for stratifying risk after resection of hepatic colorectal metastases: still relevant? PMID- 21638090 TI - Development of the PedsQLTM Sickle Cell Disease Module items: qualitative methods. AB - PURPOSE: The objective of this qualitative study was to develop the items and support the content validity of the PedsQLTM Sickle Cell Disease Module for pediatric patients with sickle cell disease (SCD). METHODS: The iterative process included multiphase qualitative methodology. A literature review on SCD was conducted to generate domains of interest for the individual in-depth interviews. Ten healthcare experts with clinical experience in SCD participated in the development of the conceptual framework. A total of 13 pediatric patients with SCD ages 5-18 and 18 parents of patients ages 2-18 participated in the individual in-depth interviews. A total of 33 pediatric patients with SCD ages 5-18 and 39 parents of patients ages 2-18 participated in individually conducted cognitive interviews that included both think aloud and cognitive debriefing techniques to assess the interpretability and readability of the item stems. RESULTS: Six domains were derived from the qualitative methods involving patient/parent interviews and expert opinion, with content saturation achieved, resulting in 48 items. The six domains consisted of items measuring Pain Intensity/Location (9 items), Pain Interference (11 items), Worry (7 items), Emotions (3 items), Disease Symptoms/Treatment, (12 items), and Communication (6 items). CONCLUSIONS: Qualitative methods involving pediatric patients and parents in the item development process support the content validity for the PedsQLTM SCD Module. The PedsQLTM SCD Module is now undergoing national multisite field testing for the psychometric validation phase of instrument development. PMID- 21638092 TI - HIPEC for peritoneal carcinomatosis: does an associated urologic procedure increase morbidity? AB - PURPOSE: To report the incidence of urinary tract procedures performed during complete cytoreductive surgery (CCRS) plus intraperitoneal chemotherapy, and to report the types of procedure, specific morbidity, risk factors, and treatment. METHODS: Data were extracted from a prospective database of patients with malignant peritoneal disease treated with CCRS plus intraperitoneal chemotherapy who had undergone a resection or suture of the bladder, ureter, or kidney. Patients were eligible whatever the tumor origin. RESULTS: Between 1994 and 2010, among the 598 patients treated with CCRS plus intraperitoneal chemotherapy, 48 (8%) had undergone a resection or suture in the urinary tract. Procedures included 4 nephrectomies, 19 partial cystectomies, 8 surgically repaired bladder injuries, and 18 ureteral resections. Postoperative mortality was 4% and morbidity was 41%. Specific complications included 6 urinary fistulas (12%), two among the 27 bladder sutures (7%) and four among the 18 ureteral sutures (22%) (P = NS). In the multivariate analysis, the risk factors for urinary fistula were severe preoperative malnutrition (P = 0.05, relative risk [RR] = 7.3) and extensive peritoneal disease (peritoneal cancer index >=20, P = 0.05, RR = 8.3). Urinary fistulas had been treated nonsurgically in most of the cases. CONCLUSIONS: Associated urinary tract procedures had occurred in 8% of the cases but did not greatly increase morbidity. Therefore, urinary tract involvement or injury are not contraindications to performing CCRS plus intraperitoneal chemotherapy. Fistulas had complicated only 12% of urinary sutures, mainly in cases of malnutrition or extensive peritoneal disease. PMID- 21638093 TI - Is the clinical risk score for patients with colorectal liver metastases still useable in the era of effective neoadjuvant chemotherapy? AB - BACKGROUND: Several clinical risk scores (CRSs) for the outcome of patients with colorectal liver metastases have been validated, but not in patients undergoing neoadjuvant chemotherapy. Therefore, this study evaluates the predictive value of these CRSs in this specific group. METHODS: Between January 2000 and December 2008, all patients undergoing a metastasectomy were analyzed and divided into two groups: 193 patients did not receive neoadjuvant chemotherapy (group A), and 159 patients received neoadjuvant chemotherapy (group B). In group B, the CRSs were calculated before and after administration of neoadjuvant chemotherapy. Results were evaluated by using the CRSs proposed by Nordlinger et al., Fong et al., Nagashima et al., and Konopke et al. RESULTS: In groups A and B, the overall median survival was 43 and 47 months, respectively (P = 0.648). In group A, all CRSs used were of statistically significant predictive value. Before administration of neoadjuvant chemotherapy, only the Nordlinger score was of predictive value. After administration of neoadjuvant chemotherapy, all CRSs were of predictive value again, except for the Konopke score. CONCLUSIONS: Traditional CRSs are not a reliable prognostic tool when used in patients before treatment with neoadjuvant chemotherapy. However, CRSs assessed after the administration of neoadjuvant chemotherapy are useful to predict prognosis. PMID- 21638094 TI - The impact of calculation procedure on the hazard ratio of N classifications. PMID- 21638095 TI - Usefulness of the neutrophil-to-lymphocyte ratio in predicting short- and long term mortality in breast cancer patients. AB - BACKGROUND: The neutrophil-to-lymphocyte ratio (NLR) is a strong predictor of mortality in patients with colorectal, gastric, hepatocellular, pancreatic, and lung cancer. To date, the utility of NLR to predict mortality in breast cancer patients has not been studied. Therefore, the aim of our study was to determine whether the NLR is predictive of short- and long-term mortality in breast cancer patients. METHODS: Our observational study used an unselected cohort of breast cancer patients treated at the Staten Island University Hospital between January 2004 and December 2006. A total of 316 patients had a differential leukocyte count recorded prior to chemotherapy. Survival status was retrieved from our cancer registry and Social Security death index. Survival analysis, stratified by NLR quartiles, was used to evaluate the predictive value of NLR. RESULTS: Patients in the highest NLR quartile (NLR > 3.3) had higher 1-year (16% vs 0%) and 5-year (44% vs 13%) mortality rates compared with those in the lowest quartile (NLR < 1.8) (P < .0001). Those in the highest NLR quartile were statistically significantly older and had more advanced stages of cancer. After adjusting for the factors affecting the mortality and/or NLR (using two multivariate models), NLR level > 3.3 remained an independent significant predictor of mortality in both models (hazard ratio 3.13, P = .01) (hazard ratio 4.09, P = .002). CONCLUSION: NLR is an independent predictor of short- and long term mortality in breast cancer patients with NLR > 3.3. We suggest prospective studies to evaluate the NLR as a simple prognostic test for breast cancer. PMID- 21638096 TI - The diagnostic values of ultrasound and ultrasound-guided fine needle aspiration in subcentimeter-sized thyroid nodules. AB - BACKGROUND: The diagnostic performances of ultrasound (US) examination and US guided fine needle aspiration (US-FNA) were investigated in thyroid nodules <=10 mm in size. METHODS: From April 2006 to December 2006, 1440 nodules <=10 mm in size in 1403 patients (mean age, 49.3 years; range, 10-84 years) underwent US and US-FNA. The association between nodule size and inadequate specimen was investigated using the Cochran-Armitage trend test and multivariate logistic regression analysis. To evaluate the diagnostic performances of US and US-FNA, we selected 852 nodules that had undergone surgery, follow-up US-FNA, or follow-up US. A receiver operating characteristic (ROC) curve analysis was used to determine the accuracies of US and US-FNA. RESULTS: Of 1440 nodules, 256 (17.8%) yielded inadequate specimens. As the nodule size increased, the rate of inadequate specimens decreased (P < 0.001). A size of 6 mm demonstrated statistical significance between inadequate and adequate specimens (P < 0.001). The diagnostic accuracy of US was slightly improved as nodule size increased. The false positive rate of US examination was higher in nodules <=6 mm compared with that of nodules >6 mm in size (P = 0.049). However, US-FNA demonstrated high diagnostic performance in all nodules with adequate specimen. CONCLUSIONS: The inadequate specimens of US-FNA and false positives for US examinations increased as nodule size decreased. However, US-FNA demonstrated good diagnostic accuracy, assuming the specimen was adequate. PMID- 21638097 TI - Different expression patterns of CEACAM1 and its impacts on angiogenesis in gastric nonneoplastic and neoplastic lesions. AB - OBJECTIVE: This study was designed to investigate the expression patterns of CEACAM1 and its relationship with angiogenesis in nonneoplastic and neoplastic gastric lesions. METHODS: CEACAM1 and TGF-beta expression was detected by immunohistochemical staining and dual-labeling immunohistochemical staining in neoplastic and nonneoplastic lesions. MVD-CD31 and MVD-CD105 were counted in CEACAM1-positive areas by dual-labeling immunohistochemistry. RESULTS: There was no expression of CEACAM1 in normal gastric mucosa. In IM and GIN, CEACAM1 was mainly expressed with membranous pattern. CEACAM1 was expressed with membranous pattern in well-differentiated adenocarcinoma, with cytoplasmic pattern in poorly differentiated adenocarcinoma, and with cytoplasmic and membranous pattern mixed together in intermediately adenocarcinoma. The expression patterns of CEACAM1 showed a significant difference (P < 0.05) in nonneoplastic and neoplastic lesions. Coexpression of CEACAM1 and TGF-beta was elevated and significantly different from nonneoplastic to neoplastic lesions (P < 0.05). Moreover, CEACAM1 and TGF-beta coexpression were related to carcinoma progression (r = 0.35; P < 0.05). MVD-CD31 and MVD-CD105 showed significant differences from nonneoplastic to neoplastic lesions (P < 0.05). CONCLUSIONS: CEACAM1 has different expression patterns in nonneoplastic and neoplastic lesions. The coexpression of CEACAM1 and TGF-beta increased from nonneoplastic to neoplastic lesions and may be related with tumor progression via promoting tumorous angiogenesis. PMID- 21638098 TI - Analysis of margin index as a method for predicting residual disease after breast conserving surgery in a European cancer center. AB - INTRODUCTION: Breast-conserving surgery (BCS), followed by appropriate adjuvant therapies is established as a standard treatment option for women with early stage invasive breast cancers. A number of factors have been shown to correlate with local and regional disease recurrence. Although margin status is a strong predictor of disease recurrence, consensus is yet to be established on the optimum margin necessary. Margenthaler et al. recently proposed the use of a "margin index," combining tumor size and margin status as a predictor of residual disease after BCS. We applied this new predictive tool to a population of patients with primary breast cancer who presented to a symptomatic breast unit to determine its suitability in predicting those who require reexcision surgery. METHODS: Retrospective analysis of our breast cancer database from January 1, 2000 to June 30, 2010 was performed, including all patients who underwent BCS. Of 531 patients who underwent BCS, 27.1% (144/531) required further reexcision procedures, and 55 were eligible for inclusion in the study. Margin index was calculated as: margin index = closest margin (mm)/tumor size (mm) * 100, with index >5 considered optimum. RESULTS: Of the 55 patients included, 31% (17/55) had residual disease. Fisher's exact test showed margin index not to be a significant predictor of residual disease on reexcision specimen (P = 0.57). Of note, a significantly higher proportion of our patients presented with T2/3 tumors (60% vs. 38%). CONCLUSIONS: Although an apparently elegant tool for predicting residual disease after BCS, we have shown that it is not applicable to a symptomatic breast unit in Ireland. PMID- 21638099 TI - Sarcoma lung metastases treated with percutaneous radiofrequency ablation: results from 29 patients. AB - PURPOSE: We aimed to determine safety and efficacy of radiofrequency ablation (RFA) in the treatment of lung metastases arising from sarcoma. METHODS: Between 2002 and 2009, 29 patients (mean age 51 years) treated for metastatic sarcoma with a maximum of 5 lung metastases treatable with RFA were followed prospectively. The end points were local efficacy (assessed by computed tomography during the follow-up period), complications, and survival (overall and disease-free). RESULTS: A total of 47 metastases were treated with RFA. Median follow-up time was 50 months (range 28-72 months). Pneumothorax was the most frequent complication and occurred in 68.7% of the procedures. The 1- and 3-year survival rates were 92.2% (95% confidence interval [CI] 0.73-0.98) and 65.2% (95% CI 0.42-0.81), respectively. Disease-free survival was 7 months (95% CI 3.5-10). Five recurrences on RFA sites were noted during follow-up. CONCLUSIONS: RFA is safe and efficient in the treatment of lung metastasis originating from sarcomas. RFA may provide a low-morbidity alternative to surgery, being less invasive and preserving the patient's ability to undergo possible repeat operations. PMID- 21638100 TI - Amphotericin B delivery from bone cement increases with porosity but strength decreases. AB - BACKGROUND: Amphotericin B is a highly hydrophobic antifungal used for orthopaedic infections. There is disagreement about whether amphotericin B is released when it is loaded in polymethylmethacrylate (PMMA). It is unknown how much a poragen will increase amphotericin B release or decrease the compressive strength of the PMMA. QUESTIONS/PURPOSES: We therefore measured amphotericin B release and the compressive strength of amphotericin B loaded bone cement with and without adding high-dose poragen. METHODS: Antifungal-loaded bone cement was formulated with Simplex P cement and 200 mg amphotericin B with and without 10 g cefazolin (poragen) per batch. Twenty standardized test cylinders were eluted in deionized water for each formulation. Cumulative amphotericin B mass and compressive strength were measured. Data were analyzed using repeated-measures analysis of variance. RESULTS: Antifungal-loaded bone cement (ALBC) with 10 g poragen delivered more amphotericin B than ALBC containing amphotericin B alone by Day 15, 12.76 MUg/cylinder (0.5%) versus 1.74 MUg/cylinder (0.04%), respectively. With amphotericin B alone, compressive strength was unchanged and compressive strength did not decrease during elution. Adding 10 g poragen to ALBC with 200 mg amphotericin B decreased the compressive strength and compressive strength decreased further during elution, 80, 61, and 46 MPa at 0, 1, and 30 days, respectively. CONCLUSIONS: Amphotericin B is released in very small amounts from antifungal-loaded bone cement. Release can be increased by adding high-dose poragen, but compressive strength decreases sufficiently to limit its use for implant fixation. PMID- 21638102 TI - Role of ACE2 in diastolic and systolic heart failure. AB - A novel angiotensin-converting enzyme (ACE) homolog, named ACE2, is a monocarboxypeptidase which metabolizes several peptides. ACE2 degrades Angiotensin (Ang) II, a peptide with vasoconstrictive/proliferative effects, to generate Ang-(1-7), which acting through its receptor Mas exerts vasodilatory/anti-proliferative actions. In addition, as ACE2 is a multifunctional enzyme and its actions on other vasoactive peptides can also contribute to its vasoactive effects including the apelin-13 and apelin-17 peptides. The discovery of ACE2 corroborates the establishment of two counter regulatory arms within the renin-angiotensin system. The first one is formed by the classical pathway involving the ACE-Ang II-AT(1) receptor axis and the second arm is constituted by the ACE2-Ang 1-7/Mas receptor axis. Loss of ACE2 enhances the adverse pathological remodeling susceptibility to pressure-overload and myocardial infarction. ACE2 is also a negative regulator of Ang II-induced myocardial hypertrophy, fibrosis, and diastolic dysfunction. The ACE2-Ang 1-7/Mas axis may represent new possibilities for developing novel therapeutic strategies for the treatment of hypertension and cardiovascular diseases. In this review, we will summarize the biochemical and pathophysiological aspects of ACE2 with a focus on its role in diastolic and systolic heart failure. PMID- 21638103 TI - Efficacy of radiosynoviorthesis and its impact on chosen inflammatory markers. AB - Radiosynoviorthesis is used for the local treatment of recurrent joint effusions and leads to synovium necrosis after radionuclide administration. This procedure provides opportunity to full recovery of normal synovium function after local corticosteroids and systemic modifying drugs failure. PMID- 21638104 TI - Rational love, relational medicine: psychiatry and the accumulation of precarious kinship. AB - In north Indian psychiatry, clinical attentions to women's symptoms often involve scrutiny of emotions related to marriage and its breakdown. In pharmaceutically oriented practice, relations are used to evaluate biologies, and drugs produce the truth about relations at the same time that they produce the truth about bodies. In the process, clinical practice often involves unmaking relations, generating loss, in certain instances, as a dire result. In this, a particular kind of clinical knowing emerges, engaging broad cultural and historical connections between love and madness more than definitions of right and wrong unions. In asking how disciplinary and relational modes of biomedicine converge, I argue that in north Indian psychiatry's attentions to women, rather than enforcing normative configurations of "the family," biomedicine grapples with the gendered fallout of kinship. PMID- 21638105 TI - Role of serial order in the impact of talker variability on short-term memory: testing a perceptual organization-based account. AB - In two experiments, we examined the impact of the degree of match between sequential auditory perceptual organization processes and the demands of a short term memory task (memory for order vs. item information). When a spoken sequence of digits was presented so as to promote its perceptual partitioning into two distinct streams by conveying it in alternating female (F) and male (M) voices (FMFMFMFM)--thereby disturbing the perception of true temporal order--recall of item order was greatly impaired (as compared to recall of item identity). Moreover, an order error type consistent with the formation of voice-based streams was committed more quickly in the alternating-voice condition (Exp. 1). In contrast, when the perceptual organization of the sequence mapped well onto an optimal two-group serial rehearsal strategy--by presenting the two voices in discrete clusters (FFFFMMMM)--order, but not item, recall was enhanced (Exp. 2). The results are consistent with the view that the degree of compatibility between perceptual and deliberate sequencing processes is a key determinant of serial short-term memory performance. Alternative accounts of talker variability effects in short-term memory, based on the concept of a dedicated phonological short-term store and a capacity-limited focus of attention, are also reviewed. PMID- 21638106 TI - Are we aware of our ability to forget? Metacognitive predictions of directed forgetting. AB - It is often important to accurately predict not only what information we will later remember, but also what information we are likely to forget. The present research examined these abilities in the context of item-method directed forgetting, to determine whether people are aware of their strategic control of remembering and forgetting, as well as what cues are used when making metacognitive judgments. Participants studied words, each of which was followed by a cue to remember (R) or forget (F) the word for an upcoming test, and also made predictions of how likely they would be to later recall each word. When asked to recall all of the words, regardless of cue, both recall and predicted recall were fairly sensitive to the R or F instructions, despite some overconfidence. A similar and stronger pattern was found when words were assigned positive or negative point values as cues to remember or forget. These findings suggest that item-based cues to remember or forget information can be successfully utilized when making metacognitive judgments, and that people are fairly aware of the control they have over both remembering and forgetting information. PMID- 21638107 TI - List-wide control is not entirely elusive: evidence from picture-word Stroop. AB - Stroop interference is attenuated in mostly incongruent lists, as compared with mostly congruent ones. This finding is referred to as the list-wide proportion congruence effect. The traditional interpretation refers to the strategic biasing of attention via list-wide control. In mostly incongruent lists, attention is biased away from the irrelevant words, whereas in mostly congruent lists, words are more fully processed. According to the item-specific account, the list-wide proportion congruence effect reflects stimulus-driven mechanisms, and not list wide control. The unambiguous evidence available to date strongly favors the item specific account. Using a picture-word Stroop task, we demonstrate a list-wide proportion congruence effect for 50% congruent items that are embedded in mostly incongruent and mostly congruent lists. This novel finding illustrates that the list-wide proportion congruence effect is not entirely dependent on item-specific contributions and supports the list-wide control account. We discuss factors impacting the emergence of list-wide control in Stroop tasks. PMID- 21638108 TI - WISC-IV profile in high-functioning autism spectrum disorders: impaired processing speed is associated with increased autism communication symptoms and decreased adaptive communication abilities. AB - Changes in the Wechsler Intelligence Scales for Children-IV (WISC-IV) may affect the IQ profile characteristic of autism spectrum disorders (ASD). Moreover, the association of particular component cognitive abilities (unlike overall IQ) with symptomatology and adaptive functioning in ASD remains unclear. This archival study characterizes the WISC-IV IQ profile among 56 high-functioning (IQ > 70) children with ASD and correlates WISC-IV performance with ASD and ADHD symptomatology and adaptive functioning. The ASD WISC-IV profile included strengths on Matrix Reasoning and Similarities, weaknesses on Comprehension (which correlated negatively with social symptoms) and the subtests comprising the Processing Speed Index (Coding, Symbol Search). Processing speed task performance correlated negatively with communication symptoms and positively with communication abilities, indicating its importance to functional outcomes in ASD. PMID- 21638109 TI - Measuring change in social interaction skills of young children with autism. AB - Designing effective treatments for improving early social behaviors in autism has been identified as a critical research need. One barrier to drawing conclusions about optimal treatments for children with autism is the use of highly varied dependent measures in the treatment literature. Contributing to this is the absence of "gold standard" assessment batteries. This is particularly true for assessing changes in social interaction impairments in very young children with autism. This paper addresses this issue by reviewing variables important in the development and evaluation of assessment measures, discussing previous studies' choices of socially-related dependent measures, and the strengths, limitations, and research questions pertaining to them. It concludes with recommendations for measurement selection and future directions for research. PMID- 21638110 TI - Rhizosphere competent Pantoea agglomerans enhances maize (Zea mays) and chickpea (Cicer arietinum L.) growth, without altering the rhizosphere functional diversity. AB - Plant growth promoting Pantoea agglomerans NBRISRM (NBRISRM) was able to produce 60.4 MUg/ml indole acetic acid and solubilize 77.5 MUg/ml tri-calcium phosphate under in vitro conditions. Addition of 2% NaCl (w/v) in the media induced the IAA production and phosphate solubilization by 11% and 7%, respectively. For evaluating the plant growth promotory effect of NBRISRM inoculation a micro plot trial was conducted using maize and chickpea as host plants. The results revealed significant increase in all growth parameters tested in NBRISRM inoculated maize and chickpea plants, which were further confirmed by higher macronutrients (N, P and K) accumulation as compared to un-inoculated controls. Throughout the growing season of maize and chickpea, rhizosphere population of NBRISRM were in the range 10(7)-10(8) CFU/g soil and competing with 10(7)-10(9) CFU/g soil with heterogeneous bacterial population. Functional richness, diversity, and evenness were found significantly higher in maize rhizosphere as compared to chickpea, whereas NBRISRM inoculation were not able to change it, in both crops as compared to their un-inoculated control. To the best of our knowledge this is first report where we demonstrated the effect of P. agglomerans strain for improving maize and chickpea growth without altering the functional diversity. PMID- 21638111 TI - Application of flow cytometry for the identification of Staphylococcus epidermidis by peptide nucleic acid fluorescence in situ hybridization (PNA FISH) in blood samples. AB - Staphylococcus epidermidis is considered to be one of the most common causes of nosocomial bloodstream infections, particularly in immune-compromised individuals. Here, we report the development and application of a novel peptide nucleic acid probe for the specific detection of S. epidermidis by fluorescence in situ hybridization. The theoretical estimates of probe matching specificity and sensitivity were 89 and 87%, respectively. More importantly, the probe was shown not to hybridize with closely related species such as Staphylococcus aureus. The method was subsequently successfully adapted for the detection of S. epidermidis in mixed-species blood cultures both by microscopy and flow cytometry. PMID- 21638112 TI - A survey of culturable aerobic and anaerobic marine bacteria in de novo biofilm formation on natural substrates in St. Andrews Bay, Scotland. AB - This study reports a novel study of marine biofilm formation comprising aerobic and anaerobic bacteria. Samples of quartz and feldspar, minerals commonly found on the earth, were suspended 5 m deep in the North Sea off the east coast of St. Andrews, Scotland for 5 weeks. The assemblage of organisms attached to these stones was cultivated under aerobic and anaerobic conditions in the laboratory. Bacteria isolated on Marine Agar 2216 were all Gram-negative and identified to genus level by sequencing the gene encoding 16S rRNA. Colwellia, Maribacter, Pseudoaltermonas and Shewanella were observed in aerobically-grown cultures while Vibrio was found to be present in both aerobic and anaerobic cultures. The obligate anaerobic bacterium Psychrilyobacter atlanticus, a recently defined genus, was identified as a close relative of isolates grown anaerobically. The results provide valuable information as to the main players that attach and form de novo biofilms on common minerals in sea water. PMID- 21638113 TI - tdd8: a TerD domain-encoding gene involved in Streptomyces coelicolor differentiation. AB - The Streptomyces coelicolor genome contains 17 TerD domain-encoding genes (tdd genes) of unknown function. The proteins encoded by these genes have been presumed to be involved in tellurite resistance on the basis of their homology with the protein TerD of Serratia marcescens. To elucidate the role of a Tdd protein (Tdd8), both a deletion mutant for the corresponding gene tdd8 (SCO2368) and a recombinant strain over-expressing tdd8 were produced in S. coelicolor M145. The deletion mutant (Deltatdd8), like the wild strain, was not resistant to potassium tellurite. The deletion was not lethal but had a marked effect on differentiation. The deletion strain showed more rapid growth in liquid medium and produced long chains of short spores with a dense and non-spherical spore wall on agar plates. The strain over-expressing tdd8 had a growth delay in liquid medium and produced very few spores of irregular shapes and sizes on solid medium. The results of this study demonstrated that Tdd proteins might have a function other than tellurite resistance and this function seems to be of crucial importance for the proper development of the actinomycete S. coelicolor. PMID- 21638114 TI - Observations and insights about strengthening our soldiers SOS. AB - The Special Issue (June 2011) of the Journal of Clinical Psychology in Medical Settings titled Strengthening Our Soldiers (SOS) and Their Families: Contemporary Psychological Advances Applied to Wartime Problems revealed the following important concerns: 1) Who is at risk for psychological sequelae during and following service in the U.S. military? 2) How to deliver the best treatment for our soldiers and veterans with PTSD, Traumatic Brain Injury and Pain? 3) How to train the trainers? and 4) What are the current priorities for service delivery, research and funding? Assessment strategies and tools are provided to assist in identification of suicidal ideation and behaviors, alcohol abuse in spouses, posttraumatic stress disorders, depression, brain injuries and post-concussion syndrome, as well as positive growth experiences. Empirically validated Cognitive Processing and Prolonged Exposure treatments are described as are the empirical results already in evidence in our military populations. The innovative use of Virtual Reality and Telehealth applications is demonstrated in both army and naval settings for preparing and reducing trauma in affected soldiers. The Functional and Occupational Rehabilitation Treatment (FORT) Program and its role in returning function to injured soldiers with musculoskeletal pain and motion restrictions, while also leading to reductions of anxiety, depression and use of medical services, is described. A critique about providing service-research for children's reactions to their parent's deployments and family functioning during separation and reintegration is provided. The need for theoretical-empirical approaches to understanding pain-behavior, anxiety dysregulation as it impacts the brain function and structure is provided by experts in pain, neuropsychology, brain circuitry and anxiety management of multiple traumas. This final paper in SOS provides commentary on SOS and describes possible future implications of current psychological knowledge related to military personnel and their families. PMID- 21638115 TI - Health status, quality of life, residential stability, substance use, and health care utilization among adults applying to a supportive housing program. AB - Supportive housing, defined as subsidized housing in conjunction with site-based social services, may help improve the health and residential stability of highly disadvantaged individuals. This study examined changes in health status, quality of life, substance use, health care utilization, and residential stability among 112 homeless and vulnerably housed individuals who applied to a supportive housing program in Toronto, Canada, from December 2005 to June 2007. Follow-up interviews were conducted every 6 months for 18 months. Comparisons were made between individuals who were accepted into the program (intervention) and those who were wait-listed (usual care) using repeated-measures analyses. Individuals who were accepted into the housing program experienced significantly greater improvements in satisfaction with living situation compared with individuals in the usual care group (time, F(3,3,261) = 47.68, p < 0.01; group * time, F(3,3,261) = 14.60, p < 0.01). There were no significant differences in other quality of life measures, health status, health care utilization, or substance use between the two groups over time. Significant improvement in residential stability occurred over time, independent of assigned housing group (time, F(3,3,261) = 9.96, p < 0.01; group * time, F(3,3,261) = 1.74, p = 0.17). The ability to examine the effects of supportive housing on homeless individuals was limited by the small number of participants who were literally homeless at baseline and by the large number of participants who gained stable housing during the study period regardless of their assigned housing status. Nonetheless, this study shows that highly disadvantaged individuals with a high prevalence of poor physical and mental health and substance use can achieve stable housing. PMID- 21638116 TI - Sleep disturbance and risk behaviors among inner-city African-American adolescents. AB - Adolescents tend to experience more problems with sleep loss as a natural consequence of puberty, whereas teens from impoverished urban areas are likely to witness neighborhood violence and/or engage in risk behaviors that may affect sleep. Data from the Mobile Youth Survey, a longitudinal study of impoverished inner-city African-American adolescents (1998-2005; N = 20,716; age range = 9.75 19.25 years), were used to compare paired years of annual surveys elicited by questions about how sleep was affected when bad things happen to friends or family. Using a cross-lagged panel multivariate approach comparing reports for two sequential years and controlling for age/gender plus exposure to traumatic stress and violence, prior sleep disturbance was associated with carrying a knife/gun, brandishing a knife/gun, using a knife/gun, quick temperedness, warmth toward mother, worry, and belief in the neighborhood street code in the latter year. Conversely, seeing someone cut, stabbed, or shot, using alcohol, worry, and internalized anger were associated with sleep disturbance in a latter year. Although a limited measure of sleep disturbance was used, these findings support further research to examine sleep disturbance and risk behaviors among low-income adolescents. PMID- 21638117 TI - Quantifying urbanization as a risk factor for noncommunicable disease. AB - The aim of this study was to investigate the poorly understood relationship between the process of urbanization and noncommunicable diseases (NCDs) in Sri Lanka using a multicomponent, quantitative measure of urbanicity. NCD prevalence data were taken from the Sri Lankan Diabetes and Cardiovascular Study, comprising a representative sample of people from seven of the nine provinces in Sri Lanka (n = 4,485/5,000; response rate = 89.7%). We constructed a measure of the urban environment for seven areas using a 7-item scale based on data from study clusters to develop an "urbanicity" scale. The items were population size, population density, and access to markets, transportation, communications/media, economic factors, environment/sanitation, health, education, and housing quality. Linear and logistic regression models were constructed to examine the relationship between urbanicity and chronic disease risk factors. Among men, urbanicity was positively associated with physical inactivity (odds ratio [OR] = 3.22; 2.27-4.57), high body mass index (OR = 2.45; 95% CI, 1.88-3.20) and diabetes mellitus (OR = 2.44; 95% CI, 1.66-3.57). Among women, too, urbanicity was positively associated with physical inactivity (OR = 2.29; 95% CI, 1.64 3.21), high body mass index (OR = 2.92; 95% CI, 2.41-3.55), and diabetes mellitus (OR = 2.10; 95% CI, 1.58 - 2.80). There is a clear relationship between urbanicity and common modifiable risk factors for chronic disease in a representative sample of Sri Lankan adults. PMID- 21638118 TI - Frequency and timing of nonconvulsive status epilepticus in comatose post-cardiac arrest subjects treated with hypothermia. AB - BACKGROUND: Therapeutic hypothermia (TH) improves outcomes in comatose patients resuscitated from cardiac arrest. However, nonconvulsive status epilepticus (NCSE) may cause persistent coma. The frequency and timing of NCSE after cardiac arrest is unknown. METHODS: Review of consecutive subjects treated with TH and receiving continuous EEG (cEEG) monitoring between 8/1/2009 and 11/16/2010. Demographic data, survival, and functional outcome were prospectively recorded. Each cEEG file was analyzed using standard definitions to define NCSE. Data were analyzed using descriptive and nonparametric statistics. RESULTS: Mean age of the 101 subjects was 57 years (SD 15) with most subjects being male (N = 55, 54%) and experiencing out-of-hospital cardiac arrest (N = 78; 77%). Ventricular fibrillation was the initial cardiac rhythm in 39 (38%). All subjects received TH. Thirty subjects (30%) awoke at a median of 41 h (IQR 30, 61) after cardiac arrest. A total of 29/30 (97%) subjects surviving to hospital discharge were awake. Median interval from arrest to placement of cEEG was 9 h (IQR 6, 12), at which time the mean temperature was 33.9 degrees C. NCSE occurred in 12 (12%) subjects. In 3/12 (25%) subjects, NCSE was present when the cEEG recording began. In 4 subjects, NCSE occurred within 8 h of cEEG recording. One (8%) subject with NCSE survived in a vegetative state. CONCLUSIONS: NCSE is common in comatose post cardiac arrest subjects receiving TH. Most seizures occur within the first 8 h of cEEG recording and within the first 12 h after resuscitation from cardiac arrest. Outcomes are poor in those who experience NCSE. PMID- 21638119 TI - Comparison between cerebral tissue oxygen tension and energy metabolism in experimental subdural hematoma. AB - BACKGROUND: An experimental swine model (n = 7) simulating an acute subdural hematoma (ASDH) was employed (1) to explore the relation between the brain tissue oxygenation (PbtO(2)) and the regional cerebral energy metabolism as obtained by microdialysis, and (2) to define the lowest level of PbtO(2) compatible with intact energy metabolism. METHODS: ASDH was produced by infusion of 7 ml of autologous blood (infusion rate 0.5 ml/min) by a catheter placed subdurally. PbtO(2) and microdialysis probes were placed symmetrically in the injured ("bad side") and non-injured ("good-side") hemispheres. Intracranial pressure (ICP) was monitored in the "good-side." RESULTS: ICP, cerebral perfusion pressure (CPP), PbtO(2), glucose, lactate, pyruvate, lactate-pyruvate ratio (LP ratio), glutamate, and glycerol were recorded at baseline (60 min) and post trauma (360 min). After the creation of the ASDH, PbtO(2) decreased significantly in both the hemispheres (P < 0.001). No significant difference was found between the sides post trauma. The LP ratio, glutamate, and glycerol in the "bad-side" increased significantly over the "good-side" where the values remained within the normal limits. A PbtO(2) value below approximately 25 mmHg was found to be associated with disturbed energy metabolism in the "bad-side" but not in the "good-side." No correlation was found between the LP ratio and PbtO(2) in either hemisphere. CONCLUSIONS: PbtO(2) monitoring accurately describes tissue oxygenation but does not disclose whether the oxygen delivery is sufficient for maintaining cerebral energy metabolism. Accordingly, it may not be possible to define a threshold level for PbtO(2) below which energy failure and permanent tissue damage occurs. PMID- 21638120 TI - The comparison of acoustic and psychic parameters of subjective tinnitus. AB - We aim to assess the correlation between audiometric data, and psychotic and acoustic measures associated with subjective tinnitus (ST) and to clarify the importance of the psychological process in determining the degree of subjective annoyance and disability due to tinnitus. Fifty-four patients experiencing unilateral ST were allocated for the study. Acoustic assessment of patients including LDL (loudness discomfort levels), MML (minimum masking level) and RI (residual inhibition) was performed. Tinnitus Handicap Inventory (THI), Beck Depression Inventory (BDI) and Visual Analog Scale (VAS) tests were performed for the psychological aspects of subjective annoyance. RI was positive in 23 patients with 13 frequency-matched stimuli at 8,000 Hz. Masking treatment response was successful in 16 RI-positive patients. Mean and standard deviation (SD) of THI scores were 38.77 +/- 23.63. Ten patients (%18.51) with tinnitus had >= 17 points score, which was significant for BDI. Mean and SD were 5.01 +/- 2.31 for VAS-1 scores (severity of tinnitus), 7.98 +/- 2.79 for VAS-2 (frequency and duration of tinnitus), 5.77 +/- 2.72 for VAS-3 (discomfort level), 3.56 +/- 3.30 for VAS-4 (attention deficit) and 3.31 +/- 3.31 for VAS-5 (sleep disorders). A significant correlation was found between the tinnitus duration time, age, gender and THI scores (P < 0.05). There were statistically significant correlations between VAS 1, 2, 3 scores and LDL, MML and RI (P > 0.05). RI might be largely frequency dependent and was found as an indicator for the masking treatment response. We did not notice statistically significant correlations between audiometric data and THI and BDI. There were correlations between with VAS and LDL and with MML and RI. VAS was simpler and easier for the assessment of ST. We should consider the psychological aspects of ST and assess it as a symptom separately with acoustic and psychotic tests. PMID- 21638121 TI - Vascular normalization: a real benefit? AB - It is well established that antibodies to vascular endothelial growth factor (VEGF) in combination with chemotherapeutic agents produce synergistic cytotoxicity in a range of cancer. In this review article, it has been analyzed whether the so-called vascular normalization of abnormal tumor blood vessels as an effect of VEGF inhibition in association with chemotherapeutic agents in the treatment of tumors produces a real benefit. Literature data show that the process of normalization of the structure of tumor blood vessels is not always accompanied with a real benefit. In fact as in the case of cerebral tumors, the process of normalization may induce a re-establishment of the low permeability characteristics of normal brain microvasculature, preventing the delivery of chemotherapeutics. PMID- 21638122 TI - Preclinical anti-angiogenesis and anti-tumor activity of SIM010603, an oral, multi-targets receptor tyrosine kinases inhibitor. AB - OBJECTIVE: SIM010603 is a structurally novel, oral, multi-targeted receptor tyrosine kinase inhibitor. This study investigated the anti-angiogenic and anti tumor effects of SIM010603. METHODS: A radiometric protein kinase assay was used for measuring the kinase activity of the 32 protein kinases. Receptor phosphorylation was determined by enzyme-linked immunosorbent assay (ELISA). Cell proliferation was measured by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Cell chemotaxis was evaluated by modified Boyden chamber assay. Effect of SIM010603 on angiogenesis was examined by mouse cornea angiogenesis assay. Effect of SIM010603 on xenografts was assessed by tumor growth delay. Effects of SIM010603 on tumor microvascular density (MVD), recruitment of pericytes, and pericyte encapsulation of tumor vessels were analyzed by immunofluorescent staining technique. RESULTS: SIM010603 inhibited stem cell factor receptor (Kit), vascular endothelial growth factor receptor-2 (VEGFR-2), platelet-derived growth factor receptor-beta (PDGFR-beta), glial cell line-derived neurotrophic factor receptor (Rearranged during Transfection; RET), and Fms-like tyrosine kinase-3 (FLT3) with IC(50) values between 5.0 and 68.1 nmol/l. SIM010603 inhibited the phosphorylation of PDGFR-beta and VEGFR-2. Moreover, SIM010603 inhibited endothelial cell proliferation, endothelial cells chemotaxis, and corneal angiogenesis. Although SIM010603 exhibited lower activity in regard to proliferation of NCI-H460, MDA-MB-435, and T241-VEGF-A cells (IC(50) > 1 MUmol/l), SIM010603 inhibited tumor growth in these xenograft tumor growth models. SIM010603 reduced tumor MVD in T241-VEGF-A tumor xenograft models and decreased positive signals of CD31, NG2 in MDA-MB-435, and LLC-SW-44 xenograft tumor models. CONCLUSIONS: These results support the clinical assessment of SIM010603 as a therapeutic agent for cancer. PMID- 21638123 TI - A Phase I study to assess the safety, tolerability, and pharmacokinetics of AZD4877, an intravenous Eg5 inhibitor in patients with advanced solid tumors. AB - PURPOSE: Inhibition of kinesin spindle protein or Eg5 causes the formation of monoastral mitotic spindles, which leads to cell death. AZD4877 is a specific, potent inhibitor of Eg5. METHODS: This was a Phase I, open-label, two-part study to evaluate the maximum tolerated dose (MTD) and safety and tolerability of AZD4877 in patients with advanced solid malignancies. In part A, the MTD of AZD4877, administered as three weekly 1-h intravenous (iv) infusions in a 28-day schedule, was determined by evaluating dose-limiting toxicity (DLT). In part B, the safety, tolerability, and pharmacokinetic profile of AZD4877 at the MTD were evaluated. RESULTS: In part A, 29 patients received at least one dose of AZD4877 (5 mg, n = 4; 7.5 mg, n = 4; 10 mg, n = 3; 15 mg, n = 3; 20 mg, n = 3; 30 mg, n = 6; 36 mg, n = 3; 45 mg, n = 3). The MTD was defined as 30 mg, with the primary DLT being neutropenia. Although exposures appeared to be similar at the AZD4877 20 and 30 mg doses, dose reductions and omissions were higher in the 30-mg cohort; therefore, an intermediate dose, 25 mg, was evaluated in part B (n = 14). In part B, neutropenia remained the most commonly reported causally related adverse event. Exposure to AZD4877 was approximately dose proportional. Severity of neutropenia was related to exposure. CONCLUSION: The MTD of AZD4877 given as a 1-h iv infusion on days 1, 8, and 15 of a 28-day cycle was 30 mg. At the selected 25 mg dose, AZD4877 had an acceptable safety profile. PMID- 21638124 TI - Immuno-cell therapy with antecedent surgery has superior actuarial survival to immuno-cell therapy without antecedent surgery for advanced cancers. AB - BACKGROUND: Immuno-cell therapy using activated lymphocytes (ALs) and/or dendritic cells (DCs) is considered one of the less toxic supportive therapies compared with conventional chemotherapy and radiotherapy, especially for the treatment for advanced cancers. To improve the efficacy of immuno-cell therapy for such cancer, clinical data were analyzed in this preliminary study. PATIENTS AND METHODS: The clinical data of 38 consecutive patients with advanced cancer who underwent at least one course of treatment with ALs and/or matured DCs, with or without antecedent surgery or additional conventional chemotherapy and/or radiotherapy, were evaluated. RESULTS: Of the 23 patients who received surgery before immuno-cell therapy, 2 (8.7%) showed a complete response (CR) and 15 (65%) showed a partial response (PR) or prolonged stable disease (SD). Of the 15 remaining patients who did not undergo antecedent surgery, there was no CR but 7 (46%) showed PR or prolonged SD. Actuarial survival is one of the important indices for the evaluation of anticancer therapies that present longer durable efficacy of immunotherapy compared with conventional anticancer chemotherapy and radiotherapy, and actuarial survival analysis revealed that immuno-cell therapy with antecedent surgery afforded significantly longer survival than immuno-cell therapy without antecedent surgery (P < 0.001). CONCLUSION: Antecedent surgical resection of tumors is advisable for obtaining better efficacy of immuno-cell therapy, even in advanced cancer patients. PMID- 21638125 TI - Ectonucleotidases CD39 and CD73 on OvCA cells are potent adenosine-generating enzymes responsible for adenosine receptor 2A-dependent suppression of T cell function and NK cell cytotoxicity. AB - The ectonucleotidases CD39 and CD73 degrade immune stimulatory ATP to adenosine that inhibits T and NK cell responses via the A(2A) adenosine receptor (ADORA2A). This mechanism is used by regulatory T cells (T(reg)) that are associated with increased mortality in OvCA. Immunohistochemical staining of human OvCA tissue specimens revealed further aberrant expression of CD39 in 29/36 OvCA samples, whereas only 1/9 benign ovaries showed weak stromal CD39 expression. CD73 could be detected on 31/34 OvCA samples. While 8/9 benign ovaries also showed CD73 immunoreactivity, expression levels were lower than in tumour specimens. Infiltration by CD4(+) and CD8(+) T cells was enhanced in tumour specimens and significantly correlated with CD39 and CD73 levels on stromal, but not on tumour cells. In vitro, human OvCA cell lines SK-OV-3 and OaW42 as well as 11/15 ascites derived primary OvCA cell cultures expressed both functional CD39 and CD73 leading to more efficient depletion of extracellular ATP and enhanced generation of adenosine as compared to activated T(reg). Functional assays using siRNAs against CD39 and CD73 or pharmacological inhibitors of CD39, CD73 and ADORA2A revealed that tumour-derived adenosine inhibits the proliferation of allogeneic human CD4(+) T cells in co-culture with OvCA cells as well as cytotoxic T cell priming and NK cell cytotoxicity against SK-OV3 or OAW42 cells. Thus, both the ectonucleotidases CD39 and CD73 and ADORA2A appear as possible targets for novel treatments in OvCA, which may not only affect the function of T(reg) but also relieve intrinsic immunosuppressive properties of tumour and stromal cells. PMID- 21638126 TI - Low molecular weight hyaluronan preconditioning of tumor-pulsed dendritic cells increases their migratory ability and induces immunity against murine colorectal carcinoma. AB - We have recently shown that systemic administration of low molecular weight hyaluronan (LMW HA) significantly reduces colorectal carcinoma (CRC) growth in vivo. The elicited response is partially mediated by activated dendritic cells (DC). To potentiate the ability of DC loaded with whole tumor lysate (DC/TL) to induce immunity against CRC in mice, we aimed to study the effects of preconditioning DC with LMW HA for therapeutic vaccination. LMW HA improved maturation of ex vivo generated DC, increased IL-12, decreased IL-10 production, and enhanced a MLR activity in vitro. Although TNF-alpha showed a similar capacity to mature DC, preconditioning of DC/TL with LMW HA increased their ability to migrate in vitro toward CCL19 and CCL-21 in a CD44- and a TLR4 independent manner; this effect was superior to Poly(I:C), LPS, or TNF-alpha and partially associated with an increase in the expression of CCR7. Importantly, LMW HA dramatically enhanced the in vivo DC recruitment to tumor-regional lymph nodes. When these LMW HA-treated CRC tumor lysate-pulsed DC (DC/TL/LMW HA) were administered to tumor-bearing mice, a potent antitumor response was observed when compared to DC pulsed with tumor lysate alone and matured with TNF-alpha. Then, we showed that splenocytes isolated from animals treated with DC/TL/LMW HA presented a higher proliferative capacity, increased IFN-gamma production, and secreted lower levels of the immunosuppressive IL-10. Besides, increased specific CTL response was observed in DC/TL/LMW HA-treated animals and induced long-term protection against tumor recurrence. Our data show that LMW HA is superior to other agents at inducing DC migration; therefore, LMW HA could be considered a new adjuvant candidate in the preparation of DC-based anticancer vaccines with potent immunostimulatory properties. PMID- 21638127 TI - Skin tumor responsiveness to interleukin-2 treatment and CD8 Foxp3+ T cell expansion in an immunocompetent mouse model. AB - Recombinant human interleukin-2 (rhIL-2) therapy is approved for treating patients with advanced melanoma yet significant responses are observed in only 10 15% of patients. Interleukin-2 induces Foxp3 expression in activated human CD8 T cells in vitro and expands circulating CD8 Foxp3+ T cells in melanoma patients. Employing IL-2 responsive (B16-F1, B16-BL6, JB/MS, MCA-205) and nonresponsive (JB/RH, B16-F10) subcutaneous tumor mouse models, we evaluated CD8 Foxp3+ T cell distribution and changes in response to rhIL-2 (50,000 U, i.p. or s.q., twice daily for 5 days). In tumor-free mice and subcutaneous tumor-bearing mouse models, CD8 Foxp3+ T cells were a rare but naturally occurring cell subset. Primarily located in skin-draining lymph nodes, CD8 Foxp3+ T cells expressed both activated T cell (CD28(+), CD44(+)) and Treg (CTLA4(+), PD1(lo/var), NKG2A(+/var)) markers. Following treatment with rhIL-2, a dramatic increase in CD8 Foxp3+ T cell prevalence was observed in the circulation and tumor-draining lymph nodes (TD.LNs) of animals bearing IL-2 nonresponsive tumors, while no significant changes were observed in the circulation and TD.LNs of animals bearing IL-2 responsive tumors. These findings suggest expansion of CD8 Foxp3+ T cell population in response to rhIL-2 treatment may serve as an early marker for tumor responsiveness to immunotherapy in an immune competent model. Additionally, these data may provide insight to predict response in patients with melanoma undergoing rhIL-2 treatment. PMID- 21638128 TI - CCL5, CXCL16, and CX3CL1 are associated with Henoch-Schonlein purpura. AB - Chemokines are involved in the pathogenesis of various vascular inflammations. However, information about chemokines in Henoch-Schonlein purpura (HSP) is limited. Herein, we investigated the serum CCL5, CXCL16, and CX3CL1 levels in HSP patients with controls and the ability of sera from HSP patients on chemokine production in human dermal microvascular endothelial cells. Enzyme-linked immunosorbent assay (ELISA) detected serum CCL5, CXCL16, and CX3CL1 levels in patients with HSP. Human dermal microvascular endothelial cell line (HMEC-1) was treated with sera from patients with HSP at different stages, patients with acute spontaneous urticaria, or controls. Serum levels of CCL5, CXCL16, and CX3CL1 were elevated in HSP patients at acute stage, which correlated with the severity of this disease. Sera from patients with active HSP markedly induced CCL5, CXCL16, and CX3CL1 production at both mRNA and protein levels. In addition, patients' sera-stimulated HMEC-1 supernatants enhanced HL-60 or THP-1 cells migration. Furthermore, patients' sera increased the phosphorylation of inhibitor of kappaB alpha (IkappaBalpha) and phosphorylation of extracellular signal-regulated kinase (ERK)1/2 protein levels, upregulated the translocation of nuclear factor-kappaB (NF-kappaB) p65 to the nucleus. Taken together, we show firstly that CCL5, CXCL16, and CX3CL1 may be involved in the pathogenesis of HSP. Factors present in sera from patients with active HSP may act as an inducer of inflammatory response in HMEC-1 cells and contribute to chemokine production through NF-kappaB and ERK 1/2 pathways. PMID- 21638129 TI - Quantitative EEG in type 1 diabetic adults with childhood exposure to severe hypoglycaemia: a 16 year follow-up study. AB - AIMS/HYPOTHESIS: In diabetic children and adolescents, a history of severe hypoglycaemia (SH) has been associated with increased slow EEG activity and reduced cognition, possibly due to harmful effects of SH on the developing brain. In a group of type 1 diabetic patients with early exposure to SH, who had EEG abnormalities and reduced cognition in childhood, we have recently demonstrated that the reduced cognition may persist into adulthood. We have now assessed whether the reduced cognition was accompanied by lasting EEG abnormalities. METHODS: In 1992-1993, we studied EEG and cognition in 28 diabetic children and 28 matched controls. 16 years later, we re-investigated the same participants, with 96% participation rate. Diabetic participants were classified as with (n = 9) or without (n = 18) early SH, defined as episodes with convulsions or loss of consciousness by 10 years of age. For each EEG band (delta, theta, alpha and beta) and cerebral region (frontocentral, temporal, and parietooccipital), we calculated relative amplitudes and amplitude asymmetry. We also calculated occipital alpha mean frequency, alpha peak frequency at maximum amplitude, alpha peak width, and theta regional mean frequencies. We examined whether these EEG measures, relative to age- and sex-matched controls, differed between diabetic participants with and without early SH. RESULTS: We found no association of early SH with any of the EEG measures. CONCLUSIONS/INTERPRETATION: Childhood SH was not associated with EEG abnormalities in young type 1 diabetic adults. Our findings suggest that the reduced adulthood cognition associated with childhood exposure to SH is not accompanied by lasting EEG abnormalities. PMID- 21638130 TI - Individual and cumulative effect of type 2 diabetes genetic susceptibility variants on risk of coronary heart disease. AB - AIMS/HYPOTHESIS: Type 2 diabetes is a major risk factor for CHD. We hypothesised that diabetes genetic susceptibility variants might be associated with increased CHD risk. METHODS: We examined the individual and cumulative effect of 38 common genetic variants previously reported to be associated with type 2 diabetes on risk of incident CHD in 20,467 participants of the European Prospective Investigation into Cancer and Nutrition (EPIC) Norfolk Study who had been free of CHD at baseline. RESULTS: During a mean follow-up of 10.7 years, 2,190 participants had a CHD event. Two individual variants next to the TSPAN8 (HR 1.07, 95% CI 1.00-1.14) and the CDKN2A/B region (1.11, 1.04-1.17) were significantly associated with increased CHD risk. A genetic score based on the 38 diabetes variants was significantly associated with an increased risk of CHD (1.08, 1.01-1.14 per score tertile). Adjustment for prevalent and incident diabetes attenuated the association of the TSPAN8 variant (1.06, 0.99-1.13) and the genetic score (1.05, 0.99-1.12 per score tertile) with CHD risk, but not that of the CDKN2A/B variant (1.11, 1.05-1.18). Addition of the genetic score did not improve risk discrimination based on clinical risk factors. CONCLUSIONS/INTERPRETATION: The increased risk of CHD observed with genetic susceptibility to type 2 diabetes was at least partly mediated by its diabetes predisposing effect and was not useful for clinical risk discrimination. The potential role of pathways associated with the variant CDKN2A/B in linking diabetes and CHD needs further exploration. PMID- 21638131 TI - Associations between single-nucleotide polymorphisms (+45T>G, +276G>T, -11377C>G, -11391G>A) of adiponectin gene and type 2 diabetes mellitus: a systematic review and meta-analysis. AB - AIMS/HYPOTHESIS: The associations between adiponectin polymorphisms and type 2 diabetes have been studied widely; however, results are inconsistent. METHODS: We searched electronic literature databases and reference lists of relevant articles. A fixed or random effects model was used on the basis of heterogeneity. Sub-group and meta-regression analyses were conducted to explore the sources of heterogeneity. RESULTS: There were no statistically significant associations between +45T>G (rs2241766), +276G>T (rs1501299), -11391G>A (rs17300539) and type 2 diabetes risk. However, for -11377C>G (rs266729), the pooled OR (95% CI) for G vs C allele was 1.07 (1.03-1.11, p = 0.001). Subgroup analysis by study design revealed that -11377C>G (rs266729) dominant model (CG+GG vs CC, p = 0.0008) and G vs C allele (p = 0.0004) might be associated with type 2 diabetes risk in population-based case-control studies. After stratification by ethnicity, we found that -11377C>G (rs266729) dominant model (CG+GG vs CC, p = 0.004) and G vs C allele (p = 0.001) might be associated with type 2 diabetes risk in white individuals. In individuals with a family history of diabetes, the presence of 11391G>A (rs17300539) dominant model (GA+AA vs GG) and A vs G allele might be associated with increased risk of type 2 diabetes. CONCLUSIONS/INTERPRETATION: The presence of +45T>G (rs2241766), +276G>T (rs1501299) and -11391G>A (rs17300539) do not appear to influence the development of type 2 diabetes. However, G vs C allele of -11377C>G (rs266729) might be a risk factor for type 2 diabetes. PMID- 21638132 TI - Diabetic retinopathy in type 1 diabetes-a contemporary analysis of 8,784 patients. AB - AIMS/HYPOTHESIS: The aim of this study was to analyse the risk profile for diabetic retinopathy under real-life conditions in a large cohort of patients with type 1 diabetes. METHODS: Patients (n = 18,891) with childhood, adolescent or adult onset of type 1 diabetes from the prospective German Diabetes Documentation System survey were analysed. A total of 8,784 patients fulfilled the inclusion criterion, which was availability of retinopathy status. Retinopathy grading (any retinopathy, advanced retinopathy), treatment regimens and risk factors were prospectively recorded and tested as covariates by Kaplan Meier analysis and logistic regression. RESULTS: Any retinopathy was present in 27.4% and advanced retinopathy (severe non-proliferative or proliferative diabetic retinopathy) in 8.0% of the cohort. After 40 years of diabetes, the cumulative proportion of patients with any retinopathy and advanced retinopathy was 84.1% and 50.2%, respectively. In multiple regression analysis, risk factors for any retinopathy were diabetes duration (OR 1.167 per year), HbA(1c) >7.0% (53 mmol/mol) (OR 2.225), smoking (OR 1.295) and male sex (OR 1.187) (p < 0.0001 for all). Young age at onset (5 vs 15 years at disease onset) was protective (0.410, p < 0.0001). No glycaemic threshold was detected for retinopathy protection. Risk factors for advanced retinopathy were duration (1.124 per year, p < 0.0001), male sex (1.323, p = 0.0020), HbA(1c) >7.0% (53 mmol/mol) (1.499, p < 0.0001), triacylglycerol >1.7 mmol/l (1.398, p = 0.0013) and blood pressure >140/90 mmHg (1.911, p < 0.0001). CONCLUSIONS/INTERPRETATION: The prevalence of retinopathy remains significant in type 1 diabetes. Any improvement of metabolic control and non-smoking is protective, while hypertension affects progression to severe levels under real-life conditions. These data reinforce the validity of multifactorial concepts for morbidity protection in type 1 diabetes. PMID- 21638133 TI - Screening for diabetes using an oral glucose tolerance test within a western multi-ethnic population identifies modifiable cardiovascular risk: the ADDITION Leicester study. AB - AIMS/HYPOTHESIS: The aim of this study was to determine the frequency of undiagnosed glucose abnormalities and the burden of cardiovascular disease (CVD) risk among south Asians and white Europeans attending a systematic screening programme for type 2 diabetes (ADDITION-Leicester) and to estimate the achievable risk reduction in individuals identified with glucose disorders. METHODS: Random samples of individuals (n = 66,320) from 20 general practices were invited for a 75 g OGTT and CVD risk assessment. Ten-year CVD risk among screen-detected people with diabetes or impaired glucose regulation (IGR) (impaired fasting glycaemia and/or impaired glucose tolerance [IGT]) was computed using the Framingham-based ETHRISK engine and achievable risk reduction was predicted using relative reductions for treatments extracted from published trials. RESULTS: A total of 6,041 participants (48% male, 22% south Asian) aged 40-75 years inclusive were included. Undiagnosed glucose disorders occurred more frequently in south Asians than white Europeans; age and sex adjusted odds ratios were 1.74 (95% CI 1.42 2.13) and 2.30 (95% CI 1.68-3.16) for IGT and diabetes respectively. Prevalence of any undetected glucose disorder was 17.5% in the whole cohort. Adjusted 10 year risk was similar in screen-detected people with IGR and diabetes (18.3% vs 21.6%), and was higher in south Asians across the glucose spectrum. Absolute CVD risk reductions of up to 13% in those with screen-detected type 2 diabetes and 6% in IGR are achievable using existing cardioprotective therapies. CONCLUSIONS/INTERPRETATION: Population screening with an OGTT identifies a significant burden of modifiable CVD risk, especially within south Asian groups. Strategies enticing this population to consider screening programmes are urgently needed as significant risk reduction is possible once a glucose abnormality is identified. TRIAL REGISTRATION: ClinicalTrials.gov NCT00318032. FUNDING: The project is funded for support and treatment costs by NHS Department of Health Support for Science and project grants. PMID- 21638134 TI - Spontaneous gelation of a novel histamine H4 receptor antagonist in aqueous solution. AB - PURPOSE: Low molecular weight hydrogelators typically require a stimulus such as heat, antisolvent, or pH adjustment to produce a gel. This study examines gelation of a novel histamine H4 receptor antagonist that forms hydrogels spontaneously at room temperature. METHODS: To elucidate the mechanism and structural moieties responsible for this unusual gelation, hydrogels were characterized by rheology, optical microscopy, and XRD. SEM was performed on xerogels; NMR measurements were conducted in gelator solutions in the presence of a gel-breaker. The influence of temperature, concentration, pH, and ionic strength on elastic and viscous moduli of the hydrogels was evaluated; gel points were established via thorough rheological criteria. RESULTS: The observed are "true" gels with a fibrillar texture and lamellar microstructure. On a molecular level, the gels are composed of aggregates of partially ionized species stabilized by hydrophobic interactions of aromatic moieties. The gel-to-sol transition occurs at physiologically relevant temperatures and is concentration-, pH-, and ionic strength-dependent. CONCLUSIONS: We hypothesize that this spontaneous gelation is due to the so-called "spring" effect, a high energy salt form that transiently increases aqueous solubility above its equilibrium limit. Upon equilibration, this supersaturated system undergoes aggregation that avoids crystallization and produces a hydrogel. PMID- 21638135 TI - siRNA-mediated down-regulation of P-glycoprotein in a Xenograft tumor model in NOD-SCID mice. AB - PURPOSE: The efficacy of chemotherapy is decreased due to over-expression of the drug transporter P-glycoprotein (P-gp). This study was conducted to determine the feasibility of down-regulating tumor P-gp levels with non-viral siRNA delivery in order to sensitize the tumors to drug therapy. METHODS: P-gp over-expressing MDA435/LCC6 MDR1 cells were used to establish xenografts in NOD-SCID mouse. Cationic polymers polyethylenimine (PEI) and stearic acid-substituted poly-L lysine (PLL-StA) were formulated with P-gp- specific siRNAs and delivered intratumorally to explore the feasibility of P-gp down-regulation in tumors. Intravenous DoxilTM was administered to investigate tumor growth. RESULTS: PEI and PLL-StA effectively delivered siRNA to MDA435/LCC6 MDR1 cells in vitro to reduce P-gp expression for 3 days. Intratumoral injection of siRNA with the carriers resulted in 60-80% and 20-32% of siRNA retention in tumors after 24 and 96 hr, respectively. This led to ~29.0% and ~61.5% P-gp down-regulation with PEI- and PLL-StA-mediated siRNA delivery, respectively. The P-gp down-regulation by intratumoral siRNA injection led to better response to systemic DoxilTM treatment, resulting in slowed tumor growth in originally doxorubicin-resistant tumors. CONCLUSION: Effective P-gp down-regulation was feasible with polymeric siRNA delivery in a xenograft model, resulting in an enhanced response to the drug therapy. PMID- 21638136 TI - Novel therapies targeting inner mitochondrial membrane--from discovery to clinical development. AB - Mitochondrial oxidative stress and dysfunction have been implicated in the aging process and in numerous chronic diseases. The need for therapies that can protect and/or improve mitochondrial function is obvious. However, the development of mitoprotective drugs has been hampered by a number of challenges, and there are at present no approved therapies for mitochondrial dysfunction. This article describes the original discovery, preclinical development, and clinical development of a novel class of small peptide molecules that selectively target the inner mitochondrial membrane and protect mitochondrial function. These compounds have the potential to be a paradigm-shifting approach to the treatment of mitochondrial dysfunction, which underlies many common diseases, including cardiorenal, neurologic, and metabolic disorders. PMID- 21638137 TI - Crystallization of organic glasses: effects of polymer additives on bulk and surface crystal growth in amorphous nifedipine. AB - PURPOSE: To study the influence of polymer additives on bulk and surface crystal growth in organic glasses (amorphous solids), which are being investigated for delivering poorly soluble drugs and in this role must resist crystallization. Recent studies have discovered new modes of crystal growth that emerge as organic liquids are cooled to form glasses: one existing in the bulk (GC growth) and another at the surface, both leading to crystal growth much faster than predicted by standard theories. METHODS: Bulk and surface crystal growth rates were measured in nifedipine glasses doped with polyvinylpyrrolidone (PVP) of different molecular weights. AFM enabled observation of the microstructure of surface growing crystals. RESULTS: Polymer additives influence bulk and surface crystal growth differently. For every weight percent of PVP added, surface crystal growth of nifedipine slows by two times at 12 degrees C below T (g), whereas bulk crystal growth slows by 10 times. In contrast to the polymers, the VP dimer had little effect on crystal growth. CONCLUSIONS: Polymer additives inhibit crystal growth in nifedipine glasses more strongly in the bulk than at the surface. The effectiveness of crystallization inhibitors depends not only on intermolecular interactions but also on molecular sizes. PMID- 21638138 TI - Intravenous thrombolytics for ischemic stroke. AB - For many decades, intravenous (IV) thrombolytics have been delivered to treat acute thrombosis. Although these medications were originally effective for coronary thrombosis, their mechanisms have proven beneficial for many other disease processes, including ischemic stroke. Treatment paradigms for acute ischemic stroke have largely followed those of cardiology. Specifically, the aim has been to recanalize the occluded artery and to restore perfusion to the brain that remains salvageable. To that end, rapid clot lysis was sought using thrombolytic medicines already proven effective in the coronary arteries. IV thrombolysis for ischemic stroke began its widespread adoption in the late 1990s after the publication of the National Institute of Neurological Disorders and Stroke study. Since that time, other promising IV-thrombolytics have been developed and tested in human trials, but as of yet, none have been proven better than a placebo. Adjunctive treatments are also being evaluated. The challenge remains balancing reperfusion and salvaging brain tissue with the potential risks of brain hemorrhage. PMID- 21638140 TI - High-resolution measurement of the unsteady velocity field to evaluate blood damage induced by a mechanical heart valve. AB - We investigate the potential of prosthetic heart valves to generate abnormal flow and stress patterns, which can contribute to platelet activation and lysis according to blood damage accumulation mechanisms. High-resolution velocity measurements of the unsteady flow field, obtained with a standard particle image velocimetry system and a scaled-up model valve, are used to estimate the shear stresses arising downstream of the valve, accounting for flow features at scales less than one order of magnitude larger than blood cells. Velocity data at effective spatial and temporal resolution of 60 MUm and 1.75 kHz, respectively, enabled accurate extraction of Lagrangian trajectories and loading histories experienced by blood cells. Non-physiological stresses up to 10 Pa were detected, while the development of vortex flow in the wake of the valve was observed to significantly increase the exposure time, favouring platelet activation. The loading histories, combined with empirical models for blood damage, reveal that platelet activation and lysis are promoted at different stages of the heart cycle. Shear stress and blood damage estimates are shown to be sensitive to measurement resolution. PMID- 21638141 TI - Hyperinsulinemia may promote growth without GH in children after resection of suprasellar brain tumors. AB - It remains elusive what factors induce growth without growth hormone (GWGH) in children after neurosurgery of brain tumors. Growth velocity and endocrinological data were compared between the patients with and without GWGH. We experienced three patients with GWGH (median, 12 years; 2 germinoma and 1 craniopharyngioma; three females; group 1) and 11 patients without (12 years; 8 craniopharyngioma, 2 germinoma and 1 medulloblastoma; 7 males; group 2) after neurosurgery. All patients in group 2 received GH replacement therapy. Growth velocity and endocrinological data were compared. Median height velocity was normal in group 1 (5.5 cm/year), but low in group 2 (2.2 cm/year), which improved after GH replacement therapy (7.0 cm/year). Median serum insulin level was increased in group 1 (87.0 MUU/ml, P < 0.05) compared with normal level in group 2 (10.0 MUU/ml). Despite hyperinsulinemia, serum glucose level was normal in group 1. Three of 5 with hyperinsulinemia and 2 of 9 without were obese or overweight, but the difference was not significant. Current body mass index and serum levels of IGF-1, IGFBP-3, leptin, and prolactin were similar between groups. Serum estradiol was prepuberty level in group 1. Hyperinsulinemia may induce GWGH in children with brain tumors after neurosurgery. PMID- 21638139 TI - Prevention of cardioembolic stroke. AB - Cardiac causes of ischemic stroke lead to severe neurological deficits from large intracranial artery occlusion compared to small vessel ischemic stroke. The most common cause of cardioembolic stroke is atrial fibrillation (AF), which has an increasing incidence with age. AF stroke trials demonstrate that anti-coagulation is superior to anti-platelet therapy in terms of ischemic stroke prevention. Recently, warfarin was compared with dabigatran, an oral, direct thrombin inhibitor, and was found to be at least equally effective in reducing ischemic stroke with less intracranial bleeding risk. Future research is investigating other direct thrombin inhibitors as potential alternatives to warfarin, which has a narrow therapeutic index, requires frequent blood monitoring, has multiple drug interactions, and a higher rate of intracranial bleeding. Other causes of cardioembolic stroke include myocardial infarction, left ventricular thrombus, reduced ejection fraction, valvular abnormalities, and endocarditis. Patent foramen ovale is a common finding on echocardiograms in patients with and without stroke (up to 20% of the population), and it is a controversial source of cryptogenic stroke. The best way to prevent cardioembolic stroke remains early detection and treatment of AF, and treating the underlying stroke mechanism. Cardiac magnetic resonance imaging is an emerging technology and reveals some sources of cardiac embolism missed by echocardiography, and might provide an additional diagnostic tool in investigating cardioembolic stroke. PMID- 21638142 TI - Coexistence of spinal schwannoma with unusual malignant peripheral T-cell lymphoma within a lumbar spine lesion. PMID- 21638143 TI - A simple and safe system for avoiding electrode dislodgement in patients undergoing peripheral nerve field stimulation for chronic pain. PMID- 21638144 TI - Improvement of hand dexterity induced by stimulation of the peduncolopontine nucleus in a patient with advanced Parkinson's disease and previous long-lasting bilateral subthalamic DBS. AB - We report the case of a patient already submitted to bilateral deep-brain stimulation (DBS) of the subthalamic nucleus (STN) who started to develop gait impairment, postural imbalance and frequent falls in the course of the disease and who subsequently underwent DBS of the right pedunculopontine nucleus (PPN) at our institute. An immediate clinical benefit in hand dexterity was observed with acute external stimulation and maintained after the definitive implant of the internal pulse generator (IPG) at 6 months' follow-up. The benefit on hand dexterity seemed to be related to the interactions between the PPN low-frequency stimulation and the bilateral STN high-frequency stimulation. PMID- 21638145 TI - Percutaneous transhepatic removal of bile duct stones: results of 261 patients. AB - PURPOSE: To determine the effectiveness of percutaneous transhepatic removal of bile duct stones when the procedure of endoscopic therapy fails for reasons of anatomical anomalies or is rejected by the patient. METHODS: Between April 2001 and May 2010, 261 patients (138 male patients and 123 female patients; age range, 14-92 years; mean age, 64.6 years) with bile duct stones (common bile duct [CBD] stones = 248 patients and hepatolithiasis = 13 patients) were included in the study. Percutaneous transhepatic cholangiography was performed, and stones were identified. Percutaneous transhepatic balloon dilation of the papilla of Vater was performed. Then stones were pushed out into the duodenum with a Fogarty balloon catheter. If the stone diameter was larger than 15 mm, then basket lithotripsy was performed before balloon dilation. RESULTS: Overall success rate was 95.7%. The procedure was successful in 97.5% of patients with CBD stones and in 61.5% of patients with hepatolithiasis. A total of 18 major complications (6.8%), including cholangitis (n = 7), subcapsular biloma (n = 4), subcapsular hematoma (n = 1), subcapsular abscess (n = 1), bile peritonitis (n = 1), duodenal perforation (n = 1), CBD perforation (n = 1), gastroduodenal artery pseudoaneurysm (n = 1), and right hepatic artery transection (n = 1), were observed after the procedure. There was no mortality. CONCLUSION: Our experience suggests that percutaneous transhepatic stone expulsion into the duodenum through the papilla is an effective and safe approach in the nonoperative management of the bile duct stones. It is a feasible alternative to surgery when endoscopic extraction fails or is rejected by the patient. PMID- 21638146 TI - Interlocking detachable coil embolization by technique of dumbbell-shaped framing and filling and bridge formation under balloon dilatation for huge conglomerate pulmonary arteriovenous malformation. AB - A 60-year-old woman presented with a conglomerate pulmonary arteriovenous malformation (PAVM) size 8.2 * 7.2 cm on chest X-ray. Feeding arteries were A(a)(7) and A(b)(7) , A(8), and A(10). The diameter and length of the A(b)(7) neck were 15.5 and 16 mm, respectively. After percutaneous transcatheter embolization of A(8) and A(10), PTE of A(a)(7) and A(b)(7) was conducted under balloon occlusion with interlocking detachable coils using a technique of dumbbell-shaped framing and filling, making a bridge from A(b)(7) to the trunk of A(9) and A(10)across A(7). Follow-up computed tomography 10 months after treatment showed marked shrinkage of the PAVM. PMID- 21638147 TI - Efficacy and safety of two different n-butyl-2-cyanoacrylates for the embolization of varicoceles: a prospective, randomized, blinded study. AB - PURPOSE: This was a prospective, randomized, blinded comparative study of the efficacy and safety of two different n-butyl-2-cyanoacrylates (NBCAs) for embolization of varicoceles. METHODS: A total of 112 insufficient spermatic veins (left-sided, n=84; right-sided, n=28) that were diagnosed in 83 adult males were prospectively randomized for blinded embolization with NBCA (n=54; Histoacryl, Braun, Germany) or NBCA-MS (n=58; Glubran2, General Enterprise Marketing, Viareggio, Lucca, Italy). Handling, embolic efficacy, and safety of both NBCAs were compared according the fulfillment of a standardized embolization plan, the occlusive effect on the spermatic vein, and the sticking to the microcatheter. Statistical analysis was performed with the Mann-Whitney U test and the Fisher's exact test. RESULTS: Patients of both study arms were comparable for age and clinical indication. Spermatic vein characteristics were comparable for varicocele classification and embolization side. Both NBCAs were equally efficient in occluding the spermatic vein and blocking reflux (NBCA, n=54/54, 100% vs. NBCA-MS, n=54/57, 94.7%; P=0.244). The embolization plan could be accomplished in an equal number of veins for both groups (NBCA, n=45/54, 83.3% vs. NBCA-MS, n=41/58, 70.7%; P=0.124). Adhesiveness of the glue to the microcatheter was the same in both NBCA groups (NBCA, n=25/54, 46.3% vs. NBCA-MS, n=29/58, 50%; P=0.71). No glue-related complications were noted. CONCLUSIONS: NBCA and NBCA-MS are equally efficient and safe glues for embolization of varicoceles. PMID- 21638148 TI - Early migration and ureteric obstruction of an embolisation coil used to treat massive haemorrhage following percutaneous nephrolithotomy. AB - Renal artery embolization is the treatment of choice for massive hemorrhage after percutaneous nephrolithotomy. Late coil migration is a known complication after embolization. We report an unusual case of early coil migration causing ureteric obstruction and postulate on risk factors for this phenomenon. PMID- 21638149 TI - Examination of the pharmacokinetics of active ingredients of ginger in humans. AB - Ginger extracts have been studied in various clinical trials for different indications. However, the pharmacokinetics of the ginger active constituents in human biological matrices is not well investigated. This study aims to develop a LC-MS/MS method for simultaneous measurement of 6-, 8-, and 10-gingerols and 6 shogaol and study their pharmacokinetics in human plasma and colon tissues. A sensitive LC-MS/MS method was established and validated with a low limit of quantification of 2-5 ng/mL. The intra- and inter-day accuracy ranged from -7.3% to 10.4% and from -9.4% to 9.8%, respectively. The intra- and inter-day precision ranged from 0.9% to 10.9% and from 2.0% to 12.4%, respectively. The glucuronide and sulfate metabolites of 6-, 8-, and 10-gingerols and 6-shogaol in plasma and colon tissues were quantified after hydrolysis with beta-glucuronidase and sulfatase. After oral dosing of 2.0 g ginger extracts in human, free 10-gingerol and 6-shogaol were detected in plasma with peak concentrations (9.5 +/- 2.2 and 13.6 +/- 6.9 ng/mL, respectively) at 1 h after oral administration, but no free 6 gingerol and 8-gingerol were detected in plasma from 0.25 to 24 h. The peak concentrations of glucuronide metabolites of 6-, 8-, and 10-gingerols and 6 shogaol were 0.47 +/- 0.31, 0.17 +/- 0.14, 0.37 +/- 0.19, and 0.73 +/- 0.54 MUg/mL at 1 h, respectively. The peak concentrations of the sulfate metabolites of 6-, 8-, and 10-gingerols and 6-shogaol were 0.28 +/- 0.15, 0.027 +/- 0.018, 0.018 +/- 0.006, and 0.047 +/- 0.035 MUg/mL at 1 h, respectively. Very low concentrations (2-3 ng/mL) of 10-gingerol glucuronide and sulfate were found in colon tissues. Pharmacokinetic analysis showed that half-lives of these four analytes and their metabolites were 1-3 h in human plasma. No accumulation was observed for 6-, 8-, and 10-gingerols and 6-shogaol and their metabolites in both plasma and colon tissues after multiple daily dosing. PMID- 21638150 TI - Ninth annual Mario S. Verani memorial lecture : testing our tests: the evidence bar is rising. Regulatory considerations in the development of cardiac imaging agents. PMID- 21638151 TI - Radiation exposure from radionuclide myocardial perfusion imaging: concerns and solutions. PMID- 21638152 TI - Radiation exposure in diagnostic imaging-use, misuse, or abuse? Part I: the background and science of medical radiation. PMID- 21638153 TI - The future of SPECT MPI: time and dose reduction. PMID- 21638154 TI - Comparison of high efficiency CZT SPECT MPI to coronary angiography. AB - BACKGROUND: The recently introduced cadmium zinc telluride (CZT) SPECT cameras have the potential to reduce radiation exposure to patients and shorten imaging time. So far, there has been only one small study comparing the results of high efficiency CZT SPECT myocardial perfusion imaging (MPI) to invasive coronary angiography. METHODS: All patients who had either a Tc-99m sestamibi or Tl-201 SPECT MPI study using a CZT camera (GE Discovery NM 530c) over a 1-year period followed by a coronary angiogram within 2 months were included. Only patients with a history of CABG surgery were excluded. Standard stress protocols were employed. Rest images were acquired for 5 min and stress supine and prone images for 3 min each. Both MPI studies and coronary angiograms were interpreted by blinded readers. A standard 17-segment model was employed for MPI interpretation, and coronary angiograms were interpreted for the presence of obstructive epicardial coronary artery disease (CAD) defined as >=70% luminal narrowing. Correlation was based on the ability to diagnose obstructive epicardial CAD. RESULTS: Of the 3,111 patients who underwent SPECT imaging using the CZT camera during this time period, 230 patients qualified for the correlation study (mean age 64.2 +/- 11.0 years old, 69% male, and 49% had a history of intracoronary stenting). Tc-99m was used in 76% vs Tl-201 in 24% of the studies. Exercise stress was performed in 60% of patients and vasodilator pharmacologic stress in 40%. Sensitivity was 95%, normalcy rate was 97%, and accuracy was 69% for detecting obstructive CAD. CONCLUSIONS: In this so far largest correlation study between coronary angiography and high efficiency CZT SPECT imaging, a high sensitivity and accuracy for detecting obstructive epicardial CAD was found for this new SPECT camera technology. PMID- 21638155 TI - Evaluation of microalbuminuria in obese children and its relation to metabolic syndrome. AB - Several epidemiologic studies have clearly demonstrated that obesity increases the risk of kidney diseases. We have attempted to evaluate the association of obesity with albuminuria, an early marker of kidney disease, among obese children and its relation to metabolic syndrome. This study included 150 obese children. Blood pressure, fasting blood glucose, plasma insulin and the lipid profile were assessed. The homeostasis model assessment of insulin resistance (HOMA-IR) was used to calculate in vivo insulin resistance. Urinary albumin and creatinine were estimated. Microalbuminuria was detected in 22 (14.7%) of the obese children. Waist circumference, blood pressure, triglyceride, low-density lipoprotein (LDL), insulin resistance and fasting blood glucose were significantly higher in obese children with microalbuminuria than in those with normoalbuminuria and showed significant positive correlations with microalbuminuria. High-density lipoprotein (HDL) was significantly lower in obese children with microalbuminuria than in those with normoalbuminuria, with a significant negative correlation with microalbuminuria. We found that body mass index, abdominal obesity, hypertension, impaired fasting glucose level and insulin resistance significantly increased the odds of microalbuminuria in the obese children enrolled in this study. Moreover, high triglyceride, high LDL and low HDL were significantly associated with microalbuminuria. In our patient group, childhood obesity was a risk factor for the development of microalbuminuria, which in turn was significantly associated with metabolic syndrome and its different constituents. PMID- 21638156 TI - Acute tubulointerstitial nephritis. AB - Acute tubulointerstitial nephritis (TIN) is a frequent cause of acute renal failure, characterised by the presence of inflammatory cell infiltrate in the interstitium of the kidney. Immuno-allergic reaction to certain medications, mainly non-steroidal anti-inflammatory drugs and antibiotics are by far the most important etiology for TIN today, but other situations such as infections, toxins, and vasculitis are known to induce TIN. Incidence of TIN is increasing, probably due to prescription habits and NSAID overuse, representing 3-7% of acute kidney injury in biopsies in children. Avoidance of the causal substance and rapid steroid therapy are hallmarks for patient care, but spontaneous initial recovery is very frequent and the general prognosis seems satisfactory. However, development of chronic TIN, without response to steroid or other immunosuppressive treatment, is possible. As the largest part of TIN is secondary to certain drugs, clear indications in particular for NSAID or antibiotics should be respected to reduce the number of TIN cases. PMID- 21638157 TI - Reducing elevated heart rate in patients with multiple organ dysfunction syndrome by the I (f) (funny channel current) inhibitor ivabradine : MODI (f)Y trial. AB - BACKGROUND: Heart rate (HR) is of relevant prognostic value not only in the general population and patients with cardiovascular disease, but also in critically ill patients with multiple organ dysfunction syndrome (MODS). An elevated HR in MODS patients is associated with a worse prognosis. Beta-blocker (BB) administration has been shown to reduce mortality in MODS. In most cases, negative inotropic effects prevent administration of BBs in MODS patients. In this trial we investigate, whether the "funny current" (I (f)) channel inhibitor ivabradine is able and apt to reduce pathologically elevated HR in MODS patients. We hypothesize that critically ill patients could derive particular benefit from the specific HR-lowering agent ivabradine. METHODS: MODI (f)Y is a prospective, single centre, open label, randomized, controlled two arms, phase II-trial to evaluate the potential of ivabradine to reduce an elevated HR in MODS patients. The primary end point is the proportion of patients with a reduction of HR by at least 10 beats per minute (bpm) within 4 days. This trial will randomize 70 patients (men and women, aged >=18 years) with newly diagnosed MODS, with an elevated HR (sinus rhythm with HR >=90 bpm) and contraindications to BB therapy. Treatment period will last for 4 days. All patients will be followed for 6 months. RESULTS: The first patient was randomized on May 21, 2010. CONCLUSIONS: The MODI (f)Y trial is the first application of ivabradine as a pure heart rate reducing agent in MODS patients. PMID- 21638158 TI - Synthesis and characterization of an unsymmetrical cobalt(III) active site analogue of nitrile hydratase. AB - The design, synthesis, and characterization of an unsymmetrical diamidato-dithiol ligand (H(4) 1, where the hydrogen atoms represent deprotonatable amide and thiol protons) and its cobalt(III) complex, a synthetic analogue of the cobalt containing nitrile hydratase enzyme family, are reported. The ligand was prepared in 24% yield from an overall eight-step synthetic pathway following a modified protocol established in our laboratory that includes two peptide couples using O (1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate as the coupling agent. The ligand and all precursors were characterized by NMR spectroscopy and elemental analysis. The cobalt nitrile hydratase synthetic analogue complex [NBu(4)][Co(1)] was prepared on deprotonating ligand H(4) 1 to [1](4-) on addition of 5 equiv of NaH in N,N-dimethylformamide and adding 1 equiv of CoCl(2) at -40 degrees C under a N(2) atmosphere followed by oxidizing the complex by stirring it overnight open to dry air. The complex [NBu(4)][Co(1)] was isolated after counterion exchange with 1 equiv of NBu(4)Cl followed by crystallization from MeCN/Et(2)O in 71% yield. The structure of the complex was confirmed by X-ray diffraction analysis. Cyclic voltammetry studies on [NBu(4)][Co(1)] in a 0.1 M [NBu(4)][PF(6)]/MeCN solution showed a quasi reversible reduction potential at -1.1 V (vs. Ag/AgCl), and magnetic susceptibility investigations indicated the complex is paramagnetic in both the solid and the solution states as determined from inverse-Gouy and Evans NMR methods, respectively. PMID- 21638159 TI - HIV risk reduction intervention among medically circumcised young men in South Africa: a randomized controlled trial. AB - BACKGROUND: Medical male circumcision (MMC) is now considered one of the best available evidence-based biomedical HIV prevention interventions. However, there is some concern about risks for behavioural disinhibition, or risk compensation, following MMC. PURPOSE: The aim of this study was to test a brief one-session (180 min) group culturally tailored HIV risk reduction counselling intervention among men undergoing medical circumcision in South Africa in order to limit behavioural disinhibition. METHODS: A randomized controlled trial design was employed using a sample of 150 men, 75 in the experimental group and 75 in the control group. Comparisons between baseline and 3-month follow-up assessments on several key behavioural outcomes addressed by the intervention were done. RESULTS: Our study found that behavioural intentions and risk reduction skills significantly increased and sexual risk behaviour (reduction of the number of sexual partners and the number of unprotected vaginal sexual intercourse occasions) significantly decreased in the experimental compared to the control condition. However, male role norms did not change among the intervention conditions over time, while AIDS-related stigma beliefs significantly reduced in both conditions over time. CONCLUSION: Study findings show that a relatively brief (one session) and focused HIV risk reduction counselling can have at least short-term effects on reducing sexual risk behaviours in populations at high risk for behavioural disinhibition following medical male circumcision. PMID- 21638160 TI - A novel method for cryopreservation of individual human spermatozoa. AB - The purpose of this study is to develop a novel method for the cryopreservation and efficient post-thaw recovery of individual or small numbers of human spermatozoa. Spermatozoa equilibrated in cryoprotectant buffer were injected with an intracytoplasmic sperm injection (ICSI) needle into a droplet of cryoprotectant on a homemade cryoleaf. The droplet was of cryoprotectant and seminal plasma at a ratio of 1:1. The sperm-loaded cryoleaf was slowly lowered over and stored in liquid nitrogen. Spermatozoa were thawed in a 37 degrees C oil bath without dilution and centrifugation. To test the fertilizing ability of these spermatozoa, the recovered spermatozoa were injected by ICSI into 1-d-old or in vitro-matured human oocytes. Fresh spermatozoa from the same semen samples served as controls. The trials were performed in two separate experiments. In the first set of experiments, 92 spermatozoa were thawed and carefully investigated. The spermatozoa from percutaneous epididymal sperm aspiration had a motility recovery of 92.9% (13/14); ejaculated spermatozoa had a motility recovery of 61.5% (48/78), and only 1.3% (1/78) was lost. Together in the first and second set of experiments, the fertilization rates for the fresh and frozen-thawed spermatozoa were 67.6% (25/37) and 60.6% (40/66), respectively (P = 0.052). The mean embryo cleavage rates in the fresh and frozen-thawed groups were 88% (22/25) and 85% (34/40), respectively (P = 0.990). This cryopreservation method for individual or small numbers of human spermatozoa was efficient and simple. These findings make this method a promising technique for the clinical application of ejaculated sperm from oligozoospermic patients. PMID- 21638161 TI - Comparison of long-term retinoic acid-based neural induction methods of bone marrow human mesenchymal stem cells. AB - The differentiation of human mesenchymal stem cells (hMSCs) into neural cells in vitro provides a potential tool to be utilized for cell therapy of neurodegenerative disorders. Although previous studies repeated different protocols for the induction of neural cells from hMSCs in vitro, the results were not in complete agreement. In this study, we have attempted to compare three of these neural induction methods; retinoic acid (RA) treatment, RA treatment in serum reduced conditions, and treatment using other chemical compounds (dimethyl sulfoxide and potassium chloride) along with RA by real-time cell analysis and immunofluorescent staining of neural markers. RA treatment led to a slow progression of cells into neural-like morphology with the expression of neural protein neurofilament whereas reducing serum during RA treatment caused a much more extended differentiation process. Additionally, neural-like morphology was persistent in the later periods of differentiation in RA treatment. On the other hand, chemical induction caused cell shrinkages mimicking neural-like morphology in a short time and loss of this morphology along with increased cell death in later periods. Among the three methods compared, RA treatment was the most reliable one in terms of stability of differentiation and neural protein expressions. PMID- 21638162 TI - Polyacrylamide gel for facial wasting rehabilitation: how many milliliters per session? AB - BACKGROUND: Facial lipoatrophy is most distressing for HIV patients in pharmacologic treatment. Nonabsorbable fillers are widely used to restore facial features in these patients. We evaluated the safety and aesthetic outcomes of two samples of HIV+ patients affected by facial wasting who received different filling protocols of the nonabsorbable filler Aquamid(r) to restore facial wasting. METHODS: Thirty-one HIV+ patients affected by facial wasting received injections of the nonabsorbable filler Aquamid for facial wasting rehabilitation. Patients were randomly divided into two groups: A and B. In group A, the facial defect was corrected by injecting up to 8 ml of product in the first session; patients were retreated after every 8th week with touch-up procedures until full correction was observed. In group B, facial defects were corrected by injecting 2 ml of product per session; patients were retreated after every 8th week until full correction was observed. RESULTS: Patients of group A noted a great improvement after the first filling procedure. Patients in group B noted improvement of their face after four filling procedures on average. Local infection, foreign-body reaction, and migration of the product were not observed in either group during follow-up. CONCLUSION: The rehabilitation obtained with a megafilling session and further touch-up procedures and that with a gradual build up of the localized soft-tissue loss seem not to have differences in terms of safety for the patients. However, with a megafilling session satisfaction is achieved earlier and it is possible to reduce hospital costs in terms of gauze, gloves, and other items. PMID- 21638163 TI - Breast sensitivity before and after the L short-scar mammaplasty. AB - BACKGROUND: Breast sensitivity preservation is among the aims of modern breast surgery. Large-volume resection, extensive undermining, and resections at the breast base have been associated with breast sensitivity alterations. The L short scar mammaplasty technique is designed to preserve breast sensitivity by resection of tissue in the middle and inferior portions of the breast, but specifically dissection and preservation of the breast lateral neurovascular pedicle. Using this technique, a prospective study was designed for measurement of breast sensitivity quantitatively and subjectively to determine whether different resection volumes of the breast correlate with alteration of sensitivity postoperatively. METHODS: This study compared the sensitivity of 125 breasts of 64 consecutive patients who underwent mastopexy or breast reduction with the L short-scar mammaplasty technique. Nine points were tested with Semmes Weinstein monofilaments: the nipple and cardinal points on the areola and skin. The tests were performed by the same examiner 1 day before surgery and then 6 and 12 months after surgery. The breasts were divided into three groups according to the resected volume as follows: group A (<=200 g), group B (201-400 g), and group C (>400 g). The patients also were asked to complete written surveys 12 months postoperatively, and the response rate was 100%. RESULTS: Before surgery, larger breasts were observed to have less sensitivity in the areola (groups B and C, P<0.001) and skin (group C, P<0.001). After 12 months, no correlation was found between larger resected volumes and less breast sensitivity. The sensitivity of the nipple-areola complex in all the groups reached levels equal to the preoperative measurement within 12 months after surgery. After 6 and 12 months, significant improvement in skin breast sensitivity with resections greater than 200 g (group B, P=0.002; group C, P<0.001) was observed. After 12 months, 66.4% of the patients and 94.4% of the group C patients (average resection, 499 g) reported equal or better nipple-areolar sensitivity compared with preoperative sensibility. No patient reported areas of areola or nipple insensitivity. CONCLUSION: After the L short-scar mammaplasty, breast sensitivity to pressure in most cases returns to preoperative levels or improves primarily in those with larger breasts. The data of this prospective study contradict the misconception that mammaplasty techniques with resections at the breast base leave the patient with reduced breast sensitivity because the fundamental neural pathways, such as the lateral neurovascular pedicle, are preserved. PMID- 21638164 TI - A clinical study of late seroma in breast implantation surgery. AB - BACKGROUND: The use of mammary implants may lead to a variety of early and delayed complications. The most common delayed complications are capsular contracture and implant failure. Late seroma has seldom been reported. In a long term prospective observational study, cases of late seroma were assessed and recorded. METHODS: Between March 2005 and November 2009, suspected cases of late seroma in patients who had undergone breast augmentation or reconstruction with textured implants were evaluated clinically and with instrumental analyses. An ultrasound-guided needle aspiration of the fluid was performed. RESULTS: In this study, 13 cases of late seroma (8 augmentations and 5 reconstructions) were observed. The overall incidence of this complication was 1.68%. Chemical analyses confirmed the diagnosis of seroma, which recurred in all the patients within days of evacuation. For 12 patients, a reoperation was performed, during which the implant was removed, a total capsulectomy was performed, and in cases of subglandular breast augmentation, the implant pocket was changed to a submuscular placement before a new prosthesis was inserted. One patient underwent a second ultrasound-guided needle aspiration. No seroma recurrence was observed in any of the patients during the follow-up period. CONCLUSION: In case of unexpected breast enlargement after prosthesis implantation, the physician should first rule out infection, then investigate possible friction irritation from either a fold in the device or rubbing of a textured implant. According to the authors' experience and another report, this complication occurs in 1% to 2% of cases. The inclusion of this complication in the informed consent form should be considered. Long-term studies designed to investigate the underlying causes of late seroma are warranted for the prevention and treatment of this complication. PMID- 21638165 TI - Minimally invasive otoplasty: technical details and long-term results. AB - BACKGROUND: Aesthetic correction of the ears requires thorough diagnosis and subsequent treatment of the affected areas. The authors' technique addresses every deformity that might need correction through separate incisions, minimal scars, and conservative dissection without the use of permanent sutures. Correction of the eventual discrepancy between cartilage height and the skin envelope is particularly observed and addressed by means of scaphal reduction. The technical details, long-term results, and complications are evaluated in this article. METHODS: A retrospective review of 100 otoplasties performed between 2005 and 2009 by a single surgeon was conducted. Charts were reviewed and data were collected on the type of deformity corrected, the complications, and the outcome. RESULTS: Minimally invasive otoplasty was performed for 100 patients, primarily women (n=71), with an average age of 23.2 years. In 96 cases, bilateral correction was performed using a combination of concho-mastoid sutures (81%), scapha reduction (76%), antihelical folding (70%), conchal reduction (53%), or earlobe correction (19%). Three patients experienced partial recurrence and needed further correction. Partial upper pole necrosis developed in one patient after undue tension was applied to his dressing, and further correction was needed. Three patients experienced mild partial skin discoloration and epidermolysis that lasted no more than 10 days. Most of the patients were very satisfied (84%) or satisfied (15%) with the procedure after an average follow-up period of 12.4 months. CONCLUSIONS: Restoration of facial harmony through aesthetic correction of the ears is a highly satisfying procedure for both patients and surgeons. Since 1988, the author's unit has pursued and improved a technique that could provide shorter recovery times, minimal morbidity, and predictable results for a population consisting primarily of adults. This minimally invasive technique can achieve these goals with excellent long-term aesthetic outcomes. PMID- 21638166 TI - Diced cartilage versus solid grafts in rhinoplasty. PMID- 21638167 TI - Intraoperative assessment of the umbilicopubic distance: a reliable anatomic landmark for transposition of the umbilicus. AB - BACKGROUND: A clear understanding of the anatomic location and the aesthetic traits of the umbilicus is essential for the plastic surgeon repositioning the umbilicus during an abdominoplasty. Currently no consensus exists regarding the ideal location for this unique aesthetic unit of the abdomen. To their interest, the authors noted that the intraoperative distance from the pubic symphysis to midumbilical stalk measured 15 cm for several consecutive patients. They believe the umbilicopubic distance is another clinically useful and reliable anatomic landmark for the plastic surgeon relocating the umbilicus during an abdominoplasty. METHODS: A retrospective chart review analysis was performed for 40 consecutive patients who underwent abdominoplasty or panniculectomy between July 2009 and May 2010 at the authors' institution. The intraoperative measurement of the umbilicopubic distance (pubic symphysis to midumbilical stalk) was available for 32 of these patients. The average umbilicopubic distance was calculated. Two separate graphs were generated to evaluate the relationship of the umbilicopubic distance to the patients' height and body mass index (BMI). The data were saved and analyzed using Microsoft Office Excel. RESULTS: In the study population, the average intraoperative umbilicopubic distance was found to be 15.05 cm. The results validate the mean umbilicopubic distance of 15.04 cm reported by Dubou and colleagues in 1978. For patients whose stature fell between 145 and 178 cm, the umbilicopubic distance was consistently 15 cm. A tendency toward a higher umbilicus was noted as the patients became taller. The BMI did not seem to influence the location of the umbilicus as measured intraoperatively. CONCLUSION: Translocation of the umbilicus to 15 cm from the pubic symphysis in patients with a stature of 145-178 cm is another clinically useful, safe, and expeditious method for relocation of the umbilicus during an abdominoplasty. PMID- 21638168 TI - Transvaginal natural orifice transluminal endoscopic surgery (NOTES)-assisted laparoscopic adrenalectomy: first clinical experience. AB - BACKGROUND: This study aimed to describe the initial clinical experience of transvaginal natural orifice transluminal endoscopic surgery (NOTES)-assisted laparoscopic adrenalectomy (TNLA) and to evaluate its feasibility and efficacy. METHODS: Between May and December 2010, 11 consecutive women were subjected to TNLA for adrenal tumors at the authors' center. With the patient under general anesthesia, a 5-mm trocar and a 10-mm trocar were inserted in the umbilical edge for conventional operating apparatus, and a 10-mm trocar was inserted in the posterior vaginal fornix for a conventional 30o laparoscope. Dissection was performed according to the method of a standard laparoscopic adrenalectomy. The adrenal gland, its tumor, or both were put into a homemade bag and then removed via the incision of the posterior vaginal fornix after complete resection. RESULTS: A total of 11 TNLAs were performed in this series. Only one conversion to open surgery was performed for one patient with Cushing disease, who was subjected to a splenectomy synchronously for an injury of the spleen. Except for this patient, no patients required a blood transfusion. The median operative time was 102 min (range 80-310 min), and the median estimated blood loss was 80 ml (range 30-800 ml). The median size of the mass in terms of the largest diameter was 4.7 cm (range 2.2-6.6 cm). There was no difference between pre- and postoperative median Female Sexual Function Index (FSFI) total scores (P = 0.102). All the patients except the one who had conversion to open surgery were very satisfied with the cosmetic result. CONCLUSIONS: The findings show TNLA to be a feasible and effective surgical technique that results in excellent cosmesis. It may be an alternative technique for the treatment of properly selected female patients with adrenal tumor. PMID- 21638171 TI - Synchronous femoral hernias diagnosed during endoscopic inguinal hernia repair. AB - BACKGROUND: During totally extraperitoneal (TEP) endoscopic repair of inguinal hernias, it is possible to see the internal opening of the femoral canal. The aim of our study was to determine the incidence of synchronous femoral hernias found in patients undergoing TEP endoscopic inguinal hernia repair. METHOD: This was a retrospective review of prospectively collected data on 362 consecutive patients who underwent 484 TEP endoscopic inguinal hernia repairs during a 5-year period, May 2005 to May 2010. During surgery, both inguinal and femoral canal orifices were routinely inspected. The presence of unilateral or bilateral inguinal and femoral hernias was recorded and repaired accordingly. RESULTS: There were a total of 362 patients. More males (343, 95%) underwent a TEP hernia repair than females (19, 5%). There were more cases of unilateral (240/362, 66%) than bilateral (122/362, 34%) inguinal hernias. A total of 18 cases of synchronous femoral hernias were found during operation. There was a higher incidence of femoral hernia in females (7/19, 37%) compared to males (11/343, 3%) (P < 0.001). None of the femoral hernias were clinically detectable preoperatively. CONCLUSION: Females undergoing elective inguinal hernia repair are more likely to have a synchronous femoral hernia than males. We suggest that all women presenting with an inguinal hernia also have a formal assessment of the femoral canal. TEP endoscopic inguinal hernia repair is an ideal approach as both inguinal and femoral orifices can be assessed and hernias repaired simultaneously during surgery. PMID- 21638172 TI - Robotic gastric banding in children and adolescents: a comparative study. AB - BACKGROUND: Minimally invasive techniques are becoming the gold standard approach to surgical treatment of obesity in adults. Robotic surgical systems have the potential to advance the use and development of minimally invasive procedures. To date, there is no study of outcomes following robotic-assisted gastric banding (RAGB) compared with laparoscopic adjustable gastric banding (LAGB) in children and adolescents. This study was undertaken to compare a single surgeon's results using the da Vinci surgical system for RAGB with those using LAGB in this group of patients. METHODS: A retrospective data analysis of 25 patients who underwent RAGB compared with 50 patients who underwent LAGB. RESULTS: The 75 patients were 8-21 years of age and included 42 (54.5%) girls. Patients in the two groups had similar baseline characteristics including gender, age, body mass index (BMI), and presence of comorbidities. No significant differences in complication rate, length of stay or percent excess weight loss at 1 year were found between the two groups. Mean operating time was significantly shorter for LAGB (50 min) compared with RAGB (74 min; P < 0.001). CONCLUSIONS: This comparative study between RAGB and LAGB in children and adolescents demonstrates the feasibility and safety of RAGB. However, the procedure requires significantly more operative time than LAGB and fails to provide improved patient outcome. PMID- 21638173 TI - Percutaneous drainage for acute calculous cholecystitis. AB - BACKGROUND: Acute calculous cholecystitis is a frequently encountered problem in surgical practice; laparoscopic cholecystectomy (LC) is the standard treatment. LC for acute cholecystitis can be a more difficult procedure than elective LC for cholelithiasis and is associated with increased operating time, higher conversion rate, and more postoperative complications. In the elderly patient with comorbidity, surgery can result in serious complications and even mortality. Percutaneous drainage (percutaneous cholecystostomy; PC) may be an alternative treatment. There is no hard evidence in current literature regarding the safety, success rate, and specific technique of this procedure, nor is there consensus on the indications. AIM: To evaluate the safety and efficacy of PC in treatment of acute calculous cholecystitis in high-risk surgical patients. METHODS: From January 2009 until May 2010, 101 patients with acute calculous cholecystitis were treated, of whom 27 with PC. Of these 27 patients, comorbidity and American Society of Anesthesiologists (ASA) classification were determined, indication for drainage instead of cholecystectomy was recorded, and procedure-related data were collected. Primary outcomes were overall morbidity, mortality, and recurrent biliary events. Secondary outcomes were time to recovery and need for and difficulty of interval laparoscopic cholecystectomy. RESULTS: The cohort included 15 male and 12 female patients with median age of 83 years (range 69-90 years). Most patients were ASA 3 (n = 18) or ASA 2 (n = 8); one patient was ASA 4. Indication for drainage was age and/or comorbidity in 24 cases and duration of symptoms in 3 cases. Antibiotic treatment was given in all but seven patients. The drain was in situ for a median period of 19 days (range 5-57 days). Relief of symptoms occurred in 26 patients; drain luxation occurred in nine patients, only in two patients with clinical consequences. Overall mortality rate was 14.8% (n = 4) with a procedure-related mortality rate of 3.7%. Median time to full recovery was 8 days. With median follow-up of 8 weeks, four patients underwent interval cholecystectomy. CONCLUSIONS: Percutaneous drainage in acute calculous cholecystitis in high-risk patients seems to be a safe and successful treatment option in patients less eligible for surgery. There are many controversies in the current literature, and evidence-based guidelines for the indication of PC in treatment of acute calculous cholecystitis are needed. PMID- 21638174 TI - Laparoscopic repair of large hiatal hernia: impact on dyspnoea. AB - INTRODUCTION: This study aims to examine the impact of laparoscopic repair of large hiatal hernia on dyspnoea severity, respiratory function and quality of life. METHODS: From 2004 to 2008, 30 consecutive patients with large para oesophageal hernia defined as >50% of stomach in the intra-thoracic cavity and minimum follow-up of 2 years were included in this study. All patients had a formal respiratory function test 1 week prior and 3 months after their laparoscopic hiatal hernia repair. Patients rated symptom severity and completed a quality-of-life questionnaire [Gastrointestinal Quality of Life Index (GIQLI)] pre-operatively, and post-operatively at 3 months, 6 months and yearly thereafter. RESULTS: There was no hospital mortality, and the morbidity rate was 10%. In 26 patients with pre-operative dyspnoea, 22 had complete resolution while the remaining 4 had improvement of dyspnoea severity post-operatively. The mean dyspnoea severity index reduced from 2.4 to 1.3 (P < 0.001). Overall, there was 1%, 3% and 3% post-operative increase in forced expiratory volume in 1 s (FEV1), forced vital capacity (FVC) and diffusing capacity of the lung for carbon monoxide (DLCO) values for the whole group, none of which reached statistical significance. For patients with resolution or improvement of dyspnoea after laparoscopic repair, no significant change of respiratory function parameters was demonstrated. GIQLI score improved from a pre-operative value of 85.7 to 107.9 post-operatively (P < 0.001). CONCLUSIONS: We failed to show a significant change in post-operative respiratory function despite clearly demonstrated improvement of respiratory symptoms. Alternative explanations for reduction of dyspnoea severity should be sought. PMID- 21638175 TI - Experience with the use of fibrin sealant plus polyglycolic acid felt at the cut surface of the liver in laparoscopic hepatectomy. AB - BACKGROUND: To minimize bleeding and biliary leakage after open hepatectomy, fibrin sealing is undertaken at the transaction plane of the liver. Recently, clinicians have begun using polyglycolic acid PGA felt as an absorbable cross linker. However, this method has not been well studied for laparoscopic hepatectomy because available laparoscopic devices are quite limited. This study aimed to investigate the feasibility and efficacy of using fibrin sealant with PGA felt in laparoscopic hepatectomy. METHODS: A retrospective analysis of prospectively collected data from 1997 for laparoscopic hepatectomy was performed. Application of fibrin sealant with PGA felt in laparoscopic hepatectomy was begun in February 2009, and the data collected until November 2009 were used. The differences in the perioperative factors were compared including hematocrit and bilirubin concentration changes in the drainage fluid between the surgical procedures conducted with and without the use of fibrin sealant and PGA felt. RESULTS: Fibrin sealant with PGA felt was used in 18 patients who underwent laparoscopic hepatectomy after February 2009. The data for these patients were compared with those for 22 patients who underwent laparoscopic hepatectomy before the start of fibrin sealant use. The operative procedure and devices differed according to the period. No significant differences in preoperative factors were observed between the groups. The use of fibrin sealant had no influence on the operation time or changes in the leukocyte count or serum C-reactive protein. No cases of postoperative bleeding or biliary leakage occurred in either group, and the time course of the perioperative hematocrit and drain bilirubin concentrations did not differ between the two groups. CONCLUSION: Use of fibrin sealant with PGA felt in laparoscopic hepatectomy appears to be feasible, and the outcomes are not inferior to those in the control group. PMID- 21638176 TI - Impact of posterior retroperitoneoscopic adrenalectomy in a tertiary care center: a paradigm shift. AB - BACKGROUND: Posterior retroperitoneoscopic adrenalectomy has substituted its anterior laparoscopic counterpart as the treatment of choice in the management of adrenal tumors at the authors' institution. The authors present their comparative results between these operative techniques, demonstrating the reasons for this change. METHODS: From May 2008 to September 2010, 30 patients underwent posterior retroperitoneoscopic adrenalectomy. Operative time, complications, hospital stay, postoperative pain, and cost were compared with those of 30 selected laparoscopic control subjects treated from 2005 to 2010. Statistical analysis was based on Chi square, the Mann-Whitney U test, the independent-samples t-test, and the Wilcoxon matched pairs test, as appropriate. RESULTS: The median tumor size was 3.8 cm (range, 1.5-8.0 cm) in the retroperitoneoscopic group and 4.9 cm (range, 2.4-8.0 cm) in the laparoscopic group. The median operative time was similar between the two groups (90.0 min; range, 60-165 min vs. 77.5 min; range, 55-120 min; P = 0.138). It was, however, significantly reduced after the 20th case (97.5 min; range, 80-165 min vs. 70 min; range, 60-110 min; P < 0.001) in the retroperitoneoscopic group. The median visual analog pain scores were significantly lower in the retroperitoneoscopic group on both the first and the third postoperative days, respectively (1; range, 0-1 vs. 4; range, 3-6; P < 0.001 and 0; range, 0-1 vs. 3; range, 2-6; P < 0.001). The median postoperative hospital stay also was shorter in the retroperitoneoscopic group (2 days; range, 2-3 days vs. 4 days; range, 3-6 days; P < 0.001). The cost of the posterior approach was significantly less than that of the laparoscopic technique (P < 0.001). CONCLUSIONS: Posterior retroperitoneoscopic adrenalectomy compared with laparoscopic adrenalectomy was safe, fast, and vastly superior in terms of postoperative pain and hospital stay in this series. Because of the ability to reproduce such excellent operative results, the impressive patient recovery, and the significantly reduced operative cost, the authors suggest that the retroperitoneoscopic approach should become the method of choice in minimally invasive adrenal surgery. PMID- 21638177 TI - Laparoendoscopic single-site totally extraperitoneal adult inguinal hernia repair: initial 100 patients. AB - BACKGROUND: This report aims to describe the authors' initial experience with laparoendoscopic single-site (LESS) totally extraperitoneal (TEP) inguinal hernia repair in 100 patients. METHODS: Patients who underwent an elective LESS TEP inguinal hernia repair between December 2008 and September 2010 in a single center were enrolled prospectively in this study. Patient demographic data, hernia characteristics, and operative and postoperative outcomes were analyzed. An Alexis wound retractor was placed through the 2-cm subumbilical incision as a homemade transumbilical access platform after the preperitoneal space was created by a balloon dissector. Standard procedures of TEP all were finished using conventional straight laparoscopic instruments. RESULTS: Of the 100 patients in this study, 2 underwent conversion to LESS transabdominal preperitoneal (TAPP) repair. The remaining 98 patients received successful LESS TEP inguinal hernia repair by a single surgeon. No patient required open or conventional laparoscopic conversion. However, one patient did experience recurrence. The mean operative time was 64.2 min, and the hospital stay was 1.54 days. One patient with a history of bladder surgery had a minor intraoperative bladder injury. No major postoperative complication occurred, but 11 patients had seroma or hematoma, 2 had epididymitis, 2 had urinary tract infection, 1 had wound dehiscence, 1 had wound infection, and 1 had urinary retention. This single-arm observational study was limited by the absence of a control cohort. CONCLUSIONS: Based on our experience, in the hands of experienced laparoscopic surgeons, LESS TEP repair for adult inguinal hernia using the homemade port as an access platform is feasible and safe and provides acceptable operative outcomes. PMID- 21638178 TI - Enodoscopic band ligation (EBL) is superior to endoscopic clipping for the treatment of colonic diverticular hemorrhage. AB - BACKGROUND: Recently, endoscopic band ligation (EBL) has been used to treat colonic diverticular hemorrhage, but the number of EBL cases treated to date has been limited. This study aimed to evaluate the clinical outcomes of EBL in the treatment of colonic diverticular hemorrhage compared with those of endoclips. METHODS: At St. Luke's International Hospital in Tokyo, 66 patients were treated with EBL or endoclips from January 2004 to October 2010. Early rebleeding was defined as clinical evidence of recurrent bleeding within 30 days after initial treatment. Patients' demographics, rate of early rebleeding, and complications were retrospectively evaluated. RESULTS: Of the 66 patients, 18 were treated with EBL. The initial success rate for hemostasis with EBL was 100% with no complications. Early rebleeding was observed in one patient (6%), for whom eversion of a bleeding diverticulum in the sigmoid colon could not be obtained and early loss of the O-band occurred. However, the patient could be retreated with EBL. On the other hand, complete eversion could be obtained for all 10 patients with right-sided diverticula, and no early rebleeding occurred. Endoclips were used to treat 48 patients. Although the initial success rate for hemostasis was 100% without any complications, the rate of early rebleeding was 33% (16 patients), which was significantly higher than the rate for the EBL treated group (P = 0.018). CONCLUSIONS: According to the findings, EBL should be considered safe, effective, and superior to endoclips for the treatment of colonic diverticular hemorrhage. The EBL procedure should be attempted as the initial therapy especially for the right-sided disease. PMID- 21638179 TI - First human surgery with a novel single-port robotic system: cholecystectomy using the da Vinci Single-Site platform. AB - BACKGROUND: Interest in single-incision laparoscopic techniques continues to grow. A learning curve certainly exists, and although the techniques have improved, technical considerations including external clashes, poor visualization of critical structures, and surgeon fatigue remain. Applications of robotics to these newer techniques are evolving. The da Vinci Single-Site robotic surgery platform is a new semi-rigid robotic operative system designed to work with the Intuitive Surgical da Vinci Si operative system. The authors present the first human experience with this new device used to perform single-incision laparoscopic cholecystectomy. This study aimed to present the first human experience performing cholecystectomy with a novel robotic single-port platform. METHODS: After Cleveland Clinic Institutional Review Board approval, patients with symptomatic biliary disease fitting standard criteria for cholecystectomy were enrolled in the study. A total of 13 consecutive single-port, robotic cholecystectomy operations were performed. Patient demographics and operative results were evaluated. RESULTS: All 13 operations were completed successfully with the Single-Site platform. A critical view of safety as determined by the operative surgeon was obtained in all cases. One patient required placement of an additional extra-umbilical trocar for appropriate visualization secondary to gallbladder necrosis, and this incision also was used for eventual drain placement. For two patients, an intraoperative choloangiogram was performed. The mean operative time was 107 +/- 54 min, and the mean docking time was 15 +/- 9 min. Discharge for 11 patients was within 24 h. No significant complications occurred. CONCLUSION: Cholecystectomy using the da Vinci Single-Site system is feasible. Availability of this new semi-rigid, robotic surgery platform may increase access to the potential advantages of single-site surgery. Additional advantages of single-site surgery may be elucidated with further studies. PMID- 21638180 TI - Single-incision laparoscopic colectomy for malignant disease. AB - BACKGROUND: Laparoscopic colectomy has been shown to confer equivalent disease free and overall survival compared to traditional open colectomy. Patients experience benefit in length of hospital stay as well as diminished narcotic use. Single-incision laparoscopic colectomy (SIL-C) may offer additional benefit compared to traditional laparoscopic colectomy without compromising oncologic principles. METHODS: We retrospectively reviewed records of patients who underwent SIL-C and traditional laparoscopic colectomy (TLC) for potentially malignant and malignant disease performed by a single surgeon. SIL-C consisted of a single-port access device with traditional lateral-to-medial laparoscopic technique. RESULTS: Between January and October 2009, 27 SIL-C procedures were performed. Forty-six TLC patients from the prior year were used as controls. Median age was 70 years and 54% were female, with no differences between the groups. The median body mass index (BMI) was 27 kg/m(2) (range = 18.3-39.9) and 26 kg/m(2) (16.6-71.4) for SIL-C and TLC, respectively. The median lymph node harvest was 15 (range = 3-32) and 17 (0-35) for SIL-C and TLC, respectively. The median operative time was 114 min (range = 59-268) and 135 min (45-314) for SIL-C and TLC, respectively. Five SIL-C required additional ports while six TLC required conversion to open technique. The median length of stay was 3 days (range = 2-17) and 5 days (range = 2-11) for SIL-C and TLC, respectively (p = 0.079). There were five significant postoperative complications in the SIL-C group and 16 in the TLC group, including four postoperative ileus and one leak. There were no postoperative deaths in the SIL-C group and two in the TLC group. CONCLUSIONS: SIL-C can be used safely in selected colon cancer patients with no difference in blood loss, OR time, or lymph node retrieval. SIL-C patients may have a shorter LOS. PMID- 21638181 TI - Efficacy of intraperitoneal local anaesthetic techniques during laparoscopic cholecystectomy. AB - BACKGROUND: Pain following laparoscopic cholecystectomy (LC) is a barrier to early discharge. Some studies have demonstrated that local anaesthetic (LA) washed over the liver and gallbladder decreases postoperative pain. In many patients pain is partially of diaphragmatic origin which may not be treated effectively by topical wash. This study assesses the efficacy of LA injected to the peritoneum of the right hemidiaphragm or topical wash with a control group. METHODS: We performed a double-blind randomized sham controlled trial of 128 consecutive subjects who underwent elective LC. Patients received subcutaneous bupivacaine, a diaphragmatic injection of bupivacaine or sham, and topical wash over the liver/gallbladder with bupivacaine or sham depending upon allocation. The primary outcome was VAS pain scores on the ward. Secondary outcomes included VRS pain scores in theatre recovery, analgesic use, physiological observations, time to eating and ambulation, and successful day-case surgery. RESULTS: Pain scores were significantly lower in both LA groups versus control in theatre recovery but only in the subperitoneal diaphragm injection group when the patients returned to the ward. Subperitoneal diaphragm injection was associated with a reduced time in theatre recovery (p = 0.04). CONCLUSIONS: Intraperitoneal techniques of LA during LC decrease postoperative pain and shorten time in theatre recovery. Injection of LA to the right hemidiaphragm is associated with lower pain scores for a longer period following LC than a previously validated wash technique. PMID- 21638182 TI - Laparoscopic salvage surgery for recurrent and metachronous colorectal cancer: 15 years' experience in a single center. AB - BACKGROUND: Laparoscopic salvage surgery for colorectal cancer is a novel but technically challenging option for surgeons. The aim of this study was to evaluate the feasibility and safety of laparoscopic surgery in patients with recurrent or metachronous colorectal cancer in comparison with an open approach. METHODS: The data used in this study were obtained from databases, the data of which were collected prospectively from January 1996 to February 2010. Data pertaining to patients, operations, and short-term outcomes were analyzed and compared between open and laparoscopic salvage groups. RESULTS: Among the 3,425 patients studied, colorectal cancer recurred in 565 patients (16.5%) and 41 patients had colorectal salvage operations. Twenty-six patients with recurrence underwent open surgery and 15 cases underwent laparoscopic surgery. The short term outcomes of the laparoscopic group were comparable with those of the open surgery group or were partly favorable. The five-year disease-free interval and overall survival of recurrent cancer patients were not significantly different from those of the open patients. Metachronous colorectal cancer occurred in 13 patients (0.38%), 5 of whom had open surgery and 6 had laparoscopic salvage. The only significant difference between the groups was a shorter operating time for the laparoscopic group. Late in the study, four patients in the laparoscopic recurrent group and one patient in the metachronous group were converted to open surgery. CONCLUSIONS: Laparoscopic surgery yielded short-term outcomes that were comparable to those of conventional open surgery, in both recurrent and metachronous colorectal cancer patients. Thus, minimally invasive salvage approaches should be considered as a treatment option for the recurrent and the metachronous colorectal cancer patient. PMID- 21638183 TI - First-phase insulin secretion, insulin sensitivity, ghrelin, GLP-1, and PYY changes 72 h after sleeve gastrectomy in obese diabetic patients: the gastric hypothesis. AB - BACKGROUND: The aim of this study was to evaluate the possible role of sleeve gastrectomy (SG) per se in the reversibility of diabetes. METHODS: Insulin secretion and peripheral insulin sensitivity using the intravenous glucose tolerance test (IVGTT) were assessed in 18 obese type 2 diabetic patients and in 10 nondiabetic obese patients before and 3 days after SG, before any food intake and any weight change occurrence. At the same time, ghrelin, GLP-1, and PYY levels were determined. RESULTS: In diabetic patients who had the disease less than 10.5 years, the first phase of insulin secretion promptly improved after SG. The early insulin area under the curve (AUC) significantly increased at the postoperative IVGTT, indicating an increased glucose-induced insulin secretion. The second phase of insulin secretion (late AUC) significantly decreased after SG in all groups, indicating an improved insulin peripheral sensitivity. In all groups, pre- and postoperatively, intravenous glucose stimulation determined a decrease in ghrelin values and an increase in GLP-1 and PYY values. However, in the group of patients with disease duration >10.5 years, the differences were not significant except for the late insulin AUC. Postoperative basal and intravenous glucose-stimulated ghrelin levels were lower than preoperative levels in all groups of patients. Basal and intravenous stimulated GLP-1 and PYY postoperative values were higher than preoperative levels in all groups. CONCLUSIONS: Restoration of the first phase of insulin secretion and improved insulin sensitivity in diabetic obese patients immediately after SG, before any food passage through the gastrointestinal tract and before any weight loss, seem to be related to ghrelin, GLP-1, and PYY hormonal changes of possible gastric origin and was neither meal- nor weight-change-related. Duration of the disease up to 10.5 years seems to be a major cut off in the pathophysiological changes induced by SG. A "gastric" hypothesis may be put forward to explain the antidiabetes effect of SG. PMID- 21638184 TI - Image inversion and digital mirror-image technology aid laparoscopic surgery task performance in the paradoxical view: a randomized controlled trial. AB - BACKGROUND: As laparoscopic surgical procedures increase in complexity, surgeons may find themselves with the laparoscope opposite to their laparoscopic instruments, thus creating the paradoxical viewpoint. We assessed whether surgical task performance in the paradoxical viewpoint would be improved by digitally altering the image or by changing the camera orientation. METHODS: Sixty-one laparoscopically naive operators performed a Peg Transfer task using a trainer box. In the first "round," naive operators were block-randomized to perform the Peg-Transfer task either in the standard view or the paradoxical view. In the second "round," naive operators were positioned in the paradoxical view and block-randomized to having the monitor image as paradoxical (n = 19) or altered by being digitally flipped (mirror-image) (n = 22) or inverted (n = 20). The task consisted of transferring six plastic objects in 5 min (300 s). Scoring was based on the formula of total time = time to completion (max = 300 s) + penalty time (50 s/peg not transferred). RESULTS: In the first round, average total time to perform the Peg Transfer task using the standard view was 215 +/- 20 s, which was significantly less (P < 0.001) than the 563 +/- 13 s for the paradoxical view. In the second round (with all naive operators in the paradoxical viewpoint), the total time for the paradoxical image, digitally flipped image (mirror-image), and inverted image were 561 +/- 12, 449 +/- 25, and 259 +/- 37 s, respectively. The total time for the inverted image was significantly less than both the paradoxical image and digitally flipped image (P < 0.001). The total time for the digitally flipped image was also less than paradoxical image (P < 0.05). The group with the paradoxical image completed 0.8 +/- 0.2 peg transfers, which was less than both the digitally flipped and inverted-view groups (P < 0.05). CONCLUSIONS: This is the first study to demonstrate that when in the paradoxical viewpoint, altering the image on the video monitor, either by digitally flipping or inverting the image, can improve surgical task performance. PMID- 21638185 TI - Operation time and body mass index are significant risk factors for surgical site infection in laparoscopic sigmoid resection: a multicenter study. AB - BACKGROUND: Surgical site infection (SSI) in patients who underwent colorectal surgery is a common complication associated with increased morbidity and costs. The aim of this study was to assess risk factors for SSI in laparoscopic sigmoid resection for benign disease. METHODS: Using a multicenter database of the Swiss Association of Laparoscopic and Thoracoscopic Surgery, we prospectively identified 4,488 patients who underwent laparoscopic colorectal surgery between 1995 and 2008; of these, 2,571 patients who underwent sigmoid resection for benign disease were included. Uni- and multivariate analyses were used to determine risk factors for SSI. RESULTS: The incidence of SSI was 3.5% (90/2,571). Among SSI patients, incisional superficial infections were found in 71%, incisional deep infections in 22%, and organ-space infections in 7%. Patients' age, underlying disease, and surgeons' experience had no impact on SSI. Multivariate analyses showed that operation time >240 min (odds ratio [OR] 1.7; 95% confidence interval [CI] 1.0-2.8), BMI >= 27 kg/m(2) (OR 2.3 [1.3-4.5]), organ lesions (OR 7.9 [2.0-31.8]), and male gender (OR 2.3 [1.2-4.5]) were significant risk factors for SSI. Reoperations in the SSI group were significantly more frequent than in the Non-SSI group (30% vs. 3%; p < 0.001). SSI was associated with a significantly longer median hospital stay (15 days, range = 2-69 vs. 8 days, range = 1-69; p < 0.001) and higher mortality rate (2.2% vs. 0.4%; p = 0.019). CONCLUSION: Significant risk factors for SSI were operation time > 240 min, BMI >= 27 kg/m(2), organ lesions, and male gender. SSI was significantly associated with more reoperations, longer hospital stay, and higher mortality rate. PMID- 21638186 TI - Prospective comparative study of the efficacy of staple-line reinforcement in laparoscopic sleeve gastrectomy. AB - BACKGROUND: Staple-line reinforcement has been used with promising results in laparoscopic gastric bypass in order to reduce leakage, increase staple-line integrity, and diminish staple-site bleeding. The purpose of this study was to determine if staple-line reinforcement with bovine pericardial strips reduces surgical complications of laparoscopic sleeve gastrectomy (LSG). METHODS: This is a prospective comparative study of all patients who underwent LSG by a standard operative team in an 18-month period. Patients were enrolled in group A if they received staple-line reinforcement and in group B when not. The staple line was reinforced with bovine pericardium strips [Peri-Strips Dry (PSD)]. RESULTS: In total, 187 patients, with a median preoperative BMI of 45.3 kg/m(2) (range = 35.1 72.7), underwent LSG. Ninety-six patients were enrolled in group A and 91 in group B; the two groups were comparable in their various characteristics. Morbidity rate representing grade III-IV surgical complications reached 7.4% and mortality rate was 0.5%. Reinforcement with PSD significantly reduced the occurrence of bleeding from the staple line and intra-abdominal collections (P = 0.012 and 0.026). The leak rate was not significantly reduced in group A. Patients in group A required fewer days of hospitalization. CONCLUSIONS: Reinforcement of the staple line in LSG resulted in significantly fewer surgical complications compared to standard stapling of the gastric tube. The additional cost due to the reinforcement of the staple line may be counterbalanced by the reduction in the length of hospitalization. PMID- 21638187 TI - The effects of high-frequency jet ventilation (HFJV) on pneumoperitoneum-induced cardiovascular changes during laparoscopic surgery. AB - BACKGROUND: Standard mechanical ventilation may cause adverse cardiovascular effects in addition to those already related to positive-pressure pneumoperitoneum (PP) during laparoscopic surgery. High-frequency jet ventilation (HFJV) is associated with much less airway pressure, with potentially less influence on venous return, thus potentially it may reduce those effects. The aim of this study was to evaluate the benefits of HFJV to reduce the adverse cardiovascular effects during laparoscopic cholecystectomy. METHODS: We conducted a randomized prospective trial, comparing 12 patients undergoing elective laparoscopic cholecystectomy under conventional mechanical ventilation (control) with 13 similar subjects under HFJV (study). Both groups were categorized as ASA I-II and underwent total intravenous anesthesia. Cardiac functionality was continuously evaluated by analysis of arterial pressure wave changes (Edwards Flo Trac sensor and Vigileo monitor). RESULTS: There was no significant difference between both groups regarding age, gender, BMI, and general medical condition, as well as hemodynamic stability and blood gases throughout surgery. A significant reduction in cardiac output was noted in the control group under PP during the initiation of anti-Trendelenburg position (from 5.6 to 5.0 l/min, P = 0.049). A reciprocal change was observed regarding stroke volume. Such changes were not noticed under HFJV. Total peripheral resistance was significantly increased during PP, and heart rate was not significantly affected throughout surgery in both groups. Unexpectedly, we did not observe marked adverse hemodynamic changes in the control group during PP without position adjustment. CONCLUSIONS: The use of HFJV in upper laparoscopic surgery can impede the adverse cardiovascular changes that usually occur during induction of PP. We also suggest that the use of total intravenous anesthesia (as used in our study) may also lessen the cardiovascular impairment during PP. PMID- 21638188 TI - Laparoscopic implantation of neural electrodes on pelvic nerves: an experimental study on the obturator nerve in a chronic minipig model. AB - BACKGROUND: Laparoscopic exposure of pelvic nerves has opened a new area in the field of neuromodulation. However, electrode design and material deterioration remain issues that limit clinical application. The objective of this study was to evaluate experimentally the laparoscopic implantation of different types of neural electrodes in order to achieve functional and selective electrical stimulation of pelvic nerves. METHODS: This was a prospective comparative study of the laparoscopic implantation and tolerance and efficacy of three monopolar cuff electrodes implanted on the obturator nerve in ten Gottingen minipigs (18-20 months old; 14.5-24 kg body weight). Animals were allocated to two groups. A 3-mm diameter laparoscopic instrument was used during dissection of paravesical fossa and obturator nerve on both sides in order to minimize nerve damage. In all animals, a "split-cylinder" cuff electrode was implanted around the left obturator nerve. On the right side, a "lasso" cuff electrode was implanted in the first group and a "closed-cylinder" cuff was implanted in the second group. Electrical stimulation (0-5 V, 20 Hz) was performed for implanted electrodes on days 0, 7, 15, 30, 45, 60, and 90. Current intensity thresholds were identified by palpation of muscle contraction. Strength developed according to stimulation level and was measured using weight transducers. RESULTS: All procedures were performed by laparoscopy. Mean operative times differed significantly among groups, the shortest being for split-cylinder electrodes (P = 0.0002). No electrical spread phenomena were observed. Initial thresholds were below 1.5 V (range = 0.5-1.3); however, a significant rise was observed, with time to a maximum of 2.7 V (P < 0.0001). Only split-cylinder electrodes remained functional after 3 months. The mean value of maximum strength remained stable during the study period (P = 0.21, NS). CONCLUSIONS: The laparoscopic approach to implanting neuroprostheses seems to be very attractive. Furthermore, this approach could allow highly selective nerve stimulation to be achieved using simpler devices such as split-cylinder monopolar electrodes. PMID- 21638189 TI - Novel technique of overlaying a poly-L: -lactic acid nanosheet for adhesion prophylaxis and fixation of intraperitoneal onlay polypropylene mesh in a rabbit model. AB - BACKGROUND: One problem with polypropylene mesh (PPM) used to repair abdominal wall hernias is dense adhesions to the visceral surface. The authors developed the biocompatible poly-L: -lactic acid (PLLA) nanosheet (thickness < 100 nm), which has the unique ability to adhere tightly to tissues but not to opposing tissues. This study investigated the antiadhesive and fixative characteristics of the PLLA nanosheet after placement of intraperitoneal onlay PPM (IPOM) overlaid with a PLLA nanosheet on intact peritoneum. METHODS: The PLLA nanosheet was fabricated by the spin-coating method and peeling technique with polyvinyl alcohol (PVA) as a supporting film. Two 1.5-cm-square pieces of mesh were implanted on each peritoneal side of the midline incision. The mesh was fixed to the peritoneum with a suture and then overlaid with a 4-cm-square piece of Seprafilm or nanosheet. To examine the fixative property, mesh was overlaid with Seprafilm or nanosheet without a fixed suture. After 4 weeks, mesh adhesion, inflammatory reaction, fixation, and dislocation of mesh were evaluated. RESULTS: Nanosheet-overlaid meshes were flexible and fit over the peritoneum. Adhesion was observed in 10% of the nanosheet-overlaid meshes and in 50% of the Seprafilm overlaid meshes. The adhesion tenacity grade was significantly lower with the nanosheet-overlaid meshes (0.1 +/- 0.1) than with the Seprafilm-overlaid meshes (1.0 +/- 0.4) (p = 0.029), and the percentage of the adhesion area also was lower with the nanosheet-overlaid meshes (1.0 +/- 1.0% vs 8.5 +/- 3.2%; p = 0.037). The mean inflammatory cell counts were lower with the nanosheet-overlaid meshes (p = 0.0023). Regarding the fixative property, 37.5% of the nanosheet-overlaid meshes were fixated on the peritoneum, but no Seprafilm-overlaid mesh was fixated. CONCLUSION: Overlaying of a PLLA nanosheet was effective for adhesion prophylaxis of intraperitoneal mesh. It also may have a possible beneficial effect on fixation of mesh. PMID- 21638190 TI - Multimedia manuscript: inguinal hernia repair by single-incision pediatric endosurgery (SIPES) using the hydrodissection-lasso technique. AB - Many different techniques for laparoscopic inguinal hernia repair have been introduced recently, using either an intraperitoneal [1-3] or an extraperitoneal [4-6] approach. One of the main challenges is to obtain a complete circumferential closure of the sack at the level of the internal ring without injury to the adjacent vas deferens or spermatic vessels. In an effort to separate these structures from the peritoneum before passing a suture around the base of the sack, we developed the hydrodissection-lasso technique, which is performed using a single-incision endosurgical approach.With the patient in Trendelenburg position, an 8-mm skin incision is made in the umbilicus, and a 5 mm trocar is placed in the inferior aspect for the endoscope. A 3-mm Maryland grasper is placed directly through the fascia in the upper part of the incision. Using a 22-gauge needle inserted percutaneously over the internal inguinal ring, saline is injected into the subperitoneal plane circumferentially, hydrodissecting the peritoneum off the vas deferens and vessels and creating a safe space through which the suture can pass without compromising these structures. A 2-mm stab incision is made directly over the internal inguinal ring, and a lasso technique is used to pass two strands of braided polyester suture around the hernia sack, as demonstrated in the video. Both sutures are tied tightly, leaving the knots under the skin. No direct or indirect manipulation of the vas deferens or vessels takes place during any part of the procedure.In contrast to other described techniques [7], the hydrodissection lasso technique can be used for all indirect inguinal hernias in both girls and boys, and hydrodissection itself may be a useful adjunct to any of the other aforementioned techniques. Although an age limit for exclusive high ligation of the hernia sack for indirect inguinal hernias has not been established, the recurrence rate may be higher for adults if the procedure is not combined with inguinal floor reconstruction [8]. At this time, we therefore limit the proposed technique to prepubertal patients.We have performed the described procedure for 22 patients without any recurrences during a maximum follow-up period of 12 months (Table 1). The patients had minimal postoperative pain. There were no complications except for a transient genitofemoral nerve paresis experienced by one girl in whom the hydrodissection was performed using 1% lidocaine instead of the usual normal saline solution. Although the sack was not resected, there were no cases of postoperative hydroceles.To evaluate whether this novel technique is an adequate long-term solution, a prospective clinical trial comparing standard open and single-incision endosurgical inguinal hernia repair using hydrodissection should be performed. PMID- 21638191 TI - Percutaneous endoscopic sigmoid colostomy for irrigation in the management of bowel dysfunction of adults with central neurologic disease. AB - BACKGROUND: Bowel dysfunction results in a major lifestyle disruption for many patients with severe central neurologic disease. Percutaneous endoscopic sigmoid colostomy for irrigation (PESCI) allows antegrade irrigation of the distal large bowel for the management of both incontinence and constipation. This study prospectively assessed the safety and efficacy of PESCI. METHODS: A PESCI tube was placed endoscopically in the sigmoid colon of 25 patients to allow antegrade irrigation. RESULTS: Control of constipation and fecal incontinence was improved for 21 (84%) of the 25 patients. These patients were followed up for 6-83 months (mean, 43 months), with long-term success for 19 (90%) of the patients. No PESCI had to be removed for technical reasons or for PESCI complications. Late removal of the PESCI was necessary for 2 of the 21 patients. A modified St. Marks Fecal Incontinence Score to assess bowel function before and after PESCI showed a highly significant improvement (P < 0.0001). There were no procedure-related deaths. Complications included minor sepsis at the initial PESCI tube site in four patients and bumper migration in two patients, but there were no complications related to the button device. CONCLUSION: This study showed that PESCI is a simple, safe, and effective technique for distal antegrade irrigation in the management bowel dysfunction for selected patients with central neurologic disease. A successful PESCI is very likely to continue functioning satisfactorily for a long time without technical problems or local complications. PMID- 21638192 TI - Open versus endoscopic component separation: a cost comparison. AB - BACKGROUND: The components separation technique (CST) is performed through an open or endoscopic approach. It is unclear whether the costs associated with the endoscopic instruments outweigh any clinical benefit derived from their use and the avoidance of lipocutaneous flaps. This study aimed to compare the direct costs associated with each approach. METHODS: A retrospective review of patients undergoing open or endoscopic CST between 2005 and 2009 was performed. The review compared patient-related variables, length of hospital stay, wound morbidity, and costs associated with the index operation and encounters within a 6-month period. RESULTS: Of the 54 patients identified, 59% underwent endoscopic repair, and 41% had an open CST repair. The patients were similar in age, American Society of Anesthesiology (ASA) score, gender, body mass index (BMI), number of prior surgeries, active abdominal infection, defect size, operating room time, and length of hospital stay. The overall median direct costs were similar between endoscopic and open CST ($9,942 vs. $17,701; p = 0.09). No difference was detected in median operating room costs, but an approximate $7,000 difference was noted between endoscopic and open CST ($1,871 vs. $8,705; p = 0.96). The median mesh costs differed significantly between endoscopic and open CST ($733 vs. $8,415; p = 0.05) as did stapler use costs ($35 vs. $190; p = 0.002). The median cost of endoscopic instruments was $848. Open CST had a 41% major wound morbidity rate compared with 19% in the endoscopic group (p = 0.07). Most of the encounters in the 6-month follow-up period (85%) were related to wound morbidity. The median cumulative direct costs differed between endoscopic and open CST at 3 and 6 months ($12,528 vs. $20,326; p = 0.05). CONCLUSIONS: In a similarly complex group of patients, the total direct costs associated with endoscopic and open CST were similar. Endoscopic instruments made a marginal contribution to the total overall costs, but significant cost contributors were the use of biologic grafts and wound morbidity. PMID- 21638194 TI - A "swing room" model based on regional anesthesia reduces turnover time and increases case throughput. AB - PURPOSE: Our centre recently implemented a new ambulatory care model featuring two alternate regional anesthesia "swing" operating rooms (RA-SRs) managed by a single anesthesiologist. We hypothesized that this model would be associated with decreased turnover times and improved recovery profiles when compared with a traditional model with a single operating room using general anesthesia. METHODS: We conducted a retrospective cohort study of 164 patients scheduled for hand and wrist surgery who were treated in the RA-SRs under brachial plexus blockade, and we compared the findings with a matched historical control group of patients who underwent general anesthesia in a single operating room (GA-OR). The primary endpoint was room turnover time. Secondary endpoints included home discharge time, postoperative interventions for nausea and pain, and number of cases possible per eight-hour day. RESULTS: Patients in the RA-SR group had faster turnover times than patients in the GA-OR group (median [interquartile range]: 15 min [8-22] vs 54 min [49-61], respectively) as well as faster home discharge times (28 min [20-46] vs 156 min [118-215], respectively) (P < 0.0001). In the RA SR group, postoperative antiemetics were used in 3/164 patients (2%) vs 28/164 (17%) in the GA-OR group (P < 0.0001), and opioids were used in 1/164 (0.6%) vs 132/164 (80%), respectively (P < 0.0001). The median number of daily cases possible in the RA-SR group was 56% greater than in the GA-OR group (8.4 [7.5 9.4] vs 5.4 [5.1-5.8], respectively; P < 0.0001). CONCLUSION: Compared with a traditional model using general anesthesia in a single operating room, the implementation of a model using regional anesthesia with two swing operating rooms was associated with reduced room turnover times, improved recovery profiles, and a higher case throughput. PMID- 21638193 TI - Episodic retrieval and the cortical binding of relational activity. AB - Retrieval of episodic memories depends on the successful "re-collection" of event features, such as the time, place, people, thoughts, and feelings associated with a past experience.In neuroimaging studies, ventral regions of the posterior parietal cortex (vPPC) are particularly active when episodic memories are successfully retrieved. A review of the neural correlates of episodic retrieval is presented along with a new theory, cortical binding of relational activity (CoBRA). According to CoBRA, the vPPC acts as a convergence zone that binds episodic features stored in disparate neocortical regions. This process works in conjunction with other known mechanisms, such as those associated with the prefrontal cortex and medial temporal lobe. PMID- 21638195 TI - Obesogenic family types identified through latent profile analysis. AB - BACKGROUND: Obesity may cluster in families due to shared physical and social environments. PURPOSE: This study aims to identify family typologies of obesity risk based on family environments. METHODS: Using 2007-2008 data from 706 parent/youth dyads in Minnesota, we applied latent profile analysis and general linear models to evaluate associations between family typologies and body mass index (BMI) of youth and parents. RESULTS: Three typologies described most families with 18.8% "Unenriched/Obesogenic," 16.9% "Risky Consumer," and 64.3% "Healthy Consumer/Salutogenic." After adjustment for demographic and socioeconomic factors, parent BMI and youth BMI Z-scores were higher in unenriched/obesogenic families (BMI difference = 2.7, p < 0.01 and BMI Z-score difference = 0.51, p < 0.01, respectively) relative to the healthy consumer/salutogenic typology. In contrast, parent BMI and youth BMI Z-scores were similar in the risky consumer families relative to those in healthy consumer/salutogenic type. CONCLUSIONS: We can identify family types differing in obesity risks with implications for public health interventions. PMID- 21638196 TI - Predicting condom use using the Information-Motivation-Behavioral Skills (IMB) model: a multivariate latent growth curve analysis. AB - BACKGROUND: The Information-Motivation-Behavioral Skills (IMB) model often guides sexual risk reduction programs even though no studies have examined covariation in the theory's constructs in a dynamic fashion with longitudinal data. PURPOSE: Using new developments in latent growth modeling, we explore how changes in information, motivation, and behavioral skills over 9 months relate to changes in condom use among STD clinic patients. METHODS: Participants (N = 1281, 50% female, 66% African American) completed measures of IMB constructs at three time points. We used parallel process latent growth modeling to examine associations among intercepts and slopes of IMB constructs. RESULTS: Initial levels of motivation, behavioral skills, and condom use were all positively associated, with behavioral skills partially mediating associations between motivation and condom use. Changes over time in behavioral skills positively related to changes in condom use. CONCLUSIONS: Results support the key role of behavioral skills in sexual risk reduction, suggesting these skills should be targeted in HIV prevention interventions. PMID- 21638197 TI - Media coverage of direct-to-consumer genetic testing. AB - Media coverage of Direct-to-Consumer (DTC) genetic testing shapes public perception of such testing. The purpose of this study was to determine and assess the themes presented by U.S. news media regarding DTC genetic testing. We performed a Lexis-Nexis search with the keywords "Direct-to-Consumer" and "genetic test" for news stories published from 2006-2009. The sample was coded on themes of genetic determinism, privacy, discrimination, validity, regulation, the Genetic Information Nondiscrimination Act (GINA), utility, and cost. Ninety-two news stories were included. Stories displayed moderate genetic determinism and were neutral about validity and utility. Stories indicated that insurance and employers were the most likely sources of discrimination, yet identified the physicians and DTC companies as groups most likely to violate privacy. Stories claimed lack of regulation would harm consumers, but most post-GINA stories did not discuss the law. The costs of tests were frequently included. The results of this study show a broad range of views toward DTC genetic testing and its potential impacts. The genetics community should be aware that the public has been exposed to multiple views of DTC genetic testing when discussing these tests. PMID- 21638198 TI - MicroRNAs 130a/b are regulated by BCR-ABL and downregulate expression of CCN3 in CML. AB - Chronic Myeloid Leukaemia (CML) is a myeloproliferative disorder characterized by the expression of the oncoprotein, Bcr-Abl kinase. CCN3 normally functions as a negative growth regulator, but it is downregulated in CML, the mechanism of which is not known. MicroRNAs (miRNAs) are small non-coding RNAs, which negatively regulate protein translation by binding to the complimentary sequences of the 3' UTR of messenger RNAs. Deregulated miRNA expression has emerged as a hallmark of cancer. In CML, BCR-ABL upregulates oncogenic miRNAs and downregulates tumour suppressor miRNAs favouring leukaemic transformation. We report here that the downregulation of CCN3 in CML is mediated by BCR-ABL dependent miRNAs. Using the CML cell line K562, we profiled miRNAs, which are BCR-ABL dependent by transfecting K562 cells with anti-BCR-ABL siRNA. MiRNA expression levels were quantified using the Taqman Low Density miRNA array platform. From the miRNA target prediction databases we identified miRNAs that could potentially bind to CCN3 mRNA and reduce expression. Of these, miR-130a, miR-130b, miR-148a, miR-212 and miR-425-5p were significantly reduced on BCR-ABL knockdown, with both miR 130a and miR-130b decreasing the most within 24 h of siRNA treatment. Transfection of mature sequences of miR-130a and miR-130b individually into BCR ABL negative HL60 cells resulted in a decrease of both CCN3 mRNA and protein. The reduction in CCN3 was greatest with overexpression of miR-130a whereas miR-130b overexpression resulted only in marginal repression of CCN3. This study shows that miRNAs modulate CCN3 expression. Deregulated miRNA expression initiated by BCR-ABL may be one mechanism of downregulating CCN3 whereby leukaemic cells evade negative growth regulation. PMID- 21638200 TI - [HTLV and "donating" milk]. AB - In France, the screening for human T-cell leukemia/ lymphoma virus type 1 and 2 (HTLV-1 and HTLV-2) during the donation of human milk has been carried out from 1992 with the application of the circular DGS 24 November 1992. The screening for antibodies against these viruses is regulated and done systematically during every donation of milk. Breast feeding being the main mode of transmission of the HTLV-1, the last ministerial decree of 25 August 2010 has made the screening test compulsory for the anonymous donation and for the personalized donation (of a mother for her own child) from all women including those affected by the infection. The milk delivered by milk banks is pasteurized (62.5 degrees C for 30 minutes) before freezing at -18 degrees C, which inactivates the pathogens. This double means of prevention of the transmission of the HTLV-1 paradoxically seems disproportionate in the absence of any precautionary measure in the case of direct breast-feeding and the use of mother's raw milk. Indeed, in most neonatal intensive care units in maternity hospitals, unpasteurized milk is administered to the neonates without any systematic preliminary testing of the serological HTLV-1 status of the mother. An increased sensitization of the community of the obstetricians, midwives and neonatologists by the Association of the Milk Banks of France (ADLF) and the Societe de pathologie exotique could address the issue of screening for HTLV-1 in "donated" milk and breast-feeding. PMID- 21638199 TI - Adjuvant chemotherapy for breast cancer in older women: emerging evidence to aid in decision making. AB - To prevent breast cancer-related recurrence and death, adjuvant therapy, including chemotherapy, is given. The decision to deliver chemotherapy requires careful weighing of the risk of toxicity versus the estimated benefit. The risk and benefit are based on information from clinical trials, statistical models, and past clinical experience . Compared to younger patients, it is perceived that older patients have cancers that are lower risk, gain less benefit from chemotherapy, and are at higher risk of toxicity. There is now strong evidence that healthy older women tolerate treatment and stand to gain the same benefits from treatment as do younger women. Numeric age alone, therefore, does not justify withholding adjuvant chemotherapy. New tools to aid in the decision are needed. Fortunately, the expected great increase in the size of the geriatric population spawned the field of geriatric oncology and the development of brief, practical versions of the Comprehensive Geriatric Assessment (CGA) for use in busy oncology clinics are in sight. It is time for us to incorporate elements of the CGA into practice, to systematically identify older patients at substantial risk of toxicity. For frail older women with breast cancer, no therapy or less toxic therapies can be considered, some of which are suggested herein. In addition, as always in oncology, physicians and patients should look for and participate in clinical trials that will define how to treat cancer, especially in older patients, in the future. PMID- 21638201 TI - [Malaria in Morocco: from pre-elimination to elimination, what risks for the future?]. AB - By basing itself essentially on the data of literature, this work redraw the historic periods of malaria in Morocco since 1912 at this day. The analysis of entomological, historic and current epidemiological situations of malaria in the Maghreb countries allows to identify elements of reflection on the risk in Morocco. PMID- 21638202 TI - Pathway-specific alteration of synaptic plasticity in Tg2576 mice. AB - Various animal models of Alzheimer disease (AD) are characterized by deficits in spatial memory that are causally related to altered synaptic function and impairment of long-term potentiation (LTP) in the hippocampus. In Tg2576 AD mice, we compared LTP in 2 major hippocampal pathways, Schaffer collateral (SC) and mossy fiber (MF) pathways. Whereas LTP was completely abolished in the SC pathway of Tg2576 mice, we found no decrease in LTP induced by stimulation of the MF pathway. In fact, we found that in the MF pathway, LTP was slightly, but significantly, enhanced compared with that in the MF pathway of WT littermates. This pathway-specific impairment of LTP is not attributable to alterations in transmitter release, as indicated by an unaltered paired-pulse ratio. These results suggest that the spatial memory deficits normally seen in AD models arise primarily from LTP impairment at the SC pathway. PMID- 21638203 TI - Identification and functional characterization of ion channels in CD34(+) hematopoietic stem cells from human peripheral blood. AB - Hematopoietic stem cells (HSCs) are used therapeutically for hematological diseases and may also serve as a source for nonhematopoietic tissue engineering in the future. In other cell types, ion channels have been investigated as potential targets for the regulation of proliferation and differentiation. However, the ion channels of HSCs remain elusive. Here, we functionally characterized the ion channels of CD34(+) cells from human peripheral blood. Using fluorescence-activated cell sorting, we confirmed that the CD34(+) cells also express CD45 and CD133. In the CD34(+)/CD45(+)/CD133(high) HSCs, RT-PCR of 58 ion channel mRNAs revealed the coexpression of Kv1.3, Kv7.1, Nav1.7, TASK2, TALK2, TWIK2, TRPC4, TRPC6, TRPM2, TRPM7, and TRPV2. Whole-cell patch clamp recordings identified voltage-gated K(+) currents (putatively Kv1.3), pH sensitive TASK2-like back-ground K(+) currents, ADP-ribose-activated TRPM2 currents, temperature-sensitive TRPV2-like currents, and diacylglycerol-analogue activated TRPC6-like currents. Our results lend new insight into the physiological role of ion channels in HSCs, the specific implications of which require further investigation. PMID- 21638204 TI - Amygdalar connections in the lesser hedgehog tenrec. AB - The present study analyses the overall extrinsic connectivity of the non olfactory amygdala (Ay) in the lesser hedgehog tenrec. The data were obtained from tracer injections into the lateral and intermediate portions of the Ay as well as several non-amygdalar brain regions. Both the solitary and the parabrachial nucleus receive descending projections from the central nucleus of the Ay, but only the parabrachial nucleus appears to project to the Ay. There is one prominent region in the ventromedial hypothalamus connected reciprocally with the medial and central Ay. Amygdalar afferents clearly arise from the dorsomedial thalamus, the subparafascicular nuclei and the medial geniculate complex (GM). Similar to other subprimate species, the latter projections originate in the dorsal and most caudal geniculate portions and terminate in the dorsolateral Ay. Unusual is the presence of amygdalo-projecting cells in the marginal geniculate zone and their virtual absence in the medial GM. As in other species, amygdalo striatal projections mainly originate in the basolateral Ay and terminate predominantly in the ventral striatum. Given the poor differentiation of the tenrec's neocortex, there is a remarkable similarity with regard to the amygdalo cortical connectivity between tenrec and rat, particularly as to prefrontal, limbic and somatosensorimotor areas as well as the rhinal cortex throughout its length. The tenrec's isocortex dorsomedial to the caudal rhinal cortex, on the other hand, may not be connected with the Ay. An absence of such connections is expected for primary auditory and visual fields, but it is unusual for their secondary fields. PMID- 21638205 TI - Age-dependent alteration of TGF-beta signalling in osteoarthritis. AB - Osteoarthritis (OA) is a disease of articular cartilage, with aging as the main risk factor. In OA, changes in chondrocytes lead to the autolytic destruction of cartilage. Transforming growth factor-beta has recently been demonstrated to signal not only via activin receptor-like kinase 5 (ALK5)-induced Smad2/3 phosphorylation, but also via ALK1-induced Smad1/5/8 phosphorylation in articular cartilage. In aging cartilage and experimental OA, the ratio ALK1/ALK5 has been found to be increased, and the expression of ALK1 is correlated with matrix metalloproteinase-13 expression. The age-dependent shift towards Smad1/5/8 signalling might trigger the differentiation of articular chondrocytes with an autolytic phenotype. PMID- 21638206 TI - Effects of oxygen and culture system on in vitro propagation and redifferentiation of osteoarthritic human articular chondrocytes. AB - Regenerative medicine-based approaches for the repair of damaged cartilage rely on the ability to propagate cells while promoting their chondrogenic potential. Thus, conditions for cell expansion should be optimized through careful environmental control. Appropriate oxygen tension and cell expansion substrates and controllable bioreactor systems are probably critical for expansion and subsequent tissue formation during chondrogenic differentiation. We therefore evaluated the effects of oxygen and microcarrier culture on the expansion and subsequent differentiation of human osteoarthritic chondrocytes. Freshly isolated chondrocytes were expanded on tissue culture plastic or CultiSpher-G microcarriers under hypoxic or normoxic conditions (5% or 20% oxygen partial pressure, respectively) followed by cell phenotype analysis with flow cytometry. Cells were redifferentiated in micromass pellet cultures over 4 weeks, under either hypoxia or normoxia. Chondrocytes cultured on tissue culture plastic proliferated faster, expressed higher levels of cell surface markers CD44 and CD105 and demonstrated stronger staining for proteoglycans and collagen type II in pellet cultures compared with microcarrier-cultivated cells. Pellet wet weight, glycosaminoglycan content and expression of chondrogenic genes were significantly increased in cells differentiated under hypoxia. Hypoxia-inducible factor-3alpha mRNA was up-regulated in these cultures in response to low oxygen tension. These data confirm the beneficial influence of reduced oxygen on ex vivo chondrogenesis. However, hypoxia during cell expansion and microcarrier bioreactor culture does not enhance intrinsic chondrogenic potential. Further improvements in cell culture conditions are therefore required before chondrocytes from osteoarthritic and aged patients can become a useful cell source for cartilage regeneration. PMID- 21638207 TI - Primary cilia and organogenesis: is Hedgehog the only sculptor? AB - The primary cilium is a small microtubule-based organelle projecting from the plasma membrane of practically all cells in the mammalian body. In the past 8 years, a flurry of papers has indicated a crucial role of this long-neglected organelle in the development of a wide variety of organs, including derivatives of all three germ layers. A common theme of these studies is the critical dependency of signal transduction of the Hedgehog pathway upon functionally intact cilia to regulate organogenesis. Another common theme is the role that the cilium plays, not necessarily in the determination of the embryonic anlagen of these organs, although this too occurs but rather in the proliferation and morphogenesis of the previously determined organ. We outline the various organ systems that are dependent upon primary cilia for their proper development and we discuss the cilia-dependent roles that Sonic and Indian Hedgehog play in these processes. In addition and most importantly for the field, we discuss the controversial involvement of another major developmental pathway, Wnt signaling, in cilia-dependent organogenesis. PMID- 21638208 TI - Characterization and comparison of telomere length, telomerase and reverse transcriptase activity and gene expression in human mesenchymal stem cells and cancer cells of various origins. AB - We have characterized and compared the telomere length, telomerase, reverse transcriptase (RT) activity and expression of genes implicated in cancer and in pluripotency, in human mesenchymal stem cells (MSCs) derived from dental papilla tissue, umbilical cord matrix and adipose tissue and in cancer cells (MDA-MB-231, U-87 MG, and MCF-7). MRC-5 fetal fibroblasts and adult muscle cells were used as somatic cell controls. Telomere length was significantly (P<0.05) higher in MSCs and somatic cells (7.2-9.3 kb) than in cancer cell lines (3.9-6 kb). However, the relative telomerase activity (RTA) in the cancer cell lines was significantly (P<0.05) higher than that of MSCs and somatic cells. RTA tended to be slightly higher in MSCs but no significant differences were observed between some cancer cells and MSCs. However, RTA was not detected in somatic cells. Although differentially displayed, the expression of genes related to cancer (BCL-2, p53, NF-kappaB, TGF-beta, VEGF) and transcription and pluripotency (OCT4, NANOG, STAT3, REX1) were commonly observed in MSCs and cancer cells. Thus, endogenous non-telomerase RTA might be a potential biological marker or regulator among MSCs and cancer cells. Further, by sharing the biological and molecular markers of self-renewal and proliferation with cancer cells, MSCs might play a contributory role as tissue resident stem cells in tumor development. PMID- 21638209 TI - TGF-beta1 -> SMAD/p53/USF2 -> PAI-1 transcriptional axis in ureteral obstruction induced renal fibrosis. AB - Chronic kidney disease constitutes an increasing medical burden affecting 26 million people in the United States alone. Diabetes, hypertension, ischemia, acute injury, and urological obstruction contribute to renal fibrosis, a common pathological hallmark of chronic kidney disease. Regardless of etiology, elevated TGF-beta1 levels are causatively linked to the activation of profibrotic signaling pathways initiated by angiotensin, glucose, and oxidative stress. Unilateral ureteral obstruction (UUO) is a useful and accessible model to identify mechanisms underlying the progression of renal fibrosis. Plasminogen activator inhibitor-1 (PAI-1), a major effector and downstream target of TGF beta1 in the progression of several clinically important fibrotic disorders, is highly up-regulated in UUO and causatively linked to disease severity. SMAD and non-SMAD pathways (pp60(c-src), epidermal growth factor receptor [EGFR], mitogen activated protein kinase, p53) are required for PAI-1 induction by TGF-beta1. SMAD2/3, pp60(c-src), EGFR, and p53 activation are each increased in the obstructed kidney. This review summarizes the molecular basis and translational significance of TGF-beta1-stimulated PAI-1 expression in the progression of kidney disease induced by ureteral obstruction. Mechanisms discussed here appear to be operative in other renal fibrotic disorders and are relevant to the global issue of tissue fibrosis, regardless of organ site. PMID- 21638210 TI - Efficacy of plant-mediated synthesized silver nanoparticles against hematophagous parasites. AB - The purpose of the present study was to investigate the acaricidal and larvicidal activity against the larvae of Haemaphysalis bispinosa Neumann (Acarina: Ixodidae) and larvae of hematophagous fly Hippobosca maculata Leach (Diptera: Hippoboscidae) and against the fourth-instar larvae of malaria vector, Anopheles stephensi Liston, Japanese encephalitis vector, Culex tritaeniorhynchus Giles (Diptera: Culicidae) of synthesized silver nanoparticles (AgNPs) utilizing aqueous leaf extract from Musa paradisiaca L. (Musaceae). The color of the extract changed to light brown within an hour, and later it changed to dark brown during the 30-min incubation period. AgNPs results were recorded from UV-vis spectrum at 426 nm; Fourier transform infrared (FTIR) analysis confirmed that the bioreduction of Ag(+) ions to silver nanoparticles are due to the reduction by capping material of plant extract, X-ray diffraction (XRD) patterns clearly illustrates that the nanoparticles formed in the present synthesis are crystalline in nature and scanning electron microscopy (SEM) support the biosynthesis and characterization of AgNPs with rod in shape and size of 60-150 nm. After reaction, the XRD pattern of AgNPs showed diffraction peaks at 2theta = 34.37 degrees , 38.01 degrees , 44.17 degrees , 66.34 degrees and 77.29 degrees assigned to the (100), (111), (102), (110) and (120) planes, respectively, of a faced centre cubic (fcc) lattice of silver were obtained. For electron microscopic studies, a 25 MUl sample was sputter-coated on copper stub, and the images of nanoparticles were studied using scanning electron microscopy. The spot EDX analysis showed the complete chemical composition of the synthesized AgNPs. The parasite larvae were exposed to varying concentrations of aqueous extract of M. paradisiaca and synthesized AgNPs for 24 h. In the present study, the percent mortality of aqueous extract of M. paradisiaca were 82, 71, 46, 29, 11 and 78, 66, 38, 31and 16 observed in the concentrations of 50, 40, 30, 20, 10 mg/l for 24 h against the larvae of H. bispinosa and Hip. maculata, respectively. The maximum efficacy was observed in the aqueous extract of M. paradisiaca against the H. bispinosa, Hip. maculata, and the larvae of A. stephensi, C. tritaeniorhynchus with LC(50) values of 28.96, 31.02, 26.32, and 20.10 mg/lm, respectively (r (2) = 0.990, 0.968, 0.974, and 0.979, respectively). The synthesized AgNPs of M. paradisiaca showed the LC(50) and r (2) values against H. bispinosa, (1.87 mg/l; 0.963), Hip. maculata (2.02 mg/l; 0.976), and larvae of A. stephensi (1.39; 0.900 mg/l), against C. tritaeniorhynchus (1.63 mg/l; 0.951), respectively. The chi (2) values were significant at p < 0.05 level. PMID- 21638212 TI - The introduction of mobile telephony is characterized by an unprecedented success. Editorial. PMID- 21638211 TI - KIR gene diversity in Mexican mestizos of San Luis Potosi. AB - Natural killer (NK) cell function is regulated by different types of membrane bound receptors of which killer-cell immunoglobulin-like receptors (KIRs) are the most complex and diverse. KIRs are encoded by 17 different genes located within the leukocyte receptor complex (19q13.4). The frequency with which KIR gene features are present in different human populations differs. Here, we present our results on the KIR gene diversity observed in a large group of mestizos from the central Mexican city of San Luis Potosi. In total, 53 different KIR genotypes were observed, 47 with previously described gene profiles and six harboring novel KIR gene combinations. Group A homozygous haplotypes were seen in 102 individuals (34%), while group B homozygous haplotypes were present in 45 (15%). Heterozygous combinations of groups A and B haplotypes were seen in 153 individuals (51%). Haplotype frequency estimations based on a true content of 600 chromosomes showed a relatively balanced proportion of group A (59.5%) and group B (40.5%) haplotypes in our study population. A homozygous combination of the cA01|tA01 haplotype was present in 33% of the population with other frequent combinations being cA01|tA01, cB03|tB01 in 14.7% and cA01|tA01, cB02|tA01 in 12%. The dendrogram derived from activating KIR gene phylogenetic analysis revealed five clearly distinct clades corresponding to African, East Asian, Arab/Caucasoid, Mexican mestizo/Amerindian and South Asian populations. Our results illustrate the genetic contribution that Caucasoid and Amerindian populations have made toward present-day Mexicans and suggest an important Southeast Asian genetic contribution to native Amerindian populations. PMID- 21638213 TI - Children's health and RF EMF exposure. Views from a risk assessment and risk communication perspective. AB - A review of the scientific literature published until August 2010, covering epidemiological studies on the effects of RF EMF exposure on the incidence of brain tumours and leukaemia in children as well as experimental studies on RF EMF effects on cognition and CNS in children, reveals no or only scant evidence for the assumption that RF EMF exposure poses a hazard to children. This result is at odds with public risk perceptions, i.e. prevailing concerns of (at least part of) the public about adverse health effect of RF EMF. Consequences for risk communication are discussed. A scientifically sound and easy-to-understand risk communication is especially exacerbated by the fact that current risk assessments cannot exclude that RF EMF might have adverse health effects due remaining knowledge gaps, but especially due to the impossibility to prove a non-effect. PMID- 21638214 TI - Assessment of exposure to mobile telecommunication electromagnetic fields. AB - Typical general public exposures around mobile radio service base stations consume only tiny fractions of exposure levels. Maximal immissions at maximal transmit power of base stations amount to several percent of power density reference levels; typical immission levels are about one tenth of a percent or even less. The distance to base stations is no reliable exposure classifier. More important are the orientation relative to the main lobe of the station and sight conditions from measurement point to the base station. Mobile phones cause higher exposures to the user than base stations. At maximal transmit power up to 80 percent of the basic restrictions are consumed. Therefore, actions to minimize exposure to mobile phones, e.g. by using a headset, have a larger potential than shielding against emissions from base stations. Both base stations and mobile phones apply power control mechanisms, capable to significantly reducing the transmit power and the associated exposure depending on the communication traffic. Present research investigates, whether children are more exposed to mobile telecommunication systems than adults. PMID- 21638215 TI - Wireless communication fields and non-specific symptoms of ill health: a literature review. AB - This is an update of a previous systematic review on the association between radiofrequency electromagnetic field (RF-EMF) exposure and health-related quality of life that included studies published before August 2007 [1]. Since then, nine randomized trials addressed short-term exposures from close-to-body RF-EMF sources such as mobile phones, and two observational studies investigated the effects of mobile phone use on health-related quality of life. Six randomized trials addressed short-term far-field exposure arising, for instance, from mobile phone base stations, and eight studies evaluated the effects of environmental far field RF-EMF exposure. In most of the randomized trials, no exposure-response association was observed. The sporadically reported associations did not show a consistent pattern regarding the type of symptoms or the direction of the effects (increase/decrease). Similarly, most of the recent observational studies did not show associations between RF-EMF exposure and non-specific symptoms. However, the exposure gradients were small and possible exposure misclassification is a limitation of these studies. Longitudinal studies as well as studies in children and adolescents are scarce. In summary, recent research did not indicate health related quality of life to be affected by RF-EMF exposure in our everyday environment. Furthermore, none of the studies showed that individuals with self reported electromagnetic hypersensitivity (EHS) were more susceptible to RF-EMF than the rest of the population. Nevertheless, the rapid technological development and anticipated increase in exposure levels warrant the conduct of further longitudinal studies. Due to the widespread use of wireless communication technologies potential adverse health effects would have major public health consequences. PMID- 21638216 TI - Comparative health risk assessment of electromagnetic fields. AB - Comparative health risk assessment of electromagnetic fields (EMFs) has been performed in their entire frequency range from static fields until ionizing radiation. Due to considerable change of physical and biologic interaction and the nature of potential adverse health effects comparison was based on the carcinogenic potential of environmental EMF exposure levels. There was a need for sophisticated discrimination of levels of evidence as well as the available database. Conclusions were based on a synoptic view to results of different scientific approaches such as theoretical and biologic interaction modeling, evidence for accumulative effects, in vitro and in vivo investigations and epidemiologic studies. The comparative assessment revealed significant differences of objective results and public risk perception, and puts EMF risks into perspective. It highlights the necessity for individual's responsible behavior in terms of prudent avoidance. The comparison indicates where risk awareness might merit priority. This is not restricted to the UV range but includes also other exposures such as to nocturnal light or within infrared cabins. PMID- 21638217 TI - [Outpatient cardiac phase III rehabilitation at a Viennese institution - retrospective cohort study]. AB - BACKGROUND: This retrospective cohort study analyses effectiveness and sustainability of the current cardiac Phase III (Ph-III) rehabilitation program, provided by the Centre for Outpatient Rehabilitation (ZAR). METHODS: We analysed routine data of 451 intervention group patients (IG, with Ph-III) and 781 control group patients (KG, without Ph-III). RESULTS: In a median observation period of 2.73 years we found 30% less cases of death in the IG based on the mortality risk observed in the KG (rr = 0.70; p = 0.108). However, we registered more re-events, mainly stent implantations in the IG (rr = 1.34; p = 0.095). Groups differed in some baseline characteristics. CONCLUSIONS: The lower mortality risk by trend might be explained by the close-meshed care, the IG patients' more health conscious behaviour or a selection bias of the KG (e.g. more severe underlying disease). The causality of potential positive effects cannot be confirmed by this study because of the study design. PMID- 21638218 TI - Position of the mental foramen: an anatomical study. AB - Knowledge of the position of the mental foramen is very important during all surgical procedures and it needs to be considered before all surgical procedures in the mandible region. The aim of this study was to determine the position of the mental foramen in the dissected human cadaver specimens. Four hundred hemimandible specimens from human cadavers were dissected and analyzed for the position of the mental foramen. The most common position of the mental foramen investigated - using anatomical dissection - was between the first and second mandibular premolars. These investigations provide relevant data for clinical anatomy, especially when planning oral operative treatment in the premolar area. PMID- 21638221 TI - Blunted accumbal dopamine response to cocaine following chronic social stress in female rats: exploring a link between depression and drug abuse. AB - RATIONALE: Women have twice the risk as men to develop depression. Approximately, 24% of major depression disorder cases have comorbid disorders with substance abuse. Several central systems, including dopaminergic and serotonergic pathways, are thought to be involved in such comorbidity. OBJECTIVES: The present study established a chronic social stress model in female rats, which produces some cardinal features of depressive-like symptoms. Further, we examined the effects of acute cocaine on dopamine (DA) and serotonin (5-HT) in the nucleus accumbens (NAc) using this model. METHODS: Female Long-Evans rats confronted a nursing dam in its home cage for 30 min twice daily for 21 days. The non-stressed control group was handled daily throughout the experiment. During the 21 days of stress, behaviors during confrontations, weight, preference for saccharin, and estrous cycles were measured. Ten days after the last confrontation, the experimental rat was challenged with 10 mg/kg of cocaine, and levels of DA and 5-HT in the NAc were measured using in vivo microdialysis. RESULTS: During the course of daily confrontations for 21 days, the experimental females significantly increased the duration of immobility, reduced weight gain and the preference for saccharin, and disrupted estrous cycles during the stress. Chronic social stress significantly attenuated cocaine-induced DA levels, and to some extent, attenuated a percent change of 5-HT compared to the non-stressed control group. CONCLUSIONS: Chronic social defeat stress for 21 days induced physiological and behavioral depression relevant deficits and blunted response of dopaminergic and to some extent, serotonergic neurons to cocaine challenge in females. PMID- 21638222 TI - Effects of acute administration of nicotine, amphetamine, diazepam, morphine, and ethanol on risky decision-making in rats. AB - RATIONALE: Most individuals can accurately assess the risks and rewards associated with choice alternatives and decide accordingly; however, drug users often display maladaptive decision-making, such that choices are biased toward excessively risky options. OBJECTIVE: The purpose of this study was to investigate the effects of a range of drugs of abuse on risky decision-making. METHODS: Male Long-Evans rats were trained in the Risky Decision-Making Task, in which they chose between two levers, one which produced a small, "safe" food reward and the other which produced a large, "risky" food reward. The large reward was accompanied by the risk of a mild footshock, the probability of which increased over the course of each test session (0%, 25%, 50%, 75%, and 100%). RESULTS: Nicotine (0.6 mg/kg) and amphetamine (1.5 mg/kg) caused a significant decrease in choice of the large risky reward (decreased risk taking). Diazepam (1.0 mg/kg) caused a significant increase in choice of the large risky reward (increased risk taking), whereas morphine (3.0 mg/kg) caused only a trend toward increased choice of the large risky reward. Ethanol had no effect on choice behavior. CONCLUSIONS: These results show that acute administration of drugs of abuse can modulate risk taking in a drug-specific manner, either increasing or decreasing preference for highly rewarding, but risky, options. PMID- 21638223 TI - Extreme sensitivity to acenocoumarol therapy in patient with both VKORC.-1639 A/A and CYP2C9*1/*3 genotypes. AB - The initiation phase of vitamin K antagonist (VKA) is a challenging and demanding process that can result in adverse event such as bleeding. Dosing is influenced by a variety of acquired factors, while another factor that is associated with the optimal dose is the presence of certain genetic variants. We describe a 73 years old male who required extremely low dose of acenocoumarol (0.33 mg/day) to reach the target INR of 2-2.5. During initiation of VKA he started with usually recommended doses of acenocoumarol: 4 mg/day for the first 2 days and 3 mg/day during next 2 days. On fifth day of initiation, massive haematuria occurred and INR level of 10.5 was recorded. Patient was transfused with three doses of fresh frozen plasma in order to stop the bleeding. Acenocoumarol dose was gradually reduced after every INR laboratory monitoring and finally a dose of 0.33 mg/day was required for maintain stable anticoagulation. The genotyping results performed after introducing of VKA showed the patient was homozygous for vitamin K epoxide reductase (VKORC1) c.-1639 G>A, and heterozygous for cytochrome P450 2C9 (CYP2C9)*3 which pointed to extreme sensitivity to acenocoumarol. Our case supports the need for prospective genotyping of CYP2C9 and VKORC1 prior to initiation of VKA where pharmacogenetics, as a good predictor of extreme sensitivity to VKA, could help to tailor optimal VKA dose. PMID- 21638224 TI - Elevated plasma gas6 levels are associated with venous thromboembolic disease. AB - Growth arrest-specific 6 (gas6), a novel vitamin K-dependent protein, has been demonstrated to have a role in thrombus stabilization as gas6 null mice are resistant to lethal venous and arterial thrombosis. However, the association between gas6 and venous thromboembolism has not been elucidated in humans. The present study aims to assess the role of gas6 in human venous thromboembolic (VTE) disease. Using a highly specific ELISA method, we measured plasma levels of gas6 in plasma samples obtained from 279 patients with VTE and 79 healthy volunteers. Medication history, comorbid conditions and VTE characteristics were documented. Mean gas6 levels were higher in patients with VTE as compared to healthy volunteers, being 46 +/-11 ng/ml and 35 +/-6.4 ng/ml respectively (P < 0.001). Odds ratios (OR) for VTE given elevated (>=90th percentile of healthy volunteers) gas6 levels were estimated in regression models in the whole study population. After adjustment for age, sex, medications and comorbidity, subjects with elevated gas6 had an increased risk of VTE (OR of 16.3 (95% CI 5.8-45.7, P < 0.001) compared to those with lower levels of gas6. This association remains significant even among patients with a comparable age distribution. Among patients with VTE, mean gas levels showed a trend of higher levels in those with more extensive thrombi. There was no correlation between elevated gas6 levels and recurrent VTE. In conclusion, we demonstrate an association between VTE and elevated gas6 levels consistent with in vivo murine models of thrombosis. This constitutes a potential novel mechanism for thrombosis in humans and may aid in the understanding of the pathophysiology of VTE. PMID- 21638225 TI - Persistently elevated prostate-specific antigen at six weeks after radical prostatectomy helps in early identification of patients who are likely to recur. AB - OBJECTIVE: To determine the prognostic factors of biochemical recurrence in patients who failed to achieve an undetectable prostate-specific antigen (PSA) after radical prostatectomy (RP) for prostate cancer. MATERIALS AND METHODS: We reviewed data on 240 men who underwent RP as first-line treatment and who had a PSA assay available at 6 weeks after surgery. Persistent detectable PSA was defined as a PSA level >= 0.1 ng/ml at 6 weeks after surgery. RESULTS: Overall, 83 men presented persistently elevated PSA after RP and 81 had a biochemical recurrence. Median follow-up was 44 months. In univariate analysis, these factors were associated with biochemical recurrence: preoperative PSA level (P < 0.0001), biopsy and pathologic Gleason score (P < 0.001), capsular involvement (P = 0.0001), positive surgical margins (P < 0.0001), pathological stage >= T3 (P = 0.0001), and detectable post-operative PSA >= 0.1 ng/ml (P = 0.0001). In a multivariate analysis, only the detectable post-operative PSA level >= 0.1 ng/mL (P = 0.001), positive surgical margins (P = 0.002), and pathological stage >= T3 (P < 0.001) were significant. The individual, five-year, PSA-free survival rate for men with post-operative PSA <0.1 ng/ml and >= 0.1 ng/ml were 59 and 42%, respectively (P < 0.001). CONCLUSIONS: A majority of patients who failed to achieve an undetectable PSA after surgery had a subsequent biochemical recurrence in the outcome. A systematic PSA assay 6 weeks after RP could be useful to early identify patients who are likely to recur. PMID- 21638226 TI - The black gender gap in educational attainment: historical trends and racial comparisons. AB - It is often asserted that the gender gap in educational attainment is larger for blacks than whites, but historical trends comparing the black and white gender gap have received surprisingly little attention. Analysis of historical data from the U.S. census IPUMS samples shows that the gender gap in college completion has evolved differently for whites and blacks. Historically, the female advantage in educational attainment among blacks is linked to more favorable labor market opportunities and stronger incentives for employment for educated black women. Blacks, particularly black males, still lag far behind whites in their rates of college completion, but the striking educational gains of white women have caused the racial patterns of gender differences in college completion rates to grow more similar over time. While some have linked the disadvantaged position of black males to their high risk of incarceration, our estimates suggest that incarceration has a relatively small impact on the black gender gap and the racial gap in college completion rates for males in the United States. PMID- 21638228 TI - Izmir mental health survey for gene-environment interaction in psychoses (TurkSch): objectives and methodology. AB - OBJECTIVE: To describe the objectives and design of a multistage study on the prevalence and risk factors of mental health problems, in particular psychotic disorders and psychotic symptoms, by assessing a probabilistic sample of household residents in the Izmir Metropolitan Area, aged 15-64 years. METHOD: The study included three different observation frames. In the first stage, of which methodology is in the focus of this paper, the primary screening instrument was the Composite International Diagnostic Interview 2.1, which assesses lifetime and/or last one year occurrence of several DSM-IV disorders. Second, a parallel survey of social capital of administrative wards was conducted in the same neighbourhoods. Third, a nested-case control study was performed to study effects of genetic and environmental risk factors on wide psychosis phenotype. RESULTS: A total of 4011 males and females were contacted through a multistage clustered area probability sample of administrative neighbourhoods and households, covering 9 districts and 302 neighbourhoods. The response rate for the first stage was 76.5% in 5242 eligible households. Respondents were interviewed at home in 2008 for the screening of included mental health problems. The screened disorders were mood disorders (last 1 year), schizophrenia and other non-affective psychotic disorders (lifetime), and dependence and abuse of psychoactive substances (lifetime). CONCLUSION: Although cross-sectional in nature, the TurkSch has a unique study design and yields data of high quality. This enables study of the prevalence, risk and higher-order interactions underlying ill-health in the Turkish population, with a specific focus on psychosis. PMID- 21638229 TI - Depression and anxiety levels and self-concept characteristics of adolescents with congenital complete visual impairment. AB - OBJECTIVE: Previous studies have reported that visual impairment can affect the mental health of children and adolescents. The aim of this study is to investigate the depression and anxiety levels and the self-concept characteristics of adolescents with congenital complete visual impairment. METHOD: This is a cross-sectional study. 40 adolescents with congenital complete visual impairment studying in a specialized primary school for visual impairment, and 40 sighted adolescents were included in the study. Both groups were matched in terms of age, gender and socio-economic status. The mean age of the adolescents in both groups was 12.82+/-1.17. The Children's Depression Inventory, Piers-Harris Children's Self-Concept Scale, Spielberger Trait Anxiety Inventory for Children and sociodemographic form were used in the study. The mean scores of the scales obtained from both groups were compared using the Mann-Whitney U test. RESULTS: The difference between the two groups was not statistically significant either in terms of depression scores or in terms of total scores; the happiness, physical appearance, popularity, behavior and adjustment subscales scores of the Piers-Harris Children's Self-Concept Scale. The intellectual and school-status subscale scores of the adolescents with visual impairment were significantly higher than those of the controls. Anxiety levels of the adolescents with visual impairment were significantly higher when compared with sighted adolescents. CONCLUSION: These results indicate that the depression levels and self-concept characteristics of adolescents with visual impairment are similar to those of sighted adolescents, whereas the anxiety levels of the adolescents with visual impairment are significantly higher than those of the sighted ones. PMID- 21638230 TI - The relationship between traumatic childhood experiences and proclivities towards substance abuse, self-esteem and coping strategies. AB - OBJECTIVE: The aim of this study was to compare people with substance use disorder with healthy controls in terms of childhood abuse, proclivity towards substance abuse, coping skills and self-esteem as well as the correlation between experiences of abuse in childhood and these variables. METHOD: The study group included 41 subjects diagnosed with substance use disorder, who had been sentenced under the respective laws as a result of crimes relating to substance use and possession, and the control group. A sociodemographic Data Form, SCID-I, Childhood Trauma Questionnaire, COPE, Substance Abuse Proclivity Scale and Rosenberg Self-Esteem Scale were applied to all participants. RESULTS: Childhood trauma history was observed to be more common in the study group than in the control group. When the childhood trauma questionnaire was evaluated, the scores for physical abuse were found to be significantly higher in the study group. COPE subscale scores for mental disengagement, focusing on problems and expressing emotions, active coping, coping through religion and emotional social support usage were significantly lower in the study group. The study group's results on the Substance Abuse Proclivity Scale were found to be higher than those of the control group. On the Rosenberg Self Respect Scale, the study group's scores were higher while the control group was more likely to have high self respect. CONCLUSION: People with substance use disorder are more likely to have a childhood history of physical abuse, higher proclivity towards substance abuse and lower self esteem. The level of abuse increases the level of emotion-based coping while decreasing levels of problem-based coping. There is support for the view that that traumatic childhood experiences are one of the psychosocial risk factors related to, although not specific to substance use. PMID- 21638231 TI - Adolescents living in orphanages in Ankara: psychological symptoms, level of physical activity, and associated factors. AB - AIM: Adolescents living in orphanages are at a disadvantage with respect to mental health. The aim of this study was to assess the distribution of psychological symptoms and their association with the level of physical activity (PA) in adolescents living in orphanages. METHOD: The study group consisted of 13 16-year-old adolescents (N=166) living in orphanages in Ankara, Turkey. Data were collected cross-sectionally in 2008 via questionnaires, including the Brief Symptom Inventory and Kiddo-KINDL Health-Related Quality of Life Questionnaire. Descriptive statistics were used to summarize data, whereas chi-square, ANOVA, Mann-Whitney U, Kruskal-Wallis, and t tests were used to compare groups. RESULTS: Median age of the participants was 16.0 years and 65.7% were male. Female gender, not going to school or work, dissatisfaction with school, contact with the family, chronic disease, chronic medication use, sleep problems, regular tobacco use, chronic disease in the family, and low quality of life score were associated with increased risk (GSI-Global Symptom Index>1 SD) for mental disorders. Physically active adolescents' use of tobacco, alcohol, other substances, and medications, as well as GSI and depression scores were lower and their quality of life scores were higher than those of their less active counterparts. CONCLUSION: Prevalence of psychological symptoms in adolescents living in orphanages were higher than in the general adolescent population. Physically active adolescents' mental health indices and abstinence behaviors with regard to tobacco, alcohol, and substances were more favorable. Encouraging adolescents to participate in sports and improving sports facilities in orphanages are interventions that can promote mental health. PMID- 21638232 TI - The reliability and validity of the Resilience Scale for Adults-Turkish Version. AB - OBJECTIVE: Resilience is as a dynamic process related to an individual's capacity to cope with difficult or stressful experiences and the ability to psychologically overcome adversity. The aim of this study was to examine the reliability and validity of the Resilience Scale for Adults-Turkish Version. METHOD: The Resilience Scale for Adults was first translated into Turkish, and was then back translated. Subsequently, the questionnaire was administered to 350 students and 262 employees. The internal consistency and test-retest reliability of the scale were determined, and criterion-dependent validity and confirmative factor analysis were conducted using Amos v.16.0. RESULTS: Factor analysis of the scale confirmed the fit of the original's 6-dimensions: perception of self, perception of future, structured style, social competence, family cohesion, and social resources (c2=1104, df=480, c2/df=2.3; RMSEA=0.055; TLI=0.90; CFI=0.91). The 6-dimensions structure explained 53.5% of the total variance. The Social Comparison Scale and Locus of Control Scale were used to determine the criterion dependent validity of the scale. Alpha coefficients for the sub-dimensions of the scale ranged from 0.66 to 0.81 and the test-retest reliability of the factors ranged from 0.68 to 0.81. CONCLUSION: The present findings show that the Resilience Scale for Adults-Turkish Version exhibited acceptable levels of reliability and validity in the study samples. PMID- 21638233 TI - Fear or disgust? The role of emotions in spider phobia and blood-injection-injury phobia. AB - According to the literature, it is assumed that fear and anxiety are basic emotions in anxiety disorders. Many recent studies report that disgust, as well as fear, has an important role in the etiology and maintenance of anxiety disorders. Evaluation of the role of disgust in anxiety disorders has led the theoretical and empirical literature in a new direction, beyond the traditional emphasis on fear. Most of this basic research has focused on specific phobias, such as blood-injection-injury phobia and spider phobia. Findings obtained from evaluation of physiological and cognitive processes, and subjective and behavioral experiences clearly show that in addition to fear, emotional reactions to phobic stimuli also include disgust; however, empirical studies show that disgust and fear have different relative impacts on specific phobias. To illustrate, individuals experience disgust as the basic emotion in blood injection-injury phobia, whereas both fear and disgust are experienced in spider phobia. Nevertheless, it is concluded that fear has a more fundamental role in the latter. Yet, research indicates that basic emotions different from those identified from neural structures or physiological responses, such as heart rate, can be identified if facial expressions and cognitive appraisals are taken into account. In the present review the role of fear and disgust in blood-injection injury phobia vs. spider phobia are discussed, based on the relationship between the phobias and disgust sensitivity, disgust as part of phobic responses, and disgust-motivated avoidance behavior. PMID- 21638234 TI - Risperidone treatment for polydipsia and hyponatremia in schizophrenia: a case report. AB - Psychogenic polydipsia with associated hyponatremia is a potentially fatal condition observed in patients with chronic psychiatric illness, especially schizophrenia. Recognition and management of this condition are difficult, as patients are uncooperative and secretive about their water intake, but are important in terms of the associated complications. Different strategies, including involuntary fluid restriction and use of various pharmacological agents, such as demeclocycline, propranolol, captopril, and naloxone, have been used for the treatment of this condition with inconsistent results. Antipsychotics have also been used in the treatment of polydipsia; however, their role is not clear as there are reports of antipsychotics both improving and causing polydipsia. Typical antipsychotics have been associated with exacerbation of polydipsia, whereas clozapine has been associated with its improvement. The efficacy of risperidone in the treatment of this condition is controversial, as negative results have been reported. Herein we present a schizophrenia case with polydipsia and hyponatremia that was successfully treated with risperidone. PMID- 21638235 TI - Physical deformity associated with psychotic symptoms in two patients with schizophrenia. AB - Movement disorders and related physical deformities may sometimes be observed in patients with chronic psychotic disorders. In this article, we present the case of two patients with physical deformities associated with chronic psychotic disorders. In the first case, the patient had never sought psychiatric care despite her long-standing psychiatric disorder. The patient, diagnosed with disorganized schizophrenia, developed cervical kyphosis, due to her constant neck flexion posture. The other patient had been undergoing treatment for a long period under the diagnosis of paranoid schizophrenia. In the latter case, peroneal nerve injury and dropped foot had developed due to a constant crossing of the legs. Physical deformity may also develop as a result of physical inactivity-hypokinesia, a fixed body posture, and postural disorders in chronic psychotic patients. Due attention should be given to physical symptoms in this group of patients and physical deformities should be treated alongside the patient's psychotic symptoms. PMID- 21638236 TI - [Professor Dr. Dieter Haussinger on his 60th birthday]. PMID- 21638237 TI - Cell volume, the serum and glucocorticoid inducible kinase 1 and the liver. AB - In virtually all cells including hepatocytes cell volume regulation is accomplished during cell swelling by cellular ion release (activation of K (+) channels and/or anion channels, KCl-cotransport, parallel activation of K (+)/H (+) exchange and Cl (-)/HCO (3)(-) exchange) and following cell shrinkage by cellular ion uptake (activation of Na (+), K (+), 2Cl (-) cotransport, Na (+)/H (+) exchange in parallel to Cl (-)/HCO (3)(-) exchange and Na (+)-channels). Moreover, cell shrinkage triggers the cellular accumulation of organic osmolytes (e. g., myoinositol, betaine, phosphorylcholine, taurine). Cell volume is a powerful regulator of hepatic metabolism. Cell shrinkage stimulates and cell swelling inhibits proteolysis and glycogenolysis. Moreover, cell volume influences the generation of and sensitivity to oxidants. Cell volume regulatory mechanisms furthermore do play a role in fibrosing disease. Kinases stimulating cell volume regulatory mechanisms include the serum and glucocorticoid inducible kinase SGK1, which is expressed in the liver, is genomically up-regulated by cell shrinkage, stimulates a wide variety of channels and transporters including Na (+), K (+), 2Cl (-) cotransport and Na (+)/H (+) exchange and is known to participate in the stimulation of fibrosis. Accordingly, excessive SGK1 expression is observed in liver cirrhosis. The case is made that SGK1 participates in the regulation of liver cell volume and thus in the regulation of hepatic metabolism. PMID- 21638238 TI - [Increasing number and improved survival of patients with hepatocellular carcinoma from 1988 to 2007: data of a German university clinic]. AB - BACKGROUND AND OBJECTIVES: Hepatocellular carcinoma (HCC) ranks sixth regarding prevalence and third regarding mortality among malignant tumours worldwide. The aim of the present study was to determine changes of clinical-epidemiological parameters and survival rates during two decades. PATIENTS AND METHODS: A total of 441 consecutive patients with HCC admitted to the University Clinic Dusseldorf between January 1988 and December 2007 were included. For comparison, this time period was divided into two decades (1988 - 1997 and 1998 - 2007). RESULTS: The number of newly diagnosed HCCs has tripled in the years 1998 - 2007 compared to the years 1988 - 1997. HCV-associated HCCs increased from 28 % in the years 1988 1997 to 38 % (p < 0.05) in the years 1998 - 2007. Tumour size, Okuda and BCLC stages decreased during the observation period (both p < 0.001 and p < 0.05). Median overall survival improved during the observation period from 6 [95 % CI: 4.83 - 7.17] to 9 months ]95 % CI: 7.31 - 10.69]; p < 0.0001) as did the 1-year and 5-year survival rates from 22 % to 42 % (p < 0.019) and from 0 % to 9 % (p < 0.001), respectively. The proportion of treated patients compared to patients with best supportive care as well as the proportion of patients receiving a multimodal therapy compared to patients with a single treatment regimen increased in the second decade (55 % vs. 79 %: p < 0.005; 5.4 % vs. 23 %: p < 0.0001). Multimodal therapy was an independent predictor for prolonged survival in a multivariate analysis including Child-Pugh score, BCLC stage, tumour size, and gender (odds ratio 2,77; 95 % CI: 1.44 - 5.31). CONCLUSION: Improved screening as well as broader and improved treatment options may have contributed to the increasing survival rates. PMID- 21638239 TI - Cholestatic liver diseases from child to adult: the diversity of MDR3 disease. AB - The phospholipidfloppase MDR3 (gene symbol: ABCB4) is expressed in the canalicular membrane of hepatocytes and mediates the biliary excretion of phosphatidylcholine, which is required for the formation of mixed micelles in bile. Several mutations of ABCB4 have been identified, which cause cholestatic liver diseases of varying severity including progressive familial intrahepatic cholestasis type 3 (PFIC-3), intrahepatic cholestasis of pregnancy (ICP) and the low phospholipid associated cholelithiasis syndrome (LPAC). Here, we report on four new (S1076N; L 23Hfs16X; c.286 + 1G > A; Q 1181E) and one known (S27G) MDR3 mutations in eight patients of three families. The patients presented with a wide spectrum of liver diseases. The clinical presentation and decisive laboratory findings or the association to a trend-setting family history led to the identification of the genetic background in these patients. Even the same mutation may be associated with varying disease progression. PMID- 21638240 TI - Use of isolated liver perfusion in metabolic studies: ground-laying work in experimental hepatology. AB - Modern metabolic research in hepatology employs non-invasive techniques of the whole organism, and it includes studying the intact organ. Following recent decades of efforts in culturing isolated cells and studying their properties separately, it has become clear that the spatiotemporal organisation of different cell types in a tissue requires studies using models of the intact organ or tissue. This applies particularly to the liver as the major organ of metabolic transformation and activity. The present brief article attempts to illustrate the advantages and limitations of such approaches, focusing on selected aspects of ammonia/urea and glutamine metabolism as an example. PMID- 21638241 TI - [Endoscopic therapy for leaks in the gastrointestinal tract, the bile ducts and the pancreas]. AB - Surgery has been the mainstay of therapy in patients with gastrointestinal perforations, leakage or fistulas. New techniques for endoscopic closure of gastrointestinal perforations provide tools for an effective treatment by less invasive procedures. Temporary placement of covered self-expanding stents is an established therapy for oesophageal perforations and anastomotic leaks. Using conventional endoclips small perforations and leaks in the oesophagus and gastrointestinal tract may be closed. With the new over-the-scope-clips a more effective endoscopic full wall closure is possible in the upper gastrointestinal tract and the rectum. Endoscopically guided endoluminal vacuum therapy using polyurethane sponges is an established method for treating rectal leaks and is now increasingly used also in oesophageal leaks. Biliary leakage following endoscopic or surgical interventions is effectively treated with temporary bile stenting in most cases, but closure using metal stents or coiling may be necessary. Pancreatic leaks are a major therapeutic problem and may require multimodal therapies. PMID- 21638242 TI - [German S 3-guideline "ascites, spontaneous bacterial peritonitis, hepatorenal syndrome"]. PMID- 21638243 TI - [Method report: German S3-guideline "ascites, spontaneous bacterial peritonitis, hepatorenal syndrome"]. PMID- 21638244 TI - Pharmacokinetics, bioavailability, and tissue distribution of magnolol following single and repeated dosing of magnolol to rats. AB - Magnolol (M) is a polyphenol antioxidant abundant in the bark of Magnolia officinalis Rehder & E. Wilson, a popular Chinese herb. To understand the pharmacokinetics and bioavailability of M, Sprague-Dawley rats were intravenously injected with a bolus of M (20 mg/kg) and orally given a single dose and seven doses of M (50 mg/kg). Blood samples were withdrawn via cardiopuncture at specific times. Organs including the liver, kidney, brain, lung, and heart were collected at 30 min after the 7th oral dose. The serum and tissue specimens were assayed by HPLC before and after hydrolysis with beta-glucuronidase and sulfatase. The results showed that after intravenous bolus, the systemic exposure of magnolol glucuronides (MG) was comparable with that of M while after oral administration, magnolol sulfates/glucuronides (M S/G) were predominant in the bloodstream. Conversely, M was predominant in the liver, kidney, brain, lung, and heart. Among the studied organs, the liver contained the highest concentrations of M and MG. In conclusion, M S/G was the major form in circulation, whereas M was predominant in the liver, kidney, brain, lung, and heart after oral administration of M; among these organs, the liver contained the highest concentrations of M and MG. PMID- 21638245 TI - A new cytotoxic beta-carboline alkaloid from Galianthe thalictroides. AB - A cytotoxicity-guided fractionation of the roots of Galianthe thalictroides afforded a new beta-carboline alkaloid, 1-(hydroxymethyl)-3-(2-hydroxypropan-2 yl)-2-(5-methoxy-9H- beta-carbolin-1-yl)cyclopentanol, which exhibited strong cytotoxic activity against three human cancer cell lines (MCF-7, 786-0, and UACC62). Its structure was established on the basis of 1D- and 2D-NMR spectroscopic techniques supported by HRMS data. PMID- 21638246 TI - Therapeutic arteriogenesis in peripheral arterial disease: combining intervention and passive training. AB - The prevalence of peripheral arterial disease (PAD) is on the rise in an aging population, significantly affecting quality of life, morbidity and mortality. Besides medical treatment and surgical or interventional revascularization, supervised exercise programs are a primary treatment modality for PAD. Training may significantly increase pain-free walking time (+ 180 %) while avoiding the associated complications of (repeated) invasive revascularization. Training effects rely on an improvement of risk factor management, muscle function, economy of movement, hemorheology, vascular growth and collateral vessel growth. Exercise training upregulates pulsatile fluid shear stress on the vascular endothelium, prompting an improvement of endothelial function (eNOS, NO) and an outgrowth of preexistent collaterals (arteriogenesis) to functional conductance arteries outside the ischemic area at risk. However, the necessary intense minimum training intervals compromise patient compliance, and the impaired functional status of many PAD patients limits active exercise training. Strategies are necessary to a) increase training compliance, b) make the benefits of exercise training available to patients unable to exercise actively and c) therapeutically enhance the adaptive growth of biological bypasses (arteriogenesis). A modified form of ?passive exercise training? derived from enhanced external counterpulsation (low-pressure ECP) which was originally developed for the therapy of heart failure, may prove to be an option for this group of patients. Therefore, this review article suggests a tailored combination therapy, consisting of a facilitating revascularization procedure to restore arterial inflow, succeeded by supervised exercise training, which has yielded promising therapeutic results in clinical trials. Further studies, using appropriate imaging methods and controls, are under way to (a) establish the efficacy of low-pressure EECP in PAD patients and (b) to directly correlate training-induced improvements of collateral flow to the functional improvements seen with exercise training. PMID- 21638247 TI - Multidisciplinary management of vascular access for haemodialysis: from the preparation of the initial access to the treatment of stenosis and thrombosis. AB - In February 2008 a multidisciplinary study group was established in Germany to improve the treatment of patients with potential vascular access problems. As one of the first results of their work interdisciplinary recommendations for the management of vascular access were provided, from the creation of the initial access to the treatment of complications. As a rule the wrist arteriovenous fistula (AVF) is the access of choice due to its lower complication rate when compared to other types of access. The AVF should be created 3 months prior to the expected start of haemodialysis to allow for sufficient maturation. Second and third choice accesses are arteriovenous grafts (AVG) and central venous catheters (CVC). Ultrasound is a reliable tool for vessel selection before access creation, and also for the diagnosis of complications in AVF and grafts. Access stenosis and thrombosis can be treated surgically and interventionally. The comparison of both methods reveals advantages and disadvantages for each. The therapeutic decision should be based on the individual patients' constitution, and also on the availability and experience of the involved specialists. PMID- 21638248 TI - Expression of P-selectin, von Willebrand and endothelin-1 after carotid artery stenting. AB - BACKGROUND: This study was designed to investigate the impact of carotid artery stenting (CAS) on plasma levels of P-selectin, von Willebrand (vWF) and endothelin-1. PATIENTS AND METHODS: Sixty-seven patients who received CAS were divided into group 1 (one stent for a simple lesion, n = 38) and group 2 (two stents for complex lesions, n = 29). The levels of P-selectin, vWF and endothelin 1 were measured before CAS, 1 h, 6h, 24 h and 2 weeks after the stenting. RESULTS: Sixty-one patients completed one-year follow up. Restenosis was noted in 14 (23 %) patients, among these three (4.8 %) had a restenosis of > 50 % of the vascular lumen. In all patients, the levels of P-selectin, vWF and endothelin-1 increased immediately after CAS (P < 0.05 or < 0.01). The levels of vWF and endothelin-1 in group 2 were higher than in group 1 (P < 0.05 or 0.01). There was no significant difference in P-selectin and endothelin-1 between the restenosis and non-restenosis group (P > 0.05). The 24 h vWF in patients with restenosis were higher than in non-restenosis group (P < 0.05). CONCLUSIONS: CAS results in a significant increase in plasma P-selectin, vWF and endothelin-1. The post-CAS levels of P-selectin, vWF and endothelin-1 are related to the extent of endothelial injury. Whether they are associated with restenosis 12 months after the treatment requires further investigation. PMID- 21638249 TI - The venous return simulator: comparison of simulated with measured ambulatory venous pressure in normal subjects and in venous valve incompetence. AB - BACKGROUND: To compare results of numerical simulation of lower limb venous return with those of in vivo measurements, in normal subjects, and those with venous incompetence. PATIENTS AND METHODS: the venous return simulator (VRS) is a mathematical model which takes into account architecture, dimensions, and compliance of the venous network, blood viscosity, valve function, and external pressures (muscular contraction, compression stockings). Using the laws of hydrodynamics, it provides calibres, pressures and flows throughout the network. Ambulatory venous pressure (AVP) computed for some theoretical examples of superficial and /or deep venous incompetence has been compared to in vivo values reported in literature. RESULTS: In a normal subject, computed AVP was 33 mmHg during walking and 30 mmHg with tiptoe exercise; the range of conventionally measured AVP is 20.6 - 27.9 mmHg during walking, and 29 - 32.5 mmHg during tiptoe exercise; In the case of great saphenous vein (GSV) incompetence, computed AVP was 34 or 57 mmHg, according to whether the distal GSV was competent or not. The range of AVP measured in superficial venous insufficiency is 27.6 - 61 mmHg, all but one of the published values lie between the low computed value corresponding to a short reflux and the high computed value due to a long distance reflux. AVP computed in two cases of deep venous incompetence was 44 and 71 mmHg, according to the extent of devalvulation, as compared with the 60 mmHg reported in one clinical study In patients with extensive combined incompetence, computed AVP was 75 mmHg, whilst the range of conventionally measured values was between 62 and 84 mmHg. CONCLUSIONS: the good agreement between computed and measured AVP in different cases of valve incompetence indicates that the VRS is quite a realistic model, with the potential to simulate the results of surgery or compression therapy. PMID- 21638250 TI - Patterns of extracranial involvement in newly diagnosed giant cell arteritis assessed by physical examination, colour coded duplex sonography and FDG-PET. AB - BACKGROUND: The clinical spectrum of giant cell arteritis (GCA) varies from classical temporal arteritis (TA) to generalized large vessel GCA (LV-GCA) and fever of unknown origin (FUO). Extent and distribution of extracranial involvement in these different presentations of GCA is not well known, and its detection may depend on the choice of vascular imaging. PATIENTS AND METHODS: In 24 patients with newly diagnosed GCA we systematically evaluated the presence and distribution of extracranial involvement by physical examination, duplex sonography (DS), and FDG-PET. Analysis of FDG-PET results was performed in comparison with 18 age-matched control-subjects scanned for oncological indications. RESULTS: Initial clinical diagnosis was TA in 11 patients, LV-GCA in 8 patients, and FUO in 5 patients. Clinically detectable arterial obstruction was present in 2 patients (18 %) with TA (only upper extremity), all patients with LV GCA (upper and lower extremities) and no patient with FUO. Upper and/or lower limb large vessel vasculitis was detectable by DS in 45 % of the patients with TA and in 100 % of the patients with LV-GCA or FUO. FDG-PET confirmed upper extremity involvement in all affected patients, but had a very low specificity for lower limb involvement due to concomitant arteriosclerosis in these elderly patients. Aortitis was detectable by FDG-PET in 27 % of patients with TA and 75 - 80 % of patients with LV-GCA or FUO. CONCLUSIONS: The combination of thorough clinical examination and DS is able to detect symptomatic as well as asymptomatic large vessel involvement in a large proportion of patients with newly diagnosed GCA. Distribution and manifestation of large vessel involvement differs between classical TA and LVGCA or FUO. FDG-PET provided only limited additional information and did not change the clinical diagnosis in any patient. PMID- 21638251 TI - Increased health care utilization by patients with peripheral arterial disease in primary care: data from the CONTENT study. AB - BACKGROUND: Due to demography the proportion of elderly patients with peripheral arterial disease (PAD) in primary care settings is continuously growing. The study aimed to compare general practitioner care of patients with PAD with those without PAD. PATIENTS AND METHODS: The analyzed data are derived from the general practice morbidity research network CONTENT (CONTinuous morbidity registration Epidemiologic NETwork). A sample of 480 patients with ensured PAD diagnosis was extracted from a total of 89516 steadily registered primary care patients. Matched pair analysis with the propensity score was used statistically to compare the data of each PAD patient with data of three associated control patients. RESULTS: During a two-year period, PAD patients had significantly more consultations (20.1 vs. 18.0; p < 0.0001), more referrals to specialists (5.5 vs. 3.5; p < 0.0001) and a higher rate of hospitalization (0.3 vs. 0.1; p < 0.0001) than corresponding controls. The analysis of reasons for doctor-patient encounters showed a significantly higher number of disordered sleep, alcohol abuse, dyspnea and tobacco abuse among PAD patients compared to patients without PAD. The ranking of the most frequent reasons for referral to specialists suggested significantly more diagnoses related to cardiovascular disease in PAD patients. In the PAD cases, 20.4 % of all inpatient treatments were related to complications of the vascular disease, whereas no patient of the control group had to be hospitalized due to vascular disease problems. CONCLUSIONS: The CONTENT episode-based registration presents a realistic and detailed picture of health care utilization of PAD patients. They overuse medical services in primary care compared to corresponding unaffected patients. This is associated with deficits in their life-style behavior, increased cardiovascular morbidity and increased inpatient care. PMID- 21638252 TI - Bipolar radiofrequency induced thermotherapy and 1064 nm Nd:Yag laser in endovenous occlusion of insufficient veins: short term follow up results. AB - BACKGROUND: The aim of this study is to report our results in main stem vein closure using the bipolar radiofrequency induced thermotherapy (RFITT) system and the 1064 nm Nd:Yag laser. PATIENTS AND METHODS: 44 incompetent main stem veins (37 great saphenous veins, one lesser saphenous vein, and 6 anterior accessory saphenous veins) in 29 patients were treated using RFITT. 53 incompetent main stem veins (45 great saphenous veins, 4 lesser saphenous veins, and 4 anterior accessory saphenous veins) in 43 patients were treated endovenously with 1064 nm Nd:Yag laser. All patients underwent postoperative duplex scanning within a month after procedure, as well as a short interview regarding postoperative discomfort. RESULTS: In main stem veins treated with RFITT, the success rate within the first month was 86,4 % (38 out of 44 veins). Complete failure rate was 13,6 % (6 out of 44 veins). In 53 main stem veins treated by 1064 nm Nd:Yag laser, the success rate was 100 %, consisting of 98,1 % complete success (52/53 veins), and 1,9 % partial success (1/53 veins). None of the patients treated with RFITT experienced postoperative adverse effects, whereas 13/43 (30,2 %) patients treated with laser had to use oral analgesics after the treatment, and 21/43 (48,8 %) patients reported transient skin changes, such as bruising or skin redness. CONCLUSIONS: RFITT system was fairly efficient in the short term for closure of main trunk veins, whereas longer term results are still scarce. Postoperative side effects of RFITT were minimal. 1,064 nm Nd:Yag laser, according to short term results, proved to be very effective for main stem vein closure. Postoperative side effects related to 1064 nm Nd:Yag endovenous laser treatment proved to be minor, transient, and acceptable. PMID- 21638253 TI - Hybrid procedures for the treatment of multi-focal ipsilateral internal carotid and proximal common carotid or innominate artery lesions. AB - We report three cases with concomitant ipsilateral proximal common carotid and internal carotid artery stenosis treated in one stage with carotid endarterectomy and retrograde primary stenting of the common carotid artery. The internal carotid artery was clamped during stenting to avoid cerebral embolization. All procedures were successfully completed and all patients remain asymptomatic at 18 months follow up. The one-stage hybrid approach appears to be a safe and effective procedure for the treatment of ipsilateral multifocal significant lesions. PMID- 21638254 TI - Endovascular repair of a spontaneous ilio-iliac fistula presenting as pulmonary embolism. AB - Spontaneous rupture of a common iliac artery aneurysm into the common iliac vein is a rare phenomenon. We report the case of a 68 year old man admitted with acute cardiac failure and massive pulmonary embolism as a complication of a spontaneous ilio-iliac fistula, secondary to aneurysmal rupture. The aneurysm was successfully excluded using an aorto-uni-iliac stent graft. No complications were noted at 9 months follow-up. Arteriovenous fistulae should be considered in patients with aortic or iliac aneurysms who develop a pulmonary embolism or symptoms of venous congestion. Endovascular repair of these pathologies is a feasible therapeutic option; however long term results remain unknown. PMID- 21638255 TI - Popliteal artery thrombosis secondary to a tibial osteochondroma. AB - Popliteal artery thrombosis may present as a complication of an osteochondroma located in the vicinity of the knee joint. This is a case report of a 26-year-old man with symptoms of the right lower extremity ischaemia without a previous history of vascular disease or trauma. Plain radiography, magnetic resonance angiography and Doppler ultrasonography documented the presence of an osteochondrous structure of the proximal tibial metaphysis, which displaced and compressed the popliteal artery, causing its occlusion due to intraluminal thrombosis..The patient was operated and histopathological examination confirmed the diagnosis of osteochondroma. PMID- 21638256 TI - [Information needs of people suffering from venous leg ulcer. Expertise of people concerned as a basis of patient-centered information]. AB - Patient-centred information is orientated on the needs. The main objective of such information is coping with every-day life and learning to live with the chronic disease. Regarding the current knowledge, such information does not exist in German speaking countries for people suffering from venous leg ulceration. In order to generate the subject matter of such information, a literature search and a secondary analysis of 27 transcripts of interviews, given by people living with chronic leg ulceration, were conducted. Both analyses show the complex impact of leg ulceration on every-day live. The dimensions competences in every-day live, knowledge, techniques and skills and the meaning of family were generated. The secondary analysis points out that family members are an important part of the supporting system and are concerned by effects of chronic disease as well. After long-lasting disease-experience people consider themselves as experts. They obtain special competences in dealing with their disease and judge their experience-based knowledge higher than the knowledge of healthcare professionals. The participants describe individual concepts about their disease. Medical knowledge constitutes just one source of knowledge among others. People know about alternative medicine and use it. They feel that the healthcare professionals do not take them seriously in most cases. Patient-centred information can be a guide to people suffering from venous leg ulceration and their family members, but also a support for healthcare professionals to reflect their professional behaviour and understanding. PMID- 21638257 TI - [The development of an instrument for the "evaluation of hospices from the bereaved family members perspective"]. AB - Inpatient hospice settings as facilities of health services are obliged to the quality assurance and internal quality development in Germany. The meaning of the patient's satisfaction as one of the indirect indicators for the judgement of nursing and care quality is indisputable by now. However, a subjective evaluation of the hospices by guests is practically and ethically problematic. An alternative approach is to investigate the views of bereaved relatives and close friends after the guest's death. The present article describes the development of an inventory for the evaluation of inpatient hospice settings from the bereaved relatives' point of view in order to examine the satisfaction of family members with the end-of-life care their loved ones received. With help of the inventory the question should be answered how family members judge the end-of-life care which was given to them and their late family members by the hospice. The construction of the questions results from the basis of existing concepts to the assessment of the end-of-life care and an analysis of all identified instruments to the evaluation of inpatient hospice settings from the perspective of bereaved relatives. The development of the questionnaire enclosed the formation of an item pool, the item choice, the critical discussion of the questions in the body of experts as well as a standardised and cognitive pretest beside a comprehensive literature research. A five-dimension questionnaire was developed that integrates physical, psychological, social, spiritual, and organisational aspects of the care at the end of life. The instrument encompasses 53 items, predominantly closed questions. PMID- 21638258 TI - [Preventive measures for the prophylaxis of contractures in geriatric nursing. A systematic review]. AB - Contractures constitute a health problem in mobility-restricted geriatric patients. Contractures are clinically important due to their impact on functional outcome. Prophylactic interventions seem to be indispensable. We performed a systematic review studying the interventions used to prevent immobility-related contractures in the geriatric long-term care. The efficacy and safety of preventive methods was assessed. Electronic literature searches covered the databases PubMed, PEDro, CINAHL and the Cochrane Library (May 2010). Eligibility criteria for studies were: Investigation of an intervention aimed to prevent contractures, conducted in a geriatric and long-term care setting, inclusion of participants aged>65 years. The primary search focused on RCTs, systematic reviews und meta-analysis published between 1990 and May 2010 in English or German. The included studies were analysed and evaluated by one author while a second author checked the results. Methodological quality was critically evaluated using internationally accepted criteria. Eight studies met the inclusion criteria. Prophylactic interventions comprise mobility-encouraging and position-supportive interventions. Mobility-encouraging interventions aim to prevent contractures and immobility, for example offering range-of-motion exercises. Position-supportive interventions are transformations of motion and position, which are conducted with nurses' support. Due to limited methodological quality of these studies, the efficacy of certain measures remains unclear. Further studies on contracture prophylaxis investigating patient-relevant outcomes, interventions' adverse effects and costs are required. PMID- 21638259 TI - [Competence diagnostic in the field of nursing]. AB - The efficiency and effectiveness of educational programmes in the occupational field of nursing are only assessable through testing and comparing achieved learning results. In search of an appropriate diagnostic instrument, a comprehensive literature research in German-speaking countries concerning the current state of competence diagnostics in the occupational field of nursing had been conducted. Additionally, a similar research of international, professional journals had been done. The investigations lead to the result that a massive gap exists concerning national and international research of systematic, theoretically and empirically well-founded development of instruments measuring nursing competence. Finally, a conclusion for the development of instruments in nursing competence diagnostics is drawn. PMID- 21638261 TI - [Bluetongue disease: impact of the 2008 vaccination on fertility in supervised dairy herds]. AB - In June 2008 the compulsary nationwide vaccination against BTV-8 (Bluetongue virus serotype 8) was started. After a short time, several owners complained about undesirable effects of the vaccination on fertility and milk quality. Data from 47 dairy farms, regularly supervised by herd health practitioners, were analysed in order to clarify a possible connection between vaccination and fertility. Both vaccinations given each cow for basic immunization were evaluated according to their effects on conception rate and pregnancy. In model calculations the first vaccination had no significant effect on the first service conception rate (FCR), the all service conception rate (ACR) and on the abortion rate. The second vaccination led to a significantly reduced FCR when the cow was inseminated within 20 days of being vaccinated and to a significantly worse ACR when inseminated 10 days before or after vaccination. However, these individually established reductions of the insemination rate had only little influence on overall data. PMID- 21638262 TI - [The role of bacterial contamination of milking utensils and disinfecting solutions as a possible cause of clinical mastitis in dairy cows]. AB - Various instruments and utensils used during milking as well as teat dip solutions were examined for contamination with coagulase-negative staphylococci (CNS). The goal of this study was to investigate the relationship between contaminated fomites and udder infection in dairy cows. A total of 344 cows from ten dairy farms with the highest rate of clinical mastitis among the farms serviced by the Ambulatory Clinic of the University of Zurich were included in the study. Each farm was visited five times. All lactating cows, with the exception of those undergoing antibiotic treatment, were examined immediately before milking using the California Mastitis Test (CMT). A milk sample was collected from positive quarters. Items used to clean the udder, which included wood wool, paper towels and disinfecting towels as well as the milker's hands and the teat dip cup were swabbed for bacteriological examination. Water samples, samples of teat dip and cleaning solutions were also collected and cultured. Our results demonstrate that cleaning and disinfecting solutions have the potential to transmit udder pathogens and cause clinical mastitis. The most common CNS isolated from quarter samples were S. saprophyticus, S. sciuri and S. chromogenes, and the most common CNS isolated from utensils, cleaning and disinfecting solutions were S. fleuretii, S. vitulus, S. equorum, S. sciuri, S. haemolyticus, S. succinus and S. saprophyticus. PMID- 21638263 TI - [Hereditary eye dieseas in the Entlebucher Mountaindog in Switzerland: a retrospective study from 1999 to 2009]. AB - The aim of this study was to analyze the prevalence and the incidence of hereditary cataract and progressive retinal atrophy (PRA) in the Entlebucher Mountaindog (EMD) and to evaluate possible changes over time. In addition, the influence of selective breeding programs and DNA-testing for PRA was also investigated. Data of eye certifications for hereditary eye diseases was used. Between 1999 and 2009 a total of 798 ocular examinations were carried out in 285 EMD. 20.4 % had cataracts and 69 % of these were of the posterior polar type. PRA was diagnosed in 6.3 % of the dogs. Cataracts were diagnosed at 5.24 +/- 2.71 years (mean +/- standard error), while PRA was diagnosed at 4.93 +/- 1.32 years of age. The incidence of PRA and cataract showed a decreasing trend (p-value > 0.2) without being significant. PMID- 21638264 TI - [Treatment of atresia ani type I by balloon dilatation in 5 kittens and one puppy]. AB - Atresia ani is the most common anorectal anomaly in small animals. In the present study, an anal stricture (atresia ani type I) in five 3 to 8 weeks old kittens and one 4 month old puppy was treated by balloon dilation. In 4 kittens and the puppy the stricture was eliminated permanently and without complications by a single intervention. Only the smallest kitten with the most severe stenosis developed a rectal fistula as a complication of repeated balloon dilation, which necessitated surgical correction. Balloon dilation proved to be an efficient therapeutic method for anal atresia type I, and can be recommended as the treatment of choice. PMID- 21638265 TI - [Multiple tooth resorption in an Italian greyhound]. AB - An Italian greyhound was presented three times during a two-year period for dental prophylaxis due to periodontal disease. Clinical examination revealed lesions on several teeth. Radiographs revealed extensive resorptive root lesions. On histological examination, the presence of odontoclasts and signs of boney remodeling of the roots confirmed the resorptive nature of the lesions. Given the extent of the lesions, and poor prognosis with conservative treatment alone, teeth affected by the most severe resorption were extracted at each visit using a flap technique combined with alveolar vestibular osteotomy. Dental resorptive lesions are rarely detected in the dog but may be more frequent than previously thought. The routine use of dental radiographs can be used to reveal these lesions in the dog. PMID- 21638266 TI - [Survey 2011]. PMID- 21638267 TI - [Employment termination without notice]. PMID- 21638268 TI - [Fixed costs - you made your bed, now sleep in it]. PMID- 21638269 TI - Recent progress in DNA origami technology. AB - DNA origami is an emerging technology for designing defined two-dimensional DNA nanostructures. In this review, we focus on and describe several types of DNA origami-related studies, as follows: (1) programmed DNA origami assembly, (2) DNA origami-templated molecular assembly, (3) design and construction of various three-dimensional DNA origami structures, (4) programmed functionalization of DNA origami and combination with top-down nanotechnology, (5) single molecular observation on a designed DNA origami, and (6) DNA nanomachines working on a DNA origami. PMID- 21638271 TI - Analysis of multidimensional G-quadruplex melting curves. AB - Multidimensional "3D" melting curves for G-quadruplexes are obtained by recording whole spectra (absorbance, CD, fluorescence) as a function of temperature, rather than the common approach of recording the spectral response to temperature at a single wavelength. 3D melting curves are richer in information, and can be used to enumerate the number of significant species and intermediate states required to properly analyze the thermal denaturation reaction to obtain thermodynamic information. This unit describes the application of the method of singular value decomposition to the analysis of 3D melting data obtained for G-quadruplex structures, and how the results of such an analysis can be used to provide a more complete characterization of the mechanism of quadruplex unfolding. PMID- 21638270 TI - Resolution of quadruplex polymorphism by size-exclusion chromatography. AB - This unit describes a method for separation of quadruplex species formed from the same sequence via size-exclusion chromatography (SEC). Polymorphism is inherent to quadruplex formation, and even relatively simple quadruplex-forming sequences, such as the human telomere sequence d(GGG(TTAGGG)(3)), can form a myriad of possible configurations. HPLC, especially using reversed-phase and anion-exchange methods, has been a mainstay of nucleic acids research and purification for many decades. These methods have been applied for separation of individual quadruplex species formed in a mixture from the same parent sequence. PMID- 21638272 TI - RNA synthesis by reverse direction process: phosphoramidites and high purity RNAs and introduction of ligands, chromophores, and modifications at 3'-end. AB - We have synthesized and studied the coupling properties of 3'-DMT-5'-CE phosphoramidites. The coupling efficiency per step surpasses 99% in the reverse direction synthesis methodology, leading to high-purity RNA in a large number of 20- to 21-mers and long-chain oligonucleotides. Our data show that 5'->3' direction synthesis has a distinct advantage compared to the conventional method. As a result, this method of RNA synthesis is expected to be a very useful and practical method of choice for therapeutic-grade RNA. The phosphoramidites, Rev-A n-bz, Rev-C-n-bz, Rev-C-n-ac, Rev-G-n-ac, and Rev-rU are routinely produced with an HPLC purity of greater than 98% and (31)P NMR purity greater than 99.5%. PMID- 21638273 TI - GATA-3 up-regulation in CD8+ T cells as a biomarker of immune dysfunction in systemic sclerosis, resulting in excessive interleukin-13 production. AB - OBJECTIVE: Despite the importance of interleukin-13 (IL-13) in systemic sclerosis (SSc) and other fibrotic diseases, its mechanisms of action are not understood. We have reported that excessive amounts of IL-13 are produced by peripheral blood effector CD8+ T cells from patients with diffuse cutaneous SSc (dcSSc). The aim of the present study was to establish the molecular basis of IL-13 dysregulation in the pathogenesis of SSc. METHODS: Quantitative polymerase chain reaction analysis and intracellular staining were used to study the transcription factors that control naive peripheral blood CD8+ T cell differentiation into type 1 and type 2 cytokine-secreting cells. Intracellular staining revealed that GATA-3 levels in freshly isolated naive CD8+ T cells correlated with specific clinical manifestations. We therefore assessed the effects of GATA-3 inhibition on IL-13 production in CD8+ T cells from the SSc patients. RESULTS: Freshly isolated naive peripheral blood CD8+ T cells expressed high levels of GATA-3 and failed to down regulate IL-13 production when cultured under type 1-skewing conditions, but maintained adequate levels of interferon-gamma production. Cellular GATA-3 levels were significantly higher in patients with dcSSc and early inflammatory disease. Silencing of GATA-3 with small interfering RNA significantly reduced IL-13 production by CD8+ T cells, demonstrating a causal relationship between GATA-3 and IL-13. CONCLUSION: These results provide important new insights into SSc pathogenesis and suggest that increased GATA-3 expression in CD8+ T cells could be a highly relevant biomarker of immune dysfunction in patients with dcSSc. GATA 3 could be a novel therapeutic target for this currently incurable disease. PMID- 21638274 TI - Sulfonated amphipols: synthesis, properties, and applications. AB - Amphipols (APols) are amphiphatic polymers that keep membrane proteins (MPs) water-soluble. The best characterized and most widely used APol to date, A8-35, comprises a polyacrylate backbone grafted with octyl- and isopropylamine side chains. The nature of its hydrophilic moieties prevents its use at the slightly acidic pH that is desirable to slow down the rate of amide proton exchange in solution NMR studies. We describe here the synthesis and properties of pH insensitive APols obtained by replacing isopropyles with taurine. Sulfonated APols (SAPols) can be used to trap MPs in the form of small complexes, to stabilize them, and to keep them water-soluble even at low pH. [(15) N,(1) H] transverse relaxation-optimized spectroscopy NMR spectra obtained at pH 6.8 of a bacterial outer MP folded in SAPols show that the protein is correctly folded. The spectra have a resolution similar to that achieved with A8-35 and reveal water-exposed amide and indole protons whose resonance peaks are absent at pH 8.0. PMID- 21638275 TI - The intrinsic viscosity of linear DNA. AB - We measured the intrinsic viscosity of very small synthetic DNA molecules, of 20 395 base pairs, and incorporated them in a nearly complete picture for the whole span of molecular weights reported in the literature to date. A major transition is observed at M approximately 2 * 10(6) . It is found that in the range of approximately 7 * 10(3) <= M <= 2 * 10(6) , the intrinsic viscosity scales as [eta] approximately M(1.05) , suggesting that short DNA chains are not as rigid as generally thought. The corresponding scaling for the range of 2 * 10(6) <= M <= 8 * 10(10) is [eta] approximately M(0.69) . A comparison of our results with existing equations, for much narrower data distributions, is made, and the agreement is very satisfactory considering the huge range of data analyzed here. Experimental concerns such as the effect of ionic strength, polydispersity, temperature, and shear rate are discussed in detail. Some issues concerning the Huggins coefficient, polymer chain stiffness, and the relationship between the Mark-Houwink constants K, alpha are also presented; it is found that log K = 1.156 - 6.19alpha. PMID- 21638276 TI - Sentinel lymph node biopsy and survival in elderly patients with cutaneous melanoma. AB - BACKGROUND: Sentinel lymph node biopsy (SNB) is a widely adopted staging procedure in patients with cutaneous melanoma. The benefits of SNB have not been evaluated thoroughly in older age groups. METHODS: This was a two-centre retrospective observational study of patients with melanoma aged at least 70 years undergoing SNB. RESULTS: A total of 423 patients were included. SNB was successful in 405 patients (95.7 per cent), of whom 88 (21.7 per cent) had sentinel node metastasis. During a median follow-up of 2.5 years, recurrence developed in 80 patients (18.9 per cent). Nodal recurrence developed in eight sentinel node-negative patients, giving a false-negative rate of 8.3 per cent, a sensitivity of 91.7 per cent and an overall diagnostic accuracy of 98.0 per cent. A total of 46 patients (10.9 per cent) died from melanoma and 42 (9.9 per cent) from other causes. At 5 years, the relapse-free survival rate was 80.0 per cent in sentinel node-negative patients and 39 per cent in node-positive patients; cancer-specific survival rates were 88.6 per cent and 46 per cent respectively (P < 0.001). In multivariable analysis, sentinel node metastasis (P < 0.001), a Breslow thickness of at least 2.0 mm (P = 0.007) and presence of ulceration (P = 0.012) were independent prognostic factors for cancer-specific survival. CONCLUSION: SNB is a feasible and accurate technique for detecting nodal metastases in older patients with melanoma. Sentinel node status is the most important predictor of cancer-specific outcome in the elderly. PMID- 21638277 TI - Clinical and technical outcomes from a randomized clinical trial of endovenous laser ablation compared with conventional surgery for great saphenous varicose veins. AB - BACKGROUND: This report describes the clinical effectiveness and recurrence rates from a randomized trial of endovenous laser ablation (EVLA) and surgery for varicose veins. METHODS: Some 280 patients were randomized equally using sealed opaque envelopes to two parallel groups: surgery and EVLA. Inclusion criteria included symptomatic disease secondary to primary, unilateral, isolated saphenofemoral junction incompetence, leading to reflux into the great saphenous vein (GSV). Outcomes were: technical success, recurrent varicose veins on clinical examination, patterns of reflux on duplex ultrasound examination, and the effect of recurrence on quality of life, assessed by the Aberdeen Varicose Vein Questionnaire (AVVQ). Assessments were at 1, 6, 12 and 52 weeks after the procedure. RESULTS: Initial technical success was greater following EVLA: 99.3 versus 92.4 per cent (P = 0.005). Surgical failures related mainly to an inability to strip the above-knee GSV. The clinical recurrence rate at 1 year was lower after EVLA: 4.0 versus 20.4 per cent (P < 0.001). The number of patients needed to treat with EVLA rather than surgery to avoid one recurrence at 1 year was 6.3 (95 per cent confidence interval 4.0 to 12.5). Twelve of 23 surgical recurrences were related to an incompetent below-knee GSV and ten to neovascularization. Of five recurrences after EVLA, two were related to neoreflux in the groin tributaries and one to recanalization. Clinical recurrence was associated with worse AVVQ scores (P < 0.001). CONCLUSION: EVLA treatment had lower rates of clinical recurrence than conventional surgery in the short term. PMID- 21638278 TI - Outcomes after adjuvant chemotherapy in the treatment of high-risk urothelial carcinoma of the upper urinary tract (UUT-UC): results from a large multicenter collaborative study. AB - BACKGROUND: Urothelial carcinoma of the upper urinary tract (UUT-UC) was a rare, aggressive urologic cancer with a propensity for multifocality, local recurrence, and metastasis. High-risk patients had poor outcomes. Because of the rarity of these tumors, randomized clinical trials and data regarding adjuvant chemotherapy in locally advanced tumors are currently unavailable. Our objective was to assess the effect of adjuvant chemotherapy and the impact of potential prognostic factors on survival in high-risk, postsurgical UUT-UC patients. METHODS: Using a multi-institutional, international retrospective database, identified were 627 patients with high risk UUT-UCs (pT3N0, pT4N0 and/or N+ and/or M+) who underwent surgical removal. Only patients who received adjuvant chemotherapy were included. RESULTS: Overall, 140 patients (22.6%) with a median age of 67 years were included. The median follow-up was 22.5 months. The 5-year, overall survival for the entire cohort was 43%, the 5-year recurrence-free survival was 54%, and metastasis-free survival was 53% at 5 years. Positive surgical margins were an independent prognostic factor for recurrence (P = .06), cancer-specific mortality (P = .05), and overall mortality (P = .02) of any cause. Adjuvant chemotherapy was not linked with overall or cancer-specific survival in patients with high risk disease (adjuvant chemotherapy [n = 140] vs no treatment [n = 487]) (P >.5). CONCLUSIONS: Adjuvant postoperative chemotherapy did not offer any significant benefit to overall survival in our population. Additional data were necessary, and studies enrolling patients at high risk in clinical trials investigating neoadjuvant chemotherapy in conjunction with chemotherapy should have been highly encouraged. PMID- 21638279 TI - Prognostic impact of severe thrombocytopenia in low-risk myelodysplastic syndrome. AB - BACKGROUND: Thrombocytopenia is very common in myelodysplastic syndrome (MDS); however, its clinical impact in low-risk patients remains controversial. METHODS: The authors analyzed the incidence and prognostic significance of thrombocytopenia at diagnosis in 2565 de novo MDS patients included in the Spanish MDS Registry. RESULTS: Thrombocytopenia (platelet count <100 * 10(9) /L) was identified in 842 patients (32.8%). Severe thrombocytopenia (platelet count <30 * 10(9) /L) was observed in 7.1% of patients and was significantly associated with a higher-risk World Health Organization subtype (P = .026) and intermediate 2/high-risk International Prognostic Scoring System (IPSS) score (P = .046). Severe thrombocytopenia was the most important prognostic factor and had negative effects on the low/intermediate-1 risk group. Median overall survival of patients with a platelet count <30 and >= 30 * 10(9) /L was 16 months and 71 months, respectively (hazard ratio, 4.66; 95% confidence interval, 2.74-7.90; P < .0001). The negative effect of severe thrombocytopenia in low/intermediate-1 risk patients was caused by increased risk of bleeding. CONCLUSIONS: MDS patients with low/intermediate-1 IPSS risk score and severe thrombocytopenia should no longer be regarded as low risk, and must be considered for disease-altering approaches at diagnosis. PMID- 21638280 TI - 2'-5' oligoadenylate synthetase 1 polymorphism is associated with prostate cancer. AB - BACKGROUND: The antiviral, proapoptotic, antiproliferative gene 2'-5' oligoadenylate synthetase (2-5OAS1) converts adenosine triphosphate into a series of 2'-5' oligoadenylates (2-5A). In turn, 2-5A activates latent ribonuclease (RNaseL), a candidate hereditary prostate cancer gene. OAS1 polymorphism (reference single nucleotide polymorphism [SNP] 2660 [rs2660]) has been associated with increased susceptibility to infections and various diseases. In general, the low-enzyme-activity adenine-adenine (AA) genotype promotes susceptibility, whereas the high-enzyme-activity guanosine-guanosine (GG) genotype confers protection. In this study, the authors investigated the association of this functional OAS1 polymorphism (rs2660) with prostate cancer. METHODS: Sample size and power were calculated using a power calculation software program for case-control genetic association analyses. Genomic DNA samples from a control group (n = 140) and from a case group of patients with prostate cancer (n = 164) were used for genotyping SNPs rs2660, rs1131454, and rs34137742 in all samples. Statistical analyses were performed using a logistic regression model. RESULTS: A significant association was observed between the rs2660 genotype (A/G) and prostate cancer. Genotype AA increased the risk, whereas genotype GG decreased the risk of prostate cancer. The GG genotype was not observed in the African American samples. The AA genotype also increased the risk of prostate cancer with age. CONCLUSIONS: The OAS1 SNP rs2660 AA genotype was associated significantly with prostate cancer, whereas the GG genotype protected against prostate cancer. OAS1 rs2660 may be a prostate cancer susceptibility polymorphism, which is a significant observation, especially in a context of the OAS1-RNaseL pathway. Thus, a functional defect in OAS1 because of the rs2660 SNP not only can attenuate RNaseL function but also can alter cell growth and apoptosis independent of RNaseL. PMID- 21638281 TI - Comparison of drug permeabilities across the blood-retinal barrier, blood-aqueous humor barrier, and blood-brain barrier. AB - Drugs vary in their ability to permeate the blood-retinal barrier (BRB), blood aqueous humor barrier (BAB), and blood-brain barrier (BBB) and the factors affecting the drug permeation remain unclear. In this study, the permeability of various substances across BRB, BAB, and BBB in rats was determined using the brain uptake index (BUI), retinal uptake index (RUI), and aqueous humor uptake index (AHUI) methods. Lipophilic substances showed high permeabilities across BBB and BRB. The RUI values of these substances were approximately four-fold higher than the BUI values. The AHUI versus lipophilicity curve had a parabolic shape with AHUI(max) values at log D(7.4) ranging from -1.0 to 0.0. On the basis of the difference on the lipophilicities, verapamil, quinidine, and digoxin showed lower permeability than predicted from those across BBB and BRB, whereas only digoxin showed a lower permeability across BRB. These low permeabilities were significantly increased by P-glycoprotein inhibitors. Furthermore, anion transporter inhibition increased the absorption of digoxin to permeate into the retina and aqueous humor. In conclusion, this study suggests that efflux transport systems play an important role in the ocular absorption of drugs from the circulating blood after systemic administration. PMID- 21638282 TI - Comprehensive quantitative and qualitative liquid chromatography-radioisotope mass spectrometry analysis for safety testing of tolbutamide metabolites without standard samples. AB - Liquid chromatography-radioisotope-mass spectrometry (LC-RI-MS) analysis was used to determine the structures of 12 (four previously unknown) (14) C-tolbutamide (TB) metabolites in rat biological samples (plasma, urine, bile, feces, and microsomes). The four novel metabolites are omega-carboxy TB, hydroxyl TB (HTB)-O glucuronide, TB-ortho or meta-glutathion, and tolylsulphoaminocarbo-glutathion. In rat plasma, after oral administration of (14) C-TB at therapeutic dose (1 mg/kg) and microdose (1.67 ug/kg), the total RI and six metabolites [HTB, carboxy TB (CTB), M1: desbutyl TB, M2: omega-hydroxyl TB, M3: alpha-hydroxyl TB, and M4: omega-1-hydroxyl TB] were quantified by LC-RI-MS. The plasma concentrations were calculated using their response factors (MS-RI intensity ratio) without standard samples, and the area under the curve (AUC) of plasma concentration per time for evaluation of Safety Testing of Drug Metabolites (MIST) was calculated using the ratio of TB metabolites AUC/total RI AUC. The ratios were as follows: TB 94.5% and HTB 2.5% for the microdose (1.67 ug/kg) and TB 95.6%, HTB 0.96%, CTB 0.065%, M1 0.62%, M2 0.0035%, M3 0.077%, and M4 0.015% for the therapeutic dose (1 mg/kg). These values were less than 10% of the MIST criteria. PMID- 21638283 TI - PLGA microdevices for retinoids sustained release produced by supercritical emulsion extraction: continuous versus batch operation layouts. AB - Retinyl acetate (RA) was selected as a model compound to be entrapped in poly(lactic-co-glycolic)acid (PLGA) microspheres using supercritical emulsion extraction (SEE). Several oil-in-water emulsions prepared using acetone and aqueous glycerol (80% glycerol, 20% water) were processed using supercritical carbon dioxide (SC-CO2 ) to extract the oily phase and to induce microspheres formation. The characteristics of the microspheres obtained by conventional liquid emulsion extraction and SEE were also compared: SEE produced spherical and free flowing microspheres, whereas the conventional liquid-liquid extraction showed large intraparticles aggregation. Emulsion extraction by SC-CO2 technology was tested using two different operation layouts: batch (SEE-B) and continuous (SEE-C). SEE-C was performed using a packed tower to produce emulsion/SC-CO2 contact in countercurrent mode, allowing higher microsphere recovery and process efficiencies. Operating at 80 bar and 36 degrees C, SEE-C produced PLGA/RA microspheres with mean sizes between 3.3 and 4.5 MUm with an excellent encapsulation efficiency of 80%-90%. Almost all the drug was released in about 6 days when charged at 2.7% (w/w), whereas only 40% and 10% of RA were released in the same period of time when the charge was 5.2% and 8.8% (w/w), respectively. Release kinetics constants calculated from the experimental data, using a mathematical model, were also proposed and discussed. PMID- 21638284 TI - Genetic variations of orosomucoid genes associated with serum alpha-1-acid glycoprotein level and the pharmacokinetics of paclitaxel in Japanese cancer patients. AB - Alpha-1-acid glycoprotein (AGP) encoded by orosomucoid genes (ORM1 and ORM2) is an acute-phase response protein and functions as a drug-binding protein that affects pharmacokinetics (PK)/pharmacodynamics of binding drugs. To explore the effects of genetic variations of ORMs and a role of AGP on paclitaxel (PTX) therapy, we analyzed the duplication and genetic variations/haplotypes of ORMs in 165 Japanese cancer patients and then investigated their associations with serum AGP levels and the PK parameters of PTX. No effects of ORM duplications on serum AGP levels at baseline or PK of PTX were observed, but close associations of ORM1 -559T > A with the increases of AGP levels and area under the curve (AUC) of PTX metabolites were detected. In addition, a significant correlation between the serum AGP level and the AUCs of PTX metabolites was observed, suggesting that AGP may function as a carrier of PTX from the blood into the liver via putative receptors. This study provided useful information on the possible clinical importance of ORM genetic polymorphisms and a novel role of AGP in PTX therapy. PMID- 21638285 TI - Opalescence of an IgG2 monoclonal antibody solution as it relates to liquid liquid phase separation. AB - Opalescence for a monoclonal antibody solution was systematically studied with respect to temperature, protein concentration, ionic strength (using KCl), and pH conditions. Multiple techniques, including measurement of light scattering at 90 degrees and transmission, Tyndall test, and microscopy, were deployed to examine the opalescence behavior. Near the vicinity of the critical point on the liquid liquid coexistence curve in the temperature-protein concentration phase diagram, the enhanced concentration fluctuations significantly contributed to the critical opalescence evidently by formation of small liquid droplets. Furthermore, our data confirm that away from the critical point, the opalescence behavior is related to the antibody self-association (agglomeration) caused by the attractive antibody-antibody interactions. As expected, at a pH near the pI of the antibody, the solution became less opalescent as the ionic strength increased. However, at a pH below the pI, the opalescence of the solution became stronger, reached a maximum, and then began to drop as the ionic strength further increased. The change in the opalescence correlated well with the trends of protein-protein interactions revealed by the critical temperature from the liquid-liquid phase separation. PMID- 21638286 TI - Nationwide surveillance of 18 respiratory viruses in patients with influenza-like illnesses: a pilot feasibility study in the French Sentinel Network. AB - The aim of the present study was to test the feasibility of integrating the diagnosis of 18 respiratory viruses into clinical surveillance of influenza-like illness using a PCR-DNA microarray detection assay. The study took place in the French Sentinel Network, a nationwide surveillance network of General Practitioners (GPs) representative of French GPs in terms age, location, and type of practice (urban/rural). Three virological laboratories also participated in the study. The study was planned for 5 weeks from January 25, 2010 to February 27, 2010. A subset of 150 Sentinel GPs, located in mainland France, was enrolled to collect clinical data and nasopharyngeal samples from every first patient of the week having a medical visit for influenza-like illness defined as a sudden fever of 39 degrees C or more with respiratory symptoms and myalgia. Sixty-three GPs (42%) collected 103 samples while 87 GPs (58%) did not. GPs did not differ with respect to their age, gender, urban/rural distribution, or years of inscription in the Sentinel Network. Patients included were of a similar age and had similar vaccination characteristics, but were more frequently men than influenza-like illness patients reported to the network during the study period. Sixty-one viruses were detected from 56 of 96 (58%) interpretable samples. The respiratory viruses detected most frequently were metapneumovirus and respiratory syncytial virus. This study showed that virological diagnosis of 18 respiratory viruses can be combined with surveillance of clinical influenza-like illness in general practice. Although feasibility has not been demonstrated yet, it will be evaluated over the winter of 2010-2011. PMID- 21638287 TI - Effect of cardiac resynchronization therapy on libido and erectile dysfunction. AB - BACKGROUND: Chronic heart failure (HF) is a common, complex clinical syndrome characterized by dyspnea, fatigue and exercise intolerance. HF patients experience decreased libido and erectile dysfunction (ED). The effects of cardiac resynchronization therapy (CRT) on libido and erectile function have not been previously evaluated. We aimed to investigate the effects of CRT on libido and ED. HYPOTHESIS: Cardiac resynchronization therapy improves libido and ED. METHODS: Thirty-one male patients with advanced HF, scheduled for implantation of a CRT device, were included in the study. Left ventricular systolic function, New York Heart Association (NYHA) class, libido, and ED were assessed before and 6 months after CRT. Libido and ED were evaluated with the Aging Male Symptoms (AMS) rating scale and internationally validated Sexual Health Inventory for Men (SHIM) questionnaire, respectively. RESULTS: At the 6-month follow-up, the mean NYHA class improved from 3.4 +/- 0.5 to 2.1 +/- 0.6 (P<0.001). On echocardiographic examination, an improvement in left ventricular ejection fraction (LVEF) from 18 +/- 5% to 32 +/- 6% was detected (P<0.001). A significant increase in mean SHIM score and a significant decrease in mean AMS were noted. Changes in SHIM and AMS scores were correlated positively with the increase in LVEF (r = 0.47, P = 0.007 and r = - 0.36, P = 0.04, respectively). Similarly, SHIM scores were correlated negatively (r = - 0.57, P = 0.001) and AMS scores were correlated positively (r = 0.73, P = 0.0001) with the improvement in NYHA class. CONCLUSIONS: CRT results in a significant improvement in libido and erectile function in patients with congestive HF. This improvement is related to the improvements in the LVEF and functional capacity. . PMID- 21638288 TI - A new approach for semiconductor parameter extraction using cathodoluminescence and artificial neural networks. AB - In this paper, a new parameter extraction technique that jointly extracts four semiconductor-related parameters from theoretical/experimental cathodoluminescence data collected as a function of electron-beam energy is presented. The extraction technique is based on feed-forward artificial neural networks (ANN) where the ANN is trained to learn the inherent relationship between the input parameters (absorption coefficient alpha, diffusion length L, dead layer thickness Zt, and relative quantum efficiency Q) and the output parameter (CL intensity versus electron beam energy). After the training of the ANN, it is possible to observe the reverse process and extract the four parameters from any CL curve using an exhaustive search method. One of the main advantages of the proposed method is that the optimum set of values for the four parameters (alpha, L, Zt, Q) are obtained because the exhaustive search is performed in the search space spanned by all four parameters. Computational results on an n-type GaAs free defect semiconductor sample show that a unique set of parameter values with errors less than 5.5% from the nominal values can be obtained for each set of the experimental data points using the proposed algorithm. PMID- 21638289 TI - Can X-ray spectrum imaging replace backscattered electrons for compositional contrast in the scanning electron microscope? AB - The high throughput of the silicon drift detector energy dispersive X-ray spectrometer (SDD-EDS) enables X-ray spectrum imaging (XSI) in the scanning electron microscope to be performed in frame times of 10-100 s, the typical time needed to record a high-quality backscattered electron (BSE) image. These short duration XSIs can reveal all elements, except H, He, and Li, present as major constituents, defined as 0.1 mass fraction (10 wt%) or higher, as well as minor constituents in the range 0.01-0.1 mass fraction, depending on the particular composition and possible interferences. Although BSEs have a greater abundance by a factor of 100 compared with characteristic X-rays, the strong compositional contrast in element-specific X-ray maps enables XSI mapping to compete with BSE imaging to reveal compositional features. Differences in the fraction of the interaction volume sampled by the BSE and X-ray signals lead to more delocalization of the X-ray signal at abrupt compositional boundaries, resulting in poorer spatial resolution. Improved resolution in X-ray elemental maps occurs for the case of a small feature composed of intermediate to high atomic number elements embedded in a matrix of lower atomic number elements. XSI imaging strongly complements BSE imaging, and the SDD-EDS technology enables an efficient combined BSE-XSI measurement strategy that maximizes the compositional information. If 10 s or more are available for the measurement of an area of interest, the analyst should always record the combined BSE-XSI information to gain the advantages of both measures of compositional contrast. PMID- 21638290 TI - Developments in low-voltage microscopy instrumentation. AB - Low-voltage microscopy can produce high-resolution, high-contrast images of nanometer-scale surface features. Several techniques exist for low-energy electron generation, but most successful in current instrumentation is the combination of a magnetic lens with an electrostatic immersion lens. This compound lens allows the low-voltage column to maintain high potential in the column while independently controlling the landing energy on the sample. Aberration coefficients decrease linearly with the immersion ratio leading to larger optimum semiconvergent angle on the sample and improvement in spot size and beam current. Most recently highly scaled miniature electron beam columns have shown promising low-voltage performance from all electrostatic lenses operating only at low potential. This article explores the current design of low voltage columns, including variations on compound lenses and miniature lenses, and from a system level, discusses the strength and weaknesses of each design. PMID- 21638291 TI - Atomic force microscopy demonstration of cytoskeleton instability in mouse erythrocytes with dematin-headpiece and beta-adducin deficiency. AB - The pattern of disassembly of the cytoskeletal network of murine erythrocytes with deficiency of either dematin-headpiece or beta-adducin or both proteins were investigated using atomic force microscopy. A heterogeneous complex structure with fine filament features and coarse features was observed in the cytoskeleton of wild type mouse erythrocytes, whereas a significant amount of rearrangement and aggregation occurred in the mutants, particularly in the cells carrying double gene mutations. These results are consistent with the cellular and biochemical phenotype of the mutant cell membranes as being more fragile due to weakened vertical connections with the plasma membrane. PMID- 21638292 TI - A novel graphical representation of protein sequences and its application. AB - On the basis of information on the evolution of the 20 amino acids and their physiochemical characteristics, we propose a new two-dimensional (2D) graphical representation of protein sequences in this article. By this representation method, we use 2D data to represent three-dimensional information constructed by the amino acids' evolution index, the class information of amino acid based on physiochemical characteristics, and the order of the amino acids appearing in the protein sequences. Then, using discrete Fourier transform, the sequence signals with different lengths can be transformed to the frequency domain, in which the sequences are with the same length. A new method is used to analyze the protein sequence similarity and to predict the protein structural class. The experiments indicate that our method is effective and useful. PMID- 21638293 TI - Single electron densities: a new tool to analyze molecular wavefunctions. AB - A new partitioning scheme for the electron density of a many-electron wavefunction into single electron densities is proposed. These densities are based on the most probable arrangement of the electrons in an atom or molecule. Therefore, they contain information about the electron-electron interaction and, most notably, the Fermi hole due to the antisymmetry of the many-electron wavefunction. The single electron densities overlap and can be combined to electron pair distributions close to the qualitative electron pairs that represent, for instance, the basis of the valence shell electron pair repulsion model. Single electron analyses are presented for the water, ethane, and ethene molecules. The effect of electron correlation on the single electron and pair densities is investigated for the water molecule. PMID- 21638294 TI - Theoretical modulation of the color of light emitted by firefly oxyluciferin. AB - One of the major mysteries regarding firefly bioluminescence is its pH-dependent multicolor variation. At basic pH, the emission is on the yellow-green region, whereas at acid pH, the light emission is observed on the red region of the visible spectrum. Theoretical calculations using density functional theory, molecular mechanics, and semiempirical methods were made to investigate the effect exerted by intermolecular forces on light emission, and their modulation by polarity, and the differences in the conformation of the active site at basic and acid pH. Red emission is achieved by the weakening of the interactions of the emitter with ionic and hydrophobic molecules, by the polarization of the benzothiazole microenvironment, by ionization of the enzyme-emitter complex and by changes of the hydrogen bond network. Arg220, Glu346, Ala350, Leu344 and adenosine-5'-monophosphate have blue-shifting effects, while His247, Phe249, Gly341, Thr253, and Ile288 exert a redshifting one. PMID- 21638295 TI - Brownian dynamics simulations on CPU and GPU with BD_BOX. AB - There has been growing interest in simulating biological processes under in vivo conditions due to recent advances in experimental techniques dedicated to study single particle behavior in crowded environments. We have developed a software package, BD_BOX, for multiscale Brownian dynamics simulations. BD_BOX can simulate either single molecules or multicomponent systems of diverse, interacting molecular species using flexible, coarse-grained bead models. BD_BOX is written in C and employs modern computer architectures and technologies; these include MPI for distributed-memory architectures, OpenMP for shared-memory platforms, NVIDIA CUDA framework for GPGPU, and SSE vectorization for CPU. PMID- 21638296 TI - Prenatal pharmacotherapy for fetal anomalies: a 2011 update. AB - Fetal therapy can be defined as any prenatal treatment administered to the mother with the primary indication to improve perinatal or long-term outcomes for the fetus or newborn. This review provides an update of the pharmacological therapies that are solely directed at the fetus with anomalies and outlines a future transcriptomic approach. Fetal anomalies targeted with prenatal pharmacotherapy are a heterogeneous group of structural, endocrine, and metabolic conditions, including congenital cystic adenomatoid malformation (CCAM), congenital adrenal hyperplasia, congenital heart block, fetal tachyarrhythmias, inborn errors of metabolism, fetal thyroid disorders, and polyhydramnios. To date, the majority of pharmacotherapies for fetal anomalies have been evaluated only in retrospective, uncontrolled studies. The way forward will be with an evidence-based approach to prenatal pharmacological interventions. PMID- 21638297 TI - The effect of L-arginine or L-citrulline supplementation on biochemical parameters and the vascular aortic wall in high-fat and high-cholesterol-fed rats. AB - The aim of the present study is to investigate the potential role of L-arginine or L-citrulline in rats fed high-fat and high-cholesterol (HFC) diet. HFC feeding increased significantly serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities, urea and all lipid profiles and decreased significantly serum high-density lipoprotein-cholesterol (HDL-c) and non significantly serum nitric oxide levels. L-arginine or L-citrulline administration reversed the increase in serum AST and ALT activities, urea and all lipid profiles. These effects were associated with a concomitant increase in HDL-c and nitric oxide levels. In general, rats fed HFC diet and orally treated with L-arginine or L-citrulline had higher relative percentage of 18:0, 20:0 and 22:6 and lower 16:0 fatty acids than rats fed HFC diet. Light and transmission electron microscopic findings of the thoracic aorta confirmed the biochemical results and demonstrated structural changes in the endothelial cells of the intimal layer, medial smooth muscle cells as well as in the adventitial layer in HFC fed-animals. However, these findings indicate little structural alterations in animals supplemented with L-arginine or L-citrulline along with HFC feeding. In the present study, L-arginine or L-citrulline was effective hypocholesterolemic and hypolipidemic agents in rats. PMID- 21638298 TI - Effects of erythropoietin on ICAM-1 and PECAM-1 expressions on human umbilical vein endothelial cells subjected to oxidative stress. AB - The protective effect of erythropoietin (Epo) is based on its ability to reduce oxidation and to stabilize the cells. The aim of the study was to evaluate the influence of Epo on malonyl dialdehyde (MDA), intercellular adhesion molecule-1 (ICAM-1) (CD54) and platelet-endothelial cell adhesion molecule-1 (PECAM-1) (CD31) levels on human umbilical vein endothelial cells (HUVECs) stimulated by tumour necrosis factor-alpha (TNF-alpha). HUVECs were incubated with Epo (10-40 IU ml-1) or TNF-alpha (10-40 ng ml-1) alone or preincubated with Epo (20 IU ml-1) and subsequently stimulated with TNF-alpha (10-40 ng ml-1). MDA concentrations were measured using the high-performance liquid chromatography, whereas ICAM-1 and PECAM-1 expressions were evaluated by flow cytometry. Incubation with Epo resulted in a decrease in MDA and the increased expressions of ICAM-1 and PECAM 1. Exposure to TNF-alpha reflected an increase in MDA, ICAM-1 and PECAM-1 levels. These changes were inhibited by preincubation with Epo. The cytoprotective activity proven in this study points to new applications and therapeutic possibilities for Epo. PMID- 21638299 TI - The reduction of SIRT1 in livers of old mice leads to impaired body homeostasis and to inhibition of liver proliferation. AB - Age declines liver functions, leading to the development of age-associated diseases. A member of the sirtuins family, SIRT1, is involved in the control of glucose homeostasis and fat metabolism. Because aging livers have alterations in glucose and fat metabolism, we examined a possible role of SIRT1 in these alterations. We found that aged livers have a reduced expression of SIRT1 and have lost proper control of the regulation of SIRT1 after partial hepatectomy (PH). Down-regulation of SIRT1 in the liver of old mice is mediated by CCAAT/Enhancer Binding Protein/histone deacetylase 1 (C/EBPbeta-HDAC1) complexes, which bind to and repress the SIRT1 promoter. In the livers of young mice, SIRT1 is activated after PH and supports high levels of glucose and triglycerides during liver regeneration. In old mice, however, C/EBPbeta-HDAC1-mediated repression of the SIRT1 promoter blocks activation of SIRT1, leading to low levels of glucose and triglycerides during liver regeneration. Down-regulation of SIRT1 in the livers of young mice resulted in alterations similar to those observed in the livers of old mice, whereas the normalization of SIRT1 in the livers of old mice corrects the levels of glucose and triglycerides after PH. The normalization of SIRT1 in old mice also improves liver regeneration via the elimination of the C/EBPalpha-Brm complex. These studies showed a critical role of the reduction of SIRT1 in age-associated liver dysfunctions and provide a potential tool for the correction of liver functions in old patients after surgical resections. PMID- 21638300 TI - Constrained S-estimators for linear mixed effects models with covariance components. AB - Linear mixed effects (LME) models are increasingly used for analyses of biological and biomedical data. When the multivariate normal assumption is not adequate for an LME model, then a robust estimation approach is preferable to the maximum likelihood one. M-estimators were considered before for robust estimation of the LME models, and recently a constrained S-estimator was proposed. This S estimator cannot be applied directly to LME models with correlated error terms and vector random effects with correlated dimensions. Therefore, a modification is proposed, which extends application of the constrained S-estimator to the LME models for multivariate responses with correlated dimensions and to longitudinal data. Also, a new computational algorithm is developed for computing constrained S-estimators. Performance of the S-estimators based on the original Tukey's biweight and translated biweight is evaluated in a small simulation study with repeated multivariate responses with correlated dimensions. The proposed methodology is applied to jointly analyze repeated measures on three cholesterol components, HDL, LDL, and triglycerides. PMID- 21638301 TI - Assessment of a disease screener by hierarchical all-subset selection using area under the receiver operating characteristic curves. AB - In many clinical settings, a commonly encountered problem is to assess the accuracy of a screening test for early detection of a disease. In this article, we develop hierarchical all-subset variable selection methods to assess and improve a psychosis screening test designed to detect psychotic patients in primary care clinics. We select items from an existing screener to achieve best prediction accuracy based on a gold standard psychosis status diagnosis. The existing screener has a hierarchical structure: the questions fall into five domains, and there is a root question followed by several stem questions in each domain. The statistical question lies in how to implement the hierarchical structure in the screening items when performing variable selection such that when a stem question is selected in the screener, its root question should also be selected. We develop an all-subset variable selection procedure that takes into account the hierarchical structure in a questionnaire. By enforcing a hierarchical rule, we reduce the dimensionality of the search space, thereby allowing for fast all-subset selection, which is usually computationally prohibitive. To focus on prediction performance of a selected model, we use area under the ROC curve as the criterion to rank all admissible models. We compare the procedure to a logistic regression-based approach and a stepwise regression that ignores the hierarchical structure. We use the procedure to construct a psychosis screening test to be used at a primary care clinic that will optimally screen low-income, Latino psychotic patients for further specialty referral. PMID- 21638302 TI - Metabolism of thalidomide by human liver microsome cytochrome CYP2C19 is required for its antimyeloma and antiangiogenic activities in vitro. AB - In this study, we used a system of human liver microsomes to investigate the antimyeloma and antiangiogenic activities of thalidomide. Myeloma cells and human umbilical vein endothelial cells (HUVECs) were treated with thalidomide alone or thalidomide incubated with human liver microsomal protein. We found that thalidomide alone had no direct effect on several multiple myeloma cell lines (U266, NCI-H929, RPMI 8226, LP-1, CZ-1) or on HUVECs in vitro. However, when incubated with human liver microsomal protein, thalidomide (100 ug/ml) caused a decrease of 34.9-46.7% in cell viability in myeloma cells and 12% in HUVECs. Cell cycle analysis and apoptosis detection indicated that the decreases in cell viability were correlated with the induction of apoptosis. Thalidomide incubated with microsomal protein also influenced HUVEC migration and tube formation. These effects were partially reversed by omeprazole (10 umol/l), a potent inhibitor of CYP2C19, suggesting that CYP2C19 is required for thalidomide to exhibit its antimyeloma and antiangiogenic activities. PMID- 21638303 TI - Clinical implications of acute myeloid leukemia presenting as myeloid sarcoma. AB - In this retrospective study, we aim to analyze the characteristics, treatments, and overall survival of all patients presenting with isolated myeloid sarcoma (MS) or MS with concomitant acute myeloid leukemia (AML) compared with all patients with AML, treated during the same period. We identified patients with AML with or without MS at diagnosis, presenting to our medical center between the years 1990 and 2005. There was no statistically significant difference between the groups regarding gender, age, cytogenetic risk groups, rate of complete remission, number of cycles of chemotherapy needed to achieve complete remission, and rate of first relapse. The time to death in the MS group was not significantly different (p = 0.60) from the AML group, and radiotherapy did not affect the median time to death. Transplantation prolonged survival in both groups (p = 0.018 and p < 0.0001, respectively). Patients with MS at diagnosis might benefit from upfront aggressive treatment with hematopoietic stem cell transplantation. PMID- 21638304 TI - Cellular (FLICE) like inhibitory protein (cFLIP) expression in diffuse large B cell lymphoma identifies a poor prognostic subset, but fails to predict the molecular subtype. AB - Immunohistochemistry can sub-classify diffuse large B-cell lymphoma (DLBCL) into germinal centre B-cell like (GCB) and non-GCB subtypes. The latter consists predominately of the activated B-cell like subgroup in which nuclear factor kappa B activation is its characteristic. Expression of cellular caspase 8 (FLICE)-like inhibitory protein (cFLIP), a caspase 8 homologue, is regulated by nuclear factor kappa-B signalling, and it is the main inhibitor of Fas ligand activated apoptosis. To determine if cFLIP expression was confined to non-GCB subtype, we studied 66 cases of DLBCL. cFLIP expression showed no significant correlation to DLBCL subtypes (GCB or non-GCB) but was associated with a worse clinical outcome. For cFLIP positive and negative patients, the five-year event free survival was 20 and 31%, respectively (p = 0.049), and the five-year overall survival was 20 and 57%, respectively (p = 0.041). PMID- 21638305 TI - Phosphorylation status of heat shock protein 27 influences neurite growth in adult dorsal root ganglion sensory neurons in vitro. AB - The small heat shock protein Hsp27 influences neurite growth, potentially via phosphorylation-dependent interactions of Hsp27 with actin. To investigate the contribution of Hsp27 phosphorylation to neurite growth in adult DRG neurons, we employed hamster Hsp27 cDNA constructs (in pIRES-EGFP) with mutations in the phosphorylation sites, either mimicking constitutively phosphorylated Hsp27 (with substitution of serines 15 and/or 90 by glutamate) or preventing phosphorylation at the site (serines 15/90 replaced by alanine). Five mutant constructs were employed in this study in addition to wild-type hamster Hsp27; siRNA directed against the rat Hsp27 was used to depress endogenous Hsp27. Neurite growth was assessed in EGFP-expressing cells following immunocytochemistry and tracing of neurite growth. Hsp27 staining and phalloidin labelling were used to examine Hsp27 and actin colocalization in neurons and growth cones. Overall, our results demonstrate that the role that Hsp27 plays in neurite growth can be affected by phosphorylation, oligomerization, or a combination of both. Hsp27 constructs that are able to dimerize and/or form large oligomers [WT, Hsp27-AA, Hsp27-AE, Hsp27 Delta(5-23)] rescued siRNA-depressed neurite growth, whereas Hsp27 mutants that do not form dimers or oligomers (Hsp27-EE and Hsp27-EA) were unable to rescue the effect of the siRNA. The phalloidin labelling qualitatively showed a higher level of localization of actin with the Hsp27-AA compared with the other constructs. Although phosphorylation appears to be important in growth, the ability of Hsp27 to exist in both phospho- and nonphospho- states is likely key to its role in regulating cytoskeletal elements involved in neurite growth. PMID- 21638306 TI - Feeding mechanics and dietary implications in the fossil sloth Neocnus (Mammalia: Xenarthra: Megalonychidae) from Haiti. AB - Haitian species of the extinct ground sloth genus Neocnus (Mammalia: Pilosa: Megalonychidae) have previously been hypothesized to have a much reduced jugal bone and a correspondingly reduced masseter musculature but a paucity of specimens has prevented further investigation of this hypothesis. Recent discovery of jugal bones belonging to Haitian specimens of Neocnus within the University of Florida Museum collections enables the element to be more accurately described. The discovery also makes it possible to explore mastication in these sloths. Osteological characters related to feeding were examined, along with comparative estimations of bite force with the extant tree sloths, Bradypus and Choloepus, and their known dietary habits as a means to infer aspects of the paleodiet of Neocnus. There is a significant difference in moment arm calculations for m. masseter between predicted and actual jugals, but the overall significance for bite force is lost and hampered by small sample size. Neocnus demonstrates a variety of characters that are similar to those of Bradypus and not to Choloepus, which is a close phylogenetic relative. The masticatory musculature of Neocnus enabled a chewing cycle emphasizing a grinding combination of mesiodistal and linguobuccal movements of the molariform dentition. The orientations of m. masseter and m. temporalis are estimated to produce relatively high bite force ratios that imply a masticatory system with stronger versus faster components. Because of the similarity of bite forces and jaw mechanics to those of Bradypus, in addition to a number of osteological adaptations indicative of herbivorous grazers (elevated mandibular condyle, large and complex masseter, and robust angular process), the Haitian forms of Neocnus are considered to have been selective feeders with a folivorous diet. PMID- 21638307 TI - Parasitization by Macrocentrus cingulum (Hymenoptera: Braconidae) influences expression of prophenoloxidase in Asian corn borer Ostrinia furnacalis. AB - A prophenoloxidase (PPO) cDNA (OfPPO) was cloned from the Asian corn borer Ostrinia furnacalis. Sequence analysis revealed a full length transcript of the OfPPO cDNA with 2,686 bp, containing a 2,079 bp open reading frame (ORF), a 73-bp 5'-untranslated region, and a 534-bp 3'-untranslated region with a poly(A) signal. The ORF encodes a 693-amino acid polypeptide, containing two distinct copper-binding regions, a plausible thiol ester site, two proteolytic activation sites, and a conserved C-terminal region, but lacks a signal peptide sequence. Expression of the OfPPO transcript in the plasma, hemocytes, fat body and midgut was inhibited by Macrocentrus cingulum at 4 h post-parasitization (pp). In situ hybridization analysis showed that the hemocytes, especially the oenocytoids, hybridized strongly with the DNA probes of the OfPPO gene. No signal was detected in the cuticular epithelium or fat body of the parasitized larvae. Colloidal gold particles were used to visualize the PPO by immunoelectron microscopy. The time course study revealed a decrease in the labeling of the OfPPO at 4, 6, 8, 12, and 1 day pp in the larval integument and midgut parasitized by M. cingulum. We infer from time course studies of OfPPO gene expression and PO enzymatic activity that OfPPO in the integument is released from hemocytes and that the OfPPO expression was influenced at the transcriptional, translational, and then the post translational level by parasitization challenge. PMID- 21638308 TI - Characterization of two closely related alpha-amylase paralogs in the bark beetle, Ips typographus (L.). AB - Ips typographus (L.), the eight-spined spruce bark beetle, causes severe damage throughout Eurasian spruce forests and suitable nuclear markers are needed in order to study its population structure on a genetic level. Two closely related genes encoding alpha-amylase in I. typographus were characterized and named AmyA and AmyB. Both alpha-amylase paralogs consisted of six exons and five introns. AmyA encodes a polypeptide of 483 amino acids, whereas AmyB has two alternative transcripts encoding polypeptides of 483 and 370 amino acids. The expression levels of both genes were high during larval stage and adulthood. The AmyB transcripts were absent in the pupal stage. A modification of the allozyme staining method allowed us to detect two clusters of bands on the electrophoretic gel that may correspond to the two alpha-amylase genes. There was a correlation between the lack of AmyB expression in pupa and the absence of the fast migrating isozyme cluster at this stage, suggesting that the faster migrating isoforms are products of the AmyB gene, whereas the slowly migrating bands are derived from the AmyA. PMID- 21638309 TI - Dissecting prenatal, postnatal, and inherited effects: ART and design. AB - With the failure of common variants alone to explain the bulk of trait heritability, it becomes more important to understand the contribution of maternally inherited effects, prenatal effects, and postnatal environmental effects. These effects can be disentangled by studying families containing children conceived by assisted reproductive technologies (ART). We propose and develop a model that is an extension of the variance component model commonly used in pedigree analysis. Our model is flexible enough to allow any number of family members and degrees of relationship; thus, researchers can use both small and extended families simultaneously. Simulations demonstrate that our method has appropriate statistical properties and is robust to model misspecification and accurate in the presence of missing data. Most importantly, our method is able to disentangle maternally inherited effects from prenatal effects, which are confounded in traditional family studies. Our analyses also provide guidance to researchers designing studies that will use ART families to clarify genetic and environmental factors underlying traits. PMID- 21638310 TI - The road to recovery: Egypt's healthcare reform. AB - As many industrial and third-world countries recover from the severe economic crisis of a global recession, they continue to struggle with its negative effect on their healthcare systems. Healthcare reform has become a leading policy agenda item for most countries. This is especially true for countries in the developing world who are struggling to allocate very limited resources to meet the growing health needs of their residents and the expectations of global health. In the late 1990s, the Egyptian government, in conjunction with the United States Agency for International Development, initiated a Health Sector Reform Program (HSRP) to completely reform the way healthcare was financed, organized and delivered with the intent to extend healthcare coverage to all of its citizens. Although some successes have resulted from the HSRP, Egypt's new government leaders will need to be informed on policies that may more effectively improve the health of the Egyptian population. PMID- 21638311 TI - Comparing the cost of community pharmacy and mail-order pharmacy in a US retirement system. AB - Although community pharmacies have been the mainstay for drug distribution in the USA, plan members are encouraged to use mail-order pharmacies as a cost containment strategy. Both channels differ with respect to reimbursement rates, utilization, and costs. We evaluated the differences in reimbursement rates and in ingredient costs between the two dispensing channels. We used pharmacy claims from a large Midwestern retirement system for the period 2000-2005. A representative sample of drug products was selected. We estimated the aggregated gross reimbursement, the ingredient cost, dispensing fee, pharmacy incentives for drug substitution, professional fee for other services, sales tax, and reimbursement per payer. The sample contained 1964 observations-four million claims. There were 58.5% observations for single source brands and 39.0% for generics. Observations with lower unit gross reimbursement rate in community pharmacy increased from 10.3% to 16.5%. Unit ingredient cost and dispensing fees were higher in community pharmacy than in mail-order pharmacy. Community pharmacy had a lower reimbursement rate per unit of medication (33.5-44.6% observations) compared with mail-order pharmacy. There were 87.3-98.1% observations with a higher patient co-financing per unit of medication in community pharmacy. Gross pharmaceutical reimbursement rates and unit ingredient costs were higher in community pharmacy than in mail-order pharmacy; but in more than 10% of the observations, the costs were higher in mail-order pharmacy than in community pharmacy. PMID- 21638312 TI - Understanding the impact of conflict on health services in Iraq: information from 401 Iraqi refugee doctors in Jordan. AB - OBJECTIVES: This study aims to assess how conflict has affected the function of Iraqi health services and its doctors. METHODS: Interviews were conducted in person or by mobile phone with 401 Iraqi doctors entering Jordan since 2003, using respondent-driven sampling methods. RESULTS: Of the Iraqi doctors interviewed in 2008, 94% came from Baghdad, although 25% had moved within Iraq in the past year. They reported a steady year-by-year decline in Iraqi health services from 2003 through 2006, with perhaps some improvement in 2007. By 2006, 67% of doctors said essential drugs were present less than half of the time (95% confidence interval [CI] 54-81), and 69% (95% CI 56-84) said essential equipment was available or working half the time or less. By 2006, 95% said their facilities lacked skilled health workers, and 90% noted reduced quality of care. Violent death rates among doctors in Baghdad reached 47.6/1000/yr (95% CI 42.0 53.7) in 2006. In the same year, migration rates for Baghdad doctors moving elsewhere in Iraq were 143.8/1000/yr (95% CI 134.0-154.1), and departure from Iraq was 299.5/1000/yr (95% CI 285.3-314.3). CONCLUSIONS: Deterioration of health services quality, staffing levels and violence against doctors continued from 2003 through 2006, although these may have improved slightly in 2007. In 2009 and 2010, reports suggest that assassinations of doctors and out-migration have continued. Few have returned. PMID- 21638313 TI - Are public health authorities able to "steer" rather than "row"? An empirical analysis in the Italian National Health Service. AB - The nature of the local health authorities (LHAs) in the Italian National Health Service has been deeply reformed during the 1990s by new public management (NPM) reforms that introduced decentralization, quasi-market and managerialism. These reforms implied that the main role of LHA is to govern the production of health services in their area (steer) rather than to only directly produce services (row). After more than 15 years from these reforms of Italian healthcare, we describe how much the steering versus rowing dichotomy made an impact on LHA activity, through an analysis of the management control systems they set up for themselves and the subsequent qualitative analysis of the opinions that a diverse group of managers expressed during 8 days of group discussion. Results show that managers of Italian LHAs, when only a small part of services is produced, tend to perceive their steering role as impossible to play and focus on production, leaving therefore ungoverned a significant part of the services offered to residents. NPM, therefore, was able to influence the reform of Italian healthcare but, as suggested by a postmodernist interpretation, left managers with a rhetoric change based on inconsistent assumptions instead of actionable ideas to manage the change process. PMID- 21638314 TI - A multilevel model of patient safety culture: cross-level relationship between organizational culture and patient safety behavior in Taiwan's hospitals. AB - BACKGROUND: As health-care organizations endeavor to improve their quality of care, there is a growing recognition of the importance of establishing a culture of patient safety. The main objective of this study was to investigate the cross level influences of organizational culture on patient safety behavior in Taiwan's hospitals. METHODS: The authors measured organizational culture (bureaucratic, supportive and innovative culture), patient safety culture and behavior from 788 hospital workers among 42 hospitals in Taiwan. Multilevel analysis was applied to explore the relationship between organizational culture (group level) and patient safety behavior (individual level). RESULTS: Patient safety culture had positive impact on patient safety behavior in Taiwan's hospitals. The results also indicated that bureaucratic, innovative and supportive organizational cultures all had direct influence on patient safety behavior. However, only supportive culture demonstrated significant moderation effect on the relationship between patient safety culture and patient safety behavior. Furthermore, organizational culture strength was shown correlated negatively with patient safety culture variability. CONCLUSIONS: Overall, organizational culture plays an important role in patient safety activities. Safety behaviors of hospital staff are partly influenced by the prevailing cultural norms in their organizations and work groups. For management implications, constructed patient priority from management commitment to leadership is necessary. For academic implications, research on patient safety should consider leadership, group dynamics and organizational learning. These factors are important for understanding the barriers and the possibilities embedded in patient safety. PMID- 21638315 TI - A novel quercetin analogue from a medicinal plant promotes peak bone mass achievement and bone healing after injury and exerts an anabolic effect on osteoporotic bone: the role of aryl hydrocarbon receptor as a mediator of osteogenic action. AB - We recently reported that extracts made from the stem bark of Ulmus wallichiana promoted peak bone mass achievement in growing rats and preserved trabecular bone mass and cortical bone strength in ovariectomized (OVX) rats. Further, 6-C-beta-D glucopyranosyl-(2S,3S)-(+)-3',4',5,7-tetrahydroxyflavanol (GTDF), a novel flavonol-C-glucoside isolated from the extracts, had a nonestrogenic bone-sparing effect on OVX rats. Here we studied the effects of GTDF on osteoblast function and its mode of action and in vivo osteogenic effect. GTDF stimulated osteoblast proliferation, survival, and differentiation but had no effect on osteoclastic or adipocytic differentiation. In cultured osteoblasts, GTDF transactivated the aryl hydrocarbon receptor (AhR). Activation of AhR mediated the stimulatory effect of GTDF on osteoblast proliferation and differentiation. Furthermore, GTDF stimulated cAMP production, which mediated osteogenic gene expression. GTDF treatments given to 1- to 2-day-old rats or adult rats increased the mRNA levels of AhR target genes in calvaria or bone marrow stromal cells. In growing female rats, GTDF promoted parameters of peak bone accrual in the appendicular skeleton, including increased longitudinal growth, bone mineral density, bone-formation rate (BFR), cortical deposition, and bone strength. GTDF promoted the process of providing newly generated bone to fill drill holes in the femurs of both estrogen sufficient and -deficient rats. In osteopenic OVX rats, GTDF increased BFR and significantly restored trabecular bone compared with the ovaries-intact group. Together our data suggest that GTDF stimulates osteoblast growth and differentiation via the AhR and promotes modeling-directed bone accrual, accelerates bone healing after injury, and exerts anabolic effects on osteopenic rats likely by a direct stimulatory effect on osteoprogenitors. Based on these preclinical data, clinical evaluation of GTDF as a potential bone anabolic agent is warranted. PMID- 21638316 TI - GPM6B regulates osteoblast function and induction of mineralization by controlling cytoskeleton and matrix vesicle release. AB - Neuronal membrane glycoprotein gene (GPM6B) encodes a membrane glycoprotein that belongs to the proteolipid protein family. We identified GPM6B as a gene that is strongly upregulated during osteoblast differentiation. To investigate the role of GPM6B in the process of bone formation, we silenced GPM6B expression during osteogenic differentiation of human mesenchymal stem cells (hMSCs). GPM6B silencing in hMSCs resulted in reduced alkaline phosphate (ALP) activity along with reduced mineralization of extracellular matrix (ECM). Microarray expression analysis of GPM6B-depleted osteogenic hMSCs revealed significant changes in genes involved in cytoskeleton organization and biogenesis. Immunocytochemistry results confirm changes in the distribution of actin filaments, as well as the shape and size of focal adhesions on GPM6B silencing. Moreover, we demonstrated that production and release of ALP-positive matrix vesicles (MVs) were reduced. In conclusion, we identified GPM6B as a novel regulator of osteoblast function and bone formation. This finding demonstrates the significance of cytoskeleton organization for MV production and subsequent mineralization. PMID- 21638317 TI - Identification of quantitative trait loci influencing skeletal architecture in mice: emergence of Cdh11 as a primary candidate gene regulating femoral morphology. AB - Bone strength is influenced by many properties intrinsic to bone, including its mass, geometry, and mineralization. To further advance our understanding of the genetic basis of bone-strength-related traits, we used a large (n = 815), moderately (G(4) ) advanced intercross line (AIL) of mice derived from a high runner selection line (HR) and the C57BL/6J inbred strain. In total, 16 quantitative trait loci (QTLs) were identified that affected areal bone mineral density (aBMD) and femoral length and width. Four significant (p < .05) and one suggestive (p < .10) QTLs were identified for three aBMD measurements: total body, vertebral, and femoral. A QTL on chromosome (Chr.) 3 influenced all three aBMD measures, whereas the other four QTLs were unique to a single measure. A total of 10 significant and one suggestive QTLs were identified for femoral length (FL) and two measures of femoral width, anteroposterior (AP) and mediolateral (ML). FL QTLs were distinct from loci affecting AP and ML width, and of the 7 AP QTLs, only three affected ML. A QTL on Chr. 8 that explained 7.1% and 4.0% of the variance in AP and ML, respectively, was mapped to a 6-Mb region harboring 12 protein-coding genes. The pattern of haplotype diversity across the QTL region and expression profiles of QTL genes suggested that of the 12, cadherin 11 (Cdh11) was most likely the causal gene. These findings, when combined with existing data from gene knockouts, identify Cdh11 as a strong candidate gene within which genetic variation may affect bone morphology. PMID- 21638318 TI - Association of the alpha(2)delta(1) subunit with Ca(v)3.2 enhances membrane expression and regulates mechanically induced ATP release in MLO-Y4 osteocytes. AB - Voltage-sensitive calcium channels (VSCCs) mediate signaling events in bone cells in response to mechanical loading. Osteoblasts predominantly express L-type VSCCs composed of the alpha(1) pore-forming subunit and several auxiliary subunits. Osteocytes, in contrast, express T-type VSCCs and a relatively small amount of L type alpha(1) subunits. Auxiliary VSCC subunits have several functions, including modulating gating kinetics, trafficking of the channel, and phosphorylation events. The influence of the alpha(2)delta auxiliary subunit on T-type VSCCs and the physiologic consequences of that association are incompletely understood and have yet to be investigated in bone. In this study we postulated that the auxiliary alpha(2) delta subunit of the VSCC complex modulates mechanically regulated ATP release in osteocytes via its association with the T-type Ca(v) 3.2 (alpha(1H) ) subunit. We demonstrated by reverse-transcriptase polymerase chain reaction, Western blotting, and immunostaining that MLO-Y4 osteocyte-like cells express the T-type Ca(v)3.2(alpha(1H)) subunit more abundantly than the L-type Ca(v)1.2 (alpha(1C)) subunit. We also demonstrated that the alpha(2) delta(1) subunit, previously described as an L-type auxiliary subunit, complexes with the T-type Ca(v)3.2 (alpha(1H)) subunit in MLO-Y4 cells. Interestingly, siRNA mediated knockdown of alpha(2) delta(1) completely abrogated ATP release in response to membrane stretch in MLO-Y4 cells. Additionally, knockdown of the alpha(2)delta(1) subunit resulted in reduced ERK1/2 activation. Together these data demonstrate a functional VSCC complex. Immunocytochemistry following alpha(2)delta(1) knockdown showed decreased membrane localization of Ca(v) 3.2 (alpha(1H)) at the plasma membrane, suggesting that the diminished ATP release and ERK1/2 activation in response to membrane stretch resulted from a lack of Ca(v) 3.2 (alpha(1H)) at the cell membrane. PMID- 21638319 TI - A single infusion of zoledronic acid produces sustained remissions in Paget disease: data to 6.5 years. AB - Two trials have shown that a single 5-mg infusion of zoledronic acid achieves much higher response rates in Paget disease of bone than risedronate. The duration of this effect is unknown. We have conducted an open follow-up of responders from the two trials (152 originally treated with zoledronic acid, 115 with risedronate) out to 6.5 years without further intervention. Endpoints were times to relapse (ie, return of serum total alkaline phosphatase activity to within 20% of the pretreatment value) or loss of response (response = normalization of alkaline phosphatase or 75% or greater reduction in its excess). Bone turnover markers were lower in the zoledronic acid group throughout follow up, with mean alkaline phosphatase (ALP) remaining within the reference range in these patients, whereas the mean in the risedronate group was above normal from 1 year. Relapse rates were substantially greater in the risedronate group (23 of 115, 20%) than in those treated with zoledronic acid (1 of 152, 0.7%, p < .001), and loss of response occurred in 19 (12.5%) zoledronic acid patients compared with 71 (62%) risedronate patients (p < .0001). Risk ratios for relapse and loss of response in zoledronic acid patients were 0.02 [95% confidence interval (CI) 0.00-0.18] and 0.12 (95% CI 0.07-0.19), respectively. Changes from baseline in quality of life, assessed using SF-36 scores, were more positive in the zoledronic acid group across the follow-up period (p = .01). Bone markers at 6 months were predictive of response duration. These data demonstrate an unprecedented duration of remission of Paget disease following treatment with zoledronic acid, accompanied by an improved quality of life. PMID- 21638320 TI - High levels of beta-catenin signaling reduce osteogenic differentiation of stem cells in inflammatory microenvironments through inhibition of the noncanonical Wnt pathway. AB - Periodontal ligament stem cells (PDLSCs), a new population of mesenchymal stem cells (MSCs), have been isolated from the periodontal ligament (PDL). The capacity of multipotency and self-renewal makes them an excellent cell source for bone regeneration and repair. However, their bone-regeneration ability could be awakened in inflammatory microenvironments, which may be the result of changes in their differentiation potential. Recently, genetic evidences has shown that the Wnt pathway plays an important role in bone homeostasis. In this study we have determined the specific role of beta-catenin in osteogenic differentiation of PDLSCs obtained from inflammatory microenvironments (P-PDLSCs). The inflammatory microenvironment, while inhibiting osteogenic differentiation potential, promotes proliferation of MSCs. A higher the level of beta-catenin in P-PDLSCs than in H PDLSCs (PDLSCs obtained from a healthy microenvironment) resulted in the same disparity in canonical Wnt signaling pathway activation between each cell type. Here we show that activation of beta-catenin suppresses the noncanonical Wnt/Ca(2+) pathway, leading to increased proliferation but reduced osteogenic differentiation of P-PDLSCs. Downregulation of the levels of beta-catenin by treatment with dickkopf-1 (DKK-1) leads to activation of the noncanonical Wnt/Ca(2+) pathway, which, in turn, results in the promotion of osteogenic differentiation in P-PDLSCs. Interestingly, beta-catenin can affect both the canonical Wnt/beta-catenin pathway and the noncanonical Wnt/Ca(2+) pathway. Our data indicate that beta-catenin plays a central role in regulating osteogenic differentiation of MSCs in inflammatory microenvironments. Given the important role of Wnt signaling in osteogenic differentiation, it is possible that agents that can modify this pathway may be of value in bone regeneration by MSCs in chronic inflammatory microenvironments. PMID- 21638321 TI - An essential role for the association of CD47 to SHPS-1 in skeletal remodeling. AB - Integrin-associated protein (IAP/CD47) has been implicated in macrophage macrophage fusion. To understand the actions of CD47 on skeletal remodeling, we compared Cd47(-/-) mice with Cd47(+/+) controls. Cd47(-/-) mice weighed less and had decreased areal bone mineral density compared with controls. Cd47(-/-) femurs were shorter in length with thinner cortices and exhibited lower trabecular bone volume owing to decreased trabecular number and thickness. Histomorphometry revealed reduced bone-formation and mineral apposition rates, accompanied by decreased osteoblast numbers. No differences in osteoclast number were observed despite a nonsignificant but 40% decrease in eroded surface/bone surface in Cd47( /-) mice. In vitro, the number of functional osteoclasts formed by differentiating Cd47(-/-) bone marrow cells was significantly decreased compared with wild-type cultures and was associated with a decrease in bone-resorption capacity. Furthermore, by disrupting the CD47-SHPS-1 association, we found that osteoclastogenesis was markedly impaired. Assays for markers of osteoclast maturation suggested that the defect was at the point of fusion and not differentiation and was associated with a lack of SHPS-1 phosphorylation, SHP-1 phosphatase recruitment, and subsequent dephosphorylation of non-muscle cell myosin IIA. We also demonstrated a significant decrease in osteoblastogenesis in bone marrow stromal cells derived from Cd47(-/-) mice. Our finding of cell autonomous defects in Cd47(-/-) osteoblast and osteoclast differentiation coupled with the pronounced skeletal phenotype of Cd47(-/-) mice support the conclusion that CD47 plays an important role in regulating skeletal acquisition and maintenance through its actions on both bone formation and bone resorption. PMID- 21638322 TI - Differential role of dopamine in emotional attention and memory: evidence from Parkinson's disease. AB - Consistent with the hypothesis that dopamine is implicated in the processing of salient stimuli relevant to the modification of various behavioral responses, Parkinson's disease is associated with emotional blunting. To address the hypothesis that emotional attention and memory are modulated by dopaminergic neurotransmission in Parkinson's disease, we assessed 15 nondemented patients with Parkinson's disease while on and off dopaminergic medication and 15 age matched healthy controls. Visual stimuli were presented, and recognition was used to assess emotional memory. Response latency was used as a measure of emotional attention modulation. Stimuli were varied based on valence (pleasant, neutral, and unpleasant) and arousal (high and low) dimensions. Controls had significantly better memory for positive than negative stimuli, whereas patients with Parkinson's disease tested off medication had significantly better memory for negative than positive items. This negativity bias was lost when they were tested while on dopaminergic medication. Reaction times in patients with Parkinson's disease off medication were longer than in healthy controls and, paradoxically, were even longer when on medication. Further, although both healthy controls and patients with Parkinson's disease in the "off" state had arousal-induced prolongation of reaction time, this effect was not seen in patients with Parkinson's disease on medication. These data indicate that dopaminergic neurotransmission is implicated in emotional memory and attention and suggest that dopamine mediates emotional memory via the valence dimension and emotional attention via arousal. Furthermore, our findings suggest that emotional changes in Parkinson's disease result from the effects of both the disease process and dopaminergic treatment. PMID- 21638323 TI - No evidence for THAP1/DYT6 variants as disease modifiers in DYT1 dystonia. PMID- 21638324 TI - A within-subject comparison of 6-[18F]fluoro-m-tyrosine and 6-[18F]fluoro-L-dopa in Parkinson's disease. AB - Progression of Parkinson's disease symptoms is imperfectly correlated with positron emission tomography biomarkers for dopamine biosynthetic pathways. The radiopharmaceutical 6-[(18) F]fluoro-m-tyrosine is not a substrate for catechol-O methyltransferase and therefore has a more favorable uptake-to-background ratio than 6-[(18) F]fluoro-L-dopa. The objective of this study was to evaluate 6-[(18) F]fluoro-m-tyrosine relative to 6-[(18) F]fluoro-L-dopa with partial catechol-O methyltransferase inhibition as a biomarker for clinical status in Parkinson's disease. Twelve patients with early-stage Parkinson's disease, off medication, underwent Unified Parkinson Disease Rating Scale scoring, brain magnetic resonance imaging, and 3-dimensional dynamic positron emission tomography using equivalent doses of 6-[(18) F]fluoro-m-tyrosine and 6-[(18) F]fluoro-L-dopa with tolcapone, a catechol-O-methyltransferase inhibitor. Images were realigned within subject, after which the tissue-derived uptake rate constant was generated for volumes of interest encompassing the caudate nucleus, putamen, and subregions of the putamen. We computed both bivariate (Pearson) and partial (covariate of age) correlations between clinical subscores and tissue-derived uptake rate constant. Tissue-derived uptake rate constant values were correlated between the radiopharmaceuticals (r = 0.8). Motor subscores were inversely correlated with the contralateral putamen 6-[(18) F]fluoro-m-tyrosine tissue-derived uptake rate constant (|r| > 0.72, P < .005) but not significantly with the 6-[(18) F]fluoro-L dopa tissue-derived uptake rate constant. The uptake rate constants for both radiopharmaceuticals were also inversely correlated with activities of daily living subscores, but the magnitude of correlation coefficients was greater for 6 [(18) F]fluoro-m-tyrosine. In this design, 6-[(18) F]fluoro-m-tyrosine uptake better reflected clinical status than did 6-[(18) F]fluoro-L-dopa uptake. We attribute this finding to 6-[(18) F]fluoro-m-tyrosine's higher affinity for the target, L-aromatic amino acid decarboxylase, and the absence of other major determinants of the uptake rate constant. These results also imply that L aromatic amino acid decarboxylase activity is a major determinant of clinical status. PMID- 21638325 TI - Sensory perception changes induced by transcranial magnetic stimulation over the primary somatosensory cortex in Parkinson's disease. AB - Sensory symptoms are common nonmotor manifestations of Parkinson's disease. It has been hypothesized that abnormal central processing of sensory signals occurs in Parkinson's disease and is related to dopaminergic treatment. The objective of this study was to investigate the alterations in sensory perception induced by transcranial magnetic stimulation of the primary somatosensory cortex in patients with Parkinson's disease and the modulatory effects of dopaminergic treatment. Fourteen patients with Parkinson's disease with and without dopaminergic treatment and 13 control subjects were included. Twenty milliseconds after peripheral electrical tactile stimuli in the contralateral thumb, paired-pulse transcranial magnetic stimulation over the right primary somatosensory cortex was delivered. We evaluated the perception of peripheral electrical tactile stimuli at 2 conditioning stimulus intensities, set at 70% and 90% of the right resting motor threshold, using different interstimulus intervals. At 70% of the resting motor threshold, paired-pulse transcranial magnetic stimulation over the right primary somatosensory cortex induced an increase in positive responses at short interstimulus intervals (1-7 ms) in controls but not in patients with dopaminergic treatment. At 90% of the resting motor threshold, controls and patients showed similar transcranial magnetic stimulation effects. Changes in peripheral electrical tactile stimuli perception after paired-pulse transcranial magnetic stimulation over the primary somatosensory cortex are altered in patients with Parkinson's disease with dopaminergic treatment compared with controls. These findings suggest that primary somatosensory cortex excitability could be involved in changes in somatosensory integration in Parkinson's disease with dopaminergic treatment. PMID- 21638326 TI - Odor identification deficits identify Parkinson's disease patients with poor cognitive performance. AB - Olfactory dysfunction is a prodromal and prevalent nonmotor symptom of Parkinson's disease. Unlike olfactory dysfunction in Alzheimer's disease, it is believed to be unrelated to cognitive impairment. However, recent research has implicated cholinergic denervation in Parkinson's disease hyposmia and linked it to verbal memory. This research hypothesized that severe odor identification deficits may identify patients with Parkinson's disease at risk for cognitive impairment. The current study tested this hypothesis by comparing 24 functionally anosmic, nondemented patients with Parkinson's disease and 39 nonanosmic, nondemented patients with Parkinson's disease with 29 healthy control participants on composite scores of memory, processing speed, executive function, and language. The functionally anosmic group had significantly poorer visual and verbal memory than the nonanosmic group, which was indistinguishable from the control group. Furthermore, the functionally anosmic group had reduced processing speed compared with the nonanosmic patients with Parkinson's disease, who, in turn, were outperformed by the control group. On the composite language score, the score of the functionally anosmic group was significantly reduced compared with that of the control group, whereas the nonanosmic group scored in the medium range. The 2 patient groups did not differ on executive functioning. These findings demonstrate co-occurrence between reduced cognitive function and olfactory deficits in functionally anosmic patients with Parkinson's disease and support the notion of more severe cognitive deficits in this group. PMID- 21638327 TI - Association between plasma paroxetine concentration and changes in plasma brain derived neurotrophic factor levels in patients with major depressive disorder. AB - Recent studies have implicated brain-derived neurotrophic factor (BDNF) in the pathophysiology of depression and in the activities of antidepressant drugs. Serum BDNF levels are lower in depressed patients and increase in response to antidepressant medications; however, no studies have examined the association between plasma concentrations of antidepressant drugs and plasma BDNF levels. We assessed plasma BDNF levels and paroxetine concentrations in 45 patients with major depression who were being treated with paroxetine. Plasma samples were collected between 10:00 h and 12:00 h at baseline and after 1, 2 and 6 weeks of treatment. The BDNF level and paroxetine concentration of each sample were measured via enzyme immunoassay and high-performance liquid chromatography, respectively. Plasma BDNF levels increased after 2 and 6 weeks of paroxetine treatment. Plasma BDNF levels were significantly lower in men than in women. Changes in plasma BDNF level were correlated with plasma drug concentration after 2 (r = 0.309, p < 0.05) and 6 weeks (r = 0.329, p < 0.05) but not correlated with plasma drug concentration after 1 week (r = 0.284, ns). Multiple regression analysis confirmed that this change was only significantly correlated with plasma paroxetine concentration after 2 (standardised beta = 0.343, p < 0.05) and 6 weeks (standardised beta = 0.375, p < 0.05). These results suggest that paroxetine treatment increases plasma BDNF levels and that plasma paroxetine levels play an important role in changes in plasma BDNF levels. PMID- 21638328 TI - Olanzapine augmentation of milnacipran for stage 2 treatment-resistant major depression: an open study. AB - OBJECTIVE: Olanzapine augmentation of fluoxetine, a selective serotonin reuptake inhibitor, is an effective augmentation therapy for treatment-resistant depression (TRD). However, studies of olanzapine augmentation of other antidepressants are few. We investigated the efficacy and safety of olanzapine augmentation of milnacipran, a serotonin-norepinephrine reuptake inhibitor, for TRD. METHODS: This study covered patients with stage 2 TRD, defined by Thase and Rush. Olanzapine was added to milnacipran, and its dosage was adjusted according to each patient. Previous treatments were continued, but no new treatments were allowed. Response was measured using Hamilton Depression Rating Scale (HAMD) and Clinical Global Impression at weeks 0, 1, 2, 3, 4, and 8. RESULTS: Eleven patients aged 53.2 +/- 24.0 years received olanzapine at 5.0 +/- 1.9 mg/day with milnacipran. HAMD and Clinical Global Impression scores improved significantly from baseline to endpoint. This improvement occurred in week 1. At endpoint, seven of 11 (64%) were responders on HAMD (>= 50% reduction). Four patients (36%) discontinued the trial because of no efficacy. No severe adverse effect occurred. CONCLUSIONS: Olanzapine augmentation of milnacipran for stage 2 TRD might be effective and well tolerated. However, our study is open label and uncontrolled. Therefore, a double-blind controlled trial is necessary to confirm our results. PMID- 21638329 TI - Early improvement on antipsychotic treatment as a predictor of subsequent response in schizophrenia: analyses from ziprasidone clinical studies. AB - OBJECTIVE: We examine data from short-term placebo-controlled and comparator controlled clinical trials of ziprasidone in schizophrenia to confirm the predictive capacity of early symptom changes for response. We pose the question of how early is too early to consider "stay or switch" and evaluate the predictive capability of a clinical measure in this regard. METHODS: We presented two separate pooled analyses of (i) two placebo-controlled and (ii) two active comparator (risperidone and olanzapine) randomized trials of ziprasidone in schizophrenia. Relationship between early changes in Positive and Negative Syndrome Scale (PANSS) total, Brief Psychiatric Rating Scale (BPRS), and Clinical Global Impression-Improvement (CGI-I) scores and treatment outcome was evaluated. RESULTS: Week 2 improvement was more reliably predictive of subsequent outcome than week 1 improvement using PANSS and BPRS scores with high sensitivity and specificity, whereas CGI-I had much lower specificity. Overall, non-improvement at week 1 or week 2 was highly predictive of non-response using BPRS scores and PANSS but not CGI-I. CONCLUSIONS: These data, independent of antipsychotic used, confirm prior research showing that early improvement in symptoms is predictive of response. There appears to be an important window of time, beyond week 1, during which important clinical decisions to stay or switch medication may be made. PMID- 21638330 TI - Association between participant-identified problems and depression severity in problem-solving therapy for low-income homebound older adults. AB - OBJECTIVES: The purpose of this study was to examine the relationship between the severity of baseline depressive symptoms and the problems that low-income homebound older adults (n = 66) identified in their problem-solving therapy (PST) sessions. METHODS: Depressive symptoms were measured with the 24-item Hamilton Rating Scale for Depression (HAMD). Participant-identified problems recorded in the therapists' worksheets were coded into seven categories: living arrangement/housing issues, financial/health care expense issues, family or other relationship issues, hygiene/task issues, social isolation issues, physical/functional health issues, and mental/emotional health issues. T-tests and ordinary least squares regression analyses were used to examine differences in HAMD scores between those who identified any problem in each category and those who did not. RESULTS: Participants who had living arrangement/housing and family or other relationship issues had higher baseline HAMD scores than the rest of the participants. At 2-week posttest, those with living arrangement/housing issues continued to have higher HAMD scores than the others, whereas those with family or other relationship issues did not. CONCLUSION: The study findings provide insights into the problems that low-income, depressed homebound individuals bring to their PST sessions. It was not clear if family conflict or other relationship issues contributed to their depression or vice versa, but it appears that PST may have contributed to alleviating depressive symptoms associated with these issues. Precarious living/housing situations appeared to have had a serious depressogenic effect and could not be easily resolved within a short time frame of the PST process, as these issues required formal support. PMID- 21638331 TI - Incorporation of evolutionary information into Rosetta comparative modeling. AB - Prediction of protein structures from sequences is a fundamental problem in computational biology. Algorithms that attempt to predict a structure from sequence primarily use two sources of information. The first source is physical in nature: proteins fold into their lowest energy state. Given an energy function that describes the interactions governing folding, a method for constructing models of protein structures, and the amino acid sequence of a protein of interest, the structure prediction problem becomes a search for the lowest energy structure. Evolution provides an orthogonal source of information: proteins of similar sequences have similar structure, and therefore proteins of known structure can guide modeling. The relatively successful Rosetta approach takes advantage of the first, but not the second source of information during model optimization. Following the classic work by Andrej Sali and colleagues, we develop a probabilistic approach to derive spatial restraints from proteins of known structure using advances in alignment technology and the growth in the number of structures in the Protein Data Bank. These restraints define a region of conformational space that is high-probability, given the template information, and we incorporate them into Rosetta's comparative modeling protocol. The combined approach performs considerably better on a benchmark based on previous CASP experiments. Incorporating evolutionary information into Rosetta is analogous to incorporating sparse experimental data: in both cases, the additional information eliminates large regions of conformational space and increases the probability that energy-based refinement will hone in on the deep energy minimum at the native state. PMID- 21638332 TI - Protein-protein interactions: analysis of a false positive GST pulldown result. AB - One of the most common ways to demonstrate a direct protein-protein interaction in vitro is the glutathione-S-transferse (GST)-pulldown. Here we report the detailed characterization of a putative interaction between two transcription factor proteins, GATA-1 and Kruppel-like factor 3 (KLF3/BKLF) that show robust interactions in GST-pulldown experiments. Attempts to map the interaction interface of GATA-1 on KLF3 using a mutagenic screening approach did not yield a contiguous binding face on KLF3, suggesting that the interaction might be non specific. NMR experiments showed that the proteins do not interact at protein concentrations of 50-100 MUM. Rather, the GST tag can cause part of KLF3 to misfold. In addition to misfolding, the fact that both proteins are DNA-binding domains appears to introduce binding artifacts (possibly nucleic acid bridging) that cannot be resolved by the addition of nucleases or ethidium bromide (EtBr). This study emphasizes the need for caution in relying on GST-pulldown results and related methods, without convincing confirmation from different approaches. PMID- 21638333 TI - Structural studies of the Nudix GDP-mannose hydrolase from E. coli reveals a new motif for mannose recognition. AB - The Nudix hydrolase superfamily, characterized by the presence of the signature sequence GX(5)EX(7)REUXEEXGU (where U is I, L, or V), is a well-studied family in which relations have been established between primary sequence and substrate specificity for many members. For example, enzymes that hydrolyze the diphosphate linkage of ADP-ribose are characterized by having a proline 15 amino acids C terminal of the Nudix signature sequence. GDPMK is a Nudix enzyme that conserves this characteristic proline but uses GDP-mannose as the preferred substrate. By investigating the structure of the GDPMK alone, bound to magnesium, and bound to substrate, the structural basis for this divergent substrate specificity and a new rule was identified by which ADP-ribose pyrophosphatases can be distinguished from purine-DP-mannose pyrophosphatases from primary sequence alone. Kinetic and mutagenesis studies showed that GDPMK hydrolysis does not rely on a single glutamate as the catalytic base. Instead, catalysis is dependent on residues that coordinate the magnesium ions and residues that position the substrate properly for catalysis. GDPMK was thought to play a role in biofilm formation because of its upregulation in response to RcsC signaling; however, GDPMK knockout strains show no defect in their capacity of forming biofilms. PMID- 21638334 TI - Efficient mapping of ligand migration channel networks in dynamic proteins. AB - For many proteins such as myoglobin, the binding site lies in the interior, and there is no obvious route from the exterior to the binding site in the average structure. Although computer simulations for a limited number of proteins have found some transiently open channels, it is not clear if there exist more channels elsewhere or how the channels are regulated. A systematic approach that can map out the whole ligand migration channel network is lacking. Ligand migration in a dynamic protein resembles closely a well-studied problem in robotics, namely, the navigation of a mobile robot in a dynamic environment. In this work, we present a novel robotic motion planning inspired approach that can map the ligand migration channel network in a dynamic protein. The method combines an efficient spatial mapping of protein inner space with a temporal exploration of protein structural heterogeneity, which is represented by a structure ensemble. The spatial mapping of each conformation in the ensemble produces a partial map of protein inner cavities and their inter-connectivity. These maps are then merged to form a super map that contains all the channels that open dynamically. Results on the pathways in myoglobin for gaseous ligands demonstrate the efficiency of our approach in mapping the ligand migration channel networks. The results, obtained in a significantly less amount of time than trajectory-based approaches, are in agreement with previous simulation results. Additionally, the method clearly illustrates how and what conformational changes open or close a channel. PMID- 21638335 TI - Influence of kinetics of drug binding on EGFR signaling: a comparative study of three EGFR signaling pathway models. AB - We used three models of the epidermal growth factor receptor (EGFR) signaling pathway mimicking three different cell lines to study the effects of kinetics of drug binding on influencing molecular signaling in the pathways. With no incubation of drugs before the external cue epidermal growth factor (EGF) was applied, we found that fast kinetics of binding to protein kinases was advantageous in suppressing the production of the Extracellular signal-regulated kinase (ERK) that triggers cell proliferation, with some exceptions. Incubation of a drug with a protein kinase target for an hour before a pathway was initiated with an external cue made kinetics less significant, so did high concentration of drugs. In addition, we found that applying a drug to a protein kinase mostly affected downstream signaling although upstream events were also affected in a few cases. In examining whether applying two drugs to two protein kinase targets in the pathways could produce synergistic effects, we found positive, negative, or no effects, depending on the protein kinases targeted and the pathway model considered. PMID- 21638336 TI - The role of connexin-36 gap junctions in alcohol intoxication and consumption. AB - Ventral tegmental area (VTA) GABA neurons appear to be critical substrates underlying the acute and chronic effects of ethanol on dopamine (DA) neurotransmission in the mesocorticolimbic system implicated in alcohol reward. The aim of this study was to examine the role of midbrain connexin-36 (Cx36) gap junctions (GJs) in ethanol intoxication and consumption. Using behavioral, molecular, and electrophysiological methods, we compared the effects of ethanol in mature Cx36 knockout (KO) mice and age-matched wild-type (WT) controls. Compared to WT mice, Cx36 KO mice exhibited significantly more ethanol-induced motor impairment in the open field test, but less disruption in motor coordination in the rotarod paradigm. Cx36 KO mice, and WT mice treated with the Cx36 antagonist mefloquine (MFQ), consumed significantly less ethanol than their WT controls in the drink-in-the-dark procedure. The firing rate of VTA GABA neurons in WT mice was inhibited by ethanol with an IC50 of 0.25 g/kg, while VTA GABA neurons in KO mice were significantly less sensitive to ethanol. Dopamine neuron GABA-mediated sIPSC frequency was reduced by ethanol (30 mM) in WT mice, but not affected in KO mice. Cx36 KO mice evinced a significant up-regulation in DAT and D2 receptors in the VTA, as assessed by quantitative RT-PCR. These findings demonstrate the behavioral relevance of Cx36 GJ-mediated electrical coupling between GABA neurons in mature animals, and suggest that loss of coupling between VTA GABA neurons results in disinhibition of DA neurons, a hyper DAergic state and lowered hedonic valence for ethanol consumption. PMID- 21638337 TI - Changes in dynorphin immunoreactivity but unaltered density of enkephalin immunoreactive neurons in basal ganglia nuclei of genetically dystonic hamsters. AB - Dystonia is regarded as a basal ganglia disorder. In the dt(sz) hamster, a genetic animal model of paroxysmal dystonia, previous studies demonstrated a reduced density of striatal GABAergic interneurons which inhibit striatal GABAergic projection neurons. Although the disinhibition of striatal GABAergic projection neurons was evidenced in the dt(sz) hamster, alterations in their density have not been elucidated so far. Therefore, in the present study, the density of striatal methionin-(met-) enkephalin (ENK) immunoreactive GABAergic neurons, which project to the globus pallidus (indirect pathway), was determined in dt(sz) and control hamsters to clarify a possible role of an altered ratio between striatal interneurons and projection neurons. Furthermore, the immunoreactivity of dynorphin A (DYN), which is expressed in entopeduncular fibers of striatal neurons of the direct pathway, was verified by gray level measurements to illuminate the functional relevance of an enhanced striato entopeduncular neuronal activity previously found in dt(sz) hamsters. While the density of striatal ENK immunoreactive (ENK(+) ) neurons did not significantly differ between mutant and control hamsters, there was a significantly enhanced ratio between the DYN immunoreactive area and the whole area of the EPN in dt(sz) hamsters compared to controls. These results support the hypothesis that a disbalance between a reduced density of striatal interneurons and an unchanged density of striatal projection neurons causes imbalances in the basal ganglia network. The consequentially enhanced striato-entopeduncular inhibition leads to an already evidenced reduced activity and an altered firing pattern of entopeduncular neurons in the dt(sz) hamster. PMID- 21638338 TI - Connecting the dots of the cerebro-cerebellar role in cognitive function: neuronal pathways for cerebellar modulation of dopamine release in the prefrontal cortex. AB - Cerebellar involvement in autism, schizophrenia, and other cognitive disorders is typically associated with prefrontal cortical pathology. However, the underlying neuronal mechanisms are largely unknown. It has previously been shown in mice that stimulation of the dentate nucleus (DN) of the cerebellum evokes dopamine (DA) release in the medial prefrontal cortex (mPFC). Here, we investigated the neuronal circuitry by which the cerebellum modulates mPFC DA release. Fixed potential amperometry was used to determine the contribution of two candidate pathways by which the cerebellum may modulate mPFC DA release. In urethane anesthetized mice, DA release evoked by DN stimulation (50 Hz) was recorded in mPFC following local anesthetic lidocaine (0.02 MUg) or ionotropic glutamate receptor antagonist kynurenate (0.5 MUg) infusions into the mediodorsal or ventrolateral thalamic nucleus (ThN md; ThN vl), or the ventral tegmental area (VTA). Following intra-VTA lidocaine or kynurenate infusions, DA release was decreased by ~50%. Following intra-ThN md and ThN vl infusions of either drug, DA release was decreased by ~35% and 15%, respectively. Reductions in DA release following lidocaine or kynurenate infusions were not significantly different indicating that neuronal cells in the VTA and ThN were activated primarily if not entirely by glutamatergic inputs. The present study suggests that neuropathological changes in the cerebellum commonly observed in autism, schizophrenia, and other cognitive disorders could result in a loss of functionality of cerebellar-mPFC circuitry that is manifested as aberrant dopaminergic activity in the mPFC. Additionally, these results specifically implicate glutamate as a modulator of mPFC dopaminergic activity. PMID- 21638339 TI - Expression pattern of Mical-1 in the temporal neocortex of patients with intractable temporal epilepsy and pilocarpine-induced rat model. AB - Mical-1 is a novel F-actin-disassembly factor that is critical in actin reorganization. It provides a molecular conduit through which actin reorganizes-a hallmark of cell morphological changes, including axon navigation. However, whether Mical-1 is involved in the epileptogenesis remains unknown. Here, we investigate Mical-1 expression pattern in patients with intractable temporal lobe epilepsy (TLE) and pilocarpine-induced rat model. We used double-labeled immunoflurescence, immunohistochemistry, and Western blotting to assess the location and expression of Mical-1 in temporal neocortex of patients with intractable TLE, and the expression pattern of Mical-1 at different time point in the hippocampus and temporal lobe cortex of the pilocarpine-induced rat model. Double-labeled immunofluorescence showed that Mical-1 was coexpressed with neuron specific enolase (NSE) in the cytoplasm of neurons in temporal neocortex of patients with TLE and hippocampus of rat model. Faint and scattered immunoreactivity for Mical-1 in the neuron of temporal neocortex in TLE group, but strong immunoreactivity for Mical-1 was shown in control subjects. To quantitatively evaluate the Mical-1 immunoreactivity, we measured the mean optical density (OD) of Mical-1. In the hippocampus of pilocarpine-induced rat model, the OD values transient increased at 6 h after seizure then decreased from 1 day to 14 days, and returned to a subnormal level at 60 days. The lowest level of Mical-1 expression occurred at 14 days after seizure in the hippocampus. In the temporal lobe cortex of rat model, the OD values decreased at all time point after kindling compared to the normal group. Furthermore, our Western blot analysis confirmed these expression patterns of Mical-1 from latent stage to chronic stage. Our results indicate that in patients with TLE and pilocarpine induced rat model, the expression of Mical-1 were followed a downtrend from the latent stage to chronic stage after seizure evoke. Thus, as an effect factor participated in F-actin disassemble, Mical-1 may associate with inner pathophysiological modulation in epilepsy. PMID- 21638341 TI - Attenuating the endoplasmic reticulum stress response improves functional recovery after spinal cord injury. AB - Activation of the unfolded protein response (UPR) is involved in the pathogenesis of numerous CNS myelin abnormalities; yet, its direct role in traumatic spinal cord injury (SCI)-induced demyelination is not known. The UPR is an evolutionarily conserved cell defense mechanism initiated to restore endoplasmic reticulum homeostasis in response to various cellular stresses including infection, trauma, and oxidative damage. However, if uncompensated, the UPR triggers apoptotic cell death. We demonstrate that the three signaling branches of UPR including the PERK, ATF6, and IRE1alpha are rapidly initiated in a mouse model of contusive SCI specifically at the injury epicenter. Immunohistochemical analyses of the various UPR markers revealed that in neurons, the UPR appeared at 6 and 24-h post-SCI. In contrast, in oligodendrocytes and astroglia, UPR persisted at least for up to 3 days post-SCI. The UPR-associated proapoptotic transcriptional regulator CHOP was among the UPR markers upregulated in neurons and oligodendrocytes, but not in astrocytes, of traumatized mouse spinal cords. To directly analyze its role in SCI, WT and CHOP null mice received a moderate T9 contusive injury. Deletion of CHOP led to an overall attenuation of the UPR after contusive SCI. Furthermore, analyses of hindlimb locomotion demonstrated a significant functional recovery that correlated with an increase in white-matter sparing, transcript levels of myelin basic protein, and Claudin 11 and decreased oligodendrocyte apoptosis in CHOP null mice in contrast to WT animals. Thus, our study provides evidence that the UPR contributes to oligodendrocyte loss after traumatic SCI. PMID- 21638342 TI - Metamaterials on paper as a sensing platform. PMID- 21638343 TI - Superhydrophobic-superoleophilic polythiophene films with tunable wetting and electrochromism. PMID- 21638344 TI - Nano gives the answer: breaking the bottleneck of internal concentration polarization with a nanofiber composite forward osmosis membrane for a high water production rate. PMID- 21638345 TI - Remote controlled multishape polymer nanocomposites with selective radiofrequency actuations. PMID- 21638346 TI - Deterministic nanotexturing by directional photofluidization lithography. PMID- 21638347 TI - Solution-processed nanocrystal quantum dot tandem solar cells. PMID- 21638348 TI - Visible-color-tunable light-emitting diodes. PMID- 21638349 TI - Reversible sodium ion insertion in single crystalline manganese oxide nanowires with long cycle life. PMID- 21638350 TI - Facile synthetic route for thickness and composition tunable hollow metal oxide spheres from silica-templated coordination polymers. PMID- 21638353 TI - Identification and quantification of glutathione adducts of clozapine using ultra high-performance liquid chromatography with orthogonal acceleration time-of flight mass spectrometry and inductively coupled plasma mass spectrometry. AB - The application of sulphur-specific detection via ultra-performance liquid chromatography coupled to inductively coupled plasma mass spectrometry (UPLC/ICPMS) to detect and quantify the glutathione (GSH)-adducts produced via the in vitro formation of reactive metabolites is demonstrated. The adducts were formed in human liver microsomes supplemented with unlabelled GSH for clozapine. The calculation of adduct concentration was performed via comparison of the peak areas to calibration curves constructed from omeprazole, a sulphur-containing compound over the range of 0.156 to 15.62 MUM of sulphur with a detection limit of 1.02 ng of sulphur on-column. Identification of the adducts was performed using conventional UPLC/time-of-flight (TOF)-MS with the calculation of clozapine intrinsic clearance carried out by high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS). The use of ICPMS in this way appears to offer a novel, rapid and sensitive means of determining the quantity of GSH conjugates with the combined adducts producing 0.9 MUM of reactive metabolite out of a total of 3.5 MUM of metabolites. The GSH adduct therefore represents 26% of this total produced as a result of the metabolism of drug to reactive species. PMID- 21638354 TI - Effects of formalin preservation on stable carbon and nitrogen isotope signatures in Calanoid copepods: implications for the use of Continuous Plankton Recorder Survey samples in stable isotope analyses. AB - Preserved and archived organic material offers huge potential for the conduct of retrospective and long-term historical ecosystem reconstructions using stable isotope analyses, but because of isotopic exchange with preservatives the obtained values require validation. The Continuous Plankton Recorder (CPR) Survey is the most extensive long-term monitoring program for plankton communities worldwide and has utilised ships of opportunity to collect samples since 1931. To keep the samples intact for subsequent analysis, they are collected and preserved in formalin; however, previous studies have found that this may alter stable carbon and nitrogen isotope ratios in zooplankton. A maximum ~0.90/00 increase of delta(15) N and a time dependent maximum ~1.00/00 decrease of delta(13) C were observed when the copepod, Calanus helgolandicus, was experimentally exposed to two formalin preservatives for 12 months. Applying specific correction factors to delta(15) N and delta(13) C values for similarly preserved Calanoid species collected by the CPR Survey within 12 months of analysis may be appropriate to enable their use in stable isotope studies. The isotope values of samples stored frozen did not differ significantly from those of controls. Although the impact of formalin preservation was relatively small in this and other studies of marine zooplankton, changes in isotope signatures are not consistent across taxa, especially for delta(15) N, indicating that species-specific studies may be required. PMID- 21638355 TI - Characterization of carbonyl by-products during Uniblu-A ozonation by liquid chromatography/hybrid quadrupole time-of-flight/mass spectrometry. AB - The structural elucidation of carbonyl-containing by-products arising from Uniblu OH ozonation has been investigated by liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) employing a quadrupole time-of flight mass spectrometer. The by-products were derivatized with 2,4 dinitrophenylhydrazine, allowing the formation of [M-H](-) ions of the derivatives in the electrospray source. Exact mass measurements of both the [M H](-) ions and their product ions allowed the elemental formulae and related structures of ten by-products to be determined confidently. The main degradation pathway were decarboxylation followed by further oxidation. It is noteworthy that the experimental procedure employed allowed the identification of both nitrogen- and sulphur-containing carbonyl by-products during Uniblu-OH ozonation. This result is of environmental relevance for monitoring the balance of organic nitrogen and sulphur during the ozonation of organic pollutants. These atoms, in fact, do not undergo complete mineralization. PMID- 21638356 TI - Investigating the provenance of un-dyed spun cotton fibre using multi-isotope profiles and chemometric analysis. AB - The analysis of un-dyed spun cotton fibres can be challenging within a forensic science context where discrimination of one fibre from another is of importance. Conventional microscopic and chemical analysis of these fibres is generally unsuccessful because of their similar morphology. In this work we have explored the potential of isotope ratio mass spectrometry (IRMS) as a tool for spun cotton fibre analysis in an attempt to reveal any discriminatory information available. Seven different batches of un-dyed spun cotton fibre from four different countries were analysed. A combination of the hydrogen and oxygen isotopic data facilitated the correct association of the samples, demonstrating, for the first time, the applicability of IRMS to fibre analysis in this way. PMID- 21638357 TI - Identification of unsaturated N-acylhomoserine lactones in bacterial isolates of Rhodobacter sphaeroides by liquid chromatography coupled to electrospray ionization-hybrid linear ion trap-Fourier transform ion cyclotron resonance mass spectrometry. AB - The identification of two unsaturated N-acylhomoserine lactones (AHLs) produced by Rhodobacter sphaeroides bacteria, based on liquid chromatography (LC) coupled to a hybrid quadrupole linear ion trap (LTQ)-Fourier transform ion cyclotron resonance (FTICR) mass spectrometer upon electrospray ionization (ESI), is presented. Besides the confirmation of the signaling molecule already described in the literature, i.e. (Z)-N-tetradec-7-enoyl-homoserine lactone (C(14:1)-HSL), we have discovered the occurrence, at low, yet significant levels, of another monounsaturated compound, C(12:1) -HSL, which may extend the number of small diffusible chemical signals known for R. sphaeroides. Both unsaturated AHLs were identified by high-resolution FTICR mass spectrometry in extracts of bacterial culture media and the occurrence of a C=C bond was assessed upon their conversion into bromohydrins. Collision-induced dissociation (CID) spectra were then collected on the LTQ mass analyzer. A careful comparison of tandem MS spectra of monounsaturated (i.e., C(12:1)-HSL and C(14:1)-HSL) and saturated AHLs (i.e. C(12)-HSL and C(14)-HSL) led to the emphasis of two series of product ions, exhibiting 14 Da spaced m/z ratios. Both series were referred to progressive fragmentations at the aliphatic end of the AHL acyl chains, followed by neutral losses of terminal alkenes (i.e. CH(2)=CH(CH(2))(n)H). In particular, the series located at the higher end of the explored m/z range (>200 Da), observed only for monounsaturated species, enabled the location of the C=C bond between carbons 7 and 8 of the acyl chain. PMID- 21638358 TI - Assessment of effects of the rising atmospheric nitrogen deposition on nitrogen uptake and long-term water-use efficiency of plants using nitrogen and carbon stable isotopes. AB - This study assesses the effects of the atmospheric nitrogen (N) deposition on the N uptake and the long-term water-use efficiency of two C(3) plants (Agropyron cristatum and Leymus chinensis) and two C(4) plants (Amaranthus retroflexus and Setaria viridis) using N and C stable isotopes. In addition, this study explores the potential correlation between leaf N isotope (delta(15)N) values and leaf C isotope (delta(13)C) values. This experiment shows that the atmospheric N deposition has significant effects on the N uptake, delta(15)N and leaf N content (N(m)) of C(3) plants. As the atmospheric N deposition rises, the proportion and the amount of N absorbed from the simulated atmospheric deposition become higher, and the delta(15)N and N(m) of the two C(3) plants both also increase, suggesting that the rising atmospheric N deposition is beneficial for C(3) plants. However, C(4) plants display different patterns in their N uptake and in their variations of delta(15)N and N(m) from those of C(3) plants. C(4) plants absorb less N from the atmospheric deposition, and the leaf N(m) does not change with the elevated atmospheric N deposition. Photosynthetic pathways may account for the differences between C(3) and C(4) plants. This study also shows that atmospheric N deposition does not play a role in determining the delta(13)C and in the long-term water-use efficiency of C(3) and C(4) plants, suggesting that the long-term water-use pattern of the plants does not change with the atmospheric N input. In addition, this study does not observe any relationship between leaf delta(15)N and leaf delta(13)C in both C(3) and C(4) plants. PMID- 21638359 TI - Infrared multiple-photon dissociation spectroscopy of group II metal complexes with salicylate. AB - Ion trap tandem mass spectrometry with collision-induced dissociation, and the combination of infrared multiple-photon dissociation (IRMPD) spectroscopy and density functional theory (DFT) calculations, were used to characterize singly charged, 1:1 complexes of Ca(2+), Sr(2+) and Ba(2+) with salicylate. For each metal-salicylate complex, the CID pathways are: (a) elimination of CO(2) and (b) formation of [MOH](+) where M = Ca(2+), Sr(2+) or Ba(2+). DFT calculations predict three minima for the cation-salicylate complexes which differ in the mode of metal binding. In the first, the metal ion is coordinated by O atoms of the (neutral) phenol and carboxylate groups of salicylate. In the second, the cation is coordinated by phenoxide and (neutral) carboxylic acid groups. The third mode involves coordination by the carboxylate group alone. The infrared spectrum for the metal-salicylate complexes contains a number of absorptions between 1000 and 1650 cm(-1), and the best correlation between theoretical and experimental spectra is found for the structure that features coordination of the metal ion by phenoxide and the carbonyl O of the carboxylic acid group, consistent with the calculated energies for the respective species. PMID- 21638360 TI - Metabolomic investigation of cholestasis in a rat model using ultra-performance liquid chromatography/tandem mass spectrometry. AB - Metabolomics follows the changes in concentrations of endogenous metabolites, which may reflect various disease states as well as systemic responses to environmental, therapeutic, or genetic interventions. In this study, we applied metabolomic approaches to monitor dynamic changes in plasma and urine metabolites, and compared these metabolite profiles in Eisai hyperbilirubinemic rats (EHBR, an animal model of cholestasis) with those in the parent strain of EHBR - Sprague-Dawley (SD) rats - in order to characterize cholestasis pathophysiologically. Ultra-performance liquid chromatography/tandem mass spectrometry-based analytical methods were used to assay metabolite levels. More than 250 metabolites were detected in both plasma and urine, and metabolite profiles of EHBR differed from those of SD rats. The levels of antioxidative and cytoprotective metabolites, taurine and hypotaurine, were markedly increased in urine of EHBR. The levels of many bile acids were also elevated in plasma and urine of EHBR, but the extent of elevation depended on the particular bile acid. The levels of cytoprotective ursodeoxycholic acid and its conjugates were markedly elevated, while that of cytotoxic chenodeoxycholic acid remained unchanged, suggesting the balance of bile acids had shifted resulting in decreased toxicity. In EHBR, reduced biliary excretion leads to increased systemic exposure to harmful compounds including some endogenous metabolites. Our metabolomic data suggest that mechanisms exist in EHBR that compensate for cholestasis-related damage. PMID- 21638361 TI - The effects of preservation methods, dyes and acidification on the isotopic values (delta15N and delta13C) of two zooplankton species from the KwaZulu-Natal Bight, South Africa. AB - Stable isotope measurements are an important tool for ecosystem trophic linkage studies. Ideally, fresh samples should be used for isotopic analysis, but in many cases organisms must be preserved and analysed later. In some cases dyes must be used to help distinguish organisms from detritus. Since preservatives and dyes are carbon-based, their addition could influence isotopic readings. This study aims to improve understanding of the effects of sample storage method, dye addition and acidification on the delta(15)N and delta(13)C values of zooplankton (Euphasia frigida and Undinula vulgaris). Zooplankton was collected and preserved by freezing, or by the addition of 5% formalin, 70% ethanol, or 5% formalin with added Phloxine B or Rose Bengal, and stored for 1 month before processing. Samples in 5% formalin and 70% ethanol were also kept and processed after 3 and 9 months to study changes over time. Formalin caused the largest enrichment for delta(13)C and a slight enrichment for delta(15)N, while ethanol produced a slight depletion for delta(13)C, and different effects on delta(15)N depending on the species. In formalin, dyes depleted the delta(13)C values, but had variable effects on delta(15)N, relative to formalin alone. Acidification had no significant effect on delta(15)N or delta(13)C for either species. Long-term storage showed that the effects of the preservatives were species-dependent. Although the effects on delta(15)N varied, a relative enrichment in (13)C of samples occurred with time. This can have important consequences for the understanding of the organic flow within a food web and for trophic studies. . PMID- 21638362 TI - Efficient enrichment and identification of phosphopeptides by cerium oxide using on-plate matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis. AB - An efficient and simple method for enrichment and identification of phosphopeptides by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) using cerium oxide is presented. After pretreatment of tryptic digests of phosphoproteins with CeO(2), nonphosphopeptides are discarded and phosphopeptides are enriched. By applying the separated CeO(2) on a target plate and analysis using MALDI-TOF MS, peaks of phosphopeptides and their correspondingly series of dephosphorylated peptides are observed in the mass spectra. Thus, the phosphopeptides are very easy to identify with the mass difference, which are all 80 Da between adjacent peaks in the same series, and clear background in the spectra owing to elimination of signal suppression from large amounts of nonphosphopeptides. Furthermore, the phosphopeptides can be dephosphorylated completely after a further NH(4)OH elution. Tryptic digest products from several standard proteins are pretreated using CeO(2) to demonstrate the efficiency of this method. Phosphopeptides from a very small quantity of human serum are enriched and analyzed, and proteins also identified by searching against a database using Mascot on MALDI-TOF/TOF fragments, which indicates that this method may be employed in complex samples for further application. PMID- 21638363 TI - Development and in-house validation of a robust and sensitive solid-phase extraction liquid chromatography/tandem mass spectrometry method for the quantitative determination of aflatoxins B1, B2, G1, G2, ochratoxin A, deoxynivalenol, zearalenone, T-2 and HT-2 toxins in cereal-based foods. AB - A sensitive and robust liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was developed for the simultaneous determination of aflatoxins (B(1), B(2), G(1), G(2)), ochratoxin A, deoxynivalenol, zearalenone, T-2 and HT-2 toxins in cereal-based foods. Samples were extracted with a mixture of acetonitrile/water (84:16, v/v) and cleaned up through a polymeric solid-phase extraction column. Detection and quantification of the nine mycotoxins were performed by reversed-phase liquid chromatography coupled with electrospray ionization triple quadrupole mass spectrometry (LC/ESI-MS/MS), using fully (13)C isotope-labelled mycotoxins as internal standards. The method was validated in house for five different cereal processed products, namely barley, oat and durum wheat flours, rye- and wheat-based crisp bread. Recoveries and repeatability of the whole analytical procedure were evaluated at contamination levels encompassing the EU maximum permitted levels for each tested mycotoxin. Recoveries ranged from 89 to 108% for deoxynivalenol, from 73 to 114% for aflatoxins, from 85 to 114% for T-2 and HT-2 toxins, from 64 to 97% for zearalenone, from 74 to 102% for ochratoxin A. Relative standard deviations were less than 16% for all tested mycotoxins and matrices. Limits of detection (signal to-noise ratio 3:1) ranged from 0.1 to 59.2 ug/kg. The trueness of the results obtained by the proposed method was demonstrated by analysis of reference materials for aflatoxins, deoxynivalenol, zearalenone. The use of inexpensive clean-up cartridges and the increasing availability of less expensive LC/MS/MS instrumentation strengthen the potential of the proposed method for its effective application for reliable routine analysis to assess compliance of tested cereal products with current regulation. PMID- 21638364 TI - Evaluation of combined matrix-assisted laser desorption/ionization time-of-flight and matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry experiments for peptide mass fingerprinting analysis. AB - Peptide Mass Fingerprinting (PMF) is still of significant interest in proteomics because it allows a large number of complex samples to be rapidly screened and characterized. The main part of post-translational modifications is generally preserved. In some specific cases, PMF suffers from ambiguous or unsuccessful identification. In order to improve its reliability, a combined approach using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) and matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICRMS) was evaluated. The study was carried out on bovine serum albumin (BSA) digest. The influence of several important parameters (the matrix, the sample preparation method, the amount of the analyte) on the MOWSE score and the protein sequence coverage were evaluated to allow the identification of specific effects. A careful investigation of the sequence coverage obtained by each kind of experiment ensured the detection of specific peptides for each experimental condition. Results highlighted that DHB-FTICRMS and DHB- or CHCA-TOFMS are the most suited combinations of experimental conditions to achieve PMF analysis. The association (convolution) of the data obtained by each of these techniques ensured a significant increase in the MOWSE score and the protein sequence coverage. PMID- 21638365 TI - Gas chromatographic mass spectrometric detection of dihydroxy fatty acids preserved in the 'bound' phase of organic residues of archaeological pottery vessels. AB - A methodology is presented for the determination of dihydroxy fatty acids preserved in the 'bound' phase of organic residues preserved in archaeological potsherds. The method comprises saponification, esterification, silica gel column chromatographic fractionation, and analysis by gas chromatography/mass spectrometry. The electron ionisation mass spectra of the trimethylsilyl ether methyl ester derivatives are characterised by fragment ions arising from cleavage of the bond between the two vicinal trimethylsiloxy groups. Other significant fragment ions are [M-15](+.), [M-31](+.), m/z 147 and ions characteristic of vicinal disubstituted (trimethylsiloxy) TMSO- groups (Delta(7,8), Delta(9,10), Delta(11,12) and Delta(13,14): m/z 304, 332, 360 and 388, respectively). The dihydroxy fatty acids identified in archaeological extracts exhibited carbon numbers ranging from C(16) to C(22) and concentrations varying from 0.05 to 14.05 ug g(-1) . The wide range of dihydroxy fatty acids observed indicates that this approach may be applied confidently in screening archaeological potsherds for the degradation products of monounsaturated fatty acids derived from commodities processed in archaeological pottery vessels. PMID- 21638366 TI - Preliminary fingerprinting of Athabasca oil sands polar organics in environmental samples using electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry. AB - There is a growing need to develop analytical methods that can distinguish compounds found within industrially derived oil sands process water (OSPW) from those derived from natural weathering of oil sands deposits. This is a difficult challenge as possible leakage beyond tailings pond containments will probably be in the form of mixtures of water-soluble organics that may be similar to those leaching naturally into aquatic environments. We have evaluated the potential of negative ion electrospray ionization high-resolution Fourier transform ion cyclotron resonance mass spectrometry (FTICRMS) for comparing oil sands polar organics from tailing ponds, interceptor wells, groundwater, river and lake surface waters. Principal component analysis was performed for all species observed. which included the O(2) class (often assumed to be monocarbxoylic naphthenic acids) along with a wide range of other species including humic substances in the river and lake samples: O(n) where n=1-16; NO(n) and N(2)O(n) where n=1-13; and O(n)S and O(n)S(2) where n=1-10 and 1-8, respectively. A broad range of species was investigated because classical naphthenic acids can be a small fraction of the 'organics' detected in the polar fraction of OSPW, river water and groundwater. Aquatic toxicity and environmental chemistry are attributed to the total organics (not only the classical naphthenic acids). The distributions of the oil sands polar organics, particularly the sulfur-containing species, O(n)S and O(n)S(2), may have potential for distinguishing sources of OSPW. The ratios of species containing O(n) along with nitrogen-containing species: NO(n), and N(2)O(n), were useful for differentiating organic components derived from OSPW from those found in river and lake waters. Further application of the FTICRMS technique for a diverse range of OSPW of varying ages and composition, as well as the surrounding groundwater wells, may be critical in assessing whether leakage from industrial sources to natural waters is occurring. PMID- 21638367 TI - Carbon and oxygen isotope analysis of small carbonate samples (20 to 100 ug) with a GasBench II preparation device. PMID- 21638369 TI - Macromol. Rapid commun. 14/2009. PMID- 21638368 TI - Comparison of ex-vivo high-resolution episcopic microscopy with in-vivo four dimensional high-resolution transvaginal sonography of the first-trimester fetal heart. AB - OBJECTIVE: To compare the capability of three-dimensional (3D) reconstructed images produced by high-resolution episcopic microscopy (HREM) with that of in vivo four-dimensional high-resolution transvaginal sonography (4D-HRTVS) to discern morphological features of the first-trimester human fetal heart. METHODS: This was a prospective study of fetal hearts between 9 and 14 weeks' gestation. For ex-vivo 3D analysis, 30 human fetal hearts (at 9 + 0 to 14 + 6 weeks) were retrieved from surgical terminations of pregnancy. The specimens were embedded in resin and episcopic ('block-face') imaging was used to obtain a digital volume dataset (HREM) using 3-micron slicing. 4D-HRTVS was performed in 28 separate pregnancies at 10 + 2 to 14 + 0 weeks using a Voluson E8 ultrasound machine with volumetric transvaginal RIC 6-12-MHz transducers. Heart volumes obtained by both methods were compared to assess their ability to demonstrate first-trimester cardiac morphology. Comparisons were made in the transverse and sagittal planes, and using volume rendering. RESULTS: All hearts were structurally normal, although abdominal situs was not examined in the isolated hearts that underwent HREM. 4D-HRTVS demonstrated each of the complete five transverse cardiac views in 32-86% of cases. HREM showed four features unique to the first-trimester human heart: prominent atrial appendages, spiral ventricular arrangement, prominent coronary arteries and thickened arterial walls. 4D-HRTVS could demonstrate the first two, but ultrasound resolution was too poor to quantify wall thickness and demonstrate coronary arteries in the 3-5-mm diameter heart. CONCLUSIONS: 4D-HRTVS showed limited morphological features of the first-trimester fetal heart compared with HREM. HREM provides a gold standard of ex-vivo imaging against which developments in ultrasound resolution could be compared. PMID- 21638370 TI - Self-organized materials for optoelectronic applications. PMID- 21638371 TI - Templated organic and hybrid materials for optoelectronic applications. AB - The review highlights different approaches to template organic materials as well as hybrid materials that find or are expected to find application in optoelectronic devices. The first templating approach focuses on the use of preformed nanoporous membranes as templates for organic materials and polymeric materials. Such nanoporous templates can be track-etched membranes, anodic aluminum oxide membranes and other variants thereof, or block copolymer templates. Further, opals have been described as templates. In the second part, we have summarized developments that take advantage of self-assembly processes to pattern hybrid materials. Examples are sol-gel templating techniques using amphiphiles, evaporation-induced self-assembly, lyotropic templating as well as templating from block copolymers. Both routes are very promising templating approaches for optoelectronic materials and represent complementary rather than competing techniques. PMID- 21638372 TI - Electrical modes in scanning probe microscopy. AB - Scanning probe microscopy methods allow the investigation of a variety of sample surface properties on a nanometer scale, even down to single molecules. As molecular electronics advance, the characterization of electrical properties becomes more and more important. In both research and industry, films made from composite materials and lithographically structured elements have already reached structure sizes down to a few nanometers. Here, we review the major scanning probe microscopy modes that are used for electrical characterization of thin films, that is, scanning conductive force microscopy, Kelvin probe force microscopy and scanning electric field microscopy. To demonstrate the possibilities and capabilities of these modes, reference samples were fabricated by means of focused ion beam deposition and analyzed using the described methods. Furthermore, two upcoming modes are presented that are based on: i) local current measurements while the SPM-cantilever is excited into torsional vibrations, and, ii) changes in a backscattered microwave that was coupled into a scanning probe microscopy-cantilever. The scanning-probe-based electrical modes are applicable for studies of functional layers used in soft matter electronic devices under realistic environmental conditions. PMID- 21638373 TI - Liquid crystalline ordering and charge transport in semiconducting materials. AB - Organic semiconducting materials offer the advantage of solution processability into flexible films. In most cases, their drawback is based on their low charge carrier mobility, which is directly related to the packing of the molecules both on local (amorphous versus crystalline) and on macroscopic (grain boundaries) length scales. Liquid crystalline ordering offers the possibility of circumventing this problem. An advanced concept comprises: i) the application of materials with different liquid crystalline phases, ii) the orientation of a low viscosity high temperature phase, and, iii) the transfer of the macroscopic orientation during cooling to a highly ordered (at best, crystalline-like) phase at room temperature. At the same time, the desired orientation for the application (OLED or field-effect transistor) can be obtained. This review presents the use of molecules with discotic, calamitic and sanidic phases and discusses the sensitivity of the phases with regard to defects depending on the dimensionality of the ordered structure (columns: 1D, smectic layers and sanidic phases: 2D). It presents ways to systematically improve charge carrier mobility by proper variation of the electronic and steric (packing) structure of the constituting molecules and to reach charge carrier mobilities that are close to and comparable to amorphous silicon, with values of 0.1 to 0.7 cm(2) . V(-1) . s(-1) . In this context, the significance of cross-linking to stabilize the orientation and liquid crystalline behavior of inorganic/organic hybrids is also discussed. PMID- 21638374 TI - Excitation energy transfer in organic materials: from fundamentals to optoelectronic devices. AB - In this review, we discuss investigations of electronic excitation energy transfer in conjugated organic materials at the bulk and single molecule level and applications of energy transfer in fluorescent and phosphorescent organic light emitting devices. A brief overview of common descriptions of energy transfer mechanisms is given followed by a discussion of some basic photophysics of conjugated materials including the generation of excited states and their subsequent decay through various channels. In particular, various examples of bimolecular excited state annihilation processes are presented. Energy transfer studies at the single molecule level provide a new tool to study electronic couplings in simple donor/acceptor dyads and conjugated polymers. Finally, energy transfer in organic electronic devices is discussed with particular emphasis on triplet emitter doped OLEDs and blends for white light emission. PMID- 21638375 TI - Tunable Phosphorescent Emission through Energy Transfer within Multilayer Thin Films Based on a Carbazole-Based Host and Ir(III)-Complex Guest System. AB - A new method to tune both phosphorescence emission intensity and spectroscopic colors based on the alternatively structured host/guest multilayer thin films prepared by the spin-assisted LbL deposition is presented. Their emission characteristics are demonstrated with pairs of positively charged Ir-PEI complexes as guests and negatively charged CBZ-PAA as hosts. The phosphorescent emission of Ir-PEI complexes is enhanced by the energy transfer from the host to the guest and, additionally, this energy transfer can be finely tuned by the insertion of spacer layers between the phosphorescent donor and acceptor layers to vary the emission intensity as well as to render different emission colors. PMID- 21638376 TI - Surface coatings based on polysilsesquioxanes: solution-processible smooth hole injection layers for optoelectronic applications. AB - Optoelectronic devices usually consist of a transparent conductive oxide (TCO) as one electrode. Interfacial engineering between the TCO electrode and the overlying organic layers is an important method for tuning device performance. We introduce poly(methylsilsesquioxane)-poly(N,N-di-4-methylphenylamino styrene) (PMSSQ-PTPA) as a potential hole-injection layer forming material. Spin-coating and thermally induced crosslinking resulted in an effective planarization of the anode interface. HOMO level (-5.6 eV) and hole mobility (1 * 10(-6) cm(2) . Vs( 1) ) of the film on ITO substrates were measured by cyclovoltammetry and time-of flight measurement demonstrating the hole injection capability of the layer. Adhesion and stability for further multilayer built-up could be demonstrated. Contact angle measurements and tape tests after several solvent treatments proved the outstanding film stability. PMID- 21638377 TI - Semiconductor dendritic-linear block copolymers by nitroxide mediated radical polymerization. AB - The first synthesis of a semiconductor hybrid diblock copolymer comprised of p type dendritic and n-type linear blocks by nitroxide mediated radical polymerization (NMRP) is reported. A triphenylamine (TPA) bearing second generation polyether dendron [G2]-OH has been functionalized with an alkoxyamine and, subsequently, perylene bisimide acrylate (PerAcr) was polymerized to obtain a hybrid block copolymer, [G2]-b-PPerAcr. The hybrid block copolymer structure is supported by (1) H NMR and size exclusion chromatography. Furthermore, the novel materials were studied by UV-vis absorption spectrometry, photoluminescence, cyclic voltammetry, differential scanning calorimetry and thermogravimetry analysis. PMID- 21638378 TI - Polymerizable Well-Defined Oligo(thiophene amide)s and their ROMP Block Copolymers. AB - We report the synthesis of conjugated thiophene amide oligomers that constitute a new class of chromophores with potential for optoelectronic applications. The synthesis of defined norbornene-substituted oligothiophene amides using conventional coupling chemistry is described. Their electronic properties depend on the degree of oligomerization as UV/Vis and fluorescence spectroscopy demonstrate. A significant red shift in the spectra upon an increase in the oligomer length evidences conjugation of the thiophene rings via the amide linkages. ROMP of the norbornene-substituted oligomers gives homopolymers and block-copolymers with a solubilizing second block. The amphiphilic character of the block copolymers is used to study micellization and bulk self-organization. PMID- 21638379 TI - Amorphous Carbazole-based (Co)polymers for OFET Application. AB - In this contribution, we present the synthesis and characterization of poly[9-(4 alkylphenyl)carbazole-3,6-diyl] and poly[1,3-bis(3'-alkylcarbazole-9'-yl)-5 alkylphenylene-6',6"-diyl]. The amorphous solid state structure, reasonably good field effect mobility and on/off ratios of these novel carbazole-based polymers make them promising candidates as hole conducting materials for printed organic field-effect transistors (OFETs). PMID- 21638380 TI - Relationship of photophysical properties and the device performance of novel hybrid small-molecular/polymeric solar cells. AB - We investigate solar cells comprised of a vinazene derivative (HV-BT) as the electron acceptor and the well-known polymer poly(3-hexylthiophene) as the electron donor. In the as-prepared blend, most of the excited state species, including the excimers on HV-BT, are quenched at the heterojunction. Although the photophysical properties of the blends change upon annealing, the blend solar cells largely remain uninfluenced by such treatments. A significant improvement is, however, observed when inducing phase separation at a longer length scale, for example, in solution-processed bilayer devices. Hereby, both the fill factor (FF) and the open circuit voltage are considerably increased, pointing to the importance of the heterojunction topology and the layer composition at the charge extracting contacts. An optimized device exhibits a power conversion efficiency of close to 1%. PMID- 21638381 TI - Real Time Investigation of the Interface between a P3HT:PCBM Layer and an Al Electrode during Thermal Annealing. AB - Real time variation of the interfacial structure between an Al electrode and a poly(3-hexylthiophene) (P3HT):fullerene (PCBM) thin film during thermal annealing has been investigated using synchrotron X-rays. It is found that Al atoms diffuse into the organic layer to form a thin interlayer between the Al electrode and the organic layer even during the deposition of an Al layer. The interlayer thickness and the mass density of the interlayer increases if annealed above 120 degrees C. The interlayer thickness depends on the annealing processes and the inter diffusion is accelerated by a fast annealing process. The Al diffusion reduces the preferred alignment of the (100) direction of the P3HT crystals from the surface normal direction and randomizes their orientation. The Al diffusion also helps to reduce the contact resistance in the P3HT:PCBM-based solar cells. PMID- 21638382 TI - Hetero-telechelic dye-labeled polymer for nanoparticle decoration. AB - The synthesis of poly(methyl methacrylate) (PMMA) exhibiting one fluorescent dye (Texas Red) and one methyl disulfide end group is described. It is shown that the latter end group enabled the exchange of both oleic amine on gold nanoparticles (AuNP) and of oleic acid on CdSe/ZnS quantum dots (QD), allowing for a phase transfer of both types of nanoparticles (NP) from hexane into dimethylformamide due to the solubility provided by the PMMA chains. For AuNP, a fluorescence quenching of the dye was found due to fluorescence resonance energy transfer (FRET) from the dye to the AuNP, while QDs caused a fluorescence enhancement by FRET from the QD to the attached dyes. Due to the hetero-telechelic geometry of the polymer, the separation between NP and dye is governed by the end-to-end distance of the polymer. PMID- 21638383 TI - Conjugated Triphenylene Polymers for Blue OLED Devices. AB - Three polytriphenylene derivatives with phenyl and alkyl groups as side chains have been tested as blue light emitters. The best performance in terms of turn-on voltage and luminance efficiencies was obtained for poly(2-heptyl-3-(4 octylphenyl)-1,4-diphenyl-6,11-triphenylenyl-1,4-benzene) (1), which was additionally blended with an electron-transporting and a hole-injection material in the device, and had an onset of 4.6 V and reached 0.73 cd . A(-1) . PMID- 21638384 TI - Macromol. Rapid commun. 15/2009. PMID- 21638385 TI - The Gilch Synthesis of Poly(p-phenylene vinylenes): Mechanistic Knowledge in the Service of Advanced Materials. AB - A consistent picture is presented of the mechanistic details and intermediates of the Gilch polymerization leading to poly(p-phenylene vinylenes) (PPVs). In-situ generated p-quinodimethanes are shown to be the real monomers, and spontaneous formation of the initiating radicals is effected by dimerization of some of these monomers to dimer diradicals, the latter also being the reason why significant amounts of [2.2]paracyclophanes are formed as side-products. Chain propagation predominantly proceeds by radical chain growth, occasionally interrupted by polyrecombination events between the growing alpha,omega-macro-diradicals. Based on this knowledge, oxygen is identified as a very efficient molar-mass regulating agent, and the temporary gelation of the reaction mixtures is interpreted to be the consequence of a very high entanglement of the polymers immediately after their formation. Last but not least, it is rationalized why the usually considered constitutional defects in Gilch PPVs might not be the only and most relevant ones with respect to the efficiency and durability of the organic light emitting devices produced thereof, and why cis-configurated halide-bearing vinylene moieties should be perceived as being among the most critical candidates. These considerations result in the recommendation of straightforward measures that should lead to clearly improved PPVs. PMID- 21638386 TI - Design of soluble hyperbranched polythiophenes with tailor-made optoelectronic properties. AB - Hyperbranched polythiophenes were prepared via a simple one-pot synthesis approach based on oxidative coupling of branched conjugated monomers. Only small variations in the building unit and architecture lead to large differences of absorption and photoluminescence properties. Interestingly, soluble hyperbranched polythiophenes with relatively small molecular weights show enhanced absorption at low and high wavelengths compared to linear analogues, such as poly(3-hexyl thiophenes) with high molecular weights. With this versatile approach we present a method to design tailor made, functional materials with potential applications in optoelectronics. PMID- 21638387 TI - 'Click' functionalization of cryogels conveniently verified and quantified using high-resolution MAS NMR spectroscopy. AB - Chemical modification reactions of alkyne containing polyHEMA-based macroporous network structures (cryogels) by Cu(I) catalyzed azide-alkyne 'click' cycloaddition reactions and their monitoring and quantification with high resolution magic angle spinning (hr-MAS) NMR spectroscopy are reported. Complete conversion is obtained when benzylazide is reacted with the grafted alkyne function, but only partial conversion is observed when using azide-modified poly(ethylene glycol) (PEG-N(3) ). Subsequent addition of benzylazide consumes all remaining alkyne groups. All chemical modifications are easily monitored at each stage using hr-MAS NMR spectroscopy. The alkyne functionality and the resulting triazole ring provide well resolved (1) H resonances to monitor and quantify the progress of such 'click' reactions in general. PMID- 21638388 TI - Control of Induced Chirality in Optically Active Poly(N-propargylcarbamate) Films by Solvent Vapor. AB - Chiral polyethyne derivatives with lyotropic liquid-crystalline properties are found to be able to self-assemble, forming two chiral organizations with opposite handedness in solid thin films by selection of the casting solvent and its concentration. After the film preparation, chiral organization could also be induced by simple exposure to an appropriate organic solvent's vapor for several minutes without thermal treatment. Furthermore, irreversible inversion of the handedness of the chiral organization in the film could be achieved by exposure to solvent vapor. PMID- 21638389 TI - Synthesis of a fluorescent polymer bearing covalently linked thienylene moieties and rhodamine for efficient sensing. AB - A novel conjugated polymer (RB-PPETE) of poly[p-(phenylene ethynylene)-alt (thienylene ethynylene)] (PPETE) bearing covalently linked thienylene rings and Rhodamine B units has been synthesized and successfully used to detect metal ions. The Rhodamine B exists as a lactone, which is colorless and non fluorescent. Hg(2+) ions can induce the Rhodamine group to form a ring-opened state. The fluorescence resonance energy transfer (FRET) was demonstrated in the polymer, and in the presence of Hg(2+) ions the excitation energy along the backbone of the conjugated polymer is transferred to the energy acceptor (Rhodamine B), which leads to a visual color change of the solution from slight yellow to orange. Meanwhile, this new system shows outstanding Hg(2+) -selective FRET off-on type fluoroionophoric properties among the representative metal ions in tetrahydrofuran. PMID- 21638390 TI - Ultrahigh-Density Carbon Nanoring Arrays on Silicon Wafer through Templated Solution Deposition Method. AB - Ultrahigh-density carbon nanoring arrays on a silicon wafer are achieved by a novel templated solution deposition method. Initially the silica nanodot arrays obtained from a nanoporous thin film are used as a template to direct the surface dewetting of a phenolic precursor, while further curing and calcination of the phenolic precursor, followed by etching of the silica arrays, results in large area carbon nanoring arrays with a diameter as small as 25 nm. This study provides a simple and robust chemical route to fabricate complex nanoring arrays with ultrahigh density of about one terabit per square inch. PMID- 21638391 TI - One-step preparation of antimicrobial polyrhodanine nanotubes with silver nanoparticles. AB - A simple synthetic method has been developed for the fabrication of antimicrobial polyrhodanine nanotubes with silver nanoparticles. Rhodanine monomer first forms one-dimensional complexes with silver ions due to coordinative interactions and consecutively reduces the silver ions during chemical-oxidation polymerization. The polymerization procedure is analyzed by transmission electron microscopy and scanning electron microscopy in situ. The synthesized silver nanoparticles/polyrhodanine nanotubes are applied as an antimicrobial agent against Gram-negative bacteria, E. coli and Gram-positive bacteria, S. aureus. The antimicrobial tests demonstrate that the silver/polyrhodanine nanotubes have superior antimicrobial properties to silver nanoparticles and rhodanine monomer. PMID- 21638392 TI - Coordination-Polymeric Nanofibers and their Field-Emission Properties. AB - The large-scale fabrication of nanofibers of coordination polymers (CPs) is reported for the first time from CP gel precursors by the electrospinning method. In the absence of organic polymeric additives, viscous solutions suitable for drawing fibers have been made by diluting the gel precursors, which are obtained by mixing Ni(OAc)(2) and 4-trifluoromethylbenzoic acid with 4,4' bis(pyridyl)ethylene (bpe) or 4,4'-bipyridine (bpy) linear spacer ligands in organic solvents. One-dimensional polymeric structures have been proposed for these gels and nanofibers based on the crystal structures of the compounds isolated. These fibers show some interesting field-emission properties. The results described here may well be extended to other one-dimensional polymers, thus opening the door for processing of the coordination polymers. PMID- 21638393 TI - Macromol. Rapid commun. 16/2009. PMID- 21638394 TI - Simultaneous chain-growth and step-growth polymerization-a new route to cyclic polymers. AB - Reinvestigation of numerous ring-opening polymerizations by means of MALDI-TOF mass spectrometry has evidenced that cyclic polymers were formed as the only reaction products or, at least, in large fractions. This finding is ascribed to the intermediate formation of difunctional chains having active end groups that can react with each other. Due to the low concentration of these difunctional chains cyclization is favored over chain extension according to the Ruggli Ziegler dilution principle. A polymerization mechanism which usually favors the formation of cyclic polymers is the zwitterionic polymerization, but an exception from this rule is known. The following classes of monomers were discussed: alpha amino acid, N-carboxyanhydrides (oxazolidine-2,5-diones), dithiolane-2,4-diones, 5,5-dimethyl-1,3,2-dioxathiolan-4-one-2-oxide, salicylic acid O-carboxyanhydride, L-lactide and D,L-lactide, hexamethyl cyclotrisiloxane, and macrocyclic dithiocarbamates. PMID- 21638395 TI - Bottom-up Assembly of Poly(3-hexylthiophene) on Carbon Nanotubes: 2D Building Blocks for Nanoscale Circuits. AB - Hierarchical poly(3-hexylthiophene)(P3HT)/carbon nanotube (CNT) supramolecular structures were fabricated through a bottom-up CNT induced P3HT crystallization strategy. P3HT nanowires growing perpendicular from CNT surface have uniform width and height. The density and the length of these nanowires can be controlled by tuning the P3HT/CNT mass ratio. The quasi-isothermal crystallization process monitored by in situ UV-Vis spectroscopy indicates that CNTs can greatly enhance the P3HT crystallization, and the P3HT nanowire formation follows first-order kinetics. Such bottom-up strategy provides a general approach to build 2D functional conductive supramolecular structures that will lead to numerous applications in nanoscale electronics. PMID- 21638396 TI - Synthesis of comb tri- and tetrablock copolymers catalyzed by the grubbs first generation catalyst. AB - High molecular weight tri- and tetrablock copolymers were synthesized from the commercially available Grubbs first generation catalyst for the first time. These polymers had degrees of polymerization from 430 to 1 100, molecular weights up to 419 000 g . mol(-1) , and narrow polydispersities. Oxanorbornene monomers were chosen due to their fast rates of polymerization and slow rates of cross metathesis. Polystyrene arms were grown from selected blocks by atom transfer radical polymerization to yield architecturally complex comb tri- and tetrablock copolymers. These polymers self-assembled in the solid state into ordered morphologies that were characterized by scanning electron microscopy. PMID- 21638397 TI - Poly(p-phenylene ethynylene) Incorporating Sterically Enshrouding m-Terphenyl Oxacyclophane Canopies. AB - A sterically encumbered m-terphenyl oxacyclophane substituted with two aryl iodide substituents has been prepared as a versatile monomer for the preparation of pi-conjugated polymers. The monomer has been used to prepare a poly(p phenylene ethynylene) derivative (P1) incorporating oxacyclophane units as canopies that shield one side of the pi-system from inter-chain interactions. The photophysical properties of P1 in dilute solution compare well to those of a poly(p-phenylene ethynylene) derivative (P2) that lacks the canopy. The presence of the steric canopy leads to a diminished inter-chain interaction in the solid state and enhances the kinetic response of P1 to vapors of nitro-organics such as TNT, presumably by increasing the permeability of P1 to these analytes over that of P2. PMID- 21638398 TI - Dry Spinning Based Spinneret Based Tunable Engineered Parameters (STEP) Technique for Controlled and Aligned Deposition of Polymeric Nanofibers. AB - Polymeric nanofibers are finding increasing number of applications and hold the potential to revolutionize diverse fields such as tissue engineering, smart textiles, sensors, and actuators. Aligning and producing high aspect ratio fiber arrays (length/diameter > 2 000) in the sub-micron and nanoscale diameters has been challenging due to fragility of polymeric materials, thus making it difficult to deposit them as one dimensional structures functionally interfaced with other systems. Here, we present a pseudo dry spinning technique which allows precise control on fiber diameters and further allows deposition of fiber arrays in aligned configurations. Control on fiber diameters ranging from 50-500 nm and having lengths of several millimeters is achieved by altering the polymeric solution concentration. In the dilute and semi-dilute unentangled concentration domain droplets or beaded fibers are observed to form. Smooth uniform diameter fibers are observed to form at the onset of semi-dilute entangled concentration regime. For a given molecular weight, the increase in fiber diameter with increasing solution concentration is attributed to both the increase in the entanglement density and the decrease in the radius of gyration of solvated polymer molecules. Using this technique polymeric fiber arrays in single and multiple layers are demonstrated which can be used towards developing strong textiles, biological scaffolds, and sensor networks. PMID- 21638399 TI - Synthesis of Well-Defined Polybenzamide-block-Polystyrene by Combination of Chain Growth Condensation Polymerization and RAFT Polymerization. AB - Well-defined diblock copolymers composed of poly(N-octylbenzamide) and polystyrene were synthesized by reversible addition-fragmentation chain transfer (RAFT) polymerization of styrene with a polyamide chain transfer agent (CTA) prepared via chain-growth condensation polymerization. Synthesis of a dithioester type macro-CTA possessing the polyamide segment as an activating group was unsatisfactory due to side reactions and incomplete introduction of the benzyl dithiocarbonyl unit. On the other hand, a dithiobenzoate-CTA containing poly(N octylbenzamide) as a radical leaving group was easily synthesized, and the RAFT polymerization of styrene with this CTA afforded poly(N-octylbenzamide)-block polystyrene with controlled molecular weight and narrow polydispersity. PMID- 21638400 TI - Stepwise Self-Assembly of P3HT/CdSe Hybrid Nanowires with Enhanced Photoconductivity. AB - A facile approach to prepare poly(3-hexylthiophene) (P3HT)/cadmium selenide quantum dot (CdSe QD) hybrid coaxial nanowires by a stepwise self-assembly process is reported. P3HT nanowires of ~20 nm diameter are first prepared by self assembly in a poor solvent such as cyclohexanone, and then as-prepared CdSe QDs are deposited compactly onto the P3HT nanowires by non-covalent interactions between P3HT and CdSe. When illuminated with white light, the hybrid nanowires show enhanced photoconductivity compared with the pristine P3HT nanowires and the blended nanocomposites. PMID- 21638401 TI - Microwave-assisted one-pot synthesis of hyperbranched epoxide-amine adducts. AB - Hyperbranched epoxide-amine adducts were synthesized by a one-pot microwave (MW) assisted reaction. 4-(2,3-epoxypropyl-1-oxy)benzonitrile was hydrogenated using Pd/C under microwave conditions, forming the AB(2) monomer 1-aminomethyl-4-(2,3 epoxypropyl-1-oxy)benzene. Depending on the reaction temperature this monomer immediately reacts to give hyperbranched epoxide-amine adducts. Mass spectrometric investigations proved the formation of a homologous series of oligomers containing up to six repeating units. Due to the complexing properties of the poly(amino alcohol) moieties in the presence of Cu(2+) ions, large aggregates were formed. PMID- 21638403 TI - Macromol. Rapid commun. 17/2009. PMID- 21638402 TI - Macromol. Rapid commun. 17/2009. PMID- 21638404 TI - Novel nanostructures from self-assembly of chiral block copolymers. AB - A diblock copolymer system constituting both achiral and chiral blocks, polystyrene-block-poly(L-lactide) (PS-PLLA), was designed for the examination of chiral effects on the self-assembly of block copolymers (BCPs). A unique phase with three-dimensional hexagonally packed PLLA helices in PS matrix, a helical phase (H*), can be obtained from the self-assembly of PS-rich PS-PLLA with volume fraction of PLLA f PLLAv = 0.34, whereas no such phase was found in racemic polystyrene-block-poly(D.L-lactide) (PS-PLA) BCPs. Moreover, various interesting crystalline PS-PLLA nanostructures can be obtained by controlling the crystallization temperature of PLLA (T(c,PLLA) ), leading to the formation of crystalline helices (PLLA crystallization directed by helical confined microdomain) and crystalline cylinders (phase transformation of helical nanostructure dictated by crystallization) when T(c,PLLA) < T(g,PS) (the glass transition temperature of PS) and T(c,PLLA) ? T(g,PS) , respectively. As a result, a spring-like behavior of the helical nanostructure can be driven by crystallization so as to dictate the transformation (i.e., stretching) of helices and to result in crystalline cylinders. For PS-PLLA with PLLA-rich fraction (f PLLAv = 0.65), another unique phase, a hexagonally packed core-shell cylinder phase with helical sense (CS*), in which the PS microdomains appear as shells and PLLA microdomains appear as matrix and cores, can be found in the self-assembly of PLLA-rich PS-PLLA BCPs. The formation of those novel phases: helix and core shell cylinder is attributed to the chiral effect on the self-assembly of BCPs, so we named this PS-PLLA BCP as chiral BCP (BCP*). For potential applications of those materials, the spring-like behavior with thermal reversibility might provide a method for the design of switchable nanodevices, such as nanoscale actuators. In addition, the PLLA blocks can be hydrolyzed. After hydrolysis, helical nanoporous PS bulk and PS tubular texture can be obtained and used as templates for the formation of nanocomposites. PMID- 21638405 TI - Synthesis of Dendronized Diblock Copolymers via Click Chemistry: The Effect of Dendronization on Phase Separation Behaviour. AB - A novel route towards the synthesis of well-defined linear-dendronized diblock copolymers is reported. Precursor alkyne containing diblock copolymers were modified in a highly efficient cycloaddition reaction with dendritic azides of different generation. The dendronization has been shown to be selective and could be driven to completion under ambient conditions. The phase separation of such dendronized diblock copolymers was investigated in dependence of the generation size being attached. Compared to a linear-linear diblock copolymer as starting material the dendronization yielded in a pronounced phase separation. The nanoscaled features observed in thin films strongly depended on the dendron size and a variety of morphologies could be identified. Hence, the unique combination of controlled radical polymerization and click chemistry allows for the triggering of structured surfaces in the nanometer-regime. PMID- 21638406 TI - Defined Poly[styrene-block-(ferrocenylmethyl methacrylate)] Diblock Copolymers via Living Anionic Polymerization. AB - Ferrocenylmethyl methacrylate (FMMA) is one of the very few metallocene-based monomers that are promising candidates for truly living anionic polymerization. Nevertheless, FMMA homopolymers with a narrow polydispersity, or block copolymerization studies that result in satisfying blocking efficiencies, are unknown so far. Here we describe a procedure that leads to highly regular FMMA based polymers for the first time, characterized by polydispersity indices (PDI) of less that 1.05 and very high blocking efficiencies (>95%) in sequential copolymerization with styrene. Some of the obtained poly[styrene-block (ferrocenylmethyl methacrylate)]s show unusual microphase morphologies, presumably the consequence of high T(g) s causing 'frustrated' non-equilibrium states. PMID- 21638407 TI - Kinetics of Complex Formation between DNA and Cationically Charged Cylindrical Brush Polymers Observed by Stopped Flow Light Scattering. AB - The complex formation of pUC19 DNA and a cylindrical brush polymer with quaternized poly(vinyl pyridinium) side chains is investigated by stopped flow light scattering (LS). In highly dilute solutions up to two kinetic processes are observable, depending on the mole fraction of anionic charges, x(anion) . For x(anion) < 0.2 and x(anion) > 0.4-0.5 only one kinetic process was identified leading to the largest complexes at x(anion) = 0.4 well below charge stoichiometry. For 0.2 < x(anion) < 0.4 two kinetic growth processes were identified: The initially formed complexes on the time scale of a few hundred milliseconds keep growing by an unusual fractal growth process until after several minutes they become stable. This last fractal process does not follow the classical Smoluchowski coagulation theory as monitored by LS up to 90 s after mixing. PMID- 21638408 TI - Poly(3,4-ethylenedioxythiophene) Derived from Poly(ionic liquid) for the Use as Hole-Injecting Material in Organic Light-Emitting Diodes. AB - We report that poly(3,4-ethylenedioxythiophene) derived from poly(ionic liquid) (PEDOT:PIL) constitutes a unique polymeric hole-injecting material capable of improving device lifetime in organic light-emitting diodes (OLEDs). Imidazolium based poly(ionic liquid)s were engineered to impart non-acidic and non-aqueous properties to PEDOT without compromising any other properties of PEDOT. A fluorescent OLED was fabricated using PEDOT:PIL as a hole-injection layer and subjected to a performance evaluation test. In comparison with a control device using a conventional PEDOT-based material, the device with PEDOT:PIL was found to achieve a significant improvement in terms of device lifetime. This improvement was attributed to a lower indium content in the PEDOT:PIL layer, which can be also interpreted as the effective protection characteristics of PEDOT:PIL for indium extraction from the electrodes. PMID- 21638409 TI - An ionic molecular glass as electron injection layer for efficient polymer light emitting diode. AB - An ionic molecular glass based on a dendronized monoammonium salt has been facilely synthesized and utilized as an interfacial electron-injection layer in a light-emitting diode (LED). The characterization of a yellow-green LED that involves an Al cathode and a thin layer of the new compound spin cast from a methanol solution has shown device performances comparable to those obtained with a Ba/Al cathode. Photovoltaic measurements under white light irradiation reveal that a thin layer of the new compound can significantly increase the built-in potential and thus facilitate electron injection from an Al cathode. Furthermore, it is interesting to observe that the new ionic salt could undergo reorganization on the emissive conjugated polymer layer, which leads to the formation of nearly uniform nanoaggregates. PMID- 21638410 TI - Thermosensitive and dissolution properties in nanocomposite polymer hydrogels. AB - We investigate the phase transition behavior and dissolution resistant properties of thermo-sensitive nanocomposite hydrogels made from PEO-PPO-PEO triblock copolymer (Pluronic F127) and Laponite silicate nanoparticles. The rapid dissolution properties of F127 copolymer hydrogels usually limit their use as sustained release drug carriers. We overcome this limitation by synergistic combination of nanoparticle gelation characteristics with polymer thermo sensitivity. We present a proof of concept that the temperature-dependent phase transitions can be shifted as a function of hydrogel composition and that the dissolution of the polymer hydrogels as well as the release of a model drug, albumin, can be significantly slowed down by addition of nanoparticles. The dissolution resistant properties generated will prove useful in the future formulation, processing and application of our polymer hydrogels for sustained release drug delivery carriers. PMID- 21638411 TI - Self-Assembly of Nano Hydroxyapatite or Aragonite Induced by Molecular Recognition to Soy Globulin 7S or 11S. AB - Molecular self-assembly is emerging as a viable 'bottom-up' approach to build stable organic/inorganic nanometer-scale blocks. Herein, under the conditions of appropriate pH and ionic strength, soy globulin 7S or 11S were coprecipitated with hydroxyapatite (HAp) or aragonite (Arag), respectively, to fabricate two organic/inorganic hybrids: 7S/HAp and 11S/Arag. Results from high-resolution transmission electron microscopy show that the hybrids exhibit a nanosized core shell structure with globulin monomer 7S or 11S as core and HAp or Arag as shells. 7S/HAp and 11S/Arag present a disk and hexagon shape, respectively. After calcinations, monodispersed HAp without support from globulins existed as nanospheres. It was revealed that the globulin as host induces the self-assembly and growth layer by layer of HAp or Arag nanocrystals. The factors of molecular recognition and surface potential definitely affected the size and shape of the hierarchical blocks. This work provided a novel pathway to controllably synthesize a wide variety of precise plant protein/biomineral hybrid biomaterials. PMID- 21638412 TI - An efficient and environmentally friendly method for the synthesis of cellulose carbamate by microwave heating. AB - An efficient, solvent-free, and catalyst-free microwave synthesis of cellulose carbamate from a mixture of native cellulose and urea is reported. The structure of the samples are characterized by elemental analysis, FT-IR spectroscopy, X-ray diffraction, and NMR spectrometry. Cellulose carbamates with a nitrogen content of 0.651-2.427% are obtained by microwave heating at 255 W for 2-5 min. With the introduction of carbamate groups, the hydrogen bonding and crystalline structure of the native cellulose are partially destroyed. The products retain the cellulose I crystalline form of the native cellulose, and display good solubility in NaOH solutions. PMID- 21638413 TI - Microphase-Separated Brushes on Square Platelets in PS-b-PEO Thin Films. AB - In polystyrene-block-poly(ethylene oxide) thin films, microphase-separated brushes on the square platelets can be obtained via fast solvent evaporation by controlling the tethering density (0.08 < sigma < 0.11). The tethering density of the brushes is proportional to the thickness of the PEO crystal and increases with increasing initial solution heating temperature (T(i) ). When T(i) < T(m) , where T(m) is the melting point of PEO, brushes with microphase-separated structures are observed. The formation of microphase-separated brushes depends on two factors: the strong incompatibility between PS and noncrystalline PEO chains (attached to the crystalline PEO) and the weak interaction between PS-PS brushes. PMID- 21638414 TI - Macromol. Rapid commun. 18/2009. PMID- 21638415 TI - Supramolecular self-assembly of nonlinear amphiphilic and double hydrophilic block copolymers in aqueous solutions. AB - Supramolecular self-assembly of block copolymers in aqueous solution has received ever-increasing interest over the past few decades due to diverse biological and technological applications in drug delivery, imaging, sensing and catalysis. In addition to relative block lengths, molecular weights and solution conditions, chain architectures of block copolymers can also dramatically affect their self assembling properties in selective solvents. This feature article mainly focuses on recent developments in the field of supramolecular self-assembly of amphiphilic and double hydrophilic block copolymers (DHBCs) possessing nonlinear chain topologies, including miktoarm star polymers, dendritic-linear block copolymers, cyclic block copolymers and comb-shaped copolymer brushes. PMID- 21638416 TI - A Triphenylamine-Based Conjugated Polymer with Donor-pi-Acceptor Architecture as Organic Sensitizer for Dye-Sensitized Solar Cells. AB - A conjugated polymer containing an electron donating backbone (triphenylamine) and an electron accepting side chain (cyanoacetic acid) with conjugated thiophene units as the linkers has been synthesized. Dye-sensitized solar cells (DSSCs) are fabricated utilizing this material as the dye sensitizer, resulting a typical power conversion efficiency of 3.39% under AM 1.5 G illumination, which represents the highest efficiency for polymer dye-sensitized DSSCs reported so far. The results show the good promise of conjugated polymers as sensitizers for DSSC applications. PMID- 21638417 TI - Layer-by-Layer Assembled Nanotubes as Biomimetic Nanoreactors for Calcium Carbonate Deposition. AB - Enzyme-loaded magnetic polyelectrolyte multilayer nanotubes prepared by layer-by layer assembly combined with the porous template could be used as biomimetic nanoreactors. It is demonstrated that calcium carbonate can be biomimetically synthesized inside the cavities of the polyelectrolyte nanotubes by the catalysis of urease, and the size of the calcium carbonate precipitates was controlled by the cavity dimensions. The metastable structure of the calcium carbonate precipitates inside the nanotubes was protected by the outer shell of the polyelectrolyte multilayers. These features may allow polyelectrolyte nanotubes to be applied in the fields of nanomaterials synthesis, controlled release, and drug delivery. PMID- 21638418 TI - Discriminating Among Co-monomer Sequence Distributions in Random Copolymers Using Interaction Chromatography. AB - Interaction chromatography has been employed to validate that adsorption of poly[styrene-co-(4-bromostyrene)] (PBr(x) S) random copolymers, where x denotes the mole fraction of 4-bromostyrene (4-BrS) in PBr(x) S in solution depends on the average number of adsorptive segments, the type of adsorbing substrate, and on the co-monomer sequence distribution in PBr(x) S. PMID- 21638419 TI - Substrate-induced controllable wrinkling for facile nanofabrication. AB - In this work, we report a facile and effective strategy to generate patterned wrinkles. This strategy includes first adhering a thin poly(dimethylsiloxane) (PDMS) film (<82 um) on porous conductive adhesive tape (CAT), followed by sputter coating of Au onto PDMS under vacuum condition, which results in formation of patterned wrinkles on the Au-PDMS bilayer. CAT was found to induce local stretching of PDMS thin film, which was the key for controlled wrinkle formation. Compared with previous wrinkling methods, our strategy is simpler and gives smaller feature sizes (down to 300 nm). PMID- 21638420 TI - Polymerizable ionic liquid crystals. AB - Polymerizable vinylimidazolium ionic liquids (ILs) that contain mesogenic coumarin and biphenyl units, respectively, have been synthesized. The N alkylation of N-vinylimidazole with bromoalkylated mesogenic units 7-(6 bromohexyloxy)coumarin (1) and 4,4'-bis(6-bromohexyloxy)biphenyl (2) was then carried out. The thermal behavior of the obtained ILs 3 and 4 was investigated by differential scanning calorimetry and polarizing optical microscopy. These measurements showed that the attached mesogenic units induce the self-assembly of ILs and, therefore, the occurrence of liquid crystalline phases. Subsequently, the ionic liquid crystals (ILCs) 3 and 4 were polymerized by a free-radical mechanism. PMID- 21638421 TI - Amphiphilic Hybrid pi-Conjugated Polymers Containing Polyhedral Oligomeric Silsesquioxanes. AB - Amphiphilic hybrid pi-conjugated polymers that have polyhedral oligomeric silsesquioxanes on their side chains have been successfully synthesized by the Sonogashira-Hagihara polycondensation reaction. The obtained polymers were studied with ultraviolet-visible absorption and photoluminescence spectra. In these polymers, the pi-conjugation length was extended along the poly(p-phenylene ethynylene) backbone. Furthermore, the content of the POSS substituents can influence the aggregation behavior of the polymers and subsequent luminescent properties. PMID- 21638422 TI - Reactive hydrogel networks for the fabrication of metal-polymer nanocomposites. AB - In this study, highly stable gold and silver nanoparticles evenly distributed within a crosslinked poly(acrylamide)/poly(N-(hydroxymethyl)acrylamide) (PAAm PHMAAm) network have been fabricated without addition of a reducing agent. Remarkably, the same chemical hydrogel composition has been involved in the successful fabrication of spherical gold and silver nanoparticles within the hydrogel template. The hydrogel network acts simultaneously as an efficient reducing agent and stabilizer. The PAAm-PHMAAm hydrogel network binds metal ions and, following reduction of bound to crosslinked template metal ions, proceeds via oxidation of hydroxymethyl hydrogel fragments. A one-electron mechanism is proposed for the formation of the silver and gold nanoparticles. PMID- 21638423 TI - Cross-linked conjugated polymers for achieving patterned three-color and blue polymer light-emitting diodes with multi-layer structures. AB - Reactions between the ethylene groups in the backbone of conjugated polymers under UV illumination and heat treatment result in the cross-linking of the main polymer chains. The cross-linking leads to two simultaneous results in the polymer: excellent solvent resistance and increased bandgap. Using this reaction, three-color polymer light-emitting diodes (PLEDs) with a multi-layer structure can be easily realized by a dry photo-pattern in an active-gas-free environment. Multi-layer blue devices with dramatically enhanced efficiency can also be achieved conveniently. PMID- 21638424 TI - A novel strategy for the synthesis of sheet-like polyaniline. AB - Polyaniline (PANI) micro/nanosheets are successfully synthesized by a template free method without using any conventional oxidants. Scanning electron microscopy, transmission electron microscopy, and FT-IR spectroscopy are applied to characterize the products. By investigating the morphologies and chemical structures of the PANI micro/nanosheets, a possible formation mechanism is proposed. In addition, the influences of experimental parameters, such as the kind of dopant, concentration of aniline, and acidity of reaction system, on the morphologies of the PANI micro/nanosheets have been systematically investigated. PMID- 21638425 TI - Preparation of ag-embedded polystyrene nanospheres and nanocapsules by miniemulsion polymerization. AB - An isopropyl myristate (IPM) biocompatible oil and an IPM solution of dodecanethiol-capped Ag nanoparticles (NPs, 4.5 nm) were used as hydrophobes to suppress the Ostwald ripening of monomer/hydrophobe miniemulsified droplets in a surfactant-stabilized water phase. The formation of non-IPM-encapsulated nanospheres (48 nm) and IPM-encapsulated nanocapsules (90 nm) were precisely controlled by using a water-soluble and an oil-soluble initiator, respectively, in the presence of a pure IPM as a hydrophobe in miniemulsion polymerization. Well-defined PS nanospheres, on which surfaces were coated with Ag NPs (Ag/PS nanospheres, 65 nm), and nanocapsules encapsulating both NPs and IPM liquid phase (Ag-IPM/PS nanocapsules, 115 nm) were made by replacing the hydrophobe from pure IPM with Ag/IPM solution. These nanostructures were characterized by transmission and scanning electron microscopes. PMID- 21638426 TI - Thinner is Better: An Ultrathin Conducting Oligoaniline Film for Gas Microsensors with Ultralow Detection Limits. AB - An ultralow-limit gas microsensor based on an ultrathin conducting oligoaniline film integrated with microscale gold electrodes is developed. A nanoscale oligoaniline film is fabricated on a poly(dimethylsiloxane) (PDMS) substrate using graft polymerization using FeCl(3) , a mild oxidant, rather than conventional (NH(4) )(2) S(2) O(8) . The as-fabricated film is around 14 nm in thickness and above 85% transmittance on a PDMS substrate with a smooth surface morphology and high conductivity. Taking NH(3) as a protocol, the nanoscale oligoaniline film microsensor shows an ultralow detection limit to the ppb level with more rapid response and high sensitivity to NH(3) compared to the thicker PANI film using conventional methods. PMID- 21638427 TI - Macromol. Rapid commun. 19/2009. PMID- 21638428 TI - Hairy Core-Shell Nanoparticles via RAFT: Where are the Opportunities and Where are the Problems and Challenges? AB - The preparation of hairy core-shell nanoparticles including (crosslinked) micelles, unimolecular micelles such as star polymers with block structures in each arm and surface grafted nanoparticles such as inorganic particles via the RAFT process are discussed. The RAFT process is certainly a highly versatile process. However, it should not be forgotten that RAFT polymerization is a process, i.e., superimposed on a conventional free radical process. Furthermore, the livingness of the process is dependent on the accessibility of the RAFT group, which can be hampered in certain approaches such as star synthesis and surface grafting from nanoparticles. Nevertheless, the RAFT process is a versatile toolbox that offers good solutions to a range of problems in the preparation of hairy nanoparticles. PMID- 21638429 TI - On the quantitative click conjugation of molecular weight distributions: what can theoretically be expected? AB - The assessment of the success of a click conjugation for block copolymer formation often occurs via ill-defined criteria based on the qualitative shape of the size exclusion chromatography (SEC) distribution as well as unsuitable characteristics of these molecular weight distribution such as the peak molecular weight, M(p) . The data presented herein illustrate that from the shape of the SEC distribution, w(log M) versus log M, of the click product-under the assumption of a 100% efficient conjugation reaction-a conclusion about the effectiveness of the click conjugation cannot be derived. It is demonstrated that under certain conditions multi-modal molecular weight distributions are obtained from two well-defined monomodal initial distributions. Similarly, the comparison of peak maxima between the conjugate and the initial SEC distributions is a poorly defined criterion to assess the success of a conjugation. The only reliable assessment for the success of the click coupling reaction is the true number average molecular weight, $?overline M _{?rm n}$, of the SEC distribution of the click product, as well as a plot of the concentration or number distributions, c versus M, of the precursors and product polymer. PMID- 21638430 TI - Three-dimensional polycaprolactone hierarchical scaffolds supplemented with natural biomaterials to enhance mesenchymal stem cell proliferation. AB - A hybrid technology that combines a three-dimensional (3-D) dispensing system with an electrospinning process was used to produce a hierarchical 3-D scaffold consisting of micro-sized polycaprolactone (PCL) strands and micro/nano-sized fibres. The micro/nanofibre biocomposites electrospun with PCL/small intestine submucosa (SIS) and PCL/Silk fibroin were layered between melt-plotted micro strands. The scaffold containing SIS exhibited a stronger hydrophilic property than other scaffolds due to the various hydrophilic components in SIS. The 3-D hierarchical scaffold having biocomposites exhibited an incredibly enhanced initial cell attachment and proliferation of bone marrow-derived mesenchymal stem cells relative to the normally designed 3-D scaffold. PMID- 21638431 TI - Micropatterned fiber scaffolds for spatially controlled cell adhesion. AB - Because the local microstructure plays a pivotal role for many biological functions, a wide range of methods have been developed to design precisely engineered substrates for both fundamental biological studies and biotechnological applications. However, these techniques have been by-and-large limited to flat surfaces. Herein, we use electrohydrodynamic co-spinning to prepare biodegradable three-dimensional fiber scaffolds with precisely engineered, micrometre-scale patterns, wherein each fiber is comprised of two distinguishable compartments. When bicompartmental fiber scaffolds are modified via spatially controlled peptide immobilization, highly selective cell guidance at spatial resolutions (<10 um), so far exclusively reserved for flat substrates, is achieved. Microstructured fiber scaffolds may have utility for a range of biotechnological applications including tissue engineering or cell-based assays. PMID- 21638432 TI - Combinatorial Energy Transfer between an End-Capped Conjugated Polyelectrolyte and Chromophore-Labeled PNA for Strand-Specific DNA Detection. AB - We report a macromolecular end-capping approach to improve the detection sensitivity of cationic conjugated polymer (CCP) based DNA detection. A phenylethynyl anthracene (PEA) end-capped cationic polyfluorene (PF) derivative (P1) is synthesized via Suzuki coupling. Due to efficient fluorescence resonance energy transfer (FRET) from the polymer backbone to the end-capper PEA units, the polymer (P1) fluorescence is dominated by the emission from PEA even in dilute aqueous solution. P1 emission has a better spectral overlap with fluorescein (Fl) absorption compared to that for uncapped PF (P2). In addition, the intra and intermolecular energy transfer for P1 is more efficient in the presence of DNA due to complexation-induced polymer aggregation. These impart a combinatorial FRET between P1 and an Fl-labeled probe which is more efficient than that between P2 and the same probe. P1 thus offers a better DNA detection sensitivity relative to P2 and opens up new opportunities to improve the performance of CCP based biosensors involving FRET. PMID- 21638433 TI - Glucosamine hydrochloride functionalized water-soluble conjugated polyfluorene: synthesis, characterization, and interactions with DNA. AB - Novel glucosamine hydrochloride functionalized water-soluble conjugated polyfluorene was easily synthesized through Cu(I)-catalyzed azide/alkyne "click" ligation and Suzuki coupling polymerization. The water-solubility and biocompatibility of the polymer were improved after grafting glucosamine hydrochloride to the side chains of the conjugated polymer. As a fluorescent model system of chitosan, its interaction with single-stranded DNA was studied by spectrofluorometric titration. PMID- 21638434 TI - Damping behavior of the aggregation-disaggregation self-oscillation of a polymer chain. PMID- 21638435 TI - Polyaniline "nanotube" self-assembly: the stage of granular agglomeration on nanorod templates. AB - The identification and control of a critical stage of polyaniline "nanotube" self assembly is presented, namely the granular agglomeration or growth onto nanorod templates. When the synthesis pH is held above 2.5, smooth insulating nanorods exhibiting hydrogen bonding and containing phenazine structures are produced, while below pH 2.5, small 15-30 nm granular polyaniline nanoparticles appear to agglomerate onto the available nanorod surface, apparently improving conductivity of the resulting structures by three orders of magnitude. This finding affects both fundamental theories of polyaniline nanostructure self-assembly and their practical applications. PMID- 21638436 TI - pH Tailoring Electrical and Mechanical Behavior of Polymer-Clay-Nanotube Aerogels. AB - Aerogels are low density (<0.1 g . cm(-3) ), highly porous materials that are especially interesting for insulating applications. Combinations of clay and water-soluble polymers are commonly used to produce aerogels, but these materials are often mechanically weak. Single-walled carbon nanotubes (SWNT) were combined with clay and found to significantly improve mechanical behavior and impart electrical conductivity to these aerogels. Poly(acrylic acid) (PAA) as the matrix polymer provides a means of tailoring the electrical conductivity and mechanical behavior by altering the pH of the aqueous aerogel precursor suspensions prior to freeze drying. An aerogel, made from a pH 9 aqueous suspension containing 0.5 wt. % PAA, 5 wt.-% clay, and 0.05 wt.-% SWNT, has a compressive modulus of 373 kPa. In the absence of nanotubes, this modulus is reduced to 43 kPa. Reducing suspension pH to 3, prior to freeze drying, also reduces modulus for these aerogels, but electrical conductivity is increased when nanotubes are present. It was found that bundled nanotubes provide better reinforcement for these low density composites, which may provide some new insight into the use of nanotubes in materials that will be exposed to compressive loading. PMID- 21638437 TI - Controlling orientation and functionalization in thin films of block copolymers. AB - Functionalizing and controlling nanostructures resulting from block copolymer self-assembly are key factors in defining their application. In this work, a simple but quite general route to achieve both goals simultaneously is discussed. In thin films of polystyrene-block-poly(vinyl pyridine) (PS-b-PVP) with small concentrations of a gold salt, the salt is found to complex with the PVP block which leads to an orientation of the microdomains normal to the surface after solvent annealing together with functionalization. By increasing the amount of gold salt, on the other hand, micelles are found to form in solutions leading to a range of different morphologies in the thin films. PMID- 21638439 TI - Macromol. Rapid commun. 20/2009. PMID- 21638438 TI - Synthesis of CdSe-Poly(N-vinylcarbazole) Nanocomposite by Atom Transfer Radical Polymerization for Potential Optoelectronic Applications. AB - A hybrid inorganic-polymer nanocomposite using CdSe nanocrystals with high electron mobility has been successfully synthesized by atom transfer radical polymerization (ATRP). First the hydroxyl-coated CdSe nanoparticles (i.e., CdSe OH) were prepared via a wet chemical route. A polymerization initiator was then prepared for ATRP of N-vinylcarbazole. FT-IR, (1) H NMR, and XRD analyses confirmed the successful synthesis of CdSe-poly(N-vinylcarbazole) (PVK) nanohybrid. UV-Vis spectra and photoluminescence data revealed that grafting of PVK onto the surface of CdSe nanocrystals would reduce the band gap of PVK and cause the red shift of emission peak. TEM and SEM micrographs exhibited CdSe nanoparticles that were well-coated with PVK polymer. PMID- 21638440 TI - High-throughput screening in olefin-polymerization catalysis: from serendipitous discovery towards rational understanding. AB - High-throughput-screening (HTS) tools and methods are used more and more, especially in industry, in the search for new, selective organometallic catalysts. In most cases, the approach is, in essence, empirical, and the strategy is to increase the number of experiments that can be run at a given place in a given time. Highly miniaturized, parallel reaction setups have been implemented for the rapid assessment of whether novel catalysts resulting from the structural amplification of a basic framework are "good" or "bad" with respect to the properties of interest, and, depending on the response, worthy of a subsequent, more-careful evaluation. In this article, we demonstrate that it is possible to utilize these state-of-the-art HTS platforms with a different strategy: the rapid generation of reliable kinetic data for mechanistic studies in view of a thorough understanding and rational catalyst design. Ziegler-Natta type catalytic olefin polymerization will be used throughout as an example. PMID- 21638441 TI - New light on old wisdoms on molten polymers: conformation, slippage and shear banding in sheared entangled and unentangled melts. AB - The flow of viscoelastic materials is usually interpreted as resulting from intramolecular properties. Typically, the non-linear flow behaviour and sluggish relaxation dynamics in entangled polymers are interpreted by a disentanglement process. This molecular interpretation has never been validated by direct observation. We report here on in situ observations of polymer melts under steady state shear flow using neutron scattering and particle tracking velocimetry. It is shown that the chains remain largely undeformed under steady-state shear flow whereas wall slippage and shear-banding are identified in both entangled and unentangled polymer melts. These observations are of prime importance; they reveal that the flow mechanism and its viscoelastic signature reflect a collective effect and not properties of individual chains. PMID- 21638442 TI - Polymer Fluoride Sensors Synthesized by RAFT Polymerization. AB - A series of polymer chemosensors containing naphthalimide signal moiety and imide recognition moiety for the selective detection of fluoride ions (F(-) ) were synthesized by reversible addition-fragmentation chain transfer (RAFT) polymerization. The polymer chemosensors display obvious absorption and fluorescence variations upon the addition of F(-) , which can be observed by both naked eyes and optical responses. The polymers showed higher fluorescence enhancement than its monomer, and the polymers with higher molecular weight have higher sensitivity than those with lower molecular weight. Other halide anions are found to hardly induce any variation of either the absorption or fluorescence spectra. PMID- 21638443 TI - Color-oscillating photonic crystal hydrogel. AB - In this work, a color-oscillating system is first developed by combining the intrinsic peristaltic motion of a Landolt pH-oscillator with the structure color of a well-designed pH-sensitive photonic crystal hydrogel. As a result, the pH oscillation reaction procedure could be distinctly monitored by the distinct change of structure color/optic signal. The oscillation rhythm of the pH well coincides with that of the stopband/structure color. The oscillation detail of each cycle can also be clearly monitored by color change. This work would be of great significance for the promising applications of real-time monitoring of the microactuator by optical signal or structure color. PMID- 21638444 TI - Precise control of photoinduced birefringence in azobenzene-containing liquid crystalline polymers by post functionalization. AB - A series of functionalized liquid-crystalline polymer materials with different degrees of functionality was synthesized by a post Sonogashira cross-coupling reaction of a polymer precursor. The post-functionalization was carried out under mild conditions and showed a high yield. Although a highly birefringent azotolane group was introduced into the polymer precursor, the photoresponse of the functionalized liquid-crystalline materials was not obviously changed. By adjusting the content of azotolane groups, precise control of the photoinduced birefringence was successfully obtained after thermal enhancement upon annealing. The present method to gain precise control of photoinduced birefringence might enable one to finely photocontrol the optical performances of materials, and may have a potential application as an advanced process for photonic materials. PMID- 21638445 TI - Metal-free activation in the anionic ring-opening polymerization of cyclopropane derivatives. AB - The successful activation observed when using Bu(t) P(4) phosphazene base and thiophenol or bisthiols for the anionic ring opening polymerization (ROP) of di-n propyl cyclopropane-1,1-dicarboxylate is described. Well-defined monofunctional or difunctional polymers with a very narrow molecular weight distribution were obtained through a living process. Quantitative end-capping of the propagating malonate carbanion was accessible by using either an electrophilic reagent such as allyl bromide or a strong acid such as HCl. Kinetics studies demonstrated a much higher reactivity compared to the conventional route using alkali metal thiophenolates. PMID- 21638446 TI - 3-Ethylated N-Vinyl-2-pyrrolidone with LCST Properties in Water. AB - The monomer 3-ethyl-1-vinyl-2-pyrrolidone (3) and the homopolymer poly(3-ethyl-1 vinyl-2-pyrrolidone) (5) have been synthesized. Polymer 5 is soluble in water and shows a critical temperature (T(c) ) of 27 degrees C. The presence of cyclodextrin causes a slight shift of the T(c) . The lower critical solution temperature (LCST) could be varied between 27 and 40 degrees C by copolymerization with N-vinyl-2-pyrrolidone. A linear correlation between the T(c) and the copolymer composition is observed. PMID- 21638447 TI - Macromolecular Recognition of Cyclodextrin: Inversion of Selectivity of beta Cyclodextrin toward Adamantyl Groups Induced by Macromolecular Chains. AB - Interaction of beta-cyclodextrin (beta-CD) with alternating copolymers (pAdMA and pAdPhMA) of sodium maleate with adamantyl (Ad) and with adamantylphenyl (AdPh) vinyl ether has been investigated by several NMR techniques. Comparing the apparent association constants (K) for the polymers with the K for the model compounds, which are determined by the analysis of (1) H NMR and isothermal titration calorimetry data, respectively, the selectivities of beta-CD toward Ad and AdPh moieties are contrasting for the model and polymer systems. This phenomenon is described by circular dichroism and 2D NMR as follows; the most stable inclusion complex for the beta-CD/AdPh model system is extremely destabilized for the corresponding polymer system because of competition with hydrophobic interaction between neighboring AdPh moieties. PMID- 21638448 TI - Synthesis of Dendritic Oligo-Spiro(fluorene-9,9'-xanthene) Derivatives with Carbazole and Fluorene Pendants and their Thermal, Optical, and Electroluminescent Properties. AB - Two novel spiro-configured ter(arylene-ethynylene) derivatives, TSF-Cz and TSF-F, have been designed and synthesized using spiro(fluorene-9,9'-xanthene) (SFX) as building blocks, introducing a hole-transporting carbazole and a fluorene chromophore as the peripheral functional group into the backbone through an oxygen atom. The two well-defined oligomers possess good solubility, film-forming quality, and high T(g) 's at 140 and 126 degrees C, respectively. In addition, these oligomers exhibit blue photoluminescence (PL) emission both in solution and solid states. The double-layered devices fabricated using the two materials as the emitter show a sky-blue emission with a brightness and a current efficiency of 7 613 cd . m(-2) and 1.11 cd . A(-1) for TSF-Cz, and 1 507 cd . m(-2) and 0.36 cd . A(-1) for TSF-F, respectively. PMID- 21638449 TI - Nanoscale Networks in Poly[isoprene-block-styrene-block-(methyl methacrylate)] Triblock Copolymers. PMID- 21638450 TI - Controlled stripping of polyelectrolyte multilayers by quaternary ammonium surfactants. AB - The quartz crystal microbalance with dissipation technique (QCM-D) and atomic force microscopy (AFM) have been employed to study the interaction of N tetradecyl trimethyl ammonium bromide (TdTmAB) with polyelectrolyte multilayers containing poly(sodium 4-styrene sulfonate) (PSS) as the polyanion and either poly(allylamine hydrochloride) (PAH) or poly(diallyl dimethyl ammonium chloride) (PDADMAC) as the polycations. The multilayers were exposed to aqueous solutions of TdTmAB. This resulted in a selective removal of PDADMAC PSS layers while layers with PAH as polycation remained stable. It is suggested that PDADMAC/PSS multilayers can be employed as strippable protecting layers. PMID- 21638451 TI - Novel highly active niobium catalysts for ring opening metathesis polymerization of norbornene. AB - Ring-opening metathesis polymerization (ROMP) of norbornene catalyzed by niobium(V) N,N-dialkylcarbamates Nb(O(2) CNR(2) )(5) , R = Et (1), Me (2) was studied in the presence of methylaluminoxane (MAO) as a cocatalyst. These novel catalytic systems resulted very active in chlorobenzene: 1 in the presence of methylaluminoxane catalyzes the ROMP of norbornene with the highest activity (29 000 kg of polymer/mol of catalyst * hour) never reported up to now for niobium catalysts. The high productivity appears particularly attractive considering that these precursors are rather cheap and easy to synthesize and to handle. Polynorbornenes were characterized by FT-IR and NMR spectroscopies and by DSC calorimetry. A new FT-IR method for the swift determination of the cis/trans content of the polymer is presented. PMID- 21638453 TI - Macromol. Rapid commun. 21/2009. PMID- 21638452 TI - Fabrication of Nanostructured Self-Detoxifying Nanofiber Membranes that Contain Active Polymeric Functional Groups. AB - Military soldiers, medicinal doctors, and ordinary people require protection against chemical and biological warfare (C&B) agents. Activated charcoal impregnated with metal ions is currently used in protective clothing applications, which has some disadvantages. Electrospinning is emerging as one of the cheapest technologies to produce continuous nanofibers with a high surface area-to-volume ratio. In the present study, electrospinning of a poly(ethylene imine) (PEI)/nylon blend has been carried out in which PEI acts as a support material as well as a catalytic media. The membrane is combined with non selective metal oxide nanoparticles to degrade C&B agents into non-toxic products. In addition, these membranes possess hydrophilic properties, hence they are suitable candidates for protective clothing applications. PMID- 21638454 TI - Micellization phenomena in semicrystalline block copolymers: reflexive and critical views on the formation of cylindrical micelles. AB - Investigations on the self-assembly of block copolymers in solution have in some way a less well-studied history than the study of their phase separation in the solid state, and many aspects are yet not completely understood. Here we focus on the behavior of a specific class of copolymers, namely semicrystalline block copolymers, capable of forming cylindrical aggregates in a solvent selective for the non-crystalline, complementary block. A common model of micellization is proposed, in principle applicable to most of these copolymeric systems. PMID- 21638455 TI - Ultra-Fast RAFT-HDA Click Conjugation: An Efficient Route to High Molecular Weight Block Copolymers. AB - The use of the reversible addition fragmentation chain transfer-hetero Diels Alder (RAFT-HDA) click reaction for the modular construction of block copolymers is extended to the generation of high molecular weight materials. Cyclopentadienyl end-functionalized polystyrene (PS-Cp) prepared via both atom transfer radical polymerization (ATRP) and the RAFT process are conjugated to poly(isobornyl acrylate) (PiBoA) (also prepared via RAFT polymerization) to achieve well-defined block copolymers with molecular weights ranging from 34 000 to over 100 000 g . mol(-1) and with small polydispersities (PDI < 1.2). The conjugation reactions proceeded in a very rapid fashion (less than 10 min in the majority of cases) under ambient conditions of temperature and atmosphere. The present study demonstrates-for the first time-that RAFT-HDA click chemistry can provide access to high molecular weight block copolymers in a simple and straight forward fashion. PMID- 21638456 TI - In situ Pressure Fluctuations of Polymer Melt Flow Instabilities: Experimental Evidence about their Origin and Dynamics. AB - Despite the practical importance of polymer melt instabilities, there is still a lack of experiments able to characterize in situ the origin and behavior of these phenomena. In this context, a new set-up consisting of high sensitive pressure transducers located inside a slit-die and an advanced mathematical framework to process in situ measurements of polymer melt instabilities, are developed and applied. Our results show for the first time that pressure oscillations can actually be detected inside the die under sharkskin conditions. This originates from a factor of 10(3) and 10(2) improvement in terms of time and pressure resolution. Furthermore, new evidence towards the propagation of the slip phenomena along the die in spurt instabilities are found. PMID- 21638457 TI - SP-PLP-EPR Investigations into the Chain-Length-Dependent Termination of Methyl Methacrylate Bulk Polymerization. AB - Termination kinetics of methyl methacrylate (MMA) bulk polymerization has been studied via the single pulsed laser polymerization-electron paramagnetic resonance method. MMA-d(8) has been investigated to enhance the signal-to-noise quality of microsecond time-resolved measurement of radical concentration. Chain length-dependent termination rate coefficients of radicals of identical size, k ti,i, are reported for 5-70 degrees C and up to i = 100. k ti,i decreases according to the power-law expression $k_{?rm t}^{i,i} = k_{?rm t}^{{?rm 1,1}} ?cdot i^{ - ?alpha }$. At 5 degrees C, k(t) for two MMA radicals of chain-length unity is k t1,1 = (5.8 +/- 1.3) . 10(8) L . mol(-1) . s(-1) . The associated activation energy and power-law exponent are: E(A) (k t1,1) ~ 9 +/- 2 kJ . mol( 1) and alpha ~ 0.63 +/- 0.15, respectively. PMID- 21638458 TI - Highly Active Syndiospecific Living Polymerization of Higher 1-Alkene with ansa Fluorenylamidodimethyltitanium Complex. AB - 1-Hexene polymerization was conducted by [t-BuNSiMe(2) (3,6-t-Bu(2) Flu)]TiMe(2) (1) using trialkylaluminum-free modified methylaluminoxane (dMMAO) as a cocatalyst in toluene. The system produced living syndiotactic poly(1-hexene) with high turnover frequency of propagation (TOF, 98 s(-1) ) at 0 degrees C. The propagation rate was increased linearly against the 1-hexene concentration, which indicates that the first-order dependence of the propagation rate on monomer concentration. Polymerizations of 1-octene, 1-decene and 1-dodecene were also conducted for investigating the effect of chain length of 1-alkene on propagation rate by means of the livingness of this system. The propagation rate decreased according to the chain length until 1-decene but almost unchanged in longer 1 alkene: TOF, 1-octene (62 s(-1) )>1-decene (32 s(-1) ) ~ 1-dodecene (31 s(-1) ). PMID- 21638459 TI - Photochemical generation of ferrocene-based redox-polymer networks. AB - A photochemical approach toward the generation of enzyme-containing redox polymer networks, which are the key material in enzymatic sensors and biofuel cells, is described. The approach is based on the incorporation of photo-reactive benzophenone groups into the redox polymers. The obtained polymers are then deposited on the surface of glassy carbon electrodes and cross-linked by illumination with UV light at 365 nm. If this step is done in the presence of the enzyme glucose oxidase, functional electrodes are obtained that yield electrical power upon addition of glucose. This work specifically addresses the question of electrode stability in buffer and demonstrates how slight variations in the chemistry of the redox polymer have a dramatic effect on the electrochemical performance of the electrodes. Different ferrocene-containing redox polymer networks are synthesized and their properties in physiological buffer are studied. PMID- 21638460 TI - Revealing triple-shape memory effect by polymer bilayers. AB - Bilayer polymers that consist of two epoxy dual-shape memory polymers of well separated glass transition temperatures have been synthesized. These bilayer epoxy samples exhibit a triple-shape memory effect (TSME) with shape fixities tailorable by changing the ratio between the two layers. The triple-shape fixities of the bilayer epoxy polymers can be explained by the balance of stress between the two layers. Based on this work, it is believed that the following three molecular design criterions should be considered in designing triple-shape memory polymers with optimum TSME: 1) well-separated thermal transitions, 2) a strong interface, and 3) an appropriate balance of moduli and relative ratios between the layers (or microphases). PMID- 21638461 TI - Plasma functionalization of multiwalled carbon nanotube bucky papers and the effect on properties of melt-mixed composites with polycarbonate. AB - Multiwalled carbon nanotubes in the form of bucky papers were modified using Ar/O(2) plasma and thereafter melt-mixed into polycarbonate. The effect of plasma modification on the nanotubes was followed by XPS, indicating the formation of carboxylic or ester groups at the nanotube surfaces. In the melt-mixed nanocomposites, the modified nanotubes exhibited a better macrodispersion and better phase adhesion to the matrix as evidenced by morphological investigations. The electrical percolation threshold was not altered and occurred below 0.5 wt.-% nanotubes. The mechanical properties were improved by having higher values of stress at yield, stress beyond the yield point, and strain at break illustrating the effect of both better dispersion and enhanced phase adhesion. PMID- 21638462 TI - Electrochemical Polymerisation of N-Arylated and N-Alkylated EDOT-Substituted Pyrrolo[3,4-c]pyrrole-1,4-dione (DPP) Derivatives: Influence of Substitution Pattern on Optical and Electronic Properties. AB - New pyrrolo[3,4-c]pyrrole-1,4-dione (DPP) derivatives carrying 3,4-ethylenedioxy thiophenylphenyl (EDOT-phenyl) substituent groups in the 3- and 6-position, or in the 2- and 5-position of the DPP chromophore were synthesised and electrochemically polymerised. The properties of the polymers were investigated using cyclic voltammetry and UV/Vis absorption spectroscopy. It was found that the optical and electronic properties differ greatly between the two polymers. Materials with EDOT-phenyl groups in the 3- and 6-positions represent conjugated polymers with a low oxidation potential and reversible electrochromic properties, whereas the polymer with EDOT-phenyl groups in the 2- and 5-positions is non conjugated and possesses a high oxidation potential and irreversible redox behaviour. PMID- 21638463 TI - One-Dimensional V(2) O(5) @Polyaniline Core/Shell Nanobelts Synthesized by an In situ Polymerization Method. AB - Uniform one-dimensional V(2) O(5) @polyaniline core/shell nanobelts have been fabricated by a simple in-situ polymerization method in the absence of any surfactant and additional initiator. The influences of pH and additional initiator on the morphology of the resulting products are investigated. The pH value is important for the formation of V(2) O(5) @polyaniline core/shell nanobelts, which preserve the original morphology of V(2) O(5) nanobelts. With a decrease in the pH value to 0 the original morphology of the V(2) O(5) nanobelts is destroyed. When ammonium peroxydisulfate is used, some separated polyaniline nanofibers are formed. The formation of the V(2) O(5) @polyaniline core/shell nanobelts can be related to the in-situ polymerization of aniline monomer by etching V(2) O(5) nanobelts. The electrochemical lithium intercalation/deintercalation of V(2) O(5) @polyaniline core/shell nanobelts is investigated by cyclic voltammograms. PMID- 21638464 TI - Influence of PEG-ran-PPG Surfactant on Vapour Phase Polymerised PEDOT Thin Films. AB - The oxidant, Fe(III) tosylate, was used in the vapour phase polymerisation (VPP) of PEDOT. The amphiphilic co-polymer poly(ethylene glycol-ran-propylene glycol) was added and its influence examined. Both the PEDOT conductivity and optical contrast range increased with the inclusion of the co-polymer, with the maximum being recorded at 4 wt.-%. Loadings higher than this resulted in a systematic decrease in both conductivity and optical contrast. Evidence indicates that in addition to the beneficial anti-crystallisation effect to the oxidant layer, the co-polymer also reduces the effective reactivity of the oxidant, as demonstrated by slower polymerisation rates. Confirmation of the change in polymerisation rate was obtained using a quartz crystal microbalance (QCM). The slower polymerisation rate results in higher conductivity and optical contrast; however, XPS data confirmed that the co-polymer remained within the PEDOT film post-washing and this result explains why the performance decreases at high surfactant loadings. PMID- 21638465 TI - Poly(sulfonated phenylene)-block-Polyimide Copolymers for Fuel Cell Applications. AB - Novel poly(2-(3-sulfo)benzoyl-1,4-phenylene)-block-polynaphthalimide (PSP-b-PI) copolymers were successfully synthesized by Ni(0)-catalyzed copolymerization of 2,5-dichloro-3'-sulfo-benzophenone and dichloro-terminated naphthalimide oligomer. The membranes exhibited a microphase-separated structure and good hydrolytic stability at 130 degrees C. They showed a fairly strong anisotropy of membrane swelling with much smaller in-plane swelling, but a rather weak anisotropy of proton conductivity. The membranes had a fairly high through-plane conductivity in water and even under low relative humidity. The PSP-b-PI copolymer with an IEC of 1.5 meq . g(-1) showed high PEFC performance due to the high through-plane conductivity. PMID- 21638466 TI - From nanodot to nanowire: hybrid au/titania nanoarrays by block copolymer templates. AB - A simple scheme to fabricate 2-D arrays of Au/titania hybrid nanopatterns is presented, using polystyrene-block-poly(ethylene oxide) diblock copolymer (PS-b PEO) as templates coupled with sol-gel chemistry. Both the functionalized Au nanoparticles (NPs) and titania precursors are selectively incorporated into the PEO domain. A series of morphologies ranging from nanodot to nanowire arrays is formed and the mechanism of morphological evolution is discussed. Such hybrid films exhibit characteristic localized surface plasmon resonance bands originating from the coupling between neighboring Au NPs spatially located in a controlled manner. PMID- 21638467 TI - Macromol. Rapid commun. 22/2009. PMID- 21638468 TI - Macromol. Rapid commun. 22/2009. PMID- 21638469 TI - Polymeric micelles induced by interpolymer complexation. AB - The self-assembly of amphiphilic diblock copolymers in block selective solvents is well documented in the scientific literature. It gives rise to micelles in which the insoluble blocks form a core, which is surrounded by a corona that contains the soluble blocks. Another method to trigger micellization consists in introducing additional non-covalent interactions such as electrostatic interactions or hydrogen bonding in an initially soluble block copolymer. The non covalent complexes that result from these interactions should be insoluble in order to induce micellization. Such insoluble complexes can be generated by mixing, in a non-selective solvent for all the individual blocks, two block copolymers or a block copolymer and a homopolymer, that contain mutually interacting blocks. This mixing process may, therefore, lead to insoluble non covalent complexes, which further aggregate into micellar cores stabilized by the uncomplexed blocks. Such a strategy has been successfully implemented in both aqueous and non-aqueous solvents to create interesting stimuli-responsive systems, mainly using ionic interactions and hydrogen bonding. This feature article will summarize these approaches. PMID- 21638470 TI - Phenylenevinylene Block Copolymers via Ring-Opening Metathesis Polymerization. AB - Fully conjugated block copolymers containing 1,4- and 1,3-phenylenevinylene repeating units can be prepared by the sequential ring opening metathesis polymerization of strained cyclophanedienes, initiated by ruthenium carbene complexes (Grubbs metathesis catalysts). The molecular weight of the constituent blocks can be tightly controlled by changing the catalyst to monomer ratio and the volume fraction of the block copolymers independently tailored by the ratio of the monomers employed. Extensive phase separation between the constituent blocks is observed in thin films of these polymers by atomic force microscopy and efficient energy transfer between blocks containing 1,4- and 1,3 phenylenevinylene units can be seen in the photoluminescence of these materials. PMID- 21638471 TI - A main-chain de vries smectic liquid crystal polymer prepared by hoveyda-grubbs catalyst initiated acyclic diene metathesis polymerization. AB - A main-chain liquid crystalline polymer has been obtained by applying a Hoveyda Grubbs 2nd generation catalyst in acyclic diene metathesis polymerization (ADMET) of a monomer containing on one end a terminal dimethylvinylsilyl group and at the other end a terminal C?C double bond. This material showed an interesting Iso-de Vries SmA* - SmC* - Glass phase transition with a very small layer shrinkage on progressing from the SmA* phase into the SmC* phase. Will this material present a helical structure along the fiber axis in the SmC* temperature range? Several physical characterization methods including XRD, optical observation, and microtome technique have been used to investigate the internal structural organization in this liquid crystalline fiber. PMID- 21638472 TI - Cyclopolymerization of Nonconjugated Dienes with a Tridentate Phenoxyamine Hafnium Complex Supported by an sp(3) -C Donor: Isotactic Enchainment and Diastereoselective cis-Ring Closure. AB - Cyclopolymerization of nonconjugated dienes produces poly(methylene-1,3 cycloalkanes) and provides a pathway to a number of stereochemically complex polymers. Activation of a diastereomeric mixture of a six-membered metallacycle complex (rac-1) in the presence of 1,5-hexadiene produced poly(methylene-1,3 cyclopentane) (PMCP) with >98% cyclization of the diene monomer. The catalyst was found to cyclopolymerize 1,5-hexadiene with relatively high activity. The microstructure of the PMCP furnished by rac-1 was found to contain a high proportion of cis-cyclopentane rings (sigma = 0.70-0.74) and a relatively high isotactic content (alpha = 0.93-0.96). These are the first cis-enriched isotactic cyclopolymers of 1,5-hexadiene. Cyclopolymerization of 1,6-heptadiene with rac 1/B(C(6) F(5) )(3) produced poly(methylene-1,3-cyclohexane) containing 97% cis isotactic rings. This is the first report of this highly isotactic and diastereomerically-pure microstructure. PMID- 21638473 TI - Large stokes-shift fluorescent silica nanoparticles with enhanced emission over free dye for single excitation multiplexing. PMID- 21638474 TI - Annular Multi-Shelled Spherulites in Interiors of Bulk-Form Poly(nonamethylene terephthalate). AB - Morphology of crystallized spherulites in poly(nonamethylene terephthalate) (PNT) in bulk forms, instead of thin-film forms, was investigated using scanning electron microscopy (SEM), polarized-optical microscopy (POM), and transmission electron microscopy (TEM). Ring textures were found to exist on the fractured surfaces or microtomed films from bulk PNT samples. By further SEM and TEM examinations, the fracture surfaces and microtomed films from the interior of the bulk display similar ring-banded patterns as those in thin films cast on glass slides. Although the cast PNT thin films in two-dimensional growth are known to display both Type-1 ring bands as majority and Type-2 ring bands as minority, this study further shows that the interior of PNT bulk exhibits mainly Type-1 (single rings with narrower spacing than Type-2) ring bands, and Type-2 spherulites exist sporadically in three-dimensional forms. From these analyses on the ring bands in the interiors of bulk-form PNT crystallized at several specific temperatures, it can be proposed that the growth of spherulites in three dimensions is layer-by-layer, packing into a multi-shells structure in three dimensional ring-banded spheres. PMID- 21638475 TI - Formation of Two Kinds of Hexagonally Arranged Structures in ABC Triblock Copolymer Thin Films Induced by a Strongly Selective Solvent Vapor. AB - An order-order transition (OOT) in the sequence of a hexagonally arranged core shell cylinder to a double-hexagonally arranged dot in polystyrene-block poly(butadiene)-block-poly(2-vinylpyridine) (SBV) triblock copolymer thin films is reported to be induced upon exposure to a solvent vapor that is strongly selective for the two end blocks. These two kinds of hexagonally arranged structures could form when the film thickness is 44, 123, and 223 nm. When the film thickness is decreased to 13 nm, the ordered structure is absent. The sizes of the core-shell cylinder structures formed with the same annealing time in films of different thickness are compared to address the effects of film thickness on the phase structure. The mechanism is analyzed from the total surface area of the blocks and the effective interaction parameter in the solvent vapor. PMID- 21638476 TI - A Facile Strategy for Preparation of alpha-Heterobifunctional Polystyrenes with Well-Defined Molecular Weight. AB - A facile strategy for synthesis of alpha-heterobifunctional polystyrenes is reported. The novel functional polystyrenes have been successfully synthesized via a combination of atom transfer radical polymerization (ATRP) and chemical modification of end-functional groups. First, epsilon-caprolactone end-capped polystyrenes with controlled molecular weight and low polydispersity were prepared by ATRP of styrene using alpha-bromo-epsilon-caprolactone (alphaBrCL) as an initiator. Then, removal of the terminal bromine atom was performed with iso propylbenzene in the presence of CuBr/PMDETA. Finally, ring-opening modifications of the caprolactone group were carried out with amines, n-butanol and H(2) O to produce novel polystyrenes containing two different functional groups at one end. PMID- 21638477 TI - Synthesis of a photoresponsive liquid-crystalline polymer containing azobenzene. AB - The synthesis of an oriented liquid-crystalline photoresponsive polymer, prepared by polymerization of mono- and di-acrylates, both of which contain azobenzene chromophores, is reported. The prepared free-standing polymer film shows strong reversible photoinduced deformation upon exposure to unpolarized UV light at 366 nm, as a result of an optically induced isomeric change of the azobenzene moieties in the polymer network. The synthesis process is relatively simple and more efficient compared to conventional ones, and can be used to synthesize other liquid-crystalline photoresponsive polymers. The use of this photoresponsive polymer film as an optical high-pass/low-pass switch under UV or natural light irradiation for a laser beam is demonstrated. This photoresponsive polymer may have applications in robotic systems, artificial muscles, and actuators in microelectromechanical systems (MEMS) and labs on chips. PMID- 21638478 TI - Polymer-Infiltrated Aligned Carbon Nanotube Fibers by in situ Polymerization. AB - Carbon nanotube-polymer composite fibers are obtained by infiltration of a monomer liquid into aligned carbon nanotube aerogel fibers with subsequent in situ polymerization. The monomer, methyl methacrylate (MMA), was infiltrated into the aerogel fibers of multi-walled carbon nanotubes (MWNTs) at room temperature and subsequently polymerized at 50 degrees C into poly(methyl methacrylate) (PMMA). Cross-sections of the PMMA/MWNT composite fibers showed that the PMMA filled the spaces of the nanotube fibers and bound the nanotubes together. PMMA in the composite fibers exhibited local order. The resultant composite fibers with 15 wt.-% nanotube loading exhibited a 16-fold and a 49-fold increase in tensile strength and Young's modulus, respectively, compared to the control PMMA. PMID- 21638479 TI - Construction of Large-Scale Highly Ordered Macroporous Monoliths of pi-Conjugated Polymers. AB - Large scale of well-ordered macroporous pi-conjugated polymer monoliths have been successfully prepared through a new approach using micrometer-sized naphthalene crystals as templates. The macroporous monoliths of poly(p-phenylenevinylene) (PPV) and poly(p-phenyleneethynylene) (PPE) grew along the unidirectional freezing direction inside the template naphthalene crystals which lead to the formation of controlling morphologies and homogeneous diameters. The polymer monoliths show straight and lamella macroporous structures. The diameters of pores and the thickness of pore walls can be controlled by tuning the freezing temperature. PMID- 21638480 TI - A magnetically tunable colloidal crystal film for reflective display. AB - A general approach to fabricate a magnetic field responsive colloidal crystal film has been developed. This is carried out by depositing monodisperse Fe(3) O(4) /PS composite magnetic nanospheres on the surface of an agarose-gel coated substrate. The optical properties of the resultant film can be easily controlled by an external magnetic field, which is caused by assembly of the magnetic nanospheres and alteration of the interparticle distance. With the help of a designed array of small magnets, both the color and pattern of the film can be conveniently modulated and the tuning range covers almost the whole visible spectrum. This work will be important for the potential application of monodisperse magnetic nanospheres in fabricating novel sensors, displays and optoelectronic devices. PMID- 21638481 TI - Macromol. Rapid commun. 23/2009. PMID- 21638482 TI - Acrylate free radical polymerization: from mechanism to polymer design. PMID- 21638483 TI - Historic account of the development in the understanding of the propagation kinetics of acrylate radical polymerizations. AB - Acrylates and methacrylate polymers are widely used in a variety of applications such as coatings, adhesives, and synthetic rubbers. In a continuous effort to increase the understanding and control over the radical (co)polymerizations of acrylate monomers the kinetic parameters, such as the propagation and termination rate constants, need to be determined. In particular, the propagation rate coefficients of the family of acrylate monomers have been a topic of intense scientific study over the last 10 years or so. In the last decade an almost full understanding of the complicated aspects of acrylate radical polymerizations has been established. It is the purpose of this essay to give a short historic account of the developments in this understanding of acrylate radical polymerizations. PMID- 21638484 TI - EPR Analysis of n-Butyl Acrylate Radical Polymerization. AB - Via electron paramagnetic resonance (EPR) spectroscopy, concentrations of secondary propagating radicals (SPRs) and tertiary mid-chain radicals (MCRs) in n butyl acrylate solution polymerization were measured. The EPR spectrum is dominated by the 4-line spectrum of SPRs at -50 degrees C and by the 7-line spectrum of MCRs at +70 degrees C. At intermediate temperatures, a third spectral component is seen, which is assigned to an MCR species with restricted rotational mobility. The MCR components are produced by 1,5-hydrogen shift (backbiting) of SPRs. The measured ratio of MCRs to SPRs allows for estimating the rate coefficient k pt for monomer addition to a mid-chain radical. For 70 degrees C, k pt is obtained to be 65.5 L . mol(-1) . s(-1) . PMID- 21638485 TI - Time-resolved electron spin resonance spectroscopy of radicals formed during free radical polymerizations of alkyl acrylates. AB - Time-resolved electron spin resonance (TR ESR) spectra allows direct observation of transient radicals (primary propagating radicals) obtained by the addition reaction of a diphenylphosphinoyl radical to acrylates. The addition rate constants (k(2) ) of the diphenylphosphinoyl radical to the acrylates (initiation rate constants) were determined by the monomer concentration dependence of the decay rate of the diphenylphosphinoyl radical. The value of k(2) was 1.9 * 10(7) s(-1) . M(-1) for the case of t-butyl acrylate, while the k(2) values of ethyl, n-butyl, 2-ethylhexyl, and dodecyl acrylates were about 1 * 10(7) s(-1) . M(-1) . A combination of steady state ESR and TR ESR provided information on the kinetics both of initiation and propagation processes in the radical polymerization of acrylates. PMID- 21638486 TI - Effect of intramolecular transfer to polymer on stationary free radical polymerization of alkyl acrylates, 4 - consideration of penultimate effect. AB - A penultimate model of acrylate polymerization has been proposed to account for reduced reactivity of radicals formed by monomer addition to midchain radicals. New expressions derived for polymerization rate, number-average degree of polymerization and branching level are used for a comparative analysis of the penultimate and terminal models. The penultimate model has been also implemented into the simulation program PREDICI to conduct a comparative analysis of time dependencies of conversion, cumulative number-degree of polymerization and cumulative branching levels calculated for the different models. The calculations show that, depending on radical reactivity ratio s(i) and monomer concentrations, the predictions from the penultimate model significantly deviate from those of the terminal model. PMID- 21638487 TI - Surface-Initiated PLP-SEC of Butyl Acrylate and Styrene from Silica Nanoparticles. AB - The pulsed-laser polymerization size-exclusion chromatography (PLP-SEC) technique has been successfully applied to the measurement of the propagation rate coefficient, k(p) , of the surface-initiated polymerizations of butyl acrylate (BA) and styrene, using the silica-immobilized bipedal initiator 4,4'-azobis(4 cyano-N-(3"-triethoxysilylpropyl)-valeric amide) (ACTA). The molecular weight distribution (MWD) of grafted poly(BA) polymerized at 25 degrees C was structureless, whereas at 5 degrees C reaction temperature, the MWD exhibited a typical PLP structure with two inflection points. In case of styrene the SEC trace obtained at 26 degrees C is well structured and shows four inflection points. The propagation rate coefficients k(p) of the surface-initiated polymerizations were about 22% higher in the case of BA and about 27-28% higher in the case of styrene compared to the IUPAC benchmark data. PMID- 21638488 TI - Transfer to Polymer and Long-Chain Branching in PLP-SEC of Acrylates. AB - Pulsed laser polymerization (PLP) combined with size exclusion chromatography (SEC) is the method of choice for determining propagation rate coefficients. The influence of the long-chain branching in PLP-SEC is investigated using multiple detection SEC and a recently developed method to detect long-chain branching [P. Castignolles, R. Grab, M. Parkinson, M. Wilhelm, M. Gaborieau, Polymer 2009, 50, 2373.] While little or no long-chain branching is detected in poly(n-butyl acrylate), the error in relevant molecular weights of poly(2-ethylhexyl acrylate) is large (30-100%) due to long-chain branching. Possible variations of propagation rate coefficient with alkyl groups in alkyl acrylates or with the solvent have to be reconsidered. PMID- 21638489 TI - Chain Transfer to Polymer and Branching in Controlled Radical Polymerizations of n-Butyl Acrylate. AB - Chain transfer to polymer (CTP) in conventional free-radical polymerizations (FRPs) and controlled radical polymerizations (ATRP, RAFT and NMP) of n-butyl acrylate (BA) has been investigated using (13) C NMR measurements of branching in the poly(n-butyl acrylate) produced. The mol-% branches are reduced significantly in the controlled radical polymerizations as compared to conventional FRPs. Several possible explanations for this observation are discussed critically and all except one refuted. The observations are explained in terms of differences in the concentration of highly reactive short-chain radicals which can be expected to undergo both intra- and inter-molecular CTP at much higher rates than long chain radicals. In conventional FRP, the distribution of radical concentrations is broad and there always is present a significant proportion of short-chain radicals, whereas in controlled radical polymerizations, the distribution is narrow with only a small proportion of short-chain radicals which diminishes as the living chains grow. Hence, irrespective of the type of control, controlled radical polymerizations give rise to lower levels of branching, when performed under otherwise similar conditions to conventional FRP. Similar observations are expected for other acrylates and monomers that undergo chain transfer to polymer during radical polymerization. PMID- 21638490 TI - Consideration of Macromonomer Reactions in n-Butyl Acrylate Free Radical Polymerization. AB - n-Butyl acrylate (BA) starved-feed solution semibatch experiments with varying final polymer content and monomer feed times were carried out at 138 degrees C. A full mechanistic model of the system implemented in Predici includes intermolecular chain transfer to polymer and macromonomer propagation as well as backbiting, chain scission, and midchain radical propagation and termination. The importance of macromonomer propagation under these conditions of industrial interest is illustrated by experiment and simulation, with the macromonomer reaction responsible for the significant increase in polymer weight-average molecular weight ($?overline M _{?rm w}$) with time. Rate coefficients for macromonomer propagation (k(mac) ) and beta-scission (k(beta) ) of k(mac) /k(p) = 0.55 and k(beta) = 12 s(-1) (with k(p) the rate coefficient for BA chain-end propagation) provide a good representation of experimental $?overline M _{?rm w}$ and macromonomer end group data at 138 degrees C. PMID- 21638491 TI - Synthesis of a macromonomer library from high-temperature acrylate polymerization. AB - The auto-initiated high temperature acrylate polymerization represents a versatile route for the synthesis of macromonomer building blocks. Various macromonomers were synthesized via this route based on methyl, ethyl, n-butyl, t butyl, 2-ethylhexyl, isobornyl and 2-[[(butylamino)carbonyl]oxy]ethyl acrylate. The synthesis requires a temperature of 140 degrees C and is carried out in a 5 wt.-% solution of hexyl acetate. The macromonomer library is fully characterized via electrospray ionization mass spectrometry (ESI-MS). The amount of macromonomers containing the geminal double bond lies in between 82 and 95%, depending on the monomer type. The achievable molecular weight of the macromonomers is located between 800 and 2 000 g.mol(-1) with a polydispersity of close to 1.6. In addition, it is demonstrated that radical initiators are useful add-ons (to circumvent the inhibition time observed during initiator-free synthesis) without interfering in the actual polymerization as no initiator fragment containing products are identified via high resolution mass spectrometry. PMID- 21638492 TI - Synthesis of cationic polyelectrolytes by inverse microemulsion polymerization. AB - The inverse microemulsion copolymerization of acrylamide and [2 (acryloyloxy)ethyl]- trimethylammonium chloride (Adamquat) was investigated using different reactors (batch, semicontinuous, and CSTR). It was found that formation of long branches through extensive intermolecular chain transfer occurred at low temperature (35 degrees C). Molecular weights and long branching levels were correlated and decreased as batch > semicontinuous > CSTR. These differences affected the performance of the copolymers as flocculants. Results indicated that successful flocculant should contain a good balance between short and long chains. PMID- 21638493 TI - Nitroxide-mediated copolymerization of 2-hydroxyethyl acrylate and 2 hydroxypropyl acrylate: copolymerization kinetics and thermoresponsive properties. AB - Poly(2-hydroxyethyl acrylate) (PHEA) is an interesting biomaterial with similar biocompatibility as the widespread used poly(2-hydroxyethyl methacrylate). Poly(2 hydroxypropyl acrylate) (PHPA) is a less frequently studied polymer that exhibits thermoresponsive behavior in water. Therefore, copolymers of HEA and HPA are interesting thermoresponsive polymers to evaluate as potential biomaterials. Here, we report the copolymerization kinetics of HEA and HPA by nitroxide mediated polymerization, employing Blocbuilder(r) initiator and SG-1 free nitroxide, revealing an ideal random copolymerization. Furthermore, the thermoresponsive properties of the resulting copolymers are discussed, whereby the cloud points are related to both polymer composition as well as polymer concentration. PMID- 21638494 TI - Star-Shaped Polyacrylates: Highly Functionalized Architectures via CuAAC Click Conjugation. AB - Well-defined functional star-shaped polymer structures with up to 29 arms have been successfully synthesized by the combination of atom transfer radical polymerization (ATRP) and click chemistry. First, azide end-functionalized poly(isobornyl acrylate) (PiBA) star-shaped polymers were prepared by successive ATRP and bromine substitution. Subsequently, alkyne end-functionalized molecules and polymers were introduced onto the star-shaped PiBA bearing pendant azide moieties by copper-catalyzed azide-alkyne cycloaddition (CuAAC). The possibilities and limits for the CuAAC on such highly branched polyacrylates are described. PMID- 21638495 TI - Macromol. Rapid commun. 24/2009. PMID- 21638496 TI - Covalently Scaffolded Inter-pi-System Orientations in pi-Conjugated Polymers and Small Molecule Models. AB - Organic pi-conjugated polymers have emerged as one of the most fascinating classes of materials as they have found utility in a host of plastic electronics technologies. The distance between pi-systems and their relative orientation dictate energy/charge transfer, conductivity, and photophysical properties of these materials in bulk. This Feature Article discusses pi-conjugated polymers and model compounds in which specific inter-pi-system interactions are covalently enforced and the effect that the scaffolding has on optoelectronic properties. PMID- 21638497 TI - Steric coordination control of interchain interactions in conducting metallopolymers. AB - The solution processability of a conducting metallopolymer (CMP1) based on a 2,2' bipyridyl (bipy) derivatized poly(p-phenylene vinylene) (PPV) backbone has been accomplished by the strategic placement of sterically demanding mesityl side chains. The enhanced solubility of CMP1 can be traced to the prevention of coordinative crosslinking between polymer chains. The sterically enforced 1:1 bipy/metal ratio was confirmed by job analysis of absorption spectroscopic titration data. In addition to enhanced processability, this strategy also leads to twice as many metal ions, and consequently twice the charge, on CMP1 versus traditional bipyridyl-PPV metallopolymers that are typified by a 2:1 bipy/metal ratio with certain metals. PMID- 21638498 TI - Transparent and conductive polyethylene oxide film by the introduction of individualized single-walled carbon nanotubes. AB - It is demonstrated that an optically transparent and electrically conductive polyethylene oxide (PEO) film is fabricated by the introduction of individualized single-walled carbon nanotubes (SWNTs). The incorporated SWNTs in the PEO film sustain their intrinsic electronic and optical properties and, in addition, the intrinsic properties of the polymer matrix are retained. The individualized SWNTs with smaller diameter provide high transmittance as well as good electrical conductivity in PEO films. PMID- 21638499 TI - Enhancement of oxidative stability of polyfluorenes for direct thermal lithography. AB - A homopolymer of 9,9'-bis[4-(2-(2-tetrahydropyranyloxy)ethoxy)phenyl]fluorene and its copolymers with 3,4-benzothiadiazole and 4,7-di(3(4-n-octylphenyl)-2-thienyl) 2,1,3-benzothiadiazole were synthesized to produce a series of thermally reactive blue, green, and red luminescent polymers. Thermolytic removal of the tetrahydropyran (THP) group from polymer films, rendered the films insoluble due to the formation of hydroxyl groups on the termini of side chains. Thermal removal of the THP was lowered by up to 200 degrees C, when acid is present in the films. These polymers were found applicable to patterning by NIR direct thermal lithography, in conjunction with a NIR dye and thermal acid generator. The presence of the phenyl groups at the 9-site carbon was found necessary to eliminate fluorenone formation, and enhance the colour purity of the material. PMID- 21638500 TI - Synthesis and Fluorescent Properties of Biodegradable Hyperbranched Poly(amido amine)s. AB - Disulfide-functionalized hyperbranched poly(amido amine)s (HPAMAMs) were synthesized by Michael addition polymerization of N,N'-cystaminebisacrylamide and 1-(2-aminoethyl)piperazine. The novel HPAMAMs displayed bright fluorescence, and the emissions bands cover nearly the whole visible wavelength range. When polymer solutions were excited at 330-385, 460-490, and 510-550 nm, blue, green, and red solutions were observed, respectively. The HPAMAMs are biodegradable and they can be easily cleaved by 2-mercaptoethanol or glutathione, leading to a decrease in the fluorescence intensity. Studies of applications of the biocompatible and biodegradable HPAMAMs in fluorescence imaging technology and biological science are in progress. PMID- 21638501 TI - Melt dispersion and electrospinning of non-functionalized multiwalled carbon nanotubes in thermoplastic polyurethane. AB - Nanoscale fibers with embedded, aligned, and percolated non-functionalized multiwalled carbon nanotubes (MWCNTs) were fabricated through electrospinning dispersions based on melt-compounded thermoplastic polyurethane/MWCNT nanocomposite, with up to 10 wt.-% MWCNTs. Transmission electron microscopy indicated that the nanotubes were highly oriented and percolated throughout the fibers, even at high MWCNT concentrations. The coupling of efficient melt compounding with electrospinning eliminated the need for intensive surface functionalization or sonication of the MWCNTs, and the high aspect ratio as well as the electrical and mechanical properties of the nanotubes were retained. This method provides a more efficient technique to generate one-dimensional nanofibers with aligned MWCNTs. PMID- 21638502 TI - Synthesis of oligothiophene-layered polymers. AB - Polymers with layered pi-electron systems comprising bithiophenes and quaterthiophenes based on a xanthene skeleton have been synthesized by an iron catalyzed oxidative coupling reaction. The polymers are well soluble in common organic solvents, and polymer thin films are readily obtained by a spin-coating or casting technique. The synthesized polymers exhibit good thermal stability, and their 10% weight loss temperatures are approximately 400 degrees C. These behaviors of the polymers suggest that they can be used as opto-electronic devices such as hole-transporting materials. PMID- 21638503 TI - Synthesis of POSS-Functionalized Polyisobutylene via Direct Initiation. AB - POSS-functionalized polyisobutylenes (PIBs) were synthesized by carbocationic polymerization using an epoxy-POSS/TiCl(4) initiating system in hexane/methyl chloride (60:40 v/v) solvent mixture at -80 degrees C. (1) H NMR spectroscopy verified the incorporation of one epoxy-POSS per polymer chain. Light scattering and TEM analysis demonstrated the formation of 50-100 nm sized aggregates and micron-sized clusters. PMID- 21638504 TI - A facile route to prepare organic/inorganic hybrid nanomaterials by 'click chemistry'. AB - An organosilane with an alkyne group at the non-condensable end, [(2 propynylcarbamate)propyl]triethoxysilane, has been synthesized. Condensation of this organosilane with tetraethoxysilane can be achieved by a co-condensation strategy to produce silica nanoparticles with surface alkyne functionality. The size and uniformity of size distribution of the silica nanoparticles are influenced by varying the concentration of the added organosilane. The alkyne functionalized silica nanoparticles are coupled directly with azide-modified polymers by 'click chemistry' to yield organic-inorganic hybrid nanomaterials. PMID- 21638505 TI - The Self-Assembled Structure of the Diblock Copolymer PCL-b-P4VP Transforms Upon Competitive Interactions with Octaphenol Polyhedral Oligomeric Silsesquioxane. AB - This paper describes the miscibility and self-assembly, mediated by hydrogen bonding interactions, of new block copolymer/nanoparticle blends. The morphologies adopted by the immiscible poly[(epsilon-caprolactone)-block-(4-vinyl pyridine)] (PCL-b-P4VP) diblock copolymer changes upon increasing the number of competitive hydrogen-bonding interactions after adding increasing amounts of octaphenol polyhedral oligomeric silsesquioxane (OP-POSS). Transmission electron microscopy reveals morphologies that exhibit high degrees of long-range order, such as cylindrical and spherical structures, at relatively low OP-POSS contents, and short-range order or disordered structures at higher OP-POSS contents. Analyses performed using differential scanning calorimetry, wide-angle X-ray diffraction, and FT-IR spectroscopy provide positive evidence that the pyridyl units of the P4VP block are significantly stronger hydrogen-bond acceptors toward the OH group of OP-POSS than are the C?O groups of the PCL block, thereby resulting in excluded and confined PCL phases. PMID- 21638506 TI - Highly Effective and Green Catalytic Approach Toward alpha,omega-Dihydroxy Telechelic Poly(trimethylenecarbonate). AB - alpha,omega-Dihydroxy-telechelic poly(trimethylenecarbonate), HO-PTMC-OH, is synthesized from the controlled "immortal" ring-opening polymerization (ROP) of trimethylene carbonate under mild conditions (bulk, 60 degrees C), using ZnEt(2) or, more efficiently, [(BDI)Zn(N(SiMe(3) )(2) )] (BDI = CH(CMeNC(6) H(3) -2,6 iPr(2) )(2) ) as catalyst precursor, in the presence of a diol HO-R-OH (R = (CH(2) )(2) or CH(2) C(6) H(4) CH(2) ; 0.5-10 equiv. vs Zn) acting both as co initiator and chain transfer agent. Alternatively, HO-PTMC-OH is prepared upon hydrogenolysis of HO-PTMC-OCH(2) Ph, initially prepared from the ROP of TMC using the [(BDI)Zn(N(SiMe(3) )(2) )]/PhCH(2) OH system, under smooth operating conditions using Pd/charcoal. Well-defined dihydroxy-functionalized PTMCs of molar mass ranging from $?overline M _{?rm n}$ = 2 000 to 109 500 g . mol(-1) were thus quantitatively obtained and fully characterized by NMR, MALDI-TOF-MS and SEC analyses. The versatility of this "immortal" ROP allows the preparation of alike alpha,omega-functional polyester such as linear HO-poly(lactide)-OH, as well as star polymers such as the glycerol-based PTM-OH(3) . PMID- 21638507 TI - Non-Fouling Character of Poly[2-(methacryloyloxy)ethyl Phosphorylcholine] Modified Gold Surfaces Fabricated by the 'Grafting to' Method: Comparison of its Protein Resistance with Poly(ethylene glycol)-Modified Gold Surfaces. AB - Poly[2-(methacryloyloxy)ethyl phosphorylcholine] -modified gold surfaces, which have been newly prepared by a 'grafting to' method using a series of monosulfanyl terminated PMPC, are characterized by protein adsorption experiments based on surface plasmon resonance spectroscopy and ellipsometry measurements. The extent of BSA adsorption on PMPC-modified surfaces was systematically reduced for thicker PMPC layers, thus the number of MPC units on the gold surface appears to be an important factor for the excellent protein resistance offered by PMPC modified gold surfaces fabricated by the 'grafting to' method, which is sharp contrast to that of PEG tethered chains. PMID- 21638508 TI - Highly Asymmetric Phase Diagram of a Poly(1,2-octylene oxide)-Poly(ethylene oxide) Diblock Copolymer System Comprising a Brush-Like Poly(1,2-octylene oxide) Block. AB - The phase diagram of a series of poly(1,2-octylene oxide)-poly(ethylene oxide) (POO-PEO) diblock copolymers is determined by small-angle X-ray scattering. The Flory-Huggins interaction parameter was measured by small-angle neutron scattering. The phase diagram is highly asymmetric due to large conformational asymmetry that results from the hexyl side chains in the POO block. Non-lamellar phases (hexagonal and gyroid) are observed near f(PEO) = 0.5, and the lamellar phase is observed for f(PEO) >= 0.5. PMID- 21638509 TI - Prostasomal DNA characterization and transfer into human sperm. AB - Human prostasomes, exosome-like microvesicles secreted by acinar cells of the prostate gland, contain chromosomal DNA. Agarose gel electrophoresis of DNA from seminal prostasomes displayed fragments of over 12 kb and smaller, with a distinct band around 1 kb that was excised, cloned, and sequenced. The sequences showed 8 out of 25 clones (32%) originating from genes. We elaborated the concept further by carrying out a genome-wide DNA copy number analysis of prostasomal DNA, hypothesizing that human prostasomes contain fragments of DNA randomly selected from the entire genome. Acridine orange-stained prostasomes were incubated with freshly prepared sperm for different times, and a transfer of acridine orange-stained prostasomal DNA to sperm (preferentially the head region) was observed. Fluorescence microscopy of slices in the center of 14 optical slides of the sperm head displayed an even fluorescence rather than a halo-like one, indicating DNA-uptake rather than just binding along the sperm head membrane. PMID- 21638510 TI - Sperm storage in the female reproductive tract of Scotophilus heathii: role of androgen. AB - The aim of the present study was to investigate the mechanism of androgen mediated, prolonged sperm-storage in the female reproductive tract of the bat, Scotophilus heathii. The bat treated in vivo with flutamide, an androgen antagonist, showed loss of spermatozoa at the storage site, utero-tubal junction. Immunocytochemistry and Western blot analysis revealed the presence of increased expression of Bcl2 in the epithelial cell lining of the utero-tubal junction during the period of sperm-storage. Treatment with testosterone in vitro caused a significant dose-dependent increase in expression of the survival factor Bcl2, whereas treatment with flutamide together with testosterone caused a significant decline in Bcl2 in the utero-tubal junction of S. heathii. Together with the expression of Bcl2, the utero-tubal junction also expresses the death signal, caspase3. Expression of caspase3 decreased during January, but increased in February during the late stage of sperm storage. Androgen stimulated Bcl2 synthesis in the utero-tubal junction via the non-genomic MAP kinase signaling pathway. In conclusion, this study suggests that androgen promotes sperm storage in S. heathii by stimulating the survival factor Bcl2 in the utero-tubal junction. It is further hypothesized that a balance between the survival factor, Bcl2, and the death signal, caspase3, determines the duration of sperm storage in S. heathii. PMID- 21638511 TI - Setting up asymmetric cell division in the mouse egg. PMID- 21638512 TI - Cortisol and testosterone in Filipino young adult men: evidence for co-regulation of both hormones by fatherhood and relationship status. AB - OBJECTIVES: Although cortisol (CORT) may suppress testosterone (T) production under stress, in many species males' T and CORT are co-elevated during mate acquisition or conspecific competition. It is presently unknown how CORT co varies with T in relation to fatherhood/relationship status in men. Here we evaluate associations between waking (AM) and pre-bed (PM) salivary CORT and T, and with plasma total T and luteinizing hormone. We also test whether co elevationor co-downregulation of CORT and T are present in men who are mating oriented (non-pairbonded, non-fathers) and parenting-oriented (pairbonded and/or fathers), respectively. METHODS: Data come from 630 of young adult Filipino males (21-23 years) enrolled in the Cebu Longitudinal Health and Nutrition Survey, a population-based birth cohort study in Cebu City, Philippines. RESULTS: T and CORT were positively related in AM (r = 0.37) and PM (r = 0.30) saliva samples (both P < 0.001). The positive relationship between AM measures was strengthened as caloric intake improved (interaction P < 0.05). Mating-oriented men were more likely to have co-elevated PM CORT and T (P < 0.05), defined as being in the highest tertile for both hormones, while parenting-oriented men were more likely to have co-downregulated (lowest tertile for both hormones) AM (P < 0.05) and PM (P < 0.001) CORT and T. CONCLUSIONS: CORT and T are positively related upon waking and before bed and are more likely to be co-elevated in mating-oriented men and co-downregulated in parenting-oriented men. Our findings support the interpretation that CORT and T serve complementary roles in facilitating men's mating effort. PMID- 21638513 TI - Outcomes after thyroidectomy and parathyroidectomy. AB - BACKGROUND: Previous reports on postoperative outcomes following thyroid and parathyroid surgery are limited by relatively small sample size. We report 30-day outcomes following thyroid and parathyroid surgery and analyze factors affecting length of stay (LOS) and postoperative adverse events (AEs). METHODS: The multicenter, prospective, National Surgical Quality Improvement Program (NSQIP) datasets (2007/2008) were used. Multivariable logistic regression and analysis of covariance (ANCOVA) were performed. RESULTS: Patients undergoing thyroidectomy, parathyroidectomy, or both were identified (n = 13,380, 6154, 1535, respectively). Thirty-day mortality was 0.08%, 0.16%, and 0.2%, respectively; 30 day morbidity was 3.50%, 3.02%, and 4.04%, respectively. Mean LOS values were 1.1 +/- 1.4, 1.1 +/- 2.1, and 1.4 +/- 3.1 days, respectively. Congestive heart failure, dependent functional status, dialysis dependence, and chronic corticosteroid use were significantly associated with increased LOS and postoperative AE. CONCLUSIONS: Morbidity and mortality rates following thyroid and parathyroid surgery are low. These data could be used by third-party interests, and surgeons should be aware of them to ensure their outcomes are in the national norm. PMID- 21638515 TI - Knockdown of RARB2 identifies a dual role in cancer. AB - Two chemoprevention trials have shown that retinoic acid (RA) may be harmful in patients at risk for lung cancer, and RA administration to this high-risk group results in RARB2 reactivation. Although RARB2 is thought to possess tumor suppressive activity, its expression has recently been correlated with poorer prognosis in patients with nonsmall cell lung cancer. We hypothesized that RARB2 expression is necessary for the growth and maintenance of the oncogenic phenotype in lung cancer cells in which RARB2 has not been inactivated. We tested various antisense oligodeoxynucleotides (ASO) against RARB2 in multiple lung cancer cell lines and used microarray technology to compare the patterns of gene expression following ASO treatment versus RA treatment in the A-549 lung cancer cell line. We show that ASO treatment reduces proliferation and causes apoptosis in 3 RARB2 expressing lung cancer cell lines but has no apparent effect in at least two other lung cancer cells lines having lost RARB2 expression or one normal lung RARB2-expressing cell line; we demonstrate a correlation between resulting RARB2 expression levels and cell growth; and identify transcriptional effects related to both RA and RARB2 signaling. In particular, five genes known to contribute to carcinogenesis or chemotherapeutic resistance are down-regulated following ASO treatment: three of these are up-regulated following RA treatment. This work demonstrates a dual role for RARB2 (tumor suppression and tumor promotion) and identifies a challenge with respect to using RARB2 as a target for treatment or prevention strategies. PMID- 21638516 TI - Molecular characterization of 9p21 deletions shows a minimal common deleted region removing CDKN2A exon 1 and CDKN2B exon 2 in diffuse large B-cell lymphomas. AB - Diffuse large B cell lymphoma represent the most frequent subtype of non Hodgkin's lymphoma, accounting for 30-40% of cases. Several studies have shown that CDKN2A and CDKN2B deletions are frequent in these lymphomas. These genes encode the P14ARF, CDKN2A, and CDKN2B proteins which play a key role in the control of the G1/S transition of the cell cycle. Using array CGH and a quantitative multiplex PCR method, we have previously identified such deletions in 36% of cases at diagnosis. Using a walking strategy to approach the breakpoints of these deletions we could identify the breakpoints junction of thirteen deletions in 11 patients and of two unbalanced translocation leading to a loss of these genes. A minimal common deleted region of 22.4 kb containing exon 1beta of CDKN2A encoding P14ARF and exon 2 of CDKN2B encoding CDKN2B was present in all cases but one. Analysis by quantitative RT PCR showed that the expression level of these genes was decreased in these patients as compared with non deleted cases and that this level was correlated with the deletion status of P14ARF, CDKN2A, and CDKN2B. Analysis of the breakpoint sequences showed that some of them were clustered within a few hundred base-pairs suggesting, even if we failed to identify any clear recombination prone sequences, that in these deletions the rearrangement results from non-random mechanisms. PMID- 21638517 TI - Multiplex ligation-dependent probe amplification and fluorescence in situ hybridization to detect chromosomal abnormalities in chronic lymphocytic leukemia: a comparative study. AB - Chronic lymphocytic leukemia (CLL) is a clinically heterogeneous disease characterized by recurrent chromosomal aberrations of prognostic significance. We aimed to evaluate the potential of the multiplex ligation-dependent probe amplification (MLPA) assay to detect genomic alterations in CLL. Highly purified (>90%) peripheral mononuclear CD19+ cell populations from 100 untreated CLL patients (pts) in early stage disease (Binet stage A) were included in this study. All samples were investigated by fluorescence in situ hybridization (FISH) for the presence of trisomy 12 and 17p13.1, 11q22.3, and 13q14.3 deletions. For MPLA analysis, DNA was amplified by means of two commercially available probes sets allowing the simultaneous screening of 56 genomic sequences. Overall, a high degree of concordance (95%) between MPLA and FISH results was found, if the abnormal clone was present in more than 30% of the leukemic cell population. The use of multiple MPLA probes allowed the fine-mapping of the 13q14 deletion and the identification of intragenic or small alterations undetected by FISH. Moreover, additional alterations in 2p24 (MYCN) (3 pts), 8q24 (MYC) (1 pt), 9p21 (CDKN2A2B) (1 pt), 1q21 (LMNA) (1 pt), and 6q25-26 (1 pt) regions not covered by a standard FISH assay were detected and all confirmed by FISH. Our data extend previously limited evidence that MLPA may represent a useful technique for the characterization of well-known lesions as well as the investigation of additional genomic changes in CLL. PMID- 21638518 TI - The DNA methylome of benign and malignant parathyroid tumors. AB - The role of DNA methylation of CpG islands in parathyroid tumorigenesis has not been analyzed in an unbiased, systematic fashion. DNA was isolated from normal and pathologic parathyroid tissues, bisulphite modified and analyzed using the Infinium HumanMethylation27 BeadChip. Distinct hierarchical clustering of genes with altered DNA methylation profiles in normal and pathologic parathyroid tissue was evident. Comparing normal parathyroid tissue with parathyroid adenomas, 367 genes were significantly altered, while 175 genes significantly differed when comparing parathyroid carcinomas and normal parathyroid tissues. A comparison between parathyroid adenomas and parathyroid carcinomas identified 263 genes with significantly distinct methylation levels. Results were confirmed for certain genes in a validation cohort of 40 parathyroid adenomas by methylation-specific PCR. Genes of known or putative importance in the development of parathyroid tumors showed significant and frequent hypermethylation. DNA hypermethylation of CDKN2B, CDKN2A, WT1, SFRP1, SFRP2, and SFRP4 was associated with reduced gene expression in both benign and malignant parathyroid tumors. Treatment with 5-aza 2'-deoxycytidine of primary cell cultures restores expression of hypermethylated genes in benign and malignant parathyroid tumors. In conclusion, the unbiased, genome-wide study of the parathyroid tumor DNA methylome identified a number of genes with altered DNA methylation patterns of putative importance to benign and malignant parathyroid tumorigenesis. PMID- 21638519 TI - CBFB and MYH11 in inv(16)(p13q22) of acute myeloid leukemia displaying close spatial proximity in interphase nuclei of human hematopoietic stem cells. AB - To gain a better understanding of the mechanism of chromosomal translocations in cancer, we investigated the spatial proximity between CBFB and MYH11 genes involved in inv(16)(p13q22) found in patients with acute myeloid leukemia. Previous studies have demonstrated a role for spatial genome organization in the formation of tumorigenic abnormalities. The nonrandom localization of chromosomes and, more specifically, of genes appears to play a role in the mechanism of chromosomal translocations. Here, two-color fluorescence in situ hybridization and confocal microscopy were used to measure the interphase distance between CBFB and MYH11 in hematopoietic stem cells (HSCs), where inv(16)(p13q22) is believed to occur, leading to leukemia development. The measured distances in HSCs were compared with mesenchymal stem cells, peripheral blood lymphocytes, and fibroblasts, as spatial genome organization is determined to be cell-type specific. Results indicate that CBFB and MYH11 are significantly closer in HSCs compared with all other cell types examined. Furthermore, the CBFB-MYH11 distance is significantly reduced compared with CBFB and a control locus in HSCs, although separation between CBFB and the control is ~70% of that between CBFB and MYH11 on metaphase chromosomes. HSCs were also treated with fragile site-inducing chemicals because both the genes contain translocation breakpoints within these regions. However, treatment with fragile site-inducing chemicals did not significantly affect the interphase distance. Consistent with previous studies, our results suggest that gene proximity may play a role in the formation of cancer-causing rearrangements, providing insight into the mechanism of chromosomal abnormalities in human tumors. PMID- 21638520 TI - Safety surveillance of longitudinal databases: methodological considerations. PMID- 21638521 TI - Completely miscible nanocomposites. PMID- 21638522 TI - Dibridgehead diphosphines that turn themselves inside out. PMID- 21638523 TI - Bronsted acid catalysis: hydrogen bonding versus ion pairing in imine activation. PMID- 21638524 TI - Efficient entry to the hasubanan alkaloids: first enantioselective total syntheses of (-)-hasubanonine, (-)-runanine, (-)-delavayine, and (+)-periglaucine B. PMID- 21638525 TI - Palladium-catalyzed beta-selective direct arylation of porphyrins. PMID- 21638531 TI - A guided discovery approach for learning metabolic pathways. AB - Learning the wealth of information in metabolic pathways is both challenging and overwhelming for students. A step-by-step guided discovery approach to the learning of the chemical steps in gluconeogenesis and the citric acid cycle is described. This approach starts from concepts the student already knows, develops these further in a logical manner using easy-to-remember memory devices, and significantly reduces student learning demand. This approach incorporates the use of PowerPoint, web access, and a digital whiteboard to enhance student learning and retention. PMID- 21638532 TI - Theme-based tests: Teaching in context. AB - Theme-based tests provide an assessment tool that instructs as well by providing a single general context for a broad set of biochemical concepts. A single story line connects the questions on the tests and models applications of scientific principles and biochemical knowledge in an extended scenario. Theme-based tests are based on a set of interrelated journal articles or a fictional story derived from popular culture or local events that interweaves biochemical principles into a story line. Theme-based tests engage students during an exam and promote their creativity. PMID- 21638533 TI - A new literary metaphor for the genome or proteome. AB - Previously, the idea of a blueprint has been used to explain the genome. The concept of a play's cast of characters, the Dramatis Personae, is a more fluid metaphor that allows for mutations and time-dependent phenomena to be taken into account. It also provides an educational and mnemonic exercise for students. PMID- 21638534 TI - Screening a library of household substances for inhibitors of phosphatases: An introduction to high-throughput screening. AB - Library screening methods are commonly used in industry and research. This article describes an experiment that screens a library of household substances for properties that would make a good "drug," including enzyme inhibition, neutral pH, and nondenaturing to proteins, using wheat germ acid phosphatase as the target protein. An adaptation of the experiment appropriate for lower level biochemistry or outreach is also described. This work was supported by Wabash College through the Haines Fund for the Study of Biochemistry and the National Science Foundation through Grant DUE 0126242. PMID- 21638535 TI - Quantifying the activation of the small molecular weight G-protein ras. AB - Low molecular weight guanine-nucleotide binding proteins are molecular switches involved in a wide range of cellular functions in eukaryotes. Ras is activated when a membrane receptor is bound by a ligand and the GDP bound to the Ras protein is exchanged for GTP. Once GTP is bound, the Ras three-dimensional conformation changes and binds the kinase Raf. This protein-protein interaction can be used to "pull-down" activated Ras from a cell lysate. The amount of activated Ras is quantitated through immunoblotting and densitometry. This procedure is excellent as a multiweek investigational laboratory. PMID- 21638536 TI - Quantification of DNA by agarose gel electrophoresis and analysis of the topoisomers of plasmid and M13 DNA following treatment with a restriction endonuclease or DNA topoisomerase I. AB - A two-session laboratory exercise for advanced undergraduate students in biochemistry and molecular biology is described. The first session introduces students to DNA quantification by ultraviolet absorbance and agarose gel electrophoresis followed by ethidium bromide staining. The second session involves treatment of various topological forms of DNA with a restriction endonuclease and with a DNA topoisomerase. This session introduces students to the concept of DNA topoisomers, to the properties of different forms of DNA, and to the activity of restriction endonucleases and topoisomerases toward these forms. The exercise also involves measuring the size of linear duplex fragments of DNA by comparison of mobility with a ladder of double stranded DNA of known sizes. PMID- 21638537 TI - A complete approach for recombinant protein expression training: From gene cloning to assessment of protein functionality*. AB - A practical course was given to undergraduate biology students enrolled in the elective course "Introduction to Genetic Engineering" at the Federal University of Sao Carlos (UFSCar), Sao Paulo, Brazil. The goal of the course was to teach current molecular biology tools applied to a real research situation that could be reported by the students themselves. The purpose was to produce a plant recombinant protein and demonstrate a heretofore unreported biological activity. Cystatins, natural inhibitors of cysteine proteases, were proposed for these studies. Initially, the students searched for plant cystatin cDNA sequences in the NCBI databases and selected the Oryzacystatin I gene (ocI) from rice, Oriza sativa, as the target gene for this study. Total RNA was extracted from rice germinating seeds and primers containing restriction sites for NdeI and EcoRI were designed based on the ocI cDNA sequence and then used to amplify the open reading frame (ORF). RT-PCR amplification provided a band of the expected size for ocI ORF (309 bp). The PCR product was cut with NdeI and EcoRI restriction enzymes and cloned directly in the pET28a expression vector digested with the same enzymes. A pET28-ocI recombinant clone was selected, checked by sequencing, and used to transform Escherichia coli BL21 (DE3) expression strain. After induction of the bacteria with isopropylthiogalactoside and cellular disruption, the His-tagged OCI protein, present mainly in the soluble fraction, was purified by affinity chromatography in a nickel column. The purified protein was successfully used to inhibit fungal growth (Trichoderma reesei). The results were discussed extensively and the students contributed to the writing of this article, of which they are co-authors. PMID- 21638538 TI - Development of a green fluorescent protein-based laboratory curriculum. AB - A laboratory curriculum has been designed for an undergraduate biochemistry course that focuses on the investigation of the green fluorescent protein (GFP). The sequence of procedures extends from analysis of the DNA sequence through PCR amplification, recombinant plasmid DNA synthesis, bacterial transformation, expression, isolation, and characterization of the protein by SDS-PAGE. A survey of participants found that the majority of them were performing most of these procedures for the first time and that participants found the exercises enjoyable and considered them a significant aid to their understanding of biochemistry, cell biology, and molecular genetics. PMID- 21638539 TI - Use of visible light-based photodynamic therapy to bacterial photoinactivation. AB - The main focus of this laboratory exercise was to investigate the photodynamic therapy (PDT) acting over Streptococcus mutans. A handheld photopolymerizer and a classical photosensitizer (Rose Bengal) were used to induce photodynamic response. In this way, a suspension of S. mutans was treated with different concentrations of Rose Bengal (0-10 MUmol/liter), irradiated with a light (400 600 nm) for 20 s, and then cell viability was evaluated. It was observed that the light (per se) is not toxic, and in the dark, Rose Bengal is toxic only to the cells tested at concentrations above 5.0 MUmol/liter. Under light exposure, concentrations of Rose Bengal above 0.5 MUmol/liter killed all S. mutans. Therefore, for the purpose of our work, the photoactivation of Rose Bengal using the handheld photopolymerizer was efficient in bacteria inactivation. PMID- 21638540 TI - Determination of the Rh factor: A practical illustrating the use of the polymerase chain reaction. AB - A practical experiment on the PCR is described that has been used over several years as part of an undergraduate biochemistry and molecular biology course for chemistry students. In the first experimental session, students prepare their own DNA samples from epithelial cells of the mouth and use them as templates in the PCR. In the second session, they analyze the amplified DNA by electrophoresis and determine their Rh factor. PMID- 21638541 TI - What is worth knowing? Teaching? Learning? Understanding? PMID- 21638542 TI - The biological activity of the large-T protein of SV40 virus*. AB - Terms to be familiar with before you start to solve the test: SV40 virus, malignant transformation, in vitro DNA synthesis, template, primer, DNA polymerase, radioactive labeling, autoradiography, dideoxynucleoside triphosphates, heat denaturation, nondenaturing PAGE. PMID- 21638543 TI - Bioinformatics training programs are hot but the labor market is not*. AB - Based on a survey of academic training programs and an analysis of advertised job openings, we conclude that the labor market in bioinformatics has changed dramatically from the 1990s to the early 2000s. The number of academic training programs, as well as enrollment in these programs, expanded rapidly during this period. The expansion has created a sizeable pipeline of students who will matriculate from these programs in the near future. Yet, at the same time that this expansion in training programs occurred, demand in the bioinformatics market declined and its origins have shifted largely from industry to academe. Unless conditions in industry change dramatically in the next few years, it is likely that trainees from these programs will have difficulty finding the expected jobs in industry. PMID- 21638544 TI - Commentary: E-mail works for me. PMID- 21638545 TI - Software for teaching structure-hydrophobicity relationships. AB - We have developed a lecture and laboratory curriculum for introducing beginning undergraduate Biology students to chemical structure-function relationships. The laboratory portion of this curriculum employs cheminformatics software that provides instant feedback to help students develop an understanding of the relationship between structure and hydrophobicity. To evaluate the effectiveness of this curriculum, we measured students' understanding using an open-ended problem-based survey. Student responses to this survey improved significantly following the activities we describe, suggesting that they are effective teaching tools. This curriculum also provides a foundation for students' future structure function studies in chemistry, biochemistry, and molecular biology. PMID- 21638546 TI - Websites of note. PMID- 21638547 TI - Commentary: Metabolism: Dull, lifeless, and boring? PMID- 21638549 TI - Obituary: professor peter N. Campbell (1921-2005). PMID- 21638550 TI - Bioinformatics: Current practice and future challenges for life science education. AB - It is widely predicted that the application of high-throughput technologies to the quantification and identification of biological molecules will cause a paradigm shift in the life sciences. However, if the biosciences are to evolve from a predominantly descriptive discipline to an information science, practitioners will require enhanced skills in mathematics, computing, and statistical analysis. Universities have responded to the widely perceived skills gap primarily by developing masters programs in bioinformatics, resulting in a rapid expansion in the provision of postgraduate bioinformatics education. There is, however, a clear need to improve the quantitative and analytical skills of life science undergraduates. This article reviews the response of academia in the United Kingdom and proposes the learning outcomes that graduates should achieve to cope with the new biology. While the analysis discussed here uses the development of bioinformatics education in the United Kingdom as an illustrative example, it is hoped that the issues raised will resonate with all those involved in curriculum development in the life sciences. PMID- 21638551 TI - Development and evaluation of a peer-tutoring program for graduate students*. AB - Many interdisciplinary Ph.D. programs admit students of different educational backgrounds who receive a first year of a general curriculum education. However, student preparation for this curriculum varies, and methods are needed to provide academic support. Graduate student peer tutoring was piloted as an initiative funded by a National Institutes of Health (NIH) Initiative for Minority Student Development award to the University of Medicine and Dentistry of New Jersey Robert Wood Johnson Medical School (UMDNJ-RWJMS) and is now offered to all students in the interdisciplinary Molecular Biosciences Ph.D. program between Rutgers, The State University of New Jersey, and UMDNJ-RWJMS. Tutoring occurs individually or in small groups and has grown over the past 5 years in the number of students tutored and hours of tutoring. The program was evaluated by surveying and interviewing both tutors and students concerning process variables (e.g. awareness, frequency) and impact variables (e.g. perceived benefits, motivators), as well as by assessing changes in exam scores for the four core courses of the first-year graduate curriculum. PMID- 21638552 TI - The metabolic effects of low-carbohydrate diets and incorporation into a biochemistry course. AB - One of the challenges in teaching biochemistry is facilitating students' interest in and mastery of metabolism. The many pathways and modes of regulation can be overwhelming for students to learn and difficult for professors to teach in an engaging manner. We have found it useful to take advantage of prevailing interest in popular yet controversial weight-loss methods, particularly low-carbohydrate diets. The metabolic rationale behind these eating plans can be linked to glycolysis, the citric acid cycle, lipolysis, gluconeogenesis, ketosis, glycogen metabolism, fatty acid oxidation, and hormonal regulation. When this approach was used in undergraduate biochemistry classes at the State University of New York at Geneseo, students were highly motivated to learn the biochemical principles behind these diets. The following provides information about low-carbohydrate diet plans that will enable professors to speak authoritatively on the subject. History and studies regarding efficacy as well as biochemical metabolic effects are included. PMID- 21638553 TI - From problem-based learning to interrupted lecture: Using case-based teaching in different class formats. AB - Although many examples of using problem-based learning and case studies in teaching science occur in the literature, instructors may still have questions about when, why, and how to use these strategies, especially in different class settings. In this article, I describe my experience in using these pedagogical methods in three different class formats: a small biochemistry survey course, which I taught entirely as problem-based learning; a small organic chemistry class in which I used some case-based teaching with students working collaboratively in groups; and a large lecture-style biochemistry survey course in which I used cases as homework and in-class exercises. In each instance I describe the apparent benefits and drawbacks of each approach, drawing on the literature of how these methods may best promote learning. As part of this discussion I pose questions for instructors to help them decide which form of case-based teaching best fits with their particular teaching goals and class contexts. PMID- 21638554 TI - Physical models enhance molecular three-dimensional literacy in an introductory biochemistry course*. AB - This article reports the results of a recent study to evaluate the usefulness of physical models of molecular structures as a new tool with which to teach concepts of molecular structure and function. Of seven different learning tools used by students in this introductory biochemistry class, the use of the physical models in a laboratory was rated as most useful. These results suggest that physical models can play an important role in capturing the interest of students in the subject of molecular structure and function. These physical models often stimulate more sophisticated questions in the minds of students, which can then be more appropriately explored using computer visualization tools. PMID- 21638555 TI - Poem: Finding amino. PMID- 21638556 TI - Discovering an accessible enzyme: Salivary alpha-amylase : Prima digestio fit in ore: A didactic approach for high school students. AB - Human salivary alpha-amylase is used in this experimental approach to introduce biology high school students to the concept of enzyme activity in a dynamic way. Through a series of five easy, rapid, and inexpensive laboratory experiments students learn what the activity of an enzyme consists of: first in a qualitative then in a semi-quantitative way. They also learn how some environmental effectors can influence it. The choice of a "human body" enzyme and not an anonymous commercial enzyme is a very attractive one for students. This laboratory approach will be integrated with theoretical knowledge about alpha-amylase, starch, their physiological meaning, and biotechnological applications. PMID- 21638557 TI - An accessible two-dimensional solution nuclear magnetic resonance experiment on human ubiquitin*. AB - Solution-state nuclear magnetic resonance (NMR) is an invaluable tool in structural and molecular biology research, but may be underutilized in undergraduate laboratories because instrumentation for performing structural studies of macromolecules in aqueous solutions is not yet widely available for use in undergraduate laboratories. We have implemented an experiment that is ideal for more commonly available 4.8-7.0 Tesla, double-channel NMR instruments that would not usually be used for biomolecular NMR work. We analyzed a commercially available, (15) N-enriched human ubiquitin sample with a two dimensional correlation experiment using indirect (1) H evolution and direct (15) N detection, which produced spectra with high resolution on a spectrometer operating at 7.0 Tesla (300 MHz (1) H resonance frequency). The simplicity of the experiment makes it possible to be configured by undergraduate students with minimal supervision from the instructor. Students gain experience in acquiring multidimensional biomolecular NMR experiments, confirm that ubiquitin is stably folded, and observe the correspondence between specific signals and individual amino acids in ubiquitin. PMID- 21638558 TI - A guided inquiry experiment for the measurement of activation energies in the biophysical chemistry laboratory: Decarboxylation of pyrrole-2-carboxylate. AB - A laboratory experiment for undergraduate biophysical chemistry is described, in which the acid concentration and temperature dependences of the decarboxylation of pyrrole-2-carboxylate are measured using a continuous ultraviolet (UV) spectrophotometric assay. Data collection and analysis are structured using principles of guided inquiry. Data leading to the calculation of multiple rate constants at varying temperatures and acid concentrations can be collected within one laboratory period, using inexpensive reagents and standard instrumentation. These experiments permit determination of activation energies that are lower at high acid concentration, indicative of a subtle change in the reaction mechanism with decreasing pH. The reaction is readily observable by students as they collect UV spectrophotometry data, and the decarboxylation reaction is related to biologically relevant enzymatic reactions. PMID- 21638559 TI - Teaching about citric acid cycle using plant mitochondrial preparations: Some assays for use in laboratory courses*. AB - Potato tubers and turnip roots were used to prepare purified mitochondria for laboratory practical work in the teaching of the citric acid cycle (TCA cycle). Plant mitochondria are particularly advantageous over the animal fractions to demonstrate the TCA cycle enzymatic steps, by using simple techniques to measure O(2) consumption and transmembrane potential (DeltaPsi). The several TCA cycle intermediates induce specific enzyme activities, which can be identified by respiratory parameters. Such a strategy is also used to evidence properties of the TCA cycle enzymes: ADP stimulation of isocitrate dehydrogenase and alpha ketoglutarate dehydrogenase; activation by citrate of downstream oxidation steps, e.g. succinate dehydrogenase; and regulation of the activity of isocitrate dehydrogenase by citrate action on the citrate/isocitrate carrier. Furthermore, it has been demonstrated that, in the absence of exogenous Mg(2+) , isocitrate dependent respiration favors the alternative oxidase pathway, as judged by changes of the ADP/O elicited by the inhibitor n-propyl galate. These are some examples of assays related with TCA cycle intermediates we can use in laboratory courses. PMID- 21638560 TI - Commentary: Problem-based learning and grade inflation. PMID- 21638561 TI - Test: Construction of genomic libraries. AB - Terms to be familiar with before you start to solve the test: genomic library, gel filtration, restriction endonuclease, plasmid, sticky ends, blunt ends, ligation, recombinant DNA, bacterium transformation, denaturation and renaturation of DNA, satellite DNA, telomere, centromere, unique and repetitive sequences. PMID- 21638562 TI - Commentary: U.S. Biotechnology Companies, foreign nationals, and U.S. graduate schools: The changing dynamics of accessibility to H-1B visa holders and possible impacts on graduate schools in the molecular life sciences and cognate disciplines. PMID- 21638563 TI - Virtual universities are not about to replace face-to-face teaching. PMID- 21638564 TI - Websites of note. PMID- 21638566 TI - IUBMB 50th Anniversary and the history of BAMBED. PMID- 21638567 TI - Products of isoprene metabolism. PMID- 21638568 TI - From metabolic pathways charts to animaps in 50 years. PMID- 21638569 TI - The early history of (32) P as a radioactive tracer in biochemical research: A personal memoir. AB - The concept of using radioactive isotopes as "tracers" of chemical conversions was conceived and developed by inorganic chemist Georg de Hevesy (Nobel Laureate in Chemistry 1943). In 1935, he began to apply the technique to various biological processes using (32) P, and his experiments revealed the dynamic character of physiology and metabolism. Following de Hevesy's lead, Samuel Ruben (University of California, Berkeley) exploited (32) P in 1937-38 for investigation of phospholipid metabolism. Between 1937 and 1940, Ruben and colleague Martin Kamen spearheaded tracer studies in various biological systems using (32) P, short-lived (11) C, and other radioactive isotopes. During this period, Kamen was responsible for cyclotron-produced radioactive tracers and was able to sustain de Hevesy's research by supplying him with (32) P. In 1940, Ruben and Kamen discovered long-lived (14) C, which later proved to be a very powerful tool for analysis of complex biochemical processes, such as the path of carbon in photosynthesis. Between 1946 and 1950, (32) P was used in studies of bacteriophage replication and photosynthetic metabolism. This memoir surveys the history of these early investigations. PMID- 21638570 TI - Facilitating understanding of the purine nucleotide cycle and the one-carbon pool: Part I: The purine nucleotide cycle. AB - Some metabolic processes are readily understood because they are circumscribed in metabolic pathways that have clearly identifiable beginning points, end products, and other features. Other metabolic pathways that do not appear to be straightforward pose difficulties for students. One such metabolic process, the purine nucleotide cycle, is discussed here. PMID- 21638571 TI - Allosteric modulation of monomeric proteins*. AB - Multimeric proteins (e.g. hemoglobin) are considered to be the prototypes of allosteric enzymes, whereas monomeric proteins (e.g. myoglobin) usually are assumed to be nonallosteric. However, the modulation of the functional properties of monomeric proteins by heterotropic allosteric effectors casts doubts on this assumption. Here, the allosteric properties of sperm whale myoglobin, human serum albumin, and human alpha-thrombin, generally considered as molecular models of monomeric proteins, are summarized. PMID- 21638572 TI - Evolution of innate immune systems*. AB - Innate immunity is the oldest form of defense and is found to some degree in all species. It predates the adaptive immune system, consisting of antibodies, B cells, T cells, and the major histocompatibility antigens. These are found only in higher vertebrates and have been the focus of the majority of immunological research, particularly in mice and humans, over the years. Knowledge of immunity in lower vertebrate and invertebrate species is now increasing rapidly, shedding light on the evolution of immunity and in many cases adding to our understanding of the mammalian system. Several recurring structural, genetic, and developmental mechanisms are common features in these processes in both the cellular and the molecular aspects of innate immunity. PMID- 21638573 TI - Vitamin E: Textbooks require updating. AB - Vitamin E was discovered in 1922, but in more than 60 years many new findings have added to the early one. However, its description in many textbooks has not been proportionally updated. The result is an inaccurate, incomplete, and often incorrect description of the function of this vitamin. In some other textbooks, vitamin E is absolutely neglected. In almost all books the existence of vitamin E deficiency diseases is ignored. Recent findings on vitamin E molecular functions and its related diseases are described in this article. PMID- 21638574 TI - Concepts of protein sorting or targeting signals and membrane topology in undergraduate teaching*. AB - The process of protein biogenesis culminates in its correct targeting to specific subcellular locations where it serves a function. Contemporary molecular and cell biology investigations often involve the exogenous expression of epitope- or fluorescent protein-tagged recombinant molecules as well as subsequent analysis of protein-protein interactions in vitro and in vivo. Fundamental knowledge of targeting signals that direct a polypeptide to various organelles or membrane domains is essential for the proper design of such recombinant molecules. A fundamental concept of membrane compartmentalization is also often useful for the interpretation of the preliminary results of interaction screens. Knowledge in targeting signals and post-translational dynamics of proteins should therefore be given sufficient emphasis in an undergraduate biochemistry or molecular biology curriculum. Such knowledge is essential, particularly for undergraduates or fresh graduates embarking on research projects in a cell and molecular biology laboratory. PMID- 21638575 TI - Molecular modeling of heme proteins using MOE: Bio-inorganic and structure function activity for undergraduates*S. AB - A biochemical molecular modeling project on heme proteins suitable for an introductory Biochemistry I class has been designed with a 2-fold objective: i) to reinforce the correlation between protein three-dimensional structure and function through a discovery oriented project, and ii) to introduce students to the fields of bioinorganic and coordination chemistry. Students are asked to identify several unknown heme proteins based on a careful analysis of covalent and noncovalent interactions at the active site of each protein, focusing on amino acid reactivity and H-bonding networks. Starting with the three-dimensional crystal structures of four unknown proteins, students isolate and examine the coordination environment of the iron center in order to predict the relative reactivity toward dioxygen (O(2) ) or hydrogen peroxide (H(2) O(2) ). The central question of the project is to determine how the same iron protoporphyrin IX cofactor can be used by four different proteins to carry out diverse reactions, from electron transfer, to reversible oxygen binding to hydrogen peroxide activation. Pedagogical reasons for implementation of this biomolecular discovery based activity and student evaluations are discussed. In addition to developing many of the three-dimensional visualization skills needed to successfully learn biochemistry, students also learn to use the versatile MOE molecular modeling program (Molecular Operating Environment), become familiar with metalloprotein reactivity, and are introduced to computational biochemistry research. PMID- 21638576 TI - Developing students' awareness of public understanding of science: A matter of survival? AB - Biochemists and life scientists in general are now encountering a paradox regarding the public image of their work; society demands and encourages pursuit of more knowledge but at the same time is suspicious and fears the novelties it brings. At this historical moment, we are experiencing a delicate balance between the birth of a new era and a downsizing the status of biochemistry. The future can go either way, and it is up to scientists to become deeply involved to make the first scenario overturn the second. This task demands awareness and engagement in scientific communication and the promotion of science. However, the work ahead demands continuous commitment for more than one generation. If we want to leave a legacy of a well regarded and prosperous science to our students, we must let them know why public understanding is crucial, indeed vital, and show them how to cope with and actually embrace it. This may actually be a survival skill for them! PMID- 21638577 TI - Replacing lectures in conventional university courses by text-based flexible learning can be a rewarding experience for the lecturer and students. AB - For nearly two decades, a proportion of lectures in certain modules in conventional undergraduate diploma and degree courses has been replaced by text based flexible learning. This flexible learning program has satisfied the various quality assurance procedures in place within the university throughout the period of implementation. Evaluation of students' performance and perceptions demonstrated that the text-based flexible learning program was an effective alternative to lectures. The advantages and disadvantages of this flexible learning program from the lecturer's perspective are discussed. PMID- 21638578 TI - Commentary: Access to the scientific and medical literature in the developing world. PMID- 21638579 TI - Hinari and Agora: Providing access for researchers in developing countries. AB - This article briefly describes two schemes, Health InterNetwork Access to Research Initiative (HINARI) and Access to Global Online Research in Agriculture (AGORA), which enable up to date scientific and clinical literature to be distributed via the World Wide Web to individuals in developing countries at zero or minimal cost as a result of agreements between publishers. PMID- 21638580 TI - Trends in funding science education, 1994-2004*. AB - This review looks at the changes over the last 10 years in priorities guiding funding decisions in science education. A general survey of trends in funding both graduate and undergraduate education are covered, followed by a description of specific National Science Foundation initiatives. Understanding these changes should help proposal writers put their current submissions into context and better reply to the call for proposals from various agencies. PMID- 21638581 TI - Poem: A genetic ode, or a melan coli tale. PMID- 21638582 TI - Poem: PC arrghh! PMID- 21638583 TI - Isolation of Caenorhabditis elegans genomic DNA and detection of deletions in the unc-93 gene using PCR. AB - PCR, genomic DNA isolation, and agarose gel electrophoresis are common molecular biology techniques with a wide range of applications. Therefore, we have developed a series of exercises employing these techniques for an intermediate level undergraduate molecular biology laboratory course. In these exercises, students isolate genomic DNA from the nematode Caenorhabditis elegans and use PCR to detect deletions in the C. elegans unc-93 gene. In advance of the exercises, wild-type and three different unc-93 deletion mutant strains are grown, harvested, and frozen by the instructor. In one approach, students isolate genomic DNA from each strain using a genomic DNA isolation kit and use agarose gel electrophoresis to analyze the DNA and to estimate its concentration. PCRs using primers directed to two different regions of the unc-93 gene are carried out on the genomic DNA from wild-type and mutant strains, and the PCR products are analyzed by agarose gel electrophoresis. Students analyze the gel to determine the approximate location and size of deletions in the three mutant strains. Alternatively, students may lyse single nematodes and carry out PCR in one laboratory session. These exercises should be easily adaptable to detection of well characterized deletions in any organism. PMID- 21638584 TI - Commentary: Changing minds with "trick" questions. PMID- 21638585 TI - Analysis of DNA replication in plant cells*. AB - Terms to be familiar with before you start to solve the test: cell division, phases of the cell cycle, phases of mitosis, chromosomes, chromatids, [(3) H]thymidine, pulse labeling, autoradiography. PMID- 21638586 TI - Commentary: The biotechnology landscape: I. The U. S. Biotechnology centers. PMID- 21638587 TI - Commentary: Curtis bonk and teaching and administering in stormy times. PMID- 21638588 TI - Websites of note. PMID- 21638591 TI - Exploring protein function and evolution using free online bioinformatics tools. PMID- 21638592 TI - The use of MERLOT in biochemistry and molecular biology education. AB - The referatory, Multimedia Educational Resources for Learning and Online Teaching (MERLOT), contains links to 1300 electronic teaching resources in biology and chemistry. Approximately 20% have been peer reviewed, and most have user comments or assignments attached. In addition to being a source of educational resources, the MERLOT project seeks to strengthen the educational community by providing peer review and recognition of quality electronic learning objects. PMID- 21638593 TI - Digital learning material for student-directed model building in molecular biologyS. AB - The building of models to explain data and make predictions constitutes an important goal in molecular biology research. To give students the opportunity to practice such model building, two digital cases had previously been developed in which students are guided to build a model step by step. In this article, the development and initial evaluation of a third digital case is described. It concerns the selection of bristles during Drosophila development. To mimic a real research situation in a more realistic way, students are given much more freedom while building their models and can thus follow their own model-building approach. At the same time, however, students are provided with a sufficient amount of support to ensure that they can build their models without the requirement of intensive supervision. PMID- 21638594 TI - Teaching intermediary metabolism linearly doesn't work. AB - Despite the fact that knowledge of the major biochemical metabolic pathways is essential to understanding the pathophysiology, clinical presentation, and management of many human diseases, there is disagreement among medical educators regarding the relevance of intermediary metabolism to the practicing physician and the expectations for medical students to master the various metabolic pathways. One basis for the widespread cynicism that exists regarding the need to teach medical students metabolic reactions and pathways derives from the fact that too often these aspects of biochemistry are taught as if they were isolated, unrelated, and independent bits of information. We use medium-chain acyl-CoA dehydrogenase deficiency as an example of how it is both possible and fruitful to teach two important but seemingly unrelated pathways in an integrated fashion and in the context of a specific clinical problem. This example illustrates how imbedding multiple metabolic pathways in a realistic clinical scenario and demonstrating the intimate interrelatedness of those pathways can provide the insight and understanding that are required for informed and rational diagnostic and therapeutic decision making. This article concludes with a series of practical and realistic recommendations for improving the teaching of intermediary metabolism. PMID- 21638595 TI - A docking interaction study of the effect of critical mutations in ribonuclease a on protein-ligand binding. AB - Enzymes with ribonucleolytic activity play a pivotal role in gene expression and cellular homeostasis by altering the levels of cellular RNA. Ribonuclease A has been the most well studied of such enzymes whose histidine residues (His(12) and His(119) ) play a crucial role in the catalytic mechanism of the protein. The ligands chosen for this study, 2'CMP and 3'CMP, act as competitive substrate analog inhibitors of this enzyme. Using molecular graphics software freely available for academic use, AutoDock and PyMol, we demonstrate that substitution of either histidine residue by alanine causes marked changes in the distances between these critical residues of the enzyme. The ligands in the docked conformation (particularly on mutation of His(119) to Ala) compensate for the altered free energy and hydrogen bonding abilities in these new protein-ligand complexes. PMID- 21638596 TI - Simplifying structure analysis projects with customizable chime-based templates*. AB - Structure/function relationships are fundamental to understanding the properties of biological molecules, and thus it is imperative that biochemistry students learn how to analyze such relationships. Here we describe Chime-based web page templates and tutorials designed to help students develop their own strategies for exploring macromolecular three-dimensional structures like those on our course website. The templates can easily be customized for any structure of interest, and some templates include a Command Entry Line and a Message Recall Box for more refined macromolecular exploration using RasMol/Chime image modification commands. The tutorials present students with an integrated overview of the image modification capabilities of the Chime plug-in and its underlying RasMol-based command structure as accessed through the Command Entry Line. The tutorial also illustrates how RasMol/Chime command syntax addresses specific formatted structural information in a standard Protein Data Bank file. Judging by the high quality of structure-based presentations given by students who have used these templates and tutorials, it appears that these resources can help students learn to analyze complex macromolecular structures while also providing them with convenient tools for creating scientifically meaningful and visually effective molecular images to share with others. (The templates, tutorials, and our course website can be viewed at the following URLs, respectively: tutor.lscf.ucsb.edu/instdev/sears/biochemistry/presentations/demos-downloads.htm, tutor.lscf.ucsb.edu/instdev/sears/biochemistry/tutorials/pdbtutorial/frontwindow. tml, and tutor.lscf.ucsb.edu/instdev/sears/biochemistry/.). PMID- 21638598 TI - Poem: Ethanol, my children! PMID- 21638597 TI - Modular laboratory courses: An alternative to a traditional laboratory program. AB - Our modular laboratory teaching program is characterized by two major features. First, each course is taught independently and not linked with a particular lecture course. Second, each course is designed to be completed within one-half semester or less. The modular organization has allowed incorporation of the latest technology, reduction of class sizes, and a greater number of choices to permit students to pursue their own interests. Full-time laboratory teaching personnel work together on the organization of the program as well as design and teach individual courses. The program has distinct and separate instructional goals, namely introduction, reinforcement, and advancement of fundamental scientific capacities. Such focus has facilitated efforts to coordinate laboratory courses within the biological sciences and among other departments in natural sciences and engineering, raising expectations as students progress through a program. PMID- 21638599 TI - Use of solid phase extraction in the biochemistry laboratory to separate different lipids. AB - Solid-phase extraction (SPE) was used to demonstrate how various lipids and lipid classes could be separated in a biochemistry laboratory setting. Three different SPE methods were chosen on their ability to separate a lipid mixture, consisting of a combination of a either a fatty acid, a triacylglycerol, a mono- or diacylglycerol, phospholipid, cholesterol, or cholesteryl ester into distinct lipid fractions. These mini-scale SPE methods used aminopropyl-bonded silica columns or silica Sep-Pak cartridges suitable for completion in a 2- to 3-h time block. Thin-layer chromatography of the isolated lipid fractions was used to show separation characteristics and to evaluate the utility and reproducibility of each SPE method. PMID- 21638600 TI - Commentary: Generating discussion during examinations. PMID- 21638601 TI - Can problem-based learning address content and process?*. AB - Problem-based learning was used to address the learning process and traditional content in a one semester nonmajors biochemistry course. Three different pedagogical strategies were evaluated in three versions of the course. The first version exclusively utilized literature-based learning (analysis of primary literature articles in small groups) with almost no lectures from the instructor. Classroom assessment addressed the research process, not content. In the second version, literature-based learning was supplemented with a 75-min lecture by the instructor before each article was analyzed as well as additional lectures as deemed necessary. Classroom assessment addressed process (70%) over content (30%). The third version was lecture-based, with the emphasis on traditional content; lectures were supplemented with literature analysis. Classroom assessment emphasized content, but about 10% of each exam focused on the research process. Student evaluations of the three versions of the course showed no significant difference in opinions about the format of the course. Postcourse interviews with students 1) confirmed previous studies that showed that the literature-based format greatly increased the involvement of students in their own learning, 2) showed that literature-based learning is compatible with an equal focus on content and process, and 3) showed that supplementing lecture based teaching with literature-based learning does not empower students to manage their own learning, although student response to this instructional method was positive. PMID- 21638602 TI - Molecular analysis of the causes of down syndrome in a family. AB - Terms to be familiar with before you start to solve the test: Down Syndrome (trisomy 21), karyotype, chromosome translocation, mosaicism, minisatellites, Southern blotting, DNA fingerprinting, nondisjunction, first and second meiotic division, homologous chromosomes, sister chromatids. PMID- 21638603 TI - Commentary: Wiki: The fast way to collaborative authoring. PMID- 21638604 TI - Achieving due recognition for web scholarship: Experience at a structural biology education website. AB - Peer-evaluation committees are now faced with assessing the scholarly value of World Wide Web resources such as tutorials, databases, dictionaries, and computation services. How should these activities be weighted in comparison with publications in research or education journals or presentations at meetings? Clearly, nontraditional scholarship requires nontraditional documentation. In this article, I tell how I have documented my structural biology web resources for peer evaluation and how my colleagues have responded. PMID- 21638605 TI - Websites of note. PMID- 21638607 TI - Comparison of student performance in cooperative learning and traditional lecture based biochemistry classes. AB - Student performance in two different introductory biochemistry curricula are compared based on standardized testing of student content knowledge, problem solving skills, and student opinions about the courses. One curriculum was used in four traditional, lecture-based classes (n = 381 students), whereas the second curriculum was used in two cooperative learning classes (n = 39 students). Students in the cooperative learning classes not only performed at a level above their peers in standardized testing of content knowledge and in critical thinking and problem-solving tasks (p < 0.05), but they also were more positive about their learning experience. The testing data are in contrast to much of the medical school literature on the performance of students in problem-based learning (PBL) curricula, which shows little effect of the curricular format on student exam scores. The reason for the improvement is undoubtedly multifactorial. We argue that the enhancement of student performance in this study is related to: 1) the use of peer educational assistants, 2) an authentic PBL format, and 3) the application of a multicontextual learning environment in the curricular design. Though educationally successful, the cooperative learning classes as described in this study were too resource intensive to continue; however, we are exploring incorporation of some of the "high context" aspects of the small-group interactions into our current lecture-based course with the addition of on-line PBL cases. PMID- 21638608 TI - Teachers as learners in a cooperative learning biochemistry class. AB - Upper level college students majoring in biochemistry at the University of New Mexico have the opportunity to participate in an advanced biochemistry course entitled "Biochemistry Education." This course introduces theories of teaching and learning, provides opportunities for participation in course organization, design, and assessment strategies, and requires practice in lecturing, exam writing, and grading. One component of this course required the biochemistry majors to act as educational assistants, leading problem-based learning sessions in a cooperative learning introductory survey biochemistry course for nonmajors. Problem-based learning scenarios used in this course were based on real-life biochemistry problems. As a result of their participation, the educational assistants increased their understanding of the biochemistry principles, gained an appreciation for the difficulty of the job of a "good teacher," developed new approaches to their own learning, and became more confident speakers. The participating biochemistry faculty were also positively affected by the collaborative approach they were attempting to model for the two sets of students and realized the benefits of truly cooperative team teaching. PMID- 21638609 TI - SigrafW: An easy-to-use program for fitting enzyme kinetic data. AB - SigrafW is Windows-compatible software developed using the Microsoft(r) Visual Basic Studio program that uses the simplified Hill equation for fitting kinetic data from allosteric and Michaelian enzymes. SigrafW uses a modified Fibonacci search to calculate maximal velocity (V), the Hill coefficient (n), and the enzyme-substrate apparent dissociation constant (K). The estimation of V, K, and the sum of the squares of residuals is performed using a Wilkinson nonlinear regression at any Hill coefficient (n). In contrast to many currently available kinetic analysis programs, SigrafW shows several advantages for the determination of kinetic parameters of both hyperbolic and nonhyperbolic saturation curves. No initial estimates of the kinetic parameters are required, a measure of the goodness-of-the-fit for each calculation performed is provided, the nonlinear regression used for calculations eliminates the statistical bias inherent in linear transformations, and the software can be used for enzyme kinetic simulations either for educational or research purposes. Persons interested in receiving a free copy of the software should contact Dr. F. A. Leone. PMID- 21638610 TI - Teaching microbial physiology using glucose repression phenomenon in baker's yeast as an example. AB - The yeast Saccharomyces cerevisiae has been used by human beings since ancient times for its ability to convert sugar to alcohol. Continual exposure to glucose in the natural environment for innumerable generations has probably enabled S. cerevisiae to grow in fermentative mode on sugars by switching off the genes responsible for respiration even under aerobic conditions. This phenomenon is referred to as the Crabtree effect. The present review focuses on glucose repression in S. cerevisiae from a physiological perspective. Physiological studies presented involve batch and chemostat experiments of the wild type and a mutant that lacks a trait partially responsible for the fermentative behavior. Various undergraduate student exercises have been (and can be) formulated to illustrate the concept of glucose repression. PMID- 21638611 TI - DNA replication and transcription: An innovative teaching strategy. AB - First-year students in genetics at the University of KwaZulu-Natal, South Africa, attend two general biology modules, one in each semester. Teaching involves four formal lectures per week of 45 min each, one 3-h practical, and one lecture period tutorial. These students, graduating from secondary education, are well schooled in rote leaning but are limited in critical thinking and find assessment questions belonging to the higher levels of Bloom's taxonomy difficult. All students attend the formal lectures together, up to 300 students, whereas for the tutorials they are grouped into small groups, no more than 40 students in a tutorial class, allowing for innovative teaching strategies. Students find the processes of DNA replication and transcription difficult because of the sequential steps involved in the processes together with limitations imposed by the enzymes involved. Furthermore, they find the significance and relationships between the different components of the processes very difficult. A tutorial was developed in which students are requested to demonstrate replication with line drawings, which are then used in various iterations of transcription. The tutorial is administered in the presence of a tutor that guides the step by step execution of the tutorial while stimulating active participation. In the past 2 years, the presentation of this and other similar tutorials in genetics has improved overall class performance on average by 15%. Furthermore, students seem to display a greater retention from the first year to the second, which was previously rather limited. A survey among first-year students revealed that the implementation of this tutorial facilitated studying and recall by helping students to organize thoughts, picture the sequence of events, understand fundamental concepts, and create a feeling of confidence. PMID- 21638612 TI - The mitochondrial phosphate-to-oxygen ratio is not an integer. AB - For many years, mitochondrial phosphate-to-oxygen ratios were believed to be integral values: 3 for ATP/NADH and 2 for ATP/FADH(2) . The chemiosmotic theory has demonstrated that this need not be the case because coupling sites do not directly synthesize ATP. Coupling between coupling site redox reactions and ATP synthesis occurs via proton pumping and proton flow, respectively. Most recent measurements support phosphate-to-oxygen ratios of ~2.5 ATP/NADH and 1.5 ATP/FADH(2) ; these values match those expected from chemiosmotic calculations. Faculty members who teach metabolism are encouraged to reflect these developments in their lecture notes. PMID- 21638614 TI - Poem: Organic matchmaker. PMID- 21638613 TI - Poem: A is for alanine. AB - The poem below presents a brief review of amino acid abbreviations and structures. Among the featured facts are the following: histidine has a pK(a) close to the normal pH of the cytosol and therefore is an effective intracellular buffer, the R group of isoleucine is hydrophobic, and many protein kinases add a phosphate at the -OH site of tyrosine. This work is just one of many science songs and poems that may be used for educational purposes. A comprehensive database of over 2,000 songs is available online at www.science groove.org/MASSIVE/. PMID- 21638615 TI - A series of enzymology-based experiments designed to mimic an applied research project. AB - Four mini-practicals are described in which the effects of temperature and pH on phytase activity are assessed, as well as the enzyme's thermostability and the effect upon stability of simulated digestive tract conditions. Phytase is routinely incorporated into monogastric animal feed to ameliorate the negative nutritional and environmental consequences of its substrate, dietary phytic acid. In addition to illustrating selected basic concepts in enzymology, the combined experiments allow the students to determine the suitability of the test phytase for inclusion in animal feed. As such the practical mimics an applied research project and is particularly suited to biotechnology students undertaking courses in basic biochemistry. Students may be segregated into groups of 4, with each team member charged with undertaking one of the mini-experiments. In this way students are given individual responsibility and learn to work as part of an integrated research grouping. PMID- 21638616 TI - In Vitro synthesis and activity of reporter proteins in an Escherichia coli S30 extract system: An undergraduate experiment*. AB - This undergraduate laboratory experiment integrates multiple techniques (in vitro synthesis, enzyme assays, Western blotting) to determine the production and detection sensitivity of two common reporter proteins (beta-galactosidase and luciferase) within an Escherichia coli S30 transcription/translation extract. Comparison of the data suggests that luciferase is the more suitable reporter for this specific in vitro extract system. Simple modifications in the experimental design allow for flexibility in the use of materials and the time required to perform the study. Furthermore, extension into additional experiments and alternative techniques are also discussed. PMID- 21638617 TI - Commentary: Problems without answers. PMID- 21638618 TI - The use of immunoprecipitation and affinity chromatography for the purification of a specific protein*. AB - Terms to be familiar with before you start to solve the test: immunoprecipitation, affinity chromatography, Western blotting, cyclin-dependent kinases, immunoglobulins, Coomassie Blue staining. PMID- 21638619 TI - Commentary: There are many (biochemical) paths to the truth. PMID- 21638620 TI - Metabolic interrelationships software application: Interactive learning tool for intermediary metabolism*. AB - We developed and implemented the software application titled Metabolic Interrelationships as a self-learning and -teaching tool for intermediary metabolism. It is used by undergraduate medical students in an integrated organ systems-based and disease-oriented core curriculum, which started in our medical faculty in 2001. The computer program provides an interactive environment in which students learn to integrate the major metabolic pathways as well as their hormonal control mechanisms as far as they depend on nutritional status. Students can explore the time- and tissue-dependent changes in mammalian intermediary metabolism during a feeding-fasting cycle. Starting from a whole-body view of interorgan nutrient fluxes, the student can make excursions to individual organs and, from there, to increasing levels of molecular detail and to explanatory animations. The application is well received by students and staff. PMID- 21638621 TI - Websites of note. PMID- 21638623 TI - Computer programming and biomolecular structure studies: A step beyond internet bioinformatics. AB - This article describes the experience of teaching structural bioinformatics to third year undergraduate students in a subject titled Biomolecular Structure and Bioinformatics. Students were introduced to computer programming and used this knowledge in a practical application as an alternative to the well established Internet bioinformatics approach that relies on access to the Internet and biological databases. This was an ambitious approach considering that the students mostly had a biological background. There were also time constraints of eight lectures in total and two accompanying practical sessions. The main challenge was that students had to be introduced to computer programming from a beginner level and in a short time provided with enough knowledge to independently solve a simple bioinformatics problem. This was accomplished with a problem directly relevant to the rest of the subject, concerned with the structure-function relationships and experimental techniques for the determination of macromolecular structure. PMID- 21638624 TI - Lost in translation? Deconstructing science in the news through an inquiry-based course. AB - This report describes an experiment to introduce freshmen science students to inquiry-based learning. The overarching theme was the communication of scientific information to the public by the mass media. Students, working in groups, deconstructed news items (many dealing with basic biomedical issues) and assessed the veracity of statements with reference to published scientific literature. Working as individuals, they wrote news items about the work of researchers whom they had interviewed, who included among others biochemists, molecular biologists, physiologists, pharmacologists, neuroscientists, and nutritionists. This experience enabled freshmen to look critically at mass media, as well as the scientific literature. As this approach transcends disciplinary boundaries, it is relevant for educating future biochemists and molecular biologists. PMID- 21638625 TI - Screen capture on the fly: Combining molecular visualization and a tablet PC in the biochemistry lecture*. AB - The biochemistry lecture is often the place where students receive the greatest exposure to the structural nature of biomolecules. The use of molecular visualization software and web-based animated tutorials has enhanced the way instructors teach this important area of structural biology. Using the software and tutorials in class is an excellent way to teach the diverse structural motifs found in biomolecules; however, integrating the computer-based molecular renderings shown in class into lecture notes is challenging. This report describes how incorporating a tablet PC into the biochemistry lecture can be used in conjunction with molecular visualization software to create a rich set of lecture notes. The pedagogical tools associated with a tablet PC make it an attractive addition to the biochemistry lecture, which usually has significant audio and visual learning components. PMID- 21638626 TI - The postage stamps of analogia. PMID- 21638627 TI - Reconsideration of the significance of substrate-level phosphorylation in the citric acid cycle*. AB - For nearly 50 years, students of metabolism in animals have been taught that a substrate-level phosphorylation in the Krebs citric acid cycle produces GTP that subsequently undergoes a transphosphorylation with ADP catalyzed by nucleoside diphosphate kinase. Research in the past decade has revealed that animals also express an ADP-forming succinate-CoA ligase whose activity exceeds that of the GDP-forming enzyme in some tissues. Here I argue that the primary fate of GTP is unlikely to be transphosphorylation with ADP. Rather, two succinate-CoA ligases with different nucleotide specificities have evolved to better integrate and regulate the central metabolic pathways that involve the citric acid cycle. The products of substrate-level phosphorylation, ATP and/or GTP, may represent a pool of nucleotide that has a different phosphorylation potential than the ATP made by oxidative phosphorylation and may be channeled to meet specific needs within mitochondria and the cell. Further research is needed to determine the applicable mechanisms and how they vary in tissues. PMID- 21638628 TI - A web-based genetic polymorphism learning approach for high school students and science teachers*. AB - Variation and polymorphism are concepts that are central to genetics and genomics, primary biological disciplines in which high school students and undergraduates require a solid foundation. From 1998 through 2002, a web-based genetics education program was developed for high school teachers and students. The program included an exercise on using freely available bioinformatics tools on the Internet to detect single nucleotide polymorphisms in genomic DNA and gene based sequences to evaluate variation or polymorphism. Similar tools were also used to show the functional effect, if any, of the single nucleotide polymorphisms. A total of 25 science teachers and 60 students from high schools in Alabama and Virginia participated in the program that ranged from 2 to 4 weeks. Seventy percent of the teachers have now developed a web-based module to teach at least two lessons involving DNA variation and how it influences other disciplines, including evolution. Among former high school students, five are in Ph.D. programs in genetics or related subjects, and 80% are in medical school or in college in a biology or pre-med major. The exercise is simple to implement, and the cost is relatively low, requiring only a computer with an Internet connection. It also provides a foundation for introducing students to the theory of evolution, a concept that remains controversial in high school science curricula. Similar programs, if properly implemented, may result in fostering more interest in the biological sciences among prospective college students and ensure a good foundation in the pipeline for career biologists and scientists. PMID- 21638629 TI - Poem: Party of 5.55 * 10(1) mol/L. PMID- 21638630 TI - Song: En-er-gy (to the tune of "let it Be")*. PMID- 21638631 TI - Song: Glucagon is coming around (to the tune of "santa claus is coming to town")*. PMID- 21638632 TI - Faux mutagenesis: Teaching troubleshooting through controlled failure. AB - By shifting pedagogical goals from obtaining successful mutations to teaching students critical troubleshooting skills, it has been possible to introduce site directed mutagenesis into an undergraduate teaching laboratory. Described in this study is an inexpensive laboratory exercise in which students follow a slightly modified version of Stratagene's QuikChange site-directed mutagenesis kit to effect a single amino acid change in ribonuclease Sa. From the students' perspective, they are performing an authentic mutagenesis reaction. However, by judicious substitution of most of the reagents, the exercise has been made economically feasible for implementation in large classes while still providing students the opportunity to learn not only the underlying theory of site-directed mutagenesis but also a host of associated concepts such as transcription, translation, PCR, DNA methylation, restriction digests, transformations, and blue/white screening. Just as importantly, this exercise simulates a wide range of mutagenesis failures, allowing students the opportunity to troubleshoot an experiment by carefully analyzing the results of positive and negative controls, thus helping to develop analytical thinking skills in a way that simply would not happen if students' experiments "worked." PMID- 21638633 TI - A thin layer chromatography laboratory experiment of medical importance. AB - A thin layer chromatography experiment of medical importance is described. The experiment involves extraction of lipids from simulated amniotic fluid samples followed by separation, detection, and scanning of the lecithin and sphingomyelin bands on TLC plates. The lecithin-to-sphingomyelin ratio is calculated. The clinical significance of this number is discussed. Since this is a procedure that is often performed in medical laboratories, most of the supplies and materials for the experiment are commercially available in kit form. PMID- 21638634 TI - Commentary: Litmus test for biochemistry students. PMID- 21638635 TI - The effect of actinomycin D on RNA metabolism in human cells. AB - Terms to be familiar with before you start to solve the test: pre-mRNA (hnRNA); 45 S pre-rRNA; 28 S and 18 S rRNA; pulse-chase labeling; [(3) H]uridine; RNA isolation; separation methods; centrifugation methods; chromatography methods; tRNA; 5 S rRNA; RNA synthesis; RNA processing; RNA transport; polyadenylation. (1) The abbreviations used are: hnRNA, heterogeneous nuclear RNA; MCQ, multiple choice questions. PMID- 21638636 TI - Commentary: A case for expanding the scope of case studies in biotechnology education. PMID- 21638637 TI - Commentary: For and against learning management systems. PMID- 21638638 TI - Websites of note. PMID- 21638640 TI - The American Society for Biochemistry and molecular biology interest in education. PMID- 21638641 TI - Cytochrome P450 enzymes in drug metabolism and chemical toxicology: An introduction. AB - Cytochrome P450 (P450) enzymes include a family of related enzymes that are involved in metabolism of vitamins, steroids, fatty acids, and other chemicals. This review presents a brief historical overview of the discovery and characterization of P450 enzymes extending from intermediary metabolism to the fields of drug metabolism and toxicology. Introductions to P450 enzyme structure and function are also presented. The goals of this review are to 1) provide an introduction to a few of the many aspects of P450 research relating to humans, 2) introduce additional ways of thinking about metabolism, 3) provide some basic examples of P450 enzymology, and 4) provide applications to topics widely taught in undergraduate courses in biochemistry. PMID- 21638642 TI - One hundred years of American Women in biochemistry *. AB - The occasion of ASBMB's 100th anniversary provides an opportunity to consider the history and accomplishments of women biochemists over the past century. American women in biochemistry have ranged from the highly visible, such as Nobel Prize winners Gert Cori and Gertrude Elion, to those who contributed in relative obscurity to equally dramatic breakthroughs. Women with Ph.D.s in biochemistry established departments of public health, nutrition, and home economics when more prestigious academic departments were not open to them. As in all fields of science, women have made great progress in biochemistry in recent years, and ASBMB reflects that progress; the past, current, and incoming presidents of the Society are all women. The data and biographical information presented here should provide a basis for inclusion of more women in recounting the history of biochemistry. PMID- 21638643 TI - Thoughts on the teaching of metabolism. AB - Systems biology, metabolomics, metabolic engineering, and other recent developments in biochemistry suggest that future biochemists will require a detailed familiarity with the compounds and pathways of intermediary metabolism and their biochemical control. The challenge to the biochemistry instructor is the presentation of metabolic pathways in a manner that allows student creativity in learning the pathways and their components. One approach that does permit the use of problem solving for the study of metabolic pathways involves following the fate of (13) C, (14) C, or (15) N labels, presented originally in the structure of an important metabolic starting compound, through relevant metabolic pathways. This method allows the presentation and study of problems in which such an isotopic label can be traced through two or more metabolic pathways, thus illustrating how these pathways are interconnected. The understanding that all the pathways of intermediary metabolism are interconnected provides opportunities to discuss their metabolic control by such mechanisms as signaling, feedback inhibition, location in organelles, coenzyme levels, and coenzyme recycling rates. The method is illustrated by following the fate of (14) C labels through anaerobic glycolysis, gluconeogenesis, and fatty acid transport, beta-oxidation, and ketone body formation. Cholesterol biosynthesis and heme formation are used to show that presentations of long and complex pathways can demonstrate important biochemical concepts by following the fate of an isotopic label using only the most important intermediates. Problems based on tracing radioactive labels through one or more metabolic pathways allow the use of cooperative learning techniques. PMID- 21638644 TI - Middle/high school students in the research laboratory: A summer internship program emphasizing the interdisciplinary nature of biology. AB - We describe an eight-week summer Young Scientist in Training (YSIT) internship program involving middle and high school students. This program exposed students to current basic research in molecular genetics, while introducing or reinforcing principles of the scientific method and demonstrating the uses of mathematics and chemistry in biology. For the laboratory-based program, selected students from Baltimore City Schools working in groups of three were teamed with undergraduate research assistants at Morgan State University. Teams were assigned a project that was indirectly related to our laboratory research on the characterization of gene expression in Caenorhabditis elegans. At the end of the program, teams prepared posters detailing their accomplishments, and presented their findings to parents and faculty members during a mini-symposium. The posters were also submitted to the respective schools and the interns were offered a presentation of their research at local high school science fairs. PMID- 21638645 TI - The importance of visual literacy in the education of biochemists*. AB - Visualization is an essential skill for all students and biochemists studying and researching the molecular and cellular biosciences. In this study, we discuss the nature and importance of visualization in biochemistry education and argue that students should be explicitly taught visual literacy and the skills for using visualization tools as essential components of all biochemistry curricula. We suggest that, at present, very little pedagogical attention has been given to this vital component of biochemistry education, although a large diversity of static, dynamic, and multimedia visual displays continues to flood modern educational resources at an exponential rate. Based on selected research findings from other science education domains and our own research experience in biochemistry education, 10 fundamental guidelines are proposed for the promotion of visualization and visual literacy among students studying in the molecular and cellular biosciences. PMID- 21638646 TI - A mechanistic foundation for instructor-regulated collective learning. AB - The theory of memory and the ability to influence learning with conscious and unconscious factors, both internal and external, is reviewed. From a connectionist view of memory, the interactive compensatory model of learning is introduced, with an emphasis on the plastic components (motivation, prior knowledge, and metacognition) that can be regulated. Expert students, but not naive students, have a considerable ability to self-regulate their learning processes and improve performance in undergraduate courses. We propose that in a class situation, there are emergent opportunities for an instructor who understands the dynamics of the learning environment to provide regulation for collective learning of multiple students and improve group learning. There are three interfaces for instructor-regulated collective learning, and examples are provided for each to suggest how instructors may effectively provide regulation. PMID- 21638647 TI - Song: The histone song (to the tune of "flintstones")*. PMID- 21638648 TI - Song: The genome song (to the tune of "it's a small world"). PMID- 21638649 TI - Song: If you're molecular and know it, clap your hands (to the tune of "if you're happy and you know it, clap your hands")*. PMID- 21638650 TI - Song: The sound of glucose (to the tune of "a few of my favorite things")*. PMID- 21638651 TI - Song: B-DNA (to the tune of "YMCA")*. AB - Text in regular type to be sung (and ideally danced) by professor. Underlined text to be sung by everyone (loudly). PMID- 21638652 TI - An exercise to estimate differential gene expression in human cells. AB - The expression of genes in cells of various tissue types varies considerably and is correlated with the function of a particular organ. The pattern of gene expression changes in diseased tissues, in response to therapy or infection and exposure to environmental mutagens, chemicals, ultraviolet light, and ionizing radiation. To better understand the disease at the molecular level, the alterations in gene expression can provide clues to identify the cellular pathways involved. There are a wide variety of techniques that have been developed to study differential gene expression. Most of these methodologies are technically challenging. We developed an exercise to examine and quantify differential gene expression based on reverse transcription-PCR. Our goal was to have this technique successfully performed by college level students. This technique is easy to perform and does not require the use of radioactive substances. We examined the ionizing radiation-induced changes in gene expression in human cells. After exposure to ionizing radiation, total RNA was isolated at 4, 8, 12, and 24 h. The RNA was converted to cDNA and subjected to PCR amplification using gene-specific primers and commercially available beta-actin internal standards in a multiplex format. The differences in gene expression were quantified with freely available imaging software. This technique should have a wide application to investigate differential gene expression in a variety of organisms and under various experimental and treatment conditions. PMID- 21638653 TI - Identification of highly prized commercial fish using a PCR-based methodology. AB - We report a practical class designed for undergraduate students of Marine Sciences as a part of the Genetics course. The class can also be included in undergraduate studies of food technology. The exercise was designed to emphasize the application of molecular biology techniques to fish species authentication and traceability. After a simple and rapid protocol for DNA extraction, PCR was used to analyze variation in size of individual 5S ribosomal gene spacer sequences in eight commercially important fishes. The resulting fragments containing these sequences were separated on agarose gels and visualized under a UV lamp. Differences in fragment length can be used to differentiate between species. This method is particularly useful in the identification of larvae, eggs, and processed food including frozen and canned products. PMID- 21638654 TI - Energetic metabolism and biochemical adaptation: A bird flight muscle model. AB - The main objective of this class experiment is to measure the activity of two metabolic enzymes in crude extract from bird pectoral muscle and to relate the differences to their mode of locomotion and ecology. The laboratory is adapted to stimulate the interest of wildlife management students to biochemistry. The enzymatic activities of cytochrome c oxidase and lactate dehydrogenase are measured in pectoral muscle of black duck and ring-necked pheasant. The black ducks have a high cytochrome c oxidase/lactate dehydrogenase (LDH) ratio, which reflects high aerobic capacity required for sustained and long distance flight. The low cytochrome c oxidase/LDH ratio in ring-necked pheasants and high level of LDH activity suggest that this bird can only support short bursts of flight, which may be related to his strategy of predator avoidance. PMID- 21638655 TI - Commentary: Do we need to understand how the brain works to teach well? PMID- 21638656 TI - Test: Targeted manipulation of mitochondrial DNA in mammalian cells*. PMID- 21638657 TI - Using advanced problems in introductory courses: Some sample problems and why they work. AB - This article presents several typical problems used in an introductory course in molecular biology and discusses why the problems are effective at increasing learning. PMID- 21638658 TI - Commentary: Is there a science lesson in the headlines? Scientific fraud and fraudulent science. PMID- 21638659 TI - The development of biotechnology education in China*. AB - From the middle of the 20th century, Chinese scientists have been actively involved in biotechnology. However, biotechnology education in China is a relatively recent phenomenon. This subject has not been addressed at the undergraduate level in a serious way until recently. In the last decade, biotechnology education developed rapidly and reached a new level in Chinese universities. The Chinese scientific establishment is very much aware of the importance of biotechnology and has identified this subject as one of the priority areas. Some universities are taking positive steps toward enhancing biotechnology education. This article focuses on the emergence, as well as the problems and prospects, of biotechnology education in China. PMID- 21638660 TI - HPV vaccine development: A case study of prevention and politics. AB - An analysis of the development of human papillomavirus (HPV) vaccines requires a fundamental comprehension of the underlying science of the technology, as well as an appreciation for the business issues essential to successful commercialization. Students analyzing this topic must consider scientific challenges related to the development of these vaccines, as well as the implications for their commercialization. This case study describes the evolution of our understanding of HPV infection and its causative link to cervical cancer, the establishment of viable HPV vaccine candidates, and various issues related to HPV vaccine implementation. Study questions for use in generating class discussion are provided. PMID- 21638661 TI - Perspective: Lessons from the elephant that is biotechnology. PMID- 21638662 TI - Commentary: Effective practice with e-learning. PMID- 21638665 TI - Code of ethics of the International Union of Biochemistry and molecular biology*. PMID- 21638663 TI - Websites of note. PMID- 21638666 TI - The determination of the redox states and phosphorylation potential in living tissues and their relationship to metabolic control of disease phenotypes. AB - This paper reviews the development in the 1950s of methods to determine the redox states of the free [NAD(+) ]/[NADH] in cytoplasm of yeast by Helmut Holzer and Feodore Lynen and in rat liver by Theodore Bucher and Martin Klingenberg. This work was extended in the 1960s in the laboratory of Hans Krebs, where the use of basic thermodynamic and kinetic principles allowed the extension of this approach to the determination of the free mitochondrial [NAD(+) ]/NADH] in mitochondria and the redox state of the free NADP system in cytoplasm and mitochondria. This work also outlined the linkage between the redox states in the various couples to the phosphorylation state or the free [ATP]/[ADP][P(i) ] ratio, the central energy parameter of living cells. This work has since been extended to include other energy-linked systems including the gradients of inorganic ions between extra and intracellular phases of the cell and the redox state of the co-enzyme Q couple of mitochondria. This system of linked near-equilibrium redox and phosphorylation potentials constitutes a framework of primitive metabolic control that is altered in a number of disease phenotypes. The alteration of such disease phenotypes by substrate availability is discussed, as well as the importance of a thorough grounding in basic kinetics and thermodynamics in designing new therapies to normalize the metabolic abnormalities that are the proximate cause of many common and some rare diseases states. PMID- 21638667 TI - Bioinformatics for undergraduates: Steps toward a quantitative bioscience curriculum*. AB - We describe an innovative bioinformatics course developed under grants from the National Science Foundation and the California State University Program in Research and Education in Biotechnology for undergraduate biology students. The project has been part of a continuing effort to offer students classroom experiences focused on principles and reasoning, using a computer-assisted, problem-based learning model. Here we report on the course design, methods of assessment for the course and materials, and analysis of results obtained in initial offerings of the course. PMID- 21638668 TI - A new three-dimensional educational model kit for building DNA and RNA molecules: Development and evaluation*. AB - International specialized literature focused on research in biology education is sadly scarce, especially regarding biochemical and molecular aspects. In this light, researchers from this Centre for Structural Molecular Biotechnology developed and evaluated a three-dimensional educational model named "Building Life Molecules DNA and RNA." The development of the model and its evaluation as a potential tool in the teaching-learning process were based on a pilot study involving 226 learners and teachers. Questionnaires were elaborated, containing simple and objective questions, similar to those used in research on science teaching, to orient the evaluation process. Our results show that the model has high educational potential, aiding participants in their conceptual understanding of these molecular structures and their functions, DNA semiconservative replication, and RNA transcription. In addition, it was observed that this model leads students to critical associations of these concepts with actual scientific themes of molecular biology and biotechnology, such as cloning, transgenic organisms, and the genome. PMID- 21638669 TI - Teaching technical and professional skills using a laboratory exercise: A comparison of two methods of plasmid preparation. AB - This laboratory exercise encourages upper level biochemistry students to build and expand upon previously developed laboratory skills and knowledge as they conduct a comparison of two methods of plasmid preparation based upon cost, quality of product, production time, and environmental impact. Besides creating an environment that mimics a more realistic practice of science, there are several key learning objectives. First, students will learn to effectively plan and manage time so that they can meet a scientific goal. A second objective is for them to learn to use theory as a basis for understanding new and different technology since classroom exposure is necessarily limited. Third, they will learn to think critically and logically to solve problems, and they will communicate this in written format. All of these skills are particularly important in preparation for a scientist's professional life. PMID- 21638670 TI - Engagement, exploration, explanation, extension, and evaluation (5E) learning cycle and conceptual change text as learning tools. AB - The purpose of this study is to investigate the effects of the Engagement, Exploration, Explanation, Extension, and Evaluation (5E) learning cycle, conceptual change texts, and traditional instructions on 8th grade students' understanding of photosynthesis and respiration in plants. Students' understanding of photosynthesis and respiration in plants was measured using the test developed by Haslam and Treagust (F. Haslam, D. F. Treagust (1987) Diagnosing secondary students' misconceptions of photosynthesis and respiration in plants using a two-tier multiple choice instrument, J. Biol. Educ. 21, 203 211). The test was administered as pre-test and post-test to a total of 101 8th grade students in three intact classes of the same school located in an urban area. The classes were randomly assigned as control and experimental groups. Students in the first experimental group (n = 33) received 5E learning cycle instruction, students in the second experimental group (n = 34) received conceptual change text instruction, and students in the control group (n = 34) received traditional instruction. Attitudes toward science and pre-test scores were used as covariates. A quasi-experimental design utilizing the analysis of covariance showed a statistically significant difference between the experimental and control groups in the favor of experimental groups after treatment. However, no statistically significant difference between two experimental groups (5E versus conceptual change text instruction) was found. PMID- 21638671 TI - Song: The plasmid song (to the tune of "will the circle be unbroken"). PMID- 21638672 TI - Song: The mRNA song (to the tune of "YMCA"). PMID- 21638673 TI - Song: The battle hymn of biochemistry (to the tune of "the battle hymn of the republic")*. PMID- 21638674 TI - Song: We all need just a little ATP (to the tune of "yellow submarine")*. PMID- 21638675 TI - Song: N-A-D (to the tune of "penny lane")*. PMID- 21638676 TI - Using yeast to easily determine mitochondrial functionality with 1-(4,5 dimethylthiazol-2-yl)-3,5-diphenyltetrazolium bromide (MTT) assay. AB - Mitochondrial respiration is an important topic in biochemical courses, but at the same time it is a very arduous and challenging subject for the students to comprehend. In teaching laboratories, experiments are also difficult to perform because mitochondrial isolation as well as oxygen consumption determinations require expensive equipment, rarely present in Mexican teaching laboratories. For this purpose, we adapted the 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenyltetrazolium bromide (MTT) assay, usually done with cell lines, to corroborate mitochondrial respiration in yeast. We used the reduction of the MTT to learn and distinguish the respiratory yield between different substrates as well as the effects of respiratory inhibitors. This is an economical, easily done laboratory experiment that quantifies mitochondrial functionality and provides a good idea of the respiratory process. PMID- 21638677 TI - Molecular cloning and analysis of a DNA repetitive element from the mouse genome. AB - We report the development of a 3-week laboratory activity for an undergraduate molecular biology course. This activity introduces students to the practice of basic molecular techniques such as restriction enzyme digestion, agarose gel electrophoresis, cloning, plasmid DNA purification, Southern blotting, and sequencing. Students learn how to carry out a GenBankTM search as they are encouraged to compile most available information about their cloned sequence. The natural integration of the bench work with the use of the bioinformatics tools is considered a major advantage of this laboratory course. PMID- 21638678 TI - "Mini-array" transcriptional analysis of the Listeria monocytogenes lecithinase operon as a class project: A student investigative molecular biology laboratory experience*. AB - This report describes a molecular biotechnology-based laboratory curriculum developed to accompany an undergraduate genetics course. During the course of a semester, students researched the pathogen, developed a research question, designed experiments, and performed transcriptional analysis of a set of genes that confer virulence to the food-borne pathogen, Listeria monocytogenes. Gene fragments were amplified via PCR and utilized in "mini-arrays," a dot-blot-based format suitable for the simultaneous transcriptional analysis of multiple genes. The project provides exposure to a wide range of molecular techniques and can be easily modified for variations in class size. Data are generated at various steps of the process, allowing for student interpretation, troubleshooting, and assessment opportunities. PMID- 21638679 TI - Commentary: Questioning for deeper understanding in problem-based learning*. PMID- 21638680 TI - Two restriction endonucleases. AB - Terms to be familiar with before you start to solve the test: restriction endonucleases; palindromic sequences; blunt ends; cohesive ("sticky") ends; in vitro DNA synthesis; radioactive nucleotide precursors; template-primer complexes; DNA ligation. PMID- 21638681 TI - Commentary: Have we received the wakeup call? PMID- 21638682 TI - Commentary: Space, the final frontier. PMID- 21638685 TI - The wandering pathway to determining N to C synthesis of proteins: Some recollections concerning protein structure and biosynthesis. PMID- 21638683 TI - Websites of note. PMID- 21638686 TI - Tactile teaching: Exploring protein structure/function using physical models*. AB - The technology now exists to construct physical models of proteins based on atomic coordinates of solved structures. We review here our recent experiences in using physical models to teach concepts of protein structure and function at both the high school and the undergraduate levels. At the high school level, physical models are used in a professional development program targeted to biology and chemistry teachers. This program has recently been expanded to include two student enrichment programs in which high school students participate in physical protein modeling activities. At the undergraduate level, we are currently exploring the usefulness of physical models in communicating concepts of protein structure and function that have been traditionally difficult to teach. We discuss our recent experience with two such examples: the close-packed nature of an enzyme active site and the pH-induced conformational change of the influenza hemagglutinin protein during virus infection. PMID- 21638687 TI - Biomolecules in the computer: Jmol to the rescue. AB - Jmol is free, open source software for interactive molecular visualization. Since it is written in the JavaTM programming language, it is compatible with all major operating systems and, in the applet form, with most modern web browsers. This article summarizes Jmol development and features that make it a valid and promising replacement for Rasmol and Chime in the development of educational materials, as well as in basic investigation of biomolecular structure. The description is set up by comparison with the well known abilities of Rasmol and Chime. Jmol is suitable for molecular model display and analysis in biochemistry, molecular biology, organic and inorganic chemistry, crystallography, and materials science. PMID- 21638688 TI - Evolution of protein lipograms: A bioinformatics problem. AB - A protein lacking one of the 20 common amino acids is a protein lipogram. This open-ended problem-based learning assignment deals with the evolution of proteins with biased amino acid composition. It has students query protein and metabolic databases to test the hypothesis that natural selection has reduced the frequency of each amino acid specifically in the enzymes required for its biosynthesis. Student groups work in parallel on different amino acids and share strategies. Aside from content objectives that integrate knowledge of protein structure, function, synthesis, and evolution, this problem incorporates oral and written presentations, statistical analysis, and substantial decision making. The point of the problem described here is that a deficiency or absence of a particular amino acid in a protein may be more than a chance occurrence and may be driven by natural selection. The challenge is to demonstrate the difference. PMID- 21638689 TI - Cognitive development and the complexities of the undergraduate learner in the science classroom*. AB - Students' reactions to classroom learning and the mastery of science vary along a wide spectrum of attitudes and emotions. In particular, we argue here that how learners encounter and learn subject matter is a function of their level of cognitive development. We describe the stages of cognitive development based on the work of William Perry and demonstrate their relevance to the undergraduate science classroom. With examples drawn from biochemistry, we attempt to show that, depending on the student's developmental level, there will be different abilities to handle the range of assignments and activities s/he can expect to experience in the average classroom. The college science instructor can benefit from knowledge of these stages and can work through their implications to develop strategies and techniques to regulate collective student learning. PMID- 21638690 TI - A descriptive analysis of computer-assisted teaching and learning in molecular biological education. AB - The role and importance of computer-assisted teaching and learning in molecular biological-related education and research has been emphasized and pinpointed. In this study, some benefit viewpoints and discussion are provided for applying the computer-assisted teaching and learning more efficiently in the process of knowledge acquisition and information exchange. Meanwhile, several cultural and economic issues related to the role of computers in teaching and learning are addressed. PMID- 21638691 TI - Pentose phosphate and calvin cycles: Similarities and three-dimensional views*. AB - The main object of this work is to present simplified and three-dimensional views of the pentose phosphate and Calvin cycles, emphasizing their functional and chemical similarities. PMID- 21638692 TI - Student understanding of pH: "i don't know what the log actually is, i only know where the button is on my calculator". AB - In foundation biochemistry and biological chemistry courses, a major problem area that has been identified is students' lack of understanding of pH, acids, bases, and buffers and their inability to apply their knowledge in solving acid/base problems. The aim of this study was to explore students' conceptions of pH and their ability to solve problems associated with the behavior of biological acids to understand the source of student difficulties. The responses given by most students are characteristic of an atomistic approach in which they pay no attention to the structure of the problem and concentrate only on juggling the elements together until they get a solution. Many students reported difficulty in understanding what the question was asking and were unable to interpret a simple graph showing the pH activity profile of an enzyme. The most startling finding was the lack of basic understanding of logarithms and the inability of all except one student to perform a simple calculation on logs without a calculator. This deficiency in high school mathematical skills severely hampered their understanding of pH. This study has highlighted a widespread deficiency in basic mathematical skills among first year undergraduates and a fragmented understanding of acids and bases. Implications for the way in which the concepts of pH and buffers are taught are discussed. PMID- 21638693 TI - Song: Learn about the buffer (generic rap)*. PMID- 21638694 TI - Song: Enzymes speed reactions (to the tune of "we will rock you," by queen)*S. PMID- 21638695 TI - Song: Chymotrypsin (to the tune of "eight days a week," by the beatles)*. PMID- 21638697 TI - Song: The codon song (to the tune of "when I'm sixty-four")*. PMID- 21638696 TI - Song: The ribosome (to the tune of "America the Beautiful")*. PMID- 21638698 TI - A nitrogen balance experiment using simulated urine samples. AB - We describe an undergraduate laboratory experiment that combines the advantages of problem-based learning with the need for biochemistry students to become proficient in practical laboratory skills. It also avoids the need to obtain ethical approval for recruiting volunteers and eliminates any possible biosafety issues with the handling and disposal of large amounts of urine. Simulated human urine samples are prepared that contain urea, uric acid, and creatinine at concentrations that represent the levels expected in 2 liters of urine collected over 24 h from subjects on various protein diets or during different physiological states. The students measure the nitrogen-containing compounds in the "urine samples" using specific colorimetric assays and use the data they generate to derive knowledge about nitrogen balance and the excretory metabolism of amino acids. PMID- 21638699 TI - Human salivary alpha-amylase (EC.3.2.1.1) activity and periodic acid and schiff reactive (PAS) staining: A useful tool to study polysaccharides at an undergraduate level. AB - Health science education is presently in discussion throughout Europe due to the Bologna Declaration. Teaching basic sciences such as biochemistry in a health sciences context, namely in allied heath education, can be a challenging task since the students of preclinical health sciences are not often convinced that basic sciences are clinically valuable (J. R. Rudland, S. C. Rennie (2003) The determination of the relevance of basic sciences learning objectives to clinical practice using a questionnaire survey, Med. Educ. (Oxf.) 37, 962-965; E. C. Wragg (2003) How can we determine the relevance of basic sciences learning objectives to clinical practice?, Med. Educ. (Oxf.) 37, 948-949). Thus, nowadays teachers are compelled to use their imagination to be able to elaborate laboratory sessions aiming for the understanding of theoretical concepts that are also clinically related: in other words, basic concepts and skills that underlie the competencies demanded of the future health professional. In the present work, we describe a set of laboratory sessions implemented in the discipline of biochemistry, belonging to the first year of several courses of allied health professionals, which can also be implemented in other health sciences courses. These sessions focus on the characteristics and properties of carbohydrates. The exercises we propose include two different laboratory practical sessions based on a histopathological routine technique known as periodic acid and Schiff reactive that is currently used to detect sugar metabolic and tumor diseases (J. M. T. Rivera, C. T. Lopez, B. C. Segui (2001) Bioquimica Estructural: Conceptos y Tests, Tebar Flores, Madrid). The methodology described enables the demonstration of some biochemical properties of polysaccharides, namely animal and vegetable, and the catalytic activity of the human salivary alpha-amylase (EC.3.2.1.1) enzyme. A further comparison between alpha-amylase activity in vitro and in situ is also possible by the proposed methodology. Additionally, to this extent, a comparison between the results of the learning improvement that occurred after the implementation of this tool is presented. PMID- 21638700 TI - Phylogenetics exercise using inherited human traits. AB - A bioinformatics laboratory exercise based on inherited human morphological traits is presented. It teaches how morphological characters can be used to study the evolutionary history of humans using parsimony. The exercise can easily be used in a pen-and-paper laboratory, but if computers are available, a more versatile analysis can be carried out. The exercise introduces students to the basics of bioinformatics laboratory work, shows them possible problems associated in data acquisition, and most importantly, teaches them basic group working skills, such as the appropriate distribution of work. An open and student oriented approach, in which group work is designed to support the learning of individual students, was adapted. The exercise is devised in two sessions that cover collection of data, analysis using parsimony, and a combined matrix analysis in a parsimony framework. The exercise, or parts of it, has successfully been applied in bioinformatics courses intended for second to third year biology majors and in adult education. PMID- 21638701 TI - Commentary: Who was linus pauling? PMID- 21638702 TI - The yeast two-hybrid system. AB - Terms to be familiar with before you start to solve the test: transcription factors, enhancer, promoter, RNA polymerase, expression plasmid, terminator, histidine, transfection, cell culture, fusion proteins, RNA splicing. PMID- 21638703 TI - Commentary: Taking a stand for science. PMID- 21638704 TI - Commentary: Gender as a factor in multimedia usage: Some observations and a little advice to offer. PMID- 21638705 TI - Websites of note. PMID- 21638707 TI - Glyceroneogenesis, the pathway that almost wasn't. AB - "What seest thou else in the dark, backward abysm of time." Prospero in The Tempest As is true in all aspects of human endeavor, a scientific concept can appear before its time and remain unappreciated before events catch up with the concept. Such was the case of the discovery of glyceroneogenesis and the establishment of its biological importance; it took almost 40 years before the significance of this pathway became apparent and the concept of triglyceride recycling was understood by the scientific establishment. Even that may be stretching a point, because today glyceroneogenesis is hardly a household word. In this essay, we will tell the story of the discovery of glyceroneogenesis and the thought processes that led us to propose this pathway. We will also speculate on why the pathway was not more widely embraced by scientists working in lipid metabolism and why that may finally be changing. The reader is warned, however, that this story is a reconstruction of past events and, like all such attempts, suffers from the patina of nostalgia that inevitably covers all things resurrected from memory. Others may view things differently, but this is our story as we remember it. PMID- 21638708 TI - Commentary: Science education as mystagogy? PMID- 21638709 TI - A course designed for undergraduate biochemistry students to learn about cultural diversity issues. AB - Biology, biochemistry, and other science students are well trained in science and familiar with how to conduct and evaluate scientific experiments. They are less aware of cultural issues or how these will impact their careers in research, education, or as professional health care workers. A course was developed for advanced undergraduate science majors to learn about diversity issues in a context that would be relevant to them, entitled "Diversity Issues in Health Care: Treatment and Research." Learning objectives included: developing awareness of current topics concerning diversity issues in health care; learning how research is carried out in health care, including pharmaceutical research, clinical trials, and social research; and learning about health care practices. Lectures and projects included readings on laboratory and clinical research, as well as literature on legal, race, gender, language, age, and income issues in health care research and clinical practice. Exams, papers, and a service learning project were used to determine the final course grade. Assessment indicated student understanding of diversity issues was improved, and the material was relevant. PMID- 21638710 TI - Intrageneric primer design: Bringing bioinformatics tools to the class. AB - Bioinformatics is one of the fastest growing scientific areas over the last decade. It focuses on the use of informatics tools for the organization and analysis of biological data. An example of their importance is the availability nowadays of dozens of software programs for genomic and proteomic studies. Thus, there is a growing field (private and academic) with a need for bachelor of science students with bioinformatics skills. In consideration of this need, described here is a problem-based class in which students are asked to design a set of intrageneric primers for PCR. The exercise is divided into five classes of 1 h each, in which students use freeware bioinformatics tools and data bases available through the Internet. Besides designing the set of primers, the students will consequently learn the significance and use of the major bioinformatics procedures, such as searching a data base, conducting and analyzing sequence multialignment, comparing sequences with a data base, and selecting primers. PMID- 21638711 TI - Dishonesty in the biochemistry classroom laboratory: A synthesis of causes and prevention. AB - Although reports of academic cheating are abundant, there are relatively few papers in the literature that focus on cheating in the context of science courses and even fewer that address dishonest practices, such as "cooking" or fudging data, within the classroom laboratory. This paper briefly reviews the existing literature on academic dishonesty and explores two theories that can be used to explain why cheating occurs: (1) classroom goal structure and (2) attitudes of neutralization. We conclude with a discussion of the implications of these theories within the context of a biochemistry and molecular biology teaching laboratory. PMID- 21638712 TI - jAMVLE, A new integrated molecular visualization learning environment*. AB - A new computer-based molecular visualization tool has been developed for teaching, and learning, molecular structure. This java-based jmol Amalgamated Molecular Visualization Learning Environment (jAMVLE) is platform-independent, integrated, and interactive. It has an overall graphical user interface that is intuitive and easy to use. The application can be downloaded free from the internet at wabri.org.au/jamvle. A cohort of 28 third year undergraduate molecular biotechnology degree students evaluated the new application through an essay-style project. These were analyzed to identify themes expressed by students in the content of their evaluations. Most students were positive about the new jAMVLE learning environment, and five major benefits emerged from the analysis. In particular, the students perceived that jAMVLE has an appealing interface, is interactive, provides a useful integrated environment, is user friendly, and is an excellent learning tool. Overall, students found that the jAMVLE application stimulated their interest, was a more active learning environment, provided better guidance, and made learning fun. PMID- 21638713 TI - Song: I'm a little mitochondrion (to the tune of "I'm a Lumberjack and I'm ok")*. PMID- 21638714 TI - Song: The cell's lament (to the tune of "yesterday")*. PMID- 21638715 TI - Song: Fatty acids in our cells (to the tune of "halls of montezuma")*. PMID- 21638716 TI - Song: This biochemistry (to the tune of "my country 'tis of thee")*. PMID- 21638717 TI - Song: The Vegetarian's song (to the tune of "blowin' in the wind")*. PMID- 21638718 TI - Microarrays (DNA chips) for the classroom laboratory. AB - We have developed and optimized the necessary laboratory materials to make DNA microarray technology accessible to all high school students at a fraction of both cost and data size. The primary component is a DNA chip/array that students "print" by hand and then analyze using research tools that have been adapted for classroom use. The primary adaptation is the use of a simulated cDNA target. The low density DNA array we discuss here was used to demonstrate differential expression of several Arabidopsis thaliana genes related to photosynthesis and photomorphogenesis. The methods we present here can be used with any biological organism whose sequence is known. Furthermore, these methods can be adapted to exhibit a variety of differential gene expression patterns under different experimental conditions. The materials and tools we discuss have been applied in classrooms at West High School in Madison, WI. We have also shared these materials with high school teachers attending professional development courses at the University of Wisconsin-Madison. PMID- 21638719 TI - A simple assay to quantify metallothionein helps to learn about bioindicators and environmental health. AB - We present a laboratory practice aimed to clarify and discuss the concept of bioindicator molecules in environmental health. It has been implemented as a hands-on laboratory exercise in postgraduate courses in public health (in Brazil) and in a course entitled Biology Applied to Natural Resources Sustainability (in Spain) but can be useful for undergraduate studies. By simple, repeatable, and low cost spectrophotometry methodology, students detect minimal concentrations (nmol) of metallothionein in biological samples obtained from sites with various status of pollution. They can also expose experimental fish or mollusks to heavy metals in aquarium conditions and then detect metallothionein. This method is excellent for didactic purposes because it can be developed even in modest laboratories with basic equipment for molecular biology. PMID- 21638720 TI - Fluorescence analysis of sulfonamide binding to carbonic anhydrase. AB - A practical laboratory experiment is described that illustrates the application of fluorescence resonance energy transfer to the study of protein-ligand binding. The affinities of wild-type and mutant human carbonic anhydrase II for dansylamide were determined by monitoring the increase in ligand fluorescence that occurs due to energy transfer from tryptophan residues near the enzyme active site. In a subsequent experiment, the binding constant of azetazolamide, a weaker fluorophore but a stronger ligand, is measured by competition with dansylamide. This simple experiment introduces students to the widely used technique of fluorescence spectroscopy and to the determination of protein-ligand binding constants. PMID- 21638721 TI - Glycobiology, how to sugar-coat an undergraduate advanced biochemistry laboratory. AB - A second semester biochemistry laboratory has been implemented as an independent projects course at California State University, Sacramento since 1999. To incorporate aspects of carbohydrate biochemistry, or glycobiology, into our curriculum, projects in lectin isolation and purification were undertaken over the course of two semesters. Through this modification in course content, this class now offers a diverse, hands-on treatment of not only standard protein purification techniques but also carbohydrate techniques, specifically the study of carbohydrate-protein interactions through hemagglutination assays, a novel commercial assay known as the InstantTMChek assay, and the generation and use of appropriate affinity chromatography matrices. Throughout the semester, the students are in charge of all aspects of their projects, from planning to execution and completion. Specific examples of student projects are highlighted such that the breadth of protein-carbohydrate chemistry pursued in a 15-week semester can be appreciated. The feedback of the course was very favorable, indicating that the students came away with skills necessary for them to be successful in their future careers. Most importantly, however, aspects of glycobiology have now been incorporated effectively into a mainstream undergraduate biochemistry laboratory class. PMID- 21638722 TI - DNA fingerprint analysis of three short tandem repeat (STR) loci for biochemistry and forensic science laboratory courses. AB - We have devised and implemented a DNA fingerprinting module for an upper division undergraduate laboratory based on the amplification and analysis of three of the 13 short tandem repeat loci that are required by the Federal Bureau of Investigation Combined DNA Index System (FBI CODIS) data base. Students first collect human epithelial (cheek) cells using sterile buccal swabs and then utilize commercially available kits and materials to extract genomic DNA. This is followed by the PCR amplification of three specific short tandem repeat loci (i.e. CSF1PO, TPOX, THO1). Polyacrylamide gel electrophoresis is used to resolve the allelic bands associated with the three short tandem repeat loci, and the results are statistically analyzed in the context of human population genetics. In addition, DNA was collected from a family, and the children's allele sets were compared with those of the parents to help illustrate paternal and maternal relatedness. This module enables students to use the materials and methods employed by actual law enforcement agencies and therefore can be used for laboratory exercises in traditional biochemistry curricula as well as for the growing field of forensic science and education. PMID- 21638723 TI - Commentary: Teaching without bloom or piaget. PMID- 21638724 TI - The effect of alpha-amanitin on RNA polymerase II ubiquitination*. AB - Terms to be familiar with before you start to solve the test: RNA polymerase II, transcription, alpha-amanitin, protein ubiquitination, synchronization of cell cultures, affinity adsorption, SDS-polyacrylamide gel electrophoresis, Western blotting, protein phosphorylation, protein kinases, in vitro transcription, plasmid, promoter, general and regulatory transcription factors, autoradiography. PMID- 21638725 TI - Commentary: Mobile phones, a technology looking for a pedagogy. PMID- 21638726 TI - Websites of note. PMID- 21638727 TI - Marks' basic medical biochemistry: A clinical approach (second edition). PMID- 21638728 TI - The Strongest boy in the world: How genetic information is reshaping our lives. PMID- 21638730 TI - BAMBED Changes Publisher: Farewell, ASBMB; welcome, John Wiley. PMID- 21638731 TI - EZ-Viz, a tool for simplifying molecular viewing in PyMOL. AB - PyMOL is a molecular modeling program that can be used in a wide range of studies within the scientific community. Its ability to produce informative, detailed, stereoscopic images makes it a very powerful tool both in the laboratory and in the classroom. However, the PyMOL user interface is difficult to use, and the PyMOL User's Manual readily admits that "development has been focused on capabilities, not on ease-of-use for new users." To resolve this, we set out to develop a new interface for PyMOL that helps eliminate the need for command line interactions, making it a much more user-friendly interface. We have created a tabbed interface called EZ-Viz, which contains a series of buttons and entry fields that allow users to select given attributes of a molecule and make changes to it by clicking a button as opposed to issuing highly syntactical commands. Other features of the interface include a Chime/PyMOL command converter, nine preset molecular views, four molecular movies, and molecular sequence and hetero atom information, all at the click of a button. The ongoing and continued development of EZ-Viz could help to further the use of PyMOL, which we believe to be one of the most powerful molecular viewing programs currently available. PMID- 21638732 TI - Databases of metabolic pathways. AB - Studies of metabolism usually evoke memories of densely written wall charts of metabolic pathways of enormous complexity, such as the IUBMB-Nicholson metabolic pathways map or the Roche metabolic pathways wall chart. Advances in bioinformatics in the past decade have brought about a revolution in the way the information in metabolic pathways is viewed and managed. Today, metabolic pathways are stored in knowledge bases that can be queried, modified, copied, and extended with ease, precision, and speed. Moreover, the flexibility of the digital medium for the presentation of metabolic pathways surpasses the traditional printed form used in classical pathway maps. In digital presentations, the separation of the content from the visual presentation is the norm, implying that metabolic pathway maps can be generated dynamically from the information stored in the knowledge base. The most advanced metabolic pathways databases are freely available over the Internet, and this has important implications for modern studies of metabolic pathways in biochemistry and molecular biology. This study reviews two such databases: the Kyoto Encyclopedia of Genes and Genomes (KEGG) and the BioCyc collection of pathway/genome databases. The background information and the structure of these databases are presented, with the emphasis on BioCyc, which represents a global multicollaborative effort to unify the representation of metabolic pathways. PMID- 21638733 TI - Time, the forgotten dimension of ligand binding teaching. AB - Ligand binding is generally explained in terms of the equilibrium constant K(d) for the protein-ligand complex dissociation. However, both theoretical considerations and experimental data point to the life span of the protein-ligand complex as an important, but generally overlooked, aspect of ligand binding by macromolecules. Short-lived protein-ligand complexes may be unable to trigger further biological processes as signal transduction or internalization if such processes are relatively slow with respect to dissociation of the complex that initiated them. Protein-ligand complex life span depends on the first-order rate constant for the dissociation of the complex, K(off) , but this constant and its implications are generally not treated in textbooks. This report presents a brief discussion and some examples useful for teaching the importance of time in ligand binding by macromolecules in the context of a general biochemistry course. PMID- 21638734 TI - Growth of microbial populations. Mathematical modeling, laboratory exercises, and model-based data analysis. AB - This paper has arisen as a result of teaching Models in Biology to undergraduates of Bioengineering at the Gediminas Technical University of Vilnius. The aim is to teach the students to use a fresh approach to the problems they are familiar with, to come up with an articulate verbal model after a mental effort, to express it in rigorous mathematical terms, to solve (with the aid of computers) corresponding equations, and finally, to analyze and interpret experimental data in terms of their (mathematical) models. Investigation of microbial growth provides excellent possibilities to combine laboratory exercises, mathematical modeling, and model-based data analysis. Application of mathematics in this field proved to be very fruitful in getting deeper insight into the processes of microbial growth. The step-by-step modeling resulted in an extended model of the growth covering conventional "lag," "exponential," and "stationary" phases. In contrast to known models (differential equations that can be solved only numerically), the present model is expressed symbolically as a finite combination of elementary functions. The approach can be applied in other areas of modern biology, such as dynamics of various cellular processes, enzyme and receptor kinetics, and others. PMID- 21638735 TI - Song: Hark the sucrose (to the tune of "hark the herald angels sing"). PMID- 21638736 TI - Song: O little protein molecule (to the tune of "O little town of Bethlehem"). PMID- 21638737 TI - Song: God rest ye merry lipoproteins (to the tune of "god rest ye merry gentlemen"). PMID- 21638738 TI - Song: the E. coli song (to the tune of "rudolph the red-nosed reindeer"). PMID- 21638739 TI - Song: Biochemistry biochemistry (to the tune of "O christmas tree"). PMID- 21638740 TI - Analyzing ligand depletion in a saturation equilibrium binding experiment. AB - I present a proposal for a laboratory practice to generate and analyze data from a saturation equilibrium binding experiment addressed to advanced undergraduate students. [(3) H]Quinuclidinyl benzilate is a nonselective muscarinic ligand with very high affinity and very low nonspecific binding to brain membranes, which contain a high density of muscarinic receptors. These features allow the instructor to devote especial emphasis to evaluate ligand depletion, and therefore, stress the subtle but fundamental difference between total (added) ligand and free ligand concentration at equilibrium. PMID- 21638741 TI - Microarrays for undergraduate classes. AB - A microarray experiment is presented that, in six laboratory sessions, takes undergraduate students from the tissue sample right through to data analysis. The model chosen, the murine erythroleukemia cell line, can be easily cultured in sufficient quantities for class use. Large changes in gene expression can be induced in these cells by erythropoietic agents such as DMSO over a 72-h time period. Students isolate total RNA from control (0 h) and 72-h DMSO-treated murine erythroleukemia cells. From this, they synthesize a cDNA copy incorporating amino-allyl dUTP, which is then coupled to either a Cy5 or a Cy3 dye. Equal amounts of the two labeled cDNA samples are then applied to a standard cDNA microarray, which is then hybridized, washed, and scanned. Up- and down regulated genes are selected using an "in-house" user-friendly data base program. Quality control checks are included at various stages throughout the procedure and, as the process of erythropoiesis is well characterized, a number of erythroid sequences serve as internal controls on the validity of the array data. Through this experiment, students gain experience in a wide range of molecular biology techniques, the use of controls to check a multistep process, validation of results, and strategies to manage the large amount of data generated. Most importantly, it provides undergraduate students with an opportunity to carry out experiments using cutting edge techniques normally found only in research laboratories. PMID- 21638742 TI - Oxidative hemolysis of erythrocytes. AB - This exercise for students will allow them to simultaneously observe lipid peroxidation and consequent hemolysis of rat erythrocytes and the effect of sodium azide, a catalase inhibitor, on these processes. It will also demonstrate a protective action of antioxidants, the therapeutically used N-acetylcysteine and albumins present in plasma. PMID- 21638744 TI - Commentary: Gender, teaching reform, promotion, and tenure. PMID- 21638743 TI - Exploring the ubiquitin-proteasome protein degradation pathway in yeast. AB - This article describes an undergraduate biochemistry laboratory investigating the ubiquitin-proteasome pathway in yeast. In this exercise, the enzyme beta galactosidase (beta-gal) is expressed in yeast under the control of a stress response promoter. Following exposure to heat stress to induce beta-gal expression, cycloheximide is added to halt translation, and beta-gal degradation is monitored by measuring enzyme activity as a function of time. Students observe that an N-Ile-beta-gal variant with an N-terminal isoleucine has a significantly lower stability than wild-type beta-gal, whose N-terminal residue is methionine. This strong dependence of protein stability on the N-terminal residue is known as the "N-end rule." To corroborate the enzyme activity assay results, students perform denaturing protein electrophoresis and immunoblotting of lysates, observing that the time-dependent loss of enzyme activity is coincident with the disappearance of the beta-gal protein. PMID- 21638745 TI - Vectorial Transport in intestinal epithelial cells. AB - Terms to be familiar with before you start to solve the test: polarized cells, vectorial transport, apical and basolateral membrane, tight junction, occludin, transmembrane proteins, passive and active transport, glucose transporter, Na(+) /glucose symporter, Na(+) /K(+) ATPase, uniporter, antiporter, facilitated diffusion, lateral diffusion. PMID- 21638746 TI - Commentary: The "business" of the biosciences can be integrated into a biochemistry curriculum. PMID- 21638747 TI - Commentary: Mystagogy, the third way. PMID- 21638748 TI - Commentary: Mystagogy, the tertiary way. PMID- 21638749 TI - Websites of note. PMID- 21638751 TI - High-speed multicolor microscopy of repeating dynamic processes. AB - Images of multiply labeled fluorescent samples provide unique insights into the localization of molecules, cells, and tissues. The ability to image multiple channels simultaneously at high speed without cross talk is limited to a few colors and requires dedicated multichannel or multispectral detection procedures. Simpler microscopes, in which each color is imaged sequentially, produce a much lower frame rate. Here, we describe a technique to image, at high frame rate, multiply labeled samples that have a repeating motion. We capture images in a single channel at a time over one full occurrence of the motion then repeat acquisition for other channels over subsequent occurrences. We finally build a high-speed multichannel image sequence by combining the images after applying a normalized mutual information-based time registration procedure. We show that this technique is amenable to image the beating heart of a double-labeled embryonic quail in three channels (brightfield, yellow, and mCherry fluorescent proteins) using a fluorescence wide-field microscope equipped with a single monochrome camera and without fast channel switching optics. We experimentally evaluate the accuracy of our method on image series from a two-channel confocal microscope. PMID- 21638752 TI - Co-variation in frequency and severity of cardiovascular and skeletal defects in Sprague-Dawley rats after maternal administration of dimethadione, the N demethylated metabolite of trimethadione. AB - BACKGROUND: The anticonvulsant trimethadione is a potent inducer of ventricular septation defects, both clinically and in rodents. Teratogenicity requires its N demethylation to dimethadione, the proximate teratogen. It was previously demonstrated trimethadione only induced membranous ventricular septation defects in rat (Fleeman et al., 2004), and our present goal is to determine whether direct administration of dimethadione increases the incidence and severity of septation defects. METHODS: Pregnant Sprague-Dawley rats were divided into five groups and administered either distilled water (control) or four different regimens of dimethadione. The core treatment was 300 mg/kg dimethadione b.i.d. on gestation day 9, 10 with additional groups given one additional dose of dimethadione 12 hr earlier, 12 hr later or two additional doses 12 hr earlier and later. Caesarian sections occurred on gestation day 21 and fetuses were examined for standard developmental toxicity endpoints. RESULTS: The broadest dosing regimen yielded the highest incidence and the most severe heart and axioskeletal findings with a decrease in mean fetal body weight. The overall incidence of ventricular septation defects was 74%, of which 68% were membranous and 9% muscular. Outflow tract anomalies (17%) were also observed, as were malformations of the axioskeleton (97%), but not of the long bones, and of particular interest was the high incidence of sternoschesis. CONCLUSIONS: Unlike trimethadione, dimethadione induces more serious muscular septation defects that are believed to be more clinically relevant. This, when taken together with the high incidence of total septation anomalies suggests dimethadione is useful for the study of chemically induced ventricular septation defects. PMID- 21638753 TI - A diet of somatic cell nuclear transfer cloned-cattle meat produced no toxic effects on behavioral or reproductive characteristics of F1 rats derived from dams fed on cloned-cattle meat. AB - BACKGROUND: The composition and nutritional value of meat and milk derived from cloned animals and their progeny has not been demonstrated to be different from normal animals, but possible food consumption risks that might arise from unidentified hazards remain. In this study, we investigated the effects of somatic cell nuclear transfer cloned-cattle meat diet on the behavioral and reproductive characteristics of F1 rats derived from dams that were also fed on cloned-cattle meat. METHODS AND RESULTS: F1 rats were divided into five diet groups with their dams: commercial pellets (control), pellets containing 5% (N-5) and 10% (N-10) of normal-cattle meat, and diets containing 5% (C-5) and 10% (C 10) of cloned-cattle meat. In most cases, the cloned-cattle meat diet did not affect body weight and food consumption in both male and female F1 rats during 11 weeks, except for significantly higher body weight in both N-5 and N-10 (3-5 weeks, p<0.05 or p<0.01) and significantly higher food consumption in the both normal- and cloned-cattle meat groups (7-9 weeks, p<0.05 or p<0.01), as compared with the controls, respectively. We detected no signs of test substance-related toxicities on organ weights and behavioral characteristics (sensory reflex, motor function, and spatial learning and memory tests). Reproductive functions did not significantly differ among all examined rats (mating, fertility, and implantation). CONCLUSIONS: These behavioral and reproductive toxicity results suggest that there are no obvious food safety concerns related to cloned-cattle meat in these parameters. PMID- 21638754 TI - Juvenile animal studies and pediatric drug development: a European regulatory perspective. AB - During the workshop organized by ILSI/HESI on May 5-6, 2010 on the value of juvenile animal toxicity studies, the implementation of the European Pediatric Regulation and in particular the review process of the nonclinical part of the Pediatric Investigation Plan (PIP) were described. A PIP is intended to outline the development of a medicinal product in the pediatric population (i.e. quality, safety, efficacy of the medicine and timing of studies); it is reviewed and agreed by the Pediatric Committee (PDCO) of the European Medicines Agency (EMA). The Nonclinical Working Group (NcWG) supports the PDCO in the review process of the nonclinical part of a PIP and is composed of members from the PDCO, the EMA Safety Working Party, additional experts from national competent authorities and the FDA. This article summarizes the NcWG review process and outcomes of 97 approved or ongoing PIPs, from the establishment of the NcWG in November 2008 to May 2010, as presented during the workshop. Juvenile animal studies were proposed by the applicant in 33% or required by the NcWG in 26% of the PIPs. The requirements were mainly motivated by concerns regarding potential developmental toxicities, in view of the young age of the pediatric population to be investigated, the lack of knowledge concerning the maturation of the pharmacological target, the lack of sufficient (non)clinical data, observed toxicities in the adult (non)clinical studies and the long duration of the intended treatments. Most juvenile animal studies were in the therapeutic areas of oncology, infectious diseases and endocrinology. In about 14% of the PIPs submitted, the NcWG requested either justifications of, or amendments to the study designs proposed by the applicants (e.g. justification of endpoints, study duration, species selection and timing with regards to clinical pediatric studies). Generally, only one species was selected or proposed for the juvenile studies, the rat being the most prevalent. The number of juvenile studies initially proposed by the applicant plus those requested by the NcWG was higher than the number of studies included in the "key binding elements" of the PIP opinions. This apparent discrepancy was mainly due to additional information or justifications submitted by the applicant during the clock stop. It was noted that the PIPs initially submitted often lacked information relevant to the nonclinical evaluation. Therefore, during the workshop, the need to provide scientifically based justifications when no juvenile animal studies are proposed in the initial PIP submission was stressed. PMID- 21638755 TI - Iron chelation treatment with deferasirox prior to high-dose chemotherapy and autologous stem cell transplantation may reduce the risk of hepatic veno occlusive disease in children with high-risk solid tumors. AB - BACKGROUND: We evaluated whether iron chelation treatment during induction chemotherapy could safely reduce serum iron levels and thereby reduce the frequency of hepatic veno-occlusive disease (VOD) during high-dose chemotherapy and autologous stem cell transplantation (HDCT/autoSCT) in children with high risk solid tumors. PROCEDURE: Children diagnosed with high-risk solid tumors between August 2008 and July 2009 were enrolled. Deferasirox treatment (25 mg/kg/day) was initiated when serum ferritin levels increased to more than 1,000 ng/ml during induction chemotherapy. Patients who were diagnosed with the same disease between April 2005 and June 2007 and treated in the same way without any iron chelation treatment formed the control group. Efficacy and toxicity of deferasirox treatment were compared between the two groups. RESULTS: Eighteen of 20 patients enrolled received deferasirox treatment. Deferasirox treatment was completed as scheduled in 11 (61.1%) of them without dose reduction or discontinuation. The serum ferritin levels prior to HDCT/autoSCT were lower in the deferasirox group than in the control group (median 1,268 ng/ml vs. 1,828 ng/ml, P < 0.001), although there was no difference in the RBC transfusion amount between the two groups. While 7 (17.9%) VODs developed during 39 HDCT/autoSCTs in the control group, there was no VOD during 40 HDCT/autoSCTs in the deferasirox group (P = 0.005). However, renal dysfunction (38.9%) including Fanconi syndrome (16.7%) was a frequently observed adverse effect of deferasirox treatment. CONCLUSIONS: Deferasirox treatment during induction chemotherapy reduces the frequency of VOD during HDCT/autoSCT. The development of renal dysfunction should be closely monitored during deferasirox treatment. PMID- 21638756 TI - The American Academy of Pediatrics policy statement--children as hematopoietic stem cell donors. PMID- 21638757 TI - Central nervous system late effects: a new frontier? PMID- 21638758 TI - CNS germinoma: one more piece of the puzzle. PMID- 21638759 TI - Sleep abnormalities in untreated patients with mucopolysaccharidosis type VI. AB - Mucopolysaccharidosis type VI (MPS VI) is a lysosomal storage disease that affects an enzyme responsible for the degradation of glycosaminoglycans (GAGs). Partially degraded GAGs accumulate in several tissues, such as the upper airways (UA), which leads to the development of obstructive sleep apnea (OSA). Our objective was to determine the prevalence of OSA in a group of untreated patients with MPS VI and the association of OSA with clinical and echocardiographic findings. Patients aged 4 years or older with a biochemical diagnosis of MPS VI were included. Data about clinical history, physical examination, Doppler echocardiogram, and overnight polysomnography (PSG) were collected. Our results showed that of the 28 participants, 14 were boys; mean age was 98.5 months, and mean age at MPS VI diagnosis was 48.4 months. Snoring, witnessed apnea, pectus carinatum, and macroglossia were the main clinical findings. PSG results showed that 23:27 patients (85.1%) had OSA which was mild in 4, moderate in 5, and severe in 14 patients. Echocardiograms showed evidence of pulmonary hypertension (PH) in 14 patients. Lower (P = 0.037) and nadir SpO(2) (P = 0.007) were positively associated with PH. Clinical signs suggestive of respiratory abnormalities during sleep were not significantly correlated with the results of PSG. We conclude that the prevalence of OSA in patients with MPS VI was high, and the level of desaturation was positively correlated with PH. Symptoms during sleep were not associated with PSG findings, which suggests that this population should undergo routine PSG as earlier as possible. This study provides baseline data to estimate the potential impact of specific treatments in the sleep abnormalities presented by patients with MPS VI. PMID- 21638760 TI - The role of cytomegalovirus in schizencephaly. PMID- 21638761 TI - Recurrent partial rhombencephalosynapsis and holoprosencephaly in siblings with a mutation of ZIC2. AB - Rhombencephalosynapsis (RES) is a rare congenital brain malformation typically identified by magnetic resonance imaging and characterized by fusion of the cerebellar hemispheres and dentate nuclei and vermian agenesis or hypogenesis. Although RES is frequently found in conjunction with other brain malformations and/or congenital anomalies, no specific molecular etiology has been discovered to date and no animal models exist. We identified two half sisters with alobar or semi-lobar holoprosencephaly (HPE) and partial RES, suggesting that genes linked to HPE may also contribute to RES. A deletion of seven base pairs in exon one of the ZIC2 gene (c.392_98del7) was identified in each of the two half sisters with HPE and partial RES. To identify genetic causes of RES and to assess whether genes identified in HPE have a role in RES, we tested 11 additional individuals with RES by high-resolution chromosome analysis, chromosomal microarray analysis, and sequencing of four HPE genes. No mutations in ZIC2 or in other genes that cause HPE were identified, suggesting that mutation of ZIC2 is a rare cause of, or contributor to, RES associated with HPE. In addition, an individual with a complex rearrangement of chromosome 22q13.3 and RES was identified, suggesting the presence of a dosage-sensitive gene that may contribute to RES in this region. PMID- 21638762 TI - Neurofibromatosis 2: a novel risk factor for hypertension? AB - There are anecdotal reports of neurofibromatosis 2 (NF2) patients and vascular disease, but no previous studies have compared blood pressure (BP) in people with NF2 and in the general population. This study is the first to show that BP is significantly higher in patients with NF2 than in matched control patients. This is important for the management of patients with NF2, and in the differential diagnosis of secondary hypertension. PMID- 21638763 TI - Genetic contribution for non-syndromic cleft lip with or without cleft palate (NS CL/P) in different regions of Brazil and implications for association studies. AB - Non-syndromic cleft lip with or without cleft palate (NS CL/P) is a complex disease in which heritability estimates vary widely depending on the population studied. To evaluate the importance of genetic contribution to NS CL/P in the Brazilian population, we conducted a study with 1,042 families from five different locations (Santarem, Fortaleza, Barbalha, Maceio, and Rio de Janeiro). We also evaluated the role of consanguinity and ethnic background. The proportion of familial cases varied significantly across locations, with the highest values found in Santarem (44%) and the lowest in Maceio (23%). Heritability estimates showed a higher genetic contribution to NS CL/P in Barbalha (85%), followed by Santarem (71%), Rio de Janeiro (70%), Fortaleza (64%), and Maceio (45%). Ancestry was not correlated with the occurrence of NS CL/P or with the variability in heritability. Only in Rio de Janeiro was the coefficient of inbreeding significantly larger in NS CL/P families than in the local population. Recurrence risk for the total sample was approximately 1.5-1.6%, varying according to the location studied (0.6-0.7% in Maceio to 2.2-2.8% in Barbalha). Our findings show that the degree of genetic contribution to NS CL/P varies according to the geographic region studied, and this difference cannot be attributed to consanguinity or ancestry. These findings suggest that Barbalha is a promising region for genetic studies. The data presented here will be useful in interpreting results from molecular analyses and show that care must be taken when pooling samples from different populations for association studies. PMID- 21638764 TI - A flow cytometric method to assess nanoparticle uptake in bacteria. AB - Toxicity of engineered nanomaterials (ENMs), such as metal oxides, has been of concern among environmental and health scientists. For ecotoxicity studies of ENMs, it is important to assess nanoparticle uptake and correlate it with the cellular response. However, due to nonavailability of adequate methods for assessing cellular uptake of ENMs, there is a lack of information in this important area. In the present study, a method has been developed using flow cytometry, which allows for rapid detection of ENM internalization in live bacteria under different experimental conditions for several generations. Our data demonstrate significant internalization of Zinc oxide (ZnO) and Titanium (IV) oxide (TiO(2) ) nanoparticles (NPs) in Escherichia coli in a dose-dependent manner. ZnO NPs treatment exhibited a significant increase in the intensity of side scatter (SSC) with liver-S9 fraction (76, 94, and 181% increase) rather than without S9 (10.5, 24.5, and 125.9% increase) at 10, 40, and 80 MUg/ml concentrations, respectively. This was due to the protein coating of NPs by the S9 fraction. A similar response was also observed on exposure to TiO(2) NPs (139 and 203% with S9 and 128 and 198% without S9). In a multigeneration study, this new method was able to detect the presence of ENMs in E. coli up to four generations. Our data demonstrate that this method can be used for assessing the uptake of ENMs in bacteria and provides a handle to toxicologists for ecotoxicity studies of economically important ENMs to ensure safer products in the market. PMID- 21638765 TI - Confocal backscattering-based detection of leukemic cells in flowing blood samples. AB - The prognostic value of assessing minimal residual disease (MRD) in leukemia has been established with advancements in flow cytometry and PCR. Nonetheless, these techniques are limited by high equipment costs, complex, and costly cell processing and the need for highly trained personnel. Here, we demonstrate the potential of exploiting differences in the relative intensities of backscattered light at three wavelengths to detect the presence of leukemic cells in samples containing varying mixtures of white blood cells (WBCs) and leukemic cells flowing through microfluidic channels. Using 405, 488, and 633 nm illumination, we identify distinct light scattering intensity distributions for Nalm-6 leukemic cells, normal mononuclear (PBMC) and polymorphonuclear (PMN) white blood cells and red blood cells. We exploit these differences to develop cell classification algorithms, whose performance is evaluated based on simultaneous acquisition of light scattering and fluorescence flow cytometry data. When this algorithm is used prospectively for the analysis of samples consisting of mixtures of PBMCs and leukemic cells, we achieve an average specificity and sensitivity of leukemic cell detection of 99.6 and 45.2%, respectively. When we consider samples that include leukemic cells along with PMNs and PBMCs, which can be acquired using a simple red blood cell lysis step following venipuncture, the specificity and sensitivity of the approach decreases to 91.6 and 39.5%, respectively. On the basis of the performance of these algorithms, we estimate that 42 or 71 MUL of blood would be adequate to confirm the presence of leukemia at an 80% power level in samples containing 0.01% leukemia to either PBMCs or PBMCs and PMNs, respectively. Therefore, light scattering-based flow cytometry in a microfluidic platform could provide a low cost, highly portable, minimally invasive approach for detection and monitoring of leukemic patients. This could offer significant improvements especially for pediatric patients and for patients in developing countries. PMID- 21638766 TI - An imaging flow cytometric method for measuring cell division history and molecular symmetry during mitosis. AB - Asymmetric cell division is an important mechanism for generating cellular diversity, however, techniques for measuring the distribution of fate-regulating molecules during mitosis have been hampered by a lack of objectivity, quantitation, and statistical robustness. Here we describe a novel imaging flow cytometric approach that is able to report a cells proliferative history and cell cycle position using dye dilution, pH3, and PI staining to then measure the spatial distribution of fluorescent signals during mitosis using CCD-derived imagery. Using Jurkat cells, resolution of the fluorescently labeled populations was comparable to traditional PMT based cytometers thus eliminating the need to sort cells with specific division histories for microscopy. Subdividing mitotic stages by morphology allowed us to determine the time spent in each cell cycle phase using mathematical modeling approaches. Furthermore high sample throughput allowed us to collect statistically relevant numbers of cells without the need to use blocking agents that artificially enrich for mitotic events. The fluorescent imagery was used to measure PKCzeta protein and EEA-1+ endosome distribution during different mitotic phases in Jurkat cells. While telophase cells represented the favorable population for measuring asymmetry, asynchronously dividing cells spent approximately 43 seconds in this stage, explaining why they were present at such low frequencies. This necessitated the acquisition of large cell numbers. Interestingly we found that PKCzeta was inherited asymmetrically in 2.5% of all telophasic events whereas endosome inheritance was significantly more symmetrical. Furthermore, molecular polarity at early mitotic phases was a poor indicator of asymmetry during telophase highlighting that, though rare, telophasic events represented the best candidates for asymmetry studies. In summary, this technique combines the spatial information afforded by fluorescence microscopy with the statistical wealth and objectivity of traditional flow cytometry, overcoming the key limitations of existing approaches for studying asymmetry during mitosis. PMID- 21638768 TI - Case Study Interpretation--Houston: Case 1. Phenytoin-induced ''pseudolymphoma''. PMID- 21638769 TI - Case Study Interpretation--Houston: Case 2. Classical Hodgkin lymphoma. PMID- 21638770 TI - Case Study Interpretation--Houston: Case 3. Early T-cell precursor-acute lymphoblastic leukemia. PMID- 21638771 TI - Case Study Interpretation--Houston: Case 4. T-cell prolymphocytic leukemia. PMID- 21638772 TI - No effect of lysis solutions on absolute CD19+ lymphocytes count and CD45 index in chronic lymphocytic leukemia. AB - The absolute CD19+ lymphocytes count is essential for chronic lymphocytic leukemia (CLL) management. At the present time, no standardized flow cytometry (FCM) protocol to measure B-lymphocytes counts is established. The aims of the present study were first to evaluate the effect of different lysis solutions and of red blood cell lysis per se on CLL lymphocytes count and B-lymphocytes CD45 expression and second to compare absolute B-lymphocytes counts obtained by single (SP) and dual platforms (DP). Absolute CD19+ B-lymphocytes counts and CD45 expression in 35 whole-blood CLL samples were determined by FCM using either different lysis solutions or using a no wash no lyse (NWNL) protocol. Single platform using microbeads was also evaluated for absolute quantification. The absolute CD19+ B-lymphocytes counts using different red blood cell lysis solutions correlated with NWNL method without any effect on CD45 expression. Bland and Altman plot showed homogenous distribution of bias; mean bias was less than 1% for all lysing solutions. Moreover, no statistically significant difference between SP and DP was observed. The type of lysis solution influences neither the CD19+ B-lymphocytes count nor the CD45 expression. The two systems, SP and DP, yield comparable values with excellent agreement. However, the tendency of slightly lower results with SP showed the requirement of larger studies before standardization of B-lymphocytes count in CLL patients. PMID- 21638775 TI - Primary prevention of cancer is a global priority. PMID- 21638773 TI - Cell-cycle distribution of different cell compartments in normal versus reactive bone marrow: a frame of reference for the study of dysplastic hematopoiesis. AB - Limited information is currently available about the proliferation activity and cell-cycle distribution of different bone marrow (BM) cell subsets defined according to their lineage and maturation stage in normal versus cytopenia associated reactive BM samples. Here, we report a three-color flow cytometry approach to investigate the cell-cycle distribution of different BM cell compartments-CD34(+) hematopoietic progenitor and precursor cells (HPC), maturing neutrophils and monocytic cells, mature lymphocytes, eosinophils, and nucleated red blood cell precursors (NRBC)-from normal (n = 47) versus cytopenia-associated reactive (n = 47) BM samples. Highly similar proliferation profiles were detected in normal versus reactive BM, with a higher proliferation index (PI) for the more immature CD34(+) HPC, CD11b(-) maturing neutrophils and NRBC versus other BM cell compartments. The only differences observed between normal and reactive BM were restricted to the more mature (CD13(hi) /CD11b(+) ) bands/neutrophils and to monocytic cells, which showed an increased PI (0.9% +/- 0.8% vs. 0.6% +/- 0.5% and 6 +/- 3.6 vs. 4.6 +/- 4.5, respectively) at the expense of a lower PI of CD34(+) HPC in reactive conditions. Of note, bands/mature neutrophils and mature lymphocytes showed either residual numbers or absence of S + G2 /M-phase cells in both normal and reactive BM. Our results suggest that a slight shift of proliferation from the early precursors to the more mature granulomonocytic compartment occurs in reactive BM, which could reflect an attempt of the hematopoietic system to rapidly produce functional neutrophils and monocytes, at the expense of a lower expansion of the minor compartments of CD34(+) HPC. PMID- 21638776 TI - Release of lipophilic molecules during in vitro digestion of soy protein stabilized emulsions. AB - SCOPE: Solubilization of lipophilic bioactives in gastrointestinal fluids contributes to their bioavailability, but a better understanding of the transfer processes involved and the impact of molecular structure is required. METHODS AND RESULTS: The transfer of beta-carotene (BC), coenzyme Q10 (CoQ10), vitamin D3 (VitD3), and phytosterols (PSs) from soy protein isolate-stabilized oil-in-water emulsions to the aqueous phase during in vitro digestion was investigated. In the absence of lipolysis, transfer was mainly governed by molecular structure and partitioning within the oil droplets. Less than 3% BC and CoQ10, versus 30.4 +/- 0.3% PSs and 24.7 +/- 0.4% VitD3, were transferred in this case. However, with lipolysis, PSs and VitD3 rapidly partitioned into the aqueous phase, while lag phases and slower transfer rates were observed for BC and CoQ10. Positive and linear correlations between lipolysis and transfer were observed for all systems. After 2 h exposure to simulated duodenal conditions, there were no differences between percent micellization, except for BC which was proportionally lower. VitD3 and PSs mutually enhanced each other's transfer, while no interactions were observed between VitD3 and BC. CONCLUSION: Bioactive molecular structure and co administration influenced the transfer behaviour, with implications for foods designed to optimize health benefits. PMID- 21638777 TI - Bacterial biofilms associated with food particles in the human large bowel. AB - Bacteria within the gastro-intestinal tract affect host function via production of short-chain fatty acids and synthesis of vitamins. Additionally, the commensal enteric bacteria modulate the immune system and provide protection from potentially pathogenic bacteria. Only recently heterogeneous bacterial biofilms were found to be associated with food particles within the intestinal tract. There are a number of studies investigating the formation and function of pathogenic and single-species biofilms, though few studies have investigated the dynamics of multispecies biofilms, especially with regard to food/microbial/host interactions. The scope of this review is to discuss the current knowledge of bacterial biofilms associated with food particles in the human large bowel, examine the established mathematical models depicting bacterial attachment, and elucidate key areas for further research. PMID- 21638778 TI - Dietary-resistant starch improves maternal glycemic control in Goto-Kakizaki rat. AB - SCOPE: Dietary prebiotics show potential in anti-diabetes. Dietary resistant starch (RS) has a favorable impact on gut hormone profiles, including glucagon like peptide-1 (GLP-1) consistently released, a potent anti-diabetic incretin. Also RS reduced body fat and improved glucose tolerance in rats and mice. In the current project, we hypothesize that dietary-resistant starch can improve insulin sensitivity and pancreatic beta cell mass in a type 2 diabetic rat model. Altered gut fermentation and microbiota are the initial mechanisms, and enhancement in serum GLP-1 is the secondary mechanism. METHODS AND RESULTS: In this study, GK rats were fed an RS diet with 30% RS and an energy control diet. After 10 wk, these rats were mated and went through pregnancy and lactation. At the end of the study, pancreatic beta cell mass, insulin sensitivity, pancreatic insulin content, total GLP-1 levels, cecal short-chain fatty acid concentrations and butyrate producing bacteria in cecal contents were greatly improved by RS feeding. The offspring of RS-fed dams showed improved fasting glucose levels and normal growth curves. CONCLUSION: Dietary RS is potentially of great therapeutic importance in the treatment of diabetes and improvement in outcomes of pregnancy complicated by diabetes. PMID- 21638779 TI - Response of a Lactobacillus plantarum human isolate to tannic acid challenge assessed by proteomic analyses. AB - SCOPE: To gain insight on the mechanisms used by intestinal bacteria to adapt and resist the antimicrobial action of dietary tannins and identify targets for tannic acid in Lactobacillus plantarum. METHODS AND RESULTS: A proteomic analysis of an L. plantarum human isolate exposed to the tannic acid challenge was undertaken. Tannic acid targeted proteins involved in outstanding processes for bacterial stress resistance including cyclopropanation of membrane lipids, stress response at population scale and maintenance of cell shape. To respond to this aggression, tannic acid-misfit cells of L. plantarum challenged with tannic acid reorganized their metabolic capacity to economize energy and express proteins involved in oxidative stress defense and cell wall biogenesis, indicating that the injury incurred by tannic acid was based on oxidative damage and disruption of the cell envelope. The induction of 3-octaprenyl-4-hydroxybenzoate carboxy lyase, which is sensitive to changes in redox conditions and involved in ubiquinone biosynthesis in other bacteria, suggests for a tannic acid-induced redox imbalance. CONCLUSION: The results reveal the adaptation of a gastrointestinal isolate of L. plantarum to tannic acid and identify antibacterial targets for this dietary compound. This provides the basis for the selection of tannin-resistant microorganisms and their use to obtain health benefits from tannin-containing diets. PMID- 21638780 TI - Chemical preparation of graphene-based nanomaterials and their applications in chemical and biological sensors. AB - Graphene is a flat monolayer of carbon atoms packed tightly into a 2D honeycomb lattice that shows many intriguing properties meeting the key requirements for the implementation of highly excellent sensors, and all kinds of proof-of-concept sensors have been devised. To realize the potential sensor applications, the key is to synthesize graphene in a controlled way to achieve enhanced solution processing capabilities, and at the same time to maintain or even improve the intrinsic properties of graphene. Several production techniques for graphene based nanomaterials have been developed, ranging from the mechanical cleavage and chemical exfoliation of high-quality graphene to direct growth onto different substrates and the chemical routes using graphite oxide as a precusor to the newly developed bottom-up approach at the molecular level. The current review critically explores the recent progress on the chemical preparation of graphene based nanomaterials and their applications in sensors. PMID- 21638781 TI - Enhanced electrical conductivity of individual conducting polymer nanobelts. PMID- 21638782 TI - A light trigger for DNA nanotechnology. PMID- 21638784 TI - Self-organization of thin polymer films guided by electrostatic charges on the substrate. AB - The self-organization of thin polymer films into functional patterns is important both scientifically and technologically. Electric fields have been exploited as an efficient and powerful means to induce the destabilization and self organization of soft materials. Previous attention, however, has mainly focused on externally applied electric fields. It is shown herein that the internal electric field is strong enough to guide the self-organization of thin polymer films as well. Patterns of electrostatic charges with micrometer resolution are first introduced on a dielectric substrate. A thin polymer film is then spin coated onto the topographically flat substrate. Upon thermal annealing, the thin polymer film destabilizes due to a lateral gradient of electrostatic stress and flows away from the electroneutral regime to the charged area, resembling the patterns of charges on the substrate. Theoretical and numerical modeling based on the electrohydrodynamic instability shows excellent agreement with experimental observations both qualitatively and quantitatively. It is also demonstrated that the interplay between charge-driven instability with spinodal dewetting and Rayleigh instabilities can generate finer and hierarchical polymeric patterns that are completely different from the charge patterns preintroduced on the substrate. This study provides direct evidence that the internal electric field caused by charges on the substrate is strong enough to destabilize thin polymeric films and generate patterns. This study also demonstrates new strategies for bottom-up fabrication of structured functional materials. PMID- 21638783 TI - Molecular analysis of blood with micro-/nanoscale field-effect-transistor biosensors. AB - Rapid and accurate molecular blood analysis is essential for disease diagnosis and management. Field-effect transistor (FET) biosensors are a type of device that promise to advance blood point-of-care testing by offering desirable characteristics such as portability, high sensitivity, brief detection time, low manufacturing cost, multiplexing, and label-free detection. By controlling device parameters, desired FET biosensor performance is obtained. This review focuses on the effects of sensing environment, micro-/nanoscale device structure, operation mode, and surface functionalization on device performance and long-term stability. PMID- 21638785 TI - The passage of homopolymeric RNA through small solid-state nanopores. AB - Solid-state nanopores are widely acknowledged as tools with which to study local structure in biological molecules. Individual molecules are forced through a nanopore, causing a characteristic change in an ionic current that depends on the molecules' local diameter and charge distribution. Here, the translocation measurements of long (~5-30 kilobases) single-stranded poly(U) and poly(A) molecules through nanopores ranging from 1.5 to 8 nm in diameter are presented. Individual molecules are found to be able to cause multiple levels of conductance blockade upon traversing the pore. By analyzing these conductance blockades and their relative incidence as a function of nanopore diameter, it is concluded that the smallest conductance blockades likely correspond to molecules that translocate through the pore in predominantly head-to-tail fashion. The larger conductance blockades are likely caused by molecules that arrive at the nanopore entrance with many strands simultaneously. These measurements constitute the first demonstration that single-stranded RNA can be captured in solid-state nanopores that are smaller than the diameter of double-stranded RNA. These results further the understanding of the conductance blockades caused by nucleic acids in solid-state nanopores, relevant for future applications, such as the direct determination of RNA secondary structure. PMID- 21638786 TI - Mixed-ligand nanoparticles as supramolecular receptors. PMID- 21638787 TI - Size-controlled peptide-directed synthesis of hollow spherical gold nanoparticle superstructures. PMID- 21638789 TI - [Why not protect the "liquidators" at Fukushima?]. PMID- 21638788 TI - Self-assembly and applications of nucleic acid solid-state films. AB - While most nucleic acid (NA)-lipid or NA-polymer complexes are studied in solution, there is growing interest in understanding their properties as naturally derived, biodegradable, biocompatible, solid-state materials with tailorable properties influenced by environmental parameters. Therapeutic and cell programming applications comprise an important new research field, particularly in gene transfection and silencing using plasmid DNA and siRNA with targeted local delivery for use in cell culture. Dried solid films have lower nuclease degradation, fewer barriers to long term storage, and allow localized delivery by direct implantation in combination with controlled release and dosage adjustment. In contrast to particulate complexes or other methods of drug delivery which are prepared and must remain in solution, films can regain their biological activity once wetted. However our understanding of the types of cationic agents that predictably form self-standing films with NA is still limited. The self-assembly and structural, physical, and chemical properties of these materials are of key importance to maintaining their activity. We therefore discuss the material properties of NA-lipid and NA-polymer films as the focus of this article. Recent studies have indicated that there is also growing interest in NA films beyond bioengineering and medical applications in the fields of nano- and optoelectronics. We survey the self-assembly of solid-state materials composed of NA complexed with lipids, surfactants, or polymers, and summarize investigations of nanoscopic assembly, structure, optical, and macroscopic material properties. We further evaluate the current and future applications of NA-lipid and NA-polymer films and the benefits and drawbacks of each type. PMID- 21638790 TI - Abstracts of the 14th International Congress of ESCAP European Society for Child and Adolescent Psychiatry. June 11-15, 2011. Helsinki, Finland. PMID- 21638791 TI - [Proceedings from the Congress of the French Society of Pediatrics, 11-14 May 2011, Marseille, France]. PMID- 21638792 TI - Abstracts of the 2011 Vascular Annual Meeting. Chicago, Illinois. USA. June 16 18, 2011. PMID- 21638793 TI - Strigolactones are regulators of root development. AB - Strigolactones (SLs) have been defined as a new group of plant hormones or their derivatives that suppress lateral shoot branching. Recently, a new role for SLs was discovered, in the regulation of root development. Strigolactones were shown to alter root architecture and affect root-hair elongation. Here, I review the recent findings regarding the effects of SLs on root growth and development, and their association with changes in auxin flux. The networking between SLs and other plant hormones that regulate root development is also presented. Strigolactone regulation of plant development suggests that they are coordinators of shoot and root development and mediators of plant responses to environmental conditions. PMID- 21638794 TI - Halting the rise of non-communicable diseases: an urgent priority for women's health. PMID- 21638795 TI - Abstracts of the American Society of Gene & Cell Therapy 14th Annual Meeting. May 18-May 21, 2011. Seattle, Washington, USA. PMID- 21638796 TI - Abstracts of CARS 2011: Computer Assisted Radiology and Surgery. Proceedings of the 25th International Congress and Exhibition. Berlin, Germany. June 22 - 25, 2011. PMID- 21638797 TI - Plant growth and architectural modelling and its applications. Preface. AB - Over the last decade, a growing number of scientists around the world have invested in research on plant growth and architectural modelling and applications (often abbreviated to plant modelling and applications, PMA). By combining physical and biological processes, spatially explicit models have shown their ability to help in understanding plant-environment interactions. This Special Issue on plant growth modelling presents new information within this topic, which are summarized in this preface. Research results for a variety of plant species growing in the field, in greenhouses and in natural environments are presented. Various models and simulation platforms are developed in this field of research, opening new features to a wider community of researchers and end users. New modelling technologies relating to the structure and function of plant shoots and root systems are explored from the cellular to the whole-plant and plant community levels. PMID- 21638798 TI - Women at risk: why increasing numbers of women are failing to get the health care they need and how the Affordable Care Act will help. Findings from the Commonwealth Fund Biennial Health Insurance Survey of 2010. AB - Women have greater health care needs than men, and generally play larger roles in the health care of family members. Rising health care costs combined with sluggish income growth has contributed to losses in health insurance among women and rising rates of problems gaining necessary health care and paying medical bills. Women who seek coverage in the individual insurance market face additional hurdles--few plans offer maternity coverage and, in most states, insurance carriers charge higher premium rates to young women than men of the same age. The Affordable Care Act is bringing change for women through required free coverage of preventive care services, small business tax credits, new affordable coverage options, and insurance market reforms, including bans on gender rating. When the law is fully implemented in 2014, nearly all the 27 million working-age women who went without health insurance in 2010 will gain affordable and comprehensive benefits. PMID- 21638799 TI - Maintaining coverage, affordability, and shared responsibility when income and employment change. AB - The Affordable Care Act builds on existing sources of public and private health insurance, while creating new health insurance exchanges and subsidies. A potential disadvantage of preserving many sources of health insurance is the likelihood of abrupt changes in coverage or financial responsibility when individual circumstances change. This brief describes four policy challenges related to such changes: adjusting premium and cost-sharing subsidies when incomes change; coordinating eligibility for premium credits, Medicaid, and the Children's Health Insurance Program; encouraging and facilitating continuous coverage; and minimizing transitions between individual and small-business exchanges. Policy recommendations to reduce uncertainty, simplify coverage decisions, and minimize insurance transitions include extending coverage to the open enrollment period at the end of the year, generous treatment of income gains in correcting premium tax credits, and unifying the small-business and individual exchanges. PMID- 21638800 TI - How the Affordable Care Act is helping young adults stay covered. AB - The Affordable Care Act is making a difference for young adults, among the groups most at risk for lacking health insurance in the United States. Young adults up to age 26 may now stay on or join their parent's health plans if they include dependent coverage, and early reports indicate that at least 600,000 have done so. Starting in 2014, of the 14.8 million uninsured adults ages 19 to 29, an estimated 12.1 million could gain subsidized coverage once all the law's provisions go into effect: 7.2 million may gain coverage under Medicaid and 4.9 million may gain subsidized private coverage through state insurance exchanges. New findings from the 2010 Commonwealth Fund Biennial Health Insurance Survey indicate the need for these reforms: 45 percent of young adults reported delaying needed care because of costs in 2010, up from 32 percent in 2001, and 39 percent reported problems paying medical bills. PMID- 21638801 TI - Review of mastitis diagnoses in Scottish cattle in 2010. PMID- 21638802 TI - [Species-area relationship at different succession stages of monsoon evergreen broad-leaved forest in south subtropical area of Yunnan Province]. AB - Based on the investigation data of monsoon evergreen broad-leaved forest at its different succession stages (primary, CP; 15 years of succession, CF; and 30 years of succession, CT) in Pu' er of Yunnan Province, this paper studied the species-area relationship of this forest at each succession stage. It was found that in the communities at each succession stage, the number of total species, trees, shrubs, and lianas had a significant correlation with sampling area, with the area explained over 94% of the total variation. The Z value of the total species (0.334) and trees (0.394) was the lowest at CT, whereas that of shrubs (0.437) and lianas (0.326) was the lowest at CF. No significant differences were observed in the intercepts of the species-area curve of total species, trees, shrubs, and lianas among different succession stages, but the coefficient of determination (R2) of the species-area curve of total species and lianas was the highest at CP. The richness of trees and shrubs at CF explained 99.9% of the variance of Z value, but the richness of total species, trees, shrubs, and lianas at CP and CT had no significant correlations with the Z value. PMID- 21638803 TI - [25 years of organized ambulatory heart sport in Luxembourg. The development of a sustained rehabilitation model]. AB - After Second World War cardiovascular disease (CVD), especially coronary artery disease (CAD), turned out to be an epidemic in the western countries including the Grand-Duchy of Luxembourg, and accounted for nearly half of all deaths. A lot of strategies, among them treatment by physical activity, were developed to fight this challenge and, fortunately, a mortality regression of about 50 % could be achieved. Nowadays, eastern European countries and, more recently, China and India face an increasing CVD mortality. During the seventies ambulatory heart sport clubs, then labeled as, "coronary clubs" became very popular in Europe especially in West-Germany. Around 2000 there were more than 6000 heart sports groups in Germany. In 1984 a first group was founded in Luxembourg City (Centre) a, 1991 a second group started in Esch/Alzette (South) and in 2002 a third regional group in Ettelbruck (North) so that, by now, the 3 main public health districts of the Grand-Duchy of Luxembourg can offer regular ambulatory sports activities to almost all concerned cardiac patients in the country. The ambulatory heart sport groups of Luxembourg cooperate in a federated association allowing an integrated logistic organization. Since the beginning nineties cardiac rehabilitation became a field of interest to the university faculties and later of scientific societies, like the American Heart Association (AHA) and the European Society of Cardiology (ESC). New subgroups were founded and guidelines were published and renewed. The movement of ambulatory heart sport groups was more or less neglected in the prevention and rehabilitation activities of the scientific societies. Recently the ESC proposed a new definition of comprehensive CVD prevention and rehabilitation programmes as "coordinated, multifaceted interventions designed to optimize a cardiac patient's physical, psychological, and social functioning, in addition to stabilizing, slowing, or even reversing the progression of the underlying atherosclerotic processes, thereby reducing morbidity and mortality". The responsible ESC cardiologists agree with the international community that fighting CVD risk factors is at least as important as the whole arsenal of modern heart surgery and interventional cardiology. The core activity of ambulatory heart sport groups remains physical activity, and nowadays 6 different activities can be offered (one activity each day of the week): exercise lesson, swimming, walking, cycling, Nordic Walking and water gymnastics On the other hand comprehensive prevention programs, especially concerning CVD risk factors are also endorsed by the ambulatory heart sport groups of Luxembourg via regular meetings, conferences, brochures and symposia. One advantage of the ambulatory heart sport movement in Luxembourg, in contrast to the German model, is the direct financial allowance of the health ministry, which permits a lifelong activity to all the active members. Another advantage is that all the regional groups are directed by clinical cardiologists knowing the patients very closely. One weak point is that only about 5-10% of all potential candidates adhere to the ambulatory heart sport groups but nearly 50% of the active members are practicing for more than 5 years. These regularly active patients are a positive selection of well committed cardiac patients who, most of the time, control CVD risk factors with scrutiny. The ESC has recommended creating so called "Heart Houses" where all the aspects of comprehensive prevention and rehabilitation can be offered. Their main concern is to develop a sustained strategy which is desperately missing for the moment. A lot of the active members of the heart sport groups of Luxembourg achieve such a sustained activity and, therefore, these heart sport groups can be considered as very cost effective models of sustained rehabilitation. After a 25 years activity the ambulatory heart sport movement of Luxemburg has reached the outstanding goal of being a center of sustained rehabilitation. Although such a goal was not really planned during the first 15 years of activity, the stamina of the active members set the movement into this direction. PMID- 21638804 TI - [Diversity of blood parasites of genus Bartonella in wild rodents in Mazury Lakes District]. AB - This long-term study of genetic diversity and epidemiology of the alpha proteobacterium Bartonella in wild rodents from forest (Myodes glareolus and Apodemus flavicollis) and abandoned farmland (Microtus arvalis and Mi. oeconomus) was carried out in the years 2007-2009 in the Mazury Lakes District. In total, 1193 rodents were marked and recaptured, and 2226 blood samples were collected. The highest Bartonella prevalence was found in A. flavicollis (43.5%), the lowest in Mi. oeconomus (9.4%), while prevalence in My. glareolus and Mi. arvalis was, respectively, 13.2% and 11.8% (PCR of citrate synthase gltA gene fragment). Prevalence varied according to year and season, as well as sex of rodents. For woodland animals, a rapid decrease of prevalence was observed in late 2008, due to the dilution effect. Multiple (different species/genotypes of Bartonella in successive months) and mixed infections (more than one bacteria genotype in the same blood sample) were also diagnosed. Between 2835 and 4800000 colony forming units (CFU) per ml blood were recorded, with, for B. taylorii, significant differences between isolates from hosts belonging to different host families. Sequence analysis of 147 isolates revealed 37 gltA variants. In all four rodents, B. taylorii was the most prevalent, and could be divided into three main clades. One clade of B. grahamii was present in My. glareolus, A. flavicollis and Mi. arvalis, and both Microtus species were infected with a single clade of B. doshiae. A single isolate of B. birtlesii from A. flavicollis was collected, while two isolates could not be assigned to any known species. Nested clade analysis showed host specificity of 1st step clades (connected with rodent species) and 2nd step clades (connected with rodent family). Analysis was then extended to other housekeeping genes (cell division proteinftsZ, heat shock protein groEl, riboflavin synthase ribC, beta subunit RNA polymerase rpoB) and gene encoding 16S rRNA. Comparison of alleles of these genes in 27 isolates revealed numerous recombinant events, primarily involving groEl and 16S rRNA genes. Moreover, genetic recombination within housekeeping genes was also identified, and one of the unidentified Bartonella isolates was found to involve recombination within gltA between B. grahamii and B. taylorii. Examination of two T4SS pathogenicity genes (virB5 and bepA), revealed a similar pattern of extensive recombination. BepA from 17 isolates showed little diversity, concomitant with its role as an intra-cellular messenger. The virB5 gene (encoding a putative extra-cellular adhesin) from 22 isolates from voles (Arvicolidae) and A. flavicollis was distinctively different in sequence and putative structure, and showed a clear signature of horizontal gene transfer. Moreover, these recombinant events were often identified in the same isolates in which recombination of groEl or 16S rRNA was observed, suggesting that selection for this pathogenicity gene is important in the microevolution of Bartonella within rodents. In particular, Microtus spp. was central in the appearance of novel Bartonella isolates. PMID- 21638805 TI - [Educational conference "Environmental and molecular aspects of parasitism"]. PMID- 21638806 TI - [Protocol of the Polish Parasitology Society (PT)Congress on January 21, 2011]. PMID- 21638807 TI - [An unusual finding in thoracic CT]. PMID- 21638808 TI - [Regardless of the reason for her consultation, examine every child also orthopedically]. PMID- 21638809 TI - [Chronic cough and suspicious opacities in the roentgen image: pneumonia, lymphoma or metastases?]. PMID- 21638810 TI - [Etiology of increased liver values from alcohol to hemochromatosis. Many roads lead to cirrhosis]. PMID- 21638811 TI - [Specific general practitioner guidelines--why they deserve greater acceptance, dissemination and application]. PMID- 21638812 TI - [Management of the difficult airway in outpatient emergency--what if the tube cannot be placed?]. PMID- 21638813 TI - [Patellar dislocation]. PMID- 21638814 TI - [Emergency checklist: acute scrotum]. PMID- 21638815 TI - [Acute stroke in hypertensive patients: treat hypertension slowly and check blood pressure continuously]. PMID- 21638816 TI - [Differential diagnosis and therapy examined under the microscope. LUTS (lower urinary tract symptoms) due to OAB (overactive bladder) - a frequent disease picture in the man]. PMID- 21638817 TI - [Physiopathology and management of sickle cell in the adult]. PMID- 21638818 TI - [Management of the hospitalized patient with acute sickle cell crisis]. PMID- 21638819 TI - [Prevention of complications associated with acute sickle cell crisis]. PMID- 21638820 TI - [Dietary intervention and adaptation for the sickle cell patient]. PMID- 21638821 TI - [Skin ulcers and sickle cell]. PMID- 21638822 TI - [Social support and treatment planning of patients with sickle cell]. PMID- 21638823 TI - [Psychological support of the sickle cell patient]. PMID- 21638824 TI - [The role of the nurse in the diagnosis of cancer]. PMID- 21638825 TI - [Body, culture and health: the communication gap]. PMID- 21638826 TI - [Waist circumference, an important parameter in nursing practice]. PMID- 21638827 TI - [Important of nursing care in the administration of oral medications]. PMID- 21638828 TI - [The use of hypnosis by health professionals]. PMID- 21638829 TI - [Nursing training via apprenticeship contract]. PMID- 21638830 TI - [Orotracheal intubation]. PMID- 21638831 TI - [Schizophrenia]. PMID- 21638832 TI - [Big Sisters for supporting future mothers who are HIV seropositive]. PMID- 21638833 TI - [Image of the month. Occupational diseases of hairdressers: interdigital pilonidal sinus]. PMID- 21638834 TI - [Clinical case of the month. Portal venous gas due to a hydrogen peroxide colic enema]. AB - Portal venous gas is a worrying radiological sign that was described for the first time in 1955 among newborn children suffering from necrotizing enterocolitis. It was often related to mesenteric ischemia which mortality remains very high. Now, as radiological explorations increase, portal venous gas is more and more described in less pejorative contexts, such as hydrogen peroxide intoxication. It is what happened to our patient who underwent a hydrogen peroxide colic enema followed by a major and transient portal venous gas onset. This solved quickly and spontaneously. PMID- 21638835 TI - [Metabolic surgery: an evolution (or revolution) of bariatric surgery?]. AB - Bariatric surgery has proven its efficacy to obtain a marked and sustained weight loss and dramatically improves metabolic control in obese patients. The frequently observed remission of type 2 diabetes occurs very early, before any marked weight reduction. Increasing evidence suggests that this favourable effect results from profound changes in gut hormones involved in the regulation of energy intake behaviour and glucose homeostasis rather than simply from mechanical food restriction or malabsorption imposed by the surgical procedure. The better knowledge of these pathophysiological mechanisms, especially well studied with Roux-en Y gastric bypass, resulted in recent innovation in the technical procedures leading to a shift from bariatric surgery to metabolic surgery. Such type of surgery is currently evaluated in patients with type 2 diabetes, but with only a moderate obesity (BMI < 35 kg/m2), or even without obesity (BMI < 30 kg/m2). The Belgian Metabolic Intervention (BMI) Study Group would like to contribute very soon to this evaluation in a multidisciplinary approach. PMID- 21638836 TI - [Skin tanning and skin bleaching: ethnic addictions complying with the latest fad]. AB - Sun is at the origin of life and is responsible for death as well. In the past decades cutaneous melanoma has affected more and more young adults. In the White population, sun exposures and sunbeds are pointed out as the main culprits responsible for addictive behaviour. Preventive measures to be taken aim at avoiding the effects of ultraviolet light addiction. Sunlight exposure particularly in moderation is indeed beneficial through vitamin D3 synthesis and improved psychological wellbeing. Hence, we are facing the dilemma between too much and too little ultraviolet light exposure.In the Black population, the reverse situation is prevalent. It concerns skin depigmentation which may turn out to be a caricature in some subjects. PMID- 21638837 TI - [Antidepressants of the future: role of the glutamatergic system]. AB - Despite the availability of several antidepressants, the treatment of major depression is far from being satisfactory. With conventional antidepressants, more than 30 to 45% of the patient will not respond or present only partial remission. In the prospect of a better treatment of depression, research tends to develop original molecules, more effective, with a faster onset of action and a better tolerability. The new targets of antidepressive pharmacotherapy are beyond the membrane receptor. The future in the treatment of depression requires a better understanding of cellular and molecular mechanisms involved in the pathophysiology of depression and biochemical mechanisms explaining the antidepressive effect. In this context, the glutamatergic system plays a role in the pathophysiology of depression. Agents modulating glutamatergic activity could act as antidepressants. PMID- 21638838 TI - [Patient complaints in a hospital emergency department in Belgium]. AB - Patients express their dissatisfaction through complaints. This study analyzed the frequency and chief complaints of patients presenting to the emergency department (ED). The end point was find ways to improve patient satisfaction after their ED visit. In this retrospective, seven years study, we reviewed 155 chief complaints of patients presenting to the ED of a university hospital. The chief complaints were either from the patients or a family member. One hundred and fifty five chief complaints collected from 496.816 patients presenting to the ED were reviewed over a period of seven years. Complaints case rate was 3.1 per 10.000 visits. Complaints came from patients between the age of 20 to 60 years old (75.0 percent). Complains involved a physician (79.0 percent). The complaints were related a lack of communication (39.0 percent), long waiting time (14.0 percent), wrong diagnosis (22.0 percent), wrong treatment (13.0 percent) and ED disposition of the patient (12.0 percent). Two types of pathology represented more than 15 percent of the complaints: the traumatology (22.0 percent) and the psychiatry (17.0 percent). The traumatology and psychiatry represented respectively 30.0 percent and 10.0 percent of ED visits. Most complaints were addressed and resolved through a hospital mediator, Chief of service or Chief of staff. The rate of complains is low. Most complaints can be prevented if the physician improves communication with patients. PMID- 21638839 TI - [Prescribing practices of fluoroquinolones in Tunisia]. AB - The aim of the study was to assess the prescribing practices of fluoroquinolones (FQ) among general practionners in the town of Sousse (central-eastern Tunisia). A transversal study was made between the first and the 23rd April, 2009, among general practionners working in Sousse. For each prescription, informations about patients, indications and modalities of treatment were collected on a questionnaire. These prescriptions were compared to Tunisian and/or French guidelines for antibiotics use. One hundred and eighty eight FQ prescriptions were analyzed. The mean age of patients was 50 years. FQ were more often used alone (83%) and in first line intention (84%). The molecules used were essentially ciprofloxacin (44.7%), levofloxacin (35.6%) and ofloxacin (18.6%). The indications were mainly bronchopulmonary infections (34%) and urinary tract infection (32%). These choices were in accordance with guidelines in 41% of the prescriptions. The dosage was adapted, but the duration of treatment was often excessive. Further efforts are needed, to optimize the good use of FQ in order to reduce or stabilize the rate of bacterial resistance. PMID- 21638840 TI - [Medication of the month. Femoston Low (0.5 mg estradiol plus 2.5 mg dydrogesterone) for menopausal hormonal replacement therapy]. AB - Femoston Low is a hormone replacement therapy that combines low dosages of steroids, i.e. 0.5 mg of estradiol and 2.5 mg of dydrogesterone. This oral preparation should be taken continuously to treat climacteric symptoms in menopausal women. Femoston Low is in agreement with the recent recommendations for menopausal hormone replacement therapy, which give the preference to low dosage therapy whenever possible. The goals are to potentially minimize the risk of breast cancer, the danger of venous or arterial thrombosis and the glucose and lipid metabolic disturbances. Nevertheless, the preparation should efficaciously oppose to endometrial hyperplasia and yield a high degree of amenorrhea. PMID- 21638841 TI - [Therapeutic approaches to improve blood glucose control in a patient with type 2 diabetes on a metformin-sulfonylurea combination]. AB - Beyond lifestyle changes, the management of type 2 diabetes comprises the administration of oral glucose-lowering agents, especially the classical metformin-sulfonylurea combination. If such a dual oral therapy could not (any more) obtain an adequate glucose control, intensified management becomes mandatory. Several therapeutic approaches may be proposed at this stage, with some advantages and disadvantages of each of them. The present clinical case aims at illustrating such difficult therapeutic choice. We will provide the pro-contra arguments concerning each therapeutic alternative and describe the practical modalities of an appropriate management according to the patient's characteristics. PMID- 21638842 TI - [Institute of Clinical and Experimental Medicine--at the dawn of the fifth decade of successful existence]. PMID- 21638843 TI - [IKEM--scientific, political and other relationships of its foundation and to date development]. PMID- 21638844 TI - [Technique of implantation and experience with temporary mechanical cardiac support in right ventricular failure]. AB - AIM: Severe right heart failure remains unfrequent but fatal complication of cardiac surgical procedures. Implantation of temporary right ventricular assist device may be life-saving procedure in various situations of right heart failure as: heart transplantation, LVAD therapy and post-cardiotomy failure. The aim of the study is an introduction of the implantation technique and retrospective review of current experience with the method. MATERIAL AND METHODS: Since January 2007 isolated right ventricular assist device Levitronix CentriMag has been implanted in 16 patients. Patients were divided into 3 groups: post transplantation (post-Tx), post LVAD implantation (post-LVAD) and other cardiac procedures (OCP). Success rate of weaning from RVAD, 30-days mortality and major complications has been assessed. OUTCOMES: Distribution of implants in groups was: post-Tx 5 pts (31%), post-LVAD 6 pts (38%) and 5 in OCP group (31%). The mean support time was 12 days. Off-pump implantation was achieved in 9 pts. The device was successfully weaned in 13 (81%) patients. 30-days mortality occurred in 1 case only. CONCLUSION: Presented outcomes are encouraging for broader acceptance of the therapy. Excellent success rate has been reached in post-Tx and post-LVAD. This study emphasises decesive role of proactive approach in early indication of RVAD implantation for achieving satisfactory results. PMID- 21638845 TI - [Outcomes of combined mitral valve repair surgery and aortic valve replacement in IKEM Praha]. AB - AIM OF THE STUDY: We retrospectively analyzed long-term outcome of concomitant mitral valve repair and aortic valve replacement. METHODS: From 1996 to 2009 we performed mitral valve plasty with aortic valve replacement in 50 patients. Clinical and echocardiographic data were obtained from computer database and hospital records. Missing data were obtained through mailed questionnaire. We evaluated hospital mortality, long-term survival, thromboembolic and hemorrhagic complications and TR of 3+ on follow up echocardiography. RESULTS: Four patients who had previously undergone aortic valve surgery were excluded from the study. Aortic valve pathology was stenosis in 21 patients, regurgitation in 20 and 4 patients presented with mixed aortic disease. The etiology of mitral regurgitation was rheumatic in 6, non-rheumatic in 31 and infective in 6 patients. Aortic valve was replaced with mechanical prosthesis in 22 (mean age 59) and tissue prosthesis in 24 (mean age 71) patients. Additional surgical procedure was performed in 26 patients. Follow-up was 94% complete, with a mean duration of 51 months. Hospital mortality was 13%. Two and five year survival was 79% and 64% respectively. We noted one case of retroperitoneal hemorrhage and one stroke. We recorded 9 (19.6%) patients with residual TR of more than 3+ grade on follow up echocardiography. Out of 9 patients with residual TR, 3 were operated for rheumatic and 6 for non-rheumatic mitral valve disease. One patients underwent successful mitral valve replacement with mechanical prosthesis, 3 died and 5 are treated expectantly. CONCLUSION: We conclude that concomitant mitral valve repair with aortic valve replacement has high hospital mortality, excellent long-term survival and low complication rate. The durability of mitral valve repair in patients with rheumatic mitral valve disease is limited and replacement, rather that repair should be considered in this patient group. PMID- 21638846 TI - [Cardiosurgical procedures in patients refusing blood transfusions: IKEM outcomes]. AB - INTRODUCTION: Jehovah's Witnesses who require cardiac operation represent a specific challenge to the physicians. Members of this faith will not accept blood or blood products under any circumstances on the basis of religious grounds. Nevertheless cardiac operations belong to surgical interventions with potential severe bleeding and necessity of blood transfusions. THE AIM OF THE STUDY: The aim of this retrospective study was to analyze clinical data, operative and postoperative courses of patients operated at IKEM who refused blood transfusions. METHODS AND RESULTS: From January 1995 to August 2010, 73 Jehovah's Witnesses ranging in age from 19 to 82 years underwent cardiac surgery at our institute. Aortocoronary bypass were performed in 34 patients, valve surgery in 25 patients, 6 patients underwent concomitant aortocoronary bypass and valve surgery, 2 patients underwent aortocoronary bypass and resection of the left ventricle aneurysm and 2 patients underwent atrial septal defect repair and tricuspid valve anuloplasty. Ventricular septal sefect repair, atrial septal defect repair, Cor Cap device implantation and left ventricular epicardial electrodes implantation were performed in the other patients. Early 30-days mortality was 2.8 % (2 patients). CONCLUSION: We can conclude that cardiac surgery in Jehovah's Witnesses can be performed safety without blood transfusion and belongs to standard operating procedures at our institution. PMID- 21638847 TI - [Vascular complications in transcatheter aortic valve implantation in IKEM]. AB - AIM: Retrograde transfemoral arterial approach is the most common technique of transcatheter aortic valve implantation. Diameter of available catheters is the limiting factor for percutaneous usage. We currently use 18 French third generation Medtronic Core Valve system. We retrospectively analyzed procedure related complications in our patient cohort. MATERIAL AND METHODS: Transcatheter aortic vale implantation with 18 French Medtronic Core Valve was performed in 35 patients (23 females, 12 males) in between 12/2008 and 7/2010. RESULTS: Mean age was 81.4 +/- 6.1 years (range 69-92), mean logistics EuroSCORE was 19.3 +/- 8.9% (range 8-42), mean aortic valve gradient 59.8 +/- 19.8 mm Hg (range 30-86 mm Hg (mean indexed aortic valve area 0.37 +/- 0.11 cm. PMID- 21638848 TI - [Simultaneous transplantation of the kidney and the islets of Langerhans]. AB - Simultaneous kidney and islet transplantation is recent therapeutic alternative for diabetics with end-stage kidney disease, who are not acceptable for simultaneous pancreas-kidney transplantation. Islet transplantation has less complications but still worse long-term function compared to whole pancreas transplantation. PMID- 21638849 TI - [Liver transplantation in patients with portal vein thrombosis]. AB - AIM: The aim of the study was to perform a retrospective assessment in a group of patients with portal vein thromboses who underwent consecutive liver transplantation. MATERIAL AND METHODS: PVT was preoperatively diagnosed with ultrasound and CT portography. The follow up period was 1 to 6 years. Postoperative immunosuppressive medication was administered in combination with cyclosporin A, prednison and imuran upon initiation of the transplantation programme (8 patients). During the follow up period, the treatment protocol was replaced with FK 506, Cell Cept and prednisone. Liver biopsy was indicated when rejection was suspected. RESULTS: During 1996-2009, the team of authors performed a total of 740 liver transplantations in 303 female and 437 male subjects. The procedures included 703 primary procedures, 33 retransplantations and four second retransplantations. Out of the total, 57 recipients (7.7%) had portal vein obliteration. These subjects included 42 male and 15 females, their mean age was 52 (9-67) years. 62 liver transplantations were performed in these 57 patients. The following complications were recorded in the patient group: graft dysfunction in 10.5%, revisions for bleeding in 28%, hepatic artery thrombosis in 10.5%, portal vein rethrombosis in 1.8%, biliary complications in 17.5%, acute rejections in 19.3%. Perioperative mortality rate was 15.8%.The mean blood derivates requirement was 17.1 (0-425) erythrocyte transfusion units, 27.1 (0 132) frozen plasma transfusion units and 2.6 (0-20) thrombocyte transfusion units. CONCLUSION: The results of out retrospective study indicate that portal thrombosis is not a contraindication for liver transplantation. Current surgical techniques make liver transplantations in patients with complete splanchnic venous thrombosis possible. PMID- 21638850 TI - [Early biliary complications following liver transplantation]. AB - INTRODUCTION: Biliary complications (BC) after liver transplantation (LTx) are serious problems with an incidence rate of 6-35%. There are two types of BC, early and late. Early BC is observed for 30 days post LTx or during the entire period of the patient's first hospital stay after LTx. The main causes of early BC are ischemia of the biliary tree (the bile duct of the transplanted liver is supplied only from the right hepatic artery descendingly; the supply from the gastroduodenal artery is lacking) as well as implementation of the biliary anastomosis surgical technique. Treatment of BC is administered by a surgeon, a gastroenterologist and a radiologist. AIM: To evaluate early biliary complications after LTx PATIENTS AND METHODS: We reviewed patients with early BC after LTx from 4/2004 - 12/2009. We defined early BC as a complication that is present during the first 30 days post LTx or during the entire period of the patient's first hospital stay after LTx. RESULTS: We performed 384 LTx during the above-mentioned period. We carried out reconstruction of the biliary tree in the form of a choledochocholedochostomy (CDD) in 312 cases and in the form of a choledochojejunostomy (CDJ) in 72 cases. Sixty-eight patients had early BC (17.7%), 25 patients experienced biliary leak (6.5%), 40 patients had anastomotic stenosis (10.4%) and 3 patients (0.8%) experienced both complications (biliary leak and stenosis). Most complications were eliminated by ERCP with a papilosfincterotomy, a balloon dilatation of stenosis, and a biliary stent implant with repeated stent replacements (45 cases in total- all patients with biliary stenosis). Twenty-two patients were reoperated on (16x CDJ, 3x re-CDD, 2x suture of aberrant bile duct, 1x suture common bile duct) and 1 patient was treated by percutaneous transhepatic bilary drainage (PTD). The mortality rate was zero. CONCLUSION: BC after LTx continue to pose a serious surgical problem. The treatment of choice when dealing with BC is ERCP, which has more than a 70% success rate. If ERCP or PTD are not successful, or when biliary peritonitis is present, we perform a reanastomosis of the bile duct. The combined effort of the surgeon, gastroenterologist and radiologist is the most important factor for successful treatment. The incidence of BC after LTx at IKEM is similar to that of treatment centers all over the world. PMID- 21638851 TI - [First Czech and Slovak domino liver transplantation]. AB - INTRODUCTION: The study introduces the first international, Czech and Slovak Domino Transplantation performed based on a tight cooperation between the Czech and Slovak transplant centres. MATERIAL AND METHODS: The donor of the cadaveric graft was a young Slovak man with Familial amyloidotic polyneuropathy. The livers of these patients are the predominant source of circulating transthyretin, and liver transplantation is the only treatment available for the disease. The graft was obtained from heart beating deceased donor with brain death. The domino graft was then transplanted to Czech sixty-three years old man with hepatocellular carcinoma and liver cirrhosis based on HCV. RESULTS: Both recipients had an uneventful immediate postoperative course with early graft function. The Czech patient faced early HCV recurrence in a graft which necessitated the reduction of immunosuppressives. CONCLUSION: The domino liver transplantation represents a suitable way for addressing graft shortage and reducing waiting list time. A well organized cooperation between the two international centres is required to complete successful domino transplantation. PMID- 21638852 TI - [Development of pancreas transplantation surgical techniques in IKEM]. AB - A paper on the occasion of the fortieth anniversary of foundation of the Institute for Clinical and Experimental Medicine describes past, present and future trends in surgical technique of pancreas transplantation in the Czech Republic. PMID- 21638853 TI - [Immunosuppression following venous allografts transplantations--the authors' experience]. AB - AIM: Venous and arterial graft usage in vascular reconstructions was re discovered in connection with organ transplantation development. Allografts are employed in many clinics, however, uniform opinion on the use of immunosuppression after the procedure of venous graft transfer from a cadaveric donor, is still lacking. MATERIAL AND METHODS: The authors present their own group of patients who underwent vascular reconstructions, and in whom allogenic vein was used. The majority of indications for bypass procedures resulted from critical limb ischemia. Immunosuppressive medication was administered during the vascular procedure and, over the past several years, it purely consisted of tacrolimus monotherapy. RESULTS: In the group of 101 patients, no serious complications due to adverse effects of immunosuppression therapy were recorded. PMID- 21638854 TI - [Effect of biological glues on vascular wall in an experimental model of aortic dissection]. AB - INTRODUCTION: The use of biological glues and their application between the two dissection layers and into the anastomosis region is a common integral part of surgical management of thoracic aortic dissections. AIM: The aim of the experimental study was to assess and evaluate histopathological changes of vascular wall following deposition of the following three types of glue--GRF, Tissucol, Bioglue, based on qualitative and quantitative parameters. The secondary aim of the study was to assess dynamics of these changes depending on the glue effect duration and to formulate expected behaviour of the vascular wall during the time beyond the experimental period. METHODOLOGY: The dissection model was performed with pigs of the same gender and age, assigned to four groups. Different glues were used to close artificial infrarenal aortic dissections in Group 1-3, while direct suturing and no glue was used to close false lumini in Group 4. Samples of the dissected aorta were then collected at Month 1, 6 and 12 and then histologically examined. RESULTS: Upon assessment of the whole group of qualitative and quantitative parameters, the most significant changes in the smooth muscle histological picture were observed with the use GRF glue. The smooth muscle changes following the Bioglue application and, in particular, Tissucol glue application, are similar to changes observed in Group 4, where no glue was used. CONCLUSION: Based on the results, the authors present a hypothesis that, in a long-time horizont, vascular wall destructions, eventually redissections, are likely to occur more frequently in patients, in whom GRF glue is used. PMID- 21638855 TI - ["Red socks" in heat]. PMID- 21638856 TI - [From physical therapy to botulinum toxin: treatments in distressing spasticity]. PMID- 21638857 TI - [Dyspnea in the hypertensive patient. Why was diuretic therapy not effective?]. PMID- 21638858 TI - [Patient taking long-term steroids: monitoring bone density and blood glucose!]. PMID- 21638859 TI - [Dental care with local anesthesia: should the patient first discontinue ACE inhibitors?]. PMID- 21638860 TI - [Information about self-help groups: a must in every waiting room]. PMID- 21638861 TI - [Self-help groups for patients with psychic disorders or addiction and their relatives]. PMID- 21638862 TI - [Hypothyroidism]. PMID- 21638863 TI - [54-yr-old female with swollen joints, hemoptysis and hematuria]. PMID- 21638864 TI - [Antidepressants, antipsychotics and metabolic risks]. PMID- 21638865 TI - [Talking is gold and silence is silver]. PMID- 21638867 TI - [DBfK functionary: "I no longer recommend a career in nursing". DKI Spring Congress: nursing representatives deliver conflicting views on the Rosler reform]. PMID- 21638869 TI - [Diversity in ambulatory pediatric nursing. "It was like a trip around the world..."]. PMID- 21638870 TI - [Objective hermeneutic analysis of the nursing status. Understanding between the lines]. PMID- 21638871 TI - [Modular nursing rounds: innovative collection of key figures and reliable outcome quality measures. Detecting and solving problems]. PMID- 21638872 TI - [Person-centered case conference in nursing practice - key aspect of implementation. Recognizing and diffusing conflicts]. PMID- 21638873 TI - [Total conversion of the information and documentation system. Happy without paper]. PMID- 21638874 TI - [Professional communication structures by creating case conference. Developing solution according to plan]. PMID- 21638875 TI - [Fitness training after stroke. What impact does it have?]. PMID- 21638876 TI - [Effects on carers caring for residents who are limited in their everyday capabilities. A comparison of several measuring points in a pilot study]. AB - According to the section 87b SGB XI (German Social Security Statutes) care worker guidelines, care institutions can employ additional carers for residents who are permanently limited in their everyday capabilities. Studies on the stress experience while caring for the elderly who relate specifically to the additional care givers have, until now, not been available. Previous studies relate to trained nurses. It can be assumed that, if trained nurses already show typical stress reactions, the care givers could be all the more overburdened. For surveying the effects on care givers caring for residents who are limited in their everyday capabilities, an empirical data investigation using the Copenhagen Psychosocial Questionnaire (COPSOQ) was carried out on 30 care givers qualified according to the above mentioned guidelines in nine nursing homes administrated by Curanum AG. Additionally, the study took place in two residential homes of other administrators. As a comparison, 30 nurses were investigated. In the time span from the start of professional conduct until the end of the survey after six months, significant differences in the examined nurses were found only for the item Job Security (p < or = 0.05). In the group of section-87b-carers it was possible to show significant changes in the following items: possibilities for development (p 0.05), influence (p < or = 0.05), social support (p < or = 0.05), feedback (p < or = 0.05) and social-relationship p < or = 0.05). In summary, the psycho-social stress experiences of the section-87b-carers were determined to be considerably worse than those of the professionally qualified nurses, which could be due to the fact that section-87b-carers are less well trained than the professionally qualified nurses after their three years of training. The section 87b-carers continuously look after residents, who are considerably limited in their everyday capabilities, whereas the professionally qualified nurses look after a mixed group of residents. The study thus provides an indication that the use of section-87b-carers--at least from the viewpoint of the people concerned- leads to a tendency of overburdening and therefore, with the current training and accompaniment provided during employment, does not represent a reasonable and sustainable enhancement to everyday life in the nursing residents. PMID- 21638877 TI - [Information security in patient transfer]. PMID- 21638878 TI - [Preventing errors and proper network organization. Using opportunities of digital nursing documentation]. PMID- 21638879 TI - [Obtaining nursing care managers: experts develop an assessment center. Headhunting with a system]. PMID- 21638880 TI - [Early diagnosis of dysphagias after stroke. Nursing students learn to conduct a simple swallowing screening]. PMID- 21638881 TI - [Reflections on team nursing. Not all have to function identically]. PMID- 21638882 TI - ["Oath of confidentiality" in nursing. "I can not tell you, ask the doctor"!]. PMID- 21638883 TI - [From case to case. Advantage through technology?]. PMID- 21638884 TI - [Dysuria as the leading symptom. When urination malfunctions]. PMID- 21638885 TI - [Medicalization and dependence on the health system]. PMID- 21638886 TI - [What do nurses think of pain associated with vascular functions?]. AB - One of the most common techniques used by nurses in carrying out their clinical practice is the vascular puncture, commonly associated with pain bearable for the patient to a greater or lesser extent. For the past few years have been appearing in the pharmaceutical industry a series of local anesthetics can be used for vascular puncture to minimize or alleviate the pain. The increasing interest by nurses in this field is evidenced by the growing number of scientific articles published in this discipline, included in the Intravenous Therapy. During the Fourth National Congress of the Association of Intravenous Therapy Teams held in 2010 in Valencia went over to the Congress a survey to assess their knowledge and concerns about pain control in the punctures and the use of local anesthetics in the same. PMID- 21638887 TI - [Modification of body temperature as clinical therapeutics. Hypothermia]. AB - The application of cold therapy is called thermotherapy Can distinguish two major forms: local and general. At the local level is widely used in the field of physical rehabilitation, rheumatology and various surgical specialties. However the evidence on issues relating to its potential benefits, physiological reason underlying its action, or application form is not sufficiently supported. Regarding the application of cold techniques, higher expectations are focused on hypothermia induced or therapeutic, for its significant neuroprotective effects after ischemia secondary to cardiac arrest. Interest in hypothermia begins in the third decade of the twentieth century the first scientific report on induced hypothermia was published in 1945 and described its application in patients with TBI. In the following decades appeared critical research on their application, but also great shadows of its benefits. The development of protocols and the great advances in the care provided in ICUs, have increased the capacity to prevent and control the side effects of the application of this therapy Based on the literature review, describing the main uses of therapeutic hypothermia, the major indications of these techniques and the evidence of its benefits and complications arising from their use. PMID- 21638888 TI - [Strategies for patient safety in medication management and administration]. AB - Clinical safety is one of the greatest challenges currently facing healthcare organizations. Is a key element in the quality of care processes, care processes among which noted, by its impact, the medication management and administration. All health professionals need, in developing their professional competence, strategies to improve safety in medication management and to incorporate the prevention of errors and adverse events in their training programs and assistance: only from proper planning will be approached for excellence in clinical care and safety. PMID- 21638889 TI - [Anesthetic gases and nurses in operating room]. AB - Summary The growing number of women of childbearing age joining the job market and remaining work active during pregnancy determines the importance of preventing the potential hazards of the workplace during pregnancy and while breastfeeding. We must take into account that regular working practices may not be acceptable for women during pregnancy or while breastfeeding. One of the daily risks of operation theatre nurses is exposure to inhaled anaesthetics used during surgery In most cases, nurses are unaware if exposure is permitted during pregnancy or while breastfeeding. In this article, it is intended to publicize these risks, as well as the ways to prevent, taking into account the existing health and safety laws. PMID- 21638890 TI - [Design and implementation of management strategies for the improvement in the implementation of the warnings of isolation in a pediatric hospital]. AB - INTRODUCTION: children's Hospital Vail d'Hebron in 2007 children admitted 7086, requiring isolation 440 (6.2%). The purpose of this study was to raise the overall level of adherence to the basic precautions of isolation. The nursing management by objectives allowed us to raise as a management strategy in pediatric inpatient units, improved compliance with isolation measures, multidisciplinary goal framed the need for pediatric patient safety OBJECTIVES: to achieve compliance with the protocol of isolation described in 80-90% of patients. METHODOLOGY: it provides all pediatric units a dossier of consultation on the types of insulation and measures to follow, informative posters in the areas of work and training sessions. It develops and implements a standardized checklist for the proper performance of isolation measures in two of the six child health. RESULT: working in an MBO has focus and unify the resources for the improvement in the performance of isolates. In the units where they have implemented the checklist has found a fulfillment of isolation measures in 96% of cases. CONCLUSIONS: this experience has allowed us to confirm that the strategies carried out by monitoring compliance contribute to a more effective isolation measures hospital and export the model of implementation of the checklist to other pediatric units. PMID- 21638891 TI - [Appearance of hematomas after blood extractions related with needle gauge related]. AB - We want to know the method that less damage produced in the patient when making a venous blood extraction. The objective of this study is to compare the system of extraction that provokes less hematomas and reduces pain in the patients. We studied 175 patients; the variables studied were age, duration of the technique, occurrence of haematoma, pain perceived according to numerical scale of the pain. The study shows that does not exist relation between the appearance of haematomas and the catheter with which the venous puncture is realized, at least, not between catheters of 21 and 20 gauge. It was noted that the patients feel less pain when needles of calibre 21 gauge (butterflies) were used. The extraction time is lightly bigger with this system, but if we want to offer some quality care, we must perform the extraction of blood with the butterfly device (needles of 21 G). PMID- 21638892 TI - [Tetanus in Spain. Mortality study along the XX century to the present]. AB - The present study aims to highlight the severity of a disease now almost eradicated through vaccination, tetanus, but that was a high mortality during much of the last century. PMID- 21638893 TI - [Human beings are always changing, new, and different beings. Interview by Maria Jesus Nadal]. PMID- 21638894 TI - [Neoplastic wounds]. PMID- 21638895 TI - [Let's overcome the false belief that it is not the right moment, that it is better to protect ourselves and wait for the calm weather. Interview by Maria Jesus Nadal]. PMID- 21638896 TI - [A will to child neurologists]. PMID- 21638897 TI - [A 1-year-old girl with regression, dysphagia and dyspnea]. PMID- 21638898 TI - [Future perspectives of medical journals in pediatrics]. PMID- 21638899 TI - [Update on multimodal neurosurgical management of dystonias]. AB - The neurosurgical treatment of dystonia has progressed markedly since the introduction of deep brain stimulation of the globus pallidum interna. However, dystonia is not a single disorder but comprises various types and causes, and it is true that deep brain stimulation cannot cover the complex nature of dystonia. Depending on the distribution of symptoms and causes, we have to consider other surgical managements such as thalamotomy, peripheral denervation, and intrathecal baclofen. Such a multi-modal strategy has enabled us to treat and even cure many patients with dystonias. No treatment other than various neurosurgical approaches yields better results in the management of dystonias. In this sense, we are now at a stage where we should regard dystonia as a neurosurgical disorder in terms of treatment. PMID- 21638900 TI - [The perspective of perinatal brain injury]. PMID- 21638901 TI - [Aquaporin from basic to clinical medicine: roles in brain edema]. PMID- 21638902 TI - [Illusion or reality? Acute cerebral injury and advanced magnetic resonance]. PMID- 21638903 TI - [Stem cell therapies CNS diseases--can they save babies with brain injuries?]. PMID- 21638904 TI - [Prenatal brain damage from obstetrical point of view]. PMID- 21638905 TI - [The present situation and creating new system for children with severe motor and intellectual disabilities, medical care dependent group and their parents]. PMID- 21638906 TI - [Psychologist in child neurology]. PMID- 21638907 TI - [Recent topics of children's brain death in Japan]. PMID- 21638908 TI - [Short-term and long-term efficacy of topiramate in refractory generalized epilepsy of children]. AB - Topiramate (TPM) was administered to 25 children with intractable generalized epilepsy. A > or =50% decrease in seizure frequency was observed in 56% and 45% of children at two months and one year after initiation of TPM therapy, respectively. However, efficacy of TPM for Lennox-Gastaut syndrome was low. TPM therapy was discontinued in five of 25 children at 3-5.5 months due to lack of efficacy or aggravation of seizures. No serious adverse effects were observed during TPM therapy. The present study revealed that TPM has clinical efficacy in the treatment of children with intractable generalized epilepsy. PMID- 21638909 TI - [Fatal esophageal hemorrhage in patients with severe motor and intellectual disabilities complicated with severe scoliosis, during catheter placement in esophagus]. AB - In patients with severe motor and intellectual disabilities (SMID), fatal esophageal hemorrhage including penetration of the descending aorta, has often been reported. We experienced 2 patients with severe scoliosis who developed esophageal hemorrhage during catheter placement in the esophagus. We compared chest CT images of these cases with those of 38 SMID patients in our hospital. As a result, a few patients showed esophagus bending to the left of the descending aorta while the vertebral body curved to the right and the mediastinum shifted to the left. It is suggested that there is a risk of esophageal hemorrhage caused by catheter stimulation, when a catheter is placed in the esophagus in such patients. It is therefore necessary to use the thinnest and softest catheter possible and to consider the indications for gastrostomy. PMID- 21638910 TI - [Three cases of Fanconi syndrome associated with valproate sodium treatment]. AB - Valproate sodium (VPA) is a commonly used antiepileptic drug. However, various side effects, including liver dysfunction, thrombocytopenia, anorexia, hyperammonemia, and pancreatitis have been reported in association with the administration of VPA. Recently, renal Fanconi syndrome associated with VPA treatment has occasionally been reported. However, the mechanisms and detailed characteristics of this adverse effect remain unknown. We herein report three cases of Fanconi syndrome associated with VPA treatment. All of these patients were severely disabled children, who had been previously treated with multiple antiepileptic drugs, and also required tube feeding. The possible risk factors of Fanconi syndrome in these three cases were similar to those previously reported in the literature. In addition, all three patients developed Fanconi syndrome after the onset of bacterial infections. Before developing Fanconi syndrome, hypouricemia was observed in all three and an increased urinary level of beta2 microglobulin (beta2MG) was also noted in one of the patients. None of these patients had hypophosphatemia. Two patients had an appropriate serum VPA level, while the other had an inappropriately high level. We therefore recommend that severely disabled children receiving multiple antiepileptic drugs and tube feeding be periodically checked for urinary beta2MG and uric acid, especially during the course of any infectious episodes. PMID- 21638911 TI - [The preliminary study on early signs of high-functioning developmental disorder]. PMID- 21638912 TI - [Physicians are needed in IT development]. PMID- 21638913 TI - [The fall of a star]. PMID- 21638914 TI - [Aging with "compressed morbidity" one more illusion. The morbidity during aging is more likely to have expanded]. PMID- 21638915 TI - [Anxiety is our heritage and we should administer it well]. PMID- 21638916 TI - [Psychiatry and primary health care have common responsibility]. PMID- 21638917 TI - [Common early-onset diseases]. PMID- 21638918 TI - [Generalized anxiety can be treated effectively]. PMID- 21638919 TI - [Panic syndrome--evil circle of incorrectly interpreted bodily signals]. PMID- 21638920 TI - [Again and again--the obsessive circle can be broken]. PMID- 21638921 TI - [Social phobia--no common shyness]. PMID- 21638922 TI - [New research tracks pave the way for better therapy]. PMID- 21638923 TI - [Itching nodules after Havrix vaccination]. PMID- 21638924 TI - [Fatigue and pseudodiagnostics]. PMID- 21638925 TI - [Vitamin D deficiency--who's responsibility?]. PMID- 21638926 TI - [Losing a parent is fatal: health care must assure greater responsibility also for the relatives]. PMID- 21638927 TI - Insect motion detectors matched to visual ecology. AB - To detect motion, primates, birds and insects all use local detectors to correlate signals sampled at one location in the image with those sampled after a delay at adjacent locations. These detectors can adapt to high image velocities by shortening the delay. To investigate whether they use long delays for detecting low velocities, we compared motion-sensitive neurons in ten species of fast-flying insects, some of which encounter low velocities while hovering. Neurons of bee-flies and hawkmoths, which hover, are tuned to lower temporal frequencies than those of butterflies and bumblebees, which do not. Tuning to low frequencies indicates longer delays and extends sensitivity to lower velocities. Hoverflies retain fast temporal tuning but use their high spatial acuity for sensing low-velocity motion. Thus an unexpectedly wide range of spatio-temporal tuning matches motion detection to visual ecology. PMID- 21638928 TI - [Expression of platelet-derived growth factor receptor alpha and its ligand PDGF- A in breast cancer]. AB - AIM: To investigate the potential significance of platelet-derived growth factor receptor-alpha (PDGFR-alpha) and platelet-derived growth factor A (PDGF-A) expression in mammary carcinomas, and analyze its correlation with the lymph node metastasis and the expression of PDGF-A. METHODS: Used immunohistochemistry to detect the protein expression of PDGFR-alpha and PDGF-A in paraffinembedded breast carcinomas. RESULTS: The expression of PDGFR-alpha and PDGF-A were observed in 51.7% and 61.7% in the breast carcinomas, respectively and showed an association with lymph node metastasis (P < 0. 05). A correlation was also found with the expression of PDGFR-alpha and PDGF-A (P<0.05). CONCLUSION: PDGFR-alpha is expressed in invasive breast carcinomas and is associated with biological aggressiveness. PMID- 21638929 TI - Chest X-ray quiz. PMID- 21638930 TI - [Clinic research of the effect of bFGF on human degenerated nucleus pulposus cells]. AB - AIM: To determine the effect of bFGF on extracellular matrix and gene expression on the human de-generated nucleus pulposus cells. METHODS: The degenerated intervertebral disc cells were divided into 5 groups. A group: adding 100 pg/L bFGF, B group: adding 200 pg/LbFGF, C group: 500 pg/L bFGF, D group: 1000 pg/L bF GF, E groups: control group, without interfering factors. The type II collagen and GAG mRNA expression were test, type II collagen content and glycosaminoglycan content of the supernatant were test. RESULTS: bFGF stimulated de-generated nucleus pulposus cells, collagen II, GAG mRNA, collagen type II and glycosaminoglycan expression of the extracellular matrix were increased significantly on the 7 days compared with the control group (P < 0. 05). type II collagen and GAG mRNA were significantly lower than the control group on 14 days, 21 days (P < 0.05). However,collagen type II and glycosaminoglycan expression of the extracellular matrix on 14 days, 21 days were still higher than normal (P <0.05). CONCLUSION: bFGF increase GAG and type II collagen mRNA expression, increased extracellular matrix type II collagen and GAG expression in short-term. PMID- 21638931 TI - [The effect of protein metabolism and immunologic function of glutamine after operation in elder gastrointestinal tumor]. AB - AIM: To explore the effect of protein metabolism and immunologic function of glutamine after operation in elder gastrointestinal tumor. METHODS: Form march 2007 to 2010, 87 cases of elder gastrointestinal tumor were given parenteral nutrition and glutamine 0.6 g/( Kg x d).The period of treatment were 8 days. IgA, IgG, IgM were CD4, measured by single immunodiffusion, CD3(+), CD4(+), CD8(+), CD4(+) /CD8(+) were measured by immunohistochemical method, and the index(Alb, PAB, TF, nitrogen equilibrium) were monitored the proteid catabolism distribution. RESULTS: After the treatment, CD3(+), CD4(+), CD8(+), CD4(+)/CD8(+), IgG, IgA, IgM were evidently declined( P <0.05). Alb, PAB, TF were evidently declined in 4 days postoperatively (P < 0.05), the restore were more obvious in 8 days postoperatively (P < 0. 05). Nitrogen equilibrium was worse in the early postoperative and the restore were more obvious in 8 days postoperatively (P < 0.05). CONCLUSION: Glutamine can improve patient's nutrition, enhance their immunologic function. PMID- 21638932 TI - Comparative health politics: the United States and the United Kingdom. PMID- 21638933 TI - [Multiprobe fluorescence in situ hybridization panel in detection of the common cytogenetic abnormalities of acute myeloid leukemia]. AB - AIM: To evaluate the value of multiprobe Fluorescence in situ hybridization (FISH) panel in detection of the common cytogenetic abnormalities in acute myeloidleukemia( AML). And to investigate its association with clinical diagnosis, chemotherapy and prognosis. METHODS: Using the multiprobe AML/MDS panel designed to detect upto eight different FISH probes, which was for AML1/ETO transfusion gene, PML-RARalpha transfusion gene, CBFbeta/MYH11 transfusion gene, MLL breakapart, P53 deletion,Del(5q), Del(7q), Del(20q), 40 cases of AML were investigated. The conventional karyotype analysis and the in-formation about the treatment responses were also used for assessing. RESULTS: 22 of the 40 AML cases were found to carry 7 types of cytogenetic abnormalities by multiprobe FISH panel including AML1/ETO transfusion gene, PML-RARa transfusion gene, MLL breakapart, P53 deletion, Del (5q), Del7q and trisomy 8. However conventional karyotype analysis only discovered 11 cases with the corresponding cytogenetic abnormalities, the positive ratio was 57.5% in multiprobe FISH panel higher than that in karyotype analysis (27.50%). Patiens with AML1/ETO or PML-RARa transfusion gene are easily to reach CR in the first induction chemotherapy, while the Del(7q), MLL breakapart, complex cytogenetic abnormalities may indicate poor prognosis. CONCLUSION: Mutiprobe FISH panel is more rapid, accurate and effective for detecting the common cytogenetic abnormalities in AML, compared with the conventional karyotype analysis and common FISH analysis. PMID- 21638934 TI - [The study on the changes of serum IL- 6, TNF-alpha and peripheral blood T lymphocyte subsets in the pregnant women during perinatal period]. AB - AIM: To study the change law of serum IL-6, TNF-alpha and peripheral blood T lymphocyte subsets in the pregnant women during perinatal period. METHODS: 100 pregnant women in our hospital from November 2009 to October 2010 were selected as research object, and the serum IL-6, TNF-alpha and peripheral blood T lymphocyte subsets be-fore and at labor onset occurring, after delivery at the first and third day were analyzed and compared. RESULTS: According the study, the serum IL-6 and TNF-aat labor onset occurring were higher than those before labor onset and af-ter delivery at the first and third day , the CD3(+), CD4 (+), CD8(+) and CD4/CD8 decreased first and then increased, all P < 0. 05, there were significant differences. CONCLUSION: The changes of serum IL-6, TNF-alpha and peripheral blood T lymphocyte subsets in the pregnant women during perinatal period has a regular pattern, and it is worthy of. PMID- 21638935 TI - Delivery system reform tracking: a framework for understanding change. AB - The health care delivery system is changing rapidly, with providers forming patient-centered medical homes and exploring the creation of accountable care organizations. Enactment of the Affordable Care Act will likely accelerate these changes. Significant delivery system reforms will simultaneously affect the structures, capabilities, incentives, and outcomes of the delivery system. With so many changes taking place at once, there is a need for a new tool to track progress at the community level. Many of the necessary data elements for a delivery system reform tracking tool are already being collected in various places and by different stakeholders. The authors propose that all elements be brought together in a unified whole to create a detailed picture of delivery system change. This brief provides a rationale for creating such a tool and presents a framework for doing so. PMID- 21638936 TI - [Human chorionic gonadotrophin on the expression of epidermal growth factor in rat renal tissues]. AB - AIM: To investigate hCG on the expression of epidermal growth factor (EGF) in renal tissues. METHODS: 20 male SD rats weighed 300 - 350 g were injected hCG 200 IU/d for 7 days, and 20 male rats were injected sa-line at equal volume as a control. Kidney tissues were per-formed with immunohistochemical staining to evaluate the distribution of EGF and quantitative PCR (Q-PCR) was used to evaluate the EGF mRNA. RESULTS: Immunohistochemtry showed that the positive rate of EGF in experimental group was significantly higher than that in control group(P<0.01), but Q-PCR results showed no significant difference between two groups ( P > 0. 05). CONCLUSION: hCG increased the concentration of EGF by maturing pre-pro-EGF not expressing EGF from de novo synthesis. PMID- 21638937 TI - Gene expression profiling in women with lymph-node-positive breast cancer to select adjuvant chemotherapy treatment. PMID- 21638938 TI - Wearable cardioverter-defibrillator as a bridge to implantable cardioverter defibrillator treatment. PMID- 21638939 TI - Special report: positron emission tomography for the indication of post-treatment surveillance of cancer. PMID- 21638940 TI - Low-level laser therapy for carpal tunnel syndrome and chronic neck pain. PMID- 21638941 TI - Radiofrequency ablation of nondysplastic or low-grade dysplastic Barrett's esophagus. PMID- 21638942 TI - Epidermal growth factor receptor mutations and tyrosine kinase inhibitor therapy in advanced non-small-cell lung cancer. PMID- 21638943 TI - Proton beam therapy for non-small-cell lung cancer. PMID- 21638944 TI - Percutaneous tibial nerve stimulation for the treatment of voiding dysfunction. PMID- 21638945 TI - Proceed with caution. Lean year expected for hospital construction. PMID- 21638946 TI - Clean design: keeping patient protection in mind. PMID- 21638947 TI - Successful measures: benchmarking clinical engineering performance. PMID- 21638949 TI - Ascending hope. PMID- 21638948 TI - Compliant compounding. Meeting USP 797 pharmacy regulations. PMID- 21638950 TI - Green + Greener. Hospitals embrace environmentally sustainable practices, though laggards remain. PMID- 21638951 TI - Lab work. A 10-step process for laying out an efficient clinical laboratory. PMID- 21638952 TI - Cost conscious. Finding design and construction efficiencies throughout the process. PMID- 21638953 TI - Cleaning up. Trends in housekeeping equipment and supplies. PMID- 21638954 TI - Gas flow. Developing a medical gas O&M program. PMID- 21638955 TI - Nature's calming influence. PMID- 21638956 TI - Comparative study of physico-chemical properties of MTA-based and Portland cements. AB - The purpose of this investigation was to evaluate the physicochemical properties of gray and white structural and nonstructural Portland cement, gray and white ProRoot MTA and MTA BIO. The water/powder ratio, setting time, solubility and pH (hydrogen-ion potential) changes of the materials were evaluated. Tests followed specification #57 from the American National Standard Institute/American Dental Association (2000) for endodontic sealing materials and pH was determined by a digital pH meter. The test results were statistically analyzed by variance analyses for global comparison and by the complementary Tukey's test for pairwise comparisons (5%). Considering the water/powder ratio, no significant difference (p > 0.05) was observed among the cements. MTA BIO (33.10 +/- 2.30) had the lowest setting time (p < 0.05), gray ProRoot MTA (10.10 +/- 2.70) had the highest (p < 0.05). White nonstructural Portland cement (2.55 +/- 0.08) had the highest solubility (p < 0.05), while gray ProRoot MTA (1.03 +/- 0.12) had the lowest (p < 0.05), although all materials showed solubility values in compliance to ANSI/ADA. No difference (p > 0.05) was observed among materials when considering pH evaluation. The pH levels were highly alkaline immediately after immersion in solution, remaining stable throughout the test period. The authors conclude that the cements had similar water/powder proportions. MTA BIO had the shortest setting time and gray ProRoot MTA had the lowest solubility. All cements had similar behavior in the pH analysis. PMID- 21638957 TI - Relationship between psychological factors and symptoms of TMD in university undergraduate students. AB - Temporomandibular disorders is a collective term used to describe a number of related disorders involving the temporomandibular joints, masticatory muscles and occlusion with common symptoms such as pain, restricted movement, muscle tenderness and intermittent joint sounds. The multifactorial TMD etiology is related to emotional tension, occlusal interferences, tooth loss, postural deviation, masticatory muscular dysfunction, internal and external changes in TMJ structure and the various associations of these factors. The aim of this study was to evaluate the prevalence of the relationship between signs of psychological distress and temporomandibular disorder in university students. A total 150 volunteers participated in this study. They attended different courses in the field of human science at one public university and four private universities. TMD was assessed by the Research Diagnostic Criteria (RDC) questionnaire. Anxiety was measured by means of a self-evaluative questionnaire, Spielberger's Trait State anxiety inventory, to evaluate students'state and trait anxiety. The results of the two questionnaires were compared to determine the relationship between anxiety levels and severity degrees of chronic TMD pain by means of the chi-square test. The significance level was set at 5%. The statistical analysis showed that the TMD degree has a positive association with state-anxiety (p = 0.008; p < 0.05) and negative with trait-anxiety (p = 0.619; p < 0.05). Moreover a high TMD rate was observed among the students (40%). This study concluded that there is a positive association between TMD and anxiety. PMID- 21638958 TI - Anticariogenic activity of the active fraction from Isertia laevis against S. mutans and S. sobrinus: comparison of two extraction methods. AB - Dental caries is considered a multi-factorial, infectious, chronic, localized, post-eruptive, transmissible disease that leads to the destruction of dental hard tissue. The recognition of Streptococcus mutans as the major bacterial species involved in dental caries has led to the implementation of prevention and control measures for eliminating or reducing it in oral cavity. The main goal of research on medicinal plants is the search for substances or compounds with antimicrobial activity. The aim of this study was to evaluate the antimicrobial activity of fractions obtained by two methods from Isertia laevis against S. mutans and S. sobrinus. The plant material was collected in Medina (Colombia), at an elevation of 550 meters above sea level. From the ethanol extract of leaves of I. laevis, fractions were obtained by two methods: extraction by column vacuum chromatography (CVC) and extraction by continuous liquid/liquid partitioning (CLLP). The evaluation of the antimicrobial activity of fractions against S. mutans and S. sobrinus was performed by well diffusion and bioautography assays. From the CVC technique, only the methanol and methanol-dichloromethane fractions showed activity against S. mutans and S. sobrinus, with a minimum inhibitory concentration of 2 mg/well. From the CLLP technique, only the dichloromethane fraction showed activity against both microorganisms, with a minimum inhibitory concentration of 1 mg/well. Compounds C1 and C2 were isolated from the three active fractions, and showed a minimum inhibitory concentration of 0.4 mg/well for S. mutans and S. sobrinus, with zones of inhibition measuring 6.5 and 6.2 mm, respectively. IN CONCLUSION: 1) the three active fractions of I. laevis showed activity against S. mutans and S. sobrinus, 2) compounds C1 and C2 were presen equally in the three active fractions showing activity against the two bacteria, 3) compounds C1 and C2 may be triterpenoid and/or steroidal saponin structures, and 4) the two extraction methods lead equally to obtaining the active fractions. PMID- 21638959 TI - Amoxicillin/metronidazole or scaling and root planing in the treatment of chronic periodontitis. AB - The aim of the present study was to evaluate the treatment with amoxicillin or metronidazole in comparison to scaling and root planing in the treatment of chronic periodontitis. Randomised clinical trials were searched in the databases MEDLINE, EMBASE, SciELO, Cochrane and Scopus from 1989 to 2010. The search started with 2895 articles. From this initial number of articles, 10 publications were selected and included in the study according to fixed criteria. Studies included adult patients of both sexes aged between 21 and 80, diagnosed with chronic periodontitis and treated with amoxicillin and/or metronidazole or scaling and root planning. From each article, details were abstracted relating to sample size, design, sex, age, oral hygiene habits, the exposure to drug (doses, schedule), and results such as clinical effect, analysis methods, stratification variables. CONCLUSION: this meta-analysis showed absence of statistically significant difference between the effects studied. PMID- 21638960 TI - Relationship between overweight-obesity and periodontal disease in Mexico. AB - The aim of this study was to examine the association between overweight-obesity and periodontal disease in subjects who attended the clinic of Periodontics, Faculty of Dentistry, San Luis de Potosi, Mexico. This was cross-sectional study involving 88 subjects--60 without overweight-obesity and 28 with overweight obesity. The following clinical parameters were evaluated: dental bacterial plaque, index of calculus, gingivitis, probing depth and periodontal disease index (PDI). When comparing the group of subjects with overweight-obesity to the control, there were statistically significant differences in the variables calculus (p = 0.0015), gingivitis (p = 0.0050) and periodontal disease (p = 0.0154). Regarding the logistic regression analysis, the dependent variable was subjects with and without overweight-obesity and the independent variables were sex, age and periodontal disease. We found statistically significant differences (p = 0.0162) with OR = 3.16 in periodontal disease. Periodontal disease showed statistically significant differences in the group of subjects with overweight obesity. The oral health of subjects with overweight-obesity should be supervised and checked in order to prevent oral alterations. PMID- 21638961 TI - Hydrophobicity test in mutans streptococci. AB - Kinetic hydrophobic measurements were performed by confronting 40 mutans streptococci from thirty 10- to 20-year-old patients with 200 ml hexadecane (Sigma). Fourteen patients had high dental caries risk (Group A), dmft + DMFT > 5 with 3 or more active caries, and 16 had low dental caries risk (Group B), dmft + DMFT < 3 without active caries. Twenty bacteria from Group A and 20 bacteria from Group B were typed using De La Higuera's procedure and confirmed by API strip (bio-Merieux). From the 14 patients in Group A we obtained 12 S. mutans (8 hydrophobic/4 non-hydrophobic), 5 S. sobrinus (4 hydrophobic/1 non-hydrophobic) and 3 S. rattus (hydrophobic). From the 16 patients in Group B we obtained 11 Streptococcus mutans (10 non-hydrophobic/1 hydrophobic), 7 Streptococcus sobrinus (6 non-hydrophobic/ 1 hydrophobic) and 2 Streptococcus rattus (hydrophobic). Patients with high dental caries risk have a higher prevalence of hydrophobic bacteria than patients with low dental caries risk (p = 0.0003). All typed S. rattus were hydrophobic. PMID- 21638962 TI - Effect of endodontic sealer and resin luting strategies on pull-out bond strength of glass fiber posts to dentin. AB - The aim of this study was to evaluate the influence of eugenol-containing endodontic sealers and luting strategy on the pull-out bond strength of glass fiber posts to dentin. Sixty-four bovine incisors were randomly assigned into two groups of 32 specimens each for obturation procedure with gutta-percha only, or with Pulp Canal Sealer EWT Subsequently, the roots were prepared for the fiber post Reforpost and all specimens of each endodontic sealing procedure were allocated to four groups (n = 8), according to the strategies for post cementation: A) Single Bond 2 and RelyX ARC; B) All Bond 2 and C&B cement; C) All Bond 2 and RelyX ARC; D) Single Bond 2 and C&B Cement. The posts were cemented immediately after the endodontic treatment. The pull-out test was performed at a cross-head speed of 0.5 mm/min in a universal testing machine (EMIC). Data (Kgf) were submitted to a two-way ANOVA and Tukey test (p < or = 0.05). The eugenol based sealer did not influence the pull-out bond strength of fiber posts regardless of the luting strategy. RelyX ARC showed higher bond strength than C&B Cement when used with Single Bond 2 adhesive system, when the eugenol-based sealer was present. All Bond 2, when associated to all cements studied, promoted similar bond strength, regardless of the eugenol-containing endodontic sealer In conclusion, eugenol-containing sealer did not influence the pull-out bond strength values of the resin luting systems for glass fiber post cementation. The bond system and resin cement association from the same manufacturer had similar bond strength values for dentin. PMID- 21638963 TI - Comparative inter-institutional study of stress among dentists. AB - Dentistry is considered to be a stressful profession due to different factors caused by work, representing a threat to dentists'health. The objectives of this work were to identify and compare chronic stress in dentists among the different health institutions and the association of stress with risk factors. The study in question is observational, transversal and comparative; 256 dentists were included, distributed among five public health institutions in the city of Guadalajara, Jalisco, Mexico, namely: the Mexican Institute of Social Security (IMSS), the Ministry of Health (SS), the Integral Development of the Family (DIF), the Social Security Services Institute for the Workers (ISSSTE) and the University of Guadalajara (U. de G) Data were obtained by means of the census technique. Stress was identified using the Stress Symptoms Inventory and the statistical analysis was performed using the Odds Ratio (O.R.) and the chi-square statistic. From the total population studied, 219 subjects presented high levels of chronic stress and 37, low levels. In the results of comparative analysis, significant differences were found between IMSS and U. de G and likewise between IMSS and SS. However, in the analysis of association, only U. de G was found to be associated with the high level of chronic stress. PMID- 21638964 TI - Prevalence of signs and symptoms of temporomandibular disorders in children in the State of Puebla, Mexico, evaluated with the research diagnostic criteria for temporomandibular disorders (RDC/TMD). AB - The aim of this work was to describe the prevalence of signs and symptoms of temporomandibular disorders (TMD) in children of the State of Puebla, Mexico. A descriptive observational study was performed. After calculating sample size, children who met the following selection criteria were included: registered at an official elementary school, either sex, ages between 8 and 12 years, who accept to participate in the study and whose parents have signed the informed consent forms. The Research Diagnostic Criteria for Temporomandibular Disorders were used by calibrated researchers (Kappa 90) under the same conditions. Descriptive statistics were applied by using SPSSv15 software. The study included 235 children, 129 (54.9%) female and 106 (45.1%) male, of average age 9.31 + 1.2 years. Prevalence of signs and symptoms was 33.2%, and predominately muscular (82%), 48.1% showed signs of muscular pain and 19.1% joint pain. 63.4% showed signs of alteration in the mouth opening pattern, 39.1% presented joint sounds on opening or closing the mouth and 20.4% on mandibular excursions. The high prevalence of signs and symptoms related to temporomandibular disorders, in particular in children with mixed dentition, shows the importance of TMD evaluation during this period, when morphological changes associated to growth and craniofacial development prevail. PMID- 21638965 TI - A preliminary study of enamel remineralization by dentifrices based on Recalden (CPP-ACP) and Novamin (calcium-sodium-phosphosilicate). AB - The purpose of this study was to investigate the enamel remineralization potential of two toothpastes, one of which was based on Recaldent (CPP- ACP) and the other on NovaMin (Calcium-sodium-phosphosilicate). Human permanent molar teeth were subjected to three consecutive demineralization cycles. These cycles were followed by remineralization of the experimental groups by toothpastes containing Recalden and NovaMin respectively. The samples were analyzed by Scanning Electron Microscope, (SEM) and energy-dispersive X-ray spectroscopy analysis (EDX). Extensive demineralization was noted in the control group (without remineralization) while the groups treated with the dentifices demonstrated various degrees of remineralization, as shown by formation of different types of deposits on the enamel surface. The EDX analysis showed increased amounts of Ca, P Si and Zn in the enamel of the experimental groups, compared to the control one. Toothpastes containing Recalden and especially NovaMin have the potential to remineralize enamel, a property which might be important in finding a substitute to pit and fissure sealing. PMID- 21638966 TI - Side effects of cyclosporine-A treatment in rats: gingival overgrowth and early hyperglycemia. AB - Gingival overgrowth is an adverse side effect of cyclosporine A (CsA) in the treatment of transplanted patients. The purpose of this study was to evaluate the effects of CsA on new-onset diabetes mellitus and gingival overgrowth in rats, by measuring collagen, nitric oxide and microvascular permeability. Blood glucose level, collagen, nitric oxide level and vascular permeability were determined. Blood glucose level increased significantly from 6.5 +/- 0.9 for the control group to 15 +/- 1.2, 17 +/- 1.2 and 21.6 +/- 1.6 mM/L at 1, 4 or 8 weeks of CsA treatment, respectively. Collagen (ug HO Proline/mg p) increased significantly from 2.5 +/- 0.5 for the control group to 4.2 +/- 0.8, 5.9 +/- 0.6 and 7.3 +/- 0.8 at 1, 4 or 8 weeks of CsA treatment, respectively. Vascular permeability was 10.3 +/- 1.2 for the control group and 15 +/- 1; 17.2 +/- 1.3, and 22.1 +/- 2.1 ug EB/g T; at 1, 4 or 8 weeks of CsA treatment, respectively Nitric oxide level was 3.5 +/- .9 umol/mg P for the control group and 4 +/- 0.2, 8.2 +/- 0.9 and 11 +/- 1 for 1, 2 or 8 weeks of CsA treatment, respectively. These findings appear to indicate that the development of significant gingival changes induced by CsA is related to new-onset of diabetes mellitus during the immunosuppressive treatment. PMID- 21638967 TI - The antimicrobial effect of iodine-potassium iodide after cleaning and shaping procedures in mesial root canals of mandibular molars. AB - The aim of this study was to evaluate the antibacterial activity of 2% iodine potassium iodide (IKI) used as a final rinse after the cleaning and shaping procedures in mesial root canals of mandibular molars infected with Enterococcus faecalis. Seventy two mandibular first molars were used. The root canals were infected with Enterococcus faecalisfor 30 days. After the infection procedures, the root canals were cleaned and shaped by using the Pro Taper rotary system and manual files. The teeth were randomly assigned to four experimental groups (N = 18). In group 1, the root canals were irrigated with sterile distilled water (control). In group 2, the root canals were irrigated with 1% Sodium hypochlorite (NaOCl) during instrumentation. In group 3, root canals were irrigated with 1% NaOCl during instrumentation and a five minute final irrigation using 2% IKI. In group 4, the root canals were irrigated with 1% NaOCl during instrumentation and a 15 minutes final irrigation with 2% IKI. Bacteria colony-forming units (CFU) from root canals were semi-quantified and the presence of negative cultures among the groups was compared using Fisher's test (p < 0.05). The order of effectiveness was: 1% NaOCI plus 2% IKl for 15 minutes (95%), 1% NaOCl plus 2% IKl for 5 minutes (44%), 1% NaOCl (17%) and sterile distilled water (0%). Fisher's exact test showed a significant difference among the groups (p < 0.05). It was concluded that under in vitro conditions, IKI was able to eliminate the Enterococcus faecalis from infected dentin significantly in a 15-minute time frame after the cleaning and shaping procedures. PMID- 21638968 TI - Physicochemical salivary properties, Lactobacillus, mutans streptococci counts and early childhood caries in preschool children of Colombia. AB - Dental caries is a multifactorial disease, in which saliva has an important role. Saliva properties affect the growth of cariogenic microorganisms, so variations below threshold levels are considered risk factors for the development of dental caries. Salivary tests and microbiological counts of cariogenic bacteria are often used for clinical purposes in order to reveal problems that could explain caries development and have been evaluated in multiple studies, but these studies have mainly been carried out on adults. Thus, the aim of this study was to identify physicochemical salivary properties, mutans streptococci and Lactobacillus counts among preschool children, and their relationship with ECC in a cross-sectional study in Medellin, Colombia. Caries was determined using ICDAS criteria for a sample of 201 preschool children. Stimulated saliva samples were processed for bacterial detection, mean flow rate, pH and buffer capacity calculation. Risk variables for ECC were assessed individually and adjusted using a binary logistic regression model. The results showed that 71.14% of study subjects had ECC. Flow rate, final pH and Buffer capacity increased with age, but none of them were related to the presence of ECC. Although OR analysis detected correlations between ECC, gender age, dental eruption stage and Lactobacillus presence, binary logistic regression analysis only showed gender and Lactobacillus as strongly associated with ECC. It can be concluded that gender and Lactobacillus presence are key variables for the presence of ECC in these children; whilst age and dental eruption stage play a minor role. Further studies are required to clarify the role of mutans streptococci counts in this population and to establish normal salivary parameters as well as threshold levels for hyprosalivation in preschool children, as results showed differences with normal parameters used for adults. PMID- 21638969 TI - Effect of bleaching agent on dental ceramics roughness. AB - The aim of this study was to assess the effect of bleaching agents (10% and 16% carbamide peroxide) on the roughness of two dental ceramics in vitro, and to analyze the surface by scanning electronic microscopy (SEM). Two bleaching agents (10% and 16%/Whiteness, FGM Gel) and two microparticle feldspathic ceramics (Vita VM7 and Vita VM13) were used. Forty disks of Vita VM7 and Vita VM13 ceramic were manufactured, measuring 4 mm in diameter and 4 mm high, in accordance with the manufacturers' recommendations, and were divided into 4 groups (n = 10): (1) VM7 + Whiteness 10%; (2) VM7 + Whiteness 16%; (3) VM13 + Whiteness 10%; (4) VM13 + Whiteness 16%. The bleaching agent was applied for 8 hours a day for 15 days and during the intervals the test specimens were stored in distilled water at 37 degrees C. The roughness (Ra) of the test specimens was evaluated before and after exposure to the bleaching agents using a laser roughness meter and the topographic description was analyzed by SEM. The statistical analysis of roughness data showed significant differences in the VM7 groups, using paired t test, p = 0.05 (VM7 + Whiteness 10%: p = 0.002; VM7 + Whiteness 16%: p = 0.001) and two-sample t-test (VM7 p = 0.047), and no significant difference was found among VM13 groups. The qualitative SEM analysis showed different degrees of surface changes. The results suggest that the roughness of the tested ceramic surfaces increased after exposure to the bleaching agents. PMID- 21638970 TI - Evaluation of vascular endothelial growth factor (VEGF) in interradicular bone marrow in olpadronate treated animals. AB - Vascular endothelial growth factor (VEGF) is a protein that increases vascular permeability and induces the proliferation, migration and survival of endothelial cells. Bisphosphonates (BPs) are antiresorptive drugs that are widely used in the treatment of bone metabolism diseases and bone metastases. Since 2003, cases of bisphosphonate-related osteonecrosis of the jaw (BRONJ) have been reported. Few papers explain the mechanisms that induce BRONJ; some authors mention alterations in bone remodelling and a certain antiangiogenic effect of BPs. The aim of this study is to evaluate the expression of VEGF in bone marrow cells and the number of blood vessels and area occupied by them in animals treated with the BP sodium olpadronate (OPD). We used 16 Wistar rats, 60 days old, divided into two groups, experimental (OPD) and control. The OPD group received 0.3 mg/kg/week intraperitoneal OPD for 5 weeks. The control group received an equivalent intraperitoneal volume of physiological saline solution. After euthanasia, hemimandibles were processed and mesio-distal histological sections of the first molar were prepared. Sections were stained with hematoxylin-eosin (HE), immunohistochemical detection of VEGF was performed (sc-7269) and the following histomorphometric parameters were evaluated: In HE-stained sections--number of blood vessels per sq. mm. and percentage (%) of area occupied by blood vessels in relation to total area evaluated; in sections with immunohistochemical detection of VEGF--number of VEGF+ bone marrow cells per sq. mm. Data underwent statistical analysis. Number of blood vessels/mm2 was significantly lower in the OPD group (OPD: 92 +/- 16; CONTROL: 140 +/- 31; p < 0.05) and % vascular area/total area evaluated showed no significant difference (OPD: 15.6 +/- 6.1; CONTROL: 10.2 +/- 4.2). Number of VEGF+ cells/mm2 was lower in the OPD group than in the control group, with statistically significant differences (OPD: 7804.8 +/- 597; CONTROL: 13187.6 +/- 894; p < .001). The results of this study suggest that monosodium olpadronate has an antiangiogenic effect. Further studies are needed to reveal its potential as an antitumor agent and its connection with the onset of BRONJ. PMID- 21638971 TI - Top HIS vendors by revenue. PMID- 21638972 TI - A new path for health care. PMID- 21638973 TI - To scan or not to scan. PMID- 21638974 TI - Using information technology market differentiator. PMID- 21638975 TI - Overcoming ICD-10 hurdles. PMID- 21638976 TI - Home monitor. Interview by Elizabeth Gardner. PMID- 21638977 TI - Prevention of disability in leprosy: the different levels. AB - Prevention of disability in people affected by leprosy is primarily seen as prevention and management of impairments secondary to nerve function impairment. This article describes four different levels at which appropriate interventions may lead to the overall prevention of disability. These are--prevention of disease, timely diagnosis and adequate treatment of the disease, early recognition and adequate treatment of nerve function impairment and finally, prevention and treatment of secondary impairments due to nerve function loss. PMID- 21638978 TI - An attempt towards prevention and management of disabilities and deformities in leprosy. AB - History of prevention of deformities is practically as old as the appearance of the deformities themselves, unfortunately without much understanding to start with. In medieval era and even earlier, leprosy and deformities were treated synonymously and the disease's infectivity too was closely associated with appearance of deformities. Hence, to reduce chances of deformities caused by leprosy in healthy population, the patients having deformities were driven away from the society presuming that only deformed patients spread the disease. Unfortunately, it never worked. However, in later period, factors behind the deformities and disabilities were recognized and understood. These are basically limited to involvement of peripheral nerves and their proper management (medical treatment, surgical interventions, physiotherapy, ergonomics and counseling) by one rule of thumb i.e. early, timely and adequately. PMID- 21638979 TI - Nerve conduction studies in paucibacillary and multibacillary leprosy: a comparative evaluation. AB - Involvement of peripheral nerves in patients with leprosy results in sensory, motor and autonomic dysfunctions along with deformities and disability. Pattern of nerve involvement is different for different forms of leprosy. In this study, we evaluated and compared the nerve conduction parameters of paucibacillary leprosy with that of multibacillary leprosy. In this study, 40 consecutive patients of leprosy (19 cases of paucibacillary and 21 cases of multibacillary leprosy) were included. Nerve conduction studies were performed according to the standard procedure described in the manual of the machine. We observed that patients with multibacillary leprosy had significantly more severe changes in nerve conduction parameters as compared to that of paucibacillary leprosy. In paucibacillary leprosy, the dominant pattern of nerve involvement was that of mononeuropathy, however, in 6 paucibacillary cases the nerve involvement was in form of mononeuritis multiplex. Electrophysiological assessment also revealed involvement of clinically uninvolved nerves. Nerve conduction parameters were suggestive of mixed axonal as well as demyelination of the peripheral nerves. PMID- 21638980 TI - Leprosy among tribal population of Chhattisgarh state, India. AB - Chhattisgarh state is still endemic for leprosy and has a large tribal population. During 2003-2009, a total of 1530 untreated leprosy cases reported to the Leprosy Mission Referral Hospital in Champa, of which 151(9%) were classified as belonging to the scheduled tribes. The characteristics of these new tribal patients are described and compared with other patients and to the demographics of the tribals in the general population of the State. While tribals were accessing the leprosy services similar to the other social groups, the delay in reporting, high BI and other features pose more serious problems in the transmission of leprosy and in management of complications due to the harsh environment and occupational patterns of the tribals. Appropriate strategies and more community based approaches will be necessary if these groups are also targeted for eradication of leprosy. PMID- 21638981 TI - Response of thickened nerve trunks and skin lesions of leprosy patients to MDT. AB - This paper indicates the responses of thickened nerve trunks in leprosy patients to MDT. Out of 1625 cases, 557 (34.2%) cases had thickened nerve trunks at the time of registration. From these cases, 175 (31.4%) were randomly selected and re examined by personal visit about 5 years after RFT. Follow-up showed persistent thickening in 96 (54.8%) cases. Persistence of nerve thickening was higher in MB leprosy. Additional nerve thickening appeared in 8 (4.6%) cases. New disability developed in 6 (3.4%) cases after RFT but these persons did not come for check up voluntarily. Reaction occurred in 6 during follow-up. Both MB and PB considered together thickening continued in as high as 96 (54.8%) compared to persisting skin lesions in 24 (13.7%) cases. Persons with thickened nerve trunks require more counseling to report for check up at the earliest sign of nerve function deficit. PMID- 21638982 TI - Effects of environment and education on knowledge and attitude of nursing students towards leprosy. AB - The objective of this cross-sectional comparative study was to find the effects of environment and education on knowledge and attitude of nursing students towards leprosy. Data were collected, using a pretested questionnaire, from the first year and third year students of a School of Nursing attached to a leprosy specialty hospital and also from a comparable School of Nursing attached to a general hospital. The results showed that trainees acquired more knowledge on leprosy during training in both schools of nursing. However, those trained in leprosy hospital environment had higher knowledge and attitude scores than those trained in general hospital environment. The attitude of the trainees attached to leprosy hospital was favourable even before they had formal training in leprosy. Those trained in the general hospital showed more favourable attitude after training compared to before training. School of Nursing attached to leprosy hospital provided an atmosphere conducive to learning and understanding more about leprosy. The trainees retained what was learnt because of regular association with patients affected by leprosy. For employment in hospital or community based services or research related to leprosy, nurses trained in a leprosy hospital would have added value of knowledge and attitude. PMID- 21638983 TI - Can the skin smear examination in NLEP be reconsidered? AB - Slit and skin smear examination is an important laboratory test which was made optional in the NLEP. In a laboratory managed by LEPRA Society, this test was provided to voluntary reporting cases and those referred by the general health service institutions of the area. Total cases tested were 1180, out of which 152 (12.9%) were positive. Of the 152 smear reports, 121 were from Koraput district and constitute the sample for this study. Regarding the spread of site wise highest BI, 56% cases had BI of 3.1+ to 6+. Of the 121 cases, 112 (92.6%) cases completed 12 doses of MDT. Seventy patients, from whom detailed information was collected by personal contact, responded well to MDT resulting in regression of lesions. Out of them, 46 cases had repeat smear showing negative smear in 39% and decline of 81 in the rest of cases. There are 16 contact cases with 2 family clusters. A rough block-wise mapping showed 4 strata with a dense clustering in the urban area where the laboratory facility is available. The probable reasons for uneven distribution of cases and causes for late diagnosis were discussed. The study reflects the leprosy problem in urban pockets which high infection pool. Together these two factors remain unexposed due to winding up of smear labs and pose a potential threat to elimination. The paper also emphasized on the reasons for past failure of the test along with suggestions to revive this facility with minor modifications. PMID- 21638984 TI - Satisfaction of leprosy patients on integrated MDT services: as evaluated by Bargarh Integrated Community Health Project (BICHP). AB - Functional integration can be viewed successful if general health system (GHS) delivers services to the satisfaction of leprosy patients. The study elicits responses from 307 leprosy cured persons of 3 blocks and one town covering a population of about 3,50,000 in district Bargarh, Orissa. 229 (74.6%) MB and 78 (25.4%) cured PB cases were the respondents. About 80% patients diagnosed and received treatment within half an hour, 89.9% respondents spent adequate time with providers for clarifying different aspects of leprosy, 85.0% felt the discussion very useful. About 94% had comfortable time at the health institutions. About 97% of persons were regular and completed FDT in time while 25.0% experienced some discrimination mostly at the health institutions. Trunk nerves were involved in 57% cases at detection which subsided during treatment in 33% of cases. Skin patches disappeared in 74.6% and became inactive in the rest. Majority (90.2%) were convinced about cure by MDT. Clients' satisfaction in terms of quality treatment and cordial patient provider relation in GHS should be a strong indicator for successful integration. PMID- 21638985 TI - Predictors of adverse outcome in a diabetic population following acute coronary syndromes. AB - INTRODUCTION: People with diabetes are at increased risk for heart failure (HF), major adverse cardiovascular events (MACE) and death following acute coronary syndromes (ACS). It is important to recognize the most powerful predictors of these events after an ACS as early as possible, in order to address them more aggressively. This is particularly important considering that various studies have shown that this population is undertreated in the setting of ACS. OBJECTIVES: To characterize a diabetic population presenting with ACS and to determine independent predictors of HF, MACE and mortality on follow-up. METHODS: This was a longitudinal, observational, retrospective study including 471 consecutive diabetic patients, both previously known and newly diagnosed, hospitalized for ACS in a single center between May 2004 and December 2006. A mean 12-month follow-up was conducted. Cox regression analysis was used to determine the independent predictors of HF, MACE and mortality on follow-up, divided into different periods--1 month, 6 months and 1 year. RESULTS: Of the overall diabetic population, 67.3% were male and mean age was 69 +/- 11 years. Mean glomerular filtration rate (GFR) was 62 +/- 22 ml/min and mean left ventricular ejection traction (LVEF) was 50%. diagnosis on admission was ST elevation myocardial infarction (STEMI) in 31.3%, non-ST elevation myocardial infarction (NSTEMI) in 50.1%, unstable angina (UA) in 14.3% and ACS with left bundle branch block or pacemaker in 4.2%. Cardiac catheterization was performed in 55.6% of the patients during the index hospitalization. Mortality during hospitalization and at 1 year was 6.4% and 10.4%, respectively. The one-year MACE rate was 20.4% and hospitalization for HF occurred in 10.1% of the patients. The independent predictors of HF at 1 year were blood glucose on admission > 184.5 mg/dl, GFR < 63.8 ml/min, LVEF < 46.5% and NSTEMI, while predictors of mortality were LVEF < 40.5% and Killip class on admission > I. Blood glucose on admission > 130.5 mg/dl and LVEF < 49.5% were independent predictors of MACE, whereas cardiac catheterization was a protective factor. CONCLUSION: Following ACS diabetic patients have high rates of mortality, HF and MACE. The low rate of invasive strategy may contribute to this situation. HF during hospitalization, whether by low LVEF or Killip class > I, and higher blood glucose on admission were powerful predictors of poorer outcome. Moreover, the use of recommended cardiovascular agents and procedures were protective factors. These findings suggest that diabetic patients should not be excluded from recommended cardiovascular interventions. Efforts should be made to identify these high-risk patients as early as possible in order to manage them carefully and aggressively to improve their poor prognosis. PMID- 21638986 TI - Diabetes, ventricular dysfunction and acute coronary syndrome: an explosive triad. PMID- 21638987 TI - Benefits of cardiac resynchronization therapy in "very dilated cardiomyopathy". AB - INTRODUCTION: Recent clinical trials have studied parameters that could predict response to cardiac resynchronization therapy (CRT) in patients with advanced heart failure. Left ventricular end-diastolic dimension (LVEDD) is regarded as a possible predictor of response to CRT. OBJECTIVE: To study the response to CRT in patients with very dilated cardiomyopathy, i.e. those at a more advanced stage of the pathology, analyzing both the responder rate and reverse remodeling in two groups of patients classified according to LVEDD. METHODS: We performed a retrospective analysis of 71 patients who underwent CRT (aged 62 +/- 11 years; 65% male; 93% in NYHA functional class > or = III; 31% with ischemic cardiomyopathy; left ventricular ejection fraction [LVEF] 25.6 +/- 6.8%; 32% in atrial fibrillation; QRS 176 +/- 31 ms). Twenty-two (31%) patients with LVEDD > or = 45 mm/m2 (49.2 +/- 3.5 mm/m2) were considered to have very dilated cardiomyopathy (Group A) and 49 patients had LVEDD > 37 mm/m2 and < 45 mm/m2 (39.4 +/- 3.8 mm/m2) (Group B). All patients were assessed by two-dimensional echocardiography at baseline and six months after CRT. The following parameters were analyzed: NYHA functional class, LVEF and LVEDD. Responders were defined clinically (improvement of > or = 1 NYHA class) and by echocardiography, with a minimum 15% increase over baseline LVEF combined with a reduction in LVEDD (reverse remodeling). RESULTS: There were no significant differences in baseline demographic characteristics between the two groups. At six-month followup, we observed an improvement in LVEF (delta 8.5 +/- 11.8%) and a reduction in LVEDD (delta 3.7 +/- 6.8 mm/m2), with fifty-seven (79%) patients being classified as clinical responders. The percentage of patients with reverse remodeling was similar in both groups (64% vs. 73%, p = NS), as were percentages of improved LVEF (delta 6.3 +/- 11% vs. delta 9.6 +/- 12%; p = NS) and decreased LVEDD (delta 3.7 +/- 5.5 mm/m2 vs. delta 3.7 +/- 7.4 mm/m2; p = NS). We found a higher percentage of clinical responders in patients with very dilated cardiomyopathy (96% vs. 72%, p < 0.05). CONCLUSION: In this study, a significant number of responders showed reverse remodeling after CRT. Although a higher percentage of patients with very dilated cardiomyopathy showed improvement in functional class, the extent of reverse remodeling was similar in both groups. PMID- 21638988 TI - Left ventricular noncompaction: analysis of a pediatric population. AB - INTRODUCTION: Left ventricular noncompaction (LVNC) is a rare and potentially progressive cardiomyopathy, characterized by the persistence of multiple trabeculations and deep intratrabecular recesses in the ventricular myocardium. Although two-dimensional and color Doppler echocardiography are the most useful diagnostic modalities, cardiac magnetic resonance imaging has proved to have high sensitivity and specificity in the diagnosis of this anomaly. OBJECTIVE: To characterize the clinical and imaging features of LVNC in a pediatric population and to assess their evolution. METHODS AND RESULTS: We performed a retrospective chart review of five pediatric patients with LVNC, followed at Coimbra Pediatric Hospital between January 1999 and December 2007. Median age at presentation was five months (ranging from one day to 13 years), and they were mainly male (1.5:1). Two of the children had a family history of sudden death. In one case the clinical presentation was cardiac arrest due to ventricular fibrillation and in three others, congestive cardiac failure. None of the five cases had associated congenital cardiac anomalies. Involvement of the ventricular apical region was found in all cases. Four children additionally had ventricular dysfunction which improved with diuretic and vasodilator therapy. Mean follow-up was 34 months, ranging from six months to seven years. In one case a change in the morphological phenotype was noted, from a dilated to a hypertrophic form. In this case and in the child's father a mutation in the MYBPC3 gene was identified, which is associated with hypertrophic cardiomyopathy. No thromboembolic phenomena or deaths occurred during the study period. CONCLUSION: In the pediatric population, congestive cardiac failure is the most common clinical presentation of LVNC, which can coexist with other cardiomyopathies, particularly dilated and hypertrophic forms. The sample presented in this analysis is statistically non significant due to its limited size and the authors highlight the need for larger prospective studies in the pediatric population in order to clarify this disease and its diagnostic criteria. PMID- 21638989 TI - Is the recommendation to walk sufficient to achieve the levels of physical activity recommended to obtain cardiovascular benefits? AB - The aim of the present study was to assess whether subjects with cardiovascular risk factors who are referred to participate, without supervision, in walking programs, or do so voluntarily, achieve the levels of daily physical activity recommended to obtai cardiovascular benefits. To this end, an observational descriptive cross-sectional study was conducted. The sample was composed of 23 subjects (age: 43.9 +/- 14.5 years) with at least one modifiable cardiovascular risk factor, who took part in a walking program following referral from their general practitioner or by self-referral. The intensity and duration of physical activity were assessed by accelerometry. The subjects wore an accelerometer on a belt in the region of the right anterior iliac crest. The accelerometer data are reported as minutes spent in physical activity of light, moderate, vigorous and very vigorous intensity. All subjects in our sample walked at the levels of moderate-intensity physical activity recommended to obtain cardiovascular benefits, performing 40.01 +/- 6.2 minutes of moderate-intensity physical activity and only 5.7 +/- 3.0 minutes of light-intensity physical activity in the walking session, which lasted a mean of 45.8 +/- 7.0 minutes. The results of the present study indicate that the simple referral of subjects with cardiovascular risk factors to walk is sufficient to achieve the levels of moderate-intensity physical activity recommended to obtain cardiovascular benefits. PMID- 21638990 TI - Noncompaction of the ventricular myocardium: characterization and follow-up of an affected population. AB - Noncompaction of the ventricular myocardium (NVM) is a rare congenital disease caused by an arrest in normal myocardial embryogenesis, leading to persistence of numerous prominent trabeculations which communicate with the left ventricle. It was first described as a congenital condition affecting children, but several cases have been reported of late presentation, and recent studies suggest it may be underdiagnosed. The main clinical manifestations are congestive heart failure, arrhythmias (supraventricular or ventricular) and systemic embolism. We describe a series of twenty patients, focusing on clinical history, echocardiography and follow-up. PMID- 21638991 TI - Very late coronary stent thrombosis after discontinuation of antiplatelet therapy. AB - Percutaneous transluminal coronary angioplasty (PTCA) with stent placement is widely used to achieve myocardial revascularization in patients with symptomatic ischemic heart disease and significant coronary artery stenosis. Drug-eluting stents are used in most patients undergoing percutaneous angioplasty. Stent thrombosis is an uncommon but serious complication, manifested mostly by sudden death or acute ST-elevation myocardial infarction. The authors report the case of a 68-year-old patient with acute anterior ST-elevation myocardial infarction. Five years previously, she had had a similar presentation and underwent primary angioplasty of the left anterior descending artery with implantation of a drug eluting stent. The patient was discharged under antithrombotic therapy. She discontinued antiplatelet therapy and two days later suffered an acute anterior myocardial infarction. Primary angioplasty revealed stent thrombosis. PMID- 21638992 TI - Something inside the heart: a myxoid liposarcoma with cardiac involvement. AB - A 70-year-old man presented with dry cough, dyspnea and fatigue. Physical examination revealed only a grade 3 heart murmur. The chest X-ray showed bilateral nodular opacities. A CT scan revealed several nodular masses in both lungs, pleura and abdomen, as well as a similar lesion inside the left ventricle. Echocardiography revealed not only the pedunculated nature of this lesion, but also infiltration of the ventricular wall. Biopsy showed it to be a myxoid liposarcoma. Since the disease was already widespread, and given the characteristics of the intraventricular mass, only palliative measures were prescribed. PMID- 21638993 TI - National Registry on Cardiac Electrophysiology 2009. AB - The authors analyze the number and type of electrophysiologic procedures (diagnostic and ablation, and implantation of defibrillators and biventricular pacemakers with defibrillator backup) that were performed during 2009 in all Portuguese electrophysiology centers. A total of 2669 diagnostic electrophysiologic procedures were performed during the year, of which 62% were followed by ablation, amounting to 1668 ablations. The authors describe the type of ablations performed by the various centers, and analyze the distribution and type of ablation procedures in the light of recent data from published international studies. A total of 1089 ICDs were implanted during 2009, and the authors also describe the type of devices employed and comment on these data bearing in mind recent published data. PMID- 21638994 TI - Aortic valve balloon dilatation with rapid right ventricular pacing. PMID- 21638995 TI - Asymptomatic cor triatriatum in an adult--diagnosis by cardiac CT angiography. PMID- 21638996 TI - Prostate cancer incidence and mortality in selected countries of Central Europe. AB - BACKGROUNDS: This paper analyzes the incidence and mortality of prostate cancer in the Slovak (SR) and Czech (CR) Republics (as Central European countries with population-based cancer registries) before and after the introduction of PSA testing, the possible reasons for any differences disclosed, and compares the results with selected regions and countries around the world. MATERIAL, METHODS AND RESULTS: In SR, the age-adjusted incidence of prostate cancer rose from 14.6/100,000 in 1968 to 36.2/100,000 in 2005. The estimated annual increase of incidence from 1968 to 1991 (before nation-wide PSA testing) was 0.421 and 1991 2003 it reached 0.941. The mortality rates rose from 7.3/100,000 in 1968 to 14.9/100,000 in 2005. The increase in incidence occurred faster in CR than in SR, from 15.8/100,000 in 1977 to 59.5/100,000 in 2005. The estimated annual increase of incidence in CR in 1977-1991 was 0.581,while in 1991-2003 it reached 1.981. Before 1991, mortality rose more sharply in CR than in SR while after the introduction of PSA testing mortality stabilized more quickly in the CR than in SR. In SR a significant reduction of mortality was observed after 2002 and is probably affected by more factors than those associated with the increase in PSA testing. CONCLUSION: The difference in the incidence and mortality of prostate cancer in SR and in CR results from a difference in the intensity of PSA testing as well as from the earlier introduction of effective treatment in CR. PMID- 21638997 TI - Angiomyofibroblastoma of the cervix uteri: a case report. AB - BACKGROUNDS: Angiomyofibroblastoma (AMFB) is a rare histopathologic finding of the female lower genital tract. This tumor belongs to the group of mesenchymal tumors. Mesenchymal neoplasms of the modified genital skin and mucosa are uncommon. The majority of these lesions are seen in females and, collectively, they form a family of vulvovaginal soft tissue tumors. This family includes fibroepithelial stromal polyps, angiomyofibroblastoma, cellular angiofibroma, aggressive angiomyxoma, vaginocervical myofibroblastoma, vulvar leiomyomatosis, and other smooth muscle tumors. Angiomyofibroblastoma is a benign tumor, histologically very similar to pelvic aggressive angiomyxoma (AMM), a distinctive, locally infiltrative but non-metastasizing mesenchymal neoplasm with a tendency to occur in the female pelvic and perineal regions. CASE: 44 years old woman with angiomyofibroblastoma of cervix uteri. CONCLUSION: A recognition of this entity is important to avoid misdiagnosis of other angiomyxoid neoplasms. Furthermore, unlike other, more aggressive, mesenchymal tumors of the lower genital tract, AMFB shows benign behaviour. PMID- 21638999 TI - Press the button? PMID- 21638998 TI - Phase 0 clinical trials will overcome stagnation of anticancer drug development? AB - Recent guidance from the US Food and Drug Administration supports the conduct of a new type of exploratory clinical trials, commonly called phase 0 clinical trials, on the development of innovative anticancer agents, particularly targeted agents. Phase 0 clinical trials are controversial mainly because of the lack of clinical benefit to the participant patients. However, it was recognized that Phase 0 clinical trials can provide a platform to assessing the biological effects on the targets in tumoral human samples, evaluate biomarkers for drug effects and to generate essential human pharmacokinetics and pharmacodynamics data earlier in the drug development. It is expected that such trials will became a routine part of early-phase oncological drug development in the future. PMID- 21639000 TI - The anarchy of antibiotic resistance: mechanisms of bacterial resistance. PMID- 21639001 TI - Industry execs 'go with the flow'--flow-cytometry trends, that is. PMID- 21639002 TI - Phlebotomy formulas. PMID- 21639003 TI - Five rules that can build effective training objectives and plans. PMID- 21639004 TI - Addressing management issues. PMID- 21639005 TI - CMS ranks labs as 'moderate' risk. PMID- 21639006 TI - Blood cultures--in a nutshell. PMID- 21639007 TI - Five trends to watch. PMID- 21639008 TI - Unplugged: the code of life. PMID- 21639009 TI - Time to prepare for dengue testing? Florida Bureau of Labs is already on the job. PMID- 21639010 TI - Employee engagement vital to ISO 15189 success. PMID- 21639011 TI - Schwartz combines pathology with technology at Aperio. PMID- 21639012 TI - Factor structure and psychometric properties of the center for epidemiologic studies depression scale (CES-D) in older populations with and without cognitive impairment. AB - The CES-D is widely used for the assessment of depressive symptoms in the adult population. However, few studies have been performed to assess the utility of this scale in an older population with cognitive impairment. The factor structure of the Spanish version of the CES-D was examined in an observational, cross sectional study in 623 older adults (M = 72.74 years; SD = 7.7). The validity of the scale was determined in two samples of older adults, one comprising 162 participants with cognitive impairment (M = 76.73 years; SD = 8.1) and one with 58 participants without cognitive impairment (M = 74.64 years; SD = 9.0). The results confirm previous results of a four factor structure. With regard to the validity of the scale, in the group with cognitive impairment the area under the ROC curve is 0.84 (95% CI: 0.77-0.89) and the cut-off point for possible depression is 13, while in the group without cognitive impairment the area is 0.90 (95% CI: 0.79-0.96) and the optimal cut-off point is 28. These results show that the Spanish version of the CES-D is a valid instrument for the identification of depression in older adults with and without cognitive impairment. PMID- 21639013 TI - Development of the parent adult relationship questionnaire (PARQ). AB - Relationships between adults and parents are characterized by positive and negative qualities. Existing measures of relationship qualities and ambivalence often rely on items that are unbalanced in number of positive and negative items, emotional tone, or missing negative items completely. Three studies established validity (construct, convergent, discriminant) and reliability (internal consistency, test-retest) for the Parent Adult Relationship Quality (PARQ) scale. Study 1 found high internal consistency and test-retest correlations among undergraduates. Study 2 found associations with measures of positive relationship quality and high test-retest correlations using a sample of parents. Study 3 established convergent and discriminant validity with other measures of relationship quality, including an observational measure. The PARQ is a succinct measure that captures positive and negative aspects of support and interactions in relationships between adults and their parents. PMID- 21639014 TI - The impact of perceived stress, social support, and home-based physical activity on mental health among older adults. AB - This study investigated how perceived stress, social support, and home-based physical activity affected older adults' fatigue, loneliness, and depression. We also explored whether social support and physical activity mediated the relationships between stress and mental health problems. The data of 163 older participants were analyzed in this study. Structural equation modeling using LISREL 8.71 was performed to assess the effects of stress, support, and physical activity on mental health. The findings indicate that perceived stress predicted higher levels of depression, social support predicted lower levels of loneliness and fatigue, and physical activity predicted lower levels of fatigue among older adults. Social support and physical activity mediated the relationships between stress and mental health, except depression. In conclusion, the relative impacts of perceived stress, social support, and physical activity on types of mental health (e.g., fatigue, loneliness, and depression) were different. Furthermore, stress had direct and indirect effects on each construct of mental health (e.g., fatigue, loneliness, and depression). PMID- 21639015 TI - Proactive coping in community-dwelling older Australians. AB - This article reports on a study of older community-dwelling Australian adults which aimed to test whether a relatively unexplored construct, proactive coping, could have a role in purpose in life, personal growth, and life satisfaction. A total of 109 women and 115 men (Mean age = 75.04 yrs, SD = 6.66) completed a questionnaire containing Greenglass et al.'s (1999) Proactive Coping Inventory, the Purpose in Life and Personal Growth subscales of Ryff's (1989) Psychological Well-being Scales, and the Satisfaction with Life Scale (Diener et al., 1985). The results of hierarchical multiple regression analyses indicated that proactive coping was a highly significant predictor of all three measures of well-being, after controlling for age and health. The effect on personal growth and purpose in life was particularly strong. Proactive coping does seem to be an important variable in the psychological well-being of older adults. However, the correlational nature of the design, and the likelihood of some conceptual overlap between the well-being variables, suggest that these inferences can only be tentative. Designing psychological interventions to improve proactive coping may help to improve quality of life at older ages. There is a need for experimental research to further explore the causal influence of proactive coping and for further theoretical work to determine the exact nature of proactive coping. PMID- 21639016 TI - Lebron v. Gottlieb Memorial Hospital: capping medical practice reform in Illinois. AB - On February 4, 2010 the Illinois Supreme Court struck down Illinois' medical practice reform bill, P.A. 94-677, when it decided Lebron v. Gottlieb Memorial Hospital. Although the court only considered the unconstitutionality of the cap on non-economic damages contained in that bill, an inseverability provision resulted in the invalidation of all of its provisions. The end result of the Lebron decision extends much further than the striking of the cap on non-economic damages. It affects such areas as medical liability insurance law, physician discipline, public disclosure of information, the admissibility of physician statements into evidence, and expert witness standards. The Lebron holding has raised a significant obstacle to the implementation of a wide range of legislative measures intended to improve both the delivery and quality of healthcare services in Illinois. This article explains the impact of Lebron in these collateral, but important, areas of the law. PMID- 21639017 TI - The Stark Law in retrospect. AB - Considering the ultimate goals of preventing the over-utilization of medical services and protecting the Medicare program, are the numerous phases of the Stark Law and their concomitant regulations effective; or, conversely, has the legislation served to impede entrepreneurialism among physicians to the detriment of innovations and better integration in the delivery of medical treatment? This article endeavors to answer the above question through an analysis of the policy goals behind the legislation; the evolution of its regulations; its effect on competitiveness in the field of medicine; and the ethical considerations implicated by the issue of physician self-referral. It further offers some proposals which attempt to address the problem of physician self-referral abuse while at the same time reducing the complexity and breadth of the Stark law and its regulations. The article concludes by noting that to truly change the practice of inappropriate self-referral as well as the culture of over utilization, it is necessary not only to target specific relationships and practices prone to abuse, but to realign the financial incentives created by our current payment mechanisms as well. PMID- 21639018 TI - The false claims act and the eroding scienter in healthcare fraud litigation. AB - This article addresses the federal government's expansive methods in tackling healthcare fraud, particularly in misapplying the False Claims Act. Although tasked with the obligation to curtail the fraudulent submission of Medicare & Medicaid claims, the U.S. government must rein in the current trend to utilize the False Claims Act against smaller medical providers. As the Act's original focus has ebbed in significance, the government has increasingly applied the False Claims Act to circumstances that do not evince actual fraud. In doing so, federal courts have effectively eroded the statute's critical scienter requirement. The federal common-law doctrines of "payment by mistake" and "unjust enrichment" adequately address the payment of non-fraudulent, albeit false, Medicare & Medicaid claims. Yet the federal government pursues these appropriate remedies only rarely and in the alternative, essentially when the government fails under the False Claims Act. Thus, this article argues for reform, calling for a clearer delineation between remedial and punitive measures. In cases involving smaller medical providers, courts should strictly limit the False Claims Act to those instances where fraud is clearly manifest. PMID- 21639019 TI - Squeezing the middleman: ending underhanded dealing in the pharmacy benefit management industry through regulation. AB - Pharmacy benefit management (PBM) companies are the middlemen of the pharmaceutical industry, designing plans for sponsors and insurers and pushing the products of manufacturers. Their unique position can often create conflicts of interest, which has been the basis of much litigation. This article reviews the structure of the PBM industry and analyzes concerns arising from its handling of prescription drug pricing, manufacturer rebates and discounts, and mail order pharmacies. After surveying several legislative proposals, it concludes with a comprehensive outline for legislation to eliminate underhanded dealing in the industry and lower the cost of prescription drugs. PMID- 21639020 TI - Is state power to protect health compatible with substantive due process rights? AB - Public health laws may mandate drastic limitations on individual liberty, such as forced medication and quarantine. This results in a tension between public health laws and guarantees of liberty such as the Due Process Clauses of the Fifth and Fourteenth Amendments to the United States Constitution. The Supreme Court has resolved this tension in favor of one or the other of these legal principles, depending on the facts and issues involved. Nevertheless, Supreme Court jurisprudence is internally consistent. The Court has applied a level of scrutiny that, while rigorous, is more flexible than strict scrutiny. I denote this as "enhanced public health scrutiny." Applying this scrutiny, the Court will uphold public health legislation if it protects an inchoate class of people who may not yet be identifiable, who will incur a specific disease or injury absent the law, but who will not experience this disease or injury if the law is enforced. If this doctrine were explicit, it would constitute a clear guideline to courts seeking to balance health and liberty concerns. This guideline would be consistent with current case law, and would not impact on law affecting reproductive liberty. PMID- 21639021 TI - The importance of good data: how The National Sample Survey of Registered Nurses has been used to improve knowledge and policy. AB - In 1977, the federal government launched the nation's largest and most significant program to collect data on the registered nurse (RN) workforce of the United States-the National Sample Survey of Registered Nurses (NSSRN). This survey is conducted by the U.S. Health Resources and Services Administration, first in 1977 and then every 4 years since 1980. This article offers the history of the NSSRN and a review of the ways in which the NSSRN data have been used to examine education, demographics, employment, shortages, and other aspects of the RN workforce. The influence this body of research has had on policymaking is explored. Recommendations for future research are offered, in the hope that future waves of the NSSRN will continue to be used to their fullest potential. PMID- 21639022 TI - Job satisfaction and retention of military nurses: a review of the literature. AB - Job satisfaction is an extremely important concept that influences a nurse's decision to stay in an organization, as well as the cost of turnover and the nursing shortage. The purpose of this review is to identify published research studies that have assessed job satisfaction and retention (intent to stay) in military nurses serving in the Army, Navy, or Air Force. The available literature was searched from 1980 to 2010 and the review resulted in 21 studies. The majority of the studies used 2 descriptive correlational design and was specific to one particular service. The researchers reported several satisfiers such as strong sense of teamwork, favorable work environments, pay and benefits, promotional opportunities, leadership and management experiences offered to junior officers. One of the major dissatisfiers was the lack of support from leadership. Nurse researchers must expand the retention science with robust longitudinal interventional studies. Nurse researchers are well positioned to provide military nurse leaders with the best possible evidence to address issues and make decisions regarding nurse retention. PMID- 21639023 TI - Understanding the supply and distribution of registered nurses: where are the data and what can they tell us? AB - Health workforce researchers routinely conduct studies to determine whether a profession is currently in short supply and whether future shortages are likely. This is particularly important for registered nursing since the profession has experienced periodic shortages over the past three decades. Registered nurse (RN) forecast studies can be valuable in quantifying supply and demand gaps and identifying the most appropriate strategies to avert future shortages. In order to quantify RN supply/demand gaps, it is important to have accurate data on RNs, including the number of active RNs as well as their demographic, education, and practice characteristics, and work location(s). A lack of relevant and timely data on the nursing workforce is a significant barrier to identifying where nursing shortages exist, where they are most severe, and determining the factors that contribute to them. This lack of understanding impedes the development of effective health workforce programs and policies to mitigate shortages and the ability to evaluate these programs and policies for effectiveness. This study describes the national data sources available to nursing researchers to study the supply and distribution of the RN workforce and assesses the sources' strengths and limitations. This study also explores the potential for using state-level data for nursing workforce research. PMID- 21639024 TI - State policy and research initiatives focused on improving nursing workforce: an integrative literature review. AB - The purpose of this chapter is to examine to and synthesize nursing workforce research and policy initiatives at the state level. An integrative literature review was systematically conducted using Ganong's Stages of an Integrative Research Review (1987). Searches were limited to English-language publications in the years from 2000 to 2010 which focused on workforce issues in the United States. A total of 155 published articles were included in this review. Overall, the literature indicated that significant research is examining nursing workforce issues and that states are actively engaged in policy initiatives to address nursing workforce issues, particularly those related to a shortage of nurses and faculty. The findings also indicated a major disconnect between nursing workforce research and patient outcomes research. Recommendations include connecting research and policy links between nursing workforce research and patient outcomes research and creating clear correlations to system-level determinants of quality Additional implications for further research are provided which include the important role of nurse researchers in connecting nursing care and nurse staffing to processes and outcome measures, which demonstrate the financial impact upon health care. PMID- 21639025 TI - An integrative review of global nursing workforce issues. AB - Migration has been a way of life since the beginning of time, with migrants seeking other lands for personal and professional betterment. Today, in an era of globalization, trade agreements and technological advances, an increase in migration is inevitable. All professions have been affected, but the migration of health professionals, particularly nurses, has been the most dramatic. However, the migration of nurses across national and international borders comes with many challenges: systematic tracking of migration flows, harmonization of standards, recognition of professional credentials, fair and equitable distribution of the global health care workforce, and the effect of migration on the health care infrastructure of both source and destination countries. The international migration of nurses to address shortages in developed countries has, in some instances, left source countries with insufficient resources to address their own health care needs. The increasing complexity of health care delivery, aging of the population and the nursing workforce, and the escalating global demand for nurses create on-going challenges for policy makers. Strategically addressing global nursing workforce issues is paramount to sustaining the health of nations. PMID- 21639026 TI - Bullying, harassment, and horizontal violence in the nursing workforce: the state of the science. AB - In the complex health care workplace of nurses, intra/interprofessional ideals intersect with the expectations of patients, families, students, and coworkers in a context of managed care environments, academia, and other health care enterprises. Integral to quality assessment, management, and assurance is collegial and respectful communication. Decades of reported descriptive and anecdotal data on intra/inter professional and on client communication, describe the antithesis of these ideals. Specifically, increasing frequency and rates of persistent bullying, harassment, or horizontal violence (BHHV) have shown to yield detrimental effects on workplace satisfaction, workforce retention, and the psychological and physical health of nurses as well as implied effects on quality of patient care and risk of poor health outcomes. Persistent BHHV among nurses is a serious concern. In advancing the science of description and explanation to a level of prevention intervention, explanatory models from biology, developmental psychology, intra/interpersonal interactionism are described along with theoretical explanations for the prevalence of BHHV in nurse workplaces. Making the connection between explanatory models and creative solutions to address BHHV through multiple levels of behavioral influence such as individual, environmental, interpersonal, and cultural contexts is key to advancing the science of the relationship between professional behavior and client/family/community health care outcomes. PMID- 21639027 TI - Interdisciplinary teamwork and collaboration: an essential element of a positive practice environment. AB - Interdisciplinary collaboration is critical to excellence in patient care delivery. There is a growing consensus that the basic education for all clinical professionals should include the knowledge, skills, and attitudes required to effectively participate in interdisciplinary teams, and that health care organizations should continue this education in the practice setting. The authors examine the large and growing evidence base regarding interdisciplinary collaboration and teamwork and explore the relationship between interdisciplinary collaboration and patient, workforce, and organizational outcomes. Antecedents and attributes of the construct are presented, as well as structures, models, and programs that are being implemented by health care organizations and academic settings to facilitate and advance interdisciplinary collaboration in clinical practice. PMID- 21639028 TI - The health care work environment and adverse health and safety consequences for nurses. AB - Nurses' working conditions are inextricably linked to the quality of care that is provided to patients and patients' safety. These same working conditions are associated with health and safety outcomes for nurses and other health care providers. This chapter describes aspects of the nursing work environment that have been linked to hazards and adverse exposures for nurses, as well as the most common health and safety outcomes of nursing work. We include studies from 2000 to the present by nurse researchers, studies of nurses as subjects, and studies of workers under similar working conditions that could translate to nurses' work environment. We explore a number of work organization factors including shift work and extended work hours, safety climate and culture, teamwork, and communication. We also describe environmental hazards, including chemical hazards (e.g., waste anesthetics, hazardous drugs, cleaning compounds) and airborne and bloodborne pathogen exposure. Nurses' health and safety outcomes include physical (e.g., musculoskeletal disorders, gastrointestinal, slips, trips and falls, physical assault) and psychosocial outcomes (e.g., burnout, work-family conflict). Finally, we present recommendations for future research to further protect nurses and all health care workers from a range of hazardous working conditions. PMID- 21639029 TI - Magnetism and the nursing workforce. AB - The focus of this chapter is to highlight practice exemplars and research findings related to the five components of the new Magnet Model. A brief overview of the historical development and professional evolution of the American Nurses Credentialing Center (ANCC) Magnet Recognition Program is presented followed by a brief overview of the original fourteen forces of magnetism. Content related to empirical practice-based research framed under the components of transformational leadership; structural empowerment; exemplary professional practice; new knowledge, innovation, and improvement; and empirical outcomes is presented and discussed. The authors provide key findings from scholarly publications and describe how the findings contribute to the creation of work environments based on the tenets of magnetism. The chapter concludes with a brief over of the ANCC Pathway to Excellence Program. PMID- 21639030 TI - Standardized nursing languages: essential for the nursing workforce. AB - The evolution of standardized nursing languages (SNLs) has been occurring for more than four decades. The importance of this work continues to be acknowledged as an effective strategy to delineate professional nursing practice. In today's health care environment, the demand to deliver cost-effective, safe, quality patient care is an essential mandate embedded in all health reform policies. Communicating the contributions of professional nursing practice to other nurses, health providers, and other members of the health care team requires the articulation of nursing's focus of concern and responses to these concerns to improve patient outcomes. The visibility of the electronic health record (EHR) in practice settings has accelerated the need for nursing to communicate its practice within the structure of the electronic format. The integration of SNLs into the patient record offers nurses an opportunity to describe the focus of their practice through the identification of nursing diagnosis, interventions and outcomes (IOM, 2010). Continued development, testing, and refinement of SNLs offers nursing an accurate and reliable way to use data elements across populations and settings to communicate nursing practice, enable nursing administrators and leaders in health care to delineate needed resources, cost out nursing care with greater precision, and design new models of care that reflect nurse-patient ratios and patient acuity that are data driven (Pesut & Herman, 1998). The continued use of nursing languages and acceleration of nursing research using this data can provide the needed evidence to help link nursing knowledge to evidence-driven, cost-effective, quality outcomes that more accurately reflect nursing's impact on patient care as well as the health care system of which they are a part. The evaluation of research to support the development, use, and continued refinement of nursing language is critical to research and the transformation of patient care by nurses on a global level. PMID- 21639031 TI - Results of an integrative review of patient classification systems. AB - This chapter presents the findings of an integrative review of the literature to identify current practices related to patient classification systems (PCSs). We sought to determine if there was a "gold standard" PCS that could be adopted or adapted for use by nurse leaders in practice. Sixty-three articles reporting studies related to PCS, Patient Acuity Systems or Workload Management Systems from 1983 to 2010 and applicable for inpatient medical/surgical settings were reviewed. Generally, we found that many of the criticisms of earlier PCSs are still evident: (1) difficulties with measuring workload remain an overarching theme throughout the literature; (2) definitions and descriptions of nursing work continue to be deemed inadequate; (3) there is insufficient evidence of reliability and validity testing of PCSs; and (4) there is still a need to identify nursing sensitive performance indicators and outcomes. We identified characteristics of promising PCSs, but concluded that no consensus exists about PCSs. We suggest that any approach to predicting staffing should seek to be parsimonious, minimize additional workload, be based on expert nurse judgment, be a true reflection of nursing work, and include indicators that measure patient complexity, required nursing care, available resources, and relevant organizational attributes. PMID- 21639032 TI - An integrative review of nursing workforce studies. AB - The purpose of this chapter is to present an analysis of selected published nursing workforce studies published between the years of 2005 and 2010. Thirteen nursing workforce studies were reviewed and analyzed using a modification of the method suggested by Ganong (1987). Nursing workforce studies were selected based on the following criteria: (1) the date of publication was between the years of 2005 and 2010; (2) the primary focus was on nurses working in practice; or, as students or faculty in nursing educational programs. When reviewed, the 13 studies (1) lacked uniform measures among databases; (2) lacked longitudinal studies that followed the respondent over time from the beginning of their career to retirement; (3) had response rates that contributed to small sample sizes or sampling frame that did not take into consideration all characteristics of interest; (4) lacked attention to an interdisciplinary mix of providers; and (5) implied the need for future study on intergenerational characteristics due to shifting demographics in the profession and nursing workforce. PMID- 21639033 TI - Carbon, nitrogen, and phosphorus fluxes in household ecosystems in the Minneapolis-Saint Paul, Minnesota, urban region. AB - Rapid worldwide urbanization calls for a better understanding of the biogeochemical cycling of those macroelements that have large environmental impacts in cities. This study, part of the Twin Cities Household Ecosystem Project, quantified fluxes of carbon (C), nitrogen (N), and phosphorus (P) at the scale of individual households in the Minneapolis-Saint Paul metropolitan area in Minnesota, USA. We estimated input and output fluxes associated with several components of household activities including air and motor vehicle travel, food consumption, home energy use, landscape, pets, and paper and plastic use for 360 owner-occupied, stand-alone households. A few component fluxes dominated total input fluxes of elements. For instance, air and motor vehicle transportation, together with home energy use, accounted for 85% of total C consumption and emissions. All total and component fluxes were skewed to varying degrees, suggesting that policies targeting disproportionately high fluxes could be an effective and efficient way to reduce pollution. For example, 20% of households contributed 75% of air travel emissions and 40% of motor vehicle emissions. Home energy use was more nearly normally distributed. Nitrogen fluxes were dominated by human diet and lawn fertilizer applications, which together accounted for 65% of total household N inputs. The majority of P inputs were associated with human diet, use of detergents, and pet food. A large portion of the variation among household fluxes of C, N, and P was related to a few biophysical variables. A better understanding of the biophysical, demographic, and behavioral drivers of household activities that contribute to C, N, and P fluxes is pivotal for developing accurate urban biogeochemical models and for informing policies aimed at reducing sources of pollution in urban ecosystems. PMID- 21639034 TI - Ecosystem response to nutrient enrichment across an urban airshed in the Sonoran Desert. AB - Rates of nitrogen (N) deposition have increased in arid and semiarid ecosystems, but few studies have examined the impacts of long-term N enrichment on ecological processes in deserts. We conducted a multiyear, nutrient-addition study within 15 Sonoran Desert sites across the rapidly growing metropolitan area of Phoenix, Arizona (USA). We hypothesized that desert plants and soils would be sensitive to N enrichment, but that these effects would vary among functional groups that differ in terms of physiological responsiveness, proximity to surface N sources, and magnitude of carbon (C) or water limitation. Inorganic N additions augmented net potential nitrification in soils, moreso than net potential N mineralization, highlighting the important role of nitrifying microorganisms in the nitrate economy of drylands. Winter annual plants were also responsive to nutrient additions, exhibiting a climate-driven cascade of resource limitation, from little to no production in seasons of low rainfall (winter 2006 and 2007), to moderate N limitation with average precipitation (winter 2009), to limitation by both N and P in a season of above-normal rainfall (winter 2008). Herbaceous production is a potentially important mechanism of N retention in arid ecosystems, capable of immobilizing an amount equal to or greater than that deposited annually to soils in this urban airshed. However, interannual variability in precipitation and abiotic processes that limit the incorporation of detrital organic matter into soil pools may limit this role over the long term. In contrast, despite large experimental additions of N and P over four years, growth of Larrea tridentata, the dominant perennial plant of the Sonoran Desert, was unresponsive to nutrient enrichment, even during wet years. Finally, there did not appear to be strong ecological interactions between nutrient addition and location relative to the city, despite the nearby activity of nearly four million people, perhaps due to loss or transfer pathways that limit long term N enrichment of ecosystems by the urban atmosphere. PMID- 21639035 TI - Transpiration of urban forests in the Los Angeles metropolitan area. AB - Despite its importance for urban planning, landscape management, and water management, there are very few in situ estimates of urban-forest transpiration. Because urban forests contain an unusual and diverse mix of species from many regions worldwide, we hypothesized that species composition would be a more important driver of spatial variability in urban-forest transpiration than meteorological variables in the Los Angeles (California, USA) region. We used constant-heat sap-flow sensors to monitor urban tree water use for 15 species at six locations throughout the Los Angeles metropolitan area. For many of these species no previous data on sap flux, water use, or water relations were available in the literature. To scale sap-flux measurements to whole trees we conducted a literature survey of radial trends in sap flux across multiple species and found consistent relationships for angiosperms vs. gymnosperms. We applied this relationship to our measurements and estimated whole-tree and plot level transpiration at our sites. The results supported very large species differences in transpiration, with estimates ranging from 3.2 +/- 2.3 kg x tree( 1) x d(-1) in unirrigated Pinus canariensis (Canary Island pine) to 176.9 +/- 75.2 kg x tree(-1) x d(-1) in Platanus hybrida (London planetree) in the month of August. Other species with high daily transpiration rates included Ficus microcarpa (laurel fig), Gleditsia triacanthos (honeylocust), and Platanus racemosa (California sycamore). Despite irrigation and relatively large tree size, Brachychiton populneas (kurrajong), B. discolor (lacebark), Sequoia sempervirens (redwood), and Eucalyptus grandis (grand Eucalyptus) showed relatively low rates of transpiration, with values < 45 kg x tree(-1) x d(-1). When scaled to the plot level, transpiration rates were as high as 2 mm/d for sites that contained both species with high transpiration rates and high densities of planted trees. Because plot-level transpiration is highly dependent on tree density, we modeled transpiration as a function of both species and density to evaluate a likely range of values in irrigated urban forests. The results show that urban forests in irrigated, semi-arid regions can constitute a significant use of water, but water use can be mitigated by appropriate selection of site, management method, and species. PMID- 21639036 TI - Water subsidies from mountains to deserts: their role in sustaining groundwater fed oases in a sandy landscape. AB - In arid regions throughout the world, shallow phreatic aquifers feed natural oases of much higher productivity than would be expected solely from local rainfall. In South America, the presence of well-developed Prosopis flexuosa woodlands in the Monte Desert region east of the Andes has puzzled scientists for decades. Today these woodlands provide crucial subsistence to local populations, including descendants of the indigenous Huarpes. We explore the vulnerability and importance of phreatic groundwater for the productivity of the region, comparing the contributions of local rainfall to that of remote mountain recharge that is increasingly being diverted for irrigated agriculture before it reaches the desert. We combined deep soil coring, plant measurements, direct water-table observations, and stable-isotopic analyses (2H and 18O) of meteoric, surface, and ground waters at three study sites across the region, comparing woodland stands, bare dunes, and surrounding shrublands. The isotopic composition of phreatic groundwaters (delta2H: -137 per thousand +/- 5 per thousand) closely matched the signature of water brought to the region by the Mendoza River (-137 per thousand +/- 6 per thousand), suggestin that mountain-river infiltration rather than in situ rainfall deep drainage (-39 per thousand +/- 19 per thousand) was the dominant mechanism of recharge. Similarly, chloride mass balances determined from deep soil profiles (> 6 m) suggested very low recharge rates. Vegetation in woodland ecosystems, where significant groundwater discharge losses, likely >100 mm/yr occurred, relied on regionally derived groundwater located from 6.5 to 9.5 m underground. At these locations, daily water-table fluctuations of 10 mm, and stable-isotopic measurements of plant water, indicated groundwater uptake rates of 200-300 mm/yr. Regional scaling suggests that groundwater evapotranspiration reaches 18-42 mm/yr across the landscape, accounting for 7 17% of the Mendoza River flow regionally. Our study highlights the reliance of ecosystem productivity in natural oases on Andean snowmelt, which is increasingly being diverted to one of the largest irrigated regions of the continent. Understanding the ecohydrological coupling of mountain and desert ecosystems here and elsewhere should help managers balance production agriculture and conservation of unique woodland ecosystems and the rural communities that rely on them. PMID- 21639037 TI - Litter quality impacts short- but not long-term soil carbon dynamics in soil aggregate fractions. AB - Complex molecules are presumed to be preferentially stabilized as soil organic carbon (SOC) based on the generally accepted concept that the chemical composition of litter is a major factor in its rate of decomposition. Hence, a direct link between litter quality and SOC quantity has been assumed, accepted, and ultimately incorporated in SOC models. Here, however, we present data from an incubation and field experiment that refutes the influence of litter quality on the quantity of stabilized SOC. Three different qualities of litter (Tithonia diversifolia, Calliandra calothyrsus, and Zea mays stover; 4 Mg C x ha(-1) yr( 1)) with and without the addition of mineral N fertilizer (0 or 120 kg N x ha( 1)season(-1) were added to a red clay Humic Nitisol in a 3-yr field trial and a 1.5-yr incubation experiment. The litters differed in their concentrations of N, lignin, and polyphenols with the ratio of (lignin + polyphenols): N ranging from 3.5 to 9.8 for the field trial and from 2.3 to 4.0 for the incubation experiment in the order of T. diversifolia < C. calothyrsus < or = Z. mays. Litter quality did not affect the amount of SOC stabilized after three annual additions in the field trial. Even within the most sensitive soil aggregate fractions, SOC contents and C:N ratios did not differ with litter quality, indicating that litter quality did not influence the mechanisms by which SOC was stabilized. While increasing litter quality displayed faster decomposition and incorporation of C into soil aggregates after 0.25 yr in the incubation study, all litters resulted in equivalent amounts of C stabilized in the soil after 1.5 yr, further corroborating the results of the field trial. The addition of N fertilizer did not affect SOC stabilization in either the field or the incubation trial. Thus, we conclude that, while litter quality controls shorter-term dynamics of C decomposition and accumulation in the soil, longer-term SOC patterns cannot be predicted based on initial litter quality effects. Hence, the formation and stabilization of SOC is more controlled by the quantity of litter input and its interaction with the soil matrix than by litter quality. PMID- 21639038 TI - Regional and decadal patterns of native and exotic plant coexistence in California grasslands. AB - Coexistence through a variety of mechanisms is possible for species with differential responses to environmental conditions. Understanding the role of environmental heterogeneity in mediating coexistence of species of different provenance (i.e., native vs. exotic) has important implications for theory and management. We used two California grassland data sets, one spanning seven years at three reserves along a 500-km latitudinal gradient and one spanning 47 years at 11 sites within a single 1000-ha reserve, to determine how environmental heterogeneity in space and time contributes to variability in provenance group abundance and diversity, and whether native and exotic species respond similarly to spatial and temporal variability. We found that temporal environmental heterogeneity is the primary determinant of provenance group abundance, while spatial and temporal environmental heterogeneity both contribute to community diversity. Spatial and temporal heterogeneity must therefore be considered simultaneously when examining community dynamics and species coexistence. Provenance was a poor general predictor of species response; native and exotic species exhibit similar spatiotemporal patterns in some cases but not others. Plant persistence may depend more upon the abiotic environment than competition from the other provenance group as native and exotic diversity were generally positively correlated. Furthermore, mesoscale (10(2)-10(3) m) spatial heterogeneity may be a greater mediator of provenance group coexistence than temporal heterogeneity or spatial heterogeneity at other scales. PMID- 21639039 TI - Restoring diversity after cattail expansion: disturbance, resilience, and seasonality in a tropical dry wetland. AB - As the human footprint expands, ecologists and resource managers are increasingly challenged to explain and manage abrupt ecosystem transformations (i.e., regime shifts). In this study, we investigated the role of a mechanical disturbance that has been used to restore and maintain local wetland diversity after a monotypic regime shift in northwestern Costa Rica [specifically, an abrupt landscape-scale cattail (Typha) expansion]. The study was conducted in Palo Verde Marsh (Palo Verde National Park; a RAMSAR Wetland of International Importance), a seasonally flooded freshwater wetland that has historically provided habitat for large populations of wading birds and waterfowl. A cattail (T. domingensis) expansion in the 1980s greatly altered the plant community and reduced avian habitat. Since then, Typha has been managed using a form of mechanical disturbance called fangueo (a Spanish word, pronounced "fahn-gay-yo" in English). We applied a Typha removal treatment at three levels (control, fangueo, and fangueo with fencing to exclude cattle grazing). Fangueo resulted in a large reduction in Typha dominance (i.e., decreased aboveground biomass, ramet density, and ramet height) and an increase in habitat heterogeneity. As in many ecosystems that have been defined by multiple and frequent disturbances, a large portion of the plant community regenerated after disturbance (via propagule banking) and fangueo resulted in a more diverse plant community that was strongly dictated by seasonal processes (i.e., distinct wet- and dry-season assemblages). Importantly, the mechanical disturbance had no apparent short-term impact on any of the soil properties we measured (including bulk density). Interestingly, low soil and foliar N:P values indicate that Palo Verde Marsh and other wetlands in the region may be nitrogen limited. Our results quantify how, in a cultural landscape where the historical disturbance regime has been altered and diversity has declined, a mechanical disturbance in combination with seasonal drought and flooding has been used to locally restrict a clonal monodominant plant expansion, create habitat heterogeneity, and maintain plant diversity. PMID- 21639040 TI - Lower resistance and higher tolerance of invasive host plants: biocontrol agents reach high densities but exert weak control. AB - Invasive plants often have novel biotic interactions in their introduced ranges. Their defense to herbivory may differ from their native counterparts, potentially influencing the effectiveness of biological control. If invasive plants have decreased resistance but increased tolerance to enemies, insect herbivores may rapidly build up their populations but exert weak control. Moreover, resource availability to plants may affect the efficacy of biological control agents. We tested these predictions using Chinese tallow tree (Triadica sebifera) and two specialist herbivores (Heterapoderopsis bicallosicollis and Gadirtha inexacta) that are candidates for biological control. We performed a pair of field common garden experiments in China in which Triadica seedlings from the native or introduced range were grown in low or high light conditions and subjected to different levels of herbivory by each herbivore in a factorial design. We found that Heterapoderopsis achieved greater densities on tallow trees from the introduced range or when trees were grown in high light conditions. When Gadirtha was raised in the lab on tallow tree foliage we found that it performed better (larger pupal size) when fed foliage from introduced populations. However, introduced populations generally had greater herbivore tolerance such that the impact of each agent on plant performance was lower than on native populations despite higher herbivore loads. Tallow trees grew more slowly and achieved smaller sizes in lower light levels, but the impact of biological control agents was comparable to that found for higher light levels. Plants from introduced populations grew larger than those from native populations in all conditions. Our results suggest that reduced resistance and increased tolerance to herbivory in introduced populations may impede success of biological control programs. Biological control practitioners should include plants from the introduced range in the prerelease evaluation, which will help predict insect impact on target weeds. PMID- 21639041 TI - Landscape connectivity and seed dispersal characteristics inform the best management strategy for exotic plants. AB - Exotic plant invasions have triggered environmental and economic problems throughout the world. Our ability to manage these invasions is hindered by the difficulty of predicting spread in fragmented landscapes. Because the spatial pattern of invasions depends on the dispersal characteristics of the invasive species and the configuration of suitable habitat within the landscape, a universal management strategy is unlikely to succeed for any particular species. We suggest that the most effective management strategy may be an adaptive one that shifts from local control to landscape management depending on the specific invader and landscape. In particular, we addressed the question of where management activities should be focused to minimize spread of the invading species. By simulating an invasion across a real landscape (Antietam National Battlefield in Maryland, USA), we examined the importance of patch size and connectivity to management success. We found that the best management strategy depended on the dispersal characteristics of the exotic species. Species with a high probability of random long-distance dispersal were best managed by focusing on the largest patches, while species with a lower probability of random long distance dispersal were best managed by considering landscape configuration and connectivity of the patches. Connectivity metrics from network analysis were useful for identifying the most effective places to focus management efforts. These results provide insight into invasion patterns of various species and suggest a general rule for managers in National Parks and other places where invasive species are a concern. PMID- 21639042 TI - Historical carbon emissions and uptake from the agricultural frontier of the Brazilian Amazon. AB - Tropical ecosystems play a large and complex role in the global carbon cycle. Clearing of natural ecosystems for agriculture leads to large pulses of CO2 to the atmosphere from terrestrial biomass. Concurrently, the remaining intact ecosystems, especially tropical forests, may be sequestering a large amount of carbon from the atmosphere in response to global environmental changes including climate changes and an increase in atmospheric CO2. Here we use an approach that integrates census-based historical land use reconstructions, remote-sensing-based contemporary land use change analyses, and simulation modeling of terrestrial biogeochemistry to estimate the net carbon balance over the period 1901-2006 for the state of Mato Grosso, Brazil, which is one of the most rapidly changing agricultural frontiers in the world. By the end of this period, we estimate that of the state's 925 225 km2, 221 092 km2 have been converted to pastures and 89 533 km2 have been converted to croplands, with forest-to-pasture conversions being the dominant land use trajectory but with recent transitions to croplands increasing rapidly in the last decade. These conversions have led to a cumulative release of 4.8 Pg C to the atmosphere, with 80% from forest clearing and 20% from the clearing of cerrado. Over the same period, we estimate that the residual undisturbed ecosystems accumulated 0.3 Pg C in response to CO2 fertilization. Therefore, the net emissions of carbon from Mato Grosso over this period were 4.5 Pg C. Net carbon emissions from Mato Grosso since 2000 averaged 146 Tg C/yr, on the order of Brazil's fossil fuel emissions during this period. These emissions were associated with the expansion of croplands to grow soybeans. While alternative management regimes in croplands, including tillage, fertilization, and cropping patterns promote carbon storage in ecosystems, they remain a small portion of the net carbon balance for the region. This detailed accounting of a region's carbon balance is the type of foundation analysis needed by the new United Nations Collaborative Programmme for Reducing Emissions from Deforestation and Forest Degradation (REDD). PMID- 21639043 TI - Fire regime in a Mexican forest under indigenous resource management. AB - The Raramuri (Tarahumara) people live in the mountains and canyons of the Sierra Madre Occidental of Chihuahua, Mexico. They base their subsistence on multiple use strategies of their natural resources, including agriculture, pastoralism, and harvesting of native plants and wildlife. Pino Gordo is a Raramuri settlement in a remote location where the forest has not been commercially logged. We reconstructed the forest fire regime from fire-scarred trees, measured the structure of the never-logged forest, and interviewed community members about fire use. Fire occurrence was consistent throughout the 19th and 20th centuries up to our fire scar collection in 2004. This is the least interrupted surface fire regime reported to date in North America. Studies from other relict sites such as nature reserves in Mexico or the USA have all shown some recent alterations associated with industrialized society. At Pino Gordo, fires recurred frequently at the three study sites, with a composite mean fire interval of 1.9 years (all fires) to 7.6 years (fires scarring 25% or more of samples). Per sample fire intervals averaged 10-14 years at the three sites. Approximately two thirds of fires burned in the season of cambial dormancy, probably during the pre monsoonal drought. Forests were dominated by pines and contained many large living trees and snags, in contrast to two nearby similar forests that have been logged. Community residents reported using fire for many purposes, consistent with previous literature on fire use by indigenous people. Pino Gordo is a valuable example of a continuing frequent-fire regime in a never-harvested forest. The Raramuri people have actively conserved this forest through their traditional livelihood and management techniques, as opposed to logging the forest, and have also facilitated the fire regime by burning. The data contribute to a better understanding of the interactions of humans who live in pine forests and the fire regimes of these ecosystems, a topic that has been controversial and difficult to assess from historical or paleoecological evidence. PMID- 21639044 TI - Modeling lodgepole pine radial growth relative to climate and genetics using universal growth-trend response functions. AB - Forests strongly affect Earth's carbon cycles, making our ability to forecast forest-productivity changes associated with rising temperatures and changes in precipitation increasingly critical. In this study, we model the influence of climate on annual radial growth using lodgepole pine (Pinus contorta) trees grown for 34 years in a large provenance experiment in western Canada. We use a random coefficient modeling approach to build universal growth-trend response functions that simultaneously incorporate the impacts of different provenance and site climates on radial growth trends under present and future annual (growth-year), summer, and winter climate regimes. This approach provides new depth to traditional quantitative genetics population response functions by illustrating potential changes in population dominance over time, as well as indicating the age and size at which annual growth begins declining for any population growing in any location under any present or future climate scenario within reason, given the ages and climatic conditions sampled. Our models indicate that lodgepole pine radial-growth levels maximize between 3.9 degrees and 5.1 degrees C mean growth year temperature. This translates to productivity declining by the mid-21st century in southern and central British Columbia (BC), while increasing beyond the 2080s in northern BC and Yukon, as temperatures rise. Relative to summer climate indices, productivity is predicted to decline continuously through the 2080s in all locations, while relative to winter climate variables, the opposite trend occurs, with the growth increases caused by warmer winters potentially offsetting the summer losses. Trees from warmer provenances, i.e., from the center of the species range, perform best in nearly all of our present and future climate-scenario models. We recommend that similar models be used to analyze population growth trends relative to annual and intra-annual climate in other large-scale provenance trials worldwide. An open-access growth-trend data set encompassing numerous biomes, species, and provenances would contribute substantially to predicting forest productivity under future-climate scenarios. PMID- 21639045 TI - Scale-dependent controls on the area burned in the boreal forest of Canada, 1980 2005. AB - In the boreal forest of North America, as in any fire-prone biome, three environmental factors must coincide for a wildfire to occur: an ignition source, flammable vegetation, and weather that is conducive to fire. Despite recent advances, the relative importance of these factors remains the subject of some debate. The aim of this study was to develop models that identify the environmental controls on spatial patterns in area burned for the period 1980 2005 at several spatial scales in the Canadian boreal forest. Boosted regression tree models were built to relate high-resolution data for area burned to an array of explanatory variables describing ignitions, vegetation, and long-term patterns in fire-conducive weather (i.e., fire climate) at four spatial scales (10(2) km2, 10(3) km2, 10(4) km2, and 10(5) km2). We evaluated the relative contributions of these controls on area burned, as well as their functional relationships, across spatial scales. We also assessed geographic patterns of the influence of wildfire controls. The results indicated that extreme temperature during the fire season was a top control at all spatial scales, followed closely by a wind-driven index of ease of fire spread. However, the contributions of some variables differed substantially among the spatial scales, as did their relationship to area burned. In fact, for some key variables the polarity of relationships was inverted from the finest to the broadest spatial scale. It was difficult to unequivocally attribute values of relative importance to the variables chosen to represent ignitions, vegetation, and climate, as the interdependence of these factors precluded clear partitioning. Furthermore, the influence of a variable on patterns of area burned often changed enormously across the biome, which supports the idea that fire-environment relationships in the boreal forest are complex and nonstationary. PMID- 21639046 TI - Managing the forest for more than the trees: effects of experimental timber harvest on forest Lepidoptera. AB - Studies of the effects of timber harvest on forest insect communities have rarely considered how disturbance from a range of harvest levels interacts with temporal variation in species diversity to affect community resistance to change. Here I report the results of a landscape-scale, before-and-after, treatment-control experiment designed to test how communities of forest Lepidoptera experience (1) changes in species richness and composition and (2) shifts in species dominance one year after logging. I sampled Lepidoptera from 20 forest stands allocated to three harvest treatments (control, even-aged shelterwood or clearcuts, and uneven aged group selection cuts) within three watersheds at Morgan-Monroe State Forest, Indiana, USA. Moths were sampled from all forest stands one year prior to harvest in 2007 and immediately post-harvest in 2009. Species composition was most significantly affected by temporal variation between years, although uneven-aged management also caused significant changes in lepidopteran community structure. Furthermore, species richness of Lepidoptera was higher in 2007 compared to 2009 across all watersheds and forest stands. The decrease in species richness between years, however, was much larger in even-aged and uneven-aged management units compared to the control. Furthermore, matrix stands within the even-aged management unit demonstrated the highest resistance to species loss within any management unit. Species dominance was highly resistant to effects of timber harvest, with pre- and post-harvest values for Simpson diversity nearly invariant. Counter to prediction, however, the suite of dominant taxa differed dramatically among the three management units post-harvest. My results suggest that temporal variation may have strong interactions with timber harvest, precipitating loss of nearly 50% species richness from managed stands regardless of harvest level. Even-aged management, however, appeared to leave the smallest "footprint" on moth communities. Timber harvest also created greater heterogeneity in species dominance among management units, suggesting that forest watersheds may diverge in post-harvest recovery transients relative to unlogged watersheds in Morgan-Monroe State Forest. Because moth communities appear to be highly sensitive to group-selection cuts, shelterwood cuts, and clearcuts, it will be important to retain unlogged concessions within management units to resist landscape-level loss of Lepidoptera. PMID- 21639047 TI - Passive sampling effects and landscape location alter associations between species traits and response to fragmentation. AB - As tropical reserves become smaller and more isolated, the ability of species to utilize fragmented landscapes will be a key determinant of species survival. Although several ecological and life history traits commonly are associated with vulnerability to fragmentation, the combination of traits that are most highly influential and the effectiveness of those traits in predicting vulnerability across distinct landscapes, remains poorly understood. We studied use of forest fragments by 25 mid- and large-sized neotropical mammals in Guatemala to determine how seven species traits influence vulnerability to fragmentation. We measured vulnerability in two ways: one measure that did not remove passive sampling effects (proportion of fragments occupied), and one that did (difference in occupancy rates within continuous and fragmented sites). We also examined the influence of species traits on patch occupancy rates of the same set of mammals on two landscapes in Mexico. When not accounting for passive sampling effects, body size, home range size, and vulnerability to hunting influenced how species responded to fragmentation. However, after controlling for passive sampling effects, only vulnerability to hunting strongly influenced sensitivity to fragmentation. Species that were heavily hunted were much less common in forest patches than in continuous forest sites of the same sampling size. The cross landscape comparison revealed both similarities and differences in the species traits that influenced patch occupancy patterns on each landscape. Given the ubiquity of hunting in tropical environments, our findings indicate that management efforts in fragmented landscapes that do not account for hunting pressure may be ineffective in conserving heavily hunted tropical species. Our study also indicates that species traits may be useful in predicting relative patch occupancy rates and/or vulnerability to fragmentation across distinct landscapes, but that caution must be used as certain traits can become more or less influential on different landscapes, even when considering the same set of species. PMID- 21639048 TI - Predicting potential European bison habitat across its former range. AB - Habitat loss threatens large mammals worldwide, and their survival will depend on habitat in human-dominated landscapes. Conservation planners thus face the challenge to identify areas of least conflict with land use, yet broadscale species distribution models rarely incorporate real landscape patterns nor do they identify potential conservation conflicts. An excellent example of such conservation challenges is provided by European bison (Bison bonasus). Almost extinct by the early 20th century, bison can only survive in the wild if large metapopulations are established, but it is unclear where new herds can be reintroduced. Using European bison as an example we conducted a continental-scale habitat assessment based on real landscape patterns. Our specific aims here were to (1) map European bison habitat throughout the species' former range, (2) examine whether broadscale habitat suitability factors differ from previously reported fine-scale factors, and (3) assess where suitable habitat occurs in areas with low potential for conflict with land use. We assessed habitat suitability using herd range maps for all 36 free-ranging European bison herds as habitat use data. Habitat suitability maps were compared with maps of land cover, livestock density, agricultural constraints, and protected areas to assess potential conservation conflicts. Our models had high goodness of fit (AUC = 0.941), and we found abundant potential bison habitat. European bison prefer mosaic-type landscapes, with a preference for broad-leaved and mixed forests. European bison metapopulations do not appear to be limited by habitat availability. However, most potential habitat occurred outside protected areas and has substantial potential for conservation conflicts. The most promising areas for establishing large bison metapopulations all occur in Eastern Europe (i.e., the Carpathians, the Belarus-Ukraine borderlands, and several regions in European Russia). The future of European bison and that of other large mammals in the wild thus clearly lies in Eastern Europe, because habitat there is most abundant and least fragmented, and because the potential for conflict with land use is lower. More generally we suggest that broadscale habitat assessments that incorporate land use can be powerful tools for conservation planning and will be key if large herbivore and carnivore conservation is to succeed in a human dominated world. PMID- 21639049 TI - Allocating conservation resources between areas where persistence of a species is uncertain. AB - Research on the allocation of resources to manage threatened species typically assumes that the state of the system is completely observable; for example whether a species is present or not. The majority of this research has converged on modeling problems as Markov decision processes (MDP), which give an optimal strategy driven by the current state of the system being managed. However, the presence of threatened species in an area can be uncertain. Typically, resource allocation among multiple conservation areas has been based on the biggest expected benefit (return on investment) but fails to incorporate the risk of imperfect detection. We provide the first decision-making framework for confronting the trade-off between information and return on investment, and we illustrate the approach for populations of the Sumatran tiger (Panthera tigris sumatrae) in Kerinci Seblat National Park. The problem is posed as a partially observable Markov decision process (POMDP), which extends MDP to incorporate incomplete detection and allows decisions based on our confidence in particular states. POMDP has previously been used for making optimal management decisions for a single population of a threatened species. We extend this work by investigating two populations, enabling us to explore the importance of variation in expected return on investment between populations on how we should act. We compare the performance of optimal strategies derived assuming complete (MDP) and incomplete (POMDP) observability. We find that uncertainty about the presence of a species affects how we should act. Further, we show that assuming full knowledge of a species presence will deliver poorer strategic outcomes than if uncertainty about a species status is explicitly considered. MDP solutions perform up to 90% worse than the POMDP for highly cryptic species, and they only converge in performance when we are certain of observing the species during management: an unlikely scenario for many threatened species. This study illustrates an approach to allocating limited resources to threatened species where the conservation status of the species in different areas is uncertain. The results highlight the importance of including partial observability in future models of optimal species management when the species of concern is cryptic in nature. PMID- 21639050 TI - Polar bear population status in the northern Beaufort Sea, Canada, 1971-2006. AB - Polar bears (Ursus maritimus) of the northern Beaufort Sea (NB) population occur on the perimeter of the polar basin adjacent to the northwestern islands of the Canadian Arctic Archipelago. Sea ice converges on the islands through most of the year. We used open-population capture-recapture models to estimate population size and vital rates of polar bears between 1971 and 2006 to: (1) assess relationships between survival, sex and age, and time period; (2) evaluate the long-term importance of sea ice quality and availability in relation to climate warming; and (3) note future management and conservation concerns. The highest ranking models suggested that survival of polar bears varied by age class and with changes in the sea ice habitat. Model-averaged estimates of survival (which include harvest mortality) for senescent adults ranged from 0.37 to 0.62, from 0.22 to 0.68 for cubs of the year (COY) and yearlings, and from 0.77 to 0.92 for 2-4 year-olds and adults. Horvtiz-Thompson (HT) estimates of population size were not significantly different among the decades of our study. The population size estimated for the 2000s was 980 +/- 155 (mean and 95% CI). These estimates apply primarily to that segment of the NB population residing west and south of Banks Island. The NB polar bear population appears to have been stable or possibly increasing slightly during the period of our study. This suggests that ice conditions have remained suitable and similar for feeding in summer and fall during most years and that the traditional and legal Inuvialuit harvest has not exceeded sustainable levels. However, the amount of ice remaining in the study area at the end of summer, and the proportion that continues to lie over the biologically productive continental shelf (< 300 m water depth) has declined over the 35-year period of this study. If the climate continues to warm as predicted, we predict that the polar bear population in the northern Beaufort Sea will eventually decline. Management and conservation practices for polar bears in relation to both aboriginal harvesting and offshore industrial activity will need to adapt. PMID- 21639051 TI - How many genetic markers to tag an individual? An empirical assessment of false matching rates among close relatives. AB - Genetic identification of individuals is now commonplace, enabling the application of tagging methods to elusive species or species that cannot be tagged by traditional methods. A key aspect is determining the number of loci required to ensure that different individuals have non-matching multi-locus genotypes. Closely related individuals are of particular concern because of elevated matching probabilities caused by their recent co-ancestry. This issue may be addressed by increasing the number of loci to a level where full siblings (the relatedness category with the highest matching probability) are expected to have non-matching multi-locus genotypes. However, increasing the number of loci to meet this "full-sib criterion" greatly increases the laboratory effort, which in turn may increase the genotyping error rate resulting in an upward-biased mark recapture estimate of abundance as recaptures are missed due to genotyping errors. We assessed the contribution of false matches from close relatives among 425 maternally related humpback whales, each genotyped at 20 microsatellite loci. We observed a very low (0.5-4%) contribution to falsely matching samples from pairs of first-order relatives (i.e., parent and offspring or full siblings). The main contribution to falsely matching individuals from close relatives originated from second-order relatives (e.g., half siblings), which was estimated at 9%. In our study, the total number of observed matches agreed well with expectations based upon the matching probability estimated for unrelated individuals, suggesting that the full-sib criterion is overly conservative, and would have required a 280% relative increase in effort. We suggest that, under most circumstances, the overall contribution to falsely matching samples from close relatives is likely to be low, and hence applying the full-sib criterion is unnecessary. In those cases where close relatives may present a significant issue, such as unrepresentative sampling, we propose three different genotyping strategies requiring only a modest increase in effort, which will greatly reduce the number of false matches due to the presence of related individuals. PMID- 21639052 TI - Historical and contemporary trophic niche partitioning among Laurentian Great Lakes coregonines. AB - Anthropogenic activities have significantly altered freshwater fish communities. Extirpations of deepwater coregonines (Coregonus spp.), a diverse group of fish species, have left vast areas of the Laurentian Great Lakes devoid of a deepwater fish community. Currently, fisheries managers are considering restoring populations by reintroducing deepwater coregonines from Lake Superior and Lake Nipigon. However, little is known about the historical ecology of deepwater coregonines, and species characterization has proved difficult. We used stable isotope analysis of museum-preserved and contemporary specimens to investigate if (1) coregonine species historically occupied distinct niches and (2) the pattern of trophic niche partitioning has changed over the last century. Across all lakes, individual species occupied distinct trophic niches, confirming that these species were ecologically distinct. Understanding trophic niche partitioning helps resolve uncertainty about distinctness of species within and across lakes and may provide a better ecological basis for rehabilitation of Great Lakes food webs and ecosystems. PMID- 21639053 TI - Fish farms, parasites, and predators: implications for salmon population dynamics. AB - For some salmon populations, the individual and population effects of sea lice (Lepeophtheirus salmonis) transmission from sea cage salmon farms is probably mediated by predation, which is a primary natural source of mortality of juvenile salmon. We examined how sea lice infestation affects predation risk and mortality of juvenile pink (Oncorhynchus gorbuscha) and chum (O. keta) salmon, and developed a mathematical model to assess the implications for population dynamics and conservation. A risk-taking experiment indicated that infected juvenile pink salmon accept a higher predation risk in order to obtain foraging opportunities. In a schooling experiment with juvenile chum salmon, infected individuals had increased nearest-neighbor distances and occupied peripheral positions in the school. Prey selection experiments with cutthroat trout (O. clarkii) predators indicated that infection reduces the ability of juvenile pink salmon to evade a predatory strike. Group predation experiments with coho salmon (O. kisutch) feeding on juvenile pink or chum salmon indicated that predators selectively consume infected prey. The experimental results indicate that lice may increase the rate of prey capture but not the handling time of a predator. Based on this result, we developed a mathematical model of sea lice and salmon population dynamics in which parasitism affects the attack rate in a type II functional response. Analysis of the model indicates that: (1) the estimated mortality of wild juvenile salmon due to sea lice infestation is probably higher than previously thought; (2) predation can cause a simultaneous decline in sea louse abundance on wild fish and salmon productivity that could mislead managers and regulators; and (3) compensatory mortality occurs in the saturation region of the type II functional response where prey are abundant because predators increase mortality of parasites but not overall predation rates. These findings indicate that predation is an important component of salmon-louse dynamics and has implications for estimating mortality, reducing infection, and developing conservation policy. PMID- 21639054 TI - Modeling the impacts of the European green crab on commercial shellfisheries. AB - Coastal resource managers are often tasked with managing coastal ecosystems that are stressed by overexploitation, climate change, contaminants, and habitat loss, as well as biological invasions. Therefore, managers increasingly need better economic data to help them prioritize their management strategies and distribute their increasingly limited resources to those strategies. Despite frequent pronouncements about the substantial ecological and economic impacts of invasive species, there have been few if any rigorous analyses of the economic impacts of invasive species in coastal systems. Here we present a bioeconomic analysis of the impacts of the European green crab, Carcinus maenas, on commercial shellfisheries along the West Coast of the United States. Green crabs are among the most comprehensively studied and widely distributed invasive species in coastal systems, with established populations on every continent except Antarctica. Their impacts on commercial bivalve fisheries have been alleged or substantiated to varying degrees, but no formal analysis of the economic impacts of the green crab has been conducted. We assess economic impacts using a combination of ecological and economic models. The ecological models incorporate green crab dispersal and description of estuarine habitat and the relationship between green crab abundance and predation on prey populations. The economic analysis focuses on the green crab impacts on commercial shellfisheries, including both historical and present impacts of green crabs on several important shellfisheries, including soft-shell clams, blue mussels, scallops, hard-shell clams, and Manila clams. We conclude that the past and present economic impacts on the West Coast shellfisheries are minor, although losses could increase significantly if densities increase or with northward range expansion into Alaska. PMID- 21639055 TI - Changes in habitat heterogeneity alter marine sessile benthic communities. AB - Habitat heterogeneity is considered an important mechanism influencing diversity patterns in spatially structured habitats. However, spatial heterogeneity is not static and it can change along temporal scales. These changes, whether gradual or rapid, have the potential of forcing species extinctions or facilitating the introduction of nonnative species. Here, we present modeling results that show how changes in spatial heterogeneity over several generations can produce strong changes in benthic species composition residing in eastern Long Island Sound, USA. For many benthic species, hard substrate is a limiting resource which can vary in availability among different coastal areas. We modeled gradual changes from a heterogeneous landscape (mimicking patches of natural hard and soft substrate) to a homogenous one (analogous to a fully developed coast with hard, manmade substrate) and followed the abundance and distribution patterns of species possessing four different life histories. We also modeled changes from homogeneous to heterogeneous landscapes. We found that as regions become more homogeneous, species extinctions become more frequent and poor dispersers dominate locally. In contrast, as habitats become more heterogeneous, species distributing across localities leads to regional species coexistence and fewer extinctions. These results suggest that focusing on changing habitat heterogeneity can be a useful management strategy to prevent poor dispersing species, such as invasive ascidians, from driving communities to monocultures. PMID- 21639056 TI - Anthropogenic resource subsidies decouple predator-prey relationships. AB - The extent to which resource subsidies affect food web dynamics is poorly understood in anthropogenic landscapes. To better understand how species interactions are influenced by subsidies, we studied breeding birds and nest predators along a rural-to-urban landscape gradient that varied in subsidies provided to generalist predators. We hypothesized that resource subsidies in urban landscapes would decouple predator-prey relationships, as predators switch from natural to anthropogenic foods. From 2004 to 2009, we surveyed nest predators and monitored 2942 nests of five songbird species breeding in 19 mature forest stands in Ohio, USA. Eighteen species were video-recorded depredating nests. Numbers of avian and mammalian nest predators were positively associated with the amount of urban development surrounding forests, with the exception of Brown-headed Cowbirds (Molothrus ater). Although nest survival strongly declined with detections of nest predators in rural landscapes, nest survival and predator numbers were unrelated in urban landscapes. Thus, the strength of interaction between breeding birds and nest predators diminished as landscapes surrounding forested parks became more urbanized. Our work suggests that decoupling of predator-prey relationships can arise when synanthropic predators are heavily subsidized by anthropogenic resources. In this way, human drivers can alter, and completely disarticulate, relationships among species that are well established in more natural systems. PMID- 21639057 TI - Fine-scale movement decisions of tropical forest birds in a fragmented landscape. AB - The persistence of forest-dependent species in fragmented landscapes is fundamentally linked to the movement of individuals among subpopulations. The paths taken by dispersing individuals can be considered a series of steps built from individual route choices. Despite the importance of these fine-scale movement decisions, it has proved difficult to collect such data that reveal how forest birds move in novel landscapes. We collected unprecedented route information about the movement of translocated forest birds from two species in the highly fragmented tropical dry forest of Costa Rica. In this pasture dominated landscape, forest remains in patches or riparian corridors, with lesser amounts of living fencerows and individual trees or "stepping stones." We used step selection functions to quantify how route choice was influenced by these habitat elements. We found that the amount of risk these birds were willing to take by crossing open habitat was context dependent. The forest-specialist Barred Antshrike (Thamnophilus doliatus) exhibited stronger selection for forested routes when moving in novel landscapes distant from its territory relative to locations closer to its territory. It also selected forested routes when its step originated in forest habitat. It preferred steps ending in stepping stones when the available routes had little forest cover, but avoided them when routes had greater forest cover. The forest-generalist Rufous-naped Wren (Campylorhynchus rufinucha) preferred steps that contained more pasture, but only when starting from non-forest habitats. Our results showed that forested corridors (i.e., riparian corridors) best facilitated the movement of a sensitive forest specialist through this fragmented landscape. They also suggested that stepping stones can be important in highly fragmented forests with little remaining forest cover. We expect that naturally dispersing birds and species with greater forest dependence would exhibit even stronger selection for forested routes than did the birds in our experiments. PMID- 21639058 TI - Spatially explicit modeling of conflict zones between wildlife and snow sports: prioritizing areas for winter refuges. AB - Outdoor winter recreation exerts an increasing pressure upon mountain ecosystems, with unpredictable, free-ranging activities (e.g., ski mountaineering, snowboarding, and snowshoeing) representing a major source of stress for wildlife. Mitigating anthropogenic disturbance requires the spatially explicit prediction of the interference between the activities of humans and wildlife. We applied spatial modeling to localize conflict zones between wintering Black Grouse (Tetrao tetrix), a declining species of Alpine timberline ecosystems, and two free-ranging winter sports (off-piste skiing [including snow-boarding] and snowshoeing). Track data (snow-sports and birds' traces) obtained from aerial photographs taken over a 585-km transect running along the timberline, implemented within a maximum entropy model, were used to predict the occurrence of snow sports and Black Grouse as a function of landscape characteristics. By modeling Black Grouse presence in the theoretical absence of free-ranging activities and ski infrastructure, we first estimated the amount of habitat reduction caused by these two factors. The models were then extrapolated to the altitudinal range occupied by Black Grouse, while the spatial extent and intensity of potential conflict were assessed by calculating the probability of human-wildlife co-occurrence. The two snow-sports showed different distribution patterns. Skiers' occurrence was mainly determined by ski-lift presence and a smooth terrain, while snowshoers' occurrence was linked to hiking or skiing routes and moderate slopes. Wintering Black Grouse avoided ski lifts and areas frequented by free-ranging snow sports. According to the models, Black Grouse have faced a substantial reduction of suitable wintering habitat along the timberline transect: 12% due to ski infrastructure and another 16% when adding free-ranging activities. Extrapolating the models over the whole study area results in an overall habitat loss due to ski infrastructure of 10%, while there was a > 10% probability of human-wildlife encounters on 67% of the remaining area of suitable wintering habitat. Only 23% of the wintering habitat was thus free of anthropogenic disturbance. By identifying zones of potential conflict, while rating its relative intensity, our model provides a powerful tool to delineate and prioritize areas where wildlife winter refuges and visitor steering measures should be implemented. PMID- 21639059 TI - Wetland features and landscape context predict the risk of wetland habitat loss. AB - Wetlands generally provide significant ecosystem services and function as important harbors of biodiversity. To ensure that these habitats are conserved, an efficient means of identifying wetlands at risk of conversion is needed, especially in the southern United States where the rate of wetland loss has been highest in recent decades. We used multivariate adaptive regression splines to develop a model to predict the risk of wetland habitat loss as a function of wetland features and landscape context. Fates of wetland habitats from 1992 to 1997 were obtained from the National Resources Inventory for the U.S. Forest Service's Southern Region, and land-cover data were obtained from the National Land Cover Data. We randomly selected 70% of our 40 617 observations to build the model (n = 28 432), and randomly divided the remaining 30% of the data into five Test data sets (n = 2437 each). The wetland and landscape variables that were important in the model, and their relative contributions to the model's predictive ability (100 = largest, 0 = smallest), were land-cover/ land-use of the surrounding landscape (100.0), size and proximity of development patches within 570 m (39.5), land ownership (39.1), road density within 570 m (37.5), percent woody and herbaceous wetland cover within 570 m (27.8), size and proximity of development patches within 5130 m (25.7), percent grasslands/herbaceous plants and pasture/hay cover within 5130 m (21.7), wetland type (21.2), and percent woody and herbaceous wetland cover within 1710 m (16.6). For the five Test data sets, Kappa statistics (0.40, 0.50, 0.52, 0.55, 0.56; P < 0.0001), area-under-the-receiver-operating-curve (AUC) statistics (0.78, 0.82, 0.83, 0.83, 0.84; P < 0.0001), and percent correct prediction of wetland habitat loss (69.1, 80.4, 81.7, 82.3, 83.1) indicated the model generally had substantial predictive ability across the South. Policy analysts and land-use planners can use the model and associated maps to prioritize at-risk wetlands for protection, evaluate wetland habitat connectivity, predict future conversion of wetland habitat based on projected land-use trends, and assess the effectiveness of wetland conservation programs. PMID- 21639060 TI - The distance that contaminated aquatic subsidies extend into lake riparian zones. AB - Consumption of emergent aquatic insects by terrestrial invertebrates is a poorly resolved, but potentially important, mechanism of contaminant flux across ecosystem borders leading to contaminant exposure in terrestrial invertivores. We characterized the spatial extent and magnitude of contaminant transfer from aquatic sediments to terrestrial invertebrate predators by examining riparian araneid spiders, terrestrial insects, and emergent aquatic insects for stable isotopes and polychlorinated biphenyls (PCBs, sum of 141 congeners) at Lake Hartwell, (Clemson, South Carolina, USA). PCB concentrations in aquatic insects were orders of magnitude higher than in terrestrial insects. Aquatic insect consumption by spiders (as indicated by delta13C and delta15N), PCB concentrations in spiders, and aquatic prey availability were greatest at the shoreline and declined inland, while terrestrial prey availability was invariant with distance. These patterns indicate PCB transfer to spiders through consumption of emergent aquatic insects extending to a distance of 5 m inland. Measurable, but much lower, PCBs were present in insect predators dominated by social wasps up to 30 m inland. These results illustrate the importance of emergent insects as vectors of contaminant transfer from lake sediments to riparian food webs, and that spiders are key predators in this process. PMID- 21639061 TI - Decoupled effects (positive to negative) of nutrient enrichment on ecosystem services. AB - Eutrophication is a widespread phenomenon that disrupts natural ecosystems around the globe. Despite the general recognition that ecosystems provide many services and benefits to humans, little effort has been made to address how increasing anthropogenic eutrophication affects those services. We conducted a field experiment to determine the effect of nutrient enrichment on five ecological services provided by a model coastal system, a shallow seagrass community near Mobile Bay, Alabama (USA): (1) the provision of shelter for fauna; (2) the quality of food provided to first-order consumers; (3) quantity of food provision to first-order consumers and O2/CO2 exchange; (4) producer carbon and nitrogen storage, and (5) water clarity. The results showed a severe negative impact on seagrass density and biomass, which greatly reduced the structural complexity of the community and provision of shelter to fauna. Water clarity and the standing stock of producer carbon were reduced in the fertilized area in comparison with the control area. In contrast, nutrient addition did not affect in any consistent way the total quantity of food available for first-order consumers, the net exchange of O2/CO2, or the standing stock of producer nitrogen in the community. The nutritional quality of the food available for first-order consumers increased with fertilization. These results show that the impacts of nutrient enrichment on the services provided by natural systems may be disparate, ranging from negative to positive. These findings suggest that management policies for anthropogenic eutrophication will depend on the specific ecosystem service targeted. In the case of shallow seagrass beds, the loss of biogenic habitat and drastic impacts on commercially important fauna may be sufficiently alarming to warrant rigorous control of coastal eutrophication. PMID- 21639062 TI - Fresh start. Replacement facility meets expanding regional needs. PMID- 21639063 TI - Converging realities. Integrating medical technology with operations and services. PMID- 21639064 TI - Critical evidence. Exploring research-driven interior design. PMID- 21639065 TI - Rolling it out. Selecting carpet for the health care environment. PMID- 21639066 TI - Code reform initiative continues apace. PMID- 21639067 TI - Constantly increasing membership value. PMID- 21639068 TI - In and out. New guidance on securing egress doors. PMID- 21639069 TI - State your case. Validating staffing needs when making budget requests. PMID- 21639070 TI - Sticker shock: the price of physician assistant education. PMID- 21639071 TI - Controversy and physician assistant education. PMID- 21639072 TI - Evaluation of multiple variables predicting the likelihood of passage and failure of PANCE. AB - PURPOSE: A significant positive correlation between the Physician Assistant National Certifying Examination (PANCE) and the PAEA Physician Assistant Clinical Knowledge Rating and Assessment Tool (PACKRAT) has been found in previous studies. This study's goal was to improve prediction of PANCE failure using other predictive measures, alone and in combination, with the PACKRAT. METHODS: Correlation and discriminate and regression analyses were conducted on 3 years of data (2007-2009) collected from Chatham University PA Studies graduates, with convergent results. RESULTS: Significant positive correlations between the PANCE and each of the assessment measures were found (in order of significance): (1) the PACKRAT exam; (2) a summative multiple choice question exam given near the end of the clinical (second) year; (3) average multiple-choice-question-exam results from the didactic (first) year; (4) prerequisite preadmission GPA; and (5) overall preadmission GPA. Discriminant analyses revealed the combined solution of PACKRAT, SUMM, and MCQ to most accurately differentiate between the "pass" versus "fail" groups on the PANCE. Most importantly, regression analyses revealed the combined solution of SUMM and MCQ to most accurately predict PANCE performance at the low end of the scale. CONCLUSION: This research suggests that all assessment measures tested can provide helpful estimates of PANCE performance; however the combined SUMM and MCQ solution provided the most reliable and accurate prediction of PANCE performance for "high risk" students. Other physician assistant programs could develop analyses of their internal testing assessments to identify students at risk of PANCE failure and allocate institutional and program resources to improve rates of PANCE passage. PMID- 21639073 TI - Physician assistant students' perceptions of an entry-level doctorate degree. AB - PURPOSE: In recent years, a growing trend toward clinical doctorate degrees has emerged in several allied health professions. However, few studies have been conducted within the physician assistant (PA) profession related to changing the entry-level degree for PAs to a clinical doctorate. METHODS: A descriptive, quantitative study assessing PA students' perceptions of changing the entry-level credential for PAs to a clinical doctorate was conducted. Thirty randomly selected programs with 1,966 students were invited to participate in the survey. RESULTS: Of the programs invited, 25 (83%) participated, with 486 (25%) full-time students completing the survey. Of the respondents, 56.1% (272) were first-year students and 43.9% (213) were second-year students. Both groups had a negative perception of changing the entry-level degree for PAs to a clinical doctorate, indicating the doctorate will raise the cost of PA education and discourage some people from entering the profession (82.1%, n = 398); a doctorate for PAs will cause confusion for patients (71.3%, n = 346); and that there is no need for the profession-specific doctorate since the master's degree sufficiently prepares PAs to practice in today's health care setting (70.9%, n = 344). CONCLUSIONS: The present findings lend additional support to previous studies, endorsing the master's degree as the entry-level and terminal degrees. PMID- 21639074 TI - A comparison of behavioral and multiple mini-interview formats in physician assistant program admissions. AB - PURPOSE: The interview remains a widely used tool in health professions program admissions. The purpose of this study was to compare the use of a behavioral interview format with the multiple mini-interview format in measuring desired noncognitive behaviors. METHODS: This dual cohort, observational, comparative study used a polytomous rating-scale model to analyze the results from two homogeneous groups of physician assistant (PA) program applicants (total N = 176). One group (n = 93) participated in two 20-minute behavioral interviews conducted by two raters per interviewee. The behavioral format included questions related to past behaviors and performance as a way to identify latent professionalism characteristics. The second group (n = 83) completed ten separate 7-minute stations with one rater per station. Each of the mini-stations assigned a specific topic and/or task to be completed. The score distributions related to applicant performance and station difficulty were plotted using Rasch analysis software. RESULTS: The behavioral interview format and multiple mini-interview had similar model fit. The behavioral interview did not adequately measure differences in applicant characteristics. In contrast, the multiple mini interview measured more variation in noncognitive traits and identified better matching of station difficulty and person ability. CONCLUSIONS: In this study the multiple mini-interview format was a more reliable admissions tool in detecting latent professionalism attributes among PA program applicants. The multiple mini interview format appeared to measure professional potential and organizational fit better than the behavioral interview format. A larger study across several programs may provide additional support for these findings. PMID- 21639075 TI - The relationship between formative and summative examinations and PANCE scores; can the past predict the future? AB - PURPOSE: This paper describes how a systematic analysis of students at risk for failing the Physician Assistant National Certifying Examination (PANCE) may be used to identify which students may benefit from intervention prior to taking the PANCE and thus increase the likelihood of successful completion of the PANCE. The intervention developed and implemented uses various formative and summative examinations to predict students' PANCE scores with a high degree of accuracy. METHODS: Eight end-of-rotation exams (EOREs) based upon discipline-specific diseases and averaging 100 questions each, a 360-question PANCE simulation (SUMM I), the PACKRAT, and a 700-question summative cognitive examination based upon the NCCPA blueprint (SUMM II) were administered to all students enrolled in the program during the clinical year starting in January 2010 and concluding in December 2010. RESULTS: When the PACKRAT, SUMM I, SUMM II, and the surgery, women's health, and pediatrics EOREs were combined in a regression model, an Rvalue of 0.87 and an R2 of 0.75 were obtained. A predicted score was generated for the class of 2009. The predicted PANCE score based upon this model had a final correlation of 0.790 with the actual PANCE score. CONCLUSION: This pilot study demonstrated that valid predicted scores could be generated from formative and summative examinations to provide valuable feedback and to identify students at risk of failing the PANCE. PMID- 21639076 TI - How social networking sites affect your employment. Interface with facebook and other social media sites may have legal consequences. PMID- 21639077 TI - Evidence-based medicine databases: changing needs along the path from physician assistant student to clinician. PMID- 21639078 TI - Physician assistant education: five countries. AB - BACKGROUND: Physician assistant (PA) education has undergone substantial change since the late 1960s. After four decades of development, other countries have taken a page from the American experience and launched their own instructional initiatives. The diversity in how different countries approach education and produce a PA for their nation's needs provides an opportunity to make comparisons. The intent of this study was to document and describe PA programs in Australia, Canada, the United Kingdom, The Netherlands, and the United States. METHODS: We reviewed the literature and contacted a network of academics in various institutions to obtain primary information. Each contact was asked a set of basic questions about the country, the PA program, and the deployment of graduates. Information on US PA programs was obtained from the Physician Assistant Education Association. RESULTS: At year's end 2010, the following was known about PA development: Australia, one program; Canada, four programs; United Kingdom, four programs; The Netherlands, five programs; the United States, 154 programs. Trends in program per capita growth remain the largest in the United States, followed by The Netherlands and Canada. The shortest program length was 24 months and the longest, 36 months. Outside the United States, almost all programs are situated in an academic health center ([AHC] defined as a medical university, a teaching hospital, and a nursing or allied health school), whereas only one-third of US PA programs are in AHCs. All non-US programs receive public/government funding whereas American programs are predominately private and depend on tuition to fund their programs. CONCLUSION: The PA movement is a global phenomenon. How PAs are being educated, trained, and deployed is known only on the basic level. We identify common characteristics, unique aspects, and trends in PA education across five nations, and set the stage for collaboration and analysis of optimal educational strategies. Additional information is needed on lesser-known PA programs outside these five countries. PMID- 21639079 TI - Richard Reed's chance encounter with the infamous Dr. Van Eeden. PMID- 21639082 TI - Electronic and vibrational transition moment directions in 7-dimethylamino-3 methyl-N-methyl-d3-4-phenylethynylcarbostyril. AB - We report the synthesis and photophysical characterization of 7-dimethylamino-3 methyl-N-methyl-d(3)-4-phenylethynylcarbostyril, a chromophore of interest as a rotator in surface-mounted molecular rotors. Measurement of UV-vis absorption and fluorescence spectra, steady state fluorescence and excitation anisotropy, and linear dichroism in the IR and UV-vis permitted a determination of absolute vibrational and electronic transition moment directions in this previously unreported chromophore. The first singlet-singlet absorption and fluorescence are polarized perpendicular to the axle of the rotator. Density functional theory calculations of electronic excitation and vibrational frequencies gave results in very good agreement with those observed. Calculated IR transition moment directions showed rather poor agreement with experiment. PMID- 21639083 TI - Rotationally resolved C2 symmetric conformers of bis-(4-hydroxyphenyl)methane: prototypical examples of excitonic coupling in the S1 and S2 electronic states. AB - Rotationally resolved microwave and ultraviolet spectra of jet-cooled bis-(4 hydroxyphenyl)methane (b4HPM) have been obtained using Fourier-transform microwave and UV laser/molecular beam spectrometers. A recent vibronic level study of b4HPM [Rodrigo, C. P.; Muller, C. W.; Pillsbury, N. R.; James, W. H., III; Plusquellic, D. F.; Zwier, T. S. J. Chem. Phys. 2011, 134, 164312] has assigned two conformers distinguished by the orientation of the in-plane OH groups and has identified two excitonic origins in each conformer. In the present study, the rotationally resolved bands of all four states have been well-fit to asymmetric rotor Hamiltonians. For the lower exciton (S(1)) levels, the transition dipole moment (TDM) orientations are perpendicular to the C(2) symmetry axes and consist of 41(2):59(2) and 34(2):66(2)% a:c hybrid-type character. The S(1) levels are therefore delocalized states of B symmetry and represent the antisymmetric combinations of the zero-order locally excited states of the p-cresol-like chromophores. The TDM polarizations of bands located at ~132 cm(-1) above the S(1) origins are exclusively b-type and identify them as the upper exciton S(2) origin levels of A symmetry. The TDM orientations and the relative band strengths from the vibronic study have been analyzed within a dipole-dipole coupling model in terms of the localized TDM orientations, MU(loc), on the two chromophores. The out-of-the-ring plane angles of MU(loc) are both near 20 degrees and are similar to results for diphenylmethane [Stearns, J. A.; Pillsbury, N. R.; Douglass, K. O.; Muller, C. W.; Zwier, T. S.; Plusquellic, D. F. J. Chem. Phys. 2008, 129, 224305]. The in-plane angles are, however, rotated by 14 and 18 degrees relative to DPM and, in part, explain the smaller than expected exciton splittings of these two conformers. PMID- 21639084 TI - Ab initio study of ternary complexes X:(HCNH)(+):Z with X, Z = NCH, CNH, FH, ClH, and FCl: diminutive cooperative effects on structures, binding energies, and spin spin coupling constants across hydrogen bonds. AB - Ab initio calculations have been performed on a series of complexes in which (HCNH)(+) is the proton donor and CNH, NCH, FH, ClH, and FCl (molecules X and Z) are the proton acceptors in binary complexes X:HCNH(+) and HCNH(+):Z, and ternary complexes X:HCNH(+):Z. These complexes are stabilized by C-H(+)...A and N H(+)...A hydrogen bonds, where A is the electron-pair donor atom of molecules X and Z. Binding energies of the ternary complexes are less than the sum of the binding energies of the corresponding binary complexes. In general, as the binding energy of the binary complex increases, the diminutive cooperative effect increases. The structures of these complexes, data from the AIM analyses, and coupling constants (1)J(N-H), (1h)J(H-A), and (2h)J(N-A) for the N-H(+)...A hydrogen bonds, and (1)J(C-H), (1h)J(H-A), and (2h)J(C-A) for the C-H(+)...A hydrogen bonds provide convincing evidence of diminutive cooperative effects in these ternary complexes. In particular, the symmetric N...H(+)...N hydrogen bond in HCNH(+):NCH looses proton-shared character in the ternary complexes X:HCNH(+):NCH, while the proton-shared character of the C...H(+)...C hydrogen bond in HNC:HCNH(+) decreases in the ternary complexes HNC:HCNH(+):Z and eventually becomes a traditional hydrogen bond as the strength of the HCNH(+)...Z interaction increases. PMID- 21639085 TI - Polybrominated diphenyl ethers and polychlorinated dibenzo-p-dioxins and dibenzofurans in surface dust at an E-waste processing site in Southeast China. AB - Surface dust collected from printed circuit board recycling workshop floors, roads, a schoolyard, and an outdoor food market in Guiyu, China, a village intensely involved in e-waste processing, were investigated for levels of polybrominated diphenyl ethers (PBDEs) and polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs). PBDE concentrations in dust from workshop-floors (14,800 +/- 5130 ng/g) and on adjacent roads to the workshops (24,900 +/- 31,600 ng/g) were highest among the study sites whereas PCDD/F concentrations were highest at the schoolyard (1316 pg/g) and in a workshop (1264 pg/g). Analyses of <2 mm and <53 MUm dust particle sizes did not show any significant differences in PBDE concentrations. The cytotoxicity was investigated using two bioassays: 7 ethoxyresorufin O-deethylase (EROD-TEQ) and MTT. EROD-TEQ values ranged from 260 to 432 pg/g, with the highest in dust collected from a street lined with workshops. Using the MTT assay, cytoxicity of dust from the plastic chips drying district in Guiyu was higher than dust from the other sites investigated. This study showed that the primitive recycling of e-waste introduced toxic pollutants into the environment which are potentially harmful to the health of e-waste workers and local residents, especially children, and warrants an urgent investigation into POPs related health impacts. PMID- 21639086 TI - Accurate values of some thermodynamic properties for carbon dioxide, ethane, propane, and some binary mixtures. AB - Quasicontinuous PrhoT data of CO(2), ethane, propane, and the [CO(2) + ethane] mixture have been determined along subcritical, critical, and supercritical regions. These data have been used to develop the optimal experimental method and to determine the precision of the results obtained when using an Anton Paar DMA HPM vibrating-tube densimeter. A comparison with data from reference EoS and other authors confirm the quality of our experimental setup, its calibration, and testing. For pure compounds, the value of the mean relative deviation is MRD(rho) = 0.05% for the liquid phase and for the extended critical and supercritical region. For binary mixtures the mean relative deviation is MRD(rho) = 0.70% in the range up to 20 MPa and MRD(rho) = 0.20% in the range up to 70 MPa. The number of experimental points measured and their just quality have enable us to determine some derivated properties with satisfactory precision; isothermal compressibilities, kappa(T), have been calculated for CO(2) and ethane (MRD(kappa(T)) = 1.5%), isobaric expasion coefficients, alpha(P), and internal pressures, pi(i), for CO(2) (MRD(alpha(P)) = 5% and MRD(pi(i)) = 7%) and ethane (MRD(alpha(P)) = 7.5% and MRD(pi(i)) = 8%). An in-depth discussion is presented on the behavior of the properties obtained along subcritical, critical, and supercritical regions. In addition, PuT values have been determined for water and compressed ethane from 273.19 to 463.26 K up to pressures of 190.0 MPa, using a device based on a 5 MHz pulsed ultrasonic system (MRD(u) = 0.1%). With these data we have calibrated the apparatus and have verified the adequacy of the operation with normal liquids as well as with some compressed gases. From density and speed of sound data of ethane, isentropic compressibilities, kappa(s), have been obtained, and from these and our values for kappa(T) and alpha(P), isobaric heat capacities, C(p), have been calculated with MRD(C(p)) = 3%, wich is within that of the EoS. PMID- 21639087 TI - Fluorescent ferritin nanoparticles and application to the aptamer sensor. AB - We synthesized fluorescent ferritin nanoparticles (FFNPs) through bacterial expression of the hybrid gene consisting of human ferritin heavy chain (hFTN-H), spacer (glycine-rich peptide), and enhanced green (or red) fluorescent protein [eGFP (or DsRed)] genes. The self-assembly activity of hFTN-H that leads to the formation of nanoparticles (12 nm in diameter), the conformational flexibility of the C-terminus of hFTN-H, and the glycine-rich spacer enabled eGFPs (or DsReds) to be well displayed on the surface of each ferritin nanoparticle, resulting in the construction of green (or red) FFNPs [gFFNPs (or rFFNPs)]. As compared to eGFP (or DsRed) alone, it is notable that the developed FFNPs showed significantly amplified fluorescence intensity and also enhanced stability. DNA aptamers were chemically conjugated to gFFNP via each eGFP's cysteine residue that was newly introduced through site-directed mutagenesis (Ser175Cys). The DNA aptamer-conjugated gFFNPs were used as a fluorescent reporter probe in the aptamer-based "sandwich" assay of a cancer marker [i.e., platelet-derived growth factor B-chain homodimer (PDGF-BB)] in phosphate-buffered saline buffer or diluted human serum. This is a simple two-step assay without any additional steps for signal amplification, showing that compared to the same aptamer-based assays using eGFP alone or Cy3, the detection signals, affinity of the reporter probe to the cancer marker, and assay sensitivity were significantly enhanced; i.e., the limit of detection was lowered to the 100 fM level. Although the PDGF-BB assay is reported here as a proof-of-concept, the developed FFNPs can be applied in general to any aptamer-based sandwich assays. PMID- 21639088 TI - Matrix sublimation/recrystallization for imaging proteins by mass spectrometry at high spatial resolution. AB - We have employed matrix deposition by sublimation for protein image analysis on tissue sections using a hydration/recrystallization process that produces high quality MALDI mass spectra and high-spatial-resolution ion images. We systematically investigated different washing protocols, the effect of tissue section thickness, the amount of sublimated matrix per unit area, and different recrystallization conditions. The results show that an organic solvent rinse followed by ethanol/water rinses substantially increased sensitivity for the detection of proteins. Both the thickness of the tissue section and the amount of sinapinic acid sublimated per unit area have optimal ranges for maximal protein signal intensity. Ion images of mouse and rat brain sections at 50, 20, and 10 MUm spatial resolution are presented and are correlated with hematoxylin and eosin (H&E)-stained optical images. For targeted analysis, histology-directed imaging can be performed using this protocol where MS analysis and H&E staining are performed on the same section. PMID- 21639089 TI - Anisotropic self-assembling micelles prepared by the direct dissolution of PLA/PEG block copolymers with a high PEG fraction. AB - A series of polylactide-poly(ethylene glycol) (PLA-PEG) block copolymers with a high PEG fraction were synthesized by the ring-opening polymerization of L- or D lactide in the presence of mono- or dihydroxyl PEG using nontoxic zinc lactate as a catalyst. Micelles were then prepared by direct dissolution of the obtained water-soluble copolymers in an aqueous medium without heating or using any organic solvents. Large anisotropic micelles instead of conventional spherical ones were observed from a transmission electron microscopy examination. Various parameters influencing the structure of the novel micelles were considered, such as the copolymer chain structure, molar mass, PEG fraction, copolymer concentration, and stereocomplexation between L- and D-PLA blocks. Anisotropic micelles were obtained for both diblock and triblock copolymers but vanished with increasing molar mass of the copolymers. The morphology of micelles strongly depends on the PEG fraction. Anisotropic micelles were found only in an intermediate EO/LA ratio range in which a higher PEG fraction leads to a higher length/width ratio of micelles. Stereocomplexation between L- and D-PLA or a lower concentration disfavors the formation of anisotropic micelles. Under appropriate concentrations, spherical and anisotropic micelles coexist in the same micellar solution. Moreover, it was found that anisotropic micelles are susceptible to further self-assemble into more organized complex aggregates. Similar results were obtained from light scattering and aqueous gel permeation chromatography measurements. A novel model is proposed to explain the formation of anisotropic micelles and the effects of various parameters on the structure of micelles in an aqueous medium. PMID- 21639090 TI - Magneto-controlled graphene immunosensing platform for simultaneous multiplexed electrochemical immunoassay using distinguishable signal tags. AB - A novel flow-through multiplexed immunoassay protocol for simultaneous electrochemical determination of carcinoembryonic (CEA) and alpha-fetoprotein (AFP) in biological fluids was designed using biofunctionalized magnetic graphene nanosheets (MGO) as immunosensing probes and multifunctional nanogold hollow microspheres (GHS) as distinguishable signal tags. The probes were fabricated by means of co-immobilization of primary anti-CEA (Ab(1)) and anti-AFP (Ab(2)) antibodies on the Fe(3)O(4) nanoparticle-coated graphene nanosheets (MGO Ab(1,2)). The reverse-micelle method was used for the synthesis of distinguishable signal tags by encapsulation of horseradish peroxide (HRP) thionine and HRP-ferrocene into nanogold hollow microspheres, respectively, which were utilized as labels of the corresponding GHS-Ab(1) and GHS-Ab(2). A sandwich type immunoassay format was employed for the online detection of CEA and AFP by coupling a flow-through detection cell with an external magnet. The assay was based on the catalytic reduction of H(2)O(2) at the various peak potentials in the presence of the corresponding mediators. Experimental results revealed that the multiplexed electrochemical immunoassay enabled the simultaneous monitoring of AFP and CEA in a single run with wide working ranges of 0.01-200 ng mL(-1) for AFP and 0.01-80 ng mL(-1) for CEA. The detection limits (LODs) for both analytes at 1.0 pg mL(-1) (at 3s(B)) were very low. No obvious nonspecific adsorption and cross-talk were observed during a series of analyses to detect target analytes. Intraassay and interassay coefficients of variation were <10%. Importantly, the methodology was evaluated for the analysis of clinical serum specimens, receiving a good correlation between the flow-through multiplexed electrochemical immunoassay and an electrochemiluminescence method as a reference. PMID- 21639091 TI - Phosphorylation, but not alternative splicing or proteolytic degradation, is conserved in human and mouse cardiac troponin T. AB - Cardiac troponin T (cTnT), the tropomyosin binding subunit of the troponin complex, plays a pivotal regulatory role in the Ca(2+)-mediated interaction between actin thin filament and myosin thick filament. The post-translational modifications (PTMs) and alternative splicing of cTnT may represent important regulatory mechanisms of cardiac contractility. However, a complete characterization of PTMs and alternatively spliced isoforms in cTnT present in vivo is lacking. Top-down protein mass spectrometry (MS) analyzes whole proteins, thus providing a global view of all types of modifications, including PTMs and sequence variants, simultaneously in one spectrum without a priori knowledge. In this study, we applied an integrated immunoaffinity chromatography and top-down MS approach to comprehensively characterize PTMs and alternatively spliced isoforms of cTnT purified from healthy human and wild-type mouse heart tissue. High-resolution Fourier transform MS revealed that human cTnT (hcTnT) and mouse cTnT (mcTnT) have similar phosphorylation patterns, whereas higher molecular heterogeneity was observed for mcTnT than hcTnT. Further MS/MS fragmentation of monophosphorylated hcTnT and mcTnT by electron capture dissociation and collisionally activated dissociation unambiguously identified Ser1 as the conserved in vivo phosphorylation site. In contrast, we identified a single spliced isoform for hcTnT but three alternatively spliced isoforms for mcTnT. Moreover, we observed distinct proteolytic degradation products for hcTnT and mcTnT. This study also demonstrates the advantage of top-down MS/MS with complementary fragmentation techniques for the identification of modification sites in the highly acidic N-terminal region of cTnT. PMID- 21639092 TI - Antioxidant and cytotoxic activities of carob tree fruit pulps are strongly influenced by gender and cultivar. AB - Extracts from fruit pulps of six female cultivars and two hermaphrodite Portuguese carob trees [(Ceratonia siliqua L., Fabaceae)] exhibited strong antioxidant activity and were rich in phenolic compounds. The extracts decreased the viability of different human cancer cell lines on a dose- and time-dependent manner. Gender and cultivar significantly influenced the chemical content and the biological activities of the extracts. Extracts from hermaphrodite trees had a higher content of phenolic compounds, and exhibited higher antioxidant and cytotoxic activities. Among females, cv. Aida had the highest radical scavenging activity and total content of phenolics, Mulata the highest capacity to inhibit lipid oxidation and Gasparinha the strongest cytotoxic activity on HeLa cells. The decrease in cell viability was associated with apoptosis on HeLa and MDA-MB 231 lines. (+)-Catechin and gallic acid (GA) were the main compounds identified in the extracts, and GA contributed to the antioxidant activity. Our results show that the antioxidant and cytotoxic activities of carob tree fruit pulps are strongly influenced by gender and cultivar, and provide new knowledge about the advantages of hermaphrodite trees over female cultivars, namely, as a source of compounds with biological interest, which may represent an increase of their agronomic interest. PMID- 21639093 TI - Nutrient and trace-metal removal by Bauxsol pellets in wastewater treatment. AB - In this study, Bauxsol pellets packed in PVC columns were used to remove nutrients and trace-metals from municipal wastewater during a 6 months field trial. Bauxsol pellet columns showed a high phosphate removal rate via precipitation of PO(4)(3-) with Ca(2+) and Mg(2+) ions: at 90% in the 1st month; at 80% from the second to fifth months; and at 60% in the sixth month. Pellet bound total phosphorus and Colwell phosphorus were 7.3 g/kg and 2 g/kg and are about 20 times the concentrations found in most fertile soils. Trace-metals in effluents were bound, probably irreversibly under the columns' environmental conditions, to the Bauxsol minerals that have high surface area to volume ratios and high charge to mass ratios. Experimental results showed a complex nitrogen cycle operating within the Bauxsol pellet columns including anoxic nitrification, denitrification, and anammox processes. Although a transient pH spike, associated with the release of unreacted CaO from the cement binder used in the pellets, was observed, this may be readily corrected through post-treatment pH adjustment. Hence, the geochemistry of Bauxsol pellets can effectively remove and bind nutrients and trace-metals during wastewater treatment, and further research may show that saturated spent pellets can be used as fertilizer. PMID- 21639094 TI - Interfacial activity of metal beta-diketonato complexes: in situ generation of amphiphiles by water coordination. AB - Symmetric transition metal complexes of 2,4-pentanedione (acetyl acetone) are interfacially active: Spinning drop tensiometry reveals lowering of the interfacial tension at the water-organic interface, most pronounced for [Cr(acac)(3)], [Fe(acac)(3)], [Zr(acac)(4)], and [Hf(acac)(4)]. The interfacial activity is explained by the in situ generation of amphiphilic species. Based on tensiometry and (1)H and diffusion-ordered NMR spectroscopy (DOSY NMR), hydrogen bonding of the organically dissolved complexes with water, in some cases in combination with inner-sphere hydrolytic coordination, is identified as the primary origin of this amphiphilicity. The complexes are a rare example of symmetric molecules that turn amphiphilic only upon interfacial interaction with water. PMID- 21639095 TI - Reversible and selective O2 chemisorption in a porous metal-organic host material. AB - The metal-organic host material [{Co(III)(2)(bpbp)(O(2))}(2)bdc](PF(6))(4) (1.2O(2); bpbp(-) = 2,6-bis(N,N-bis(2-pyridylmethyl)aminomethyl)-4-tert butylphenolato; bdc(2-) = 1,4-benzenedicarboxylato) displays reversible chemisorptive desorption and resorption of dioxygen through conversion to the deoxygenated Co(II) form [{Co(II)(2)(bpbp)}(2)bdc](PF(6))(4) (1). Single crystal X-ray diffraction analysis indicates that the host lattice 1.2O(2), achieved through desorption of included water guests from the as-synthesized phase 1.2O(2).3H(2)O, consists of an ionic lattice containing discrete tetranuclear complexes, between which lie void regions that allow the migration of dioxygen and other guests. Powder X-ray diffraction analyses indicate that the host material retains crystallinity through the dioxygen desorption/chemisorption processes. Dioxygen chemisorption measurements on 1 show near-stoichiometric uptake of dioxygen at 5 mbar and 25 degrees C, and this capacity is largely retained at temperatures above 100 degrees C. Gas adsorption isotherms of major atmospheric gases on both 1 and 1.2O(2) indicate the potential suitability of this material for air separation, with a O(2)/N(2) selectivity factor of 38 at 1 atm. Comparison of oxygen binding in solution and in the solid state indicates a dramatic increase in binding affinity to the complex when it is incorporated in a porous solid. PMID- 21639096 TI - A general copper-BINAP-catalyzed asymmetric propargylation of ketones with propargyl boronates. AB - An operationally simple copper-BINAP-catalyzed, highly enantioselective propargylation of ketones is presented. The methodology was developed as an enantioselective process for methyl ethyl ketone and shown to be applicable to a wide variety of prochiral ketones. The resulting homopropargyl adducts are versatile latent carbonyls from which gamma-butyrolactones, beta-hydroxy methyl ketones, and beta-hydroxycarboxylates are readily obtained. PMID- 21639097 TI - Highly efficient visible-light-driven photocatalytic hydrogen production of CdS cluster-decorated graphene nanosheets. AB - The production of clean and renewable hydrogen through water splitting using photocatalysts has received much attention due to the increasing global energy crises. In this study, a high efficiency of the photocatalytic H(2) production was achieved using graphene nanosheets decorated with CdS clusters as visible light-driven photocatalysts. The materials were prepared by a solvothermal method in which graphene oxide (GO) served as the support and cadmium acetate (Cd(Ac)(2)) as the CdS precursor. These nanosized composites reach a high H(2) production rate of 1.12 mmol h(-1) (about 4.87 times higher than that of pure CdS nanoparticles) at graphene content of 1.0 wt % and Pt 0.5 wt % under visible light irradiation and an apparent quantum efficiency (QE) of 22.5% at wavelength of 420 nm. This high photocatalytic H(2)-production activity is attributed predominantly to the presence of graphene, which serves as an electron collector and transporter to efficiently lengthen the lifetime of the photogenerated charge carriers from CdS nanoparticles. This work highlights the potential application of graphene-based materials in the field of energy conversion. PMID- 21639098 TI - Fourier transform infrared and Raman spectroscopies for the rapid detection, enumeration, and growth interaction of the bacteria Staphylococcus aureus and Lactococcus lactis ssp. cremoris in milk. AB - Staphylococcus aureus is one of the main pathogenic microorganisms found in milk and dairy products and has been involved in bacterial foodborne outbreaks in the past. Current enumeration techniques for bacteria are very time-consuming, typically taking 24 h or longer, and bacterial antagonism in the form of lactic acid bacteria (LAB) may inhibit the growth of S. aureus . Therefore, the aim of this investigation was to establish the accuracy and sensitivity of rapid nondestructive metabolic fingerprinting techniques, such as Fourier transform infrared (FT-IR) spectroscopy and Raman spectroscopy (RS), in combination with multivariate analysis techniques, for the detection and enumeration of S. aureus in milk, as well as to study the growth interaction between S. aureus and Lactococcus lactis ssp. cremoris , a common LAB. The two bacterial species were investigated both in a pure monoculture and in a combined inoculated coculture after inoculation into ultraheated milk during the first 24 h of growth at 37 degrees C. Plating techniques were used to obtain primary reference data for viable bacteria counts. Principal component discriminant function analysis, canonical correlation analysis, partial least-squares (PLS), and kernel PLS (KPLS) multivariate statistical techniques were employed to analyze the data. FT IR provided very reasonable quantification results both with PLS and KPLS, the latter providing marginally better predictions, with correlation coefficients in the test set (Q(2)) and training set (R(2)) varying from 0.64 to 0.76 and from 0.78 to 0.88 for different bacterial sample combinations. RS results were less encouraging with high degrees of error and poor correlation to viable bacterial counts. S. aureus growth was not inhibited by the presence of the LAB, but metabolic fingerprinting of the coculture indicated that the phenotype of this dual bacterial culture was closer to that of pure LAB cultures. In conclusion, FT IR spectroscopy in combination with the above multivariate techniques appears to be a promising discrimination and enumeration analytical technique for the two bacterial species. In addition, it has been demonstrated that the L. cremoris metabolic effect in milk dominates that of S. aureus even though there was no growth antagonism observed. PMID- 21639099 TI - Microscopic wetting of self-assembled monolayers with different surfaces: a combined molecular dynamics and quantum mechanics study. AB - Molecular dynamics simulations are used to study the micronature of the organization of water molecules on the flat surface of well-ordered self assembled monolayers (SAMs) of 18-carbon alkanethiolate chains bound to a silicon (111) substrate. Six different headgroups (-CH(3), -C?C, -OCH(3), -CN, -NH(2), COOH) are used to tune the character of the surface from hydrophobic to hydrophilic, while the level of hydration is consistent on all six SAM surfaces. Quantum mechanics calculations are employed to optimize each alkyl chain of the different SAMs with one water molecule and to investigate changes in the configuration of each headgroup under hydration. We report the changes of the structure of the six SAMs with different surfaces in the presence of water, and the area of the wetted surface of each SAM, depending on the terminal group. Our results suggest that a corrugated and hydrophobic surface will be formed if the headgroups of SAM surface are not able to form H-bonds either with water molecules or between adjacent groups. In contrast, the formation of hydrogen bonds not only among polar heads but also between polar heads and water may enhance the SAM surface hydrophilicity and corrugation. We explicitly discuss the micromechanisms for the hydration of three hydrophilic SAM (CN-, NH(2)- and COOH terminated) surfaces, which is helpful to superhydrophilic surface design of SAM in biomimetic materials. PMID- 21639100 TI - Photoisomerization of stilbene: a spin-flip density functional theory approach. AB - The photoisomerization process of 1,2-diphenylethylene (stilbene) is investigated using the spin-flip density functional theory (SFDFT), which has recently been shown to be a promising approach for locating conical intersection (CI) points (Minezawa, N.; Gordon, M. S. J. Phys. Chem. A2009, 113, 12749). The SFDFT method gives valuable insight into twisted stilbene to which the linear response time dependent DFT approach cannot be applied. In contrast to the previous SFDFT study of ethylene, a distinct twisted minimum is found for stilbene. The optimized structure has a sizable pyramidalization angle and strong ionic character, indicating that a purely twisted geometry is not a true minimum. In addition, the SFDFT approach can successfully locate two CI points: the twisted-pyramidalized CI that is similar to the ethylene counterpart and another CI that possibly lies on the cyclization pathway of cis-stilbene. The mechanisms of the cis--trans isomerization reaction are discussed on the basis of the two-dimensional potential energy surface along the twisting and pyramidalization angles. PMID- 21639101 TI - Mimics of peptide turn backbone and side-chain geometry by a general approach for modifying azabicyclo[5.3.0]alkanone amino acids. AB - Peptide mimics with constrained backbone and side-chain geometry are important tools for studying structure activity relationships of biologically active candidates. A general method for creating beta-turn mimics possessing side-chain diversity has been developed featuring diastereoselective S(N)1 displacements of an iodide precursor. In particular, 6-iodo-pyrroloazepin-2-one amino ester 10 has served as a common precursor in reactions with a variety of alcohol, phenol, nitrate, and azide nucleophiles to provide an array of constrained peptide mimics. PMID- 21639102 TI - Total synthesis of the cyanolide A aglycon. AB - The synthesis of the potent molluscicide cyanolide A has been achieved in 10 steps without the use of protecting groups. The synthesis features a key Sakurai macrocyclization/dimerization reaction that simultaneously forms both tetrahydropyran rings and the macrocycle of the natural product. PMID- 21639103 TI - Solvent mixture induced self assembly of a terthiophene based rod-coil block co oligomer. AB - We describe the synthesis and self-assembly of a terthiophene-oligostyrene based block co-oligomer Ter-OSa. While the self-assembly of block copolymers is normally achieved using block-selective solvents, tetrahydrofuran (THF) is a good solvent for both terthiophene and oligostyrene blocks. The self-assembly in this case arises with the nonsolvent (H(2)O) in THF/H(2)O mixtures. The absorption spectra showed a blue shift and fluorescence spectra showed quenching of fluorescence with increasing water content indicating that the Ter-OSa self assembled to form H-aggregates. The morphology was studied using scanning electron microscopy. Depending upon the amount of water the Ter-OSa undergoes morphological transition from hollow to rigid spheres. However at higher water content the Ter-OSa formed large spheres from the fusion of smaller spheres. A mechanism of morphological transition from hollow spheres to rigid spheres is proposed. PMID- 21639104 TI - Graphene-DNAzyme based biosensor for amplified fluorescence "turn-on" detection of Pb2+ with a high selectivity. AB - On the basis of the remarkable difference in affinity of graphene (GO) with ssDNA containing a different number of bases in length, we for the first time report a GO-DNAzyme based biosensor for amplified fluorescence "turn-on" detection of Pb(2+). A FAM-labeled DNAzyme-substrate hybrid acted as both a molecular recognition module and signal reporter and GO as a superquencher. By taking advantage of the super fluorescence quenching efficiency of GO, our proposed biosensor exhibits a high sensitivity toward the target with a detection limit of 300 pM for Pb(2+), which is lower than previously reported for catalytic beacons. Moreover, with the choice of a classic Pb(2+)-dependent GR-5 DNAzyme instead of 8 17 DNAzyme as the catalytic unit, the newly designed sensing system also shows an obviously improved selectivity than previously reported methods. Moreover, the sensing system was used for the determination of Pb(2+) in river water samples with satisfying results. PMID- 21639105 TI - Electric-field-induced changes in absorption and fluorescence of the green fluorescent protein chromophore in a PMMA film. AB - External electric field effects on absorption, fluorescence, and fluorescence decay of p-HBDI that is a model compound of the chromophore of GFP have been examined in a poly(methyl methacrylate) film. The electroabsorption spectrum is similar in shape to the first derivative of the absorption spectrum, which results from the difference in molecular polarizability between the ground state and the Franck-Condon excited state. The electrophotoluminescence spectrum is dominated by the corresponding fluorescence spectrum, indicating the enhancement of the fluorescence intensity in the presence of external electric fields. The direct measurements of the electric field effect on the fluorescence decay profile suggest that the field-induced deceleration of the nonradiative process contributes to the increase in the fluorescence intensity in the presence of electric fields. PMID- 21639106 TI - High-throughput screening of small molecule libraries using SAMDI mass spectrometry. AB - High-throughput screening is a common strategy used to identify compounds that modulate biochemical activities, but many approaches depend on cumbersome fluorescent reporters or antibodies and often produce false-positive hits. The development of "label-free" assays addresses many of these limitations, but current approaches still lack the throughput needed for applications in drug discovery. This paper describes a high-throughput, label-free assay that combines self-assembled monolayers with mass spectrometry, in a technique called SAMDI, as a tool for screening libraries of 100,000 compounds in one day. This method is fast, has high discrimination, and is amenable to a broad range of chemical and biological applications. PMID- 21639107 TI - Dinuclear Co(II)Co(III) mixed-valence and Co(III)Co(III) complexes with N- and O donor ligands: characterization and water oxidation studies. AB - The tridentate ligand N-methyl-N,N-bis(2-pyridylmethyl)amine (L) has been employed to synthesize a dinuclear Co(II)Co(III) mixed-valence complex containing MU-methoxo and MU-carboxylato bridging ligands, [LCo(II)(MU-carboxylato)bis(MU methoxo)Co(III)L](ClO(4))(2). In this complex, the two pseudo-octahedral Co centers have an identical ligand environment, yet the average Co-N and Co-O bond distances at the two Co ions differ significantly. Electrochemical, spectroscopic, and magnetic susceptibility measurements confirm that it belongs to a localized Class II mixed-valence system, despite the presence of a short Co...Co distance of 3.021 A. Oxidation of this Co(II)Co(III) complex leads to formation of the corresponding Co(III)Co(III) complex that was characterized structurally and spectroscopically. In addition, dinuclear and trinuclear MU hydroxo Co(III) complexes have been obtained in the presence of phosphate anions and absence of methanol, respectively, suggesting that an additional bridging ligand is needed to stabilize the Co(III)bis(MU-hydroxo)Co(III) fragment. Moreover, the ability of the mixed-valence Co(II)Co(III) complex and the three related Co(III) complexes to electrocatalytically oxidize water was also investigated. The observed limited water oxidation catalytic ability for these systems suggests that a multinuclear Co cluster and/or presence of O-rich ligands may be needed for the generation of efficient molecular Co-based water oxidation catalysts. PMID- 21639108 TI - Cs2USi6O15: a tetravalent uranium silicate. AB - A uranium(IV) silicate has been synthesized under high-temperature, high-pressure hydrothermal conditions. The structure consists of unbranched dreier single layers with the composition [Si(2)O(5)] that are connected by UO(6) octahedra to form a 3D framework with 7-ring channels where the Cs(+) cations are located. Each UO(6) octahedron spans four neighboring dreier single chains and, therefore, introduces a high degree of corrugation in the silicate layers. The U 4f X-ray photoelectron spectroscopy spectrum was measured to confirm the valence state of the uranium. A comparison of related metal silicate structures is made. After the synthesis of this compound, all members in the family of uranium silicates and germanates with oxidation states of uranium from 4+ to 6+ have been observed. PMID- 21639110 TI - One-step, nonenzymatic synthesis of O-acetyl-ADP-ribose and analogues from NAD and carboxylates. AB - O-Acetyl-ADP-ribose (OAADPR) is a metabolite produced from nicotinamide adenine dinucleotide (NAD) as a product of sirtuin-mediated protein deacetylation. We present here a simple, one-step, nonenzymatic synthesis of OAADPR from NAD and sodium acetate in acetic acid. We extended the reaction to other carboxylic acids, demonstrating that the reaction between NAD and nonaqueous carboxylate buffers produces mixtures of the corresponding 2'- and 3'-carboxylic esters. PMID- 21639109 TI - Chemoselectivity in chemical biology: acyl transfer reactions with sulfur and selenium. AB - A critical source of insight into biological function is derived from the chemist's ability to create new covalent bonds between molecules, whether they are endogenous or exogenous to a biological system. A daunting impediment to selective bond formation, however, is the myriad of reactive functionalities present in biological milieu. The high reactivity of the most abundant molecule in biology, water, makes the challenges all the more difficult. We have met these challenges by exploiting the reactivity of sulfur and selenium in acyl transfer reactions. The reactivity of both sulfur and selenium is high compared with that of their chalcogen congener, oxygen. In this Account, we highlight recent developments in this arena, emphasizing contributions from our laboratory. One focus of our research is furthering the chemistry of native chemical ligation (NCL) and expressed protein ligation (EPL), two related processes that enable the synthesis and semisynthesis of proteins. These techniques exploit the lower pK(a) of thiols and selenols relative to alcohols. Although a deprotonated hydroxyl group in the side chain of a serine residue is exceedingly rare in a biological context, the pK(a) values of the thiol in cysteine (8.5) and of the selenol in selenocysteine (5.7) often render these side chains anionic under physiological conditions. NCL and EPL take advantage of the high nucleophilicity of the thiolate as well as its utility as a leaving group, and we have expanded the scope of these methods to include selenocysteine. Although the genetic code limits the components of natural proteins to 20 or so alpha-amino acids, NCL and EPL enable the semisynthetic incorporation of a limitless variety of nonnatural modules into proteins. These modules are enabling chemical biologists to interrogate protein structure and function with unprecedented precision. We are also pursuing the further development of the traceless Staudinger ligation, through which a phosphinothioester and azide form an amide. We first reported this chemical ligation method, which leaves no residual atoms in the product, in 2000. Our progress in effecting the reaction in water, without an organic cosolvent, was an important step in the expansion of its utility. Moreover, we have developed the traceless Staudinger reaction as a means for immobilizing proteins on a solid support, providing a general method of fabricating microarrays that display proteins in a uniform orientation. Along with NCL and EPL, the traceless Staudinger ligation has made proteins more readily accessible targets for chemical synthesis and semisynthesis. The underlying acyl transfer reactions with sulfur and selenium provide an efficient means to synthesize, remodel, and immobilize proteins, and they have enabled us to interrogate biological systems. PMID- 21639111 TI - Polymer brushes on graphene. AB - A critical bottleneck for the widespread use of single layer graphene is the absence of a facile method of chemical modification which does not diminish the outstanding properties of the two-dimensional sp(2) network. Here, we report on the direct chemical modification of graphene by photopolymerization with styrene. We demonstrate that photopolymerization occurs at existing defect sites and that there is no detectable disruption of the basal plane conjugation of graphene. This method thus offers a route to define graphene functionality without degrading its electronic properties. Furthermore, we show that photopolymerization with styrene results in self-organized intercalative growth and delamination of few layer graphene. Under these reaction conditions, we find that a range of other vinyl monomers exhibits no reactivity with graphene. However, we demonstrate an alternative route by which the surface reactivity can be precisely tuned, and these monomers can be locally grafted via electron-beam induced carbon deposition on the graphene surface. PMID- 21639112 TI - Finding common ground. PMID- 21639113 TI - Photodegradation of the herbicide imazethapyr in aqueous solution: effects of wavelength, pH, and natural organic matter (NOM) and analysis of photoproducts. AB - The photodegradation of imazethapyr, 5-ethyl-2-(4-isopropyl-4-methyl-5-oxo-4,5 dihydroimidazol-1H-3-yl)nicotinic acid, has been investigated in phosphate buffers and in buffered solutions containing natural organic matter (NOM). Imazethapyr degrades most quickly under 253.7 nm light and at pH values >4. The presence of NOM in solution caused the reaction rate constants for the photodegradation to decrease, with higher concentrations of NOM having a larger effect. Calculations suggest light screening is the major effect of the NOM. Seven photoproducts have been identified, and a photodegradation mechanism is proposed. PMID- 21639114 TI - Primary amino acid derivatives: compounds with anticonvulsant and neuropathic pain protection activities. AB - Pharmacological management remains the primary method to treat epilepsy and neuropathic pain. We have advanced a novel class of anticonvulsants termed functionalized amino acids (FAAs). In this study, we examine FAA derivatives from which the terminal acetyl moiety was removed and termed these compounds primary amino acid derivatives (PAADs). Twenty-seven PAADs were prepared; the central C(2) R-substituent was varied, including C(2) stereochemistry, and the compounds were tested in rodent models of seizures and neuropathic pain. C(2)-Hydrocarbon N benzylamide PAADs were potent anticonvulsants and excellent anticonvulsant activity (mice, ip; rat, po) was observed for C(2) R-substituted PAADs in which the R group was ethyl, isopropyl, or tert-butyl, and the C(2) stereochemistry conformed to the d-amino acid configuration ((R)-stereoisomer). These values surpassed the activities of several clinical antiepileptic drugs. The C(2) (R) ethyl and C(2) (R)-isopropyl PAADs also displayed excellent activities in the mouse (ip) formalin neuropathic pain model. Significantly, unlike the FAA structure-activity relationship, PAAD anticonvulsant activity increased upon substitution of a methylene unit for a heteroatom in the R-substituent that was one atom removed from the C(2) site, suggesting that these PAADs function by a different pathway than FAAs. PMID- 21639115 TI - Acrylodan-labeled smooth muscle tropomyosin reports differences in the effects of troponin and caldesmon in the transition from the active state to the inactive state. AB - Changes in the orientation of tropomyosin on actin are important for the regulation of striated muscle contraction and could also be important for smooth muscle regulation. We showed earlier that acrylodan-labeled skeletal muscle tropomyosin reports the kinetics of the reversible transitions among the active, intermediate, and inactive states when S1 is rapidly detached from actin tropomyosin. We now show that acrylodan-labeled smooth muscle tropomyosin reports similar transitions among states of actin-tropomyosin. When S1 was rapidly detached from actin-smooth muscle tropomyosin, there was a rapid decrease in acrylodan-tropomyosin fluorescence as the intermediate state became populated. The rate constant for this process was >600 s(-1) at temperatures near 5 degrees C. In the presence of skeletal troponin and EGTA, the decrease in fluorescence was followed by the redevelopment of fluorescence as the inactive state became populated. The apparent rate constant for the fluorescence increase was 14 s(-1) at 5 degrees C. Substituting smooth muscle caldesmon for skeletal muscle troponin produced a similar decrease and re-increase in fluorescence, but the apparent rate constant for the increase was >10 times that observed with troponin. Furthermore, the fluorescence increase was correlated with an increase in the extent of caldesmon attachment as S1-ATP dissociated. Although the measured rate constant appeared to reflect the rate-limiting transition for inactivation, it is unclear if the fluorescence change resulted from caldesmon binding, the movement of tropomyosin over actin, or both. PMID- 21639116 TI - Insecticidal effect and chemical composition of the volatile oil from Bergenia ligulata. AB - The chemical composition of the volatile oil from roots of Bergenia ligulalta was analyzed by GC-MS. A total of 97 compounds were identified. (+)-(6S)-Parasorbic acid (1) (47.45%), isovaleric acid (6.25%), 1,8-cineole (4.24%), (Z)-asarone (3.50%), and terpinen-4-ol (2.96%) were the most prominent constituents. (+)-(6S) Parasorbic acid (1) was isolated and characterized by spectroscopic data. This is the first report of the existence of (+)-(6S)-parasorbic acid in the saxifrage family. The volatile oil and the isolated compound were tested against Drosophila melanogaster . The results obtained showed that the volatile oil from roots could be considered as natural insecticidal effect agents. PMID- 21639117 TI - Cu(I)- and Cu(II)-catalyzed cyclo- and Michael addition reactions of unsaturated beta-ketoesters. AB - The alpha-alkylidene beta-ketoesters 2-carbethoxycyclopentenone (1a) and ethyl 2 benzoylacrylate (1b) react with 1,2-dimethylbutadiene (2) (Diels-Alder), N-benzyl N-(cyclohexylethynyl)-4-methylbenzenesulfonamide (3) (Ficini reaction), ethynyl(phenyl)sulfane (4) ([2 + 2] cycloaddition), and 1,2,5-trimethyl-1H pyrrole (5) (Michael addition) in the presence of copper(I) (6) or copper(II) triflate (7) (1-2 mol %) in dichloromethane. This convenient protocol converts 1a and 1b to the corresponding cycloaddition (8-10) or Michael addition (11) products in good yields after reaction times of 0.5-3 h without requiring purified solvents or inert gas atmosphere. PMID- 21639118 TI - Application of CYP102A1M11H as a tool for the generation of protein adducts of reactive drug metabolites. AB - Covalent binding of reactive metabolites (RMs) to proteins is considered to be one of the important mechanisms by which drugs can cause tissue damage. To facilitate the study of drug-protein adducts, we developed a potentially generic method for producing high levels of covalently modified proteins. A highly active drug metabolizing P450 BM3 mutant (CYP102A1M11H) is used for drug bioactivation. Because of its His-tag, CYP102A1M11H is easily removed by nickel affinity chromatography, facilitating subsequent characterization of the modified target protein. The applicability of our procedure is demonstrated by the trapping of RMs of acetaminophen (APAP), clozapine (CLOZ), and troglitazone (TGZ) with human glutathione-S-transferase P1-1 (hGST P1-1) as the model target protein. Tryptic digests of hGST P1-1 were subjected to analysis by LC-MS/MS and modified peptides identified by the comparative analysis of tryptic peptides of adducted and nonadducted hGST P1-1. Characteristic MS/MS ions of drug-modified peptides were identified by first searching for expected adduct-masses. Unanticipated drug peptide adducts were subsequently identified in an unbiased manner by screening for diagnostic MS/MS ions of modified peptides. Reactive intermediates of APAP and CLOZ adducted to cysteine-47 and mass shifts corresponded to the alkylation of N-acetyl-p-benzoquinone imine (NAPQI) and the CLOZ nitrenium ion, respectively. Adduction of TGZ appeared more complex, yielding three different types of adducts to cysteine-47, two adducts to cysteine-14, and a single adduct to cysteine-101. Together, these findings show that P450 BM3 mutants with high capacity to activate drugs into relevant RMs can be employed to produce protein adducts to study the nucleophilic selectivity of highly reactive electrophiles. PMID- 21639119 TI - Real-space imaging of inelastic Friedel-like surface oscillations emerging from molecular adsorbates. AB - We report real space imaging measurements of inelastic Friedel oscillations. The inelastic electron tunneling spectroscopy, using scanning tunneling microscopy, around dimers of dichlorobenze adsorbates on Au(111) surface display clear spatial modulations that we attribute to inelastic scattering at the molecular sites caused by molecular vibrations. Due to local interactions between the adsorbate and the surface states, the molecular vibrations generate a redistribution of the charge density at energies in a narrow range around the inelastic mode. Our experimental findings are supported by theoretical arguments. PMID- 21639121 TI - Inorganic chemistry and IONiC: an online community bringing cutting-edge research into the classroom. AB - This Viewpoint highlights creative ways that members of the Interactive Online Network of Inorganic Chemists (IONiC) are using journal articles from Inorganic Chemistry to engage undergraduate students in the classroom. We provide information about specific educational materials and networking features available free of charge to the inorganic community on IONiC's web home, the Virtual Inorganic Pedagogical Electronic Resource (VIPEr, www.ionicviper.org ) and describe the benefits of joining this community. PMID- 21639122 TI - Mechanism of resistance to ACCase-inhibiting herbicides in wild oat (Avena fatua ) from Latin America. AB - Whole-plant response of two suspected resistant Avena fatua biotypes from Chile and Mexico to ACCase-inhibiting herbicides [aryloxyphenoxypropionate (APP), cyclohexanedione (CHD), and pinoxaden (PPZ)] and the mechanism behind their resistance were studied. Both dose-response and ACCase enzyme activity assays revealed cross-resistance to the three herbicide families in the biotype from Chile. On the other hand, the wild oat biotype from Mexico exhibited resistance to the APP herbicides and cross-resistance to the CHD herbicides, but no resistance to PPZ. Differences in susceptibility between the two biotypes were unrelated to absorption, translocation, and metabolism of the herbicides. PCR generated fragments of the ACCase CT domain spanning the potential mutations sited in the resistant and susceptible biotypes were sequenced and compared. A point mutation was detected in the aspartic acid triplet at the amino acid position 2078 in the Chilean biotype and in isoleucine at the amino acid position 2041 in the Mexican wild oat biotype, which resulted in a glycine triplet and an asparagine triplet, respectively. On the basis of in vitro assays, the target enzyme (ACCase) in these resistant biotypes contains a herbicide-insensitive form. This is the first reported evidence of resistance to pinoxaden in A. fatua. PMID- 21639123 TI - The use of novel C-methylated spermidine derivatives to investigate the regulation of polyamine metabolism. AB - The polyamines are organic polycations present at millimolar concentrations in eukaryotic cells where they participate in the regulation of vital cellular functions including proliferation and differentiation. Biological evaluation of rationally designed polyamine analogs is one of the cornerstones of polyamine research. Here we have synthesized and characterized novel C-methylated spermidine analogs, that is, 2-methylspermidine, 3-methylspermidine, and 8 methylspermidine. 3-Methylspermidine was found to be metabolically stable in DU145 cells, while 8-methylspermidine was a substrate for spermidine/spermine N(1)-acetyltransferase (SSAT) and 2-methylspermidine was a substrate for both SSAT and acetylpolyamine oxidase. All the analogs induced the splicing of the productive mRNA splice variant of SSAT, overcame growth arrest induced by 72-h treatment with ornithine decarboxylase (ODC) inhibitor alpha difluoromethylornithine, and were transported via the polyamine transporter. Surprisingly, 2-methylspermidine was a weak downregulator of ODC activity in DU145 cells. Our data demonstrates that it is possible to radically alter the biochemical properties of a polyamine analog by changing the position of the methyl group. PMID- 21639124 TI - Tetrasubstituted 2-imidazolones via Ag(I)-catalyzed cycloisomerization of propargylic ureas. AB - A one-pot protocol based on a Ag(I)-catalyzed cycloisomerization of propargylic ureas, derived from secondary propargylamines and isocyanates, was developed for the generation of the 2-imidazolone core. PMID- 21639125 TI - Synthesis of triazafluoranthenones via silver(I)-mediated nonoxidative and oxidative intramolecular palladium-catalyzed cyclizations. AB - Silver(I) fluoride (AgF)-mediated intramolecular nonoxidative and oxidative palladium-catalyzed cyclizations of 1,3-diphenyl- and 8-iodo-1,3 diphenylbenzo[e][1,2,4]triazin-7(1H)-ones 6a (R = H) and 7a (R = I) afford a new 'alkaloid like' ring system 2-phenyl-6H-[1,2,4]triazino[5,6,1-jk]carbazol-6-one 8a (triazafluoranthenone) in 86 and 100% yields, respectively. Furthermore, these cyclization protocols were used to prepare triazafluoranthenone analogues 8b-e bearing dialkylamino, methoxy, and phenylsulfanyl substituents at C-5, which were also independently synthesized from triazafluoranthenone 8a by regioselective nucleophilic addition. Similar AgF-mediated intramolecular nonoxidative and oxidative palladium-catalyzed cyclizations of 8,10-dihydro-1-iodo-10 phenylphenazin-2(7H)-ones 13 gave the new 'alkaloid like' ring system 8H indolo[1,2,3-mn]phenazin-8-one 14 in 80 and 18% yields, respectively. PMID- 21639126 TI - Binuclear Pt-Tl bonded complex with square pyramidal coordination around Pt: a combined multinuclear NMR, EXAFS, UV-Vis, and DFT/TDDFT study in dimethylsulfoxide solution. AB - The structure and bonding of a new Pt-Tl bonded complex formed in dimethylsulfoxide (dmso), (CN)(4)Pt-Tl(dmso)(5)(+), have been studied by multinuclear NMR and UV-vis spectroscopies, and EXAFS measurements in combination with density functional theory (DFT) and time dependent density functional theory (TDDFT) calculations. This complex is formed following the equilibrium reaction Pt(CN)(4)(2-) + Tl(dmso)(6)(3+) ? (CN)(4)Pt-Tl(dmso)(5)(+) + dmso. The stability constant of the Pt-Tl bonded species, as determined using (13)C NMR spectroscopy, amounts to log K = 2.9 +/- 0.2. The (NC)(4)Pt-Tl(dmso)(5)(+) species constitutes the first example of a Pt-Tl bonded cyanide complex in which the sixth coordination position around Pt (in trans with respect to the Tl atom) is not occupied. The spectral parameters confirm the formation of the metal-metal bond, but differ substantially from those measured earlier in aqueous solution for complexes (CN)(5)Pt-Tl(CN)(n)(H(2)O)(x)(n-) (n = 0-3). The (205) Tl NMR chemical shift, delta = 75 ppm, is at extraordinary high field, while spin-spin coupling constant, (1)J(Pt-Tl) = 93 kHz, is the largest measured to date for a Pt-Tl bond in the absence of supporting bridging ligands. The absorption spectrum is dominated by two strong absorption bands in the UV region that are assigned to MMCT (Pt -> Tl) and LMCT (dmso -> Tl) bands, respectively, on the basis of MO and TDDFT calculations. The solution of the complex has a bright yellow color as a result of a shoulder present on the low energy side of the band at 355 nm. The geometry of the (CN)(4)Pt-Tl core can be elucidated from NMR data, but the particular stoichiometry and structure involving the dmso ligands are established by using Tl and Pt L(III)-edge EXAFS measurements. The Pt-Tl bond distance is 2.67(1) A, the Tl-O bond distance is 2.282(6) A, and the Pt-C-N entity is linear with Pt-C and Pt...N distances amounting to 1.969(6) and 3.096(6) A, respectively. Geometry optimizations on the (CN)(4)Pt-Tl(dmso)(5)(+) system by using DFT calculations (B3LYP model) provide bond distances in excellent agreement with the EXAFS data. The four cyanide ligands are located in a square around the Pt atom, while the Tl atom is coordinated in a distorted octahedral fashion with the metal being located 0.40 A above the equatorial plane described by four oxygen atoms of dmso ligands. The four equatorial Tl-O bonds and the four cyano ligands around the Pt atom are arranged in an alternate geometry. The coordination environment around Pt may be considered as being square pyramidal, where the apical position is occupied by the Tl atom. The optimized geometry of (CN)(4)Pt-Tl(dmso)(5)(+) is asymmetrical (C(1) point group). This low symmetry might be responsible for the unusually large NMR linewidths observed due to intramolecular chemical exchange processes. The nature of the Pt-Tl bond has been studied by MO analysis. The metal-metal bond formation in (CN)(4)Pt Tl(dmso)(5)(+) can be simply interpreted as the result of a Pt(5d(z(2)))(2) -> Tl(6s)(0) donation. This bonding scheme may rationalize the smaller thermodynamic stability of this adduct compared to the related complexes with (CN)(5)Pt-Tl entity, where the linear C-Pt-Tl unit constitutes a very stable bonding system. PMID- 21639127 TI - Rectangular chain packing of methyl-branched paraffins: persistence of an interchain interaction and forms of disorder. AB - The rectangular crystal packing of methyl-branched paraffins in their orthorhombic forms is studied systematically by electron diffraction to show that, irrespective of branch position on the chain, a close interaction of chain double rows occurs, placing the branch in the space between two chain ends. If the chain branches occur near the ends, the structure can slowly rearrange into a true bilayer. If the branch occurs near the center, then there are a large number of intermediate "nematocrystalline" disordered forms that are possible before the final ordered layered packing. PMID- 21639128 TI - Using chemical shifts to determine structural changes in proteins upon complex formation. AB - Methods for determining protein structures using only chemical shift information are progressively becoming more accurate and reliable. A major problem, however, in the use of chemical shifts for the determination of the structures of protein complexes is that the changes in the chemical shifts upon binding tend to be rather limited and indeed often smaller than the standard errors made in the predictions of chemical shifts corresponding to given structures. We present a procedure that, despite this problem, enables one to use of chemical shifts to determine accurately the conformational changes that take place upon complex formation. PMID- 21639129 TI - Phosphorescent iridium(III) complex with an N^O ligand as a Hg(2+)-selective chemodosimeter and logic gate. AB - Phosphorescent iridium(III) complexes have been attracting increasing attention in applications as luminescent chemosensors. However, no instance of an iridium(III) complex being used as a molecular logic gate has hitherto been reported. In the present study, two iridium(III) complexes, [Ir(ppy)(2)(PBT)] and [Ir(ppy)(2)(PBO)], have been synthesized (PBT, 2-(2-Hydroxyphenyl)-benzothiazole; PBO, 2-(2-hydroxyphenyl)-benzoxazole), and their chemical structures have been characterized by single-crystal X-ray analysis. Theoretical calculations and detailed studies of the photophysical and electrochemical properties of these two complexes have shown that the N^O ligands dominate their luminescence emission properties. Moreover, [Ir(ppy)(2)(PBT)], containing a sulfur atom in the N^O ligand, can serve as a highly selective chemodosimeter for Hg(2+) with ratiometric and naked-eye detection, which is associated with the dissociation of the N^O ligand PBT from the complex. Furthermore, complex [Ir(ppy)(2)(PBT)] has been further developed as an AND and INHIBIT logic gate with Hg(2+) and histidine as inputs. PMID- 21639130 TI - Dual responsive nanostructured surfaces for biomedical applications. AB - In this paper, we describe the construction and characteristics of thermoresponsive, thin nanostructured films prepared by layer-by-layer sequential assembly of chitosan-graft-NIPAAm and alginate. FTIR and (1)H NMR spectra have confirmed the introduction of NIPAAm moieties onto the chitosan backbone. The LCST of the synthesized copolymer was found to be around 31-33 degrees C. The formation of the polyelectrolyte multilayers containing the copolymer and alginate was followed in situ by quartz crystal microbalance with dissipation monitoring technique and ex situ by UV-vis measurements. Our results revealed the linear increase of the multilayer film growth and the influence of the presence of salt. Moreover, AFM analysis has confirmed that PNIPAAm is able to reconform upon temperature swaps even when combined with other layers in a polyelectrolyte multilayer, demonstrating that the nanoassemblies are thermoresponsive. Preliminary results showed that, upon reducing culture temperature below PNIPAAm LCST, a gradual detachment of cell sheets from these PNIPAAm-based coatings has occurred. PMID- 21639131 TI - Identification of a naturally occurring quinazolin-4(3H)-one firefly luciferase inhibitor. AB - A prefractionated Streptomyces-derived extract was initially identified as being active using a luciferase-based AMP-activated protein kinase (AMPK) assay. Bioassay-guided fractionation led to the isolation of the new compound quinazolin 4(3H)-one (1) as the active component. However, 1 was shown to have potent firefly luciferase inhibitory activity with no effect on AMPK. This is the first report of a natural luciferase inhibitor. PMID- 21639132 TI - Detection of single nucleotide polymorphism using tension-dependent stochastic behavior of a single-molecule template. AB - Single nucleotide polymorphism (SNP) is the most common genetic variation among individuals. The association of SNP with individual's response to pathogens, phenotypic variations, and gene functions emphasizes the importance of sensitive and reliable SNP detection for biomedical diagnosis and therapy. To increase sensitivity, most approaches employ amplification steps, such as PCR, to generate detectable signals that are usually ensemble-averaged. Introduction of amplification steps increases the complexity of a system, whereas ensemble averaging of signals often suffers from background interference. Here, we have exploited the stochastic behavior of a single-molecule probe to recognize SNP sequence in a microfluidic platform using a laser-tweezers instrument. The detection relies on on-off mechanical signals that provide little background interference and high specificity between wild type and SNP sequences. The microfluidic setting allows multiplex sensing and in situ recycling of the SNP probe. As a proof-of-concept, we have detected as low as 100 pM of an SNP target associated with coronary heart diseases within half an hour without any amplification steps. The mechanical signal permits the detection of single mutations involving either G/C or A/T pairs. We anticipate this system has the capacity to function as a highly sensitive generic biosensor after incorporation of a specific recognition element, such as an aptamer for example. PMID- 21639133 TI - Carbon nanotubes activate store-operated calcium entry in human blood platelets. AB - Carbon nanotubes (CNTs) are known to potentiate arterial thrombosis in animal models, which raises serious safety issues concerning environmental or occupational exposure to CNTs and their use in various biomedical applications. We have shown previously that different CNTs, but not fullerene (nC60), induce the aggregation of human blood platelets. To date, however, a mechanism of potentially thrombogenic CNT-induced platelet activation has not been elucidated. Here we show that pristine multiwalled CNTs (MWCNTs) penetrate platelet plasma membrane without any discernible damage but interact with the dense tubular system (DTS) causing depletion of platelet intracellular Ca(2+) stores. This process is accompanied by the clustering of stromal interaction molecule 1 (STIM1) colocalized with Orai1, indicating the activation of store-operated Ca(2+) entry (SOCE). Our findings reveal the molecular mechanism of CNT-induced platelet activation which is critical in the evaluation of the biocompatibility of carbon nanomaterials with blood. PMID- 21639135 TI - Iridium complexes of the doubly cyclometalated NHC ligand IMes''. AB - Two consecutive C-H bond activations at the coordination sphere of Ir transform the commonly employed NHC ligand IMes into the new kappa(3)-C fac-coordinating ligand IMes''. The preparation and structure of Ir(III) complexes featuring this ligand together with selected reactions toward small molecules that illustrate their reactivity keys are described. PMID- 21639134 TI - Discovery and optimization of sulfonyl acrylonitriles as selective, covalent inhibitors of protein phosphatase methylesterase-1. AB - The serine hydrolase protein phosphatase methylesterase-1 (PME-1) regulates the methylesterification state of protein phosphatase 2A (PP2A) and has been implicated in cancer and Alzheimer's disease. We recently reported a fluorescence polarization-activity-based protein profiling (fluopol-ABPP) high-throughput screen for PME-1 that uncovered a remarkably potent and selective class of aza beta-lactam (ABL) PME-1 inhibitors. Here, we describe a distinct set of sulfonyl acrylonitrile inhibitors that also emerged from this screen. The optimized compound, 28 (AMZ30), selectively inactivates PME-1 and reduces the demethylated form of PP2A in living cells. Considering that 28 is structurally unrelated to ABL inhibitors of PME-1, these agents, together, provide a valuable set of pharmacological probes to study the role of methylation in regulating PP2A function. We furthermore observed that several serine hydrolases were sensitive to analogues of 28, suggesting that more extensive structural exploration of the sulfonyl acrylonitrile chemotype may result in useful inhibitors for other members of this large enzyme class. PMID- 21639136 TI - Modeling DNA-bending in the nucleosome: role of AA periodicity. AB - This paper uses atomistic molecular mechanics within the framework of the JUMNA model to study the bending properties of DNA segments, with emphasis on understanding the role of the 10 bp periodicity associated with AA repeats that has been found to dominate in nucleosomal DNA. The calculations impose a bending potential on 18 bp segments that is consistent with nucleosome structures (i.e., radius of curvature of 4.1 nm), and then determine the energies of the minimum energy structures for different values of the rotational register (a measure of the direction of bending of the DNA) subject to forces derived from the Amber force field (parm99bsc0). The results show that sequences that contain the 10 bp repeats but are otherwise random have a narrow distribution of rotational register values that minimize the energy such that it is possible to combine several minimized structures to give the 147 bp nearly planar loop structure of the nucleosome. The rotational register values that lead to minimum bending energy with 10 bp AA repeats have a narrower minor groove, which points toward the histone interior at the positions of the AA repeats, which is a result that matches the experiments. The calculations also show that these sequences have a relatively flat potential energy landscape for bending to a 4.1 nm radius of curvature. Random sequences that do not have the 10 bp AA repeats have less stable bent structures, and a flat rotational register distribution, such that low energy nearly planar loops are less likely. PMID- 21639137 TI - A glimpse at the chemistry of GeH2 in solution. Direct detection of an intramolecular germylene-alkene pi-complex. AB - The photochemistry of 3-methyl-4-phenyl-1-germacyclopent-3-ene (4) and a deuterium-labeled derivative (4-d(2)) has been studied in solution by steady state and laser flash photolysis methods, with the goal of detecting the parent germylene (GeH(2)) directly and studying its reactivity in solution. Photolysis of 4 in C(6)D(12) containing acetic acid (AcOH) or methanol (MeOH) affords 2 methyl-3-phenyl-1,3-butadiene (6) and the O-H insertion products ROGeH(3) (R = Me or Ac) in yields of ca. 60% and 15-30%, respectively, along with numerous minor products which the deuterium-labeling studies suggest are mainly derived from hydrogermylation processes involving GeH(2) and diene 6. The reaction with AcOH also affords H(2) in ca. 20% yield, while HD is obtained from 4-d(2) under similar conditions. Photolysis of 4 in THF-d(8) containing AcOH affords AcOGeH(3) and 6 exclusively, indicating that the nucleophilic solvent assists the extrusion of GeH(2) from 4 and alters the mechanism of the trapping reaction with AcOH compared to that in cyclohexane. Laser flash photolysis of 4 in hexanes yields a promptly formed transient exhibiting lambda(max) ~ 460 nm, which decays on the microsecond time scale with the concomitant growth of a second, much longer-lived transient exhibiting lambda(max) ~ 390 nm. The spectrum and reactivity of the 460 nm species toward various germylene trapping agents are inconsistent with those expected for free GeH(2); rather, the transient is assigned to an intramolecular Ge(II)-alkene pi-complex of one of the isomeric substituted hydridogermylenes derived from a solvent-cage reaction between GeH(2) and its diene (6) coproduct, formed by addition of HGe-H across one of the C=C bonds. These conclusions are supported by the results of DFT calculations of the thermochemistry associated with pi-complexation of GeH(2) with 6 and the formation of the isomeric vinylgermiranes and 1,2-hydrogermylation products. A different species is observed upon laser photolysis of 4 in THF solution and is assigned to the GeH(2) THF complex on the basis of its UV-vis spectrum and rate constants for its reaction with AcOH and AcOD. PMID- 21639138 TI - Thermoresponsive dynamers: thermally induced, reversible chain elongation of amphiphilic poly(acylhydrazones). AB - A nanostructured poly(acylhydrazone), which is reversibly formed in acidic aqueous solution from di(aldehyde) and di(acylhydrazine) monomers with appended hexaglyme groups, was found to display lower critical solution (LCS) behavior. Remarkably, under acidic conditions in which polymerization is reversible, large and reversible molecular weight (M(w)) increases were observed in response to elevated temperatures, both below and above the LCS temperature. No variation in M(w) was evident under neutral and alkaline conditions, in which the acylhydrazone condensation is essentially irreversible. Results of turbidometry studies, size-exclusion chromatography-multiangle laser light scattering (SEC MALLS), and transmission electron microscopy (TEM) suggest that heating the polymer below the LCS temperature leads to polymer growth with preservation of the characteristic nanostructured morphology, whereas the onset of the microphase separated state causes a fundamental change in morphology, in which the polymer chains aggregate into larger bundles and fibers. van't Hoff analysis of a small molecule model system indicates that the acylhydrazone condensation is enthalpy driven (DeltaH(eq) = -8.2 +/- 0.2 kcal.mol(-1) and DeltaS(eq) = -11.1 +/- 0.4 = cal.mol(-1).K(-1)), which suggests that the observed polymer growth with temperature is not a consequence of the intrinsic thermodynamics of the intermonomer linkage but is likely the result of the thermoresponsive characteristics conferred by the multiple hexaglyme groups. The system described displays double control of the dynamer state by two orthogonal agents, heat and protons (pH). It also represents a prototype for dynamic materials displaying multiple control adaptive behavior. PMID- 21639139 TI - Cell-specific targeting by heterobivalent ligands. AB - Current cancer therapies exploit either differential metabolism or targeting to specific individual gene products that are overexpressed in aberrant cells. The work described herein proposes an alternative approach--to specifically target combinations of cell-surface receptors using heteromultivalent ligands ("receptor combination approach"). As a proof-of-concept that functionally unrelated receptors can be noncovalently cross-linked with high avidity and specificity, a series of heterobivalent ligands (htBVLs) were constructed from analogues of the melanocortin peptide ligand ([Nle(4), dPhe(7)]-alpha-MSH) and the cholecystokinin peptide ligand (CCK-8). Binding of these ligands to cells expressing the human Melanocortin-4 receptor and the Cholecystokinin-2 receptor was analyzed. The MSH(7) and CCK(6) were tethered with linkers of varying rigidity and length, constructed from natural and/or synthetic building blocks. Modeling data suggest that a linker length of 20-50 A is needed to simultaneously bind these two different G-protein coupled receptors (GPCRs). These ligands exhibited up to 24 fold enhancement in binding affinity to cells that expressed both (bivalent binding), compared to cells with only one (monovalent binding) of the cognate receptors. The htBVLs had up to 50-fold higher affinity than that of a monomeric CCK ligand, i.e., Ac-CCK(6)-NH(2). Cell-surface targeting of these two cell types with labeled heteromultivalent ligand demonstrated high avidity and specificity, thereby validating the receptor combination approach. This ability to noncovalently cross-link heterologous receptors and target individual cells using a receptor combination approach opens up new possibilities for specific cell targeting in vivo for therapy or imaging. PMID- 21639140 TI - Soluble and bound phenolic compounds in different Bolivian purple corn ( Zea mays L.) cultivars. AB - In nine Bolivian purple corn ( Zea mays L.) varieties the content of phenolic compounds as well as the anthocyanin composition has been determined. The phenotypes under investigation included four red and five blue varieties (Kulli, Ayzuma, Paru, Tuimuru, Oke, Huaca Songo, Colorado, Huillcaparu, and Checchi). In purple corn, phenolic compounds were highly concentrated in cell walls. Thus, simultaneous determination of soluble and bound-form phenolics is essential for analysis, extraction, and quantification. The present study reports the determination of soluble and insoluble-bound fraction of phenolic compounds by HPLC-DAD and HPLC-ESI-MS(n) in Bolivian purple corn varieties. Enzymatic, thermal, and alkaline hydrolyses were used to obtain the cell wall-linked phenolic compounds. Ferulic acid values ranged from 132.9 to 298.4 mg/100 g, and p-coumaric acid contents varied between 251.8 and 607.5 mg/100 g dry weight (DW), respectively, and were identified as the main nonanthocyanin phenolics. The total content of phenolic compounds ranged from 311.0 to 817.6 mg gallic acid equivalents (GAE)/100 g DW, and the percentage contribution of bound to total phenolics varied from 62.1 to 86.6%. The total monomeric anthocyanin content ranged from 1.9 to 71.7 mg cyanidin-3-glucoside equivalents/100 g DW. Anthocyanin profiles are almost the same among the different samples. Differences are observed only in the relative percentage of each anthocyanin. Cyanidin-3 glucoside and its malonated derivative were detected as major anthocyanins. Several dimalonylated monoglucosides of cyanidin, peonidin, and pelargonidin were present as minor constituents. PMID- 21639141 TI - A normal mode-based geometric simulation approach for exploring biologically relevant conformational transitions in proteins. AB - A three-step approach for multiscale modeling of protein conformational changes is presented that incorporates information about preferred directions of protein motions into a geometric simulation algorithm. The first two steps are based on a rigid cluster normal-mode analysis (RCNMA). Low-frequency normal modes are used in the third step (NMSim) to extend the recently introduced idea of constrained geometric simulations of diffusive motions in proteins by biasing backbone motions of the protein, whereas side-chain motions are biased toward favorable rotamer states. The generated structures are iteratively corrected regarding steric clashes and stereochemical constraint violations. The approach allows performing three simulation types: unbiased exploration of conformational space; pathway generation by a targeted simulation; and radius of gyration-guided simulation. When applied to a data set of proteins with experimentally observed conformational changes, conformational variabilities are reproduced very well for 4 out of 5 proteins that show domain motions, with correlation coefficients r > 0.70 and as high as r = 0.92 in the case of adenylate kinase. In 7 out of 8 cases, NMSim simulations starting from unbound structures are able to sample conformations that are similar (root-mean-square deviation = 1.0-3.1 A) to ligand bound conformations. An NMSim generated pathway of conformational change of adenylate kinase correctly describes the sequence of domain closing. The NMSim approach is a computationally efficient alternative to molecular dynamics simulations for conformational sampling of proteins. The generated conformations and pathways of conformational transitions can serve as input to docking approaches or as starting points for more sophisticated sampling techniques. PMID- 21639142 TI - Characterizing the dynamics and ligand-specific interactions in the human leukocyte elastase through molecular dynamics simulations. AB - The human leukocyte elastase (HLE), a neutrophil serine protease of the chymotrypsin superfamily, is a major therapeutic target for a number of inflammatory diseases, such as chronic obstructive pulmonary disease (COPD). In this work, we present a comparative explicit water molecular dynamics (MD) study on the free and inhibitor-bound HLE. Knowledge of the flexibility and conformational changes induced by this irreversible inhibitor, whether in a prebound stage or covalently bound at the enzyme binding site, encases fundamental biological interest and is particularly relevant to ongoing structure based drug design studies. Our results suggest that HLE operates by an induced fit mechanism with direct intervention of a surface loop which is open toward the solvent in the free enzyme and closed while in the presence of the ligand. MM PBSA free energy calculations furthermore elucidate the energetic contributions to the distinct conformations adopted by this loop. Additionally, a survey of the major contributions to the inhibitor binding free energies was attained. Our findings enforce the need to account for HLE flexibility, whether through the use of MD-generated ensembles of HLE conformations as targets for molecular docking or via sophisticated flexible-docking algorithms. We anticipate that inclusion of the observed HLE dynamic behavior into future drug design methodologies will have a relevant impact in the development of novel, more efficient, inhibitors. PMID- 21639143 TI - Subtraction of DC conductivity and annealing: approaches to identify Johari Goldstein relaxation in amorphous trehalose. AB - Amorphous trehalose finds extensive use as a stabilizer of biomolecules including proteins and phospholipids. Hypothesizing that molecular mobility is a determinant of its stability, dynamic dielectric spectroscopy (DDS) was used to map the different modes of mobility. Isothermal dielectric relaxation profiles of amorphous trehalose were obtained, over the frequency range of 10(-1)-10(7) Hz, and at temperatures ranging from 30-170 degrees C. At temperatures close to the glass transition (T(g)), the alpha-relaxation was not readily discernible due to interference from dc conductivity. We used Kramers-Kronig transformation that enabled not only the complete resolution of alpha-relaxation but also the identification of an excess wing, in the high frequency tail of alpha-relaxation. On annealing, this excess wing developed into a partially resolved and hitherto unidentified beta-relaxation peak. This peak, due to its position in the dielectric spectrum, its annealing time dependence and the good agreement with the calculated independent relaxation time, was assigned to the Johari-Goldstein process. This work demonstrates the utility of conductivity subtraction coupled with sub-T(g) annealing to successfully study all the modes of mobility in amorphous trehalose. This approach can potentially be extended to situations wherein dc conductivity impedes the complete characterization of molecular mobility. PMID- 21639144 TI - Synthesis, in vitro evaluation, and in vivo metabolism of fluor/quencher compounds containing IRDye 800CW and Black Hole Quencher-3 (BHQ-3). AB - Protease-cleavable peptides containing a suitable fluor/quencher (Fl/Q) pair are optically dark until cleaved by their target protease, generating fluorescence. This approach has been used with many Fl/Q pairs, but little has been reported with IRDye 800CW, a popular near-infrared (NIR) fluor. We explored the use of the azo-bond-containing Black Hole Quencher 3 (BHQ-3) as a quencher for IRDye 800CW and found that IRDye 800CW/BHQ-3 is a suitable Fl/Q pair, despite the lack of proper spectral overlap for fluorescence resonance energy transfer (FRET) applications. Cleavage of IRDye 800CW-PLGLK(BHQ-3)AR-NH(2) (8) and its D-arginine (Darg) analogue (9) by matrix metalloproteinases (MMPs) in vitro yielded the expected cleavage fragments. In vivo, extensive metabolism was found. Significant decomposition of a "non-cleavable" control IRDye 800CW-(1,13-diamino-4,7,10 trioxatridecane)-BHQ-3 (10) was evident in plasma of normal mice by 3 min post injection. The major metabolite showed a m/z and UV/vis spectrum consistent with azo bond cleavage in the BHQ-3 moiety. Preparation of an authentic standard of this metabolite (11) confirmed the assignment. Although the IRDye 800CW/BHQ-3 constructs showed efficient contact quenching prior to enzymatic cleavage, BHQ-3 should be used with caution in vivo, due to instability of its azo bond. PMID- 21639145 TI - Development of an enhanced chemiluminescence ELISA for the rapid detection of acrylamide in food products. AB - In this work, a polyclonal antibody for acrylamide (AA) was obtained by immunization of rabbits with N-acryloxysuccinimide (NAS) and keyhole limpet hemocyanin (KLH) conjugate. A direct enzyme-linked immunosorbent assay (ELISA) based on this antibody was developed with enhanced chemiluminescent (ECL) detection of AA in food samples. Assay conditions, such as concentrations of antibody and enzyme conjugate and competition time, were optimized. The effects of ionic strength and pH value were investigated. The optimized ECL-ELISA system allowed AA determination in a linear working range of 26.3-221.1 ng mL(-1) with an IC(50) value of 60.6 ng mL(-1) and a limit of detection of 18.6 ng mL(-1). Good recoveries with spiked food samples were obtained with a recovery range from 74.4 to 98.1%, and these results correlated well with those obtained using an HPLC method. This indicates that ECL-ELISA is applicable to the specific detection and routine monitoring of AA in food samples. PMID- 21639146 TI - Oxidation and loss of heme in soluble guanylyl cyclase from Manduca sexta. AB - Oxidation and loss of heme in soluble guanylyl/guanylate cyclase (sGC), the nitric oxide receptor, is thought to be a major contributor to cardiovascular disease and is the target of compounds BAY 58-2667 and HMR1766. Using spectroelectrochemical titration, we found a truncated sGC to be highly stable in the ferrous state (234 mV) and to bind ferrous heme tightly even in the presence of NO, despite the NO-induced release of the proximal histidine. In contrast, oxidized sGC readily loses ferric heme to myoglobin (0.47 +/- 0.02 h(-1)). Peroxynitrite, the presumed cellular oxidant, readily oxidizes sGC in 5 mM glutathione. PMID- 21639147 TI - Entrapment of a water wire in a hydrophobic peptide channel with an aromatic lining. AB - A one-dimensional water wire has been characterized by X-ray diffraction in single crystals of the tripeptide Ac-Phe-Pro-Trp-OMe. Crystals in the hexagonal space group P6(5) reveal a central hydrophobic channel lined by aromatic residues which entraps an approximately linear array of hydrogen bonded water molecules. The absence of any significant van der Waals contact with the channel walls suggests that the dominant interaction between the "water wire" and "peptide nanotube" is electrostatic in origin. An energy difference of 16 kJ mol(-1) is estimated for the distinct orientations of the water wire dipole with respect to the macrodipole of the peptide nanotube. The structural model suggests that Grotthuss type proton conduction may, through constricted hydrophobic channels, be facilitated by concerted, rotational reorientation of water molecules. PMID- 21639148 TI - Accumulation and distribution pattern of macro- and microelements and trace elements in Vitis vinifera L. cv. Chardonnay berries. AB - This paper describes the accumulation pattern of 42 mineral elements in Vitis vinifera L. berries during development and ripening and their distribution in berry skin, seeds, and flesh around harvest time. Grape berries were sampled in two different vineyards with alkaline soil and analyzed using a ICP-MS. Although elemental amounts were significantly different in the grapes from the two vineyards, the accumulation pattern and percentage distribution in different parts of the berries were generally quite similar. Ba, Eu, Sr, Ca, Mg, Mn, and Zn accumulate prior to veraison. Al, Ce, Dy, Er, Ga, Gd, Ho, La, Nd, Pr, Sm, Sn, Zr, Th, Tm, U, Y, and Yb accumulate mainly prior to veraison but also during ripening. Ag, As, B, Cd, Cs, Cu, Fe, Ge, Hg, K, Li, Na, P, Rb, Sb, Se, and Tl accumulate progressively during growth and ripening. With regard to distribution, Ba, Ca, Eu, Fe, Mn, P, Sr, and Zn accumulate mainly in the seeds, Al, B, Ga, Sn, and the rare earths analyzed, except for Eu, accumulate mainly in the skin, and Ag, As, Cd, Cs, Cu, Ge, Hg, K, Li, Mg, Na, Rb, Sb, Se, Th, Tl, U, and Zr accumulate mainly in the flesh. A joint representation of the accumulation and distribution patterns for the elements in the berry is also given. PMID- 21639149 TI - Nonnative interactions in the FF domain folding pathway from an atomic resolution structure of a sparsely populated intermediate: an NMR relaxation dispersion study. AB - Several all-helical single-domain proteins have been shown to fold rapidly (microsecond time scale) to a compact intermediate state and subsequently rearrange more slowly to the native conformation. An understanding of this process has been hindered by difficulties in experimental studies of intermediates in cases where they are both low-populated and only transiently formed. One such example is provided by the on-pathway folding intermediate of the small four-helix bundle FF domain from HYPA/FBP11 that is populated at several percent with a millisecond lifetime at room temperature. Here we have studied the L24A mutant that has been shown previously to form nonnative interactions in the folding transition state. A suite of Carr-Purcell-Meiboom Gill relaxation dispersion NMR experiments have been used to measure backbone chemical shifts and amide bond vector orientations of the invisible folding intermediate that form the input restraints in calculations of atomic resolution models of its structure. Despite the fact that the intermediate structure has many features that are similar to that of the native state, a set of nonnative contacts is observed that is even more extensive than noted previously for the wild-type (WT) folding intermediate. Such nonnative interactions, which must be broken prior to adoption of the native conformation, explain why the transition from the intermediate state to the native conformer (millisecond time scale) is significantly slower than from the unfolded ensemble to the intermediate and why the L24A mutant folds more slowly than the WT. PMID- 21639151 TI - Consecutive infrared multiphoton dissociations in a fourier transform ion cyclotron resonance mass spectrometer. AB - Consecutive infrared multiphoton dissociations (IRMPD) may be observed in a Fourier transform ion cyclotron resonance mass spectrometer (FTICR). This is the IRMPD equivalent of previous MS(n)() experiments using CID. This work presents a versatile technique, using a bistable shutter to gate ON and OFF a continuous wave (CW) CO(2) laser for multiple irradiation periods of 0.1-1000 s duration. Consecutive photodissociations, up to MS(4), are demonstrated for the proton bound dimer of diethyl ether and the resulting fragment ions. The photoproducts are formed close to the center of the FTICR cell, resulting in high product ion recovery efficiency. This differs from CID products, which are formed throughout the FTICR cell causing reisolation/detection problems. The fragmentation resulting from the use of low-intensity, CW, infrared laser radiation is shown to be much more energy selective than CID. Photodissociation of C(2)H(5)OH(2)(+) ion produces the lowest energy product ion exclusively, even though the two product channels differ only by ~5 kcal/mol. Low-energy CID, however, produces a mixture of C(2)H(5)(+) and H(3)O(+) products in the ratio of 1.3:1. Hence, the higher energy pathway (C(2)H(5)(+)) is substantially favored. The current results indicate that this IRMPD MS(n)() technique may be successfully applied to large biomolecules prepared by electrospray or MALDI. PMID- 21639150 TI - Biomimetic hydrogels with immobilized ephrinA1 for therapeutic angiogenesis. AB - The formation of a microvasculature is regulated in large part by cell-cell interactions. Ephrins and their Eph receptors mediate cell adhesion, repulsion, and migration, all critical processes in angiogenesis. (1) Here we use a covalently immobilized ephrinA1, conjugated to poly(ethylene glycol), to induce vessel formation both in vitro and in vivo in poly(ethylene glycol) diacrylate (PEGDA) hydrogels. Human umbilical vein endothelial cell (HUVEC) tubulogenesis in matrix metalloproteinase-sensitive hydrogels was visualized from 6 h to 7 days in response to three different concentrations of PEG-ephrinA1. The deposition of extracellular matrix proteins collagen IV and laminin that stabilize tubule formation were imaged, quantified, and found to be dependent on PEG-ephrinA1 concentration. To confirm the importance of the EphA2-ephrinA1 interaction in tubule formation, soluble EphA2 was used to disrupt the EphA2-ephrinA1 interaction between a coculture of HUVEC and human brain vascular pericyte cells. HUVECs seeded onto PEGDA hydrogels displayed a dose-dependent reduction in tubule formation in response to the soluble EphA2. Finally, hydrogels with releasable platelet-derived growth factor (PDGF), immobilized RGDS, and covalently immobilized PEG-ephrinA1 were implanted into the mouse cornea micropocket. These hydrogels induced a more robust vascular response with an increase in vessel density as compared with hydrogels with releasable PDGF alone. As such, PEG ephrinA1 may represent a promising molecule to regulate cell adhesion and migration for formation of a microvasculature in tissue-engineered constructs. PMID- 21639152 TI - Elemental speciation analysis by multicapillary gas chromatography with microwave induced plasma atomic spectrometric detection. AB - Multicapillary column gas chromatography (MC-GC)/microwave-induced plasma atomic emission spectrometry (MIP AES) was developed for fast speciation analysis of organotin compounds in the environment. Ethylated butyltin compounds could be separated isothermally within less than 30 s (instead of ~5-10 min) without sacrificing either the resolution or the sample capacity of conventional capillary GC with oven temperature gradient programming. Careful optimization of the pressure and temperature GC program allowed a comprehensive organotin speciation analysis including phenyltin compounds within less than 2.5 min, increasing the sample throughput 6-fold. Compatibility of MC-GC with an MIP atomic emission detector (MIP-AED) was discussed. MC-GC/MIP-AES was validated for the analysis of sediment (PACS-1 and BCR 462) and biological (NIES11) certified reference materials. PMID- 21639153 TI - Ion-pair extraction system for the mutual separation of lanthanides using divalent quadridentate schiff bases. AB - For the selective extraction of trivalent lanthanides, the use of quadridentate divalent phenolic Schiff bases, such as N,N'-bis(5 nitrosalicilidene)ethylenediamine (H(2)Nsalen) and N,N'-bis(5-nitrosalicilidene) o-phenylenediamine (H(2)Nsaloph), was investigated. Lanthanides made anionic 1:2 complexes with these ligands and could be extracted into nitrobenzene as ion pairs with a suitable monovalent countercation in the aqueous phase, after which the ion-pair was dissociated almost perfectly. Although the order of the extractability of lanthanides between these ligands was H(2)Nsaloph > H(2)Nsalen, the mutual selectivity of them in the H(2)Nsalen system was higher than that in H(2)Nsaloph. Furthermore, with decreasing the size of the countercation, the mutual selectivity was enhanced although the extractability was lowered. A H(2)Nsalen-KCl extraction system, selected as an example, showed selectivity between lanthanides comparable with those of the better of other systems reported previously. PMID- 21639154 TI - Spectroelectrochemical sensing based on multimode selectivity simultaneously achievable in a single device. 2. Demonstration of selectivity in the presence of direct interferences. AB - Three modes of selectivity based on charge-selective partitioning, electrolysis potential, and spectral absorption wavelength were demonstrated simultaneously in a new type of spectroelectrochemical sensor. Operation and performance of the three modes of selectivity for detection of analytes in the presence of direct interferences were investigated using binary mixture systems. These binary mixtures consisted of Fe(CN)(6)(3-) and Ru(bpy)(3)(2+) and of Fe(CN)(6)(4-) and Ru(CN)(6)(4)(-) in aqueous solutions. Results on the Fe(CN)(6)(3-)/Ru(bpy)(3)(2+) binary mixture showed that an anion-exchange coating consisting of PDMDAAC-SiO(2) [where PDMDAAC is poly(dimethyldiallylammonium chloride)] and a cation-exchange coating consisting of Nafion-SiO(2) can trap and preconcentrate analytes with charge selection. At the same time, such coatings exclude interferences carrying the same type of charge as that of the exchange sites in the sensor coating. Using the Fe(CN)(6)(4-)/Ru(CN)(6)(4-) binary mixture, the Fe(CN)(6)(4-) component can be selectively detected by restricting the modulation potential cycled to a range specific to the redox-active Fe(CN)(6)(4-) component and simultaneously monitoring the optical response at the overlapping wavelength of 420 nm. It was also shown that, when the wavelength for optical monitoring was chosen as 500 nm, which is specific to the Ru(CN)(6)(4-) component, interference from the Fe(CN)(6)(4-) component for spectroelectrochemical detection of Ru(CN)(6)(4-) was significantly suppressed, even though the cyclic modulation potential encompassed the redox range for the Fe(CN)(6)(4-) component. PMID- 21639155 TI - Electrochemical quartz crystal impedance study of redox hydrogel mediators for amperometric enzyme electrodes. AB - Quartz crystal impedance around the resonant frequency at 10 MHz of a composite quartz crystal resonator has been studied simultaneously with cyclic voltammetry. A modified quartz crystal with a redox hydrogel (poly(allylamine)-ferrocene cross linked with glucose oxidase) and immersed in liquid electrolyte was used. Impedance parameters (R(f) and X(L)((f))) of the surface redox gel film were obtained by fitting the resonator transfer function |V(o)/V(i)| vs omega to a BVD equivalent circuit and analyzed with the multiple nonpiezoelectric layer model of Martin. Two limiting hydrogel layers of the same composition were studied while oxidizing and reducing the ferrocene/ferricenium moieties attached to the swollen polymer backbone: thin and thick redox hydrogel films. For the thin films, the Sauerbrey approximation was valid. The mass/thickness and film viscosity changes that resulted from the anion and water exchange were evaluated while redox switching the polymer on the assumption of negligible storage modulus G' and a density of 1. For thick gel layers, on the other hand, the penetration depth of the acoustic wave was far less than the film thickness, and a liquid-like behavior was apparent. Film storage modulus and film loss modulus were simultaneously evaluated with the cyclic voltammetry. PMID- 21639156 TI - Separation and Pulsed Amperometric Detection of Alditols and Carbohydrates by Anion-Exchange Chromatography Using Alkaline Mobile Phases Modified with Ba(II), Sr(II), and Ca(II) Ions. AB - The effect of some divalent nonelectroactive cations (DNCs) in the anion-exchange chromatographic separations with alkaline mobile phases of carbohydrates and alditols was investigated; the ions examined at a typical concentration of 1 mM were Ca(II), Ba(II), and Sr(II). The use of these cations in the eluent as their corresponding acetates or nitrates has been found to yield at least a two-fold effect. First, the peak symmetry and concurrently the column efficiency is greatly improved. Second, the presence of Ba(II) or Sr(II) significantly enhances the response of all analytes investigated in pulsed amperometry at a gold working electrode. The action of the DNCs on the separation seems to be related to a very effective removal of carbonate ion from the alkaline eluent and, especially, their ability to complex cyclic and acyclic polyhydroxy compounds. Efficiency estimated from data calculated at 10% of peak height can increase by as much as 25% with a comparably lower RSD (15%, n = 9). From the viewpoint of the separation efficiency and reproducibility of chromatographic data, all divalent inorganic ions employed were well-behaved. However, only the use of Ba(II) or Sr(II) is recommended since alkaline mobile phases containing Ca(II) ion negatively affect the gold electrode response of some analytes. PMID- 21639157 TI - Effect of Ca(II), Sr(II), and Ba(II) on the Pulsed Amperometric Detection of Alditols and Carbohydrates at a Gold Electrode in Alkaline Solutions. AB - The effects of some divalent nonelectroactive cations (DNCs), such as Sr(II), Ba(II), and Ca(II) on the electrochemical oxidation of alditols and carbohydrates at gold electrodes in pulsed amperometric detection have been investigated. It seems that in the presence of DNCs in alkaline solutions, two competitive processes are involved: polyhydroxy compound complexation in the following order of metal ion binding affinity, Ca(II) > Sr(II) > Ba(II), and inhibition on the onset of gold oxide formation in the same order. This last effect leads to an increased activity of the electrode surface; at the optimized value of detection potential of d-sorbitol, E(DET) = +50 mV vs Ag|AgCl, which is ~100 mV lower in comparison to the maximum value observed with blank carrier electrolytes (i.e., 0.58 M NaOH), there is an increase in sensitivity of ~50%, and 30% in the presence of 1.0 mM Sr(II), and 1.0 mM Ba(II), respectively. However, the current response of sample molecules results increased only when Ba(II) or Sr(II) ions were already contained in the alkaline media, that is the experimental condition normally occurring in flow injection and liquid chromatography systems. The voltammetric response, observed upon additions of Ba(II) or Sr(II) to an alkaline electrolyte containing d-sorbitol, showed progressive decrease of the anodic current. Irrespective of the experimental condition adopted, i.e., cation addition to the solution before or after the carbohydrate, the presence of Ca(II) has an adverse effect on the anodic currents. These findings have been explained by a rapid formation of adducts between polyhydroxy compounds and DNCs in sodium hydroxide solutions. PMID- 21639158 TI - Sample preconcentration using ion-exchange polymer film for laser ablation sampling inductively coupled plasma atomic emission spectrometry. AB - An efficient sample pretreatment/introduction technique for the inductively coupled plasma atomic emission spectrometry (ICP-AES) using ion exchange for analyte preconcentration and matrix separation and laser ablation sampling for sample introduction has been developed. Ammonium pyrrolidine dithiocarbamate (APDC)-polystyrene films are coated on glass plates for analyte preconcentration. Repetitive laser ablation sampling of the polymer film removes the ion-exchanged metal ions from the polymer film as fine particles for sample introduction into the ICP. After immersing the sample probe in a sample solution for 5 min, the ICP emission intensity for laser ablation of the polymer film is a few times larger than that after solution nebulization. The sample probe removes only a small fraction of the sample solution and, therefore, in principle, does not disturb the original solution significantly. Single-pulse laser ablation of the polymer film shows that the ion-exchanged metal ion concentration in the film reduces exponentially with the depth of the polymer film. Ion exchange to the polymer film is probably limited by the rate of metal ion diffusion into the film. Calibration curves for Cu, Hg, Pb, and Zn show linear dynamic range of ~1-2 orders of magnitude. The linear dynamic range for Cu increases to >3 orders of magnitude when using Pb as an internal standard. RSD of the ICP emission intensity is ~8%. PMID- 21639159 TI - Biochemical detection for direct bead surface analysis. AB - A continuous-flow biochemical detection system is presented which recognizes biologically active compounds immobilized to solid phases. This approach can be used to screen, for example, solid-phase combinatorial libraries for lead compounds. Biochemical detection is performed by mixing a plug of a solid-phase suspension with labeled affinity protein. During a short reaction time, the labeled affinity protein will only bind to ligands, i.e., compounds with biological activity. Hereafter, the free and bound labels are separated by means of a hollow fiber module. Quantitation of the free label is performed with a conventional flow-through fluorescence detector. Total assay time amounts to less than 3 min. Biochemical detection for direct bead surface analysis was developed for two model systems. The first model system used fluorescence-labeled avidin as affinity protein and its ligands biotin and iminobiotin immobilized to agarose as analytes. The second model system used fluorescence-labeled antisheep (Fab)(2) fragments as affinity protein and different IgGs immobilized to agarose as analytes. The feasibility of this approach for recognition of solid-phase immobilized ligands was documented by screening 50 samples with a 100% hit rate. PMID- 21639160 TI - Derivatization of steroids with dansylhydrazine using trifluoromethanesulfonic Acid as catalyst. AB - A new dansylation reaction, where trifluoromethanesulfonic acid is used as catalyst, has been characterized for six ketosteroids by employing experimental design followed by multivariate data analysis. The molar ratio between the steroid and the derivatization reagent was found to be the factor most strongly affecting the reaction. Faster reaction kinetics was achieved when the molar ratio between dansylhydrazine and the steroid was increased. Mass spectroscopic analysis showed that the dual peaks observed when derivatized progesterone was separated on an octadecyl silica stationary phase were due to the syn and anti hydrazones formed. We furthermore conclude that the dansylation reaction is subject to alkyl catalysis rather than acid catalysis, since methyl trifluoromethanesulfonate showed a strong catalytic action, while the catalytic action of trifluoromethanesulfonic acid was lower when diluted in other alcohols and disappeared in aprotic solvents. A sensitivity to water in the reaction mixture strengthens the evidence for alkyl catalysis. When optimal experimental conditions were used, derivatization of picomole amounts of ketosteroids could be accomplished in 25 min. Analysis of spiked plasma containing 0.2-2.0 nmol each of progesterone and 3alpha-hydroxy-5beta-pregnan-20-one showed overall recoveries of 69-76% and 40-55%, respectively. The corresponding 3sigma detection limits estimated from calibration curve data were 12 and 15 pmol (n = 4, 500 MUL injected). PMID- 21639161 TI - Quantitative Determination of Semivolatile Organic Compounds in Solution Using Trap-and-Release Membrane Inlet Mass Spectrometry. AB - This paper discusses the use of trap-and-release membrane inlet mass spectrometry (T&R-MIMS) for the quantitative determination of semivolatile organic compounds in real samples. We found that the T&R-MIMS technique is particular sensitive to relatively polar, semivolatile organic compounds. For example, the detection limits for the acids acetylsalicylic acid and phenoxyacetic acid were lowered by a factor of 100 as compared with those possible with standard MIMS, and caffeine was detectable only with the T&R-MIMS method. The detection limits were in the parts-per-billion range, and the dynamic range was 3 orders of magnitude. As a practical example of the application of the T&R-MIMS technique, we used it for the quantitative analysis of caffeine in ground coffee and tea leaves. Good agreement between T&R-MIMS and HPLC determinations was found, and the reproducibility of the whole analytical system for caffeine determination (extraction procedure and T&R-MIMS determination) was within 10% as relative standard deviation. However, for coffee, a large background from the essential oils prevented low-level work, such as the determination of residual caffeine in decaffeinated coffee. Obviously, the analysis of many complex matrixes will require the use of tandem mass spectrometry. PMID- 21639162 TI - Determination of alkylbenzenesulfonates in recent sediments by gas chromatography/mass spectrometry. AB - Alkylbenzenesulfonates together with soap are the most widely used anionic surfactants. Linear alkylbenzenesulfonates (LAS) were introduced in the mid-1960s as substitutes for the poorly biodegradable tetrapropylenebenzenesulfonates (TPS). A method is presented for the selective and quantitative determination of LAS and TPS in recent sediments. Alkylbenzenesulfonates were extracted from sediments using methanol. The methanolic extract was passed through a strong anionic exchange column. The alkylbenzenesulfonates contained in the acidic eluate were then derivatized to their corresponding trifluoroethyl esters and quantitatively determined by gas chromatography/mass spectrometry using positive chemical ionization. Limits of quantitation for 10 g sediment samples varied between 1.5 and 21 MUg/kg of dry sediment for single LAS isomers and between 71 and 220 MUg/kg for total LAS. Limits of quantitation for the total of TPS were at ~200 MUg/kg. Relative standard deviations of replicate analyses typically ranged from 5 to 10%. Recovery rates of LAS in spiked sediment samples ranged from 79 to 113%. The presented method was applied to surface and subsurface sediments also containing long-chain (C(14)-C(16))-LAS and mixtures of LAS and TPS. PMID- 21639163 TI - Characterization of arsenosugars of biological origin using fast atom bombardment tandem mass spectrometry. AB - A fast atom bombardment (FAB) mass spectrometric method has been developed for characterizing arsenosugar compounds. These compounds are of particular interest not only because their biochemistry requires further investigation but also because they are present at relatively high concentrations in commercial seaweed food products. FAB was used for the efficient generation of gas-phase ions of the various arsenosugar compounds. Negative-ion detection of such ions was found to be more sensitive than positive-ion detection due to the presence of negatively charged substituents. Negative-ion collision-induced dissociation (CID) tandem mass spectrometry (MS) of the [M - H](-) precursor ions results in the formation of numerous characteristic product ions via charge-remote fragmentation. These product ions provide abundant structural information for arsenosugar characterization. Separation of the arsenosugar-originating precursor ions from the matrix ions, always present in FAB mass spectra, is achievable using an analyzer resolution of 3000. This resolution allows for accurate selection of a precursor ion for subsequent CID experiments. However, by switching to a resolution of 1000, the quality of the tandem mass spectra can be slightly improved. Such an improvement is especially useful when analyzing nanogram amounts of arsenosugars. Furthermore, it was demonstrated that positive-ion tandem MS provides complementary information for the structural characterization of the arsenosugars examined. The mass spectrometric procedures developed in this study were further applied for the characterization an arsenosugar present in a partially purified algal (Sargassum lacerifolium) extract. PMID- 21639164 TI - A Quantitative Method for Determination of Lactide Composition in Poly(lactide) Using (1)H NMR. AB - A method has been developed to quantitatively determine the composition of d lactide and meso-lactide stereoisomer impurities in poly(lactide) containing predominantly l-lactide. In this method, the stereosequence information obtained from a few well-resolved resonances in the (1)H NMR spectrum representing RR and R stereogenic defects is used. The d-lactide and meso-lactide as minor components lead to RR and R stereogenic defects, respectively, which influence the isotactic chain length distribution and hence affect the polymer properties. Analytical equations relating the stereosequence probability to the lactide feed composition are not available due the complicated kinetics involved for the melt polymerization; viz. the preference for syndiotactic lactide addition decreases with reducing residual lactide concentration in the batch process. Hence, empirical correlations were determined by least-squares fit to the predictions for the specific stereosequence probabilities provided by Monte Carlo calculations of a number of lactide stereocopolymerizations. The Monte Carlo calculations simulate the kinetics observed for melt polymerization at 180 degrees C catalyzed by Sn(II) bis(2-ethylhexanoate) (Sn(II) octoate) in a 1:10 000 catalyst/lactide ratio. PMID- 21639165 TI - Application of wavelet transform to extract the relevant component from spectral data for multivariate calibration. AB - An approach aiming at extracting the relevant component for multivariate calibration is introduced, and its performance is compared with the "uninformative variable elimination" approach and with the standard PLS method for the modeling of near-infrared data. The extraction of the relevant component is carried out in the wavelet domain. The PLS results on these relevant features are better, and therefore, it seems that this approach can successfully be used to remove noise and irrelevant information from spectra for multivariate calibration. PMID- 21639166 TI - A Combined Chemical and Electrochemical Approach Using Bis(trifluoroacetoxy)iodobenzene and Glucose Oxidase for the Detection of Chlorinated Phenols. AB - A novel electrocatalytic approach using a chemical reaction and an enzymatic reaction has been developed for the measurement of 18 chlorophenol congeners, including highly chlorinated pollutants such as pentachlorophenol, 2,3,5,6 tetrachlorophenol, 2,3,4,6-tetrachlorophenol, and several trichlorophenols. Chlorophenols were oxidized to chlorobenzoquinones with very high yields using bis(trifluoroacetoxy)iodobenzene in 0.1 M trichloroacetic acid, pH 1.5, at ambient temperature. UV-visible spectrophotometry, cyclic voltammetry, and HPLC have been used to characterize the reaction products and yields. Together with glucose oxidase immobilized on a working glassy carbon electrode (+0.45 V vs Ag/AgCl), chlorinated benzoquinones have been demonstrated to be efficient mediators in a glucose oxidase/glucose system. In this approach, glucose oxidase was readily reduced by excess glucose to provide a non-rate-limiting source of electron flow toward the electrode. The oxidation products of chlorophenols then recycled the reduced glucose oxidase to its active oxidative state, i.e., mediating the rate-limiting electron transfer from the enzyme to the electrode. At pH 3.5, linear behavior of the current response was observed up to 200 nM for all chlorophenol oxidation products. The detection limit of this method for both pentachlorophenol and 2,3,5,6-tetrachlorophenol was about 4 nM, which is close to the maximum allowable contamination level of pentachlorophenol in water samples (2.7 nM). The detection limit obtained for pentachlorophenol could also be considered superior to the result obtained with the PCP immunoassay technology (13.3 nM). PMID- 21639167 TI - Surface mass spectrometry of biotinylated self-assembled monolayers. AB - Biotin and biotinylated self-assembled monolayers (SAMs) on gold have been investigated using time-of-flight secondary ion mass spectrometry, direct laser desorption, laser desorption with 193 nm photoionization of ion- and laser desorbed species, and laser desorption with vacuum ultraviolet (VUV, 118 nm) photoionization. Our results indicate that direct laser desorption and laser desorption combined with 193 nm multiphoton ionization can detect a chromophoric molecule like biotin that is covalently bound to a SAM. However, secondary ion mass spectra were dominated by fragmentation, and ion desorption/193 nm photoionization detected no species related to biotin. The dominant features of the laser desorption/VUV mass spectra were neat and Au-complexed dimers of intact and fragmented biotinylated SAM molecules. Multiphoton and single-photon ionization of laser-desorbed neutrals from biotinylated SAMs both led to the production of ions useful for chemical analysis of the monolayer. Multiphoton ionization with ultraviolet radiation was experimentally less challenging but required a chromophore for ionization and resulted in significant fragmentation of the adsorbate. Single-photon ionization with VUV radiation was experimentally more challenging but did not require a chromophore and led to less fragmentation. X-ray photoelectron spectra indicated that the biotinylated SAM formed a disordered, 40-60 A thick monolayer on Au. Additionally, projection photolithography with a Schwarzschild microscope was used to pattern the biotinylated SAM surface and laser desorption/photoionization was used to detect biotinylated adsorbates from the ~10 MUm sized pattern. PMID- 21639168 TI - Chemometric labeling of cereal tissues in multichannel fluorescence microscopy images using discriminant analysis. AB - This paper presents a novel, semiautomatic method for microscopic identification of multicomponent samples, which allows the identification, location, and percentage quantity of each component to be determined. The method involves applying discriminant analysis to a sequence of multichannel fluorescence microscopy images via a supervised learning approach; by selecting groups of pixels that are representative for each component type in a "known" sample, a computer is "taught" how to recognize the behavior (i.e., fluorescence emission) of the various components when illuminated under different spectral conditions. The identity, quantity, and location of these components in "unknown" samples (i.e., samples with the same component types but in different ratios or distributions) can then be investigated. The technique therefore enables semiautomatic quantitative fluorescence microscopy and has potential as a quality control tool. This work demonstrates the application of the technique to artificial and natural samples and critically discusses its quality, potential, and limitations. PMID- 21639169 TI - Strategies and data precision requirements for the mass spectrometric determination of structures from combinatorial mixtures. AB - Mass spectrometric data can be obtained for compounds in bead-bound combinatorial mixtures by several techniques. However, little specific information is available regarding (1) how well these data differentiate between candidate structures in large combinatorial pools, (2) what precision of data is required to achieve adequate specificity in these analyses, and (3) what are the best strategies for applying these data. In this work, computer modeling is used to address these questions. Strategies employing multiple filters (i.e., those that differentiate possible structures using more that one measured mass spectral parameter) are found to provide better specificity and to be more robust (that is, the specificity is less dependent on the precision of the data) than discrete filters. With moderate precision data (e.g., 50 ppm mass precision, 10% isotope ratio precision), multiple filter strategies are found to give unequivocal results for ~80% of the populations of combinatorial mixtures with most of the remaining degeneracy at the 2-fold level. A simple protocol for the application of multiple filter methods is presented. PMID- 21639170 TI - Determination of delta(18)O and delta(15)N in Nitrate. AB - The analyses of both O and N isotopic compositions of nitrate have many potential applications in studies of nitrate sources and reactions in hydrology, oceanography, and atmospheric chemistry, but simple and precise methods for these analyses have yet to be developed. Testing of a new method involving reaction of potassium nitrate with catalyzed graphite (C + Pd + Au) at 520 degrees C resulted in quantitative recovery of N and O from nitrate as free CO(2), K(2)CO(3), and N(2). The delta(18)O values of nitrate reference materials were obtained by analyzing both the CO(2) and K(2)CO(3) from catalyzed graphite combustion. Provisional values of delta(18)O(VSMOW) for the internationally distributed KNO(3) reference materials IAEA-N3 and USGS-32 were both equal to +22.7 +/- 0.50/00. Because the fraction of free CO(2) and the isotopic fractionation factor between CO(2) and K(2)CO(3) were constant in the combustion products, the delta(18)O value of KNO(3) could be calculated from measurements of the delta(18)O of free CO(2). Thus, delta(18)O(KNO)((3)) = adelta(18)O(free)( )(CO)((2)) - b, where a and b were equal to 0.9967 and 3.3, respectively, for the specific conditions of the experiments. The catalyzed graphite combustion method can be used to determine delta(18)O of KNO(3) from measurements of delta(18)O of free CO(2) with reproducibility on the order of +/-0.20/00 or better if local reference materials are prepared and analyzed with the samples. Reproducibility of delta(15)N was +/-0.10/00 after trace amounts of CO were removed. PMID- 21639171 TI - Genetic programming: a novel method for the quantitative analysis of pyrolysis mass spectral data. AB - A technique for the analysis of multivariate data by genetic programming (GP) is described, with particular reference to the quantitative analysis of orange juice adulteration data collected by pyrolysis mass spectrometry (PyMS). The dimensionality of the input space was reduced by ranking variables according to product moment correlation or mutual information with the outputs. The GP technique as described gives predictive errors equivalent to, if not better than, more widespread methods such as partial least squares and artificial neural networks but additionally can provide a means for easing the interpretation of the correlation between input and output variables. The described application demonstrates that by using the GP method for analyzing PyMS data the adulteration of orange juice with 10% sucrose solution can be quantified reliably over a 0-20% range with an RMS error in the estimate of ~1%. PMID- 21639172 TI - A multiresidue derivatization gas chromatographic assay for fifteen phenolic constituents with mass selective detection. AB - We have developed a GC/MS method to simultaneously measure the concentrations of 15 biologically active phenolic components of wine: vanillic acid, gentisic acid, m- and p-coumaric acid, gallic acid, ferulic acid, caffeic acid, cis- and trans-resveratrol, epicatechin, catechin, morin, quercetin, and cis- and trans polydatin. Wine (1 mL) was diluted 1:1 with water to reduce the alcohol content and extracted on a preconditioned C-8 solid-phase extraction cartridge. The phenolic compounds were eluted with ethyl acetate, evaporated to dryness, and derivatized with bis(trimethylsilyl)trifluoroacetamide/pyridine. The TMS derivative of each phenolic compound was analyzed on a GC/MSD coupled to a DB-5HT capillary column using one target and two qualifying ions for each compound in a total run time of 26 min. Resolution and quantitation of all compounds were excellent, with linear calibration curves over a wide range. The lowest detection limit was for gentisic acid (24 MUg/L) and highest for quercetin (843 MUg/L). The average percent recovery and coefficient of variation (mean precision) ranged from 90.7 to 104.6 (except morin, 72.2%) and 4.0 to 10.2 (except morin, 16.1%, and quercetin, 16.0%) respectively. This method has been applied to solid vitaceous plant materials as well as wine and should be suitable to measure polyphenols in fruit, vegetables, and other foods provided that efficient extraction techniques are employed. PMID- 21639173 TI - Determination of effective mobilities and chiral separation selectivities from partially separated enantiomer peaks in a racemic mixture using pressure-mediated capillary electrophoresis. AB - A new measurement principle has been developed for the rapid determination of the effective mobilities of enantiomers from their partially separated peaks. The method involves (i) partial separation of the enantiomers of a racemic sample by electrophoresis, (ii) pressure mobilization of the partially separated band by the detector, (iii) calculation of the effective separation distance between the enantiomer centroids from the observed band width and the extent of theoretical band broadening, and (iv) calculation of the effective mobilities of the enantiomers (and the separation selectivity) from the effective separation distance. Experimental conditions that lead to negligible nonideal ionic contributions to the band width are outlined. The proposed method eliminates the error caused by the changing electroosmotic flow, yields complexation constant and ionic mobility values that are more precise than the conventionally obtained ones, and reduces the measurement time by 80-90%. The equations required for the calculations are presented in a simple, ready-to-use spreadsheet format. PMID- 21639174 TI - A Family of Single-Isomer Chiral Resolving Agents for Capillary Electrophoresis. 2. Hepta-6-sulfato-beta-cyclodextrin. AB - A new, hydrophilic, single-isomer charged cyclodextrin, the sodium salt of hepta 6-sulfato-beta-cyclodextrin has been synthesized, characterized, and used for the capillary electrophoretic separation of the enantiomers of numerous noncharged, acidic, basic, and zwitterionic analytes. Hepta-6-sulfato-beta-cyclodextrin proved to be a much stronger complexing agent for all the analytes tested, in both low-pH and high-pH background electrolytes, than the previously synthesized, moderately hydrophobic heptakis(2,3-diacetyl-6-sulfato)-beta-cyclodextrin. The separation selectivities of the two single-isomer, differently functionalized charged cyclodextrins often proved to be complementary. In agreement with the predictions of the charged resolving agent migration model, separation selectivity for the noncharged analytes decreased as the concentration of hepta-6 sulfato-beta-cyclodextrin was increased. For acidic, basic, and zwitterionic analytes, selectivity could increase, decrease, or pass a maximum, depending on the binding strength of the enantiomers and ionic mobilities of both the complexed and noncomplexed forms of the enantiomers. PMID- 21639175 TI - Capillary zone electrophoresis in nonaqueous solvents in the presence of ionic additives. AB - Capillary zone electrophoresis (CZE) in nonaqueous media and in the presence of ionic additives has been successfully applied to the determination of compounds that differ only slightly in their electrophoretic mobilities. Triazine herbicides of environmental interest were chosen as test compounds because they behave as very weak bases. CZE separation of these analytes (especially chlorotriazines) in aqueous solution is difficult due to the low pH required for their conversion into protonated cationic form (HA(+)). However, in mixed nonaqueous solvents, 50% (v/v) acetonitrile-methanol, the acid-base characteristics of these compounds are modified, yielding the protonated ionic species that is susceptible to migration when subjected to an electric field. A noteworthy increase in separation selectivity and resolution can be achieved by using ionic additives. Thus, in this mode of capillary zone electrophoresis, separation is based on ionic interactions between the charged analytes and the ionic additive present in the separation medium. These interactions contribute to enhancing mobility differences and to improving analyte separation. For the separation of chloro- and methylthiotriazines, 10 mM perchloric acid in 50% (v/v) acetonitrile-methanol and 20 mM SDS proved to be satisfactory, providing high resolution in short analysis times. The selectivity achieved was found to depend on the degree of association of the analyte with the ionic additive in the nonaqueous medium. This permits manipulation of the selectivity of the electrophoretic separations as a function of the type and concentration of the ionic additive and of the nature of the nonaqueous medium employed. PMID- 21639176 TI - Determination of accurate electroosmotic mobility and analyte effective mobility values in the presence of charged interacting agents in capillary electrophoresis. AB - A method for determining the accurate effective mobility value of an analyte in the presence of a charged interacting agent, such as a charged cyclodextrin, a micellar agent, a protein, or a DNA fragment that binds the traditional electroosmotic flow markers, is presented. Part of the capillary is filled with the charged interacting agent-containing background electrolyte; the other part is filled with the charged interacting agent-free background electrolyte. The analyte band is placed in the charged interacting agent-containing background electrolyte zone, while a neutral marker (electroosmotic flow marker) is placed in the adjacent charged interacting agent-free background electrolyte zone. The initial, preelectrophoresis distance between the analyte band and the neutral marker band is determined by pressure mobilizing the bands past the detector and recording the detector trace. Subsequently, by applying reverse pressure, the bands are moved back into the first portion of the capillary and a brief electrophoretic separation is carried out. Then, the bands are pressure mobilized again past the detector to obtain their final, postelectrophoresis distance. If (i) the neutral marker does not come into contact with the charged interacting agent and (ii) the analyte does not migrate out of the homogeneous portion of the charged interacting agent zone, the accurate effective electrophoretic migration distance of the analyte, corrected for bulk flow transport, can be determined. The actual electric field strengths in the different zones of the heterogeneously filled capillary can be calculated from the integral of the electrophoretic current and the conductivity of the charged interacting agent-containing background electrolyte measured in a separate experiment. Once the effective mobility of an analyte in the charged resolving agent-containing background electrolyte is determined by this method, the analyte becomes a mobility reference probe for that background electrolyte and can be used to calculate the bulk flow mobility in subsequent conventional CE separations utilizing the same charged interacting agent. The new method can also be used to probe the interactions of the charged interacting agents and the wall of the capillary. PMID- 21639177 TI - Development and evaluation of flexible empirical peak functions for processing chromatographic peaks. AB - This paper describes the results of developing and evaluating extremely flexible empirical peak-shaped functions for processing chromatographic peaks. The proposed peak functions were developed based on transformation of Gaussian function into two-step functions that separately describe the leading and trailing edges. The flexibility and capability of these models were achieved by the combination and empirical modifications of the leading and trailing edge functions. The flexibility of the models was evaluated by fitting them to 10 types of peak shapes generated by literature peak functions possessing asymmetry values from 1 to 2.8. Excellent fits were found between the proposed models and generated peak shapes, showing that the new peak functions are extremely flexible. Furthermore, the capability of the models to smooth noisy peaks was demonstrated by fitting them to noisy, exponentially modified Gaussian peaks with different noise levels (S/N ratio was ranged from 200 to 10). Finally, we conclude that the flexibility of these models can be used to establish "templates" to significantly aid in smoothing noisy peaks and peak deconvolution. PMID- 21639178 TI - Complete enantiomeric separation of phenylthiocarbamoylated amino acids on a tandem column of reversed and chiral stationary phases. AB - The enantiomeric separation of phenylthiocarbamoyl derivatives of amino acids (PTC-AAs) was studied on a series of reversed phase HPLC columns coupled to the chiral phase HPLC columns. First, the five chiral phases (native, 0.2, 3.3, 7.5 and 16.9 phenylcarbamoylated/beta-cyclodextrins, Ph/CD) were newly prepared by modification of beta-cyclodextrin with phenyl isocyanate and were examined for the enantiomeric separation of PTC-AAs. Among them, the 3.3Ph/CD phase gave the best enantiomeric separation (alpha >= 1.04). However, the separation of the individual PTC-AAs was not sufficient. Next, these separations were investigated on various reversed phase HPLC columns, and octyl silica was selected in terms of the suitability of the mobile phase adopted for the enantiomeric separation mentioned above. The effects of the column temperature, the ion-pairing reagent, and the final content of methanol were also studied on the tandem column of octyl silica and the 3.3Ph/CD phase. Under the best conditions (100 mM ammonium acetate (pH 6.5) containing 1 mM butanesulfonate with 0-40% methanol as the mobile phase), all the individual PTC-AAs were well separated within 150 min. The applicability of the method was demonstrated by the sequence/configuration analysis of a peptide containing a d-amino acid ([d-Thr(2)]leucine enkephalin Thr). PMID- 21639179 TI - Direct determination of oxygen by HPLC. 1. Basic principles of a sensitive and selective oxygen sensor. AB - The basic principles of a novel, versatile, sensitive, and selective oxygen sensing assay are presented in this paper. For the first time, liquid chromatography with electrochemical detection (at the hmde) has been used for the determination of oxygen. All factors concerning optimization of the chromatographic separation conditions and electrochemical detection with respect to direct determination of oxygen even in complex biological samples are discussed. Due to the combination of a chromatographic technique with amperometric detection, a high selectivity can be achieved. A direct and linear relationship between the oxygen concentration in the sample and the reduction current was verified in a large concentration range from saturation down to trace level oxygen concentrations. The novel oxygen-sensing assay provides a much higher sensitivity compared to conventional oxygen sensors. In principle, O(2) concentrations down to 4.5 * 10(-)(9) mol L(-)(1) O(2) (corresponding to a signal to-noise ratio of 3) can be detected. Precision was determined by repeated measurements (n = 6) of air-saturated solutions (2.5 * 10(-)(4) mol L(-)(1) O(2), 20 degrees C, 920 mbar) which yielded relative standard deviations of lower than 0.2%. PMID- 21639180 TI - Direct determination of oxygen by HPLC. 2. Chamber and sample application system for determination of o(2) at trace levels. AB - All oxygen measurement systems so far available are characterized by a lack of suitable precision and/or required limit of detection, which would be essential for a great variety of applications. In this paper, a novel oxygen chamber together with a completely new concept of sample application ("two-chamber-siphon technique") is presented which can be used in combination with the previously reported chromatographic oxygen sensor (part 1). This new oxygen-sensing assay exhibits several advantages in comparison to conventional oxygen measurement systems: e.g., the uncontrollable influence of the surrounding atmosphere as well as oxygen consumption and storage processes are excluded. For the first time, measurements of molecular oxygen below 1 * 10(-)(7) mol L(-)(1) can be performed. Reliable quantification of oxygen in liquids and also in gaseous and solid samples can be achieved with utmost sensitivity (LOD 4.9 * 10(-)(9) mol L( )(1) O(2) = 98 fmol of oxygen on column) and precision (RSD = 0.7%, n = 8). PMID- 21639181 TI - Sol-gel-derived gold composite electrodes. AB - The preparation, characterization, and analytical utility of sol-gel-derived gold composite electrodes is described. The new metal-ceramic electrodes are comprised of gold powder homogeneously dispersed in a modified silica matrix. They couple the favorable electron-transfer kinetics common to gold surfaces with the regeneration, bulk modification, and versatility features of sol-gel-derived composite materials. The voltammetric characteristics of the composite gold silica electrodes are explored and compared with conventional gold electrodes. Sol-gel-derived gold biosensors have been prepared by incorporating an oxidase enzyme within the sol-gel gold solution. Analogous thick-film enzyme strips, based on a new screen-printable gold biogel ink, have also been fabricated. To our knowledge, the above represent the first examples of metal-ceramic sensing electrodes and of bulk modification of metallic working electrodes. PMID- 21639182 TI - Matrix representation of solution mixing by aliquot exchange. AB - We present a mathematical description of mixing two solutions by exchanging aliquots back and forth between them. We propose that this method of aliquot exchange can be used to automate calibration curve preparation in a way that produces less solvent waste than conventional serial dilution methods. We also show its use in quickly mixing solutions. The process of aliquot exchange is represented mathematically by a 2 * 2 symmetric matrix, A, that is a function of the volume or percentage of liquid, p, that is exchanged. Each cycle of aliquot exchange is represented by operating the matrix, A, on the previous concentrations. That is, after n mixing cycles, the final concentrations (C(i)(n)()) are given by A(p(n))A(p(n)(-1))...A(p(1)) operating on the initial concentrations (C(i)(0)), or (A(p))(n)()C(i)(0) if the same amount of liquid is exchanged in each step. We observe close agreement between theory and experiment. For solutions that have equal initial volumes, both the matrix A(p) and the product of any number of such matrices that may have the same or different values of p have equal diagonal and equal off-diagonal elements (they are symmetric), the sum of the elements in any row or column sums to unity, and the operation of any of these matrices on a set of concentrations produces two new concentrations that sum to the same value as the sum of the initial concentrations. We follow the mixing process for two solutions of equal volume by plotting the matrix element A(12)(n)() of A(n)(), which approaches 0.5 as n increases. As expected, the larger the exchanged aliquot, the more quickly the solutions mix. By varying the fraction, p, that is exchanged, we show that it should be possible to produce a calibration curve with values that vary in concentration over at least 3 orders of magnitude from just two solutions. PMID- 21639183 TI - Real-Time Quantitative Analysis of Combustion-Generated Polycyclic Aromatic Hydrocarbons by Resonance-Enhanced Multiphoton Ionization Time-of-Flight Mass Spectrometry. AB - We have combined resonance-enhanced multiphoton ionization (REMPI) time-of-flight mass spectrometry with on-line flame sampling to determine the centerline concentrations of naphthalene, fluorene, and anthracene in a pure methane + oxygen/argon (1:5) diffusion flame. Naphthalene concentrations between 100 parts per billion by volume (ppbV) and 6 parts per million by volume (ppmV) and fluorene concentrations below 50 ppbV are determined using one-color REMPI on jet cooled samples extracted from the flame; anthracene concentrations in the 5-40 ppbV range are determined using two-color REMPI. The REMPI ion signals are converted to absolute concentrations in real time by performing gas-phase standard additions to the flame sample. Isomer-selective detection of larger polycyclic aromatic hydrocarbons, such as perylene and benzo[a]pyrene, is possible using the two-color REMPI approach. PMID- 21639184 TI - Controlling DNA Fragmentation in MALDI-MS by Chemical Modification. AB - Fragmentation has proven to be a major factor limiting accessible mass range, sensitivity, and mass resolution in the analysis of DNA by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Previous work has shown that this DNA fragmentation is strongly dependent on both the MALDI matrix and the nucleic acid sequence employed. Fragmentation is initiated by nucleobase protonation, leading to cleavage of the N-glycosidic bond with base loss, followed by cleavage of the phosphodiester backbone. In this study, asymmetric oligonucleotides incorporating cytidine and cytidine analogs such as 5-methyl-2' deoxycytidine, 5-bromo-2'-deoxycytidine, aracytidine, and 2'-fluorodeoxycytidine nucleosides were used to systematically investigate the influence of the structural changes on the stability of the N-glycosidic bond. Modifications of the deoxyribose sugar ring by replacing the 2'-hydrogen with more electron withdrawing groups such as the hydroxyl or fluoro group stabilize the N glycosidic bond to a greater extent than the C5 nucleobase modifications. 2' Hydroxyl and 2'-fluoro groups respectively are shown to partially or completely block fragmentation at the modified nucleosides. Mixtures of oligonucleotides incorporating such modifications demonstrate remarkably extended accessible mass range, as well as increased sensitivity and mass resolution. The stabilization provided by these chemical modifications also expands the range of matrices useful for nucleic acid analysis, yielding in some cases greatly improved performance. PMID- 21639185 TI - Vacancy capillary zone electrophoresis and differential capillary zone electrophoresis: variations on a well-known theme. AB - Two new variants on capillary zone electrophoresis (CZE) are described and experimentally investigated. In vacancy CZE the sample, diluted with a background electrolyte, fills the electrode vessels and the separation capillary. When pure background electrolyte is injected, the resulting electropherogram represents the composition of the sample. The electropherogram is almost identical with the result of a conventional CZE experiment. In differential CZE the sample again fills the electrode vessels and the separation compartment. After injection of a slightly different sample, a differential electropherogram is obtained that represents the differences between the two samples. The retention times of both new variants are comparable with conventional CZE, the separation efficiency, in terms of theoretical plates, is marginally lower, and both show good quantitative characteristics. The concept of vacancy electrophoresis explains the origin of unexpected system peaks in conventional CZE when multiple co-ions in the background electrolyte are used. PMID- 21639186 TI - Capillary Electrokinetic Chromatography Employing p-(Carboxyethyl)calix[n]arenes as Running Buffer Additives. AB - Calixarenes, a class of macrocyclic phenolic compounds with a basket-like shape, are used as capillary electrophoresis reagents for separations of native and substituted polycyclic aromatic hydrocarbons. The p-(carboxyethyl)calix[n]arenes reported herein are a series of charged, moderately water soluble macrocyclic molecules that can form complexes with neutral molecules. Electrokinetic chromatographic separations are based on the differential distribution of molecules between a running buffer phase, which is transported by electroosmotic flow, and an electrophoretically mediated calixarene. The size of the calixarene influences separation performance, illustrating the importance of cavity size and geometry in the complexation process. p-(Carboxyethyl)calix[7]arene provides the best efficiency (>10(5) plates/m) and selectivity in these studies. The influences of pH, organic solvent, and field strength on elution range, capacity factors, efficiency, and selectivity are also reported. In general, capacity factors are rather low, but the high charge-to-mass ratios of certain calixarenes produce relatively wide elution ranges. Molecular modeling data and solubility data are used to interpret the observed selectivity. PMID- 21639187 TI - Colloid characterization by sedimentation field-flow fractionation: correction for particle-wall interaction. AB - The effect of particle-wall interaction on retention ratios in sedimentation field-flow fractionation (FFF) is shown to be describable in terms of a semiempirical parameter delta(w) having units of length. A method of experimentally determining the value of this parameter for a given system is presented. It requires the determination of retention ratios for a set of particle standards over a range of field strengths. The value of delta(w) is simply obtained from the slope of a certain straight line plot of the retention data. Once determined, this parameter may be incorporated into the procedure for the determination of particle diameters from measured retention times to take account of the effects of particle-wall interaction. In principle, delta(w) is independent of field strength and is the same for different FFF instruments providing they employ the same carrier liquid and channel wall material. Therefore, delta(w) values have the potential to be universal system constants transferable from one instrument to another. PMID- 21639188 TI - Effect of electrical double-layer overlap on the electroosmotic flow in packed capillary columns. AB - A plug electroosmotic velocity profile is generally assumed to be characteristic of capillary electrochromatography. However, this ideal plug flow may be illusive in some experiments with packed-capillary columns due to overlap of electrical double layers in flow channels. We report here a theoretical analysis of the double-layer overlap effects in packed-capillary columns, which is based on Rice and Whitehead's theory of electroosmotic flow combined with a capillary tube model for porous packing. The results show that the electroosmotic velocity under the influence of double-layer overlap depends strongly on the operating parameters, which increases with the column porosity, the particle diameter, and the electrolyte concentration. PMID- 21639189 TI - Calibration of a commercial solid-phase microextraction device for measuring headspace concentrations of organic volatiles. AB - Solid-phase microextraction (SPME) is a versatile new technique for collecting headspace volatiles prior to GC analysis. The commercial availability of uniform SPME fibers makes routine, practical quantitation of headspace concentrations possible, but straightforward information for relating GC peak areas from SPME analyses to headspace concentrations has not been available. The calibration factors (amount absorbed by the fiber divided by headspace concentration) were determined for 71 compounds using SPME fibers with a 100 MUm poly(dimethylsiloxane) coating. The compounds ranged from 1 to 16 carbons in size and included a variety of functional groups. Calibration factors varied widely, being 7000 times higher for tetradecane than for acetaldehyde. Most compounds with a Kovats retention index of <1300 on a nonpolar GC column (DB-1) equilibrated with the fiber in 30 min or less. A regression model is presented for predicting the calibration factor from GC retention index, temperature, and analyte functional class. The calibration factor increased with retention index but decreased with increasing sampling temperature. For a given retention index, polar compounds such as amines and alcohols were absorbed by the fibers in greater amounts than were hydrocarbons. Henry's law constants determined using SPME were in general agreement with literature values, which supported the accuracy of the measured calibration factors. An unexpected concentration dependence of calibration factors was noted, especially for nitrogen-containing and hydroxy compounds; calibration factors were relatively higher (the SPME fiber was more sensitive) at the lower analyte concentrations. PMID- 21639190 TI - Selectivity in capillary electrophoresis: application to chiral separations with cyclodextrins. AB - In order to accurately evaluate the performances of any electrolyte medium, a clear concept of selectivity in capillary electrophoresis and related electroseparation techniques is proposed. Selectivity is defined as the ratio of the affinity factors of both analytes for a separating agent (phase, pseudophase, or complexing agent present in the background electrolyte). When in the presence of a complexing agent and if only 1:1 complexation occurs, selectivity corresponds to the ratio of the apparent binding constants and is independent of the concentration of the complexing agent. This concept is illustrated through the separations of neutral and anionic enantiomers in the presence of a cationic cyclodextrin, the mono(6-amino-6-deoxy)-beta-cyclodextrin, as a chiral complexing agent. The values obtained for different pairs of enantiomers are discussed with regard to the functional groups that distinguish them. When the analytes have the same mobilities in free solution and in their complexed form, then the resolution equation developed in micellar electrokinetic chromatography may be applied and optimum conditions (affinity factors, chiral agent concentration) can be predicted. PMID- 21639191 TI - Intrinsic selectivity in capillary electrophoresis for chiral separations with dual cyclodextrin systems. AB - Defined as the ratio of the affinity factors of the analytes for a complexing agent, the intrinsic selectivity is representative of the very nature of the complexing agent. When more than one complexing agent are present in the background electrolyte, it is possible to define several intrinsic selectivities according to whether complexing agents are considered separately or all together. A theoretical model with respect to selectivity is presented for separations that involve two complexing agents, using the concept of apparent constant for complex formation. When only independent complexation occurs (absence of mixed complexes), then the intrinsic selectivity of a complexing agent X in the presence of a complexing agent Y can be easily related to the intrinsic selectivity of each complexing agent and to complex formation constants. Dual systems of cyclodextrins (CDs), implementing the cationic mono(6-amino-6-deoxy) beta-cyclodextrin (beta-CD-NH(2)) and a neutral CD (trimethyl-beta-CD (TM-beta CD) or dimethyl-beta-CD (DM-beta-CD)), were studied to illustrate this model and to offer an alternative to the separation of neutral enantiomers when beta-CD NH(2) shows no or insufficient stereoselectivity. With a dual beta-CD-NH(2)/TM beta-CD system at pH 2.3, arylpropionic acid enantiomers were baseline resolved and benzoin derivatives were partially resolved. For the arylpropionic acids, beta-CD-NH(2), which is not stereoselective, confers on them a nonzero mobility, while TM-beta-CD allows the chiral recognition. A study of the respective influence of TauM-beta-CD and beta-CD-NH(2) concentrations was performed to determine the optimal conditions with respect to resolution. This theoretical approach allowed characterization of the intrinsic selectivity of neutral CDs for pairs of neutral enantiomers and therefore identification of the potential of neutral chiral agents for neutral enantiomers. PMID- 21639192 TI - Estimation of air/coating distribution coefficients for solid phase microextraction using retention indexes from linear temperature-programmed capillary gas chromatography. Application to the sampling and analysis of total petroleum hydrocarbons in air. AB - The paper describes a method to quantify hydrocarbons in air exclusively on the basis of chromatographic parameters without the need for calibration. A simple technique is presented to estimate distribution coefficients (K) between air and the poly(dimethylsiloxane) solid phase microextraction (SPME) fiber coating using the linear temperature-programmed retention index system (LTPRI). There is a linear relationship (r(2) = 0.99989) between the log K for a series of n-alkanes and LTPRI, thus providing a means by which establishing a K for any peak in a chromatogram is possible given its published or experimentally determined LTPRI. This alternative approach to establishing K values significantly enhances and simplifies the use of SPME for sampling and analyzing air for quantification of compounds without the need for fiber calibration. Analysis of a group of 29 isoparaffinic compounds and a group of 33 aromatic compounds showed excellent agreement between their theoretical air to fiber distribution coefficients based on LTPRI and the experimentally obtained distribution coefficients. In addition, for a very complex mixture of organics such as gasoline, SPME can establish a total petroleum hydrocarbon in air level using LTPRI. This method was carefully evaluated, and the results were essentially identical between standard procedures and the proposed simple procedure described in the paper. PMID- 21639193 TI - Effect of mobile phase additives in packed-column subcritical and supercritical fluid chromatography. AB - The effect of mobile phase additives is investigated for a variety of compounds under subcritical and supercritical conditions using packed columns. Retention of hydrogen bond donor/acceptor analytes was found to be more dependent on the presence of mobile phase additives than weak hydrogen bond acceptor analytes. The temperature and pressure of the mobile phase are major factors in the extent of this dependence. Consequently, selectivity between homologous compounds is dependent on both the additive used and the state of the mobile phase. Efficiency is nearly always improved by the presence of mobile phase additives, more so under supercritical conditions than under subcritical conditions. These observations suggest that surface molar excesses of mobile phase additives play a large part in the resulting character of the supercritical chromatographic system. PMID- 21639194 TI - Rate measurements by the pulsed-accelerated-flow method. AB - A pulsed-accelerated-flow spectrophotometer with UV-visible capability is described that permits measurement of pseudo-first-order rate constants as large as 500 000 s(-)(1) (t(1/2) = 1.4 MUs). Chemical rate processes are resolved from physical mixing rate processes by variation of flow velocities under conditions of turbulent flow. Two mixing processes are found in the mixing/observation tube. One mixing rate constant, valid for the full length of the tube, is directly proportional to the flow velocity. The other mixing behavior, proportional to the square of the flow velocity, is found only in the immediate vicinity of the 10 inlet reactant streams that collide with one another in the middle of the observation tube. Contributions from the latter mixing become more important for very fast reactions that take place close to the inlet jets. These mixing models and improved signal/noise permit the measurement of rate constants for very fast reactions. Applications of the PAF method to electron-transfer, proton-transfer, hydrolysis, and non-metal redox reactions are reported for pseudo-first-order and second-order reactions. PMID- 21639195 TI - Development of an amperometric detector for packed capillary column supercritical fluid chromatography. AB - Amperometric detection at a naked platinum microelectrode is shown to be compatible with pressure-programmed packed capillary column supercritical fluid chromatography (SFC) using carbon dioxide modified with as little as 1% acetonitrile. Amperometric detection in modified carbon dioxide is, hence, possible without addition of salts, which otherwise may limit the stability of the chromatographic system. The detection, which can been based on either oxidations or reductions, is also compatible with the use of methanol as a sole modifier. Picogram amounts of ferrocene could be detected after separations both at a constant pressure and under pressure-programmed conditions. The detection limit for ferrocene, at a constant pressure of 200 atm, was found to be approximately 20 pg. The response of the amperometric detector was observed to increase with decreased mobile phase density and increased amount of added modifier. The injection of increasing amounts of analytes resulted in increased peak tailing, most likely due to the limited solubility of the oxidation products in the mobile phase. The influence of this effect was, however, small for the amounts of analytes relevent in capillary column SFC. PMID- 21639196 TI - Limiting Currents for Steady-State Electrolysis of an Equilibrium Mixture, with and without Supporting Inert Electrolyte. AB - An exact treatment derives the steady-state limiting current of a one-electron reduction for the N ? O(+) + A(-) mixture at a hemispherical microelectrode. Either or both of the neutral N and cationic O(+) species may be electroactive. A supporting salt is present at any concentration, including zero or excess; its ions are electropassive and do not interact with the other solutes or each other. The various species are treated as having distinct diffusivities, linked to their mobilities through the Nernst-Einstein relationship. Universal electroneutrality is assumed. The predictions of the model are compared with published experimental data on the reduction of aqueous weak acids; agreement is excellent at intermediate, but poor at low, support ratios. Analysis of the unsupported case shows that the neutral N species dissociates in a narrow zone close to the electrode, and the injection of ions there serves to increase the electric field in the outer region of the transport zone. This enhances cationic migration enormously, leading to an unsupported limiting current that is much more than double the supported value. However, the limiting current is drastically diminished by traces of foreign electrolyte. Curiously, the limiting current with full support adopts the same value when equilibration is fast as when it is very slow, although the mechanisms are totally different. PMID- 21639197 TI - Amperometric sensors for simultaneous superoxide and hydrogen peroxide detection. AB - A two-channel sensor capable of almost instantaneous simultaneous detection of superoxide radical and hydrogen peroxide in the concentration range 10(-)(7)-10( )(4) M is very important for understanding of a number of rapid kinetics processes. A glassy carbon working microelectrode covered by an electrodeposited polypyrrole/horseradish peroxidase (PPy/HRP) membrane was employed as a H(2)O(2) sensor. Another glassy carbon microelectrode covered by a composite membrane of an inside layer of PPy/HRP and an outside layer of superoxide dismutase was employed as a working electrode for superoxide detection. These two working electrodes with Pt counter and tungsten oxide (WO(3)) reference electrodes were contained in one 6 mm diameter Teflon cylinder. Simultaneous measurements were performed at a potential of -60 mV (vs WO(3) reference, pH 5.1). Additional sensor characterization was performed for pH 5.1-9.0. Superoxide sensor behavior as a function of membrane deposition conditions and coating time is reported. Sensors' mutual influence, selectivity, response times, linearity, stability, and sensitivity for hydrogen peroxide and superoxide are presented and discussed. A mathematical model of sensors' responses is proposed, with model calculation corresponding to experiment within 10%. PMID- 21639198 TI - Application of a 32-microband electrode array detection system for liquid chromatography analysis. AB - An electrode array consisting of 32 microband electrodes and a generator electrode was constructed. With this electrode array, both the oxidation and reduction three-dimensional hydrodynamic chromatovoltammograms of a reversible reaction can be obtained in a single voltammetric run. The versatility of this electrode array was demonstrated by running solutions containing various phenolic compounds with an FIA and an HPLC system. From the oxidation and reduction chromatovoltammograms obtained, individual components in the sample solution can be determined quantitatively, even when overlapping of the chromatography peaks occurred. With this microband electrode array, detection limits down to 2.0 * 10( 8) M for the studied phenolic compounds were obtained. PMID- 21639199 TI - Nanoscale characterization of gold colloid monolayers: a comparison of four techniques. AB - Atomic force microscopy (AFM), field emission scanning electron microscopy (FE SEM), transmission electron microscopy (TEM), and near-field scanning optical microscopy (NSOM) have been used to characterize the nanostructure of Au colloid based surfaces. Because these substrates are composed of particles whose dimensions are known prior to assembly, they are well-suited for a critical comparison of the capabilities and limitations of each nanoscale imaging technique. The three criteria for this comparison, which are relevant to the field of nanoparticle assemblies in general, are (i) accuracy in establishing particle size, particle coverage, and interparticle spacing; (ii) accuracy in delineating surface topography; and (iii) ease of sample preparation, data acquisition, and image analysis. For colloidal Au arrays, TEM gives the most reliable size and spacing information but exhibits the greatest constraints with regard to sample preparation; in contrast, AFM is widely applicable but yields data that are the least straightforward to interpret. For accurate information regarding nanometer-scale architecture of particle-based surfaces, a combination of at least one scanning probe method (AFM, NSOM) and one accelerated-electron method (TEM, FE-SEM) is required. PMID- 21639200 TI - Particulate material analysis by a laser ionization fast conductivity method. Water content effects. AB - Combined laser multiphoton ionization and fast-conductivity methods have been applied to probe organic contamination on wet particulate samples. This is a first attempt of testing such a technique for this purpose. A special emphasis has been made on establishing correlation between sample water content and detection of pyrene, which has been used as a probe contaminant in this study. Environmental (soil) and artificial samples (silica gel) have been studied. The experimental setup includes a pulsed N(2) laser and a fast-conductivity detection system, operated in an opened chamber under ambient conditions. The correlation between the observed photoionization signals and water content has been investigated in a slow-drying mode, where water has been gradually evaporated. Surface contamination of wet samples has been more efficient than moist ones. This was evident, in the case of soil samples, from the earlier appearance (at lower pyrene concentrations) of saturation-type features of the detected photoionization signals. Assumptions of a contamination model has enabled certain compensation for sample and matrix effects. PMID- 21639201 TI - Supercritical fluid extraction of natural antioxidants from rosemary: comparison with liquid solvent sonication. AB - Supercritical fluid extraction (SFE) and liquid solvent sonication, in combination with two different sample treatments, were compared for the extraction of natural antioxidants from rosemary leaves. Dried, ground, and sieved rosemary leaves (20 mg) were subjected to SFE with CO(2) at 355 bar at 100 degrees C (CO(2) density 0.72 g/mL) for 20 min at a liquid flow rate of 4 mL/min. The analytes were concentrated on an ODS trap and subsequently eluted with acetone. Antioxidants in the SF and liquid solvent extract were analyzed by HPLC. Compounds of known antioxidant activity such as carnosol, carnosic acid, and methyl carnosate were identified by mass spectrometry of the HPLC fractions collected. Freezing and grinding the samples in liquid nitrogen resulted in decreased carnosic acid recoveries. Supercritical CO(2) extraction provided the highest recovery of carnosic acid from rosemary leaves (35.7 mg/g), the lowest relative standard deviation (4.4%), and the cleanest extract [Formula: see text] no cleanup prior to HPLC was required. Among the liquid solvents studies, only acetone provided comparable results (73% recovery relative to SC-CO(2) extraction); however, it required decoloration with active carbon prior to HPLC analysis. PMID- 21639202 TI - Minimization of transition metal interferences with hydride generation techniques. AB - A new hydride generator has been characterized for use with the acid-NaBH(4) hydride generation systems based on the insertion of a capillary tube into the sample introduction channel of a standard Meinhard nebulizer. The acidic sample and the tetrahydroborate solution are mixed at a merge point 1.5 cm from the end of the nebulizer orifice. Nebulization of the reaction solutions into a 0.7 mL tubular "spray chamber" follows a very short mixing time (less than 0.012 s) of the reagents. This approach permits 10 000 MUg/mL Ni(2+) or Cr(3+) to be present in the sample solution without producing any interferences. Additionally, in the presence of Fe(3+) added as a "releasing agent", 5000 MUg/mL Co(2+) or 160 MUg/mL Cu(2+) can also be tolerated without interference. An 80 +/- 2% generation efficiency is attained for the test element selenium. A detection limit of 6 MUg/L (3sigma(b)) is achieved with ICP-AES detection. Precision of replicate measurements at the 12 MUg/L level varies from 5 to 12% relative standard deviation. PMID- 21639203 TI - Temperature-resolved molecular emission spectroscopy: an analytical technique for solid materials. AB - Temperature-resolved molecular emission spectroscopy is described as a thermal analysis method for the analysis of solids and liquids. The technique uses an electrically heated graphite cup to decompose and/or vaporize the sample. The vapors are carried by a stream of argon into a cool hydrogen diffusion flame. Both the quantity and the nature of the decomposed species can be determined. The technique is particularly useful for the determination of sulfur, phosphorus, or nitrogen. Calibration curves for sulfur show the expected parabolic shape, and those for phosphorus are linear. The detection limit for elemental sulfur was determined to be approximately 50 ng. The evolution of sulfur is shown to be related to the decomposition temperature which is characteristic of the sulfur containing species. Reproducibility of the decomposition temperatures is typically +/-2%. PMID- 21639204 TI - Packed-Column Supercritical Fluid Chromatography: Quantitative Determination of Uranium without Liquid Waste Generation. AB - A new procedure for the determination of uranium by packed-column supercritical fluid chromatography is proposed. A nonfluorinated chelating agent selective for copper and uranium, the 2,6-diacetylpyridine bis(benzoylhydrazone), has been chosen. We have studied its chromatographic properties on different stationary phases and the influence of the methanol content in the carbon dioxide mobile phase. The separation of the metal compounds was conducted with and without solvent injection. A calibration curve was obtained for uranium in the range of 52-323 ng injected. The accuracy of the method is 0.5%, the repeatability 4%. The same studies were performed with a new compound, diacetyl-2,6 pyridine bis(4-tert butyl benzoylhydrazone). An increase in retention and efficiency was then observed. PMID- 21639205 TI - Molecular interactions at octadecylated chromatographic surfaces. AB - Interactions between fluorescent solutes and an octadecylated silica surface are investigated using fluorescence emission spectra and quenching techniques under conditions similar to those found in high-performance liquid chromatography. Pyrene, benzo[a]pyrene, fluorene, biphenyl, propyldansylamide, and decyldansylamide are used as the fluorescent probes, and potassium iodide and N,N dimethylaniline are used as quenchers. N,N-Dimethylaniline is a moderately retained quencher thought to probe deeper into the octadecylated surface than the ionic iodide salt. Solvent-dependent fluorescence emission maxima, solvent dependent fluorescence vibronic band intensities, and quencher access to the probes are investigated using aqueous methanol mobile phase compositions ranging from 60 to 100% methanol. The resulting information is used to interpret differences in interfacial probe environments and to determine the location of the probes within the bonded phase layer. The data indicate that biphenyl and pyrene may remain in very nonpolar interfacial probe environments deep in the bonded phase layer over the mobile phase composition range tested. The fluorophore portion of both propyldansylamide and decyldansylamide may reside in an interfacial environment, which becomes more polar as the water content in the mobile phase is increased. Benzo[a]pyrene apparently becomes exposed to the mobile phase as the water content in the mobile phase increases. This is thought to be due to the relatively large size of the solute molecule and the collapse of the octadecyl chains with increased solvent polarity. Fluorene appears to interact strongly with silanol groups. The results are interpreted in light of the surface convolution and chain cluster octadecylated chromatography surface models and are found to be more constant with the chain cluster model. Implications of the results to reversed phase high-performance liquid chromatography separations are also discussed. PMID- 21639206 TI - Temperature-dependent electron capture detector response to common alternative fluorocarbons. AB - The relative electron capture detector (ECD) response to alternative fluorocarbons (AFCs) using gas chromatography are found to be at least 1 order of magnitude lower than that for CFC-12. Detection limits for the chlorofluorocarbons CFC-12, HCFC-22, HCFC-123, and HCFC-124 are found to be 2.5, 90, 30, and 90 pg, respectively. Those for the hydrofluorocarbons are significantly poorer; 14 and 45 ng for HFC-125 and HFC-134a, respectively. HFC 152a was not detected using ECD. Since atmospheric concentrations of these compounds are in the low part-per-trillion level, GC-ECD is apparently not sensitive enough to be used for AFC analysis without substantial preconcentration. Two columns are evaluated for the AFC separation. The Poraplot Q WPLOT column showed good separation ability, though column bleed limits detection performance. A Carboxen 1004 packed column exhibits much lower interference. But separations are time consuming and peak broadening adversely affects limits of detection. Mechanisms for the ECD response are proposed based on thermodynamics and temperature-dependent ECD responses. CFC-12, HCFC-123, and HFC-125 apparently undergo ion-forming dissociative electron capture. The electron capture process for HCFC-22 and HFC-134a appear to form molecular ions. Both mechanisms appear to be operative for HCFC-124 electron capture. Dissociative electron capture rate constants for HCFC-123, HCFC-124, and HFC-125 are estimated to be 3.5 * 10(-)(10), 1.0 * 10(-)(10), and 5.6 * 10(-)(13) cm(3) s(-)(1), respectively at 300 degrees C. PMID- 21639207 TI - Prediction of chiral chromatographic separations using combined multivariate regression and neural networks. AB - A new method for the prediction and description of enantioselective separations on HPLC chiral stationary phases (CSPs) is described. Based on the combination of multivariate regression and neural networks, the method was successfully applied to the separation of a series of 29 aromatic acids and amides, chromatographed on three amylosic CSPs. Combinations of charge transfer, electrostatic, lipophilic, and dipole interactions, identified by multivariate regression, were found to describe retention and enantioselectivity, with highly predictive models being generated by the training of back-propagation neural networks. PMID- 21639208 TI - Sol-gel coating technology for the preparation of solid-phase microextraction fibers of enhanced thermal stability. AB - A novel sol-gel method is described for the preparation of solid-phase microextraction (SPME) fibers. The protective polyimide coating was removed from a 1-cm end segment of a 200 MUm o.d. fused-silica fiber, and the exposed outer surface was coated with a bonded sol-gel layer of poly(dimethylsiloxane) (PDMS). The chemistry behind this coating technique is presented. Efficient SPME-GC analyses of polycyclic aromatic hydrocarbons, alkanes, aniline derivatives, alcohols, and phenolic compounds in dilute aqueous solutions were achieved using sol-gel-coated PDMS fibers. The extracted analytes were transferred to a GC injector using an in-house-designed SPME syringe that also allowed for easy change of SPME fibers. Electron microscopy experiments suggested a porous structure for the sol-gel coating with a thickness of ~10 MUm. The coating porosity provided higher surface area and allowed for the use of thinner coatings (compared with 100-MUm-thick coatings for conventional SPME fibers) to achieve acceptable stationary-phase loadings and sample capacities. Enhanced surface area of sol-gel coatings, in turn, provided efficient analyte extraction rates from solution. Experimental results on thermal stability of sol-gel PDMS fibers were compared with those for commercial 100-MUm PDMS fibers. Our findings suggest that sol-gel PDMS fibers possess significantly higher thermal stability (>320 degrees C) than conventionally coated PDMS fibers that often start bleeding at 200 degrees C. This is due, in part, to the strong chemical bonding between the sol gel-generated organic-inorganic composite coating and the silica surface. Enhanced thermal stability allowed the use of higher injection port temperatures for efficient desorption of less-volatile analytes and should translate into extended range of analytes that can be handled by SPME-GC techniques. Experimental evidence is provided that supports the operational advantages of sol gel coatings in SPME-GC analysis. PMID- 21639209 TI - Characterization of an inductively coupled plasma with xylene solutions introduced as monodisperse aerosols. AB - The optimum operating parameters of the inductively coupled plasma (ICP) with organic solvents are different from those with aqueous solutions. With organic solvents, the ICP is operated at higher power to overcome plasma cooling due to the organic solvent, and aerosol desolvation is necessary in order to reduce solvent load into the plasma. The monodisperse dried microparticulate injector (MDMI) offers the possibility of controlling solvent load by controlling the frequency with which droplets are introduced into the plasma. A test solution that contained 0.5 mg/L Ba in xylene was used to study the influence of MDMI operating parameters on the behavior of the ICP with an organic solvent. The spatial and temporal distribution of the solvent in the ICP was determined for different droplet production frequencies and MDMI oven temperatures, and the optimum operating conditions were established. The best detection limit for Ba in xylene was 1.5 ng/mL, or 0.16 pg (in 200 droplets). PMID- 21639210 TI - Measurement of Fluorescence from Tryptophan To Probe the Environment and Reaction Kinetics within Protein-Doped Sol-Gel-Derived Glass Monoliths. AB - Optically clear, ultrathin monoliths that contained the single-tryptophan protein monellin were prepared by the sol-gel technique from tetraethyl orthosilicate (TEOS). Suitable precautions were established to eliminate background fluorescence from impurities in TEOS, scattering from the monoliths, and photobleaching of the entrapped protein. Fluorescence spectra and anisotropy results indicated that useful, essentially scatter-free fluorescence signals could be obtained from the intrinsic tryptophan residue of monellin which was entrapped into either wet-aged or dry-aged monoliths. The combination of spectral, quenching, and anisotropy results suggested that the mobility of solvent inside monoliths was substantially reduced compared to bulk solution, providing a possible explanation for the improvements in protein stability that occur upon entrapment. The monitoring of intrinsic protein fluorescence also provided information about the kinetics of the interaction between the entrapped protein and external reagents. The interaction of monellin with both neutral and charged species was examined under conditions of continuous stirring and indicated response times on the order of minutes. In the case of the neutral species, the kinetics were best described by a sum of first-order rate constants when the reactions occurred in the glass matrix. For charged species, interactions between the analytes and the negatively charged glass matrix caused the reaction kinetics to become complex, with the overall reaction rate depending on both the type of aging and the charge on the analyte. These findings suggest that caution must be exercised when entrapped proteins are used for sensing of charged species. PMID- 21639211 TI - Synthesis and solid state (13)c and (19)f NMR characterization of aliphatic and aromatic carboxylic Acid fluoride polymers. AB - A new chemical/spectroscopic couple that differentiates aromatic from aliphatic carboxylic acid polymers was developed. The method is complementary to more traditional methods of identification (IR) and is applicable to the analysis of complex mixtures where IR determinations are complicated by extensive vibrational band overlap. The method entails (1) conversion of carboxylic acids into acid fluorides and (2) specific detection of the resonances of the acid fluoride carbon and of the carbon directly attached to the carbonyl carbon by solid state (19)F-(13)C cross polarization (CP)/MAS (13)C NMR. The assignment of the chemical shift of the latter resonance to either the sp(2) or sp(3) carbon resonance manifold specifies the nature of the acid functionality. The preparation of the acid fluoride derivatives of several polymers containing aliphatic and aromatic carboxylic acid functionality was evaluated using sulfur tetrafluoride (SF(4)), diethylaminosulfur trifluoride (DAST), cyanuric fluoride, and thionyl fluoride. Room temperature reactions using DAST in methylene chloride or neat SF(4) gave the acid fluorides in yields >=90% for the acids studied. Aromatic acid fluoride yields were essentially quantitative. Aliphatic acid fluorides were contaminated with the anhydride of the acid. The acid fluoride polymers were characterized by solid state (13)C and (19)F MAS/NMR. PMID- 21639212 TI - Combined Ion Mobility/Time-of-Flight Mass Spectrometry Study of Electrospray Generated Ions. AB - A commercially available ion mobility spectrometer was interfaced to a custom built linear time-of-flight (TOF) mass spectrometer for the purpose of examining electrospray-generated plumes. Ionic species that were separated in the ion mobility spectrometer could be selectively determined with the TOF mass spectrometer. Tetraalkylammonium salts, a peptide, and proteins were examined. Their ion mobility spectra typically comprised a few peaks; some of these mobility-resolved species produced characteristic electrospray ions, while others of lower relative mobility did not. The TOF mass spectra of cytochrome c, injected from the ion mobility spectrometer at an indicated temperature of 90 degrees C or lower, showed signs that were characteristic of protein-solvent clustering. PMID- 21639213 TI - Oligosaccharide dehydrogenase-modified graphite electrodes for the amperometric determination of sugars in a flow injection system. AB - Enzyme electrodes for the determination of sugars based on solid graphite electrodes modified with oligosaccharide dehydrogenase "wired" with an osmium based one-electron (no proton) acceptor redox hydrogel were studied as sensors in a flow injection system. The enzyme and a poly(1-vinylimidazole) (PVI) where every tenth mer is complexed with osmium (4,4'-dimethylbpy)(2)Cl, (denoted PVI(10)dmeOs) were cross-linked with poly(ethylene glycol) (diglycidyl) ether. The electrodes were active for l-arabinose, d-xylose, d-galactose, d-fructose, d glucose, d-mannose, cellobiose, lactose, maltose, and maltooligosaccharides up to a degree of polymerization of 7. The highest relative response found was for glucose (100%) followed by maltose (40.6%) and lactose (40.6%). Fructose and isomaltotriose gave the lowest responses (<1%). Calibration curves for glucose were strictly linear in the range between 30 and 500 MUM with sensitivity and apparent Michaelis-Menten constant (K(m)(app)) of 23.0 +/- 1.4 MUA mM(-)(1)cm( )(2) and 4.26 +/- 0.95 mM, respectively. PMID- 21639214 TI - Electrochemical determination of water in environmental hydraulic fluids using the karl Fischer reaction. AB - Different procedures based on the Karl Fischer reaction were investigated with respect to their applicability for water determinations in environmental hydraulic fluids: (i) continuous coulometry using a recently described diaphragm free cell; (ii) on-line stripping of water at elevated temperature using either continuous coulometry or direct potentiometry for detection of the liberated water. Except for one of the oils, Statoil PA, which is a poly(alpha-olefin) with certain polymers added, no significant difference was found among coulometry using an optimized imidazole-buffered methanolic reagent containing 75% (v/v) chloroform, the two different stripping techniques (working in the temperature interval 100-110 degrees C), and the commercially available Hydranal Coulomat AG H. The high stability and sensitivity of the coulometric technique described made it possible to work with sample amounts in the low milligram-range, and this is shown to increase the reliability of the coulometric method as compared to normally used procedures. PMID- 21639215 TI - Electrochemical measurements of oligonucleotides in the presence of chromosomal DNA using membrane-covered carbon electrodes. AB - Substantial improvements in the selectivity of electrochemical measurements of trace nucleic acids are obtained by using membrane-covered carbon disk electrodes. Access to the electrode surface can be manipulated via a judicious choice of the membrane molecular weight cutoff (MWCO). The resulting separation step, performed in situ at the electrode surface, adds a new dimension of selectivity based on molecular size to electroanalysis of nucleic acids. Transport properties are evaluated with respect to the oligonucleotide length and membrane MWCO. A highly selective response is observed for synthetic oligonucleotides in the presence of otherwise interfering chromosomal DNAs. Discrimination among oligonucleotides of different lengths is also possible. Short accumulation periods (1-5 min) are sufficient for convenient measurements of low milligram per liter concentrations. PMID- 21639216 TI - Modeling potentiometric sensitivity of conducting polymers. AB - The influence of different polymerization conditions and electrochemical processes provoked by soaking on the potentiometric sensitivity of conducting polymer films is discussed. Poly(pyrrole) doped with hexacyanoferrate(II) is selected as a model polymer. It is shown that, depending on the conditions applied during film deposition and soaking, either anionic or cationic potentiometric responses can be observed and related to the composition of the film. The potentiometric sensitivity of the conducting polymer films is analyzed and interpreted by means of generalized theoretical schema. PMID- 21639217 TI - Mechanistic Studies of the Electrocatalytic Oxidation of NADH and Ascorbate at Glassy Carbon Electrodes Modified with Electrodeposited Films Derived from 3,4 Dihydroxybenzaldehyde. AB - Studies of the electrocatalytic oxidation of beta-nicotinamide adenine dinucleotide (NADH) at glassy carbon rotated disk electrodes modified with electrodeposited films derived from 3,4-dihydroxybenzaldehyde (3,4-DHB) indicate that the mechanism of such electrooxidation proceeds via the formation of an intermediate complex. The reaction also appears to be strongly influenced by the presence of Ca(2+) and Mg(2+) ions as well as by pH. Ascorbate can also be electrocatalytically oxidized at these modified electrodes, giving rise to an electrochemical response very similar to that obtained for NADH. Due to this similarity, the presence of ascorbate in NADH determinations presents a severe interference that cannot be mitigated on the basis of electrochemical responses alone. However, this interference effect can be virtually suppressed by the presence of ascorbate oxidase in solution or immobilized on a nylon mesh which, in turn, is in contact with the electrode modified with the film of 3,4-DHB. Using this approach, we describe the construction of an alcohol biosensor based on alcohol dehydrogenase and which is, furthermore, free from interference effects due to ascorbate. PMID- 21639218 TI - Unnatural isotopic composition of lithium reagents. AB - Isotopic analysis of 39 lithium reagents from several manufacturers indicates that seven were artificially depleted in (6)Li significantly in excess of the variation found in terrestrial materials. The atomic weight of lithium in analyzed reagents ranged from 6.939 to 6.996, and delta(7)Li, reported relative to L-SVEC lithium carbonate, ranged from -11 to +30130/00. This investigation indicates that (6)Li-depleted reagents are now found on chemists' shelves, and the labels of these (6)Li-depleted reagents do not accurately reflect the atomic and (or) molecular weights of these reagents. In 1993, IUPAC issued the following statement: "Commercially available Li materials have atomic weights that range between 6.94 and 6.99; if a more accurate value is required, it must be determined for the specific material." This statement has been found to be incorrect. In two of the 39 samples analyzed, the atomic weight of Li was in excess of 6.99. PMID- 21639219 TI - Chromatography on water-ice. AB - Ice contains a thin film of water on its surface, the thickness ranging from nanometer to micrometer levels, depending on the bulk temperature and the composition of the solution used in making the ice. It follows that water held on the surface of an ice particle should endow a column packed with microparticulate ice the properties of a chromatographic column containing a pellicular packing. We demonstrate here the normal phase separation of two dyes on a column packed with millimeter-size ice particles, held at -18 degrees C. PMID- 21639220 TI - Characterization by potentiometric procedures of the Acid-base and metal binding properties of two new classes of immobilized metal ion affinity adsorbents developed for protein purification. AB - The acid-base protonation constants of two recently introduced chelating ligands for protein purification, O-phosphoserine and 8-hydroxyquinoline immobilized onto Sepharose CL-4B, and the stability constants of their derived immobilized metal ion chelate complexes have been determined by potentiometric methods. The data confirm that immobilization thermodynamically constrains the ligands, with the electron-withdrawing characteristics of the group linking the ligand to the support material affecting the magnitude of the stability constant of the immobilized metal ion complex vis-a-vis the free ligand-metal ion complex in solution. The influence of buffer composition, ionic strength, and pH on the stability constant of the immobilized hard metal ion chelate complexes has also been examined. Collectively, the results have confirmed that coordination complexes with stoichiometries other than the simply 1:1 ML-type exist with these systems, with hard metal ions exhibiting a preference for hydrolytic M(OH)(m)L(n) complexes where m or n > 1. These findings on the participation of coordination complexes of different stoichiometry depending on the characteristics of the chelating ligand and metal ion have fundamental implications for the interpretation of immobilized metal ion affinity chromatographic separation of proteins. PMID- 21639221 TI - Determination of Chlorinated Hydrocarbons in Marine Sediments at the Part-per Trillion Level with Supercritical Fluid Extraction. AB - An off-line supercritical fluid extraction method has been developed for the determination of polychlorinated biphenyls and organochlorine pesticides at the part-per-trillion level in marine sediments. Four different extraction conditions were evaluated: pure carbon dioxide at 60 degrees C/220 atm and 100 degrees C/350 atm; addition of methanol prior to the static extraction at 80 degrees C/350 atm; addition of methanol prior to the static extraction step and continuously during the dynamic extraction step at 80 degrees C/350 atm. Only the latter condition gave extraction recoveries that were comparable and for some compounds higher than Soxhlet recoveries. The significance of differences between the two extraction methodes was determined in an F- and t-test. The method was validated by analysis of a reference material from an international intercomparison exercise. PMID- 21639222 TI - Pervaporation as an alternative to headspace. AB - The use of pervaporation as an alternative to headspace is proposed. The analytical system involves the speciation of organomercury compounds in solid samples using pervaporation, which has been coupled for the first time to gas chromatography. The speciation of mercury as Me(2)Hg, Et(2)Hg, and MeHgCl has been carried out without any derivatization of the analytes, which, after separation from the solid matrix, are preconcentrated on a Tenax minicolumn prior to desorption and chromatographic separation on a semicapillary column (HP-1) prior to atomic fluorescence detection. No column degradation was observed. Linear ranges and detection limits slightly better than those obtained by headspace GC were observed for mercury species in solid samples. Excellent recoveries (between 95 and 107%) for mercury species added to complex solid samples were obtained by this extremely simple and easily automated setup. PMID- 21639223 TI - Potentiometric properties of ion-selective electrode membranes based on segmented polyether urethane matrices. AB - Potentiometric responses of polyurethane (PU)-based membranes containing valinomycin and varying amounts of plasticizer (DOA) and/or lipophilic additive (KTpClPB) were examined as a function of soft segment [poly(tetramethylene ether glycol)] contents in aromatic diisocyanate-based PU matrices. Upon increasing the weight percentages (w(soft)) of soft segments, which in part behave like a built in plasticizer, providing the matrices with rubbery structure (glass transition temperature below -58 degrees C), the amounts of DOA and/or KTpClPB necessary to result in near-Nerntian response (e.g., slope > 50 mV/decade) to potassium were substantially lowered. The apparent effect of adding plasticizer to PU-based membranes was comparable to that resulting from an increase of free carrier concentration in normal PVC-based membranes. Owing to the chemical interaction between mobile anionic sites and urethane chains, plasticizer-free PU membranes could be prepared with the PU matrices with high soft segment contents (w(soft) >= 60 wt %). PUs composed of 60 <= w(soft) < 80 wt % were recommended as the matrix for fabricating ISE membranes with no or low plasticizer content. PMID- 21639224 TI - Determination of cobalt in seawater by catalytic adsorptive cathodic stripping voltammetry. AB - A new procedure for the direct determination of picomolar levels of cobalt in seawater is presented. Cathodic stripping voltammetry is preceded by adsorptive accumulation of the cobalt-nioxime (cyclohexane-1,2-dione dioxime) complex from seawater containing 6 MUM nioxime and 80 mM ammonia at pH 9.1, onto a hanging mercury drop electrode, followed by reduction of the adsorbed species. The reduction current is catalytically enhanced by the presence of 0.5 M nitrite. Optimized conditions for cobalt include a 30 s adsorption period at -0.7 V and a voltammetric scan using differential pulse modulation. According to the proposed reaction mechanism, dissolved Co(II) is oxidized to Co(III) upon addition of nioxime and high concentrations of ammonia and nitrite; a mixed Co(III)-ammonia nitrite complex is adsorbed on the electrode surface; the Co(III) is reduced to Co(II) (complexed by nioxime) during the voltammetric scan, followed by its chemical reoxidation by the nitrite, initiating a catalytically enhanced current. A detection limit of 3 pM cobalt (at an adsorption period of 60 s) enables the detection of this metal in uncontaminated seawater using a very short adsorption time. UV digestion of seawater is essential, as part of the cobalt may occur strongly complexed by organic matter and rendered nonlabile. The method was applied successfully to the determination of the distribution of cobalt in the water column of the Mediterranean. PMID- 21639225 TI - Reagentless mediated laccase electrode for the detection of enzyme modulators. AB - We have investigated aerobic mediation of electron transfer to a laccase enzyme by the solution redox couples [Os(bpy)(2)Cl(2)](+/0) and [Os(bpy)(2)(MeIm)Cl](2+/+), where bpy is 2,2-bipyridine and MeIm is N methylimidazole. The factors that influence the homogeneous mediation reaction are investigated and discussed. Investigation of ionic strength, pH, and temperature effects on the kinetics of intermolecular electron transfer elucidates the governing factors in the mediator-enzyme reactions. Coimmobilization of both enzyme and an osmium redox mediator in a hydrogel on glassy carbon electrodes results in a biosensor for the reagentless addressing of enzyme activity, consuming only oxygen present in solution. Thus, these immobilized enzyme biosensors can be utilized for the detection of modulators of laccase activity, such as the inhibitor sodium azide. The enzyme inhibition biosensor can detect levels of azide as low as 2.5 * 10(-6) mol dm(-3) in solution. PMID- 21639226 TI - Electrochemical/Piezoelectric dual-response biosensor for heme ligands. AB - A heme peptide-modified quartz crystal oscillator is fabricated as a dual response biosensor, giving electrochemical and piezoelectric responses simultaneously. Binding of a ligand to heme peptide causes inhibition of its catalytic activity, which is observed as the electrochemical response, and a mass increase is monitored as the piezoelectric response. The dual-response sensor can give two independent pieces of information at the same time. That is, qualitative (identification of the species) and quantitative (determination of its concentration) analyses can be made simultaneously. Simultaneous quantitative analysis for two components is also possible. These two modes are theoretically explained and then experimentally demonstrated employing imidazole and histidine as interferants. PMID- 21639227 TI - Selective Determination of Cr(VI) by a Self-Assembled Monolayer-Based Electrode. AB - We have developed a selective electrode for chromium(VI), based on a self assembled monolayer of 4-(mercapto-n-alkyl)pyridinium on gold surfaces, which exhibits unique speciation capabilities. Cr(VI) levels as low as 1 parts per trillion can be detected using a 4-(mercaptoethyl)pyridinium monolayer. The different parameters that govern the analytical performance of these electrodes have been studied in detail and optimized. In addition, the organization of the monolayers has been examined by a variety of surface techniques such as XPS, FT IR, and electrochemistry. Our results show that structuring and understanding the solid-liquid interface at the molecular level are essential for designing probes with superior analytical characteristics. PMID- 21639228 TI - Electrochemical behavior and determination of the insecticide synergist piperonyl butoxide. AB - Piperonyl butoxide may be reversibly oxidized in acetonitrile at a glassy carbon electrode to a cation radical under short time scale voltammetric conditions, e.g., cyclic voltammetry when the potential scan rate is above 500 mV s(-)(1). During longer time domain experiments, the cation radical decays in a rate limiting heterolytic bond cleavage step and subsequent transfer of a second electron at the potential of the first process. Additionally, a second oxidation process develops at more positive potentials. One product isolated from the initial oxidation process in an almost quantitative yield, under controlled potential electrolysis conditions, is 6-n-propyl-1,3-benzodioxole-5 carboxaldehyde. This carboxaldehyde is oxidized at the same positive applied potential as the second oxidation process observed in long time domain voltammetric experiments with piperonyl butoxide. The limit of detection for piperonyl butoxide in acetonitrile, using differential pulse voltammetry at a glassy carbon electrode, is 1.6 * 10(-)(6) M (3sigma), with a limit of determination of 4.1 * 10(-)(6) M (10sigma). Piperonyl butoxide was selectively determined using differential pulse voltammetry with a concentration of 5.11 +/- 0.02 g L(-)(1) in a commercial insecticide formulation containing pyrethrins. This result is in good agreement with the manufacturer's stated concentration of 5.07 g L(-)(1). The sample preparation requires only simple dilution of the formulation in an acetonitrile/dichloromethane (95:5) solvent mixture. PMID- 21639229 TI - Stable nitrogen isotope analysis of amino Acid enantiomers by gas chromatography/combustion/isotope ratio mass spectrometry. AB - The analysis of the stable nitrogen isotope compositions of individual amino acid stereoisomers through the use of gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) is presented. Nitrogen isotopic compositions of single amino acids or of their enantiomers is possible without the labor-intensive and time-consuming preparative-scale chromatographic procedures required for conventional stable isotope analysis. Following hydrolysis and derivatization, single-component isotope analysis is accomplished on nanomole quantities of each of the stereoisomers of an amino acid, utilizing the effluent stream of gas chromatographic separation. Nitrogen isotope fractionation is minimal during acylation of the amino acid, with no additional nitrogen being added stoichiometrically to the derivative. Thus, the isotopic composition of the nitrogen in the derivative is that of the original compound. Replicate stable nitrogen isotope analyses of 11 amino acids, and their trifluoroacetyl (TFA)/isopropyl (IP) ester derivatives, determined by both conventional isotope ratio mass spectrometry (IRMS) and GC/C/IRMS, indicate that the GC procedure is highly reproducible (standard deviations typically 0.3-0.40/00) and that isotopic differences between the amino acid and its TFA/IP derivative are, in general, less than 0.50/00. PMID- 21639230 TI - Theoretical and experimental studies of the effect of pressure on solute retention in liquid chromatography. AB - Pressure is often assumed to have a negligible influence on solute retention in liquid chromatography because of the small compressibility of the mobile and stationary phases. The range of pressures commonly encountered in reversed-phase separations is considerable, however, and may give rise to significant changes in solute capacity factor. In this study, the retention of model solutes is measured directly along the chromatographic column as a function of the local pressure. The model solutes, a homologous series of derivatized fatty acids, exhibit a significant increase in capacity factor ranging from +9.3% for n-C(10) to +24.4% for n-C(20) for inlet pressures from 1500 to 5000 psi. These experimental results are compared with a thermodynamic model derived from regular solution theory. This model suggests that state effects alone are not sufficient to describe the measured change in solute retention and that variations in interaction energy with density must also be considered. By using the simple relationship of van der Waals for the interaction energy (E ? 1/V), the change in capacity factor with density is slightly underestimated. However, by using an extended relationship that better describes polar fluids (E ? 1/V(2)), good agreement is observed. Finally, the correlation of experimental results with this thermodynamic model reveals that all components in the chromatographic system, including the solute, mobile phase, and stationary phase, must be considered compressible. The results of this study have clear implications for the determination of fundamental physicochemical parameters, as well as for the everyday practice of liquid chromatography. PMID- 21639231 TI - Carbon Isotope Analyses of Semivolatile Organic Compounds in Aqueous Media Using Solid-Phase Microextraction and Isotope Ratio Monitoring GC/MS. AB - Solid-phase microextraction (SPME) was used to facilitate the measurement of stable carbon isotope compositions (at natural abundance) of six organic compounds representing four compound classes in aqueous solution. Toluene, methylcyclohexane, hexanol, and acetic, propionic, and valeric acids were extracted from aqueous solutions with appropriate SPME phases and thermally desorbed into the split/splitless inlet of an isotope ratio monitoring gas chromatograph/mass spectrometer (irmGC/MS). Hydrophobic compounds (toluene, methylcyclohexane, hexanol) extracted by a nonpolar SPME phase were slightly (<=0.50/00) enriched in (13)C while organic acids extracted with a polar phase were depleted in (13)C to a somewhat greater degree (<=1.50/00) relative to material remaining in the aqueous phase. Isotopic fractionation was not observed to vary systematically as a function of equilibration time or solute concentration. Further, isotope fractionation did not vary consistently with the partition coefficient (K(fw)). However, both salinity and cosolvent effects, which altered the partition coefficients of the solutes, also yielded a reduction in the magnitude of isotopic fractionation (to <=0.40/00 for the hydrocarbons, <=0.50/00 for the organic acids). We conclude that fractionations are most likely associated with the interactions of organic compounds with the organic phase coating SPME fibers and are specifically due to mass-dependent energy shifts upon solution of each analyte into the organic phase. In addition, fractionations are also influenced by energy shifts associated with electrostatic forces acting on the analyte in the water phase during the partitioning process. The magnitude of isotopic fractionations can be minimized under conditions appropriate for the analysis of natural waters, and with careful calibration, SPME and irmGC/MS should be a valuable means for isotopic analyses for a wide range of organic constituents in aqueous samples. PMID- 21639232 TI - Integrated Capillary Electrophoresis/Electrochemical Detection with Metal Film Electrodes Directly Deposited onto the Capillary Tip. AB - The practical application of electrochemical detection in capillary electrophoresis has been hampered by irreproducibility and inconvenience related to capillary/electrode alignment. In order to eliminate these problems, a simple, flexible method by which the capillary and the working electrode were integrated into a single operational unit was devised and evaluated. The electrodes were formed by sputtering a thin conductive layer of Au or Pt onto the exit tip of the capillary. Depending on the size of the capillary used (i.e., both inner and outer diameters), Au on-capillary electrodes (OCEs) gave detection limits at the micromolar level and slightly below for the test analytes dopamine and catechol. More important, operation of the OCEs required no alignment procedures beyond immersion in the CE buffer reservoir/detector cell. OCEs used in this manner exhibited relative standard deviations of 2-4% for repeated injections even if removed from solution between runs. Finally, the Au and Pt OCEs could themselves be modified further by conventional electrochemical procedures. Here, Cu OCEs, formed by electrodeposition onto Au, were used to detect carbohydrate compounds; and an enzyme OCE, formed by adsorption of glucose oxidase onto Pt, was used to detect glucose. PMID- 21639233 TI - Optimization of SFE for the Determination of Chlorinated Hydrocarbons at the Part per-Trillion Level. AB - The applicability of SFE for trace analysis was expanded by an intensive cleaning procedure of the SFE equipment and development of an adsorbing device for commercial carbon dioxide. For a selection of chlorinated hydrocarbons, the limit of detection was reduced to <=20 pg/g of solid matrix. The procedure can be transferred to other analytical SF delivery systems for off- and on-line SFE, and also to an SFC apparatus. Also, the blank value for the analysis of nonchlorinated aliphatic and aromatic compounds was improved significantly. The purity of carbon dioxide is no longer the limiting factor for trace analysis with SFE. PMID- 21639234 TI - An electrochemical method for making enzyme microsensors. Application to the detection of dopamine and glutamate. AB - A novel method of microbiosensor fabrication is described. It is based on the electrochemical polymerization of an enzyme-amphiphilic pyrroleammonium solution on the surface of a microelectrode in the absence of supporting electrolyte. By trapping glutamate oxidase (GMO) or polyphenol oxidase (PPO) in such polypyrrole films, we made microbiosensors for the amperometric determination of glutamate or dopamine, respectively. The response of the GMO microelectrode to glutamate was based on the amperometric detection of the enzymically generated hydrogen peroxide at 0.6 V vs SCE. The detection limit and sensitivity of this microbiosensor were 1 MUM and 32 mA M(-1) cm(-2), respectively. The response of the PPO microelectrode to dopamine was based on the amperometric detection of the enzymically generated quinoid product at -0.2 V. The calibration range for dopamine measurement was 5 * 10(-8)-8 * 10(-5) M and the detection limit and sensitivity were 5 * 10(-8) M and 59 mA M(-1) cm(-2), respectively. PMID- 21639235 TI - Evaluation of a mirror-polishing technique for fluorocarbon polymer surfaces for reduction of contamination from containers used in ultratrace analysis. AB - A mirror-polishing technique for fluorocarbon polymer surfaces using high precision diamond cutting tools was developed. The goal of this technique was the reduction of ultratrace elemental analysis contamination levels of containers fabricated from such mirror-polished materials. Remarkably smooth inner surfaces with degrees of flatness of 0.1 MUm peak-to-valley (PTV) for containers fabricated from mirror-polished PTFE materials were obtained, in contrast to degrees of surface flatness of more than 30 MUm PTV for commercially available PTFE containers. (Here, PTV denotes the difference between the highest peak and deepest valley in a scanned area of 10 * 10 MUm.) Extractable impurity levels for mirror-polished PTFE container surfaces were reduced by more than 1 order of magnitude relative to those of unpolished PTFE containers. The surface conditions of the PTFE containers were observed by atomic force and scanning electron microscopy. The microphotographs so obtained suggest that the degree of surface smoothness of the containers is proportional to their ultratrace metallic contamination levels. PMID- 21639236 TI - Real-Time Imaging of Microscopic pH Distribution with a Two-Dimensional pH Imaging Apparatus. AB - Real-time two-dimensional pH imaging has become possible with the development of a new type of potentiometry, by which the pH distribution can be visualized as pH images with submillimeter spatial resolution. The measurement time for one image is short enough for practical use. The transient change of pH distribution generated by the protons released from a single cation-exchange resin was observed and visualized as the pH images. From the pH images, the evaluation of the resin performance was also carried out. PMID- 21639237 TI - Modulated FT-Raman Fiber-Optic Spectroscopy: A Technique for Remotely Monitoring High-Temperature Reactions in Real-Time. AB - A modification to a commercial FT-Raman spectrometer is presented for the elimination of thermal backgrounds in FT-Raman spectra. The modification involves the use of a mechanical chopper to modulate the CW laser, remote collection of the signal via fiber optics, and connection of a dual-phase digital signal processor lock-in amplifier between the detector and the spectrometer's collection electronics to demodulate and filter the optical signals. The resulting modulated FT-Raman fiber-optic spectrometer is capable of completely eliminating thermal backgrounds at temperatures exceeding 370 degrees C. In addition, the signal/noise of generated Raman spectra is greater than for spectra collected with the conventional FT-Raman under identical conditions and incident laser power. This is true for both room-temperature and hot samples. The method allows collection of data using preexisting spectrometer software. The total cost of the modification (excluding fiber optics) is ~$3000 and requires less than 2 h to implement. This is the first report of FT-Raman spectra collected at temperatures in excess of 300 degrees C in the absence of thermal backgrounds. PMID- 21639238 TI - Water analysis by solid phase microextraction based on physical chemical properties of the coating. AB - A novel and simple method using solid phase microextraction (SPME) with poly(dimethylsiloxane) fiber coating and linear temperature-programmed retention index (LTPRI) has been developed to quantify petroleum hydrocarbons in water. Partition coefficients (K(fw)) for the analytes in water and SPME were established for a series of aromatics, cycloalkanes and alkanes. There is a linear relationship between log K(fw) for each hydrocarbon series and LTPRI. The slope of the curves for all the series are the same and related to the partial free energy of solution for the hydrocarbon-fiber coating solution. The y intercept of the plots is distinct for each group of hydrocarbons and related to Henry's law coefficients for each series. Therefore, the K(fw) for a series of hydrocarbons can be estimated using literature Henry's law coefficients. This method was used to quantify BTEX-contaminated water. PMID- 21639239 TI - Mechanistic aspects of chiral discrimination on modified cellulose. AB - Cellulose and cellulose derivatives are biopolymers which are often used as stationary phases for the separation of enantiomers. Describing the mechanism of such separations is a difficult task due to the complexity of these phases. In the present study, we attempt to elucidate the types of interactions occurring between a diol intermediate for a LTD(4) antagonist and a tris(4-methylbenzoate) derivatized cellulose stationary phase. Thermodynamic studies indicate that, at low temperatures, the enantioselectivity is entropy driven. At higher temperatures, the separation is enthalpy driven. DSC and IR experiments reveal that the transitions between the enthalpic and the entropic regions of the van't Hoff plots are a result of a change in conformation of the stationary phase. Investigation of chromatographic kinetic parameters reveals that, at low temperature, the second eluted enantiomer undergoes sluggish inclusion interactions. Subtle changes in the structure of the analyte indicates that pi-pi interactions do not contribute to enantioselectivity. Finally, molecular modeling of (R)- and (S)-diol and the stationary phase suggests that hydrogen bonding is a primary factor in the separation, and the calculated energy values obtained from the molecular modeling correlate well with the chromatographic elution order. PMID- 21639240 TI - Optical detection of chloramphenicol using molecularly imprinted polymers. AB - A practical optical sensing system for the determination of chloramphenicol (CAP), utilizing molecularly imprinted polymers (MIPs) and HPLC, has been developed. The method is based on competitive displacement of a chloramphenicol methyl red (CAP-MR) dye conjugate from specific binding cavities in an imprinted polymer by the analyte. The best of these polymers was obtained using (diethylamino)ethyl methacrylate as functional monomer at a monomer:template ratio of 2:1. HPLC with a mobile phase containing CAP-MR was used as the detection system, and injection of CAP and, to a lesser degree, thiamphenicol resulted in proportional displacement of the conjugate, which was detected at 460 nm. The detection system showed a linear response over a range of 3-1000 MUg/mL and effectively detected CAP extracted from serum. This system offers a tailor made, selective, and rapid method for CAP detection, is able to discriminate between similar molecules, and is effective below and above the therapeutic range (10-20 MUg/mL serum, potentially toxic above 25 MUg/mL). This technique is quite general and should enable the use of MIPs in a wide variety of applications involving the detection of families of molecules which possess a distinct arrangement of functional groups. PMID- 21639241 TI - Chromatographic Hydrophobicity Index by Fast-Gradient RP-HPLC: A High-Throughput Alternative to log P/log D. AB - A new chromatographic hydrophobicity index (CHI) is described which can be used as part of a protocol for high-throughput (50-100 compounds/day) physicochemical property profiling for rational drug design. The index is derived from retention times (t(R)) observed in a fast gradient reversed-phase HPLC method. The isocratic retention factors (log k') were measured for a series of 76 structurally unrelated compounds by using various concentrations of acetonitrile in the mobile phase. By plotting the log k' as a function of the acetonitrile concentration, the slope (S) and the intercept (log k'(w)) values were calculated. The previously validated index of hydrophobicity phi(0) was calculated as -log k'(w)/S. A good linear correlation was obtained between the gradient retention time values, t(R) and the isocratically determined phi(0) values for the 76 compounds. The constants of this linear correlation can be used to calculate CHI. For most compounds, CHI is between 0 and 100 and in this range it approximates to the percentage (by volume) of acetonitrile required to achieve an equal distribution of compound between the mobile and the stationary phases. CHI values can be measured using acidic, neutral, or slightly basic eluents. Values corresponding to the neutral form of molecules could be measured for 52 of the compounds and showed good correlation (r = 0.851) to the calculated octanol/water partition coefficient (c log P) values. PMID- 21639242 TI - A gas chromatographic separation for the h and C stable isotope ratio determination of coal compounds. AB - A new, completely automated gas chromatography technique has been developed to separate the different gaseous compounds produced during underground coal gasification for their (13)C/(12)C and D/H isotope ratio measurements. The technique was designed for separation and collection of H(2), CO, CO(2), H(2)O, H(2)S, CH(4), and heavier hydrocarbons. These gaseous compounds are perfectly separated by the gas-phase chromatograph and quantitatively sent to seven combustion and collection lines. H(2), CO, CH(4), and heavier hydrocarbons are quantitatively oxidized to CO(2) and/or H(2)O. The isotopic analyses are performed by the sealed-tube method. The zinc method is used for reduction of both water and H(2)S to hydrogen for D/H analysis. Including all preparation steps, the reproducibility of isotope abundance values, for a quantity higher than or equal to 0.1 mL of individual components in a mixture (5 mL of gases being initially injected in the gas chromatograph), is +/-0.10/00 for delta(13)C(PDB) and +/-60/00 for deltaD(SMOW). PMID- 21639243 TI - UV Laser Machined Polymer Substrates for the Development of Microdiagnostic Systems. AB - This report describes a UV laser photoablation method for the production of miniaturized liquid-handling systems on polymer substrate chips. The fabrication of fluid channel and reservoir networks is accomplished by firing 200 mJ pulses from an UV excimer laser at substrates moving in predefined computer-controlled patterns. This method was used for producing channels in polystyrene, polycarbonate, cellulose acetate, and poly(ethylene terephthalate). Efficient sealing of the resulting photoablated polymer channels was accomplished using a low-cost film lamination technique. After fabrication, the ablated structures were observed to be well defined, i.e., possessing high aspect ratios, as seen by light, scanning electron, and atomic force microscopy. Relative to the original polymer samples, photoablated surfaces showed an increase in their hydrophilicity and rugosity as a group, yet differences were noted between the polymers studied. These surface characteristics demonstrate the capability of generating electroosmotic flow in the cathodic direction, which is characterized here as a function of applied electric field, pH, and ionic strength of common electrophoretic buffer systems. These results show a correlation between the ablative changes in surface conditions and the resulting electroosmotic flow. The effect of protein coatings on ablated surfaces is also demonstrated to significantly dampen the electroosmotic flow for all polymers. All of these results are discussed in terms of developing liquid-handling capability, which is an essential part of many MU-TAS and chemical diagnostic systems. PMID- 21639244 TI - Quartz crystal microbalance study of DNA immobilization and hybridization for nucleic Acid sensor development. AB - The immobilization of two 30-mer oligonucleotides, one biotinylated (biotin-DNA) and the other having a mercaptohexyl group at the 5'-phosphate end (BS1-SH), onto modified gold surfaces has been examined using a quartz crystal microbalance (QCM). Both single-layer and multilayer DNA films were prepared. The single-layer films of biotin-DNA were constructed by binding to a precursor layer of avidin, which had been attached to the QCM either covalently using a water-soluble carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) or via electrostatic interaction with poly(allylamine hydrochloride) (PAH). Single-layer films of BS1-SH were also formed on PAH via the electrostatic attraction between the amine groups on PAH and the negatively charged phosphate backbone of DNA. Multilayer films of DNA were fabricated by the successive deposition of avidin and poly(styrenesulfonate) (PSS), up to a total of nine avidin/PSS layers, followed by DNA adsorption. DNA immobilization and hybridization of the immobilized DNAs was monitored in situ from QCM frequency changes. Hybridization was induced by exposure of the DNA-containing films to complementary DNA in solution. Equal frequency changes were observed for the DNA immobilization and hybridization steps for the single-layer films, indicating a DNA probe-to hybridized DNA target ratio of 1:1. The multilayer DNA films also exhibited DNA hybridization, with a greater quantity of DNA hybridized compared with the single layer films. The multilayer films provide a novel means for the fabrication of DNA-based thin films with increased capacity for nucleic acid detection. PMID- 21639245 TI - Resonator/Oscillator response to liquid loading. AB - The resonant frequency of a thickness-shear mode resonator operated in contact with a fluid was measured with a network analyzer and with an oscillator circuit. The network analyzer measures changes in the device's intrinsic resonant frequency, which varies linearly with (rhoeta)(1/2), where rho and eta are liquid density and viscosity, respectively. The resonator/oscillator combination, however, responds differently to liquid loading than the resonator alone. By applying the operating constraints of the oscillator to an equivalent-circuit model for the liquid-loaded resonator, the response of the resonator/oscillator pair can be determined. By properly tuning the resonator/oscillator pair, the dynamic range of the response can be extended and made more linear, closely tracking the response of the resonator alone. This allows the system to measure higher viscosity and higher density liquids with greater accuracy. PMID- 21639246 TI - Determination of heterogeneous electron transfer kinetics in the presence of ultrasound at microelectrodes employing sampled voltammetry. AB - The technique of sampled voltammetry at microelectrodes irradiated with ultrasound is demonstrated for the first time. This technique is used to determine the heterogeneous electron transfer rate constants for the redox couples ferrocene/ferrocenium, Ru(NH(3))(6)(3+/2+), and IrCl(6)(3)(-)/IrCl(6)(2)( ). Determination of the heterogeneous rate constants is also achieved for comparison purposes by analysis of fast sweep rate voltammetry of the redox systems studied at microelectrodes and comparison of the results obtained to the theory developed by Nicholson and Shain. The heterogeneous rate constants determined using sampled voltammetry were 1.0, 0.6, 1.23, and 0.18 cm s(-)(1) for the ferrocene/ferrocenium (0.1 mol dm(-)(3) TEATFB, CH(3)CN), Ru(NH(3))(6)(3+/2+) (0.1 mol dm(-)(3) KCl), IrCl(6)(3)(-) (1 mol dm(-)(3) KCl), and IrCl(6)(3)( )/IrCl(6)(2)(-) (1 mol dm(-)(3) NaCl), respectively, in agreement with those obtained in the absence of ultrasound. PMID- 21639247 TI - Determination of the diffusion coefficient of hydrogen in aqueous solution using single and double potential step chronoamperometry at a disk ultramicroelectrode. AB - An assessment is made of single and double potential step chronoamperometry (SPSC and DPSC, respectively) at Pt disk ultramicroelectrodes (UMEs) as methods for determining the value of the diffusion coefficient of hydrogen in aqueous solutions. In SPSC, measured currents for the oxidation of dissolved hydrogen (at concentrations close to saturated solution values) comprise a significant contribution, at short to moderate times, from the oxidative desorption of adsorbed hydrogen as well as the diffusion-controlled oxidation of the solution species. Provided that the electrode is preconditioned using a well-defined potential cycling procedure, the behavior for the oxidative desorption step alone can be established in an Ar-saturated solution. The chronoamperometric characteristics for the solution diffusion-controlled process may then be determined, from which the diffusion coefficient of hydrogen can be measured. In DPSC, a locally supersaturated solution of hydrogen is created transiently through the diffusion-controlled reduction of a known concentration of protons in an initial potential step. Hydrogen is subsequently collected back through oxidation to protons; the current flowing depends on the diffusion coefficients of the two species and the duration of the forward step. Under these conditions, the contribution from surface electrochemical processes to the forward and reverse chronoamperommograms is shown to be negligible. By solving the mass transport problem for DPSC with arbitrary diffusion coefficients of the redox species, the diffusion coefficient of hydrogen is readily determined. Both methods yield a consistent value for the diffusion coefficient of hydrogen, D(H)((2)), in 0.1 mol dm(-)(3) KNO(3) of 5.0 * 10(-)(5) cm(2) s(-)(1). PMID- 21639248 TI - Analysis of the magnetic force generated at a hemispherical microelectrode. AB - An analytical expression is presented for the magnetic force generated during steady-state voltammetry at a hemispherical microelectrode immersed in a uniform magnetic field. Diffusion of electrogenerated ions through the magnetic field results in a magnetic force that induces convective solution flow near the electrode surface. The magnetic force per unit volume,[Formula: see text] (i.e., force density), is shown to decrease as r(-)(2), where r is the distance away from the center of the hemispherical electrode. A consequence of the inverse square dependence of[Formula: see text] on r is that the magnetic force is confined to a microscopic solution volume near the electrode surface (e.g., ~2 * 10(-)(9) L for a 12.5-MUm-radius hemispherical electrode). The net magnetic force acting on the diffusion layer volume,[Formula: see text] , is computed as a function of magnetic field strength and orientation and used in an approximate analysis of experimental data obtained at an inlaid 12.5-MUm-radius Pt microdisk electrode. Enhancements in voltammetric currents are shown to result from magnetic forces as small as 2 * 10(-)(11) N. PMID- 21639249 TI - Semicontinuous Detection of 1,2-Dichloroethane in Water Samples Using Xanthobacter autotrophicus GJ 10 Encapsulated in Chitosan Beads. AB - A semicontinuous microbial assay for the determination of halogenated short-chain hydrocarbons in water samples was developed. The bacterium Xanthobacter autotrophicus GJ 10 forms dehalogenating enzymes, which liberate the halides in 1,2-dichloroethane as halogen ions. Cells of the organism were immobilized in chitosan beads and placed into a tube reactor, whose outlet was connected to a flow-through cell with chloride-selective potentiometric electrodes. Water samples were delivered continuously to the system, and the EMF was recorded. Both the difference in EMF between blank and sample and the velocity of the EMF change were used for calibration. The detection limit for 1,2-dichloroethane was below 0.5 mg/L; the relative standard deviation was <10%. The effects of several parameters like flow rate and cell density were studied in detail. PMID- 21639250 TI - Composite transducers for amperometric biosensors. The glucose sensor. AB - A new concept of a composite transducer for amperometric biosensors based on the use of a solid substance with amphiphilic character (called a solid binding matrix, SBM) is presented. The electrochemical properties of the transducers prepared with five different SBMs and the characteristics and performance of SBM based glucose sensors prepared by three different methods are described. Biosensor stability is evaluated and discussed. The biosensor was used for the determination of glucose in wine, yielding results which were consistent with those obtained with the commercially available Glucose Enzyme Photometric Kit. The average accuracy was 6% for the whole range of analyzed concentrations (0.2 47 g/L) using the same sample dilution in a buffer. PMID- 21639251 TI - Raman spectroscopic determination of the structure and orientation of organic monolayers chemisorbed on carbon electrode surfaces. AB - Azobenzene (AB) and 4-nitroazobenzene (NAB) were covalently bonded to carbon surfaces by electrochemical reduction of their diazonium derivatives. The N(1s) features of XPS spectra of modified surfaces had intensities expected for monolayer coverage. However, the Raman spectra were significantly more intense than expected, implying an increase in scattering cross section upon chemisorption. A likely explanation is resonance enhancement of the carbon/adsorbate chromophore analogous to that reported earlier for dinitrophenylhydrazine (DNPH) chemisorption. Vibrational assignments indicate that the C-C vibration between azobenzene and the carbon surface is in the 1240 1280 cm(-1) region, and this conclusion is supported by spectra obtained from [(13)C]graphite. Observation of depolarization ratios for 4-nitroazobenzene and DNPH on graphite edge plane indicate that NAB is able to rotate about the NAB/carbon C-C bond, while chemisorbed DNPH is not. The partial multiple bond character of the DNPH linkage to graphite is consistent with the observation that the DNPH pi system remains parallel to the graphitic planes. PMID- 21639252 TI - A renewable liquid droplet method for on-line pollution analysis by multi-photon ionization. AB - A multi-photon ionization based fast conductance (MPI-FC) technique was applied to detect combustion byproduct aerosols. These PAH-polluted aerosols were on-line sampled by means of renewable water microdroplets. The environmental cases considered here have involved such common air contaminants as motor car exhaust gas and cigarette smoke. The possibility of obtaining useful calibration curves has been addressed. Two droplet contamination regimes were clearly observed. These have been argued to be associated with either a volume uniform (i.e., a bulk type) or a surface-favored contamination. The latter regime is possible whenever the increasing droplet contamination extends beyond the solubility saturation of the PAH compounds. Detection limits as low as 1 pg were obtained for pyrene-contaminated renewable microdroplets. PMID- 21639253 TI - Absolute analysis of particulate materials by laser-induced breakdown spectroscopy. AB - We have developed a new data acquisition approach followed by a suitable data analysis for Laser-induced breakdown spectroscopy. It provides absolute concentrations of elements in particulate materials (e.g., industrial dusts and soils). In contrast to the known calibration procedures (based on the ratio of spectral lines), which are applicable only when one component is constant, this approach requires no constant constituent and results in absolute (rather than relative) concentrations. Thus, the major drawback of this analytical method, namely, the signals' instability (especially when particulate materials are concerned) is partially solved. Unlike the commonly used integrated data acquisition, we use a sequence of signals from single breakdown events. We compensate for pulse to pulse fluctuations in an intrinsic way, and the final results do not depend on the presence of any constant component. Extended linear calibration curves are obtained, and limits of detection are improved by 1 order of magnitude relative to previous methods applied to the same samples (e.g., detection limit of 10(-12) g of Zn in aerosol samples). The proposed compensation for pulse variations is based on the assumption that they can be described as a multiplicative effect for both the spectral peaks and a component of the baseline. In other words, we assume that the same fluctuation pattern observed in the spectral peaks is present in the baseline as well. This assumption is shown to hold and is utilized in the proposed method. In addition, a proper data filtering process, which eliminates ill-conditioned spectra, is shown to partially compensate for problems due to the nature of analysis of particulate materials. PMID- 21639254 TI - Influence of Imidazole and Bis(trichlorophenyl) Oxalate in the Oxalyldiimidazole Peroxyoxalate Chemiluminescence Reaction. AB - The complex role of imidazole when used as a catalyst in the bis(2,4,6 trichlorophenyl) oxalate (TCPO) peroxyoxalate chemiluminescence (PO-CL) reaction is explained by the transient formation and subsequent degradation of 1,1' oxalyldiimidazole (ODI). When ODI was used directly as PO-CL reagent, the stability was improved by addition of TCPO as an "imidazole sponge", since ODI is rapidly decomposed in the presence of imidazole. In this way, the imidazole catalyzed degradation of ODI was hindered efficiently. The stability of ODI was also influenced by the storage vessel material. Polymeric bottles were found to be more suitable than glass containers. A comparison was made between the traditionally used reagent TCPO/imidazole (mixed on-line for formation of ODI) and the new reagent combination ODI-TCPO (premixed) with respect to sensitivity, noise, and background. PMID- 21639255 TI - High-efficiency molecular counting in solution: single-molecule detection in electrodynamically focused microdroplet streams. AB - We report fluorescence detection of individual rhodamine 6G molecules using a linear quadrupole to focus streams of microdroplets through the waist of a counterpropagating cw Ar(+) laser. Since the terminal velocity scales as the square of the droplet diameter, the droplet-laser interaction time was "tunable" between 5 and 200 ms by using water samples spiked with a small, variable (2-5% v/v) amount of glycerol. Fluorescence bursts from droplets containing single molecules were clearly distinguished from the blanks in real time with an average signal-to-noise ratio of about 10, limited primarily by photobleaching and droplet size fluctuations (<1%). The volume throughput rates associated with this approach (~10 pL/s) are roughly 10(3) higher than those associated with particle levitation techniques, with minimal sacrifice in sensitivity. Total molecular detection efficiencies of about 80% (at >99% confidence) were obtained for 100 and 15 fM rhodamine 6G solutions, in good agreement with detailed theoretical calculations and statistical limitations. PMID- 21639256 TI - Furnace atomization plasma ionization mass spectrometry. AB - Development and initial characterization of FAPES as an ion source for elemental mass spectrometry is presented. The source is configured with an integrated contact cuvette in which a 40 MHz He plasma is sustained at atmospheric pressure. A differentially pumped interface consisting of a stainless steel sampling tube (0.5 mm i.d.) and tandem skimmer cone serves to sample the plasma from the open end of the cuvette. Mass spectra, characterizing plasma species with the furnace at ambient and elevated temperatures, are relatively structureless and show no evidence of He(+), He(2)(+), or carbonaceous ions. Transient ion signals, generated during the atomization of a number of analytes introduced in solution form, reveal that the plasma contains sufficiently energetic species to ionize elements having ionization potentials as high as 10.45 eV (iodine). Significant ionization does not occur in the absence of the plasma, nor in the extraction interface. Using Pb as a test element, acceptable isotopic abundances can be obtained, with an estimated absolute limit of detection of 10 pg (2 ng/mL relative). Future directions are discussed. PMID- 21639257 TI - Spectroscopic determination of pressure-induced shifts in inclusion complexation equilibria. AB - The effect of modest hydrostatic pressure (<350 bar) on condensed-phase equilibrium processes has been largely overlooked, due in large part to the small compressibility of these phases relative to gases or supercritical fluids. Although the bulk properties of condensed phases are not significantly modified by pressure in this modest regime, the solvation processes driving inclusion complexation may be appreciably affected. In this paper, we examine this hypothesis using steady-state fluorescence spectroscopy to determine the pressure dependence of association constants. The widely used host molecule, beta cyclodextrin, provides an incompressible hydrophobic cavity into which structurally analogous fluorescent probes are encapsulated. By comparing the unique pressure dependencies of these equilibria, the importance of local site solvation and rim interactions in influencing the pressure dependence is demonstrated. The structurally analogous complexes chosen for these studies are expected to have similar pressure-dependent behavior based on comparable solvation structures. However, pressure-induced changes in the association constant for these two analogs are quite distinct, with differences in K(c) ranging from clearly pressure dependent (-14%) to pressure independent over 338 bar. Additional solvation perturbations are observed in the pressure dependence of the quantum efficiency for both complexes (-7.3% and -9.4%). Thus, pressure induced perturbation in the fluorescence properties of the complex need not be accompanied by simultaneous changes in the complexation equilibrium. Finally, these pressure-induced changes in complexation selectivity are important for all measurements conducted under variable pressure conditions, including liquid chromatography and process monitoring. PMID- 21639258 TI - Characterization of the microdialysis junction interface for capillary electrophoresis/microelectrospray ionization mass spectrometry. AB - A capillary electrophoresis/electrospray ionization mass spectrometry (CE/ESI-MS) interface, based on an electric circuit across a microdialysis membrane surrounding a short capillary segment closely connected to the separation capillary terminus, is demonstrated to be sensitive, efficient, and rugged. A microspray type ionization emitter produces a stable electrospray at the low flow rates provided by CE and thus avoids both the need for a makeup liquid flow provided by liquid junction or sheath flow interfaces and the subsequent dilution and reduction in sensitivity. Reproducibility studies and comparisons with CE/UV and the CE/sheath flow interface with ESI-MS are presented. Additionally, postrun acidification via the microdialysis junction interface is demonstrated and shown to be capable of denaturing the holomyoglobin protein noncovalent complex while maintaining separation efficiency. PMID- 21639259 TI - Comparison of ASE and SFE with Soxhlet, Sonication, and Methanolic Saponification Extractions for the Determination of Organic Micropollutants in Marine Particulate Matter. AB - The methods of accelerated solvent extraction (ASE) and supercritical fluid extraction (SFE) of polycyclic aromatic hydrocarbons (PAHs), aliphatic hydrocarbons, and chlorinated hydrocarbons from marine samples were investigated. The results of extractions of a certified sediment and four samples of suspended particulate matter (SPM) were compared to classical Soxhlet (SOX), ultrasonication (USE), and methanolic saponification extraction (MSE) methods. The recovery data, including precision and systematic deviations of each method, were evaluated statistically. It was found that recoveries and precision of ASE and SFE compared well with the other methods investigated. Using SFE, the average recoveries of PAHs in three different samples ranged from 96 to 105%, for ASE the recoveries were in the range of 97-108% compared to the reference methods. Compared to the certified values of sediment HS-6, the average recoveries of SFE and ASE were 87 and 88%, most compounds being within the limits of confidence. Also, for alkanes the average recoveries by SFE and ASE were equal to the results obtained by SOX, USE, and MSE. In the case of SFE, the recoveries were in the range 93-115%, and ASE achieved recoveries of 94-107% as compared to the other methods. For ASE and SFE, the influence of water on the extraction efficiency was examined. While the natural water content of the SPM sample (56 wt %) led to insufficient recoveries in ASE and SFE, quantitative extractions were achieved in SFE after addition of anhydrous sodium sulfate to the sample. Finally, ASE was applied to SPM-loaded filter candles whereby a mixture of n-hexane/acetone as extraction solvent allowed the simultaneous determination of PAHs, alkanes, and chlorinated hydrocarbons. PMID- 21639260 TI - Interface for coupling capillary electrophoresis to inductively coupled plasma and on-column concentration technique. AB - A new interface for coupling capillary electrophoresis (CE) to inductively coupled plasma (ICP) was developed. The interface was built outside and independent of the nebulizer and could be easily connected with a microconcentric nebulizer (MCN) as well as conventional pneumatic nebulizers. An on-column concentration technique was used to increase the sensitivity and to enhance the resolution of the system of capillary electrophoresis-inductively coupled plasma atomic emission spectrometry (CE-ICP-AES). By doing this, it was possible to analyze 1 MUg/mL of total chromium (prepared with K(2)Cr(2)O(7) and CrCl(3)) and 1 MUg/mL of copper consisting of Cu(2+) and Cu(EDTA)(2-) with good spectroscopic intensity and efficient separation. Detection limits of 18 elements for MCN-ICP AES coupled with CE were assessed by continuous sample introduction without applying high voltage and were found to be 1-4 times higher than those typically obtained by using MCN-ICP-AES for elemental analysis (without connection to the CE interface). When the on-column concentration technique was used, the sensitivities and separations were further improved by increasing the amount of sample. A simple electrolyte (0.05 M HNO(3)) and a large inner diameter capillary (150 MUm) could also be used to attain efficient separation. PMID- 21639261 TI - Molecular Baskets in Supercritical CO(2). AB - Calixarenes are synthetic macrocyclic compounds, described as "molecular baskets" as they possess high ionophoric selectivity and form inclusion complexes with many important ionic guests. In our initial work, hexameric and tetrameric tert butylcalixarenes, unfunctionalized at the lower rim, are shown to be separable on a diol column using supercritical fluid chromatography with methanol/chloroform modified CO(2) as mobile phase. The variation in capacity factors for these calixarenes was studied as a function of modifier composition. However, the solubility of these molecular baskets in unmodified supercritical CO(2) is enhanced by fluorination at the upper rim. For example, when p-allylcalix[4]arene is derivatized by a thiol-ene addition reaction with heptadecafluorodecanethiol, CF(3)(CF(2))(7)(CH(2))(2)SH, a solubility of >0.12 mol L(-)(1) in supercritical CO(2) is measured for the p-heptadecafluorodecylthio-n-propylcalix[4]arene at 60 degrees C and 200 atm. However, subsequent lower rim functionalization to form the tetrahydroxamate derivative, while reducing the solubility, allows supercritical fluid extraction of Fe(III) by the fluorinated calix[4]arene ligands to be studied as a function of temperature and pressure and monitored using UV/visible and atomic absorption spectrometry. In particular, the visible absorption spectra obtained for the extracted Fe(III)-calix[4]arene tetrahydroxamate complex, collected in dimethyl sulfoxide, are indicative of octahedral Fe(III) complexation in a manner similar to that displayed by water soluble siderophores. Studies on the efficiency and selectivity of Fe(III) extraction are also reported. PMID- 21639262 TI - Analysis of USP Organic Volatile Impurities and Thirteen Other Common Residual Solvents by Static Headspace Analysis. AB - Static headspace gas chromatography was investigated for the analysis of residual solvents in size-limited samples. The advantages of improved limits of detection at low ppm levels and decreased sample size requirements were realized. This methodology allows the measurement of 18 common residual solvents in 6 min using 1 mg or less of sample. Greater recovery of analytes from pure dissolution solvents without the use of salts became possible when smaller preparations of samples were combined with the use of smaller vials. Rapid equilibration of the static headspace sample preparations was an additional benefit of this modification. Optimized chromatography was developed to take advantage of the shorter equilibration time and to increase instrument productivity. PMID- 21639263 TI - Generating Sensor Diversity through Combinatorial Polymer Synthesis. AB - A new approach for rapid, simple generation of uniquely responding sensors for use in polymer-based sensor arrays has been developed. Polymerization reactions between different combinations of two starting materials have been found to lead to many new, unique sensors with responses not simply related to the proportion of the starting materials. This approach is demonstrated in two ways: (a) the use of discrete polymer sensing cones each comprised of a specific monomer combination and (b) the fabrication of a gradient sensor, containing all combinations between the starting and ending monomer concentrations. Gradient sensors were fabricated using two different binary monomer systems, with both systems showing regions of broadly diverse fluorescence responses to organic vapor pulses. PMID- 21639264 TI - Analysis of organic compounds by particle beam/hollow cathode atomic emission spectroscopy: determinations of carbon and hydrogen in amino acids. AB - A detailed evaluation of the analytical characteristics of a particle beam/hollow cathode glow discharge atomic emission spectroscopy (PB/HC-AES) system is described for applications in the area of organic sample analysis. The optimization of sample introduction, nebulization parameters, and glow discharge conditions was performed for the elemental analysis (focusing on C and H) of a group of amino acids. By use of a high-efficiency thermoconcentric nebulizer, analyte particles are introduced into a heated hollow cathode glow discharge source, in either flow injection or continuous-flow mode, for subsequent vaporization/atomization and excitation. Nebulization temperature, solvent composition, and liquid flow rate were studied to elucidate their roles in the ultimate analyte emission characteristics for organic compound analysis. The hollow cathode operating discharge current and gas pressure were optimized, with the general responses found to be similar to those for the case of metal analysis. Background interferences from solvent and additive media on carbon and hydrogen determinations were studied and substantially reduced. The analytical response curves for carbon and hydrogen present in amino acids were obtained using 200 MUL injection volumes, showing less than 10% RSD for replicate injections over a concentration range of 10-250 ppm, with detection limits of 3 and 1 ppm, respectively, for C (I) and H (I) emission. Subsequent studies of the response of carbon and hydrogen emission signal intensities to differences in amino acid stoichiometries suggest a capability of the PB/HC-AES system for the determination of empirical formulas based on H (I)/C (I) intensity ratios. PMID- 21639265 TI - Suppression of electroosmotic flow and prevention of wall adsorption in capillary zone electrophoresis using zwitterionic surfactants. AB - Addition of zwitterionic surfactants such as dodecyldimethyl(3 sulfopropyl)ammonium hydroxide, hexadecyldimethyl(3-sulfopropyl)ammonium hydroxide, and coco (amidopropyl)hydroxyldimethylsulfobetaine (Rewoteric AM CAS U) to an electrophoretic buffer suppress the electroosmotic flow by 50-90%. Onset of suppression occurs at approximately the critical micelle concentration of the surfactant. CAS U effectively suppresses the electroosmotic flow over the pH range 3-12. Addition of 2 mM CAS U to the electrophoretic buffer prevents adsorption of cationic proteins lysozyme, alpha-chymotrypsinogen A, cytochrome c, and ribonuclease A. Migration time reproducibility for these proteins is ~1% RSD within 1 day and 2-5% from day to day. Efficiencies in excess of 750 000 plates/m and recoveries of >80% were observed for protein injections from distilled water. Alternatively if 2 mM CAS U is added to samples, recoveries were quantitative, although efficiencies decreased to 325 000-600 000 plates/m. The natural electroosmotic flow of the capillaries is regenerated simply by rinsing with sodium hydroxide. PMID- 21639266 TI - Compositional effects in the retention of colloids by thermal field-flow fractionation. AB - The retention of polystyrene and silica colloids that have been chemically modified is measured in several aqueous carrier liquids. Retention levels are governed by particle size and composition but are also sensitive to subtle changes in the carrier. Size-based selectivities are higher in aqueous carriers compared to acetonitrile. In aqueous carriers, retention varies dramatically with the nature of the additive, and for a given additive, retention increases with ionic strength, regardless of modifications to the particle surface. The role played by electrostatic effects in retention is studied by varying the ionic strength of the carrier, estimating electrical double layers, determining particle-wall interaction parameters, and calculating the coefficients of mass diffusion and thermal diffusion. Although electrostatic phenomena can affect mass diffusion and particle-wall interactions in carriers of low ionic strength (<10( 3) M), such effects are not great enough to explain the dependence of retention on ionic strength. Therefore, thermal diffusion must be affected directly. Thermal diffusion is found to increase with pH, and at a given pH with the surface tension of the suspended particle. Finally, while the addition of the surfactant FL-70 generally decreases retention, greater retention levels can ultimately be achieved with FL-70 because larger temperature gradients can be used without particle adsorption to the accumulation wall. PMID- 21639267 TI - Integrated capillary electrophoresis on flexible silicone microdevices: analysis of DNA restriction fragments and detection of single DNA molecules on microchips. AB - Microchips for integrated capillary electrophoresis systems were produced by molding a poly(dimethylsiloxane) (PDMS) silicone elastomer against a microfabricated master. The good adhesion of the PDMS devices on clean planar surfaces allows for a simple and inexpensive generation of networks of sealed microchannels, thus removing the constraints of elaborate bonding procedures. The performance of the devices is demonstrated with both fast separations of phiX 174/HaeIII DNA restriction fragments labeled with the intercalating dye YOYO-1 and fluorescently labeled peptides. Detection limits in the order of a few zeptomoles (10(-)(21) mol) have been achieved for each injected DNA fragment, corresponding to a mass detection limit of ~2 fg for the 603 base pair fragment. Single lambda-DNA molecules intercalated with YOYO-1 at a base pair-to-dye ratio of 10:1 could be detected with an uncomplicated laser-induced fluorescence detection setup. High single-molecule detection efficiency (>50%) was achieved under electrophoretically controlled mass transport conditions in PDMS microchannels. PMID- 21639268 TI - On-the-Fly Fluorescence Lifetime Determination with Total Emission Detection in HPLC. AB - The entire fluorescence decay profile during HPLC elution has been directly measured on-the-fly in HPLC at higher sensitivity than in previous literature reports. The fluorescence is excited with the fourth harmonic (266 nm) of a pulsed Nd:YAG laser system and detected broadband with a photomultiplier tube and a digital storage oscilloscope. Detection limits in the range 1-10 ppb are found for several individual polycyclic aromatic hydrocarbons (PAHs) when the total time-integrated fluorescence is analyzed. The chromatograms of PAH mixtures containing 8-10 species were lifetime analyzed with a simple phase plane analysis, in which a single lifetime is determined from the fluorescence decay profile for each point on the chromatogram. The determination of lifetimes under coelution conditions is also illustrated and discussed. PMID- 21639269 TI - Simultaneous flame ionization and absorbance detection of volatile and nonvolatile compounds by reversed-phase liquid chromatography with a water mobile phase. AB - A flame ionization detector (FID) is used to detect volatile organic compounds that have been separated by water-only reversed-phase liquid chromatography (WRP LC). The mobile phase is 100% water at room temperature, without use of organic solvent modifiers. An interface between the LC and detector is presented, whereby a helium stream samples the vapor of volatile components from individual drops of the LC eluent, and the vapor-enriched gas stream is sent to the FID. The design of the drop headspace cell is simple because the water-only nature of the LC separation obviates the need to do any organic solvent removal prior to gas phase detection. Despite the absence of organic modifier, hydrophobic compounds can be separated in a reasonable time due to the low phase volume ratio of the WRP-LC columns. The drop headspace interface easily handles LC flows of 1 mL/min, and, in fact, compound detection limits are improved at faster liquid flow rates. The transfer efficiency of the headspace interface was estimated at 10% for toluene in water at 1 mL/min but varies depending on the volatility of each analyte. The detection system is linear over more than 5 orders of 1-butanol concentration in water and is able to detect sub-ppb amounts of o-xylene and other aromatic compounds in water. In order to analyze volatile and nonvolatile analytes simultaneously, the FID is coupled in series to a WRP-LC system with UV absorbance detection. WRP-LC improves UV absorbance detection limits because the absence of organic modifier allows the detector to be operated in the short wavelength UV region, where analytes generally have significantly larger molar absorptivities. The selectivity the headspace interface provides for flame ionization detection of volatiles is demonstrated with a separation of 1-butanol, 1,1,2-trichloroethane (TCE), and chlorobenzene in a mixture of benzoic acid in water. Despite coelution of butanol and TCE with the benzoate anion, the nonvolatile benzoate anion does not appear in the FID signal, allowing the analytes of interest to be readily detected. The complementary selectivity of UV visible absorbance detection and this implementation of flame ionization detection allows for the analysis of volatile and nonvolatile components of complex samples using WRP-LC without the requirement that all the components of interest be fully resolved, thus simplifying the sample preparation and chromatographic requirements. This instrument should be applicable to routine automated water monitoring, in which repetitive injection of water samples onto a gas chromatograph is not recommended. PMID- 21639270 TI - LC-Biosensor System for the Determination of the Neurotoxin beta-N-Oxalyl-l alpha,beta-diaminopropionic Acid. AB - An analysis system is described for the determination of the neurotoxin beta-N oxalyl-l-alpha,beta-diaminopropionic acid (beta-ODAP). The system is based on liquid chromatographic separation of beta-ODAP from interfering amino acids on an anion exchange column coupled with an amperometric enzyme electrode for the registration of beta-ODAP. The electrode is based on a graphite rod modified with an Os(2+/3+) redox polymer cross-linked with l-glutamate oxidase and horseradish peroxidase. This LC-bienzyme electrode analytical system enabled monitoring of as low as 4 MUM beta-ODAP (injection volume 100 MUL). The beta-ODAP content in real grass pea samples was measured to range between 3.3 and 5.2 g kg(-)(1) in dry grass pea. PMID- 21639271 TI - Critical examination of the quantification of aromatic compounds in three standard reference materials. AB - Three standard reference materials from the National Institute of Standards and Technology (NIST), namely, SRM 1597 coal tar, 1582 crude oil, and 1580 shale oil, were investigated in detail to determine the concentration of polycyclic aromatic sulfur heterocycles using gas chromatography with the atomic emission detector in the carbon- and sulfur-selective modes. Coelution problems were found to be common, and the use of two capillary gas chromatographic columns with stationary phases of widely differing polarity was necessary for the separation of the important analytes phenanthrene, anthracene, dibenzothiophene, and the naphthothiophenes. On the commonly used nonpolar stationary phases, phenanthrene and dibenzothiophene coelute with the isomeric naphthothiophenes and this leads to too high concentrations being measured for the two major analytes and must be corrected for. For (sulfur and oxygen) heterocycles, individual response factors must be used if the flame ionization detector is employed. The NIST values were obtained without regard to those factors. This is done here for the three SRMs, and it is shown that the adjusted NIST values agree very well with the GC/AED values. It is suggested that the (noncertified) NIST values for several polycyclic aromatic compounds (PACs) should be reexamined. Very probably many other determinations of PACs might suffer from the same shortcomings. PMID- 21639272 TI - Solvatochromic Study of Poly(vinyl chloride) Plasticizers and Their Solutions in Chloroform: Application to Phenobarbital Partitioning and Molecular Recognition of Phenobarbital. AB - The possibility now exists, with the availability of several families of artificial molecular receptors, to create selective extraction media. More selective extractions will lead to cleaner chromatograms, with lower detection limits and perhaps higher accuracy for trace organic analysis by chromatography. Furthermore, laboratories will be expected to minimize the use of volatile organic solvents. Consequently, nonvolatile, reusable solvents will be the basis for extractions. In addition, as artificial molecular receptors become more widely available, these solvents will be used to support molecular recognition. We have focused on plasticizers of poly(vinyl chloride) as examples of these solvents. We have determined solvatochromic parameters of several plasticizers and their solutions in chloroform. These parameters, along with cohesive energy density and solvent molar volume, were used to derive linear free energy relationships for the free energies of phenobarbital partitioning between solvent and aqueous solution, receptor solubility, formation of a complex with a barbiturate receptor [1,3-bis[[[6-(1-butyrylamino)pyrid-2 yl]amino]carbonyl]benzene (2)], and the transfer of the complex (artificial receptor and phenobarbital) from chloroform to other solvents. Solvent dipolarity/polarizability and hydrogen bond basicity, but not acidity, support complex dissociation. Solvents with large molar volumes dissolve the polar solutes, phenobarbital, receptor, and complex more poorly than solvents with lower molar volume, but there is no influence of molar volume on complex formation. PMID- 21639273 TI - Dynamic surface tension and adhesion detection for the rapid analysis of surfactants in flowing aqueous liquids. AB - A multidimensional dynamic surface tension detector (DSTD) for flowing liquid samples is reported. The DSTD is based on measuring the pressure with time of a repeating drop formed by the flow of liquid out the end of a pointed stainless steel capillary. This pressure-based DSTD provides information on dynamic surface tension at the air-liquid interface and adhesion at the solid-liquid interface on the side of the pointed capillary tip for each drop of surfactant solution, resulting in rapid characterization of complex samples. The signal obtained with the pressure-based DSTD is characterized, and a method is developed for extraction of the desired analytical information from the pressure signal. The DSTD was calibrated with 2-propanol over the range of 0-0.30 in relative surface tension lowering, Deltagamma/gamma. The experimentally obtained Deltagamma/gamma was in agreement with a theoretical model and published data for Deltagamma/gamma over this range. A data analysis method was developed to adapt the DSTD to applications such as liquid chromatography and flow injection analysis, where the concentration of surfactant changes as a function of time. The DSTD signal yields a pressure-based Deltagamma/gamma that is due to surface tension alone and a time based Deltagamma/gamma that is a combination of both surface tension and adhesion, providing essentially a contact angle measurement on a flowing sample. The data analysis method involves plotting the pressure-based and time-based surface tension measurements against each other at the same surfactant concentration for each pair of measurements, yet over a range of concentrations to establish a slope. This is referred to as a dynamic analysis plot and is applied in the characterization of various surfactants such as dodecyl sulfate ion-paired with tetrabutylammonium, industrial surfactant solutions FC-171 and FC 129, and biological surfactants tetradecyl maltoside, benzyldimethyldodecylammonium bromide, and 3-(N-decyl-N,N-dimethylammonio)-1 propanesulfonate. The slopes of the dynamic analysis plots for these surfactants were found to be unique, generally independent of concentration, and useful for understanding the type and degree of operating surface interactions. The FC-171 solution was found to exhibit considerable adhesion at the capillary tip, while dodecyl sulfate was found to have a small adhesion effect. Adhesion for dodecyl sulfate solutions was significantly enhanced by coating the stainless steel capillary tip with a hydrophobic polymer. Thus, there is potential for tuning the extent of the surface tension and adhesion effects for selective chemical analysis. The detection limit for dodecyl sulfate ion-paired with tetrabutylammonium is 0.9 ppm. Application of the DSTD for liquid chromatography is demonstrated, and the multidimensional data are shown to be useful in identifying and characterizing the poly(ethylene glycol)s separated from 2 propanol. PMID- 21639274 TI - Pulsed-accelerated-flow spectrometer with position-resolved observation. AB - The title instrument (PAF-PRO) permits the progress of rapid reactions to be monitored at 128 positions along a 2 cm observation cell as reactants flow down the cell. A decelerated push is used to give velocity changes from 21 to 2 m s( 1) during a time interval of 384 ms. Variation of flow velocity allows reaction rate constants to be resolved from the physical mixing process. The flow system brings the reacting solutions together in a 10-jet radial mixer 0.32 cm before the mixture enters the observation tube (0.203 cm width, 1.945 cm length). A masked charge-coupled device (CCD) is the array detector used to obtain transmittance data as a function of position and velocity. The masked portion of the CCD serves as a dynamic memory buffer for fast data acquisition. Pseudo-first order rate constants are measured from 200 to 12 000 s(-1). The instrument also is calibrated for second-order reactions (equal concentrations) with initial half lives of 0.3-1.5 ms. Applications of the PAF-PRO system for the study of fast multistep reactions are presented. PMID- 21639275 TI - Graphite-Poly(tetrafluoroethylene) Composite Enzyme Electrodes as Suitable Biosensors in Predominantly Nonaqueous Media. AB - The performance of a graphite-poly(tetrafluoroethylene) Teflon composite amperometric ferrocyanide-mediated peroxidase electrode in a predominantly nonaqueous medium such as reversed micelles is discussed and compared with the behavior in a medium formed by acetonitrile/water. The composite electrode was constructed by purely physical entrapment of both the enzyme and the mediator into the bulk of the graphite-Teflon matrix with no need of covalent attachments. This biosensor responded rapidly to the changes in the concentration of both hydrogen peroxide and 2-butanone peroxide in reversed micelles formed with ethyl acetate, 0.1 mol L(-)(1) dioctyl sulfosuccinate as the surfactant, and a 4% phosphate buffer (pH 7.4) as the dispersed phase. The electrode showed a long term operation due to the renewability of its surface. Moreover, reproducible responses were obtained with different electrodes fabricated from different composite matrixes. No significant loss of the enzyme activity was observed after four months of dry storage at 4 degrees C of the composite electrode. Limits of detection of 2.1 * 10(-)(7) and 3.5 * 10(-)(7) mol L(-)(1) were obtained for H(2)O(2) and 2-butanone peroxide, respectively. The possibility of using this biocomposite electrode in flowing systems, using the reversed micelles as the carrier, has been demonstrated. The kinetic of the enzymatic reaction was faster in a 90:10 acetonitrile/phosphate buffer medium than in reversed micelles, which can be attributed to the higher water content present in the former medium. A similar stability of the biosensor and a slightly better sensitivity for peroxides was observed in the acetonitrile/water mixture when compared with reversed micelles. Finally, the electrode also performed well in the flow injection mode. PMID- 21639276 TI - A Microscale NO(3)(-) Biosensor for Environmental Applications. AB - A biosensor for NO(3)(-) containing immobilized dentrifying bacteria and a reservoir of liquid growth medium for the bacteria was constructed. The bacteria did not have a N(2)O reductase and therefore reduced NO(3)(-) to N(2)O, which was then subsequently quantified by a built-in electrochemical transducer for N(2)O. The only agents interfering with the determination of NO(3)(-) were NO(2)(-) and N(2)O. The sensitivity for NO(2)(-) was identical to the one for NO(3)(-) whereas the sensitivity for N(2)O was 2.4 times higher than for NO(3)(-). Diffusive supply of electron donors to the bacteria from the built-in reservoir of growth medium ensured that the biosensor could work for 2-4 days. The tip diameter was down to 20 MUm, and the sensors exhibited perfectly linear responses to nitrate in both freshwater and seawater. The detection limit was ~1 MUM. The 90% response time to changes in NO(3)(-) concentration was from 15 to 60 s at room temperature and about twice that at 6 degrees C, which was the lowest temperature for successful operation. The new NO(3)(-) biosensor is a very useful tool for the study of nitrogen metabolism in nature. PMID- 21639277 TI - Porphyrin-modified electrodes as biomimetic sensors for the determination of organohalide pollutants in aqueous samples. AB - A preliminary examination of a simple and rapid screening method for quantifying a range of toxic organohalides directly in aqueous solution based on their electrocatalytic reduction with a metalloporphyrin catalyst is described. Homogenous catalysis is described as well as heterogeneous catalysis using precipitated cobalt(II) tetraphenylporphine ((TPP)Co) at a graphite foil electrode which permitted the sensitive detection of a wide range of different organohalides, including a number of chemically diverse industrial pollutants such as 1,2,3,4,5,6-hexachlorocyclohexane (lindane) and carbon tetrachloride, representative of haloalkane compounds, haloalkenes such as perchloroethylene, and aromatics, such as 2,4-dichlorophenoxyacetic acid, pentachlorophenol, and the insecticide DDT. The coordinating effect of solvent on the thermodynamics of the Co(II)/(I) electrode reaction is used to practical advantage to build an amperometric detector that is insensitive to interference from oxygen, a parameter that varies considerably in environmental samples. Devices also appear relatively insensitive to the ionic composition of the analyte sample. The work provides the basis for developing a prototype sensor for screening toxic organohalogen pollutants for use in environmental monitoring situations. PMID- 21639278 TI - Anodic stripping voltammetry coupled on-line with inductively coupled plasma mass spectrometry: optimization of a thin-layer flow cell system for analyte signal enhancement. AB - Parameters affecting analyte signal enhancement in anodic stripping voltammetry inductively coupled plasma mass spectrometry (ASV-ICP-MS), using a thin-layer ASV cell and microconcentric nebulization (MCN), have been examined. Silver was used as a test analyte and was deposited at a glassy carbon working electrode. The MCN allowed use of solution flow rates that were beneficial to optimum electrolytic performance of the thin-layer cell. High analyte deposition efficiencies obtained with the thin-layer cell, combined with minimal sample consumption of the MCN, allowed substantial signal enhancement (>400 times higher than continuous nebulization level) to be obtained with 2-3 mL of sample and deposition times of less than 30 min. Signal enhancement was strongly influenced by the opposing effect of flow rate on the electrolytic deposition efficiency (deposition efficiency decreases with increasing flow rate) and on the quantity of analyte delivered to the cell (analyte mass throughput increases with increasing flow rate). Excellent linearity for stripping peak heights was demonstrated for a wide range of analyte deposition times and for peak heights and peak areas (r > 0.999) over a wide concentration range (25 ng/L-20 MUg/L). Precision was good (RSD typically <3% for n = 3-6) except for a high Ag blank contributed to by corrosion of the counter electrode and by Ag diffusion from the reference electrode into the cell. Details of the flow manifold and ASV cells are discussed, along with relevant performance characteristics of the MCN. PMID- 21639279 TI - Direct electrochemical detection of purine- and pyrimidine-based nucleotides with sinusoidal voltammetry. AB - Sinusoidal voltammetry was employed to detect both purine- and pyrimidine-based nucleic acids. Adenine and cytosine, representing these two classes of nucleic acids, could be measured with submicromolar detection limits at a copper electrode under these conditions, where the sensitivity for adenine was much higher than that for cytosine. Detection limits for purine-containing nucleotides [e.g., adenosine 5'-monophosphate (AMP), adenosine 5'-diphosphate (ADP), and adenosine 5'-triphosphate (ATP)] were on the order of 70-200 nM using this method. These detection limits are achieved for native nucleotides and are over 2 orders of magnitude lower than those found with UV absorbance detection. Submicromolar detection limits were also obtained for pyrimidine-based nucleotides, which could also be detected with high sensitivity due to the presence of a sugar backbone that is electroactive at the copper surface. This detector is not fouled by the nucleotides and may be used for the sensitive detection of analytes eluting continuously in a flowing stream, i.e., from a chromatography column or an electrophoresis capillary. PMID- 21639280 TI - Determination of extractable organic chlorine and bromine by probe injection dual microplasma atomic emission spectrometry. AB - A probe injection dual-microplasma spectrometer is evaluated as a low-cost alternative for the determination of extractable organic chlorine and bromine (EOCl and EOBr). The system consists of two 350 kHz plasmas sustained in the same stream of helium and a probe for sample application in the interplasma region. The sample was applied with a microsyringe into a small cup on the sample probe. Subsequently, the extraction solvent was gently evaporated, and the sample cup was pushed into the interplasma region. The first plasma was in direct contact with the sample probe and served to rapidly vaporize the sample material. The vaporized sample was then transferred to the second plasma, where atomic emission was measured for the determination of EOCl and EOBr. For both Cl and Br, 120 pg detection limits and 1000:1 halogen-to-carbon selectivities were obtained, and responses were linear over 3 orders of magnitude. PMID- 21639281 TI - Continuous-flow extraction of colloidal components in aqueous samples. AB - In this study we report the development and performance of a system for continuous-flow extraction of dissolved and colloidal analytes in an aqueous matrix. Initial studies, using a classical segmented-flow extraction procedure, showed poor extraction efficiency for the hydrophobic colloidally dispersed analytes. Insufficient contact between the extractant and the colloidal constituents seems to be the primary reason for poor extraction. Improved performance is obtained when mechanical energy is added to the system, to effect a forced contact between the sample and the solvent. This was accomplished by injecting the extractant, with a high velocity, into the continuous flow of analyte through a narrow-bore nozzle. In this way, the solvent stream is dispersed into fine droplets with high kinetic energies. A region of intense turbulence is created, which was studied by high-speed photography using pulsed laser fluorescence. Comparison with classical flow extraction, using a model sample of colloidal wood resin compounds in water, showed that the dissolved components extracted well with both systems, while an extraction enhancement of up to 9 times was experienced with colloidal triglycerides. PMID- 21639282 TI - A microwave-powered thermospray nebulizer for liquid sample introduction in inductively coupled plasma atomic emission spectrometry. AB - A new thermospray nebulizer based on the absorption of microwave radiation (MWTN) by aqueous solutions of strong acids is presented for the first time. To this end, a given length of the sample capillary is placed inside the cavity of a focused microwave system. A small piece of a narrower capillary tubing is connected at the tip of the sample capillary, outside the microwave cavity, to build up pressure. Drop size distributions of primary aerosols are exhaustively measured in order to evaluate the influence of several experimental variables (microwave power, liquid flow, irradiation length, inner diameter of the outlet capillary, nature and concentration of the acid) on the characteristics of the primary aerosol that are related to the emission signal. These experiments have been performed mainly to increase our understanding of the microscopic process of this new type of aerosol generation. A standard Meinhard nebulizer was employed for comparison. Under the best conditions the entire aerosol volume is contained in droplets smaller than 20 MUm compared with 45% of the volume of the aerosol generated by the Meinhard. Hence, higher analyte and aerosol transport rates are to be expected for the MWTN compared with the Meinhard nebulizer. As any highly efficient nebulizer, MWTN requires a desolvation unit. For solutions 0.75 M in strong acid, the new nebulizer improves sensitivity (1.0-2.8 times), limits of detection (1.2-3.0 times), and background equivalent concentration (0.9-2.0 times) as compared to the standard Meinhard nebulizer, features many of the advantages of the conventional thermospray nebulizer, and overcomes some of its drawbacks (MWTN does not show corrosion problems and works at lower pressure, the aerosol characteristics are not modified when the PTFE capillary is replaced). PMID- 21639283 TI - Cation selectivity of natural and synthetic ionophores probed with laser-induced liquid beam mass spectrometry. AB - The novel laser desorption method laser-induced liquid beam ionization/desorption (LILBID) is applied to the mass spectrometric examination of selective ion binding by natural and synthetic ionophores in methanol solutions. The ions are desorbed from a liquid jet with an IR laser pulse and then extracted perpendicularly into a reflectron time-of-flight (RE-TOF) analyzer. LILBID studies on the natural ion carriers valinomycin and monensin A are presented, as well as those on the synthetic crown ethers 18-crown-6, diaza-18-crown-6, and benzo-15-crown-5. No fragment ions are detected, and the measured ion selectivity is in good qualitative agreement with published stability constants of the complexes. The observed specific recognition of silver ions by diaza-18-crown-6 can be rationalized by the principle of hard and soft acids and bases, which predicts stable complexes when the polarizabilities of Lewis acid and base are similar. Weak, noncovalent interactions like those in the sandwich complex between two benzo-15-crown-5 molecules with one potassium ion are detected with LILBID. Their preservation during the process of ion desorption depends on the laser intensity. A comparison with spectra obtained by using electrospray ionization (ESI) and matrix assisted laser desorption/ionization (MALDI) shows that LILBID can potentially become a sensitive tool for the screening of weak but specific molecular interactions. PMID- 21639284 TI - Ultrahigh Mass Accuracy in Isotope-Selective Collision-Induced Dissociation Using Correlated Sweep Excitation and Sustained Off-Resonance Irradiation: A Fourier Transform Ion Cyclotron Resonance Mass Spectrometry Case Study on the [M + 2H](2+) Ion of Bradykinin. AB - Using electrospray ionization with a 9.4 T Fourier transform mass spectrometer, fragment ion spectra were acquired for a single isotopomer of doubly protonated bradykinin (molecular mass, 1059.6 Da). Correlated sweep excitation methods were applied to mass-select the single isotopomer (m/z = 530.8). Sustained off resonance irradiation was used to activate and fragment the ions. The accuracy (in terms of m/z) in detection of the fragment ions was on average 1.2 ppm, making the assignments unambiguous. The methods employed would be generally applicable to ions in the mass range of approximately 50 Da to 50 kDa. PMID- 21639285 TI - A Laminar Flow Nebulizer for Aerosol MALDI. AB - A laminar flow nebulizer was developed for use with aerosol matrix-assisted laser desorption/ionization mass spectrometry. The nebulizer consists of a glass pneumatic nebulizer combined with a laminar flow tube. Particles formed at the pneumatic nebulizer are entrained in an auxiliary gas stream that dries the particles in the laminar flow tube. The mass range of the reflectron time-of flight mass spectrometer has been extended to greater than 5000 Da: a mass spectrum of bovine insulin is reported at a mass resolution of 100. Mass spectra of bovine insulin and the peptides bradykinin and angiotensin II contain features resulting from prompt analyte dissociation prior to ion acceleration. PMID- 21639286 TI - 1-Methyl-1-(2,4-dinitrophenyl)hydrazine as a New Reagent for the HPLC Determination of Aldehydes. AB - The synthesis and first application of an N-alkylated hydrazine reagent for the HPLC determination of aldehydes and ketones is described. 1-Methyl-1-(2,4 dinitrophenyl)hydrazine (MDNPH) reacts with aldehydes to give the corresponding hydrazones in the presence of an acid as catalyst. In contrast to other hydrazine reagents, MDNPH is oxidized by both ozone and nitrogen dioxide quantitatively to N-methyl-2,4-dinitroaniline (MDNA), which can be separated from the hydrazones of the lower aldehydes by means of HPLC. Unexpected elution orders are observed for the 1-methyl-1-(2,4-dinitrophenyl)hydrazones compared to those of 2,4 dinitrophenylhydrazones. Dual-wavelength detection is employed as a means for identification of different groups of the hydrazones and MDNA. PMID- 21639287 TI - Simultaneous HPLC determination of peroxyacetic Acid and hydrogen peroxide. AB - The first instrumental method for simultaneous determination of peroxyacetic acid (PAA) and hydrogen peroxide has been developed. The successive quantitative reaction of PAA with methyl p-tolyl sulfide (MTS) and hydrogen peroxide with triphenylphosphine (TPP) yields the corresponding sulfoxide MTSO and phosphine oxide TPPO. The reagents and their oxides are separated by HPLC on reversed-phase columns with acetonitrile/water gradient elution within 5 min. External calibration with the solid standards of MTSO and TPPO leads to a very accurate and reliable method. Samples are stable and can be stored after derivatization for several days prior to analysis. Real samples from brewery disinfection were analyzed in comparison to titration with excellent correlation. PMID- 21639288 TI - Determination of iodide in seawater by ion chromatography. AB - A simple and highly sensitive ion chromatographic method with UV detection was developed for iodide (I(-)) in seawater. A high-capacity anion-exchange resin with polystyrene-divinylbenzene matrix was used for both preconcentration and separation of iodide. Iodide in artificial seawater (salinity, 350/00) was trapped quantitatively (98.8 +/- 0.6%) without peak broadening on a preconcentrator column and was separated with 0.35 M NaClO(4) + 0.01 M phosphate buffer (pH 6.1). On the other hand, the major anions in seawater, chloride and sulfate ions, were partially trapped (5-20%) and did not interfere in the determination of I(-). The detection limit for I(-) was 0.2 MUg/L for 6 mL of artificial seawater. The present method was applied to determination of I(-) (ND 18.3 MUg/L) and total inorganic iodine (I(-) + IO(3)(-) - I, 50.0-52.7 MUg/L) in seawater samples taken near Japan. PMID- 21639289 TI - Development of a detector for ultratrace nitrogen in argon using low-pressure, capillary glow discharge molecular emission spectrophotometry. AB - A highly sensitive detector was developed for determining parts per billion of nitrogen in high-purity argon. The method is based on the spectrophotometric determination of the emission intensity from the nitrogen molecule excited in the lowest vibrational transitions of the electronic C(3)Pi(u) -> B(3)Pi(g) system. A capillary glow discharge technique was applied to excite nitrogen in an argon flow at several Torr. The low-pressure, capillary glow discharge method offered high sensitivity for detecting trace nitrogen in argon. The response of the detector was proportional in the range of nitrogen concentrations from 0.1 ppb, which correspond to S/N = 3, to 100 ppm. PMID- 21639290 TI - Estimation of Free Radical Ionization Energies by the Kinetic Method and the Relationship between the Kinetic Method and the Hammett Equation. AB - Substituted 1,2-diphenylethanes undergo competitive dissociations upon electron ionization (EI) to generate substituted benzyl cation and benzyl radical pairs. Application of the kinetic method to the previous reported EI mass spectra of these covalently bound precursor ions (data are taken from McLafferty et al. J. Am. Chem. Soc. 1970, 92, 6867)) is used to estimate the ionization energies of substituted benzyl free radicals. A correlation is observed between the Hammett sigma constant of the substituents and the kinetic method parameter, ln(k(x)/k(H)), where k(x) is the rate of fragmentation to give the substituted product ion and k(H) is the rate to give the benzyl ion itself. Systems involving weakly bound cluster ions, including proton-bound dimers of meta- and para substituted pyridines and meta- and para-substituted anilines, and electron-bound dimers of meta- and para-substituted nitrobenzenes, also show good correlations between the kinetic method parameter and the Hammett sigma constant. PMID- 21639291 TI - Immunoaffinity Ultrafiltration with Ion Spray HPLC/MS for Screening Small Molecule Libraries. AB - A solution-phase screening method for libraries of pharmaceutically relevant molecules is presented. The technique is applicable to screening combinatorial libraries of 20-30 closely related molecules. In this report, individual benzodiazepines are selected from a multicomponent library mixture by formation in solution of noncovalent immunoaffinity complexes with antibodies raised to therapeutically proven drugs such as nitrazepam, temazepam, or oxazepam. Captured compounds are separated from nonspecifically bound library components by centrifugal ultrafiltration. The specifically selected molecules retained on the filter are subsequently liberated from the antibodies by acidification and analyzed by HPLC coupled with pneumatically assisted electrospray (ion spray) ionization mass spectrometric detection. Competition by the benzodiazepines for limited antibody binding sites is controlled by varying the stoichiometry of the complexation mixture. This procedure selects library components with the greatest affinity for a particular antibody. Specific capture of benzodiazepines is demonstrated by screening both a pool of structurally similar benzodiazepines and a more complex mixture of benzodiazepines with an additional set of unrelated compounds. Affinity ultrafiltration and electrospray mass spectrometry complement each other to enhance screening and identification of pooled drug candidates and potentially can be extended to other small-molecule combinatorial libraries and macromolecular receptor preparations. PMID- 21639292 TI - Analysis of naturally produced technetium and plutonium in geologic materials. AB - In uncontaminated natural materials, plutonium and technetium exist exclusively as products (daughters) of nuclear reactions in which uranium is the principal reactant (parent). Under conditions of chemical stability over geologic periods of time, the relative abundances of daughter and parent elements are fixed by the rates of nuclear reactions and the decay of the daughter radionuclide. The state of this nuclear secular equilibrium condition is the primary basis of the geochemical study of these elements in nature. Thus, it is critical that nuclear parent and daughter abundances are measured in the same sample. We have developed a quantitative procedure for measuring subpicogram quantities of plutonium and technetium in gram quantities of geologic matrices such as uranium ores. The procedure takes advantage of the aggressive properties of sodium peroxide/hydroxide fusion to ensure complete dissolution and homogenization of complex materials, the precision provided by isotope dilution techniques, and the extreme sensitivity offered by thermal ionization mass spectrometry. Using this technique, a quantitative aliquot can be removed for uranium analysis by isotope dilution thermal ionization mass spectrometry or alpha spectrometry. Although the application of the procedure is unique, the analytical concepts may find more general application in studies of environmental contamination by nuclear materials. To assess the precision and accuracy of the analytical results, blanks and standards were analyzed routinely for a 1-year period to ensure quality control of our sample analyses. The average technetium blank is 5 +/- 4 fg (n = 8), and that for plutonium is 0.17 +/- 0.15 pg (n = 7). Thus, the detection limit for technetium (defined as 3 times the standard deviation of the average blank) is 11 fg, and that for plutonium is 0.44 pg. To assess the procedural precision, Canadian Reference Material BL-5 was analyzed routinely with samples. The results of seven replicate analyses for technetium in this standard reference material yield a technetium concentration of 59.0 fg/g, with a remarkably small standard deviation of 0.6 fg, 1.0% of the average value. The results of six replicate analyses for the concentration of plutonium in BL-5 give 1.012 pg/g, with an equally small standard deviation of 0.016, 1.6% of the average value. No direct measure of accuracy can be done on the technetium or plutonium analyses, because no standard reference material exists for these elements. To help constrain the accuracy of our measurements, equilibrium technetium/uranium and plutonium/uranium abundances were calculated using the nuclear reaction code MCNP. For technetium, such calculations are relatively insensitive to variations in model parameters, and measurements fall within a 21% high/low bias. For plutonium, the calculations are very sensitive to model parameters and hence inherently less precise. Indirectly, spike and isotope mix calibrations made from weighted quantities of certified isotopes (both technetium and plutonium) can be used to determine the bias of the measurement system for these elements. These calibrations show that the measurement system is biased by no more than +/-1.5%. PMID- 21639293 TI - Distinction of amino Acid enantiomers based on the basicity of their dimers. AB - Mixtures of several amino acid pairs, in all four chiral combinations, were studied. The protonated trimers (A(2)BH(+)) fragment, forming ABH(+) and A(2)H(+) dimers. Abundance ratios of these fragments were measured in the mass-analyzed ion kinetic energy spectra of the trimers. These were found to depend on the stereochemistry (homo- or heterochiral form) of the ABH(+) dimer. The results were evaluated using the kinetic method, and the chiral discrimination was related to a difference in gas-phase basicity (GB) between the homo- and the heterochiral dimers. Four amino acid pairs (proline-tryptophan, phenylalanine alanine, phenylalanine-proline, and phenylalanine-valine) were studied. Chiral discriminations were observed in all cases, relating to 0.4-4 kJ/mol differences in GB. The technique described here can generally be used to study enantiomers by mass spectrometry and is capable of reliably distinguishing energy differences as small as 0.2 kJ/mol in cluster ions. PMID- 21639294 TI - Time-of-Flight Mass Spectrometry with a Pulsed Glow Discharge Ionization Source. AB - The pulsed glow discharge (GD) plasma source exhibits several characteristics that make it ideally suited for use with time-of-flight mass spectrometry (TOFMS). TOFMS uniquely affords the ability to monitor a narrow temporal window for a time-varying process such as ion formation in the pulsed glow discharge plasma. Pulsed GD-TOFMS exhibited distinct advantages for the direct determination of trace elements in solid state samples. Initially, the pulse powered GD-TOFMS system used for these investigations exhibited poor resolution. In an effort to improve resolution, a slit was introduced to narrow the ion beam orthogonally entering the extraction region of the TOFMS. In an effort to determine optimal operating conditions, the influence of slit width on TOF performance was investigated. In the course of this study, the slit width was found to influence isotope ratio accuracy as well as resolution. A slit width of 1.0 mm was determined to provide the best compromise between resolution and isotope ratio accuracy. Pulsed GD-TOFMS affords improved sensitivity and selectivity because Penning ionization is enhanced during the time period immediately following the termination of the discharge power. Ions sampled by an extraction pulse applied after power termination also yield a mass spectrum that is free of contributions arising from electron-ionized interferences. This advantage arises because only ions generated via the Penning ionization mechanism persist after the termination of discharge power. Sampling in the "afterpeak" time regime eliminates the saturation of the detector arising from discharge support gas ion signal. PMID- 21639295 TI - Quantitative determination of the components in overlapping chromatographic peaks using wavelet transform. AB - A novel application of the wavelet transform in retrieving the separate signals from overlapping chromatographic peaks and quantitative determination of the components in the overlapping chromatograms is described. The signals can be very easily separated by decomposing an overlapping chromatographic peak into localized contributions according to their frequency, and quantitative calculation can be done by studying the contributions of higher frequency. Overlapping peaks of two- and three-component mixtures were investigated by the method, and the results show excellent correlations between peak areas of the retrieved signals and the concentrations for all of the components. PMID- 21639296 TI - Separation and determination of polycyclic aromatic hydrocarbons by solid phase microextraction/cyclodextrin-modified capillary electrophoresis. AB - A sensitive method for the determination of polycyclic aromatic hydrocarbons (PAHs) by solid phase microextraction coupled with cyclodextrin (CD)-modified capillary electrophoresis (CE) using UV detection has been developed. A glass fiber was prepared and used for absorbing 16 EPA priority PAHs from diluted samples until equilibrium was reached. After the glass fiber was connected to a separation capillary via an adapter, the absorbed analytes were directly released into the CE buffer stream, and electrophoretic separation was effected using a 50 mM borate, pH 9.2, buffer containing 35 mM sulfobutyloxy-beta-CD, 10 mM methyl beta-CD, and 4 mM alpha-CD. Separation was effected since neutral PAHs differentially partitioned between the neutral and charged CD phases. Under 30 kV applied potential, separation was achieved in less than 15 min with high resolution and number of theoretical plates. Pyrene as low as 8 ppb was detected, while the highest limit of detection was 75 ppb for acenaphthene. Very satisfactory reproducibility with respect to migration time and peak area was obtained for repetitions using the same separation capillary and adapter, where only the extraction fiber was discarded after each analysis. PMID- 21639297 TI - A glow discharge-conductivity detector for carbohydrates in aqueous chromatography. AB - A new glow discharge-conductivity detector is shown to respond to low nanomolar and femtomole quantities of carbohydrates in aqueous high-performance liquid chromatography. Its sensitivity is comparable to those of the most sensitive detection methods available for carbohydrate analysis. The atmospheric pressure argon glow discharge is sustained between two metallic electrodes and is operated at the output of a liquid chromatography system. The glow discharge induces oxidation in the eluting carbohydrates, forming acid(s) in amounts proportional to carbohydrate concentration. The relative acid concentration is monitored with a conductivity detector placed downstream from the discharge region. A detailed description of the detector and the parameters that control sensitivity is presented. The formation of hydrogen peroxide, hydronium ion, and UV-absorbing species in aqueous flowing glucose solutions is also studied using the glow discharge-conductivity detector. A comparison between these products and the products formed under similar conditions using high-energy radiation is discussed. PMID- 21639298 TI - Carbon disulfide measurement in marine waters using oxygen-doped electron capture detection with purge-trap preconcentration. AB - An oxygen-doped GC/ECD system has been found to be highly sensitive to carbon disulfide (CS(2)). The detector shows a linear response up to at least 13 pmol of CS(2) injected and has a detection limit of 30 fmol. By combining the oxygen doped GC/ECD with a purge-trap system, CS(2) in seawater can be readily separated and determined. A comparison is provided between an oxygen-doped ECD and a nondoped ECD. PMID- 21639299 TI - A (-)-menthyl bonded silica phase for chiral separations: synthesis and solid state NMR characterization. AB - A new (-)-menthyl bonded silica phase has been prepared by hydrosilation of a hydride silica intermediate. The hydride silica intermediate was synthesized by the reaction of a monoalkoxysilane (CH(3))(2)SiH(OEt) with silica gel, yielding a relatively high surface coverage (4.4 MUmol/m(2)) of SiH groups. This intermediate was then used successfully in the preparation of a monomeric (-) menthyl bonded silica phase. The bonded phase produced has been used for the chromatographic separation of enantiomers in a reversed phase mode (chiral separations). Solid state (13)C and (29)Si CP-MAS NMR spectroscopy and DRIFT spectroscopy provides valuable information on the structure of the different surface species formed on silica after modification. The surface coverage of the hydride silica intermediate and of the final bonded silica phase produced are also determined. It is found that this modification procedure can exclusively produce a monomeric coverage of SiH groups on the silica surface and can further produce a final monomeric bonded organic silica phase for the separation of enantiomers. PMID- 21639300 TI - Titration without Mixing or Dilution: Sequential Injection of Chemical Sensing Membranes. AB - A novel, miniaturized titration was developed using beads 35 MUm in diameter as semisolid aqueous titrant retained in a nonaqueous sample stream. Agarose beads with internally bound pH indicator served as a pH sensing membrane material swollen with aqueous NaOH titrant. The indicator monitored the remaining titrant within the agarose beads during perfusion with H(2)SO(4) in 1-butanol samples. Irreversible reaction of 2 mg bead layers was made possible by automated packing and disposal in a flow cell. This strategy substituted membrane advantages for the burdens of mixing and unnecessary dilution under laminar flow conditions. The agarose environment was conditioned with NaCl to tolerate dissolved salt in the sample. Transmittance measurements were made via fiber optics through FEP PTFE optical windows. A simple inverse relationship held between endpoint volume and acid concentration so that calibration curves were linear, R(2) = 0.9980. PMID- 21639301 TI - Thermospectrophotometric analysis of alkylamines utilizing ion association with tetrabromophenolphthalein ethyl ester. AB - Primary, secondary, and tertiary alkylamines react with tetrabromophenolphthalein ethyl ester dye (HTBPE) to form reddish charge transfer (CT) complexes in 1,2 dichloroethane (1,2-DCE). Absorption maxima of the complexes with primary amines occur at ~560 nm, with secondary amines at ~570 nm and tertiary ones at ~580 nm. CT complex formation constants between amines and HTBPE in 1,2-DCE at 25 degrees C increase in the order of primary, secondary, and tertiary amines but decrease with an increase in temperature. This phenomenon (thermochromism), which depends on the basicity of amines, was thermodynamically studied and used for the selective analysis of alkylamines. For example, tri-n-butylamine could be determined without interference from n-butylamine. PMID- 21639302 TI - An amperometric fructose biosensor based on fructose dehydrogenase immobilized in a membrane mimetic layer on gold. AB - A prototype amperometric fructose biosensor based on membrane-bound fructose dehydrogenase (Gluconobacter sp.) and the coenzyme ubiquinone-6 immobilized in a membrane mimetic layer on a gold electrode has been constructed and tested. A bare gold electrode first was modified through chemisorption of a mixture of octadecyl mercaptan and two short-chain disulfides, 3,3'-dithiodipropionic acid and cystamine dihydrochloride. The membrane-bound enzyme, coenzyme, and additional phospholipid were codeposited through a detergent dialysis protocol. The short-chain modifiers may provide electrostatic interactions with enzyme surface charges, while the alkanethiolate and phospholipids enable hydrophobic interaction with the largely lipophilic, membrane-bound enzyme. At oxidizing potentials, the enzyme electrode responded with catalytic current densities up to 45 MUA/cm(2) when exposed to fructose at 10 mM. The sensor exhibited a response time of less than 20 s, a sensitivity of 15 MUA/cm(2).mM and a detection limit of less than 10 MUM. Biosensor measurements of d-fructose in apple and orange juice agreed to within a few percent with those made with an enzymatic spectrophotometric assay. The membrane mimetic layer effectively blocked access of interfering ascorbic acid to the electrode surface. Only a 4% positive error was observed in the presence of ascorbic acid at 5% of the fructose concentration (2 mM), which indicates that this construct could be particularly useful for quantitation of fructose in citrus juice. PMID- 21639303 TI - Fast wave forms for pulsed electrochemical detection of glucose by incorporation of reductive desorption of oxidation products. AB - The electrochemical activity of Au electrodes held at constant potential for anodic detection of carbohydrates in alkaline media eventually decays to zero. This loss of response is a consequence of the accumulation of adsorbed oxidation products on the electrode surface. Although it is well-known that these "poisons" can be removed by oxidative desorption simultaneously with formation of surface oxide, we have discovered that electrodes fouled during the detection of glucose yield a cathodic peak at -0.77 V vs SCE resulting from reductive desorption of these species. Incorporation of the reductive desorption process into wave forms for pulsed electrochemical detection (PED) permits a significant decrease in the time periods traditionally allowed for the oxidative and reductive reactivation of the electrode with a resulting increase in wave form frequency. A 6.7-Hz wave form using E(red) = -1.00 V (t(red) = 10 ms), E(oxd) = +0.60 V (t(oxd) = 10 ms), and E(det) = +0.10 V (t(del) = 50 ms, t(int) = 50 ms) is applied for detection of glucose in a LC-PED system and is demonstrated to yield a sub-picomole detection limit with a linear dynamic range extending over three decades. PMID- 21639304 TI - Mechanism for the electrochemical stripping reduction of the nickel and cobalt dimethylglyoxime complexes. AB - Reductive coulometric stripping potentiometry, a technique not hitherto described, has been used to establish that the reduction of Ni(II) and Co(II) dimethylglyoximates, adsorbed on a mercury film electrode, is a 10-electron process. Exhaustive adsorption of Ni(II) or Co(II) complexes, in the 0-4 MUg L( )(1) concentration range, was achieved by vibrationally promoted electrolysis for 3 min of ~25 MUL volume samples, hanging under the working electrode in a nitrogen atmosphere. The adsorbed complexes were reduced by means of a constant current of 50 MUA. The technique was successfully used for the calibration-free determination of Ni(II) in certified seawater and river water reference samples. PMID- 21639305 TI - Application of a multivariate curve resolution procedure to the analysis of second-order melting data of synthetic and natural polynucleotides. AB - The melting behavior of several synthetic polynucleotides and a mixture of natural tRNAs was studied by monitoring the changes in the whole UV absorbance spectrum at several pH values. The second-order absorbance data were analyzed with a soft-modeling multivariate curve resolution procedure that allows the determination of the number of different species or conformations present along the melting experiment and the calculation of the melting profile and the pure spectrum for each chemical species or conformation. The melting temperature, T(m), for each thermal transition can be calculated from the melting profiles, and structural information on the different species or conformations can be obtained from their pure spectra. The multistranded species formed at certain pH conditions show several sharp thermal transitions related to the loss of the initial highly ordered structure. For these transitions, the mixture of species obtained in the denaturing process can be resolved when several data matrices, each giving additional information, are analyzed simultaneously with the mathematical procedure proposed. PMID- 21639306 TI - Enantiomeric Separation and Detection of 2-Arylpropionic Acids Derivatized with [(N,N-Dimethylamino)sulfonyl]benzofurazan Reagents on a Modified Cellulose Stationary Phase by High-Performance Liquid Chromatography. AB - (RS)-2-Arylpropionic acids (2-APAs) were derivatized with the fluorogenic reagents, 4-[(N,N-dimethylamino)sulfonyl]-7-piperazino-2,1,3-benzoxadiazole (DBD PZ) and 4-[[(N-hydrazinoformyl)methyl]-N-methyl]amino-7-[N,N (dimethylamino)sulfonyl]-2,1,3-benzoxadiazole (DBD-COHz), and their enantiomeric separation by a chiral stationary phase high-performance liquid chromatography was investigated in the reversed-phase mode with H(2)O/CH(3)CN or H(2)O/MeOH as the mobile phase on a column of cellulose tris(3,5-dimethylphenyl carbamate) coated on a silica gel support (Chiralcel OD-R). The derivatives with DBD-PZ were enantiomerically separated well under the elution condition of H(2)O/MeOH, based on the pi-pi interaction between the derivatives and the stationary phase. The rigid and bulky structure of DBD-PZ was demonstrated to be more effective as compared to the less rigid ones. The derivatives with DBD-COHz were more efficiently separated into each enantiomer with H(2)O/CH(3)CN as the eluent. The effective separation was based on hydrogen-bonding interaction between the acid hydrazide of the derivatives and the carbamoyl moiety of the stationary phase. There was a reversal in the elution order of the enantiomers between the two fluorescent derivatives. The detection limits obtained for each enantiomer were approximately 10-30 fmol on column. The derivatization with the reagent and the concomitant use of the reversed-phase and chiral stationary-phase HPLC were demonstrated to be useful for the enantiomeric quantification in rat plasma after intravenous administration of flurbiprofen racemate, a representative of 2-APAs. PMID- 21639307 TI - Multielemental chemical imaging using laser-induced breakdown spectrometry. AB - Multichannel laser-induced breakdown spectrometry (LIBS) is used to generate selective chemical images for silver, titanium, and carbon from silicon photovoltaic cells. A 2.5 mJ pulsed nitrogen laser and a spectrometer using charge-coupled device detection were employed. LIBS images were acquired sequentially by moving the sample located on a motorized x-y translational stage step by step while storing the multichannel LIBS spectrum for each position of the sample, followed by computer-based reconstruction of two-dimensional selective images from intensity profiles at several wavelengths. Depth distributions of carbon impurities are also reported. Room temperature and atmospheric pressure operation as used here remove the restrictions on sample size exhibited by other surface analysis techniques used for imaging applications. Thus, the sample size in LIBS imaging is in principle unlimited. A LIBS experiment does not require a sample to be conductive. Therefore, virtually all materials can be imaged. Although LIBS is a typical example of destructive analytical technique, multichannel detection as demonstrated here confers the possibility to LIBS of obtaining multielement information from a given surface area. Lateral resolution of 80 MUm and depth resolution of better than 13 nm were observed. The ultimate limitation to imaging the first layer of the surface in LIBS is the spectral signal-to-noise ratio as dictated by the ablation threshold of the material. PMID- 21639308 TI - Fourier transform infrared detection in miniaturized total analysis systems for sucrose analysis. AB - In this work, a flow system containing a micromachined lamella-type porous silicon reactor and a novel mid-IR fiber-optic flow cell were used for the enzymatic determination of sucrose in aqueous solution. The method relies on the enzymatic hydrolysis of sucrose to fructose and glucose catalyzed by beta fructosidase and on the acquisition of FT-IR spectra before and after complete reaction. beta-Fructosidase was covalently bound to the porous silicon surface of the channels in the microreactor. The porous silicon was achieved by anodization of the silicon reactor in a HF/ethanol mixture. For the measurement of small amounts of aqueous solution, a miniaturized flow cell was developed which consisted of two AgCl(x)Br(1)(-)(x) fiber tips (diameter, 0.75 mm) coaxially mounted in a PTFE block at a distance of 23 MUm. The flowing stream was directed through the gap of the two fiber tips which served to define the optical path length and to bring the focused mid-IR radiation to the place of measurement. Using this construction, a probed volume of ~10 nL was obtained. The calibration curve was linear between 10 and 100 mmol/L sucrose. Furthermore, the potential of this method was demonstrated by the analysis of binary sucrose/glucose mixtures showing no interference from glucose and by the successful determination of sucrose in real samples. PMID- 21639309 TI - Borohydride-Coupled HPLC-FPD Instrumentation and Its Use in the Determination of Dimethylsulfonium Compounds. AB - Novel HPLC instrumentation has been developed which employs an in-line sodium tetrahydroborate (borohydride) reaction step to generate volatile sulfur species from a variety of sulfonium compounds. Transfer of the resulting volatile sulfur containing products into the gas phase permits them to be monitored using sulfur specific flame photometric detection. The system has been evaluated for the determination of a collection of dimethylsulfonium compounds, comprising (dimethylsulfonio)propionate, S-methylmethionine (SMM), (dimethylsulfonio)-2 methylpropionate, dimethylsulfocholine, (dimethylsulfonio)acetate, (dimethylsulfonio)butanoate, and (dimethylsulfonio)pentanoate. Following their separation by either cation- or anion-exchange HPLC, these compounds react in line with the tetrahydroborate, generating dimethyl sulfide, which is then swept into a flame photometric detector. The development of chromatographic conditions for the resolution of the seven sulfonium compounds is described. In an example application of the instrumentation, the levels of SMM in parsley and cabbage were found to be 16 and 74 mmol kg(-)(1), respectively, on a fresh weight basis. PMID- 21639310 TI - Probing combinatorial library diversity by mass spectrometry. AB - The feasibility of a "massively parallel" mass spectrometric method for probing combinatorial library diversity is addressed theoretically for the example of computer-generated mass distributions of combinatorially synthesized peptide libraries containing between two and seven amino acids. We study the behavior of several "global" (integral) parameters of such mass distributions [Formula: see text] mass centroid, dispersion, skewness, and kurtosis. The centroid and dispersion are shown to carry information that may characterize the completeness of the synthetic effort. "Local" mass distribution parameters, e.g., "mass density" (number of peptides per mass interval), are also examined. The practical implementation and eventual limitations of such an approach are discussed as well. PMID- 21639311 TI - Laser Time-of-Flight Mass Spectrometry of PAH-Picrate Complexes. AB - The laser desorption/laser ionization time-of-flight (L2ToF), mass spectra of anthracene and the anthracene-picric acid charge transfer (C-T) complex have been compared at a desorption and ionization wavelength of 266 nm. Laser desorption/ionization spectra of anthracene were obtained at low temperatures ( 30 degrees C) to minimize the interference from gas phase ionization. Positive ion mass spectra of the picrate C-T complex at room temperature comprise the parent ion of anthracene and were devoid of signals associated with the picric acid component. The L2ToF analyses of a mixture of volatile and involatile EPA priority PAHs in picric acid show that low molecular weight PAHs form involatile charge transfer complexes. The present method reduces the possibility of volatile PAH loss during mass spectrometric analyses in vacuo. PMID- 21639312 TI - Direct current glow discharge mass spectrometry for elemental characterization of polymers. AB - A direct current glow discharge mass spectrometer has been used for a novel application, the sputtering and subsequent analysis of polymers. This was made possible by the application of a secondary cathode, a tantalum diaphragm placed in front of the nonconducting sample. Different types of polymers were measured (polytetrafluoroethylene, polycarbonate, and poly(vinyl chloride)). Important to note is that the mass spectra obtained are predominantly characterized as atomic, a major difference from the radio frequency GDMS spectra of polymers reported earlier. This facilitates quantitative elemental analysis for several reasons. PMID- 21639313 TI - Adsorption mechanism of trivalent metal ions on chelating resins containing iminodiacetic Acid groups with reference to selectivity. AB - The adsorption equilibria of seven trivalent metal ions (M(3+): Sc(3+), Y(3+), La(3+), Fe(3+), Al(3+), Ga(3+), and In(3+)) on chelating resins containing iminodiacetic acid groups (-LH(2)) were studied. Adsorption curves, measured under the conditions of metal ions in excess against chelating groups, directly indicated the metal-to-ligand ratio of the complexes formed in the resin phase. Iron and group 13 metal ions were adsorbed as (-L)(2)HM, while group 3 metal ions were adsorbed as (-L)(3)H(3)M and (-L)(2)HM. The adsorption constants for ( L)(2)HM found for all the metal ions were well correlated with the formation constants of iminodiacetate complexes in aqueous solutions. The actual adsorption of group 3 metal ions was significantly enhanced beyond that expected from this correlation because of the formation of (-L)(3)H(3)M. This is why the selectivity in the adsorption of trivalent metal ions differs from that in the complexation of iminodiacetate in aqueous solutions. The effects of anions and the number of iminodiacetic acid groups per unit weight of resins were also discussed. PMID- 21639314 TI - Measurement of the binding of proteins to polyelectrolytes by frontal analysis continuous capillary electrophoresis. AB - We have developed a novel technique, frontal analysis continuous capillary electrophoresis (FACCE), to study the binding of proteins to polyelectrolytes. Compared with existing electrophoresis methods such as conventional frontal analysis and the Hummel-Dreyer method, FACCE offers enhanced lower detection limits and is free from effects due to slow binding kinetics, thus making it suitable for studying equilibrium systems. In addition, with a single calibration, FACCE provides for efficient quantitative analysis. Here we report results obtained with beta-lactoglobulin as the ligand and sodium poly(styrenesulfonate) (NaPSS) as the ligand-binding substrate. For this model system, FACCE yields reproducible calibration curves and binding isotherms. The binding parameters so determined are compared with previous results for other protein-polyelectrolyte systems. PMID- 21639315 TI - Direct on-line injection in capillary electrophoresis. AB - A direct method of sample introduction for capillary electrophoretic techniques is described using a cross configuration and high-voltage shunting. No physical disturbance of the separation capillary inlet is required, and the feasibility of direct on-line injection is demonstrated. Both full- and pinched-mode injections are evaluated, with pinched-mode injections showing superior performance. In the pinched mode, only a portion of the cross volume is introduced onto the separation capillary, as a result, a lower volume is injected, and wall effects within the cross are minimized. Preliminary studies indicate a peak height reproducibility for replicate injections of better than 4.1%, with area reproducibilities of less than 3.1% for nonoverlapping solutes. Utilizing this direct on-line injection method, many rigid or restricted capillary geometries can be accommodated, and extension to the wide range of capillary electrophoretic techniques is feasible. PMID- 21639316 TI - Parametric modulation in liquid chromatography: multivariate optimization of mobile phase composition and temperature. AB - The concept and theory of parametric modulation are presented, and the strategy is demonstrated for multivariate optimization of mobile phase composition and temperature in liquid chromatography. Because each parameter to be optimized is maintained in separate and distinct zones along the column, the solutes are able to interact independently within each environment. Under these conditions, solute retention is a simple and rigorously predictable summation of the retention in each environment. Hence, parametric modulation is more accurate and requires fewer preliminary experiments than traditional optimization methods. This approach is demonstrated by application to the separation of isomeric polynuclear aromatic hydrocarbons using a polymeric octadecylsilica stationary phase, with methanol and acetonitrile mobile phases at temperatures from 23 to 45 degrees C. PMID- 21639317 TI - Effect of carbon loading on the extraction properties of C-18 bonded silica used for solid-phase extraction of acidic and basic analytes. AB - A range of C-18 bonded phases for solid-phase extraction (SPE), with different carbon loadings and degrees of end capping, have been evaluated for the extraction of model acidic and basic analytes (anisic acid and propranolol, respectively). High carbon loadings (18 and 22%) were associated with poor extraction properties with losses on application and wash steps. Recovery of extracted analytes from such materials was also inefficient, suggesting poor mass transfer properties. A low-carbon-loaded phase (5%), though capable of efficient extraction of the analytes, was seen to be poorly retentive. The best SPE phases for either analyte were those of intermediate carbon loading (~15%). Significant effects of end capping were only observed for the basic analyte propranolol, where retention was probably mediated by silanophilic interactions. PMID- 21639318 TI - Critical peak resolution in multicomponent chromatograms. AB - The critical degree of peak overlap is determined for multicomponent chromatograms. As the heights of adjacent peaks significantly differ, no single value of critical peak resolution can be used in multicomponent separations; one has to determine the distribution of the critical peak resolutions, taking into account the peak height distribution. An analytical expression is derived for the critical peak resolution, from which one can obtain the average or maximum probable peak resolution by integration. The theoretical model can be applied to any peak height distribution with ease. PMID- 21639319 TI - Analysis of fission products using capillary electrophoresis with on-line radioactivity detection. AB - Capillary electrophoresis has been used to separate metal ions characteristically associated with nuclear fission. Indirect UV absorbance and on-line radioactivity detection were used simultaneously to monitor the analytes. The radioactivity detector consists of conical plastic scintillating material with the capillary passing through the center to provide a 4pi detection geometry. The wide end of the cone is optically coupled to a photomultiplier tube. Transient isotachophoretic techniques were employed to stack large volumes of samples which had low specific activities. Radioactivity detection of (152)Eu and (137)Cs was achieved at the nanocurie level for 80-100 nL injections. The detector is approximately 80% efficient, enabling samples resident in the detector window for 0.1 min to be reliably assayed. The separation of (137)Cs and (137m)Ba isotopes, which are in secular equilibrium, was modeled to demonstrate the effects of the rapid decay of (137m)Ba. PMID- 21639320 TI - Selective Liquid Membrane Transport of Lead(II) by an Acyclic Polyether Dicarboxylic Acid Ionophore. AB - The effects of the chain structure and substitutents in six acyclic polyether dicarboxylic acids and one acyclic polyether carboxylic acid upon the efficiency and selectivity of pH-driven Pb(2+) transport in a bulk chloroform membrane system have been assessed. Among the carriers, 1,2-bis[2-(o carboxyphenoxy)ethoxy]-4-tert-butylbenzene (1) is found to exhibit high selectivity for transport of Pb(2+) compared with alkali metal cations and a variety of other divalent metal ion species. Ionophore 1 also extracts Pb(2+) from aqueous solution into chloroform with the loss of two protons. A 1:1 complex of Pb(2+) with di-ionized 1 was isolated. PMID- 21639321 TI - Picomolar assay of native proteins by capillary electrophoresis precolumn labeling, submicellar separation, and laser-induced fluorescence detection. AB - We report a method for the assay of proteins at concentrations lower than 10( )(10) M with as little as 200 amol of protein. High sensitivity is accomplished by derivatizing the epsilon-amino group of the protein's lysine residues with the fluorogenic dye 5-furoylquinoline-3-carboxaldehyde and use of a sheath flow cuvette fluorescence detector. Most proteins have a large number of lysine residues; therefore, a large number of fluorescent molecules can be attached to each protein molecule. In general, precolumn labeling improves sensitivity but degrades resolution due to the inhomogeneity of the reaction products from multiple labeling. However, we demonstrate that, through careful manipulation of the separation and reaction conditions, high sensitivity can be obtained without excessive loss in separation efficiency. Over 190 000 theoretical plates are obtained for fluorescently labeled ovalbumin. PMID- 21639322 TI - Effect of crown ethers on the ion-exchange behavior of alkaline Earth metals. Toward improved ion-exchange methods for the separation and preconcentration of radium. AB - The effect of selected crown ethers on the uptake of alkaline earth cations by sulfonic acid and diphosphonic acid-based cation-exchange resins from hydrochloric acid media is examined. The effect observed is shown to vary with the hydrophobicity of the crown ether. Water-soluble crown ethers enhance the sorption of certain cations, thereby improving the selectivity of the resin for other alkaline earths over calcium ion, an apparent result of a synergistic interaction between crown ether present in the resin phase and the ionic functional groups of the resin. In the presence of more hydrophobic crown ethers, a decrease in cation sorption is often observed, a result of the exclusion of the crown ether from the resin phase and the formation of cation-crown complexes in the solution phase. The result can be a reversal of the selectivity sequence ordinarily exhibited by the resin. PMID- 21639323 TI - Optimization of Chemiluminescent Nitrogen Detection for Packed-Column Supercritical Fluid Chromatography with Methanol-Modified CO(2). AB - A chemiluminescent nitrogen detector was optimized for packed-column supercritical fluid chromatography. Methanol modifier concentrations of 15% or less, an ozone flow of 5.8 mL/min, and a decompressed CO(2) flow between 240 and 310 mL/min were found to exhibit a maximum sensitivity of 5 ng for sulfamethazine (1 ng of N). The addition of a membrane drier to the "optimized" system further decreased the minimum detectable quantity (MDQ) to 0.5 ng (0.1 ng of N). In addition, by using a microbore column (2 mm i.d.) instead of an analytical scale column, the postcolumn decompressed flow split could be eliminated, further reducing the MDQ to 0.125 ng of sulfamethazine (0.025 ng of N). PMID- 21639324 TI - Radiochemical neutron activation analysis for trace chlorine in steels and alloys. AB - Chlorine is usually present at low concentrations in reactor materials and thermal neutron activation of (35)Cl produces (36)Cl, which has a long half-life and is a radionuclide of significance in nuclear waste disposal. This paper describes a radiochemical method that has been developed to measure low concentrations of Cl in reactor stainless steels, so that the amount of (36)Cl in radioactive wastes can be estimated. The method is based on the irradiation of a 1 g sample in a thermal neutron flux of 10(16) n m(-)(2) s(-)(1), followed by dissolution in HNO(3) with the addition of stable KCl carrier/tracer. The Cl is precipitated as AgCl, and the recovery is measured gravimetrically. The (38)Cl, which has a half-life of 37 min, is measured with gamma-ray spectrometry. The entire process, from irradiation to the end of counting, takes ~1.5 h. The recovery is near-quantitative, and the detection limit for Cl in most stainless steels is below 1 mg/kg. PMID- 21639325 TI - The application in processing analytical chemistry signals of a cardinal spline approach to wavelets. AB - The mth order B-spline N(m)(x) with integer knots generates a multiresolution analysis, ...?V(-1)?V(0)?..., with the mth order of approximation in wavelet transformation (WT). We have compared the WT method with Fourier transformation processing analytical chemistry signals in detail and have found that the WT method has many advantages. This method can directly provide the frequency domain distribution of a signal in a time domain. The algorithms are simpler and take less time in operation, and one needs only to decompose the digital signal rather than to undertake many transformations. Moreover, it is not necessary to preprocess the original signal to analyze the pattern of data and to know the statistical character of the noise. When the signal-to-noise ratio is 0.2, the processed results of theoretical and experimental data can still be satisfactory. PMID- 21639326 TI - Chiral discrimination in the gas phase using different transducers: thickness shear mode resonators and reflectometric interference spectroscopy. AB - The discrimination of optical isomers (enantiomers) in the gas phase has been performed using two different analytical tools: thickness shear mode resonators (TSMRs) and reflectometric interference spectroscopy (RIFS). The selective coatings included both enantiomers ((S)- and (R)-receptor) of a Chirasil-Val derivative (stationary phase material in GC) with octyl side chains. Successful discrimination of the enantiomers of different types of analytes (amino acids and lactates) was achieved. The results of both transduction methods were consistent and in good agreement with GC measurements. In addition, different mixtures of both enantiomers of the respective analyte were measured, and the enantiomeric composition could be quantitatively determined with excellent reliability. Since the sensors allow on-line monitoring (not possible with GC) of enantiomeric purity, an application in industrial synthesis (process control) of such compounds represents an interesting feature, especially with regard to the tested derivatives of lactic acid. PMID- 21639327 TI - Weighted least-squares approach to calculating limits of detection and quantification by modeling variability as a function of concentration. AB - The limit of detection and limit of quantification are current critical issues in environmental testing. In most laboratories, limits are currently calculated on the basis of the standard deviation of replicate analyses at a single concentration. However, since the standard deviation depends on concentration, these single-concentration techniques result in limits that are directly dependent on spiking concentration. A more rigorous approach uses a weighted least-squares regression analysis of replicates spiked at a series of concentrations [Formula: see text] a calibration design. In this work, the use of weighted tolerance intervals is introduced for estimating detection and quantification limits. In addition, models for estimating the weights used in calculating weighted prediction intervals and weighted tolerance intervals are presented. Using this method, detection and quantification limits were calculated for gas chromatographic analyses of 16 polychlorinated biphenyls. Results show that the approach developed provides improved estimates of analytical limits and that the single-concentration approaches currently in wide use are seriously flawed. Future work should reduce the data needed for the calibration design approach so that more rigorous detection and quantification limits can be routinely applied. PMID- 21639328 TI - Doped Thin-Film Sensors via a Sol-Gel Process for High-Acidity Determination. AB - An optical sensor has been developed for high-acidity ([H(+)] = 1-11 M) measurements. The sensor is made of thin films of silica sol-gels doped with an acid indicator. Acid- and base-catalyzed methods to make the sol-gel films have been studied, and the properties of the sol-gel sensors prepared by these methods are discussed. The acid-catalyzed method was found to give more robust films and has been optimized to prepare thin films which are mechanically and chemically stable for a period of at least 3 months. The performance of the sensors resulted in a relative standard deviation of less than 2.5%. The response time is short (1 s), and a small hysteresis was observed during reproducibility measurements with 2-10 M HCl solutions. PMID- 21639329 TI - The electroactive integrated optical waveguide: ultrasensitive spectroelectrochemistry of submonolayer adsorbates. AB - Highly sensitive spectroelectrochemistry of adsorbed films on ITO is demonstrated with the electroactive integrated optical waveguide (EA-IOW). The EA-IOW, a single-mode planar waveguide coated with an ITO layer, is ~10(4)-fold more sensitive to changes in absorbance occurring during electrochemical events versus a single-pass transmission spectroelectrochemical experiment, as demonstrated by reduction of surface-adsorbed methylene blue. Furthermore, the EA-IOW is selective to near-surface events, as it is relatively insensitive to absorbance by solutions of dissolved chromophores at <1 mM. The EA-IOW is also used to monitor the formation of Prussian Blue during the reduction of ferricyanide, an event that is not easily followed using current-detected cyclic voltammetry, due to interfering faradaic and non-faradaic electrochemical events. The optical background of the EA-IOW is potential-dependent and is explained by ion diffusion into the ITO and by voltage-dependent changes in optical constants for the material. Finally, the high sensitivity of the EA-IOW (relative to other evanescent-field-based spectroelectrochemical techniques) is discussed in terms of its design. PMID- 21639330 TI - A Poly(amphiphilic pyrrole)-Flavin Reductase Electrode for Amperometric Determination of Flavins. AB - The electropolymerization of a flavin reductase-amphiphilic pyrrole ammonium mixture previously adsorbed on the electrode surface provides the efficient entrapment of the enzyme in a functionalized polypyrrole film. The enzyme film was characterized and applied to the detection of several flavins and pyridine nucleotides. The response of the bioelectrode was based on the amperometric oxidation, at -0.1 V vs SCE, of the enzymically generated dihydroflavins in the presence of reduced pyridine nucleotides. The highest sensitivities and detection limits were 668 mA M(-1) cm(-2) and 4 nM for riboflavin, 179 mA M(-1) cm(-2) and 20 nM for flavin mononucleotide, and 107 mA M(-1) cm(-2) and 40 nM for flavin adenine dinucleotide. Direct electrochemical measurements at -0.65 V and electroenzymatic measurements at -0.1 V of riboflavin, carried out with these bioelectrodes, show that an amplification phenomenon occurs, due to an electrochemical recycling reaction of enzyme substrate. A nonamplified response to reduced pyridine nucleotides was also obtained in the presence of 0.1 mM riboflavin. The sensitivity and detection limit of the bioelectrode are 19.6 mA M(-1) cm(-2) and 0.2 MUM for NADH and 11.2 mA M(-1) cm(-2) and 0.5 MUM for NADPH. PMID- 21639331 TI - Diaphragm-free cell for trace determination of water based on the karl Fischer reaction using continuous coulometric titration. AB - A new type of diaphragm-free coulometric cell for continuous coulometric Karl Fischer titrations of water in the range 0.1-1000 MUg is described. The relative standard deviation obtained for titrations of 1 MUg amounts of water was typically 1%. The background due to diffusion of water from the air was normally in the range 0.3-0.9 MUg of water min(-1) depending on environmental humidity. The variation in the background was normally +/-0.01 MUg min(-1). The construction makes it possible, at any time in a sequence of titrations, to renew the catholyte by means of a Teflon plunger inside the cathode compartment. In this way, the interference effects caused by oxidizable reduction products of methyl sulfite which are formed at the cathode can be controlled in a very simple way. These products are rapidly eliminated by means of a normal titration before a new titration starts. The need for this draining step differs depending on the type of reagent used. The coulometric titration system makes use of true potentiometric end-point detection, and this principle makes it possible to control the iodine level at the end-point at much lower levels as compared with commercial instrumentation. The analytical advantages gained by this option are demonstrated for the determination of water in ethylenediamine, a task which was found to be impossible when using end-point concentrations in the range (3-7) * 10(-5) M, which is typical for the bipotentiometric indicating system used in commercial instruments. Recovery rates in the range 100-102% were obtained and are shown to be dependent on the type of reagent used. The most accurate results were obtained for an imidazole-buffered methanolic reagent in which the concentration of sulfur dioxide was kept relatively low (0.10 M). The diaphragm free cell described was shown to be compatible with all of the commercial reagents (designed for coulometry) investigated, including the well-known Hydranal products Coulomat A, AK, AG, AG-H, and AD. PMID- 21639332 TI - A triazine herbicide minisensor based on surface-stabilized bilayer lipid membranes. AB - This work describes an electrochemical technique that is suitable for rapid and sensitive screening of the triazine herbicides simazine, atrazine, and propazine. Egg phosphatidylcholine and dipalmitoylphosphatidic acid (DPPA) were used for the formation of self-assembled bilayer lipid membranes supported on silver wire (s BLMs). Evidence that BLMs could form on silver wires was collected by means of ellipsometry which was done to investigate samples consisting of lipids deposited on planar reflective silver films. The interactions of triazines with s-BLMs produced electrochemical ion current increases which reproducibly appeared within ~10 s after exposure of the lipid membranes to the herbicides. The sensitivity of the response was maximized by use of BLMs composed of 35% (w/w) DPPA and by alteration of the phase distribution within membranes by the introduction of 1.0 mM calcium ions in bulk solution. The mechanism of signal generation could be a result of rapid adsorption of the triazine on the surface of s-BLMs with a consequent rapid reorganization of the electrostatics of the membrane. The magnitude of the current signal was linearly related to the herbicide concentration, which could be determined at the nanomolar level. The present triazine minisensor exhibited good mechanical stability and longevity (routinely over 48 h), reproducible response characteristics (i.e., sensitivity and response to a given concentration of triazine in solution), fast response times, and low detection limits. The sensor can be simply and reliably fabricated at low cost. Studies have shown high selectivity for triazines in the presence of insecticides and pesticides. PMID- 21639333 TI - Extraction of elemental sulfur from an aqueous suspension for analysis by high performance liquid chromatography. AB - A bioreactor is being developed that produces elemental sulfur suspended in aqueous bioreactor contents. The concentration of elemental sulfur must be measured explicitly in order to study the efficiency of the conversion of sulfide to elemental sulfur. Extracting the sample with ethanol gave erroneous results when sulfide was present in solution. The extraction of aqueous elemental sulfur into petroleum ether prior to colorimetric determination was tested. When the aqueous matrix was simply deionized water, the extraction was poor. The development of a method of extraction of the sulfur into chloroform prior to quantification by high-performance liquid chromatography is described. The efficiency of the extraction was found to be greater than 90% in all matrixes tested and linear for aqueous elemental sulfur concentrations up to 200 mg/L. PMID- 21639334 TI - Thermal Stabilization of Enzymes Immobilized within Carbon Paste Electrodes. AB - In this note we report on the remarkable thermal stabilization of enzymes immobilized in carbon paste electrodes. Amperometric biosensors are shown for the first time to withstand a prolonged high-temperature (>50 degrees C) stress. Nearly full activity of glucose oxidase is retained over periods of up to 4 months of thermal stress at 60-80 degrees C. Dramatic improvements in the thermostability are observed for polyphenol oxidase, lactate oxidase, alcohol oxidase, horseradish peroxidase, and amino acid oxidase. Such resistance to heat induced denaturation is attributed to the conformational rigidity of these biocatalysts within the highly hydrophobic (mineral oil or silicone grease) pasting liquid. While no chemical stabilizer is needed for attaining such protective action, it appears that low humidity (i.e., low water content) is essential for minimizing the protein mobility. Besides their implications for electrochemical biosensors, such observations should lead to a new generation of thermoresistant enzyme reactors based on nonpolar semisolid supports. PMID- 21639335 TI - Dimethylcarbazoles in crude oils: comment on "liquid chromatographic separation schemes for pyrrole and pyridine nitrogen aromatic heterocycle fractions from crude oils suitable for rapid characterization of geochemical samples". PMID- 21639336 TI - Salicylate-selective electrode based on a biomimetic guanidinium ionophore. AB - A biomimetic strategy was employed in the development of oxoanion-selective ionophores containing the guanidinium functional group. These ionophores mimic the selective interaction observed between arginine residues of proteins and oxoanions. In previous work, it was demonstrated that a structurally rigid guanidinium ionophore exhibited excellent hydrogen sulfite selectivity (Anal. Chem. 1994, 66, 3188-3192). Herein, we describe guanidinium-containing ionophores that are selective for the oxoanion salicylate. The ability to rationally design anion-selective electrodes through this biomimetic strategy, and to both alter selectivity and improve response characteristics through structural changes to the ionophore, has been demonstrated. (1)H-NMR complexation and modeling studies were used to examine and correlate the selectivity observed with the structure of the guanidinium compounds. PMID- 21639337 TI - Infrared spectroscopy: instrumental factors affecting the long-term validity of chemometric models. AB - Data from instrumental techniques such as mid-infrared spectroscopy are increasingly being analyzed for sample identification and classification by chemometric methods based on principal component analysis (PCA). However, even modern spectrometers are subject to instability. This may affect PCA, because PCA selects the variables with the largest variance. This paper investigates the relative effects of sources of instrumental instability using a model developed for fruit puree classification. Single-beam spectra, potentially useful for on line analysis, saw their overall intensity decrease as the infrared source output and/or the detector sensitivity declined. Consequently, single-beam spectra were mainly differentiated by their overall intensity and had to be used with caution in the long term because this strongly affected the analyses. Absorbance spectra were not sensitive to source or detector decay but showed, in the long term, subtle band shape changes and frequency shifts. While these changes were not found to influence analyses involving very different samples, they diminished the success of analyses of data sets with small intrinsic variance. Where there was large spectral differences between sample classes, instrument-related factors were insignificant. However, where spectral differences were more subtle (with a single class), instrumental effects became more important. Suggestions are given to reduce the instrumental and experimental interferences on chemometric analyses, both when recording spectra and for managing spectral databases. PMID- 21639338 TI - Flow tube kinetics investigation of the mechanism of detection in the sulfur chemiluminescence detector. AB - Rate constants and their temperature dependencies were measured for the chemiluminescent reaction of ozone with the flame and furnace effluents of two commercially available sulfur detectors, the Sulfur Chemiluminescence detector (registered trademark of Sievers Instruments, Inc.) and the Chemiluminescent Sulfur Detector (registered trademark of Antek Instruments, Inc.). The absolute rate constants and activation energies agreed within experimental error with the well-established values for the SO + O(3) reaction in all detector configurations. When ozone was present in the reaction zone, the reaction of the furnace effluent with NO(2) also was in good agreement with the rate constant for the SO + NO(2) reaction. However, when ozone was not present in the reaction zone, little or no reaction occurred with NO(2). These results imply that an unknown sulfur species, X, is formed in the combustion zone and/or transfer line of the instrument and converted to SO upon reaction with ozone in the chemiluminescence detection cell. Based on our experimental results and known reaction rate constants, we can eliminate S(2), S(2)O, SO(2), S(2)O(2), HS, HSO, HSO(2), HOSO(2), and H(2)S as X. We hypothesize that X is S(3) formed in the association reaction of S atoms with S(2) within the transfer line. PMID- 21639339 TI - Analysis of Oligonucleotides by HPLC-Electrospray Ionization Mass Spectrometry. AB - A new interface procedure has been developed that allows, for the first time, the high-efficiency analysis of synthetic oligonucleotides up to 75 bases by reversed phase HPLC and on-line electrospray ionization mass spectrometry. For oligonucleotides up to 30 bases in length, single-base resolution can be obtained with low levels of cation adduct formation in the negative ion electrospray mass spectra. A key part of the method uses 1,1,1,3,3,3-hexafluoro-2-propanol as an additive to the HPLC mobile phase, adjusted to pH 7.0 with triethylamine. This novel additive results in both good HPLC separation and efficient electrospray ionization. The broad potential of this new method is demonstrated for synthetic homopolymers of thymidine (PolyT), fragments based on the pBR322 plasmid sequence, and phosphorothioate ester antisense oligonucleotides. This approach will be of particular utility for the characterization of DNA probes and PCR primers and quality control of antisense compounds such as phosphorothioates and their metabolites, as well as of materials used in clinical trials. PMID- 21639340 TI - Determination of Glucose in Submicroliter Samples by CE-LIF Using Precolumn or On Column Enzymatic Reactions. AB - Two enzymatic reactions combined with capillary electrophoresis (CE) are used to determine glucose contained in sample volumes of <=500 nL. In the first enzymatic reaction, glucose is oxidized in the presence of glucose oxidase producing hydrogen peroxide, which reacts quantitatively with the fluorogenic compound homovanillic acid catalyzed by the enzyme peroxidase. The second reaction generates a fluorescent species that is proportional to the glucose concentration. The reaction product is determined by CE using laser-induced fluorescence (LIF) as the detection mode. The overall reaction scheme is faster than commonly used precolumn derivatization procedures and can be performed using very small sample quantities. Alternatively, the enzymatic reactions can be performed on-column, similar to the electrophoretically mediated microanalysis approach, accommodating sample quantities in the nanoliter range. The on-column reaction is a simple and practical approach for the determination of glucose contained in low-volume samples by CE-LIF, where samples are injected directly into the capillary column without any pretreatment. However, sample handling and detectability of the precolumn approach proved to be superior. Determination of glucose using the precolumn and on-column reactions showed detection limits of 50 and 800 nM, respectively. The methods were shown to be linear in the range tested, 1-100 MUM and 100 nM-30 MUM, for the on-column and precolumn reactions, respectively. The reproducibility for each scheme was <5% RSD. To determine the possibility of using a noninvasive procedure for glucose monitoring, we used the CE-LIF methods to analyze human tear samples for glucose. The tear fluid samples were contained in a volume of ~200 nL. The concentration of glucose in the human tear samples analyzed using the precolumn and on-column procedures was ~138 MUM. PMID- 21639341 TI - Retention in field-flow fractionation with a moderate nonuniformity in the field force. AB - In some field-flow fractionation (FFF) techniques, the basic analyte-field interaction parameter, lambda, is not constant but varies within the channel cross section as a result of the nonuniformity of the force exerted by the field on the analyte. This is the case, for instance, in thermal FFF, because of the temperature dependence of the relevant physicochemical transport parameters. To account for this effect, a new FFF retention model is developed, allowing a linear variation of lambda from the accumulation to the depletion wall, which is assumed to describe correctly moderate nonuniformity in lambda in the vicinity of the accumulation wall. A methodology for sample characterization on the basis of this model is proposed. It associates lambda(app), the apparent lambda value derived from the retention ratio by means of the classical retention model, with a specific distance from the accumulation wall. An empirical relationship between that distance and lambda(app) is derived. In the high retention limit, it is found that this specific distance is not equal, as sometimes intuitively believed, to the mean distance of the molecule or particle cloud to the accumulation wall but is approximately equal to twice this mean distance. The validity of the proposed approach is checked. PMID- 21639342 TI - Peak shape distortions in the capillary electrophoretic separations of strong electrolytes when the background electrolyte contains two strong electrolyte co ions. AB - A series of 25 mM phosphate buffer background electrolytes were prepared from phosphoric acid and mixtures of lithium hydroxide and tetrabutylammonium hydroxide as pH adjusters and sources of background electrolyte co-ions. These background electrolytes were used for the capillary electrophoretic separation of quaternary ammonium analytes. Abnormally distorted peaks, different from the simple characteristic triangular peaks usually attributed to electromigration dispersion, were observed. In order to understand the origin of the greatly distorted peaks, capillary electrophoretic separations with two co-ion background electrolytes were numerically simulated using a mathematical model of the electrophoretic process. Generalized peak shape rules were derived from the simulations which can be used to predict the shape of the analyte, co-ions, and counterion concentration peaks, as well as the local electric field strength changes. Abnormal peak shape and peak disappearance can occur when the analyte peak and the noncomigrating system peaks overlap. PMID- 21639343 TI - Multiplex dsDNA Fragment Sizing Using Dimeric Intercalation Dyes and Capillary Array Electrophoresis: Ionic Effects on the Stability and Electrophoretic Mobility of DNA-Dye Complexes. AB - Methods have been developed for performing accurate, high-resolution, multiplex capillary electrophoresis separations of dsDNA using dimeric intercalation dyes as noncovalent labeling reagents. The quality of these separations is highly dependent on the cation present during electrophoresis. Using buffers that contain only one cation, we show that the tetrapentylammonium (NPe(4)(+)) ion results in high-resolution, high-sensitivity separations but that smaller ions such as sodium or the commonly used buffer ion tris produce low-resolution, low intensity separations of DNA-dye complexes. Using an 80 mM taps-NPe(4), 1 mM H(2)EDTA, pH 8.4, 0.8% HEC separation buffer, high-quality multiplex separations were performed using TOTO and buTOTIN, YOYO and TOED2, and TO and buTOTIN labeled restriction digests. In the taps-NPe(4) buffer, there is no significant mobility shift when complexes are formed with DNA-dye ratios from 100 to 5 bp per dye and very little dye transfer was observed. This property permits accurate multiplex sizing of samples having a wide concentration range simply by mixing the DNA with a dye solution before electrophoresis. This capability is demonstrated by diluting unpurified PCR products 10-, 100-, and 1,000-fold before mixing with a 1 nM TOTO solution and separating these samples with a PhiX174 HAEIII sizing ladder complexed with buTOTIN. Sizing precisions of better than 1% were obtained at all concentrations of target DNA. The mechanism for the increased DNA-dye complex stability and electrophoretic resolution in the taps-NPe(4) buffer is discussed. PMID- 21639344 TI - Capillary Ion Chromatography with On-Line High-Pressure Electrodialytic NaOH Eluent Production and Gradient Generation. AB - A small inexpensive system is described that allows high-performance suppressed anion chromatography on a capillary scale. A fully computer-controlled stepper motor-driven syringe-type dispenser, equipped with a 500 MUL-capacity glass syringe is capable of pumping at pressures up to 1000 psi when equipped with an appropriate inlet check valve. Fused-silica capillary columns ~50 cm in length and 180 MUm i.d., packed in-house with a commercial packing, provide excellent performance, significantly exceeding the efficiencies observed for the same packing in commercially available 2 mm bore format. The system operates with a pressure drop of <800 psi at a flow rate of 2 MUL/min. The system utilizes a novel electrodialytic NaOH eluent generator that is deployed on the high-pressure side of the pump and thus requires no special measures for electrolytic gas removal. This device permits both isocratic and gradient operation with excellent eluent purity; the NaOH concentration is generated linearly with applied current with near-Faradaic efficiency, up to a concentration of at least 100 mM. PMID- 21639345 TI - Statistical method for quantifying mobile phase selectivity in one- and two dimensional overpressured layer chromatography. AB - A method for selecting mobile phases for either one-dimensional (1-D) or two dimensional (2-D) planar chromatography is described and is applied to the separation of steroids by overpressured layer chromatography [Formula: see text] a form of forced-flow thin-layer chromatography [Formula: see text] using both normal- and reversed-phase chromatography. Two metrics are used for evaluating the separation quality of simulated chromatograms for each of 100 (or more) subsets of a set of 30 steroids in each of 15 1-D, and 105 2-D systems. The subsets vary in size between five and 25 steroids. Butyl acetate/toluene on silica gel and aqueous 2,2,2-trifluoroethanol are, on average, the highest and second highest ranked 1-D systems, respectively, for separating all subset sizes. These two systems are the constituent members of the system that is, on average, the highest ranked 2-D system for all subset sizes. The probability of the above systems being highest ranked decreases with decreasing subset size. There is only a 17% probability of butyl acetate/toluene on silica being the best system for separating a subset of five steroids, while there is a 3% probability of this being the worst system for this subset size. The spot capacity of each system can be estimated by considering 100 subsets of each size and noting the largest subset size that yields an acceptable value of one of the metrics used for measuring separation quality. The mobile phase selectivity may be quantified using the actual values of either of the two separation metrics, or by a nonparametric approach. The latter is used in such a way that a difference of unity in the ranking (for a given subset size) of two systems corresponds to a 95% probability that the higher ranked system will yield the better separation. PMID- 21639346 TI - Factors That Control Successful Entropically Driven Chiral Separations in SFC and HPLC. AB - With temperature increases, selectivity of chiral separations decreases until enantiomers coelute at an isoelution temperature. Above this temperature, elution order should reverse and selectivity will increase with temperature. In this region, separation is termed "entropically driven". Entropically driven chiral separations hold the promise of being able to concurrently increase selectivity and column efficiency by means of increased temperature. The ability to achieve such separations is hindered by high isoelution temperatures. The isoelution temperature is determined by a balance of enthalpic and entropic contributions. A variety of mobile phase modifiers are evaluated for their ability to moderate these contributions. Results suggest that more use should be made of non-alcohol modifiers. The major barrier to entropically driven separations was found to be the nonspecific retention increase that is characteristic when the critical temperature is traversed. Use of hexane in place of CO(2) shifts the position of the retention increase away from the temperature range used in this study, and dramatically successful entropically driven chiral separations are obtained. PMID- 21639347 TI - Direct On-Line Coupling of Capillary HPLC with (1)H NMR Spectroscopy for the Structural Determination of Retinyl Acetate Dimers: 2D NMR Spectroscopy in the Nanoliter Scale. AB - This paper presents the application of directly coupled capillary high performance liquid chromatography (capillary HPLC) and proton high-field nuclear magnetic resonance spectroscopy (NMR) for structural elucidation of a so-far unknown kitol isomer. One- and two-dimensional continuous- and stopped-flow NMR spectra were recorded in a 180 MUm i.d. capillary, corresponding to a detection volume of only 200 nL. Unequivocal structural assignment on the basis of 1D and 2D stopped-flow capillary HPLC-NMR experiments was performed. The kitol isomer mixture was present in a sample of thermally isomerized retinyl acetate and separated on a capillary column. PMID- 21639348 TI - Pesticide analysis with the pulsed-flame photometer detector and a direct sample introduction device. AB - New methods for fast, sensitive, and informative pesticide analysis in food products are described. These methods are based on sampling with a novel direct sample introduction device (DSI), gas chromatographic analysis, and pesticide detection with the pulsed flame photometric detector (PFPD). Sampling with the DSI is based on introduction of blended fruit or vegetable in a small glass vial that retains the harmful nonvolatile residue and is disposed of after the analysis. The DSI-GC-PFPD combination provides several new features that are demonstrated and discussed: (a) Extract-free analysis is achieved with the DSI, which serves as an effective alternative to sample extraction and cleanup methods. (b) Faster analysis is achieved through the use of the DSI due to the reduction of the upper GC program temperature, since the low-volatility compounds are retained in the sample vial. (c) Relatively efficient and uniform DSI thermal extraction and PFPD detection can be achieved, allowing the use of internal standards for pesticide calibration. (d) Higher sensitivity is provided by the PFPD as well as through the use of the DSI for sampling larger volumes of extract solutions. (e) Sulfur interference is eliminated with the PFPD in its phosphorus selective detection mode, using software exploiting differences in S and P delayed pulsed flame emission time dependence. (f) Sulfur pesticides are effectively analyzed by the PFPD in its sulfur-selective detection mode at the usual required levels and with less matrix interference than with NPD in the nitrogen mode. (g) Simultaneous sulfur and phosphorus pesticide analysis is demonstrated, including heteroatom identification and the provision of P and S atom ratio information in the analyzed pesticide. PMID- 21639349 TI - Stopless flow injection in asymmetrical flow field-flow fractionation using a frit inlet. AB - Stopless flow operation of asymmetrical flow field-flow fractionation (FFF) has been achieved by introducing a hydrodynamic relaxation method using a frit inlet. By using frit inlet injection, a focusing process which has been an essential part of runs at the asymmetrical flow FFF system can be completely avoided. Band broadening of an initial sample zone during hydrodynamic relaxation is discussed with equations related to the ratio of two inlet flow rates. For the successful achievement of particle relaxation and separation, it is necessary to apply a small ratio of sample inlet to frit inlet flow rate. Experimental results are reported for the evaluation of the system efficiency at various levels of hydrodynamic relaxation and for both normal and steric/hyperlayer modes of FFF runs using latex standards. Most importantly, it is shown that a high resolution and a high-speed separation of submicrometer-sized latex mixtures can be accomplished in asymmetrical flow FFF without using the conventional focusing relaxation process. PMID- 21639350 TI - QCM Operation in Liquids: Constant Sensitivity during Formation of Extended Protein Multilayers by Affinity. AB - The quartz crystal microbalance (QCM) is a well-established tool in mass sensitive detection. Due to recent improvements in experimental procedures, QCMs are finding increasing attention for applications in liquids. One important application is bioaffinity measurements for analytical or research purposes. The effect of the formation of solid films at a QCM surface, especially in gases or vacuum, is well understood. However, the situation is more complex in bioaffinity applications due to the comparably high viscosity of the liquid and the softness of the biological overlayer. Typically frequency responses found for protein layers exceed the values expected from simple models. The use of a hydrogel extending several hundred nanometers from the transducer surface as interacting matrix is common in bioaffinity applications and further increases complexity. Pure mass-related effects as well as viscosity-mediated effects may contribute to the overall frequency response observed experimentally. To improve our understanding of the effects during the formation of extended biological overlayers we have investigated systematically the formation of protein multilayers with a QCM in situ. The attenuation of the QCM oscillation by the liquid leads to a broadening of the resonance frequency. We have overcome this limitation by frequency-dependent admittance analysis and by curve fitting of the resulting admittance. A time resolution of 5 s and a noise of 0.2 Hz has been achieved with 6-MHz AT-cut quartz crystals operating in liquids. Protein multilayers were formed by successive incubations with a biotin-albumin conjugate and streptavidin. Frequency responses for dry protein layers in air were in agreement with mass changes estimated from the Sauerbrey equation. However, in water, the corresponding frequency decrease was increased by a factor of 4, thereby indicating that significant amounts of water are embedded in the hydrated protein layer. Unexpectedly a constant frequency decrease per layer was found during the successive formation of up to 20 protein layers (~400 nm). Neither noise nor drift increased with the number of protein layers. These results indicate that, despite the high hydration of the protein layers, viscosity induced effects play a negligible role and that the frequency decrease reflects primarily mass changes at the surface. PMID- 21639351 TI - Surface Plasmon Resonance Imaging Measurements of Electrostatic Biopolymer Adsorption onto Chemically Modified Gold Surfaces. AB - A combination of in situ and ex situ surface plasmon resonance (SPR) imaging experiments is used to characterize the differential electrostatic adsorption of proteins and synthetic polypeptides onto photopatterned monolayers at gold surfaces. The nonspecific electrostatic adsorption of proteins onto negatively charged self-assembled monolayers (SAMs) of 11-mercaptoundecanoic acid (MUA) is found to depend on the protein pI, solution ionic strength, and solution pH. The pH dependence of the electrostatic adsorption of the protein avidin onto a MUA SAM indicates that a full monolayer adsorbs at a solution pH greater than 5.0, and an "effective pK(a)" of 3.6 is determined for the avidin adsorption. This effective pK(a) is a combination of the pK(a) of the MUA monolayer and the ion pairing adsorption coefficient for the avidin. Additional SPR imaging experiments show that the electrostatic adsorption of the synthetic polypeptide poly-l-lysine (PL) onto a MUA SAM varies with molecular weight, forming a full PL monolayer for polypeptides with more than 67 lysine residues. PMID- 21639352 TI - Microfabricated electrochemical sensor for the detection of radiation-induced DNA damage. AB - An electrochemical biosensor protocol for the detection of radiation-induced DNA damage is described. The procedure employs a dsDNA-coated screen-printed electrode and relies on changes in the guanine-DNA oxidation signal upon exposure to ultraviolet radiation. The decreased signal is ascribed primarily to conformational changes in the DNA and to the photoconversion of the guanine-DNA moiety to a nonelectroactive monomeric base product. Factors influencing the response of these microfabricated DNA sensors, such as irradiation time, wavelength, and distance, are explored, and future prospects are discussed. Similar results are given for the use of bare strip electrodes in connection with irradiated DNA solutions. PMID- 21639353 TI - An Expansion for Quasi-Reversible Linear Potential Sweep Voltammetry and the Use of Euler's Transformation of Series. AB - An expansion for the current-voltage curves occurring in quasi-reversible linear potential sweep voltammetry is obtained in the form of an exponential (double) series. Euler's transformation of series is used for practical evaluation. Numerical precision problems limiting past applications of Euler's transformation can be overcome by the use of arbitrary precision arithmetic, now commonly available in mathematical software. Sample calculations are given, and the relation of the solution to previous work in the area is described. PMID- 21639354 TI - Laser-enhanced ionization of mercury atoms in an inert atmosphere with avalanche amplification of the signal. AB - A new method for laser-enhanced ionization detection of mercury atoms in an inert gas atmosphere is described. The method, which is based on the avalanche amplification of the signal resulting from the ionization from a selected Rydberg level reached by a three-step laser excitation of mercury vapor in a simple quartz cell, can be applied to the determination of this element in various matrices by the use of conventional cold atomization techniques. The overall (collisional + photo) ionization efficiency is investigated at different temperatures, and the avalanche amplification effect is reported for Ar and P-10 gases at atmospheric pressure. It is shown that the amplified signal is related to the number of charges produced in the laser-irradiated volume. Under amplifier noise-limited conditions, a detection limit of ~15 Hg atoms/laser pulse in the interaction region is estimated. PMID- 21639355 TI - Evanescent fiber-optic chemical sensor for monitoring volatile organic compounds in water. AB - The transport of trichloroethylene, 1,1,1-trichloroethane, and toluene in aqueous solutions through a polydimethylsiloxane film was modeled using a Fickian diffusion model to fit data obtained from an evanescent fiber-optic chemical sensor (EFOCS). The resultant diffusion coefficients for these analytes were respectively 3 * 10(-)(7), 5 * 10(-)(7), and 1 * 10(-)(7) cm(2)/s. Inclusion of an interfacial conductance term, defined as the ratio of the mass transport coefficient across the polymer surface and the analyte diffusion coefficient in the polymer, was required to accurately model the data. It was determined that the interfacial conductance terms were generally of the same order of magnitude for the analytes examined, suggesting a constant transport mechanism for the analytes. Linear chemometric algorithms were used to model the EFOCS response to aqueous mixtures of the three analytes with individual analyte concentrations between 20 and 300 ppm. Both partial least-squares and principal component regression algorithms performed comparably on the calibration sets, with cross validated root-mean-squared errors of prediction for trichloroethylene, 1,1,1 trichloroethane, and toluene of approximately 26, 29, and 22 ppm, respectively. The resultant prediction model was then used to determine analyte concentrations in an independent data set with comparable precision. PMID- 21639356 TI - The accuracy of instrumental neutron activation analysis of kilogram-size inhomogeneous samples. AB - The feasibility of quantitative instrumental neutron activation analysis (INAA) of samples in the kilogram range without internal standardization has been demonstrated by Overwater et al. (Anal. Chem. 1996, 68, 341). In their studies, however, they demonstrated only the agreement between the "corrected" gamma ray spectrum of homogeneous large samples and that of small samples of the same material. In this paper, the k(0) calibration of the IRI facilities for large samples is described, and, this time in terms of (trace) element concentrations, some of Overwater's results for homogeneous materials are presented again, as well as results obtained from inhomogeneous materials and subsamples thereof. It is concluded that large-sample INAA can be as accurate as ordinary INAA, even when applied to inhomogeneous materials. PMID- 21639357 TI - Laser Vaporization/Ionization Interface for Capillary Electrophoresis-Time-of Flight Mass Spectrometry. AB - Mass spectrometry (MS) is usually coupled on-line with capillary electrophoresis (CE) to analyze biomolecules by using electrospray ionization or continuous-flow fast-atom bombardment. We present a new design for laser vaporization/ionization time-of-flight mass spectrometry. CE, with its low flow rate (<1 MUL/min), is highly compatible with MS, even if the total column effluent is introduced directly. A UV laser is used to vaporize and ionize the solution eluting from the column. There is no need to have a makeup solvent. Using this system, we have analyzed a group of amines and peptides. The concentration detection limit of serotonin is in the 10(-)(7) M level. The separation and identification of an amine mixture by CE/MS demonstrates the complementary nature of the information. PMID- 21639358 TI - Electrostatic Interactions between Ru(bpy)(3)(2+) and Chromatographic Surfaces. AB - In HPLC, the zones of organic bases tail when silica-based stationary phases are used because the analytes and the surface are oppositely charged. In developing new stationary phases to achieve lower surface charge, a measure of surface charge is needed. The choice of a suitable analyte to quantitate electrostatic interactions is complicated by the acid-base equilibrium of the analyte itself and by nonelectrostatic interactions between the analyte and the surface, which alter the charge-induced tailing. This paper describes the study of the pH dependence of adsorption to isolate electrostatic interaction and the use of a cationic probe, tris(2,2'-bipyridine)ruthenium chloride (Ru(bpy)(3)(2+)), to sense surface charge without the complication of the probe's acid-base equilibria. The paper further describes the application of Gouy-Chapman theory to reveal the surface charge density. The results confirm that type A silica is considerably more acidic than type B silica and that horizontal polymerization makes type A silica perform as well as type B silica. PMID- 21639359 TI - A microscale biosensor for methane containing methanotrophic bacteria and an internal oxygen reservoir. AB - A microscale biosensor for continuous measurement of methane partial pressure based on a novel counterdiffusion principle is presented. Methane-oxidizing bacteria placed in the microsensor utilize oxygen from an internal oxygen reservoir when methane from the exterior diffuses through the tip membrane. The transducer is an internal oxygen microsensor with its tip positioned between the oxygen reservoir and the sensor tip membrane. The external partial pressure of methane determines the rate of bacterial oxygen consumption within the sensor, which in turn is reflected by the signal from the transducer. Tip diameters were down to 20 MUm, enabling us to study methane distribution on a microscale. The microscale construction also results in a low stirring sensitivity and a 95% response time down to 20 s. By tailoring the geometry, sensors can be made to exhibit a linear response in the full range of 0-1 atm partial pressure of methane or, alternatively, to exhibit a linear response only at lower concentrations, improving the sensitivity to below 0.1 kPa, corresponding to ~1 MUM in aqueous solution. Temperature, oxygen, and H(2)S interfere with the signal; no interferences were detected from H(2), NH(3), CO(2), or acetate. PMID- 21639360 TI - An Adaptation of Kubista's Method for Spectral Curve Deconvolution. AB - A chemometric approach to spectral curve deconvolution is described, evaluated, and applied to micellar systems. The technique is based on the method of principal component analysis of a spectral matrix followed by transformation of the abstract vectors into real spectra and concentrations. The approach reported here is similar to that of Kubista et al. (Anal. Chem. 1993, 65, 994-998). In the present study, however, more spectral information is known about the system of interest. This information is included in the deconvolution, which should, in general, increase the reliability of the method. From this method we obtain very reliable (noise-insensitive) lambda(max) values of indicator molecules in the micellar pseudophase free from contributions of the indicator in the aqueous phase. The water-to-micelle partition coefficients are also determined. The effects of noise and the extent of indicator partitioning on the reliability of the method are evaluated using model data. The application of the method to the study of eight indicators in a prototypical micellar system (sodium dodecyl sulfate) is presented. Extension of the method to other types of chemical studies such as the determination of kinetic rate constants and product spectra is briefly discussed. PMID- 21639361 TI - Determination of Uranium, Thorium, Plutonium, Americium, and Curium Ultratraces by Photon Electron Rejecting alpha Liquid Scintillation. AB - The determination of activities of thorium, uranium, plutonium, americium, and curium at very low levels has been performed by a new alpha liquid scintillation system (PERALS, name registered to Ordela, Inc.). The limit of detection has been determined for these nuclides with calculated values often lower than those obtained by other methods, like ICPMS/HP/Mistral, time-resolved laser-induced spectrofluorometry, and alpha spectrometry. All the results obtained show that the PERALS system is a promising method for the determination of these activities at very low levels. However, its energy resolution is inferior in comparison to that obtained by alpha spectrometry. For this reason, we have developed a process for separation of the five actinides as quickly and easily as possible. For each actinide, the conditions required to obtain optimal extraction yields and a complete separation have been determined. It is possible to perform the separation in only six extraction steps and to measure activities as low as a few millibecquerels per liter independently. This process has been applied with success to French granitic mineral or doped water and to complex media (biological samples like urines). In this latter case, the extraction recoveries are not quantitative, and it is necessary to determine the recovery yields by labeling with spikes like (230)Th, (232)U, (236)Pu, (248)Cm, and (148)Gd. PMID- 21639362 TI - Maximum likelihood multivariate calibration. AB - Two new approaches to multivariate calibration are described that, for the first time, allow information on measurement uncertainties to be included in the calibration process in a statistically meaningful way. The new methods, referred to as maximum likelihood principal components regression (MLPCR) and maximum likelihood latent root regression (MLLRR), are based on principles of maximum likelihood parameter estimation. MLPCR and MLLRR are generalizations of principal components regression (PCR), which has been widely used in chemistry, and latent root regression (LRR), which has been virtually ignored in this field. Both of the new methods are based on decomposition of the calibration data matrix by maximum likelihood principal component analysis (MLPCA), which has been recently described (Wentzell, P. D.; et al. J. Chemom., in press). By using estimates of the measurement error variance, MLPCR and MLLRR are able to extract the optimum amount of information from each measurement and, thereby, exhibit superior performance over conventional multivariate calibration methods such as PCR and partial least-squares regression (PLS) when there is a nonuniform error structure. The new techniques reduce to PCR and LRR when assumptions of uniform noise are valid. Comparisons of MLPCR, MLLRR, PCR, and PLS are carried out using simulated and experimental data sets consisting of three-component mixtures. In all cases of nonuniform errors examined, the predictive ability of the maximum likelihood methods is superior to that of PCR and PLS, with PLS performing somewhat better than PCR. MLLRR generally performed better than MLPCR, but in most cases the improvement was marginal. The differences between PCR and MLPCR are elucidated by examining the multivariate sensitivity of the two methods. PMID- 21639363 TI - Multielemental characterization of several brands of fullerenes and fullerene precursors by instrumental neutron activation analysis. AB - The characterization of the purity of fullerenes is based on the concept of defining the percentage amount of the main component (e.g., C(60), C(70), etc.) versus the minor fractions or microfractions of the homologue species and/or other polycyclic organic compounds. It has also to be considered that fullerene products may be contaminated in some degree by element impurities as well, both from the production processes and from the various precursor materials themselves. We report here for the first time detailed data on trace element impurities in various fullerene precursors as well as in the main C(60) and C(70) fullerene products of different producers, measured by instrumental neutron activation analysis. The concentration distributions of about 35 trace elements have been studied, including the determination of the relevant limits of detection. It could be established that all fullerene materials investigated contain a broad scale of trace elements distributed within a fairly wide concentration range, from ppb (ng/g) up to ppm (MUg/g) levels. For some impurity elements, extremely high concentration levels were found, indicating that elemental impurities have to be considered when studying the electrical, magnetic, and other features of the fullerenes. PMID- 21639364 TI - Fabrication and characterization of self-assembled spherical gold ultramicroelectrodes. AB - Spherical ultramicroelectrodes with diameters of 1-30 MUm have been prepared by self-assembly of Au nanoparticles and 1,9-nonanedithiol molecules at the tip end of glass micropipets. The electrodes were characterized by optical and scanning electron microscopy, cyclic voltammetry in aqueous and acetonitrile solution, and scanning electrochemical microscopy approach curves. A modified theory for hemispherical electrodes was used to compute the approach curves, which agreed with the experimental results. The construction strategy represents a bottom-up approach to the fabrication of microspherical electrodes. PMID- 21639365 TI - Multivariate curve resolution and trilinear decomposition methods in the analysis of stopped-flow kinetic data for binary amino Acid mixtures. AB - A stopped-flow method is proposed to carry out the kinetic development of the reaction between amino acids and 1,2-naphthoquinone-4-sulfonate by mixing analytes and reagent in a three-channel continuous-flow system. The process is monitored using a diode array spectrophotometer. Thus, every sample produces a data matrix built up from the spectra registered at regular steps of time. As the reaction is faster for secondary amino acids than for primary ones, it is possible to distinguish between the kinetic formation of their corresponding derivatives. The method is applied to the simultaneous determination of phenylalanine and proline by using second-order multivariate curve resolution. The derivatives of these two amino acids present some differences in both orders of measure, i.e., their spectra and kinetic profiles, which can be exploited advantageously to quantify one of the analytes in the presence of the other as interference, without including any information about this interference in the modeling of the system. PMID- 21639366 TI - Potentiometric response of conducting polymer electrodes for oxygen in neutral aqueous solutions. AB - The effect of oxygen on the potential of electrodes coated with a thin layer of various conducting polymers and immersed in aqueous neutral buffer solution has been studied. A theoretical equation describing the dependence of the potential on the concentration of oxygen has been derived and shown to be in conformity with the experimental results for polyaniline, polypyrrole, and poly(3 methylthiophene). The theory is based on the formation of charge-transfer complexes between oxygen molecule and the reduced (undoped) form of conducting polymer. PMID- 21639367 TI - Vibrational coupling as a probe of adsorption at different structural sites on a stepped single-crystal electrode. AB - Adsorption of carbon monoxide at step and terrace sites on a Pt(557) = Pt(s) [6(111) * (100)] electrode was detected with infrared spectroscopy. Vibrational coupling between adsorbates provided insights into the assembly of molecules at the different structural sites. The intermolecular coupling was weak at low coverages as CO ordered along the steps. For coverages between 40 and 70% of saturation, separate bands assignable to CO on steps and CO on terraces appeared. Coupling across this coverage range was markedly weaker on Pt(557) than on the structurally related Pt(335) = Pt(s)-[4(111) * (100)] electrode surface. The results indicate that, after the steps fill, CO populates the terraces on Pt(557) at random rather than by ordering in alignment with the steps. At coverages below saturation, vibrational bands assignable to CO molecules at step and terrace sites are affected differently by changes in electrode potential. The potential induced spectral changes for the terrace CO bands are similar to those of Pt(111)/CO, but the step CO bands show deviations from this trend at hydrogen adsorption potentials. PMID- 21639368 TI - On-line in vivo monitoring of endogenous quinones using microdialysis coupled with electrochemical detection. AB - The continuous on-line monitoring of endogenous quinones has been realized for the first time in an animal model of brain ischemia induced by a vasoconstrictor peptide, endothelin-1. A microdialysis probe, implanted in the striatum of a freely moving rat, was coupled on-line with an amperometric thin-layer cross-flow detector with a glassy carbon working electrode operating at -200 mV vs Ag/AgCl. The instrumental setup comprised a syringe pump pulse-damper consisting of an air bubble and a silica capillary, which permitted considerable reduction of background current fluctuations and allowed improved detection limits. This original configuration allowed the quantitation of micromolar amounts of total quinones, generated from dopamine during the reperfusion period, to be readily monitored. Several operational parameters have been investigated: flow rate, presence of oxygen in the perfusion fluid, and the working potential. The selectivity of the assay toward quinones was confirmed by studying possible interfering species such as ascorbate, hydrogen peroxide, riboflavin, and thiols. The results on freely moving rats have shown that the endogenous quinone amount was directly related to dopamine concentrations. The latter was determined by HPLC from dialysate samples collected at the outlet of the on-line system. HPLC studies showed that the primary quinone, generated from dopamine by bulk electrolysis, was also found in dialysates from ischemic brain. PMID- 21639369 TI - A Reference Electrode with Free-Diffusion Liquid Junction for Electrochemical Measurements under Changing Pressure Conditions. AB - A reference electrode for electrochemical measurements under changing pressure conditions is presented. The electrode of the Ag,AgCl|KCl type incorporates a mechanical pressure equalizing system to achieve independence from ambient pressure. An open liquid junction in connection with a controlled electrolyte outflow ensures accurate in situ measurements over periods of up to 10 days. Abrupt pressure changes (0.1 MPa s(-1)) effected short-time, reversible peak potential changes of less than 1.3 mV. The system was successfully used to monitor gastric pH in diving animals. The advantages of the new design are high tolerance to pressure changes, small size, no memory effects, low susceptibility to junction contaminations, and a long operating life, even under repeatedly changing pressure conditions. PMID- 21639370 TI - Electrochemical oxidation of cholesterol catalyzed by cholesterol oxidase with use of an artificial electron mediator. AB - Several redox dyes of phenazine and phenothiazine derivatives including 1-methoxy 5-methylphenazinium methosulfate (MPMS), thionin, azure A, and azure C work well as electron acceptors for reduced cholesterol oxidase (EC 1.1.3.6) (ChOx). Kinetic parameters and equilibrium constants of the enzymatic oxidation of cholesterol obtained suggest that the molecular size and/or hydrophobicity of the redox dyes determines their abilities as the electron acceptors. The electrochemical sensitivity to cholesterol in the detection system using ChOx and the above-mentioned redox compounds is determined by the electron acceptability of the redox compounds, and the employment of either MPMS or thionin allows electrochemical determination of cholesterol at a low electrode potential (0 V vs SCE) in the concentration range 0.25-2.5 mmol dm(-3), which covers that in normal human blood. PMID- 21639371 TI - Artificial neural network processing of stripping analysis responses for identifying and quantifying heavy metals in the presence of intermetallic compound formation. AB - Feed-forward neural networks have been trained to identify and quantify heavy metals in mixtures under conditions where there were significant complications due to intermetallic compound formation. The networks were shown to be capable of (i) correlating voltammetric responses with individual heavy metals in complex mixtures, (ii) determining the relationship between responses and concentrations (including nonlinear relationships due to overlapping peaks and intermetallic compound formation), and (iii) rapidly determining concentrations of individual components from mixtures once trained. Using simulated data, modeled after complex interactions experimentally observed in samples containing Cu and Zn, it has been demonstrated that networks containing two layers of neurons (a nonlinear hidden layer and a linear output layer) can be trained to calculate concentrations under a variety of complicated situations. These include, but are not limited to, cases where the response of the intermetallic compound formed is observed as a shoulder of one of the pure metals and cases where the response of the intermetallic compound formed is not observed in the potential window. In addition, the network described above was trained to simultaneously determine concentrations of four metals (Cu, Pb, Cd, and Zn) in a concentration range where all responses were complicated by intermetallic compound formation (1-500 ppb). PMID- 21639372 TI - Applications of sol-gel-derived membranes to neutral carrier-type ion-sensitive field-effect transistors. AB - Sol-gel-derived membranes encapsulating neutral carriers, such as valinomycin and a bis(crown ether) derivative, were fabricated as novel ion-sensing membranes for neutral carrier-type ion-sensitive field-effect transistors (ISFETs). The neutral carrier-type sol-gel-derived membranes obtained with an initial diethoxydimethylsilane/tetraethoxysilane ratio of 3 gave excellent results for membrane processibility and sensitivity, response time, and selectivity in the resulting ion sensors. The sol-gel-derived ion-sensing membranes containing neutral carriers are superior in the thrombogenic property to their corresponding plasticized poly(vinyl chloride) membranes. The potassium and sodium ISFETs thus obtained were successfully applied to the ion assay in blood sera. PMID- 21639373 TI - Spatial location of the space charge effect in individual ion clouds using monodisperse dried microparticulate injection with a twin quadrupole inductively coupled plasma mass spectrometer. AB - Pulses of analyte and matrix ions from individual drops are measured simultaneously using a twin quadrupole inductively coupled plasma mass spectrometer (ICP-MS). The sample solution is introduced by monodisperse dried microparticulate injection (MDMI). At modest Pb concentrations (500 ppm), a shoulder on the leading edge of the Li(+) signal appears. At higher matrix concentrations (1000 to at least 1500 ppm), a dip in the leading edge of the Li(+) signal develops. These changes in the shapes of the Li(+) pulses are attributed to space charge effects in the extraction system and ion optics of the mass spectrometer. A qualitative depiction for this behavior is proposed, in which the Li(+) ions are deflected out of the preferred ion path and then refocused by the ion optics. Part of the Li(+) ion cloud is driven ahead of the Pb(+) cloud, and part is trapped behind the Pb(+) cloud. The result is a shoulder on the leading edge of the Li(+) signal. With the Pb matrix present, the shapes of the analyte ion pulses are sensitive to the voltages applied to the first two ion lenses, especially the extractor lens. This observation shows that the part of the matrix effect that occurs in the ion optics takes place mainly in the first two lenses. PMID- 21639374 TI - Infrared Spectra Simulation of Substituted Benzene Derivatives on the Basis of a 3D Structure Representation. AB - The identification of chemical compounds from their infrared spectra faces new challenges from novel experimental techniques such as combinatorial chemistry. To rapidly provide estimates for the infrared spectra of candidate structures, an empirical approach to the modeling of the relationships between the 3D structure of a molecule and its infrared spectrum has been developed. This method is based on a novel 3D structure representation and a powerful modeling technique, a counterpropagation neural network. A dataset of 871 mono-, di-, and trisubstitued benzene derivatives is analyzed with this approach. PMID- 21639375 TI - Room-temperature phosphorescent palladium-porphine probe for DNA determination. AB - Room-temperature phosphorescence (RTP) enhancement of a palladium-porphine complex by DNA was studied. Studies involving calf thymus DNA and calf liver RNA revealed that the Pd-porphine complex bound to double-stranded DNA displayed intense RTP, while a very weak RTP emission was observed in the presence of equal molar concentrations of RNA. This fact founded a basis for selective determination of DNA in the presence of RNA. Maximum phosphorescence was observed at pH 7, with maximum excitation and emission wavelengths at 435 and 680 nm, respectively. Under optimal experimental conditions, the calibration graphs were linear up to 6 * 10(-)(5) MUM DNA. The detection limit was 5 * 10(-)(8) MUM and the relative standard deviation (10 replicates) +/-2.6% in the linear range. DNA extracted from human colon tissues could be successfully determined. PMID- 21639376 TI - Environmental applications of gas chromatography/atomic emission detection. AB - A gas chromatograph/atomic emission detector (GC/AED) system has been evaluated for its applicability to environmental analysis. Detection limits, elemental response factors, and regression analysis data were determined for 58 semivolatile environmental contaminants. Detection limits for injected analytes ranged from 0.17 to 3.0 ng on the hydrogen 486-nm channel, from 1.0 to 5.0 ng on the nitrogen 174-nm channel, from 0.65 to 11.7 ng on the oxygen 777-nm channel, from 0.071 to 3.0 ng on the chlorine 479-nm channel, and from 0.023 to 0.038 ng on the sulfur 181-nm channel. Mean elemental response factors (ERFs) measured on these channels, relative to the carbon 496-nm channel, were hydrogen, 0.084 (mean %RSD = 6.6); nitrogen, 0.246 (mean %RSD = 19); oxygen, 0.459 (mean %RSD = 16); and chlorine, 0.417 (mean %RSD = 3.6). The higher precision obtained for hydrogen and chlorine, relative to that for nitrogen and oxygen, is attributed to the ability to scan these elemental channels in the same GC run as the carbon 496-nm channel (diode array wavelength range limitation of ~40 nm/run). Mean ERFs of standard compounds were used to determine the molecular formulas of chlorinated hydrocarbons and chlorinated organosulfur compounds in a contaminated environmental soil sample. These formulas are in good agreement with the molecular weights and chlorine isotopic data obtained from low-resolution gas chromatography/mass spectrometry. PMID- 21639377 TI - Surface-enhanced Raman scattering from an etched polymer substrate. AB - An etched polymer was evaluated as a substrate suitable for routine SER analysis and was found to perform similarly to crossed gratings and posted quartz wafers. Silver particles were layered onto sheets of etched poly(ethylene terephthalate) by chemical reduction, sputtering, and vapor deposition. Enhancement factors ranged from 10(4) to >10(5) for crystal violet on chemically reduced and sputtered surfaces, respectively. Ranges of linearity up to 3 orders of magnitude were attained with limits of detection of ~5 pg. The variability of response was <20% RSD for both routinely and painstakingly prepared samples. To induce enhancements beyond those possible from the dry substrate, a thin film of water was held on the analyte-treated substrate surface in a liquid state. Typical spectra of nine analytes with amino, carboxylate, and/or nitro groups were collected from the most active substrate. PMID- 21639378 TI - Two-channel sequential single-molecule measurement. AB - Experimental results are given which demonstrate the sequential detection of single molecules with a measurement efficiency of near unity. IR140 dye molecules are detected in sequential probe volumes within a flowing stream through a 9 MUm i.d. capillary. The measurement of single molecules was confirmed by means of autocorrelation, photobleaching, visual observation, and cross-correlation analysis. The number of single molecules photobleached prior to being measured in the second probe region was in excellent agreement with the bulk studies described by a photodestruction curve. A cross-correlation peak with a temporal delay corresponding to the interprobe volume transit time and a width in agreement with parabolic flow give a clear indication of sequential detection. The near unity measurement efficiencies for both channels indicates the great potential for rapid quantitative analysis of dilute solutions. PMID- 21639379 TI - Trace level organics in hydrofluoric Acid determined by attenuated total internal reflection infrared spectroscopy. AB - Trace levels of organic impurities in the hydrofluoric acid solutions were measured by a multiple internal reflection infrared spectroscopic (MIRIS) technique. The MIRIS utilizes a clean attenuated total reflection (ATR) silicon crystal to extract organic impurities from the HF solutions. An open beam single channel background spectrum combined with statistical analysis was used to ensure the reproducibility of absorbed organics measurement. The hydrofluoric acid samples were analyzed under an ultrapure nitrogen blanket to avoid airborne organics interferences. The adsorbed organic contaminants were found to randomly orient on the silicon ATR probe surface. A higher level of organic impurities was found in the more concentrated hydrofluoric acid solutions. PMID- 21639380 TI - High-density, covalent attachment of DNA to silicon wafers for analysis by maldi tof mass spectrometry. AB - A method is described for the covalent attachment of DNA to a solid surface at high density for hybridization detection by mass spectrometry. A silicon wafer is functionalized to place an amino group on the surface; a heterobifunctional cross linking agent is then reacted with the primary amine to incorporate an iodoacetamido group. An oligodeoxynucleotide containing a 3'- or a 5'-disulfide is treated with a reducing agent, resulting in a terminal free thiol, which is then coupled to the iodoacetamido surface. Analysis of the surface reveals that the amount of covalently bound oligodeoxynucleotide is 250 fmol of DNA/mm(2) with ~40% of the immobilized oligodeoxynucleotides available for hybridization. Matrix assisted laser desorption/ionization time-of-flight mass spectrometric (MALDI-TOF MS) analysis reveals that the covalent linkage to the support remains intact, only the annealed strand is desorbed by the laser, and the amount of DNA hybridized to the array is sufficient for detection. PMID- 21639381 TI - Determination of iridium in natural waters by clean chemical extraction and negative thermal ionization mass spectrometry. AB - Methods for the precise, routine measurement of Ir in seawater, riverwater, and estuarine water using isotope dilution negative thermal ionization mass spectrometry (ID-NTIMS) have been developed. After equilibration with a (191)Ir enriched spike, Ir is separated from solution by coprecipitation with ferric hydroxide, followed by anion exchange chromatography using a reductive elution technique. UV irradiation is employed for the decomposition of trace organics, which interfere with negative ion production. IrO(2)(-) ions are produced in the mass spectrometer by heating the sample on a Ni-wire filament in the presence of Ba(OH)(2). Detection efficiencies ranged from 0.1% to 0.3%. We have used these procedures to determine the concentrations of Ir in 4 kg samples from the Pacific Ocean, the Atlantic Ocean, the Baltic Sea, and the rivers supplying the Baltic. Our chemical procedures introduce a total blank of ~2 * 10(8) atoms per sample. The distribution of Ir in the oceans is fairly uniform, averaging ~4 * 10(8) atoms kg(-)(1). The concentrations in the rivers supplying the Baltic Sea range from (17.4 +/- 0.9) * 10(8) for a pristine river to (92.9 +/- 2.2) * 10(8) atoms kg(-)(1) for a polluted river. The distribution, speciation, and transport of Ir in natural waters can now be subjected to intensive study. PMID- 21639382 TI - Low Blank Preconcentration Technique for the Determination of Lead, Copper, and Cadmium in Small-Volume Seawater Samples by Isotope Dilution ICPMS. AB - A simple low-blank method is described for the analysis of Pb, Cu, and Cd in seawater using Mg(OH)(2) coprecipitation and isotope dilution inductively coupled plasma mass spectrometry (ICPMS). Here, 20-40 MUL of 9 M aqueous NH(3) is mixed into a 1.3 mL seawater sample spiked with enriched isotopes of Pb, Cu, and Cd. After centrifugation, the supernatant is discarded and the Mg(OH)(2) precipitate dissolved in 100 MUL of 5% HNO(3) for ICPMS analysis. This method is simple, accurate, and precise, with detection limits of Pb = 1.3 pM, Cu = 39 pM, and Cd = 5.0 pM and blanks of Pb = 0.62 pM, Cu = 27 pM, and Cd = 6.0 pM. The method is demonstrated by oceanographically consistent profiles of these trace metals at an ocean station in the eastern North Atlantic. PMID- 21639383 TI - On-line linear sweep voltammetry-electrospray mass spectrometry. AB - A three-electrode electrochemical cell coupled to an electrospray (ES) mass spectrometer has been developed and employed for on-line linear sweep voltammetry ES mass spectrometry and electrochemical reaction studies. Generated in this type of electrochemistry/mass spectrometry (EC/MS) experiment are ion abundance vs potential profiles that provide molecular weight and structural information pertaining to intermediates and products of anodic oxidation of diphenyl sulfide, anodic pyridation of 9,10-diphenylanthracene, and reduction of nitrobenzene. Anodic oxidation of diphenyl sulfide in methylene chloride media at the platinum electrode yielded diphenyl sulfoxide and pseudodimer sulfonium ion along with products of further oxidation. The obtained ion abundance vs potential profiles offer additional evidence to corroborate previously proposed reaction mechanisms for pseudodimer sulfonium ion generation from diphenyl sulfide, as well as pyridine addition to DPA radical cation also in methylene chloride at the platinum electrode. From the latter experiment, a newly found reaction product, [DPA(py) - H](+), is reported. Despite the inherent tendency of ES devices to induce oxidation processes in the positive ion mode, intermediates and products of nitrobenzene reduction in nonaqueous and organic/water media were detected in both negative and positive ES modes. PMID- 21639384 TI - Mass spectrometric inverse gas chromatography: investigation of polymeric phase transitions. AB - An improved inverse gas chromatographic method involving the use of a mass specific detector for the determination of the glass transition temperature of polymeric materials is described. The new method allows the use of several probe solutes simultaneously with an automated, closed-loop injector and stepped temperature programming. The result is a single continuous chromatogram for each probe solute over a range of temperatures encompassing the glass transition temperature, T(g). Several different methods for the exact determination of T(g) from the chromatogram were investigated, including the classical van't Hoff-type plots with retention volumes calculated from both the peak maximum and first moment values of the elution peaks. Two new methods are also proposed for the evaluation of T(g) from either the temperature dependence of the second moments of the elution peaks for probe solutes or simple inspection of the variation of elution peak height (width) with temperature. All four methods for the determination of T(g) are evaluated with three probe solutes and four different polymers, viz., poly(methyl methacrylate), poly(ethylene terephthalate), polycarbonate, and two batches of polystyrene with different molecular weights and T(g) values. Three phenomenological models were used to interpret the chromatographic retention mechanisms of the solute probes in glassy and rubbery polymers. These are (i) the classical adsorption/absorption model for glass and rubber polymers, (ii) the single absorption mechanism model, and (iii) a dual mode model previously used to explain the sorption of gases, such as CO(2), in glassy polymers. It is concluded that no single approach is adequate to interpret the experimental results for all of the systems, although each model is adequate for some individual solute/polymer combinations. PMID- 21639385 TI - Tandem fourier transform mass spectrometry studies of surface-induced dissociation of benzene monomer and dimer ions on a self-assembled fluorinated alkanethiolate monolayer surface. AB - A new instrument configuration based on a Finnigan FTMS-2000 platform has been applied to the study of surface-induced dissociation (SID) in this research. Benzene monomer ions C(6)H(6)(+) and dimer ions (C(6)H(6))(2)(+) were impacted on a fluorinated self-assembled monolayer surface at collision energies ranging from 1 to 70 eV. Benzene cations were chosen for this study because the fragmentation characteristics of the molecular cation are well known and its SID has been thoroughly investigated. SID spectra obtained by FTMS-SID are very similar to those reported in the literature for the same surface but exhibit much higher mass resolution. A comparison study of collision-induced dissociation (CID) and SID of benzene molecular cations was performed utilizing the same ICR cell and ion detection protocol. It is demonstrated that SID provides both much higher energy deposition and a narrower internal energy distribution than CID. The present instrument geometry and experimental protocol demonstrate much higher efficiencies than previous SID studies by FTMS and much higher mass resolution than previous SID studies using other types of mass analyzers. PMID- 21639386 TI - Characterizing oligosaccharides using injected-ion mobility/mass spectrometry. AB - Injected-ion mobility/mass spectrometry techniques have been used to measure the reduced ion mobilities for negatively charged raffinose, melezitose and alpha-, beta-, and gamma-cyclodextrins formed by electrospray ionization. At low injection energies, the mass spectra are dominated by negatively charged (deprotonated) parent ions. At high injection energies, the mass spectra recorded for the cyclodextrins and raffinose display peaks that result from cross-ring cleavage of individual sugar units. Melezitose dissociates by cleavage of the glycosidic bonds. The ion mobility distributions can be used to distinguish between different isomeric forms of parent and fragment ions having the same mass to-charge ratios. PMID- 21639387 TI - Pulsed UV Laser-Induced Stationary Capillary Vibration for Highly Sensitive and Direct Detection of Capillary Electrophoresis. AB - A stationary wave of the capillary vibration effect was successfully induced by a series of short laser pulses. This wave could be applied to highly sensitive detection of capillary electrophoresis as well as the already reported capillary vibration induced by an intensity-modulated CW laser (CVL effect). Generally, pulses with much shorter width than the period of the natural frequency of the vibrating system cannot induce a standing vibration. However, utilizing the time constant of CVL determined by heat dissipation time, we found conditions which could induce a stable stationary wave of the capillary by a series of nanosecond light pulses. We used the KrF excimer laser operated at 248 nm with a pulse width of 60 ns and output of ~10 MUJ/pulse as the CVL excitation source and applied it to highly sensitive detection of nonderivatized amino acids at the femtomole level. The sensitivity was at least 2 orders of magnitude superior to that of a commercially available UV absorbance detector. This technique extends the CVL's spectral regions. For example, in the UV region, where many biological materials have significant absorption bands, this technique will extend analytical applications in capillary electrophoresis by eliminating the need for a derivatization process. PMID- 21639388 TI - High-efficiency solvating gas chromatography using packed capillaries. AB - In this study, column efficiency in packed capillary column solvating gas chromatography (SGC) was investigated. Long (>3 m) fused silica capillaries with an inner diameter of 250 MUm were packed with 10 and 15 MUm spherical porous (300 A) octadecyl bonded silica particles using a CO(2) slurry packing method. A 336 cm * 250 MUm i.d. fused silica capillary containing 10 MUm particles provided a total column efficiency of 264 000 plates (k = 0.41), corresponding to a reduced plate height of 1.27, using CO(2) as the mobile phase at a column inlet pressure of 260 atm. A minimum plate height of 12.7 MUm and a maximum plate number per unit time of 813 plates/s were obtained using packed capillary SGC. Retention factors were dependent on the column inlet pressure but independent of the pressure gradient along the column. Gasoline and diesel samples were separated under SGC conditions, and the results were comparable to those obtained using typical open tubular column gas chromatography. PMID- 21639389 TI - Analysis of corticosteroids in biofluids by capillary electrochromatography with gradient elution. AB - Capillary electrochromatography (CEC) with gradient elution was used to separate mixtures of corticosteroids (adrenosterone, hydrocortisone, dexamethasone, fluocortolone) in extracts of equine urine and plasma. Urine samples were first purified using solid phase extraction. Two purification steps were necessary to prevent contamination of the CEC column. Plasma was purified using automated dialysis. A laboratory-built CEC interface, connected to a gradient HPLC system, delivered samples and mobile phase to the CEC column. CEC was performed in fused silica capillaries of 50 MUm i.d., 24 cm total length, and 16 cm effective length packed with Apex ODS, 3 MUm particle size. The mobile phase was ammonium acetate (5 mM) in water/acetonitrile. Acetonitrile in the mobile phase was varied from 9 to 80% (v/v) using the gradient HPLC system. Detection was by UV absorbance at 240 nm. Samples, 10-250 MUL, were injected into the mobile phase stream and loaded onto the CEC column under an applied field of 1.04 kV cm(-1) and a CEC column head pressure of 12 bar. Mobile phase flow rate through the sampling interface was 100 MUL min(-1). The system was reproducible and could be left in unattended operation for long periods. After injection of 200 urine extracts, a broadening of peaks was observed but the CEC column was still serviceable. PMID- 21639390 TI - Simultaneous Determination of Ionization Constants and Isoelectric Points of 12 Hydroxy-s-Triazines by Capillary Zone Electrophoresis and Capillary Isoelectric Focusing. AB - Capillary zone electrophoresis (CZE) was used to separate and determine simultaneously the pK(1), pK(2), and pI values of 12 environmentally relevant hydroxytriazines (hydroxymetabolites of atrazine, terbuthylazine, simazine, and propazine and four (arylamino)-s-triazines) and observe the effects of the alkylamino and arylamino substituents on the measured values. Capillary isoelectric focusing (CIEF) methods were developed to measure the pI of these compounds and compare those values with the CZE-measured pI's. CZE and CIEF can provide accurate pK and pI values reasonably fast, and pI values measured by the two techniques agree. Knowledge of the pK and pI values of hydroxytriazines is important for an understanding of the binding mechanisms of these molecules in environmental matrices. Because the hydroxytriazines may exist as a myriad of species [Formula: see text] neutral, charged, zwitterionic, and keto-enol tautomeric [Formula: see text] depending on structure and pH, we briefly addressed the existence of these species relative to their electrophoretic analysis. PMID- 21639391 TI - Effect of acetonitrile/water mobile-phase composition on adsorption characteristics of reversed-phase liquid chromatography. AB - The influence of volumetric composition of acetonitrile in a mobile phase on adsorption characteristics of reversed-phase liquid chromatography using octadecylsilyl (ODS)-modified silica gel was studied by the pulse response method and the moment analysis. The results were compared with those obtained for the reversed-phase system consisting of ODS-silica gel and methanol/water mixtures. In both systems, surface diffusion was dominant for intraparticle diffusion in ODS-silica gel particles. The contributions of three mass transfer steps in a column to peak broadening were of about the same order of magnitude. The activation energy of surface diffusion, E(s), was found to be larger than the isosteric heat of adsorption, Q(st). Because similar tendencies were observed for these adsorption characteristics, adsorption mechanisms may be analogous in both chromatographic systems. However, absolute values of the adsorption equilibrium constant, K, the decreasing ratio of hydrophobic surface area, DeltaA/A, Q(st), and E(s) for acetonitrile/water systems were smaller than the corresponding values for methanol/water systems. Oppositely, greater values of D(s) were obtained for acetonitrile/water systems. It was concluded that the interaction between ODS ligands and adsorbate molecules was weaker compared with that in methanol/water mobile-phase systems when acetonitrile was used as an organic modifier in a mobile phase of reversed-phase liquid chromatography. PMID- 21639392 TI - Determination of lipophilicity by gradient elution high-performance liquid chromatography. AB - A novel method for the determination of lipophilicity using a simple HPLC protocol based on gradient elution chromatography is presented and compared to the common isocratic log k'(w) procedure. Linear relationships with high correlation coefficients between both methods for biologically active nucleosides and cyclic nucleotides as well as for environmentally relevant aromatic hydrocarbons were found. A mathematical fit to support the empirically determined linear relationship is presented. It is shown that the observed relationship between log k'(w) and the apparent capacity factor (k'(g)) determined by gradient elution is derivable by theoretical considerations as well. Since the gradient method is much less time-consuming compared to other procedures, it represents a convenient alternative for determining lipophilicity data in the future. PMID- 21639393 TI - Longitudinally modulated cryogenic system. A generally applicable approach to solute trapping and mobilization in gas chromatography. AB - This paper describes a novel approach to solute trapping and remobilization. It involves the use of a subambient trap, with a narrow capillary column passing through the cooled region. A mechanism allows for longitudinal movement of the trap relative to the chromatographic column or narrow transfer line through which analytes travel. Either the trap or the column can be moved, and the frequency or manner of relative movement determines the type of result obtained. A series of preliminary studies are described that define the general approach to using this device and demonstrate the unique benefits that may arise from the modulation movement. Significant improvement (reduction) in peak width occurs if the device is placed immediately prior to the detection system, and consequently a great increase in detection limits results. With a 25 cm length of capillary column between the trap and detector, it is calculated that the plate height of dodecane is 0.082 mm, equivalent to 12 100 plates/m. This was found to be much greater than that for dodecane analyzed on a 25 m column. By slow displacement of the column through the trap, it is demonstrated that the solute is trapped in the first 1 cm (or less) of the column located in the cold trap. PMID- 21639394 TI - Combination of column temperature gradient and mobile phase flow gradient in microcolumn and capillary column high-performance liquid chromatography. AB - The combination of several gradient modes (solvent, temperature, and flow programming) is rarely used in HPLC analysis. In this work, the separations obtained utilizing simultaneous flow and temperature gradient in capillary column and microcolumn HPLC were compared with the separations performed under isocratic, isothermal, and isorheic (constant flow) conditions. When the mobile phase flow rate and the column temperature were changed simultaneously during the separation run, the analysis time was shortened up to 50%, while the separation efficiency was preserved. The separations obtained with combined temperature and flow gradients show high reproducibility (relative standard deviation <2.0%), comparable to the reproducibility normally seen with a mobile phase gradient. For capillary HPLC, simultaneous temperature and flow programming is the method of choice because of the great technical difficulties involved in performing solvent gradient elution. PMID- 21639395 TI - Determination of detergent-derived fluorescent whitening agent isomers in lake sediments and surface waters by liquid chromatography. AB - A method for the quantitative determination of three different fluorescent whitening agents (FWAs) in lake sediments and surface waters is described. Stereoisomers of the two main laundry detergent FWAs of the diaminostilbene type (DAS 1) and of the distyrylbiphenyl type (DSBP), as well as total BLS (a compound contained in detergents until a few years ago), were quantitated in sediments and water from Greifensee, a small lake in Switzerland. The freeze-dried sediments were extracted in an ultrasonic bath using methanol with tetrabutylammonium hydrogen sulfate as an ion-pairing reagent. Aqueous samples were extracted with C18 extraction disks, which were subsequently eluted by methanol with tetrabutylammonium hydrogen sulfate. Both extracts from solid and aqueous samples were analyzed by reversed-phase high-performance liquid chromatography. Fluorescence detection was applied after postcolumn UV irradiation. Analytical reproducibility ranged from 1 to 12% (relative standard deviation). Limits of quantitation were 1-11 MUg/kg of dry matter and 0.2-3 ng/L for solid and aqueous samples, respectively. Recoveries ranged from 93 to 100% and from 87 to 95% in solid and aqueous samples, respectively. Concentrations of DAS 1 and DSBP ranged from 0.4 to 1.4 mg/kg of dry matter in top sediment layers and from 12 to 98 ng/L in lake water. Concentrations of BLS were between 0.02 and 0.08 mg/kg of dry matter in top sediment layers and <0.2 ng/L in lake water. PMID- 21639396 TI - A thermogravimetry-capillary gas chromatography/mass spectrometry interface. AB - An interface and gas chromatograph oven are described that couple a thermogravimetric analyzer with a mass spectrometer and permit multiple capillary gas chromatographic separations of volatile thermal decomposition products generated during a single thermogravimetric analysis. Examples of the use of this apparatus for identifying the volatile products generated during poly(vinyl butyral) thermal decomposition in the presence of gamma-alumina and catalytic cracking of poly(styrene) and poly(ethylene) are described. TG-GC/MS analyses employing isothermal, temperature programmed, and subambient temperature ramp gas chromatography separations are described. The apparatus permits repetitive temperature-programmed capillary gas chromatographic analyses of thermogravimetric effluent containing more than 25 constituents in 3-min intervals. PMID- 21639397 TI - Direct chromatographic separation of racemates on the basis of isotopic chirality. AB - Direct separation was achieved between two isomers which are enantiomeric to each other by virtue of the presence of hydrogen/deuterium isotopes. A racemic mixture of the R- and S-isomers of phenyl(phenyl-d(5))methanol was separated by high performance liquid chromatography using cellulose tribenzoate-coated silica as a stationary phase and a 2-propanol/hexane (5/95) mixture as a mobile phase, and the absolute configuration of each separated isomer was identified. The cellulose derivative showed preferential retention of (R)-(-)-phenyl(phenyl-d(5))methanol compared to the (S)-(+)-isomer, with a separation factor of 1.0080 based on the preferential binding of a C(6)D(5) group over a C(6)H(5) group to the primary binding site. PMID- 21639398 TI - Characterization of octadecylsilane stationary phases on commercially available silica-based packing materials by Raman spectroscopy. AB - Two commercially available solid phase extraction packing materials containing octadecylsilane stationary phases, Isolute C18MF and Isolute C18, were characterized using Raman spectroscopy. Raman spectra of excellent quality can be obtained from such systems and provide direct information about the alkyl chain conformation in the stationary phase. Data from the nu(C-C) and nu(C-H) spectral regions suggest that the alkyl chains of these stationary phases exist in a highly disordered state which is very similar to that of the neat liquid. PMID- 21639399 TI - Generating electrospray from microchip devices using electroosmotic pumping. PMID- 21639400 TI - Enhanced CO2 solubility in hybrid MCM-41: molecular simulations and experiments. AB - Grand canonical Monte Carlo simulations are performed in a hybrid adsorbent model in order to interpret the CO(2) solubility behavior. The hybrid adsorbent is prepared by confining a physical solvent (OMCTS) into the pores of a mimetic MCM 41 solid support. As a result, simulated adsorption isotherms of CO(2) nicely match the experimental data for three distinctive systems: bulk solvent, raw MCM 41, and hybrid MCM-41. The microscopic mechanisms underlying the apparition of enhanced solubility are then clearly identified. In fact, the presence of solvent molecules favors the layering of CO(2) molecules within the pores; therefore, the CO(2) solubility in the hybrid adsorbent markedly increases in comparison to that found in the raw adsorbent as well as in the bulk solvent. In addition, a good understanding of confined solvents' properties and solid surface structures is essential to fully evaluate the efficiency of hybrid adsorbents in capturing CO(2). The sorbent-solid interactions along with the solvent molecular size's impact on CO(2) solubility are therefore investigated in this study. We found that an ideal hybrid system should possess a weak solvent-solid interaction but a strong solvent-CO(2) interaction. Besides, an optimal solvent size is obtained for the enhanced CO(2) solubility in the hybrid system. According to the simulation results, the solvent layer builds pseudomicropores inside the mesoporous MCM-41, enabling more CO(2) molecules to be absorbed under the greater influence of spatial confinement and surface interaction. In addition, the molecular sieving effect is clearly observed in the case of larger solvent molecular sizes. PMID- 21639401 TI - Peculiarities of polyelectrolyte multilayer assembly on patterned surfaces. AB - The layer-by-layer assembly of poly(diallyldimethylammonium chloride) and poly(sodium 4-styrenesulfonate) is studied on templates with imprinted arrays of microwells ranging from 2 to 25 MUm and different aspect ratios. The thickness and microstructure of polyelectrolyte multilayers (PEMs) are measured using scanning electron microscopy. At 0.2 M ionic strength, the PEM film evenly coats the template both inside and outside the microwells. If the film is thinner than the critical value of about 400 nm, PEM microstructures collapse upon dissolving the template. Euler's model of critical stress is used to describe the collapse. At 2 M ionic strength, a substantially thinner PEM film is assembled inside the 25 MUm wells than outside. If the well diameter is reduced to 7 and 2 MUm, a much thicker PEM film is formed inside the microwells. These observations have been attributed to the changing of polyelectrolyte conformation in the solutions. PMID- 21639402 TI - Spatially resolved plasmonically enhanced photocurrent from Au nanoparticles on a Si nanowire. AB - Semiconducting nanowires have been demonstrated as promising light-harvesting units with enhanced absorption compared to bulk films of equivalent volume. However, for small diameter nanowires, the ultrahigh aspect ratio constrains the absorption to be polarization selective by responding primarily to the transverse magnetic (TM) light. While this effect is useful for polarization-sensitive optoelectronic devices, practical light-harvesting applications demand efficient light absorption in both TM and transverse electric (TE) light. In this study, we engineer the polarization sensitivity and the charge carrier generation in a 50 nm Si nanowire by decorating the surface with plasmonic Au nanoparticles. Using scanning photocurrent microscopy (SPCM) with a tunable wavelength laser, we spatially and spectrally resolve the local enhancement in the TE photocurrent resulting from the plasmonic near-field response of individual nanoparticles and the broad-band enhancement due to surface-enhanced absorption. These results provide guidance to the development and the optimization of nanowire-nanoparticle light-harvesting systems. PMID- 21639403 TI - Combined atomic force microscope-based topographical imaging and nanometer-scale resolved proximal probe thermal desorption/electrospray ionization-mass spectrometry. AB - Nanometer- scale proximal probe thermal desorption/electrospray ionization mass spectrometry (TD/ESI-MS) was demonstrated for molecular surface sampling of caffeine from a thin film using a 30 nm diameter nanothermal analysis (nano-TA) probe tip in an atomic force microscope (AFM) coupled via a vapor transfer line and ESI interface to a MS detection platform. Using a probe temperature of 350 degrees C and a spot sampling time of 30 s, conical desorption craters 250 nm in diameter and 100 nm deep were created as shown through subsequent topographical imaging of the surface within the same system. Automated sampling of a 5 * 2 array of spots, with 2 MUm spacing between spots, and real time selective detection of the desorbed caffeine using tandem mass spectrometry was also demonstrated. Estimated from the crater volume (~2 * 10(6) nm(3)), only about 10 amol (2 fg) of caffeine was liberated from each thermal desorption crater in the thin film. These results illustrate a relatively simple experimental setup and means to acquire in an automated fashion submicrometer scale spatial sampling resolution and mass spectral detection of materials amenable to TD. The ability to achieve MS-based chemical imaging with 250 nm scale spatial resolution with this system is anticipated. PMID- 21639404 TI - Special considerations with insulin therapy in older adults with diabetes mellitus. AB - Aging is associated with alterations in insulin secretion and action. However, aging per se does not alter the pharmacokinetics of commercially available insulin and its analogues. Insulin therapy in older adults is complicated by psychosocial and physiological changes of aging. Several new insulin and insulin analogue preparations are now available for clinical use. Used as prandial (e.g. insulin lispro, insulin aspart or insulin glulisine) and basal insulin (e.g. insulin glargine, insulin detemir), these analogues simulate physiological insulin profiles more closely than the older conventional insulins. The availability of multiple insulin products provides new opportunities to achieve control of diabetes mellitus. The choice of initial insulin therapy can be made based on blood glucose profiles. Overall, these profiles can be divided into three general patterns that include: (i) round-the-clock hyperglycaemia; (ii) fasting hyperglycaemia with daytime euglycaemia; and (iii) daytime hyperglycaemia with normal fasting blood glucose levels. The prescription of insulin is a dynamic process, and the insulin regimen should be adjusted based on individual response. The goal of diabetes care in older adults is to enhance quality of life without subjecting individuals to complicated treatment regimens that may interfere with their independence in carrying out daily activities. PMID- 21639406 TI - High concomitant use of interacting drugs and low use of gastroprotective drugs among NSAID users in an unselected elderly population: a nationwide register based study. AB - BACKGROUND: NSAIDs are commonly used in the elderly and carry significant risks of adverse events when used concomitantly with potentially interacting drugs. OBJECTIVE: The aim of this study was to provide a valid estimate of concomitant use of potentially interacting drugs and gastroprotective agents among elderly NSAID users. METHODS: This was a nationwide Swedish register-based study of drug prescriptions for 1 529 267 persons aged >=65 years in 2008, using the Swedish Prescribed Drug Register. The proportions of concomitant use of potentially interacting drugs and gastroprotective agents were analysed among NSAID users, categorized by age group, sex and NSAID use, defined as: non-users, people with 0 30 days of treatment, people with >30-180 days of treatment and people with >180 days of treatment. Use of the following concomitant drugs was evaluated: potentially harmful interacting drugs, i.e. antithrombotic drugs, antihypertensive drugs or drugs for congestive heart failure (digitalis glycosides, thiazide diuretics, loop diuretics, beta-adrenoceptor antagonists, calcium channel antagonists, ACE inhibitors and angiotensin II type 1 receptor antagonists [angiotensin receptor blockers]) and other drugs (systemic corticosteroids, selective serotonin reuptake inhibitors [SSRIs], methotrexate and lithium); and gastroprotective drugs. RESULTS: In the study cohort, 257 963 (16.9%) people had collected at least one prescription for an NSAID in 2008. This use was higher in females than in males and decreased in older age groups in both sexes. In the age groups 65-69, 70-74, 75-79, 80-84 and >=85 years, the frequencies of NSAID users were 20.6%, 20.5%, 18.9%, 15.6% and 11.6% among women, and 16.8%, 16.7%, 15.2%, 13.5% and 11.1% among men, respectively. NSAID users often concomitantly used potentially interacting drugs, e.g. over 30% used antithrombotic agents, and women had higher concomitant use of SSRIs, i.e. 12% compared with 6% in men in the youngest age group. Concomitant use of potentially harmful interacting drugs generally increased with age and with increasing use of NSAIDs. Among the eldest (age >=85 years), concomitant drug use was stable over NSAID groups. Only 40% of regular NSAID users (i.e. those prescribed NSAIDs for >180 days) were prescribed any gastroprotective medication. CONCLUSIONS: Prescribers should be alert to the possibility of potentially harmful interactions between NSAIDs and commonly prescribed drugs in the elderly. Increased use of gastroprotective medication might be justified in this context. PMID- 21639407 TI - Blood pressure-lowering efficacy of an olmesartan medoxomil/hydrochlorothiazide based treatment algorithm in elderly patients (age >=65 years) stratified by age, sex and race: subgroup analysis of a 12-week, open-label, single-arm, dose titration study. AB - INTRODUCTION: Hypertension is a leading risk factor for development of heart failure, stroke and renal disease in the elderly. OBJECTIVE: The objective of this study was to evaluate, by means of a prespecified secondary analysis of a 12 week, open-label, single-arm, dose-titration study, the blood pressure (BP) lowering efficacy and safety of an olmesartan medoxomil (OM)/hydrochlorothiazide (HCTZ)-based titration regimen in patients aged >=65 years with hypertension. Subgroups were stratified by age (>=65 to <=75 or >75 years), sex (male or female) and race (Black or non-Black). METHODS: Following a 2- to 3-week placebo run-in phase, patients received OM 20 mg, uptitrated to OM 40 mg, followed by addition of HCTZ 12.5-25 mg step-wise at 3-week intervals if seated cuff BP (SeBP) was >=120/70 mmHg. Patients below this target SeBP were maintained at their current dose but uptitrated to the next consecutive dose if mean seated cuff systolic BP (SBP) was >=140 mmHg and/or mean seated cuff diastolic BP was >=90 mmHg at follow-up visits. Efficacy was assessed by 24-hour ambulatory BP monitoring (ABPM) and SeBP measurements. The primary efficacy variable was the change from baseline in mean 24-hour ambulatory SBP after 12 weeks. Secondary efficacy endpoints included the change from baseline in mean 24-hour ambulatory SBP; change from baseline in ambulatory BP during the daytime (8:00 am-4:00 pm), nighttime (10:00 pm-6:00 am) and the last 6, 4 and 2 hours of the dosing interval; change from baseline in SeBP at each titration step and at study end; and the proportion of patients achieving mean 24-hour ambulatory BP targets and SeBP goals at week 12. The frequency and severity of treatment-emergent adverse events (TEAEs) were also documented. RESULTS: Baseline and week 12 ABPM data were available for 150 out of 178patients who entered the active treatment phase. Changes from baseline in mean 24-hour ambulatory BP were -26.0/-12.5 mmHg and 24.9/-12.0 mmHg in patients aged >=65 to <=75 years (n = 128) and >75 years (n = 48), respectively (all p < 0.0001 vs baseline). Changes from baseline in mean 24 hour ambulatory BP were -26.0/-13.0 mmHg and -25.4/-11.5 mmHg in male (n = 92) and female (n = 84) patients, respectively (all p < 0.0001 vs baseline) and 26.7/-11.8 mmHg and -25.6/-12.4 mmHg in Black (n = 28) and non-Black (n = 148) patients, respectively (all p < 0.0001 vs baseline). Clinically significant ambulatory BP reductions were observed during the daytime, nighttime and the last 6, 4 and 2 hours of the dosing interval in all subgroups. Changes from baseline at week 12 in mean SeBP were similar to 24-hour ambulatory BP changes reported previously. At week 12, the proportion of patients achieving the 24-hour ambulatory BP target of <130/80 mmHg ranged from 67.5% to 77.4% and achieving the SeBP goal of <140/90 mmHg ranged from 60.7% to 68.8% across the subgroups. Most TEAEs and drug-related TEAEs were mild or moderate in severity, and there were no trends across subgroups. CONCLUSIONS: In a subgroup analysis based upon age, sex and race in patients aged >=65 years with hypertension, an OM/HCTZ-based algorithm was efficacious and well tolerated. ClinicalTrials.gov Identifier: NCT00412932. PMID- 21639405 TI - The role of phytochemicals in the treatment and prevention of dementia. AB - Dementia pathologies such as Alzheimer's disease (AD) are reaching epidemic proportions, yet they are not successfully managed by effective symptomatic treatments. Only five drugs have been developed to alleviate cognitive symptoms, and more effective and safe treatments are needed for both the cognitive symptoms and behavioural and psychological symptoms of dementia (BPSD). As two of these licensed drugs (cholinesterase inhibitors [ChEIs]) are naturally derived (galantamine and rivastigmine), the potential for plants to yield new therapeutic agents has stimulated extensive research to discover new ChEIs together with plant extracts, phytochemicals and their derivatives with other mechanistic effects relevant to dementia treatment. This review presents the potential and actual therapeutic strategies for dementia in relation to the known mechanisms of dementia pathology. Phytochemicals that have shown mechanistic effects relevant to the pathological targets in dementia are discussed, with an emphasis on those showing positive clinical trial evidence. Those phytochemicals discussed include the alkaloid physostigmine, a ChEI from the calabar bean (Physostigma venenosum), which has been used as a template for the development of synthetic derivatives that inhibit acetylcholinesterase, including the drug rivastigmine. Also discussed are other ChEI alkaloids including huperzine A, from Huperzia serrata, and galantamine, originally from the snowdrop (Galanthus woronowii); both alkaloids improve cognitive functions in AD patients. Other phytochemicals discussed include cannabinoids (e.g. cannabidiol) from Cannabis sativa, which are emerging as potential therapeutic agents for BPSD, and resveratrol (occurs in various plants) and curcumin (from turmeric [Curcuma longa]), which have been investigated for their pharmacological activities relevant to dementia and their potential effects on delaying dementia progression. The review also discusses plant extracts, and their known constituents, that have shown relevant mechanistic effects for dementia and promising clinical data, but require more evidence for their clinical efficacy and safety. Such plants include Ginkgo biloba, which has been extensively studied in numerous clinical trials, with most outcomes showing positive effects on cognitive functions in dementia patients; however, more reliable and consistent clinical data are needed to confirm efficacy. Other plants and their extracts that have produced promising clinical data in dementia patients, with respect to cognition, include saffron (Crocus sativus), ginseng (Panax species), sage (Salvia species) and lemon balm (Melissa officinalis), although more extensive and reliable clinical data are required. Other plants that are used in traditional practices of medicine have been suggested to improve cognitive functions (e.g. Polygala tenuifolia) or have been associated with alleviation of BPSD (e.g. the traditional prescription yokukansan); such remedies are often prescribed as complex mixtures of different plants, which complicates interpretation of pharmacological and clinical data and introduces additional challenges for quality control. Evidence for the role of natural products in disease prevention, the primary but considerably challenging aim with respect to dementia, is limited, but the available epidemiological and clinical evidence is discussed, with most studies focused on ChEIs, nicotine (from Nicotiana species), curcumin, wine polyphenols such as resveratrol and G. biloba. Challenges for the development of phytochemicals as drugs and for quality control of standardized plant extracts are also considered. PMID- 21639408 TI - Risk factors for prescribing and transcribing medication errors among elderly patients during acute hospitalization: a cohort, case-control study. AB - BACKGROUND: Medication errors are the cause of common and dangerous adverse effects on patients. They occur as a result of preventable failures in the prescribing (ordering), transcribing, dispensing and administration of medications. OBJECTIVE: The aim of this study was to determine the relationship between patient characteristics and prescribing and transcribing medication errors during acute hospitalization of elderly patients in an internal medicine ward. METHODS: A cohort case-control study was conducted in a 37-bed medical ward at a tertiary medical centre in Israel. The study included 137 patients in whom potentially harmful medication errors were detected, and 137 sex- and age-matched controls. Clinical data were collected and Charlson Comorbidity Index scores were calculated. Conditional logistic regression was used to identify factors associated with medication errors. RESULTS: Independent factors associated with any type of medication error included Charlson Comorbidity Index score >=8 (odds ratio [OR] 2.97 [95% CI 1.16, 7.61], p = 0.023), number of medications >= 9 (OR 2.02 [95% CI 1.00, 4.05], p = 0.049) [both upon admission] and length of hospital stay >= 13 days (OR 4.41 [95% CI 2.25, 8.62], p < 0.0001). Independent factors associated with prescribing errors included Charlson Comorbidity Index score >= 8 (OR 6.34 [95% CI 1.63, 24.71], p = 0.008) and length of hospital stay >=13 days (OR 3.19 [95% CI 1.23, 8.26], p = 0.017), while independent factors associated with transcribing errors included number of medications >=9 (OR 2.58 [95% CI 1.02, 6.51], p = 0.04) and length of hospital stay >= 13 days (OR 6.90 [95% CI 2.76, 17.23], p < 0.0001). The median time to an error was 3 days, and was half as long for prescribing errors as for transcribing errors (2 and 4 days, respectively, p = 0.017). CONCLUSIONS: The risk of medication errors among elderly patients during acute hospitalization in an internal medicine ward is associated with Charlson Comorbidity Index score (for prescribing errors), number of medications (for transcribing errors) and length of hospital stay (for both types of errors). Further study will determine whether these factors can be used to identify patients at risk and to prevent prescribing and transcribing medication errors. PMID- 21639409 TI - Spotlight on intravenous vernakalant in recent-onset atrial fibrillation. AB - Intravenous vernakalant (Brinavess(r)) is an atrial-repolarization-delaying agent that is currently approved in the EU for the rapid conversion of recent-onset atrial fibrillation to sinus rhythm. Vernakalant blocks atrial-specific potassium and sodium ion channels, prolonging atrial refractory periods and rate dependently slowing atrial conduction, without promoting ventricular arrhythmia. In pivotal, randomized, phase III trials, intravenous vernakalant 3 mg /kg administered as a 10-minute infusion, followed by a 2 mg/kg 10-minute infusion after 15 minutes if atrial fibrillation persisted, was effective in the rapid termination of recent-onset atrial fibrillation in nonsurgical patients (>= 3 hours' to <= 7 days' duration) and in those with postoperative atrial fibrillation (3-72 hours' duration) following cardiac surgery. Conversion to sinus rhythm occurred rapidly following infusion of vernakalant, with the majority of patients converting after the first dose, and conversion to sinus rhythm was generally associated with a rapid resolution of symptoms. These antiarrhythmic effects of vernakalant were durable, with most responders remaining in sinus rhythm 24 hours after treatment initiation. In nonsurgical patients with recent-onset atrial fibrillation of 3-48 hours' duration, vernakalant was more effective than intravenous amiodarone, with a significantly higher proportion of patients converting to sinus rhythm within the first 90 minutes of treatment. Vernakalant was generally well tolerated in clinical trials, with most adverse events being of mild or moderate severity and not treatment limiting. Increases in QRS or QT intervals were transient, and there was no increased incidence of ventricular arrhythmia observed with vernakalant compared with placebo. Therefore, intravenous vernakalant provides an effective option for the management of recent-onset atrial fibrillation. PMID- 21639411 TI - Inactivated influenza vaccines: pre-vaccination haemagglutinin-antibody titres influence the vaccine response but not necessarily the vaccine effectiveness. PMID- 21639412 TI - Communication: atomic and molecular Rydbergs from water. AB - We report the formation of energetic neutral Rydberg hydrogen atoms and transient Rydberg molecular ions, [(H(2)O)(q+)](?) in ion-impact dissociation of isolated water molecules. The kinetic energy spectra of the neutral Rydberg H atoms are determined from the complete study of (H(?), H(+), O(+)) dissociation channel. This channel of water dissociation is suggested as a possible additional source of the energetic neutrals detected in upper atmospheres of extra solar planets, and of slow electrons which are known to play a major role in radiation induced damage to living cells. PMID- 21639413 TI - Communication: spectroscopic measurements for HfF+ of relevance to the investigation of fundamental constants. AB - The properties of the HfF(+) cation are thought to be well-suited for investigations of the electron electric dipole moment (eEDM) and temporal variations of the fine structure constant. Precision spectroscopic measurements involving the X(1)Sigma(+) and low-lying (3)Delta(1) states have been proposed to measure both. Due to the lack of data for HfF(+), the design of these experiments has relied entirely on the predictions of electronic structure calculations. Spectroscopic characterizations of the X(1)Sigma(+), (3)Delta(1), (3)Delta(2) and (3)Delta(3) states are reported. The results further support the contention that HfF(+) is a viable candidate for eEDM measurements. The spacings between adjacent X(1)Sigma(+) and (3)Delta(1) levels are found to be less favorable for the proposed studies of the fine structure constant. PMID- 21639414 TI - Communication: equation of state of hard oblate ellipsoids by replica exchange Monte Carlo. AB - We implemented the replica exchange Monte Carlo technique to produce the equation of state of hard 1:5 aspect-ratio oblate ellipsoids for a wide density range. For this purpose, we considered the analytical approximation of the overlap distance given by Bern and Pechukas and the exact numerical solution given by Perram and Wertheim. For both cases we capture the expected isotropic-nematic transition at low densities and a nematic-crystal transition at larger densities. For the exact case, these transitions occur at the volume fraction 0.341, and in the interval 0.584-0.605, respectively. PMID- 21639415 TI - Communication: mode-specific photodissociation of vibrationally excited pyrrole. AB - Laser-based spectroscopies coupled with molecular beam techniques facilitated the monitoring of H fragments released in ultraviolet photodissociation of pre excited isoenergetic vibrational levels of pyrrole. Most noticeably, there was an order of magnitude larger reactivity for an eigenstate primarily consisting of two quanta of ring deformation than for another with one quantum of symmetric C-H stretch. The dynamics, the intramolecular interactions controlling the energy flow, and the mode-selectivity within a medium-sized, ten atom molecule, is discussed. PMID- 21639416 TI - Communication: rovibrationally selected study of the N2+(X; v+=1, N+= 0-8) + Ar charge transfer reaction using the vacuum ultraviolet laser pulsed field ionization-photoion method. AB - By employing an electric field pulsing scheme for vacuum ultraviolet laser pulsed field ionization-photoion (PFI-PI) measurements, we have been able to prepare a rovibrationally selected PFI-PI beam of N(2)(+)(v(+) = 1, N(+)) with not only high intensity and high quantum state purity, but also high kinetic energy resolution, allowing absolute total cross sections [sigma(v(+) = 1, N(+))] for the N(2)(+)(X; v(+) = 1, N(+)) + Ar, N(+) = 0-8 charge transfer reaction to be measured at center-of-mass collision energies (E(cm)) down to thermal energies. The sigma(v(+) = 1, N(+) = 0-8) values determined at E(cm) = 0.04-10.00 eV are in good agreement with the theoretical predictions based on the Landau-Zener Stuckelberg formulism. Taking into account the experimental uncertainties, the sigma(v(+) = 1, N(+)), N(+) = 0-8, measured at E(cm) = 1.56 eV are found to be independent of N(+). PMID- 21639417 TI - Communication: electric dipole moment and hyperfine interaction of tungsten monocarbide, WC. AB - The [17.6]2<- X(3)Delta(1)(1,0) band of tungsten monocarbide, WC, was recorded using laser induced fluorescence both field-free and in the presence of a static electric field. The Stark induced shifts of the R(1) and Q(2) lines were analyzed to produce values for the permanent electric dipole moments, MU, of 3.90 +/- 0.04 D and 2.57 +/- 0.04 D for the X(3)Delta(1)(v = 0) and [17.6]2(v = 1) states, respectively. An upper limit to the Omega-doubling parameter, o(Delta), of 2 MHz is established. The (183)W(I = 1/2) Fermi contact parameter, b(F), for the X(3)Delta(1)(v = 0) is estimated to be 1363 MHz. The results are compared with theoretical predictions and a molecular orbital correlation description. PMID- 21639418 TI - A study of complex scaling transformation using the Wigner representation of wavefunctions. AB - The complex scaling operator exp(-theta xp/h), being a foundation of the complex scaling method for resonances, is studied in the Wigner phase-space representation. It is shown that the complex scaling operator behaves similarly to the squeezing operator, rotating and amplifying Wigner quasi-probability distributions of the respective wavefunctions. It is disclosed that the distorting effect of the complex scaling transformation is correlated with increased numerical errors of computed resonance energies and widths. The behavior of the numerical error is demonstrated for a computation of CO(2+) vibronic resonances. PMID- 21639419 TI - Breakdown of the pseudopotential approximation for magnetizabilities and electric multipole moments: test calculations for Au, AuF, and Sn(n) cluster (n <= 20). AB - The response of the electronic wavefunction to an external electric or magnetic field is widely considered to be a typical valence property and should, therefore, be adequately described by accurately adjusted pseudopotentials, especially if a small-core definition is used within this approximation. In this paper we show for atomic Au and Au(+), as well as for the molecule AuF and tin clusters, that in contrast to the case of the static electric dipole polarizability or the electric dipole moment, core contributions to the static magnetizability are non-negligible, and can therefore lead to erroneous results within the pseudopotential approximation. This error increases with increasing size of the core chosen. For tin clusters, which are of interest in ongoing molecular beam experiments currently carried out by the Darmstadt group, the diamagnetic and paramagnetic isotropic components of the magnetizability tensor almost cancel out and large-core pseudopotentials do not even predict the correct sign for this property due to erroneous results in both the diamagnetic and (more importantly) the paramagnetic terms. Hence, all-electron calculations or pseudopotentials with very small cores are required to adequately predict magnetizabilities for atoms, molecules and the solid state, making it computationally more difficult to obtain this quantity for future investigations in heavy atom containing molecules or clusters. We also demonstrate for this property that all-electron density functional calculations are quite robust and give results close to wavefunction based methods for the atoms and molecules studied here. PMID- 21639420 TI - Integrating steepest-descent reaction pathways for large molecules. AB - Exploring potential energy surfaces of large molecular systems can be quite challenging due to the increased number of nuclear degrees of freedom. Many techniques that are well-suited for small and moderate size systems require diagonalization of the energy second-derivative matrix. Since the cost of this step scales as O(N(atoms)(3)) (where N(atoms) is the number of atomic centers), such methods quickly become infeasible and are eventually rendered cost prohibitive. In this work, the recently developed Euler-based predictor-corrector reaction path integration method [H. P. Hratchian, M. J. Frisch, and H. B. Schlegel, J. Chem. Phys. 133, 224101 (2010)] is enhanced and proposed as a useful alternative to conventional reaction path following schemes in studies on very large systems. Because this integrator does not require Hessian diagonalization, the O(N(atoms)(3)) bottleneck afflicting other approaches is completely avoided. The effectiveness of the integrator in large system studies is demonstrated with an enzyme-catalyzed reaction employing an ONIOM (QM:MM) model chemistry and involving 5368 atomic centers. PMID- 21639421 TI - Adiabatic versus diabatic descriptions of the lowest Rydberg and valence 1Sigma+ states of HCl. AB - In this contribution we first report new ab initio self-consistent field configuration interaction calculations of the first excited adiabatic potential of (1)Sigma(+) symmetry, the 2(1)Sigma(+) or B(1)Sigma(+) state, which presents two minima and can thus be seen as made up of the Rydberg E(1)Sigma(+) and the valence V(1)Sigma(+) states. Based on the computed 2(1)Sigma(+) potential, we devised a theoretical procedure to compute the vibronic structure in order to try to explain the energy levels observed in the region above 76 254.4 cm(-1) which display an irregular vibrational structure, indicative of spectral perturbations. We try to find out which representation of the electronic states, the diabatic or the adiabatic one, is best suited to replicate the lowest observed vibronic levels of the E and V states. To this end, we deduce, from the 2(1)Sigma(+) potential and its complementary adiabatic potential, two diabatic potentials. We then carry out a coupled equation treatment based on these diabatic potentials. The results of this treatment indicate that, in the present case, the adiabatic representation is better than the diabatic one to describe the observed vibronic levels. This is due, as expected, to the existence of a strong electrostatic interaction between the two diabatic potentials. PMID- 21639422 TI - Markov state models based on milestoning. AB - Markov state models (MSMs) have become the tool of choice to analyze large amounts of molecular dynamics data by approximating them as a Markov jump process between suitably predefined states. Here we investigate "Core Set MSMs," a new type of MSMs that build on metastable core sets acting as milestones for tracing the rare event kinetics. We present a thorough analysis of Core Set MSMs based on the existing milestoning framework, Bayesian estimation methods and Transition Path Theory (TPT). We show that Core Set MSMs can be used to extract phenomenological rate constants between the metastable sets of the system and to approximate the evolution of certain key observables. The performance of Core Set MSMs in comparison to standard MSMs is analyzed and illustrated on a toy example and in the context of the torsion angle dynamics of alanine dipeptide. PMID- 21639423 TI - Fourth-order relativistic corrections to electrical first-order properties using direct perturbation theory. AB - In this work, we present relativistic corrections to first-order electrical properties obtained using fourth-order direct perturbation theory (DPT4) at the Hartree-Fock level. The considered properties, i.e., dipole moments and electrical-field gradients, have been calculated using numerical differentiation techniques based on a recently reported DPT4 code for energies [S. Stopkowicz and J. Gauss, J. Chem. Phys. 134, 064114 (2011)]. For the hydrogen halides HX, X=F, Cl, Br, I, and At, we study the convergence of the scalar-relativistic contributions by comparing the computed DPT corrections to results from spin-free Dirac-Hartree-Fock calculations. Furthermore, since in the DPT series spin-orbit contributions first appear at fourth order, we investigate their magnitude and judge the performance of the DPT4 treatment by means of Dirac-Hartree-Fock benchmark calculations. Finally, motivated by experimental investigations of the molecules CH(2)FBr, CHF(2)Br, and CH(2)FI, we present theoretical results for their halogen quadrupole-coupling tensors and give recommendations concerning the importance of higher-order scalar-relativistic and spin-orbit corrections. PMID- 21639424 TI - Third-order corrections to random-phase approximation correlation energies. AB - Several random-phase approximation (RPA) correlation methods were compared in third order of perturbation theory. While all of the considered approaches are exact in second order of perturbation theory, it is found that their corresponding third-order correlation energy contributions strongly differ from the exact third-order correlation energy contribution due to missing interactions of the particle-particle-hole-hole type. Thus a simple correction method is derived which makes the different RPA methods also exact to third-order of perturbation theory. By studying the reaction energies of 16 chemical reactions for 21 small organic molecules and intermolecular interaction energies of 23 intermolecular complexes comprising weakly bound and hydrogen-bridged systems, it is found that the third-order correlation energy correction considerably improves the accuracy of RPA methods if compared to coupled-cluster singles doubles with perturbative triples as a reference. PMID- 21639425 TI - Configuration selection within vibrational multiconfiguration self-consistent field theory: application to bridged lithium compounds. AB - A configuration selection scheme has been used to speed up vibrational multiconfiguration self-consistent field calculations. Deviations with respect to reference calculations were found to be negligible while yielding an acceleration of about two orders of magnitude. Its application to bridged lithium compounds (Li(2)H(2), Li(2)F(2), Li(2)O(2), and Li(3)F(3)) based on high-level coupled cluster potential energy surfaces provides accurate vibrational transitions for all fundamental modes. The explicit inclusion of 4-mode couplings was found to be important for Li(2)H(2). PMID- 21639426 TI - Electronic coupling calculation and pathway analysis of electron transfer reaction using ab initio fragment-based method. I. FMO-LCMO approach. AB - By making use of an ab initio fragment-based electronic structure method, fragment molecular orbital-linear combination of MOs of the fragments (FMO-LCMO), developed by Tsuneyuki et al. [Chem. Phys. Lett. 476, 104 (2009)], we propose a novel approach to describe long-distance electron transfer (ET) in large system. The FMO-LCMO method produces one-electron Hamiltonian of whole system using the output of the FMO calculation with computational cost much lower than conventional all-electron calculations. Diagonalizing the FMO-LCMO Hamiltonian matrix, the molecular orbitals (MOs) of the whole system can be described by the LCMOs. In our approach, electronic coupling T(DA) of ET is calculated from the energy splitting of the frontier MOs of whole system or perturbation method in terms of the FMO-LCMO Hamiltonian matrix. Moreover, taking into account only the valence MOs of the fragments, we can considerably reduce computational cost to evaluate T(DA). Our approach was tested on four different kinds of model ET systems with non-covalent stacks of methane, non-covalent stacks of benzene, trans-alkanes, and alanine polypeptides as their bridge molecules, respectively. As a result, it reproduced reasonable T(DA) for all cases compared to the reference all-electron calculations. Furthermore, the tunneling pathway at fragment-based resolution was obtained from the tunneling current method with the FMO-LCMO Hamiltonian matrix. PMID- 21639427 TI - A simple polarizable continuum solvation model for electrolyte solutions. AB - We propose a Debye-Huckel-like screening model (DESMO) that generalizes the familiar conductor-like screening model (COSMO) to solvents with non-zero ionic strength and furthermore provides a numerical generalization of the Debye-Huckel model that is applicable to non-spherical solute cavities. The numerical implementation of DESMO is based upon the switching/Gaussian (SWIG) method for smooth cavity discretization, which we have recently introduced in the context of polarizable continuum models (PCMs). This approach guarantees that the potential energy is a smooth function of the solute geometry and analytic gradients for DESMO are reported here. The SWIG formalism also facilitates analytic implementation of two other PCMs that are based on a screened Coulomb potential: the "integral equation formalism" (IEF-PCM) and the "surface and simulation of volume polarization for electrostatics" [SS(V)PE] method. Fully analytic implementations of these screened PCMs are reported here for the first time. Numerical results, for model systems where an exact solution of the linearized Poisson-Boltzmann equation is available, demonstrate that these screened PCMs are highly accurate. In realistic test cases, they are as accurate as the best available three-dimensional finite-difference methods. In polar solvents, DESMO is nearly as accurate as more sophisticated screened PCMs, but is significantly simpler and more efficient. PMID- 21639428 TI - Pilot applications of internally contracted multireference coupled cluster theory, and how to choose the cluster operator properly. AB - The internally contracted multireference coupled cluster (icMRCC) method allows a highly accurate description of both static and dynamic correlation with a computational scaling similar to single reference coupled cluster theory. The authors show that the method can lose its orbital invariance and size consistency when no special care is taken in the elimination of redundant excitations. Using the BeH(2) model system, four schemes are compared which differ in their treatment of linear dependencies between excitations of different rank (such as between singles and doubles). While the energy curves agree within tens of MUE(h) when truncating the cluster operator at double excitations (icMRCCSD), inclusion of triple excitations (icMRCCSDT) leads to significant differences of more than 1 mE(h). One scheme clearly yields the best results, while the others even turn out to be not size consistent. The former procedure uses genuine single and double excitations and discards those linear combinations of (spectator) double and triple excitations which have the same effect on the reference function. With this approach, the equilibrium structure and harmonic vibrational frequencies of ozone obtained with icMRCCSDT are in excellent agreement with CCSDTQ. The authors further apply icMRCC methods to potential energy surfaces of HF, LiF, N(2), and to the singlet-triplet splitting of benzynes. In particular, the latter calculations have been made possible by implementing the method with the proper formal scaling using automated techniques. PMID- 21639429 TI - Local control of non-adiabatic dissociation dynamics. AB - We present a theoretical approach which consists of applying the strategy of local control to projectors based on asymptotic scattering states. This allows to optimize final state distributions upon laser excitation in cases where strong non-adiabatic effects are present. The approach, despite being based on a time local formulation, can take non-adiabatic transitions that appear at later times fully into account and adopt a corresponding control strategy. As an example, we show various dissociation channels of HeH(+), a system where the ultrafast dissociation dynamics is determined by strong non-Born-Oppenheimer effects. PMID- 21639430 TI - A proof of Clausius' theorem for time reversible deterministic microscopic dynamics. AB - In 1854 Clausius proved the famous theorem that bears his name by assuming the second "law" of thermodynamics. In the present paper we give a proof that requires no such assumption. Our proof rests on the laws of mechanics, a T-mixing property, an ergodic consistency condition, and on the axiom of causality. Our result relies on some recently derived theorems, such as the Evans-Searles and the Crooks fluctuation theorems and the recently discovered relaxation and dissipation theorems. PMID- 21639431 TI - Many-body dissipative particle dynamics simulation of liquid/vapor and liquid/solid interactions. AB - The combination of short-range repulsive and long-range attractive forces in many body dissipative particle dynamics (MDPD) is examined at a vapor/liquid and liquid/solid interface. Based on the radial distribution of the virial pressure in a drop at equilibrium, a systematic study is carried out to characterize the sensitivity of the surface tension coefficient with respect to the inter-particle interaction parameters. For the first time, the approximately cubic dependence of the surface tension coefficient on the bulk density of the fluid is evidenced. In capillary flow, MDPD solutions are shown to satisfy the condition on the wavelength of an axial disturbance leading to the pinch-off of a cylindrical liquid thread; correctly, no pinch-off occurs below the cutoff wavelength. Moreover, in an example that illustrates the cascade of fluid dynamics behaviors from potential to inertial-viscous to stochastic flow, the dynamics of the jet radius is consistent with the power law predictions of asymptotic analysis. To model interaction with a solid wall, MDPD is augmented by a set of bell-shaped weight functions; hydrophilic and hydrophobic behaviors, including the occurrence of slip in the latter, are reproduced using a modification in the weight function that avoids particle clustering. The dynamics of droplets entering an inverted Y shaped fracture junction is shown to be correctly captured in simulations parametrized by the Bond number, confirming the flexibility of MDPD in modeling interface-dominated flows. PMID- 21639432 TI - Conical intersections in solution: formulation, algorithm, and implementation with combined quantum mechanics/molecular mechanics method. AB - The significance of conical intersections in photophysics, photochemistry, and photodissociation of polyatomic molecules in gas phase has been demonstrated by numerous experimental and theoretical studies. Optimization of conical intersections of small- and medium-size molecules in gas phase has currently become a routine optimization process, as it has been implemented in many electronic structure packages. However, optimization of conical intersections of small- and medium-size molecules in solution or macromolecules remains inefficient, even poorly defined, due to large number of degrees of freedom and costly evaluations of gradient difference and nonadiabatic coupling vectors. In this work, based on the sequential quantum mechanics and molecular mechanics (QM/MM) and QM/MM-minimum free energy path methods, we have designed two conical intersection optimization methods for small- and medium-size molecules in solution or macromolecules. The first one is sequential QM conical intersection optimization and MM minimization for potential energy surfaces; the second one is sequential QM conical intersection optimization and MM sampling for potential of mean force surfaces, i.e., free energy surfaces. In such methods, the region where electronic structures change remarkably is placed into the QM subsystem, while the rest of the system is placed into the MM subsystem; thus, dimensionalities of gradient difference and nonadiabatic coupling vectors are decreased due to the relatively small QM subsystem. Furthermore, in comparison with the concurrent optimization scheme, sequential QM conical intersection optimization and MM minimization or sampling reduce the number of evaluations of gradient difference and nonadiabatic coupling vectors because these vectors need to be calculated only when the QM subsystem moves, independent of the MM minimization or sampling. Taken together, costly evaluations of gradient difference and nonadiabatic coupling vectors in solution or macromolecules can be reduced significantly. Test optimizations of conical intersections of cyclopropanone and acetaldehyde in aqueous solution have been carried out successfully. PMID- 21639433 TI - Effective surface motion on a reactive cylinder of particles that perform intermittent bulk diffusion. AB - In many biological and small scale technological applications particles may transiently bind to a cylindrical surface. In between two binding events the particles diffuse in the bulk, thus producing an effective translation on the cylindrical surface. We here derive the effective motion on the surface allowing for additional diffusion on the cylindrical surface itself. We find explicit solutions for the number of adsorbed particles at one given instant, the effective surface displacement, as well as the surface propagator. In particular sub- and superdiffusive regimes are found, as well as an effective stalling of diffusion visible as a plateau in the mean squared displacement. We also investigate the corresponding first passage problem. PMID- 21639434 TI - Dissociative excitation and fragmentation of S8 by electron impact. AB - The vacuum-ultraviolet emission spectrum from 136 nm to 168 nm following the dissociative excitation of a predominantly S(8) target by electron impact at 100 eV incident energy was measured. The relative cross sections for the dominant multiplets at 138.9, 142.9, 147.9, and 166.7 nm are presented. Excitation functions are shown for electron-impact energies from below threshold to 360 eV for the two most prominent emissions at 142.5 nm and 147.4 nm. Five thresholds are clearly apparent in both excitation functions. For the four highest energy channels, the energy separation between the adjacent thresholds is approximately constant and the cross sections reduce regularly as the threshold energies increase. We suggest possible fragmentation pathways of the dissociating S(8) molecule that reproduce the energies of our observed thresholds. PMID- 21639435 TI - Photodissociation investigation of doubly charged ethanol clusters induced by inner-shell electron ionization. AB - Fragmentation of doubly charged ethanol clusters [(C(2)H(5)OH)(n)] following the O 1s ionization has been investigated by means of the photoelectron-photoion photoion coincidence (PEPIPICO) method. The dominant fission channel of (C(2)H(5)OH)(n)(2+) was the formation of protonated cluster ion pairs [H(C(2)H(5)OH)(l)(+)/H(C(2)H(5)OH)(m)(+)]. The fragmentation mechanisms of these ion pairs were discussed based on the analysis of the PEPIPICO contour shape. It was clarified that the prominent fragmentation channel was a secondary decay mechanism, where neutral evaporation occurs after charge separation. On the other hand, the formation of small fragment ions was suppressed, excluding the formation of certain specific fragments (H(3)O(+), C(2)H(5)(+)/COH(+), and C(2)H(4)OH(+)). The formation of small fragment ions was suppressed due to the cooling effect caused by the neutral evaporation and the decrease in the electrostatic repulsive force caused by charge separation. PMID- 21639436 TI - Photoelectron imaging spectroscopy and theoretical investigation of ZrSi. AB - The photoelectron spectrum of ZrSi(-) has been measured at two different photon energies: 2.33 eV and 3.49 eV, providing electron binding energy and photoelectron angular distribution information. The obtained vertical detachment energy of ZrSi(-) is 1.584(14) eV. The neutral ground and excited state terms are assigned based on experimental and theoretical results. The ground state of ZrSi is tentatively assigned as a (3)Sigma(+) state with a configuration of 1sigma(2) 1pi(4) 1delta(0) 2sigma(1) 3sigma(1). A low lying (3)Pi(i) neutral excited state is identified to be 0.238 eV (1919 cm(-1)) above the ground state. The anion ground state is designated as a (2)Sigma(+) state with a 1sigma(2) 1pi(4) 1delta(0) 2sigma(2) 3sigma(1) valence electron configuration. A Franck-Condon (FC) simulation of the photoelectron spectrum has been carried out. For the (3)Sigma(+) <- (2)Sigma(+) band, theoretically calculated bond lengths and frequencies are used in the FC calculation which give good agreement with experiment, while for the (3)Pi(i) <- (2)Sigma(+) band, the ZrSi bond length is estimated from the FC spectrum. Comparisons are made with previously published theoretical studies and inconsistencies are pointed out. To the best of our knowledge, this study provides the first spectroscopic information on the transition metal-silicon diatomic, ZrSi. PMID- 21639437 TI - Dyson orbitals of N2O: electron momentum spectroscopy and symmetry adapted cluster-configuration interaction calculations. AB - Electron momentum spectroscopy and symmetry adapted cluster-configuration interaction (SAC-CI) theory were combined to study electron correlation effects in nitrous oxide molecule (N(2)O). The SAC-CI General-R method accurately reproduced the experimental ionization spectrum. This bench-marked method was also introduced for calculating the momentum distributions of N(2)O Dyson orbitals. Several calculated momentum distributions with different theoretical methods were compared with the high resolution experimental results. In the outer valence region, Hartree-Fock (HF), density functional theory (DFT), and SAC-CI theory can well describe the experimental momentum distributions. SAC-CI presented a best performance among them. In the inner-valence region, HF and DFT cannot work well due to the severe breaking of the molecular orbital picture, while SAC-CI still produced an excellent description of experimental momentum profiles because it can accurately take into account electron correlations. Moreover, the thermally averaged calculation showed that the geometrical changes induced by the vibration at room temperature have no noticeable effects on momentum distribution of valence orbitals of N(2)O. PMID- 21639438 TI - Theory of vibronic interactions in D2 and H2: a comparison between multichannel quantum-defect and coupled-equation approaches. AB - New experimental energy levels for the 2ppiC(1)Pi(u)(-) state of D(2) are reported extending up to the dissociation limit and including rotational quantum numbers up to N = 10. These data are extracted from recent high resolution optical emission spectra, and they are used for a detailed comparison of two theoretical approaches, both of which are fully ab initio and are based on the same state-of-the-art clamped-nuclei potential energy curves. These are the coupled differential equations (CE) and the multichannel quantum defect theory (MQDT) approaches, each of which accounts for adiabatic corrections and non adiabatic couplings. Both theoretical approaches reproduce the experimental levels to within a fraction of a wavenumber unit (cm(-1)) for the lower vibrational quantum numbers, with the MQDT surpassing the CE method. As the dissociation limit is approached, the residuals observed-calculated increase up to several cm(-1) and the MQDT method is up to a factor of two less accurate than the CE method. The same analysis is carried out with existing data for the H(2) isotopomer and yields similar results. An analogous comparison is also made for the 3ppiD(1)Pi(u)(-) and 4ppiD('1)Pi(u)(-) states for both isotopomers, where the MQDT is found to be superior to the CE approach. PMID- 21639439 TI - Electronic states of thiophenyl and furanyl radicals and dissociation energy of thiophene via photoelectron imaging of negative ions. AB - We report photoelectron images and spectra of deprotonated thiophene, C(4)H(3)S( ), obtained at 266, 355, and 390 nm. Photodetachment of the alpha isomer of the anion is observed, and the photoelectron bands are assigned to the ground X(2)A(') (sigma) and excited A(2)A(") and B(2)A(") (pi) states of the thiophenyl radical. The photoelectron angular distributions are consistent with photodetachment from the respective in-plane (sigma) and out-of-plane (pi(*)) orbitals. The adiabatic electron affinity of alpha-(?)C(4)H(3)S is determined to be 2.05 +/- 0.08 eV, while the B(2)A(") term energy is estimated at 1.6 +/- 0.1 eV. Using the measured electron affinity and the electron affinity/acidity thermodynamic cycle, the C-H(alpha) bond dissociation energy of thiophene is calculated as DH(298)(H(alpha)-C(4)H(3)S) = 115 +/- 3 kcal/mol. Comparison of this value to other, previously reported C-H bond dissociation energies, in particular for benzene and furan, sheds light of the relative thermodynamic stabilities of the corresponding radicals. In addition, the 266 nm photoelectron image and spectrum of the furanide anion, C(4)H(3)O(-), reveal a previously unobserved vibrationally resolved band, assigned to the B(2)A(") excited state of the furanyl radical, (?)C(4)H(3)O. The observed band origin corresponds to a 2.53 +/- 0.01 eV B(2)A(") term energy, while the resolved vibrational progression (853 +/- 42 cm(-1)) is assigned to an in-plane ring mode of alpha-(?)C(4)H(3)O (B(2)A(")). PMID- 21639440 TI - Nuclear motion on the orbitally degenerate electronic ground state of fully deuterated triatomic hydrogen. AB - Nuclear motion in the vicinity of conical intersections of the degenerate electronic ground state of fully deuterated triatomic hydrogen, D(3), is investigated with the aid of a time-dependent wavepacket approach in hyperspherical coordinates. Vibronic energy level spectra and the eigenfunctions are examined by including, for example, (1) geometric phase (GP) correction, (2) diagonal Born-Huang (BH) correction, and (3) both GP and BH corrections to the Born-Oppenheimer adiabatic Hamiltonian and finally by considering the nonadiabatic coupling between the two electronic surfaces explicitly. It emerges from this study that inclusion of both the GP and BH corrections is insufficient to explain the spectral features observed in the experiment. The latter are recovered by considering the complete two-states coupled Hamiltonian only. This study shows that both the GP and BH corrections constitute a minor part of the surface coupling effects, in particular, on the dynamics of the upper adiabatic sheet. Most importantly, we add that the experimental signature of the GP effect appears only in the observed shift of the eigenlevels of the electronic state when compared to those obtained from a completely Born-Oppenheimer Hamiltonian. The detail fine structure of the observed band of the electronic state is shaped by the off-diagonal derivative coupling elements of the nonadiabatic coupling operator. PMID- 21639441 TI - State-to-state differential and relative integral cross sections for rotationally inelastic scattering of H2O by hydrogen. AB - State-to-state differential cross sections (DCSs) for rotationally inelastic scattering of H(2)O by H(2) have been measured at 71.2 meV (574 cm(-1)) and 44.8 meV (361 cm(-1)) collision energy using crossed molecular beams combined with velocity map imaging. A molecular beam containing variable compositions of the (J = 0, 1, 2) rotational states of hydrogen collides with a molecular beam of argon seeded with water vapor that is cooled by supersonic expansion to its lowest para or ortho rotational levels (J(KaKc) = 0(00) and 1(01), respectively). Angular speed distributions of fully specified rotationally excited final states are obtained using velocity map imaging. Relative integral cross sections are obtained by integrating the DCSs taken with the same experimental conditions. Experimental state-specific DCSs are compared with predictions from fully quantum scattering calculations on the most complete H(2)O-H(2) potential energy surface. Comparison of relative total cross sections and state-specific DCSs show excellent agreement with theory in almost all details. PMID- 21639442 TI - Electron scattering in Pt(PF3)4: elastic scattering, vibrational, and electronic excitation. AB - Experimental absolute differential cross sections for elastic scattering, and for vibrational and electronic excitation of Pt(PF(3))(4) by low-energy electrons are presented. The elastic cross sections have a deep angle-dependent Ramsauer Townsend minimum (E(min) = 0.26 eV at theta = 135 degrees ). The angular distributions of the elastic cross section at and above 6.5 eV show an unusually narrow peak at an angle which decreases with increasing energy (it is at 40 degrees at 20 eV). Wavy structure is observed at higher angles at 15 and 20 eV. Vibrational excitation cross sections reveal five shape resonances, at 0.84, 1.75, 3.3, 6.6, and 8.5 eV. The angular distributions of the vibrational cross sections have a strong forward peak and are nearly isotropic above about 60 degrees . Electronically excited states are characterized by electron energy-loss spectra. They show a number of unstructured bands, the lowest at 5.8 eV. They are assigned to Rydberg states converging to the 1st and 2nd ionization energies. The cross sections for electronic excitation have very high forward peaks, reaching the value of 50 A(2) at 50 eV and 0 degrees scattering angle. Purity of the sample was monitored by the very low frequency (26 meV) Pt-P stretch vibration in the energy-loss spectra. PMID- 21639443 TI - Cl2O photochemistry: ultraviolet/vis absorption spectrum temperature dependence and O(3P) quantum yield at 193 and 248 nm. AB - The photochemistry of Cl(2)O (dichlorine monoxide) was studied using measurements of its UV/vis absorption spectrum temperature dependence and the O((3)P) atom quantum yield, Phi(Cl(2)O)(O)(lambda), in its photolysis at 193 and 248 nm. The Cl(2)O UV/vis absorption spectrum was measured over the temperature range 201-296 K between 200 and 500 nm using diode array spectroscopy. Cl(2)O absorption cross sections, sigma(Cl(2)O)(lambda,T), at temperatures <296 K were determined relative to its well established room temperature values. A wavelength and temperature dependent parameterization of the Cl(2)O spectrum using the sum of six Gaussian functions, which empirically represent transitions from the ground (1)A(1) electronic state to excited states, is presented. The Gaussian functions are found to correlate well with published theoretically calculated vertical excitation energies. O((3)P) quantum yields in the photolysis of Cl(2)O at 193 and 248 nm were measured using pulsed laser photolysis combined with atomic resonance fluorescence detection of O((3)P) atoms. O((3)P) quantum yields were measured to be 0.85 +/- 0.15 for 193 nm photolysis at 296 K and 0.20 +/- 0.03 at 248 nm, which was also found to be independent of temperature (220-352 K) and pressure (17 and 28 Torr, N(2)). The quoted uncertainties are at the 2sigma (95% confidence) level and include estimated systematic errors. ClO radical temporal profiles obtained following the photolysis of Cl(2)O at 248 nm, as reported previously in Feierabend et al. [J. Phys. Chem. A 114, 12052, (2010)], were interpreted to establish a <5% upper-limit for the O + Cl(2) photodissociation channel, which indicates that O((3)P) is primarily formed in the three-body, O + 2Cl, photodissociation channel at 248 nm. The analysis also indirectly provided a Cl atom quantum yield of 1.2 +/- 0.1 at 248 nm. The results from this work are compared with previous studies where possible. PMID- 21639444 TI - Reduced dimensionality spin-orbit dynamics of CH3 + HCl ? CH4 + Cl on ab initio surfaces. AB - A reduced dimensionality quantum scattering method is extended to the study of spin-orbit nonadiabatic transitions in the CH(3) + HCl ? CH(4) + Cl((2)P(J)) reaction. Three two-dimensional potential energy surfaces are developed by fitting a 29 parameter double-Morse function to CCSD(T)/IB//MP2/cc-pV(T+d)Z-dk ab initio data; interaction between surfaces is described by geometry-dependent spin orbit coupling functions fit to MCSCF/cc-pV(T+d)Z-dk ab initio data. Spectator modes are treated adiabatically via inclusion of curvilinear projected frequencies. The total scattering wave function is expanded in a vibronic basis set and close-coupled equations are solved via R-matrix propagation. Ground state thermal rate constants for forward and reverse reactions agree well with experiment. Multi-surface reaction probabilities, integral cross sections, and initial-state selected branching ratios all highlight the importance of vibrational energy in mediating nonadiabatic transition. Electronically excited state dynamics are seen to play a small but significant role as consistent with experimental conclusions. PMID- 21639445 TI - Vibrational and rotational cooling of NO+ in collisions with He. AB - A quantum mechanical investigation of the vibrational and rotational deactivation of NO(+) in collisions with He atoms in the cold and ultracold regime is presented. Ab initio potential energy calculations are carried out at BCCD(T) level and a new global 3D potential energy surface (PES) is obtained by fitting ab initio points within the reproducing kernel Hilbert space method. As a first test of this PES the bound state energies of the (3)He-NO(+) and (4)He-NO(+) complexes are calculated and compared to previous rigid rotor calculations. The efficiency of the vibrational and the rotational cooling of this molecular ion using a buffer gas of helium is then investigated by performing close coupling scattering calculations for collision energy ranging from 10(-6) to 2000 cm(-1). The calculations are performed for the two isotopes (3)He and (4)He and the results are compared to the available experimental data. PMID- 21639446 TI - Molecular dynamics simulations of nucleation from vapor to solid composed of Lennard-Jones molecules. AB - We performed molecular dynamics (MD) simulations of nucleation from vapor at temperatures below the triple point for systems consisting of 10(4)-10(5) Lennard Jones (L-J) type molecules in order to test nucleation theories at relatively low temperatures. Simulations are performed for a wide range of initial supersaturation ratio (S(0) ? 10-10(8)) and temperature (kT = 0.2-0.6epsilon), where epsilon and k are the depth of the L-J potential and the Boltzmann constant, respectively. Clusters are nucleated as supercooled liquid droplets because of their small size. Crystallization of the supercooled liquid nuclei is observed after their growth slows. The classical nucleation theory (CNT) significantly underestimates the nucleation rates (or the number density of critical clusters) in the low-T region. The semi-phenomenological (SP) model, which corrects the CNT prediction of the formation energy of clusters using the second virial coefficient of a vapor, reproduces the nucleation rate and the cluster size distributions with good accuracy in the low-T region, as well as in the higher-T cases considered in our previous study. The sticking probability of vapor molecules onto the clusters is also obtained in the present MD simulations. Using the obtained values of sticking probability in the SP model, we can further refine the accuracy of the SP model. PMID- 21639447 TI - Relativistic effects on the nuclear magnetic resonance shielding of FX (X = F, Cl, Br, I, and At) molecular systems. AB - We present ab inito full four-component and spin-free calculations of the NMR shielding parameter, sigma, in the FX (X = F, Cl, Br, I and At) molecular systems. A different expression that overcomes the traditional non-relativistic (NR) approximation used to calculate the relationship between spin-rotation constants and the paramagnetic terms of sigma(p) are given. Large deviations from NR results are obtained for sigma(X; X = I and At) and for sigma(F; FAt). sigma(?)(p)(I; FI) is zero within the NR approach but -447.4 parts per million from our calculations. The electronic origin of relativistic corrections are analyzed. All passive SO contributions are obtained as a difference between full four-component calculations and spin-free ones. Considering relativistic effects on the anisotropy, we obtain a deviation of 10% for I and 25% for At. sigma(?)(SO)(X) is always negative and sigma(?)(SF)(X) is always positive; the passive SO becomes larger than the SF one for X = Br, I, and At. Both sigma(?)(SO)(X) and sigma(?)(SO)(X) have a functional dependence such as a Z(X)(b) being the exponent 3.5 and 3.65, respectively. The passive SO contribution to the anisotropy has a similar functional dependence with an exponent of 3.60, meaning that its perpendicular component is larger than its corresponding parallel component. PMID- 21639448 TI - Structure and hydration of the C4H4?+ ion formed by electron impact ionization of acetylene clusters. AB - Here we report ion mobility experiments and theoretical studies aimed at elucidating the identity of the acetylene dimer cation and its hydrated structures. The mobility measurement indicates the presence of more than one isomer for the C(4)H(4)(?+) ion in the cluster beam. The measured average collision cross section of the C(4)H(4)(?+) isomers in helium (38.9 +/- 1 A(2)) is consistent with the calculated cross sections of the four most stable covalent structures calculated for the C(4)H(4)(?+) ion [methylenecyclopropene (39.9 A(2)), 1,2,3-butatriene (41.1 A(2)), cyclobutadiene (38.6 A(2)), and vinyl acetylene (41.1 A(2))]. However, none of the single isomers is able to reproduce the experimental arrival time distribution of the C(4)H(4)(?+) ion. Combinations of cyclobutadiene and vinyl acetylene isomers show excellent agreement with the experimental mobility profile and the measured collision cross section. The fragment ions obtained by the dissociation of the C(4)H(4)(?+) ion are consistent with the cyclobutadiene structure in agreement with the vibrational predissociation spectrum of the acetylene dimer cation (C(2)H(2))(2)(?+) [R. A. Relph, J. C. Bopp, J. R. Roscioli, and M. A. Johnson, J. Chem. Phys. 131, 114305 (2009)]. The stepwise hydration experiments show that dissociative proton transfer reactions occur within the C(4)H(4)(?+)(H(2)O)(n) clusters with n >= 3 resulting in the formation of protonated water clusters. The measured binding energy of the C(4)H(4)(?+)H(2)O cluster, 38.7 +/- 4 kJ/mol, is in excellent agreement with the G3(MP2) calculated binding energy of cyclobutadiene(?+).H(2)O cluster (41 kJ/mol). The binding energies of the C(4)H(4)(?+)(H(2)O)(n) clusters change little from n = 1 to 5 (39-48 kJ/mol) suggesting the presence of multiple binding sites with comparable energies for the water-C(4)H(4)(?+) and water-water interactions. A significant entropy loss is measured for the addition of the fifth water molecule suggesting a structure with restrained water molecules, probably a cyclic water pentamer within the C(4)H(4)(?+)(H(2)O)(5) cluster. Consequently, a drop in the binding energy of the sixth water molecule is observed suggesting a structure in which the sixth water molecule interacts weakly with the C(4)H(4)(?+)(H(2)O)(5) cluster presumably consisting of a cyclobutadiene(?+) cation hydrogen bonded to a cyclic water pentamer. The combination of ion mobility, dissociation, and hydration experiments in conjunction with the theoretical calculations provides strong evidence that the (C(2)H(2))(2)(?+) ions are predominantly present as the cyclobutadiene cation with some contribution from the vinyl acetylene cation. PMID- 21639449 TI - Nuclear spin selective alignment of ethylene and analogues. AB - We investigate the alignment of ethylene and of some of its analogues via short, non-resonant laser pulses and show that it depends crucially on the nuclear spin of the molecules. We calculate the time-dependent alignment factors of the four nuclear spin isomers of ethylene and analyze them by comparison with the symmetric top molecule allene. Moreover, we explore how the nuclear spin selective alignment depends on the asymmetry of the molecules and on the intensity of the laser pulse. As an application, we discuss how nuclear spin selective alignment could be applied in order to separate different isotopomers of ethylene. PMID- 21639450 TI - First-principles calculations of the structural and dynamic properties, and the equation of state of crystalline iodine oxides I2O4, I2O5, and I2O6. AB - The structural and dynamical correlations, and the equation of state of crystalline I(2)O(4), I(2)O(5), and I(2)O(6) are investigated by first-principles calculations based on the density functional theory (DFT). The lattice dynamics results reveal distinctive features in the phonon density of states among the three crystals. The frequencies of the stretch modes in I(2)O(4) and I(2)O(5) are clearly separated from those of the other (e.g., bending) modes by a gap, with all stretch modes above the gap. In contrast, the gap in I(2)O(6) separates the highest-frequency stretch modes with other stretch modes, and there is no gap between the stretch and the other modes in I(2)O(6). The motion of iodine atoms is involved in all vibrational modes in I(2)O(5), but only in low-frequency lattice modes in I(2)O(6). In I(2)O(4), iodine atoms are involved in modes with frequency below 700 cm(-1). Van der Waals correction within our DFT calculations is found to reduce the overestimation of the equilibrium volume, with its effect on structure similar to the pressure effect. Namely, both effects significantly decrease the inter-molecular distances, while slightly increasing the bond lengths within the molecules. This causes the frequencies of some vibrational modes to decrease with pressure, resulting in negative "modes Gruneisen parameters" for those modes. Thermodynamic properties, derived from the equation of state, of crystalline I(2)O(4), I(2)O(5), and I(2)O(6) are discussed within the quasi-harmonic approximation. PMID- 21639451 TI - Structural heterogeneity and pressure-relaxation in compressed borosilicate glasses by in situ small angle X-ray scattering. AB - We report on Brillouin and in situ small angle X-ray scattering (SAXS) analyses of topological heterogeneity in compressed sodium borosilicate glasses. SAXS intensity extrapolated to very low angular regimes, I(q = 0), is related to compressibility. From Brillouin scattering and analyses of the elastic properties of the glass, the Landau-Placzek ratio is determined and taken as a direct reflection of the amplitude of frozen-in density fluctuations. It is demonstrated that with increasing fictive pressure, topological (mid- and long-range) homogeneity of the glass increases significantly. Heating and cooling as well as isothermal scans were performed to follow the evolution of density fluctuations upon pressure recovery. For a sample with a fictive pressure p(f) of 470 MPa, complete recovery to p(f) = 0.1 MPa was observed to occur close to the glass transition temperature. The values of fictive and apparent fictive temperature, respectively, as obtained via the intersection method from plots of I(q = 0) vs. temperature were found in good agreement with previous calorimetric analyses. Isothermal scans suggest that mid- and long-range recovery govern macroscopic density relaxation. PMID- 21639452 TI - Simulations of a lattice model of two-headed linear amphiphiles: influence of amphiphile asymmetry. AB - Using a 2D lattice model, we conduct Monte Carlo simulations of micellar aggregation of linear-chain amphiphiles having two solvophilic head groups. In the context of this simple model, we quantify how the amphiphile architecture influences the critical micelle concentration (CMC), with a particular focus on the role of the asymmetry of the amphiphile structure. Accordingly, we study all possible arrangements of the head groups along amphiphile chains of fixed length N = 12 and 16 molecular units. This set of idealized amphiphile architectures approximates many cases of symmetric and asymmetric gemini surfactants, double headed surfactants, and boloform surfactants. Consistent with earlier results, we find that the number of spacer units s separating the heads has a significant influence on the CMC, with the CMC increasing with s for s < N/2. In comparison, the influence of the asymmetry of the chain architecture on the CMC is much weaker, as is also found experimentally. PMID- 21639453 TI - Structure and reactivity of pyridine crystal under pressure. AB - In this work we have performed an extensive high pressure study of the condensed phases of pyridine by Raman and IR spectroscopy. We have evidenced three different polymorphs, two crystalline, and one glassy and established the pressure conditions in which they exist as stable or metastable phases by several compression/decompression experiments both on annealed and not annealed samples. Crystallization and phase transitions are found to be kinetically driven. The vibrational spectra are extremely complex due to the low symmetry of the crystals, which implies a large number of crystal components. This complexity required a careful analysis of both IR and Raman data that led to the identification of 20 out of 21 external modes expected for phase II. We did not find any conclusive indication of phase transitions on compressing phase II thus indicating that phase II is likely the stable phase at the onset pressure of the chemical transformation of pyridine. The latter starts at 18 GPa and relevant differences from the well characterized benzene reaction suggest that it is likely driven by crystal defects. PMID- 21639454 TI - Two-dimensional infrared spectroscopy of isotope-diluted ice Ih. AB - We present experimental 2D IR spectra of isotope diluted ice Ih (i.e., the OH stretch mode of HOD in D(2)O and the OD stretch mode of HOD in H(2)O) at T = 80 K. The main spectral features are the extremely broad 1-2 excited state transition, much broader than the corresponding 0-1 groundstate transition, as well as the presence of quantum beats. We do not observe any inhomogeneous broadening that might be expected due to proton disorder in ice Ih. Complementary, we perform simulations in the framework of the Lippincott Schroeder model, which qualitatively reproduce the experimental observations. We conclude that the origin of the observed line shape features is the coupling of the OH-vibrational coordinate with crystal phonons and explain the beatings as a coherent oscillation of the O...O hydrogen bond degree of freedom. PMID- 21639455 TI - A powerful computational crystallography method to study ice polymorphism. AB - Classical molecular dynamics (MD) simulations are employed as a tool to investigate structural properties of ice crystals under several temperature and pressure conditions. All ice crystal phases are analyzed by means of a computational protocol based on a clustering approach following standard MD simulations. The MD simulations are performed by using a recently published classical interaction potential for oxygen and hydrogen in bulk water, derived from neutron scattering data, able to successfully describe complex phenomena such as proton hopping and bond formation/breaking. The present study demonstrates the ability of the interaction potential model to well describe most ice structures found in the phase diagram of water and to estimate the relative stability of 16 known phases through a cluster analysis of simulated powder diagrams of polymorphs obtained from MD simulations. The proposed computational protocol is suited for automated crystal structure identification. PMID- 21639456 TI - Liquid-liquid phase transformations and the shape of the melting curve. AB - The phase diagram of elemental liquids has been found to be surprisingly rich, including variations in the melting curve and transitions in the liquid phase. The effect of these transitions in the liquid state on the shape of the melting curve is analyzed. First-order phase transitions intersecting the melting curve imply piecewise continuous melting curves, with solid-solid transitions generating upward kinks or minima and liquid-liquid transitions generating downward kinks or maxima. For liquid-liquid phase transitions proposed for carbon, phosphorous selenium, and possibly nitrogen, we find that the melting curve exhibits a kink. Continuous transitions imply smooth extrema in the melting curve, the curvature of which is described by an exact thermodynamic relation. This expression indicates that a minimum in the melting curve requires the solid compressibility to be greater than that of the liquid, a very unusual situation. This relation is employed to predict the loci of smooth maxima at negative pressures for liquids with anomalous melting curves. The relation between the location of the melting curve maximum and the two-state model of continuous liquid-liquid transitions is discussed and illustrated by the case of tellurium. PMID- 21639457 TI - Impact of intramolecular twisting and exciton migration on emission efficiency of multifunctional fluorene-benzothiadiazole-carbazole compounds. AB - Novel donor-acceptor compounds consisting of singly bonded fluorene (Fl), benzothiadiazole (BT), and carbazole (Cz) functional units in the same molecule were investigated. Analysis of the optical spectra and fluorescence transients of the compounds revealed the domination of intramolecular charge transfer (ICT) states with high fluorescence quantum yield (72%-85%). A similar Cz-Fl-Cz compound exhibiting 100% fluorescence quantum yield and no ICT character was also studied as a reference to reveal the impact of electron-accepting BT groups. Thorough examination of the optical properties of the compounds in different media, i.e., dilute solution and polymer matrix, indicated their twisted conformations due to steric hindrance in the ground state and flattened geometry in the excited state for both reference and ICT compounds. Remarkable fluorescence efficiency losses (amounting to 70%) observed upon casting the molecular solutions into neat films were determined to originate from the low fluorescent twisted conformers and migration-facilitated exciton quenching. The majority of emission efficiency losses (over 70%) were caused by the twisted conformers, whereas only less than 30% by exciton-migration-induced nonradiative deactivation. PMID- 21639458 TI - Vibrational and thermodynamic properties of beta-HMX: a first-principles investigation. AB - Thermodynamic properties of beta-HMX crystal are investigated using the quasi harmonic approximation and density functional theory within the local density approximation (LDA), generalized gradient approximation (GGA), and GGA + empirical van der Waals (vdW) correction. It is found that GGA well describes the thermal expansion coefficient and heat capacity but fails to produce correct bulk modulus and equilibrium volume. The vdW correction improves the bulk modulus and volume, but worsens the thermal expansion coefficient and heat capacity. In contrast, LDA describes all thermodynamic properties with reasonable accuracy, and overall is a good exchange-correlation functional for beta-HMX molecular crystal. The results also demonstrate significant contributions of phonons to the equation of state. The static calculation of equilibrium volume for beta-HMX differs from the room-temperature value incorporating lattice vibrations by over 5%. Therefore, for molecular crystals, it is essential to include phonon contributions when calculated equation of state is compared with experimental data at ambient condition. PMID- 21639459 TI - Ab initio calculation of the potential of mean force for dissociation of aqueous Ca-Cl. AB - The potential of mean force for the dissociation of a Ca-Cl ion pair in water is calculated from ab initio molecular dynamics simulations. The constraint-force method is employed to enhance sampling over the entire range of the reaction coordinate (Ca-Cl distance) from 2.2 to 6.5 A. Particular attention is paid to equilibration of the system as it is found that the potential of mean force is highly sensitive to the hydration number of the Ca(2+) ion. The structure and polarization of hydration waters are examined in detail at three ion-ion separations of interest: the contact-ion position, the solvent-separated-ion position, and the transition state between them. The ab initio results are compared to the classical ones obtained using the CHARMM force field and the parameters of Dang and Smith. There are substantial differences between the polarization of hydration waters of Ca(2+) and Cl(-) ions at all distances, which indicates that an accurate description of Ca-Cl dissociation with nonpolarizable force fields may not be feasible. The ab initio results presented here for the Ca Cl ion pair complements our earlier results for Na-Cl, and together they provide useful benchmarks for polarizable force fields under construction. PMID- 21639460 TI - Vibrational spectroscopy and relaxation of an anharmonic oscillator coupled to harmonic bath. AB - The vibrational spectroscopy and relaxation of an anharmonic oscillator coupled to a harmonic bath are examined to assess the applicability of the time correlation function (TCF), the response function, and the semiclassical frequency modulation (SFM) model to the calculation of infrared (IR) spectra. These three approaches are often used in connection with the molecular dynamics simulations but have not been compared in detail. We also analyze the vibrational energy relaxation (VER), which determines the line shape and is itself a pivotal process in energy transport. The IR spectra and VER are calculated using the generalized Langevin equation (GLE), the Gaussian wavepacket (GWP) method, and the quantum master equation (QME). By calculating the vibrational frequency TCF, a detailed analysis of the frequency fluctuation and correlation time of the model is provided. The peak amplitude and width in the IR spectra calculated by the GLE with the harmonic quantum correction are shown to agree well with those by the QME though the vibrational frequency is generally overestimated. The GWP method improves the peak position by considering the zero-point energy and the anharmonicity although the red-shift slightly overshoots the QME reference. The GWP also yields an extra peak in the higher-frequency region than the fundamental transition arising from the difference frequency of the center and width oscillations of a wavepacket. The SFM approach underestimates the peak amplitude of the IR spectra but well reproduces the peak width. Further, the dependence of the VER rate on the strength of an excitation pulse is discussed. PMID- 21639461 TI - Transient switch-on/off currents in molecular junctions. AB - Based on the nonequilibrium density matrix theory we put forward a unified description of the transient and the steady state current formation through a molecular junction. It is demonstrated that the current follows the time evolution of the populations of those molecular charged states which participate in the inter-electrode charge transmission. As an example, the formation of switch-on/switch-off currents is analyzed for a junction where the molecule has two active terminal sites. It is shown that just after a sudden voltage switch-on or switch-off, the resulting transient currents can significantly exceed their steady state value. This feature is caused by molecular charging or discharging processes, which are fast compared to those processes responsible for establishing the steady state current in the junction. The largest transient currents appear if the coupling of the molecule to the adjacent electrodes is asymmetric, or if the applied voltage causes a transformation of extended molecular states into localized ones. PMID- 21639462 TI - A unique vibrational signature of rotated water monolayers on Pt(111): predicted and observed. AB - Six H-bonds in the periodic di-interstitial structure that accounts for scanning tunneling microscope images of "?37" and "?39" wetting layers on Pt(111) are some 0.2 A shorter than H-bonds are in ice Ih. According to a broadly obeyed correlation, this density functional theory result implies a stringent test of the di-interstitial motif, namely the presence of an OH-stretch band red-shifted from that of ice Ih by more than 1000 cm(-1). Infrared absorption spectra satisfy the test, in showing a feature centered at about 1965 cm(-1), which grows in as deposited water orders. PMID- 21639463 TI - The nature of singlet excitons in oligoacene molecular crystals. AB - A theory for polarized absorption in crystalline oligoacenes is presented, which includes Frenkel exciton coupling, the coupling between Frenkel and charge transfer (CT) excitons, and the coupling of all neutral and ionic excited states to the dominant ring-breathing vibrational mode. For tetracene, spectra calculated using all Frenkel couplings among the five lowest energy molecular singlet states predict a Davydov splitting (DS) of the lowest energy (0-0) vibronic band of only -32 cm(-1), far smaller than the measured value of 631 cm( 1) and of the wrong sign-a negative sign indicating that the polarizations of the lower and upper Davydov components are reversed from experiment. Inclusion of Frenkel-CT coupling dramatically improves the agreement with experiment, yielding a 0-0 DS of 601 cm(-1) and a nearly quantitative reproduction of the relative spectral intensities of the 0-n vibronic components. Our analysis also shows that CT mixing increases with the size of the oligoacenes. We discuss the implications of these results on exciton dissociation and transport. PMID- 21639464 TI - Synchrotron x-ray photoemission study of soft x-ray processed ultrathin glycine water ice films. AB - Ultrathin glycine-water ice films have been prepared in ultrahigh vacuum by condensation of H(2)O and glycine at 90 K on single crystalline alumina surfaces and processed by soft x-ray (610 eV) exposure for up to 60 min. The physicochemical changes in the films were monitored using synchrotron x-ray photoemission spectroscopy. Two films with different amounts of H(2)O have been considered in order to evaluate the influence of the water ice content on the radiation-induced effects. The analysis of C1s, N1s, and O1s spectral regions together with the changes in the valence band spectra indicates that amino acid degradation occurs fast mainly via decarboxylation and deamination of pristine molecules. Enrichment of the x-ray exposed surfaces with fragments with carbon atoms without strong electronegative substituents (C-C and C-H) is documented as well. In the thinner glycine-water ice film (six layers of glycine + six layers of water) the 3D ice suffers strongly from the x-rays and is largely removed from the sample. The rate of photodecomposition of glycine in this film is about 30% higher than for glycine in the thicker film (6 layers of glycine + 60 layers of water). The photoemission results suggest that the destruction of amino acid molecules is caused by the direct interaction with the radiation and that no chemical attack of glycine by the species released by water radiolysis is detected. PMID- 21639465 TI - Structural and electronic properties of ZrX2)and HfX2 (X=S and Se) from first principles calculations. AB - Early transition metal dichalcogenides (TMDC), characterized by their quasi-two dimensional layered structure, have attracted intensive interest due to their versatile chemical and physical properties, but a comprehensive understanding of their structural and electronic properties from a first-principles point of view is still lacking. In this work, four simple TMDC materials, MX(2) (M = Zr and Hf, X = S and Se), are investigated by the Kohn-Sham density functional theory (KS DFT) with different local or semilocal exchange-correlation (xc) functionals and many-body perturbation theory in the GW approximation. Although the widely used Perdew-Burke-Ernzelhof (PBE) generalized gradient approximation (GGA) xc functional overestimates the interlayer distance dramatically, two newly developed GGA functionals, PBE-for-solids (PBEsol) and Wu-Cohen 2006 (WC06), can reproduce experimental crystal structures of these TMDC materials very well. The GW method, currently the most accurate first-principles approach for electronic band structures of extended systems, gives the fundamental band gaps of all these materials in good agreement with the experimental values obtained from optical absorption. The minimal direct gaps from GW are systematically larger than those measured from thermoreflectance by about 0.1-0.3 eV, implying that excitonic effects may be stronger than previously estimated. The calculated density of states from GW quasi-particle band energies agrees very well with photo-emission spectroscopy data. Ionization potentials of these materials are also computed by combining PBE calculations based on the slab model and GW quasi-particle corrections. The calculated absolute band energies with respect to the vacuum level indicate that that ZrS(2) and HfS(2), although having suitable band gaps for visible light absorption, cannot be used for overall water splitting as a result of mismatch of the conduction band minimum with the redox potential of H(+)/H(2). PMID- 21639466 TI - Effects of electric field on confined electrolyte in a hexagonal mesoporous silica. AB - In an electrowetting experiment on a surface treated hexagonal mesoporous silica, it is noticed that the effective solid-liquid interfacial tension is quite insensitive to the applied voltage, while the accessible nanopore volume decreases significantly as the voltage is increased. When the voltage is higher than 900 V, the liquid infiltration cannot be detected. The liquid defiltration is quite insensitive to the electric field. These unique phenomena may be attributed to the field responsive ion behaviors in the confining nanoenvironment. PMID- 21639467 TI - Unoccupied states in Cu and Zn octaethyl-porphyrin and phthalocyanine. AB - Copper and zinc phthalocyanines and porphyrins are used in organic light emitting diodes and dye-sensitized solar cells. Using near edge x-ray absorption fine structure (NEXAFS) spectroscopy at the Cu 2p and Zn 2p edges, the unoccupied valence states at the Cu and Zn atoms are probed and decomposed into 3d and 4s contributions with the help of density functional calculations. A comparison with the N 1s edge provides the 2p states of the N atoms surrounding the metal, and a comparison with inverse photoemission provides a combined density of states. PMID- 21639468 TI - Very-high-strength (60-GPa) carbon nanotube fiber design based on molecular dynamics simulations. AB - The mechanical properties of carbon nanotubes such as low density, high stiffness, and exceptional strength make them ideal candidates for reinforcement material in a wide range of high-performance composites. Molecular dynamics simulations are used to predict the tensile response of fibers composed of aligned carbon nanotubes with intermolecular bonds of interstitial carbon atoms. The effects of bond density and carbon nanotube length distribution on fiber strength and stiffness are investigated. The interstitial carbon bonds significantly increase load transfer between the carbon nanotubes over that obtained with van der Waals forces. The simulation results indicate that fibers with tensile strengths to 60 GPa could be produced by employing interstitial cross-link atoms. The elastic modulus of the fibers is also increased by the bonds. PMID- 21639469 TI - A volume averaging approach for asymmetric diffusion in porous media. AB - Asymmetric diffusion has been observed in different contexts, from transport in stratified and fractured porous media to diffusion of ions and macromolecular solutes through channels in biological membranes. Experimental and numerical observations have shown that diffusion is facilitated in the direction of positive void fraction (i.e., porosity) gradients. This work uses the method of volume averaging in order to obtain effective medium equations for systems with void fraction gradients for passive and diffusive mass transport processes. The effective diffusivity is computed from the solution of an associated closure problem in representative unit cells that allow considering porosity gradients. In this way, the results in this work corroborate previous findings showing that the effective diffusivity exhibits important directional asymmetries for geometries with void fraction gradients. Numerical examples for simple geometries (a section with an obstacle and a channel with varying cross section) show that the diffusion asymmetry depends strongly on the system configuration. The magnitude of this dependence can be quantified from the results in this work. PMID- 21639470 TI - Effect of co-adsorbed CO and reaction temperature on the dynamics of N2 desorption under steady-state N2O-CO reaction on Rh(110). AB - We have investigated the effect of co-absorbed CO and reaction temperature on the angular distribution of N(2) desorption by N(2)O decomposition under the steady state of N(2)O-CO reaction on Rh(110). Spatial distributions of desorbing product N(2) emission have been measured at various surface temperatures and CO coverages. The decomposed N(2) collimates at 48 degrees -61 degrees off normal in the parallel plane to [001] and [110] directions, indicating that adsorbed N(2)O just before the decomposition is oriented along the [001] direction. Although the inclined and collimated N(2) desorption is always observed at any steady-state CO coverage and reaction temperature, the shape of the collimated N(2) distribution varied dependent on the co-adsorbed CO coverage. The distribution becomes sharp and shifts toward the surface normal direction with increasing CO coverage. These effects of adsorbed CO on the angular distribution of N(2) are interpreted by the collision of desorbed N(2) with co-adsorbed CO. PMID- 21639471 TI - Influence of the block hydrophilicity of AB2 miktoarm star copolymers on cluster formation in solutions. AB - We investigated the formation of various micelle shapes of lipid-like amphiphilic AB(2) miktoarm star copolymers in a solution, by performing dissipative particle dynamics simulations. AB(2) miktoarm star copolymer molecules are modeled with coarse-grained structures that consist of a relatively hydrophilic head (A) group with a single arm and a hydrophobic tail (B) group with double arms. A decrease in the hydrophilicity of the head group leads to a reduction of the polymer solvent contact area, causing cluster structure changes from spherical micelles to vesicles. Consequently, a spherical exterior with multi-lamellar or cylindrical phase interior structures forms under poor solvent conditions without the introduction of spherical hard-wall containers. Furthermore we observed that, for small head group lengths, vesicles were formed in much wider range of solvent head interaction strength than for long head groups, indicating that molecules with short head group offer a superior vesicle forming property. A phase diagram, the structure and kinetics of the cluster formation, a density profile, and a detailed shape analysis are presented to discuss the molecular characteristics of potential candidates for drug carriers that require superior and versatile vesicle forming properties. We also show that, under certain solvent-hydrophilic head group interaction conditions, initially formed cylindrical micelles transform to bilayer fragments through redistribution of copolymers within the cluster. PMID- 21639472 TI - Monte Carlo simulations of charged dendrimer-linear polyelectrolyte complexes and explicit counterions. AB - We study complexes composed of one dendrimer of generation G = 4 (G4 dendrimer) with N(t) = 32 charged terminal groups and an oppositely charged linear polyelectrolyte accompanied by neutralizing counterions in an athermal solvent using Monte Carlo simulations based on the bond fluctuation model. In our study both the full Coulomb potential and the excluded volume interactions are taken into account explicitly with the reduced temperature tau and the chain length N(ch) as the main simulation parameters. Our calculations indicate that there exist three temperature ranges that determine the behavior of such complexes. At tau(complex) stable charged dendrimer-linear polyelectrolyte complexes are formed first, which are subsequently accompanied by selective counterion localization within the complex interior at tau(loc) <= tau(complex), and counterion condensation as temperature is further decreased below tau(cond) < tau(loc). In particular, we observe that condensation takes place exclusively on the excess charges in the complex and thus no condensation is observed at the compensation point (N(ch) = N(t)), irrespective of tau. For N(ch) ? N(t) the complex is overally charged. Furthermore, we discuss the size and structure of the dendrimer and the linear polyelectrolyte within the complex, as well as spatial distributions of monomers and counterions. Conformations of the chain in the bound state are analysed in terms of loops, trains, and tails. PMID- 21639473 TI - Nanostructures in a binary mixture confined in slit-like pores with walls decorated with tethered polymer brushes in the form of stripes: dissipative particle dynamics study. AB - Using dissipative particle dynamics, we investigate the behavior of a binary mixture, exhibiting demixing in a bulk phase, confined in slit-like pores with walls modified by the stripes of tethered brush of chains. Our main interest is to determine possible morphologies that can be formed inside the pore, depending on the geometrical parameters characterizing the system (the size of the pore and the width of the stripes). In order to describe the observed morphologies we calculate several characteristics, as the density and local temperature profiles, the radii of gyration for the attached polymers, and the minimum polymer-polymer distances in the direction parallel and perpendicular to the pore walls. The summary of our findings is presented as a sketch of the diagram of morphologies. PMID- 21639474 TI - Molecular dynamics simulation study of nonconcatenated ring polymers in a melt. I. Statics. AB - Molecular dynamics simulations were conducted to investigate the structural properties of melts of nonconcatenated ring polymers and compared to melts of linear polymers. The longest rings were composed of N = 1600 monomers per chain which corresponds to roughly 57 entanglement lengths for comparable linear polymers. For the rings, the radius of gyration squared, [linear span]R(g)(2)[linear span], was found to scale as N(4/5) for an intermediate regime and N(2/3) for the larger rings indicating an overall conformation of a crumpled globule. However, almost all beads of the rings are "surface beads" interacting with beads of other rings, a result also in agreement with a primitive path analysis performed in the next paper [J. D. Halverson, W. Lee, G. S. Grest, A. Y. Grosberg, and K. Kremer, J. Chem. Phys. 134, 204905 (2011)]. Details of the internal conformational properties of the ring and linear polymers as well as their packing are analyzed and compared to current theoretical models. PMID- 21639475 TI - Molecular dynamics simulation study of nonconcatenated ring polymers in a melt. II. Dynamics. AB - Molecular dynamics simulations were conducted to investigate the dynamic properties of melts of nonconcatenated ring polymers and compared to melts of linear polymers. The longest rings were composed of N = 1600 monomers per chain which corresponds to roughly 57 entanglement lengths for comparable linear polymers. The ring melts were found to diffuse faster than their linear counterparts, with both architectures approximately obeying a D ~ N(-2.4) scaling law for large N. The mean-square displacement of the center-of-mass of the rings follows a sub-diffusive behavior for times and distances beyond the ring extension [linear span]R(g)(2)[linear span], neither compatible with the Rouse nor the reptation model. The rings relax stress much faster than linear polymers, and the zero-shear viscosity was found to vary as eta(0) ~ N(1.4 +/- 0.2) which is much weaker than the N(3.4) behavior of linear chains, not matching any commonly known model for polymer dynamics when compared to the observed mean square displacements. These findings are discussed in view of the conformational properties of the rings presented in the preceding paper [J. D. Halverson, W. Lee, G. S. Grest, A. Y. Grosberg, and K. Kremer, J. Chem. Phys. 134, 204904 (2011)]. PMID- 21639476 TI - Quasielastic neutron scattering study of hydrogen motions in an aqueous poly(vinyl methyl ether) solution. AB - We present a quasielastic neutron scattering (QENS) investigation of the component dynamics in an aqueous Poly(vinyl methyl ether) (PVME) solution (30% water content in weight). In the glassy state, an important shift in the Boson peak of PVME is found upon hydration. At higher temperatures, the diffusive-like motions of the components take place with very different characteristic times, revealing a strong dynamic asymmetry that increases with decreasing T. For both components, we observe stretching of the scattering functions with respect to those in the bulk and non-Gaussian behavior in the whole momentum transfer range investigated. To explain these observations we invoke a distribution of mobilities for both components, probably originated from structural heterogeneities. The diffusive-like motion of PVME in solution takes place faster and apparently in a more continuous way than in bulk. We find that the T dependence of the characteristic relaxation time of water changes at T ? 225 K, near the temperature where a crossover from a low temperature Arrhenius to a high temperature cooperative behavior has been observed by broadband dielectric spectroscopy (BDS) [S. Cerveny, J. Colmenero and A. Alegria, Macromolecules, 38, 7056 (2005)]. This observation might be a signature of the onset of confined dynamics of water due to the freezing of the PVME dynamics, that has been selectively followed by these QENS experiments. On the other hand, revisiting the BDS results on this system we could identify an additional "fast" process that can be attributed to water motions coupled with PVME local relaxations that could strongly affect the QENS results. Both kinds of interpretations, confinement effects due to the increasing dynamic asymmetry and influence of localized motions, could provide alternative scenarios to the invoked "strong-to-fragile" transition. PMID- 21639477 TI - Fluid phase separation inside a static periodic field: an effectively two dimensional critical phenomenon. AB - When a fluid with a bulk liquid-vapor critical point is placed inside a static external field with spatial periodic oscillations in one direction, a new phase arises. This new phase-the so-called "zebra" phase-is characterized by an average density roughly between that of the liquid and vapor phases. The presence of the zebra phase gives rise to two new phase transitions: one from the vapor to the zebra phase, and one from the zebra to the liquid phase. At appropriate values of the temperature and chemical potential, the latter two transitions become critical. This phenomenon is called laser-induced condensation [I. O. Gotze, J. M. Brader, M. Schmidt, and H. Lowen, Mol. Phys. 101, 1651 (2003)]. The purpose of this paper is to elucidate the nature of the critical points, using density functional theory and computer simulation of a colloid-polymer mixture. The main finding is that critical correlations develop in two-dimensional sheets perpendicular to the field direction, but not in the direction along the field: the critical correlations are thus effectively two-dimensional. Hence, static periodic fields provide a means to confine a fluid to effectively two dimensions. Away from criticality, the vapor-zebra and liquid-zebra transitions become first order, but the associated surface tensions are extremely small. The consequences of the extremely small surface tensions on the nature of the two-phase coexistence regions are analyzed in detail. PMID- 21639478 TI - Glassy dynamics and mechanical response in dense fluids of soft repulsive spheres. I. Activated relaxation, kinetic vitrification, and fragility. AB - The microscopic nonlinear Langevin equation theory of activated glassy dynamics is applied to dense fluids of spherical particles that interact via a finite range Hertzian contact soft repulsion. The activation barrier and mean alpha relaxation time are predicted to be rich functions of volume fraction and particle stiffness, exhibiting a non-monotonic variation with concentration at high volume fractions. The latter is due to a structural "soft jamming" crossover where the real space local cage order weakens when soft particles significantly overlap. The highly variable dependences of the relaxation time on temperature and volume fraction are reasonably well collapsed onto two distinct master curves that are qualitatively consistent with a recent scaling ansatz and computer simulation study. A kinetic vitrification diagram is constructed and compared to its dynamic crossover analog. Intersection of the dynamic crossover and soft jamming threshold boundaries occurs for particles that are sufficiently soft, implying the nonexistence of a clear activated dynamics regime or kinetic arrest transition for such particles. The isothermal dynamic fragility is predicted to vary over a wide range as a function of particle stiffness, and soft particles behave as strong glasses. Qualitative comparisons with simulations and microgel experiments reveal good agreement. PMID- 21639479 TI - Glassy dynamics and mechanical response in dense fluids of soft repulsive spheres. II. Shear modulus, relaxation-elasticity connections, and rheology. AB - We apply the quiescent and mechanically driven versions of nonlinear Langevin equation theory to study how particle softness influences the shear modulus, the connection between shear elasticity and activated relaxation, and nonlinear rheology of the repulsive Hertzian contact model of dense soft sphere fluids. Below the soft jamming threshold, the shear modulus follows a power law dependence on volume fraction over a narrow interval with an apparent exponent that grows with particle stiffness. To a first approximation, the elastic modulus and transient localization length are controlled by a single coupling constant determined by local fluid structure. In contrast to the behavior of hard spheres, an approximately linear relation between the shear modulus and activation barrier is predicted. This connection has recently been observed for microgel suspensions and provides a microscopic realization of the elastic shoving model. Yielding, shear and stress thinning of the alpha relaxation time and viscosity, and flow curves are also studied. Yield strains are relatively weakly dependent on volume fraction and particle stiffness. Shear thinning commences at values of the effective Peclet number far less than unity, a signature of stress-assisted activated relaxation when barriers are high. Apparent power law reduction of the viscosity with shear rate is predicted with a thinning exponent less than unity. In the vicinity of the soft jamming threshold, a power law flow curve occurs over an intermediate reduced shear rate range with an apparent exponent that decreases as fluid volume fraction and/or repulsion strength increase. PMID- 21639480 TI - Estimating the viscoelastic moduli of complex fluids from observation of Brownian motion of a particle confined to a harmonic trap. AB - A procedure is proposed to estimate the viscoelastic properties of a complex fluid from the behavior of the velocity autocorrelation function of a suspended Brownian particle, trapped in a harmonic potential. The procedure is tested for a model complex fluid with a given frequency-dependent shear viscosity. The analysis shows that the procedure can provide a rather accurate prediction of the viscoelastic properties of the fluid on the basis of experimental data on the velocity autocorrelation function of the trapped Brownian particle in a limited range of time. PMID- 21639481 TI - Protein's electronic polarization contributes significantly to its catalytic function. AB - Ab initio quantum mechanical/molecular mechanical method is combined with the polarized protein-specific charge to study the chemical reactions catalyzed by protein enzymes. Significant improvement in the accuracy and efficiency of free energy simulation is demonstrated by calculating the free-energy profile of the primary proton transfer reaction in triosephosphate isomerase. Quantitative agreement with experimental results is achieved. Our simulation results indicate that electronic polarization makes important contribution to enzyme catalysis by lowering the energy barrier by as much as 3 kcal/mol. PMID- 21639482 TI - Combined effects of metal complexation and size expansion in the electronic structure of DNA base pairs. AB - Novel DNA derivatives have been recently investigated in the pursuit of modified DNA duplexes to tune the electronic structure of DNA-based assemblies for nanotechnology applications. Size-expanded DNAs (e.g., xDNA) and metalated DNAs (M-DNA) may enhance stacking interactions and induce metallic conductivity, respectively. Here we explore possible ways of tailoring the DNA electronic structure by combining the aromatic size expansion with the metal-doping. We select the salient structures from our recent study on natural DNA pairs complexed with transition metal ions and consider the equivalent model configurations for xDNA pairs. We present the results of density functional theory electronic structure calculations of the metalated expanded base-pairs with various localized basis sets and exchange-correlation functionals. Implicit solvent and coordination water molecules are also included. Our results indicate that the effect of base expansion is largest in Ag-xGC complexes, while Cu-xGC complexes are the most promising candidates for nanowires with enhanced electron transfer and also for on-purpose modification of the DNA double-helix for signal detection. PMID- 21639483 TI - Mixed quantum classical simulations of excitons in peptide helices. AB - We use mixed classical/quantum simulations to study the time dependence of an excitation of a C=O vibration on a 3-10 helix of alpha-aminoisobutyric acid, a system which represents a test case for the formation of self-trapped vibrational excitation states on protein helices. Due to the inherent disorder in the system caused by the finite temperature and fluctuations in hydrogen bonding, the excitation tunnels randomly among C=O sites along the helix. Quantum forces are insufficient to establish a coherent relationship between the location of the excitation and the contraction of hydrogen bonds around this site. Our simulations indicate that the excitation frequently becomes localized on the end of the helix due to the defect in helical structure caused by unwinding. Our results generally do not support the existence of Davydov type solitons in biological helix systems under physiological conditions. PMID- 21639484 TI - The protein folding network indicates that the ultrafast folding mutant of villin headpiece subdomain has a deeper folding funnel. AB - Protein folding is a dynamic process with continuous transitions among different conformations. In this work, the dynamics in the protein folding network of villin headpiece subdomain (HP35) has been investigated based on multiple reversible folding trajectories of HP35 and its ultrafast folding mutant where sub-angstrom folding was achieved. The four folding states were clearly separated on the network, validating the classification of the states. Examination of the eight conformers with different formation of the individual helices revealed high plasticity of the three helices in all the four states. A consistent feature between the wild type and mutant protein is the dominant conformer 111 (all three helices formed) in the folded state and conformers 111 and 011 (helices II and III formed) in the major intermediate state, indicating the critical role of helices II and III in the folding mechanism. When compared to the wild type, the folding landscape of the ultrafast folding mutant exhibited a deeper folding funnel towards the folded state. The very beginning of the folding (0-10 ns) was very similar for both protein variants but it soon diverged and displayed different folding pathways. Although going through the major intermediate state is the dominant pathway for both, it was also observed that some folding went through the minor intermediate state for the mutant. The intriguing difference resulting from the mutation at two residues in helix III has been carefully analyzed and discussed in details. PMID- 21639485 TI - Thermodynamics of DNA hybridization on surfaces. AB - Hybridization of single-stranded DNA (ssDNA) targets to surface-tethered ssDNA probes was simulated using an advanced coarse-grain model to identify key factors that influence the accuracy of DNA microarrays. Comparing behavior in the bulk and on the surface showed, contrary to previous assumptions, that hybridization on surfaces is more thermodynamically favorable than in the bulk. In addition, the effects of stretching or compressing the probe strand were investigated as a model system to test the hypothesis that improving surface hybridization will improve microarray performance. The results in this regard indicate that selectivity can be increased by reducing overall sensitivity by a small degree. Taken as a whole, the results suggest that current methods to enhance microarray performance by seeking to improve hybridization on the surface may not yield the desired outcomes. PMID- 21639486 TI - Characterizing plasma mirrors near breakdown. AB - Experiments dedicated to the characterization of plasma mirrors with a high energy, single shot short-pulse laser were performed at the 100 TW target area of the Z-Backlighter Facility at Sandia National Laboratories. A suite of beam diagnostics was used to characterize a high energy laser pulse with a large aperture through focus imaging setup. By varying the fluence on the plasma mirror around the plasma ignition threshold, critical performance parameters were determined and a more detailed understanding of the way in which a plasma mirror works could be deduced. It was found, that very subtle variations in the laser near field profile will have strong effects on the reflected pulse if the maximum fluence on the plasma mirror approaches the plasma ignition threshold. PMID- 21639487 TI - Compact fiber-coupled terahertz spectroscopy system pumped at 800 nm wavelength. AB - Photonic terahertz (THz) technology using femtosecond (fs) lasers has a great potential in a wide range of applications, such as non-destructive testing of objects or spectroscopic identification of chemical substances. For industrial purposes, a THz system has to be compact and easily implementable into the particular application. Therefore, fiber-coupled THz systems are the key to a widespread use of THz technology. In order to have flexible THz emitters and detectors near infrared fs light pulses have to be sent through optical fibers of considerable length. As a consequence, the fiber's dispersion has to be compensated for and nonlinear effects in the fiber have to be minimized. A fiber based THz time-domain spectroscopy system of high stability, flexibility, and portability is presented here. PMID- 21639488 TI - Measurement of the refractive index dispersion of As2Se3 bulk glass and thin films prior to and after laser irradiation and annealing using prism coupling in the near- and mid-infrared spectral range. AB - The prism coupling technique has been utilized to measure the refractive index in the near- and mid-IR spectral region of chalcogenide glasses in bulk and thin film form. A commercial system (Metricon model 2010) has been modified with additional laser sources, detectors, and a new GaP prism to allow the measurement of refractive index dispersion over the 1.5-10.6 MUm range. The instrumental error was found to be +/-0.001 refractive index units across the entire wavelength region examined. Measurements on thermally evaporated AMTIR2 thin films confirmed that (i) the film deposition process provides thin films with reduced index compared to that of the bulk glass used as a target, (ii) annealing of the films increases the refractive index of the film to the level of the bulk glass used as a target to create it, and (iii) it is possible to locally increase the refractive index of the chalcogenide glass using laser exposure at 632.8 nm. PMID- 21639489 TI - Measurement of optical-beat frequency in a photoconductive terahertz-wave generator using microwave higher harmonics. AB - A new method for measuring optical-beat frequencies in the terahertz (THz) region using microwave higher harmonics is presented. A microwave signal was applied to the antenna gap of a photoconductive (PC) device emitting a continuous electromagnetic wave at about 1 THz by the photomixing technique. The microwave higher harmonics with THz frequencies are generated in the PC device owing to the nonlinearity of the biased photoconductance, which is briefly described in this article. Thirteen nearly periodic peaks in the photocurrent were observed when the microwave was swept from 16 to 20 GHz at a power of -48 dBm. The nearly periodic peaks are generated by the homodyne detection of the optical beat with the microwave higher harmonics when the frequency of the harmonics coincides with the optical-beat frequency. Each peak frequency and its peak width were determined by fitting a Gaussian function, and the order of microwave harmonics was determined using a coarse (i.e., lower resolution) measurement of the optical beat frequency. By applying the Kalman algorithm to the peak frequencies of the higher harmonics and their standard deviations, the optical-beat frequency near 1 THz was estimated to be 1029.81 GHz with the standard deviation of 0.82 GHz. The proposed method is applicable to a conventional THz-wave generator with a photomixer. PMID- 21639490 TI - Continuous measurement of the arrival times of x-ray photon sequence. AB - In order to record x-ray pulse profile for x-ray pulsar-based navigation and timing, this paper presents a continuous, high-precision method for measuring arrival times of photon sequence with a common starting point. In this method, a high stability atomic clock is counted to measure the coarse time of arrival photon. A high resolution time-to-digital converter is used to measure the fine time of arrival photon. The coarse times and the fine times are recorded continuously and then transferred to computer memory by way of memory switch. The pulse profile is obtained by a special data processing method. A special circuit was developed and a low-level x-ray pulse profile measurement experiment system was setup. The arrival times of x-ray photon sequence can be consecutively recorded with a time resolution of 500 ps and the profile of x-ray pulse was constructed. The data also can be used for analysis by many other methods, such as statistical distribution of photon events per time interval, statistical distribution of time interval between two photon events, photon counting histogram, autocorrelation and higher order autocorrelation. PMID- 21639491 TI - The use of dynamic light scattering and brownian microscopy to characterize protein aggregation. AB - Dynamic light scattering (DLS) is often used to monitor aggregation in protein solutions. Here, we explore the veracity of the aggregate sizes, size distribution widths, concentrations, and lifetime resulting from DLS. We use as an example a solution of the protein lysozyme in which dense liquid clusters of radius about 100 nm reproducibly exist. We compare the results of DLS to those of brownian microscopy. We show that because of the sixth power dependence of the scattered light intensity on the size of the scatterers, DLS overestimates the mean size of the clusters. The factor of overestimation depends on the shape of the size distribution and is ~1.6 * in the studied solution. The related underestimate of the cluster concentration is ~10 *. The CONTIN algorithm, often employed to process DLS data, may, in some instances, produce non-physical results. We put forth an alternative method to determine the aggregates' sizes, concentrations, and volume fractions. We show that DLS yields a reliable width of the cluster size distribution only if the cluster concentration is above 10(9) cm(-3) and their volume fraction is above 10(-6). DLS yields a lower bound of the cluster lifetime, which may be orders of magnitude lower than the real one. PMID- 21639492 TI - Precision and accuracy in film stiffness measurement by Brillouin spectroscopy. AB - The interest in the measurement of the elastic properties of thin films is witnessed by a number of new techniques being proposed. However, the precision of results is seldom assessed in detail. Brillouin spectroscopy (BS) is an established optical, contactless, non-destructive technique, which provides a full elastic characterization of bulk materials and thin films. In the present work, the whole process of measurement of the elastic moduli by BS is critically analyzed: experimental setup, data recording, calibration, and calculation of the elastic moduli. It is shown that combining BS with ellipsometry a fully optical characterization can be obtained. The key factors affecting uncertainty of the results are identified and discussed. A procedure is proposed to discriminate factors affecting the precision from those affecting the accuracy. By the characterization of a model transparent material, silica in bulk and film form, it is demonstrated that both precision and accuracy of the elastic moduli measured by BS can reach 1% range, qualifying BS as a reference technique. PMID- 21639493 TI - A two-reflection divergent differentiating critical angle refractometer. AB - A new critical angle refractometer (CAR) for high accuracy refractive index measurement of liquid has been developed. The instrument improves the accuracy by two reflections in an elongated parallelogram prism, and acquires the angular reflectivity without any angle scanning parts through introduction of a point source with a divergent beam and a charge coupled device. In addition, it employs a simple and robust measurement method that gets the critical angle by differentiating the angular reflectivity. Through investigating absorbing media with absorption index kappa (the imaginary part of refractive index) from 0 to 10(-2.1), the theoretical calculation shows that the proposed two-reflection CAR would outperform the traditional one-reflection CAR on lowering the principal error from the differentiation method and improving the ability of getting the critical angle. By testing two typical liquids-salt-water solution and milk, the preliminary experiment indicates that this two-reflection divergent differentiating critical angle refractometer is feasible and of high accuracy. PMID- 21639494 TI - Performance of the Argonne National Laboratory electron cyclotron resonance charge breeder. AB - An electron cyclotron resonance charge breeder for the Californium rare ion breeder upgrade (CARIBU), a new radioactive beam facility for the Argonne Tandem Linac Accelerator System (ATLAS), has been constructed and commissioned. Charge breeding efficiencies up to 15.6% have been realized for stable beams with a typical breeding time of 10 ms/charge state. The CARIBU system has been undergoing commissioning tests utilizing a 100 mCi (252)Cf fission source. A charge breeding efficiency of 14.8 +/- 5% has been achieved for the first radioactive beam of (143)Cs(27+). PMID- 21639495 TI - Ion optical design of a collinear laser-negative ion beam apparatus. AB - An apparatus for photodetachment studies on atomic and molecular negative ions of medium up to heavy mass (M ? 500) has been designed and constructed. Laser and ion beams are merged in the apparatus in a collinear geometry and atoms, neutral molecules and negative ions are detected in the forward direction. The ion optical design and the components used to optimize the mass resolution and the transmission through the extended field-free interaction region are described. A 90 degrees sector field magnet with 50 cm bending radius in combination with two slits is used for mass dispersion providing a resolution of M/DeltaM?800 for molecular ions and M/DeltaM?400 for atomic ions. The difference in mass resolution for atomic and molecular ions is attributed to different energy distributions of the sputtered ions. With 1 mm slits, transmission from the source through the interaction region to the final ion detector was determined to be about 0.14%. PMID- 21639496 TI - Numerical studies of transient gain reduction process in a multi-wire proportional chamber. AB - A gain reduction process caused by successive beam irradiation in a multi-wire proportional chamber was numerically investigated to clarify the relations between the gas gain variation and the ion density distribution. A numerical code was developed based on a two-dimensional drift-diffusion model in order to evaluate the ion and electron density distributions and the electric field variation caused by the space charge effect. In order to consider the gain reduction process which occurs under the high rate and successive irradiation, the simulations were performed for the time period of ~10-100 MUs, which is much longer than the time required for ions to travel from an anode to a cathode. The numerical simulation results showed that for the low gas gain regime of ~10, quasi-stationary density distribution of the ions was formed by the high-rate beams of ~10(8)-10(10) particles per second, and that the transient variation of the gas gain became constant after establishment of the quasi-stationary ion density distributions. PMID- 21639497 TI - A compact electron cyclotron resonance proton source for the Paul Scherrer Institute's proton accelerator facility. AB - A compact electron cyclotron resonance proton source has been developed and installed recently at the Paul Scherrer Institute's high intensity proton accelerator. Operation at the ion source test stand and the accelerator demonstrates a high reliability and stability of the new source. When operated at a 10-12 mA net proton current the lifetime of the source exceeds 2000 h. The essential development steps towards the observed performance are described. PMID- 21639498 TI - Deconvolution of Thomson scattering temperature profiles. AB - Deconvolution of Thomson scattering (TS) profiles is required when the gradient length of the electron temperature (T(e)) or density (n(e)) are comparable to the instrument function length (Delta(R)). The most correct method for deconvolution to obtain underlying T(e) and n(e) profiles is by consideration of scattered signals. However, deconvolution at the scattered signal level is complex since it requires knowledge of all spectral and absolute calibration data. In this paper a simple technique is presented where only knowledge of the instrument function I(r) and the measured profiles, T(e, observed)(r) and n(e, observed)(r), are required to obtain underlying T(e)(r) and n(e)(r). This method is appropriate for most TS systems and is particularly important where high spatial sampling is obtained relative to Delta(R). PMID- 21639499 TI - Evaluation of an electrostatic dust removal system with potential application in next-step fusion devices. AB - The ability to manage inventories of carbon, tritium, and high-Z elements in fusion plasmas depends on means for effective dust removal. A dust conveyor, based on a moving electrostatic potential well, was tested with particles of tungsten, carbon, glass, and sand. A digital microscope imaged a representative portion of the conveyor, and dust particle size and volume distributions were derived before and after operation. About 10 mm(3) volume of carbon and tungsten particles were moved in under 5 s. The highest driving amplitude tested of 3 kV was the most effective. The optimal driving frequency was 210 Hz (maximum tested) for tungsten particles, decreasing to below 60 Hz for the larger sand particles. Measurements of particle size and volume distributions after 10 and 100 cycles show the breaking apart of agglomerated carbon and the change in particle distribution over short timescales (<1 s). PMID- 21639500 TI - Determination of radial location of rotating magnetic islands by use of poloidal soft x-ray detector arrays in the STOR-M tokamak. AB - A technique is presented for determining the radial location of the rotating magnetic islands in the STOR-M tokamak by use of soft x-ray (SXR) detector arrays. The location is determined by examining the difference in the integrated SXR emission intensities through two adjacent lines of sight. A model for calculating dependence of the line integrated SXR emission intensity on the radius, the mode numbers and the magnetic island geometry, has been developed. The SXR difference signal shows phase inversion when the impact parameter of the line of sight sweeps across the magnetic islands. Experimentally, the difference SXR signals significantly reduce noise and suppress the influence of background plasma fluctuations through common mode rejection when a dominant mode exists in the STOR-M tokamak. The radial locations of the m = 2 magnetic islands have been determined under several experimental conditions in the STOR-M discharges. With the decrease in the tokamak discharge current and thus the increase of the safety factor at the edge, the radial location of the m = 2 magnetic islands has been found to move radially inward. PMID- 21639501 TI - A piezo-thermal probe for thermomechanical analysis. AB - Thermomechanical analysis (TMA) is widely used to characterize materials and determine transition temperatures and thermal expansion coefficients. Atomic force microscopy (AFM) microcantilevers have been used for TMA. We have developed a micromachined probe that includes two embedded sensors: one for measuring the mechanical movement of the probe (deflection) and another for providing localized heating. The new probe reduces costs and complexity and allow for portability thereby eliminating the need for an AFM. The sensitivity of the deflection element ((DeltaR/R)/deflection) is 0.1 ppm/nm and its gauge factor is 3.24. The melting temperature of naphthalene is measured near 78.5 degrees C. PMID- 21639502 TI - The FAST module: an add-on unit for driving commercial scanning probe microscopes at video rate and beyond. AB - We present the design and the performance of the FAST (Fast Acquisition of SPM Timeseries) module, an add-on instrument that can drive commercial scanning probe microscopes (SPM) at and beyond video rate image frequencies. In the design of this module, we adopted and integrated several technical solutions previously proposed by different groups in order to overcome the problems encountered when driving SPMs at high scanning frequencies. The fast probe motion control and signal acquisition are implemented in a way that is totally transparent to the existing control electronics, allowing the user to switch immediately and seamlessly to the fast scanning mode when imaging in the conventional slow mode. The unit provides a completely non-invasive, fast scanning upgrade to common SPM instruments that are not specifically designed for high speed scanning. To test its performance, we used this module to drive a commercial scanning tunneling microscope (STM) system in a quasi-constant height mode to frame rates of 100 Hz and above, demonstrating extremely stable and high resolution imaging capabilities. The module is extremely versatile and its application is not limited to STM setups but can, in principle, be generalized to any scanning probe instrument. PMID- 21639503 TI - A modular designed ultra-high-vacuum spin-polarized scanning tunneling microscope with controllable magnetic fields for investigating epitaxial thin films. AB - A room-temperature ultra-high-vacuum scanning tunneling microscope for in situ scanning freshly grown epitaxial films has been developed. The core unit of the microscope, which consists of critical components including scanner and approach motors, is modular designed. This enables easy adaptation of the same microscope units to new growth systems with different sample-transfer geometries. Furthermore the core unit is designed to be fully compatible with cryogenic temperatures and high magnetic field operations. A double-stage spring suspension system with eddy current damping has been implemented to achieve <=5 pm z stability in a noisy environment and in the presence of an interconnected growth chamber. Both tips and samples can be quickly exchanged in situ; also a tunable external magnetic field can be introduced using a transferable permanent magnet shuttle. This allows spin-polarized tunneling with magnetically coated tips. The performance of this microscope is demonstrated by atomic-resolution imaging of surface reconstructions on wide band-gap GaN surfaces and spin-resolved experiments on antiferromagnetic Mn(3)N(2)(010) surfaces. PMID- 21639504 TI - Development of a mobile magnetic resonance imaging system for outdoor tree measurements. AB - By combining a 0.3 T permanent magnet with flexible rotation and translation mechanism, a probe with a local electromagnetic shielding, several electrical units, a mobile lift, and an electric wagon, a mobile magnetic resonance imaging (MRI) system was developed for outdoor tree measurements. 2D cross-sectional images of normal and diseased branches of a pear tree were acquired for measurements of T(1), T(2), proton density, and apparent diffusion constant (ADC). The ADC map clearly differentiated diseased from normal branches. A whole day measurement of the ADC map demonstrated that microscopic water flow in the normal branch changed proportionally with solar radiation. Therefore, we have concluded that our mobile MRI system is a powerful tool for studies of plants in outdoor environments. PMID- 21639505 TI - Atomic resolution ultrafast scanning tunneling microscope with scan rate breaking the resonant frequency of a quartz tuning fork resonator. AB - We present an ultra-fast scanning tunneling microscope with atomic resolution at 26 kHz scan rate which surpasses the resonant frequency of the quartz tuning fork resonator used as the fast scan actuator. The main improvements employed in achieving this new record are (1) fully low voltage design (2) independent scan control and data acquisition, where the tuning fork (carrying a tip) is blindly driven to scan by a function generator with the scan voltage and tunneling current (I(T)) being measured as image data (this is unlike the traditional point by-point move and measure method where data acquisition and scan control are switched many times). PMID- 21639506 TI - Three-view stereoscopy in dusty plasmas under microgravity: a calibration and reconstruction approach. AB - A three-camera stereoscopy setup is presented that allows to reconstruct the trajectories of particles in dusty plasmas under microgravity. The calibration procedure for the three-camera setup takes the special circumstances into account that occur in close-range imaging of small particles. Additionally, a reconstruction algorithm is presented that is based on the epipolar geometry and delivers the essential particle correspondences. Further improvements are achieved by analyzing the dynamic particle behavior. Two applications of our calibration and reconstruction procedure are presented: A two-dimensional dust structure in the laboratory with a large percentage of hidden particles, and particles inside the void of a dust cloud under microgravity. PMID- 21639507 TI - Errors in particle tracking velocimetry with high-speed cameras. AB - Velocity errors in particle tracking velocimetry (PTV) are studied. When using high-speed video cameras, the velocity error may increase at a high camera frame rate. This increase in velocity error is due to particle-position uncertainty, which is one of the two sources of velocity errors studied here. The other source of error is particle acceleration, which has the opposite trend of diminishing at higher frame rates. Both kinds of errors can propagate into quantities calculated from velocity, such as the kinetic temperature of particles or correlation functions. As demonstrated in a dusty plasma experiment, the kinetic temperature of particles has no unique value when measured using PTV, but depends on the sampling time interval or frame rate. It is also shown that an artifact appears in an autocorrelation function computed from particle positions and velocities, and it becomes more severe when a small sampling-time interval is used. Schemes to reduce these errors are demonstrated. PMID- 21639508 TI - Low axial drift stage and temperature controlled liquid cell for z-scan fluorescence correlation spectroscopy in an inverted confocal geometry. AB - A recent iteration of fluorescence correlation spectroscopy (FCS), z-scan FCS, has drawn attention for its elegant solution to the problem of quantitative sample positioning when investigating two-dimensional systems while simultaneously providing an excellent method for extracting calibration-free diffusion coefficients. Unfortunately, the measurement of planar systems using (FCS and) z-scan FCS still requires extremely mechanically stable sample positioning, relative to a microscope objective. As axial sample position serves as the inherent length calibration, instabilities in sample position will affect measured diffusion coefficients. Here, we detail the design and function of a highly stable and mechanically simple inverted microscope stage that includes a temperature controlled liquid cell. The stage and sample cell are ideally suited to planar membrane investigations, but generally amenable to any quantitative microscopy that requires low drift and excellent axial and lateral stability. In the present work we evaluate the performance of our custom stage system and compare it with the stock microscope stage and typical sample sealing and holding methods. PMID- 21639509 TI - Reconstruction of a scanned topographic image distorted by the creep effect of a Z scanner in atomic force microscopy. AB - We analyzed the illusory slopes of scanned images caused by the creep of a Z scanner in an atomic force microscope (AFM) operated in constant-force mode. A method to reconstruct a real topographic image using two scanned images was also developed. In atomic force microscopy, scanned images are distorted by undesirable effects such as creep, hysteresis of the Z scanner, and sample tilt. In contrast to other undesirable effects, the illusory slope that appears in the slow scanning direction of an AFM scan is highly related to the creep effect of the Z scanner. In the controller for a Z scanner, a position-sensitive detector is utilized to maintain a user-defined set-point or force between a tip and a sample surface. This serves to eliminate undesirable effects. The position sensitive detector that detects the deflection of the cantilever is used to precisely measure the topography of a sample. In the conventional constant-force mode of an atomic force microscope, the amplitude of a control signal is used to construct a scanned image. However, the control signal contains not only the topography data of the sample, but also undesirable effects. Consequently, the scanned image includes the illusory slope due to the creep effect of the Z scanner. In an automatic scanning process, which requires fast scanning and high repeatability, an atomic force microscope must scan the sample surface immediately after a fast approach operation has been completed. As such, the scanned image is badly distorted by a rapid change in the early stages of the creep effect. In this paper, a new method to obtain the tilt angle of a sample and the creep factor of the Z scanner using only two scanned images with no special tools is proposed. The two scanned images can be obtained by scanning the same area of a sample in two different slow scanning directions. We can then reconstruct a real topographic image based on the scanned image, in which both the creep effect of the Z scanner and the slope effect of the sample have been eliminated. The slope effect of the sample should be eliminated so as to avoid further distortion after removal of the creep effect. The creep effect can be removed from the scanned image using the proposed method, and a real topographic image can subsequently be efficiently reconstructed. PMID- 21639510 TI - Antenna-based ultrahigh vacuum microwave frequency scanning tunneling microscopy system. AB - The instrumental synthesis of high resolution scanning tunneling microscopy (STM) with the ability to measure differential capacitance with atomic scale resolution is highly desirable for fundamental metrology and for the study of novel physical characteristics. Microwave frequency radiation directed at the tip-sample junction in an STM system allows for such high-resolution differential capacitance information. This ability is particularly critical in ultrahigh vacuum environments, where the additional parameter space afforded by including a capacitance measurement would prove powerful. Here we describe the modifications made to a commercial scanning tunneling microscope to allow for broad microwave frequency alternating current scanning tunneling microscopy (ACSTM) in ultrahigh vacuum conditions using a relatively simple loop antenna and microwave difference frequency detection. The advantages of our system are twofold. First, the use of a removable antenna on a commercial STM prevents interference with other UHV processes while providing a simple method to retrofit any commercial UHV-STM with UHV-ACSTM capability. Second, mounting the microwave antenna on a translator allows for specific tuning of the system to replicate experimental conditions between samples, which is particularly critical in sensitive systems like organic thin films or single molecules where small changes in incident power can affect the results. Our innovation therefore provides a valuable approach to give nearly any commercial STM, be it an ambient or UHV system, the capability to measure atomic-scale microwave studies such as differential capacitance or even single molecule microwave response, and it ensures that experimental ACSTM conditions can be held constant between different samples. PMID- 21639511 TI - Simultaneous measurement of normal and friction forces using a cantilever-based optical interfacial force microscope. AB - We measured normal and friction forces simultaneously using a recently developed cantilever-based optical interfacial force microscope technique for studies of interfacial structures and mechanical properties of nanoscale materials. We derived how the forces can be incorporated into the detection signal using the classical Euler equation for beams. A lateral modulation with the amplitude of nanometers was applied to create the friction forces between tip and sample. We demonstrated its capability by measuring normal and friction forces of interfacial water at the molecular scale over all distance ranges. PMID- 21639512 TI - A dynamic punch method to quantify the dynamic shear strength of brittle solids. AB - Shear strength is an important material parameter for brittle solids. This parameter has been extensively used in material failure models. Although a few methods have been proposed to quantify this parameter under the static loading condition, there is no such a method available to measure it under dynamic loading conditions. This paper presents a punch shear device to measure the dynamic shear strength of brittle solids. In this method, a split Hopkinson pressure bar system (SHPB) is used to exert the dynamic load to a thin disc sample, which is placed in a specially designed holder to minimize the bending stress induced by punching. The sample holder also allows the punch head to load the sample directly and in combination with momentum-trap technique in SHPB, it enables soft recovery of the rock plug and rock ring produced by the punching test. The flexibility and applicability of this method is demonstrated by the application of an isotropic and fine-grained sandstone. Within the theoretical framework of the classical Mohr-Coulomb failure model, the obtained dynamic shear strengths are consistent with the dynamic tensile strengths for the same rock from the literature. PMID- 21639513 TI - Integrated electronic transport and thermometry at milliKelvin temperatures and in strong magnetic fields. AB - We fabricated a He-3 immersion cell for transport measurements of semiconductor nanostructures at ultra low temperatures and in strong magnetic fields. We have a new scheme of field-independent thermometry based on quartz tuning fork Helium-3 viscometry which monitors the local temperature of the sample's environment in real time. The operation and measurement circuitry of the quartz viscometer is described in detail. We provide evidence that the temperature of two-dimensional electron gas confined to a GaAs quantum well follows the temperature of the quartz viscometer down to 4 mK. PMID- 21639514 TI - Image time-correlation, dynamic light scattering, and birefringence for the study of the response of anisometric colloids to external fields. AB - In this paper, a detailed description of equipment is given, specially designed to characterize the response of non-spherical colloids to external fields. To characterize slow structural changes on a large length scale we developed an image correlation method, fast dynamics on the colloidal-particle level is probed by means of a vertically mounted, small angle dynamic light scattering setup, while the orientational order (induced by the external field) is measured with a birefringence setup with off-normal incidence. The performance of this in situ set of instruments is illustrated by experiments on concentrated dispersions of very long and thin, charged colloidal rods (fd-virus particles) in external electric fields. Here, the frequency of the field is sufficiently low to polarize electrical double layers, leading to additional inter-colloidal interactions which are found to give rise to phase/state transitions and dynamical states (K. Kang and J. K. G. Dhont, Soft Matter 6, 273, 2010). PMID- 21639515 TI - An integrated capacitance bridge for high-resolution, wide temperature range quantum capacitance measurements. AB - We have developed a highly sensitive integrated capacitance bridge for quantum capacitance measurements. Our bridge, based on a GaAs HEMT amplifier, delivers attofarad (aF) resolution using a small AC excitation at or below k(B)T over a broad temperature range (4-300 K). We have achieved a resolution at room temperature of 60 aF/?Hz for a 10 mV ac excitation at 17.5 kHz, with an improved resolution at cryogenic temperatures, for the same excitation amplitude. We demonstrate the utility of our bridge for measuring the quantum capacitance of nanostructures by measuring the capacitance of top-gated graphene devices and cleanly resolving the density of states. PMID- 21639516 TI - Micromirror arrays to assess luminescent nano-objects. AB - We propose an array of submicrometer mirrors to assess luminescent nano-objects. Micromirror arrays (MMAs) are fabricated on Si (001) wafers via selectively doping Ga using the focused ion beam technique to form p-type etch stop regions, subsequent anisotropic chemical etching, and Al deposition. MMAs provide two benefits: reflection of luminescence from nano-objects within MMAs toward the Si (001) surface normal and nano-object labeling. The former increases the probability of optics collecting luminescence and is demonstrated by simulations based on the ray-tracing and finite-difference time-domain methods as well as by experiments. The latter enables different measurements to be repeatedly performed on a single nano-object located at a certain micromirror. For example, a single InGaN/GaN nanocolumn is assessed by scanning electron microscopy and microphotoluminescence spectroscopy. PMID- 21639517 TI - Miniature ceramic-anvil high-pressure cell for magnetic measurements in a commercial superconducting quantum interference device magnetometer. AB - A miniature opposed-anvil high-pressure cell has been developed for magnetic measurement in a commercial superconducting quantum interference device magnetometer. Non-magnetic anvils made of composite ceramic material were used to generate high-pressure with a Cu-Be gasket. We have examined anvils with different culet sizes (1.8, 1.6, 1.4, 1.2, 1.0, 0.8, and 0.6 mm). The pressure generated at low temperature was determined by the pressure dependence of the superconducting transition of lead (Pb). The maximum pressure P(max) depends on the culet size of the anvil: the values of P(max) are 2.4 and 7.6 GPa for 1.8 and 0.6 mm culet anvils, respectively. We revealed that the composite ceramic anvil has potential to generate high-pressure above 5 GPa. The background magnetization of the Cu-Be gasket is generally two orders of magnitude smaller than the Ni-Cr Al gasket for the indenter cell. The present cell can be used not only with ferromagnetic and superconducting materials with large magnetization but also with antiferromagnetic compounds with smaller magnetization. The production cost of the present pressure cell is about one tenth of that of a diamond anvil cell. The anvil alignment mechanism is not necessary in the present pressure cell because of the strong fracture toughness (6.5 MPa m(1/2)) of the composite ceramic anvil. The simplified pressure cell is easy-to-use for researchers who are not familiar with high-pressure technology. Representative results on the magnetization of superconducting MgB(2) and antiferromagnet CePd(5)Al(2) are reported. PMID- 21639518 TI - A versatile low-temperature setup for the electrical characterization of single molecule junctions. AB - We present a modular high-vacuum setup for the electrical characterization of single molecules down to liquid helium temperatures. The experimental design is based on microfabricated mechanically controllable break junctions, which offer control over the distance of two electrodes via the bending of a flexible substrate. The actuator part of the setup is divided into two stages. The slow stage is based on a differential screw drive with a large bending range. An amplified piezoceramic actuator forms the fast stage of the setup, which can operate at bending speeds of up to 800 MUm/s. In our microfabricated break junctions this is translated into breaking speeds of several 10 nm/s, sufficient for the fast acquisition of large statistical datasets. The bandwidth of the measurement electronics has been optimized to enable fast dI/dV spectroscopy on molecular junctions with resistances up to 100 MOmega. The performance of the setup is demonstrated for a pi-conjugated oligo(phenylene-ethynylene)-dithiol molecule. PMID- 21639519 TI - The NanoBeamBalance: a passive, tensile-test device for the atomic force microscope. AB - An add-on device is presented, which significantly expands the force measurement capabilities of the atomic force microscope (AFM). The device consists of a completely passive mechanism, which translates the vertical motion of the AFM tip in force measurements into a horizontal motion of two sample support pads. The advantage is that it is much easier to deposit microscopic samples from suspension onto flat surfaces than to attach them reliably between tip and a surface. The working-principle and the design of the device is comprehensively described and demonstrated on the example of collagen fibres with a diameter of a few MUm. Well-defined tensile measurements in longitudinal direction were performed, showing that the tensile stiffness of collagen fibres from rat tail tendon decreases by a factor of 5 when rehydrated from a dried sample and slowly increases upon cross-linking with glutaraldehyde. PMID- 21639520 TI - High sensitivity magnetometer for measuring the isotropic and anisotropic magnetisation of small samples. AB - We describe how the full, isotropic and anisotropic, magnetisation of samples as small as tens of micrometers in size can be sensitively measured using a piezoresistive microcantilever and a small, moveable ferromagnet. Depending on the position of the ferromagnet, a strong but highly local field gradient of up to ~4200 T/m can be applied at the sample or removed completely during a single measurement. In this way, the magnetic force and torque on the sample can be independently determined without moving the sample or cycling the experimental system. The technique can be used from millikelvin temperatures to ~85 K and in magnetic fields from 2 T to the highest fields available. We demonstrate its application in measurements of the semimagnetic semiconductor Hg(1 - x)Fe(x)Se, where we achieved a moment sensitivity of better than 2.5 * 10(-14) J/T for both isotropic and anisotropic components. PMID- 21639521 TI - Hybrid quadrupole mass filter/quadrupole ion trap/time-of-flight-mass spectrometer for infrared multiple photon dissociation spectroscopy of mass selected ions. AB - We present a laboratory-constructed mass spectrometer optimized for recording infrared multiple photon dissociation (IRMPD) spectra of mass-selected ions using a benchtop tunable infrared optical parametric oscillator/amplifier (OPO/A). The instrument is equipped with two ionization sources, an electrospray ionization source, as well as an electron ionization source for troubleshooting. This hybrid mass spectrometer is composed of a quadrupole mass filter for mass selection, a reduced pressure (~10(-5) Torr) quadrupole ion trap (QIT) for OPO irradiation, and a reflectron time-of-flight drift tube for detecting the remaining precursor and photofragment ions. A helium gas pulse is introduced into the QIT to temporarily increase the pressure and hence enhance the trapping efficiency of axially injected ions. After a brief pump-down delay, the compact ion cloud is subjected to the focused output from the continuous wave OPO. In a recent study, we implemented this setup in the study of protonated tryptophan, TrpH(+), as well as collision-induced dissociation products of this protonated amino acid [W. K. Mino, Jr., K. Gulyuz, D. Wang, C. N. Stedwell, and N. C. Polfer, J. Phys. Chem. Lett. 2, 299 (2011)]. Here, we give a more detailed account on the figures of merit of such IRMPD experiments. The appreciable photodissociation yields in these measurements demonstrate that IRMPD spectroscopy of covalently bound ions can be routinely carried out using benchtop OPO setups. PMID- 21639522 TI - Vacuum compatible sample positioning device for matrix assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry imaging. AB - The high mass accuracy and resolving power of Fourier transform ion cyclotron resonance mass spectrometers (FT-ICR MS) make them ideal mass detectors for mass spectrometry imaging (MSI), promising to provide unmatched molecular resolution capabilities. The intrinsic low tolerance of FT-ICR MS to RF interference, however, along with typically vertical positioning of the sample, and MSI acquisition speed requirements present numerous engineering challenges in creating robotics capable of achieving the spatial resolution to match. This work discusses a two-dimensional positioning stage designed to address these issues. The stage is capable of operating in ~1 * 10(-8) mbar vacuum. The range of motion is set to 100 mm * 100 mm to accommodate large samples, while the positioning accuracy is demonstrated to be less than 0.4 micron in both directions under vertical load over the entire range. This device was integrated into three different matrix assisted laser desorption/ionization (MALDI) FT-ICR instruments and showed no detectable RF noise. The "oversampling" MALDI-MSI experiments, under which the sample is completely ablated at each position, followed by the target movement of the distance smaller than the laser beam, conducted on the custom-built 7T FT-ICR MS demonstrate the stability and positional accuracy of the stage robotics which delivers high spatial resolution mass spectral images at a fraction of the laser spot diameter. PMID- 21639523 TI - A versatile pulse programmer for magnetic resonance imaging. AB - Hardware and software solutions for a versatile pulse programmer have been presented. The core of the pulse programmer is an FPGA device that provides flexibility to the design and reduces the number of electronics elements needed. The event of the pulse programmer consists of 16 bits. The main feature of the proposed pulse programmer is that the 16 outputs can be independently delayed. This is important for correcting delays of the RF channels or the gradient channels due to various causes. The proposed pulse programmer is integrated into an MRI scanner, and the correction of the gradient system delay is taken as an example to experimentally demonstrate its performance. PMID- 21639524 TI - Measurement of circular dichroism dynamics in a nanosecond temperature-jump experiment. AB - The use of a fast temperature jump (T-jump) is a very powerful experiment aiming at studying protein denaturation dynamics. However, probing the secondary structure is a difficult challenge and rarely yields quantitative values. We present the technical implementation of far-UV circular dichroism in a nanosecond T-jump experiment and show that this experiment allows us to follow quantitatively the change in the helical fraction of a poly(glutamic acid) peptide during its thermal denaturation with 12 ns time resolution. PMID- 21639525 TI - Calibration system with cryogenically-cooled loads for cosmic microwave background polarization detectors. AB - We present a novel system to calibrate millimeter-wave polarimeters for cosmic microwave background (CMB) polarization measurements. This technique is an extension of the conventional metal mirror rotation approach, however, it employs cryogenically-cooled blackbody absorbers. The primary advantage of this system is that it can generate a slightly polarized signal (~100 mK) in the laboratory; this is at a similar level to that measured by ground-based CMB polarization experiments observing a ~10 K sky. It is important to reproduce the observing condition in the laboratory for reliable characterization of polarimeters before deployment. In this paper, we present the design and principle of the system and demonstrate its use with a coherent-type polarimeter used for an actual CMB polarization experiment. This technique can also be applied to incoherent-type polarimeters and it is very promising for the next-generation CMB polarization experiments. PMID- 21639526 TI - The depolarization of Pb(Zr0.52Ti0.48)O3 ferroelectrics by cylindrical radially expanding shock waves and its utilization for miniature pulsed power. AB - The effects of depolarization of Pb(Zr(0.52)Ti(0.48))O(3) (PZT 52/48) poled ferroelectrics by cylindrical radially expanding shock waves propagated along and across the polarization vector P(0) were experimentally detected. Miniature (total volume 100 cm(3)) autonomous generators based on these effects were capable of producing output voltage pulses with amplitudes up to 25 kV and output energies exceeding 1 J. PMID- 21639527 TI - The flexible Ca-test: an improved performance in a gas permeability measurement system. AB - A flexible performance permeability measuring test for flexible organic light emitting diodes is described in this paper. A single thin film layer of gas barriers is constructed on polyethersulfone (PES). The barrier coats the upper and lower surfaces of the PES layer. Two PES samples, one coated with Al(2)O(3) on both surfaces and the other coated on a single surface, were made for comparison. According to this test, the time-dependent transmission curve of the one sided barrier sample had a linear slope which measured 1.65 g/m(2)/day at room temperature at a 50% relative humidity. This result shows that the measurement time is about 182% faster than has been achieved with the conventional test structure that uses a glass substrate. In addition, this measurement structure not only reduces the inevitable electrical noise which occurs during measurement but also increases the water vapor permeation signal. These effects improve the sensing reliability of the test. In addition, this structure is flexible, so one can instantly detect barrier performance changes when applying external stress. PMID- 21639528 TI - Simultaneous measurement of temperature and emissivity of lunar regolith simulant using dual-channel millimeter-wave radiometry. AB - Millimeter wave (MMW) radiometry can be used for simultaneous measurement of emissivity and temperature of materials under extreme environments (high temperature, pressure, and corrosive environments). The state-of-the-art dual channel MMW passive radiometer with active interferometric capabilities at 137 GHz described here allows for radiometric measurements of sample temperature and emissivity up to at least 1600 degrees C with simultaneous measurement of sample surface dynamics. These capabilities have been used to demonstrate dynamic measurement of melting of powders of simulated lunar regolith and static measurement of emissivity of solid samples. The paper presents the theoretical background and basis for the dual-receiver system, describes the hardware in detail, and demonstrates the data analysis. Post-experiment analysis of emissivity versus temperature allows further extraction from the radiometric data of millimeter wave viewing beam coupling factors, which provide corroboratory evidence to the interferometric data of the process dynamics observed. These results show the promise of the MMW system for extracting quantitative and qualitative process parameters for industrial processes and access to real-time dynamics of materials behavior in extreme environments. PMID- 21639529 TI - Ferromagnetic resonance of a single magnetic nanowire measured with an on-chip microwave interferometer. AB - An on-chip microwave interferometer suitable for high-sensitivity nanoscale magnetic material characterization is proposed. The device cancels the background parasitic common mode noise automatically. The magnetization dynamics of a 240 nm wide, 5 MUm long, and 70 nm thick single permalloy nanowire is investigated. Compared with a prototype device proposed previously, the proposed device has a more than 20 dB sensitivity improvement. Full wave three-dimensional simulation shows that the device has the capability of studying the fundamental physics of nanoscale magnetic devices. PMID- 21639530 TI - High speed heterodyne infrared thermography applied to thermal diffusivity identification. AB - We have combined InfraRed thermography and thermal wave techniques to perform microscale, ultrafast (microsecond) temperature field measurements. The method is based on an IR camera coupled to a microscope and synchronized to the heat source by means of phase locked function generators. The principle is based on electronic stroboscopic sampling where the low IR camera acquisition frequency f(acq) (25 Hz) undersamples a high frequency thermal wave. This technique permits the measurement of the emissive thermal response at a (microsecond) short time scale (microsecond) with the full frame mode of the IR camera with a spatial thermal resolution of 7 MUm. Then it becomes possible to study 3D transient heat transfer in heterogeneous and high thermal conductive thin layers. Thus it is possible for the first time in our knowledge to achieve temperature field measurements in heterogeneous media within a wide range of time domains. The IR camera is now a suitable instrument for multiscale thermal analysis. PMID- 21639531 TI - Coded excitation for infrared non-destructive testing of carbon fiber reinforced plastics. AB - This paper proposes a Barker coded excitation for defect detection using infrared non-destructive testing. Capability of the proposed excitation scheme is highlighted with recently introduced correlation based post processing approach and compared with the existing phase based analysis by taking the signal to noise ratio into consideration. Applicability of the proposed scheme has been experimentally validated on a carbon fiber reinforced plastic specimen containing flat bottom holes located at different depths. PMID- 21639532 TI - BioRef: a versatile time-of-flight reflectometer for soft matter applications at Helmholtz-Zentrum Berlin. AB - BioRef is a versatile novel time-of-flight reflectometer featuring a sample environment for in situ infrared spectroscopy at the reactor neutron source BER II of the Helmholtz Zentrum Berlin fur Materialien und Energie (HZB). After two years of design and construction phase the instrument has recently undergone commissioning and is now available for specular and off-specular neutron reflectivity measurements. BioRef is especially dedicated to the investigation of soft matter systems and studies at the solid-liquid interface. Due to flexible resolution modes and variable addressable wavelength bands that allow for focusing onto a selected scattering vector range, BioRef enables a broad range of surface and interface investigations and even kinetic studies with subsecond time resolution. The instrumental settings can be tailored to the specific requirements of a wide range of applications. The performance is demonstrated by several reference measurements, and the unique option of in situ on-board infrared spectroscopy is illustrated by the example of a phase transition study in a lipid multilayer film. PMID- 21639533 TI - Compact and accurate concept of laser wavemeters based on ellipsometry. AB - Common laser wavemeters are based on a scanning Michelson interferometer. Displacements of the moving mirror as long as tens of centimeters are needed to reach a relative accuracy of 1 * 10(-6) (1sigma) on the unknown laser wavelengths. Such a long displacement range makes the system very sensitive to mechanical vibrations and to misalignments of the laser beams. The purpose of this paper is to demonstrate a new concept of laser wavemeter based on the measurements of the ellipsometric parameters psi and Delta of the laser beams. Experimental results show that a 10(-6) (1sigma) accuracy level could be reach with a displacement range of only 4 MUm. Implementations of the device are described. Comparisons between our polarimetric wavemeter and a calibrated wavemeter are presented for two lasers, an extended cavity laser diode at 656 nm and a 532 nm green line Nd:YAG laser. PMID- 21639534 TI - Deconvolution of damping forces with a nonlinear microresonator. AB - We report a fully electrical microcantilever device that utilizes capacitance for both actuation and detection and show that it can characterize various gases with a bare silicon microcantilever. We find the motion of the cantilever as it rings down when the oscillating force is removed, by measuring the voltage induced by the oscillating capacitance in the microcantilever/counterelectrode system. The ringdown waveform was analyzed using an iterative numerical algorithm to calculate the oscillator motion, modeling the cantilever/electrode capacitance to calculate the electrostatic force. We find that nonlinearity in the motion of the cantilever is not necessarily a disadvantage. After calibration, we simultaneously measure viscosity and density of several gaseous mixtures, yielding viscosities within +/-2% and densities within +/-6% of NIST values. PMID- 21639535 TI - Two-dimensional fringe probing of transient liquid temperatures in a mini space. AB - A 2D fringe probing transient temperature measurement technique based on photothermal deflection theory was developed. It utilizes material's refractive index dependence on temperature gradient to obtain temperature information from laser deflection. Instead of single beam, this method applies multiple laser beams to obtain 2D temperature information. The laser fringe was generated with a Mach-Zehnder interferometer. A transient heating experiment was conducted using an electric wire to demonstrate this technique. Temperature field around a heating wire and variation with time was obtained utilizing the scattering fringe patterns. This technique provides non-invasive 2D temperature measurements with spatial and temporal resolutions of 3.5 MUm and 4 ms, respectively. It is possible to achieve temporal resolution to 500 MUs utilizing the existing high speed camera. PMID- 21639536 TI - Characteristics of an actuator-driven pulsed water jet generator to dissecting soft tissue. AB - This paper reports characteristics of an actuator-driven pulsed water jet generator applied, in particular, to dissect soft tissues. Results of experiments, by making use of high speed recording of optical visualization and varying nozzle diameter, actuator time interval, and their effects on dissection performance are presented. Jet penetration characteristics are compared with continuous water jet and hence potential assessment of pulsed water jets to clinical applications is performed. PMID- 21639537 TI - Cryogenic apparatus for study of near-field heat transfer. AB - For bodies spaced in vacuum at distances shorter than the wavelength of the thermal radiation, radiative heat transfer substantially increases due to the contribution of evanescent electromagnetic waves. Experimental data on heat transfer in near-field regime are scarce. We have designed a cryogenic apparatus for the study of heat transfer over microscopic distances between metallic and non-metallic surfaces. Using a mechanical positioning system, a planeparallel gap between the samples, concentric disks, each 35 mm in diameter, is set and varied from 10(0) to 10(3) MUm. The heat transferred from the hot (10 - 100 K) to the cold sample (~5 K) sinks into a liquid helium bath through a thermal resistor, serving as a heat flux meter. Transferred heat power within ~2 nW/cm(2) and ~30 MUW/cm(2) is derived from the temperature drop along the thermal resistor. For tungsten samples, the distance of the near-field effect onset was inversely proportional to temperature and the heat power increase was observed up to three orders of magnitude greater than the power of far-field radiative heat transfer. PMID- 21639538 TI - Accurate surface tension measurement of glass melts by the pendant drop method. AB - A pendant drop tensiometer, coupled with image digitization technology and a best fitting algorithm, was built to accurately measure the surface tension of glass melts at high temperatures. More than one thousand edge-coordinate points were obtained for a pendant glass drop. These edge points were fitted with the theoretical drop profiles derived from the Young-Laplace equation to determine the surface tension of glass melt. The uncertainty of the surface tension measurements was investigated. The measurement uncertainty (sigma) could be related to a newly defined factor of drop profile completeness (Fc): the larger the Fc is, the smaller sigma is. Experimental data showed that the uncertainty of the surface tension measurement when using this pendant drop tensiometer could be +/-3 mN/m for glass melts. PMID- 21639539 TI - Zero-power autonomous buoyancy system controlled by microbial gas production. AB - A zero-power ballast control system that could be used to float and submerge a device solely using a gas source was built and tested. This system could be used to convey sensors, data loggers, and communication devices necessary for water quality monitoring and other applications by periodically maneuvering up and down a water column. Operational parameters for the system such as duration of the submerged and buoyant states can be varied according to its design. The gas source can be of any origin, e.g., compressed air, underwater gas vent, gas produced by microbes, etc. The zero-power ballast system was initially tested using a gas pump and further tested using gas produced by Clostridium acetobutylicum. Using microbial gas production as the only source of gas and no electrical power during operation, the system successfully floated and submerged periodically with a period of 30 min for at least 24 h. Together with microbial fuel cells, this system opens up possibilities for underwater monitoring systems that could function indefinitely. PMID- 21639540 TI - Experimental and numerical analysis of the temperature distribution of injection molded products using protruding microprobes. AB - Injection molding has been one of the most important polymer processing methods for manufacturing plastic parts. In the process, the temperature is an important parameter that influences process features such as cycle times, crystallization rates, degree of crystallinity, melt flow properties, and molded product qualities. This study aims to, experimentally and numerically, examine the three dimensional temperature distribution along the melt flow path of injection molded parts. A special experimental set-up, which includes an injection mold equipped with protruding microprobes for guiding embedded thermocouples, was designed and built to measure the temperature field along the flow path, i.e., inside the runner and the cavity, of injection molded products. The experimental results suggested that the disturbance induced by the probes remained negligible and precise temperature profiles could be measured at various positions inside the cavity. A significant increase of melt temperature was found to result from the viscous dissipation of the polymeric materials in the runner. Additionally, a commercially available code was employed to simulate and predict the temperature variation in injection molded parts. It was shown that the numerical simulation predicted better the temperature distributions inside the cavity than those along the runner. PMID- 21639541 TI - Broad-band and high-temperature ultrasonic transducer fabricated using a Pb(In(1/2)Nb(1/2))-Pb(Mg(1/3)Nb(2/3))-PbTiO3 single crystal/epoxy 1-3 composite. AB - In this paper, 1-3 composites based on Pb(In(1/2)Nb(1/2))-Pb(Mg(1/3)Nb(2/3)) PbTiO(3) (PIMNT) single crystal and high-temperature epoxy were fabricated with various volume fractions of PIMNT single crystal ranging from 0.4 to 0.9. The electrical properties were studied as functions of PIMNT volume fraction and temperature, and it revealed that the nature of ultrahigh electromechanical coupling coefficient (0.82-0.93) and low acoustic impedance (17-19 MRayl) of the composites can be retained within a wide temperature range from room temperature to 185 degrees C. Single element ultrasonic transducer using the PIMNT 1-3 composite was fabricated and characterized as a function of temperature. It was found that the transducer can still work normally at high temperatures, such as 165 degrees C, possessing a bandwidth of 95% and insertion loss of -27 dB. PMID- 21639542 TI - Comparison between beryllium and diamond-backing plates in diamond-anvil cells: application to single-crystal x-ray diffraction high-pressure data. AB - A direct comparison between two complete intensity datasets, collected on the same sample loaded in two identical diamond-anvil pressure cells equipped, respectively, with beryllium and diamond-backing plates was performed. The results clearly demonstrate that the use of diamond-backing plates significantly improves the quality of crystal structure data. There is a decrease in the internal R factor for averaging, structure refinement agreement factors, and in the errors and uncertainties of the atomic coordinates, atomic displacement parameters, and individual bond lengths. PMID- 21639543 TI - Collection method for extra aligned nanofibers deposited by electrospinning. AB - Electrospinning is a very promising nanotechnology method, mostly thanks to industrial scale up ability, relatively low purchase cost, and huge variability. Nevertheless, its principal disadvantage is the way in which the individual nanofibers are randomly deposited while being collected onto a collector after passing through the chaotic whipping phase. Present and future applications of nanomaterials will need their precise, ordered intrinsic structures, or may even require accurately determined anisotropic properties because of significant improvement of material properties such as mechanical, electronic, optical, etc. This work demonstrates a novel and advanced collector including a sliding board mechanism. The resulting nanofibrous materials obtain extremely high orientation order parameter (S > 97%). Other important advantages of this technological method are discussed here in more detail. PMID- 21639544 TI - Nuclear quadrupole double resonance spectrometer with magnetic property measurement system direct current superconducting quantum interference device detector and automatic tuning. AB - A new nuclear quadrupole double resonance spectrometer based on a commercial superconducting quantum interference device (SQUID) setup (a Magnetic Property Measurement System (MPMS) from Quantum Design) is described. The experiment involves the indirect detection of pure quadrupole resonances (PQR) of a dilute spin system via the direct SQUID detected NMR of an abundant spin system. The experiment is conducted at low (3-20 K) temperatures and the magnetic field is cycled between a high (5.5 T) polarizing field, to an intermediate (0.1 T) detection field, to zero field where the sample is irradiated with a modulated search RF and back to the detection field. Loss of the NMR signal indicates the detection of a PQR. The RF circuit used for both the NMR and zero field irradiation is digitally controlled. Use of the External Device Control allows for the complete automation of the system. Test measurements on diphenyl ether are in good agreement with previously reported results. Pure (17)O quadrupole resonances were detected for spin systems with concentrations as low as 120 MUM. PMID- 21639545 TI - Design and construct of a new detector for gas chromatography based on continuous negative corona discharge. AB - In this work, a new detector was designed and constructed based on negative corona discharge. This detector can be used separately or as a detector in gas chromatography. The detector and chromatographic variables including cell temperature, gas flow rates, voltage between the two electrodes, and column temperature were optimized. Chloroform was used as a test compound to evaluate the performance of the detector. The detection limit of chloroform was obtained 0.78 ng/ml and its dynamic range was over the range of 2-840 ng/ml. The relative standard detection was about 6% for the limit of quantification. This detector is able to be used as an alternative for analysis of compounds containing electronegative elements. PMID- 21639546 TI - Tomographic reconstruction of incompressible flow. AB - In many experiments facilitating tomography the reconstruction problem is under determined, meaning there are many possible solutions consistent with the measurements. If the sampling rate is fast relative to the typical evolution time, the known physical dynamics of the system can be used as additional reconstruction constraints. Here we demonstrate that incorporating the requirement of incompressible flow can improve significantly the fidelity of the reconstructed sequence. The incompressibility of the reconstruction is assured by requiring the conservation of the density moments. It is demonstrated that the "incompressible" reconstruction can be significantly more accurate than the reconstruction using standard methods. A consequence of the density moments' conservation is the conservation of the density histogram throughout the reconstructed sequence. PMID- 21639547 TI - Characteristics of a symmetrical Cockcroft-Walton power supply of 50 Hz 1.2 MV/50 mA. AB - A 1.2 MV/50 mA symmetrical Cockcroft-Walton (SCW) power supply of over 83% power efficiency, driven by 50 Hz frequency, was developed for an industrial electron beam irradiator. It is constructed by capacitors of 45 nF and 28.13 nF in the coupling column and capacitors of 18.75 nF in the smoothing column. Working status of the rectifier in high power output condition was analyzed, and the conduction angle of the rectifier was calculated. The power factor (PF) of the SCW circuit has been studied, and the equivalent condensance of the circuit has been derived. Measurements were done for the PF compensation. The surge impact during the short circuit transient process was considered in choosing the protection resistance. Test results showed that design specifications of the power supply were achieved, with the non-load voltage being up to 1.32 MV and the ratio of ripple voltage to output voltage as 9.4%. PMID- 21639548 TI - Ultrathin aluminum sample cans for single crystal inelastic neutron scattering. AB - Single crystal inelastic neutron scattering measurements are often performed using a sample environment for controlling sample temperature. One difficulty associated with this is establishing appropriate thermal coupling from the sample to the temperature controlled portion of the sample environment. This is usually accomplished via a sample can which thermally couples the sample environment to the sample can and the sample can to the sample via an exchange gas. Unfortunately, this can will contribute additional background signal to one's measurement. We present here the design of an ultrathin aluminum sample can based upon established technology for producing aluminum beverage cans. This design minimizes parasitic sample can scattering. Neutron scattering measurements comparing a machined sample can to our beverage can design clearly indicate a large reduction in scattering intensity and texture when using the ultrathin sample can design. We also examine the possibility of using standard commercial beverage cans as sample cans. PMID- 21639549 TI - Note: Development of a high resolution six-degrees-of-freedom optical vibrometer for precision stage. AB - A high resolution six degrees of freedom (6-DOF) optical vibrometer is proposed. 6-DOF vibrations can be simultaneously measured using the proposed optical vibrometer, which reduces measurement time and number. The performance of the proposed vibrometer is verified by experiments. The results show that the accuracy of the proposed optical vibrometer is +/-30 nm/200 nm and +/-0.04 arcsec/0.1 arcsec at 1000 Hz. PMID- 21639550 TI - Note: Non-gain microchannel plate gated framing camera. AB - An x-ray framing camera using a non-gain microchannel plate (MCP) is reported in this article. The advantage of the non-gain MCP is the less transit time spread. The non-gain MCP gated framing camera has four microstrip line cathodes with 6 mm in width. The time domain reflectometry curves of the four microstrip lines are measured, which show that the characteristic impedance of each microstrip line is about 17 Omega. While the photocathode is driven by the gating electrical pulse with width of 125 ps and amplitude of -1.48 kV with -400 V bias, the measured exposure time of this camera is about 72 ps. PMID- 21639551 TI - Note: A novel atomic force microscope fast imaging approach: variable-speed scanning. AB - Imaging speed is one of the key factors limiting atomic force microscope's (AFM) wide applications. To improve its performance, a variable-speed scanning (VSS) method is designed in this note for an AFM. Specifically, in the VSS mode, the scanning speed is tuned online according to the feedback information to properly distribute imaging time along sample surface. Furthermore, some practical mechanism is proposed to determine the best time of moving the AFM tip to the next scanned point. The contrast experiment results show that the VSS method speeds up the imaging rate while ensuring image quality. PMID- 21639552 TI - Note: Radio frequency surface impedance characterization system for superconducting samples at 7.5 GHz. AB - A radio frequency (RF) surface impedance characterization (SIC) system that uses a novel sapphire-loaded niobium cavity operating at 7.5 GHz has been developed as a tool to measure the RF surface impedance of flat superconducting material samples. The SIC system can presently make direct calorimetric RF surface impedance measurements on the central 0.8 cm(2) area of 5 cm diameter disk samples from 2 to 20 K exposed to RF magnetic fields up to 14 mT. To illustrate system utility, we present first measurement results for a bulk niobium sample. PMID- 21639553 TI - Note: Laser ablation technique for electrically contacting a buried implant layer in single crystal diamond. AB - The creation of thin, buried, and electrically conducting layers within an otherwise insulating diamond by annealed ion implantation damage is well known. Establishing facile electrical contact to the shallow buried layer has been an unmet challenge. We demonstrate a new method, based on laser micro-machining (laser ablation), to make reliable electrical contact to a buried implant layer in diamond. Comparison is made to focused ion beam milling. PMID- 21639554 TI - Note: Design of transverse electron gun for electron beam based reactive evaporation system. AB - In this paper design of a 10 kV, 10 kW transverse electron gun, suitable for reactive evaporation, supported by simulation and modeling, is presented. Simulation of the electron beam trajectory helps in locating the emergence aperture after 90 degrees bend and also in designing the crucible on which the beam is finally incident after 270 degrees bend. The dimension of emergence aperture plays a vital role in designing the differential pumping system between the gun chamber and the substrate chamber. Experimental validation is done for beam trajectory by piercing a stainless steel plate at 90 degrees position which is kept above the crucible. PMID- 21639557 TI - Fiber loop ringdown DNA and bacteria sensors. AB - We report a new type of refractive index-based biosensor using a fiber loop ringdown evanescent field (FLRD-EF) sensing scheme, in which the sensing signal is a time constant and detection sensitivity is enhanced by the multipass nature of the ringdown technique. Bulk index-based detections of three different single strand DNAs and one type of bacteria are demonstrated for the FLRD-EF sensors that utilize a partially-etched single mode fiber as the sensor head. Stepwise coating of the sensor head with poly-L-lysine and a probe DNA has enabled surface index-based label-free target DNA sensing. We expect an array of FLRD-EF biosensors to be created, which are superior to counterparts in terms of simplicity, low cost, and high sensitivity. PMID- 21639558 TI - Confocal imaging reveals three-dimensional fine structure difference between ventral and dorsal nerve roots. AB - Peripheral nerve injury repair is one of the most challenging problems in neurosurgery, partially due to lack of knowledge of three-dimensional (3-D) fine structure and organization of peripheral nerves. In this paper, we explored the structures of nerve fibers in ventral and dorsal nerves with a laser scanning confocal microscopy. Thick tissue staining results suggested that nerve fibers have a different 3-D structure in ventral and dorsal nerves, and reconstruction from serial sectioning images showed that in ventral nerves the nerve fibers travel in a winding form, while in dorsal nerves, the nerve fibers form in a parallel cable pattern. These structural differences could help surgeons to differentiate ventral and dorsal nerves in peripheral nerve injury repair, and also facilitate scientists to get a deeper understanding about nerve fiber organization. PMID- 21639559 TI - Multifunctional imaging of human retina and choroid with 1050-nm spectral domain optical coherence tomography at 92-kHz line scan rate. AB - The light source at ~1-MUm wavelength is attractive for enhanced imaging depth in retinal optical coherence tomography (OCT). In this paper, we report on a 1050-nm spectral domain OCT system, combined with optical microangiography that operates at a 92-kHz line scan rate for multifunctional imaging of the human eye, delivering the volumetric imaging of microstructure and microvasculature within retina and choroid. PMID- 21639560 TI - Rapid confocal imaging of large areas of excised tissue with strip mosaicing. AB - Imaging large areas of tissue rapidly and with high resolution may enable rapid pathology at the bedside. The limited field of view of high-resolution microscopes requires the merging of multiple images that are taken sequentially to cover a large area. This merging or mosaicing of images requires long acquisition and processing times, and produces artifacts. To reduce both time and artifacts, we developed a mosaicing method on a confocal microscope that images morphology in large areas of excised tissue with sub-cellular detail. By acquiring image strips with aspect ratios of 10:1 and higher (instead of the standard ~1:1) and "stitching" them in software, our method images 10 * 10 mm(2) area of tissue in about 3 min. This method, which we call "strip mosaicing," is currently three times as fast as our previous method. PMID- 21639561 TI - Special section guest editorial: Photonics and nanotechnology in biophysics and biomedical research. PMID- 21639562 TI - Survey on indirect optical manipulation of cells, nucleic acids, and motor proteins. AB - Optical tweezers have emerged as a promising technique for manipulating biological objects. Instead of direct laser exposure, more often than not, optically-trapped beads are attached to the ends or boundaries of the objects for translation, rotation, and stretching. This is referred to as indirect optical manipulation. In this paper, we utilize the concept of robotic gripping to explain the different experimental setups which are commonly used for indirect manipulation of cells, nucleic acids, and motor proteins. We also give an overview of the kind of biological insights provided by this technique. We conclude by highlighting the trends across the experimental studies, and discuss challenges and promising directions in this domain of active current research. PMID- 21639563 TI - Effect of polyethylene glycol coatings on uptake of indocyanine green loaded nanocapsules by human spleen macrophages in vitro. AB - Near-infrared (NIR) optically active nanoparticles are promising exogenous chromophores for applications in medical imaging and phototherapy. Since nanoparticles can be rapidly eliminated from the body by cells of the reticuloendothelial system, a thriving strategy to increase their blood circulation time is through surface modification with polyethylene glycol (PEG). We constructed polymeric nanocapsules loaded with indocyanine green (ICG), an FDA approved NIR dye, and coated with aldehyde-terminated PEG. Using optical absorbance spectroscopy and flow cytometry, we investigated the effect of PEG coating and molecular weight (MW) of PEG [5000 and 30,000 Daltons (Da)] on the phagocytic content of human spleen macrophages incubated with ICG-containing nanocapsules (ICG-NCs) between 15 to 360 min. Our results indicate that surface coating with PEG is an effective method to reduce the phagocytic content of ICG NCs within macrophages for at least up to 360 min of incubation time. Coating the surface of ICG-NCs with the low MW PEG results in lower phagocytic content of ICG NCs within macrophages for at least up to 60 min of incubation time as compared to ICG-NCs coated with the high MW PEG. Surface coating of ICG-NCs with PEG is a promising approach to prolong vasculature circulation time of ICG for NIR imaging and phototherapeutic applications. PMID- 21639564 TI - Optical properties of breast tumor phantoms containing carbon nanotubes and nanohorns. AB - The degree by which optical properties of tumors are altered following introduction of carbon nanotubes (CNTs) of varying concentration and type is poorly understood, making it difficult to predict the impact of CNT inclusion on the photothermal response to laser therapies. Optical properties were measured of phantoms representative of breast tumor tissue incorporated with multiwalled carbon nanotubes (MWNTs), single-walled carbon nanotubes (SWNTs), and single walled carbon nanohorns (SWNHs) of varying concentration (0.01-0.1 mg/ml). Tissue phantoms were made from sodium alginate (3 g/ml) incorporated with polystyrene microbeads (3 MUm diam and 1 mg/ml) and talc-France powder (40 mg/ml). Absorption (MU(a)) and reduced scattering (MU's) coefficients of phantoms containing CNTs were determined by the inverse adding-doubling algorithm for the wavelength range of 400-1300 nm. Optical properties of phantoms without CNTs were in the range of MU(a) = 1.04-0.06 mm(-1) and MU's' = 0.05-0.07 mm(-1) at a wavelength of 900 nm, which corresponds with published data for human breast tumor tissue. Incorporating MWNTs, SWNTs, and SWNHs in phantoms with a concentration of 0.1 mg/ml increased (MU(a)) by 20- to 30-fold, 5- to 6-fold, and 9- to 14-fold, respectively, for the wavelength range of 800-1100 nm with minimal change in MU's (1.2- to 1.3-fold). Introduction of CNTs into tissue phantoms increased absorption, providing a means to enhance photothermal therapy. PMID- 21639565 TI - Multiphoton microscopy study of the morphological and quantity changes of collagen and elastic fiber components in keloid disease. AB - Multiphoton microscopy was used to study the extracellular matrix of keloid at the molecular level without tissue fixation and staining. Direct imaging of collagen and elastin was achieved by second harmonic generation and two-photon excited fluorescence, respectively. The morphology and quantity of collagen and elastin in keloid were characterized and quantitatively analyzed in comparison to normal skin. The study demonstrated that in keloid, collagen content increased in both the upper dermis and the deep dermis, while elastin mostly showed up in the deep dermis and its quantity is higher compared to normal skin. This suggests the possibility that abnormal fibroblasts synthesized an excessive amount of collagen and elastin at the beginning of keloid formation, corresponding to the observed deep dermis, while after a certain time point, the abnormal fibroblast produced mostly collagen, corresponding to the observed upper dermis. The morphology of collagen and elastin in keloid was disrupted and presented different variations. In the deep dermis, elastic fibers showed node structure, while collagen showed obviously regular gaps between adjacent bundles. In the upper dermis, collagen bundles aligned in a preferred direction, while elastin showed as sparse irregular granules. This new molecular information provided fresh insight about the development process of keloid. PMID- 21639566 TI - Combined optical tweezers and laser dissector for controlled ablation of functional connections in neural networks. AB - Regeneration of functional connectivity within a neural network after different degrees of lesion is of utmost clinical importance. To test pharmacological approaches aimed at recovering from a total or partial damage of neuronal connections within a circuit, it is necessary to develop a precise method for controlled ablation of neuronal processes. We combined a UV laser microdissector to ablate neural processes in vitro at single neuron and neural network level with infrared holographic optical tweezers to carry out force spectroscopy measurements. Simultaneous force spectroscopy, down to the sub-pico-Newton range, was performed during laser dissection to quantify the tension release in a partially ablated neurite. Therefore, we could control and measure the damage inflicted to an individual neuronal process. To characterize the effect of the inflicted injury on network level, changes in activity of neural subpopulations were monitored with subcellular resolution and overall network activity with high temporal resolution by concurrent calcium imaging and microelectrode array recording. Neuronal connections have been sequentially ablated and the correlated changes in network activity traced and mapped. With this unique combination of electrophysiological and optical tools, neural activity can be studied and quantified in response to controlled injury at the subcellular, cellular, and network level. PMID- 21639567 TI - Dual-mode imaging with radiolabeled gold nanorods. AB - Many nanoparticle contrast agents have difficulties with deep tissue and near bone imaging due to limited penetration of visible photons in the body and mineralized tissues. We are looking into the possibility of mediating this problem while retaining the capabilities of the high spatial resolution associated with optical imaging. As such, the potential combination of emerging photoacoustic imaging and nuclear imaging in monitoring of antirheumatic drug delivery by using a newly developed dual-modality contrast agent is investigated. The contrast agent is composed of gold nanorods (GNRs) conjugated to the tumor necrosis factor (TNF-alpha) antibody and is subsequently radiolabeled by (125)I. ELISA experiments designed to test TNF-alpha binding are performed to prove the specificity and biological activity of the radiolabeled conjugated contrast agent. Photoacoustic and nuclear imaging are performed to visualize the distribution of GNRs in articular tissues of the rat tail joints in situ. Findings from the two imaging modalities correspond well with each other in all experiments. Our system can image GNRs down to a concentration of 10 pM in biological tissues and with a radioactive label of 5 MUCi. This study demonstrates the potential of combining photoacoustic and nuclear imaging modalities through one targeted contrast agent for noninvasive monitoring of drug delivery as well as deep and mineralized tissue imaging. PMID- 21639568 TI - Mechanical design of a power-adjustable spectacle lens frame. AB - Power-adjustable spectacle lenses, based on the Alvarez-Lohmann principle, can be used to provide affordable spectacles for subjective refractive errors measurement and its correction. A new mechanical frame has been designed to maximize the advantages of this technology. The design includes a mechanism to match the interpupillary distance with that of the optical centers of the lenses. The frame can be manufactured using low cost plastic injection molding techniques. A prototype has been built to test the functioning of this mechanical design. PMID- 21639569 TI - Low-noise small-size microring ultrasonic detectors for high-resolution photoacoustic imaging. AB - Small size polymer microring resonators have been exploited for photoacoustic (PA) imaging. To demonstrate the advantages of the wide acceptance angle of ultrasound detection of small size microrings, photoacoustic tomography (PAT), and delay-and-sum beamforming PA imaging was conducted. In PAT, we compared the imaging quality using different sizes of detectors with similar noise-equivalent pressures and the same wideband response: 500 MUm hydrophone and 100, 60, and 40 MUm microrings. The results show significantly improved imaging contrast and high resolution over the whole imaging region using smaller size detectors. The uniform high resolution in PAT imaging using 40 MUm microrings indicates the potential to resolve microvasculature over a large imaging region. The improved lateral resolution of two-dimensional and three-dimensional delay-and-sum beamforming PA imaging using a synthetic array demonstrate another advantageous application of small microrings. The small microrings can also be applied to other ultrasound-related imaging applications. PMID- 21639570 TI - Depth-correction algorithm that improves optical quantification of large breast lesions imaged by diffuse optical tomography. AB - Optical quantification of large lesions imaged with diffuse optical tomography in reflection geometry is depth dependence due to the exponential decay of photon density waves. We introduce a depth-correction method that incorporates the target depth information provided by coregistered ultrasound. It is based on balancing the weight matrix, using the maximum singular values of the target layers in depth without changing the forward model. The performance of the method is evaluated using phantom targets and 10 clinical cases of larger malignant and benign lesions. The results for the homogenous targets demonstrate that the location error of the reconstructed maximum absorption coefficient is reduced to the range of the reconstruction mesh size for phantom targets. Furthermore, the uniformity of absorption distribution inside the lesions improve about two times and the median of the absorption increases from 60 to 85% of its maximum compared to no depth correction. In addition, nonhomogenous phantoms are characterized more accurately. Clinical examples show a similar trend as the phantom results and demonstrate the utility of the correction method for improving lesion quantification. PMID- 21639571 TI - Semiautomated analysis of optical coherence tomography crystalline lens images under simulated accommodation. AB - Presbyopia is an age related, gradual loss of accommodation, mainly due to changes in the crystalline lens. As part of research efforts to understand and cure this condition, ex vivo, cross-sectional optical coherence tomography images of crystalline lenses were obtained by using the Ex-Vivo Accommodation Simulator (EVAS II) instrument and analyzed to extract their physical and optical properties. Various filters and edge detection methods were applied to isolate the edge contour. An ellipse is fitted to the lens outline to obtain central reference point for transforming the pixel data into the analysis coordinate system. This allows for the fitting of a high order equation to obtain a mathematical description of the edge contour, which obeys constraints of continuity as well as zero to infinite surface slopes from apex to equator. Geometrical parameters of the lens were determined for the lens images captured at different accommodative states. Various curve fitting functions were developed to mathematically describe the anterior and posterior surfaces of the lens. Their differences were evaluated and their suitability for extracting optical performance of the lens was assessed. The robustness of these algorithms was tested by analyzing the same images repeated times. PMID- 21639572 TI - Comparison of retinal thickness by Fourier-domain optical coherence tomography and OCT retinal image analysis software segmentation analysis derived from Stratus optical coherence tomography images. AB - PURPOSE: To compare thickness measurements between Fourier-domain optical coherence tomography (FD-OCT) and time-domain OCT images analyzed with a custom built OCT retinal image analysis software (OCTRIMA). METHODS: Macular mapping (MM) by StratusOCT and MM5 and MM6 scanning protocols by an RTVue-100 FD-OCT device are performed on 11 subjects with no retinal pathology. Retinal thickness (RT) and the thickness of the ganglion cell complex (GCC) obtained with the MM6 protocol are compared for each early treatment diabetic retinopathy study (ETDRS) like region with corresponding results obtained with OCTRIMA. RT results are compared by analysis of variance with Dunnett post hoc test, while GCC results are compared by paired t-test. RESULTS: A high correlation is obtained for the RT between OCTRIMA and MM5 and MM6 protocols. In all regions, the StratusOCT provide the lowest RT values (mean difference 43 +/- 8 MUm compared to OCTRIMA, and 42 +/ 14 MUm compared to RTVue MM6). All RTVue GCC measurements were significantly thicker (mean difference between 6 and 12 MUm) than the GCC measurements of OCTRIMA. CONCLUSION: High correspondence of RT measurements is obtained not only for RT but also for the segmentation of intraretinal layers between FD-OCT and StratusOCT-derived OCTRIMA analysis. However, a correction factor is required to compensate for OCT-specific differences to make measurements more comparable to any available OCT device. PMID- 21639573 TI - Real-time snapshot hyperspectral imaging endoscope. AB - Hyperspectral imaging has tremendous potential to detect important molecular biomarkers of early cancer based on their unique spectral signatures. Several drawbacks have limited its use for in vivo screening applications: most notably the poor temporal and spatial resolution, high expense, and low optical throughput of existing hyperspectral imagers. We present the development of a new real-time hyperspectral endoscope (called the image mapping spectroscopy endoscope) based on an image mapping technique capable of addressing these challenges. The parallel high throughput nature of this technique enables the device to operate at frame rates of 5.2 frames per second while collecting a (x, y, lambda) datacube of 350 * 350 * 48. We have successfully imaged tissue in vivo, resolving a vasculature pattern of the lower lip while simultaneously detecting oxy-hemoglobin. PMID- 21639574 TI - Multiexposure laser speckle contrast imaging of the angiogenic microenvironment. AB - We report the novel use of laser speckle contrast imaging (LSCI) at multiple exposure times (meLSCI) for enhanced in vivo imaging of the microvascular changes that accompany angiogenesis. LSCI is an optical imaging technique that can monitor blood vessels and the flow therein at a high spatial resolution without requiring the administration of an exogenous contrast agent. LSCI images are obtained under red (632 nm) laser illumination at seven exposure times (1-7 ms) and combined using a curve-fitting approach to obtain high-resolution meLSCI images of the rat brain vasculature. To evaluate enhancement in in vivo imaging performance, meLSCI images are statistically compared to individual LSCI images obtained at a single exposure time. We find that meLSCI reduced the observed variability in the LSCI-based blood-flow estimates by 30% and improved the contrast-to-noise ratio in regions with high microvessel density by 41%. The ability to better distinguish microvessels, makes meLSCI uniquely suited to longitudinal imaging of changes in the vascular microenvironment induced by pathological angiogenesis. We demonstrate this utility of meLSCI by sequentially monitoring, over days, the microvascular changes that accompany wound healing in a mouse ear model. PMID- 21639575 TI - Extended scan depth optical coherence tomography for evaluating ocular surface shape. AB - Spectral domain optical coherence tomography (SD-OCT) with extended scan depth makes it possible for quantitative measurement of the entire ocular surface shape. We proposed a novel method for ocular surface shape measurement using a custom-built anterior segment SD-OCT, which will serve on the contact lens fitting. A crosshair alignment system was applied to reduce the misalignment and tilting of the eye. An algorithm was developed to automatically segment the ocular surface. We also described the correction of the image distortion from the segmented dataset induced by the nontelecentric scanning system and tested the accuracy and repeatability. The results showed high accuracy of SD-OCT in measuring a bicurved test surface with a maximum height error of 17.4 MUm. The repeatability of in vivo measurement was also good. The standard deviations of the height measurement within a 14-mm wide range were all less than 35 MUm. This work demonstrates the feasibility of using extended depth SD-OCT to perform noninvasive evaluation of the ocular surface shape. PMID- 21639576 TI - Silver nanostructure sensing platform for maximum-contrast fluorescence cell imaging. AB - We present herein a silver nanostructure-assisted sensing platform which consists of a combined structure of Ag nanowire (NW) and nanodot (ND) array. Highly enhanced fluorescence from fluorophore is attributed to a strongly coupled optical near-field interaction between proximately located Ag NW and NDs. We obtained enhanced fluorescence intensity with up to 140 folds, as contrasted from background intensity, reaching a theoretical maximum value. On the other hand, fluorescence lifetime was greatly reduced to 0.27 ns (from 2.17 ns for the same fluorophores without nanostructure). This novel platform can be a promising utility for optical imaging and labeling of biological systems with a great sensitivity. PMID- 21639577 TI - Measuring changes in chemistry, composition, and molecular structure within hair fibers by infrared and Raman spectroscopic imaging. AB - Spatially resolved infrared (IR) and Raman images are acquired from human hair cross sections or intact hair fibers. The full informational content of these spectra are spatially correlated to hair chemistry, anatomy, and structural organization through univariate and multivariate data analysis. Specific IR and Raman images from untreated human hair describing the spatial dependence of lipid and protein distribution, protein secondary structure, lipid chain conformational order, and distribution of disulfide cross-links in hair protein are presented in this study. Factor analysis of the image plane acquired with IR microscopy in hair sections, permits delineation of specific micro-regions within the hair. These data indicate that both IR and Raman imaging of molecular structural changes in a specific region of hair will prove to be valuable tools in the understanding of hair structure, physiology, and the effect of various stresses upon its integrity. PMID- 21639578 TI - Enhanced two-channel nonlinear imaging by a highly polarized supercontinuum light source generated from a nonlinear photonic crystal fiber with two zero-dispersion wavelengths. AB - Real-time monitoring the variation of chlorophyll distributions and cellular structures in leaves during plant growth provides important information for understanding the physiological statuses of plants. Two-photon-excited autofluorescence imaging and second harmonic generation imaging of leaves can be used for monitoring the nature intrinsic fluorophores distribution and cellular structures of leaves by the use of the near-infrared region of light which has minimal light absorption by endogenous molecules and thus increases tissue penetration. However, the two-photon absorption peak of intrinsic fluorophores of a ficus benjamina leaf is 50 nm away from the second harmonic generation excitation wavelength, which cannot be effectively excited by a femtosecond laser beam with one central wavelength. This paper shows that a highly polarized supercontinuum light generated from a birefringent nonlinear photonic crystal fiber with two zero-dispersion wavelengths can effectively excite two-photon autofluorescence as well as second harmonic generation signals for simultaneously monitoring intrinsic fluorophore distributions and non-centrosymmetric structures of leaves. PMID- 21639579 TI - Coagulation dynamics of a blood sample by multiple scattering analysis. AB - We report a new technique to measure coagulation dynamics on whole-blood samples. The method relies on the analysis of the speckle figure resulting from a whole blood sample mixed with coagulation reagent and introduced in a thin chamber illuminated with a coherent light. A dynamic study of the speckle reveals a typical behavior due to coagulation. We compare our measured coagulation times to a reference method obtained in a medical laboratory. PMID- 21639580 TI - Analysis of human nails by laser-induced breakdown spectroscopy. AB - Laser-induced breakdown spectroscopy (LIBS) is applied to analyze human fingernails using nanosecond laser pulses. Measurements on 45 nail samples are carried out and 14 key species are identified. The elements detected with the present system are: Al, C, Ca, Fe, H, K, Mg, N, Na, O, Si, Sr, Ti as well as CN molecule. Sixty three emission lines have been identified in the spectrum that are dominated by calcium lines. A discriminant function analysis is used to discriminate among different genders and age groups. This analysis demonstrates efficient discrimination among these groups. The mean concentration of each element is compared between different groups. Correlation between concentrations of elements in fingernails is calculated. A strong correlation is found between sodium and potassium while calcium and magnesium levels are inversely correlated. A case report on high levels of sodium and potassium in patients with hyperthyroidism is presented. It is shown that LIBS could be a promising technique for the analysis of nails and therefore identification of health problems. PMID- 21639581 TI - Terahertz sensing in corneal tissues. AB - This work introduces the potential application of terahertz (THz) sensing to the field of ophthalmology, where it is uniquely suited due to its nonionizing photon energy and high sensitivity to water content. Reflective THz imaging and spectrometry data are reported on ex-vivo porcine corneas prepared with uniform water concentrations using polyethylene glycol (PEG) solutions. At 79% water concentration by mass, the measured reflectivity of the cornea was 20.4%, 14.7%, 11.7%, 9.6%, and 7.4% at 0.2, 0.4, 0.6, 0.8, and 1 THz, respectively. Comparison of nine corneas hydrated from 79.1% to 91.5% concentration by mass demonstrated an approximately linear relationship between THz reflectivity and water concentration, with a monotonically decreasing slope as the frequency increases. The THz-corneal tissue interaction is simulated with a Bruggeman model with excellent agreement. THz applications to corneal dystrophy, graft rejection, and refractive surgery are examined from the context of these measurements. PMID- 21639582 TI - Regional tissue oxygen saturation: comparability and reproducibility of different devices. AB - Comparability and reproducibility of different near-infrared spectroscopy devices measuring regional tissue oxygen saturation remain poor. Aim of the present study was to compare values and reproducibility of cerebral/peripheral "tissue oxygenation-index" (TOI; NIRO 300, Hamamatsu((r)), Japan) with cerebral/peripheral "regional-oxygen-saturation" (rSO; INVOS5100, Somanetics((r)), USA), and to analyze the influence of quality criteria. METHODS: cTOI and crSO2 were measured on the left forehead, pTOI and prSO2 were measured on the left calf. To analyse reproducibility, optodes were reapplied five times. A quality criterion was introduced for cTOI, crSO2 and prSO2. For pTOI quality criteria were introduced in combination with a venous occlusion technique. RESULTS: Cerebral measurements were performed in 37 neonates. cTOI (72.7+/-6.2%) was lower than crSO2 (83.3+/-5.8%) (p < 0.001). The mean difference between cTOI and crSO2 was 10%. Mean standard deviations of cTOI and crSO2 were similar (cTOI: 4.9+/-3.6; crSO2: 4.5+/-2.6). Peripheral measurements were performed in 39 neonates. pTOI (66.0+/-7.9%) was lower than prSO2 (82.0+/-7.0%)(p < 0.001). The mean difference between pTOI and prSO2 was 15%. Mean standard deviations of pTOI (3.7+/-2.6%) were lower than of prSO2 (5.0+/-3.0%) (p = 0.047). CONCLUSION: TOI values were significantly lower than rSO2 values, in cerebral and peripheral measurements. Reproducibility was higher for pTOI than for prSO2. PMID- 21639583 TI - Spectroscopic signature of mouse embryonic stem cell-derived hepatocytes using synchrotron Fourier transform infrared microspectroscopy. AB - Stem cell-based therapy for liver regeneration has been proposed to overcome the persistent shortage in the supply of suitable donor organs. A requirement for this to succeed is to find a rapid method to detect functional hepatocytes, differentiated from embryonic stem cells. We propose Fourier transform infrared (FTIR) microspectroscopy as a versatile method to identify the early and last stages of the differentiation process leading to the formation of hepatocytes. Using synchrotron-FTIR microspectroscopy, the means of identifying hepatocytes at the single-cell level is possible and explored. Principal component analysis and subsequent partial least-squares (PLS) discriminant analysis is applied to distinguish endoderm induction from hepatic progenitor cells and matured hepatocyte-like cells. The data are well modeled by PLS with endoderm induction, hepatic progenitor cells, and mature hepatocyte-like cells able to be discriminated with very high sensitivity and specificity. This method provides a practical tool to monitor endoderm induction and has the potential to be applied for quality control of cell differentiation leading to hepatocyte formation. PMID- 21639584 TI - Automatic cytometric device using multiple wavelength excitations. AB - Precise identification of eosinophils, basophils, and specific subpopulations of blood cells (B lymphocytes) in an unconventional automatic hematology analyzer is demonstrated. Our specific apparatus mixes two excitation radiations by means of an acousto-optics tunable filter to properly control fluorescence emission of phycoerythrin cyanin 5 (PC5) conjugated to antibodies (anti-CD20 or anti-CRTH2) and Thiazole Orange. This way our analyzer combining techniques of hematology analysis and flow cytometry based on multiple fluorescence detection, drastically improves the signal to noise ratio and decreases the spectral overlaps impact coming from multiple fluorescence emissions. PMID- 21639585 TI - Image-guided genomic analysis of tissue response to laser-induced thermal stress. AB - The cytoprotective response to thermal injury is characterized by transcriptional activation of "heat shock proteins" (hsp) and proinflammatory proteins. Expression of these proteins may predict cellular survival. Microarray analyses were performed to identify spatially distinct gene expression patterns responding to thermal injury. Laser injury zones were identified by expression of a transgene reporter comprised of the 70 kD hsp gene and the firefly luciferase coding sequence. Zones included the laser spot, the surrounding region where hsp70-luc expression was increased, and a region adjacent to the surrounding region. A total of 145 genes were up-regulated in the laser irradiated region, while 69 were up-regulated in the adjacent region. At 7 hours the chemokine Cxcl3 was the highest expressed gene in the laser spot (24 fold) and adjacent region (32 fold). Chemokines were the most common up-regulated genes identified. Microarray gene expression was successfully validated using qRT- polymerase chain reaction for selected genes of interest. The early response genes are likely involved in cytoprotection and initiation of the healing response. Their regulatory elements will benefit creating the next generation reporter mice and controlling expression of therapeutic proteins. The identified genes serve as drug development targets that may prevent acute tissue damage and accelerate healing. PMID- 21639586 TI - Feasibility of intrafraction whole-body motion tracking for total marrow irradiation. AB - With image-guided tomotherapy, highly targeted total marrow irradiation (TMI) has become a feasible alternative to conventional total body irradiation. The uncertainties in patient localization and intrafraction motion of the whole body during hour-long TMI treatment may pose a risk to the safety and accuracy of targeted radiation treatment. The feasibility of near-infrared markers and optical tracking system (OTS) is accessed along with a megavoltage scanning system of tomotherapy. Three near-infrared markers placed on the face of a rando phantom are used to evaluate the capability of OTS in measuring changes in the markers' positions as the rando is moved in the translational direction. The OTS is also employed to determine breathing motion related changes in the position of 16 markers placed on the chest surface of human volunteers. The maximum uncertainty in locating marker position with the OTS is 1.5 mm. In the case of normal and deep breathing motion, the maximum marker position change is observed in anterior-posterior direction with the respective values of 4 and 12 mm. The OTS is able to measure surface changes due to breathing motion. The OTS may be optimized to monitor whole body motion during TMI to increase the accuracy of treatment delivery and reduce the radiation dose to the lungs. PMID- 21639587 TI - Determination of the tumor tissue optical properties during and after photodynamic therapy using inverse Monte Carlo method and double integrating sphere between 350 and 1000 nm. AB - Photodynamic therapy (PDT) efficacy depends on the amount of light distribution within the tissue. However, conventional PDT does not consider the laser irradiation dose during PDT. The optical properties of biological tissues (absorption coefficient MU(a), reduced scattering coefficient MU's), anisotropy factor g, refractive index, etc.) help us to recognize light propagation through the tissue. The goal of this paper is to acquire the knowledge of the light propagation within tissue during and after PDT with the optical property of PDT performed mouse tumor tissue. The optical properties of mouse tumor tissues were evaluated using a double integrating sphere setup and the algorithm based on the inverse Monte Carlo method in the wavelength range from 350 to 1000 nm. During PDT, the MU(a) and MU's were not changed after 1 and 5 min of irradiation. After PDT, the MU's in the wavelength range from 600 to 1000 nm increased with the passage of time. For seven days after PDT, the MU's increased by 1.7 to 2.0 times, which results in the optical penetration depth decreased by 1.4 to 1.8 times. To ensure an effective procedure, the adjustment of laser parameters for the decreasing penetration depth is recommended for the re-irradiation of PDT. PMID- 21639589 TI - Critical perspective on the consequences of the limited availability of kinetic data in metabolic dynamic modelling. AB - Detailed kinetic models at the network reaction level are usually constructed using enzymatic mechanistic rate equations and the associated kinetic parameters. However, during the cellular life cycle thousands of different reactions occur, which makes it very difficult to build a detailed large-scale ldnetic model. In this work, we provide a critical overview of specific limitations found during the reconstruction of the central carbon metabolism dynamic model from E. coli (based on kinetic data available). In addition, we provide clues that will hopefully allow the systems biology community to more accurately construct metabolic dynamic models in the future. The difficulties faced during the construction of dynamic models are due not only to the lack of kinetic information but also to the fact that some data are still not curated. We hope that in the future, with the standardization of the in vitro enzyme protocols the approximation of in vitro conditions to the in vivo ones, it will be possible to integrate the available kinetic data into a complete large scale model. We also expect that collaborative projects between modellers and biologists will provide valuable kinetic data and permit the exchange of important information to solve most of these issues. PMID- 21639590 TI - Cell population modelling describes intrinsic heterogeneity: a case study for hematopoietic stem cells. AB - The control of stem cell properties during in vitro expansion is of paramount importance for their clinical use. According to Food and Drug Administration (FDA) guidelines, phenotypic heterogeneity is a critical aspect influencing therapeutic response. Even if the authors ability to reduce heterogeneity were limited, the sources from which it arises should be well understood for safe clinical applications. The aim of this work was to describe theoretically the intrinsic cell population heterogeneity that is present even when cells are cultured in a perfectly homogeneous environment. A bivariate population balance model is developed to account for the heterogeneity in the number of receptors and receptor-ligand complexes per cell, and is coupled with a ligand conservation equation. As a case study, the model is applied to the hematopoietic stem cell expansion, considering the c-Kit receptor and stem cell factor pair. Results show the dependence of intrinsic heterogeneity from ligand concentration and the kinetics of its administration. By tracking the cell generations within the total population, the authors highlight intra- and an inter-generational contributions to total population heterogeneity. In terms of dimensionless variables, intrinsic heterogeneity is dependent on the ratio of the characteristic time of cell division to that needed by a newborn cell to reach its single-cell steady state. [Includes supplementary material]. PMID- 21639591 TI - Exploring phospholipase C-coupled Ca(2+) signalling networks using Boolean modelling. AB - In this study, the authors explored the utility of a descriptive and predictive bionetwork model for phospholipase C-coupled calcium signalling pathways, built with non-kinetic experimental information. Boolean models generated from these data yield oscillatory activity patterns for both the endoplasmic reticulum resident inositol-1,4,5-trisphosphate receptor (IP(3)R) and the plasma-membrane resident canonical transient receptor potential channel 3 (TRPC3). These results are specific as randomisation of the Boolean operators ablates oscillatory pattern formation. Furthermore, knock-out simulations of the IP(3)R, TRPC3 and multiple other proteins recapitulate experimentally derived results. The potential of this approach can be observed by its ability to predict previously undescribed cellular phenotypes using in vitro experimental data. Indeed, our cellular analysis of the developmental and calcium-regulatory protein, DANGER1a, confirms the counter-intuitive predictions from our Boolean models in two highly relevant cellular models. Based on these results, the authors theorise that with sufficient legacy knowledge and/or computational biology predictions, Boolean networks can provide a robust method for predictive modelling of any biological system. [Includes supplementary material]. PMID- 21639592 TI - Networks for systems biology: conceptual connection of data and function. AB - The purpose of this study is to survey the use of networks and network-based methods in systems biology. This study starts with an introduction to graph theory and basic measures allowing to quantify structural properties of networks. Then, the authors present important network classes and gene networks as well as methods for their analysis. In the last part of this study, the authors review approaches that aim at analysing the functional organisation of gene networks and the use of networks in medicine. In addition to this, the authors advocate networks as a systematic approach to general problems in systems biology, because networks are capable of assuming multiple roles that are very beneficial connecting experimental data with a functional interpretation in biological terms. PMID- 21639593 TI - Effects of saturation and enzyme limitation in feedforward adaptive signal transduction. AB - In this study, the author examines the effects of saturation and enzyme limitation in temporal and spatial signal transduction in a generic feedforward adaptive module. The feedforward module encompasses a range of temporal and spatial signal processing, and this study systematically examines the effect of enzyme limitation/saturating effects in each of the feedforward pathways, and their interplay. It is found that this saturation makes the adaptation inexact, and this effect is more pronounced for higher levels of input signals. Further, it has a very significant role in affecting the temporal dynamics of this module. In examining the role of saturation in the module response to static gradients, the author finds that in certain cases, saturation can completely alter the gradient response. The author examines various aspects of the response systematically using analytical methods and simulations. Overall the author studies a framework and basis for examining and understanding the roles of saturating effects in multiple pathways involved in adaptive responses and sheds light on the relationship and connection between exact and inexact adaptation. PMID- 21639594 TI - Mathematical modelling of negative feedback regulation by carboxyltransferase. AB - Acetyl-CoA carboxylase catalyses the first committed step in fatty acid synthesis in all organisms. The chemistry is accomplished in two half-reactions: activation of biotin via carboxylation by biotin carboxylase, followed by the carboxyltransferase-catalysed transfer of the carboxyl moiety from carboxybiotin to acetyl-CoA to generate malonyl-CoA. The Escherichia coli form of the carboxyltransferase subunit was recently found to regulate its own activity and expression by binding its own mRNA. By binding acetyl-CoA or the mRNA encoding its own subunits, carboxyltransferase is able to sense the metabolic state of the cell and attenuate its own translation and enzymatic activity using a negative feedback mechanism. Here, the network of these interactions is modelled mathematically with a set of non-linear differential equations. Numerical simulations of the model show that it qualitatively and quantitatively agrees with the experimental results for both inhibition of carboxyltransferase by mRNA and attenuation of translation. The modelling of the autoregulatory function of carboxyltransferase confirms that it is more than isolated interactions, but functions as a single dynamic system. PMID- 21639595 TI - Gender differences in combat-related stressors and their association with postdeployment mental health in a nationally representative sample of U.S. OEF/OIF veterans. AB - Though the broader literature suggests that women may be more vulnerable to the effects of trauma exposure, most available studies on combat trauma have relied on samples in which women's combat exposure is limited and analyses that do not directly address gender differences in associations between combat exposure and postdeployment mental health. Female service members' increased exposure to combat in Afghanistan and Iraq provides a unique opportunity to evaluate gender differences in different dimensions of combat-related stress and associated consequence for postdeployment mental health. The current study addressed these research questions in a representative sample of female and male U.S. veterans who had returned from deployment to Afghanistan or Iraq within the previous year. As expected, women reported slightly less exposure than men to most combat related stressors, but higher exposure to other stressors (i.e., prior life stress, deployment sexual harassment). No gender differences were observed in reports of perceived threat in the war zone. Though it was hypothesized that combat-related stressors would demonstrate stronger negative associations with postdeployment mental health for women, only one of 16 stressor * gender interactions achieved statistical significance and an evaluation of the clinical significance of these interactions revealed that effects were trivial. Results suggest that female Operation Enduring Freedom/Operation Iraqi Freedom service members may be as resilient to combat-related stress as men. Future research is needed to evaluate gender differences in the longer-term effects of combat exposure. PMID- 21639596 TI - The effects of alcohol and dosage-set on risk-seeking behavior in groups and individuals. AB - A great deal of risky activity occurs in social contexts, yet only recently have studies begun to examine the impact of drinking on risk-seeking behavior in groups. The present study sought to extend this work by examining both pharmacological and expectancy (dosage-set) effects of drinking. In addition, by using a much larger sample than in prior studies we aimed to increase the power to examine how drinking affects the decision-making process (i.e., Does the initial proposed decision stand, or does it shift during discussion to a safer or riskier final decision?). Seven hundred twenty unacquainted social drinkers (half female) were randomly assigned to three-person groups that consumed alcohol (0.82 g/kg males; 0.74 g/kg females), a placebo, or a no-alcohol control beverage. After drinking, participants decided whether to complete a 30-min questionnaire battery (the less risky choice) or toss a coin and, pending the outcome of that toss, complete either no questionnaires or a 60-min battery (the riskier choice). Neither drinking nor believing one had been drinking affected the decision to toss the coin when participants deliberated in isolation. In contrast, when the decision occurred in a group context, groups led to believe they were drinking alcohol (i.e., groups administered alcohol or placebo beverages) were significantly more likely than groups knowing they had consumed a nonalcoholic beverage (i.e., groups administered a no-alcohol control beverage) to choose the coin toss. Results extend prior findings highlighting the effects of alcohol dosage-set in social contexts. PMID- 21639597 TI - Event-specific drinking among college students. AB - College represents a period of risk for heavy drinking and experiencing unwanted consequences associated with drinking. Previous research has identified specific events, including holidays (e.g., New Years), school breaks (e.g., Spring Break) and personally relevant events (e.g., 21st birthdays), that are associated with elevated risk of heavy drinking and negative alcohol-related consequences. The systematic evaluation of relative risk offers insights into event-specific drinking and an empirical basis upon which to consider allocation of limited prevention resources. Thus, the purpose of the present study was to provide a comparative index of drinking across a wide range of holidays and compare holiday drinking to 21st birthday drinking. Participants were 1,124 students (55% female) who had turned 21 within the previous three weeks in 2008 and provided 90-day retrospective reports of their drinking using the Timeline Follow-back. Results based on a hurdle mixed model for blood alcohol content revealed several holidays that stand out for elevated drinking, including New Year's Eve and July 4th, whereas other holidays appear more similar to weekend drinking, such as Spring Break (approximately last week of March) and graduation (mid-June). Drinking on holidays or special days was substantially lower than drinking on 21st birthdays. Results are discussed in terms of practical applications for targeted intervention efforts on college campuses toward specific events where elevated drinking is known to occur. PMID- 21639598 TI - Rose-colored beer goggles: the relation between experiencing alcohol consequences and perceived likelihood and valence. AB - Although experiencing alcohol-related consequences has some influence on future drinking, this effect may be stronger based on the degree to which the consequence is viewed as positive versus negative, either by the individual or predefined by researchers. This study explored the relationship between experiencing positive and negative alcohol-related consequences and college students' perceptions of how likely those consequences were to occur in the future (i.e., likelihood), and their view of how positive or negative experiencing those consequences would be if they did experience them as a result of drinking (i.e., valence). Data were collected from 491 college students (mean age = 19.26; 56.4% female; 55.0% Caucasian; 33.2% Asian/Pacific Islander) through a computerized survey. Results indicated that experiencing more positive consequences in the past year was associated with viewing those consequences as both more likely to occur and more positive, while experiencing more negative consequences was associated with viewing them as less negative and no more likely to occur, except for those who had experienced the highest levels of negative consequences. These findings suggest that finding ways to reduce both perceptions as well as consequences themselves may be effective intervention tools. PMID- 21639599 TI - Managing and creating an image in the interview: the role of interviewee initial impressions. AB - In employment interviews, individuals use impression management tactics to present themselves as suitable candidates to interviewers. However, not all impression management tactics, or the interviewees who employ them, are effective at positively influencing interview scores. Results of this study indicate that the relationship between impression management tactic usage and interview success is contingent on the type of tactic employed. Specifically, self-promotion was found to have a positive relationship (r = .20) and slight (r = -.11) and extensive (r = -.19) image creation were found to have negative relationships with end-of-interview scores. Further, the relationships between these 3 impression management tactics were moderated by interviewees' initial impressions of the interview (DeltaR2 ranged from .04 to .10). Interviewees who perceived they were seen as less suitable during rapport building were more effective when using any of the 3 impression management tactics. PMID- 21639600 TI - The relations of daily task accomplishment satisfaction with changes in affect: a multilevel study in nurses. AB - Focusing on a sample of nurses, this investigation examined the relationships of daily task accomplishment satisfaction (for direct and indirect care tasks) with changes in positive and negative affect from preshift to postshift. Not accomplishing tasks to one's satisfaction was conceptualized as a daily workplace stressor that should increase daily negative affect and decrease daily positive affect from preshift to postshift. Further, because of the greater centrality of direct care nursing tasks to nursing work role identities (relative to indirect care tasks), we expected that task accomplishment satisfaction (or lack thereof) for these tasks would have stronger effects on changes in affect than would task accomplishment satisfaction for indirect care tasks. We also examined 2 person level resources, collegial nurse-physician relations and psychological resilience, as moderators of the relationships among these daily variables, with the expectation that these resources would buffer the harmful effects of low task accomplishment satisfaction on nurse affect. Results supported almost all of the proposed effects, though the cross-level interactions were observed only for the effects of indirect care task accomplishment satisfaction on affect and not for direct care task accomplishment satisfaction on affect. PMID- 21639601 TI - The role of context in re-acquisition of extinguished alcoholic beer-seeking. AB - We studied the role of context in reacquisition of extinguished reward-seeking. Rats were trained to respond for alcoholic beer, then extinguished and retrained. Reacquisition was faster than acquisition regardless of whether retraining occurred in the original training context, the extinction context, a novel context, or a context with a mixed history of reinforcement. Reacquisition was also rapid after extended extinction training. Nonetheless, context did significantly influence reacquisition via affecting latency to first response: rats took significantly longer to initiate responding when tested in the extinction context. These results suggest that reacquisition of drug and reward seeking is determined by an inhibitory influence caused by the extinction context and a facilitatory influence caused by reintroduction of the reinforcer (Bouton, 1993). PMID- 21639602 TI - Introduction to the special section on "translational models of prefrontal cortical function". AB - Impaired functioning of the prefrontal cortex is particularly prominent in many forms of psychopathology and in degenerative brain diseases. Because it is challenging to draw causal links between specific brain abnormalities and impaired cognition in these conditions, research using nonhuman animals has a key role to play in elucidating the neurobiological mechanisms of prefrontal cortex function and aiding the search for treatments. This role is clearly illustrated in the review articles and original research reports in this special section. Taken together, these papers demonstrate the insights that have already been gained from research with nonhuman animals as well as the work that still needs to be done to attain the goal of understanding human prefrontal cortical function in both health and disease. PMID- 21639603 TI - Animal models of prefrontal-executive function. AB - Executive function allows us to interact with the world in a purposive, goal directed manner. It relies on several cognitive control operations that are mediated by different regions of the prefrontal cortex. While much of our knowledge about the functional subdivisions of the prefrontal cortex comes from the systematic assessment of patients with brain damage, animal models have served as the predominant tool for investigating specific structure-function relationships within the prefrontal cortex, especially as they relate to complex executive behaviors. These studies generally involve the targeted disruption of neural circuits combined with behavioral testing using carefully designed cognitive paradigms. In this review, I will describe a broad range of such experiments conducted in rats and monkeys that together reveal the distinct contributions of dorsal, medial, and ventral prefrontal cortex to different aspects of executive function. The effects of lesions and local pharmacological manipulations have provided valuable insights into the neural underpinnings of executive function and its neurochemical modulation. Despite the challenges associated with establishing a precise homology between animal models of prefrontal function and the human brain, such models currently offer the best means to systematically investigate the cognitive building blocks of executive function. This helps define the neural circuits that lead to a range of neurological and psychiatric disorders and facilitate the development of effective therapeutic strategies to ameliorate the associated cognitive impairments. PMID- 21639604 TI - Orbitofrontal cortex and basolateral amygdala lesions result in suboptimal and dissociable reward choices on cue-guided effort in rats. AB - The orbitofrontal cortex (OFC) and basolateral nucleus of the amygdala (BLA) are important neural regions in responding adaptively to changes in the incentive value of reward. Recent evidence suggests these structures may be differentially engaged in effort and cue-guided choice behavior. In 2 T-maze experiments, we examined the effects of bilateral lesions of either BLA or OFC on (1) effortful choices in which rats could climb a barrier for a high reward or select a low reward with no effort and (2) effortful choices when a visual cue signaled changes in reward magnitude. In both experiments, BLA rats displayed transient work aversion, choosing the effortless low reward option. OFC rats were work averse only in the no cue conditions, displaying a pattern of attenuated recovery from the cue conditions signaling reward unavailability in the effortful arm. Control measures rule out an inability to discriminate the cue in either lesion group. PMID- 21639605 TI - A biopsychosocial formulation of pain communication. AB - We present a detailed framework for understanding the numerous and complicated interactions among psychological and social determinants of pain through examination of the process of pain communication. The focus is on an improved understanding of immediate dyadic transactions during painful events in the context of broader social phenomena. Fine-grain consideration of social transactions during pain leads to an appreciation of sociobehavioral events affecting both suffering persons as well as caregivers. Our examination considers knowledge from a variety of perspectives, including clinical health psychology, social and developmental processes, evolutionary psychology, communication studies, and behavioral neuroscience. PMID- 21639606 TI - Are leader stereotypes masculine? A meta-analysis of three research paradigms. AB - This meta-analysis examined the extent to which stereotypes of leaders are culturally masculine. The primary studies fit into 1 of 3 paradigms: (a) In Schein's (1973) think manager-think male paradigm, 40 studies with 51 effect sizes compared the similarity of male and leader stereotypes and the similarity of female and leader stereotypes; (b) in Powell and Butterfield's (1979) agency communion paradigm, 22 studies with 47 effect sizes compared stereotypes of leaders' agency and communion; and (c) in Shinar's (1975) masculinity-femininity paradigm, 7 studies with 101 effect sizes represented stereotypes of leadership related occupations on a single masculinity-femininity dimension. Analyses implemented appropriate random and mixed effects models. All 3 paradigms demonstrated overall masculinity of leader stereotypes: (a) In the think manager think male paradigm, intraclass correlation = .25 for the women-leaders similarity and intraclass correlation = .62 for the men-leaders similarity; (b) in the agency-communion paradigm, g = 1.55, indicating greater agency than communion; and (c) in the masculinity-femininity paradigm, g = 0.92, indicating greater masculinity than the androgynous scale midpoint. Subgroup and meta regression analyses indicated that this masculine construal of leadership has decreased over time and was greater for male than female research participants. In addition, stereotypes portrayed leaders as less masculine in educational organizations than in other domains and in moderate- than in high-status leader roles. This article considers the relation of these findings to Eagly and Karau's (2002) role congruity theory, which proposed contextual influences on the incongruity between stereotypes of women and leaders. The implications for prejudice against women leaders are also considered. PMID- 21639607 TI - The dependability of alliance assessments: the alliance-outcome correlation is larger than you might think. AB - OBJECTIVE: To examine the dependability of alliance scores at the patient and therapist level, to evaluate the potential causal direction of session-to-session changes in alliance and depressive symptoms, and to investigate the impact of aggregating the alliance over progressively more sessions on the size of the alliance-outcome relationship. METHOD: We used data from a study (N = 45 patients; N = 9 therapists) of psychotherapy for major depressive disorder in which the alliance was measured at every treatment session to calculate generalizability coefficients and to predict change in depressive symptoms from alliance scores. Two replication samples were also used. RESULTS: At the therapist level, a large number of patients (about 60) per therapist is needed to provide a dependable therapist-level alliance score. At the patient level, generalizability coefficients revealed that a single assessment of the alliance is only marginally acceptable. Very good (>.90) dependability at the patient level is only achieved through aggregating 4 or more assessments of the alliance. Session-to-session change in the alliance predicted subsequent session-to-session changes in symptoms. Evidence for reverse causation was found in later-in treatment sessions, suggesting that only aggregates of early treatment alliance scores should be used to predict outcome. Session 3 alliance scores explained 4.7% of outcome variance, but the average of Sessions 3-9 explained 14.7% of outcome variance. CONCLUSION: Adequate assessment of the alliance using multiple patients per therapist and at least 4 treatment sessions is crucial for fully understanding the size of the alliance-outcome relationship. PMID- 21639608 TI - Nonverbal synchrony in psychotherapy: coordinated body movement reflects relationship quality and outcome. AB - OBJECTIVE: The authors quantified nonverbal synchrony--the coordination of patient's and therapist's movement--in a random sample of same-sex psychotherapy dyads. The authors contrasted nonverbal synchrony in these dyads with a control condition and assessed its association with session-level and overall psychotherapy outcome. METHOD: Using an automated objective video analysis algorithm (Motion Energy Analysis; MEA), the authors calculated nonverbal synchrony in (n = 104) videotaped psychotherapy sessions from 70 Caucasian patients (37 women, 33 men, mean age = 36.5 years, SD = 10.2) treated at an outpatient psychotherapy clinic. The sample was randomly drawn from an archive (N = 301) of routinely videotaped psychotherapies. Patients and their therapists assessed session impact with self-report post-session questionnaires. A battery of pre- and postsymptomatology questionnaires measured therapy effectiveness. RESULTS: The authors found that nonverbal synchrony is higher in genuine interactions contrasted with pseudointeractions (a control condition generated by a specifically designed shuffling procedure). Furthermore, nonverbal synchrony is associated with session-level process as well as therapy outcome: It is increased in sessions rated by patients as manifesting high relationship quality and in patients experiencing high self-efficacy. Higher nonverbal synchrony characterized psychotherapies with higher symptom reduction. CONCLUSIONS: The results suggest that nonverbal synchrony embodies the patients' self-reported quality of the relationship and further variables of therapy process. This hitherto overlooked facet of therapeutic relationships might prove useful as an indicator of therapy progress and outcome. PMID- 21639609 TI - Examining infants' preferences for tempo in lullabies and playsongs. AB - Caregivers around the world sing to their infants. Infants not only prefer to listen to infant-directed singing over adult-directed singing, but infant directed singing also serves a function, communicating affective information to preverbal infants to aid in adjusting arousal levels. Pitch variation has previously been identified as one performance feature that may help to convey the message. Earlier research has indicated that infants' pitch preferences are context dependent, suggesting that infants are tuned in to the communicative intent of infant-directed singing. However, there are several other performance based features present in infant-directed singing that may also contribute to the affective message. The current study examined the role of context on infants' tempo preferences in sung playsongs and lullabies. Using a head-turn preference procedure, we measured 24 preverbal infants' natural preferences for foreign language playsongs and lullabies as a function of tempo. Infants showed a preference for fast over slow tempo playsongs, but no such context dependent preference was found within lullabies. Results partially support the role of tempo as a communicative feature of infant directed singing. PMID- 21639610 TI - Dynamics of the spelling process during a copy task: effects of regularity and frequency. AB - This study investigated the time course of spelling, and its influence on graphomotor execution, in a successive word copy task. According to the cascade model, these two processes may be engaged either sequentially or in parallel, depending on the cognitive demands of spelling. In this experiment, adults were asked to copy a series of words varying in frequency and spelling regularity. A combined analysis of eye and pen movements revealed periods where spelling occurred in parallel with graphomotor execution, but concerned different processing units. The extent of this parallel processing depended on the words' orthographic characteristics. Results also highlighted the specificity of word recognition for copying purposes compared with recognition for reading tasks. The results confirm the validity of the cascade model and clarify the nature of the dependence between spelling and graphomotor processes. PMID- 21639611 TI - Episodic, generalized, and semantic memory tests: switching and strength effects. AB - We continue the process of investigating the probabilistic paired associate paradigm in an effort to understand the memory access control processes involved and to determine whether the memory structure produced is in transition between episodic and semantic memory. In this paradigm two targets are probabilistically paired with a cue across a large number of short lists. Participants can recall the target paired with the cue in the most recent list (list specific test), produce the first of the two targets that have been paired with that cue to come to mind (generalised test), and produce a free association response (semantic test). Switching between a generalised test and a list specific test did not produce a switching cost indicating a general similarity in the control processes involved. In addition, there was evidence for a dissociation between two different strength manipulations (amount of study time and number of cue-target pairings) such that number of pairings influenced the list specific, generalised and the semantic test but amount of study time only influenced the list specific and generalised test. PMID- 21639612 TI - Spatial effects on tactile duration categorization. AB - The aim of this study was to measure the impact of the distance between tactile stimuli marking brief time intervals on perceived duration and threshold estimates. Each interval to be categorized as short or long (midvalue=500 ms) was marked by two brief signals delivered on participants' left (L) or right (R) hand: L-L, R-R, L-R or R-L. The hands were placed nearby or at a distance of 3 feet (about 91 cm). Eight-point individual psychometric functions were drawn for each of the eight experimental conditions. The results reveal that when intervals are marked with signals delivered on different hands rather than on the same hand, they are perceived as longer. Moreover, no difference for perceived duration was observed between the L-L and R-R conditions, and between the L-R and R-L sequence. Finally, marking intervals with signals delivered at the same hand results in better temporal discrimination than with one signal delivered on each hand. The results with perceived duration are consistent with the kappa effect, but not with an attentional account of duration discrimination. PMID- 21639613 TI - Modulating the interference effect in timing with varying stimulus onset asynchrony. AB - The interference effect on time judgments, when subjects are also required to perform a concurrent nontemporal task, is one of the most reliable findings in the time perception literature. In the present study, the interference between a time discrimination task (short or long tone) and a digit classification task (even or odd digit) was analysed using the overlapping tasks paradigm. Reaction times in the digit task were shorter at longer values of stimulus onset asynchrony (SOA) in Experiment 1, showing a clear modulation of interference with varying the relative position of the tasks. Using longer tone durations in Experiment 2, reaction times in the digit task were affected not only by the overlap between the tasks but also by the temporal proximity of responses in the timing and digit tasks. In Experiment 3, the effect of varying the SOA on performance on the digit task was abolished when the auditory tone was irrelevant, thus eliminating an interpretation in terms of distraction from the tone offset. We conclude that the interference effect in concurrent time discrimination and digit classification may be modulated by the degree of overlap between the tasks as well as by the overlap between late processing stages related to decision and response components in the 2 tasks. PMID- 21639614 TI - Mechanisms of what-where-when memory in black-capped chickadees (Poecile atricapillus): do chickadees remember "when"? AB - Integrated what-where-when memory, sometimes called 'episodic-like' memory, has been shown to occur in a number of animals, including food-storing birds and rodents. It is not always clear in these studies, however, what aspect of "when" is remembered. We examined memory for what, where, and when in black-capped chickadees Poecile atricapillus in a procedure designed to dissociate memory for elapsed time (how long ago) from memory for a point in time, the sense of "when" implied in most discussions of episodic or episodic-like memory. Chickadees searched for food in multiple trials that each involved a two-phase procedure. In Phase 1, birds found preferred mealworms in half the sites on one side of an aviary and less preferred sunflower seeds in half the sites on the other side of the aviary. In Phase 2, following a retention interval, birds searched in the aviary with fresh sunflower seeds placed in the sites that held seeds in Phase 1 but either fresh or degraded mealworms in the sites that held mealworms in Phase 1. Whether a site held fresh or degraded worms depended on either when during the day Phase 1 had occurred (Group When), how long ago Phase 1 had occurred (Group HLA), or both (Group When + HLA). Chickadees in all three groups were able to discriminate where and when palatable worms were to be found, but there were temporal limitations on the use of HLA and When + HLA cues that were not found for the use of When cues. PMID- 21639615 TI - "Boys don't cry": examination of the links between endorsement of masculine norms, self-stigma, and help-seeking attitudes for men from diverse backgrounds. AB - The role of conformity to dominant U.S. masculine norms as an antecedent to help seeking attitudes in men has been established using convenience samples made up largely of college-age and European American males. However, the role of conformity to masculine norms on help-seeking attitudes for noncollege-age men or for men from diverse backgrounds is not well understood. To fill this gap in the literature, the present study examined the cross-cultural relevance of a mediational model of the relationships between conformity to dominant U.S. masculine norms and attitudes toward counseling through the mediator of self stigma of seeking counseling for 4,773 men from both majority and nonmajority populations (race/ethnicity and sexual orientation). Structural equation modeling results showed that the model established using college males from majority groups (European American, heterosexual) may be applicable to a community sample of males from differing racial/ethnic groups and sexual orientations. However, some important differences in the presence and strengths of the relationships between conformity to dominant masculine norms and the other variables in the model were present across different racial/ethnic groups and sexual orientations. These findings suggest the need to pay specific theoretical and clinical attention to how conformity to dominant masculine norms and self-stigma are linked to unfavorable attitudes toward help seeking for these men, in order to encourage underserved men's help-seeking behavior. PMID- 21639616 TI - People--things and data--ideas: bipolar dimensions? AB - We examined a longstanding assumption in vocational psychology that people-things and data-ideas are bipolar dimensions. Two minimal criteria for bipolarity were proposed and examined across 3 studies: (a) The correlation between opposite interest types should be negative; (b) after correcting for systematic responding, the correlation should be greater than -.40. In Study 1, a meta analysis using 26 interest inventories with a sample size of 1,008,253 participants showed that meta-analytic correlations between opposite RIASEC (realistic, investigative, artistic, social, enterprising, conventional) types ranged from -.03 to .18 (corrected meta-analytic correlations ranged from -.23 to -.06). In Study 2, structural equation models (SEMs) were fit to the Interest Finder (IF; Wall, Wise, & Baker, 1996) and the Interest Profiler (IP; Rounds, Smith, Hubert, Lewis, & Rivkin, 1999) with sample sizes of 13,939 and 1,061, respectively. The correlations of opposite RIASEC types were positive, ranging from .17 to .53. No corrected correlation met the criterion of -.40 except for investigative-enterprising (r = -.67). Nevertheless, a direct estimate of the correlation between data-ideas end poles using targeted factor rotation did not reveal bipolarity. Furthermore, bipolar SEMs fit substantially worse than a multiple-factor representation of vocational interests. In Study 3, a two-way clustering solution on IF and IP respondents and items revealed a substantial number of individuals with interests in both people and things. We discuss key theoretical, methodological, and practical implications such as the structure of vocational interests, interpretation and scoring of interest measures for career counseling, and expert RIASEC ratings of occupations. PMID- 21639617 TI - Imitation in young children: when who gets copied is more important than what gets copied. AB - Unlike other animals, human children will copy all of an adult's goal-directed actions, including ones that are clearly unnecessary for achieving the demonstrated goal. Here we highlight how social affiliation is key to this species-specific behavior. Preschoolers watched 2 adults retrieve a toy from a novel apparatus. One adult included irrelevant actions in her demonstration; the other only used actions causally related to opening the apparatus. After both adults took turns demonstrating, 1 left the test room, and the remaining adult gave the apparatus to the child. Children reproduced the irrelevant actions only when given the apparatus by the adult who had demonstrated them, even though the departed adult's actions emphasized how unnecessary these redundant actions were. Our results highlight the specialized skills for participating in cultural groups that have evolved in humans and provide insight into why finding such high fidelity copying in other animals has proven elusive. PMID- 21639618 TI - The contribution of extracurricular activities to adolescent friendships: new insights through social network analysis. AB - Extracurricular activities are settings that are theorized to help adolescents maintain existing friendships and develop new friendships. The overarching goal of the current investigation was to examine whether coparticipating in school based extracurricular activities supported adolescents' school-based friendships. We used social network methods and data from the National Longitudinal Study of Adolescent Health to examine whether dyadic friendship ties were more likely to exist among activity coparticipants while controlling for alternative friendship processes, namely dyadic homophily (e.g., demographic and behavioral similarities) and network-level processes (e.g., triadic closure). Results provide strong evidence that activities were associated with current friendships and promoted the formation of new friendships. These associations varied based on school level (i.e., middle vs. high school) and activity type (i.e., sports, academic, arts). Results of this study provide new insight into the complex relations between activities and friendship that can inform theories of their developmental outcomes. PMID- 21639620 TI - Culture and the sequence of steps in theory of mind development. AB - To examine cultural contrasts in the ordered sequence of conceptual developments leading to theory of mind (ToM), we compared 135 3- to 6-year-olds (77 Australians; 58 Iranians) on an established 5-step ToM scale (Wellman & Liu, 2004). There was a cross-cultural difference in the sequencing of ToM steps but not in overall rates of ToM mastery. In line with our predictions, the children from Iran conformed to a distinctive sequence previously observed only in children in China. In contrast to the case with children from Australia (and the United States), knowledge access was understood earlier than opinion diversity in children from Iran, consistent with this collectivist culture's emphasis on filial respect, dispute avoidance, and acquiring knowledge. Having a sibling was linked with faster overall ToM progress in Australia only and was not related to scale sequences in either culture. PMID- 21639619 TI - Developmental changes in anger expression and attention focus: learning to wait. AB - Being able to wait is an essential part of self-regulation. In the present study, the authors examined the developmental course of changes in the latency to and duration of target-waiting behaviors by following 65 boys and 55 girls from rural and semirural economically strained homes from ages 18 months to 48 months. Age related changes in latency to and duration of children's anger expressions and attention focus (e.g., self-initiated distraction) during an 8-min wait for a gift were found. On average, at 18 and 24 months of age, children were quick to react angrily and slower to shift attention away from the desired object than they were at later ages. Over time, children were quicker to distract themselves. By 36 months, distractions occurred before children expressed anger, and anger expressions were briefer. At 48 months, children typically made a quick bid to their mothers about having to wait before distracting themselves; on average, they did not appear angry until the latter half of the wait. Unexpectedly, children bid to their mothers as much at age 48 months as they had at 18 months; however, bids became less angry as children got older. Developmental changes in distraction and bidding predicted age-related changes in the latency to anger. Findings are discussed in terms of the neurocognitive control of attention around age 30 months, the limitations of children's self-regulatory efforts at age 48 months, and the importance of fostering children's ability to forestall, as well as modulate, anger. PMID- 21639621 TI - Similarity in depressive symptoms in adolescents' friendship dyads: selection or socialization? AB - This study examined friendship selection and socialization as mechanisms explaining similarity in depressive symptoms in adolescent same-gender best friend dyads. The sample consisted of 1,752 adolescents (51% male) ages 12-16 years (M = 13.77, SD = 0.73) forming 487 friend dyads and 389 nonfriend dyads (the nonfriend dyads served as a comparison group). To test our hypothesis, we applied a multigroup actor-partner interdependence model to 3 friendship types that started and ended at different time points during the 2 waves of data collection. Results showed that adolescents reported levels of depressive symptoms at follow-up that were similar to those of their best friends. Socialization processes explained the increase in similarity exclusively in female dyads, whereas no evidence for friendship selection emerged for either male or female dyads. Additional analyses revealed that similarity between friends was particularly evident in the actual best friend dyads (i.e., true best friends), in which evidence for socialization processes emerged for both female and male friend dyads. Findings highlight the importance of examining friendship relations as a potential context for the development of depressive symptoms. PMID- 21639622 TI - The role of task persistence in young adolescence for successful educational and occupational attainment in middle adulthood. AB - The main purpose of the present study was to investigate the importance of task persistence in young adolescence for successful educational and occupational attainment in middle adulthood. Data from age 13 (N = 1,092) and adult age (age 43 for women, N = 569 and age 47 for men, N = 393) were taken from the Swedish longitudinal research program entitled "Individual Development and Adaptation." In line with previous research, task persistence was found to be related to changes in grades between age 13 and age 16, over and above other childhood factors. Task persistence at age 13 was also a significant predictor of both income and occupational level in middle adulthood for the men, controlling for a number of childhood factors (including intelligence), and even when educational attainment in adulthood was taken into account. Finally, task persistence was related to educational attainment in adulthood. The authors suggest that task persistence is a second fundamental factor besides general mental ability, influencing attainment within the area of working life and education. The influence of task persistence is discussed in form of personality-environment selection mechanisms. PMID- 21639623 TI - Individual differences in boys' and girls' timing and tempo of puberty: modeling development with nonlinear growth models. AB - Pubertal development is a nonlinear process progressing from prepubescent beginnings through biological, physical, and psychological changes to full sexual maturity. To tether theoretical concepts of puberty with sophisticated longitudinal, analytical models capable of articulating pubertal development more accurately, we used nonlinear mixed-effects models to describe both the timing and tempo of pubertal development in the sample of 364 White boys and 373 White girls measured across 6 years as part of the National Institute of Child Health and Human Development Study of Early Child Care and Youth Development. Individual differences in timing and tempo were extracted with models of logistic growth. Differential relations emerged for how boys' and girls' timing and tempo of development were related to physical characteristics (body mass index, height, and weight) and psychological outcomes (internalizing problems, externalizing problems, and risky sexual behavior). Timing and tempo are associated in boys but not girls. Pubertal timing and tempo are particularly important for predicting psychological outcomes in girls but only sparsely related to boys' psychological outcomes. Results highlight the importance of considering the nonlinear nature of puberty and expand the repertoire of possibilities for examining important aspects of how and when pubertal processes contribute to development. PMID- 21639624 TI - Maternal depressive symptomatology and child behavior: transactional relationship with simultaneous bidirectional coupling. AB - The present study investigated reciprocal relationships between adolescent mothers and their children's well-being through an analysis of the coupling relationship of mothers' depressive symptomatology and children's internalizing and externalizing behaviors. Unlike studies using discrete time analyses, the present study used dynamical systems to model time continuously, which allowed for the study of dynamic, transactional effects between members of each dyad. Findings provided evidence of coupling between maternal depressive symptoms and children's behaviors. The most robust finding was that as maternal depressive symptoms became more or less severe, children's behavior problems increased or decreased in a reciprocal manner. Results from this study extended upon theoretical contributions of such authors as Richters (1997) and Granic and Hollenstein (2003), providing empirical validation from a longitudinal study for understanding the ongoing, dynamic relationships between at-risk mothers and their children. PMID- 21639625 TI - Antagonistic pleiotropy at the human IL6 promoter confers genetic resilience to the pro-inflammatory effects of adverse social conditions in adolescence. AB - The authors tested the evolutionary genetic hypothesis that the functional form of an asymmetrically risky Gene * Environment interaction will differ as a function of age-related antagonistic pleiotropy (i.e., show opposite effects in young vs. old individuals). Previous studies have identified a polymorphism in the human IL6 promoter (rs1800795; IL6-74 G/C) that interacts with adverse socioenvironmental conditions to promote chronic inflammation in older adults (elevated C-reactive protein). This study identifies a protective effect of the same polymorphism in 17- to 19-year-old adolescents confronting socioeconomic adversity. Over 60% of the environmental risk contribution to the IL6 * Socioeconomic Status interaction could be accounted for by interpersonal stress and adult role burden. Thus, the IL6-174G allele does not represent an undifferentiated risk factor but instead sensitizes inflammatory biology to socioenvironmental conditions, conferring either genetic vulnerability or resilience depending on the developmental "somatic environment" that interacts with social conditions to influence gene expression. PMID- 21639626 TI - Personality types in adolescence: change and stability and links with adjustment and relationships: a five-wave longitudinal study. AB - We examined change and stability of the 3 personality types identified by Block and Block (1980) and studied their links with adjustment and relationships. We used data from a 5-wave study of 923 early-to-middle and 390 middle-to-late adolescents, thereby covering the ages of 12-20 years. In Study 1, systematic evidence for personality change was found, in that the number of overcontrollers and undercontrollers decreased, whereas the number of resilients increased. Undercontrol, in particular, was found to peak in early-to-middle adolescence. We also found substantial stability of personality types, because 73.5% of the adolescents had the same personality type across the 5 waves. Personality change was mainly characterized by 2 transitions: overcontrol -> resiliency and undercontrol -> resiliency. The transitional analyses implied that the resilient type serves more often as the end point of personality development in adolescence than do overcontrol and undercontrol. Analyses of the personality type trajectories also revealed that the majority of adolescents who change personality type across 5 years made only 1 transition. Study 2 revealed systematic differences between resilients and overcontrollers in anxiety. Stable resilients were less anxious over time than were stable overcontrollers. Further, change from overcontrol to the resilient type was accompanied by decreases in anxiety, whereas change from the resilient type to overcontrol was accompanied by an increase in anxiety. Similarly, systematic differences between personality types were found in the formation of intimate relationships. PMID- 21639627 TI - Don't sit so close to me: unconsciously elicited affect automatically provokes social avoidance. AB - Behavior may be automatically prompted by cues in our social environment. Previous research has focused on cognitive explanations for such effects. Here we hypothesize that affective processes are susceptible to similar automatic influences. We propose that exposure to groups stereotyped as dangerous or violent may provoke an anxiety response and, thus, a tendency to move away. In the present experiment, we subliminally exposed participants to images of such a group, and found that they displayed greater avoidance in a subsequent interaction. Critically, this effect was explained by their increased sensitivity to threat-related information. These findings demonstrate an affective mechanism responsible for nonconscious priming effects on interpersonal behavior. PMID- 21639628 TI - Should I go with my gut? Investigating the benefits of emotion-focused decision making. AB - Deliberative decision strategies have historically been considered the surest path to sound decisions; however, recent evidence and theory suggest that affective strategies may be equally as effective. In four experiments we examined conditions under which affective versus deliberative decision strategies might result in higher decision quality. While consciously focusing on feelings versus details, participants made choices that varied in complexity, in extent of subsequent conscious deliberation allowed, and in domain. Results indicate that focusing on feelings versus details led to superior objective and subjective decision quality for complex decisions. However, when using a feeling-focused approach, subsequent deliberation after encoding resulted in reduced choice quality. These results suggest that affective decision strategies may be more effective relative to deliberative strategies for certain complex decisions. PMID- 21639629 TI - Misery loves company: mood-congruent emotional responding to music. AB - We examined emotional responding to music after mood induction. On each trial, listeners heard a 30-s music excerpt and rated how much they liked it, whether it sounded happy or sad, and how familiar it was. When the excerpts sounded unambiguously happy or sad (Experiment 1), the typical preference for happy sounding music was eliminated after inducing a sad mood. When the excerpts sounded ambiguous with respect to happiness and sadness (Experiment 2), listeners perceived more sadness after inducing a sad mood. Sad moods had no influence on familiarity ratings (Experiments 1 and 2). These findings imply that "misery loves company." Listeners in a sad mood fail to show the typical preference for happy-sounding music, and they perceive more sadness in music that is ambiguous with respect to mood. PMID- 21639630 TI - Depressive symptoms and mechanisms of relational turbulence as predictors of relationship satisfaction among returning service members. AB - In contrast to romanticized portrayals of reunion after deployment, U.S. military personnel may contend with the harsh reality of both depressive symptoms and upheaval in their romantic relationships during the postdeployment transition. This study employed the relational turbulence model to evaluate mechanisms linking depressive symptoms with relationship satisfaction. Cross-sectional, self report data were collected from 220 service members living in 27 states who had returned home from deployment within the past six months. As hypothesized, the negative association between depressive symptoms and relationship satisfaction was mediated by relational uncertainty and interference from partners. These findings advance scholarship on depressive symptoms and relational turbulence, and they also suggest guidelines for helping service members with depressive symptoms maintain satisfying romantic relationships upon reentry. PMID- 21639631 TI - Long-term impact of parental divorce on intimate relationship quality in adulthood and the mediating role of psychosocial resources. AB - The purpose of this 16-year prospective follow-up study was to investigate the association between parental divorce in childhood and intimate relationship quality in adulthood. The mediating role of psychosocial resources (parent-child relationships at 16 years, self-esteem and social support at 32 years) in this association was also studied. All 16 year olds of one Finnish city completed questionnaires at school and were followed up by postal questionnaires at 32 years of age (n = 1,471). Results showed that women and men from divorced families were more often divorced or separated at the age of 32 years than those from nondivorced families. However, parental divorce was associated with poorer intimate relationship quality only among women. Women from divorced families also had poorer relationships with their father and mother in adolescence, and they had lower self-esteem and satisfaction with social support in adulthood than women from intact families. No such associations were found among men. The impact of parental divorce on intimate relationship quality among women was partially mediated by mother-daughter relationship, self-esteem, and satisfaction with social support. The mediating role of mother-daughter relationship was not direct, however, but was mediated via self-esteem and satisfaction with social support. Our findings indicate that parental divorce affects daughters more than sons. In the context of parental divorce, the mother-daughter relationship in adolescence is important for the development of later psychosocial resources and, via them, for intimate relationship quality. PMID- 21639632 TI - The language of demand/withdraw: verbal and vocal expression in dyadic interactions. AB - Associations between vocally expressed emotional arousal, influence tactics, and demand/withdraw behavior were examined in a treatment-seeking sample of 130 seriously and stably distressed, married, heterosexual couples and in a community sample (N = 38) of 18 married heterosexual and 20 dating heterosexual couples. Fundamental frequency was used to measure emotional arousal, and computational linguistics were used to measure influence tactics. Higher levels of demand/withdraw behavior were associated with greater use of manipulative and controlling influence tactics, higher levels of emotional arousal, and less frequent use of cooperative and compromising influence tactics. Overall, demanders tended to express more arousal and to use more influence tactics than withdrawers. Both influence tactics and emotional arousal were uniquely associated with demand/withdraw behavior. Implications of results are discussed for refining theories of demand/withdraw interaction. PMID- 21639633 TI - Couple adjustment and posttraumatic stress disorder symptoms in National Guard veterans of the Iraq war. AB - Relationship adjustment and posttraumatic stress disorder (PTSD) symptoms were assessed across two time points in a sample of 313 married or partnered National Guard soldiers recently returned from combat duty in Iraq. Structural equation modeling using a four-factor model for PTSD found the latent variable dysphoria (reflecting generalized distress including aspects of emotional numbing and arousal) had the strongest independent contribution to predicting relationship adjustment at Time 1 and indirectly predicted poorer relationship adjustment at Time 2. Exploratory analysis of gender differences (n = 33 women; n = 280 men) suggested a different pattern of relations between PTSD factors and relationship adjustment among female soldiers at Time 1, with a trend toward trauma specific avoidance being more highly related to relationship adjustment. Clinical and research implications are discussed. PMID- 21639634 TI - The implication of combat-induced stress reaction, PTSD, and attachment in parenting among war veterans. AB - This study examined parental functioning, parental satisfaction, and concern for offspring during their child's military service, among war veterans, some of whom suffered from acute combat-induced stress reaction (CSR) and posttraumatic stress disorder (PTSD). In addition, we examined the additive and interactive contributions of CSR, PTSD and attachment dimensions to parenting measures. The sample consisted of 477 participants divided into two groups: a clinical group of veterans who had been diagnosed with CSR on the battlefield (N = 267), and a matched control group of veterans who did not suffer from CSR (NCSR; N = 210). CSR, PTSD, avoidant-attachment, and anxious-attachment, were all related to lower levels of parental functioning and satisfaction. Veterans who suffered from both CSR and PTSD reported more concern for their offspring during their child's military service compared to veterans with PTSD but without antecedent CSR. Attachment dimensions and specifically attachment-avoidance, made the greatest contribution to parenting measures, followed by posttraumatic symptoms. In addition, attachment-avoidance moderated the relationship between posttraumatic symptoms and parental functioning. Theoretical and clinical implications of these results are discussed. PMID- 21639635 TI - Distress in spouses of service members with symptoms of combat-related PTSD: secondary traumatic stress or general psychological distress? AB - Combat-related posttraumatic stress disorder (PTSD) is linked with elevated psychological distress in service members'/veterans' spouses. Researchers use a variety of terms to describe this distress, and recently, secondary traumatic stress and secondary traumatic stress disorder (STS/STSD) have become increasingly commonly used. Although STS/STSD connotes a specific set of symptoms that are linked to service members'/veterans' symptoms, researchers often use general measures of distress or generically worded measures of PTSD symptoms to assess STS/STSD. To determine how often scores on such measures appear to be an accurate reflection of STS/STSD, we examined responses to a measure of PTSD symptoms in 190 wives of male service members with elevated levels of PTSD symptoms. Wives rated their own PTSD symptoms, and then answered questions about their attributions for the symptoms they endorsed. Fewer than 20% of wives who endorsed symptoms on the PTSD measure attributed these symptoms completely to their husbands' military experiences. Moreover, compared with wives who attributed symptoms only to events in their own lives, wives who attributed symptoms completely or partially to their husbands' military experiences had a greater overlap between some of their responses on the PTSD measure and their responses to a measure of general psychological distress. These results suggest that most wives of service members/veterans with PTSD experience generic psychological distress that is not conceptually consistent with STS/STSD, although a subset does appear to endorse a reaction consistent with this construct. Implications of these findings for intervention and research with this vulnerable population are discussed. PMID- 21639636 TI - Preliminary results for an adaptive family treatment for drug abuse in Hispanic youth. AB - A small randomized trial investigated a new family-based intervention for Hispanic adolescents who met DSM-IV criteria for substance abuse disorder. The Culturally Informed and Flexible Family-Based Treatment for Adolescents (CIFFTA) is a tailored/adaptive intervention that includes a flexible treatment manual and multiple treatment components. The study used an "add on" design to isolate the effects on substance abuse, behavior problems, and parenting practices attributable to the newly developed components. Twenty-eight Hispanic adolescents and their families were randomized either to the experimental treatment or to traditional family therapy (TFT) and were assessed at baseline and 8-month follow up. Despite the small sample, results revealed statistically significant time * treatment effects on both self-reported drug use (marijuana + cocaine), F(1, 22) = 10.59, p < .01, eta2 = .33 and adolescent reports of parenting practices, F(1, 22) = 9.01, p < .01, eta2 = .29. Both sets of analyses favored CIFFTA participants. There was a significant time * treatment effect, F(1, 22) = 6.72, p = .02, eta2 = .23, favoring CIFFTA on parent report of parenting practices using a composite that matched the variables used for adolescents, but only a nonsignificant trend, F(1, 22) = 2.43, p = .13, eta2 = .10, with a composite that used all parenting subscales. Parent reports of adolescent behavior problems did not show a significant time or time * treatment effect. These results show the promise of this adaptive treatment for substance abuse in Hispanic adolescents and suggest the need for a larger randomized trial to fully investigate this treatment. PMID- 21639637 TI - Socioeconomic inequalities in colorectal cancer screening uptake: does time perspective play a role? AB - OBJECTIVE: This study examined the role of time perspective in explaining inequalities in colorectal cancer screening attendance. We tested a path model predicting that (a) socioeconomic status (SES) would be associated with consideration of future consequences (CFC), (b) CFC would be associated with perceived benefits/barriers, and (c) barriers and benefits would be associated longitudinally with screening attendance. METHOD: Data for these analyses came from the control arm (n = 809) of an intervention to increase screening uptake. Participants between 55 and 64 years were offered screening as part of the U.K. Flexible Sigmoidoscopy (FS) Trial. They completed a questionnaire that included demographic and psychological variables. Subsequent screening attendance was recorded. RESULTS: There was clear evidence of SES differences in attendance, with 56% in the most deprived tertile attending their FS appointment, compared with 68% in the middle tertile and 71% in the least deprived tertile (p < .01). Lower SES was associated with lower CFC, higher perceived barriers, and lower perceived benefits (p < .05 for all). Higher CFC, higher perceived benefits, and lower perceived barriers were associated with attendance (p < .01 for all). CFC mediated the association between SES and perceived benefits/barriers, while perceived benefits/barriers mediated the association between CFC and attendance. CONCLUSION: SES differences in CFC contribute to SES differences in the perceived barriers and benefits of screening, which, in turn, contribute to differences in attendance. Interventions that take CFC into account, for example, by emphasizing short-term benefits, could promote equality in screening participation. PMID- 21639638 TI - The long-term efficacy of two computer-tailored physical activity interventions for older adults: main effects and mediators. AB - OBJECTIVE: Low-cost (e.g., computer-tailored) interventions with sustained effects are needed to increase and maintain physical activity in older adults. This study examined the long-term efficacy of 2 computer-tailored physical activity interventions for older adults and its psychosocial and environmental mediators. METHODS: A clustered randomized controlled trial (N = 1,971) was conducted that included 3 research arms: (a) basic computer-tailored print intervention, targeting psychosocial mediators; (b) environmentally computer tailored print intervention, targeting psychosocial and environmental mediators; and (c) no-intervention control group. Interventions were developed using the intervention mapping approach and consisted of 3 computer-tailored letters delivered over 4 months. Questionnaires assessed the study outcomes (i.e., total weekly days and total weekly minutes of physical activity) at baseline and 12 months. Potential mediators (i.e., awareness, attitude, self-efficacy, intention, social influence, intrinsic motivation, self-regulation, and perceived environment) were assessed at baseline and at 3 or 6 months. RESULTS: Multilevel regression analyses revealed that both interventions significantly changed total weekly days of physical activity compared with the control group, but only the environmentally computer-tailored print intervention significantly changed weekly minutes of physical activity. Multiple mediation models showed that the effects of both interventions on weekly days of physical activity were mediated by changes in awareness and intention. CONCLUSIONS: Computer-tailored interventions were effective in inducing long-term behavioral changes in physical activity behavior of older adults. Awareness and intention were found to be important mediators of changing daily physical activity and should be included in future computer-tailored intervention studies. PMID- 21639639 TI - Improving everyday error detection, one picture at a time: a performance-based study of everyday task training. AB - OBJECTIVE: Research suggests that dementia patients detect fewer action errors than age-matched controls; however, little is known about the derivation of their error-monitoring difficulties. The aims of the study are to evaluate a novel, task-training action intervention (TT-NAT) designed to increase error monitoring in dementia patients and to pinpoint the relation between error monitoring and neuropsychological processes. METHOD: Participants (n = 45) with dementia were administered the Standard NAT, a performance-based test requiring completion of three everyday tasks. A second group (n = 42) was administered the TT-NAT, which includes a brief training session prior to the commencement of each task. All participants were compared on the following variables: total errors, proportion of errors detected, and proportion of errors corrected. Correlations between error-monitoring variables and neuropsychological tests of executive functioning and language were performed. RESULTS: TT-NAT participants produced fewer total errors and detected significantly more errors than Standard NAT participants (z = 3.0; t = 3.36; p < .05). Error detection was strongly related to only the language composite index (r = .57, p = .00) in the TT-NAT, whereas it was moderately related to both the language (r = .31, p = .04) and executive composite (r = .36, p = .02) indices in the Standard NAT condition. CONCLUSION: Review of task steps and objects before task performance may be a promising intervention for error-monitoring deficits in dementia patients; this finding has implications for neuropsychological rehabilitation of functional deficits in this population. PMID- 21639640 TI - Neonatal respiratory dysfunction and neuropsychological performance at the preschool age: a study of very preterm infants with bronchopulmonary dysplasia. AB - OBJECTIVE: The neuropsychological outcome of chronic lung disease (CLD) in the very preterm (VP) infant may be determined by a process involving chronic hypoxia, with superimposed acute hypoxic episodes, in the developing brain. We wished to study the differences in quality of outcome between VP preschoolers with and without history of the most common form of CLD in the preterm infant, bronchopulmonary dysplasia (BPD). We also examined the strength of association between BPD severity and neuropsychological outcome, with degree of severity defined according to the National Institute of Child Health and Human Development (NICHD) National Heart, Lung and Blood Institute (NHLBI) Workshop categorical ranking scheme (Jobe & Bancalari, 2001) or in accord with dimensional views of severity of respiratory illness. METHOD: We evaluated the intellectual, language, and motor outcomes of 156, predominantly middle-class preschoolers with history of VP birth, with (n = 80) or without (n = 76) BPD. We used supplemental oxygen requirement or need for mechanical ventilation as indirect indexes of respiratory dysfunction. RESULTS: Following adjustment for potentially confounding sociodemographic variables and perinatal medical risk factors, we found no group differences in neuropsychological outcome based on categorical ranking of BPD severity. However, continuous measures of BPD severity accounted for a unique portion of the variance in fine motor performance (eta2p = .05), while patent ductus arteriosus, a risk marker or antecedent of BPD, explained a unique portion of the variance in both receptive language (eta2p = .048), and gross motor (eta2p = .061) function. CONCLUSION: A significant, yet circumscribed, association was demonstrated between neonatal hypoxic risk, in the VP infant, and neuropsychological outcome assessed in the preschool years. PMID- 21639641 TI - Unilateral spatial neglect in degenerative brain pathology. AB - OBJECTIVE: In an attempt to interpret neglect as a disconnection syndrome, it is currently proposed that the disorder results from disorganization of large-scale networks involved in attentional spatial processes rather than of individual brain areas. We hypothesize that as degenerative brain diseases are "system pathologies," degeneration could be restricted to some of the neural subsystems implicated in the functional organization of spatial attention and different neglect syndromes could emerge depending on the patterns of the subsystems involved. METHOD: We studied five neglect patients: one with corticobasal degeneration (CBD), three with Posterior Cortical Atrophy (PCA) and one with frontotemporal dementia (FTD). RESULTS: The patient with CBD and left parietoccipital atrophy showed right allocentric neglect; the three patients with PCA mostly distributed in the right posterior regions showed left egocentric extrapersonal neglect; the patient with FTD, who displayed more severe frontotemporal atrophy on the right, had left motor-executive neglect for both personal and extrapersonal space. All patients also presented a deep breakdown of spatial working memory. CONCLUSION: Our data would confirm that left neglect is more frequent than right neglect also in degenerative pathology and that damage to different neural substrates can produce different types of neglect. Our findings are also consistent with the hypothesis that both lateralized and nonlateralized attention disorders contribute to generate the syndrome. We suggest that evidence from degenerative diseases may contribute to construction of models of spatial attention. PMID- 21639642 TI - On the confounds among retest gains and age-cohort differences in the estimation of within-person change in longitudinal studies: a simulation study. AB - Although longitudinal designs are the only way in which age changes can be directly observed, a recurrent criticism involves to what extent retest effects may downwardly bias estimates of true age-related cognitive change. Considerable attention has been given to the problem of retest effects within mixed effects models that include separate parameters for longitudinal change over time (usually specified as a function of age) and for the impact of retest (specified as a function of number of exposures). Because time (i.e., intervals between assessment) and number of exposures are highly correlated (and are perfectly correlated in equal interval designs) in most longitudinal studies, the separation of effects of within-person change from effects of retest gains is only possible given certain assumptions (e.g., age convergence). To the extent that cross-sectional and longitudinal effects of age differ, obtained estimates of aging and retest may not be informative. The current simulation study investigated the recovery of within-person change (i.e., aging) and retest effects from repeated cognitive testing as a function of number of waves, age range at baseline, and size and direction of age-cohort differences on the intercept and age slope in age-based models of change. Significant bias and Type I error rates in the estimated effects of retest were observed when these convergence assumptions were not met. These simulation results suggest that retest effects may not be distinguishable from effects of aging-related change and age-cohort differences in typical long-term traditional longitudinal designs. PMID- 21639643 TI - Attention and the acquisition of new knowledge: their effects on older adults' associative memory deficit. AB - Older adults experience a selective associative memory deficit by demonstrating intact item memory relative to impaired associative memory when compared with younger adults. Age-related deficits in associative memory have been suggested to arise from declines in attentional resources, and the role of attention during encoding and retrieval in associative memory for words and their spatial locations was investigated in the current experiment. Additionally, the tendency of younger and older adults to use knowledge acquired during encoding to improve their associative memory judgments through a strategic associative memory process was also investigated. Younger and older adults studied a list of words with each word belonging to one of four categories, which followed one of four mathematical probability structures for their presentation. Older adults exhibited intact item memory and impaired associative memory relative to full attention younger adults. In addition, both older and younger adults demonstrated an ability to engage in strategic associative memory, by learning and later using the probability structure introduced at study to guide their associative memory judgments. In contrast, dividing the attention of younger adults during encoding impaired item memory, associative memory and strategic associative memory, whereas dividing attention at retrieval did not result in similar deficits. These data add to a growing body of literature demonstrating older adults to exhibit a selective associative memory deficit that is not simulated by dividing the attention of younger adults at encoding or retrieval. Furthermore, younger and older adults maintain the ability to use new knowledge to guide their associative judgments. PMID- 21639644 TI - Integrative and semantic relations equally alleviate age-related associative memory deficits. AB - Two experiments compared effects of integrative and semantic relations between pairs of words on lexical and memory processes in old age. Integrative relations occur when two dissimilar and unassociated words are linked together to form a coherent phrase (e.g., horse-doctor). In Experiment 1, older adults completed a lexical-decision task where prime and target words were related either integratively or semantically. The two types of relation both facilitated responses compared to a baseline condition, demonstrating that priming can occur in older adults with minimal preexisting associations between primes and targets. In Experiment 2, young and older adults completed a cued recall task with integrative, semantic, and unrelated word pairs. Both integrative and semantic pairs showed significantly smaller age differences in associative memory compared to unrelated pairs. Integrative relations facilitated older adults' memory to a similar extent as semantic relations despite having few preexisting associations in memory. Integratability of stimuli is therefore a new factor that reduces associative deficits in older adults, most likely by supporting encoding and retrieval mechanisms. PMID- 21639645 TI - Genetic and environmental influences on odor identification ability in the very old. AB - Odor identification ability and cognition were measured in a population-based cohort of 1,222 very old twins and singletons, including 91 centenarians. Heritability for identifying odors was low, in contrast to that for cognition. Common genes were found to contribute to both olfaction and cognition. In a multiple regression model, sex, age, cognitive function, and smoking, but not APOEepsilon4 status, were significant predictors of the olfactory test scores (all ps < 0.001). This study, along with data from other studies, suggests that indices of heritability for odor identification decline with age, likely reflecting adverse environmental influences on the smell system. PMID- 21639647 TI - A Combi-Stroop test for measuring food-related attentional bias. AB - There are methodological complexities with the supraliminal-lexical versions of the modified versions of the Stroop tests that could be responsible for inconsistencies across the literature (Field & Cox, 2008). We tested whether a combination of subliminal-pictorial and classic Stroop tests can differentiate between dieters' and nondieters' food attentional bias (FAB). Participants were dieters (n = 30) and nondieters (n = 32) who were tested 3 hr after having a meal. Each picture from among 24 high-calorie and 24 low-calorie food pictures was presented for 32 ms before the appearance of a congruent or an incongruent color word, in response to which participants were required to manually report, via a tagged keyboard, the correct color of the word as quickly and accurately as possible. Color-naming latencies and interference scores were calculated. Dieters showed the highest reaction times to incongruent color words following high calorie food pictures; overall, dieters showed significantly higher FABs than nondieters. The Combi-Stroop test has differential validity. Moreover, findings suggest that FAB can result from early allocation of dieters' attention to food related stimuli. PMID- 21639646 TI - The effects of age on the strategic use of pitch accents in memory for discourse: a processing-resource account. AB - In two experiments, we investigated age-related changes in how prosodic pitch accents affect memory. Participants listened to recorded discourses that contained two contrasts between pairs of items (e.g., one story contrasted British scientists with French scientists and Malaysia with Indonesia). The end of each discourse referred to one item from each pair; these references received a pitch accent that either denoted contrast (L + H* in the ToBI system) or did not (H*). A contrastive accent on a particular pair improved later recognition memory equally for young and older adults. However, older adults showed decreased memory if the other pair received a contrastive accent (Experiment 1). Young adults with low working memory performance also showed this penalty (Experiment 2). These results suggest that pitch accents guide processing resources to important information for both older and younger adults but diminish memory for less important information in groups with reduced resources, including older adults. PMID- 21639648 TI - Affirmation, acknowledgment of in-group responsibility, group-based guilt, and support for reparative measures. AB - Three studies, 2 conducted in Israel and 1 conducted in Bosnia and Herzegovina, demonstrated that affirming a positive aspect of the self can increase one's willingness to acknowledge in-group responsibility for wrongdoing against others, express feelings of group-based guilt, and consequently provide greater support for reparation policies. By contrast, affirming one's group, although similarly boosting feelings of pride, failed to increase willingness to acknowledge and redress in-group wrongdoing. Studies 2 and 3 demonstrated the mediating role of group-based guilt. That is, increased acknowledgment of in-group responsibility for out-group victimization produced increased feelings of guilt, which in turn increased support for reparation policies to the victimized group. Theoretical and applied implications are discussed. PMID- 21639649 TI - Understanding emotional transitions: the interpersonal consequences of changing emotions in negotiations. AB - Research on the interpersonal functions of emotions has focused primarily on steady-state emotion rather than on emotional transitions, the movement between emotion states. The authors examined the influence of emotional transitions on social interactions and found that emotional transitions led to consistently different outcomes than their corresponding steady-state emotions. Across 2 computer-mediated negotiations and a face-to-face negotiation, participants negotiating with partners who displayed a "becoming angry" (happy to angry) emotional transition accepted worse negotiation outcomes yet formed better relational impressions of their partners than participants negotiating with partners who displayed steady-state anger. This relationship was mediated through 2 mechanisms: attributional and emotional contagion processes. The "becoming happy" (angry to happy) emotional transition as compared with steady-state happiness was not significantly related to differences in negotiation outcomes but was significantly related to differences in relational impressions, where perceivers of the "becoming happy" emotional transition gave their partners lower relational impression ratings than perceivers of steady-state happiness. PMID- 21639650 TI - The cognitive consequences of envy: attention, memory, and self-regulatory depletion. AB - In a series of 4 experiments, we provide evidence that--in addition to having an affective component--envy may also have important consequences for cognitive processing. Our first experiment (N = 69) demonstrated that individuals primed with envy better attended to and more accurately recalled information about fictitious peers than did a control group. Studies 2 (N = 187) and 3 (N = 65) conceptually replicated these results, demonstrating that envy elicited by targets predicts attention and later memory for information about them. We demonstrate that these effects cannot be accounted for by admiration or changes in negative affect or arousal elicited by the targets. Study 4 (N = 152) provides evidence that greater memory for envied--but not neutral--targets leads to diminished perseverance on a difficult anagram task. Findings demonstrate that envy may play an important role in attention and memory systems and deplete limited self-regulatory resources available for acts of volition. PMID- 21639651 TI - The time for doing is not the time for change: effects of general action and inaction goals on attitude retrieval and attitude change. AB - Implicit in many informal and formal principles of psychological change is the understudied assumption that change requires either an active approach or an inactive approach. This issue was systematically investigated by comparing the effects of general action goals and general inaction goals on attitude change. As prior attitudes facilitate preparation for an upcoming persuasive message, general action goals were hypothesized to facilitate conscious retrieval of prior attitudes and therefore hinder attitude change to a greater extent than general inaction goals. Experiment 1 demonstrated that action primes (e.g., "go," "energy") yielded faster attitude report than inaction primes (e.g., "rest," "still") among participants who were forewarned of an upcoming persuasive message. Experiment 2 showed that the faster attitude report identified in Experiment 1 was localized on attitudes toward a message topic participants were prepared to receive. Experiments 3, 4, and 5 showed that, compared with inaction primes, action primes produced less attitude change and less argument scrutiny in response to a counterattitudinal message on a previously forewarned topic. Experiment 6 confirmed that the effects of the primes on attitude change were due to differential attitude retrieval. That is, when attitude expression was induced immediately after the primes, action and inaction goals produced similar amounts of attitude change. In contrast, when no attitude expression was induced after the prime, action goals produced less attitude change than inaction goals. Finally, Experiment 7 validated the assumption that these goal effects can be reduced or reversed when the goals have already been satisfied by an intervening task. PMID- 21639652 TI - Clients' perceptions of their psychotherapists' multicultural orientation. AB - The current retrospective study examined whether clients' (N = 176) perceptions of their psychotherapists' multicultural orientation (MCO) were associated with their psychological functioning, working alliance, and real relationship scores. Moreover, we tested whether clients' perceptions of the working alliance and the real relationship mediated the relationship between clients' perceptions of their psychotherapists' MCO and psychological functioning. The results showed that clients' perceptions of their psychotherapists' MCO were positively related to working alliance, real relationship, and psychological functioning. Only clients' ratings of the working alliance mediated the relationship between clients' perceptions of their psychotherapists' MCO and psychological functioning. Thus, because clients perceive their psychotherapists as being more oriented toward cultural issues, they may view the therapist as being more credible and may gain a sense of comfort in the therapeutic process. In turn, clients' strong alliance facilitates improvement in psychological well-being. PMID- 21639653 TI - Utilizing technological innovations to enhance psychotherapy supervision, training, and outcomes. AB - Recent technological advances in the use of the Internet and video technologies has greatly impacted the provision of psychotherapy and other clinical services as well as how the training of psychotherapists may be conducted. When utilized appropriately these technologies may provide greater access to needed services to include treatment, consultation, supervision, and training. Specific ethical challenges and pitfalls are discussed and recommendations are made for the ethical use of these technologies. Additionally, innovative practices from the seven articles in the special section that follows are highlighted and reviewed. These articles present a number of innovations that can take psychotherapy training, research, supervision, and treatment forward toward increased effectiveness. Recommendations for integrating these innovations into ongoing practices are provided and for additional research to build on the important work of the authors in this special section are provided. PMID- 21639654 TI - Web-conference supervision for advanced psychotherapy training: a practical guide. AB - The advent of readily accessible, inexpensive Web-conferencing applications has opened the door for distance psychotherapy supervision, using video recordings of treated clients. Although relatively new, this method of supervision is advantageous given the ease of use and low cost of various Internet applications. This method allows periodic supervision from point to point around the world, with no travel costs and no long gaps between direct training contacts. Web conferencing permits face-to-face training so that the learner and supervisor can read each other's emotional responses while reviewing case material. It allows group learning from direct supervision to complement local peer-to-peer learning methods. In this article, we describe the relevant literature on this type of learning method, the practical points in its utilization, its limitations, and its benefits. PMID- 21639655 TI - Learning psychotherapy in the digital age. AB - There is substantial literature documenting the process factors that lead to effective psychotherapy. Similarly, there is now a wealth of data attesting to the effectiveness of several psychotherapy brands. Little is known about the elements that facilitate learning how to be an effective clinician. One important step, after reading about a treatment model and seeing techniques demonstrated, is having the chance to practice the approach and receiving feedback and coaching from an experienced, knowledgeable supervisor. To accomplish this efficiently, most programs rely not only on trainee accounts of what went on in their therapy sessions, but also on recordings and videos of therapeutic encounters. This article describes our experience over a 5-year period in developing the use of Webcams for training psychology interns and psychiatric residents in the delivery of psychotherapy services. Pragmatic and technical details are given about how we went about establishing a recording system that is easy to use and provides secure, confidential storage of information at a reasonable cost. Discussion addresses both the weighing of choices that need to be made and overcoming the hesitation of trainees to reveal their work during treatment sessions. PMID- 21639656 TI - Learning how to rate video-recorded therapy sessions: a practical guide for trainees and advanced clinicians. AB - Watching and rating psychotherapy sessions is an important yet often overlooked component of psychotherapy training. This article provides a simple and straightforward guide for using one Website (www.ATOStrainer.com) that provides an automated training protocol for rating of psychotherapy sessions. By the end of the article, readers will be able to have the knowledge to go to the Website and begin using this training method as soon as they have a recorded session to view. This article presents, (a) an overview of the Achievement of Therapeutic Objectives Scale (ATOS; McCullough et al., 2003a), a research tool used to rate psychotherapy sessions; (b) a description of APA training tapes, available for purchase from APA Books, that have been rated and scored by ATOS trained clinicians and posted on the Website; (c) step-by-step procedures on how ratings can be done; (d) an introduction to www.ATOStrainer.com where ratings can be entered and compared with expert ratings; and (e) first-hand personal experiences of the authors using this training method and the benefits it affords both trainees and experienced therapists. This psychotherapy training Website has the potential to be a key resource tool for graduate students, researchers, and clinicians. Our long-range goal is to promote the growth of our understanding of psychotherapy and to improve the quality of psychotherapy provided for patients. PMID- 21639657 TI - Using the e-SOFTA for video training and research on alliance-related behavior. AB - In this article, we describe a specific technology for training/supervision and research on the working alliance in either individual or couple/family therapy. The technology is based on the System for Observing Family Therapy Alliances (SOFTA; Friedlander, Escudero, & Heatherington, 2006), which contains four conceptual dimensions (Engagement in the Therapeutic Process, Emotional Connection with the Therapist, Safety within the Therapeutic System, and Shared Sense of Purpose within the Family), observational rating tools (SOFTA-o), and self-report measures (SOFTA-s) shown to be important indicators of therapeutic progress. The technology, e-SOFTA, is a computer program (available for PC downloading free of charge) that can be used to rate client(s) and therapist on the specific SOFTA-o behaviors that contribute to or detract from a strong working alliance in each dimension. In addition to providing time-stamped frequencies of alliance-related behaviors, e-SOFTA allows users to link the observed behaviors to qualitative comments and to compare one person's rating of a session to that of another person. Suggestions are provided for using e-SOFTA in research, in didactic training, and in supervision, including a specific training module for introducing graduate students to the working alliance and assessing their observational and executive skills. PMID- 21639658 TI - Preliminary results of a video-assisted psychotherapist workshop in alliance strategies. AB - This study examined the preliminary results of an integrative, video-assisted training workshop aimed at helping psychotherapists build strong therapeutic relationships with their clients. Participants were 57 clinicians across five community mental health clinics, who were randomly assigned to the brief alliance training workshop (in which they participated prior to starting treatment with a new client) or to a delayed-training control condition. Outcomes assessed included therapist-reported use of alliance strategies during Session 1, therapist-rated alliance quality after Session 1, and client engagement across the first 4 weeks. In contrast to hypotheses, one-way analyses of variance and chi-square analyses revealed no statistically significant differences between the training and the delayed-training conditions. However, the therapist-reported impact of using the workshop's alliance strategies was positively correlated with therapist-rated alliance quality (r = .30, p = .03) and marginally correlated with number of sessions attended (r = .25, p = .06) across the two conditions. The findings hold promise for the utility of a brief alliance-focused workshop, and for collaborations between researchers and clinicians seeking to bridge science and practice. PMID- 21639659 TI - Effects of video-based therapy preparation targeting experiential acceptance or the therapeutic alliance. AB - Preparation for psychotherapy may enhance the psychotherapeutic process, reduce drop-outs, and improve outcomes, but the effective mechanisms of such preparation are poorly understood. Previous studies have rarely targeted specific processes that are associated with positive therapy outcomes. This randomized experiment compared the effects of preparatory videos that targeted either the Therapeutic Alliance, Experiential Acceptance, or a Control video on early therapeutic process variables in 105 patients seen in individual therapy. Participants watched the videos just before their first therapy session. No significant differences were found between the Alliance and Experiential Acceptance videos on patient recommendations, immediate affective reactions, or working alliance and attrition after the first session. However, the Therapeutic Alliance video produced an immediate increase in negative mood relative to the Control video, whereas the Experiential acceptance video produced a slight increase in positive mood relative to the Alliance video. Surprisingly, patients who viewed the Alliance video were rated significantly lower than the control group on therapist rated alliance after the first session. These findings suggest there may be specific process effects in the early phase of treatment based on the type of pretraining material used, and also indicate that video-based pretraining efforts could be counterproductive. Furthermore, this research contributes to the literature by providing insights into methodological considerations for future work on the use of technology in psychotherapy and challenges associated with preparing people for successful psychotherapy. PMID- 21639660 TI - Video-tape role induction for psychotherapy: moving forward. AB - Role induction (RI) has been shown to decrease premature termination and to enhance the therapeutic alliance and symptom relief. We examine the effects of a video-tape RI on premature termination rates, outcome, and the process variable of therapeutic alliance. Sixty-eight clients and their therapists (N = 26) participated in the study. Each client completed baseline measures of symptom status before beginning the therapy. Clients were then randomly assigned to either a video-tape RI condition or a no videotape treatment-as-usual condition. After the first session of therapy, clients and therapists separately completed outcome and alliance measures. Providing a video-tape RI to clients as they entered therapy did not lead to better results in attendance, outcome, or process variables. Considering possibilities that lie beyond the design of this study, so as to understand these results, is encouraged in an effort to advance the field's thinking about RI and how it can best be harnessed for continued positive effects moving forward. PMID- 21639661 TI - Internet and video technology in psychotherapy supervision and training. AB - The seven articles in this special section on the use of Internet and video technology represent the latest growth on one branch of the increasingly prolific and differentiated work in the technology of psychotherapy. In addition to the work presented here on video and the Internet applications to supervision and training, information technology is changing the field of psychotherapy through computer assisted therapies and virtual reality interventions. PMID- 21639662 TI - Emerging and continuing trends in psychotherapy: views from an editor's eye. AB - It is proposed that six major trends in psychotherapy have continued or emerged over the course of the author's editorship of Psychotherapy, the past seven years. These trends are (a) the increasing integration of techniques and the therapeutic relationship; (b) increasing focus on theoretical integration; (c) increasing efforts at research-practice integration; (d) increases in more specific, integrative reviews; (e) integration of biological, neuroscience understandings; and (f) integration of diversity and cultural considerations into psychotherapy. Each trend is described and its impact on the field is discussed. Cautions about each trend are also noted. The six trends are discussed in the context of integration. PMID- 21639663 TI - Trauma-focused cognitive behavioral therapy of a child with posttraumatic stress disorder. AB - This case study involves the use of Trauma-Focused Cognitive Behavioral Therapy to treat a preadolescent male patient referred to the Delaware public mental health system due to a history of family violence and symptoms associated with Posttraumatic Stress Disorder. Pre- to post-treatment data on self- and parent report measures demonstrate symptom reduction and exemplify the effectiveness of the model. Data on parent participation in the session and facilitation of trauma discussion at home illustrate the parent's contribution to the therapeutic process. Excerpts of clinical dialogue between child, parent, and therapist highlight the capacity of the model to accommodate individual needs and circumstances. Clinical recommendations supplement the treatment manual and provide clinicians with practical information for use in their own practices. PMID- 21639664 TI - What are the benefits of mindfulness? A practice review of psychotherapy-related research. AB - Research suggests that mindfulness practices offer psychotherapists a way to positively affect aspects of therapy that account for successful treatment. This paper provides psychotherapists with a synthesis of the empirically supported advantages of mindfulness. Definitions of mindfulness and evidence-based interpersonal, affective, and intrapersonal benefits of mindfulness are presented. Research on therapists who meditate and client outcomes of therapists who meditate are reviewed. Implications for practice, research, and training are discussed. PMID- 21639668 TI - Corvid caching: Insights from a cognitive model. AB - Caching and recovery of food by corvids is well-studied, but some ambiguous results remain. To help clarify these, we built a computational cognitive model. It is inspired by similar models built for humans, and it assumes that memory strength depends on frequency and recency of use. We compared our model's behavior to that of real birds in previously published experiments. Our model successfully replicated the outcomes of two experiments on recovery behavior and two experiments on cache site choice. Our "virtual birds" reproduced declines in recovery accuracy across sessions, revisits to previously emptied cache sites, a lack of correlation between caching and recovery order, and a preference for caching in safe locations. The model also produced two new explanations. First, that Clark's nutcrackers may become less accurate as recovery progresses not because of differential memory for different cache sites, as was once assumed, but because of chance effects. And second, that Western scrub jays may choose their cache sites not on the basis of negative recovery experiences only, as was previously thought, but on the basis of positive recovery experiences instead. Alternatively, both "punishment" and "reward" may be playing a role. We conclude with a set of new insights, a testable prediction, and directions for future work. PMID- 21639669 TI - Sequentially simulated outcomes: kind experience versus nontransparent description. AB - Recently, researchers have investigated differences in decision making based on description and experience. We address the issue of when experience-based judgments of probability are more accurate than are those based on description. If description is well understood ("transparent") and experience is misleading ("wicked"), it is preferable to experience. However, if description is not transparent, will valid ("kind") experience lead to more accurate judgments? We report 2 experiments. The first involved 7 well-known probabilistic inference tasks. Participants differed in statistical sophistication and answered with and without experience obtained through sequentially simulated outcomes. The second experiment involved interpreting the outcomes of a regression analysis when making inferences for investment decisions. In both experiments, even the statistically naive achieved accurate probabilistic inferences after experiencing sequentially simulated outcomes, and many preferred this presentation format. We conclude by discussing theoretical and practical implications. PMID- 21639670 TI - Culture, attention, and emotion. AB - This research provides experimental evidence for cultural influence on one of the most basic elements of emotional processing: attention to positive versus negative stimuli. To this end, we focused on Russian culture, which is characterized by brooding and melancholy. In Study 1, Russians spent significantly more time looking at negative than positive pictures, whereas Americans did not show this tendency. In Study 2, Russian Latvians were randomly primed with symbols of each culture, after which we measured the speed of recognition for positive versus negative trait words. Biculturals were significantly faster in recognizing negative words (as compared with baseline) when primed with Russian versus Latvian cultural symbols. Greater identification with Russian culture facilitated this effect. We provide a theoretical discussion of mental processes underlying cultural differences in emotion research. PMID- 21639671 TI - Parallel consolidation of simple features into visual short-term memory. AB - Although considerable research has examined the storage limits of visual short term memory (VSTM), little is known about the initial formation (i.e., the consolidation) of VSTM representations. A few previous studies have estimated the capacity of consolidation to be one item at a time. Here we used a sequential simultaneous manipulation to reexamine the limits of consolidating items into VSTM. Participants viewed briefly presented and masked color patches (targets), which were shown either sequentially or simultaneously. A probe color followed the targets and participants decided whether it matched one of the targets or was a novel color. In four experiments, we consistently found equal performance for sequential and simultaneous presentations for two targets. Worse performance in the simultaneous than the sequential condition was observed for larger set sizes (three and four). Contrary to previous results, suggesting a severe capacity limit of one item, our results indicate that consolidation into VSTM can occur in parallel and without capacity limits for at least two items. PMID- 21639673 TI - Perceptual and decisional factors influencing the discrimination of inversion in the Thatcher illusion. AB - The Thatcher illusion (Thompson, 1980) is considered to be a prototypical illustration of the notion that face perception is dependent on configural processes and representations. We explored this idea by examining the relative contributions of perceptual and decisional processes to the ability of observers to identify the orientation of two classes of forms-faces and churches-and a set of their component features. Observers were presented with upright and inverted images of faces and churches in which the components (eyes, mouth, windows, doors) were presented either upright or inverted. Observers first rated the subjective grotesqueness of all of the images and then performed a complete identification task in which they had to identify the orientation of the overall form and the orientation of each of the interior features. Grotesqueness ratings for both classes of image showed the standard modulation of rated grotesqueness as a function of orientation. The complete identification results revealed violations of both perceptual and decisional separability but failed to reveal any violations of within-stimulus (perceptual) independence. In addition, exploration of a simple bivariate Gaussian signal detection model of the relationship between identification performance and judged grotesqueness suggests that within-stimulus violations of perceptual independence on their own are insufficient for producing the illusion. This lack of evidence for within stimulus configurality suggests the need for a critical reevaluation of the role of configural processing in the Thatcher illusion. (PsycINFO Database Record (c) 2011 APA, all rights reserved). PMID- 21639674 TI - Chasing vs. stalking: interrupting the perception of animacy. AB - Visual experience involves not only physical features such as color and shape, but also higher-level properties such as animacy and goal-directedness. Perceiving animacy is an inherently dynamic experience, in part because agents' goal-directed behavior may be frequently in flux-unlike many of their physical properties. How does the visual system maintain and update representations of agents' animate and goal-directed behavior over time and motion? The present study explored this question in the context of a particularly salient form of perceived animacy: chasing, in which one shape (the "wolf") pursues another shape (the "sheep"). Here the participants themselves controlled the movement of the sheep, and the perception of chasing was assessed in terms of their ability to avoid being caught by the wolf-which looked identical to many moving distractors, and so could be identified only by its motion. The wolf's pursuit was frequently interrupted by periods in which it was static, jiggling in place, or moving randomly (amidst distractors that behaved similarly). Only the latter condition greatly impaired the detection of chasing-and only when the random motion was grouped into temporally extended chunks. These results reveal (1) how the detection of chasing is determined by the character and temporal grouping (rather than just the brute amount) of "pursuit" over time; and (2) how these temporal dynamics can lead the visual system to either construct or actively reject interpretations of chasing. PMID- 21639676 TI - Modern alternative approaches to the problem of drug-induced liver injury. PMID- 21639675 TI - Transposed letter priming with horizontal and vertical text in Japanese and English readers. AB - It is well established that in masked priming, a target word (e.g., JUDGE) is primed more effectively by a transposed letter (TL) prime (e.g., jugde) than by an orthographic control prime (e.g., junpe). This is inconsistent with the slot coding schemes used in many models of visual word recognition. Several alternative coding schemes have been proposed in which special bigram detectors for frequently occurring nonadjacent letter combinations are developed as a product of perceptual learning. In order to examine this perceptual learning hypothesis, we asked whether bigram detectors are defined in terms of visuospatial coordinates. Japanese-English bilinguals who were equally familiar with horizontal and vertical text in Japanese demonstrated strong TL priming in both orientations when reading Japanese words, but, when reading English words, the evidence for vertical TL priming was not as strong. However, native English speakers showed a clear TL priming effect with vertically presented English words despite minimal exposure to vertical text, which is not consistent with a perceptual learning account. It is proposed instead that the initial letter array is transformed into an abstract ordinal code (first to last) regardless of orientation and that the speed with which this transformation is carried out depends on the familiarity of the script. PMID- 21639677 TI - Alternatives for lung research: stuck between a rat and a hard place. AB - The respiratory system acts as a portal into the human body for airborne materials, which may gain access via the administration of medicines or inadvertently during inhalation of ambient air (e.g. air pollution). The burden of lung disease has been continuously increasing, to the point where it now represents a major cause of human morbidity and mortality worldwide. In the UK, more people die from respiratory disease than from coronary heart disease or non respiratory cancer. For this reason alone, gaining an understanding of mechanisms of human lung biology, especially in injury and repair events, is now a principal focus within the field of respiratory medicine. Animal models are routinely used to investigate such events in the lung, but they do not truly reproduce the responses that occur in humans. Scientists committed to the more robust Three Rs principles of animal experimentation (Reduction, Refinement and Replacement) have been developing viable alternatives, derived from human medical waste tissues from patient donors, to generate in vitro models that resemble the in vivo human lung environment. In the specific case of inhalation toxicology, human-oriented models are especially warranted, given the new REACH regulations for the handling of chemicals, the rising air pollution problems and the availability of pharmaceutically valuable drugs. Advances in tissue-engineering have made it feasible and cost-effective to construct human tissue equivalents of the respiratory epithelia. The conducting airways of the lower respiratory system are a critical zone to recapitulate for use in inhalation toxicology. Three dimensional (3-D) tissue designs which make use of primary cells, provide more in vivo-like responses, based on the targeted interactions of multiple cell types supported on artificial scaffolds. These scaffolds emulate the native extracellular matrix, in which cells differentiate into a functional pulmonary tissue. When 3-D cell cultures are employed for testing aerosolised chemicals, drugs and xenobiotics, responses are captured that mirror the events in the in situ human lung and provide human endpoint data. PMID- 21639678 TI - The use of a chemistry-based profiler for covalent DNA binding in the development of chemical categories for read-across for genotoxicity. AB - An important molecular initiating event for genotoxicity is the ability of a compound to bind covalently with DNA. However, not all compounds that can undergo covalent binding mechanisms will result in genotoxicity. One approach to solving this problem, when in silico prediction techniques are being used, is to develop tools that allow chemicals to be grouped into categories based on their ability to bind covalently to DNA. For this analysis to take place, compounds need to be placed within categories where the trend in toxicity can be explained by simple descriptors, such as hydrophobicity. However, this can occur only when the compounds within a category are structurally and mechanistically similar. Chemistry-based profilers have the ability to screen compounds and highlight those with similar structures to a target compound, and are thus likely to act via a similar mechanism of action. Here, examples are reported to highlight how structure-based profilers can be used to form categories and hence fill data gaps. The importance of developing a well-defined and robust category is discussed in terms of both mechanisms of action and structural similarity. PMID- 21639679 TI - Toward preclinical predictive drug testing for metabolism and hepatotoxicity by using in vitro models derived from human embryonic stem cells and human cell lines - a report on the Vitrocellomics EU-project. AB - Drug-induced liver injury is a common reason for drug attrition in late clinical phases, and even for post-launch withdrawals. As a consequence, there is a broad consensus in the pharmaceutical industry, and within regulatory authorities, that a significant improvement of the current in vitro test methodologies for accurate assessment and prediction of such adverse effects is needed. For this purpose, appropriate in vivo-like hepatic in vitro models are necessary, in addition to novel sources of human hepatocytes. In this report, we describe recent and ongoing research toward the use of human embryonic stem cell (hESC)-derived hepatic cells, in conjunction with new and improved test methods, for evaluating drug metabolism and hepatotoxicity. Recent progress on the directed differentiation of human embryonic stem cells to the functional hepatic phenotype is reported, as well as the development and adaptation of bioreactors and toxicity assay technologies for the testing of hepatic cells. The aim of achieving a testing platform for metabolism and hepatotoxicity assessment, based on hESC-derived hepatic cells, has advanced markedly in the last 2-3 years. However, great challenges still remain, before such new test systems could be routinely used by the industry. In particular, we give an overview of results from the Vitrocellomics project (EU Framework 6) and discuss these in relation to the current state-of-the-art and the remaining difficulties, with suggestions on how to proceed before such in vitro systems can be implemented in industrial discovery and development settings and in regulatory acceptance. PMID- 21639680 TI - Evaluation of anti-inflammatory and atrophogenic effects of glucocorticoids on reconstructed human skin. AB - Topical glucocorticoids (GCs) are extensively used in the treatment of inflammatory skin diseases. However, their long-term use is often accompanied by severe and eventually irreversible adverse effects, with atrophy being the most important limitation. Currently, most non-clinical studies involve animal testing, so the results are not always representative of the situation in humans. The aim of this project was to establish an in vitro test protocol for the evaluation of the anti-inflammatory and atrophic potential of topically applied GCs in reconstructed human skin. Initial studies with fibroblasts and keratinocytes confirmed the anti-inflammatory and atrophogenic effects of GCs, as evidenced by decreased cytokine production and collagen mRNA expression. In non pretreated reconstructed human skin (EpiDermFTTM), the topical application of GCs for seven days strongly reduced the secretion of interleukin (IL)-6. GC-induced skin atrophy, known to appear only after prolonged treatment, was not detected by the analysis of epidermal thickness and collagen mRNA expression. However, reproducible epidermal inflammation was established for the first time in reconstructed human skin. Topical treatment with tumour necrosis factor (TNF) increased IL-6 release and strongly reduced epidermal thickness accompanied by severe parakeratosis. GC treatment of reconstructed human skin reduced IL-6 levels and completely resolved parakeratosis, leading to the normalisation of epidermal thickness. These induced inflammatory conditions mimic more closely the clinical situations in which GCs are used, and therefore appear to be more suitable for future investigations for the establishment of a human-based in vitro test protocol for evaluating wanted and unwanted GC effects. PMID- 21639681 TI - Implementation of the three Rs in the human hazard assessment of Brazilian medicinal plants: an evaluation of the cytotoxic and genotoxic potentials of Dipteryx alata Vogel. AB - In Brazil, medicinal plants are widely used by the indigenous people, which leads to a constant requirement for toxicity tests to be performed on the plant extracts. Although the current Brazilian Directive 90/2004 on the preclinical toxicity testing of phytotherapeutics recommends only in vivo tests, some Brazilian researchers would like to change this situation by implementing the Three Rs in the toxicological testing of medicinal plants. The present study evaluated the cytotoxic and genotoxic potentials of bark extracts from Dipteryx alata Vogel, a medicinal plant of the Brazilian cerrado, by using CHO-K1 (Chinese hamster ovary) cells. An IC50 value was obtained, which corresponded to 0.16mg/ml of plant extract, and from this the equivalent LD50 was determined as 705mg/kg. In order to determine the genotoxic potential of the sample, the frequency of micronucleus formation was assessed. CHO-K1 cells were exposed, during targeted mitosis, to different concentrations of plant extract and cytochalasin B, in the presence and absence of an appropriate metabolic activation system (an S9 mix). The results obtained indicated that it might be possible to implement the Three Rs in assessing the potential human hazard of medicinal plants. The publication of such data can increase awareness of the Three Rs by showing how to optimise the management of animal use, if in vivo toxicological experiments are required. PMID- 21639682 TI - Evaluation of the acute antihyperglycemic effects of four selected indigenous plants from Jordan used in traditional medicine. AB - CONTEXT: Eryngium creticum Lam. (Umbelliferae), Geranium graveolens L.Her.exn Ait (Geraniaceae), Paronychia argentea Lam. (Caryophyllaceae), and Varthemia iphionoides Boiss (Compositae) have traditionally been used as antidiabetic phytomedicines. However, their alleged benefits and mechanisms remain elusive. OBJECTIVES: To evaluate the effect of these plants on in vitro and in vivo enzymatic starch digestion. MATERIALS AND METHODS: In vitro enzymatic starch digestion with acarbose or (1-50 or 100 mg/ml) plants aqueous extracts was assayed using alpha-amylase and alpha-amyloglucosidase. Oral starch tolerance tests and oral glucose tolerance tests were determined for the plant extracts at concentrations 125, 250, and 500 mg/kg body weight. Blood glucose levels in rats treated with plant extracts or drugs (acarbose or metformin and glipizide) were measured at -30, 0, 45, 90, and 135 min. RESULTS AND DISCUSSION: In vitro, acarbose, and water extracts of G. graveolens and V. iphionoides exerted significant dose-dependent dual inhibition of alpha-amylase and alpha glucosidase, with respective IC50s of 1.2 MUg/ml, 84.7, and 65.2 mg/ml. Comparable in vivo acute postprandial antihyperglycemic efficacies were obtained for G. graveolens and V. iphionoides in starch-fed rats. E. creticum exhibited substantial acute antihyperglycemic activities in starch-treated rats, despite lacking any favorable in vitro effectiveness. However, P. argentea lacked any inhibitory efficacy. None of the plant extracts qualified for improving the glucose tolerance in fasted rats on glucose loading. CONCLUSION: G. graveolens and V. iphionoides can be considered as potential candidates for therapeutic modulation of impaired fasting glycemia, impaired glucose tolerance, and type 2 diabetes. PMID- 21639683 TI - Cuminum cyminum extract attenuates scopolamine-induced memory loss and stress induced urinary biochemical changes in rats: a noninvasive biochemical approach. AB - CONTEXT: Cuminum cyminum Linn. (Apiaceae), cumin, is a popular spice with a long history of medicinal use to treat various symptoms such as diarrhea, flatulence, gynecological, and respiratory diseases. OBJECTIVE: To date, no scientific investigation was reported regarding memory-enhancing and antistress activity of cumin fruits. The present study deals with the memory-enhancing and antistress activities and further the antioxidant status via lipid peroxidation inhibition. MATERIALS AND METHODS: Antistress activity was evaluated by inducing stress via forced swimming and the urinary vanillylmandelic acid (VMA) and ascorbic acid were estimated as biomarkers. Memory-enhancing activity was studied by conditioned avoidance response using Cook's pole climbing apparatus in normal and scopolamine-induced amnestic rats. Thiobarbituric acid reactive substances (TBARS) assay was used to evaluate the lipid peroxidation. RESULTS: Daily administration of cumin at doses of 100, 200, and 300 mg/kg body weight 1 h prior to induction of stress inhibited the stress-induced urinary biochemical changes in a dose-dependent manner without altering the levels in normal control groups. The cognition, as determined by the acquisition, retention, and recovery in rats, was observed to be dose-dependent. The extract also produced significant lipid peroxidation inhibition in comparison with known antioxidant ascorbic acid in both rat liver and brain. DISCUSSION AND CONCLUSION: This study provides scientific support for the antistress, antioxidant, and memory-enhancing activities of cumin extract and substantiates that its traditional use as a culinary spice in foods is beneficial and scientific in combating stress and related disorders. PMID- 21639684 TI - Antioxidant activity of Citrus limon essential oil in mouse hippocampus. AB - CONTEXT: Citrus limon (L.) Burms (Rutaceae) has been shown in previous studies to have various biological functions (anti-inflammatory, antiallergic, antiviral, antimutagenic, and anticarcinogenic). However, traditional uses in folk medicine suggest that C. limon may have an effect on the central nervous system (CNS). OBJECTIVE: This study investigated the effects of C. limon essential oil (EO) on lipid peroxidation level, nitrite content, glutathione reduced (GSH) concentration, and antioxidant enzymes [superoxide dismutase (SOD), catalase, and glutathione peroxidase (GPx)] activities in mice hippocampus. MATERIALS AND METHODS: Swiss mice were treated with the suspension of 0.5% Tween 80, in distilled water used as vehicle (i.p., control group) and with EO in three different doses (0.05, 0.1, or 0.15 g/kg, i.p., EO 50, EO 100, and EO 150 groups, respectively). After the treatments, all groups were observed for 24 h. The enzyme activities as well as the lipid peroxidation, nitrite, and GSH concentrations in mice hippocampus were measured using spectrophotometric methods and the results were compared with values obtained from control group. RESULTS: EO of C. limon treatment significantly reduced the lipid peroxidation level and nitrite content but increased the GSH levels and the SOD, catalase, and GPx activities in mice hippocampus. DISCUSSION AND CONCLUSION: Our findings strongly support the hypothesis that oxidative stress in hippocampus can occur during neurodegenerative diseases, proving that hippocampal damage induced by the oxidative process plays a crucial role in brain disorders, and also imply that a strong protective effect could be achieved using EO of C. limon as an antioxidant. PMID- 21639685 TI - Inhibition of 5-lipoxygenase and cyclooxygenase-2 pathways by pain-relieving plaster in macrophages. AB - CONTEXT: Pain-relieving plaster (PRP) is a traditional Chinese medicine (TCM) that has been widely used with satisfactory results in the treatment of some diseases related to inflammation, such as bruises, chronic arthritis. OBJECTIVE: The mechanisms underlying the anti-inflammatory actions of PRP are investigated in this study for the first time. MATERIALS AND METHODS: The anti-inflammatory effects of PRP extracts were evaluated in lipopolysaccharide (LPS) or calcium ionophore A23187-treated murine peritoneal macrophages (PMs). Tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), prostaglandin E2 (PGE2), and leukotrienes B4 (LTB4) were evaluated by ELISA assays. Reverse transcriptase polymerase chain reaction (RT-PCR) and western blot analysis were used to detect the expression of cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX). Nuclear factor-kappa B (NF-kappaB)-DNA-binding activity was determined by gel mobility shift assay. RESULTS: PRP extracts were found to inhibit the production of TNF alpha, IL-1beta, and PGE(2), reduce the expressions of COX-2 at the mRNA and protein levels induced by LPS, and reduced the production of LTB4 induced by A23187. Furthermore, PRP extracts significantly attenuated LPS-induced NF-kappaB DNA-binding activity. DISCUSSION AND CONCLUSION: The anti-inflammatory effects of PRP possibly are related to reduction of inflammatory cytokines (TNF-alpha and IL 1beta), inducible inflammatory enzyme (COX-2), and its metabolite PGE2 via NF kappaB signal pathway. Moreover, PRP extracts also notably inhibited the production of LTB4, indicating that PRP inhibited the 5-LOX pathway, which may be the other mechanism for its anti-inflammatory action. PMID- 21639686 TI - Extraction of brown pigment from Rosa laevigata and its antioxidant activities. AB - CONTEXT: Rosa laevigata Michx. (Rosaceae), widespread in China, contains many valuable nutrients and has long been used as food and medicine in Chinese folklore. Nowadays, due to its favorable property of coloring, the brown pigment of R. laevigata has an attractive potential as an available additive in food. OBJECTIVE: The aims of this study were to optimize the extraction process of brown pigment from R. laevigata and investigate its antioxidant activities on the basis of its abilities to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, hydroxyl radical and superoxide radical. MATERIALS AND METHODS: Extraction conditions of brown pigment from R. laevigata were investigated through an orthogonal design of L9(3)4 assay. Ethanol concentration, extraction temperature, time, and ratio of material to solvent were the main factors affecting the extraction rate. Subsequently, the antioxidant activity of brown pigment was assessed using DPPH method, while hydroxyl radicals and superoxide free radicals were respectively determined by the Fenton-RhB (Rhodamine B) system and using the pyrogallol-luminol system. RESULTS: The optimum extraction conditions were determined: temperature 70 degrees C, ethanol concentration was 60%, extraction time 2 h and ratio of material to solvent was 1:6. Brown pigment showed a good radical scavenging activity, and exhibited a concentration-dependent inhibition of hydroxyl radical and superoxide free radical at low concentrations. When the concentration of brown pigment was 1 mg/mL, the scavenging percentage of hydroxyl radical reached 67.33%. DISCUSSION AND CONCLUSION: The brown pigment of R. laevigata could potentially be used as a promising natural antioxidant in the food and pharmaceutical industries. PMID- 21639687 TI - Morphogenetic changes in essential oil composition of Hypericum perforatum during the course of ontogenesis. AB - CONTEXT: In the past few years, an increasing interest in the volatile secondary metabolites of Hypericum perforatum L. (Guttiferae) has been arising. OBJECTIVE: The present study is a contribution to better understand the relationship between the morphological variations and volatile composition during the phenological cycle. MATERIALS AND METHODS: Leaves at the stages of vegetative, floral budding, flowering and green capsule, buds, full opened flowers and green capsules were assayed for essential oil (EO) components by gas chromatography-flame ionization detector (GC-FID) and GC-mass spectrometry (MS). RESULTS: Significant amounts of sesquiterpenes (oxygenated 26-50% and hydrocarbons 20-40%) and oxygenated hydrocarbons (13-38%) characterized the all analyzed samples showing peculiar fluctuations during the seven phenological stages. Although monoterpenes were present in much lower amounts (monoterpene hydrocarbons 0.4-6%; oxygenated monoterpenes 0.8-6%) they were considered also important discrimination for several stages. The green capsules and the full opened flowers collected at flowering stage were clearly distinguished in terms of EO compositions from the other samples. DISCUSSION: For the first time, the EO composition of Turkish wild Hypericum perforatum was monitored by the hydrodistillation of different plant organs collected at different seven stages in order to point out the modification of target volatiles related to each phenological step. CONCLUSIONS: Based on the EO composition monitored during these seven morphological stages by GC-MS, principal component analysis and cluster analysis, significant metabolite modifications were observed during the phenological cycle which involved the levels of specific volatile target compounds belonging to the chemical classes of hydrocarbons, monoterpenes and sesquiterpenes. PMID- 21639688 TI - Inhibition of urease by extracts derived from 15 Chinese medicinal herbs. AB - CONTEXT: Helicobacter pylori is a major causative factor in gastritis-like disorders, and urease plays a key role in Helicobacter pylori colonizing and persisting in the mucous layer of the human stomach. In China, a variety of Chinese medicinal herbs have been prescribed to attenuate or eradicate gastritis like disorders. However, little is known about the urease inhibition of Chinese medicinal herbs. OBJECTIVE: The present study was conducted to investigate the urease inhibition activities of the ethanol and water extracts of 15 Chinese medicinal herbs. MATERIALS AND METHODS: The ethanol and water extracts derived from 15 medicinal herbs, traditionally used for the treatment of gastritis-like disorders in China, were tested for urease-inhibition activity using the phenol red method. RESULTS: Screened at 10 ug/mL, 14 ethanol extracts and 10 water extracts showed urease inhibition. The ethanol extracts of Magnolia officinalis Rehd. et Wils. (Magnoliaceae) and Cassia obtusifolia L. (Leguminosae) possessed inhibition rates higher than 50% with IC50 values of 6.5 and 12.3 ug/mL, respectively. After fractionating successively, the petroleum ether fraction of the ethanol extracts of Magnolia officinalis showed the best activity with 90.8% urease inhibition at a concentration of 10 ug/mL. The bioautography of the petroleum ether fraction indicated the existence of the urease inhibitors in the herb. DISCUSSION AND CONCLUSION: The present results indicated that some Chinese medicinal herbs might treat gastritis-like disorders via the inhibition of Helicobacter pylori urease and the further possibility for discovering useful novel urease inhibitors from the Chinese medicinal herbs. PMID- 21639689 TI - Chemical constituents and bioactivities of the liposoluble fraction from different medicinal parts of Crocus sativus. AB - CONTEXT: Crocus sativus Linn. (Iridaceae), commonly known as saffron, becomes more and more popular due to its versatile biological and medicinal properties. At present, studies mainly focus on the traditional medicinal part, the saffron stigma, with less attention to the other parts of saffron, such as the perianth, the stamen, and the corm, which are high yield compared to the stigma and also possess various pharmacological effects. OBJECTIVE: To determine the chemical compositions, antifungal, cytotoxic, and antioxidant activities of the ether fractions from the stamen, perianth, and stigma of saffron. MATERIALS AND METHODS: The chemical constituents of the ether fractions from different parts of saffron were investigated by gas chromatography/mass spectrometry. Several pathogenic fungi isolates and tumor cell lines were employed to evaluate the antifungal and cytotoxic activities of these three ether fractions. 1,1-Diphenyl 2-picrylhydrazyl assay was used to determine the free radical-scavenging activity. RESULTS: The ether fractions composition of the three C. sativus parts are different from each other, but lauric acid, hexadecanoic acid, 4 hydroxydihydro-2(3H)-furanone, and stigmasterol were the common constituents shared by all the three fractions. The stamen ether fraction displayed the strongest antifungal and cytotoxic activities, whereas both of the saffron stamen and perianth ether fractions exhibited significant antioxidant activities. DISCUSSION AND CONCLUSION: These findings demonstrate that the saffron stamen and perianth possess significant antifungal, cytotoxic, and antioxidant activities as well as the stigma, though not to the same extent, prompting us to expand the medicinal resource and make best use of this valuable plant. PMID- 21639690 TI - Inhibitory effects of essential oil from rhizomes of Ligusticum chuanxiong on hypertrophic scarring in the rabbit ear model. AB - CONTEXT: Hypertrophic scarring, a common proliferative disorder of dermal fibroblasts, results from an overproduction of collagen and excessive deposition of extracellular matrix. Although the treatment with surgical excisions or steroid hormones can modify the symptoms, numerous treatment-related complications have also been established. OBJECTIVE: To investigate the effects of essential oil (EO) from rhizomes of Ligusticum chuanxiong Hort. (Umbelliferae) on hypertrophic scarring in a rabbit ear model. MATERIALS AND METHODS: A rabbit ear model of hypertrophic scarring was established. EO (5, 10, and 20%) was applied once daily to the scars for 22 days. After 28 days of post-wounding, excision of scars was respectively performed for both histological examination and assays of the levels of collagen I, collagen III, matrix metalloproteinase-1 (MMP-1), and transforming growth factor beta 1 (TGF-beta1). The scar elevation index (SEI) was also determined. RESULTS: After 22 days of treatment with indicated concentrations of EO, hypertrophic scarring was significantly inhibited in the rabbit ears. The levels of TGF-beta1, collagen I, and collagen III evidently decreased and MMP-1 level markedly increased in the scar tissue. SEI was also significantly reduced. Immunohistochemical findings exhibited significant amelioration of the scar tissue. DISCUSSION AND CONCLUSION: EO suppresses hypertrophic scarring in the rabbit ear model and is a probably effective cure for human hypertrophic scarring. PMID- 21639691 TI - Influence of Carbopol 71G-NF on the release of dextromethorphan hydrobromide from extended-release matrix tablets. AB - The objective of this study was to evaluate the potential of Carbopol((r)) 71G-NF on the release of dextromethorphan hydrobromide (DM) from matrix tablets in comparison with hydroxypropyl methylcellulose (HPMC((r)) K15M) and Eudragit((r)) L100-55 polymers. Controlled release DM matrix tablets were prepared using Carbopol 71G-NF, HPMC K15M, and Eudragit L100-55 at different drug to polymer ratios by direct compression technique. The mechanical properties of the tablets as tested by crushing strength and friability tests were improved as the concentration of Carbopol, HPMC, and Eudragit increased. However, Carbopol-based tablets showed a significantly (P<0.05) higher crushing strength and a lower friability than HPMC and Eudragit tablets. No significant differences in weight uniformity and thickness values were observed between the different formulations. It was also found that Carbopol significantly (P<0.05) delayed the release of DM in comparison with HPMC K15M and Eudragit L100-55. A combination of HPMC K15M and Eudragit L100-55 in a 1:1 ratio at 20 and 30% significantly (P<0.05) delayed the release of DM than Eudragit L100-55 alone. Moreover, blends of Carbopol and HPMC at a 1:1 ratio at the 10, 20, and 30% total polymer concentration were investigated. The blend of Carbopol and HPMC at 10% level significantly (P<0.05) slowed the release of DM than Carbopol or HPMC alone, whereas blends at 20 and 30% level significantly (P<0.05) delayed the release of DM compared with HPMC or Carbopol alone. The results with these polymer blends showed that it was possible to reduce the total amount of polymers when used as a combination in formulation. PMID- 21639692 TI - Toxicological evaluation of realistic emission source aerosols (TERESA): introduction and overview. AB - Determining the health impacts of sources and components of fine particulate matter (PM(2.5)) is an important scientific goal. PM(2.5) is a complex mixture of inorganic and organic constituents that are likely to differ in their potential to cause adverse health outcomes. The Toxicological Evaluation of Realistic Emissions of Source Aerosols (TERESA) study focused on two PM sources--coal-fired power plants and mobile sources--and sought to investigate the toxicological effects of exposure to emissions from these sources. The set of papers published here document the power plant experiments. TERESA attempted to delineate health effects of primary particles, secondary (aged) particles, and mixtures of these with common atmospheric constituents. TERESA involved withdrawal of emissions from the stacks of three coal-fired power plants in the United States. The emissions were aged and atmospherically transformed in a mobile laboratory simulating downwind power plant plume processing. Toxicological evaluations were carried out in laboratory rats exposed to different emission scenarios with extensive exposure characterization. The approach employed in TERESA was ambitious and innovative. Technical challenges included the development of stack sampling technology that prevented condensation of water vapor from the power plant exhaust during sampling and transfer, while minimizing losses of primary particles; development and optimization of a photochemical chamber to provide an aged aerosol for animal exposures; development and evaluation of a denuder system to remove excess gaseous components; and development of a mobile toxicology laboratory. This paper provides an overview of the conceptual framework, design, and methods employed in the study. PMID- 21639693 TI - Toxicological evaluation of realistic emission source aerosols (TERESA)--power plant studies: assessment of breathing pattern. AB - Our approach to study multi-pollutant aerosols isolates a single emissions source, evaluates the toxicity of primary and secondary particles derived from this source, and simulates chemical reactions that occur in the atmosphere after emission. Three U.S. coal-fired power plants utilizing different coals and with different emission controls were evaluated. Secondary organic aerosol (SOA) derived from alpha-pinene and/or ammonia was added in some experiments. Male Sprague-Dawley rats were exposed for 6 h to filtered air or different atmospheric mixtures. Scenarios studied at each plant included the following: primary particles (P); secondary (oxidized) particles (PO); oxidized particles + SOA (POS); and oxidized and neutralized particles + SOA (PONS); additional control scenarios were also studied. Continuous respiratory data were obtained during exposures using whole body plethysmography chambers. Of the 12 respiratory outcomes assessed, each had statistically significant changes at some plant and with some of the 4 scenarios. The most robust outcomes were found with exposure to the PO scenario (increased respiratory frequency with decreases in inspiratory and expiratory time); and the PONS scenario (decreased peak expiratory flow and expiratory flow at 50%). PONS findings were most strongly associated with ammonium, neutralized sulfate, and elemental carbon (EC) in univariate analyses, but only with EC in multivariate analyses. Control scenario O (oxidized without primary particles) had similar changes to PO. Adjusted R(2) analyses showed that scenario was a better predictor of respiratory responses than individual components, suggesting that the complex atmospheric mixture was responsible for respiratory effects. PMID- 21639694 TI - Syringomyelia. PMID- 21639695 TI - Supracerebellar approach. PMID- 21639696 TI - Detection of brain micrometastases by high-resolution stereotactic magnetic resonance imaging and its impact on the timing of and risk for distant recurrences. AB - OBJECT: The aim of this study was to assess the order of micrometastases that can be detected with high-resolution MR imaging at the time of Gamma Knife surgery (GKS), and to estimate the impact this has on the time until and incidence of distant recurrences. METHODS: A consecutive series of 835 patients with brain metastases treated with GKS in a 7-year period, excluding patients in whom earlier brain metastases were treated with other modalities, were retrospectively analyzed. In all patients GKS was based on high Gd-dose (0.3 mmol/kg), high resolution stereotactic MR imaging. These images were compared with the standard pretreatment MR images, and the difference in number of metastases found was analyzed. The distant recurrence rate following GKS was compared with that found in a prospective randomized study (Aoyama et al.) comparing radiosurgery to radiosurgery plus prophylactic whole-brain radiation therapy. RESULTS: New tumors were diagnosed in 40% (95% CI 36%-43%) of all patients as well as in the majority of patients with multiple lesions found on the diagnostic scan. The more tumors there were on the diagnostic scan, the higher the likelihood of detecting additional lesions with high-resolution imaging. It was calculated that approximately 50% of the micrometastases present at the time of GKS could be diagnosed with high-resolution imaging, which decreased the incidence of and delayed the time for the development of distant recurrences. CONCLUSIONS: Additional brain metastases can be diagnosed in 40% of patients by using high resolution imaging. Thus, radiosurgical treatments based on high-resolution stereotactic MR imaging decrease the incidence of and lengthen the time to distant recurrences. PMID- 21639697 TI - Familial nontraumatic, nonaneurysmal subarachnoid hemorrhage: a report on three first-degree siblings. AB - Family history is a recognized risk factor in aneurysmal subarachnoid hemorrhage (SAH). The genetic and environmental contributions are actively researched. The authors of this report present a case series of 3 first-degree siblings affected by nontraumatic, angiographically negative SAH. Data in this study suggest that familial predisposition may also apply to spontaneous, nonaneurysmal SAH and that family history should be actively investigated in all such patients. The identification of families with multiple affected members could lead to an improved understanding of the genetic and environmental factors associated with this condition. PMID- 21639698 TI - Prophylactic antiepileptic drug therapy in patients undergoing supratentorial meningioma resection: a systematic analysis of efficacy. AB - OBJECT: Meningiomas are one of the more common intracranial neoplasms. The risk of seizures and secondary aspiration, brain edema, and brain injury often leads practitioners to administer prophylactic antiepileptic drugs (AEDs) perioperatively. The efficacy of this practice remains controversial, however, with prior investigations reaching conflicting results and recent studies focusing on AED side effects. The authors performed a systematic analysis of outcomes following supratentorial meningioma resection with and without prophylactic AED administration in the hope of clarifying the role of AEDs in the perioperative care of patients with these lesions. METHODS: A MEDLINE search of the literature (1979-2010) was performed. Comparisons were made for patient and tumor characteristics as well as success of repair, morbidity, and seizure outcome. Statistical analyses of categorical variables were undertaken using chi square and Fisher exact tests. RESULTS: Nineteen studies, involving 698 patients, were included. There were no significant differences in the extent of resection, perioperative mortality, or recurrence between the AED and no-AED cohorts. Likewise, there were no significant differences in the incidence of early or late seizures between the cohorts. CONCLUSIONS: The results of this systematic analysis supports the conclusion that the prophylactic administration of anticonvulsants during resection of supratentorial meningiomas provides no benefit in the prevention of either early or late postoperative seizures. Despite their traditional role in this patient population, the routine use of AEDs should be carefully reconsidered. PMID- 21639699 TI - Postoperative antiepileptic drugs. PMID- 21639700 TI - Intraoperative magnetic resonance imaging and early prognosis for vision after transsphenoidal surgery for sellar lesions. AB - OBJECT: Sellar lesions with suprasellar extension may cause loss of visual acuity and visual field damage due to compression of the optic chiasm. Using intraoperative MR (iMR) imaging to detect symptomatic lesion remnants adjacent to the optic chiasm (that may be resected in the same procedure) may positively affect the functional outcome of patients with these lesions. The aim of this study was to evaluate the correlation between visual improvement and optic nerve decompression detected by iMR imaging in patients undergoing transsphenoidal resection of pituitary lesions. METHODS: A total of 32 patients (23 men and 9 women) who underwent transsphenoidal resection of sellar lesions causing visual impairment were included in this study. Tumor volume ranged from 0.9 cm(3) to 55.7 cm(3) (mean 9.8 +/- 11.7 cm(3)). Preoperative assessment showed visual field damage in 31 patients (97%) and loss of visual acuity in 28 patients (88%). The latency period between the appearance of symptoms and transsphenoidal decompression was 14.9 +/- 19.5 weeks. RESULTS: Intraoperative MR imaging was performed after the resection was believed to be complete, or if further tumor removal was not safely possible due to changed conditions in the surgical field. Complete resection was detected on these initial scans in 17 patients (53%). Partial resection was achieved in 9 patients (28%) and tumor debulking in 6 (19%). Additional resection was possible in 8 (53%) of these 15 patients. Four (50%) of these 8 cases had suprasellar remnants and the optic chiasm was subsequently decompressed. In 5 cases optimal decompression of the optic chiasm was not possible. On early follow-up within 1 month after surgery, overall improvement of visual field damage was observed in 27 patients (87%). In 23 patients (74%), the Goldmann perimetry demonstrated complete recovery. Improvement of visual acuity was noted in 24 patients (86%). Eighteen patients (64%) regained full visual acuity. Identification of a decompressed optic chiasm on iMR imaging was significantly correlated with visual field improvement (p = 0.0007; positive predictive value 0.96, 95% CI 0.81-0.99) and relief of visual acuity deficits (p = 0.0002; positive predictive value 0.96, 95% CI 0.79-0.99). Two patients needed transcranial procedures for symptomatic tumor remnants detected on iMR imaging. CONCLUSIONS: Intraoperative MR imaging findings correlate with prognosis of visual deficits after transsphenoidal decompression of the anterior optic pathways. The use of iMR imaging may prevent revision surgery for unexpected symptomatic remnants. PMID- 21639701 TI - Computed tomography-based morphometric analysis of the human occipital condyle for occipital condyle-cervical fusion. AB - OBJECT: Occipital condyle screws serve as an alternative fixation point in occipital-cervical fusion. Their placement requires a thorough understanding of the anatomy of the occipital condyles and associated structures. This study is a CT-based morphometric analysis of occipital condyles as related to occipital condyle-cervical fusion. METHODS: A total of 170 patients were examined with CT scans of the craniocervical junction at a single institution, for a total of 340 occipital condyles, between March 6, 2006, and July 30, 2006. All CT scans were negative for traumatic, degenerative, and neoplastic pathological entities. Condylar anteroposterior (AP) length, transverse width, height, projected screw angle, and projected screw lengths were measured on an EBW Portal 2.5 CT Viewer Workstation (Philips Electronics). The longest axis in the AP orientation of the occipital condyle was accepted as the length. The transverse width was a line perpendicular to the midpoint of the long axis. The height was measured in the coronal projection that had the thickest craniocaudal portion of the condyle. The screw trajectory started 5 mm lateral to the medial edge of the condyle and a line was directed anteromedially in the longest axis. The angle was measured relative to the sagittal midline. The screw length was measured from the outer cortex of the posterior wall to the outer cortex of the anterior wall. RESULTS: The mean +/- SD values for occipital condyle measurements were as follows: AP length was 22.38 +/- 2.19 mm (range 14.7-27.6 mm); width was 11.18 +/- 1.44 mm (range 7.4-19.0 mm); height was 9.92 +/- 1.30 mm (range 5.1-14.3 mm); screw angle was 20.30 degrees +/- 4.89 degrees (range 8.0 degrees -34.0 degrees ); and screw length was 20.30 +/- 2.24 mm (range 13.0-27.6 mm). CONCLUSIONS: These measurements correlate with previous cadaveric and radiographic studies of the occipital condyle, and emphasize the role of preoperative planning for the feasibility of placement of an occipital condyle screw. PMID- 21639702 TI - Late instrumentation failure after total en bloc spondylectomy. AB - OBJECT: The object of this study was to investigate failures after spinal reconstruction following total en bloc spondylectomy (TES), related factors, and sequelae arising from such failures in patients with malignant spinal tumors. METHODS: Fifteen patients (12 males and 3 females, with a mean age of 46.5 years) with malignant spinal tumors who underwent TES and survived for more than 1 year were included in this analysis (mean follow-up 41.5 months). Seven patients had primary tumors, including giant cell tumors in 4 patients, chordoma in 2, and Ewing sarcoma in 1. Eight patients had metastatic tumors, including thyroid cancer in 6 and renal cell cancer and malignant fibrous histiocytoma in 1 patient each. Seven patients without prominent paravertebral extension of the tumor were treated using a posterior approach alone, and 8 patients who exhibited prominent anterior or anterolateral extension of the tumors into the thoracic or abdominal cavity were treated using a combined anterior and posterior approach. Spinal reconstruction after tumor resection was performed using a combination of anterior structural support and posterior instrumentation. The relationship between instrumentation failure and clinical and radiographic factors, including age, sex, history of previous surgery, preoperative radiotherapy, tumor histology, tumor level, surgical approach, number of resected vertebrae, rod diameter, number of instrumented vertebrae, and cage subsidence, was investigated. RESULTS: Six patients (40%) with spinal instrumentation failure were identified: rod breakage occurred in 3 patients, and breakage of both the rod and the cage, combined cage breakage and screw back-out, and endplate fracture arising from cage subsidence occurred in 1 patient each. All of these patients experienced acute or chronic back pain, but only 1 patient with a tumor recurrence experienced neurological deterioration upon instrumentation failure. Cage subsidence (>= 5 mm), preoperative irradiation, and the number of instrumented vertebrae (<= 4 vertebrae) were significantly related to late instrumentation failure. CONCLUSIONS: Late instrumentation failure was a frequent complication after TES. Although patients with instrumentation failure experienced back pain, the neurological sequelae were not catastrophic. For prevention, meticulous preparation of the graft site and a longer posterior fixation should be considered. PMID- 21639703 TI - Severe onychodystrophy caused by allergic contact dermatitis to acrylates in artificial nails. AB - Acrylic resin monomers, especially acrylates and methacrylates, are well-known sensitizers responsible for allergic contact dermatitis mainly in the occupational setting. The most frequently affected professionals are dentists, orthopedic surgeons, manicurists, painting industry and fiberglass workers. The authors report the case of a 39-year-old healthy woman, a secretary, who developed severe onychodystrophy of all fingers, 1 week after the application of sculptured acrylic nails. PMID- 21639704 TI - More on periorbital infections following therapy with biological agents. PMID- 21639705 TI - Haplotype combination of SREBP-1c gene sequence variants is associated with growth traits in cattle. AB - The aim of this study was to examine the association of the SREBP-1c polymorphism with growth traits in cattle breeds. Five sequence variants (SVs) were identified within the bovine sterol regulatory element-binding protein-1c gene (SREBP-1c), using DNA sequencing, PCR, PCR-RFLP, and forced PCR-RFLP methods. These polymorphisms include three missense mutations (SV1, SV4, and SV5) in exons 7, 9, and 12, a silent mutation (SV3) in exon 9, and a large deletion (SV2) in intron 7. Overall, we report the validation of polymorphisms within the bovine SREBP-1c gene, and the haplotype variability and extent of linkage disequilibrium (LD) in 1061 individuals representing the five main cattle breeds from China. We also investigated haplotype frequencies and LD coefficients for five SVs in all study populations. LD and haplotype structure of SREBP-1c were different between breeds. The result of haplotype analysis of five SVs showed that 27 different haplotypes were identified by all breeds. Two haplotypes (Hap1 and Hap2) shared by all five populations accounted for 42.75%, 35.68%, 36.44%, 25.43%, and 96.26% of all haplotypes observed in the cattle breeds Nanyang, Qinchuan, Jiaxian, Jinnan, and Chinese Holstein, respectively. The statistical analyses indicated that one single SV and 38 combined haplotypes were significantly associated with growth traits in the Nanyang cattle population (P < 0.05 or P < 0.01). The results of this study suggest that the SREBP-1c gene possibly is a strong candidate gene that affects growth traits in the Chinese beef cattle breeding program. PMID- 21639706 TI - Molecular and cellular mechanism of lung injuries due to exposure to sulfur mustard: a review. AB - Sulfur mustard (SM), a potent chemical weapon agent, was used by Iraqi forces against Iranian in the Iraq-Iran war (1981-1989). Chronic obstructive pulmonary disease (COPD) is a late toxic pulmonary consequence after SM exposure. The COPD observed in these patients is unique (described as Mustard Lung) and to some extent different from COPD resulted from other well-known causes. Several mechanisms have been hypothesized to contribute to the pathogenesis of COPD including oxidative stress, disruption of the balance between apoptosis and replenishment, proteinase-antiproteinase imbalance and inflammation. However, it is not obvious which of these pathways are relevant to the pathogenesis of mustard lung. In this paper, we reviewed studies addressing the pathogenicity of mustard lung, and reduced some recent ambiguities in this field. There is ample evidence in favor of crucial role of both oxidative stress and apoptosis as two known mechanisms that are more involved in pathogenesis of mustard lung comparing to COPD. However, according to available evidences there are no such considerable data supporting neither proteolytic activity nor inflammation mechanism as the main underlying pathogenesis in Mustard Lung. PMID- 21639707 TI - Quantification of the pathological response and fate in the lung and pleura of chrysotile in combination with fine particles compared to amosite-asbestos following short-term inhalation exposure. AB - The marked difference in biopersistence and pathological response between chrysotile and amphibole asbestos has been well documented. This study is unique in that it has examined a commercial chrysotile product that was used as a joint compound. The pathological response was quantified in the lung and translocation of fibers to and pathological response in the pleural cavity determined. This paper presents the final results from the study. Rats were exposed by inhalation 6 h/day for 5 days to a well-defined fiber aerosol. Subgroups were examined through 1 year. The translocation to and pathological response in the pleura was examined by scanning electron microscopy and confocal microscopy (CM) using noninvasive methods. The number and size of fibers was quantified using transmission electron microscopy and CM. This is the first study to use such techniques to characterize fiber translocation to and the response of the pleural cavity. Amosite fibers were found to remain partly or fully imbedded in the interstitial space through 1 year and quickly produced granulomas (0 days) and interstitial fibrosis (28 days). Amosite fibers were observed penetrating the visceral pleural wall and were found on the parietal pleural within 7 days postexposure with a concomitant inflammatory response seen by 14 days. Pleural fibrin deposition, fibrosis, and adhesions were observed, similar to that reported in humans in response to amphibole asbestos. No cellular or inflammatory response was observed in the lung or the pleural cavity in response to the chrysotile and sanded particles (CSP) exposure. These results provide confirmation of the important differences between CSP and amphibole asbestos. PMID- 21639708 TI - Flow cytometry of sputum: assessing inflammation and immune response elements in the bronchial airways. AB - BACKGROUND: The evaluation of sputum leukocytes by flow cytometry (FCM) is an opportunity to assess characteristics of cells residing in the central airways, yet it is hampered by certain inherent properties of sputum including mucus and large amounts of contaminating cells and debris. OBJECTIVE: To develop a gating strategy based on specific antibody panels in combination with light scatter properties for flow cytometric evaluation of sputum cells. METHODS: Healthy and mild asthmatic volunteers underwent sputum induction. Manually selected mucus "plug" material was treated with dithiothreitol, filtered and total leukocytes acquired. Multicolor FCM was performed using specific gating strategies based on light scatter properties, differential expression of CD45 and cell lineage markers to discriminate leukocytes from squamous epithelial cells and debris. RESULTS: The combination of forward scatter and CD45 expression reliably segregated sputum leukocytes from contaminating squamous epithelial cells and debris. Overlap of major leukocyte populations (neutrophils, macrophages/monocytes) required the use of specific antibodies (e.g. CD16, CD64, CD14, HLA-DR) that differentiated granulocytes from monocytes and macrophages. These gating strategies allowed identification of small populations of eosinophils, CD11c+ myeloid dendritic cells, B-cells and natural killer cells. CONCLUSIONS: Multicolor FCM can be successfully applied to sputum samples to identify and characterize leukocyte populations residing on the surfaces of the central airways. CLINICAL RELEVANCE: This research describes detailed methods to overcome difficulties associated with FCM of sputum samples, which previously has been lacking in the literature. FCM of sputum samples can provide valuable information on inflammation and immunological response elements in the bronchial airways for both clinical diagnostic and research applications and can be a useful tool in inhalation toxicology for assessing health effects of inhaled environmental pollutants. PMID- 21639709 TI - Pathological features of rat lung following inhalation and intratracheal instillation of C(60) fullerene. AB - We evaluated the pulmonary pathological features of rats that received a single intratracheal instillation and a 4-week inhalation of a fullerene. We used fullerene C(60) (nanom purple; Frontier Carbon Co. Ltd, Japan) in this study. Male Wistar rats received intratracheal dose of 0.1, 0.2, or 1 mg of C(60), and were sacrificed at 3 days, 1 week, 1 month, 3 months, 6 months, and 12 months. In the inhalation study, Wistar rats received C(60) or nickel oxide by whole-body inhalation for 6 h/day, 5 days/week, 4 weeks, and were sacrificed at 3 days, 1 month, and 3 months after the end of exposure. During the observation period, no tumors or granulomas were observed in either study. Histopathological evaluation by the point counting method (PCM) showed that a high dose of C(60) (1 mg) instillation led to a significant increase of areas of inflammation in the early phase (until 1 week). In the inhalation study of the C(60)-exposed group, PCM evaluation showed significant changes in the C(60)-exposed group only at 3 days after exposure; after 1 month, no significant changes were observed. The present study demonstrated that the pulmonary inflammation pattern after exposure to well characterized C(60) via both intratracheal and inhalation instillation was slight and transient. These results support our previous studies that showed C(60) has no significant adverse effects in intratracheal and inhalation instillation studies. PMID- 21639710 TI - PM2.5-induced changes in cardiac function of hypertensive rats depend on wind direction and specific sources in Steubenville, Ohio. AB - BACKGROUND: Increases in particulate matter less than 2.5 um (PM(2.5)) in ambient air is linked to acute cardiovascular morbidity and mortality. Specific components and potential emission sources of PM(2.5) responsible for adverse health effects of cardiovascular function are unclear. METHODS: Spontaneously hypertensive rats were implemented with radiotelemeters to record ECG responses during inhalation exposure to concentrated ambient particles (CAPs) for 13 consecutive days in Steubenville, OH. Changes in heart rate (HR) and its variability (HRV) were compared to PM(2.5) trace elements in 30-min time frames to capture acute physiological responses with real-time fluctuations in PM(2.5) composition. Using positive matrix factorization, six major source factors were identified: (i) coal/secondary, (ii) mobile sources, (iii) metal coating/processing, (iv) iron/steel manufacturing, (v) lead and (vi) incineration. RESULTS: Exposure-related changes in HR and HRV were dependant on winds predominately from either the northeast (NE) or southwest (SW). During SW winds, the metal processing factor was associated with increased HR, whereas factors of incineration, lead and iron/steel with NE winds were associated with decreased HR. Decreased SDNN was dominated during NE winds by the incinerator factor, and with SW winds by the metal factor. Metals and mobile source factors also had minor impacts on decreased SDNN with NE winds. Individual elemental components loaded onto these factors generally showed significant associations, although there were some discrepancies. CONCLUSIONS: Acute cardiovascular changes in response to ambient PM(2.5) exposure can be attributed to specific PM constituents and sources linked with incineration, metal processing, and iron/steel production. PMID- 21639711 TI - Particle-associated organic compounds and symptoms in myocardial infarction survivors. AB - CONTEXT: The aerosol components responsible for the adverse health effects of the exposure to particulate matter (PM) have not been conclusively identified, and there is especially little information on the role of particulate organic compounds (POC). OBJECTIVE: This study evaluated the role of PM and POC with regard to daily symptoms. METHODS: One hundred and fifty-three myocardial infarction survivors from Augsburg, Germany, recorded daily occurrence of different symptoms in winter 2003/2004. Ambient concentrations of PM with a diameter <2.5 MUm (PM(2.5)), particle number concentration (PNC), PM(2.5)-bound hopanes, and polycyclic aromatic hydrocarbons (PAH) were quantified. Data were analyzed using generalized estimating equations adjusting for meteorological and other time-variant confounders. RESULTS: The odds for avoidance of physically demanding activities due to heart problems increased immediately associated with most POC measures (e.g. 5% per 1.08 ng/m(3) increase in benzo[a]pyrene, 95% confidence interval (CI):1-9%) and tended to a delayed decrease. After a 2-day delayed decrease associated with hopanes, the odds for shortness of breath increased consistently after 3 days with almost all POC measures (e.g. 4% per 0.21 ng/m(3) increase in 17alpha(H), 21beta(H)-hopane, CI: 1-8%). The odds for heart palpitations marginally increased immediately in association with PNC (8% per 8146 cm(-3) increase in PNC, CI: 0-16%). CONCLUSIONS: The study showed an association between PM, particle-bound POC, and daily symptoms. The organic compounds may be causally related with cardiovascular health or act rather as indicators for traffic- and combustion-related particles. PMID- 21639712 TI - The PERFECT project: measuring performance of health care episodes. PMID- 21639713 TI - Analysing current trends in care of acute myocardial infarction using PERFECT data. AB - INTRODUCTION: This article in the supplement issue on the Performance, Effectiveness, and Costs of Treatment episodes (PERFECT)-project describes the PERFECT AMI (acute myocardial infarction) Database, which is developed to measure the performance of hospitals and hospital districts in Finland. We analyse annual trends and regional differences in performance indicators and whether the utilisation of services and costs of hospital care are related to improvement in survival of AMI patients. MATERIAL AND METHODS: The study population consists of ten annual cohorts (1998-2007) of patients hospitalised for AMI. RESULTS: Since 1998 the treatment pattern has changed rather radically, the utilisation rate of percutaneous coronary intervention (PCI) has increased and coronary procedures have been performed earlier after myocardial infarction. Outcome measured by various measures of mortality has improved considerably. However, trends in the development of the use of services and outcomes are not similar between hospital districts. An increase in cost was positively and statistically significantly related to decrease in mortality, but the effect was not very strong. CONCLUSION: There is potential for decreased mortality from actions that do not increase the costs and for enhancing performance in the regions and hospitals with poor performance. PMID- 21639714 TI - Trends in treatment and outcome of stroke patients in Finland from 1999 to 2007. PERFECT Stroke, a nationwide register study. AB - INTRODUCTION: This article in this supplement issue on the Performance, Effectiveness, and Costs of Treatment episodes (PERFECT) project describes trends in Finnish stroke treatment and outcome. MATERIAL AND METHODS: The PERFECT Stroke study uses multiple national registry linkages at individual patient level to produce a national stroke database with comprehensive follow-up of all hospital treated stroke patients in Finland. RESULTS: There were 94,316 incident stroke patients treated in Finnish hospitals from 1999 to 2007. Lengths-of-stays decreased after ischemic stroke (IS), and increased after intracerebral (ICH) and subarachnoid (SAH) hemorrhage. Ten-year survival improved in IS (hazard ratio 0.75; 95% CI 0.71-0.79) and ICH patients (0.88; 0.79-0.97), increasing median survival by 2 and 1 life-years respectively. This has translated into more days spent home among IS patients, but not among ICH patients. Treatment by neurologists improved the survival of IS (odds ratio [OR] 1.77; 95% CI 1.70-1.84) and ICH patients (OR 1.55; 95% CI 1.40-1.69), and treatment by neurosurgeons of SAH patients (OR 2.66; 95% CI 2.25-3.16), the effects were further improved by care in specialized stroke centers. DISCUSSION: The survival of Finnish IS and ICH patients has improved. Specialized acute care was associated with improved outcome. PMID- 21639715 TI - Regional and hospital variance in performance of total hip and knee replacements: a national population-based study. AB - INTRODUCTION: This article in the supplement on the Performance, Effectiveness, and Costs of Treatment episodes (PERFECT)-project describes the PERFECT Hip and Knee Replacement Database and its possibilities by evaluating regional and hospital-level differences in length of stay (LOS), costs and complication rates of total hip arthroplasty (THA) and total knee arthroplasty (TKA) in Finland. MATERIAL AND METHODS: All hip and knee arthroplasties are recorded in the Finnish Hospital Discharge Register (FHDR) and Finnish Arthroplasty Register (FAR). LOS, length of uninterrupted institutional care (LUIC), complication rates and other parameters of treatment were determined by region and hospital during 1998-2008 based on these. RESULTS: LOS and LUIC following THA and TKA diminished during the follow-up period. In 1998 average LOS after THA and TKA was 9.9 and 10 days. In 2008, these had shortened to 5.2 and 5.3 days, respectively. There was a 5.0 and 7.5 percentage point difference in revision rate between regions in THAs and TKAs, respectively, performed during 2005-2007 and followed to the end of 2009. DISCUSSION: The Finnish health care registers provide a monitoring system for evaluating hospital- and regional-level differences in THA and TKA. The differences in LOS, LUIC and revision rates between hospitals and regions are considerable. PMID- 21639716 TI - Monitoring the performance of hip fracture treatment in Finland. AB - INTRODUCTION: This article in the supplement on the PERFormance, Effectiveness, and Costs of Treatment episodes (PERFECT)-project aims to measure the performance and quality of hip fracture treatment by analysing annual trends and regional differences in developed performance indicators. MATERIAL AND METHODS: The PERFECT Hip Fracture Database contains all hip fracture patients identified from the Hospital Discharge Register in Finland since 1999. Follow-up data from several administrative registers were also linked to the database. Several risk adjusted performance indicators were developed. RESULTS: In 2007 (compared with 1999), 4.1 percentage points fewer patients had died and 7.5 percentage points more patients were at home four months after fracture. The mean length of treatment had shortened from about 50 to about 45 days, and the mean costs of treatment per patient during the year following hip fracture had increased from about ?18,000 to almost ?20,000. There was extensive variation between the hospitals in the proportion of patients with an operative delay longer than two days and clear differences between hospital districts in several performance indicators. CONCLUSIONS: Outcomes of hip fracture treatment in Finland have been improved in recent years, but regional variation exists. Register-based data are useful for performance assessment of hip fracture treatment. PMID- 21639717 TI - A methodological approach for register-based evaluation of cost and outcomes in health care. AB - INTRODUCTION: In health care, measures of performance are needed at producer level for improving the treatment processes and at system level for steering purposes. In addition, measures that enable reliable comparisons of producers with respect to each other should encourage them to develop their treatment processes to attain better positioning in benchmarking. METHODS: The main innovation of the Performance, Effectiveness, and Costs of Treatment episodes (PERFECT) project is to measure performance using existing linkable information available from registers within well-defined care episodes in a whole population. Finnish health care and related registers are used for constructing the disease specific databases, with rich content on treatment processes and complete follow up data. RESULTS: The PERFECT project has developed numerous performance indicators that can be used to evaluate health policy actions as well as to create regional and hospital-level benchmarking data. In PERFECT, the idea is to eliminate individual-level variation from the performance indicators by using individual-level data and proper risk adjustment methods. The focus of our interest is in the variation at the producer or regional level. CONCLUSIONS: Our experience shows that the utilization of population-level health care registers with an episode-of-care approach enables a continual system and producer-level performance measurement. PMID- 21639718 TI - PERFECT preterm infant study. AB - INTRODUCTION: This article summarizes the main findings of the preterm infant sub study of the Performance, Effectiveness and Costs of Treatment episodes (PERFECT) study. We studied effects of birth hospital level and time of birth on mortality and morbidity and cost-effectiveness of care of very low gestational age (VLGA)/very low birth weight (VLBW) infants. MATERIAL AND METHODS: The study included all infants born below 32 weeks or 1501 g in Finland in 2000-2007. Different cohorts were used depending on the time point. RESULTS: The one-year mortality of live-born VLBW/VLGA infants was higher if born in level II versus level III hospitals, or if born during out-of-office hours in level II versus office hours in level III hospitals. Two out of three VLGA/VLBW subjects did not have any of the prematurity-related morbidities studied. The average cost of quality-adjusted life years was ?19,245 by four years of age; the cost was higher in VLGA/VLBW infants with long-term morbidities. DISCUSSION: Birth in a level III hospital improved survival of VLGA/VLBW infants. Results suggest inadequate overnight competence in small hospitals. Despite high initial costs, care of VLGA/VLBW infants was already cost-effective by four years of age. Cost effectiveness can be improved by reducing long-term morbidities. PMID- 21639719 TI - PERFECT--conclusions and future developments. PMID- 21639720 TI - Effect of blast exposure on the brain structure and cognition in Macaca fascicularis. AB - Blast injury to the brain is one of the major causes of death and can also significantly affect cognition and physical and psychological skills in survivors of blast. The complex mechanisms via which blast injury causes impairment of cognition and other symptoms are poorly understood. In this study, we investigated the effects of varying degrees of primary blast overpressure (BOP; 80 and 200 kPa) on the pathophysiological and magnetic resonance imaging (MRI) changes and neurocognitive performance as assessed by the monkey Cambridge Neuropsychological Test Automated Battery (mCANTAB) in non-human primates (NHP). The study aimed to examine the effects of neurobehavioral and histopathological changes in NHP. MRI and histopathology revealed ultrastructural changes in the brain, notably in the Purkinje neurons in the cerebellum and pyramidal neurons in the hippocampus, which were most vulnerable to the blast. The results correlated well with the behavioral changes and changes in motor coordination and working memory of the affected monkeys. In addition, there was white matter damage affecting myelinated axons, astrocytic hypertrophy, and increased aquaporin-4 (AQP-4) expression in astrocytes, suggesting cerebral edema. Increased apoptosis appeared to involve astrocytes and oligodendrocytes in the animals following blast exposure. The small sample size could have contributed to the non significant outcome in cognitive performance post-blast and limited quantitative analyses. Nevertheless, the study has provided initial descriptive changes for establishing a primary BOP threshold for brain injury to serve as a useful platform for future investigations that aim to estimate brain injury potential and set safe limits of exposure. PMID- 21639721 TI - Rhythm disorders in isolated left ventricular noncompaction. AB - Both supraventricular and ventricular rhythm disorders are frequently observed in patients with isolated left ventricular noncompaction (IVNC). Most importantly, these patients are prone to develop life-threatening ventricular arrhythmias, which are amongst their most frequent causes of death. Data regarding risk stratification of ventricular arrhythmias, however, are scarce due to the rareness of the disease. Indeed, even invasive electrophysiological studies may be of limited value in this regard in the majority of patients. Implantable cardioverter defibrillators (ICDs) have been demonstrated to be highly effective for the prevention of sudden arrhythmic death in IVNC and should be considered in patients who are clinically judged to be at high risk for ventricular tachyarrhythmias. These include patients with a severely reduced ejection fraction as well as those with a prior history of sustained ventricular tachycardia or fibrillation, recurrent syncope of unknown etiology, or a family history of ventricular tachyarrhythmias or sudden cardiac death. This review summarizes the electrocardiographic and electrophysiological findings in patients with IVNC and discusses possibilities for risk stratification as well as the rationale for ICD implantation for the prevention of sudden cardiac death. PMID- 21639722 TI - Effect of aerobic training on hot flushes and quality of life--a randomized controlled trial. AB - BACKGROUND AND OBJECTIVE: To estimate whether aerobic training has an effect on frequency of hot flushes or quality of life. DESIGN: A randomized controlled trial. PARTICIPANTS AND SETTING: Symptomatic, sedentary women (n = 176), 43-63 years, no current use of hormone therapy. INTERVENTION: Unsupervised aerobic training for 50 minutes four times per week during 6 months. OUTCOMES: Hot flushes as measured with Women's Health Questionnaire (WHQ) and Health-Related Quality of Life (HRQoL, SF-36), daily reported hot flushes on phone-based diary, cardiorespiratory fitness (CRF), and body composition. RESULTS: Intervention group had larger decrease in the frequency of night-time hot flushes based on phone diary (P for month * group = 0.012), but not on WHQ scale. Intervention group had less depressed mood (P = 0.01) than control women according to change in WHQ score. Changes in WHQ score in depressed mood (P = 0.03) and menstrual symptoms (P = 0.01) in the intervention group were significantly dependent on frequency of training sessions. HRQoL was improved among the intervention group women in physical functioning (P = 0.049) and physical role limitation (P = 0.017). CRF improved (P = 0.008), and lean muscle mass increased (P = 0.046) significantly in the intervention group as compared to controls. CONCLUSIONS: Aerobic training may decrease the frequency of hot flushes and improve quality of life among slightly overweight women. PMID- 21639723 TI - Urinary incontinence: pharmacotherapy options. AB - The impact of incontinence is felt by millions of people worldwide, with tremendous decrement in quality of life and enormous cost reaching billions of dollars. Urinary incontinence is defined as 'involuntary leakage of urine' and is categorized into two main types: urgency urinary incontinence (UUI) and stress urinary incontinence (SUI). Behavioral modifications and pharmacologic therapies, primarily antimuscarinic agents, are the mainstay of treatment for UUI. These drugs are moderately efficacious but have troublesome side-effects, the combination resulting in poor compliance and persistence with therapy. There are several agents on the market today, each with some variation in pharmacologic properties. Whether these translate into meaningful differences in clinical efficacy and tolerability remains a matter of debate. Treatment of SUI has seen little success with pharmacologic therapy. In Europe, duloxetine is approved for treatment of SUI with marginal success rates; this drug, although available in the United States for treatment of depression, is not approved for SUI. The search for newer and better pharmacologic options and novel therapies is on going, fueled primarily by the high prevalence of bothersome incontinence and the tremendous number of health care dollars spent on current therapy. This review addresses pharmacologic options for treatment of urinary incontinence. PMID- 21639724 TI - Rodent model of direct cranial blast injury. AB - Traumatic brain injury resulting from an explosive blast is one of the most serious wounds suffered by warfighters, yet the effects of explosive blast overpressure directly impacting the head are poorly understood. We developed a rodent model of direct cranial blast injury (dcBI), in which a blast overpressure could be delivered exclusively to the head, precluding indirect brain injury via thoracic transmission of the blast wave. We constructed and validated a Cranium Only Blast Injury Apparatus (COBIA) to deliver blast overpressures generated by detonating .22 caliber cartridges of smokeless powder. Blast waveforms generated by COBIA replicated those recorded within armored vehicles penetrated by munitions. Lethal dcBI (LD(50) ~ 515 kPa) was associated with: (1) apparent brainstem failure, characterized by immediate opisthotonus and apnea leading to cardiac arrest that could not be overcome by cardiopulmonary resuscitation; (2) widespread subarachnoid hemorrhages without cortical contusions or intracerebral or intraventricular hemorrhages; and (3) no pulmonary abnormalities. Sub-lethal dcBI was associated with: (1) apnea lasting up to 15 sec, with transient abnormalities in oxygen saturation; (2) very few delayed deaths; (3) subarachnoid hemorrhages, especially in the path of the blast wave; (4) abnormal immunolabeling for IgG, cleaved caspase-3, and beta-amyloid precursor protein (beta-APP), and staining for Fluoro-Jade C, all in deep brain regions away from the subarachnoid hemorrhages, but in the path of the blast wave; and (5) abnormalities on the accelerating Rotarod that persisted for the 1 week period of observation. We conclude that exposure of the head alone to severe explosive blast predisposes to significant neurological dysfunction. PMID- 21639725 TI - Inhibition of enzyme activity by nanomaterials: potential mechanisms and implications for nanotoxicity testing. AB - The objective of this study was to investigate whether nanoparticle-exposure affects enzyme function and to determine the mechanisms responsible. Silicon, Au, and CdSe nanoparticles were synthesized in house and their physicochemical properties were characterized. The activity of purified lactate dehydrogenase (LDH) was inhibited or abolished by all nanoparticles tested. Inhibition was dependent upon particle core and surface-functional group composition. Inhibition of LDH was absent in crude tissue homogenates, in the presence of albumin, and at the isoelectric point of the protein, indicating that nanoparticles bind non specifically to abundant proteins via a charge interaction. Circular dichroism spectroscopy suggests that the structure of LDH may be altered by nanoparticles in a manner different from that of bulk controls. We present new data on the specific physicochemical properties of nanoparticles that may lead to bioactivity and highlight a number of potentially serious problems with common nanotoxicity testing methods. PMID- 21639726 TI - Organic solvents vapor pressure and relative humidity effects on the phase transition rate of alpha and beta forms of tegafur. AB - The objective of this work was to investigate the relative humidity (RH) and solvent vapor pressure effects on the phase transition dynamics between tegafur polymorphic forms that do not form hydrates and solvates. The commercially available alpha and beta modifications of 5-fluoro-1-(tetrahydro-2-furyl)-uracil, known as the antitumor agent tegafur, were used as model materials for this study. While investigating the phase transitions of alpha and beta tegafur under various partial pressures of methanol, n-propanol, n-butanol, and water vapor, it was determined that the phase transition rate increased in the presence of solvent vapors, even though no solvates were formed. By increasing the relative air humidity from 20% to 80%, the phase transition rate constant of alpha and beta tegafur was increased about 60 times. After increasing the partial pressure of methanol, n-propanol, or n-butanol vapor, the phase transition rate constant did not change, but the extent of phase transformation was increased. In the homologous row of n-alcohols, the phase transition rate constant decreased with increasing carbon chain length. The dependence of phase transformation extent versus the RH corresponded to the polymolecular adsorption isotherm with a possible capillary condensation effect. PMID- 21639727 TI - Detrimental effect of genetic inhibition of B-site APP-cleaving enzyme 1 on functional outcome after controlled cortical impact in young adult mice. AB - beta-Amyloid (Abeta) peptides, most notably associated with Alzheimer's disease, have been implicated in the pathogenesis of secondary injury after traumatic brain injury (TBI). A prior study has demonstrated that blocking the beta-site amyloid precursor protein (APP)-cleaving enzyme 1 (Bace1) required for production of Abeta from APP improved functional and histologic outcomes after controlled cortical impact (CCI) in aged mice. However, the majority of patients with severe TBI are young adults under the age of 40. Prior experimental models have suggested age-dependent differences in Abeta clearance, and a recent study in our lab suggests that young animals remediate acute elevations in Abeta after CCI better than older animals. We therefore tested the hypothesis that Bace1 deletion in young adult mice would not be protective after CCI. Male Bace1 knockout (Bace1(-/-)) and wild-type Bace1(+/+) (C57BL/6) mice (2-3 months old) were subjected to CCI (n=18-23/group) or sham injury (n=10-12/group). Functional outcomes were assessed with wire grip (motor) and the Morris water maze (MWM; spatial memory). Soluble Abeta levels were assessed at 48 h after CCI. Histopathological outcomes were assessed by lesion and hippocampal volume. Clustered ordinal logistic regression showed overall significant impairment in motor performance in injured Bace1(-/-) versus Bace1(+/+) animals (p=0.003). No significant differences in MWM performance were found on repeated-measures ANOVA (p=0.11) between groups. Probe scores were significantly worse in injured Bace1( /-) versus Bace1(+/+) mice (p=0.0009). Soluble Abeta(40) was significantly lower in ipsilateral hemispheres of Bace1(-/-) than in Bace1(+/+) animals after CCI (0.9 [IQR 0.88-0.94] pmol/g protein versus 3.8 [IQR 2.4-6.0] pmol/g protein; p=0.005). Lesion and hippocampal volumes did not differ between injured groups. The data suggest that therapies targeting Bace1 may need to be tailored according to age and injury severity, as their use may exacerbate functional deficits after TBI in younger or less severely injured patients. PMID- 21639728 TI - Decompressive craniectomy for elderly patients with traumatic brain injury: it's probably not worth the while. AB - Decompressive craniectomy (DC) has been regarded as an ultima ratio measure in the treatment of refractory intracranial hypertension after brain injury. Most discussion about its benefits is based on studies performed in patients who are <65 years of age. The aim of this study was to identify patients aged >=66 years who underwent DC after traumatic brain injury (TBI), in order to assess patient outcome and to correlate the values of potential predictors of survival on prognosis. From January 2002 to December 2009, 44 patients aged >=66 underwent DC (follow-up, 12-102 months). Potential predictors of outcome were analyzed, including age, post-resuscitation Glasgow Coma Scale (GCS) score, presence of mass lesion, Simplified Acute Physiology Score (SAPS) II, Injury Severity Score (ISS), and timing of surgical decompression. Mortality was 48% at discharge from the intensive care unit (ICU), 57% at hospital discharge, and 77% at 1-year follow-up and at last follow-up. A bad outcome Glasgow Outcome Scale Dead Vegetative State-Severely Disabled (GOS D-VS-SD) was observed in 36/44 patients both at hospital discharge and at 1-year follow-up. Mean SAPS II was 45.2 for patients who survived and 57.3 for patients who had died (p=0.0022). Patients who survived had a higher mean post-resuscitation GCS score (p=0.02). Logistical regression analysis indicated post-resuscitation GCS score as the only independent predictive factor for outcome. None of the 22 patients with a post resuscitation GCS score of 3-5 had a good outcome, 2/10 (20%) patients with a post-resuscitation GCS score of 6-8 and 6/12 patients (50%) with a post resuscitation GCS score >=9 had a good outcome. PMID- 21639729 TI - Interstitial F(2)-isoprostane 8-iso-PGF(2alpha) as a biomarker of oxidative stress after severe human traumatic brain injury. AB - Oxidative stress is a major contributor to the secondary injury process after experimental traumatic brain injury (TBI). The importance of oxidative stress in the pathobiology of human TBI is largely unknown. The F(2)-isoprostane 8-iso prostaglandin F(2alpha) (8-iso-PGF(2alpha)), synthesized in vivo through non enzymatic free radical catalyzed peroxidation of arachidonic acid, is a widely used biomarker of oxidative stress in multiple disease states, including TBI and cerebral ischemia/reperfusion. Our hypothesis is that harvesting of biomarkers directly in the injured brain by cerebral microdialysis (MD) is advantageous because of its high spatial and temporal resolution compared to blood or cerebrospinal fluid sampling. The aim of this study was to test the feasibility of measuring 8-iso-PGF(2alpha) in MD, ventricular cerebrospinal fluid (vCSF), and plasma samples collected from patients with severe TBI, and to compare the MD signals with MD-glycerol, implicated as a biomarker of oxidative stress, as well as MD-glutamate, a biomarker of excitotoxicity. Six patients (4 men, 2 women) were included in the study, three of whom had a focal/mixed TBI, and three a diffuse axonal injury (DAI). Following the bedside analysis of routine MD biomarkers (glucose, lactate:pyruvate ratio, glycerol, and glutamate), two 12-h MD samples per day were used to analyze 8-iso-PGF(2alpha) from 24 h up to 8 days post-injury. The interstitial levels of 8-iso-PGF(2alpha) were markedly higher than the levels obtained from plasma and vCSF (p<0.05), supporting our hypothesis. The MD-8-iso-PGF(2alpha) levels correlated strongly (p<0.05) with MD glycerol and MD-glutamate, which are widely used biomarkers of membrane phospholipid degradation/oxidative stress and excitotoxicity, respectively. This study demonstrates the feasibility of analyzing 8-iso-PGF(2alpha) in MD samples from the human brain. Our results support a close relationship between oxidative stress and excitotoxicity following human TBI. MD-8-iso-PGF(2alpha) in combination with MD-glycerol may be useful biomarkers of oxidative stress in the neurointensive care setting. PMID- 21639740 TI - Standard of practice and Flynn Effect testimony in death penalty cases. AB - The Flynn Effect is a well-established psychometric fact documenting substantial increases in measured intelligence test performance over time. Flynn's (1984) review of the literature established that Americans gain approximately 0.3 points per year or 3 points per decade in measured intelligence. The accurate assessment and interpretation of intellectual functioning becomes critical in death penalty cases that seek to determine whether an individual meets the criteria for intellectual disability and thereby is ineligible for execution under Atkins v. Virginia (2002) . We reviewed the literature on the Flynn Effect and demonstrated how failure to adjust intelligence test scores based on this phenomenon invalidates test scores and may be in violation of the Standards for Educational and Psychological Testing as well as the "Ethical Principles for Psychologists and Code of Conduct." Application of the Flynn Effect and score adjustments for obsolete norms clearly is supported by science and should be implemented by practicing psychologists. PMID- 21639741 TI - Diabetes among adults with cognitive limitations compared to individuals with no cognitive disabilities. AB - Using a retrospective analysis of data from the 2006 Medical Expenditures Panel Survey (MEPS), we assessed the health status of working-age adults with cognitive limitations in comparison to adults with no disability (unweighted N = 27,116; weighted N = 240,343,457). Adults with cognitive limitations had a significantly higher prevalence of diabetes than did adults with no disability (19.4% vs. 3.8%, respectively) and a significantly higher prevalence of six other major chronic conditions. In addition, individuals with cognitive limitations and diabetes were significantly more likely to have multiple (four or more) chronic illnesses. The health disparities we found in this study demonstrate the need to improve disease prevention and education efforts for individuals with cognitive limitations and their health care providers. PMID- 21639742 TI - Health status and health risks of the "hidden majority" of adults with intellectual disability. AB - Little is known about the health status of and health risks faced by adults with intellectual disability who do not use intellectual disability services. Self report data collected from 1,022 people with mild intellectual disability in England indicated that people who do not use intellectual disability services are more likely to smoke tobacco and less likely to access some health services and promotion activities than those who do use these services. In addition, they are more likely to be exposed to some known social determinants of poorer health (greater material hardship, greater neighborhood deprivation, reduced community, and social participation). PMID- 21639743 TI - Active support: a systematic review and evidence-based practice evaluation. AB - Researchers have evaluated active support in agencies for persons with developmental disabilities to increase staff assistance and service user engagement. A systematic review identified two studies in which researchers reported three experimental evaluations of active support. Only one experiment showed a clear functional relationship between active support with "ineffective" to "questionable" percentage of nonoverlapping data points effect sizes and acceptable percentage of all nonoverlapping data points effect sizes. Two experiments did not show experimental control; however, there was evidence that the investigators in these studies did not sufficiently manipulate the independent variable. Based on these data, active support only meets Chambless and Hollon's (1998) criterion for a "promising treatment" but not an evidence based practice. Future research on active support should demonstrate that the experimenter manipulated the independent variable and reported data on individual participants. PMID- 21639744 TI - Sowing the seeds of the autism field: Leo Kanner (1943). AB - More than 65 years after Leo Kanner published his seminal article, research on autism continues to be an area of increasing interest. Although much progress has been made, this field is still in its infancy, and many avenues of research are just beginning to be pursued. Despite the time that has passed, the syndrome Kanner identified and his comments about the children he observed continue to have meaning today, and although some of his suggestions about the etiology and presentation of autism were grounded in the thinking of his day, many of his observations were quite prescient. In this paper we explore Kanner's contributions to the field of autism, discuss how the field has changed, and suggest ways that research on autism spectrum disorders can continue to move forward. PMID- 21639745 TI - Colorado governor pardons Joe Arridy. PMID- 21639746 TI - Execution of Joe Arridy: comments of a forensic expert. PMID- 21639747 TI - Client evaluations and summaries: how person-centered planning is tainted by a diagnosis. PMID- 21639750 TI - Children and youth with intellectual or developmental disabilities living in congregate care settings (1977 to 2009): healthy people 2010 objective 6.7b outcomes (revised). PMID- 21639754 TI - A New Look For BI&T. PMID- 21639753 TI - Thalamus and cognitive impairment in mild traumatic brain injury: a diffusional kurtosis imaging study. AB - Conventional imaging is unable to detect damage that accounts for permanent cognitive impairment in patients with mild traumatic brain injury (mTBI). While diffusion tensor imaging (DTI) can help to detect diffuse axonal injury (DAI), it is a limited indicator of tissue complexity. It has also been suggested that the thalamus may play an important role in the development of clinical sequelae in mTBI. The purpose of this study was to determine if diffusional kurtosis imaging (DKI), a novel quantitative magnetic resonance imaging (MRI) technique, can provide early detection of damage in the thalamus and white matter (WM) of mTBI patients, and can help ascertain if thalamic injury is associated with cognitive impairment. Twenty-two mTBI patients and 14 controls underwent MRI and neuropsychological testing. Mean kurtosis (MK), fractional anisotropy (FA), and mean diffusivity (MD) were measured in the thalamus and several WM regions classically identified with DAI. Compared to controls, patients examined within 1 year after injury exhibited variously altered DTI- and DKI-derived measures in the thalamus and the internal capsule, while in addition to these regions, patients examined more than 1 year after injury also showed similar differences in the splenium of the corpus callosum and the centrum semiovale. Cognitive impairment was correlated with MK in the thalamus and the internal capsule. These findings suggest that combined use of DTI and DKI provides a more sensitive tool for identifying brain injury. In addition, MK in the thalamus might be useful for early prediction of permanent brain damage and cognitive outcome. PMID- 21639755 TI - The Roundup A compilation of items about medical device news, regulations, and AAMI initiatives. PMID- 21639756 TI - California medical instrumentation association. PMID- 21639758 TI - Probing the challenges of endoscopes. PMID- 21639757 TI - Ask george. PMID- 21639759 TI - Medical telemetry grows with changing nature of healthcare. PMID- 21639760 TI - Making your mark in the biomed world. PMID- 21639761 TI - Preamble to quality system regulation serves as valuable compliance tool. PMID- 21639762 TI - Creating a Cradle-to-Grave Asset Management Program. PMID- 21639763 TI - Slowly, healthcare getting greener. PMID- 21639764 TI - Radiation sterilization failures what can go wrong? PMID- 21639765 TI - Ann scott blouin. PMID- 21639766 TI - Right-sizing and replacing the right equipment at the right time. PMID- 21639767 TI - Why information management standards matter to biomeds. PMID- 21639768 TI - TENS Units. PMID- 21639769 TI - Solving problems with anesthesia machines and nerve stimulators. PMID- 21639770 TI - Cautions and precautions. PMID- 21639771 TI - A look at smart infusion pumps. PMID- 21639772 TI - Unified communications in healthcare. PMID- 21639773 TI - Did renovation project need renovation? PMID- 21639774 TI - How to save your department from cuts to training budget. PMID- 21639775 TI - Management and risk assessment of wireless medical devices in the hospital. PMID- 21639776 TI - Medical device interoperability a standards-based testing approach. AB - Abstract We present a black-box messaging test approach employed to achieve a level of rigor which improves-if not assures, given no optionality-correct data exchange. In particular, we verify that physiological information derived and communicated via messaging from a source medical device (e.g., an infusion pump) or healthcare information system, to another medical device (e.g., a patient monitor) or healthcare information system, which consumes or makes use of the data, is syntactically and semantically correct. In other words, the structure of information exchanged within the healthcare system is compliant to a defined specification(s) and the information meaning conveyed and interpreted by the consumer is exactly the same and as intended by the source. Our approach for developing a test system to validate messages is based on constraining identified and recognized specifications. The test system validation performed uses codified assertions derived from the specifications and constraints placed upon those specifications. To first show conformance-which subsequently enables interoperability-these assertions, which are atomic requirements traceable by clause to the base specifications, are employed by our medical device test tools to rigorously enforce standards to facilitate safe and effective plug-and-play information exchange. PMID- 21639777 TI - Are we too narrowly focused? PMID- 21639778 TI - Nonthrombogenic approaches to cardiovascular bioengineering. AB - Cardiovascular devices such as vascular grafts, stents, and heart valves have been widely used to treat cardiovascular diseases. The failure of these devices is usually initiated by the formation of thrombus and neointima on the device surfaces. Antithrombogenic surface modifications have been employed to improve the performance of these devices. In addition to biochemical modifications, tissue engineering approaches hold the promise to fabricate nonthrombogenic biological substitutes for cardiovascular tissues and devices. Endothelial cells (ECs) and stem cells have been used to cover blood-contacting surfaces. Furthermore, for tissue-engineered vascularized tissues and organs, a nonthrombogenic vascular network is essential for mass transfer and the integration of functional tissues and organs into the host upon transplantation. This review discusses the advances in antithrombogenic approaches for surface modifications and cardiovascular tissue engineering. PMID- 21639779 TI - Engineering applications of biomolecular motors. AB - Biomolecular motors, in particular motor proteins from the kinesin and myosin families, can be used to explore engineering applications of molecular motors in general. Their outstanding performance enables the experimental study of hybrid systems, where bio-inspired functions such as sensing, actuation, and transport rely on the nanoscale generation of mechanical force. Scaling laws and theoretical studies demonstrate the optimality of biomolecular motor designs and inform the development of synthetic molecular motors. PMID- 21639780 TI - In vivo delivery of RNAi with lipid-based nanoparticles. AB - RNA interference (RNAi) technology represents a fundamentally new category of treatments for human disease by addressing targets that are traditionally considered undruggable with existing medicines. The major challenge for RNAi based therapy is the delivery system that meets human therapeutic needs. Therefore, engineering vectors with good delivery efficiency and safety profile is an intense area of research. Lipid-based nanoparticles for RNAi have yielded successful advances in vivo and to an extent in clinical trials. In this review, we discuss the barriers in developing lipid-based nanoparticles for in vivo RNAi and different strategies to overcome them. Rational designs that address safety concerns and ensure effective delivery will aid the translation of engineered lipid-based nanoparticles toward the clinic in the foreseeable future. PMID- 21639781 TI - A coevolutionary framework for managing disease-suppressive soils. AB - This review explores a coevolutionary framework for the study and management of disease-suppressive soil microbial communities. Because antagonistic microbial interactions are especially important to disease suppression, conceptual, theoretical, and empirical work on antagonistic coevolution and its relevance to disease suppression is reviewed. In addition, principles of coevolution are used to develop specific predictions regarding the drivers of disease-suppressive potential in soil microbial communities and to highlight important areas for future research. This approach brings an evolutionary perspective to microbial community management and emphasizes the role of species interactions among indigenous nonpathogenic microbes in developing and maintaining disease suppressive activities in soil. PMID- 21639782 TI - Not as they seem. AB - I recount the early influences that directed me toward a career in research and then describe some efforts investigating Cowpea mosaic virus and the satellite RNA of Tobacco ringspot virus. These descriptions have a common theme of surprise, how things often can be not as they are expected to be. Finally, I examine the widely held belief that a plant transgene derived from a distant taxonomic source presents a greater risk than a transgene derived from a closely related plant and contend that this also is a situation in which things may not be as they initially seem. PMID- 21639783 TI - Structure and function of MARTX toxins and other large repetitive RTX proteins. AB - The Repeats-in-Toxins (RTX) family of proteins classically consists of cytolysins and hemolysins. Over the past decade, genome sequencing revealed the existence of very large members of this family. These are all repetitive proteins ranging in size from 200 to 900 kDa that function as toxins or adhesins. Many are exported by Type I secretion. One major new subfamily is the large repetitive RTX adhesins and biofilm-associated proteins. These are characterized by 80- to 300-amino-acid repeats ordered in tandem, although the sequence and number of the repeats vary by protein. The second major new subfamily is the multifunctional-autoprocessing RTX toxins, which are associated with cytotoxicity and pathogenesis. These proteins are in turn distantly related to Yersinia hypothetical RTX proteins that may autoprocess by a similar mechanism. This review discusses current knowledge regarding the structure and function of these new subfamilies of RTX proteins. PMID- 21639784 TI - Fungal protein production: design and production of chimeric proteins. AB - For more than a century, filamentous fungi have been used for the production of a wide variety of endogenous enzymes of industrial interest. More recently, with the use of genetic engineering tools developed for these organisms, this use has expanded for the production of nonnative heterologous proteins. In this review, an overview is given of examples describing the production of a special class of these proteins, namely chimeric proteins. The production of two types of chimeric proteins have been explored: (a) proteins grafted for a specific substrate binding domain and (b) fusion proteins containing two separate enzymatic activities. Various application areas for the use of these chimeric proteins are described. PMID- 21639785 TI - Regulation of alternative sigma factor use. AB - Alternative bacterial sigma factors bind the catalytic core RNA polymerase to confer promoter selectivity on the holoenzyme. The different holoenzymes are thus programmed to recognize the distinct promoter classes in the genome to allow coordinated activation of discrete sets of genes needed for adaptive responses. To form the holoenzymes, the different sigma factors must be available to compete for their common substrate (core RNA polymerase). This review highlights (a) the roles of antisigma factors in controlling the availability of alternative sigma factors and (b) the involvement of diverse regulatory molecules that promote the use of alternative sigma factors through subversion of the domineering housekeeping sigma(70). The latter include the nucleotide alarmone ppGpp and small proteins (DksA, Rsd, and Crl), which directly target the transcriptional machinery to mediate their effects. PMID- 21639786 TI - Life on the outside: the rescue of Coxiella burnetii from its host cell. AB - For over seven decades, Coxiella burnetii, the causative agent of human Q fever, has been considered a prototypical obligate intracellular bacterium that relies exclusively on a eukaryotic cell for growth. Intracellularly, the organism prospers in an acidified, phagolysosome-like vacuole. C. burnetii has evolved to replicate in this harsh compartment by a mechanism involving acid activation of metabolism. The ?2 Mb genome of C. burnetii is about twice the size of genomes of most obligate intracellular bacteria, and the organism's central metabolic pathways are largely intact. The absence of extensive genome reduction suggests the adaptation of C. burnetii to an obligate intracellular lifestyle is a recent evolutionary event. Indeed, insight from early work on C. burnetii metabolism, along with new information gained from metabolic pathway reconstructions, nutrient typing, and expression profiling, allowed the rescue of C. burnetii from its host cell to regain the axenic growth capacity of its ancestors. This advance removes the extensive experimental obstacles associated with intracellular obligatism and opens the door for a renaissance in C. burnetii research. PMID- 21639787 TI - To the happy few. AB - I became a microbiologist in post-WWII Japan, working with Toshio Fukasawa on galactose metabolism in Salmonella. We characterized mutants defective in UDP galactose 4-epimerase, which produced a defective lipopolysaccharide, and this opened doors for me to study lipopolysaccharide biosynthesis, which I pursued in the United States, in the laboratory of Herman Kalckar. After I moved to Berkeley, California, in 1969, I became interested in the function of bacterial outer membranes, a line of work that led to the discovery and characterization of porins as well as the studies on the mycobacterial cell wall. In the early 1990s, it became clear that the outer membrane permeability barrier and the activity of periplasmic beta-lactamase are not enough to explain the resistance of some strains to beta-lactam antibiotics, and the search for the missing factor led to the discovery of RND family multidrug efflux pumps, subjects that continue to fascinate me to this day. PMID- 21639788 TI - Protein quality control in the bacterial periplasm. AB - Protein quality control involves sensing and treatment of defective or incomplete protein structures. Misfolded or mislocalized proteins trigger dedicated signal transduction cascades that upregulate the production of protein quality-control factors. Corresponding proteases and chaperones either degrade or repair damaged proteins, thereby reducing the level of aggregation-prone molecules. Because the periplasm of gram-negative bacteria is particularly exposed to environmental changes and respective protein-folding stresses connected with the presence of detergents, low or high osmolarity of the medium, elevated temperatures, and the host's immune response, fine-tuned protein quality control systems are essential for survival under these unfavorable conditions. This review discusses recent advances in the identification and characterization of the key cellular factors and the emerging general principles of the underlying molecular mechanisms. PMID- 21639789 TI - Eukaryotic picoplankton in surface oceans. AB - The eukaryotic picoplankton is a heterogeneous collection of small protists 1 to 3 ?m in size populating surface oceans at abundances of 10(2) to 10(4) cells ml( 1). Pigmented cells are important primary producers that are at the base of food webs. Colorless cells are mostly bacterivores and play key roles in channeling bacteria to higher trophic levels as well as in nutrient recycling. Mixotrophy and parasitism are relevant but less investigated trophic paths. Molecular surveys of picoeukaryotes have unveiled a large phylogenetic diversity and new lineages, and it is critical to understand the ecological and evolutionary significance of this large and novel diversity. A main goal is to assess how individuals are organized in taxonomic units and how they participate in ecological processes. Picoeukaryotes are convincingly integral members of marine ecosystems in terms of cell abundance, biomass, activity, and diversity and they play crucial roles in food webs and biogeochemical cycles. PMID- 21639790 TI - Regulation of DnaA assembly and activity: taking directions from the genome. AB - To ensure proper timing of chromosome duplication during the cell cycle, bacteria must carefully regulate the activity of initiator protein DnaA and its interactions with the unique replication origin oriC. Although several protein regulators of DnaA are known, recent evidence suggests that DnaA recognition sites, in multiple genomic locations, also play an important role in controlling assembly of pre-replicative complexes. In oriC, closely spaced high- and low affinity recognition sites direct DnaA-DnaA interactions and couple complex assembly to the availability of active DnaA-ATP. Additional recognition sites at loci distant from oriC modulate DnaA-ATP availability by repressing new synthesis, recharging inactive DnaA-ADP, or titrating DnaA. Relying on genomic DnaA binding sites, as well as protein regulators, to control DnaA function appears to provide the best combination of high precision and dynamic regulation necessary to couple DNA replication with cell growth over a range of nutritional conditions. PMID- 21639791 TI - Molecular mechanisms of Staphylococcus aureus iron acquisition. AB - The unique redox potential of iron makes it an ideal cofactor in diverse biochemical reactions. Iron is therefore vital for the growth and proliferation of nearly all organisms, including pathogenic bacteria. Vertebrates sequester excess iron within proteins in order to alleviate toxicity and restrict the amount of free iron available for invading pathogens. Restricting the growth of infectious microorganisms by sequestering essential nutrients is referred to as nutritional immunity. In order to circumvent nutritional immunity, bacterial pathogens have evolved elegant systems that allow for the acquisition of iron during infection. The gram-positive extracellular pathogen Staphylococcus aureus is a commensal organism that can cause severe disease when it gains access to underlying tissues. Iron acquisition is required for S. aureus colonization and subsequent pathogenesis. Herein we review the strategies S. aureus employs to obtain iron through the production of siderophores and the consumption of host heme. PMID- 21639792 TI - Prospects for the future using genomics and proteomics in clinical microbiology. AB - The availability of genome sequences has revolutionized the fields of microbiology and infectious diseases. Indeed, more than 1,000 bacterial genomes and 3,000 viral genomes, including representatives of all significant human pathogens, have been sequenced to date. Owing to this tremendous amount of data, genomes are regarded as chimeras of sequence fragments from various origins. Coupled with novel proteomic analyses, genome sequencing has also resulted in unprecedented advances in pathogen diagnosis and genotyping and in the detection of virulence and antibiotic resistance. Herein, we review current achievements of genomics and proteomics and discuss potential developments for clinical microbiology laboratories. PMID- 21639793 TI - The RpoS-mediated general stress response in Escherichia coli. AB - Under conditions of nutrient deprivation or stress, or as cells enter stationary phase, Escherichia coli and related bacteria increase the accumulation of RpoS, a specialized sigma factor. RpoS-dependent gene expression leads to general stress resistance of cells. During rapid growth, RpoS translation is inhibited and any RpoS protein that is synthesized is rapidly degraded. The complex transition from exponential growth to stationary phase has been partially dissected by analyzing the induction of RpoS after specific stress treatments. Different stress conditions lead to induction of specific sRNAs that stimulate RpoS translation or to induction of small-protein antiadaptors that stabilize the protein. Recent progress has led to a better, but still far from complete, understanding of how stresses lead to RpoS induction and what RpoS-dependent genes help the cell deal with the stress. PMID- 21639794 TI - Unraveling the genetics of cancer: genome sequencing and beyond. AB - Advances in next-generation sequencing technology are enabling the systematic analyses of whole cancer genomes, providing insights into the landscape of somatic mutations and the great genetic heterogeneity that defines the unique signature of an individual tumor. Moreover, integrated studies of the genome, epigenome, and transcriptome reveal mechanisms of tumorigenesis at multiple levels. Progress in sequencing technologies and bioinformatics will improve the costs, sensitivity, and accuracy of detecting somatic mutations, while large scale projects are underway to coordinate cancer genome sequencing at the global level to facilitate the generation and dissemination of high-quality uniform genetic data. These developments will create opportunities for deeper studies of cancer genetics and the clinical application of genome sequencing, and will motivate further research in cancer pathogenesis. PMID- 21639796 TI - Genetics of schizophrenia: new findings and challenges. AB - The work conducted using genome-wide approaches during the past several years has invigorated the field, and represents the dawn of molecular genetics of schizophrenia. The aggregate data increasingly support a combination of rare and common genetic variation in schizophrenia, a major role for polygenic inheritance, and a genetic overlap of schizophrenia and other psychiatric disorders, such as bipolar disorder and autism. The current and upcoming resequencing programs (full exomes to full genomes), in combination with the use of more informative genotyping arrays, will allow a more thorough dissection of the molecular genetics of the disorder. A main challenge for the field is the translation of established genetic associations into a better pathophysiological understanding of schizophrenia. PMID- 21639795 TI - Recent advances in the genetics of Parkinson's disease. AB - Genetic studies have provided valuable insight into the pathological mechanisms underlying Parkinson's disease (PD). The elucidation of genetic components to what was once largely considered a nongenetic disease has given rise to a multitude of cell and animal models enabling the dissection of molecular pathways involved in disease etiology. Here, we review advances obtained from models of dominant mutations in alpha-synuclein and LRRK2 as well as recessive PINK1, parkin and DJ-1 mutations. Recent genome-wide association studies have implicated genetic variability at two of these loci, alpha-synuclein and LRRK2, as significant risk factors for developing sporadic PD. This, coupled with the established role of mitochondrial impairment in both familial and sporadic PD, highlights the likelihood of common mechanisms fundamental to the etiology of both. PMID- 21639797 TI - Putting medical genetics into practice. AB - This article describes a fragment of history on the growing impact of genetics on the practice of medicine over 50 years, as experienced by a medical geneticist who helped to provide services to patients at risk of genetic disorders. It is a personal account influenced by a fascination with chromosomes that has drawn him into many studies, including sex determination, gene discovery, fetal diagnosis, phylogenomics, and karyotype evolution. PMID- 21639798 TI - Control of organelle size: the Golgi complex. AB - The Golgi complex processes secretory proteins and lipids, carries out protein sorting and signaling, and supports growth and composition of the plasma membrane. Golgi complex size likely is regulated to meet the demands of each function, and this may involve differential changes of its distinct subdomains. Nevertheless, the primary size change is elongation of the Golgi ribbon-like network as occurs during Golgi complex doubling for mitosis and during differentiation involving upregulated secretion. One hypothesis states that Golgi complex size is set by the abundance of secretory cargo and Golgi complex components that, through binding vesicle coat complexes, drive vesicle coat formation to alter Golgi complex influx and efflux. Regulation of transport factors controlling Golgi membrane traffic is also observed and may control Golgi complex size, but more work is needed to directly link these events to Golgi complex size regulation, especially during differentiation of specialized cell types. PMID- 21639799 TI - Epithelial progenitor cells in lung development, maintenance, repair, and disease. AB - The vertebrate lung is elegantly patterned to carry out gas exchange and host defense. Similar to other organ systems, endogenous stem and progenitor cells fuel the organogenesis of the lung and maintain homeostasis in the face of normal wear and tear. In the context of acute injury, these progenitor populations are capable of effecting efficient repair. However, chronic injury, inflammation, and immune rejection frequently result in pathological airway remodeling and serious impairment of lung function. Here, we review the development, maintenance, and repair of the vertebrate respiratory system with an emphasis on the roles of epithelial stem and progenitor cells. We discuss what is currently known about their identities, lineage relationships, and the mechanisms that regulate their differentiation along various lineages. A deeper understanding of these progenitor populations will undoubtedly accelerate the discovery of improved cellular, genetic, molecular, and bioengineered therapies for lung disease. PMID- 21639801 TI - Activated NOTCH2 is overexpressed in hepatoblastomas: an immunohistochemical study. AB - Hepatoblastoma is a pediatric malignancy characterized by the uncontrolled proliferation of immature hepatocytes (hepatoblasts). This disease is diagnosed primarily in children younger than 5 years and is disproportionately observed in former premature infants. Cytogenetically, hepatoblastoma is characterized by numerical aberrations, as well as unbalanced translocations involving the proximal region of chromosome 1q. The NOTCH2 gene has been mapped to this locus, and it is well established that the NOTCH gene family is an important regulator of several developmental pathways. Specifically, the NOTCH2 protein is known to delay hepatoblast maturation during early hepatic organogenesis, and the reduction of NOTCH2 expression correlates with the differentiation of hepatoblasts into hepatocytes and biliary cells in the developing liver. We hypothesized that NOTCH2 is involved in the pathogenesis of hepatoblastoma by maintaining a population of undifferentiated hepatoblasts. We studied the immunohistochemical expression of NOTCH2 and its isoforms NOTCH1, NOTCH3, and NOTCH4 and the NOTCH2 primary ligand JAGGED1 in hepatoblastomas. Compared with the normal liver, an increased level of NOTCH2 expression was seen in 22 of 24 (92%) hepatoblastomas. There was no significant staining for other NOTCH isoforms and JAGGED1 in hepatoblastomas. Therefore, we suggest that NOTCH2 expression and activation, independent of JAGGED1 expression, may contribute to the pathogenesis of hepatoblastoma. In the hepatoblastoma sinusoidal vasculature, we saw NOTCH3 and NOTCH1 expression. These observations have potential implications with regard to therapeutic targeting of the NOTCH signaling pathway in hepatoblastomas. PMID- 21639800 TI - Principles of unconventional myosin function and targeting. AB - Unconventional myosins are a superfamily of actin-based motors implicated in diverse cellular processes. In recent years, much progress has been made in describing their biophysical properties, and headway has been made into analyzing their cellular functions. Here, we focus on the principles that guide in vivo motor function and targeting to specific cellular locations. Rather than describe each motor comprehensively, we outline the major themes that emerge from research across the superfamily and use specific examples to illustrate each. In presenting the data in this format, we seek to identify open questions in each field as well as to point out commonalities between them. To advance our understanding of myosins' roles in vivo, clearly we must identify their cellular cargoes and the protein complexes that regulate motor attachment to fully appreciate their functions on the cellular and developmental levels. PMID- 21639803 TI - Cytokines and depression: findings, issues, and treatment implications. AB - Depression has traditionally been classified as a disorder of the brain and CNS, with behavioural manifestations which comprise its symptomatology. That symptomatology includes a range of behaviours that also occur during certain immunological responses to pathogens, and this relationship has engendered a large research literature focussed upon the links between cell messenger cytokines and depression. However, despite many studies of those links, the precise contribution that cytokines make to the development of depression remains unclear. In order to explicate the current state of knowledge of this contribution, a review of literature reviews reported during the last five years, plus a sample of empirical studies reported during the last three years, was conducted. Results indicate that there are many plausible pathways between cytokine responses to pathogens and depression, most notably via 'sickness behaviour', which supports a model of depression as withdrawal behaviour instigated by the brain and that is based upon primitive responses to uncontrollable and life-threatening environmental challenges. However, the precise nature of the mechanisms by which various cytokines communicate with brain regions and influence functions that are trophic to depressive symptomatology remains to be explicated. PMID- 21639804 TI - BDNF-TrkB signalling in fear learning: from genetics to neural networks. AB - Discovering the basic mechanisms in fear encoding and expression is important in many fields, including psychology, sociology, medicine, and neuroscience. Effective treatment for fear-based pathology depends on understanding how fear is learned and regulated. Among the molecular systems required for fear learning and amygdalar synaptic plasticity, brain derived neurtrophic factor (BDNF) and its high affinity receptor Ntrk2/TrkB have been shown to play essential roles. Therefore, we will focus this review on three main aspects; first of all, the impact of Bdnf polymorphism on fear related characteristics in humans and animal models. Secondly, we will discuss BDNF-TrkB activity regulation by epigenetic, transcriptional and post-translational events, and finally we will discuss TrkB BDNF signalling in fear learning. BDNF-TrkB and the signalling activated in this particular form of plasticity are becoming crucial players in fear learning and memory thus highlighting these molecules as potential therapeutic targets in fear related pathologies. PMID- 21639805 TI - Allosteric modulation of ATP-gated P2X receptor channels. AB - Seven mammalian purinergic receptor subunits, denoted P2X1-P2X7, and several spliced forms of these subunits have been cloned. When heterologously expressed, these cDNAs encode ATP-gated non-selective cation channels organized as trimers. All activated receptors produce cell depolarization and promote Ca(2+) influx through their pores and indirectly by activating voltage-gated calcium channels. However, the biophysical and pharmacological properties of these receptors differ considerably, and the majority of these subunits are also capable of forming heterotrimers with other members of the P2X receptor family, which confers further different properties. These channels have three ATP binding domains, presumably located between neighboring subunits, and occupancy of at least two binding sites is needed for their activation. In addition to the orthosteric binding sites for ATP, these receptors have additional allosteric sites that modulate the agonist action at receptors, including sites for trace metals, protons, neurosteroids, reactive oxygen species and phosphoinositides. The allosteric regulation of P2X receptors is frequently receptor-specific and could be a useful tool to identify P2X members in native tissues and their roles in signaling. The focus of this review is on common and receptor-specific allosteric modulation of P2X receptors and the molecular base accounting for allosteric binding sites. PMID- 21639806 TI - Exemestane for breast-cancer prevention in postmenopausal women. AB - BACKGROUND: Tamoxifen and raloxifene have limited patient acceptance for primary prevention of breast cancer. Aromatase inhibitors prevent more contralateral breast cancers and cause fewer side effects than tamoxifen in patients with early stage breast cancer. METHODS: In a randomized, placebo-controlled, double-blind trial of exemestane designed to detect a 65% relative reduction in invasive breast cancer, eligible postmenopausal women 35 years of age or older had at least one of the following risk factors: 60 years of age or older; Gail 5-year risk score greater than 1.66% (chances in 100 of invasive breast cancer developing within 5 years); prior atypical ductal or lobular hyperplasia or lobular carcinoma in situ; or ductal carcinoma in situ with mastectomy. Toxic effects and health-related and menopause-specific qualities of life were measured. RESULTS: A total of 4560 women for whom the median age was 62.5 years and the median Gail risk score was 2.3% were randomly assigned to either exemestane or placebo. At a median follow-up of 35 months, 11 invasive breast cancers were detected in those given exemestane and in 32 of those given placebo, with a 65% relative reduction in the annual incidence of invasive breast cancer (0.19% vs. 0.55%; hazard ratio, 0.35; 95% confidence interval [CI], 0.18 to 0.70; P=0.002). The annual incidence of invasive plus noninvasive (ductal carcinoma in situ) breast cancers was 0.35% on exemestane and 0.77% on placebo (hazard ratio, 0.47; 95% CI, 0.27 to 0.79; P=0.004). Adverse events occurred in 88% of the exemestane group and 85% of the placebo group (P=0.003), with no significant differences between the two groups in terms of skeletal fractures, cardiovascular events, other cancers, or treatment-related deaths. Minimal quality-of-life differences were observed. CONCLUSIONS: Exemestane significantly reduced invasive breast cancers in postmenopausal women who were at moderately increased risk for breast cancer. During a median follow-up period of 3 years, exemestane was associated with no serious toxic effects and only minimal changes in health related quality of life. (Funded by Pfizer and others; NCIC CTG MAP.3 ClinicalTrials.gov number, NCT00083174.). PMID- 21639807 TI - "MAPping" the course of chemoprevention in breast cancer. PMID- 21639808 TI - Improved survival with vemurafenib in melanoma with BRAF V600E mutation. AB - BACKGROUND: Phase 1 and 2 clinical trials of the BRAF kinase inhibitor vemurafenib (PLX4032) have shown response rates of more than 50% in patients with metastatic melanoma with the BRAF V600E mutation. METHODS: We conducted a phase 3 randomized clinical trial comparing vemurafenib with dacarbazine in 675 patients with previously untreated, metastatic melanoma with the BRAF V600E mutation. Patients were randomly assigned to receive either vemurafenib (960 mg orally twice daily) or dacarbazine (1000 mg per square meter of body-surface area intravenously every 3 weeks). Coprimary end points were rates of overall and progression-free survival. Secondary end points included the response rate, response duration, and safety. A final analysis was planned after 196 deaths and an interim analysis after 98 deaths. RESULTS: At 6 months, overall survival was 84% (95% confidence interval [CI], 78 to 89) in the vemurafenib group and 64% (95% CI, 56 to 73) in the dacarbazine group. In the interim analysis for overall survival and final analysis for progression-free survival, vemurafenib was associated with a relative reduction of 63% in the risk of death and of 74% in the risk of either death or disease progression, as compared with dacarbazine (P<0.001 for both comparisons). After review of the interim analysis by an independent data and safety monitoring board, crossover from dacarbazine to vemurafenib was recommended. Response rates were 48% for vemurafenib and 5% for dacarbazine. Common adverse events associated with vemurafenib were arthralgia, rash, fatigue, alopecia, keratoacanthoma or squamous-cell carcinoma, photosensitivity, nausea, and diarrhea; 38% of patients required dose modification because of toxic effects. CONCLUSIONS: Vemurafenib produced improved rates of overall and progression-free survival in patients with previously untreated melanoma with the BRAF V600E mutation. (Funded by Hoffmann-La Roche; BRIM-3 ClinicalTrials.gov number, NCT01006980.). PMID- 21639809 TI - Been there, not done that--melanoma in the age of molecular therapy. PMID- 21639810 TI - Ipilimumab plus dacarbazine for previously untreated metastatic melanoma. AB - BACKGROUND: Ipilimumab monotherapy (at a dose of 3 mg per kilogram of body weight), as compared with glycoprotein 100, improved overall survival in a phase 3 study involving patients with previously treated metastatic melanoma. We conducted a phase 3 study of ipilimumab (10 mg per kilogram) plus dacarbazine in patients with previously untreated metastatic melanoma. METHODS: We randomly assigned 502 patients with previously untreated metastatic melanoma, in a 1:1 ratio, to ipilimumab (10 mg per kilogram) plus dacarbazine (850 mg per square meter of body-surface area) or dacarbazine (850 mg per square meter) plus placebo, given at weeks 1, 4, 7, and 10, followed by dacarbazine alone every 3 weeks through week 22. Patients with stable disease or an objective response and no dose-limiting toxic effects received ipilimumab or placebo every 12 weeks thereafter as maintenance therapy. The primary end point was overall survival. RESULTS: Overall survival was significantly longer in the group receiving ipilimumab plus dacarbazine than in the group receiving dacarbazine plus placebo (11.2 months vs. 9.1 months, with higher survival rates in the ipilimumab dacarbazine group at 1 year (47.3% vs. 36.3%), 2 years (28.5% vs. 17.9%), and 3 years (20.8% vs. 12.2%) (hazard ratio for death, 0.72; P<0.001). Grade 3 or 4 adverse events occurred in 56.3% of patients treated with ipilimumab plus dacarbazine, as compared with 27.5% treated with dacarbazine and placebo (P<0.001). No drug-related deaths or gastrointestinal perforations occurred in the ipilimumab-dacarbazine group. CONCLUSIONS: Ipilimumab (at a dose of 10 mg per kilogram) in combination with dacarbazine, as compared with dacarbazine plus placebo, improved overall survival in patients with previously untreated metastatic melanoma. The types of adverse events were consistent with those seen in prior studies of ipilimumab; however, the rates of elevated liver-function values were higher and the rates of gastrointestinal events were lower than expected on the basis of prior studies. (Funded by Bristol-Myers Squibb; ClinicalTrials.gov number, NCT00324155.). PMID- 21639812 TI - Transcatheter aortic-valve implantation--at what price? PMID- 21639813 TI - Review of approved pioglitazone combinations for type 2 diabetes. AB - INTRODUCTION: Pioglitazone is approved in combination with several other blood glucose-lowering drugs for the treatment of type 2 diabetes mellitus (T2DM). Beyond lowering blood glucose levels, each combination of different blood-glucose lowering drugs for the treatment of T2DM evolves specific pleiotropic effects, which might be considered on an individual basis in a certain patient. AREAS COVERED: The objective of this article is to provide a short review of the pathophysiology of T2DM and to provide a rationale for the combination of pioglitazone with other antidiabetic drugs, based on a pathophysiological understanding of T2DM. Therefore, a PubMed search was undertaken covering the search terms 'pioglitazone', 'antidiabetic drugs' and 'combination therapy'. EXPERT OPINION: Treatment with pioglitazone in T2DM was shown to improve insulin resistance and blood glucose levels without increasing the risk of hypoglycemia. Beyond those metabolic activities, pioglitazone was shown to evolve anti inflammatory and anti-atherogenic effects. It seems useful to combine different antidiabetic drugs based on the specific needs and contraindications in an individual patient. The treating of T2DM patients by addressing not only glucose control, but also the underlying pathophysiological etiology might help to improve patient prognosis in the long run, especially with regard to the vascular complications. PMID- 21639814 TI - Expert opinion on current therapies for nonalcoholic fatty liver disease. AB - INTRODUCTION: Nonalcoholic fatty liver disease (NAFLD) is rapidly becoming one of most common liver diseases in industrialized countries owing to the increasing prevalence of obesity and being overweight. Until now, loss of weight and physical activity have represented the cornerstone of treatment, but they are very difficult to achieve and to maintain. Therefore, new treatments based on pathogenetic mechanisms leading to NAFLD are under evaluation to establish an effective pharmacological therapy for this disorder. AREAS COVERED: An overview of current therapeutic interventions to treat NAFLD is given. This review provides evidence of the efficacy of natural and pharmacological agents used so far in the treatment of both adult and pediatric NAFLD, on the basis of clinical trials published in the last 10 years. EXPERT OPINION: In the last 10 years, many pharmacological agents on the basis of the pathogenetic mechanism of NAFLD have been attempted, but so far guidelines for the management of NAFLD are lacking. We believe that the advance in the understanding of pathogenesis and factors involved in the progression of the disease may disclose the way to defining new, solid, therapeutic strategies. A multidisciplinary approach considering the risk factors and comorbidities of fatty liver will represent in the future a successful therapeutic strategy for NAFLD. PMID- 21639811 TI - Transcatheter versus surgical aortic-valve replacement in high-risk patients. AB - BACKGROUND: The use of transcatheter aortic-valve replacement has been shown to reduce mortality among high-risk patients with aortic stenosis who are not candidates for surgical replacement. However, the two procedures have not been compared in a randomized trial involving high-risk patients who are still candidates for surgical replacement. METHODS: At 25 centers, we randomly assigned 699 high-risk patients with severe aortic stenosis to undergo either transcatheter aortic-valve replacement with a balloon-expandable bovine pericardial valve (either a transfemoral or a transapical approach) or surgical replacement. The primary end point was death from any cause at 1 year. The primary hypothesis was that transcatheter replacement is not inferior to surgical replacement. RESULTS: The rates of death from any cause were 3.4% in the transcatheter group and 6.5% in the surgical group at 30 days (P=0.07) and 24.2% and 26.8%, respectively, at 1 year (P=0.44), a reduction of 2.6 percentage points in the transcatheter group (upper limit of the 95% confidence interval, 3.0 percentage points; predefined margin, 7.5 percentage points; P=0.001 for noninferiority). The rates of major stroke were 3.8% in the transcatheter group and 2.1% in the surgical group at 30 days (P=0.20) and 5.1% and 2.4%, respectively, at 1 year (P=0.07). At 30 days, major vascular complications were significantly more frequent with transcatheter replacement (11.0% vs. 3.2%, P<0.001); adverse events that were more frequent after surgical replacement included major bleeding (9.3% vs. 19.5%, P<0.001) and new-onset atrial fibrillation (8.6% vs. 16.0%, P=0.006). More patients undergoing transcatheter replacement had an improvement in symptoms at 30 days, but by 1 year, there was not a significant between-group difference. CONCLUSIONS: In high-risk patients with severe aortic stenosis, transcatheter and surgical procedures for aortic valve replacement were associated with similar rates of survival at 1 year, although there were important differences in periprocedural risks. (Funded by Edwards Lifesciences; Clinical Trials.gov number, NCT00530894.). PMID- 21639815 TI - Early changes in parathyroid hormone concentrations in HIV-infected patients initiating antiretroviral therapy with tenofovir. AB - Initiation of combined antiretroviral therapy (cART) is associated with bone loss, which may be more intense with regimens including tenofovir. The underlying mechanisms are not well understood. Cross-sectional data have linked tenofovir with higher parathyroid hormone (PTH) concentrations in patients with vitamin D deficiency. We performed a longitudinal study with a 48-week follow-up to evaluate sequential changes in PTH and 25-hydroxyvitamin D [25(OH)D] levels in patients starting cART with either tenofovir/emtricitabine or abacavir/lamivudine. Fifty-seven patients were included, 31 initiating tenofovir/emtricitabine and 26 initiating abacavir/lamivudine. Median PTH levels turned out to be significantly higher among tenofovir/emtricitabine users at week 4 (p=0.01), week 24 (p=0.008), and week 36 (p=0.02), and were above the upper limits of normal values (ULN) at weeks 24, 36, and 48 only in patients receiving tenofovir/emtricitabine. 25(OH)D, serum and urine calcium and phosphate, and renal-tubular maximum reabsorption of phosphate to the glomerular filtration rate (TmP/GFR) levels did not differ between the two treatment arms over the study period. Among tenofovir/emtricitabine users, median (interquartile range) PTH concentrations were significantly higher in patients with suboptimal 25(OH)D levels (<30 MUg/liter) at week 24 [63 (57.8-82.4) ng/liter vs. 54.3 (34.4 63.067.5) ng/liter, p=0.05] and week 48 [67.5 (59.6-86.0) ng/liter vs. 41.9 (37.3 68.8) ng/liter, p=0.03]. A multivariable logistic regression model showed that tenofovir/emtricitabine use was an independent predictor of high PTH levels (>=53 ng/liter). Starting cART with tenofovir regimens is associated with an elevation in PTH plasma concentrations soon after introducing the drug. Suboptimal baseline 25(OH)D levels increase the risk of developing secondary hyperparathyroidism among tenofovir users. PMID- 21639816 TI - Pharmacokinetic evaluation of levocetirizine. AB - INTRODUCTION: There have recently been guidelines developed for the diagnosis and treatment of rhinitis and urticaria. For both conditions, second-generation antihistamines remain as the first-line therapy. AREAS COVERED: The article presents the current pharmacology, chemical properties, pharmacokinetics and metabolism of levocetirizine. The article also reviews the clinical efficacy of levocetirizine for seasonal allergic and perennial rhinitis, as well as chronic urticaria. The article is formed through the review of all the published literature in English retrieved from the PubMed/MEDLINE database between 1966 and March 2011 using the search terms: levocetirizine, allergic rhinitis, chronic urticaria and antihistamine. Furthermore, the article also reviews data provided by the manufacturer in addition to reports from governmental agencies. EXPERT OPINION: Levocetirizine has several pharmacokinetic properties that are desirable for an antihistamine providing a combination of both potency and safety. Its clinical advantages are derived from its rapid and extensive absorption, limited distribution and its very low degree of metabolism. Furthermore, levocetirizine scores very highly in terms of clinical efficacy as well as in patient/physician satisfaction studies. Given the lack of large multi-center studies that compare the treatment options for urticaria, clinicians must rely on potency studies when choosing treatment and levocetirizine does score very highly. However, other potent skin antihistamines, such as desloratadine or fexofenadine, should be preferred for patients who have a strict contraindication to the sedative effects of the drug. PMID- 21639817 TI - Review of fesoterodine. AB - INTRODUCTION: Overactive bladder syndrome is a common condition that adversely affects the quality of life. It is mainly treated with a combination of bladder retraining and antimuscarinics. In a quest to reduce the side effect profile of these drugs, whilst improving their efficacy, more bladder-selective antimuscarinics were developed. One of the more recent of these antimuscarinics which has come to the market is fesoterodine. This review examines the evidence of the safety and efficacy of this drug. AREAS COVERED: A literature search performed identified two main multi-center trials which highlight the safety, efficacy and tolerability of fesoterodine. These together with the pharmacologic properties of the drug are discussed at length throughout the review. An expert opinion is then formulated based on the current evidence available and on comparison with other antimuscarinics. EXPERT OPINION: It is concluded that fesoterodine has the added advantage of flexible dosing over some other antimuscarinics. It does, however, have a similar tolerability and side effect profile to other antimuscarinics and is, therefore, unlikely to revolutionize the treatment of the overactive bladder. PMID- 21639818 TI - A high-fat diet temporarily accelerates gastrointestinal transit and reduces satiety in men. AB - High-fat (HF) diets of 2 weeks have been shown to accelerate gastrointestinal (GI) transit and decrease satiety. However, the effects of HF diets on GI transit over longer periods than 2 weeks are unknown. We hypothesize that over 4 weeks, GI transit of a HF test meal will accelerate. The study was a repeated measures design with 10 male volunteers completing a 1-week HF diet intervention and 7 completing a 4-week HF diet intervention with testing once a week on the same day throughout the 4 weeks. Gastric emptying (GE) was measured using the (13)C octanoic acid breath test and mouth-to-caecum transit time (MCTT) using the inulin H(2) breath test. Satiety was analysed using visual analogue scales and an ad libitum buffet meal. Body mass increased by 1.3 kg over the 4 weeks (p = 0.036). GE latency time decreased from 45 +/- 8 to 41 +/- 10 min (p = 0.047) over 1 week but there were no changes in any GE parameters over the 4 weeks. MCTT was accelerated over 1 week (p = 0.036) from 308 +/- 43 to 248 +/- 83 min. However, over the 4-week period, there was no change. Volunteers became more hungry and desire to eat became greater after 1 week (p = 0.01). Changes in satiety were also evident over the 4 weeks. Satiety was reduced in the primary weeks and then returned to baseline towards the end of the intervention. GI adaptation to a HF diet occurred over a 1-week period and returned to pre-diet levels at the end of 4 weeks. PMID- 21639819 TI - Microwave-assisted hydrolysis of lutein and zeaxanthin esters in marigold (Tagetes erecta L.). AB - Saponification of lutein and zeaxanthin was performed by microwave-assisted hydrolysis (MAH) and analysed by ultra performance liquid chromatography. The optimal condition of MAH was studied, and the degradation or isomerization of lutein and zeaxanthin were estimated under MAH. The concentrations of lutein and zeaxanthin in 20 marigold samples were assessed by saponification using traditional heater and MAH, the regression coefficient of lutein obtained by two methods was 0.9688 and that of zeaxanthin was 0.9527. The limit of detection for lutein and zeaxanthin was 0.05 and 0.1 MUg/ml, respectively, and the limit of quantification for lutein and zeaxanthin was 0.05 mg/100 g and 0.1 mg/100 g, respectively. PMID- 21639820 TI - Impact of apple and grape juice consumption on the antioxidant status in healthy subjects. AB - It is reported that consumption of antioxidant-rich foods significantly increased plasma total antioxidant capacity (T-AOC) in humans. Also, it is proved that the antioxidants from plant foods improve the body's antioxidant defence by acting additively and synergistically. As a result, rational combination of antioxidant rich foods is recommended to population in the prevention of oxidative stress related diseases. Both apple and grape are antioxidant-rich fruits. In this study, 2 weeks dietary intervention study was carried out in 25 healthy subjects to investigate the influences of apple and grape juices consumption on body antioxidant status. Our results indicated that 2 weeks of apple and grape juice consumption increased the plasma T-AOC and decreased the concentration of malondialdehyde. However, no change was found in the content of plasma carbonyl. Erythrocyte glutathione peroxidase and catalase activities were enhanced by 2 weeks of fruit juice consumption; however, no change was found in the activity of erythrocyte superoxide dismutase. The in vitro comet assay results indicated that apple and grape juice consumption did not influence lymphocyte damage upon hydrogen peroxide treatment. Urinary 8-hydroxy-2-deoxyguanosine content was not affected by 2 weeks of fruit juice intervention. These findings indicated that concomitant intake of apple and grape juice was efficient in enhancing the body's antioxidant status. PMID- 21639821 TI - Enzastaurin hydrochloride for lymphoma: reassessing the results of clinical trials in light of recent advances in the biology of B-cell malignancies. AB - INTRODUCTION: The B-cell receptor (BCR) is critical for the development and persistence of B-cell non-Hodgkin lymphoma (B-NHL). Protein kinase C-beta (PKC-?) has been identified as one of the key signaling hubs downstream of the BCR and constitutes a valuable target in B-NHL. As a potent PKC-? inhibitor, enzastaurin is currently being tested in Phase II/III trials. AREAS COVERED: This review summarizes the latest results and ongoing clinical trials with enzastaurin in light of basic scientific advances in the understanding of various lymphoid cancers, including diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma (MCL), follicular lymphoma (FL), chronic lymphocytic leukemia (CLL) and Waldenstr?m's macroglobulinemia (WM). EXPERT OPINION: While its continued clinical development is uncertain, enzastaurin should be regarded as a stepping stone for the development of future therapies; indeed, the recent research has provided valuable insight into the possible molecular mechanisms that explain its limited clinical activity especially in the treatment of DLBCL and MCL. It should be noted that there is still some interest in enzastaurin, in combination, for the treatment of WM. PMID- 21639823 TI - Modification of the VerifyNow(r) P2Y12 test BASE channel to accommodate high levels of P2Y(12) antagonism. AB - The VerifyNow(r) P2Y12 (VN-P2Y12) test reports thienopyridine-mediated platelet inhibition relative to a "BASE" channel, potentially eliminating the need for predrug patient assessment, by activating platelets through a P2Y(12)-independent pathway. The original formulation of the BASE channel used a protease activated receptor-1 (PAR-1) peptide as agonist. However, more potent P2Y(12) antagonism required more complete activation of platelet thrombin receptors for the BASE measurement in order to negate any contribution of the P2Y(12) receptor. Accordingly, the current BASE channel formulation consists of both PAR-1 and protease activated receptor-4 (PAR-4) activating peptides to facilitate a higher degree of platelet activation. The aim of this study was to compare the performance of PAR-1 versus PAR-1/PAR-4 activating peptides as the BASE channel formulation using prasugrel's active metabolite, R-138727, in vitro to achieve high-grade P2Y(12) inhibition. Blood samples from 20 healthy donors were spiked in vitro with R-138727 at concentrations that include plasma levels achieved following prasugrel administration and were incubated for 30 minutes at 37 degrees C. All samples were run in triplicate using both the PAR-1 and the PAR 1/PAR-4 BASE formulation in the VN-P2Y12 test device. The data confirmed the sensitivity of the original BASE formulation to high-grade P2Y(12) inhibition as reflected in the concentration-dependent decrease in values. Incorporation of PAR 4 activating peptide eliminated the effect of P2Y(12) blockade at all concentrations of R-138727. Thus, the use of PAR-1/PAR-4 in the BASE channel of the VN-P2Y12 cartridge addresses the impact of high grade P2Y(12) blockade and may allow more accurate reporting of "% inhibition" in patients treated with more effective P2Y(12) antagonists. PMID- 21639824 TI - Cardiac AA amyloidosis in a patient with rheumatoid arthritis and systemic sclerosis: the therapeutic potential of biological reagents. PMID- 21639825 TI - Oral sildenafil in skin ulcers secondary to systemic sclerosis. PMID- 21639826 TI - Contribution of TNF-alpha-308 gene polymorphisms to susceptibility to the development of uveitis in Korean patients with HLA-B27-positive ankylosing spondylitis. PMID- 21639827 TI - Adult-onset malignancy-associated Henoch-Schonlein purpura. PMID- 21639828 TI - CYB5D2 enhances HeLa cells survival of etoposide-induced cytotoxicity. AB - Cytochrome b5 domain containing 2 (CYB5D2) (neuferricin) belongs to the family of membrane-associated progesterone receptors (MAPRs). MAPRs affect multiple cellular processes, including proliferation, differentiation, and survival. Consistent with these observations, we report here that CYB5D2 enhances HeLa cells survival of etoposide (ETOP)-mediated cytotoxicity. Overexpression of CYB5D2 enhanced the survival of HeLa cells compared with HeLa cells transfected with empty vector (EV) upon ETOP treatment. As ETOP initiates ATM-dependent DNA damage response (DDR), we were able to show that CYB5D2 did not affect ETOP induced DDR. In line with these observations, CYB5D2 did not protect HeLa cells from UV-induced cytotoxicity. Additionally, CYB5D2 had no effects on TNFalpha induced apoptosis. Collectively, CYB5D2 enhances HeLa cell survival of ETOP induced cytotoxicity with some specificity. CYB5D2 contains a cytochrome b5 (cyt b5) domain and a transmembrane (TM) motif. Both domains are required for CYB5D2 mediated protection of HeLa cells from ETOP-induced cytotoxicity. In an effort to search for the underlying mechanisms, we have profiled gene expression between HeLa-CYB5D2 and HeLa-EV cells. Although ectopic CYB5D2 does not massively alter gene expression, the expression of several transcripts was affected more than 2 fold, suggesting that they may contribute to CYB5D2-mediated HeLa cell survival of ETOP treatment. PMID- 21639829 TI - The lipopolysaccharide core of Actinobacillus suis and its relationship to those of Actinobacillus pleuropneumoniae. AB - The Gram-negative bacteria Actinobacillus suis colonizes the upper respiratory and genital tracts of swine. Along with capsular polysaccharides, lipopolysaccharides (O-chain->core->lipid A~cell) are a main cell-surface component of A. suis. In this study, we determined that A. suis lipopolysaccharide incorporates a conserved core that shares some structural features with several core types of A. pleuropneumoniae . These common core structural features likely account for the observed serological cross-reactivity between A. suis and A. pleuropneumoniae, and the data suggest that the structural epitopes responsible for immunogenicity are those in the outer core domain. PMID- 21639830 TI - The isomerase Rrd1 mediates rapid loss of the Sgs1 helicase in response to rapamycin. AB - In Saccharomyces cerevisiae , rapamycin exposure inhibits the target of rapamycin (TOR) signaling pathway, causing a profound alteration in the transcription pattern of many genes, including those involved in ribosome biogenesis and nutritional changes. Deletion of the RRD1 gene encoding a peptidyl prolyl isomerase resulted in mutants that are resistant to rapamycin. These rrd1Delta mutants are unable to efficiently downregulate genes such as ribosomal protein genes, or to upregulate genes involved in diauxic shift. It is believed that the isomerase function of Rrd1 plays a role in changing the transcriptional profile upon rapamycin exposure. Herein, we set out to search for genes that when deleted in the rrd1Delta mutant would suppress the rapamycin-resistant phenotype. The analysis revealed that deletion of the SGS1 gene in the rrd1Delta mutant partially suppresses the rapamycin-resistant phenotype of the single rrd1Delta mutant. SGS1 encodes a helicase that functions in many biological processes, including transcriptional regulation. We further show, and for the first time, that Sgs1 is rapidly lost in the parent cells in response to rapamycin, but not by other agents. Interestingly, Sgs1 reduction was completely blocked in the rrd1Delta mutant, suggesting that Rrd1 is required to mediate this process. Genes such as PUT4 and HSP42, known to be upregulated in the parent in response to rapamycin, were not induced in the rrd1Delta mutant if the SGS1 gene was deleted. Since Sgs1 plays a role in transcriptional regulation, we propose that it acts as a repressor of a subset of rapamycin responsive genes. Thus, the observed Rrd1 dependent reduction in Sgs1 level may promote expression of specific classes of genes in response to rapamycin. PMID- 21639831 TI - Fluorescence correlation spectroscopy in biology, chemistry, and medicine. AB - This review describes the method of fluorescence correlation spectroscopy (FCS) and its applications. FCS is used for investigating processes associated with changes in the mobility of molecules and complexes and allows researchers to study aggregation of particles, binding of fluorescent molecules with supramolecular complexes, lipid vesicles, etc. The size of objects under study varies from a few angstroms for dye molecules to hundreds of nanometers for nanoparticles. The described applications of FCS comprise various fields from simple chemical systems of solution/micelle to sophisticated regulations on the level of living cells. Both the methodical bases and the theoretical principles of FCS are simple and available. The present review is concentrated preferentially on FCS applications for studies on artificial and natural membranes. At present, in contrast to the related approach of dynamic light scattering, FCS is poorly known in Russia, although it is widely employed in laboratories of other countries. The goal of this review is to promote the development of FCS in Russia so that this technique could occupy the position it deserves in modern Russian science. PMID- 21639832 TI - Biological chemistry as a foundation of DNA genealogy: the emergence of "molecular history". AB - This paper presents the basis of DNA genealogy, a new field of science, which is currently emerging as an unusual blend of biochemistry, history, linguistics, and chemical kinetics. The methodology of the new approach is comprised of chemical (biological) kinetics applied to a pattern of mutations in non-recombinant fragments of DNA (Y chromosome and mtDNA, the latter not being considered in this overview). The goal of the analysis is to translate DNA mutation patterns into time spans to the most recent common ancestors of a given population or tribe and to the dating of ancient migration routes. To illustrate this approach, time spans to the common ancestors are calculated for ethnic Russians, that is Eastern Slavs (R1a1 tribe), Western Slavs (I1 and I2 tribes), and Northern (or Uralic) Slavs (N1c tribe), which were found to live around 4600 years before present (R1a1), 3650 ybp (I1), 3000 and 10,500 ybp (I2, two principal DNA lineages), and 3525 ybp (N1c) (confidence intervals are given in the main text). The data were compared with the respective dates for the nearest common ancestor of the R1a1 "Indo-European" population in India, who lived 4050 years before present, whose descendants represent the majority of the upper castes in India today (up to 72%). Furthermore, it was found that the haplotypes of ethnic Russians of the R1a1 haplogroup (up to 62% of the population in the Russian Federation) and those of the R1a1 Indians (more than 100 million today) are practically identical to each other, up to 67-marker haplotypes. This essentially solves a 200-year-old mystery of who were the Aryans who arrived in India around 3500 years before the present. Haplotypes and time spans to the ancient common ancestors were also compared for the ethnic Russians of haplogroups I1 and I2, on one hand, and the respective I1 and I2 populations in Eastern and Western Europe and Scandinavia, on the other. It is suggested that the approach described in this overview lays the foundation for "molecular history", in which the principal tool is high technology analysis of DNA molecules of both our contemporaries and excavated ancient DNA samples, along with their biological kinetics. PMID- 21639833 TI - IgM and its receptors: structural and functional aspects. AB - This review combines the data obtained before the beginning of the 1990s with results published during the last two decades. The predominant form of the IgM molecule is a closed ring composed of five 7S subunits and a J chain. The new model of spatial structure of the pentamer postulates nonplanar mushroom-shaped form of the molecule with the plane formed by a radially-directed Fab regions and central protruding portion consisting of Cu4 domains. Up to the year 2000 the only known Fc-receptor for IgM was pIgR. Interaction of IgM with pIgR results in secretory IgM formation, whose functions are poorly studied. The receptor designated as Fcalpha/uR is able to bind IgM and IgA. It is expressed on lymphocytes, follicular dendritic cells, and macrophages. A receptor binding IgM only named FcuR has also been described. It is expressed on T- and B-lymphocytes. The discovery of new Fc-receptors for IgM requires revision of notions that interactions between humoral reactions involving IgM and the cells of the immune system are mediated exclusively by complement receptors. In the whole organism, apart from IgM induced by immunization, natural antibodies (NA) are present and comprise in adults a considerable part of the circulating IgM. NA are polyreactive, germ-line-encoded, and emerge during embryogenesis without apparent antigenic stimuli. They demonstrate a broad spectrum of antibacterial activity and serve as first line of defense against microbial and viral infections. NA may be regarded as a transitional molecular form from invariable receptors of innate immunity to highly diverse receptors of adaptive immunity. By means of interaction with autoantigens, NA participate in maintenance of immunological tolerance and in clearance of dying cells. At the same time, NA may act as a pathogenic factor in atherosclerotic lesion formation and in development of tissue damage due to ischemia/reperfusion. PMID- 21639834 TI - Promyelocytic leukemia protein interacts with werner syndrome helicase and regulates double-strand break repair in gamma-irradiation-induced DNA damage responses. AB - We show here that gamma-irradiation leads to the translocation of endogenous Werner syndrome helicase (WRN) from nucleoli to nucleoplasmic DNA double strand breaks (DSBs), and WRN plays a role in damage repair. The relocation of WRN after irradiation was perturbed by promyelocytic leukemia protein (PML) knockdown and enhanced by PML IV overexpression. PML IV physically interacted with WRN after irradiation. Amino acids (a.a.) 394 to 433 of PML were necessary for this interaction and the nucleoplasmic translocation of WRN and were involved in DSB repair and cellular sensitivity to gamma-irradiation. Taken together, our results provide molecular support for a model in which PML IV physically interacts with and regulates the translocation of WRN for DNA damage repair through its 394-433 a.a. domain. PMID- 21639835 TI - Apomyoglobin mutants with single point mutations at val10 can form amyloid structures at permissive temperature. AB - Formation of amyloid-like protein aggregates in human organs and tissues underlies many serious diseases, therefore being in the focus of numerous biochemical, medical, and molecular biological studies. So far, formation of amyloids by globular proteins has been studied mostly under conditions that strongly destabilized their native structure. Here we present our results obtained at permissive temperature by thioflavin T fluorescence, far UV CD, IR spectroscopy, and electron microscopy. We used apomyoglobin and its mutants with Ala or Phe substituted for Val10 that are structurally close to wild type apomyoglobin. It is shown that at permissive temperature the ability of the protein to form amyloids depends on the extent of its structural destabilization, but not on hydrophobicity of the substituting residue. A possible difference between amyloids formed by strongly destabilized proteins and those yielded by proteins with a slightly fluctuating native structure, as well as the stroke and infarction effect on the ability of proteins to form amyloid structures, are discussed. PMID- 21639836 TI - Three-dimensional structures of noncovalent complexes of Citrobacter freundii methionine gamma-lyase with substrates. AB - Crystal structures of Citrobacter freundii methionine gamma-lyase complexes with the substrates of gamma- (L-1-amino-3-methylthiopropylphosphinic acid) and beta- (S-ethyl-L-cysteine) elimination reactions and the competitive inhibitor L norleucine have been determined at 1.45, 1.8, and 1.63 A resolution, respectively. All three amino acids occupy the active site of the enzyme but do not form a covalent bond with pyridoxal 5'-phosphate. Hydrophobic interactions between the active site residues and the side groups of the substrates and the inhibitor are supposed to cause noncovalent binding. Arg374 and Ser339 are involved in the binding of carboxyl groups of the substrates and the inhibitor. The hydroxyl of Tyr113 is a potential acceptor of a proton from the amino groups of the amino acids. PMID- 21639837 TI - Protective effect of L-Arginine administration on proteins of unloaded m. soleus. AB - Cytoskeletal and contractile proteins degenerate during functional unloading of muscle. The ratio of myosin heavy chain (MHC) expression changes simultaneously. We have supposed that NO can be a signal molecule related to the regulation of protein metabolism upon muscle unloading. To test this hypothesis, Wistar rats underwent functional unloading for 14 days without and with peroral administration of L-arginine (500 mg/kg) as NO precursor. Significant decreases in m. soleus mass, NO, nNOS, dystrophin, Hsp90, p-S6K, and type I MHC mRNA contents were found in the group of animals with unloading without preparation compared to those in control and in the group with unloading and administration of L-arginine; at the same time, increased contents of atrogin-1/MAFbx and MuRF-1 (p < 0.05) were found. No difference in the IGF-1 mRNA content between all three groups was found. Atrophy was significantly less pronounced in the group with unloading and L-arginine administration compared to that without the amino acid, and no destruction of cytoskeletal proteins was observed. We conclude that administration of L-arginine upon functional unloading decreases the extent of m. soleus atrophy, prevents the decrease in it of type I MHC mRNA, and blocks destructive changes in some cytoskeletal proteins. Such effect can be due to the absence of increase in this group of the content of some ubiquitin ligases and decreased intensity of the p70S6 kinase synthesis marker. PMID- 21639838 TI - Complex of digestive proteinases of Galleria mellonella Caterpillars: composition, properties, and limited proteolysis of Bacillus thuringiensis endotoxins. AB - The complex of digestive proteinases in caterpillars of the greater wax moth Galleria mellonella was studied. Using chromogenic substrates and inhibitor analysis, it was found that serine proteinases play a key role in this complex. Three anionic and two cationic forms of trypsin and one anionic and one cationic form of chymotrypsin were identified by zymography in the midgut extract of G. mellonella. The most active trypsin was purified to electrophoretic homogeneity, and its N-terminal amino acid sequence was shown to be identical to that of mature trypsin from Plodia interpunctella. Midgut extract from G. mellonella was capable of processing Cry-proteins from Bacillus thuringiensis ssp. galleriae. Enzymes with tryptic and chymotryptic activities participate in this process, and activation of protoxin Cry9A is not the rate-limiting stage in the toxic action of this protein on the greater wax moth. PMID- 21639839 TI - Kinetics of chaperone activity of proteins Hsp70 and Hdj1 in human leukemia u-937 cells after preconditioning with thermal shock or compound u-133. AB - Kinetics of the chaperone activity of proteins Hsp70 and Hdj1 were analyzed in human U-937 promonocytes during their response to heat shock or to treatment with the echinochrome triacetyl glucoside derivative U-133. To measure the chaperone activity of both proteins, a special test was developed for their recognition and binding of a denatured protein. Using this test, the chaperone activity could be concurrently estimated in large numbers of cellular or tissue extracts. We also estimated the contents of both chaperones in cells by immunoblotting. The values for contents of Hsp70 and Hdj1 obtained by two independent test systems coincided, and this suggested that the substrate-binding activity could change proportionally to the chaperone content in the protein mixture. Therefore, the test developed by us can be employed for high throughput screening of drugs activating cellular chaperones. The analysis of quantity and activity of two cellular chaperones during the cell response to heat stress or to the drug-like substance U-133 showed that both factors caused the accumulation of chaperones with similar kinetics. We conclude that the efficiency of drug preconditioning could be close to the efficiency of hyperthermia and that the high activity of chaperones could be retained in human cells for no less than 1.5 days. PMID- 21639840 TI - Immunostimulating effect of the synthetic peptide octarphin corresponding to beta endorphin fragment 12-19. AB - We have synthesized the peptide TPLVTLFK corresponding to beta-endorphin fragment 12-19 (dubbed octarphin) and its analogs (LPLVTLFK, TLLVTLFK, TPLVLLFK, TPLVTLLK, TPLVTLFL). The octarphin peptide was labeled with tritium (specific activity 28 Ci/mol), and its binding to murine peritoneal macrophages was studied. [3H]Octarphin was found to bind to macrophages with high affinity (K(d) = 2.3 +/- 0.2 nM) and specificity. The specific binding of [3H]octarphin was inhibited by unlabeled b-endorphin and the selective agonist of nonopioid b-endorphin receptor synthetic peptide immunorphin (SLTCLVKGFY) (K(i) = 2.7 +/- 0.2 and 2.4 +/- 0.2 nM, respectively) and was not inhibited by unlabeled naloxone, a-endorphin, gamma endorphin, or [Met(5)]enkephalin (K(i) > 10 mM). Inhibitory activity of unlabeled octarphin analogs was more than 100 times lower than that of unlabeled octarphin. Octarphin was shown to stimulate activity of murine immunocompetent cells in vitro and in vivo: at concentration of 1-10 nM it enhanced the adhesion and spreading of peritoneal macrophages as well as their ability to digest bacteria of Salmonella typhimurium virulent strain 415 in vitro; the peptide administered intraperitoneally at a dose of 20 ug/animal on day 7, 3, and 1 prior to isolation of cells increased activity of peritoneal macrophages as well as spleen T- and B lymphocytes. PMID- 21639841 TI - Structural-functional features of plant isoperoxidases. AB - Current data on structural--functional features of plant peroxidases and their involvement in functioning of the pro-/antioxidant system responding to stress factors, especially those of biotic origin, are analyzed. The collection of specific features of individual isoforms allows a plant to withstand an aggressive influence of the environment. Expression of some genes encoding different isoperoxidases is regulated by pathogens (and their metabolites), elicitors, and hormone-like compounds; specific features of this regulation are considered in detail. It is suggested that isoperoxidases interacting with polysaccharides are responsible for a directed deposition of lignin on the cell walls, and this lignin in turn is concurrently an efficient strengthening material and protects the plants against pathogens. PMID- 21639842 TI - Hierarchical classification of glycoside hydrolases. AB - This review deals with structural and functional features of glycoside hydrolases, a widespread group of enzymes present in almost all living organisms. Their catalytic domains are grouped into 120 amino acid sequence-based families in the international classification of the carbohydrate-active enzymes (CAZy database). At a higher hierarchical level some of these families are combined in 14 clans. Enzymes of the same clan have common evolutionary origin of their genes and share the most important functional characteristics such as composition of the active center, anomeric configuration of cleaved glycosidic bonds, and molecular mechanism of the catalyzed reaction (either inverting, or retaining). There are now extensive data in the literature concerning the relationship between glycoside hydrolase families belonging to different clans and/or included in none of them, as well as information on phylogenetic protein relationship within particular families. Summarizing these data allows us to propose a multilevel hierarchical classification of glycoside hydrolases and their homologs. It is shown that almost the whole variety of the enzyme catalytic domains can be brought into six main folds, large groups of proteins having the same three-dimensional structure and the supposed common evolutionary origin. PMID- 21639843 TI - Role of lipid components in formation and reactivation of Mycobacterium smegmatis "nonculturable" cells. AB - We have found that transition of actively dividing Mycobacterium smegmatis cells into the dormant "nonculturable" state is accompanied by increase in the protein/lipid ratio and disappearance of one of the main lipid components of the mycobacterial cells, trehalose monomycolate. In this case, oleic acid is accumulated in the culture medium due to its secretion by the mycobacterial cells. Addition of lipids of different classes to "nonculturable" M. smegmatis cells induces their resuscitation. The lipid reactivating effect is evidently caused by the presence of fatty acids in their composition, because free fatty acids also exhibited reactivation effect. Oleic acid in concentration of 0.05-3 MUg/ml exhibited maximal effect, and that allows us to draw a conclusion concerning its signal role in the transition of dormant cells into active state. PMID- 21639844 TI - Rice dehydrin K-segments have in vitro antibacterial activity. AB - Dehydrins are groups of plant proteins that have been shown to response to various environmental stimuli such as dehydration, elevated salinity, and low temperature. However, their roles in plant defense against microbes have not been demonstrated. In an attempt to discover plant antimicrobial proteins, we have screened a rice cDNA library and isolated several cDNAs coding for dehydrins. Protein extracts from Escherichia coli expressing these cDNAs were tested for their activity against Gram-positive bacteria (Bacillus pumilus, B. subtilis, Staphylococcus aureus, and Sarcina lutea) and Gram-negative bacteria (Escherichia coli and Xanthomonas oryzae pv. oryzae). The results indicate that the crude protein extracts exhibited antibacterial activities against the Gram-positive bacteria. However, dehydrins purified by immunoaffinity chromatography were not active against the bacteria. To pinpoint the dehydrin peptides that were responsible for the bactericidal activity, we expressed DNA sequences coding for truncated dehydrins containing either K- or S-segment and found that K-segment peptides, and not S-segment, were responsible for the antibacterial activities against Gram-positive bacteria. Antibacterial assay with synthetic K-segments indicated that the peptides inhibited growth of B. pumilus with minimum inhibition concentration and minimum bactericidal concentration of 130 and 400 MUg/ml, respectively. PMID- 21639845 TI - CNG site-specific and methyl-sensitive endonuclease WEN1 from wheat seedlings. AB - Endonuclease WEN1 with apparent molecular mass about 27 kDa isolated from cytoplasmic vesicular fraction of aging coleoptiles of wheat seedlings has expressed site specificity action. This is a first detection and isolation of a site-specific endonuclease from higher eukaryotes, in general, and higher plants, in particular. The enzyme hydrolyzes deoxyribooligonucleotides of different composition on CNG (N is G, A, C, or T) sites by splitting the phosphodiester bond between C and N nucleotide residues in CNG sequence independent from neighbor nucleotide context except for CCCG. WEN1 prefers to hydrolyze methylated lambda phage DNA and double-stranded deoxyribooligonucleotides containing 5 methylcytosine sites (m(5)CAG, m(5)CTG) compared with unmethylated substrates. The enzyme is also able to hydrolyze single-stranded substrates, but in this case it splits unmethylated substrates predominantly. Detection in wheat seedlings of WEN1 endonuclease that is site specific, sensitive to the substrate methylation status, and modulated with S-adenosyl-L-methionine indicates that in higher plants restriction--modification systems or some of their elements, at least, may exist. PMID- 21639846 TI - Characterization of amino acid residues essential for tetramer formation and DNA binding activity of ssDNA-binding protein of Mycobacterium tuberculosis. AB - Mycobacterium tuberculosis Rv0054 encodes a single-stranded DNA-binding protein (MtbSSB) that is essential for survival of the human pathogen and causative agent of tuberculosis. The function of MtbSSB has been proposed to be different from its E. coli homolog. However, the critical amino acid residues of MtbSSB and their regulatory effects on DNA-binding ability remain to be clearly characterized. In this study, using a frequency-controlled random mutagenesis method (FRM), mutant libraries of MtbSSB were successfully constructed. On the whole, 146 single, double, and triple MtbSSB mutants, which covered 89% of the amino acid residues along the whole MtbSSB gene, were isolated. Using bacterial two-hybrid assays in combination with native PAGE assays, four new mutants, E62G, D104N, E94G/T137N, and S130P/G153N were found to totally or partially lose their ability to form tetramer. Three novel mutants, E62G, D104N, and E94G/T134N, were characterized to have a much lower ssDNA-binding activity, while one mutant, F21L, was found to have a significantly higher activity through both electrophoretic mobility shift and surface plasmon resonance assays. Interestingly, three amino acid residues, E62, D104, and E94, were found to regulate both oligomerization and ssDNA-binding activity of MtbSSB. Our work provides an important resource and should help improve the understanding of the biochemical mechanisms and structure-function relationship of the DNA-binding protein in this important human pathogen. PMID- 21639847 TI - Expression, purification, and characterization of putative Candida albicans Rad3, the product of orf19.7119. AB - Invasive infections of Candida albicans are life-threatening clinical conditions affecting immunosuppressed patients. To maintain genome integrity and diversity, C. albicans utilizes DNA repair systems, such as nucleotide excision repair (NER), to escape from attack by macrophages. Rad3 helicase is a component of the TFIIH complex, which plays a role in transcription and the NER pathway. Accumulated evidence of studies from Archaea to humans has revealed that the conserved structure, including an iron-containing domain, is essential in the function of Rad3 helicase activity. However, no study of the Rad3 protein of C. albicans has yet been reported. In the present study, putative C. albicans Rad3 (CaRad3) has been cloned with orf19.7119 of the Candida genome. CaRad3 proteins were over-expressed and purified from E. coli and S. cerevisiae using a Ni-NTA column and a size exclusion column for physicochemical and functional characterization. Through EMR and spectrometric analysis, we have proven that the purified CaRad3 protein has a Fe-S cluster. We also revealed that CaRad3 protein has a helicase activity on a duplex DNA substrate. Furthermore, we showed that the CaRad3 protein purified from yeasts was N-glycosylated, and that this protein complemented the defects in both the NER pathway and transcription. These data suggest that the Rad3 helicase in C. albicans is the product of the orf19.7119 gene. PMID- 21639848 TI - Free radical reaction products and antioxidant capacity in beating heart coronary artery surgery compared to conventional bypass. AB - Oxygen-derived free radicals are important agents of tissue injury during ischemia and reperfusion. The aim of this study was to investigate changes in protein and lipid oxidation and antioxidant status in beating heart coronary artery surgery and conventional bypass and to compare oxidative stress parameters between the two bypass methods. Serum lipid hydroperoxide, nitric oxide, protein carbonyl, nitrotyrosine, vitamin E, and beta-carotene levels and total antioxidant capacity were measured in blood of 30 patients undergoing beating heart coronary artery surgery (OPCAB, off-pump coronary artery bypass grafting) and 12 patients undergoing conventional bypass (CABG, on-pump coronary artery bypass grafting). In the OPCAB group, nitric oxide and nitrotyrosine levels decreased after reperfusion. Similarly, beta-carotene level and total antioxidant capacity also decreased after anesthesia and reperfusion. In the CABG group, nitric oxide and nitrotyrosine levels decreased after ischemia and reperfusion. However, protein carbonyl levels elevated after ischemia and reperfusion. Vitamin E, beta-carotene, and total antioxidant capacity decreased after ischemia and reperfusion. Significantly decreased nitration and impaired antioxidant status were seen after reperfusion in both groups. Moreover, elevated protein carbonyls were found in the CABG group. The off-pump procedure is associated with lower degree of oxidative stress than on-pump coronary surgery. PMID- 21639849 TI - Stability of haloalkaliphilic Geomicrobium sp. protease modulated by salt. AB - A novel protease from the halophilic bacterium Geomicrobium sp. EMB2 (MTCC 10310) is described. The activity of the protease was modulated by salt, and it exhibited remarkable stability in organic solvents, at alkaline pH, and in other denaturing conditions. The structural changes under various denaturing conditions were analyzed by measurements of intrinsic fluorescence and circular dichroism spectroscopy. Circular dichroism showed that the secondary structure of the protease was predominantly alpha-helical but unfolded in salt-free medium. The structure is regained by inclusion of NaCl in the range of 2-5%. The presence of NaCl exerted a protective effect against thermal, organic solvent, and guanidine hydrochloride denaturation by preventing unfolding. PMID- 21639850 TI - Purification, primary structure, and properties of Euphorbia characias latex purple acid phosphatase. AB - A purple acid phosphatase was purified to homogeneity from Euphorbia characias latex. The native protein has a molecular mass of 130 +/- 10 kDa and is formed by two apparently identical subunits, each containing one Fe(III) and one Zn(II) ion. The two subunits are connected by a disulfide bridge. The enzyme has an absorbance maximum at 540 nm, conferring a characteristic purple color due to a charge-transfer transition caused by a tyrosine residue (Tyr172) coordinated to the ferric ion. The cDNA nucleotide sequence contains an open reading frame of 1392 bp, and the deduced sequence of 463 amino acids shares a very high degree of identity (92-99%) to other purple acid phosphatases isolated from several higher plants. The enzyme hydrolyzes well p-nitrophenyl phosphate, a typical artificial substrate, and a broad range of natural phosphorylated substrates, such as ATP, ADP, glucose-6-phosphate, and phosphoenolpyruvate. The enzyme displays a pH optimum of 5.75 and is inhibited by molybdate, vanadate, and Zn2+, which are typical acid phosphatase inhibitors. PMID- 21639851 TI - Adaptive evolution signals in mitochondrial genes of Europeans. AB - Since modern Europeans appear to be descendants of the Late Pleistocene European peoples who survived the last glacial period, it is quite reasonable to expect the presence of adaptive genetic variants that originated in the Ice Age in the modern gene pool of Europeans. To find such adaptive variants, mitochondrial genomes have been analyzed of the modern population from Eastern and Central Europe belonging to haplogroups U4, U5, and V, that diversified during the Late Pleistocene and Holocene periods. Analysis of distribution of nonsynonymous and synonymous substitutions, as well as results of search for radical amino acid changes that arose under the influence of adaptation (positive destabilizing selection) allowed us to detect signals of molecular adaptation in different mitochondrial genes and haplogroups of mtDNA. However, there were very few strong adaptive signals (z > 3.09, P < 0.001) that could be due to the loss of adaptive mtDNA haplotypes during the Holocene warming. PMID- 21639852 TI - Structure of the O-polysaccharide of Providencia alcalifaciens O25 containing an amide of D-galacturonic acid with N(epsilon)-[(R)-1-carboxyethyl]-L-lysine. AB - An acidic O-polysaccharide was isolated by mild acid degradation of the lipopolysaccharide of Providencia alcalifaciens O25 followed by gel-permeation and anion-exchange chromatography. The O-polysaccharide was studied by sugar and methylation analyses along with (1)H and (13)C NMR spectroscopy, including two dimensional correlation (1)H,(13)C HMBC, and (1)H,(1)H ROESY experiments both in D(2)O and, to detect correlations for NH protons, in a 9 : 1 H(2)O/D(2)O mixture. An amino acid was isolated from the polysaccharide by acid hydrolysis and identified as N(epsilon)-[(R)-1-carboxyethyl]-L-lysine ("alaninolysine", 2S,8R alaLys) by determination of the specific optical rotation and (13)C NMR spectroscopy, using the authentic synthetic diastereomers 2S,8R-alaLys and 2S,8S alaLys for comparison. The structure of the branched tetrasaccharide repeating unit of the O-polysaccharide was established. PMID- 21639853 TI - Recombinant polyamine-binding protein of Synechocystis sp. PCC 6803 specifically binds to and is induced by polyamines. AB - His-tagged Synechocystis sp. PCC 6803 PotD protein (rPotD) involved in polyamine transport was overexpressed in Escherichia coli. The purified rPotD showed saturable binding kinetics with radioactively labeled polyamines. The rPotD exhibited a similar binding characteristic for three polyamines, with putrescine having less preference. The K(d) values for putrescine, spermine, and spermidine were 13.2, 8.3, and 7.8 uM, respectively. Binding of rPotD with polyamines was maximal at pH 8.0. Docking of these polyamines into the homology model of Synechocystis PotD showed that all three polyamines are able to interact with Synechocystis PotD. The binding modes of the docked putrescine and spermidine in Synechocystis are similar to those of PotF and PotD in E. coli, respectively. Competition experiments showed specific binding of rPotD with polyamines. The presence of putrescine and spermidine in the growth medium could induce an increase in PotD contents, suggesting the role of PotD in mediating the transport of polyamine in Synechocystis sp. PCC 6803. PMID- 21639854 TI - New approaches to identification and activity estimation of glyphosate degradation enzymes. AB - We propose a new set of approaches, which allow identifying the primary enzymes of glyphosate (N-phosphonomethyl-glycine) attack, measuring their activities, and quantitative analysis of glyphosate degradation in vivo and in vitro. Using the developed approach we show that glyphosate degradation can follow different pathways depending on physiological characteristics of metabolizing strains: in Ochrobactrum anthropi GPK3 the initial cleavage reaction is catalyzed by glyphosate-oxidoreductase with the formation of aminomethylphosphonic acid and glyoxylate, whereas Achromobacter sp. MPS12 utilize C-P lyase, forming sarcosine. The proposed methodology has several advantages as compared to others described in the literature. PMID- 21639855 TI - Mass and relative elution time profiling: two-dimensional analysis of sphingolipids in Alzheimer's disease brains. AB - Current lipidomic profiling methods rely mainly on MS to identify unknown lipids within a complex sample. We describe a new approach, involving LC*MS/MS (liquid chromatography*tandem MS) analysis of sphingolipids based on both mass and hydrophobicity, and use this method to characterize the SM (sphingomyelin), ceramide and GalCer (galactosylceramide) content of hippocampus from AD (Alzheimer's disease) and control subjects. Using a mathematical relationship we exclude the influence of sphingolipid mass on retention time, and generate two dimensional plots that facilitate accurate visualization and characterization of the different ceramide moieties within a given sphingolipid class, because related molecules align horizontally or vertically on the plots. Major brain GalCer species that differ in mass by only 0.04 Da were easily differentiated on the basis of their hydrophobicity. The importance of our method's capacity to define all of the major GalCer species in the brain samples is illustrated by the novel observation that the proportion of GalCer with hydroxylated fatty acids increased approximately 2-fold in the hippocampus of AD patients, compared with age- and gender-matched controls. This suggests activation of fatty acid hydroxylase in AD. Our method greatly improves the clarity of data obtained in a lipid profiling experiment and can be expanded to other lipid classes. PMID- 21639856 TI - Exo70, a subunit of the exocyst complex, interacts with SNEV(hPrp19/hPso4) and is involved in pre-mRNA splicing. AB - The Cdc5L (cell division cycle 5-like) complex is a spliceosomal subcomplex that also plays a role in DNA repair. The complex contains the splicing factor hPrp19, also known as SNEV or hPso4, which is involved in cellular life-span regulation and proteasomal breakdown. In a recent large-scale proteomics analysis for proteins associated with this complex, proteins involved in transcription, cell cycle regulation, DNA repair, the ubiquitin-proteasome system, chromatin remodelling, cellular aging, the cytoskeleton and trafficking, including four members of the exocyst complex, were identified. In the present paper we report that Exo70 interacts directly with SNEV(hPrp19/hPso4) and shuttles to the nucleus, where it associates with the spliceosome. We mapped the interaction site to the N-terminal 100 amino acids of Exo70, which interfere with pre-mRNA splicing in vitro. Furthermore, Exo70 influences the splicing of a model substrate as well as of its own pre-mRNA in vivo. In addition, we found that Exo70 is alternatively spliced in a cell-type- and cell-age- dependent way. These results suggest a novel and unexpected role of Exo70 in nuclear mRNA splicing, where it might signal membrane events to the splicing apparatus. PMID- 21639857 TI - Effect of dexamethasone on human osteoblasts in culture: involvement of beta1 integrin and integrin-linked kinase. AB - Adhesive interactions play a critical role in cell biology, influencing vital processes from proliferation to cell death. Integrins regulate cell-ECM (extracellular matrix) adhesion and must associate with phosphorylating proteins such as ILK (integrin-linked kinase). Dysregulation of ILK expression is associated with anchorage-independent growth, cell survival and inhibition of apoptosis. Glucocorticoids influence differentiation and adhesion of osteoblasts and can affect bone protein synthesis. The objective of this study was to analyse the effect of DEX (dexamethasone) on the biology of osteoblasts, together with its influence on the expression of ILK and beta1 integrin. For this, primary cultures of human osteoblasts were exposed to DEX at 10-9 M (physiological dose) and 10-6 M (pharmacological dose) for 24 and 48 h. Cell viability, apoptosis and cell adhesion were analysed, as well as protein expression of beta1 integrin and ILK. It was observed that cell viability and adhesion were reduced in the cultures evaluated. In comparison with the control cultures, there was slightly less apoptosis in the cultures exposed to the physiological dose and considerably more apoptosis in those exposed to the pharmacological dose. In all treated cultures, protein expression of ILK was slightly higher than in the control cultures, whereas that of beta1 integrin was significantly lower. Both proteins under study were co-localized at the cell periphery in all cultures. Our results suggest that DEX causes osteoblast anoikis, probably due to decreased beta1 integrin expression, which might have had a direct influence upon ILK, reducing its activation and preventing it from playing its characteristic anti-apoptotic role. PMID- 21639858 TI - Structural changes in the BH3 domain of SOUL protein upon interaction with the anti-apoptotic protein Bcl-xL. AB - The SOUL protein is known to induce apoptosis by provoking the mitochondrial permeability transition, and a sequence homologous with the BH3 (Bcl-2 homology 3) domains has recently been identified in the protein, thus making it a potential new member of the BH3-only protein family. In the present study, we provide NMR, SPR (surface plasmon resonance) and crystallographic evidence that a peptide spanning residues 147-172 in SOUL interacts with the anti-apoptotic protein Bcl-xL. We have crystallized SOUL alone and the complex of its BH3 domain peptide with Bcl-xL, and solved their three-dimensional structures. The SOUL monomer is a single domain organized as a distorted beta-barrel with eight anti parallel strands and two alpha-helices. The BH3 domain extends across 15 residues at the end of the second helix and eight amino acids in the chain following it. There are important structural differences in the BH3 domain in the intact SOUL molecule and the same sequence bound to Bcl-xL. PMID- 21639860 TI - Rural hospital ownership: medical service provision, market mix, and spillover effects. AB - OBJECTIVE: To test whether nonprofit, for-profit, or government hospital ownership affects medical service provision in rural hospital markets, either directly or through the spillover effects of ownership mix. DATA SOURCES/STUDY SETTING: Data are from the American Hospital Association, U.S. Census, CMS Healthcare Cost Report Information System and Prospective Payment System Minimum Data File, and primary data collection for geographic coordinates. The sample includes all nonfederal, general medical, and surgical hospitals located outside of metropolitan statistical areas and within the continental United States from 1988 to 2005. STUDY DESIGN: We estimate multivariate regression models to examine the effects of (1) hospital ownership and (2) hospital ownership mix within rural hospital markets on profitable versus unprofitable medical service offerings. PRINCIPAL FINDINGS: Rural nonprofit hospitals are more likely than for-profit hospitals to offer unprofitable services, many of which are underprovided services. Nonprofits respond less than for-profits to changes in service profitability. Nonprofits with more for-profit competitors offer more profitable services and fewer unprofitable services than those with fewer for-profit competitors. CONCLUSIONS: Rural hospital ownership affects medical service provision at the hospital and market levels. Nonprofit hospital regulation should reflect both the direct and spillover effects of ownership. PMID- 21639859 TI - Crystal structure of the glutamate receptor GluA1 N-terminal domain. AB - The AMPA (alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid) subfamily of iGluRs (ionotropic glutamate receptors) is essential for fast excitatory neurotransmission in the central nervous system. The malfunction of AMPARs (AMPA receptors) has been implicated in many neurological diseases, including Alzheimer's disease, Parkinson's disease and amyotrophic lateral sclerosis. The active channels of AMPARs and other iGluR subfamilies are tetramers formed exclusively by assembly of subunits within the same subfamily. It has been proposed that the assembly process is controlled mainly by the extracellular ATD (N-terminal domain) of iGluR. In addition, ATD has also been implicated in synaptogenesis, iGluR trafficking and trans-synaptic signalling, through unknown mechanisms. We report in the present study a 2.5 A (1 A=0.1 nm) resolution crystal structure of the ATD of GluA1. Comparative analyses of the structure of GluA1-ATD and other subunits sheds light on our understanding of how ATD drives subfamily-specific assembly of AMPARs. In addition, analysis of the crystal lattice of GluA1-ATD suggests a novel mechanism by which the ATD might participate in inter-tetramer AMPAR clustering, as well as in trans-synaptic protein-protein interactions. PMID- 21639861 TI - Effects of DTNBP1 genotype on brain development in children. AB - BACKGROUND: Schizophrenia is a neurodevelopmental disorder, and risk genes are thought to act through disruption of brain development. Several genetic studies have identified dystrobrevin-binding protein 1 (DTNBP1, also known as dysbindin) as a potential susceptibility gene for schizophrenia, but its impact on brain development is poorly understood. The present investigation examined for the first time the effects of DTNBP1 on brain structure in children. Our hypothesis was that a genetic variation in DTNBP1 (i.e., the single nucleotide polymorphism rs2619538) would be associated with differences in both gray and white matter brain regions previously implicated in schizophrenia. METHODS: Magnetic resonance imaging and voxel-based morphometry were used to examine brain structure in 52 male children aged between 10 and 12 years. Statistical inferences on the effects of DTNBP1 genotype on gray and white matter volume (GMV and WMV) were made at p < .05 after family-wise error correction for multiple comparisons across the whole brain. RESULTS: Individuals homozygous for the schizophrenia high-risk allele (AA) compared with those homozygous for the low-risk allele (TT) expressed reduced GMV in the left anterior cingulate gyrus and reduced WMV in the left medial frontal area. CONCLUSIONS: Our results suggest that genetic variation in DTNBP1 is associated with differences in gray and white matter; and that these effects are already evident in children as young as 10-12 years. These findings are consistent with the notion that the DTNBP1 genotype influences brain development and may thereby modulate vulnerability to schizophrenia. PMID- 21639862 TI - Characterization of the carbohydrate backbone of Vibrio parahaemolyticus O6 lipopolysaccharides. AB - Structural characterization studies have been carried out on the carbohydrate backbone of Vibrio parahaemolyticus serotype O6 lipopolysaccharides (LPS). The carbohydrate backbone isolated from O6 LPS by sequential derivatization, that is, dephosphorylation, O-deacylation, pyridylamination, N-deacylation and N acetylation, is a nonasaccharide consisting of 3 mol of D-glucosamine (GlcN) (of which one is pyridylaminated), 2 mol of L-glycero-D-manno-heptose (Hep), and 1 mol each of D-galactose (Gal), D-glucose (Glc), D-glucuronic acid (GlcA) and 3 deoxy-D-manno-oct-2-ulosonic acid (Kdo). Structural analyses by nuclear magnetic resonance spectroscopy and fast-atom bombardment mass spectrometry demonstrated that the carbohydrate backbone is beta-Galp-(1->2)-alpha-Hepp-(1->3)-alpha-Hepp (1->5)-alpha-Kdop-(2->6)-beta-GlcpNAc-(1->6)-GlcNAc-PA, in which the 3 substituted alpha-Hepp is further substituted by beta-GlcpNAc-(1->4)-beta-Glcp at position 4 and by beta-GlcpA at position 2. In native O6 LPS, an additional 1 mol of D-galacturonic acid, which is liberated by dephosphorylation in hydrofluoric acid, is present at an unknown position. A previous study by the present authors reported that, of 13 O-serotype LPS of V. parahaemolyticus, the only LPS from which Kdo was detected was from O6 LPS after mild acid hydrolysis. In the present study, we have demonstrated that only 1 mol of Kdo is present at the lipid A proximal position, a component which is common to the LPS in all serotypes of the bacterium, and that there is no additional Kdo in the carbohydrate backbone of O6 LPS. ELISA and ELISA inhibition analysis using antisera against O6 and Salmonella enterica Minnesota R595 and LPS of both strains further revealed that Kdo is not involved as an antigenic determinant of O6 LPS. PMID- 21639863 TI - Postpartum depression and help-seeking behaviors in immigrant Hispanic women. AB - OBJECTIVES: To describe perceptions of immigrant Hispanic women experiencing symptoms of postpartum depression (PPD) and to identify barriers to seeking mental health services. DESIGN: Qualitative descriptive. SETTING: Community health clinic. PARTICIPANTS: Twenty immigrant Hispanic women scoring positive for symptoms of PPD receiving health care at a community health clinic who declined mental health services participated in audiotaped interviews held in their homes. METHODS: Following Institutional Review Board approval and informed consent, interviews were conducted with study participants. Transcribed data were analyzed as appropriate for qualitative inquiry. RESULTS: Some of the women did not recognize and/or denied their symptoms attributing their sadness to financial concerns, family relationships, and/or work stressors. Study participants articulately described their symptoms and identified personal barriers including beliefs about emotional health, the perceived stigma of mental illness, hesitancy to seek treatment for symptoms of PPD, and cultural beliefs about motherhood and the role of women. Social barriers included inadequate social support, immigration status, and limited English proficiency. Health care delivery barriers included financial and time constraints and lack of child care and transportation. CONCLUSION: Limited social networks and barriers to health care should be addressed to foster positive outcomes. Mental health services should be embedded with primary health care or obstetric care clinics to facilitate access. Personal and professional support can make a significant contribution to the reduction of symptoms of PPD. PMID- 21639864 TI - Predicting the insertion length for gastric tube placement in neonates. AB - OBJECTIVE: To compare error rates of three existing methods of predicting the gastric tube insertion length in a group of neonates <1 month corrected age: age related, height-based (ARHB); direct distance nose-ear-xiphoid (NEX); and direct distance nose-ear-mid-umbilicus (NEMU). DESIGN: Randomized controlled trial. SETTING: Five neonatal care units in a large midwestern city. PARTICIPANTS: One hundred and seventy-three hospitalized neonates. METHODS: Neonates were randomly assigned to one of three groups: ARHB, NEX, or NEMU. For primary analysis, only tubes placed too high with the tube tip in the esophagus or at the gastroesophageal junction were considered to be misplaced. For secondary analysis, a stricter definition was used, and low placements (pylorus or duodenum) were also considered to be misplaced. All radiographs were blinded and read by a pediatric radiologist. RESULTS: For the primary analysis, the differences in percentages of correctly placed tubes among the three methods was statistically significant (chi(2) =34.45; p<.0001), with NEMU and ARHB more accurate than NEX (NEMU chi(2) =18.59, p<.0001; ARHB chi(2) =21.34, p<.0001). Using the stricter definition for placement, ARHB was not significantly different from NEX (p=.0615). A new ARHB equation was developed specific for neonates <1 month corrected age. CONCLUSIONS: Direct distance nose-ear-xiphoid should no longer be used as an nasogastric/orogastric (NG/OG) tube insertion-length predictor in neonates. Either NEMU for NG/OG tubes or the new ARHB equation for NG tubes should be used. PMID- 21639865 TI - Stress preconditioning of boar spermatozoa: a new approach to enhance semen quality. AB - Semen preparation and cryopreservation require finely adjusted procedures. Gametes are sensitive to environmental stresses, so in vitro procedures aim to minimize the inevitable harmful conditions. Applying stress to precondition cells has only been investigated recently. Studies demonstrated that by utilizing a well defined and properly applied hydrostatic pressure (HP) stress treatment to spermatozoa before in vitro storage, cryopreservation or insemination, cell survival and fertility improved compared with untreated controls. The birth of healthy piglets from treated fresh or frozen-thawed semen demonstrates the in vivo safety of the procedure. Although the biological mechanism is still unclear, several processes incorporating cellular stress response might explain the observations. This paper summarizes results, background, aspects and considerations of HP treatment for porcine semen. The new principle, i.e. to improve the stress tolerance by a defined sublethal stress may outline a new strategy in assisted reproductive technologies with unique theoretical and practical consequences. PMID- 21639866 TI - Neonatal lactic acidosis with methylmalonic aciduria due to novel mutations in the SUCLG1 gene. AB - BACKGROUND: Succinyl-coenzyme A ligase (SUCL) is a mitochondrial enzyme that catalyses the reversible conversion of succinyl-coenzyme A to succinate. SUCL consists of an alpha subunit, encoded by SUCLG1, and a beta subunit, encoded by either SUCLA2 or SUCLG2. Recently, mutations in SUCLG1 or SUCLA2 have been identified in patients with infantile lactic acidosis showing elevated urinary excretion of methylmalonate, mitochondrial respiratory chain (MRC) deficiency, and mitochondrial DNA depletion. METHODS: Case description of a Japanese female patient who manifested a neonatal-onset lactic acidosis with urinary excretion of methylmalonic acid. Enzymatic analyses (MRC enzyme assay and Western blotting) and direct sequencing analysis of SUCLA2 and SUCLG1 were performed. RESULTS: MRC enzyme assay and Western blotting showed that MRC complex I was deficient. SUCLG1 mutation analysis showed that the patient was a compound heterozygote for disease causing mutations (p.M14T and p.S200F). CONCLUSION: For patients showing neonatal lactic acidosis and prolonged mild methylmalonic aciduria, MRC activities and mutations of SUCLG1 or SUCLA2 should be screened for. PMID- 21639867 TI - Incidence of cardiovascular events after kidney transplantation and cardiovascular risk scores: study protocol. AB - BACKGROUND: Cardiovascular disease (CVD) is the major cause of death after renal transplantation. Not only conventional CVD risk factors, but also transplant specific risk factors can influence the development of CVD in kidney transplant recipients. The main objective of this study will be to determine the incidence of post-transplant CVD after renal transplantation and related factors. A secondary objective will be to examine the ability of standard cardiovascular risk scores (Framingham, REGICOR, SCORE, and DORICA) to predict post transplantation cardiovascular events in renal transplant recipients, and to develop a new score for predicting the risk of CVD after kidney transplantation. METHODS/DESIGN: Observational prospective cohort study of all kidney transplant recipients in the A Coruna Hospital (Spain) in the period 1981-2008 (2059 transplants corresponding to 1794 patients). The variables included will be: donor and recipient characteristics, chronic kidney disease-related risk factors, pre-transplant and post-transplant cardiovascular risk factors, routine biochemistry, and immunosuppressive, antihypertensive and lipid-lowering treatment. The events studied in the follow-up will be: patient and graft survival, acute rejection episodes and cardiovascular events (myocardial infarction, invasive coronary artery therapy, cerebral vascular events, new-onset angina, congestive heart failure, rhythm disturbances and peripheral vascular disease). Four cardiovascular risk scores were calculated at the time of transplantation: the Framingham score, the European Systematic Coronary Risk Evaluation (SCORE) equation, and the REGICOR (Registre Gironi del COR (Gerona Heart Registry)), and DORICA (Dyslipidemia, Obesity, and Cardiovascular Risk) functions. The cumulative incidence of cardiovascular events will be analyzed by competing risk survival methods. The clinical relevance of different variables will be calculated using the ARR (Absolute Risk Reduction), RRR (Relative Risk Reduction) and NNT (Number Needed to Treat). The ability of different cardiovascular risk scores to predict cardiovascular events will be analyzed by using the c index and the area under ROC curves. Based on the competing risks analysis, a nomogram to predict the probability of cardiovascular events after kidney transplantation will be developed. DISCUSSION: This study will make it possible to determine the post-transplant incidence of cardiovascular events in a large cohort of renal transplant recipients in Spain, to confirm the relationship between traditional and transplant-specific cardiovascular risk factors and CVD, and to develop a score to predict the risk of CVD in these patients. PMID- 21639868 TI - alpha-Mangostin extracted from the pericarp of the mangosteen (Garcinia mangostana Linn) reduces tumor growth and lymph node metastasis in an immunocompetent xenograft model of metastatic mammary cancer carrying a p53 mutation. AB - BACKGROUND: The mangosteen fruit has a long history of medicinal use in Chinese and Ayurvedic medicine. Recently, the compound alpha-mangostin, which is isolated from the pericarp of the fruit, was shown to induce cell death in various types of cancer cells in in vitro studies. This led us to investigate the antitumor growth and antimetastatic activities of alpha-mangostin in an immunocompetent xenograft model of mouse metastatic mammary cancer having a p53 mutation that induces a metastatic spectrum similar to that seen in human breast cancers. METHODS: Mammary tumors, induced by inoculation of BALB/c mice syngeneic with metastatic BJMC3879luc2 cells, were subsequently treated with alpha-mangostin at 0, 10 and 20 mg/kg/day using mini-osmotic pumps and histopathologically examined. To investigate the mechanisms of antitumor ability by alpha-mangostin, in vitro studies were also conducted. RESULTS: Not only were in vivo survival rates significantly higher in the 20 mg/kg/day alpha-mangostin group versus controls, but both tumor volume and the multiplicity of lymph node metastases were significantly suppressed. Apoptotic levels were significantly increased in the mammary tumors of mice receiving 20 mg/kg/day and were associated with increased expression of active caspase-3 and -9. Other significant effects noted at this dose level were decreased microvessel density and lower numbers of dilated lymphatic vessels containing intraluminal tumor cells in mammary carcinoma tissues. In vitro, alpha-mangostin induced mitochondria-mediated apoptosis and G1 phase arrest and S-phase suppression in the cell cycle. Since activation by Akt phosphorylation plays a central role in a variety of oncogenic processes, including cell proliferation, anti-apoptotic cell death, angiogenesis and metastasis, we also investigated alterations in Akt phosphorylation induced by alpha-mangostin treatment both in vitro and in vivo. Quantitative analysis and immunohistochemistry showed that alpha-mangostin significantly decreased the levels of phospho-Akt-threonine 308 (Thr308), but not serine 473 (Ser473), in both mammary carcinoma cell cultures and mammary carcinoma tissues in vivo. CONCLUSIONS: Since lymph node involvement is the most important prognostic factor in breast cancer patients, the antimetastatic activity of alpha-mangostin as detected in mammary cancers carrying a p53 mutation in the present study may have specific clinical applications. In addition, alpha-mangostin may have chemopreventive benefits and/or prove useful as an adjuvant therapy, or as a complementary alternative medicine in the treatment of breast cancer. PMID- 21639869 TI - Central nervous system mast cells in peripheral inflammatory nociception. AB - BACKGROUND: Functional aspects of mast cell-neuronal interactions remain poorly understood. Mast cell activation and degranulation can result in the release of powerful pro-inflammatory mediators such as histamine and cytokines. Cerebral dural mast cells have been proposed to modulate meningeal nociceptor activity and be involved in migraine pathophysiology. Little is known about the functional role of spinal cord dural mast cells. In this study, we examine their potential involvement in nociception and synaptic plasticity in superficial spinal dorsal horn. Changes of lower spinal cord dura mast cells and their contribution to hyperalgesia are examined in animal models of peripheral neurogenic and non neurogenic inflammation. RESULTS: Spinal application of supernatant from activated cultured mast cells induces significant mechanical hyperalgesia and long-term potentiation (LTP) at spinal synapses of C-fibers. Lumbar, thoracic and thalamic preparations are then examined for mast cell number and degranulation status after intraplantar capsaicin and carrageenan. Intradermal capsaicin induces a significant percent increase of lumbar dural mast cells at 3 hours post administration. Peripheral carrageenan in female rats significantly increases mast cell density in the lumbar dura, but not in thoracic dura or thalamus. Intrathecal administration of the mast cell stabilizer sodium cromoglycate or the spleen tyrosine kinase (Syk) inhibitor BAY-613606 reduce the increased percent degranulation and degranulated cell density of lumbar dural mast cells after capsaicin and carrageenan respectively, without affecting hyperalgesia. CONCLUSION: The results suggest that lumbar dural mast cells may be sufficient but are not necessary for capsaicin or carrageenan-induced hyperalgesia. PMID- 21639870 TI - Elevated IL-6 receptor expression on CD4+ T cells contributes to the increased Th17 responses in patients with chronic hepatitis B. AB - BACKGROUND: Increased numbers of Interleukin-17-producing CD4+ T cells (Th17) have been found in association with hepatitis B virus (HBV)-induced liver injury. However, the mechanism underlying the increase of Th17 responses in patients with HBV infection remains unclear. In this study, we investigate the possible regulatory mechanisms of increased Th17 responses in patients with chronic hepatitis B(CHB). METHODS: Th17 response and IL-6R expression on CD4+ T cells in peripheral blood samples were determined by flow cytometry. Cytokines TGF-beta, IL-1beta, IL-6 and IL-17 in plasma and/or supernatant samples were determined by ELISA and the IL-17 and IL-6R mRNA levels were quantified by quantitative real time reverse polymerase chain reaction. RESULTS: All these data indicated that the frequency of periphery Th17 cells is significantly correlated with the percentage of CD4+ T cells expressing IL-6R in CHB patients. CD4+ T cells from patients with CHB, but not those from healthy donors, produced higher levels of IL-17 and had more IL-6R expression upon stimulation with the HBV core antigen (HBcAg) in vitro. The PMA/ionomycin and HBcAg -stimulated up-regulation of IL-17 production by CD4+ T cells could be reversed by a neutralizing antibody against IL-6R. CONCLUSION: we showed that enhancement of IL-6R expression on CD4+ T cells upon HBV infection contributes to increased Th17 response in patients with CHB. PMID- 21639871 TI - Systematic development of a communication skills training course for physicians performing work disability assessments: from evidence to practice. AB - BACKGROUND: Physicians require specific communication skills, because the face-to face contact with their patients is an important source of information. Although physicians who perform work disability assessments attend some communication related training courses during their professional education, no specialised and evidence-based communication skills training course is available for them. Therefore, the objectives of this study were: 1) to systematically develop a training course aimed at improving the communication skills of physicians during work disability assessment interviews with disability claimants, and 2) to plan an evaluation of the training course. METHODS: A physician-tailored communication skills training course was developed, according to the six steps of the Intervention Mapping protocol. Data were collected from questionnaire studies among physicians and claimants, a focus group study among physicians, a systematic review of the literature, and meetings with various experts. Determinants and performance objectives were formulated. A concept version of the training course was discussed with several experts before the final training course programme was established. The evaluation plan was developed by consulting experts, social insurance physicians, researchers, and policy-makers, and discussing with them the options for evaluation. RESULTS: A two-day post-graduate communication skills training course was developed, aimed at improving professional communication during work disability assessment interviews. Special focus was on active teaching strategies, such as practising the skills in role play. An adoption and implementation plan was formulated, in which the infrastructure of the educational department of the institute that employs the physicians was utilised. Improvement in the skills and knowledge of the physicians who will participate in the training course will be evaluated in a randomised controlled trial. CONCLUSIONS: The feasibility and practical relevance of the communication skills training course that was developed seem promising. Such a course may be relevant for physicians in many countries who perform work disability assessments. The development of the first training course of this type represents an important advancement in this field. PMID- 21639872 TI - Safety, tolerability, and impact on allergic inflammation of autologous E.coli autovaccine in the treatment of house dust mite asthma--a prospective open clinical trial. AB - BACKGROUND: Asthma is increasing worldwide and results from a complex immunological interaction between genetic susceptibility and environmental factors. Autovaccination with E. coli induces a strong TH-1 immune response, thus offering an option for the treatment of allergic diseases. METHODS: Prospective open trial on safety, tolerability, and impact on allergic inflammation of an autologous E.coli autovaccine in intermittent or mild persistent house dust mite asthma. Determination of exhaled nitric monoxide (eNO) before and after bronchial mite challenge initially and after nine months of autovaccination. RESULTS: In nine subjects and a total of 306 injections, we observed 101 episodes of local erythema (33.3%; median of maximal diameter 2.5 cm), 95 episodes of local swelling (31.1%; median of maximal diameter 3 cm), and 27 episodes of local pain (8.8%). Four subjects reported itching at the injection site with a total of 30 episodes (9.8%). Median eNO increase after autovaccination was significantly smaller (from 27.3 to 33.8 ppb; p = 0.334) compared to initial values (from 32.6 to 42.2 ppb; p = 0.046) (p = 0.034). We observed no serious adverse events. All organ functions (inclusive electrocardiogramm) and laboratory testing of the blood (clinical chemistry, hematology) and the urine (screening test, Beta microglobuline) were within normal limits. Vital signs undulated within the physiological variability. CONCLUSION: The administration of autologous autovacine for the treatment of house dust mite asthma resulted in a reduction of the eNO increase upon bronchial mite challenge. In nine subjects and 306 injections, only a few mild local reactions and no systemic severe adverse events were observed. TRIAL REGISTRATION: EudraCT Nr. 2005-005534-12ClinicalTrials.gov ID NCT00677209. PMID- 21639873 TI - Uptake and outcomes of a prevention-of mother-to-child transmission (PMTCT) program in Zomba district, Malawi. AB - BACKGROUND: HIV prevalence among pregnant women in Malawi is 12.6%, and mother-to child transmission is a major route of transmission. As PMTCT services have expanded in Malawi in recent years, we sought to determine uptake of services, HIV-relevant infant feeding practices and mother-child health outcomes. METHODS: A matched-cohort study of HIV-infected and HIV-uninfected mothers and their infants at 18-20 months post-partum in Zomba District, Malawi. 360 HIV-infected and 360 HIV-uninfected mothers were identified through registers. 387 mother child pairs were included in the study. RESULTS: 10% of HIV-infected mothers were on HAART before delivery, 27% by 18-20 months post-partum. sd-NVP was taken by 75% of HIV-infected mothers not on HAART, and given to 66% of infants. 18% of HIV infected mothers followed all current recommended PMTCT options. HIV-infected mothers breastfed fewer months than HIV-uninfected mothers (12 vs.18, respectively; p < 0.01). 19% of exposed versus 5% of unexposed children had died by 18-20 months; p < 0.01. 28% of exposed children had been tested for HIV prior to the study, 76% were tested as part of the study and 11% were found HIV positive. HIV-free survival by 18-20 months was 66% (95%CI 58-74). There were 11(6%) maternal deaths among HIV-infected mothers only. CONCLUSION: This study shows low PMTCT program efficiency and effectiveness under routine program conditions in Malawi. HIV-free infant survival may have been influenced by key factors, including underuse of HAART, underuse of sd-NVP, and suboptimal infant feeding practices. Maternal mortality among HIV-infected women demands attention; improved maternal survival is a means to improve infant survival. PMID- 21639874 TI - Interference with work in fibromyalgia: effect of treatment with pregabalin and relation to pain response. AB - BACKGROUND: Clinical trials in chronic pain often collect information about interference with work as answers to component questions of commonly used questionnaires but these data are not normally analysed separately. METHODS: We performed a meta-analysis of individual patient data from four large trials of pregabalin for fibromyalgia lasting 8-14 weeks. We analysed data on interference with work, inferred from answers to component questions of Fibromyalgia Impact Questionnaire (FIQ), Short Form 36 Health Survey, Sheehan Disability Scale, and Multidimensional Assessment of Fatigue, including "How many days in the past week did you miss work, including housework, because of fibromyalgia?" from FIQ. Analyses were performed according to randomised treatment group (pregabalin 150 600 mg daily or placebo), pain improvement (0-10 numerical pain rating scale scores at trial beginning vs. end), and end of trial pain state (100 mm visual analogue pain scale [VAS]). RESULTS: Comparing treatment group average outcomes revealed modest improvement over the duration of the trials, more so with active treatment than with placebo. For the 'work missed' question from FIQ the change for patients on placebo was from 2.2 (standard deviation [SD] 2.3) days of work lost per week at trial beginning to 1.9 (SD 2.1) days lost at trial end (p < 0.01). For patients on 600 mg pregabalin the change was from 2.1 (SD 2.2) days to 1.6 (SD 2.0) days (p < 0.001). However, the change in days of work lost was substantial in patients with a good pain response: from 2.0 (SD 2.2) days to 0.97 (SD 1.6) days (p < 0.0001) for those experiencing >/= 50% pain improvement and from 1.9 (SD 2.2) days to 0.73 (SD 1.4) days (p < 0.0001) for those achieving a low level of pain at trial end (<30 mm on the VAS). Patients achieving both >/= 50% pain improvement and a pain score <30 mm on the VAS had the largest improvement, from 2.0 (SD 2.2) days to 0.60 (SD 1.3) days (p < 0.0001). Analysing answers to the other questions yielded qualitatively similar results. CONCLUSIONS: Effective pain treatment goes along with benefit regarding work. A reduction in time off work >1 day per week can be achieved in patients with good pain responses. PMID- 21639875 TI - Elevated pre-treatment levels of plasma C-reactive protein are associated with poor prognosis after breast cancer: a cohort study. AB - INTRODUCTION: We examined whether plasma C-reactive protein (CRP) levels at the time of diagnosis of breast cancer are associated with overall survival, disease free survival, death from breast cancer, and recurrence of breast cancer. METHODS: We observed 2,910 women for up to seven years after they were diagnosed with invasive breast cancer (median follow-up time was three years). Plasma levels of high-sensitivity CRP were measured at the time of diagnosis and we assessed the association between CRP levels and risk of reduced overall and disease-free survival, death from breast cancer, and recurrence of breast cancer by using the Kaplan-Meier method and Cox proportional hazards regression. During follow-up, 383 women died (225 of whom died from breast cancer) and 118 women experienced recurrence of breast cancer. RESULTS: Elevated CRP levels across tertiles at the time of diagnosis were associated with reduced overall and disease-free survival and with increased risk of death from breast cancer (log rank trend for all, P < 0.001), but not with recurrence. The multifactor-adjusted hazard ratios (HR) of reduced overall survival among women in the middle and highest versus the lowest tertile of CRP were 1.30 (95% CI, 0.97 to 1.73) and 1.94 (1.48 to 2.55), respectively. Corresponding HRs of reduced disease-free survival were 1.16 (0.89 to 1.50) and 1.76 (1.38 to 2.25) and of death from breast cancer 1.22 (0.84 to 1.78) and 1.66 (1.15 to 2.41). Dividing CRP levels into octiles resulted in a stepwise increased risk of reduced overall survival (P for trend <0.001) and the multifactor-adjusted HR among women in the highest versus the lowest octile of CRP was 2.51 (1.53 to 4.12). Compared to women with CRP levels in the 0 to 25% percentile (<0.78 mg/L), the multifactor-adjusted HR of reduced overall survival among women with CRP levels >=95% percentile (>=16.4 mg/L) was 3.58 (2.36 to 5.42). Among women with HER2-positive tumours, the multifactor-adjusted HR of reduced overall survival for the highest versus the lowest tertile of CRP was 8.63 (2.04 to 36.4). CONCLUSIONS: Elevated CRP levels at the time of diagnosis of breast cancer are associated with reduced overall and disease-free survival and with increased risk of death from breast cancer. PMID- 21639876 TI - Protocol for translabial 3D-ultrasonography for diagnosing levator defects (TRUDIL): a multicentre cohort study for estimating the diagnostic accuracy of translabial 3D-ultrasonography of the pelvic floor as compared to MR imaging. AB - BACKGROUND: Pelvic organ prolapse (POP) is a condition affecting more than half of the women above age 40. The estimated lifetime risk of needing surgical management for POP is 11%. In patients undergoing POP surgery of the anterior vaginal wall, the re-operation rate is 30%. The recurrence risk is especially high in women with a levator ani defect. Such defect is present if there is a partially or completely detachment of the levator ani from the inferior ramus of the symphysis. Detecting levator ani defects is relevant for counseling, and probably also for treatment. Levator ani defects can be imaged with MRI and also with Translabial 3D ultrasonography of the pelvic floor. The primary aim of this study is to assess the diagnostic accuracy of translabial 3D ultrasonography for diagnosing levator defects in women with POP with Magnetic Resonance Imaging as the reference standard. Secondary goals of this study include quantification of the inter-observer agreement about levator ani defects and determining the association between levator defects and recurrent POP after anterior repair. In addition, the cost-effectiveness of adding translabial ultrasonography to the diagnostic work-up in patients with POP will be estimated in a decision analytic model. METHODS/DESIGN: A multicentre cohort study will be performed in nine Dutch hospitals. 140 consecutive women with a POPQ stage 2 or more anterior vaginal wall prolapse, who are indicated for anterior colporapphy will be included. Patients undergoing additional prolapse procedures will also be included. Prior to surgery, patients will undergo MR imaging and translabial 3D ultrasound examination of the pelvic floor. Patients will be asked to complete validated disease specific quality of life questionnaires before surgery and at six and twelve months after surgery. Pelvic examination will be performed at the same time points. Assuming a sensitivity and specificity of 90% of 3D ultrasound for diagnosing levator defects in a population of 120 women with POP, with a prior probability of levator ani defects of 40%, we will be able to estimate predictive values with good accuracy (i.e. confidence limits of at most 10% below or above the point estimates of positive and negative predictive values).Anticipating 3% unclassifiable diagnostic images because of technical reasons, and a further safety margin of 10% we plan to recruit 140 patients. TRIAL REGISTRATION: Nederlands trial register NTR2220. PMID- 21639877 TI - No Paragonimus in high-risk groups in Cote d'Ivoire, but considerable prevalence of helminths and intestinal protozoon infections. AB - BACKGROUND: Paragonimiasis is a neglected tropical disease caused by an infection with lung flukes that is transmitted through the consumption of undercooked crabs. The disease is often confused with tuberculosis. Paragonimiasis is thought to be endemic in south-western Cote d'Ivoire. METHODS: Two cross-sectional surveys were carried out in the first half of 2009 in patients attending two tuberculosis centres of Abidjan. A third cross-sectional survey was conducted in May 2010 in children of two primary schools in Dabou, where crabs are frequently consumed. Patients with chronic cough provided three sputum samples plus one stool sample. Sputum samples were examined for tuberculosis with an auramine staining technique and for Paragonimus eggs using a concentration technique. Stool samples were subjected to the Ritchie technique. Schoolchildren provided a single stool sample, and samples were subjected to the Kato-Katz and an ether concentration technique. A pre-tested questionnaire was administered to patients and schoolchildren to investigate food consumption habits. Additionally, between June 2009 and August 2010, shellfish were purchased from markets in Abidjan and Dabou and examined for metacercariae. RESULTS: No human case of paragonimiasis was diagnosed. However, trematode infections were seen in 32 of the 272 shellfish examined (11.8%). Questionnaire results revealed that crab and pig meat is well cooked before consumption. Among the 278 patients with complete data records, 62 had tuberculosis, with a higher prevalence in males than females (28.8% vs. 13.9%, chi2 = 8.79, p = 0.003). The prevalence of helminths and intestinal protozoa was 4.6% and 16.9%, respectively. In the school survey, among 166 children with complete data records, the prevalence of helminths and intestinal protozoa was 22.3% and 48.8%, respectively. Boys had significantly higher prevalences of helminths and intestinal protozoa than girls. Hookworm was the predominant helminth species and Entamoeba coli was the most common intestinal protozoon species (13.8%). CONCLUSIONS: Not a single case of Paragonimus was found in two high-risk groups of Cote d'Ivoire, most likely explained by food consumption habits. However, other helminth and intestinal protozoon infections were common. PMID- 21639878 TI - The correlates of leisure time physical activity among an adults population from southern Taiwan. AB - BACKGROUND: Assessing the correlates of practicing physical activity during leisure time is important with regard to planning and designing public health strategies to increase beneficial behaviors among adult populations. Although the importance of leisure time physical activity (LTPA) is highlighted in many Western countries, there are not many publications on physical activity patterns, and even less on their correlates, in non-Western societies. The goal of this study was thus to explore the determinants influencing adults' leisure time physical activity (LTPA) in a city in southern Taiwan. METHODS: A cross-sectional population-based study was conducted in 2007, using a standardized questionnaire. Energy expenditure was dichotomized into two groups based on the recommended levels of moderate physical activity from LTPA: >=10 or < 10 MET.hr.wk(-1). Logistic regression analyses were applied to the results. RESULTS: A total of 762 subjects with valid data took part in the study (mean age 53.8 +/- 13.8 years). In multivariate logistic regression analysis, we found the following results: Age was positively associated with LTPA. Adults with stronger perceived convenience of exercise facilities (OR = 2.04; 95%CI = 1.28-3.24) and past exercise experience in school (OR = 1.86; 95%CI= 1.19-2.91) participated in more LTPA. Subjects with more general social support (OR = 1.66;95%CI = 1.13-2.44), greater knowledge about the health benefits of exercise (OR = 1.85;95%CI = 1.25-2.74), more sports media consumption (OR = 1.94;95%CI = 1.26-2.98), and higher self efficacy (OR = 3.99;95%CI = 2.67-5.97) were more likely to engage in LTPA. Further analysis comparing different sources of social support showed only social support from friends had a significant positive association (OR = 1.73;95%CI = 1.14-2.63) with increased LTPA. CONCLUSIONS: LTPA in southern city of Taiwan showed some unique associations with age, socioeconomic status and media consumption that are not commonly reported in the Western World and similar associations with regards to psychosocial correlates of LTPA participation. Further studies from developing countries are warranted to highlight culture specific differences in physical activity participation. PMID- 21639879 TI - Non-leisure time physical activity is an independent predictor of longevity for a Taiwanese elderly population: an eight-year follow-up study. AB - BACKGROUND: The aim of this study is to determine the relationship between leisure time physical activity (LTPA) and non-leisure time physical activity (NLTPA) on mortality among the elderly in Taiwan. METHODS: This is a prospective observational cohort study. We analyzed the mortality data from a cohort of 876 non-institutionalized community-dwelling men and women aged 65 years or over, who were recruited by stratified clustering random sampling from Tainan city and participated in the 1996 Elderly Medication Survey. Information about activities and other variables were collected by structured interviews at baseline in the participants' home. The Cox proportional hazards model and crude death rate were applied to estimate mortality risk. RESULTS: Among the 876 participants, 312 died during the follow-up period (1996-2004). In the unadjusted Cox regression model, subjects aged over 75, having difficulty in carrying out activities of daily living (ADLs), a BMI less than 18.5, a history of diabetes mellitus or stroke, without LTPA or being inactive in NLTPA, were found to have a higher risk of eight-year mortality. With the adjustment for age, gender, education level, habitual smoking and drinking, living status, BMI and medical history, the mortality was found to be higher among the sedentary subjects, either defined by lack of LTPA or NLTPA, with the hazard ratio of 1.27 (95% confidence interval [CI] = 0.97-1.66) and 1.45 (95% CI = 1.07-1.97), respectively. Furthermore, when both LTPA and NLTPA were put into the model simultaneously, NLTPA (HR = 1.40; 95% CI = 1.03-1.91) but not LTPA (HR = 1.21, 95% CI = 0.92-1.59) significantly predicted mortality during eight-year follow-up. In addition, subjects who were actively engaged in NLTPA had a lower mortality risk especially in subjects without performing LTPA. CONCLUSIONS: NLTPA is an independent predictor of longevity among older people in Taiwan. A physically active lifestyle, especially engaged in NLTPA, is associated with lower mortality risk in the elderly population. We thus suggest that encouraging older people to keep on engaging in customary NLTPA is good for their health. PMID- 21639880 TI - Increased expression of cystine/glutamate antiporter in multiple sclerosis. AB - BACKGROUND: Glutamate excitotoxicity contributes to oligodendrocyte and tissue damage in multiple sclerosis (MS). Intriguingly, glutamate level in plasma and cerebrospinal fluid of MS patients is elevated, a feature which may be related to the pathophysiology of this disease. In addition to glutamate transporters, levels of extracellular glutamate are controlled by cystine/glutamate antiporter x(c)-, an exchanger that provides intracellular cystine for production of glutathione, the major cellular antioxidant. The objective of this study was to analyze the role of the system x(c)- in glutamate homeostasis alterations in MS pathology. METHODS: Primary cultures of human monocytes and the cell line U-937 were used to investigate the mechanism of glutamate release. Expression of cystine glutamate exchanger (xCT) was quantified by quantitative PCR, Western blot, flow cytometry and immunohistochemistry in monocytes in vitro, in animals with experimental autoimmune encephalomyelitis (EAE), the animal model of MS, and in samples of MS patients. RESULTS AND DISCUSSION: We show here that human activated monocytes release glutamate through cystine/glutamate antiporter x(c)- and that the expression of the catalytic subunit xCT is upregulated as a consequence of monocyte activation. In addition, xCT expression is also increased in EAE and in the disease proper. In the later, high expression of xCT occurs both in the central nervous system (CNS) and in peripheral blood cells. In particular, cells from monocyte-macrophage-microglia lineage have higher xCT expression in MS and in EAE, indicating that immune activation upregulates xCT levels, which may result in higher glutamate release and contribution to excitotoxic damage to oligodendrocytes. CONCLUSIONS: Together, these results reveal that increased expression of the cystine/glutamate antiporter system x(c)- in MS provides a link between inflammation and excitotoxicity in demyelinating diseases. PMID- 21639881 TI - FXTAS is rare among Portuguese patients with movement disorders: FMR1 premutations may be associated with a wider spectrum of phenotypes. AB - The fragile X-associated tremor/ataxia syndrome (FXTAS) is a late-onset neurodegenerative disorder caused by expansions of 55-200 CGG repeats in the 5'UTR of the FMR1 gene. These FMR1 premutation expansions have relatively high frequency in the general population. To estimate the frequency of FMR1 premutations among Portuguese males with non-familial, late-onset movement disorders of unknown etiology, we assessed CGG repeat size in males with disease onset after the age of 50 and negative or unknown family history for late-onset movement disorders, who were sent for SCA, HD, or PD genetic testing at a reference laboratory. The selected patients had a primary clinical diagnosis based on one of the following cardinal features of FXTAS: ataxia, tremor, or cognitive decline. A total of 86 subjects were genotyped for the CGG repeat in the FMR1 gene. We detected one patient with an expansion in the premutation range. The frequency of FMR1 premutations was 1.9% (1/54) in our group of patients with ataxia as the primary clinical feature, and 1.2% (1/86) in the larger movement disorders group. In the family of the FXTAS case, premutation transmitting females presented a history of psychiatric symptoms, suggesting that, given the wide phenotypical expression of the premutation in females, neuropsychiatric surveillance is necessary. In conclusion, genetic testing for FXTAS should be made available to patients with adult-onset movement disorders to enable adequate genetic counseling to family members. PMID- 21639882 TI - A sub-cubic time algorithm for computing the quartet distance between two general trees. AB - BACKGROUND: When inferring phylogenetic trees different algorithms may give different trees. To study such effects a measure for the distance between two trees is useful. Quartet distance is one such measure, and is the number of quartet topologies that differ between two trees. RESULTS: We have derived a new algorithm for computing the quartet distance between a pair of general trees, i.e. trees where inner nodes can have any degree >= 3. The time and space complexity of our algorithm is sub-cubic in the number of leaves and does not depend on the degree of the inner nodes. This makes it the fastest algorithm so far for computing the quartet distance between general trees independent of the degree of the inner nodes. CONCLUSIONS: We have implemented our algorithm and two of the best competitors. Our new algorithm is significantly faster than the competition and seems to run in close to quadratic time in practice. PMID- 21639883 TI - Did changing primary care delivery models change performance? A population based study using health administrative data. AB - BACKGROUND: Primary care reform in Ontario, Canada started with the introduction of new enrollment models, the two largest of which are Family Health Networks (FHNs), a capitation-based model, and Family Health Groups (FHGs), a blended fee for-service model. The purpose of this study was to evaluate differences in performance between FHNs and FHGs and to compare performance before and after physicians joined these new primary care groups. METHODS: This study used Ontario administrative claims data to compare performance measures in FHGs and FHNs. The study population included physicians who belonged to a FHN or FHG for at least two years. Patients were included in the analyses if they enrolled with a physician in the two years after the physician joined a FHN or FHG, and also if they saw the physician in a two year period prior to the physician joining a FHN or FHG. Performance was derived from the administrative data, and included measures of preventive screening for cancer (breast, cervical, colorectal) and chronic disease management (diabetes, heart failure, asthma). RESULTS: Performance measures did not vary consistently between models. In some cases, performance approached current benchmarks (Pap smears, mammograms). In other cases it was improving in relation to previous measures (colorectal cancer screening). There were no changes in screening for cervical cancer or breast cancer after joining either a FHN or FHG. Colorectal cancer screening increased in both FHNs and FHGs. After enrolling in either a FHG or a FHN, prescribing performance measures for diabetes care improved. However, annual eye examinations decreased for younger people with diabetes after joining a FHG or FHN. There were no changes in performance measures for heart failure management or asthma care after enrolling in either a FHG or FHN. CONCLUSIONS: Some improvements in preventive screening and diabetes management which were seen amongst people after they enrolled may be attributed to incentive payments offered to physicians within FHGs and FHNs. However, these primary care delivery models need to be compared with other delivery models and fee for service practices in order to describe more specifically what aspects of model delivery and incentives affect care. PMID- 21639884 TI - Staying at work with chronic nonspecific musculoskeletal pain: a qualitative study of workers' experiences. AB - BACKGROUND: Many people with chronic nonspecific musculoskeletal pain (CMP) have decreased work ability. The majority, however, stays at work despite their pain. Knowledge about workers who stay at work despite chronic pain is limited, narrowing our views on work participation. The aim of this study was to explore why people with CMP stay at work despite pain (motivators) and how they manage to maintain working (success factors). METHODS: A semi-structured interview was conducted among 21 subjects who stay at work despite CMP. Participants were included through purposeful sampling. Interviews were audio-recorded, transcribed verbatim, and imported into computer software Atlas.ti. Data was analyzed by means of thematic analysis. The interviews consisted of open questions such as: "Why are you working with pain?" or "How do you manage working while having pain?" RESULTS: A total of 16 motivators and 52 success factors emerged in the interviews. Motivators were categorized into four themes: work as value, work as therapy, work as income generator, and work as responsibility. Success factors were categorized into five themes: personal characteristics, adjustment latitude, coping with pain, use of healthcare services, and pain beliefs. CONCLUSIONS: Personal characteristics, well-developed self-management skills, and motivation to work may be considered to be important success factors and prerequisites for staying at work, resulting in behaviors promoting staying at work such as: raising adjustment latitude, changing pain-coping strategies, organizing modifications and conditions at work, finding access to healthcare services, and asking for support. Motivators and success factors for staying at work may be used for interventions in rehabilitation and occupational medicine, to prevent absenteeism, or to promote a sustainable return to work. This qualitative study has evoked new hypotheses about staying at work; quantitative studies on staying at work are needed to obtain further evidence. PMID- 21639885 TI - Molecular evolution of rbcL in three gymnosperm families: identifying adaptive and coevolutionary patterns. AB - BACKGROUND: The chloroplast-localized ribulose-1, 5-biphosphate carboxylase/oxygenase (Rubisco), the primary enzyme responsible for autotrophy, is instrumental in the continual adaptation of plants to variations in the concentrations of CO2. The large subunit (LSU) of Rubisco is encoded by the chloroplast rbcL gene. Although adaptive processes have been previously identified at this gene, characterizing the relationships between the mutational dynamics at the protein level may yield clues on the biological meaning of such adaptive processes. The role of such coevolutionary dynamics in the continual fine-tuning of RbcL remains obscure. RESULTS: We used the timescale and phylogenetic analyses to investigate and search for processes of adaptive evolution in rbcL gene in three gymnosperm families, namely Podocarpaceae, Taxaceae and Cephalotaxaceae. To understand the relationships between regions identified as having evolved under adaptive evolution, we performed coevolutionary analyses using the software CAPS. Importantly, adaptive processes were identified at amino acid sites located on the contact regions among the Rubisco subunits and on the interface between Rubisco and its activase. Adaptive amino acid replacements at these regions may have optimized the holoenzyme activity. This hypothesis was pinpointed by evidence originated from our analysis of coevolution that supported the correlated evolution between Rubisco and its activase. Interestingly, the correlated adaptive processes between both these proteins have paralleled the geological variation history of the concentration of atmospheric CO2. CONCLUSIONS: The gene rbcL has experienced bursts of adaptations in response to the changing concentration of CO2 in the atmosphere. These adaptations have emerged as a result of a continuous dynamic of mutations, many of which may have involved innovation of functional Rubisco features. Analysis of the protein structure and the functional implications of such mutations put forward the conclusion that this evolutionary scenario has been possible through a complex interplay between adaptive mutations, often structurally destabilizing, and compensatory mutations. Our results unearth patterns of evolution that have likely optimized the Rubisco activity and uncover mutational dynamics useful in the molecular engineering of enzymatic activities. REVIEWERS: This article was reviewed by Prof. Christian Blouin (nominated by Dr W Ford Doolittle), Dr Endre Barta (nominated by Dr Sandor Pongor), and Dr Nicolas Galtier. PMID- 21639886 TI - Effect of 50% and maximal inspired oxygen concentrations on respiratory variables in isoflurane-anesthetized horses. AB - BACKGROUND: The purpose of this study was to compare the effects of 0.5 fraction of inspired oxygen (FiO2) and >0.95 FiO2 on pulmonary gas exchange, shunt fraction and oxygen delivery (DO2) in dorsally recumbent horses during inhalant anesthesia. The use of 0.5 FiO2 has the potential to reduce absorption atelectasis (compared to maximal FiO2) and augment alveolar oxygen (O2) tensions (compared to ambient air) thereby improving gas exchange and DO2. Our hypothesis was that 0.5 FiO2 would reduce ventilation-perfusion mismatching and increase the fraction of pulmonary blood flow that is oxygenated, thus improving arterial oxygen content and DO2. RESULTS: Arterial partial pressures of O2 were significantly higher than preanesthetic levels at all times during anesthesia in the >0.95 FiO2 group. Arterial partial pressures of O2 did not change from preanesthetic levels in the 0.5 FiO2 group but were significantly lower than in the >0.95 FiO2 group from 15 to 90 min of anesthesia. Alveolar to arterial O2 tension difference was increased significantly in both groups during anesthesia compared to preanesthetic values. The alveolar to arterial O2 tension difference was significantly higher at all times in the >0.95 FiO2 group compared to the 0.5 FiO2 group. Oxygen delivery did not change from preanesthetic values in either group during anesthesia but was significantly lower than preanesthetic values 10 min after anesthesia in the 0.5 FiO2 group. Shunt fraction increased in both groups during anesthesia attaining statistical significance at varying times. Shunt fraction was significantly increased in both groups 10 min after anesthesia but was not different between groups. Alveolar dead space ventilation increased after 3 hr of anesthesia in both groups. CONCLUSIONS: Reducing FiO2 did not change alveolar dead space ventilation or shunt fraction in dorsally recumbent, mechanically ventilated horses during 3 hr of isoflurane anesthesia. Reducing FiO2 in dorsally recumbent isoflurane anesthetized horses does not improve oxygenation or oxygen delivery. PMID- 21639887 TI - Validation of the Edinburgh Claudication Questionnaire in 1st generation Black African-Caribbean and South Asian UK migrants: a sub-study to the Ethnic Echocardiographic Heart of England Screening (E-ECHOES) study. AB - BACKGROUND: We determined the diagnostic accuracy of the Edinburgh Claudication Questionnaire (ECQ) in 1st generation Black African-Caribbean UK migrants as previous diagnostic questionnaires have been found to be less accurate in this population. We also determined the diagnostic accuracy of translated versions of the ECQ in 1st generation South Asian UK migrants, as this has not been investigated before. METHODS: Subjects were recruited from the Ethnic Echocardiographic Heart of England Screening (E-ECHOES) study, a community based screening survey for heart failure in minority ethnic groups. Translated versions of the ECQ were prepared following a recognised protocol. All participants attending screening between October 2007 and February 2009 were asked to complete the ECQ in the language of their choice (English, Punjabi, Bengali, Urdu, Hindi or Gujarati). Subjects answering positively to experiencing leg pain or discomfort on walking were asked to return to have Ankle Brachial Pressure Index (ABPI) measured. RESULTS: 154 out of 2831 subjects participating in E-ECHOES (5.4%) were eligible to participate in this sub-study, for which 74.3% returned for ABPI assessment. Non-responders were younger than participants (59[9] vs. 65[11] years; p=0.015). Punjabi, English and Bengali questionnaires identified participants with Intermittent Claudication, so these questionnaires were assessed. The sensitivities (SN), specificities (SP), positive (PPV) and negative (NPV) predictive values were calculated. English: SN: 50%; SP: 68%; PPV: 43%; NPV: 74%. Punjabi: SN: 50%; SP: 87%; PPV: 43%; NPV: 90%. Bengali: SN: 33%; SP: 50%; PPV: 13%; NPV: 73%. There were significant differences in diagnostic accuracy between the 3 versions (Punjabi: 83.8%; Bengali: 45%; English: 62.2%; p<0.0001). No significant differences were found in sensitivity and specificity between illiterate and literate participants in any of the questionnaires and there was no significant different difference between those under and over 60 years of age. CONCLUSIONS: Our findings suggest that the ECQ is not as sensitive or specific a diagnostic tool in 1st generation Black African-Caribbean and South Asian UK migrants than in the Edinburgh Artery Study, reflecting the findings of other diagnostic questionnaires in these minority ethnic groups. However this study is limited by sample size so conclusions should be interpreted with caution. PMID- 21639888 TI - Client preferences and acceptability for medical abortion and MVA as early pregnancy termination method in northwest Ethiopia. AB - BACKGROUND: Increasing access to safe abortion services is the most effective way of preventing the burden of unsafe abortion, which is achieved by increasing safe choices for pregnancy termination. Medical abortion for termination of early abortion is said to safe, effective, and acceptable to women in several countries. In Ethiopia, however, medical methods have, until recently, never been used. For this reason it is important to assess women's preferences and the acceptability of medical abortion and manual vacuum aspiration (MVA) in the early first trimester pregnancy termination and factors affecting acceptability of medical and MVA abortion services. METHODS: A prospective study was conducted in two hospitals and two clinics from March 2009 to November 2009. The study population consisted of 414 subjects over the age of 18 with intrauterine pregnancies of up to 63 days' estimated gestation. Of these 251 subjects received mifepristone and misoprostol and 159 subjects received MVA. Questionnaires regarding expectations and experiences were administered before the abortion and at the 2-week follow-up visit. RESULTS: The study groups were similar with respect to age, marital status, educational status, religion and ethnicity. Their mean age was about 23, majority in both group completed secondary education and about half were married. Place of residence and duration of pregnancy were associated with method choice. Subjects undergoing medical abortions reported significantly greater satisfaction than those undergoing surgical abortions (91.2% vs 82.4%; P < .001). Of those women who had medical abortion, (83.3%) would choose the method again if needed, and (77.4%) of those who had MVA would also choose the method again. Ninety four percent of women who had medical abortion and 86.8% of those who had MVA would recommend the method to their friends. CONCLUSIONS: Women receiving medical abortion were more satisfied with their method and more likely to choose the same method again than were subjects undergoing surgical abortion. We conclude that medical abortion can be used widely as an alternative method for early pregnancy termination. PMID- 21639889 TI - Analysis of complex metabolic behavior through pathway decomposition. AB - BACKGROUND: Understanding complex systems through decomposition into simple interacting components is a pervasive paradigm throughout modern science and engineering. For cellular metabolism, complexity can be reduced by decomposition into pathways with particular biochemical functions, and the concept of elementary flux modes provides a systematic way for organizing metabolic networks into such pathways. While decomposition using elementary flux modes has proven to be a powerful tool for understanding and manipulating cellular metabolism, its utility, however, is severely limited since the number of modes in a network increases exponentially with its size. RESULTS: Here, we present a new method for decomposition of metabolic flux distributions into elementary flux modes. Our method can easily operate on large, genome-scale networks since it does not require all relevant modes of the metabolic network to be generated. We illustrate the utility of our method for metabolic engineering of Escherichia coli and for understanding the survival of Mycobacterium tuberculosis (MTB) during infection. CONCLUSIONS: Our method can achieve computational time improvements exceeding 2000-fold and requires only several seconds to generate elementary mode decompositions on genome-scale networks. These improvements arise from not having to generate all relevant elementary modes prior to initiating the decomposition. The decompositions from our method are useful for understanding complex flux distributions and debugging genome-scale models. PMID- 21639890 TI - A novel class of heat-responsive small RNAs derived from the chloroplast genome of Chinese cabbage (Brassica rapa). AB - BACKGROUND: Non-coding small RNAs play critical roles in various cellular processes in a wide spectrum of eukaryotic organisms. Their responses to abiotic stress have become a popular topic of economic and scientific importance in biological research. Several studies in recent years have reported a small number of non-coding small RNAs that map to chloroplast genomes. However, it remains uncertain whether small RNAs are generated from chloroplast genome and how they respond to environmental stress, such as high temperature. Chinese cabbage is an important vegetable crop, and heat stress usually causes great losses in yields and quality. Under heat stress, the leaves become etiolated due to the disruption and disassembly of chloroplasts. In an attempt to determine the heat-responsive small RNAs in chloroplast genome of Chinese cabbage, we carried out deep sequencing, using heat-treated samples, and analysed the proportion of small RNAs that were matched to chloroplast genome. RESULTS: Deep sequencing provided evidence that a novel subset of small RNAs were derived from the chloroplast genome of Chinese cabbage. The chloroplast small RNAs (csRNAs) include those derived from mRNA, rRNA, tRNA and intergenic RNA. The rRNA-derived csRNAs were preferentially located at the 3'-ends of the rRNAs, while the tRNA-derived csRNAs were mainly located at 5'-termini of the tRNAs. After heat treatment, the abundance of csRNAs decreased in seedlings, except those of 24 nt in length. The novel heat-responsive csRNAs and their locations in the chloroplast were verified by Northern blotting. The regulation of some csRNAs to the putative target genes were identified by real-time PCR. Our results reveal that high temperature suppresses the production of some csRNAs, which have potential roles in transcriptional or post-transcriptional regulation. CONCLUSIONS: In addition to nucleus, the chloroplast is another important organelle that generates a number of small RNAs. Many members of csRNA families are highly sensitive to heat stress. Some csRNAs respond to heat stress by silencing target genes. We suggest that proper temperature is important for production of chloroplast small RNAs, which are associated with plant resistance to abiotic stress. PMID- 21639891 TI - Computer-based fluorescence quantification: a novel approach to study nucleolar biology. AB - BACKGROUND: Nucleoli are composed of possibly several thousand different proteins and represent the most conspicuous compartments in the nucleus; they play a crucial role in the proper execution of many cellular processes. As such, nucleoli carry out ribosome biogenesis and sequester or associate with key molecules that regulate cell cycle progression, tumorigenesis, apoptosis and the stress response. Nucleoli are dynamic compartments that are characterized by a constant flux of macromolecules. Given the complex and dynamic composition of the nucleolar proteome, it is challenging to link modifications in nucleolar composition to downstream effects. RESULTS: In this contribution, we present quantitative immunofluorescence methods that rely on computer-based image analysis. We demonstrate the effectiveness of these techniques by monitoring the dynamic association of proteins and RNA with nucleoli under different physiological conditions. Thus, the protocols described by us were employed to study stress-dependent changes in the nucleolar concentration of endogenous and GFP-tagged proteins. Furthermore, our methods were applied to measure de novo RNA synthesis that is associated with nucleoli. We show that the techniques described here can be easily combined with automated high throughput screening (HTS) platforms, making it possible to obtain large data sets and analyze many of the biological processes that are located in nucleoli. CONCLUSIONS: Our protocols set the stage to analyze in a quantitative fashion the kinetics of shuttling nucleolar proteins, both at the single cell level as well as for a large number of cells. Moreover, the procedures described here are compatible with high throughput image acquisition and analysis using HTS automated platforms, thereby providing the basis to quantify nucleolar components and activities for numerous samples and experimental conditions. Together with the growing amount of information obtained for the nucleolar proteome, improvements in quantitative microscopy as they are described here can be expected to produce new insights into the complex biological functions that are orchestrated by the nucleolus. PMID- 21639892 TI - Brachypodium distachyon: a new pathosystem to study Fusarium head blight and other Fusarium diseases of wheat. AB - BACKGROUND: Fusarium species cause Fusarium head blight (FHB) and other important diseases of cereals. The causal agents produce trichothecene mycotoxins such as deoxynivalenol (DON). The dicotyledonous model species Arabidopsis thaliana has been used to study Fusarium-host interactions but it is not ideal for model-to crop translation. Brachypodium distachyon (Bd) has been proposed as a new monocotyledonous model species for functional genomic studies in grass species. This study aims to assess the interaction between the most prevalent FHB-causing Fusarium species and Bd in order to develop and exploit Bd as a genetic model for FHB and other Fusarium diseases of wheat. RESULTS: The ability of Fusarium graminearum and Fusarium culmorum to infect a range of Bd tissues was examined in various bioassays which showed that both species can infect all Bd tissues examined, including intact foliar tissues. DON accumulated in infected spike tissues at levels similar to those of infected wheat spikes. Histological studies revealed details of infection, colonisation and host response and indicate that hair cells are important sites of infection. Susceptibility to Fusarium and DON was assessed in two Bd ecotypes and revealed variation in resistance between ecotypes. CONCLUSIONS: Bd exhibits characteristics of susceptibility highly similar to those of wheat, including susceptibility to spread of disease in the spikelets. Bd is the first reported plant species to allow successful infection on intact foliar tissues by FHB-causing Fusarium species. DON appears to function as a virulence factor in Bd as it does in wheat. Bd is proposed as a valuable model for undertaking studies of Fusarium head blight and other Fusarium diseases of wheat. PMID- 21639893 TI - Down regulation of E-Cadherin (ECAD) - a predictor for occult metastatic disease in sentinel node biopsy of early squamous cell carcinomas of the oral cavity and oropharynx. AB - BACKGROUND: Prognostic factors in predicting occult lymph node metastasis in patients with head and neck squamous-cell carcinoma (HNSCC) are necessary to improve the results of the sentinel lymph node procedure in this tumour type. The E-Cadherin glycoprotein is an intercellular adhesion molecule in epithelial cells, which plays an important role in establishing and maintaining intercellular connections. OBJECTIVES: To determine the value of the molecular marker E-Cadherin in predicting regional metastatic disease. METHODS: E-Cadherin expression in tumour tissue of 120 patients with HNSCC of the oral cavity and oropharynx were evaluated using the tissue microarray technique. 110 tumours were located in the oral cavity (91.7%; mostly tongue), 10 tumours in the oropharynx (8.3%). Intensity of E-Cadherin expression was quantified by the Intensity Reactivity Score (IRS). These results were correlated with the lymph node status of biopsied sentinel lymph nodes. Univariate and multivariate analysis was used to determine statistical significance. RESULTS: pT-stage, gender, tumour side and location did not correlate with lymph node metastasis. Differentiation grade (p = 0.018) and down regulation of E-Cadherin expression significantly correlate with positive lymph node status (p = 0.005) in univariate and multivariate analysis. CONCLUSION: These data suggest that loss of E-cadherin expression is associated with increased lymhogeneous metastasis of HNSCC. E-cadherin immunohistochemistry may be used as a predictor for lymph node metastasis in squamous cell carcinoma of the oral cavity and oropharynx. LEVEL OF EVIDENCE: 2b. PMID- 21639894 TI - "If the patients decide not to tell what can we do?"- TB/HIV counsellors' dilemma on partner notification for HIV. AB - BACKGROUND: There is a global consensus towards universal access to human immunodeficiency virus (HIV) services consequent to the increasing availability of antiretroviral therapy. However, to benefit from these services, knowledge of one's HIV status is critical. Partner notification for HIV is an important component of HIV counselling because it is an effective strategy to prevent secondary transmission, and promote early diagnosis and prompt treatment of HIV patients' sexual partners. However, counsellors are often frustrated by the reluctance of HIV-positive patients to voluntarily notify their sexual partners. This study aimed to explore tuberculosis (TB)/HIV counsellors' perspectives regarding confidentiality and partner notification. METHODS: Qualitative research interviews were conducted in the Northwest Region of Cameroon with 30 TB/HIV counsellors in 4 treatment centres, and 2 legal professionals between September and December 2009. Situational Analysis (positional map) was used for data analysis. RESULTS: Confidentiality issues were perceived to be handled properly despite concerns about patients' reluctance to report cases of violation due to apprehension of reprisals from health care staffs. All the respondents encouraged voluntary partner notification, and held four varying positions when confronted with patients who refused to voluntarily notify their partners. Position one focused on absolute respect of patients' autonomy; position two balanced between the respect of patients' autonomy and their partners' safety; position three wished for protection of sexual partners at risk of HIV infection and legal protection for counsellors; and position four requested making HIV testing and partner notification routine processes. CONCLUSION: Counsellors regularly encounter ethical, legal and moral dilemmas between respecting patients' confidentiality and autonomy, and protecting patients' sexual partners at risk of HIV infection.This reflects the complexity of partner notification and demonstrates that no single approach is optimal, but instead certain contextual factors and a combination of different approaches should be considered. Meanwhile, adopting a human rights perspective in HIV programmes will balance the interests of both patients and their partners, and ultimately enhance universal access to HIV services. PMID- 21639895 TI - Media awards for responsible reporting of suicide: Experiences from Australia, Belgium and Denmark. AB - BACKGROUND: Media awards to encourage responsible reporting of suicide have been introduced in several countries, including Australia, Belgium and Denmark. AIMS: This study aimed to examine the experiences of Australian, Belgian and Danish award recipients in preparing stories on suicide, and consider the impacts of the awards for these recipients and for media professionals more broadly. METHOD: We conducted semi-structured telephone interviews with the majority (14 out of 15) of past recipients of the awards in the three countries of interest. RESULTS: Media awards appear to show promise as a method of reinforcing national and international media guidelines on reporting suicide. The recipients of awards were proud to have had their achievements recognized in this way, and had developed a heightened awareness of the issues inherent in reporting suicide. Although relatively few had prepared subsequent stories on suicide, a number had been given opportunities to provide advice to other media professionals about how best to approach this sensitive topic. Recipients viewed the awards as an important means by which good quality reporting can be rewarded, and a springboard for raising community awareness about suicide. CONCLUSION: The experience from Australia, Belgium and Denmark suggests that media awards which recognize responsible reporting of suicide are extremely worthwhile. PMID- 21639896 TI - Chromite oxidation by manganese oxides in subseafloor basalts and the presence of putative fossilized microorganisms. AB - Chromite is a mineral with low solubility and is thus resistant to dissolution. The exception is when manganese oxides are available, since they are the only known naturally occurring oxidants for chromite. In the presence of Mn(IV) oxides, Cr(III) will oxidise to Cr(VI), which is more soluble than Cr(III), and thus easier to be removed. Here we report of chromite phenocrysts that are replaced by rhodochrosite (Mn(II) carbonate) in subseafloor basalts from the Koko Seamount, Pacific Ocean, that were drilled and collected during the Ocean Drilling Program (ODP) Leg 197. The mineral succession chromite-rhodochrosite saponite in the phenocrysts is interpreted as the result of chromite oxidation by manganese oxides. Putative fossilized microorganisms are abundant in the rhodochrosite and we suggest that the oxidation of chromite has been mediated by microbial activity. It has previously been shown in soils and in laboratory experiments that chromium oxidation is indirectly mediated by microbial formation of manganese oxides. Here we suggest a similar process in subseafloor basalts. PMID- 21639897 TI - Evaluation of the impact of interdisciplinarity in cancer care. AB - BACKGROUND: Teamwork is a key component of the health care renewal strategy emphasized in Quebec, elsewhere in Canada and in other countries to enhance the quality of oncology services. While this innovation would appear beneficial in theory, empirical evidences of its impact are limited. Current efforts in Quebec to encourage the development of local interdisciplinary teams in all hospitals offer a unique opportunity to assess the anticipated benefits. These teams working in hospital outpatient clinics are responsible for treatment, follow-up and patient support. The study objective is to assess the impact of interdisciplinarity on cancer patients and health professionals. METHODS/DESIGN: This is a quasi-experimental study with three comparison groups distinguished by intensity of interdisciplinarity: strong, moderate and weak. The study will use a random sample of 12 local teams in Quebec, stratified by intensity of interdisciplinarity. The instrument to measure the intensity of the interdisciplinarity, developed in collaboration with experts, encompasses five dimensions referring to aspects of team structure and process. Self-administered questionnaires will be used to measure the impact of interdisciplinarity on patients (health care utilization, continuity of care and cancer services responsiveness) and on professionals (professional well-being, assessment of teamwork and perception of teamwork climate). Approximately 100 health professionals working on the selected teams and 2000 patients will be recruited. Statistical analyses will include descriptive statistics and comparative analysis of the impact observed according to the strata of interdisciplinarity. Fixed and random multivariate statistical models (multilevel analyses) will also be used. DISCUSSION: This study will pinpoint to what extent interdisciplinarity is linked to quality of care and meets the complex and varied needs of cancer patients. It will ascertain to what extent interdisciplinary teamwork facilitated the work of professionals. Such findings are important given the growing prevalence of cancer and the importance of attracting and retaining health professionals to work with cancer patients. PMID- 21639898 TI - The CDK9/cyclin T1 subunits of P-TEFb in mouse oocytes and preimplantation embryos: a possible role in embryonic genome activation. AB - BACKGROUND: Two stages of genome activation have been identified in the mouse embryo. Specifically, minor transcriptional activation is evident at the one-cell stage and a second major episode of activation occurs at the two-cell stage. Nuclear translocation of RNA polymerase II and phosphorylation of the C-terminal domain (CTD) of the largest enzyme subunit are major determinants of embryonic genome activation. P-TEFb, the Pol II CTD kinase, regulates transcriptional elongation via phosphorylation of the serine 2 residues of the CTD. RESULTS: Here, we show that the CDK9 and cyclin T1 subunits of P-TEFb are present in mouse oocytes and preimplantation embryos. Both proteins translocate to pronuclei at the late one-cell stage and are predominantly localized in nuclei at the two-cell stage. We additionally examine the effects of the CDK9-specific inhibitor, flavopiridol, on mouse preimplantation development. Our data show that treatment with the drug results in mislocalization of CDK9, cyclin T1, and phosphorylated Pol II, as well as developmental arrest at the two-cell stage. CONCLUSIONS: A change in CDK9 localization from the cytoplasm to the pronucleus occurs at the time of minor embryonic genome activation, and CDK9 accumulation at the two-cell stage is evident, concomitant with major transcriptional activation of the embryonic genome. Moreover, CDK9 inhibition triggers a developmental block at the two-cell stage. Our findings clearly indicate that CDK9 is essential for embryonic genome activation in the mouse. PMID- 21639899 TI - Echocardiographic predictors of early in-hospital heart failure during first ST elevation acute myocardial infarction: does myocardial performance index and left atrial volume improve diagnosis over conventional parameters of left ventricular function? AB - BACKGROUND: Left ventricular ejection fraction (LVEF) has been considered a major determinant of early outcome in acute myocardial infarction (AMI). Myocardial performance index (MPI) has been associated to early evolution in AMI in a heterogeneous population, including non ST-elevation or previous AMI. Left atrial volume has been related with late evolution after AMI. We evaluated the independent role of clinical and echocardiographic variables including LVEF, MPI and left atrial volume in predicting early in-hospital congestive heart failure (CHF) specifically in patients with a first isolated ST-elevation AMI. METHODS: Echocardiography was performed within 30 hours of chest pain in 95 patients with a first ST-elevation AMI followed during the first week of hospitalization. Several clinical and echocardiographic variables were analyzed. CHF was defined as Killip class >= II. Multivariate regression analysis was used to select independent predictor of in-hospital CHF. RESULTS: Early in-hospital CHF occurred in 29 (31%) of patients. LVEF <= 0.45 was the single independent and highly significant predictor of early CHF among other clinical and echocardiographic variables (odds ratio 17.0; [95% CI 4.1 - 70.8]; p < 0.0001). MPI alone could not predict CHF in first ST-elevation AMI patients. Left atrial volume was not associated with early CHF in such patients. CONCLUSION: For patients with first, isolated ST-elevation AMI, LVEF assessed by echocardiography still constitutes a strong and accurate independent predictor of early in-hospital CHF, superior to isolated MPI and left atrial volume in this particular subset of patients. PMID- 21639900 TI - Design and methods for a randomized clinical trial comparing three outreach efforts to improve screening mammography adherence. AB - BACKGROUND: Despite the demonstrated need to increase screening mammography utilization and strong evidence that mail and telephone outreach to women can increase screening, most managed care organizations have not adopted comprehensive outreach programs. The uncertainty about optimum strategies and cost effectiveness have retarded widespread acceptance. While 70% of women report getting a mammogram within the prior 2 years, repeat mammography rates are less than 50%. This 5-year study is conducted though a Central Massachusetts healthcare plan and affiliated clinic. All women have adequate health insurance to cover the test. METHODS/DESIGN: This randomized study compares 3 arms: reminder letter alone; reminder letter plus reminder call; reminder letter plus a second reminder and booklet plus a counselor call. All calls provide women with the opportunity to schedule a mammogram in a reasonable time. The invention period will span 4 years and include repeat attempts. The counselor arm is designed to educate, motivate and counsel women in an effort to alleviate PCP burden.All women who have been in the healthcare plan for 24 months and who have a current primary care provider (PCP) and who are aged 51-84 are randomized to 1 of 3 arms. Interventions are limited to women who become >= 18 months from a prior mammogram. Women and their physicians may opt out of the intervention study.Measurement of completed mammograms will use plan billing records and clinic electronic records. The primary outcome is the proportion of women continuously enrolled for >= 24 months who have had >= 1 mammogram in the last 24 months. Secondary outcomes include the number of women who need repeat interventions. The cost effectiveness analysis will measure all costs from the provider perspective. DISCUSSION: So far, 18,509 women aged 51-84 have been enrolled into our tracking database and were randomized into one of three arms. At baseline, 5,223 women were eligible for an intervention. We anticipate that the outcome will provide firm data about the maximal effectiveness as well as the cost effectiveness of the interventions both for increasing the mammography rate and the repeat mammography rate. TRIAL REGISTRATION: http://clinicaltrials.gov/NCT01332032. PMID- 21639901 TI - Bilateral ovarian teratoma complicated with carcinosarcoma in a 68 year old woman: a case report. AB - BACKGROUND: Composing of less than 1% of all ovarian cancers, immature teratoma is a malignancy that mainly affects the young, and they present with advanced disease. The treatment of immature teratoma is conservative primary surgery usually involving unilateral salpingo-oophorectomy followed by combination chemotherapy. CASE PRESENTATION: Here we present a case of a 68 year old woman with bilateral ovarian teratoma complicated with carcinosarcoma. The patient was diagnosed as FIGO stage IIIC. She underwent neoadjuvant chemotherapy and interval cytoreduction followed by optimal cytoreduction. The post operative management strategies and gynaecological follow up studies revealed no evidence of regional or distant metastasis. CONCLUSION: Thus the choice of initial treatment should be decided in a selective fashion depending on various prognostic factors in order to increase the survival of the patients. PMID- 21639902 TI - Understanding competing risks: a simulation point of view. AB - BACKGROUND: Competing risks methodology allows for an event-specific analysis of the single components of composite time-to-event endpoints. A key feature of competing risks is that there are as many hazards as there are competing risks. This is not always well accounted for in the applied literature. METHODS: We advocate a simulation point of view for understanding competing risks. The hazards are envisaged as momentary event forces. They jointly determine the event time. Their relative magnitude determines the event type. 'Empirical simulations' using data from a recent study on cardiovascular events in diabetes patients illustrate subsequent interpretation. The method avoids concerns on identifiability and plausibility known from the latent failure time approach. RESULTS: The 'empirical simulations' served as a proof of concept. Additionally manipulating baseline hazards and treatment effects illustrated both scenarios that require greater care for interpretation and how the simulation point of view aids the interpretation. The simulation algorithm applied to real data also provides for a general tool for study planning. CONCLUSIONS: There are as many hazards as there are competing risks. All of them should be analysed. This includes estimation of baseline hazards. Study planning must equally account for these aspects. PMID- 21639903 TI - Thymic plasmacytoid dendritic cells are susceptible to productive HIV-1 infection and efficiently transfer R5 HIV-1 to thymocytes in vitro. AB - BACKGROUND: HIV-1 infection of the thymus contributes to the defective regeneration and loss of CD4+ T cells in HIV-1-infected individuals. As thymic dendritic cells (DC) are permissive to infection by HIV-1, we examined the ability of thymic DC to enhance infection of thymocytes which may contribute to the overall depletion of CD4+ T cells. We compared productive infection in isolated human thymic and blood CD11c+ myeloid DC (mDC) and CD123+ plasmacytoid DC (pDC) using enhanced green fluorescent protein (EGFP) CCR5 (R5)-tropic NL(AD8) and CXCR4 (X4)-tropic NL4-3 HIV-1 reporter viruses. Transfer of productive HIV-1 infection from thymic mDC and pDC was determined by culturing these DC subsets either alone or with sorted thymocytes. RESULTS: Productive infection was observed in both thymic pDC and mDC following exposure to R5 HIV-1 and X4 HIV-1. Thymic pDC were more frequently productively infected by both R5 and X4 HIV-1 than thymic mDC (p = 0.03; n = 6). Thymic pDC efficiently transferred productive R5 HIV-1 infection to both CD3(hi) (p = 0.01; mean fold increase of 6.5; n = 6) and CD3(lo) thymocytes (mean fold increase of 1.6; n = 2). In comparison, transfer of productive infection by thymic mDC was not observed for either X4 or R5 HIV-1. CONCLUSIONS: The capacity of thymic pDC to efficiently transfer R5 HIV 1 to both mature and immature thymocytes that are otherwise refractory to R5 virus may represent a pathway to early infection and impaired production of thymocytes and CD4+ T cells in HIV-1-infected individuals. PMID- 21639904 TI - Molecular characterization and phylogenetic analysis of small ruminant lentiviruses isolated from Canadian sheep and goats. AB - BACKGROUND: Small Ruminant Lentiviruses (SRLV) are widespread in Canadian sheep and goats and represent an important health issue in these animals. There is however no data about the genetic diversity of Caprine Arthritis Encephalitis Virus (CAEV) or Maedi Visna Virus (MVV) in this country. FINDINGS: We performed a molecular and phylogenetic analysis of sheep and goat lentiviruses from a small geographic area in Canada using long sequences from the gag region of 30 infected sheep and 36 infected goats originating from 14 different flocks. Pairwise DNA distance and phylogenetic analyses revealed that all SRLV sequences obtained from sheep clustered tightly with prototypical Maedi visna sequences from America. Similarly, all SRLV strains obtained from goats clustered tightly with prototypical US CAEV-Cork strain. CONCLUSIONS: The data reported in this study suggests that Canadian and US SRLV strains share common origins. In addition, the molecular data failed to bring to light any evidence of past cross species transmission between sheep and goats, which is consistent with the type of farming practiced in this part of the country where single species flocks predominate and where opportunities of cross species transmissions are proportionately low. PMID- 21639905 TI - Insulin alleviates degradation of skeletal muscle protein by inhibiting the ubiquitin-proteasome system in septic rats. AB - Hypercatabolism is common under septic conditions. Skeletal muscle is the main target organ for hypercatabolism, and this phenomenon is a vital factor in the deterioration of recovery in septic patients. In skeletal muscle, activation of the ubiquitin-proteasome system plays an important role in hypercatabolism under septic status. Insulin is a vital anticatabolic hormone and previous evidence suggests that insulin administration inhibits various steps in the ubiquitin proteasome system. However, whether insulin can alleviate the degradation of skeletal muscle protein by inhibiting the ubiquitin-proteasome system under septic condition is unclear. This paper confirmed that mRNA and protein levels of the ubiquitin-proteasome system were upregulated and molecular markers of skeletal muscle proteolysis (tyrosine and 3-methylhistidine) simultaneously increased in the skeletal muscle of septic rats. Septic rats were infused with insulin at a constant rate of 2.4 mU.kg-1.min-1 for 8 hours. Concentrations of mRNA and proteins of the ubiquitin-proteasome system and molecular markers of skeletal muscle proteolysis were mildly affected. When the insulin infusion dose increased to 4.8 mU.kg-1.min-1, mRNA for ubiquitin, E2-14 KDa, and the C2 subunit were all sharply downregulated. At the same time, the levels of ubiquitinated proteins, E2-14KDa, and the C2 subunit protein were significantly reduced. Tyrosine and 3-methylhistidine decreased significantly. We concluded that the ubiquitin-proteasome system is important skeletal muscle hypercatabolism in septic rats. Infusion of insulin can reverse the detrimental metabolism of skeletal muscle by inhibiting the ubiquitin-proteasome system, and the effect is proportional to the insulin infusion dose. PMID- 21639906 TI - Do changes in traditional coronary heart disease risk factors over time explain the association between socio-economic status and coronary heart disease? AB - BACKGROUND: Socioeconomic status (SES) predicts coronary heart disease independently of the traditional risk factors included in the Framingham risk score. However, it is unknown whether changes in Framingham risk score variables over time explain the association between SES and coronary heart disease. We examined this question given its relevance to risk assessment in clinical decision making. METHODS: The Atherosclerosis Risk in Communities study data (initiated in 1987 with 10-years follow-up of 15,495 adults aged 45-64 years in four Southern and Mid-Western communities) were used. SES was assessed at baseline, dichotomized as low SES (defined as low education and/or low income) or not. The time dependent variables - smoking, total and high density lipoprotein cholesterol, systolic blood pressure and use of blood pressure lowering medication - were assessed every three years. Ten-year incidence of coronary heart disease was based on EKG and cardiac enzyme criteria, or adjudicated death certificate data. Cox survival analyses examined the contribution of SES to heart disease risk independent of baseline Framingham risk score, without and with further adjustment for the time dependent variables. RESULTS: Adjusting for baseline Framingham risk score, low SES was associated with an increased coronary heart disease risk (hazard ratio [HR] = 1.53; 95% Confidence Interval [CI], 1.27 to 1.85). After further adjustment for the time dependent variables, the SES effect remained significant (HR = 1.44; 95% CI, 1.19 to 1.74). CONCLUSION: Using Framingham Risk Score alone under estimated the coronary heart disease risk in low SES persons. This bias was not eliminated by subsequent changes in Framingham risk score variables. PMID- 21639907 TI - Infliximab in ankylosing spondylitis: alone or in combination with methotrexate? A pharmacokinetic comparative study. AB - INTRODUCTION: Methotrexate (MTX) has been shown to modify infliximab pharmacokinetics in rheumatoid arthritis. However, its combination with infliximab in the treatment of ankylosing spondylitis (AS) is not recommended. The objective of this study was to examine the influence of MTX on infliximab exposure in patients with AS. METHODS: Patients with AS patients who had predominantly axial symptoms were randomised to receive infliximab alone (infusions of 5 mg/kg at weeks 0, 2, 6, 12 and 18) or infliximab combined with MTX (10 mg/week). Infliximab concentrations were measured before and 2 hours after each infusion and at 1, 3, 4, 5, 8, 10, 14 and 18 weeks. We estimated individual cumulative area under the concentration versus time curves (AUC) for infliximab concentration between baseline and week 18 (AUC(0-18)). Clinical and laboratory evaluations were performed at each visit. The Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) score was the primary end point for clinical response. RESULTS: Twenty-six patients were included (infliximab group: n = 12, infliximab + MTX group: n = 14), and 507 serum samples were available for measurement of infliximab concentration. The two groups did not differ with regard to AUC(0-18) or evolution of BASDAI scores and biomarkers of inflammation. CONCLUSIONS: The combination of MTX and infliximab does not increase the exposure to infliximab over infliximab alone in patients with AS. TRIAL REGISTRATION: ClinicalTrials.gov: NCT00507403. PMID- 21639908 TI - A honey bee (Apis mellifera L.) PeptideAtlas crossing castes and tissues. AB - BACKGROUND: Honey bees are a mainstay of agriculture, contributing billions of dollars through their pollination activities. Bees have been a model system for sociality and group behavior for decades but only recently have molecular techniques been brought to study this fascinating and valuable organism. With the release of the first draft of its genome in 2006, proteomics of bees became feasible and over the past five years we have amassed in excess of 5E+6 MS/MS spectra. The lack of a consolidated platform to organize this massive resource hampers our ability, and that of others, to mine the information to its maximum potential. RESULTS: Here we introduce the Honey Bee PeptideAtlas, a web-based resource for visualizing mass spectrometry data across experiments, providing protein descriptions and Gene Ontology annotations where possible. We anticipate that this will be helpful in planning proteomics experiments, especially in the selection of transitions for selected reaction monitoring. Through a proteogenomics effort, we have used MS/MS data to anchor the annotation of previously undescribed genes and to re-annotate previous gene models in order to improve the current genome annotation. CONCLUSIONS: The Honey Bee PeptideAtlas will contribute to the efficiency of bee proteomics and accelerate our understanding of this species. This publicly accessible and interactive database is an important framework for the current and future analysis of mass spectrometry data. PMID- 21639910 TI - Work-related pesticide poisoning among farmers in two villages of Southern China: a cross-sectional survey. AB - BACKGROUND: Pesticide poisoning is an important health problem among Chinese farm workers, but there is a paucity of pesticide poisoning data from China. Using the WHO standard case definition of a possible acute pesticide poisoning, we investigated the prevalence and risk factors of acute work-related pesticide poisoning among farmers in Southern China. METHODS: A stratified sample of 910 pesticide applicators from two villages in southern China participated in face-to face interviews. Respondents who self-reported having two or more of a list of sixty-six symptoms within 24 hours after pesticide application were categorized as having suffered acute pesticide poisoning. The association between the composite behavioral risk score and pesticide poisoning were assessed in a multivariate logistic model. RESULTS: A total of 80 (8.8%) pesticide applicators reported an acute work-related pesticide poisoning. The most frequent symptoms among applicators were dermal (11.6%) and nervous system (10.7%) symptoms. Poisoning was more common among women, farmers in poor areas, and applicators without safety training (all p < 0.001). After controlling for gender, age, education, geographic area and the behavioral risk score, farmers without safety training had an adjusted odds ratio of 3.22 (95% CI: 1.86-5.60). The likelihood of acute pesticide poisoning was also significantly associated with number of exposure risk behaviors. A significant "dose-response" relationship between composite behavioral risk scores calculated from 9 pesticides exposure risk behaviors and the log odds of pesticide poisoning prevalence was seen among these Chinese farmers (R2 = 0.9246). CONCLUSIONS: This study found that 8.8% of Chinese pesticide applicators suffered acute pesticide poisoning and suggests that pesticide safety training, safe application methods, and precautionary behavioral measures could be effective in reducing the risk of pesticide poisoning. PMID- 21639909 TI - The role of female hormones on lung function in chronic lung diseases. AB - BACKGROUND: The prevalence, morbidity, and mortality of inflammatory lung diseases such as asthma, chronic obstructive pulmonary disease (COPD) and cystic fibrosis (CF) are increasing in women. There is a dearth of data on the biological mechanisms to explain such observations. However, some large epidemiologic studies suggest that lung function fluctuates during the menstrual cycle in female patients with airways disease but not in women without disease, suggesting that circulating estradiol and progesterone may be involved in this process. DISCUSSION: In asthma, estradiol shuttles adaptive immunity towards the TH2 phenotype while in smokers estrogens may be involved in the generation of toxic intermediate metabolites in the airways of female smokers, which may be relevant in COPD pathogenesis. In CF, estradiol has been demonstrated to up regulate MUC5B gene in human airway epithelial cells and inhibit chloride secretion in the airways. Progesterone may augment airway inflammation. SUMMARY: Taken together, clinical and in-vivo data have demonstrated a sex-related difference in that females may be more susceptible to the pathogenesis of lung diseases. In this paper, we review the effect of female sex hormones in the context of these inflammatory airway diseases. PMID- 21639911 TI - Mouldy feed, mycotoxins and Shiga toxin - producing Escherichia coli colonization associated with Jejunal Hemorrhage Syndrome in beef cattle. AB - BACKGROUND: Both O157 and non-O157 Shiga toxin - producing Escherichia coli (STECs) cause serious human disease outbreaks through the consumption of contaminated foods. Cattle are considered the main reservoir but it is unclear how STECs affect mature animals. Neonatal calves are the susceptible age class for STEC infections causing severe enteritis. In an earlier study, we determined that mycotoxins and STECs were part of the disease complex for dairy cattle with Jejunal Hemorrhage Syndrome (JHS). For STECs to play a role in the development of JHS, we hypothesized that STEC colonization should also be evident in beef cattle with JHS. Aggressive medical and surgical therapies are effective for JHS, but rely on early recognition of clinical signs for optimal outcomes suggesting that novel approaches must be developed for managing this disease. The main objective of this study was to confirm that mouldy feeds, mycotoxins and STEC colonization were associated with the development of JHS in beef cattle. RESULTS: Beef cattle developed JHS after consuming feed containing several types of mycotoxigenic fungi including Fusarium poae, F. verticillioides, F. sporotrichioides, Penicillium roqueforti and Aspergillus fumigatus. Mixtures of STECs colonized the mucosa in the hemorrhaged tissues of the cattle and no other pathogen was identified. The STECs expressed Stx1 and Stx2, but more significantly, Stxs were also present in the blood collected from the lumen of the hemorrhaged jejunum. Feed extracts containing mycotoxins were toxic to enterocytes and 0.1% of a prebiotic, Celmanax Trademark, removed the cytotoxicity in vitro. The inclusion of a prebiotic in the care program for symptomatic beef calves was associated with 69% recovery. CONCLUSIONS: The current study confirmed that STECs and mycotoxins are part of the disease complex for JHS in beef cattle. Mycotoxigenic fungi are only relevant in that they produce the mycotoxins deposited in the feed. A prebiotic, Celmanax Trademark, acted as a mycotoxin binder in vitro and interfered with the progression of disease. PMID- 21639912 TI - HAPIscreen, a method for high-throughput aptamer identification. AB - BACKGROUND: Aptamers are oligonucleotides displaying specific binding properties for a predetermined target. They are selected from libraries of randomly synthesized candidates through an in vitro selection process termed SELEX (Systematic Evolution of Ligands by EXponential enrichment) alternating selection and amplification steps. SELEX is followed by cloning and sequencing of the enriched pool of oligonucleotides to enable comparison of the selected sequences. The most represented candidates are then synthesized and their binding properties are individually evaluated thus leading to the identification of aptamers. These post-selection steps are time consuming and introduce a bias to the expense of poorly amplified binders that might be of high affinity and are consequently underrepresented. A method that would circumvent these limitations would be highly valuable. RESULTS: We describe a novel homogeneous solution-based method for screening large populations of oligonucleotide candidates generated from SELEX. This approach, based on the AlphaScreen(r) technology, is carried out on the exclusive basis of the binding properties of the selected candidates without the needs of performing a priori sequencing. It therefore enables the functional identification of high affinity aptamers. We validated the HAPIscreen (High throughput APtamer Identification screen) methodology using aptamers targeted to RNA hairpins, previously identified in our laboratory. We then screened pools of candidates issued from SELEX rounds in a 384 well microplate format and identify new RNA aptamers to pre-microRNAs. CONCLUSIONS: HAPIscreen, an Alphascreen(r) based methodology for the identification of aptamers is faster and less biased than current procedures based on sequence comparison of selected oligonucleotides and sampling binding properties of few individuals. Moreover this methodology allows for screening larger number of candidates. Used here for selecting anti premiR aptamers, HAPIscreen can be adapted to any type of tagged target and is fully amenable to automation. PMID- 21639913 TI - Transcriptomics and molecular evolutionary rate analysis of the bladderwort (Utricularia), a carnivorous plant with a minimal genome. AB - BACKGROUND: The carnivorous plant Utricularia gibba (bladderwort) is remarkable in having a minute genome, which at ca. 80 megabases is approximately half that of Arabidopsis. Bladderworts show an incredible diversity of forms surrounding a defined theme: tiny, bladder-like suction traps on terrestrial, epiphytic, or aquatic plants with a diversity of unusual vegetative forms. Utricularia plants, which are rootless, are also anomalous in physiological features (respiration and carbon distribution), and highly enhanced molecular evolutionary rates in chloroplast, mitochondrial and nuclear ribosomal sequences. Despite great interest in the genus, no genomic resources exist for Utricularia, and the substitution rate increase has received limited study. RESULTS: Here we describe the sequencing and analysis of the Utricularia gibba transcriptome. Three different organs were surveyed, the traps, the vegetative shoot bodies, and the inflorescence stems. We also examined the bladderwort transcriptome under diverse stress conditions. We detail aspects of functional classification, tissue similarity, nitrogen and phosphorus metabolism, respiration, DNA repair, and detoxification of reactive oxygen species (ROS). Long contigs of plastid and mitochondrial genomes, as well as sequences for 100 individual nuclear genes, were compared with those of other plants to better establish information on molecular evolutionary rates. CONCLUSION: The Utricularia transcriptome provides a detailed genomic window into processes occurring in a carnivorous plant. It contains a deep representation of the complex metabolic pathways that characterize a putative minimal plant genome, permitting its use as a source of genomic information to explore the structural, functional, and evolutionary diversity of the genus. Vegetative shoots and traps are the most similar organs by functional classification of their transcriptome, the traps expressing hydrolytic enzymes for prey digestion that were previously thought to be encoded by bacteria. Supporting physiological data, global gene expression analysis shows that traps significantly over-express genes involved in respiration and that phosphate uptake might occur mainly in traps, whereas nitrogen uptake could in part take place in vegetative parts. Expression of DNA repair and ROS detoxification enzymes may be indicative of a response to increased respiration. Finally, evidence from the bladderwort transcriptome, direct measurement of ROS in situ, and cross-species comparisons of organellar genomes and multiple nuclear genes supports the hypothesis that increased nucleotide substitution rates throughout the plant may be due to the mutagenic action of amplified ROS production. PMID- 21639914 TI - Premenstrual symptoms in young adults born preterm at very low birth weight--from the Helsinki Study of Very Low Birth Weight Adults. AB - BACKGROUND: Clinically significant premenstrual symptoms are common among young women. Premenstrual syndrome (PMS) is characterized by emotional, behavioural and physical symptoms that consistently occur during the luteal phase of the menstrual cycle. Premenstrual dysphoric disorder (PMDD) is a severe form of PMS. Individual variation in stress responsiveness may be involved in the pathophysiology of premenstrual symptoms. Preterm birth at very low birth weight (VLBW, < 1500g) has a multitude of consequences that extend to adult life, including altered stress responsiveness which could affect the prevalence of premenstrual symptoms. METHODS: In this cohort study, we compared 75 VLBW women with 95 women born at term (mean age 22.5). We used a standardized retrospective questionnaire assessing the presence and severity of a variety of symptoms before and after menses. The symptom scores were used both as continuous and as dichotomized variables, with cutoffs based on DSM-IV criteria for PMDD and ACOG criteria for PMS, except prospective daily ratings could not be used. We used multiple linear and logistic regression to adjust for confounders. RESULTS: There was no difference in the continuous symptom score before menses (mean difference VLBW-term -18.3%, 95% confidence interval -37.9 to 7.5%) or after menses. The prevalence of premenstrual symptoms causing severe impairment to daily life was 13.3% for VLBW women and 14.7% for control women. For PMDD, it was 8.0% and 4.2%, and for PMS, 12.0% and 11.6%, respectively. These differences were not statistically significant (p > 0.1). CONCLUSION: Our findings suggest that the severity of premenstrual symptoms and the prevalence of PMDD and PMS among young women born preterm at VLBW is not higher than among those born at term. PMID- 21639915 TI - Genistein inhibits proliferation of colon cancer cells by attenuating a negative effect of epidermal growth factor on tumor suppressor FOXO3 activity. AB - BACKGROUND: Soy consumption is associated with a lower incidence of colon cancer which is believed to be mediated by one of its of components, genistein. Genistein may inhibit cancer progression by inducing apoptosis or inhibiting proliferation, but mechanisms are not well understood. Epidermal growth factor (EGF)-induced proliferation of colon cancer cells plays an important role in colon cancer progression and is mediated by loss of tumor suppressor FOXO3 activity. The aim of this study was to assess if genistein exerts anti proliferative properties by attenuating the negative effect of EGF on FOXO3 activity. METHODS: The effect of genistein on proliferation stimulated by EGF mediated loss of FOXO3 was examined in human colonic cancer HT-29 cells. EGF induced FOXO3 phosphorylation and translocation were assessed in the presence of genistein. EGF-mediated loss of FOXO3 interactions with p53 (co immunoprecipitation) and promoter of p27kip1 (ChIP assay) were examined in presence of genistein in cells with mutated p53 (HT-29) and wild type p53 (HCT116). Silencing of p53 determined activity of FOXO3 when it is bound to p53. RESULTS: Genistein inhibited EGF-induced proliferation, while favoring dephosphorylation and nuclear retention of FOXO3 (active state) in colon cancer cells. Upstream of FOXO3, genistein acts via the PI3K/Akt pathway to inhibit EGF stimulated FOXO3 phosphorylation (i.e. favors active state). Downstream, EGF induced disassociation of FOXO3 from mutated tumor suppressor p53, but not wild type p53, is inhibited by genistein favoring FOXO3-p53(mut) interactions with the promoter of the cell cycle inhibitor p27kip1 in colon cancer cells. Thus, the FOXO3-p53(mut) complex leads to elevated p27kip1 expression and promotes cell cycle arrest. CONCLUSION: These novel anti-proliferative mechanisms of genistein suggest a possible role of combining genistein with other chemoreceptive agents for the treatment of colon cancer. PMID- 21639916 TI - Reduction of central venous catheter associated blood stream infections following implementation of a resident oversight and credentialing policy. AB - BACKGROUND: This study assesses the impact that a resident oversight and credentialing policy for central venous catheter (CVC) placement had on institution-wide central line associated bloodstream infections (CLABSI). We therefore investigated the rate of CLABSI per 1,000 line days during the 12 months before and after implementation of the policy. METHODS: This is a retrospective analysis of prospectively collected data at an academic medical center with four adult ICUs and a pediatric ICU. All patients undergoing non tunneled CVC placement were included in the study. Data was collected on CLABSI, line days, and serious adverse events in the year prior to and following policy implementation on 9/01/08. RESULTS: A total of 813 supervised central lines were self-reported by residents in four departments. Statistical analysis was performed using paired Wilcoxon signed rank tests. There were reductions in median CLABSI rate (3.52 vs. 2.26; p = 0.015), number of CLBSI per month (16.0 to 10.0; p = 0.012), and line days (4495 vs. 4193; p = 0.019). No serious adverse events reported to the Pennsylvania Patient Safety Authority. CONCLUSIONS: Implementation of a new CVC resident oversight and credentialing policy has been significantly associated with an institution-wide reduction in the rate of CLABSI per 1,000 central line days and total central line days. No serious adverse events were reported. Similar resident oversight policies may benefit other teaching institutions, and support concurrent organizational efforts to reduce hospital acquired infections. PMID- 21639917 TI - Momordica charantia (bitter melon) attenuates high-fat diet-associated oxidative stress and neuroinflammation. AB - BACKGROUND: The rising epidemic of obesity is associated with cognitive decline and is considered as one of the major risk factors for neurodegenerative diseases. Neuroinflammation is a critical component in the progression of several neurological and neurodegenerative diseases. Increased metabolic flux to the brain during overnutrition and obesity can orchestrate stress response, blood brain barrier (BBB) disruption, recruitment of inflammatory immune cells from peripheral blood and microglial cells activation leading to neuroinflammation. The lack of an effective treatment for obesity-associated brain dysfunction may have far-reaching public health ramifications, urgently necessitating the identification of appropriate preventive and therapeutic strategies. The objective of our study was to investigate the neuroprotective effects of Momordica charantia (bitter melon) on high-fat diet (HFD)-associated BBB disruption, stress and neuroinflammatory cytokines. METHODS: C57BL/6 female mice were fed HFD with and without bitter melon (BM) for 16 weeks. BBB disruption was analyzed using Evans blue dye. Phosphate-buffered saline (PBS) perfused brains were analyzed for neuroinflammatory markers such as interleukin-22 (IL-22), IL 17R, IL-16, NF-kappaB1, and glial cells activation markers such as Iba1, CD11b, GFAP and S100beta. Additionally, antioxidant enzymes, ER-stress proteins, and stress-resistant transcription factors, sirtuin 1 (Sirt1) and forkhead box class O transcription factor (FoxO) were analyzed using microarray, quantitative real time RT-PCR, western immunoblotting and enzymatic assays. Systemic inflammation was analyzed using cytokine antibody array. RESULTS: BM ameliorated HFD associated changes in BBB permeability as evident by reduced leakage of Evans blue dye. HFD-induced glial cells activation and expression of neuroinflammatory markers such as NF-kappaB1, IL-16, IL-22 as well as IL-17R were normalized in the brains of mice supplemented with BM. Similarly, HFD-induced brain oxidative stress was significantly reduced by BM supplementation with a concomitant reduction in FoxO, normalization of Sirt1 protein expression and up-regulation of Sirt3 mRNA expression. Furthermore, plasma antioxidant enzymes and pro inflammatory cytokines were also normalized in mice fed HFD with BM as compared to HFD-fed mice. CONCLUSIONS: Functional foods such as BM offer a unique therapeutic strategy to improve obesity-associated peripheral inflammation and neuroinflammation. PMID- 21639919 TI - Mucopolysaccharidosis type I: molecular characteristics of two novel alpha-L iduronidase mutations in Tunisian patients. AB - BACKGROUND: Mucopolysaccharidosis type I (MPS I) is an autosomal storage disease resulting from defective activity of the enzyme alpha-L-iduronidase (IDUA). This glycosidase is involved in the degradation of heparan sulfate and dermatan sulfate. MPS I has severe and milder phenotypic subtypes. AIM OF STUDY: This study was carried out on six newly collected MPS I patients recruited from many regions of Tunisia. PATIENTS AND METHODS: Mutational analysis of the IDUA gene in unrelated MPS I families was performed by sequencing the exons and intron-exon junctions of IDUA gene. RESULTS: Two novel IDUA mutations, p.L530fs (1587_1588 insGC) in exon 11 and p.F177S in exon 5 and two previously reported mutations p.P533R and p.Y581X were detected. The patient in family 1 who has the Hurler phenotype was homozygous for the previously described nonsense mutation p.Y581X.The patient in family 2 who also has the Hurler phenotype was homozygous for the novel missense mutation p.F177S. The three patients in families 3, 5 and 6 were homozygous for the p.P533R mutation. The patient in family 4 was homozygous for the novel small insertion 1587_1588 insGC. In addition, eighteen known and one unknown IDUA polymorphisms were identified. CONCLUSION: The identification of these mutations should facilitate prenatal diagnosis and counseling for MPS I in Tunisia. PMID- 21639918 TI - Interpopulation hybridization results in widespread viability selection across the genome in Tigriopus californicus. AB - BACKGROUND: Genetic interactions within hybrids influence their overall fitness. Understanding the details of these interactions can improve our understanding of speciation. One experimental approach is to investigate deviations from Mendelian expectations (segregation distortion) in the inheritance of mapped genetic markers. In this study, we used the copepod Tigriopus californicus, a species which exhibits high genetic divergence between populations and a general pattern of reduced fitness in F2 interpopulation hybrids. Previous studies have implicated both nuclear-cytoplasmic and nuclear-nuclear interactions in causing this fitness reduction. We identified and mapped population-diagnostic single nucleotide polymorphisms (SNPs) and used these to examine segregation distortion across the genome within F2 hybrids. RESULTS: We generated a linkage map which included 45 newly elucidated SNPs and 8 population-diagnostic microsatellites used in previous studies. The map, the first available for the Copepoda, was estimated to cover 75% of the genome and included markers on all 12 T. californicus chromosomes. We observed little segregation distortion in newly hatched F2 hybrid larvae (fewer than 10% of markers at p < 0.05), but strikingly higher distortion in F2 hybrid adult males (45% of markers at p < 0.05). Hence, segregation distortion was primarily caused by selection against particular genetic combinations which acted between hatching and maturity. Distorted markers were not distributed randomly across the genome but clustered on particular chromosomes. In contrast to other studies in this species we found little evidence for cytonuclear coadaptation. Instead, different linkage groups exhibited markedly different patterns of distortion, which appear to have been influenced by nuclear-nuclear epistatic interactions and may also reflect genetic load carried within the parental lines. CONCLUSION: Adult male F2 hybrids between two populations of T. californius exhibit dramatic segregation distortion across the genome. Distorted loci are clustered within specific linkage groups, and the direction of distortion differs between chromosomes. This segregation distortion is due to selection acting between hatching and adulthood. PMID- 21639920 TI - Genomic comparison of multi-drug resistant invasive and colonizing Acinetobacter baumannii isolated from diverse human body sites reveals genomic plasticity. AB - BACKGROUND: Acinetobacter baumannii has recently emerged as a significant global pathogen, with a surprisingly rapid acquisition of antibiotic resistance and spread within hospitals and health care institutions. This study examines the genomic content of three A. baumannii strains isolated from distinct body sites. Isolates from blood, peri-anal, and wound sources were examined in an attempt to identify genetic features that could be correlated to each isolation source. RESULTS: Pulsed-field gel electrophoresis, multi-locus sequence typing and antibiotic resistance profiles demonstrated genotypic and phenotypic variation. Each isolate was sequenced to high-quality draft status, which allowed for comparative genomic analyses with existing A. baumannii genomes. A high resolution, whole genome alignment method detailed the phylogenetic relationships of sequenced A. baumannii and found no correlation between phylogeny and body site of isolation. This method identified genomic regions unique to both those isolates found on the surface of the skin or in wounds, termed colonization isolates, and those identified from body fluids, termed invasive isolates; these regions may play a role in the pathogenesis and spread of this important pathogen. A PCR-based screen of 74 A. baumanii isolates demonstrated that these unique genes are not exclusive to either phenotype or isolation source; however, a conserved genomic region exclusive to all sequenced A. baumannii was identified and verified. CONCLUSIONS: The results of the comparative genome analysis and PCR assay show that A. baumannii is a diverse and genomically variable pathogen that appears to have the potential to cause a range of human disease regardless of the isolation source. PMID- 21639921 TI - Association of promoter methylation with histologic type and pleural indentation in non-small cell lung cancer (NSCLC). AB - BACKGROUND: Lung cancer is a major cause of death worldwide. Gene promoter methylation is a major inactivation mechanism of tumor-related genes, some of which can be served as a biomarker for early diagnosis and prognosis evaluation of lung cancer. METHODS: We determined the promoter methylation of 6 genes using quantitative methylation-specific PCR (Q-MSP) technique in 96 clinically well characterized non-small cell lung cancer (NSCLC). RESULTS: Highly frequent promoter methylation was found in NSCLC. With 100% diagnostic specificity, high sensitivity, ranging from 44.9 to 84.1%, was found for each of the 6 genes. Our data also showed that promoter methylation was closely associated with histologic type. Most of genes were more frequently methylated in squamous cell carcinomas (SCC) compared to adenocarcinomas (ADC). Moreover, promoter methylation significantly increased the risk of pleural indentation in NSCLC. CONCLUSION: Our findings provided evidences that multiple genes were aberrantly methylated in lung tumorigenesis, and demonstrated the promoter methylation was closely associated with clinicopathologic characteristics of NSCLC. More importantly, we first revealed promoter methylation may be served as a potentially increased risk factor for pleural indentation of NSCLC patients. PMID- 21639922 TI - Involvement of the Cav3.2 T-type calcium channel in thalamic neuron discharge patterns. AB - BACKGROUND: Mice that have defects in their low-threshold T-type calcium channel (T-channel) genes show altered pain behaviors. The changes in the ratio of nociceptive neurons and the burst firing property of reticular thalamic (RT) and ventroposterior (VP) neurons in Cav3.2 knockout (KO) mice were studied to test the involvement of thalamic T-channel and burst firing activity in pain function. RESULTS: Under pentobarbital or urethane anesthesia, the patterns of tonic and burst firings were recorded in functionally characterized RT and VPL neurons of Cav3.2 KO mice. Many RT neurons were nociceptive (64% under pentobarbital anesthesia and 50% under urethane anesthesia). Compared to their wild-type (WT) controls, fewer nociceptive RT neurons were found in Cav3.2 KO mice. Both nociceptive and tactile RT neurons showed fewer bursts in Cav3.2 KO mice. Within a burst, RT neurons of Cav3.2 KO mice had a lower spike frequency and less prominent accelerando-decelerando change. In contrast, VP neurons of Cav3.2 KO mice showed a higher ratio of bursts and a higher discharge rate within a burst than those of the WT control. In addition, the long-lasting tonic firing episodes in RT neurons of the Cav3.2 KO had less stereotypic regularity than their counterparts in WT mice. CONCLUSIONS: RT might be important in nociception of the mouse. In addition, we showed an important role of Cav3.2 subtype of T-channel in RT burst firing pattern. The decreased occurrence and slowing of the bursts in RT neurons might cause the increased VP bursts. These changes would be factors contributing to alternation of pain behavior in the Cav3.2 KO mice. PMID- 21639923 TI - A traditional Chinese medicine versus Western combination therapy in the treatment of rheumatoid arthritis: two-stage study protocol for a randomized controlled trial. AB - BACKGROUND: The common randomized controlled trial design has distinct limitations when applied to Chinese medicine, because Chinese medicine identifies and treats 'Chinese medicine patterns' rather than diagnosed diseases. Chinese medicine patterns are a group of associated symptoms, tongue appearances and pulse characteristics. These limitations could be overcome by developing new strategies to evaluate the effect of Chinese medicine. The idea behind pattern based efficacy evaluations may optimize clinical trial design by identifying the responsiveness-related Chinese medicine patterns. METHODS/DESIGN: This is a two stage multi-center trial of Chinese herbal medicine for the management of rheumatoid arthritis. The stage one trial is an open-label trial and aims to explore what groups of Chinese medicine information (such as symptoms) correlates with better efficacy, and the stage two trial is a randomized, controlled, double blind, double-dummy clinical trial that incorporates the efficacy-related information identified in the stage-one trial into the inclusion criteria. DISCUSSION: The indication of a Chinese herbal formula is a specific Chinese medicine pattern and not a single disease and stratifying a disease into several patterns with a group of symptoms is a feasible procedure in clinical trials. This study is the first to investigate whether this approach in the design of Chinese herbal medicine trials can improve responses. TRIAL REGISTRATION: ChiCTR TRC-10000989. PMID- 21639924 TI - Surgical and conservative treatment of patients with congenital scoliosis: alpha search for long-term results. AB - BACKGROUND: In view of the limited data available on the conservative treatment of patients with congenital scoliosis (CS), early surgery is suggested in mild cases with formation failures. Patients with segmentation failures will not benefit from conservative treatment. The purpose of this review is to identify the mid- or long-term results of spinal fusion surgery in patients with congenital scoliosis. METHODS: Retrospective and prospective studies were included, reporting on the outcome of surgery in patients with congenital scoliosis. Studies concerning a small numbers of cases treated conservatively were included too. We analyzed mid-term (5 to 7 years) and long-term results (7 years or more), both as regards the maintenance of the correction of scoliosis and the safety of instrumentation, the early and late complications of surgery and their effect on quality of life. RESULTS: A small number of studies of surgically treated patients were found, contained follow-up periods of 4-6 years that in the most cases, skeletal maturity was not yet reached, and few with follow-up of 36-44 years. The results of bracing in children with congenital scoliosis, mainly in cases with failure of formation, were also studied. DISCUSSION: Spinal surgery in patients with congenital scoliosis is regarded in short as a safe procedure and should be performed. On the other hand, early and late complications are also described, concerning not only intraoperative and immediate postoperative problems, but also the safety and efficacy of the spinal instrumentation and the possibility of developing neurological disorders and the long-term effect these may have on both lung function and the quality of life of children. CONCLUSIONS: Few cases indicate the long-term results of surgical techniques, in the natural progression of scoliosis. Similarly, few cases have been reported on the influence of conservative treatment.In conclusion, patients with segmentation failures should be treated surgically early, according to the rate of deformity formation and certainly before the pubertal growth spurt to try to avoid cor- pulmonale, even though there is lack of evidence for that in the long-term. Furthermore, in patients with formation failures, further investigation is needed to document where a conservative approach would be necessary. PMID- 21639925 TI - Growth, physicochemical properties, and morphogenesis of Chinese wild-type PRV Fa and its gene-deleted mutant strain PRV SA215. AB - BACKGROUND: PRV Fa is common in China and causes most of the pseudorabies in the pig industry. A PRV SA215 strain with deleted gE, gI, and TK genes was constructed to develop a commercial attenuated live vaccine. However, the physicochemical properties, growth pattern, penetration kinetics, and morphogenesis of the PRV SA215 and its parental PRV Fa strain are unclear. RESULTS: A series of experiments were conducted to characterize both strains and provide more information. PRV Fa and PRV SA215 were found to have similar penetration patterns, with about 5 min half-time of penetration. The SA215 strain exhibited a slight delay in entry compared with PRV Fa. In the one-step growth test, the titers of the SA215 strain were first detected at 8 h, rapidly increased, and peaked at 12 h. A plateau was formed between 12-36 h of culturing. PRV SA215 showed delayed replication and approximately 10-30-fold lower titers during 0-16 h of culturing compared with the PRV-Fa strain. After 16 h, the PRV Fa titers dramatically decreased, whereas those of PRV SA215 were prolonged to 36 h and reached a titer value equal to that of PRV Fa and then decreased. Both strains were sensitive to both heat and acid-alkali treatments; however, PRV Fa was relatively more stable to heat treatment than PRV SA215. Both strains could propagate in the cultures with pH values from 5.0 to 9.0. Cultures with pH below 3.0 or above 11.0 were fatal to both strains. Both strains had considerable resistance to freeze-thawing treatments. Morphogenetic investigations showed that typical phases in the maturation pathway were observed in the PRV Fa-infected PK15 cells, whereas secondary envelopment was not observed in the PRV SA215 strain. Instead, capsid aggregations with concomitants of electrodense materials were observed. CONCLUSIONS: These results suggest that PRV SA215 is a promising strain for vaccine development. PMID- 21639926 TI - Malaria in Kakuma refugee camp, Turkana, Kenya: facilitation of Anopheles arabiensis vector populations by installed water distribution and catchment systems. AB - BACKGROUND: Malaria is a major health concern for displaced persons occupying refugee camps in sub-Saharan Africa, yet there is little information on the incidence of infection and nature of transmission in these settings. Kakuma Refugee Camp, located in a dry area of north-western Kenya, has hosted ca. 60,000 to 90,000 refugees since 1992, primarily from Sudan and Somalia. The purpose of this study was to investigate malaria prevalence and attack rate and sources of Anopheles vectors in Kakuma refugee camp, in 2005-2006, after a malaria epidemic was observed by staff at camp clinics. METHODS: Malaria prevalence and attack rate was estimated from cases of fever presenting to camp clinics and the hospital in August 2005, using rapid diagnostic tests and microscopy of blood smears. Larval habitats of vectors were sampled and mapped. Houses were sampled for adult vectors using the pyrethrum knockdown spray method, and mapped. Vectors were identified to species level and their infection with Plasmodium falciparum determined. RESULTS: Prevalence of febrile illness with P. falciparum was highest among the 5 to 17 year olds (62.4%) while malaria attack rate was highest among the two to 4 year olds (5.2/1,000/day). Infected individuals were spatially concentrated in three of the 11 residential zones of the camp. The indoor densities of Anopheles arabiensis, the sole malaria vector, were similar during the wet and dry seasons, but were distributed in an aggregated fashion and predominantly in the same zones where malaria attack rates were high. Larval habitats and larval populations were also concentrated in these zones. Larval habitats were man-made pits of water associated with tap-stands installed as the water delivery system to residents with year round availability in the camp. Three percent of A. arabiensis adult females were infected with P. falciparum sporozoites in the rainy season. CONCLUSIONS: Malaria in Kakuma refugee camp was due mainly to infection with P. falciparum and showed a hyperendemic age prevalence profile, in an area with otherwise low risk of malaria given prevailing climate. Transmission was sustained by A. arabiensis, whose populations were facilitated by installation of man-made water distribution and catchment systems. PMID- 21639927 TI - Pilot randomized trial of therapeutic hypothermia with serial cranial ultrasound and 18-22 month follow-up for neonatal encephalopathy in a low resource hospital setting in Uganda: study protocol. AB - BACKGROUND: There is now convincing evidence that in industrialized countries therapeutic hypothermia for perinatal asphyxial encephalopathy increases survival with normal neurological function. However, the greatest burden of perinatal asphyxia falls in low and mid-resource settings where it is unclear whether therapeutic hypothermia is safe and effective. AIMS: Under the UCL Uganda Women's Health Initiative, a pilot randomized controlled trial in infants with perinatal asphyxia was set up in the special care baby unit in Mulago Hospital, a large public hospital with ~20,000 births in Kampala, Uganda to determine:(i) The feasibility of achieving consent, neurological assessment, randomization and whole body cooling to a core temperature 33-34 degrees C using water bottles(ii) The temperature profile of encephalopathic infants with standard care(iii) The pattern, severity and evolution of brain tissue injury as seen on cranial ultrasound and relation with outcome(iv) The feasibility of neurodevelopmental follow-up at 18-22 months of age METHODS/DESIGN: Ethical approval was obtained from Makerere University and Mulago Hospital. All infants were in-born. Parental consent for entry into the trial was obtained. Thirty-six infants were randomized either to standard care plus cooling (target rectal temperature of 33-34 degrees C for 72 hrs, started within 3 h of birth) or standard care alone. All other aspects of management were the same. Cooling was performed using water bottles filled with tepid tap water (25 degrees C). Rectal, axillary, ambient and surface water bottle temperatures were monitored continuously for the first 80 h. Encephalopathy scoring was performed on days 1-4, a structured, scorable neurological examination and head circumference were performed on days 7 and 17. Cranial ultrasound was performed on days 1, 3 and 7 and scored. Griffiths developmental quotient, head circumference, neurological examination and assessment of gross motor function were obtained at 18-22 months. DISCUSSION: We will highlight differences in neonatal care and infrastructure that need to be taken into account when considering a large safety and efficacy RCT of therapeutic hypothermia in low and mid resource settings in the future. TRIAL REGISTRATION: Current controlled trials ISRCTN92213707. PMID- 21639928 TI - Successful bone marrow transplantation in a patient with Diamond-Blackfan anemia with co-existing Duchenne muscular dystrophy: a case report. AB - INTRODUCTION: Diamond-Blackfan anemia and Duchenne muscular dystrophy are two rare congenital anomalies. Both anomalies occurring in the same child is extremely rare. Allogeneic hematopoietic stem cell transplantation is a well established therapy for Diamond-Blackfan anemia. However, in patients with Duchenne muscular dystrophy, stem cell therapy still remains experimental. CASE PRESENTATION: We report the case of a nine-year-old boy of north Indian descent with Diamond-Blackfan anemia and Duchenne muscular dystrophy who underwent successful allogeneic hematopoietic stem cell transplantation. He is transfusion independent, and his Duchenne muscular dystrophy has shown no clinical deterioration over the past 45 months. His creatine phosphokinase levels have significantly decreased to 300 U/L from 14,000 U/L pre-transplant. The patient is 100% donor chimera in the hematopoietic system, and his muscle tissue has shown 8% to 10.4% cells of donor origin. CONCLUSION: Our patient's Diamond-Blackfan anemia was cured by allogeneic hematopoietic stem cell transplantation. The interesting clinical observation of a possible benefit in Duchenne muscular dystrophy cannot be ruled out. However, further clinical follow-up with serial muscle biopsies and molecular studies are needed to establish this finding. PMID- 21639929 TI - A baculovirus dual expression system-based vaccine confers complete protection against lethal challenge with H9N2 avian influenza virus in mice. AB - BACKGROUND: Avian influenza viruses of H9N2 subtype have become highly prevalent in avian species. Although these viruses generally cause only mild to moderate disease, they can infect a wide variety of species, including chickens, quail, turkeys, ducks, geese, pheasant, partridge, and pigeon, even transmitted to mammalian species, including humans, accelerating the efforts to devise protective strategies against them. RESULTS: The results showed that stronger immune responses were induced in a mouse model immunized with BV-Dual-HA than in those vaccinated with a DNA vaccine encoding the same antigen. Moreover, complete protection against lethal challenge with H9N2 virus was observed in mice. CONCLUSION: BV-Dual-HA could be utilized as a vaccine candidate against H9N2 virus infection. PMID- 21639931 TI - Complex evolutionary history of the Mexican stoneroller Campostoma ornatum Girard, 1856 (Actinopterygii: Cyprinidae). AB - BACKGROUND: Studies of the phylogeography of Mexican species are steadily revealing genetic patterns shared by different species, which will help to unravel the complex biogeographic history of the region. Campostoma ornatum is a freshwater fish endemic to montane and semiarid regions in northwest Mexico and southern Arizona. Its wide range of distribution and the previously observed morphological differentiation between populations in different watersheds make this species a useful model to investigate the biogeographic role of the Sierra Madre Occidental and to disentangle the actions of Pliocene tecto-volcanic processes vs Quaternary climatic change. Our phylogeographic study was based on DNA sequences from one mitochondrial gene (cytb, 1110 bp, n=285) and two nuclear gene regions (S7 and RAG1, 1822 bp in total, n=56 and 43, respectively) obtained from 18 to 29 localities, in addition to a morphological survey covering the entire distribution area. Such a dataset allowed us to assess whether any of the populations/lineages sampled deserve to be categorised as an evolutionarily significant unit. RESULTS: We found two morphologically and genetically well differentiated groups within C. ornatum. One is located in the northern river drainages (Yaqui, Mayo, Fuerte, Sonora, Casas Grandes, Santa Clara and Conchos) and another one is found in the southern drainages (Nazas, Aguanaval and Piaxtla). The split between these two lineages took place about 3.9 Mya (CI=2.1 5.9). Within the northern lineage, there was strong and significant inter-basin genetic differentiation and also several secondary dispersal episodes whit gene homogenization between drainages. Interestingly, three divergent mitochondrial lineages were found in sympatry in two northern localities from the Yaqui river basin. CONCLUSIONS: Our results indicate that there was isolation between the northern and southern phylogroups since the Pliocene, which was related to the formation of the ancient Nazas River paleosystem, where the southern group originated. Within groups, a complex reticulate biogeographic history for C. ornatum populations emerges, following the taxon pulse theory and mainly related with Pliocene tecto-volcanic processes. In the northern group, several events of vicariance promoted by river or drainage isolation episodes were found, but within both groups, the phylogeographic patterns suggest the occurrence of several events of river capture and fauna interchange. The Yaqui River supports the most diverse populations of C. ornatum, with several events of dispersal and isolation within the basin. Based on our genetic results, we defined three ESUs within C. ornatum as a first attempt to promote the conservation of the evolutionary processes determining the genetic diversity of this species. They will likely be revealed as a valuable tool for freshwater conservation policies in northwest Mexico, where many environmental problems concerning the use of water have rapidly arisen in recent decades. PMID- 21639930 TI - Use of imaging biomarkers to assess perfusion and glucose metabolism in the skeletal muscle of dystrophic mice. AB - BACKGROUND: Duchenne muscular dystrophy (DMD) is a severe neuromuscular disease that affects 1 in 3500 boys. The disease is characterized by progressive muscle degeneration that results from mutations in or loss of the cytoskeletal protein, dystrophin, from the glycoprotein membrane complex, thus increasing the susceptibility of contractile muscle to injury. To date, disease progression is typically assessed using invasive techniques such as muscle biopsies, and while there are recent reports of the use of magnetic resonance, ultrasound and optical imaging technologies to address the issue of disease progression and monitoring therapeutic intervention in dystrophic mice, our study aims to validate the use of imaging biomarkers (muscle perfusion and metabolism) in a longitudinal assessment of skeletal muscle degeneration/regeneration in two murine models of muscular dystrophy. METHODS: Wild-type (w.t.) and dystrophic mice (weakly affected mdx mice that are characterized by a point mutation in dystrophin; severely-affected mdx:utrn-/- (udx) mice that lack functional dystrophin and are null for utrophin) were exercised three times a week for 30 minutes. To follow the progression of DMD, accumulation of 18F-FDG, a measure of glucose metabolism, in both wild-type and affected mice was measured with a small animal PET scanner (GE eXplore Vista). To assess changes in blood flow and blood volume in the hind limb skeletal muscle, mice were injected intravenously with a CT contrast agent, and imaged with a small animal CT scanner (GE eXplore Ultra). RESULTS: In hind limb skeletal muscle of both weakly-affected mdx mice and in severely-affected udx mice, we demonstrate an early, transient increase in both 18F-FDG uptake, and in blood flow and blood volume. Histological analysis of H&E-stained tissue collected from parallel littermates demonstrates the presence of both inflammatory infiltrate and centrally-located nuclei, a classic hallmark of myofibrillar regeneration. In both groups of affected mice, the early transient response was succeeded by a progressive decline in muscle perfusion and metabolism; this was also evidenced histologically. CONCLUSIONS: The present study demonstrates the utility of non-invasive imaging biomarkers in characterizing muscle degeneration/regeneration in murine models of DMD. These techniques may now provide a promising alternative for assessing both disease progression and the efficacy of new therapeutic treatments in patients. PMID- 21639933 TI - Geometric nonlinear diffusion filter and its application to X-ray imaging. AB - BACKGROUND: Denoising with edge preservation is very important in digital x-ray imaging since it may allow us to reduce x-ray dose in human subjects without noticeable degradation of the image quality. In denoising filter design for x-ray imaging, edge preservation as well as noise reduction is of great concern not to lose detailed spatial information for accurate diagnosis. In addition to this, fast computation is also important since digital x-ray images are mostly comprised of large sized matrices. METHODS: We have developed a new denoising filter based on the nonlinear diffusion filter model. Rather than employing four directional gradients around the pixel of interest, we use geometric parameters derived from the local pixel intensity distribution in calculating the diffusion coefficients in the horizontal and vertical directions. We have tested the filter performance, including edge preservation and noise reduction, using low dose digital radiography and micro-CT images. RESULTS: The proposed denoising filter shows performance similar to those of nonlinear anisotropic diffusion filters (ADFs), one Perona-Malik ADF and the other Weickert's ADF in terms of edge preservation and noise reduction. However, the computation time has been greatly reduced. CONCLUSIONS: We expect the proposed denoising filter can be greatly used for fast noise reduction particularly in low-dose x-ray imaging. PMID- 21639932 TI - BEL/Pao retrotransposons in metazoan genomes. AB - BACKGROUND: Long terminal repeat (LTR) retrotransposons are a widespread kind of transposable element present in eukaryotic genomes. They are a major factor in genome evolution due to their ability to create large scale mutations and genome rearrangements. Compared to other transposable elements, little attention has been paid to elements belonging to the metazoan BEL/Pao subclass of LTR retrotransposons. No comprehensive characterization of these elements is available so far. The aim of this study was to describe all BEL/Pao elements in a set of 62 sequenced metazoan genomes, and to analyze their phylogenetic relationship. RESULTS: We identified a total of 7,861 BEL/Pao elements in 53 of our 62 study genomes. We identified BEL/Pao elements in 20 genomes where such elements had not been found so far. Our analysis shows that BEL/Pao elements are the second-most abundant class of LTR retrotransposons in the genomes we study, more abundant than Ty1/Copia elements, and second only to Ty3/Gypsy elements. They occur in multiple phyla, including basal metazoan phyla, suggesting that BEL/Pao elements arose early in animal evolution. We confirm findings from previous studies that BEL/Pao elements do not occur in mammals. The elements we found can be grouped into more than 1725 families, 1623 of which are new, previously unknown families. These families fall into seven superfamilies, only five of which have been characterized so far. One new superfamily is a major subdivision of the Pao superfamily which we propose to call Dan, because it is restricted to the genome of the zebrafish Danio rerio. The other new superfamily comprises 83 elements and is restricted to lower aquatic eumetazoans. We propose to call this superfamily Flow. BEL/Pao elements do not show any signs of recent horizontal gene transfer between distantly related species. CONCLUSIONS: In sum, our analysis identifies thousands of new BEL/Pao elements and provides new insights into their distribution, abundance, and evolution. PMID- 21639934 TI - Concurrent chemoradiotherapy with carboplatin followed by carboplatin and 5 fluorouracil in locally advanced nasopharyngeal carcinoma. AB - BACKGROUND: This study aimed to evaluate acute major toxicities, the response rate, 3-year overall survival and progression-free survival rate of locally advanced nasopharyngeal carcinoma patients on concurrent carboplatin chemoradiotherapy followed by carboplatin and 5-fluorouracil. METHODS: A prospective study of fifty patients diagnosed with locally advanced nasopharyngeal carcinoma received conventional radiation therapy with a total dose of 6600-7000 cGy in 6-7 weeks and concurrent chemotherapy of three cycles of carboplatin during radiotherapy, followed by adjuvant chemotherapy using carboplatin plus 5-fluorouracil for two cycles. RESULTS: Weight loss and mucositis were the two most common acute major grades 3-4 toxicities (42%). Myelosuppression occurred subsequently, including leukopenia (30%), neutropenia (20%), anemia (12%), and thrombocytopenia (6%). Only 8% of patients developed grades 3-4 nausea and vomiting. No patients had renal and electrolyte abnormalities. Regarding the response evaluation, 100% of patients achieved an objective response rate of the primary tumor (92% complete response, and 8% partial response). Similarly, all patients also achieved an objective response rate of the neck node (64% complete response and 36% partial response). The 3 year overall survival rate and progression-free survival rate were 89.7% and 72.7%, respectively. CONCLUSIONS: Concurrent chemoradiotherapy with carboplatin followed by carboplatin and 5- fluorouracil could be considered as an alternative regimen for locally advanced nasopharyngeal carcinoma patients pertaining to a good overall response rate, 3-year overall survival and progression-free survival rate with good tolerability. PMID- 21639935 TI - Is simulation training effective in increasing podiatrists' confidence in foot ulcer management? AB - BACKGROUND: Foot ulcers are a frequent reason for diabetes-related hospitalisation. Clinical training is known to have a beneficial impact on foot ulcer outcomes. Clinical training using simulation techniques has rarely been used in the management of diabetes-related foot complications or chronic wounds. Simulation can be defined as a device or environment that attempts to replicate the real world. The few non-web-based foot-related simulation courses have focused solely on training for a single skill or "part task" (for example, practicing ingrown toenail procedures on models). This pilot study aimed to primarily investigate the effect of a training program using multiple methods of simulation on participants' clinical confidence in the management of foot ulcers. METHODS: Sixteen podiatrists participated in a two-day Foot Ulcer Simulation Training (FUST) course. The course included pre-requisite web-based learning modules, practicing individual foot ulcer management part tasks (for example, debriding a model foot ulcer), and participating in replicated clinical consultation scenarios (for example, treating a standardised patient (actor) with a model foot ulcer). The primary outcome measure of the course was participants' pre- and post completion of confidence surveys, using a five-point Likert scale (1 = Unacceptable-5 = Proficient). Participants' knowledge, satisfaction and their perception of the relevance and fidelity (realism) of a range of course elements were also investigated. Parametric statistics were used to analyse the data. Pearson's r was used for correlation, ANOVA for testing the differences between groups, and a paired-sample t-test to determine the significance between pre- and post-workshop scores. A minimum significance level of p < 0.05 was used. RESULTS: An overall 42% improvement in clinical confidence was observed following completion of FUST (mean scores 3.10 compared to 4.40, p < 0.05). The lack of an overall significant change in knowledge scores reflected the participant populations' high baseline knowledge and pre-requisite completion of web-based modules. Satisfaction, relevance and fidelity of all course elements were rated highly. CONCLUSIONS: This pilot study suggests simulation training programs can improve participants' clinical confidence in the management of foot ulcers. The approach has the potential to enhance clinical training in diabetes-related foot complications and chronic wounds in general. PMID- 21639936 TI - Systemic epidermal nevus with involvement of the oral mucosa due to FGFR3 mutation. AB - BACKGROUND: Epidermal nevi (EN) represent benign congenital skin lesions following the lines of Blaschko. They result from genetic mosaicism, and activating FGFR3 and PIK3CA mutations have been implicated. CASE PRESENTATION: We report a female patient with a systemic keratinocytic nevus also involving the oral mucosa. Molecular genetic analysis revealed a mosaicism of the FGFR3 hotspot mutation R248C in the EN lesions of the skin and of the oral mucosa. The detection of the R248C mutation in a proportion of blood leukocytes and a slight scoliosis suggest an EN syndrome. CONCLUSIONS: Our results show that activating FGFR3 mutations can also affect the oral mucosa and that extracutaneous manifestations of EN syndrome can be subtle. We highlight the theoretical risk of the patient having an offspring with thanatophoric dysplasia as gonadal mosaicism for the R248C mutation cannot be excluded. PMID- 21639937 TI - Aberrant methylation of N-methyl-D-aspartate receptor type 2B (NMDAR2B) in non small cell carcinoma. AB - BACKGROUND: N-methyl-D-aspartate receptors (NMDAR) act as tumor suppressors of digestive malignancies. The expression and genetic methylation patterns of NMDAR2B in non-small cell lung cancer (NSCLC) are unknown. METHODS: The relationship between gene methylation and expression of NMDAR2B was analyzed in NSCLC cell lines (N = 9) and clinical tissues (N = 216). The cell lines were studied using RT-PCR and 5-aza-2'-deoxycytidine treatment, while the clinical tissues were examined by methylation specific real-time quantitative PCR and immunohistochemistry. Retrospective investigation of patient records was used to determine the clinical significance of NMDAR2B methylation. RESULTS: NMDAR2B was silenced in five of the nine cell lines; 5-aza-2'-deoxycytidine treatment restored expression, and was inversely correlated with methylation. Aberrant methylation of NMDAR2B, detected in 61% (131/216) of clinical NSCLC tissues, was inversely correlated with the status of protein expression in 20 randomly examined tumors. Aberrant methylation was not associated with clinical factors such as gender, age, histological type, or TNM stage. However, aberrant methylation was an independent prognostic factor in squamous cell carcinoma cases. CONCLUSIONS: Aberrant methylation of the NMDAR2B gene is a common event in NSCLC. The prognosis was significantly better for cases of squamous cell carcinoma in which NMDAR2B was methylated. It may have different roles in different histological types. PMID- 21639938 TI - Safety during the monitoring of diabetic patients: trial teaching course on health professionals and diabetics - SEGUDIAB study. AB - BACKGROUND: Safety for diabetic patients means providing the most suitable treatment for each type of diabetic in order to improve monitoring and to prevent the adverse effects of drugs and complications arising from the disease. The aim of this study is to analyze the effect of imparting educational interventions to health professionals regarding the safety of patients with Diabetes Mellitus (DM). METHODS: DESIGN: A cluster randomized trial with a control group. SETTING AND SAMPLE: The study analyzed ten primary healthcare centres (PHC) covering approximately 150,000 inhabitants. Two groups of 5 PHC were selected on the basis of their geographic location (urban, semi-urban and rural), their socio-economic status and the size of their PHC, The interventions and control groups were assigned at random. The study uses computerized patient records to individually assess subjects aged 45 to 75 diagnosed with type 1 and type 2 DM, who met the inclusion conditions and who had the variables of particular interest to the study. Trial: The educational interventions consisted of a standardized teaching course aimed at doctors and nurses. The course lasted 6 hours and was split into three 2-hour blocks with subsequent monthly refresher courses. MEASUREMENT: For the health professionals, the study used the Diabetes Attitude Scale (DAS-3) to assess their attitudes and motivation when monitoring diabetes. For the patients, the study assessed factors related to their degree of control over the disease at onset, 6, 12 and 24 months. MAIN VARIABLES: levels of HbA1c. ANALYSIS: The study analyzed the effect of the educational interventions both on the attitudes and motivations of health professionals and on the degree of control over the diabetes in both groups. DISCUSSION: Imparting educational interventions to health professionals would improve the monitoring of diabetic patients. The most effective model involves imparting the course to both doctors and nurses. However, these models have not been tested on our Spanish population within the framework of primary healthcare. TRIAL REGISTRATION: ClinicalTrials.gov: NCT01087541. PMID- 21639939 TI - Alpha-1 antitrypsin is a potential biomarker for hepatitis B. AB - BACKGROUND: Function exertion of specific proteins are key factors in disease progression, thus the systematical identification of those specific proteins is a prerequisite to understand various diseases. Though many proteins have been verified to impact on hepatitis, no systematical protein screening has been documented to hepatitis B virus (HBV) induced hepatitis, hindering the comprehensive understanding to this severe disease. AIM: To identify the major proteins in the progression of HBV infection from mild stage to severe stage. METHODS: We performed an integrated strategy by combining two-dimensional electrophoresis (2-DE), peptide mass fingerprinting (PMF) analysis, and tissue microarray techniques to screen the functional proteins and detect the localization of those proteins. RESULTS: Interestingly, MS/MS identification revealed the expression level of alpha-1 antitrypsin (AAT) was significantly elevated in serum samples from patients with severe chronic hepatitis. Immunoblotting with a specific AAT antibody confirmed that AAT is highly expressed in serum samples from patients with hepatic carcinoma and severe chronic hepatitis. Furthermore, we observed that AAT is with highest expression in normal tissue and cells, but lowest in hepatic carcinoma and severe chronic hepatitis tissues and cells, suggesting the specific secretion of AAT from tissues and cells to serum. CONCLUSION: These results suggest the possibility of AAT as a potential biomarker for hepatitis B in diagnosis. PMID- 21639940 TI - Integrating a family-focused approach into child obesity prevention: rationale and design for the My Parenting SOS study randomized control trial. AB - BACKGROUND: More than 20% of US children ages 2-5 yrs are classified as overweight or obese. Parents greatly influence the behaviors their children adopt, including those which impact weight (e.g., diet and physical activity). Unfortunately, parents often fail to recognize the risk for excess weight gain in young children, and may not be motivated to modify behavior. Research is needed to explore intervention strategies that engage families with young children and motivate parents to adopt behaviors that will foster healthy weight development. METHODS: This study tests the efficacy of the 35-week My Parenting SOS intervention. The intervention consists of 12 sessions: initial sessions focus on general parenting skills (stress management, effective parenting styles, child behavior management, coparenting, and time management) and later sessions apply these skills to promote healthier eating and physical activity habits. The primary outcome is change in child percent body fat. Secondary measures assess parent and child dietary intake (three 24-hr recalls) and physical activity (accelerometry), general parenting style and practices, nutrition- and activity related parenting practices, and parent motivation to adopt healthier practices. DISCUSSION: Testing of these new approaches contributes to our understanding of how general and weight-specific parenting practices influence child weight, and whether or not they can be changed to promote healthy weight trajectories. TRIAL REGISTRATION: ClinicalTrials.gov: NCT00998348. PMID- 21639942 TI - Persons with dementia missing in the community: is it wandering or something unique? AB - BACKGROUND: At some point in the disease process many persons with dementia (PWD) will have a missing incident and be unable to safely return to their care setting. In previous research studies, researchers have begun to question whether this phenomenon should continue to be called wandering since the antecedents and characteristics of a missing incident are dissimilar to accepted definitions of wandering in dementia. The purpose of this study was to confirm previous findings regarding the antecedents and characteristics of missing incidents, understand the differences between those found dead and alive, and compare the characteristics of a missing incident to that of wandering. METHODS: A retrospective design was used to analyse 325 newspaper reports of PWD missing in the community. RESULTS: The primary antecedent to a missing incident, particularly in community-dwelling PWD, was becoming lost while conducting a normal and permitted activity alone in the community. The other common antecedent was a lapse in supervision with the expectation that the PWD would remain in a safe location but did not. Deaths most commonly occurred in unpopulated areas due to exposure and drowning. Those who died were found closer to the place last seen and took longer to find, but there were no significant differences in gender or age. The key characteristics of a missing incident were: unpredictable, non repetitive, temporally appropriate but spatially-disordered, and while using multiple means of movement (walking, car, public transportation). Missing incidents occurred without the discernible pattern present in wandering such as lapping or pacing, repetitive and temporally-disordered. CONCLUSIONS: This research supports the mounting evidence that the concept of wandering, in its formal sense, and missing incidents are two distinct concepts. It will be important to further develop the concept of missing incidents by identifying the differences and similarities from wandering. This will allow a more targeted assessment and intervention strategy for each problem. PMID- 21639941 TI - Acupuncture for the treatment of severe acute pain in herpes zoster: results of a nested, open-label, randomized trial in the VZV Pain Study. AB - BACKGROUND: Data on the potential efficacy of acupuncture (AC) in controlling intense or very intense pain in patients with Herpes Zoster (HZ) has not been so far adequately assessed in comparison with standard pharmacological treatment (ST) by a controlled trial design. METHODS: Within the VZV Pescara study, pain was assessed in HZ patients on a Visual Analogue Scale (VAS) and by the McGill Pain Questionnaire (MPQ) both at the beginning and at the end of treatment. Response rates, mean changes in pain intensity, differences in total pain burden with an area-under-the-curve (AUC) method over a 1-year follow-up and differences in the incidence of Post-Herpetic Neuralgia (PHN) were evaluated. RESULTS: One hundred and two patients were randomized to receive either AC (n = 52) or ST (n = 50) for 4 weeks. Groups were comparable regarding age, sex, pain intensity at presentation and missed antiviral prescription. Both interventions were largely effective. No significant differences were observed in response rates (81.6% vs 89.2%, p = 0.8), mean reduction of VAS (4.1 +/- 2.3 vs 4.9 +/- 1.9, p = 0.12) and MPQ scores (1.3 +/- 0.9 vs 1.3 +/- 0.9, p = 0.9), incidence of PHN after 3 months (48.4% vs 46.8%, p = 0.5), and mean AUC during follow-up (199 +/- 136 vs 173 +/- 141, p = 0.4). No serious treatment-related adverse event was observed in both groups. CONCLUSIONS: This controlled and randomized trial provides the first evidence of a potential role of AC for the treatment of acute herpetic pain. TRIAL REGISTRATION: ChiCTR-TRC-10001146. PMID- 21639943 TI - Trends in chlamydia and gonorrhea positivity among heterosexual men and men who have sex with men attending a large urban sexual health service in Australia, 2002-2009. AB - BACKGROUND: To determine whether chlamydia positivity among heterosexual men (MSW) and chlamydia and gonorrhea positivity among men who have sex with men (MSM), are changing. METHODS: Computerized records for men attending a large sexual health clinic between 2002 and 2009 were analyzed. Chlamydia and gonorrhea positivity were calculated and logistic regression used to assess changes over time. RESULTS: 17769 MSW and 8328 MSM tested for chlamydia and 7133 MSM tested for gonorrhea. In MSW, 7.37% (95% CI: 6.99-7.77) were chlamydia positive; the odds of chlamydia positivity increased by 4% per year (OR = 1.04; 95% CI: 1.01 1.07; p = 0.02) after main risk factors were adjusted for. In MSM, 3.70% (95% CI: 3.30-4.14) were urethral chlamydia positive and 5.36% (95% CI: 4.82-5.96) were anal chlamydia positive; positivity could not be shown to have changed over time. In MSM, 3.05% (95% CI: 2.63-3.53) tested anal gonorrhea positive and 1.83% (95% CI: 1.53-2.18) tested pharyngeal gonorrhea positive. Univariate analysis found the odds of anal gonorrhea positivity had decreased (OR = 0.93; 95% CI: 0.87 1.00; p = 0.05), but adjusting for main risk factors resulted in no change. Urethral gonorrhea cases in MSM as a percentage of all MSM tested for gonorrhea also fell (p < 0.001). CONCLUSIONS: These data suggest that chlamydia prevalence in MSW is rising and chlamydia and gonorrhea prevalence among MSM is stable or declining. High STI testing rates among MSM in Australia may explain differences in STI trends between MSM and MSW. PMID- 21639944 TI - A chalcone-related small molecule that induces methuosis, a novel form of non apoptotic cell death, in glioblastoma cells. AB - BACKGROUND: Methuosis is a unique form of non-apoptotic cell death triggered by alterations in the trafficking of clathrin-independent endosomes, ultimately leading to extreme vacuolization and rupture of the cell. RESULTS: Here we describe a novel chalcone-like molecule, 3-(2-methyl-1H- indol-3-yl)-1-(4 pyridinyl)-2-propen-1-one (MIPP) that induces cell death with the hallmarks of methuosis. MIPP causes rapid accumulation of vacuoles derived from macropinosomes, based on time-lapse microscopy and labeling with extracellular fluid phase tracers. Vacuolization can be blocked by the cholesterol-interacting compound, filipin, consistent with the origin of the vacuoles from non-clathrin endocytic compartments. Although the vacuoles rapidly acquire some characteristics of late endosomes (Rab7, LAMP1), they remain distinct from lysosomal and autophagosomal compartments, suggestive of a block at the late endosome/lysosome boundary. MIPP appears to target steps in the endosomal trafficking pathway involving Rab5 and Rab7, as evidenced by changes in the activation states of these GTPases. These effects are specific, as other GTPases (Rac1, Arf6) are unaffected by the compound. Cells treated with MIPP lose viability within 2-3 days, but their nuclei show no evidence of apoptotic changes. Inhibition of caspase activity does not protect the cells, consistent with a non-apoptotic death mechanism. U251 glioblastoma cells selected for temozolomide resistance showed sensitivity to MIPP-induced methuosis that was comparable to the parental cell line. CONCLUSIONS: MIPP might serve as a prototype for new drugs that could be used to induce non-apoptotic death in cancers that have become refractory to agents that work through DNA damage and apoptotic mechanisms. PMID- 21639945 TI - Evaluating methods for ranking differentially expressed genes applied to microArray quality control data. AB - BACKGROUND: Statistical methods for ranking differentially expressed genes (DEGs) from gene expression data should be evaluated with regard to high sensitivity, specificity, and reproducibility. In our previous studies, we evaluated eight gene ranking methods applied to only Affymetrix GeneChip data. A more general evaluation that also includes other microarray platforms, such as the Agilent or Illumina systems, is desirable for determining which methods are suitable for each platform and which method has better inter-platform reproducibility. RESULTS: We compared the eight gene ranking methods using the MicroArray Quality Control (MAQC) datasets produced by five manufacturers: Affymetrix, Applied Biosystems, Agilent, GE Healthcare, and Illumina. The area under the curve (AUC) was used as a measure for both sensitivity and specificity. Although the highest AUC values can vary with the definition of "true" DEGs, the best methods were, in most cases, either the weighted average difference (WAD), rank products (RP), or intensity-based moderated t statistic (ibmT). The percentages of overlapping genes (POGs) across different test sites were mainly evaluated as a measure for both intra- and inter-platform reproducibility. The POG values for WAD were the highest overall, irrespective of the choice of microarray platform. The high intra- and inter-platform reproducibility of WAD was also observed at a higher biological function level. CONCLUSION: These results for the five microarray platforms were consistent with our previous ones based on 36 real experimental datasets measured using the Affymetrix platform. Thus, recommendations made using the MAQC benchmark data might be universally applicable. PMID- 21639946 TI - Genetic factors associated with patient-specific warfarin dose in ethnic Indonesians. AB - BACKGROUND: CYP2C9 and VKORC1 are two major genetic factors associated with inter individual variability in warfarin dose. Additionally, genes in the warfarin metabolism pathway have also been associated with dose variance. We analyzed Single Nucleotide Polymorphisms (SNPs) in these genes to identify genetic factors that might confer warfarin sensitivity in Indonesian patients. METHODS: Direct sequencing method was used to identify SNPs in CYP2C9, VKORC1, CYP4F2, EPHX1, PROC and GGCX genes in warfarin-treated patients. Multiple linear regressions were performed to model the relationship warfarin daily dose requirement with genetic and non-genetic variables measured and used to develop a novel algorithm for warfarin dosing. RESULTS: From the 40 SNPs analyzed, CYP2C9 rs17847036 and VKORC1 rs9923231 showed significant association with warfarin sensitivity. In our study population, no significant correlation could be detected between CYP2C9*3, CYP2C9C-65 (rs9332127), CYP4F2 rs2108622, GGCX rs12714145, EPHX1 rs4653436 and PROC rs1799809 with warfarin sensitivity. CONCLUSIONS: VKORC1 rs9923231 AA and CYP2C9 rs17847036 GG genotypes were associated with low dosage requirements of most patients (2.05 +/- 0.77 mg/day and 2.09 +/- 0.70 mg/day, respectively). CYP2C9 and VKORC1 genetic variants as well as non-genetic factors such as age, body weight and body height account for 15.4% of variance in warfarin dose among our study population. Additional analysis of this combination could allow for personalized warfarin treatment in ethnic Indonesians. PMID- 21639947 TI - Clinical, electroretinographic and histomorphometric evaluation of the retina in sheep with natural scrapie. AB - BACKGROUND: The retina is part of the diencephalon in a peripheral location and may be involved in prion diseases. Retinal function and structural changes were assessed in naturally scrapie-affected red face Manech ewes presenting the classical signs of the disease, and clinically healthy age-matched subjects for controls. Ophthalmic examination was done prior to electroretinography (ERG), which was carried out under conditions that allowed photopic and scotopic activities to be assessed. Histomorphometry of the inner and outer retinal layers was performed post-mortem, and retinas were also examined for evidence of abnormal prion protein (PrPSc) accumulation and glial fibrillary acidic protein (GFAP) upregulation as a marker of gliosis. Scrapie status was determined by examination of brain tissue RESULTS: Ocular reflexes and ophthalmoscopy did not reveal any difference between scrapie affected and control animals. Although the light-and dark-adapted ERG responses of both rod-and cone-mediated functions had a similar waveform in scrapie-affected and control sheep, a significant reduction in the amplitude of the ERG a-and b-waves was observed in affected animals compared to controls. These functional alterations were correlated with a substantial loss of cells in the outer nuclear layer (ONL), lengthening and disorganization in photoreceptor segments, and substantial reduction in cellularity and thickness of the inner nuclear layer (INL). The degenerative changes in the INL and ONL were most marked in the central and paracentral areas of the scrapie retinas, and were accompanied in all scrapie retinas by PrPSc deposition in the ganglion cell and synaptic layers. GFAP immunoreactivity was mainly increased in the ganglion cell and inner plexiform layers. CONCLUSIONS: No appreciable fundoscopic changes were observed in the scrapie-affected ewes although reproducible changes in retinal function as measured by ERG were observed in these animals. The alterations in the receptoral and post-receptoral pathways corresponded to the degenerative lesions observed in the ONL and INL of the scrapie retinas. The retinal degeneration was associated with prion protein infectivity which presumably spread via the optic nerve. PMID- 21639949 TI - Endocrine resistance in breast cancer: new roles for ErbB3 and ErbB4. AB - Endocrine resistance is a major limitation to the successful treatment of estrogen receptor-positive (ER(+)) breast cancer, and the EGFR (epidermal growth factor receptor) and ErbB-2 receptor tyrosine kinases are involved in this process. A recent study now implicates the other two ErbB family members, ErbB-3 and -4. Exposure of ER+ breast cancer cells to the pure antiestrogen, fulvestrant, increased levels of ErbB-3 or ErbB-4 and sensitivity to the growth stimulatory effects of heregulin beta1, a potent ligand for these receptors. Thus, the initial growth-inhibitory effects of fulvestrant appear compromised by cellular plasticity that allows rapid compensatory growth stimulation via ErbB 3/4. Further evaluation of pan-ErbB receptor inhibitors in endocrine-resistant disease appears warranted. PMID- 21639948 TI - The lamin protein family. AB - The lamins are the major architectural proteins of the animal cell nucleus. Lamins line the inside of the nuclear membrane, where they provide a platform for the binding of proteins and chromatin and confer mechanical stability. They have been implicated in a wide range of nuclear functions, including higher-order genome organization, chromatin regulation, transcription, DNA replication and DNA repair. The lamins are members of the intermediate filament (IF) family of proteins, which constitute a major component of the cytoskeleton. Lamins are the only nuclear IFs and are the ancestral founders of the IF protein superfamily. Lamins polymerize into fibers forming a complex protein meshwork in vivo and, like all IF proteins, have a tripartite structure with two globular head and tail domains flanking a central alpha-helical rod domain, which supports the formation of higher-order polymers. Mutations in lamins cause a large number of diverse human diseases, collectively known as the laminopathies, underscoring their functional importance. PMID- 21639950 TI - The new era of autoimmune disease research. AB - Recent genome-wide association studies have advanced our understanding of genetic factors that underlie systemic lupus erythematosus (SLE), a multifactorial autoimmune disease characterized by various clinical manifestations. SLE also has an environmental component, which can trigger or exacerbate the disease. Despite extensive efforts aimed at elucidating the cellular and biological abnormalities that arise in the immune system of patients with SLE, its pathology remains unclear. Lee and colleagues recently carried out gene expression profiling of patients with SLE followed by bioinformatics analysis and discovered the existence of abnormal regulatory networks and potential key molecules. The authors found that ATP synthesis and DNA repair pathways may be involved in the pathogenesis, providing a potential explanation for photosensitivity experienced by patients with SLE. PMID- 21639951 TI - Negative regulation of NF-kappaB and its involvement in rheumatoid arthritis. AB - The transcription factor NF-kappaB plays crucial roles in the regulation of inflammation and immune responses, and inappropriate NF-kappaB activity has been linked with many autoimmune and inflammatory diseases, including rheumatoid arthritis. Cells employ a multilayered control system to keep NF-kappaB signalling in check, including a repertoire of negative feedback regulators ensuring termination of NF-kappaB responses. Here we will review various negative regulatory mechanisms that have evolved to control NF-kappaB signalling and which have been implicated in the pathogenesis of rheumatoid arthritis. PMID- 21639952 TI - The beta-secretase enzyme BACE1 as a therapeutic target for Alzheimer's disease. AB - Amyloid plaques are defining histopathologic lesions in the brains of Alzheimer's disease (AD) patients and are composed of the amyloid-beta peptide, which is widely considered to play a critical role in the pathogenesis of AD. The beta secretase, or beta-site amyloid precursor protein cleaving enzyme 1 (BACE1; also called Asp2, memapsin 2), is the enzyme that initiates the generation of amyloid beta. Consequently, BACE1 is an attractive drug target for lowering cerebral levels of amyloid beta for the treatment or prevention of AD. Much has been learned about BACE1 since its discovery over 10 years ago. In the present article, we review BACE1 properties and characteristics, cell biology, in vivo validation, substrates, therapeutic potential, and inhibitor drug development. Studies relating to the physiological functions of BACE1 and the promise of BACE1 inhibition for AD will also be discussed. We conclude that therapeutic inhibition of BACE1 should be efficacious for AD, although careful titration of the drug dose may be necessary to limit mechanism-based side effects. PMID- 21639953 TI - Bright days for yeast research. AB - A report on the British Yeast Group Meeting, Brighton, UK, 23-25 March 2011. PMID- 21639954 TI - Combining DNA-microarray data in systemic lupus erythematosus. AB - Systemic lupus erythematosus is a systemic, heterogeneous autoimmune disease. Understanding of its molecular complexity is incomplete and there is a need to identify new therapeutic targets and to optimize criteria for its diagnosis, assessment and prognosis. Recently, Arasappan and colleagues have described a new meta-analysis method that enables data analysis across different DNA-microarray datasets to identify genes and processes relevant to systemic lupus erythematosus. Their study provides a simple and valuable meta-analysis method for the selection of biomarkers and pathways in disease.See related research by Arasappan et al.: http://www.biomedcentral.com/1741-7015/9/65. PMID- 21639955 TI - Dedifferentiation rescues senescence of progeria cells but only while pluripotent. AB - Hutchinson-Gilford progeria syndrome (HGPS) is a genetic disease in which children develop pathologies associated with old age. HGPS is caused by a mutation in the LMNA gene, resulting in the formation of a dominant negative form of the intermediate filament, nuclear structural protein lamin A, termed progerin. Expression of progerin alters the nuclear architecture and heterochromatin, affecting cell cycle progression and genomic stability. Two groups recently reported the successful generation and characterization of induced pluripotent stem cells (iPSCs) from HGPS fibroblasts. Remarkably, progerin expression and senescence phenotypes are lost in iPSCs but not in differentiated progeny. These new HGPS iPSCs are valuable for characterizing the role of progerin in driving HGPS and aging and for screening therapeutic strategies to prevent or delay cell senescence. PMID- 21639956 TI - Spotlight on geminin. AB - In the previous issue of Breast Cancer Research, Gardner and co-workers describe a novel interaction between Geminin, a protein that prevents reinitiation of DNA replication, and Topoisomerase IIalpha (TopoIIalpha), an enzyme essential for removing catenated intertwines between sister chromatids. Geminin facilitates the action of TopoIIalpha, thereby promoting termination of DNA replication at the same time it inhibits initiation. In this manner, Geminin ensures that cells duplicate their genome once, but only once, each time they divide. Remarkably, either depletion of Geminin or over-expression of Geminin inhibits the action of TopoIIalpha, thereby making Geminin an excellent target for cancer chemotherapy. PMID- 21639957 TI - Soluble CD40 ligand, a mediator of sepsis or of transfusion-related adverse effects? PMID- 21639959 TI - Annual conference on hereditary cancers 2009 szczecin, poland. 10-11 december 2009. Abstracts. PMID- 21639958 TI - Peri-operative hemodynamic therapy: only large clinical trials can resolve our uncertainty. AB - In this issue of Critical Care, Cecconi and colleagues report the findings of a small trial of goal-directed hemodynamic therapy (GDT) in patients undergoing major orthopedic surgery under regional anesthesia. This is an interesting trial and the first of which we are aware to test the efficacy of GDT in this patient group. The findings suggest that this intervention is associated with improved adverse event rates after surgery. However, in this trial, as in all small trials of GDT, the potential for bias leaves some uncertainty regarding how widely the findings should be implemented. Such limitations may be impossible to completely eliminate from trials of complex interventions, but large multicenter trials may allow us to substantially decrease bias and improve the generalizability of the findings. PMID- 21639961 TI - Secretory endometrial adenocarcinoma. PMID- 21639960 TI - Copd. AB - INTRODUCTION: Chronic obstructive pulmonary disease (COPD) is a disease state characterised by airflow limitation that is not fully reversible. The airflow limitation is usually progressive and associated with an abnormal inflammatory response of the lungs to noxious particles or gases. Classically, it is thought to be a combination of emphysema and chronic bronchitis, although only one of these may be present in some people with COPD. The main risk factor for the development and deterioration of COPD is smoking. METHODS AND OUTCOMES: We conducted a systematic review and aimed to answer the following clinical questions: What are the effects of maintenance drug treatment in stable COPD? What are the effects of smoking cessation interventions in people with stable COPD? What are the effects of non-drug interventions in people with stable COPD? We searched: Medline, Embase, The Cochrane Library, and other important databases up to April 2010 (Clinical Evidence reviews are updated periodically; please check our website for the most up-to-date version of this review). We included harms alerts from relevant organisations such as the US Food and Drug Administration (FDA) and the UK Medicines and Healthcare products Regulatory Agency (MHRA). RESULTS: We found 119 systematic reviews, RCTs, or observational studies that met our inclusion criteria. We performed a GRADE evaluation of the quality of evidence for interventions. CONCLUSIONS: In this systematic review, we present information relating to the effectiveness and safety of the following interventions: alpha(1) antitrypsin, antibiotics (prophylactic), anticholinergics (inhaled), beta(2) agonists (inhaled), corticosteroids (oral and inhaled), general physical activity enhancement, inspiratory muscle training, nutritional supplementation, mucolytics, oxygen treatment (long-term domiciliary treatment), peripheral muscle strength training, psychosocial and pharmacological interventions for smoking cessation, pulmonary rehabilitation, and theophylline. PMID- 21639962 TI - Attitudes among Toronto obstetricians towards vaginal breech delivery. AB - OBJECTIVE: The recent SOGC guidelines allow for selective vaginal delivery of breech presentations, following an eight-year period during which vaginal breech delivery was discouraged based on the results of the Term Breech Trial (TBT). We sought to determine the effect of publication of this guideline on the acceptance of vaginal breech delivery by obstetricians and to correlate obstetricians' attitudes with actual practice. METHODS: A survey was sent to all obstetricians practising in five teaching hospitals in Toronto exploring their attitudes towards, and comfort with, vaginal breech delivery in various clinical situations. We correlated these with their graduation year in relation to the publication of the TBT. We also reviewed the obstetrical database of the largest teaching hospital in Toronto to see if these attitudes correlated with actual practice. RESULTS: The vaginal breech delivery rate, which was declining prior to publication of the TBT, plummeted after it. Our survey found that most practitioners (50% to 80%) would be willing to provide vaginal breech delivery in defined conditions, with more experienced obstetricians being more comfortable with offering vaginal breech delivery. However, despite these attitudes, the vaginal breech delivery rate during the period surveyed was only 3% (6/195). CONCLUSIONS: In the eight years between publication of the TBT and the new guidelines, very few vaginal breech deliveries were performed. Our survey indicates that most obstetricians have accepted the new guidelines; however, it seems that actual practice is lagging behind. The recent SOGC guidelines seem to have changed attitudes, but without changes in training and practical support, it seems unlikely that the trend for very few vaginal breech deliveries to be performed will be reversed. PMID- 21639963 TI - Higher caesarean section rates in women with higher body mass index: are we managing labour differently? AB - BACKGROUND: Higher body mass index has been associated with an increased risk of Caesarean section. The effect of differences in labour management on this association has not yet been evaluated. METHODS: We conducted a cohort study using data from the McGill Obstetrics and Neonatal Database for deliveries taking place during a 10-year period. Women's BMI at delivery was categorized as normal (20 to 24.9), overweight (25 to 29.9), obese (30 to 39.9), or morbidly obese (>= 40). We evaluated the effect of the management of labour on the need for Caesarean section using unconditional logistic regression models. RESULTS: Data were available for 11 922 women, of whom 2289 women had normal weight, 5663 were overweight, 3730 were obese, and 240 were morbidly obese. After adjustment for known confounding variables, increased BMI category was associated with an overall increase in the use of oxytocin and in the use of epidural analgesia, and with a decrease in use of forceps and vacuum extraction among second stage deliveries. Higher BMI was also found to be associated with earlier decisions to perform a Caesarean section in the second stage of labour. When adjusted for these differences in the management of labour, the increasing rate of Caesarean section observed with increasing BMI category was markedly attenuated (P < 0.001). CONCLUSION: Women with an increased BMI are managed differently in labour than women of normal weight. This difference in management in part explains the increased rate of Caesarean section observed with higher BMI. PMID- 21639964 TI - Developmental origin of chorionic villus cultures from spontaneous abortion and chorionic villus sampling. AB - OBJECTIVE: Chorionic villus cultures from spontaneous abortions and chorionic villus sampling (CVS) are routinely used for clinical cytogenetic analysis. Although these cultures are assumed to represent the chorionic villus mesenchymal core, and therefore the inner cell mass (ICM) of the blastocyst, immunochemical studies using a true trophoblast-specific marker to definitively rule out trophoblast contamination have not been done. Therefore, we used cytokeratin-7 (CK7), a trophoblast-specific marker, to assess the developmental origin of these chorionic villus cultures. METHODS: We assessed chorionic villus cultures from CVS and spontaneous abortions for CK7 immunostaining (n = 20). RESULTS: Cultures from both CVS and spontaneous abortions showed little or no CK7 staining (<= 1%). CONCLUSION: Chorionic villus cultures from CVS and spontaneous abortions exhibit little or no trophoblast contamination. They are therefore representative of the villus mesenchymal core and ultimately originate from the ICM of the blastocyst. PMID- 21639965 TI - Pregnancy and neonatal outcomes of women with reactive syphilis serology in Alberta, 2002 to 2006. AB - OBJECTIVES: To describe the maternal characteristics, diagnosis, and pregnancy, and the neonatal outcomes of pregnant women with reactive syphilis serology in a Canadian cohort. METHODS: We conducted a retrospective chart review of pregnant women in Alberta with reactive syphilis serology between 2002 and 2006. Clinical staging of syphilis in mothers and infants was determined using provincial and national surveillance criteria. RESULTS: Seventy-five pregnancies met the inclusion criteria. Thirty women were adequately treated pre-conception, 20 women had infectious syphilis (10 primary, 5 secondary, 5 early latent), 24 had late latent syphilis, and one had disease of unknown stage. Seven infants with congenital syphilis and one infant with presumed congenital syphilis were born to women with primary (n = 4), secondary (n = 2), early latent (n = 1), and unknown stage (n = 1) syphilis. Treatment was provided prior to delivery in one woman; five women did not access prenatal care. Four infants had long-term sequelae. CONCLUSION: All infants with congenital syphilis were born to women with infectious syphilis who had limited prenatal care. Initiatives to reach women at high risk are required to decrease the incidence of congenital syphilis. PMID- 21639966 TI - Pregnant women's perception of risk with use of the H1N1 vaccine. AB - OBJECTIVE: During the H1N1 influenza pandemic in 2009, The Motherisk Program, a counselling service providing teratology information, received many calls from pregnant women inquiring about the safety of the H1N1 vaccine. We wished to explore pregnant women's perception of risk and the factors associated with deciding whether or not to receive the vaccine. METHODS: Pregnant women who called Motherisk between October 1 and November 30, 2009, requesting counselling regarding the safety of the H1N1 vaccine, and who had not yet received the vaccine, were contacted for follow-up using a structured questionnaire. RESULTS: One hundred thirty women completed the questionnaire; 104 (80%) had received the H1N1 vaccination following their call to Motherisk, and 26 (20%) had not. More than 70% of the women cited confusing and frightening information in the media as a trigger for their concern, prompting them to call Motherisk. Sixty percent stated that information from their primary health care providers or Motherisk contributed to their decision making. CONCLUSION: The H1N1 vaccination rate in pregnant women who contacted Motherisk was higher than the rate in the general population, as many followed Motherisk's recommendation to receive the vaccine. During this period, the media appeared to provide pregnant women with confusing information. In any future pandemic scare, accessibility to primary health care providers or specialized information services such as Motherisk will be key to providing guidance for pregnant women. PMID- 21639967 TI - Choosing the right patient: planning for laparotomy or laparoscopy in the patient with endometrial cancer. AB - OBJECTIVE: Endometrial cancer remains the most commonly diagnosed gynaecologic cancer in North America. The staging and initial treatment of endometrial cancer involves surgery. Laparoscopic surgery is increasingly used as an alternative to laparotomy. Patient selection for laparoscopy can be optimized by examining factors involved in both the choice of surgical approach and the ultimate procedure performed. We wished to identify factors that might be barriers to laparoscopic surgery in women with endometrial cancer who had surgery performed by the gynaecologic oncology group at the Capital District Health Authority (CDHA) in Halifax, Nova Scotia. METHODS: We conducted a retrospective review of the records of women with endometrial cancer, histologically confirmed preoperatively between 2005 and 2007, who underwent surgery at the CDHA. RESULTS: Between 2005 and 2007 in Nova Scotia, 428 cases of endometrial cancer were diagnosed, and 289 women with a preoperative diagnosis of endometrial cancer underwent surgery at the CDHA. Of these, 66.1% (191/289) underwent a planned laparotomy, and 33.9% (98/289) had a planned laparoscopy. The proportion of attempted laparoscopies increased from 21.9% to 57.1% (P = 0.002) over time, while there was no change in the conversion rate (P = 0.23). Patients with abnormal findings on pelvic examination were more likely to have a laparotomy (RR = 1.5; 95% CI 1.34 to 1.68). Independent predictors of laparoscopic conversion to laparotomy were age 75 years or over (P = 0.03) and non-endometrioid histology (P = 0.002). CONCLUSION: Our data identify age and non-endometrioid histology as independent factors for conversion of surgery for endometrial cancer from laparoscopy to laparotomy. With this information we can optimize patient selection for laparoscopic surgery. Patients undergoing a conversion to laparotomy do not have a significant increase in surgery time or perioperative morbidity. PMID- 21639968 TI - Sonographic findings and clinical outcomes in women with massive subchorionic hematoma detected in the second trimester. AB - BACKGROUND: Small hematomas on the placental surface, termed subamniotic hemorrhage, are a common finding either at the routine 18- to 20-week anatomy ultrasound or at subsequent assessments of fetal growth and well-being. Hemorrhage beneath or at the edge of the placenta, or behind an isolated area of the fetal membranes, is of greater concern. THE CASES: We describe the ultrasound findings and clinical outcomes in two women with a diagnosis of massive intrauterine hematoma arising from the fetal membranes. Both required blood transfusion because of the extent of concealed and revealed bleeding. In each case the initial placental appearances and uterine artery Doppler studies were normal. Both hematomas resolved with growth of the fetus and amniotic sac. Each neonate survived the perinatal period favourably. One was born vaginally at 32 weeks' gestation following premature preterm rupture of the membranes, and the second was born by emergency Caesarean section at 37 weeks because of a recurrence of antepartum hemorrhage. CONCLUSION: Large intrauterine hematomas may be acutely detrimental to maternal health in the second trimester. Ultrasound assessment of the placenta is useful to define the perinatal prognosis and may demonstrate gradual resolution. Despite a dramatic initial presentation, this finding may be compatible with a favourable outcome. PMID- 21639970 TI - Implementing an altruistic sperm donation program in Canada. PMID- 21639969 TI - Prevention of adverse pregnancy outcomes with low-dose ASA in early pregnancy: new perspectives for future randomized trials. AB - Recent evidence suggests that treatment with low-dose acetylsalicylic acid (ASA) started early in pregnancy could prevent preeclampsia and intrauterine growth restriction (IUGR), two complications involving placental dysfunction. Preterm birth could also potentially be prevented, suggesting that it could share mechanisms of disease with preeclampsia and intrauterine growth restriction. Because there is new evidence that placental dysfunction can be predicted as early as in the first trimester, we argue that there is a need for randomized controlled trials of low-dose ASA for the prevention of preeclampsia, IUGR, and possibly preterm birth among nulliparous women with early indicators of placental dysfunction. PMID- 21639971 TI - SOGC Clinical Practice Guideline. Ultrasonographic cervical length assessment in predicting preterm birth in singleton pregnancies. AB - OBJECTIVES: To review (1) the use of ultrasonographic-derived cervical length measurement in predicting preterm birth and (2) interventions associated with a short cervical length. OUTCOMES: Reduction in rates of prematurity and/or better identification of those at risk, as well as possible prevention of unnecessary interventions. EVIDENCE: Published literature was retrieved through searches of PubMed and The Cochrane Library up to December 2009, using appropriate controlled vocabulary and key words (preterm labour, ultrasound, cervix, incompetent cervix, transvaginal, transperineal, cervical length, fibronectin). Results were restricted to general and systematic reviews, randomized controlled trials/controlled clinical trials, and observational studies. There were no date or language restrictions. Grey (unpublished) literature was identified through searching the websites of health technology assessment and health technology assessment-related agencies, clinical practice guideline collections, clinical trial registries, and national and international medical specialty societies. VALUES: The evidence and this guideline were reviewed by the Diagnostic Imaging Committee and the Maternal Fetal Medicine Committee of the Society of Obstetricians and Gynaecologists of Canada, and the recommendations were made according to the guidelines developed by The Canadian Task Force on Preventive Health Care (Table 1). BENEFITS, HARMS, AND COSTS: Preterm birth is a leading cause of perinatal morbidity and mortality. Use of the ultrasonographic technique reviewed in this guideline may help identify women at risk of preterm birth and, in some circumstances, lead to interventions that may reduce the rate of preterm birth. SPONSORS: The Society of Obstetricians and Gynaecologists of Canada. PMID- 21639974 TI - Synthesis and Characterization of Ciprofloxacin Pendant Antibacterial Cationic Polymers. AB - A methacrylate monomer containing ciprofloxacin (CPF) was synthesized, followed by the free-radical co-polymerization of the methacrylate monomer, acrylamide and diallyl dimethyl ammonium chloride (DADMAC). The resulting antimicrobial cationic tripolymers were characterized by (1)H-NMR, gel-permeation chromatography, thermogravimetric analysis and differential scanning calorimetry. The in vitro antibacterial activity of the monomer, as well as of the polymers, was investigated against E. coli. The CPF pendant tripolymer exhibited excellent antibacterial activities, thus permitting the cationic tripolymers to be used as antibacterial intensifier, retention or filtration aid for various hygiene products, including those based on cellulose fibres. PMID- 21639972 TI - SOGC Clinical Practice Guideline. Magnesium sulphate for fetal neuroprotection. AB - OBJECTIVE: To provide guidelines for the use of antenatal magnesium sulphate (MgSO4) for fetal neuroprotection of the preterm infant. OPTIONS: Antenatal MgSO4 administration should be considered for fetal neuroprotection when women present at <= 31+6 weeks with imminent preterm birth, defined as a high likelihood of birth because of active labour with cervical dilatation >= 4 cm, with or without preterm pre-labour rupture of membranes, and/or planned preterm birth for fetal or maternal indications. There are no other known fetal neuroprotective agents. OUTCOMES: The outcomes measured are the incidence of cerebral palsy (CP) and neonatal death. EVIDENCE: Published literature was retrieved through searches of PubMed or Medline, CINAHL, and the Cochrane Library in May 2010, using appropriate controlled vocabulary and key words (magnesium sulphate, cerebral palsy, preterm birth). Results were restricted to systematic reviews, randomized controlled trials, and relevant observational studies. There were no date or language restrictions. Searches were updated on a regular basis and incorporated in the guideline to August 2010. Grey (unpublished) literature was identified through searching the websites of health technology assessment and health technology assessment-related agencies, clinical practice guideline collections, clinical trial registries, and national and international medical specialty societies. VALUES: The quality of evidence was rated using the criteria described in the Report of the Canadian Task Force on Preventive Health Care (Table 1). BENEFITS, HARMS, AND COSTS: Antenatal magnesium sulphate for fetal neuroprotection reduces the risk of "death or CP" (RR 0.85; 95% CI 0.74 to 0.98; 4 trials, 4446 infants), "death or moderate-severe CP" (RR 0.85; 95% CI 0.73 to 0.99; 3 trials, 4250 infants), "any CP" (RR 0.71; 95% CI 0.55 to 0.91; 4, trials, 4446 infants), "moderate-to-severe CP" (RR 0.60; 95% CI 0.43 to 0.84; 3 trials, 4250 infants), and "substantial gross motor dysfunction" (inability to walk without assistance) (RR 0.60; 95% CI 0.43 to 0.83; 3 trials, 4287 women) at 2 years of age. Results were consistent between trials and across the meta analyses. There is no anticipated significant increase in health care-related costs, because women eligible to receive antenatal MgSO4 will be judged to have imminent preterm birth. VALIDATION: Australian National Clinical Practice Guidelines were published in March 2010 by the Antenatal Magnesium Sulphate for Neuroprotection Guideline Development Panel. Antenatal MgSO4 was recommended for fetal neuroprotection in the same dosage as recommended in these guidelines. However, MgSO4 was recommended only at < 30 weeks' gestation, based on 2 considerations. First, no one gestational age subgroup was considered to show a clear benefit. Second, in the face of uncertainty, the committee felt it was prudent to limit the impact of their clinical practice guidelines on resource allocation. Also in March 2010, the American College of Obstetricians and Gynecologists issued a Committee Opinion on MgSO4 for fetal neuroprotection. It stated that, "the available evidence suggests that magnesium sulphate given before anticipated early preterm birth reduces the risk of cerebral palsy in surviving infants." No official opinion was given on a gestational age cut-off, but it was recommended that physicians develop specific guidelines around the issues of inclusion criteria, dosage, concurrent tocolysis, and monitoring in accordance with one of the larger trials. SPONSORS: Canadian Institutes of Health Research (CIHR). PMID- 21639973 TI - Hypocholesterolemic Effects of N-[(2-Hydroxy-3-N,N-Dimethylhexadecyl Ammonium)Propyl] Chitosan Chloride in High-Fat-Diet-Induced Rats. AB - In this paper, the potential hypocholesterolemic effect of N-[(2-hydroxy-3-N,N dimethylhexadecyl ammonium)propyl] chitosan chloride (N-CQCs) was studied. In vivo hypocholesterolemic tests of N-CQCs were carried out to examine the effect on LDL, HDL, TC, leptin, FFA and SOD. The results showed that LDL, TC, leptin and FFA were decreased after supplementation with N-CQCs in the high-fat diet, preliminarily indicating that N-CQCs have a better hypocholesterolemic effect than chitosan. SOD was significantly elevated by N-CQCs compared with that of HF and HFC groups. N-CQCs increased antioxidant activity of SOD, indicating that N CQCs regulated antioxidant enzyme activities and might reduced lipid peroxidation. PMID- 21639977 TI - Advanced spectral imaging for noninvasive microanalysis of cultural heritage materials: review of application to documents in the U.S. Library of Congress. AB - Hyperspectral imaging was originally developed for remote sensing and astronomical applications, but adaptations of this technology have been of great benefit to the preservation of cultural heritage. Developments in noninvasive analytical techniques have advanced the preservation of cultural heritage materials by enabling the identification and analysis of a range of materials, utilizing their unique spectral response to nondestructively determine chemical composition, and determining states of deterioration and change due to environmental conditions. When used as a tool for noninvasive characterization of cultural heritage, these spectral imaging systems allow the collection of chemical identification information about materials without sampling, which is a critical factor for cultural heritage materials. The United States Library of Congress has been developing the application of hyperspectral imaging to the preservation and analysis of cultural heritage materials as a powerful noncontact technique. It allows noninvasive characterization of materials, by identifying and characterizing colorants, inks, and substrates with narrow-band illumination to protect the object while also monitoring deterioration or changes due to exhibit and other environmental conditions. Contiguous illumination from the ultraviolet, visible, and infrared spectral regions allows the capture of lost, obscured, and deteriorated information. The resulting image cube allows greater capabilities for mapping and coordinating a range of complementary chemical and spectral analyses. The capabilities of this technique are illustrated by a review of results from analysis of the Waldseemuller World Map, the L'Enfant plan for Washington, D.C., and the first draft of the U.S. Declaration of Independence. PMID- 21639978 TI - Multimodal spectral imaging of cells using a transmission diffraction grating on a light microscope. AB - A multimodal methodology for spectral imaging of cells is presented. The spectral imaging setup uses a transmission diffraction grating on a light microscope to concurrently record spectral images of cells and cellular organelles by fluorescence, darkfield, brightfield, and differential interference contrast (DIC) spectral microscopy. Initially, the setup was applied for fluorescence spectral imaging of yeast and mammalian cells labeled with multiple fluorophores. Fluorescence signals originating from fluorescently labeled biomolecules in cells were collected through triple or single filter cubes, separated by the grating, and imaged using a charge-coupled device (CCD) camera. Cellular components such as nuclei, cytoskeleton, and mitochondria were spatially separated by the fluorescence spectra of the fluorophores present in them, providing detailed multi-colored spectral images of cells. Additionally, the grating-based spectral microscope enabled measurement of scattering and absorption spectra of unlabeled cells and stained tissue sections using darkfield and brightfield or DIC spectral microscopy, respectively. The presented spectral imaging methodology provides a readily affordable approach for multimodal spectral characterization of biological cells and other specimens. PMID- 21639979 TI - Time dependence of cellular chemical changes induced in prostate PC-3 cancer cells by two structurally related cardenolides monitored by Fourier transform infrared (FT-IR) spectroscopy. AB - Human PC-3 prostate cancer cells were incubated in the presence of two cardenolides, i.e., ouabain and 19-hydroxy-2''-oxovoruscharin. Their effects were monitored by infrared spectroscopy of the cells after different exposure times to the cardenolides. Analysis of changes in absorbance intensities indicated that, for both compounds, the absorbance at one wavenumber with a minor contribution of a second wavenumber is sufficient to build a linear model accurate enough to assign more than 97% of the spectra to their correct time slot. Student t-tests and twodimensional correlation analysis (2D-COS) indicated that both drugs have very similar effects on PC-3 cells. However, asynchronous 2D maps revealed significant differences and allowed the sequence of the spectral changes to be determined: 1395 -> 1695 cm(-1) for ouabain, and 1400 -> 1655 -> 1100 -> 1250 -> 1020 cm(-1) for 19-hydroxy-2''-oxovoruscharin. 2D correlation map subtraction allowed the identification of very specific differences in the impact of both compounds on PC-3 cells, in particular the ability of 19-hydroxy-2'' oxovoruscharin to affect nucleic acid of PC-3 cells. PMID- 21639980 TI - Comparison between infrared and Raman spectroscopic analysis of maturing rabbit cortical bone. AB - The molecular composition of the organic and inorganic matrices of bone undergoes alterations during maturation. The aim of this study was to compare Fourier transform infrared (FT-IR) and near-infrared (NIR) Raman microspectroscopy techniques for characterization of the composition of growing and developing bone from young to skeletally mature rabbits. Moreover, the specificity and differences of the techniques for determining bone composition were clarified. The humeri of female New Zealand White rabbits, with age range from young to skeletally mature animals (four age groups, n = 7 per group), were studied. Spectral peak areas, intensities, and ratios related to organic and inorganic matrices of bone were analyzed and compared between the age groups and between FT IR and Raman microspectroscopic techniques. Specifically, the degree of mineralization, type-B carbonate substitution, crystallinity of hydroxyapatite (HA), mineral content, and collagen maturity were examined. Significant changes during maturation were observed in various compositional parameters with one or both techniques. Overall, the compositional parameters calculated from the Raman spectra correlated with analogous parameters calculated from the IR spectra. Collagen cross-linking (XLR), as determined through peak fitting and directly from the IR spectra, were highly correlated. The mineral/matrix ratio in the Raman spectra was evaluated with multiple different peaks representing the organic matrix. The results showed high correlation with each other. After comparison with the bone mineral density (BMD) values from micro-computed tomography (micro-CT) imaging measurements and crystal size from XRD measurements, it is suggested that Raman microspectroscopy is more sensitive than FT-IR microspectroscopy for the inorganic matrix of the bone. In the literature, similar spectroscopic parameters obtained with FT-IR and NIR Raman microspectroscopic techniques are often compared. According to the present results, however, caution is required when performing this kind of comparison. PMID- 21639981 TI - Self-aggregation processes of 1,6-diphenyl-1,3,5-hexatriene in water/ethanol mixtures with high water percentages. AB - This work describes the behavior of 1,6-diphenyl-1,3,5-hexatriene (DPH) in ethanol/water mixtures. The dependence of DPH photophysical properties (absorption and fluorescence emission) on the water percentage in ethanol indicates that DPH undergoes self-aggregation processes in solvent conditions above a critical water content. Evidence such as an additional absorption band, Beer's law deviation, kinetic behavior, and other experimental results obtained from temperature variation and surfactant addition demonstrated the presence of several types of DPH aggregates. Resonance light scattering measurements proved that the aggregate grew in water-rich media by a self-catalyzed process. PMID- 21639982 TI - Semi-automated detection of trace explosives in fingerprints on strongly interfering surfaces with Raman chemical imaging. AB - We have previously demonstrated the use of wide-field Raman chemical imaging (RCI) to detect and identify the presence of trace explosives in contaminated fingerprints. In this current work we demonstrate the detection of trace explosives in contaminated fingerprints on strongly Raman scattering surfaces such as plastics and painted metals using an automated background subtraction routine. We demonstrate the use of partial least squares subtraction to minimize the interfering surface spectral signatures, allowing the detection and identification of explosive materials in the corrected Raman images. The resulting analyses are then visually superimposed on the corresponding bright field images to physically locate traces of explosives. Additionally, we attempt to address the question of whether a complete RCI of a fingerprint is required for trace explosive detection or whether a simple non-imaging Raman spectrum is sufficient. This investigation further demonstrates the ability to nondestructively identify explosives on fingerprints present on commonly found surfaces such that the fingerprint remains intact for further biometric analysis. PMID- 21639983 TI - Synthesis and characterization of gold nanoparticles coated with ultrathin and chemically inert dielectric shells for SHINERS applications. AB - We very recently reported a new spectroscopic application for expanding the versatility of surface Raman called "shell-isolated nanoparticle-enhanced Raman spectroscopy" or "SHINERS". The most important and most difficult part of the SHINERS experiment is the effective transfer of the strong electromagnetic field from a gold core through the isolating silica or alumina shell to the probed surface. For this it is essential that the chemically inert dielectric shell be ultrathin (2-5 nm) yet pinhole-free. Herein we describe experimental and theoretical aspects of our SHINERS method in more detail. We provide a protocol for the synthesis and characterization of optimized shell-isolated nanoparticles (SHINs), and we examine the advantages of SHINERS nanoparticles over bare gold nanoparticles. We also present high-quality Raman spectra obtained from gold and platinum single-crystal surfaces in an electrochemical environment by our SHINERS technique. SHINERS is a simple and cost-effective approach that expands the flexibility of surface-enhanced Raman scattering (SERS) for an unprecedented diversity of applications in materials and surface sciences. PMID- 21639984 TI - Design and optimization of the calibration procedure for a miniaturized Fourier transform spectrometer. AB - This paper presents a Fourier transform infrared (FT-IR) spectrometer calibration procedure based on an unusual source made from a spectrally selective surface. An alternative solution to the usual calibrators has been developed to cope with the tight mass budget of an instrument devoted to Mars surface exploration. The designed system has proved effective, in terms of achievable radiometric accuracy, despite the drawbacks due to the significant reflectivity of the sources. The proposed procedure is a standard "two-source" approach in which both cold and hot sources are thermally controlled surfaces, similar to an optical solar reflector, associated to a filament lamp. Such a system allows the required signal to be achieved in the 2-25 l m instrument wavelength range. Source optimization was performed using, as a cost function, the computed radiometric uncertainty, while the required absolute accuracy of the instrument was imposed as the optimization constraint. PMID- 21639985 TI - Back-surface gold mirrors for vibrationally resonant sum-frequency (VR-SFG) spectroscopy using 3-mercaptopropyltrimethoxysilane as an adhesion promoter. AB - Back-surface mirrors are needed as reference materials for vibrationally resonant sum-frequency generation (VR-SFG) probing of liquid-solid interfaces. Conventional noble metal mirrors are not suitable for back-surface applications due to the presence of a metal adhesion layer (chromium or titanium) between the window substrate and the reflective metal surface. Using vapor deposited 3 mercaptopropyltrimethoxysilane (MPTMS) as a bi-functional adhesion promoter, gold mirrors were fabricated on fused silica substrates. These mirrors exhibit excellent gold adhesion as determined by the Scotch((r)) tape test. They also produce minimal spectroscopic interference in the C-H stretching region (2800 3000 cm(-1)), as characterized by VR-SFG. These mirrors are thus robust and can be used as back-surface mirrors for a variety of applications, including reference mirrors for VR-SFG. PMID- 21639986 TI - The detection of palladium particles in proton exchange membrane fuel-cell water by laser-induced breakdown spectroscopy (LIBS). AB - Laser-induced breakdown spectroscopy (LIBS) using conditional data analysis was applied to aqueous suspensions of palladium particles in the reformate water of palladium-based proton exchange membrane fuel cells. A significant amount of palladium was found in the water, indicating degradation of the fuel-cell cathode catalytic layers. The palladium particle-size detection limit was found to be about 400 nm. Calibration procedures to quantify the palladium concentration are discussed. PMID- 21639987 TI - Determination of volume fractions in multicomponent mixtures using ultrasound frequency analysis. AB - Controlling the composition of mixtures is critical for quality control in a wide variety of applications. There is a need for rapid, on-site measurements to optimize processes in real time. Ultrasound easily penetrates opaque samples and containers, yet currently provides minimal chemical information. We have developed a general approach to determine the volume fraction of a liquid in mixtures with multiple components. Ultrasound waves propagating through a medium undergo distortion processes that are characteristic of the chemical bonding composition. The distortion of the waveform can be measured in the ultrasound frequency profile. An ultrasound pulse-through configuration with matching 5 MHz transducers was used to analyze mixtures of water, methanol, and ethanol. Multilinear regression analysis was used to determine the volume fraction of all components in a series of mixtures. Using this technique, volume fractions were determined simultaneously with correlation coefficients (r(2)) greater than 0.98 in two-component mixtures. Determination of volume fractions in three-component mixtures ranging from 65-100% water also showed correlation coefficients of 0.91 for methanol and 0.94 for ethanol. This technique is attractive for process monitoring due to the short measurement time and the simple methodology that excludes sample pretreatment. PMID- 21639988 TI - An advanced digital filter for one-dimensional spectroscopic data: minimizing distortion in band shapes and band intensities. AB - A wide variety of digital filters exist for processing one-dimensional (1D) signals; however, the application of some filters results in pronounced systematic distortions in band shapes and band intensities. In the present contribution, filtering is achieved by optimization in which a general objective function is constructed that possesses a number of desirable qualities, such as (1) smoothness of the resulting spectrum as well as (2) statistical constraints on the residual. Since the residual is explicitly used in the optimization, one can control systematic distortions and therefore avoid over-filtering. In tests using a variety of synthetic as well as real 1D spectroscopic data, the filter adequately preserves both band shapes and band intensities. In addition, the filter appears to accommodate homoscedastic, heteroscedastic, and frequency dependent noise. Examples of its application and usefulness to powder X-ray diffraction (PXRD), Raman, and Fourier transform infrared (FT-IR) emission data are provided. Tests with synthetic data indicate that considerable noise reduction can be achieved in many applications. Finally, an iterative form of the filter is presented. This iterative form further minimizes distortions in band shapes and band intensities when very high levels of denoising are desired. The present filtering approach is an alternative to existing filters, particular when the quality of the residual is important to the user. PMID- 21639989 TI - Automated spectral smoothing with spatially adaptive penalized least squares. AB - A variety of data smoothing techniques exist to address the issue of noise in spectroscopic data. The vast majority, however, require parameter specification by a knowledgeable user, which is typically accomplished by trial and error. In most situations, optimized parameters represent a compromise between noise reduction and signal preservation. In this work, we demonstrate a nonparametric regression approach to spectral smoothing using a spatially adaptive penalized least squares (SAPLS) approach. An iterative optimization procedure is employed that permits gradual flexibility in the smooth fit when statistically significant trends based on multiscale statistics assuming white Gaussian noise are detected. With an estimate of the noise level in the spectrum the procedure is fully automatic with a specified confidence level for the statistics. Potential application to the heteroscedastic noise case is also demonstrated. Performance was assessed in simulations conducted on several synthetic spectra using traditional error measures as well as comparisons of local extrema in the resulting smoothed signals to those in the true spectra. For the simulated spectra, a best case comparison with the Savitzky-Golay smoothing via an exhaustive parameter search was performed while the SAPLS method was assessed for automated application. The application to several dissimilar experimentally obtained Raman spectra is also presented. PMID- 21639990 TI - Detection of copper in water using on-line plasma-excited atomic absorption spectroscopy (AAS). AB - A measurement method and apparatus was developed to measure continuously toxic metal compounds in industrial water samples. The method was demonstrated by using copper as a sample metal. Water was injected into the sample line and subsequently into a nitrogen plasma jet, in which the samples comprising the metal compound dissolved in water were decomposed. The transmitted monochromatic light was detected and the absorbance caused by copper atoms was measured. The absorbance and metal concentration were used to calculate sensitivity and detection limits for the studied metal. The sensitivity, limit of detection, and quantification for copper were 0.45 +/- 0.02, 0.25 +/- 0.01, and 0.85 +/- 0.04 ppm, respectively. PMID- 21639991 TI - Sodium and potassium released from burning particles of brown coal and pine wood in a laminar premixed methane flame using quantitative laser-induced breakdown spectroscopy. AB - A quantitative point measurement of total sodium ([Na](total)) and potassium ([K](total)) in the plume of a burning particle of Australian Loy Yang brown coal (23 +/- 3 mg) and of pine wood pellets (63 +/- 3 mg) was performed using laser induced breakdown spectroscopy (LIBS) in a laminar premixed methane flame at equivalence ratios ( U ) of 1.149 and 1.336. Calibration was performed using atomic sodium or potassium generated by evaporation of droplets of sodium sulfite (Na(2)SO(3)) or potassium sulfate (K(2)SO(4)) solutions seeded into the flame. The calibration compensated for the absorption by atomic alkalis in the seeded flame, which is significant at high concentrations of solution. This allowed quantitative measurements of sodium (Na) and potassium (K) released into the flame during the three phases of combustion, namely devolatilization, char, and ash cooking. The [Na](total) in the plume released from the combustion of pine wood pellets during the devolatilization was found to reach up to 13 ppm. The maximum concentration of total sodium ([Na](max)M(total)) and potassium ([K](max)(total)) released during the char phase of burning coal particles for phi = 1.149 was found to be 9.27 and 5.90 ppm, respectively. The [Na](max)(total) and [K](max)(total) released during the char phase of burning wood particles for phi = 1.149 was found to be 15.1 and 45.3 ppm, respectively. For the case of phi = 1.336, the [Na](max)(total) and [K](max)(total) were found to be 13.9 and 6.67 ppm during the char phase from burning coal particles, respectively, and 21.1 and 39.7 ppm, respectively, from burning wood particles. The concentration of alkali species was higher during the ash phase. The limit of detection (LOD) of sodium and potassium with LIBS in the present arrangement was estimated to be 29 and 72 ppb, respectively. PMID- 21639992 TI - In-situ monitoring of the structure of a goethite-based catalyst during methane oxidation by X-ray absorption near-edge structure (XANES) spectroscopy assisted by chemometric methods. AB - A goethite-based catalyst was evaluated using in-situ X-ray absorption near-edge structure (XANES) spectroscopy during methane oxidation under increasing reaction temperature. Determination of rank by median absolute deviation (DRMAD), evolving factor analysis (EFA), and multivariate curve resolution (MCR) were used to detect the species present in the catalyst during the reaction and determine their concentration profiles and their pure spectra. The reactants and reaction products were monitored on-line by mass spectrometer. The goethite-based catalyst was active for methane oxidation, with CO(2) and H(2)O as the main products. DRMAD and EFA were useful to determine the number of chemical species present in the catalyst structure during reactions. The catalyst presented phase transition during the reaction from goethite to maghemite according to XANES spectra determined by MCR. On the other hand, it was verified that the catalyst presented phase transition from goethite to wustite in the process in the absence of the oxidant (O(2)). PMID- 21639993 TI - Biodegradable nanoparticles for protein delivery: analysis of preparation conditions on particle morphology and protein loading, activity and sustained release properties. AB - PLGA particles have been extensively used as a sustained drug-delivery system, but there are multiple drawbacks when delivering proteins. The focus of this work is to address the most significant disadvantages to the W/O/W double emulsion procedure and demonstrate that simple changes to this procedure can have significant changes to particle size and dispersity and considerable improvements to protein loading, activity and sustained active protein release. A systematic approach was taken to analyze the effects of the following variables: solvent miscibility (dichloromethane (DCM), ethyl acetate, acetone), homogenization speed (10 000-25 000 rpm), PLGA concentration (10-30 mg/ml) and additives in both the organic (sucrose acetate isobutyrate (SAIB)) and aqueous (bovine serum albumin (BSA)) phases. Increasing solvent miscibility decreased particle size, dispersity and protein denaturation, while maintaining adequate protein loading. Increasing solvent miscibility also lowered the impact of homogenization on particle size and dispersity and protein activity. Changes to PLGA concentration demonstrated a minimum impact on particle size and dispersity, but showed an inverse relationship between protein encapsulation efficiency and particle protein weight percent. Most particles tested provided sustained release of active protein over 60 days. Increasing solvent miscibility resulted in increases in the percent of active protein released. When subjected to synthesis conditions with DCM as the solvent, BSA as a stabilizer resulted in the maximum stabilization of protein at a concentration of 100 mg/ml. At this concentration, BSA allowed for increases in the total amount of active protein delivered for all three solvents. The benefit of SAIB was primarily increased protein loading. PMID- 21639994 TI - Combinatorial effects of charge characteristics and hydrophobicity of silk fibroin on the sorption and release of charged dyes. AB - The present study was designed to examine the influence of the charge characteristics of silk fibroin on the sorption and release of charged dyes by varying the pH values of the sorption and release media as well as types of charged dyes. Negatively charged dyes (phenol red and chromotrope 2R) and positively charged dyes (crystal violet and indoine blue) were used as the model compounds. Silk fibroin films were prepared by using a solution casting technique. The prepared films were then treated with an aqueous methanol solution or annealed with water to control their conformation. The sorption behavior of the model compounds made by the methanol-treated and water-annealed silk fibroin films was investigated. Compared to the water- annealed silk fibroin films, a higher hydrophobicity of the methanol-treated silk fibroin films caused a higher sorption of the hydrophobic dyes. The dye molecules had a fairly high affinity to the silk fibroin film, even though the dye and the matrix possessed the same charge. However, in the presence of two charged groups in a single dye molecule, the electrostatic repulsion become more dominant. Stronger interaction was observed when the charges of the film and the dye were opposite. The results of dye sorption and release experiments showed that the degree of synergism or competition between electrostatic and hydrophobic interactions directly depended on the charges and chemical structure of the dye molecules and the environmental pH conditions of the existing silk fibroin film. PMID- 21639995 TI - Fibrous scaffolds made by co-electrospinning soluble eggshell membrane protein with biodegradable synthetic polymers. AB - Soluble eggshell membrane protein (SEP), isolated from natural eggshell membrane, was co-electrospun with biodegradable synthetic polymers poly(propylene carbonate) (PPC) and poly(lactic acid) (PLA) in various proportions from 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP) solutions in order to prepare fibrous scaffolds having simultaneously good mechanical properties and biocompatibility. The fiber morphology was observed by field emission scanning electron microscopy, showing uniform fibers with diameter of 1.2-1.0 and 1.3-0.7 um for PPC/SEP and PLA/SEP blend fibers, respectively. Transmission electron microscopy observation shows that the blend fibers have domain-matrix phase morphology with fiber-like SEP domains in the PPC or PLA matrix, indicating the occurrence of phase separation, although interaction exists between PPC (or PLA) and SEP, as revealed by attenuated total reflectance Fourier transform infrared spectroscopy. The mechanical properties were evaluated by uniaxial tensile tests and showed that both the tensile strength and elongation at break increase with increasing incorporation of PPC (or PLA). The surface composition was investigated by X-ray photoelectron spectroscopy and SEP was found on the fiber surfaces, and as a result the surfaces of the fibrous scaffolds are superhydrophilic. NIH3T3 cell culture tests demonstrate that the PPC/SEP and PLA/SEP blend fibrous scaffolds have a much improved biocompatibility compared to pure PPC or PLA fibrous scaffolds. PMID- 21639996 TI - Outcomes and cost comparison of three therapeutic approaches to allergic rhinitis. AB - BACKGROUND: Allergic rhinitis (AR) is a global health problem because of its increasing impact on economics, society, and the individual's quality of life. This study compares the outcomes and cost of three intranasal therapeutic approaches to the treatment of AR. METHODS: This was a retrospective cohort study using propensity scores to achieve balanced cohorts. The study population included patients >=16 years of age with at least one intranasal prescription claim, without concurrent nasal polyps or sinusitis. Health care use and costs, airway infections, pharmacy costs, and indicators of unsatisfactory treatment (i.e., treatment augmentation or switching) were evaluated in the 1-year follow up period using a claims database. RESULTS: Data from 141,190 patients in intranasal antihistamines (INA) therapy, intranasal steroids (INS) therapy, and intranasal combination therapy (ICT) cohorts were analyzed. The INA cohort showed the lowest rate of change in treatment (switching or augmentation). Switching rates were lowest in the INS therapy cohort, whereas augmentation was lowest in the INA cohort. AR- and asthma-related medication costs were significantly lower in the INA cohort. No differences were observed in airway infections and overall health care costs. Concurrent chronic obstructive pulmonary disorder and asthma were the strongest predictors of health care cost and respiratory infection in the follow-up period. CONCLUSION: A change in treatment was noted in ~1/3 of the entire study population. None of the treatments had a remarkable effect on health care costs or the occurrence of airway infections. The INA treatment cohort had lower AR- and asthma-related medication costs. PMID- 21639997 TI - Human lysozyme has fungicidal activity against nasal fungi. AB - BACKGROUND: The cationic antimicrobial peptide lysozyme is the most prevalent innate immune protein in nasal secretions but there is a paucity of research regarding its role in paranasal sinus disease. Lysozyme is generally regarded as an antibacterial agent; however, some data suggest activity toward yeast. This study was designed to determine if lysozyme displays fungicidal activity toward fungi commonly identified in patients with chronic rhinosinusitis (CRS) or fungal sinusitis. METHODS: Using a colony-forming unit assay the fungicidal activity of lysozyme (0, 0.5, 5, and 50 micromolar; 0- to 7-hour treatment) was tested against strains of Aspergillus fumigatus, the yeast Candida albicans, and other fungi commonly identified in mucin of patients with CRS. Fungi cultured directly from the mucin of two CRS patients were also tested to determine if they were resistant to the fungicidal activity of lysozyme. RESULTS: The fungicidal effect of lysozyme was both concentration and time dependent. After 7-hour treatment lysozyme (5 micromolar) had >80% fungicidal activity against A. fumigatus, Penicillium sp., Acremonium sp., C. albicans, and Candida parapsilosis. The fungicidal activity of lysozyme toward Alternaria alternata could not be determined. Lysozyme was also fungicidal toward the clinical isolates A. fumigatus and Aspergillus terreus cultured from the mucin of CRS patients. CONCLUSION: Lysozyme displays fungicidal activity toward many fungi commonly identified in patients with CRS, as well as clinical fungi isolates cultured from the mucin of CRS patients. Additional studies are required to determine the regulation of lysozyme in CRS. PMID- 21639998 TI - Assessment of palatability of two sublingual diluents in allergic patients: a prospective pilot study. AB - BACKGROUND: Taste of diluents could have an effect on the ultimate patient compliance and acceptability of allergenic extract formulations used in sublingual allergy immunotherapy. This study assessed the palatability and compared the flavor and taste preferences of two sublingual diluents in allergic adult patients. METHODS: This was a prospective, randomized, double-blinded pilot study. Diluents A (50% glycerin and water) and B (0.4% phenol and water) were tested. Three drops were placed in the sublingual area beneath the tongue for 2 minutes before swallowing. A 5-point analog scale was used to allow ranking of the degree of taste acceptance. Patients were then given a 10-minute break where unsalted crackers and bottled water were provided to cleanse the palate between tasting periods. After tasting both samples, patients were asked to answer a final question regarding taste preference. RESULTS: Of the 30 patients, there were 15 men and 15 women with a mean age of 38 years (range, 18 to 61 years). For diluent A, 18 (60.0%) patients graded the diluent as having a "pleasant" taste, 8 (26.7%) patients gave a grade of "very pleasant," and 4 (13.3%) patients said that they were "not sure." For diluent B, 16 (53.3%) patients gave a grade of "unpleasant" and only 6 (20.0%) patients gave a grade of "pleasant." Finally, 28 (93.3%) of the 30 patients chose diluent A over diluent B as the preferred diluent of choice (p < 0.0001). CONCLUSION: Fifty percent glycerin had better taste attributes and phenolated saline induced unpleasant oral sensation on sublingual dosing. PMID- 21639999 TI - The green journal: evolution or revolution? PMID- 21640000 TI - From specialty training to independent practice as a GP. PMID- 21640001 TI - Learning from the expert patient's voice: implications for training. PMID- 21640002 TI - The importance of peer support for new GPs during their transition to independent practice. PMID- 21640003 TI - Primary care for children: back to the future. PMID- 21640004 TI - Reflection on developing an undergraduate course: the value of an action research approach. AB - BACKGROUND: Busy practitioners may not always act on their critical reflection though critical reflection is key to improving practice. This paper presents action research (AR) as a tool that could help to translate reflection into action which can aid improvement of practice. AIM: To critically discuss the development of a medical education course to illustrate the use of AR in enabling reflection on action and improvement to practice in complex educational environments. METHODOLOGY: Developmental evaluation cycles were employed in designing and integrating a cultural diversity course for student doctors. RESULTS: Iterative cycles of evaluation on the cultural diversity course resulted in: (1) increased student centred activities in the course; (2) recognition and attempts to improve student reflective ability around cultural competency issues; (3) opportunity to monitor own teaching practice and; (4) improvement in practice. However, the model described here fell short of a true AR approach because of limited involvement of stakeholders in all stages of the process. CONCLUSION: Future research should investigate AR as a tool for educational change management, postgraduate learning and portfolio documentation. PMID- 21640005 TI - What are the key elements of a primary care teaching practice? AB - This paper explores the perceptions of clinicians, clinical learners and practice staff of key elements of being a teaching practice. Whilst previous studies have explored individual facets of being a teaching practice, this paper offers an in depth insight of how these elements come together to support learning and teaching activity. The single descriptive case study was conducted within a purposefully selected primary care teaching practice in West Yorkshire, UK. Interview, observational and documentary data were collected over a single academic year in 2008/9. Interview data from 33 participants were transcribed and analysed using a modified grounded theory approach. Those interviewed included transient, vocational and embedded medical and nurse learners. The findings are presented within the context of existing literature on teaching practices and practice-based clinical learning. The case study highlighted elements that support and contribute to learning within a practice. These elements include: whole practice support for learning, a skilled and committed clinical and educational workforce, stimulation and support from a range of learners at all levels, and a more indefinable additional element which is best summarised as a passion for education. The findings will be of interest to those involved with the delivery or organisation of clinical learning in primary care. PMID- 21640006 TI - Standardisation of role players for the Clinical Skills Assessment of the MRCGP. AB - This paper describes the processes used to prepare the role players who simulate the 'patients' in the MRCGP Clinical Skills Assessment (CSA) examination, one of the licensing qualifications for GPs completing their training. Because it is such a high-stakes exam, the CSA must be seen to be fair and trustworthy, and such attributes could be compromised by inadequate role player performance. On a CSA examination day, three role players simulate the same 'patient' simultaneously on three different exam floors, each playing the same case up to 26 times on the day, and these 'patients' must present in a consistent and reliable manner to ensure that all candidates face the same test situation. The paper describes the initial selection and training of the actors, how they are prepared on the day of the exam itself, and the quality assurance processes designed to ensure that the exam has excellent role players performing as the simulated 'patients'. PMID- 21640007 TI - Using actors to simulate doctors in the continuing professional development of GP trainers and appraisers. AB - Simulation has radically changed medical training and is set to continue to do so in the future. The branch of simulation that sees actors playing a role in training and assessment is long-established and well-developed. Typically actors have been used to play the role of patient. This innovation in communication skills teaching sees actors playing doctors (specifically GP trainees or GP appraisees), to enhance the continuing professional development of established GP trainers and appraisers. By using actors, scenarios drawing on typical encounters can be played up or down using techniques which are familiar to hi-fidelity electronic simulation. This flexibility enables the 'challenge' of the encounter to be tailored to the needs of the participants in order to maximise their learning. Feedback from participants at the workshops was positive, noting that the approach is both challenging and revealing of their own practice. More research is needed to develop this approach further and explore it's relevance to other areas of GP training and CPD. PMID- 21640008 TI - How to help your international medical graduate trainee pass the CSA. PMID- 21640009 TI - An innovative approach to helping international medical graduates to improve their communication and consultation skills: whose role is it? PMID- 21640010 TI - GP trainer as GP researcher: practicalities of doing a focus group study in primary care. PMID- 21640011 TI - Peer confrontation: a model. PMID- 21640012 TI - Involve patients and carers in training health professionals. PMID- 21640023 TI - AALAS journals reader survey. Survey conducted from 02 to 17 February 2011. Survey results reported to AALAS on 18 February 2011. PMID- 21640024 TI - Hematologic, serologic, and histologic profile of aged Siberian hamsters (Phodopus sungorus). AB - Biologic samples from 18 (12 female, 6 male) Siberian hamsters (Phodopus sungorus) representing an aged colony (17 to 27 mo) were examined. Values for CBC and serum biochemical parameters were determined, and macroscopic and microscopic pathologic evaluations were performed. Blood urea nitrogen levels were significantly higher in male (54.2 +/- 14 mg/dL) compared with female (35.3 +/- 22 mg/dL) hamsters and correlated histologically with a higher incidence of chronic glomerulonephropathy in males (5 of 6 males; 0 of 12 females). All 18 hamsters had histologic evidence of follicular mite infestation. Half (6 of 12) of the female hamsters showed cystic rete ovarii. Other histologic findings included thymic or thyroid branchial cysts (3 of 18), focal enteritis (2 of 18), and single cases of hepatic hemangiosarcoma, renal adenoma, subcutaneous mast cell tumor, cutaneous sebaceous adenoma, cutaneous trichofolliculoma, squamous papilloma of the nonglandular stomach, epididymal cholesteatoma, pyometra, and pituitary craniopharyngeal cyst. This study is the first published report of hematologic and serum chemical values for any population of Siberian hamsters and the first published report showing a potential male predisposition for chronic progressive glomerulonephropathy and a potential female predisposition for cystic rete ovarii. PMID- 21640025 TI - Cardiac tissue Doppler and tissue velocity imaging in anesthetized New Zealand white rabbits. AB - New Zealand white rabbits are commonly used in cardiovascular research. Complete echocardiographic examination of the heart includes the evaluation of tissue Doppler (TDI) parameters, yet normal data are unavailable for rabbits. In addition, tissue velocity imaging (TV) is a potentially useful measure of myocardial function that has not yet been applied to rabbits. Anesthetized New Zealand white rabbits (n = 31) underwent echocardiography to establish the feasibility of performing TDI and TV and establishing corresponding reference values. Standard 2D, M-mode, and Doppler measurements were obtained in all rabbits and showed values comparable to previously published data. Interpretable TDI images were obtained in all 31 rabbits and TV in 24 of 31 rabbits. The values obtained were similar to those seen in healthy cats and are comparable to the values found in adult humans. TDI and TV can easily be added to standard echocardiographic evaluation in rabbits. The values from the current study, obtained in normal rabbits, can be used as reference values to improve characterization of cardiac disease in this species. PMID- 21640026 TI - Spontaneous lesions in aged captive raccoons (Procyon lotor). AB - In nature, free-ranging raccoons typically do not live longer than 2 y; most raccoons in the wild die young due to accidents and diseases. Therefore, few data are available regarding lesions associated with advancing age in raccoons. This communication documents the lesions present in raccoons (7 male; 3 female) that were older than 7 y and had been used as breeders at a commercial facility in central Iowa. The most frequent microscopic lesions in these raccoons included accumulation of iron pigment in livers and spleens (10 of 10 animals evaluated), neuroaxonal degeneration in caudal medulla (10 of 10), vascular mineralization (psammoma body) in choroid plexus (9 of 10), myocardial inclusions (7 of 8), and cystic endometrial hyperplasia (2 of 3). Other conditions were seen with less prevalence. Except for the detection of gastritis with bacteria in the gastric mucosa of 1 raccoon, the presence of inflammatory cells in 3 choroid plexuses, and the presence of Lafora bodies in the brain of 1 animal, all conditions observed had previously been reported in raccoons. Surprisingly, islet-cell amyloidosis, previously observed as common incidental finding in older captive raccoons, was not seen in any of the raccoons we examined. Because free-ranging raccoons are distributed over wide geographic areas, their local environment may have considerable influence on the range of spontaneous lesions that would occur in raccoons obtained from a specific location. Therefore, the lesions found in these raccoons from central Iowa may differ from those of other raccoon populations. PMID- 21640027 TI - Eliminating animal facility light-at-night contamination and its effect on circadian regulation of rodent physiology, tumor growth, and metabolism: a challenge in the relocation of a cancer research laboratory. AB - Appropriate laboratory animal facility lighting and lighting protocols are essential for maintaining the health and wellbeing of laboratory animals and ensuring the credible outcome of scientific investigations. Our recent experience in relocating to a new laboratory facility illustrates the importance of these considerations. Previous studies in our laboratory demonstrated that animal room contamination with light-at-night (LAN) of as little as 0.2 lx at rodent eye level during an otherwise normal dark-phase disrupted host circadian rhythms and stimulated the metabolism and proliferation of human cancer xenografts in rats. Here we examined how simple improvements in facility design at our new location completely eliminated dark-phase LAN contamination and restored normal circadian rhythms in nontumor-bearing rats and normal tumor metabolism and growth in host rats bearing tissue-isolated MCF7(SR(-)) human breast tumor xenografts or 7288CTC rodent hepatomas. Reducing LAN contamination in the animal quarters from 24.5 +/- 2.5 lx to nondetectable levels (complete darkness) restored normal circadian regulation of rodent arterial blood melatonin, glucose, total fatty and linoleic acid concentrations, tumor uptake of O(2), glucose, total fatty acid and CO(2) production and tumor levels of cAMP, triglycerides, free fatty acids, phospholipids, and cholesterol esters, as well as extracellular-signal-regulated kinase, mitogen-activated protein kinase, serine-threonine protein kinase, glycogen synthase kinase 3beta, gamma-histone 2AX, and proliferating cell nuclear antigen. PMID- 21640028 TI - Effect of sampling strategy on the detection of fur mites within a naturally infested colony of mice (Mus musculus). AB - Fur mites are one of the most common ectoparasites of laboratory mice and traditionally are diagnosed through surveillance of individual colony animals. Although multiple diagnostic modalities exist, few recommendations suggest optimal testing methods, target colony populations, or sampling sites. We compared the fur pluck and sticky paper techniques for the diagnosis of Myocoptes musculinus in naturally infested immunocompetent mice and evaluated the effect of mouse age and sampling site on the efficacy of fur plucks. We found that the sticky paper technique was more likely to detect fur mites than were fur plucks. Housing mice individually increased the incidence of false-negative fur pluck tests, whereas sensitivity was equivalent for preweanling and adult mice. The ventral abdomen was the most likely single sampling location to detect evidence of any stage of Myocoptes musculinus, but fur mite eggs were overrepresented on the neck. We found that the surface temperature of the murine neck surface was warmer than was the rump and therefore may represent a unique microenvironment for fur mite egg development. Given our findings, we recommend that group-housed adult or preweanling mice should be selected for Myocoptes musculinus evaluation and that the ventral abdomen should be sampled. When possible, the postmortem sticky paper technique should be used rather than the antemortem fur pluck method. PMID- 21640029 TI - Effect of a short-term fast on ketamine-xylazine anesthesia in rats. AB - Although ketamine-xylazine (KX) anesthesia is commonly used in rats, it is often reported to have an inconsistent anesthetic effect, with a prolonged induction time, an inadequate anesthetic plane, or a very short sleep time. Blood flow to the liver is known to shift after a meal in rats, perhaps explaining anesthetic variability among rats with variable prandial status. The current study tested the hypothesis that a short period of fasting (3 h) prior to induction with intraperitoneal KX anesthesia would provide a shorter time to recumbency, a longer total sleep time, and a more consistent loss of toe pinch response than would fed rats. Two groups of male Sprague-Dawley rats were used in blinded, crossover experiments. KX anesthesia was administered at 2 different doses (50 mg/kg-5 mg/kg and 70 mg/kg-7 mg/kg) after ad libitum feeding or a 3-h fast. There were no significant differences between groups in induction time, total sleep time, or loss of toe pinch response. We conclude that fasting rats for 3 h prior to KX intraperitoneal anesthesia does not affect induction time, total sleep time, loss of toe pinch response or reduce KX anesthetic variability in male Sprague-Dawley rats. PMID- 21640031 TI - Analgesic effects of meloxicam, morphine sulfate, flunixin meglumine, and xylazine hydrochloride in African-clawed frogs (Xenopus laevis). AB - We evaluated analgesic use and analgesiometry in aquatic African-clawed frogs (Xenopus laevis). We used the acetic acid test (AAT) to assess the analgesic potential of systemic xylazine hydrochloride, meloxicam, flunixin meglumine, and morphine sulfate after injection into the dorsal lymph sac. Flunixin meglumine provided better analgesia than did the other drugs, most evident at 5 and 9 h after administration. Because the AAT was associated with the development of dermal lesions, we discontinued use of this assay and chose the Hargreaves test as an alternative method of measuring nociception in Xenopus. This assay is commonly performed in rodents, but its efficacy in an aquatic species such as Xenopus was unknown prior to this study. We found that the Hargreaves test was an effective measure of nociception in Xenopus, and we used it to evaluate the effectiveness of the nonopiod agents xylazine hydrochloride, meloxicam, and flunixin meglumine both in the absence of surgery and after surgical oocyte harvest. Similar to findings from the AAT, flunixin meglumine provided better analgesia in the Hargreaves test than did the other agents when analyzed in the absence of surgical intervention. Results were equivocal after oocyte harvest. Although surgical oocyte harvest is a common procedure in Xenopus, and currently there are no published recommendations for analgesia after this invasive surgery. Future studies are needed to clarify the efficacy of nonsteroidal antiinflammatory drugs for that purpose. PMID- 21640030 TI - High doses of ketamine-xylazine anesthesia reduce cardiac ischemia-reperfusion injury in guinea pigs. AB - Choosing an appropriate anesthetic protocol that will have minimal effect on experimental design can be difficult. Guinea pigs have highly variable responses to a variety of injectable anesthetics, including ketamine-xylazine (KX). Because of this variability, supplemental doses often are required to obtain an adequate plane of anesthesia. Our group studies the isolated guinea pig heart, and we must anesthetize guinea pigs prior to harvesting this organ. In this study, we sought to determine whether a higher dose of KX protected isolated guinea pig hearts against myocardial ischemia-reperfusion injury. Male Hartley guinea pigs (Crl:HA; 275 to 300 g; n = 14) were anesthetized with either of 2 doses of KX (K: 85 mg/kg, X: 15 mg/kg; or K: 200 mg/kg, X: 60 mg/kg). After thoracotomy, hearts underwent 20 min of ischemia followed by 2 h of reperfusion. The high dose of KX significantly reduced myocardial infarct size as compared with the low dose (36% +/- 3% and 51% +/- 6%, respectively). Furthermore, the high dose of KX improved hemodynamic function over that associated with the low dose as measured by increases in both left ventricular developed pressure (49 +/- 4 and 30 +/- 8 mm Hg, respectively) and maximal rate of left ventricular relaxation (-876 +/- 70 and -576 +/- 120 mm Hg/s, respectively). However, the high dose of KX did not alter the maximal rate of left ventricular contraction or coronary flow. These results suggest that supplementation of KX to ensure an adequate anesthetic plane may introduce unwanted variability in ischemia-reperfusion studies. PMID- 21640032 TI - Intestinal resection and anastomosis in neonatal gnotobiotic piglets. AB - We describe a surgical method for ileal resection and anastomosis in newborn germfree piglets that was undertaken to establish a model that can be used for immunologic research and other applications. A preliminary experiment indicated that neonatal piglets with resection of approximately 60 cm of their ileum (removal of approximately 90% of the continuous ileal Peyer patches; group A) and those in which the ileum was transected (group B) could be maintained germfree for 35 d, colonized with defined gut flora, and maintained in a clean room until 70 d of age. In the final study, 12 piglets (4 each for groups A and B and 4 untreated controls), were monitored for postoperative feeding behavior, malaise, evidence for contamination with pathogenic bacteria, and weight gain. All surgical animals were free from incidental contamination from pathogens and environmental organisms with atypical colony types for 35 d. Two piglets in group B died postoperatively (1 during the preliminary experiment and 1 during the final study). Control (group C) piglets gained significantly more weight than did those in group A. These studies demonstrated that surgical resection of the ileal Peyer patches under germfree conditions can be accomplished successfully without compromising piglet health or introducing pathogens and with only a modest reduction in weight gain. PMID- 21640033 TI - Performance and longevity of a novel intraosseous device in a goat (Capra hircus) model. AB - We performed 2 studies to assess the function and longevity of a novel intraosseous catheter device. For study 1, 9 goats were assigned to 3 groups (intraosseous catheter in the proximal humerus, intraosseous catheter in the proximal tibia, or standard jugular catheter). Devices in the tibia remained in place for less time than did those in the humerus, and no goats exhibited radiographic evidence of resulting damage or structural change in surrounding bone. Positive bacterial cultures were found in all 9 goats at various time points. In study 2, 18 goats were assigned to 2 groups (intraosseous catheter in the wing of the ilium or proximal humerus). Samples for serial aerobic and anaerobic blood cultures and CBC were collected while devices remained in use. Clinical monitoring and removal criteria were identical those for study 1. Catheters in the ilium remained in place for less than 24 h on average, and those in the humerus remained in place for an average of 2.5 d. Several goats with proximal humeral catheters demonstrated moderate lameness after removal, and radiographic evidence of periosteal bone growth was noted in another goat. Bloodwork indicated mild elevations of WBC counts from baseline in some cases. Bacterial growth was found in samples from 4 of 18 goats at various time points. Our study indicated that intraosseous catheters may remain safely in place for more than 24 h, but animals should be monitored closely for negative side effects for several days after removal. PMID- 21640034 TI - Effects of aging and blood contamination on the urinary protein-creatinine ratio in captive chimpanzees (Pan troglodytes). AB - The initial goal of this study was to evaluate proteinuria by using the protein to creatinine (UPC) ratio of urine obtained by cystocentesis of healthy adult captive chimpanzees. Urine samples were collected by using ultrasound-guided cystocentesis from 125 (80 male, 45 female) captive chimpanzees. All samples were collected over a 17-mo time period (August 2008 to January 2010) during the animal's annual physical examination. Samples were assayed at a veterinary diagnostic laboratory. Results indicated that both age and blood contamination affect the UPC ratio and therefore alter the diagnostic utility of the UPC ratio in chimpanzees. In addition, this research establishes reference ranges by age for the UPC ratio in healthy adult chimpanzees. Chimps younger than the median age of 24.6 y have a median UPC ratio of 0.098 (range, 0 to 1.76), whereas older animals have a median UPC of 0.288 (range, 0 to 2.44). Our results likely will enable veterinarians working with chimpanzees to better evaluate their renal function. PMID- 21640035 TI - Strategies to prevent, treat, and provoke Corynebacterium-associated hyperkeratosis in athymic nude mice. AB - Athymic nude mice infected with Corynebacterium bovis typically exhibit transient hyperkeratotic dermatitis. Our vivarium experienced an increased incidence of disease characterized by persistent skin lesions and increased mortality, leading to this study. For detection of infection, skin and buccal swab methods showed comparable sensitivities in nude mice. Various prevention, treatment, and eradication strategies were evaluated through clinical assessment, microbiology, and histopathology. In experimentally naive athymic nude mice, a 2-wk course of prophylactic amoxicillin-containing diet (1200 ppm amoxicillin; effective dose, 200 mg/kg) was ineffective at preventing infection or disease. There was also no significant difference in disease duration or severity in athymic nude mice that received amoxicillin diet or penicillin-streptomycin topical spray (penicillin, 2500 U/mL; streptomycin, 2500 MUg/mL). Prolonged treatment with 4 or 8 wk of amoxicillin diet cleared only a small number of athymic nude mice that had subclinical C. bovis infections. Antibiotic sensitivity of C. bovis isolates demonstrated a small colony isolate with less susceptibility to all antibiotics compared with a large colony isolate. Resistance did not appear to develop after prolonged treatment with amoxicillin. Provocation testing by administration of cyclophosphamide (50 mg/kg i.p. every 48 to 72 h for 90 d) to subclinically infected athymic nude mice resulted in prolonged clinical disease that waxed and waned without progression to severe disease. Our findings suggest that antibiotic prophylaxis and treatment of clinical disease in experimentally naive mice is unrewarding, eradication of bacterial infection is difficult, and severe disease associated with C. bovis is likely multifactorial. PMID- 21640036 TI - Pharmacokinetics of cefovecin in cynomolgus macaques (Macaca fascicularis), olive baboons (Papio anubis), and rhesus macaques (Macaca mulatta). AB - Cefovecin sodium is a long-acting, third-generation, cephalosporin antibiotic approved for the treatment of skin infections in dogs and cats. The pharmacokinetic properties of cefovecin were evaluated in cynomolgus macaques (Macaca fascicularis), olive baboons (Papio anubis), and rhesus macaques (Macaca mulatta) by using a single-dose (8 mg/kg SC) dosing regimen. Plasma cefovecin concentrations were determined by using ultra-performance liquid chromatography with tandem mass spectrometry, and a noncompartmental model was used to determine pharmacokinetic parameters. The half-life of cefovecin was 4.95 +/- 1.47 h in cynomolgus macaques, 9.17 +/- 1.84 h in olive baboons, and 8.40 +/- 2.53 h in rhesus macaques. These values are considerably lower than the half-lives previously published for dogs (133 h) and cats (166 h). The extended half-life of cefovecin in dogs and cats is speculated to be due to active reabsorption of drug in the kidney tubules because plasma clearance is well below the normal glomerular filtration rate. In nonhuman primates, renal clearance rates approximated plasma clearance rates, suggesting that active renal reabsorption of cefovecin does not occur in these species. The pharmacokinetic properties of cefovecin in nonhuman primates are vastly different from the pharmacokinetic properties in dogs and cats, precluding its use as a long-acting antibiotic in nonhuman primates. This study highlights the importance of performing pharmacokinetic studies prior to extralabel drug usage. PMID- 21640037 TI - Hyperhidrosis in naive purpose-bred beagle dogs (Canis familiaris). AB - This case study details the unusual clinical findings in a unique paw-pad disorder that recently emerged among 2 male and 1 female naive purpose-bred beagle dogs (Canis familiaris) newly received into our facility. During acclimation period physical examinations, the affected dogs demonstrated constantly moist, soft paw pads on all 4 feet. No information was available regarding the epidemiology and pathogenesis of this pad condition in beagle dogs. Here, we report the results of physical examination, clinical chemistry analysis, hematology, histopathology, detailed observations, and novel testing techniques performed during the acclimation period. Histopathology of several sections of affected footpads was compared with that of an age-matched dog with clinically normal paw pads. We describe the morphologic features of a distinctive cutaneous canine footpad condition and discuss the possible differential diagnoses. The histologic and clinical features were most consistent with those of hyperhidrosis; to our knowledge, this report is the first description of hyperhidrosis as a distinct condition in purpose-bred beagle dogs. PMID- 21640038 TI - Characterization of spontaneous subclavian steal phenomenon in a female rhesus macaque (Macaca mulatta). AB - In subclavian steal phenomenon (SSP), the subclavian artery develops a stenoocclusive disease proximal to the origin of the vertebral artery, leading to pronounced hemodynamic changes such as arterial flow reversal. Although SSP is a common echographic finding in humans, the phenomenon occurs only rarely in animals; consequently its physiologic features have not been reported previously. Here we describe the clinical and morphologic features of a spontaneous left SSP that was an incidental finding in an 18-y-old female rhesus macaque (Macaca mulatta). Our findings were documented through high-quality imaging studies obtained by using a computerized 3D tomography apparatus and clinical assessment of systolic and diastolic blood pressures. PMID- 21640039 TI - Spontaneous primary squamous cell carcinoma of the lung in a rhesus macaque (Macaca mulatta). AB - A 3-y-old male rhesus macaque (Macaca mulatta) was noticed to be lethargic in the compound. Physical exam revealed cyanotic mucous membranes, dyspnea, bilateral harsh lung sounds, wheezing on expiration, and a firm mass possibly associated with the liver. Radiographs revealed bilateral soft tissue opacities in the thorax. Due to poor prognosis, the rhesus was euthanized, and a necropsy was performed. Both right and left lung lobes were consolidated and had multifocal white-tan masses. On cut section, the masses were firm, had areas of necrosis, hemorrhage, and often contained a tenacious exudate. Masses were identified in the liver and both kidneys. Given the morphologic features of the neoplasm, a diagnosis of squamous cell carcinoma was made. Immunohistochemistry staining for thyroid transcription factor, a nuclear transcription factor normally found in lung, thyroid, and tumors arising from either of those tissues, confirmed that the masses originated from the lung. Malignant primary lung tumors are divided into 8 main histologic subtypes: squamous cell carcinoma, small-cell carcinoma, large-cell carcinoma, adenocarcinoma, adenosquamous carcinoma, sarcomatoid carcinoma, carcinoid tumor, and salivary gland tumors. Clinical signs associated with lung tumors include, but are not limited to, dyspnea, coughing, hemoptysis, lethargy, anorexia, and weight loss. Although squamous cell carcinoma will be low on the differential list for these clinical signs, we encourage clinicians and researchers to not rule it out solely based on incidence and age of the animal. PMID- 21640040 TI - Burden of depressive symptoms in South African public healthcare patients with established rheumatoid arthritis: a case-control study. AB - OBJECTIVES: The burden of depressive symptoms and how demographic and disease characteristics relate to depressive symptoms in patients with rheumatoid arthritis (RA) that belong to developing populations, are currently unknown and were therefore assessed in a case-control study in public healthcare patients in South Africa, a lower-middle income country. Public healthcare attendance is a surrogate of belonging to the developing population in South Africa. METHODS: Demographic and RA features were recorded in 441 public and 202 private healthcare patients. The outcome characteristic was the Arthritis Impact Measure Scales (AIMS) depression score. Relationships of patient characteristics and public healthcare attendance with depressive symptoms were determined in multivariable regression models. RESULTS: The mean +/- SD AIMS depression score was 3.6+/-2.1 and 2.3+/-1.7 in public and private healthcare patients, respectively (p<0.0001 before and after adjustment for covariates). Physical disability was associated with depressive symptoms in both healthcare sectors. Other characteristics that were related to depressive symptoms comprised younger age, male sex and pain in public healthcare patients and fatigue and non-use of disease modifying agents in private healthcare patients. In all patients, public healthcare attendance (standardised beta [95% CI]=0.22 [0.12, 0.32], p<0.0001) and physical disability (standardised beta [95% CI]=0.25 [0.16, 0.34], p<0.0001) were most strongly associated with depressive symptoms. CONCLUSIONS: The burden of depressive symptoms is markedly enhanced in our developing population with RA, independent of age, sex, ethnic origin and disease characteristics. In this setting, the role of social factors should be assessed and, despite restricted resources, depressive symptoms should be routinely addressed. PMID- 21640041 TI - The psychological defensive profile of primary Sjogren's syndrome patients and its relationship to health-related quality of life. AB - OBJECTIVES: We aimed to assess the defensive profile of primary Sjogren's syndrome (SS) patients and to investigate the independent associations of psychological distress and personality variables with health-related quality of life (HRQOL). METHODS: In 40 primary SS patients we assessed psychological distress (SCL-90-R), ego defense mechanisms (Defense Style Questionnaire), hostility features (HDHQ) and HRQOL (WHOQOL-BREF). Fifty-six patients with Systemic Lupus Erythematosous (SLE) and 80 healthy participants matched for age and sex served as controls. RESULTS: Primary SS patients presented higher rates of general psychological distress compared to SLE and healthy participants. Symptoms of somatisation were more prominent in SS than SLE or healthy controls. SS patients presented less use of humour defense and more help-rejecting complains and delusional guilt hostility, compared to controls. Primary SS patients' HRQOL was more impaired than healthy participants and comparable to SLE. Psychological distress was a constant independent correlate of SS patients' HRQOL, while less use of humour (p<0.001) and higher rates of delusional guilt (p=0.032) were also significantly associated with Physical HRQOL independently of psychological distress; more use of schizoid fantasy was also independently associated with impaired Environment HRQOL (p=0.005). CONCLUSIONS: Primary SS patients exhibit several specific psychological difficulties in adaptation to life stressors, and clinicians and consultation-liaison psychiatrists, apart from the early assessment and treatment of psychological distress and somatisation symptoms, should consider the patients' underlying defensive profile and coping capacities, since such personality traits, although usually underestimated, are also independently associated with the disease outcome. PMID- 21640042 TI - A double-blind randomized controlled trial appraising the symptom-modifying effects of colchicine on osteoarthritis of the knee. AB - OBJECTIVES: Osteoarthritis (OA) is the most common articular disease. Common OA treatments are either not effective or associated with side effects. Calcium containing crystals are quite common in primary OA and they worsen or may cause OA through induction of inflammation by neutrophils. Colchicine inhibits urate crystal and calcium-pyrophosphate (CPP) crystal induced inflammation and elastase, a matrix-metalloproteinase (MMP) that play a pivotal role in degenerative joint processes. Hence, it was hypothesized that it may have symptom modifying effects on OA. METHODS: Sixty-one postmenopausal patients with primary knee OA were enrolled. None of them had joint involvement atypical for OA or evidences of chondrocalcinosis in radiographic studies suggesting the presence of calcium-pyrophosphatedeposition-disease (CPPD). Participants were allocated to two groups receiving 0.5mg colchicines BID or placebo. Both groups received common OA treatments. Acetaminophen less than 2gr/day was used as rescue analgesic. The efficacy end points were: patients' global assessment and physician's global assessment, recorded on a VAS (visual analogue scale). Statistical analysis was performed 3 months later. RESULTS: Thirty-one patients were assigned to the colchicine group. Fifty-eight patients were present for the last survey. Only 1 patient in colchicine group encountered adverse effect of colchicine without significant difference between the two groups. Acetaminophen consumption was significantly less in the colchicine (879.3+/-369.7) compared to placebo group (1620.7+/-393.1, p=0.000). Improvement rate at the end of 3 months was significantly higher in the colchicine group for both patients' global assessment and physician's global assessment measures compared to placebo group, (11.14+/-4.06 vs. 3.14+/-2.18, p=0.000) and (9.83+/-3.799 vs. 3.72+/-3.35, p=0.000), respectively. CONCLUSIONS: The efficacy and safety of colchicine for pain reduction in OA was affirmed by our double-blind randomised controlled trial. PMID- 21640043 TI - Baseline characteristics of the population enrolled in the Italian Observational Study on Severe Osteoporosis (ISSO). AB - OBJECTIVES: Baseline characteristics of the population enrolled in the ISSO study, designed to evaluate the incidence of vertebral and non-vertebral fractures in Italian patients with severe osteoporosis treated according to clinical practice over 24 months observation. METHODS: Prospective observational study in 783 post-menopausal women and men entering 18-month treatment with teriparatide in a community setting at 57 centres in Italy. Characterisation included demographics, fracture risk factors, bone mineral density, fracture status, Health-Related Quality of Life (HRQoL) measured by the European Quality of Life Questionnaire, EQ-5D, and back pain assessed by VAS. RESULTS: Most patients were elderly women (90.5%), mean age+/-SD was 72.9+/-8.8 years. Nearly all (91.3%) had experienced >= 1 vertebral fracture (mean+/-SD, 3.6+/-2.2 per patient), 37.5% had >= 1 non-vertebral fracture (mean+/-SD, 1.4+/-0.7 per patient). Nearly all patients were suffering from back pain (94.9%), which had significantly restricted their daily activities (51.7%) and had likely or very likely been caused by vertebral fractures (29.2% and 55.8%, respectively). Mean EuroQoL EQ-5D index value was 0.58+/-0.25 and VAS score 49.2+/-23.6. Non vertebral fractures, back pain and multiple vertebral fractures were associated with lower HRQoL (EuroQoL-5D Index both p<0.001, EQ-5D VAS score p=0.025 and p<0.016, respectively). Many patients were physically inactive (81.1%). One third (34.7%) of population had co-morbidities and 60.5% were on chronic concomitant treatments. Few subjects reported a maternal history of osteoporosis (15.5%), regular consumption of alcohol (13.3%) or were current smokers (11.5%). Nearly two-thirds (71.5%) had already been treated for osteoporosis, mainly with bisphosphonates. Calcium and vitamin D supplements were taken by 13% and 15.5% of the total population, respectively. CONCLUSIONS: At enrollment, the population of ISSO study mostly consisted in aging women, who had osteoporosis with high fracture risk, poor HRQoL and suffered from significant back pain. Most of them had already been treated by bisphosphonates but without calcium and vitamin D supplements. Back pain, as well as non-vertebral and multiple vertebral fractures, were associated with lower HRQoL. PMID- 21640044 TI - Anti-citrullinated peptide antibodies and the progression of radiographic joint erosions in patients with early rheumatoid arthritis treated with FIN-RACo combination and single disease-modifying antirheumatic drug strategies. AB - OBJECTIVES: To evaluate the impact of antibodies to cyclic citrullinated peptide (ACPAs) on radiographic progression in patients with early rheumatoid arthritis (RA) initially treated either with a combination of 3 disease-modifying antirheumatic drugs (DMARDs) or with a single DMARD. METHODS: This study included 129 patients with early active RA initially randomised to treatment either with a combination of methotrexate, sulfasalazine, hydroxychloroquine, and prednisolone (FIN-RACo) (n=69) or with a single DMARD (initially sulfalasalazine) with or without prednisolone (SINGLE) (n=60). After 2 years, the use of DMARDs and prednisolone became unrestricted. Radiographic progression in hands and feet was assessed at baseline and at 1, 2, 3, 4 and 5 years. ACPAs at baseline were determined with enzyme immunoassay. RESULTS: ACPAs were positive in 92 (71%) patients. ACPA-positive vs. negative patients were more frequently rheumatoid factor (RF) positive (83% vs. 22%, p<0.001) and had an erosive disease (54% vs. 22%, p<0.001) at baseline. The presence of ACPA was associated with radiographic progression in FIN-RACo group even when the impact of RF was controlled; the radiographic progression was remarkably slower in ACPA-negative than in ACPA positive cases (RF adjusted change over time between groups p=0.034). In the SINGLE group, the radiographic changes progressed parallel in ACPA-negative and positive patients. CONCLUSIONS: Most ACPA-positive RA patients have joint erosions already at diagnosis. ACPA positivity in early RA was related to radiographic progression even in patients treated initially with the FIN-RACo regimen. The initial FIN-RACo therapy seems to slow down the progression of joint damage in ACPA-negative patients. PMID- 21640045 TI - Association of interleukin-2, interleukin-4 and transforming growth factor-beta gene polymorphisms with Behcet's disease. AB - Behcet's disease (BD) is a chronic immune-mediated disease, characterised by oral and genital lesions and ocular inflammation. As cytokines seem to have important roles in the pathogenesis of BD and production of cytokines could be affected by genetic polymorphisms, this study was performed to investigate gene polymorphisms of a number of cytokines in the patients with BD in comparison with control subjects. One hundred and fifty patients with BD were enrolled in this study. Interleukin (IL)-2 (-330, +166), IL-4 (-1098, -590, -33), IL-10 (-1082, -819, 592), IL-12 (-1188), IFN-gamma (5644), transforming growth factor (TGF)-beta (codon 10, 25), and IL-4RA (+1902) typing were performed by polymerase chain reaction with sequence-specific primers. In the patients with BD, there were significantly increased frequency of IL-2 (-330) GG genotype (p<0.001), IL-4 ( 33) CC genotype (p<0.001), and TGF-beta (codon 10) CC genotype (p=0.004). Meanwhile a significant decrease in the frequency of IL-4 (-33) TC genotype (p<0.001) was detected in the patient group in comparison with normal controls. The genotype CC of TGF-beta at codon 10 was also significantly overrepresented in the patient group (p=0.004). Haplotype frequencies of IL-4 (-1098, -590, -33) showed that the frequency of TTC haplotype was significantly increased in the patients (p<0.001), whereas TTT haplotype was significantly decreased in this group of patients (p<0.001). There was not any significant difference in allele and genotype frequencies of IL-10, IL-12, IFN-gamma, and IL-4RA between patient and control groups. Cytokine single nucleotide polymorphisms could play a role in the pathophysiology of BD. The results of this study could suggest a tendency towards higher production of IL-2 and lower production of IL-4 in the patients with BD. PMID- 21640046 TI - Is headache in Behcet's disease related to silent neurologic involvement? AB - OBJECTIVES: Headache is an interesting issue in Behcet's disease (BD). This study aimed to investigate if headache or a special type of headache was correlated with silent neurologic involvement in BD patients without any neurologic sign. METHODS: The study was performed on 120 BD patients (30 without headache, 30 with non-structural headache of BD and 30 with migraine headache, 30 with tension type headache) and 30 healthy control subjects. Some neurophysiologic tests of brain stem; temporalis muscle exteroceptive suppression periods (ESP) and brain stem auditory evoked potentials (BAEP) were performed in the patients, when they were not in an attack period of the disease, and control subjects to investigate the presence of silent neurologic involvement and the relation between headache and silent neurologic involvement. RESULTS: Some electrophysiological abnormalities, as right BAEP 1-5 interpeak latency prolongation (p=0.01) and left ESP2 duration shortening (p<0.005), were seen in BD patients compared to healthy control subjects. Furthermore, the patients with non-structural headache of BD were found to have shorter ESP1 and 2 durations (p<0.001) and longer ESP1 latencies (p<0.05), with respect to the other patient subgroups with different types of headache and healthy control group, showing brain stem pathology. Additionally, they had longer right BAEP 3-5 interpeak latency as compared to the patient subgroup without headache (p=0.001). CONCLUSIONS: There is a silent neurologic involvement in BD and this involvement may be in relation with a particular type of vascular headache, named as non-structural headache of BD. So, in clinical evaluation of BD patients, this type of headache may be considered as a warning message for neurological involvement. PMID- 21640047 TI - Body composition phenotypes in systemic lupus erythematosus and rheumatoid arthritis: a comparative study of Caucasian female patients. AB - OBJECTIVES: The amount and distribution of fat and lean mass have important implications for health and systemic inflammation may represent a risk for altered body composition. The aim of this study was to analyse whether changes in body composition are similarly associated with systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA), two inflammatory conditions of different pathogenesis. METHODS: Body mass index (BMI), waist circumference, fat mass (FM) and fat-free mass (FFM) were measured in 92 women with SLE, 89 with RA and 107 controls. Results were compared among the 3 groups and correlations of FM percentage were explored within SLE and RA. RESULTS: Abnormal body composition was more frequent in women with SLE and RA than in non-inflammatory controls, despite having a similar BMI. RA diagnosis was significantly associated with overfat (OR=2.782, 95%CI 1.470-5.264; p=0.002) and central obesity (OR=2.998, 95%CI 1.016-8.841; p=0.04), while sarcopenia was more common among SLE (OR=3.003; 95%CI 1.178-7.676; p=0.01). Sarcopenic obesity, i.e. the coexistence of overfat with sarcopenia, was present in 6.5% of SLE and 5.6% of RA women, but no controls. Independent correlations of FM percentage in women with SLE included smoking, disease activity and CRP. In RA, education, disease activity and cumulative corticosteroid dose were identified as independent predictors of FM percentage. CONCLUSIONS: Women with SLE or RA diagnosis are more likely to have abnormal body composition phenotype, with some differences existing between these two conditions. Changes in body composition are partly explained by the inflammatory burden of disease and its treatment. PMID- 21640048 TI - Adjusted prophylactic doses of nadroparin plus low dose aspirin therapy in obstetric antiphospholipid syndrome. A prospective cohort management study. AB - OBJECTIVES: Current guidelines for the treatment of patients with obstetric antiphospholipid syndrome (APS) recommend low dose aspirin (LDA) and prophylactic doses of low molecular weight heparin (LMWH). Most clinicians use a fixed dosage of LMWH in pregnant APS women despite the fact that there are no clinical trials establishing that fixed doses are more efficacious than adjusted ones in preventing pregnancy complications. The efficacy and safety of adjusted single daily doses of LMWH (nadroparin) combined with LDA have thus been evaluated in 33 consecutive pregnancies in women with diagnosed obstetric APS. METHODS: LMWH doses were augmented as the pregnancies progressed and maternal/foetal weight increased. 70-80-90 U/Kg doses ranging between 3800 and 6650 U were administered daily during the first, second and third trimesters, respectively. LDA (100 mg/day) was also prescribed. RESULTS: Pregnancy outcome was successful in 97% of the patients studied, who delivered, between the 29th and 41st weeks of gestation (mean 37.4 +/-2.1 SD), 32 infants with a mean birth weight of 3084 g +/- 514 SD. One woman (3%) experienced a spontaneous abortion at the 8th week of gestation. CONCLUSIONS: The high live birth rate, the satisfactory mean gestational age and weight at birth and the absence of major pregnancy/neonatal-associated complications indicate that adjusted, once daily doses of LMWH together with LDA could be an efficacious treatment option for pregnant APS patients with no history of thrombosis. PMID- 21640049 TI - Synovial colony-stimulating factor-1 mRNA expression in diffuse pigmented villonodular synovitis. AB - OBJECTIVES: To delineate the molecular mechanisms underlying the process of the diffuse-type giant cell tumours, also called pigmented villonodular synovitis, a rare, aggressive condition of the synovium, the knee synovial tissue expression of colony-stimulating factor-1 gene, as detected by real-time polymerase chain reaction, was compared between patients affected with pigmented villonodular knee synovitis and knee meniscal tears, or persistent gonoarthitis. METHODS: Multiple synovial biopsies of the knee were performed by arthroscopy in five consecutive patients affected by diffuse pigmented villonodular knee synovitis and in 12 patients affected by knee meniscal tears (n. 6) or persistent active gonarthritis (n. 6), recruited from the patients attending the Rheumatology Day Surgery Outpatient Clinic of the University of Padova Hospital. The ethics committee approved the study protocol and the participants signed consent statements after being informed about the content of the study. The diagnosis was made on the basis of a histological examination. The colony-stimulating factor-1 gene expression was assessed by reverse transcription followed by real-time polymerase chain reaction. RESULTS: The detection by RT-PCR of synovial colony-stimulating factor-1 mRNA showed a wide spectrum of expression in the three groups of distinct knee joint disease affected patients, with significantly higher level of colony-stimulating factor-1 mRNA expression in synovial tissue of pigmented villonodular synovitis, in comparison to that of knee meniscal injuries and persistent gonoarthritis patients. CONCLUSIONS: Our findings point out to an important role of colony-stimulating factor-1 in pigmented villonodular knee synovitis disease process and support the idea that colony-stimulating factor 1/colony-stimulating factor-1 receptor interaction may represent a potential therapeutic target of this disease. PMID- 21640050 TI - Association between vascular cell adhesion molecule 1 and radiographic hand osteoarthritis. AB - BACKGROUND: A recent study showed that the level of soluble Vascular Cell Adhesion Molecule 1 (VCAM-1) emerged as a strong and independent predictor of the risk of hip and knee joint replacement due to severe osteoarthritis (OA). Therefore, we hypothesized that soluble VCAM-1 level can be associated with prevalence and severity of radiographic hand OA. OBJECTIVES: To evaluate the association between soluble VCAM-1, and radiographic hand OA in a large community based sample. METHODS: The study population comprised Chuvashians (473 males age 45.90+/-17.16; and 451 females, age 46.18+/-16.70 years). OA was evaluated for 14 joints of each hand using Kellgren and Lawrence (K-L). OA was characterised by the number of affected joints and by the presence of at least one affected joint. The VCAM-1 level was determined by a sandwichenzyme immunoassay technique using ELISA-OSTEO kit. Statistical analyses included multiple linear and logistic regressions. RESULTS: The linear regression model showed a significant association of a number of affected joints with VCAM-1 level (p=0.004) after adjustment for age, sex, BMI. In the logistic regression model the level of association between the presence of at least one affected joint and soluble VCAM 1 level was p=0.070 (OR(95%CI): 1.003 (1.000, 1.007)). CONCLUSIONS: In this cross sectional population-based study, we found that the serum level of soluble VCAM-1 level is positively associated with the number of affected joints of hand OA. PMID- 21640051 TI - Obesity, adipose tissue and rheumatoid arthritis: coincidence or more complex relationship? AB - In the last two decades we have witnessed a boost in scientific interest and knowledge of adipose tissue biology to such an extent that it was promoted to an active endocrine organ. Adipose tissue is not just related to body weight and appetite regulation. It is also implicated in obesity, a low-grade inflammatory state, as well as inflammatory conditions including rheumatoid arthritis (RA), an autoimmune disease where anti- and pro-inflammatory cytokine balance is critical. All major adipose derived products, simply termed adipokines, like leptin, adiponectin, visfatin and resistin, reportedly participate in inflammation and immunity. In this review we explore in depth the relationship between adipose tissue and RA, with focus on possible mechanisms, beyond observations about circulating or synovial levels, and special reference to future perspectives and clinical implications. PMID- 21640052 TI - The potential roles for novel biomarkers in rheumatoid arthritis assessment. AB - Comprehensive management of rheumatoid arthritis (RA) requires regular monitoring of disease activity, functional status, and structural damage to facilitate optimal patient outcomes. Tight control strategies have been successfully used in other diseases including diabetes and hypertension. Tight control requires frequent disease activity measurements in order to tailor treatment for individual patients, resulting in improved patient outcomes. Current monitoring measures used in clinical practice are largely driven by subjective evaluation of signs and symptoms, which are critical but limited by assessor variability and may not reflect true biological change in a timely manner. Research suggests that novel biomarkers may provide quantitative, objective assessments of disease activity and structural damage risk in RA, which are not captured by current measures. The simultaneous use of multiple biomarkers in a single test algorithm may provide a more comprehensive quantitative representation of the overall complex heterogeneous biology of RA. This article reviews the current management strategies for monitoring RA and the potential impact that multi-biomarker assays may have on RA assessment, which may further improve clinical outcomes. PMID- 21640053 TI - Erythema nodosum-like skin lesions associated with Behcet's disease show 18F fluorodeoxyglucose uptake on PET/CT. PMID- 21640054 TI - Clinical presentation and salivary gland histopathology of paediatric primary Sjogren's syndrome. AB - OBJECTIVES: Explore the presentation, diagnostic criteria and exocrine gland histopathology of paediatric primary Sjogren's syndrome (PPSjS). METHODS: A case series of 8 children is reported and American-European Consensus Group (AECG 2002) criteria were examined, as well as minor labial salivary and lachrymal gland biopsies, which were scored by a pathologist blinded to outcome. For all cases, connective tissue diseases and parotid-related infectious disease were excluded. RESULTS: Age at onset varied from 5-13 years old; 6 were females, all followed from diagnosis up to the last visit (1-10 years). The main features at presentation were recurrent tender parotid swelling and sialectasis imaging, with decreased salivary function assessed by Tc-99 scintigraphy. Mild sicca symptoms were observed in 4/8 cases. Systemic features, including fatigue, myalgia, arthritis, tenosynovitis, joint contractures, transient Raynaud's and high ESR, were recorded at onset. Autoantibody profile was unremarkable for diagnosis, while lymphocytic infiltration of labial salivary glands and sialectasis were observed in all biopsies (8/8). In lachrymal glands, massive lymphocytic infiltration and lymphocytic gastritis were observed during complementary assessment. Flares were treated with low dose steroids and long-term use of hydroxychloroquine (5/8), although only 3/8 fulfilled AECG-2002 diagnostic criteria, throughout the disease course. CONCLUSIONS: PPSjS is rare, slowly progressive and its early presentation is variable. Standardised diagnostic algorithms should include recurrent parotid swelling and early diagnosis should rely mostly on salivary and lachrymal gland histopathology in this age group. PMID- 21640055 TI - Early determinants of atherosclerosis in paediatric systemic lupus erythematosus. AB - OBJECTIVES: To assess traditional and non-traditional cardiovascular risk factors and to determine the prevalence and correlates of early vascular markers of atherosclerosis in paediatric systemic lupus erythematosus (pSLE). METHODS: Fifty four adolescents with pSLE had cardiovascular risk factor assessment, disease activity and vascular testing including carotid intima-media thickness (CIMT), flow-mediated dilatation (FMD), arterial stiffness measures, and myocardial perfusion studies. RESULTS: The traditional risk factors of hypertension, elevated triglycerides, apolipoprotein B, haemoglobin A1c and insulin levels and non-traditional risk factors of elevated homocysteine and fibrinogen were present (all p<0.001). Some arterial stiffness measures, central pulse wave velocity and characteristic impedance were elevated (p<0.001), but CIMT, FMD and myocardial perfusion were normal. Cumulative prednisone dose correlated with total cholesterol (r=0.5790, p<0.001) and elevated LDL-C (r=0.4488, p=0.0012). Hydroxychloroquine treatment correlated negatively with total cholesterol (r= 0.4867, p=0.0002), LDL-C (r=-0.4805, p=0.0002) and apolipoprotein B (r=-0.4443, p=0.0011). In multivariate analysis LDL-C correlated with cumulative prednisone dose and negatively with hydroxychloroquine treatment (R2=0.40, p<0.001). CONCLUSIONS: An increased burden of traditional and non-traditional risk factors and early evidence of insulin resistance and increased central arterial stiffness were present in paediatric SLE. Disease-specific and therapy-related factors are likely modifying these cardiovascular risk profiles warranting prospective longitudinal studies. PMID- 21640056 TI - Conservative treatment for urinary incontinence in Men After Prostate Surgery (MAPS): two parallel randomised controlled trials. AB - OBJECTIVE: To determine the clinical effectiveness and cost-effectiveness of active conservative treatment, compared with standard management, in regaining urinary continence at 12 months in men with urinary incontinence at 6 weeks after a radical prostatectomy or a transurethral resection of the prostate (TURP). BACKGROUND: Urinary incontinence after radical prostate surgery is common immediately after surgery, although the chance of incontinence is less after TURP than following radical prostatectomy. DESIGN: Two multicentre, UK, parallel randomised controlled trials (RCTs) comparing active conservative treatment [pelvic floor muscle training (PFMT) delivered by a specialist continence physiotherapist or a specialist continence nurse] with standard management in men after radial prostatectomy and TURP. SETTING: Men having prostate surgery were identified in 34 centres across the UK. If they had urinary incontinence, they were invited to enroll in the RCT. PARTICIPANTS: Men with urinary incontinence at 6 weeks after prostate surgery were eligible to be randomised if they consented and were able to comply with the intervention. INTERVENTIONS: Eligible men were randomised to attend four sessions with a therapist over a 3-month period. The therapists provided standardised PFMT and bladder training for male urinary incontinence and erectile dysfunction. The control group continued with standard management. MAIN OUTCOME MEASURES: The primary outcome of clinical effectiveness was urinary incontinence at 12 months after randomisation, and the primary measure of cost-effectiveness was incremental cost per quality-adjusted life-year (QALY). Outcome data were collected by postal questionnaires at 3, 6, 9 and 12 months. RESULTS: Within the radical group (n = 411), 92% of the men in the intervention group attended at least one therapy visit and were more likely than those in the control group to be carrying out any PFMT at 12 months {adjusted risk ratio (RR) 1.30 [95% confidence interval (CI) 1.09 to 1.53]}. The absolute risk difference in urinary incontinence rates at 12 months between the intervention (75.5%) and control (77.4%) groups was -1.9% (95% CI -10% to 6%). NHS costs were higher in the intervention group [L 181.02 (95% CI L 107 to L 255)] but there was no evidence of a difference in societal costs, and QALYs were virtually identical for both groups. Within the TURP group (n = 442), over 85% of men in the intervention group attended at least one therapy visit and were more likely to be carrying out any PFMT at 12 months after randomisation [adjusted RR 3.20 (95% CI 2.37 to 4.32)]. The absolute risk difference in urinary incontinence rates at 12 months between the intervention (64.9%) and control (61.5%) groups for the unadjusted intention-to-treat analysis was 3.4% (95% CI -6% to 13%). NHS costs [L 209 (95% CI L 147 to L 271)] and societal costs [L 420 (95% CI L 54 to L 785)] were statistically significantly higher in the intervention group but QALYs were virtually identical. CONCLUSIONS: The provision of one-to-one conservative physical therapy for men with urinary incontinence after prostate surgery is unlikely to be effective or cost-effective compared with standard care that includes the provision of information about conducting PFMT. Future work should include research into the value of different surgical options in controlling urinary incontinence. PMID- 21640057 TI - Prevalence of dental caries and dental care utilisation in preschool urban children enrolled in a comparative-effectiveness study. AB - AIM: To assess dental caries prevalence and dental care utilisation in pre-school children enrolled in urban childcare centres that participated in a comparative effectiveness study. STUDY DESIGN: Cross-sectional study. METHODS: Caries prevalence was determined in a cohort of children 12-60 months of age. Eligible children were randomised into two groups: group one received a traditional visual/tactile oral examination and group two received a teledentistry examination. Questionnaires were administered to the children's parents/guardians to gather demographics and information about using dental and medical services. RESULTS: Of 234 children examined, approximately 28% had caries experience. The mean dfs score was 1.56 with a range of 0-34 carious surfaces. The mean dfs score for the children examined by means of teledentistry was 1.75 and for the children examined by means of the traditional visual/tactile method mean dfs was 1.40; the means between the two groups were not significantly different. Twenty-six children showed evidence of being treated for dental caries. According to the parents, 31.5% of the children had never had a dental check-up before, only 3% of the children were lacking dental insurance and majority of the parents (92%) did not perceive accessing dental care for the children as a problem. STATISTICS: The Wilcoxon Mann-Whitney test and the Kruskal-Wallis test were used to assess statistical differences among groups of children. CONCLUSIONS: The data showed that 28% of the children had caries and, of these, 61% had never been treated for caries, thus indicating that continued efforts are needed to improve oral health care utilisation by inner-city preschool children. PMID- 21640058 TI - Socio-behavioural factors influencing oral health of 12 and 15 year old Greek adolescents. A national pathfinder survey. AB - AIM: To estimate the frequency in use of oral health services, oral health self assessment, oral hygiene practices of 12- and 15-year-old Greek children and adolescents, to investigate possible influences of these factors and other socio demographic parameters on oral health. METHODS: A stratified cluster sample of 1224 12 year old and 1257 15 year old children and adolescents of Greek nationality were selected and examined by calibrated examiners. Clinical examination included recording of caries according to the British association for the study of community dentistry diagnostic criteria, while oral hygiene and periodontal status were assessed by means of the simplified debris and the community periodontal index. Subjects were interviewed to answer questions on socio-behavioural risk factors through a structured questionnaire. The student's t-test and ANOVA were used for statistical evaluation of the means and the chi square test was applied for statistical comparison of the proportions. Multiple linear regression analysis was performed to assess the statistical association between variables. RESULTS: Caries experience and untreated caries were significantly higher among children and adolescents who visited the dentist only when in pain or for restoring a tooth, compared to those visiting for check-ups or prevention and having more frequent application of topical fluorides. Tooth brushing, at least twice a day, and flossing were significantly associated with periodontal and oral hygiene status, but not with caries presence. The multivariate analysis revealed that parental educational status and reason for visiting a dentist were strong determinants for caries experience and oral hygiene status but not for periodontal health of children and adolescents. CONCLUSIONS: This study has identified several socio-demographic and behavioural determinants for dental caries, oral hygiene and periodontal health of Greek children and adolescents. PMID- 21640059 TI - Oral health related quality of life of Greek adolescents: a cross-sectional study. AB - AIM: To investigate the impact of oral health status on the quality of life of a cross-section of adolescents belonging to different population groups in different regions of Greece, using the oral health impact profile-short form (OHIP-14), one of the most widely known instruments used for the measurement of disability and discomfort due to oral conditions. METHODS: A random sample consisting of a total of 515 Greek adolescents between the ages of 15-18 years (mean 16.1+/-0.9) were selected from different urban and rural areas according to the last census. A self-administrated questionnaire was designed including the OHIP-14 validated for the Greek language, and face to- face interviews were conducted by one dentist trained in oral health related quality of life (OHRQoL) terms. Associations of the total OHIP-14 score and its seven subscales along with the self-perceived quality of life were evaluated with Spearman correlations. RESULTS: Internal reliability returned a very good internal consistency with a Cronbach alpha of 0.86. The subjects had an overall weighted OHIP-14 score of 1.24 (SD 2.04) meaning that there was an impact of oral health on the overall quality of life. Five of the seven subscales of the OHIP-14 tool were found to have significant correlations for the inhabitants of the different areas. Specifically, important and significant correlations were discovered for functional limitation (p<0.01), handicap (p<0.05) and social disability (p<0.01) both for the metropolitan/non-metropolitan as well as the urban rural distinction. No correlations were found between the OHIP-14 scores, or of any of its sub-scales, with the parental education level and occupation. When self assessed oral and general health statuses were considered to be 'bad' the OHIP-14 returned increased scores. CONCLUSIONS: Dental and oral health conditions are factors that do impact on the quality of life of adolescents. PMID- 21640060 TI - Oral health in young individuals with foreign and Swedish backgrounds--a ten-year perspective. AB - AIM: To investigate oral health status and coherent determinants in children with foreign backgrounds compared with children with a Swedish background, during a ten year period. DESIGN AND METHODS: In 1993 and 2003, cross-sectional studies with random samples of individuals in the age groups 3, 5, 10 and 15 years were performed in Jonkoping, Sweden. All the individuals were personally invited to a clinical and radiographic examination of their oral health status. They were also asked about their attitudes to and knowledge of teeth and oral health care habits. The final study sample comprised 739 children and adolescents, 154 with a foreign background (F cohort) and 585 with a Swedish background (S cohort). RESULTS: In both 1993 and 2003, more 3- and 5 year olds in the S cohort were caries-free compared with the F cohort. In 1993, dfs was higher among 3- and 5 year olds in the F cohort (p<0.01) compared with the S cohort. In 2003, dfs/DFS was statistically significantly higher in all age groups among children and adolescents in the F cohort compared with the S cohort. When it came to proximal tooth surfaces, the percentages of individuals who were caries-free, with initial carious lesions, with manifest carious lesions and with restorations among 10 year-olds in the F cohort were 55%, 23%, 4% and 18% in 1993. The corresponding figures for the S cohort were 69%, 20%, 6% and 5% respectively. In 2003, the values for the F cohort were 54%, 29%, 4% and 13% compared with 82%, 12%, 1% and 5% in the S cohort. In 2003, the odds of being exposed to dental caries among 10- and 15-yearolds in the F cohort, adjusted for gender and age, were more than six times higher (OR=6.3, 95% CI:2.51-15.61; p<0.001) compared with the S cohort. CONCLUSIONS: There has been a decline in caries prevalence between 1993 and 2003 in all age groups apart from 3-year-olds. However, the improvement in dfs/DFS was greater in the S cohort compared with the F cohort in all age groups. The difference between the F and S cohorts in terms of dfs/ DFS was larger in 2003 compared with 10 years earlier. In 2003, the odds ratio for being exposed to dental caries was almost six times higher for 10- and 15-year-olds with two foreign-born parents compared with their Swedish counterparts. PMID- 21640061 TI - Use of the 'Hall technique' for management of carious primary molars among Scottish general dental practitioners. AB - AIM: To assess the current awareness, usage and opinion of the Hall technique as a restorative option for primary molars in Scottish general dental practice; and to identify preferences for methods of further training, if desired, for those not currently using the technique. STUDY DESIGN: A postal questionnaire was sent to a random sample of Scottish general dental practitioners (GDPs) (n= 1207). Half of all GDPs within each health board were mailed. All analyses have been carried out in Minitab (version 15). The study is primarily descriptive and uses frequency distributions and cross-tabulations. Percentages are reported with p5% confidence intervals. Characteristics of the whole sample were reported. However when reporting the use of the Hall technique, only those GDP's reporting to treat children, at least sometimes are considered. RESULTS: Following two mail-shots, the overall response rate was 59% (715/1207). Eighty-six percent (616/715) of respondents were aware of the Hall technique as a method of restoring primary molars and 48 % (n=318) were currently using the Hall technique. Of those GDPs who never used the Hall technique (51% of total respondents; n=340), 46% (n=157) indicated they were either 'very interested' or 'interested' in adopting the Hall technique into their clinical practice. The preferred source for further training was via a section 63 continuing professional development (CPD) course, incorporating a practical element. CONCLUSIONS: Of those GDPs in Scotland who responded to the questionnaire, an unexpectedly high number were already using the Hall technique in their practice, and among those not currently using it, there is a demand for training. PMID- 21640062 TI - The effect of fluoride slow-releasing devices on fluoride in plaque biofilms and saliva: a randomised controlled trial. AB - AIM: To investigate the effect of the fluoride sustained slow-releasing device (FSSRD) on F levels in unstimulated saliva and undisturbed plaque biofilms over 7 days. The investigation also aimed at studying the effect of patient age on fluoride (F) concentrations in dental plaque biofilms and unstimulated saliva while using the FSSRD. STUDY DESIGN: This was a randomised control double blind cross-over study. METHODS: Sixty-five participants between the ages of 6-35 years took part in this study. The mean age of all the participants was 19.22 years with 24 participants aged between 6-16 years (mean age=11.8 years) and 41 participants aged between 16-35 years (mean age=23.57 years). Plaque biofilms were collected using a modification of the plaque generating device (MPGD) [Robinson et al., 1997]. During the whole study period, including a 7 day washout period before the first leg, all participants were asked to use only non fluoridated toothpaste and to avoid where possible high fluoride containing foods or drinks. Whenever possible, the FSSRD/placebo device (PD) was attached to the second permanent molar, while the MPGD was attached to the first permanent molars in the same upper dental quadrant. At the end of each leg of the study whole, mixed unstimulated saliva was collected from all participants and analysed for F concentration. STATISTICS: Statistical analysis using paired sample t-test was used to compare the results of F level between test and control groups, while, Spearman's correlation coefficient was used to test the relationship between patient's age and plaque weight against F concentration in plaque and saliva. RESULTS: There was no statistically significant difference in F concentration in plaque and saliva between the test and control groups when analysed using paired sample t-test (p>0.05). There was no correlation between patient's age and F concentration in dental plaque and saliva (p>0.05), in both the test and control groups. CONCLUSIONS: Our data showed no effect of the FSSRD in raising F concentrations in dental plaque and unstimulated whole saliva after 7 days. Such levels may require longer periods to become established. PMID- 21640063 TI - Disease outcome for children who present with oral manifestations of Crohn's disease. AB - AIM: To describe the outcome for children with oral Crohn's disease (OCD) at diagnosis, and to determine if there was a difference in the Paediatric Crohn's Disease Activity Index (PCDAI) scores between those with and those without oral lesions at follow-up. METHODS: Thirty-one patients with OCD who had enrolled in two previous studies were invited to participate. Clinical and laboratory data were collected to calculate the PCDAI. Details of the management of Crohn's disease were also recorded. RESULTS: Twenty-four of 31 patients participated (77%), of whom 17 were boys (M:F = 2.4:1). Mean age at follow-up was 15.7 years (SD 1.98, range 11.9-19.7 years). Mean duration of follow-up was 55 months (SD 22, range 20-97 months). Oral manifestations were present at follow-up in 7 (29%) of 24 patients. There were no differences between patients with and without OCD at follow-up with regard to medical treatments received or intestinal disease location. There was no difference in median PCDAI scores between those who had and those who had not oral lesions at follow-up. CONCLUSIONS: OCD resolved in the majority of children treated for intestinal Crohn's disease. The occurrence of mouth lesions during follow-up of children who had oral manifestations at initial diagnosis was not a marker for Crohn's disease activity elsewhere in the intestinal tract. PMID- 21640064 TI - Evaluation of a resin modified glass ionomer serving both as indirect pulp therapy and as restorative material for primary molars. AB - AIM: The successful performance of resin modified glass ionomer cement (RMGIC) as an indirect pulp therapy (IPT) dressing material and, independently, as a class II restoration material for primary molars has been adequately documented. This study investigates if an RMGIC can be successfully used in a dual capacity. METHODS: A total of 61, 3.5-8.5 year old children with deep proximal carious lesions in one or more primary molars, radiographically suspected of pulp exposure, were included. Teeth exclusion criteria were: continuous unprovoked pain and an exposed or a non-vital pulp. Following adequate caries removal, RMGIC restorations were placed. The teeth were followed up for signs/symptoms and for restoration survival according to modified United States Public Health Service (USPHS) criteria. Descriptive statistics and a Fisher's exact test were applied. RESULTS: The prospective study was completed with 86 restorations in 41 first and 45 second primary molars. Three of the 86 teeth showed pulp necrosis and the remaining 83 (96.5%) were a) asymptomatic for a mean clinical observation time of 31.9 months (SD 10.8) and b) with no radiographic signs of pathology for 25.7 months (SD 10.3). Seventy six of these 83 (91.6%) class II-'extended' restorations were acceptable, exhibiting varying levels of occlusal wear. CONCLUSIONS: A single application of RMGIC was used successfully for over 2 1/2 years as an IPT and restoration material in proximal caries of primary molars. PMID- 21640065 TI - Short communication: Dmfs and BMI in preschool Greek children. An epidemiological study. AB - AIM: To investigate the relationship between dental caries (dmfs) and body mass index (BMI) categories in 3-5.5 year old children in Thessaloniki, Greece. METHODS: The study was conducted in 18 municipal day care centres and involved 361 children. The dmfs was determined on site by one calibrated examiner using disposable dental mirrors and a penlight. The height and weight of the children were measured on site by a nutritionist, who grouped them into four BMI categories, shown in the results. STATISTICS: The estimation of the relationship between the BMI and dmfs values was based on a generalized linear model (Poisson log-linear regression) while the sequential Bonferroni method was used for pair wise comparisons between BMI categories. RESULTS: Mean dmfs values for each BMI category were: 1.02 (SD=2.41) for the underweight (n=44), 0.74 (SD=2.24) for the normal weight (n=281), 1.88 (SD=4.28) for the overweight (n=26) and 0.80 (SD=2.53) for the obese (n=10). Overweight children were found to show statistically significant differences in dmfs values compared with both children of normal weight (p<0.001) and those underweight (p=0.015). CONCLUSION: Overweight Greek pre-school children are at higher risk of dental caries. PMID- 21640066 TI - Survey of Naegleria from Taiwan recreational waters using culture enrichment combined with PCR. AB - Naegleria is a free-living amoeba. Pathogenic Naegleria may pose a health risk to people who come in contact with recreational waters. Here, we used Naegleria culture enrichment with PCR to identify the Naegleria species and investigated the distribution of Naegleria spp. in recreational waters including spring water, stream water and raw domestic water in central and southern Taiwan. In this study, Naegleria spp. were detected in 19 (17.8%) of the water samples. The occurrence of Naegleria in raw domestic water was 28.6%, higher than in stream water (14.7%) and in spring water (6.5%). The most frequently identified species exhibiting the closest phylogenetic relationships to the isolates were N. australiensis (n=4) and N. canariensis (n=4), followed by N. clarki (n=3) and N. philippinensis (n=3); N. americana (n=2). N. lovaniensis, N. dobsoni, and N. gruberi were each detected once. The pathogenic species N. fowleri was not detected, probably due to the low incubation temperature; however, the isolates exhibiting the closest phylogenetic relationships to the pathogenic species in mice of PAM, N. australiensis and N. philippinensis, were found. Results of this survey suggest the distribution of Naegleria spp. excluding N. fowleri in recreational waters. It should be considered a potential threat for health associated with human activities in recreational waters. PMID- 21640067 TI - An automated point-of-care system for immunodetection of staphylococcal enterotoxin B. AB - An automated point-of-care (POC) immunodetection system for immunological detection of staphylococcal enterotoxin B (SEB) was designed, fabricated, and tested. The system combines several elements: (i) enzyme-linked immunosorbent assay-lab-on-a-chip (ELISA-LOC) with fluidics, (ii) a charge-coupled device (CCD) camera detector, (iii) pumps and valves for fluid delivery to the ELISA-LOC, (iv) a computer interface board, and (v) a computer for controlling the fluidics, logging, and data analysis of the CCD data. The ELISA-LOC integrates a simple microfluidic system into a miniature 96-well sample plate, allowing the user to carry out immunological assays without a laboratory. The analyte is measured in a sandwich ELISA assay format combined with a sensitive electrochemiluminescence (ECL) detection method. Using the POC system, SEB, a major foodborne toxin, was detected at concentrations as low as 0.1 ng/ml. This is similar to the reported sensitivity of conventional ELISA. The open platform with simple modular fluid delivery automation design described here is interchangeable between detection systems, and because of its versatility it can also be used to automate many other LOC systems, simplifying LOC development. This new POC system is useful for carrying out various immunological and other complex medical assays without a laboratory and can easily be adapted for high-throughput biological screening in remote and resource-poor areas. PMID- 21640068 TI - Moderate heat stress induces state transitions in Arabidopsis thaliana. AB - The effect of temperature on the photosynthetic machinery is crucial for the fundamental understanding of plant physiology and the bioengineering of heat tolerant varieties. In our study, Arabidopsis thaliana was exposed to mild (40 degrees C), short-term heat stress in the dark to evaluate the heat-triggered phosphorylation and migration of light harvesting complex (LHC) II in both wild type (wt) and mutant lacking STN7 kinase. The 77K emission spectra revealed an increase in PSI relative to PSII emission similar to increases observed in light induced state I to state II transitions in wt but not in stn7 mutant. Immunoblotting results indicated that the major LHCII was phosphorylated at threonine sites under heat stress in wt plants but not in the mutant. These results support the proposition that mild heat stress triggers state transitions in the dark similar to light-induced state transitions, which involve phosphorylation of LHCII by STN7 kinase. Pre-treatment of Arabidopsis leaves with inhibitor DBMIB, altered the extent of LHCII phosphorylation and PSI fluorescence emission suggests that activation of STN7 kinase may be dependent on Cyt b(6)/f under elevated temperatures in dark. Furthermore, fast Chl a transient of temperature-exposed leaves of wt showed a decrease in the F(v)/F(m) ratio due to both an increase in F(o) and a decrease in F(m). In summary, our findings indicate that a mild heat treatment (40 degrees C) induces state transitions in the dark resulting in the migration of phosphorylated LHCII from the grana to the stroma region. PMID- 21640070 TI - NMR structural study of the intracellular loop 3 of the serotonin 5-HT(1A) receptor and its interaction with calmodulin. AB - The serotonin (5-HT(1A)) receptor, a G-protein-coupled receptor (GPCR), plays important roles in serotonergic signaling in the central nervous system. The third intracellular loop (ICL3) of the 5-HT(1A) receptor has been shown to be important for the regulation of this receptor through interactions with proteins such as G-proteins and calmodulin. In this study, the ICL3 of 5-HT(1A) receptor was expressed in E. coli and purified. Gel filtration and mass spectrometry were used to confirm the molecular weight of the purified ICL3. Secondary structure analysis using circular dichroism (CD) demonstrated the presence of alpha-helical structures. Backbone assignment of ICL3 was achieved using three-dimensional experiments. A chemical shift index and Talos+ analysis showed that residues E326 to R339 form alpha-helical structure. Residues G256 to S269 of ICL3 were shown to be a novel region that has a molecular interaction with calmodulin in titration assays. Peptide derived from the ICL3 containing residues from G256 to S269 also showed molecular interaction with calmodulin. PMID- 21640069 TI - Synthesis and characterization of degradable multivalent cationic lipids with disulfide-bond spacers for gene delivery. AB - Gene therapy provides powerful new approaches to curing a large variety of diseases, which are being explored in ongoing worldwide clinical trials. To overcome the limitations of viral gene delivery systems, synthetic nonviral vectors such as cationic liposomes (CLs) are desirable. However, improvements of their efficiency at reduced toxicity and a better understanding of their mechanism of action are required. We present the efficient synthesis of a series of degradable multivalent cationic lipids (CMVLn, n=2 to 5) containing a disulfide bond spacer between headgroup and lipophilic tails. This spacer is designed to be cleaved in the reducing milieu of the cytoplasm and thus decrease lipid toxicity. Small angle X-ray scattering demonstrates that the initially formed lamellar phase of CMVLn-DNA complexes completely disappears when reducing agents such as DTT or the biologically relevant reducing peptide glutathione are added to mimic the intracellular milieu. The CMVLs (n=3 to 5) exhibit reduced cytotoxicity and transfect mammalian cells with efficiencies comparable to those of highly efficient non-degradable analogs and benchmark commercial reagents such as Lipofectamine 2000. Thus, our results demonstrate that degradable disulfide spacers may be used to reduce the cytotoxicity of synthetic nonviral gene delivery carriers without compromising their transfection efficiency. PMID- 21640071 TI - Influence of the arrangement and secondary structure of melittin peptides on the formation and stability of toroidal pores. AB - Melittin interactions with lipid bilayers and melittin formed pores are extensively studied to understand the mechanism of the toroidal pore formation. Early experimental studies suggested that melittin peptide molecules are anchored by their positively charged residues located next to the C-terminus to only one leaflet of the lipid bilayer (asymmetric arrangement). However, the recent non linear spectroscopic experiment suggests a symmetric arrangement of the peptides with the C-terminus of the peptides anchored to both bilayers. Therefore, we present here a computational study that compares the effect of symmetric and asymmetric arrangements of melittin peptides in the toroidal pore formation. We also investigate the role of the peptide secondary structure during the pore formation. Two sets of the symmetric and asymmetric pores are prepared, one with a helical peptide from the crystal structure and the other set with a less helical peptide. We observe a stable toroidal pore being formed only in the system with a symmetric arrangement of the less helical peptides. Based on the simulation results we propose that the symmetric arrangement of the peptides might be more favorable than the asymmetric arrangement, and that the helical secondary structure is not a prerequisite for the formation of the toroidal pore. PMID- 21640072 TI - Multivalent protein binding in carbohydrate-functionalized monolayers through protein-directed rearrangement and reorientation of glycolipids at the air-water interface. AB - Multivalent protein binding plays an important role not only in biological recognition but also in biosensor preparation. Infrared reflection absorption spectroscopy and surface plasmon resonance techniques have been used to investigate concanavalin A (Con A) binding to binary monolayers composed of 1,2 di-O-hexadecyl-sn-glycerol and derived glycolipids with the mannose moieties. The glycolipids in the binary monolayers at the air-water interface underwent both lateral rearrangement and molecular reorientation directed by Con A in the subphase favorable to access of the carbohydrate ligands to protein binding pockets for the formation of multivalent binding sites and the minimization of steric crowding of neighboring ligands for enhanced binding. The amounts of specifically bound proteins in the binary monolayers at the air-water interface were accordingly increased in comparison with those in the initially immobilized monolayers at the air-water interface. The directed rearranged binary monolayers with multivalent protein binding were preserved for the preparation of biosensors. PMID- 21640073 TI - Folding studies of purified LamB protein, the maltoporin from the Escherichia coli outer membrane: trimer dissociation can be separated from unfolding. AB - The folding mechanisms for beta-barrel membrane proteins present unique challenges because acquisition of both secondary and tertiary structure is coupled with insertion into the bilayer. For the porins in Escherichia coli outer membrane, the assembly pathway also includes association into homotrimers. We study the folding pathway for purified LamB protein in detergent and observe extreme hysteresis in unfolding and refolding, as indicated by the shift in intrinsic fluorescence. The strong hysteresis is not seen in unfolding and refolding a mutant LamB protein lacking the disulfide bond, as it unfolds at much lower denaturant concentrations than wild type LamB protein. The disulfide bond is proposed to stabilize the structure of LamB protein by clasping together the two sides of Loop 1 as it lines the inner cavity of the barrel. In addition we find that low pH promotes dissociation of the LamB trimer to folded monomers, which run at about one third the size of the native trimer during SDS PAGE and are much more resistant to trypsin than the unfolded protein. We postulate the loss at low pH of two salt bridges between Loop 2 of the neighboring subunit and the inner wall of the monomer barrel destabilizes the quaternary structure. PMID- 21640074 TI - Hypoxia induces beta-amyloid in association with death of RGC-5 cells in culture. AB - Beta-amyloid (Abeta) derived from amyloid precursor protein (APP) has been associated with retinal degeneration in Alzheimer's disease (AD) and glaucoma. This study examined whether hypoxia exposure induces Abeta accumulation in RGC-5 cells. While levels of APP mRNA and protein significantly increased in the cells, elevated abundance of Abeta was also observed in cells and culture medium between 12 or 24 and 48h after 5% O(2) hypoxia treatment. Additionally, there is a close relationship between induction of APP and Abeta and intracellular accumulation of ROS along with loss of mitochondrial membrane potential followed by the death of RGC-5 cells in culture under hypoxia. These results suggest a possible involvement of APP and Abeta in the death of RGCs challenged by hypoxia. PMID- 21640075 TI - Renin-angiotensin system inhibitors suppress azoxymethane-induced colonic preneoplastic lesions in C57BL/KsJ-db/db obese mice. AB - Obesity-related metabolic abnormalities, including chronic inflammation and oxidative stress, increase the risk of colorectal cancer. Dysregulation of the renin-angiotensin system (RAS) also plays a critical role in obesity-related metabolic disorders and in several types of carcinogenesis. In the present study, we examined the effects of an angiotensin-converting enzyme (ACE) inhibitor and angiotensin-II type 1 receptor blocker (ARB), both of which inhibit the RAS, on the development of azoxymethane (AOM)-initiated colonic premalignant lesions in C57BL/KsJ-db/db (db/db) obese mice. Male db/db mice were given 4 weekly subcutaneous injections of AOM (15 mg/kg body weight), and then, they received drinking water containing captopril (ACE inhibitor, 5mg/kg/day) or telmisartan (ARB, 5mg/kg/day) for 7 weeks. At sacrifice, administration of either captopril or telmisartan significantly reduced the total number of colonic premalignant lesions, i.e., aberrant crypt foci and beta-catenin accumulated crypts, compared to that observed in the control group. The expression levels of TNF-alpha mRNA in the colonic mucosa of AOM-treated db/db mice were decreased by captopril and telmisartan. Captopril lowered the expression levels of TNF-alpha, IL-1beta, IL 6, and PAI-1 mRNAs, while telmisartan lowered the expression levels of COX-2, IL 1beta, IL-6, and PAI-1 mRNAs in the white adipose tissues of these mice. In addition, these agents significantly reduced the levels of urinary 8-OHdG, a surrogate marker of oxidative damage to DNA, in the experimental mice. These findings suggested that both ACE inhibitor and ARB suppress chemically-induced colon carcinogenesis by attenuating chronic inflammation and reducing oxidative stress in obese mice. Therefore, targeting dysregulation of the RAS might be an effective strategy for chemoprevention of colorectal carcinogenesis in obese individuals. PMID- 21640076 TI - Differential heterocyclic substrate recognition by, and pteridine inhibition of E. coli and human tRNA-guanine transglycosylases. AB - tRNA-guanine transglycosylases (TGTs) are responsible for incorporating 7 deazaguanine-modified bases into certain tRNAs in eubacteria (preQ(1)), eukarya (queuine) and archaea (preQ(0)). In each kingdom, the specific modified base is different. We have found that the eubacterial and eukaryal TGTs have evolved to be quite specific for their cognate heterocyclic base and that Cys145 (Escherichia coli) is important in recognizing the amino methyl side chain of preQ(1) (Chen et al., Nuc. Acids Res. 39 (2011) 2834 [15]). A series of mutants of the E. coli TGT have been constructed to probe the role of three other active site amino acids in the differential recognition of heterocyclic substrates. These mutants have also been used to probe the differential inhibition of E. coli versus human TGTs by pteridines. The results indicate that mutation of these active site amino acids can "open up" the active site, allowing for the binding of competitive pteridine inhibitors. However, even the "best" of these mutants still does not recognize queuine at concentrations up to 50MUM, suggesting that other changes are necessary to adapt the eubacterial TGT to incorporate queuine into RNA. The pteridine inhibition results are consistent with an earlier hypothesis that pteridines may regulate eukaryal TGT activity (Jacobson et al., Nuc. Acids Res. 9 (1981) 2351 [8]). PMID- 21640077 TI - Role of Pin1 in UVA-induced cell proliferation and malignant transformation in epidermal cells. AB - Ultraviolet A (UVA) radiation (lambda = 320-400 nm) is considered a major cause of human skin cancer. Pin1, a peptidyl prolyl isomerase, is overexpressed in most types of cancer tissues and plays an important role in cell proliferation and transformation. Here, we demonstrated that Pin1 expression was enhanced by low energy UVA (300-900 mJ/cm(2)) irradiation in both skin tissues of hairless mice and JB6 C141 epidermal cells. Exposure of epidermal cells to UVA radiation increased cell proliferation and cyclin D1 expression, and these changes were blocked by Pin1 inhibition. UVA irradiation also increased activator protein-1 (AP-1) minimal reporter activity and nuclear levels of c-Jun, but not c-Fos, in a Pin1-dependent manner. The increases in Pin1 expression and in AP-1 reporter activity in response to UVA were abolished by N-acetylcysteine (NAC) treatment. Finally, we found that pre-exposure of JB6 C141 cells to UVA potentiated EGF inducible, anchorage-independent growth, and this effect was significantly suppressed by Pin1inhibition or by NAC. PMID- 21640078 TI - Mechanical stretch of sympathetic neurons induces VEGF expression via a NGF and CNTF signaling pathway. AB - Mechanical stretch has been shown to increase vascular endothelial growth factor (VEGF) expression in cultured myocytes. Sympathetic neurons (SN) also possess the ability to express and secrete VEGF, which is mediated by the NGF/TrkA signaling pathway. Recently, we demonstrated that SN respond to stretch with an upregulation of nerve growth factor (NGF) and ciliary neurotrophic factor (CNTF). Whether stretch increases neuronal VEGF expression still remains to be clarified. Therefore, SN from the superior cervical ganglia of neonatal Sprangue Dawley rats were exposed to a gradual increase of stretch from 3% up to 13% within 3days (3%, 7% and 13%). Under these conditions, the expression and secretion of VEGF was analyzed. Mechanical stretch significantly increased VEGF mRNA and protein expression (mRNA: control=1 vs. stretch=3.1; n=3/protein: control=1 vs. stretch=2.7; n=3). ELISA experiments to asses VEGF content in the cell culture supernatant showed a time and dose dependency in VEGF increment due to stretch. NGF and CNTF neutralization decreased stretch-induced VEGF augmentation in a significant manner. This response was mediated in part by TrkA receptor activation. The stretch-induced VEGF upregulation was accompanied by an increase in HIF-1alpha expression. KDR levels remained unchanged under conditions of stretch, but showed a significant increase due to NGF neutralization. In summary, SN respond to stretch with an upregulation of VEGF, which is mediated by the NGF/CNTF and TrkA signaling pathway paralleled by HIF-1alpha expression. NGF signaling seems to play an important role in regulating neuronal KDR expression. PMID- 21640079 TI - The distribution of mannose-6-phosphate receptors changes from newborns to adults in rat liver. AB - The co-existence of two types of mannose-6-phosphate receptors (CD-MPR and CI MPR) in most cell types is still not well explained. Some evidence suggests that the CI-MPR could be actively involved in the regulation of growth factors in the early stages of mammalian organ development. In this study, it was demonstrated that both receptors are distributed in a non-overlapping fashion in rat liver, and that the distribution of CI-MPR changes over a percoll gradient between newborn and adult animals. By using marker proteins it was observed that in newborns the CI-MPR is located both in intracellular fractions and in fractions that coincide with a plasma membrane marker, whereas in adults it is only detected in intracellular fractions. It was also noted that N-acetyl-beta-D glucosaminidase distribution is closer to CI-MPR than to CD-MPR and that acid phosphatase did not match with any receptor. This evidence may also suggest that both receptors have different functions, mainly at early stages in the development of organs. PMID- 21640080 TI - Role of a tyrosine phosphorylation of SMG-9 in binding of SMG-9 to IQGAP and the NMD complex. AB - SMG-9 is a component of the NMD complex, a heterotetramer that also includes SMG 1 and SMG-8 in the complex. SMG-9 was also originally identified as a tyrosine phosphorylated protein but the role of the phosphorylation is not yet known. In this study, we determined that IQGAP protein, an actin cytoskeleton modifier acts as a binding partner with SMG-9 and this binding is regulated by phosphorylation of SMG-9 at Tyr-41. SMG-9 is co-localized with IQGAP1 as a part of the process of actin enrichment in non-stimulated cells, but not in the EGF-stimulated cells. Furthermore, an increase in the ability of SMG-9 to bind to SMG-8 occurs in response to EGF stimulation. These results suggest that tyrosine phosphorylation of SMG-9 may play a role in the formation of the NMD complex in the cells stimulated by the growth factor. PMID- 21640081 TI - Decreased sarcolipin protein expression and enhanced sarco(endo)plasmic reticulum Ca2+ uptake in human atrial fibrillation. AB - Sarcolipin (SLN), a key regulator of cardiac sarco(endo)plasmic reticulum (SR) Ca(2+) ATPase, is predominantly expressed in atria and mediates beta-adrenergic responses. Studies have shown that SLN mRNA expression is decreased in human chronic atrial fibrillation (AF) and in aortic banded mouse atria; however, SLN protein expression in human atrial pathology and its role in atrial SR Ca(2+) uptake are not yet elucidated. In the present study, we determined the expression of major SR Ca(2+) handling proteins in atria of human AF patients and in human and in a mouse model of heart failure (HF). We found that the expression of SR Ca(2+) uptake and Ca(2+) release channel proteins are significantly decreased in atria but not in the ventricles of pressure-overload induced HF in mice. In human AF and HF, the expression of SLN protein was significantly decreased; whereas the expressions of other major SR Ca(2+) handling proteins were not altered. Further, we found that the SR Ca(2+) uptake was significantly increased in human AF. The selective downregulation of SLN and enhanced SR Ca(2+) uptake in human AF suggest that SLN downregulation could play an important role in abnormal intracellular Ca(2+) cycling in atrial pathology. PMID- 21640082 TI - Identification of a novel CaMKK substrate. AB - Ca(2+)/calmodulin-dependent protein kinase kinase (CaMKK) phosphorylates and activates specific downstream protein kinases including CaMKI, CaMKIV and 5'-AMP activated protein kinase. In order to examine the variety of CaMKK-mediated signaling pathways, we searched for novel CaMKK substrate(s) using N(6)-(1 methylbutyl)-ATP and genetically engineered CaMKKalpha mutant, CaMKKalpha (Phe(230)Gly), that was capable of utilizing this ATP analogue as a phosphate donor. Incubation of rat brain extracts with recombinant CaMKKalpha (Phe(230)Gly), but not with wild-type kinase, in the presence of N(6)-(1 methylbutyl)-ATP and Ca(2+)/CaM, induced significant threonine phosphorylation of a 50kDa protein as well as CaMKI phosphorylation at Thr(177). The 50kDa CaMKK substrate was partially purified by using serial column chromatography, and was identified as Syndapin I by LC-MS/MS analysis. We confirmed that recombinant Syndapin I was phosphorylated by CaMKKalpha and beta isoforms at Thr(355)in vitro. Phosphorylation of HA-Syndapin I at Thr(355) in transfected HeLa cells was significantly induced by co-expression of constitutively active mutants of CaMKK isoforms. This is the first report that CaMKK is capable of phosphorylating a non kinase substrate suggesting the possibility of CaMKK-mediated novel Ca(2+) signaling pathways that are independent of downstream protein kinases. PMID- 21640083 TI - Galectin-3 secretion and tyrosine phosphorylation is dependent on the calpain small subunit, Calpain 4. AB - Cell adhesion and migration are important events that occur during embryonic development, immune surveillance, wound healing and in tumor metastasis. It is a multi-step process that involves both mechanical and biochemical signaling that results in cell protrusion, adhesion, contraction and retraction. Each of these events generates mechanical forces into the environment measured as traction forces. We have previously found that the calpain small subunit, Calpain 4, is required for normal traction forces, and that this mechanism is independent of the catalytic activities of the holoenzymes that are formed between Calpain 4 and each of the proteolytic heavy chains of Calpain 1 and 2. To define a potential mechanism for the Calpain 4 regulation of traction force, we have evaluated the levels of tyrosine phosphorylation, a hallmark of force dependent signaling within focal adhesions. Using 2D gel electrophoresis we compared tyrosine phosphorylation profiles of Calpain 4 deficient mouse embryonic fibroblasts (MEFs) to the levels in wildtype MEFs and MEF's deficient in the large catalytic subunits, Capn1 and Capn2. Of particular interest, was the identification of Galectin-3, a galactose binding protein known to interact with integrins. Galectin-3 has previously been shown to regulate cell adhesion and migration in both normal and tumor cells; however its full mechanism remains elusive. We have found that Calpain 4 is essential for the tyrosine phosphorylation of galectin-3, and its ultimate secretion from the cell, and speculate that its secretion interferes with the production of traction forces. PMID- 21640084 TI - Skp1 stabilizes the conformation of F-box proteins. AB - The SCF ubiquitin ligase complex consists of four components, Skp1, Cul1, ROC1/Rbx1, and a variable subunit F-box protein, which serves as a receptor for target proteins. The F-box proteins consist of an N-terminal ~40 amino acid F-box domain that binds to Skp1 and the C-terminal substrate-binding domain. We have reported previously that Fbs1 and Fbs2 are N-linked glycoprotein-specific F-box proteins. In addition, other three F-box proteins, Fbg3, Fbg4, and Fbg5, show high homology to Fbs1 and Fbs2, but their functions remain largely unknown. Here we report that Skp1 assists in correct folding of exogenously expressed F-box proteins. Fbs2 as well as Fbg3, Fbg4, and Fbg5 proteins formed SCF complexes but did not bind to N-glycoproteins when exogenously expressed alone. However, co expression of Fbs2 and Fbg5 with Skp1 facilitated their binding to glycoproteins that reacted with ConA. Furthermore, Skp1 increased the cellular concentrations of F-box proteins by preventing aggregate formation. These observations suggest that Skp1 plays an important role in stabilizing the conformation of these F-box proteins, which increases their expression levels and substrate-binding. PMID- 21640085 TI - Effect of thymol on peripheral blood mononuclear cell PBMC and acute promyelotic cancer cell line HL-60. AB - Thymol, a naturally occurring phenolic compound, has been known for its antioxidant, anti microbial, and anti inflammatory activity. Thymol has also been reported as anti-cancer agent, but its anti-cancer mechanism has not yet been fully elucidated. Thus, we aimed to investigate anticancer activity of thymol on HL-60 (acute promyelotic leukemia) cells. In our study, thymol demonstrated dose dependent cytotoxic effects on HL-60 cells after 24h of exposure. However, thymol did not show any cytotoxic effect in normal human PBMC. The cytotoxic effect of thymol on HL-60 cells appears to be associated with induction of cell cycle arrest at sub G0/G1 phase, and apoptotic cell death based on genomic DNA fragmentation pattern. Thymol also showed significant increase in production of reactive oxygen species (ROS) activity, increase in mitochondrial H(2)O(2) production and depolarization of mitochondrial membrane potential. On performing Western Blot analysis, thymol showed increase in Bax protein level with a concomitant decrease in Bcl2 protein expression in a dose dependent manner. Our study also showed activation of caspase -9, -8 and -3 and concomitant PARP cleavage, which is the hallmark of caspase-dependent apoptosis. Moreover, to rule out the involvement of other mechanisms in apoptosis induction by thymol, we also studied its effect on apoptosis inducing factor (AIF). Thymol induced AIF translocation from mitochondria to cytosol and to nucleus, thus indicating its ability to induce caspase independent apoptosis. We conclude that, thymol-induced apoptosis in HL-60 cells involves both caspase dependent and caspase independent pathways. PMID- 21640086 TI - P2RX7 genotype association in severe sepsis identified by a novel Multi Individual Array for rapid screening and replication of risk SNPs. AB - BACKGROUND: Functional single nucleotide polymorphisms (SNPs) are relevant to individual therapeutic approaches and may play a role in disease susceptibility. Genome-wide scans, which are now widely applied to detect disease-associated SNPs, provide only limited evidence about SNP associations. Their usefulness as disease markers requires appropriate phenotype analysis and retesting of the gene providing SNP information. Larger data sets of thousands of samples are necessary to confirm the suggested SNPs. METHODS: We applied a newly established microarray based technology that significantly accelerates and simplifies such studies. A tailor-made microarray surface chemistry, sample/probe immobilization and a primer extension reaction are central to the multi-individual array (MIA) platform, which simultaneously identifies the same variable nucleotide in thousands of samples. The set of SNPs to be typed for is highly flexible and can be adapted to the demands of defined clinical questions. RESULTS: A MIA-SNP analysis of functional SNPs in the P2RX7 calcium channel is presented. One risk genotype has been verified by functional analysis using patch clamping. Two clinically relevant genotypes composed of 5 functional SNPs in the P2RX7 gene have been identified in patients with severe sepsis and septic shock, whereas no significant association has been found in patient suffering from hemophagocytic syndromes. CONCLUSIONS: These results support a functional SNP genotyping of the P2RX7 gene in patients at risk of severe sepsis following surgical trauma. PMID- 21640087 TI - New screening tests enrich anti-transglutaminase results and support a highly sensitive two-test based strategy for celiac disease diagnosis. AB - BACKGROUND: The identification of specific serological algorithms allowing the diagnosis of celiac disease (CD) is a new challenge for both the clinic and the laboratory. We compared the diagnostic accuracy of three new tests proposed for CD screening with that of the well established IgA tTG, and ascertained whether any combination of these tools might enhance accuracy in diagnosing CD. METHODS: In sera from 329 CD and 374 control children, the following were assayed: IgA tTG; IgA/IgG, which identify tTG-gliadin complexes (Aeskulisa Celi Check and CeliCheck IgGA); IgA/IgG, which identify deamidated gliadin peptides and tTG (QUANTA Lite(TM) h-tTG/DGP Screen). RESULTS: When specificity was set at 100%, the most sensitive index of CD was IgA tTG (75.7%, cut-off=100U), followed by QUANTA Lite(TM) h-tTG/DGP Screen (65.3%, cut-off 145U), Aeskulisa Celi Check (62.6%, cut-off 909U/mL) and CeliCheck IgGA (59.6%, cut-off 977U/mL). Three algorithms were obtained by combining IgA tTG with each of the new tests. The algorithm obtained by measuring IgA tTG and QUANTA Lite(TM) h-tTG/DGP Screen allowed the correct identification of CD in 78.7% of cases (negative predictive value=97.3%). CONCLUSIONS: The two-test based strategy could be used for the cost effective diagnosis of CD. PMID- 21640088 TI - Pediatric reference intervals for serum alpha-fetoprotein. PMID- 21640089 TI - Biomarkers in Serbian patients with Gaucher disease. AB - OBJECTIVES: The aim of the study was to evaluate the efficiency of the biomarkers chitotriosidase (Chito), total acid phosphatase (TACP), angiotensin converting enzyme (ACE) and ferritin in the diagnosis of Gaucher disease (GD) and to assess the utility of biomarkers for monitoring the effects of enzyme replacement therapy (ERT). DESIGN AND METHODS: Forty treatment-naive Gaucher patients were studied. 27/40 GP were put on ERT and monitored every 6 months. RESULTS: The baseline median values of Chito, TACP, ACE and ferritin were highly elevated in GP: 10216 nmol/mL/h, 26.1 U/L, 253 U/L, 515 MUg/L, and 555 MUg/L, respectively. The only significant difference between mild and moderate GP subgroups is observed for Chito activity (p=0.0116). During ERT, Chito showed the steepest decrease in regard to TACP and ACE, mainly within the first year (71.4%). CONCLUSIONS: Among these biomarkers, Chito proved to be the most useful biomarker for diagnosing GD and monitoring the ERT. PMID- 21640090 TI - Method for monitoring of the protein amino group changes during carbonylation. AB - OBJECTIVES: Carbonylation of the protein amino, guanidino and thiol groups is one of the important causes of vascular complications in diabetes. We developed a simple spectrophotometric method for monitoring of the changes in the protein amino group contents during carbonylation. DESIGN AND METHODS: The method is based on the reaction of amino group with p-benzoquinone in the slightly alkaline media. It was applied during carbonylation in vitro with methylglyoxal and in vivo in 13 patients with type 2 diabetes and 20 healthy persons. RESULTS: The method is simple, fast, precise (RSD in the range of 1.2-1.8%) and accurate (recovery about 100%). The content of amino groups in human serum albumin isolated from diabetics was significantly lower (p<0.01) in comparison with a control group. CONCLUSION: The method developed is suitable for quantification of protein amino groups during in vitro carbonylation as well as for clinical practice. PMID- 21640091 TI - Analysis of serum adenosine deaminase (ADA) and ADA1 and ADA2 isoenzyme activities in HIV positive and HIV-HBV co-infected patients. AB - OBJECTIVES: To determine adenosine deaminase (ADA) activity as a possible diagnostic marker in HIV and HIV-HBV co-infected patients. DESIGN AND METHODS: Blood samples were collected from 72 healthy, 33 HIV positive and 30 HIV-HBV co infected subjects. Blood CD4+ cell count was recorded and serum alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, total ADA, and ADA1 and ADA2 isoenzyme activities were determined. RESULTS: Serum ALT, AST, total ADA and ADA2 isoenzyme activities were significantly higher in HIV positive and HIV-HBV co-infected groups compare to the control (p<0.05), whereas serum ALP showed no differences between groups. CD4+ cell counts markedly decreased in all patients and showed a significant inverse correlation with ADA activities (R(2)=0.589, p<0.001). CONCLUSIONS: Serum ADA was significantly increased in HIV and HIV-HBV co-infections. Therefore, because of its low cost and simplicity to perform, ADA activity might be considered as a useful diagnostic tool among the other markers in these diseases. PMID- 21640092 TI - High sensitivity troponin T concentrations in patients undergoing noncardiac surgery: a prospective cohort study. AB - OBJECTIVES: To determine the proportion of noncardiac surgery patients exceeding the published 99th percentile or change criteria with the high sensitivity Troponin T (hs-TnT) assay. DESIGN AND METHODS: We measured hs-TnT preoperatively and postoperatively on days 1, 2 and 3 in 325 adults. RESULTS: Postoperatively 45% (95% CI: 39-50%) of patients had hs-TnT>=14ng/L and 22% (95% CI:17-26%) had an elevation (>=14ng/L) and change (>85%) in hs-TnT. CONCLUSION: Further research is needed to inform the optimal hs-TnT threshold and change in this setting. PMID- 21640093 TI - Increased serum levels of S100B are related to the severity of cardiac dysfunction, renal insufficiency and major cardiac events in patients with chronic heart failure. AB - OBJECTIVE: To investigate the correlations between S100B and the severity of cardiac dysfunction, renal insufficiency (RI) and prognosis in chronic heart failure (CHF). METHOD: Serum levels of S100B, TNF-alpha, high sensitivity CRP and NT-proBNP were determined in CHF patients with (n=96) and without RI (n=146). Patients with RI only (n=62) and control subjects (n=64) served for comparison. Patients were followed up for one year. RESULTS: S100B levels were higher in CHF patients with a further elevation in those with RI (P<0.01). Serum S100B levels correlated with left ventricular ejection fraction, left ventricular end diastolic volume and NT-proBNP in CHF patients, and eGFR in patients with RI (all P<0.05). Increased S100B levels were associated with major cardiac events (MCE), and were independently associated with the presence of CHF (all P<0.05). CONCLUSION: Increased serum S100B levels were associated with the severity of cardiac dysfunction, RI and an adverse prognosis in CHF patients. It represents an independent risk factor for CHF. PMID- 21640094 TI - Performance evaluation of Elecsys analysis system for anti-cyclic citrullinated peptide detection in comparison with commercially available ELISA assays in rheumatoid arthritis diagnosis. AB - OBJECTIVES: To evaluate performance consistency of electrochemical luminescence with ELISA for anti-cyclic citrullinated peptide (CCP) antibodies detection and define their diagnostic efficacy on rheumatoid arthritis (RA). DESIGN AND METHODS: Serum anti-CCP antibodies of 144 subjects (93 RA, 17 non-RA autoimmune diseases,11 involution diseases and 23 healthy volunteers) were simultaneously determined by Elecsys assay, Immunoscan CCPlus Euro-diagnostica ELISA kit and Euroimmun anti-CCP ELISA kit. RESULTS: The consistent rate and Kappa coefficients among three assays were all more than 0.97. AUCs of ROC for anti-CCP antibodies detection by Elecsys, Euro-diagnostica ELISA and Euroimmun ELISA assays were respectively 0.790, 0.773 and 0.794 (all P values=0.000), and under the optimal cutoff values their diagnostic accurate rates for RA diagnosis were 78.47%, 77.08% and 76.39% respectively. CONCLUSIONS: The diagnostic performance of Elecsys assay for anti-CCP antibodies detection was satisfactory and comparable to that of classical ELISA assay. Under the same specificity Elecsys assay has a slightly but not significantly higher diagnostic accuracy and sensitivity than ELISA assay. PMID- 21640095 TI - Synergistic antihypersensitive effects of pregabalin and tapentadol in a rat model of neuropathic pain. AB - Neuropathic pain is a clinical condition which remains poorly treated and combinations of pregabalin, an antagonist of the alpha2delta-subunit of Ca(2+) channels, with tapentadol, a MU-opioid receptor agonist/noradrenaline reuptake inhibitor, or with classical opioids such as oxycodone and morphine might offer increased therapeutic potential. In the rat spinal nerve ligation model, a dose dependent increase in ipsilateral paw withdrawal thresholds was obtained using an electronic von Frey filament after IV administration of pregabalin (1-10mg/kg), tapentadol (0.316-10mg/kg), morphine (1-4.64 mg/kg) and oxycodone (0.316-3.16 mg/kg), with ED(50) values (maximal efficacy) of 4.21 (67%), 1.65 (94%), 1.70 (96%) and 0.63 mg/kg (100%), respectively. Equianalgesic dose combinations of pregabalin and tapentadol (dose ratio 2.5:1), morphine (2.5:1) or oxycodone (6.5:1) resulted in ED(50) values (maximal efficacy) of 0.83 (89%), 2.33 (97%) and 1.14 mg/kg (100%), respectively. The concept of dose-equivalence suggested an additive interaction of pregabalin and either oxycodone or morphine, while a synergistic interaction was obtained with pregabalin and tapentadol (demonstrated by isobolographic analysis). There was no increase in contralateral paw withdrawal thresholds and no locomotor impairment, as measured in the open field, for the combination of pregabalin and tapentadol; while a significant increase and impairment was demonstrated for the combinations of pregabalin and either morphine or oxycodone. Because combination of pregabalin and tapentadol resulted in a synergistic antihypersensitive activity, it is suggested that, beside the use of either drug alone, this drug combination may offer a beneficial treatment option for neuropathic pain. PMID- 21640096 TI - Resveratrol prevents endothelial nitric oxide synthase uncoupling and attenuates development of hypertension in spontaneously hypertensive rats. AB - Endothelial dysfunction is a hallmark of hypertension and vascular oxidative stress can contribute to endothelial dysfunction and hypertension development. Resveratrol is an antioxidant polyphenol which improves endothelium dependent relaxation, the mechanisms of which are unknown. Also, the role of resveratrol in hypertension remains to be established. The purpose of this study was to investigate the mechanisms of resveratrol induced improvement of endothelial function and establish its role in hypertension. SHR and WKY rats, 3-4 weeks old, were treated with resveratrol in drinking water for 10 weeks, untreated SHR and WKY rats served as controls. At the end of the treatment, control SHR exhibited increased blood pressure, oxidative stress and attenuated endothelium dependent relaxation in comparison to WKY rats. The impaired endothelium function in SHR was associated with lower nitrite/nitrate levels, elevated nitrotyrosine content and eNOS uncoupling. Resveratrol treatment attenuated hypertension development in SHR as indicated by lower blood pressure in resveratrol treated SHR (SHR-R) compared to control SHR. SHR-R also exhibited reduced H(2)O(2) content and elevated superoxide dismutase activity. Resveratrol treatment normalized endothelium dependent vasorelaxation in SHR. In parallel, resveratrol restored nitrite/nitrate levels and normalized nitrotyrosine content in SHR. SHR exhibited increased l-arginine dependent superoxide production which was blocked by NOS inhibitor l-NNA, suggesting eNOS uncoupling. eNOS uncoupling was prevented by resveratrol treatment. In conclusion, early treatment with resveratrol lowers oxidative stress, preserves endothelial function and attenuates development of hypertension in SHR. More importantly, prevention of eNOS uncoupling and NO scavenging could represent novel mechanisms for resveratrol-mediated antihypertensive effects. PMID- 21640097 TI - TRPA1 ion channel in the spinal dorsal horn as a therapeutic target in central pain hypersensitivity and cutaneous neurogenic inflammation. AB - Transient receptor potential ankyrin 1 (TRPA1) is a non-selective, calcium permeable cation channel expressed by a subpopulation of primary afferent nociceptive nerve fibers. On peripheral nerve endings, TRPA1 channel contributes to transduction of chemical and physical stimuli, whereas on the central endings in the spinal dorsal horn, which is the topic of this brief review, it regulates glutamatergic transmission. Blockade of the spinal TRPA1 channel has attenuated mechanical pain hypersensitivity particularly to low-intensity stimulation in various pathophysiological conditions, whereas blockade of the TRPA1 channel mediated regulation of transmission failed to influence baseline pain behavior in healthy control animals. Additionally, blockade of the spinal TRPA1 channel reduced cutaneous neurogenic inflammation, presumably by decreasing drive of spinal interneurons that induce a proinflammatory dorsal root reflex. The spinal TRPA1 channel provides a promising target for development of a selective disease modifying therapy for central pain hypersensitivity. Blockade of the spinal TRPA1 channel-mediated regulation of transmission may also attenuate cutaneous neurogenic inflammation. PMID- 21640098 TI - Enhanced Angiotensin II type 1 receptor expression in leukocytes of patients with chronic kidney disease. AB - Previously, we demonstrated increased Angiotensin II type I receptor expression in leukocytes from patients with untreated, but not in treated, essential hypertension (essential hypertension). We hypothesized that the Angiotensin II AT1 receptor is also increased in leukocytes from patients with chronic kidney disease, however and can still be corrected with combined anti-hypertensive treatment with renin-angiotensin system (RAS) blockers and statins. Blood pressure, cholesterol, renal function oxidative stress parameters, inflammation, and leukocyte Angiotensin II AT1 receptor mRNA expression were measured both on and (6 weeks) off treatment. Data were compared to data of 10 healthy control subjects. Untreated chronic kidney disease patients (n=20) had higher blood pressure, cholesterol and leukocyte Angiotensin II AT1 receptor mRNA expression, but no different ox-LDL, thiobarbituric acid reactive substances, paraoxonase activity or hs-CRP. OxLDL and Lipoprotein(a) were increased in untreated chronic kidney disease. Angiotensin II AT1 receptor expression inversely correlated with renal function (R(2)=0.15, P<0.03) and Lipoprotein(a) but not with the other parameters. Treatment with RAS blockers and statins normalized blood pressure and cholesterol, however it did not correct enhanced leukocyte Angiotensin II AT1 receptor expression. Leukocyte Angiotensin II AT1 receptor expression is inappropriately high in chronic kidney disease, correlates inversely with renal function and does not depend on antihypertensive and lipid-lowering treatment. The uremic environment seems to dominate over previously reported actions of high blood pressure and cholesterol to enhance leukocyte Angiotensin II AT1 receptor expression. PMID- 21640099 TI - Vanillyl nonanoate protects rat gastric mucosa from ethanol-induced injury through a mechanism involving calcitonin gene-related peptide. AB - Previous studies have demonstrated that capsaicin-sensitive sensory nerves are involved in the protection of gastric mucosa against damage by various stimuli and calcitoin gene-related peptide (CGRP) is a potential mediator in this process. This study was performed to explore the effect of vanillyl nonanoate, a capsaicin analog, on ethanol-induced gastric mucosal injury and the possible underlying mechanisms. A rat model of gastric mucosal injury was induced by oral administration of acidified ethanol and gastric tissues were collected for analysis of gastric ulcer index, cellular apoptosis, the activities of caspase-3, catalase and superoxide dismutase (SOD), levels of CGRP, TNF-alpha and malondialdehyde (MDA). The results showed that acute administration of ethanol significantly increased gastric ulcer index concomitantly with increased cellular apoptosis, caspase-3 activity, TNF-alpha and MDA levels as well as decreased activities of catalase and SOD. Pretreatment with 1mg/kg vanillyl nonanoate significantly attenuated ethanol-induced gastric mucosal injury and cellular apoptosis accompanied by increase of CGRP expression, and SOD activity and decrease of caspase-3 activity, TNF-alpha and MDA levels. The effects of vanillyl nonanoate were inhibited by capsazepine, an antagonist of capsaicin receptor. Our results suggested that vanillyl nonanoate was able to protect the gastric mucosa against ethanol-induced gastric mucosal injury. The underlying mechanism is related to stimulation of CGRP release and subsequent suppression of ethanol induced inflammatory reaction, cellular apoptosis and oxidative stress. PMID- 21640100 TI - Effects of aging and hypertension on the participation of endothelium-derived constricting factor (EDCF) in norepinephrine-induced contraction of rat femoral artery. AB - Endothelium-dependent contraction elicited by high concentrations of acetylcholine was described in hypertensive as well as in aged normotensive rats. The contribution of endothelium-derived constricting factor (EDCF) to norepinephrine-induced contraction is still unknown. We aimed to compare EDCF participation to norepinephrine-induced arterial contraction in spontaneously hypertensive rats (SHR) and aged normotensive Wistar-Kyoto (WKY) rats. Femoral arteries from either adult (7-months-old) or aged (14-months-old) animals were placed in myograph and norepinephrine-induced concentration-response curves were recorded under control conditions and in the presence of indomethacin (cyclooxygenase inhibitor, 10(-5) mol/l) or L-NNA (NO synthase inhibitor, 10(-4) mol/l) or both. Norepinephrine-induced concentration-response curve was enhanced in SHR compared to WKY rats, but concentration-response curve of aged WKY rats was similar to those of adult SHR. Cyclooxygenase inhibition largely attenuated concentration-response curves in all groups. However, this effect was greater in aged WKY rats and adult SHR compared to adult WKY rats. NO synthase inhibition augmented norepinephrine-induced contraction in arteries of adult WKY rats, but not in arteries from aged WKY rats or adult SHR. The combined administration of L NNA and indomethacin had no additive effects on concentration-response curves. EDCF contribution to norepinephrine-induced contractions of arteries was considerably greater in adult SHR (80+/-3%) and aged WKY rats (86+/-2%) compared to adult WKY rats (35+/-10%). The inhibition of NO synthase augmented EDCF contribution to norepinephrine-induced contraction only in arteries from adult WKY rats (76+/-9%). We conclude that EDCF contribution to norepinephrine-induced contraction of conduit arteries is similarly enhanced in adult hypertensive and aged normotensive rats. PMID- 21640101 TI - Selection of alkaline phosphatase-positive induced pluripotent stem cells from human amniotic fluid-derived cells by feeder-free system. AB - Generation of induced pluripotent stem (iPS) cells from somatic cells has been successfully achieved by ectopic expression of four transcription factors, Oct4, Sox2, Klf4 and c-Myc, also known as the Yamanaka factors. In practice, initial iPS colonies are picked based on their embryonic stem (ES) cell-like morphology, but often may go on to fail subsequent assays, such as the alkaline phosphate (AP) assay. In this study, we co-expressed through lenti-viral delivery the Yamanaka factors in amniotic fluid-derived (AF) cells. ES-like colonies were picked onto a traditional feeder layer and a high percentage AF-iPS with partial to no AP activity was found. Interestingly, we obtained an overwhelming majority of fully stained AP positive (AP+) AF-iPS colonies when colonies were first seeded on a feeder-free culture system, and then transferred to a feeder layer for expansion. Furthermore, colonies with no AP activity were not detected. This screening step decreased the variation seen between morphology and AP assay. We observed the AF-iPS colonies grown on the feeder layer with 28% AP+ colonies, 45% AP partially positive (AP+/-) colonies and 27% AP negative (AP-) colonies, while colonies screened by the feeder-free system were 84% AP+ colonies, 16% AP+/- colonies and no AP- colonies. The feeder-free screened AP+ AF-iPS colonies were also positive for pluripotent markers, OCT4, SOX2, NANOG, TRA-1-60, TRA-1-81, SSEA-3 and SSEA-4 as well as having differentiation abilities into three germ layers in vitro and in vivo. In this study, we report a simplistic, one-step method for selection of AP+ AF-iPS cells via feeder-free screening. PMID- 21640102 TI - MAML1 regulates cell viability via the NF-kappaB pathway in cervical cancer cell lines. AB - The Notch signaling pathway plays important roles in tumorigenesis in a context dependent manner. In human cervical cancer, alterations in Notch signaling have been reported, and both tumor-suppressing and tumor-promoting roles of Notch signaling have been proposed; however, the precise molecular mechanisms governing these roles in cervical cancer remain controversial. MAML is a transcriptional co activator originally identified by its role in Notch signaling. Recent evidence suggests that it also plays a role in other signaling pathways, such as the p53 and beta-catenin pathways. MAML is required for stable formation of Notch transcriptional complexes at the promoters of Notch target genes. Chromosomal translocations affecting MAML have been shown to promote tumorigenesis. In this study, we used a truncated dominant-negative MAML1 (DN-MAML) to investigate the role of MAML in HPV-positive cervical cancer cell lines. Three human cervical cancer cell lines (HeLa, SiHa and CaSki) expressed all Notch receptors and the Notch target genes Hes1 and MAML1. Among these 3 cell lines, constitutive appearance of cleaved Notch1 was found only in CaSki cells, which suggests that Notch1 is constitutively activated in this cell line. Gamma secretase inhibitor (GSI) treatment, which suppresses Notch receptor activation, completely abrogated this form of Notch1 but had no effect on cell viability. Overexpression of DN MAML by retroviral transduction in CaSki cells resulted in significant decreases in the mRNA levels of Hes1 and Notch1 but had no effects on the levels of MAML1, p53 or HPV E6/E7. DN-MAML expression induced increased viability of CaSki cells without any effect on cell cycle progression or cell proliferation. In addition, clonogenic assay experiments revealed that overexpression of DN-MAML resulted in increased colony formation compared to the overexpression of the control vector. When the status of the NF-kappaB pathway was investigated, CaSki cells overexpressing DN-MAML exhibited loss of phospho-IkappaBalpha, decreased total IkappaBalpha and nuclear localization of NF-kappaB p65, which suggests that the NF-kappaB pathway is hyperactivated. Furthermore, increased level of cleaved Notch1 was detected when DN-MAML was expressed. When DN-MAML-overexpressing cells were treated with GSI, significantly decreased cell viability was observed, indicating that inhibition of Notch signaling using GSI treatment and DN-MAML expression negatively affects cell viability. Taken together, targeting Notch signaling using DN-MAML and GSI treatment may present a novel method to control cell viability in cervical cancer cells. PMID- 21640104 TI - Optical coherence tomography (OCT) measurements in black and white children with large cup-to-disc ratios. AB - Children with large optic nerve head cups often pose diagnostic difficulty due to concern over possible glaucoma. This study's purpose was to evaluate optical coherence tomography (OCT) measurement of the peripapillary retinal nerve fiber layer (RNFL) and macular thickness in healthy eyes of black and white children, comparing values for eyes with large cup-to-disc ratios against those with small cup-to-disc ratios (normal controls). Using Stratus OCT (OCT 3) (Carl Zeiss Meditec, Dublin, CA) a fast macular thickness and fast RNFL 3.4 protocol were performed on normal eyes of normal children. Included were children (aged 5-17 years) with normal ophthalmic examinations, drawn from data generated during an ongoing study of OCT in children. Excluded were eyes with corrected vision below 20/20, spherical equivalent >5 diopters, intraocular pressure >21 mm Hg, abnormal optic nerve rim, prematurity, family history of glaucoma. Clinical examination identified children with large cup-disc-ratios (0.5-0.8), and each was matched for age and race to a "normal control" with a small cup-to-disc ratio (<0.5). OCT 3 analyses were compared between eyes with optic nerve heads showing large cup-to disc ratios (>=0.5) vs. those showing small cup-to-disc ratios (<0.5) for different sections of the macula and the peripapillary RNFL thickness. Analyses included OCT values and optic cup grading for 162 eyes of 162 children. White children with large cup-to-disc ratios had thinner mean RNFL thickness and macular volume compared to those with small cup-to-disc ratios (average RNFL = 100 +/- 9 vs. 110 +/- 8 MUm, respectively, p = 0.0001, macular volume = 6.84 +/- 0.41 vs. 7.03 +/- 0.33 mm(3), respectively, p = 0.0186). On the other hand, black children with large cup-to-disc ratios had an equivalent mean RNFL thickness and macular volume compared to those with small cup-to-disc ratios average RNFL = 107 +/- 10 vs. 107 +/- 8 for both groups, macular volume = 6.81 +/- 0.34 vs. 6.85 +/- 0.28 respectively. Positive equivalence testing assumed a clinically significant difference of 10 MUm for RNFL, and 0.2 mm(3) for macular volume. We report that white children with large cup-to-disc ratios have evidence of reduced/thinner RNFL and macular volume versus white children with smaller cup-to-disc ratios, as assessed by OCT. By contrast, this distinction was not found for black children in the present study. PMID- 21640103 TI - Decreased cell adhesion promotes angiogenesis in a Pyk2-dependent manner. AB - Angiogenesis is regulated by both soluble growth factors and cellular interactions with the extracellular matrix (ECM). While cell adhesion via integrins has been shown to be required for angiogenesis, the effects of quantitative changes in cell adhesion and spreading against the ECM remain less clear. Here, we show that angiogenic sprouting in natural and engineered three dimensional matrices exhibited a biphasic response, with peak sprouting when adhesion to the matrix was limited to intermediate levels. Examining changes in global gene expression to determine a genetic basis for this response, we demonstrate a vascular endothelial growth factor (VEGF)-induced upregulation of genes associated with vascular invasion and remodeling when cell adhesion was limited, whereas cells on highly adhesive surfaces upregulated genes associated with proliferation. To explore a mechanistic basis for this effect, we turned to focal adhesion kinase (FAK), a central player in adhesion signaling previously implicated in angiogenesis, and its homologue, proline-rich tyrosine kinase 2 (Pyk2). While FAK signaling had some impact, our results suggested that Pyk2 can regulate both gene expression and endothelial sprouting through its enhanced activation by VEGF in limited adhesion contexts. We also demonstrate decreased sprouting of tissue explants from Pyk2-null mice as compared to wild type mice as further confirmation of the role of Pyk2 in angiogenic sprouting. These results suggest a surprising finding that limited cell adhesion can enhance endothelial responsiveness to VEGF and demonstrate a novel role for Pyk2 in the adhesive regulation of angiogenesis. PMID- 21640105 TI - Susceptibility to re-infection in C57BL/6 mice with recombinant strains of Toxoplasma gondii. AB - This work reports results of re-infection of BALB/c and C57BL/6 mice with different recombinant strains of Toxoplasma gondii. Mice were prime-infected with the non-virulent D8 strain and challenged with virulent strains. PCR-RFLP of cS10 A6 genetic marker of T. gondii demonstrated that BALB/c mice were re-infected with the EGS strain, while C57BL/6 mice were re-infected with the EGS and CH3 strains. Levels of IFN-gamma and IL-10 after D8 prime-infection were lower in C57BL/6 than in BALB/c mice. Brain inflammation after D8 prime-infection was more intense in C57BL/6 than in BALB/c mice. It was shown that re-infection depends on mice lineage and genotype of the strain used in the challenge. PMID- 21640106 TI - Efficacy of Leishmania donovani ribosomal P1 gene as DNA vaccine in experimental visceral leishmaniasis. AB - The acidic ribosomal proteins of the protozoan parasites have been described as prominent antigens during human disease. We present here data showing the molecular cloning and protective efficacy of P1 gene of Leishmania donovani as DNA vaccine. The PCR amplified complete ORF cloned in either pQE or pVAX vector was used either as peptide or DNA vaccine against experimentally induced visceral leishmaniasis in hamsters. The recombinant protein rLdP1 was given along with Freund's adjuvant and the plasmid DNA vaccine, pVAX-P1 was used alone either as single dose or double dose (prime and boost) in different groups of hamsters which were subsequently challenged with a virulent dose of 1*10(7) L. donovani (MHOM/IN/DD8/1968 strain) promastigotes by intra-cardiac route. While the recombinant protein rLdP1 or DNA vaccine pVAX-P1 in single dose format were not found to be protective, DNA vaccine in a prime-boost mode was able to induce protection with reduced mortality, a significant (75.68%) decrease in splenic parasite burden and increased expression of Th1 type cytokines in immunized hamsters. Histopathology of livers and spleens from these animals showed formation of mature granulomas with compact arrangement of lymphocytes and histiocytes, indicating its protective potential as vaccine candidate. PMID- 21640107 TI - Dynamic regulation of PCNA ubiquitylation/deubiquitylation. AB - Proliferating Cell Nuclear Antigen (PCNA) ubiquitylation plays a crucial role in maintaining genomic stability during DNA replication. DNA damage stalling the DNA replication fork induces PCNA ubiquitylation that activates DNA damage bypass to prevent the collapse of DNA replication forks that could potentially produce double-strand breaks and chromosomal rearrangements. PCNA ubiquitylation dictates the mode of bypass depending on the level of ubiquitylation; monoubiquitylation and polyubiquitylation activate error-prone translesion synthesis and error-free template switching, respectively. Due to the error-prone nature of DNA damage bypass, PCNA ubiquitylation needs to be tightly regulated. Here, we review the molecular mechanisms to remove ubiquitin from PCNA including the emerging role of USP1 and ELG1 in this fascinating process. PMID- 21640108 TI - Destroying the ring: Freeing DNA from Ku with ubiquitin. AB - The Ku heterodimer, consisting of the proteins Ku70 and Ku80, is the central component of the non-homologous end joining (NHEJ) pathway of double strand break (DSB) repair. Ku is able to recognize and bind a DSB by virtue of its ring-like structure. Both pre-repair and topologically trapped post-repair Ku heterodimers are thought to be inhibitory to multiple cellular processes. Thus, a regulated mechanism for the removal of Ku from chromatin was predicted to exist. Recent evidence shows that Ku80 is removed from DNA through a ubiquitin-mediated process. Similar processes have been shown to be involved in the regulated dissociation of a host of other proteins from chromatin, and this appears to be a general and conserved mechanism for the regulation of chromatin-associated factors. A potential mechanism for this pathway is discussed. PMID- 21640109 TI - Unusual case of an upset stomach. Diagnosis: Traumatic diaphragmatic rupture with intrathoracic gastric herniation. PMID- 21640110 TI - Megacolon from Chagas disease in an ancient Texan. Diagnosis: Chagas disease causing mega-disease, in this case megacolon. PMID- 21640111 TI - A man with a testicular mass and a colon stenosis. Diagnosis: Metastatic pancreatic adenocarcinoma to the gonads. PMID- 21640112 TI - Inflammation promotes the loss of adeno-associated virus-mediated transgene expression in mouse liver. AB - BACKGROUND & AIMS: Non-self transgenes delivered to mouse liver via adeno associated virus (AAV) are expressed stably due to the induction of immune tolerance. However, such transgene expression has been reported to be lost in higher-order primates. We investigated whether inflammatory processes, which likely differ between species, impact the stability of transgene expression. METHODS: We developed a mouse model that mimics a scenario in which a subject that has received hepatic AAV-mediated gene transfer develops subsequent, vector unrelated, systemic inflammation. RESULTS: Inflammation eliminated previously stable expression of transgenes delivered by AAV; the limited tissue destruction and persistence of AAV genomes implicated pathways besides the cytotoxic T-cell response. Tumor necrosis factor-a down-regulated expression of the transgene from the AAV, indicating a role for similar inflammatory cytokines in such loss of transgene expression. CONCLUSIONS: Inflammation can block AAV-mediated expression of transgenes in mouse liver. The presence of inflammation might therefore affect hepatic expression of transgenes from viral vectors in humans. PMID- 21640113 TI - An unusual case of obstructive jaundice. Diagnosis: Metastatic melanoma. PMID- 21640114 TI - Echogenic shadow in the common bile duct. Diagnosis: Biliary ascariasis. PMID- 21640115 TI - An unusual case of intraluminally growing esophageal tumor. Diagnosis: Carcinosarcoma of the esophagus. PMID- 21640116 TI - BMPR1A mutations in hereditary nonpolyposis colorectal cancer without mismatch repair deficiency. PMID- 21640117 TI - Bleeding after bagging. Diagnosis: Gastric rupture and massive pneumoperitoneum secondary to barotrauma from bag ventilation. PMID- 21640118 TI - SNAIL regulates interleukin-8 expression, stem cell-like activity, and tumorigenicity of human colorectal carcinoma cells. AB - BACKGROUND & AIMS: Some cancer cells have activities that are similar to those of stem cells from normal tissues, and cell dedifferentiation correlates with poor prognosis. Little is known about the mechanisms that regulate the stem cell-like features of cancer cells; we investigated genes associated with stem cell-like features of colorectal cancer (CRC) cells. METHODS: We isolated colonospheres from primary CRC tissues and cell lines and characterized their gene expression patterns by microarray analysis. We also investigated the biological features of the colonosphere cells. RESULTS: Expanded CRC colonospheres contained cells that expressed high levels of CD44 and CD166, which are markers of colon cancer stem cells, and had many features of cancer stem cells, including chemoresistance and radioresistance, the ability to initiate tumor formation, and activation of epithelial-mesenchymal transition (EMT). SNAIL, an activator of EMT, was expressed at high levels by CRC colonospheres. Overexpression of Snail in CRC cells induced most properties of colonospheres, including cell dedifferentiation. Two hundred twenty-seven SNAIL-activated genes were up-regulated in colonospheres; gene regulatory networks centered around interleukin (IL)-8 and JUN. Blocking IL-8 expression or activity disrupted SNAIL-induced stem cell-like features of colonospheres. We observed that SNAIL activated the expression of IL8 by direct binding to its E3/E4 E-boxes. In CRC tissues, SNAIL and IL-8 were coexpressed with the stem cell marker CD44 but not with CD133 or CD24. CONCLUSIONS: In human CRC tissues, SNAIL regulates expression of IL-8 and other genes to induce cancer stem cell activities. Strategies that disrupt this pathway might be developed to block tumor formation by cancer stem cells. PMID- 21640119 TI - Measuring corticosterone in seabird egg yolk and the presence of high yolk gestagen concentrations. AB - Large inter-species differences have been found in yolk corticosterone amounts in avian eggs. While some studies have failed to detect significant amounts of corticosterone, in other species high amounts have been recorded, such as in a recent study of southern rockhopper penguins Eudyptes chrysocome chrysocome. However, attention has been drawn recently to the fact that many antibodies for corticosterone measurement cross-react with other steroids present in the yolk. In particular, progesterone and related substances can occur in yolk in high concentrations, such that also low cross-reactions of corticosterone assays may lead to measurement errors. We thus performed high-performance liquid chromatographic (HPLC) analyses of yolk extracts and determined the concentration of immunoreactive corticosterone, as well as cross-reacting progesterone and cortisol in egg yolks of southern rockhopper penguins and imperial shags Phalacrocorax atriceps albiventer. We found that high gestagen concentrations in the yolk result in large measurement errors for yolk corticosterone, even when the cross-reactivity seems small. This was observed for both species. We further found species-specific differences in the actual corticosterone amounts present in the egg yolks. PMID- 21640120 TI - Genetic and electron-microscopic characterization of Rickettsiella bacteria from the manuka beetle, Pyronota setosa (Coleoptera: Scarabaeidae). AB - Larvae of manuka beetles, Pyronota spp. (Coleoptera: Scarabaeidae) cause pasture damage in New Zealand by feeding on the roots of grasses. Surveys for potential biocontrol agents revealed a putative disease, expressed as whitened larvae of one of the outbreak species, Pyronota setosa. Microbial diagnosis indicated an intracoelomic, intracellular infection, and intracellular bacteria have been identified with subcellular structures characteristic of infection by Rickettsiella-like microorganisms. These bacteria were rod-shaped, often slightly bent with a mean of 628 nm in length and 220 nm in width. Numerous associated protein crystals of variable size and shape occurred within round to oval shaped "giant bodies" either singly or as clusters of smaller crystals. Molecular phylogenetic analysis based on 16S ribosomal RNA and signal recognition particle receptor (FtsY) encoding sequences demonstrates that the manuka beetle pathogen belongs to the taxonomic genus Rickettsiella. Therefore, the pathotype designation 'Rickettsiella pyronotae' is proposed to refer to this organism. Moreover, genetic analysis makes it likely that--on the basis of the currently accepted organization of the genus Rickettsiella--this new pathotype should be considered a synonym of the nomenclatural type species, Rickettsiella popilliae. PMID- 21640121 TI - Improvement of left ventricular diastolic function induced by beta-blockade: a comparison between nebivolol and metoprolol. AB - OBJECTIVES: Enhanced adrenergic drive is involved in the development of left ventricular (LV) diastolic dysfunction observed in metabolic syndrome (MS). Thus, beta-blockers might improve LV dysfunction observed in MS, but whether this occurs is unknown. METHODS: We assessed in Zucker fa/fa rats the effects of short (5 days) and long-term (90 days) metoprolol ('pure' beta-blockade; 80 mg/kg/day) or nebivolol (beta-blocker with vasodilating properties; 5mg/kg/day) treatment on LV hemodynamics and remodeling, as well as the long-term effects on coronary and peripheral endothelial dysfunction. RESULTS: At identical degree of beta(1) receptor blockade, metoprolol and nebivolol decreased heart rate to the same extent and preserved cardiac output via increased stroke volume. None of the beta blockers, either after long- or short-term administration, modified LV end systolic pressure-volume relation. Both beta-blockers reduced, after long-term administration, LV end-diastolic pressure, Tau and end-diastolic pressure-volume relation, and this was associated with reduced LV collagen density, but not heart weight. Similar hemodynamic effects were also observed after short-term nebivolol, but not short-term metoprolol. These short-term effects of nebivolol were abolished by NO synthase inhibition. At the vascular level, nebivolol, and to a lesser extend metoprolol, improved NO dependent coronary vasorelaxation, which was abolished by NO synthase inhibition. CONCLUSIONS: In a model of MS, the beta-blockers metoprolol and nebivolol improve to the same extent LV hemodynamics, remodeling and diastolic function, but nebivolol prevent more markedly endothelium dependent vasorelaxation involving a more marked enhancement of NO bio-availability. PMID- 21640122 TI - EPR spectra and molecular dynamics agree that the nucleotide pocket of myosin V is closed and that it opens on binding actin. AB - We have used EPR spectroscopy and computational modeling of nucleotide-analog spin probes to investigate conformational changes at the nucleotide site of myosin V. We find that, in the absence of actin, the mobility of a spin-labeled diphosphate analog [spin-labeled ADP (SLADP)] bound at the active site is strongly hindered, suggesting a closed nucleotide pocket. The mobility of the analog increases when the MV.SLADP complex (MV=myosin V) binds to actin, implying an opening of the active site in the A.MV.SLADP complex (A=actin). The probe mobilities are similar to those seen with myosin II, despite the fact that myosin V has dramatically altered kinetics. Molecular dynamics (MD) simulation was used to understand the EPR spectra in terms of the X-ray database. The X-ray structure of MV.ADP.BeFx shows a closed nucleotide site and has been proposed to be the detached state. The MV.ADP structure shows an open nucleotide site and has been proposed to be the A.MV.ADP state at the end of the working powerstroke. MD simulation of SLADP docked in the closed conformation gave a probe mobility comparable to that seen in the EPR spectrum of the MV.SLADP complex. The simulation of the open conformation gave a probe mobility that was 35-40 degrees greater than that observed experimentally for the A.MV.SLADP state. Thus, EPR, X ray diffraction, and computational analysis support the closed conformation as a myosin V state that is detached from actin. The MD results indicate that the MV.ADP crystal structure, which may correspond to the strained actin-bound post powerstroke conformation resulting from head-head interaction in the dimeric processive motor, is superopened. PMID- 21640123 TI - Structural basis for electron and methyl-group transfer in a methyltransferase system operating in the reductive acetyl-CoA pathway. AB - Several anaerobic acetogenic, methanogenic, hydrogenogenic, and sulfate-reducing microorganisms are able to use the reductive acetyl-CoA (Wood-Ljungdahl) pathway to convert CO2 into biomass. The reductive acetyl-CoA pathway consists of two branches connected by the Co/Fe-containing corrinoid iron-sulfur protein (CoFeSP), which transfers a methyl group from a methyltransferase (MeTr)/methyltetrahydrofolate (CH3-H4 folate) complex to the reduced Ni-Ni-[4Fe 4S] cluster (cluster A) of acetyl-CoA synthase. We investigated the CoFeSP and MeTr couple of the hydrogenogenic bacterium Carboxydothermus hydrogenoformans and show that the two proteins are able to catalyze the methyl-group transfer reaction from CH3-H4 folate to the Co(I) center of CoFeSP. We determined the crystal structures of both proteins. The structure of CoFeSP includes the previously unresolved N-terminal domain of the large subunit of CoFeSP, revealing a unique four-helix-bundle-like architecture in which a [4Fe-4S] cluster is shielded by hydrophobic amino acids. It further reveals that the corrinoid and the [4Fe-4S] cluster binding domains are mobile, which is mandatory for the postulated electron transfer between them. Furthermore, we solved the crystal structures of apo-MeTr, CH3-H4-folate-bound MeTr, and H4-folate-bound MeTr, revealing a substrate-induced closure of the CH3-H4 folate binding cavity of MeTr. We observed three different conformations of Asn200 depending on the substrate bound in the active site, demonstrating its conformational modulation by hydrogen-bonding interactions with the substrate. The observed flexibility could be essential to stabilize the transition state during methyl-group transfer. The conformational space and role of Asn200 are likely conserved in homologous cobalamin-dependent MeTrs such as methionine synthase. PMID- 21640124 TI - Consequences of stabilizing the natively disordered f helix for the folding pathway of apomyoglobin. AB - The F helix region of sperm whale apomyoglobin is disordered, undergoing conformational fluctuations between a folded helical conformation and one or more locally unfolded states. To examine the effects of F helix stabilization on the folding pathway of apomyoglobin, we have introduced mutations to augment intrinsic helical structure in the F helix of the kinetic folding intermediate and to increase its propensity to fold early in the pathway, using predictions based on plots of the average area buried upon folding (AABUF) derived from the primary sequence. Two mutant proteins were prepared: a double mutant, P88K/S92K (F2), and a quadruple mutant, P88K/A90L/S92K/A94L (F4). Whereas the AABUF for F2 predicts that the F helix will not fold early in the pathway, the F helix in F4 shows a significantly increased AABUF and is therefore predicted to fold early. Protection of amide protons by formation of hydrogen-bonded helical structure during the early folding events has been analyzed by pH-pulse labeling. Consistent with the AABUF prediction, many of the F helix residues for F4 are significantly protected in the kinetic intermediate but are not protected in the F2 mutant. F4 folds via a kinetically trapped burst-phase intermediate that contains stabilized secondary structure in the A, B, F, G, and H helix regions. Rapid folding of the F helix stabilizes the central core of the misfolded intermediate and inhibits translocation of the H helix back to its native position, thereby decreasing the overall folding rate. PMID- 21640125 TI - Spontaneous similarity discrimination in the evolution of cooperation. AB - The similarity discrimination effect occurs when a single gene or gene cluster causes its carriers to display both a variable phenotypic trait and a behavioural predisposition to cooperate preferentially with recognisably similar carriers. We distinguish this from the greenbeard effect, in which cooperation evolves through fixed phenotypic tags and genetically linked cooperative behaviour with others displaying the same tag. Our agent-based simulations show that the evolution of cooperation through similarity discrimination, in contrast to the greenbeard effect, does not depend on population viscosity or other restrictive conditions. Similarity discrimination evolves spontaneously in well mixed populations, not only in the Prisoner's Dilemma game but also across a range of different binary choice strategic interactions, provided that agents can distinguish reliably between similar and dissimilar co-players. PMID- 21640126 TI - The influence of cytokinin-auxin cross-regulation on cell-fate determination in Arabidopsis thaliana root development. AB - Root growth and development in Arabidopsis thaliana are sustained by a specialised zone termed the meristem, which contains a population of dividing and differentiating cells that are functionally analogous to a stem cell niche in animals. The hormones auxin and cytokinin control meristem size antagonistically. Local accumulation of auxin promotes cell division and the initiation of a lateral root primordium. By contrast, high cytokinin concentrations disrupt the regular pattern of divisions that characterises lateral root development, and promote differentiation. The way in which the hormones interact is controlled by a genetic regulatory network. In this paper, we propose a deterministic mathematical model to describe this network and present model simulations that reproduce the experimentally observed effects of cytokinin on the expression of auxin regulated genes. We show how auxin response genes and auxin efflux transporters may be affected by the presence of cytokinin. We also analyse and compare the responses of the hormones auxin and cytokinin to changes in their supply with the responses obtained by genetic mutations of SHY2, which encodes a protein that plays a key role in balancing cytokinin and auxin regulation of meristem size. We show that although shy2 mutations can qualitatively reproduce the effect of varying auxin and cytokinin supply on their response genes, some elements of the network respond differently to changes in hormonal supply and to genetic mutations, implying a different, general response of the network. We conclude that an analysis based on the ratio between these two hormones may be misleading and that a mathematical model can serve as a useful tool for stimulate further experimental work by predicting the response of the network to changes in hormone levels and to other genetic mutations. PMID- 21640129 TI - Tool-use observation makes far objects ready-to-hand. AB - Previous evidence has shown that active tool-use remaps agents' reaching space with far objects being perceived as reachable and graspable. To date, however, there is no evidence that tool-use observation might also be effective in reaching space remapping. The present six experiments show that not only performing but also observing tool actions may extend the representation of reaching space, useful for grasping objects. In addition, like active tool-use, tool-use observation also impacts on visual distance judgment. Interestingly, these effects only occurred when observers shared the same action potentialities with the agent, i.e., while passively holding a tool compatible with the goal and the spatial range of the observed action. The present findings demonstrate that observing someone else acting with a tool may actually shape the way we map the objects and the space around us, suggesting that such a mapping could provide us with a keystone for coordinating and integrating our actions with those of others. PMID- 21640127 TI - Heterotrimeric G proteins, focal adhesion kinase, and endothelial barrier function. AB - Ligands by binding to G protein coupled receptors (GPCRs) stimulate dissociation of heterotrimeric G proteins into Galpha and Gbetagamma subunits. Released Galpha and Gbetagamma subunits induce discrete signaling cues that differentially regulate focal adhesion kinase (FAK) activity and endothelial barrier function. Activation of G proteins downstream of receptors such as protease activated receptor 1 (PAR1) and histamine receptors rapidly increases endothelial permeability which reverses naturally within the following 1-2 h. However, activation of G proteins coupled to the sphingosine-1-phosphate receptor 1 (S1P1) signal cues that enhance basal barrier endothelial function and restore endothelial barrier function following the increase in endothelial permeability by edemagenic agents. Intriguingly, both PAR1 and S1P1 activation stimulates FAK activity, which associates with alteration in endothelial barrier function by these agonists. In this review, we focus on the role of the G protein subunits downstream of PAR1 and S1P1 in regulating FAK activity and endothelial barrier function. PMID- 21640128 TI - Intravenous nicotine self-administration and cue-induced reinstatement in mice: effects of nicotine dose, rate of drug infusion and prior instrumental training. AB - Intravenous nicotine self-administration is the most direct measure of nicotine reinforcement in laboratory animals, but this procedure has proven difficult to establish in mice. We found that stable responding for nicotine in C57BL6/J mice was facilitated by prior instrumental training for food reward, initial exposure of mice to a lower unit dose of nicotine (0.03 mg kg(-1) per infusion) before access to higher doses, a slower rate of drug delivery (3-s versus 1-s infusion), consistency in schedule of daily testing, and low extraneous noise during testing. Under these conditions, we found that mice lever-pressed for nicotine (0.03-0.4 mg kg(-1) per infusion; 60-min test sessions) under a fixed-ratio 5 time-out 20-s (FR5TO20) reinforcement schedule and consumed the drug according to an inverted 'U'-shaped dose-response curve. Mice switched their responding onto a previously non-reinforced lever to continue earning nicotine infusions when the active/inactive lever assignment was reversed. The nicotinic acetylcholine receptor (nAChR) antagonist mecamylamine decreased responding for nicotine, but not food rewards, verifying that nAChRs regulate nicotine self-administration in mice. The cue-light paired with nicotine delivery did not support responding when delivered independently of nicotine infusions, further verifying that mice responded selectivity for the drug. Nicotine-seeking responses extinguished when nicotine infusions and the cue-light were withheld, and exposure to the cue-light reinstated responding. Finally, mice without prior instrumental food training acquired stable responding for nicotine under the FR5TO20 schedule, but required a greater number of sessions. These data demonstrate that nicotine is an effective reinforcer in mice and establish conditions under which the drug is reliably self-administered by mice. PMID- 21640130 TI - Uptake, distribution and depuration of paralytic shellfish toxins from Alexandrium minutum in Australian greenlip abalone, Haliotis laevigata. AB - Farmed greenlip abalone Haliotis laevigata were fed commercial seaweed-based food pellets or feed pellets supplemented with 8 * 105 Alexandrium minutum dinoflagellate cells g-1 (containing 12 +/- 3.0 MUg STX-equivalent 100 g-1, which was mainly GTX-1,4) every second day for 50 days. Exposure of abalone to PST supplemented feed for 50 days did not affect behaviour or survival but saw accumulation of up to 1.6 MUg STX-equivalent 100 g-1 in the abalone foot tissue (muscle, mouth without oesophagus and epipodial fringe), which is ~50 times lower than the maximum permissible limit (80 MUg 100 g-1 tissue) for PSTs in molluscan shellfish. The PST levels in the foot were reduced to 0.48 MUg STX-equivalent 100 g-1 after scrubbing and removal of the pigment surrounding the epithelium of the epipodial fringe (confirmed by both HPLC and LC-MS/MS). Thus, scrubbing the epipodial fringe, a common procedure during commercial abalone canning, reduced PST levels by ~70%. Only trace levels of PSTs were detected in the viscera (stomach, gut, heart, gonad, gills and mantle) of the abalone. A toxin reduction of approximately 73% was observed in STX-contaminated abalone held in clean water and fed uncontaminated food over 50 days. The low level of PST uptake when abalone were exposed to high numbers of A. minutum cells over a prolonged period may indicate a low risk of PSP poisoning to humans from the consumption of H. laevigata that has been exposed to a bloom of potentially toxic A. minutum in Australia. Further research is required to establish if non-dietary accumulation can result in significant levels of PSTs in abalone. PMID- 21640131 TI - Recombinant conotoxin, TxVIA, produced in yeast has insecticidal activity. AB - Conotoxins are a diverse collection of more than 50,000 peptides produced by predatory marine snails of the genus Conus in order to immobilize their prey. Many conotoxins modulate the activity of ion channels, and show high specificity to their targets; as a result, some have valuable pharmaceutical applications. However, obtaining active peptide is difficult and to date has only been achieved though natural collection, chemical synthesis, or the use of prokaryotic expression systems, which often have the disadvantage of requiring subsequent steps to correctly fold the peptide. This paper reports the production of a conotoxin, TxVIA from Conus textile, as a biologically active recombinant protein, using the yeast Pichia pastoris as expression host. The presence of the pro-peptide was found to be necessary for the expression of biologically active conotoxin. We also show that TxVIA is not, as previously reported, mollusc specific, but also shows insecticidal activity when injected into lepidopteran (cabbage moth) and dipteran (house fly) larvae. In contrast, recombinant TxVIA was not found to be molluscicidal to the grey field slug Deroceras reticulatum. PMID- 21640132 TI - A physico-chemical and biological study of novel chitosan-chloroquinoline derivative for biomedical applications. AB - This paper describes an elegant cross-linking technique for the preparation of chitosan-chloroquinoline derivative by using a greener technique. Chitosan solution in aqueous acetic acid was treated with 2-chloroquinoline-3-carbaldehyde solution to form hydrogel; the resulting hydrogel was subjected to solvent exchange. Combining the results of FTIR and XRD confirmed that 2-chloroquinoline 3-carbaldehyde have been reacted to chitosan. The morphology of the derivative was investigated by X-ray diffraction (XRD) and scanning electron microscopy (SEM). The thermal stability of the derivative was examined by thermogravimetric analysis (TGA). The photoluminescence (PL) spectra of chitosan-chloroquinoline derivative show red-shifted emission maximum. The microbiological screening has demonstrated the antimicrobial activity of the derivative against bacteria viz. Staphylococcus aureus, Escherichia coli and Candida albicans. The obtained results showed that the chitosan-chloroquinoline derivative might be a promising candidate for novel antimicrobial agents for biomedical applications. PMID- 21640133 TI - Enhancement of Astragalus polysaccharide on the immune responses in pigs inoculated with foot-and-mouth disease virus vaccine. AB - The effects of Astragalus polysaccharides (APS) on the immune response in pigs immunized with foot-and-mouth disease virus (FMDV) vaccine were investigated. Fifteen pigs were randomly divided into five groups. Four groups were vaccinated with a FMDV inactivated vaccine. Pigs in three experimental groups were administered varying doses of APS (APS1, 5mg/kg; APS2, 10mg/kg; APS3, 20mg/kg). The influence of APS on the number of CD3(+)CD4(-)CD8(+) cytotoxic T cells, CD3(+)CD4(+)CD8(+) T helper memory cells, and CD3(-)CD4(-)CD8(+) natural killer cells among peripheral blood lymphocytes (PBL) in the three APS groups were significant compared to the vaccine group. In vitro stimulation of PBL by Con A and LPS in APS groups induced a stronger proliferative response at 2 and 6 weeks post-inoculation (PI). APS markedly increased the titer of FMDV-specific antibody in a dose-dependent manner, and up-regulated mRNA expression of IFN-gamma and IL 6. APS could potentially be used as an immunomodulator for a FMDV vaccine and provide better protection against FMDV. PMID- 21640134 TI - Cationic polyelectrolytes-lipases complexes formation as tool for recovery of these enzymes from their natural sources. AB - In the present paper the formation of complexes between positively charge polyelectrolyte (polyethyleneimine and chitosan) and Candida rugosa lipase from a crude extract and porcine lipase from pancreas commercial homogenate preparations were analyzed. The solubility of lipases-cationic polyelectrolytes formation was dependent on: polyelectrolyte densities electrical charge, polyelectrolyte and enzyme concentrations and salts present in the solution. The lipase was recovered from the non-soluble complex by adding of NaCl at a given pH. Although the polyelectrolytes did not affect lipase biological activity, both of them produced good enzyme recovery (>90%); however, purification factors were low. This methodology appears to be a good previous prepurification and concentration method, using, low-cost polymers, allows the design of a purification method where the protein of interest is present in a large volume with respect to the small amount of polyelectrolyte added. PMID- 21640135 TI - Two pitfalls of BOLD fMRI magnitude-based neuroimage analysis: non-negativity and edge effect. AB - BOLD fMRI is accepted as a noninvasive imaging modality for neuroimaging and brain mapping. A BOLD fMRI dataset consists of magnitude and phase components. Currently, only the magnitude is used for neuroimage analysis. In this paper, we show that the fMRI-magnitude-based neuroimage analysis may suffer two pitfalls: one is that the magnitude is non-negative and cannot differentiate positive from negative BOLD activity; the other is an edge effect that may manifest as an edge enhancement or a spatial interior dip artifact at a local uniform BOLD region. We demonstrate these pitfalls via numeric simulations using a BOLD fMRI model and also via a phantom experiment. We also propose a solution by making use of the fMRI phase image, the counterpart of the fMRI magnitude. PMID- 21640136 TI - A low cost fMRI-compatible tracking system using the Nintendo Wii remote. AB - It is sometimes necessary during functional magnetic resonance imaging (fMRI) experiments to capture different movements made by the subjects, e.g. to enable them to control an item or to analyze its kinematics. The aim of this work is to present an inexpensive hand tracking system suitable for use in a high field MRI environment. It works by introducing only one light-emitting diode (LED) in the magnet room, and by receiving its signal with a Nintendo Wii remote (the primary controller for the Nintendo Wii console) placed outside in the control room. Thus, it is possible to take high spatial and temporal resolution registers of a moving point that, in this case, is held by the hand. We tested it using a ball and racket virtual game inside a 3 Tesla MRI scanner to demonstrate the usefulness of the system. The results show the involvement of a number of areas (mainly occipital and frontal, but also parietal and temporal) when subjects are trying to stop an object that is approaching from a first person perspective, matching previous studies performed with related visuomotor tasks. The system presented here is easy to implement, easy to operate and does not produce important head movements or artifacts in the acquired images. Given its low cost and ready availability, the method described here is ideal for use in basic and clinical fMRI research to track one or more moving points that can correspond to limbs, fingers or any other object whose position needs to be known. PMID- 21640137 TI - Dynamic linear model analysis of optical imaging data acquired from the human neocortex. AB - The amount of light absorbed and scattered by neocortical tissue is altered by neuronal activity. Imaging of intrinsic optical signals (ImIOS), a technique for mapping these activity-evoked optical changes with an imaging detector, has the potential to be useful for both clinical and experimental investigations of the human neocortex. However, its usefulness for human studies is currently limited because intraoperatively acquired ImIOS data is noisy. To improve the reliability and usefulness of ImIOS for human studies, it is desirable to find appropriate methods for the removal of noise artifacts and its statistical analysis. Here we develop a Bayesian, dynamic linear modeling approach that appears to address these problems. A dynamic linear model (DLM) was constructed that included cyclic components to model the heartbeat and respiration artifacts, and a local linear component to model the activity-evoked response. The robustness of the model was tested on a set of ImIOS data acquired from the exposed cortices of six human subjects illuminated with either 535nm or 660nm light. The DLM adequately reduced noise artifacts in these data while reliably preserving their activity-evoked optical responses. To demonstrate how these methods might be used for intraoperative neurosurgical mapping, optical data acquired from a single human subject during direct electrical stimulation of the cortex were quantitatively analyzed. This example showed that the DLM can be used to provide quantitative information about human ImIOS data that is not available through qualitative analysis alone. PMID- 21640138 TI - A pilot analytic study of a research-level, lower-cost human papillomavirus 16, 18, and 45 test. AB - The analytic performance of a low-cost, research-stage DNA test for the most carcinogenic human papillomavirus (HPV) genotypes (HPV16, HPV18, and HPV45) in aggregate was evaluated among carcinogenic HPV-positive women, which might be used to decide who needs immediate colposcopy in low-resource settings ("triage test"). We found that HPV16/18/45 test agreed well with two DNA tests, a GP5+/6+ genotyping assay (Kappa = 0.77) and a quantitative PCR assay (at a cutpoint of 5000 viral copies) (Kappa = 0.87). DNA sequencing on a subset of 16 HPV16/18/45 positive and 16 HPV16/18/45 negative verified the analytic specificity of the research test. It is concluded that the HPV16/18/45 assay is a promising triage test with a minimum detection of approximately 5000 viral copies, the clinically relevant threshold. PMID- 21640139 TI - Preparation and evaluation of the 1st international standard for the quantitation of HIV-2 RNA in plasma. AB - An international standard for the quantitation of HIV-2 RNA in plasma samples was developed. A collaborative study involving 29 laboratories from 15 countries was carried out in order to evaluate HIV-2 RNA candidate materials for use with nucleic acid-based tests (NATs). Candidate reference standards consisted of duplicate copies of two HIV-2 genotype A viruses, HIV-2 CAM2 and HIV-2 ROD and were coded S1-S4. Each laboratory assayed all four candidates on at least three separate occasions and data were collated and analysed at NIBSC. Of the data sets returned the majority were from qualitative assays. All assays detected both candidate standards with the exception of one commercial assay, the Nuclisens Easy Q, which was designed primarily for HIV-1 detection which did not detect HIV 2 CAM2 but showed good detection of HIV-2 ROD. This highlighted possible cross reactivity with HIV-2 ROD with some NAT primer/probe combinations; as a result the HIV-2 CAM2 material was established as the 1st international standard for HIV 2 RNA with an assigned unitage of 1000 International Units (IU) per ampoule and is available upon request from the National Institute for Biological Standardisation and Control (NIBSC) (www.nibsc.ac.uk). PMID- 21640140 TI - Novel post-translational modifications of the hemagglutinin and neuraminidase proteins of avian influenza virus expressed by Kluyveromyces lactis. AB - Avian influenza virus (AIV) is an enveloped virus with segmented RNA that belongs to the Orthomyxoviridae. Recently, avian influenza virus isolates have not only posed a significant threat to the poultry industry but also serious public health concerns. The full-length viral hemagglutinin (HA), neuraminidase (NA) or both genes were inserted into the yeast Kluyveromyces lactis genome to allow for secreted expression. Both hemagglutinin and neuraminidase activities were demonstrated for the expressed proteins. Based on PNGase F digestion and immunoassays, N-glycosidically linked high mannose or hybrid-type carbohydrate chains on the HA protein are predominant. It is noteworthy that when co expression of the HA and NA proteins was carried out, the NA protein was able to react with the HA protein, resulting in deglycosylation in a manner similar to PNGase F digestion. Such post-translational modifications in the HA and NA proteins of AIV are described for the first time. PMID- 21640141 TI - Phosphorylated c-Jun and Fra-1 induce matrix metalloproteinase-1 and thereby regulate invasion activity of 143B osteosarcoma cells. AB - Osteosarcoma is the most common primary malignancy of bone and patients often develop pulmonary metastases. Despite the advances in surgical and medical management, the mechanisms underlying human osteosarcoma progression and metastasis remain to be elucidated. Gene expression profiles were compared by the cDNA microarray technique between two different human osteosarcoma sublines, MNNG/HOS and 143B, which differ greatly in spontaneous pulmonary metastatic potential. Here we report an enhanced expression of matrix metalloproteinase (MMP)-1 in the highly metastatic human osteosarcoma cell line 143B. Moreover, the in vitro invasion activity of 143B cells was MMP-1-dependent. The activator protein (AP)-1 binding site in the MMP-1 gene promoter was required for the constitutive expression of MMP-1 in 143B cells. Two AP-1 components, c-Jun and Fra-1, were phosphorylated, and bound to the AP-1 binding site of the MMP-1 promoter in 143B cells. Activated c-Jun and Fra-1 were essential for MMP-1 gene expression in 143B cells. Mitogen-activated protein kinase pathways including the c-Jun NH(2)-terminal kinase and the extracellular signal-regulated kinase activate c-Jun and Fra-1 and thereby regulate c-Jun/Fra-1 mediated events, establishing the mitogen-activated protein kinase/AP-1/MMP-1 axis as important in 143B cells. These data suggest that MMP-1 plays a central role in osteosarcoma invasion. Accordingly, MMP-1 might be a biomarker and therapeutic target for invasive osteosarcomas and pulmonary metastases. PMID- 21640142 TI - Development of a new tool to improve gene transfer frequency calculations. AB - Gene transfer frequency can be determined experimentally on plates, but the methods currently in use do not discriminate between independent transfers and clonal multiplication of initial transformants. In order to overcome this bias, we engineered an Acinetobacter baylyi population in which cells differed by a specific molecular signature and used it as recipient in transformation experiments. Our results suggest that a corrective factor of 0.52 should be applied in order to accurately report natural transformation when using the plate counting method. PMID- 21640143 TI - Reported fatigue, difficulty, and cardiovascular response to an auditory mental arithmetic challenge. AB - Undergraduates scoring low and high on a questionnaire measure of relatively extended fatigue were presented four versions of an auditory mental arithmetic challenge, ranging in difficulty from low to impossibly high. Among Low Fatigue participants, blood pressure and heart rate responses assessed during the work periods first rose and then fell with difficulty. Among High Fatigue participants, blood pressure responses remained low across difficulty conditions, while heart rate responses rose weakly from the low- to the moderate difficulty condition and then declined. Findings are discussed in terms of a recent interactional analysis of fatigue influence on cardiovascular response. PMID- 21640144 TI - Lactate fraction dependent mechanical properties of semitransparent poly(lactate co-3-hydroxybutyrate)s produced by control of lactyl-CoA monomer fluxes in recombinant Escherichia coli. AB - In order to evaluate the mechanical properties of poly(lactate-co-3 hydroxybutyrate) [P(LA-co-3HB)] and its correlation with the LA fraction, P(LA-co 3HB)s with a variety of LA fractions were prepared using recombinant Escherichia coli expressing the LA-polymerizing enzyme and monomer supplying enzymes. The LA overproducing mutant E. coli JW0885 with a pflA gene disruption was used for the LA-enriched polymer production. The LA fraction was also varied by jar-fermentor based fine-regulation of the anaerobic status of the culture conditions, resulting in LA fractions ranging from 4 to 47 mol%. In contrary to the opaque P(3HB) film, the copolymer films attained semitransparency depending on the LA fraction. Young's modulus values of the P(LA-co-3HB)s (from 148 to 905 MPa) were lower than those of poly(lactic acid) (PLA) (1020 MPa) and P(3HB) (1079 MPa). In addition, the value of elongation at break of the copolymer with 29 mol% LA reached 150%. In conclusion, P(LA-co-3HB)s were found to be a comparatively pliable and flexible material, differing from both of the rigid homopolymers. PMID- 21640145 TI - The vesicle size of DDA:TDB liposomal adjuvants plays a role in the cell-mediated immune response but has no significant effect on antibody production. AB - The use of cationic liposomes as experimental adjuvants for subunit peptide of protein vaccines is well documented. Recently the cationic liposome CAF01, composed of dimethyldioctadecylammonium (DDA) and trehalose dibehenate (TDB), has entered Phase I clinical trials for use in a tuberculosis (TB) vaccine. CAF01 liposomes are a heterogeneous population with a mean vesicle size of 500 nm; a strong retention of antigen at the injection site and a Th1-biassed immune response are noted. The purpose of this study was to investigate whether CAF01 liposomes of significantly different vesicle sizes exhibited altered pharmacokinetics in vivo and cellular uptake with activation in vitro. Furthermore, the immune response against the TB antigen Ag85B-ESAT-6 was followed when various sized CAF01 liposomes were used as vaccine adjuvants. The results showed no differences in vaccine (liposome or antigen) draining from the injection site, however, significant differences in the movement of liposomes to the popliteal lymph node were noted. Liposome uptake by THP-1 vitamin D3 stimulated macrophage-like cells did not show a liposome size-dependent pattern of uptake. Finally, whilst there were no significant differences in the IgG1/2 regardless of the liposome size used as a delivery vehicle for Ag85B-ESAT-6, vesicle size has a size dependent effect on cell proliferation and IL-10 production with larger liposomes (in excess of 2 MUm) promoting the highest proliferation and lowest IL-10 responses, yet vesicles of ~500 nm promoting higher IFN-gamma cytokine production from splenocytes and higher IL-1beta at the site of injection. PMID- 21640146 TI - APN/CD13-targeting as a strategy to alter the tumor accumulation of liposomes. AB - Targeting angiogenic vasculature has been validated as a viable approach for cancer imaging and therapy. The tumor vasculature-specific ligand asparagine glycine-arginine (NGR) peptide targets the isoform of aminopeptidase N (APN, also referred to as CD13) that is expressed on the endothelial cells in angiogenic vessels such as the neovasculature in tumors. APN/CD13 has become widely recognized as a rational target for therapeutic development and several NGR conjugated agents are now in pre-clinical and clinical development. In the current study, a CT image-based approach is used to evaluate the in vivo performance of several NGR conjugated liposome formulations that vary in terms of NGR density and PEG spacer arm length. Indeed, for the first time it is demonstrated that CT imaging can be used for quantitative and longitudinal assessment of the pharmacokinetics and biodistribution of an actively targeted liposome formulation containing an iodinated agent. In comparison to conventional methods, the CT image guided drug delivery approach enables visualization of the intratumoral distribution of liposomes and quantification of the fraction of tumor occupied by the vesicles over time. This study is the first to use CT for molecular imaging. PMID- 21640147 TI - A multiplexed electrospray process for single-step synthesis of stabilized polymer particles for drug delivery. AB - While conventional methods for biodegradable particle production rely predominately on batch, emulsion preparation methods, an alternative process based on multiplexed electrospray (ES) can offer distinct advantages. These include enhanced encapsulation efficiency of hydrophilic and hydrophobic agents, scale-up potential, tight control over particle size and excellent particulate reproducibility. Here we developed a well-controlled ES process to synthesize coated biodegradable polymer particles. We demonstrate this process with the Poly(DL-lactic-co-glycolic acid) system encapsulating amphiphilic agents such as doxorubicin (DOX), Rhodamine B (RHO(B)) and Rhodamine B octadecyl ester perchlorate (RHO(BOEP)). We show that in a single-step flow process particles can be made encapsulating the agent with high efficiency and coated either with emulsifiers that stabilize them in solution or that may facilitate further functionalization for targeted drug delivery. The coating process allows for the surface modification of the particles without further changes in particle size or morphology, and with minimal loss of drug (>94% encapsulation efficiency). This synthesis technique is well suited for massive scale-up using microfabricated, multiplexed arrays consisting of multiple electrospray nozzles operating in parallel. A simple analytical model of the diffusion of the encapsulated agent within the polymer reveals two distinct phases in the cumulative release profile: a first phase in which the release is dominated by diffusion and a second phase with a slower release related to the erosion of the polymer matrix. The first, diffusion-driven stage is highly affected by particle agglomeration properties, whereas the second one shows a much less pronounced dependence on particle size. Modeling suggests that the size of the particles will substantially influence the initial burst in both the percentage of drug released and the rate at which it is released. It will also affect to a smaller extent the secondary slow and sustained release. Our study highlights the importance of tight control over particle size and morphology and the avoidance of particle aggregation for control over the release kinetics and formulation repeatability. PMID- 21640149 TI - Optimization of a novel and improved thermosensitive liposome formulated with DPPC and a Brij surfactant using a robust in vitro system. AB - The combination of thermosensitive liposomes and local heating has been shown to improve anticancer drug delivery in both animal models and human patients. The lyso-lipid temperature sensitive liposomes (LTSL) consisting of DPPC, MSPC and DSPE-PEG(2000) is currently under evaluation in clinical trials. We hypothesized that Brij surfactants resembling the chemical structures of MSPC and DSPE PEG(2000) could be utilized for generating a thermosensitive formulation with DPPC. Here, we report using a robust in vitro system to efficiently screen a series of liposomal candidates composed of DPPC and a Brij surfactant for thermosensitive delivery of doxorubicin. The data indicated that the optimal acyl chain length of the surfactant was between C(16) and C(18) with a saturated carbon chain, a PEG repeating unit ranging between 10 and 100 and a molecule weight above 600Da. The linking chemistry between the acyl chain and the PEG chain did not influence thermosensitivity. In the panel of surfactants tested, Brij78 was optimal and could be incorporated into the liposomes by the thin film hydration or the post-insertion method with an optimal range of 1 to 8mol%. Doxorubicin was incorporated into the formulation by pH gradient with >95% loading efficiency at drug/lipid of 1/20 (w/w). The transition temperature of the Brij78-liposomes was slightly lower than that of LTSL (41 v.s. 41.5 degrees C), leading to enhanced drug release at the low end of the hyperthermic temperatures (40 degrees C) with similar stability at 37 degrees C, which was confirmed by cell based assays. Finally, the Brij78-liposomes and LTSL displayed comparable blood compatibility with mild hemolytic activity. This in vitro system allowed for efficient screening and optimization to produce an optimal formulation. PMID- 21640148 TI - Kinetics of skin resealing after insertion of microneedles in human subjects. AB - Over the past decade, microneedles have been shown to dramatically increase skin permeability to a broad range of compounds by creating reversible microchannels in the skin. However, in order to achieve sustained transdermal drug delivery, the extent and duration of skin's increased permeability needs to be determined. In this study, we used electrical impedance spectroscopy to perform the first experiments in human subjects to analyze the resealing of skin's barrier properties after insertion of microneedles. Microneedles having a range of geometries were studied in conjunction with the effect of occlusion to test the hypothesis that increasing microneedle length, number, and cross-sectional area together with occlusion leads to an increase in skin resealing time that can exceed one day. Results indicated that in the absence of occlusion, all microneedle treated sites recovered barrier properties within 2 h, while occluded sites resealed more slowly, with resealing windows ranging from 3 to 40 h depending on microneedle geometry. Upon subsequent removal of occlusion, the skin barrier resealed rapidly. Longer microneedles, increased number of needles, and larger cross-sectional area demonstrated slower resealing kinetics indicating that microneedle geometry played a significant role in the barrier resealing process. Overall, this study showed that pre-treatment of skin with microneedles before applying an occlusive transdermal patch can increase skin permeability for more than one day, but nonetheless allow skin to reseal rapidly after patch removal. PMID- 21640150 TI - Presence of metabolic syndrome and cardiovascular risk factors in adolescents and University students in Crete (Greece), according to different levels of snack consumption. AB - The aim of this study was to investigate the presence of cardiovascular and metabolic syndrome (MetSyn) risk factors in adolescents and University students in Crete (Greece), according to different levels of snack consumption. 237 adolescents (14.2-16.3 years) and 196 University students (19.5-31.5 years) participated during 2002/3 and 2004/5. Dietary intake (24-h dietary recalls), cardiorespiratory fitness, smoking habits, time spent watching television, anthropometric, blood pressure and biochemical measurements were assessed in all subjects. Adolescents consumed snacks more often than University students (62% vs. 49.5%, P < 0.05) and had significantly higher total energy intake (8629 vs. 7641 kJ, P < 0.05) and energy intake from snacks (2537 vs. 1767 kJ, P < 0.001). Snack consumers had higher energy intake than non-consumers in both groups. No relationship was found between snack consumption and MetSyn risk factors, cardiorespiratory fitness and smoking. However, University students who consumed snacks had 15.36 times higher risk of having >= 2 MetSyn risk factors than adolescent snack consumers, had considerably higher LDL-cholesterol levels, total: HDL-cholesterol ratio and watched more television than non-consumers. These findings emphasize the need for developing effective nutrition interventions to promote healthier snack choices in adolescents and young adults with characteristics similar to our sample, in order to prevent cardiovascular risk later in adulthood. PMID- 21640151 TI - Purification and biochemical characterization of antioxidant peptide from horse mackerel (Magalaspis cordyla) viscera protein. AB - In the present study, a peptide having high antioxidant properties was isolated from horse mackerel viscera protein, Magalaspis cordyla. In vitro gastrointestinal digestion was employed to obtain potential protein hydrolysate and was subjected to consecutive chromatographic methods using fast protein liquid chromatography (FPLC) connected to diethyl amino ethyl (DEAE) anion exchange column and Sephadex G-25 gel filtration column. The activity of the fractions was tested against DPPH and hydroxyl radicals and the isolated peptide showed 89.2 and 59.1 percentage of scavenging. The amino acid sequence of purified peptide was determined using ESI-MS/MS as Ala-Cys-Phe-Leu (518.5 Da), it exhibited high activity against polyunsaturated fatty acid (PUFA) peroxidation than that of natural antioxidant, alpha-tocopherol. PMID- 21640152 TI - Trivalent influenza vaccine and febrile adverse events in Australia, 2010: clinical features and potential mechanisms. AB - INTRODUCTION: Increased numbers of children presenting with febrile adverse events following trivalent influenza vaccine (TIV) were noted in Australia in 2010. We describe the epidemiology and clinical features of the adverse events and explore the biological basis for the adverse events using an in vitro model. MATERIALS AND METHODS: Children presenting to a tertiary paediatric hospital in 2010 with adverse events within 72 h of TIV were retrospectively reviewed. Demographics, clinical features, physiological variables and outcomes were examined. Plasma cytokine and chemokine levels were examined in a subgroup of children with vaccine-related febrile convulsions. Peripheral blood mononuclear cells of age-matched children were stimulated with different TIV preparations. Inflammatory cytokine and chemokine analysis was performed on cultured supernatants. RESULTS: Vaccine-related febrile adverse events were identified in 190 children. Most occurred in healthy children (median age: 1.5 years) within 12 h of vaccination. Twenty-eight (14.7%) required hospital admission. High temperature >=39.0 degrees C (101/190; 53%), vomiting (120/190; 63%) and convulsions (38/190; 20%) were common. All children presenting had received Fluvax((r)) or Fluvax Junior((r)). In the in vitro model, IFN-alpha, IL-1beta, IL 6, IL-10, IP-10 and MIP-1alpha levels were significantly higher when measured at 6 and 24 h in cultures stimulated with Fluvax((r)) compared with alternative 2010 TIV preparations. CONCLUSIONS: Numerous febrile adverse events (including febrile seizures) were observed following Fluvax((r)) or Fluvax Junior((r)) in 2010. Clear differences in cytokine production were observed when peripheral blood mononuclear cells were stimulated with Fluvax((r)) compared with alternate TIV preparations. Increased awareness of these potential adverse events is required to ensure earlier detection and prevention in the future. PMID- 21640153 TI - Dimensional correlates of poor insight in obsessive-compulsive disorder. AB - BACKGROUND: Cross-sectional studies have associated poor insight in patients with obsessive-compulsive disorder (OCD) with increased OCD symptom severity, earlier age of onset, comorbid depression, and treatment response. The goal of this current study was to examine the relationship between dimensions of OCD symptomatology and insight in a large clinical cohort of Brazilian patients with OCD. We hypothesized that poor insight would be associated with total symptom severity as well as with hoarding symptoms severity, specifically. METHODS: 824 outpatients underwent a detailed clinical assessment for OCD, including the Yale Brown Obsessive-Compulsive Scale (Y-BOCS), the Dimensional Yale-Brown Obsessive Compulsive Scale (DY-BOCS), the Brown Assessment of Beliefs Scale (BABS), a socio demographic questionnaire, and the Structured Clinical Interview for axis I DSM IV disorders (SCID-P). Tobit regression models were used to examine the association between level of insight and clinical variables of interest. RESULTS: Increased severity of current and worst-ever hoarding symptoms and higher rate of unemployment were associated with poor insight in OCD after controlling for current OCD severity, age and gender. Poor insight was also correlated with increased severity of current OCD symptoms. CONCLUSION: Hoarding and overall OCD severity were significantly but weakly associated with level of insight in OCD patients. Further studies should examine insight as a moderator and mediator of treatment response in OCD in both behavioral therapy and pharmacological trials. Behavioral techniques aimed at enhancing insight may be potentially beneficial in OCD, especially among patients with hoarding. PMID- 21640154 TI - Protective mechanisms of anthocyanins from purple sweet potato against tert-butyl hydroperoxide-induced hepatotoxicity. AB - Anthocyanins have been shown to exert anti-proliferative, anti-inflammatory effects and anti-carcinogenic activity. In the present work, we investigated the protective effects of anthocyanin fraction (AF) from purple sweet potato on tert butyl hydroperoxide (t-BHP)-induced hepatotoxicity in HepG2 cell line and in rat liver. The result showed that the oral pretreatment of AF before t-BHP treatment significantly lowered the serum levels of the hepatic enzyme markers (ALT and AST) and reduced oxidative stress of the liver by evaluation of malondialdehyde and glutathione. Histopathological evaluation of the livers also revealed that AF reduced the incidence of liver lesions. The in vitro result showed that AF significantly reduced t-BHP-induced oxidative injury, as determined by cell cytotoxicity, intracellular glutathione content, lipid peroxidation, reactive oxygen species (ROS) levels, and caspases activation. Also, AF up-regulated antioxidant enzymes including heme oxygenase-1 (HO-1), NAD(P)H:quinone reductase, and glutathione S-transferase. Moreover, AF induced Nrf2 nuclear translocation and Akt and ERK1/2 activation, pathways that are involved in inducing Nrf2 nuclear translocation. Taken together, these results suggest that the protective effects of AF against t-BHP-induced hepatotoxicity may, at least in part, be due to its ability to scavenge ROS and to regulate the antioxidant enzyme HO-1 via the Akt and ERK1/2/Nrf2 signaling pathways. PMID- 21640155 TI - An OGG1 polymorphism is associated with mitochondrial DNA content in pesticide exposed fruit growers. AB - Exposure to pesticides has the capacity to cause mitochondrial dysfunction. An increase mitochondrial DNA (mtDNA) content has also been suggested to relate with DNA damaging agent. In mitochondria, the manganese superoxide dismutase (MnSOD) is a scavenger of reactive oxygen species, and the 8-oxoguanine DNA glycosylase (OGG1) is the major DNA glycosylase for the repair of 8-oxoG lesions. However, the alteration of mtDNA content elicited by pesticide exposure in people with genetic variations in MnSOD or OGG1 has not been investigated. In this study, the mitochondrial to nuclear DNA ratio was quantified in the peripheral blood of 120 fruit growers who experienced pesticide exposure and 106 unexposed controls by real-time quantitative polymerase chain reaction (real-time qPCR). Questionnaires were administered to obtain demographic data and occupational history. The MnSOD and OGG1 genotypes were identified by the PCR based restriction fragment length polymorphism analysis. After adjusting for confounding effects, multiple regression model revealed that subjects experiencing high or low pesticide exposure had a greater mtDNA content than that of controls. The OGG1 Ser-Ser genotype was also associated with an increased mtDNA content. No association between MnSOD genotype and mtDNA content was revealed. Thus, subjects experiencing pesticide exposure had greater mtDNA content and the OGG1 genotype may modulate mtDNA content in pesticide-exposed fruit growers. PMID- 21640156 TI - Invariant local conformation in p22phox p.Y72H polymorphisms suggested by mass spectral analysis of crosslinked human neutrophil flavocytochrome b. AB - The NADPH oxidase of phagocytic leukocytes generates superoxide that plays a critical role in innate immunity and inflammatory responses. The integral membrane protein flavocytochrome b (Cyt b, a.k.a. cytochrome b(558/559)) is the catalytic core of the complex and serves as a prototype for homologs important in regulating signaling networks in a wide variety of animal and plant cells. Our analysis identifies a naturally-occurring Tyr72/His72 polymorphism (p.Y72H) in the p22(phox) subunit of Cyt b at the protein level that has been recognized at the nucleotide level (c.214T > C, formerly C242T) and implicated in cardiovascular disease. In the present study, Cyt b was isolated from human neutrophils and reacted with chemical crosslinkers for subsequent structure analysis by MALDI mass spectrometry. Following mild chemical modification of Cyt b with two pairs of isotopically-differentiated lysine crosslinkers: BS(2)G d(0)/d(4) and BS(3)-d(0)/d(4), the reaction mixtures were digested with trypsin and purified on C(18)ZipTips to generate samples for mass analysis. MALDI analysis of tryptic digests from each of the above reactions revealed a series of masses that could be assigned to p22(phox) residues 68-85, assuming an intra molecular crosslink between Lys71 and Lys78. In addition to the 30 ppm mass accuracy obtained with internal mass calibration, increased confidence in the assignment of the crosslinks was provided by the presence of the diagnostic mass patterns resulting from the isotopically-differentiated crosslinking reagent pairs and the Tyr72/His72 p22(phox) polymorphisms in the crosslinked peptides. This work identifies a novel, low-resolution distance constraint in p22(phox) and suggests that the medically-relevant p.Y72H polymorphism has an invariant structural motif in this region. Because position 72 in p22(phox) lies outside regions identified as interactive with other oxidase components, the structural invariance also provides additional support for maturational differences as the source of the wide variation in observed reactive oxygen species production by cells expressing p.Y72H. PMID- 21640157 TI - Bcl-2 is a better therapeutic target than c-Myc, but attacking both could be a more effective treatment strategy for B-cell lymphoma with concurrent Bcl-2 and c Myc overexpression. AB - OBJECTIVE: The prognosis for diffuse large B-cell lymphomas with concomitant overexpression of c-Myc and Bcl-2 remains dismal; there is an urgent need to clarify the significance of these two oncogenes as therapeutic targets for a more effective treatment strategy. MATERIALS AND METHODS: We established two novel cell lines, KPUM-MS3 and KPUM-UH1, from two chemoresistant patients with diffuse large B-cell lymphomas with concomitant overexpression of c-Myc and Bcl-2, and investigated the significance of c-Myc and Bcl-2 as therapeutic targets. RESULTS: KPUM-MS3 possesses t(14;18)(q32;q21) chromosomal translocation and KPUM-UH1 bcl-2 gene amplification, both of which account for Bcl-2 overexpression. Chromosomal translocation t(8;14)(q24;q34) was found to coexist only in KPUM-UH1, overexpression of pvt-1 messenger RNA was detected only in KPUM-MS3, and reduced expression of miR-143 and miR-145 was identified in both. Working together, these abnormalities can contribute to c-Myc overexpression. Using ABT-263, an inhibitor for Bcl-2, and 10058-F4, an inhibitor for c-Myc, we found that both cell lines were more highly sensitive to cell death as a result of Bcl-2 inhibition than of c-Myc inhibition. When combined with genotoxic agents, ABT-263 exerted additive and/or synergistic cell-killing effects, while 10058-F4 showed, at most, a modest combinatory effect. Importantly, the combination of ABT-263 and 10058-F4 had a synergistic cell-killing effect on both cell lines. CONCLUSIONS: Our data suggest that Bcl-2 is a better therapeutic target than c-Myc, but attacking both Bcl-2 and c-Myc would be an even more effective treatment strategy for diffuse large B cell lymphomas with concurrent Bcl-2 and c-Myc overexpression. PMID- 21640158 TI - Effects of chronic administration of tryptophan with or without concomitant fluoxetine in depression-related and anxiety-like behaviors on adult rat. AB - Depression and anxiety play an important role in decreasing quality of life worldwide. Since tryptophan is a serotonin precursor and low levels of serotonin seems to be related to depression, the effect of oral tryptophan has been investigated for possible potentiation of the action of antidepressant drugs. We investigated the effects of chronically administered tryptophan (50mg/kg/day, p.o.) with or without concomitant fluoxetine (10mg/kg/day, s.c.) on adult rats regarding depression-related and anxiety-like behaviors. Tryptophan levels in cerebrospinal fluid (CSF) were measured 4h after a single administration of daily dosages of chronic treatments. We found that tryptophan increased depressive related behavior, but did not alter anxiety-like behavior. However, fluoxetine decreased depression-related behavior and was anxiogenic. Tryptophan with concomitant fluoxetine did not alter anxiety-like behavior. Moreover, our data suggests that the antidepressant effect of fluoxetine was not enhanced by concomitant administration of tryptophan, which could be associated with increased levels of tryptophan in CSF. Further investigations are needed to elucidate the related mechanisms. PMID- 21640159 TI - Acetylsalicylic acid as an augmentation agent in fluoxetine treatment resistant depressive rats. AB - Clinical studies have reported that adjunctive acetylsalicylic acid (aspirin) therapy is beneficial for patients with treatment resistant depression (TRD). However, there still exist negative epidemiological data on the link between aspirin and depression. Therefore, this study aimed to further investigate whether aspirin can be used as an augmentation agent in fluoxetine treatment resistant depressive rats induced by chronic unpredictable mild stress (CUMS). In this study, the effects of CUMS regimen and antidepressant treatment were assessed by behavioral testing, hippocampal expression of cyclooxygenase-2 (COX 2) and prostaglandin E(2) (PGE(2)). 4-week fluoxetine treatment reversed the behavioral changes in approximately 70-80% depressive rats. That is, 20-30% depressive rats were resistant to fluoxetine. In the hippocampus of fluoxetine treatment resistant depressive rats, a significant upregulation of COX-2 level and PGE(2) concentration was observed. However, in these rats adjunctive aspirin treatment significantly improved the depressive behaviors and downregulated the COX-2 level and PGE(2) concentration in the hippocampus. Thus, our results suggest that aspirin can be served as an effective adjunctive agent in the treatment resistant depression mediated by inhibition of the COX-2 level and PGE(2) concentration. PMID- 21640160 TI - 6-Hydroxydopamine lesions of the ventral tegmental area suppress ghrelin's ability to elicit food-reinforced behavior. AB - While past research suggests that ghrelin stimulates appetite through an action on hypothalamic signaling, recent evidence indicates that the peptide acts via mesotelencephalic dopamine neurons to alter appetitive motivation. In the present study, rats were trained to operantly respond for food on a progressive ratio PR5 schedule until stable breakpoints were established. Ghrelin (30-300 pmol) was then injected directly into the ventral tegmental area (VTA) and the 300 pmol dose was observed to increase breakpoint. The dopaminergic neurotoxin 6 hydroxydopamine (6-OHDA, 6 MUg) was subsequently administered into the VTA resulting in a significant depletion of striatal dopamine. Stable breakpoints were then re-established. When ghrelin's effects were reassessed, the peptide's ability to alter operant responding for food was reliably reduced. Our findings demonstrate that ghrelin induces food-reinforced behavior in the mesotelencephalic reward pathway and that this effect is dependent on intact dopaminergic signaling. We conclude that the metabolic peptide ghrelin interacts with dopamine, within reward circuitry, to modulate appetitive behavior. PMID- 21640161 TI - On the functional diversity of glyceraldehyde-3-phosphate dehydrogenase: biochemical mechanisms and regulatory control. AB - BACKGROUND: New studies provide evidence that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is not simply a classical glycolytic protein of little interest. Instead, it is a multifunctional protein with significant activity in a number of fundamental cell pathways. GAPDH is a highly conserved gene and protein, with a single mRNA transcribed from a unique gene. Control mechanisms must exist which regulate its functional diversity. SCOPE OF REVIEW: This review focuses on new, timely studies defining not only its diverse activities but also those which define the regulatory mechanisms through which those functions may be controlled. The reader is referred to the author's prior review for the consideration of past reports which first indicated GAPDH multiple activities (Sirover, Biochim. Biophys. Acta 1432 (1999) 159-184.) CONCLUSIONS: These investigations demonstrate fundamental roles of GAPDH in vivo, dynamic changes in its subcellular localization, and the importance of posttranslational modifications as well as protein:protein interactions as regulatory control mechanisms. GENERAL SIGNIFICANCE: GAPDH is the prototype "moonlighting" protein which exhibits activities distinct from their classically identified functions. Their participation in diverse cell pathways is essential. Regulatory mechanisms exist which control those diverse activities as well as changes in their subcellular localization as a consequence of those new functions. PMID- 21640162 TI - Brief treatment with heparin-binding EGF-like growth factor, but not with EGF, is sufficient to accelerate epithelial wound healing. AB - BACKGROUND: Heparin-binding EGF-like growth factor (HB-EGF) contains, in contrast to EGF, a domain that binds to negatively charged glycans on cell surfaces and in extracellular matrix. We speculated that a short exposure to HB-EGF induces prolonged biological effects such as healing of wounds after immobilization in tissues. METHODS: Epithelial cell sheets in tissue and corneas in organ culture were treated briefly with HB-EGF or EGF and binding of the growth factors, time course of activation of the EGF receptor, and healing of wounds were compared. RESULTS: Treating human corneal epithelial cells for 2 min with HB-EGF resulted in 8h of detectable activation of the EGF receptor, but activation was much shorter after EGF treatment. A brief treatment with HB-EGF, but not with EGF, induced significant acceleration of healing in wounds in epithelial sheets in tissue and organ culture. Bound HB-EGF was detectable up to 16 h after brief treatments. Neutralizing antibodies added after HB-EGF treatment blocked acceleration of healing, demonstrating the role of bound HB-EGF in accelerating healing. CONCLUSIONS: A brief exposure to HB-EGF, but not to EGF, is sufficient to induce prolonged activation of the EGF receptor and to enhance healing. GENERAL SIGNIFICANCE: Bound HB-EGF can serve as a pool that induces prolonged activation of the EGF receptor. EGF has been used experimentally to treat poorly healing wounds, but the frequent applications that are necessary have hampered its use clinically. The findings imply that HB-EGF may be a useful long-acting alternative to EGF. PMID- 21640164 TI - Sensory gating, inhibition control and child intelligence: an event-related potentials study. AB - The current study explored the relationship among sensory gating, inhibition control and human intelligence in two groups of children with different intellectual levels. A Go-Nogo task was adopted to investigate children's behavioral performances in inhibition control processing, and a paired-click paradigm with event-related potentials (ERP) recording was used to explore children's neural activation during sensory gating processing. The behavioral results showed that the intellectually gifted children committed significantly less commission error rate, which indicated that gifted children had better inhibition control than their average peers. The electrophysiological results showed that the gifted group had lower S2P50/S1P50 amplitude ratio than the average group and illustrated that gifted children had stronger sensory gating. The results of correlation analysis between inhibition control performances and sensory gating showed that children with stronger P50 suppression (lower S2/S1 latency ratio) in the fronto-central area and stronger N100 suppression (lower S2/S1 amplitude ratio) in the frontal and fronto-central areas had shorter reaction time in the Go-Nogo task. Moreover, the correlation patterns between sensory gating and inhibition control were different between two groups of children. The present findings further demonstrated the close relationship among sensory gating, inhibition control and human intelligence in children. PMID- 21640163 TI - S-nitrosylation in the regulation of gene transcription. AB - BACKGROUND: Post-translational modification of proteins by S-nitrosylation serves as a major mode of signaling in mammalian cells and a growing body of evidence has shown that transcription factors and their activating pathways are primary targets. S-nitrosylation directly modifies a number of transcription factors, including NF-kappaB, HIF-1, and AP-1. In addition, S-nitrosylation can indirectly regulate gene transcription by modulating other cell signaling pathways, in particular JNK kinase and ras. SCOPE OF REVIEW: The evolution of S-nitrosylation as a signaling mechanism in the regulation of gene transcription, physiological advantages of protein S-nitrosylation in the control of gene transcription, and discussion of the many transcriptional proteins modulated by S-nitrosylation is summarized. MAJOR CONCLUSIONS: S-nitrosylation plays a crucial role in the control of mammalian gene transcription with numerous transcription factors regulated by this modification. Many of these proteins serve as immunomodulators, and inducible nitric oxide synthase (iNOS) is regarded as a principal mediatiator of NO-dependent S-nitrosylation. However, additional targets within the nucleus (e.g. histone deacetylases) and alternative mechanisms of S-nitrosylation (e.g. GAPDH-mediated trans-nitrosylation) are thought to play a role in NOS-dependent transcriptional regulation. GENERAL SIGNIFICANCE: Derangement of SNO-regulated gene transcription is an important factor in a variety of pathological conditions including neoplasia and sepsis. A better understanding of protein S-nitrosylation as it relates to gene transcription and the physiological mechanisms behind this process is likely to lead to novel therapies for these disorders. This article is part of a Special Issue entitled Regulation of Cellular Processes by S nitrosylation. PMID- 21640165 TI - Impact of methamphetamine on dopamine neurons in primates is dependent on age: implications for development of Parkinson's disease. AB - Methamphetamine is a CNS stimulant with limited therapeutic indications, but is widely abused. Short-term exposure to higher doses, or long-term exposure to lower doses, of methamphetamine induces lasting damage to nigrostriatal dopamine neurons in man and animals. Strong evidence indicates that the mechanism for this detrimental effect on dopamine neurons involves oxidative stress exerted by reactive oxygen species. This study investigates the relative susceptibility of dopamine neurons in mid-gestation, young, and adult (not aged) monkeys to four treatments with methamphetamine over 2 days. Primate dopamine neurons undergo natural cell death at mid-gestation, and we hypothesized that during this event they are particularly vulnerable to oxidative stress. The results indicated that at mid-gestation and in adults, dopamine neurons were susceptible to methamphetamine-induced damage, as indicated by loss of striatal tyrosine hydroxylase (TH) immunoreactivity and dopamine concentration. However, dopamine neurons in young animals appeared totally resistant to the treatment, despite this group having higher brain levels of methamphetamine 3 h after administration than the adults. As a possible explanation for the protection, striatal glial derived neurotrophic factor (GDNF) levels were elevated in young animals 1 week after treatment, but not in adults following methamphetamine treatment. Implications of these primate studies are: (1) the susceptibility of dopamine neurons at mid-gestation to methamphetamine warns against the risk of exposing pregnant women to the drug or oxidative stressors, and supports the hypothesis of Parkinson's disease being associated with oxidative stress during development, (2) elucidation of the mechanism of resistance of dopamine neurons in the young animals to methamphetamine-induced oxidative stress may provide targets for slowing or preventing age- or disease-related loss of adult nigrostriatal dopamine (DA) neurons, and (3) the increased striatal production of GDNF in young animals, but not in adults, in response to methamphetamine, suggests the possibility of an age-related change in the neurotrophic capacity of the striatal dopamine system. PMID- 21640166 TI - A maternal diet supplemented with creatine from mid-pregnancy protects the newborn spiny mouse brain from birth hypoxia. AB - The creatine-phosphocreatine shuttle is essential for the maintenance of cellular ATP, particularly under hypoxic conditions when respiration may become anaerobic. Using a model of intrapartum hypoxia in the precocial spiny mouse (Acomys cahirinus), the present study assessed the potential for maternal creatine supplementation during pregnancy to protect the developing brain from the effects of birth hypoxia. On day 38 of gestation (term is 39 days), the pregnant uterus was isolated and placed in a saline bath for 7.5 min, inducing global hypoxia. The pups were then removed, resuscitated, and cross-fostered to a nursing dam. Control offspring were delivered by caesarean section and recovered immediately after release from the uterus. At 24 h after birth hypoxia, the brains of offspring from dams fed a normal diet showed significant increases in lipid peroxidation as measured by the amount of malondialdehyde. In the cortical subplate, thalamus and piriform cortex there were significant increases in cellular expression of the pro-apoptotic protein BAX, cytoplasmic cytochrome c and caspase-3. When pregnant dams were fed the creatine supplemented diet, the increase in malondialdehyde, BAX, cytochrome c and caspase 3 were almost completely prevented, such that they were not different from control (caesarean delivered) neonates. This study provides evidence that the neuroprotective capacity of creatine in the hypoxic perinatal brain involves abrogation of lipid peroxidation and apoptosis, possibly through the maintenance of mitochondrial function. Further investigation into these mechanisms of protection, and the long term development and behavioural outcomes of such neonates is warranted. PMID- 21640167 TI - Grip force regulates hand impedance to optimize object stability in high impact loads. AB - Anticipatory grip force adjustments are a prime example of the predictive nature of motor control. An object held in precision grip is stabilized by fine adjustments of the grip force against changes in tangential load force arising from inertia during acceleration and deceleration. When an object is subject to sudden impact loads, prediction becomes critical as the time available for sensory feedback is very short. Here, we investigated the control of grip force when participants performed a targeted tapping task with a hand-held object. During the initial transport phase of the movement, load force varied smoothly with acceleration. In contrast, in the collision, load forces sharply increased to very large values. In the transport phase, grip force and load force were coupled in phase, as expected. However, in the collision, grip force did not parallel load force. Rather, it exhibited a stereotyped profile with maximum ~65 ms after peak load at contact. By using catch trials and a virtual environment, we demonstrate that this peak of grip force is pre-programmed. This observation is validated across experimental manipulations involving different target stiffness and directions of movement. We suggest that the central nervous system optimizes stability in object manipulation-as in catching-by regulating mechanical parameters including stiffness and damping through grip force. This study provides novel insights about how the brain coordinates grip force in manipulation involving an object interacting with the environment. PMID- 21640168 TI - Altered brain neurotrophins at birth: consequence of imbalance in maternal folic acid and vitamin B12 metabolism. AB - Folic acid fortification to pregnant women is suggested to mask vitamin B12 deficiency leading to adverse neurologic consequences. The present study examines the effect of maternal folic acid supplementation at normal and excess levels both in the presence and absence of vitamin B12 on levels and expression of brain neurotrophins in Wistar Albino rats. Pregnant female rats were assigned to six dietary groups with varying levels of folic acid and vitamin B12, that is, (NFB: 2 mg folic acid+B12; NFBD: 2 mg folic acid-B12; EFB: 8 mg folic acid+B12; EFBD: 8 mg folic acid-B12; NFBDO: 2 mg folic acid-B12+DHA and EFBDO: 8 mg folic acid B12+DHA). On day 20 of gestation pup brain samples were collected to assess protein and mRNA levels of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF). Brain BDNF protein and mRNA levels were reduced (P<0.01 for both) in the EFBD group as compared to control. However, NGF protein levels were reduced (P<0.05) only in the EFBD group in comparison to EFB and control. Maternal supplementation of DHA improved pup brain NGF protein levels only in the NFBDO (P<0.05) and EFBDO (P<0.05) groups compared to NFBD and EFBD respectively. Our results suggest that maternal micronutrients during pregnancy play an important role in regulating protein and mRNA levels of neurotrophins. Maternal DHA supplementation to a micronutrient imbalanced diet could ameliorate the negative effects only for NGF but not for BDNF. PMID- 21640169 TI - Previous chronic exposure eliminates the conditioning effect of nicotine in rats. AB - Smoking continues to be a major health problem and unfortunately smoking cessation interventions have limited success; the conditioning effects of nicotine and individual differences in tobacco addiction are important factors that underlie this setback. The aim of the current study was to investigate nicotine-induced conditioned place preference (CPP) in male and female rats which were previously exposed to a free choice of oral nicotine or water and showed different preferences for nicotine; subsequently nicotine intake also varied between subjects. Exposure patterns were varied in three experiments to allow for assessing the effect of adult v.s. adolescent exposure. The design of CPP testing enabled testing for the possible confounding effects of withdrawal or tolerance. A total of 150 male and female rats were used in three experiments. The oral nicotine choice was provided for at least six weeks in all experiments. Our results replicate our previous findings that nicotine induces CPP in male, but not female rats not pre-exposed to nicotine. Previous nicotine exposure, irrespective of the amount of nicotine consumed, eliminated the conditioning effects of nicotine in a new context. The diminished CPP response was more pronounced in rats exposed to nicotine as adolescents than those exposed as adults. This reduced responsiveness cannot be explained by tolerance. The neuroplastic changes caused by chronic nicotine administration or the strong conditioning to receiving nicotine in the home cage before CPP testing may underlie the weakened responsiveness. These findings support the well known clinical notion that smoking cessation attempts are more successful in a novel environment, not previously connected with smoking. PMID- 21640170 TI - Genome-wide identification of gibberellins metabolic enzyme genes and expression profiling analysis during seed germination in maize. AB - Gibberellin (GA) is an essential phytohormone that controls many aspects of plant development. To enhance our understanding of GA metabolism in maize, we intensively screened and identified 27 candidate genes encoding the seven GA metabolic enzymes including ent-copalyl diphosphate synthase (CPS), ent-kaurene synthase (KS), ent-kaurene oxidase (KO), ent-kaurenoic acid oxidase (KAO), GA 20 oxidase (GA20ox), GA 3-oxidase (GA3ox), and GA 2-oxidase (GA2ox), using all available public maize databases. The results indicate that maize genome contains three CPS, four KS, two KO and one KAO genes, and most of them are arranged separately on the maize genome, which differs from that in rice. In addition, the enzymes catalyzing the later steps (ZmGA20ox, ZmGA3ox and ZmGA2ox) are also encoded by gene families in maize, but GA3ox enzyme is likely to be encoded by single gene. Expression profiling analysis exhibited that transcripts of 15 GA metabolic genes could be detected during maize seed germination, which provides further evidence for the notion that increased synthesis of active GA in the embryo is required for triggering germination events. Moreover, a variety of temporal genes expression patterns of GA metabolic genes were detected, which revealed the complexity of underlying mechanism for GA regulated seed germination. PMID- 21640171 TI - A poplar DRE-binding protein gene, PeDREB2L, is involved in regulation of defense response against abiotic stress. AB - Dehydration responsive element (DRE)-binding proteins comprise a family of proteins that have important roles in abiotic stress processes. The present study aimed at elucidating the molecular basis of the stress tolerance mechanism in poplar. We isolated a gene, PeDREB2L, from the desert tree Populus euphratica Oliva. PeDREB2L contains a conserved AP2/ERF domain, a nuclear localization signal at the N-terminus and a serine-rich region at the C-terminus. PeDREB2L binds specifically to DRE elements and is expressed ubiquitously in all tissues, including roots, stems, and leaves. Expression of PeDREB2L in leaves increased under dehydration, salt, and abscisic acid stress treatments. Furthermore, PeDREB2L-GFP ectopically expressed in Arabidopsis gave strong green fluorescent protein signals in the nucleus of transgenic Arabidopsis under unstressed conditions and resulted in an improved tolerance against drought and freezing under stress conditions, while the Arabidopsis protein gene DREB2A-GFP used as a control only gave a weak fluorescent. We expected that the PeDREB2L might be useful in improving abiotic stress tolerance in transgenic plants. PMID- 21640172 TI - Molecular evolution of the EGF-CFC protein family. AB - The epidermal growth factor-Cripto-1/FRL-1/Cryptic (EGF-CFC) proteins, characterized by the highly conserved EGF and CFC domains, are extracellular membrane associated growth factor-like glycoproteins. These proteins are essential components of the Nodal signaling pathway during early vertebrate embryogenesis. Homologs of the EGF-CFC family have also been implicated in tumorigenesis in humans. Yet, little is known about the mode of molecular evolution in this family. Here we investigate the origin, extent of conservation and evolutionary relationships of EGF-CFC proteins across the metazoa. The results suggest that the first appearance of the EGF-CFC gene occurred in the ancestor of the deuterostomes. Phylogenetic analysis supports the classification of the family into distinct subfamilies that appear to have evolved through lineage-specific duplication and divergence. Site-specific analyses of evolutionary rate shifts between the two major mammalian paralogous subfamilies, Cripto and Cryptic, reveal critical amino acid sites that may account for the observed functional divergence. Furthermore, estimates of functional divergence suggest that rapid change of evolutionary rates at sites located mainly in the CFC domain may contribute towards distinct functional properties of the two paralogs. PMID- 21640173 TI - Maximum likelihood analysis of mammalian p53 indicates the presence of positively selected sites and higher tumorigenic mutations in purifying sites. AB - The tumor suppressor gene TP53 (p53) maintains genome stability. Mutation or loss of p53 is found in most cancers. Analysis of evolutionary constrains and p53 mutations reveal important sites for concomitant functional studies. In this study, phylogenetic analyses of the coding sequences of p53 from 26 mammals were carried out by applying a maximum likelihood method. The results display two branches under adaptive evolution in mammals. Moreover, each codon of p53 was analyzed by the PAML method for presence of positively selected sites. PAML identified several statistically significant amino acids that undergo positive selection. The data indicates that amino acids responsible for the core functions of p53 are highly conserved, while positively selected sites are predominantly located in the N- and C-terminus of p53. Further analysis of evolutionary pressure and mutations showed the occurrence of more frequent tumorigenic mutations in purifying sites of p53. PMID- 21640174 TI - Identification and characterization of genes encoding two novel LEA proteins in Antarctic and temperate strains of Chlorella vulgaris. AB - An Antarctic strain (NJ-7) of Chlorella vulgaris possesses the same 18S rRNA sequence as that of a temperate strain (UTEX259), but shows significantly higher freezing tolerance than the latter. Suppression subtractive hybridization (SSH) was performed to identify genes of intensified expression in NJ-7 relative to UTEX259. Among the genes identified, Ccor1 and Ccor2, co-organized in the same gene cluster Ccor1-Ccor2-Ccor1-Ccor2, showed much higher expression levels in NJ 7 than in UTEX259 at both 20 degrees C and 4 degrees C. As detected by Northern blot and Western blot analyses, the two genes were cold-inducible in NJ-7 but almost not expressed in UTEX259. Their encoded products are predicted to share 55.7% identity to each other and possess physicochemical characteristics similar to that of late embryogenesis abundant (LEA) proteins in plants. The purified recombinant Ccor1 and Ccor2 showed high heat-stability and could act as cryoprotectants to lactate dehydrogenase in vitro. Based on their expression patterns and protein characteristics, we propose that Ccor1 and Ccor2 are two novel LEA proteins and are related to the greatly enhanced freezing tolerance in the Antarctic strain. PMID- 21640175 TI - Does sex matter? The influence of gender on gastrointestinal physiology and drug delivery. AB - We all respond differently to drugs. Personalised medicine aims to improve efficacy and reduce side effects, and efforts are being made to understand the physiological differences that underlie responses to drugs. Genetics, diet and disease state can be key; however, gender also plays an important role in pharmacokinetics, pharmacodynamics and drug toxicity. Differences in metabolism and clearance of drugs as a consequence of distinct hepatic and renal processes in males and females are now much better understood but little is known about gender differences in the gastrointestinal tract. As the recipient of all orally administered medications, differences at this level can have a major impact on drug delivery and bioavailability; yet these continue to be ignored and insufficiently studied in the context of drug disposition. The aim of this review is to highlight the known gender differences in gut physiology. Clinical case studies are presented, where possible, to illustrate the influence of these differences on drug disposition and gaps in current knowledge are highlighted to encourage further research in this area. PMID- 21640176 TI - Training sound localization in normal hearing listeners with and without a unilateral ear plug. AB - Surprisingly little is known about the ability of adult human listeners to learn to localize sounds in the free field. In this study, we presented broadband noise bursts at 24 equally spaced locations in a 360 degrees horizontal plane in both normal-hearing conditions and when listeners were fitted with a unilateral earplug. Localization improvement was found over the initial four training sessions, prior to plug insertion which produced an immediate and profound impairment in localization, particularly on the side of the plug. Subsequent training with the plug in place over the next 5 days showed continually improving performance (learning) up to the 4th day. Following plug removal, localization immediately returned to pre-plug levels. These results showed that task-specific training can improve localization ability in normal-hearing conditions and that training also improves performance during a unilateral conductive hearing loss. It has been suggested that the process of learning is due to a gradual reweighting of the available cues to develop a new location map. The return to preplug learning performance suggests that the original location map is preserved despite the formation of a new map, and is in agreement with other reported findings. PMID- 21640177 TI - Short-term synaptic plasticity in the auditory brain stem by using in-vivo-like stimulation parameters. AB - Reduced systems such as brain slices offer a powerful approach to study the physiology of auditory neurons in great detail. However, when studying auditory nuclei in reduced systems such as brain slices, especially highly active auditory brain stem nuclei, one has to be aware that the unphysiological lack of activity in the reduced system compared to the in-vivo situation has a number of important effects on the neurons under investigation, and thus on the data that are measured. Most importantly, the lack of chronic activity in the slice preparation has important effects on the properties of short-term plasticity of the synapses. The main purpose of this article is to discuss how spontaneous activity in auditory neurons, or the lack thereof, can affect the data measured. PMID- 21640178 TI - New developments in aminoglycoside therapy and ototoxicity. AB - After almost seven decades in clinical use, aminoglycoside antibiotics still remain indispensible drugs for acute infections and specific indications such as tuberculosis or the containment of pseudomonas bacteria in patients with cystic fibrosis. The review will describe the pathology and pathophysiology of aminoglycoside-induced auditory and vestibular toxicity in humans and experimental animals and explore contemporary views of the mechanisms of cell death. It will also outline the current state of protective therapy and recent advances in the development of aminoglycoside derivatives with low toxicity profiles for antimicrobial treatment and for stop-codon suppression in the attenuation of genetic disorders. PMID- 21640180 TI - Efficacy evaluations of Mimosa pudica tannin isolate (MPT) for its anti-ophidian properties. AB - AIM OF THE STUDY: Evaluations of the anti-snake venom efficacy of Mimosa pudica tannin isolate (MPT) obtained from root of the plant. MATERIALS AND METHOD: MPT was investigated in vitro and in vivo for its efficacy against the venom of Naja kaouthia snake. RESULTS: In vitro: (1) mice injected i.p. with MPT pre-incubated with Naja kaouthia venom at concentrations as low as 0.625 mg/ml showed 100% survival after a 24-h observation period. (2) In the proteomics study, mice injected with MPT pre-incubated with the Naja kaouthia venom showed down regulation of five serum proteins. (3) In the protein-dye-binding study, the percentage of Bradford dye-protein binding showed a reduction relative to the decrease in MPT concentration used to incubate with the venom. In vivo: the results from the animal studies showed that MPT had no in vivo protection against the Naja kaouthia venom (0.875 mg/kg) in four different rescue modes and in an oral pre-treatment experiment. CONCLUSION: The study indicated the promising ability of MPT to neutralize the Naja kaouthia venom in in vitro experiments but fell short in its in vivo potential. As such, the use of Mimosa pudica (Mimosaceae) as therapeutics for snake bites is questionable as all the possible in vivo rescue studies and pre-treatment of the active constituents showed no protection against the affected mice. PMID- 21640179 TI - Anti-epileptic drugs delay age-related loss of spiral ganglion neurons via T-type calcium channel. AB - Loss of spiral ganglion neurons is a major cause of age-related hearing loss (presbycusis). Despite being the third most prevalent condition afflicting elderly persons, there are no known medications to prevent presbycusis. Because calcium signaling has long been implicated in age-related neuronal death, we investigated T-type calcium channels. This family is comprised of three members (Ca(v)3.1, Ca(v)3.2, and Ca(v)3.3), based on their respective main pore-forming alpha subunits: alpha1G, alpha1H, and alpha1I. In the present study, we report a significant delay of age-related loss of cochlear function and preservation of spiral ganglion neurons in alpha1H null and heterozygous mice, clearly demonstrating an important role for Ca(v)3.2 in age-related neuronal loss. Furthermore, we show that anticonvulsant drugs from a family of T-type calcium channel blockers can significantly preserve spiral ganglion neurons during aging. To our knowledge, this is the first report of drugs capable of diminishing age related loss of spiral ganglion neurons. PMID- 21640181 TI - Antidepressant-like effects of the fractions of Xiaoyaosan on rat model of chronic unpredictable mild stress. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Xiaoyaosan (XYS), composed of Radix Bupleuri, Radix Angelicae Sinensis, Radix Paeoniae Alba, Rhizoma Atractylodis Macrocephalae, Poria, Herba Menthae, Rhizoma Zingiberis Recens and Radix Glycyrrhizae, is a valuable traditional Chinese medicine (TCM) which is used for the treatment of depression in China. In the formula, Radix Bupleuri usually serves as the principal drug, Radix Angelicae Sinensis and Radix Paeoniae Alba serve as the ministerial drugs, Rhizoma Atractylodis Macrocephalae, Poria, Herba Menthae and Rhizoma Zingiberis Recens serve as adjunctive drugs, Radix Glycyrrhizae serves as messenger drug, they coordinate with each other and enhance the effect of the formula. In our previous experiments, the antidepressant effect of XYS was revealed. However, the antidepressant part (or component) of this prescription was still obscure. We divided the XYS into five different polar fractions, and explored the antidepressant activity of five different polar fractions to identify the active fraction. MATERIALS AND METHODS: Behavior research and metabonomics method based on (1)H NMR were used for efficacy study of different fractions in chronic unpredictable mild stress (CUMS) model of depression. Rats were divided into 8 groups and drugs were administered during the 21 days model building period. The urine samples of rats were collected overnight (12h) on 21 day and the metabolic profiling of the urine was measured using NMR. Multivariate analysis was also utilized to evaluate the active fraction of XYS. RESULTS: In the behavior research, there were significant difference between the lipophilic fraction group (XY-A) and the model group. In addition, with pattern recognition analysis of urinary metabolites, the results showed a clear separation of the model group and control group, while XY-A group was much closer to the control group in the OSC-PLS score plot. Seven endogenous metabolites contributing to the separation of the model group and control group were detected, while XY-A group regulated the 5 perturbed metabolites showing a tendency of recovering to control group. CONCLUSIONS: The present work suggested that petroleum ether fraction was the most effective fraction, implying that lipophilic components contribute to the antidepressant effect of XYS. PMID- 21640182 TI - Voluntary exercise as an anti-neuroinflammatory therapeutic. PMID- 21640184 TI - Development by environment interactions controlling tryptophan hydroxylase expression. AB - Tryptophan hydroxylase is the rate-limiting enzyme in the biosynthesis of serotonin (5-hydroxytryptamine; 5-HT). Two isoforms of tryptophan hydroxylase, derived from different genes, tph1 and tph2, have been identified. The tph1 isoform is expressed in peripheral tissues, whereas tph2 is brain and neuron specific. Recent studies suggest that tph2 expression and brain serotonin turnover are upregulated in depressed suicide patients, and drug-free depressed patients, respectively. Increased tph2 expression could result from genetic influences, early life developmental influences, adverse experience during adulthood, or interactions among these factors. Studies in rodents support the hypothesis that interactions between early life developmental influences and adverse experience during adulthood play an important role in determining tph2 expression. In this review, we highlight the evidence for the effects of adverse early life experience and stressful experience during adulthood on both tph1 and tph2 expression. PMID- 21640185 TI - Functional organization of the dorsal raphe efferent system with special consideration of nitrergic cell groups. AB - The serotonin (5HT) system of the brain is involved in many CNS functions including sensory perception, stress responses and psychological disorders such as anxiety and depression. Of the nine 5HT nuclei located in the mammalian brain, the dorsal raphe nucleus (DRN) has the most extensive forebrain connectivity and is implicated in the manifestation of stress-related psychological disturbances. Initial investigations of DRN efferent connections failed to acknowledge the rostrocaudal and mediolateral organization of the nucleus or its neurochemical heterogeneity. More recent studies have focused on the non-5HT contingent of DRN cells and have revealed an intrinsic intranuclear organization of the DRN which has specific implications for sensory signal processing and stress responses. Of particular interest are spatially segregated subsets of nitric oxide producing neurons that are activated by stressors and that have unique efferent projection fields. In this regard, both the midline and lateral wing subregions of the DRN have emerged as prominent loci for future investigation of nitric oxide function and modulation of sensory- and stressor-related signals in the DRN and coinciding terminal fields. PMID- 21640183 TI - The serotonergic anatomy of the developing human medulla oblongata: implications for pediatric disorders of homeostasis. AB - The caudal serotonergic (5-HT) system is a critical component of a medullary "homeostatic network" that regulates protective responses to metabolic stressors such as hypoxia, hypercapnia, and hyperthermia. We define anatomically the caudal 5-HT system in the human medulla as 5-HT neuronal cell bodies located in the raphe (raphe obscurus, raphe magnus, and raphe pallidus), extra-raphe (gigantocellularis, paragigantocellularis lateralis, intermediate reticular zone, lateral reticular nucleus, and nucleus subtrigeminalis), and ventral surface (arcuate nucleus). These 5-HT neurons are adjacent to all of the respiratory- and autonomic-related nuclei in the medulla where they are positioned to modulate directly the responses of these effector nuclei. In the following review, we highlight the topography and development of the caudal 5-HT system in the human fetus and infant, and its inter-relationships with nicotinic, GABAergic, and cytokine receptors. We also summarize pediatric disorders in early life which we term "developmental serotonopathies" of the caudal (as well as rostral) 5-HT domain and which are associated with homeostatic imbalances. The delineation of the development and organization of the human caudal 5-HT system provides the critical foundation for the neuropathologic elucidation of its disorders directly in the human brain. PMID- 21640186 TI - Study on in situ and in vivo absorption kinetics of phenytoin by modulating P glycoprotein with verapamil in rats. AB - P-gp is playing significant role in the development of the drug resistance by altering the absorption of drugs. The objective of present investigation was to study the effect of verapamil on the pharmacokinetics of phenytoin in order to evaluate the role of P-glycoprotein (P-gp) in phenytoin absorption. An in situ single pass intestinal perfusion study was carried out to determine the effect of verapamil on the functional status of intestinal P-gp. Phenytoin (30MUM) and propranolol (100MUM) co-perfused with and without verapamil (200MUM) in rat's ileum. An in vivo study, verapamil (25mg/kg, per oral) was administered 2h before phenytoin (30mg/kg, per oral) dosing in male Wistar rats. Plasma samples were collected at 0, 1, 2, 3, 4, 5, 6, 8, 12 and 24h time points from control and treated animals to determine phenytoin concentrations. An in situ single pass intestinal perfusion study indicated phenytoin to be a P-gp substrate and the function of intestinal P-gp was significantly inhibited in presence of verapamil. After per oral phenytoin dosing, the mean area under the plasma concentration time curve (AUC) was found to be 35.03+/-1.83hMUg/ml which was increased significantly, i.e. 62.33+/-2.49hMUg/ml (P<0.001) when verapamil was co administered. Similarly the mean maximum plasma concentration of phenytoin increased from 3.2+/-0.148MUg/ml without verapamil to 6.983+/-0.272MUg/ml (P<0.001) with verapamil. These results are quite stimulating for further development of a clinically useful oral formulation of phenytoin based on P-gp inhibition. PMID- 21640187 TI - Influence of polymer hydrolysis on adjuvant effect of Gantrez(r)AN nanoparticles: implications for oral vaccination. AB - The adjuvant effect of methylvinylether-co-maleic anhydride (Gantrez(r)AN) nanoparticles was investigated during oral vaccination of mice with F4 adhesins of F4-positive Escherichia coli. To differentiate whether the adjuvant effect originated from a nanoparticle effect or a polymer effect, 20 MUg F4 was administered as slightly crosslinked F4-containing nanoparticles (g(F4)(0.01)) or as F4 mixed with slightly crosslinked pure nanoparticles (F4+g(0.01)). The F4 specific immune response was reduced using F4-containing nanoparticles due to complete shielding of F4, whereas oral administration of F4+g(0.01) increased the level of F4-specific antibody-secreting cells (ASC) in the spleen. When repeating the vaccination study after 6months using freshly prepared nanoparticles, the adjuvant effect of F4+g(0.01) was lost due to an altered polymer reactivity caused by partial hydrolysis of anhydride groups of Gantrez(r)AN. Combining F4 with nanoparticles stabilised with a higher crosslinker amount during nanoparticle synthesis (F4+g(0.22)) could overcome the effect of partial polymer hydrolysis, as higher levels of ASC were detected. Hence, an in-depth characterisation of the Gantrez(r)AN polymer is required as stability issues can alter its biological effect during oral vaccination. PMID- 21640188 TI - Glutamate receptor dynamics and protein interaction: lessons from the NMDA receptor. AB - The plasticity of excitatory glutamate synapses emerged over the last decades as a core cellular mechanism for the encoding and processing of various cognitive functions. This property relies in part on the ability to dynamically adjust the content of glutamate receptors in the postsynaptic membrane. Among these receptors, NMDA receptors (NMDAR), which are composed of two obligatory GluN1 and two regulatory GluN2/3 subunits, play a key role in the induction of many forms of plasticity processes. Understanding how NMDAR subtypes are trafficked and regulated in the synapse has thus captured considerable attention. It has emerged that NMDAR synaptic content relies on an equilibrium between intracellular trafficking and rapid lateral diffusion of the receptor within the synaptic area. Here, we review our current understanding of NMDAR trafficking, mostly the ones at the surface membrane, with a specific focus on the role of interacting PDZ containing proteins during the journey of NMDAR to and around the synaptic area. The cellular and molecular lessons obtained from examining NMDAR dynamics and regulation by interacting proteins appear to apply to other ionotropic neurotransmitter receptors, and thus shed new light on the modulation of excitatory, inhibitory, and modulatory transmission. This article is part of a Special Issue entitled 'Neuronal Function'. PMID- 21640189 TI - Expression and purification of myristoylated matrix protein of Mason-Pfizer monkey virus for NMR and MS measurements. AB - Matrix proteins play multiple roles both in early and late stages of the viral replication cycle. Their N-terminal myristoylation is important for interaction with the host cell membrane during virus budding. We used Escherichia coli, carrying N-myristoyltransferase gene, for the expression of the myristoylated His tagged matrix protein of Mason-Pfizer monkey virus. An efficient, single-step purification procedure eliminating all contaminating proteins including, importantly, the non-myristoylated matrix protein was designed. The comparison of NMR spectra of matrix protein with its myristoylated form revealed substantial structural changes induced by this fatty acid modification. PMID- 21640190 TI - Experimental evaluation of support vector machine-based and correlation-based approaches to automatic particle selection. AB - The goal of this study is to evaluate the performance of software for automated particle-boxing, and in particular the performance of a new tool (TextonSVM) that recognizes the characteristic texture of particles of interest. As part of a high throughput protocol, we use human editing that is based solely on class-average images to create final data sets that are enriched in what the investigator considers to be true-positive particles. The Fourier shell correlation (FSC) function is then used to characterize the homogeneity of different single particle data sets that are derived from the same micrographs by two or more alternative methods. We find that the homogeneity is generally quite similar for class-edited data sets obtained by the texture-based method and by SIGNATURE, a cross-correlation-based method. The precision-recall characteristics of the texture-based method are, on the other hand, significantly better than those of the cross-correlation based method; that is to say, the texture-based approach produces a smaller fraction of false positives in the initial set of candidate particles. The computational efficiency of the two approaches is generally within a factor of two of one another. In situations when it is helpful to use a larger number of templates (exemplars), however, TextonSVM scales in a much more efficient way than do boxing programs that are based on localized cross correlation. PMID- 21640191 TI - Activation of the dentate nucleus in a verb generation task: A 7T MRI study. AB - There is increasing evidence of a topographic organization within the human cerebellar cortex for motor and non-motor functions. Likewise, a subdivision of the dentate nucleus in a more dorsal and rostral motor domain and a more ventral and caudal non-motor domain has been proposed by Dum and Strick (2003) based on anatomical studies in monkey. In humans, however, very little is known about topographic organization within the dentate nucleus. Activation of the dentate nucleus in a verb generation task was examined in young and healthy subjects using ultra-highfield 7T functional magnetic resonance imaging (fMRI) with its increase in signal-to-noise ratio. Data of 17 subjects were included in statistical analysis. Subjects were asked to (i) read words (nouns) aloud presented on a screen, (ii) silently read the same nouns, (iii) silently generate the appropriate verbs to the same nouns and (iv) to silently repeat the names of the months. A block design was used. For image processing, a recently developed region of interest (ROI) driven normalization method of the dentate nuclei was applied. Activation related to motor speech (contrast aloud reading minus silent reading) was strongest in the rostral parts of the dentate nucleus. Dorsorostral activations were present bilaterally. Activation related to verb generation (contrast verb generation minus silent reading) was found in the ventrocaudal parts of the dentate nucleus on the right. The present findings are in good accordance with the anatomical data in monkeys and suggest that the human dentate nucleus can be subdivided into a rostral and more dorsal motor domain and a ventrocaudal non-motor domain. PMID- 21640192 TI - Functional neuroimaging of the oculomotor brainstem network in humans. AB - The cortical systems involved in eye movement control in humans have been investigated extensively using fMRI. In contrast, there is virtually no data concerning the functional status of the human oculomotor brainstem nuclei. This lack of evidence has usually been explained by technical constraints of EPI based imaging and anatomical characteristics of the brainstem. Against this assumption, we successfully localised nuclei of the oculomotor system using high-resolution fMRI based on standard EPI sequences in a group of healthy subjects executing reflexive horizontal saccades. A random-effects group analysis revealed task related BOLD increases in the superior colliculus, the oculomotor nucleus, the abducens nucleus and in the paramedian pontine reticular formation. This group analysis was complemented by individual positive findings in up to 94% of single subject analyses. A visual control paradigm led to increased signal levels in the superior colliculus consistent with its visual properties but no corresponding signal changes in other brainstem nuclei. These results are consistent with findings in animal studies and demonstrate the feasibility to detect BOLD signal increases associated with oculomotor tasks even in the human brainstem using conventional EPI imaging techniques. PMID- 21640193 TI - Evaluation of an algorithm for highly automated measurements of QT interval. AB - INTRODUCTION: Noise, artifact, and labile morphology of ECGs collected from freely moving animals in safety pharmacology studies render accurate measurements of QT interval challenging. Consequently, a high percentage of beats are uninterpretable and results provided by currently available analysis algorithms often require extensive manual review to correct errors. Performance of a novel algorithm, Multi-Domain Signal ProcessingTM (MDSP), is evaluated as a means of removing noise (denoising) without distorting morphology and for obtaining accurate beat-to-beat QT measurements. METHODS: Performance was evaluated using controlled experiments and an observational evaluation as follows: a) a clean ECG strip was intentionally corrupted with varying levels of noise to provide recordings of known signal-to-noise ratio (SNR). SNR and fidelity were compared pre- and post-MDSP denoising, b) beat-to-beat QT of a noisy ECG was measured manually pre- and post-MDSP denoising and automatically by MDSP, c) beat-to-beat QT of a clean ECG was measured manually and automatically using MDSP, and d) beat to-beat QT was computed for 3 freely moving non-human primates (NHP) pre- and post-torsadogen administration and the impact of averaging on QTSD, QT/RR dynamics and relationship was evaluated. RESULTS: MDSP reduced noise amplitude by up to 85% while preserving signal morphology. Mean QTs for manual and automatic measurements on a noisy ECG were within 2+/-15ms. MDSP-denoising prior to manual QT measurements resulted in a 22% decrease in QTSD compared to measurements obtained without denoising. Average QT standard deviation of the mean (QTSD) for automatic MDSP-derived measurements for 3 freely moving subjects was 7ms with 2.5% of beats automatically excluded due to noise. DISCUSSION: This work demonstrates that the MDSP algorithm shows promise as a tool for providing accurate automatic beat-to-beat measurements of QT interval from NHP in safety pharmacology studies. A methodology is presented for characterizing the impact of noise on algorithm performance. PMID- 21640194 TI - Optimization of a radiolabel DNA-binding assay in cultured mammalian cells. AB - An improved protocol for the radiolabel DNA-binding assay, which gives a high yield of highly pure DNA has been developed by use of mouse lymphoma cells. The critical difference from previously published methods is the use of enzymatic degradation of proteins in the later DNA purification steps rather than during the homogenisation procedure. Different DNA-purification methodologies were first compared and the protocol of choice was optimized later on; both steps were performed with [(35)S]-labelled amino acids for labelling of cellular protein, which enabled both the quantification of cellular protein contaminating the DNA sample and the distinction between cellular and enzyme-derived protein. The assay was later evaluated and shown to give reproducible results based on the data obtained with benzo[a]pyrene (B[a]P) and doxorubicin in two different laboratories. In addition, two further reference compounds, dopamine and diazepam and one proprietary AstraZeneca compound were also tested in mouse lymphoma cells in one laboratory. The two compounds B[a]P and doxorubicin were identified as suitable positive controls for routine testing in the presence and absence of S9, respectively. Exposing 90-100*10(6) cells to (14)C-labelled compound with a molar radioactivity of 2MBq/MUmol, yields approximately 500MUg DNA with <3% total protein contamination, of which approximately 7% is of cellular origin (<0.2%). The detection level is approximately 2adducts/10(8) dNTP. PMID- 21640195 TI - Genotoxic effects of doxorubicin in cultured human lymphocytes with different glutathione S-transferase genotypes. AB - Doxorubicin (Dox) is a widely used drug in oncology with a broad spectrum of interactions with various cellular components; therefore, it is likely to act through different mechanisms. In clinical practice there is inter-individual variability in cytotoxic drug response and in the occurrence of adverse reactions. Glutathione S-transferases (GSTM1, GSTT1 and GSTP1) are thought to be involved in the detoxification of endogenous and exogenous genotoxicants. The aim of this work is the assessment of a possible influence of polymorphisms in GSTs on the levels of genetic damage induced in vitro by Dox in cultured human lymphocytes. For this purpose, whole blood cultures from individuals with different genotypes for GSTM1, GSTT1 and GSTP1 were exposed to Dox and the cytokinesis-blocked micronucleus (CBMN) assay was used as the endpoint for chromosomal damage in the lymphocytes. Genotyping of GSTM1 and GSTT1 was carried out by multiplex PCR and the GSTP1-Ile105Val polymorphism was determined by PCR/RFLP. The total number of micronuclei present in 1000 binucleated cells and the frequency of micronucleated binucleated lymphocytes in the different individuals were analyzed considering the GSTM1, GSTT1 and GSTP1 genotypes. The results obtained suggest that GSTM1 and GSTT1 deletion polymorphisms do not modify significantly the genotoxic potential of Dox. However, the GSTP1 Ile105Val polymorphism was associated with an increase of micronucleated binucleated cells induced by Dox. Lymphocytes from homozygous individuals for the variant form (Val/Val) presented a significant increase in micronucleated binucleated cells (approximately 1.5-fold; p<0.05) when compared with individuals with at least one wild-type allele. These results suggest a possible role for GSTP1 on the modulation of the genotoxicity induced by Dox, which may be considered in cancer therapy. PMID- 21640196 TI - DNA-damage, cell-cycle arrest and apoptosis induced in BEAS-2B cells by 2 hydroxyethyl methacrylate (HEMA). AB - The methacrylate monomer 2-hydroxyethyl methacrylate (HEMA) is commonly used in resin-based dental restorative materials. These materials are cured in situ and HEMA and other monomers have been identified in ambient air during dental surgery. In vitro studies have demonstrated a toxic potential of methacrylates, and concerns have been raised regarding possible health effects due to inhalation. In this study we have investigated the mechanisms of HEMA-induced toxicity in the human lung epithelial cell line BEAS-2B. Depletion of cellular glutathione (GSH) and an increased level of reactive oxygen species (ROS) were seen after 2h of exposure, but the levels were restored to control levels after 12h. After 24h, inhibited cell proliferation and apoptotic cell death were found. The results of the Comet assay and the observed phosphorylation of DNA-damage associated signalling proteins including Chk2, H2AX, and p53 suggest that the toxicity of HEMA is mediated by DNA damage. Further, the antioxidant trolox did not counteract the HEMA-induced cell-cycle arrest, which indicates that the DNA damage is of non-oxidative origin. PMID- 21640197 TI - Seroprevalence of Toxoplasma gondii in wild boars (Sus scrofa leucomystax) and wild sika deer (Cervus nippon) in Gunma Prefecture, Japan. AB - The ingestion of undercooked meat from wild animals can be a source of Toxoplasma gondii infection in humans and other animals. In this study, we determined the seroprevalence of T. gondii infection in 175 wild boars (Sus scrofa leucomystax) and 107 wild sika deer (Cervus nippon) hunted in 2004-2007 in Gunma Prefecture, Japan, by using a commercial latex agglutination test (LAT). Antibodies (LAT, 1:64 or higher) to T. gondii were found in 6.3% of wild boars and 1.9% of sika deer. This is the first record of T. gondii infection in wild deer in Japan, and deer and wild boar meat should be cooked well before human consumption. PMID- 21640199 TI - Detection of the first G6P[14] human rotavirus strain from a child with diarrhea in Egypt. AB - We report the first detection of a G6P[14] rotavirus strain in Egypt from the stool of a child participating in a hospital-based diarrhea surveillance study conducted throughout the year 2004. Rotavirus infection was initially detected using a rotavirus group A VP6 enzyme immunoassay; the P (VP4) and G (VP7) genotypes of the strain were identified by RT-PCR. We sequenced the VP7 gene and the VP8* portion of the VP4 gene and the strain displayed the strongest identity to the VP7 [>94% nucleotides (nt), >97% amino acids (aa)] and VP4 (>93% nt, >98% aa) sequences of PA169, a novel G6P[14] strain first isolated from a child in Italy during the winter of 1987. Additional sequencing and analysis of the other remaining structural (VP1-VP3, VP6) and non-structural (NSP1-NSP5) proteins support this animal-to-human reassortment theory. According to the full genome classification system, the G6P[14] strain (EGY3399) was assigned to G6-P[14]-I2 R2-C2-M2-A11-N2-T6-E2-H3 genotypes. The greatest similarity of EGY3399 NSP4 and NSP5 gene sequences were to those of ovine and simian origin, respectively. Coupled with other observations, our results suggest G6P[14] isolates rarely cause severe diarrhea in Egyptian children, and support other studies that indicate animal rotavirus contribute to the genetic diversity of rotavirus detected from humans through interspecies transmission and single or multiple segments reassortment. PMID- 21640198 TI - Steatosis is an independent predictor of relapse following rapid virologic response in patients with HCV genotype 3. AB - BACKGROUND & AIMS: It is recommended that patients with chronic hepatitis C virus (HCV) genotype 3 infections receive 24 weeks of treatment. A rapid virologic response (RVR; at week 4) predicts a sustained virologic response (SVR), although not all patients with an RVR achieve an SVR. We explored the relationships among hepatic steatosis, level of HCV RNA, relapse, and RVR in a phase 3 randomized controlled trial of 932 patients infected with HCV genotype 2 (n = 427) or 3 (n = 505) who received 24 weeks of therapy with interferon-alpha. METHODS: In patients with an RVR (HCV RNA <43 IU/mL), the presence of an SVR was modeled using multivariate logistic regression as a function of age, sex, weight, body mass index, insulin resistance, steatosis, and levels of gamma-glutamyl transpeptidase, alanine aminotransferase, liver fibrosis, and baseline HCV RNA. RESULTS: RVR, SVR, and relapse rates among patients with HCV genotype 3 were 79.6%, 79.2%, and 15.6%, respectively; corresponding rates among patients with HCV genotype 2 were 86.7%, 84.3%, and 10.1%. An RVR had high predictive value for an SVR in patients with HCV genotypes 2 (88.9%) and 3 (88.1%). The strongest independent predictors of relapse in patients with genotype 3 and an RVR were steatosis (odds ratio 3.0; P = .003) and HCV RNA >=400,000 IU/mL (odds ratio 2.5; P = .04). Relapse rates in patients with steatosis were 17.4% and 20.9% for low and high baseline levels of HCV RNA, respectively; corresponding rates in those without steatosis were 2.5% and 8.8%. CONCLUSIONS: Steatosis was associated with significantly higher rates of relapse, irrespective of viral load, in patients infected with HCV genotype 3 who had an RVR. Further studies are needed to determine if longer treatment durations are effective in patients with an RVR and these risk factors. PMID- 21640200 TI - FK506 ameliorates renal injury in early experimental diabetic rats induced by streptozotocin. AB - Calcineurin (CaN) plays an important role in glomerular hypertrophy and extracellular matrix accumulation in early diabetic nephropathy. Cyclosporine (CSA), a CaN inhibitor, has been shown to reduce renal injury in streptozotocin induced diabetic rats. We examined whether FK506, which immunosuppressive action was 10-100 times of CSA, inhibits progression of diabetic nephropathy in experimental diabetic rats. Diabetes was induced with streptozotocin in rats, and FK506 (0.5 or 1.0mg/kg) was orally administered once a day for 4 weeks. Increased relative kidney weight was significantly reduced by FK506 treatment with 1.0mg/kg (p<0.05), and elevated 24 hour urinary albumin excretion rate was markedly attenuated by FK506 treatment with 0.5 and 1.0mg/kg (p<0.05, 0.01). Elevated glomerular volume was significantly attenuated by FK506 treatment with 0.5 and 1.0mg/kg (p<0.05), and increased indices for tubulointerstitial injury were only ameliorated by FK506 treatment with 1.0mg/kg (p<0.01). Western blot analysis noted that the expression of CaN protein was increased 2.4 fold in the kidney from diabetic rats, and FK506 treatment with 0.5 and 1.0mg/kg could reduce increased expression of CaN protein by 38.0% and 73.2%. The expression of 1alpha (IV) collagen, p65, p-p65, OPN, alpha-SMA and TGF-beta1 protein in kidney was significantly increased in diabetic rats and reduced by FK506 treatment (p<0.05, 0.01). Our results show that FK506 could ameliorate renal injury in early experimental diabetic rats, which mechanism may be at least partly correlated with suppression on increased CaN in renal tissue in diabetic rats. PMID- 21640201 TI - Naringin attenuates acute lung injury in LPS-treated mice by inhibiting NF-kappaB pathway. AB - Naringin has been reported as an effective anti-inflammatory compound. We previously showed that naringin had antitussive effect on experimentally induced cough in guinea pigs. However, the effects and mechanism of naringin on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice are not fully understood. In this study, our aim was to evaluate the anti-inflammatory activities of naringin on LPS-induced ALI in mice and clarify its underlying mechanisms of action. We found that in vivo pretreatment with naringin markedly decreased the lung wet weight to dry weight ratio, and led to significant attenuation of LPS-induced evident lung histopathological changes. Meanwhile, naringin significantly reduced bronchoalveolar lavage fluid (BALF) total cell and neutrophil (PMN) counts after LPS challenge. Furthermore, naringin inhibited myeloperoxidase (MPO: a marker enzyme of neutrophil granule) and inducible nitric oxide synthase (iNOS) activities in lung tissue and alleviated LPS-induced tumor neurosis factor-alpha (TNF-alpha) secretion in BALF in a dose-dependent manner. Additionally, Western blotting showed that naringin efficiently blunt NF-kappaB activation by inhibiting the degradation of IKB-alpha and the translocation of p65. Taken together, these results suggest that naringin shows anti-inflammatory effects through inhibiting lung edema, MPO and iNOS activities, TNF-alpha secretion and pulmonary neutrophil infiltration by blockade of NF-kappaB in LPS induced ALI. PMID- 21640202 TI - The power of life--cytochrome c oxidase takes center stage in metabolic control, cell signalling and survival. AB - Mitochondrial dysfunction is increasingly recognized as a major factor in the etiology and progression of numerous human diseases, such as (neuro )degeneration, ischemia reperfusion injury, cancer, and diabetes. Cytochrome c oxidase (COX) represents the rate-limiting enzyme of the mitochondrial respiratory chain and is thus predestined for being a central site of regulation of oxidative phosphorylation, proton pumping efficiency, ATP and reactive oxygen species production, which in turn affect cell signaling and survival. A unique feature of COX is its regulation by various factors and mechanisms interacting with the nucleus-encoded subunits, whose actual functions we are only beginning to understand. PMID- 21640203 TI - Proteases in autophagy. AB - Autophagy is a process involved in the proteolytic degradation of cellular macromolecules in lysosomes, which requires the activity of proteases, enzymes that hydrolyse peptide bonds and play a critical role in the initiation and execution of autophagy. Importantly, proteases also inhibit autophagy in certain cases. The initial steps of macroautophagy depend on the proteolytic processing of a particular protein, Atg8, by a cysteine protease, Atg4. This processing step is essential for conjugation of Atg8 with phosphatidylethanolamine and, subsequently, autophagosome formation. Lysosomal hydrolases, known as cathepsins, can be divided into several groups based on the catalitic residue in the active site, namely, cysteine, serine and aspartic cathepsins, which catalyse the cleavage of peptide bonds of autophagy substrates and, together with other factors, dispose of the autophagic flux. Whilst most cathepsins degrade autophagosomal content, some, such as cathepsin L, also degrade lysosomal membrane components, GABARAP-II and LC3-II. In contrast, cathepsin A, a serine protease, is involved in inhibition of chaperon-mediated autophagy through proteolytic processing of LAMP-2A. In addition, other families of calcium dependent non-lysosomal cysteine proteases, such as calpains, and cysteine aspartate-specific proteases, such as caspases, may cleave autophagy-related proteins, negatively influencing the execution of autophagic processes. Here we discuss the current state of knowledge concerning protein degradation by autophagy and outline the role of proteases in autophagic processes. This article is part of a Special Issue entitled: Proteolysis 50 years after the discovery of lysosome. PMID- 21640204 TI - Supercritical carbon dioxide processed resorbable polymer nanocomposite bone graft substitutes. AB - The development of synthetic bone graft substitutes is an intense area of research due to the complications associated with the harvest of autogenous bone and concerns about the supply of allogenic bone. Porous resorbable polymers have been used extensively in hard tissue engineering applications, but currently lack load-bearing capacity. Supercritical carbon dioxide (scCO(2)) processing is used as a novel method to simultaneously impart a porous structure and disperse a nano clay in a resorbable polymer matrix suitable for load-bearing applications. Porous resorbable polylactic acid (PLA)/cloisite clay nanocomposite constructs prepared using scCO(2) processing exhibit a 2.5-fold increase in compressive strength compared with pure polymer constructs. The resulting mechanical properties are comparable with human cancellous and cortico-cancellous bone. In addition to the significant improvements in mechanical properties, the nanocomposite constructs display a biocompatibility greater than that of polystyrene culture plate controls. Furthermore, calcium phosphate-rich deposits could clearly be seen on the surface of the constructs, as well as at the center of the cultured constructs, indicating that osteoblasts are able to penetrate the porous network of the nanocomposite constructs. Cellular infiltration of these constructs is important for their in vivo use as bone graft substitutes. The diameter of the pores suggests that these constructs would also support neovascularization, which is integral for nutrient transport. PMID- 21640205 TI - Perichondrium directed cartilage formation in silk fibroin and chitosan blend scaffolds for tracheal transplantation. AB - The purpose of this study was to investigate the potential of silk fibroin and chitosan blend (SFCS) biological scaffolds for the purpose of cartilage tissue engineering with applications in tracheal tissue reconstruction. The capability of these scaffolds as cell carrier systems for chondrocytes was determined in vitro and cartilage generation in vivo on engineered chondrocyte-scaffold constructs with and without a perichondrium wrapping was tested in an in vivo nude mouse model. SFCS scaffolds supported chondrocyte adhesion, proliferation, and differentiation, determined as features of the cells based on the spherical cell morphology, increased accumulation of glycosaminoglycans, and increased collagen type II deposition with time within the scaffold framework. Perichondrium wrapping significantly (P<0.001) improved chondrogenesis within the cell-scaffold constructs in vivo. In vivo implantation for 6weeks did not generate cartilage structures resembling native trachea, although cartilage-like structures were present. The mechanical properties of the regenerated tissue increased due to the deposition of chondrogenic matrix within the SFCS scaffold structural framework of the trachea. The support of chondrogenesis by the SFCS tubular scaffold construct resulted in a mechanically sound structure and thus is a step towards an engineered trachea that could potentially support the growth of an epithelial lining resulting in a tracheal transplant with properties resembling those of the fully functional native trachea. PMID- 21640206 TI - Existence of a typical threshold in the response of human mesenchymal stem cells to a peak and valley topography. AB - Our objective in this study was to determine whether a threshold in sensitivity of human mesenchymal stem cells (hMSC) to isotropic roughness exists. Using electrical discharge machining a very wide range of roughnesses (1.2MUm= 15 letters, proportion of patients with a loss of >15 letters, and safety. RESULTS: At week 12, treatment with VEGF Trap-Eye resulted in a significant mean decrease in CR/LT of 119 MUm from baseline in all groups combined (P<0.0001). The reduction in CR/LT with the 2 mg q4wk and 0.5mg q4wk regimens was significantly greater than each of the quarterly dosing regimens. The BCVA increased significantly by a mean of 5.7 letters at 12 weeks in the combined group (P<0.0001), with the greatest mean gain of >8 letters in the monthly dosing groups. At 8 weeks, BCVA improvements were similar with 2 mg q4wk and 2 mg q12wk dosing. After the last required dose at week 12, CR/LT and visual acuity were maintained or further improved at week 16 in all treatment groups. Ocular adverse events were mild and consistent with safety profiles reported for other intraocular anti-VEGF treatments. CONCLUSIONS: Repeated monthly intravitreal dosing of VEGF Trap-Eye over 12 weeks demonstrated significant reductions in retinal thickness and improvements in visual acuity, and was well-tolerated in patients with neovascular AMD. FINANCIAL DISCLOSURE(S): Proprietary or commercial disclosure may be found after the references. PMID- 21640258 TI - The 1-year results of CLEAR-IT 2, a phase 2 study of vascular endothelial growth factor trap-eye dosed as-needed after 12-week fixed dosing. AB - OBJECTIVE: To evaluate anatomic outcomes and vision, injection frequency, and safety during the as-needed (PRN) treatment phase of a study evaluating a 12-week fixed dosing period followed by PRN dosing to week 52 with vascular endothelial growth factor (VEGF) Trap-Eye for neovascular (wet) age-related macular degeneration (AMD). DESIGN: Multicenter, randomized, double-masked trial. PARTICIPANTS: We included 159 patients with subfoveal choroidal neovascularization (CNV) secondary to wet AMD. METHODS: Patients were randomly assigned to 1 of 5 intravitreal VEGF Trap-Eye treatment groups: 0.5 mg or 2 mg every 4 weeks or 0.5, 2, or 4 mg every 12 weeks during the fixed-dosing period (weeks 1-12). From weeks 16 to 52, patients were evaluated monthly and were retreated PRN with their assigned dose (0.5, 2, or 4 mg). MAIN OUTCOME MEASURES: Change in central retinal/lesion thickness (CR/LT), change in total lesion and CNV size, mean change in best-corrected visual acuity (BCVA), proportion of patients with 15-letter loss or gain, time to first PRN injection, reinjection frequency, and safety at week 52. RESULTS: The decrease in CR/LT at week 12 versus baseline remained significant at weeks 12 to 52 (-130 MUm from baseline at week 52) and CNV size regressed from baseline by 2.21 mm(2) at 48 weeks. After achieving a significant improvement in BCVA during the 12-week, fixed-dosing phase for all groups combined, PRN dosing for 40 weeks maintained improvements in BCVA to 52 weeks (5.3-letter gain; P<0.0001). The most robust improvements and consistent maintenance of visual acuity generally occurred in patients initially dosed with 2 mg every 4 weeks for 12 weeks, demonstrating a gain of 9 letters at 52 weeks. Overall, a mean of 2 injections was administered after the 12-week fixed-dosing phase, and the mean time to first reinjection was 129 days; 19% of patients received no injections and 45% received 1 or 2 injections. Treatment with VEGF Trap-Eye was generally safe and well tolerated, with few ocular or systemic adverse events. CONCLUSIONS: PRN dosing with VEGF Trap-Eye at weeks 16 52 maintained the significant anatomic and vision improvements established during the 12-week fixed-dosing phase with a low frequency of reinjections. Repeated dosing with VEGF Trap-Eye was well tolerated over 52 weeks of treatment. FINANCIAL DISCLOSURE(S): Proprietary or commercial disclosure may be found after the references. PMID- 21640259 TI - Early resident-performed cataract surgery. PMID- 21640260 TI - Phacotrabeculectomy meta-analysis. PMID- 21640261 TI - Trends and impact of ophthalmology research. PMID- 21640262 TI - Endothelin-1 in neovascular AMD. PMID- 21640263 TI - Pediatric IOL calculation. PMID- 21640265 TI - Pediatric IOL calculation. PMID- 21640267 TI - Intraocular lens calculations. PMID- 21640269 TI - Preschool amblyopia screening. PMID- 21640271 TI - Corneal dystrophy recurrence. PMID- 21640272 TI - Microperimeter MP1. PMID- 21640274 TI - Body mass, spinal fluid, and glaucoma. PMID- 21640275 TI - Body mass, spinal fluid, and glaucoma. PMID- 21640277 TI - [Introduction. Distal airways in asthma and COPD]. PMID- 21640278 TI - [Inflammation and remodeling of the distal airways: studies in humans and experimental models]. AB - Asthma is characterized by inflammation and remodeling of the airways, giving rise to airway obstruction and symptoms of wheezing, chest tightness, cough and dyspnea. Most of these observations arise from the study of samples obtained from the central airways by distinct methods. However, it is currently accepted that this inflammatory process occurs not only in the central airway but also in the small airway and even in the pulmonary parenchyma of all asthmatic patients, even those with mild asthma. CD4+ lymphocytes, activated eosinophils and IL-5 mRNA expression are present in a greater quantity in the small airways. Also present is remodeling, with an increase in submucosal thickness, the muscular layer and adventitia. This inflammatory process causes a disconnection between the pulmonary parenchyma and the airway, giving rise to obstruction of the small airway, which is currently considered to be predominant in asthmatic patients. Likewise, studies of experimental asthma in animals support the substantial role of the distal airway. Recognition that asthma affects the entire airway could be clinically important and lead to the distal lung being considered as a target in any effective therapeutic strategy. However, longitudinal studies are required to evaluate the impact of distal airway inflammation and its treatment in asthma. PMID- 21640279 TI - [Physiology and physiopathology of the distal airways in asthma]. AB - The small airways are those with an internal diameter of less than 2 mm. The contribution of these airways to total airflow resistance is small in healthy individuals but can represent 50-90 % of total airflow resistance in asthmatics. Suspicion of small airways disease has been based on reduction of midexpiratory and instantaneous flows, although wide variability in their values and the absence of a sufficiently validated cut-off point has limited their clinical application. Static pulmonary volumes can provide indirect evidence of the state of the most distal airways, revealing two effects of their alteration: air trapping and dynamic hyperinflation. While determination of airway resistance by plethysmography and of respiratory system resistance measured by flow interruption are highly non-specific, the forced oscillation technique allows obstruction of the small airways to be distinguished from that of medium-caliber airways. The characteristic pattern of peripheral obstruction includes a decrease in frequency-dependent resistance, reduced reactivity and an increase in resonance frequency. Single-or multiple-breath nitrogen washout can also provide specific information on the small airways, although the apparatus required is less frequently available. Analysis through bicompartmental models of exhaled nitric oxide allows alveolar nitric oxide concentrations to be determined, which seems to provide information on inflammatory activity in the small airways. PMID- 21640280 TI - [Clinical relevance of distal airway involvement in asthma]. AB - Asthma continues to be a global health problem, despite advances in diagnostic techniques and treatment. The inflammatory nature of asthma is currently indisputable, as is the involvement of the entire respiratory tree, both the proximal and most distal airways, which has been demonstrated in multiple studies. The development of the therapeutic arsenal, with more potent drugs and improved inhalation devices, has allowed a certain control to be maintained over the inflammatory process, although the inability to reach the most distal points of the airways has posed a stumbling block that seems difficult to overcome. However, the available information on the real role of distal airway involvement in asthma remains very scarce. Physiopathological evidence shows that, in addition to the large airways, the small or distal airways (those with a diameter of less than 2 mm) substantially contribute to the severity of asthma. Several studies have shown that the inflammatory process seems to be more intense in this area. This finding has been related to nocturnal asthma and an increase in glucocorticoid receptor-beta-expressing cells, associated with corticosteroid resistant asthma and fatal asthma. Equally, small airway involvement seems to be a highly important factor in asthma in the pediatric age group. PMID- 21640281 TI - [Imaging techniques in the examination of the distal airways: asthma and COPD]. AB - Imaging techniques are highly useful diagnostic tools to study small airway diseases. Despite their differences, from a clinical pathological perspective, these diseases show similar radiological manifestations. High-resolution computed tomography (CT) is the technique of choice to study diffuse diseases and those of the small airways; the slices obtained in expiratory high-resolution CT scan should form part of the study protocol of small airway diseases. Based on the findings of high-resolution CT, small airway diseases can be divided into two large groups: (a) those presenting direct morphological signs of bronchiolar involvement, and (b) those showing indirect signs of bronchiolar involvement (air trapping/mosaic pattern). High-resolution CT is highly useful to study the complications of asthma (allergic bronchopulmonary aspergillosis) and to evaluate clinically similar processes, such as hypersensitivity pneumonitis. In asthmatic patients, multi-detector CT (MDTC) allows clinical symptoms, thickening of the airway wall and the degree of airflow obstruction to be directly correlated. MDTC is also useful for quantitative evaluation of the degree of air trapping in patients with emphysema. Magnetic resonance imaging after inhalation of distinct gases, such as (3)He and xenon-129 and dual-energy CT are also useful imaging techniques in the direct or indirect evaluation of the degree of airflow obstruction (air trapping). PMID- 21640282 TI - [Therapeutic approach to the distal airways in asthma]. AB - Bronchial asthma is a chronic inflammatory disease that affects both the large caliber airways and the smaller-caliber bronchioles. In the last few years, a major therapeutic advance has been made with the development of new systems of inhalation solutions, which produce extra-fine particles, achieving better lung deposition throughout the airways and reducing oropharyngeal deposition. These formulations have improved the effectiveness of bronchodilation and particularly the antiinflammatory effect. The use of long-acting b2-adrenergic steroids in extra-fine formulation, whether alone or in combinations of fixed doses, improves drug distribution throughout the bronchial tree, enhancing the therapeutic effect with lower doses of drugs. Leukotriene receptor antagonists have shown their effect on the small airways in asthmatic patients, both in studies of pulmonary resistance and pulmonary volumes and in imaging studies. PMID- 21640283 TI - [Importance of the distal airway in COPD]. AB - Chronic obstructive pulmonary disease (COPD) is characterized by inflammation causing airflow obstruction. However, the initial histological lesion of COPD occurs in the respiratory bronchiole and spirometry is unable to detect involvement of this area until the disease is advanced. Major advances have been made in characterizing the inflammatory process in the small airways. However, in clinical practice, a non-invasive marker of small airways involvement, which would allow injury and the effect of treatment to be monitored, is lacking. To date, the combination of bronchodilators and inhaled corticosteroids is recommended for the most severe cases, although the effects of this therapeutic option on the small airways are not well known. New treatments that reach the distal airways and novel techniques to assess the small airways will allow a more complete approach to this disease. PMID- 21640284 TI - [Introduction. Thoracic surgery]. PMID- 21640285 TI - [Preoperative estimation of the risk of lung resection]. AB - The present article discusses the two most up-to-date clinical practice guidelines containing the recommendations of US and European scientific societies on preoperative assessment of the risk of lung resection. Despite some differences between the two documents, both guidelines agree on the importance of routine preoperative measurement of diffusion lung capacity for carbon monoxide (DLCO) in the predictive value of exercise tests, especially measurement of maximal oxygen uptake per minute (VO(2max)). Precisely because of its ability to predict the risk of operative death, VO(2max) should be measured in patients with a forced expiratory volume in 1 second (FEV1) or DLCO below 80% of the theoretical value. The authors recommend using one of the two above-mentioned guidelines in clinical practice and periodically auditing the results to compare them with in-hospital mortality for lung resection in Europe, currently available through the European Association of Thoracic Surgeons. There is currently no validated risk index that could be directly applied in clinical decision making in lung resection. PMID- 21640286 TI - [Surgery of lung metastases]. AB - The aim of surgical treatment of lung metastases is to eliminate all known tumoral disease. After a clinical diagnosis of lung metastases, the criteria for selecting patients who are candidates for surgical treatment, the route of access to the thoracic cavity and the technique for metastases resection are not universally defined. Moreover, half of all patients will show recurrence and the advisability of further surgery will have to be reconsidered. The present article discusses aspects related to the oncological and functional limits of surgical resection of lung metastases, preoperative workup, postoperative follow-up, and the surgical approaches and resection techniques. PMID- 21640287 TI - [Chest trauma]. AB - Chest trauma is a frequent problem arising from lesions caused by domestic and occupational activities and especially road traffic accidents. These injuries can be analyzed from distinct points of view, ranging from consideration of the most severe injuries, especially in the context of multiple trauma, to the specific characteristics of blunt and open trauma. In the present article, these injuries are discussed according to the involvement of the various thoracic structures. Rib fractures are the most frequent chest injuries and their diagnosis and treatment is straightforward, although these injuries can be severe if more than three ribs are affected and when there is major associated morbidity. Lung contusion is the most common visceral lesion. These injuries are usually found in severe chest trauma and are often associated with other thoracic and intrathoracic lesions. Treatment is based on general support measures. Pleural complications, such as hemothorax and pneumothorax, are also frequent. Their diagnosis is also straightforward and treatment is based on pleural drainage. This article also analyzes other complex situations, notably airway trauma, which is usually very severe in blunt chest trauma and less severe and even suitable for conservative treatment in iatrogenic injury due to tracheal intubation. Rupture of the diaphragm usually causes a diaphragmatic hernia. Treatment is always surgical. Myocardial contusions should be suspected in anterior chest trauma and in sternal fractures. Treatment is conservative. Other chest injuries, such as those of the great thoracic and esophageal vessels, are less frequent but are especially severe. PMID- 21640288 TI - [Chest wall surgery]. AB - Despite the numerous differences among the distinct diseases of the chest wall, the surgery of this area shows certain common features. Treatment has progressively changed in the last few years due to advances in diagnostic techniques, minimally invasive procedures and reconstruction materials, and especially due to the multidisciplinary management of many diseases. Nuss' minimally invasive correction of pectus excavatum has gained devotees, although open approaches are performed with increasingly small incisions, almost comparable to the lateral incisions in Nuss' technique. Surgeons supporting the open approach also cite the evident disadvantages of the need for a steel implant for 2 or 3 years and for a second intervention to remove this implant. En-bloc resections with reconstruction using materials, which are increasingly better and covered by myocutaneous grafts in collaboration with plastic surgery departments, constitute a major advance in the treatment of chest wall tumors. Trimodal therapy for Pancoast tumors, consisting of induction chemotherapy and radiotherapy and subsequent surgical treatment of the tumor, currently provides the best results in terms of resectability and survival. PMID- 21640289 TI - [Summary of the clinical studies reported in the Annual Scientific Sessions of the American College of Cardiology (New Orleans, LA, USA, April 2-5, 2011)]. PMID- 21640290 TI - The use of antiplatelet therapy in the outpatient setting: Canadian Cardiovascular Society guidelines. AB - Antiplatelet agents are a cornerstone of therapy for patients with atherosclerotic vascular disease. There is presently a lack of comprehensive guidelines focusing on the use of antiplatelet drugs in patients currently manifesting or at elevated risk of cardiovascular disease. The Canadian Antiplatelet Therapy Guidelines Committee reviewed existing disease-based guidelines and subsequently published literature and used expert opinion and review to develop guidelines on the use of antiplatelet therapy in the outpatient setting. This full document has been summarized in an Executive Summary published in the Canadian Journal of Cardiology and may be found at http://www.ccs.ca/. Antiplatelet therapy appears to be generally underused, perhaps in part because of a lack of clear, evidence-based guidance. Here, we provide specific guidelines for secondary prevention in patients discharged from hospital following acute coronary syndromes, post-percutaneous coronary intervention, post-coronary artery bypass grafting, patients with a history of transient cerebral ischemic events or strokes, and patients with peripheral arterial disease. Issues related to primary prevention are also addressed, in addition to special clinical contexts such as diabetes, heart failure, chronic kidney disease, pregnancy/lactation, and perioperative management. Recommendations are provided regarding pharmacologic interactions that may occur during combination therapy with warfarin, clopidogrel and proton-pump inhibitors, or acetylsalicylic acid and nonsteroidal anti inflammatory drugs, as well as for the management of bleeding complications. PMID- 21640294 TI - Imaging acute stroke is rapidly evolving. PMID- 21640292 TI - Estimated effect of prenatal cocaine exposure on examiner-rated behavior at age 7 years. AB - Prenatal cocaine exposure has been linked to increased child behavior difficulties in some studies but not others. OBJECTIVE: The primary aim was to estimate the relationship between in utero cocaine exposure and child behavioral functioning at age 7 years with ratings made by blinded examiners during a structured testing session. A second aim was to examine whether caregiver drug use and psychological problems might mediate suspected relationships between prenatal cocaine exposure and aspects of examiner-rated behavior. METHODS: 407 children (212 cocaine-exposed, 195 non-exposed) participating in the longitudinal Miami Prenatal Cocaine Study (MPCS) were rated with regard to their behavior during a neuropsychological assessment conducted at age 7 years. Raters were trained research psychometricians blinded to drug exposure status. Individual behavioral items were summarized and the cocaine-behavior relationship was estimated within the context of latent variable modeling, using Mplus software. RESULTS: Two latent variables, Behavioral Regulation and Sociability, were derived via exploratory latent structure analysis with promax rotation. Prenatal cocaine exposure, statistically controlling for child sex, test age, and prenatal exposure to alcohol, tobacco, and marijuana, was associated with Behavioral Regulation (estimated slope beta=-0.25; 95% CI=-0.48, -0.02; p=0.04) but not Sociability (estimated slope beta=-0.03; 95% CI=-0.26, 0.20; p=0.79). Neither postnatal drug use by caregivers nor the severity of their psychological problems at age 5 follow-up predicted levels of child Behavioral Regulation or Sociability at age 7 years (p>0.10). CONCLUSIONS: Examiner ratings of child behavior at age 7 revealed less optimal behavioral regulation for prenatally cocaine-exposed compared to non-exposed children, in contrast with what had been previously found from parent-report data. This evidence highlights the potential value of trained observers in assessing behavioral outcomes of children exposed in utero to drugs and other toxicants. PMID- 21640295 TI - Advanced CT imaging in the evaluation of hemorrhagic stroke. AB - Multidetector computed tomographic (CT) angiography is rapidly becoming a pivotal examination in the initial evaluation of patients with hemorrhagic stroke. This article provides an update of the literature on this dynamic topic, focusing on (1) the utility of CT angiography in the identification of hemorrhagic stroke patients who harbor an underlying vascular etiology and the role of the secondary intracerebral hemorrhage score, as well as (2) the clinical value of the CT angiography spot sign and spot sign score in patients with primary intracerebral hemorrhage. PMID- 21640296 TI - CT perfusion imaging in acute stroke. AB - Computed tomographic perfusion (CTP) imaging is an advanced modality that provides important information about capillary-level hemodynamics of the brain parenchyma. CTP can aid in diagnosis, management, and prognosis of acute stroke patients by clarifying acute cerebral physiology and hemodynamic status, including distinguishing severely hypoperfused but potentially salvageable tissue from both tissue likely to be irreversibly infarcted ("core") and hypoperfused but metabolically stable tissue ("benign oligemia"). A qualitative estimate of the presence and degree of ischemia is typically required for guiding clinical management. Radiation dose issues with CTP imaging, a topic of much current concern, are also addressed in this review. PMID- 21640297 TI - Stroke imaging research road map. AB - Although acute stroke imaging has made significant progress in the last few years, several improvements and validation steps are needed to make stroke imaging techniques fully operational and appropriate in daily clinical practice. This review outlines the needs in the stroke-imaging field and describes a consortium that was founded to provide them. PMID- 21640298 TI - Role of diffusion and perfusion MRI in selecting patients for reperfusion therapies. AB - After onset of ischemic stroke, potentially viable tissue at risk (ischemic penumbra) may be salvageable. Currently, intravenous alteplase is approved for up to 4.5 hours after symptom onset of acute ischemic stroke. Increasing this time window may allow many more patients to be treated. The ability to use MRI to help define the irreversibly damaged brain (infarct core) and the reversible ischemic penumbra shows great promise for stroke treatment. Recent advances in penumbral imaging technology may enable a phase III trial of an intravenous thrombolytic to be performed beyond 4.5 hours using techniques to select patients with penumbral tissue. PMID- 21640299 TI - MR perfusion imaging in acute ischemic stroke. AB - Magnetic resonance (MR) perfusion imaging offers the potential for measuring brain perfusion in acute stroke patients, at a time when treatment decisions based on these measurements may affect outcomes dramatically. Rapid advancements in both acute stroke therapy and perfusion imaging techniques have resulted in continuing redefinition of the role that perfusion imaging should play in patient management. This review discusses the basic pathophysiology of acute stroke, the utility of different kinds of perfusion images, and research on the continually evolving role of MR perfusion imaging in acute stroke care. PMID- 21640300 TI - Arterial spin label imaging of acute ischemic stroke and transient ischemic attack. AB - Since acute stroke and transient ischemic attack (TIA) are disruptions of brain hemodynamics, perfusion neuroimaging might be of clinical utility. Recently, arterial spin labeling (ASL), a noncontrast perfusion method, has become clinically feasible. It has advantages compared to contrast bolus perfusion weighted imaging (PWI) including lack of exposure to gadolinium, improved quantitation, and decreased sensitivity to susceptibility and motion. Drawbacks include reduced signal-to-noise and high sensitivity to arterial transit delays. However, this sensitivity can enable visualization of collateral flow. This article discusses ASL findings in patients with acute stroke and TIA, focusing on typical appearances, common artifacts, and comparisons with PWI. PMID- 21640301 TI - Transient ischemic attack: definition, diagnosis, and risk stratification. AB - Transient ischemic attack (TIA) can convey a high imminent risk for the development of a major stroke and is therefore considered to be a medical emergency. Recent evidence indicates that TIA with imaging proof of brain infarction represents an extremely unstable condition with early risk of stroke that is as much as 20 times higher than the risk after TIA without tissue damage. The use of neuroimaging in TIA is therefore critical not only for diagnosis but also for accurate risk stratification. In this article, recent advances in diagnostic imaging, categorizations, and risk stratification in TIA are discussed. PMID- 21640302 TI - Measuring permeability in acute ischemic stroke. AB - The blood-brain barrier (BBB) is a functional concept to describe unique features of intracranial blood vessels that prevent many substances in the systemic circulation from entering the brain. In the setting of acute ischemic stroke, loss of blood-brain barrier (BBB) integrity is believed to be a precursor to hemorrhagic transformation. CT and MR imaging may evaluate BBB integrity by detecting leakage of intravenously administered contrast media into the extravascular space. In its simplest form, BBB integrity is assessed qualitatively, by determining the presence or absence of contrast enhancement on structural images of the brain. When dynamic contrast-enhanced (DCE) MRI or CT is combined with a suitable pharmacokinetic model, one can quantify and spatially map BBB integrity throughout the brain. PMID- 21640304 TI - MR diffusion imaging in ischemic stroke. AB - Diffusion-weighted MRI provides image contrast that is dependent on the molecular motion of water. Diffusion-weighted imaging is the most reliable method for early detection of cerebral ischemia, for the definition of infarct core, and for the differentiation of acute ischemia from other disease processes that mimic stroke. Diffusion tensor imaging and diffusion kurtosis imaging may offer additional diagnostic information on the microstructural status of tissue. This review discusses the development and applications of diffusion-weighted imaging, diffusion tensor imaging, and diffusion kurtosis imaging in acute and chronic ischemia. PMID- 21640303 TI - Imaging stroke patients with unclear onset times. AB - Stroke is a leading cause of death and adult morbidity worldwide. By defining stroke symptom onset by the time the patient was last known to be well, many patients whose onsets are unwitnessed are automatically ineligible for thrombolytic therapy. Advanced brain imaging may serve as a substitute witness to estimate stroke onset and duration in those patients who do not have a human witness. This article reviews and compares some of these imaging-based approaches to thrombolysis eligibility, which can potentially expand the use of thrombolytic therapy to a broader population of acute stroke patients. PMID- 21640305 TI - Intra-arterial stroke therapy: recanalization strategies, patient selection and imaging. AB - With more than 700,000 strokes per year resulting in greater than 160,000 deaths per year, stroke remains the leading cause of disability and third leading cause of death in the United States. Despite an overall decline in stroke mortality over the past 40 years, the total number of stroke deaths continues to increase, suggesting an increase in stroke incidence. The last 20 years of neuroscience advances have moved stroke from a condition that is monitored clinically and imaged serially as it evolves to an entity that can be treated acutely, with remarkable alterations in its natural history. PMID- 21640306 TI - Noninvasive carotid artery imaging with a focus on the vulnerable plaque. AB - Currently carotid imaging has 2 main focuses: assessment of luminal stenosis and classification of atherosclerotic plaque characteristics. Measurement of the degree of stenosis is the main assessment used for current treatment decision making, but an evolving idea that is now driving imaging is the concept of vulnerable plaque, which is where plaque components are identified and used to define which plaques are at high risk of causing symptoms compared with those at low risk. This review article covers the methods used for noninvasive assessment of carotid luminal stenosis and the options available for plaque imaging. PMID- 21640307 TI - ASPECTS and other neuroimaging scores in the triage and prediction of outcome in acute stroke patients. AB - Information obtained from brain imaging is now summarized in the form of various neuroimaging scores to help physicians in making therapeutic decisions and determining prognosis. The Alberta Stroke Program Early CT Score (ASPECTS) was devised to quantify the extent of early ischemic changes in the middle cerebral artery territory on noncontrast computed tomography. With its systematic approach, the score is simple, reliable, and a strong predictor of functional outcome. This review summarizes ASPECTS and other neuroimaging scores developed for risk prognostication and risk stratification with treatment in patients with acute ischemic stroke. PMID- 21640308 TI - In-vitro culture of human embryos with mechanical micro-vibration increases implantation rates. AB - The in-vitro culture of human embryos in a medium subjected to regular short intervals of mechanical agitation leads to increased development rates. This type of treatment tries to mimic conditions in nature whereby oviductal fluid is mechanically agitated by the epithelial cilia. This phenomenon can be explained by the fact that an embryo developing in vivo is naturally exposed to constant vibrations of around 6Hz with the periodically repeating increase to 20Hz. This review covers the history of this question and in this light offers an explanation through biological concept for one of the most recent developments in this area: in-vitro culture of human embryos with mechanical micro-vibration. The effect of mechanical micro-vibration on embryos during their in-vitro culture was examined. Pregnancy rates after the transfer of embryos in the group with in vitro culture under mechanical vibration were increased. PMID- 21640309 TI - Early pregnancy wastage: ethical considerations. AB - Information on early embryo wastage is relevant for debating the status of human embryos. Two main points of view confront each other. Theists hold that human embryos should be treated as human persons from the moment of conception because, even accepting that human beings are the fruit of evolution, they are part of a divine project. Without a developmental event prior to which the human embryo could not be considered a human being, embryos should be regarded as if they were human subjects. After all, if one believes in the resurrection of the dead, it makes no difference at what stage one's life ends. Secularists oppose the idea of granting absolute value to human life from its beginning because early human embryos lack individuality and sentience. Personifying embryos is morally absurd because it would mean that countless human beings never had even the slightest chance to express their potential and, in the light of this catastrophic loss, one would expect early pregnancy wastage to have become an important research priority; this is not the case. In practical terms, most Western countries have legalized first-trimester abortion, de facto giving embryos a lower status than that of full person. PMID- 21640310 TI - Introduction: The 200th anniversary of the hypha. PMID- 21640311 TI - Architecture and development of the Neurospora crassa hypha -- a model cell for polarized growth. AB - Neurospora crassa has been at the forefront of biological research from the early days of biochemical genetics to current progress being made in understanding gene and genetic network function. Here, we discuss recent developments in analysis of the fundamental form of fungal growth, development and proliferation -- the hypha. Understanding the establishment and maintenance of polarity, hyphal elongation, septation, branching and differentiation are at the core of current research. The advances in the identification and functional dissection of regulatory as well as structural components of the hypha provide an expanding basis for elucidation of fundamental attributes of the fungal cell. The availability and continuous development of various molecular and microscopic tools, as utilized by an active and co-supportive research community, promises to yield additional important new discoveries on the biology of fungi. PMID- 21640312 TI - Hyphal morphogenesis: an evolutionary perspective. AB - Two modes of cellular morphogenesis predominate within the fungal kingdom; yeast growth and hyphal growth. The availability of complete genome sequences that span the kingdom has made possible the use of comparative approaches that address important questions regarding the evolution of these growth modes. These comparisons have also emphasized the point that not all hyphae are the same despite outward appearances. Topics considered here include the origins of hyphal growth, as well as the potential causes of and the consequences resulting from the loss of hyphal growth in yeast lineages. The mechanisms that enable distinct morphological outputs (i.e., yeast vs. hyphae) using an essentially identical inventory of gene products are also considered. Finally, processes implicated in the regulation of hyphal tip complexes are addressed from an evolutionary perspective. PMID- 21640313 TI - The hyphal tip structure of Basidiobolus sp.: a zygomycete fungus of uncertain phylogeny. AB - To date, among the zygomycete fungi that have been examined, a Spitzenkorper has not been reported. In this paper, the cytoplasmic order of hyphal tip cells of Basidiobolus sp., a zygomycete genus of uncertain phylogeny, has been examined using light microscopy and transmission electron microscopy methods. With phase contrast light optics, a phase-dark body was observed at the tips of growing hyphae of Basidiobolus sp. The hyphal apex also showed high affinity for FM4-64 labelling resulting in an intense fluorescence signal. The phase-dark inclusion exhibited independent motility within the hyphal apex and its presence and position were correlated to the rate and direction of hyphal growth. The hyphal apex of Basidiobolus sp. did not contain gamma-tubulin. Ultrastructural observations revealed a dense cluster of vesicles at the hyphal apex. These results suggest that the growing hypha of Basidiobolus sp. contains a Spitzenkorper, a character generally attributed to members of the ascomycete and basidiomycete fungi and not to zygomycete fungi. PMID- 21640314 TI - Probing the growth dynamics of Neurospora crassa with microfluidic structures. AB - Despite occupying physically and chemically heterogeneous natural environments, the growth dynamics of filamentous fungi is typically studied on the surface of homogeneous laboratory media. Fungal exploration and exploitation of complex natural environments requires optimal survival and growth strategies at the colony, hyphal, and intra hyphal level, with hyphal space-searching strategies playing a central role. We describe a new methodology for the characterisation and analysis of hyphal space-searching strategies, which uses purposefully designed three-dimensional microfluidics structures mimicking some of the characteristics of natural environments of the fungi. We also demonstrate this new methodology by running a comparative examination of two Neurospora crassa strains, i.e., the wild type of N. crassa -- a commonly used model organism for the study of filamentous fungi -- and the N. crassa ro-1 mutant strain -- which is deficient in hyphal and mycelial growth. Continuous live imaging showed that both strains responded actively to the geometrically confined microstructured environments without any detectable temporal delay or spatial adjustment. While both strains navigated the test structures exhibiting similar geometry-induced space-searching mechanisms, they presented fundamentally different growth patterns that could not be observed on geometrically unconfined, flat agar surfaces. PMID- 21640315 TI - On the role of microtubules, cell end markers, and septal microtubule organizing centres on site selection for polar growth in Aspergillus nidulans. AB - Tip growth of filamentous fungi depends on continuous polarized growth and requires the actin and microtubule (MT) cytoskeleton. Cortical proteins at polarity sites, also known as cell end markers, play important roles in polarity maintenance. Deletion of the cell end marker teaA results in zigzag hyphal morphologies, which is contrary to the normal rectilinear growth pattern. Here we studied the role of TeaA and MTs in the establishment of polarity during tip growth of Aspergillus nidulans, including conidia germination, second germtube formation, hyphal branching and conidiophore development. TeaA is delivered to the cortex by growing MTs. In conidia TeaA appeared at the germination site prior to germtube formation, and deletion of teaA resulted in germination at multiple sites, increased branching and abnormal conidiophores. The formation of a second germtube opposite the first conidial germtube depended on the presence of a septum at the base of the first germtube. An MT-organizing centre, associated to the septum, produced microtubules, which delivered TeaA towards the opposite side of the conidium. These results suggest a new function for TeaA in polarity establishment. It can be a positive function, but TeaA could also suppress polarity sites in the vicinity of the first germtube. PMID- 21640316 TI - Form follows function -- the versatile fungal cytoskeleton. AB - The cytoskeleton plays a major role in the regulation of fungal cell morphogenesis. The fungal cytoskeleton is comprised of three polymers: F-actin, microtubules and septins. Due to the successful application of the newly developed Lifeact probe for live-cell imaging of F-actin it is now possible, in combination with existing microtubule markers and fluorescently labelled septins, to monitor real-time dynamics of the entire fungal cytoskeleton, and reassess the many and integrated roles of F-actin, microtubules and septins throughout fungal growth and development. Evidence is accumulating that functional properties of higher-order structures derived from actin and septin filaments interacting with microtubules are employed in different ways in different cell types. This may reflect marked differences in cytoskeletal architecture that are found, for example, in unicellular yeasts, spore germlings and mature fungal hyphae. In this review we address key aspects of the versatile fungal cytoskeleton, highlight recently gained insights into important roles of F-actin in filamentous fungi, and raise some key questions that are likely to be solved in the coming years based on the new experimental tools that have recently become available. PMID- 21640317 TI - A role for endocytic recycling in hyphal growth. AB - Actin plays multiple complex roles in cell growth and cell shape. Recently it was demonstrated that actin patches, which represent sites of endocytosis, are present in a sub-apical collar at growing tips of hyphae and germ tubes of filamentous fungi. It is now clear that this zone of endocytosis is necessary for filamentous growth to proceed. In this review evidence for the role of these endocytic sites in hyphal growth is examined. One possibility if that the role of the sub-apical collar is associated with endocytic recycling of polarized material at the hyphal tip. The 'Apical Recycling Model' accounts for this role and predicts the need for a balance between endocytosis and exocytosis at the hyphal tip to control growth and cell shape. Other cell differentiation events, including appressorium formation and Aspergillus conidiophore development may also be explained by this model. PMID- 21640318 TI - The mating projections of Saccharomyces cerevisiae and Candida albicans show key characteristics of hyphal growth. AB - Fungi can grow in a variety of growth forms: yeast, pseudohyphae and hyphae. The human fungal pathogen Candida albicans can grow in all three of these forms. In this fungus, hyphal growth is distinguished by the presence of a Spitzenkorper like structure at the hyphal tip and a band of septin bars around the base of newly evaginated germ tubes. The budding yeast Saccharomyces cerevisiae grows as yeast and pseudohyphae, but is not normally considered to show hyphal growth. We show here that in mating projections of both C. albicans and S. cerevisiae a Spitzenkorper-like structure is present at the growing tip and a band of septin bars is present at the base. Furthermore, in S. cerevisiae mating projections, Spa2 and Bni1 form a cap to the 3-dimensional ball of FM4-64 staining, exactly as previously observed in C. albicans hyphae, suggesting that the putative Spitzenkorper may be a distinct structure from the polarisome. Taken together this work shows that mating projections of both S. cerevisiae and C. albicans show the key characteristics of hyphal growth. PMID- 21640319 TI - Evolution of multinucleated Ashbya gossypii hyphae from a budding yeast-like ancestor. AB - In the filamentous ascomycete Ashbya gossypii polarity establishment at sites of germ tube and lateral branch emergence depends on homologues of Saccharomyces cerevisiae factors controlling bud site selection and bud emergence. Maintenance of polar growth involves homologues of well-known polarity factors of budding yeast. To achieve the much higher rates of sustained polar surface expansion of hyphae compared to mainly non-polarly growing yeast buds five important alterations had to evolve. Permanent presence of the polarity machinery at a confined area in the rapidly expanding hyphal tip, increased cytoplasmic space with a much enlarged ER surface for generating secretory vesicles, efficient directed transport of secretory vesicles to and accumulation at the tip, increased capacity of the exocytosis system to process these vesicles, and an efficient endocytosis system for membrane and polarity factor recycling adjacent to the zone of exocytosis. Morphological, cell biological, and molecular aspects of this evolution are discussed based on experiments performed within the past 10 y. PMID- 21640320 TI - Coalescence map for bubbles in surfactant-free aqueous electrolyte solutions. AB - Factors influencing bubble coalescence in surfactant-free aqueous electrolyte solutions are considered in this compilation of literature results. These factors include viscous and inertial thin film drainage, surface deformation, surface elasticity, mobility or otherwise of the air-water interface, and disjoining pressure. Several models from the literature are discussed, with particular attention paid to predictions of transitions between regions where behaviour is qualitatively different. The transitions are collated onto a single chart with salt concentration and bubble approach speed as the axes. This creates a map of the regions in which different mechanisms operate, giving an overall picture of bubble coalescence behaviour over a wide range of concentration and speed. Only mm-size bubbles in water and NaCl solutions are discussed in this initial effort at creating such a map. Data on bubble coalescence or non-coalescence are collected from the literature and plotted on the same map, generally aligning well with the predicted transitions and thus providing support for the theoretical reasoning that went into creating the coalescence map. PMID- 21640321 TI - Effectiveness of implantable defibrillators in octogenarians and nonagenarians for primary prevention of sudden cardiac death. AB - Given their advanced age and frequent co-morbidities, it is unclear whether octogenarians and nonagenarians with decreased left ventricular ejection fraction (LVEF) derive a survival benefit from implantable cardioverter-defibrillators (ICDs) in the primary prevention setting. The purpose of this study was to examine the effect of ICDs, age, and multiple co-morbidities on survival in elderly patients who otherwise meet implantation criteria for primary prevention of sudden cardiac death. Patients >=80 years of age who received an ICD for LVEF <=35% at our institution from 2001 through 2008 (n = 99) were compared to a cohort of patients >=80 years of age with similarly low LVEF who did not receive an ICD (n = 53). Co-morbid conditions were assessed using the Charlson co morbidity index (CCI). The overall cohort (n = 152, 84 +/- 4 years old, 72% men, 87% with ischemic cardiomyopathy, LVEF 25 +/- 7%, CCI 5.9 +/- 3.2) was followed for 2.3 +/- 2.0 years. Patients with ICDs were younger (82 +/- 3 vs 86 +/- 4 years, p <0.001) and had fewer co-morbidities reflected in a lower CCI (5.3 +/- 3.1 vs 6.7 +/- 3.1, p = 0.021). Patients with ICDs also had a trend toward lower LVEF (24 +/- 6% vs 27 +/- 7%, p = 0.06). During follow-up 93 (61%) patients died, 58 (59%) in the ICD group and 35 (66%) in the no-ICD group. ICD recipients had better 1-year survival compared to patients with no ICD (72% vs 52%, p = 0.014). However, after adjusting for age, LVEF, glomerular filtration rate (GFR), and CCI using multivariate Cox models, an ICD did not confer any survival benefit (hazard ratio 0.71, 95% confidence interval 0.42 to 1.20, p = 0.20), whereas age (p = 0.043) and GFR (p = 0.006) were the only independent predictors of survival. In conclusion, age and GFR are the main determinant of survival in octogenarians and nonagenarians with LV dysfunction. After correcting for these parameters, an ICD does not appear to confer a survival benefit. PMID- 21640322 TI - Genetic variants at 13q12.12 are associated with high myopia in the Han Chinese population. AB - High myopia, which is extremely prevalent in the Chinese population, is one of the leading causes of blindness in the world. Genetic factors play a critical role in the development of the condition. To identify the genetic variants associated with high myopia in the Han Chinese, we conducted a genome-wide association study (GWAS) of 493,947 SNPs in 1088 individuals (419 cases and 669 controls) from a Han Chinese cohort and followed up on signals that were associated with p < 1.0 * 10(-4) in three independent cohorts (combined, 2803 cases and 5642 controls). We identified a significant association between high myopia and a variant at 13q12.12 (rs9318086, combined p = 1.91 * 10(-16), heterozygous odds ratio = 1.32, and homozygous odds ratio = 1.64). Furthermore, five additional SNPs (rs9510902, rs3794338, rs1886970, rs7325450, and rs7331047) in the same linkage disequilibrium (LD) block with rs9318086 also proved to be significantly associated with high myopia in the Han Chinese population; p values ranged from 5.46 * 10(-11) to 6.16 * 10(-16). This associated locus contains three genes-MIPEP, C1QTNF9B-AS1, and C1QTNF9B. MIPEP and C1QTNF9B were found to be expressed in the retina and retinal pigment epithelium (RPE) and are more likely than C1QTNF9B-AS1 to be associated with high myopia given the evidence of retinal signaling that controls eye growth. Our results suggest that the variants at 13q12.12 are associated with high myopia. PMID- 21640323 TI - Acute cerebrovascular manifestation of Takayasu arteritis. PMID- 21640324 TI - Hyperinsulinemia in nonalcoholic fatty liver disease in nondiabetic adults. PMID- 21640325 TI - Women were included twice in a meta-analysis. PMID- 21640326 TI - Malnutrition and hyponatremia in dialysis patients. PMID- 21640327 TI - Dabigatran falsely elevates point of care international normalized ratio results. PMID- 21640328 TI - The ten most annoying things that happen during my work day--and perhaps in yours as well. PMID- 21640329 TI - Career development of physician scientists: a survey of leaders in academic medicine. PMID- 21640330 TI - The role of breast cancer stem cells in metastasis and therapeutic implications. AB - Cancer stem cells (CSCs) possess the capacity to self-renew and to generate heterogeneous lineages of cancer cells that comprise tumors. A substantial body of evidence supports a model in which CSCs play a major role in the initiation, maintenance, and clinical outcome of cancers. In contrast, analysis of the role of CSCs in metastasis has been mainly conceptual and speculative. This review summarizes recent data that support the theory of CSCs as the source of metastatic lesions in breast cancer, with a focus on the key role of the microenvironment in the stemness-metastasis link. PMID- 21640331 TI - AP-1 transcription factor JunD confers protection from accelerated nephrotoxic nephritis and control podocyte-specific Vegfa expression. AB - Genetic investigation of crescentic glomerulonephritis (Crgn) susceptibility in the Wistar Kyoto rat, a strain uniquely susceptible to nephrotoxic nephritis (NTN), allowed us to positionally clone the activator protein-1 transcription factor Jund as a susceptibility gene associated with Crgn. To study the influence of Jund deficiency (Jund(-/-)) on immune-mediated renal disease, susceptibility to accelerated NTN was examined in Jund(-/-) mice and C57BL/6 wild-type (WT) controls. Jund(-/-) mice showed exacerbated glomerular crescent formation and macrophage infiltration, 10 days after NTN induction. Serum urea levels were also significantly increased in the Jund(-/-) mice compared with the WT controls. There was no evidence of immune response differences between Jund(-/-) and WT animals because the quantitative immunofluorescence for sheep and mouse IgG deposition in glomeruli was similar. Because murine Jund was inactivated by replacement with a bacterial LacZ reporter gene, we then investigated its glomerular expression by IHC and found that the Jund promoter is mainly active in Jund(-/-) podocytes. Furthermore, cultured glomeruli from Jund(-/-) mice showed relatively increased expression of vascular endothelial growth factor A (Vegfa), Cxcr4, and Cxcl12, well-known HIF target genes. Accordingly, small-interfering RNA-mediated JUND knockdown in conditionally immortalized human podocyte cell lines led to increased VEGFA and HIF1A expression. Our findings suggest that deficiency of Jund may cause increased oxidative stress in podocytes, leading to altered VEGFA expression and subsequent glomerular injury in Crgn. PMID- 21640332 TI - Thyroid effects and anticancer treatment. AB - Thyroid consequences of cancer therapy are multiple, better known after radiotherapy than after chemotherapy and recently described with targeted therapies. Cervical or total body irradiation may result in thyroid insufficiency or cancer. The consequences of treatment with new antiangiogenic drugs are under evaluation; however their effect on thyroid function is already well established. Thyroid dysfunction usually occurs late, several months or years after treatment and have to be depicted. There is an improvement in the overall survival of patients suffering from cancer and endocrinologists must be aware of the endocrine effects of treatments to propose an adequate survey and an appropriate treatment to improve well-being of patients. PMID- 21640334 TI - Direct projections from the central amygdaloid nucleus to the mesencephalic trigeminal nucleus in rats. AB - The amygdala is activated by fear and plays an important role in the emotional response to life-threatening situations. When rats feel threatened, they respond by biting fiercely. Bite strength is regulated by the trigeminal motor nucleus and the mesencephalic trigeminal nucleus (Me5). The Me5 relays proprioceptive signals from the masticatory muscles and the periodontal ligaments to the trigeminal motor and premotor nuclei. The amygdala projects to the trigeminal motor nucleus and the premotor reticular formation. However, it is unknown whether the amygdala projects directly to the Me5. In the present study, neurons of the central amygdaloid nucleus (ACe) were labeled following injection of a retrograde tracer, Fast Blue, into the caudal Me5, and fibers and terminal buttons from the ACe to the Me5 were examined after injections of an anterograde neuronal tracer, biotinylated dextran amine into the ACe. Furthermore, wheat germ agglutinin-conjugated to horseradish peroxidase was injected into the ACe, and labeled fibers and terminal buttons in the Me5 were examined by electron microscopy. Labeled terminal buttons on Me5 somata were more abundant in the caudal than the rostral Me5. Electron microscopic observation revealed that a part of these terminal buttons formed axo-somatic synapses. These results indicate that the ACe sends direct projections to the Me5, and suggest that the amygdala regulates bite strength by modifying neuronal activity in the Me5. PMID- 21640333 TI - Enhanced cocaine-conditioned place preference and associated brain regional levels of BDNF, p-ERK1/2 and p-Ser845-GluA1 in food-restricted rats. AB - Previously, a learning-free measure was used to demonstrate that chronic food restriction (FR) increases the reward magnitude of a wide range of abused drugs. Moreover, a variety of striatal neuroadaptations were detected in FR subjects, some of which are known to be involved in synaptic plasticity but have been ruled out as modulators of acute drug reward magnitude. Little is known about effects of FR on drug-conditioned place preference (CPP) and brain regional mechanisms that may enhance CPP in FR subjects. The purpose of the present study was to compare the expression and persistence of a conditioned place preference (CPP) induced by a relatively low dose of cocaine (7.0mg/kg, i.p.) in ad libitum fed (AL) and FR rats and take several brain regional biochemical measures following the first CPP conditioning session to probe candidate mechanisms that may underlie the more robust CPP observed in FR subjects. Behaviorally, AL subjects displayed a CPP upon initial testing which extinguished rapidly over the course of subsequent test sessions while CPP in FR subjects persisted. Despite previous reports of elevated BDNF protein in forebrain regions of FR rats, the FR protocol used in the present study did not alter BDNF levels in dorsal hippocampus, nucleus accumbens or medial prefrontal cortex. On the other hand, FR rats, whether injected with cocaine or vehicle, displayed elevated p-ERK1/2 and p Ser845-GluA1 in dorsal hippocampus. FR rats also displayed elevated p-ERK1/2 in medial prefrontal cortex and elevated p-ERK1 in nucleus accumbens, with further increases produced by cocaine. The one effect observed exclusively in cocaine treated FR rats was increased p-Ser845-GluA1 in nucleus accumbens. These findings suggest a number of avenues for continuing investigation with potential translational significance. PMID- 21640335 TI - Inclusion complex of 2-chlorobenzophenone with cyclomaltoheptaose (beta cyclodextrin): temperature, solvent effects and molecular modeling. AB - A thermodynamic study of the inclusion process between 2-chlorobenzophenone (2ClBP) and cyclomaltoheptaose (beta-cyclodextrin, beta-CD) was performed using UV-vis spectroscopy, reversed-phase liquid chromatography (RP-HPLC), and molecular modeling (PM6). Spectrophotometric measurements in aqueous solutions were performed at different temperatures. The stoichiometry of the complex is 1:1 and its apparent formation constant (K(c)) is 3846M(-1) at 30 degrees C. Temperature dependence of K(c) values revealed that both enthalpy (DeltaH degrees =-10.58kJ/mol) and entropy changes (DeltaS degrees =33.76J/Kmol) are favorable for the inclusion process in an aqueous medium. Encapsulation was also investigated using RP-HPLC (C18 column) with different mobile-phase compositions, to which beta-CD was added. The apparent formation constants in MeOH-H(2)O (K(F)) were dependent of the proportion of the mobile phase employed (50:50, 55:45, 60:40 and 65:35, v/v). The K(F) values were 419M(-1) (50% MeOH) and 166M(-1) (65% MeOH) at 30 degrees C. The thermodynamic parameters of the complex in an aqueous MeOH medium indicated that this process is largely driven by enthalpy change (DeltaH degrees =-27.25kJ/mol and DeltaS degrees =-45.12J/Kmol). The results of the study carried out with the PM6 semiempirical method showed that the energetically most favorable structure for the formation of the complex is the 'head up' orientation. PMID- 21640336 TI - Flavonoid C- and O-glycosides from the Mongolian medicinal plant Dianthus versicolor Fisch. AB - Eighteen flavonoids were identified from an aqueous extract of the aerial parts of Dianthus versicolor, a plant used in traditional Mongolian medicine against liver diseases. The flavonoid C- and O-glycosides isoorientin-7-O-rutinoside, isoorientin-7-O-rhamnosyl-galactoside, isovitexin-7-O-rutinoside, isovitexin-7-O rhamnosyl-galactoside, isoscoparin-7-O-rutinoside, isoscoparin-7-O-rhamnosyl galactoside, isoscoparin-7-O-galactoside, and isoorientin-7-O-galactoside were isolated and structurally elucidated. Their structures were established on the basis of extensive spectroscopic techniques including LC-UV-DAD, LC-MS(n), LC HRMS, 1D and 2D NMR spectroscopy, and by GC-MS analysis after hydrolysis. Flavonoids with such a high glycosylation pattern are rare within the genus Dianthus. Furthermore, isovitexin-7-O-glucoside (saponarin), isovitexin-2"-O rhamnoside, apigenin-6-glucoside (isovitexin), luteolin-7-O-glucoside, apigenin-7 O-glucoside, as well as the aglycons luteolin, apigenin, chrysoeriol, diosmetin, and acacetin were identified by TLC and LC-DAD-MS(n) in comparison to reference substances or literature data. The NMR data of seven structures have not been reported in the literature to date. PMID- 21640337 TI - Reducing cognitive biases in probabilistic reasoning by the use of logarithm formats. AB - Research on probability judgment has traditionally emphasized that people are susceptible to biases because they rely on "variable substitution": the assessment of normative variables is replaced by assessment of heuristic, subjective variables. A recent proposal is that many of these biases may rather derive from constraints on cognitive integration, where the capacity-limited and sequential nature of controlled judgment promotes linear additive integration, in contrast to many integration rules of probability theory (Juslin, Nilsson, & Winman, 2009). A key implication by this theory is that it should be possible to improve peoples' probabilistic reasoning by changing probability problems into logarithm formats that require additive rather than multiplicative integration. Three experiments demonstrate that recasting tasks in a way that allows people to arrive at the answers by additive integration decreases cognitive biases, and while people can rapidly learn to produce the correct answers in an additive formats, they have great difficulty doing so with a multiplicative format. PMID- 21640338 TI - Squeezing, striking, and vocalizing: Is number representation fundamentally spatial? AB - Numbers are fundamental entities in mathematics, but their cognitive bases are unclear. Abundant research points to linear space as a natural grounding for number representation. But, is number representation fundamentally spatial? We disentangle number representation from standard number-to-line reporting methods, and compare numerical estimations in educated participants using line-reporting with three nonspatial reporting conditions (squeezing, bell-striking, and vocalizing). All three cases of nonspatial-reporting consistently reproduced well established results obtained with number-line methods. Furthermore, unlike line reporting-and congruent with the psychophysical Weber-Fechner law-nonspatial reporting systematically produced logarithmic mappings for all nonsymbolic stimuli. Strikingly, linear mappings were obtained exclusively in conditions with culturally mediated elements (e.g., words). These results suggest that number representation is not fundamentally spatial, but builds on a deeper magnitude sense that manifests spatially and nonspatially mediated by magnitude, stimulus modality, and reporting condition. Number-to-space mappings-although ubiquitous in the modern world-do not seem to be rooted directly in brain evolution but have been culturally privileged and enhanced. PMID- 21640339 TI - Disordered eating attitudes in Egyptian antipsychotic naive patients with schizophrenia. AB - OBJECTIVES: The study aimed to test in a sample of Egyptian antipsychotic naive patients the hypotheses that the disordered eating attitudes co-occur with schizophrenia in a higher frequency than would be expected by chance in the general population and that the disordered eating comorbidity would be associated with more severe schizophrenia psychopathology. Previous studies have been mostly concerned with the impact of the antipsychotics. Studies relating abnormal eating behavior to the schizophrenia psychopathology rather than to its treatment are lacking. METHOD: In this case-control cross-sectional study, 50 consecutive antipsychotic naive patients, newly attending the psychiatric outpatient clinic, University Hospital, Zagazig, Egypt, with a Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, diagnosis of schizophrenia, were assessed by the Positive and Negative Syndrome Scale and compared with 50 nonpsychiatric controls using the Eating Attitudes Test (EAT40). RESULTS: Patients with schizophrenia had an EAT40 mean score (23.4 +/- 7.8) higher than that of controls (19.7 +/- 7.2) (P = .015). Prevalence of disordered eating (defined by a score of >= 30 on the EAT40) in these patients was higher than in the control group (30% vs 12%, P = .027). Comparison between schizophrenia patients with and without disordered eating showed no significant differences in demographic and a number of clinical variables, but they differed in their scores on lifestyle characteristics and anthropometric measures. The group of patients with disordered eating had also higher scores on total and all scale factors but not on the negative symptom scale. CONCLUSIONS: Data of this study show, perhaps for the first time, that "disordered" eating attitudes, as measured by the EAT40, are higher in a group of Egyptian patients with schizophrenia than in controls. However, the lack of difference between patients with and without disordered eating in terms of demographic and a number of clinical characteristics fail to explain the hypothesis that schizophrenia with disordered eating is a distinct subtype of schizophrenia. Data indicate, on the other hand, that the presence of disordered eating behavior in patients with schizophrenia is associated with the expression of more active psychotic symptoms. PMID- 21640340 TI - Childhood trauma, impulsivity, and executive functioning in crack cocaine users. AB - BACKGROUND: The use of crack cocaine is a major public health concern in Brazil and internationally. Recent data suggest that childhood trauma is associated with worse outcomes among cocaine users. This study had the objective of evaluating the relationship of childhood trauma with executive functioning and impulsivity in outpatients with crack cocaine use disorders. METHODS: This is a cross sectional study of 84 consecutive outpatients with a primary crack cocaine use disorder who sought treatment in Porto Alegre, Brazil. Childhood trauma was evaluated with the Childhood Trauma Questionnaire; executive functioning, with the Wisconsin Card Sorting Test; and impulsivity, with the Barratt Impulsivity Scale. RESULTS: Childhood trauma was strongly associated with executive dysfunction and impulsivity, even when controlled for possible confounders. CONCLUSIONS: Childhood trauma may be associated with executive dysfunction and impulsivity in crack cocaine users. The full impact of trauma needs to be further investigated in longitudinal studies. PMID- 21640341 TI - [Analysis of the drug interactions associated to domiciliary drug therapy in elderly hospitalized patients]. AB - OBJECTIVE: To determine the prevalence of potentially relevant drug-drug interactions associated with chronic treatment of elderly patients over 64-years of age on hospital admission and the factors associated with an increased presence of these. SUBJECTS AND METHODS: Cross-sectional observational study in a hospital referral area. All patients aged 65 or over admitted to the hospital in the last three months in 2009 were included. Based on the drug database of the General Council of Colleges of Pharmacy (BOT), drug-drug interactions and their potential clinical relevance were identified. To identify the variables associated with a higher prevalence of drug-drug interactions, analyses of correlation and of univariable linear regression and uni-and multivariable logistic regression analyses were performed using the SPSS, version 15.0. RESULTS: We analyzed the drug prescription data of 382 patients, whose mean age was 7.7 years. A total of 45.3% of patients had comorbidities and 78.8% had taken 5 or more drugs. We identified 272 clinically relevant drug-drug interactions that involved 159 patients (41.6%). Seven pharmacological groups accounted for 80.6% of the drug-drug interactions. The variables that had a statistically significant association to a higher prevalence of relevant interactions were polypharmacy, respiratory insufficiency, and treatment with proton-pump inhibitors, vitamin K antagonists, diuretics or anti-platelet drugs. CONCLUSIONS: A high prevalence of relevant drug-drug interactions was found in elderly hospitalized patients. Our findings suggest that prevention strategies should be implemented to avoid their associated adverse events, especially in high risk populations. PMID- 21640342 TI - Ectopic ovary confirmed by ovarian stimulation in a case of unicornuate uterus. AB - OBJECTIVE: To present a case of unicornuate uterus with ipsilateral ectopic ovary and renal agenesis. DESIGN: Case report. SETTING: Research hospital. PATIENT(S): A 26-year-old woman with a 5-year history of primary infertility was admitted to our clinic. A unicornuate uterus without contralateral horn, with ipsilateral ectopic ovary and renal agenesis, was diagnosed. INTERVENTION(S): Diagnostic laparoscopy, ovarian stimulation. MAIN OUTCOME MEASURE(S): During laparoscopy, a unicornuate uterus without a contralateral horn, together with ectopic ovary, was observed. RESULT(S): We visualized the right ectopic ovary on ultrasound after ovarian stimulation. CONCLUSION(S): We present a rare clinical entity that exhibits a unicornuate uterus without a contralateral horn, with ipsilateral right ectopic ovary and renal agenesis together. We conclude that ovarian stimulation is a useful approach in the diagnosis of ectopic ovaries. PMID- 21640343 TI - Endometrial Indian hedgehog expression is decreased in women with endometriosis. AB - Nuclear and cytoplasmic endometrial expression of Indian hedgehog increased from the late proliferative to mid and late secretory phases in 26 healthy volunteers compared with 30 women with endometriosis. The abnormal expression of Indian hedgehog protein in women with endometriosis suggests a resistance to P action. PMID- 21640344 TI - Activin-A is induced by interleukin-1beta and tumor necrosis factor-alpha and enhances the mRNA expression of interleukin-6 and protease-activated receptor-2 and proliferation of stromal cells from endometrioma. AB - OBJECTIVE: To examine the regulation and the function of activin-A in stromal cells derived from endometrioma. DESIGN: Molecular studies. SETTING: University research laboratory. PATIENT(S): Endometrioma stromal cells (EoSC) were obtained from 28 patients with ovarian endometrioma undergoing laparoscopy. INTERVENTION(S): EoSC were cultured with inflammatory stimuli or recombinant activin-A, followed by RNA extraction. MAIN OUTCOME MEASURE(S): Activin mRNA expression was evaluated by real-time reverse transcription-polymerase chain reaction (RT-PCR), and activin-A concentration of supernatant of cultured EoSC was evaluated by ELISA. Also, the effect of activin-A on EoSC was evaluated with real-time RT-PCR and cell proliferation assay. RESULT(S): Inflammatory stimuli, interleukin (IL) -1beta, and tumor necrosis factor (TNF) -alpha induced inhibin/activin-betaA subunit mRNA and activin-A protein expression in EoSC. Additionally, activin-A enhanced EoSC proliferation and increased the expression of IL-6 and protease-activated receptor (PAR)-2 mRNA. CONCLUSION(S): An in vitro study revealed that activin-A, which is induced by IL-1beta or TNF-alpha, might promote endometriosis by stimulating IL-6 and PAR-2 mRNA expression and increasing the proliferation of EoSC. PMID- 21640345 TI - A woman with premature ovarian failure induced by Tripterygium wilfordii Hook.f. gives birth to a healthy child. AB - OBJECTIVE: To inform clinicians about the reproductive damage of Tripterygium wilfordii Hook.f. (TW) associated with subsequent premature ovarian failure. DESIGN: Case report. SETTING: Reproductive medicine center in a university affiliated hospital. PATIENT(S): A 36-year-old infertile woman presenting amenorrhea and elevated FSH levels (65.56 mIU/mL) after using TW. INTERVENTION(S): Estradiol valerate, two month course of oral contraceptives, GnRH agonist, stimulation cycle with urine FSH, triggered ovulation using 5000 IU of hCG and IVF-ET. MAIN OUTCOME MEASURE(S): Serum hormone levels, antral follicle count (AFC), oocyte retrieval number, embryo quality and birth. RESULT(S): Serum hormone levels and AFC of the POF woman were restored, and she gave birth to a healthy child after IVF-ET. Through IVF-ET she was found to have had oocyte and granulosa cell damage because of TW. CONCLUSION(S): This is a real-life manifestation of reproductive damage of TW. The medicamentous amenorrhea, hormone levels and AFC decline induced by TW are reversible, but oocyte and granulosa cell damage may be irreversible. This will help clinicians to avoid using TW for nulligravida. PMID- 21640346 TI - Unusual appearances following intracapsular neck of femur fractures. AB - PURPOSE: The aim of the study was to investigate the possible outcomes of intracapsular femoral neck fractures managed non-operatively or with a simple form of internal fixation. METHODS: We reviewed 30 fractures and their post mortem reports from the Galler collection, a modern pathology reference series. RESULTS: Our investigation revealed unusual appearances of the hip following femoral neck fractures that are not usually seen in modern orthopaedic practice, including previously undescribed acetabularisation of the femur and ankyloses of ununited femoral heads. CONCLUSION: Appreciation of these appearances and an understanding of how these fractures may progress are important in the current diagnosis and management of delayed presentations, neglected or inadequately fixed subcapital fractures of the femoral neck. PMID- 21640347 TI - Long-form but not short-form Mini-Nutritional Assessment is appropriate for grading nutritional risk of patients on hemodialysis--a cross-sectional study. AB - BACKGROUND: Routine screening/assessment of protein-energy status is essential for preventing uremic malnutrition in patients on hemodialysis (HD). A simple, low cost, reliable and non-invasive tool is greatly desired. OBJECTIVE: This study aimed to evaluate the appropriateness of using the long-form (LF) and the short-form (SF) Mini Nutritional Assessment (MNA) for grading the risk of protein energy malnutrition in patients on HD. DESIGN AND SAMPLING: A cross-sectional study with purposive sampling. SETTING: A hospital-managed hemodialysis center. PARTICIPANTS: 152 adult ambulatory patients on hemodialysis. METHODS: The nutritional status of each patient was graded with MNA-LF and MNA-SF, each in two versions--a normalized-original (content-equivalent) version (by adopting population-specific anthropometric cut-off points) and an alternative version that replaced calf circumference for BMI in the scale. The SGA, serum albumin and serum creatinine served as references. Cross-tabulation test was used to evaluate the consistency of the versions. RESULTS: MNA-SF versions rated fewer HD subjects malnourished or at risk of malnutrition (32.2% and 24.3% for T1 and T2, respectively) compared to MNA-LF versions (40.8% and 36.2%) or the SGA (47.4%). MNA-SF versions (kappa=0.450 and 0.446) also did not perform as well as MNA-LF versions (kappa=0.734 and 0.666) in predicting the risk of malnutrition in HD patients using the SGA as the reference. MNA-SF also did not perform as well as the MNA-LF using serum albumin or serum creatinine as the reference. CONCLUSIONS: The MNA-LF is appropriate for predicting protein-energy malnutrition in HD patients but MNA-SF may under-rate these patients. Effort should be made to improve the MNA-SF for HD patients since the short-form is more time-efficient and thus, greatly desired in clinical practice. PMID- 21640348 TI - Effect of a traditional Mediterranean diet on apolipoproteins B, A-I, and their ratio: a randomized, controlled trial. AB - OBJECTIVES: Apolipoprotein (Apo)B, ApoA-I, and their ratio could predict coronary heart disease (CHD) risk more accurately than conventional lipid measurements. Our aim was to assess the effect of a traditional Mediterranean diet (TMD) on apolipoproteins. METHODS: High-cardiovascular risk subjects (n=551, 308 women and 243 men), aged 55-80 years, were recruited into a large, multicenter, randomized, controlled, parallel-group, clinical trial (The PREDIMED Study) aimed at testing the efficacy of TMD on primary cardiovascular disease prevention. Participants assigned to a low-fat diet (control) (n=177), or TMDs (TMD+virgin olive oil (VOO), n=181 or TMD+nuts, n=193) received nutritional education and either free VOO (ad libitum) or nuts (dose: 30 g/day). A 3-month evaluation was performed. RESULTS: Both TMDs promoted beneficial changes on classical cardiovascular risk factors. ApoA-I increased, and ApoB and ApoB/ApoA-I ratio decreased after TMD+VOO, the changes promoting a lower cardiometabolic risk. Changes in TMD+VOO versus low-fat diet were -2.9 mg/dL (95% CI, -5.6 to -0.08), 3.3mg/dL (95% CI, 0.84 to 5.8), and -0.03 mg/dL (-0.05 to -0.01) for ApoB, ApoA-I, and ApoB/ApoA-I ratio, respectively. CONCLUSIONS: Individuals at high-cardiovascular risk who improved their diet toward a TMD pattern rich in virgin olive oil, reduced their Apo B and ApoB/ApoA-I ratio and improved ApoA-I concentrations. PMID- 21640349 TI - Effect of alpha-glucosidase inhibitors on the progression of carotid intima-media thickness: a meta-analysis of randomized controlled trials. AB - BACKGROUND: It remains unclear whether lowering postprandial glucose by alpha glucosidase inhibitors (alpha-GIs) can prevent the progression of carotid intima media thickness (IMT). METHODS AND RESULTS: We performed a meta-analysis of all relevant randomized controlled trials (RCTs) to evaluate the effect of alpha-GIs on the progression of carotid IMT. Five RCTs with 411 patients were included in this study. Alpha-GIs therapy was associated with a significant reduction in the annual progression of carotid IMT (WMD, -0.06 mm/year, 95% CI -0.11, -0.01; P=0.02) and the progression in carotid IMT at the end of follow-up (WMD, -0.07 mm, 95% CI -0.12, -0.02; P=0.003). In subgroup analysis, alpha-GIs therapy was associated with a significant reduction in the annual progression of carotid IMT in patients with type 2 diabetes mellitus (T2DM) (WMD, -0.08 mm/year, 95% CI 0.10, -0.06; P<0.00001), and in the progression of carotid IMT in patients with impaired glucose tolerance (IGT) at the end of follow-up (WMD, -0.03 mm, 95% CI 0.05, -0.01; P=0.01). Alpha-GIs treatment was associated with significant increase in HDL-C (WMD 1.56 mg/dl, 95% CI 0.09, 3.03; P=0.04) and decrease in basal immunoreactive insulin as well as had favorable trends towards reducing HbA1c, triglyceride and diastolic blood pressure. CONCLUSION: It suggests that alpha-GIs therapy may be an effective strategy in preventing the progression of carotid IMT in patients with IGT or T2DM. It partially contributes to the improvement in atherogenic metabolic parameters induced by alpha-GIs. More studies, especially large multi-centre RCTs, are still warranted to further clarify the anti-atherosclerotic effect of alpha-GIs. PMID- 21640350 TI - Relationship between plasma coenzyme Q10, asymmetric dimethylarginine and arterial stiffness in patients with phenotypic or genotypic familial hypercholesterolemia on long-term statin therapy. AB - OBJECTIVE: We investigated whether statin-treated heterozygous familial hypercholesterolemic (FH) patients have lower plasma coenzyme Q(10) (CoQ(10)) levels than low-density lipoprotein receptor (LDLR) mutation negative FH patients on equivalent statin doses, and whether lower CoQ(10) concentrations are associated with increased arterial stiffness. METHODS: Thirty LDLR mutation negative patients with clinical FH and 30 mutation positive FH patients matched for gender, statin duration and dose, and a further 30 controls were studied. Plasma CoQ(10) and asymmetric dimethylarginine (ADMA) levels were measured by HPLC and the augmentation index by pulse wave analysis. RESULTS: Plasma CoQ(10) levels, and the ratios of CoQ(10) to total cholesterol and LDL-cholesterol were similar in treated FH patients with identified LDLR mutations to mutation negative patients on equivalent doses of statin therapy (p>0.05). CoQ(10) and lipid levels were also comparable to controls not using any lipid modifying treatment. Arterial stiffness was higher in mutation negative patients (p=0.04) and there was a trend for an increase in mutation positive patients (p=0.09). ADMA was higher in the mutation positive group (p<0.01). The augmentation index corrected for age, blood pressure, and heart rate, was negatively correlated with plasma CoQ(10) within FH patients (p<0.05). CONCLUSION: Long-term, high-dose statin therapy does not lead to subnormal CoQ(10) concentrations in patients with phenotypic or genotypic FH. Arterial stiffness is elevated in FH patients compared to untreated controls, and low CoQ(10) levels are associated with increased arterial stiffness. CoQ(10) supplementation trials are warranted in FH patients. PMID- 21640351 TI - Periodontitis and blood pressure: the concept of dental hypertension. AB - Chronic periodontitis is a common inflammatory disorder that is being contemplated as a risk factor for atherosclerotic complications. Current epidemiological evidence also supports its potential association with increases in blood pressure levels and hypertension prevalence. Furthermore, data from cross-sectional studies suggest that in hypertensive subjects periodontitis may enhance the risk and degree of target organ damage. A possible pathogenetic background of an effect of periodontitis on blood pressure should include the systemic generalization of the local oral inflammation, the role of the host immune response, the direct microbial effect on the vascular system and alterations in endothelial function. Inversely, the concept of hypertension unfavorably affecting periodontal tissues cannot be excluded. The two conditions share multiple common risk factors that should be readily controlled for when assessing a possible association. Thoroughly designed prospective and interventional trials are needed in order to determine the impact of periodontitis on blood pressure regulation and incident hypertension and its integration in the clinical approach of both dental and hypertensive patients. PMID- 21640352 TI - An algorithm to correct for camera vibrations in optical motion tracking systems. AB - Recording and reconstruction of 3D motion capturing data relies on fixed, static camera positions with given inter-camera distances in a laboratory frame. To overcome this limitation, we present a correction algorithm that allows us to address camera movements in moving camera setups. Camera vibrations are identified by comparison of specialized target positions in dynamic measurements with their respective positions in static trials. This results in a 2D shift vector Deltaw with which the individual camera streams are corrected. The capabilities of this vibration reduction procedure are demonstrated in a test setup of four cameras that are (i) separately and (ii) simultaneously perturbed while capturing a static test object. In the former case, the correction algorithm is capable of reducing the reconstruction residuals to the order of the calibrations residual and enables reconstruction in the latter case, which is impossible without any correction. This approach extends the application of marker-based infrared motion tracking to moving and even accelerated camera setups. PMID- 21640353 TI - Shoulder muscle function depends on elbow joint position: an illustration of dynamic coupling in the upper limb. AB - Shoulder muscle function has been documented based on muscle moment arms, lines of action and muscle contributions to contact force at the glenohumeral joint. At present, however, the contributions of individual muscles to shoulder joint motion have not been investigated, and the effects of shoulder and elbow joint position on shoulder muscle function are not well understood. The aims of this study were to compute the contributions of individual muscles to motion of the glenohumeral joint during abduction, and to examine the effect of elbow flexion on shoulder muscle function. A three-dimensional musculoskeletal model of the upper limb was used to determine the contributions of 18 major muscles and muscle sub-regions of the shoulder to glenohumeral joint motion during abduction. Muscle function was found to depend strongly on both shoulder and elbow joint positions. When the elbow was extended, the middle and anterior deltoid and supraspinatus were the greatest contributors to angular acceleration of the shoulder in abduction. In contrast, when the elbow was flexed at 90 degrees , the anterior deltoid and subscapularis were the greatest contributors to joint angular acceleration in abduction. This dependence of shoulder muscle function on elbow joint position is explained by the existence of dynamic coupling in multi-joint musculoskeletal systems. The extent to which dynamic coupling affects shoulder muscle function, and therefore movement control, is determined by the structure of the inverse mass matrix, which depends on the configuration of the joints. The data provided may assist in the diagnosis of abnormal shoulder function, for example, due to muscle paralysis or in the case of full-thickness rotator cuff tears. PMID- 21640354 TI - A method to characterize in vivo tendon force-strain relationship by combining ultrasonography, motion capture and loading rates. AB - The ultrasonography contributes to investigate in vivo tendon force-strain relationship during isometric contraction. In previous studies, different methods are available to estimate the tendon strain, using different loading rates and models to fit the tendon force-strain relationship. This study was aimed to propose a standard method to characterize the in vivo tendon force-strain relationship. We investigated the influence on the force-strain relationship for medialis gastrocnemius (MG) of (1) one method which takes into account probe and joint movements to estimate the instantaneous tendon length, (2) models used to fit the force-strain relationship for uniaxial test (polynomial vs. Ogden), and (3) the loading rate on tendon strain. Subjects performed ramp-up contraction during isometric contractions at two different target speeds: 1.5s and minimal time with ultrasound probe fixed over the muscle-tendon junction of the MG muscle. The used method requires three markers on ultrasound probe and a marker on calcaneum to take into account all movements, and was compared to the strain estimated using ultrasound images only. The method using ultrasound image only overestimated the tendon strain from 40% of maximal force. The polynomial model showed similar fitting results than the Ogden model (R2=0.98). A loading rate effect was found on tendon strain, showing a higher strain when loading rate decreases. The characterization of tendon force-strain relationship needs to be standardized by taking into account all movements to estimate tendon strain and controlling the loading rate. The polynomial model appears to be appropriate to represent the tendon force-strain relationship. PMID- 21640355 TI - Screening, identification and quantification of glucosinolates in black radish (Raphanus sativus L. niger) based dietary supplements using liquid chromatography coupled with a photodiode array and liquid chromatography-mass spectrometry. AB - The glucosinolate profile of black radish (Raphanus sativus L. niger) based dietary supplements has been investigated by HPLC-PDA, LC-ESI-MS/MS and LC-APCI MS/MS systems. Optimization of the MS/MS parameters and LC conditions was performed using sinigrin reference standard and rapeseed certified reference material (BC190) respectively. An LC-ESI-MS/MS system was used to detect (screen) and identify the naturally occurring intact glucosinolates (GLs). The intact GLs identified were then desulfated and quantified on an HPLC-PDA system as desulfo glucosinolates (DS-GLs). Prior to quantification, the DS-GLs were identified using an APCI-MS/MS. The HPLC-PDA method performance criteria were evaluated using glucotropaeolin potassium salt. The validated method was applied for the analysis of six dietary supplements. In total, six glucosinolates were identified and quantified in the dietary supplements; glucoraphasatin (0.2-0.48 mg/g), glucosisaustricin (0.37-0.91 mg/g), glucoraphenin (0.84-1.27 mg/g), glucoputrajivin (0.14-0.28 mg/g), glucosisymbrin (0.70-0.99 mg/g) and gluconasturtiin (0.06-0.12 mg/g). Glucoraphenin was the most abundant glucosinolate in all samples. PMID- 21640356 TI - Poly(lactic acid)-coated mesoporous silica nanosphere for controlled release of venlafaxine. AB - Two types of mesoporous silica nanospheres (MSNs) were synthesized for use as controlled-release agents. One was prepared by grafting with 5,6 dihydroxyhexylsilane (DH-MSN) and the other one by further coating with cholic acid-crosslinked poly(lactic acid) (CA-PLA-MSN). We studied the release of the antidepressant venlafaxine from each of the materials in simulated gastric fluid (SGF), in simulated gastric acid solution (SGA), and in simulated intestinal fluid without pancreatin (SIF). The CA-PLA-MSN material was able to significantly delay the release of the drug in intestinal condition compared with gastric acid surrounding due to the fast decomposition rate of PLA in gastric acid. Moreover, it successfully avoided the initial burst to a certain extent in SGF. The enzyme pepsin played a favorable obstruct role in both DH-MSN and CA-PLA-MSN systems to reduce release rate. A model based on Weibull model was built to fit the release results, and based on it, the mechanisms about release processes were brought out tentatively. PMID- 21640357 TI - Poly(zinc dimethacrylate) as a precursor in the low-temperature formation of ZnO nanoparticles. AB - We present a simple, low-temperature synthesis of pure ZnO nanoparticles and polymer-ZnO hybrid materials formed by the NaOH-mediated conversion of poly(zinc dimethacrylate) in 1-butanol. The polymer poly(zinc dimethacrylate) was used as a precursor to prepare neat ZnO particles. It has a double role in the ZnO formation process, acting as a template and simultaneously controlling the crystal growth. The obtained single-crystalline ZnO nanorods show a low tendency to aggregate. The reaction mechanism of ZnO formation was proposed on the basis of a model system of the base-mediated conversion of a monomer zinc dimethacrylate Zn(MA)(2). PMID- 21640358 TI - Mechanisms of TiO2 nanoparticle transport in porous media: role of solution chemistry, nanoparticle concentration, and flowrate. AB - The role of solution chemistry, nanoparticle concentration and hydrodynamic effects in the transport and deposition of TiO(2) nanoparticles through porous media has been systematically investigated. Two solution chemistry variables, pH and ionic strength (IS), showed a significant influence on the transport due to their involvement in the aggregation of the nanoparticles and interaction with quartz sand. An electrostatically unfavorable condition for deposition existed at pH 7, at which the greatest retention occurred in the column, likely due to aggregation (>1000 nm) and straining effects. Under electrostatically favorable conditions (pH 5) significant elution from the column was observed and attributed to smaller aggregate size (~300 nm) and blocking effects. Nanoparticle concentration was found to contribute to the increased breakthrough of nanoparticles at pH 5 due to blocking and subsequent particle-particle repulsion. Increased flowrate resulted in greater elution of nanoparticles due to hydrodynamic forces acting on aggregates and subsequently contributed to blocking. Overall, a combination of mechanisms including straining, blocking, and DLVO-type forces were involved over the range of solution chemistry and nanoparticle concentrations tested. Consideration of these mechanisms is necessary for improved removal of TiO(2) nanoparticles via filtration and reliable prediction of transport of these potentially problematic nanoparticles through the subsurface. PMID- 21640359 TI - Systemic blood pressure profile correlates with cardiac 123I-MIBG uptake in patients with Parkinson's disease. AB - To examine the correlation between the systemic blood pressure profile and cardiac (123)I-metaiodobenzylguanidine (MIBG) uptake in patients with Parkinson's disease (PD), we monitored circadian blood pressure patterns of 37 PD patients of 49 to 85 years of age (mean, 71.8+/-8.4 years) using a portable blood pressure monitoring device. The duration of PD was 0.5 to 15 years, and the disability level (modified Hoehn and Yahr stage) ranged from 1.0 to 4.0 (mean, 2.7+/-0.7). There were 37 age- and sex-matched control subjects. Cardiac MIBG scintigraphy was performed for the 37 PD patients. Based on the nocturnal fall in mean arterial blood pressure (MABP), we classified patients into extreme dippers (nocturnal reduction of MABP >20%), dippers (>10% but <20%), nondippers (<10% but >0%), and inverted dippers (<0%). Average 24-hour MABP values revealed reduced BP variability in PD patients. The percentage nocturnal fall in MABP was significantly different between PD patients and control subjects (p<0.05). Significant correlations were found between % MABP reduction and the heart-to mediastinum (H/M) ratio on early and delayed images (p<0.01). The UPDR motor score, early and delay H/M ratios were also significantly different between patients who were and were not dippers (p<0.05). The present results reported for the first time a significant correlation between the systemic blood pressure profile and cardiac (123)I-MIBG uptake in patients with PD. The degeneration between the brainstem and the postganglionic neurons of myocardial sympathetic nerves may progress in parallel in patients with PD. PMID- 21640360 TI - Metabolic and endocrine effects of long-chain versus essential omega-3 polyunsaturated fatty acids in polycystic ovary syndrome. AB - The objective of the study was to compare the effects of essential vs long-chain omega (n)-3 polyunsaturated fatty acids (PUFAs) in polycystic ovary syndrome. In this 6-week, prospective, double-blinded, placebo (soybean oil)-controlled study, 51 completers received 3.5 g n-3 PUFA per day (essential PUFA from flaxseed oil or long-chain PUFA from fish oil). Anthropometric variables, cardiovascular risk factors, and androgens were measured; oral glucose tolerance test (OGTT) and frequently sampled intravenous GTT (IVGTT) were conducted at baseline and 6 weeks. Between-group comparisons showed significant differences in serum triglyceride response (P = .0368), whereas the changes in disposition index also tended to differ (P = .0621). When within-group changes (after vs before intervention) were considered, fish oil and flaxseed oil lowered serum triglyceride (P = .0154 and P = .0176, respectively). Fish oil increased glucose at 120 minutes of OGTT (P = .0355), decreased the Matsuda index (P = .0378), and tended to decrease acute insulin response during IVGTT (P = .0871). Soybean oil increased glucose at 30 (P = .0030) and 60 minutes (P = .0121) and AUC for glucose (P = .0122) during OGTT, tended to decrease acute insulin response during IVGTT (P = .0848), reduced testosterone (P = .0216), and tended to reduce sex hormone-binding globulin (P = .0858). Fasting glucose, insulin, adiponectin, leptin, or high-sensitivity C-reactive protein did not change with any intervention. Long-chain vs essential n-3 PUFA-rich oils have distinct metabolic and endocrine effects in polycystic ovary syndrome; and therefore, they should not be used interchangeably. PMID- 21640362 TI - [Hybrid treatment of a symptomatic aneurysm of a Kommerell's diverticulum]. AB - Left aortic arch with aberrant right subclavian artery is the most common congenital vascular anomaly involving the aortic arch. In 60% of cases, the aberrant right subclavian artery arises from a dilated segment of the aortic arch called Kommerell's diverticulum. Aneurysm of the aberrant right subclavian artery is rare; this condition could remain clinically silent or it could originate nonspecific symptoms. Rupture of an aneurysm of the aberrant right subclavian artery is associated with high mortality. Although there are no exact criteria to indicate the treatment of this condition, repair of the aneurysm is recommended when symptoms occur or when it reaches a size of 30 mm to 50mm. The malformation can be suspected at plain-film X-ray examination, but magnetic resonance imaging (MRI) or computed tomography (CT) are the imaging tests of choice for the diagnosis and for planning treatment. We present the case of a patient with an aberrant right subclavian artery with a small calcified aneurysm in a Kommerell's diverticulum that caused chest pain and dysphagia; the patient underwent a procedure combining endovascular and surgical treatment. PMID- 21640361 TI - Nebivolol improves insulin sensitivity in the TGR(Ren2)27 rat. AB - Hypertension is often associated with increased oxidative stress and systemic insulin resistance. Use of beta-adrenergic receptor blockers in hypertension is limited because of potential negative influence on insulin sensitivity and glucose homeostasis. We sought to determine the impact of nebivolol, a selective vasodilatory beta1-adrenergic blocker, on whole-body insulin sensitivity, skeletal muscle oxidative stress, insulin signaling, and glucose transport in the transgenic TG(mRen2)27 rat (Ren2). This rodent model manifests increased tissue renin angiotensin expression, excess oxidative stress, and whole-body insulin resistance. Young (age, 6-9 weeks) Ren2 and age-matched Sprague-Dawley control rats were treated with nebivolol 10 mg/(kg d) or placebo for 21 days. Basal measurements were obtained for glucose and insulin to calculate the homeostasis model assessment. In addition, insulin metabolic signaling, nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity, reactive oxygen species, and ultrastructural changes as evaluated by transmission electron microscopy were examined ex vivo in skeletal muscle tissue. The Ren2 rat demonstrated systemic insulin resistance as examined by the homeostasis model assessment, along with impaired insulin metabolic signaling in skeletal muscle. This was associated with increased oxidative stress and mitochondrial remodeling. Treatment with nebivolol was associated with improvement in insulin resistance and decreased NADPH oxidase activity/levels of reactive oxygen species in skeletal muscle tissue. Nebivolol treatment for 3 weeks reduces NADPH oxidase activity and improves systemic insulin resistance in concert with reduced oxidative stress in skeletal muscle in a young rodent model of hypertension, insulin resistance, and enhanced tissue RAS expression. PMID- 21640363 TI - [Magnetic resonance imaging and spectrometric study of the distribution of thermotherapeutic magnetofluid after intra-arterial administration in an experimental model of liver metastases]. AB - OBJECTIVE: To use imaging and laboratory techniques to evaluate the vascular distribution of magnetofluid in a rat model of liver metastases. MATERIAL AND METHODS: The livers of 33 WAG/Rij Crl rats were seeded with CC-531 colorectal cancer cells. After we checked tumor development, 10 rats received hepatic intra arterial infusions of Lipiodol((r)) with nanoparticles of Fe(3)O(4) in suspension, and 5 were reserved as controls. Axial STIR (TR: 3,600 ms/TE: 29 ms/TI: 130 ms) and gradient-echo (GE) (120/4 and 120/14) MRI sequences were acquired on a 1.5 T scanner. After necropsy, rats were classified into one of two stages according to tumor development: early (<10 metastases, each < 3mm) or advanced (>10 metastases, each >3 mm). Samples of liver and of metastases were taken from the 15 animals for quantification of iron concentrations by inductively coupled plasma mass spectrometry (ICP-MS). The data were analyzed using nonparametric tests; values of p < 0.05 were considered significant. RESULTS: Five animals had early tumor development and five had advanced tumor development. In the GE sequences, early stage metastases showed homogeneous signal reduction attributable to the presence of magnetofluid. Spectrometry found significant differences between the iron concentration in rats with early stage metastases and controls (p=0.002) as well as between rats with early stage metastases and those with late stage metastases (p=0.001). The ratio of exogenous iron in metastases and in liver in early stage rats was 2.6:1. The concentration of exogenous iron in the liver was significantly different from that in tumors only in early stage animals (p=0.043). CONCLUSIONS: MRI and spectrometry made it possible to evaluate the vascular distribution of magnetofluid in the liver and revealed the differences in its affinity for metastases in different stages of disease. PMID- 21640364 TI - [Hydroxyapatite bone cement for sinus floor augmentation with simultaneous implant placement. A preliminary study]. PMID- 21640365 TI - Antoine Ritti (1844-1920), forgotten alienist and innovative theorist on the pathophysiology of hallucinations. AB - Antoine Ritti (1844-1920), now forgotten, was an "alienist" or psychiatrist who formulated a theory of hallucination based on thalamic dysfunction, as described in his thesis defended in 1874. Ritti was a student of Jules Luys and used the anatomical-functional discoveries of his teacher to explain that an automatic activity in the thalamus, by stimulating the cortex without reception of sensory information, created autonomous representations, perceived by the patient but not by his entourage, a process occurring spontaneously to some degree. Hence, Ritti seems the first author to introduce the concept of sensory deprivation and release of subcortical function into the pathophysiology of hallucinations. This innovative theory, which gave subcortical structures a role in high-level cognitive function, is very resonant today but was ignored for several decades after Ritti published his work. PMID- 21640366 TI - Marc Dax and the discovery of the lateralisation of language in the left cerebral hemisphere. AB - In 1836, Marc Dax presented a revolutionary conclusion based on both his clinical observations and reports from the literature: the localisation of language in the left hemisphere. Twenty-seven years later, his son, Gustave Dax, extended and enriched his work. Their involvement in 19th century neuropsychology is impressive and recognition should have been considerable. However, according to the vast majority of historians of neurology, neuropsychology and aphasiology, the priority in establishing the dominance of the left brain hemisphere for speech goes to Paul Broca in an undivided way. Is it possible that Marc Dax's and his son, Gustave Dax's works were entirely unknown? Were they known and utterly forgotten? Were they ignored? How did we get to know that they existed? The aim of the present paper is to try to answer those and other questions and to demonstrate that there is abundance of data pointing out that the priority of the discovery of left hemisphere dominance for speech ought to be, at least, shared by Dax and Broca. PMID- 21640367 TI - Participation of the Cowpea mosaic virus protease in eliciting extreme resistance. AB - Extreme resistance of Arlington line cowpea (Vigna unguiculata) to Cowpea mosaic virus (CPMV) is under control of a dominant locus designated Cpa. We transiently expressed, using Tomato bushy stunt virus (TBSV) vectors and Agrobacterium tumefaciens, in nearly isogenic Cpa/Cpa and cpa/cpa cowpea lines, sequences from RNA1, the larger of two CPMV genomic RNAs. Activation of a Cpa-specific response mapped to the CPMV 24K protease (24KPro). Mutational analysis of the 24KPro gene implicated protease activity, rather than 24KPro structure, in Cpa-mediated recognition of CPMV invasion. A 24KPro with alanine replacing the active site cysteine [24KPro(C-A)], but not wildtype 24KPro, accumulated after agroinfiltration of the corresponding binary vector constructions into Cpa/Cpa cowpea. In cpa/cpa cowpea, both protease versions accumulated, with 24KPro(C-A) in greater abundance. Thus, enzymically active 24KPro was recognized by both cowpea genotypes, but in Cpa/Cpa cowpea the suppression of 24KPro accumulation was very strong, consistent with extreme resistance to CPMV. PMID- 21640368 TI - Distribution and potential significance of a gull fecal marker in urban coastal and riverine areas of southern Ontario, Canada. AB - To better understand the distribution of gull fecal contamination in urban areas of southern Ontario, we used gull-specific PCR and qPCR assays against 1309 water samples collected from 15 urban coastal and riverine locations during 2007. Approximately, 58% of the water samples tested positive for the gull-assay. Locations observed to have higher numbers of gulls and their fecal droppings had a higher frequency of occurrence of the gull marker and a higher gull marker qPCR signal than areas observed to be less impacted by gulls. Lower gull marker occurrence and lower qPCR signals were associated with municipal wastewater (7.4%) and urban stormwater effluents (29.5%). Overall, there were no statistically significant differences in gull marker occurrence at beach sites for pore water, ankle, and chest-depth samples, although signals were generally higher in interstitial beach sand pore water and ankle-depth water than in chest depth water samples. Overall, the results indicated that gull fecal pollution is widespread in urban coastal and riverine areas in southern Ontario and that it significantly contributes to fecal indicator bacterial loads. PMID- 21640369 TI - Root features related to plant growth and nutrient removal of 35 wetland plants. AB - Morphological, structural, and eco-physiological features of roots, nutrient removal, and correlation between the indices were comparatively studied for 35 emergent wetland plants in small-scale wetlands for further investigation into the hypothesis of two types of wetland plant roots (Chen et al., 2004). Significant differences in root morphological, structural, and eco-physiological features were found among the 35 species. They were divided into two types: fibrous-root plants and thick-root plants. The fibrous-root plants had most or all roots of diameter (D) <= 1 mm. Roots of D > 1 mm also had many fine and long lateral roots of D <= 1 mm. The roots of these plants were long and had a thin epidermis and a low degree of lignification. The roots of the thick-root plants were almost all thicker than 1 mm, and generally had no further fine lateral roots. The roots were short, smooth, and fleshy, and had a thick epidermis. Root porosity of the fibrous-root plants was higher than that of the thick-root plants (p = 0.001). The aerenchyma of the fibrous-root plants was composed of large cavities which were formed from many small cavities, and distributed radially between the exodermis and vascular tissues. The aerenchyma of the thick-root plants had a large number of small cavities which were distributed in the mediopellis. The fibrous-root plants had a significantly larger root biomass of D <= 1 mm, of 1 mm < D < 3 mm, above-ground biomass, total biomass, and longer root system, but shorter root longevity than those of the thick-root plants (p = 0.003, 0.018, 0.020, 0.032, 0.042, 0.001). The fibrous-root plants also had significantly higher radial oxygen loss (ROL), root activity, photosynthetic rate, transpiration rate, and removal rates of total nitrogen and total phosphorus than the thick-root plants (p = 0.001, 0.008, 0.010, 0.004, 0.020, 0.002). The results indicate that significantly different root morphological and structural features existed among different wetland plants, and these features had a close relationship to nutrient removal capacity. PMID- 21640370 TI - Oxidative stress, cytoxicity, and cell mortality induced by nano-sized lead in aqueous suspensions. AB - This paper reports on the effect of aqueous and nano-particulated Pb on oxidative stress (lipid peroxidation), cytoxicity, and cell mortality. As determined by the Thiobarbituric Acid Reactive Substances (TBARS) method, only 6h after incubation aqueous suspensions bearing nano-sized PbO(2), soluble Pb(II), and brain homogenate only suspensions, were determined to contain as much as ca. 7, 5, and 1 nmol TBARS mg protein(-1), respectively. Exposure of human cells (central nervous system, prostate, leukemia, colon, breast, lung cells) to nano-PbO(2) led to cell-growth inhibition values (%) ca. <=18.7%. Finally, as estimated by the Artemia salina test, cell mortality values were found to show high-survival larvae rates. Microscopic observations revealed that Pb particles were swallowed, but caused no mortality, however. PMID- 21640371 TI - Effects on aquatic and human health due to large scale bioenergy crop expansion. AB - In this study, the environmental impacts of large scale bioenergy crops were evaluated using the Soil and Water Assessment Tool (SWAT). Daily pesticide concentration data for a study area consisting of four large watersheds located in Michigan (totaling 53,358 km2) was estimated over a six year period (2000 2005). Model outputs for atrazine, bromoxynil, glyphosate, metolachlor, pendimethalin, sethoxydim, triflualin, and 2,4-D model output were used to predict the possible long-term implications that large-scale bioenergy crop expansion may have on the bluegill (Lepomis macrochirus) and humans. Threshold toxicity levels were obtained for the bluegill and for human consumption for all pesticides being evaluated through an extensive literature review. Model output was compared to each toxicity level for the suggested exposure time (96-hour for bluegill and 24-hour for humans). The results suggest that traditional intensive row crops such as canola, corn and sorghum may negatively impact aquatic life, and in most cases affect the safe drinking water availability. The continuous corn rotation, the most representative rotation for current agricultural practices for a starch-based ethanol economy, delivers the highest concentrations of glyphosate to the stream. In addition, continuous canola contributed to a concentration of 1.11 ppm of trifluralin, a highly toxic herbicide, which is 8.7 times the 96-hour ecotoxicity of bluegills and 21 times the safe drinking water level. Also during the period of study, continuous corn resulted in the impairment of 541,152 km of stream. However, there is promise with second generation lignocellulosic bioenergy crops such as switchgrass, which resulted in a 171,667 km reduction in total stream length that exceeds the human threshold criteria, as compared to the base scenario. Results of this study may be useful in determining the suitability of bioenergy crop rotations and aid in decision making regarding the adaptation of large-scale bioenergy cropping systems. PMID- 21640372 TI - Prognosis of ovarian clear cell carcinoma compared to other histological subtypes: a meta-analysis. AB - OBJECTIVE: To compare the survival outcome between clear cell carcinoma (CCC) and other histological subtypes in epithelial ovarian carcinoma (EOC). METHODS: From January 1974 to February 2011, we identified a total of 31,800 (CCC; 2152, non CCC; 29648) patients from 12 studies meeting the inclusion criteria. RESULTS: Heterogeneity tests demonstrated significant between-study variation (I(2)=92.1%) with no significant difference in hazard ratio (HR) for death between CCC and non CCC (HR; 1.16, 95% CI; 0.85-1.57, random-effects model). Comparing the HR based on stage I+II, and stage III+IV, between CCC and non-CCC, showed that CCC patients had a higher hazard rate for death than those with non-CCC of the ovary (stage I+II; HR; 1.17, 95% CI; 1.01-1.36, stage III+IV; HR; 1.65, 95% CI; 1.52 1.79). In a comparison of CCC and serous EOC, advanced stage (III and IV) CCC only showed a poorer hazard rate for death than serous EOC (HR; 1.71, 95% CI; 1.57-1.86). CONCLUSION: This analysis suggests that ovarian CCC patients had poorer prognosis than those with other histological subtypes of EOC, especially in advanced EOC stages. Different treatment strategies may be needed for patients with ovarian CCC. PMID- 21640373 TI - FOXL2 C402G mutation detection using MALDI-TOF-MS in DNA extracted from Israeli granulosa cell tumors. AB - OBJECTIVE: To develop a rapid, sensitive and reliable method to detect FOXL2 C402G mutation in granulosa cell tumor (GCT) and to investigate the prevalence of FOXL2 mutation in granulose cell tumors among Israeli patients. METHODS: We designed and optimized a matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) genotyping assay to detect FOXL2 C402G mutation in DNA isolated from formalin-fixed paraffin-embedded tissue samples. We examined 20 tumor samples obtained from Israeli patients diagnosed with granulose cell tumor. RESULTS: Eighteen out of 20 samples were found to harbor FOXL2 C402G mutation. Pathological review of the two tumors harboring wild type FOXL2 (C402) concluded that they were adenocarcinomas and has been misclassified at initial diagnosis. We found that the prevalence of FOXL2 mutations among Israeli patients with GCT (100%) is similar to previous reports. CONCLUSIONS: Our results indicate that the FOXL2 mutations can be reliably detected by MALDI-TOF-MS genotyping. MALDI-TOF-MS genotyping is a simple, robust and highly sensitive method to detect FOXL2 C402G mutation. Our results confirm previous studies reporting over 95% prevalence of FOXL2 mutation in GCT. Furthermore, we suggest that testing for the presence of the FOXL2 C402G mutation may improve diagnostic accuracy. PMID- 21640375 TI - Iodine status of UK schoolgirls: a cross-sectional survey. AB - BACKGROUND: Iodine deficiency is the most common cause of preventable mental impairment worldwide. It is defined by WHO as mild if the population median urinary iodine excretion is 50-99 MUg/L, moderate if 20-49 MUg/L, and severe if less than 20 MUg/L. No contemporary data are available for the UK, which has no programme of food or salt iodination. We aimed to assess the current iodine status of the UK population. METHODS: In this cross-sectional survey, we systematically assessed iodine status in schoolgirls aged 14-15 years attending secondary school in nine UK centres. Urinary iodine concentrations and tap water iodine concentrations were measured in June-July, 2009, and November-December, 2009. Ethnic origin, postcode, and a validated diet questionnaire assessing sources of iodine were recorded. FINDINGS: 810 participants provided 737 urine samples. Data for dietary habits and iodine status were available for 664 participants. Median urinary iodine excretion was 80.1 MUg/L (IQR 56.9-109.0). Urinary iodine measurements indicative of mild iodine deficiency were present in 51% (n=379) of participants, moderate deficiency in 16% (n=120), and severe deficiency in 1% (n=8). Prevalence of iodine deficiency was highest in Belfast (85%, n=135). Tap water iodine concentrations were low or undetectable and were not positively associated with urinary iodine concentrations. Multivariable general linear model analysis confirmed independent associations between low urinary iodine excretion and sampling in summer (p<0.0001), UK geographical location (p<0.0001), low intake of milk (p=0.03), and high intake of eggs (p=0.02). INTERPRETATION: Our findings suggest that the UK is iodine deficient. Since developing fetuses are the most susceptible to adverse effects of iodine deficiency and even mild perturbations of maternal and fetal thyroid function have an effect on neurodevelopment, these findings are of potential major public health importance. This study has drawn attention to an urgent need for a comprehensive investigation of UK iodine status and implementation of evidence based recommendations for iodine supplementation. FUNDING: Clinical Endocrinology Trust. PMID- 21640374 TI - Kynurenine 3-monooxygenase inhibition in blood ameliorates neurodegeneration. AB - Metabolites in the kynurenine pathway, generated by tryptophan degradation, are thought to play an important role in neurodegenerative disorders, including Alzheimer's and Huntington's diseases. In these disorders, glutamate receptor mediated excitotoxicity and free radical formation have been correlated with decreased levels of the neuroprotective metabolite kynurenic acid. Here, we describe the synthesis and characterization of JM6, a small-molecule prodrug inhibitor of kynurenine 3-monooxygenase (KMO). Chronic oral administration of JM6 inhibits KMO in the blood, increasing kynurenic acid levels and reducing extracellular glutamate in the brain. In a transgenic mouse model of Alzheimer's disease, JM6 prevents spatial memory deficits, anxiety-related behavior, and synaptic loss. JM6 also extends life span, prevents synaptic loss, and decreases microglial activation in a mouse model of Huntington's disease. These findings support a critical link between tryptophan metabolism in the blood and neurodegeneration, and they provide a foundation for treatment of neurodegenerative diseases. PMID- 21640376 TI - Iodine nutrition in the UK: what went wrong? PMID- 21640377 TI - The suppression of prostate LNCaP cancer cells growth by Selenium nanoparticles through Akt/Mdm2/AR controlled apoptosis. AB - The trace element Selenium is suggested having cancer prevention activity and used as food supplement. Previous results had shown Selenium nanoparticles are safer compared with other Selenium compounds like selenomethionine, sodium selenite and monomethylated Selenium, however, its anticancer activity and intrinsic mechanisms are still elusive. Here, we prepared Selenium nanoparticles and investigated its inherent anticancer mechanisms. We found Selenium nanoparticles inhibit growth of prostate LNCaP cancer cells partially through caspases mediated apoptosis. Selenium nanoparticles suppress transcriptional activity of androgen receptor via down-regulating its mRNA and protein expression. Moreover, Selenium nanoparticles activate Akt kinase by increasing its phosphorylation, promote Akt-dependent androgen receptor phosphorylation and Mdm2 regulated degradation through proteasome pathway. We suggest Selenium nanoparticles suppress prostate cancer cells growth by disrupting androgen receptor, implicating a potential application in cancer treatment. PMID- 21640378 TI - The use of nanoimprinted scaffolds as 3D culture models to facilitate spontaneous tumor cell migration and well-regulated spheroid formation. AB - Two-dimensional (2D) cell cultures are essential for drug development and tumor research. However, the limitations of 2D cultures are widely recognized, and a better technique is needed. Recent studies have indicated that a strong physical contact between cells and 2D substrates induces cellular characteristics that differ from those of tumors growing in vivo. 3D cell cultures using various substrates are then developing; nevertheless, conventional approaches have failed in maintenance of cellular proliferation and viability, uniformity, reproducibility, and/or simplicity of these assays. Here, we developed a 3D culture system with inorganic nanoscale scaffolding using nanoimprinting technology (nano-culture plates), which reproduced the characteristics of tumor cells growing in vivo. Diminished cell-to-substrate physical contact facilitated spontaneous tumor cell migration, intercellular adhesion, and multi-cellular 3D spheroid formation while maintaining cellular proliferation and viability. The resulting multi-cellular spheroids formed hypoxic core regions similar to tumors growing in vivo. This technology allows creating uniform and highly-reproducible 3D cultures, which is easily applicable for microscopic and spectrophotometric assays, which can be used for high-throughput/high-content screening of anticancer drugs and should accelerate discovery of more effective anticancer therapies. PMID- 21640379 TI - Effects of chronic treatment with fluoxetine on receptor-stimulated increase of [Ca2+]i in astrocytes mimic those of acute inhibition of TRPC1 channel activity. AB - Primary cultures of mouse astrocytes were used to investigate effects by chronic treatment (3-21 days) with fluoxetine (0.5-10 MUM) on capacitative Ca(2+) influx after treatment with the SERCA inhibitor thapsigargin and on receptor agonist induced increases in free cytosolic Ca(2+) concentration [Ca(2+)](i), determined with Fura-2. The agonists were the 5-HT(2B) agonist fluoxetine, the alpha(2) adrenergic agonist dexmedetomidine, and ryanodine receptor (RyR) and IP(3) receptor (IP(3)R) agonists. In untreated sister cultures each agonist distinctly increased [Ca(2+)](i), but in cultures treated for sufficient length of time or with sufficiently high doses of fluoxetine, acute administration of fluoxetine, dexmedetomidine, or RyR or IP(3)R agonists elicited reduced, in some cases abolished, effects. Capacitative Ca(2+) entry, meditated by TRPC1 channels, was sufficiently inhibited to cause a depletion of Ca(2+) stores, which could explain the reduced agonist effects. All effects of chronic fluoxetine administration could be replicated by TRPC1 channel antibody or siRNA. Since increases in astrocytic [Ca(2+)](i) regulate release of gliotransmitters, these effects may have profound effects on brain function. They may be important for therapeutic effects of all 5 conventional 'serotonin-specific reuptake inhibitors' (SSRIs), which at concentrations used therapeutically (~1 MUM) share other of fluoxetine's chronic effects (Zhang et al., Neuron Glia Biol. 16 (2010) 1-13). PMID- 21640380 TI - Identification of a novel, tissue-specific ABCG2 promoter expressed in pediatric acute megakaryoblastic leukemia. AB - ABCG2 encodes a transporter protein that is associated with multidrug-resistant phenotypes in many cancers, including acute myeloid leukemia (AML); high levels of expression are generally associated with a poor prognosis. To better understand how expression of ABCG2 is controlled in pediatric AML, we performed a detailed analysis of the ABCG2 transcript isoforms from a variety of tissue sources, including 85 pediatric AML samples. These studies revealed a complex 5' untranslated region (UTR) with 6 novel exons and multiple splice variants. Samples from children with acute megakaryoblastic leukemia (AML FAB-M7) not associated with Down syndrome showed uniformly higher levels of ABCG2 transcripts than samples from children with other AML subtypes. A novel 5' UTR identified 90kb upstream of the exon 2 translation initiation site was expressed only in M7 AML subtypes. An associated upstream promoter fragment was shown to be selectively expressed in megakaryoblastic leukemia cells but not in human epithelial cell lines. These findings identify a new tissue-specific ABCG2 promoter that is selectively expressed in pediatric M7 AML. We also show a relatively high incidence of ABCG2 mRNA expression in non-Down associated M7 AML, which may contribute to the relatively poor prognosis of the M7 AML subtype. PMID- 21640381 TI - Carbon monoxide, a reaction product of heme oxygenase-1, suppresses the expression of C-reactive protein by endoplasmic reticulum stress through modulation of the unfolded protein response. AB - The expression of C-reactive protein (CRP) rises rapidly in response to inflammation. The endoplasmic reticulum (ER) stress has been reported to cause CRP expression. Carbon monoxide (CO), a reaction product of heme oxygenase, exerts anti-inflammatory effects. In this study, we aimed to examine the role of CO in modulating ER stress-induced CRP expression. In HepG2 cells, ER stress triggered by tunicamycin, thapsigargin and homocysteine markedly induced CRP expression and the activation of protein kinase R-like endoplasmic reticulum kinase (PERK), inositol-requiring transmembrane kinase/endonuclease 1alpha (IRE1alpha), activating transcription factor 6 (ATF6), and hepatocyte-specific cyclic AMP response element binding protein H (CREBH). A CO-releasing molecule (CORM) inhibited ER stress-induced CRP expression. While CORM attenuated ER stress-induced activation of IRE1alpha, ATF6 and CREBH, it augmented PERK activation, which was associated with its inhibition of CRP expression. CORM also inhibited CRP expression in response to the pro-inflammatory cytokine IL-6 that was found to induce ER stress response in HepG2 cells. Moreover, in mice treated with the ER stress inducer tunicamycin, CORM administration reduced serum levels of CRP and the expression of CRP mRNA in the liver. Collectively, our findings suggest that CO may attenuate ER stress-induced CRP expression through modulation of the unfolded protein response. PMID- 21640382 TI - Severe adverse events after cataract surgery among medicare beneficiaries. AB - PURPOSE: To determine rates and risk factors associated with severe postoperative complications after cataract surgery and whether they have been changing over the past decade. DESIGN: Retrospective longitudinal cohort study. PARTICIPANTS: A total of 221 594 Medicare beneficiaries who underwent cataract surgery during 1994-2006. METHODS: Beneficiaries were stratified into 3 cohorts: those who underwent initial cataract surgery during 1994-1995, 1999-2000, or 2005-2006. One year rates of postoperative severe adverse events (endophthalmitis, suprachoroidal hemorrhage, retinal detachment) were determined for each cohort. Cox regression analyses determined the hazard of developing severe adverse events for each cohort with adjustment for demographic factors, ocular and medical conditions, and surgeon case-mix. MAIN OUTCOME MEASURES: Time period rates of development of severe postoperative adverse events. RESULTS: Among the 221 594 individuals who underwent cataract surgery, 0.5% (1086) had at least 1 severe postoperative complication. After adjustment for confounders, individuals who underwent cataract surgery during 1994-1995 had a 21% increased hazard of being diagnosed with a severe postoperative complication (hazard ratio [HR] 1.21; 95% confidence interval [CI], 1.05-1.41) relative to individuals who underwent cataract surgery during 2005-2006. Those who underwent cataract surgery during 1999-2000 had a 20% increased hazard of experiencing a severe complication (HR 1.20; 95% CI, 1.04-1.39) relative to the 2005-2006 cohort. Risk factors associated with severe adverse events include a prior diagnosis of proliferative diabetic retinopathy (HR 1.62; 95% CI, 1.07-2.45) and cataract surgery combined with another intraocular surgical procedure on the same day (HR 2.51; 95% CI, 2.07-3.04). Individuals receiving surgery by surgeons with the case-mix least prone to developing a severe adverse event (HR 0.52; 95% CI, 0.44-0.62) had a 48% reduced hazard of a severe adverse event relative to recipients of cataract surgery performed by surgeons with the case-mix most prone to developing such outcomes. CONCLUSIONS: Rates of sight-threatening adverse events after cataract surgery declined during 1994-2006. Future efforts should be directed to identifying ways to reduce severe adverse events in high-risk groups. PMID- 21640383 TI - Cosmesis after pterygium extended removal followed by extended conjunctival transplant as assessed by a new, web-based grading system. AB - OBJECTIVE: To design a web-based system to grade the cosmetic results after pterygium surgery and to use this to assess the aesthetic results of pterygium extended removal followed by extended conjunctival transplant. DESIGN: A standardized grading system with gradings of normal, excellent, good, fair, poor, and ungradeable was designed and tested by the author (standard gradings). This was then installed on a website where ophthalmologists and lay people could undertake grading and anatomy tutorials, and subsequently grade randomly assigned images in a masked fashion, of eyes after pterygium surgery and control eyes. PARTICIPANTS AND CONTROLS: A sample of 119 postoperative eyes were randomly selected from 279 consecutive surgeries together with 119 control eyes from the contralateral eyes of these patients supplemented with otherwise normal eyes. INTERVENTIONS: The author was tested twice on a sample of 40 images and then the full set, with 24 images repeated giving a total of 288 images using the proposed grading system, resulting in an intraobserver reliability score of 0.98. These images were then randomly presented to the graders, of whom 6 were postpterygium patients and 6 were corneal specialist ophthalmologists after they had completed the tutorials and passed a trial grading test. MAIN OUTCOME MEASURES: The percentage of images of postoperative and controls that were graded in each grading category were obtained and compared with the only existent study of post pterygium cosmetic results. RESULTS: Six lay people and 6 corneal specialists successfully passed the trial grading test of 40 images with weighted kappa of 0.70 to 0.85 and intraobserver reliability scores of 0.86 to 0.95 for lay graders and 0.90 to 0.92 for ophthalmic graders. Ninety-four percent of all pterygium surgery eyes were graded as acceptable; both ophthalmic and lay graders were unable to distinguish between postoperative and control eyes. CONCLUSIONS: This grading system is robust and user friendly, and pterygium extended removal followed by extended conjunctival transplant provides a very pleasing aesthetic result in most patients. PMID- 21640384 TI - Hepatitis B genotypes/subgenotypes and MHR variants among Moroccan chronic carriers. AB - OBJECTIVES: The aim of this study was to determine the prevalence of Hepatitis B Virus (HBV) genotypes, subgenotypes, HBV surface antigen (HBsAg) subtypes and naturally occurring mutations in Major Hydrophilic region (MHR) of HBsAg among Moroccan patients with chronic HBV infection. METHODS: The study included 200 patients chronically infected with HBV. The HBV genotypes, subgenotypes, HBsAg subtypes and MHR variants were determined by direct sequencing of the HBV surface (S) gene and phylogenetic analysis. RESULTS: The S gene was successfully amplified in 134 patients. The mean age was 40.6 +/- 12.2 years. Genotype D was predominant (90%, 120/134) and genotype A was less frequent (10%, 14/134). Genotype D strains belonged to subgenotypes D7 (70.8%, 85/120), D1 (25.8%, 31/120) and D2 (0.9%, 1/120). Three strains (2.5%) could not be classified in any subgenotype of genotype D. All genotype A strains belonged to subgenotype A2. HBsAg subtypes found were ayw2 (82.1%, 110/134), adw2 (10.4%, 14/134), ayw3 (3%, 4/134) and ayw4 (3%, 4/134). The global prevalence of MHR variants was 15% (20/134) with substitution P120T/S the most frequent (3.7%, 5/134). The occurrence of MHR variants was significantly associated with advancing age (>40 years) (p = 0.003) and independent of sex, HBeAg status, viral load, genotype, subgenotype and HBsAg subtype. CONCLUSIONS: This study provides the first description of predominance of HBV subgenotype D7/subtype ayw2 among Moroccan HBV chronic carriers. It also showed a significant prevalence of naturally occurring MHR variants in Morocco. PMID- 21640385 TI - Early warning signs checklists for relapse in bipolar depression and mania: utility, reliability and validity. AB - BACKGROUND: Recognising early warning signs (EWS) of mood changes is a key part of many effective interventions for people with Bipolar Disorder (BD). This study describes the development of valid and reliable checklists required to assess these signs of depression and mania. METHODS: Checklists of EWS based on previous research and participant feedback were designed for depression and mania and compared with spontaneous reporting of EWS. Psychometric properties and utility were examined in 96 participants with BD. RESULTS: The majority of participants did not spontaneously monitor EWS regularly prior to use of the checklists. The checklists identified most spontaneously generated EWS and led to a ten fold increase in the identification of EWS for depression and an eight fold increase for mania. The scales were generally reliable over time and responses were not associated with current mood. Frequency of monitoring for EWS correlated positively with social and occupational functioning for depression (beta=3.80, p=0.015) and mania (beta=3.92, p=0.008). LIMITATIONS: The study is limited by a small sample size and the fact that raters were not blind to measures of mood and function. CONCLUSIONS: EWS checklists are useful and reliable clinical and research tools helping to generate enough EWS for an effective EWS intervention. PMID- 21640386 TI - Development of a scale to assess the diurnal impact of insomnia. AB - The objective of this study was to develop and validate a scale to assess the diurnal impact of insomnia. The Insomnia Diurnal Impact Scale (IDIS) comprises six items designed to evaluate the daytime effects of insomnia. The psychometric properties of the original scale were analysed in a sample of 172 students, while its ability to differentiate insomniacs and non-insomniacs (according to the International Classification of Diseases (ICD)-10 criteria) was examined in a sample of 79 psychiatric patients and 82 individuals from the community. The psychometric properties of the English version were then analysed in a sample of 44 English-speaking participants. The results showed the internal consistency coefficient to be very good (0.86), with test-retest reliability at 1month being 0.79. A single factor explained almost 60% of the variance. Correlation of the IDIS with other scales varied between moderate and high values. Sensitivity was 78% and specificity 57% in the community sample, while the corresponding figures for the psychiatric population were 83% and 63%. Cronbach's alpha coefficient for the English version reached a value of 0.93. These results indicate that the IDIS shows adequate reliability and validity with both general and psychiatric populations, and also that it can discriminate between the presence and absence of insomnia. The English version presents good preliminary results regarding item corrected total correlation and internal consistency. In conclusion, the IDIS appears to be a useful tool in the primary care and mental health contexts for assessing insomnia-related diurnal dysfunction. PMID- 21640387 TI - Confirmatory Factor Analysis of Borderline Personality Disorder symptoms based on two different interviews: the Structured Clinical Interview for DSM-IV Axis II Disorder and the Revised Diagnostic Interview for Borderlines. AB - The diagnostic criteria for Borderline Personality Disorder (BPD) are heterogeneous, and include an admixture of personality traits, behaviours, and symptoms. The BPD DSM factor structure has been extensively studied, even though results are not consistent. In this study we performed a confirmatory factor analysis (CFA) to compare the five-factor model reported by Oldham, a three factor model, and a unidimensional model of BPD diagnosis criteria. This study validates the findings of previous studies by performing a CFA with the DSM-IV BPD criteria and information derived from the DIB-R. A sample of 338 patients referred to our outpatient BPD program participated in the study. Results of the CFA support both the hypothesized unidimensional and the three-factor models, whereas the five-factor model was not confirmed. However, the three-factor model fits better than the unidimensional model. Thus, although the DSM-IV BPD criteria conceptualize BPD as a unidimensional structure, our results give support to the idea that the three-factor model could offer a better approach to further improve the current treatment of BPD, as well as lead to a better understanding of its ethiopathogenesis and comorbidity analysis. PMID- 21640388 TI - Immune response of cattle immunized with a conjugate of the glycolipid glucose monomycolate and protein. AB - Strong anti glycolipid IgG responses can occur in humans and animals, but contrary to anti protein responses and anti glycoprotein responses, the exact mechanism of induction is unknown. We have previously shown that experimental immunization with the glycolipid glucose monomycolate (GMM) causes the development of specific T cell responses, but not of anti GMM antibodies. However, cattle naturally infected with Mycobacterium avium ssp. paratuberculosis produce high levels of anti GMM IgG. In the present study, we tested whether vaccination with GMM conjugated to a protein mimics natural infection in its capacity to induce the production of antibodies against GMM. Cattle were immunized (n=5 per group) with GMM conjugated to a protein, or GMM and protein non-conjugated and administered at contralateral locations, or carrier only. Although immunization with the GMM-protein conjugate vaccine and the non conjugated vaccine induced protein specific antibody responses, GMM specific antibodies were not detected in either of the groups. In conclusion, the generation of isotype-switched anti lipid antibodies appears to require more than providing peptide epitopes for T helper cells to support glycolipid specific B cells in antibody production. PMID- 21640389 TI - Identification and expression profile of multiple genes in Nile tilapia in response to formalin killed Streptococcus iniae vaccination. AB - Twenty-eight expressed sequence tags (ESTs) were isolated from a Nile tilapia (Oreochromis niloticus) vaccinated vs non-vaccinated subtractive library at 12-h post injection of a formalin killed Streptococcus iniae ARS-98-60 vaccine. The 28 ESTs were classified in terms of their putative functions. Half of the ESTs identified were unknown proteins. Of the remaining half ESTs, 17% have putative functions in protein biosynthesis and 11% have putative functions in immunity, energy production, and signal transduction, respectively. Immunity-related ESTs identified included high density lipoprotein-binding protein vigilin, immunoglobulin heavy chain, and QM-like protein. Quantitative PCR revealed that one EST (cytochrome c oxidase subunit II) was highly upregulated (1825 +/- 336 fold) in vaccinated fish compared to that in non-vaccinated fish. Of the remaining 27 ESTs, nine were significantly (P<0.05) upregulated (<20 fold) in vaccinated fish. The nine significantly upregulated genes included five unknown or hypothetical proteins and four known proteins (high density lipoprotein binding protein vigilin, QM-like protein, ribosomal protein S13, and ribosomal protein L5). The upregulation of these genes induced by killed S. iniae vaccines suggest that they might play important role in Nile tilapia defense against S. iniae infection. PMID- 21640390 TI - Pathophysiology of experimental Aeromonas hydrophila infection in Puntius sarana: early changes in blood and aspects of the innate immune-related gene expression in survivors. AB - A decline in production of Puntius sarana worldwide and subsequent intensification of its culture as a measure for conservation and increase food fish production have led to emergence of diseases. Investigations on innate immunological changes in fish that survives an acute bacterial infection are a few or lacking. An attempt was made to study the detailed changes in innate immune responses in P. sarana following an intraperitoneal challenge with Aeromonas hydrophila at 2.24 * 10(7)colony-forming unit (CFU)/fish. Sequential changes in transcription of immune related genes in liver and kidney, innate immune status, haematological profile and histopathology of major organs were studied after different time intervals post-challenge in the survivors. A significant increase in serum myeloperoxidase, ceruloplasmin activities and total leucocyte count, and decline in alternative complement activity were observed at early infection periods. A significant decline in plasma glucose level, total erythrocyte count, haemoglobin content and packed cell volume in blood was noticed after challenge. Superoxide radical production and serum antiprotease activity increased significantly at later period of challenge. In addition, few immune related genes pertinent to acute phase reactants and inflammatory response viz., complement component 3 (C3), lysozyme G, lysozyme C, beta2-microglobulin (B2M), transferrin, interleukin 1beta, interleukin 8, manganese superoxide dismutase (MnSOD), natural killer cell enhancing factor-B and chemokine CXCa were modulated after bacterial challenge. In liver, a significant up-regulation of C3 and transferrin, and down-regulation of lysozyme G, interleukin 8, MnSOD and B2M were noticed in survivors. On the contrary, lysozyme G, lysozyme C, interleukin 1beta, interleukin 8, CXCa, and MnSOD were found to be significantly up-regulated and C3 was down-regulated in the kidney. All these data suggest that the innate immune factors were highly modulated during infection process to render protection of P. sarana to this bacterium. This study also generated the primary information on many innate immune molecules/genes for this important medium carp species for the first time. PMID- 21640391 TI - Upregulation of microglial C1q expression has no effects on nigrostriatal dopaminergic injury in the MPTP mouse model of Parkinson disease. AB - Here we analyzed C1q, the initial recognition subcomponent of classical complement activation cascade, in an experimental model of Parkinson disease (PD). Nigrostriatal dopaminergic pathway injury was induced by treatment of wildtype mice subchronically with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Constitutive expression of C1q was restricted to microglia throughout the brain, and microglial C1q expression was early and transiently upregulated after MPTP in the substantia nigra (SN) and striatum, as analyzed by immunohistochemistry and in situ hybridization. C1q-positive microglia exhibited morphological characteristics of activated macrophage-type of cells, co-stained for MHCII, proliferated and were in close contact with degenerating dopaminergic neurons and fibers in the MPTP-lesioned SN. However, mice deficient in functional C1q protein were not significantly different in MPTP-induced loss of nigral dopaminergic neurons, striatal dopaminergic fibers and dopamine levels than their control littermates. In conclusion, C1q is upregulated and considered to be a marker of microglial activation in the nigrostriatal system after subchronic MPTP, but nigrostriatal dopaminergic injury may be not affected by C1q in this model. PMID- 21640392 TI - Immunosuppressive activity of a novel peptide analog of alpha-melanocyte stimulating hormone (alpha-MSH) in experimental autoimmune uveitis. AB - Autoimmune uveitis is an inflammatory disorder of the eye that can lead to pain and vision loss. Steroids and immunosuppressive drugs are currently the only therapeutics for uveitis and have serious ocular and systemic toxicities. Therefore, safer alternative therapeutics are desired. Alpha-melanocyte stimulating hormone (alpha-MSH) is a neuropeptide that suppresses effector T cell functions, induces regulatory T cells and has beneficial effects in certain autoimmune and transplant models. A novel d-amino acid peptide analog of native alpha-MSH (dRI-alpha-MSH) was produced that was protected from protease digestion and had increased selectivity for the melanocortin-1 receptor. Systemic delivery of the dRI-alpha-MSH analog dramatically suppressed disease progression and retained retinal architecture in the experimental autoimmune uveitis (EAU) model. Local delivery by periorbital injection was equally effective. Importantly, treatment with the novel dRI-alpha-MSH analog suppressed uveitis with a similar magnitude to the corticosteroid, dexamethasone. Data indicate that the novel dRI alpha-MSH analogs show anti-inflammatory activities and have potential therapeutic use in uveitis and other autoimmune diseases. PMID- 21640393 TI - Pediatric obstructive fibrinous tracheal pseudomembrane--characteristics and management with flexible bronchoscopy. AB - OBJECTIVE: To evaluate the incidence, characteristics, flexible bronchoscopy (FB) findings, interventions and outcome of pediatric obstructive fibrinous tracheal pseudomembrane (OFTP) in our pediatric and neonatal intensive care units (ICUs). PATIENTS AND METHODS: This is a retrospective study of medical and FB video records in a single tertiary university-affiliated teaching hospital over a ten year period. Data was collected from patients who were admitted and extubated of endotracheal tube in the ICUs with a FB diagnosis of an OFTP-like lesion. The associated medical information, FB interventions and video records were reviewed and analyzed. RESULTS: Eight patients with OFTP were enrolled, with an incidence rate of 1.48% in the postextubation respiratory distress patients. Mean age was 32 +/- 32 months (range, 2 months to 13 years); mean body weight was 13.7 +/- 8.1 kg (range, 4.3-45 kg); mean intubation period was 37.6 +/- 12.3h; mean time for symptoms to develop after extubation was 3.6 +/- 1.4h. Symptoms lasted for 20.8 +/- 20.3h before FB examination. All patients were accurately diagnosed with OFTP at the first postextubation FB examination and revealed various morphologies. The estimated cross-sectional tracheal lumen was reduced by 70-90% and the mean length of lesion was 18.1 +/- 5.2mm (range, 10-30 mm). All OFTP were successfully ablated immediately after the diagnosis in one FB session by using various techniques and without any complication. Total duration for both diagnostic and interventional FB was 19.4 +/- 2.5 min. No recurrence was noted thereafter. CONCLUSIONS: OFTP should always be considered in the event of postextubation respiratory distress, especially in the pediatric and neonatal ICUs. Early diagnosis and effective ablation can be achieved with aid of FB. PMID- 21640394 TI - Health-related factors associated with hospitalization for old people: comparisons of elderly aged 85 in a population cohort study. AB - The aim of this population-based study was to (1) describe living conditions and actual health care utilization among 85 year olds; (2) determine factors that affect hospital admissions in this age. The study was conducted on 85-year-old residents in Linkoping municipality, Sweden. The data collected included medical records, health care utilization during the preceding 12 months and a postal questionnaire on assistance, assistive technology, functional impairment, feelings of loneliness, worries and health-related quality of life measured by the EQ-5D. Out of 650 eligible individuals, 496 (78% of those alive) participated. Despite the prevalence of multi-morbidity (68%) and mental discomfort, the majority managed self-care (85%), usual activities (74%) and had high (>60/100) self-rated health evaluated by a visual analog scale (VAS). The non-hospitalized group reported a better health status than the hospitalized group in terms of medical aspects, living conditions and subjective estimation. Factors associated with in-patient care were an increased number of general practitioner visits, more assistive technology, community assistance, multimorbidity and/or diagnosed congestive heart failure and arrhythmia. PMID- 21640395 TI - Healthy centenarians show high levels of circulating interleukin-22 (IL-22). AB - Aging is characterized by a progressive alteration of homeostatic mechanisms modulated by environmental and genetic factors. It is associated with a pro inflammatory status. In centenarians, an increase of pro-inflammatory cytokine production balanced by anti-inflammatory immune response that would promote longevity is observed. Cytokine dysregulation is believed to play a key role in the proposed remodeling of the immune-inflammatory responses accompanying old age. IL-22 is a pro-inflammatory cytokine belonging to the IL-10 family and represents an important effector molecule of activated T helper (Th)-22, Th-1, and Th-17 cells. We recruited 17 healthy centenarians (4 males, 13 females, range 100-105 years). All ultralongeval subjects were living at home or in a nursing home. Sixteen healthy, sex-matched individuals (4 males, 12 females, range 60-95 years) were also recruited as controls. Centenarians displayed significantly higher circulating IL-22 levels compared to control population (45.7+/-66.9 pg/ml versus 11.1+/-6.5 pg/ml; p=0.031). It's well known that IL-22 is a pro inflammatory cytokine produced by activated T lymphocytes and NK cells. IL-22 stimulates the production of acute phase reactants and promotes the antimicrobial defense. The results of the present study show, for the first time, that there is an increase of IL-22 in healthy centenarians. This pro-inflammatory condition probably is protective against infection, promoting the longevity of these subjects. PMID- 21640396 TI - Hypothyroidism in the elderly: diagnostic pitfalls illustrated by a case report. AB - A diagnosis of hypothyroidism in the elderly can easily be overlooked if we rely exclusively on its clinical presentation because this may be highly non-specific, since the signs and symptoms of the disease are common to other diseases typical of old age, and even to the normal aging process. Imaging diagnostics (ultrasound or CT), when considered alone, are also of little use for the purpose of clarifying thyroid gland function. We report here on a case of primary hypothyroidism that was diagnosed late because the correlated signs and symptoms (asthenia, bradycardia, pleural effusions, hyponatremia, worsening renal and respiratory insufficiency, hoarseness) had previously been attributed to the normal aging process and to the patient's other health conditions (Parkinson's disease, PD; chronic obstructive pulmonary disease, COPD). After a couple of weeks of treatment with levothyroxine and liothyronine, there were clinical and laboratory evidences of an improvement in the patient's condition. She became more reactive, with a shriller voice. The pleural effusion disappeared, and so did the bradycardia. Laboratory tests showed normal sodium levels, and the renal insufficiency had improved. The lack of specificity of the clinical presentation of hypothyroidism in the elderly might justify the routine measurement of thyroid stimulating hormone in these patients. PMID- 21640397 TI - Age, conduction defects and restrictive lung disease independently predict cardiac events and death in myotonic dystrophy. AB - OBJECTIVE: The aim of the study was to identify, in addition to conduction defects, possible predictors of cardiac events and death in patients with myotonic dystrophy (DM1). METHODS AND DESIGN: A retrospective observational cohort study was undertaken. Baseline clinical and non-invasive cardiac and respiratory investigations were obtained from 107 DM1 patients, who were regularly re-examined. Primary end-points were occurrence of cardiac events (pacemaker implantation or tachyarrhythmia) or death. Probability of an event was calculated using the Kaplan-Meier method, while contributing factors were assessed using univariate and multivariate (Cox model) analyses. RESULTS: Cardiac events occurred in 34 patients (29%). Age, muscular impairment, infantile onset, restrictive lung disease (RLD), ECG conduction defects, left ventricular ejection fraction (LVEF) below 50%, and arrhythmia detected during Holter monitoring were predictors of cardiac events. Multivariate analysis indicated that age, RLD, ECG conduction defects, Holter arrhythmia and LVEF remained independent predictors. Probability of cardiac events was 2.5% (5%CI: 0-7%) at 1 year and 6% (5%CI: 0 14%) at 3 years in patients younger than 42 years with normal ECG, Holter, LVEF and lung volumes. Advancing age, distal or proximal weakness and RLD characterized all non-survivors (n=14). CONCLUSION: Cardiac events or death are predicted not only by conduction defects or cardiomyopathy in DM1, but also by RLD, muscular disability and advancing age. Addition of these criteria should modulate time intervals for patient follow-up examinations. In young patients with normal baseline investigations, screening investigations every 2 or 3 years seem to be sufficient. PMID- 21640398 TI - Correlation of NT-proBNP, proANP and novel biomarkers: copeptin and proadrenomedullin with LVEF and NYHA in patients with ischemic CHF, non-ischemic CHF and arterial hypertension. PMID- 21640399 TI - Regression of atherosclerosis plaques in apolipoprotein E-/- mice after lentivirus-mediated RNA interference of CD40. AB - BACKGROUND: A role of CD40 (cluster of differentiation 40) is suggested in development of atherosclerosis plaques, especially in advanced plaques. However, the role of lentiviruses carrying small interfering RNA (siRNA) of CD40 in progression and destabilization of advanced atherosclerotic plaques remains unknown. The aim of this study was to determine whether inhibition of CD40 signaling by lentivirus-mediated RNA interference (RNAi) could inhibit progression of atherosclerotic plaques and increase collagen production. METHODS: Apolipoprotein E-deficient (ApoE-/-) mice aged 10 weeks were fed a high-fat diet and a constrictive collar was placed around right carotid arteries of these mice to induce plaques formation. The recombinant CD40-RNAi-Lentivirus (CD40-RNAi-LV) or negative control-green fluorescent protein-Lentivirus (NC-GFP-LV) were constructed and transfected into right carotid plaques respectively eight weeks after surgery. RESULTS: CD40-RNAi-LV not only prevented plaques progression but also decreased plaques content of lipid, increased plaques content of collagen 6 weeks after lentivirus transfection. This effect reflected a marked decrease in the intima/media ratios (0.31 +/- 0.04 vs 0.68 +/- 0.05, P<0.05) and a diminished degree of lumen stenosis (intima/lumen ratios, 0.17 +/- 0.04 vs 0.33 +/- 0.40, P<0.05). Moreover, real-time polymerase chain reaction (RT-PCR) analysis of CD40 RNAi-LV group downregulated expressions of proinflammatory cytokines, chemokines and matrix metalloproteinases. CONCLUSIONS: Lentivirus-mediated CD40 silencing by siRNA treatment would be a new strategy to inhibit plaques progression and to reduce local inflammation through the antiinflammatory effects. PMID- 21640400 TI - Multi- versus single-vessel percutaneous coronary intervention for ST-elevation myocardial infarction with multi-vessel disease. PMID- 21640401 TI - An updated meta-analysis of randomized controlled trials of everolimus- versus paclitaxel-eluting stents. PMID- 21640402 TI - Transcatheter intervention following bosentan-iloprost treatment in a patient with patent ductus arteriosus and pulmonary hypertension. PMID- 21640403 TI - Cardiac magnetic resonance can early assess the presence and severity of heart involvement in Naxos disease. PMID- 21640404 TI - Contrast transthoracic echocardiography versus transcranial Doppler for patent foramen ovale detection. PMID- 21640405 TI - Adipose tissue-derived stem cells embedded with eNOS restore cardiac function in acute myocardial infarction model. AB - This study assessed the potential therapeutic efficacy of endothelial NO syntheses (eNOS)-expressing adipose tissue-derived stem cells (ADSCs) on infarcted hearts. We isolated CD29+, CD44+, CD45- cells from adipose tissue. Multipotent property of ADSCs was characterized by induction to differentiate into myogenic, neurogenic, and endothelic lineages. We hypothesized that combination of eNOS over-expression and transplantation of ADSCs could restore NO bioavailability and improve cardiac function in infarcted hearts. Here with several lines of experimental evidences, we demonstrated that ADSCs with eNOS overexpression induced eNOS expression in host endothelial cells and vascular smooth muscle cells, both in vitro and in vivo. This effect was possibly mediated by calcium signal. Transplantation of ADSCs with eNOS embedded showed great therapeutic efficacy in reduction of infarcted size, compared with normal ADSC. Results of this study suggest that ADSCs could be an attractive vehicle for the exogenous eNOS expression into heart after infarction, which is beneficial to restoration of cardiac function. Paracrine effect by mobilizing the host endothelial cells and smooth muscle cells may be the mechanism underlying the therapeutic effect. PMID- 21640406 TI - Uhl's anomaly. PMID- 21640407 TI - Effect of transplantation with autologous bone marrow stem cells on acute myocardial infarction. AB - BACKGROUND: This study was designed to investigate the effects of treatment with autologous mesenchymal stem cells (MSCs) by intracoronary transplantation on myocardial infarction (MI) in swine. METHODS: MSCs were transfected with a lentiviral vector carrying the gene encoding green fluorescent protein (GFP) and labeled in vitro with superparamagnetic iron oxide (SPIO). An acute MI (AMI) model was established by percutaneous balloon occlusion. Dual-labeled MSC suspensions were injected through the infarct-related coronary artery using an over-the-wire (OTW) balloon device in the experimental group (n=5), while normal saline was injected into the control (n=5). Stem cell migration and improvements of cardiac function were evaluated by magnetic resonance imaging (MRI), single photon emission computed tomography (SPECT), and ultrasonic cardiogram (UCG) both before MSC transplantation and after 8 weeks. At 8weeks after transplantation, myocardial tissue was analyzed by histopathologic analyses. RESULTS: Blue SPIO particles were in the cytoplasm of the MSCs. The labeling efficiency reached 100%. MRI revealed hypointensities of SPIO-labeled MSCs that were clustered in the myocardial infarct area after MSC transplantation. As time progressed, the signal gradually weakened and the area shrank. SPECT revealed that the myocardial filling defect was reduced and the amount of surviving myocardium was significantly increased at 8weeks. UCG revealed significant improvement when compared with the control at 8 weeks after transplantation. Prussian blue positive cells were observed in the MI border zone. Fluorescence-positive cells were also observed, but the number of such cells was fewer than before. CONCLUSIONS: In vivo serial tracking of SPIO-labeled MSCs can be achieved by MRI. Intracoronary transplantation of SPIO-labeled MSCs can increase cardiac function and promote myocardial viability. PMID- 21640408 TI - Everolimus- versus first generation drug-eluting stents: the never-ending story of searching for a winner in the "limus family". PMID- 21640409 TI - Clonidine therapy in patients with heart failure improves exercise efficiency, functional class and symptoms. PMID- 21640410 TI - Elastic properties of the ascending aorta in patients with rheumatoid arthritis. PMID- 21640411 TI - Impaired mobilization of bone marrow derived CD34 positive mononuclear cells is related to the recurrence of atrial fibrillation after radiofrequency catheter ablation. AB - BACKGROUND: We have reported previously that non-ischemic titrated cardiac injury by radiofrequency catheter ablation (RFCA) of atrial fibrillation (AF) mobilizes bone marrow derived CD34+ mononuclear cells. We hypothesized that the degree of post-RFCA CD34+ cell mobilization affects the clinical outcome of AF ablation. METHODS AND RESULTS: Fifty-six patients (39 males, 53 +/- 13 years) who underwent electrophysiology study (EPS; n=10) or RFCA of AF (n=46) were included. The peripheral blood CD34+ cell count and multiple serologic markers were evaluated before, immediately after the procedure. RESULTS: 1. The percent increase of CD34+ cells (%DeltaCD34+) was significant after RFCA as compared to EPS (p < 0.01). 2. The post-RFCA CD34+ cell count was significantly higher in patients who underwent RF energy delivery >=80 min than those <80 min (p = 0.024). 3. The %DeltaCD34+ was linearly correlated with the plasma level of troponin I (R = 0.38, p < 0.01), but not with the non-ablation procedure time (p = NS). 3. During 30.2 +/- 2.7 months follow-up, AF recurred in 37% of patients including early recurrence (34.8%). In contrast, the patients in whom AF recurred received a longer duration of RF energy delivery than those remaining in sinus rhythm (p = 0.04), they were associated with lower %DeltaCD34+ (p = 0.02). CONCLUSION: CD34+ mononuclear cells were mobilized after catheter ablation by RF energy dose dependent manner, and the duration of RF energy delivery was longer in patients with AF recurrence. However, CD34+ mononuclear cell mobilization was significantly impaired in patients with recurring AF after RFCA. PMID- 21640412 TI - Ultrafiltration for acute decompensated heart failure: financial implications. AB - Heart failure is the leading cause of hospitalization in older patients and is considered a public health problem with a significant financial burden on the health care system. Ultrafiltration represents an emerging therapy for patients with heart failure with a number of advantages over the conventional therapy. In this article, a summary of the relevant pathophysiological mechanisms such as removal of inflammatory cytokines are provided that might indeed be associated with a number of financial implications for ultrafiltration. Then practical points such as training of physicians and staff that need to be considered by physicians and medical centers with regards to financial implications of this therapy are reviewed. PMID- 21640413 TI - Kounis syndrome during general anaesthesia and administration of adrenaline. PMID- 21640414 TI - A meta-analysis of first-generation drug-eluting vs bare-metal stents for coronary chronic total occlusion: effect of length of follow-up on clinical outcome. PMID- 21640415 TI - Veterinary syndromic surveillance: Current initiatives and potential for development. AB - This paper reviews recent progress in the development of syndromic surveillance systems for veterinary medicine. Peer-reviewed and grey literature were searched in order to identify surveillance systems that explicitly address outbreak detection based on systematic monitoring of animal population data, in any phase of implementation. The review found that developments in veterinary syndromic surveillance are focused not only on animal health, but also on the use of animals as sentinels for public health, representing a further step towards One Medicine. The main sources of information are clinical data from practitioners and laboratory data, but a number of other sources are being explored. Due to limitations inherent in the way data on animal health is collected, the development of veterinary syndromic surveillance initially focused on animal health data collection strategies, analyzing historical data for their potential to support systematic monitoring, or solving problems of data classification and integration. Systems based on passive notification or data transfers are now dealing with sustainability issues. Given the ongoing barriers in availability of data, diagnostic laboratories appear to provide the most readily available data sources for syndromic surveillance in animal health. As the bottlenecks around data source availability are overcome, the next challenge is consolidating data standards for data classification, promoting the integration of different animal health surveillance systems, and also the integration to public health surveillance. Moreover, the outputs of systems for systematic monitoring of animal health data must be directly connected to real-time decision support systems which are increasingly being used for disease management and control. PMID- 21640416 TI - Coxiella burnetii (Q fever) in Rattus norvegicus and Rattus rattus at livestock farms and urban locations in the Netherlands; could Rattus spp. represent reservoirs for (re)introduction? AB - The Q fever outbreak in the Netherlands in 2007-2010 prompted government interventions to reduce the human incidence by reduction of Q fever shedding at dairy goat farms. Mandatory hygiene measures were taken, including the control of animal reservoirs. It has been postulated that brown rats, through their commensal nature, form an important factor in the persistent dissemination of endemic circulating Coxiella burnetii in nature to domestic animals, livestock and humans. Here, the occurrence of C. burnetii in rats captured at different types of location during the Q fever outbreak in the Netherlands, viz. urban areas, nature areas and various types of farm has been determined. This is a first step towards the elucidation of the reservoir status of rats in veterinary and human Q fever epidemiology. C. burnetii DNA was detected in the spleen of 4.9% of the brown rats (Rattus norvegicus) and 3.0% of the black rats (Rattus rattus). Evidence for C. burnetii infection was also found in liver, kidney, lung and intestinal tissue but not in heart, brain and pancreas. C. burnetii IgGs were detected in 15.8% of the brown rats. Positive rats were collected at goat, pig, cattle and poultry farms, and urban locations; including locations outside the designated 5km "increased-risk" zones around bulk milk positive goat farms. The percentage of rat-positive locations was the highest for goat farms (50%) and cattle farms (14.3%). The presence of actively infected rats outside the lambing season and at multiple environmental settings including urban locations might suggest that rats are not merely a spill-over host due to infection by a contaminated environment but might represent true reservoirs, capable of independent maintenance of C. burnetii infection cycles and thereby contributing to spread and transmission of the pathogen. If frequent (re)introduction of C. burnetii to small ruminant farms can be caused by rats as maintenance reservoirs, mandatory wildlife control and lifelong vaccination of herds will be necessary. PMID- 21640417 TI - Expression of industrially relevant laccases: prokaryotic style. AB - Laccases are a class of multi-copper oxidases (MCOs) that catalyze the one electron oxidation of four equivalents of a reducing substrate, with the concomitant four-electron reduction of dioxygen to water. They can catalyze a multitude of reactions, including the degradation of polymers and oxidative coupling of phenolic compounds, positioning them as significant industrial enzymes. Although fungal laccases are well known and well characterized, only recently has in silico biology led to rapid advances in the discovery, characterization and engineered expression of prokaryotic laccases. We describe the recent burgeoning of prokaryotic laccases, their catalytic properties, structural features and molecular evolution, vis-a-vis fungal laccases where possible. Special focus is given to the application of laccases to the emerging cellulosic biofuel industry. PMID- 21640418 TI - Vaccine design of hemagglutinin glycoprotein against influenza. AB - Influenza viruses continue to cause annual epidemics and pose the threat of a deadly global pandemic. Vaccination has remained the best approach for prevention and control of influenza infection. However, current influenza vaccines are only effective against closely-matched circulating strains, and therefore must be updated and administered every year. In this review, we discuss recent developments in the search for better influenza vaccines, especially using the major virus surface glycoprotein hemagglutinins (HAs). Understanding how glycans on HAs affect the immune response and knowledge of how broadly neutralizing antibodies are induced will pave the way for a cross-protective influenza vaccine that does not require frequent updates or annual immunizations. PMID- 21640419 TI - An increased utilisation rate and better compliance to guidelines for primary radiotherapy for breast cancer from 1997 till 2008: a population-based study in the Netherlands. AB - Only scarce data are available on the utilisation rate of primary radiotherapy (RT) for patients with breast cancer. In this study, we compared the use of primary RT for patients with stages I-III breast cancer in 4 of the 9 Dutch Comprehensive Cancer Centres, focussing specifically on time trends as well as age effects. From the population-based cancer registries, we selected all females diagnosed with breast cancer between 1997 and 2008 (N=65,966, about 50% of all Dutch breast cancer patients in this period). We observed an overall increase in the use of primary RT for breast cancer patients ranging from 55-61% in 1997 to 58-68% in 2008. This can be explained by a higher rate of breast-conserving surgery (BCS), which was followed by RT in 87-99% of cases, and a reduced rate of total mastectomy (TM) which was followed by RT in 26-47% of cases. Increasing age was associated with a reduced use of RT, especially for those above 75. Finally, we observed a decrease in time of observed regional variances in the use of RT after BCS as well as after TM (for stage III disease). These findings can be attributed to the development and implementation of the Dutch nationwide guidelines for treatment of breast cancer. PMID- 21640420 TI - Correlation between the serum KL-6 level and the grade of radiation pneumonitis after stereotactic body radiotherapy for stage I lung cancer or small lung metastasis. PMID- 21640421 TI - Re-irradiation with scanned charged particle beams in recurrent tumours of the head and neck: acute toxicity and feasibility. AB - BACKGROUND: Treatment of surgically unresectable recurrence in the head and neck region remains a therapeutic problem with the only curative option being a second course of radiation with a tumouricidal dose. We report initial toxicity and efficacy of charged particle therapy in this situation. METHODS: Treatment related side-effects of patients treated with charged particle beams for recurrent tumours of the head and neck were prospectively collected and patient data was retrospectively analysed with regard to toxicity and efficacy of the treatment according to CTCAE v. 4.03 and RECIST. RESULTS: Treatment was tolerated well without any severe acute toxicity. In non-chordoma/chondrosarcoma patients, overall response rate was 53.3% at 8 weeks post RT. 4/5 chordoma/chondrosarcoma patients showed no signs of further tumour progression. CONCLUSION: Initial experience of re-irradiation with scanned particle beams in recurrent tumours of the head and neck seems feasible and encouraging. Further follow-up is needed to investigate potential late effects. PMID- 21640422 TI - Three-dimensional conformal hypofractionated simultaneous integrated boost in breast conserving therapy: results on local control and survival. AB - PURPOSE: To report on local control and survival after breast conserving therapy (BCT) including three-dimensional conformal simultaneous integrated boost irradiation (3D-CRT-SIB) and on the influence of age on outcome. PATIENT AND METHODS: For this study, 752 consecutive female breast cancer patients (stages I III), treated with 3D-CRT-SIB at the University Medical Center Groningen from 2005 to 2008, were retrospectively identified. Median age was 58.4 (range 26-84) years. The SIB fractionation used was: 28*1.8Gy (whole breast) and 28*2.3Gy or 2.4Gy (tumour bed). Next to outcome, we estimated the effect of age on the recurrence-free period (RFP) by multivariate Cox regression survival analysis. RESULTS: Median follow-up was 41 (range 3-65) months. Local control was 99.6% at 3 years (6 ipsilateral recurrences). The 3-year locoregional control, RFP and overall survival (OS) rates were 99.2%, 95.5%, and 97.1%, respectively. In multivariate analysis, tumours >2cm (hazard ratio (HR) 3.11; 95% confidence interval (CI) 1.57-6.17) and triple negativity (HR 3.03; 95% CI 1.37-6.67) and not age were associated with impaired RFP. CONCLUSIONS: At 3 years, the 3D-CRT SIB technique in BCT results in excellent local control and OS. Age was not a risk factor for any recurrence. PMID- 21640423 TI - Treatment of primary liver cancer using highly-conformal radiotherapy with kV image guidance and respiratory control. AB - PURPOSE: To implement a reliable, practical and reproducible treatment procedure, based on in-room kV-image guidance and respiratory control, for liver cancer patients treated with high dose conformal radiotherapy using a commercially available treatment system. MATERIALS AND METHODS: CT stimulation was conducted under voluntary breath hold or gating using the Varian Real-time Position ManagementTM (RPM) System. Treatments were delivered daily under kV image guidance to verify the diaphragmatic or lipiodol-defined tumor position. RESULTS: Thirty-three patients with liver confined hepatocellular carcinoma were treated between May 2006 and Dec 2009. After a median follow-up period of 16.5 months (range: 3.5-40.7), all but 2 patients demonstrated radiological tumor regression. Eight patients (24%) achieved complete remission. The median tumor shrinkage was 42% (27-100%). Subsequent in-field tumor progression was observed in only three patients (10%). For the 23 patients with abnormal alpha fetoprotein level, 22 of them showed biochemical response with a median AFP level drop of 78%. The treatment was well tolerated: Grade 3 toxicities occurred in 5 patients (1 leucopenia, 1 elevated liver enzyme and 3 elevated bilirubin level) but there was no grade 4 toxicity or treatment related death. The 1 year overall survival rate is 71.7% and median survival time is 17.2 months (3.5-40.7 months). CONCLUSIONS: Excellent treatment results with minimal toxicities could be achieved in a clinical environment with a commercially available highly sophisticated radiotherapy system. PMID- 21640424 TI - Multifactorial intervention in individuals with type 2 diabetes and microalbuminuria: the Microalbuminuria Education and Medication Optimisation (MEMO) study. AB - AIMS: To determine whether tighter cardiovascular risk factor control with structured education in individuals with type 2 diabetes (T2DM) and microalbuminuria benefits cardiovascular risk factors. METHODS: Participants from a multiethnic population, recruited from primary care and specialist clinics were randomised to intensive intervention with structured patient (DESMOND model) education (n=94) or usual care by own health professional (n=95). PRIMARY OUTCOME: change in HbA1c at 18months. SECONDARY OUTCOMES: changes in blood pressure (BP), cholesterol, albuminuria, proportion reaching risk factor targets, modelled cardiovascular risk scores. RESULTS: Mean (SD) age and diabetes duration of participants were 61.5 (10.5) and 11.5 (9.3) years, respectively. At 18months, intensive intervention showed significant improvements in HbA1c (7.1(1.0) vs. 7.8(1.4)%, p<0.0001), systolic BP (129(16) vs. 139(17) mmHg, p<0.0001), diastolic BP (70(11) vs. 76(12) mmHg, p<0.001), total cholesterol (3.7(0.8) vs. 4.1(0.9) mmol/l, p=0.001). Moderate and severe hypoglycaemia was 11.2 vs. 29.0%; p=0.001 and 0 vs. 6.3%; p=0.07, respectively. More intensive participants achieved >=3 risk factor targets with greater reductions in cardiovascular risk scores. CONCLUSIONS: Intensive intervention showed greater improvements in metabolic control and cardiovascular risk profile with lower rates of moderate and severe hypoglycaemia. Intensive glycaemic interventions should be underpinned by structured education promoting self-management in T2DM. PMID- 21640425 TI - Strategies for viral RNA stability: live long and prosper. AB - Eukaryotic cells have a powerful RNA decay machinery that plays an important and diverse role in regulating both the quantity and the quality of gene expression. Viral RNAs need to successfully navigate around this cellular machinery to initiate and maintain a highly productive infection. Recent work has shown that viruses have developed a variety of strategies to accomplish this, including inherent RNA shields, hijacking host RNA stability factors, incapacitating the host decay machinery and changing the entire landscape of RNA stability in cells using virally encoded nucleases. In addition to maintaining the stability of viral transcripts, these strategies can also contribute to the regulation and complexity of viral gene expression as well as to viral RNA evolution. PMID- 21640426 TI - Neuroglobin and myoglobin in non-small cell lung cancer: expression, regulation and prognosis. AB - Globins are respiratory proteins involved in oxygen metabolism, which is a critical factor in tumor growth and progression. The status of neuroglobin and myoglobin is largely unknown in human malignancies, including lung cancer. The aim of this study was to explore mRNA expression profiles, potential regulatory mechanisms and clinicopathological associations of neuroglobin and myoglobin in non-small cell lung cancer (NSCLC). We screened 208 surgically resected NSCLC specimens and a panel of lung normal and cancer cell lines. The mRNA expression of neuroglobin, myoglobin and hypoxia markers (HIF1alpha and VEGFa) was measured with qRTPCR, while neuroglobin promoter methylation was assessed with Pyrosequencing. Neuroglobin and myoglobin were upregulated in the tumor samples compared to normal tissue (p=1.3*10(-22) and p=1.9*10(-9), respectively). Neuroglobin was more frequently overexpressed in squamous cell carcinomas (SqCCL) than adenocarcinomas. Overexpression of myoglobin was more profound in adenocarcinomas, which correlated with poor survival (p=0.013). Neuroglobin promoter was hypermethylated in 30.8% of NSCLC cases, which correlated with neuroglobin mRNA downregulation. The epigenetic regulation of neuroglobin was confirmed by treating lung cell lines with 5'azadeoxycytidine and/or trichostatin A. Expression of both genes correlated with the expression of HIF1alpha (neuroglobin: p=3.8*10(-5), myoglobin: p=1.1*10(-11)). Myoglobin expression was also associated to that of VEGFa (p=2.1*10(-7)). Hypoxia-dependent upregulation of both globins was validated in vitro. In summary, neuroglobin and myoglobin overexpression in NSCLC is associated with histological subtype, hypoxia and, in case of neuroglobin - epigenetic regulation. Myoglobin expression may have potential significance in the prognostication of lung adenocarcinomas. PMID- 21640427 TI - Urokinase receptor forms in serum from non-small cell lung cancer patients: relation to prognosis. AB - OBJECTIVES: To study the prognostic impact of the different forms of the receptor for urokinase plasminogen activator (uPAR) in serum from 171 non-small cell lung cancer (NSCLC) patients. MATERIALS AND METHODS: Serum sampled preoperatively was available from 171 patients radically resected for NSCLC. Intact uPAR, uPAR(I III), intact and cleaved uPAR, uPAR(I-III)+uPAR(II-III) and the liberated uPAR(I) were measured by time-resolved fluorescence immunoassays (TR-FIAs 1-3). RESULTS: High serum levels of each of the three uPAR forms were associated with short overall survival (OS). In a multivariate survival analysis uPAR(I-III) (hazard ratio (HR)=2.3, 95% confidence interval (CI): 1.2-4.5, p=0.015) and uPAR(I) (hazard ratio (HR)=1.5, 95% CI: 1.0-2.2, p=0.0497) remained significant prognostic parameters independent of stage, histology, age, performance status and therapy. CONCLUSIONS: This retrospective study shows that uPAR(I-III) and uPAR(I) in serum are independent prognostic factors in patients radically operated for NSCLC. Further prospective studies are needed to validate these markers for clinical use. PMID- 21640428 TI - Vacuolar H(+)-ATPases: intra- and intermolecular interactions. AB - V-ATPases in eukaryotes are heteromultimeric, H(+)-transporting proteins. They are localized in a multitude of different membranes and energize many different transport processes. Unique features of V-ATPases are, on the one hand, their ability to regulate enzymatic and ion transporting activity by the reversible dissociation of the catalytic V(1) complex from the membrane bound proton translocating V(0) complex and, on the other hand, their high sensitivity to specific macrolides such as bafilomycin and concanamycin from streptomycetes or archazolid and apicularen from myxomycetes. Both features require distinct intramolecular as well as intermolecular interactions. Here we will summarize our own results together with newer developments in both of these research areas. PMID- 21640429 TI - How do I begin? Sensing extracellular stress to maintain yeast cell wall integrity. AB - The cell wall integrity (CWI) signalling pathway is necessary to remodel the yeast cell wall during normal morphogenesis and in response to cell surface stress. In the Baker's yeast Saccharomyces cerevisiae, a set of five membrane spanning sensors, namely Wsc1, Wsc2, Wsc3, Mid2 and Mtl1, detect perturbations in the cell wall and/or the plasma membrane and activate a downstream signal transduction pathway with a central MAP kinase module. As a consequence, the expression of genes whose products are involved in cell wall structure and remodelling is induced. This review summarises our recent results on sensor structure and function, as well as the advances made regarding sensor mechanics. PMID- 21640430 TI - Virus-specific T cells for therapy--approaches, problems, solutions. AB - Adoptive T cell therapy is the transfer of T cells to a patient in order to combat disease. This procedure is mainly being used but not limited to the treatment of viral infections and malignancies including virus-associated tumors. Depending on the clinical context, the T cell donor may be the same patient or another donor, usually a healthy person. Recent research is centered on the use of antigen-specific T cells, but T cells of uncharacterized specificity can be successfully used in some clinical conditions where target antigens are not known. Depending on underlying scientific hypotheses and preferred technologies, the therapeutic T cells may be anything from monoclonal to highly polyclonal; they may be specific for one epitope, several epitopes from one antigen, or various antigens; they may have been selected during the preparation process for their specificity, their functional capacity, their survival and proliferation in vitro, or the expression of surface markers associated with desirable functional properties. In this minireview, we give a brief overview on selected approaches, problems and solutions in adoptive T cell therapy. We focus on an area where T cell therapy has been particularly successful but is still calling for improvement: herpesviral disease in patients after transplantation. PMID- 21640431 TI - Changes and induction of aminopeptidase activities in response to pathogen infection during germination of pigeonpea (Cajanas cajan) seeds. AB - Aminopeptidases play important role in the mobilization of storage proteins at the cotyledon during seed germination. It is often referred as inducible component of defense against herbivore attack. However the role of aminopeptidase in response to pathogen attack in germinating seeds is remained to be unknown. An attempt was made to analyze change in the aminopeptidase (EC 3.4.11.1) activity during germination of pigeonpea (Cajanus cajan L.) seeds by infecting the seeds with fungi. Two aminopeptidase activity bands (AP1 and AP2) were detected in control as well as infected pigeonpea seeds. During latter stages of germination in control seeds, AP1 activity was replaced by AP2 activity. However AP1 activity was significantly induced in germinating seeds infected with Fusarium oxysporum f.sp. ciceri and Aspergillus niger var. niger. The estimated molecular weights of AP1 and AP2 were ~97 and 42.8kDa respectively. The induced enzyme was purified up to 30 fold by gel filtration chromatography. The purified enzyme was preferentially cleaved leucine p-nitroanilide than alanine p-nitroanilide. The enzyme was strongly inhibited by bestatin and 1,10-phenanthroline. Almost 50% of enzyme activity was inhibited by ethylene diamine tetra acetate. The purified enzyme showed broad pH optima ranging from pH 6.0 to 9.0 and optimum at pH 8.5. The induction of aminopeptidase activity during pigeonpea seed germination and in response to pathogen attack indicates significant involvement of these enzymes in primary as well as secondary metabolism of the seeds. These findings could be helpful to further dissect defensive role of aminopeptidases in seed germination which is an important event in plant's life. PMID- 21640432 TI - Risk factors associated with the conversion of meticillin-resistant Staphylococcus aureus colonisation to healthcare-associated infection. AB - The objective of the study was to identify risk factors for healthcare-associated meticillin-resistant Staphylococcus aureus (HA-MRSA) infections in patients with MRSA colonisation over an extended time period. This was a case-control study conducted at a community teaching hospital. Patients included 41 cases and 82 controls, aged >=18 years, who were nares colonisation culture positive for MRSA and either did or did not develop an HA-MRSA infection within 60 days after index colonisation, respectively. Potential risk factors evaluated included: patient demographics, comorbid conditions, medication use, presence of invasive devices, presence of wounds or other infections, nutritional status, number of hospitalisations and time to infection development. In the univariate analysis, the presence of peripheral vascular disease, three or more comorbidities, a central venous catheter, a Foley catheter, or two or more hospitalisations were significantly associated with increased risk for HA-MRSA infection. Multivariate analysis yielded a model that included presence of a central venous catheter (OR: 8.00; 95% CI: 3.13-20.4) or two or more hospitalisations (OR: 3.37; 95% CI: 1.37 8.26) as independent risk factors for MRSA infection in those with MRSA colonisation. In conclusion, risk factors independently associated with the conversion of MRSA colonisation to HA-MRSA infection include the presence of a central venous catheter or two or more hospitalisations. Strategies involving risk factor minimisation may be helpful in reducing HA-MRSA infections in this patient population. PMID- 21640433 TI - Preventing infection in general surgery: improvements through education of surgeons by surgeons. AB - Surgical patients are at particular risk of healthcare-associated infection (HCAI) due to the presence of a surgical site leading to surgical site infection (SSI), and because of the need for intravascular access resulting in catheter related bloodstream infection (CRBSI). A two-year initiative commenced with an initial audit of surgical practice; this was used to inform the development of a targeted educational initiative by surgeons specifically for surgical trainees. Parameters assessed during the initial audit and a further audit after the educational initiative were related to intra- and postoperative aspects of the prevention of SSIs, as well as care of peripheral venous catheters (PVCs) in surgical patients. The proportion of prophylactic antibiotics administered prior to incision across 360 operations increased from 30.0% to 59.1% (P<0.001). Surgical site dressings were observed in 234 patients, and a significant decrease was found in the percentage of dressings that were tampered with during the initial 48h after surgery (16.5% vs 6.2%, P=0.030). In total, 574 PVCs were assessed over the two-year period. Improvements were found in the proportion of unnecessary PVCs in situ (37.9% vs 24.4%, P<0.001), PVCs in situ for >72h (10.6% vs 3.1%, P<0.001) and PVCs covered with clean and intact dressings (87.3% vs 97.6%, P<0.001). Significant improvements in surgical practice were established for the prevention of SSI and CRBSI through a focused educational programme developed by and for surgeons. Potentially, other specific measures may also be warranted to achieve further improvements in infection prevention in surgical practice. PMID- 21640434 TI - Expressed breast milk on a neonatal unit: a hazard analysis and critical control points approach. AB - With the increasing use of human milk and growing evidence of the benefits of mother's milk for preterm and ill newborns, guidelines to ensure its quality and safety are an important part of daily practice in neonatal intensive care units. Operating procedures based on hazard analysis and critical control points can standardize the handling of mother's expressed milk, thereby improving nutrition and minimizing the risk of breast milk-induced infection in susceptible newborns. Because breast milk is not sterile, microorganisms can multiply when the milk is not handled properly. Additional exogenous contamination should be prevented. Strict hygiene and careful temperature and time control are important during the expression, collection, transport, storage, and feeding of maternal milk. In contrast to formula milk, no legal standards exist for the use of expressed maternal milk. The need for additional measures, such as bacteriological screening or heat treatment, remains unresolved. PMID- 21640435 TI - [Efficacy and safety of non-permanent transvenous pacemaker implantation in an intensive care unit]. AB - OBJECTIVE: To analyze the clinical indications for use, morbidity and mortality associated with a non-permanent transvenous pacemaker. DESIGN: Prospective and observational study. SETTING: Cardiac intensive care unit. METHOD: One hundred and eighty-two patients with non-permanent pacemakers implanted consecutively over a period of four years. DATA COLLECTED: Main variables of interest were demographic data, clinical indications, access route, length of stay and complications. RESULTS: A total of 63% were men, with a median age of 78 +/- 9.5 years and with symptomatic third-degree atrioventricular block in 76.9% of the cases. Femoral vein access was preferred in 92.3% of the cases. Complications appeared in 40.11% of the patients, the most frequent being hematoma at the site of vascular access (13.19%). Restlessness was associated to the need for repositioning the pacemaker due to a shift in the electrode (p=0.059) and to hematoma (p=0.07). Subclavian or jugular vein lead insertion (p=0.012; OR=0.16; 95%CI, 0.04-0.66), restlessness during admission to ICU (p=0.006; OR=3.2; 95%CI, 1.4-7.3), and the presence of cardiovascular risk factors (p=0.042; OR=5; 95%CI, 1.06-14.2) were identified by multivariate analysis as being predictors of complications. Length of stay in ICU was significantly longer when lead insertion was carried out by specialized staff (p=0.0001), and in the presence of complications (p=0.05). CONCLUSIONS: Predictfurors of complications were restlessness, cardiovascular risk factors, and insertion through the jugular or subclavian vein. Complications prolonged ICU stay and were not related to the professionals involved. PMID- 21640436 TI - [Therapeutic possibilities of stem cells in the treatment of liver diseases]. AB - Cell therapy and the use of stem cells in the treatment of liver diseases is still in the research phase. Nevertheless, the diversity of stem cells in terms of their origin, characteristics and potential for differentiation provides a wide spectrum of possibilities for the treatment of liver diseases. The present article describes the main types of stem cells and their potential for the treatment of liver diseases, as well as the main therapeutic strategies that are currently being explored for the treatment of these diseases through cell therapy. In addition, the main preclinical and clinical studies suggesting that stem cells could become an effective therapeutic alternative in distinct liver diseases are discussed. PMID- 21640437 TI - [Psitacosis outbreak in the northern region of Malaga, Spain]. PMID- 21640438 TI - [Which Spanish journals are read and how Primary Care doctors keep informed]. AB - OBJECTIVE: To determine which medical journals are read by Primary Care doctors, their language preferences, publication format and the ways of accessing them. DESIGN: Telephone interview. LOCATION: All the autonomous communities of Spain. PARTICIPANTS: A total of 300 Primary Care physicians selected by proportional stratified sampling by autonomous community and type of professional (general practitioners and paediatricians). MEASURES: A structured 18-question survey on reading habits administered by telephone using computer-assisted techniques. RESULTS: Respondents declared they preferably read specialty journals and are inclined to the printed format, although the online version is the option of younger doctors and paediatricians. The general practitioners prefer to read case studies (73%) and paediatricians opt for the reviews (74%). Overall, 48% read 1 to 4 articles per week. The search tools preferred by 70% of respondents were Google and Yahoo, followed by databases (29%). Eighty eight per cent stated an interest in having a free online service to find quality Spanish journals. As regards language, 80% find it more useful to read articles in Spanish than in English, and 70% believe that there are no quality differences between the publications in both languages. CONCLUSIONS: The publications in Spanish are used as the preferred sources of information by a high percentage of Primary Care physicians. These professionals believe that Spanish-language databases make it easier to access these publications, and are useful and necessary for professional practice. PMID- 21640439 TI - Development of a variable-radius pinhole SPECT system with a portable gamma camera. AB - OBJECTIVE: To develop a small-animal SPECT system using a low cost commercial portable gamma camera equipped with a pinhole collimator, a continuous scintillation crystal and a position-sensitive photomultiplier tube. MATERIAL AND METHODS: The gamma camera was attached to a variable radius system, which enabled us to optimize sensitivity and resolution by adjusting the radius of rotation to the size of the object. To investigate the capability of the SPECT system for small animal imaging, the dependence of resolution and calibration parameters on radius was assessed and acquisitions of small phantoms and mice were carried out. RESULTS: Resolution values, ranging from 1.0mm for a radius of 21.4mm and 1.4mm for a radius of 37.2mm were obtained, thereby justifying the interest of a variable radius SPECT system. CONCLUSIONS: The image quality of phantoms and animals were satisfactory, thus confirming the usefulness of the system for small animal SPECT imaging. PMID- 21640440 TI - [Molecular neuroimaging in the study of cognitive impairment: contribution of the cerebral blood flow SPECT with 99mTc-HMPAO and 18F-FDG PET/CT scan]. AB - OBJECTIVE: The aim of this study was to analyze cerebral perfusion and glucose metabolism in patients with cognitive impairment using cerebral blood flow (99m)Tc-HMPAO SPECT and (18)F-FDG PET/CT scans. MATERIAL AND METHODS: Twenty-two patients with cognitive impairment were included: 4 subjective memory complaints (SMC), 8 amnestic mild cognitive impairment (MCI), 5 prodromic Alzheimer's disease (AD) and 5 AD. In each clinical group, (99m)Tc-HMPAO SPECT and (18)F-FDG PET/CT scans were performed. RESULTS: (99m)Tc-HMPAO SPECT showed regions of cerebral hypoperfusion in 15 patients and was normal in 7 of the 22 patients. (18)F-FDG PET/CT scan showed cerebral regional hypometabolism in 19 patients and was normal in the other 3 patients. The distribution of abnormalities on (99m)Tc HMPAO SPECT and (18)F-FDG PET/CT scans was similar in 9 patients (2 SMC, 2 amnestic MCI, 2 prodromic AD, and 3 AD). In 6 patients (1 amnestic MCI, 2 prodromic AD, and 3 AD), FDG hypometabolism was more extensive than the cerebral hypoperfusion. Four patients (1SMC, 3 amnestic MCI) had an abnormal (18)F-FDG PET/CT scan and normal (99m)Tc-HMPAO SPECT. There were 3 patients (1 SMC 2, amnestic MCI) with normal (99m)Tc-HMPAO SPECT and (18)F-FDG PET/CT scans. CONCLUSION: (99m)Tc-HMPAO SPECT and (18)F-FDG PET/CT scans showed cerebral hypoperfusion and hypometabolism in patients with cognitive impairment, even in patients with clinical diagnosis of SMC. In patients with a normal cerebral blood flow SPECT, brain glucose cerebral hypometabolism can be detected. In some patients, the extension of FDG hypometabolism is more pronounced than that corresponding to the hypoperfusion area. PMID- 21640441 TI - Clinical relevance of incidental finding of focal uptakes in the colon during 18F FDG PET/CT studies in oncology patients without known colorectal carcinoma and evaluation of the impact on management. AB - AIMS: To assess the significance and the impact of focal FDG uptake in the colon in oncology patients without known colorectal carcinoma. MATERIALS AND METHODS: A retrospective study was undertaken on 2,220 (18)F-FDG PET/CT studies carried out consecutively in the Nuclear Medicine Department in our hospital from 2 December 2008 to 31 May 2010. Inclusion criteria were patients with abnormal (18)F-FDG uptake in colorectal area that could not be explained (or not previously known) by their clinical histories. Patients previously diagnosed with colorectal carcinoma were excluded. A total of 86 patients (57 male, average age 63.4, range 46-85) were finally included. Colonoscopy with biopsy was established as a reference test. The impact of these findings on the diagnostic-therapeutic management in these patients was evaluated. RESULTS: A colonoscopy was performed in 54 of the 86 patients, this examination not having been done up-to-date in the remaining 32 patients. Biopsy was obtained in 43 lesions of the 54 patient in whom a colonoscopy was performed. Colon disease was detected in 49 of these 54 patients, obtaining 54 FDG incidental foci which corresponded to 10 previously unsuspected primary colorectal carcinoma, 3 metastases, 27 adenomatous polyps with different degrees of dysplasia and 14 inflammatory processes. In the remaining 5 patients, the colonoscopy was normal. PET/CT modified the diagnostic and treatment management in most of the patients (49/54, that is 91%). CONCLUSIONS: These results confirm the need to determine the cause of abnormal (18)F-FDG colorectal uptakes in the PET/CT studies by using colonoscopy and biopsy. This approach allows for the detection and early treatment of malignant and premalignant lesions. PMID- 21640442 TI - [REM sleep behaviour disorder and "pure" autonomic failure. Presentation of two cases]. PMID- 21640443 TI - [Cost-effectiveness analysis of pneumococcal vaccination in Spain]. AB - OBJECTIVE: To perform a cost-effectiveness analysis of pediatric pneumococcal vaccination in Spain. METHODS: A deterministic population-based model in the form of a decision-tree with a 1-year time horizon was used to estimate the impact of vaccination with Synflorix(r) or Prevenar13(r) in children aged less than 2 years in Spain from the perspective of the National Health System. Epidemiological data were obtained from the hospital discharge minimum data set (MDS) and the literature. Data on costs were obtained from national health costs databases. The main outcomes measured were the number of cases avoided of invasive pneumococcal disease, acute otitis media (AOM) and myringotomies, and hospital admissions for pneumonia. All costs were expressed in 2010 euros. Effectiveness was measured as the number of quality-adjusted life years (QALYs) gained. RESULTS: The efficacy of Synflorix(r) in preventing episodes of AOM caused by non-typeable Haemophilus influenzae could lead to additional prevention of 41,513 episodes of AOM, 36,324 antibiotic prescriptions and 382 myringotomy procedures and 143 QALYs gained compared with Prevenar13(r). The total vaccination cost with Synflorix(r) would result in savings of 22 million euros. CONCLUSIONS: Pneumococcal vaccination with Synflorix(r) in comparison with Prevenar13(r) in children aged less than 2 years old in Spain could achieve a higher number of QALYs and a substantial cost offset. Vaccination with Synflorix(r) would be a dominant strategy in terms of cost-effectiveness. PMID- 21640444 TI - Consensus virtual screening approaches to predict protein ligands. AB - In order to exploit the advantages of receptor-based virtual screening, namely time/cost saving and specificity, it is important to rely on algorithms that predict a high number of active ligands at the top ranks of a small molecule database. Towards that goal consensus methods combining the results of several docking algorithms were developed and compared against the individual algorithms. Furthermore, a recently proposed rescoring method based on drug efficiency indices was evaluated. Among AutoDock Vina 1.0, AutoDock 4.2 and GemDock, AutoDock Vina was the best performing single method in predicting high affinity ligands from a database of known ligands and decoys. The rescoring of predicted binding energies with the water/octanol partition coefficient did not lead to an improvement averaged over ten receptor targets. Various consensus algorithms were investigated and a simple combination of AutoDock and AutoDock Vina results gave the most consistent performance that showed early enrichment of known ligands for all receptor targets investigated. In case a number of ligands is known for a specific target, every method proposed in this study should be evaluated. PMID- 21640445 TI - Synthesis and preliminary in vitro biological evaluation of new carbon-11-labeled celecoxib derivatives as candidate PET tracers for imaging of COX-2 expression in cancer. AB - The enzyme cyclooxygenase-2 (COX-2) is overexpressed in a variety of malignant tumors. This study was designed to develop new radiotracers for imaging of COX-2 in cancer using biomedical imaging technique positron emission tomography (PET). Carbon-11-labeled celecoxib derivatives, [(11)C]4a-c and [(11)C]8a-d, were prepared by O-[(11)C] methylation of their corresponding precursors using [(11)C]CH(3)OTf under basic conditions and isolated by a simplified solid-phase extraction (SPE) method in 52 +/- 2% (n = 5) and 57 +/- 3% (n = 5) radiochemical yields based on [(11)C]CO(2) and decay corrected to end of bombardment (EOB). The overall synthesis time from EOB was 23 min, the radiochemical purity was >99%, and the specific activity at end of synthesis (EOS) was 277.5 +/- 92.5 GBq/MUmol (n = 5). The IC(50) values to block COX-2 for known compounds celecoxib (4d), 4a and 4c were 40, 290 and 8 nM, respectively, and preliminary findings from in vitro biological assay indicated that the synthesized new compounds 4b and 8a-d display similar strong inhibitory effectiveness in the MDA-MB-435 human cancer cell line in comparison with the parent compound 4d. These results encourage further in vivo evaluation of carbon-11-labeled celecoxib derivatives as new potential PET radiotracers for imaging of COX-2 expression in cancer. PMID- 21640446 TI - Lecturers' accounts of facilitating clinical supervision groups within a pre registration mental health nursing curriculum. AB - In 1994 the Department of Health recommended that nurses be introduced to the process of clinical supervision during pre-registration training. Since then a body of literature has emerged, largely focusing on students' experiences of a variety of training initiatives in supervision. There is however a paucity of literature exploring the experiences of nursing lecturers engaged in such initiatives. This paper reports the findings from one part of a three-year prospective longitudinal study examining mental health students' and lecturers' experiences of group clinical supervision undertaken as part of a pre registration course. In this part of the study eight mental health nursing lecturers participated in semi-structured individual interviews in which they discussed their experiences of facilitating student supervision groups. Content analysis of the interview data produced eight major categories: 'attitudes to supervision'; 'perceptions of the student experience'; 'preparation and support'; 'approaches to supervision'; 'the "good" supervisor'; 'the lecturer as supervisor'; 'the structure and process of sessions; and 'the content of supervision'. The findings suggest that the idea of undertaking supervision for students is attractive to lecturers. However, several issues need to be addressed if this type of initiative is to be successful in preparing students for their future role as supervisee. PMID- 21640447 TI - Conceptions of health promotion held by pre-registration student nurses in four schools of nursing in New Zealand. AB - A research project conducted in four nursing schools in New Zealand presents an account of student nurses' conceptions of health promotion. Focus groups of six to ten pre-registration nursing students were conducted in each of the schools of nursing. The findings indicated that the student nurses had difficulty clearly articulating the concept of health education and health promotion. The student nurses focused on individuals. The main strategy they used was the giving of information. This strategy was in keeping with a traditional approach to health promotion. However a number of students did include empowering interpersonal strategies in their health promotion activities. Many of the students demonstrated an awareness of the impact of economic determinants on health but did not have the support or opportunity to address these. None of the students included addressing the political determinants of health in their discussion of the health promoting role of the nurse. PMID- 21640448 TI - Assessment of student nurses in practice: a comparison of theoretical and practical assessment results in England. AB - This study was undertaken in response to concerns raised by Duffy (2003) that assessors of practice were reluctant to fail student nurses in assessments. This generated doubts about the fitness to practice of some registered nurses. An investigation was undertaken into whether quantitative evidence supported the view that pre-registration nurses rarely failed practical assessments. Comparative failure rates from theoretical and practical assessments were requested from all 52 universities in England that offered pre-registration nursing programmes. Responses were received from 27. Findings indicated that a very small proportion of students failed practical assessments; failure rates for theory outstripped practice by a ratio of 5:1. A quarter of universities failed no students in practice. Students were most likely to fail in year one and least likely in year three. This study supports the belief that assessors of practice are reluctant to fail student nurses. It raises a number of questions about the influence that the systems and practices of professional bodies and universities have on practical assessment. However it also indicates that some student nurses have failed practical assessments and that some universities do have systems in place to address this issue. PMID- 21640449 TI - Emotion work in paramedic practice: the implications for nurse educators. AB - Contemporary paramedic practice involves the emergency assessment, management, treatment and transport of the public with minor and major; potentially life threatening conditions (Caroline, 2008). Paramedics are exposed to a myriad of human emotions related to suffering, pain, devastating trauma and death. Managing their own and others emotions appears to be a crucial component of their work, however there is a paucity of evidence relating to this aspect of the role. The movement of preregistration paramedic education into higher education has impacted on nurse educators in some settings, who are undertaking key administrative, managerial, teaching and personal tutoring roles. This is a valuable opportunity for nurse educators to contribute to the education of student paramedics in the crucial area of emotion work as part of interprofessional health care education. This paper examines emotional labour in health care, historical influences on paramedic education, the limited empirical research and the impact of gender, which provide insights into this profession crucial for nurse educators. The implications of emotion work for the educational curriculum and the support of student paramedics are discussed and strategies such as counselling, reflection and personal tutoring are suggested. Mentorship selection and preparation are highlighted and the need for a cultural change in attitude towards emotion work. PMID- 21640450 TI - Head and neck cancer patients' perspective of carer burden. AB - There is a growing appreciation of the important role that carers have in supporting patients following treatment for head and neck cancer. We asked patients about the role fulfilled by their carer(s) and the support they give, and for their thoughts on the burden this placed on the carers. We did a cross sectional survey of 751 patients with head and neck cancer who were alive and disease-free using two questionnaires: one combined study-specific questions about carers with questions from Khafif et al., and the other was the University of Washington Quality of Life questionnaire version 4 (UW-QoL). There were 386 replies. Nearly half (46%, 162/354) had carers who were mainly family members. Patients identified their main roles as providing emotional support (75%), taking them to healthcare appointments (67%), cleaning the home (62%), and shopping for food (59%). Around a third felt that their care was a considerable burden, and a similar proportion felt that it was very hard for their carers. Patients over 65 years of age were the most likely to need a considerable amount of care and support, and those with low socio-emotional UW-QoL composite scores were most likely to need a considerable amount of care and support, to regard the burden on their carers as considerable, and to think that it was very hard for their carers to take care of them. The study emphasises the necessity to take account of the needs of carers. More research is required on the patient-carer relationship and how best to support it. PMID- 21640451 TI - The impact of stochastic resonance electrical stimulation and knee sleeve on impulsive loading and muscle co-contraction during gait in knee osteoarthritis. AB - BACKGROUND: Increased impulsive loading and muscle co-contraction during gait have been observed in individuals with knee osteoarthritis. Proprioceptive deficits in this population may contribute to these effects. Proprioception has been shown to improve with the combination of stochastic resonance electrical stimulation and a knee sleeve in knee osteoarthritis. Our goal was to determine whether stochastic resonance stimulation combined with a knee sleeve would decrease impulsive loading rates and muscle co-contraction during gait in knee osteoarthritis. METHODS: Gait kinetics, kinematics and muscle activity were assessed during walking in subjects with knee osteoarthritis during three different conditions: no stochastic resonance/no sleeve (control), stochastic resonance at 75% threshold/sleeve, and no stochastic resonance/sleeve. Loading rates were calculated from the ground reaction force. Muscle co-contraction was calculated from the ratio of vastus lateralis to lateral hamstring activity. Differences between conditions were assessed using a repeated measures analysis of variance (P<0.05). FINDINGS: The 75% threshold/sleeve and sleeve only conditions resulted in increased knee flexion at contact and reduced loading rates compared to the control condition (P<0.05). However, these measures did not significantly differ between the 75% threshold/sleeve and sleeve only conditions. Muscle co-contraction was found to decrease with the 75% threshold/sleeve condition compared to the other conditions. INTERPRETATION: Increased knee flexion and decreased loading rates may be a result of proprioceptive improvements resulting from the sleeve or sleeve/stimulation combination. The stochastic resonance stimulation did not demonstrate an ability to enhance the effects of the sleeve with the exception of reductions in muscle co-contraction. PMID- 21640452 TI - An update of the mutation profile of Factor 13 A and B genes. AB - Mutational reports over the past two decades have accumulated an immense amount of literature for inherited Factor XIII deficiency. However, the genotype and phenotype correlations for inherited Factor XIII deficiency are complicated. While many studies clearly prove a cause and effect relationship for the reported mutations, others are lacking in this regard. The F13B gene remains an elusive component as far as inherited Factor XIII deficiencies are concerned. Also, an in depth analysis into the heterozygous state of this deficiency is also lacking. In this review we have tried to analyze and present an exhaustive amount of mutational data from the past three decades. The source of our mutational data is our website dedicated to Factor XIII deficiencies (www.F13-database.de) as well as literature search done on the Pubmed (www.ncbi.nlm.nih.gov/pubmed). PMID- 21640453 TI - Functional redundancy and food web functioning in linuron-exposed ecosystems. AB - An extensive data set describing effects of the herbicide linuron on macrophyte dominated microcosms was analysed with a food web model to assess effects on ecosystem functioning. We showed that sensitive phytoplankton and periphyton groups in the diets of heterotrophs were gradually replaced by more tolerant phytoplankton species as linuron concentrations increased. This diet shift- showing redundancy among phytoplankton species--allowed heterotrophs to maintain their functions in the contaminated microcosms. On an ecosystem level, total gross primary production was up to hundred times lower in the treated microcosms but the uptake of dissolved organic carbon by bacteria and mixotrophs was less sensitive. Food web efficiency was not consistently lower in the treated microcosms. We conclude that linuron predominantly affected the macrophytes but did not alter the overall functioning of the surrounding planktonic food web. Therefore, a risk assessment that protects macrophyte growth also protects the functioning of macrophyte-dominated microcosms. PMID- 21640454 TI - Arsenic and Mn levels in Isaza (Gymnogobius isaza) during the mass mortality event in Lake Biwa, Japan. AB - The present study measured the concentrations of 25 elements (Li, Mg, V, Cr, Mn, Co, Ni, Cu, Zn, As, Se, Rb, Sr, Mo, Ag, Cd, In, Sn, Sb, Cs, Ba, Hg, Tl, Pb and Bi) in the whole body of Isaza which is an endemic fish species to Lake Biwa, Japan, and compared the values in the specimens from the mass mortality Isaza (MMI) and normal fresh Isaza (NFI). The mean levels of Mn and total As (T-As) were relatively higher in MMI than in NFI. In the T-As, highly toxic inorganic As was detected in MMI. Moreover we found Mn and As concentrations in surface sediment were extremely high and temporally increased. From all these results, we could infer that the dissolution of Mn and As from surface sediment of Lake Biwa might have been one of the cause for the mass mortality of Isaza. PMID- 21640455 TI - A Qualitative Comparative Analysis of factors associated with trends in narrowing health inequalities in England. AB - This study explores why progress with tackling health inequalities has varied among a group of local authority areas in England that were set targets to narrow important health outcomes compared to national averages. It focuses on premature deaths from cancers and cardiovascular disease (CVD) and whether the local authority gap for these outcomes narrowed. Survey and secondary data were used to create dichotomised conditions describing each area. For cancers, ten conditions were found to be associated with whether or not narrowing occurred: presence/absence of a working culture of individual commitment and champions; spending on cancer programmes; aspirational or comfortable/complacent organisational cultures; deprivation; crime; assessments of strategic partnership working, commissioning and the public health workforce; frequency of progress reviews; and performance rating of the local Primary Care Trust (PCT). For CVD, six conditions were associated with whether or not narrowing occurred: a PCT budget closer or further away from target; assessments of primary care services, smoking cessation services and local leadership; presence/absence of a few major programmes; and population turnover. The method of Qualitative Comparative Analysis was used to find configurations of these conditions with either the narrowing or not narrowing outcomes. Narrowing cancer gaps were associated with three configurations in which individual commitment and champions was a necessary condition, and not narrowing was associated with a group of conditions that had in common a high level of bureaucratic-type work. Narrowing CVD gaps were associated with three configurations in which a high assessment of either primary care or smoking cessation services was a necessary condition, and not narrowing was associated with two configurations that both included an absence of major programmes. The article considers substantive and theoretical arguments for these configurations being causal and as pointing to ways of improving progress with tackling health inequalities. PMID- 21640456 TI - Critical health literacy: a review and critical analysis. AB - Though there has been a considerable expansion of interest in the health literacy concept worldwide, there has also been criticism that this concept has been poorly defined, that it stretches the idea of "literacy" to an indefensible extent and more specifically, that it adds little to the existing concerns and intervention approaches of the better established discipline of health promotion. This paper takes as a starting point the expanded model of health literacy advanced by Nutbeam (2000) and addresses these concerns by interrogating the concept of "critical health literacy" in order to draw conclusions about its utility for advancing the health of individuals and communities. The constituent domains of critical health literacy are identified; namely information appraisal, understanding the social determinants of health, and collective action, and as far as possible each are clearly delineated, with links to related concepts made explicit. The paper concludes that an appreciation of work undertaken in a range of different disciplines, such as media studies, medical sociology, and evidence based medicine can enhance our understanding of the critical health literacy construct and help us understand its usefulness as a social asset which helps individuals towards a critical engagement with health information. There is some evidence that aspects of critical health literacy have indeed been found to be a resource for better health outcomes, but more research is needed in this area, both to develop quantitative and qualitative approaches to evaluating health literacy skills, and to offer convincing evidence that investment in programmes designed to enhance critical health literacy are worthwhile. PMID- 21640457 TI - Psychological strains and youth suicide in rural China. AB - The strain theory of suicide postulates that suicide is usually preceded by psychological strains. A strain can be a consequence of any of four conflicts: differential values, aspiration and reality, relative deprivation, and lack of coping skills for a crisis. This study, with a blend of psychiatric and social predictors of suicide, identified correlates of suicide that are relevant to Chinese culture and tested the strain theory of suicide with Chinese data. We sampled 392 suicides and 416 living controls (both aged 15-34 years) from 16 rural counties in China in 2008 and interviewed two informants for each suicide and each control. We found that marriage and religion/religiosity did not distinguish the suicides from the living controls among Chinese rural young women. Religion/religiosity tended to be stronger for suicides than for controls. Psychological strains in the forms of relative deprivation, unrealized aspiration, and lack of coping skills were significantly associated with suicide, even after accounting for the role of mental illness. The strain theory of suicide forms a challenge to the psychiatric model popular in the West, at least in explaining the Chinese suicide. PMID- 21640458 TI - [Macrolane((r)), a too premature indication in breast augmentation. Focusing on current knowledge of the product]. AB - STUDY PURPOSE: For ten years the market for resorbable fillers has seen a large increase. A CE mark is sufficient for its placing on the market, they do not require AMM because they are not considered drugs. The Macrolane((r)) is a hyaluronic acid NASHA gel-based technology, available on the French market since 2007 as filler used in all areas of the body except face and breasts. It is highly crosslinked, which slows its resorption, leaving in place long-term waste. At end of 2008, Macrolane((r)) received a CE mark for breast augmentation. The aim of this paper is to review current scientific knowledge on the MacrolaneTM and list the many uncertainties regarding its recent breast indication. MATERIALS AND METHODS: We reviewed the PubMed literature and study levels of evidence on Macrolane((r)). All AFFSAPS communication and correspondence with the SOFCPRE on hyaluronic acids and particularly MacrolaneTM were collected. RESULTS: At the moment there is no scientific study of high level of evidence which has studied the effects of Macrolane((r)) on breast parenchyma in terms of carcinogenesis or the disruption of radiological monitoring of the breast. The subglandular method of injection remains complex and uncertain especially about the risk of wrong passage. CONCLUSIONS: The Macrolane((r)) is proposed as an alternative less invasive than breast implants. Nevertheless the lack of scientific data on this product led to its non approval by the Food and Drug Administration in the United States. Currently there remains too much uncertainty on this filler for reasonable use plebiscite. It would therefore be preferable to return to the manufacturer the burden of proof of Macrolane((r)) safety and security and limit its use in clinical trials yet. PMID- 21640459 TI - [Integration between cardiology and primary care: impact on clinical practice]. AB - INTRODUCTION AND OBJECTIVES: To assess the impact of a program integrating cardiology and primary care in clinical practice, compared with usual care. The integrated care consists of a hospital cardiologist in each primary care clinic, shared clinical history, joint practice guidelines, consultation sessions, and other coordinating tools. METHODS: Observational, cross-sectional study of 2 series of chronic outpatients: conventional and integrated care. We analyzed patient distribution and the impact on good clinical practice indicators in patients with ischemic heart disease, heart failure and atrial fibrillation, along with primary care practitioner satisfaction and use of resources. RESULTS: We included 3194 patients (1572 usual care, 1622 integrated care). Integrated care changed the patient distribution, allowing the cardiologist to focus on serious pathologies while cardiovascular risk factors and stable patients were monitored in primary care. In ischemic heart disease, improvement was observed in cholesterol management and blood pressure control; optimal medical treatment was more frequently prescribed and ventricular function evaluated more often. In heart failure, beta-blockers treatment increased and functional class was assessed more often. In atrial fibrillation, an increase in anticoagulation prescription and echocardiography evaluation was observed. Satisfaction parameters improved with integrated care. The use of resources was not increased. CONCLUSIONS: Using our integration model, follow-up and chronic treatment of patients with ischemic heart disease, heart failure, and atrial fibrillation were improved. Monitoring of chronic patients was redistributed between primary care and cardiology, and family physicians' satisfaction levels improved. There was no increase in use of resources. Full English text available from: www.revespcardiol.org. PMID- 21640460 TI - [Factors associated with uncontrolled hypertension in patients with and without cardiovascular disease]. AB - INTRODUCTION AND OBJECTIVES: Hypertension is one of the most prevalent and poorly controlled risk factors, especially in patients with established cardiovascular disease (CVD). The aim of this study was to describe the rate of blood pressure (BP) control and related risk factors. METHODS: Multicenter, cross-sectional and observational registry of patients with hypertension recruited from cardiology and primary care outpatient clinics. Controlled BP defined as <140/90 mmHg. RESULTS: 55.4% of the 10 743 patients included had controlled BP and these had a slightly higher mean age. Patients with uncontrolled BP were more frequently male, with a higher prevalence of active smokers, obese patients, and patients with diabetes. The rate of controlled BP was similar in patients with or without CVD. Patients with uncontrolled BP had higher levels of blood glucose, total cholesterol, low density lipoproteins and uric acid. Patients with uncontrolled BP were receiving a slightly higher mean number of antihypertensive drugs compared to patients with controlled BP. Patients with CVD were more frequently receiving a renin-angiotensin-aldosterone axis inhibitor: 83.5% vs. 73.2% (P<.01). Multivariate analysis identified obesity and current smoking as independently associated with uncontrolled BP, both in patients with or without CVD, as well as relevant differences between the two groups on other factors. CONCLUSIONS: Regardless of the presence of CVD, 55% of hypertensive patients had controlled BP. Lifestyle and diet, especially smoking and obesity, are independently associated with lack of BP control. Full English text available from: www.revespcardiol.org. PMID- 21640461 TI - [Prevalence of metabolic syndrome and its components in patients with acute coronary syndrome]. AB - INTRODUCTION AND OBJECTIVES: A large proportion of patients with coronary disease have metabolic syndrome, although the frequency and association of its different components are not well understood. The aim of this study was to determine the prevalence of metabolic syndrome and the combination of its components in a Spanish cohort of patients with acute coronary syndrome. METHODS: Clinical histories of 574 inpatients with acute coronary syndrome in 6 tertiary hospitals were reviewed and the presence of metabolic syndrome and its components determined by applying Adult Treatment Panel III criteria. In a second step, the components of the metabolic syndrome were analyzed, excluding those patients with diabetes mellitus. RESULTS: The metabolic syndrome was present in 50.9% of patients and was more frequent in women than in men (66.3% vs. 47.3%; P<.001). The most prevalent component was carbohydrate metabolism disorder (85.3%), followed by low high-density lipoprotein cholesterol (HDLc) levels (80.5%). In nondiabetic patients, 34.6% had metabolic syndrome and the most prevalent component was low HDLc levels (86%), followed by high blood pressure and hypertriglyceridemia and, in fourth place, impaired fasting serum glucose levels. CONCLUSIONS: The metabolic syndrome has a high prevalence in patients with an acute coronary syndrome, especially in women. The most frequent components are hyperglycemia and low HDLc levels. After excluding diabetic patients, the most prevalent diagnostic criterion of metabolic syndrome was low HDLc levels. Full English text available from: www.revespcardiol.org. PMID- 21640462 TI - [Hematologic diseases: from within the heart]. AB - The fundamental relationship between blood disorders and the cardiovascular system originates within multiple points of interface, ranging from the heart and its structural constituents to include heart chambers, valves, coronary arteries, coronary veins, and the cerebrovascular and peripheral vasculature. While the cellular components of circulating blood derive their primary origin from multipotent progenitor cells, plasma-based components, which include coagulation proteins, are mostly born of hepatic synthesis and endothelial cells. Here we provide a focused overview of nononcological blood disorders and their potential impact on the arterial circulatory system as common phenotypes, including myocardial infarction, ischemic stroke and peripheral arterial occlusive events. Venous thromboembolism is employed in our discussion as a clinical template. We also provide practical steps and guidance for diagnostic testing and management in routine clinical practice. Full English text available from: www.revespcardiol.org. PMID- 21640463 TI - Resolution of IgE-mediated fish allergy. PMID- 21640465 TI - Comparison of the diagnostic efficacy of the two rapid bedside immunoassays and combined clinical conventional diagnosis in prelabour rupture of membranes. AB - OBJECTIVE: Diagnosis of prelabour rupture of membranes (PROM) may be challenging. Conventional diagnostic methods such as speculum examination, ferning pattern, and pH detection are not satisfactorily accurate. Two relatively new, commercially available rapid bedside immunoassay strip tests, for placental alpha microglobulin-1 (PAMG-1) and insulin-like growth factor binding protein-1 (ILGFBP 1), are alleged to be more accurate. We compared the diagnostic efficacy of PAMG 1 and ILGFBP-1 immunoassay tests and combined conventional clinical diagnostic methods in PROM. STUDY DESIGN: 167 pregnant women with signs and/or symptoms of PROM were prospectively evaluated with a combination of conventional clinical tests including speculum examination, nitrazine, ferning pattern, and pooling, in addition to PAMG-1 and ILGFBP-1 immunoassays. Differences in sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy of the diagnostic methods were compared. RESULTS: Although the PAMG-1 assay was more accurate, the PAMG-1 immunoassay, ILGFBP-1 immunoassay, and combined conventional clinical diagnosis were, in fact, all highly accurate with no significant difference in sensitivity, specificity, PPV, NPV, or accuracy. CONCLUSION: Both rapid bedside strip tests may be used in clinical practice with similar efficacy in diagnosing PROM, particularly as a backup when diagnosis is still in doubt following a combination of conventional diagnostic methods. PMID- 21640464 TI - Duration of passive and active phases of the second stage of labour and risk of severe postpartum haemorrhage in low-risk nulliparous women. AB - OBJECTIVE: To assess the risk of severe postpartum haemorrhage (PPH) according to the durations of the passive and active phases of the second stage of labour. STUDY DESIGN: Secondary analysis from the PREMODA prospective observational study in 138 French maternity units; 3330 low-risk nulliparous women with vaginal deliveries of cephalic singletons were included. Prospective analysis of the recorded durations of the active first stage of labour and the passive and active phases of the second stage of labour was undertaken, and their association with severe PPH, defined by estimated blood loss >1000 ml or blood transfusion. Factors associated with severe PPH were analysed by uni and multivariate analyses with logistic regression models. RESULTS: The frequency of severe PPH was 2.1% (n=69). In the univariate analysis, the frequency of severe PPH increased with the duration of the active second stage but not the passive second stage: 1.2% for active second stage <10 min, 1.6% for 10-19 min, 2.1% for 20-29 min, 2.6% for 30-39 min, 4.5% for 40-49 min and 14.3% for >= 50 min (p<0.001). After adjustment for confounding factors, the risk of severe PPH was found to be statistically significant when the active first stage exceeded 6h [adjusted odds ratio (OR) 2.5, 95% confidence interval (CI) 1.0-6.1)] and when the active second stage exceeded 40 min (40-49 min: adjusted OR 3.5, 95% CI 1.0-12.3; >= 50 min: adjusted OR 10.6, 95% CI 2.8-40.3; reference: <10 min). The duration of the active second stage was not associated with other maternal or neonatal complications. CONCLUSIONS: A prolonged active, but not passive, second stage of labour is associated with the risk of severe PPH in nulliparas. The optimal duration of these phases remains to be defined. PMID- 21640466 TI - Effect of the bone marrow derived-mononuclear stem cells transplantation in the growth, VEGF-R and TNF-alpha expression of endometrial implants in Wistar rats. AB - OBJECTIVE: To study the effect of bone marrow derived-mononuclear stem cells transplantation in the growth, VEGF-R and TNF-alpha expression of surgically induced endometriosis in an experimental model. STUDY DESIGN: This is an experimental study conducted in the Center for Health and Biological Sciences at the Pontifical Catholic University of Parana, Brazil. Endometriotic implants were surgically induced in 120 female Wistar rats. The animals with viable endometrial implant (larger than 25 mm(2)) were randomically divided into 3 groups to receive an intraperitoneal injection of 0.2 cc of saline solution (C group; n=30), a subcutaneous injection of 1mg/kg of leuprolide (L group; n=34), or an intraperitoneal injection of 5*10(6) bone marrow derived-mononuclear stem cells (SC group; n=36). They were sacrificed after 21 days to assess the implants' size and the tissue expression of vascular endothelial growth factor receptor (VEGF-R) and tumor necrosis factor-alpha (TNF-alpha). RESULTS: Treatment with leuprolide decreased the surface area of the endometriotic implant compared to the SC group and the C group. The absolute reduction in the surface area of the implant was 16.5mm, 0mm, and 0mm (p=0.007), respectively, and the percent reduction was 40.2%, 0%, and 0% (p=0.001). VEGF-R expression in the endometriotic implant decreased after treatment in the L and SC groups compared to the C group (409.6 MUm(2) vs. 465 MUm(2) vs. 920.9 MUm(2), respectively; p=0.021). TNF-alpha expression also reduced in the L and SC groups compared to the C group (585.7 MUm(2) vs. 549.3 MUm(2) vs. 2402.1 MUm(2), respectively; p<0.001). CONCLUSION: Bone marrow derived-mononuclear stem cells transplantation decreased the expression of VEGF-R and TNF-alpha in the endometriotic implant but did not reduce the surface area of the lesion. PMID- 21640467 TI - Essential thrombocythemia and pregnancy. AB - Essential thrombocythaemia (ET) is an acquired myeloproliferative neoplasm, characterised by persistent thrombocytosis and a tendency for either thrombosis or haemorrhage. Among myeloproliferative neoplasms, ET is the most prevalent in young women, which constitute a special group due to their childbearing potential. An increased risk of fetal and maternal complications has been demonstrated in patients with ET. The most common pregnancy-related complication is spontaneous abortion during the first trimester. Recurrent abortion, fetal growth restriction, stillbirth and placental abruption are less frequent. Maternal complications are relatively rare and essentially represented by thromboembolic and bleeding events. Here we summarize the literature describing pregnancy and its outcome in patients with ET and discuss some recommendations for the management of pregnancy. PMID- 21640468 TI - Stress-sensitive tissue regeneration in viscoelastic biomaterials subjected to modulated tensile strain. AB - This research contribution addresses the mechanochemistry of intra-tissue mass transfer for nutrients, oxygen, growth factors, and other essential ingredients that anchorage-dependent cells require for successful proliferation on biocompatible surfaces. The unsteady state reaction-diffusion equation (i.e., modified diffusion equation) is solved according to the von Karman-Pohlhausen integral method of boundary layer analysis when nutrient consumption and tissue regeneration are stimulated by harmonically imposed stress. The mass balance with diffusion and stress-sensitive kinetics represents a rare example where the Damkohler and Deborah numbers appear together in an effort to simulate the development of mass transfer boundary layers in porous viscoelastic biomaterials. The Boltzmann superposition integral is employed to calculate time-dependent strain in terms of the real and imaginary components of dynamic compliance for viscoelastic solids that transmit harmonic excitation to anchorage-dependent cells. Rates of nutrient consumption under stress-free conditions are described by third-order kinetics which include local mass densities of nutrients, oxygen, and attached cells that maintain dynamic equilibrium with active protein sites in the porous matrix. Thinner nutrient mass transfer boundary layers are stabilized at shorter dimensionless diffusion times when the stress-free intra-tissue Damkohler number increases above its initial-condition-sensitive critical value. The critical stress-sensitive intra-tissue Damkohler number, above which it is necessary to consider the effect of harmonic strain on nutrient consumption and tissue regeneration, is proportional to the Deborah number and corresponds to a larger fraction of the stress-free intra-tissue Damkohler number in rigid biomaterials. PMID- 21640469 TI - Glycosaminoglycans promote fibril formation by amyloidogenic immunoglobulin light chains through a transient interaction. AB - Amyloid formation occurs when a precursor protein misfolds and aggregates, forming a fibril nucleus that serves as a template for fibril growth. Glycosaminoglycans are highly charged polymers known to associate with tissue amyloid deposits that have been shown to accelerate amyloidogenesis in vitro. We studied two immunoglobulin light chain variable domains from light chain amyloidosis patients with 90% sequence identity, analyzing their fibril formation kinetics and binding properties with different glycosaminoglycan molecules. We find that the less amyloidogenic of the proteins shows a weak dependence on glycosaminoglycan size and charge, while the more amyloidogenic protein responds only minimally to changes in the glycosaminoglycan. These glycosaminoglycan effects on fibril formation do not depend on a stable interaction between the two species but still show characteristic traits of an interaction-dependent mechanism. We propose that transient, predominantly electrostatic interactions between glycosaminoglycans and the precursor proteins mediate the acceleration of fibril formation in vitro. PMID- 21640470 TI - A review of the impacts of nature based recreation on birds. AB - Nature based recreation such as wildlife viewing, hiking, running, cycling, canoeing, horse riding and dog walking can have negative environmental effects. A review of the recreation ecology literature published in English language academic journals identified 69 papers from 1978 to 2010 that examined the effect of these activities on birds. Sixty-one of the papers (88%) found negative impacts, including changes in bird physiology (all 11 papers), immediate behaviour (37 out of 41 papers), as well as changes in abundance (28 out of 33 papers) and reproductive success (28 out of 33 papers). Previous studies are concentrated in a few countries (United States, England, Argentina and New Zealand), mostly in cool temperate or temperate climatic zones, often in shoreline or wetland habitats, and mostly on insectivore, carnivore and crustaceovore/molluscivore foraging guilds. There is limited research in some regions with both high bird diversity and nature based recreation such as mainland Australia, Central America, Asia, and Africa, and for popular activities such as mountain bike riding and horse riding. It is clear, however, that non motorised nature based recreation has negative impacts on a diversity of birds from a range of habitats in different climatic zones and regions of the world. PMID- 21640471 TI - Scaled-up bioconversion of fish waste to liquid fertilizer using a 5 L ribbon type reactor. AB - A scaled-up conversion process of fish waste to liquid fertilizer was performed in a 5 L ribbon-type reactor. Biodegradation was performed by inoculation of autoclaved fish waste with 5.84 * 10(5) CFU mL(-1) of mixed microorganisms for 96 h. As a result, the pH changed from 6.92 to 5.72, the cell number reached 7.28 * 10(5) CFU mL(-1), and approximately 430 g (28.3%) of fish waste was degraded. Analyses indicated that the 96 h culture of inoculated fish waste possessed comparable fertilizing ability to commercial fertilizers in hydroponic culture with amino acid contents of 6.91 g 100 g(-1). Therefore, the scaled-up production achieved a more satisfactory fish waste degradation rate (3.61 g h(-1)) than the flask-scale production (0.24 g h(-1)). The biodegraded broth of fish waste at room temperature did not undergo putrefaction for 6 months due to the addition of 1% lactate. PMID- 21640472 TI - Environmental and economic assessment of combined biostabilization and landfill for municipal solid waste. AB - Biostabilization can remove considerable amounts of moisture and degradable organic materials from municipal solid waste (MSW), and can therefore be an effective form of pretreatment prior to landfill. The environmental and economic impacts of two combined processes, active stage biostabilization + sanitary landfill (AL), and active and curing stage biostabilization + sanitary landfill (ACL), were compared with sanitary landfill (SL) for MSW with high moisture content. The results indicated that land requirement, leachate generation, and CH(4) emission in the ACL process decreased by 68.6%, 89.1%, and 87.6%, respectively, and the total cost was reduced by 24.1%, compared with SL. This implies that a combined biostabilization and landfill process can be an environmentally friendly and economically feasible alternative to landfill of raw MSW with high moisture content. Sensitivity analysis revealed that treatment capacity and construction costs of biostabilization and the oxidation factor of CH(4) significantly influenced the costs and benefits of the AL and ACL process at an extremely low land price. When the land price was greater than 100 USD m( 2), it became the dominating factor in determining the cost of treatment and disposal, and the total costs of ACL were reduced to less than 40% of those of SL. PMID- 21640473 TI - Epidermal thickness and biomechanical properties of plantar tissues in diabetic foot. AB - Diabetic foot is a common complication for people with diabetes but it is unclear whether the change is initiated from the skin surface or underneath plantar tissues. This study compared the thickness of epidermis and the thickness and stiffness of the total plantar soft tissue among people with diabetes with or without complications. Seventy-two people with diabetes, including 22 people with neuropathies, 16 foot ulcerations, 34 pure diabetics without complications and 40 healthy controls participated in the study. The thickness of the epidermal layer of the plantar skin was examined using high-frequency ultrasonography; the thickness and stiffness of the total plantar soft tissue were measured by using tissue ultrasound palpation system at the big toe, the first, third and fifth metatarsal heads; and the heel pad. Compared with the control group, the average epidermal thickness of plantar skin was reduced by 15% in people with diabetic foot ulceration and 9% in people with neuropathy, but was increased by 6% in pure diabetics. There was an 8% increase in total thickness of plantar soft tissue in the 3 diabetic groups at all testing sites (all p < 0.05), except the first metatarsal head. The stiffness of plantar soft tissue was increased in all diabetic groups at all testing sites compared with the control (all p < 0.05). The epidermal plantar skin becomes thinner and plantar soft tissues stiffen in people with diabetes, particularly in persons who have neuropathy or ulceration, which increases the risk of tissue breakdown and ulceration formation. PMID- 21640474 TI - Laparoscopic ultrasound enhances diagnosis and localization of insulinoma in pancreatic head and neck for laparoscopic surgery with satisfactory postsurgical outcomes. AB - This study explored the value of laparoscopic ultrasonography (LUS) for tumor localization in laparoscopic pancreatic surgery of insulinomas, especially for tumors located at anatomically unfavorable positions. Twenty-eight patients with insulinomas were enrolled in this study between July 2007 and March 2009. Various image examinations were performed preoperatively. An iU22 ultrasound system equipped with a 5.0-9.0 MHz transducer was used for LUS. The tumor localization and postsurgical outcomes were evaluated. Intraoperative LUS precisely localized 33 insulinomas in 26 of 28 patients, whereas the preoperative imaging studies detected 27 of 33 (82%) tumors. No definite tumor in the pancreas and extra pancreatic organs was identified in two patients by both preoperative and intraoperative imaging examinations. Of 33 tumors, 32 (97%) were localized in the pancreas (14 in the head and neck, 18 in the body and tail), whereas one (3%) was found in the duodenal ligament. Successful laparoscopic resection of insulinoma was performed in 21 of 26 patients, including resection of 11 tumors located in the head and neck of the pancreas. Five patients required conversion to open surgery. All insulinomas were benign with a mean size 13.8 mm. Four patients had pancreatic-related complications that spontaneously healed within 3 weeks after surgery. The median hospital stay was 8.5 days. Our study demonstrates that laparoscopic pancreatic resection under the guidance of advanced LUS is not only feasible and safe for tumors located at the body and tail but also for tumors located at the head and neck of the pancreas. PMID- 21640475 TI - The influence of comorbidities and etiologies on the success of extracorporeal shock wave therapy for chronic soft tissue wounds: midterm results. AB - Possible effects of comorbidities and of different wound etiologies on the success of extracorporeal shock wave therapy (ESWT) of chronic soft tissue wounds were investigated. From September 2003 until February 2007, 282 patients, being previously treated unsuccessfully were enrolled. Treatment consisted of ESWT occurring at defined intervals. At each treatment session a wound bed score was recorded, also at initial presentation a detailed patient history and wound etiology. Observed comorbidities were pooled according to the chapters of the ICD 10 system. Two hunderd fifty-eight patients were analyzed (91.49%) and underwent follow-up for a median of 31.8 months. Wound closure occurred in 191 patients (74.03%) by a median of two treatment sessions. No wound reappeared at the same location. A multivariate logistic regression model showed that pooled comorbidities and wound etiologies did not have a significant influence on success. Comorbidities and wound etiologies have surprisingly no significant influence on the success of ESWT. PMID- 21640476 TI - Fracture healing enhancement with low intensity pulsed ultrasound at a critical application angle. AB - Low-intensity pulsed ultrasound (LIPUS) was shown to have dose-dependent enhancement effect on the osteogenic activity of human periosteal cells that played an important role in fracture healing. It was hypothesized that the stimulatory effects of LIPUS on the periosteal cells could be optimized by adjusting the ultrasound delivered at its critical angle to the surface of bone. This increased the transmission of ultrasound waves on periosteum. By using a rat femoral fracture model, the stimulatory effects of LIPUS transmitted at 0 degrees , 22 degrees , 35 degrees and 48 degrees , and the sham-treatment control were investigated. Treatment efficacy was assessed using radiography, micro-computed tomography (micro-CT), histomorphometry and torsional test. The results showed that callus mineralization and bridging, biomechanical properties were significantly enhanced in the 35 degrees group over the control and 0 degrees groups after week 8. LIPUS transmitted at 35 degrees , which could be the critical application angle, showed the best enhancement effects among all the other groups. LIPUS transmitted at a critical application angle may have greater enhancement effects in fracture healing. PMID- 21640477 TI - Usefulness of gray-scale contrast-enhanced ultrasonography (SonoVue(r)) in diagnosing hepatic alveolar echinococcosis. AB - Hepatic alveolar echinococcosis (HAE) is a parasitic infection with an infiltrative growth pattern that has the appearance of a hepatic malignant tumor. Ultrasound (US) has been used for screening of HAE in epidemic areas. However, it has been very difficult to evaluate the clear boundary and microvessel perfusion of the lesions. The aim of this study was to demonstrate the characteristic imaging and clinical significance of HAE lesions by contrast-enhanced ultrasonography (CEUS). Seventeen patients with 19 HAE lesions were examined in sequence with US, color Doppler flow imaging (CDFI) and then CEUS before any treatment. All the data were compared before surgery. Examined by fundamental US, 47.4% of HAE lesions showed irregular hyperechoic substantive areas and 52.6% appeared as having a mixed echotype with irregular anechoic areas in the central portion of the lesions. The CDFI method indicated no blood flow signals inside any of the 19 lesions. By CEUS, all 19 lesions displayed circular rim enhancement in the peripheral segments and absent enhancement within the central areas of the lesions (a "black hole" effect). As a result, the lesions' margins were clear, irregular and distinct. In general, the sizes of all the HAE lesions observed by CEUS were larger than those obtained by fundamental US. Therefore, CEUS is a simple imaging method and can be a helpful tool for more accurate sizing of HAE lesions and their surrounding invasion range and the proper cut-off margin when radical hepatectomy is needed. PMID- 21640478 TI - Pennation angle dependency in skeletal muscle tissue doppler strain in dynamic contractions. AB - Tissue velocity imaging (TVI) is a Doppler based ultrasound technique that can be used to study regional deformation in skeletal muscle tissue. The aim of this study was to develop a biomechanical model to describe the TVI strain's dependency on the pennation angle. We demonstrate its impact as the subsequent strain measurement error using dynamic elbow contractions from the medial and the lateral part of biceps brachii at two different loadings; 5% and 25% of maximum voluntary contraction (MVC). The estimated pennation angles were on average about 4 degrees in extended position and increased to a maximal of 13 degrees in flexed elbow position. The corresponding relative angular error spread from around 7% up to around 40%. To accurately apply TVI on skeletal muscles, the error due to angle changes should be compensated for. As a suggestion, this could be done according to the presented model. PMID- 21640479 TI - Transcatheter arterial chemoembolization in combination with high-intensity focused ultrasound for unresectable hepatocellular carcinoma: a systematic review and meta-analysis of the chinese literature. AB - To evaluate the therapeutic effects of transcatheter arterial chemoembolization (TACE) in combination with high-intensity focused ultrasound (HIFU) for unresectable hepatocellular carcinoma (UHCC), we performed a systematic review and meta-analysis by searching the database. In nine controlled trials with a total of 736 patients, the effect of TACE plus HIFU (group A) was compared with TACE alone (group B) in treating UHCC. Outcomes mainly included tumor response and survival. Pooled odds ratios (ORs) and 95% confidence intervals (CIs) were calculated for each relationship from raw study data using the fixed-effect model or the random effect model. We found that group A significantly improved the tumor response and 0.5-, 1-, 2-, and 3-year survival of patients. In conclusion, combined therapy was more therapeutically beneficial. However, considering the strength of the evidence, additional randomized controlled trials are needed before combined therapy can be recommended routinely. PMID- 21640480 TI - Serial ultrasound evaluation of intramyocardial strain after reperfused myocardial infarction reveals that remote zone dyssynchrony develops in concert with left ventricular remodeling. AB - This study noninvasively evaluated the development of left ventricular (LV) dyssynchrony following reperfused myocardial infarction (MI) in mice using an ultrasonic speckle-tracking method. Eight C57BL/6J mice were assessed by high resolution echocardiography at baseline and at eight time-points following MI. Images were acquired at 1mm elevational intervals encompassing the entire LV to determine chamber volumes and radial strain. Receiver-operating characteristic (ROC) analysis of regional radial strain was used to segment the three dimensional (3-D) LV into infarct, adjacent and remote zones. This in vivo segmentation was correlated to histologic infarct size (R = 0.89, p < 0.01) in a short-axis, slice-by-slice comparison. The onset of dyssynchrony during LV remodeling was assessed by standard deviation of time to peak radial strain in the infarct, adjacent and remote zones. It was discovered that the form of LV dyssynchrony that develops in the remote zone late after MI does so in concert with the progression of LV remodeling (R = 0.70, p < 0.05). PMID- 21640481 TI - A novel 3-D power Doppler ultrasound approach to the quantification of achilles tendon neovascularity. AB - To investigate the amount of neovascularization in the symptomatic Achilles tendon, we developed a novel three-dimensional (3-D) power Doppler ultrasound scanning system to prospectively examine 10 patients with 12 symptomatic Achilles tendons, as well as 20 asymptomatic Achilles tendons as a control group. The mean volume of neovascularity (VON) in the whole Achilles tendon was calculated during the phase of the cardiac cycle displaying maximum vascularity and also during the phase of the cardiac cycle displaying the minimum. The mean VONs in the control group were found to be 0.41 mm(3) (maximum) and 0.02 mm(3) (minimum), respectively. The mean VONs in patients with painful Achilles tendon were 380 mm(3) (maximum) and 70.3 mm(3) (minimum), respectively. The initial results showed that the 3-D power Doppler ultrasound system could be used to measure the mean VON in controls and in patients with symptomatic Achilles tendinopathy. The results demonstrated a significantly greater VON in the maximum phase compared with the minimum phase as well as in the patient group compared with that in the control group. The system was therefore capable of defining the degree of vascularity in the Achilles tendon. PMID- 21640482 TI - Her2 amplification is significantly more frequent in lymph node metastases from urothelial bladder cancer than in the primary tumours. AB - BACKGROUND: Her2, an alias for the protein of v-erb-b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog (avian), might be an attractive therapeutic target in metastasising bladder cancer. Genotype and phenotype of primary tumours and their metastases may differ. OBJECTIVES: Determine Her2 status in both tumour components to better assess the potential of anti-Her2 therapies. DESIGN, SETTING, AND PARTICIPANTS: Histologic examination revealed lymph node metastases in 150 patients with urothelial bladder cancer clinically staged as N0M0. A tissue microarray was constructed with four tumour samples per patient: two from the primary tumour and two from nodal metastases. Her2 status was determined at the gene level by fluorescence in situ hybridisation (FISH) and at the protein level by immunohistochemistry (IHC). INTERVENTIONS: All patients underwent cystectomy and standardised extended lymphadenectomy. MEASUREMENTS: Overall survival was assessed according to HER2 gene status and protein expression in primary bladder cancers and lymph node metastases. RESULTS AND LIMITATIONS: Her2 amplification was significantly more frequent in lymph node metastases (15.3%) than in matched primary bladder cancers (8.7%; p = 0.003). Her2 amplification in primary tumours was highly preserved in the corresponding metastases as indicated by only one amplified primary tumour without amplification of the metastasis. There was a high concordance in HER2 FISH results between both samples from the primary tumour (kappa = 0.853) and from the metastases (kappa = 0.930). IHC results were less concordant (kappa=0.539 and 0.830). FISH and IHC results were poorly correlated in primary tumours (kappa = 0.566) and metastases (kappa = 0.673). While Her2 amplification in the primary tumour significantly predicted poor outcome (p = 0.044), IHC-based survival prediction was unsuccessful. CONCLUSIONS: Her2 amplification in metastasising bladder cancer is relatively frequent, is homogeneous in each tumour component, and predicts early death. This suggests a high potential for anti-Her2 therapies. For patient selection, FISH might be more accurate than IHC. PMID- 21640483 TI - Impact of venous tumour thrombus consistency (solid vs friable) on cancer specific survival in patients with renal cell carcinoma. AB - BACKGROUND: To our knowledge, the impact of venous tumour thrombus (VTT) consistency in patients affected by renal cell carcinoma (RCC) has never been addressed. OBJECTIVE: To analyse the effect of VTT consistency on cancer-specific survival (CSS). DESIGN, SETTING, AND PARTICIPANTS: We retrospectively analysed 174 consecutive patients with RCC and renal vein or inferior vena cava (IVC) VTT who underwent surgical treatment between 1989 and 2007 at our institute. INTERVENTION: All patients underwent radical nephrectomy and thrombectomy. MEASUREMENTS: Pathologic specimens were reviewed by a single uropathologist. In addition to traditional pathologic features, the morphologic aspect of the tumour thrombus was evaluated to distinguish solid from friable patterns. The prognostic role of thrombus consistency (solid vs friable) on CSS was assessed by means of Cox regression models. RESULTS AND LIMITATIONS: The VTT was solid in 107 patients (61.5%) and friable in 67 patients (38.5%). The presence of a friable VTT increased the risk of having synchronous nodal or distant metastases, higher tumour grade, higher pathologic stage, and simultaneous perinephric fat invasion (all p < 0.05). The median follow-up was 24 mo. The median CSS was 33 mo; the median CSS was 8 mo in patients with a friable VTT and 55 mo in patients with a solid VTT (p < 0.001). On multivariable analyses, the presence of a friable VTT was an independent predictor of CSS (p = 0.02). The power of our conclusion may be somewhat limited by the relatively small study population and the retrospective nature of the study. CONCLUSIONS: In patients with RCC and VTT, the presence of a friable thrombus is an independent predictor of CSS. If our finding is confirmed by further studies, the consistency of the tumour thrombus should be introduced into routine pathologic reports to provide better patient risk stratification. PMID- 21640484 TI - The COMFORT-behavior scale is useful to assess pain and distress in 0- to 3-year old children with Down syndrome. AB - Many pediatric intensive care units use the COMFORT-Behavior scale (COMFORT-B) to assess pain in 0- to 3-year-old children. The objective of this study was to determine whether this scale is also valid for the assessment of pain in 0- to 3 year-old children with Down syndrome. These children often undergo cardiac or intestinal surgery early in life and therefore admission to a pediatric intensive care unit. Seventy-six patients with Down syndrome were included and 466 without Down syndrome. Pain was regularly assessed with the COMFORT-B scale and the pain Numeric Rating Scale (NRS). For either group, confirmatory factor analyses revealed a 1-factor model. Internal consistency between COMFORT-B items was good (Cronbach's alpha=0.84-0.87). Cutoff values for the COMFORT-B set at 17 or higher discriminated between pain (NRS pain of 4 or higher) and no pain (NRS pain below 4) in both groups. We concluded that the COMFORT-B scale is also valid for 0- to 3-year-old children with Down syndrome. This makes it even more useful in the pediatric intensive care unit setting, doing away with the need to apply another instrument for those children younger than 3. PMID- 21640485 TI - Gray wolf (Canis lupus) is a natural definitive host for Neospora caninum. AB - The gray wolf (Canis lupus) was found to be a new natural definitive host for Neospora caninum. Neospora-like oocysts were found microscopically in the feces of three of 73 wolves from Minnesota examined at necropsy. N. caninum-specific DNA was amplified from the oocysts of all three wolves. Oocysts from one wolf were infective for the gamma interferon gene knock out (KO) mice. Viable N. caninum (designated NcWolfUS1) was isolated in cell cultures seeded with tissue homogenate from the infected mouse. Typical thick walled tissue cysts were found in outbred mice inoculated with the parasite from the KO mouse. Tissue stages in mice stained positively with N. caninum-specific polyclonal antibodies. Our observation suggests that wolves may be an important link in the sylvatic cycle of N. caninum. PMID- 21640486 TI - Divergent ghrelin expression patterns in sheep genetically resistant or susceptible to gastrointestinal nematodes. AB - Gastrointestinal nematodes are a major problem for pastoral ruminant production systems. This problem could be reduced by the application of breeding strategies that select for nematode resistant sheep, but no suitable molecular markers are available. Research selection flocks containing lines that are resistant (R) or susceptible (S) to gastrointestinal nematodes provide an excellent resource for discovering selectable markers, and for studying the underlying mechanisms of an effective anti-nematode response. In this study we have used a combination of quantitative real time PCR assays and ELISA to determine if nematode challenge impacts on the expression of the satiety-regulating hormone ghrelin. The expression responses were then compared between the selection flock R and S lines. The results show that the basal levels of ghrelin in plasma were greater than 2-fold higher in nematode naive S line sheep. Three days after a primary nematode challenge divergent ghrelin expression patterns were observed between the selection lines, with levels increasing in R sheep while decreasing in S sheep. After a secondary challenge this trend was repeated, but following a third challenge ghrelin expression levels rose in both R and S sheep, by which time the S animals had acquired an effective immune response to the nematodes, as measured by a significant reduction in faecal egg output. Importantly, this phenomenon was observed in gene expression studies in gut tissues and also in ELISA measurements of ghrelin peptide levels in plasma. A regression analysis showed that ghrelin transcript expression in the gut accounted for >40% of the variation in faecal egg count measured following Haemonchus or Trichostrongylus infection. We therefore hypothesise that the direction of ghrelin expression (up or down) immediately following nematode exposure may play an important role in regulating the differing anti-nematode responses that occur in the R and S lines. Such differences identify ghrelin as a previously unrecognized factor influencing the acquisition of immunity to nematodes. PMID- 21640487 TI - Development and validation of the appearance and performance enhancing drug use schedule. AB - Appearance-and-performance enhancing drug (APED) use is a form of drug use that includes use of a wide range of substances such as anabolic-androgenic steroids (AASs) and associated behaviors including intense exercise and dietary control. To date, there are no reliable or valid measures of the core features of APED use. The present study describes the development and psychometric evaluation of the Appearance and Performance Enhancing Drug Use Schedule (APEDUS) which is a semi-structured interview designed to assess the spectrum of drug use and related features of APED use. Eighty-five current APED using men and women (having used an illicit APED in the past year and planning to use an illicit APED in the future) completed the APEDUS and measures of convergent and divergent validity. Inter-rater agreement, scale reliability, one-week test-retest reliability, convergent and divergent validity, and construct validity were evaluated for each of the APEDUS scales. The APEDUS is a modular interview with 10 sections designed to assess the core drug and non-drug phenomena associated with APED use. All scales and individual items demonstrated high inter-rater agreement and reliability. Individual scales significantly correlated with convergent measures (DSM-IV diagnoses, aggression, impulsivity, eating disorder pathology) and were uncorrelated with a measure of social desirability. APEDUS subscale scores were also accurate measures of AAS dependence. The APEDUS is a reliable and valid measure of APED phenomena and an accurate measure of the core pathology associated with APED use. Issues with assessing APED use are considered and future research is considered. PMID- 21640488 TI - Treatment of frozen shoulder with intra-articular injection of botulinum toxin in shoulder joints. AB - Frozen shoulder, or adhesive capsulitis, is a disease of chronic inflammation and fibrosis involving the shoulder joints. The disease is challenging in clinical practice due to painful treatment course, with disease evolution usually lasting weeks or even months, causing significant discomfort and functional loss. Botulinum toxin (BTX) has been discovered to possess abilities in pain management at various clinical diseases. The actual mechanism is still under investigation, and involves inhibition of neurotransmitter release. In addition, BTX is also found to retard process of fibrosis, which is also a key issue in pathogenesis of frozen shoulder. Therefore, we hypothesize that intra-articular injection of BTX can be an effective alternative for treatment of frozen shoulder. PMID- 21640489 TI - Effect of sex and age on bovine muscle and adipose fatty acid composition and stearoyl-CoA desaturase mRNA expression. AB - The objective of the study was to investigate the effects of sex and slaughter age on the fatty acid (FA) composition and stearoyl-CoA desaturase gene expression in muscle and adipose tissue. Twenty-four Charolais * Simmental crossbred bulls and heifers were raised under similar conditions and slaughtered at 14 or 18 months of age. The total amount of FA in muscle increased markedly in older animals with higher contents of monounsaturated FA (MUFA) in heifers than bulls. The proportions of MUFA and desaturation indices were higher in heifers, whereas polyunsaturated FA were higher in bulls in both muscle and subcutaneous adipose tissue. The results of this study demonstrated sex-dependent differences in the FA composition of muscle and subcutaneous adipose tissue from cattle slaughtered at different ages. The expression of the stearoyl-CoA desaturase gene was higher in the adipose tissue of heifers compared to bulls, and its variation partly contributed to sex- and age-differences in the FA composition of bovine adipose tissue. PMID- 21640490 TI - Accelerated partial breast irradiation is safe and effective using intensity modulated radiation therapy in selected early-stage breast cancer. AB - PURPOSE: To report the feasibility, toxicity, cosmesis, and efficacy of using intensity-modulated radiation therapy (IMRT) with respiratory gating to deliver accelerated partial breast irradiation (APBI) in selected Stage I/II breast cancer after breast-conserving surgery. METHODS AND MATERIALS: Eligible patients with node-negative Stage I/II breast cancer were prospectively enrolled in an institutional review board approved protocol to receive APBI using IMRT after breast-conserving surgery. The target volume was treated at 3.8 Gy/fraction twice daily for 5 days, to a total dose of 38 Gy. RESULTS: Thirty-six patients were enrolled for a median follow-up time of 44.8 months. The median tumor size was 0.98 cm (range, 0.08-3 cm). The median clinical target volume (CTV) treated was 71.4 cc (range, 19-231 cc), with the mean dose to the CTV being 38.96 Gy. Acute toxicities included Grade 1 erythema in 44% of patients and Grade 2 in 6%, Grade 1 hyperpigmentation in 31% of patients and Grade 2 in 3%, and Grade 1 breast/chest wall tenderness in 14% of patients. No Grade 3/4 acute toxicities were observed. Grade 1 and 2 late toxicities as edema, fibrosis, and residual hyperpigmentation occurred in 14% and 11% of patients, respectively; Grade 3 telangiectasis was observed in 3% of patients. The overall cosmetic outcome was considered "excellent" or "good" by 94% of patients and 97% when rated by the physician, respectively. The local control rate was 97%; 1 patient died of a non cancer-related cause. CONCLUSIONS: APBI can be safely and effectively administered using IMRT. In retrospective analysis, IMRT enabled the achievement of normal tissue dose constraints as outlined by Radiation Therapy Oncology Group 04-13/NSABP B-13 while providing excellent conformality for the CTV. Local control and cosmesis have remained excellent at current follow-up, with acceptable rates of acute/late toxicities. Our data suggest that cosmesis is dependent on target volume size. Further prospective multi-institutional trials should be performed to evaluate IMRT to deliver APBI. PMID- 21640491 TI - Clinical outcomes using accelerated partial breast irradiation in patients with invasive lobular carcinoma. AB - PURPOSE: We compared clinical outcomes of women diagnosed with either invasive lobular carcinoma (ILC) or invasive ductal carcinoma (IDC) treated with accelerated partial breast irradiation (APBI). METHODS AND MATERIALS: A total of 16 patients with ILC received APBI as part of their breast-conservation therapy (BCT) and were compared with 410 patients with IDC that received APBI as part of their BCT. Clinical, pathologic, and treatment related variables were analyzed including age, tumor size, hormone receptor status, surgical margins, lymph node status, adjuvant hormonal therapy, adjuvant chemotherapy, and APBI modality. Clinical outcomes including local recurrence (LR), regional recurrence (RR), disease-free survival (DFS), cause-specific survival (CSS), and overall survival (OS) were analyzed. RESULTS: Median follow-up was 3.8 years for the ILC patients and 6.0 years for the IDC patients. ILC patients were more likely to have positive margins (20.0% vs. 3.9%, p = 0.006), larger tumors (14.1 mm vs. 10.9 mm, p = 0.03) and less likely to be node positive (0% vs. 9.5%, p < 0.001) when compared with patients diagnosed with IDC. The 5-year rate of LR was 0% for the ILC cohort and 2.5% for the IDC cohort (p = 0.59). No differences were seen in the rates of RR (0% vs. 0.7%, p = 0.80), distant metastases (0% vs. 3.5%, p = 0.54), DFS (100% vs. 94%, p = 0.43), CSS (100% vs. 97%, p = 0.59), or OS (92% vs. 89%, p = 0.88) between the ILC and IDC patients, respectively. Additionally, when node-positive patients were excluded from the IDC cohort, no differences in the rates of LR (0% vs. 2.2%, p = 0.62), RR (0% vs. 0%), DFS (100% vs. 95%, p = 0.46), CSS (100% vs. 98%, p = 0.63), or OS (92% vs. 89%, p = 0.91) were noted between the ILC and IDC patients. CONCLUSION: Women with ILC had excellent clinical outcomes after APBI. No difference in local control was seen between patients with invasive lobular versus invasive ductal histology. PMID- 21640492 TI - Role of intra- or periprostatic calcifications in image-guided radiotherapy for prostate cancer. AB - PURPOSE: Image-guided radiotherapy (IGRT) allows more precise localization of the prostate, thus minimizing errors resulting from organ motion and set-up during treatment of prostate cancer. Using megavoltage cone-beam computed tomography (MVCBCT), references such as bones, the prostate itself or implanted fiducial markers can be used as surrogates to correct patient positioning immediately before each treatment fraction. However, the use of fiducials requires an invasive procedure and may increase costs. We aimed to assess whether intra- or periprostatic calcifications (IPC) could be used as natural fiducials. METHODS AND MATERIALS: Data on patients treated with IGRT for prostate cancer with clearly visible IPC and implanted fiducials in both planning CT and MVCBCT images were reviewed. IPC were classified as central when inside the prostate and peripheral when within the planning target volume. Daily deviations in lateral, longitudinal, and vertical directions from baseline positioning using fiducials and using IPC were compared. RESULTS: A total of 287 MVCBCT images were obtained and analyzed from 10 patients. The mean +/- standard deviation daily deviation (mm) in the lateral, longitudinal, and vertical coordinates were 0.55 +/- 3.11, 0.58 +/- 3.45, and -0.54 +/- 4.03, respectively, for fiducials, and 0.72 +/- 3.22, 0.63 +/- 3.58, and -0.69 +/- 4.26, for IPC. The p values for comparisons (fiducials vs. IPC) were 0.003, 0.653, and 0.078 for lateral, longitudinal, and vertical coordinates, respectively. When cases with central IPC were analyzed (n = 7), no significant difference was found in such comparisons. Central IPC and fiducials exhibited a similar pattern of displacement during treatment, with equal values for daily displacements in the three directions for more than 90% of measurements. CONCLUSIONS: Our data suggest that centrally located IPC may be used as natural fiducials for treatment positioning during IGRT for prostate cancer, with potential reductions in the risks and costs associated with fiducial implantation. PMID- 21640493 TI - Standard-fractionated radiotherapy for optic nerve sheath meningioma: visual outcome is predicted by mean eye dose. AB - PURPOSE: Radiotherapy has shown its efficacy in controlling optic nerve sheath meningiomas (ONSM) tumor growth while allowing visual acuity to improve or stabilize. However, radiation-induced toxicity may ultimately jeopardize the functional benefit. The purpose of this study was to identify predictive factors of poor visual outcome in patients receiving radiotherapy for ONSM. METHODS AND MATERIALS: We conducted an extensive analysis of 10 patients with ONSM with regard to clinical, radiologic, and dosimetric aspects. All patients were treated with conformal radiotherapy and subsequently underwent biannual neuroophthalmologic and imaging assessments. Pretreatment and posttreatment values of visual acuity and visual field were compared with Wilcoxon's signed rank test. RESULTS: Visual acuity values significantly improved after radiotherapy. After a median follow-up time of 51 months, 6 patients had improved visual acuity, 4 patients had improved visual field, 1 patient was in stable condition, and 1 patient had deteriorated visual acuity and visual field. Tumor control rate was 100% at magnetic resonance imaging assessment. Visual acuity deterioration after radiotherapy was related to radiation-induced retinopathy in 2 patients and radiation-induced mature cataract in 1 patient. Study of radiotherapy parameters showed that the mean eye dose was significantly higher in those 3 patients who had deteriorated vision. CONCLUSIONS: Our study confirms that radiotherapy is efficient in treating ONSM. Long-term visual outcome may be compromised by radiation-induced side effects. Mean eye dose has to be considered as a limiting constraint in treatment planning. PMID- 21640494 TI - 2009 Canadian radiation oncology resident survey. AB - PURPOSE: Statistics from the Canadian post-MD education registry show that numbers of Canadian radiation oncology (RO) trainees have risen from 62 in 1999 to approximately 150 per year between 2003 and 2009, contributing to the current perceived downturn in employment opportunities for radiation oncologists in Canada. When last surveyed in 2003, Canadian RO residents identified job availability as their main concern. Our objective was to survey current Canadian RO residents on their training and career plans. METHODS AND MATERIALS: Trainees from the 13 Canadian residency programs using the national matching service were sought. Potential respondents were identified through individual program directors or chief resident and were e-mailed a secure link to an online survey. Descriptive statistics were used to report responses. RESULTS: The eligible response rate was 53% (83/156). Similar to the 2003 survey, respondents generally expressed high satisfaction with their programs and specialty. The most frequently expressed perceived weakness in their training differed from 2003, with 46.5% of current respondents feeling unprepared to enter the job market. 72% plan on pursuing a postresidency fellowship. Most respondents intend to practice in Canada. Fewer than 20% of respondents believe that there is a strong demand for radiation oncologists in Canada. CONCLUSIONS: Respondents to the current survey expressed significant satisfaction with their career choice and training program. However, differences exist compared with the 2003 survey, including the current perceived lack of demand for radiation oncologists in Canada. PMID- 21640495 TI - Predictors of severe acute and late toxicities in patients with localized head and-neck cancer treated with radiation therapy. AB - PURPOSE: Radiation therapy (RT) causes acute and late toxicities that affect various organs and functions. In a large cohort of patients treated with RT for localized head and neck cancer (HNC), we prospectively assessed the occurrence of RT-induced acute and late toxicities and identified characteristics that predicted these toxicities. METHODS AND MATERIALS: We conducted a randomized trial among 540 patients treated with RT for localized HNC to assess whether vitamin E supplementation could improve disease outcomes. Adverse effects of RT were assessed using the Radiation Therapy Oncology Group Acute Radiation Morbidity Criteria during RT and one month after RT, and the Radiation Therapy Oncology Group/European Organization for Research and Treatment of Cancer Late Radiation Morbidity Scoring Scheme at six and 12 months after RT. The most severe adverse effect among the organs/tissues was selected as an overall measure of either acute or late toxicity. Grade 3 and 4 toxicities were considered as severe. Stepwise multivariate logistic regression models were used to identify all independent predictors (p < 0.05) of acute or late toxicity and to estimate odds ratios (OR) for severe toxicity with their 95% confidence intervals (CI). RESULTS: Grade 3 or 4 toxicity was observed in 23% and 4% of patients, respectively, for acute and late toxicity. Four independent predictors of severe acute toxicity were identified: sex (female vs. male: OR = 1.72, 95% confidence interval [CI]: 1.06-2.80), Karnofsky Performance Status (OR = 0.67 for a 10-point increment, 95% CI: 0.52-0.88), body mass index (above 25 vs. below: OR = 1.88, 95% CI: 1.22-2.90), TNM stage (Stage II vs. I: OR = 1.91, 95% CI: 1.25-2.92). Two independent predictors were found for severe late toxicity: female sex (OR = 3.96, 95% CI: 1.41-11.08) and weight loss during RT (OR = 1.26 for a 1 kg increment, 95% CI: 1.12-1.41). CONCLUSIONS: Knowledge of these predictors easily collected in a clinical setting could help tailoring therapies to reduce toxicities among patients treated with RT for HNC. PMID- 21640496 TI - Definitive radiotherapy for T1-2 hypopharyngeal cancer: a single-institution experience. AB - PURPOSE: To analyze the outcome in T1-2 hypopharyngeal cancer (HPC) patients treated with definitive radiotherapy (RT). PATIENTS AND METHODS: A total of 103 patients with T1-2 hypopharyngeal squamous cell carcinoma treated with radical RT between March 2000 and June 2008 at our institution were analyzed. Pre-RT neck dissection (ND) was performed in 26 patients with advanced neck disease. Chemotherapy was used concurrently with RT in 14 patients. Sixty patients were associated with synchronous or metachronous malignancies. The median follow-up for surviving patients was 41 months. RESULTS: The 3-year overall and cause specific survival rates were 70% and 79%, respectively. The 3-year local control rates were 87% for T1 and 83% for T2 disease. The ultimate local control rate was 89%, including 7 patients in whom salvage was successful. The ultimate local control rate with laryngeal preservation was 82%. Tumors of the medial wall of the pyriform sinus tended to have lower control rates compared with tumors of the lateral or posterior pharyngeal wall. Among patients with N2b-3 disease, the 3 year regional control rates were 74% for patients with pre-RT ND and 40% for patients without ND. The 3-year locoregional control rates were as follows: Stage I, 100%; Stage II, 84%; Stage III, 67%; Stage IVA, 43%; Stage IVB, 67%. Forty-two patients developed disease recurrence, with 29 (70%) patients developing recurrence within the first year. Of the 103 patients, 6 developed late complications higher than or equal to Grade 3. CONCLUSIONS: Definitive RT accomplished a satisfactory local control rate and contributed to organ preservation. PMID- 21640497 TI - Intensity-modulated radiation therapy in oropharyngeal carcinoma: effect of tumor volume on clinical outcomes. AB - PURPOSE: To analyze the effect of primary gross tumor volume (pGTV) and nodal gross tumor volume (nGTV) on treatment outcomes in patients treated with definitive intensity-modulated radiation therapy (IMRT) for oropharyngeal cancer (OPC). METHODS AND MATERIALS: Between September 1998 and April 2009, a total of 442 patients with squamous cell carcinoma of the oropharynx were treated with IMRT with curative intent at our center. Thirty patients treated postoperatively and 2 additional patients who started treatment more than 6 months after diagnosis were excluded. A total of 340 patients with restorable treatment plans were included in this present study. The majority of the patients underwent concurrent platinum-based chemotherapy. The pGTV and nGTV were calculated using the original clinical treatment plans. Cox proportional hazards models and log rank tests were used to evaluate the correlation between tumor volumes and overall survival (OS), and competing risks analysis tools were used to evaluate the correlation between local failure (LF), regional failure (RF), distant metastatic failure (DMF) vs. tumor volumes with death as a competing risk. RESULTS: Median follow-up among surviving patients was 34 months (range, 5-67). The 2-year cumulative incidence of LF, RF and DF in this cohort of patients was 6.1%, 5.2%, and 12.2%, respectively. The 2-year OS rate was 88.6%. Univariate analysis determined pGTV and T-stage correlated with LF (p < 0.0001 and p = 0.004, respectively), whereas nGTV was not associated with RF. On multivariate analysis, pGTV and N-stage were independent risk factors for overall survival (p = 0.0003 and p = 0.0073, respectively) and distant control (p = 0.0008 and p = 0.002, respectively). CONCLUSIONS: In this cohort of patients with OPC treated with IMRT, pGTV was found to be associated with overall survival, local failure, and distant metastatic failure. PMID- 21640498 TI - Greater postimplant swelling in small-volume prostate glands: implications for dosimetry, treatment planning, and operating room technique. AB - PURPOSE: Postimplant prostatic edema has been implicated in suboptimal permanent implants, and smaller prostates have been reported to have worse dosimetric coverage. In this study we compare the degree of postimplant edema between larger and smaller prostates and examine the effects of prostate size on the dose delivered to 90% of the prostate (D90). METHODS AND MATERIALS: From September 2003 to February 2006, 105 hormone-naive patients underwent permanent prostate brachytherapy with (125)I Rapid Strand (Oncura Inc., Arlington Heights, IL). All patients underwent pelvic magnetic resonance imaging (MRI) within 3 weeks before implant, transrectal ultrasound at the time of implant, and both computed tomography and MRI 2.5 to 3 weeks after implant. Prostates were divided into 5 subgroups based on preimplant MRI volumes: less than 25 mL, 25 to 35 mL, 35 to 45 mL, 45 to 55 mL, and greater than 55 mL. Prostate swelling was assessed by use of preimplant and postimplant MRI volumes. Postimplant dosimetry was determined by MRI and compared between the subgroups. RESULTS: All prostates showed postimplant swelling on MRI when compared with preimplant MRI, with a mean increase of 31% +/ 31% (p < 0.0001). The greatest swelling was noted in small prostates (volume less than 25 mL), with a mean increase of 70% +/- 36%. The degree of swelling in the group with a volume less than 25 mL was significantly larger than the degree of swelling in all other prostate subgroups (p < 0.003). Transrectal ultrasound significantly overestimates the prostate volume when compared with MRI by a mean of 15% +/- 25% (p = 0.0006) and is more pronounced for smaller prostates. Although prostates with volumes less than 25 mL did not have significantly worse D90 compared with larger prostates, they had the largest percent of suboptimal implants by the standard ratio of D90 divided by the prescription dose. CONCLUSIONS: Although small prostates have the greatest postimplant edema, planning ultrasound at the time of implant overestimates the volumes of smaller prostates to a greater degree than larger prostates, which may minimize the effects of edema on postimplant dosimetry. PMID- 21640499 TI - N-terminal pro-B-type natriuretic peptide plasma levels as a potential biomarker for cardiac damage after radiotherapy in patients with left-sided breast cancer. AB - PURPOSE: Adjuvant radiotherapy (RT) after breast-conserving surgery has been associated with increased cardiovascular mortality. Cardiac biomarkers may aid in identifying patients with radiation-mediated cardiac dysfunction. We evaluated the correlation between N-terminal pro-B-type natriuretic peptide (NT-proBNP) and troponin (TnI) and the dose of radiation to the heart in patients with left-sided breast cancer. METHODS AND MATERIALS: NT-proBNP and TnI plasma concentrations were measured in 30 left-sided breast cancer patients (median age, 55.0 years) 5 to 22 months after RT (Group I) and in 30 left-sided breast cancer patients (median age, 57.0 years) before RT as control group (Group II). Dosimetric and geometric parameters of heart and left ventricle were determined in all patients of Group I. Seventeen patients underwent complete two-dimensional echocardiography. RESULTS: NT-proBNP levels were significantly higher (p = 0.03) in Group I (median, 90.0 pg/ml; range, 16.7-333.1 pg/ml) than in Group II (median, 63.2 pg/ml; range, 11.0-172.5 pg/ml). TnI levels remained below the cutoff threshold of 0.07 ng/ml in both groups. In patients with NT-proBNP values above the upper limit of 125 pg/ml, there were significant correlations between plasma levels and V(3 Gy)(%) (p = 0.001), the ratios (p = 0.01), the ratios D(15 cm)(3)/D(50%) (Gy) (p = 0.008) for the heart and correlations between plasma levels and V(2 Gy) (%) (p = 0.002), the ratios (p = 0.03), and the ratios (p = 0.05) for the ventricle. CONCLUSIONS: Patients with left-sided breast cancer show higher values of NT-pro BNP after RT when compared with non-RT-treated matched patients, increasing in correlation with high doses in small volumes of heart and ventricle. The findings of this study show that the most important parameters are not the mean doses but instead the small percentage of organ volumes (heart or ventricle) receiving high dose levels, supporting the notion that the heart behaves as a serial organ. PMID- 21640500 TI - The number of pathologically positive lymph nodes and pathological tumor depth predicts prognosis in patients with poorly differentiated squamous cell carcinoma of the oral cavity. AB - PURPOSE: The objective of this retrospective study was twofold: (1) to investigate prognostic factors for clinical outcomes in patients with poorly differentiated oral cavity squamous cell carcinoma and (2) to identify specific prognostic subgroups that may help to guide treatment decisions. METHODS AND MATERIALS: We examined 102 patients with poorly differentiated oral cavity squamous cell carcinoma. All patients were followed for at least 24 months after surgery or until death. The 5-year rates of local control, neck control, distant metastasis, disease-free, disease-specific, and overall survival served as main outcome measures. RESULTS: The 5-year rates were as follows: local control (79%), neck control (64%), distant metastases (27%), disease-free survival (48%), disease-specific survival (52%), and overall survival (42%). Multivariable analysis showed that the number of pathologically positive nodes (>=4 vs. <=3) was a significant predictor of neck control, distant metastasis, and disease free, disease-specific, and overall survival rates. In addition, the presence of tumor depth of >=11 mm (vs. <11 mm) was a significant predictor of distant metastasis, disease-specific survival, and overall survival rates. The combination of the two predictors (26.5%, 27/102) was independently associated with poorer neck control (p = 0.0319), distant metastasis (p < 0.0001), and disease-free (p < 0.0001), disease-specific (p < 0.0001), and overall survival (p < 0.0001) rates. CONCLUSIONS: In patients with poorly differentiated oral cavity squamous cell carcinoma, the presence of at least 4 pathologically positive lymph nodes and of a pathological tumor depth >=11 mm identifies a subset of subjects with poor clinical outcomes. Patients carrying both risk factors are suitable candidates for the development of novel therapeutic approaches. PMID- 21640501 TI - Three-dimensional conformal radiotherapy in prostate cancer patients: rise in interleukin 6 (IL-6) but not IL-2, IL-4, IL-5, tumor necrosis factor-alpha, MIP-1 alpha, and LIF levels. AB - PURPOSE: To investigate the effect of radiotherapy (RT) on serum levels of interleukin-2 (IL-2), IL-4, IL-5, IL-6, tumor necrosis factor alpha (TNF-alpha), macrophage inflammatory protein-1-alpha (MIP-1-alpha) and leukemia inhibitory factor (LIF) in patients with prostate cancer. METHODS AND MATERIALS: Forty eight patients with prostate cancer received three-dimensional conformal blocking radiation therapy with a linear accelerator. IL-2, IL-4, IL-5, IL-6, TNF-alpha, MIP-1-alpha, and LIF levels were measured by the related immunoassay kit 1 day before the beginning of RT and during RT at days 15 and 30. RESULTS: The mean IL 2 values were elevated before and during the RT in contrast with those of IL-4, IL-5, IL-6, TNF-alpha, MIP-1-alpha, and LIF, which were within the normal range under the same conditions. Regarding markers IL-2, IL-4, IL-5, TNF-alpha, MIP-1 alpha, and LIF, comparisons among the three groups (before treatment and 15 and 30 days during RT) did not show significant differences. Although values were within the normal range, there was a significant rise in IL-6 levels at day 15 of RT (p = 0.0049) and a decline at day 30 to levels that were similar to those observed before RT. CONCLUSIONS: IL-6 appeared to peak after 15 days of RT before returning to pre-RT levels. In contrast, IL-2, IL-4, IL-5, TNF-alpha, MIP-1 alpha, and LIF levels were not sensitive to irradiation. The increased levels of IL-6 following RT without the concurrent elevation of other cytokines involved in the acute phase reaction did not suggest a classical inflammatory response to radiation exposure. Further studies should be designed to elucidate the role of IL-6 levels in patients with prostate cancer treated with RT. PMID- 21640502 TI - The influence of radiation modality and lymph node dissection on survival in early-stage endometrial cancer. AB - BACKGROUND: The appropriate uses of lymph node dissection (LND) and adjuvant radiation therapy (RT) for Stage I endometrial cancer are controversial. We explored the impact of specific RT modalities (whole pelvic RT [WPRT], vaginal brachytherapy [VB]) and LND status on survival. MATERIALS AND METHODS: The Surveillance Epidemiology and End Results dataset was queried for all surgically treated International Federation of Gynecology and Obstetrics (FIGO) Stage I endometrial cancers; subjects were stratified into low, intermediate and high risk cohorts using modifications of Gynecologic Oncology Group (GOG) protocol 99 and PORTEC (Postoperative Radiation Therapy in Endometrial Cancer) trial criteria. Five-year overall survival was estimated, and comparisons were performed via the log-rank test. RESULTS: A total of 56,360 patients were identified: 70.4% low, 26.2% intermediate, and 3.4% high risk. A total of 41.6% underwent LND and 17.6% adjuvant RT. In low-risk disease, LND was associated with higher survival (93.7 LND vs. 92.7% no LND, p < 0.001), whereas RT was not (91.6% RT vs. 92.9% no RT, p = 0.23). In intermediate-risk disease, LND (82.1% LND vs. 76.5% no LND, p < 0.001) and RT (80.6% RT vs. 74.9% no RT, p < 0.001) were associated with higher survival without differences between RT modalities. In high-risk disease, LND (68.8% LND vs. 54.1% no LND, p < 0.001) and RT (66.9% RT vs. 57.2% no RT, p < 0.001) were associated with increased survival; if LND was not performed, VB alone was inferior to WPRT (p = 0.01). CONCLUSION: Both WPRT and VB alone are associated with increased survival in the intermediate-risk group. In the high-risk group, in the absence of LND, only WPRT is associated with increased survival. LND was also associated with increased survival. PMID- 21640503 TI - Survival and quality of life after stereotactic or 3D-conformal radiotherapy for inoperable early-stage lung cancer. AB - PURPOSE: To investigate survival and local recurrence after stereotactic ablative radiotherapy (SABR) or three-dimensional conformal radiotherapy (3D-CRT) administered for early-stage primary lung cancer and to investigate longitudinal changes of health-related quality of life (HRQOL) parameters after either treatment. METHODS AND MATERIALS: Two prospective cohorts of inoperable patients with T1-2N0M0 primary lung tumors were analyzed. Patients received 70 Gy in 35 fractions with 3D-CRT or 60 Gy in three to eight fractions with SABR. Global quality of life (GQOL), physical functioning (PF), and patient-rated dyspnea were assessed using the respective dimensions of European Organization for Research and Treatment of Cancer Core Questionnaire-C30 and LC13. HRQOL was analyzed using multivariate linear mixed-effects modeling, survival and local control (LC) using the Kaplan-Meier method, Cox proportional hazards analysis, and Fine and Gray multivariate competing risk analysis as appropriate. RESULTS: Overall survival (OS) was better after SABR compared with 3D-CRT with a HR of 2.6 (95% confidence interval [CI]: 1.5-4.8; p < 0.01). 3D-CRT conferred a subhazard ratio for LC of 5.0 (95% CI: 1.7-14.7; p < 0.01) compared with SABR. GQOL and PF were stable after SABR (p = 0.21 and p = 0.62, respectively). Dyspnea increased after SABR by 3.2 out of 100 points (95% CI: 1.0-5.3; p < 0.01), which is clinically insignificant. At 1 year, PF decreased by an excess of 8.7 out of 100 points (95% CI: 2.8-14.7; p < 0.01) after 3D-CRT compared with SABR. CONCLUSION: In this nonrandomized comparison of two prospective cohorts of medically inoperable patients with Stage I lung cancer, OS and LC were better after SABR. GQOL, PF, and patient-rated dyspnea were stable after SABR, whereas PF decreased after 3D CRT approaching clinical significance already at 1 year. PMID- 21640504 TI - Outcomes of chemoradiotherapy in cervical cancer--the Western Australian experience. AB - PURPOSE: A retrospective review of patients with Stage IB1-IVA cervical cancer treated with combined chemoradiotherapy in Western Australia was conducted with the aim of assessing outcomes and patterns of recurrence. Multivariate analysis was performed to identify potential prognostic factors. METHODS AND MATERIALS: Patients treated with radical chemoradiotherapy for cervical cancer in Western Australia between June 2005 and November 2008 were analyzed. Treatment consisted of external-beam radiotherapy with concurrent weekly cisplatin (40 mg/m(2)), followed by high-dose-rate brachytherapy. The Kaplan-Meier method was used to determine overall survival and disease-free survival, and Cox regression analysis was used to identify potential prognostic factors. RESULTS: Sixty-nine patients were included in the analysis. All patients completed external-beam radiotherapy; however, only 43.5% of patients completed the planned course of brachytherapy. At a median follow-up of 27 months, 24- and 48-month overall survival were 68.8% and 61.1%, respectively. Disease-free survival at 24 and 48 months was 59.4% and 56.7%, respectively. The 2-year local control rate was 70.1%. Nodal involvement resulted in increased risk of disease recurrence (hazard ratio [HR] 6.26, p = 0.002) and death (HR 5.15, p = 0.013). Pretreatment hemoglobin <120 g/L was a negative prognostic factor for disease recurrence (HR 4.20, p = 0.031) and death (HR 8.19, p = 0.020). Completion of brachytherapy improved overall survival (p = 0.039), with a trend to reducing disease recurrence (p = 0.052). The risk of relapse increased with treatment time over 8 weeks (HR 8.18, p = 0.019), however treatment time did not affect the risk of death (p = 0.245). CONCLUSION: The overall survival outcomes in this group of women with locally advanced cervical carcinomas treated with chemoradiotherapy are comparable to worldwide data. Despite the use of modern treatment protocols, a significant proportion of women developed locoregional recurrences and distant failures. Many poor prognostic factors have been identified as contributors to this, including pelvic nodal involvement, incomplete brachytherapy, pretreatment hemoglobin <120 g/L, and treatment time over 8 weeks. PMID- 21640505 TI - Treatment planning constraints to avoid xerostomia in head-and-neck radiotherapy: an independent test of QUANTEC criteria using a prospectively collected dataset. AB - PURPOSE: The severe reduction of salivary function (xerostomia) is a common complication after radiation therapy for head-and-neck cancer. Consequently, guidelines to ensure adequate function based on parotid gland tolerance dose volume parameters have been suggested by the QUANTEC group and by Ortholan et al. We perform a validation test of these guidelines against a prospectively collected dataset and compared with a previously published dataset. METHODS AND MATERIALS: Whole-mouth stimulated salivary flow data from 66 head-and-neck cancer patients treated with radiotherapy at the British Columbia Cancer Agency (BCCA) were measured, and treatment planning data were abstracted. Flow measurements were collected from 50 patients at 3 months, and 60 patients at 12-month follow up. Previously published data from a second institution, Washington University in St. Louis (WUSTL), were used for comparison. A logistic model was used to describe the incidence of Grade 4 xerostomia as a function of the mean dose of the spared parotid gland. The rate of correctly predicting the lack of xerostomia (negative predictive value [NPV]) was computed for both the QUANTEC constraints and Ortholan et al. recommendation to constrain the total volume of both glands receiving more than 40 Gy to less than 33%. RESULTS: Both datasets showed a rate of xerostomia of less than 20% when the mean dose to the least-irradiated parotid gland is kept to less than 20 Gy. Logistic model parameters for the incidence of xerostomia at 12 months after therapy, based on the least-irradiated gland, were D(50) = 32.4 Gy and and gamma = 0.97. NPVs for QUANTEC guideline were 94% (BCCA data), and 90% (WUSTL data). For Ortholan et al. guideline NPVs were 85% (BCCA) and 86% (WUSTL). CONCLUSION: These data confirm that the QUANTEC guideline effectively avoids xerostomia, and this is somewhat more effective than constraints on the volume receiving more than 40 Gy. PMID- 21640506 TI - The relationship between local recurrence and radiotherapy treatment volume for soft tissue sarcomas treated with external beam radiotherapy and function preservation surgery. AB - PURPOSE: To examine the geometric relationship between local recurrence (LR) and external beam radiotherapy (RT) volumes for soft-tissue sarcoma (STS) patients treated with function-preserving surgery and RT. METHODS AND MATERIALS: Sixty of 768 (7.8%) STS patients treated with combined therapy within our institution from 1990 through 2006 developed an LR. Thirty-two received preoperative RT, 16 postoperative RT, and 12 preoperative RT plus a postoperative boost. Treatment records, RT simulation images, and diagnostic MRI/CT data sets of the original and LR disease were retrospectively compared. For LR location analysis, three RT target volumes were defined according to the International Commission on Radiation Units and Measurements 29 as follows: (1) the gross tumor or operative bed; (2) the treatment volume (TV) extending 5 cm longitudinally beyond the tumor or operative bed unless protected by intact barriers to spread and at least 1-2 cm axially (the TV was enclosed by the isodose curve representing the prescribed target absorbed dose [TAD] and accounted for target/patient setup uncertainty and beam characteristics), and (3) the irradiated volume (IRV) that received at least 50% of the TAD, including the TV. LRs were categorized as developing in field within the TV, marginal (on the edge of the IRV), and out of field (occurring outside of the IRV). RESULTS: Forty-nine tumors relapsed in field (6.4% overall). Nine were out of field (1.1% overall), and 2 were marginal (0.3% overall). CONCLUSIONS: The majority of STS tumors recur in field, indicating that the incidence of LR may be affected more by differences in biologic and molecular characteristics rather than aberrations in RT dose or target volume coverage. In contrast, only two patients relapsed at the IRV boundary, suggesting that the risk of a marginal relapse is low when the TV is appropriately defined. These data support the accurate delivery of optimal RT volumes in the most precise way using advanced technology and image guidance. PMID- 21640507 TI - High occurrence of aberrant lymph node spread on magnetic resonance lymphography in prostate cancer patients with a biochemical recurrence after radical prostatectomy. AB - PURPOSE: To investigate the pattern of lymph node spread in prostate cancer patients with a biochemical recurrence after radical prostatectomy, eligible for salvage radiotherapy; and to determine whether the clinical target volume (CTV) for elective pelvic irradiation in the primary setting can be applied in the salvage setting for patients with (a high risk of) lymph node metastases. METHODS AND MATERIALS: The charts of 47 prostate cancer patients with PSA recurrence after prostatectomy who had positive lymph nodes on magnetic resonance lymphography (MRL) were reviewed. Positive lymph nodes were assigned to a lymph node region according to the guidelines of the Radiation Therapy Oncology Group (RTOG) for delineation of the CTV for pelvic irradiation (RTOG-CTV). We defined four lymph node regions for positive nodes outside this RTOG-CTV: the para aortal, proximal common iliac, pararectal, and paravesical regions. They were referred to as aberrant lymph node regions. For each patient, clinical and pathologic features were recorded, and their association with aberrant lymph drainage was investigated. The distribution of positive lymph nodes was analyzed separately for patients with a prostate-specific antigen (PSA) <1.0 ng/mL. RESULTS: MRL detected positive aberrant lymph nodes in 37 patients (79%). In 20 patients (43%) a positive lymph node was found in the pararectal region. Higher PSA at the time of MRL was associated with the presence of positive lymph nodes in the para-aortic region (2.49 vs. 0.82 ng/mL; p = 0.007) and in the proximal common iliac region (1.95 vs. 0.59 ng/mL; p = 0.009). There were 18 patients with a PSA <1.0 ng/mL. Ten of these patients (61%) had at least one aberrant positive lymph node. CONCLUSION: Seventy-nine percent of the PSA-recurrent patients had at least one aberrant positive lymph node. Application of the standard RTOG-CTV for pelvic irradiation in the salvage setting therefore seems to be inappropriate. PMID- 21640508 TI - Assessment of organ motion in postoperative endometrial and cervical cancer patients treated with intensity-modulated radiation therapy. AB - PURPOSE: Intensity-modulated radiation therapy (IMRT) may be useful to reduce toxicity in gynecologic cancer patients requiring postoperative pelvic irradiation. This study was undertaken to quantify vaginal wall organ motion during the course of postoperative pelvic irradiation using pelvic IMRT. METHODS AND MATERIALS: Twenty-two consecutive patients treated with postoperative pelvic IMRT on helical tomotherapy had fiducial markers placed at the vaginal apex prior to simulation then daily megavoltage computed tomography (CT) scans for positioning. The daily positions of the fiducials were registered and measured in reference to the initial CT scan to quantify the degree of vaginal wall organ motion during the entire course of therapy. RESULTS: The total motion of the fiducials center of mass (COM) was a median of 5.8 mm (range, 0.6-20.2 mm), and 95% of all COM positions fell within 15.7 mm of their original position. Directional margins of 3.1 mm along the right-left axis, 9.5 mm along the superoinferior axis, and of 12.1 mm along the anteroposterior axis encompassed the vaginal fiducials in 95% of treatments. Mean organ deformation for all patients was 3.9 mm, (range, 0-27.5 mm; standard deviation, 3.1 mm), with significant distortions of greater than 10 mm in 17% of secondary image sets. CONCLUSIONS: These data suggest a planning target volume margin of 16 mm will account for maximal organ motion in the majority of gynecologic patients undergoing postoperative pelvic IMRT, and it may be possible to incorporate directional motion into the planning target volume margin. PMID- 21640509 TI - Early significant tumor volume reduction after radiosurgery in brain metastases from renal cell carcinoma results in long-term survival. AB - PURPOSE: To retrospectively evaluate survival of patients with brain metastasis from renal cell carcinoma (RCC) after radiosurgery. PATIENTS AND METHODS: Between 1998 and 2010, 46 patients were treated with radiosurgery, and the total number of lesions was 99. The mean age was 58.9 years (range, 33-78 years). Twenty-six patients (56.5%) had a single brain metastasis. The mean tumor volume was 3.0 cm(3) (range, 0.01-35.1 cm(3)), and the mean marginal dose prescribed was 20.8 Gy (range, 12-25 Gy) at the 50% isodose line. A patient was classified into the good response group when the sum of the volume of the brain metastases decreased to less than 75% of the original volume at a 1-month follow-up evaluation using MRI. RESULTS: As of December 28, 2010, 39 patients (84.8%) had died, and 7 (15.2%) survived. The overall median survival time was 10.0 +/- 0.4 months (95% confidence interval, 9.1-10.8). After treatment, local tumor control was achieved in 72 (84.7%) of the 85 tumors assessed using MRI after radiosurgery. The good response group survived significantly longer than the poor-response group (median survival times of 18.0 and 9.0 months, respectively; p = 0.025). In a multivariate analysis, classification in the good-response group was the only independent prognostic factor for longer survival (p = 0.037; hazard ratio = 0.447; 95% confidence interval, 0.209-0.953). CONCLUSIONS: Radiosurgery seems to be an effective treatment modality for patients with brain metastases from RCC. The early significant tumor volume reduction observed after radiosurgery seems to result in long-term survival in RCC patients with brain metastases. PMID- 21640510 TI - Role of definitive radiation therapy in carcinoma of unknown primary in the abdomen and pelvis. AB - OBJECTIVES: Carcinoma of unknown primary (CUP) in the abdomen and pelvis is a heterogeneous group of cancers with no standard treatment. Considered by many to be incurable, these patients are often treated with chemotherapy alone. In this study, we determined the effectiveness of radiation therapy in combination with chemotherapy in patients with CUP in the abdomen and pelvis. PATIENTS AND METHODS: Medical records were reviewed for 37 patients with CUP treated with radiation therapy for disease located in the soft tissues and/or nodal basins of the abdomen and pelvis at the University of Texas M.D. Anderson Cancer between 2002 and 2009. All patients underwent chemotherapy, either before or concurrent with radiation therapy. Patients were selected for radiation therapy on the basis of histologic type, disease extent, and prior therapy response. Twenty patients underwent definitive radiation therapy (defined as radiation therapy targeting all known disease sites with at least 45 Gy) and 17 patients underwent palliative radiation therapy. Only 6 patients had surgical resection of their disease. Patient and treatment characteristics were extracted and the endpoints of local disease control, progression-free survival (PFS), overall survival (OS), and treatment-related toxicity incidence were analyzed. RESULTS: The 2-year PFS and OS rates for the entire cohort were 32% and 57%, respectively. However, in patients treated with definitive radiation therapy, the rates were 48% and 76%, and 7 patients lived more than 3 years after treatment with no evidence of disease progression. Nevertheless, radiation-associated toxicity was significant in this cohort, as 40% experienced Grade 2 or higher late toxicities. CONCLUSIONS: The use of definitive radiation therapy should be considered in selected patients with CUP in the soft tissues or nodal basins of the abdomen and pelvis. PMID- 21640511 TI - Is it time to tailor the prediction of radio-induced toxicity in prostate cancer patients? Building the first set of nomograms for late rectal syndrome. AB - PURPOSE: Development of user-friendly tools for the prediction of single-patient probability of late rectal toxicity after conformal radiotherapy for prostate cancer. METHODS AND MATERIALS: This multicenter protocol was characterized by the prospective evaluation of rectal toxicity through self-assessed questionnaires (minimum follow-up, 36 months) by 718 adult men in the AIROPROS 0102 trial. Doses were between 70 and 80 Gy. Nomograms were created based on multivariable logistic regression analysis. Three endpoints were considered: G2 to G3 late rectal bleeding (52/718 events), G3 late rectal bleeding (24/718 events), and G2 to G3 late fecal incontinence (LINC, 19/718 events). RESULTS: Inputs for the nomogram for G2 to G3 late rectal bleeding estimation were as follows: presence of abdominal surgery before RT, percentage volume of rectum receiving >75 Gy (V75Gy), and nomogram-based estimation of the probability of G2 to G3 acute gastrointestinal toxicity (continuous variable, which was estimated using a previously published nomogram). G3 late rectal bleeding estimation was based on abdominal surgery before RT, V75Gy, and NOMACU. Prediction of G2 to G3 late fecal incontinence was based on abdominal surgery before RT, presence of hemorrhoids, use of antihypertensive medications (protective factor), and percentage volume of rectum receiving >40 Gy. CONCLUSIONS: We developed and internally validated the first set of nomograms available in the literature for the prediction of radio induced toxicity in prostate cancer patients. Calculations included dosimetric as well as clinical variables to help radiation oncologists predict late rectal morbidity, thus introducing the possibility of RT plan corrections to better tailor treatment to the patient's characteristics, to avoid unnecessary worsening of quality of life, and to provide support to the patient in selecting the best therapeutic approach. PMID- 21640512 TI - Prostate-specific antigen velocity before and after elimination of factors that can confound the prostate-specific antigen level. AB - PURPOSE: Prostate-specific antigen (PSA) velocity, like PSA level, can be confounded. In this study, we estimated the impact that confounding factors could have on correctly identifying a patient with a PSA velocity >2 ng/ml/y. METHODS AND MATERIALS: Between 2006 and 2010, a total of 50 men with newly diagnosed PC comprised the study cohort. We calculated and compared the false-positive and false-negative PSA velocity >2 ng/ml/y rates for all men and those with low-risk disease using two approaches to calculate PSA velocity. First, we used PSA values obtained within 18 months of diagnosis; second, we used values within 18 months of diagnosis, substituting the prebiopsy PSA for a repeat, nonconfounded PSA that was obtained using the same assay and without confounders. RESULTS: Using PSA levels pre-biopsy, 46% of all men had a PSA velocity >2 ng/ml/y; whereas this value declined to 32% when substituting the last prebiopsy PSA for a repeat, nonconfounded PSA using the same assay and without confounders. The false positive rate for PSA velocity >2 ng/ml/y was 43% as compared with a false negative rate of PSA velocity >2 ng/ml/y of 11% (p = 0.0008) in the overall cohort. These respective values in the low-risk subgroup were 60% and 16.7% (p = 0.09). CONCLUSION: This study provides evidence to explain the discordance in cancer-specific outcomes among groups investigating the prognostic significance of PSA velocity >2 ng/ml/y, and highlights the importance of patient education on potential confounders of the PSA test before obtaining PSA levels. PMID- 21640513 TI - Curative treatment of Stage I non-small-cell lung cancer in patients with severe COPD: stereotactic radiotherapy outcomes and systematic review. AB - OBJECTIVES: Patients with severe chronic obstructive pulmonary disease (COPD) have a high risk of lung cancer and of postsurgical complications. We studied outcomes after stereotactic body radiotherapy (SBRT) in patients with severe COPD, as defined by Global Initiative for Chronic Obstructive Lung Disease (GOLD) criteria, and performed a systematic review of the literature on outcomes after SBRT or surgery in these patients. METHODS: A single-institution cohort of 176 patients with COPD GOLD III-IV and Stage I non-small-cell lung cancer (NSCLC) treated with SBRT was evaluated. A systematic review identified studies reporting outcomes after SBRT or surgery for Stage I NSCLC in patients with GOLD III-IV or a predicted postoperative forced expiratory volume in 1 second (FEV1) of <=40%. RESULTS: In the single-institution cohort, median follow-up was 21 months and median overall survival (OS) was 32 months. Actuarial 3-year local control was 89%, and 1- and 3-year OS were 79% and 47%, respectively. COPD severity correlated with OS (p = 0.01). The systematic review identified four other studies (two surgical, two SBRT, n = 196 patients). SBRT studies were published more recently and included older patients than surgical studies. Mean 30-day mortality was 0% post-SBRT and 10% after surgery. Local or locoregional control was high (>=89%) after both treatments. Post-SBRT, actuarial OS was 79-95% at 1 year and 43-70% at 3 years. Postsurgical actuarial OS was 45-86% at 1 year and 31 66% at 3 years. CONCLUSIONS: SBRT and surgery differ in risk of 30-day mortality in patients with severe COPD. Despite the negative selection of SBRT patients, survival at 1 and 3 years is comparable between the two treatments. PMID- 21640514 TI - Localized orbital mucosa-associated lymphoma tissue lymphoma managed with primary radiation therapy: efficacy and toxicity. AB - PURPOSE: To evaluate the clinical outcomes and late effects of radiation therapy (RT) in localized primary orbital mucosa-associated lymphoma tissue (MALT) lymphoma (POML). METHODS AND MATERIALS: From 1989 to 2007, 89 patients with Stage IE POML received RT. The median age was 56 years old. Sites involved conjunctiva (59 patients [66%]), lacrimal gland (20 patients [23%]), and soft tissue (10 patients [11%]). Megavoltage beam(s) was used in 91%, electrons in 7%, and orthovoltage in 2% of cases. The dose given was 25 Gy in 97% and 30 Gy in 3% of patients. Lens shielding was possible in 57% of patients. RESULTS: The median follow-up was 5.9 years. Complete response or unconfirmed complete response was seen in 88 patients (99%). Relapse occurred in 22 patients (25%). First relapse sites were local (2 patients [9%]), in the contralateral orbit (5 patients [23%]), and distant (15 patients [68%]). The 7-year overall survival (OS), cause specific survival (CSS), relapse-free survival (RFS), and local control (LC) rates were 91%, 96%, 64%, and 97%, respectively. Radiation-related late sequelae were documented in 40 patients (45%). Cataracts were observed in 22 patients (Grade 1 in 2 patients; Grade 3 in 20 patients). The incidence of Grade 3 cataract at 7 years was 25%. Other late sequelae (n = 28) were dry eye(s) (22 patients [Grade 1 in 14 patients; Grade 2 in 2 patients; Grade 3 in 2 patients; n/s in 4 patients), keratitis (3 patients), macular degeneration/cystoid edema (2 patients), and vitreous detachment (1 patient). Five patients developed Grade 3 noncataract late effects. Lens shielding reduced the incidence of Grade 3 cataract and all Grade >=2 late sequelae. Seventeen patients (16 with cataracts) underwent surgery; 23 patients were treated conservatively. The outcome for managing late effects was generally successful, with 30 patients completely improved, and 9 patients with persisting late sequelae (10%). CONCLUSIONS: POML responds favorably to moderate doses of RT but results in significant late morbidity. The majority of late effects were successfully managed. Lens shielding reduced the risk of cataracts and other late sequelae. PMID- 21640515 TI - Dosimetric advantage of intensity-modulated radiotherapy for whole ventricles in the treatment of localized intracranial germinoma. AB - PURPOSE: To investigate the dosimetric advantage of intensity-modulated radiotherapy (IMRT) for whole ventricles (WV) in patients with a localized intracranial germinoma receiving induction chemotherapy. METHODS AND MATERIALS: Data from 12 consecutive patients with localized intracranial germinomas who received induction chemotherapy and radiotherapy were used. Four-field coplanar three-dimensional conformal radiotherapy (3D-CRT) and seven-field coplanar IMRT plans were created. In both plans, 24 Gy was prescribed in 12 fractions for the planning target volume (PTV) involving WV and tumor bed. In IMRT planning, optimization was conducted to reduce the doses to the organs at risk (OARs) as much as possible, keeping the minimum dose equivalent to that of 3D-CRT. The 3D CRT and IMRT plans were compared in terms of the dose-volume statistics for target coverage and the OARs. RESULTS: IMRT significantly increased the percentage volume of the PTV receiving 24 Gy compared with 3D-CRT (93.5% vs. 84.8%; p = 0.007), while keeping target homogeneity equivalent to 3D-CRT (p = 0.869). The absolute percentage reduction in the irradiated volume of the normal brain receiving 100%, 75%, 50%, and 25% of 24 Gy ranged from 0.7% to 16.0% in IMRT compared with 3D-CRT (p < 0.001). No significant difference was observed in the volume of the normal brain receiving 10% and 5% of 24 Gy between IMRT and 3D CRT. Conformation number was significantly improved in IMRT (p < 0.001). For other OARs, the mean dose to the cochlea was reduced significantly in IMRT by 22.3% of 24 Gy compared with 3D-CRT (p < 0.001). CONCLUSIONS: Compared with 3D CRT, IMRT for WV improved the target coverage and reduced the irradiated volume of the normal brain in patients with intracranial germinomas receiving induction chemotherapy. IMRT for WV with induction chemotherapy could reduce the late side effects from cranial irradiation without compromising control of the tumor. PMID- 21640516 TI - Radiotherapy of painful vertebral hemangiomas: the single center retrospective analysis of 137 cases. AB - PURPOSE: An evaluation of dose-response relationship and an attempt to define predictive factors. METHODS AND MATERIALS: A total of 137 cases of painful vertebral hemangioma irradiations (101 patients). Fraction dose (fd) varied from 2 to 15 Gy (123 fractionated and 14 radiosurgical treatments), and total dose (TD) from 8 to 30 Gy (111 cases irradiated with fd of 2 GY to TD of 24 Gy). We evaluated pain relief, changes in analgesic requirements, and reossification. RESULTS: Means of pain relief 1, 6, 12, and 18 months after radiotherapy (defined as a decrease of primary pain level expressed in percent) were 60.5%, 65.4%, 68.3%, and 78.4%, respectively. Proportion of patients with no need for analgesics and patients using tramadol were 39%, 40%, 44%, 57%, and 20%, 17%, 22%, and 11% in these times. The proportion of patients experiencing complete/partial pain relief changed from 36/48% 1 month, to 64/22% 1.5 years after radiotherapy. No impact of radiotherapy on reossification was found. The positive impact of fd and TD increase for analgesics uptake reduction and pain relief was found. An increase of the fd by 1 Gy results in 27% chance of analgesics uptake reduction and 3.8% reduction of pain, whereas 14% analgesics uptake reduction and 2.2% of pain reduction in case of the TD. The predictive factors improving results were found: female gender, older age, better performance states (the chance of the lower analgesic treatment decreases over 2.5 times in comparison to the higher Zubrod degree), bigger Hb concentration, shorter symptoms duration and lower analgesics uptake before radiotherapy. CONCLUSIONS: The obtained data support the efficacy of radiotherapy in improving pain secondary to vertebral hemangioma, with the degree of pain amelioration being related to increasing fd and TD. The positive predictive factors were defined: female gender, older age, better performance status, increased Hb concentration, shorter symptoms duration, and lower analgesics uptake before radiotherapy. PMID- 21640517 TI - Primary Gleason grade 4 impact on biochemical recurrence after permanent interstitial brachytherapy in Japanese patients with low- or intermediate-risk prostate cancer. AB - PURPOSE: To reveal a predictive factor for biochemical recurrence (BCR) after permanent prostate brachytherapy (PPB) using iodine-125 seed implantation in patients with localized prostate cancer classified as low or intermediate risk based on National Comprehensive Cancer Network (NCCN) guidelines. METHODS AND MATERIALS: From January 2004 to December 2009, 414 consecutive Japanese patients with clinically localized prostate cancer classified as low or intermediate risk based on the NCCN guidelines were treated with PPB. The clinical factors including pathological data reviewed by a central pathologist and follow-up data were prospectively collected. Kaplan-Meier and Cox regression analyses were used to assess the factors associated with BCR. RESULTS: Median follow-up was 36.5 months. The 2-, 3-, 4-, and 5-year BCR-free rates using the Phoenix definition were 98.3%, 96.0%, 91.6%, and 87.0%, respectively. On univariate analysis, the Gleason score, especially primary Gleason grade 4 in biopsy specimens, was a strong predicting factor (p < 0.0001), while age, initial prostate-specific antigen (PSA) level, T stage, and minimal dose delivered to 90% of the prostate volume (D90) were insignificant. Multivariate analysis indicated that a primary Gleason grade 4 was the most powerful prognostic factor associated with BCR (hazard ratio = 6.576, 95% confidence interval, 2.597-16.468, p < 0.0001). CONCLUSIONS: A primary Gleason grade 4 carried a worse BCR prognosis than the primary grade 3 in patients treated with PPB. Therefore, the indication for PPB in patients with a Gleason sum of 4 + 3 deserves careful and thoughtful consideration. PMID- 21640518 TI - Concurrent hyperfractionated radiation therapy and chemotherapy in locally advanced (Stage III) non-small-cell lung cancer: single institution experience with 600 patients. AB - PURPOSE: Our institutional experience with the use of hyperfractionated radiation therapy (RT) alone or concurrently with chemotherapy (RT-CHT) in Stage III non small-cell lung cancer was reviewed. METHODS AND MATERIALS: Three phase III and two phase II studies included a total of 600 patients. Hyperfractionated RT alone was given to 127 patients, and hyperfractionated RT-CHT was given to 473 patients. RT doses were 64.8 Gy and 69.6 Gy (using 1.2 Gy twice daily) and 67.6 Gy (using 1.3 Gy twice daily). CHT consisted of concurrent administration of carboplatin and etoposide to 409 patients and concurrent administration of carboplatin and paclitaxel to 64 patients. RESULTS: The median survival times were 19 months, 21 months, and 12 months for all, RT-CHT, and RT-only patients, respectively. The survival difference between the RT-CHT and RT group was significant (p < 0.0001). Four-year rates of local progression-free survival (LPFS) and distant metastasis-free survival (DMFS) were 29% and 35%, respectively, for the entire group. The RT-CHT group had significantly better LPFS rates than the RT group (31% for the RT-CHT group vs. 16% for the RT group; p = 0.0015) but not DMFS rates (36% for the RT-CHT group vs. 36% for the RT group, p = 0.0571). Acute high-grade esophagitis, pneumonitis, and hematological toxicities were seen most frequently and in 11%, 9%, and 12% of patients, respectively. Late high-grade esophageal and bronchopulmonary toxicity were each seen in 6% of patients. CONCLUSIONS: Compared to the majority of existing phase II and III studies, this study reconfirmed the excellent results achieved with concurrent RT-CHT, including low toxicity. Concurrent RT-CHT results in survival benefit primarily by increasing LPFS, not DMFS. PMID- 21640519 TI - Impact of hypoxia on the metastatic potential of human prostate cancer cells. AB - PURPOSE: Intratumoral hypoxia is known to be associated with radioresistance and metastasis. The present study examined the effect of acute and chronic hypoxia on the metastatic potential of prostate cancer PC-3, DU145, and LNCaP cells. METHODS AND MATERIALS: Cell proliferation and clonogenicity were tested by MTT assay and colony formation assay, respectively. "Wound-healing" and Matrigel-based chamber assays were used to monitor cell motility and invasion. Hypoxia-inducible factor 1 alpha (HIF-1alpha) expression was tested by Western blot, and HIF-1-target gene expression was detected by real-time polymerase chain reaction. Secretion of matrix metalloproteinases (MMPs) was determined by gelatin zymography. RESULTS: When PC-3 cells were exposed to 1% oxygen (hypoxia) for various periods of time, chronic hypoxia (>=24 h) decreased cell proliferation and induced cell death. In contrast, prostate cancer cells exposed to acute hypoxia (<=6 h) displayed increased motility, clonogenic survival, and invasive capacity. At the molecular level, both hypoxia and anoxia transiently stabilized HIF-1alpha. Exposure to hypoxia also induced the early expression of MMP-2, an invasiveness-related gene. Treatment with the HIF-1 inhibitor YC-1 attenuated the acute hypoxia-induced migration, invasion, and MMP-2 activity. CONCLUSIONS: The length of oxygen deprivation strongly affected the functional behavior of all three prostate cancer cell lines. Acute hypoxia in particular was found to promote a more aggressive metastatic phenotype. PMID- 21640520 TI - Fifteen-year radiotherapy outcomes of the randomized PORTEC-1 trial for endometrial carcinoma. AB - PURPOSE: To evaluate the very long-term results of the randomized Post Operative Radiation Therapy in Endometrial Carcinoma (PORTEC)-1 trial for patients with Stage I endometrial carcinoma (EC), focusing on the role of prognostic factors for treatment selection and the long-term risk of second cancers. PATIENTS AND METHODS: The PORTEC trial (1990-1997) included 714 patients with Stage IC Grade 1 2 or Stage IB Grade 2-3 EC. After surgery, patients were randomly allocated to external-beam pelvic radiotherapy (EBRT) or no additional treatment (NAT). Analysis was by intention to treat. RESULTS: 426 patients were alive at the date of analysis. The median follow-up time was 13.3 years. The 15-year actuarial locoregional recurrence (LRR) rates were 6% for EBRT vs. 15.5% for NAT (p < 0.0001). The 15-year overall survival was 52% vs. 60% (p = 0.14), and the failure free survival was 50% vs. 54% (p = 0.94). For patients with high-intermediate risk criteria, the 15-year overall survival was 41% vs. 48% (p = 0.51), and the 15-year EC-related death was 14% vs. 13%. Most LRR in the NAT group were vaginal recurrences (11.0% of 15.5%). The 15-year rates of distant metastases were 9% vs. 7% (p = 0.25). Second primary cancers had been diagnosed over 15 years in 19% of all patients, 22% vs. 16% for EBRT vs. NAT (p = 0.10), with observed vs. expected ratios of 1.6 (EBRT) and 1.2 (NAT) compared with a matched population (p = NS). Multivariate analysis confirmed the prognostic significance of Grade 3 for LRR (hazard ratio [HR] 3.4, p = 0.0003) and for EC death (HR 7.3, p < 0.0001), of age >60 (HR 3.9, p = 0.002 for LRR and 2.7, p = 0.01 for EC death) and myometrial invasion >50% (HR 1.9, p = 0.03 and HR 1.9, p = 0.02). CONCLUSIONS: The 15-year outcomes of PORTEC-1 confirm the relevance of HIR criteria for treatment selection, and a trend for long-term risk of second cancers. EBRT should be avoided in patients with low- and intermediate-risk EC. PMID- 21640521 TI - The Maillard reaction--illicite (bio)chemistry in tissues and food. AB - We present a review of our early work on the Maillard reaction, at the interface of food chemistry and tissue biochemistry, as well as the reinterpretation of our early findings in the light of recent advances in the chemistry of the involved reactions. These concern specifically the role of lower aldehydes, produced during the glycolytic pathways and especially acetaldehyde. We also review some of our recent findings on the cytotoxic and genotoxic aspect of these "illicit" organic reactions, taking place in tissues (and also in food products) besides the genetically "programmed" metabolic pathways. Some recent results in organic pharmaceutical chemistry confirm the potential importance of the reviewed reactions both in food chemistry and in tissues as well as the pathological importance of reactions taking place in tissues. PMID- 21640522 TI - Predicting rapid DUI recidivism using the Driver Risk Inventory on a state-wide sample of Floridian DUI offenders. AB - BACKGROUND: This project examined the ability of the popular DUI/DWI offender assessment instrument, the Driver Risk Inventory (DRI; Behavior Data Systems, Ltd., 1985), to identify short-term DUI recidivists in a sample of Floridian DUI offenders who were charged with DUI between January 1st, 2008 and December 31st, 2009. The DRI provides a number of behavioral risk scales, DSM-IV substance abuse and dependence classifications, as well as measurement of demographic and criminal history characteristics. METHODS: Cox proportional hazards models were used to identify the demographic, criminal history, and behavioral characteristics most closely associated with the risk of rapid DUI recidivism. Follow-up analyses including ROC curves were used to further examine the ability of the DRI to identify short-term DUI recidivists. RESULTS: In the final model controlling for all variables, the DRI driver risk scale was the single strongest predictor of rapid DUI recidivism. The DSM-IV substance abuse and dependence classifications were also significant predictors of DUI recidivism. A number of the DRI risk scales and the DSM-IV classifications exhibited significant predictive validity and exhibited sensitivity in identifying recidivists similar to other popular DUI offender assessment instruments. CONCLUSIONS: The DRI provides useful identification of DUI recidivists in a sample able to capture only the most rapid DUI recidivists. The results of this research warrant further examination of the DRI's ability to identify DUI recidivists using longer intervals of time between DUI arrests. PMID- 21640523 TI - Mycoplasma ovis in captive cervids: prevalence, molecular characterization and phylogeny. AB - Hemotrophic mycoplasmas (hemoplasmas) are bacteria that attach to red blood cells of mammals, leading to acute and/or subclinical disease in infected animals. It has been suggested that Mycoplasma ovis, a hemoplasma that infects sheep and goats worldwide, may also infect deer. The aim of this study was to evaluate whether South American deer are infected with M. ovis. EDTA-anticoagulated blood samples from a herd of 32 captive South American deer were collected. DNA extraction of blood samples was performed followed by PCR amplification of the 16S and 23S rRNA genes, and sequencing of products. Using M. ovis PCR, 27/31 (87%) were positive, including 21/22 Mazama nana; 2/3 Mazama americana and 4/6 Blastocerus dichotomus. Sequencing of the nearly entire 16S rRNA gene of 26/27 positive samples showed 98.2-98.8% identity to M. ovis of sheep (GenBank, AF338268) and 98.6-99.4% identity to M. ovis-like of a fawn (FJ824847); the 23S rRNA gene from one of these isolates and the fawn's had 97.6% identity. The remaining isolate had just 94.9% identity to the 16S rRNA gene of M. ovis and only 89.4% identity to the 23S rRNA gene of the fawn's M. ovis. This is the first report of M. ovis in captive South American deer, revealing a high prevalence of hemoplasma infection in these animals. PMID- 21640524 TI - The bovine herpesvirus type 1 envelope protein Us9 acidic domain is crucial for anterograde axonal transport. AB - In this study, we examined the functional role of bovine herpesvirus type 1 (BHV 1) Us9 acidic domain residues 83-90 in the anterograde axonal transport of the virus in calves (natural host), rabbits, and in cultured neurons. A mutant virus strain lacking Us9 residues 83-90 (BHV-1 Us9 Delta83-90) and the rescued virus (BHV-1 Us9 R83-90) replicated efficiently in the nasal and ocular epithelium during primary infection and established latency in the trigeminal ganglia (TG). However, upon reactivation from latency, only the BHV-1 Us9 R83-90 virus was detected in nasal and ocular swabs of animals. In compartmentalized, rabbit primary dorsal root ganglia (DRG) neuron cultures, the Us9-deleted BHV-1, BHV-1 Us9 Delta83-90 and BHV-1 Us9 R83-90 viruses were transported efficiently in the retrograde direction. However, only the BHV-1 Us9 R83-90 virus was transported in an anterograde direction. These studies suggested that the Us9 acidic domain residues located between 83 and 90 were required for axonal anterograde transport. PMID- 21640525 TI - Neurodevelopmental and neurofunctional outcomes in children with congenital diaphragmatic hernia. AB - The objective of this review was to provide a critical overview of our current understanding on the neurocognitive, neuromotor, and neurobehavioral development in congenital diaphragmatic hernia (CDH) patients, focusing on three interrelated clinical issues: (1) comprehensive outcome studies, (2) characterization of important predictors of adverse outcome, and (3) the pathophysiological mechanism contributing to neurodevelopmental disabilities in infants with CDH. Improved survival for CDH has led to an increasing focus on longer-term outcomes. Neurodevelopmental dysfunction has been recognized as the most common and potentially most disabling outcome of CDH and its treatment. While increased neuromotor dysfunction is a common problem during infancy, behavioral problems, hearing impairment and quality of life related issues are frequently found in older children and adolescence. Intelligence appears to be in the low normal range. Patient and disease specific predictors of adverse neurodevelopmental outcome have been defined. Imaging studies have revealed a high incidence of structural brain abnormalities. An improved understanding of the pathophysiological pathways and the neurodevelopmental consequences will allow earlier and possibly more targeted therapeutic interventions. Continuous assessment and follow-up as provided by an interdisciplinary team of medical, surgical and developmental specialists should become standard of care for all CDH children to identify and treat morbidities before additional disabilities evolve and to reduce adverse outcomes. PMID- 21640526 TI - Ages and Stages Questionnaires: feasibility of postal surveys for child follow up. AB - BACKGROUND: The Ages and Stages Questionnaire (ASQ), completed by parents and caregivers, has been shown to be an accurate tool for screening children who need further developmental assessment. AIMS: To assess the feasibility of using the French Canadian translation of the ASQ in an epidemiological cohort of children from the French general population. STUDY DESIGN: Follow-up study by postal questionnaire at 12 and 36 months, using the ASQ. SUBJECTS: 339 French families recruited at the birth of their child in 2006 in two hospitals in the Paris suburbs. OUTCOME MEASURE: Response rates and French ASQ results at 12 and 36 months. The ASQ was scored as indicated in the manual. RESULTS: A high response rate of 79% was observed at the children's 1st and 3rd birthdays. Parents were enthusiastic about participating; half of them wrote comments on the questionnaires, most of them positive. Low scores at the 12-month assessment were associated with birth characteristics such as prematurity and transfer to the neonatology unit after birth, whereas at 36 months they tended to be associated with both birth and family socio-demographic characteristics. CONCLUSIONS: Use of the French ASQ in a research cohort appears feasible as response rates were high. Moreover, known links between child development measured by ASQ and birth and social characteristics were observed. However, further French studies are needed to understand differences observed in 12-month ASQ gross motor scores compared with US norms. For research purposes, further analysis of the ASQ in innovative, quantitative approaches, is needed. PMID- 21640527 TI - Characterization of epididymal spermatozoa motility rate, morphology and longevity of springbok (Antidorcas marsupialis), impala (Aepyceros melampus) and blesbok (Damaliscus dorcus phillipsi): pre- and post-cryopreservation in South Africa. AB - Cryopreservation of antelope epididymal spermatozoa could play a vital role in future breeding by developing a successful protocol for cryo-conserving them. The aim of this study was to characterize morphology, motility rates and longevity of epididymal spermatozoa from springbok, impala and blesbok. Cauda epididymal spermatozoa were collected post-mortem from both testicles of free-ranging springbok (n=18), impala (n=21) and blesbok (n=21), and divided into two groups (pre- and post-cryopreservation). Spermatozoa were cryopreserved in Biladyl supplemented with 20% egg yolk and 7% glycerol under field conditions. Pre-freeze and post-thaw sperm quality was evaluated. The longevity of thawed spermatozoa was evaluated under culture conditions that support domestic cattle in vitro fertilization. There was a significant difference between pre-freeze and post thaw sperm motility index (SMI) (p<0.05), plasma membrane integrity (p<0.05) and acrosome integrity (p<0.05) for all species. Post-thaw SMI and plasma membrane integrity were comparable between species (p>0.05). The effects of cryopreservation on sperm cell morphology differed between species and between specific abnormal morphology. Blesbok had the least abnormalities in post-thaw spermatozoa. Cryopreservation substantially reduced the survivability and motility rates of antelope species. Blesbok spermatozoa tolerated cryopreservation and thawing process better than impala and springbok. The antelope cauda epididymal sperm maintained viability and acrosome integrity for at least 4h following incubation under conditions that support domestic cattle in vitro fertilization (IVF) with a decline in longevity over time across species however; species responded differently over time in terms of plasma membrane integrity and acrosome integrity. The antelope species may have different in vitro culture requirements, indicating differences in sperm physiology between the species. This research could contribute species-specific protocol development for IVF thus promoting ex-situ conservation strategies of African antelope species in South Africa. PMID- 21640528 TI - Microisolation and microcloning of bovine X-chromosomes for identification of sorted buffalo (Bubalus bubalis) spermatozoa. AB - Flow-cytometry sorting technology has been successfully used to separate the X- and Y-chromosome bearing spermatozoa for production of sex-preselected buffalo. However, an independent technique should be employed to validate the sorting accuracy. In the present study, X-chromosomes of bovine were micro-dissected from the metaphase spreads by using glass needles. Then X-chromosomes were then amplified by PCR and labelled with Cy3-dUTP for use as a probe in hybridization of the unsorted and sorted buffalo spermatozoa -chromosome. The results revealed that 47.7% (594/1246) of the unsorted buffalo spermatozoa were positive for X- chromosome probe, which was conformed to the sex ratio in buffalo (X:Y spermatozoa=1:1); 9.6% (275/2869) of the Y-sorted buffalo spermatozoa and 86.1% (1529/1776) of the X-sorted buffalo spermatozoa showed strong X-chromosome FISH signals. Flow cytometer re-analysis revealed that the proportions of X- and Y bearing spermatozoa in the sorted X and Y semen was 89.6% and 86.7%, respectively. There were no significant differences between results assayed by flow-cytometry re-analysis and by FISH in this study. In conclusion, FISH probe derived from bovine X- chromosomes could be used to verify the purity of X and Y sorted spermatozoa in buffalo. PMID- 21640529 TI - Seven cases of fatal aconite poisoning: forensic experience in China. AB - This paper presents seven fatal cases of aconite poisoning encountered in the Tongji Center for Medicolegal Expertise in Hubei (TCMEH), China, from 1999 to 2008 retrospectively. In six of the cases, deaths occurred after drinking homemade medicated liquor containing aconite, and in one case death was due to ingestion of traditional Chinese medication containing aconite. Forensic autopsy and pathological examinations ruled out the presence of physical trauma or life threatening diseases. Diagnosis of aconite poisoning was made after postmortem toxicological analysis. Animal experiment was performed in one case demonstrating that the medicated liquor could cause death rapidly. We present the autopsy and histopathological findings, toxicological analysis, and results of animal experiment done on samples from those seven cases. As an important herbal Chinese medicine, Aconitum species deserve special attention, especially because it contains poisonous alkaloids. PMID- 21640530 TI - Surprising hair analysis results following acute carbofuran intoxication. AB - We present two non fatal cases of intoxication with carbofuran (CBF) documented by hair analysis. Carbofuran and 3-hydroxycarbofuran (OHCBF, its main metabolite) hair concentrations were determined using a liquid chromatography-tandem mass spectrometry method. The obtained results were surprising if we consider several hair analyses previously published and based on a theory of the presence of xenobiotic in the only segment that comprised its intake. Among the two intoxication cases, we noticed the presence of CBF and OHCBF in hair segments corresponding to 45 days before, and more than 100 days after, the day of intoxication. Additionally, repeated hair samplings and subsequent analysis revealed a decrease of the carbofuran's concentration during the hair life. PMID- 21640531 TI - Operational benefits and challenges of the use of fingerprint statistical models: a field study. AB - Research projects aimed at proposing fingerprint statistical models based on the likelihood ratio framework have shown that low quality finger impressions left on crime scenes may have significant evidential value. These impressions are currently either not recovered, considered to be of no value when first analyzed by fingerprint examiners, or lead to inconclusive results when compared to control prints. There are growing concerns within the fingerprint community that recovering and examining these low quality impressions will result in a significant increase of the workload of fingerprint units and ultimately of the number of backlogged cases. This study was designed to measure the number of impressions currently not recovered or not considered for examination, and to assess the usefulness of these impressions in terms of the number of additional detections that would result from their examination. PMID- 21640532 TI - Effects of synthetic cannabinoids on electroencephalogram power spectra in rats. AB - Several synthetic cannabinoids have recently been distributed as psychoactive adulterants in many herbal products on the illegal drug market around the world. However, there is little information on pharmacology and toxicology of such compounds. Although Delta(9)-tetrahydrocannabinol (Delta(9)-THC), a psychoactive cannabinoid of marijuana, was reported to affect electroencephalograms (EEG) of rats, the effects of synthetic cannabinoids are unknown. We examined the pharmacological activities of three synthetic cannabinoids; cannabicyclohexanol (CCH), CP-47,497 and JWH-018; by analyzing EEG power spectra and locomotor activity after intraperitoneal administration to rats and compared them with those of Delta(9)-THC. The three compounds significantly increased the EEG power in the frequency range of 5.0-6.0 Hz for the first 3h, while Delta(9)-THC decreased the power spectra in the wide range of 7.0-20.0 Hz during the first hour. These results indicate that the effect of the three compounds on EEG is different from that of Delta(9)-THC. Additionally, CCH, CP-47,497 and JWH-018 significantly decreased the locomotor activity for 11.5h, 11h and 4.5h, respectively, after administration which was longer than that of Delta(9)-THC (3.5h). Furthermore, all three compounds significantly reduced the total amounts of locomotor activity during a 3-h, 6-h and 12-h period after injection, whereas no statistical difference was observed for the Delta(9)-THC injection. Among the three compounds, CCH and CP-47,497 exerted a longer duration of the change in the EEG power spectra and suppression of the locomotor activity than JWH-018. PMID- 21640533 TI - A case of midgut malrotation presenting as multiple afebrile seizures. AB - Provoked seizures are a common pediatric problem and can be caused by multiple etiologies. However, there have been few reports of benign infantile seizures associated with gastrointestinal tract abnormality. We report on a 3-month-old male infant who presented with multiple afebrile seizures, EEG abnormalities and a midgut malrotation. Surgical treatment of the malrotation resulted in seizure resolution and EEG normalization. This case emphasizes that patients presenting with multiple seizures and symptoms of gastroenteritis should be evaluated for abnormalities of the gastrointestinal tract. PMID- 21640534 TI - Investigation and management of an imipenem-resistant oxa-23 Acinetobacter baumannii outbreak in an intensive care unit. AB - OBJECTIVES: The study objectives were to describe the investigation and management of an imipenem-resistant Acinetobacter baumannii outbreak that occurred in the 15-bed ICU of a tertiary care teaching hospital (Brest, France), during the summer 2008. PATIENTS AND METHODS: Patients harboring an imipenem resistant A. baumannii strain were defined as case patients. We described case occurrence and steps taken to control the outbreak: contact isolation, reinforcement of hygiene procedures, unit shutdown decision, unit disinfection, and reopening. We also made a case control study and a cost analysis of the outbreak management. RESULTS: During a 10-day period, five patients were positive for a single clone of imipenem-resistant oxa-23 A. baumannii. Four patients presented with ventilation-acquired pneumonia and one was asymptomatic. The first two patients died one day after the first swab which led to the identification of A. baumannii. No additional case was noted in the ICU or in other hospital units after deciding to close the ICU. The cost of outbreak management was estimated at 264,553 euros. The case control study identified several factors associated with infection or colonization: length of stay in the ICU, chronic respiratory disease, number of previous antibiotic classes used, duration of ventilation, prone position, echocardiography, and presence of a nasogastric tube. CONCLUSION: This outbreak occurred during the summer period requiring the shutdown of the ICU and inducing a considerable cost. Rapid reactions of the ICU staff during the outbreak enabled to limit the epidemic. PMID- 21640535 TI - Which concentration to choose in dual flow cardiac CT?: dual flow cardiac CT. AB - PURPOSE: An extensive number of protocols have been suggested to allow for functional diagnostics; however, no data is available about the minimal amount of contrast medium to achieve reliable imaging properties. None of the plethora of existing studies report a rational why the specific concentration was chosen. MATERIALS AND METHODS: A total of 40 patients were included in this prospective, controlled study. They were divided up into four equal groups getting a different concentration (10%, 20%, 30% or 40%) of a second contrast medium bolus. Corresponding septal and right ventricular ROIs were compared. A visual score was established. Coronary attenuation was measured in the right and left coronary artery. Streak artifacts in the right atrium/ventricle were assessed. RESULTS: In the 10% contrast medium (CM) group only in 5/10 (50%) patients full septal delineation was reached. In all other groups full septal visualization was obtained. No group showed a relevant difference of mean density measured in HU units of the left ventricle or the coronary arteries. All study groups except of group 1 (10% CM) showed streak artifacts in the right atrium. CONCLUSION: The dual flow protocol with a minimum concentration of 20% improves septal visualization as basis for left ventricular functional assessment, however, does not allow for reliable right ventricular or atrial visualization. There is no significant difference between the different concentration protocols in terms of coronary attenuation. PMID- 21640536 TI - Effect of contrast material on image noise and radiation dose in adult chest computed tomography using automatic exposure control: a comparative study between 16-, 64- and 128-slice CT. AB - PURPOSE: To determine the difference in radiation dose between non-enhanced (NECT) and contrast-enhanced (CECT) chest CT examinations contributed by contrast material with different scanner generations with automatic exposure control (AEC). METHODS & MATERIALS: Each 42 adult patients received a NECT and CECT of the chest in one session on a 16-, 64- or 128-slice CT scanner with the same scan protocol settings. However, AEC technology (Care Dose 4D, Siemens) underwent upgrades in each of the three scanner generations. DLP, CTDIvol and image noise were compared. RESULTS: Although absolute differences in image noise were very small and ranged between 10 and 13 HU for NECT and CECT in median, the differences in image noise and dose (DLP: 16-slice:+2.8%; 64-slice:+3.9%; 128 slice:+5.6%) between NECT and CECT were statistically significant in all groups. Image noise and dose parameters were significantly lower in the most recent 128 slice CT generation for both NECT and CECT (DLP: 16-slice:+35.5-39.2%; 64 slice:+6.8-8.5%). CONCLUSION: The presence of contrast material lead to an increase in dose for chest examinations in three CT generations with AEC. Although image noise values were significantly higher for CECT, the absolute differences were in a range of 3 HU. This can be regarded as negligible, thus indicating that AEC is able to fulfill its purpose of maintaining image quality. However, technological developments lead to a significant reduction of dose and image noise with the latest CT generation. PMID- 21640537 TI - Methylobacillus arboreus sp. nov., and Methylobacillus gramineus sp. nov., novel non-pigmented obligately methylotrophic bacteria associated with plants. AB - Two newly isolated obligate methanol-utilizing bacteria (strains Iva(T) and Lap(T)) with the ribulose monophosphate pathway of C(1) assimilation are described. The isolates are strictly aerobic, Gram negative, asporogenous, motile rods multiplying by binary fission, mesophilic and neutrophilic, synthesize indole-3-acetate. The prevailing cellular fatty acids are straight-chain saturated C(16:0) and unsaturated C(16:1) acids. The major ubiquinone is Q-8. The predominant phospholipids are phosphatidylethanolamine, phosphatidylglycerol and cardiolipin. Ammonia is assimilated by glutamate dehydrogenase. The DNA G+C contents of strains Iva(T) and Lap(T) are 54.0 and 50.5mol% (T(m)), respectively. Based on 16S rRNA gene sequence analysis and DNA-DNA relatedness (38-45%) with type strains of the genus Methylobacillus, the novel isolates are classified as the new species of this genus and named Methylobacillus arboreus Iva(T) (VKM B 2590(T), CCUG 59684(T), DSM 23628(T)) and Methylobacillus gramineus Lap(T) (VKM B 2591(T), CCUG 59687(T), DSM 23629(T)). The GenBank accession numbers for the 16S rRNA gene and mxaF gene sequences of the strains Iva(T) and Lap(T) are GU937479, GU937478 and HM030736, HM030735, respectively. PMID- 21640538 TI - Bacillus locisalis sp. nov., a new haloalkaliphilic species from hypersaline and alkaline lakes of China, Kenya and Tanzania. AB - A polyphasic taxonomic study was performed on seven Bacillus-like bacteria isolated from three hypersaline and alkaline lakes located in China, Kenya and Tanzania. All strains were moderately halophilic and alkaliphilic, Gram positive, motile rods. The DNA G+C content from the seven isolates ranged from 42.2 to 43.4mol% and their major fatty acid was anteiso-C(15:0). Strain CG1(T), selected as representative strain of the isolates, possesses meso-diaminopimelic acid in the cell wall peptidoglycan, MK-7 as the predominant menaquinone and diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine as the major polar lipids. Comparative 16S rRNA gene sequence analysis indicated that the isolates belonged to the genus Bacillus. The seven isolates shared 97.7-99.9% 16S rRNA gene sequence similarity, and formed a branch that was distinct from the type strains of the recognized species of the genus Bacillus. They were most closely related to Bacillus agaradhaerens DSM 8721(T) (92.6-93.8% 16S rRNA sequence similarity). DNA-DNA hybridization values between the seven isolates were 85-100%. According to the polyphasic characterization, the strains represent a novel species, for which the name Bacillus locisalis sp. nov. is proposed. The type strain is CG1(T) (CCM 7370(T)=CECT 7152(T)=CGMCC 1.6286(T)=DSM 18085(T)). PMID- 21640539 TI - Successful therapeutic hypothermia in patients with hypertrophic cardiomyopathy. PMID- 21640540 TI - Infected primary paraspinal hydatidosis with water lily sign. AB - BACKGROUND: Primary muscular hydatidosis is extremely rare, and the diagnosis of the infected muscular hydatid cyst can be difficult due to atypical clinical and radiological findings. CASE REPORT: We present herein an interesting case of a 24 year-old man with primary infected muscular hydatid cyst located in the paraspinal muscles eroding the right lamina of C6 vertebra that was diagnosed with water lily sign on computed tomography. CONCLUSION: Water lily sign is a pathognomonic imaging finding for hydatidosis, and defined when free-floating endocyst is seen. Cyst hydatid should always be considered in the differential diagnosis of any soft tissue mass of the patients from endemic regions. PMID- 21640541 TI - Sleep disorders screening, sleep health awareness, and patient follow-up by community pharmacists in Australia. AB - INTRODUCTION: Community pharmacies are well placed to deliver education and counseling to improve sleep health. OBJECTIVE: To develop, implement and evaluate a pharmacist led sleep health program. METHODS: Participating pharmacists (n=23) were trained to screen, counsel and follow up patients at risk of sleep disorders. Patients were screened for specific sleep disorders, counseled, provided written information and/or referred to physicians. Patient outcomes were evaluated by questionnaire at 3 or 12 months post screening. RESULTS: A total of 325 patients were screened, with 142 (44%) patients at risk of one or more sleep disorders. A total of 847 interventions were recorded by pharmacists: verbal counseling (49%), written information (34%), and referrals (16%). A total of 63 patients (of 99 recalling being referred) reported acting on their referral. Positive changes were made with smoking (4%), caffeine intake (10%), alcohol intake (9%) and improved sleep environment (19%). CONCLUSION: Pharmacists can raise awareness through educating patients on sleep health, and through counseling initiate behavior change in those at risk of having or developing a sleep disorder. PRACTICE IMPLICATIONS: Further research using this feasibility study can help in understanding the utility of pharmacists screening patients for sleep disorders and educating the public in sleep health. PMID- 21640542 TI - Analysis of pharmacist-patient communication using the Calgary-Cambridge guide. AB - OBJECTIVE: This study explored communication between pharmacists and patients through application of the Calgary-Cambridge guide [1] to appointment-based pharmacist-patient consultations and considers use of the guide in pharmacy education. METHODS: Eighteen patients attending appointment-based consultations with five pharmacists were recruited to this qualitative study. Consultations were audio-recorded and observed. Transcripts were coded according to the use of skills within the guide and analysed thematically. RESULTS: The results showed good use of many skills by pharmacists, particularly signposting and closing the session. Some skills were poorly represented such as listening effectively, eliciting the patient's perspective, effective use of computers and creating patient-centred consultations. A key theme of social conversation was present in the data but this skill was not defined in the guide. CONCLUSIONS: The Calgary Cambridge guide was developed for use in medical consultations but its application to pharmacist-patient consultations showed that the guide could be successfully used in pharmacy with some minor alterations. PRACTICE IMPLICATIONS: Pharmacists may need more training to improve the use of specific communication skills including how to conduct a patient-centred consultation. The Calgary Cambridge guide is well aligned with many aspects of pharmacist-patient consultations and could help pharmacists to improve their consultation skills. PMID- 21640543 TI - Patients' experiences and reported barriers to colonoscopy in the screening context--a systematic review of the literature. AB - OBJECTIVE: A systematic review of the literature was conducted to characterise patients' own experience of colonoscopy in the screening context. METHODS: A search strategy was applied in MEDLINE, EMBASE and PSYCHinfo (1996-2009). Thematic analysis and narrative summary techniques were used. RESULTS: Fifty-six studies met eligibility criteria and were included in the analysis. Seven studies examined patients' views after having colonoscopy. Forty-seven studies addressed patient-reported barriers to an anticipated primary colonoscopy. Most patients perceived the laxative bowel preparation to be the most burdensome part of colonoscopy. Other reported difficulties included anxiety, anticipation of pain, feelings of embarrassment and vulnerability. Inadequate knowledge and fear of finding cancer were identified as obstacles to the uptake of screening colonoscopy. Physician endorsement, having a family history, knowing someone with cancer, and perceived accuracy of the test were incentives to having a colonoscopy. Two studies focused on colonoscopy after faecal occult blood screening. Similar procedural, personal, and practical concerns were reported. CONCLUSIONS: Bowel preparation, lack of awareness of the importance of screening, and feelings of vulnerability in women are all significant barriers to screening colonoscopy. PRACTICE IMPLICATIONS: Patient reported obstacles and barriers to screening colonoscopy needs to be addressed to improve adherence. PMID- 21640544 TI - Factors that influence trajectories of change in frequency of substance use and quality of life among adolescents receiving a brief intervention. AB - This study aimed to identify factors influencing trajectories of change in two outcome domains, frequency of substance use and quality of life (QOL), among adolescents receiving a brief evidence-based intervention. Participants were 106 adolescents, aged 13 to 21 years, who met criteria for a substance use disorder. The adolescents received a five-session intervention and completed four assessments over 12 months. Based upon a theoretical and empirical review, five putative predictors were tested: gender, age, severity of conduct disorder, severity of depression, and peer substance involvement. Results of a parallel process latent growth curve model indicated that higher peer substance involvement and conduct severity predicted higher frequency of use at baseline, whereas higher peer substance involvement and depression severity predicted poorer QOL at baseline. Counter to predictions, higher depression severity predicted greater improvements in QOL following substance use treatment. The implications of baseline risk factors on adolescents' response to intervention are discussed. PMID- 21640545 TI - Prognostic effect of steatosis on hepatocellular carcinoma patients after liver resection. AB - AIMS: Overweight/obesity is currently a common health issue that may cause many diseases, even malignancies. The influence of steatosis on long-term results of surgical treatment for hepatocellular carcinoma (HCC) is not well known. The aim of this study is to analyze the results of hepatectomy for HCC patients with steatosis. METHODS: The study included 1048 patients who underwent hepatectomy for HCC from 1999 to 2005. The patients were divided into two groups; group A patients without steatosis (n = 693) and group B patients with steatosis (n = 355). The clinicopathological data and long-term survival were analyzed. RESULTS: Mean tumor size in group B patients was smaller than that in group A patients (4.61 +/- 3.40 vs. 5.91 +/- 4.36 cm, p < 0.01). Group B patients showed lower tumor differentiation grade, lower vascular invasion rate and better 5-year overall survival compared to group A patients (61.2% vs. 50.1%, p = 0.001). By multivariate analysis, steatosis was found to be associated with well differentiated, small-sized, and less alpha-fetoprotein productive tumors. When focusing on the tumors >5 cm in diameter, group B patients had better survival rate than group A patients (p = 0.041). Vascular invasion and steatosis were independent prognostic factors for the overall survival. CONCLUSION: HCC in steatotic liver was less aggressive than that in non-steatotic liver. HCC patients with steatosis have better surgical outcomes than those without steatosis. Vascular invasion and steatosis were independent prognostic factors for the overall survival if tumors were >5 cm in diameter. PMID- 21640546 TI - Canadian provincial trends in antihypertensive drug prescriptions between 1996 and 2006. AB - BACKGROUND: Little is known regarding potential differences in antihypertensive prescribing practices at a Canadian provincial level. Our objective was to determine provincial differences in the use of antihypertensive drug therapy in Canada. METHODS: Using longitudinal drug data (IMS CompuScript database; IMS Health Canada), we examined the increase in number of prescriptions dispensed for all antihypertensive agents for each province over an 11-year period (1996-2006). RESULTS: Over the 11-year study period, antihypertensive prescriptions increased by 106.2% for single-drug therapy (from 35.8% in Prince Edward Island and Newfoundland to 167.2% in British Columbia) and by 112.8% (from 22.0% in New Brunswick to 216.0% in Quebec) for combination-drug therapy. Among drug classifications, angiotensin receptor blockers had the largest increase for single-drug therapy and angiotensin-converting enzyme inhibitors-diuretics for combination-drug therapy. There were marked provincial differences in the increase in total antihypertensive therapy, ranging from British Columbia, with an increase of 262%, to Prince Edward Island and Newfoundland, where the increase was 134%. CONCLUSION: Large increases in antihypertensive prescriptions occurred in all provinces of Canada, but the provinces varied substantially in the increase in total and drug-specific classes of antihypertensive drugs. The basis for provincial differences in antihypertensive prescriptions remains unknown and is likely multifactorial but may relate in part to initial provincial variations in diagnosis, treatment, and control of hypertension, as well as individual provincial drug policies. PMID- 21640547 TI - Body composition, physical fitness, functional performance, quality of life, and fatigue benefits of exercise for prostate cancer patients: a systematic review. AB - CONTEXT: Prostate cancer patients, especially those on androgen deprivation therapy (ADT), experience many symptoms that make it difficult to maintain their independence and quality of life. Because ADT acts by means of reducing testosterone production, exercise may offset many of the ADT side effects and those of the cancer itself. OBJECTIVES: This systematic review of the literature evaluates whether exercise could reduce symptoms and improve quality of life for prostate cancer patients. METHODS: Using relevant databases and key words, 12 training studies were found meeting the inclusion and exclusion criteria. RESULTS: Grade A level evidence was observed for the benefits of exercise in improving muscular endurance, aerobic endurance, and overall quality of life, as well as reducing fatigue in prostate cancer patients. Grade B evidence also suggested that exercise may improve prostate cancer patients' muscle mass, muscular strength, functional performance (walking and sit to stand speed), as well as health-related, social and physical quality of life. These effects appeared greater for group-rather than home-based-exercise, especially if these programs included resistance training. CONCLUSION: It is recommended that most prostate cancer patients be encouraged to exercise regularly by their clinicians and significant others. Where possible, this exercise should be group-based and include some resistance training. Future research in this area should directly compare group- and home-based, as well as resistance, aerobic, and combined resistance and aerobic training to better elucidate the most effective forms of exercise for this population and what factors affect initiation and adherence to such programs. PMID- 21640548 TI - A Phase II dose titration study of thalidomide for cancer-associated anorexia. AB - CONTEXT: Sixty-five percent of people with advanced cancer suffers from loss of appetite. Several inflammatory cytokines appear to cause appetite loss in animal models. Thalidomide is an immunomodulatory drug that has been associated with improved appetite in those with HIV infections and cancer. OBJECTIVES: We completed a two-stage Phase II dose titration study of thalidomide, the primary purpose of which was to assess appetite response to thalidomide in cancer associated anorexia. METHODS: Individuals older than 18 years of age with active cancer, loss of appetite by numerical rating scale (NRS), life expectancy of at least four weeks, and Eastern Cooperative Oncology Group performance status of 0 3 were entered into the study. Pretreatment screening included medical history, neurologic examination, and symptoms by NRS and categorical scale (CAT). Patients received 50mg of thalidomide by mouth at bedtime for two weeks. Individuals who did not respond were dose escalated to 100mg at night for two weeks. Assessment of appetite, early satiety, fatigue, insomnia, night sweats, pain, and quality of life (QOL) occurred at two-week intervals. Toxicity also was assessed. The primary outcome was appetite response defined as a two-point reduction in the NRS or a one-point improvement in the CAT. RESULTS: Thirty-five patients entered the study; 33 completed 14 days of therapy and were analyzed for efficacy and toxicity. Sixty-four percent who completed at least two weeks of thalidomide had improved appetite. The CAT scores for appetite, insomnia, and QOL improved significantly. The 95% confidence intervals did not overlap. Five participants dropped out because of toxicity: two before two weeks and three later. CONCLUSION: Thalidomide reduced multiple symptoms commonly associated with cancer related anorexia and improved QOL. Our findings confirmed and validated a previously published single-arm trial. A recent randomized trial demonstrated greater benefits when thalidomide is used combined with other agents to treat cancer-associated anorexia and cachexia. Thalidomide helped cancer-associated anorexia in most patients. It also improved insomnia and QOL in advanced cancer. PMID- 21640549 TI - A psychometric evaluation of measures of spirituality validated in culturally diverse palliative care populations. AB - CONTEXT: Despite the need to accurately measure spiritual outcomes in diverse palliative care populations, little attention has been paid to the properties of the tools currently in use. OBJECTIVES: This systematic review aimed to appraise the psychometric properties, multifaith appropriateness, and completion time of spiritual outcome measures validated in multicultural advanced cancer, HIV, or palliative care populations. METHODS: Eight databases were searched to identify relevant validation and research studies. A comprehensive search strategy included search terms in three categories: palliative care, spirituality, and outcome measurement. Inclusion criteria were: validated in advanced cancer, HIV, or palliative care populations and in an ethnically diverse context. Included tools were evaluated with respect to psychometric properties (validity, reproducibility, responsiveness, and interpretability), multifaith appropriateness, and time to complete. RESULTS: A total of 191 articles were identified, yielding 85 tools. Twenty-six tools (representing four families of measures and five individual tools) met the inclusion criteria. Twenty-four tools demonstrated good content validity and 12 demonstrated adequate internal consistency. Only eight tools demonstrated adequate construct validity, usually because specific hypotheses were not stated and tested. Seven tools demonstrated adequate test-retest reliability; two tools showed adequate responsiveness, and two met the interpretability criterion. Data on the religious faith of the population of validation were available for 11 tools; of these, eight were tested in multifaith populations. CONCLUSION: Results suggest that, at present, the McGill Quality of Life Questionnaire, the Measuring the Quality of Life of Seriously Ill Patients Questionnaire, and the Palliative Outcome Scale are the most appropriate multidimensional measures containing spiritual items for use in multicultural palliative care populations. However, none of these measures score perfectly on all psychometric criteria, and their multifaith appropriateness requires further testing. PMID- 21640550 TI - Drivers of transfusion decision making and quality of the evidence in orthopedic surgery: a systematic review of the literature. AB - Reasons for variation in transfusion practice in orthopedic surgery are not well understood. This systematic review identified and appraised the quality of the literature in this area to assess the impact of factors associated with the use of allogeneic red blood cell (RBC) transfusion in orthopedic procedures. MEDLINE and EMBASE databases were searched for relevant English language publications. Articles containing a range of MeSH and text terms regarding "blood transfusion," "predictors," and "multiple logistic regression" were retrieved. Articles that focused on patients undergoing orthopedic procedures and that met prespecified inclusion criteria were appraised in terms of potential bias and the appropriateness of statistical approach. A total of 3641 citations were retrieved, and 29 met the inclusion criteria for the review. Articles reported on a range of orthopedic procedures including total hip arthroplasty; total knee arthroplasty, total shoulder arthroplasty, and spinal surgery. Most studies were conducted in the United States (n = 12) or Canada (n = 5). Study quality was moderate; 50% or more of the quality criteria were assessed in 15 articles. Particular areas of concern were the lack of prospective studies, lack of clarity in defining the time interval between risk factor assessment and transfusion outcome, and lack of model validation. A narrative synthesis found that 2 factors consistently influenced the use of RBC transfusion-decreased hemoglobin (n = 25) and increased patient age (n = 18). Increased surgical complexity (n = 12), low body weight (n = 9), presence of additional comorbidities (n = 9), and female sex (n = 7) were also important factors. The general quality of the studies in the field is weak. However, low hemoglobin and increasing age were consistently identified as independent risk factors for RBC transfusion in orthopedic practice. Additional or alternative analytical approaches are required to obtain a more comprehensive, holistic understanding of the decision to transfuse RBCs to patients undergoing orthopedic surgery. PMID- 21640551 TI - Precursors to numeracy in kindergartners with specific language impairment. AB - The present study investigated to what extent children with specific language impairment (SLI) differ in their early numeracy skills, when compared to normal language achieving (NLA) children. It was also explored which precursors were related to the early numeracy skills in both groups. Sixty-one children with SLI (6; 1 years) and 111 NLA children (6; 2 years) were assessed on general intelligence, working memory, naming speed, linguistic ability and early numeracy skills. The results showed lower scores for the children with SLI on logical operations and numeral representations but not on numeral estimations. The variance in numeral estimations was explained by children's intelligence and visual spatial memory. Phonological awareness and grammatical ability explained the variance in logical operations and numeral representations whereas naming speed turned out to be an additional factor in predicting these early numeracy skills of children with SLI. PMID- 21640552 TI - Handwriting capacity in children newly diagnosed with Attention Deficit Hyperactivity Disorder. AB - Preliminary evidence suggests that children with Attention Deficit Hyperactivity Disorder (ADHD) may exhibit handwriting difficulties. However, the exact nature of these difficulties and the extent to which they may relate to motor or behavioural difficulties remains unclear. The aim of this study was to describe handwriting capacity in children newly diagnosed with ADHD and identify predictors of performance. Forty medication-naive children with ADHD (mean age 8.1 years) were evaluated with the Evaluation Tool of Children's Handwriting Manuscript, the Movement Assessment Battery for Children (M-ABC), the Developmental Test of Visual Motor Integration (VMI) and the Conner Global Index. An important subset (85.0%) exhibited manual dexterity difficulties. Handwriting performance was extremely variable in terms of speed and legibility. VMI was the most important predictor of legibility. Upper extremity coordination, as measured by the M-ABC ball skills subtest, was also a good predictor of word legibility. CONCLUSION: Poor handwriting legibility and slow writing speed were common in children newly diagnosed with ADHD and were associated with motor abilities. Future studies are needed to determine whether interventions, including stimulant medications, can improve handwriting performance and related motor functioning. PMID- 21640553 TI - Recreational substance use patterns and co-morbid psychopathology in adults with intellectual disability. AB - There is very limited evidence on the patterns of recreational substance use among adults with Intellectual Disabilities (ID) who have co-morbid mental health problems. In this study we collected clinical and socio-demographic information as well as data on substance use patterns for consecutive new referrals (N = 115) to specialist mental health services for adults with ID in South-East London. The data were recorded from active clinical case notes. About 15% of patients had a history of substance use, however only 8% were currently using substances. Alcohol was the most frequently used substance (80%) followed by cannabis (28%) and cocaine (12%). Overall, substance use was significantly more likely among male patients, those with a mild level of ID and those with a forensic history. Substance use was less likely among patients with autism and more likely among those with schizophrenia spectrum disorders. Logistic regression analyses revealed that those with a forensic history were about five times more likely to have current substance use problems. Male gender was the only predictor for legal substance (alcohol) use. Illicit substance use was about three times more likely among patients with schizophrenia spectrum disorders. The present results highlight the role of illicit substance use as a health risk factor for adults with ID as well as the need to increase awareness within specialist mental health services. PMID- 21640554 TI - Simulation-based power calculation for designing interrupted time series analyses of health policy interventions. AB - OBJECTIVE: Interrupted time series is a strong quasi-experimental research design to evaluate the impacts of health policy interventions. Using simulation methods, we estimated the power requirements for interrupted time series studies under various scenarios. STUDY DESIGN AND SETTING: Simulations were conducted to estimate the power of segmented autoregressive (AR) error models when autocorrelation ranged from -0.9 to 0.9 and effect size was 0.5, 1.0, and 2.0, investigating balanced and unbalanced numbers of time periods before and after an intervention. Simple scenarios of autoregressive conditional heteroskedasticity (ARCH) models were also explored. RESULTS: For AR models, power increased when sample size or effect size increased, and tended to decrease when autocorrelation increased. Compared with a balanced number of study periods before and after an intervention, designs with unbalanced numbers of periods had less power, although that was not the case for ARCH models. CONCLUSION: The power to detect effect size 1.0 appeared to be reasonable for many practical applications with a moderate or large number of time points in the study equally divided around the intervention. Investigators should be cautious when the expected effect size is small or the number of time points is small. We recommend conducting various simulations before investigation. PMID- 21640555 TI - Measuring evidence-based medicine knowledge and skills. The Dutch Berlin Questionnaire: translation and validation. PMID- 21640556 TI - Rationale and design of the Japanese heart failure outpatients disease management and cardiac evaluation (J-HOMECARE). AB - BACKGROUND: Although many studies have demonstrated the efficacy of disease management programs on mortality, morbidity, quality of life (QOL), and medical cost in patients with heart failure (HF), no study has focused on psychological status as an outcome of disease management. In addition, very little information is available on the effectiveness of disease management programs in other areas than the USA and Europe. METHODS: The Japanese Heart Failure Outpatients Disease Management and Cardiac Evaluation (J-HOMECARE) is a randomized controlled trial in which 156 patients hospitalized with HF will be randomized into usual care or a home-based disease management arm receiving comprehensive advice and counseling by visiting nurses during the initial 2 months and telephone follow-up for the following 4 months after discharge. This study evaluates depression and anxiety (Hospital Anxiety and Depression Scale), mortality, readmission due to HF, and QOL (Short Form-8). Data are collected during index hospitalization and then 2, 6, and 12 months after discharge. This study started in December 2007, and the final results are expected in 2011. CONCLUSION: The J-HOMECARE will provide important information on the efficacy of disease management for psychological status as well as the effective components of disease management for patients with HF. (ClinicalTrials.gov number, NCT01284400). PMID- 21640558 TI - Detecting order-disorder transitions in discourse: implications for schizophrenia. AB - Several psychiatric and neurological conditions affect the semantic organization and content of a patient's speech. Specifically, the discourse of patients with schizophrenia is frequently characterized as lacking coherence. The evaluation of disturbances in discourse is often used in diagnosis and in assessing treatment efficacy, and is an important factor in prognosis. Measuring these deviations, such as "loss of meaning" and incoherence, is difficult and requires substantial human effort. Computational procedures can be employed to characterize the nature of the anomalies in discourse. We present a set of new tools derived from network theory and information science that may assist in empirical and clinical studies of communication patterns in patients, and provide the foundation for future automatic procedures. First we review information science and complex network approaches to measuring semantic coherence, and then we introduce a representation of discourse that allows for the computation of measures of disorganization. Finally we apply these tools to speech transcriptions from patients and a healthy participant, illustrating the implications and potential of this novel framework. PMID- 21640557 TI - Facial emotion recognition and facial affect display in schizotypal personality disorder. AB - BACKGROUND: Patients with schizophrenia have deficits in facial affect expression and detection that hinder social interactions. The goal of this study was to examine whether or not epidemiologically-related antipsychotic-naive schizotypal personality disorder (SPD) subjects would have similar deficits as patients with schizophrenia. METHODS: Matched SPD and healthy comparison (HC) subjects were asked to identify the eight classic emotions (SPD N=55, HC N=67) and to discriminate gender. Subjects (SPD N=22, HC N=17) were also photographed while displaying the same emotional expressions. Raters scored the subjects' facial expressions along several dimensions. RESULTS: SPD subjects compared with HC were slower and less accurate in identifying facial expressions. This may have been driven by deficits in identifying gender. Although raters were able to identify correctly SPD and HC subjects' expressions equally well, raters found SPD subjects' facial expressions to be more odd, more ambiguous, and the subjects less attractive in general compared with HC subjects. Raters were less confident in their ability to correctly interpret SPD subjects' facial expressions and raters were less comfortable with the idea of spending time with the SPD subjects compared with HC subjects. CONCLUSIONS: SPD subjects face two hurdles in terms of daily social interactions. They have problems both in correctly interpreting others' facial expressions and in generating socially attractive and unambiguous facial expressions. PMID- 21640559 TI - Antibiotic treatment for acute haematogenous osteomyelitis of childhood: moving towards shorter courses and oral administration. AB - Acute haematogenous osteomyelitis (AHOM) of childhood usually affects the long bones of the lower limbs. Although almost any agent may cause AHOM, Staphylococcus aureus is the most common bacterium, followed by Streptococcus pneumoniae and, in some countries, Salmonella spp. and Kingella kingae. Magnetic resonance imaging (MRI) has improved the diagnostic accuracy of traditional radiography and scintigraphy. Except for the pre-treatment diagnostic sample from bone before the institution of antibiotic therapy, no other surgery is usually required. Traditionally, non-neonatal AHOM has been treated with a 1-3-month course of antibiotics, including an intravenous (i.v.) phase for the first weeks, but recent prospective randomised studies challenge this approach. For most uncomplicated cases, a course of 20 days including an i.v. period of 2-4 days suffices, provided large enough doses of a well-absorbed agent (clindamycin or a first-generation cephalosporin, local resistance permitting) are used, administration is four times daily and most symptoms and signs subside within a few days. Serum C-reactive protein (CRP) is a good guide in monitoring the course of illness, and the antimicrobial can usually be discontinued if CRP has decreased to <20 mg/L. Newer and costly agents, such as linezolid, should be reserved for cases due to resistant S. aureus strains. AHOM in neonates and immunocompromised patients probably requires a different approach. Because sequelae may develop slowly, follow-up for at least 1 year post hospitalisation is recommended. PMID- 21640560 TI - Setting interpretive breakpoints for antimicrobial susceptibility testing using disk diffusion. AB - Antimicrobial susceptibility testing plays a key role in clinical microbiology. The disk diffusion test dates back to the 1940s and became standardised from the 1950s, with the International Collaborative Study (ICS) and National Committee for Clinical Laboratory Standards (NCCLS) as the two major standards. Interlaboratory variation of disk test results was recognised early but has never been dealt with in a satisfactory manner. The error-rate bounded method was described in 1974 and its role is discussed. Species-specific susceptibility interpretation was coined in 1980 for Proteus mirabilis and chloramphenicol. In the late 1970s, more extensive use of species-specific breakpoints was introduced in Lund (Sweden). At the same time, P. Mouton constructed species-specific regression lines and pointed out the difficulties with narrow ranges of minimal inhibitory concentration (MIC) values. A more general use of species-specific regression lines was made possible with single-strain regression analysis, using one well-defined strain tested in disk diffusion with a range of disk contents. This method made it possible to calibrate the disk test in an individual laboratory. Other methods to achieve such calibration are also described. A recent method, 'MIC-coloured zone diameter histogram-technique', has proven useful for the validation of species-specific interpretive breakpoints. The microbiological breakpoint proposed by Williams in 1990 has experienced a renaissance with the European Committee on Antimicrobial Susceptibility Testing (EUCAST) epidemiological cut-off value (ECOFF). MIC and zone diameter distributions with accompanying ECOFFs for species-antimicrobial combinations are published on the EUCAST website. A method for the reconstruction of wild-type zone diameter populations, namely normalised resistance interpretation, is described. This method can produce resistance figures that are truly comparable between laboratories. PMID- 21640561 TI - Quantitative investigation, stability and in vitro release studies of anti-TB drugs in Triton niosomes. AB - The highly stable innocuous niosomes composed of four components (Triton X 100, polyethylene glycol 2000, water, Span 80) have been prepared successfully and characterized using particle size analyzer, transmission and scanning electron microscopy. The mean size has been found to be in the range 200-300 nm. The optimization of niosomes has been carried out using fluorescence spectroscopy. An attempt has been made to incorporate anti-tuberculosis drugs (ATD's) in the prepared niosomes. The stability and encapsulation efficiency of these drugs in the niosome have also been assessed and high encapsulation efficiency is observed. Such high encapsulation efficiency will serve as an advantage to solve the problem of multi-drug resistance in case of tuberculosis. Release studies and kinetics have been carried out to investigate the release behavior of drugs from the prepared niosomes. Fickian or diffusional release has been observed for rifampicin and isoniazid and a non-Fickian release mechanism for pyrazinamide. Fluorescence probe quenching technique has been used to determine the location and distribution coefficient of the ATD's in niosome/water system. PMID- 21640562 TI - Studies on interaction of colloidal silver nanoparticles (SNPs) with five different bacterial species. AB - Silver nanoparticles (SNPs) are being increasingly used in many consumer products like textile fabrics, cosmetics, washing machines, food and drug products owing to its excellent antimicrobial properties. Here we have studied the adsorption and toxicity of SNPs on bacterial species such as Pseudomonas aeruginosa, Micrococcus luteus, Bacillus subtilis, Bacillus barbaricus and Klebsiella pneumoniae. The influence of zeta potential on the adsorption of SNPs on bacterial cell surface was investigated at acidic, neutral and alkaline pH and with varying salt (NaCl) concentrations (0.05, 0.1, 0.5, 1 and 1.5 M). The survival rate of bacterial species decreased with increase in adsorption of SNPs. Maximum adsorption and toxicity was observed at pH 5, and NaCl concentration of <0.5 M. A very less adsorption was observed at pH 9 and NaCl concentration >0.5 M, there by resulting in less toxicity. The zeta potential study suggests that, the adsorption of SNPs on the cell surface was related to electrostatic force of attraction. The equilibrium and kinetics of the adsorption process were also studied. The adsorption equilibrium isotherms fitted well to the Langmuir model. The kinetics of adsorption fitted best to pseudo-first-order. These findings form a basis for interpreting the interaction of nanoparticles with environmental bacterial species. PMID- 21640563 TI - Facile green synthesis of gold nanoparticles using leaf extract of antidiabetic potent Cassia auriculata. AB - A simple biological method for the synthesis of gold nanoparticles (AuNPs) using Cassia auriculata aqueous leaf extract has been carried out in the present study. The reduction of auric chloride led to the formation of AuNPs within 10 min at room temperature (28 degrees C), suggesting a higher reaction rate than chemical methods involved in the synthesis. The size, shape and elemental analysis were carried out using X-ray diffraction, TEM, SEM-EDAX, FT-IR and visible absorption spectroscopy. Stable, triangular and spherical crystalline AuNPs with well defined dimensions of average size of 15-25 nm were synthesized using C. auriculata. Effect of pH was also studied to check the stability of AuNPs. The main aim of the investigation is to synthesize AuNPs using antidiabetic potent medicinal plant. The stabilizing and reducing molecules of nanoparticles may promote anti-hyperglycemic if tested further. PMID- 21640564 TI - Bacterial adhesion on hybrid cationic nanoparticle-polymer brush surfaces: ionic strength tunes capture from monovalent to multivalent binding. AB - This paper describes the creation of hybrid surfaces containing cationic nanoparticles and biocompatible PEG (polyethylene glycol) brushes that manipulate bacterial adhesion for potential diagnostic and implant applications. Here, ~10 nm cationically functionalized gold nanoparticles are immobilized randomly on negative silica surfaces at tightly controlled surface loadings, and the remaining areas are functionalized with a hydrated PEG brush, using a graft copolymer of poly-l-lysine and PEG (PLL-PEG), containing 2000 molecular weight PEG chains and roughly 30% functionalization of the PLL. The cationic nanoparticles attract the negative surfaces of suspended Staphylococcus aureus bacteria while the PEG brush exerts a steric repulsion. With the nanoparticle and PEG brush heights on the same lengthscale, variations in ionic strength are demonstrated to profoundly influence the capture of S. aureus on these surfaces. For bacteria captured from gentle flow, a crossover from multivalent to univalent binding is demonstrated as the Debye length is increased from 1 to 4 nm. In the univalent regime, 1 um diameter spherical bacteria are captured and held by single nanoparticles. In the multivalent regime, there is an adhesion threshold in the surface density of nanoparticles needed for bacterial capture. The paper also documents an interesting effect concerning the relaxations in the PLL-PEG brush itself. For brushy surfaces containing no nanoparticles, bacterial adhesion persists on newly formed brushes, but is nearly eliminated after these brushes relax, at constant mass in buffer for 12h. Thus brushy relaxations increase biocompatibility. PMID- 21640565 TI - Characterisation of exosomes derived from human cells by nanoparticle tracking analysis and scanning electron microscopy. AB - Exosomes from three different cell types (HEK 293T, ECFC, MSC) were characterised by scanning electron microscopy (SEM), dynamic light scattering (DLS) and nanoparticle tracking analysis (NTA). The diameter was around 110 nm for the three cell types. The stability of exosomes was examined during storage at -20 degrees C, 4 degrees C, and 37 degrees C. The size of the exosomes decreased at 4 degrees C and 37 degrees C, indicating a structural change or degradation. Multiple freezing to -20 degrees C and thawing did not affect the exosome size. Multiple ultracentrifugation also did not change the exosome size. PMID- 21640566 TI - Synthesis and characterization of nano-composite ion-exchanger; its adsorption behavior. AB - A novel organic-inorganic nanocomposite cation-exchanger has been synthesized via sol-gel method. It was characterized on the basis of FTIR, XRD, SEM, TEM, AFM and Raman studies. The structural studies reveal semi-crystalline nature of the material with the particle size ranging from 1-5 nm. Physiochemical properties such as ion-exchange capacity, chemical and thermal stability of composite material have also been determined. Bifunctional behavior of the material has been indicated by its pH titrations curves. The nanocomposite material exhibits improved thermal stability, higher ion-exchange capacity and better selectivity for toxic heavy metals. The ion-exchange material shows an ion-exchange capacity of 1.8 meq g(-1) for Na(+) ions. Sorption behavior of metal ions on the material was studied in different solvents. The cation exchanger was found to be selective for Pb(II), Hg(II) and Zr(IV) ions. The limit of detection (LOD) and the limit of quantification (LOQ) for Pb(II) ion was found to be 0.85 and 2.85 MUg L(-1). Analytically important separations of heavy metal ions in synthetic mixtures as well as industrial effluents and natural water were achieved with the exchanger. The practical utility of polyanilineZr(IV)sulphosalicylate cation exchanger has been established for the analysis and recovery of heavy metal ions in environmental samples. PMID- 21640567 TI - Oral administration of caffeic acid ameliorates the effect of cisplatin on brush border membrane enzymes and antioxidant system in rat intestine. AB - Cisplatin (CP) is a widely used antineoplastic drug that exhibits gastrointestinal toxicity. We have previously shown that administration of a single dose of CP results in a decrease in the activities of several brush border membrane (BBM) enzymes, induces oxidative stress and alters the activities of several antioxidant enzymes in the small intestine of rats. In the present study we have investigated the effect of treatment with the dietary antioxidant caffeic acid (CA) on CP induced biochemical changes in the intestine. Administration of a single intraperitoneal dose of CP alone (6 mg/kg body weight) led to a decrease in the activities of the BBM enzymes, increase in lipid peroxidation, decrease in sulfhydryl groups and changes in the activities of catalase, superoxide dismutase, glutathione peroxidase, glucose 6-phosphate dehydrogenase, glutathione reductase, glutathione S-transferase and thioredoxin reductase. Administration of two doses of CA (each of 250 mg/kg body weight), at 15 and 120 min after treatment with CP, significantly attenuated the CP-induced changes in all these parameters but the administration of CA alone had no effect. These results suggest that CA is an effective agent in reducing the effects of CP on the intestine and could prove to be useful in alleviating the gastrointestinal toxicity of this drug. PMID- 21640568 TI - Regulation of cardiac oxidative stress and lipid peroxidation in streptozotocin induced diabetic rats treated with aqueous extract of Pimpinella tirupatiensis tuberous root. AB - Plants with antidiabetic activities provide important source for the development of new drugs in the management of diabetes mellitus. The main aim of this study was to evaluate the protective effect of aqueous extract (AE) of Pimpinella tirupatiensis (Pt) tuberous root on cardiac oxidative stress and lipid peroxidation (LPO) in non-diabetic and streptozotocin (STZ)-induced diabetic rats. Diabetes was induced in male Wistar rats by a single administration of STZ (40 mg/kg intraperitoneal (i.p). AE (750 mg/kg/b.w./day) and glibenclamide (GLB) (20 mg/kg/b.w./day) were administrated orally by intra oral gastric tube for 30 days. After 4 weeks of hyperglycaemia the enzymatic and non-enzymatic factors were measured in cardiac tissue of diabetic and control groups. Xanthine oxidase activity (XOD), Uric acid (UA) and malondialdehyde (MDA) content were significantly (p<0.01) elevated by 48, 48 and 50% respectively and the contents of glutathione (GSH), ascorbic acid (AA) were significantly (p<0.01) diminished by 45 and 42% respectively in diabetic rats when compared to normal. Treatment with AE and GLB normalized the content of UA, GSH, AA, MDA and the activity of XOD. No significant changes were observed in control rats treated with AE. This data suggests that hyperglycemia induces oxidative stress in the heart, but the oxidative stress defense mechanisms in the heart tissue are fairly efficacious against oxidative injury by the treatment with AE and GLB. The present study reveals that AE may provide a useful therapeutic option in the reversal of oxidative stress induced cardiac dysfunction in diabetes mellitus. PMID- 21640569 TI - Mylodon darwinii DNA sequences from ancient fecal hair shafts. AB - Preserved hair has been increasingly used as an ancient DNA source in high throughput sequencing endeavors, and it may actually offer several advantages compared to more traditional ancient DNA substrates like bone. However, cold environments have yielded the most informative ancient hair specimens, while its preservation, and thus utility, in temperate regions is not well documented. Coprolites could represent a previously underutilized preservation substrate for hairs, which, if present therein, represent macroscopic packages of specific cells that are relatively simple to separate, clean and process. In this pilot study, we report amplicons 147-152 base pairs in length (w/primers) from hair shafts preserved in a south Chilean coprolite attributed to Darwin's extinct ground sloth, Mylodon darwinii. Our results suggest that hairs preserved in coprolites from temperate cave environments can serve as an effective source of ancient DNA. This bodes well for potential molecular-based population and phylogeographic studies on sloths, several species of which have been understudied despite leaving numerous coprolites in caves across of the Americas. PMID- 21640570 TI - Outcome measures of peripheral nerve regeneration. AB - Animal models of nerve compression, crush, and transection injuries of peripheral nerves have been subject to extensive study in order to understand the mechanisms of injury and axon regeneration and to investigate methods to promote axon regeneration and improve functional outcomes following nerve injury. Six outcome measures of regenerative success including axon and neuron counts, muscle and motor unit contractile forces, and behavior are reviewed in the context of nerve injury types, crush, transection and nerve repair by direct coaptation, or transection and repair via a nerve graft or conduit. The measures are evaluated for sciatic, tibial, common peroneal, femoral, single nerve branches such as the soleus nerve, and facial nerves. Their validity is discussed in the context of study objectives and the nerve branch. The case is made that outcome measures of axon counts and muscle contractile forces may be valid during the early phases of axon regeneration when regenerating sprouts emerge asynchronously from the proximal nerve stump and regenerate towards their denervated targets. However, care must be taken especially when experimental interventions differentially affect how many neurons regenerate axons and the number of axons per neuron that sprout from the proximal nerve stumps. Examples of erroneous conclusions are given to illustrate the need for researchers to ensure that the appropriate outcome measures are used in the evaluation of the success of peripheral nerve regeneration. PMID- 21640571 TI - Maxillary sinus floor augmentation using a nano-crystalline hydroxyapatite silica gel: case series and 3-month preliminary histological results. AB - The aim of this case series is to histologically examine a new hydroxyapatite in sinus lift procedure after 3 months. Ten 2-stage sinus lifts were performed in 10 healthy patients having initial bone height of 1-2mm and bone width of 5mm, asking for a fixed implant-supported rehabilitation. After graft material augmentation, a rough-surfaced mini-implant was inserted to maintain stability of the sinus widow. A bioptical core containing a mini-implant was retrieved 3 months after maxillary sinus augmentation with NanoBone((r)) and processed for undecalcified histology. From the histomorphometric analysis, NanoBone((r)) residuals accounted for the 38.26% +/- 8.07% of the bioptical volume, marrow spaces for the 29.23% +/- 5.18% and bone for the 32.51% +/- 4.96% (new bone: 20.64% +/- 2.96%, native bone: 11.87% +/- 3.27%). Well-mineralized regenerated bone with lamellar parallel-fibred structure and Haversian systems surrounded the residual NanoBone((r)) particles. The measured bone-to-implant contact amounted to 26.02% +/- 5.46%. No connective tissue was observed at the implant boundary surface. In conclusion, the tested material showed good histological outcomes also 3 months after surgery. In such critical conditions, the use of a rough surfaced mini-implant showed BIC values supposed to be effective also in case of functional loading. Although longer follow-up and a wider patient size are needed, these preliminary results encourage further research on this biomaterial for implant load also under early stage and critical conditions. PMID- 21640572 TI - Simple, rapid and sensitive detection of antibiotics based on the side-by-side assembly of gold nanorod probes. AB - In this work, we report on the application of versatile gold nanorods (GNRs) in optical sensors for the detection of antibiotics. The target analyte, Gentamicin (GM) and ovalbumin (OVA)-antigen-modified GNRs together competed with antibody modified GNRs, then influenced the formation of side-by-side aggregates of the GNRs by antibody-antigen interactions. Accordingly, the UV-vis absorption intensity of the side-by-side aggregates was changed in the presence of the target analyte. This assay allowed the selective determination of GM in the range of 0.1-20 ng/mL, and the limit of detection (LOD) of GM was 0.05 ng/mL. Furthermore, compared with the traditional plate-based immunoassay, the developed method was easy to perform without washing cycles and the results could be read as soon as the nanoprobe-analyte incubation was complete. Therefore, the developed method could be a promising tool for the detection of antibiotic residues. PMID- 21640573 TI - Osteosarcoma: A rare cause of painful enlargement of the hallux. AB - Malignant osseous and soft-tissue tumors of the foot are rare. We report a case of osteosarcoma in the proximal phalanx of the hallux in a 45-year-old man. In patients with foot-related symptoms, a high index of suspicion for pedal osteosarcoma is required. Delayed or inappropriate diagnosis may compromise limb sparing surgery and survivorship. PMID- 21640574 TI - Postoperative analgesia with tramadol and indomethacin for diagnostic curettage and early termination of pregnancy. AB - BACKGROUND: The postoperative analgesic effects of rectal indomethacin and tramadol were compared in patients undergoing elective termination of first trimester pregnancy and diagnostic dilatation and curettage. METHODS: Eighty-one American Society of Anesthesiologists class I and II women undergoing first trimester termination of pregnancy or diagnostic dilation and curettage were randomly allocated to receive rectal suppositories of either tramadol 100 mg (n=41) or indomethacin 100 mg (n=40) 90 min before induction of anesthesia. Pain scores and side effects were evaluated until discharge. Intraoperative anesthetic and postoperative analgesic consumption was also recorded. Intravenous metamizole 1 g was employed for postoperative rescue analgesia. RESULTS: When compared to the indomethacin group, the tramadol group required less intraoperative propofol [136 mg +/-28 vs. 160 mg +/-35 (P=0.001)], less rescue analgesia [2.4% vs. 22% (P=0.005)] and lower visual analogue pain scores [2.4 +/-8 vs. 23 +/-22 (P=0.005)]. The incidence of postoperative nausea and vomiting was similar in both groups. CONCLUSION: When compared to indomethacin 100 mg, preoperative administration of tramadol 100 mg provides superior postoperative analgesia with minimal adverse effects. PMID- 21640575 TI - Skin disinfection before spinal anaesthesia for caesarean section. PMID- 21640576 TI - Is a relatively high pre-spinal heart rate associated with reduced efficacy of prophylactic vasopressor during spinal anaesthesia for caesarean section? PMID- 21640577 TI - Non-invasive measurement of hemoglobin during cesarean hysterectomy: a case series. AB - Obstetric patients diagnosed with abnormal placentation (placenta accreta, increta or percreta) are at increased risk of major postpartum hemorrhage and cesarean hysterectomy. Obstetric anesthesiologists are primarily involved in intraoperative transfusion management in these cases. Hemoglobin assessment is invaluable for assisting transfusion decision-making during the acute period of obstetric hemorrhage. However, laboratory and point-of-care tests of hemoglobin concentration are time-dependent and intermittent, and do not provide a real-time assessment of change during the acute phase of blood loss. A new non-invasive hemoglobin monitor has been introduced recently, which provides real-time measurement of hemoglobin values (SpHb) using multi-wavelength pulse co-oximetry. We present a review of five patients with suspected abnormal placentation who received SpHb monitoring during cesarean hysterectomy at our institution. We discuss the potential clinical utility of non-invasive hemoglobin monitoring for pregnant patients at high risk of obstetric hemorrhage, and the potential role of SpHb in guiding transfusion therapy. PMID- 21640578 TI - Intravenous mannitol for treatment of abducens nerve paralysis after spinal anesthesia. PMID- 21640579 TI - Prolonged neurological deficit following neuraxial blockade for caesarean section. PMID- 21640580 TI - Labour analgesia and the baby: good news is no news. PMID- 21640581 TI - Proximal vascular control in cases of abnormal placentation. PMID- 21640582 TI - Supercritical water gasification of biomass: Thermodynamic constraints. AB - In the present work, the supercritical water gasification (SCWG) of biomass is analyzed with a view to outlining the possible thermodynamic constraints that must be taken into account to develop this new process. In particular, issues concerning the formation of solid carbon and the process heat duty are discussed. The analysis is conducted by means of a two-phase non-stoichiometric thermodynamic model, based on Gibbs free energy minimization. Results show that char formation at equilibrium only occurs at high biomass concentrations, with a strong dependence on biomass composition. As regards the process heat duty, SCWG is mostly endothermic when biomass concentration is low, although a very small amount of oxidizing agent is able to make the process exothermic, with only a small loss in the heating value of the syngas produced. PMID- 21640583 TI - Ethanol production from marine algal hydrolysates using Escherichia coli KO11. AB - Algae biomass is a potential raw material for the production of biofuels and other chemicals. In this study, biomass of the marine algae, Ulva lactuca, Gelidium amansii,Laminaria japonica, and Sargassum fulvellum, was treated with acid and commercially available hydrolytic enzymes. The hydrolysates contained glucose, mannose, galactose, and mannitol, among other sugars, at different ratios. The Laminaria japonica hydrolysate contained up to 30.5% mannitol and 6.98% glucose in the hydrolysate solids. Ethanogenic recombinant Escherichia coli KO11 was able to utilize both mannitol and glucose and produced 0.4g ethanol per g of carbohydrate when cultured in L. japonica hydrolysate supplemented with Luria-Bertani medium and hydrolytic enzymes. The strategy of acid hydrolysis followed by simultaneous enzyme treatment and inoculation with E. coli KO11 could be a viable strategy to produce ethanol from marine alga biomass. PMID- 21640584 TI - Sensitive luciferin derived probes for selective carboxypeptidase activity. AB - Highly selective luminescent probes, QLUC-TYR and LUC-GLU, for detection of carboxypeptidase activity were synthesized. Caged substrates were first cleaved by corresponding carboxypeptidases, and then they were activated by luciferase to emit light. Enzymatic activities of biologically important carboxypeptidases can be determined using this technology. PMID- 21640585 TI - RETRACTED: Sannastatin, a novel toxic macrolactam polyketide glycoside produced by actinomycete Streptomyces sannanensis. AB - This article has been retracted: please see Elsevier Policy on Article Withdrawal (http://www.elsevier.com/locate/withdrawalpolicy).This article has been retracted at the request of the Editor-in-Chief. One of the co-authors, Hartmut Laatsch, informed the journal that this paper was submitted and published without his consent. This is a violation of the journal's policy that all authors of a paper should approve submission of a manuscript. Apologies are offered to readers of the journal that this was not detected during the submission and evaluation process. PMID- 21640586 TI - A new route to dTDP-6-deoxy-l-talose and dTDP-L-rhamnose: dTDP-L-rhamnose 4 epimerase in Burkholderia thailandensis. AB - dTDP-L-rhamnose (dTDP-Rha)-synthesizing dTDP-6-deoxy-L-lyxo-4-hexulose reductase (4-KR) and dTDP-Rha 4-epimerase were characterized from Burkholderia thailandensis E264 by utilizing rmlD(Bth) (BTH_I1472) and wbiB(Bth) (BTH_I1476), respectively. Incubation of the recombinant WbiB(Bth) with RmlA/RmlB/RmlC/Tal, which has previously been shown to generate dTDP-6-deoxy-L-talose (dTDP-6dTal) from alpha-D-glucose-1-phosphate, dTTP, and NADPH, produced dTDP-Rha. (1)H NMR measurements confirmed that both RmlA/RmlB/RmlC/Tal/WbiB(Bth) and RmlA/RmlB/RmlC/RmlD produced dTDP-Rha. WbiB(Bth) alone produced dTDP-Rha when incubated with dTDP-6dTal. This is the first report to demonstrate epimerase activity interconverting between dTDP-Rha and dTDP-6dTal. PMID- 21640587 TI - Synthesis of novel pyrazolo[1,5-a]pyrazin-4(5H)-one derivatives and their inhibition against growth of A549 and H322 lung cancer cells. AB - A series of substituted pyrazolo[1,5-a]pyrazin-4(5H)-one was synthesized by the reaction of ethyl 1-(2-oxo-2-phenylethyl)-3-phenyl-1H-pyrazole-5-carboxylate derivatives and 2-(2-aminoethoxy)ethanol or 2-morpholinoethanamine in the condition of microwave-assisted one-step and solvent-free in a good yield. The structures of the compounds were determined by IR, (1)H NMR and mass spectroscopy. In addition, a representative single-crystal structure was characterized by using X-ray diffraction analysis. Preliminary biological evaluation showed that the compounds could inhibit the growth of A549 and H322 cells in dosage-dependent manners. PMID- 21640588 TI - Design, synthesis and activity of a potent, selective series of N-aryl pyridinone inhibitors of p38 kinase. AB - A series of N-aryl pyridinone inhibitors of p38 mitogen activated protein (MAP) kinase were designed and prepared based on the screening hit SC-25028 (1) and structural comparisons to VX-745 (5). The focus of the investigation targeted the dependence of potency and metabolic stability on the benzyloxy connectivity, the role of the C-6 position and the substitution pattern on the N-phenyl ring. Further optimization produced the highly selective and potent pyridinones 2 and 3. These inhibitors exhibited activity in both acute and chronic models of inflammation. PMID- 21640589 TI - The molecular mechanisms of interactions between bioactive peptides and angiotensin-converting enzyme. AB - The ability of milk protein derived Ile-Pro-Ala (IPA), Phe-Pro (FP) and Gly-Lys Pro (GKP) peptides to inhibit angiotensin I-converting enzyme (ACE), a protein with an important role in blood-pressure regulation, were verified in vitro and in vivo. This work elucidates the modes and molecular mechanisms of the interaction of IPA, FP and GKP with ACE, including mechanisms that bind the peptides to the cofactor Zn(2+). It was observed that the best docking poses obtained for IPA, FP and GKP were at the ACE catalytic site with very similar modes of interaction, including the interaction with Zn(2+). The interactions, including H-bonds, hydrophobic, hydrophilic, and electrostatic interactions, as well as the interaction with Zn(2+), were responsible for the binding between the bioactive peptides and ACE. PMID- 21640590 TI - Intraductal papillomas of the breast: diagnosis and management of 151 patients. AB - INTRODUCTION: The assessment of papillary lesions continues to be a challenging area in breast radiology and pathology. The management of intraductal papillomas without atypia of the breast remains controversial. The purpose of the present study was to determine diagnostic accuracy of radiographical diagnosis, core biopsy, and surgical excision in papillary breast lesions. MATERIAL AND METHODS: By using files from 1995 to 2010, 151 cases of intraductal papilloma with or without atypia were identified. Patients were stratified as follows: core biopsy followed by surgical excision (n = 61), core biopsy alone (n = 19), and surgical excision alone (n = 71). RESULTS: The upstage rate of intraductal papillomas without atypia on core biopsy to atypia or malignancy on excision was 8.9%. Excision specimens revealed intraductal papillomas without atypia in 68 out of 71 cases, and atypical papillomas in 3 cases. CONCLUSION: Our findings suggest that radiographic and histopathological diagnosis of intraductal papillomas show high accuracy and good concordance. In cases where the radiographic diagnosis reveals suspicious lesions core biopsy represents the first choice. PMID- 21640591 TI - The influence of Nordic Walking training on sit-to-stand transfer in Parkinson patients. AB - Neurodegenerative processes in Parkinson's disease (PD) particularly affect activities of daily living (ADL). Problems of patients with PD in sit-to-stand (STS) performance have been verified before, but not the effects of training on biomechanical measures of STS function. This pilot study aimed to analyse effects of 12 weeks of Nordic Walking training and severity of PD: healthy controls (CO), least (UPDRS A) and more severe (UPDRS B) affected PA on selected functional outcome measures. We expected improvements in PD similar to CO, with better performance of the unstable second phase and faster execution of the entire movement with higher velocities of centre of gravity (COG). 3D kinematics of 22 PD and 18 CO subjects before and after training, were recorded using a motion analysis system (Vicon, Oxford). We compared five outcome measures for STS in 11 PD and 11 CO, matched according to age, gender, height, and weight. Effects of Nordic Walking training were not statistically significant but indicated different patterns which depended on the values of patient's UPDRS score (part III, motor functions). Time required for STS performance increased and horizontal and vertical velocity of COG decreased in UPDRS B, which could be due to progression of PD during the training period. In contrast, UPDRS A showed similar effects as CO. The effects of Nordic Walking as an easy, economic and low-risk intervention on STS in PD depend on the degree of PD. Our findings may help scientists, patients, and therapists to adjust sport-physiological interventions. PMID- 21640592 TI - Ste20-related kinases: effectors of signaling and morphogenesis in fungi. AB - The family of Ste20-related kinases is conserved from yeast to mammals and includes the p21 activated kinases (PAKs) and germinal centre kinases (GCKs). These kinases have been shown to be involved in signaling through mitogen activated protein kinase (MAPK) pathways and in morphogenesis through the regulation of cytokinesis and actin-dependent polarized growth. This review concentrates on the role of Ste20-related kinases in fungi where recent research has revealed roles for both PAKs and GCKs in the regulation of cytokinesis and in previously unidentified roles in promoting hyphal growth and differentiation of asexual development structures. In particular, the importance of PAKs during pathogenesis will be examined. PMID- 21640593 TI - Exploratory analysis of kinetic solubility measurements of a small molecule library. AB - Kinetic solubility measurements using prototypical assay buffer conditions are presented for a ~58,000 member library of small molecules. Analyses of the data based upon physical and calculated properties of each individual molecule were performed and resulting trends were considered in the context of commonly held opinions of how physicochemical properties influence aqueous solubility. We further analyze the data using a decision tree model for solubility prediction and via a multi-dimensional assessment of physicochemical relationships to solubility in the context of specific 'rule-breakers' relative to common dogma. The role of solubility as a determinant of assay outcome is also considered based upon each compound's cross-assay activity score for a collection of publicly available screening results. Further, the role of solubility as a governing factor for colloidal aggregation formation within a specified assay setting is examined and considered as a possible cause of a high cross-assay activity score. The results of this solubility profile should aid chemists during library design and optimization efforts and represent a useful training set for computational solubility prediction. PMID- 21640594 TI - New class azaphilone produced by a marine fish-derived Chaetomium globosum. The stereochemistry and biological activities. AB - Four new metabolites, chaetomugilins P-R and 11-epi-chaetomugilin I, were isolated from a strain of Chaetomium globosum originally obtained from the marine fish Mugil cephalus, and their absolute stereostructures were elucidated on the basis of spectroscopic analyses, including 1D and 2D NMR techniques and various chemical transformations. Particularly, the skeleton of chaetomugilin P is different from that of other azaphilones isolated from this fungal strain to date. In addition, these compounds significantly inhibited the growth of cultured P388, HL-60, L1210 and KB cell lines. PMID- 21640595 TI - Plastic scintillators with beta-diketone Eu complexes for high ionizing radiation detection. AB - Luminescent and scintillation properties of polystyrene-based plastic scintillators with beta-diketone Eu complexes are investigated. A scintillator with dibenzoylmethane Eu complex containing two phenyl groups demonstrates the maximum scintillating efficiency. It is shown that plastic scintillators efficiency is dramatically decreased if beta-diketone derivatives contain no phenyl groups as substituents. This fact can be explained by exciplex mechanism of energy transfer from a matrix to Eu complex. PMID- 21640596 TI - Implementation and characterisation of new neutron imaging system for dynamic processes investigation at the Es-Salam research reactor. AB - Neutron imaging is a powerful method for non-destructive investigations where high penetration through metals and in particular high contrast for hydrogenous materials maybe exploited. Due to the complexity of digital neutron static or video image formation, image capture conditions and parameters must be accurately selected. In this work, implementation of a new neutron imaging system based on CCD camera and LiF-ZnS scintillator is presented. The image characteristics in terms of contrast, noise and dynamic range and investigation limits of this new imaging system were studied as a function of the neutron source properties. PMID- 21640598 TI - How to do clinical research in cranio-maxillo-facial surgery. AB - INTRODUCTION: Not many randomised controlled trials are published in surgical journals, especially those on maxillo-facial surgery. There appears to be some uncertainty on how to perform such studies. Accordingly this paper offers some information on how to plan, pursue and publish a well conducted case-control study, or the more powerful randomised control trial. RESULT: The main section describes how to define a relevant clinical question, and a research protocol, the way to implement the study, and it helps to find funding for such research. It also explains the various study designs, gives a very short introduction to statistics and on how to appraise the results achieved, and it advises on writing and submitting the resultant manuscript. CONCLUSION: This paper offers a guide for young colleagues who wish to perform a study, write a paper and achieve publication in one of our leading speciality journals. PMID- 21640597 TI - Custom made bioceramic implants in complex and large cranial reconstruction: a two-year follow-up. AB - INTRODUCTION: Large cranial defects still represent a challenge in neurosurgery. Currently different biomaterials are available for cranial reconstruction including titanium, acrylic mesh and different types of calcium phosphate-based bone grafts. The goal of surgery is a perfect fit of the implant without infection and absorption, and a good aesthetic result. This paper describes a surgical method for cranioplasty, using a customised porous hydroxyapatite (HA) prosthesis. MATERIALS AND METHODS: Sixty patients treated surgically with a customised porous-HA prosthesis for large cranial defects, were followed retrospectively. A two-year follow-up was carried out with periodic visits and CT scans. Safety (the incidence of adverse events and fractures of the implant) and clinical performance (biological and cosmetic results) were evaluated. RESULTS: Fifty one patients were followed-up, no rejection occurred and only one case of infection was recorded. Five patients had minor surgery-related complications, and no spontaneous implant fractures or mobilisation were reported. Three patients exhibited implant fractures as a result of trauma and all healed spontaneously. All patients showed a satisfactory clinical outcome with good cosmetic appearance in the early postoperative period and after a long-term follow-up. CONCLUSIONS: Cranioplasty performed with a customised porous-HA prosthesis gave a positive outcome, showing it to be an appropriate technique for use in large and complex cranial reconstruction. PMID- 21640599 TI - Total subapical mandibular osteotomy to correct class 2 division 1 dento-facial deformity. AB - A severe Class 2 division 1 dento-facial deformity in an adult is normally treated by a combination of orthodontics and orthognathic surgery. The surgical procedure may involve either a single or double jaw osteotomy. The sagittal split osteotomy with forward mandibular slide has been the procedure of choice for many years for correcting the antero-posterior discrepancy between the dental arches. Although the total subapical mandibular osteotomy was described over 20 years ago, reports of its use in the literature are sparse. We report two cases to demonstrate that it can be the operation of choice for surgical correction of Class 2 dento-facial deformity in carefully selected cases. When performed correctly it can have very good aesthetic and functional results. PMID- 21640600 TI - Details in a meta-analysis comparing mitral valve repair to replacement for ischemic regurgitation. PMID- 21640601 TI - Mechanical circulatory support after heart transplantation. AB - OBJECTIVE: Mechanical circulatory support (MCS) may be used for severe graft failure after heart transplantation, but the degree to which it is lifesaving is uncertain. METHODS: Between June 1990 and December 2009, 53 patients after 1417 heart transplants (3.7%) required post-transplant MCS for acute rejection (n=17), biventricular failure (n=16), right ventricular failure (n=16), left ventricular failure (n=1), or respiratory failure (n=3). Although support was occasionally instituted remotely post-transplant (5>1 year), in 39 (73%) instances it was required within 1 week. Initial mode of support was extracorporeal membrane oxygenation in 43 patients (81%), biventricular assist device in 4 (7.5%), and right ventricular assist device in 6 (11%). RESULTS: Risk of requiring respiratory support was highest in those with restrictive cardiomyopathy as indication for transplant, women, and those with elevated pulmonary pressure or renal failure. Complications of support, which increased progressively with its duration, included stroke in two patients (3.8%), infection in two (3.8%), and reoperation for bleeding (seven instances) in four (7.0%). Nineteen patients (36%) recovered and were removed from support, five (9.4%) underwent retransplantation (four after biventricular failure and one after acute rejection), and 29 died while on support (55%). Overall survival after initiating support was 94%, 83%, 66%, and 43% at 1, 3, 7, and 30 days, respectively. Patients requiring support for biventricular failure had better survival than those having acute rejection or other indications (P=0.03). Survival after retransplantation or removal from support following recovery was 88% at 1 year and 61% at 10 years. CONCLUSION: Severe refractory heart failure after transplantation is a rare catastrophic event for which MCS offers the possibility of recovery or bridge to retransplantation, particularly for patients with biventricular failure in the absence of rejection. Early retransplantation should be considered in patients who show no evidence of graft recovery on MCS. PMID- 21640602 TI - Estimating the risk of complications related to re-exploration for bleeding after adult cardiac surgery: a systematic review and meta-analysis. AB - OBJECTIVE: The aim of this study was to evaluate the impact of re-exploration for bleeding after cardiac surgery on the immediate postoperative outcome. METHODS: Systematic review of the literature and meta-analysis of data on re-exploration for bleeding after adult cardiac surgery were performed. RESULTS: The literature search yielded eight observational studies reporting on 557,923 patients and were included in the present analysis. Patients requiring re-exploration were significantly older, more frequently males, had a higher prevalence of peripheral vascular disease and preoperative exposure to aspirin, and more frequently underwent urgent/emergency surgery. Re-exploration was associated with significantly increased risk ratio (RR) of immediate postoperative mortality (RR 3.27, 95% confidence interval (CI) 2.44-4.37), stroke, need of intra-aortic balloon pump, acute renal failure, sternal wound infection, and prolonged mechanical ventilation. The pooled analysis of four studies (two being propensity score-matched pairs analysis) reporting adjusted risk for mortality led to an RR of 2.56 (95%CI 1.46-4.50). Studies published during the last decade tended to report a higher risk of re-exploration-related mortality (RR 4.30, 95%CI 3.09 5.97) than those published in the 1990s (RR 2.75, 95%CI 2.06-3.66). CONCLUSIONS: This study suggests that re-exploration for bleeding after cardiac surgery carries a significantly increased risk of postoperative mortality and morbidity. PMID- 21640604 TI - Cardiac echinococcosis: surviving 40 years later? PMID- 21640605 TI - G-CSF therapy and catheter-related Gram-positive sepsis increase serum IL-2 receptor alpha level and may falsely suggest a relapse in children with soft tissue sarcomas unless serum beta2-microglobulin, lactate dehydrogenase and C reactive protein levels are determined concomitantly. AB - Many components of oncologic treatment increase serum sIL-2Ralpha level, which may falsely suggest a relapse. We tried to establish whether granulocyte colony stimulating factor (G-CSF) and central vein catheter (CVC)-related sepsis increase serum sIL-2Ralpha level to values on relapse of childhood soft tissue sarcomas (STS) and how to distinguish real relapse from a "false" one. Serum sIL 2Ralpha, B2-M, LDH, CRP and ESR levels and rates of markers' elevated values were determined prospectively in 18 STS children: pre-treatmently (ST1), in complete remission (CR; ST2), in CR during G-CSF therapy (ST3), in CR during CVC-related sepsis (ST4), on relapse (ST5) and after treatment (ST6) and once in 50 healthy pediatric controls. It appeared that pre-treatment serum sIL-2Ralpha, LDH, CRP and ESR but not B2-M declined significantly with remission (ST2) achievement. At ST5 sIL-2Ralpha, B2-M, LDH and CRP increased from ST2 to ST1 values. SIL-2Ralpha levels at ST3 and ST4 rose significantly in all patients from ST2 to ST1 and ST5 values. At ST3 also serum LDH and B2-M increased to values at ST1 and ST5 and exceeded significantly those at ST2 and ST4. At ST4 CRP but not B2-M and LDH, rose significantly in most patients to values at ST1 and ST5. Thus, serum sIL 2Ralpha monitoring in pediatric STS reflects well response to chemotherapy unless samples are collected during G-CSF therapy or CVC-related sepsis. Determination of serum B2-M, LDH and CRP together with sIL-2Ralpha may help to distinguish between "real" relapse and "false" sIL-2Ralpha increase due to G-CSF administration or CVC-related sepsis. PMID- 21640606 TI - How can we tell who is aware? Where does the veracity lie? AB - BACKGROUND: The first aim of this study was to compare the subjective experiences of social, cognitive, and emotional problems of the patient and his/her next of kin, and explore if this related to cognitive testing. The second aim was to explore how these results reflect the patient's awareness. METHODS: This is a subanalysis from a longitudinal study in the first year after discharge with comparisons of patient and next of kin scores on the European Brain Injury Questionnaire (EBIQ) and analyses of the relationship of their scores to an objective cognitive screening on 3 occasions. A paired t test was used to explore differences between 35 stroke patients and their next of kin on the EBIQ. Gamma analyses were made to explore the relationship between the EBIQ scores and the Barrow Neurological Institute Screening (BNIS) of higher cognitive functions, with the entire sample grouped into aware and unaware according to the BNIS item of awareness. RESULTS: We found significant differences between the patient and next of kin assessments on all occasions, apart from at discharge. When grouped, only the group of aware patients differed significantly from their next of kin. Significant relations of the patient ratings on the EBIQ and BNIS were only found at the 1-year follow-up for the unaware group and the entire sample. Next of kin EBIQ scores did not correlate with the BNIS. CONCLUSIONS: Neither next of kin ratings nor an objective measurement was feasible to use for evaluation of the patient's awareness of social, cognitive, and emotional problems. PMID- 21640607 TI - Positive caregiving experiences are associated with life satisfaction in spouses of stroke survivors. AB - BACKGROUND: Studies into caregivers usually have been focused on negative caregiving experiences. This study is based on the hypotheses that positive caregiving experiences (i.e., self-esteem derived from caregiving) of spouses of stroke patients also need to be taken into account, and that these are related to life satisfaction in 2 ways: first, by a direct association with life satisfaction, and second, indirectly by way of a buffer effect (i.e., by compensating for the impact of negative caregiving experiences on life satisfaction). METHODS: In this cross-sectional study (n = 121) 3 years poststroke, the Caregiver Reaction Assessment was used to assess caregiver burden (Burden) and self-esteem derived from caregiving (Self-esteem scale). Life satisfaction was measured with the Life Satisfaction Questionnaire (LiSat-9). Spearman correlations and regression analyses were performed. RESULTS: Both Self esteem and Burden scores were associated with life satisfaction (correlation coefficients 0.35 and -0.74, respectively). An interaction effect was also found (P = .006); spouses who perceived both high Burden and high Self-esteem reported significantly higher life satisfaction scores (mean 4.2, standard deviation [SD] 0.5) than spouses who perceived high Burden but low Self-esteem (mean 3.6, SD 0.7). CONCLUSIONS: Positive caregiving experiences are related to spouses' life satisfaction 3 years poststroke and mediate the impact of burden on life satisfaction. Positive caregiving experiences should get more attention in rehabilitation research and practice. PMID- 21640608 TI - Barriers to the use of intravenous tissue plasminogen activator for in-hospital strokes. AB - BACKGROUND: Patients who have ischemic strokes (ISs) while hospitalized for other conditions may be less likely to receive intravenous tissue plasminogen activator (IV tPA) when compared to patients who have strokes in the community. This study explored possible barriers to IV tPA use in these patients. METHODS: Stroke diagnosis was confirmed by chart review for all adult patients admitted to Bellevue Hospital between January 1, 2004 and December 31, 2008 who were discharged with a primary or secondary International Classification of Diseases, 9th edition code for transient ischemic attack, intracerebral hemorrhage, or IS. Circumstances at stroke onset were recorded for all patients who had strokes while hospitalized for another reason. RESULTS: Seventy-nine in-hospital IS cases were identified; 18 (23%) occurred <2 weeks after major surgery, and another 14 (18%) had a delayed diagnosis because signs were not readily detectable on clinical examination. Twenty-four patients (30%) were eligible for IV tPA, of whom 13 were identified within 3 hours of onset and 10 (13%) were treated with IV tPA. The median National Institutes of Health Stroke Scale score was higher in hospitalized patients (13) than in patients admitted through the emergency department (5; P < .001 using the Wilcoxon rank sum test). CONCLUSIONS: Seventy percent of in-hospital IS cases in our single hospital retrospective study were postoperative, clinically subtle, or had contraindications to IV tPA, preventing its use. Of the remaining untreated patients, the biggest barrier to IV tPA administration was delay in stroke discovery, which was largely dependent on observation by hospital staff or family rather than patient report. PMID- 21640609 TI - A model to prevent fibrinolysis in patients with stroke mimics. AB - BACKGROUND: Many patients with stroke-mimicking conditions receive treatment with intravenous fibrinolysis (IVF), a treatment associated with potentially serious complications. We sought to determine if any clinical or radiographic characteristics can help predict stroke mimics among IVF candidates. METHODS: This retrospective study was carried out at a single institution. Patients treated with intravenous recombinant tissue plasminogen activator (rt-PA; n = 193) were divided into 3 categories: acute ischemic stroke (n = 142), aborted stroke (n = 21), and stroke mimics (n = 30). Analysis of variance and the chi square test were used to assess differences, while logistic regression models were computed to predict groups. RESULTS: Mimics treated with rt-PA did not experience complications (intracranial bleeding, systemic hemorrhage, or angioedema), and had better neurologic and functional outcomes than stroke patients (P < .05). Several variables helped differentiate strokes from mimics, including atherosclerosis on computed tomographic angiography (odds ratio [OR] 23.6; 95% confidence interval [CI] 8.4-66.2), atrial fibrillation (OR 11.4; 95% CI 1.5-86.3), age >50 years (OR 7.2; 95% CI 2.8-18.5), and focal weakness (OR 4.15; 95% CI 1.75-9.8). Other variables decreased chances of stroke: migraine history (OR 0.05; 95% CI 0.01-0.4), epilepsy (OR 0.13; 95% CI 0.02-0.8), paresthesia (OR 0.1; 95% CI 0.04-0.3), and precordialgia (OR 0.045; 95% CI 0.002 0.9). A regression model using focal weakness, computed tomographic angiography findings, and precordialgia had a 90.2% predictive accuracy. CONCLUSIONS: IVF has low complication rates in stroke mimics. Certain clinical characteristics appear predictive of stroke mimics, particularly normal computed tomographic angiography. If confirmed, this may help prevent giving IVF to patients without stroke. PMID- 21640610 TI - Safety of periprocedural heparin in acute ischemic stroke endovascular therapy: the multi MERCI trial. AB - BACKGROUND: There are limited data on the safety of periprocedural heparin in acute ischemic stroke endovascular therapy. METHODS: A post hoc analysis was performed on patients enrolled in the Multi Mechanical Embolus Removal in Cerebral Ischemia (MERCI) trial to compare baseline characteristics and clinical outcomes between patients who received periprocedural heparin (HEP(+)) with patients who did not receive periprocedural heparin (HEP(-)). Data on periprocedural heparin use or nonuse was collected on patients enrolled between February 1, 2006 and July 31, 2006. RESULTS: Of 51 patients included in the analysis cohort, 24 (47%) received periprocedural heparin with a median dose of 3000 U. Baseline and procedural characteristics were similar between the 2 groups, although HEP(+) patients were more likely to have vertebral or basilar occlusion than HEP(-) patients (16.7% v 0%; P = .04). There was no significant difference in rates of hemorrhage, procedural complications, or 90-day mortality between the 2 groups. In multivariable analysis, a 90-day good outcome (modified Rankin scale score of 0-2) was associated with age (odds ratio [OR] 0.92; 95% confidence interval [CI] 0.86-0.98; P = .0104), final revascularization success (OR 6.86; 95% CI 1.39-33.81; P = .0179), and periprocedural heparin use (OR 5.89; 95% CI 1.34-25.92; P = .0189). CONCLUSIONS: In this small subgroup of the Multi MERCI trial, periprocedural heparin use in acute ischemic stroke endovascular therapy was not associated with increased rates of intracerebral hemorrhage or 90 day mortality. The improved 90-day good outcome among patients undergoing mechanical thrombectomy combined with periprocedural heparin warrants further study in a larger cohort. PMID- 21640611 TI - Pilot study of cardiac magnetic resonance imaging for detection of embolic source after ischemic stroke. AB - BACKGROUND: Transesophageal echocardiography (TEE) is the standard for evaluating cardioembolic sources of stroke, although many strokes remain cryptogenic after TEE. Cardiac magnetic resonance (CMR) imaging may have advantages over TEE. We performed a prospective pilot study comparing CMR to TEE after stroke to assist in planning future definitive studies. METHODS: Individuals with nonlacunar stroke within 90 days of undergoing clinical TEE were prospectively identified and underwent a 1.5 Tesla research CMR scan. Exclusion criteria included >50% relevant cervical vessel stenosis and inability to undergo nonsedated CMR. A descriptive comparison of cardioembolic source (intracardiac thrombus/mass, aortic atheroma >= 4 mm, or patent foramen ovale [PFO]) by study type was performed. RESULTS: Twenty patients underwent CMR and TEE a median of 6 days apart. The median age was 51 years (interquartile range [IQR] 40, 63.5), 40% had hypertension, 15% had diabetes, 25% had a previous stroke/transient ischemic attack, 5% had atrial fibrillation, and none had coronary disease or heart failure. No patient had intracardiac thrombus or mass detected on either study. Aortic atheroma >= 4 mm thick was identified by TEE in 1 patient. CMR identified aortic atheroma as <4 mm in this patient (3 mm on CMR compared with 5 mm on TEE). PFO was identified in 6 of 20 patients on TEE; CMR found only 1 of these. CONCLUSIONS: In this pilot study, TEE identified more potential cardioembolic sources than CMR imaging. Future studies comparing TEE and CMR after stroke should focus on older subjects at higher risk for cardiac disease to determine whether TEE, CMR, or both can best elucidate potential cardioembolic sources. PMID- 21640612 TI - Gradient echo T2*-weighted magnetic resonance imaging revealing cerebral microbleeds in a patient with microscopic polyangiitis complicated by cerebrovascular disease. AB - A 67-year-old woman developed alveolar hemorrhage and was positive for the myeloperoxidase antineutrophil cytoplasmic antibody (MPO-ANCA). She was diagnosed as having probable microscopic polyangiitis (MPA) in accordance with the Japanese criteria. Brain computed tomographic and magnetic resonance imaging (MRI) scans revealed asymptomatic chronic cerebral hemorrhage and white matter lesions. She also later developed thalamic infarction. Gradient echo T2-weighted MRI showed cerebral microbleeds. Cerebral microbleeds in our patient represent bleeding prone necrotizing vasculitis, which is a characteristic pathologic feature of MPA. Although cerebrovascular disease is not major complication of MPA, cerebral hemorrhage occurs more frequently than ischemic infarction, and it can be critical and fatal. Brain MRIs, including gradient echo T2-weighted imaging, should be performed on patients diagnosed as or suspected of having MPA to assess the risk of cerebral hemorrhage. PMID- 21640613 TI - Management of tight intraoperative glycemic control during off-pump coronary artery bypass surgery in diabetic and nondiabetic patients. AB - OBJECTIVES: To optimize intra- and postoperative insulin management in cardiac surgical patients. DESIGN: A prospective, randomized, open-label, single-center study. SETTING: A large nonuniversity hospital. PARTICIPANTS: Sixty diabetics and 60 nondiabetics undergoing off-pump cardiac bypass surgery. INTERVENTIONS: Intra- and postoperative tight glycemic control were achieved using different approaches with a modified insulin protocol. MEASUREMENTS AND MAIN RESULTS: Nondiabetics were divided randomly: in the ND-ind group (n = 30), insulin was started at induction according to preinduction blood glucose (BG) concentrations. In group ND >110 (n = 30), insulin was started when BG concentrations exceeded 110 mg/dL during surgery. Up to 85% of the ND >110 group started on insulin intraoperatively. Intraoperatively, the ND-ind group had more BG within target (80-110 mg/dL) (p = 0.002), less BG >130 mg/dL (p = 0.015), and more BG between 70 and 79 mg/dL (p = 0.002). In diabetics, BG concentration was checked every 30 (DM-30), n = 30) versus 60 minutes (DM-60, n = 30) to improve the protocol's performance. Intraoperatively, there were more BG concentrations within target (80-110 mg/dL) (p = 0.02) and less >130 mg/dL (p = 0.0002) in the DM-30 group. During surgery, the hyperglycemic index and the glycemic penalty index were lower in the ND-ind group (p < 0.05). Postoperatively, the mean BG concentrations, hyperglycemic index, and glycemic penalty index in diabetics and nondiabetics were comparable between groups (p < 0.05). In the overall 2,641 BG samples, the lowest BG concentration in the operating room was 71 and in the intensive care unit (ICU) it was 61 mg/dL. CONCLUSIONS: In diabetics and nondiabetics undergoing off-pump coronary artery bypass surgery, tight perioperative glycemic control is feasible and efficient, with minimal risks for hypo- and hyperglycemia. In nondiabetics, starting insulin therapy from induction onwards results in more measurements within target, without affecting the mean BG. In diabetics, decreasing the sampling interval from 60 to 30 minutes results in more measurements within target and in a mean blood glucose within target at ICU arrival. PMID- 21640614 TI - Should an anesthesiologist be present on site during cardiologic interventions? PMID- 21640615 TI - Use of an inferior epigastic arterial catheter in a patient with difficult vascular access. PMID- 21640616 TI - A multimodal approach to address the inflammation of cardiopulmonary bypass? PMID- 21640617 TI - Diverse morphologic manifestations of cardiac allograft vasculopathy: a pathologic study of 64 allograft hearts. AB - BACKGROUND: Cardiac allograft vasculopathy (CAV) is a major limitation to the long-term success of cardiac transplantation. Although there are published descriptions of the lesions, there have been no studies delineating the pathology of CAV in a large series of patients who underwent retransplantation for CAV. METHODS: We reviewed archival records and microscopic sections of surgically explanted hearts from 64 patients who underwent cardiac retransplantation: 54 adults (18 to 70 years old) and 10 children (3 to 15 years old). Vascular lesions were categorized as showing intimal fibromuscular hyperplasia, atherosclerosis and/or inflammation. The degree of luminal narrowing was estimated from gross descriptions and microscopic sections. RESULTS: In total, 75% of hearts had evidence of acute cellular rejection, mostly mild. Intramyocardial arteries showed primarily intimal fibromuscular hyperplasia and inflammation with no atheromas present. Large and branch epicardial coronary arteries were narrowed in at least one artery of all hearts. Lesions in the epicardial coronary arteries were composed of intimal fibromuscular hyperplasia, atherosclerosis and/or inflammation affecting one or more vascular layers (intima, media and adventitia). Severe CAV with >75% luminal narrowing was seen in the LAD in 17% of hearts, the LCx in 17% and the RCA in 22% of hearts. Two hearts had severe narrowing of the left main coronary artery. Nineteen arteries had luminal thrombi. All hearts had narrowing of smaller epicardial branch coronary arteries that was often severe. Atheromas were present in arteries of adults and children; thus, not all atheromas could be considered pre-existing prior to transplantation. Both arteries and veins showed intimal hyperplasia and inflammation. CONCLUSIONS: CAV is a pathologically multifaceted disorder that affects large and small epicardial coronary arteries of adults and children, with different types of lesions: intimal fibromuscular hyperplasia; atherosclerosis; and/or inflammation (vasculitis). Therapies to address this disease must take into account the protean nature of the vascular lesions. PMID- 21640618 TI - Successful use of a pneumatic biventricular assist device as a bridge to transplantation in cardiogenic shock. AB - BACKGROUND: Mechanical circulatory support is a highly effective technology to maintain organ perfusion in patients with cardiogenic shock as a bridge to transplantation. Although implantation of a left ventricular assist device alone is often the preferred configuration, patients with biventricular failure and significant end-organ dysfunction often require biventricular assistance. METHODS: Between January 2000 and September 2008, 80 patients with severe biventricular failure were accepted for heart transplantation and received a pneumatic biventricular assist devices as a bridge to transplant. Patients were retrospectively divided into 2 groups: those successfully bridged to transplant (Group A) and those who died (Group B). Patients were also divided into 2 periods of implantation: Group X (2000-2005) and Group Y (2006-2008, which used a multidiscipline selection process). RESULTS: Overall success rate to transplantation was 71.3%, with Group Y demonstrating an 82% success to transplant rate vs 63% in Group X. One-year actuarial survival after transplant was 89% compared with 92% in patients without a ventricular assist device. There were no statistically significant laboratory parameters between Groups A and B identifying potential risk factors for poor outcome. CONCLUSION: Biventricular assist device therapy represents an effective and reliable means of supporting selected Interagency Registry for Mechanically Assisted Circulatory Support profile 1 patients as a bridge to transplantation, with excellent success to transplant rates and post-transplant survival. PMID- 21640620 TI - A novel myoepithelial cell marker in canine mammary tissue. PMID- 21640619 TI - Focal positive prostate-specific membrane antigen (PSMA) expression in ganglionic tissues associated with prostate neurovascular bundle: implications for novel intraoperative PSMA-based fluorescent imaging techniques. AB - OBJECTIVE: Prostate specific membrane antigen (PSMA) is primarily expressed in glandular prostatic tissue and is frequently utilized to detect primary or metastatic prostatic adenocarcinoma (CaP). A purported novel application of PSMA detection is the intraoperative real-time identification of CaP using radioimmunoscintigraphy to define the extension of the surgical resection. Considering that PSMA expression has been reported in other tissues, we evaluated its immunoexpression in prostatic neurovascular bundle elements to assess the convenience and safety of the aforementioned procedure. MATERIALS AND METHODS: Twenty consecutive specimens of radical prostatectomy (RP) were retrieved from our surgical pathology archives. PSMA immunoexpression (Clone 3E6, DAKO) was assessed in a representative section from each specimen containing neurovascular bundle elements. RESULTS: PSMA expression was documented in 20/20 of examined CaP slides. Most cases exhibited an apical/cytoplasmic or cytoplasmic with membranous accentuation pattern of staining. Focal weak to moderate cytoplasmic staining was detected in associated ganglionic tissue in 3/15 of the examined RP. In all cases, staining was cytoplasmic, less extensive, and weaker than the pattern observed in CaP. None of the peripheral nerve sheath cells or lymphovascular components of the examined neurovascular bundles were positive for PSMA. CONCLUSIONS: We found focal positive PSMA expression in the ganglionic cells of the prostatic neurovascular bundle. Our results suggest that the radioimmunoscintigraphic detection of radiolabeled PSMA antibodies might not be entirely specific for prostatic cells; this observation must be taken into account should an intraoperative PSMA-based fluorescent imaging technique be used to define the extension of the surgical procedure. PMID- 21640621 TI - Cardiac arrest and post resuscitation of the brain. AB - Primary out-of-hospital cardiac arrest in childhood is rare but survival is a little better for children than for adults, although the prognosis for infants is very poor. Hypoxic-ischaemic encephalopathy after in-hospital cardiac arrest in children undergoing complicated treatment for previously untreatable conditions is now a common problem and is probably increasing. An additional ischaemic insult worsens the prognosis for other encephalopathies, such as that occurring after accidental or non-accidental head injury. For near-drowning, the prognosis is often good, provided that cardiopulmonary resuscitation (CPR) is commenced immediately, and the child gasps within 40 minutes of rescue and regains consciousness soon afterwards. The prognosis is much worse for the nearly drowned child admitted to casualty or the emergency room deeply unconscious with fixed dilated pupils, requiring continuing CPR and with an arterial pH <7, especially if there is little recovery by the time of admission to the intensive care unit. The use of adrenaline, sodium bicarbonate and calcium appears to worsen prognosis. Neurophysiology, specifically serial electroencephalography and evoked potentials, is the most useful tool prognostically, although neuroimaging and biomarkers may play a role. In a series of 89 patients studied after cardiac arrest in three London centres between 1982 and 1985, 39% recovered consciousness within one month. Twenty seven percent died a cardiac death whilst in coma, and the outcome in the remainder was either brain death or vegetative state. EEG and initial pH were the best predictors of outcome in this study. Seizures affected one third and were associated with deterioration and worse outcome. The advent of extracorporeal membrane oxygenation (ECMO) and the positive results of hypothermia trials in neonates and adults have rekindled interest in timely management of this important group of patients. PMID- 21640622 TI - Estrogen receptors localization in the spinal trigeminal nucleus: an immunohistochemical study in humans. AB - There is increasing evidence for estrogenic modulation of neurotransmission within the trigeminal pain pathway. It is also likely that the effects of estrogens may be influenced by the presence and localization of estrogen receptors (ERs) in a given brain area. To date, human data on the localization of ERs in the spinal trigeminal nucleus (STN), a key brain region in craniofacial nociception, are lacking. To ascertain whether ERs are expressed in the human STN, we performed immunohistochemical analysis on medulla oblongata samples taken from eight adult subjects (three men and five women; age range, 23-71 years) who had died from causes unrelated to neurologic or endocrine diseases. Paraffin embedded sections at the level of the subnucleus caudalis and interpolaris were incubated with anti-estrogen receptor alpha (ERalpha) and anti-estrogen receptor beta (ERbeta) antibodies. ERalpha immunoreactivity was detected in the nucleus and cytoplasm of neuronal and glial cells in the STN and in the nerve fibers within the spinal trigeminal tract in all eight subjects; ERbeta immunoreactivity was observed in the cytoplasm of neuronal cells in five subjects. This study is the first to provide evidence in humans that ER immunoreactivity is detectable on neuronal and glial cells of the STN. The two ER subtypes exhibited different expression patterns, with higher expression levels of ERalpha than ERbeta. The presence of ER-containing cells in the STN suggests that estrogens may directly affect trigeminal neuron excitability in humans. PMID- 21640623 TI - Mechanisms regulating superoxide generation in experimental models of phenylketonuria: an essential role of NADPH oxidase. AB - This study was designed to investigate whether nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, a superoxide-producing enzyme, could be involved in phenylketonuria (PKU)-associated oxidative stress. A Pah(enu2)-BTBR PKU mouse model, and an in vitro cell culture model of PKU mimicking high phenylalanine insults in PKU, were employed for this study. The concentration of phenylalanine in mouse cerebral cortex was determined by liquid chromatography-tandem mass spectrometry. Superoxide production was displayed with dihydroethidium staining. NADPH oxidase expression level was measured by real-time RT-PCR, Western blotting and immunofluorescence. NADPH oxidase activity was measured by the colorimetric method. The phenylalanine concentrations in cerebral cortices of PKU mice were significantly higher than those in wild-type control mice. Similar results concerning superoxide production and NADPH oxidase protein expression and activity, were also found in this brain region. In addition, it was found that cerebral cortical neurons subjected to an in vitro high phenylalanine insult, displayed increased superoxide production accompanied by increases of NADPH oxidase protein expression and activity. Pretreatment with the inhibitor of this oxidase (diphenylene iodonium or apocynin) prevented this superoxide-increasing effect. Collectively, these findings provide evidence that NADPH oxidase might be a key enzyme involved in enhanced superoxide production in PKU and suggest that it may be a potential therapeutic target in neuroprotective strategies against phenylalanine-evoked oxidative brain injury in PKU. PMID- 21640624 TI - Effect of aerobic and resistance exercise training on late-onset Pompe disease patients receiving enzyme replacement therapy. AB - Pompe disease is a rare autosomal recessive disorder characterized by the deficiency of acid alpha-glycosidase resulting in lysosomal accumulation of glycogen. The late-onset disease form is characterized by progressive skeletal and respiratory muscle dysfunction. In addition to the recently introduced enzyme replacement therapy (ERT), treatments such as protein-enriched diet and exercise training have been proposed, although little is known about their effectiveness on the physical condition of such patients. Aim of the present study was to investigate the effect of exercise training on muscular strength and body composition in five patients with late-onset Pompe disease receiving ERT. All subjects followed a 20 week lasting program of supervised aerobic and progressive resistance exercise training. Before and after the training period, body composition was determined with dual X-ray absorptiometry and isometric muscular strength was measured with a specialized load transducer. Functional capacity was assessed using the 6-min shuttle walk test. A significant increase in muscular strength (15-50% at various body parts, p<0.05) and 6-minute walking distance (203.8 +/- 177 m before vs. 248.2 +/- 184 m after, p<0.01) was observed after training, whereas total and lower extremities lean body mass did not change significantly. These results suggest that exercise training has a positive effect on muscular strength and functional capacity in patients on ERT with late-onset Pompe disease. PMID- 21640625 TI - Presumptive meningoencephalitis secondary to extension of otitis media/interna caused by Streptococcus equi subspecies zooepidemicus in a cat. AB - A 5-year-old castrated male domestic longhair cat was presented with neurological signs consistent with a central vestibular lesion and left Horner's syndrome. Computed tomography images revealed hyperattenuating, moderately contrast enhancing material within the left tympanic bulla, most consistent with left otitis media/interna. Marked neutrophilic pleocytosis was identified on cerebrospinal fluid analysis. Streptococcus equi subspecies zooepidemicus (SEZ) was isolated from the cerebrospinal fluid. Intracranial extension of otitis media/interna is relatively infrequent in small animals. There are no reports of otitis media/interna caused by SEZ in dogs or cats. This is the first report of otitis media/interna and presumptive secondary meningoencephalitis caused by SEZ in a cat. PMID- 21640626 TI - Subclinical airway inflammation despite high-dose oral corticosteroid therapy in cats with lower airway disease. AB - Management of feline chronic lower airway disease focuses on controlling clinical signs and decreasing airway inflammation. This retrospective study evaluated the correlation between the resolution of clinical signs in cats with lower airway disease receiving oral glucocorticoids with the resolution of inflammation based on bronchoalveolar lavage fluid (BALF) cytology. Ten cats diagnosed with lower airway disease based on characteristic clinical signs and inflammatory BALF cytology received oral glucocorticoids for at least 3 weeks. They were required to have resolution of clinical signs and BALF collected while asymptomatic and still receiving glucocorticoids. Cats received prednisolone or prednisone (average dose of 1.8+/-0.2mg/kg daily) for 35.7+/-5.5 days. Three cats had resolution of clinical signs and lacked inflammatory BALF cytology; seven had persistent inflammatory BALF cytology despite resolution of clinical signs. Given that subclinical inflammation during high-dose glucocorticoid treatment was common, current recommendations to taper therapy based on resolution of clinical signs should be re-evaluated. PMID- 21640627 TI - [Evaluation of the postoperative acute pain after heart surgery]. AB - INTRODUCTION: Pain is one of the main symptoms reported by patients who have had heart surgery. OBJECTIVES: To describe the pain and explain the possible association among demographics, psychological and biological variables of the patients subjected to heart surgery with pain intensity during the postoperative in the ICU. PATIENTS AND METHOD: A descriptive, longitudinal study conducted between February 2008 and January 2009 on patients subjected to heart surgery with admission to the ICU of the Hospital of Navarra was conducted. A preoperatory interview was made with registration of sociodemographic, biological and psychological variables. Pain intensity was monitored during the first 48 hours of ICU stay with the Verbal Numeric Scale (VNS) of pain. Accepted level of significance was p<0.05. RESULTS: A sample of 69 patients with mean age of 62, 26% women and 74% men was included. A superior statistical association was found between postoperative pain levels for age<65 years, bypass grafting with internal mammary artery and preoperatory anxiety variables. There was a significant increase in analgesic consumption for incomes>1400 ?/month, bypass grafting with internal mammary artery and preoperatory anxiety. CONCLUSIONS: Postoperative pain after heart surgery show significant individual variability. In our study, age, bypass grafting with internal mammary artery and preoperatory anxiety were shown as predictive variables of postoperative pain in patients undergoing heart surgery. PMID- 21640628 TI - Mannose-binding lectin serum levels in patients with leprosy are influenced by age and MBL2 genotypes. AB - BACKGROUND: Mannose-binding lectin (MBL) activates the complement system promoting opsonophagocytosis, which could represent an advantage for Mycobacterium leprae, an intracellular pathogen. Therefore, a single nucleotide polymorphism (SNP) in the MBL2 gene associated with low levels of MBL could confer protection against the development of leprosy disease. METHODS: In this study, we investigated SNPs of the MBL2 gene and MBL levels in 228 Brazilian leprosy patients and 232 controls. RESULTS: There were no differences in the frequencies of variant genotypes and haplotypes of MBL2 between patients and controls, or between the different clinical forms of leprosy. In the group of patients with a genotype for high expression of MBL2, those aged>40 years had decreased MBL levels compared to patients aged <= 40 years (p = 0.037). CONCLUSION: Our results demonstrate that age could influence the phenotype of MBL2, but no evidence was found for an association of MBL2 polymorphism with susceptibility to leprosy or its clinical forms. PMID- 21640629 TI - Differential responses of infectious diseases to economic growth in China. PMID- 21640630 TI - [Tuberculosis of the hand and wrist: different aspects of 30 cases]. AB - Tuberculosis of the hand and wrist is a rare entity. We report 30 cases of tuberculosis of the hand and wrist, including ten cases of wrist osteoarthritis, ten cases of tenosynovitis, four cases of metacarpophalangeal and interphalangeal osteoarthritis and six cases of metacarpal and phalangeal osteitis. The histological study after surgical biopsy revealed caseating giant cell granulomas with epitheloid cells confirming the diagnosis. Antibacillary chemotherapy promoted healing and good outcome in our patients. The aim of our work is to analyze the epidemiological, diagnostic, therapeutic and evolutionary aspects of this disease through a series of 30 cases. PMID- 21640631 TI - Restless legs syndrome in post-polio syndrome: a series of 10 patients with demographic, clinical and laboratorial findings. AB - BACKGROUND: Few studies have described the occurrence of restless legs syndrome in post-polio syndrome. METHODS: We studied 10 consecutive patients with post polio syndrome and symptoms of restless legs syndrome. We look at demographic, clinical and laboratorial data. RESULTS: A remarkable finding was the concomitant onset of symptoms of both diseases, suggesting a possible underlying mechanism. Severity of restless legs symptoms was moderate to very severe. CONCLUSION: Epidemiological studies with larger samples are needed to better establish the relationship and the incidence of restless legs syndrome in post-polio syndrome. PMID- 21640632 TI - Tree seasonality in a warming climate. AB - Climate warming has increased researchers' interest in plant phenology and its modelling. Although the main focus is on projections of accelerated springtime phenological events, also a further extension of the growing season by delayed growth cessation is often projected. However, ecophysiological studies indicate that, for boreal and temperate trees, such generalisations are precluded owing to differential climatic conditions and inter- and intraspecific genetic differences. The annual cycle of these trees is an integrated system, where one phase affects subsequent phases, resulting in delayed impacts, which are only partially addressed in current ecophysiological models. Here, we outline an updated integrated conceptual model of the annual cycle by identifying ecophysiological phenomena that are particularly significant under climate warming. PMID- 21640634 TI - IGF-1R expression predicts clinical outcome in patients with locally advanced oral squamous cell carcinoma. AB - To assess the expression of IGF-1R in oral cavity squamous cell carcinoma patients, to explore its relation with clinical and pathologic prognostic factors and its role in predicting clinical outcome. One hundred and thirty-one consecutive patients suffering from oral cavity squamous cell carcinoma were included in this study from July 1989 to April 2005. Follow-up was closed in May 2010. The mean follow-up for survivors was 110.26+/-47.42 months. Patients were staged following the TNM classification. Patients in tumour stages I and II were referred to surgery. Patients in stages III-IV were referred to postoperative radiotherapy. Radiation therapy was administered up to a mean dose of 62.13+/ 7.74 Gy in 1.8-2 Gy fractions. IGF-1R expression was studied by immunohistochemistry in paraffin-embedded tumour tissue. IGF-1R was expressed in 101 patients (77.1%). IGF-1R expression was related to tumour grade (P=0.012). Tumour stage was the most important prognostic factor for survival. Low (negative and fairly) IGF-1R tumour expression was correlated to better long-term Local Disease Free Survival (P=0.016), Disease-Free Survival (P=0.029), and Survival (P=0.009) in patients achieving tumour stages III-IV. Low IGF-1R expression was related to better long-term control in patients suffering locally advanced oral carcinoma. PMID- 21640633 TI - Translational imaging endpoints to predict treatment response to novel targeted anticancer agents. AB - Response Evaluation Criteria in Solid Tumors (RECIST) and World Health Organization (WHO) Criteria have been traditionally used for the evaluation of therapeutic response to chemotherapeutic treatment regimens. They determine anatomic criteria for patients response to anti-cancer therapy based on morphological measurements of each target lesion. While this assessment is justified for cytotoxic (chemotherapeutic) drugs, it is now recognized that morphological imaging protocols are poorly suited to the evaluation of the efficacy of novel signal transduction inhibitors (STIs) which exhibit cytostatic rather than cytotoxic properties. New imaging technologies are now designed to evaluate, in a functional manner, modifications in tumor metabolic activity, cellularity, and vascularization before a reduction in tumor volume can be detected. Introduction of physiological imaging end-points, derived from dynamic contrast-enhanced (DCE) imaging protocols--including magnetic resonance imaging (MRI), computed tomography (CT) and ultrasound (US)--allow for early assessment of disruption in tumor perfusion and permeability for targeted anti-angiogenic agents. Diffusion-weighted MRI (DWI) provides another physiological imaging end point since tumor necrosis and cellularity are seen early in response to anti angiogenic treatment. Changes in glucose and phospholipid turnover, based on metabolic MRI and positron emission tomography (PET), provide reliable markers for therapeutic response to novel receptor-targeting agents. Finally, novel molecular imaging techniques of protein and gene expression have been developed in animal models followed by a successful human application for gene therapy based protocols. PMID- 21640635 TI - Spectroscopic and molecular modeling evidence of clozapine binding to human serum albumin at subdomain IIA. AB - Various spectroscopy and molecular docking methods were used to examine the binding of Clozapine (CLZ) to human serum albumin (HSA) in this paper. By monitoring the intrinsic fluorescence of single Trp214 residue and performing Dansylamide (DNSA) displacement measurement, the specific binding of CLZ in the vicinity of Sudlow's Site I of HSA has been clarified. An apparent distance of 27.3 A between the Trp214 and CLZ was obtained via fluorescence resonance energy transfer (FRET) method. In addition, the changes in the secondary structure of HSA after its complexation with CLZ ligand were studied with CD spectroscopy, which indicate that CLZ does not has remarkable effect on the structure of the protein. Moreover, thermal denaturation experiment shows that the HSA-CLZ complexes are conformationally more stable. Finally, the binding details between CLZ and HSA were further confirmed by molecular docking studies, which revealed that CLZ was bound at subdomain IIA through multiple interactions, such as hydrophobic effect, van der Waals forces and hydrogen bonding. PMID- 21640636 TI - Synthesis, growth and characterization of cadmium manganese thiocyanate (CMTC) crystal. AB - Single crystals of cadmium manganese thiocyanate, CdMn(SCN)4 (CMTC) have been successfully synthesized and grown by slow evaporation solution growth technique using water as solvent at room temperature. The crystal was characterized by different techniques for finding its suitability for device fabrications. From the single crystal XRD the crystal system was identified as tetragonal. The functional groups were identified from FTIR analysis. The optical studies have been carried out and found that the tendency of transmission observed from the specimen with respect to the wavelength of light is practically more suitable for the present trends in communication engineering. From the thermal analysis the decomposing temperature of the grown crystal is more significant when compared with the studies performed earlier. PMID- 21640637 TI - Photophysical and theoretical insights on non-covalently linked fullerene-zinc phthalocyanine complexes. AB - The photo-physical aspects of non-covalently linked assemblies of a series of fullerenes, namely, C60, C70, tert-butyl-(1,2-methanofullerene)-61-carboxylate (1) and [6,6]-phenyl C70 butyric acid methyl ester (2) with a designed zinc phthalocyanine (ZnPc), viz., zinc-1,4,8,11,15,18,22,25-octabutoxy-29H,31H phthalocyanine (3) in toluene medium are studied employing absorption spectrophotometric, steady state and time resolved fluorescence spectroscopic measurements. Of central interest in these investigations is the preferential binding of various fullerenes with ZnPc in toluene. The ground state interaction between fullerenes and 3 is first evidenced from UV-Vis measurements. Steady state fluorescence experiment reveals efficient quenching of the excited singlet state of 3 in presence of both underivatized and derivatized fullerenes. K values for the complexes of C60, C70, 1 and 2 with 3 are determined to be 6500, 22,230, 47,800 and 54,770 dm3 mol(-1), respectively. The magnitude of K suggests that 3 preferentially binds C70 and derivatized C70 in comparison to C60 and 1. Time resolved emission measurements establish that C(70)-3 and 2-3 complexes are stabilized much more in comparison to C(60)-3 and 1-3 systems in terms of charge separation process. Semi empirical calculations employing third parametric method substantiate the strong binding of C70 and its derivative with 3 in terms of heat of formation values of the respective complexes, and at the same time, determine the orientation of bound guest (here fullerenes) with the molecular plane of 3. PMID- 21640638 TI - Density functional theory (DFT) investigation of molecular structure and frontier molecular orbitals (FMOs) of P-N,N-dimethylaminobenzylidenemalononitrile (DBM). AB - P-N,N-dimethylaminobenzylidenemalononitrile (DBM) dye belongs to a class of organic compounds known as molecular rotors. Its optimized geometry and frontier molecular orbitals (FMOs), before and after ultraviolet (UV) irradiation, were obtained by DFT/B3LYP level with complete relaxation in the potential energy surface using 6-311++G(d,p) basis set. It is found that the length of C-C bonds of the DBM molecule increases after the UV irradiation, which leads to an increase in its dipole moment making it as a promising material for solar cell applications. Also, its HOMO-LUMO gap decreased from 3.46 to 3.34 eV. From the cyclic voltammetry measurements the value of HOMO-LUMO gap is equal to 3.21 eV. This means that B3LYP/6-311++G(d,p) level of theory is the best one for calculations. PMID- 21640639 TI - Spectrophotometric studies on the thermodynamic properties of charge-transfer complexes between m-DNB (1,3-dinitrobenzene) with aliphatic amines in DMSO and determination of the vertical electron affinity of m-DNB. AB - 1,3-Dinitrobenzene formed colored 1:1 complexes with aliphatic amines (chromogenic agents) like isopropylamine,ethylenediamine, tetraethylenepentamine and bis(3-aminopropyl)amine in DMSO having absorption maxima at 563 nm, 584 nm, 580.5 nm and 555 nm respectively. The complexes were stable for more than 24 h. The accurate association constants KAD and other thermodynamic parameters were determined with D and A usually in stoichiometric ratios. But in case of m-DNB and bis(3-aminopropyl)amine, the association constants KAD and the thermodynamic parameters were also determined using Benesi-Hildebrand equation to show the variations of KAD under different conditions. DeltaG degrees values were found to be negative in all cases resulting from exothermic enthalpy changes and favourable entropy changes. The energies of transition for the CT complexes hnuCT found experimentally were considerably different from the energies of transition (from HOMO of donor to LUMO of acceptor) calculated using AM1 but the differences were considerably reduced using DFT calculations. The vertical electron affinity of m-DNB was calculated using the method suggested by Mulliken. However, no FTIR measurements of the complexes could be made due to experimental limitations. PMID- 21640640 TI - Spectral investigation of the intramolecular charge-transfer in some aminotriazole Schiff bases. AB - 3-Amino-1,2,4-triazole Schiff bases were reported to contain intramolecular charge-transfer. The enhancing and depressing effects were remarkable as the substituent was changed from electron-donating to electron-withdrawing groups. The path of the resonating delocalization was reversed in the case of the p-NO2 group. To validate these results we effectively used Weinhold et al's natural bond orbital analysis to assess the UV and FT-IR spectrophotometric monitoring of the change reflected in this phenomenon when the substituent in the benzene ring is altered. The NBO analysis was simulated by ab inito computations at the HF/6 31G(d) level of theory, in order to properly detect any possible presence of a hydrogen bond association. The changes occurring in electron occupancies of double-centered bonds, antibonding orbitals and in lone-pair orbitals appraised the results, as did the s and p character listings of the two-centered bonds and the simultaneous changes occurring in the geometric parameters of the molecules in question. Contrary to its normal preference, in these molecules the nitrogen used sp2 hybrid orbitals for its interaction, housing its electron lone-pair in the third p hybrid orbital. Furthermore, NBO analysis reflected the presence of a very soft intramolecular hydrogen association (C-H?pi), labelled by UV and FT-IR assignments, between the benzene and triazole rings in all Schiff bases but p N(Me)2. The n-pi* stabilization energy decreased in the order: p-OH>p-OCH3>p-Cl>p CH3>H>p-NO2>o-OH. The relation between the band position and Hammett substitution constant is interpreted in relation to the molecular structure. PMID- 21640641 TI - Phenoxide bridged tetranuclear Co(II), Ni(II), Cu(II) and Zn(II) complexes: electrochemical, magnetic and antimicrobial studies. AB - Phenoxide bridged later first row transition metal(II) complexes have been prepared by the interaction of later 3d transition metal(II) chlorides with tetranucleating compartmental Schiff base ligand system derived from 2,6-diformyl 4-methylphenol, p-phenylenediamine and 2-hydrazinobenzothiazole. Ligand and complexes were characterized by analytical, spectral (IR, UV-visible, ESR, FAB mass and fluorescence), magnetic and thermal studies. All complexes are found to have octahedral geometry. The mutual influence of metal centres in terms of cooperative effect on the electronic, magnetic, electrochemical and structural properties was investigated. The Schiff base and its complexes have been screened for their antibacterial (against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa) and antifungal activities (against Aspergillus niger, and Candida albicans). PMID- 21640642 TI - Speciation studies on the complex formation reactions of [Pd(N,N-diethyl ethylendiamine)(H2O)2]2+ with some bio-relevant ligands and displacement reaction by mercaptoethylamine. AB - Pd(deen)Cl2 and Pd(deen)(CBDCA) complexes, where deen=N,N-diethylethylenediamine and CBDCA=1,1-cyclobutanedicarboxylate, were synthesized and characterized by elemental analysis and spectroscopic techniques. The stoichiometry and stability of the complexes formed between various biologically relevant ligands (amino acids, peptides, DNA constituents and dicarboxylic acids) and [Pd(deen)(H2O)2]2+ were investigated at 25 degrees C and 0.1 M ionic strength. The speciation diagrams of the complexes formed in solutions are evaluated. The mode of coordination of glycylglycine is investigated by spectrophotometric measurements. The equilibrium constants for the displacement of coordinated ligands as inosine, glycine or methionine by mercaptoethylamine are calculated. The results are expected to contribute to the chemistry of antitumour agents. PMID- 21640643 TI - Health information seeking, diet and physical activity: an empirical assessment by medium and critical demographics. AB - PURPOSE: Research over the past decade has conveyed a dramatic rise in health information seeking via the Internet and articulated various profiles and outcomes of health information seeking. In building upon this research, the current study is innovative in considering predictors of health information seeking by medium, as well as outcomes of health information seeking by medium and by critical demographics. METHODS: OLS regression and logistic regression are conducted on data from a telephone survey of American adults in 2007 (N=700). RESULTS: Profiles of health information seekers vary dramatically by medium (Internet versus newspapers versus television). In terms of outcomes, newspaper health information seeking is associated with fruit and vegetable consumption, while television health information seeking is associated with sweetened soft drink consumption. There are four significant interaction terms between these two health information seeking variables and unhealthy snack consumption. Internet health information seeking has no significant effects. CONCLUSIONS: In comparison to the Internet, newspaper and television media have more favorable associations with recommended levels of lifestyle behaviors that may be critical in efforts to decrease obesity in the United States. PMID- 21640644 TI - A diagnostic and public health quandary: acute viral hepatitis in a hospital cafeteria worker. PMID- 21640645 TI - Unique post-exercise electrophysiological test results in a new Andersen-Tawil syndrome mutation. PMID- 21640646 TI - Ocular vestibular evoked myogenic potentials produced by impulsive lateral acceleration in unilateral vestibular dysfunction. AB - OBJECTIVE: To deduce the connectivity underlying ocular vestibular evoked myogenic potentials (OVEMPs) recorded from two sites and produced by lateral transmastoid stimulation in patients with unilateral vestibular dysfunction. METHODS: OVEMPs were recorded using lateral impulsive stimuli delivered by a hand held minishaker placed at the mastoid. Twelve patients were tested using the typical OVEMP recording montage placed inferior to the eyes. In a subset of 6 patients, recordings were also made using a lateral electrode montage. The majority of patients were tested following surgery for inner ear disease. Patient responses were compared to those in normal subjects under similar recording conditions. RESULTS: For the inferior montage, regardless of which mastoid was stimulated, deficits were observed only from the eye opposite the affected ear. In contrast, OVEMPs recorded using the lateral electrode montage showed changes on both sides. CONCLUSIONS: OVEMPs produced using lateral transmastoid stimulation and recorded from beneath the eyes are generated by a crossed vestibulo-ocular pathway while the projections underlying the lateral responses are likely to be bilateral. SIGNIFICANCE: The vestibular-ocular connectivity underlying the OVEMPs recorded from inferior and lateral recording sites differs. For clinical use, the inferior recording site is the simplest to interpret. PMID- 21640647 TI - Application of Fourier transform near-infrared spectroscopy to optimization of green tea steaming process conditions. AB - In this study, we constructed prediction models by metabolic fingerprinting of fresh green tea leaves using Fourier transform near-infrared (FT-NIR) spectroscopy and partial least squares (PLS) regression analysis to objectively optimize of the steaming process conditions in green tea manufacture. The steaming process is the most important step for manufacturing high quality green tea products. However, the parameter setting of the steamer is currently determined subjectively by the manufacturer. Therefore, a simple and robust system that can be used to objectively set the steaming process parameters is necessary. We focused on FT-NIR spectroscopy because of its simple operation, quick measurement, and low running costs. After removal of noise in the spectral data by principal component analysis (PCA), PLS regression analysis was performed using spectral information as independent variables, and the steaming parameters set by experienced manufacturers as dependent variables. The prediction models were successfully constructed with satisfactory accuracy. Moreover, the results of the demonstrated experiment suggested that the green tea steaming process parameters could be predicted on a larger manufacturing scale. This technique will contribute to improvement of the quality and productivity of green tea because it can objectively optimize the complicated green tea steaming process and will be suitable for practical use in green tea manufacture. PMID- 21640648 TI - Growth factor/heparin-immobilized collagen gel system enhances viability of transplanted hepatocytes and induces angiogenesis. AB - Hepatocyte transplantation is being explored as a treatment strategy for end stage liver disease; however, the main limitation is the insufficient vascularization of transplanted hepatocytes. To overcome this problem, a suitable 3D microenvironment and the types of transplanted cells must be considered for hepatocyte transplantation. In this study, a growth factor (GF)/heparin immobilized collagen gel-filled polyurethane foam (PUF) scaffold was developed for angiogenesis induction and hepatocyte transplantation. First, a vascular endothelial growth factor (VEGF)/heparin-immobilized, collagen-gel-filled PUF scaffold was developed to establish a prevascularized cavity in the subcutaneous space in rats. Second, accompanied by 70% partial hepatectomy (PH), hepatocytes were embedded inside heparin-immobilized, collagen-gel-filled PUF scaffolds, and were transplanted into the VEGF-induced prevascularized cavity. The benefits of using this system were confirmed by using three types of hepatocytes, namely single hepatocyte, hepatocyte spheroids, and fetal hepatocytes. The normalized hemoglobin content and live nucleus numbers were determined separately to evaluate the angiogenesis and viability of transplanted hepatocytes. In summary, after PH pretreatment, transplantation of fetal hepatocyte-embedded, heparin immobilized, collagen-gel-filled PUF scaffold into a VEGF-induced prevascularized cavity appears to be a promising strategy for future liver tissue engineering. PMID- 21640649 TI - Isokinetic dynamometry applied to shoulder rotators - velocity limitations in eccentric evaluations. AB - The objectives of this study were to evaluate if collegiate overhead athletes, with and without shoulder pain, and non-athletes could reach a preset velocity in internal and external shoulder rotation isokinetic evaluations; and to evaluate the correlation between torque and velocity. Controlled laboratory study, cross sectional. Evaluations were performed using the isokinetic dynamometer Biodex System 3. Participants were assessed seated, with the arm at 90 degrees of shoulder abduction and 90 degrees of elbow flexion, from neutral rotation to 90 degrees of external rotation. Five maximal contractions of isokinetic concentric and reactive eccentric internal and external rotation were performed at the velocities 90 degrees /s, 180 degrees /s and 240 degrees /s. Data were processed with using MatLab. Most participants did not reach the isokinetic phase during eccentric tests at 180 degrees /s and 240 degrees /s, particularly in the external rotators evaluation. High correlations between torque and velocity of eccentric tests were found. The groups presented no differences in maximal velocity attained in trials which preset velocity was not reached. These results call into question the use of reactive eccentric tests at velocities higher than 180 degrees /s for the isokinetic evaluation of shoulder external rotators in collegiate overhead athletes and non-athletes in this specific position. In such cases, careful evaluation of the velocity is recommended to determine if the isokinetic phase was reached. PMID- 21640650 TI - Recurrent cardiac constriction after complete pericardiectomy. AB - We present a patient with recurrent constrictive physiology resulting from exuberant post-operative fibrosis after complete pericardiectomy. The patient underwent a repeat stripping procedure. At surgery, there was an extensive fibrotic and calcified rind around the heart. The recurrence of constriction physiology after complete pericardiectomy in non-tuberculous pericarditis is a rarely reported in literature. The management supports repeat surgery and the potential value of steroid administration. PMID- 21640651 TI - Clazosentan, an endothelin receptor antagonist, in patients with aneurysmal subarachnoid haemorrhage undergoing surgical clipping: a randomised, double blind, placebo-controlled phase 3 trial (CONSCIOUS-2). AB - BACKGROUND: Clazosentan, an endothelin receptor antagonist, significantly and dose-dependently reduced angiographic vasospasm after aneurysmal subarachnoid haemorrhage (aSAH). We investigated whether clazosentan reduced vasospasm-related morbidity and all-cause mortality. METHODS: In this randomised, double-blind, placebo-controlled, phase 3 study, we randomly assigned patients with aSAH secured by surgical clipping to clazosentan (5 mg/h, n=768) or placebo (n=389) for up to 14 days (27 countries, 102 sites, inpatient and outpatient settings) using an interactive web response system. The primary composite endpoint (week 6) included all-cause mortality, vasospasm-related new cerebral infarcts, delayed ischaemic neurological deficit due to vasospasm, and rescue therapy for vasospasm. The main secondary endpoint was dichotomised extended Glasgow outcome scale (GOSE; week 12). This trial is registered with ClinicalTrials.gov, number NCT00558311. FINDINGS: In the all-treated dataset, the primary endpoint was met in 161 (21%) of 764 clazosentan-treated patients and 97 (25%) of 383 placebo treated patients (relative risk reduction 17%, 95% CI -4 to 33; p=0.10). Poor functional outcome (GOSE score <=4) occurred in 224 (29%) clazosentan-treated patients and 95 (25%) placebo-treated patients (-18%, -45 to 4; p=0.10). Lung complications, anaemia, and hypotension were more common with clazosentan. Mortality (week 12) was 6% in both groups. INTERPRETATION: Clazosentan at 5 mg/h had no significant effect on mortality and vasospasm-related morbidity or functional outcome. Further investigation of patients undergoing endovascular coiling of ruptured aneurysms is needed to fully understand the potential usefulness of clazosentan in patients with aSAH. FUNDING: Actelion Pharmaceuticals. PMID- 21640652 TI - Secondary brain injury after aneurysmal subarachnoid haemorrhage: more than vasospasm. PMID- 21640653 TI - The impact of discharge plan upon re-admission, satisfaction with nursing care and the ability to self-care for coronary artery bypass graft surgery patients. AB - BACKGROUND AND AIM: Grafting coronary arteries and post operative recovery has many challenges, which can be ameliorated through continues care and an appropriate discharge plan. Therefore, the current study was undertaken aiming to evaluate the impact of discharge plan on satisfaction with nursing care, ability to self-care, and incidence of re-admission. METHOD: This is a quasi experimental study involving patients who were due to undergo coronary artery bypass graft in Chamran Hospital in 2010. In the intervention group, the discharge plan was initiated at the time of admission and continued for 2 weeks after discharge by home visit and telephone follow ups. Satisfaction with nursing care was assessed 2 days after discharge, whilst patients' ability for self-care was measured 6 weeks and 3 months post discharge and the incidence of re-admission was determined at the 3 months point. FINDINGS: Satisfaction levels with nursing care and the ability to take self-care were higher in intervention group comparing with control group (p < 0.001). There was a significant difference for self-care ability between pre test and post test in both groups but the improvement was more pronounced for the intervention group (p = 0.04). There was no significant difference between the two groups in terms of re-admission incidence after 3 months (p = 0.15). DISCUSSION AND CONCLUSION: The results indicate that the discharge plan, as a method of continual care plan, can lead to higher satisfaction levels and enhanced self-care abilities of patients. Such discharge plan can therefore be utilised as an effective method of continuous care for patients who are going to undergo coronary artery bypass graft. PMID- 21640654 TI - Relationships among vesicoureteric reflux, urinary tract infection and renal injury in children with non-neurogenic lower urinary tract dysfunction. AB - OBJECTIVE: To determine the relationship between vesicoureteric reflux (VUR), urinary tract infection (UTI), renal damage and the pattern of non-neurogenic lower urinary tract dysfunction (LUTD), and to reveal the possible risk factors for renal damage in children with LUTD. METHODS: For the years 2004-2010, demographic, clinical, laboratory and urodynamic study reports of children with LUTD were retrospectively reviewed. RESULTS: Of 96 patients, there were diagnosed 70 with overactive bladder (OAB), 8 pure dysfunctional voiding (DV) and 18 OAB plus DV. The rate of VUR, UTI and renal damage in patients with OAB plus DV and pure DV was higher than in patients with OAB alone. VUR was significantly higher among the patients who had UTI. Renal scarring was detected in 25 patients, of whom 78% had OAB plus DV and 75% DV. The presence of VUR was associated with a significant increase in the rate of renal damage, and dilating reflux caused significantly greater damage compared to non-dilating reflux. CONCLUSION: OAB plus DV and DV are major risk factors for VUR, UTI and renal damage. The presence of VUR in children with LUTD plays an important role with regard to UTI and renal damage, with dilating VUR a major risk factor associated with renal damage. PMID- 21640655 TI - Benchmarking and patient safety in hypertensive disorders of pregnancy. AB - Hypertensive disorders of pregnancy are a major cause of morbidity and mortality worldwide. Reliable, published individual patient data from units and countries are lacking. Without these data, clinicians are unable to benchmark their incidence, treatments and outcomes, and patient safety is unable to be routinely assessed. Available data suggest that a notable proportion of the adverse events that occur with hypertensive disease of pregnancy may be preventable. Theory and practice indicate several methods that can offer the possibility of averting these preventable adverse events. These methods include benchmarking outcomes, standardisation of care processes, simulation, and enhancement of patient knowledge. However, data on optimal methods to enhance patient safety and quality of care of pregnant women with hypertensive disease remain limited, and further research is required. PMID- 21640656 TI - Which score most likely represents pain on the observational PAINAD pain scale for patients with dementia? AB - OBJECTIVES: We sought to determine a cutoff score for the observational Pain Assessment in Advanced Dementia (PAINAD), to adequately assess pain in clinical nursing home practice and research. DESIGN AND SETTING: We used data from multiple sources. We performed a literature review on PAINAD, performed secondary data analysis of a study examining psychometric properties of PAINAD in nursing home patients with dementia, and performed another study in nursing home patients with dementia specifically aimed at determining a cutoff score for PAINAD. PARTICIPANTS: Patients with dementia in long term care facilities. MEASUREMENTS: We related PAINAD scores (range 0 to 10) to (1) self-reported and proxy-reported pain by global clinical judgment and (2) scores on another pain assessment instrument (DOLOPLUS-2), and (3) we compared scores between painful and supposedly less painful conditions. RESULTS: Findings from this study showed that a cutoff value of 2 should serve as a trigger for a trial with pain treatment. Although the majority of patients scoring 1 or 0 were not in pain, pain could be ruled out. CONCLUSION: Based on the findings of multiple available data sources, we recommend that a PAINAD score of 2 or more can be used as an indicator of probable pain. A score of 1 is a sign to be attentive to possible pain. Future work may focus on cutoff scores for the presence of pain and severe pain in other frequently used pain tools, and on further development of methodology to assess cutoff scores. PMID- 21640658 TI - Evaluation and management of combat-related spinal injuries: a review based on recent experiences. AB - BACKGROUND CONTEXT: The current approach to the evaluation and treatment of military casualties in the Global War on Terror is informed by medical experience from prior conflicts and combat encounters from the last 10 years. In an effort to standardize the care provided to military casualties in the ongoing conflicts, the Department of Defense (DoD) has published Clinical Practice Guidelines (CPGs) that deal specifically with the combat casualty sustaining a spinal injury. However, the combat experience with spine injuries in the present conflicts remains incompletely described. PURPOSE: To describe the CPGs for the care of the combat casualty with suspected spine injuries and discuss them in light of the published military experience with combat-related spinal trauma. STUDY DESIGN: Literature review. METHODS: A literature review was conducted regarding published works that discussed the incidence, epidemiology, and management of combat related spinal trauma. The CPGs, established by the DoD, are discussed in light of actual military experiences with spine trauma, the present situation in the forward surgical teams and combat support hospitals treating casualties in theater, and recent publications in the field of spine surgery. RESULTS: In the conventional wars fought by the United States between 1950 and 1991 (Korea, Vietnam, Gulf War I), the incidence of spine injuries remained close to 1% of all combat casualties. However, in the Global War on Terror, the enemy has relied on implements of asymmetric warfare, including sniper attacks, ambush, roadside bombs, and improvised explosive devices. The increase in explosive mechanisms of injury has elevated the number of soldiers exposed to blunt force trauma and, consequently, recent publications reported the highest incidence of combat related spinal injuries in American military history. Wounded soldiers are expeditiously evacuated through the echelons of care but typically do not receive surgical management in theater. The current CPGs for the care of soldiers with combat-related spinal injuries should be re-examined in light of data regarding the increasing number of spine injuries, new injury patterns, such as lumbosacral dissociation and low lumbar burst fractures, and recent reports within the field of spine surgery as a whole. CONCLUSIONS: American and coalition forces are sustaining the highest spine combat casualty rates in recorded history and previously unseen injuries are being encountered with increased frequency. While the CPGs provide useful direction in terms of the evaluation and management of combat casualties with spine injuries, such recommendations may warrant periodic re-evaluation in light of recent combat experiences and evolving scientific evidence within the spine literature. PMID- 21640659 TI - Natural history of spinal deformity in a patient with Ehlers-Danlos syndrome: case report with 20-year follow-up. AB - BACKGROUND CONTEXT: Scoliosis can present in patients with Ehlers-Danlos syndrome (EDS) and can be surgically treated despite possible complications. The long-term natural history of the progression of spinal deformity, particularly scoliosis, and associated complications in EDS patients remains relatively unknown. PURPOSE: To assess the progression of spinal deformity and associated complications in a patient with EDS, who was not surgically treated for her spine condition and was followed up for 20 years. STUDY DESIGN: A case report. METHODS: A 9-year-old female with EDS presented to the orthopedic clinic with scoliosis and mild thoracic hyperkyphosis. The patient was followed up for a 20-year period. Progression of her spinal deformity and associated complications were noted. RESULTS: The patient's curve progressed rapidly in the initial follow-up period an 83 degrees worsening from the age of 9 to 10 years. Because she refused surgical intervention, her spinal deformity continued to progress. As such, by the age of 29 years, she presented with 115 degrees from T9 to L4 with severe hyperkyphosis. The patient eventually developed respiratory compromise and diminished functional capacity. CONCLUSIONS: In patients with EDS, scoliosis may progress rapidly and unrelentingly, and if not treated early may lead to respiratory compromise. Close follow-up of such patients is imperative, and appropriate management should be sought to prevent potentially fatal morbidities. PMID- 21640657 TI - Sarcopenia with limited mobility: an international consensus. AB - A consensus conference convened by the Society of Sarcopenia, Cachexia and Wasting Disorders has concluded that "Sarcopenia, ie, reduced muscle mass, with limited mobility" should be considered an important clinical entity and that most older persons should be screened for this condition. "Sarcopenia with limited mobility" is defined as a person with muscle loss whose walking speed is equal to or less than 1 m/s or who walks less than 400 m during a 6-minute walk, and who has a lean appendicular mass corrected for height squared of 2 standard deviations or more below the mean of healthy persons between 20 and 30 years of age of the same ethnic group. The limitation in mobility should not clearly be a result of otherwise defined specific diseases of muscle, peripheral vascular disease with intermittent claudication, central and peripheral nervous system disorders, or cachexia. Clinically significant interventions are defined as an increase in the 6-minute walk of at least 50 meters or an increase of walking speed of at least 0.1 m/s. PMID- 21640660 TI - A comparison of biomechanical stability and pullout strength of two C1-C2 fixation constructs. AB - BACKGROUND CONTEXT: Several fusion techniques are used to treat atlantoaxial instability. Recent literature suggests that intralaminar screw (LS) fixation and pedicle screw (PS) fixation offer similar stability and comparable pullout strength. No studies have compared these characteristics after cyclic loading. PURPOSE: To compare the stability and pullout strength of intra-LSs and PSs in a C1-C2 instability model after 1,000 cycles of axial loading. STUDY DESIGN: In vitro biomechanical study. OUTCOME MEASURES: Stability in axial rotation and screw pullout strength after cyclic loading. METHODS: Six fresh-frozen human cadaveric cervical spines (C1-C2) were used in this study. C1-C2 instability was mimicked via odontoidotomy at its base and posterior soft-tissue release, including the supraspinous ligaments and facet joint capsules. Specimens were tested to 1,000 cycles after stabilization with two fixation constructs: C1 lateral mass (LM) screws and C2 intra-LSs (C1LM-C2LS) and C1 LM screws and C2 PSs (C1LM-C2PS). Angular motion was recorded for right and left axial rotation using an Optotrak 3020 system (Northern Digital, Waterloo, Ontario, Canada). Tensile loading to failure was then performed collinear to the longitudinal axis of the screw, and the data were recorded as peak pullout strength in newtons. RESULTS: There was no statistically significant difference in stability (measured in degrees of rotation) between the intra-LS and PS constructs at 250, 500, 750, and 1,000 cycles of axial rotation. Furthermore, there was no significant difference in stability at 250 cycles versus 1,000 cycles for the LS (1.30 vs. 1.49, p = .80) or PS (0.84 vs. 0.85, p = .96). Pedicle screws had higher pullout strength when compared with the intra-LSs (757.5 +/- 239 vs. 583.4 +/- 472 N); however, high standard deviation precluded statistical significance (p = .44). CONCLUSIONS: Our data suggest that a C1LM and C2LS construct has similar biomechanical stability when compared with a C1LM and C2PS construct after 1,000 cycles of axial rotation. Furthermore, PSs had higher pullout strength when compared with LSs; however, this result was not statistically significant. PMID- 21640661 TI - In vivo diode dosimetry vs. computerized tomography and digitally reconstructed radiographs for critical organ dose calculation in high-dose-rate brachytherapy of cervical cancer. AB - PURPOSE: To investigate the correlation between the dose predicted by the treatment planning system using digitally reconstructed radiographs or three dimensional (3D)-reconstructed CT images and the dose measured by semiconductor detectors, under clinical conditions of high-dose-rate brachytherapy of the cervix uteri. PATIENTS AND METHODS: Thirty-two intracavitary brachytherapy applications were performed for 12 patients with cancer of the cervix uteri. The prescribed dose to Point A was 7 Gy. Dose was calculated for both International Commissioning on Radiation Units and Measurements (ICRU) bladder and rectal points based on digitally reconstructed radiographs and for 3D CT images-based volumetric calculation of the bladder and rectum. In vivo diode dosimetry was performed for the bladder and rectum. RESULTS: The ICRU reference point and the volumes of 1, 2, and 5cm(3) received 3.6+/-0.9, 5.6+/-2.0, 5.1+/-1.7, 4.3+/-1.4 and 5.0+/-1.2, 5.3+/-1.3, 4.9+/-1.1, and 4.2+/-0.9 Gy for the bladder and rectum, respectively. The ratio of the 1cm(3) and the ICRU reference point dose to the diode dose was 1.8+/-0.7 and 1.2+/-0.5 for the bladder and 1.9+/-0.6 and 1.7+/ 0.5 for the rectum, respectively. CONCLUSIONS: 3D image-based dose calculation is the most accurate and reliable method to evaluate the dose given to critical organs. In vivo diode dosimetry is an important method of quality assurance, but clinical decisions should be made based on 3D-reconstructed CT image calculations. PMID- 21640662 TI - Whitmore, Henschke, and Hilaris: The reorientation of prostate brachytherapy (1970-1987). AB - PURPOSE: Urologists had performed prostate brachytherapy for decades before New York's Memorial Hospital retropubic program. This paper explores the contribution of Willet Whitmore, Ulrich Henschke, Basil Hilaris, and Memorial's physicists to the evolution of the procedure. METHODS AND MATERIALS: Literature review and interviews with program participants. RESULTS: More than 1000 retropubic implants were performed at Memorial between 1970 and 1987. Unlike previous efforts, Memorial's program benefited from the participation of three disciplines in its conception and execution. CONCLUSIONS: Memorial's retropubic program was a collaboration of urologists, radiation therapists, and physicists. Their approach focused greater attention on dosimetry and radiation safety, and served as a template for subsequent prostate brachytherapy programs. PMID- 21640664 TI - Pain and symptom assessment during multiple fractions of gynecologic high-dose rate brachytherapy. AB - PURPOSE: A prospective assessment of tolerability of gynecologic brachytherapy was completed to determine adequacy of analgesia and symptom control for patients undergoing CT-guided brachytherapy, with multiple fractions delivered during a single applicator insertion. METHODS AND MATERIALS: Seventeen patients receiving high-dose-rate brachytherapy for gynecologic cancer (other than vaginal vault) completed ratings of pain intensity, anxiety, and nausea at five key time points before, during, and after brachytherapy. Symptoms were assessed with patient reported scores using an 11-point numeric rating scale. The patient population included cervical (n=12), endometrial (n=3), and vulvar-vaginal (n=2) malignancies. Patients underwent general anesthesia for applicator placement. Analgesia consisted of subcutaneous route opioid, and oral opioid and/or nonopioid as needed for the duration of the treatment planning and delivery. RESULTS: The mean scores for pain were highest after patients were transferred to the CT scanner, 3.3+/-2.6, compared with baseline scores of 0.9+/-1.7. Pain scores were 2.3+/-2.3 during the remainder of the procedure, and 2.7+/-2.1 after the removal of the applicator. The highest mean anxiety scores occurred before the brachytherapy procedure, 4.3+/-3.4, with resolution of anxiety during the procedure to 1.3+/-1.6. The mode of nausea scoring during the procedure was 0. CONCLUSION: For most of the patients, the delivery of multiple fractions of image guided high-dose-rate brachytherapy is well tolerated with maximum scores of mild moderate pain and distress, and no significant nausea. This can be accomplished with applicator placement under general anesthesia and standard medical management. PMID- 21640663 TI - Cesium-131 permanent seed brachytherapy: dosimetric evaluation and radiation exposure to surgeons, radiation oncologists, and staff. AB - PURPOSE: Cesium-131 ((131)Cs) radioactive seed is Food and Drug Administration approved for permanent seed implant for all cancers, including lung and head and neck (HN) cancers. We describe the first clinical report of (131)Cs dosimetry and exposure rates to treating physicians and staff. METHODS AND MATERIALS: Twenty eight patients received (131)Cs implant for early stage lung and recurrent HN cancers. A nomogram was developed to calculate the number of seeds needed to cover the wedge line with the prescription dose (80 Gy). Final dosimetry was obtained after CT planning a few days following the surgical procedure. Radiation exposure to the treating physicians and staff was measured at the completion of the procedure. RESULTS: A nomogram was developed using the variseed software with source data from American Association of Physicists in Medicine TG-43 report. The total volume covered by the prescription isodose line of (131)Cs was measured and compared with (125)I. The prescription volume was smaller for (131)Cs. In addition, the exposure rate with (131)Cs was found to be acceptable. CONCLUSION: Our preliminary experience with (131)Cs lung and HN brachytherapy has been very encouraging with excellent dosimetric coverage and acceptable exposure to the treating physicians and staff. PMID- 21640665 TI - Stoma size critical to 12-month outcomes in endoscopic suturing for gastric bypass repair. AB - BACKGROUND: Roux-en-Y gastric bypass (RYGB) is the most frequently performed bariatric procedure. However, weight regain after RYGB is common and often associated with pouch and stoma dilation. Historically, revision surgery has a greater risk of morbidity and mortality than the primary procedure. Endoscopic repair appears to be a safer option; however, current knowledge is limited regarding the longer term outcomes. Our objective was to prospectively collect the 12-month post-RYGB outcomes data after repair of dilated gastric tissue with an incisionless tissue approximation system in an open-label, single-group study at 9 U.S. sites. METHODS: Adults >= 2 years after RYGB, with weight regain and pouch and/or stoma dilation underwent tissue plication with an endolumenal anchoring system to tighten dilated gastric tissue. The outcomes were captured, with statistical modeling used to identify the predictors of success. RESULTS: Of the 116 subjects, 112 (97%) had anchors successfully placed (mean 5.9 anchors/subject). The mean stoma diameter and pouch length after the procedure was 11.5 mm (50% reduction) and 3.3 cm (44% reduction), respectively. At 12 months after repair (n = 73), the mean weight loss and percentage of excess weight loss was 5.9 +/- 1.1 kg and 14.5% +/- 3.1%, respectively. Anchor presence was confirmed endoscopically in 61 (92%) of 66 patients at 1 year. Those with a dilated stoma (>12 mm) who had a postrepair diameter of <10 mm (n = 22, 30% of 66) had more than double the excess weight loss compared with the rest of the cohort (24% versus 10%, P = .03). No serious adverse events occurred. CONCLUSION: The 12-month outcomes have demonstrated the safety and durability of this method of gastric bypass repair. Aggressive reduction of stoma dilation was associated with superior weight loss. PMID- 21640666 TI - Who is the man in the mirror? Depersonalization disorder after obesity surgery. PMID- 21640667 TI - Predictors of psychological symptoms in morbidly obese patients after gastric bypass surgery. AB - BACKGROUND: Morbid obesity is associated with debilitating psychosocial consequences, such as depression, anxiety, and low self-esteem. One of the main goals of bariatric surgery should not only be reducing weight and counteracting co-morbid conditions, but also improving postoperative psychosocial functioning. The objective of our study was to determine the preoperative variables that could predict the psychological symptoms 6 and 12 months after surgery to improve the clinical outcome of morbidly obese patients undergoing bariatric surgery. The setting was a university hospital in Spain. METHODS: The study group consisted of 60 morbidly obese patients (46 women and 14 men) who had undergone gastric bypass surgery for weight reduction and had >= 1 year of follow-up. The patients were evaluated using different questionnaires (Symptom Checklist-90-Revised, Body Shape Questionnaire, Rosenberg Self-Esteem Questionnaire, COPE, Medical Outcomes Study Social Support Survey) at 3 points: before surgery, at 6 months postoperatively, and at 1 year postoperatively. To determine the influence of preoperative variables on long-term psychological status, a step-by-step multivariate linear regression analysis was performed, taking P < .05 as statistically significant. RESULTS: The variables of self-esteem, instrumental support, and coping focused in the emotions were the main preoperative predictors for psychological symptoms 6 months after surgery. Preoperative drug abuse as a strategy of problem solving and mainly body image significantly predicted most of the psychological disorders 1 year after surgery. CONCLUSION: The study of different psychosocial variables in patients before bariatric surgery is an important aid to predict postoperative psychological functioning. Self-esteem and body image were the most influential factors in the postoperative psychological outcome of morbidly obese patients in our study. PMID- 21640668 TI - Comment on: Novel bariatric technology: laparoscopic adjustable gastric banded plication: technique and preliminary results. PMID- 21640669 TI - Minilaparoscopic versus conventional laparoscopic hysterectomy: results of a randomized trial. AB - STUDY OBJECTIVE: To compare operative outcomes and postoperative pain of laparoscopic hysterectomy (LH) versus minilaparoscopic hysterectomy (MLH). DESIGN: Randomized controlled trial (Canadian Task Force Classification I). SETTING: Tertiary care center. PATIENTS: Seventy-six women scheduled to undergo a hysterectomy for a supposed benign gynecologic condition. INTERVENTIONS: Participants were randomly assigned to LH (n = 38) or MLH (n = 38). MLH was performed with use of 3-mm ports. Both patients and assessors of the postoperative outcomes were blinded to the size of port used, and patients' wounds were concealed by standard-size nontransparent dressings. MEASUREMENTS: Primary outcome was postoperative pain (both rest and incident on coughing and abdominal pain, as well as shoulder pain) by use of a 100-mm visual analogue scale. MAIN RESULTS: The two groups were similar in terms of operative outcomes. No intraoperative conversion from MLH to both LH and open surgery occurred. No significant difference in pain scores at 1, 3, 8, and 24 hours after surgery between groups was found. Rescue analgesic requirement was similar in the MLH and LH groups (21.1% vs 13.2%, p =.54). CONCLUSIONS: Ports can safely be reduced in size without a negative impact on the surgeon's ability to perform LH. MLH appears to have no advantage over LH in terms of postoperative pain. PMID- 21640670 TI - Poly-glutamic acid modified carbon nanotube-doped carbon paste electrode for sensitive detection of L-tryptophan. AB - A novel poly-glutamic acid (PGA) film modified carbon paste electrode (CPE) incorporating carbon nanotubes (CNTs) was first prepared for the determination of l-tryptophan (l-Trp). Scanning electron microscopy and Fourier transform infrared spectroscopy were applied for characterization of the surface morphology of the modified electrodes and cyclic voltammetry was used to investigate the electrochemical properties of the proposed electrode towards the oxidation of l Trp. Optimization of the experimental parameters was performed with regard to pH, ratio of CNTs, concentration of glutamic acid, electro-polymerization cycles, accumulation time and concentration of sodium dodecylbenzene sulfonate. The linearity between the oxidation peak current and the l-Trp concentration was obtained in the range of 5.0*10(-8) to 1.0*10(-4)M with a detection limit of 1.0*10(-8)M (S/N=3) and the sensitivity was calculated to be 1143.79MUA?mM( 1)?cm(-2). In addition, the PGA modified CPE incorporating CNTs displayed high selectivity, good stability and reproducibility, making it suitable for the routine analysis of l-Trp in clinical use. PMID- 21640671 TI - Tall claims: Mortality selection and the height of children in India. AB - Data from three rounds of nationally representative health surveys in India (1992/93, 1998/99 and 2005/06) are used to assess the impact of selective mortality on children's anthropometrics. The nutritional status of the child population was simulated under the counterfactual scenario that all children who died in the first three years of life were alive at the time of measurement. The simulations demonstrate that the difference in anthropometrics due to selective mortality would be large only if there were very large differences in anthropometrics between the children who died and those who survived. Differences of this size are not substantiated by the research on the degree of association between mortality and malnutrition. The study shows that although mortality risk is higher among malnourished children, selective mortality has only a minor impact on the measured nutritional status of children stratified by gender. PMID- 21640672 TI - The meanings of musical meanings: Comment on "Towards a neural basis of processing musical semantics" by Stefan Koelsch. PMID- 21640673 TI - Sedation and monitoring for gastrointestinal endoscopy: A nationwide web survey in Italy. AB - BACKGROUND: Best strategy of sedation/analgesia in gastrointestinal (GI) endoscopy is still debated. AIMS OF THE STUDY: To evaluate sedation and monitoring practice among Italian gastroenterologists and to assess their opinion about non-anaesthesiologist propofol administration. METHODS: A 19-item survey was mailed to all 1192 members of the Italian Society of Digestive Endoscopy (SIED). For each respondent were recorded demographic data, medical specialty, years of practise and practise setting. RESULTS: A total of 494 SIED members returned questionnaires, representing a response rate of 41.4%. The most employed sedation pattern was benzodiazepines for oesophagogastroduodenoscopies (EGDS) in 50.8% of procedures, benzodiazepines plus opioids for colonoscopy and enteroscopy in 39.5% and 35.3% of procedures, respectively, propofol for endoscopic retrograde colangiopancreatography (ERCP) and endoscopic ultrasound (EUS) in 42.3% and 35.6% of procedures, respectively. With regard to propofol use, 66% respondents stated that propofol was exclusively administered by anaesthesiologists. However, 76.9% respondents would consider non anaesthesiologist propofol administration after appropriate training. Pulse oximetry is the most employed system for procedural monitoring. Supplemental O(2) is routinely administered by 39.3% respondents. CONCLUSIONS: Use of sedation has become a standard practise during GI endoscopy in Italy. Pattern varies for each type of procedure. Pulse oximetry is the most employed system of monitoring. Administration of propofol is still directed by anaesthesiologists. PMID- 21640674 TI - Developmental aspects of respiratory chain from fetus to infancy. AB - Reviewing the recent literature on the role of mitochondria during fetal development paradoxically reveals two features: the importance of mitochondria in these early developmental phases, and the scarcity of information available for humans. Indeed, most of the available information on the role of mitochondria during development comes from studies of animal models that do not necessarily strictly apply to humans. In this paper, we attempted to collect information existing on humans, together with data from animal studies essentially presented as corroboration. This makes clear that a complex interacting network of energetic, genetic and epigenetic factors governs the impact of mitochondrial function on early development in humans. This complexity presumably also accounts for our poor understanding of the consequences of impaired mitochondrial function on prenatal development, or conversely, of the impact of development on the expression of such deficiencies. PMID- 21640675 TI - Flexi-Seal tube use for enteric fistula control in abdominal wall reconstruction- a response. PMID- 21640676 TI - Anomalous left-to-right shunting communication between the ascending aorta and right pulmonary artery in a dog. AB - Anomalies of conotruncal septation are rare in dogs and uncommon in humans. Congenital conotruncal defects most commonly reported in veterinary medicine include aorto-pulmonary window and persistent truncus arteriosus. We report a case of an anomalous vessel connecting the ascending aorta to the right pulmonary artery causing left-to-right shunting, left-sided volume overload, and pulmonary overcirculation. Transesophageal echocardiography, cardiac catheterization, and contrast-enhanced computed tomography assisted in the diagnosis and facilitated the surgical correction of the anomalous vessel. The authors hypothesize this defect represents an unusual anomalous vessel connecting the ascending aorta to the right pulmonary artery. PMID- 21640677 TI - Bioprosthesis valve replacement in dogs with congenital tricuspid valve dysplasia: technique and outcome. AB - OBJECTIVE: To describe the surgical technique and report outcome of dogs undergoing bioprosthesis valve replacement for severe tricuspid regurgitation (TR) secondary to congenital tricuspid valve dysplasia (TVD). ANIMALS, MATERIALS AND METHODS: Twelve client-owned dogs (19-43 kg) with TVD underwent tricuspid valve replacement with a bovine pericardial or porcine aortic bioprosthesis with the aid of cardiopulmonary bypass. Anticoagulation with warfarin was maintained for 3 months after surgery and then discontinued. RESULTS: Ten of 12 (83.3%) dogs survived surgery and were discharged from the hospital. Seven dogs were alive with complete resolution of TR for a median period of 48 months (range 1-66 months) after surgery. Two dogs underwent euthanasia because of bioprosthesis failure due to inflammatory pannus at 10 and 13 months after surgery. Two dogs experienced valve thrombosis that was resolved by tissue plasminogen activator. One dog developed suspected endocarditis after surgery that was resolved with antibiotics. Serious cardiac complications included atrial fibrillation and flutter, right-to-left shunt through an uncorrected patent foramen ovale, complete atrioventricular block, and sudden cardiac arrest. Postoperative atrial fibrillation or flutter did not occur in 7 dogs treated prophylactically with oral amiodarone before surgery. CONCLUSIONS: Curative intermediate-term outcomes are possible in dogs undergoing open tricuspid valve replacement with a bioprosthesis. Prosthesis-related complications include inflammatory pannus, thrombosis, and endocarditis. Postoperative atrial fibrillation or flutter can be reduced or prevented by prophylactic preoperative treatment with amiodarone. Several identified complications are avoidable or can be reduced with increased awareness and experience with these techniques. PMID- 21640678 TI - Status of therapeutic gene transfer to treat cardiovascular disease in dogs and cats. AB - Gene therapy is a procedure resulting in the transfer of a gene(s) into an individual's cells to treat a disease, which is designed to produce a protein or functional RNA (the gene product). Although most current gene therapy clinical trials focus on cancer and inherited diseases, multiple studies have evaluated the efficacy of gene therapy to abrogate various forms of heart disease. Indeed, human clinical trials are currently underway. One goal of gene transfer may be to express a functional gene when the endogenous gene is inactive. Alternatively, complex diseases such as end stage heart failure are characterized by a number of abnormalities at the cellular level, many of which can be targeted using gene delivery to alter myocardial protein levels. This review will discuss issues related to gene vector systems, gene delivery strategies and two cardiovascular diseases in dogs successfully treated with therapeutic gene delivery. PMID- 21640679 TI - Population genetic data for 17 Y STR markers from Benghazi (East Libya). AB - The seventeen Y-STR loci included in the AmpFlSTR((r)) YfilerTM PCR Amplification kit (DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS385a/b, DYS438, DYS439, DYS437, DYS448, DYS458, DYS456, DYS635, and Y-GATA-H4) were used to type a sample population of 238 males from eastern Libya (Benghazi region). Of 238 observed haplotypes, 214 were unique (90%) and 24 (10%) were found more than once. The 17 loci gave a discriminating power of 0.999. DYS458 showed the highest diversity as a single-locus marker (0.73). Allelic frequencies and gene diversities for each Y-STR locus were determined. The high haplotype diversity and discrimination capacity (0.996) demonstrate the utility of these loci for human identification in forensic applications. Comparative analysis with Y-STR datasets of relevant populations and submission of the haplotypes to the Y-STR Haplotype Reference Database (YHRD) was undertaken. PMID- 21640680 TI - Medical home disparities between children with public and private insurance. AB - OBJECTIVE: To compare the prevalence of a medical home for children with public versus private insurance and identify components of the medical home that contribute to any differences. METHODS: We performed a secondary data analysis of the 2007 National Survey of Children's Health. A medical home was defined as meeting each of 5 components: 1) usual source of care; 2) personal doctor/nurse; 3) family-centered care; 4) care coordination, if needed; and 5) no problems getting a referral, if needed. We estimated the national prevalence of the medical home and its components for children with public versus private insurance. Comparisons were made using logistic regression, unadjusted and adjusted for sociodemographic factors. RESULTS: A total of 67% of privately insured children met all 5 components of the medical home, compared with only 45% of publicly insured children (P < .001). The gap in medical home prevalence between public and private groups remained significant after controlling for sociodemographic characteristics (public vs private adjusted odds ratio [AOR] 0.82; 95% confidence interval [95% CI] 0.73-0.92). Over 90% of children in both groups reported having a usual source of care and a personal doctor/nurse. Only 58% of publicly insured children reported family-centered care, compared with 76% of privately insured children (P < .001). This difference was significant after adjustment for sociodemographic characteristics (public vs private AOR 0.87; 95% CI 0.77-0.99). CONCLUSIONS: Significant medical home disparities exist between publicly and privately insured children, driven primarily by disparities in family-centered care. Efforts to promote the medical home must recognize and address determinants of family-centered care. PMID- 21640681 TI - Association of externalizing behavior disorder symptoms and injury among fifth graders. AB - OBJECTIVE: Injury is the leading cause of death among American youth, killing more 11-year-olds than all other causes combined. Children with symptoms of externalizing behavior disorders such as attention-deficit/hyperactivity disorder (ADHD) and conduct disorder (CD) may have increased risk. Our aims were to determine: (1) whether increasing symptoms of ADHD and CD associate positively with injuries among a community sample of fifth graders; and (2) whether symptoms of ADHD and CD have a multiplicative rather than additive association with injuries among the sample. METHODS: Data were collected from 4745 fifth graders and their primary caregivers participating in Healthy Passages, a multisite, community-based study of pediatric health risk behaviors and health outcomes. The primary outcome was injury frequency. Primary independent variables were ADHD and CD symptoms. Additional covariates included gender, race/ethnicity, and household income. Ordinal logistic regression examined correlates of injury frequency. The interaction between ADHD and CD symptoms also was examined. RESULTS: In bivariate analyses, the odds of injury increased as ADHD symptoms (odds ratio [OR] 1.29; 95% confidence interval [95% CI] 1.18-1.41) and CD symptoms (OR 1.18; 95% CI 1.07 1.31) increased. However, in multivariate analysis, only ADHD symptoms were significantly associated with injury (OR 1.22; 95% CI 1.10-1.35). There was no statistically significant interaction between ADHD and CD symptoms. CONCLUSIONS: ADHD symptoms are associated with increased odds of injury in fifth graders. Findings have implications for potential injury prevention strategies for mental health practitioners (for example, cognitive training with at-risk youth), pediatricians (ADHD screening), and parents (improved supervision). PMID- 21640682 TI - Annual report on health care for children and youth in the United States: focus on trends in hospital use and quality. AB - OBJECTIVE: The aim of this study was to describe selected trends in hospital inpatient care for children between 2000 and 2007. STUDY DESIGN: Analysis was conducted of administrative data from annual nationwide databases of hospital discharges from the Agency for Healthcare Research and Quality's Healthcare Cost and Utilization Project, along with survey data from a nationally representative random sample of children from the Medical Expenditure Panel Survey. Hospital utilization rates and expenses, risk-adjusted rates of potentially avoidable hospitalization, and safety indicators in the hospital are calculated and tracked with established and downloadable software. RESULTS: The rate of hospital discharges for children aged 15 to 17 years declined significantly, mainly due to fewer maternity-related discharges. The leading principal conditions by age group were similar to the report for 1995 to 2000; however, the rate of admissions for skin infections doubled to 9 per 10,000. Hospital cost per discharge increased by an annual average of 4.5% per year compared with 2.6% annual growth in the gross domestic product deflator. Medicaid is increasingly important relative to private insurance as a payer for hospital care for children. The rate of potentially preventable hospitalizations for both acute and chronic conditions declined substantially (18%, adjusted for age and gender). Several measures of patient safety improved--the rates of postoperative sepsis, iatrogenic pneumothorax, and selected infections due to medical care declined by 14.2%, 17.8%, and 23.5%, respectively. However, the rate of accidental punctures and lacerations and the rate of decubitus ulcer increased by 25.6% and 34.5%, respectively. The trends in safety indicators varied somewhat by age group, income quartile of zip codes, insurance, region, and type of location without a consistent pattern. CONCLUSIONS/IMPLICATIONS: Although teenage pregnancy rates were declining, there was a worsening trend in skin infections. The latter may eventually be impacted by recent publication of new guidelines for treatment by office-based physicians. A gradually increasing role of Medicaid as a payer for hospital care for children will likely put an increasing strain on public resources in advance of the full implementation of the health insurance reforms recently enacted. The decline in potentially avoidable admissions reduces the use of the most expensive resources. For asthma and diabetes, children in the lowest income zip codes had persistently higher rates of admission, but the rate fell by one third during the period. Children in the South and West regions had substantial and significant declines in preventable admissions. Particular indicators of safety were improving, whereas others were worsening. Trends were not the same in all types of hospitals, all regions, and income categories. This is already a rich area for further research on the impact of quality improvement strategies; however, attention is needed to developing more tools to more thoroughly track quality of care for children. PMID- 21640683 TI - Training in social determinants of health in primary care: does it change resident behavior? AB - OBJECTIVE: The aim of this study was to examine the effects of a new social determinants of health curriculum on pediatric interns' attitudes, knowledge, documentation, and clinical practice. METHODS: A nonrandomized mixed-methods study of an educational intervention conducted over a 1-year period was performed. The 2008-2009 pediatric interns (intervention group) participated in a new social determinants of health curriculum; prior year interns were controls. An anonymous online survey at the end of internship to both groups (post-tests) and the beginning of internship to the intervention group (pretest) assessed attitudes and knowledge. Documentation from the electronic medical record of social history questions was audited during the same 3-month period in successive years. Medical-legal partnership (MLP) referrals from both groups were compared. RESULTS: Intervention interns (n = 20) were more comfortable discussing issues (100% vs. 71%; P < .01) and felt more knowledgeable regarding issues (100% vs. 64%; P = .005), community resources (94% vs. 29%; P < .001), and housing (39% vs. 6%; P = .04) than control group interns (n = 18). No differences regarding the importance of social hardships or screening for food security or education issues were found. Knowledge was greater in the intervention group post-test in all domains: benefits (72% vs. 52%), housing (48% vs. 21%), and education (52% vs. 33%; P < .001 for all). Intervention interns were more likely to document each issue (benefits 98% vs. 60%, housing 93% vs. 57%, food 74% vs. 56%; P < .001 for all). The intervention group had a slightly higher rate of referral to MLP, although the difference did not reach statistical significance. CONCLUSION: The educational intervention increased interns' comfort and knowledge of social determinants of health and community resources. Documentation of social questions also increased. PMID- 21640684 TI - Graduating med-peds residents' interest in part-time employment. AB - OBJECTIVE: As part-time work is becoming more popular among the primary care specialties, we examined the demographic descriptors of med-peds residents seeking and finding part-time employment upon completion of residency training. METHODS: As part of the 2006 annual American Academy of Pediatrics (AAP) Graduating Med-Peds Residents Survey, we surveyed the graduating residents of all med-peds programs about their interest in and plans for part-time employment. A total of 199 (60%) of the residents responded. RESULTS: Of the resident respondents applying for nonfellowship jobs, 19% sought part-time positions and 10% actually accepted a part-time position. Female residents were significantly more likely than male residents to apply for part-time jobs (26% vs. 7%, P = .034). Sixty percent of female residents immediately seeking work and 58% of those going on to fellowship reported an interest in arranging a part-time or reduced-hours position at some point in the next 5 years. CONCLUSIONS: Part-time employment among med-peds residents applying for nonfellowship positions after graduation is similar to the current incidence of part-time employment in other fields of primary care. A much higher percentage of med-peds residents are interested in arranging part-time work within 5 years after graduation. This strong interest in part-time work has many implications for the primary care workforce. PMID- 21640685 TI - Hospital care for children in the United States: recent trends and challenges ahead. PMID- 21640686 TI - Dorsally displaced distal radius fractures treated by fixed-angle volar plating: Grip and pronosupination strength recovery. A prospective study. AB - INTRODUCTION: Malunion following open reduction and internal fixation of distal radius fracture threatens wrist function. Fixed-angle palmar plates provide rigid fixation that is stable over time; however, the pronator quadratus sectioning required by the anterior approach entails a risk of pronation strength loss and of distal radioulnar joint destabilization. The present study assessed recovery of grip, pronation and supination strength following such internal fixation. PATIENTS AND METHOD: A prospective study included 26 distal radial fractures with dorsal displacement, osteosynthesized using a fixed-angle palmar plate, in 25 patients (mean age: 47.5 years; range: 17-72 years). Assessment concerned the classical parameters, plus grip and pronosupination strength recovery. RESULTS: At a mean 14 months follow-up (range: 6-30 months), patients had recovered 91% grip strength, 88% pronation strength and 85% supination strength with respect to the healthy side. Complications comprised three cases of malunion, two of reflex sympathetic dystrophy syndrome, and four of post-traumatic carpal tunnel syndrome. DISCUSSION: A study of the literature found 75-95% grip strength recovery following osteosynthesis using fixed-angle plates. Few studies, however, have focused on pronosupination strength, and none reported its evolution following osteosynthesis. CONCLUSION: The present study found no drawbacks associated with a technique which usually involves sectioning the pronator quadratus. Except in case of malunion or joint stiffness, fixed-angle palmar plate osteosynthesis was followed by recovery of grip and pronosupination strength. LEVEL OF EVIDENCE: Level IV: prospective non-randomized, non comparative observational study. PMID- 21640687 TI - Effects of the L/N-type calcium channel antagonist cilnidipine on morning blood pressure control and peripheral edema formation. AB - The L/N-type calcium channel blocker cilnidipine has unique effects including sympathetic nerve suppression and the balanced vasodilatation of arteries and veins that may alleviate morning hypertension (MHT) or peripheral edema caused by calcium channel antagonists. We used ambulatory blood pressure monitoring (ABPM) and a unique peripheral edema measurement to evaluate the effect of morning and bedtime cilnidipine in patients with MHT. Forty-three patients with MHT (60 +/- 12 years) were randomly assigned to a morning or bedtime cilnidipine (10-20 mg/day). MHT was defined as a mean systolic blood pressure (SBP) >= 135 mm Hg by ABPM within 2 hours after awaking. After 3 months, greater SBP reductions were observed in the bedtime administration group (versus the morning administration group) at 3:30-6:00 AM (-24 +/- 20 mm Hg vs. -10 +/- 4 mm Hg; P < .05) and at 6:30-9:00 AM (-26 +/- 15 mm Hg vs. -14 +/- 17 mm Hg; P < .05). Although physical examinations showed leg edema in 16% of the patients, quantitative evaluations did not reveal significant volume gains. Cilnidipine had a greater effect on MHT, without causing significant leg edema, when administered at bedtime. PMID- 21640688 TI - Translation of hypertension treatment guidelines into practice: a review of implementation. AB - Compared with the history of national guideline development, the science attached to implementation of guidelines is relatively new. Effectiveness of a highly evidence-based guideline, such as the 8th Joint National Committee recommendations on the treatment of high blood pressure, depends on successful translation into clinical practice. Implementation relies on several steps: clear and executable guideline language, audit and feedback attached to education of practitioners charged with carrying out the guidelines, team-based care delivery, credibility of blood pressure measurement, and measures to address therapeutic inertia and medication adherence. An evolving role of the electronic health record and patient empowerment are developments that will further promote implementation of the hypertension guideline. Further research will be needed to assess the efficacy and cost effectiveness of various implementation tools and strategies. PMID- 21640689 TI - The effect of eprosartan on reflex sympathetic activation in sodium restricted patients with essential hypertension. AB - AT(1) receptor antagonists possess sympathoinhibitory effects in animal experiments, but in human studies the results are conflicting. We tested the hypothesis that very short-term treatment with the AT(1) receptor antagonist eprosartan inhibits reflex activation of the sympathetic nervous system in sodium restricted patients with essential hypertension. The effect of eprosartan on urinary sodium and lithium excretion, heart rate, blood pressure, and vasoactive hormones was measured during reflex activation of the sympathetic nervous system by a cold pressor test and by a sodium nitroprusside induced 10 mm Hg reduction of the mean arterial pressure. It was a randomized, placebo-controlled, double blinded, crossover study in 14 patients with essential hypertension. Glomerular filtration rate and renal tubular function were determined with continuous infusion clearance technique and vasoactive hormones with radioimmunoassays. Eprosartan had no effect on the increases in heart rate and plasma levels of noradrenaline during reflex activation of the sympathetic nervous system. However, eprosartan significantly decreased in fractional excretions of sodium (mean +/- SD) (0.23 +/- 0.22%) and lithium (3.1 +/- 1.7%) during the sodium nitroprusside infusion, compared to placebo. Very short-term eprosartan treatment does not seem to have any sympathoinhibitory effects in sodium restricted patients with essential hypertension. PMID- 21640690 TI - Standardized medical terminology for cardiac computed tomography: a report of the Society of Cardiovascular Computed Tomography. AB - Since the emergence of cardiac computed tomography (CT) at the turn of the 21st century, there has been an exponential growth in research and clinical development of the technique, with contributions from investigators and clinicians from varied backgrounds: physics and engineering, informatics, cardiology, and radiology. However, terminology for the field is not unified. As a consequence, there are multiple abbreviations for some terms, multiple terms for some concepts, and some concepts that lack clear definitions and/or usage. In an effort to aid the work of all those who seek to contribute to the literature, clinical practice, and investigation of the field, the Society of Cardiovascular Computed Tomography sets forth a standard set of medical terms commonly used in clinical and investigative practice of cardiac CT. PMID- 21640691 TI - Endoscopic submucosal dissection of scar-embedded rectal polyps: a prospective study (Esd in scar-embedded rectal polyps). AB - BACKGROUND & AIMS: Endoscopic submucosal dissection (ESD) was developed for en bloc resection of superficial neoplasm of the digestive tract. We evaluated feasibility and safety of ESD, as a salvage therapy of large refractory rectal polyps, in a tertiary care setting. METHODS: We prospectively enrolled in the present study and treated by ESD 11 consecutive patients with rectal polyps (median diameter 3.5 cm; range 2-5 cm), who had previously undergone several attempts of endoscopic resection and not suitable for further standard endoscopic treatment. The ESD was carried out with a standard needle knife. Follow up examinations were scheduled at 3, 6, 12 and 24 months. RESULTS: We achieved apparently complete resection of polyps in 10/11 patients. In one patient ESD was interrupted and the pathology of the resected fragment showed deep submucosal infiltration; this patient underwent surgery. Deep and lateral margins were shown to be free of neoplasm (radical resection) in six out of 11 patients. However all the 10 patients with apparently complete resection were free of recurrence after a mean follow up of 19.2 months (12-24). A T1 adenocarcinoma was radically resected by ESD, with no recurrence. We recorded 2 cases of subcutaneous emphysema, both treated conservatively. CONCLUSIONS: Radical resection is difficult to be achieved by ESD in patients with rectal scar-embedded polyps. Nevertheless ESD may be proposed as a definitive treatment of selected patients with refractory polyps, avoiding surgery in the majority of them. PMID- 21640692 TI - Re: Prospective evaluation of operating characteristics of prostate cancer detection biomarkers. PMID- 21640693 TI - Re: Magnetic resonance imaging guided prostate biopsy in men with repeat negative biopsies and increased prostate specific antigen. PMID- 21640694 TI - Re: Prostate cancer antigen 3 score accurately predicts tumour volume and might help in selecting prostate cancer patients for active surveillance. PMID- 21640695 TI - Re: Predictors of androgen deprivation therapy efficacy combined with prostatic irradiation: the central role of tumor stage and radiation dose. PMID- 21640696 TI - Re: Posterior rhabdosphincter reconstruction during robotic assisted radical prostatectomy: results from a phase II randomized clinical trial. PMID- 21640697 TI - Re: Comparison of cold and warm ischemia during partial nephrectomy in 660 solitary kidneys reveals predominant role of nonmodifiable factors in determining ultimate renal function. PMID- 21640698 TI - An inexpensive high-throughput nuclear magnetic resonance tube cleaning apparatus. AB - Large-scale nuclear magnetic resonance (NMR) tube cleaning is currently a bottleneck in high-throughput NMR ligand affinity screens. Expensive alternatives include discarding the NMR tubes after a single use (~US $2-$8/tube), using commercial NMR tube cleaners (~$15,000), and abandoning NMR tubes for flow probe technology (~$75,000). Instead, we describe a relatively inexpensive (~$400) and easily constructed apparatus that can clean 180 NMR tubes per hour while using a modest amount of solvent. The application of this apparatus significantly shortens the time to recycle NMR tubes while avoiding cross-contamination and tube damage. PMID- 21640699 TI - Efficient removal of detergents from proteins and peptides in a spin column format. AB - Detergents are commonly used in protein-chemistry protocols and may be necessary for protein extraction, solubilization, and denaturation; however, their presence interferes with many downstream analysis techniques, including mass spectrometry (MS). To enable downstream analysis, it is critical to remove unbound detergents from protein and peptide samples. In this study, we describe a high-performance resin that offers exceptional detergent removal for proteins and peptides. When used in a spin column format, this resin dramatically improves protein and peptide MS results by more than 95% removal of 1-5% detergents, including sodium dodecyl sulfate (SDS), sodium deoxycholate, Chaps, Triton X-100, Triton X-114, NP 40, Brij-35, octyl glucoside, octyl thioglucoside, and lauryl maltoside, with high recovery of proteins and peptides. Postcolumn liquid chromatography-tandem MS (LC-MS/MS) analysis of trypsin digests of bovine serum albumin (BSA) and HeLa cell lysate revealed excellent sequence coverage, indicating successful removal of detergent from the peptides. Matrix-assisted laser desorption/ionization (MALDI)-MS analysis of unprocessed and processed samples further confirmed efficient removal of detergents. The advantages of this method include speed (<15min), efficient detergent removal, and high recovery of proteins and peptides. PMID- 21640701 TI - Role of calcium in the conformational dynamics of factor XIII activation examined by hydrogen-deuterium exchange coupled with MALDI-TOF MS. AB - Factor XIII catalyzes formation of gamma-glutamyl-epsilon-lysyl crosslinks within fibrin clots. FXIII A(2) can be activated proteolytically with thrombin and low mM Ca(2+) or nonproteolytically with high monovalent/divalent cations along with low mM Ca(2+). Physiologically, FXIII A(2) is poised to respond to transient influxes of Ca(2+) in a Na(+) containing environment. A successful strategy to monitor FXIII conformational events is hydrogen-deuterium exchange (HDX) coupled with mass spectrometry. FXIII A(2) was examined in the presence of different cations (Ca(2+), Mg(2+), Ba(2+), Cu(2+), Na(+), TMAC(+), and EDA(2+)) ranging from 1 to 2mM, physiological Ca(2+) concentration, to 50-500mM for nonproteolytic activation. Increases in FXIII solvent exposure could already be observed at 1mM Ca(2+) for the dimer interface, the catalytic site, and glutamine substrate regions. By contrast, solvent protection was observed at the secondary cleavage site. These events occurred even though 1mM Ca(2+) is insufficient for FXIII activation. The metals 1mM Mg(2+), 1mM Ba(2+), and 1mM Cu(2+) each led to conformational changes, many in the same FXIII regions as Ca(2+). FXIII could also be activated nonproteolytically with 500mM tetramethylammonium chloride (TMAC(+)) and 500mM ethylenediamine (EDA(2+)), both with 2mM Ca(2+). These different HDX studies help reveal the first FXIII segments that respond to physiological Ca(2+) levels. PMID- 21640700 TI - Cellular magnesium homeostasis. AB - Magnesium, the second most abundant cellular cation after potassium, is essential to regulate numerous cellular functions and enzymes, including ion channels, metabolic cycles, and signaling pathways, as attested by more than 1000 entries in the literature. Despite significant recent progress, however, our understanding of how cells regulate Mg(2+) homeostasis and transport still remains incomplete. For example, the occurrence of major fluxes of Mg(2+) in either direction across the plasma membrane of mammalian cells following metabolic or hormonal stimuli has been extensively documented. Yet, the mechanisms ultimately responsible for magnesium extrusion across the cell membrane have not been cloned. Even less is known about the regulation in cellular organelles. The present review is aimed at providing the reader with a comprehensive and up-to-date understanding of the mechanisms enacted by eukaryotic cells to regulate cellular Mg(2+) homeostasis and how these mechanisms are altered under specific pathological conditions. PMID- 21640702 TI - Interaction of aryl hydrocarbon receptor and NF-kappaB subunit RelB in breast cancer is associated with interleukin-8 overexpression. AB - The aryl hydrocarbon receptor (AhR) has been best known for its role in mediating the toxicity of dioxin. Here we show that AhR overexpression is found among estrogen receptor (ER)alpha-negative human breast tumors and that its overexpression is positively correlated to that of the NF-kappaB subunit RelB and Interleukin (IL)-8. Increased DNA binding activity of the AhR and RelB is coupled to IL-8 overexpression in primary breast cancer tissue, which was also supported by in situ hybridization. Activation of AhR in vitro by 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) induced IL-8 expression in MDA-MB 436 and MCF 7 cells in an AhR and RelB dependent manner. Consistently, downregulation of RelB or AhR by small interfering RNAs (siRNA) decreased the level of IL-8 but increased expression of ERalpha in vitro in MCF-7 cells. Our results strongly suggest that RelB and AhR have a critical role in the regulation of IL-8 and reveal a supportive role of RelB and AhR in the anti-apoptotic response in human breast cancer cells. AhR and RelB may present a novel therapeutic target for inflammatory driven breast carcinogenesis and tumor progression. Overexpression of pro-survival factors AhR and RelB may explain the process of the development of environmentally-induced type of breast cancers. PMID- 21640703 TI - Molecular diversity of the two sugar-binding sites of the beta-trefoil lectin HA33/C (HA1) from Clostridium botulinum type C neurotoxin. AB - A critical role in internalizing the Clostridium botulinum neurotoxin into gastrointestinal cells is played by nontoxic components complexed with the toxin. One of the components, a beta-trefoil lectin has been known as HA33 or HA1. The HA33 from C. botulinum type A (HA33/A) has been predicted to have a single sugar binding site, while type C HA33 (HA33/C) has two sites. Here we constructed HA33/C mutants and evaluated the binding capacities of the individual sites through mucin-assay and isothermal titration calorimetry. The mutant W176A (site I knockout) had a K(d) value of 31.5mM for galactose (Gal) and 61.3mM for N acetylgalactosamine (GalNAc), while the K(d) value for N-acetylneuraminic acid (Neu5Ac) was too high to be determined. In contrast, the double mutant N278A/Q279A (site II knockout) had a K(d) value of 11.8mM for Neu5Ac. We also determined the crystal structures of wild-type and the F179I mutant in complex with GalNAc at site II. The results suggest that site I of HA33/C is quite unique in that it mainly recognizes Neu5Ac, and site II seems less important for the lectin specificity. The architectures and the properties of the sugar-binding sites of HA33/C and HA33/A were shown to be drastically different. PMID- 21640704 TI - Effects of 3T3 adipocytes on interleukin-6 expression and insulin signaling in L6 skeletal muscle cells. AB - OBJECTIVE: Central adiposity and inflammation play key roles in the development of insulin resistance through the effects of pro-inflammatory adipokines such as IL-6, but the effect of infiltrating adipocytes in skeletal muscle tissues is not known. Communications between muscle cells and fat cells may contribute to the inflammatory response associated with insulin resistance. METHODS: In this study we used a co-culture system of skeletal muscle (L6) and adipocyte (3T3-L1) cell lines to study expression of the inflammatory cytokine IL-6 and changes in insulin signaling. This model could mimic the adipocytes infiltrating myocytes that is commonly seen in obese patients. RESULTS: When plated alone the L6 cells express IL-6 mRNA and secrete IL-6 protein, both of which are increased when the cells are challenged with the bacterial lipopolysaccharide (LPS). In contrast, the 3T3-L1 cells had very little expression of IL-6 mRNA or protein. Co-culture of 3T3-L1 pre-adipocytes with L6 cells, at a density ratio of 1:10, respectively, increased IL-6 expression significantly and decreased insulin-stimulated Akt phosphorylation. To examine the role of IL-6 in insulin sensitivity we incubated the L6 cells with IL-6. A brief challenge of L6 cells with IL-6 enhanced insulin stimulated Akt phosphorylation. In contrast, incubation of the L6 cells with IL-6 for 96h markedly decreased insulin-stimulated Akt phosphorylation. CONCLUSION: The enhanced IL-6 mRNA expression and IL-6 release in L6 myocytes co-cultured with 3T3-L1 cells indicate an important interaction between adipocytes and myocytes. This observation may shed some light on the long-standing enigma of obesity-induced insulin resistance where infiltration of the skeletal muscle by preadipocytes/adipocytes is evident. PMID- 21640705 TI - Ribonucleotide reductase subunit p53R2 regulates mitochondria homeostasis and function in KB and PC-3 cancer cells. AB - Ribonucleotide reductase (RR) is a rate-limiting enzyme that catalyzes de novo conversion of ribonucleotide 5'-diphosphates to the corresponding 2' deoxynucleotide, essential for DNA synthesis and replication. The mutations or knockout of RR small subunit, p53R2, results in the depletion of mitochondrial DNA (mtDNA) in human, implying that p53R2 might play a critical role for maintaining mitochondrial homeostasis. In this study, siRNA against p53R2 knockdown approach is utilized to examine the impact of p53R2 depletion on mitochondria and to derive underlying mechanism in KB and PC-3 cancer cells. Our results reveal that the p53R2 expression not only positively correlates with mtDNA content, but also partakes in the proper mitochondria function, such as ATP synthesis, cytochrome c oxidase activity and membrane potential maintenance. Furthermore, overexpression of p53R2 reduces intracellular ROS and protects the mitochondrial membrane potential against oxidative stress. Unexpectedly, knockdown of p53R2 has a modest, if any, effect on mitochondrial and total cellular dNTP pools. Taken together, our study provides functional evidence that mitochondria is one of p53R2-targeted organelles and suggests an unexpected function of p53R2, which is beyond known RR function on dNTP synthesis, in mitochondrial homeostatic control. PMID- 21640706 TI - Role of aromatic stack pairing at the catalytic site of gelonin protein. AB - Aromatic-aromatic interactions play an important role in the enzyme-substrate recognition mechanism and in stabilization of proteins. Gelonin--a ribosome inactivating protein (RIP) from the plant Gelonium multiflorum--belongs to type-I RIPs and shows N-glycosylation activity which has been used as a model to explain the role of aromatic-aromatic stack pairing in RIPs. RIPs have a different substrate binding site and catalytic site. Role of tyrosine residues at the binding site has already been known but the role of tyrosine residues at catalytic site is still unclear. In this study, the role of tyrosine-adenine tyrosine aromatic stack pairing at the catalytic site was studied by in silico mutation studies using molecular dynamic simulations. Through this study we report that, despite the fact that aromatic stack pairing aids in recognition of adenine at binding site, both the tyrosine residues of stack pairing play a crucial role in the stabilization of adenine at catalytic site. In the absence of both the tyrosine residues, adenine was unstable at catalytic site that results in the inhibition of N-glycosylation activity of gelonin protein. Hence, this study highlights the importance of pi-pi stack pairing in the N-glycosidic activity of gelonin by determining its role in stabilizing adenine at catalytic site. PMID- 21640707 TI - Activation of peroxisome proliferator-activated receptor-alpha (PPARalpha) suppresses postprandial lipidemia through fatty acid oxidation in enterocytes. AB - Activation of peroxisome proliferator-activated receptor (PPAR)-alpha which regulates lipid metabolism in peripheral tissues such as the liver and skeletal muscle, decreases circulating lipid levels, thus improving hyperlipidemia under fasting conditions. Recently, postprandial serum lipid levels have been found to correlate more closely to cardiovascular diseases than fasting levels, although fasting hyperlipidemia is considered an important risk of cardiovascular diseases. However, the effect of PPARalpha activation on postprandial lipidemia has not been clarified. In this study, we examined the effects of PPARalpha activation in enterocytes on lipid secretion and postprandial lipidemia. In Caco 2 enterocytes, bezafibrate, a potent PPARalpha agonist, increased mRNA expression levels of fatty acid oxidation-related genes, such as acyl-CoA oxidase, carnitine palmitoyl transferase, and acyl-CoA synthase, and oxygen consumption rate (OCR) and suppressed secretion levels of both triglycerides and apolipoprotein B into the basolateral side. In vivo experiments revealed that feeding high-fat-diet containing bezafibrate increased mRNA expression levels of fatty acid oxidation related genes and production of CO(2) and acid soluble metabolites in enterocytes. Moreover, bezafibrate treatment suppressed postprandial lipidemia after oral administration of olive oil to the mice. These findings indicate that PPARalpha activation suppresses postprandial lipidemia through enhancement of fatty acid oxidation in enterocytes, suggesting that intestinal lipid metabolism regulated by PPARalpha activity is a novel target of PPARalpha agonist for decreasing circulating levels of lipids under postprandial conditions. PMID- 21640708 TI - Mechanism of activation of human c-KIT kinase by internal tandem duplications of the juxtamembrane domain and point mutations at aspartic acid 816. AB - The proto-oncogene c-KIT receptor has been implicated as an essential component in the activation of leukemic cells. The internal tandem duplication (ITD) of c KIT has also been identified as a predominant cause of acute myeloid leukemia (AML), although its role in the activation process is still unclear. To investigate the biological mechanisms of c-KIT activation, we generated a c-KIT receptor bearing two different immunological tags, HA and Flag tags. In this study, we demonstrated that the mutant (Mt)-ITD and Asp816 (D816Y) c-KIT receptors spontaneously formed dimers and that these Mt-ITD forms of c-KIT displayed high levels of phosphorylation and increased cellular tyrosine phosphorylation. The amount of wild-type homodimers increased following the addition of the c-KIT ligand, while the level of mutant homodimers was less affected by the addition of the c-KIT ligand. Furthermore, we demonstrated that Mt-ITD and activating point mutations of D816Y induced constitutive activation of c-KIT kinase in the absence of ligand in COS-1 cells. These data suggest a novel mechanism for the regulation of cell growth autonomy. Overall, our study suggests that c-KIT activation might have significant effects on hematopoietic cells and might help to improve our understanding of the pathogenesis of systemic mast cell disease, gastrointestinal stromal tumors and AML and potentially lead to the development of novel therapeutic approaches. PMID- 21640709 TI - Stretch induced endothelin-1 secretion by adult rat astrocytes involves calcium influx via stretch-activated ion channels (SACs). AB - The expression of endothelins (ETs) and ET-receptors is often upregulated in brain pathology. ET-1, a potent vasoconstrictor, also inhibits the expression of astrocyte glutamate transporters and is mitogenic for astrocytes, glioma cells, neurons, and brain capillary endothelia. We have previously shown that mechanical stress stimulates ET-1 production by adult rat astrocytes. We now show in adult astrocytes that ET-1 production is driven by calcium influx through stretch activated ion channels (SACs) and the ET-1 production correlates with cell proliferation. Mechanical stimulation using biaxial stretch (<20%) of a rubber substrate increased ET-1 secretion, and 4 MUM GsMTx-4 (a specific inhibitor of SACs) inhibited secretion by 30%. GsMTx-4 did not alter basal ET-1 levels in the absence of stretch. Decreasing the calcium influx by lowering extracellular calcium also inhibited stretch-induced ET-1 secretion without effecting ET-1 secretion in unstretched controls. Furthermore, inhibiting SACs with the less specific inhibitor streptomycin also inhibited stretch-induced ET-1 secretion. The data can be explained with a simple model in which ET-1 secretion depends on an internal Ca(2+) threshold. This coupling of mechanical stress to the astrocyte endothelin system through SACs has treatment implications, since all pathology deforms the surrounding parenchyma. PMID- 21640710 TI - MicroRNAs expression in ox-LDL treated HUVECs: MiR-365 modulates apoptosis and Bcl-2 expression. AB - Endothelial cells (ECs) apoptosis induced by oxidized low-density lipoprotein (ox LDL) is thought to play a critical role in atherosclerosis. MicroRNAs (miRNAs) are a class of noncoding RNAs that posttranscriptionally regulate the expression of genes involved in diverse cell functions, including differentiation, growth, proliferation, and apoptosis. However, whether miRNAs are associated with ox-LDL induced apoptosis and their effect on ECs is still unknown. Therefore, this study evaluated potential miRNAs and their involvement in ECs apoptosis in response to ox-LDL stimulation. Microarray and qRT-PCR analysis performed on human umbilical vein endothelial cells (HUVECs) exposed to ox-LDL identified 15 differentially expressed (4 up- and 11 down-regulated) miRNAs. Web-based query tools were utilized to predict the target genes of the differentially expressed miRNAs, and the potential target genes were classified into different function categories with the gene ontology (GO) term and KEGG pathway annotation. In particular, bioinformatics analysis suggested that anti-apoptotic protein B-cell CLL/lymphoma 2 (Bcl-2) is a target gene of miR-365, an apoptomir up-regulated by ox-LDL stimulation in HUVECs. We further showed that transfection of miR-365 inhibitor partly restored Bcl-2 expression at both mRNA and protein levels, leading to a reduction of ox-LDL-mediated apoptosis in HUVECs. Taken together, our findings indicate that miRNAs participate in ox-LDL-mediated apoptosis in HUVECs. MiR-365 potentiates ox-LDL-induced ECs apoptosis by regulating the expression of Bcl-2, suggesting potential novel therapeutic targets for atherosclerosis. PMID- 21640711 TI - Mitochondrial dysfunction induced by knockdown of mortalin is rescued by Parkin. AB - Mutations in the parkin gene are the most common cause of autosomal recessive Parkinson's disease (PD). As an E3-ubiquitin ligase, Parkin is associated with mitochondrial dynamics and mitophagy. Mortalin, a molecular chaperone, is located primarily in mitochondria, where it functions to maintain mitochondrial homeostasis and antagonize oxidative stress injury. A reduced expression level of mortalin has been observed in the affected brain regions of PD patients. Mortalin also interacts with a variety of PD-related proteins and plays an indispensible role in helping native protein refolding and importing proteins into the mitochondrial matrix. Thus, the main aims of the present study were to investigate mitochondrial dysfunction induced by knockdown of mortalin and to test whether Parkin overexpression could rescue this effect. We found that lentivirus-mediated knockdown of mortalin in HeLa cells resulted in a collapse of mitochondrial membrane potential, an abnormal accumulation of reactive oxygen species and apparent alterations in mitochondrial morphology under H(2)O(2) induced stress conditions. Remarkably, Parkin overexpression rescued these mitochondrial abnormalities. In HeLa cells expressing Parkin, co immunoprecipitation of endogenous mortalin and wild-type Parkin was detected when they were treated with carbonyl cyanide 3-chlorophenylhydrazone (CCCP). In conclusion, we indicate that the relatively decreased mortalin expression level and its impaired interaction with Parkin could affect its roles in mitochondrial function. PMID- 21640712 TI - Study of the docking-dependent PLK1 phosphorylation of the CDC25B phosphatase. AB - CDC25 (A, B and C) phosphatases control cell cycle progression through the timely dephosphorylation and activation of cyclin-dependent kinases (CDK). At mitosis the CDC25B phosphatase activity is dependent on its phosphorylation by multiple kinases impinging on its localisation, stability and catalytic activity. Here we report that prior phosphorylation of CDC25B by CDK1 enhances its substrate properties for PLK1 in vitro, and we also show that phosphorylated S50 serves as a docking site for PLK1. Using a sophisticated strategy based on the sequential phosphorylation of CDC25B with (16)O and (18)O ATP prior to nanoLC-MS/MS analysis we identified 13 sites phosphorylated by PLK1. This study illustrates the complexity of the phosphorylation pattern and of the subsequent regulation of CDC25B activity. PMID- 21640713 TI - CD3+CD4-CD8- (double negative) T cells: saviours or villains of the immune response? AB - Recent studies have shown that T cells are not just the latecomers in inflammation but might also play a key role in the early phase of this response. In this context, a number of T cell subsets including NKT cells, mucosal associated invariant T cells and gamma/delta T cells have been shown, together with classical innate immune cells, to contribute significantly to the development and establishment of acute and chronic inflammatory diseases. In this commentary we will focus our attention on a somewhat neglected class of T cells called CD3(+)CD4(-)CD8(-) double negative T cells and on their role in inflammation and autoimmunity. We will summarize the most recent views on their origin at the thymic and peripheral levels as well as their tissue localization in immune and non-lymphoid organs. We will then outline their potential pathogenic role in autoimmunity as well as their homeostatic role in suppressing excessive immune responses deleterious to the host. Finally, we will discuss the potential therapeutic benefits or disadvantages of targeting CD3(+)CD4(-)CD8(-) double negative T cells for the treatment of autoimmune disease. We hope that this overview will shed some light on the function of these immune cells and attract the interest of investigators aiming at the design of novel therapeutic approaches for the treatment of autoimmune and inflammatory conditions. PMID- 21640714 TI - Aliskiren affects fatty-acid uptake and lipid-related genes in rodent and human cardiomyocytes. AB - PURPOSE: We investigated whether the direct renin inhibitor aliskiren can affect metabolism in cardiomyocytes from rat, mouse and human sources. METHODS AND RESULTS: At 10-50 MUmol/L, aliskiren significantly increased medium-chain-fatty acid uptake in primary-cultured neonatal-rat and HL-1 adult-mouse-derived cardiomyocytes (BODIPY-induced fluorescence intensity). The fatty-acid transporter CD-36 was correspondingly translocated to, but the glucose transporter Glut-4 away from, the sarcoplasmic reticulum/plasma membrane, in primary-cultured neonatal-rat (CD-36, Glut-4) and adult-human (CD-36) cardiomyocytes (confocal immunocytochemistry). Immunoblotting showed that aliskiren induced phosphorylation of ERK1/2 in cardiomyocytes from all three sources; responses were dose- and time-dependent, unaffected by renin treatment, and did not cause alterations in expression of (P)R or Igf2/M6P receptors. Microarray analysis of the complete genome of aliskiren-treated neonatal-rat cardiomyocytes, with RT-qPCR and immunoblot confirmation assays in rat and human primary cardiomyocytes, showed that aliskiren up-regulated mRNA and increased protein expression of several enzymes important in lipid and glucose metabolism and in cholesterol biosynthesis. Cardiomyocyte cell-cycle and viability were unaffected by aliskiren. CONCLUSIONS: Aliskiren can induce changes in fatty-acid and glucose uptake and expression of key enzymes of lipid and cholesterol metabolism, which are not associated with increased expression of (P)R or Igf2/M6P receptors, in cultured cardiomyocytes. PMID- 21640715 TI - The cytoskeleton plays a modulatory role in the association between STIM1 and the Ca2+ channel subunits Orai1 and TRPC1. AB - Store-operated Ca(2+) entry (SOCE) is a major pathway for Ca(2+) influx in non excitable cells. Recent studies favour a conformational coupling mechanism between the endoplasmic reticulum (ER) Ca(2+) sensor STIM1 and Ca(2+) permeable channels in the plasma membrane to explain SOCE. Previous studies have reported a role for the cytoskeleton modulating the activation of SOCE; therefore, here we have investigated whether the interaction between STIM1 and the Ca(2+) permeable channels is modulated by the actin or microtubular network. In HEK-293 cells, treatment with the microtubular disrupter colchicine enhanced both the activation of SOCE and the association between STIM1 and Orai1 or TRPC1 induced by thapsigargin (TG). Conversely, stabilization of the microtubules by paclitaxel attenuated TG-evoked activation of SOCE and the interaction between STIM1 and the Ca(2+) channels Orai1 and TRPC1, altogether suggesting that the microtubules act as a negative regulator of SOCE. Stabilization of the cortical actin filament layer results in inhibition of TG-evoked both association between STIM1, Orai1 and TRPC1 and SOCE. Interestingly, disruption of the actin filament network by cytochalasin D did not significantly modify TG-evoked association between STIM1 and Orai1 or TRPC1 but enhanced TG-stimulated SOCE. Finally, inhibition of calmodulin by calmidazolium enhances TG-evoked SOCE and disruption of the actin cytoskeleton results in inhibition of TG-evoked association of calmodulin with Orai1 and TRPC1. Thus, we demonstrate that the cytoskeleton plays an essential role in the regulation of SOCE through the modulation of the interaction between their main molecular components. PMID- 21640716 TI - Nicotinic acetylcholine receptor-mediated mechanisms in lung cancer. AB - Despite the known adverse health effects associated with tobacco use, over 45 million adults in the United States smoke. Cigarette smoking is the major etiologic factor associated with lung cancer. Cigarettes contain thousands of toxic chemicals, many of which are carcinogenic. Nicotine contributes directly to lung carcinogenesis through the activation of nicotinic acetylcholine receptors (nAChRs). nAChRs are ligand-gated ion channels, expressed in both normal and lung cancer cells, which mediate the proliferative, pro-survival, angiogenic, and metastatic effects of nicotine and its nitrosamine derivatives. The underlying molecular mechanisms involve increases in intracellular calcium levels and activation of cancer signal transduction pathways. In addition, acetylcholine (ACh) acts as an autocrine or paracrine growth factor in lung cancer. Other neurotransmitters and neuropeptides also activate similar growth loops. Recent genetic studies further support a role for nAChRs in the development of lung cancer. Several nAChR antagonists have been shown to inhibit lung cancer growth, suggesting that nAChRs may serve as valuable targets for biomarker-guided lung cancer interventions. PMID- 21640718 TI - Dual epidermal growth factor receptor (EGFR)/insulin-like growth factor-1 receptor (IGF-1R) inhibitor: a novel approach for overcoming resistance in anticancer treatment. AB - Small molecule inhibitors of epidermal growth factor receptors (EGFR) have been found to show a good initial response in cancer patients but during the course of treatment, patients develop resistance after a few weeks of time. Development of secondary mutations or over-activation of insulin like growth factor (IGF-1R) pathway are a few of the several mechanisms proposed to explain the resistance. To study the effect of dual inhibition of EGFR and IGF-1R in overcoming the resistance, three strategies were envisaged and are reported in this manuscript: 1) a virtual predictive tumor model, 2) in vitro experimental data using a combination of EGFR and IGF-1R inhibitors and 3) in vitro experimental data using in house dual inhibitors. Findings reported in this manuscript suggest that simultaneous inhibition of IGF-1R and EGFR either by combination of two inhibitors or by dual kinase inhibitors is more efficacious compared to single agents. In vitro cell based experiments conducted using epidermoid cancer cell line, A431 and an EGFR mutant cell line, H1975 along with virtual predictions reported here suggests that dual inhibition of EGFR and IGF-1R is a viable approach to overcome EGFR resistance. PMID- 21640717 TI - Ectopic expression of Nkx2.5 suppresses the formation of the sinoatrial node in mice. AB - The sinoatrial node (SAN), functionally known as the pacemaker, regulates the cardiac rhythm or heartbeat. Several genes are expressed in the developing SAN and form a genetic network regulating the fate of the SAN cells. The short stature homeobox gene Shox2 is an important player in the SAN genetic network by regulating the expression of different cardiac conduction molecular markers including the early cardiac differentiation marker Nkx2.5. Here we report that the expression patterns of Shox2 and Nkx2.5 are mutually exclusive from the earliest stages of the venous pole and the SAN formation. We show that tissue specific ectopic expression of Shox2 in the developing mouse heart downregulates the expression of Nkx2.5 and causes cardiac malformations; however, it is not sufficient to induce a SAN cell fate switch in the working myocardium. On the other hand, tissue specific overexpression of Nkx2.5 in the heart leads to severe hypoplasia of the SAN and the venous valves, dis-regulation of the SAN genetic network, and change of the SAN cell fate into working myocardium, and causes embryonic lethality, recapitulating the phenotypes including bradycardia observed in Shox2(-/-) mutants. These results indicate that Nkx2.5 activity is detrimental to the normal formation of the SAN. Taken together, our results demonstrate that Shox2 downregulation of Nkx2.5 is essential for the proper development of the SAN and that Shox2 functions to shield the SAN from becoming working myocardium by acting upstream of Nkx2.5. PMID- 21640719 TI - HMGB1 translocation and expression is caused by warm ischemia reperfusion injury, but not by partial hepatectomy in rats. AB - Mechanical injury or ischemia/reperfusion (I/R) injury induces high mobility of group box 1 (HMGB1) translocation and release. However, the surgical procedure itself can initiate pathophysiologic processes causing damage to the respective organ. A liver resection, as an example, leads to portal hyperperfusion injury of the remnant liver. Therefore, we aimed to elucidate the impact of different hepatic surgical injury models on cellular localization and expression of HMGB1. Focal warm I/R injury was induced by clamping the vascular blood supply to the median and left lateral liver lobes for 90 min followed by 0.5 h, 6 h and 24 h reperfusion, as reported previously. Liver injury by PH was induced by subjecting rats to 30%, 70% or 90% partial hepatectomy (PH) followed by a 24 h observation period. Additional 12 rats were subjected to 90% PH and sacrificed at 1 h and 6 h to investigate the expression and release pattern of HMGB1. Elevation of serum liver enzymes indicating hepatic injury peaked at 6 h and recovered thereafter in models, warm I/R injury and PH. Liver injury was confirmed by liver histology. HMGB1 was translocated from the nucleus to the cytoplasm in livers subjected to warm I/R; but not in livers subjected to PH. Both protein and mRNA expression of HMGB1 were significantly up-regulated in livers subjected to warm I/R. In contrast, neither 30% PH, 70% PH nor 90% PH caused an elevation of hepatic HMGB1 mRNA and protein expression. High serum levels of HMGB1 (30 ng/ml) were measured at 0.5 h reperfusion period after warm I/R, much lower levels thereafter (<5 ng/ml). Similar low serum levels were measured at all time points after 90% PH. Subsequently expression levels of TNF-a should be changed to tumor necrosis factor-alpha (TNF-alpha) reached a peak (26-fold elevation) at 6 h and decreased down to 5-fold at 24 h after warm I/R. TNF-alpha expression levels after PH never exceeded a 5-fold elevation. In conclusion, HMGB1 translocation and expression depends on the type of liver injury as it is induced by ischemia, but not by liver resection/hyperperfusion. These results suggest that HMGB1 may be used as molecular marker to visualize ischemic damage. Mechanic injury in hepatic surgery is associated with focal warm ischemia, and thereby HMGB1 translocation reflects surgical quality in experimental PH. Expression of hepatic TNF-alpha follows the kinetic pattern of HMGB1, pointing to a muss less pronounced inflammatory response after successful PH compared to warm I/R injury. PMID- 21640720 TI - Delta-aminolevulinic dehydratase is a proteasome interacting protein. AB - The proteasome interacts with a large number of proteins which regulate specific cellular functions. The focus of this study is to examine the proteasome interaction with Delta-aminolevulinate dehydratase (ALAD). ALAD is involved in the heme biosynthesis pathway and was co-isolated, with the 20S proteasome using several chromatographic purification steps. The MALDI-TOF mass spectrometry analysis identified this proteasome co-isolated protein as ALAD. When the proteasome was isolated using density-gradient centrifugation, ALAD was also found in the 26S proteasome fractions. It co-isolated with the 20S more than with the 26S proteasome. Furthermore, immunoprecipitated ALAD stained positive with antibodies to proteasome subunits. These results indicate that ALAD might interact with the proteasome. It is possible that ALAD is involved in modulating proteasome activity. When purified proteasomes were incubated with ALAD it was found that ALAD changes proteasome activity in a dose dependent manner. This indicates that ALAD may play a significant role in regulating proteasome activity. The data supports the hypothesis that ALAD, an important enzyme for heme synthesis, is also important as a proteasome interacting protein. PMID- 21640721 TI - Epidermal growth factor increases LRF/Pokemon expression in human prostate cancer cells. AB - Leukemia/lymphoma related factor/POK erythroid myeloid ontogenic factor (LRF/Pokemon) is a member of the POK family of proteins that promotes oncogenesis in several forms of cancer. Recently, we found higher LRF expression in human breast and prostate carcinomas compared to the corresponding normal tissues. The aim of this study was to examine the regulation of LRF expression in human prostate cells. Epidermal growth factor (EGF) and its receptors mediate several tumorigenic cascades that regulate cell differentiation, proliferation, migration and survival of prostate cancer cells. There was significantly higher level of LRF expression in the nucleus of LNCaP and PC-3 cells than RWPE-1 cells. A significant increase in LRF expression was observed with increasing doses of EGF in more aggressive and androgen-sensitive prostate cancer cells suggesting that EGF signaling pathway is critical in upregulating the expression of LRF/Pokemon to promote oncogenesis. PMID- 21640722 TI - Xeroderma pigmentosum variant: complementary molecular approaches to detect a 13 base pair deletion in the DNA polymerase eta gene. AB - Deficiencies of DNA polymerase eta-an enzyme mediating replication past UV induced DNA damage-predispose individuals to xeroderma pigmentosum variant (XPV) and result in a high incidence of skin cancers. We designed, developed and assessed several complementary molecular approaches to detect a genetically inherited deletion within DNA polymerase eta. RNA was reverse transcribed from XPV fibroblasts and from normal human cells, and standard polymerase chain reaction (PCR) was conducted on the cDNA targeting a region with a 13 base pair deletion within the polymerase eta gene. PCR products were subjected to restriction fragment length polymorphism (RFLP) analysis and cycle DNA sequencing. The deletion was found to eliminate a BsrGI restriction site and affected the number of resultant fragments visualized after gel electrophoresis. Cycle sequencing of polymerase eta-specific amplicons from XPV and normal cells provided a second approach for detecting the mutation. Additionally, the use of a fluorescent nucleic acid dye-EvaGreen-in real-time PCR and melt curve analysis distinguished normal and XPV patient-derived amplicons as well as heteroduplexes that represent heterozygotic carriers without the need for high resolution melt analysis-compatible software. Our approaches are easily adaptable by diagnostic laboratories that screen for or verify genetically inherited disorders and identify carriers of a defective gene. PMID- 21640723 TI - Inhibition of adipocytogenesis by canonical WNT signaling in human mesenchymal stem cells. AB - The WNT signaling pathway plays important roles in the self-renewal and differentiation of mesenchymal stem cells (MSCs). Little is known about WNT signaling in adipocyte differentiation of human MSCs. In this study, we tested the hypothesis that canonical and non-canonical WNTs differentially regulate in vitro adipocytogenesis in human MSCs. The expression of adipocyte gene PPARgamma2, lipoprotein lipase, and adipsin increased during adipocytogenesis of hMSCs. Simultaneously, the expression of canonical WNT2, 10B, 13, and 14 decreased, whereas non-canonical WNT4 and 11 increased, and WNT5A was unchanged. A small molecule WNT mimetic, SB-216763, increased accumulation of beta-catenin protein, inhibited induction of WNT4 and 11 and inhibited adipocytogenesis. In contrast, knockdown of beta-catenin with siRNA resulted in spontaneous adipocytogenesis. These findings support the view that canonical WNT signaling inhibits and non-canonical WNT signaling promotes adipocytogenesis in adult human marrow-derived mesenchymal stem cells. PMID- 21640724 TI - Levodopa and Parkinson disease--electrophysiological perspectives in animal models. AB - Chronic administration of levodopa, while producing an "awakening" in patients with Parkinson disease (PD), causes disabling side effects such as motor fluctuations and dyskinesia. Indeed the most common reason for deep brain stimulation (DBS) in patients with PD is for treating these complications. However, our understanding of the complexities of these complications is still in the early days. Animal models (primate and rodent) have been exceedingly helpful in elucidating some of the mechanisms. More work needs to be done. In the paper by Gilmour et al. (2011) the authors have investigated the neuronal firing properties and local field potentials of two basal ganglia structures, in response to chronic treatment, to tackle this very question. This commentary attempts to place the work in the context of PD animal models, electrophysiology and where we need to go. PMID- 21640725 TI - p125/Sec23-interacting protein (Sec23ip) is required for spermiogenesis. AB - p125/Sec23ip is a phospholipase A(1)-like protein that interacts with Sec23, a coat component of COPII vesicles that bud from endoplasmic reticulum exit sites. To understand its physiological function, we produced p125 knockout mice. The p125 knockout mice grew normally, but males were subfertile. Sperm from p125 deficient mice had round heads and lacked the acrosome, an organelle containing the enzymes responsible for fertilization. p125 was found to be expressed at stages I-XII of spermatogenesis, similar to the expression pattern of proteins involved in acrosome biogenesis. These results suggest that p125 plays an important role in spermiogenesis. PMID- 21640726 TI - Water loss in insects: an environmental change perspective. AB - In the context of global environmental change much of the focus has been on changing temperatures. However, patterns of rainfall and water availability have also been changing and are expected to continue doing so. In consequence, understanding the responses of insects to water availability is important, especially because it has a pronounced influence on insect activity, distribution patterns, and species richness. Here we therefore provide a critical review of key questions that either are being or need to be addressed in this field. First, an overview of insect behavioural responses to changing humidity conditions and the mechanisms underlying sensing of humidity variation is provided. The primary sensors in insects belong to the temperature receptor protein superfamily of cation channels. Temperature-activated transient receptor potential ion channels, or thermoTRPs, respond to a diverse range of stimuli and may be a primary integrator of sensory information, such as environmental temperature and moisture. Next we touch briefly on the components of water loss, drawing attention to a new, universal model of the water costs of gas exchange and its implications for responses to a warming, and in places drying, world. We also provide an overview of new understanding of the role of the sub-elytral chamber for water conservation, and developments in understanding of the role of cuticular hydrocarbons in preventing water loss. Because of an increasing focus on the molecular basis of responses to dehydration stress we touch briefly on this area, drawing attention to the role of sugars, heat shock proteins, aquaporins, and LEA proteins. Next we consider phenotypic plasticity or acclimation responses in insect water balance after initial exposures to altered humidity, temperature or nutrition. Although beneficial acclimation has been demonstrated in several instances, this is not always the case. Laboratory studies show that responses to selection for enhanced ability to survive water stress do evolve and that genetic variation for traits underlying such responses does exist in many species. However, in others, especially tropical, typically narrowly distributed species, this appears not to be the case. Using the above information we then demonstrate that habitat alteration, climate change, biological invasions, pollution and overexploitation are likely to be having considerable effects on insect populations mediated through physiological responses (or the lack thereof) to water stress, and that these effects may often be non-intuitive. PMID- 21640727 TI - Expression of slow skeletal TnI in adult mouse hearts confers metabolic protection to ischemia. AB - Changes in metabolic and myofilament phenotypes coincide in developing hearts. Posttranslational modification of sarcomere proteins influences contractility, affecting the energetic cost of contraction. However, metabolic adaptations to sarcomeric phenotypes are not well understood, particularly during pathophysiological stress. This study explored metabolic adaptations to expression of the fetal, slow skeletal muscle troponin I (ssTnI). Hearts expressing ssTnI exhibited no significant ATP loss during 5 min of global ischemia, while non-transgenic littermates (NTG) showed continual ATP loss. At 7 min ischemia TG-ssTnI hearts retained 80+/-12% of ATP versus 49+/-6% in NTG (P<0.05). Hearts expressing ssTnI also had increased AMPK phosphorylation. The mechanism of ATP preservation was augmented glycolysis. Glycolytic end products (lactate and alanine) were 38% higher in TG-ssTnI than NTG at 2 min and 27% higher at 5 min. This additional glycolysis was supported exclusively by exogenous glucose, and not glycogen. Thus, expression of a fetal myofilament protein in adult mouse hearts induced elevated anaerobic ATP production during ischemia via metabolic adaptations consistent with the resistance to hypoxia of fetal hearts. The general findings hold important relevance to both our current understanding of the association between metabolic and contractile phenotypes and the potential for invoking cardioprotective mechanisms against ischemic stress. This article is part of a Special Issue entitled "Possible Editorial". PMID- 21640728 TI - Carbon monoxide: a vital signalling molecule and potent toxin in the myocardium. AB - Endogenous carbon monoxide (CO) is generated through the heme oxygenase-catalysed degradation of heme and is now established as an important, biologically active molecule capable of modulating a number of signalling pathways. Such pathways include those involving nitric oxide/guanylate cyclase, reactive oxygen species (ROS) and MAP kinases. In the heart, up-regulation of the inducible form of heme oxygenase (HO-1) following stresses such as ischemia/reperfusion provides cardioprotection, and much evidence indicates that CO accounts for many of these beneficial effects. One target of CO appears to be the L-type Ca(2+) channel; CO inhibits recombinant and native forms of this cardiac channel via mitochondria derived ROS, which likely contributes to the protective effects of CO. In stark contrast, exposure to exogenous CO is toxic: chronic, low-level exposure can lead to myocardial injury and fibrosis, whereas acute exposure is associated with life threatening arrhythmias. The molecular mechanisms accounting for such effects remain to be elucidated, but require future study before the potentially beneficial effects of CO therapy can be safely exploited. This article is part of a Special Issue entitled "Local Signaling in Myocytes". PMID- 21640729 TI - The effect of temporal variability on persistence conditions in rivers. AB - There has been great interest in the invasion and persistence of algal and insect populations in rivers. Recent modeling approaches assume that the flow speed of the river is constant. In reality, however, flow speeds in rivers change significantly on various temporal scales due to seasonality, weather conditions, or many human activities such as hydroelectric dams. In this paper, we study persistence conditions by deriving the upstream invasion speed in simple reaction advection-diffusion equations with coefficients chosen to be periodic step functions. The key methodological idea to determine the spreading speed is to use the exponential transform in order to obtain a moment generating function. In a temporally periodic environment, the averages of each coefficient function determine the minimal upstream and downstream propagation speeds for a single compartment model. For a two-compartment model, the temporal variation can enhance population persistence. PMID- 21640730 TI - The importance of cyclooxygenase 2-mediated oxidative stress in obesity-induced muscular insulin resistance in high-fat-fed rats. AB - AIM: This study was undertaken to examine the effect of cyclooxygenase (COX) 2 inhibition on the development of muscular insulin resistance in high-fat-induced obese rats. MAIN METHODS: The rats were on a regular chow diet (C) or high-fat enriched diet (HFD) energy-restrictedly (HFr), or ad libitum (HFa) for 12weeks. The rats fed HFD ad libitum were further divided into 3 groups: oral gavage with vehicle (HFa), selective COX-2 inhibitors-celecoxib (HFa+C) or nimesulid (HFa+N), 30mg/kg/day, respectively. KEY FINDINGS: Increased fasting plasma insulin, triglyceride and 8-isoprostane levels in HFa were significantly suppressed in those of HFa+C and HFa+N. The whole body insulin resistance of HFa indicated by the increased fasting plasma insulin levels and the elevated area under curve of insulin obtained from the oral glucose tolerance test were significantly reversed in those combined with celecoxib and nimesulid administration compared with those in HFr. The gene expression of COX-2 was significantly increased in epididymal fat but not in soleus muscle in HFa and the enhanced adipose COX-2 expression in high-fat fed rats was suppressed by those with drug treatment. Both selective COX 2 inhibitors reversed the diminished insulin-stimulated glucose uptake and GLUT4 translocation in skeletal muscles of HFa. Obesity-induced oxidative stress indicated by the elevated plasma 8-isoprostane,the decreased ratio of GSH/GSSG and increased TBARS in soleus muscle were significantly reversed by COX-2 inhibition. SIGNIFICANCE: The results suggest that COX-2 inhibition might suppress the muscular insulin resistance indirectly through decreasing the COX-2 mediated systemic oxidative stress in this diet-induced obese model. PMID- 21640731 TI - Predicting dynamics and rheology of blood flow: A comparative study of multiscale and low-dimensional models of red blood cells. AB - We compare the predictive capability of two mathematical models for red blood cells (RBCs) focusing on blood flow in capillaries and arterioles. Both RBC models as well as their corresponding blood flows are based on the dissipative particle dynamics (DPD) method, a coarse-grained molecular dynamics approach. The first model employs a multiscale description of the RBC (MS-RBC), with its membrane represented by hundreds or even thousands of DPD-particles connected by springs into a triangular network in combination with out-of-plane elastic bending resistance. Extra dissipation within the network accounts for membrane viscosity, while the characteristic biconcave RBC shape is achieved by imposition of constraints for constant membrane area and constant cell volume. The second model is based on a low-dimensional description (LD-RBC) constructed as a closed torus-like ring of only 10 large DPD colloidal particles. They are connected into a ring by worm-like chain (WLC) springs combined with bending resistance. The LD RBC model can be fitted to represent the entire range of nonlinear elastic deformations as measured by optical-tweezers for healthy and for infected RBCs in malaria. MS-RBCs suspensions model the dynamics and rheology of blood flow accurately for any vessel size but this approach is computationally expensive for vessel diameters above 100MUm. Surprisingly, the much more economical suspensions of LD-RBCs also capture the blood flow dynamics and rheology accurately except for small-size vessels comparable to RBC diameter. In particular, the LD-RBC suspensions are shown to properly capture the experimental data for the apparent viscosity of blood and its cell-free layer (CFL) in tube flow. Taken together, these findings suggest a hierarchical approach in modeling blood flow in the arterial tree, whereby the MS-RBC model should be employed for capillaries and arterioles below 100MUm, the LD-RBC model for arterioles, and the continuum description for arteries. PMID- 21640732 TI - The effect of memantine in harmaline-induced tremor and neurodegeneration. AB - Essential tremor (ET) is one of the most common and most disabling movement disorders among adults. The drug treatment of ET remains unsatisfactory. Additional therapies are required for patients with inadequate response or intolerable side effects. The current study aims to investigate the anti tremogenic and neuroprotective effects of memantine (NMDA receptor antagonist) on the harmaline model of transient action tremor. The effects of memantine were further compared with ethanol. Three separate groups of male Wistar rats were injected either with saline, ethanol (1.5 gr/kg), or memantine (5 mg/kg) 15 min prior to a single intraperitoneal injection of harmaline (20 mg/kg). Tremor and locomotion were evaluated by a custom-built tremor and locomotion analysis system. After 24 h of harmaline injection, cellular viability, and apoptosis were assessed using crystal violet staining, and caspase-3 immunostaining, respectively. Harmaline caused neuronal cell loss and caspase-3 mediated apoptosis in cerebellar granular and purkinje cells as well as the inferior olivary neurons. Despite a reduction in tremor intensity and duration with ethanol, this compound resulted in cell loss in cerebellum and olivary nucleus. Memantine exhibited neuroprotective efficacy on cerebellar and inferior olivary neurons albeit weaker anti-tremor effect compared to ethanol. In conclusion, anti tremogenic and neuroprotective effects do not necessarily overlap. Memantine is a potential treatment for ET particularly given its neuroprotective efficacy. PMID- 21640733 TI - The wake-promoting transmitter histamine preferentially enhances alpha-4 subunit containing GABAA receptors. AB - Histamine is an important wake-promoting neurotransmitter that activates seven transmembrane G-protein coupled histamine receptors. However, histamine demonstrates target promiscuity, including direct interaction with the structurally unrelated glutamate (NMDA) and GABA(A) receptor channels. Previous work showed that histamine enhances the activity of recombinant GABA(A) receptor isoforms typically found in synaptic locations, although co-release of histamine and GABA is not known to occur in vivo. Here we used patch clamp recordings of various recombinant GABA(A) receptor isoforms (alpha1-6, beta1-3, gamma1-3, delta) to test the hypothesis that histamine might show subunit preference under low GABA concentration (extrasynaptic) conditions. We found that histamine potentiated the whole-cell responses to GABA for all tested subunit combinations. However, the magnitude of enhancement was largest (~400% of EC(10) GABA-evoked currents) with alpha4beta3 and alpha4beta3X isoforms, where X could be gamma or delta. In contrast, histamine (1 mM) had small effects on prolonging deactivation of alpha4beta3gamma2 receptors following brief (5 ms) pulses of 1 mM GABA. These findings suggest GABA-histamine cross-talk may occur preferentially at low GABA concentrations, which could theoretically be inhibitory (via enhancing tonic inhibition), directly excitatory (via enhancing presynaptic GABAergic signaling), or indirectly excitatory (via inhibiting GABAergic interneurons). PMID- 21640734 TI - Differential effect of the mGlu5 receptor positive allosteric modulator ADX-47273 on early and late hippocampal LTP. AB - Conflicting findings are reported in the literature about the involvement of the mGlu5 receptor in hippocampal long-term potentiation (LTP), which might be a consequence of different sub-types of LTP induced by the investigators due to the specific experimental conditions used. A comparable controversy came up in the past concerning the influence of different experimental conditions on the involvement of L-type voltage dependent calcium channels (L-VDCCs) and NMDA receptors in hippocampal LTP. In this study, two stimulation protocols with otherwise identical conditions were used to probe modulatory effects of mGlu5 receptor activation in NMDA receptor and L-VDCCs dependent CA1 LTP: weak high frequency stimulation (20 stimuli at 100 Hz) to induce early LTP and repeated strong high frequency stimulation (3 times 100 stimuli at 100 Hz with 5 min interval) to induce late LTP, which - in contrast to early LTP - was shown to be protein-synthesis dependent. Using the NMDA receptor antagonist MK-801 and the L type calcium channel blocker nifedipine, early LTP was shown to be dependent on NMDA receptors only, whereas late LTP was demonstrated to be dependent on NMDA receptors and L-VDCCs in about equal parts. Moreover, late LTP, but not early LTP, was increased by the mGlu5 receptor positive allosteric modulator ADX-47273, indicating that artificial augmentation of mGlu5 receptor activation by endogenous glutamate may boost the protein-synthesis dependent form of LTP but not the protein-synthesis independent form. PMID- 21640735 TI - Sex difference in sensitivity to allopregnanolone neuroprotection in mice correlates with effect on spontaneous inhibitory post synaptic currents. AB - Allopregnanolone (ALLO) is a neurosteroid that has many functions in the brain, most notably neuroprotection and modulation of gamma-amino butyric acid (GABA) neurotransmission. Using a mouse model of cardiac arrest and cardiopulmonary resuscitation, we have previously demonstrated that ALLO protects cerebellar Purkinje cells (PCs) from ischemia in a GABA(A) receptor-dependent manner. In this study we examined the effect of sex on ALLO neuroprotection, observing that low dose ALLO (2 mg/kg) provided greater neuroprotection in females compared to males. At a higher dose of ALLO (8 mg/kg), both sexes were significantly protected from ischemic damage. Using an acute cerebellar slice preparation, whole cell voltage clamp recordings were made from PCs. Spontaneous inhibitory post synaptic currents (IPSCs) were analyzed and the response to physiological ALLO (10 nM) was significantly greater in female PCs compared to male. In contrast, recordings of miniature IPSCs, did not exhibit a sex difference in response to ALLO, suggesting that ALLO affects males and females differentially through a mechanism other than binding postsynaptic GABA(A) receptors. We conclude that the female brain has greater sensitivity to ALLO mediated potentiation of GABAergic neurotransmission, contributing to increased neuroprotection. PMID- 21640736 TI - Chronic phencyclidine (PCP)-induced modulation of muscarinic receptor mRNAs in rat brain: impact of antipsychotic drug treatment. AB - Many antipsychotics (APDs) have a high affinity for muscarinic receptors, which is thought to contribute to their therapeutic efficacy, or side effect profile. In order to define how muscarinic receptor gene expression is affected by atypical or typical APDs, rats were treated with chronic (2.58 mg/kg) PCP (a psychotomimetic) or vehicle, plus clozapine (20 mg/kg/day) or haloperidol (1 mg/kg/day), and M1, M2 and M3 receptor mRNA levels were determined in brain sections. Negligible changes in M2 or M3 muscarinic mRNA were detected in any region after clozapine or haloperidol. Chronic PCP administration increased M1 mRNA expression in the prefrontal cortex, which was not reversed by either chronic clozapine or haloperidol treatment. Chronic clozapine treatment in combination with PCP treatment decreased M1 receptor mRNA levels in the nucleus accumbens core, whereas chronic haloperidol in combination with PCP treatment increased M1 receptor mRNA levels in the ventromedial hypothalamus and medial amygdala. Thus M1 receptor gene expression is targeted by APDs, although the regions affected differ according to the APD treatment and whether PCP has been administered. The different brain circuitry modulated, may reflect the differing modes of action of typical and atypical APDs. These data provide support for the dysregulation of M1 receptors in schizophrenia, and furthermore, modulation by antipsychotic agents in the treatment of schizophrenia. PMID- 21640737 TI - ERP evidence for selective drop in attentional costs in uncertain environments: challenging a purely premotor account of covert orienting of attention. AB - Several studies have proved that the reliability of endogenous spatial cues linearly modulates the reaction time advantage in the processing of targets at validly cued vs. invalidly cued locations, i.e. the "validity effect". This would imply that with non-predictive cues, no "validity effect" should be observed. However, contrary to this prediction, one could hypothesize that attentional benefits by valid cuing (i.e. the RT advantage for validly vs. neutrally cued targets) can still be maintained with non-predictive cues, if the brain were endowed with mechanisms allowing the selective reduction in costs of reorienting from invalidly cued locations (i.e. the reduction of the RT disadvantage for invalidly vs. neutrally cued targets). This separated modulation of attentional benefits and costs would be adaptive in uncertain contexts where cues predict at chance level the location of targets. Through the joint recording of manual reaction times and event-related cerebral potentials (ERPs), we have found that this is the case and that relying on non-predictive endogenous cues results in abatement of attentional costs and the difference in the amplitude of the P1 brain responses evoked by invalidly vs. neutrally cued targets. In contrast, the use of non-predictive cues leaves unaffected attentional benefits and the difference in the amplitude of the N1 responses evoked by validly vs. neutrally cued targets. At the individual level, the drop in costs with non-predictive cues was matched with equivalent lateral biases in RTs to neutrally and invalidly cued targets presented in the left and right visual field. During the cue period, the drop in costs with non-predictive cues was preceded by reduction of the Early Directing Attention Negativity (EDAN) on posterior occipital sites and by enhancement of the frontal Anterior Directing Attention Negativity (ADAN) correlated to preparatory voluntary orienting. These findings demonstrate, for the first time, that the segregation of mechanisms regulating attentional benefits and costs helps efficiency of orienting in "uncertain" visual spatial contexts characterized by poor probabilistic association between cues and targets. PMID- 21640738 TI - Interhemispheric integration in visual search. AB - The search task of Luck, Hillyard, Mangun and Gazzaniga (1989) was optimised to test for the presence of a bilateral field advantage in the visual search capabilities of normal subjects. The modified design used geometrically regular arrays of 2, 4 or 8 items restricted to hemifields delineated by the vertical or horizontal meridian; the target, if present, appeared at one of two fixed positions per quadrant at an eccentricity of 11 deg. Group and individual performance data were analysed in terms of the slope of response time against display-size functions ('RT slope'). Averaging performance across all conditions save display mode (bilateral vs. unilateral) revealed a significant bilateral advantage in the form of a 21% increase in apparent item scanning speed for target detection; in the absence of a target, bilateral displays gave a 5% increase in speed that was not significant. Factor analysis by ANOVA confirmed this main effect of display mode, and also revealed several higher order interactions with display geometry, indicating that the bilateral advantage was masked at certain target positions by a crowding-like effect. In a numerical model of search efficiency (i.e. RT slope), bilateral advantage was parameterised by an interhemispheric 'transfer factor' (T) that governs the strength of the ipsilateral representation of distractors, and modifies the level of intrahemispheric competition with the target. The factor T was found to be higher in superior field than inferior field; this result held for the modelled data of each individual subject, as well as the group, representing a uniform tendency for the bilateral advantage to be more prominent in inferior field. In fact statistical analysis and modelling of search efficiency showed that the geometrical display factors (target polar and quadrantic location, and associated crowding effects) were all remarkably consistent across subjects. Greater variability was inferred within a fixed, decisional component of response time, with individual subjects capable of opposite hemifield biases. The results are interpretable by a guided search model of spatial attention - a first, parallel stage guiding selection by a second, serial stage - with the proviso that the first stage is relatively insular within each hemisphere. The bilateral advantage in search efficiency can then be attributed to a relative gain in target weight within the initial parallel stage, owing to a reduction in distractor competition mediated specifically by intrahemispheric circuitry. In the absence of a target there is no effective guidance, and hence no basis for a bilateral advantage to enhance search efficiency; the equivalence of scanning speed for the two display modes (bilateral and unilateral) implies a unitary second-stage process mediated via efficient interhemispheric integration. PMID- 21640739 TI - Anomalies in social behaviors and exploratory activities in an APPswe/PS1 mouse model of Alzheimer's disease. AB - Alzheimer's disease is characterized by deficits in social communication, associated with generalized apathy or agitation, as well as social memory. To assess social behaviors in 6-month-old male APPswe/PS1 bigenics relative to non transgenic controls, the 3-chamber test was used, together with open-field and elevated plus-maze tests of exploration. APPswe/PS1 mice were less willing to engage in social interaction than wild-type, avoiding an unfamiliar stimulus mouse, probably not due to generalized apathy because in both tests of exploratory activity the mutants were hyperactive. This study reveals reduced "sociability" combined with hyperactivity in an APPswe/PS1 mouse model of Alzheimer dementia. PMID- 21640740 TI - Effects of the potassium ion channel modulators BMS-204352 Maxipost and its R enantiomer on salicylate-induced tinnitus in rats. AB - Currently, there are no effective pharmacological therapies for chronic tinnitus despite a number of efforts from clinical studies and more recently, studies in animals using compounds to enhance endogenous inhibition or reduce central hyperactivity. The purpose of the current study was to evaluate the therapeutic efficacy of a novel anxiolytic with potassium channel activity in suppressing salicylate induced tinnitus in animals. Kv7 potassium channels are present in the peripheral and central auditory system where they are believed to modulate neural activity. Maxipost, a compound which attenuates hyperexcitability via positive modulation of Kv7.2-Kv7.5 channels, was administered to rats with behavioral evidence of salicylate induced tinnitus. Tinnitus was measured using our previously established animal model, Schedule Induced Polydipsia Avoidance Conditioning, a paradigm where rats were conditioned to drink only during quiet and suppress drinking in the presence of sound. Salicylate alone significantly suppressed licks in quiet but had no effect on licks in sound; results consistent with the presence of tinnitus. Maxipost at 10 mg/kg suppressed behavioral evidence of tinnitus as it completely reversed salicylate's suppression of licks in quiet. Unexpectedly, the R-enantiomer of Maxipost, R-Maxipost, which has no anxiolytic effects and negatively modulates Kv7.2-Kv7.5, also suppressed behavioral evidence of tinnitus. Our original hypothesis was that Kv7.2-Kv7.5 channels might play a key role in tinnitus generation and that Maxipost but not R Maxipost would suppress tinnitus; however, it appears that a shared mechanism between Maxipost and R-xMaxipost, such as inhibition of Kv7.1 channels or activation of BK channels or some novel mechanism common to both compounds, underlies salicylate induced tinnitus as both compounds completely abolished behavioral evidence of tinnitus in a dose-dependent manner. Further studies with specific BK channel agonists/antagonists are necessary to determine the contribution of these channels to other forms of tinnitus or determine novel targets that could be related to tinnitus. PMID- 21640741 TI - Synthesis and acetylcholinesterase inhibitory activity of 2beta,3alpha-disulfoxy 5alpha-cholestan-6-one. AB - Disodium 2beta,3alpha-dihydroxy-5alpha-cholestan-6-one disulfate (8) has been synthesized using cholesterol (1) as starting material. Sulfation was performed using trimethylamine-sulfur trioxide complex in dimethylformamide as the sulfating agent. The acetylcholinesterase inhibitory activity of compound 8 was evaluated and compared to that of disodium 2beta,3alpha-dihydroxy-5alpha cholestane disulfate (10) and diols 7 and 9. Compounds 8 and 10 were active with IC(50) values of 14.59 and 59.65 MUM, respectively. Diols 7 and 9 showed no inhibitory activity (IC(50)>500 MUM). PMID- 21640742 TI - Mathematical properties of Fst between admixed populations and their parental source populations. AB - We consider the properties of the F(st) measure of genetic divergence between an admixed population and its parental source populations. Among all possible populations admixed among an arbitrary set of parental populations, we show that the value of F(st) between an admixed population and a specific source population is maximized when the admixed population is simply the most distant of the other source populations. For the case with only two parental populations, as a function of the admixture fraction, we further demonstrate that this F(st) value is monotonic and convex, so that F(st) is informative about the admixture fraction. We illustrate our results using example human population-genetic data, showing how they provide a framework in which to interpret the features of F(st) in admixed populations. PMID- 21640743 TI - Metabolomics analysis reveals elevation of 3-indoxyl sulfate in plasma and brain during chemically-induced acute kidney injury in mice: investigation of nicotinic acid receptor agonists. AB - An investigative renal toxicity study using metabolomics was conducted with a potent nicotinic acid receptor (NAR) agonist, SCH 900424. Liquid chromatography mass spectrometry (LC-MS) and gas chromatography-mass spectrometry (GC-MS) techniques were used to identify small molecule biomarkers of acute kidney injury (AKI) that could aid in a better mechanistic understanding of SCH 900424-induced AKI in mice. The metabolomics study revealed 3-indoxyl sulfate (3IS) as a more sensitive marker of SCH 900424-induced renal toxicity than creatinine or urea. An LC-MS assay for quantitative determination of 3IS in mouse matrices was also developed. Following treatment with SCH 900424, 3IS levels were markedly increased in murine plasma and brain, thereby potentially contributing to renal- and central nervous system (CNS)-related rapid onset of toxicities. Furthermore, significant decrease in urinary excretion of 3IS in those animals due to compromised renal function may be associated with the elevation of 3IS in plasma and brain. These data suggest that 3IS has a potential to be a marker of renal and CNS toxicities during chemically-induced AKI in mice. In addition, based on the metabolomic analysis other statistically significant plasma markers including p-cresol-sulfate and tryptophan catabolites (kynurenate, kynurenine, 3-indole lactate) might be of toxicological importance but have not been studied in detail. This comprehensive approach that includes untargeted metabolomic and targeted bioanalytical sample analyses could be used to investigate toxicity of other compounds that pose preclinical or clinical development challenges in a pharmaceutical discovery and development. PMID- 21640744 TI - AKR1B10 induces cell resistance to daunorubicin and idarubicin by reducing C13 ketonic group. AB - Daunorubicin, idarubicin, doxorubicin and epirubicin are anthracyclines widely used for the treatment of lymphoma, leukemia, and breast, lung, and liver cancers, but tumor resistance limits their clinical success. Aldo-keto reductase family 1 B10 (AKR1B10) is an NADPH-dependent enzyme overexpressed in liver and lung carcinomas. This study was aimed to determine the role of AKR1B10 in tumor resistance to anthracyclines. AKR1B10 activity toward anthracyclines was measured using recombinant protein. Cell resistance to anthracycline was determined by ectopic expression of AKR1B10 or inhibition by epalrestat. Results showed that AKR1B10 reduces C13-ketonic group on side chain of daunorubicin and idarubicin to hydroxyl forms. In vitro, AKR1B10 converted daunorubicin to daunorubicinol at V(max) of 837.42+/-81.39nmol/mg/min, K(m) of 9.317+/-2.25mM and k(cat)/K(m) of 3.24. AKR1B10 showed better catalytic efficiency toward idarubicin with V(max) at 460.23+/-28.12nmol/mg/min, K(m) at 0.461+/-0.09mM and k(cat)/K(m) at 35.94. AKR1B10 was less active toward doxorubicin and epirubicin with a C14-hydroxyl group. In living cells, AKR1B10 efficiently catalyzed reduction of daunorubicin (50nM) and idarubicin (30nM) to corresponding alcohols. Within 24h, approximately 20+/-2.7% of daunorubicin (1MUM) or 23+/-2.3% of idarubicin (1MUM) was converted to daunorubicinol or idarubicinol in AKR1B10 expression cells compared to 7+/ 0.9% and 5+/-1.5% in vector control. AKR1B10 expression led to cell resistance to daunorubicin and idarubicin, but inhibitor epalrestat showed a synergistic role with these agents. Together our data suggest that AKR1B10 participates in cellular metabolism of daunorubicin and idarubicin, resulting in drug resistance. These data are informative for the clinical use of idarubicin and daunorubicin. PMID- 21640746 TI - Pascal's ring, cardinal points, and refractive compensation. AB - Pascal's ring is a hexagon each of whose corners represents one of the six cardinal points of an optical system and whose sides represent relationships of relative axial position of the cardinal points. Changes to the ring represent the axial displacements of the cardinal points of the visual optical system of an eye that are caused when a spectacle lens compensates for the ametropia. Pascal's schema was described some 70 years ago with little theoretical justification. The purpose of this paper is to derive expressions for the axial locations of the cardinal points of a compound system consisting of an optical instrument and a visual optical system and for the shift caused by the instrument, and to provide theoretical justification for Pascal's schema. The cardinal points are treated not as separate entities but in a unified manner as special cases of an infinite class of special points. Expressions are derived using Gaussian optics. The results are specialized for the case of the eye's ametropia compensated by optical instruments in general and by spectacle lenses in particular. Pascal's schema is shown to be broadly correct although some modification is necessary for the effects on the incident cardinal points especially for the myopic eye. PMID- 21640745 TI - Phylogenetic analysis of serine proteases from Russell's viper (Daboia russelli siamensis) and Agkistrodon piscivorus leucostoma venom. AB - Serine proteases are widely found in snake venoms. They have variety of functions including contributions to hemostasis. In this study, five serine proteases were cloned and characterized from two different cDNA libraries: factor V activator (RVV-V), alpha fibrinogenase (RVAF) and beta fibrinogenase (RVBF) from Russell's viper (Daboia russelli siamensis), and plasminogen activator (APL-PA) and protein C activator (APL-C) from Agkistrodon piscivorus leucostoma. The snake venom serine proteases were clustered in phylogenetic tree according to their functions. K(A)/K(S) values suggested that accelerated evolution has occurred in the mature protein coding regions in cDNAs of snake venom serine proteases. PMID- 21640747 TI - Discrimination of natural scenes in central and peripheral vision. AB - We conducted suprathreshold discrimination experiments to compare how natural scene information is processed in central and peripheral vision (16 degrees eccentricity). Observers' ratings of the perceived magnitude of changes in naturalistic scenes were lower for peripheral than for foveal viewing, and peripheral orientation changes were rated less than peripheral colour changes. A V1-based Visual Difference Predictor model of the magnitudes of perceived foveal change was adapted to match the sinusoidal grating sensitivities of peripheral vision, but it could not explain why the ratings for changes in peripheral stimuli were so reduced. Perceived magnitude ratings for peripheral stimuli were further reduced by simultaneous presentation of flanking patches of naturalistic images, a phenomenon that could not be replicated foveally, even after M-scaling the foveal stimuli to reduce their size and the distances from the flankers. The effects of the peripheral flankers are very reminiscent of crowding phenomena demonstrated with letters or Gabor patches. PMID- 21640748 TI - Subeffective doses of nitroparacetamol (NCX-701) enhance the antinociceptive activity of the alpha2-adrenoceptor agonist medetomidine. AB - The alpha2-adrenergic system is involved in pain processing and inflammation induced sensitization. alpha2-adrenoceptor agonists induce analgesia, and this effect is greater when administered in combination with other analgesics. In the present study, we assessed a possible enhancement of antinociception combining the alpha2-adrenoceptor agonist medetomidine with subeffective doses of NCX701 (nitroparacetamol). The effects of the drugs were studied in spinal cord neuronal responses from adult male Wistar rats with carrageenan-induced inflammation, using the recording of single motor unit technique. The experiments showed that the i.v. administration of medetomidine and NCX701 induced a more potent and effective antinociceptive effect than medetomidine when given alone (ID50: 0.47+/ 0.1 vs. 1.1+/-0.1 MUg/kg) or in the presence of paracetamol, in naturally-evoked nociceptive responses. In addition, the duration of antinociception was significantly longer (P<0.001, 100 min after administration). The use of low doses of NCX701 and alpha2-adrenoceptor agonists might open new perspectives in the treatment of inflammatory pain. PMID- 21640749 TI - Effects of pregnanolone and flunitrazepam on the retention of response sequences in rats. AB - Neuroactive steroids produce effects similar to other GABA(A) modulators (e.g., benzodiazepines and barbiturates) and have a large therapeutic potential; however, a greater understanding of the effects of these substances on learning and memory is needed. To specifically assess the effects of a neurosteroid on memory, pregnanolone (1-18 mg/kg) was administered to male Long-Evans rats responding under a repeated acquisition and delayed-performance procedure in which different 4-response sequences were acquired and then retested after varying delays. Responding was maintained under a second-order fixed-ratio (FR) 2 schedule of food reinforcement, and incorrect responses (errors) produced a 5-sec timeout. For comparison purposes, both a high (flunitrazepam) and low efficacy agonist/antagonist (flumazenil) of the GABA(A) receptor complex were also administered both alone and in combination. Retention of each sequence was quantified as percent savings in errors-to-criterion and this dependent measure was shown to be sensitive to increases in delay. When administered 15 min prior to the end of either a 30- or 180-minute delay, pregnanolone produced both dose- and delay-dependent decreases in percent savings, response rate and accuracy; this effect was selective in that decreases in retention occurred at doses lower than those that disrupted response rate or accuracy. Flunitrazepam (0.056-1mg/kg) produced similar disruptions in retention and these disruptions were antagonized by 5.6 mg/kg of flumazenil. Both an ineffective (0.056 mg/kg) and an effective (0.18 mg/kg) dose of flunitrazepam also potentiated the dose- and delay-dependent disruptions in retention produced by pregnanolone. These data indicate that the neurosteroid pregnanolone disrupts retention in a manner similar to the benzodiazepine flunitrazepam, and suggests that the interaction of flunitrazepam and pregnanolone on retention may be mediated by the GABA(A) receptor complex. PMID- 21640750 TI - Oral tremor induced by galantamine in rats: a model of the parkinsonian side effects of cholinomimetics used to treat Alzheimer's disease. AB - Anticholinesterases are the most common treatment for Alzheimer's disease, and, in recent years, a new group of cholinesterase inhibitors (i.e. rivastigmine, galantamine, and donepezil) has become available. Although these drugs improve cognitive symptoms, they also can induce or exacerbate parkinsonian symptoms, including tremor. The present studies were conducted to determine if galantamine induces tremulous jaw movements, a rodent model of parkinsonian tremor, and to investigate whether these oral motor impairments can be reversed by co administration of adenosine A(2A) antagonists. The first experiment demonstrated that systemic injections of galantamine (0.75-6.0 mg/kg I.P.) induced a dose related increase in tremulous jaw movements in rats. In a second study, co administration of the muscarinic antagonist scopolamine (0.0156-0.25 mg/kg I.P.) produced a dose dependent suppression of tremulous jaw movements induced by a 3.0 mg/kg dose of galantamine, indicating that galantamine induces these tremulous oral movements through actions on muscarinic acetylcholine receptors. In two additional studies, analyses of freeze-frame video and electromyographic activity recorded from the lateral temporalis muscle indicated that the local frequency of these galantamine-induced jaw movements occurs in the 3-7 Hz frequency range that is characteristic of parkinsonian tremor. In the final experiment, the adenosine A(2A) antagonist MSX-3 significantly attenuated the tremulous jaw movements induced by the 3.0mg/kg dose of galantamine, which is consistent with the hypothesis that co-administration of adenosine A(2A) antagonists may be beneficial in reducing parkinsonian motor impairments induced by anticholinesterase treatment. PMID- 21640751 TI - Size, structure and scaling relationships in glycogen from various sources investigated with asymmetrical flow field-flow fractionation and 1H NMR. AB - In this paper we investigate the size, structure and scaling relationships in glycogen isolated from five different animal sources. For this purpose a versatile fractionation technique, asymmetrical flow field-flow fractionation (AsFlFFF), coupled to multi-angle light scattering, is utilized. For determination of the average degree of branching (1)H NMR is utilized. The results give a detailed insight into the physico-chemical properties of glycogen over the whole size distribution. The results show that glycogen is a hyper branched macromolecule with wide size distributions, and in some samples two major populations are clearly observed which most likely correspond to beta- and alpha-particles of glycogen. The results also illustrates that glycogen is a polysaccharide showing rather diverse conformational properties, over the size distribution, depending on its origin and the extraction procedure. The ratio between root-mean-square radius and hydrodynamic radius varies depending of both sample origin the molar mass of the macromolecules, reflecting differences in conformation and scaling within the size distribution. Thus, a priori assumptions regarding the r(rms)/r(h) are difficult to make and r(rms)/r(h) based on average properties give an incomplete description of the properties. Furthermore, the results display the strength of the apparent density (as obtained from AsFlFFF MALS-RI) as a characterization parameter for scaling in disperse macromolecules. PMID- 21640752 TI - Characterization of edible emulsified films with low affinity to water based on kefiran and oleic acid. AB - New edible composite films based on kefiran and oleic acid (OA) at the ratio of 15, 25, and 35% (w/w) were prepared using emulsification with the aim of improving their water vapour barrier and mechanical properties. Film-forming solutions were characterized in terms of rheological properties and particle-size distribution. The impact of the incorporation of OA into the film matrix was studied by investigating the physical, mechanical, and thermal properties of the films. The water vapour permeability (WVP) of the emulsified films was reduced by approximately 33% by adding OA. The mechanical properties of kefiran films were also affected by adding OA: tensile strength was diminished, and elongation increased considerably. Differential scanning calorimetry showed that the glass transition temperature (T(g)) of the kefiran film was -16 degrees C and was not considerably affected by adding OA. Therefore, OA could be incorporated into these films for some food-technology applications that need a low affinity toward water. PMID- 21640753 TI - Rheological and microstructural investigation of oat beta-glucan isolates varying in molecular weight. AB - The rheological properties and microstructure of aqueous oat beta-glucan solutions varying in molecular weight were investigated. The structural features and molecular weights (MW) were characterized by (13)C NMR spectroscopy and high performance size-exclusion chromatography (HPSEC), respectively. The microstructure of the beta-glucans dispersions was also examined by atomic force microscopy (AFM). The samples with beta-glucan content between 78 and 86% on a dry weight basis had MW, intrinsic viscosity ([eta]) and critical concentration (c*) in the range of 142-2800*10(3)g/mol, 1.7-7.2dl/g and 0.25-1.10g/dl, respectively. The flow and viscoelastic behaviour was highly dependent on MW and on the concentration of the beta-glucans dispersions. Pseudoplastic behaviour was exhibited at high concentrations and Newtonian behaviour was evident at low concentrations. At the same concentration, the viscosity was higher for higher MW samples. The Cox-Merz rule was applicable for the lower molecular weight samples at higher concentrations whereas the high molecular weight sample deviated at concentrations greater than 1.0%, w/v. The mechanical spectra with variation of both MW and concentration were typical of entangled biopolymer solutions. AFM images revealed the formation of clusters or aggregates linked via individual polymer chains scattered heterogeneously throughout the system. The aggregate size increased with the molecular weight of the samples investigated and has been linked to the rheological behaviour of the samples. PMID- 21640754 TI - Isolation and antioxidant activities of polysaccharides extracted from the shoots of Phyllostachys edulis (Carr.). AB - Polysaccharides extracted from Phyllostachys edulis (Carr.) are a group of hetero polysaccharides, and their antioxidant activities were investigated employing various established in vitro systems. Available data obtained with in vitro models suggested that among the three samples, B1 (extraction with water) showed significant inhibitory effects on superoxide radical and hydroxyl radical; its reducing power was also the strongest among the three samples. These results clearly establish the possibility that polysaccharides extracted from P. edulis could be effectively employed as ingredient in health or functional food, to alleviate oxidative stress. However, comprehensive studies need to be conducted in experimental animal models. PMID- 21640755 TI - Antistress properties of antidepressant drugs and their clinical implications. AB - Stress, in its different forms, represents a major environmental component for increased susceptibility to mental illness and, indeed, stressful life events serve as significant predictors of depression. With this respect, since antidepressant drugs may be effective in a variety of conditions that are triggered by, or associated with, exposure to stress, it is important to establish to what extent drug treatment can interfere with behavioral, functional and molecular alterations induced by stress exposure. The aim of this article is to examine the evidence supporting the ability of antidepressant treatment to reverse or modulate stress-induced alterations in humans and animals. Biologically, there is accumulating evidence that antidepressants can modify not only the neuroendocrine response to stress, but also mechanisms that regulate neuronal plasticity and that may play a key role in structural and functional changes associated with major depression. We believe that a detailed analysis of pharmacotherapy effects on stress-induced alterations may be important to define the ability and potency of different antidepressants to 'normalize' alterations that are central to the disease, in order to differentiate them with respect to the pathologic phenotype. This may lead to the identification of new pharmacological targets and contribute to the development of novel and more effective agents, which may eventually achieve higher rates of remission. PMID- 21640756 TI - Current and future therapeutic strategies for functional repair of spinal cord injury. AB - Spinal cord injury (SCI) causes serious, chronic dysfunction which is difficult to treat. Disability, including long-lasting motor and sensory dysfunction, typically results from damage to the descending and ascending spinal tracts and interneurons and, secondarily, to the neuronal degeneration that occurs proximal and distal to the spinal insult. Numerous strategies are being implemented to protect neurons from damage, to enhance axon growth and to foster cell proliferation. Described in this report are recent clinical trials aimed at testing strategies to restore locomotion after SCI. While laboratory animal studies have indicated that it may be possible to minimize neuronal damage resulting from spinal cord injury, little progress has been made in reducing or reversing the events associated with the chronic phase of this condition. The strategy aiming to inhibit single molecule sometimes shows controversial results. In SCI, a lot of players participate in motor and sensory dysfunctions. Therefore, sufficient functional recovery may be achieved by regulating multiple targets. Regrowth of tracts connecting the brain and spinal cord, and axonal sprouting of propriospinal interneurons are fundamentally important for neuronal network working. In addition, remyelination, protection of neuronal death, inhibition of inflammation, and upregulation of beneficial influence of astrocytes are also quite crucial to supporting the axonal refining. Combination of several strategies might be useful as a practical therapy. Several compounds such as a Sema3A inhibitor, estrogen, withanoside IV and their relating compounds or other neurotrophic factor-mimicking agents may be candidates for useful SCI therapeutic drugs since those have multi-effects on damaged spinal cord. [corrected]. PMID- 21640757 TI - Vitamin D and metabolic health with special reference to the effect of vitamin D on serum lipids. AB - Considering that the vitamin D receptor as well as the 1-alpha-hydroxylase enzyme that converts 25-hydroxyvitamin D (25(OH)D) to its active form 1,25 dihydroxyvitamin D have been found in tissues throughout the body, it is likely that vitamin D is important for more than the calcium balance. Accordingly, low serum levels of 25(OH)D have been associated with mortality, cardiovascular disease, type 2 diabetes, hypertension and obesity. Low serum levels of 25(OH)D have also been associated with an unfavourable lipid profile, which could possible explain the relation with cardiovascular disease and mortality. However, the relation between vitamin D and lipids have so far received little attention and is therefore the main focus of the present review. A PubMed search identified 22 cross-sectional studies where serum levels of 25(OH)D and lipids were related and that included a minimum of 500 subjects, and 10 placebo-controlled double blind intervention studies with vitamin D where more than 50 subjects were included. In all the cross-sectional studies serum 25(OH)D was positively associated with high-density lipoprotein cholesterol (HDL-C) resulting in a favourable low-density lipoprotein cholesterol (LDL-C) (or total cholesterol) to HDL-C ratio. There was also a uniform agreement between studies on a negative relation between serum 25(OH)D and triglycerides (TG). On the other hand, the intervention studies gave divergent results, with some showing a positive and some a negative effect of vitamin D supplementation. However, none of the intervention studies were specifically designed for evaluating the relation between vitamin D and lipids, none had hyperlipemia as an inclusion criterion, and none were sufficiently powered. In only one study was a significant effect seen with an 8% (0.28 mmol/L) increase in serum LDL-C and a 16% (0.22 mmol/L) decrease in serum TG in those given vitamin D as compared to the placebo group. Accordingly, the effect of vitamin D supplementation on serum lipids is at present uncertain. Considering the numerous other promising vitamins and minerals that when properly tested have been disappointing, one should wait for the results of forthcoming vitamin D intervention studies before drawing conclusions on potential beneficial effects of vitamin D. PMID- 21640758 TI - Loop-mediated isothermal amplification for rapid detection of Chinese sacbrood virus. AB - Chinese sacbrood virus (CSBV) has emerged as an important etiologic agent of honeybee infections and is lethal for individual bees, even causing the collapse of entire colonies. Although diagnostic methods for CSBV have been established in many clinical laboratories, application of these methods is largely restricted by the apparatus needed to carry out the reaction and by cost, therefore a simpler and less expensive diagnostic method for CSBV infection is required. In this study a simple and inexpensive system is described that is based on the loop mediated isothermal amplification (LAMP) assay. The LAMP and the polymerase chain reaction (PCR) methods were compared for their ability to detect CSBV in 31 clinical samples, in purified CSBV-LNQY strains or to be able to discriminate between cDNA samples from other viruses. The detection limit of the LAMP method was 1pg, showing that LAMP is as sensitive as reverse transcriptase (RT)-PCR for CSBV detection. In addition, no DNA band from other related viruses samples was amplified by either method, suggesting that this LAMP assay is as specific as RT PCR for CSBV detection. All 31 clinical samples that were LAMP assay-positive were also amplified by RT-PCR, however the LAMP assay was faster, more cost effective, and easier to perform as the target gene amplified rapidly, within 2h, and only a standard laboratory water bath or heat block was required for the reaction. The results demonstrate clearly that this LAMP-based assay is a useful tool for the rapid and sensitive diagnosis of CSBV infection of bees. PMID- 21640759 TI - Hepatoprotective effect of ghrelin on carbon tetrachloride-induced acute liver injury in rats. AB - BACKGROUND & AIMS: Recent studies have revealed that ghrelin may be an antioxidant and antiinflammatory agent. Oxidative stress are considered to play a prominent causative role in the development of various hepatic disorders. We investigated whether ghrelin plays a protective role against carbon tetrachloride (CCl(4))-induced acute liver injury in rats. METHODS: Forty adult male Sprague Dawley rats were randomly divided into four equal groups as; control, ghrelin, CCl(4) and ghrelin plus CCl(4). Evaluations were made for lipid peroxidation, enzyme activities and biochemical parameters. Pathological histology was also performed. RESULTS: CCl(4) treatment increased plasma and liver tissue malondialdehyde (MDA) content and plasma nitric oxide (NO) level, and decreased erythrocyte and liver tissue superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) activities when compared to control group. At the same time, CCl(4) treatment increased the serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alcaline phosphatase (ALP) activities. By contrast, ghrelin pretreatment reduced plasma and liver MDA content and plasma NO level, and increased erythrocyte and liver tissue SOD, CAT and GPx activities when compared with CCl(4)-treated group. Moreover, both ghrelin alone and ghrelin plus CCl(4) treatment elevated serum glucose level. The CCl(4)-induced histopathological changes were also reduced by the ghrelin pretreatment. CONCLUSION: Our results show that ghrelin can be proposed to protect the liver against CCl(4)-induced oxidative damage in rats, and the hepatoprotective effect may be correlated with its antioxidant and free radical scavenger effects. PMID- 21640760 TI - Inhibitory effect of somatostatin on the NPY response to insulin-induced hypoglycemia and the role of endogenous opioids. AB - The present study was undertaken in order to establish whether somatostatin (SRIH) is able to modify the neuropeptide Y (NPY) response to insulin-induced hypoglycemia during insulin tolerance test (ITT) in man. In addition, the possible involvement of opioid peptides in the mediation of hypoglycemia and/or SRIH action was investigated. Subjects were injected intravenously with 0.15IU/kg insulin alone (control test) or with SRIH (4.1MUg/min/90min), naloxone (10mg in an iv bolus) or the combination of the two substances. Plasma NPY concentrations rose significantly during ITT. The NPY response was significantly reduced by the treatment with SRIH. The administration of naloxone did not modify NPY levels whereas when both SRIH and naloxone were given, NPY response to hypoglycemia did not differ from that observed in the control test. These data demonstrate that SRIH inhibits the NPY response to hypoglycemia. Naloxone-sensitive endogenous opiates do not seem to be involved in the control of hypoglycemia-induced NPY release. In contrast, since naloxone reversed the inhibiting effect of SRIH, an involvement of opioid peptides in the SRIH action may be supposed. PMID- 21640761 TI - Potential role of intermedin/adrenomedullin 2 in early embryonic development in rats. AB - Adrenomedullin2 (ADM2), also referred to as Intermedin (IMD) is expressed in trophoblast cells in human placenta and enhances the invasion and migration of first trimester HTR-8/SV-neo cells. Recently we demonstrated that infusion of IMD antagonist in pregnant rats causes feto-placental growth restriction suggesting a role for IMD in maintaining a successful pregnancy. Therefore, this study was undertaken to assess if IMD has a functional role in embryo implantation in a rat model. We show that IMD mRNA is expressed in rat implantation sites and its expression is significantly higher on day 15 in placenta compared to days 18-22. Infusion of IMD antagonist IMD17-47 from day 3 of pregnancy causes a significant decrease in the weights of day 9 implantation sites as well as serum levels of 17beta-estradiol, progesterone, nitric oxide and serum MMP2 and MMP9 gelatinase activity. Further, expression of MMP2, MMP9, VEGF and PLGF protein levels are significantly downregulated in the implantation sites of IMD antagonist treated rats. This study suggests a potential involvement of IMD in regulating the factors that are critical for implantation and growth of the embryo and thus in establishment of normal rat pregnancy. PMID- 21640762 TI - Impact of atorvastatin on serum vaspin levels in hypercholesterolemic patients with moderate cardiovascular risk. AB - AIM: Vaspin and visfatin have emerged as novel adipokines, involved in atherosclerosis progression. The aim of the present study was to investigate the effects of atorvastatin and lifestyle modification on the above adipokines in hypercholesterolemic patients. METHODS: One hundred four statin-free subjects with moderate cardiovascular risk (Framingham risk score of 10-20%) were randomly assigned to receive either atorvastatin 20mg per day (AT group, n=52) or lifestyle modification (LM, group, n=52). For comparison, age and gender-matched blood donors, without any chronic cardiovascular or metabolic disease served as healthy controls (HC group, n=40). Clinical and anthropometrical parameters, lipids, fasting glucose, serum vaspin, visfatin and insulin levels were obtained at the beginning and after 12 weeks. RESULTS: At the end of the study, intra group and inter-group comparison revealed that atorvastatin administration considerably ameliorated most lipid parameters and downregulated hsCRP levels (p=0.002, p=0.041, respectively). Moreover, we observed a significant increase of vaspin concentrations after 12-week atorvastatin treatment (from 1.37+/-0.6ng/ml to 2.13+/-0.61ng/ml), as compared to baseline (p=0.007) and LM group (p=0.030). In standard multiple regression analysis, the atorvastatin-induced decrease of vaspin was independently associated with hsCRP reduction (p=0.015). On the other hand, within and between groups comparison revealed a non-significant (p>0.05) reduction of visfatin serum levels. In our study, lifestyle modification had totally modest influence on clinical and biochemical variables (p>0.05). CONCLUSION: In hypercholesterolemic patients with moderate estimated cardiovascular risk, atorvastatin administration reduced hsCRP and increased serum vaspin levels compared to lifestyle modification. The relation of those pleiotropic, non-lipid-lowering effects of statins with their clinical outcomes remains to be proved. PMID- 21640763 TI - alphaB-crystallin, a small heat shock protein, modulates NF-kappaB activity in a phosphorylation-dependent manner and protects muscle myoblasts from TNF-alpha induced cytotoxicity. AB - alphaB-crystallin, a member of the small heat shock protein family, has been implicated in various biological functions including response to heat shock, differentiation and apoptosis, the mechanisms of which have not been well understood. Myoblasts, the precursor cells in muscle regeneration, when subjected to growth factor deprivation differentiate to form myotubes or undergo apoptosis. During differentiation, myoblasts express elevated levels of alphaB-crystallin as well as TNF-alpha but the connecting link between these proteins in cell signaling is not clearly understood. We have therefore investigated the role of alphaB-crystallin in TNF-alpha induced regulation of NF-kappaB. We demonstrate that in response to TNF-alpha treatment, alphaB-crystallin associates with IKKbeta and activate its kinase activity, facilitating the degradation of phosphorylated I-kBalpha, a prime step in NF-kappaB activation. Reducing the level of alphaB-crystallin using the RNAi approach reduces the translocation of p65, further confirming the role of alphaB-crystallin in NF-kappaB activation. Our study shows that the ability of alphaB-crystallin to activate NF-kappaB depends on its phosphorylation status. The present study shows that alphaB crystallin-dependent NF-kappaB activation protects myoblasts from TNF-alpha induced cytoxicity by enhancing the expression of the anti-apoptotic protein, Bcl 2. Thus, our study identifies yet another mechanism by which alphaB-crystallin exerts its anti-apoptotic activity. PMID- 21640764 TI - Cannabinoid receptor 2 undergoes Rab5-mediated internalization and recycles via a Rab11-dependent pathway. AB - Cannabinoid receptor 2 (CB2) is a GPCR highly expressed on the surface of cells of the immune system, supporting its role in immunomodulation. This study has investigated the trafficking properties of this receptor when stably expressed by HEK-293 cells. As previously reported, cell surface CB2 rapidly internalized upon exposure to agonist. Direct evidence of CB2 recycling was observed upon competitive removal of the stimulating agonist by inverse agonist. CB2 also underwent slow constitutive internalization when agonist was absent and was up regulated in the presence of inverse agonist. Co-expression of CB2 and dominant negative Rab5 resulted in a significantly reduced capacity for receptors to internalize with no effect on recycling of the internalized receptors. Conversely, co-expression with dominant negative Rab11 did not alter the ability of CB2 to internalize but did impair their ability to return to the cell surface. Co-expression of wild-type, dominant negative or constitutively active Rab4 with CB2 did not alter basal surface expression, extent of internalization, or extent of recycling. These results suggest that Rab5 is involved in CB2 endocytosis and that internalized receptors are recycled via a Rab11 associated pathway rather than the rapid Rab4 associated pathway. This report provides the first comprehensive description of CB2 internalization and recycling to date. PMID- 21640765 TI - Development of an extended multilocus sequence typing for genotyping of Brucella isolates. AB - By amplifying and sequencing longer sequences, an extended multi locus sequence typing (EMLST) theme was developed for Brucella. 61 isolates were genotyped by the EMLST with increased resolution. This strategy could be extended to other bacteria to improve MLST genotyping resolution without additional loci. PMID- 21640766 TI - Event-related potentials in children with attention deficit hyperactivity disorder: an investigation using an auditory oddball task. AB - The aim of the study was to investigate differences in electrophysiological brain activity between children diagnosed with attention deficit hyperactivity disorder combined type (ADHD-Com) and normally developing children, using the auditory 2 tone oddball paradigm. Forty right-handed subjects aged between 6.9 and 12.3 years participated in the present study, with 16 boys and 4 girls in each of the control and ADHD-Com groups. Children were individually age- and gender-matched. The auditory oddball task consisted of 155 standards (1 KHz, p=.66) and 80 targets (1.5 KHz, p=.34), presented randomly one at a time. Subjects were instructed to listen to the sounds and count the rare tones. Task performance in ADHD children did not differ compared to that in the control group. Event-related potentials (ERPs) elicited to target and standard stimuli were analyzed for between-group differences. The ADHD group showed enhanced P2 and reduced N2 component to both oddball stimuli, followed by reduced P3 component to attended targets compared with controls. The difference in the P3 amplitude between targets and standards was smaller in the ADHD group, particularly over the right hemisphere. These results suggest deficiencies in both automatic and controlled processing in children with ADHD. Enhanced amplitude of the P2 in ADHD children may reflect an early orienting deficit which affects later processing stages in the oddball task. Reduced amplitude of the N2 in the clinical group may be associated with stimulus discrimination impairment and inappropriate conflict monitoring. Reduced amplitude of target P3 and its asymmetrical distribution in ADHD children may reflect a deficit in higher-level executive functions, such as attention allocation and stimulus evaluation, accompanied by an impairment of global aspects of attentional processing that are under right hemisphere control. PMID- 21640767 TI - Comparison of piggyBac transposition efficiency between linear and circular donor vectors in mammalian cells. AB - The piggyBac transposon has recently attracted attention as a tool for transgene integration in mammalian cells. However, previous studies involving piggyBac investigated only transposition from circular DNA, although some linear DNA vectors are used to transfect mammalian cells. In this study, we compared the transposition efficiency of piggyBac between linear and circular DNA. Colony counting assay, luciferase assay, and plasmid rescue assay showed that piggyBac transposon can transpose from linear DNA, but its efficiency is lower than the transposition efficiency from circular DNA. These results suggest that circular DNA is more suitable as donor vectors of piggyBac than linear DNA. PMID- 21640768 TI - Mechanism of lipid extraction from Botryococcus braunii FACHB 357 in a biphasic bioreactor. AB - Algal lipid of Botryococcus braunii could be produced continuously and in situ extracted in an aqueous-organic bioreactor. In this study, the cell ultra structure and cell membrane permeability of B. braunii FACHB 357 were investigated to understand the mechanism of lipid extraction within the biphasic system. The results showed that biocompatible solvent of tetradecane could induce algal lipid accumulation, enable the cell membrane more active and the cell wall much looser. The exocytosis process was observed to be one of the mechanisms for lipid cross-membrane extraction in the presence of organic solvent. PMID- 21640769 TI - Establishment of an Atlantic salmon kidney cell line with an inducible gene expression system. AB - A stable recombinant Atlantic Salmon Kidney cell line ASK for use as an inducible expression system was isolated, cloned and characterised. The cells were transfected with the pTet-Off plasmid from the Tet On/Off Clontech system, carrying a G418 resistance gene. Several G418-resistant clones were sub-cultured and characterised by qPCR and by transient transfection. The level of expression of transcriptional activator (tTA) was measured by qPCR in a number of isolated clones. Transient transfection with a pTRE2-hyg-LUC plasmid was used to evaluate the inducibility of these clones. Two clones were chosen for their compromise between cell growth and inducibility. This genetically engineered cell line is a valuable tool for the fish research community especially in research areas investigating the biological function of viral proteins. PMID- 21640770 TI - Hypermethylated levels of E-cadherin promoter in Huh-7 cells expressing the HCV core protein. AB - BACKGROUND AND AIM: The mechanisms of hepatocarcinogenesis induced by hepatitis C virus remain unclear. Our aim was to investigate the effect of the HCV core protein on the promoter methylation status of selected genes potentially involved in the hepatocellular carcinoma (HCC). MATERIALS AND METHODS: We evaluated the promoter methylation levels of the E-cadherin (CDH1), the glutathione S transferase p1 (GSTP1), adenomatosis polyposis coli (APC), tissue inhibitor of metalloproteinase 3 (TIMP3), catenin (cadherin-associated protein) beta 1 (CNNTB1) genes by a quantitative methylation-specific polymerase chain reaction (QMSP) in the in vitro model of Huh-7 cells expressing the HCV core protein of genotype 1b. RESULTS: We found that CDH1 promoter was hypermethylated in genotype 1b HCV core protein-positive cells as compared to control cells expressing the GFP protein alone (HCV core 1b vs GFP p=0.00; HCV core 1b vs Huh-7 p=0.03). This resulted in reduced levels of CDH1 protein as evaluated by immunoblot and by immunofluorescence. On the other hand no significant changes were observed for the other genes investigated. Furthermore, we present evidence that genotype 1b HCV core protein expression induces SIRT1 upregulation and that treatment with SIRT1 inhibitor sirtinol decreases the methylation levels of CDH1 promoter (1b+sirtinol vs 1b p=0.05; 1b+sirtinol vs GFP+sirtinol p=NS) resulting in 1.7 fold increased CDH1 mRNA expression (1b+sirtinol vs 1b p=0.05). CONCLUSIONS: Our findings suggest that HCV core protein could play a role in HCC at least in part by altering the methylation status of CDH1 promoter. These findings could also suggest a novel therapeutic approach for HCC. PMID- 21640771 TI - Immunophilin-loaded erythrocytes as a new delivery strategy for immunosuppressive drugs. AB - Cyclosporine A (CsA) and tacrolimus (also known as FK506) are natural compounds with immunosuppressive activity that have improved the outcome of organ transplantation. Unfortunately, both drugs are characterised by high pharmacokinetic variability, poor bioavailability and high toxicity. Until now, no optimal method to deliver immunosuppressant drugs into circulation has been developed. Here we propose the use of engineered erythrocytes as a drug delivery system for the release of immunosuppressants in circulation in order to modify their pharmacokinetic and restrain toxic effects. After administration, FK506 and CsA mainly distribute within erythrocytes owing to the presence into these cells of immunophilins that bind the drugs with very high affinity (FKBP12 for FK506 and cyclophilin A for CsA); therefore, a new strategy aimed to increase the amount of FK506/CsA carried by erythrocytes by increasing the intra-erythrocytic concentration of the respective binding proteins has been developed. We manufactured recombinant forms of human FKBP12 and cyclophilin A to be loaded into RBC through a hypotonic dialysis and isotonic resealing procedure. Erythrocytes loaded with 3.5+/-1.3, 7.5+/-3.1 and 15.5+/-0.4nmol FKBP12 were able to bind 3.5+/-1.5, 6.0+/-1.9 and 11.4+/-2.9MUg FK506 per millilitre RBC, respectively, while RBC loaded with 4.0+/-0.6, 5.0+/-0.8 and 15.9+/-2.4nmol of cyclophilin A could bind 8.9+/-3.4, 12.2+/-3.5 and 17.0+/-3.2MUg CsA. Thus, both engineered RBC were demonstrated able to bind up to an order of magnitude more drug than corresponding native erythrocytes (1.0+/-0.3MUg FK506 and 3.2+/-0.3MUg CsA). Moreover, FK506 released from FKBP12-RBC is able to be up-taken by T lymphocytes and inhibit IL-2 expression in vitro as free administered drug. In summary, our results indicate that diffusible immunosuppressants could be entrapped into red cells (thanks to the loading of the respective target protein) and suggest that immunophilin-loaded RBC could be employed as potential delivery system for immunosuppressive agents. PMID- 21640772 TI - In silico models of aerosol delivery to the respiratory tract - development and applications. AB - This review discusses the application of computational models to simulate the transport and deposition of inhaled pharmaceutical aerosols from the site of particle or droplet formation to deposition within the respiratory tract. Traditional one-dimensional (1-D) whole-lung models are discussed briefly followed by a more in-depth review of three-dimensional (3-D) computational fluid dynamics (CFD) simulations. The review of CFD models is organized into sections covering transport and deposition within the inhaler device, the extrathoracic (oral and nasal) region, conducting airways, and alveolar space. For each section, a general review of significant contributions and advancements in the area of simulating pharmaceutical aerosols is provided followed by a more in depth application or case study that highlights the challenges, utility, and benefits of in silico models. Specific applications presented include the optimization of an existing spray inhaler, development of charge-targeted delivery, specification of conditions for optimal nasal delivery, analysis of a new condensational delivery approach, and an evaluation of targeted delivery using magnetic aerosols. The review concludes with recommendations on the need for more refined model validations, use of a concurrent experimental and CFD approach for developing aerosol delivery systems, and development of a stochastic individual path (SIP) model of aerosol transport and deposition throughout the respiratory tract. PMID- 21640773 TI - The shaping role of hunger on self-reported external eating status. AB - As people are relatively incompetent in assessing the impact of visceral states on their behavior, two studies tested the hypothesis that hunger affects the extent to which people assess themselves as external eaters. In Study 1 participants' current self-reported hunger states were linked to their scores on an external eating scale. Hungrier participants perceived themselves more strongly as external eaters. In Study 2 hunger was experimentally manipulated, after which self-reported external eating was assessed. Hunger was found to affect people's self-reported external eating status, such that hungry participants scored higher and above the average norm score on external eating compared to satiated participants, who scored below this average norm score. The key implications of these findings are discussed. PMID- 21640774 TI - Effects of increased hypothalamic leptin gene expression on ovariectomy-induced bone loss in rats. AB - Estrogen deficiency results in accelerated bone turnover with a net increase in bone resorption. Subcutaneous administration of leptin attenuates bone loss in ovariectomized (ovx) rats by reducing bone resorption. However, in addition to its direct beneficial effects, leptin has been reported to have indirect (central nervous system-mediated) antiosteogenic effects on bone, which may limit the efficacy of elevated serum leptin to prevent estrogen deficiency-associated bone loss. The present study evaluated the long-term effects of increased hypothalamic leptin transgene expression, using recombinant adeno-associated virus-leptin (rAAV-Lep) gene therapy, on bone mass, architecture, and cellular endpoints in sexually mature ovx Sprague-Dawley rats. Ovx rats were implanted with cannulae in the 3rd ventricle of the hypothalamus and injected with either rAAV-Lep or rAAV GFP (control vector encoding green fluorescent protein) and maintained for 10 weeks. Additional controls consisted of ovary-intact rats and ovx rats pair-fed to rAAV-Lep rats. Lumbar vertebrae were analyzed by micro-computed tomography and tibiae by histomorphometry. Cancellous bone volume was lower and osteoclast perimeter, osteoblast perimeter, and bone marrow adipocyte density were greater in ovx rats compared to ovary-intact controls. In contrast, differences among ovx groups were not detected for any endpoint evaluated. In conclusion, whereas estrogen deficiency resulted in marked cancellous osteopenia, increased bone turnover and marrow adiposity, increasing hypothalamic leptin transgene expression in ovx rats had neither detrimental nor beneficial effects on bone mass, architecture, or cellular endpoints. These findings demonstrate that the antiresorptive effects of subcutaneous leptin administration in ovx rats are mediated through leptin targets in the periphery. PMID- 21640776 TI - Lack of correlation between BCG-induced tuberculin skin test sensitisation and protective immunity in cattle. AB - Vaccination of cattle with Mycobacterium bovis Bacille Calmette-Guerin (BCG) can provide significant protection against bovine tuberculosis (TB). However, BCG vaccination sensitises animals to respond to the tuberculin skin-test. This provides a potential operational impediment to the use of BCG as a cattle vaccine since the tuberculin skin-test is the primary surveillance tool used by many countries with 'test and slaughter' control strategies. Currently, it is also unclear what BCG-induced skin-test conversion means in respects to BCG's protective immunity. In the current study we first investigated the duration of tuberculin skin-test sensitisation in calves neonatally vaccinated with BCG. BCG vaccination induced strong skin-test responses in calves during their first 6 months. However, a rapid decay in skin-test sensitivity was observed after this time. Between 6 and 9 months this represented a reduction from 80% to 8% of calves providing a positive response in the single intradermal comparative cervical tuberculin test at standard interpretation. We next investigated the relationship between BCG induced skin-test sensitivity and retention of protective immunity. Calves were neonatally vaccinated with BCG and subsequently divided into 2 groups based on retention or loss of tuberculin skin-test responses after 6 months. In contrast to their skin-test responsiveness, these vaccinates maintained their tuberculin specific IFN-gamma blood responses. Moreover, irrespective of their pre-challenge skin-test responses, following M. bovis challenge both groups of BCG vaccinated calves demonstrated comparable levels of protection, as evidenced by reduced TB-associated pathology. Therefore, we have demonstrated that following neonatal BCG vaccination of cattle, tuberculin skin-test responder frequencies waned rapidly after 6 months but importantly, loss of skin-test sensitivity did not correlate with loss of protective immunity. These findings could have implications for the practical application of BCG based cattle vaccines. PMID- 21640775 TI - HPV vaccination among a community sample of young adult women. AB - OBJECTIVES: Despite the high efficacy of the human papillomavirus (HPV) vaccine, uptake has been slow and little data on psychosocial barriers to vaccination exist. METHODS: A community sample of 428 women enrolled in a longitudinal study of social development in the Seattle WA metropolitan area were interviewed about HPV vaccine status, attitudes, and barriers to HPV vaccination in spring 2008 or 2009 at ~age 22. RESULTS: Nineteen percent of women had initiated vaccination, 10% had completed the series, and ~40% of unvaccinated women intended to get vaccinated. Peer approval was associated with vaccine initiation (adjusted prevalence ratio (APR) 2.1; 95% confidence interval 1.4-3.2) and intention to vaccinate (APR 1.4; 1.1-1.9). Belief the vaccine is <75% effective was associated with less initiation (APR 0.6; 0.4-0.9) or intention to vaccinate (APR 0.5; 0.4 0.7). Vaccine initiation was also less likely among cigarette smokers and illegal drug users, whereas intention to vaccinate was more common among women currently attending school or with >5 lifetime sex partners, but less common among women perceiving low susceptibility to HPV (APR 0.6; 0.5-0.9). CONCLUSIONS: HPV vaccination uptake was low in this community sample of young adult women. Increasing awareness of susceptibility to HPV and the high efficacy of the vaccine, along with peer interventions to increase acceptability, may be most effective. PMID- 21640777 TI - Reducing animal experimentation in foot-and-mouth disease vaccine potency tests. AB - The World Organisation for Animal Health (OIE) Terrestrial Manual and the European Pharmacopoeia (EP) still prescribe live challenge experiments for foot and-mouth disease virus (FMDV) immunogenicity and vaccine potency tests. However, the EP allows for other validated tests for the latter, and specifically in vitro tests if a "satisfactory pass level" has been determined; serological replacements are also currently in use in South America. Much research has therefore focused on validating both ex vivo and in vitro tests to replace live challenge. However, insufficient attention has been given to the sensitivity and specificity of the "gold standard"in vivo test being replaced, despite this information being critical to determining what should be required of its replacement. This paper aims to redress this imbalance by examining the current live challenge tests and their associated statistics and determining the confidence that we can have in them, thereby setting a standard for candidate replacements. It determines that the statistics associated with the current EP PD(50) test are inappropriate given our domain knowledge, but that the OIE test statistics are satisfactory. However, it has also identified a new set of live animal challenge test regimes that provide similar sensitivity and specificity to all of the currently used OIE tests using fewer animals (16 including controls), and can also provide further savings in live animal experiments in exchange for small reductions in sensitivity and specificity. PMID- 21640779 TI - Mutual interference on the immune response to yellow fever vaccine and a combined vaccine against measles, mumps and rubella. AB - A randomized trial was conducted to assess the immunogenicity and reactogenicity of yellow fever vaccines (YFV) given either simultaneously in separate injections, or 30 days or more after a combined measles-mumps-rubella (MMR) vaccine. Volunteers were also randomized to YFV produced from 17DD and WHO-17D 213 substrains. The study group comprised 1769 healthy 12-month-old children brought to health care centers in Brasilia for routine vaccination. The reactogenicity was of the type and frequency expected for the vaccines and no severe adverse event was associated to either vaccine. Seroconversion and seropositivity 30 days or more after vaccination against yellow fever was similar across groups defined by YFV substrain. Subjects injected YFV and MMR simultaneously had lower seroconversion rates--90% for rubella, 70% for yellow fever and 61% for mumps--compared with those vaccinated 30 days apart--97% for rubella, 87% for yellow fever and 71% for mumps. Seroconversion rates for measles were higher than 98% in both comparison groups. Geometric mean titers for rubella and for yellow fever were approximately three times higher among those who got the vaccines 30 days apart. For measles and mumps antibodies GMTs were similar across groups. MMR's interference in immune response of YFV and YFV's interference in immune response of rubella and mumps components of MMR had never been reported before but are consistent with previous observations from other live vaccines. These results may affect the recommendations regarding primary vaccination with yellow fever vaccine and MMR. PMID- 21640778 TI - Intranasal delivery of Norwalk virus-like particles formulated in an in situ gelling, dry powder vaccine. AB - The development of a vaccine to prevent norovirus infections has been focused on immunization at a mucosal surface, but has been limited by the low immunogenicity of self-assembling Norwalk virus-like particles (NV VLPs) delivered enterically or at nasal surfaces. Nasal immunization, which offers the advantage of ease of immunization, faces obstacles imposed by the normal process of mucociliary clearance, which limits residence time of applied antigens. Herein, we describe the use of a dry powder formulation (GelVac) of an inert in situ gelling polysaccharide (GelSite) extracted from Aloe vera for nasal delivery of NV VLP antigen. Powder formulations, with or without NV VLP antigen, were similar in structure in dry form or when rehydrated in simulated nasal fluids. Immunogenicity of the dry powder VLP formulation was compared to equivalent antigen/adjuvant liquid formulations in animals. For the GelVac powder, we observed superior NV-specific serum and mucosal (aerodigestive and reproductive tracts) antibody responses relative to liquid formulations. Incorporation of the TLR7 agonist gardiquimod in dry powder formulations did not enhance antibody responses, although its inclusion in liquid formulations did enhance VLP immunogenicity irrespective of the presence or absence of GelSite. We interpret these data as showing that GelSite-based dry powder formulations (1) stabilize the immunogenic structural properties of VLPs and (2) induce systemic and mucosal antibody titers which are equal or greater than those achieved by VLPs plus adjuvant in a liquid formulation. We conclude that in situ gelation of the GelVac dry powder formulation at nasal mucosal surfaces delays mucociliary clearance and thereby prolongs VLP antigen exposure to immune effector sites. PMID- 21640780 TI - Efficacy, safety and immunogenicity of RIX4414 in Japanese infants during the first two years of life. AB - A phase III, randomized, double-blind study evaluated the efficacy, reactogenicity, safety and immunogenicity of a human rotavirus vaccine, RIX4414 in Japanese infants aged 6-14 weeks when administered as two doses (0, 1-month schedule). Efficacy against any and severe rotavirus gastroenteritis leading to medical intervention caused by circulating wild-type rotavirus from two weeks post-Dose 2 until two years of age was 79.3% (95% CI: 60.5-89.8%) and 91.6% (95% CI: 62.4-99.1%), respectively. Solicited, unsolicited symptoms and serious adverse events were reported at a similar frequency in both groups. Serum anti rotavirus antibody seroconversion rate one-month post-Dose 2 was 85.3% (95% CI: 68.9-95%) in RIXX4414 group. RIX4414 was efficacious, well-tolerated and immunogenic in Japanese infants and introduction of vaccination could help in reducing the disease burden. PMID- 21640781 TI - Introduction to Bayesian statistical approaches to compositional analyses of transgenic crops 1. Model validation and setting the stage. AB - Statistical comparisons of compositional data generated on genetically modified (GM) crops and their near-isogenic conventional (non-GM) counterparts typically rely on classical significance testing. This manuscript presents an introduction to Bayesian methods for compositional analysis along with recommendations for model validation. The approach is illustrated using protein and fat data from two herbicide tolerant GM soybeans (MON87708 and MON87708*MON89788) and a conventional comparator grown in the US in 2008 and 2009. Guidelines recommended by the US Food and Drug Administration (FDA) in conducting Bayesian analyses of clinical studies on medical devices were followed. This study is the first Bayesian approach to GM and non-GM compositional comparisons. The evaluation presented here supports a conclusion that a Bayesian approach to analyzing compositional data can provide meaningful and interpretable results. We further describe the importance of method validation and approaches to model checking if Bayesian approaches to compositional data analysis are to be considered viable by scientists involved in GM research and regulation. PMID- 21640782 TI - Zolpidem reduces the blood oxygen level-dependent signal during visual system stimulation. AB - Zolpidem is a short-acting imidazopyridine hypnotic that binds at the benzodiazepine binding site on specific GABA(A) receptors to enhance fast inhibitory neurotransmission. The behavioral and receptor pharmacology of zolpidem has been studied extensively, but little is known about its neuronal substrates in vivo. In the present within-subject, double-blind, and placebo controlled study, blood oxygen level-dependent functional magnetic resonance imaging (BOLD fMRI) at 3 Tesla was used to assess the effects of zolpidem within the brain. Healthy participants (n=12) were scanned 60 min after acute oral administration of zolpidem (0, 5, 10, or 20mg), and changes in BOLD signal were measured in the visual cortex during presentation of a flashing checkerboard. Heart rate and oxygen saturation were monitored continuously throughout the session. Zolpidem (10 and 20mg) reduced the robust visual system activation produced by presentation of this stimulus, but had no effects on physiological activity during the fMRI scan. Zolpidem's modulation of the BOLD signal within the visual cortex is consistent with the abundant distribution of GABA(A) receptors localized in this region, as well as previous studies showing a relationship between increased GABA-mediated neuronal inhibition and a reduction in BOLD activation. PMID- 21640783 TI - The correlation of plasma Abeta42 levels, depressive symptoms, and cognitive function in the Korean elderly. AB - OBJECTIVES: This study aims to investigate whether plasma amyloid beta 1-42 (Abeta42) levels are associated with depressive symptoms and/or cognitive function in community dwelling elderly. METHODS: Subjects were 123 participants of a population-based project designed to screen community dwelling elderly older than 65 years old in Gangwon Province, Korea, for the early detection of depression and dementia. Symptoms of depression were assessed using the SGDS-K (Short Geriatric Depression Scale-Korean version), and the MMSE-KC (Mini-Mental State Examination-Korean version) was used to assess cognitive function. Plasma Abeta42 levels were measured with the human amyloid beta ELISA Kit. RESULTS: The elderly with depressive symptoms (SGDS-K score >= 8) had higher plasma Abeta42 levels than those without depressive symptoms (SGDS-K score<8) (p<0.1). Plasma Abeta42 levels were positively correlated with SGDS-K scores (p<0.05). However, MMSE-KC scores were inversely associated with plasma Abeta42 levels (p<0.01). Plasma beta42 levels were also associated with MMSE-KC (F=8.07, p<0.01) and SGDS K (F=4.53, p<0.05) by generalized linear model (GLM) with controlling age, sex and education. CONCLUSION: Plasma Abeta42 levels were associated with depressive symptoms and cognitive function in community dwelling elderly. The present study supports the possibility that plasma Abeta may be involved in the development of late onset depression. PMID- 21640784 TI - Possible role of cysteine-S-conjugate beta-lyase in species differences in cisplatin nephrotoxicity. AB - To better understand species differences in cisplatin nephrotoxicity, we focused on renal cysteine-S-conjugate beta-lyase (C-S lyase), which may play a crucial role in the metabolism of platinum (Pt)-cysteine conjugates. Aminooxyacetic acid hemihydrochloride (AOAA), an inhibitor of C-S lyase, reduced renal injuries due to cisplatin in rats, suggesting involvement of C-S lyase. On day 5 following a bolus cisplatin injection, three species showed in vivo nephrotoxic potentials in the order of rats>mice=rabbits (the highest to lowest), based on body surface. The levels of renal Pt residue at the nephrotoxic dose were in order of rabbits>rats>mice. Meanwhile, the activity of endogenous (basal) mitochondrial aspartate aminotransferase (AST), one of the C-S lyases, in the renal cortex of naive animals was rats>mice=rabbits. In a qualitative Western blot analysis, expression of mitochondrial C-S lyase in the kidney was observed at approximately 37kDa in all five species used. In in vitro studies, the cytotoxicity of cisplatin was dependent on the expression level of C-S lyase mRNA in the respective renal cells. These results demonstrate that species differences in cisplatin nephrotoxicity are attributable to an interaction of renal Pt transition with C-S lyase activity. PMID- 21640785 TI - Comparative cytotoxicity study of enniatins A, A1, A2, B, B1, B4 and J3 on Caco-2 cells, Hep-G2 and HT-29. AB - Enniatins (ENs) are ionophoric, phytotoxic, antihelminthic, and antibiotic compounds of hexadepsipeptidic structure produced by several strains of Fusarium spp. The cytotoxicity effect of the ENs A, A(1), A(2), B, B(1), B(4) and J(3) was compared on three tumor cell lines, the human epithelial colorectal adenocarcinoma (Caco-2), the human colon carcinoma (HT-29), and the human liver carcinoma (Hep-G2). The endpoint evaluated was the mitochondrial integrity by using the MTT assays, after 24 and 48 h of incubation. The IC(50) value for EN A(2) on Caco-2 cells, after 24h exposure, was 18.7 +/- 4.5 MUM and decrease to 2.6 +/- 0.7 MUM at 48 h of incubation. However, ENs A, A(1), B(1) and B(4) exert pronounced cytotoxic effects in all the cell lines tested by the MTT assay after 24 and 48 h of incubation. The EN A(1) demonstrated to be the most cytotoxic ENs tested. Moreover, no statistical differences were found between the IC(50) values obtained for EN A(1) on Caco-2, HT-29 and Hep-G2, with IC(50) values ranging from 9.1 +/- 2.2 MUM to 12.3 +/- 4.3 MUM at 24h and decreasing in a range variable from 1.4 +/- 0.7 MUM to 2.7 +/- 0.8 MUM at 48 h. On the other hand, EN A, B(1) and B(4) showed lower cytotoxicity, but in a similar range as the IC(50) values reported on HT-29 (IC(50) values (24h): 16.8 +/- 4.3-26.2 +/- 6.7 MUM), Caco-2 (IC(50) values (24h): 19.5 +/- 4.1 MUM) and Hep-G2 (IC(50) values (24h): 23.4 +/- 5.6-26.2 +/- 7.6 MUM) cells. Cytotoxic effect with a 48 h of incubation revealed also a significant toxicity of ENs A (IC(50) values ranged from 8.2 +/- 1.8 to 11.4 +/- 4.6 MUM), B(1) (IC(50) values variables from 3.7 +/- 0.7 to 11.5 +/- 5.3 MUM) and B(4) (IC(50) of 4.5 +/- 2.9-15.0 +/- 4.0 MUM). In summary, this study demonstrated that ENs can exert toxic activity at low micromolar concentrations in mammalian cells. PMID- 21640786 TI - Learning to fear obstructed breathing: comparing interoceptive and exteroceptive cues. AB - The present study investigated interoceptive fear conditioning (IFC) to an interoceptive and exteroceptive conditional stimulus (CS) with a severe respiratory load applied for 30s as the unconditional stimulus (US). CSs were another, weak respiratory load in the intero-IFC study (N=74), and a neutral picture in the extero-IFC study (N=42). CSs preceded the US in the paired groups, whereas the unpaired groups received the same number of unpaired CSs and USs. We measured startle blink EMG, self-reported fear and respiration. In the intero-IFC study, the CS-load was associated with larger startle blinks and a smaller decrease in respiratory rate and tidal volume in the paired compared to the unpaired group. In the extero-IFC study, the CS-picture evoked an increase in tidal volume and self-reported fear only in the paired group. In addition, startle potentiation during the CS-picture was greater for the paired than for the unpaired group. PMID- 21640788 TI - Phase-amplitude coupling between theta and gamma oscillations during nociception in rat electroencephalography. AB - In electroencephalography (EEG) study, gamma oscillations were reported to participate in pain processing; theta oscillations were also involved in pain processing. Moreover, theta always modulated gamma activity by phase-amplitude coupling in event-related oscillations. Whether theta modulate gamma by phase amplitude coupling in pain processing is of interest. In the present study, using EEG of rats after laser nociceptive stimulation, we investigated gamma activity and phase-amplitude coupling between theta and gamma. It was found that induced gamma power increased starting 200 ms after nociceptive stimulation onset. Moreover, significant coupling between theta phase and gamma amplitude was found over frontal and parietal region after nociceptive stimulation. Our results for the first time suggest that coupling between theta and gamma is involved in nociception processing. PMID- 21640787 TI - Low-frequency H-reflex depression in trained human soleus after spinal cord injury. AB - After spinal cord injury (SCI), widespread reorganization occurs within spinal reflex systems. Regular muscle activity may influence reorganization of spinal circuitry after SCI. The purpose of this study is to investigate the effects of long-term soleus training on H-reflex depression in humans after SCI. Seven subjects with acute (<7 weeks) SCI (AC group) underwent testing of H-reflex depression at several frequencies of repetitive stimulation. Eight subjects (including 3 from AC) stimulated one soleus muscle daily, leaving the other leg as an untrained within-subject control. Trained limb H-reflexes were assessed during year 1 (TR1) and year 2 (TR2) of training. Untrained limbs were tested during year 2 (UN). H-reflex amplitude was lower at 1, 2 and 5 Hz than at 0.1 or 0.2 Hz (p<0.05). The pattern of depression differed between AC and UN (p<0.05), but not between TR2 and UN (p>0.05) despite significant adaptations in torque and fatigue resistance (p<0.05). Three subjects who began training very early after SCI retained H-reflex post activation depression, suggesting that early intervention of daily muscular activity may be important. PMID- 21640789 TI - Watershed infarcts in a multiple microembolic model of monkey. AB - It has long been debated whether watershed infarcts are caused by hemodynamic or embolic mechanisms. In the present study, we investigated microembolic roles in the pathogenesis of watershed infarcts by examining MRI in a macaque monkey model of multiple microinfarcts. 50 MUm microbeads were injected into each internal carotid artery twice with a month interval. Monkeys (n=4) injected with 2250-2800 microbeads per unilateral side showed both cortical and internal watershed infarcts in the acute phase and atrophic changes with microbleeds in the chronic phase. These results suggest embolic pathogenesis can contribute to the genesis of both cortical and internal watershed infarcts in primates. PMID- 21640790 TI - Differential expression of miRNA-146a-regulated inflammatory genes in human primary neural, astroglial and microglial cells. AB - MicroRNA-146a (miRNA-146a) is an inducible, 22 nucleotide, small RNA over expressed in Alzheimer's disease (AD) brain. Up-regulated miRNA-146a targets several inflammation-related and membrane-associated messenger RNAs (mRNAs), including those encoding complement factor-H (CFH) and the interleukin-1 receptor associated kinase-1 (IRAK-1), resulting in significant decreases in their expression (p<0.05, ANOVA). In this study we assayed miRNA-146a, CFH, IRAK-1 and tetraspanin-12 (TSPAN12), abundances in primary human neuronal-glial (HNG) co cultures, in human astroglial (HAG) and microglial (HMG) cells stressed with Abeta42 peptide and tumor necrosis factor alpha (TNFalpha). The results indicate a consistent inverse relationship between miRNA-146a and CFH, IRAK-1 and TSPAN12 expression levels, and indicate that HNG, HAG and HMG cell types each respond differently to Abeta42-peptide+TNFalpha-triggered stress. While the strongest miRNA-146a-IRAK-1 response was found in HAG cells, the largest miRNA-146a-TSPAN12 response was found in HNG cells, and the most significant miRNA-146a-CFH changes were found in HMG cells, the 'resident scavenging macrophages' of the brain. PMID- 21640791 TI - Proteomic analysis of phosphotyrosyl proteins in human embryonic stem cell derived neural stem cells. AB - Phosphorylation can reveal essential cell functions, such as cell differentiation, signal transduction, metabolic maintenance and cell division. The aim of this study was to investigate phosphorylated protein expression changes during neuronal lineage differentiation from hESCs. To measure the phosphorylated protein expression change during neuronal differentiation, we performed a comparative phosphoproteome analysis using 2-DE after MALDI-TOF MS and an MS/MS protein identification method, making a comparison between neural lineage differentiating cells and normal embryoid bodies (EBs) differentiated from human embryonic stem cells (hESCs) and profiling constituent phosphorylated proteins. Of 36 differentially expressed protein spots, 12 spots were shown to be up-regulated in differentiating neural cells. Specifically, the 7 up-regulated proteins of the 12 have potential roles in neuronal differentiation or neuronal damage recovery, including ACTB, heterogeneous nuclear ribonucleoprotein A2B1 (hnRNP A2B1), heterogeneous nuclear ribonucleoprotein L (hnRNP L), SET, chaperonin-containing TCP-1, vimentin and voltage-dependent anion channel protein 1 (VDAC1). These proteins are discussed further below. PMID- 21640792 TI - Effect of leukemia inhibitory factor on corneal nerve regeneration of rabbit eyes after laser in situ keratomileusis. AB - To explore the effect of leukemia inhibitory factor on corneal nerve regeneration in a rabbit model after laser in situ keratomileusis. Thirty five healthy New Zealand rabbits were divided into three groups for a 6-month observation, the blank control group, the control group, and the treatment group respectively. Laser in situ keratomileusis for myopia was performed on 30 rabbits (60 eyes in total) and then 1 MUg/ml LIF eye drops were used four times a day on the left eyes as the treatment group, and the balanced salt solution (BSS) was used on the right eyes as the control group. Nerve regeneration was evaluated by counting the new regenerated nerves in golden chloride staining. The parameters for dry eye include Schirmer I test and tear break-up time were also examined. The number of regenerated nerve fibers in the treatment group was significantly higher than that in the control group at all time points except the 6th month after LASIK (P<0.05). The parameters for dry eye between two groups were compared at each postoperative time point and the results showed they were significantly higher in the LIF-treated group than in the BSS-control group at 2w, 1m, and 3m respectively. Leukemia inhibitory factor can effectively accelerate the corneal nerve regeneration of rabbit eyes after LASIK surgery and decrease the occurrence of dry eye symptoms. PMID- 21640793 TI - Association between Val66Met polymorphism of Brain-Derived Neurotrophic Factor (BDNF) gene and a deficiency of colour vision in alcohol-dependent male patients. AB - Brain-derived neurotrophic factor (BDNF) is a protein encoded, in humans, by BDNF gene on chromosome 11. BDNF protects adult neurons and promotes growth and differentiation during ontogenetic development but the nature and magnitude of its effects could be influenced by functional polymorphisms. The BDNF polymorphism Val66Met (rs6265) has been studied in the context of etiology of mental diseases including alcoholism. Alcoholism - a complex disorder known to be linked to several genes - has multiple manifestations, including sensory deficits such as those affecting vision. In the present study we examined a relationship between the Val66Met polymorphism, alcohol dependence and colour vision deficiency (CVD) in 167 alcohol-dependent men and 289 control male subjects. Statistical analysis revealed that almost half (about 48%) of the alcohol dependent men had a CVD. In addition we found that CVD was significantly associated (P=0.005) with the Val66Met polymorphism. The A allele containing 66Met promotes BDNF expression and this may protect humans against CVD induced by long-term excessive alcohol intake. The present findings indicate that alcohol induced CVD does not depend solely on excessive alcohol consumption but is significantly influenced by genetic predisposition in the form of a specific BDNF polymorphism. PMID- 21640794 TI - Multi-decade hippocampal and amygdala volume analysis: equal variability and limited age effect. AB - Hippocampal (HC) and amygdala (AG) variability throughout asymptomatic adulthood have not been often characterized. The prevailing assumption is that HC/AG variability is small in young adults, and widens with advancing age and pathology. More recent studies with samples at every decade have reported conflicting results. Our goal was to perform a precise investigation of the effects of Age, Sex and Hemisphere on HC/AG volumes throughout aging. Subjects - we included 422 subjects from the Italian Brain Normative Archive database. Subjects ranged in age from 20 to 84 years. Data - manual segmentation was performed on 422 individuals for the HC, and 228 for the AG, using the Pruessner protocol. Statistical analysis - we tested the influence of total intracranial volume normalization, and used a hierarchical regression model to determine the shape of the association for Age with HC/AG volumes, for both Sex and Hemisphere variables. We explored the distribution of HC/AG volume across age groups by dividing the data into six different strata by decades, and compared volume variability using ANOVA. The study revealed that HC or AG volumes were not significantly related to Age or Age(2), regardless of Sex, except in the right AG. There were no significant differences in variability across age strata. This study lends credence to counter-intuitive notions regarding HC/AG neurodegeneration. Further, researchers can use our HC/AG volumes, broken down by sex and age, as normative data in future fundamental and clinical research. PMID- 21640795 TI - Metallothionein-III prevents neuronal death and prolongs life span in amyotrophic lateral sclerosis model mice. AB - Defect of metallothionein-III (MT-III) has been reported to be a contributor to the progression of amyotrophic lateral sclerosis (ALS). We explored the expression and effects of MT-III on the motor neurons of spinal cords of ALS model mice (G93A Cu/Zn superoxide dismutase (SOD-1) mutant-transgenic (Tg) mice) using a retrograde viral delivery system. Once-weekly injection of the adenovirus encoding LacZ or MT-III gene was started at the age of 20 weeks, which was the mean age of ALS onset. Gene expression was detected in the motor neurons of the lumbar spinal cord. At 160 days of age (14 days after injection), the mean numbers of Nissl-stained alpha neurons were 15.42+/-5.32, 16.50+/-1.35, and 24.75+/-4.01 in 5-MUm sections of the lumbar hemispinal cord from the untreated group, LacZ group, and MT-III group, respectively. The mean durations of illness were 15.20+/-5.30 days, 10.33+/-4.27 days, and 25.71+/-7.67 days in the untreated group, LacZ group, and MT-III group, respectively. The mean life spans were 163.20+/-7.72 days, 159.50+/-3.27 days, and 178.14+/-12.97 days in the untreated group, LacZ group, and MT-III group, respectively. We demonstrated that MT-III prevents the loss of motor neurons of ALS model mice and prolongs the life span, even when the administration is started at the time of onset. PMID- 21640796 TI - Cyclin-dependent kinase 5 with phosphorylation of tyrosine 15 residue is enriched in striatal matrix compartment in adult mice. AB - Accumulating evidence suggests that the striosome-matrix systems have a tight link with motor and behavioral brain functions and their disorders. Cyclin dependent kinase 5 (Cdk5) is a versatile protein kinase that plays a role in synaptic functions and cell survival in adult brain, and its kinase activity is stimulated by phosphorylation at tyrosine 15 residue (pY15). In this study, we used an immunohistochemical method to show differential localization of Cdk5-pY15 in the striatal compartments of adult mice, with a heightened density of Cdk5 pY15 labeling in the matrix relative to the striosomes. Our findings indicate that Cdk5-pY15 can be a new marker for the striatal matrix compartment, and suggest a possible involvement of Cdk5-mediated signaling in compartment-specific neurotransmission and disease pathology in the striatum. PMID- 21640797 TI - Noradrenergic projections to the ventromedial hypothalamus regulate fat metabolism during endurance exercise. AB - The regulation of energy metabolism by the central nervous system during endurance exercise was examined. We conducted respiratory gas analysis by functionally paralyzing the ventromedial hypothalamus (VMH), the lateral hypothalamic area, and the paraventricular nucleus of the hypothalamus with local anaesthetic (lidocaine) during treadmill running at a velocity that allowed for efficient fatty acid oxidation. Our results showed that only the lidocaine treatment of the VMH attenuated fatty acid oxidation during endurance exercise. The monoaminergic neural activities at these nuclei during in vivo microdialysis in rats under the same conditions indicated a significant increase in the extracellular concentration of noradrenaline in all nuclei. Similarly, a significant increase in dopamine occurred at some points during exercise, but no change in serotonin concentration occurred regardless of exercise. Disruption of noradrenergic projections to the VMH by 6-hydroxydopamine attenuated the enhancement of fat oxidation during running. Blocker treatments clarified that noradrenergic inputs to the VMH are mediated by beta-adrenoceptors. These data indicate that information about peripheral tissues status is transmitted via noradrenergic projections originating in the medulla oblongata, which may be an important contribution by the VMH and its downstream mechanisms to enhanced fatty acid oxidation during exercise. PMID- 21640798 TI - Ginkgo biloba extract enhances noncontact erection in rats: the role of dopamine in the paraventricular nucleus and the mesolimbic system. AB - Penile erection is essential for successful copulation in males. Dopaminergic projections from the paraventricular nucleus (PVN) to the ventral tegmental area (VTA) and from the VTA to the nucleus accumbens (NAc) are thought to exert a facilitatory effect on penile erection. Our previous study showed that treatment with an extract of Ginkgo biloba leaves (EGb 761) enhances noncontact erection (NCE) in male rats. However, the relationship between NCE and dopaminergic activity in the PVN, VTA, and NAc remains unknown. The present study examined the relationship between NCE and central dopaminergic activity following EGb 761 treatment. We report here that, in comparison with the controls, there was a significant increase in the number of NCEs in rats after treatment with 50 mg/kg of EGb 761 for 14 days. EGb 761-treated rats also showed more NCEs than the same group before EGb 761 treatment. A significant increase in the expression of catecholaminergic neurons in the PVN and the VTA was seen by means of tyrosine hydroxylase immunohistochemistry, and tissue levels of dopamine and 3,4 dihydroxyphenylacetic acid in the NAc were also markedly increased in the EGb 761 treated animals. However, the norepinephrine tissue levels in the PVN and the NAc in the EGb 761-treated group were not significantly different from those in the controls. Together, these results suggest that administration of EGb 761 increases dopaminergic activity in the PVN and the mesolimbic system to facilitate NCE in male rats. PMID- 21640799 TI - Neurotoxicity induced by dexamethasone in the human neuroblastoma SH-SY5Y cell line can be prevented by folic acid. AB - Folic acid (folate) is a vitamin of the B-complex group that is essential for cell replication. Folate is a major determinant of one-carbon metabolism, in which S-adenosylmethionine donates methyl groups that are crucial for neurological function. Many roles for folic acid have been reported, including neuroprotective and antidepressant properties. On the other hand, increased concentrations of corticoids have proven neurotoxic effects and hypersecretion of glucocorticoids has been linked to different mood disorders. The purpose of this study was to investigate the potential protective effect of folic acid on dexamethasone-induced cellular death in SH-SY5Y neuroblastoma cell line and the possible intracellular signaling pathway involved in such effect. Exposure to 1 mM dexamethasone for 48 h caused a significant reduction of cell viability measured as 3-[4,5 dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) reduction. Exposure of SH-SY5Y cells for 72 h to increasing concentrations of folate (1-300 MUM) was not cytotoxic. However, pretreatment with folate (10-300 MUM) reduced dexamethasone-induced toxicity in a significant manner. To explore the putative intracellular signaling pathways implicated in the protective effect of folate we used different protein kinase inhibitors. The protective effect of folic acid on dexamethasone-induced neurotoxicity was reversed by the phosphatidylinositol-3 kinase/Akt (PI3K/Akt, LY294002), Ca2+/Calmodulin-dependent protein kinase II (CaMKII, KN-93), and protein kinase A (PKA, H-89) inhibitors, but not the mitogen-activated protein/extracellular signal-regulated kinase (MEK1/2, PD98059) and protein kinase C (PKC, chelerythrine) inhibitors. In conclusion, the results of this study show that folic acid can protect against dexamethasone-induced neurotoxicity and its protective mechanism is related to a signaling pathway that involves PI3K/Akt, CaMKII, and PKA. PMID- 21640800 TI - An inhibitor of eIF2 activity in the sRNA pool of eukaryotic cells. AB - Eukaryotic protein synthesis is a multi-step and highly controlled process that includes an early initiation complex containing eukaryotic initiation factor 2 (eIF2), GTP, and methionine-charged initiator methionyl-tRNA (met-tRNAi). During studies to reconstruct formation of the ternary complex containing these molecules, we detected a potent inhibitor in low molecular mass RNA (sRNA) preparations of eukaryotic tRNA. The ternary complex inhibitor (TCI) was retained in the total sRNA pool after met-tRNAi was charged by aminoacyl tRNA synthetase, co-eluted with sRNA by size exclusion chromatography, but resolved from met-tRNAi by ion exchange chromatography. The adverse effect of TCI was not overcome by high GTP or magnesium omission and was independent of GTP regeneration. Rather, TCI suppressed the rate of ternary complex formation, and disrupted protein synthesis and the accumulation of heavy polymeric ribosomes in reticulocyte lysates in vitro. Lastly, a component or components in ribosome depleted cell lysate significantly reversed TCI activity. Since assembly of the met tRNAi/eIF2/GTP ternary complex is integral to protein synthesis, awareness of TCI is important to avoid confusion in studies of translation initiation. A clear definition of TCI may also allow a better appreciation of physiologic or pathologic situations, factors, and events that control protein synthesis in vivo. PMID- 21640801 TI - Neuroblastoma cell proliferation is sensitive to changes in levels of RUNX1 and RUNX3 protein. AB - The Runt domain transcription factors Runx1 and Runx3 play crucial roles in dorsal root ganglion neurogenesis. Here we report that RUNX1 protein levels are maintained within a narrow range, despite highly variable levels of RUNX1 mRNA in neuroblastoma cell lines. Forced expression of the RUNX1 isoform AML1a (RUNX1/p27), a transcriptionally inactive competitive inhibitor, induces massive cell death, indicating that the function of RUNX1 is essential for neuroblastoma cell proliferation. Unexpectedly, over-expression of RUNX1 also induces cell death, as well as cell cycle arrest or differentiation. Furthermore, most neuroblastoma cell lines do not synthesize RUNX3 protein at detectable levels. Like RUNX1, exogenous expression also causes cell death, cell cycle arrest or differentiation. These results suggest that protein dosage of RUNX1 is critical for neuroblastoma cell growth and support a role for RUNX3 as a tumor suppressor in neuroblastoma. PMID- 21640803 TI - Lipoxygenase in Caragana jubata responds to low temperature, abscisic acid, methyl jasmonate and salicylic acid. AB - Lipoxygenase (LOX) catalyses oxygenation of free polyunsaturated fatty acids into oxylipins, and is a critical enzyme of the jasmonate signaling pathway. LOX has been shown to be associated with biotic and abiotic stress responses in diverse plant species, though limited data is available with respect to low temperature and the associated cues. Using rapid amplification of cDNA ends, a full-length cDNA (CjLOX) encoding lipoxygenase was cloned from apical buds of Caragana jubata, a temperate plant species that grows under extreme cold. The cDNA obtained was 2952bp long consisting of an open reading frame of 2610bp encoding 869 amino acids protein. Multiple alignment of the deduced amino acid sequence with those of other plants demonstrated putative LH2/ PLAT domain, lipoxygenase iron binding catalytic domain and lipoxygenase_2 signature sequences. CjLOX exhibited up- and down-regulation of gene expression pattern in response to low temperature (LT), abscisic acid (ABA), methyl jasmonate (MJ) and salicylic acid (SA). Among all the treatments, a strong up-regulation was observed in response to MJ. Data suggests an important role of jasmonate signaling pathway in response to LT in C. jubata. PMID- 21640802 TI - Molecular cloning and identification of the laspartomycin biosynthetic gene cluster from Streptomyces viridochromogenes. AB - The biosynthetic gene cluster for laspartomycins, a family of 11 amino acid peptide antibiotics, has been cloned and sequenced from Streptomyces viridochromogenes ATCC 29814. Annotation of a segment of 88912bp of S. viridochromogenes genomic sequence revealed the putative lpm cluster and its flanking regions which harbor 43 open reading frames. The lpm cluster, which spans approximately 60 kb, consists of 21 open reading frames. Those include four NRPS genes (lpmA/orf18, lpmB/orf25, lpmC/orf26 and lpmD/orf27), four genes (orfs 21, 22, 24 and 29) involved in the lipid tail biosynthesis and attachment, four regulatory genes (orfs 13, 19, 32 and 33) and three putative exporters or self resistance genes (orfs 14, 20 and 30). In addition, the gene involved in the biosynthesis of the nonproteinogenic amino acid Pip was also identified in the lpm cluster while the genes necessary for the biosynthesis of the rare residue diaminopropionic acid (Dap) were found to reside elsewhere on the chromosome. Interestingly, the dabA, dabB and dabC genes predicted to code for the biosynthesis of the unusual amino acid diaminobutyric acid (Dab) are organized into the lpm cluster even though the Dab residue was not found in the laspartomycins. Disruption of the NRPS lpmC gene completely abolished laspartomycin production in the corresponding mutant strain. These findings will allow molecular engineering and combinatorial biosynthesis approaches to expand the structural diversity of the amphomycin-group peptide antibiotics including the laspartomycins and friulimicins. PMID- 21640804 TI - Drought, salt and wounding stress induce the expression of the plasma membrane intrinsic protein 1 gene in poplar (Populus alba*P. tremula var. glandulosa). AB - Water uptake across cell membranes is a principal requirement for plant growth at both the cellular and whole-plant levels; water movement through plant membranes is regulated by aquaporins (AQPs) or major intrinsic proteins (MIPs). We examined the expression characteristics of the poplar plasma membrane intrinsic protein 1 gene (PatPIP1), a type of MIP, which was isolated from a suspension cell cDNA library of Populus alba*P. tremula var. glandulosa. Examination of protoplasts expressing the p35S-PatPIP1::sGFP fusion protein revealed that the protein was localized in the plasma membrane. Northern blot analysis revealed that the gene was strongly expressed in poplar roots and leaves. Gene expression was inducible by abiotic factors including drought, salinity, cold temperatures and wounding, and also by plant hormones including gibberellic acid, jasmonic acid and salicylic acid. Since we found that the PatPIP1 gene was strongly expressed in response to mannitol, NaCl, jasmonic acid and wounding, we propose that PatPIP1 plays an essential role in the defense of plants against water stress. PMID- 21640805 TI - Evaluation of time extrapolation factors based on the database RepDose. AB - In chemical risk assessment for many substances only short-term animal studies are available for the evaluation of long-term human exposure. Therefore usually extrapolation factors (EF) are used to extrapolate NOAELs from existing short term studies to NOAELs for long term exposure. In this report time EFs are derived, based on NOEL/C or LOEL/C ratios (short term N(L)OEL/long term N(L)OEL) from the large datasets of the database RepDose (www.fraunhofer-repdose.de) on repeated dose toxicity for oral or inhalation administration. Within a tiered approach several sources of variability, e.g. use of LOEL/C ratios or differences in dose spacing were analyzed and if needed subsequently excluded. The reduction of data variability resulted in "final" EFs datasets, which are as far as possible based on compound-specific, time-dependent differences in toxicity. For distribution functions of oral repeated dose toxicity studies characterised by GM, GSD and 90th percentiles the following data are obtained: subacute-to subchronic - GM 1.3, GSD 2.4, 90th 4.0, subacute-to-chronic - GM 3.4, GSD 3.7, 90th 18.2, and subchronic-to-chronic - GM 1.4, GSD 2.1, 90th 3.6. The number of data for inhalation exposure is limited, but with regard to systemic toxicity the derived EFs confirm the respective oral EFs. PMID- 21640806 TI - The mechanisms of drug release in poly(lactic-co-glycolic acid)-based drug delivery systems--a review. AB - Poly(D,L-lactic-co-glycolic acid) (PLGA) is the most frequently used biodegradable polymer in the controlled release of encapsulated drugs. Understanding the release mechanisms, as well as which factors that affect drug release, is important in order to be able to modify drug release. Drug release from PLGA-based drug delivery systems is however complex. This review focuses on release mechanisms, and provides a survey and analysis of the processes determining the release rate, which may be helpful in elucidating this complex picture. The term release mechanism and the various techniques that have been used to study release mechanisms are discussed. The physico-chemical processes that influence the rate of drug release and the various mechanisms of drug release that have been reported in the literature are analyzed in this review, and practical examples are given. The complexity of drug release from PLGA-based drug delivery systems can make the generalization of results and predictions of drug release difficult. However, this complexity also provides many possible ways of solving problems and modifying drug release. Basic, generally applicable and mechanistic research provides pieces of the puzzle, which is useful in the development of controlled-release pharmaceuticals. PMID- 21640807 TI - Delivery devices for the administration of paediatric formulations: overview of current practice, challenges and recent developments. AB - The European Paediatric Formulation Initiative (EuPFI), a group consisting of paediatric formulation experts from industry, academia and clinical pharmacy was founded with the aim of raising awareness of paediatric formulation issues. It is imperative that paediatric medicines can be administered accurately to ensure the correct dose is provided and that the administration device is easy to use and acceptable from the patient's and carer's perspectives. This reflection paper provides an overview of currently available paediatric administration devices and highlights some of the challenges associated with, recommendations and recent developments in delivery devices for the oral, inhaled, parenteral, nasal and ocular administration of paediatric formulations, on behalf of the EuPFI. PMID- 21640808 TI - Takeru Higuchi, the man and the scientist. AB - Madison (United States), December 7th 1941. It is early in the morning. Takeru Higuchi is listening to the radio when the programme is suddenly interrupted: part of the American naval fleet has just been destroyed in an air-raid on Pearl Harbour. For the United States, their involvement in the World War is now inevitable. For Takeru Higuchi, the son of Japanese immigrants, this was the beginning of an inner struggle. As a student of Physical Chemistry at the University of Wisconsin, he had received a fellowship from the American government; the war between the two countries pulled him in opposing directions. His austere childhood had left him with an extremely determined character and Takeru Higuchi chose to overcome his suffering by a personal dichotomy: he gave his heart to Japan but put his trust in the United States and devoted his energy to the service of science. This choice made him a man and a scientist of exceptional qualities, producing scientific work astonishing by its originality and quantity, making a major contribution to the pharmaceutical profession, particularly in the field of research. PMID- 21640809 TI - Nanostructured lipid carrier versus solid lipid nanoparticles of simvastatin: comparative analysis of characteristics, pharmacokinetics and tissue uptake. AB - Nanostructured lipid carrier (NLC) system of simvastatin was investigated for improvement in release, pharmacokinetics and biodistribution over its solid lipid nanoparticles (SLN). The NLC formulations prepared by solvent injection technique were optimized by 2(3) full factorial design. Optimized NLC was deduced on the basis of dependent variables that were analyzed using Design expert 8.0.2 software (Stat Ease, Inc., USA). Pareto charts and response surface plots were utilized to study the effect of variables on the response parameters. The optimized NLC was a suspension of nanosized homogeneous particles with significantly higher entrapment efficiency (>90%) and lower recrystallization properties (p<0.01) than SLNs. The pharmacokinetic parameters of Tc(99) labeled optimized NLC in mice, obtained using Quickcal software (Plexus, India) revealed 4.8 folds increase in bioavailability as compared to simvastatin suspension and 2.29 folds as compared to SLNs. Biodistribution study revealed preferential accumulation of NLC in the liver and this is advantageous because liver is the target organ for simvastatin. IVIVC studies demonstrated level A correlation between in vitro release and percent drug absorbed. This investigation demonstrated the superiority of NLC over SLN for improved oral delivery and it was deduced that the liquid lipid, oleic acid was the principal formulation factor responsible for the improvement in characteristics, pharmacokinetics and biodistribution of NLCs. PMID- 21640810 TI - Herbal drug patenting in India: IP potential. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Herbal drugs are gaining worldwide prominence due to their distinct advantages. Developing countries have started exploring the ethnopharmacological approach of drug discovery and have begun to file patents on herbal drugs. The expansion of R&D in Indian herbal research organizations and presence of manufacturing units at non-Indian sites is an indication of the capability to develop new products and processes. The present study attempts to identify innovations in the Indian herbal drug sector by analyzing the patenting trends in India, US and EU. METHODOLOGY: Based on key word and IPC based search at the IPO, USPTO, Esp@cenet and WIPO databases, patent applications and grant in herbal drugs by Indian applicants/assignees was collected for the last ten years (from 1st January 2001 to 31st October 2010). From this collection patents related to human therapeutic use only were selected. Analysis was performed to identify filing trends, major applicants/assignees, disease area and major plant species used for various treatments. RESULT: There is a gradual increase in patent filing through the years. In India, individual inventors have maximum applications and grants. CSIR, among research organizations and Hindustan Unilever, Avesthagen, Piramal Life Science, Sahajanand Biotech and Indus Biotech among the companies have the maximum granted patents in India, US and EU respectively. Diabetes, cancer and inflammatory disorders are the major areas for patenting in India and abroad. Recent patents are on new herbal formulations for treatment of AIDS, hepatitis, skin disorders and gastrointestinal disorders. CONCLUSION: A majority of the herbal patents applications and grants in India are with individual inventors. Claim analysis indicates that these patents include novel multi-herb compositions with synergistic action. Indian research organizations are more active than companies in filing for patents. CSIR has maximum numbers of applications not only in India but also in the US and EU. Patents by research organizations and herbal companies are on development of new processes for active compound isolation and standardization of such components in addition to new compositions for therapeutic use. Pharmaceutical companies such as Ranbaxy, Lupin and Panacea Biotec are increasingly patenting on herbal drugs. There is increased patenting activity related to diabetes, cancer, cardiovascular diseases, asthma and arthritis in India and abroad. PMID- 21640811 TI - Long-lasting effects of perinatal asphyxia on exploration, memory and incentive downshift. AB - Perinatal asphyxia remains as one of the most important causes of death and disability in children, without an effective treatment. Moreover, little is known about the long-lasting behavioral consequences of asphyxia at birth. Therefore, the main aim of the present study was to investigate the motor, emotional and cognitive functions of adult asphyctic rats. Experimental subjects consisted of rats born vaginally (CTL), by cesarean section (C+), or by cesarean section following 19 min of asphyxia (PA). At three months of age, animals were examined in a behavioral test battery including elevated plus maze, open field, Morris water maze, and an incentive downshift procedure. Results indicated that groups did not differ in anxiety-related behaviors, although a large variability was observed in the asphyctic group and therefore, the results are not completely conclusive. In addition, PA and C+ rats showed a deficit in exploration of new environments, but to a much lesser extent in the latter group. Spatial reference and working memory impairments were also found in PA rats. Finally, when animals were downshifted from a 32% to a 4% sucrose solution, an attenuated suppression of consummatory behavior was observed in PA rats. These results confirmed and extended those reported previously about the behavioral alterations associated with acute asphyxia around birth. PMID- 21640812 TI - Yersinia pestis TIR-domain protein forms dimers that interact with the human adaptor protein MyD88. AB - Recent research has highlighted the presence of Toll/Interleukin 1 receptor (TIR) domain proteins (Tdps) in a range of bacteria, suggested to form interactions with the human adaptor protein MyD88 and inhibit intracellular signaling from Toll-like receptors (TLRs). A Tdp has been identified in Yersinia pestis (YpTdp), a highly pathogenic bacterium responsible for plague. Expression of a number of YpTIR constructs of differing lengths (YpTIR1, S130-A285; YpTIR2, I137-I273; YpTIR3, I137-246; YpTIR4, D107-S281) as fusions with an N-terminal GB1 tag (the B1 immunoglobulin domain of Streptococcal protein G) yielded high levels of soluble protein. Subsequent purification yielded 4-6 mg/L pure, folded protein. Thrombin cleavage allowed separation of the GB1 tag from YpTIR4 resulting in folded protein after cleavage. Nuclear magnetic resonance spectroscopy, size exclusion chromatography, SDS-PAGE analysis and static light scattering all indicate that the YpTIR forms dimers. Generation of a double Cys-less mutant resulted in an unstable protein containing mainly monomers indicating the importance of disulphide bonds in dimer formation. In addition, the YpTIR constructs have been shown to interact with the human adaptor protein MyD88 using 2D NMR and GST pull down. YpTIR is an excellent candidate for further study of the mechanism of action of pathogenic bacterial Tdps. PMID- 21640813 TI - Cytotoxicity of Triptolide and Triptolide loaded polymeric micelles in vitro. AB - Triptolide (TP), a diterpenoid triepoxide purified from the Chinese herb Tripterygium wilfordii Hook F is characterized by strong anti-tumor effects on various cancer cells. Except its anti-tumor effects, TP also shows multiple pharmacological side activities, such as anti-inflammatory, immune-suppressive and male anti-fertility. In order to reduce these side effects, especially the immuno-suppressive activity when used to cure cancer, a novel polymeric micelle system containing TP (TP-PM) was constructed. The immune-modulation effects of TP PM have been evaluated by previous studies. In this study, we compared the cytotoxicity of TP and TP-PM on Jurkat and HT29 cells. Therefore, we determined the cell viability, membrane integrity, cell proliferation, apoptosis, and caspase 3/7 activity after exposure to TP and TP-PM. The results demonstrated that actually low concentrated TP and TP-PM could induce an inhibition of cell growth and proliferation as well as membrane damage in both tumor cell lines. TP and TP-PM induced apoptosis and caused activation of caspase 3/7 even at low concentrations. Both formulations destroyed the membrane of Jurkat cells, nevertheless, TP-PM showed stronger pernicious effects. In general, TP-PM induced in both tested cell lines stronger effects than TP. Therefore, polymeric micelles can be considered as promising carriers for TP in cancer therapy. PMID- 21640814 TI - Decoding the genome beyond sequencing: the new phase of genomic research. AB - While our understanding of gene-based biology has greatly improved, it is clear that the function of the genome and most diseases cannot be fully explained by genes and other regulatory elements. Genes and the genome represent distinct levels of genetic organization with their own coding systems; Genes code parts like protein and RNA, but the genome codes the structure of genetic networks, which are defined by the whole set of genes, chromosomes and their topological interactions within a cell. Accordingly, the genetic code of DNA offers limited understanding of genome functions. In this perspective, we introduce the genome theory which calls for the departure of gene-centric genomic research. To make this transition for the next phase of genomic research, it is essential to acknowledge the importance of new genome-based biological concepts and to establish new technology platforms to decode the genome beyond sequencing. PMID- 21640815 TI - Genome-wide identification of novel microRNAs and their target genes in the human parasite Schistosoma mansoni. AB - Mature microRNAs (miRNAs) are small, non-coding regulatory RNAs which can elicit post-transcriptional repression of mRNA levels of target genes. Here, we report the identification of 67 mature and 42 precursor miRNAs in the Schistosoma mansoni parasite. The evolutionarily conserved S. mansoni miRNAs consisted of 26 precursor miRNAs and 35 mature miRNAs, while we identified 16 precursor miRNAs and 32 mature miRNAs that displayed no conservation. These S. mansoni miRNAs are located on seven autosomal chromosomes and a sex (W) chromosome. miRNA expansion through gene duplication was suggested for at least two miRNA families miR-71 and mir-2. miRNA target finding analysis identified 389 predicted mRNA targets for the identified miRNAs and suggests that the sma-mir-71 may be involved in female sexual maturation. Given the important roles of miRNAs in animals, the identification and characterization of miRNAs in S. mansoni will facilitate novel approaches towards prevention and treatment of Schistosomiasis. PMID- 21640816 TI - The social environment and IL-6 in rats and humans. AB - Inflammatory cytokine levels predict a wide range of human diseases including depression, cardiovascular disease, type 2 diabetes, autoimmune disease, general morbidity, and mortality. Stress and social experiences throughout the lifecourse have been associated with inflammatory processes. We conducted studies in humans and laboratory rats to examine the effect of early life experience and adult social position in predicting IL-6 levels. Human participants reported family homeownership during their childhood and current subjective social status. Interleukin-6 (IL-6) was measured from oral mucosal transudate. Rats were housed in groups of three, matched for quality of maternal care received. Social status was assessed via competition for resources, and plasma IL-6 was assessed in adulthood. In both humans and rats, we identified an interaction effect; early social experience moderated the effect of adult social status on IL-6 levels. Rats that experienced low levels of maternal care and people with low childhood socioeconomic status represented both the highest and lowest levels of IL-6 in adulthood, depending on their social status as young adults. The predicted interaction held for non-Hispanic people, but did not occur among Hispanic individuals. Adversity early in life may not have a monotonically negative effect on adult health, but may alter biological sensitivity to later social experiences. PMID- 21640817 TI - Early and adult hippocampal TGF-beta1 overexpression have opposite effects on behavior. AB - TGF-beta1 is an anti-inflammatory cytokine that is augmented in the brain of autistic patients and that can affect brain development. In this work, we studied the effects of overexpressing TGF-beta1 in the dentate gyrus of adult or young mice on behavior. TGF-beta1 overexpression during postnatal development led to a long-term decrease in social interaction and to long-term increases in self grooming and depression-related behaviors. Our analysis shows that these behavioral changes correlate with the long-term downregulation of TGF-beta1 and IL-6 expression in the dentate gyrus, as well as to decreases in the mRNA levels of the synaptic protein neuroligin 3 and in the number of Reelin-positive neurons in the dentate gyrus. In contrast, chronic expression of TGF-beta1 during adulthood led to transient opposite effects on these behaviors. These results show a central role of hippocampal TGF-beta1 in the programming and modulation of social interaction, repetitive behavior and depression-related behavior. Finally, our data suggest a role of hippocampal TGF-beta1 and early-life neuroinflammation in the development of the behavioral alterations observed in autism spectrum disorders. PMID- 21640818 TI - Lysosomal thiol reductase negatively regulates autophagy by altering glutathione synthesis and oxidation. AB - Redox regulation is critical for a number of cellular functions and has been implicated in the etiology and progression of several diseases, such as cardiovascular diseases, neurodegenerative diseases, and cancer. It has been shown that, in the absence of gamma-interferon inducible lysosomal thiol reductase (GILT), cells are under increased oxidative stress with higher superoxide levels and decreased stability, expression, and function of mitochondrial manganese superoxide dismutase (SOD2). Here, we further elucidate the role of GILT in the homeostatic regulation of oxidative stress. We show that GILT-deficient fibroblasts exhibit reduced glutathione levels, shift in GSSG/GSH ratio toward the oxidized form, and accumulate dysfunctional mitochondria. Redox sensitive pathways involving Erk1/2 activation and nuclear high mobility group box 1 (HMGB1) protein cytosolic translocation are both activated and associated with increased autophagy in GILT-/- fibroblasts. We hypothesize that these events are responsible for degrading the damaged mitochondria and mitochondrial SOD2 in the absence of GILT. This is the first time to our knowledge that a lysosomal enzyme has been implicated in global effects within the cell. PMID- 21640819 TI - A tryparedoxin-dependent peroxidase protects African trypanosomes from membrane damage. AB - Hydroperoxide detoxification in African trypanosomes is achieved by 2-Cys peroxiredoxin (TXNPx)- and non-selenium glutathione peroxidase (Px)-type enzymes which both obtain their reducing equivalents from the unique trypanothione/tryparedoxin system. Previous RNA interference approaches revealed that the cytosolic TXNPx and the Px-type enzymes are essential for Trypanosoma brucei. Because of partially overlapping in vitro substrate specificities and subcellular localisation the physiological function of the individual enzymes was not yet clear. As shown here, TXNPx and Px are expressed at comparable levels and in their active reduced state. Px-overexpressing parasites were less sensitive toward linoleic acid hydroperoxide but not hydrogen peroxide. Kinetic studies confirmed that Px-but not TXNPx-reduces lipophilic hydroperoxides including phospholipids with high efficiency. Most interestingly, the severe proliferation defect of Px-depleted bloodstream cells could be rescued by Trolox, but not by hydrophilic antioxidants, in the medium. This allowed us to knock-out the three Px genes individually and thus to distinguish their in vivo role. Deletion of the cytosolic Px I and II resulted in extremely fast membrane peroxidation followed by cell lysis. Cells lacking specifically the mitochondrial Px III showed a transient growth retardation and cardiolipin peroxidation but adapted within 24h to normal proliferation. PMID- 21640821 TI - Polycystic kidney disease: the complexity of planar cell polarity and signaling during tissue regeneration and cyst formation. AB - Autosomal Dominant Polycystic Kidney Disease (ADPKD) is an inherited systemic disease with intrarenal cystogenesis as its primary characteristic. A variety of mouse models provided information on the requirement of loss of balanced polycystin levels for initiation of cyst formation, the role of proliferation in cystogenesis and the signaling pathways involved in cyst growth and expansion. Here we will review the involvement of different signaling pathways during renal development, renal epithelial regeneration and cyst formation in ADPKD, focusing on planar cell polarity (PCP) and oriented cell division (OCD). This will be discussed in context of the hypothesis that aberrant PCP signaling causes cyst formation. In addition, the role of the Hippo pathway, which was recently found to be involved in cyst growth and tissue regeneration, and well-known for regulating organ size control, will be reviewed. The fact that Hippo signaling is linked to PCP signaling makes the Hippo pathway a novel cascade in cystogenesis. The newly gained understanding of the complex signaling network involved in cystogenesis and disease progression, not only necessitates refining of the current hypothesis regarding initiation of cystogenesis, but also has implications for therapeutic intervention strategies. This article is part of a Special Issue entitled: Polycystic Kidney Disease. PMID- 21640820 TI - Low-level lead exposure triggers neuronal apoptosis in the developing mouse brain. AB - While the toxic effects of lead have been recognized for millennia, it has remained a significant public health concern due to its continued use and toxicological potential. Of particular interest is the increased susceptibility of young children to the toxic effects of lead. Although the exact mechanism(s) for lead toxicity is currently not well understood, research has established that it can be a potent NMDA antagonist. Previous research has established that exposure to NMDA antagonists during the brain growth spurt period (first 2 weeks of life in mice) can produce apoptotic neurodegeneration throughout the brain. Based on this information, the ability of lead exposure (two injections of 350 mg/kg lead 4h apart) to produce apoptosis in the neonatal mouse brain was assessed histologically 8-24h after treatment using activated caspase-3 immunohistochemistry, De Olmos silver technique, Nissl staining, and electron microscopy. Lead exposure produced significant neurodegeneration in the caudate/putamen, hippocampus, subiculum, and superficial and deep cortical layers of the frontal cortical regions. Further ultrastructural examination revealed cellular profiles consistent with apoptotic cell death. Statistical results showed that lead exposure significantly increased apoptotic neurodegeneration above that seen in normal controls in animals treated at postnatal day 7, but not on day 14. The results of this study may provide a basis for further elucidation of mechanisms through which the immature nervous system may be particularly susceptible to lead exposure. PMID- 21640822 TI - Differential resistance of insect sodium channels with kdr mutations to deltamethrin, permethrin and DDT. AB - Knockdown resistance (kdr) in insects, caused by inherited nucleotide polymorphisms in the voltage-gated sodium channel (VGSC) gene, is a major threat to the efficacy of pyrethroid insecticides. Classic kdr, resulting from an L1014F substitution in the VGSC is now present in numerous pest species. Two other substitutions at the L1014 locus have also been reported, L1014S and L1014H. Here we have used expression of L1014 modified Drosophila para VGSCs in Xenopus oocytes with two-electrode voltage clamp to characterise all three mutations. The mutations L1014F and L1014H caused significant depolarizing shifts in the half activation voltage (V(50,act)) from -17.3 mV (wild-type) to -13.1 and -13.5 mV respectively, whereas L1014S caused no shift in V(50,act) but its currents decayed significantly faster than wild-type channels. Treatment of the wild-type channel with deltamethrin (>= 1 nM), permethrin (>= 30 nM) or DDT (>= 1 MUM) resulted in hyperpolarizing shifts in V(50,act). Deltamethrin, permethrin and DDT also produced "tail currents" with EC50s of 0.043, 0.40 and 65 MUM and maximum modifications of 837, 325 and 7% respectively. L1014F provided a high level of resistance against all insecticides for both measured parameters. L1014H most effectively combated deltamethrin induced tail currents while L1014S strongly resisted the large DDT induced shifts in V(50,act). We conclude that L1014H and L1014S may have arisen through heavy exposure to specific pyrethroids and DDT respectively. PMID- 21640823 TI - Complete sequences of mitochondria genomes of Aedes aegypti and Culex quinquefasciatus and comparative analysis of mitochondrial DNA fragments inserted in the nuclear genomes. AB - We present complete sequences of the mitochondrial genomes for two important mosquitoes, Aedes aegypti and Culex quinquefasciatus, that are major vectors of dengue virus and lymphatic filariasis, respectively. The A. aegypti mitochondrial genome is 16,655 bp in length and that of C. quinquefasciatus is 15,587 bp, yet both contain 13 protein coding genes, 22 transfer RNA (tRNA) genes, one 12S ribosomal RNA (rRNA) gene, one 16S rRNA gene and a control region (CR) in the same order. The difference in the genome size is due to the difference in the length of the control region. We also analyzed insertions of nuclear copies of mtDNA-like sequences (NUMTs) in a comparative manner between the two mosquitoes. The NUMT sequences occupy ~0.008% of the A. aegypti genome and ~0.001% of the C. quinquefasciatus genome. Several NUMTs were found localized in the introns of predicted protein coding genes in both genomes (32 genes in A. aegypti but only four in C. quinquefasciatus). None of these NUMT-containing genes had an ortholog between the two species or had paralogous copies within a genome that was also NUMT-containing. It was further observed that the NUMT-containing genes were relatively longer but had lower GC content compared to the NUMT-less paralogous copies. Moreover, stretches of homologies are present among the genic and non genic NUMTs that may play important roles in genomic rearrangement of NUMTs in these genomes. Our study provides new insights on understanding the roles of nuclear mtDNA sequences in genome complexities of these mosquitoes. PMID- 21640825 TI - On the intrinsic inevitability of cancer: from foetal to fatal attraction. AB - The cracks in the paradigm of oncogenic mutations and somatic evolution as driving force of tumorigenesis, lucidly exposed by the dynamic heterogeneity of "cancer stem cells" or the diffuse results of cancer genome sequencing projects, indicate the need for a more encompassing theory of cancer that reaches beyond the current proximate explanations based on individual genetic pathways. One such integrative concept, derived from first principles of the dynamics of gene regulatory networks, is that cancerous cell states are attractor states, just like normal cell types are. Here we extend the concept of cancer attractors to illuminate a more profound property of cancer initiation: its inherent inevitability in the light of metazoan evolution. Using Waddington's Epigenetic Landscape as a conceptual aid, for which we present a mathematical and evolutionary foundation, we propose that cancer is intrinsically linked to ontogenesis and phylogenesis. This explanatory rather than enumerating review uses a formal argumentation structure that is atypical in modern experimental biology but may hopefully offer a new coherent perspective to reconcile many conflicts between new findings and the old thinking in the categories of linear oncogenic pathways. PMID- 21640824 TI - Alterations in the cholinergic system after frontal cortical infarction in rat brain: pharmacological magnetic resonance imaging of muscarinic receptor responsiveness and stereological analysis of cholinergic forebrain neurons. AB - Vascular cognitive impairment has been related to dysfunction of the central cholinergic system. Studies exploring the putative relationship between vascular cognitive impairment and cholinergic dysfunction have largely been aimed at symptomatic cholinergic treatment rather than focusing on etiological and pathological factors. The present study characterizes chronic responses of the cholinergic system to focal cerebral infarction. Two separate experiments investigated changes in receptor responsiveness versus changes in cell number after photothrombotic infarction of the frontal cortex in rat brain. First, we conducted pharmacological magnetic resonance imaging (phMRI) together with pilocarpine injection to assess relative cerebral blood volume (CBV) responses related to cholinergic muscarinic receptor activation. PhMRI was conducted at 1 and 3 weeks after photothrombotic infarction of either the left or right frontal cortex. Second, stereological assessment was performed on choline acetyltransferase (ChAT)-immunostained sections to determine cholinergic cell body count in several basal forebrain nuclei at 4 weeks after infarction. Significant reductions in relative CBV responses were observed both inside the ischemic area at 1 and 3 weeks, and in areas distant from the lesion at 3 weeks after right-sided frontal cortical infarction. In contrast, cholinergic cell number remained unchanged. These results demonstrate that cholinergic receptor responsiveness may be significantly altered following cerebral infarction, while projecting cholinergic cells are preserved. PMID- 21640826 TI - Biotechnology techniques for the development of new tumor specific peptides. AB - Peptides, proteins and antibodies are promising candidates as carriers for radionuclides in endoradiotherapy. This novel class of pharmaceuticals offers a great potential for the targeted therapy of cancer. The fact that some receptors are overexpressed in several tumor types and can be targeted by small peptides, proteins or antibodies conjugated to radionuclides has been used in the past for the development of peptide endoradiotherapeutic agents such as (90)Y-DOTATOC or radioimmunotherapy of lymphomas with Zevalin. These procedures have been shown to be powerful options for the treatment of cancer patients. Design of new peptide libraries and scaffolds combined with biopanning techniques like phage and ribosome display may lead to the discovery of new specific ligands for target structures overexpressed in malignant tumors. Display methods are high throughput systems which select for high affinity binders. These methods allow the screening of a vast amount of potential binding motifs which may be exposed to either cells overexpressing the target structures or in a cell-free system to the protein itself. Labelling these binders with radionuclides creates new potential tracers for application in diagnosis and endoradiotherapy. This review highlights the advantages and problems of phage and ribosome display for the identification and evaluation of new tumor specific peptides. PMID- 21640827 TI - Effects of L-arginine on solubilization and purification of plant membrane proteins. AB - Biochemical analysis of membrane proteins is problematic at the level of solubilization and/or purification because of their hydrophobic nature. Here, we developed methods for efficient solubilization and purification of membrane proteins using L-arginine. The addition of 100 mM of basic amino acids (L arginine, L-lysine, and L-ornithine) to a detergent-containing solubilization buffer enhanced solubilization (by 2.6-4.3 fold) of a model membrane protein polygalacturonic acid synthase. Of all the amino acids, arginine was the most effective additive for solubilization of this membrane protein. Arginine addition also resulted in the best solubilization of other plant membrane proteins. Next, we examined the effects of arginine on purification of a model membrane protein. In anion-exchange chromatography, the addition of arginine to the loading and elution buffers resulted in a greater recovery of a membrane protein. In ultrafiltration, the addition of arginine to a protein solution significantly improved the recovery of a membrane protein. These results were thought to be due to the properties of arginine that prevent aggregation of hydrophobic proteins. Taken together, the results of our study showed that arginine is useful for solubilization and purification of aggregate-prone membrane proteins. PMID- 21640829 TI - Production and crystallization of processing alpha-glucosidase I: Pichia pastoris expression and a two-step purification toward structural determination. AB - Eukaryotic N-glycoprotein processing in the endoplasmic reticulum begins with the catalytic action of processing alpha-glucosidase I (alphaGlu). alphaGlu trims the terminal glucose from nascent glycoproteins in an inverting-mechanism glycoside hydrolysis reaction. alphaGlu has been studied in terms of kinetic parameters and potential key residues; however, the active site is unknown. A structural model would yield important insights into the reaction mechanism. A model would also be useful in developing specific therapeutics, as alphaGlu is a viable drug target against viruses with glycosylated envelope proteins. However, due to lack of a high-yielding overexpression and purification scheme, no eukaryotic structural model of alphaGlu has been determined. To address this issue, we overexpressed the Saccharomyces cerevisiae soluble alphaGlu, Cwht1p, in the host Pichia pastoris. It was purified in a simple two-step protocol, with a final yield of 4.2mg Cwht1p per liter of growth culture. To test catalytic activity, we developed a modified synthesis of a tetrasaccharide substrate, Glc(3)ManOMe. Cwht1p with Glc(3)ManOMe shows a K(m) of 1.26 mM. Cwht1p crystals were grown and subjected to X-ray irradiation, giving a complete diffraction dataset to 2.04 A resolution. Work is ongoing to obtain phases so that we may further understand this fundamental member of the N-glycosylation pathway through the discovery of its molecular structure. PMID- 21640830 TI - Generation and evaluation of mammalian secreted and membrane protein expression libraries for high-throughput target discovery. AB - Expressed protein libraries are becoming a critical tool for new target discovery in the pharmaceutical industry. In order to get the most meaningful and comprehensive results from protein library screens, it is essential to have library proteins in their native conformation with proper post-translation modifications. This goal is achieved by expressing untagged human proteins in a human cell background. We optimized the transfection and cell culture conditions to maximize protein expression in a 96-well format so that the expression levels were comparable with the levels observed in shake flasks. For detection purposes, we engineered a 'tag after stop codon' system. Depending on the expression conditions, it was possible to express either native or tagged proteins from the same expression vector set. We created a human secretion protein library of 1432 candidates and a small plasma membrane protein set of about 500 candidates. Utilizing the optimized expression conditions, we expressed and analyzed both libraries by SDS-PAGE gel electrophoresis and Western blotting. Two thirds of secreted proteins could be detected by Western-blot analyses; almost half of them were visible on Coomassie stained gels. In this paper, we describe protein expression libraries that can be easily produced in mammalian expression systems in a 96-well format, with one protein expressed per well. The libraries and methods described allow for the development of robust, high-throughput functional screens designed to assay for protein specific functions associated with a relevant disease-specific activity. PMID- 21640828 TI - Expression and purification of 15N- and 13C-isotope labeled 40-residue human Alzheimer's beta-amyloid peptide for NMR-based structural analysis. AB - Amyloid fibrils of Alzheimer's beta-amyloid peptide (Abeta) are a primary component of amyloid plaques, a hallmark of Alzheimer's disease (AD). Enormous attention has been given to the structural features and functions of Abeta in amyloid fibrils and other type of aggregates in associated with development of AD. This report describes an efficient protocol to express and purify high quality 40-residue Abeta(1-40), the most abundant Abeta in brains, for structural studies by NMR spectroscopy. Over-expression of Abeta(1-40) with glutathione S transferase (GST) tag connected by a Factor Xa recognition site (IEGR(?)) in Escherichia coli resulted in the formation of insoluble inclusion bodies even with the soluble GST tag. This problem was resolved by efficient recovery of the GST-Abeta fusion protein from the inclusion bodies using 0.5% (w/v) sodium lauroyl sarcosinate as solubilizing agent and subsequent purification by affinity chromatography using a glutathione agarose column. The removal of the GST tag by Factor Xa enzymatic cleavage and purification by HPLC yielded as much as ~7 mg and ~1.5mg of unlabeled Abeta(1-40) and uniformly (15)N- and/or (13)C-protein Abeta(1-40) from 1L of the cell culture, respectively. Mass spectroscopy of unlabeled and labeled Abeta and (1)H/(15)N HSQC solution NMR spectrum of the obtained (15)N-labeled Abeta in the monomeric form confirmed the expression of native Abeta(1-40). It was also confirmed by electron micrography and solid-state NMR analysis that the purified Abeta(1-40) self-assembles into beta-sheet rich amyloid fibrils. To the best of our knowledge, our protocol offers the highest yields among published protocols for production of recombinant Abeta(1-40) samples that are amendable for an NMR-based structural analysis. The protocol may be applied to efficient preparation of other amyloid-forming proteins and peptides that are (13)C- and (15)N-labeled for NMR experiments. PMID- 21640831 TI - Molecular recognition force spectroscopy of a specific lectin-carbohydrate interaction at single-molecule level. AB - Carbohydrates are involved in many essential biological recognition processes in physiological and pathological states. Thus, it is important to understand the mechanism of protein-carbohydrate interactions at molecular level. In the present study, molecular recognition force spectroscopy was applied to investigate the interactions between RCA120, a lectin from Ricinus communis, and galactose (Gal) and asialofetuin (ASF) at the single-molecule level. RCA120 coupled to the AFM tip could specifically recognize Gal and ASF, respectively. The unbinding forces of RCA120-Gal and RCA120-ASF increase linearly with the logarithm of loading rate. The results reveal that the binding capability of RCA120 toward Gal is weaker than that of ASF, implicating a multivalent effect in the RCA120-ASF interaction. PMID- 21640832 TI - Molecular characterization and expression analysis of six peroxiredoxin paralogous genes in gilthead sea bream (Sparus aurata): insights from fish exposed to dietary, pathogen and confinement stressors. AB - The aim of this work was to underline the physiological role of the antioxidant peroxiredoxin (PRDX) family in gilthead sea bream (Sparus aurata L.), a perciform fish extensively cultured in the Mediterranean area. First, extensive BLAST searches were done on the gilthead sea bream cDNA database of the AQUAMAX European Project (www.sigenae.org/iats), and six contigs were unequivocally identified as PRDX1-6 after sequence completion by RT-PCR. The phylogenetic analysis evidenced three major clades corresponding to PRDX1-4 (true 2-Cyst PRDX subclass), PRDX5 (atypical 2-Cys PRDX subclass) and PRDX6 (1-Cys PRDX subclass) that reflected the present hierarchy of vertebrates. However, the PRDX2 branch of modern fish including gilthead sea bream was related to the monophyletic PRDX1 node rather than to PRDX2 cluster of mammals and primitive fish, which probably denotes the acquisition of novel functions through vertebrate evolution. Transcriptional studies by means of quantitative real-time PCR evidenced a ubiquitous PRDX gene expression that was tissue specific for each PRDX isoform. In a second set of transcriptional studies, liver and head kidney were chosen as target tissues in fish challenged with i) the intestinal parasite Enteromyxum leei, ii) a plant oil (VO) diet with deficiencies in essential fatty acids and iii) prolonged exposure to high-rearing densities. These studies showed that PRDX genes were highly and mostly constitutively expressed in the liver and were not affected by dietary intervention or high density. In contrast, head kidney was highly sensitive to the different experimental challenges: significantly lower values were found for PRDX5 in the three trials, for PRDX6 in parasitized and high density fish and for PRDX1 in parasitized and VO fish. PRDX2, 3 and 5 were decreased only in VO, high density and parasitized animals, respectively. These findings would highlight the role of PRDXs as integrative and highly predictive biomarkers of health and welfare in fish and gilthead sea bream in particular. PMID- 21640833 TI - Spatiotemporal dynamics of epileptiform propagations: imaging of human brain slices. AB - Seizure activities often originate from a localized region of the cerebral cortex and spread across large areas of the brain. The properties of these spreading abnormal discharges may account for clinical phenotypes in epilepsy patients, although the manner of their propagation and the underlying mechanisms are not well understood. In the present study we performed flavoprotein fluorescence imaging of cortical brain slices surgically resected from patients with partial epilepsy caused by various symptomatic lesions. Elicited neural activities in the epileptogenic tissue spread horizontally over the cortex momentarily, but those in control tissue taken from patients with brain tumors who had no history of epilepsy demonstrated only localized responses. Characteristically, the epileptiform propagation comprised early and late phases. When the stimulus intensity was changed gradually, the early phase showed an all-or-none behavior, whereas the late phase showed a gradual increase in the response. Moreover, the two phases were propagated through different cortical layers, suggesting that they are derived from distinct neural circuits. Morphological investigation revealed the presence of hypertrophic neurons and loss of dendritic spines, which might participate in the aberrant activities observed by flavoprotein fluorescence imaging. These findings indicate that synchronized activities of the early phase may play a key role in spreading abnormal discharges in human cortical epilepsies. PMID- 21640834 TI - Neuromagnetic representation of musical register information in human auditory cortex. AB - Pulse-resonance sounds like vowels or instrumental tones contain acoustic information about the physical size of the sound source (pulse rate) and body resonators (resonance scale). Previous research has revealed correlates of these variables in humans using functional neuroimaging. Here, we report two experiments that use magnetoencephalography to study the neuromagnetic representations of pulse rate and resonance scale in human auditory cortex. In Experiment 1, auditory evoked fields were recorded from nineteen subjects presented with French horn tones, the pulse rate and resonance scale of which had been manipulated independently using a mucoder. In Experiment 2, fifteen subjects listened to French horn tones which differed in resonance scale but which lacked pulse rate cues. The resulting cortical activity was evaluated by spatio-temporal source analysis. Changes in pulse rate elicited a well-defined N1m component with cortical generators located at the border between Heschl's gyrus and planum temporale. Changes in resonance scale elicited a second, independent, N1m component located in planum temporale. Our results demonstrate that resonance scale can be distinguished in its neuromagnetic representation from cortical activity related to the sound's pulse rate. Moreover, the existence of two separate components in the N1m sensitive to register information highlights the importance of this time window for the processing of frequency information in human auditory cortex. PMID- 21640836 TI - Attention, biological motion, and action recognition. AB - Interacting with others in the environment requires that we perceive and recognize their movements and actions. Neuroimaging and neuropsychological studies have indicated that a number of brain regions, particularly the superior temporal sulcus, are involved in a number of processes essential for action recognition, including the processing of biological motion and processing the intentions of actions. We review the behavioral and neuroimaging evidence suggesting that while some aspects of action recognition might be rapid and effective, they are not necessarily automatic. Attention is particularly important when visual information about actions is degraded or ambiguous, or if competing information is present. We present evidence indicating that neural responses associated with the processing of biological motion are strongly modulated by attention. In addition, behavioral and neuroimaging evidence shows that drawing inferences from the actions of others is attentionally demanding. The role of attention in action observation has implications for everyday social interactions and workplace applications that depend on observing, understanding and interpreting actions. PMID- 21640835 TI - Discriminating schizophrenia and bipolar disorder by fusing fMRI and DTI in a multimodal CCA+ joint ICA model. AB - Diverse structural and functional brain alterations have been identified in both schizophrenia and bipolar disorder, but with variable replicability, significant overlap and often in limited number of subjects. In this paper, we aimed to clarify differences between bipolar disorder and schizophrenia by combining fMRI (collected during an auditory oddball task) and diffusion tensor imaging (DTI) data. We proposed a fusion method, "multimodal CCA+ joint ICA", which increases flexibility in statistical assumptions beyond existing approaches and can achieve higher estimation accuracy. The data collected from 164 participants (62 healthy controls, 54 schizophrenia and 48 bipolar) were extracted into "features" (contrast maps for fMRI and fractional anisotropy (FA) for DTI) and analyzed in multiple facets to investigate the group differences for each pair-wised groups and each modality. Specifically, both patient groups shared significant dysfunction in dorsolateral prefrontal cortex and thalamus, as well as reduced white matter (WM) integrity in anterior thalamic radiation and uncinate fasciculus. Schizophrenia and bipolar subjects were separated by functional differences in medial frontal and visual cortex, as well as WM tracts associated with occipital and frontal lobes. Both patients and controls showed similar spatial distributions in motor and parietal regions, but exhibited significant variations in temporal lobe. Furthermore, there were different group trends for age effects on loading parameters in motor cortex and multiple WM regions, suggesting that brain dysfunction and WM disruptions occurred in identified regions for both disorders. Most importantly, we can visualize an underlying function-structure network by evaluating the joint components with strong links between DTI and fMRI. Our findings suggest that although the two patient groups showed several distinct brain patterns from each other and healthy controls, they also shared common abnormalities in prefrontal thalamic WM integrity and in frontal brain mechanisms. PMID- 21640837 TI - Fractional anisotropy of cerebral white matter and thickness of cortical gray matter across the lifespan. AB - We examined age trajectories of fractional anisotropy (FA) of cerebral white matter (WM) and thickness of cortical gray matter (GM) in 1031 healthy human subjects (aged 11-90 years). Whole-brain FA and GM thickness values followed quadratic trajectories with age but the relationship between them was linear, indicating that a putative biological mechanism may explain the non-linearity of their age trajectories. Inclusion of the FA values into the quadratic model of the whole-brain and regional GM thickness changes with age made the effect of the age(2) term no longer significant for the whole-brain GM thickness and greatly reduced its significance for regional GM thickness measurements. The phylogenetic order of cerebral myelination helped to further explain the intersubject variability in GM thickness. FA values for the early maturing WM were significantly better (p=10(-6)) at explaining variability in GM thickness in maturing (aged 11-20) subjects than FA values for the late maturing WM. The opposite trend was observed for aging subjects (aged 40-90) where FA values for the late maturing WM were better (p=10(-16)) at explaining the variability in GM thickness. We concluded that the non-linearity of the age trajectory for GM thickness, measured from T1-weighted MRI, was partially explained by the heterogeneity and the heterochronicity of the age-related changes in the microintegrity of cerebral WM. We consider these findings as the evidence that the measurements of age-related changes in GM thickness and FA are driven, in part, by a common biological mechanism, presumed to be related to changes in cerebral myelination. PMID- 21640838 TI - Vigorous exercise increases brain lactate and Glx (glutamate+glutamine): a dynamic 1H-MRS study. AB - Vigorous exercise increases lactate and glucose uptake by the brain in excess of the increase in brain oxygen uptake. The metabolic fate of this non-oxidized carbohydrate entering the brain is poorly understood, but accumulation of lactate in the brain and/or increased net synthesis of amino acid neurotransmitters are possible explanations. Previous proton magnetic resonance spectroscopy (1H-MRS) studies using conventional pulse sequences have not detected changes in brain lactate following exercise. This contrasts with 1H-MRS studies showing increased brain lactate when blood lactate levels are raised by an intravenous infusion of sodium lactate. Using a J-editing 1H-MRS technique for measuring lactate, we demonstrated a significant 19% increase in lactate in the visual cortex following graded exercise to approximately 85% of predicted maximum heart rate. However, the magnitude of the increase was insufficient to account for more than a small fraction of the non-oxidized carbohydrate entering the brain with exercise. We also report a significant 18% increase in Glx (combined signal from glutamate and glutamine) in visual cortex following exercise, which may represent an activity dependent increase in glutamate. Future studies will be necessary to test the hypothesis that non-oxidized carbohydrate entering the brain during vigorous exercise is directed, in part, toward increased net synthesis of amino acid neurotransmitters. The possible relevance of these findings to panic disorder and major depression is discussed. PMID- 21640840 TI - Associative memory deficits in mild cognitive impairment: the role of hippocampal formation. AB - Neuropathological events featuring early stages of Alzheimer's disease (AD) appear in the entorhinal cortex (EC), subiculum (SB) and cornu ammonis 1 (CA1) of hippocampus, which may account for associative memory deficits in non-demented people with mild cognitive impairment (MCI). To test this hypothesis in vivo, we investigated whether volume changes in these regions are related to failures in associative memory in MCI as compared to cognitively normal (CN) elderly subjects. Volume changes in EC and hippocampal subfields were determined by using deformation-based morphometry techniques applied to probabilistic cytoarchitectonic maps derived from post mortem human brains. CN subjects were distinguished from MCI patients by firstly identifying local volume differences in EC and hippocampus, and then evaluating the way in which these anatomical changes correlated with performance in a non-intentional face-location association task. MCI patients not only performed worse than CN elders in building new associations, but they were further unable to benefit from semantic encoding to improve episodic binding. According to our initial hypothesis, local volume reductions in both EC and hippocampal CA accounted for group differences in associative memory whereas atrophy in CA, but not in EC, accounted for semantic encoding of associations. Two main conclusions can be drawn from the present study: i) access to semantic information during encoding does not reduce the episodic deficit in MCI; and ii) EC and hippocampal CA, two regions early affected by AD neuropathology, are responsible, at least partially, for associative memory deficits observed in MCI patients. PMID- 21640839 TI - Antidepressant medications reduce subcortical-cortical resting-state functional connectivity in healthy volunteers. AB - Studies have revealed abnormalities in resting-state functional connectivity in those with major depressive disorder specifically in areas such as the dorsal anterior cingulate, thalamus, amygdala, the pallidostriatum and subgenual cingulate. However, the effect of antidepressant medications on human brain function is less clear and the effect of these drugs on resting-state functional connectivity is unknown. Forty volunteers matched for age and gender with no previous psychiatric history received either citalopram (SSRI; selective serotonergic reuptake inhibitor), reboxetine (SNRI; selective noradrenergic reuptake inhibitor) or placebo for 7 days in a double-blind design. Using resting state functional magnetic resonance imaging and seed based connectivity analysis we selected the right nucleus accumbens, the right amygdala, the subgenual cingulate and the dorsal medial prefrontal cortex as seed regions. Mood and subjective experience were also measured before and after drug administration using self-report scales. Despite no differences in mood across the three groups, we found reduced connectivity between the amygdala and the ventral medial prefrontal cortex in the citalopram group and the amygdala and the orbitofrontal cortex for the reboxetine group. We also found reduced striatal-orbitofrontal cortex connectivity in the reboxetine group. These data suggest that antidepressant medications can decrease resting-state functional connectivity independent of mood change and in areas known to mediate reward and emotional processing in the brain. We conclude that hypothesis-driven seed based analysis of resting-state fMRI supports the proposition that antidepressant medications might work by normalising the elevated resting-state functional connectivity seen in depressed patients. PMID- 21640841 TI - A comparison of voxel and surface based cortical thickness estimation methods. AB - Cortical thickness estimation performed in-vivo via magnetic resonance imaging is an important technique for the diagnosis and understanding of the progression of neurodegenerative diseases. Currently, two different computational paradigms exist, with methods generally classified as either surface or voxel-based. This paper provides a much needed comparison of the surface-based method FreeSurfer and two voxel-based methods using clinical data. We test the effects of computing regional statistics using two different atlases and demonstrate that this makes a significant difference to the cortical thickness results. We assess reproducibility, and show that FreeSurfer has a regional standard deviation of thickness difference on same day scans that is significantly lower than either a Laplacian or Registration based method and discuss the trade off between reproducibility and segmentation accuracy caused by bending energy constraints. We demonstrate that voxel-based methods can detect similar patterns of group-wise differences as well as FreeSurfer in typical applications such as producing group wise maps of statistically significant thickness change, but that regional statistics can vary between methods. We use a Support Vector Machine to classify patients against controls and did not find statistically significantly different results with voxel based methods compared to FreeSurfer. Finally we assessed longitudinal performance and concluded that currently FreeSurfer provides the most plausible measure of change over time, with further work required for voxel based methods. PMID- 21640842 TI - Innovation in safety pharmacology testing. AB - This issue of the Journal of Pharmacological and Toxicological Methods (JPTM) is themed. It is the eighth in a series, arising from the Annual Safety Pharmacology Society (SPS) meeting. The SPS is now in its 10th year as an independent branch of biological sciences (distinct from pharmacology and toxicology) and is the primary forum for driving advances in safety pharmacology. The theme of the meeting and this journal issue is innovation, and the focus is non-clinical safety assessment of new chemical entity (NCEs). The content is informed by regulatory guidance documents (S7A and S7B) prior to first in human (FIH) studies. The manuscripts cover a broad spectrum of safety pharmacology topics from theory to practice, with interrogation of state-of-the-art techniques, and profiling of methods that are in development for safety assessment. Philosophical and strategic issues are addressed, with consideration of the use of novel methods for population pharmacokinetic (PK) analysis, abuse liability, electrocardiogram (ECG) analysis algorithms, in vitro cardiac slice preparations, human pluripotent stem cells, and a brief discussion regarding the assessment of changes in the QRS complex of the ECG indicative of drug-induced blockade of cardiac sodium channels. Safety pharmacology methods continue to evolve. PMID- 21640843 TI - Post-transcriptional gene regulation following exposure of osteoarthritic human articular chondrocytes to hyperosmotic conditions. AB - OBJECTIVE: Osmolarity is a major biophysical regulator of chondrocyte function. Modulation of chondrocytic marker gene expression occurs at the post transcriptional level following exposure of human articular chondrocytes (HAC) to hyperosmotic conditions. This study aims to further characterise the post transcriptional response of HAC to hyperosmolarity. METHODS: Gene expression and microRNA (miRNA) levels in freshly isolated HAC after 5h under control or hyperosmotic conditions were measured using microarrays. Regulated genes were checked for the presence of AU rich elements (AREs) in their 3' untranslated regions (3'UTR), whilst gene ontology was examined using Ingenuity Pathway Analysis (IPA). RNA decay rates of candidate ARE-containing genes were determined in HAC using actinomycin D chase experiments and the involvement of the p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinases 1 and 2 (ERK1/2) pathways were investigated using pharmacological inhibitors. RESULTS: Hyperosmolarity led to the regulation of a wide variety of genes. IPA identified enrichment of genes involved with cell stress responses, cell signalling and transforming growth factor beta (TGFbeta) signalling. Importantly, upregulated genes were over-represented with those containing AREs, and RNA decay analysis demonstrated that many of these were regulated post transcriptionally by hyperosmolarity in HAC. Analysis of miRNA levels in HAC indicated that they are only modestly regulated by hyperosmotic conditions, whilst inhibitor studies showed that p38 MAPK and ERK1/2 were able to block hyperosmotic induction of many of these genes. CONCLUSION: Through microarray and bioinformatics analysis we have identified genes which are post-transcriptionally regulated in HAC following exposure to hyperosmotic conditions. These genes have a range of functions, and their regulation involves transduction through the p38 MAPK and ERK1/2 pathways. Interestingly, our results suggest that miRNA regulation is not key to the process. Overall, this work illustrates the range of processes regulated in chondrocytes by changes in their osmotic environment, and underlines the importance of post-transcriptional mRNA regulation to chondrocyte function. PMID- 21640844 TI - Core signaling pathways of survival/death in autophagy-related cancer networks. AB - Autophagy (macroautophagy), an evolutionarily conserved lysosomal degradation process, is implicated in a wide variety of pathological processes including cancer. Autophagy plays the Janus role in regulating several survival or death signaling pathways that may decide the fate of cancer cell. Accumulating evidence has revealed the core molecular machinery of autophagy in tumor initiation and progression; however, the intricate relationships between autophagy and cancer are still in its infancy. In this review, we summarize several key survival/death pathways such as mTOR subnetwork, Beclin 1 interactome, and p53 signaling that may play the crucial roles for the regulation of the autophagy-related cancer networks. Therefore, a better understanding of the relationships between autophagy and cancer may ultimately allow cancer biologists and clinicians to harness core autophagic pathways for the discovery of potential novel drug targets. PMID- 21640846 TI - KCND3 mutations in Brugada syndrome: the plot thickens. PMID- 21640847 TI - Molecular characterization of genotype G6 human rotavirus strains detected in Italy from 1986 to 2009. AB - Group A human rotavirus (HRV) strains with a bovine-like (G6) major outer capsid protein VP7 were first detected in Palermo, Italy, in the late 1980s, and subsequently worldwide. During a 25-year rotavirus surveillance period, additional HRV G6 strains, associated with either a P[9] or P[14] VP4 genotype, have been detected sporadically, but repeatedly, in Palermo. Whether these G6 HRVs were transmitted to humans directly from an animal reservoir or could have circulated at low prevalence in susceptible individuals is uncertain. Upon sequence analyses of the VP7, VP4, VP6, NSP4 and NSP5 gene segments, all the Italian HRV strains displayed a conserved genotype constellation, G6-P[9]/[14]-I2 E2-H3. Intra-genotypic lineages and/or sub-lineages were observed among the various HRV strains, with some lineage/sublineage combinations being retained over time. Interestingly, two epidemiologically unrelated G6P[9] viruses, collected in the same rotavirus season, were found to have a clonal origin. In conclusion, our results indicate not only diverse origin of animal derived G6 HRVs in Palermo but also suggest human-to-human transmission of certain strains. PMID- 21640848 TI - Full genomic analyses of human rotavirus strains possessing the rare P[8]b VP4 subtype. AB - Rotaviruses with the P[8] VP4 genotype are a major cause of acute infantile diarrhea. The P[8] genotype is classified into two genetically distinct subtypes, P[8]a and P[8]b. Most of the P[8] strains belong to subtype P[8]a, whilst P[8]b strains are rare. To date, the whole genomes of a few P[8]a strains have been analyzed, whilst there are no reports on full genomic analysis of the P[8]b strains. To determine the genetic makeup of the rare P[8]b strains and their overall genetic relatedness to the P[8]a strains, the present study analyzed the full genomes of a human G9P[8]b strain, MMC38, and a G1P[8]b strain, MMC71, detected in Bangladesh in 2005. By nucleotide sequence identities and phylogenetic analyses, strains MMC38 and MMC71 exhibited a human rotavirus Wa like genotype constellation. Except for the VP4 gene, all the genes of strains MMC38 and MMC71 were closely related to cognate genes of the contemporary and more recent human Wa-like G1P[8]a, G9P[8]a, G11P[8]a, G11P[25], G12P[6] and G12P[8]a strains, including those from Bangladesh. Therefore, strains MMC38 and MMC71 possessed the genetically distinct P[8]b VP4 gene on a common human Wa-like genetic backbone, pointing towards their possible origin from reassortment events between common human Wa-like strains and unidentified rotavirus strains possessing the rare P[8]b-like VP4 gene. Since strains with this stable Wa-like genetic backbone can spread rapidly, and it is not certain as to whether the current rotavirus vaccines will be equally efficacious against the P[8]b strains as the P[8]a strains, proper detection of P[8]b strains and their whole genomic analyses might be of public health significance. To our knowledge, the present study is the first report on full genomic analysis of the rare P[8]b rotavirus strains. PMID- 21640849 TI - Bacillus anthracis: molecular taxonomy, population genetics, phylogeny and patho evolution. AB - Bacillus anthracis, the etiological agent of anthrax, manifests a particular bimodal lifestyle. This bacterial species alternates between short replication phases of 20-40 generations that strictly require infection of the host, normally causing death, interrupted by relatively long, mostly dormant phases as spores in the environment. Hence, the B. anthracis genome is highly homogeneous. This feature and the fact that strains from nearly all parts of the world have been analysed for canonical single nucleotide polymorphisms (canSNPs) and variable number tandem repeats (VNTRs) has allowed the development of molecular epidemiological and molecular clock models to estimate the age of major diversifications in the evolution of B. anthracis and to trace the global spread of this pathogen, which was mostly promoted by movement of domestic cattle with settlers and by international trade of contaminated animal products. From a taxonomic and phylogenetic point of view, B. anthracis is a member of the Bacillus cereus group. The differentiation of B. anthracis from B. cereus sensu stricto, solely based on chromosomal markers, is difficult. However, differences in pathogenicity clearly differentiate B. anthracis from B. cereus and are marked by the strict presence of virulence genes located on the two virulence plasmids pXO1 and pXO2, which both are required by the bacterium to cause anthrax. Conversely, anthrax-like symptoms can also be caused by organisms with chromosomal features that are more closely related to B. cereus, but which carry these virulence genes on two plasmids that largely resemble the B. anthracis virulence plasmids. PMID- 21640850 TI - Autoimmune blistering diseases of the skin. AB - Autoimmune skin diseases represent a heterogeneous group of disorders with grossly diverging clinical manifestations but partly shared underlying immunological mechanisms. They may affect the skin as an isolated organ or among systemic diseases. In addition unspecific cutaneous symptoms or drug-induced unwanted effects can be seen and have to be carefully dissected from an exacerbations of the underlying disease. Growing pathogenic knowledge has elucidated serological and clinical pictures heterogeneity and at the same time increased the therapeutic armentarium for these partly life-threatening diseases. In this review, the focus is on autoimmune bullous diseases with the skin as the major target which involve antigens of epidermis, basal membrane or dermis. Among these the pemphigoid and pemphigus group may be differentiated from dermatitis herpetiformis Duhring and epidermolysis bullosa acquisita. Interestingly, pathogenetically relevant antibody responses of IgA subtype can be found in any of the first three groups. Their clinical picture as well as therapeutic response are distinctly different from the other mainly IgG mediated subsets. Though systemic corticosteroids are still the mainstay of therapy, differential approaches using diverse adjuvant drugs are available. Immunoserological data may help characterize subsets and monitor clinical diseases as well as therapeutic response. PMID- 21640851 TI - Analysis of catalytic determinants of diaminopimelate and ornithine decarboxylases using alternate substrates. AB - Diaminopimelate decarboxylase (DAPDC) and ornithine decarboxylase (ODC) are pyridoxal 5'-phosphate dependent enzymes that are critical to microbial growth and pathogenicity. The latter is the target of drugs that cure African sleeping sickness, while the former is an attractive target for antibacterials. These two enzymes share the (beta/alpha)(8) (i.e., TIM barrel) fold with alanine racemase, another pyridoxal 5'-phosphate dependent enzyme critical to bacterial survival. The active site structural homology between DAPDC and ODC is striking even though DAPDC catalyzes the decarboxylation of a D stereocenter with inversion of configuration and ODC catalyzes the decarboxylation of an L stereocenter with retention of configuration. Here, the structural and mechanistic bases of these interesting properties are explored using reactions of alternate substrates with both enzymes. It is concluded that simple binding determinants do not control the observed stereochemical specificities for decarboxylation, and a concerted decarboxylation/proton transfer at Calpha of the D stereocenter of diaminopimelate is a possible mechanism for the observed specificity with DAPDC. PMID- 21640852 TI - Sulforaphane induces apoptosis in human hepatic cancer cells through inhibition of 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase4, mediated by hypoxia inducible factor-1-dependent pathway. AB - The anti-cancer activity of sulforaphane (SFN) has recently been investigated in several cancer cell lines, including human hepatic cancers. However, the mechanism of SFN-induced cell death in human hepatic cancer cells is still not well understood. The aim of the present work is to explore the possible mechanisms of SFN-induced apoptosis in hepatocellular carcinoma cells using proteomic analysis. A two-dimensional electrophoresis (2-DE)-based-proteomic analysis was employed for identification of possible target-related proteins of SFN-induced apoptosis. Among eleven proteins identified as regulated, we focused on the down-regulation of 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase4 (PFKFB4) protein, which has been known as a key modulator of glycolysis. We also showed that SFN down-regulated the expression of the transcriptional factor, hypoxia inducible factor-1alpha (HIF-1alpha), which strongly regulates PFKFB4 expression. In order to obtain a broad understanding of the correlation of HIF 1alpha and SFN, we observed the inhibition of the activity of mitogen-activated protein kinases, regulators of HIF-1alpha activity. Our findings suggest that SFN is a potent inducer of apoptosis in hepatocellular carcinoma cells via PFKFB4 inhibition pathways. HIF-1 pathway inhibition may be mediated by the inhibition of mitogen-activated protein kinases. PMID- 21640853 TI - Melt electrospinning of biodegradable polyurethane scaffolds. AB - Electrospinning from a melt, in contrast to from a solution, is an attractive tissue engineering scaffold manufacturing process as it allows for the formation of small diameter fibers while eliminating potentially cytotoxic solvents. Despite this, there is a dearth of literature on scaffold formation via melt electrospinning. This is likely due to the technical challenges related to the need for a well-controlled high-temperature setup and the difficulty in developing an appropriate polymer. In this paper, a biodegradable and thermally stable polyurethane (PU) is described specifically for use in melt electrospinning. Polymer formulations of aliphatic PUs based on (CH(2))(4) content diisocyanates, polycaprolactone (PCL), 1,4-butanediamine and 1,4 butanediol (BD) were evaluated for utility in the melt electrospinning process. The final polymer formulation, a catalyst-purified PU based on 1,4-butane diisocyanate, PCL and BD in a 4/1/3M ratio with a weight-average molecular weight of about 40kDa, yielded a nontoxic polymer that could be readily electrospun from the melt. Scaffolds electrospun from this polymer contained point bonds between fibers and mechanical properties analogous to many in vivo soft tissues. PMID- 21640854 TI - Three-dimensional biodegradable microscaffolding: scaffold characterization and cell population at single cell resolution. AB - Engineering artificial tissue scaffolds with a similar organization to that of the natural tissue is a key element to the successful recapitulation of function. However, three-dimensional (3-D) fabrication of tissue scaffolds containing complex microarchitectures still remains a challenge. In addition, little attention has been paid to the issue of how to incorporate cells within 3-D tissue scaffolds that contain precisely engineered architectures. Here we report a 3-D biodegradable microscaffolding (3D-BMS) technology and its process characterization as well as a microscale cellular loading technology as an efficient way to massively populate biodegradable polymers with cells at single cell resolution. In this study a particular emphasis was given to characterization of the material properties of the biodegradable polymers undergoing the 3D-BMS processes. Optimal process conditions were identified in order to avoid any unwanted change in material properties, such as crystallinity and scaffold strength, that have a direct impact on the degradation speed and physical integrity of the constructed scaffolds. For precise control of the cell distribution within the microstructured scaffolds a high precision microsieve structure was designed to localize rat hepatocytes and human articular chondrocytes in the biodegradable polymers. Cell suspensions were passed at a predetermined flow rate through biodegradable polymer layers that contained tapered microholes in a massively parallel process. This high resolution cell seeding method allows accurate manipulation of cell placement in thin layers of biodegradable polymers. PMID- 21640855 TI - The relative effects of radiation crosslinking and type of counterface on the wear resistance of ultrahigh-molecular-weight polyethylene. AB - The lifetime of total joint replacement prostheses utilizing ultrahigh-molecular weight polyethylene (UHMWPE) components has historically been determined by their wear resistance. It has been discovered that radiation crosslinking of UHMWPE can substantially increase its wear resistance. However, it is also well recognized that there is a radiation-dose-dependent decrease in several important mechanical properties of UHMWPE, such as fracture toughness and resistance to fatigue crack propagation. In this study, the effect of radiation crosslinking (followed by remelting) on the morphology, tensile properties and wear resistance of UHMWPE was investigated. Wear tests were conducted against both the commonly used cobalt chromium counterface polished to implant grade smoothness as well as a smoother ceramic (alumina) counterface. The results showed that 50kGy dose radiation crosslinking increased the wear resistance of UHMWPE against the cobalt-chromium counterface 7-fold, but the coupling of remelted, crosslinked UHMWPE against the smoother alumina counterface led to a 20-fold increase in wear resistance. This study shows that the use of an alumina counterface would circumvent the need to use a high radiation dose in crosslinking UHMWPE, associated with poor mechanical properties, without compromising wear resistance. PMID- 21640856 TI - Paediatric femoral hernia--the diagnostic challenge. AB - INTRODUCTION: Femoral herniae are uncommon in childhood and pre-operative misdiagnosis is common. This can result in increased complications or inappropriate inguinal exploration. The aim of this retrospective study was to assess a tertiary centre's experience with paediatric femoral hernia over a 12 year period. METHODS: Children who underwent femoral hernia repair at a single centre were identified from a prospectively maintained database. Casenotes were reviewed for demographic data and details of presentation, operation and recurrence. RESULTS: Sixteen children with a median age of 7 (range 3-16) years were identified. One patient developed bilateral femoral herniae. All children were referred with a groin lump but in only one instance did the referring clinician establish the diagnosis of femoral hernia. Emergency repair was required in 2 patients (12%). Eleven femoral herniae were diagnosed following clinical assessment ultrasound. The remainder were identified intra-operatively following negative inguinal exploration. Intra-operatively the femoral canal was closed with sutures (n = 16) or mesh plug (n = 1). Only one patient had a laparoscopic repair. Two other patients underwent laparoscopy to confirm bowel viability (n = 1) and for inguinal canal assessment with subsequent open femoral hernia repair (n = 1). All patients were reviewed in surgical clinic and no morbidity or hernia recurrences were reported. CONCLUSION: Femoral herniae are a diagnostic challenge and a high index of clinical suspicion is necessary. Ultrasonography or laparoscopy may be appropriate in equivocal cases. The long term results of paediatric femoral hernia surgery are excellent. PMID- 21640857 TI - Reducing the need for surgeons by reducing pollution-derived workload: is there a role for surgeons? AB - The need for additional surgical workforce personnel is likely to increase dramatically at a rate beyond our capacity to train them. As surgical training programmes cannot be rapidly expanded, this paper explores an alternative solution to the quandary, a reduction of the disease burden by a war on pollution. Highlighting the role of pollutants in increasing the surgical workload, it identifies potential roles for surgeons in the battle against pollution and draws attention to the need to research out agents which could protect humans against their carcinogenic effects. PMID- 21640858 TI - Proteomic detection of nitroproteins as potential biomarkers for cardiovascular disease. AB - Increased levels of reactive oxygen and nitrogen species are linked to many human diseases and can be formed as an indirect result of the disease process. The accumulation of specific nitroproteins which correlate with pathological processes suggests that nitration of protein tyrosine represents a dynamic and selective process, rather than a random event. Indeed, in numerous clinical disorders associated with an upregulation in oxidative stress, tyrosine nitration has been limited to certain cell types and to selective sites of injury. Additionally, proteomic studies show that only certain proteins are nitrated in selective tissue extracts. A growing list of nitrated proteins link the negative effects of protein nitration with their accumulation in a wide variety of diseases related to oxidation. Nitration of tyrosine has been demonstrated in diverse proteins such as cytochrome c, actin, histone, superoxide dismutase, alpha-synuclein, albumin, and angiotensin II. In vitro and in vivo aspects of redox-proteomics of specific nitroproteins that could be relevant to biomarker analysis and understanding of cardiovascular disease mechanism will be discussed within this review. PMID- 21640859 TI - Differences in bone metabolism between singleton pregnancy and twin pregnancy. AB - OBJECTIVE: The objective of this study was to examine the influence of twin pregnancy on calcium metabolism, including bone turnover markers and calcium regulating factors, by comparison between singleton pregnancy and twin pregnancy in women during pregnancy and puerperium in cross-sectional and longitudinal studies. METHODS: Women with singleton and twin pregnancies were recruited from the outpatient clinic of Tokushima University Hospital. In both cross-sectional and longitudinal studies, bone formation and resorption markers, mineral metabolism and calcium-regulating factors were measured at 10, 25, 30 and 36 weeks of pregnancy and at 4 days and 1 month postpartum in women with singleton and twin pregnancies. RESULTS: Urinary levels of cross-linked type I collagen N telopeptides and C-terminal telopeptides of type I collagen in women with twin pregnancy were significantly higher than those in women with singleton pregnancy and those high levels were observed earlier than those in women with singleton pregnancy. In the cross-sectional study, serum levels of bone-specific alkaline phosphatase, calcium and phosphate in women with twin pregnancy were higher and the levels of 1,25-(OH)2 vitamin D and 25-(OH) vitamin D in women with twin pregnancy were lower than those in women with singleton pregnancy. CONCLUSION: Changes in bone metabolism in women with twin pregnancy are different from those in women with singleton pregnancy. Early and large increases in bone turnover markers allow women with twin pregnancy to meet high fetal demand for calcium during pregnancy. PMID- 21640860 TI - Prefrontal cortical organization and function: implications for externalizing disorders. PMID- 21640861 TI - [31st congress of the Spanish Society of Family and Community Medicine]. PMID- 21640862 TI - Influence of root parallelism on the stability of extraction-site closures. AB - INTRODUCTION: In premolar extraction cases, root parallelism is recommended to preserve the stability of space closures. The influence of the degree of root parallelism on relapse of tooth extraction spaces has been a controversial topic in the literature. The aim of this study was to compare the angle between the long axes of the canine and the second premolar in patients with and without stability of extraction-space closures. METHODS: A sample of 56 patients, treated with 4 premolar extractions, was divided into 2 groups: group 1, consisting of 25 patients with reopening of extraction spaces; and group 2, consisting of 31 patients without reopening of extraction spaces. Panoramic radiographs of each patient were analyzed at the posttreatment and 1-year posttreatment stages. The data were statistically analyzed by using chi-square tests, t tests, analysis of variance (ANOVA), and Pearson correlation coefficients. RESULTS: The results showed that the groups did not differ regarding the angle between the canine and the second premolar, and there was no correlation between angular changes and reopening of extraction spaces, showing that dental angular changes are not determining factors for relapse, and other factors should be investigated. CONCLUSIONS: The final angle and the posttreatment changes observed in the angle between the long axes of the canine and the second premolar showed no influence on the relapse of extraction spaces. PMID- 21640863 TI - Airway volume for different dentofacial skeletal patterns. AB - INTRODUCTION: Our objective was to evaluate the nasal passage (NP) and oropharyngeal (OP) volumes of patients with different dentofacial skeletal patterns. METHODS: The study sample consisted of 140 patients (70 boys, 70 girls), divided into 3 groups as Class I (1 <= ANB <= 3), Class II (ANB > 3), and Class III (ANB < 1), and then further divided into 4 groups as SNA angle >= 80 degrees , SNA angle <80 degrees , SNB angle >=78 degrees , and SNB angle < 78 degrees to evaluate how the positional changes in the maxilla and the mandible affect the OP and NP variables. Differences between groups were determined by using the Kruskal-Wallis test. Correlations between the variables were tested with the Spearman correlation coefficient. The linear multiple regression test was applied to create a model for the airway volumes separately. RESULTS: The OP volume of the Class II subjects (n = 50) was significantly lower when compared with that of the Class I (n = 46) and Class III subjects (n = 44). The only statistically significant difference for NP volume was observed between the Class I and Class II groups. The mean OP airway volume of subjects with retruded mandibular positions was statistically significantly smaller when compared with the subjects with higher SNB angles. The area of the most constricted region at the base of the tongue (minAx) had a high potential in explaining the OP volume, whereas the NP volume models were not as successful as the OP counterpart. However, minAx was also entered into the NP volume equations as an explanatory variable. CONCLUSIONS: The OP airway volumes of Class II patients were smaller when compared with Class I and Class III patients. It was observed that mandibular position with respect to cranial base had an effect on the OP airway volume. The only significant difference for the NP volume was between the Class I and Class II groups, with a smaller volume observed for the Class II group. PMID- 21640864 TI - Evaluating craniofacial asymmetry with digital cephalometric images and cone-beam computed tomography. AB - INTRODUCTION: The aim of this study was to evaluate craniofacial asymmetry by using 2-dimensional (2D) posteroanterior cephalometric images, 3-dimensional cone beam computed tomography (CBCT), and physical measurements (gold standard). METHODS: Ten dry human skulls were assessed, and radiopaque markers were placed on 17 skeletal landmarks. Twenty linear measurements were taken on each side to compare the right and left sides and to compare these measurements with the physical measurements made with a digital caliper. To acquire the 2D posteroanterior radiographs, an Extraoral Phosphor Storage Plate (Air Techniques, Chicago, Ill) was used as the image receptor with a Eureka x-ray-Duocon Machlett unit (Machlett Laboratores, Chicago, Ill). Three-dimensional imaging data were acquired from a CB MercuRay (Hitachi Medical, Tokyo, Japan). RESULTS: On average, the right side was larger than the left for most of the 20 distances evaluated in the digital 2D and the CBCT images, and there was poor agreement between the digital 2D images and the physical measurements (kappa = 0.0609) and almost perfect agreement (kappa = 0.92) between the CBCT and physical measurements when individual measurements were considered. CONCLUSIONS: Human skulls, with no apparent asymmetry, had some differences between the right and left sides, with dominance for the right side but with no clinical significance. CBCT can better evaluate craniofacial morphology when compared with digital 2D images. PMID- 21640865 TI - Comparison between cone-beam computed tomography and intraoral digital radiography for assessment of tooth root lesions. AB - INTRODUCTION: Cone-beam computed tomography (CBCT) might be more accurate in identifying radicular surface lesions compared with digital periapical radiography. In this study, we compared these techniques in detecting simulated root resorption lesions. METHODS: A porcine mandible was used to support 10 human maxillary central incisors. CBCT and digital periapical radiographic images were generated before and after the introduction of standardized and sequentially larger root defects on either the mesial or the lingual root surfaces. The images were randomly labeled and evaluated by 3 examiners. Each image was classified according to defect size (0, none; 1, mild; 2, moderate; 3, severe). RESULTS: Interrater reliability was acceptable (0.856 <= P <= 0.981). The location of the root defect (mesial vs lingual) had no significant effect on the evaluation of defect size. Both periapical radiographs and CBCT were slightly better at detecting lingual defects than mesial defects (75% vs 65% and 65% vs 60%, respectively), but these effects were not statistically significant (P = 0.49 and P = 0.74, respectively). The mean percentages of correctly diagnosed defect sizes were 65% for CBCT and 75% for periapical radiographs. Examiners using CBCT images tended to overestimate defect sizes (kappa = 0.481) and correctly categorized teeth with no, mild-moderate, and severe defects 80%, 45%, and 90% of the time, respectively. Examiners using periapical radiographs tended to underestimate defect sizes (kappa = 0.636) and categorized teeth with no, mild-moderate, and severe defects 100%, 50%, and 100% of the time, respectively. CONCLUSIONS: There was no difference in accuracy of identifying defects between periapical radiographs and CBCT images. PMID- 21640866 TI - In-vitro study of the effect of casein phosphopeptide amorphous calcium fluoride phosphate on iatrogenic damage to enamel during orthodontic adhesive removal. AB - INTRODUCTION: White-spot lesions (WSL) might be susceptible to mechanical damage during orthodontic bracket and adhesive removal. The aims of this in-vitro study were to investigate enamel loss on bracket and adhesive removal when the brackets were surrounded by WSL and to determine the effect of remineralizing these lesions with a 1% (w/v) casein phosphopeptide amorphous calcium fluoride phosphate (CPP-ACFP) solution before bracket and adhesive removal. METHODS: Precoated metal mandibular incisor brackets were centrally bonded onto polished third molars and WSL produced by exposure to a demineralization buffer for 4, 12, and 30 days (n = 20 per group). Half of the demineralized window was covered with acid-resistant nail varnish, and the specimens were then subjected to remineralization with 1% CPP-ACFP. Brackets and residual adhesive were removed, and enamel damage was assessed by digital photography, profilometry, and scanning electron microscopy. Lesion depth, mineral loss, and remineralization were measured by transverse microradiography. RESULTS: WSL enamel around the bracket was more susceptible to iatrogenic damage at adhesive removal compared with sound enamel. Remineralization of lesions with 1% CPP-ACFP before adhesive removal significantly (P <0.002) reduced the area and depth of damage. CONCLUSIONS: Remineralizing WSLs with CPP-ACFP before adhesive removal reduced iatrogenic enamel damage. PMID- 21640867 TI - Osteocalcin and N-telopeptides of type I collagen marker levels in gingival crevicular fluid during different stages of orthodontic tooth movement. AB - INTRODUCTION: The aim of the study was to quantitatively and qualitatively analyze changes in the gingival crevicular fluid (GCF) during orthodontic movement by assessing the levels of cross-linked N-telopeptides of type I collagen marker (NTx) and osteocalcin (OC) activity. METHODS: Twenty patients requiring all first premolars to be extracted were selected and treated with conventional straight-wire mechanotherapy. The canines were retracted with closed coil springs. The maxillary canine on one side acted as the experimental site, and the contralateral canine was the control. GCF was collected from around the canines before retraction, and 1 hour, 1 day, 7 days, 14 days, and 21 days after retraction. GCF NTx and OC levels were estimated and compared with the control side. RESULTS: The results showed statistically significant changes in NTx and OC levels on days 7, 14, and 21 when we compared the experimental and control sides. The peak in all activity of the variables occurred on day 14 after retraction. CONCLUSIONS: The study showed that NTx and OC levels can be successfully estimated in the GCF, and its increased levels might indicate the active tooth movement phase in orthodontic therapy. PMID- 21640868 TI - Create realistic objectives. PMID- 21640869 TI - The apple of Sir Isaac Newton. PMID- 21640871 TI - Dental development in children with hypodontia. PMID- 21640872 TI - Maxillary position. PMID- 21640875 TI - Inappropriate liaisons. PMID- 21640877 TI - Are orthodontic distalizers reinforced with the temporary skeletal anchorage devices effective? AB - INTRODUCTION: Our objective was to perform a systematic review of studies pertaining to the distalization of teeth with appliances reinforced with temporary skeletal anchorage devices. METHODS: PubMed, Embase, Cochrane Central Register of Controlled Trials, Web of Knowledge, Ovid, and Scopus were searched until the second week of August 2010 to identify all articles reporting on the use of orthodontic implants or miniplates in distalization of teeth. The quality of the relevant studies was ranked on an 11-point scale, from low to high quality. RESULTS: Twelve relevant articles were identified. The distal movement of the maxillary molars was from 3.3 to 6.4 mm; the concomitant molar distal tipping was from 0.80 degrees to 12.20 degrees . The maxillary incisors remained stable during molar distalization. The assessment of study quality showed that 8 studies were of low and 4 of medium quality. CONCLUSIONS: Molar distalizers reinforced with the temporary skeletal anchorage devices seem to effectively move molars distally without unwanted mesial incisor tipping. Because of the lack of high-quality studies, however, the findings of this study should be interpreted with caution. PMID- 21640878 TI - Ion release from orthodontic brackets in 3 mouthwashes: an in-vitro study. AB - INTRODUCTION: Stainless steel orthodontic brackets can release metal ions into the saliva. Fluoridated mouthwashes are often recommended to orthodontic patients to reduce the risk of white-spot lesions around their brackets. However, little information is available regarding the effect of different mouthwashes in ion release of orthodontic brackets. The purpose of this study was to measure the amount of metal ion release from orthodontic brackets when kept in different mouthwashes. METHODS: One hundred sixty stainless steel brackets (0.022-in, 3M Unitek, Monrovia, Calif) were divided randomly into 4 equal groups and immersed in Oral B (Procter & Gamble, Weybridge, United Kingdom), chlorhexidine (Shahdaru Labratories, Tehran, Iran), and Persica (Poursina Pharmaceutical Laboratories, Tehran, Iran) mouthwashes and distilled deionized water and incubated at 37 degrees C for 45 days. Nickel, chromium, iron, copper, and manganese released from the orthodontic brackets were measured with an inductively coupled plasma spectrometer. For statistical analysis, 1-way analysis of variance (ANOVA) and the Duncan multiple-range tests were used. RESULTS: The results showed that ion release in deionized water was significantly (P <0.05) higher than in the 3 mouthwashes. Higher ion release was found with chlorhexidine compared with the other 2 mouthwashes. There was no significant difference (P >0.05) in nickel, chromium, iron, and copper ion release in the Oral B and Persica mouthwashes. The level of manganese release was significantly different in all 4 groups. CONCLUSIONS: If ion release is a concern, Oral B and Persica mouthwashes might be better options than chlorhexidine for orthodontic patients with stainless steel brackets. PMID- 21640879 TI - Longitudinal changes in gingival crevicular fluid after placement of fixed orthodontic appliances. AB - INTRODUCTION: Bacterial plaque is an etiologic factor in the development of gingival inflammation and periodontitis. The presence of orthodontic bands and brackets influences plaque growth and maturation. The purposes of this research were to monitor microbiologic and periodontal changes after placement of orthodontic attachments over a 1-year period and to link these changes to alterations in cytokine concentrations in the gingival crevicular fluid (GCF). METHODS: This longitudinal split-mouth trial included 24 patients. Supragingival and subgingival plaque composition, probing depth, bleeding on probing, and GCF flow and composition were assessed at baseline (Tb) and after 1 year (T52). A statistical comparison was made over time and between the banded and bonded sites. Prognostic factors for the clinical reaction at T52 in the GCF at Tb were determined. RESULTS: Between Tb and T52, the pathogenicity of the plaque and all periodontal parameters increased significantly, but intersite differences were not seen, except for bleeding on probing. The cytokine concentrations in the GCF did not differ significantly between the sites or between Tb and T52. The interleukin-6 concentration in the GCF at Tb was a significant predictive value for the GCF flow at T52 (P <0.05). The same relationship was found between the interleukin-8 concentration at Tb and the increase in probing depth at T52 (P <0.05). CONCLUSIONS: Interleukin-6 and interleukin-8 concentrations before orthodontic treatment were shown to be significant predictive factors for some potential inflammatory parameters during treatment. PMID- 21640880 TI - How does tooth eruption relate to vertical mandibular growth displacement? AB - INTRODUCTION: Our objectives were to investigate the eruptive patterns of the mandibular teeth and assess their associations with mandibular growth displacements. METHODS: Cephalograms for a mixed-longitudinal sample of 124 French-Canadian girls were evaluated between 10 and 15 years of age. Vertical mandibular displacement and mandibular eruption were evaluated by using cranial and mandibular superimpositions, respectively. Multilevel modeling procedures were used to estimate each subject's growth change over time. Stepwise multiple regressions were used to determine the amount and relative magnitudes of variations in mandibular eruption explained by mandibular growth displacement, controlling for vertical maxillary tooth movements. RESULTS: Cubic polynomial models explained between 91% and 98% of the variations in eruption and vertical growth displacement. All curves showed acceleration of eruption until approximately 12 years of age, after which eruption decelerated. The eruption of the mandibular teeth demonstrated greater relative variability than did vertical mandibular growth displacements. Independent of the overall movements of the maxillary molars, inferior mandibular growth displacement explained approximately 54% of the variation in mandibular molar eruption between 10.5 and 14.5 years of age. CONCLUSIONS: Inferior mandibular growth displacement and dental eruption followed similar patterns of change during adolescence. Based on their associations and the differences in variability identified, mandibular eruption appears to compensate for or adapt to growth displacements. PMID- 21640881 TI - Histologic evaluation of root-surface healing after root contact or approximation during placement of mini-implants. AB - INTRODUCTION: Placement of mini-implants carries with it the risk of iatrogenic damage to the adjacent root surfaces. The aims of this study were to assess the type of trauma incurred on tooth roots after contact or approximation with mini implants during placement, and to observe and analyze the healing responses via histologic analysis. METHODS: Four male minipigs were used as experimental subjects. Twenty mini-implants (1.6 * 8 mm) were implanted in each minipig into the buccal sides of all 4 quadrants between the roots of teeth so that contact or approximation between the mini-implant and root surface occurred, with the aid of dental fluoroscopy. All mini-implants on the left side of the mouth were left in situ, and all mini-implants on the right side were removed immediately after placement. The minipigs were killed at 4-week intervals up to week 16, and histologic sections were made. RESULTS: When mini-implants were left in situ, the root surface was mostly resorbed away from the mini-implant thread. Partial repair started at 8 weeks. When the mini-implant thread was left touching the root, there was no normal healing response. If the mini-implant was placed less than 1 mm from the periodontal ligament, resorption was evident on the root surface. Abnormal healing responses were seen when the pulp tissue was ruptured, mostly through osteodentin formation. In all instances after mini-implant removal immediately after placement, varying degrees of cementum repair was observed. CONCLUSIONS: Immediate removal of the mini-implant leads to cementum repair, whereas leaving the mini-implant in place will cause either a delay in repair or no repair. Placing mini-implants less than 1 mm from the root surface causes root surface resorption. PMID- 21640882 TI - Clinical evaluation of fiber-reinforced-plastic bonded orthodontic retainers. AB - INTRODUCTION: This clinical study was designed to evaluate the efficacy of innovative fiber-reinforced-thermoplastic (FRP) bonded orthodontic retainers. METHODS: Anterior lingual retainers were formed by using a 2-step process from preimpregnated unidirectional long glass fibers (volume fraction, 0.25) in a thermoplastic resin matrix of either poly(ethylene terephthalate glycol) or polycarbonate. Seventy-six canine-to-canine retainers were placed in 56 patients by using the acid-etch technique over a 34-month period. They were evaluated for clinical acceptability to function as a retainer, structural integrity of the FRP, and integrity of the bonding. Variables examined included material composition, design factors, and mechanism of failure of the retainers. The results were analyzed by using the univariate Kaplan-Meier survival method and the multivariate Cox regression model. RESULTS: The overall median survival time was 7.6 months, with 33% surviving after 12 months. The retainer with the longest service was in use for over 24 months. The variables with the most significant effects and improved survival rates were FRP formulation (polycarbonate), retainer thickness (1.02 mm), and number of teeth overlapped (all 6 teeth). Failure was primarily due to bond failure at the enamel-adhesive or adhesive-FRP interface. CONCLUSIONS: With improved survival times, FRP retainers could be a viable alternative to metal retainers. PMID- 21640883 TI - Stability and relapse of maxillary anterior crowding treatment in class I and class II Division 1 malocclusions. AB - INTRODUCTION: The maxillary anterior teeth are the most important to facial esthetics because they are the first to show on a smile. Therefore, stability of the maxillary anterior teeth alignment is an important issue. The objective of this study was to compare the stability of maxillary anterior tooth alignment in Class I and Class II Division 1 malocclusions. METHODS: The sample comprised dental casts of 70 patients with Class I and Class II Division 1 malocclusions and a minimum of 3 mm of maxillary anterior crowding measured by an irregularity index. The patients were treated with extractions and evaluated at pretreatment and posttreatment and at least 5 years after treatment. The sample was divided into 3 groups: group 1, Class I malocclusion treated with 4 first premolar extractions comprising 30 subjects, with an initial age of 13.16 years and 8.59 mm of initial maxillary irregularity; group 2, Class II malocclusion treated with 4 first premolar extractions comprising 20 subjects, with an initial age of 12.95 years and 11.10 mm of maxillary irregularity; and group 3, Class II malocclusion treated with 2 first maxillary premolar extractions comprising 20 subjects, with an initial age of 13.09 years and 9.68 mm of maxillary irregularity. RESULTS: The decrease in the maxillary irregularity index was significantly greater in group 2 than in group 1 during treatment. The stability of maxillary anterior alignment was 88.12% over the long term; 77% of the linear displacement of the anatomic contact points tended to return to their original positions. CONCLUSIONS: Stability of maxillary anterior alignment between the 3 groups was similar. The stability of maxillary anterior alignment was high over the long term, but a high percentage of teeth tended to return to their original positions. PMID- 21640884 TI - Influence of different modeling strategies for the periodontal ligament on finite element simulation results. AB - INTRODUCTION: The finite element method is a promising tool to investigate the material properties and the structural response of the periodontal ligament (PDL). To obtain realistic and reproducible results during finite element simulations of the PDL, suitable bio-fidelic finite element meshes of the geometry are essential. METHODS: In this study, 4 independent coworkers generated altogether 17 volume meshes (3-dimensional) based on the same high-resolution computed-tomography image data set of a tooth obtained in vivo to compare the influence of the different model generation techniques on the predicted response to loading for low orthodontic forces. RESULTS: It was shown that the thickness of the PDL has a significant effect on initial tooth mobility but only a remarkably moderate effect on the observed stress distribution in the PDL. Both the tooth and the bone can be considered effectively rigid when exploring the response of the PDL under low loads. The effect of geometric nonlinearities could be neglected for the applied force system. CONCLUSIONS: Most importantly, this study highlights the sensitivity of the finite element simulation results for accurate geometric reconstruction of the PDL. PMID- 21640885 TI - Comparison of oral impacts experienced by patients treated with labial or customized lingual fixed orthodontic appliances. AB - INTRODUCTION: Our objective was to compare the oral impacts experienced by patients treated with labial or customized lingual fixed orthodontic appliances. METHODS: This was an age- and sex-matched prospective longitudinal study of 60 adult patients treated with either labial or customized lingual fixed orthodontic appliances over a 3-month period. Ratings of oral impacts experienced and satisfaction were made on visual analog scales at 3 time points after appliance fixation. Variations in oral impacts and satisfaction over the trajectory of treatment were assessed. Area-under-the-curve analyses were conducted to assess variations in oral impacts and satisfaction between the groups. RESULTS: All patients experienced oral impact disturbances, although these disturbances decreased over time (P < 0.001). Patients treated with customized lingual appliances reported more oral discomfort (P < 0.001), dietary changes (P < 0.001), swallowing difficulty (P < 0.001), speech disturbances (P < 0.001), and social problems (P < 0.001) than did those in the other group. There was no significant difference between the groups regarding ratings of oral self-care, mastication, and satisfaction level of treatment (P > 0.05). CONCLUSIONS: The findings indicate that oral impacts are commonly experienced during both labial and customized lingual fixed orthodontic therapies. However, the oral impacts decreased over the observational period. Patients treated with customized lingual appliances experienced more oral impacts. Both groups had similar levels of treatment satisfaction. PMID- 21640886 TI - Development of a measure for orthodontists to evaluate patient compliance. AB - INTRODUCTION: Interruption of treatment and poor compliance are problems in orthodontics, especially when the patient does not pay for treatment. The aim of this study was to develop a measure for orthodontists, regardless of type of practice, to assess their opinions about patient compliance. METHODS: A questionnaire, based on an orthodontic patient cooperation scale, was modified in 2 pilot phases. The piloted version was tested among 249 respondents. A principal component analysis was performed that included factors with an eigenvalue greater than 1. Reliability was assessed by means of internal consistency with Cronbach's alphas and by test-retest (n = 40) measures, using an intraclass correlation coefficient. To assess construct validity, the responses of private and public practitioners were analyzed with chi-square and t tests. RESULTS: The response rate was 77%. The final questionnaire showed good reliability: Cronbach's alpha coefficient was 0.878, and the intraclass correlation coefficient after the test retest was 0.868. The participation rate was good, there were few missing values, and the study was multifaceted; thus the questionnaire also showed good validity for face, content, and construct. The differences between private and public practitioners were statistically significant. Five factors describing the different aspects of compliance were shown. CONCLUSIONS: Observed differences between public and private orthodontists and dentists performing orthodontics indicated clearly the need to develop a specific measure for countries with a publicly funded system of oral health care. The measure showed good reliability and validity for face, content, and construct among Finnish orthodontists and dentists performing orthodontics. The predictive validity of the measure to assess actual patient compliance remains to be tested. PMID- 21640887 TI - Vertical craniofacial growth changes in French-Canadians between 10 and 15 years of age. AB - INTRODUCTION: Because of limited available reference data, this study described the vertical growth changes that occur in untreated adolescents 10 to 15 years of age and evaluated the validity of measurements commonly used to classify patients' vertical growth tendencies. METHODS: The sample consisted of 228 subjects (119 boys, 109 girls) between 10 and 15 years of age with normal occlusions or malocclusions who had lateral cephalograms (n = 1303) taken annually. Based on 6 landmarks, 3 angles (PPA, MPA, PP/MPA) and 2 proportions (PFH:AFH and UFH:LFH) were calculated. To reduce errors, each subject's growth curve was estimated by using multilevel modeling procedures, and the estimated values were analyzed. RESULTS: Growth changes between 10 and 15 years for each of the 5 measurements followed relatively simple (linear or quadratic) polynomial models. On average, PPA and PFH:AFH increased, and MPA and PP/MPA decreased. The UFH:LFH ratio increased during the first few years and then decreased. MPA, PP/MPA, and PFH:AFH showed moderately high intercorrelations; PPA displayed moderate to moderately low correlations with UFH:LFH; UFH:LFH showed a moderate correlation with PP/MPA. Approximately 75% to 86% of the subjects classified as hyperdivergent or hypodivergent at 10 years maintained their classification. Subjects classified as hyperdivergent at 15 years of age showed significantly greater growth changes than did those classified within normal limits, who, in turn, showed greater changes than did the hypodivergent subjects. CONCLUSIONS: Measurements typically used to classify vertical growth tendencies changed significantly during adolescence, with boys generally showing greater changes than girls. Although MPA, PFH:AFH, and PP/MPA measured the same phenotypic attribute, PPA and UFH:LFH were relatively independent of the other 3 measurements. Most subjects maintained their vertical facial types, but some worsened, and others improved. PMID- 21640888 TI - Esthetic preferences for the shape of anterior teeth in a posed smile. AB - INTRODUCTION: Although various aspects of smile esthetics have been considered in many studies, few of them have compared laypeople's preferences for an esthetic smile with regard to the shape of the anterior teeth. The aim of this study was to evaluate the effects of the shapes of the maxillary anterior teeth in men and women on the perception of an esthetic smile by different age and sex groups of laypeople. METHODS: Two pamphlets, each consisting of 9 smile images that differed only in the shapes of the canines and incisors, were shown to 4 groups of judges of different ages and sexes. They were asked to rank feminine and masculine smile images according to their preferences using a 10-cell linear horizontal visual analog scale. RESULTS: Neither in the men's nor in the women's pamphlets were there significant differences between the scores of the 4 groups of judges to the various tooth forms. In both pamphlets, regardless of canine shape, all groups of judges gave significantly different scores to square, square round, and round incisors so that the median of scores for square incisors was significantly lower than those of the 2 other incisor forms. CONCLUSIONS: Sex and age of the laypersons did not affect their esthetic perception of the shapes of the maxillary anterior teeth. Incisor shape was the key determinant of their esthetic preferences; round incisors were the most esthetic. It is recommended to improve smile esthetics by mildly rounding the mesial and distal corners of square incisors. PMID- 21640889 TI - Short-term and long-term stability of surgically assisted rapid palatal expansion revisited. AB - INTRODUCTION: The purpose of this article is to present further longitudinal data for short-term and long-term stability, following up our previous article in the surgery literature with a larger sample and 2 years of stability data. METHODS: Data from 38 patients enrolled in this prospective study were collected before treatment, at maximum expansion, at removal of the expander 6 months later, before any second surgical phase, at the end of orthodontic treatment, and at the 2-year follow-up, by using posteroanterior cephalograms and dental casts. RESULTS: With surgically assisted rapid palatal expansion (SARPE), the mean maximum expansion at the first molar was 7.60 +/- 1.57 mm, and the mean relapse was 1.83 +/- 1.83 mm (24%). Modest relapse after completion of treatment was not statistically significant for all teeth except for the maxillary first molar (0.99 +/- 1.1 mm). A significant relationship (P < 0.0001) was observed between the amount of relapse after SARPE and the posttreatment observation. At maximum, a skeletal expansion of 3.58 +/- 1.63 mm was obtained, and this was stable. CONCLUSIONS: Skeletal changes with SARPE were modest but stable. Relapse in dental expansion was almost totally attributed to lingual movement of the posterior teeth; 64% of the patients had more than 2 mm of dental changes. Phase 2 surgery did not affect dental relapse. PMID- 21640890 TI - Total distalization of the maxillary arch in a patient with skeletal Class II malocclusion. AB - In nongrowing patients with skeletal Class II malocclusion, premolar extraction or maxillary molar distalization can be used as camouflage treatment. Orthodontic miniscrew implants are widely used for this purpose because they do not produce undesirable reciprocal effects and do not depend on the patient's cooperation. This article reports on maxillary molar distalization by using miniscrew implants to correct a Class II problem. The main considerations of molar distalization treatment with miniscrew implants are discussed. PMID- 21640891 TI - Transposition of a maxillary canine and a lateral incisor and use of cone-beam computed tomography for treatment planning. AB - This report describes the orthodontic treatment of a 12-year-old girl with transposition of the maxillary left canine and the lateral incisor. Cone-beam computed tomography was used during treatment planning. The transposed tooth positions were corrected with an unconventional orthodontic approach. Treatment alternatives and their clinical concerns are presented. PMID- 21640892 TI - Maxillary osteosarcoma in a young patient undergoing postorthodontic treatment follow-up: the importance of ongoing oral examinations. AB - Osteosarcoma is a common primary malignant tumor in long bones; it generally occurs in young adults. It is considered infrequent in the head and neck regions, where it is usually associated with poor outcomes and rates of survival. From a histopathologic point of view, osteosarcomas are commonly classified as osteoblastic, chondroblastic, or fibroblastic, although several unusual microscopic subtypes have also been reported. The purpose of this article was to present a case report of a maxillary chondroblastic osteosarcoma involving the maxillary sinus and the maxilla of a young woman who was diagnosed during early postorthodontic treatment follow-up. Treatment, prosthetic rehabilitation, and follow-up details are provided. Most importantly, this patient shows the importance of complete and systematic oral examinations during any routine dental treatment. PMID- 21640893 TI - Replacing a failed mini-implant with a miniplate to prevent interruption during orthodontic treatment. AB - INTRODUCTION: When mini-implants fail during orthodontic treatment, there is a need to have a backup plan to either replace the failed implant in the adjacent interradicular area or wait for the bone to heal before replacing the mini implant. We propose a novel way to overcome this problem by replacement with a miniplate so as not to interrupt treatment or prolong treatment time. METHODS: The indications, advantages, efficacy, and procedures for switching from a mini implant to a miniplate are discussed. Two patients who required replacement of failed mini-implants are presented. In the first patient, because of the proximity of the buccal vestibule to the mini-implant, it was decided to replace the failed mini-implant by an I-shaped C-tube miniplate. In the second patient, radiolucencies were found around the failed mini-implants, making the adjacent alveolar bone unavailable for immediate placement of another mini-implant. In addition, the maxillary sinus pneumatization was expanded deeply into the interradicular spaces; this further mandated an alternative placement site. One failed mini-implant was examined under a scanning electron microscope for bone attachment. RESULTS: Treatment was completed in both patients after replacement with miniplates without interrupting the treatment mechanics or prolonging the treatments. Examination under the scanning electron microscope showed partial bone growth into the coating pores and titanium substrate interface even after thorough cleaning and sterilization. CONCLUSIONS: Replacement with a miniplate is a viable solution for failed mini-implants during orthodontic treatment. The results from microscopic evaluation of the failed mini-implant suggest that stringent guidelines are needed for recycling used mini-implants. PMID- 21640894 TI - Litigation and legislation. Limits to the standard of care. PMID- 21640895 TI - KASH and SUN proteins. PMID- 21640896 TI - Microsaccades. PMID- 21640897 TI - Plants. PMID- 21640898 TI - Root development: cytokinin transport matters, too! AB - Unlike the plant hormone auxin, the mechanism and function of cytokinin transport is poorly characterised. Two new studies now demonstrate that cytokinins transported from shoot to roots via the phloem are critical for creating mutually exclusive auxin and cytokinin signalling domains that control root vascular patterning. PMID- 21640899 TI - Neuroanatomy: uninhibited connectivity in neocortex? AB - The mouse neocortex is now the focus of research using twenty-first century techniques of circuit analyses, which are revealing different wiring strategies for excitatory and inhibitory connections and providing important insights into the possible computations of cortical circuits. PMID- 21640900 TI - Force generation: ATP-powered proteasomes pull the rope. AB - Recently, single-molecule force spectroscopy techniques have provided unprecedented opportunities to apply and to quantify forces that guide protein (un-)folding. A new study provides fascinating insights into the sophisticated mechanism by which an ATP-fueled proteolytic machine generates mechanical forces to unfold and translocate multidomain substrates. PMID- 21640901 TI - Epithelial organization: new perspective on alpha-catenin from an ancient source. AB - Cadherins and catenins evidently partnered at the dawn of the animal kingdom to enable the first polarized epithelium, and perhaps animal evolution itself. New evidence from a primitive slime mold, however, suggests that alpha- and beta catenins may have engaged this function independently, long before cadherins arrived on the scene. PMID- 21640902 TI - Comparative cognition: comparing human and monkey memory. AB - Humans store a limited number of items in short-term working memory to perform subsequent operations. A newly described assessment of memory in rhesus monkeys suggests qualitative similarities and quantitative dissimilarities to humans. PMID- 21640903 TI - Primary cilia: how to keep the riff-raff in the plasma membrane. AB - A recent report suggests that plasma membrane proteins are excluded from primary cilia via anchoring to the cortical actin cytoskeleton. These findings challenge the existence of a diffusion barrier at the base of the cilium. PMID- 21640904 TI - Cognitive neuroscience: scene layout from vision and touch. AB - Parahippocampal and retrosplenial cortices respond strongly to visual scenes. A new study shows that these regions also activate when scenes are perceived haptically - even in the blind. PMID- 21640905 TI - Olfactory neuroscience: beyond the bulb. AB - High-resolution tracing of projections from the olfactory bulb to its cortical targets revealed coarse topography and stereotopy in some areas but highly distributed, combinatorial connectivity in others. These results provide a basis for understanding innate and associative olfactory processing and perception. PMID- 21640906 TI - KNL1/Spc105 recruits PP1 to silence the spindle assembly checkpoint. AB - The spindle assembly checkpoint (SAC) delays anaphase onset until kinetochores accomplish bioriented microtubule attachments [1]. Although several centromeric and kinetochore kinases, including Aurora B, regulate kinetochore-microtubule attachment and/or SAC activation [2-4], the molecular mechanism that translates bioriented attachment into SAC silencing remains unclear [5]. Employing a method to rapidly induce exact gene replacement in budding yeast [6], we show here that the binding of protein phosphatase 1 (PP1/Glc7) to the evolutionarily conserved RVSF motif of the kinetochore protein Spc105 (KNL1/Blinkin/CASC5) is essential for viability by silencing the SAC, while it plays an auxiliary nonessential role for physical chromosome segregation. Although Aurora B may inhibit this binding, persistent PP1-Spc105 interaction does not affect chromosome segregation and is insufficient to silence the SAC in the absence of microtubules, indicating that dynamic regulation of this interaction is dispensable. However, the amount of PP1 targeted to kinetochores must be finely tuned, because recruitment of either no or one extra copy of PP1 to Spc105 is detrimental, illustrating the vital impact of targeting an exiguous fraction of PP1 to the kinetochore. We propose that the PP1-Spc105 interaction enables local regulation of dynamic phosphorylation and dephosphorylation at the kinetochore to couple microtubule attachment and SAC silencing. PMID- 21640907 TI - Signaling pathways in cranial chondrosarcoma: potential molecular targets for directed chemotherapy. AB - Cranial chondrosarcoma is an indolent tumor of the skull base, which is difficult to treat due to its inaccessible location. The current mainstay of treatment is surgical resection followed by adjuvant radiation therapy. To date, chemotherapy has been largely ineffective for chondrosarcoma due to a lack of targeted therapies. This review highlights numerous active signaling pathways that have been described in human chondrosarcoma. A limited number of functional experiments suggest that integrin activation at the cell surface results in upregulation of matrix metalloproteinases and extracellular matrix degradation, leading to increased tumor cell migration. This pathway appears to be dependent on phosphoinositide-3 kinase and MEK-extracellular signal-regulated kinase (ERK) signaling. Additionally, chondrosarcoma cell proliferation and degradation is dependent on peroxisome proliferator-activated receptor-gamma (PPAR-gamma) activity, with a loss of PPAR-gamma expression and associated apoptosis in high grade tumors. The data suggest that targeting these pathways may improve control of cranial chondrosarcoma and decrease the need for hazardous recurrent operations. PMID- 21640908 TI - The tumor biology and molecular characteristics of medulloblastoma identifying prognostic factors associated with survival outcomes and prognosis. AB - Medulloblastomas (MB) are highly aggressive primitive neuroectodermal tumors (PNET) usually located in the posterior fossa. Current treatment for MBs, which includes a combination of surgery, chemotherapy and radiation, remain challenging especially in younger patients. However, advances in the understanding of regulatory pathways in cerebellar development have elucidated possible areas of dysfunction involved in tumorigenesis. Multiple studies have demonstrated the importance of the sonic hedgehog, Wnt, and Notch pathways in MB pathogenesis at the molecular level. While staging and prognosis are often based on the Chang classification system, future algorithms will involve identifying molecular markers in order to allow for more specific risk stratifications of various MB subtypes and provide improved correlation with staging and prognosis. Future development of novel therapies that target the heterogeneity of MB and are tailored to the tumor's unique molecular profile may yield improved outcomes for these patients. PMID- 21640910 TI - Surgical oncology in the community cancer center. Preface. PMID- 21640911 TI - Building a community-based cancer center program. AB - Developing a successful cancer center within the community is achievable. This article provides an understanding of the standards and guidelines of the Commission on Cancer (CoC), the different community cancer center program categories, and the accreditation process. The pivotal roles of institutional support and physician leadership in the development of a successful cancer center have been elucidated. PMID- 21640912 TI - Breast conservation therapy versus mastectomy in the community-based setting: can this rate be used as a benchmark for cancer care? AB - Despite strong evidence supporting the use of breast conservation therapy (BCT) in the treatment of breast carcinoma, the actual rates of use remain low. This article is a retrospective review of a sample of patients from the cancer registry of the Carolinas Medical Center (CMC), comparing breast conservation and mastectomy rates during an 11-year period. BCT rates have increased in CMC during this time frame and have reached national levels. Further research is needed to determine whether BCT rates can be used as a benchmark for the care of patients with cancer. PMID- 21640913 TI - American College of Surgeons Oncology Group and the community surgeon. AB - The American College of Surgeons Oncology Group (ACOSOG) conducts cancer trials that are relevant to surgeons who treat patients with breast, thoracic, and gastrointestinal cancers. ACOSOG is funded by the National Cancer Institute and is charged with conducting prospective clinical trials that address important questions in academic and community practice settings. Examples include role of axillary dissection for microscopic nodal disease, neoadjuvant therapy for organ conserving surgery, laparoscopic rectal cancer resection, mediastinal nodal staging, and sublobar resection for early-stage non-small cell lung cancer. Such trials are relevant to most practicing surgeons. PMID- 21640914 TI - A community hospital clinical trials program: infrastructure for growth. AB - This article discusses the growth and infrastructure of a clinical trials program in a large community hospital. An association between increased patient accrual and establishment of site-specific tumor boards and patient care navigators is also discussed. Strong hospital administrative support and clinical research staff are crucial for success. PMID- 21640915 TI - Hepatic tumor ablation: application in a community hospital setting. AB - Primary liver tumors are a common clinical problem in the United States and worldwide. Resection has historically been used to treat liver lesions. Commonly used liver-directed therapies include transarterial chemoembolization, selective internal radiation therapy, and ablative therapy. Only ablative therapy can cause direct destruction of the targeted tissue. The commercially available modalities in the United States are all based on thermoablative technology. This article examines the various ablative technologies and their application, as well as how these procedures can be performed safely and with optimal outcomes, in a community cancer center. PMID- 21640916 TI - Molecular analysis of breast sentinel lymph nodes. AB - Lymphatic mapping and sentinel lymph node (SLN) biopsy have become the standard of care for staging the axilla in patients with invasive breast cancer. Current histologic methods for SLN evaluation have limitations, including subjectivity, limited sensitivity, and lack of standardization. The discovery of molecular markers to detect metastases has been reported over the last 2 decades. The authors review the historical development of these markers and the clinical use of one of the molecular platforms in 478 patients at their institution. Controversies and future directions are discussed. PMID- 21640917 TI - Pancreatic resection in a large tertiary care community-based hospital: building a successful pancreatic surgery program. AB - Pancreatic resection can be performed safely in the community-based hospital setting only when appropriate systems are in place for patient selection and preoperative, operative, and postoperative care. Pancreatic surgery cannot be performed optimally without considerable investment in, and coordination of, multiple departments. Delivery of high-quality pancreatic cancer care demands a rigorous assessment of the hospital structure and the processes through which this care is delivered; however, when a hospital makes the considerable effort to establish the necessary systems required for delivery of quality pancreatic cancer care, the community and hospital will benefit substantially. PMID- 21640918 TI - Maximizing rectal cancer results: TEM and TATA techniques to expand sphincter preservation. AB - Rectal cancer management benefits from a multidisciplinary approach involving medical and radiation oncology as well as surgery. Presented are the current dominant issues in rectal cancer management with an emphasis on our treatment algorithm at the Lankenau Medical Center. By basing surgical decisions on the downstaged rectal cancer we explore how sphincter preservation can be extended even for cancers of the distal 3 cm of the rectum. TATA and TEM techniques can be used to effectively treat cancer from an oncologic standpoint while maintaining a high quality of life through sphincter preservation and avoidance of a permanent colostomy. We review the results of our efforts, including the use of advanced laparoscopy in the surgical management of low rectal cancers. PMID- 21640919 TI - Minimally invasive esophagectomy in the community hospital setting. AB - We report our initial experience with minimally-invasive esophagectomy in 32 patients at Carolinas Medical Center, a community academic medical center. Indications for surgery were adenocarcinoma in 27, squamous cell carcinoma in 3, and benign stricture in 2. Transthoracic Ivor-Lewis esophagectomy with laparoscopy and thoracoscopy was performed in 28, a 3-stage esophagectomy in 3, and transhaital esophagectomy in 1. There was no operative mortality and median hospital stay was 10.5 days for patients treated with minimally invasive esophagectomy. This compares with an operative mortality of 8.9% and median hospital stay of 17 days for open esophagectomy in our institution. PMID- 21640920 TI - Cancer immunotherapy. AB - Immune-based therapies for cancer are now commonplace. Cytokine therapy, including interferon and interleukin-2, is safe in the community setting. The US Food and Drug Administration has recently approved sipuleucel-T for the treatment of advanced prostate cancer, the first therapeutic cancer vaccine to meet this level of efficacy. The therapeutic use of monoclonal antibodies directed against proteins controlling various cell functions, including growth and modulation of immune response, has become so pervasive that the oncologist, whether surgeon or medical oncologist, must be familiar with indications, contraindications, and the associated toxicities. PMID- 21640921 TI - Breast cancer care in the community: challenges, opportunities, and outcomes. AB - In the USA, 80% of patients with breast cancer are treated by community breast surgeons. NCDB data indicate that there are only small differences in outcomes between lower volume cancer programs and higher volume programs. There is some evidence that breast cancer patients of high-volume breast focused surgeons may have improved outcomes. This article discusses the challenges community breast surgeons face and some ways that the quality of care could be monitored and improved. Quality reporting programs of the Commission on Cancer and Mastery of Breast Surgery Program of the American Society of Breast Surgeons are recommended as tools to track and improve outcomes in breast cancer care. PMID- 21640922 TI - The National Accreditation Program for Breast Centers: quality improvement through standard setting. AB - The breast center concept developed in response to a fragmented inefficient system to evaluate and manage patients with diseases of the breast. The National Accreditation Program for Breast Centers (NAPBC) accreditation is granted only to those centers that have voluntarily committed to provide the best in the diagnosis and treatment of breast cancer and are able to comply with the established NAPBC standards. Each center must undergo a rigorous evaluation and review of its performance and compliance with the NAPBC standards. The NAPBC is in the process of defining the most efficient methods of data collection for the NAPBC-accredited programs. PMID- 21640923 TI - Improving uniformity of care for colorectal cancers through National Quality Forum quality indicators at a Commission on Cancer-accredited community based teaching hospital. AB - The National Cancer Data Base (NCDB) provides feedback on adherence to National Quality Forum (NQF)-endorsed measures to promote best outcomes in colorectal cancer. We examined the care delivered to patients with colorectal cancer at our institution and developed a protocol to enhance nodal retrieval and to ensure that patients with fewer than 12 nodes are considered for adjuvant chemotherapy. Few patients met the NQF criteria for adjuvant radiation. A protocol was developed to address this issue, and this provides a model for use in a multidisciplinary effort to improve adherence to measures associated with best outcomes in colorectal cancer. PMID- 21640924 TI - Proceedings of the Falk Benelux IBD summit. Preface. PMID- 21640925 TI - Novel developments in IBD-related sclerosing cholangitis. AB - Primary sclerosing cholangitis is often regarded as an autoimmune disorder and occurs frequently in relation to inflammatory bowel disease. The ongoing fibro obliterative process of the biliary tree ensues in liver failure or cholangiocarcinoma in 12-18 years. PSC patients with concurrent IBD are at increased risk of developing colorectal carcinoma. Ursodeoxycholic acid, which is widely prescribed in PSC, is despite intensive clinical research still not proven to halt disease progression. Nor-ursodeoxycholic acid seems promising in animal models of cholestasis. Novel compounds that are involved in the immunological axis between the gut and the liver await clinical testing in PSC. PMID- 21640926 TI - Novel development in extraintestinal manifestations and spondylarthropathy. AB - The important co-existence of spondylarthritis (SpA) and inflammatory bowel disease (IBD) within the same individual suggests common etiopathogenic mechanisms. This is supported by intriguing similarities between both diseases at the subclinical and molecular level. The recent advances in IBD genetics have led to the identification of common pathways involved in both IBD and SpA, including bacterial recognition and ER stress. This offers the opportunity to develop potential new therapeutic strategies for both diseases. Transgenic animals which develop both joint and gut inflammation (like the TNF(DeltaARE) mice and the HLA B27 transgenic rats) are a very useful tool to test such novel therapeutics and to get further mechanistic insight into the pathogenetic link between SpA and IBD. This review will focus on the recent scientific progress in our understanding of the link between SpA and IBD. Based on this, potential novel therapeutic strategies are discussed. PMID- 21640927 TI - Necessity of phenotypic classification of inflammatory bowel disease. AB - Inflammatory bowel diseases (IBD) are classically divided in Crohn's disease (CD) and ulcerative colitis (UC). However, these two entities are still heterogeneous and a further classification in subphenotypes is necessary. Clinical subphenotypes are easy to use, do not necessitate complicated tests and can already give very important information for the management of the patients. In CD, clinical subphenotypes are based on age at diagnosis, disease location and disease behaviour. Age at diagnosis allows to differentiating paediatric CD, classical young adult onset and more seldom CD of the elderly. These categories are associated with a different risk of development of complications and disabling disease and may have partly different pathophysiology. The classification on disease behaviour, including stricturin, penetrating or uncomplicated disease may have an impact on reponse to medical treatment and need for surgery. Finally the classification based on location is particularly relevant since it has been associated with different types of complications. Particularly ileal disease has been associated with the risk of surgery and colonic (particularly rectal) disease, with the risk of perianal disease. In UC, the classification in subphenotypes is essentially based on disease location, distinguishing proctitis, left-sided colitis and extensive colitis. This subclassification also has a very significant clinical relevance since extensive colitis has been associated with and increased risk of colon cancer, colectomy and even in some studies, mortality. PMID- 21640928 TI - IBD-related carcinoma and lymphoma. AB - Patients with inflammatory bowel disease (IBD) have an increased risk of developing colorectal cancer (CRC). Risk factors are extent and severity of colonic inflammation, concurrent primary sclerosing cholangitis, and a positive family history of sporadic CRC. The chromosomal instability, microsatellite instability and hypermethylation pathways form the molecular background of IBD related carcinogenesis, which is not different from sporadic CRC. The dysplasia carcinoma sequence of IBD-related colorectal carcinogenesis makes patients suitable for endoscopic surveillance. In the future, new molecular biomarkers and endoscopic techniques may improve early detection of precursor lesions of IBD related CRC. The potential of aminosalicylates and ursodeoxycholic acid as chemopreventive agents needs to be studied in randomized clinical trials. Patients with IBD who are being treated with thiopurines have a slightly increased risk of developing lymphoproliferative disorders, whereas patients with small bowel Crohn's disease have a high relative risk and a small absolute risk of developing small bowel adenocarcinoma. PMID- 21640929 TI - To what extent are genetics clinically useful? AB - The volume of research undertaken on the genetic susceptibility of inflammatory bowel diseases has been tremendous. International collaborative efforts which initiated in 1997 and have not stopped since, led to the identification at present of more than 100 IBD risk loci. Yet, only 25% of the genetic variance is explained. It is hypothesized that rare variants, other forms of genetic variation (copy number variation), as well as gene-gene and geneenvironment interactions, explain the missing heritability. From all genes identified, the pattern recognition receptor NOD2/CARD15 is still the most understood at present. The field of IBD genetics has translated itself so far in identifying pathways important for disease pathogenesis (autophagy, Th17/IL23, pattern recognition receptors and innate immunity, barrier integrity), some of which have promising therapeutic consequences. Second, although genetic testing will most likely have no or a very limited role in diagnosing patients, it is anticipated that genetic markers will be implemented in an integrated prognostic approach. Efforts to predict disease course and response to therapy have shown interesting results. PMID- 21640932 TI - Does the mitral valve recoil after percutaneous balloon valvotomy? AB - AIM: The significance of passive stretching of the mitral valve as a contributor to valve opening, after percutaneous transvenous mitral commissurotomy (PTMC), is not known. Our objective was to determine whether any acute reduction in valve area occurs due to recoil of stretched valve structures. METHODS AND RESULTS: In a prospective observational study, we evaluated nine patients (age 30.1+/-8.0 years; median valve score 7) who underwent PTMC. We calculated mitral valve area (MVA) before, immediately after, and at 10 and 30 min after valvotomy. There was no acute reduction in MVA after successful PTMC. But there was a significant increase in MVA at 30 min, from that measured immediately after the procedure (1.8+/-0.4 to 2.0+/-0.4 cm(2); P=.048). This was attributable to the continuing fall in pulmonary artery wedge (PAW) pressures (17+/-3 to 15+/-3 mmHg; P=.003) and transmitral gradients (8+/-3 to 7+/-2 mmHg; P=.037). CONCLUSION: Passive stretching of the valve apparatus does not play an important role in valve opening after PTMC in young patients with favorable valve morphology. PMID- 21640931 TI - Coronary stenting with MGuard: extended follow-up of first human trial. AB - OBJECTIVES: To evaluate feasibility and safety of MGuard based percutaneous coronary interventions (PCI) in vein grafts (VG) and native coronaries (NC). BACKGROUND: Distal embolization is a frequent complication of PCI of VG and NC during acute coronary syndromes (ACS). MGuard was a stent designed to reduce embolization. METHODS: Prospective, single arm, two-center trial assessing the feasibility and safety of MGuard-based PCI with post-PCI clinical and laboratory monitoring including: cardiac biomarkers, ECG and 6-month angiography. RESULTS: Forty-one patients with mean age of 68.2+/-10.1 years were enrolled. Mean VG age (n=23) was 14.4+/-4.3 years. All patients received heparin, clopidogrel and aspirin; while none received glycoprotein IIb/IIIa inhibitors, or embolic protection device (EPD). Device and procedural success were 100% and 95.1% respectively. Two patients (4.9%) experienced procedure-related creatinine phosphokinase rise. At 6 months one patient had myocardial infarction and 19.5% had target vessel revascularization (TLR). Late follow up (12-27 months) revealed one additional TLR. CONCLUSION: MGuard based PCI of NC and VG appears encouraging especially in view of unfavorable patient and lesion characteristics. Efficacy needs to be further established in larger randomized trials. PMID- 21640933 TI - Long-term clinical outcomes after intravascular brachytherapy for instent restenosis and de novo coronary artery lesions in percutaneous coronary intervention. AB - BACKGROUND: We examined the long term clinical outcomes after administration of intravascular brachytherapy (IVBT) for instent restenosis (ISR) and de novo coronary artery lesions in percutaneous coronary intervention (PCI). METHODS AND MATERIALS: From May 2000 to August 2004, 129 symptomatic patients with ISR and de novo coronary artery lesions were treated with intravascular beta radiation after successful PCI. The primary end-point was major adverse cardiac event (MACE), i.e., a composite of all-cause death, myocardial infarction and target lesion revascularization (TLR) within 5 years of follow-up. RESULTS: The mean age of patients was 64+10 years with a male predominance (78%). The majority of patients had diffuse bare metal instent restenotic lesions and 19 patients (15%) had de novo coronary artery lesions. From hospital discharge to follow-up at 5 years (mean follow-up period 75.3+17.7 months), the annual consecutive MACE rates were 16.3%, 13.4%, 8%, 12.2% and 6.6% respectively and were mainly driven by the need for TLR. Left anterior descending artery (LAD) as target vessel of PCI was an independent predictor of long term MACE (OR: 3.5; 95% confidence interval:1.2 10.6; P=.03). There were six cases of stent thrombosis (cumulative incidence of 4.7%) with case fatality rate of 33% (2/6). CONCLUSION: MACE rates remained high post IVBT at 5 years of follow-up and were mainly driven by the need for TLR. LAD as target vessel of PCI was an independent predictor of long term MACE. PMID- 21640934 TI - Percutaneous patent foramen ovale closure: outcomes with the Premere and Amplatzer devices. AB - BACKGROUND: Transcatheter closure of patent foramen ovale (PFO) has rapidly evolved as the preferred management strategy for the prevention of recurrent cerebrovascular events in patients with cryptogenic stroke and presumed paradoxical embolus. There is limited outcome data in patients treated with this therapy particularly for the newer devices. METHODS: Data from medical records, catheter, and echocardiography databases on 70 PFO procedures performed was collected prospectively. RESULTS: The cohort consisted of 70 patients (mean age 43.6 years, range 19 to 77 years), of whom 51% were male. The indications for closure were cryptogenic cerebrovascular accident (CVA) or transient ischemic attack (TIA) in 64 (91%) and peripheral emboli in two (2.8%) patients and cryptogenic ST-elevation myocardial infarction in one (1.4%), refractory migraine in one (1.4%), decompression sickness in one (1.4%), and orthodeoxia in one (1.4%) patient, respectively. All patients had demonstrated right-to-left shunting on bubble study. The procedures were guided by intracardiac echocardiography in 53%, transesophageal echocardiography in 39%, and the remainder by transthoracic echo alone. Devices used were the Amplatzer PFO Occluder (AGA Medical) (sizes 18-35 mm) in 49 (70%) and the Premere device (St. Jude Medical) in 21 (30%). In-hospital complications consisted of one significant groin hematoma with skin infection. Echocardiographic follow-up at 6 months revealed that most patients had no or trivial residual shunt (98.6%), while one patient (1.4%) had a mild residual shunt. At a median of 11 months' follow-up (range 1 month to 4.3 years), no patients (0%) experienced further CVA/TIAs or paradoxical embolic events during follow-up. CONCLUSION: PFO causing presumed paradoxical embolism can be closed percutaneously with a low rate of significant residual shunting and very few complications. Recurrent index events are uncommon at medium-term (up to 4 years) follow-up. PMID- 21640935 TI - Contemporary outcomes of percutaneous intervention in chronic total coronary occlusions due to in-stent restenosis. AB - BACKGROUND: Limited data exist on the treatment of chronic total occlusions (CTO) due to in-stent restenosis (ISR). METHODS: We reviewed the procedural techniques and outcomes of 21 consecutive interventions in CTOs due to ISR. RESULTS: Mean age was 60+/-8 years, and all patients were men. The target lesion was located in the right coronary artery in 38%, left anterior descending or diagonal in 48%, or circumflex/obtuse marginal in 14%. One CTO lesion was treated in each patient. Two patients (10%) had prior unsuccessful attempt for CTO intervention and 14% had prior coronary artery bypass graft surgery. The average CTO age was 6.3+/-4.6 years. The overall ISR CTO procedural success rate was 71%. Procedural failure was due to inability to cross the CTO lesion in all unsuccessful cases. Failure to cross in CTOs located in the left anterior descending artery was due to the presence of a large side branch proximal to the CTO, whereas in the right coronary artery it was due to tortuosity. Retrograde interventions were attempted in four patients and were successful in one. CONCLUSION: Success rates for ISR CTO interventions remain relatively low due to failure to cross the lesion. Several factors, such as long occlusion time, tortuosity, and presence of a large side branch proximal to the CTO may be associated with ISR CTO crossing failure. PMID- 21640936 TI - Ascending aortic pseudoaneurysm, a ticking bomb after cardiac surgery. AB - Ascending aortic pseudoaneurysm (AAP) is a rare but serious complication after cardiothoracic surgery. Patients typically present with chest pain, fever, or mass effects, but asymptomatic individuals with abnormal imaging results have been reported. Currently, there are no established guidelines regarding the management of AAP, but most authors recommend surgical treatment even in asymptomatic patients. Here, we describe a 39-year-old woman with Marfan syndrome who developed an AAP approximately 7 years after cardiac surgery. She was treated conservatively and remained asymptomatic until the past 2 years when she developed progressive chest pain and dyspnea upon exertion. Computed tomography angiography, coronary angiography, and ascending aortography were performed and revealed the presence of a pseudoaneurysm of the ascending aorta. Surgical intervention was successful and she was discharged home without significant issues. PMID- 21640937 TI - The value of using fractional flow reserve measurements in helping to diagnose acute left ventricular failure in the presence of normal left ventricular systolic function. PMID- 21640938 TI - Infective endocarditis caused by Moraxella nonliquefaciens in a percutaneous aortic valve replacement. AB - First case of infective endocarditis on a percutaneous aortic valve replacement due to Moraxalla nonliquefaciens in 64 year old woman. It was successfully treated medically with antibiotics. She had not suitable for surgical aortic valve replacement due to 3 sternotomies for thymoma resection and subsequent radiotherapy with blocked major thoracic veins. Due to her azathioprine immunosuppresion (myasthenia) she may have been at increased endocarditis risk. We suggest prophylactic antibiotics at implant for this group in future. PMID- 21640939 TI - Sterile radial artery granuloma after transradial cardiac catheterization. AB - Transradial cardiac catheterization has lower rates of arterial access site complications than transfemoral procedures. However, there are complications that are unique to the transradial route. We present the case of a sterile granuloma occurring at the site of radial arterial access as a reaction to the hydrophilic coating on the sheath. The clinical presentation was suggestive of an infected pseudoaneurysm. Awareness of this entity may help clinicians avoid unnecessary surgical procedures, as these granulomata are transient self-limiting reactions. PMID- 21640941 TI - Biodiversity in face of human activities. Foreword. PMID- 21640942 TI - Why should we be concerned about loss of biodiversity. AB - Over the past century, documented extinctions in well-studied taxonomic groups have been at rates one hundred-fold to one thousand-fold above the average extinction rates seen over the half billion year sweep of the fossil record. But for most groups, particularly invertebrates, we are very uncertain how many species there are on Earth today, much less rates of extinction. PMID- 21640943 TI - Biodiversity is not (and never has been) a bed of roses! AB - Over the last decades, the critical study of fossil diversity has led to significant advances in the knowledge of global macroevolutionary patterns of biodiversity. The deep-time history of life on Earth results from background originations and extinctions defining a steady-state, nonstationary equilibrium occasionally perturbed by biotic crises and "explosive" diversifications. More recently, a macroecological approach to the large-scale distribution of extant biodiversity offered new, stimulating perspectives on old theoretical questions and current practical problems in conservation biology. However, time and space are practically distinct, but functionally related dimensions of ecological systems. This calls for a spatially-integrated study of biodiversity dynamics at an evolutionary timescale. Indeed, the biosphere is a complex adaptive system whose study cannot be arbitrarily reduced to any single spatial- and/or temporal scale level of resolution without a loss of content. From such an integrated perspective, a simple fact emerges: in a physically heterogeneous and ever changing world, spatiotemporal variations in biodiversity are the rule-not the exception. PMID- 21640944 TI - The impact of global change on terrestrial Vertebrates. AB - Examples of the impact of human activities on Vertebrate populations abound, with famous cases of extinction. This article reviews how and why Vertebrates are affected by the various components of global change. The effect of direct exploitation, while strong, is currently superseded by changes in use of all sorts, while climate change has started having significant effects on some Vertebrate populations. The low maximum growth rate of Vertebrate populations makes them particularly sensitive to global change, while they contribute relatively modestly to major ecosystem services. One may conclude that unless they are considered as sentinels of the biological consequences of global changes, their situation will go on strongly deteriorating, in particular under the influence of interactions of different components of global change such as changes in use and climate change. PMID- 21640945 TI - Scientific contributions of extensive biodiversity monitoring. AB - To develop a complete and informative biodiversity observation system, it is necessary to compare the strengths and limits of various monitoring schemes. In this article, we examine the various advantages of extensively monitoring fine grained spatial variations of biodiversity, where the prominent traits of many species within a community (abundance, phenology, etc.) are regularly recorded at numerous sites over a large territory, usually via human observation networks. Linking these variations with environmental factors sheds lights on the major mechanisms leading to changes in biodiversity, thus increasing our knowledge of macroecology and community ecology. This extensive monitoring allows us to assess diffuse effects, contributing to the sound use of the precautionary principle. Combined with site-focused monitoring, information gathered from extensive monitoring provides the raw material necessary to build biodiversity scenarios. PMID- 21640946 TI - An ethical issue in biodiversity science: The monitoring of penguins with flipper bands. AB - Individual marking is essential to study the life-history traits of animals and to track them in all kinds of ecological, behavioural or physiological studies. Unlike other birds, penguins cannot be banded on their legs due to their leg joint anatomy and a band is instead fixed around a flipper. However, there is now detailed evidence that flipper-banding has a detrimental impact on individuals. It can severely injure flipper tissues, and the drag effect of their flipper bands results in a higher energy expenditure when birds are moving through the water. It also results in lower efficiency in foraging, since they require longer foraging trips, as well as in lower survival and lower breeding success. Moreover, due to the uncertainty of the rate of band loss, flipper bands induce a scientific bias. These problems, which obviously have serious ethical implications, can be avoided with alternative methods such as radiofrequency identification techniques. PMID- 21640947 TI - Ecosystem dynamics, biological diversity and emerging infectious diseases. AB - In this article, we summarize the major scientific developments of the last decade on the transmission of infectious agents in multi-host systems. Almost sixty percent of the pathogens that have emerged in humans during the last 30-40 years are of animal origin and about sixty percent of them show an important variety of host species besides humans (3 or more possible host species). In this review, we focus on zoonotic infections with vector-borne transmission and dissect the contrasting effects that a multiplicity of host reservoirs and vectors can have on their disease dynamics. We discuss the effects exerted by host and vector species richness and composition on pathogen prevalence (i.e., reduction, including the dilution effect, or amplification). We emphasize that, in multiple host systems and for vector-borne zoonotic pathogens, host reservoir species and vector species can exert contrasting effect locally. The outcome on disease dynamics (reduced pathogen prevalence in vectors when the host reservoir species is rich and increased pathogen prevalence when the vector species richness increases) may be highly heterogeneous in both space and time. We then ask briefly how a shift towards a more systemic perspective in the study of emerging infectious diseases, which are driven by a multiplicity of hosts, may stimulate further research developments. Finally, we propose some research avenues that take better into account the multi-host species reality in the transmission of the most important emerging infectious diseases, and, particularly, suggest, as a possible orientation, the careful assessment of the life-history characteristics of hosts and vectors in a community ecology-based perspective. PMID- 21640948 TI - Functional diversity of terrestrial microbial decomposers and their substrates. AB - The relationship between biodiversity and biogeochemical processes gained much interest in light of the rapidly decreasing biodiversity worldwide. In this article, we discuss the current status, challenges and prospects of functional concepts to plant litter diversity and microbial decomposer diversity. We also evaluate whether these concepts permit a better understanding of how biodiversity is linked to litter decomposition as a key ecosystem process influencing carbon and nutrient cycles. Based on a literature survey, we show that plant litter and microbial diversity matters for decomposition, but that considering numbers of taxonomic units appears overall as little relevant and less useful than functional diversity. However, despite easily available functional litter traits and the well-established theoretical framework for functional litter diversity, the impact of functional litter diversity on decomposition is not yet well enough explored. Defining functional diversity of microorganisms remains one of the biggest challenges for functional approaches to microbial diversity. Recent developments in microarray and metagenomics technology offer promising possibilities in the assessment of the functional structure of microbial communities. This might allow significant progress in measuring functional microbial diversity and ultimately in our ability to predict consequences of biodiversity loss in the decomposer system for biogeochemical processes. PMID- 21640949 TI - Soil microbial diversity: Methodological strategy, spatial overview and functional interest. AB - Since the development of industrialization, urbanization and agriculture, soils have been subjected to numerous variations in environmental conditions, which have resulted in modifications of the taxonomic diversity and functioning of the indigenous microbial communities. As a consequence, the functional significance of these losses/modifications of biodiversity, in terms of the capacity of ecosystems to maintain the functions and services on which humanity depends, is now of pivotal importance. In this context, one of the main challenges in soil microbial ecology is to better understand and predict the processes that drive soil microbial diversity and the link between diversity and ecosystem process. This review describes past, present and ongoing conceptual and methodological strategies employed to better assess and understand the distribution and evolution of soil microbial diversity with the aim of increasing our capacity to translate such diversity into soil biological functioning and, more widely, into ecosystem services. PMID- 21640950 TI - Ecological solidarity as a conceptual tool for rethinking ecological and social interdependence in conservation policy for protected areas and their surrounding landscape. AB - Policy for biodiversity conservation must evolve to cope with the increasing human footprint on natural systems. A major issue here is the need for policy for protected areas, which integrates their surrounding landscape and local human populations in the construction of socially grounded measures. To illustrate current conceptual thinking in this direction we present and provide a conceptual basis for a recent initiative in national park policy in France that is based on "ecological solidarity". In the light of other policy ideas and tools that have recently emerged for the co-construction of conservation policy, we argue that this concept provides an imaginative step towards consolidating ecological and social interdependence in biodiversity policy that goes beyond statutory park boundaries. PMID- 21640951 TI - River networks as biodiversity hotlines. AB - For several years, measures to insure healthy river functions and to protect biodiversity have focused on management at the scale of drainage basins. Indeed, rivers bear witness to the health of their drainage basins, which justifies integrated basin management. However, this vision should not mask two other aspects of the protection of aquatic and riparian biodiversity as well as services provided by rivers. First, although largely depending on the ecological properties of the surrounding terrestrial environment, rivers are ecological systems by themselves, characterized by their linearity: they are organized in connected networks, complex and ever changing, open to the sea. Second, the structure and functions of river networks respond to manipulations of their hydrology, and are particularly vulnerable to climatic variations. Whatever the scale considered, river networks represent "hotlines" for sharing water between ecological and societal systems, as well as for preserving both systems in the face of global change. River hotlines are characterized by spatial as well as temporal legacies: every human impact to a river network may be transmitted far downstream from its point of origin, and may produce effects only after a more or less prolonged latency period. Here, I review some of the current issues of river ecology in light of the linear character of river networks. PMID- 21640952 TI - Marine biodiversity characteristics. AB - Oceans contain the largest living volume of the "blue" planet, inhabited by approximately 235-250,000 described species, all groups included. They only represent some 13% of the known species on the Earth, but the marine biomasses are really huge. Marine phytoplankton alone represents half the production of organic matter on Earth while marine bacteria represent more than 10%. Life first appeared in the oceans more than 3.8 billion years ago and several determining events took place that changed the course of life, ranging from the development of the cell nucleus to sexual reproduction going through multi-cellular organisms and the capture of organelles. Of the 31 animal phyla currently listed, 12 are exclusively marine phyla and have never left the ocean. An interesting question is to try to understand why there are so few marine species versus land species? This pattern of distribution seems pretty recent in the course of Evolution. From an exclusively marine world, since the beginning until 440 million years ago, land number of species much increased 110 million years ago. Specific diversity and ancestral roles, in addition to organizational models and original behaviors, have made marine organisms excellent reservoirs for identifying and extracting molecules (>15,000 today) with pharmacological potential. They also make particularly relevant models for both fundamental and applied research. Some marine models have been the source of essential discoveries in life sciences. From this diversity, the ocean provides humankind with renewable resources, which are highly threatened today and need more adequate management to preserve ocean habitats, stocks and biodiversity. PMID- 21640953 TI - The diversity of the ecosystem services concept and its implications for their assessment and management. AB - The ecosystem services concept is used in different scientific disciplines and is spreading into policy and business circles to draw attention to the benefits that people receive from biodiversity and ecosystems. However, the concept remains multiform and is used interchangeably with a range of other terms such as ecological, landscape or environmental services. We argue that lexical differences, in fact, result from different understandings of the concept, which could slow its use in nature conservation or sustainable resource use. An application to semi-natural grasslands shows that such differences could lead to very different assessments, of quality, quantity and location of ecosystem services. We argue that a compromise must be found between a broad and simple definition, which is useful for communicating the concept and large-scale policies, and a more refined definition for research and implementation goals such as environmental management and national and international assessments and accounting. PMID- 21640954 TI - Biodiversity, evolution and adaptation of cultivated crops. AB - The human diet depends on very few crops. Current diversity in these crops is the result of a long interaction between farmers and cultivated plants, and their environment. Man largely shaped crop biodiversity from the domestication period 12,000 B.P. to the development of improved varieties during the last century. We illustrate this process through a detailed analysis of the domestication and early diffusion of maize. In smallholder agricultural systems, farmers still have a major impact on crop diversity today. We review several examples of the major impact of man on current diversity. Finally, biodiversity is considered to be an asset for adaptation to current environmental changes. We describe the evolution of pearl millet in West Africa, where average rainfall has decreased over the last forty years. Diversity in cultivated varieties has certainly helped this crop to adapt to climate variation. PMID- 21640955 TI - Dynamic management of crop diversity: From an experimental approach to on-farm conservation. AB - In agricultural systems, biodiversity includes diversity within species and among species and provides many benefits for production, resilience and conservation. This article addresses the effects of a strategy of in situ conservation called dynamic management (DM) on population evolution, adaptation and diversity. Two French DM initiatives are considered, the first one corresponding to an experimental context, the second to an on-farm management. Results from a study over 26 years of experimental DM of bread wheat (Triticum aestivum L.) are first presented, including the evolution of agronomic traits and genetic diversity at neutral and fitness related loci. While this experiment greatly increased scientific knowledge of the effects of natural selection on cultivated populations, it also showed that population conservation cannot rely only on a network of experimental stations. In collaboration with a farmers' network in France, researchers have begun studying the effects of on-farm DM (conservation and selection) on diversity and adaptation. Results from these studies show that on-farm DM is a key element for the long-term conservation and use of agricultural biodiversity. This method of in situ conservation deserves more attention in industrialised countries. PMID- 21640956 TI - Valuing biodiversity and ecosystem services: Why put economic values on Nature? AB - The evaluation of ecosystems and biodiversity has become an important field of investigation for economists. Although their interest has been largely motivated by the search for arguments in favour of broader conservation policies, both the methods and the meaning of the results remain controversial. This article aims at clarifying the interest and limitations of these works, by revisiting a number of issues, such as the economic qualification of the services that human societies take from biodiversity and ecological systems in general, the specificities of their contribution to human well-being and the consequences of a valuation of biodiversity based on ecosystem services. We conclude with a discussion of the purposes of evaluations: improving public policies or creating new markets? PMID- 21640957 TI - 2010: A new beginning for biodiversity? AB - Proclaimed "International Year of Biodiversity", will 2010 hold all its promises? Reminder: initiated by the Convention on Biological Diversity ratified after the global summit in Rio de Janeiro, delegations from more than one hundred countries gathered in Johannesburg in 2002 and committed themselves to slowing the erosion of biodiversity by 2010. The European Union was more ambitious (or reckless?) and even spoke about halting this erosion (European Environment Agency, Progress towards the European 2010 biodiversity target, 2009)! Well, that date has come and the overall appraisal that has been made formally in Nagoya in October this year was not so brilliant (see Leadley et al., 2010)-but the same slogan has been launched for 2020! The aim here is not to repeat that appraisal, but, after considering the broad outlines, to evoke some of the issues and challenges that inevitably result from the great question of the protection and management of global biodiversity. PMID- 21640958 TI - Feeling-of-knowing for songs and instrumental music. AB - We explored the differences between metamemory judgments for titles as well as for melodies of instrumental music and those for songs with lyrics. Participants were given melody or title cues and asked to provide the corresponding titles or melodies or feeling of knowing (FOK) ratings. FOK ratings were higher but less accurate for titles with melody cues than vice versa, but only in instrumental music, replicating previous findings. In a series of seven experiments, we ruled out style, instrumentation, and strategy differences as explanations for this asymmetry. A mediating role of lyrics between the title and the melody in songs was also ruled out. What emerged as the main explanation was the degree of familiarity with the musical pieces, which was manipulated either episodically or semantically, and within this context, lyrics appeared to serve as an additional source of familiarity. Results are discussed using the Interactive Theory of how FOK judgments are made. PMID- 21640959 TI - [Sarcoid granulomas in facial cosmetic filler material: induction by interferon alpha and ribavirin in a patient with hepatitis C]. PMID- 21640960 TI - [Sentinel node biopsy in melanoma: an update]. PMID- 21640961 TI - [Myeloid sarcoma in the area of a skin flap]. PMID- 21640962 TI - [Epidermal effacement in malignant melanoma]. PMID- 21640963 TI - Exenteration as a primary treatment for locally advanced cervical cancer: long term results and prognostic factors. AB - OBJECTIVE: Whereas pelvic exenteration is an established therapy for the treatment of recurrent cervical carcinoma, it is not often performed for primary locally advanced cervical cancer (LACC). STUDY DESIGN: This retrospective study analyzed survival data and prognostic factors of 35 patients with LACC who were treated by pelvic exenteration. RESULTS: After surgery, 33 patients (97%) were macroscopically free of tumor. In 20 patients, pelvic lymph nodes (LN) were involved, and in 6 of these, metastatic tissue had reached the paraaortal nodes. Overall, the patients' mean 5 year survival was 43%, and the median survival time was 30 months; these values ranged from 15% to 70% and from 15 to 44 months, respectively, depending on LN involvement (P=.006). Pelvic LN involvement was the only significant factor for overall survival found in the multivariate analysis (P=.02). CONCLUSION: In LACC with free LNs and no distant metastases, pelvic exenteration has good long-term results. PMID- 21640964 TI - Evaluation of transobturator tension-free vaginal tapes in the management of women with mixed urinary incontinence: one-year outcomes. AB - OBJECTIVE: To assess the efficacy of transobturator tapes in the management of women with urodynamic mixed urinary incontinence (UI). STUDY DESIGN: A secondary analysis of a prospective randomized study; 83 women with mixed UI on urodynamics and predominant stress UI symptoms were recruited and randomly assigned to undergo "outside-in" or inside-out transobturator tapes. Preoperative assessment included urodynamic assessment and completion of validated symptom severity and quality of life (QoL) questionnaires. The primary outcome was patient-reported success rates at 1-year as assessed by the Patient Global Impression of Improvement (very much/much improved). Secondary outcomes included changes in preoperative urgency/urgency incontinence, changes in QoL scores and comparison between the 2 types of transobturator tapes. RESULTS: Seventy-seven women completed 1-year follow-up (outside-in [n=42] vs inside-out [n=35]). The patient reported success rate and objective cure rate were 75% and 90%, respectively. At 1-year follow-up; 40 women (52%) and 31 women (57.4%) reported cure in their preoperative urgency and urgency incontinence, respectively. A total of 74% reported>=10 point improvement in QoL scores. CONCLUSION: In women with urodynamic mixed incontinence and predominant stress UI, transobturator tapes were associated with good patient-reported success rate at 1 year. Urgency/ urgency incontinence are cured in over 50% of women. PMID- 21640965 TI - Maternal treatment with opioid analgesics and risk for birth defects: additional considerations. PMID- 21640966 TI - Insulation failure in robotic and laparoscopic instrumentation: a prospective evaluation. AB - OBJECTIVE: The purpose of this study was to detect the incidence, prevalence, and location of insulation failures (IFs) in laparoscopic and robotic instruments. STUDY DESIGN: In phase A, a total of 78 robotic and 298 laparoscopic instruments were tested at 20 W and 2.64 kV at Mayo Clinic in Arizona. In phase B, 60 robotic and 308 laparoscopic instruments were tested at 20 W/1 kV and 20 W/4.2 kV, respectively. RESULTS: In phase A, the robotic group showed a higher prevalence (25/78; 32%) and incidence of IFs after 10 uses (35/44 instruments; 80%) when compared with laparoscopy (prevalence, 39/298 [13%]; incidence, 68/189 [36%]; P<.05). In phase B, IFs were detected in 81.7% of the robotic instruments and in 19.5% of the laparoscopic instruments (P<.005). CONCLUSION: There is a high incidence and prevalence of IF in endoscopic instrumentation that is more common in the robotic group. PMID- 21640967 TI - Human amnion epithelial cells as a treatment for inflammation-induced fetal lung injury in sheep. AB - OBJECTIVE: The purpose of this study was to determine whether human amnion epithelial cells (hAECs) can modulate the pulmonary developmental consequences of intrauterine inflammation in fetal sheep that are exposed to intraamniotic lipopolysaccharide (LPS) injection. STUDY DESIGN: At 117 days' gestation, fetal sheep (n=16) received intraamniotic LPS (20 mg). hAECs were delivered at 0, 6, and 12 hours into the fetal jugular vein (n=4), trachea (n=4), or both (n=4). Controls (n=6) received equivalent administration of saline solution. Lungs were collected at 124 days. RESULTS: Intraamniotic LPS caused pulmonary inflammation and altered lung structure and function. hAECs attenuated changes in lung function and structure that had been induced by LPS: lung volume, 40 cm H2O (P<.05, intravenous+intratracheal hAECs vs LPS), tissue-to-airspace ratio (P<.05, intravenous+intratracheal hAECs vs LPS), and septal crest density (P<.001, all hAEC groups vs LPS). Leukocyte infiltration of the lungs was not reduced by hAECs; however, inflammatory cytokines were reduced (tumor necrosis factor-alpha, P<.01, vs LPS; interleukin-1b, P<.01, vs LPS; interleukin-6, P<.01 vs LPS). Surfactant protein A and C messenger RNA was increased by LPS, although this was not statistically significant (P>.05 vs control); there were significant increases in all hAEC-treated animals (surfactant protein-A, P<.05 vs LPS; surfactant protein-C, P<.01 vs LPS). CONCLUSION: Human amnion epithelial cells attenuate the fetal pulmonary inflammatory response to experimental intrauterine inflammation and reduce, but (as administered in our study) do not prevent, consequent alterations in lung development. PMID- 21640968 TI - Systematic review of first-trimester vitamin D normative levels and outcomes of pregnancy. AB - OBJECTIVE: We undertook a systematic review to assess normative levels of vitamin D in early pregnancy and association with subsequent pregnancy outcomes. STUDY DESIGN: Medline and Embase databases and reference lists were searched. Inclusion criteria were pregnant populations, blood sample taken during the first trimester, and serum hydroxyvitamin D levels assessed. RESULTS: Eighteen studies reported vitamin D levels in first trimester (n = 11-3730), and 5 examined pregnancy outcomes. Mean vitamin D concentrations differed when stratified by ethnicity: white (mean [SD]: 29.4 [11.7] to 73.1 [27.1] nmol/L) and nonwhite (15.2 [12.1] to 43 [12] nmol/L). Most studies used general population cut points to define deficiency and found a large proportion of women deficient. Two articles examined risk of preeclampsia and reported differing findings, whereas 2 of 3 found low levels associated with increased risk of small-for-gestational age births. CONCLUSION: There is no clear definition of vitamin D deficiency in pregnancy and insufficient evidence to suggest low vitamin D levels in early pregnancy are associated with adverse pregnancy outcomes. PMID- 21640969 TI - Management of women treated with buprenorphine during pregnancy. AB - The management of pregnancy and delivery of a woman on opiate-substitution therapy with buprenorphine requires a coordinated team approach by social services, addiction medicine, obstetrics, and pediatrics. Her obstetrical care is further complicated by the unique pharmacology of buprenorphine and the issues of pain management. Obstetrical providers should be familiar with the complex issues surrounding the optimal care of these women. PMID- 21640970 TI - Effective physician-nurse communication: a patient safety essential for labor and delivery. AB - Effective communication is a hallmark of safe patient care. Challenges to effective interprofessional communication in maternity care include differing professional perspectives on clinical management, steep hierarchies, and lack of administrative support for change. We review principles of high reliability as they apply to communication in clinical care and discuss principles of effective communication and conflict management in maternity care. Effective clinical communication is respectful, clear, direct, and explicit. We use a clinical scenario to illustrate an historic style of nurse-physician communication and demonstrate how communication can be improved to promote trust and patient safety. Consistent execution of successful communication requires excellent listening skills, superb administrative support, and collective commitment to move past traditional hierarchy and professional stereotyping. PMID- 21640971 TI - Discoidin domain receptor 1 is a major mediator of inflammation and fibrosis in obstructive nephropathy. AB - The interactions between tubulointerstitial infiltrating cells and the extracellular matrix play an important role in regulating renal fibrosis. Discoidin domain receptor 1 (DDR1) is a nonintegrin tyrosine kinase receptor for collagen implicated in cell adhesion, proliferation, and extracellular matrix remodeling. We have previously demonstrated that transgenic mice lacking DDR1 are protected from hypertension-associated renal fibrosis. The purpose of this study was to determine the role of DDR1 in renal inflammation and fibrosis related to primitive tubulointerstitial injury. After 12 days of unilateral ureteral obstruction (UUO), kidney histopathologic and real-time quantitative PCR analyses were performed in DDR1(-/-) and wild-type mice. DDR1 expression was strongly increased in the obstructed kidney. Wild-type mice developed important perivascular and interstitial inflammation and fibrosis. In comparison, DDR1(-/-) mice displayed reduced accumulation of fibrillar collagen and transforming growth factor beta expression. F4/80(+) cell count and proinflammatory cytokines were remarkably blunted in DDR1(-/-) obstructed kidneys. Leukocyte rolling and adhesion evaluated by intravital microscopy were not different between DDR1(-/-) and wild-type mice. Importantly, macrophages isolated from DDR1(-/-) mice presented similar M1/M2 polarization but displayed impaired migration in response to monocyte chemoattractant protein-1. Together, these data suggest that DDR1 plays an important role in the pathogenesis of renal disease via enhanced inflammation. Inhibition of DDR1 expression or activity may represent a novel therapeutic target against the progression of renal diseases. PMID- 21640972 TI - Pathogenesis of Influenza A/H5N1 virus infection in ferrets differs between intranasal and intratracheal routes of inoculation. AB - Most patients infected with highly pathogenic avian influenza A/H5N1 virus develop severe pneumonia resulting in acute respiratory distress syndrome, with extrarespiratory disease as an uncommon complication. Intranasal inoculation of ferrets with influenza A/H5N1 virus causes lesions in both the respiratory tract and extrarespiratory organs (primarily brain). However, the route of spread to extrarespiratory organs and the relative contribution of extrarespiratory disease to pathogenicity are largely unknown. In the present study, we characterized lesions in the respiratory tract and central nervous system (CNS) of ferrets (n = 8) inoculated intranasally with influenza virus A/Indonesia/5/2005 (H5N1). By 7 days after inoculation, only 3 of 8 ferrets had a mild or moderate bronchointerstitial pneumonia. In contrast, all 8 ferrets had moderate or severe CNS lesions, characterized by meningoencephalitis, choroiditis, and ependymitis, and centered on tissues adjoining the cerebrospinal fluid. These findings indicate that influenza A/H5N1 virus spread directly from nasal cavity to brain, and that CNS lesions contributed more than pulmonary lesions to the pathogenicity of influenza A/H5N1 virus infection in ferrets. In comparison, intratracheal inoculation of ferrets with the same virus reproducibly caused severe bronchointerstitial pneumonia. The method of virus inoculation requires careful consideration in the design of ferret experiments as a model for influenza A/H5N1 in humans. PMID- 21640973 TI - Selective stimulation of VEGFR2 accelerates progressive renal disease. AB - Vascular endothelial growth factor A (VEGF-A) can play both beneficial and deleterious roles in renal diseases, where its specific function might be determined by nitric oxide bioavailability. The complexity of VEGF-A in renal disease could in part be accounted for by the distinct roles of its two receptors; VEGFR1 is involved in the inflammatory responses, whereas VEGFR2 predominantly mediates angiogenesis. Because nondiabetic chronic renal disease is associated with capillary loss, we hypothesized that selective stimulation of VEGFR2 could be beneficial in this setting. However, VEGFR2 activation may be deleterious in the presence of nitric oxide deficiency. We systematically overexpressed a mutant form of VEGF-A binding only VEGFR2 (Flk-sel) using an adeno-associated virus-1 vector in wild-type and eNOS knockout mice and then induced renal injury by uninephrectomy. Flk-sel treatment increased angiogenesis and lowered blood pressure in both mouse types. Flk-sel overexpression caused mesangial injury with increased proliferation associated with elevated expression of PDGF, PDGF-beta receptor, and VEGFR2; this effect was greater in eNOS knockout than in wild-type mice. Flk-sel also induced tubulointerstitial injury, with some tubular epithelial cells expressing alpha-smooth muscle actin, indicating a phenotypic evolution toward myofibroblasts. In conclusion, prestimulation of VEGFR2 can potentiate subsequent renal injury in mice, an effect enhanced in the setting of nitric oxide deficiency. PMID- 21640974 TI - Targeting ST2L potentiates CpG-mediated therapeutic effects in a chronic fungal asthma model. AB - IL-33 and its soluble receptor and cell-associated receptor (ST2L) are all increased in clinical and experimental asthma. The present study addressed the hypothesis that ST2L impairs the therapeutic effects of CpG in a fungal model of asthma. C57BL/6 mice were sensitized to Aspergillus fumigatus and challenged via i.t. instillation with live A. fumigatus conidia. Mice were treated with IgG alone, anti-ST2L monoclonal antibody (mAb) alone, CpG alone, IgG plus CpG, or anti-ST2L mAb plus CpG every other day from day 14 to day 28 and investigated on day 28 after conidia. Lung ST2L and toll-like receptor 9 protein expression levels concomitantly increased in a time-dependent manner during fungal asthma. Therapeutic blockade of ST2L with an mAb attenuated key pathological features of this model. At subtherapeutic doses, neither anti-ST2L mAb nor CpG alone affected fungal asthma severity. However, airway hyperresponsiveness, mucus cell metaplasia, peribronchial fibrosis, and fungus retention were markedly reduced in asthmatic mice treated with the combination of both. Whole lung CXCL9 levels were significantly elevated in the combination group but not in the controls. Furthermore, in asthmatic mice treated with the combination therapy, dendritic cells generated significantly greater IL-12p70 with CpG in vitro compared with control dendritic cells. The combination of anti-ST2L mAb with CpG significantly attenuated experimental asthma, suggesting that targeting ST2L might enhance the therapeutic efficacy of CpG during allergic inflammation. PMID- 21640976 TI - Primary bilateral adrenal lymphoma revealed by hemophagocytic syndrome. AB - Primary adrenal lymphoma is rare. It is often bilateral and in most of the cases of B-cell type. The clinical features are various and not specific. We report a case of a 69-year-old woman who had a diffuse large B-cell lymphoma associated with hemophagocytic syndrome. The abdominal imaging reveals the existence of bilateral adrenal hypertrophy. A CT scan-guided biopsy concluded to a diffuse large B-cell lymphoma CD 20-positive associated EBV. The treatment consisted on "CHOP like" chemotherapy associated with rituximab. Primary adrenal lymphoma has a poor prognosis, even more poorly if associated with hemophagocytic syndrome. PMID- 21640975 TI - SHIP-deficient mice develop spontaneous intestinal inflammation and arginase dependent fibrosis. AB - Intestinal fibrosis is a serious complication of Crohn's disease (CD) that can lead to stricture formation, which requires surgery. Mechanisms underlying intestinal fibrosis remain elusive because of a lack of suitable mouse models. Herein, we describe a spontaneous mouse model of intestinal inflammation with fibrosis and the profibrotic role of arginase I. The Src homology 2 domain containing inositol polyphosphate 5'-phosphatase-deficient (SHIP(-/-)) mice developed spontaneous discontinuous intestinal inflammation restricted to the distal ileum starting at the age of 4 weeks. Mice developed several key features resembling CD, including inflammation and fibrosis. Inflammation was characterized by abundant infiltrating Gr-1-positive immune cells, granuloma-like immune cell aggregates that contained multinucleated giant cells, and a mixed type 2 and type 17 helper T-cell cytokine profile. Fibrosis was characterized by a thickened ileal muscle layer, collagen deposition, and increased fibroblasts at the sites of collagen deposition. SHIP(-/-) ilea had increased arginase activity and arginase I expression that was inversely proportional to nitrotyrosine staining. SHIP(-/-) mice were treated with the arginase inhibitor S-(2 boronoethyl)-l-cysteine, and changes in the disease phenotype were measured. Arginase inhibition did not affect the number of immune cell infiltrates in the SHIP(-/-) mouse ilea; rather, it reduced collagen deposition and muscle hyperplasia. These findings suggest that arginase activity is a potential target to limit intestinal fibrosis in patients with CD. PMID- 21640977 TI - Increased thyroidal T4 to T3 conversion in autonomously functioning thyroid adenoma: from euthyroidism to thyrotoxicosis. AB - AIM: The aim was to investigate whether the intrathyroid conversion of T4 to T3 in autonomously functioning thyroid adenoma (AFTA) tissue could influence serum T3 levels and suppression of TSH, especially in patients with borderline thyroid function. PATIENTS AND METHODS: In ten patients with AFTA, thyroidal conversion of T4 to T3 was investigated in nodular and paranodular, TSH-suppressed tissue. All patients had normal serum T4 and suppressed TSH. Serum T3 was normal in six, and borderline or slightly increased in four. AFTA and paranodular tissues were surgically removed and frozen at -70 degrees C, then homogenized in a glass homogenizer, centrifuged at 100,000*g, and particulate fraction collected as a pellet. Analysis mixture consisted of thyroid enzyme suspension in 50 MUmol/L TRIS buffer with 5 MUmol DTT and 200 MUL 1.3 MUmol T4. Incubation was performed at 37 degrees C and the generation of T3 measured after 5, 10, 20 and 40 minutes respectively. RESULTS: T3 production (pmol/mg protein) was significantly higher in AFTA than in paranodular tissues (8.8 1.2/Mean +/- SE/vs. 1.8 +/- 0.2; p<0.01), and excessively high (9.8, 14.1, 14.2 and 15.0) in four patients with borderline or slightly supranormal serum T3. A significant correlation was found between serum T3 concentrations and T3 generation (T4 conversion) in AFTA tissues. CONCLUSION: Results suggest that increased thyroidal T4 to T3 conversion in AFTA tissue could be involved in an increased delivery of T3, increased serum T3 and suppressed serum TSH, particularly in patients with the disease evolving from euthyroid to an early hyperthyroid phase. PMID- 21640978 TI - 45,X/46,XY mosaicisme: report of five cases and clinical review. AB - INTRODUCTION: The mosaicism 45, X/46, XY is a gonosomal abnormality characterized by a broad phenotypic spectrum, ranging from women with or without Turner syndrome stigmata, to men apparently normal, passing by the ambiguous phenotypes with variable virilisation of external genitalia. From the histological point of view, several situations may arise. PATIENTS AND METHODS: We analyzed the clinical, hormonal, sonographic, and genitographics data, as well as peroperative and histological findings for five cases of mosaicism 45, X/46, XY, and we discussed treatment performed. RESULTS: The mean age of patients was 6.6 years, two had a female phenotype with clitoral hypertrophy (one of them had Turner syndrome stigmata), one had a normal male phenotype with bilateral cryptorchidism and two had an ambiguity of external genitalia assigned to male. Short stature was noted for four patients. Surgical exploration concluded to the diagnosis of mixed gonadal dysgenesis for four of our patients. No cases of gonadoblastoma have been reported, for girls a prophylactic gonadectomy was performed, for boys the streak gonad was resected and the dysgenetic testis biopsied and preserved, subject for constant monitoring. CONCLUSION: This heterogeneity indicate the importance of an accurate clinical and histological evaluation of any patient presenting with 45, X/46,XY mosaicism. PMID- 21640979 TI - Odontoblasts in developing, mature and ageing rat teeth have multiple phenotypes that variably express all nine voltage-gated sodium channels. AB - OBJECTIVE: Our goal was to evaluate the expression patterns for voltage gated sodium channels in odontoblasts of developing and mature rat teeth. DESIGN: We analysed immunoreactivity (IR) of the alpha subunit for all nine voltage gated sodium channels (Nav1.1-1.9) in teeth of immature (4 weeks), young adult (7 weeks), fully mature adult (3 months), and old rats (6-12 months). We were interested in developmental changes, crown/root differences, tetrodotoxin sensitivity or resistance, co-localization with nerve regions, occurrence in periodontium, and coincidence with other expression patterns by odontoblasts such as for transient receptor potential A1 (TRPA1). RESULTS: We found that Nav1.1-1.9 IR each had unique odontoblast patterns in mature molars that all differed from developmental stages and from incisors. Nav1.4- and Nav1.7-IR were intense in immature odontoblasts, becoming limited to specific zones in adults. Crown odontoblasts lost Nav1.7-IR and gained Nav1.8-IR where dentine became innervated. Odontoblast staining for Nav1.1- and Nav1.5-IR increased in crown with age but decreased in roots. Nav1.9-IR was especially intense in regularly scattered odontoblasts. Two tetrodotoxin-resistant isoforms (Nav1.5, Nav1.8) had strong expression in odontoblasts near dentinal innervation zones. Nav1.6-IR was concentrated at intercusp and cervical odontoblasts in adults as was TRPA1-IR. Nav1.3-IR gradually became intense in all odontoblasts during development except where dentinal innervation was dense. CONCLUSIONS: All nine voltage-gated sodium channels could be expressed by odontoblasts, depending on intradental location and tooth maturity. Our data reveal much greater complexity and niche-specific specialization for odontoblasts than previously demonstrated, with implications for tooth sensitivity. PMID- 21640981 TI - Prefrontal cortex recruitment during naturalistic remote memory: a factorial block-event fMRI study. AB - Most classical neuroimaging studies of human memory require the subject to follow task instructions, discriminate stimuli, make a decision and, finally, press a button to respond. To which extent does such a cognitive sequence influence neural correlates of recognition memory especially with respect to the prefrontal cortex (PFC) is not clear yet. We set up a naturalistic recognition task. Personal photographs were mixed with photographs from family albums of people who were not known to the participant. The subjects alternated a recognition task (recognition and response) with a mere observational one (simple recognition). This factorial block-event functional MRI design allowed us to disentangle the regions related to memory (personal photographs vs. distracters under both conditions) from those related to the response (responding vs. observing) as well as to examine the interaction between the two factors. Only medial and left orbito-frontal regions were transiently active during successful recognition in the two task conditions. Memory and responding interacted in the left lateral PFC within the middle and inferior gyrus. These regions were more active for personal photographs than for distracters only when the subject had to respond. Sustained activations spread bilaterally whatever the task. Such a design that encompasses personal photographs, an observation period for recognition as well as a mixed design may have led to a better isolation of the neural network underlying remote autobiographical memory. Recruitment within the PFC during retrieval was only limited to its medial and left anterior parts. Sustained activation did not differ between the discrimination and the observation period. This original design might be valuable for further dissociation between decision, self, autobiographical memory and the PFC. PMID- 21640980 TI - Neurotoxic factors released by stimulated human monocytes and THP-1 cells. AB - Activated monocytes/macrophages are known to release toxic materials. Identification of these materials is important for developing more effective treatments for inflammatory disorders where self attack occurs. We stimulated human monocytes and THP-1 cells with LPS/IFNgamma and measured the toxic effects of their conditioned media against differentiated human NT-2 cells. Their cytotoxicity, as measured by LDH release, was reduced by half when their conditioned media was passed through a 3kDa cutoff filter, indicating an equal division between high and low molecular weight materials. When the high molecular weight components tumor necrosis factor-alpha (TNFalpha), interleukin-1beta (IL 1beta), and IL-6 were removed from the conditioned medium by specific antibodies, the toxicity was reduced by 37-38%. When prostaglandin production was blocked by treatment with the COX inhibitors acetylsalicylic acid and ibuprofen, toxicity was reduced by 15-16%. When oxygen free radical production was blocked by the NADPH inhibitor diphenylene iodonium (DPI) the toxicity was reduced by 17-18%. Treatment with the nitric oxide scavenger carboxy-phenyl tetramethylimidazolineoxyl-oxide, or the NOS inhibitor N(G)-monomethylene-l arginine, attenuated the toxicity by about 20%. Removal of released glutamate by glutamate decarboxylase also attenuated the toxicity by 12-13%. In combination, these treatments reduced the toxicity by approximately 50% accounting for the low molecular weight component toxicity. About 10% of the overall toxicity, which was associated with the high molecular weight component, was not identified. Optimal antiinflammatory therapy may require combined suppression of these identified toxin-generating pathways as well as relatively minor pathways yet to be identified. PMID- 21640982 TI - A comparative study of curcuminoids to measure their effect on inflammatory and apoptotic gene expression in an Abeta plus ibotenic acid-infused rat model of Alzheimer's disease. AB - Alzheimer's disease (AD) is a neurodegenerative disorder, which depicts features of chronic inflammatory conditions resulting in cellular death and has limited therapeutic options. We aimed to explore the effect of a curcuminoid mixture and its individual components on inflammatory and apoptotic genes expression in AD using an Abeta+ibotenic acid-infused rat model. After 5 days of treatment with demethoxycurcumin, hippocampal IL-1beta levels were decreased to 118.54 +/- 47.48 and 136.67 +/- 31.96% respectively at 30 and 10mg/kg, compared with the amyloid treated group (373.99 +/- 15.28%). After 5 days of treatment, the curcuminoid mixture and demethoxycurcumin effectively decreased GFAP levels in the hippocampus. When studied for their effect on apoptotic genes expression, the curcuminoid mixture and bisdemethoxycurcumin effectively decreased caspase-3 level in the hippocampus after 20 days of treatment, where bisdemethoxycurcumin showed a maximal rescuing effect (92.35 +/- 3.07%) at 3mg/kg. The curcuminoid mixture at 30 mg/kg decreased hippocampal FasL level to 70.56 +/- 3.36% after 5 days of treatment and 19.01 +/- 2.03% after 20 days. In the case of Fas receptor levels, demethoxycurcumin decreased levels after 5 days of treatment with all three doses showing a maximal effect (189.76 +/- 15.01%) at 10mg/kg. Each compound was effective after 20 days in reducing Fas receptor levels in the hippocampus. This study revealed the important effect of curcuminoids on genes expression, showing that, each component of the curcuminoid mixture distinctly affects gene expression, thus highlighting the therapeutic potential of curcuminoids in AD. PMID- 21640984 TI - Synergistic influence of sucrose and abscisic acid on the genes involved in starch synthesis in maize endosperm. AB - Starch is the major carbon reserve in plant storage organs, the synthesis of which is orchestrated by four major enzymes, ADP-glucose pyrophosphorylase, starch synthase, starch-branching enzyme and starch-debranching enzyme. There is much information available on the function of these key enzymes; however, little is known about their transcriptional regulation. In order to understand the transcriptional regulation of starch biosynthesis, the expression profiles of 24 starch genes were investigated in this work. The results showed major transcriptional changes for 15 of the 24 starch genes observed in maize endosperm, most of which are elevated at the early and middle stages of the developing endosperm. Sucrose, abscisic acid (ABA) and indole-3-acetic acid (IAA) had a significant correlation with the expression of 15 genes, indicating that sugars and phytohormones might take part in the regulation of starch synthesis. Also, we found that there is interaction of abscisic acid and sucrose on the regulation of the expression of these genes. PMID- 21640983 TI - Cortical and subcortical changes in typically developing preadolescent children. AB - There is evidence that abnormal cerebral development during childhood is a risk factor for various cognitive and psychiatric disorders. There is not, however, sufficient normative data available on large samples of typically developing children, especially within the narrow preadolescent age range. We analyzed high resolution MRI images from 126 normally developing children between ages 6 and 10 years. Age related differences in cortical thickness and in the volumes of major subcortical structures were assessed. Thinner cortices were observed in the occipital, parietal and somatosensory regions as well as in distinct regions of the temporal and frontal lobes with increasing age. Among the major subcortical structures analyzed in this study, only the thalamus showed increased volume with age after accounting for intracranial volume. Within the age range studied age related cortical and subcortical differences were similar for boys and girls except for the right insula, where girls showed a slight increase in thickness with age. The findings reveal age-associated changes in brain anatomy, providing information about the trajectory of normal brain development during late childhood. PMID- 21640985 TI - Do CD4+ Foxp3+ Treg cells correlate with transplant outcomes: a systematic review on recipients of solid organ transplantation. AB - Regulatory T cells (Tregs) are considered to be critical for the induction of transplant tolerance. Tregs counts were measured in blood, biopsy and urine sample after transplantation in many studies. Although not unanimous, some studies have suggested that Tregs is associated with better outcome and can also serve as an immune marker to predict the individual risk of rejection and identify tolerant patients. In this study, we systematically reviewed the correlation between Tregs and transplant outcomes, identifying if Tregs can predict transplant rejection and tolerance. A total of 22 articles were included and assessed, the results showed that Tregs in recipients are helpful to maintain a stable graft function, reduce acute/chronic rejection rate. And the Tregs in graft and urine, rather than in PBL, may have a better diagnostic value for transplant outcomes. However, since the low quality of included studies, results may be influenced by bias. More high quality studies with bigger sample size are still needed in future. PMID- 21640986 TI - [Complications of distal intestinal occlusion treatment with endoluminal implants]. AB - INTRODUCTION: The high morbidity and mortality of emergency surgery, has led to the use of endoluminal self-expanding metal implants (stents) in the management of intestinal occlusion. The purpose of this study was to review the results of the management of intestinal occlusion treatment in a Colorectal Surgery Unit in those patients who had a stent implant, and the relationship between chemotherapy and complications. MATERIAL AND METHODS: A retrospective study was carried out on patients treated with a stent in a university hospital between 2004 and 2010. RESULTS: A total of 93 patients were treated, of which 77 were considered palliative for a stage IV neoplasm of the colon with non-resectable metastases or due to a performance status > 2. Other indications were 7 ASA IV patients with acute renal failure, 6 with benign disease, and 3 due to other causes. The technical and clinical success of the procedure was 93.5% and 78.5%, respectively. Delayed occlusion was 19.3% and perforation 6.4%. There was migration (2.1%) and intestinal bleeding (2.1%) and 1.1% with tenesmus. No significant differences were seen between complications and chemotherapy. The overall mortality was 17.2%. CONCLUSIONS: Stents, as a definitive treatment option in palliative patients with and without chemotherapy, is an alternative treatment that is not exempt from complications. We believe that in patients with mortality risk factors and patients with tumours with non-resectable metastases it could be the initial treatment of choice. PMID- 21640987 TI - [Distal ileal perforation in a 30-week pregnant woman with a history of total colectomy due to familial adenomatous polyposis]. PMID- 21640988 TI - [Mucinous adenocarcinoma over an anorectal fistula in a patient with Crohn's diseases]. PMID- 21640989 TI - [Uterine and sigmoid colon perforation due to an intrauterine device]. PMID- 21640990 TI - Multichannel least-squares linear regression provides a fast, accurate, unbiased and robust estimation of Granger causality for neurophysiological data. AB - The most common method for calculating Granger causality requires the fitting of a system of autoregressive equations to multiple interrelated signals. Historically, the Levinson, Wiggins, Robinson (LWR) algorithm and the least squares linear regression (LSLR) approach are the most widely used methods for fitting these autoregressive equations. In this manuscript we compare these algorithms head-to-head. LSLR, as implemented using the Dynamic Autoregressive Neuromagnetic Causal Imaging (DANCI) method, was faster, and produced better residual error, normality, independence, and autocorrelation functions when analyzing real magnetoencephalography signals. Simulations demonstrated that the accuracy of LSLR was much higher than the LWR method and that the LSLR method, at least as implemented by DANCI, could accurately resolve the causal connectivity of 50 interrelated signals. We conclude that the multichannel LSLR method, as implemented by DANCI, can accurately calculate the interdependencies among multiple signals and has the potential to accurately calculate Granger causality for large-scale neurophysiological networks. PMID- 21640991 TI - Incidence and diagnosis of heparin-induced thrombocytopenia (HIT) in patients with traumatic injuries treated with unfractioned or low-molecular-weight heparin: a literature review. AB - INTRODUCTION: The incidence of thromboembolic complications after major traumatic injuries is high (>50%). Thromboprophylaxis, often by low-molecular-weight heparin (LMWH) or unfractioned heparin (UH) is therefore routinely administered. Thromboprophylaxis is also advised after immobilisation for isolated lower leg injuries. Heparin induced thrombocytopenia (HIT) is a rare but very serious immune mediated complication of treatment with LMWH, which can cause potentially fatal thromboembolism. In the general medical and surgical population the incidence of HIT is 0.2%. Little is known about the incidence of HIT and value of screening in trauma patients and in isolated lower extremity injuries. Therefore, we performed a systematic literature review. METHODS: The online databases Medline and EMBASE were searched independently by two authors. Manuscripts were selected for analysis by quantitative and qualitative selection. After eliminating duplicate articles and irrelevant studies, seven relevant papers reporting on the incidence of HIT in trauma patients were identified and two studies reported the incidence of HIT in patients with lower leg injuries. RESULTS: The selected papers varied in study design: three randomised controlled trials, three cohort studies and one case report were identified. The methodological quality of the studies varied. In a total population of 1920 patients, HIT was identified in seven patients (0.36%). Pooling of data was impossible due to heterogeneity in study design and populations. No HIT was reported in 826 patients with lower extremity injuries, requiring immobilisation. DISCUSSION: Only a few studies have reported on the incidence of HIT in trauma patients who receive prophylactic LMWH. In the heterogenous populations of the available studies, the incidence of HIT appears to be very low and comparable to other patient populations. There is hardly any literature on the incidence of HIT in patients with isolated lower leg injuries receiving LMWH, but incidence seems to be very low. CONCLUSION: The incidence of HIT in trauma patients who receive LMWH thromboprophylaxis appears to be low (0.36%). Incidence of HIT in patients with isolated lower leg injuries receiving LMWH seems very low. Monitoring of platelet count could be considered in hospitalised patients with a high risk for development of HIT. A pre-test scoring system may identify these patients. PMID- 21640992 TI - Living through the death of a child: a qualitative study of bereaved parents' experiences. AB - DESIGN: Cross-sectional qualitative study. DATA SOURCES: Interviews with purposeful sample of 25 recently bereaved parents. METHODS: Semi-structured in depth interviews. RESULTS: Four analytically distinct processes were identified in the responses of parents to the death of a child. These are referred to as 'piloting', 'providing', 'protecting' and 'preserving'. Regardless of individual circumstances, these processes were integral to all parents' coping, enabling an active 'doing' for their child and family throughout the trajectory of their child's illness and into bereavement. CONCLUSIONS: Facilitating the capacity of parents to 'do' is central to coping with the stress and uncertainty of living through the death of a child. The provision of informational, instrumental and emotional support by health care professionals in the context of 'doing' is core to quality palliative care. PMID- 21640993 TI - Common variation in the ADAM8 gene affects serum sADAM8 concentrations and the risk of myocardial infarction in two independent cohorts. AB - OBJECTIVE: The single nucleotide polymorphism (SNP) rs2995300 in the metalloproteinase-disintegrin gene ADAM8 has been shown to affect the areas of complicated coronary plaques and the risk of fatal myocardial infarction (MI) in men. This study was set up to further investigate the role of ADAM8 in MI. AIM: To investigate the possible association of the ADAM8 SNPs rs2995300 and rs2275725 with ADAM8 mRNA levels, serum soluble ADAM8 (sADAM8) concentrations, and MI risk. METHODS: Samples from the Finnish cardiovascular study (FINCAVAS, N=2156) and the angiography and genes study (ANGES, N=1000) were genotyped. Serum sADAM8 concentrations were determined with ELISA (N=443). ADAM8 mRNA levels in atherosclerotic plaques were analysed from the tampere vascular study (TVS, N=53) samples. RESULTS: A significantly increased MI risk for carriers of the rs2995300C allele and the rs2275725 A allele was revealed in the meta-analysis of the ANGES and FINCAVAS patient data (OR=1.42, P<0.001 and OR=1.43, P<0.001). The risk increase was comparable to that caused by smoking in these cohorts. The risk allele carriers also had higher sADAM8 serum concentrations. CONCLUSIONS: The risk alleles of the investigated ADAM8 SNPs were associated with elevated sADAM8 serum levels and MI risk. The present results implicate ADAM8 in the development of CVDs and suggest its prognostic and therapeutic potential. PMID- 21640994 TI - Hs-CRP may be associated with white blood cell count in metabolic syndrome patients treated with Ginkgo biloba. PMID- 21640995 TI - Short range stiffness elastic limit depends on joint velocity. AB - Muscles behave as elastic springs during the initial strain phase, indicated as short range stiffness (SRS). Beyond a certain amount of strain the muscle demonstrates a more viscous behavior. The strain at which the muscle transits from elastic- to viscous-like behavior is called the elastic limit and is believed to be the result of breakage of cross-bridges between the contractile filaments. The aim of this study was to test whether the elastic limit, measured in vivo at the wrist joint, depended on the speed of lengthening. Brief extension rotations were imposed to the wrist joint (n=8) at four different speeds and at three different levels of voluntary torque using a servo controlled electrical motor. Using a recently published identification scheme, we quantified the elastic limit from measured joint angle and torque. The results showed that the elastic limit significantly increased with speed in a linear way, indicating to a constant time of approximately 30 ms before cross-bridges break. The implications for movement control of the joint are discussed. PMID- 21640996 TI - Effect of muscle contraction levels on the force-length relationship of the human Achilles tendon during lengthening of the triceps surae muscle-tendon unit. AB - Findings from animal experiments are sometimes contradictory to the idea that the tendon structure is a simple elastic spring in series with muscle fibers, and suggest influence of muscle contraction on the tendon mechanical properties. The purpose of the present study was to investigate the influence of muscle contraction levels on the force-length relationship of the human Achilles tendon during lengthening of the triceps surae muscle-tendon unit. For seven subjects, ankle dorsiflexion was performed without (passive condition) and with contraction of plantar flexor muscles (eccentric conditions, at 3 contraction levels) on an isokinetic dynamometer. Deformation of the Achilles tendon during each trial was measured using ultrasonography. The Achilles tendon force corresponding to the tendon elongation of 10mm in the passive condition was significantly smaller than those in the eccentric conditions (p<0.05 or p<0.01). Within the eccentric conditions, the Achilles tendon force corresponding to the tendon elongation of 10mm was significantly greater in the maximal contraction level than those in submaximal eccentric conditions (p<0.05 or p<0.01). In addition, the tendon stiffness was greater in higher contraction levels (p<0.05 or p<0.01). Present results suggest that the human tendon structure is not a simple elastic spring in series with muscle fibers. PMID- 21640997 TI - Savinase proteolysis of insulin Langmuir monolayers studied by surface pressure and surface potential measurements accompanied by atomic force microscopy (AFM) imaging. AB - The mechanism of the enzymatic action of Savinase on an insulin substrate organized in a monolayer at the air-water interface was studied. We followed two steps experimental approach classical surface pressure and surface potential measurements in combination with atomic force microscopy imaging. Utilizing the barostat surface balance, the hydrolysis kinetic was followed by measuring simultaneously the decrease in the surface area and the change of the surface potential versus time. The decrease in the surface area is a result of the random scission of the peptide bonds of polypeptide chain, progressively appearance of amino acid residues, and their solubilization in the aqueous subphase. The interpretation of the surface potential data was based on the contribution of the dipole moments of the intact and broken peptide groups which remain at the interface during the proteolysis. An appropriate kinetic model for the Savinase action was applied, and the global kinetic constant was obtained. The application of the AFM revealed the state of the insulin monolayers before and after the Savinase action. The comparison of the topography of the films and the roughness analysis showed that insulin Langmuir-Blodgett (LB) films transferred before the enzyme action were flat, while at the end of hydrolysis, roughness of films has increased and the appearance of 3D structures was observed. PMID- 21640998 TI - Sensitive protein microarray synergistically amplified by polymer brush-enhanced immobilizations of both probe and reporter. AB - Great challenge remains to continuously improve sensitivity of protein microarrays for broad applications. A copolymer brush is in situ synthesized on both substrate and silica nanoparticle (SNP) surface to efficiently immobilize probe and reporter protein respectively for synergistic amplification of protein microarray signals. As a demonstration, sandwich immunoassay for a cancer biomarker carcinoembryonic antigen (CEA) detection is performed on microarray platform, showing a limit of detection (LOD) of 10 pg/ml and dynamic range of 10 pg/ml to 100 ng/ml. Two orders improvement of LOD is achieved in comparison to the small crosslinker-activated substrate. The improved sensitivity is attributed to not only the high immobilization amount of both probe and reporter but also the favorite protein binding orientations offered by the flexible brushes. This work provides a universal approach to inexpensively and significantly improve protein microarray sensitivity. PMID- 21640999 TI - Template-assisted encapsulation of fluorinated silanes in silica films for sustained hydrophobic-oleophobic functionality. AB - This work explores the use of templated silica films as scaffolds for encapsulating surface-segregating functional organic moieties as a mesoscopically dispersed phase with the goal of imparting sustained functionality. Block copolymer surfactant templated hydrophobic-oleophobic fluorinated silica films were synthesized via sol-gel co-condensation and coated on glass substrates. Fluorosilane and surfactant template concentrations were varied, and coating surface properties measured before and after abrasion of the top surface. Surface physical and chemical properties were investigated using XPS and contact angle measurements. Nitrogen adsorption porosimetry and TEM were used to examine the effect of templating and fluorosilane encapsulation on the surrounding silica framework. The results show that surfactant template concentration may be used to tune the dispersion of the fluorosilane-rich phase within the silica film in order to allow exposed surfaces to maintain much of the original functionality of the pristine top surface. PMID- 21641000 TI - Copying skills in relation to word reading and writing in Chinese children with and without dyslexia. AB - Because Chinese character learning typically relies heavily on rote character copying, we tested independent copying skill in third- and fourth-grade Chinese children with and without dyslexia. In total, 21 Chinese third and fourth graders with dyslexia and 33 without dyslexia (matched on age, nonverbal IQ, and mother's education level) were given tasks of copying unfamiliar print in Vietnamese, Korean, and Hebrew as well as tests of word reading and writing, morphological awareness, rapid automatized naming (RAN), and orthographic processing. All three copying tasks distinguished dyslexic children from nondyslexic children with moderate effect sizes (.67-.80). Zero-order correlations of the three copying tasks with dictation and reading ranged from .37 to .58. With age, Raven's, group status, RAN, morphological awareness, and orthographic measures statistically controlled, the copying tasks uniquely explained 6% and 3% variance in word reading and dictation, respectively. Results suggest that copying skill itself may be useful in understanding the development and impairment of literacy skills in Chinese. PMID- 21641001 TI - Do young and old preschoolers exhibit response bias due to different mechanisms? Investigating children's response time. AB - Previous studies have suggested that younger preschoolers exhibit a yes bias due to underdeveloped cognitive abilities, whereas older preschoolers exhibit a response bias due to other factors. To test this hypothesis, we investigated the response latency to yes-no questions pertaining to familiar and unfamiliar objects in 3- to 6-year-olds. The 3-year-olds exhibited a strong yes bias for both objects, and their response latency was significantly shorter than that of the 6-year-olds. The 4- and 5-year-olds did not exhibit any response bias, whereas the 6-year-olds exhibited a nay-saying bias (a response bias to say "no") for unfamiliar objects. Overall, children's response bias scores were stronger for familiar objects than for unfamiliar objects. The results suggest that 3-year olds exhibit a yes bias automatically, but 5- and 6-year-olds are sensitive to question context. PMID- 21641002 TI - Effects of serum Brain Derived Neurotrophic Factor on exercise augmentation treatment of depression. AB - BACKGROUND: Brain Derived Neurotrophic Factor (BDNF) has potential as a biomarker of depression treatment because serum BDNF in depressed human subjects is decreased and normalizes with treatment. The relationship between serum BDNF and exercise treatment of depression is not known. The Treatment with Exercise Augmentation for Depression (TREAD) study examined dosed exercise augmentation treatment of partial responders to antidepressants. Serum BDNF in TREAD subjects was analyzed to understand its relationship with exercise training. METHODS: Subjects were randomized to high (16 kcal/kg/week or KKW) or low (4 KKW) energy expenditure exercise over 12 weeks. Actual kcal/week expended and IDS-C scores were collected weekly. One hundred four subjects in TREAD provided baseline blood samples; a subset of 70 subjects also provided week 12 samples. Serum BDNF was determined using ELISA. Correlations were examined between change in BDNF and 1) mean kcal/week expended, and 2) change in IDS-C score. Mixed-effects ANOVA examined the effect of baseline BDNF on outcome. RESULTS: Resting serum BDNF was stable and did not correlate with energy expenditure (p = 0.15) or IDS-C improvement (p = 0.89). Subjects entering the study with higher BDNF improved more rapidly on the IDS-C (p = 0.003). LIMITATIONS: Serum may not be the most sensitive blood fraction in which to measure BDNF change. Pre-treatment with medication may mask exercise effect on BDNF. CONCLUSIONS: These results suggest that change in serum BDNF does not reflect efficacy of exercise augmentation treatment of MDD. Instead BDNF may function as an augmentation moderator. Pre treatments that raise BDNF may improve the efficacy of exercise treatment of MDD. PMID- 21641003 TI - High frequency stimulation of the subthalamic nucleus increases c-fos immunoreactivity in the dorsal raphe nucleus and afferent brain regions. AB - High frequency stimulation (HFS) of the subthalamic nucleus (STN) is the neurosurgical therapy of choice for the management of motor deficits in patients with advanced Parkinson's disease, but this treatment can elicit disabling mood changes. Our recent experiments show that in rats, HFS of the STN both inhibits the firing of 5-HT (5-hydroxytryptamine; serotonin) neurons in the dorsal raphe nucleus (DRN) and elicits 5-HT-dependent behavioral effects. The neural circuitry underpinning these effects is unknown. Here we investigated in the dopamine denervated rat the effect of bilateral HFS of the STN on markers of neuronal activity in the DRN as well as DRN input regions. Controls were sham-stimulated rats. HFS of the STN elicited changes in two 5-HT-sensitive behavioral tests. Specifically, HFS increased immobility in the forced swim test and increased interaction in a social interaction task. HFS of the STN at the same stimulation parameters, increased c-fos immunoreactivity in the DRN, and decreased cytochrome C oxidase activity in this region. The increase in c-fos immunoreactivity occurred in DRN neurons immunopositive for the GABA marker parvalbumin. HFS of the STN also increased the number of c-fos immunoreactive cells in the lateral habenula nucleus, medial prefrontal cortex but not significantly in the substantia nigra. Collectively, these findings support a role for circuitry involving DRN GABA neurons, as well as DRN afferents from the lateral habenula nucleus and medial prefrontal cortex, in the mood effects of HFS of the STN. PMID- 21641004 TI - Behavioral changes in early ALS correlate with voxel-based morphometry and diffusion tensor imaging. AB - BACKGROUND: Amyotrophic lateral sclerosis (ALS) is a multisystem disorder with impairment of frontotemporal functions such as cognition and behavior, but the behavioral changes associated with ALS are not well defined. METHODS: Twenty-one consecutive patients with sporadic ALS and 21 control subjects participated in the study. The Frontal System Behavior Scale (FrSBe) was used to assess behavioral change. Voxel-based morphometry (VBM) and voxel-based analysis of diffusion tensor images (DTI) were performed to explore the associations of brain degeneration with behavior. All patients were evaluated before the notification of ALS. RESULTS: FrSBe scores of ALS patients before notification were significantly increased compared to those of control subjects. Moreover, the FrSBe Apathy score of ALS patients significantly changed from pre- to post illness (P<0.001). The severity of apathy was significantly correlated with atrophy in the prefrontal cortex, especially in the orbitofrontal (P=0.006) and dorsolateral prefrontal (P=0.006) cortices in VBM, and in the right frontal gyrus (P<0.001) in DTI. CONCLUSIONS: ALS patients exhibited apathy during the early course of the illness, the severity of which was significantly associated with frontal lobe involvement. These findings support the view that a continuum exits between ALS and frontotemporal dementia. PMID- 21641005 TI - A bovine hemoglobin-based oxygen carrier as pump prime for cardiopulmonary bypass: reduced systemic lactic acidosis and improved cerebral oxygen metabolism during low flow in a porcine model. AB - OBJECTIVES: Cerebral ischemia can occur during cardiopulmonary bypass, especially during low flow. HBOC-201 (OPK Biotech, Cambridge, Mass) is a hemoglobin-based oxygen-carrying solution that enhances oxygen delivery. This project evaluated the benefits on total body and cerebral oxygen delivery and consumption using HBOC-201 during cardiopulmonary bypass. METHODS: Twelve immature swine were assigned to one of 2 groups. One group used HBOC-201 in pump prime, and the other used donor porcine blood. Cardiopulmonary bypass was initiated and then flow was serially decreased from 100% to 75%, to 50%, and then back to full flow. At each interval, (15)O positron emission tomographic analysis was performed, and blood was collected. Total body and cerebral oxygen delivery and consumption were calculated. Statistical analysis was performed with a Tukey-Kramer adjusted P value based on a repeated measures linear model on log-transformed data. RESULTS: Total and plasma hemoglobin levels were higher in the HBOC-201 group. Oxygen delivery and consumption were not statistically different but did tend to be higher in the HBOC-201 group. Mixed venous saturation was lower in the HBOC-201 group but not significant. Mild metabolic acidosis with increased lactate levels developed in the blood group. Mean cerebral blood flow decreased in both groups when total flow was 50%. In the HBOC-201 group cerebral oxygen metabolism was maintained. CONCLUSIONS: The addition of HBOC-201 for cardiopulmonary bypass appears to improve oxygen use and minimize anaerobic metabolism. Cerebral oxygen use was preserved in the HBOC-201 group, even during decrease in blood flow. These findings support the reported improved oxygen-unloading properties of HBOC 201 and might provide a benefit during cardiopulmonary bypass. PMID- 21641006 TI - [Our experience in the diagnosis and treatment of cerebral pseudoaneurysms]. AB - OBJECTIVE: To present our experience in the diagnosis and intravascular treatment of cerebral pseudoaneurysms. MATERIAL AND METHODS: We present 11 pseudoaneurysms (2 traumatic, 2 mycotic, 3 iatrogenic, and 4 with other causes). We analyze the methods and diagnostic criteria, radiological and clinical outcome, the criteria used in making decisions about treatment, the method of treatment, and the complications. RESULTS: Digital subtraction angiography is the gold standard for the diagnosis of cerebral pseudoaneurysms; the diagnostic criteria in the literature include: aneurysms with early morphological changes and distal aneurysms or proximal aneurysms associated with another distal one, in the context of the right symptoms and signs. In the nine patients treated with endovascular techniques, the treatment objective was achieved and rebleeding did not occur. CONCLUSIONS: In cases with clinical suspicion of a pseudoaneurysm, the patient should undergo angiography. This is especially important in patients with inexplicable cerebral hemorrhage and in those with septicemia. CT angiography and MR angiography have good diagnostic accuracy and can replace conventional angiography. However, the treatment of choice is endovascular and treatment should not be delayed unless access to the pseudoaneurysm is impeded, usually due to severe cerebral vasospasm. PMID- 21641007 TI - [Diastolic retrograde arterial flow: preliminary report]. AB - OBJECTIVE: Holodiastolic arterial blood flow is associated with pathological conditions. Nevertheless, we have observed that lifting the arm at an angle greater than the horizontal causes holodiastolic arterial blood flow in the brachial artery in normal patients. Thus, we decided to assess the frequency and characteristics of this phenomenon. MATERIAL AND METHODS: Ten volunteers (7 women) aged 43 +/- 17 years participated in the study. We used an ultrasound scanner with a 12 MHz probe to analyze the brachial artery. The examination included: a) Baseline measurements in the supine position; b) measurements during three minutes with the arm raised, and c) a measurement sixty seconds after lowering the arm to the supine position in which the baseline measurements had been obtained. RESULTS: We observed mid- and end-diastolic retrograde flow in 8/10 patients when their arms were raised. No mid- or end-diastolic retrograde flow was observed in the baseline measurements or after the arm was lowered to the supine position (p=0.0007). The minimum diastolic velocity was significantly higher in the measurements obtained with the arm raised than in the supine position before or after arm raising (-13.5 +/- 4.9 cm/s vs. -2.38 +/- 7.5 cm/s, p<0.05 and -13.5 +/- 4.9 cm/s vs. -4.6 +/- 5.2 cm/s, p<0.05, respectively). The modified resistance index was significantly higher when the arm was raised (1.20 +/- 0.07 vs. 1.04 +/- 0.15; p<0.05); moreover, the modified resistance index was significantly lower in the measurements obtained after the arm was lowered than in the baseline measurements (1.20 +/- 0.07 vs 1.07 +/- 0.08; p<0.05). CONCLUSION: We conclude that holodiastolic reflux occurs in healthy patients. This physiological phenomenon merits further investigation and can help elucidate previous observations in different pathological conditions. PMID- 21641008 TI - [Training tools and methods in radiodiagnosis]. AB - Training the best radiologist is the tutor's aim. Considering both residents features -whose intention is becoming a radiologist- and the trainer profile, the method simultaneously combines the most suitable tools, in order to reach the aim. The method knowledge and the available tools facilitate the tutor planning and his cooperation in the whole resident learning process. PMID- 21641009 TI - Cortisol, adrenocorticotropic hormone, serotonin, adrenaline and noradrenaline serum concentrations in relation to disease and stress in the horse. AB - No detailed comparative data are available on the hormonal parameters of horses suffering from a number of diseases. The aim of our study was to measure concentrations of cortisol, adrenocorticotropic hormone (ACTH), serotonin, adrenaline and noradrenaline in horses with various diseases and following surgery, to assess the response of the HPA axis and adrenal medulla. Blood samples were obtained from six groups of horses comprising a total of 119 animals as follows: laminitis, acute abdominal syndrome (AAS), castration surgery, acute diseases, chronic diseases and healthy controls. Serum hormonal concentrations were determined for each group for comparison. Statistically significant differences between all groups and controls were found for cortisol, ACTH (except for castration), serotonin and adrenaline concentrations but only in horses with laminitis and AAS for noradrenaline. No statistically significant differences were found between males and females. The largest changes in the pituitary adrenal axis activity occurred mainly in acute diseases, laminitis and in the AAS group. PMID- 21641010 TI - Effects of in ovo vaccination and anticoccidials on the distribution of Eimeria spp. in poultry litter and serum antibody titers against coccidia in broiler chickens raised on the used litters. AB - The present study reports the effects of various field anticoccidial programs on the distribution of Eimeria spp. in poultry litter and serum antibody titers against coccidia in broiler chickens raised on the used litters. The programs included in ovo vaccination and various medications with either chemicals, ionophores, or both. In general, serum samples from these chickens showed anticoccidial antibody titers when tested at days 7 and 14 post hatch with the peak response at day 43. Serum anticoccidial titers were highest in birds fed a non-medicated diet compared with those vaccinated or fed medicated diets. Total number of Eimeria oocysts and the composition of Eimeria spp. present in the litter samples from different treatment groups varied depending on the type of anticoccidial program. Oocyst counts in general ranged from 3.7*10(3) to 7.0*10(4) per g of litter. Importantly, both morphological and molecular typing studies revealed four major predominant Eimeria spp., E. acervulina, E. maxima, E. praecox, and E. tenella in the litter samples. Collectively, these results indicate that the field anticoccidial programs influenced the type and abundance of Eimeria spp. present in the litter samples and also modulated host immune response to Eimeria. PMID- 21641011 TI - Process optimization by decoupled control of key microbial populations: distribution of activity and abundance of polyphosphate-accumulating organisms and nitrifying populations in a full-scale IFAS-EBPR plant. AB - This study investigated the abundance and distribution of key functional microbial populations and their activities in a full-scale integrated fixed film activated sludge-enhanced biological phosphorus removal (IFAS-EBPR) process. Polyphosphate accumulating organisms (PAOs) including Accumulibacter and EBPR activities were predominately associated with the mixed liquor (>90%) whereas nitrifying populations and nitrification activity resided mostly (>70%) on the carrier media. Ammonia oxidizer bacteria (AOB) were members of the Nitrosomonas europaea/eutropha/halophila and the Nitrosomonas oligotropha lineages, while nitrite oxidizer bacteria (NOB) belonged to the Nitrospira genus. Addition of the carrier media in the hybrid activated sludge system increased the nitrification capacity and stability; this effect was much greater in the first IFAS stage than in the second one where the residual ammonia concentration becomes limiting. Our results show that IFAS-EBPR systems enable decoupling of solid residence time (SRT) control for nitrifiers and PAOs that require or prefer conflicting SRT values (e.g. >15 days required for nitrifiers and <5 days preferred for PAOs). Allowing the slow-growing nitrifiers to attach to the carrier media and the faster-growing phosphorus (P)-removing organisms (and other heterotrophs, e.g. denitrifiers) to be in the suspended mixed liquor (ML), the EBPR-IFAS system facilitates separate SRT controls and overall optimization for both N and P removal processes. PMID- 21641012 TI - Effects of humic acid on physical and hydrodynamic properties of kaolin flocs by particle image velocimetry. AB - The physical and hydrodynamic properties of kaolin flocs including floc size, strength, regrowth, fractal structure and settling velocity were investigated by in situ particle image velocimetry technique at different humic acid concentration. Jar-test experimental results showed that the adsorbed humic acid had a significant influence on the coagulation process for alum and ferric chloride. Kaolin flocs formed with the ferric chloride were larger and stronger than those for alum at same humic acid concentration. Floc strength and regrowth were estimated by strength factor and recovery factor at different humic acid concentration. It was found that the increased humic acid concentration had a slight influence on the strength of kaolin flocs and resulted in much worse floc regrowth. In addition, the floc regrowth after breakage depended on the shear history and coagulants under investigation. The changes in fractal structure recorded continuously by in situ particle image velocimetry technique during the growth-breakage-regrowth processes provided a supporting information that the kaolin flocs exhibited a multilevel structure. It was proved that the increased humic acid concentration resulted in decrease in mass fractal dimension of kaolin flocs and consequently worse sedimentation performance through free-settling and microbalance techniques. PMID- 21641013 TI - Activation of the phosphatidylinositol 3'-kinase/AKT pathway in neuroblastoma and its regulation by thioredoxin 1. AB - Neuroblastoma is a malignant pediatric tumor with poor survival. The phosphatidylinositol 3'-kinase/AKT pathway is a crucial regulator of cellular processes including apoptosis. Thioredoxin 1, an inhibitor of tumor-suppressor phosphatase and tensin homolog, is overexpressed in many tumors. The objective of this study was to explore phosphatidylinositol 3'-kinase/AKT pathway activation and regulation by thioredoxin 1 to identify potential therapeutic targets. Immunohistochemical analysis was done on tissue microarrays from tumor samples of 101 patients, using antibodies against phosphatidylinositol 3'-kinase, AKT, activated AKT, phosphatase and tensin homolog, phosphorylated phosphatase and tensin homolog, thioredoxin 1, epidermal growth factor receptor, vascular endothelial growth factor and receptors (vascular endothelial growth factor 1 and vascular endothelial growth receptor 2), platelet-derived growth factor receptors, insulin-like growth factor 1 receptor, neurotrophic tyrosine kinase receptor type 2, phosphorylated 70-kd S6 protein kinase, 4E-binding protein 1, and phosphorylated mammalian target of rapamycin. Using 3 neuroblastoma cell lines, we investigated cell viability with AKT-specific inhibitors (LY294002, RAD001) and thioredoxin 1 alone or in combination. We found activated AKT and AKT expressed in 97% and 98%, respectively, of neuroblastomas, despite a high expression of phosphatase and tensin homolog correlated with thioredoxin 1. AKT expression was greater in metastatic than primary tumors. Insulin-like growth factor 1 receptor, tyrosine kinase receptor type 2, vascular endothelial growth receptor 1, and downstream phosphorylated 70-kd S6 protein kinase were correlated with activated AKT. LY294002 and RAD001 significantly reduced AKT activity and cell viability and induced a G(1) cell cycle arrest. Thioredoxin 1 decreased cytotoxicity of AKT inhibitors and doxorubicin, up-regulated AKT activation, and induced cell growth. Thus, vascular endothelial growth receptor 1, tyrosine kinase receptor type 2, insulin-like growth factor 1 receptor, and thioredoxin 1 emerged as preferentially committed to phosphatidylinositol 3'-kinase/AKT pathway activation as observed in neuroblastoma. Thioredoxin 1 is a potential target for therapeutic intervention. PMID- 21641014 TI - Recent advances in the biology of Merkel cell carcinoma. AB - Recent outstanding research has rapidly revealed new aspects of the biology, etiology, and clinicopathology of Merkel cell carcinoma, a rare but highly aggressive neuroendocrine skin malignancy that affects the elderly and immunosuppressed patients. Molecular biological studies, especially the discovery of Merkel cell polyomavirus, have shed new light on the pathogenesis of the disease. Increasing evidence strongly suggests that this virus is causally related to the development of Merkel cell carcinoma. On the other hand, many studies have also indicated that a subset (approximately 20%) of Merkel cell carcinomas are not likely to be associated with the virus. Tumors with and without the virus have been shown to be significantly different in prognosis, oncogene expression, and histologic appearance, suggesting that they have different etiologies. Moreover, studies on the histopathology, immunohistochemistry, and cytogenetics have revealed several biological factors that are related to the clinical behavior and prognosis of the disease. This review summarizes the advances in the molecular biology of Merkel cell carcinoma based on recent study results. Although the exact molecular pathway of the pathogenesis of Merkel cell carcinoma remains unclear, further understanding of the pathophysiology of this tumor is expected to result in novel therapeutic approaches for management of the disease and contribute to better patient outcomes. PMID- 21641015 TI - First record of a parapithecid primate from the Oligocene of Kenya. AB - Recent excavations in northwestern Kenya have recovered a vertebrate fauna of late early or early late Oligocene age. Among the mammal remains, a fragmentary lower jaw and an isolated upper molar have been attributed to a small primate, Lokonepithecus manai gen. et sp. nov. Lokonepithecus is a primitive member of the Parapithecidae and possibly most closely related to Apidium from the Fayum. The new primate from Kenya is the youngest parapithecid known and its occurrence in the Oligocene of Kenya suggests that sub-Saharan Africa probably played a major role in the evolutionary history of several groups of mammals. PMID- 21641016 TI - Pulmonary hemorrhage associated with Henoch-Schonlein purpura in pediatric patients: case report and review of the literature. AB - OBJECTIVES: To characterize the clinical characteristics of pediatric patients with pulmonary hemorrhage and Henoch-Schonlein purpura (HSP). METHODS: Presentation of a pediatric patient with pulmonary hemorrhage associated with HSP and review of relevant cases based on a PubMed search (1966 to April 2010). RESULTS: We identified 17 previously reported pediatric cases with HSP and pulmonary hemorrhage. The most frequent clinical manifestations were rash (17 patients, 100%), proteinuria (14 patients, 82%), and abdominal pain (13 patients, 76%). Six patients (35%) had complete resolution of symptoms; 7 patients (41%) had partial recovery, and 4 patients (23%) died. Nine patients (53%) had acute respiratory failure following intubation and 3 of these patients (33%) died. Five patients were treated with methylprednisolone pulse therapy and 1 was also given low-dose cyclophosphamide treatment, but 2 of these 5 patients (40%) died. Three patients were given cyclophosphamide pulse therapy plus steroid (nonpulse or pulse) therapy, and all survived. Among the 6 nonintubated patients, all were given steroid treatment with or without an immunosuppressant drug, and all survived. In our reported case, plasma exchange appeared to help resolve the pulmonary hemorrhage and crescentic glomerulonephritis that were associated with HSP. CONCLUSIONS: For pediatric HSP patients with pulmonary hemorrhage but no respiratory failure, methylprednisolone pulse or nonpulse therapy could be the first-line therapy. In the presence of respiratory failure, cyclophosphamide pulse therapy is suggested. Plasma exchange may be considered for treatment of pulmonary renal syndrome or refractory pulmonary hemorrhage. PMID- 21641017 TI - Is there a place for cardiovascular magnetic resonance imaging in the evaluation of cardiovascular involvement in rheumatic diseases? AB - Cardiovascular magnetic resonance (CMR) is a noninvasive, nonradiating imaging technique, which provides novel information for the evaluation of cardiovascular diseases. Until now it has been successfully used for the evaluation of congenital and acquired heart diseases, cardiac tumors-mass, iron overload, and myocardial fibrosis detection. Recently, its diagnostic capabilities have been extended to the evaluation of myocardial inflammation and myocardial perfusion. Currently, it is considered the gold standard for the evaluation of volumes, mass, ejection fraction of atriums and ventricles, quantification of iron overload in different organs, detection and follow-up of myocardial inflammation, myocardial infarction and its complications, evaluation of the aorta, detection of anomalous coronary arteries, and ectatic or aneurysmatic coronary arteries. All the above applications and mainly the CMR ability to detect myocardial inflammation, perfusion defects, fibrosis, coronary and great arteries aneurysms make it a valuable tool for cardiovascular system assessment, commonly affected during the course of rheumatic diseases. The technique has been already successfully used in the evaluation of vasculitides, systemic lupus erythematosus, myositis, and scleroderma. However, further studies are needed to evaluate its usefulness as a diagnostic and monitoring tool of cardiovascular involvement in rheumatic patients. PMID- 21641018 TI - Peripheral neuropathy in patients with systemic lupus erythematosus. AB - OBJECTIVE: In patients with systemic lupus erythematosus (SLE), to determine 1) the prevalence and clinical features of peripheral neuropathies (PN) and whether they were SLE related, 2) whether there are associations between other SLE features and PN. METHODS: Patients who met the American College of Rheumatology case definition criteria for SLE peripheral neuropsychiatric syndromes were selected from the University of Toronto Lupus Clinic database. Demographic data and SLE-related clinical and laboratory data were extracted. Health-related quality of life was assessed using the mental and physical component summary score of the SF-36 questionnaire. In a nested case-control study, SLE patients with PN were matched by disease duration and compared with those without PN. RESULTS: Of 1533 patients in the database, 207 (14%) had PN. Of these, 40% were non-SLE-related. Polyneuropathy was diagnosed in 56%, mononeuritis multiplex in 9%, cranial neuropathy in 13%, and mononeuropathy in 11% of patients. Asymmetric presentation was most common (59%) and distal weakness occurred in 34%. Electrophysiologic studies indicated axonal neuropathy in 70% and signs of demyelination in 20% of patients. Compared with patients without PN, those with PN had significantly more central nervous system involvement, higher SLE-disease activity index 2000 and lower SF-36-PCS. CONCLUSIONS: The prevalence of PN is relatively high in SLE and occurs more frequently in patients with central nervous system involvement and high SLE-disease activity index. There is a predilection for asymmetric and lower extremities involvement, especially peroneal and sural nerves. This manifestation of the disease has a significant impact on the patient's quality of life. PMID- 21641019 TI - Statins in the prevention of venous thromboembolism: a meta-analysis of observational studies. AB - INTRODUCTION: Studies have established a relationship between inflammation and venous thromboembolism (VTE). Though statins modulate inflammation, it is uncertain if they prevent VTE in heterogeneous populations. A recent randomized trial demonstrated that statins prevent VTE in healthy older adults, yet this has not been well established in other groups, including younger individuals and individuals with comorbidities. The objective of this meta-analysis was to estimate the effect of statins on VTE in a heterogeneous group of adults. METHODS: We systematically reviewed the effect of statins in preventing VTE in adult inpatients and outpatients. We systematically searched MEDLINE (1966-Jan 2010), EMBASE (1980-Jan 2010), Google Scholar, Cochrane Library, PapersFirst, ProceedingsFirst, and ISI Web of Science, manually reviewed references, and contacted experts. Observational studies that compared any dose of statin to no statin or placebo, examined inpatients or outpatients, and assessed VTE, pulmonary embolism, and/or deep vein thrombosis were included. Study selection, data abstraction and study quality evaluation (using the Newcastle-Ottawa Scale) were independently conducted in duplicate. RESULTS: Four cohort studies and four case-control studies met criteria. Comparing statins to control, the odds ratio for VTE was 0.67 (95% confidence interval 0.53, 0.84), and for deep vein thrombosis was 0.53 (95% confidence interval 0.22, 1.29). The association was attenuated in lower-quality studies and studies enrolling older individuals. CONCLUSIONS: Further well-designed trials are needed to evaluate the risks and benefits of statins in preventing VTE in heterogenous populations of adults, identify high-risk subgroups, and analyze cost-effectiveness of statin use for this indication. PMID- 21641020 TI - Catheter-directed ultrasound-accelerated thrombolysis for the treatment of acute pulmonary embolism. AB - BACKGROUND: Systemic thrombolysis rapidly improves right ventricular (RV) dysfunction in patients with acute pulmonary embolism (PE) but is associated with major bleeding complications in up to 20%. The efficacy of low-dose, catheter directed ultrasound-accelerated thrombolysis (USAT) on the reversal of RV dysfunction is unknown. MATERIALS AND METHODS: We performed a retrospective analysis of 24 PE patients (60 +/- 16 years) at intermediate (n = 19) or high risk (n = 5) from the East Jefferson General Hospital who were treated with USAT (mean rt-PA dose 33.5 +/- 15.5mg over 19.7 hours) and received multiplanar contrast-enhanced chest computed tomography (CT) scans at baseline and after USAT at 38 +/- 14 hours. All CT measurements were performed by an independent core laboratory. RESULTS: The right-to-left ventricular dimension ratio (RV/LV ratio) from reconstructed CT four-chamber views at baseline of 1.33 +/- 0.24 was significantly reduced to 1.00 +/- 0.13 at follow-up by repeated-measures analysis of variance (p < 0.001). The CT-angiographic pulmonary clot burden as assessed by the modified Miller score was significantly reduced from 17.8 +/- 5.3 to 8.7 +/- 5.1 (p < 0.001). All patients were discharged alive, and there were no systemic bleeding complications but four major access site bleeding complications requiring transfusion and one suspected recurrent massive PE event. CONCLUSIONS: In patients with intermediate and high risk PE, low-dose USAT rapidly reverses right ventricular dilatation and pulmonary clot burden. PMID- 21641021 TI - Cathodal transcranial direct current stimulation of the right Wernicke's area improves comprehension in subacute stroke patients. AB - Previous studies have shown the appearance of right-sided language-related brain activity in right-handed patients after a stroke. Non-invasive brain stimulation such as transcranial direct current stimulation (tDCS) and repetitive transcranial magnetic stimulation (rTMS) have been shown to modulate excitability in the brain. Moreover, rTMS and tDCS have been found to improve naming in non fluent post-stroke aphasic patients. Here, we investigated the effect of tDCS on the comprehension of aphasic patients with subacute stroke. We hypothesized that tDCS applied to the left superior temporal gyrus (Wernicke's area) or the right Wernicke's area might be associated with recovery of comprehension ability in aphasic patients with subacute stroke. Participants included right-handed subacute stroke patients with global aphasia due to ischemic infarct of the left M1 or M2 middle cerebral artery. Patients were randomly divided into three groups: patients who received anodal tDCS applied to the left superior temporal gyrus, patients who received cathodal tDCS applied to the right superior temporal gyrus, and patients who received sham tDCS. All patients received conventional speech and language therapy during each period of tDCS application. The Korean Western Aphasia Battery (K-WAB) was used to assess all patients before and after tDCS sessions. After intervention, all patients had significant improvements in aphasia quotients, spontaneous speech, and auditory verbal comprehension. However, auditory verbal comprehension improved significantly more in patients treated with a cathode, as compared to patients in the other groups. These results are consistent with the role of Wernicke's area in language comprehension and the therapeutic effect that cathodal tDCS has on aphasia patients with subacute stroke, suggesting that tDCS may be an adjuvant treatment approach for aphasia rehabilitation therapy in patients in an early stage of stroke. PMID- 21641022 TI - Multi- and unisensory decoding of words and nonwords result in differential brain responses in dyslexic and nondyslexic adults. AB - The present functional magnetic resonance imaging (fMRI) study was designed, in order to investigate the neural substrates involved in the audiovisual processing of disyllabic German words and pseudowords. Twelve dyslexic and 13 nondyslexic adults performed a lexical decision task while stimuli were presented unimodally (either aurally or visually) or bimodally (audiovisually simultaneously). The behavioral data collected during the experiment evidenced more accurate processing for bimodally than for unimodally presented stimuli irrespective of group. Words were processed faster than pseudowords. Notably, no group differences have been found for either accuracy or for reaction times. With respect to brain responses, nondyslexic compared to dyslexic adults elicited stronger hemodynamic responses in the leftward supramarginal gyrus (SMG), as well as in the right hemispheric superior temporal sulcus (STS). Furthermore, dyslexic compared to nondyslexic adults showed reduced responses to only aurally presented signals and enhanced hemodynamic responses to audiovisual, as well as visual stimulation in the right anterior insula. Our behavioral results evidence that the two groups easily identified the two-syllabic proper nouns that we provided them with. Our fMRI results indicate that dyslexics show less neuronal involvement of heteromodal and extrasylvian regions, namely, the STS, SMG, and insula when decoding phonological information. We posit that dyslexic adults evidence deficient functioning of word processing, which could possibly be attributed to deficits in phoneme to grapheme mapping. This problem may be caused by impaired audiovisual processing in multimodal areas. PMID- 21641023 TI - Improving stable patient flow through the emergency department by utilizing evidence-based practice: one hospital's journey. AB - Many patients utilize hospital emergency services for non-emergent care. The lack of access to primary care providers, lack of insurance, and lack of information about when to use the emergency department have contributed to the crowding of emergency departments with non-emergent cases. Crowding has created longer wait times and an increased number of people leaving without a medical screening examination. Studies suggest a growing need to improve patient flow through the emergency department and to maximize resource utilization. Through a patient flow improvement project, this organization identified internal benchmarks related to total length of stay for stable patients presenting to the emergency department to promote early intervention and rapid treatment. In an attempt to meet internal as well as national benchmarks related to total length of stay, a satellite area where stable patients could be treated in a timely manner was created. One identified need was the development of a protocol that addressed the timely staffing of the satellite area to improve stable patient flow. A volume-driven protocol was developed and implemented through the use of published evidence focused on essential endpoints of measurement. The process used for the development, implementation, and evaluation of the protocol was the Iowa Model of Evidence-Based Practice. PMID- 21641024 TI - Evaluating and improving the handoff process. PMID- 21641025 TI - Needlestick and sharp object injuries among health care workers in Hamadan Province, Iran. AB - INTRODUCTION: Exposure to septic fluids through mucus and needlestick injuries is of great concern among health care workers (HCWs). The purpose of this study was to acquire epidemiological data, including occupation, level of education, and mode of exposure, in HCWs. METHODS: The data on this group was gathered during 6 months and analyzed using SPSS version 16. RESULTS: In this study, we detected 89 cases of needlestick injuries and exposure to septic body fluids in the health care centers. Nurses were the most exposed group (39.3%). The most exposed age group was 25-34 years (51.6%). Of those injured, 51.7% were evaluated for antibody titration after vaccination, and of them, 47.2% had titration above 10 mIU/mL and the remaining had titration below 10 mIU/mL. DISCUSSION: Numerous educational programs on care are still necessary to inform active workers in the health system about the warning trends and consequences in this part of Iran. PMID- 21641026 TI - Towards an improved investment approach for an effective response to HIV/AIDS. AB - Substantial changes are needed to achieve a more targeted and strategic approach to investment in the response to the HIV/AIDS epidemic that will yield long-term dividends. Until now, advocacy for resources has been done on the basis of a commodity approach that encouraged scaling up of numerous strategies in parallel, irrespective of their relative effects. We propose a strategic investment framework that is intended to support better management of national and international HIV/AIDS responses than exists with the present system. Our framework incorporates major efficiency gains through community mobilisation, synergies between programme elements, and benefits of the extension of antiretroviral therapy for prevention of HIV transmission. It proposes three categories of investment, consisting of six basic programmatic activities, interventions that create an enabling environment to achieve maximum effectiveness, and programmatic efforts in other health and development sectors related to HIV/AIDS. The yearly cost of achievement of universal access to HIV prevention, treatment, care, and support by 2015 is estimated at no less than US$22 billion. Implementation of the new investment framework would avert 12.2 million new HIV infections and 7.4 million deaths from AIDS between 2011 and 2020 compared with continuation of present approaches, and result in 29.4 million life years gained. The framework is cost effective at $1060 per life-year gained, and the additional investment proposed would be largely offset from savings in treatment costs alone. PMID- 21641027 TI - Rupture of a giant carotid-ophthalmic aneurysm. PMID- 21641028 TI - A metabonomic analysis of organ specific response to USPIO administration. AB - As ultrasmall superparamagnetic particles of iron oxides (USPIO) have been widely used in clinical medicine as MRI contrast agents, hence their potential toxicity and adverse effects following administration have attracted particular attention. In the present study, high resolution magic-angle-spinning (1)H NMR spectroscopy coupled with multivariate statistical analysis was used to directly determine the metabolic consequences of specific-tissues, including kidney, liver and spleen following the intravenous administration of USPIO. Alterations of renal, hepatic and splenic function were reflected by changes in a number of metabolic pathways including small molecules involved in energy, lipid, glucose, and amino acids metabolism. The toxicological potential and metabolic fate of USPIO seems to be linked to their surface chemistry and particle size. Hierarchical principal component analysis was used to explore the multidimensional metabolic relationships between various biological matrices such as kidney, liver, spleen, plasma and urine. Information on the involvement of USPIO in transportation, absorption, biotransformation, biodistribution and secretion was derived from metabolic correlation analysis between different organs and biofluids. Such a metabonomic strategy provides methodology for investigating the potential adverse biological effects of similar nanoparticles on the environmental and human health and assessing the drug interventions on the targeted organ. PMID- 21641029 TI - Patterning and detailed study of human hNT astrocytes on parylene-C/silicon dioxide substrates to the single cell level. AB - It is estimated that the adult human brain contains 100 billion neurons with 5-10 times as many astrocytes. Although it has been generally considered that the astrocyte is a simple supportive cell to the neuron, recent research has revealed new functionality of the astrocyte in the form of information transfer to neurons of the brain. In our previous work we developed a protocol to pattern the hNT neuron (derived from the human teratocarcinoma cell line (hNT)) on parylene C/SiO(2) substrates. In this work, we report how we have managed to pattern hNT astrocytes, on parylene-C/SiO(2) substrates to single cell resolution. This article disseminates the nanofabrication and cell culturing steps necessary for the patterning of such cells. In addition, it reports the necessary strip lengths and strip width dimensions of parylene-C that encourage high degrees of cellular coverage and single cell isolation for this cell type. The significance in patterning the hNT astrocyte on silicon chip is that it will help enable single cell and network studies into the undiscovered functionality of this interesting cell, thus, contributing to closer pathological studies of the human brain. PMID- 21641031 TI - Mechanism and consequence of chitosan-mediated reversible epithelial tight junction opening. AB - In order to increase the absorption of hydrophilic macromolecules in the small intestine, permeation enhancers such as chitosan (CS) and its derivatives have been evaluated. The aim of the current work was to investigate, on molecular levels, the effect of CS on tight junction (TJ) integrity in Caco-2 cells. The observed changes in transepithelial-electrical-resistance measurements and the staining patterns of the monolayer Caco-2 cells demonstrate that CS can transiently and reversibly open the TJs between cells, thus enhancing the paracellular permeability. TJ ultra-structures examined by transmission electron microscopy support the concept that CS did induce transient opening of TJs. We then assessed TJ disruption at the gene and protein expression levels. Our data indicate that exposure to CS followed by recovery resulted in a significant increase in claudin-4 (Cldn4) gene transcription. Additionally, CS treatment induced redistribution of the TJ protein CLDN4 intracellularly following by its degradation in lysosomes, which represented an important contributing factor in TJ weakening, leading to the opening of TJs. The recovery of TJ after CS disruption required CLDN4 protein synthesis. These results suggest that CS regulates TJs by inducing changes in transmembrane CLDN4 protein. Understanding the mechanism of interaction between CS and epithelial cells is of paramount importance and needs to be established to aid further development in the use of CS to mediate the trans-epithelial drug delivery. PMID- 21641030 TI - Biodegradation, biocompatibility, and drug delivery in poly(omega pentadecalactone-co-p-dioxanone) copolyesters. AB - Poly(omega-pentadecalactone-co-p-dioxanone) [poly(PDL-co-DO)] copolyesters are copolymers of an isodimorphic system, which remain semicrystalline over the whole range of compositions. Here, we evaluated enzymatically synthesized poly(PDL-co DO) copolymers as new materials for biomedical applications. In vivo experiments using mice, showed that the copolyesters are well tolerated, with tissue responses that are comparable to poly(p-dioxanone). In addition, the copolymers were found to degrade hydrolytically at controlled rates over a period of several months under physiological conditions. The poly(PDL-co-DO) copolymers with up to 69 mol% DO units were successfully transformed to free-standing nanoparticles that are capable of encapsulating an anticancer drug, doxorubicin, or a polynucleotide, siRNA. Drug- or siRNA-loaded nanoparticles exhibited controlled and continuous release of agent over many weeks. In addition, siLUC-encapsulated poly(PDL-co-DO) nanoparticles were active in inhibiting luciferase gene expression in LUC-RKO cells. Because of substantial differences in structure and hydrophobicity between PDL and DO units, poly(PDL-co-DO) biodegradation rate and physical properties can be tuned over a wide range depending on the copolymer composition. Our results demonstrate that the semicrystalline and biodegradable poly(PDL-co-DO) copolyesters are promising biomaterials to serve as drug carriers, as well as potential raw materials for constructing bioabsorbable sutures and other medical devices. PMID- 21641032 TI - Association between birth order and immunological parameters? PMID- 21641033 TI - Investigation of the role of sigma1-receptors in inositol 1,4,5-trisphosphate dependent calcium signaling in hepatocytes. AB - In hepatocytes, as in other cell types, Ca(2+) signaling is subject to complex regulations, which result largely from the intrinsic characteristics of the different inositol 1,4,5-trisphosphate receptor (InsP(3)R) isoforms and from their interactions with other proteins. Although sigma1 receptors (Sig-1Rs) are widely expressed in the liver, their involvement in hepatic Ca(2+) signaling remains unknown. We here report that in this cell type Sig-1R interact with type 1 isoforms of the InsP(3) receptors (InsP(3)R-1). These results obtained by immunoprecipitation experiments are confirmed by the observation that Sig-1R proteins and InsP(3)R-1 colocalize in hepatocytes. However, Sig-1R ligands have no effect on InsP(3)-induced Ca(2+) release in hepatocytes. This can be explained by the rather low expression level expression of InsP(3)R-1. In contrast, we find that Sig-1R ligands can inhibit agonist-induced Ca(2+) signaling via an inhibitory effect on InsP(3) synthesis. We show that this inhibition is due to the stimulation of PKC activity by Sig-1R, resulting in the well-known down regulation of the signaling pathway responsible for the transduction of the extracellular stimulus into InsP(3) synthesis. The PKC sensitive to Sig-1R activity belongs to the family of conventional PKC, but the precise molecular mechanism of this regulation remains to be elucidated. PMID- 21641034 TI - Chronic high cytosolic calcium decreases AICAR-induced AMPK activity via calcium/calmodulin activated protein kinase II signaling cascade. AB - Calcium is important for muscle contraction and controls many cellular processes. Although there is evidence that calcium-mediated signals regulate AMP-activated protein kinase (AMPK) activity, the molecular mechanisms by which calcium regulates AMPK are poorly understood. To compare the function of sustained vs. intermittent calcium oscillations on AMPK activity and define specific signals in this pathway, we administered mice with aminoimidazole-carboxamide-ribonucleotide (AICAR) and caffeine with or without dantrolene. AMPK activity was increased by 10 d AICAR treatment (P < 0.01). Ten day caffeine treatment decreased AICAR induced AMPK activity to control level. This repressed AMPK activity was blocked by dantrolene. Different calcium frequencies were simulated in C2C12 myotubes by alternating media containing caffeine and dantrolene. Intermittent calcium oscillation increased AMPK activity compared to control (P < 0.05), whereas sustained calcium oscillation decreases AICAR-induced AMPK activity to control level. This result suggests a biphasic control of AMPK activity by calcium. Knockdown of CaMKII expression by short-hairpin RNA resulted in increased AMPK phosphorylation by AICAR even in the presence of caffeine. These data show different calcium oscillations elicit distinct responses in muscle cells suggesting that the negative effects of chronic calcium treatment on AMPK activity is partly mediated through the CaMKII signals. PMID- 21641035 TI - Chromosomal translocation t(1;9)(q24;q34) in acute lymphoblastic leukemia patient involving the ABL1 gene. PMID- 21641036 TI - Agranulocytosis in cocaine users in Ohio: suspected levamisole taint. PMID- 21641037 TI - Molecular response to first line nilotinib in a patient with e19a2 BCR-ABL1 chronic myeloid leukemia. PMID- 21641038 TI - Sustained trilineage hematopoietic recovery in a patient with refractory anemia, del(13)(q12q22), and paroxysmal nocturnal hemoglobinuria-type cells treated with immunosuppressive therapy. PMID- 21641039 TI - Transgenic overexpression of toxin-related ecto-ADP-ribosyltransferase ART2.2 sensitizes T cells but not B cells to NAD-induced cell death. AB - T cells constitutively express low amounts of a toxin-related ADP-ribosylating ecto-enzyme, ART2.2. In inflammatory settings, cells release NAD, the substrate for ART2.2. The ART2.2 catalyzed ADP-ribosylation of cell surface proteins induces cell death. However, the low expression levels of ART2.2 have hampered analysis of ART2.2 in physiological settings. Here we report the generation of transgenic mice over-expressing ART2.2 under the control of the H2K promoter and IgMU enhancer. ART2.2 transgenic mice were healthy and fertile and exhibited normal development of the major lymphocyte subsets. Most T cells and a small subpopulation of B cells from transgenic mice showed more than 10-fold higher levels of ART2.2 expression than their wild-type counterparts. Exposure of ART2.2 transgenic T cells to low, submicromolar concentrations of NAD caused cell membrane alterations including uptake of propidium iodide, externalization of phosphatidylserine, and shedding of CD62L, while ART2.2-transgenic B cells were resistant to NAD. The ART2.2-overexpressing animals described here confirm that ART2.2 is an essential component for the regulation of T-cell functions by extracellular NAD and provide a useful tool to further elucidate the function of ART2.2 in vivo. PMID- 21641040 TI - Macular pigment changes in pseudophakic eyes quantified with resonance Raman spectroscopy. AB - PURPOSE: We examined changes in macular pigment optical density (MPOD) levels after cataract surgery and compared the MPOD between eyes with clear intraocular lenses (IOLs) and yellow-tinted IOLs. DESIGN: Prospective, comparative case series. PARTICIPANTS: The MPOD levels were measured in 480 eyes of 337 patients after cataract surgery. Among them, the data from 259 eyes (clear IOL group, 121 eyes; yellow-tinted IOL group, 138 eyes) of 259 Japanese patients were selected for statistical analyses on the basis of the inclusion criteria: a postoperative visual acuity (VA) of >=0.8 and no fundus diseases. Only 1 eye of each patient was enrolled. Patients provided informed consent to participate in this study on the basis of the approval of the institutional review board before surgery. METHODS: The patients selected the type of IOL to be implanted. The MPOD levels were measured using resonance Raman spectroscopy on day 1 (baseline value); months 1, 3, and 6; and years 1 and 2 postoperatively. MAIN OUTCOME MEASURES: The difference in MPOD levels between the IOL groups was analyzed by unpaired t tests. The following parameters were analyzed by multiple regression analysis: age, gender, body mass index (BMI), smoking history, glaucoma, diabetes, preoperative VA, preoperative refractive error, and IOL power and type. RESULTS: We found no significant differences in the baseline characteristics between the 2 groups. Until 6 months postoperatively, the MPOD levels did not differ significantly between the groups. However, from 1 year onward, the levels were significantly higher in the yellow-tinted IOL group compared with the clear IOL group. By multiple regression analysis, 1 day postoperatively, older age and diabetes were correlated with lower MPOD levels; 1 year postoperatively and thereafter, however, lower MPOD levels were correlated with clear IOLs. CONCLUSIONS: Cataract surgery with clear IOLs induced a greater decrease in macular pigment levels compared with yellow-tinted IOLs during a longer follow-up period. These findings agreed with observations that excessive light exposure is associated inversely with MPOD, because clear IOLs transmit higher intensities of blue light than yellow-tinted IOLs. PMID- 21641041 TI - Epidemiology of syphilis, gonorrhea and chlamydia in Romania, 1980-2009. AB - OBJECTIVES: This study aims to analyze the epidemiology of syphilis, gonorrhea and genital chlamydial infections in Romania during 1980-2009 when radical changes occurred during transition from a communist regime to a democratic society. METHODS: Surveillance data was retrieved from the National Center for Surveillance and Control of the Communicable Disease in Romania and the World Health Organization. RESULTS: The mean annual incidences of syphilis, gonorrhea and genital chlamydia infections in Romania were 25.2 +/- 15 cases per 100,000 inhabitants (1980-2009), 33.2 +/- 26.6/100,000 (1980-2009) and 0.3 +/- 0.4/100,000 (1994-2009), respectively. Although the infection rates for syphilis followed a significant ascending trend over the study period (p < 0.001), they declined dramatically since 2002 (p < 0.001). There was a significant decrease in gonorrhea-affected persons over the study period (p < 0.0001), while genital chlamydia infections registered a general ascending trend (p = 0.02) with insignificant decrease since 2006. CONCLUSIONS: The declining trend of syphilis and gonorrhea infections in the general population especially during the last years is very encouraging but should not challenge the amount of resources dedicated to sexually transmitted infections. Consequently, compelling strategies and policies based mainly on education of the population should be put forth in order to advance efficient prevention. PMID- 21641042 TI - Increasing incidence of iliopsoas abscesses with MRSA as a predominant pathogen. AB - OBJECTIVES: Iliopsoas abscesses (IPAs) are rare infections involving purulence within the muscles of the iliopsoas compartment, seldom due to Methicillin resistant Staphylococcus aureus (MRSA) historically. This study was designed to evaluate the clinical characteristics and outcomes of patients with IPAs. METHODS: A retrospective analysis of the demographics, clinical presentation, microbiologic data and treatment modalities among patients with IPAs from 1993 to 2007 at The Johns Hopkins Hospital was performed. RESULTS: Among 61 patients with IPAs, infection rates increased from 0.5 cases/10,000 admissions (1993-2004) to 6.5 cases/10,000 admissions (2005-2007) (P < 0.001). An adjacent infectious focus was identified in 80% of patients, from skeletal (48%), intra-abdominal (23%), vascular (5%), genitourinary (3%), and cutaneous sources (2%). During 2005-2007, MRSA became a predominant pathogen, accounting for 25% of all cases and 37% of cases with a definitive microbiologic diagnosis (P = 0.006). Patients with IPAs >2 cm were more likely to undergo drainage, with trends toward longer hospitalizations, longer antibiotic courses, and increased odds of securing a definitive microbiologic diagnosis. CONCLUSIONS: Since 2005, rates of IPA have dramatically increased, with MRSA now the leading cause of infection. Knowledge of common pathogens should guide antimicrobial therapy including empiric coverage for MRSA in institutions with similar populations, especially if culture data are not available. PMID- 21641043 TI - Reactivity of affect and self-esteem during remission in bipolar affective disorder: an experimental investigation. AB - BACKGROUND: Bipolar affective disorder (BPAD) is characterised by a lifelong vulnerability to develop episodes of depressed or elevated mood in response to stressful life events involving achievement or failure. We hypothesised that this latent vulnerability can manifest as reactivity of affect and self-esteem to experimentally induced experiences of success and failure and is shaped by history of childhood adversity. METHODS: Twenty-four people with remitted BPAD and twenty-four healthy controls underwent anagram-solving tasks designed to generate experiences of success and failure in two separate sessions. Positive and negative affect and implicit and explicit self-esteem were measured before and after each task. Early adversity was measured by Childhood Trauma Questionnaire. RESULTS: People with BPAD showed larger reactivity of affect and explicit self-esteem in response to experimental success and failure than did healthy controls. There were no significant differences in reactivity of implicit self-esteem. History of childhood trauma predicted increased affective reactivity to failure but not to success. LIMITATIONS: We used a convenience sample. CONCLUSIONS: The present experimental paradigm reveals reactivity of affect and self-esteem as features of BPAD, which are present even during good remission and thus are accessible as targets of interventions aiming at relapse prevention. Differential associations with childhood adversity indicate aetiological heterogeneity, with reactivity to failure influenced by early trauma and reactivity to success driven by other mechanisms. PMID- 21641044 TI - Val1483Ile polymorphism in the fatty acid synthase gene was associated with depressive symptoms under the influence of psychological stress. AB - BACKGROUND: To study the association between lipid-metabolism and depressive symptoms, genetic polymorphisms in serotonin transporter linked promoter region (5-HTTLPR) and fatty acid synthase gene (FASN) were investigated. METHOD: A cross sectional study was conducted on 177 women (n = 166) and men (n = 15) recruited from workers in a hospital and nursing homes in Japan. Depressive symptoms were assessed by the Center for Epidemiologic Studies Depression (CES-D) scale and perceived psychological stress was measured using visual analogue scale (VAS). The genotypes of 5-HTTLPR (insertion/deletion; L/S), and FASN (Val1483Ile) were determined by the PCR methods. Linear regression analysis was performed, in which CES-D scores served as a dependent variable, and VAS scores, gene polymorphism, and confounders as independent variables. RESULTS: Under the influence of perceived stress, S/S carriers of the 5-HTTLPR gene showed significantly higher CES-D scores in comparison with L/L+L/S carriers (F = 8.2, standardised beta = 0.15, p < 0.05). Regression analysis also confirmed that CES-D scores in participants with Ile/Ile+Val/Ile genotypes of the FASN gene were significantly higher than those with Val/Val genotype (F = 8.4, standardised beta = 0.16, p<0.05). In relation to physical features, BMI among participants with S/S genotype of 5-HTTLPR was significantly lower compared with those with L/L+L/S genotypes. CONCLUSIONS: The Val1483Ile polymorphism in the FASN was associated with depressive symptoms under the influence of psychological stress. The S variant of 5-HTTLPR was related with less obese. PMID- 21641045 TI - The role of affective temperaments in metabolic control in patients with type 2 diabetes. AB - BACKGROUND: In non-diabetic populations, anxious and depressive affective temperaments are associated with stress and distress disorders (anxious and depressive). In type 2 diabetes, distress with diabetes and distress disorders have been associated with metabolic control. There are few studies undertaken on temperament and metabolic control in patients with type 2 diabetes. The aim of our study was to examine the independent association between affective temperaments and metabolic control in patients with type 2 diabetes. METHODS: The sample included 90 patients with type 2 diabetes from two outpatient clinics aged 53.54 (SD +/- 8.05) years and with 66.7% female gender. Depression was evaluated by using the MADRS observer rating scale (mean value: 16.38 +/- 11). RESULTS: Patients with excessive depressive and anxious temperaments had more depressive symptoms, worse psychological adjustment to diabetes and worse metabolic control. In logistic regression (crude model), only depressive temperament was independently associated with metabolic control. CONCLUSIONS: Affective temperaments are associated with psychological adjustment to diabetes and metabolic control in patients with type 2 diabetes. Patients above threshold on depressive temperament should be considered for greater scrutiny and psycho education by the diabetes clinic staff. PMID- 21641046 TI - The validity of DSM symptoms for depression and anxiety disorders during pregnancy. AB - BACKGROUND: The applicability of the symptom criteria for diagnostic mood and anxiety disorders has in recent times been questioned for women in the perinatal period, due to the overlap of diagnostic symptoms with normal symptoms due to the physical changes of pregnancy or postpartum, (e.g., sleep difficulties). METHOD: 118 women in their second or early third trimester of pregnancy participated in a telephone interview that included the depression and anxiety modules of the MINI diagnostic interview, and an attributional probe question asking the woman whether endorsement of a symptom was due to the physical changes of her pregnancy or due to her mood or worries. RESULTS: 66% of pregnant women who met criteria for major depression reported that a sufficient number of their symptoms were due to the normal physical changes of their pregnancy, such that they would no longer meet criteria for a diagnosis of major depression. Attributional probing resulted in the rate of major depression falling from 6.8% to 1.7%. The impact on anxiety disorders appears to be less. LIMITATIONS: An important issue is whether women's attributional perceptions are valid, though the face validity of some of their responses means that credence should be given to the findings. In addition, it would be useful from a validity perspective to undertake the same attributional probing with other populations where symptom presence is most likely due to mood, such as unemployed or recently separated adults without physical complaints. CONCLUSION: Rates of depressive disorders in pregnancy, using DSM symptom criteria, are significantly over-estimated due to the natural occurrence of many of the symptoms as a result of the pregnancy. Rates of anxiety disorders are also inflated, but to a lesser degree. This means that the validation of self-report mood measures, typically done against DSM diagnoses, is likely to have produced erroneous findings (e.g., optimum cut-off scores). It is probable that a similar finding would be obtained for the postpartum period. Thus future use of DSM symptom criteria for depression and anxiety in perinatal mental health work should use some form of attributional probing question to more accurately understand the applicability of symptoms to a diagnosis in this population. PMID- 21641047 TI - Fronto-extracephalic transcranial direct current stimulation as a treatment for major depression: an open-label pilot study. AB - BACKGROUND: Several recent trials have reported transcranial direct current stimulation (tDCS) to be effective in treating depression, though the relative benefits of different electrode montages remain unexplored. Whereas all recent studies have used a bifrontal (BF) electrode montage, studies published in the 1960s and 1970s placed one electrode in an extracephalic position, with some positive reports of efficacy. This study investigated the efficacy and safety of tDCS given with a fronto-extracephalic (F-EX) montage. METHODS: 2 mA tDCS was administered for 20 min every weekday over four weeks in 11 participants with a Major Depressive Episode who had previously shown inadequate response to, or relapsed following, a course of BF tDCS. For F-EX tDCS the anode was placed on the left dorsolateral prefrontal cortex and the cathode on the right upper arm. Depression severity and neuropsychological function were assessed before and after the treatment course. Antidepressant response was compared across an equivalent treatment period for both montages. RESULTS: F-EX tDCS was shown to be safe and well tolerated. Depression ratings improved after acute treatment on the Montgomery Asberg Depression Rating Scale (p < 0.001). Participants showed greater initial treatment response with F-EX tDCS than with BF tDCS (p < 0.001). LIMITATIONS: This was an open label pilot study. The two comparison treatments were applied consecutively. CONCLUSION: F-EX tDCS appears to be safe and to have antidepressant effects, and may lead to more rapid improvement than tDCS given with a BF montage. PMID- 21641048 TI - Excessive Internet gaming and decision making: do excessive World of Warcraft players have problems in decision making under risky conditions? AB - The dysfunctional behavior of excessive Internet gamers, such as preferring the immediate reward (to play World of Warcraft) despite the negative long-term consequences may be comparable with the dysfunctional behavior in substance abusers or individuals with behavioral addictions, e.g. pathological gambling. In these disorders, general decision-making deficits have been demonstrated. Hence, the aim of the present work was to examine decision-making competences of excessive World of Warcraft players. Nineteen excessive Internet gamers (EIG) and a control group (CG) consisting of 19 non-gamers were compared with respect to decision-making abilities. The Game of Dice Task (GDT) was applied to measure decision-making under risky conditions. Furthermore psychological-psychiatric symptoms were assessed in both groups. The EIG showed a reduced decision-making ability in the GDT. Furthermore the EIG group showed a higher psychological psychiatric symptomatology in contrast to the CG. The results indicate that the reduced decision-making ability of EIG is comparable with patients with other forms of behavioral addiction (e.g. pathological gambling), impulse control disorders or substance abusers. Thus, these results suggest that excessive Internet gaming may be based on a myopia for the future, meaning that EIG prefer to play World of Warcraft despite the negative long-term consequences in social or work domains of life. PMID- 21641049 TI - A mimotope peptide of Abeta42 fibril-specific antibodies with Abeta42 fibrillation inhibitory activity induces anti-Abeta42 conformer antibody response by a displayed form on an M13 phage in mice. AB - In Alzheimer's disease (AD), amyloid-beta (Abeta) peptides accumulate in the brain in different forms, including fibrils and oligomers. Recently, we established three distinct conformation-dependent human single-chain Fv (scFv) antibodies, including B6 scFv, which bound to Abeta42 fibril but not to soluble form Abeta, inhibiting Abeta42 fibril formation. In this study, we determined the mimotopes of these antibodies and found a common mimotope sequence, B6-C15, using the Ph.D.-C7C phage library. The B6-C15 showed weak homology to the C-terminus of Abeta42 containing GXXXG dimerization motifs. We synthesized the peptide of B6 C15 fused with biotinylated TAT at the N-terminus (TAT-B6-C15) and characterized its biochemical features on an Abeta42-fibrillation reaction in vitro. We demonstrated that, first, TAT-B6-C15 inhibited Abeta42 fibril formation; secondly, TAT-B6-C15 bound to prefibril Abeta42 oligomers but not to monomers, trimers, tetramers, fibrils, or ultrasonicated fragments; thirdly, TAT-B6-C15 inhibited Abeta42-induced cytotoxicity against human SH-SY5Y neuroblastoma cells; and, fourthly, when mice were administered B6-C15-phages dissolved in phosphate buffered saline, the anti-Abeta42 conformer IgG antibody response was induced. These results suggested that the B6-C15 peptide might provide unique opportunities to analyze the Abeta42 fibrillation pathway and develop a vaccine vehicle for Alzheimer's disease. PMID- 21641050 TI - The strength of age effect on tooth loss and periodontal condition in Korean elderly. AB - Aging is a well-known risk factor associated with oral diseases. The aim of this cross-sectional study was to compare tooth loss and periodontal health between the relatively young elderly (65-74 years) and the old elderly (>= 75 years) and to investigate the strength of the age effect on oral health status in the Korean elderly. Study population 65 years of age or older were selected from the participants of the Korean National Oral Health Survey (2006) (n = 1193). Oral examination was conducted by eight dentists trained in the World Health Organization (WHO) recommended examination procedure. The chi-square test, multiple regression analyses and multinomial logistic regression analyses were performed using SAS 9.1.3. The oral health status including decayed, missing, and filled teeth (DMFT), missing teeth, and residual teeth significantly differed between the young elderly and the old elderly (p < 0.01). Moreover, the regression coefficients of tooth loss linearly increased across different age groups (5-year intervals, starting at age 65 years) (p < 0.05). However, the odds ratios of periodontal health did not significantly differ across 5-year interval age groups. The findings that age and the number of missing teeth are significantly and linearly related could contribute to the development of oral health care and promotion programs for the elderly tailored to their own age. PMID- 21641051 TI - Galectin-3 in heart failure: high levels are associated with all-cause mortality. PMID- 21641052 TI - A call for adult congenital heart disease patient participation in cardiac rehabilitation. PMID- 21641053 TI - Evaluation of neointimal coverage and apposition with various drug-eluting stents over 12 months after implantation by optical coherence tomography. AB - BACKGROUND: Optical coherence tomography (OCT) is an imaging technology with high resolution which provides new opportunities for evaluating the vascular healing reaction after stent implantation. We used OCT to compare neointimal coverage and stent strut apposition with different types of drug-eluting stent (DES) at more than 12 months follow-up. METHODS: Fifty five patients who accepted 72 DESs (19 sirolimus-eluting stents [SES, Cypher Select], 27 paclitaxel-eluting stents [PES, Taxus Liberte], 26 zotarolimus-eluting stents [ZES, Endeavor]) were enrolled in our study. OCT was performed at 12-18 months after stent implantation. The neointimal hyperplasia (NIH) thickness and stent strut apposition at 1-mm interval and the presence of thrombus in each stent were observed. RESULTS: The average NIH thickness (0.1806 +/- 0.1726 mm in SES vs. 0.2558 +/- 0.2187 mm in PES vs. 0.2983 +/- 0.2567 mm in ZES, p<0.001) and percentage of NIH area (17.21 +/- 14.05% in SES vs. 23.09 +/- 14.53% in PES vs. 28.73 +/- 20.47% in ZES, p=0.002) were significantly greater in ZES than in other DESs. The prevalence of uncovered struts (8.96% in SES vs. 6.19% in PES vs. 2.98% in ZES, p<0.01) and malapposed struts (6.36% in SES vs. 2.32% in PES vs. 1.69% in ZES, p<0.01) was significantly lower in ZES than in other DESs. Thrombus was less frequently observed in ZES than in other DESs (SES 1.70% vs. PES 0.83% vs. ZES 0.17%, p<0.01). CONCLUSIONS: Compared with SES and PES, ZES showed lower prevalence of uncovered struts and malapposed struts at more than 12 months follow-up. PMID- 21641054 TI - Visceral abdominal adipose tissue and coronary atherosclerosis in asymptomatic diabetics. AB - BACKGROUND: Visceral abdominal adipose tissue (VAT) may play an active role in the progression of coronary atherosclerosis. We examined the relation between VAT, non-alcoholic fatty liver disease and extent of coronary atheroma in patients with type 2 diabetes mellitus but no known coronary artery disease. METHODS: Coronary artery calcium and area, distribution and thickness of upper abdominal fat were measured in selected axial cross-sections from non-enhanced computed tomography (CT) scans of the chest. Coronary atheroma was assessed visually on a per vessel basis from 64 slice CT angiography using axial views and multi-format reconstructions. Fatty liver was diagnosed when liver density was <40 Hounsfield units (HU) or >=10 HU below spleen density. RESULTS: The area of VAT was increased in patients with versus without multi-vessel coronary artery plaque (237.0 +/- 101.4 vs 179.2 +/- 79.4 mm(2), p<0.001). Waist circumference (101.6 +/- 12.3 versus 95.3 +/- 13.8 cm) and internal abdominal diameter (218.7 +/- 33.0 vs 194.6 +/- 25.7 mm) (both p<0.001) were increased in patients with multi-vessel plaque whereas subcutaneous fat was unrelated to coronary plaque. Presence of fatty liver (93/318 patients, 29.2%) did not correlate with presence or extent of coronary plaque. The correlation of VAT with multi-vessel plaque although nominally independent of the metabolic syndrome (p=0.04) was not independent of waist circumference. CONCLUSION: In asymptomatic subjects with DM and no history of CAD area of VAT correlated with the presence and extent of coronary atheroma but as a risk predictor added little independent information to that obtained by more readily obtainable measures of adiposity-waist circumference and internal abdominal diameter. PMID- 21641055 TI - Electromechanical left atrial disturbances in acute inferior myocardial infarction: an evidence for ischaemic dysfunction. PMID- 21641056 TI - Intracoronary stem cell injection improves left ventricular remodeling after acute myocardial infarction: an updated meta-analysis of randomized trials. PMID- 21641057 TI - Urinary retention and heart failure: one of the many precipitating causes of decompensation. PMID- 21641058 TI - Association between cardiopulmonary exercise and dobutamine stress testing in ambulatory patients with idiopathic dilated cardiomyopathy: a comparison with peak VO2 and VE/VCO2 slope. AB - BACKGROUND: Both peak VO(2) and VE/VCO(2) slope are considered to be useful predictors of cardiovascular events. The left ventricular (LV) response to dobutamine stress testing (DST) also provides useful prognostic information. However, the relationship between these variables has not been fully investigated. Therefore, the aim of this study is to investigate the association between myocardial contractile reserve measured by DST and cardiopulmonary exercise testing (CPX) variables in patients with idiopathic dilated cardiomyopathy (IDCM). METHODS: Thirty-eight patients were subjected to CPX as well as cardiac catheterization for measurement of LV pressure. The maximum first derivative of LV pressure (LV dP/dt(max)) was measured at baseline and during dobutamine infusion at incremental doses of 5, 10, and 15 MUg kg(-1)min(-1). LV dP/dt(max) at baseline and the percentage increase in LV dP/dt(max) (DeltaLV dP/dt(max)) induced by DST served as indices of LV contractility and myocardial contractile reserve, respectively. RESULTS: Peak VO(2), and VE/VCO(2) slope were 18.6 mL kg(-1)min(-1) and 32.3, respectively. Peak VO(2) was not correlated with LV dP/dt(max) at baseline. However, peak VO(2) was significantly correlated with DeltaLV dP/dt(max), and the correlation became more pronounced as the dose of dobutamine was increased. There was no correlation between VE/VCO(2) slope and DeltaLV dP/dt(max). Multivariate regression analysis revealed that DeltaLV dP/dt(max) was independently correlated with peak VO(2) (p=0.011). CONCLUSIONS: Peak VO(2), but not VE/VCO(2) slope, may reflect myocardial contractile reserve in ambulatory patients with IDCM. This study population is small, and therefore large confirmatory studies are needed. PMID- 21641059 TI - Mortality by cerebro-/cardio-vascular disease among middle-aged residents in Japan. PMID- 21641060 TI - Current pathophysiological concepts and management of pulmonary hypertension. AB - Pulmonary hypertension (PH), increasingly recognized as a major health burden, remains underdiagnosed due mainly to the unspecific symptoms. Pulmonary arterial hypertension (PAH) has been extensively investigated. Pathophysiological knowledge derives mostly from experimental models. Paradoxically, common non-PAH PH forms remain largely unexplored. Drugs targeting lung vascular tonus became available during the last two decades, notwithstanding the disease progresses in many patients. The aim of this review is to summarize recent advances in epidemiology, pathophysiology and management with particular focus on associated myocardial and systemic compromise and experimental therapeutic possibilities. PAH, currently viewed as a panvasculopathy, is due to a crosstalk between endothelial and smooth muscle cells, inflammatory activation and altered subcellular pathways. Cardiac cachexia and right ventricular compromise are fundamental determinants of PH prognosis. Combined vasodilator therapy is already mainstay for refractory cases, but drugs directed at these new pathophysiological pathways may constitute a significant advance. PMID- 21641061 TI - Development of an easily applicable risk score model for contrast-induced nephropathy prediction after percutaneous coronary intervention: a novel approach tailored to current practice. AB - BACKGROUND: Several risk factors for contrast-induced nephropathy (CIN) after percutaneous coronary intervention (PCI) have been identified. The cumulative effect of these risk factors on renal function has been assessed with the development of risk score models in a number of studies. However, concerns were raised that estimates of the risk attributable to individual factors may be unreliable. We sought to develop a simple risk score for developing CIN after PCI irrespective of use of prophylactic measures and also capturing the effect of pre intervention medication and presence of various co-morbidities. METHODS: Consecutive patients treated with elective or urgent PCI at our cardiac catheterization laboratory were enrolled (derivation cohort n = 488, validation cohort n = 200). CIN was defined as increase >= 25% and/or >= 0.5 mg/dl in serum creatinine at 48 h after PCI vs baseline. Multivariable logistic regression analysis was then performed to identify independent predictors of CIN (pre existing renal disease, metformin use, history of previous PCI, peripheral arterial disease and >= 300 ml of contrast volume). RESULTS: The incidence of CIN in the development cohort was 10.2% with a significant trend across increasing score values (p < 0.001). The model demonstrated good discriminating power (c statistic 0.759) and excellent calibration (calibration slope 0.91). The model was validated internally by bootstrapping in 1000 samples (c-statistic 0.753) and in a cohort of 200 patients (c-statistic 0.864) demonstrating stable performance. CONCLUSIONS: The proposed risk score is easily applicable and allows for practically simple risk assessment compared to other published scores while at the same time overcomes drawbacks of previous model designs. PMID- 21641062 TI - Time is money: outpatient waiting times and health insurance choices of elderly veterans in the United States. AB - Growth in the number of days between an appointment request and the actual appointment reduces demand. Although such waiting times are relatively low in the US, current policy initiatives could cause them to increase. We estimate multiple equation models of physician utilization and insurance plan choice for Medicare eligible veterans. We find that a 10% increase in VA waiting times increases demand for Medigap insurance by 5%, implying that a representative patient would be indifferent between waiting an average of 5 more days for VA appointments and paying $300 more in annual premium. PMID- 21641063 TI - Income and the utilization of long-term care services: evidence from the Social Security benefit notch. AB - This paper estimates the impact of income on the long-term care utilization of elderly Americans using a natural experiment that led otherwise similar retirees to receive significantly different Social Security payments based on their year of birth. Using data from the 1993 and 1995 waves of the AHEAD, we estimate instrumental variables models and find that a positive permanent income shock lowers nursing home use but increases the utilization of paid home care services. We find some suggestive evidence that the effects are due to substitution of home care for nursing home utilization. The magnitude of these estimates suggests that moderate reductions in post-retirement income would significantly alter long-term utilization patterns among elderly individuals. PMID- 21641064 TI - Stereotactic body radiotherapy (SBRT) for oligometastatic lung tumors from colorectal cancer and other primary cancers in comparison with primary lung cancer. AB - PURPOSE: To analyze local control of oligometastatic lung tumors (OLTs) compared with that of primary lung cancer after stereotactic body radiotherapy (SBRT). MATERIALS AND METHODS: Retrospective record review of patients with OLTs who received SBRT with 50Gy in 5 fractions. Local control rates (LCRs), toxicities, and factors of prognostic significance were assessed. RESULTS: Twenty-one colorectal OLTs, 23 OLTs from other origins, and 188 primary lung cancers were included. Multivariate analysis revealed only tumor origin was prognostically significant (p<0.05). The 1-year/2-year LCRs in colorectal OLTs and OLTs from other origins were 80%/72% and 94%/94%, respectively. The LCR in colorectal OLTs was significantly worse than that in OLTs from the other origins and primary lung cancers with pathological and clinical diagnosis (p<0.05, p<0.0001 and p<0.005). Among 44 OLT patients, Grades 2 and 3 radiation pneumonitis were identified in 2 and 1 patients, respectively. No other toxicities of more than Grade 3 occurred. CONCLUSION: SBRT for OLTs is tolerable. The LCR for OLTs from origins other than colorectal cancer is excellent. However, LCR for colorectal OLTs is worse than that from other origins. Therefore dose escalation should be considered to achieve good local control for colorectal OLTs. PMID- 21641065 TI - The future development of radiation oncology in Europe: a personal view. PMID- 21641066 TI - Radiation therapy of post-mastectomy patients with positive nodes using fixed beam tomotherapy. AB - PURPOSES: To develop an optimized dosimetric class solution for post-mastectomy irradiation for fixed beam tomotherapy (FBT). METHODS AND MATERIALS: CT simulation scans from 10 post-mastectomy patients were used to generate plans with planning target volumes (PTVs) that included the chest wall, axillary nodes and supraclavical nodes using FBT and helical tomotherapy (HT) with 3D and IMRT modes and the resultant dosimetry was compared to conventional IMRT. FBT IMRT plans were analyzed with both 4 (4FBT) and 11-field (11FBT) plans. Important organs at risk (OARs) included the heart, ipsilateral and contralateral lungs, esophagus and contralateral breast. In all plans, the spinal cord and contralateral lung were completely blocked while the heart and ipsilateral lung were directionally blocked. Doses to the contralateral breast were minimized. Each plan was evaluated for its delivery time, percentile volume of lung receiving x Gy (V(x)) and dose received by x percent volume (D(x)). D(1) and D(5) were used to measure the maximum dose to the OAR or PTV, D(95) and D(99) were used to measure the minimal dose to the PTV. RESULTS: Compared to the conventional IMRT technique, HT IMRT, 11FBT IMRT and 11FBT 3D significantly reduced the D(1) of the heart in cases of left-sided tumors by 13%, 41% and 36%, and the V(10) of ipsilateral lung in all cases by 26%, 49% and 46%, respectively. A close to 90% reduction in the contralateral breast dose was also observed with the 11FBT plans. Target dose homogeneity of 11FBT 3D plans is inferior to that of the HT and conventional IMRT plans but the treatment delivery time, 7.59 min, was significantly shorter by 3 min. 4FBT IMRT resulted in clinically unacceptable heterogeneity with high dose regions in both the PTV and normal tissue. CONCLUSIONS: A class solution based on an 11 beam configuration was established to optimize the dosimetry of fixed beam tomotherapy planning for post-mastectomy patients. The 11FBT plans were deliverable in clinically efficient treatment times. PMID- 21641067 TI - A pooled analysis of arc-based image-guided simultaneous integrated boost radiation therapy for oligometastatic brain metastases. AB - PURPOSE: To report pooled overall survival and time to radiological intracranial progression results related to arc-based image-guided radiotherapy for dose escalation of oligometastatic disease of the brain. METHODS AND MATERIALS: Anonymized patient, tumor, and treatment data were pooled from the VU University medical center (VUmc) and the London Regional Cancer Program (LRCP) for patients treated with whole brain radiotherapy (20 Gy/5 VUmc, 30 Gy/10 LRCP) with simultaneous integrated boost (SIB) to individual intracranial lesions (40 Gy/5 VUmc, 35-60 Gy/10 LRCP) to perform survival/intracranial control outcome analyses. RESULTS: A total of 120 patients were treated by both the LRCP (n=70) and VUmc (n=50) between 2005 and 2010. Median lesional dose BED3,10 for the entire cohort of patients were 147 and 72 Gy, respectively. Median follow-up for the entire cohort of patients was 4.7 months with median follow-up of 5.2 months for living patients. On multivariable analysis, primary lung cancer (HR 2.044), presence of systemic metastatic disease (HR 1.937), and lower baseline WHO performance status (HR 1.742) were significant (p<0.05) predictors of shorter overall survival. Cumulative brain metastases volume (HR 1.014, p=0.06) was of borderline significance on analysis of intracranial control. CONCLUSIONS: This pooled analysis has provided robust outcome data regarding the use of arc-based radiotherapy with SIB. PMID- 21641068 TI - The dominant negative mutant Artemis enhances tumor cell radiosensitivity. AB - BACKGROUND: Tumor radioresistance often leads to treatment failure during radiotherapy. New strategies like developing radiosensitizer are clinically important. Intervention with DNA double-strand break repair is an effective way to modulate tumor cell radiosensitivity. This study focused on the mutant Artemis fragment-enhanced radiosensitivity of human cervical cancer cells. MATERIAL AND METHODS: We constructed two pEGFP-C1-based eukaryotic expression vectors encoding full-length and the mutant Artemis fragment (D37N-413aa), respectively. HeLa cells were stably transfected with these plasmids or vector. Cell survival was measured by the clonogenic assay. The gammaH2AX foci assay was used to monitor DNA repair after irradiation. Co-immunoprecipitation and Western blot analysis were performed to study protein interaction and phosphorylation of Artemis. RESULTS: Expression of the mutant Artemis fragment (D37N-413aa) delayed DNA DSB rejoining after irradiation, thereby enhanced radiosensitivity of HeLa cell. Further experiments indicate that this mutant Artemis fragment bind to DNA-PKcs and ATM, inhibited phosphorylation of endogenous Artemis, the key molecule for DNA repair and cell radiosensitivity. CONCLUSIONS: The dominant negative mutant Artemis fragment (D37N-413aa) enhanced tumor cell radiosensitivity through blocking activity of endogenous Artemis and DNA repair. It is the first time to modulate tumor cell radiosensitivity via targeting Artemis. This novel mechanism of radiosensitivity strongly suggests the potential role of Artemis in cancer therapy. PMID- 21641069 TI - Expression of thioredoxin system and related peroxiredoxin proteins is associated with clinical outcome in radiotherapy treated early stage breast cancer. AB - BACKGROUND AND PURPOSE: Deregulated redox systems provide cancer cells protection from increased oxidative stress, such as that induced by ionizing radiation. Expression of the thioredoxin system proteins (thioredoxin, thioredoxin reductase and thioredoxin interacting protein) and downstream peroxiredoxins (I-VI), was examined in tumor specimens from early stage breast cancer patients, subsequently treated by breast conserving surgery and locoregional radiotherapy, to determine if redox protein expression is associated with clinical outcome. MATERIAL AND METHODS: Nuclear and cytoplasmic expression was assessed using conventional immunohistochemistry on a tissue microarray of 224 tumors. RESULTS: High expression of cytoplasmic peroxiredoxin-I correlated with a greater risk of local recurrence (p=0.009). When nuclear and cytoplasmic expression patterns were combined, patients with low nuclear but high cytoplasmic expression of peroxiredoxin-I increased significance (p=0.005). Both were independent factors (p=0.006 and 0.003) from multivariate analysis. Associations were obtained between tumor grade and nuclear thioredoxin interacting protein (p=0.01) and with cytoplasmic expression of peroxiredoxin-V (p=0.007) but not with peroxiredoxin-I suggesting that the latter may exert influence via regulation of oxidative stress rather than via altering the tumor phenotype. CONCLUSIONS: Results highlight the potential of using redox protein expression, namely peroxiredoxin-I, to predict clinical outcome and support further studies to validate its usefulness as an independent prognostic, and potentially predictive, marker. PMID- 21641070 TI - Changes in the fraction of total hypoxia and hypoxia subtypes in human squamous cell carcinomas upon fractionated irradiation: evaluation using pattern recognition in microcirculatory supply units. AB - BACKGROUND AND PURPOSE: Evaluate changes in total hypoxia and hypoxia subtypes in vital tumor tissue of human head and neck squamous cell carcinomas (hHNSCC) upon fractionated irradiation. MATERIALS AND METHODS: Xenograft tumors were generated from 5 hHNSCC cell lines (UT-SCC-15, FaDu, SAS, UT-SCC-5 and UT-SCC-14). Hypoxia subtypes were quantified in cryosections based on (immuno-)fluorescent marker distribution patterns of Hoechst 33342 (perfusion), pimonidazole (hypoxia) and CD31 (endothelium) in microcirculatory supply units (MCSUs). Tumors were irradiated with 5 or 10 fractions of 2 Gy, 5*/week. RESULTS: Upon irradiation with 10 fractions, the overall fraction of hypoxic MCSUs decreased in UT-SCC-15, FaDu and SAS, remained the same in UT-SCC-5 and increased in UT-SCC-14. Decreases were observed in the proportion of chronically hypoxic MCSUs in UT-SCC-15, in the fraction of acutely hypoxic MCSUs in UT-SCC-15 and SAS, and in the percentage of hypoxemically hypoxic MCSUs in SAS tumors. After irradiation with 5 fractions, there were no significant changes in hypoxia subtypes. Changes in the overall fraction of hypoxic MCSUs were comparable to corresponding alterations in the proportions of acutely hypoxic MCSUs. There was no correlation between radiation resistance (TCD(50)) and any of the investigated hypoxic fractions upon fractionated irradiation. CONCLUSIONS: This study shows that there are large alterations in the fractions of hypoxia subtypes upon irradiation that can differ from changes in the overall fraction of hypoxic MCSUs. PMID- 21641071 TI - Peptide complex containing GLP-1 exhibited long-acting properties in the treatment of type 2 diabetes. AB - The multiple physiological characterizations of glucagon-like peptide-1 (GLP-1) make it a promising drug candidate for the treatment of type 2 diabetes. However, in vivo, the half-life of GLP-1 is short, which is caused by the degradation of dipeptidyl peptidase-IV (DPP-IV) and renal clearance. Thus, the stabilization of GLP-1 is critical for its utility in drug development. Peptides known as GLP-1 protectors are predicted to increase the half-life of GLP-1 in vivo. Protecting peptides are able to form stable complexes by non-covalent interactions with human GLP-1. In this study, the stability of the complex was investigated, and the physiological functions of the GLP-1/peptide 1 complex were compared to those of exenatide and liraglutide in animals. The results indicated that the GLP 1/peptide 1 complex remarkably raised the half-life of GLP-1 in vivo and showed better glucose tolerance and higher HbA(1c) reduction than exenatide and liraglutide in rodents. Based upon these results, it is suggested that the GLP 1/peptide 1 complex might be utilized as a possible potent anti-diabetic drug in the treatment of type 2 diabetes mellitus. PMID- 21641072 TI - Improved refractory wound healing with administration of acidic fibroblast growth factor in diabetic rats. AB - The aim of the present study is to investigate the effect and mechanism of acidic fibroblast growth factor (aFGF) on treating refractory wound of diabetic rats. SD rats were randomly divided into control group, diabetes group, and aFGF group. Ulcer skin tissues of three groups of rats were respectively collected on days 7 and 14 after establishment of ulcer model for biochemical test, pathological section and immunohistochemistry to comprehensively evaluate the treatment effect of aFGF on diabetic ulcer. The results showed that aFGF could significantly increase capillaries and fibroblast amounts of ulcer tissues, enhance the expression of TGF-beta and PCNA proliferation proteins, and thus improved diabetic ulcer tissues. The preliminary mechanism that aFGF helps to promote healing of diabetic ulcer is possibly associated with that aFGF stimulated ulcer skins to secrete TGF-beta and PCNA proteins and promoted proliferation of capillaries and fibroblasts. PMID- 21641073 TI - More family responsibility, more informal care? The effect of motivation on the giving of informal care by people aged over 50 in the Netherlands compared to other European countries. AB - Against the backdrop of ongoing population ageing, informal care occupies an important place on European political agendas. This article discusses informal caregiving by middle aged and older persons in the Netherlands and other European countries, with particular emphasis on the role played by motives. The data are drawn from SHARE. Our results show that in the Netherlands, it is mainly feelings of being needed and obligation that increase the chance of informal care being given. Deriving pleasure from an activity, by contrast, reduces the likelihood. In Southern Europe, where the responsibility for providing care lies with the family, we found that, contrary to expectations, older carers do not more often feel obliged. They less often report that they feel needed or see being socially active as a way of contributing to society. Our simulations suggest that if the socially active Dutch had the same motives as their Southern European counterparts and behaved similarly in terms of informal caregiving, the number of informal carers would fall. This implies that a greater policy emphasis on family responsibility could actually bring about a decline in the amount of care given, as opposed to the envisaged increase. PMID- 21641074 TI - A review of Health Technology Assessment (HTA) recommendations for drug therapies issued between 2007 and 2009 and their impact on policymaking processes in Poland. AB - OBJECTIVE: The primary objective of this study was to critically review and analyze the Polish Health Technology Assessment (AHTAPol) agency's health technology drug recommendations (HTA activity), in order to ascertain to what extent HTA findings have been incorporated into national drug reimbursement decisions (HTA impact). METHODOLOGY: HTA recommendations issued between 2007 and 2009 were studied. Positive recommendations were classified into three categories: recommendations with major restrictions; minor restrictions; and without restrictions. Definitions of clinical and non-clinical reasons were drawn ups for negative recommendations. The study examined how many different drug technologies assessed by AHTAPol were included in reimbursement lists. RESULTS: In terms of HTA activity, 63 negative and 83 positive HTA recommendations were issued. While clinical arguments were the most prevalent reason for negative HTA recommendations, major restrictions were most common in the positive guidance group. In terms of HTA impact, the results revealed 30 drugs with positive HTA recommendations and four with negative HTA recommendations were included on the reimbursement lists. CONCLUSIONS: Most of AHTAPol's recommendations have a positive outcome for the drug being appraised. The study revealed room for further enhancement of HTA impact. Three key areas that need future attention were identified: consistency, credibility; and pragmatism. PMID- 21641075 TI - Public involvement in the systematic review process in health and social care: a narrative review of case examples. AB - OBJECTIVES: To review the evidence on public involvement in the systematic review process in health and social care; to examine the different methods, levels and stages of involving the public; to synthesise the contributions of the public, as well as the identified tensions, facilitating strategies and recommendations for good practice. METHOD: Systematic literature search and narrative review. FINDINGS: Seven case examples were found covering the following review topics: patients' perspectives on electro-convulsive therapy; user involvement in nursing, midwifery and health visiting research; treatments for degenerative ataxias; teaching, learning and assessment of law in social work education; HIV health promotion for men who have sex with men; the conceptualisation, measurement, impact and outcomes of public involvement in health research; methods of consumer involvement in developing healthcare policy and research, clinical practice guidelines and patient information material. The public was found to contribute to systematic reviews by: refining the scope of the review; suggesting and locating relevant literature; appraising the literature; interpreting the review findings; writing up the review. Numerous tensions, facilitating strategies and recommendations were identified. CONCLUSIONS: The issues raised in this paper should assist researchers in developing and conducting systematic reviews with the involvement of the public. PMID- 21641076 TI - Expression of the IL-23/Th17 pathway in lesions of hidradenitis suppurativa. AB - BACKGROUND: Hidradenitis suppurativa is a debilitating chronic disease primarily affecting intertriginous skin of the axillae, perineum, and inframammary regions. The pathogenesis of this inflammatory disease is still poorly understood. Recently, increased attention has been paid to the role of the immune system. OBJECTIVE: Since the interleukin 12 (IL-12)/T helper 1 (Th1) and the IL-23/Th17 pathways are believed to be crucially involved in the pathogenesis of multiple chronic inflammatory diseases, we investigated the expression and cellular source of IL-12, IL-23, and IL-17 in hidradenitis suppurativa. METHODS: Ten patients with hidradenitis suppurativa were included in the study. Tissue samples were obtained from lesional skin and compared with healthy skin as a control. Expression of IL-12, IL-23, and IL-17 was analyzed by semiquantitative real-time polymerase chain reaction and immunohistochemistry, and the cellular source of these cytokines was determined by double immunofluorescence. RESULTS: IL-12 and IL-23 were found to be abundantly expressed by macrophages infiltrating papillary and reticular dermis of lesional skin. In accordance with the high expression of IL-23 and its important role in the development of T helper 17 (Th17) cells, IL 17-producing T helper cells were found to distinctly infiltrate lesional dermis. LIMITATIONS: The sample size was small. CONCLUSIONS: Our findings suggest that the IL-23/Th17 pathway is expressed in hidradenitis suppurativa and further support involvement of the immune system. Moreover, targeting the IL-12/IL-23 common subunit p40 with novel monoclonal antibodies may represent a new option for the treatment of this recalcitrant disease. PMID- 21641077 TI - Ablative fractional laser therapy as treatment for Becker nevus: a randomized controlled pilot study. AB - BACKGROUND: Becker nevus (BN) is an uncommon pigment disorder characterized by hyperpigmentation and sometimes hypertrichosis. To date, no effective treatment has been available. OBJECTIVES: We sought to assess efficacy and safety of ablative 10,600-nm fractional laser therapy (FLT) in the treatment of BN. METHODS: Eleven patients with BN, older than 18 years, were included in a prospective randomized controlled, observer-blinded split-lesion trial. In each patient two similar square test regions were randomized to either ablative FLT at 10 mJ/microbeam, coverage 35% to 45%, and topical bleaching (to prevent laser induced postinflammatory hyperpigmentation), or topical bleaching alone (to allow comparison of the regions). At 3- and 6-month follow-up, clearance of hyperpigmentation was assessed by physician global assessment, reflectance spectroscopy, melanin index, patient global assessment, patient satisfaction, and histology. RESULTS: At 6-month follow-up, physician global assessment improved in the FLT region (P < .05). Reflectance spectroscopy, melanin index, number of melanocytes, and amount of dermal melanin did not significantly differ between the regions. Patient global assessment and patient satisfaction were 5.0 and 5.9 (visual analog scale score, 0-10), respectively. Side effects were postinflammatory hyperpigmentation (n = 3), erythema (n = 3), burning sensation (n = 3), crusting (n = 3), edema (n = 2), and blistering (n = 2). LIMITATIONS: Limitations include the small number of patients, treatment in spring, possibly suboptimal laser settings, and the combined usage of FLT and a bleaching agent. CONCLUSION: Ablative FLT was moderately effective in some patients with BN. However, postinflammatory hyperpigmentation and relatively negative patient reported outcomes still preclude ablative FLT from being a standard therapy. Larger studies with different laser settings will be required to optimize this treatment modality. PMID- 21641078 TI - Efficacy of mycophenolate mofetil in antimalarial-resistant cutaneous lupus erythematosus. AB - BACKGROUND: Mycophenolate mofetil (MMF) is an immunomodulatory drug shown to be effective in the treatment of systemic lupus erythematosus. Several anecdotal reports have suggested that MMF may be efficacious in the treatment of cutaneous lupus erythematosus (CLE). OBJECTIVES: Our objective was to summarize and report our experience with the use of MMF in patients with CLE recalcitrant to antimalarial therapy. METHODS: We retrospectively analyzed our open-label observations of 24 patients with CLE refractory to antimalarial therapy. The records of all patients visiting the Rheumatic Skin Disease Clinic at the University of Texas Southwestern Medical Center at Dallas from January 1, 2001, to July 1, 2006, were reviewed. RESULTS: MMF was tolerated well and, in conjunction with other therapies, was highly effective in the treatment of antimalarial-resistant CLE. With the addition of MMF to the existing regimen, a majority of patients achieved full control of disease signs and symptoms. All patients experienced some degree of improvement. LIMITATIONS: This is an open label retrospective review. Severity of disease was assessed by qualitative assessment of the clinician. MMF was not used as monotherapy. CONCLUSIONS: Our results indicate that MMF, used as an additional agent in conjunction with standard therapy, is both well tolerated and efficacious in the treatment of refractory CLE. Despite the obvious limitations of the study, we believe this represents further evidence that MMF should be considered early in the treatment of patients refractory to antimalarial therapy. PMID- 21641079 TI - Benchmarking US Department of Veterans Affairs dermatologic services: results from a national survey. AB - BACKGROUND: How well Department of Veterans Affairs (VA) dermatology services provide clinical care, medical education, and innovative research is a largely unexplored topic in the literature. OBJECTIVE: We sought to benchmark VA dermatology services by surveying VA dermatologists about their environment, resources, and the pros and cons of working in the VA. METHODS: Printed surveys were mailed to VA dermatologists and responses were compiled and analyzed. RESULTS: Of 105 dermatology services surveyed, 48% returned surveys completed by board-certified dermatologists (n = 50); 20 surveys completed by nondermatologists were excluded from the analysis. Most services trained dermatology residents (72%) and medical students (80%). One third of services reported significant research involvement. Qualitative analysis revealed the academic environment, patient population, and decreased business management responsibilities as the 3 most commonly cited advantages to VA employment. The most commonly listed disadvantages included low salaries, bureaucracy, and lack of resources. LIMITATIONS: The survey data were self-reported and not independently verified. Not all services returned the survey. CONCLUSIONS: Outpatient VA dermatology services accomplish significant primary care and preventive services (eg, sun safety counseling, skin cancer screening, and treatment). However, the small number of dedicated dermatology services, their irregular geographic distribution, and the lack of staffing and resources may adversely affect optimal patient care. Dermatologist responses regarding the positive and negative aspects of working in the VA system may lead to improved management strategies to better retain and recruit dermatologists to provide patient care, medical education, and medical research despite dramatically lower dermatologist salaries within the VA system compared with private practice. PMID- 21641080 TI - Rituximab for treatment-refractory pemphigus and pemphigoid: a case series of 17 patients. AB - BACKGROUND: Rituximab has been increasingly used in autoimmune blistering dermatoses, mainly in pemphigus. However, only a few larger case series are available on this subject and information on the efficacy of retreatment with rituximab during relapses is lacking. OBJECTIVE: We sought to determine efficacy and adverse effects of adjuvant rituximab. METHODS: Seventeen patients with refractory autoimmune blistering dermatoses (pemphigus vulgaris [PV], n = 8; pemphigus foliaceus [PF], n = 2; bullous pemphigoid [BP], n = 2; mucous membrane pemphigoid, n = 5) were treated 4 times with rituximab at an individual dose of 375 mg/m(2) in weekly intervals or twice with 1000 mg 2 weeks apart. Six of 8 patients with a relapse after this regimen received rituximab again twice with 1000 mg in a 2-week interval. RESULTS: All lesions cleared in 14 patients (7 PV, two PF, two BP, 3 mucous membrane pemphigoid), whereas partial healing was found in 3 others (one PV, two mucous membrane pemphigoid). Relapses occurred in 8 patients (5 PV, two PF, one BP). Retreatment with rituximab again resulted in complete (two PV, one PF, one BP) or partial (two PV) remission. Serious side effects associated with rituximab were not observed. LIMITATIONS: Rituximab has been combined with various other immunosuppressive or immunomodulatory treatments. CONCLUSION: Adjuvant rituximab is effective and well tolerated not only in patients with pemphigus but also with pemphigoid. Efficacy and safety of rituximab are maintained when it is readministered during relapses. PMID- 21641081 TI - Radiotherapy alone for Merkel cell carcinoma: a comparative and retrospective study of 25 patients. AB - BACKGROUND: Merkel cell carcinoma (MCC) is a rare skin cancer. Cumulative data from retrospective series support the notion that benefits are obtained by both wide excision and adjuvant radiation therapy. However, surgery may be difficult to perform with tumors located in the head and neck region and/or in elderly patients with comorbidities incompatible with general anesthesia. OBJECTIVE: We assessed the benefit of treating MCC exclusively with radiation when conventional treatment (surgery followed by radiotherapy) is not possible. METHODS: A total of 25 patients with primary MCC were treated at our institution exclusively with radiotherapy. Because there is no consensus about this specific approach, we compared the recurrence rate of the 25 patients receiving radiotherapy alone with that of 25 patients who received conventional treatment at our institution. RESULTS: The median follow-up periods were 3 years (range: 5 months-11 years) for the group receiving only radiotherapy (group 1) and 9 years (range: 12 months-16 years) for the conventional therapy group (group 2). No local relapses were observed, but two locoregional relapses were observed in group 1, and 4 in group 2. No statistical differences were found in overall and disease-free survival between the two groups of patients. LIMITATIONS: The limitation of this study is its retrospective nature. CONCLUSIONS: This study confirms the results of our previous research demonstrating that it is possible to treat inoperable MCC exclusively with radiotherapy to obtain an outcome similar to that which is achievable with conventional treatment. PMID- 21641082 TI - Studying the transmission dynamics of meticillin-resistant Staphylococcus aureus in Hong Kong using spa typing. AB - This study investigated the transmission dynamics of meticillin-resistant Staphylococcus aureus (MRSA) in a tertiary referral surgical unit with 300 beds. All adult patients were actively screened for MRSA by culture at hospital admission and twice weekly thereafter during hospitalisation from 1 October to 31 December 2008. The colonisation pressure per 1000 patient-days and the incidence density of nosocomial MRSA transmission per 1000 colonisation-days were calculated for the different spa types of MRSA. In total, 6619 nasal swabs were obtained from 2289 patients. One-hundred and forty-eight (7%) patients had MRSA in nasal swabs at admission screening, of which 68/148 (46%) were residents of elderly care homes. Fifty-two of 2141 (2%) patients had conversion of nasal MRSA carriage status from negative to positive during hospitalisation. Among the 200 patients with MRSA, spa types t1081 and t037 were found in 99 (50%) and 30 (15%) patients, respectively. The colonisation pressure per 1000 patient-days was 40.9 for t0181, 22.2 for t037 and 26.3 for the less common spa types. The incidence densities of nosocomial MRSA transmission per 1000 colonisation-days were significantly higher for t1081 (28.5 vs 4.0, P<0.01) and t037 (21.5 vs 4.0, P=0.03) compared with the less common spa types. Proactive screening of MRSA in patients from elderly care homes and targeted isolation of these patients, especially those carrying spa types with high transmissibility, are important for the control of MRSA in hospitals. PMID- 21641083 TI - Klebsiella pneumoniae producing KPC carbapenemase in a district general hospital in the UK. AB - We report two patients with multidrug-resistant KPC-carbapenemase-producing Klebsiella pneumoniae urinary tract infections. A bla(KPC-2) gene was detected in both of the isolates by polymerase chain reaction and sequencing. The isolates had identical pulsed-field gel electrophoresis patterns and belonged to sequence type ST11. The index patient probably acquired the KPC-producing strain while in hospital in Curacao, with subsequent nosocomial transmission to the second patient occurring in our hospital. We describe the interventions that were taken to prevent its further spread within the acute Trust and the community. PMID- 21641084 TI - Study on the effectiveness of disinfection with wipes against methicillin resistant Staphylococcus aureus and implications for hospital hygiene. AB - BACKGROUND: Environmental contamination with methicillin-resistant Staphylococcus aureus (MRSA) poses a significant risk in the epidemiology of infection. This study aimed to assess the effectiveness of disinfection in clinical areas by assessment of the wipe-rinse method to MRSA in the immediate patient environment, on both the bed rails and the cleaning wipes. METHODS: The environmental disinfection procedures of 8 MRSA-positive hospitalized patients in 2 orthopedic wards were evaluated. A total of 56 pre- and postdisinfection samples from the bed rails as well as pre- and postuse wipes samples were collected. RESULTS: The average MRSA loads on the bed rails before and after disinfections were 4.4 colony-forming units (cfu)/cm(2) and 0.4 cfu/cm(2), respectively, the process reducing MRSA survival from a mean of -0.70 log to -1.65 log. The mean difference of MRSA survival detected on the wipes after each wiping ranged from 0.13 to 0.42 log. CONCLUSION: The presence of MRSA in the proximity of the patient, ie, the bed rails as well as the cleaning tool (the wipe), was demonstrated in this study. If thorough rinsing was not conducted between wiping, bacteria accumulated on the wipes, which can result in cross transmission. PMID- 21641085 TI - Mumps exposure of a health care provider working in a neonatal intensive care unit leads to a hospital-wide effort that prevented an outbreak. AB - BACKGROUND: Control measures were instituted in a neonatal intensive care unit (NICU) in Syracuse, New York, when a neonatologist became ill with mumps after returning from Africa. Two health care providers (HCPs) who worked with the neonatologist developed parotitis within 13 days of exposure. Outbreak control included furloughing the neonatologist and the 2 HCPs until after 5 days of the onset of parotitis, cohorting and isolating all exposed infants in the NICU, and implementing droplet precautions. All susceptible HCPs were immunized, and all HCPs were required to wear surgical masks when within 3 feet of patients. RESULTS: Five HCPs developed parotitis. The neonatologist and 2 of the HCPs were confirmed cases, and 2 other HCPs did not meet the case definition. Twenty-six HCPs who worked in other units of the hospital besides the NICU developed nonspecific signs and symptoms of illness. Of the 2,904 HCPs tested, 287 (10%) had negative antibody results, and 153 (8%) were age 40 years or older. Of the 287 HCPs with negative antibody titers, 200 (70%) received the mumps-measles rubella vaccine in response to this effort. No cases of mumps were reported in exposed infants, children, or adult patients during the time of exposure. CONCLUSION: Infection control efforts, including vaccinating susceptible HCPs and instituting droplet precautions, might have prevented mumps infection in the NICU patients. PMID- 21641086 TI - The long-term outcome of a multifaceted intervention to reduce ventilator associated pneumonia: can zero really be achieved? PMID- 21641087 TI - Bacterial contamination of glucose test strips: not to be neglected. PMID- 21641088 TI - Reduction of catheter-related bloodstream infections through the use of a central venous line bundle: epidemiologic and economic consequences. AB - BACKGROUND: Central venous lines (CVLs) are used extensively in intensive care units (ICUs) but can sometimes lead to catheter-related blood stream infections (CRBSIs). This study evaluated a "CVL bundle" to see whether the CRBSI rate would decrease, analyze any changes in the flora of CRBSIs, and project any decrease in health care costs. METHODS: The CVL bundle was implemented on all patients admitted to the ICU starting January 2008. Data from CRBSI rates from 2006 and 2007 were pooled to compare the intervention. A Poisson analysis generated a relative risk reduction. Determination of costs were made by taking the excess length of stay multiplied by other costs (supplies, medications, cost of replacement of CVL) at our institution. RESULTS: Overall infection rates decreased with an improvement in CRBSIs in all ICUs that participated. Although the proportion of gram-negative organisms did not change significantly, there was a decrease in the proportion of gram-positive infections (P = .05) and an increase in fungal infections (P = .04). The total excess cost per organism was determined by the following: total excess cost = excess length of stay + replacement of CVL + drug administration + antibiotic cost. The weighted excess cost took the total excess cost times a correction factor based on organism frequency. The total excess cost of any given CRBSI is approximately $32,254. CONCLUSION: Preventing CRBSIs can improve patient care while reducing hospital stays, costs, and possible mortality. CVL bundles are fairly easy to perform with reproducible results. PMID- 21641089 TI - Reduction in the microbial load on high-touch surfaces in hospital rooms by treatment with a portable saturated steam vapor disinfection system. AB - BACKGROUND: Recent scientific literature suggests that portable steam vapor systems are capable of rapid, chemical-free surface disinfection in controlled laboratory studies. This study evaluated the efficacy of a portable steam vapor system in a hospital setting. METHODS: The study was carried out in 8 occupied rooms of a long-term care wing of a hospital. Six surfaces per room were swabbed before and after steam treatment and analyzed for heterotrophic plate count (HPC), total coliforms, methicillin-intermediate and -resistant Staphylococcus aureus (MISA and MRSA), and Clostridium difficile. RESULTS: The steam vapor device consistently reduced total microbial and pathogen loads on hospital surfaces, to below detection in most instances. Treatment reduced the presence of total coliforms on surfaces from 83% (40/48) to 13% (6/48). Treatment reduced presumptive MISA (12/48) and MRSA (3/48) to below detection after cleaning, except for 1 posttreatment isolation of MISA (1/48). A single C difficile colony was isolated from a door push panel before treatment, but no C difficile was detected after treatment. CONCLUSION: The steam vapor system reduced bacterial levels by >90% and reduced pathogen levels on most surfaces to below the detection limit. The steam vapor system provides a means to reduce levels of microorganisms on hospital surfaces without the drawbacks associated with chemicals, and may decrease the risk of cross-contamination. PMID- 21641090 TI - [Laparoscopic partial nephrectomy]. AB - OBJECTIVES: To analyze the surgical and oncologic outcome of prospective experience with laparoscopic partial nephrectomy. We describe the surgical technique and mid term oncological results achieved. MATERIAL AND METHODS: 60 patients were operated with this technique between June 2005 and June 2009. The mean age of patients was 58.9 [38-77] years, being 40 (66.7%) males and 20 (33.3%) women. The average BMI was 26.8 [18-40]. Laterality was 28 (46.7%) tumors rights and 32 (53.3%) left, being located in the upper pole in 14 (23.3%) patients, in the middle third in 13 (21.7%) in the lower pole in 22 (36.7%) and hiliar region in 11 (18.3%). In 23 cases (38.3%) tumors were located in the anterior valve, in 24 (40%) in posterior valve, in 10 (16.7%) at the outer edge and 3 (5%) at the inner edge. The average size tumor on CT was 3.3 [1-6.4] cm and in the surgical specimen 3.1 [1.2-7] cm. RESULTS: The mean operative time was 107.17 [50-185] min, with a warm ischemia time of 33 [0-70] min. In 56 cases (93.3%) had a single artery and 4 (6.7%) cases had 2 arteries. The artery was clamped alone in 15 patients (25%), artery and vein in 44 (73.3%) and no clamping was performed in 1 (1.7%). We repaired the urinary tract in 32 patients (53.3%), leaving ureteral catheter in all patients. 20% of patients (12) required transfusion. Intraoperative complications occurred in 5 patients (8.7%). These were: 1 splenic injury requiring splenectomy (1.7%), 1 tear in the vena cava, sutured laparoscopically (1.7%) and 3 cases of bleeding due to bulldog malfunction (5%). Postoperative complications occurred in 11 patients (18.7%) and these were: 1 wall hematoma that required reoperation (1.7%), 1 urinary fistula ending in renal atrophy and subsequent nephrectomy (1.7%), 3 intracavitary hematomas hich resolved conservatively (5%), 1 arteriovenous fistula that needed embolization (1.7%), 1 urinoma that was resolved with percutaneous drainage (1.7%) and 3 cases of postoperative fever (5%). Margins were positive in 1 patient (1.7%). In 49 cases (81.7%) histology was renal cell carcinoma, in 8 (13.3%) oncocytoma, in 2 (3.3%) angiomyolipoma and 1 (1.7%) metastasis. The average stay was 5 [3-29] days. Median follow up was 31 [12-61] months. There was a local recurrence at 16 months (hiliar primary tumor 2.5 cm) and an ipsilateral adrenal metastasis at 34 months (primary tumor 5.6 cm in left lower pole). CONCLUSIONS: In this series of laparoscopic partial nephrectomy low rate of complications, good oncologic results and low recurrence rate in the short term are shown. More patients and further monitoring is required to strengthen the functional and oncological outcomes of this surgical technique. PMID- 21641091 TI - Cushing's syndrome due to ectopic ACTH. Contribution of the somatostatin receptor scintigraphy. PMID- 21641092 TI - [Usefulness of 18F-FDG PET-CT in the presurgical assessment of malignant pleural mesothelioma treated with neoadjuvant chemotherapy]. AB - Malignant pleural mesothelioma is a relatively rare, but highly aggressive, tumor, associated to exposure to asbestos, with a life expectancy between 9 and 17 months. Chest pain and dyspnea are the most frequent symptoms. The most commonly used therapy is surgery accompanied by chemotherapy. Preoperative assessment, after chemotherapy, has been done using magnetic resonance imaging and computed tomography (CT). However, these techniques cannot predict early response to therapy, because of the slow structural change of the tumor. The aim of this case report is to review and learn about the growing use of PET-CT imaging with (18)F-FDG in the preoperative staging of malignant pleural mesothelioma and its influence in selecting the most appropriate type of surgery. PMID- 21641093 TI - 7-Substituted pterins provide a new direction for ricin A chain inhibitors. AB - Ricin is a potent toxin found in castor seeds. The A chain, RTA, enzymaticlly depurinates a specific adenosine in ribosomal RNA, inhibiting protein synthesis. Ricin is a known chemical weapons threat having no effective antidote. This makes the discovery of new inhibitors of great importance. We have previously used 6 substituted pterins, such as pteroic acid, as an inhibitor platform with moderate success. We now report the success of 7-carboxy pterin (7CP) as an RTA inhibitor; its binding has been monitored using both kinetic and temperature shift assays and by X-ray crystallography. We also discuss the synthesis of various derivatives of 7CP, and their binding affinity and inhibitory effects, as part of a program to make effective RTA inhibitors. PMID- 21641094 TI - [Hypertrophic osteoarthropathy and aortic graft infection: Two cases]. AB - INTRODUCTION: Hypertrophic osteoarthropathy (HOA) located to lower extremities may be the initial manifestation of an infected aortic graft. CASE REPORTS: We report two patients with HOA secondary to aortobifemoral vascular prosthesis infection and aortoenteric fistula. As reported in the literature, initial manifestations included fever, painful swelling of limbs, joint pain, clubbing and in one case intestinal bleeding. These symptoms preceded frank episodes of septicaemia due to a wide variety of bacteria (microbial enteric flora). Bone scan was an appropriate tool for confirming the diagnosis of HOA. Abdominal computed tomography, and PET-scan were useful for detecting vascular infection. Aortoenteric fistula remained difficult to identify. CONCLUSIONS: Mechanisms involved in the pathogenesis of HOA associated with infected graft are poorly understood, but vascular endothelial growth factor (VEGF) could play a major role. The mortality of infected aortic grafts remains high, but knowledge of this association could lead to early diagnosis and accurate treatment. PMID- 21641095 TI - Continuing professional development programmes for rural nurses involved in palliative care delivery: an integrative review. AB - PURPOSE: To review published studies evaluating the impact of continuing professional development (CPD) programmes on rural nurses palliative care capabilities in order to inform the development of targeted learning activities for this population. DESIGN: An integrative review. METHODS: Searches of key electronic databases and the World Wide Web was undertaken using key words, followed by hand searching for relevant articles. All studies were reviewed by two authors using a critical appraisal tool and level of evidence hierarchy. RESULTS: The search strategies generated 74 articles, with 10 studies meeting the inclusion criteria. All of these studies evaluated palliative care CPD programmes involving rural nurses which focused on increasing palliative care capabilities. The evidence generated by this review was limited by the absence of randomised controlled trials. A level III-1 study, with a small sample size provided the highest level of evidence, but the lack of control negated the investigators' capacity to confirm causality. Few studies measured the impact of CPD on the quality of care or utilised novel technology to address the tyranny of distance. Despite, these limitations valuable insights into the barriers and facilitators to engaging rural nurses in palliative care learning opportunities were identified. CONCLUSIONS: Evidence that CPD impacts positively on patient and families outcomes is necessary to sustain an on-going investment in learning activities. In order to optimise the opportunities afforded by emerging web-based technology rural nurses' need to develop and maintain their computer competencies. Further investigation of the impact of specialist clinical placements on rural nurses' palliative care capabilities is also indicated. PMID- 21641096 TI - Social capital and the nursing student experience. AB - The article develops a theoretical argument relating to the first year experience of nursing students based on the findings from a PhD study. A 'Connections Continuum' is a tool that was developed from its qualitative and quantitative findings. The tool aims to describe the aspects of a nursing student's first year experience that are of consequence and which impact on student success. The overall findings of the wider PhD study reveal that, usually, expectations and experiences should correspond. In addition, the thematic findings expose the importance of connections - with people, with the classroom, with practice, and with professional education. These key aspects are described within the continuum with the assertion that, rather obviously, the student is more likely to be successful (in terms of staying on the course and engaging with the overall experience) when placed towards the positive end of the continuum. A theoretical argument is put forward that draws on social capital as a theoretical 'backbone' for the Connections Continuum and an argument is developed that points to social capital (within the practical framework of the Connections Continuum) as an approach that contributes to enhancement of the nursing student experience at all levels. PMID- 21641097 TI - Vitamin D status in children and young adults with perinatally acquired HIV infection. AB - BACKGROUND & AIMS: To assess vitamin D (Vit-D) [serum 25-hydroxyvitamin D (25 OHD) concentrations] in children and young adults with perinatally acquired HIV compared to geographically similar healthy children. METHODS: 25-OHD in children and young adults with HIV was compared to a healthy group. Vit-D deficiency and insufficiency were defined as 25-OHD <11 ng/mL and 25-OHD <30 ng/mL, respectively. RESULTS: Children with HIV (n = 81), mean age 13.8 +/- 4.1 years, 48% female, 83% Black, were compared to healthy subjects (n = 372), mean age 12.4 +/- 3.4 years, 51% female, 37% Black. For the HIV group, 84% were on HAART, 54% had plasma HIV RNA <400 cpm, and 35% had moderate to severe immunosuppression (CD4+ count <500 cells/mm). Vit-D deficiency/insufficiency was present in 36% and 89% of those with HIV, and 15% and 84% of the comparison group, respectively. Vit D deficiency was more prevalent in those with HIV (unadjusted odds ratio: 3.25; 95% CI: 1.9-5.5). For both groups, prevalence of Vit-D deficiency increased with age, BMI Z-score, Black race, and in winter/spring months. Vit-D deficiency was associated with a greater degree of immunosuppression in the subjects with HIV. CONCLUSIONS: Vit-D deficiency was increased in subjects with perinatally acquired HIV and may be associated with disease severity. PMID- 21641098 TI - Twelve steps per foot are recommended for valid and reliable in-shoe plantar pressure data in neuropathic diabetic patients wearing custom made footwear. AB - BACKGROUND: Dynamic in-shoe plantar pressure assessment is used both in research and clinical practice to evaluate therapeutic footwear interventions in neuropathic diabetic patients. The aim was to determine the required number of footsteps for reliable and valid in-shoe plantar pressure data in these patients. METHODS: In 30 neuropathic diabetic patients wearing custom-made therapeutic footwear, in-shoe plantar pressures were measured for a minimum of 20 midgait walking steps per foot. For each incremental number of steps and for each of six anatomical regions per foot, peak pressure, pressure-time integral, contact area, contact time, and force-time integral were calculated. Reliability was assessed by calculating intraclass correlation coefficients. Validity was assessed by calculating the coefficient of variation between each n-step protocol and the 20 step reference protocol based on Limits of Agreement analysis. Data was considered reliable with intraclass correlation coefficients >0.90 and valid with coefficients of variation <10%. FINDINGS: Three steps per foot were required to obtain reliable data for each foot region and parameter. Depending on the parameter, between 7 and 17 steps per foot were required to obtain valid data. With the exception of deviant outcomes in three forefoot regions for force-time integral, overall 12 steps per foot were required for valid data. INTERPRETATION: For neuropathic diabetic patients wearing custom-made therapeutic footwear, 12 midgait steps per foot are required to obtain valid and reliable in-shoe plantar pressure data. This provides directions for the use of in-shoe plantar pressure analysis in research and clinical practice in this patient group. PMID- 21641099 TI - Stem cell transplantation for indolent lymphoma: a reappraisal. AB - Allogeneic transplantation is established as a curative treatment for follicular lymphoma, but with considerable short and long-term morbidity and mortality. Data and controversies regarding conditioning regimen, donor source, GVHD prophylaxis, post transplant interventions and approaches to predict and reduce morbidity and mortality are reviewed. Total body irradiation is very effective but toxic and reduced intensity conditioning is often preferred though associated with somewhat higher rates of recurrence. The risk of chronic GVHD and its late sequelae can be markedly reduced by in-vivo T-cell depletion using alemtuzumab but also leads to somewhat higher incidence of disease recurrence. When using such treatment strategies, one can consider prophylactic or preemptive donor lymphocyte infusions or low toxicity medical treatment such as rituximab. Overall the long term outcomes, particularly survival and current progression free survival of patients undergoing allogeneic transplantation for indolent lymphoma have steadily improved and transplant can now often safely be considered up to the sixth decade of life. Outcomes of unrelated donor transplantation approach those of HLA-identical sibling transplant and even mismatched umbilical cord transplant can be considered in selected patients. The assessment of risks and benefits is aided by the use of various novel tools. PMID- 21641100 TI - [Changing epidemiology of native valve infective endocarditis]. AB - INTRODUCTION AND OBJECTIVES: The aim of our study is to assess changes in the epidemiologic features of patients with native valve infective endocarditis. METHODS: We analyzed a prospective series of 228 cases of native valve infective endocarditis in non-intravenous drug users attending our center between 1987 and 2009. We compared three subperiods: 1987-1994 (67 cases), 1995-2002 (74 cases) and 2003-2009 (87 cases). RESULTS: The mean age of patients has progressively increased (38+/-22 years in the first subperiod vs 60+/-16 years in the third; P<.001), as has the proportion of cases without predisposing heart disease (25%, 46% and 67%; P<.001). Incidence of mitral valve prolapse remained stable (12%, 18% and 11%). Percentages of patients with predisposing heart disease and who were aware of their condition have fallen in recent years (45%, 27% and 21%; P<.001). A portal of entry for the infection could not be identified in 64%. Overall, Staphylococcus aureus is the most frequent causative organism (26%) whereas the percentage of cases caused by Streptococcus viridans remains unaltered (22%, 20% and 24%). CONCLUSIONS: We found significant changes in the epidemiology of native valve infective endocarditis. The incidence of patients without predisposing heart disease has increased significantly and staphylococci are the most frequent causative organisms. Full English text available from: www.revespcardiol.org. PMID- 21641101 TI - Evidence to support the single-dose over the two-dose protocol for routine antenatal anti-D Rhesus prophylaxis: a prospective observational study. AB - OBJECTIVE: To assess the potential benefit of the single-dose compared with two dose protocol for routine antenatal anti-D prophylaxis (RAADP). STUDY DESIGN: Prospective observational study during February to October 2009 in a district teaching hospital delivering 6000 women annually. Obstetric notes of eligible RhD negative women were reviewed following delivery to identify gestation and dose for RAADP injections, with laboratory confirmation when necessary. The primary outcome was compliance administering RAADP at the appropriate gestation with the secondary analysis of compliance for nulliparae and multiparae. RESULTS: For 644 eligible women, there was no statistically significant difference in administering the 28 week injection between the two-dose (95%) and single-dose (98%) protocols (P=0.17, OR 2.47: 95%CI (0.73, 8.34)). Compliance giving the injection at the correct gestation at 28 +/- 1 weeks was achieved in 78% and 77% respectively (P=0.72, OR 1.10: 95%CI (0.69, 1.74)). For the two-dose protocol the second injection was given to 96% of those who had received the first injection, but at the correct time point in only 67%. By eliminating the variability of the 34 week injection, the single-dose protocol resulted in significantly greater overall success in giving RAADP at the appropriate times compared with the two dose protocol (P=0.021, OR 1.68: 95%CI (1.08, 2.64)). There were no differences between nulliparae and multiparae for both protocols. CONCLUSION: The single-dose protocol provides enhanced compliance over the two-dose protocol with the potential for reduced sensitisation combining economic and manpower benefits. There remains room for improvement in administering RAADP at the correct gestation. PMID- 21641102 TI - Cesarean delivery for first pregnancy and neonatal morbidity and mortality in second pregnancy. AB - OBJECTIVE: To examine impact of cesarean delivery in first pregnancy on neonatal mortality and morbidity in second pregnancy. STUDY DESIGN: Retrospective cohort study using 1995-2002 US birth registration data. Neonatal mortality and morbidity in second pregnancy of cesarean deliveries in the first pregnancy were compared with vaginal deliveries in the first pregnancy. RESULTS: A total of 9,643,175 singleton second births were eligible in the analysis after excluding those with unknown delivery method (1,801,339 with a previous cesarean delivery and 7,841,836 with a previous vaginal delivery). Compared with vaginal delivery group, infants born to mothers with a previous cesarean delivery had increased risks of assisted ventilation (OR=1.47, 95% CI 1.46, 1.49), low Apgar's score (OR=1.14, 95% CI 1.12, 1.17), seizure (OR=1.36, 95% CI 1.27, 1.45), fetal distress (OR=1.46, 95% CI 1.44, 1.47), and asphyxia-related neonatal death (OR=1.40, 95% CI 1.29, 1.52). The association between mode of delivery in first pregnancy and neonatal outcomes in second pregnancy remained the same after excluding women with chronic health problems or adverse birth history and adjusting for potential confounding factors. CONCLUSION(S): Cesarean delivery in first pregnancy is associated with increased risks of neonatal morbidity and mortality in second pregnancy. PMID- 21641103 TI - Maternal angiogenic profile in pregnancies that remain normotensive. AB - OBJECTIVE: We sought to determine if maternal characteristics are associated with angiogenic profile in the first and second trimester of normotensive pregnancies. STUDY DESIGN: Circulating levels of maternal placental like growth factor (PlGF), soluble fms-like tyrosine kinase receptor (sFlt1), and soluble endoglin (sEng) were measured in serum samples collected during the first (median 11.3 weeks) and second trimester (median 17.1 weeks) of 182 normotensive pregnancies. Diastolic blood pressure (DBP), systolic blood pressure (SBP), and mean arterial pressure (MAP) were measured at the same visits when samples were collected to measure angiogenic factors. Linear regression analysis was used to examine associations of the angiogenic measures with maternal characteristics. The association between blood pressure measures and concentrations of angiogenic factors was evaluated using Spearman correlation and linear regression analysis. RESULTS: In adjusted analyses, nulliparous women had higher sFlt1 concentrations in both first (p=0.06) and second (p=0.001) trimester. Higher BMI was associated with greater sFlt1 concentrations in both the first (p=0.004) and second trimester (p=0.008), but significantly lower sEng concentrations in both trimesters (p=0.002 for first trimester and p=0.0009 for second). Nulliparity and higher BMI also were associated with higher sFlt1/PLGF anti-angiogenic ratios in both first (p=0.05 and p=0.007, respectively) and second trimesters (p=0.003 and p=0.02, respectively). First trimester sFlt1 levels were weakly correlated with first trimester SBP (r(s)=0.18, p=0.03) and MAP (r(s)=0.16, p=0.04). Second trimester sEng levels were inversely associated with second trimester MAP (r(s)=-0.17, p=0.05). Including blood pressure measures in the linear regression models did not change the reported associations of angiogenic factors with maternal characteristics. CONCLUSIONS: These results demonstrate that even early in normotensive pregnancies maternal characteristics are associated with variations in angiogenic profile across this population. PMID- 21641104 TI - Prevention of pre-eclampsia with low-dose aspirin or vitamins C and E in women at high or low risk: a systematic review with meta-analysis. AB - The aim of this study is to review literature about the efficacy of low dose aspirin (LDA) and vitamins C/E (VCE) to prevent pre-eclampsia in women at high and low risk. Randomized clinical trials were included and stratified for high and low risk women. Inclusion criteria were: assignment of patients in treated or placebo groups, definition of pre-eclampsia according to the guidelines of the International Society for the Study of Hypertension in Pregnancy. Exclusion criteria were: omitting at least one of the inclusion criteria, trials involving women with pre-eclampsia at trial entry, studies investigating hypertensive disorders other than pre-eclampsia, prophylaxis of intrauterine growth restriction with low-dose aspirin or vitamins C/E, non-randomized studies and data reported in graphs or percentages. The incidence of pre-eclampsia, perinatal outcomes and adverse effects attributable to LDA and VCE were compared between treated women and placebo. Inter-studies heterogeneity was tested. P<0.05 was considered significant. pooled odds ratios (OR) with 95% confidence intervals (95% CI) were calculated. PRISMA guidelines were followed. Fifteen studies were pooled. LDA did not decrease the incidence of pre-eclampsia in high-risk (396/5025 - 8% vs placebo: 464/5027 - 9%; P=0.05; OR: 0.72; 95% CI: 0.51-1.00) and low-risk (137/4939 - 3% vs placebo: 166/4962 - 3%; P=0.10; OR: 0.82; 95% CI: 0.65-1.04) women. Similarly, VCE did not reduce the incidence of pre-eclampsia in high-risk (VCE: 250/1744 - 14% vs placebo: 275/1741 - 16%; P=0.24; OR: 0.84; 95% CI: 0.63-1.12) and low-risk (VCE: 56/935 - 6% vs placebo 47/942 - 5%; P=0.57; OR: 1.20; 95% CI: 0.82-1.75) women. In high-risk women, other hypertensive disorders were more frequent in VCE (121/1692 - 7%) than placebo (79/1693 - 5%; P=0.002). Perinatal outcomes were not improved by LDA or VCE. CONCLUSION: there is no evidence to support the administration of LDA or VCE to prevent pre-eclampsia. PMID- 21641105 TI - Maternal heart rate changes during labour. AB - OBJECTIVES: Labour and delivery represent a considerable effort for pregnant women. Lack of aerobic fitness may limit pushing efforts during childbirth and represents increased cardiovascular strain and risk. Increasing prevalence of sedentary behaviour and lack of aerobic fitness may reduce heart rate reserve during labour. STUDY DESIGN: We quantified maternal heart rate reserve (maximum heart rate minus resting heart rate) of 30 healthy pregnant women during labour and delivery and related it to habitual daily physical activity levels quantified during the third pregnancy trimester by the Pregnancy Physical Activity Questionnaire. RESULTS: Heart rates during labour reached values similar to those observed during moderate to heavy physical exercise. During active pushing one out of five women reached heart rates more than 90% of their heart rate reserve (188 +/- 7 beats per min). Half of the women reached more than 70% of heart rate reserve (172 +/- 14 beats per min). Physically inactive women used more of their heart rate reserve as physically more active women (87 +/- 20% vs. 65 +/- 12%, upper and lower tertile respectively, p<0.05). CONCLUSIONS: Use of heart rate reserve for the effort of labour is increased in physically inactive women and may potentially limit the intensity and duration of pushing efforts. Such higher cardiovascular strain in physically less active women may represent increased cardiovascular risk during labour. PMID- 21641106 TI - Evaluation of a decision protocol for type of delivery of infants in breech presentation at term. AB - OBJECTIVES: To assess the safety of a decision protocol for type of delivery of breech presentations after 37 weeks of gestation. This protocol was based on a comparison of fetal head and maternal pelvic measurements. STUDY DESIGN: Retrospective before-and-after study of 1133 breech infants born between 2000 and 2008 in a tertiary centre, analysing neonatal condition and percentage of vaginal deliveries by study period. RESULTS: Application of the protocol was accompanied by an increase in the percentage of vaginal deliveries of breech presentations from 24.0% in 2000-2004 to 38.5% in 2004-2008 (p<0.001), without impairment of neonatal condition [composite mortality/morbidity variable: 3/567 (0.5%) vs 4/566 (0.7%), respectively; p>0.99; neonatal arterial pH<7.0: 8/521 (1.5%) vs 4/529 (0.8%), respectively; p=0.23]. CONCLUSION: The use of a protocol based on objective criteria at the authors' centre made it possible to increase the percentage of vaginal deliveries of breech presentations without impairing neonatal condition. PMID- 21641107 TI - A prospective comparison of single-port laparoscopically assisted vaginal hysterectomy using transumbilical GelPort access and multiport laparoscopically assisted vaginal hysterectomy. AB - OBJECTIVE: To evaluate the feasibility and safety of single-port laparoscopically assisted vaginal hysterectomy (SP-LAVH) using transumbilical GelPort access. STUDY DESIGN: A prospective case-control study was performed at a University teaching hospital between January 2009 and March 2010, a total of 242 women with a uterus <=16 weeks gestational size were enrolled in the study. Eighty women underwent SP-LAVH using transumbilical GelPort access (SP-LAVH group), and 162 women underwent conventional multiport LAVH (conventional LAVH group). RESULTS: There were no statistical differences between groups in the patients' demographic characteristics, median operating time (92.5 vs. 90 min; P=0.479), postoperative changes in hemoglobin concentration (1.4 vs. 1.4 g/dL; P=0.290), weight of the resected uterus (246 vs. 256 g; P=0.098), return of bowel activity (37.1 vs. 39.8h; P=0.103), hospital stay (3 vs. 3 days; P=0.554), complication rate (3.8 vs. 4.3%; P=1.000), and the rate of using an additional trocar or conversion to laparotomy (1.3 vs. 0.6%; P=0.553). CONCLUSIONS: SP-LAVH using transumbilical GelPort access is feasible and safe in women with a uterus <= 16 weeks gestational size. However, a large prospective randomized study is needed to confirm this conclusion and to establish guidelines for the use of SP-LAVH. PMID- 21641108 TI - Prevalence and incidence of depressive and anxious symptoms in couples undergoing assisted reproductive treatment in an Italian infertility department. AB - OBJECTIVE: We have conducted a longitudinal observational study in order to evaluate the prevalence and the incidence of depressive and anxious symptoms in women and men seeking infertility treatment and to analyze associated factors or risk factors for these kinds of disorders. STUDY DESIGN: A total of 1000 consecutive couples that visited our center for the first time were asked to join this study. Depressive and anxious symptoms were assessed with self-rating Zung Depression Scale (ZDS) and Zung Anxiety Scale (ZAS) questionnaires. A second assessment was planned at the time of beta HCG dosage (or at the moment of cycle suspension). A standard questionnaire was used to investigate socio-demographic information and the psychological aspects of couples undergoing in vitro fertilization (IVF) treatment. The fertility history and outcome of IVF treatment were collected from patients' medical records. RESULTS: 14.7% of women had anxious symptoms and 17.9% depressive symptoms, whereas 4.5% of men had anxious symptoms and 6.9% depressive symptoms. Women with depressive and anxious symptoms were younger, more often had an anxious partner and had a longer history of infertility. Men with depressive and anxious symptoms more frequently had a temporary job, they had an anxious partner and they were more frequently at the first in vitro fertilization (IVF) cycle. The incidence of depressive and/or anxious symptoms was 18.5% in women and 7.4% in men. Age and previous IVF treatments seem not to be associated with incidence of depressive or anxious symptoms. CONCLUSION: Both the prevalence and incidence of depressive and/or anxious symptoms in couples undergoing IVF treatment were worthy of note and should not be underestimated. More attention must be paid to psychological aspects in young women and in couples with a long history of infertility or previous failure treatments. Having an anxious partner was associated with anxious and depressive symptoms. For this reason, both males and females might benefit from psychological intervention, which could also help them to be supportive to each other. PMID- 21641109 TI - Stakeholder participation within the public environmental system in Chile: major gaps between theory and practice. AB - The main objective of this paper is to present a critical analysis of the stakeholder participation process within the Environmental Impact Assessment System in Chile, after ca. 14 years of being enforced. This analysis is sustained by the description and analysis of the stakeholder participation possibilities in a representative rural area of North-Central Chile. The Environmental Basis Act 19300, enacted in 1994, considers the participation of the local community in the environmental impact assessment of new projects. However, this possibility is very limited and difficult to exert, often resulting in frustration for the participants. This is due to a number of reasons, such as the imbalance of resources and knowledge among the majority of participating communities and project proponents, the complexity and administrative and legal constraints to participation, and the dominant interest of the Central Government in approving investments, specifically in energy and natural resources related projects, which generate wealth and jobs. Also, the State's rush to develop Internet-based communication and management systems has built a barrier for poor, traditional communities. This factor is clearly reflected in the case study considered. Results show that there is generally a significant lack of knowledge about institutions and participation tools. From this base, we intend to raise concern on these selected aspects that could be addressed to improve the effectiveness of the existing framework, both in Chile and in other developing countries, where immature environmental impact assessment and public management systems face similar pressures in relation to the sustainable use of their natural resources. Finally, some basic steps are proposed in order to make the community participation an effective tool for sustainable development. PMID- 21641110 TI - Silica materials recovered from photonic industrial waste powder: its extraction, modification, characterization and application. AB - This study explored the possibility of recovering waste powder from photonic industry into two useful resources, sodium fluoride (NaF) and the silica precursor solution. An alkali fusion process was utilized to effectively separate silicate supernatant and the sediment. The obtained sediment contains purified NaF (>90%), which provides further reuse possibility since NaF is widely applied in chemical industry. The supernatant is a valuable silicate source for synthesizing mesoporous silica material such as MCM-41. The MCM-41 produced from the photonic waste powder (PWP), namely MCM-41(PWP), possessed high specific surface areas (1082 m(2)/g), narrow pore size distributions (2.95 nm) and large pore volumes (0.99 cm(3)/g). The amine-modified MCM-41(PWP) was further applied as an adsorbent for the capture of CO(2) greenhouse gas. Breakthrough experiments demonstrated that the tetraethylenepentamine (TEPA) functionalized MCM-41(PWP) exhibited an adsorption capacity (82 mg CO(2)/g adsorbent) of only slightly less than that of the TEPA/MCM-41 manufactured from pure chemical (97 mg CO(2)/g adsorbent), and its capacity is higher than that of TEPA/ZSM-5 zeolite (43 mg CO(2)/g adsorbent). The results revealed both the high potential of resource recovery from the photonic solid waste and the cost-effective application of waste-derived mesoporous adsorbent for environmental protection. PMID- 21641111 TI - Microencapsulation of phosphogypsum into a sulfur polymer matrix: physico chemical and radiological characterization. AB - The aim of this work is to prepare a new type of phosphogypsum-sulfur polymer cements (PG-SPC) to be utilised in the manufacture of building materials. Physico chemical and radiological characterization was performed in phosphogypsum and phosphogypsum-sulfur polymer concretes and modeling of exhalation rates has been also carried out. An optimized mixture of the materials was obtained, the solidified material with optimal mixture (sulfur/phosphogypsum=1:0.9, phosphogypsum dosage=10-40 wt.%) results in highest strength (54-62 MPa) and low total porosity (2.8-6.8%). The activity concentration index (I) in the PG-SPC is lower than the reference value in the most international regulations and; therefore, these cements can be used without radiological restrictions in the manufacture of building materials. Under normal conditions of ventilation, the contribution to the expected radon indoor concentration in a standard room is below the international recommendations, so the building materials studied in this work can be applied to houses built up under normal ventilation conditions. Additionally, and taking into account that the PG is enriched in several natural radionuclides as (226)Ra, the leaching experiments have demonstrated that environmental impact of the using of SPCs cements with PG is negligible. PMID- 21641112 TI - A numerical method to determine the steady state distribution of passive contaminant in generic ventilation systems. AB - Ventilation system with air recirculation is designed to conserve energy, yet at the same time may result in transporting hazardous substance among different rooms in the same building, which is a concern in indoor air quality control. There is a lack of effective methods to predict indoor contaminant distribution primarily because of uncertainty of the contaminant concentration in supply air which in turn due to the mixing ratio of fresh and recirculation air. In this paper, a versatile numerical method to determine the pollutant distribution of ventilation system with recirculation at steady state is proposed based on typical ventilation systems with accessibility of supply air (ASA) and accessibility of contaminant source (ACS). The relationship is established between contaminant concentrations of supply air and return air in a ventilated room or zone. The concentrations of supply air and contaminant distribution in each room can be determined using such parameters as ASA and ACS. The proposed method is validated by both experimental data and numerical simulation result. The computing speed of the proposed method is compared with the iteration method. The comparisons between the proposed method and the lumped parameter model are also conducted. The advantages of the proposed method in terms of accuracy, speed and versatility make it advantageous to be applied in air quality control of complex ventilation systems with recirculation. PMID- 21641114 TI - BTX-A for persistent myofascial TMD pain: interpreting a small clinical trial with negative results. PMID- 21641113 TI - Keratinocyte expression of calcitonin gene-related peptide beta: implications for neuropathic and inflammatory pain mechanisms. AB - Calcitonin gene-related peptide (CGRP) is a vasodilatory peptide that has been detected at high levels in the skin, blood, and cerebrospinal fluid (CSF) under a variety of inflammatory and chronic pain conditions, presumably derived from peptidergic C and Adelta innervation. Herein, CGRP immunolabeling (IL) was detected in epidermal keratinocytes at levels that were especially high and widespread in the skin of humans from locations afflicted with postherpetic neuralgia (PHN) and complex region pain syndrome type 1 (CRPS), of monkeys infected with simian immunodeficiency virus, and of rats subjected to L5/L6 spinal nerve ligation, sciatic nerve chronic constriction, and subcutaneous injection of complete Freund's adjuvant. Increased CGRP-IL was also detected in epidermal keratinocytes of transgenic mice with keratin-14 promoter driven overexpression of noggin, an antagonist to BMP-4 signaling. Transcriptome microarray, quantitative Polymerase Chain Reaction (qPCR), and Western blot analyses using laser-captured mouse epidermis from transgenics, monolayer cultures of human and mouse keratinocytes, and multilayer human keratinocyte organotypic cultures, revealed that keratinocytes express predominantly the beta isoform of CGRP. Cutaneous peptidergic innervation has been shown to express predominantly the alpha isoform of CGRP. Keratinocytes also express the cognate CGRP receptor components, Calcitonin receptor-like receptor (CRLR), Receptor activity-modifying protein 1 (RAMP1), CGRP-receptor component protein (RCP) consistent with known observations that CGRP promotes several functional changes in keratinocytes, including proliferation and cytokine production. Our results indicate that keratinocyte-derived CGRPbeta may modulate epidermal homeostasis through autocrine/paracrine signaling and may contribute to chronic pain under pathological conditions. PMID- 21641115 TI - Fucoidin, a neutrophil rolling inhibitor, reduces damage in a rat electrical burn injury model. AB - BACKGROUND: Electrical injuries induce progressive tissue loss caused by free oxygen radicals released from neutrophil aggregates. Fucoidin, a potent inhibitor of L-selectin function, reduces the aggregation of neutrophils. The aim of this study was to evaluate the effect of fucoidin on tissue damage in rat electrical burn injury model. METHODS: Forty-two male Wistar albino rats (250-300 g) were divided into 3 groups (Group A (n=6), control group without electrical burn injury; Groups B (n=18) and C (n=18), electrical burn injury groups without and with fucoidin therapy, respectively). Three separate analyses were performed at different time points on 6 out of 18 mice from Group B and C at each time point. Biochemistry (myeloperoxidase and malondialdehyde levels) and histopathology (number of neutrophils) of the skin and muscle biopsies at 1st hour; tissue edema (ratio of wet weight/dry weight of extremities) at 24th hour; and necrotic areas at 7th day after electrical injury were evaluated. The electrical burn was induced by exposing rats to 220 V AC between their left upper extremity and right lower extremity for 10 s. Fucoidin was administered as 25 mg/kg intravenous bolus injection at 15 min after electrical burn injury. RESULTS: Myeloperoxidase and malondialdehyde levels, number of neutrophils, tissue edema, and necrotic area were significantly less in fucoidin-applied rats than the group without fucoidin therapy. CONCLUSIONS: Fucoidin inhibits tissue damage induced by electrical burn injury in rats by reducing necrotic area, edema and number of neutrophils. PMID- 21641116 TI - Abnormal regulation of neo-vascularisation in deep partial thickness scalds in rats with diabetes mellitus. AB - OBJECTIVE: This study observed the degree of neo-vascularisation and differential expression of angiogenesis growth factors and their receptor in deep partial thickness scald wound with diabetes mellitus. METHODS: Sixty Sprague-Dawley rats were randomised into a control group and an STZ-induced diabetic group, inflicted with partial-thickness scalding of 20% total body surface area (20% TBSA) on the back. Wound specimens were harvested immediately after scald and on 1, 3, 7, 10, 14 and 21 post-scald days (PSDs) to observe histological changes, and wound healing rates were calculated. The degree of neo-vascularisation in wound (labelled with blue microsphere) and the quantity of vascular endothelial cells (labelled with red CD31) were also measured by double-labelling immunofluorescence. Angiopoietin-1 (Ang-1), Angiopoietin-2 (Ang-2), Tie-2, vascular endothelial growth factor (VEGF) and Flt-1 messenger RNA (mRNA) were analysed by real-time RT-PCR. Ang-1, Ang-2 and VEGF protein expressions were measured by Western blotting. RESULTS: Wound healing was markedly impaired in diabetic rats. The diabetic rats show inhibited vascularity in the wound edge at every time point (the quantitation of vascularity was 60.0+/-3.0 in the control group and 12.0+/-1.4 in the diabetic group, p<0.01 on day 7). Although neo vascularisation in the number of endothelial cells was not significantly different compared with the normal group, part of new vascular endothelial cells did not form the vascular function. After injury, expression of Ang-2 mRNA and protein were increased in both groups, and the normal group showed decreases on day 7, 14 and 21, whereas the diabetic group showed significant increases. Although the expression VEGF and its receptors before injury was higher than the normal group, the level at 1, 3 and 7 days after injury was significantly lower than that 14 days, and that at 21 days after injury was significantly higher than the normal group. CONCLUSION: Vascular endothelial cells can proliferate actively in the diabetic wound with deep partial-thickness burns, but it is still poor in blood supply due to lack of functional capillaries. The mechanism may be related to sustained abnormal high expression of Ang-2 and down-regulated VEGF. PMID- 21641117 TI - Impact of a prenatal cognitive-behavioral stress management intervention on salivary cortisol levels in low-income mothers and their infants. AB - Recent findings suggest that elevated stress levels during the pre- and postpartum period are related to poor maternal and infant health outcomes; yet, few studies have prospectively examined the efficacy of stress management interventions on regulating stress levels among mothers and their infants. The current study examined whether a prenatal cognitive behavioral stress management (CBSM) intervention would be effective in regulating salivary cortisol (a biological marker of stress) and self-reported stress levels among mothers and their infants at six and 18 months postpartum, relative to two control groups. Our sample was comprised of predominantly Spanish-speaking, low-income women (80%; mean age=25+/-5 years) who were screened for depression during their second trimester of pregnancy (M=16+/-5 weeks of gestation). Women at high risk for depression [i.e., having either a past history of major depression or current elevated symptoms of depression (>=16 on CES-D)] were randomized to either a CBSM group (n=24) or a usual care (UC) group (n=33), while a low risk comparison (LRC) group (n=29) was comprised of women not meeting either depression criteria. ANCOVA analyses demonstrated that: (1) infants of women in the CBSM and LRC groups had significantly lower cortisol levels than infants of women in the UC group at six months postpartum (p<.001); and (2) women in the CBSM group had lower cortisol levels than women in the UC group at 18 months postpartum (p<.01). These results suggest that prenatal CBSM interventions may be efficacious in regulating biological markers of stress among mothers and their infants, thereby decreasing their risk for developing health complications over time. PMID- 21641118 TI - Cortisol, sleep, and recovery - Some gender differences but no straight associations. AB - BACKGROUND: Work related fatigue has been suggested as a link in the assumed sequence of events between repeated adverse work demands and the development of work related stress, which may be associated with changes in concentrations of cortisol, psychological overload and, in the long run, health problems. Insufficient sleep is a contributing factor to lack of recovery, but previous studies on associations between subjective aspects of sleep and recovery, and cortisol, have been inconclusive. The aim with the present study was to examine possible associations between cortisol measures and (I) self-rated recovery, (II) occupational fatigue and (III) subjective sleep quality the night preceding cortisol sampling. Further, possible gender differences were tested. METHODS: Salivary cortisol was measured in 581 persons during a working day, at awakening, +30 min and in the evening. Various measures of subjective sleep and recovery were analyzed in relation to cortisol. RESULTS: Few correlations between cortisol and any sleep- or recovery parameters were found. However, some significant associations were found between cortisol and a few measures of more chronic aspects of sleep and recovery. Gender stratified analyses showed somewhat differing associations among men and women. This indicates that possible associations and pathways between lack of recovery/sleepiness and cortisol, and in the long run, unhealth, may not be similar for men and women. PMID- 21641119 TI - P53-binding protein 1: a new player for tumorigenesis and a new target for breast cancer treatment. AB - Breast cancer remains the most common and fatal cancer in women and has been recognized as a genetic disease. Recently, accumulating evidences have showed that p53-binding protein 1 (53BP1) plays an important role in DNA double-strand breaks (DSBs) repair induced by radiation. In vitro experiments have indicated its interaction with many other genes or proteins for tumor suppression or tumorigenesis via pathways associated with DNA repair, cell-cycle control, apoptosis and cell senescence. In vivo studies also showed suppressive effect of 53BP1 on tumor initiation and progression. Therefore, we hypothesize that 53BP1 has a profound effect on suppressing breast cancer as a tumor suppressor and will be an important new biomarker for breast cancer prognosis. Furthermore, 53BP1 gene therapy will be a potential therapeutic strategy for breast cancer. PMID- 21641120 TI - Effect of vitamin A restriction on performance and meat quality in finishing Holstein bulls and steers. AB - Twenty-four Holstein steers and 23 Holstein bulls (initial body weight= 252 +/- 3.5 kg and age= 187 +/- 7.5d) were randomly allocated to 4 treatments arranged in a 2 * 2 factorial design with gender (bulls vs steers) and vitamin A supplementation (either restricted at 1.3 * 1000 IU/kg, VAR, or supplemented at 4.6 * 1000 IU/kg, CTR) to evaluate the effect of vitamin A restriction on performance, carcass and meat quality traits of Holstein steers and bulls. Intramuscular fat was less (P<0.01) in bulls than in steers, and tended (P=0.09) to be greater in VAR than in CTR animals. Oxidative stability tended (P=0.09) to be greater in meat from VAR than from CTR animals at 21 d of ageing. Vitamin A restriction in Holstein bulls does not achieve the same intramuscular fat levels obtained with castration. PMID- 21641121 TI - Effect of housing system, slaughter weight and slaughter strategy on carcass and meat quality, sex organ development and androstenone and skatole levels in Duroc finished entire male pigs. AB - This study aimed at evaluating the effect of housing system (HS), slaughter weight (SW) and strategy (SS) on carcass a nd meat quality, sexual organ development and boar taint in entire males. Twelve pens of 10 pigs were used (two trials). Half of male pens were allowed visual contact with females (MF) and half with males (MM). Half MM or MF were slaughtered at 105 or 130 kg in trial 1, or penwise or by split marketing in trial 2 at 120 kg. Housing system showed no significant effect on carcass or meat quality. MF presented significantly longer testicles and heavier bulbourethral glands compared to MM. The distribution of androstenone and skatole levels was affected by SW but not by HS or SS, samples with androstenone >1 MUg/g of the different groups falling within the range of 16 to 22%. All correlations between androstenone and sex organs were significant. Housing system and slaughter strategy did not reduce the risk of boar tainted carcasses. PMID- 21641122 TI - Delineation of gross tumor volume (GTV) for radiation treatment planning of locally advanced rectal cancer using information from MRI or FDG-PET/CT: a prospective study. AB - PURPOSE: Accurate delineation of target volumes is important to maximize radiation dose to the tumor and minimize it to nontumor tissue. Computed tomography (CT) and magnetic resonance imaging (MRI) are standard imaging modalities in rectal cancer. The aim was to explore whether functional imaging with F-18 fluorodeoxyglucose (FDG) positron emission tomography (PET), combined with CT (FDG-PET/CT) gives additional information to standard pretreatment evaluation and changes the shape and size of the gross tumor volume (GTV). METHODS AND MATERIALS: From 2007 to 2009, 77 consecutive patients with locally advanced rectal cancer were prospectively screened for inclusion in the study at two university hospitals in Sweden, and 68 patients were eligible. Standard GTV was delineated using information from clinical examination, CT, and MRI (GTV MRI). Thereafter, a GTV-PET was defined in the fused PET-CT, and the target volume delineations were compared for total volume, overlap, and mismatch. Pathologic uptake suspect of metastases was also registered. RESULTS: The median volume of GTV-MRI was larger than that of GTV-PET: 111 cm(3) vs. 87 cm(3) (p < 0.001). In many cases, the GTV-MRI contained the GTV defined on the PET/CT images as subvolumes, but when a GTV total was calculated after the addition of GTV-PET to GTV-MRI, the volume increased, with median 11% (range, 0.5-72%). New lesions were seen in 15% of the patients for whom PET/CT was used. CONCLUSIONS: FDG PET/CT facilitates and adds important information to the standard delineation procedure of locally advanced rectal cancer, mostly resulting in a smaller GTV, but a larger total GTV using the union of GTV-MRI and GTV-PET. New lesions were sometimes seen, potentially changing the treatment strategy. PMID- 21641123 TI - Conditional substance abuse and dependence by diagnosis of mood or anxiety disorder or schizophrenia in the U.S. population. AB - BACKGROUND: Little is known about the association of various psychiatric disorders with the risk of developing dependence or abuse among users of various psychoactive substances (conditional dependence, CD; conditional abuse, CA). OBJECTIVES: Evaluate the association of psychiatric disorders with CA only, CD only and CA+CD. METHOD: Secondary analysis of data from 43,093 non institutionalized US adults in the first wave (2001-2002) of the National Epidemiological Survey on Alcohol and Related Conditions. A structured diagnostic interview allowed classification by lifetime psychiatric diagnosis (DSM-IV criteria) and psychoactive substance use. Data were analyzed using weighted proportions, 95% CIs, and weighted logistic regression models to generate odds ratios (OR) adjusted for socio-demographic characteristics. RESULTS: Psychiatric disorders were associated with higher prevalence of psychoactive substance use, regardless of type of disorder or substance. CA, CD and CA+CD prevalence rates were generally higher than unconditional prevalence rates among respondents with and without psychiatric disorders. Respondents with multiple disorders (mainly mood and anxiety disorders) had higher rates of CA+CD on most, but not all, psychoactive substances (e.g., not heroin), while schizophrenia was associated only with higher rates of tranquilizer CA+CD. Psychiatric disorders had few associations with CA only and CD only on psychoactive substances. CONCLUSION: Study findings suggest that mood and anxiety disorders are associated with increased prevalence of substance use and increased transition from use to CA and CD, while schizophrenia is associated with increased transition from abstinence to use, especially for marijuana. Findings did not support the self-medication hypothesis of substance use disorders. PMID- 21641124 TI - Porcine reproductive and respiratory syndrome virus (PRRSV) influences infection dynamics of porcine circovirus type 2 (PCV2) subtypes PCV2a and PCV2b by prolonging PCV2 viremia and shedding. AB - The objective of this study was to determine the effect of porcine reproductive and respiratory syndrome virus (PRRSV) infection on porcine circovirus type 2 (PCV2) subtypes a (PCV2a) or b (PCV2b) viremia and shedding characteristics in oral, nasal and fecal samples in experimentally infected pigs. Twenty-three, 2- to 6-week-old pigs were randomly divided into five groups: negative control (n=3), PCV2a-I (n=5), PCV2a-PRRSV-CoI (n=5), PCV2b-I (n=5), and PCV2b-PRRSV-CoI (n=5). Blood, oral, nasal and fecal swabs were collected in regular intervals from day post inoculation (dpi) 0 until dpi 70 and tested by quantitative real time PCR for the presence and amount of PCV2 DNA and by ELISA for the presence of PCV2-specific antibodies. The results indicate that there were significantly (P<0.05) higher loads of PCV2a and PCV2b DNA in serum, oral swabs, nasal swabs and fecal swabs and a higher prevalence of detectable PCV2 antigen in tissues of pigs concurrently infected with PCV2 and PRRSV compared to pigs singularly infected with PCV2 further confirming that PRRSV enhances replication of PCV2. Moreover, PRRSV infection significantly prolonged the presence of PCV2 DNA in serum and increased the amount of PCV2 DNA in oral and nasal secretions and fecal excretions in the later stages of infection between dpi 28 and 70. Shedding patterns were similar between groups infected with PCV2a and PCV2b, indicating that there was no subtype-specific interaction with the PRRSV isolate used in this study. The results from this study highlight the interaction between PRRSV and PCV2 and the importance of controlling PRRSV infection in order to reduce PCV2 virus loads in pig populations. PMID- 21641125 TI - Goose haemorrhagic hepatitis caused by a new subtype duck hepatitis type 1 virus. AB - Duck hepatitis type 1 virus (DHV-1) causes a fatal disease in ducklings but there is no report of DHV-1 isolation from goose. Recently, cases of a new disease in overfeeding geese were reported from China. The cases were characterized by haemorrhagic hepatitis lesions on liver after post mortem examinations. The flocks showed about 20-40% morbidity and less than 5% mortality. The histopathological lesions showed destroyed structure of hepatocytic tissue, severe vacuolation and necrosis of hepatocytes. Viral antigen could be detected by monoclonal antibody against duck hepatitis type 1 virus (DHV-1) in the cytoplasm of positive hepatocytes. PCR amplified viral sequences with primers specific for recent Korean-like duck hepatitis type 1 virus (DHV-1C). Alignment of the complete sequence demonstrated that the isolated JT strain from goose exhibiting 95.9% identity to DHV-1C AP-03337 strain, and only 75.3% to classical DHV-1 virus. 80% goslings developed haemorrhagic hepatitis after infection with JT strain. This is the first report on the involvement of a DHV-1 virus in goose. PMID- 21641126 TI - Effects of interferon on immediate-early mRNA and protein levels in sensory neuronal cells infected with herpes simplex virus type 1 or pseudorabies virus. AB - Most alphaherpesviruses are able to establish latency in sensory neurons and reactivate upon specific stimuli to cause recurrent symptoms. We have previously shown that interferon (IFN) is capable of inducing a quiescent HSV-1 and PRV infection that strongly resembles in vivo latency in primary cultures of TG neurons. This IFN-induced latency-like quiescence was found to correlate with suppression of the immediate-early protein ICP4 in HSV-1 and its ortholog IE180 in PRV. Here, we mechanistically investigated the IFN-mediated suppression of ICP4 and IE180 in sensory neuronal cells. RT-qPCR showed that mRNA levels of either HSV ICP4 or PRV IE180 at 4 hpi were mildly but not significantly different in IFN-treated samples versus control samples, whereas a strong reduction was observed at 8 hpi and 12 hpi. However, at 4 hpi, HSV ICP4 but not PRV IE180 protein expression was already markedly reduced in IFN-treated samples. In line with this difference in IFN-mediated suppression of HSV ICP4 versus PRV IE180 protein levels, we found that IFN resulted in an increase in phosphorylation of the translation initiation factor eIF2alpha in HSV-infected but not in PRV infected cells. The latter finding indicates that PRV efficiently circumvents IFN mediated translation inhibition by interfering with phosphorylation of eIF2alpha. PMID- 21641127 TI - Ochratoxin A in feed of food-producing animals: an undesirable mycotoxin with health and performance effects. AB - Mycotoxins are secondary fungal metabolites, whose presence in feed- and foodstuffs is unavoidable. Ochratoxin A (OTA) is one of the known mycotoxins with greatest public health and agro-economic significance. Several toxic effects have been ascribed following exposure, namely nephrotoxicity, as well negative impacts in the performance of farm animals, resulting in major economic implications. Of no less importance for the route of human exposure that can also embody the carry over of OTA from feed into animal-derived products is also a concern. For all these reasons the present article updates the worldwide occurrence of OTA in different raw ingredients and finished feed destined to food-producing animals. After that a brief characterization of specie susceptibility and the major rationales is made. An historical overview of field outbreaks linked to OTA exposure in farm animals, concerning the implicated feeds, contamination levels and major clinical and productivity effects is presented. Finally a review of the major animal health and performance potential impacts of animals being reared on contaminated feed is made allied to a perspective regarding its co-occurrence with other mycotoxins, and simultaneous parasitic and bacterial infections. Ultimately, this article aims to be instructive and draw attention to a mycotoxin so often neglected and elapsed from the list of differential diagnosis in farm practice. For the unpredictability and unavoidability of occurrence, OTA will definitely be an enduring problem in animal production. PMID- 21641128 TI - Application of non-structural protein ELISA kits in nationwide FMD surveillance in pigs to demonstrate virus circulation in Taiwan. AB - Large scale surveillance of FMD non-structural protein (NSP) antibody in pigs was conducted to monitor for FMD virus circulation in Taiwan using Ceditest and UBI NSP ELISA kits after recurrence of FMD in 2009. A total of 53,759 serum samples were collected from pigs in the auction markets in 2009. There were 43 farms with positive FMD NSP reactors to both NSP ELISA tests in the nationwide surveillance. After tracing back, clinical examination and the NSP ELISA testing using both Ceditest and UBI on 14 follow-up serum samples from all the herds with confirmed NSP reactors in 2009, there were 4 farms classified as positive on follow-up testing criteria. In this surveillance, we have demonstrated that the NSP ELISA tests of outbreak farms followed by clinical and serological investigation could be used to detect FMD circulation in the pig population in Taiwan even while the national compulsory vaccination program is ongoing. PMID- 21641129 TI - Oxalate-degrading capacities of lactic acid bacteria in canine feces. AB - In this study, lactic acid bacteria in canine feces were isolated and identified, and their oxalate-degrading capacities were evaluated. The oxalate-degrading capacities were determined for 24 of 47 (51.06%) lactic acid bacteria isolates. Of these, 8 isolates [Leuconostoc mesenteroides (RL75), Lactococcus garvieae (CD2), Lactococcus subsp. lactis (CS21), Enterococcus faecium (CL71 and CL72), and Enterococcus faecalis (CD14, CS62, and CD12)] degraded more than 5% of the oxalate present, while the others degraded less than 5% of the oxalate in vitro. Isolates that degraded more than 5% of the oxalate present were selected for further examination. The oxalate-degrading capacities of individual isolates, a mixture of Enterococcus, a mixture of Lactococcus, and a mixture of the eight isolates were evaluated in media containing different concentrations of glucose (sufficient, insufficient, or no glucose). In comparison with the control medium, all of the individual isolates and mixtures of isolates could degrade oxalate in all three groups (P<0.05). In most cases, the isolates growing in medium with 20 g/L of glucose had higher oxalate-degrading capacities than those growing in medium with 2.5 g/L of glucose or no glucose. The mixture of all isolates showed higher oxalate-degrading capacity than the individual isolates and other mixtures. The oxalate-degrading capacities of the isolates were isolate dependent. PMID- 21641130 TI - Urban domestic dog populations as a source of canine distemper virus for wild carnivores in the Coquimbo region of Chile. AB - Urban areas can support dog populations dense enough to maintain canine distemper virus (CDV) and can be a source of infection for rural dogs and free-ranging carnivores. The aim of this study was to investigate the relationships between urban and rural domestic dog and wild carnivore populations and their effects on the epidemiology of CDV to explain retrospectively a CD outbreak in wild foxes in 2003. From 2005 to 2007 a cross-sectional household questionnaire survey was conducted in Coquimbo and Ovalle cities, in three towns and in rural sites along two transects from these cities to the Fray Jorge National Park (FJNP) in the Coquimbo region, Chile. Blood samples were collected from unvaccinated dogs at surveyed households and from free-ranging foxes in rural areas along the transects. The seroprevalence of CDV in domestic dogs was higher in urban than in rural areas and in the later was highest in dogs born before 2001-2002. The seroprevalence of CDV in foxes was higher in areas closer to human settlements. A high seroprevalence in dogs born before 2001-2002 further supports a link between CDV patterns in rural dog and fox populations. In our study area, urban dogs are proposed to be the source of CDV infection to wild carnivores. The large dog population size and density detected in Coquimbo and Ovalle provides optimal conditions for maintaining a large and dense susceptible population of dogs, which can act as a reservoir for highly infectious diseases and could have been the source of infection in the CD outbreak in wild foxes. PMID- 21641131 TI - Epidemiology of Clostridium difficile on a veal farm: prevalence, molecular characterization and tetracycline resistance. AB - Concern has been raised about the potential for Clostridium difficile to be a bovine and foodborne pathogen, yet limited study has been performed in cattle, and none in veal calves. This study evaluated the epidemiology and microbiology of C. difficile on one veal farm. Rectal swabs were obtained from calves within 48 h of arrival and at one, 17 and 21 weeks later. Selective culture for C. difficile was performed. Isolates were characterized by PCR ribotyping and PCR for tcdA, tcdB and cdtA. Tetracycline resistance and resistance genes were investigated. Multivariable logistic regression models were constructed to determine the relationship between shedding of the bacterium and specific ribotypes and the independent variables: time of sampling and area of housing. Calves were twice more likely to test positive 1 week after arrival (51%) when compared to initial results (32%). Shedding at 17 and 21 weeks was significantly lower (2% at both samplings). Ribotype 078 was the most common. Twelve different ribotypes were present initially with only three ribotypes found subsequently. Seventy-six percent (40/53) of isolates initially recovered were tetracycline resistant compared to 93% (81/87) from 2nd sampling. Tetracycline resistance genes were detected in 24% (13/53) of isolates during 1st and in 55% (50/91) during 2nd sampling. The high prevalence of pathogenic C. difficile in veal calves could be of zoonotic concern. The low prevalence before slaughter may be of importance for the evaluation of foodborne risks. Oxytetracycline administration to calves may have an impact on prevalence of C. difficile colonization. PMID- 21641132 TI - Characterization of porcine autism susceptibility candidate 2 as a candidate gene for the number of corpora lutea in pigs. AB - In a previous study, we mapped two quantitative trait loci (QTL) approximately 50cM apart, both influencing the number of corpora lutea in pigs on chromosome 3. One locus included functional candidate genes for proteins related to specific aspects of fertility, such as the follicle-stimulating hormone receptor and the luteinizing hormone/choriogonadotropin receptor. However, specific genes related to the second locus have not yet been identified. This study aims to identify another candidate gene influencing the number of corpora lutea in pigs. Using 12 polymorphic markers, we fine-mapped a narrow region of pig chromosome 3 that had been shown to contain a QTL for corpora lutea. In the critical region, only 1 gene, autism susceptibility candidate 2 (AUTS2), was identified as a positional candidate. Our results demonstrate that the porcine AUTS2 gene consists of 19 exons with a complete open reading frame of 3768bp encoding an AUTS2 protein of 1256 amino acids. We screened the whole coding sequence and parts of the untranslated region for polymorphisms in an F(2) population of Duroc*Meishan crosses. We found 1 ins/del and 7 single nucleotide polymorphisms (SNP), including 2 nonsynonymous variants, c.943C>T in exon 7 and c.2828C>T in exon 19, resulting in P315S and A943V, respectively. The SNP c.943C>T within a proline rich domain was genotyped in several breeds; the C allele occurred in all breeds, whereas the T allele occurred only in Meishan pigs. Using in situ hybridization, the mRNA expression of the AUTS2 gene was observed on granulosa cells in the porcine ovary and thus may be associated with hormone sensitivity. PMID- 21641133 TI - Attenuation of luteolytic response following fish meal supplementation in dairy buffaloes (Bubalus bubalis). AB - Luteolysis of corpus luteum, due to un-inhibited PGF(2alpha) secretion, has been reported to be a cause of early embryonic mortality in dairy animals. The objective of this study was to determine the effects of fish meal (FM) supplementation on the uterine secretion of PGF(2alpha) and hence establish its supplementation as an antiluteolytic strategy in dairy buffaloes. Five cycling Murrah buffaloes were supplemented with 250g FM daily for 55 days in addition to their routine feed and seven buffaloes were kept as non-supplemented control. After 30 days of FM supplementation, the oestrus was synchronized in all the buffaloes using Ovsynch protocol. On day 15 of synchronized cycle, animals were challenged with oxytocin (OT; 100IU) intravenously and blood samples were collected at 15min interval, 1h before to 4h after OT challenge. The PGF(2alpha) response was measured as the venous concentration of 13,14-dihydro-15-keto PGF(2alpha) (PGFM). The mean hourly concentration of PGFM in FM supplemented buffaloes was lower than in the control buffaloes at all the occasions. During peak response (1h post-OT challenge), PGFM concentration was significantly lower (P<0.05) in FM supplemented buffaloes than in the control (197.4+/-41.7pg/ml versus 326.3+/-33.5pg/ml, respectively). Also the percent rise in PGFM after OT challenge in FM supplemented buffaloes was less than the control (11.73% versus 22.47%). The dietary supplementation did not affect the size of corpus luteum (CL) and plasma progesterone concentration. Plasma glucose and total protein concentrations remained within the normal physiological limits during FM supplementation. The present study indicated that supplementing FM decreased the concentrations of PGF(2alpha) without alterations in the size of CL and plasma progesterone concentrations in dairy buffaloes. PMID- 21641134 TI - No mutation but high mRNA expression of Coxsackie-Adenovirus Receptor was observed in both dilated and ischemic cardiomyopathy. AB - The most common causes of acute myocarditis and the possible consequence of dilated cardiomyopathy are virus infections. The receptor of the two most common viruses connected to these myocardial diseases was identified as Coxsackie Adenovirus Receptor. The purpose of this study was to assess Coxsackie-Adenovirus Receptor mRNA expression in the myocardium and search for mutations in the Coxsackie-Adenovirus Receptor gene to compare dilated, inflammatory and ischemic cardiomyopathy with control group. All the myocardial samples were obtained from 35 explanted hearts during heart transplantation, than DNA and RNA were isolated from the muscle samples. cDNA was generated from RNA using reverse transcription, and real-time PCR was performed with relative quantification by beta-actin gene as endogenous control. Using DNA extracted from the myocardial samples, we sequenced all the seven exons of the Coxsackie-Adenovirus Receptor gene. Coxsackie-Adenovirus Receptor mRNA expression was higher in both ischemic and dilated cardiomyopathy groups than in inflammatory cardiomyopathy and healthy control groups. Sequencing of CAR gene showed no sign of mutation. Therefore, the sequences result of CAR exons did not show any mutation or polymorphism, that explains a determinant role of CAR in dilated or ischemic CM. Our results suggest that high mRNA expression of Coxsackie-Adenovirus Receptor may support its role in regeneration of the damaged myocardium rather than having any role in viral mediated heart disease. PMID- 21641135 TI - Safe use of bipolar radiofrequency induced thermotherapy (RFITT) for nasal surgery in patients with cochlear implants. AB - Treatment of nasal turbinate hypertrophy with bipolar radiofrequency-induced thermotherapy (RFITT) is a common indication in patients affected by chronic vasomotor rhinitis. Nonetheless, there are no reports about the safety of such a surgical procedure in cochlear implant (CI) users, due to concerns that the spread of electrical current and heat could damage the CI inner component. We report two cases of CI recipients successfully and safely undergoing RFITT performed by means of the Olympus(r) Celon-ProBreath bipolar coagulation electrode. PMID- 21641136 TI - A new modification of uvulopalatopharyngoplasty for the treatment of obstructive sleep apnea syndrome. AB - OBJECTIVE: To evaluate objective and subjective improvement after applying a new surgical technique, two-piece palatopharyngoplasty (Two-P4), to the treatment of obstructive sleep apnea syndrome (OSAS). METHODS: Twenty-four patients with mild to severe OSAS underwent Two-P4 between January 2002 and November 2007. Polysomnography and Epworth Sleepiness Scale (ESS) score were used to evaluate surgical results. RESULTS: Mean apnea-hypopnea index (AHI) improved from 50.9 to 10.7 after Two-P4. Mean ESS score decreased significantly from 13.0 to 7.7. Body mass index was unchanged after surgery. Objective success as evaluated by a 50% reduction in AHI and by AHI <20 was obtained in 22 of 24 patients (91.7%). Mean reduction in AHI was 76.9% for all 24 patients, 86.2% for patients with Friedman's anatomical stage I, 78.9% for stage II, and 54.5% for stage III. CONCLUSION: Two-P4 is a novel surgical treatment for OSAS patients with a high success rate (91.7%) as evaluated by reductions in AHI. Two-P4 keeps the middle soft palate intact to form independent scars on both sides, which constrict to stretch the soft palate and widen the pharyngeal space. PMID- 21641137 TI - Bacterial biofilms in chronic rhinosinusitis and their relationship with inflammation severity. AB - AIMS: To identify the presence of bacterial biofilms on mucosal specimens from chronic rhinosinusitis (CRS) patients, and evaluate their relationship with severity of CRS. METHODS: A prospective study of biofilms presence on 24 CRS patients compared with 12 controls was designed. The presence of biofilms was determined by scanning electron microscopy (SEM), and associations with the preoperative Lund-MacKay CT scores, Johansson endoscopic scores, and the history of ESS were assessed. RESULTS: Biofilms were found in 13/24 CRS patients (54.2%) but in only 1/12 controls (8.3%; P<0.01). CRS patients with and without biofilms had similar preoperative Lund-MacKay CT and Johansson endoscopic scores (P>0.05). Patients with revision ESS showed a tendency of higher biofilms incidence (5/7, 71.4%) than those undergoing their first procedure (8/17, 47.1%), but did not reach a significant difference (P>0.05). CONCLUSIONS: The higher incidence of biofilms in CRS patients suggests a role in the pathogenesis of CRS, but no correlation with severity of CRS. PMID- 21641138 TI - Inaccurate preoperative imaging assessment on biliary anatomy not increases biliary complications after living donor liver transplantation. AB - BACKGROUNDS AND AIMS: Accurate assessment of graft bile duct is important to plan surgical procedure. Magnetic resonance cholangiopancreatography (MRCP) has become an important diagnostic procedure in evaluation of pancreaticobiliary ductal abnormalities and has been reported as highly accurate. We aim to estimate the efficacy of preoperative MRCP on depicting biliary anatomy in living donor liver transplantation (LDLT), and to determine whether inaccurate preoperative imaging assessment would increase the biliary complications after LDLT. METHODS: The data of 118 cases LDLT were recorded. Information from preoperative MRCP was assessed using intraoperative cholangiography (IOC) as the gold standard. The possible risk factors of recipient biliary complications were analyzed. RESULTS: Of 118 donors, 84 had normal anatomy (type A) and 34 had anatomic variants (19 cases of type B, 9 cases of type C, 1 case of type E, 2 cases of type F and 3 cases of type I) confirmed by IOC. MRCP correctly predicted all 84 normal cases and 17 of 34 variant cases, and showed an accuracy of 85.6% (101/118). The incidence of biliary complications was comparable between cases with accurate and inaccurate classification of biliary tree from MRCP, and between cases with normal and variant anatomy of bile duct. While cases with graft duct opening <=5mm showed a significant higher incidence of total biliary complications (21.1% vs. 6.6%, P=0.028) and biliary stricture (10.5% vs. 1.6%, P=0.041) compared with cases with large duct opening >5mm. CONCLUSION: MRCP could correctly predict normal but not variant biliary anatomy. Inaccurate assessment of biliary anatomy from MRCP not increases the rate of biliary complications, while small-sized graft duct may cause an increase in biliary complications particularly biliary stricture after LDLT. PMID- 21641139 TI - Rapid identification of Lactobacillus plantarum group using the SNaPshot minisequencing assay. AB - This study used SNaPshot minisequencing for species identification within the Lactobacillus plantarum group. A SNaPshot minisequencing assay using dnaK as a target gene was developed, and five SNP primers were designed by analysing the conserved regions of the dnaK sequences. The specificity of the minisequencing assay was evaluated using 35 strains of L. plantarum group species. The results showed that the SNaPshot minisequencing assay was able to unambiguously and simultaneously discriminate strains belonging to the species L. plantarum subsp. plantarum, L. plantarum subsp. argentoratensis, Lactobacillus paraplantarum, Lactobacillus pentosus and Lactobacillus fabifermentans. In conclusion, a rapid, accurate and cost-effective assay was successfully developed for species identification of the members of the L. plantarum group. PMID- 21641140 TI - A case of rectus sheath hematoma due to Foley catheterization after acute urinary retention. AB - Rectus sheath hematoma (RSH) is generally not considered a reason for abdominal pain and its incidence as a cause of abdominal pain is unknown. RSH is a rarely seen but nonetheless an important disease causing abdominal pain. During contractions of the rectus abdominis muscle, the inferior epigastric artery must glide with the muscle to avoid tearing. When the inferior epigastric artery is torn, blood dissects along the rectus sheath leading to hematoma formation. We report a case of spontaneous rectus sheath hematoma due to Foley catheterization after acute urinary retention. In the current case, the rectus abdominis muscle stretched because of overdistended bladder. After the patient was catheterized to drain urinary retention, the rectus abdominis muscle contracted abruptly, and vessels were injured by rapid muscle contraction caused by Foley catheterization. PMID- 21641141 TI - Lipid 20% emulsion ameliorates the symptoms of olanzapine toxicity in a 4-year old. AB - Olanzapine (Zyprexa; Eli Lilly, Indianapolis, IN) is an antipsychotic medication that has been useful in the treatment of psychiatric disorders. In cases of overdose, cardiovascular and neurologic changes are seen. Lipid emulsion has proven successful in relieving the toxicity associated with overdose of lipid soluble drugs. We present a case report of a 4-year-old child who presented with tachycardia and agitation, followed by somnolence after presumed accidental olanzapine ingestion. Treatment with lipid emulsion resulted in amelioration of the symptoms. Inadvertent discontinuation of a lipid emulsion infusion led to recurrence of symptoms, which quickly resolved with an additional loading dose of lipid emulsion. PMID- 21641142 TI - Is America ALSO missing out? The effect of an obstetric resuscitation course on the developing world. PMID- 21641143 TI - Single-dose ziprasidone associated with QT interval prolongation. AB - Ziprasidone has been rarely associated with QT prolongation especially in patients (1) with no underlying cardiac or metabolic disorders, (2) who are receiving no concomitant medications known to prolong the QT interval, and (3) whom therapy is being initiated at a low dose. We report a 47-year-old patient who was agitated with suicidal ideation. He had a history of cocaine use, the last time being 72 hours before emergency department (ED) presentation. His electrocardiogram (ECG) on arrival in the ED showed a QT of 484 milliseconds and a QTc of 475 milliseconds with a pulse of 58 beats per minute. The patient was given 20 mg intramuscular (IM) ziprasidone for agitation. He reported feeling palpitations and weakness 45 minutes after receiving ziprasidone. His QT interval was prolonged on ECG and returned to baseline after 72 hours. Clinicians should consider obtaining an ECG before ziprasidone administration. PMID- 21641144 TI - Purpura fulminans caused by community-associated methicillin-resistant Staphylococcus aureus. AB - Sepsis-induced purpura fulminans is a rare but life-threatening condition characterized by rapidly progressive hemorrhagic infarction of the skin due to dermal vascular thrombosis resulting in tissue loss and severe scarring. Although most commonly related to meningococcal or invasive group A streptococcal disease, it may also be caused by several other bacterial or viral pathogens including Pneumococcus and Varicella. Purpura fulminans associated with Staphylococcus aureus sepsis is rare but has been reported in adults. However, the syndrome is very unusual in children, and to our knowledge, only 2 cases of staphylococcal purpura fulminans have been reported in children, both due to methicillin susceptible S aureus in the United Kingdom. We report the first well-described case of purpura fulminans due to community-associated methicillin-resistant S aureus in a child. PMID- 21641145 TI - External validation of the Glasgow-Blatchford Bleeding Score and the Rockall Score in the US setting. AB - BACKGROUND: The Glasgow-Blatchford Bleeding Score (GBS) and Rockall Score (RS) are clinical decision rules that risk stratify emergency department (ED) patients with upper gastrointestinal bleeding (UGIB). We evaluated GBS and RS to determine the extent to which either score identifies patients with UGIB who could be safely discharged from the ED. METHODS: We reviewed and extracted data from the electronic medical records of consecutive adult patients who presented with signs or symptoms of UGIB (hematemesis and/or melena) to an academic ED from April 1, 2004, to April 1, 2009. The primary outcome was need for intervention (blood transfusion and/or endoscopic/surgical intervention) or death within 30 days. RESULTS: We identified 171 patients with the following characteristics: mean age of 69.9 years (SD, 17.0 years ), 52% women, 20% with a history of liver disease, and 22% with history of gastrointestinal bleeding. Ninety (52.6%, 95% confidence interval, 44.9-60.3) patients had the primary outcome. GBS outperformed pre endoscopy RS [area under the receiver operating characteristic curve (AUC)=0.79 vs 0.62; P=.0001; absolute difference, 0.17]. The prognostic accuracy of GBS and post-endoscopy RS was similarly high (AUC, 0.79 vs 0.72; P=.26; absolute difference, 0.07). The specificity of GBS and RS was suboptimal at all potential decision thresholds. CONCLUSIONS: Although GBS outperformed pre-endoscopy RS, the prognostic accuracy of GBS and post-endoscopy RS was similarly high. The specificity of GBS and RS was insufficient to recommend use of either score in clinical practice. PMID- 21641146 TI - Posterior reversible encephalopathy syndrome presenting as papilledema. AB - Posterior reversible encephalopathy syndrome (PRES) is a central nervous system pathology characterized by headaches, altered mental status, seizures, and visual loss. The syndrome is a clincoradiologic diagnosis, which mandates neuroimaging. The aim of this study is to describe a case of asymptomatic PRES in which the only sign was incidental papilledema found on ophthalmologic examination. A thin 19-year-old female G1P1 s/p natural spontaneous vaginal delivery was referred to our emergency department (ED) by the ophthalmology clinic after finding bilateral papilledema on fundoscopic examination. She denied any fevers, chills, nausea, vomiting, as well as headache, lightheadedness, visual changes, or blurriness. Lumbar puncture was performed, and opening pressure was found to be greater than 55 cmH2O. After collecting Cerebrospinal fluid (CSF) for routine analysis, approximately 15 to 20 mL of CSF was drained. After several revisits to the ED, the neurology clinic was consulted. The magnetic resonance imaging ordered by the neurology clinic, as read by the radiologist, showed a focal lesion in splenium of the corpus callosum and diffusion restriction suggestive of acute infarction (although the anatomical location and age would be unusual). This is an atypical manifestation of PRES. The myriad of clinical symptoms and presentations of PRES has become more identifiable as more case reports of the syndrome are published. This case demonstrates that this atypical syndrome may present in an atypical way. The patient may be asymptomatic, and although imaging defines the diagnosis, a complete physical examination must not be ignored because the only sign may be papilledema. PMID- 21641147 TI - Extracorporeal membrane oxygenation in maternal arrhythmic cardiogenic shock. AB - Very few cases of the use of extracorporeal membrane oxygenation (ECMO) in pregnant women have been reported to date. We report the first case of the use of ECMO for the treatment of cardiac arrhythmia with cardiogenic shock in a pregnant woman. A 28-year-old pregnant woman at 26 weeks of gestation presented with supraventricular tachycardia complicated with cardiogenic shock and fetal distress that was refractory to medication and electrical cardioversion. ECMO was applied, and it facilitated successful radiofrequency ablation. PMID- 21641148 TI - Procedural sedation with propofol: a retrospective review of the experiences of an emergency medicine residency program 2005 to 2010. AB - OBJECTIVES: The objective of this study is to evaluate the types and rates of adverse events associated with the use of propofol for procedural sedation by physicians from our emergency medicine residency program and compare those adverse event rates with those rates already published for all moderate and deep sedatives for procedural sedation, including propofol. METHODS: This study was a retrospective chart review of all 215 procedural sedations performed with propofol in our emergency department (ED) from June 2005 to December 2010. The mean patient age was 29 years (SD, 22.1 years; range, 1-91 years). Adverse events were compiled and examined from chart data and compared with similar published studies on adverse event rates using propofol. RESULTS: Of the 215 patients, 10 (4.65%) experienced adverse events related to procedural sedation with propofol. Our frequency of adverse events was not statistically different from the published rate for all moderate and deep sedatives (P = .407). Of all the adverse events, hypotension was the most common, occurring in 5 (2.33%) of the 215 patients. Of the 215 patients, 3 (1.40%) experienced brief hypoxia, with 2 (0.93%) of 3 patients requiring jaw thrust airway repositioning. Two (0.93%) of the 215 patients developed brief apnea that required brief bag valve mask assisted ventilation. No patient required any advanced airway management. All 215 patients recovered completely from the procedural sedation and were discharged from the ED in stable and improved condition. CONCLUSIONS: The adverse event rates from our study correlate with those of numerous earlier as well as recently published studies of moderate and deep sedatives, including propofol. The disciplined use of propofol by emergency physicians should continue to provide ED patients with the best available management options and care while additional focused and larger scale research is conducted to definitively confirm the premise that emergency physicians can continue to safely perform procedural sedation with propofol. PMID- 21641149 TI - Lodox/Statscan provides benefits in evaluation of gunshot injuries. PMID- 21641150 TI - Acute heart failure registry from high-volume university hospital ED: comparing European and US data. AB - BACKGROUND: Acute heart failure (AHF) is associated with a poor prognosis. OBJECTIVES: The objectives of this study are to describe mechanisms of AHF and to identify the predictors for all-cause mortality by patients admitted for hospitalization by emergency departments (EDs) as well as to compare European and American data. METHODS: We designed a prospective registry of consecutively admitted patients for AHF to a high-volume university hospital ED during a 1-year period (n=202; age, 75+/-11 years; 51% men; ejection fraction, 38%+/-15%). RESULTS: The major causes of AHF were coronary artery disease, often with concomitant mitral regurgitation, hypertension, or atrial fibrillation (>90% of cases). At admission, 24.9% of patients had preserved ejection fractions (>50%); and only 7.7% fulfilled the definition of diastolic AHF. The 30-day and long-term mortality (median follow-up, 793 days) were 20.3% and 31.0%, respectively. A low systolic blood pressure (P=.006), reduced ejection fraction (P=.044), and low serum hemoglobin level (P<.01) emerged as the strongest predictors of all-cause mortality. In patients with AHF without acute myocardial infarction (MI) (63.9%), prescription, at discharge, of statins (P<.05) was independently associated with all-cause mortality. CONCLUSIONS: The patient's blood pressure, ejection fraction, and hemoglobin values, at admission, were identified as the strongest predictors of all-cause mortality. In AHF not triggered by acute MI, long-term use of statins may be associated with reduced survival. The prevalence of diastolic AHF is low. The American AHF population had similar baseline characteristics; needed fewer intensive care unit admissions; had a better 30 days of prognosis, lower incidence of MI, and de novo AHF diagnoses. In a similar subgroup, we observed similar incidences of inotropic support and mechanical ventilation. Our results could not be generalized to all patients with AHF admitted to US EDs. PMID- 21641151 TI - Ondansetron as an effective antiemetic in the rural, out-of-hospital setting. PMID- 21641152 TI - The Emergency Coma Scale as an alternative to the Glasgow Coma Scale. PMID- 21641153 TI - Relevance of sonography for retroperitoneal hematoma. PMID- 21641154 TI - Lower limb myalgias in a man who used to "climb the stairs": an atypical abdominal aortic aneurysm. AB - Atypical presentation of emergency abdominal aortic aneurysm comprises a wide spectrum of symptoms. Lower limbs' involvement is infrequent, usually monolateral and with clear vascular features. We report the case of a 58-year-old patient who complained exclusively about symmetric lower limb myalgias without vascular features, after having repeatedly climbed the stairs of the school he worked in. The surprising final diagnosis was of rupturing abdominal aortic aneurysm; the patient was sent to emergency surgery and survived. PMID- 21641155 TI - Relationship between the hemoglobin level at hospital arrival and post-cardiac arrest neurologic outcome. AB - BACKGROUND: The hemoglobin (Hb) level is an essential determinant of oxygen delivery. The restoration of blood perfusion to vital organs and the capacity for oxygen delivery may be associated with ischemia and reperfusion injuries during cardiac arrest and after cardiac arrest. However, whether the Hb level is associated with neurologic outcome in post-cardiac arrest patients remains unclear. METHODS: Emergency medical service information and clinical demographics were compiled for witnessed out-of-hospital cardiac arrest patients with coma after the restoration of spontaneous circulation. The study end point was defined as a favorable neurologic outcome at 28 days. We evaluated the relationship between the Hb level at the time of hospital arrival and the neurologic outcome using univariate analyses and a multivariate logistic regression analysis. RESULTS: There were 137 witnessed cardiac arrest patients: 49 (35.7%) survived and 34 (24.8%) achieved a favorable neurologic outcome. Univariate analyses showed that the favorable outcome group was characterized as having a higher Hb level, a younger age, a higher percentage of male patients, and ventricular fibrillation as the initial cardiac rhythm. In a multivariate analysis adjusting for potential confounding factors, the Hb level at the time of hospital arrival (odds ratio, 1.26; 95% confidence interval, 1.00-1.58) was an independent predictor of a favorable neurologic outcome. CONCLUSION: A higher Hb level at the time of hospital arrival was associated with a favorable short-term neurologic outcome among post-cardiac arrest patients with a presumed cardiac etiology. PMID- 21641156 TI - Serum chitotriosidase activity in acute appendicitis: preliminary results. AB - Chitotriosidase is one of the most quantitative proteins secreted by activated macrophages, so its activity has been proposed as a biochemical marker of macrophage accumulation. The clinical importance of the chitotriosidase is still largely unknown. Our aim was to evaluate diagnostic accuracy of serum chitotriosidase activity in acute appendicitis (AA). A total of 34 patients with preoperative AA diagnosis (18 men and 16 women; mean age, 28.8+/-10.9 years) were enrolled in this study. The appendix specimens were classified as normal appendix (10 patients) and AA (24 patients). The serum chitotriosidase activity was measured preoperatively. Diagnostic value of the preoperative chitotriosidase activity as assessed through the corresponding receiver operating characteristic curve was well (area under the curve, 0.771; 95% confidence interval, 0.647 0.877; P<.05). Preoperative serum chitotriosidase activity may be a useful marker for diagnosis of AA, and future studies are required to confirm the results presented here. PMID- 21641157 TI - Abnormal urinalysis finding triggered antibiotic prescription for asymptomatic bacteriuria in the ED. PMID- 21641158 TI - Blunt diaphragmatic rupture--a rare but challenging entity in thoracoabdominal trauma. AB - PURPOSE: Delayed diagnosis of blunt traumatic diaphragmatic rupture (BDR) is not uncommon in the emergency department (ED) despite improvement in investigative techniques. We reviewed a large case series of patients diagnosed with blunt traumatic diaphragmatic rupture in order to report demographics, clinical features, and mechanisms of injury of this important but challenging entity. METHODS: From January 2001 through December 2009, 43 patients were diagnosed with BDR at Linkou Chang Gung Memorial Hospital. Demographic data, including sex, age, initial hemodynamic parameters, laboratory data, diagnostic imaging, trauma mechanism, injury location, associated injuries, injury severity score (ISS), time to diagnosis, intensive care unit length of stay (ICU LOS), hospital length of stay (hospital LOS), and mortality, were extracted from hospital records. RESULTS: A total of 43 patients (34 men; 9 women) with BDR were analyzed. Their median age was 37 years (15-82 yrs). Most of these injuries were related to traffic collision (76.8%). The anatomic location of injury to the diaphragm consisted of 24 left-sided (55.8%), 14 right-sided (32.6%),and 5 bilateral diaphragmatic injuries. (11.6%) Hemopneumothorax was the most common associated injury (37.2%). The median diagnostic time was 8 hours (range 2 to 366 hrs). The median ISS score was 18 (range 9 to 41). The median ICU LOS was 4 days (range 0 to 99 ds) and the median HLOS was 19 days (range 1 to 106ds). The total mortality rate was 9.3%. Initial high ISS, initial shock and bilateral diaphragmatic injury significantly increased mortality. CONCLUSION: BDR constitutes a rare entity in thoracoabdominal trauma and most of these injuries were related to traffic collision. High index of suspicion was still the main factor to early diagnosis of this case. The mortality was related to initial shock , bilateral BDR and high ISS. Proper initial resuscitation and correction of other serious injuries may be more life-saving in patients with BDR. PMID- 21641159 TI - Deliberate self-poisoning: factors associated with recurrent self-poisoning. AB - OBJECTIVE: To investigate potential risk factors associated with the repetition of deliberate self-poisoning (DSP). METHODS: Retrospective medical record review of all patients who presented to the emergency department of a tertiary teaching hospital after DSP between January 1, 2000, and December 31, 2009. Repetition of a suicide attempt was determined from reported episodes before the index episode or from 2 visits to the same hospital during the study period. Demographic information, clinical variables, and other variables at the first suicide attempt were investigated for factors associated with repetition of DSP. RESULTS: Of the 967 patients, 203 (21%) presented with repeated suicide attempts. Patients with repeat suicide attempts differed in sex, occupation, living condition, method of DSP, history of psychiatric treatment, reversibility, and psychiatric diagnosis. In the multivariate regression analysis, the only reliable associated factors for repeat suicide attempt were sex (P = .001), living condition (P < .001), method of DSP (P < .036), and history of psychiatric treatment (P < .001). CONCLUSION: Early psychological intervention and close observation is required for patients who are female, living without a family, use antidepressants, and have a history of psychiatric treatment. PMID- 21641160 TI - Do we need echocardiography before commencing thrombolytic therapy also for pulmonary embolism? PMID- 21641161 TI - Cardioprotective effect of glucose-insulin on acute propafenone toxicity in rat. AB - OBJECTIVE: We recently observed a case of propafenone self-poisoning in which the patient was initially unresponsive to conventional therapies such as sodium bicarbonate, dopamine, and norepinephrine but recovered with intravenous glucose insulin infusion. We raised the hypothesis that insulin may have a cardioprotective effect in acute propafenone toxicity. METHODS: We evaluated the effect of glucose-insulin infusion on mortality and electrocardiographic abnormalities during acute propafenone toxicity in rats. After measurements of basal mean arterial pressure, heart rate, PR interval, and QRS duration, rats received intravenous propafenone (36 mg/kg per hour) for 12 minutes. Two minutes after the induction of toxicity, the rats (n=10 per group) received either normal saline solution (NSS) or insulin with glucose. Rats in the insulin-treated (Insulin group) and the NSS-treated (NSS group) groups received an intravenous infusion of 36 mg/kg per hour of propafenone until death occurred. Rats receiving only NSS intravenously without propafenone toxicity served as control (Control group, n=10). RESULTS: Insulin treatment improved survival and delayed the hemodynamic and electrocardiographic consequences of propafenone toxicity. Survival was significantly greater in the insulin group than that in the NSS group (P<.001). Insulin prevented the decline in mean arterial pressure and heart rate (P<.05). Insulin also prevented the increase of the PR interval and the QRS duration (P<.05). CONCLUSION: Glucose-insulin infusion delayed the abnormalities in cardiac conduction and improved rat survival after acute propafenone toxicity. These results suggest a cardioprotective effect of glucose-insulin in acute propafenone toxicity. PMID- 21641162 TI - Noninvasive ventilation use in French out-of-hospital settings: a preliminary national survey. AB - OBJECTIVE: The objective of our study was to describe noninvasive ventilation (NIV) practices (pressure support ventilation and continuous positive airway pressure) in French out-of-hospital mobile intensive care units (SMUR) and their compliance with national consensus guidelines. METHOD: Online inquiry into practices of NIV among managers of French SMUR. Analyzed data include types of NIV devices and use of NIV. RESULTS: Upon 218 SMUR referenced within the SAMU de France database, 118 questionnaires (54%) were processed. Noninvasive ventilation equipment: 91% of the SMUR with at least one type of NIV device (continuous positive airway pressure only=82%; pressure support ventilation only=59%; both=50%) but frequently not with recommended devices. Use of NIV: for acute cardiogenic pulmonary edema, practices were compliant with recommendations in 80% of cases, although there was still room for improvement. For other pathologies, practices were very heterogeneous, not always properly assessed, and frequently not compliant with recommendations. CONCLUSION: To conclude, NIV use in the out of-hospital setting in France seems to meet current recommendations for acute cardiogenic pulmonary edema but not for other pathologies. This could be improved by the use of written procedures, conduction of further studies, and promotion of compliance through education. PMID- 21641163 TI - Rate of intra-abdominal injury after a normal abdominal computed tomographic scan in adults with blunt trauma. AB - OBJECTIVE: The objective of this study is to determine the rate of intra abdominal injury (IAI) in adults with blunt abdominal trauma after a normal abdominal computed tomographic (CT) scan. We hypothesize that the risk of subsequent IAI is so low that hospital admission and observation for possible IAI are unnecessary. METHODS: We conducted a prospective, observational cohort study of adults (>18 years) with blunt trauma who underwent abdominal CT scanning in the emergency department. Computed tomographic scans were obtained with intravenous contrast but no oral contrast. Abnormalities on abdominal CT included all visualized IAIs or any finding suggestive of possible IAI. Patients were followed up to determine the presence or absence of IAI and the need for therapeutic intervention if IAI was identified. RESULTS: Of the 3103 patients undergoing abdominal CT, 2734 (88%) had normal CT scans. The median age was 39 years (interquartile range, 26-51 years); and 2141 (78%) were admitted to the hospital. Eight (0.3%; 95% confidence interval, 0.1%-0.6%) were identified with IAIs after normal abdominal CT scans including the following injuries: pancreas (5), liver (4), gastrointestinal (2), and spleen (2). Five underwent therapy at laparotomy. Abdominal CT had a likelihood ratio (+) of 20.9 (95% confidence interval, 17.7-24.8) and likelihood ratio (-) of 0.034 (0.017-0.068). CONCLUSION: Adult patients with blunt torso trauma and normal abdominal CT scans are at low risk for subsequently identified IAI. Thus, hospitalization for evaluation of possible IAI after a normal abdominal CT scan is unnecessary in most cases. PMID- 21641164 TI - Outcome analysis of cardiac arrest due to hanging injury. AB - OBJECTIVE: The aim of this study was to review patient characteristics and analyze the outcomes in patients who have had cardiac arrest from hanging injuries. METHODS: A retrospective review was performed that examined the victims of out-of-hospital cardiac arrest (OHCA) due to hanging who presented to a tertiary general hospital from January 2000 to December 2009. Utstein style variables were evaluated, and patient outcomes were assessed at the time of hospital discharge using the cerebral performance category (CPC) scale. RESULTS: Fifty-two patients with OHCA due to hanging were enrolled in this study from the aforementioned 10-year inclusion period. Resuscitation attempts were performed in 31 patients (60%), and 21 patients were pronounced dead. In all cases, the first monitored cardiac rhythms were either asystole or pulseless electrical activity (PEA) and were therefore nonshockable rhythms. Of the patients for whom resuscitation was attempted, 13 (42%) experienced a return of spontaneous circulation and 1 revealed cervical spine fracture. Of the 13 return-of spontaneous-circulation patients, 5 survived to be discharged. The mean age of these 5 surviving patients was 36 years. All 5 patients were graded as cerebral performance category 4 at discharge. CONCLUSION: The first monitored cardiac rhythms of patients presenting with OHCA due to hanging were nonshockable rhythms wherein the survival rate of these patients was 9.6%. All of the survivors were relatively young and demonstrated poor neurologic outcomes at discharge. Physicians must consider cervical spine fracture in patients who had cardiac arrest from hanging. PMID- 21641165 TI - Anaphylaxis to black widow spider antivenom. AB - Black widow spider envenomation is commonly reported to poison centers. Black widow spider envenomation produces a clinical syndrome, known as latrodectism, characterized by headache, nausea, vomiting, several muscle cramping and pain, joint stiffness, hypertension, and regional diaphoresis. Black widow spider antivenom (Merck & Co, Inc, West Point, PA USA) is an effective and relatively safe treatment option. There is 1 clear case of anaphylaxis secondary to black widow spider antivenom reported in the medical literature. Here, we report a case of anaphylaxis to antivenom. A 12-year-old boy presented to the emergency department (ED) with diffuse, severe pain 2 1/2 hours after being bitten by a black widow spider on the right lower extremity. In the ED, the patient failed analgesic therapy with fentanyl and was given black widow spider antivenom. Within 45 minutes, he exhibited signs and symptoms consistent with anaphylaxis, including wheezing, chest tightness, pruritus, and urticarial rash. The patient was given standard therapy for anaphylaxis, and all of his signs and symptoms (including the pain secondary to the black widow envenomation) resolved over 6 hours of observation. Leading experts agree that the use of antivenom is indicated in cases of severe envenomation not responsive to standard therapy. Despite concern that the antivenom is an equine-derived whole IgG and can precipitate early hypersensitivity reactions, there is only 1 other reported case of anaphylaxis to the antivenom in the medical literature. PMID- 21641166 TI - Wellness in community living adults: the Weigh to Life program. AB - OBJECTIVE: To examine physiological and health-related quality of life (HRQOL) outcomes in community living adults attending a 12-week combined lifestyle wellness program. METHODS: A sample of overweight and obese adults (n=319) and a subgroup who also had diabetes (n=46 of 319) were studied. The program focuses on dietary, physical activity, and behavioral strategies to promote cardiovascular health. Baseline and 12-week measures were obtained. RESULTS: In the total sample, all physiological and HRQOL outcomes improved (p<.05), except HDL. High attendance was associated with the highest weight loss. In the diabetic subgroup, weight, steps/day, low density lipoprotein, and most aspects of HRQOL improved significantly. CONCLUSION: Physiological and HRQOL benefits can be gained from a 12-week combined lifestyle program; greater benefits were obtained with higher attendance. Although the diabetic subgroup was not large, positive outcomes were realized. PRACTICE IMPLICATIONS: The 12-week combined lifestyle program shows promise for improving outcomes in community living overweight and obese adults who may also be diabetic. By attending class, participants are reminded about strategies they are to apply during the 12-week program and, by program end, they are equipped with a tool kit of strategies for use in everyday life. PMID- 21641167 TI - Introduction: the role of pharmacy in patient education and counseling. PMID- 21641168 TI - Pharmaceutical education in patient counseling: 20h spread over 6 years? AB - OBJECTIVE: To share our experiences with an educational program to increase pharmacy students' patient counseling competency. METHODS: A description is given of the various steps taken to develop an educational program about patient counseling: RESULTS: The Health Belief Model has been chosen as the theoretical model to make students understand patients' drug use behavior. A stepwise development of students' communication skills is combined with assessments in which relatively high norms are applied. Actors act as patients in standardized role plays. In total the basic education consists of 20h of training. CONCLUSION: With a rather small educational investment, 50% of our students achieve the required level of patient counseling. The remaining students succeed to pass the assessments, after having received additional education. PRACTICE IMPLICATIONS: Pharmacy students differ in their needs for educational support in developing their patient counseling competency. These differences are to be taken into account in educational programs addressed to student's patient counseling competency. PMID- 21641169 TI - Successful treatment of recurrent extramammary Paget's disease of the penis and scrotum with imiquimod 5% cream. PMID- 21641170 TI - Hypertension, has treatment a la Carte arrived? PMID- 21641171 TI - Mammary to innominate arteriovenous fistula: endovascular management of an anecdotal complication. AB - An internal thoracic artery pseudoaneurysm associated with an arteriovenous fistula to the innominate vein is a very rare complication after implantation of a cardioverter-defibrillator. We report the successful endovascular management of this unusual complication. PMID- 21641172 TI - Therapeutic hypothermia in acute myocardial infarction: a systematic review. AB - Therapeutic hypothermia preserves neurologic function in post-cardiac arrest patients. Several studies suggest that hypothermia may preserve myocardial function in patients with acute myocardial infarction (AMI). We carried out a systematic review to evaluate the efficacy of therapeutic hypothermia at reducing infarct size and decreasing major adverse cardiac events (MACEs) and all-cause mortality in AMI patients. We searched ClinicalTrials.gov, the Cochrane Clinical Trials Register, EMBASE, and MEDLINE through July 2010 for studies investigating therapeutic hypothermia in AMI patients. Data were extracted by 2 reviewers. We identified 5 studies (693 patients), 2 of which were feasibility trials and 3 of which were randomized controlled trials (RCTs). Feasibility trials showed that therapeutic hypothermia is a safe and feasible intervention. RCTs showed that cardiac outcomes were similar at 30 days' follow-up for the hypothermia and control groups. Mean infarct size ranged from 2.0% to 14.1% of the left ventricle in the hypothermia group and from 8.0% to 13.8% of the left ventricle in the control group. MACEs ranged from 0.0% to 6.2% in the hypothermia group and 3.9% to 10.0% in the control group. All-cause mortality ranged from 0.0% to 3.4% and 2.2% to 10.0% in the hypothermia and control groups, respectively. Subgroup analyses suggested that hypothermia may reduce infarct size in patients with anterior wall infarction. Our review suggests that therapeutic hypothermia is safe and feasible. However, more evidence is needed to determine whether therapeutic hypothermia is associated with improved infarct size, MACEs, or all cause mortality in RCTs of AMI patients. PMID- 21641173 TI - Risk vs benefit for catheter ablation in the small child: when does the Bough break? PMID- 21641174 TI - Hypertension: prevention is the next great challenge and reducing dietary sodium is the starting point. PMID- 21641175 TI - Reduction of delays in primary percutaneous coronary intervention. AB - BACKGROUND: Acute myocardial infarction is a major health issue. Primary percutaneous coronary intervention (PPCI) for ST-elevation myocardial infarction was proved to be superior to fibrinolytic therapy in many randomized trials when done in a timely manner. However, PPCI is associated with delays. Studies have shown that greater delay is associated with increased mortality rate. We applied simple interventions to reduce door-to-balloon time. Our study goal was to evaluate the reduction of delays after our interventions and to monitor 30-day mortality. METHODS: A prospective registry was created to evaluate delays and mortality associated with PPCI. Measures such as annual feedback with suggestions were taken to minimize the delays. Door-to-balloon delays before and after the interventions were compared. RESULTS: A total of 1361 primary PCIs were performed from 2005 to 2008. Of these cases, 1071 patients were transferred from community hospitals. The median door-to-balloon time for transferred patients was 142 minutes for 2005, 138 minutes for 2006, 125 minutes for 2007, and 121 minutes for 2008 (P < 0.001 for 2005 vs. 2008). Door-to-balloon time for patients admitted directly to our centre was 87 minutes in 2005, 74.5 minutes in 2006, 73.5 minutes in 2007, and 74.0 minutes in 2008 (P < 0.001 for 2005 vs. 2008). Thirty-day mortality of these consecutive patients is low (5.2% for 2005 and 3.8% for 2008; P = not significant). CONCLUSIONS: Inexpensive and simple interventions may significantly reduce primary PCI-related delays for transferred cases and patients admitted directly to a centre with PPCI facilities. We also observed a low mortality rate for those consecutive patients. PMID- 21641176 TI - Physiologic evaluation of myocardial bridging: a new analysis for an old disease. AB - BACKGROUND: Myocardial bridging (MB) is the most common congenital coronary anomaly. However, the functional relevance of MB is not well understood. METHODS: Eighteen patients with lone MB were consecutively enrolled. Fractional flow reserve (FFR) was measured before and after dobutamine infusion. Diastolic FFR was calculated by offline analysis. Cutoff values for functional significance of FFR and diastolic FFR were 0.75 and 0.76, respectively. RESULTS: Baseline systolic percent diameter stenosis and lesion length of MB were 70 +/- 16% and 24 +/- 7 mm. FFR and diastolic FFR were 0.92 +/- 0.05 and 0.89 +/- 0.07 at maximal hyperemia induced by adenosine, respectively (P = 0.006). Despite the angiographic stenosis, only 1 lesion was functionally significant. After dobutamine infusion, percent diameter stenosis (84 +/- 11%, P = 0.002) and lesion length (26 +/- 6 mm, P = 0.019) were aggravated and diastolic FFR was lowered (0.84 +/- 0.10, P = 0.006). Two additional lesions became functionally significant after dobutamine infusion. Angiographic percent diameter stenosis at diastole was correlated with dobutamine diastolic FFR (R = -0.58, P = 0.04), but stenosis at systole was not. During median follow-up of 54 months, 2 patients underwent target-lesion revascularization. CONCLUSIONS: Dobutamine increased the morphologic and functional severity of MB. Dobutamine-FFR seems to be helpful in the functional assessment of MB. PMID- 21641177 TI - Canadian Hypertension Education Program: the science supporting New 2011 CHEP recommendations with an emphasis on health advocacy and knowledge translation. AB - This is a summary of the theme, key new recommendations, and supporting science of the 2011 Canadian Hypertension Education Program (CHEP). In 2011, the ACCORD trial challenged current blood pressure treatment targets for people with diabetes. After consideration of multiple factors relating to the ACCORD trial design and its reporting, the current treatment target of <130/80 mm Hg was not changed. A meta-analysis implicated angiotensin receptor blockers in causing cancer; however, weaknesses in the meta-analysis and ongoing close scrutiny of the issue by the U.S. Food and Drug Administration precluded any changes in current CHEP recommendations. New expert opinion-based recommendations were added to assist the management of hypertension in the setting of acute stroke. To promote healthier blood pressure in Canadians, CHEP emphasizes the need for all Canadians-in particular, health care professionals and their organizations-to more actively work with different levels of government to implement healthy public policies. These should build community capacity to promote healthy behaviours with the goal of the prevention of hypertension and its consequences. To aid a substantive knowledge translation gap, health care professionals and people with hypertension can now receive regular CHEP updates by signing up at the Web sites htnupdate.ca and www.myBPsite.ca. PMID- 21641178 TI - Anterior dislocation after a posterior stabilized total knee arthroplasty. AB - Dislocation of a total knee arthroplasty is a rare but serious complication. In previous literature, when dislocation does occur, it is usually in the posterior direction in cases with a posterior stabilized total knee arthroplasty due to cam jump. We report an unusual case of anterior dislocation of an 11-year-old posterior stabilized total knee arthroplasty in a 55-year-old woman with rheumatoid arthritis occurred after a slip. PMID- 21641179 TI - Detection of periprosthetic osteolysis around total knee arthroplasties an in vitro study. AB - Periprosthetic osteolysis is a common cause of revision of total knee arthroplasties (TKAs), with plain anteroposterior and lateral (APL) radiographs being the most common method for screening. The aim of this study was to examine the utility of lesion detection and volume appreciation with APL, paired oblique radiographs, and computed tomography. Defects of different sizes were created in 3 cadaveric knees with a cementless TKA in situ and imaged with APL, oblique, and computed tomography modalities. The resultant images were then shown to 3 arthroplasty surgeons, and the absence or presence of lesions, volume size, and confidence in assessment were recorded. The results suggest that the current practice of APL is inferior for the assessment of periprosthetic osteolysis around TKA. PMID- 21641180 TI - Alterations of the attentional networks in patients with anxiety disorders. AB - Cognitive theories of emotion try to explain how anxious people attend to the world. Despite the increase in empirical research in this field, the specific or general attentional impairments of patients with anxiety disorder is not well defined. We decided to investigate the relationship between pathological anxiety and attentional mechanisms from the broader perspective of the attentional networks. In our study, patients with anxiety disorders and control participants carried out a task to assess efficiency of three attentional networks: orienting, alerting, and executive control. The main result was that anxiety disorders are related to both reduced effectiveness of the executive control network and difficulties in disengaging attention from invalid cues, even when using emotionally neutral information. This relationship between these attentional networks and anxiety may in part explain the problems in the day-to-day functioning of these patients. PMID- 21641181 TI - Critical chronic peripheral arterial disease: does outcome justify crural or pedal bypass surgery in patients with advanced age or with comorbidities? AB - BACKGROUND: We wanted to know the effect of comorbidity, age, and gender on the outcome after surgical below-knee revascularization for critical chronic limb ischemia. METHODS: This is a retrospective study of 624 consecutive patients who underwent below-knee bypass surgery between January 1996 and December 2005 because of chronic peripheral arterial disease (PAD). Patients' characteristics were: diabetes mellitus (DM) n = 445 (71%), coronary artery disease (CAD) n = 310 (49%), dialysis-dependent renal insufficiency (dRI) n = 88 (14%), age >70 years n = 279 (44%), male n = 423 (68%), PAD Fontaine's stage III n = 105 (17%), and PAD stage 4 n =519 (83%). All patients had Trans Atlantic Inter-Society Consensus (TASC) C and D lesions, all were treated with a vein bypass to a crural artery n = 354 (57%) and to a pedal artery n = 270 (43%). Kaplan-Meier analysis and multivariate analysis were performed. RESULTS: The early results were as follows. The 30-day major amputation rate was n = 43 (7%). CAD, dRI, age, and gender did not influence major amputation rate, whereas patients with diabetes had a lower risk of early amputation than those without diabetes. (hazard ratio: 0.49, 95% confidence interval: 0.25-0.95, p < 0.05). The 30-day mortality rate was n = 31 (5%) and was uninfluenced by DM, CAD, and gender. Patients with dRI and octogenarians had a high risk of early death (dRI: 13.6%, octogenarians 9.4%). The late results were as follows. Follow-up rates were: limb salvage n = 596 (95.5%) and survival n = 622 (99.7%). The limb salvage rates at 1, 3, and 5 years were 79.1%, 72.1%, and 66.4%, respectively, and were uninfluenced by DM, CAD, dRI, age, and gender. The mortality rates at 1-, 3-, and 5-years were 79%, 63.4%, and 47.3%, respectively. Comorbidities such as CAD, dRI, and age of >70 years reduced life expectancy significantly. DM did not influence 1, 3 and 5 years of survival. The 5-year survival rates as estimated by Kaplan-Meier analysis after revascularization were: DM, 46%; CAD, 38%; dRI, 19%; and age >70 years, 37%. CONCLUSION: Advanced age and comorbidities reduce life span but not the chance of avoiding major amputation after below-knee bypass surgery for critical limb ischemia. PMID- 21641182 TI - Evaluating the validity of the Nisonger Child Behavior Rating Form--parent version. AB - Youth with intellectual and developmental disabilities (IDD) experience high rates of emotional and behavior problems. The Nisonger Child Behavior Rating Form (NCBRF) is one of the few tools developed to assess these problems in this population. It consists of a 10-item Social Competence section and a 66-item Problem Behavior section. The goal of this study was to examine the factorial, criterion, and convergent validity of the NCBRF. Data from individuals aged five to 18 years (n = 399) were submitted to confirmatory factor analysis (CFA). In addition, scores of different clinical groups (disruptive behavior disorders, mood/anxiety disorders, taking psychotropic medicines) were compared to youth with no significant clinical concerns and associations with Developmental Behavior Checklist (DBC) scores were examined in a subsample of participants. Results of the CFA indicated that the Social Competence subscales better fit the data (RMSEA = .056), than the Problem Behavior subscales (RMSEA = .086). Comparisons across clinical groups and correlations with DBC scores were significant and in the expected directions. This study supports the use of the NCBRF in youth with IDD, though the factor structure of the Problem Behavior section may need to be revisited. PMID- 21641183 TI - Intracardiac echocardiography: a new guiding tool for transcatheter aortic valve replacement. AB - BACKGROUND: Echocardiography has been debated as an adjunct for transcatheter aortic valve replacement (TAVR). The aim of this prospective study was to comparatively evaluate intraprocedural guidance using intracardiac echocardiography (ICE) and transesophageal echocardiography (TEE). METHODS: Fifty high-risk patients with severe aortic stenosis scheduled for TAVR were randomized to either guidance using ICE (group 1; n = 25) or monitoring using TEE (group 2; n = 25). RESULTS: In contrast to TEE, ICE allowed continuous monitoring. The need for probe repositioning during the procedure was much lower in group 1 (0.1 +/- 0.3 vs 5.7 +/- 0.7 maneuvers, P < .001). Compared with TEE, the transcaval intracardiac echocardiographic view provided higher coaxiality with the ascending aorta expressed as the length of ascending aorta depicted (4.9 +/- 1.2 vs 6.1 +/- 1.2 cm, P = .003). Both coronary ostia were more frequently visualized in group 1 (18 vs 2 cases, P < .001). ICE-derived annular measurements were correlated closely with preinterventional readings on TEE (n = 25, r(2) = 0.90, P < .001). TEE underestimated intraprocedural pressure gradients in comparison with preinterventional measurements (mean difference, -10.2 +/- 11.1 mm Hg; n = 11, P = .012), but ICE did not (mean difference, -0.3 +/- 14.1 mm Hg; n = 25, P = .913). ICE and TEE detected newly grown thrombi (2 vs 1 case). Severe complications (e.g., annular dissection, pericardial effusion) were not observed. CONCLUSIONS: ICE, which is compatible with sedation and local anesthesia, can be considered an alternative to TEE for intraprocedural guidance during TAVR. It also seems to match the required work flow during TAVR better than TEE. PMID- 21641184 TI - The left ventricle responds to acute graded elevation of right ventricular afterload by augmentation of twist magnitude and untwist rate. AB - BACKGROUND: The right and left ventricles share the interventricular septum, which mechanically transmits pressure gradients. The aim of this study was to investigate how acute mild or moderate right ventricular (RV) afterload affects left ventricular (LV) function. METHODS: In 14 open-chest pigs (mean weight, 43 +/- 4 kg) with preserved pericardium, acute mild (>35 and <=50 mm Hg) and moderate (>50 and <=60 mm Hg) RV pressure loading conditions were induced by constriction of the pulmonary artery. Hemodynamic parameters and LV twist and untwist were evaluated under each condition. RESULTS: From baseline to mild and moderate RV afterload, the mean RV systolic pressure increased from 31.0 +/- 4.3 to 41.1 +/- 2.7 and 52.7 +/- 3.4 mm Hg (P < .001), while LV twist magnitudes increased from 15.4 +/- 5.1 degrees to 18.5 +/- 3.1 degrees and 19.8 +/- 5.0 degrees (P = .004), respectively. Absolute values of LV untwist rate increased from -116.9 +/- 64.9 degrees /sec to -160.0 +/- 53.3 degrees /sec and -169.1 +/- 47.0 degrees /sec, respectively (P = .001). After adjusting for all variables, only the ratio of the early and atrial components of mitral inflow and RV outflow tract acceleration time was significantly associated with the LV twist magnitude and LV untwist rate. CONCLUSIONS: In an acute setting, the left ventricle responds to suddenly elevated RV afterload and decreased RV stroke volume by promptly increasing its twist magnitude and untwist rate. PMID- 21641185 TI - The recent rise in the frequency of type 1 diabetes: who pulled the trigger? PMID- 21641186 TI - Feeding intervention in cleft lip and palate babies: a practical approach to feeding efficiency and weight gain. AB - Using a disposable syringe to feed 1-14-week-old babies with cleft lip and palate (CLP) was studied. 57 CLP babies were randomly divided into: syringe-fed (intervention) and cup-and-spoon-fed groups and compared with 55 normal breast- or bottle-fed babies. Differences in weight gained from birth to 6, 10 and 14 weeks were compared. Syringe-fed CLP babies fed breast milk had a significant difference in weight gain (0.7 and 0.8 kg) compared with cup-and-spoon-fed babies (0.4 kg), at 10 and 14 weeks, respectively. Normal breast-fed babies gained 0.6 and 0.7 kg. Cup-and-spoon-fed CLP babies fed artificial and breast milk gained 0.5 and 0.6 kg; syringe-fed CLP babies gained 0.6 and 1.2 kg. Normal babies gained 1.0 and 1.7 kg for the same age and food. Average feeding times were 10 ml/1.25 min for syringe-fed and 10 ml/2.08 min for cup-and-spoon-fed CLP babies at 6 weeks. 19 (100%) cup-and-spoon-fed babies exhibited spill and regurgitation at 6 weeks compared with 30 (79%) CLP syringe-fed babies (P<0.05). In both groups spill and regurgitation decreased with age. CLP babies fed with the modified method had a faster feeding time, less spill and regurgitation and gained the same weight as normal babies at 10 and 14 weeks. PMID- 21641187 TI - The effects of hypertension and body mass index on diffusion tensor imaging in schizophrenia. AB - Recently, the negative effects of hypertension and elevated body mass index on cognitive functioning in schizophrenia have been reported (Friedman et al., 2010). Data suggests that cognitive changes in hypertensive patients from the general population may be mediated, in part, by white matter damage. Therefore, we performed diffusion tensor imaging (DTI) in the same subjects studied by Friedman et al. (2010) to investigate the effects of hypertension and elevated body mass index on the fractional anisotropy (FA) of several major white matter tracts. Significant interactions between a diagnosis of schizophrenia and hypertension on FA in several white matter regions were detected. Hypertension was associated with lower FA in the schizophrenic group and higher FA in the same tracts in the non-schizophrenic subjects. These results suggest hypertension induced compensatory mechanisms in the brains of non-schizophrenic patients with hypertension which may be impaired in persons with schizophrenia. PMID- 21641188 TI - Rifampicin-macrolide synergy against Legionella pneumophila serogroup 1 in human macrophages using a quantitative real-time PCR assay. PMID- 21641189 TI - Flip-back, an old trick to face highly contrasted relaxation times: application in the characterization of pharmaceutical mixtures by CPMAS NMR. AB - The (13)C-(1)H CPMAS with flip-back pulse NMR experiment is revisited in view of applications to pharmaceutical mixtures. The analysis of the kinetics of relaxation and CP transfer with and without the flip-back pulse shows that a significant gain in (13)C signal can be expected (thus in experimental time) from the flip-back pulse for protons with long T(1). The gain is of the order of T(1) of the protons expressed in seconds. The experiment is applied on samples with highly contrasted spin-lattice relaxation times T(1) for protons, situation encountered in pharmaceutical mixtures. The application of the flip-back increases significantly the relative signal intensity of the component with the longer T(1), making this component detectable even after using short recycle delays. Therefore, this CPMAS with flip-back experiment could be used routinely to get (13)C CPMAS NMR spectra of mixtures in constant experimental time and signal-to-noise ratio without the need for optimization of the recycle delays, and for whatever may be the degree of crystallinity of the active principal ingredient (API) and/or excipients. PMID- 21641190 TI - Integration of an optical coherence tomography (OCT) system into an examination incubator to facilitate in vivo imaging of cardiovascular development in higher vertebrate embryos under stable physiological conditions. AB - High-resolution in vivo imaging of higher vertebrate embryos over short or long time periods under constant physiological conditions is a technically challenging task for researchers working on cardiovascular development. In chick embryos, for example, various studies have shown that without appropriate maintenance of temperature, as one of the main environmental factors, the embryonic heart rate drops rapidly and often results in an increase in regurgitant flow. Hemodynamic parameters are critical stimuli for cardiovascular development that, for a correct evaluation of their developmental significance, should be documented under physiological conditions. However, previous studies were mostly carried out outside of an incubator or under suboptimal environmental conditions. Here we present, to the best of our knowledge, the first detailed description of an optical coherence tomography (OCT) system integrated into an examination incubator to facilitate real-time in vivo imaging of cardiovascular development under physiological environmental conditions. We demonstrate the suitability of this OCT examination incubator unit for use in cardiovascular development studies by examples of proof of principle experiments. We, furthermore, point out the need for use of examination incubators for physiological OCT examinations by documenting the effects of room climate (22 degrees C) on the performance of the cardiovascular system of chick embryos (HH-stages 16/17). Upon exposure to room climate, chick embryos showed a fast drop in the heart rate and striking changes in the cardiac contraction behaviour and the blood flow through the vitelline circulation. We have documented these changes for the first time by M-mode OCT and Doppler M-mode OCT. PMID- 21641191 TI - MyMiCROscope: intelligent virtual microscopy in a blended learning model at Ulm University. AB - The growing diversity among students and the rapid increase in new technologies entering the system of higher education, demand reconsideration of traditional learning methods. To improve the individual student's learning situation we developed and integrated a novel virtual microscope, MyMiCROscope, into a face-to face approach for teaching microscopic anatomy. The intelligent virtual microscope has not only enabled self-directed learning of the students at their individual learning speed independent of time and place but also offered new possibilities to interact with the user because it implements systematic annotations accessible from different operational levels. Furthermore the alteration of a sole instructor-led course into a blended learning model resulted in a change of the learning behaviour of the students: group work and social interactions were facilitated. The results of this study show the advantages that intelligent virtual microscopy incorporates for self-directed learning and that blended learning in undergraduate medical education is able to fulfil the individual needs of the students and support social interactions without disregarding practical skills. PMID- 21641192 TI - Special issue ancient DNA. PMID- 21641193 TI - Agrobacterium tumefaciens mediated transformation of marine microalgae Schizochytrium. AB - Schizochytrium was a known docosahexaenoic acid producing marine microalgae. In this study, we have developed a novel transformation approach of Schizochytrium using the Agrobacterium tumefaciens (A. tumefaciens) binary vector system. After co-cultivation of Schizochytrium protoplasts with A. tumefaciens harboring pCAMBIA2301 containing the neomycin phosphotransferase II (NPT II) gene as the selectable marker which confers resistance to G418, the Schizochytrium transformants were successfully obtained on the G418-containing plates. The integration and expression of the transgenes were confirmed by PCR analysis and GUS activity assay. To further validate the transformation system, pCAMBIA2301 EGFP containing the egfp gene was introduced into Schizochytrium. The following results demonstrated that the exogenous egfp gene has been successfully incorporated into the genome of Schizochytrium. In addition, the introduced egfp gene expressed efficiently according to the Western blot and fluorescence assay results. More importantly, the majority of the transformants displayed similar biomass and fatty acid production comparing with the wild type strain. Our results demonstrated that exogenous genes could be expressed efficiently in transgenic Schizochytrium, suggesting that genetically engineered Schizochytrium could be explored by this system. PMID- 21641194 TI - Influence of Griffonia simplicifolia on male sexual behavior in rats: behavioral and neurochemical study. AB - The seeds of Griffonia simplicifolia Baill. are rich in 5-HTP (5 hydroxytryptophan), a direct precursor of the neurotransmitter serotonin. In the present study we investigated the influence of the plant extract on male sexual behavior. The seed extract was orally administered to Sprague-Dawley male rats at three dose levels (25, 50 and 100 mg/kg) both acutely and subchronically (daily for 9 days). Mating test with receptive female rats was performed 60 min after the acute treatment or the last dose when repetitively administered. Mount, intromission and ejaculation latencies and post-ejaculatory interval were recorded. Food intake and body weight were measured over the 9-day period of treatment. Microdialysis technique was used to detect the extracellular levels of serotonin (5-HT) and its metabolite 5-hydroxyindoleacetic acid (5-HIAA) in rat brain following the acute administration of the extract dosed at 100mg/kg. The acute treatment significantly increased mount latency (at any dosage), intromission and ejaculation latencies (at 100 mg/kg) and post-ejaculatory interval (at 50 and 100 mg/kg). On the contrary the subchronic treatment failed to exert a significant influence on copulatory behavior. The daily administration of the extract dosed at 50 and 100 mg/kg for 9 days significantly reduced food intake and body weight. Finally in the microdialysis experiments we found a dramatic increase in 5-HT and its metabolite 5-HIAA. PMID- 21641195 TI - Modulation of leukotriene D4 attenuates the development of seizures in mice. AB - The present study has been designed to pharmacologically investigate the effect of Montelukast sodium, a leukotriene D(4) receptor antagonist, and 1,2,3,4, tetrahydroisoquinoline, a leukotriene D(4) synthetic pathway inhibitor, on the pathophysiological progression of seizures using mouse models of kindled epilepsy and status epilepticus induced spontaneous recurrent seizures. Pentylenetetrazole (40 mg kg(-1)) (PTZ) administration every second day for a period of 15 d was used to elicit chemically induced kindled seizure activity in mice. In a separate set of groups, fifty consecutive electroshocks were delivered to mice using corneal electrodes with continuously increasing intensity with an inter-shock interval of 40s. Severity of kindled seizures was assessed in terms of a composite kindled seizure severity score (KSSS). Pilocarpine (100 mg kg(-1)) was injected every twenty minutes until the onset of status epilepticus. A spontaneous recurrent seizure severity score (SRSSS) was recorded as a measure of quantitative assessment of the progressive development of spontaneous recurrent seizures induced after pilocarpine status epilepticus. Sub-acute PTZ administration and electroshock induced the development of severe form of kindled seizures in mice. Severity of kindled seizures was assessed in terms of a composite kindled seizure severity score. Further, pharmacological status epilepticus elicited a progressive evolution of spontaneous recurrent seizures in the animals. However, Montelukast sodium, a leukotriene D(4) receptor antagonist, as well as 1,2,3,4, tetrahydroisoquinoline, a leukotriene D(4) synthetic pathway inhibitor, markedly and dose dependently suppressed the development of kindled seizures as well as pilocarpine induced spontaneous recurrent seizures. Therefore, leukotriene D(4) may be implicated in the pathogenesis of seizures. PMID- 21641196 TI - The influence of Flutter(r)VRP1 components on mucus transport of patients with bronchiectasis. AB - BACKGROUND: The Flutter((r))VRP1 combines high frequency oscillation and positive expiratory pressure (PEP). OBJECTIVE: To separately evaluate the effect of the Flutter((r))VRP1 components (high frequency oscillation and PEP) on mucus transportability in patients with bronchiectasis. METHODS: Eighteen patients with bronchiectasis received sessions with the Flutter((r))VRP1 or PEP for 30 min daily in a randomized, crossover study. The treatment duration was four weeks with one of the therapies, one week of a "wash-out" period and followed by four more weeks with the other treatment. Weekly secretion samples were collected and evaluated for mucociliary relative transport velocity (RTV), displacement in a simulated cough machine (SCM) and contact angle measurement (CAM). For the proposed comparisons, a linear regression model was used with mixed effects with a significance level of 5%. RESULTS: The Flutter((r))VRP1 treatment resulted in greater displacement in SCM and lower CAM when comparing results from the first (9.6 +/- 3.4 cm and 29.4 +/- 5.7 degrees , respectively) and fourth weeks of treatment (12.44 +/- 10.5 cm and 23.28 +/- 6.2 degrees , respectively; p < 0.05). There was no significant difference in the RTV between the treatment weeks for either the Flutter((r))VRP1 or PEP. CONCLUSION: The use of the Flutter((r))VRP1 for four weeks is capable of altering the respiratory secretion transport properties, and this alteration is related to the high frequency oscillation component. PMID- 21641198 TI - Serum oxytocin concentrations in elective caesarean delivery: a randomized comparison of three infusion regimens. AB - BACKGROUND: The aim of this study was to determine serum oxytocin concentrations following different regimens of prophylactic oxytocin administration in women undergoing elective caesarean delivery. METHODS: Thirty healthy pregnant patients were randomized, after clamping of the umbilical cord, to receive intravenous oxytocin in one of the following groups: G1 (n=9), 10 IU of oxytocin infused over 30 min (0.33 IU/min); G2 (n=11), 10 IU of oxytocin infused over 3 min and 45 s (2.67 IU/min); and G3 (n=10), 80 IU of oxytocin infused over 30 min (2.67 IU/min). Both patient and surgeon were blinded to allocation. Uterine tone was assessed by surgical palpation. Serum oxytocin concentration was determined by enzyme immunoassay before anaesthesia (T0) and at 5 (T5), 30 (T30) and 60 (T60) min after the start of oxytocin infusion. RESULTS: Serum oxytocin concentrations (mean+/-standard error, ng/mL) were not significantly different in the groups at T0 (0.06+/-0.02, 0.04+/-0.02 and 0.07+/-0.04, respectively, P=0.76), and T60 (0.65+/-0.26, 0.36+/-0.26 and 0.69+/-0.26, respectively, P=0.58). G3 showed higher concentrations than G1 at T5 (3.65+/-0.74 versus 0.71+/-0.27, P=0.01) and at T30 (6.19+/-1.19 versus 1.17+/-0.37, P<0.01), and were higher than G2 at T30 (6.19+/-1.19 versus 0.41+/-0.2, P<0.01). Haemodynamic data and uterine tone were considered satisfactory and similar in all groups. No additional uterotonic agents were needed. CONCLUSION: Serum oxytocin measurements made using enzyme immunoassay in healthy pregnant women undergoing elective caesarean delivery showed that administration of 80 IU oxytocin over 30 min resulted in higher serum oxytocin levels after 5 and 30 min than the two other regimens. The concentrations did not differ between groups at 60 min. PMID- 21641199 TI - The capacity to consent to epidural analgesia in labour. PMID- 21641197 TI - Balancing ER dynamics: shaping, bending, severing, and mending membranes. AB - The endoplasmic reticulum is a multifunctional organelle composed of functionally and morphologically distinct domains. These include the relatively planar nuclear envelope and the peripheral ER, a network of sheet-like cisternae interconnected with tubules that spread throughout the cytoplasm. The ER is highly dynamic and the shape of its domains as well as their relative content are in constant flux. The multiple forces driving these morphological changes depend on the interaction between the ER and microtubules, membrane fusion and fission events and the action of proteins capable of actively shaping membranes. The interplay between these forces is ultimately responsible for the dynamic morphology of the ER, which in turn is crucial for properly executing the varied functions of this organelle. PMID- 21641200 TI - Intraosseous needles on the delivery suite. PMID- 21641201 TI - Labor and delivery in a patient with hemophilia B. AB - Hemophilia B is a rare X-linked disorder that may cause dramatic bleeding. Women account for only 3.2% of those clinically affected. The X-linked inheritance frequently delays the diagnosis in women and may expose the patient to an increased risk of adverse events. There is limited experience with these patients during labor and delivery. A 28-year-old primiparous woman with hemophilia B (bleeding phenotype) delivered a male infant by an unplanned cesarean delivery under general anesthesia following treatment with factor IX and normalization of her coagulation parameters, guided by thromboelastography. Postpartum vaginal bleeding required transfusion of two units of packed red blood cells. Factor IX supplementation continued for one week. Once diagnosed with hemophilia B, a multidisciplinary approach and advanced antenatal planning can increase the likelihood of a safe delivery. Neuraxial approaches and cesarean delivery are recommended only after normalization of the coagulation profile. The male fetus of a hemophilia A or B patient requires special attention. Operative vaginal delivery and invasive fetal monitoring should be avoided. Thromboelastography is an excellent technique to assess parturients with bleeding disorders or peripartum hemorrhage and may be underused. PMID- 21641202 TI - CMACE 2006-2008. PMID- 21641203 TI - Central venous pressure monitoring in severe preeclampsia: a survey of UK practice. PMID- 21641204 TI - Is it time to abandon complete blinded independent central radiological evaluation of progression in registration trials? PMID- 21641205 TI - Laboratory-scale ultrasound pre-treated digestion of sludge: Heat and energy balance. AB - The objective of this work was to maximize the digestibility of biological sludge to elucidate the feasibility of a new sludge management strategy to recover good quality sludge for agricultural use. The combined effects of organic loading rates (from 0.7 to 2.8g VS L(-1)d(-1)) and the degree of disintegration by anaerobic digestion of sonicated activated sludge were discussed, and the thermal and energetic balances were evaluated. Despite low sonication inputs, sludge digestion performance improved in terms of solids degradation and biogas production depending on the soluble organic load. The biogas production by sonicated sludge was higher (up to 30%) with respect to the control. Filterability improved during digestion of sonicated sludge at medium OLR due to a significant abatement of the fines. Thermal balances indicated that sonication may be a proper system to guarantee self-sustaining WAS mesophilic digestion. Nevertheless, thickening is a pre-requisite to achieve a positive energy balance. PMID- 21641206 TI - Biofuels carbon footprints: Whole-systems optimisation for GHG emissions reduction. AB - A modelling approach for strategic design of ethanol production systems combining lifecycle analysis (LCA) and supply chain optimisation (SCO) can significantly contribute to assess their economic and environmental sustainability and to guide decision makers towards a more conscious implementation of ad hoc farming and processing practices. Most models applications so far have been descriptive in nature; the model proposed in this work is "normative" in that it aims to guide actions towards optimal outcomes (e.g. optimising the nitrogen balance through the whole supply chain). The modelling framework was conceived to steer strategic policies through a geographically specific design process considering economic and environmental criteria. Results shows how a crop management strategy devised from a whole systems perspective can significantly contribute to mitigate global warming even in first generation technologies. PMID- 21641207 TI - New class of acetylcholinesterase inhibitors from the stem bark of Knema laurina and their structural insights. AB - Bioassay-guided extraction of the stem bark of Knema laurina showed the acetylcholinesterase (AChE) inhibitory activity of DCM and hexane fractions. Further repeated column chromatography of hexane and DCM fractions resulted in the isolation and purification of five alkenyl phenol and salicylic acid derivatives. New compounds, (+)-2-hydroxy-6-(10'-hydroxypentadec-8'(E) enyl)benzoic acid (1) and 3-pentadec-10'(Z)-enylphenol (2), along with known 3 heptadec-10'(Z)-enylphenol (3), 2-hydroxy-6-(pentadec-10'(Z)-enyl)benzoic acid (4), and 2-hydroxy-6-(10'(Z)-heptadecenyl)benzoic acid (5) were isolated from the stem bark of this plant. Compounds (1-5) were tested for their acetylcholinesterase inhibitory activity. The structures of these compounds were elucidated by the 1D and 2D NMR spectroscopy, mass spectrometry and chemical derivatizations. Compound 5 showed strong acetylcholinesterase inhibitory activity with IC(50) of 0.573 +/- 0.0260 MUM. Docking studies of compound 5 indicated that the phenolic compound with an elongated side chain could possibly penetrate deep into the active site of the enzyme and arrange itself through pi pi interaction, H-bonding, and hydrophobic contacts with some critical residues along the complex geometry of the active gorge. PMID- 21641208 TI - Efficient synthesis, structural characterization and anti-microbial activity of chiral aryl boronate esters of 1,2-O-isopropylidene-alpha-D-xylofuranose. AB - A simple and efficient synthetic approach toward a series of chiral aryl boronate esters, starting from D-xylose, as anti-microbial agents, is described herein. Minimum inhibitory concentration and zone of inhibition revealed that these derivatives exhibit potent anti-bacterial and anti-fungal properties. Herein, we report the first anti-microbial activity of this class of compounds. All products have been characterized by NMR ((1)H, (13)C and (11)B), IR, elemental and mass spectral study. PMID- 21641209 TI - Renin inhibitors for the treatment of hypertension: design and optimization of a novel series of tertiary alcohol-bearing piperidines. AB - The design and optimization of a novel series of renin inhibitor is described herein. Strategically, by committing the necessary resources to the development of synthetic sequences and scaffolds that were most amenable for late stage structural diversification, even as the focus of the SAR campaign moved from one end of the molecule to another, highly potent renin inhibitors could be rapidly identified and profiled. PMID- 21641210 TI - A discovery of novel Mycobacterium tuberculosis pantothenate synthetase inhibitors based on the molecular mechanism of actinomycin D inhibition. AB - Mycobacterium tuberculosis pantothenate synthetase is a potential anti tuberculosis target, and a high-throughput screening system was previously developed to identify its inhibitors. Using a similar system, we screened a small library of compounds and identified actinomycin D (ActD) as a weak inhibitor of pantothenate synthetase. A new method was established to discover more effective inhibitors by determining the molecular mechanism of ActD inhibition followed by structure-based virtual screening. The molecular interaction of inhibition was determined by circular dichroism and tryptophan fluorescence quenching. The structure-based search and virtual screening were performed using the Molecular Operating Environment (MOE) program and SYBYL 7.5, respectively. Two inhibitors were identified with an IC(50) for pantothenate synthetase that was at least ten times better than that of ActD. PMID- 21641211 TI - Discovery of PH-797804, a highly selective and potent inhibitor of p38 MAP kinase. AB - The synthesis and SAR studies of a novel N-aryl pyridinone class of p38 kinase inhibitors are described. Systematic structural modifications to the HTS lead, 5, led to the identification of (-)-4a as a clinical candidate for the treatment of inflammatory diseases. Additionally, the chiral synthesis and properties of (-) 4a are described. PMID- 21641212 TI - 1-Chloro-2-formyl indenes and tetralenes as antitubercular agents. AB - 1-Chloro-2-formyl indenes and tetralenes have been synthesized using Vilsmeier Haack-Arnold reaction onto indanones and tetralones. Most of these analogues exhibited antitubercular activity against Mycobacterium tuberculosis H37Rv strain with MICs ranging from 30 to 500 MUg/mL. Analogue 13 was further modified to some derivatives. The most active analogue 23 showing MIC at 30 MUg/mL was further evaluated for acute oral toxicity in Swiss albino mice and was found to be safe up to 300 mg/kg dose. PMID- 21641213 TI - Novel purine-based fluoroaryl-1,2,3-triazoles as neuroprotecting agents: synthesis, neuronal cell culture investigations, and CDK5 docking studies. AB - A series of novel purine-based fluoroaryl triazoles were synthesized using the Cu(I) catalyzed 1,3-dipolar cycloaddition reactions (click reactions), and assayed for their neuroprotective effects using fluorescence electron microscopy. Among these triazoles, o-fluorophenylmetyl-triazole, 7, has comparable neuroprotective effect as that of Flavopiridol (1) and Roscovitine (2), the state of the art CDK inhibitors, against the Abeta induced neurotoxicity. These results are substantiated using computer docking methods (DarwinDock/GenDock), which predict that Roscovitine and the triazole 7 bind to the ATP-binding site of CDK5/p25 with comparable binding energies, whereas the corresponding pentafluorophenylmethyl-triazole, 9, has dramatically reduced binding energy (in accordance with its lack of neuroprotection). These combined experimental and theoretical studies support the involvement of CDK5/p25 in the neuronal cell cycle re-entry. PMID- 21641214 TI - Flavonoids as inhibitors of human CD38. AB - CD38 is a multifunctional enzyme which is ubiquitously distributed in mammalian tissues. It is involved in the conversion of NAD(P)(+) into cyclic ADP-ribose, NAADP(+) and ADP-ribose and the role of these metabolites in multiple Ca(2+) signaling pathways makes CD38 a novel potential pharmacological target. The dire paucity of CD38 inhibitors, however, renders the search for new molecular tools highly desirable. We report that human CD38 is inhibited at low micromolar concentrations by flavonoids such as luteolinidin, kuromanin and luteolin (IC(50) <10 MUM). Docking studies provide some clues on the mode of interaction of these molecules with the active site of CD38. PMID- 21641215 TI - Phenyl isoxazole voltage-gated sodium channel blockers: structure and activity relationship. AB - Blocking of certain sodium channels is considered to be an attractive mechanism to treat chronic pain conditions. Phenyl isoxazole carbamate 1 was identified as a potent and selective Na(V)1.7 blocker. Structural analogues of 1, both carbamates, ureas and amides, were proven to be useful in establishing the structure-activity relationship and improving ADME related properties. Amide 24 showed a good overall in vitro profile, that translated well to rat in vivo PK. PMID- 21641216 TI - Discovery of S1P agonists with a dihydronaphthalene scaffold. AB - Structure-activity relationship of sphingosine-1-phosphate receptor agonists was examined. Cinnamyl derivative 1 was modified to improve S1P(1) agonistic activity as well as selectivity over S1P(3) agonistic activity. Dihydronaphthalene derivative 10d was identified as a potent S1P(1) receptor agonist with high selectivity against S1P(3) and enhanced efficacy in lowering peripheral lymphocyte counts in mice. PMID- 21641217 TI - A diazen-1-ium-1,2-diolated nitric oxide donor ester prodrug of 3-(4 hydroxymethylphenyl)-4-(4-methanesulfonylphenyl)-5H-furan-2-one: synthesis, biological evaluation and nitric oxide release studies. AB - A novel hybrid nitric oxide-releasing anti-inflammatory (AI) ester prodrug (NONO coxib 14) wherein an O(2)-acetoxymethyl 1-(2-carboxypyrrolidin-1-yl)diazen-1-ium 1,2-diolate (O(2)-acetoxymethyl PROLI/NO) NO-donor moiety was covalently coupled to the CH(2)OH group of 3-(4-hydroxymethylphenyl)-4-(4-methylsulfonylphenyl)-5H furan-2-one (12), was synthesized. The prodrug 14 released a low amount of NO (4.2%) upon incubation with phosphate buffer (PBS) at pH 7.4 which was significantly higher (34.8% of the theoretical maximal release of two molecules of NO/molecule of the parent hybrid ester prodrug) upon incubation in the presence of rat serum. These incubation studies suggest that both NO and the parent compound 12 would be released from the prodrug 14 upon in vivo cleavage by non-specific serum esterases. The prodrug ester 14 is a selective COX-2 inhibitor that exhibited AI activity (ED(50)=72.2mmol/kg po) between that of the reference drugs celecoxib (ED(50)=30.9MUmol/kg po) and ibuprofen (ED(50)=327MUmol/kg po). The NO donor compound 14 exhibited enhanced inhibition of phenylephrine-induced vasoconstriction of isolated mesenteric arteries compared with that observed under control conditions. These studies indicate hybrid ester AI/NO donor prodrugs (NONO-coxibs) constitutes a plausible drug design concept targeted toward the development of selective COX-2 inhibitory AI drugs that are devoid of adverse cardiovascular effects. PMID- 21641218 TI - Synthesis, transport and antiviral activity of Ala-Ser and Val-Ser prodrugs of cidofovir. AB - We report the synthesis and biological evaluation of Ala-(Val-)l-Ser-CO(2)R prodrugs of 1, where a dipeptide promoiety is conjugated to the P(OH)(2) group of cidofovir (1) via esterification by the Ser side chain hydroxyl group and an ethyl group (4 and 5) or alone (6 and 7). In a murine model, oral administration of 4 or 5 did not significantly increase total cidofovir species in the plasma compared to 1 or 2, but 7 resulted in a 15-fold increase in a rat model and had an in vitro EC(50) value against human cytomegalovirus comparable to 1. Neither 6 nor 7 exhibited toxicity up to 100 MUM in KB or HFF cells. PMID- 21641219 TI - Synthesis of novel triplet drugs with 1,3,5-trioxazatriquinane skeletons and their pharmacologies. 1: Synthesis of triplet drugs with morphinan skeletons. AB - We synthesized symmetrical and nonsymmetrical triplet drugs with 1,3,5 trioxazatriquinane skeletons. The isolation of key intermediates, oxazoline dimers, made it possible to effectively produce nonsymmetrical triplets. Among the synthesized triplets, KNT-93, composed of three identical opioid MU receptor agonists, showed dose-dependent antinociception via the MU receptor. The effect was 56-fold more potent than that of morphine, a representative MU agonist. The profound analgesic effect induced by KNT-93 might result from simultaneous occupation of three MU opioid receptors. PMID- 21641220 TI - Individual patient (n=1) "trials" in Duchenne dystrophy. PMID- 21641222 TI - Development of a scale for "difficulties felt by ICU nurses providing end-of-life care" (DFINE): a survey study. AB - OBJECTIVES: To develop a scale for assessing "difficulties felt by intensive care unit (ICU) nurses providing end-of-life care" (DFINE). DESIGN AND SETTING: A questionnaire survey of nurses in ICUs at general hospitals in the Kanto region, Japan. MAIN OUTCOME MEASURES: The scale was evaluated by exploratory factor analysis, calculation of Cronbach's alpha and test-retest reliability. The Frommelt Attitudes Toward Care of the Dying Scale (FATCOD-B-J) and the Nursing Job Stressor Scale (NJSS) were used to investigate concurrent validity. RESULTS: Respondents were 224 ICU nurses (response rate, 78%) at 18 hospitals. Five factors comprising 28 items were identified, involving difficulties related to: "the purpose of the ICU is recovery and survival"; "nursing system and model nurse for end-of-life care"; "building confidence in end-of-life care"; "caring for patients and families at end-of-life"; and "converting from curative care to end-of-life care". Cronbach's alpha for each factor ranged from 0.61 to 0.8. In terms of test-retest reliability, intraclass correlations for each factor ranged from 0.62 to 0.72. "Building confidence in end-of-life care" in DFINE showed a negative correlation with "positive attitudes towards caring for dying patient" in the FATCOD-B-J (r=-0.4). "Nursing system and model nurse for end-of-life care" in DFINE showed a positive correlation with "conflict with other nursing staffs" (r=0.32) and "conflict with physicians/autonomy" (r=0.31) in the NJSS. CONCLUSION: DFINE demonstrated acceptable reliability and validity. However, additional surveys need to be conducted with a larger sample to further characterise the scale. PMID- 21641223 TI - ICU nurses' perceptions of potential constraints and anticipated support to practice defibrillation: a qualitative study. AB - AIM: The study examines the experience of intensive care nurses in caring for patients in cardiac arrest, and their perceptions of introducing nurse-led defibrillation. METHOD: This was a descriptive, exploratory and qualitative study at an intensive care unit (ICU) of an acute regional hospital in Hong Kong. Twelve registered nurses were purposefully selected for interview. RESULTS: Although all the participants were trained in basic life support, only 50% were trained in advanced cardiac life support (ACLS), and those trained in ACLS described having limited opportunities to apply their defibrillation knowledge. Whilst participants believed that they were theoretically prepared to influence the patient's resuscitation outcomes, newly qualified nurses were reluctant to be accountable for defibrillation. In contrast, experienced nurses were more willing to perform nurse-led defibrillation. Support from management, cooperation between nurses and doctors, regular in-hospital 'real-drill' programmes, sponsorship for training, and the use of alternative defibrillation equipment should be considered to encourage nurse-led defibrillation in ICU settings. CONCLUSION: Nurse-led defibrillation is an approach of delivering prompt care to critically ill patients, and a way ahead for intensive care nursing in Hong Kong. Emphasis on a consistent policy to promote nurse-led defibrillation practice is needed. PMID- 21641225 TI - Butterfly scale form birefringence related to photonics. AB - Wings of the butterflies Morpho aega and Eryphanis reevesi were investigated in the present study by fluorescence, polarization and infra-red (IR) spectroscopic microscopy with the aim of identifying the oriented organization of their components and morphological details of their substructures. These wings were found to exhibit a strong iridescent glow depending on the angle of the incident light; their isolated scales exhibited blue fluorescence. Parallel columns or ridges extend from the pad and sockets to the dented apical scale's region, and they are perpendicular to the ribs that connect the columnar ridges. The scales reveal linear dichroism (LD) visually, when attached on the wing matrix or isolated on slides. The LD was inferred to be textural and positive and was also demonstrated with IR microscopy. The scale columns and ribs are birefringent structures. Images obtained before and after birefringence compensation allowed a detailed study of the scale morphology. Form and intrinsic birefringence findings here estimated and discussed in the context of nonlinear optical properties, bring to the level of morphology the state of molecular order and periodicity of the wing structure. FT-IR absorption peaks were found at wavenumbers which correspond to symmetric and asymmetric (-N-H) stretching, symmetric (-C-H) stretching, amide I (-CO) stretching, amide II(-N-H), and beta-linking. Based on LD results obtained with polarized IR the molecular vibrations of the wing scales of M. aega and E. reevesi are assumed to be oriented with respect to the long axis of these structures. PMID- 21641224 TI - The Arm Profile Score: A new summary index to assess upper limb movement pathology. AB - Although three-dimensional movement analysis is being increasingly used to evaluate upper limb (UL) movements, information on how to interpret the complex data is still missing. This paper introduces a new summary index, the "Arm Profile Score" (APS), to evaluate the severity of UL movement pathology based on kinematic data, similar to the "Gait Profile Score". The APS is calculated from the root mean square (RMS) difference between kinematic data of the individual child with UL movement deficits and average data from typically developing children. The APS can be decomposed into 13 Arm Variable Scores (AVS), representing the different joint angles. The APS, together with the AVSs form the "Arm Movement Analysis Profile" (A-MAP). Face and construct validity were established for eight UL tasks in a group of 20 children with hemiplegic cerebral palsy (HCP). Intra-session variability was low for the different tasks, with median inter-quartile ranges below 2 degrees . Correlation analysis showed few significant correlations between the individual AVSs and between the AVS and APS, implying that the A-MAP provides considerably more information compared to the APS only. The APS also showed good correlations with the House classification, and with measures of muscle tone, manual muscle strength and grip strength. This study provides a sound base to use the APS to evaluate UL movement pathology in children with HCP. Further study will need to confirm its value as an outcome measurement. PMID- 21641226 TI - Nuclear analytical methods: we've got the hammer for your nail. PMID- 21641228 TI - The morphological features and developmental changes of the philtral dimple: a guide to surgical intervention in cases of cleft lip. AB - OBJECTIVE: This study was undertaken to analyse in detail the morphological features of the philtral dimple and the developmental changes of philtral morphology, including a comparison with patients with a cleft lip. PATIENTS AND PARTICIPANTS: Fifty-five normal Japanese adults, seventy-five children and fifteen patients with complete unilateral cleft lip were referred for analysis. DESIGN: 3D facial data were acquired with a non-contacting laser scanner. We calculated the desired linear and angular components, defining the features of the philtrum with analysing software. RESULTS: The philtral dimple was morphologically expressed by the two different deepest points in the horizontal and sagittal sectional view, respectively. These decreased with age and did not show any correlation. In contrast, philtral measurements on the X-Y coordinates increased during the course of development. Most of the linear and angular variables in the patients with the cleft lip had exhibited greater values than the normal subjects, reflecting the deformed philtral morphology. CONCLUSIONS: To obtain an optimally natural philtral construction accompanied by a symmetrical lip in cleft surgery, it is critical to take into consideration both the geometric features of the philtral depth and age-appropriate morphological features of the philtrum as objective criteria. PMID- 21641227 TI - Hydrogen peroxide formation in cacao tissues infected by the hemibiotrophic fungus Moniliophthora perniciosa. AB - In plant-pathogen interaction, the hydrogen peroxide (H2O2) may play a dual role: its accumulation inhibits the growth of biotrophic pathogens, while it could help the infection/colonization process of plant by necrotrophic pathogens. One of the possible pathways of H2O production involves oxalic acid (Oxa) degradation by apoplastic oxalate oxidase. Here, we analyzed the production of H2O2, the presence of calcium oxalate (CaOx) crystals and the content of Oxa and ascorbic acid (Asa)--the main precursor of Oxa in plants--in susceptible and resistant cacao (Theobroma cacao L.) infected by the hemibiotrophic fungus Moniliophthora perniciosa. We also quantified the transcript level of ascorbate peroxidase (Apx), germin-like oxalate oxidase (Glp) and dehydroascorbate reductase (Dhar) by RT-qPCR. We report that the CaOx crystal amount and the H2O2 levels in the two varieties present distinct temporal and genotype-dependent patterns. Susceptible variety accumulated more CaOx crystals than the resistant one, and the dissolution of these crystals occurred in the early infection steps and in the final stage of the disease in the resistant and the susceptible variety, respectively. High expression of the Glp and accumulation of Oxa were observed in the resistant variety. The content of Asa increased in the inoculated susceptible variety, but remained constant in the resistant one. The susceptible variety presented reduced Dhar expression. The role of H2O2 and its formation from Oxa via Apx and Glp in resistant and susceptible variety infected by M. perniciosa were discussed. PMID- 21641229 TI - An easy way to apply orthodontic extraction for impacted lower third molar compressing to the inferior alveolar nerve. AB - PURPOSE: To study the results of an easy orthodontic extraction method for impacted lower third molar removal which had roots compressing to the inferior alveolar nerve (IAN). PATIENTS AND METHODS: Forty patients were divided into two groups according to their desire. Orthodontic traction group (n=20) had brackets or mini bone screws on the antagonist maxillary molars as anchorage for orthodontic traction from 3 to 10 weeks until the roots' tip was away from the IAN, the tooth was then removed. Traditional extraction group (n=20) had the tooth removed immediately by the same surgeon. Post-operative results were compared between the two groups. RESULTS: All 20 patients in the orthodontic extraction group had their lower impacted third molar removed easily without lower lip numbness after surgery, while 5 patients in the traditional extraction group had transient IAN injury and went away 1 week later. There were no anchorage teeth and adjacent mandibular second molar loose or displacement. CONCLUSION: Application of orthodontic brackets or mini bone screws on the antagonist maxillary molars is an easy way for orthodontic extraction of impacted lower third molar with roots' tip compressed to the IAN. It is an effective way to avoid IAN injury during tooth extraction. PMID- 21641230 TI - Optimizing targeted cancer therapy: towards clinical application of systems biology approaches. AB - In cancer, genetic and epigenetic alterations ultimately culminate in discordant activation of signal transduction pathways driving the malignant process. Pharmacological or biological inhibition of such pathways holds significant promise with respect to devising rational therapy for cancer. Thus, technical concepts pursuing robust characterization of kinase activity in tissue samples from cancer patients have been subject of investigation. In the present review we provide a comprehensive overview of these techniques and discuss their advantages and disadvantages for systems biology approaches to identify kinase targets in oncological disease. Recent advances in the development and application of array based peptide-substrate kinase activity screens show great promise in overcoming the discrepancy between the evaluation of aberrant cell signaling in specific malignancies or even individual patients and the currently available ensemble of highly specific targeted treatment strategies. These developments have the potential to result in a more effective selection of kinase inhibitors and thus optimize mechanism-based patient-specific therapeutic strategies. Given the results from current research on the tumor kinome, generating network views on aberrant tumor cell signaling is critical to meet this challenge. PMID- 21641231 TI - Cyclophosphamide-based metronomic chemotherapy: after 10 years of experience, where do we stand and where are we going? AB - Metronomic (low-dose, long-term and frequently administered) chemotherapy has attracted renewed interest for the past few years, in particular because of possible positive association with molecular targeted agents. Cyclophosphamide is the most widely-explored agent in such an approach. The main possible mechanisms of actions identified in preclinical models, whatever the histology of tumor, are the stimulation of the immune system and anti-angiogenic action. Retrospective studies and numerous phase II clinical trials have been published in diverse clinical settings, mainly in patients with highly pretreated advanced tumors. The tolerance seems to be acceptable; some objective responses have been reported. Nevertheless, the regimens were very heterogeneous, and most of these studies are not randomized. This makes it difficult to objectively evaluate the additional value of the metronomic administration of cyclophosphamide. Further clinical trials integrating translational research are necessary to better evaluate the clinical benefit of this promising approach. PMID- 21641232 TI - Failure to diagnose pathology: an avoidable complication in oral and maxillofacial surgery. AB - The routine submission of abnormal tissue for histopathologic diagnosis is a vital link in the appropriate management of patients. Receipt of a biopsy report brings the usual case to its full conclusion. Patients are best served when clinical impressions are verified by histopathologic examination, and this in turn will reduce the likelihood of successful malpractice litigation for failure or delay in diagnosis. PMID- 21641233 TI - Effects of interferon-gamma and granulocyte colony-stimulating factor on antifungal activity of human polymorphonuclear neutrophils against Candida albicans grown as biofilms or planktonic cells. AB - Candida albicans is a leading cause of biofilm-related infections. As Candida biofilms are recalcitrant to host defenses, we sought to determine the effects of interferon-gamma and granulocyte colony-stimulating factor, two pro-inflammatory cytokines, on the antifungal activities of human polymorphonuclear neutrophils (PMNs) against C. albicans biofilms, using an in vitro biofilm model. Priming of PMNs by these cytokines augmented fungal damage of planktonic cells; however, priming of PMNs did not have the same effect against Candida biofilms. Biofilm phenotype appears to play an important role in protecting C. albicans from the innate immune system. PMID- 21641234 TI - Anti-TNF rescue CD4+Foxp3+ regulatory T cells in patients with type 1 diabetes from effects mediated by TNF. AB - The presence of low-grade chronic inflammation is a known feature of long standing diabetes type 1. The association between serum level of several markers of inflammation and severity of DM1 was proven. Serum concentrations of TNF were reported to be elevated in diabetic patients, especially those who developed diabetic complications. Lately, it has been also shown that TNF may impair the subset of naturally arising regulatory T cells, which control autoimmunity. The presented study, for the first time, shows the regulatory T cells in the context of an inflammatory environment that is present in patients with type 1 diabetes. It indicates that TNF reduces the number and frequency of regulatory CD4(+)Foxp3(+) T cells in children with diabetes type 1 and that in vitro treatment with anti-TNF antibody seems to rescue this cell subset from its defective effects. PMID- 21641235 TI - Effect of experimental low back pain on neuromuscular control of the trunk in healthy volunteers and patients with chronic low back pain. AB - Studies of electromyographic (EMG) activity and lumbopelvic rhythm have led to a better understanding of neuromuscular alterations in chronic low back pain (cLBP) patients. Whether these changes reflect adaptations to chronic pain or are induced by acute pain is still unclear. This work aimed to assess the effects of experimental LBP on lumbar erector spinae (LES) EMG activity and lumbopelvic kinematics during a trunk flexion-extension task in healthy volunteers and LBP patients. The contribution of disability to these effects was also examined. Twelve healthy participants and 14 cLBP patients performed flexion-extension tasks in three conditions; control, innocuous heat and noxious heat, applied on the skin over L5 or T7. The results indicated that noxious heat at L5 evoked specific increases in LES activity during static full trunk flexion and extension, irrespective of participants' group. Kinematic data suggested that LBP patients adopted a different movement strategy than controls when noxious heat was applied at the L5 level. Besides, high disability was associated with less kinematic changes when approaching and leaving full flexion. These results indicate that experimental pain can induce neuromechanical alterations in cLBP patients and healthy volunteers, and that higher disability in patients is associated with decreased movement pattern changes. PMID- 21641236 TI - Learning to use novel objects: a training study on the acquisition of novel action representations. AB - Many studies have suggested that the motor system is organized in a hierarchical fashion, around the prototypical end location associated with using objects. However, most studies supporting the hierarchical view have used well-known actions and objects that are highly over-learned. Accordingly, at present it is unclear if the hierarchical principle applies to learning the use of novel objects as well. In the present study we found that when learning to use a novel object subjects acquired an action representation of the end location associated with using the object, as evidenced by slower responses in an action observation task, when the object was presented at an incorrect end location. By showing the importance of knowledge about end locations when learning to use a novel object, the present study suggests that end locations are a fundamental organizing feature of the human motor system. PMID- 21641237 TI - Posterolateral approach for treatment of posterior malleolus fracture of the ankle. AB - Treatment of the posterior malleolus has been debated among orthopedic surgeons. Most orthopedic surgeons will fix the posterior malleolus if it is larger than 25% to 30% of the distal articular surface. The most common method of fixation of the posterior malleolus is by indirect reduction and anteroposterior screws. In the present study, we describe the technique and results of treatment of the posterior malleolus by direct reduction through the posterolateral approach to the ankle. The decision to fix the posterior malleolus was determined by its size and displacement. A total of 12 consecutive patients underwent the posterolateral approach to reduce the posterior malleolus, and these were fixed by posterior plate. Two patients were lost to follow-up in the early postoperative period (both after 2 months). No deep infection or wound dehiscence occurred. Ten patients had adequate (<2-mm displacement of the articular surface) radiologic reduction at the final follow-up visit. There were 2 cases of 2 mm or more of articular surface displacement at the final follow-up visit (1 patient had 2-mm displacement noted in the immediate postoperative period and 1 patient had adequate reduction in the beginning but was displaced with additional follow-up). The posterolateral approach to the ankle is a useful tool to treat certain cases of posterior malleolus fracture. It allows good visualization and stable fixation of the posterior malleolus. PMID- 21641238 TI - Surgical treatment of peripheral aneurysms in patients with Behcet's disease. AB - INTRODUCTION: Our aim was to report our experience with 23 patients presenting with 32 peripheral aneurysms secondary to Behcet's disease (BD) and their outcome after vascular surgery. METHODS: The study was retrospective in nature. Except for those presenting with aneurysm rupture, patients underwent surgery after treatment of acute inflammatory lesions. All aneurysms appeared to be pseudo aneurysms. Graft interposition with polytetrafluoroethylene or saphenous vein was most commonly employed. Postoperatively, all patients were put on immunosuppressive and antiplatelet therapy. Follow-up was done every 6-12 months, complications recorded and managed appropriately. RESULTS: All the patients were males. The mean age at diagnosis of a peripheral aneurysm was 41.0 +/- 9 years. There were 17 (53%) femoral, 8 (25%) popliteal, two carotid, two external iliac, two brachial and one internal iliac aneurysms. Fourteen (61%) patients had a single peripheral aneurysm while nine had two. Surgery was performed for all initially presenting 23 aneurysms. Six patients with multiple peripheral aneurysms had surgery for their second asymptomatic aneurysm. The mean follow-up period was 84 +/- 62 months. Of 29 aneurysms operated on, 7 (24%) anastomotic pseudo-aneurysms and 11 (38%) graft occlusions developed. Five (22%) patients underwent major lower extremity amputations. Six (26%) mortalities were recorded. CONCLUSION: Surgery for peripheral aneurysms in BD is warranted in many instances. Results of operation can be improved by prolonged monitoring. However, despite all efforts, peripheral aneurysm involvement in BD worsens the prognosis. PMID- 21641239 TI - In vivo MRI-based 3D mechanical stress-strain profiles of carotid plaques with juxtaluminal plaque haemorrhage: an exploratory study for the mechanism of subsequent cerebrovascular events. AB - OBJECTIVES: Atherosclerotic plaque features, such as fibrous cap erosion, ulceration and rupture and presence of haemorrhage in carotid plaque are two important characteristics associated with subsequent cerebrovascular events and juxtaluminal haemorrhage/thrombus (JLH/T) indicates these two high-risk characteristics. This study aims to investigate the association between JLH/T and subsequent events in patients suffering from transient ischaemic attack (TIA). Three-dimensional mechanical analysis was employed to represent the critical mechanical stress (P-CStress) and stretch (P-CStretch) within the plaque. METHODS: Fifty TIA patients with mild-to-moderate carotid stenosis (30-69%) underwent high-resolution magnetic resonance imaging (MRI) within 72 h of the acute event and eight were excluded from the analysis due to various reasons. A total of 21 patients were found to have JLH/T in the carotid plaque and 21 did not (N-JLH/T). During a 2-year follow-up period, 11 (52.4%) patients in the JLH/T group experienced recurrent events and none in the N-JLH/T group. Three dimensional plaque structure was reconstructed based on the in vivo MRI for the mechanical analysis. RESULTS: P-CStress of both groups was comparable (N-JLH/T: 174.45 +/- 63.96 kPa vs. JLH/T: 212.60 +/- 89.54 kPa; p = 0.120), but P-CStretch of JLH/T was significantly bigger than that of N-JLH/T (N-JLH/T: 1.21 +/- 0.08 vs. JLH/T: 2.10 +/- 0.53; p < 0.0001). Moreover, there were much bigger variations in stress and stretch of the JLH/T group during one cardiac cycle than in those of N-JLH/T group. CONCLUSIONS: In vivo MRI-depicted JLH/T might be a high risk factor initiating recurrent events, as big deformation appearing around the rupture site might prevent healing and tear the haemorrhage/thrombus away from the host structure and prompt further thrombo-embolic events. PMID- 21641241 TI - Engineering erythrocytes to be erythrosensors: first steps. AB - Molecules can be loaded into mammalian erythrocytes through a reversible lysis pore that forms in the membrane when placed in hypotonic media, the result being resealed red cell ghosts. Many studies on the sidedness of transport processes have utilized this approach. In addition, red cell ghosts encapsulated with enzymes have been used in patients to treat specific enzyme deficiencies, particularly when the substrate can cross the red cell membrane. Our long-term goal is to put fluorescent sensors inside erythrocytes, return the loaded red cell ghosts to the animal or patient, and then monitor the fluorescence non invasively to follow changes in plasma analyte concentration. In this paper, we present a novel dialysis method for making the red cell ghosts. In addition, we present a theoretical analysis showing that it is not necessary that every loaded red cell ghost has the same dye concentration. Finally we discuss the constraints on the optimal affinity for the sensor/analyte interaction. PMID- 21641240 TI - Novel therapeutic candidates, identified by molecular modeling, induce gamma globin gene expression in vivo. AB - The beta-hemoglobinopathies and thalassemias are serious genetic blood disorders affecting the beta-globin chain of hemoglobin A (alpha(2)beta(Alpha)(2)). Their clinical severity can be reduced by enhancing expression of fetal hemoglobin (gamma-globin), producing HbF (alpha(2)gamma(2,)). In studies reported here, gamma-globin induction by 23 novel, structurally-unrelated compounds, which had been predicted through molecular modeling and in silico screening of a 13,000 chemical library, was evaluated in vitro in erythroid progenitors cultured from normal subjects and beta-thalassemia patients, and in vivo in transgenic mice or anemic baboons. Four predicted candidates were found to have high potency, with 4 to 8-fold induction of HbF. Two of these compounds have pharmacokinetic profiles favorable for clinical application. These studies thus effectively identified high potency gamma-globin inducing candidate therapeutics and validated the utility of in silico molecular modeling. PMID- 21641242 TI - Z drug zombies: parasomnia, drug effect or both? PMID- 21641243 TI - Hepatic gene expression and plasma albumin concentration related to outcome after attenuation of a congenital portosystemic shunt in dogs. AB - In dogs with a congenital portosystemic shunt (CPSS), the outcome after CPSS attenuation is difficult to predict but is most likely related to hepatic and vascular proliferation that follows the attenuation. The aim of this study was to evaluate the prognostic value of shunt localization (extrahepatic vs. intrahepatic), plasma albumin concentration and hepatic mRNA expression of 19 genes involved in hepatic and vascular growth. The study population consisted of 48 dogs that were referred for surgical ligation of a single intrahepatic or extrahepatic CPSS. Gene expression was measured in intraoperatively sampled hepatic tissue with quantitative real-time PCR. Albumin, methionine adenosyltransferase 2alpha (MAT2alpha) and HGF activator (HGFac) were positively associated with complete recovery after CPSS attenuation using multivariate statistical analyses. Individual outcome could be correctly predicted in 83% of dogs using albumin, MAT2alpha and HGFac as high or low values compared to cut-off values of 19.5 g/L, 0.457 and 0.974, respectively. These variables predicted outcome after CPSS ligation better than shunt localization or albumin alone. Other evaluated gene products were not correlated with outcome. PMID- 21641244 TI - Effects of essential fatty acid supplementation in dogs with idiopathic epilepsy: a clinical trial. AB - The effects of essential fatty acid supplementation (EFA) on the control of idiopathic epilepsy in dogs were investigated in a blinded, placebo-controlled trial. Fifteen dogs were treated with triple purified Omega-3 oil containing 400 mg eicosapentaenoic acid, 250 mg docosahexaenoic acid and 22 mg vitamin E per 1.5 mL at a dose of 1.5 mL/10 kg once daily for 12 weeks, followed by a 12 week placebo period of supplementation with olive oil. Owners recorded seizure frequency and severity and any adverse events. EFA supplementation did not reduce seizure frequency or severity in dogs with idiopathic epilepsy. PMID- 21641245 TI - Realignment capability of the nCPMG sequence. AB - The nCPMG sequence is based on a particular phase modulation of the refocusing pulse train, and was originally designed for rendering the spin echo amplitude insensitive to the initial magnetization phase. This pulse sequence has the peculiarity of being easily invertible, which enables perfect driven equilibrium experiments, in the absence of relaxation. This magnetization 'realignment' is effective for all three components. Hence the overall operation is transparent. Supporting theory is presented here, together with the first direct experimental proof of the claim. The experiment shows that, with the present stabilization sequence, perfect realignment is indeed made possible for a range of refocusing pulse nutation angles from 130 degrees to 230 degrees . PMID- 21641246 TI - Low-power MRI by Frank-sequence excitation. AB - Recently, a new NMR method employing an rf excitation scheme with strongly reduced power has been introduced, which is based on modulating rf pulses according to Frank sequences. For many applications, a reduction of rf power is essential, e.g. to eliminate bulky rf pre-amplifiers or in medical high-field MRI to preserve patient safety. Another benefit of the new scheme are very short dead times allowing for measurements of samples with short relaxation times. In this work, Frank-sequence excitation is used for low-power imaging for the first time. Results of one-, two-, and three-dimensional imaging experiments are presented and compared to conventional images. PMID- 21641247 TI - Quantitative separation of CEST effect from magnetization transfer and spillover effects by Lorentzian-line-fit analysis of z-spectra. AB - Chemical exchange saturation transfer (CEST) processes in aqueous systems are quantified by evaluation of z-spectra, which are obtained by acquisition of the water proton signal after selective RF presaturation at different frequencies. When saturation experiments are performed in vivo, three effects are contributing: CEST, direct water saturation (spillover), and magnetization transfer (MT) mediated by protons bound to macromolecules and bulk water molecules. To analyze the combined saturation a new analytical model is introduced which is based on the weak-saturation-pulse (WSP) approximation. The model combines three single WSP approaches to a general model function. Simulations demonstrated the benefits and constraints of the model, in particular the capability of the model to reproduce the ideal proton transfer rate (PTR) and the conventional MT rate for moderate spillover effects (up to 50% direct saturation at CEST-resonant irradiation). The method offers access to PTR from z spectra data without further knowledge of the system, but requires precise measurements with dense saturation frequency sampling of z-spectra. PTR is related to physical parameters such as concentration, transfer rates and thereby pH or temperature of tissue, using either exogenous contrast agents (PARACEST, DIACEST) or endogenous agents such as amide protons and -OH protons of small metabolites. PMID- 21641248 TI - Presumed scleral perforation during forced duction testing of the superior oblique muscle. AB - This is a report a case of presumed scleral perforation during globe retropulsion as part of the Guyton exaggerated traction test in a patient with a history of bilateral keratoconus and scleral thinning. Perforation was indicated by the sudden occurrence of hyphema and globe softening. Postoperatively, the hyphema resolved, the intraocular pressure normalized, and the patient regained preoperative visual acuity. PMID- 21641249 TI - Antielevation syndrome with onset of diplopia 10 years after inferior oblique anterior transposition. AB - Antielevation syndrome as a complication of inferior oblique anterior transposition usually appears in the early postoperative period. A 29-year-old woman who had been operated on for right superior oblique palsy developed diplopia 10 years after surgery: motility examination was consistent with an antielevation syndrome. A right inferior oblique recession of 14 mm was performed on the previously transposed muscle; motility improved, and the patient has remained asymptomatic. PMID- 21641250 TI - Pain relief for premature infants during ophthalmology assessment. AB - BACKGROUND: The ophthalmological examination of premature infants, which is essential for the detection of retinopathy of prematurity (ROP), can be painful and distressing for the infant. Various researchers have investigated the benefits of topical anesthesia, oral sucrose, and non pharmacological intervention for pain relief. The purpose of this study is to review the current state of knowledge on the effectiveness of these approaches. METHODS: A literature search was performed with MEDLINE (January 1980 to January 2011) and the Cochrane Central Register of Controlled Trials, Issue 1 of 4 (January 2011), to determine the currently available evidence on methods of pain relief for premature infants undergoing ROP examination. RESULTS: Most studies supported the use of topical proparacaine, which marginally decreased pain without any side effects. Oral sucrose did not significantly reduce pain scores during ROP examinations, and withholding feeding before the examination was not beneficial. Infants given pacifiers had lower pain scores than those without pacifiers, and infants who were nested experienced less distress during and after the procedure. Conflicting data existed on the benefits of different examination techniques, but the insertion of a lid speculum appeared to be the most uncomfortable aspect of the screening examination. CONCLUSIONS: Topical anesthetics marginally reduce pain during eye examination in premature infants. Contrary to standard practice, it appears that patients are more comfortable if they are fed before the examination, and there is no benefit of oral sucrose. Nonpharmacological interventions, including sucking on a pacifier and nesting, may also be beneficial. PMID- 21641251 TI - Adams-Oliver syndrome associated with bilateral anterior polar cataracts and optic disk drusen. AB - Adams-Oliver syndrome (AOS) (MIM 100300) was first described in 1945 as a condition of terminal transverse limb defects and aplasia cutis congenita. Since then, its clinical features have been found to be highly variable and include cardiac defects, abdominal wall defects, vascular malformations, brain abnormalities, and ocular anomalies. We report the case of a 3-year-old girl with AOS who was also found to have bilateral anterior polar cataracts and pseudopapilledema secondary to optic disk drusen. To the best of our knowledge, this is the first case of bilateral anterior polar cataracts and pseudopapilledema secondary to optic disk drusen to be reported in association with the AOS. PMID- 21641252 TI - Quantification of mast cells in the uterine cervix of women infected with human immunodeficiency virus. AB - The objective of the study was to identify mastocytosis in the chorionic epithelium of the uterine cervix in HIV-infected and non-HIV-infected women in autopsy specimens using histochemistry and immunohistochemistry techniques. Sixteen cervical tissue specimens were collected, of which 10 (62.50%) were from HIV-infected women. Histochemical and immunohistochemical techniques were used to evaluate mast cell density using Giemsa stain and anti-mast cell tryptase and anti-mast cell chymase antibodies, respectively. The study of the sheets and counting of mast cells with blue (Giemsa) or brown staining (anti-mast cell tryptase or chymase antibodies) were performed by 3 examiners, and 10 consecutive fields were examined under a light microscope at 400* magnification. A significant difference was found in mast cell density in the chorionic epithelium of the cervix in HIV-infected compared with non-HIV-infected women. The present study may contribute to the characterization of genital mucosa abnormalities and help better understand the potential role of mast cells in HIV-infected women. PMID- 21641253 TI - Diagnostic dilemma: a young woman with Fabry disease symptoms, no family history, and a "sequencing cryptic" alpha-galactosidase a large deletion. AB - Fabry disease, an X-linked lysosomal storage disorder, results from the deficient activity of alpha-galactosidase A (alpha-Gal A). In affected males, the clinical diagnosis is confirmed by the markedly decreased alpha-Gal A activity. However, in female heterozygotes, the alpha-Gal A activity can range from low to normal due to random X-chromosomal inactivation, and diagnostic confirmation requires identification of the family's alpha-Gal A gene mutation. In a young female who had occasional acroparesthesias, corneal opacities, and 15 to 50% of the lower limit of normal leukocyte alpha-Gal A activity, alpha-Gal A sequencing in two expert laboratories did not identify a confirmatory mutation, presenting a diagnostic dilemma. A renal biopsy proved diagnostic and renewed efforts to detect an alpha-Gal A mutation. Subsequent gene dosage analyses identified a large alpha-Gal A deletion confirming her heterozygosity, and she was started on enzyme replacement therapy. Thus, gene dosage analyses can detect large deletions (>50bp) in suspect heterozygotes for X-linked and autosomal dominant diseases that are "sequencing cryptic," resolving molecular diagnostic dilemmas. PMID- 21641254 TI - Neonatal carnitine palmitoyltransferase II deficiency associated with Dandy Walker syndrome and sudden death. AB - Neonatal onset of carnitine palmitoyltransferase II (CPT II) deficiency is an autosomal recessive, often lethal disorder of the mitochondrial beta-oxidation of long-chain fatty acids. It is a rare multiorgan disease which includes hypoketotic hypoglycemia, severe hepatomuscular symptoms, cardiac abnormalities, seizures and lethargy, as well as dysmorphic features. Until now, only 22 affected families have been described in the literature. An increasing number of mutations are being identified in the CPT2 gene, with a distinct genotype phenotype correlation in most cases. Herein we report a new case of neonatal CPT II deficiency associated with Dandy-Walker syndrome and sudden death at 13 days of life. CPT II deficiency was suggested by acylcarnitine analysis of dried-blood on filter paper in the expanded newborn screening. Genetic analysis of the CPT2 gene identified the presence of a previously described mutation in homozygosity (c.534_558del25bpinsT). All lethal neonatal CPT II deficiency patients previously described presented severe symptoms during the first week of life, although this was not the case in our patient, who remained stable and without apparent vital risk during the first 11 days of life. The introduction of tandem mass spectrometry to newborn screening has substantially improved our ability to detect metabolic diseases in the newborn period. This case illustrates the value of expanded newborn screening in a neonate with an unusual clinical presentation, combining hydrocephalus and sudden death, that might not commonly lead to the suspicion of an inborn error of metabolism. PMID- 21641255 TI - A case report of an ectopic fetus in a cat. AB - An ectopic fetus was discovered in an 18-month-old uniparous queen that was admitted for an elective ovariectomy. Six months prior she had delivered three healthy kittens. During the preoperative examination, a mass similar in size to a full-term fetus was detected in the abdominal cavity. Ultrasound examination revealed the mass to be an ectopic fetus in the mid-abdominal region. A mummified fetus was removed by laparotomy. No rupture of the uterine wall was visible, but a small necrotic area was present on the left uterine horn, adjacent to the very proximal portion of the uterine horn. The fetus, which was fully developed and covered by a thin membrane, was carefully dissected. Histological examination did not enable us to definitively prove the extra-uterine development of the fetus; however, the ectopic development of the conceptus secondarily expelled into the peritoneal cavity could be assumed. PMID- 21641256 TI - [Linezolid-associated severe metabolic acidosis and anaemia]. PMID- 21641257 TI - [Platelet aggregation disorder due to amitriptyline]. PMID- 21641258 TI - [A descriptive study of oseltamivir-treated hospitalised patients]. PMID- 21641259 TI - [Laronidase for treating post-surgical respiratory failure in a patient with type I mucopolysaccharidosis]. PMID- 21641260 TI - Corynebacterium diphtheriae endocarditis: a case series and review of the treatment approach. AB - OBJECTIVES: Infective endocarditis due to non-toxigenic Corynebacterium diphtheriae is uncommon; we report 10 cases occurring over a 14-year period in Auckland, New Zealand and review the approach for treatment. CASE SERIES: Eight of the 10 patients had known prosthetic valves or homografts in situ. Three patients required surgical intervention for infective endocarditis. Seven patients were treated with a combination of beta-lactam and aminoglycoside, and one each was treated with a combination of vancomycin and an aminoglycoside, a beta-lactam alone, and vancomycin alone. All patients survived and none relapsed. REVIEW OF LITERATURE: The antibiotic treatment of 46 previously reported cases was reviewed; patients treated with a beta-lactam and aminoglycoside (n=25), and without the addition of an aminoglycoside (n=11) were compared. The differences in length of treatment within each group make the comparison of outcome (mortality, need for surgical intervention, disease and treatment complications) difficult. However, regardless of the length of treatment, there was no difference in mortality or need for surgical intervention between the two groups in the currently published cases. CONCLUSIONS: Current evidence suggests that endocarditis of either native or prosthetic valves, caused by penicillin susceptible C. diphtheriae, demonstrates a favorable outcome when treated with either a beta-lactam alone or in combination with an aminoglycoside. Patient specific factors will determine which approach is more appropriate for each individual patient. PMID- 21641261 TI - First case of Legionella pneumophila native valve endocarditis. AB - We report the first case in the English language literature, to our knowledge, of native valve endocarditis due to Legionella pneumophila. The patient had no prior history of cardiothoracic intervention or congenital valvular process. A transesophageal echocardiogram showed a vegetation on the aortic valve. Blood culture and bronchoalveolar lavage returned positive for L. pneumophila. The patient was treated with levofloxacin for 6 weeks total after a second set of blood cultures were negative. The patient survived a complicated hospital course and was discharged to a rehabilitation facility. PMID- 21641262 TI - Improved aorto-ventricular matching in ischemic dilated cardiomyopathy patients after surgical ventricular restoration. AB - SCOPE: This paper contains (i) derivation of the aorto-ventricular matching (AVM) index in terms of the ratio of aortic elastance and LV end-systolic elastance, E(aorta)/E(es); (ii) procedure for determination of this index, by means of non invasive measurements of auscultatory pressures, time-variation of blood volume ejected into the aorta, stroke volume and ejection fraction; (iii) results of improved AVM index evaluation in ischemic dilated cardiomyopathy (IDCM) patients following surgical ventricular restoration (SVR), as a result of reduced end diastolic and end-systolic LV volumes and increased LV E(es). METHODOLOGY: Among the ten recruited IDCM patients, four of them underwent surgical ventricular restoration (SVR) and coronary artery bypass graft (CABG), while six of them underwent CABG alone. All patients were studied by echocardiography pre- and 4 months post-operatively; LV volumes were determined by echo Doppler. LV end systolic elastance E(es) was determined from a derived expression, by employing blood pressure, stroke volume, ejection fraction, pre-ejection and systolic periods, and estimated normalized ventricular elastance at end-diastole, based on single-beat measurements. Aortic elastance E(aorta) was determined by means of our modified single-beat method for determining aortic pressure profile. RESULTS: In the CABG plus SVR group, the AVM index E(aorta)/E(es) was reduced by 35% from 0.93+/-0.32 to 0.60+/-0.33, consistent with improved aorto-ventricular matching. However, in the CABG alone group, the AVM index E(aorta)/E(es) decreased only 11% from 1.02+/-0.24 to 0.91+/-0.29. CONCLUSION: There is shown to be increased value of LV E(es) and a more favorable decreased value of AVM index in those IDCM patients who underwent SVR. PMID- 21641263 TI - Trunk accelerometry reveals postural instability in untreated Parkinson's disease. AB - While several studies have shown that subjects with advanced Parkinson's disease (PD) exhibit abnormalities in sway parameters during quiet standing, abnormalities of postural sway associated with untreated PD have not been reported. Although not clinically apparent, we hypothesized that spontaneous sway in quiet stance is abnormal in people with untreated PD. We examined 13 subjects, recently diagnosed with PD, who were not yet taking any anti-parkinsonian medications and 12 healthy, age-matched control subjects. Postural sway was measured with a linear accelerometer on the posterior trunk (L5 level) and compared with traditional force plate measures of sway. Subjects stood for 2 min under two conditions: eyes open (EO) and eyes closed (EC). One of the most discriminative measures of postural changes in subjects with untreated PD was the increased 'JERK' of lower trunk in the EO condition, measured with the accelerometer. Root mean square and the frequency dispersion of postural sway in the EO condition also discriminated sway in untreated PD subjects compared to control subjects. We conclude that accelerometer-based sway metrics could be used as objective measures of postural instability in untreated PD. Accelerometer based analysis of spontaneous sway may provide a powerful tool for early clinical trials and for monitoring the effects of treatment of balance disorders in subjects with PD. PMID- 21641264 TI - Safety of transcranial magnetic stimulation for the newer generation of deep brain stimulators. PMID- 21641265 TI - Prognostic significance of neurologic examination findings in Wilson disease. AB - BACKGROUND: Wilson disease patients present with any of several neurologic phenotypes, and their treated outcomes vary widely. Our goal was to determine whether presenting clinical features of neurologic Wilson disease (WD) predict longer term neurologic outcomes in patients receiving anticopper treatment. METHODS: Patients enrolled in four WD treatment trials received a standardized neurologic examination at trial enrollment and then at pre-specified intervals following anticopper therapy, initially with tetrathiomolybdate or trientine and then with zinc. The examination scored patients' motor signs, including tremor, rigidity, dystonia, dyarthria, and gait. The Total Score was obtained by summing these subscores. Eighty-six patients were included in our analysis, with a mean follow-up of 34.7 months. Retrospectively, the analysis compared scaled and unscaled sign subscores at enrollment and follow-up with change in the Total Score, using a generalized estimating equations approach. RESULTS: In the primary analysis, improvement in the Total Score was best predicted by sign subscores for tremor (beta -0.7, p = 0.006), gait abnormalities (beta -3.7, p < 0.001), and speech (beta = -1.3, p = 0.05). Dystonia (beta = 1.8, p < 0.001) and facial expression (beta = 1.9, p = 0.03) were associated with worsening Total Score. Of the motor signs followed individually, dystonia proved most resistant to treatment. CONCLUSIONS: This is the first large-scale prospectively acquired study assessing prognostic significance of specific neurologic signs in WD. Our data support the historical observations that tremor is a favorable prognostic sign while dystonia is relatively refractory to treatment in WD. PMID- 21641266 TI - Genetic characteristics of leucine-rich repeat kinase 2 (LRRK2) associated Parkinson's disease. AB - Parkinson's disease (PD) is a common neurodegenerative disorder characterized by the progressive and selective degeneration of nigrostriatal dopaminergic neurons. The discovery of at least six PD-causing genes in predominantly early-onset forms of the disorder has cemented a genetic component to the etiology. Notably, the discovery of mutations in the LRRK2 gene in patients presenting with typical 'sporadic' PD with ages at onset in their sixties and seventies has shifted paradigms in the field of PD research. The G2019S mutation in LRRK2 has been found in diverse populations worldwide and usually resides on a common haplotype revealing that many of these individuals share a common ancestor, probably of Middle Eastern origin. The only validated coding susceptibility alleles for PD, G2385R and R1628P, are both in this gene but to date have been found exclusively in Asian populations. Concomitant with genetic testing for PD is the need for appropriate and informed genetic counseling. Families of patients with LRRK2 mutations and susceptibility alleles need to be informed about the current lack of disease preventative strategies and the implications surrounding incomplete penetrance. In summary, single-handedly LRRK2 has had a major impact on the field of PD research and the findings have been of interest to both clinicians and scientists. We anticipate that other genes of such major impact exist for PD and look forward to their discovery. PMID- 21641267 TI - Long-term outcome of atlas-based lesion of posterior zona incerta in secondary hemidystonia. PMID- 21641268 TI - The relationship of transversus abdominis and lumbar multifidus clinical muscle tests in patients with chronic low back pain. AB - INTRODUCTION: Previous research of transversus abdominis (TrA) and multifidus muscle function in the presence of chronic low back pain (LBP) has investigated these muscles in isolation. In clinical practice, it is assumed that a relationship exists between these muscles and so they are often assessed and rehabilitated together. However, no studies have tested or documented this association. This study aimed to examine the relationships between clinical muscle testing and other measures taken in the course of a clinical assessment at a back clinic. METHODS: This retrospective chart audit examined the files of 82 patients (40 Males, 42 Females) for results of clinical tests of TrA and multifidus muscle contraction, multifidus muscle size measurements and other clinical measures such as distribution of pain and pain on manual examination. RESULTS: The ability to contract multifidus was related to the ability to contract TrA with the odds of a good contraction of multifidus being 4.5 times higher for patients who had a good contraction of TrA. A poor ability to contract multifidus was related to poor TrA contraction. Patients with unilateral LBP had more multifidus muscle asymmetry (11.6%) than those with bilateral/central pain (0.01%) and had a poor multifidus contraction on the affected side (p < 0.01). No other significant relationships were found. DISCUSSION & CONCLUSION: Current clinical practice of assessment and rehabilitation of both TrA and multifidus muscles in patients with chronic LBP is supported by the findings of this study. Future studies may investigate if a neurophysiological relationship exists between these muscles. PMID- 21641269 TI - Mediastinal atlas creation from 3-D chest computed tomography images: application to automated detection and station mapping of lymph nodes. AB - One important aspect of lung cancer staging is the assessment of mediastinal lymph nodes in 3-D chest computed tomography (CT) images. In the current clinical routine this is done manually by analyzing the 3-D CT image slice by slice to find nodes, evaluate them quantitatively, and assign labels to them for describing the clinical and pathologic extent of metastases. In this paper we present a method to automate the process of lymph node detection and labeling by creation of a mediastinal average image and a novel lymph node atlas containing probability maps for mediastinal, aortic, and N1 nodes. Utilizing a fast deformable registration approach to match the atlas with CT images of new patients, our method can maintain an acceptable runtime. In comparison to previously published methods for mediastinal lymph node detection and labeling it also shows a good sensitivity and positive predictive value. PMID- 21641270 TI - Context specific descriptors for tracking deforming tissue. AB - In minimally invasive surgery, deployment of motion compensation, dynamic active constraints and adaptive intra-operative guidance require accurate estimation of deforming tissue in 3D. To this end, the use of vision-based techniques is advantageous in that it does not require the integration of additional hardware to the existing surgical settings. Deformation can be recovered by tracking features on the surface of the tissue. Existing methods are mostly based on ad hoc machine vision techniques that have generally been developed for rigid scenes or use pre-determined models with parameters fine tuned to specific surgical settings. In this work, we propose a novel tracking technique based on a context specific feature descriptor. The descriptor can adapt to its surroundings and identify the most discriminate information for tracking. The feature descriptor is represented as a decision tree and the tracking process is formulated as a classification problem for which log likelihood ratios are used to improve classifier training. A recursive tracking algorithm obtains examples of tissue deformation used to train the classifier. Additional training data is generated by geometric and appearance modelling. Experimental results have shown that the proposed context specific descriptor is robust to drift, occlusion, and changes in orientation and scale. The performance of the algorithm is compared with existing tracking algorithms and validated with both simulated and in vivo datasets. PMID- 21641271 TI - Primary burning mouth syndrome in oral submucous fibrosis patients. PMID- 21641272 TI - Foxp3 expressed by tongue squamous cell carcinoma cells correlates with clinicopathologic features and overall survival in tongue squamous cell carcinoma patients. AB - The forkhead transcription factor, Foxp3, has been identified as a key player in CD4(+)CD25(+)Foxp3(+) regulatory T cells (Tregs) function and a definitive marker of Tregs. Recently, it was reported that Foxp3 could be expressed by tumor cells themselves. The present study was to investigate the expression of Foxp3 in tongue squamous cells carcinoma (TSCC) cells and its clinical significance. In this study, the expression of Foxp3 by TSCC cells was demonstrated in TSCC tissue samples and three TSCC cell lines using immunohistochemical staining, realtime PCR and Western blotting, and its clinical significance were statistically analyzed. The immunohistochemical assay in TSCC paraffin-embedded samples showed positive staining in 48 of 81 (59.3%) cases. The expression was significantly associated with pathologic differentiation (P=0.040) and T stage (P=0.000), and furthermore, inversely associate with patient survival (P=0.021). Multivariate analysis (Cox regression) suggested that Foxp3 expression in TSCC cells was an independent prognostic indicator for TSCC (P=0.032). PMID- 21641273 TI - EPR and optical studies of vanadium doped potassium dihydrogen citrate (C6H7KO7) single crystal. AB - An EPR and optical studies of VO2+ doped potassium dihydrogen citrate (PDHC) single crystals have been carried out at room temperature. It crystallizes in triclinic symmetry with the unit cell dimensions: a=11.343A, b=13.078A, c=6.272A, alpha=89.79 degrees , beta=94.36 degrees , gamma=104.2 degrees . The angular variation of EPR spectra have shown that two different VO2+ complexes are located in different chemical environments and each environment contains one magnetically VO2+ site occupying substitutional position in the lattice and show very high angular dependence. PMID- 21641274 TI - Viral factors influencing the outcome of human cytomegalovirus infection in liver transplant recipients. AB - BACKGROUND: Cytomegalovirus (CMV) remains the leading viral cause of disease following orthotopic liver transplantation (OLT) despite the availability of antiviral agents for prophylaxis and therapy. OBJECTIVE: Examine the viral factors that influence the outcome of CMV infection following valganciclovir prophylaxis or laboratory-guided preemptive therapy in OLT recipients. STUDY DESIGN: The value of valganciclovir prophylaxis and laboratory-guided preemptive therapy for the prevention of CMV infection and disease was observed in 64 OLT recipients. Prophylaxis was given to all CMV seronegative recipients receiving a liver from a seropositive donor (D+R-; n=15), and all other recipients were randomised to receive either prophylaxis (n=24) or laboratory-guided preemptive therapy (n=25). Recipients were monitored for CMV DNAemia, viral load, emergence of antiviral resistant strains and co-infections. RESULTS: CMV end-organ disease and antiviral resistant strains only occurred in D+R- recipients despite the use of prophylaxis in these patients. The D+R- recipients commencing prophylaxis immediately following transplantation had better outcomes compared to those for whom prophylaxis was delayed due to renal impairment. Prophylaxis reduced the incidence of CMV DNAemia, persistent infection, and high viral loads for CMV seropositive (D-R+and D+R+) recipients, but laboratory-guided preemptive therapy effectively controlled CMV infection and prevented disease in these OLT recipients. CONCLUSION: Delaying the commencement of valganciclovir prophylaxis may be associated with worse outcomes for high-risk OLT recipients. Laboratory guided pre-emptive therapy remains an alternative approach for seropositive recipients at lower risk of CMV disease. PMID- 21641275 TI - Polyarthritis in two travellers returning from Australia. PMID- 21641276 TI - Problems and challenges in the diagnosis of vertical infection with human cytomegalovirus (CMV): lessons from two accidental cases. AB - Human cytomegalovirus (CMV) is considered as the most common cause of congenital infection in humans and the overall burden for the public health system is rather high. About 1/10 of vertically infected newborns present or develop severe signs of cytomegalic inclusion disease (CID), with the classical triad of chorioretinitis, microcephaly and cerebral calcifications. However the most symptomatic cases are detected postnatal and methods of diagnostic virology raised the questions for the gold standard in laboratory screening. The current problems in diagnosis and therapy are outlined in two different cases: An acute primary CMV infection with no clinical signs of illness in both mother and child and a secondary CMV-infection resulting in necrotizing CMV encephalitis in the fetus. Beside virus detection in whole blood samples and other fluids, newly adopted laboratory assays like the destination of CMV-IgG avidity were necessary. Furthermore a serologic screening for pregnant women should be implicated routinely. Passive IgG treatment of the mother was helpful but the ultimate goal in prevention of congenital CMV infection is to develop a vaccine, which would be administered to seronegative women. PMID- 21641277 TI - In vitro and in vivo characterization of a new enterovirus type 71-specific human intravenous immunoglobulin manufactured from selected plasma donors. AB - BACKGROUND: Enterovirus type 71 (EV71) causes large outbreaks with significant mortality among young children, and no specific antiviral treatment is currently available. Antibody-based therapy represents a promising alternative strategy for lethal EV71 infection. Our previous data has shown that anti-EV71 neutralization antibodies were present in a significant proportion of blood donors in China. OBJECTIVES: To produce a new human intravenous immunoglobulin (IVIG) product containing high titer anti-EV71 neutralizing antibodies and investigate its therapeutic efficacy against lethal EV71 infection in a murine model. STUDY DESIGN: Plasma units that contained high titer neutralizing antibodies from selected Chinese donors were pooled and processed into pharmaceutical grade IVIG preparations according to the standard procedure. The efficacy of these EV71 specific IVIG product was characterized in vitro by neutralization assay and in vivo by suckling mouse protection testing. The therapeutic effects against lethal EV71 challenge were further assayed in a suckling mouse model. RESULTS: About 12% of the normal plasma units were selected and pooled to manufacture the EV71-IVIG preparations, and in vitro and in vivo efficacy data showed that these EV71 specific IVIG preparations were enriched with neutralizing antibodies against EV71. Furthermore, treatment with EV71-specific IVIG was evidenced to confer protection against lethal EV71 challenge in a dose- and time-dependent manner in the suckling mouse model. CONCLUSIONS: This preclinical study indicates that these "tailor-made" EV71-IVIG preparations manufactured from selected plasma donors in EV71-endemic areas may represent a promising therapeutic option for the lethal EV71 infections, and further clinical trials should be warranted in the future. PMID- 21641278 TI - Molecular biological researches of Kuro-Koji molds, their classification and safety. AB - To assess the position of Kuro-Koji molds in black Aspergillus, we performed sequence analysis of approximately 2500 nucleotides of partial gene fragments, such as histone 3, on a total of 57 Aspergillus strains, including Aspergillus kawachii NBRC 4308, 12 Kuro-Koji molds isolated from awamori breweries in Japan, Aspergillus niger ATCC 1015, and A. tubingensis ATCC10550. Sequence results showed that all black Aspergillus strains could be classified into 3 types, type N which includes A. niger ATCC 1015, type T which includes A. tubingensis ATCC 10550, and type L which includes A. kawachii NBRC 4308. Phylogenetic analysis showed these three types belong to different clusters. All 12 Kuro-Koji molds isolated from awamori breweries were classified as type L, thus we concluded type L represents the industrial Kuro-Koji molds. We found all type L strains lack the An15g07920 gene which is required for ochratoxin A biosynthesis in black Aspergillus. This sequence is present in the genome of A. niger CBS 513.88 and has homology to the polyketide synthase fragment of A. ochraceus which is involved in ochratoxin A biosynthesis. Based on the industrial importance and the safety of Kuro-Koji molds, we propose to classify the type L strains as Aspergillus luchuensis, as initially reported by Dr. Inui. PMID- 21641279 TI - Helicobacter pylori genotypes and types of gastritis in first-degree relatives of gastric cancer patients. AB - The frequency of Helicobacter pylori vacA alleles, cagA, and jhp0947 and their association with types and advanced forms of gastritis in 143 first-degree relatives of gastric cancer (GC) patients was assessed. The subjects included 64/143 with antral-predominant gastritis, 68/143 with pangastritis, and 11/143 with corpus-predominant gastritis, with or without atrophy or intestinal metaplasia (IM). Further classification included the severity of atrophy or IM. Group I (40/143) included the subjects with moderate-marked atrophy or IM, group II (58/143) those with no atrophy or IM, and group III (45/143) with mild atrophy or IM. The frequency of vacA s1 was 79.7%, vacA s2 20.3%, m1 49.7%, m2 50.3%, cagA 76.2%, and jhp0947 58%. The most prevalent combination was vacAs1 cagA (+) (65.7%) (P=0.001). Of the 143 subjects, 85 (59.4%) showed atrophy or IM, and 40/85 (47%) developed the moderate-marked atrophy or IM. No significant correlation was found between genotypes and the types of gastritis, non-atrophy, atrophy, or IM and severe forms of atrophy or IM (P>0.05). It is proposed that H. pylori genotype status might not be considered as an important determinant of the types and advanced forms of gastritis in the first-degree relatives of GC patients. PMID- 21641280 TI - Strategies for assessing and managing the adverse events of sorafenib and other targeted therapies in the treatment of renal cell and hepatocellular carcinoma: recommendations from a European nursing task group. AB - PURPOSE: As a group of European nurses familiar with treating patients with renal cell carcinoma (RCC) and hepatocellular carcinoma (HCC) using targeted/chemo- therapies, we aimed to review strategies for managing adverse events (AEs) associated with one targeted therapy, sorafenib. METHOD: Focusing on the AEs we considered the most difficult to manage (hand-foot skin reaction [HFSR], diarrhoea, fatigue and mucositis/stomatitis), we reviewed the literature to identify strategies relevant to sorafenib. Given the paucity of published work, this included strategies concerning targeted agents in general. This information was supplemented by considering the wider literature relating to management of these AEs in other tumour types and similar toxicities experienced during conventional anti-cancer therapy. Together with our own experience, this information was used to compile an AE management guide to assist nurses caring for patients receiving sorafenib. RESULTS: Our collated experience suggests the most commonly reported AEs with sorafenib and other targeted agents are HFSR, diarrhoea, fatigue, rash and mucositis/stomatitis; these generally have an acute (appearing at ~0-1 months) or delayed onset (appearing at ~3 months). Most management strategies in the literature were experience-based rather than arising from controlled studies. However, strategies based on controlled studies are available for HFSR and mucositis/stomatitis. CONCLUSIONS: Evidence, especially from controlled studies, is sparse concerning management of AEs associated with sorafenib and other targeted agents in RCC/HCC. However, recommendations can be made based on the literature and clinical experience that encompasses targeted and conventional therapies, particularly in the case of non-specific toxicities e.g. diarrhoea and fatigue. PMID- 21641282 TI - Prevalence of unruptured intracranial aneurysms, with emphasis on sex, age, comorbidity, country, and time period: a systematic review and meta-analysis. AB - BACKGROUND: Unruptured intracranial aneurysms (UIAs) are increasingly detected and are an important health-care burden. We aimed to assess the prevalence of UIAs according to family history, comorbidity, sex, age, country, and time period. METHODS: Through searches of PubMed, Embase, and Web of Science we updated our 1998 systematic review up to March, 2011. We calculated prevalences and prevalence ratios (PRs) with random-effects binomial meta-analysis. We assessed time trends with year of study as a continuous variable. FINDINGS: We included 68 studies, which reported on 83 study populations and 1450 UIAs in 94 912 patients from 21 countries. The overall prevalence was estimated as 3.2% (95% CI 1.9-5.2) in a population without comorbidity, with a mean age of 50 years, and consisting of 50% men. Compared with populations without the comorbidity, PRs were 6.9 (95% CI 3.5-14) for autosomal dominant polycystic kidney disease (ADPKD), 3.4 (1.9-5.9) for a positive family history of intracranial aneurysm of subarachnoid haemorrhage, 3.6 (0.4-30) for brain tumour, 2.0 (0.9-4.6) for pituitary adenoma, and 1.7 (0.9-3.0) for atherosclerosis. The PR for women compared with men was 1.61 (1.02-2.54), with a ratio of 2.2 (1.3-3.6) in study populations with a mean age of more than 50 years. Compared with patients older than 80 years, we found no differences by age, except for patients younger than 30 years (0.01, 0.00-0.12). Compared with the USA, PRs were similar for other countries, including Japan (0.8, 0.4-1.7) and Finland (1.0, 0.4-2.4). There was no statistically significant time trend. INTERPRETATION: The prevalence of UIAs is higher in patients with ADPKD or a positive family history of intracranial aneurysm of subarachnoid haemorrhage than in people without comorbidity. In Finland and Japan, the higher incidence of subarachnoid haemorrhage is not explained by a higher prevalence of UIAs, implicating higher risks of rupture. FUNDING: Julius Centre for Health Sciences and Primary Care and Department of Neurology and Neurosurgery, University Medical Centre, Utrecht. PMID- 21641281 TI - Meticillin-resistant Staphylococcus aureus with a novel mecA homologue in human and bovine populations in the UK and Denmark: a descriptive study. AB - BACKGROUND: Animals can act as a reservoir and source for the emergence of novel meticillin-resistant Staphylococcus aureus (MRSA) clones in human beings. Here, we report the discovery of a strain of S aureus (LGA251) isolated from bulk milk that was phenotypically resistant to meticillin but tested negative for the mecA gene and a preliminary investigation of the extent to which such strains are present in bovine and human populations. METHODS: Isolates of bovine MRSA were obtained from the Veterinary Laboratories Agency in the UK, and isolates of human MRSA were obtained from diagnostic or reference laboratories (two in the UK and one in Denmark). From these collections, we searched for mecA PCR-negative bovine and human S aureus isolates showing phenotypic meticillin resistance. We used whole-genome sequencing to establish the genetic basis for the observed antibiotic resistance. FINDINGS: A divergent mecA homologue (mecA(LGA251)) was discovered in the LGA251 genome located in a novel staphylococcal cassette chromosome mec element, designated type-XI SCCmec. The mecA(LGA251) was 70% identical to S aureus mecA homologues and was initially detected in 15 S aureus isolates from dairy cattle in England. These isolates were from three different multilocus sequence type lineages (CC130, CC705, and ST425); spa type t843 (associated with CC130) was identified in 60% of bovine isolates. When human mecA negative MRSA isolates were tested, the mecA(LGA251) homologue was identified in 12 of 16 isolates from Scotland, 15 of 26 from England, and 24 of 32 from Denmark. As in cows, t843 was the most common spa type detected in human beings. INTERPRETATION: Although routine culture and antimicrobial susceptibility testing will identify S aureus isolates with this novel mecA homologue as meticillin resistant, present confirmatory methods will not identify them as MRSA. New diagnostic guidelines for the detection of MRSA should consider the inclusion of tests for mecA(LGA251). FUNDING: Department for Environment, Food and Rural Affairs, Higher Education Funding Council for England, Isaac Newton Trust (University of Cambridge), and the Wellcome Trust. PMID- 21641283 TI - Prevalence of and risk factors for intracranial aneurysms. PMID- 21641284 TI - Comparing population distributions from bin-aggregated sample data: An application to historical height data from France. AB - We develop a methodology to estimate underlying (continuous) population distributions from bin-aggregated sample data through the estimation of the parameters of mixtures of distributions that allow for maximal parametric flexibility. The statistical approach we develop enables comparisons of the full distributions of height data from potential army conscripts across France's 88 departments for most of the nineteenth century. These comparisons are made by testing for differences-of-means stochastic dominance. Corrections for possible measurement errors are also devised by taking advantage of the richness of the data sets. Our methodology is of interest to researchers working on bin aggregated or histogram-type data, something that is still widely done since much of the information that is publicly available is in that form, often due to restrictions based on confidentiality concerns. PMID- 21641285 TI - Social inequality and the biological standard of living: an anthropometric analysis of Swiss conscription data, 1875-1950. AB - We analyze the first representative series of individual measurements of the height of Swiss conscripts for the years 1875-1950. We find that average height followed a general upward time trend, but the economic downturn in the 1880s slowed down the increase in rural average-heights while the economic crisis subsequent to World War I had only a minor effect. Moreover, social-class affiliation was the most important determinant of differences in the biological standard of living, with class and regional disparities remaining constant, for the most part, during the observation period. Lower-class individuals' ability to overcome economic stress was limited, with the result that their biological standard of living, as reflected in the cyclicality of deviations from average height, was likely to be affected by cycles in economic activity. PMID- 21641286 TI - [Therapeutic approach to dyslipidemia and goal achievement in a Spanish population with type 2 diabetes without cardiovascular disease]. AB - OBJECTIVE: To assess the therapeutic approach and lipid goal achievement in a spanish diabetic population at high cardiovascular risk. SUBJECTS AND METHODS: A multicenter, descriptive, cross-sectional study consecutively recruited the first 10 patients who attended the primary care office and had been seen in the 12 months prior to the study visit. Inclusion criteria were type 2 diabetes without cardiovascular disease, LDL cholesterol levels <=160mg/dL, triglyceride levels <=600mg/dL, and at least one of the following: retinopathy, albuminuria, current smoking, or hypertension. RESULTS: A total of 2412 patients were evaluated (aged 61.3+/-8.3 years, 46.8% women, diabetes duration 8.6+/-7.4 years). As compared to the previous visit (8.1+/-5 months before), the proportion of patients who achieved LDL-C levels <100mg/dL (22.7% vs 28.6%), non-HDL-C levels <130mg/dL (27.7% vs 33.8%) and both goals (17.6% vs 22.1%) significantly increased at the time of assessment. Statins were the most widely prescribed lipid-lowering drugs (65.5%) and the lipid-lowering drug was changed from the previous visit in 38.7% of patients, drug dosage was increased in 17.3%, and another drug was added in 5%. CONCLUSION: Use of more potent statins and higher statin doses were the most commonly used therapeutic strategies for improving control of dyslipidemia in patients with type 2 diabetes, but these changes were clearly inadequate to achieve lipid goals in most patients with type 2 diabetes. PMID- 21641287 TI - [Direct health care costs in patients with type 2 diabetes mellitus six months after starting insulin treatment in Spain: the INSTIGATE study]. AB - BACKGROUND AND OBJECTIVES: The INSTIGATE study was designed to assess direct health care costs incurred by patients with type 2 diabetes mellitus (T2DM) who start insulin therapy in Spain. It was a multicenter, observational, non interventional, prospective study. METHODS: Direct costs per patient in standard clinical practice were assessed for 6 months before and after the start of insulin therapy from the perspective of the Spanish health care system perspective. A total of 188 patients (42.6% women) with a mean age of 65.3 years, a mean body mass index of 29.7 kg/m(2), and a mean disease duration of 10.7 years were assessed. RESULTS: Before insulin therapy was started, mean (standard deviation) values of various clinical parameters were: hemoglobin A(1c) (%), 9.22 (1.58); fasting plasma glucose (mmol/L), 12.03 (3.62); and total cholesterol (mmol/L), 4.90 (1.1). These values decreased after insulin therapy was started. Mean total direct health care costs per patient 6 months before and after insulin start were ?639 and ?1,110, respectively. Mean total costs 6 months after insulin was started included costs of hospitalization (30.5%, ?339), insulin (16.2%, ?180), primary care (14.3%, ?159), blood glucose monitoring (13.8%, ?153), specialized care (13.3%, ?148), oral antidiabetics (7.8%, ?87), and other diabetes-related treatments (3.9%, ?43). CONCLUSIONS: The clinical outcomes of T2DM patients improved after insulin therapy was started. This improvement was associated to increases in resource utilization and direct health care costs in the first 6 months of insulin therapy. PMID- 21641288 TI - [TRAMOMTANA (Multidisciplinary treatment of morbid obesity: medication, behavioral therapy, nutritional support, and physical activity). From question to reality in an investigator-initiated clinical trial (II)]. AB - Implementation of an intensive, multidisciplinary weight loss program in patients with morbid obesity is reported. This program is based on behavioral changes, lifestyle intervention, medication, and group therapy sessions. Our objective is to show that the results achieved with this two-year weight loss program will be at least similar to those achieved with bariatric surgery in patients with morbid obesity. We also intend to show that this multidisciplinary treatment induces an improvement in the comorbidity rate associated to smaller costs for our national health system. PMID- 21641289 TI - Myth: tocolysis for prevention of preterm birth has a major role in modern obstetrics. AB - Tocolytics are widely used to reduce uterine activity in the context of preterm labour. Growing evidence that bacterial colonization of fetal membranes and amniotic fluid triggers an inflammatory response in mother and fetus and leads to preterm labour and long term neurological and respiratory complications in the neonate also raises questions about the desirability of prolonging pregnancy in this context. Combined with recent meta-analyses that fail to demonstrate improvements in neonatal outcome with tocolytic use, and a poor maternal/fetal side-effect profile, the case for continued use of these drugs needs to be questioned. PMID- 21641290 TI - Echocardiographic assessment of interventricular and intraventricular mechanical synchrony in normal dogs. AB - OBJECTIVES: The aims of this prospective study are to (1) generate normal ranges for interventricular and intraventricular mechanical synchrony in dogs, and (2) generate normal ranges for tissue Doppler imaging (TDI) velocity imaging and speckle tracking strain imaging assessment of segmental intraventricular mechanical synchrony in dogs. ANIMALS: 10 prospectively recruited healthy dogs. METHODS: Dogs were excluded if they had abnormal historical, physical examination, echocardiographic, ECG or systolic blood pressure findings. Interventricular mechanical synchrony was assessed using time difference between left and right ventricular pre-ejection periods. Intraventricular mechanical synchrony was assessed using both M-mode and color M-mode septal to posterior wall mechanical delay (SPWMD). Intraventricular segmental mechanical synchrony was assessed using both color TDI and speckle tracking strain analysis of segmental myocardial motion during systole and diastole. RESULTS: All synchrony measures were found to be independent of age or body weight. Normal range for mechanical interventricular synchrony was found to be -10.2 to 12.6 ms. Assessment of mechanical intraventricular synchrony using either M-mode or color M-mode SPWMD was found to be associated with an extremely wide normal range, limiting clinical applicability. Normal ranges for segmental intraventricular mechanical synchrony assessed using either color TDI or speckle tracking were found to be comparable to those published for human subjects. CONCLUSIONS: Interventricular and intraventricular mechanical synchrony in dogs is independent of age and body weight. The normal ranges identified in this study form a basis for assessment of normal versus abnormal mechanical synchrony in canine cardiovascular disease patients. PMID- 21641291 TI - Allele frequencies of 15 STRs in the Calchaqui Valleys population (North-Western Argentina). AB - Allele frequencies for 15 short tandem repeat (STR) loci were obtained from a sample of 110 individuals from the Calchaqui Valleys population (North-Western Argentina). The combined power of exclusion and combined power of discriminating for the 15 tested STR loci were 0.999964 and 0.9999999999999998, respectively. Matching probability was 1 in 4.58 * 10(15). Therefore, it may be concluded that the set of 15 STRs included in the AmpF STR Identifiler kit, represents a powerful tool for forensic applications, paternity testing and population genetics studies in the Calchaqui Valleys population. PMID- 21641292 TI - Open grade III fractures of femoral shaft: Outcome after early reamed intramedullary nailing. AB - BACKGROUND: Open grade III fractures of femur are a challenging therapeutic problem as most of them are associated with multiple trauma. Method used for skeletal stabilization of these fractures should limit further soft tissue damage, bacterial spread and morbidity in addition to its ease of application, providing mechanical support and restoring normal alignment. METHODS: Forty-six patients with open grade III fractures of femoral shaft were included in the study. There were 10 grade IIIA, 34 grade IIIB and two grade IIIC fractures that were treated with early reamed intramedullary nailing with adequate management of soft tissue. Patients were reviewed retrospectively. RESULTS: Mean time of union was 27 weeks. Infection rate was 4% and non-union rate was 9%. There were two cases (4%) with limb shortening. No case of angular or rotational deformity was noted. CONCLUSION: Early reamed intramedullary nailing for open grade III fractures of femoral shaft, after giving due respect to the soft tissue injury, gives satisfactory results with proper alignment, good range of motion, short rehabilitation period and low infection rate. LEVEL OF EVIDENCE: Level IV. Retrospective therapeutic study. PMID- 21641293 TI - Larger than life: overestimation of object size is moderated by personal relevance in obsessive-compulsive disorder. AB - BACKGROUND AND OBJECTIVES: Along with other cognitive biases overestimation of threat (OET) has been implicated in the pathogenesis of obsessive-compulsive disorder (OCD). The present study investigated whether OET would not only manifest in cognitive distortions but, also in overestimations of the object size of disorder-related visual objects. METHODS: A total of 65 participants with OCD and 55 healthy controls who were recruited via OCD online forums underwent an incidental learning paradigm consisting of two blocks. In Block 1, participants were asked to rate the valence and the personal relevance for individual OCD concerns of 40 pictures which varied in size. Differences in size, however, were not explicitly communicated to the participants. Stimuli were selected from four categories: 1. neutral, 2. fear-related but OCD-unrelated, 3. washing (OCD related), and 4. checking (OCD-related). In Block 2, participants were asked to recollect the original size of each stimulus (depicted as a small thumbnail) on a seven point scale. RESULTS: Whereas few group differences emerged for pre-defined OCD items, OCD-relevant items (individual judgments) were judged as significantly larger by patients with OCD relative to controls. The opposite pattern emerged for neutral items. LIMITATIONS: The sample was recruited via online forums and had probable but not externally validated diagnoses of OCD. No psychiatric control group was recruited. CONCLUSIONS: The present study indicates that OET may extend to neuropsychological tasks. Further research is needed to pinpoint whether OET occurs at the level of encoding suggesting a perceptual bias and/or occurs at the level of retrieval suggesting a memory bias. PMID- 21641294 TI - Influence of triphenyltin exposure on the hypothalamus-pituitary-gonad axis in male Sebastiscus marmoratus. AB - Both triphenyltin (TPT) and tributyltin (TBT) have been used as ingredients of antifouling biocides. However, far fewer studies addressing the reproductive toxicity of TPT on fishes are available than for TBT. The present study was conducted to investigate the effects of TPT at environmentally relevant concentrations on testicular development in male rockfish Sebastiscus marmoratus and to gain insight into its mechanism of action. After exposure for 48 days, the gonadosomatic index had decreased, and there was a reduced number of mature sperm and an abundance of the late stages of spermatocysts in the testes. Although the testosterone levels in the testes were elevated and the 17beta-estradiol levels were decreased, spermatogenesis was suppressed. The activity of gamma-glutamyl transpeptidase (which is used as a Sertoli cell marker) was decreased after TPT exposure, and serious interstitial fibrosis was observed in the interlobular septa of the testes exposed to TPT. The increased expression of cGnRH-II (chicken II type gonadotropin-releasing hormone) and sGnRH (salmon-type GnRH), and the decreased expression of LHbeta (luteinizing hormone) in the fish brains were detected. The expression of FSHbeta (follicle-stimulating hormone) was decreased at day 21, while was increased slightly at day 48. The changes of cGnRH-II, sGnRH, FSHbeta and LHbeta mRNA levels might have mainly resulted from the alteration of the sex steroids via feedback mechanisms. The decrease of the FSHbeta mRNA might have been one of the reasons causing the dysfunction of Sertoli cells, which play a critical role during spermatogenesis. The results suggested that TPT could perturb the function of hypothalamus-pituitary-gonad axis, and inhibiting the spermatogenesis. PMID- 21641295 TI - Differentially expressed proteins in zebrafish liver cells exposed to copper. AB - Copper is an essential element for normal cellular processes in most eukaryotic organisms, but is toxic in excessive amounts. Different organisms vary in their ability to tolerate copper ions. We have previously studied the mechanism of copper toxicity to a copper tolerance cell line, Hepa T1, from tilapia using a proteomic approach. To compare the differences of proteins' regulation between copper tolerant and sensitive species after copper treatment, the zebrafish liver cell line (ZFL) was used as a model in this study to investigate the mechanism of copper toxicity to zebrafish. After conducting similar experimental procedures in previous Hepa T1 studies, 72 different proteins were identified to be regulated by Cu(2+) (100 MUM and 200 MUM). More than 50% of these proteins were also found with differentially expressed Hepa T1, indicating that the toxicity mechanism between zebrafish and tilapia was partially conserved. However, the regulation of several proteins in ZFL, related to the reactive oxygen species (ROS) effect, mitochondrion copper transportation and stress response, was quite different from that in tilapia. PMID- 21641296 TI - Intersex and reproductive impairment of wild fish exposed to multiple municipal wastewater discharges. AB - The Grand River watershed in Ontario, Canada, receives and assimilates the outflow of 29 Municipal Wastewater Effluent (MWWE) discharges which is a mixture of domestic and industrial wastes. The purpose of this study was to investigate the cumulative impact of multiple sewage discharges on populations of wild fish. In field studies, responses of fish populations and individual fish responses in terms of growth (condition factor), reproduction (in vitro sex steroid production, gonadosomatic indices, histology [cellular development and intersex]) were assessed upstream and downstream of two municipal discharges. Fish [Greenside Darters Etheostoma blennioides and Rainbow Darters E. caeruleum] collected downstream of two municipal wastewater plants had the potential to have greater growth (longer and heavier) when compared to reference fish collections regardless of sex. Fish were not assimilating additional anthropogenic resources into energy storage (increased condition, liver somatic index). Impacts on ovarian development appeared to be minor with no differences in growth, steroid production or cellular development. Sewage exposed male fish were experiencing impairment in the capacity to produce testosterone and 11-ketotestosterone in vitro, and in cellular development (GSI, intersex). Male darters of both species collected in the upstream agricultural region demonstrated no evidence of intersex whereas our urban reference sites had incidence of intersex of up to 20%. Rates of intersex were elevated downstream of both sewage discharges studied (33% and>60%, respectively). Lower rates of intersex at the intermediate sites, and then increases downstream of second sewage discharge suggests that fish populations have to potential to recover prior to exposure to the second sewage effluent. Pre-spawning darters demonstrated dramatically higher incidence of intersex in the spring at both urban reference sites (33% and 50%, respectively), and increased more so downstream of the near-field and far-field exposure sites (60% and 100%, respectively). These findings suggest that the compounds released in STP effluents have a tendency to act on the male reproductive system. These effects may become more pronounced as projected human population growth will require the aquatic environment to assimilate an increasing amount of sewage waste. PMID- 21641297 TI - Viral capsid equilibrium dynamics reveals nonuniform elastic properties. AB - The long wavelength, low-frequency modes of motion are the relevant motions for understanding the continuum mechanical properties of biomolecules. By examining these low-frequency modes, in the context of a spherical harmonic basis set, we identify four elastic moduli that are required to describe the two-dimensional elastic behavior of capsids. This is in contrast to previous modeling and theoretical studies on elastic shells, which use only the two-dimensional Young's modulus (Y) and the bending modulus (kappa) to describe the system. Presumably, the heterogeneity of the structure and the anisotropy of the biomolecular interactions lead to a deviation from the homogeneous, isotropic, linear elastic shell theory. We assign functional relevance of the various moduli governing different deformation modes, including a mode primarily sensed in atomic force microscopy nanoindentation experiments. We have performed our analysis on the T = 3 cowpea chlorotic mottle virus and our estimate for the nanoindentation modulus is in accord with experimental measurements. PMID- 21641298 TI - A model for segregation of chromatin after replication: segregation of identical flexible chains in solution. AB - We study the segregation of two long chains from parallel but randomly twisted start conformations under good solvent conditions using Monte Carlo simulations to mimic chromatin segregation after replication in eukaryotic cells in the end of prophase. To measure the segregation process, we consider the center-of-mass separation between the two chains and the average square distance between the monomers which were connected before segregation starts. We argue that segregation is dominated by free diffusion of the chains, assuming that untwisting can be achieved by Rouse-like fluctuations on the length scale of a twisted loop. Using scaling analysis, we find that chain dynamics is in very good agreement with the free diffusion hypothesis, and segregation dynamics follows this scaling nearly. Long chains, however, show retardation effects that can be described by a new (to us) dynamical exponent, which is slightly larger than the dynamical exponent for Rouse-like diffusion. Our results indicate that nearly free diffusion of chains during a timescale of a few Rouse-times can lead to segregation of chains. A main obstacle during segregation by free diffusion is random twists between daughter strands. We have calculated the number of twists formed by the daughter strands in the start conformations, which turns out to be rather low and increases only with the square-root of the chain length. PMID- 21641299 TI - Theoretical and computational investigation of flagellin translocation and bacterial flagellum growth. AB - The bacterial flagellum is a self-assembling filament, which bacteria use for swimming. It is built from tens of thousands of flagellin monomers in a self assembly process that involves translocation of the monomers through the flagellar interior, a channel, to the growing tip. Flagellum monomers are pumped into the filament at the base, move unfolded along the channel and then bind to the tip of the filament, thereby extending the growing flagellum. The flagellin translocation process, due to the flagellum maximum length of 20 MUm, is an extreme example of protein transport through channels. Here, we derive a model for flagellin transport through the long confining channel, testing the key assumptions of the model through molecular dynamics simulations that also furnish system parameters needed for quantitative description. Together, mathematical model and molecular dynamics simulations explain why the growth rate of flagellar filaments decays exponentially with filament length and why flagellum growth ceases at a certain maximum length. PMID- 21641300 TI - Robust entrainment of circadian oscillators requires specific phase response curves. AB - The circadian clocks keeping time in many living organisms rely on self-sustained biochemical oscillations entrained by external cues, such as light, to the 24-h cycle induced by Earth's rotation. However, environmental cues are unreliable due to the variability of habitats, weather conditions, or cue-sensing mechanisms among individuals. A tempting hypothesis is that circadian clocks have evolved so as to be robust to fluctuations in the signal that entrains them. To support this hypothesis, we analyze the synchronization behavior of weakly and periodically forced oscillators in terms of their phase response curve (PRC), which measures phase changes induced by a perturbation applied at different times of the cycle. We establish a general relationship between the robustness of key entrainment properties, such as stability and oscillator phase, on the one hand, and the shape of the PRC as characterized by a specific curvature or the existence of a dead zone, on the other hand. The criteria obtained are applied to computational models of circadian clocks and account for the disparate robustness properties of various forcing schemes. Finally, the analysis of PRCs measured experimentally in several organisms strongly suggests a case of convergent evolution toward an optimal strategy for maintaining a clock that is accurate and robust to environmental fluctuations. PMID- 21641301 TI - Orientation and polarity in collectively migrating cell structures: statics and dynamics. AB - Collective cell migration is often characterized by the spontaneous onset of multicellular protrusions (known as fingers) led by a single leader cell. Working with epithelial Madin-Darby canine kidney monolayers we show that cells within the fingers, as compared with the epithelium, are well oriented and polarized along the main finger direction, which suggests that these cells actively migrate. The cell orientation and polarity decrease continuously from the tip toward the epithelium over a penetration distance of typically two finger lengths. Furthermore, laser photoablation experiments at various locations along these fingers demonstrate that the cells in the fingers are submitted to a tensile stress whose value is larger close to the tip. From a dynamical point of view, cells entering a finger gradually polarize on timescales that depend upon their particular initial position. Selective laser nanosurgery of the leader lamellipodium shows not only that these structures need a leader to progress, but that this leader itself is the consequence of a prior self-organization of the cells forming the finger. These results highlight the complex interplay between the collective orientation within the fingers and the mechanical action of the leader. PMID- 21641302 TI - Coupling active hair bundle mechanics, fast adaptation, and somatic motility in a cochlear model. AB - One of the central questions in the biophysics of the mammalian cochlea is determining the contributions of the two active processes, prestin-based somatic motility and hair bundle (HB) motility, to cochlear amplification. HB force generation is linked to fast adaptation of the transduction current via a calcium dependent process and somatic force generation is driven by the depolarization caused by the transduction current. In this article, we construct a global mechanical-electrical-acoustical mathematical model of the cochlea based on a three-dimensional fluid representation. The global cochlear model is coupled to linearizations of nonlinear somatic motility and HB activity as well as to the micromechanics of the passive structural and electrical elements of the cochlea. We find that the active HB force alone is not sufficient to power high frequency cochlear amplification. However, somatic motility can overcome resistor-capacitor filtering by the basolateral membrane and deliver sufficient mechanical energy for amplification at basal locations. The results suggest a new theory for high frequency active cochlear mechanics, in which fast adaptation controls the transduction channel sensitivity and thereby the magnitude of the energy delivered by somatic motility. PMID- 21641303 TI - The endocochlear potential alters cochlear micromechanics. AB - Acoustic stimulation gates mechanically sensitive ion channels in cochlear sensory hair cells. Even in the absence of sound, a fraction of these channels remains open, forming a conductance between hair cells and the adjacent fluid space, scala media. Restoring the lost endogenous polarization of scala media in an in vitro preparation of the whole cochlea depolarizes the hair cell soma. Using both digital laser interferometry and time-resolved confocal imaging, we show that this causes a structural refinement within the organ of Corti that is dependent on the somatic electromotility of the outer hair cells (OHCs). Specifically, the inner part of the reticular lamina up to the second row of OHCs is pulled toward the basilar membrane, whereas the outer part (third row of OHCs and the Hensen's cells) unexpectedly moves in the opposite direction. A similar differentiated response pattern is observed for sound-evoked vibrations: restoration of the endogenous polarization decreases vibrations of the inner part of the reticular lamina and results in up to a 10-fold increase of vibrations of the outer part. We conclude that the endogenous polarization of scala media affects the function of the hearing organ by altering its geometry, mechanical and electrical properties. PMID- 21641304 TI - Cooperativity between integrin activation and mechanical stress leads to integrin clustering. AB - Integrins are transmembrane receptors involved in crucial cellular biological functions such as migration, adhesion, and spreading. Upon the modulation of integrin affinity toward their extracellular ligands by cytoplasmic proteins (inside-out signaling) these receptors bind to their ligands and cluster into nascent adhesions. This clustering results in the increase in the mechanical linkage among the cell and substratum, cytoskeleton rearrangements, and further outside-in signaling. Based on experimental observations of the distribution of focal adhesions in cells attached to micropatterned surfaces, we introduce a physical model relying on experimental numerical constants determined in the literature. In this model, allosteric integrin activation works in synergy with the stress build by adhesion and the membrane rigidity to allow the clustering to nascent adhesions independently of actin but dependent on the integrin diffusion onto adhesive surfaces. The initial clustering could provide a template to the mature adhesive structures. Predictions of our model for the organization of focal adhesions are discussed in comparison with experiments using adhesive protein microarrays. PMID- 21641305 TI - Entropy-based mechanism of ribosome-nucleoid segregation in E. coli cells. AB - In Escherichia coli, ribosomes concentrate near the cylindrical wall and at the endcaps, whereas the chromosomal DNA segregates in the more centrally located nucleoid. A simple statistical model recovers the observed ribosome-nucleoid segregation remarkably well. Plectonemic DNA is represented as a hyperbranched hard-sphere polymer, and multiple ribosomes that simultaneously translate the same mRNA strand (polysomes) are represented as freely jointed chains of hard spheres. There are no attractive interactions between particles, only excluded volume effects. At realistic DNA and ribosome concentrations, segregation arises primarily from two effects: the DNA polymer avoids walls to maximize conformational entropy, and the polysomes occupy the empty space near the walls to maximize translational entropy. In this complex system, maximizing total entropy results in spatial organization of the components. Due to coupling of mRNA to DNA through RNA polymerase, the same entropic effects should favor the placement of highly expressed genes at the interface between the nucleoid and the ribosome-rich periphery. Such a placement would enable efficient cotranscriptional translation and facile transertion of membrane proteins into the cytoplasmic membrane. Finally, in the model, monofunctional DNA polymer beads representing the tips of plectonemes preferentially locate near the cylindrical wall. This suggests that initiation of transcription may occur preferentially near the ribosome-rich periphery. PMID- 21641306 TI - Membrane thickness sensitivity of prestin orthologs: the evolution of a piezoelectric protein. AB - How proteins evolve new functionality is an important question in biology; prestin (SLC26A5) is a case in point. Prestin drives outer hair cell somatic motility and amplifies mechanical vibrations in the mammalian cochlea. The motility of mammalian prestin is analogous to piezoelectricity, in which charge transfer is coupled to changes in membrane area occupied by the protein. Intriguingly, nonmammalian prestin orthologs function as anion exchangers but are apparently nonmotile. We previously found that mammalian prestin is sensitive to membrane thickness, suggesting that prestin's extended conformation has a thinner hydrophobic height in the lipid bilayer. Because prestin-based motility is a mammalian specialization, we initially hypothesized that nonmotile prestin orthologs, while functioning as anion transporters, should be much less sensitive to membrane thickness. We found the exact opposite to be true. Chicken prestin was the most sensitive to thickness changes, displaying the largest shift in voltage dependence. Platypus prestin displayed an intermediate response to membrane thickness and gerbil prestin was the least sensitive. To explain these observations, we present a theory where force production, rather than displacement, was selected for the evolution of prestin as a piezoelectric membrane motor. PMID- 21641307 TI - The discovery of slowness: low-capacity transport and slow anion channel gating by the glutamate transporter EAAT5. AB - Excitatory amino acid transporters (EAATs) control the glutamate concentration in the synaptic cleft by glial and neuronal glutamate uptake. Uphill glutamate transport is achieved by the co-/countertransport of Na(+) and other ions down their concentration gradients. Glutamate transporters also display an anion conductance that is activated by the binding of Na(+) and glutamate but is not thermodynamically coupled to the transport process. Of the five known glutamate transporter subtypes, the retina-specific subtype EAAT5 has the largest conductance relative to glutamate uptake activity. Our results suggest that EAAT5 behaves as a slow-gated anion channel with little glutamate transport activity. At steady state, EAAT5 was activated by glutamate, with a K(m)= 61 +/- 11 MUM. Binding of Na(+) to the empty transporter is associated with a K(m) = 229 +/- 37 mM, and binding to the glutamate-bound form is associated with a K(m) = 76 +/- 40 mM. Using laser-pulse photolysis of caged glutamate, we determined the pre-steady state kinetics of the glutamate-induced anion current of EAAT5. This was characterized by two exponential components with time constants of 30 +/- 1 ms and 200 +/- 15 ms, which is an order of magnitude slower than those observed in other glutamate transporters. A voltage-jump analysis of the anion currents indicates that the slow activation behavior is caused by two slow, rate-limiting steps in the transport cycle, Na(+) binding to the empty transporter, and translocation of the fully loaded transporter. We propose a kinetic transport scheme that includes these two slow steps and can account for the experimentally observed data. Overall, our results suggest that EAAT5 may not act as a classical high-capacity glutamate transporter in the retina; rather, it may function as a slow-gated glutamate receptor and/or glutamate buffering system. PMID- 21641308 TI - Sterols have higher affinity for sphingomyelin than for phosphatidylcholine bilayers even at equal acyl-chain order. AB - The interaction between cholesterol and phospholipids in bilayer membranes is important for the formation and maintenance of membrane structure and function. However, cholesterol does not interact favorably with all types of phospholipids and, for example, prefers more ordered sphingomyelins (SMs) over phosphatidylcholines (PCs). The reason for this preference is not clear. Here we have studied whether acyl-chain order could be responsible for the preferred sterol interaction with SMs. Acyl-chain order was deduced from diphenylhexatriene anisotropy and from the deuterium order parameter obtained by (2)H-NMR on bilayers made from either 14:0/14:0((d27))-PC, or 14:0((d27))-SM. Sterol/phospholipid interaction was determined from sterol bilayer partitioning. Cholestatrienol (CTL) was used as a fluorescence probe for cholesterol, because its relative membrane partitioning is similar to cholesterol. When CTL was allowed to reach equilibrium partitioning between cyclodextrins and unilamellar vesicles made from either 14:0/14:0-PC or 14:0-SM, the molar-fraction partitioning coefficient (K(x)) was approximately twofold higher for SM bilayers than for PC bilayers. This was even the case when the temperature in the SM samples was raised to achieve equal acyl-chain order, as determined from 1,6 diphenyl-1,3,5-hexatriene (DPH) anisotropy and the deuterium order parameter. Although the K(x) did increase with acyl-chain order, the higher K(x) for SM bilayers was always evident. At equal acyl-chain order parameter (DPH anisotropy), the K(x) was also higher for 14:0-SM bilayers than for bilayers made from either 14:0/15:0-PC or 15:0-/14:0-PC, suggesting that minor differences in chain length or molecular asymmetry are not responsible for the difference in K(x). We conclude that acyl-chain order affects the bilayer affinity of CTL (and thus cholesterol), but that it is not the cause for the preferred affinity of sterols for SMs over matched PCs. Instead, it is likely that the interfacial properties of SMs influence and stabilize interactions with sterols in bilayer membranes. PMID- 21641309 TI - Minimal encounter time and separation determine ligand-receptor binding in cell adhesion. AB - The binding properties of biomolecules play a crucial role in many biological phenomena, especially cell adhesion. Whereas the attachment kinetics of soluble proteins is considered well known, complex behavior arises when protein molecules are bound to the cell membrane. We probe the hidden kinetics of ligand-receptor bond formation using single-molecule flow chamber assays and Brownian dynamics simulations. We show that, consistent with our recently proposed hypothesis, association requires a minimum duration of contact between the reactive species. In our experiments, ICAM-1 anchored on a flat substrate binds to anti-ICAM-1 coated onto flowing microbeads. The interaction potential between bead and substrate is measured by microinterferometry and is used as an ingredient to simulate bead movement. Our simulation calculates the duration of ligand-receptor contacts imposed by the bead movement. We quantitatively predict the reduction of adhesion probability measured for shorter tether length of the ligand or if a repulsive hyaluronan layer is added onto the surface. To account for our results, we propose that bond formation may occur in our system by crossing of a diffusive plateau in the energy landscape, on the timescale of 5 ms and an energy barrier of 5 k(B)T, before reaching the first detectable bound state. Our results show how to relate cell-scale behavior to the combined information of molecular reactivity and biomolecule submicron-scale environment. PMID- 21641310 TI - Biophysical mechanisms of endotoxin neutralization by cationic amphiphilic peptides. AB - Bacterial endotoxins (lipopolysaccharides (LPS)) are strong elicitors of the human immune system by interacting with serum and membrane proteins such as lipopolysaccharide-binding protein (LBP) and CD14 with high specificity. At LPS concentrations as low as 0.3 ng/ml, such interactions may lead to severe pathophysiological effects, including sepsis and septic shock. One approach to inhibit an uncontrolled inflammatory reaction is the use of appropriate polycationic and amphiphilic antimicrobial peptides, here called synthetic anti LPS peptides (SALPs). We designed various SALP structures and investigated their ability to inhibit LPS-induced cytokine secretion in vitro, their protective effect in a mouse model of sepsis, and their cytotoxicity in physiological human cells. Using a variety of biophysical techniques, we investigated selected SALPs with considerable differences in their biological responses to characterize and understand the mechanism of LPS inactivation by SALPs. Our investigations show that neutralization of LPS by peptides is associated with a fluidization of the LPS acyl chains, a strong exothermic Coulomb interaction between the two compounds, and a drastic change of the LPS aggregate type from cubic into multilamellar, with an increase in the aggregate sizes, inhibiting the binding of LBP and other mammalian proteins to the endotoxin. At the same time, peptide binding to phospholipids of human origin (e.g., phosphatidylcholine) does not cause essential structural changes, such as changes in membrane fluidity and bilayer structure. The absence of cytotoxicity is explained by the high specificity of the interaction of the peptides with LPS. PMID- 21641311 TI - DC-SIGN and influenza hemagglutinin dynamics in plasma membrane microdomains are markedly different. AB - DC-SIGN, a Ca(2+)-dependent transmembrane lectin, is found assembled in microdomains on the plasma membranes of dendritic cells. These microdomains bind a large variety of pathogens and facilitate their uptake for subsequent antigen presentation. In this study, DC-SIGN dynamics in microdomains were explored with several fluorescence microscopy methods and compared with dynamics for influenza hemagglutinin (HA), which is also found in plasma membrane microdomains. Fluorescence imaging indicated that DC-SIGN microdomains may contain other C-type lectins and that the DC-SIGN cytoplasmic region is not required for microdomain formation. Fluorescence recovery after photobleaching measurements showed that neither full-length nor cytoplasmically truncated DC-SIGN in microdomains appreciably exchanged with like molecules in other microdomains and the membrane surround, whereas HA in microdomains exchanged almost completely. Line-scan fluorescence correlation spectroscopy indicated an essentially undetectable lateral mobility for DC-SIGN but an appreciable mobility for HA within their respective domains. Single-particle tracking with defined-valency quantum dots confirmed that HA has significant mobility within microdomains, whereas DC-SIGN does not. By contrast, fluorescence recovery after photobleaching indicated that inner leaflet lipids are able to move through DC-SIGN microdomains. The surprising stability of DC-SIGN microdomains may reflect structural features that enhance pathogen uptake either by providing high-avidity platforms and/or by protecting against rapid microdomain endocytosis. PMID- 21641312 TI - Preparation and properties of asymmetric large unilamellar vesicles: interleaflet coupling in asymmetric vesicles is dependent on temperature but not curvature. AB - Asymmetry of inner and outer leaflet lipid composition is an important characteristic of eukaryotic plasma membranes. We previously described a technique in which methyl-beta-cyclodextrin-induced lipid exchange is used to prepare biological membrane-like asymmetric small unilamellar vesicles (SUVs). Here, to mimic plasma membranes more closely, we used a lipid-exchange-based method to prepare asymmetric large unilamellar vesicles (LUVs), which have less membrane curvature than SUVs. Asymmetric LUVs in which sphingomyelin (SM) or SM + 1-palmitoyl-2-oleoyl-phosphatidylcholine was exchanged into the outer leaflet of vesicles composed of 1,2-dioleoyl-phosphatidylethanolamine (DOPE) and 1-palmitoyl 2-oleoyl-phosphatidylserine (POPS) were prepared with or without cholesterol. Approximately 80-100% replacement of outer leaflet DOPE and POPS was achieved. At room temperature, SM exchange into the outer leaflet increased the inner leaflet lipid order, suggesting significant interleaflet interaction. However, the SM rich outer leaflet formed an ordered state, melting with a midpoint at ~37 degrees C. This was about the same value observed in pure SM vesicles, and was significantly higher than that observed in symmetric vesicles with the same SM content, which melted at ~20 degrees C. In other words, ordered state formation by outer-leaflet SM in asymmetric vesicles was not destabilized by an inner leaflet composed of DOPE and POPS. These properties suggest that the coupling between the physical states of the outer and inner leaflets in these asymmetric LUVs becomes very weak as the temperature approaches 37 degrees C. Overall, the properties of asymmetric LUVs were very similar to those previously observed in asymmetric SUVs, indicating that they do not arise from the high membrane curvature of asymmetric SUVs. PMID- 21641313 TI - Cooperative [Ca2+]-dependent regulation of the rate of myosin binding to actin: solution data and the tropomyosin chain model. AB - The regulation of muscle contraction by calcium involves interactions among actin filaments, myosin-S1, tropomyosin (Tm), and troponin (Tn). We have extended our previous model in which the TmTn regulatory units are treated as a continuous flexible chain, and applied it to transient kinetic data. We have measured the time course of myosin-S1 binding to actin-Tm-Tn filaments in solution at various calcium levels with [actin]/[myosin] ratios of 10 and 0.1, which exhibit modest slowing as [Ca(2+)] is reduced and a lag phase at low calcium. These observations can be explained if myosin binds to actin in two steps, where the first step is rate-limiting and blocked by TmTnI at low calcium, and the second step is fast, reversible, and controlled by the neighboring configuration of coupled tropomyosin-troponin units. The model can describe the calcium dependence of the observed myosin binding reactions and predicts cooperative calcium binding to TnC with competition between actin and Ca-TnC for the binding of TnI. Implications for theories of thin-filament regulation in muscle are discussed. PMID- 21641314 TI - An electrostatic model with weak actin-myosin attachment resolves problems with the lattice stability of skeletal muscle. AB - The stability of the filament lattice in relaxed striated muscle can be viewed as a balance of electrostatic and van der Waals forces. The simplest electrostatic model, where actin and myosin filaments are treated as charged cylinders, generates reasonable lattice spacings for skinned fibers. However, this model predicts excessive radial stiffness under osmotic pressure and cannot account for the initial pressure (~1 kPa) required for significant compression. Good agreement with frog compression data is obtained with an extended model, in which S1 heads are weakly attached to actin when the lattice spacing is reduced below a critical value; further compression moves fixed negative charges on the heads closer to the myofilament backbone as they attach at a more acute angle to actin. The model predicts pH data in which the lattice shrinks as pH is lowered and protons bind to filaments. Electrostatic screening implies that the lattice shrinks with increasing ionic strength, but the observed expansion of the frog lattice at ionic strengths above 0.1 M with KCl might be explained if Cl(-) binds to sites on the motor domain of S1. With myosin-myosin and actin-actin interactions, the predicted lattice spacing decreases slightly with sarcomere length, with a more rapid decrease when actin-myosin filament overlap is very small. PMID- 21641315 TI - Reconstitution of contractile actomyosin bundles. AB - Contractile actomyosin bundles are critical for numerous aspects of muscle and nonmuscle cell physiology. Due to the varying composition and structure of actomyosin bundles in vivo, the minimal requirements for their contraction remain unclear. Here, we demonstrate that actin filaments and filaments of smooth muscle myosin motors can self-assemble into bundles with contractile elements that efficiently transmit actomyosin forces to cellular length scales. The contractile and force-generating potential of these minimal actomyosin bundles is sharply sensitive to the myosin density. Above a critical myosin density, these bundles are contractile and generate large tensile forces. Below this threshold, insufficient cross-linking of F-actin by myosin thick filaments prevents efficient force transmission and can result in rapid bundle disintegration. For contractile bundles, the rate of contraction decreases as forces build and stalls under loads of ~0.5 nN. The dependence of contraction speed and stall force on bundle length is consistent with bundle contraction occurring by several contractile elements connected in series. Thus, contraction in reconstituted actomyosin bundles captures essential biophysical characteristics of myofibrils while lacking numerous molecular constituents and structural signatures of sarcomeres. These results provide insight into nonsarcomeric mechanisms of actomyosin contraction found in smooth muscle and nonmuscle cells. PMID- 21641316 TI - Striated acto-myosin fibers can reorganize and register in response to elastic interactions with the matrix. AB - The remarkable striation of muscle has fascinated many for centuries. In developing muscle cells, as well as in many adherent, nonmuscle cell types, striated, stress fiberlike structures with sarcomere-periodicity tend to register: Based on several studies, neighboring, parallel fibers at the basal membrane of cultured cells establish registry of their respective periodic sarcomeric architecture, but, to our knowledge, the mechanism has not yet been identified. Here, we propose for cells plated on an elastic substrate or adhered to a neighboring cell, that acto-myosin contractility in striated fibers close to the basal membrane induces substrate strain that gives rise to an elastic interaction between neighboring striated fibers, which in turn favors interfiber registry. Our physical theory predicts a dependence of interfiber registry on externally controllable elastic properties of the substrate. In developing muscle cells, registry of striated fibers (premyofibrils and nascent myofibrils) has been suggested as one major pathway of myofibrillogenesis, where it precedes the fusion of neighboring fibers. This suggests a mechanical basis for the optimal myofibrillogenesis on muscle-mimetic elastic substrates that was recently observed by several groups in cultures of mouse-, human-, and chick-derived muscle cells. PMID- 21641317 TI - Propulsive forces on the flagellum during locomotion of Chlamydomonas reinhardtii. AB - The distributed propulsive forces exerted on the flagellum of the swimming alga Chlamydomonas reinhardtii by surrounding fluid were estimated from experimental image data. Images of uniflagellate mutant Chlamydomonas cells were obtained at 350 frames/s with 125-nm spatial resolution, and the motion of the cell body and the flagellum were analyzed in the context of low-Reynolds-number fluid mechanics. Wild-type uniflagellate cells, as well as uniflagellate cells lacking inner dynein arms (ida3) or outer dynein arms (oda2) were studied. Ida3 cells exhibit stunted flagellar waveforms, whereas oda2 cells beat with lower frequency. Image registration and sorting algorithms provided high-resolution estimates of the motion of the cell body, as well as detailed kinematics of the flagellum. The swimming cell was modeled as an ellipsoid in Stokes flow, propelled by viscous forces on the flagellum. The normal and tangential components of force on the flagellum (f(N) and f(T)) were related by resistive coefficients (C(N) and C(T)) to the corresponding components of velocity (V(N) and V(T)).The values of these coefficients were estimated by satisfying equilibrium requirements for force and torque on the cell. The estimated values of the resistive coefficients are consistent among all three genotypes and similar to theoretical predictions. PMID- 21641318 TI - Linker histones incorporation maintains chromatin fiber plasticity. AB - Genomic DNA in eukaryotic cells is organized in supercoiled chromatin fibers, which undergo dynamic changes during such DNA metabolic processes as transcription or replication. Indeed, DNA-translocating enzymes like polymerases produce physical constraints in vivo. We used single-molecule micromanipulation by magnetic tweezers to study the response of chromatin to mechanical constraints in the same range as those encountered in vivo. We had previously shown that under positive torsional constraints, nucleosomes can undergo a reversible chiral transition toward a state of positive topology. We demonstrate here that chromatin fibers comprising linker histones present a torsional plasticity similar to that of naked nucleosome arrays. Chromatosomes can undergo a reversible chiral transition toward a state of positive torsion (reverse chromatosome) without loss of linker histones. PMID- 21641319 TI - Measuring the folding landscape of a harmonically constrained biopolymer. AB - Pioneering studies have shown that the probability distribution of opening length for a DNA hairpin, recorded under constant force using an optical trap, can be used to reconstruct the energy landscape of the transition. However, measurements made under constant force are subject to some limitations. Under constant force a system with a sufficiently high energy barrier spends most of its time in the closed or open conformation, with relatively few statistics collected in the transition state region. We describe a measurement scheme in which the system is driven progressively through the transition by an optical trap and an algorithm is used to extract the energy landscape of the transition from the fluctuations recorded during this process. We illustrate this technique in simulations and demonstrate its effectiveness in experiments on a DNA hairpin. We find that the combination of this technique with the use of short DNA handles facilitates a high-resolution measurement of the hairpin's folding landscape with a very short measurement time. PMID- 21641320 TI - Theory for RNA folding, stretching, and melting including loops and salt. AB - Secondary structure formation of nucleic acids strongly depends on salt concentration and temperature. We develop a theory for RNA folding that correctly accounts for sequence effects, the entropic contributions associated with loop formation, and salt effects. Using an iterative expression for the partition function that neglects pseudoknots, we calculate folding free energies and minimum free energy configurations based on the experimentally derived basepairing free energies. The configurational entropy of loop formation is modeled by the asymptotic expression -clnm, where m is the length of the loop and c the loop exponent, which is an adjustable constant. Salt effects enter in two ways: first, we derive salt-induced modifications of the free energy parameters for describing basepairing, and second, we include the electrostatic free energy for loop formation. Both effects are modeled on the Debye-Huckel level including counterion condensation. We validate our theory for two different RNA sequences. For tRNA-phe, the resultant heat capacity curves for thermal denaturation at various salt concentrations accurately reproduce experimental results. For the P5ab RNA hairpin, we derive the global phase diagram in the three-dimensional space spanned by temperature, stretching force, and salt concentration and obtain good agreement with the experimentally determined critical unfolding force. We show that for a proper description of RNA melting and stretching, both salt and loop entropy effects are needed. PMID- 21641321 TI - Molecular dynamics simulations of DNA/PEI complexes: effect of PEI branching and protonation state. AB - Complexes formed by DNA and polyethylenimine (PEI) are of great research interest because of their application in gene therapy. In this work, we carried out all atom molecular dynamics simulations to study eight types of DNA/PEI complexes, each of which was formed by one DNA duplex d(CGCGAATTCGCG)(2) and one PEI. We used eight different PEIs with four different degrees of branching and two protonation ratios of amine groups (23% and 46%) in the simulations to investigate how the branching degree and protonation state can affect the binding. We found that 46% protonated PEIs form more stable complexes with DNA, and the binding is achieved mainly through direct interaction between the protonated amine groups on PEI and the electronegative oxygens on the DNA backbone, with some degree of interaction with electronegative groove nitrogens/oxygens. For the 23% protonated PEIs, indirect interaction mediated by one or more water molecules plays an important role in binding. Compared with the protonation state, the degree of branching has a smaller effect on binding, which essentially diminishes at the protonation ratio of 46%. These simulations shed light on the detailed mechanism(s) of PEI binding to DNA, and may facilitate the design of PEI-based gene delivery carriers. PMID- 21641322 TI - Structural basis of folding cooperativity in model proteins: insights from a microcanonical perspective. AB - Two-state cooperativity is an important characteristic in protein folding. It is defined by a depletion of states that lie energetically between folded and unfolded conformations. There are different ways to test for two-state cooperativity; however, most of these approaches probe indirect proxies of this depletion. Generalized-ensemble computer simulations allow us to unambiguously identify this transition by a microcanonical analysis on the basis of the density of states. Here, we present a detailed characterization of several helical peptides obtained by coarse-grained simulations. The level of resolution of the coarse-grained model allowed to study realistic structures ranging from small alpha-helices to a de novo three-helix bundle without biasing the force field toward the native state of the protein. By linking thermodynamic and structural features, we are able to show that whereas short alpha-helices exhibit two-state cooperativity, the type of transition changes for longer chain lengths because the chain forms multiple helix nucleation sites, stabilizing a significant population of intermediate states. The helix bundle exhibits signs of two-state cooperativity owing to favorable helix-helix interactions, as predicted from theoretical models. A detailed analysis of secondary and tertiary structure formation fits well into the framework of several folding mechanisms and confirms features that up to now have been observed only in lattice models. PMID- 21641323 TI - Location trumps length: polyglutamine-mediated changes in folding and aggregation of a host protein. AB - Expanded CAG diseases are progressive neurodegenerative disorders in which specific proteins have an unusually long polyglutamine stretch. Although these proteins share no other sequence or structural homologies, they all aggregate into intracellular inclusions that are believed to be pathological. We sought to determine what impact the position and number of glutamines have on the structure and aggregation of the host protein, apomyoglobin. Variable-length polyQ tracts were inserted either into the loop between the C- and D-helices (Q(n)CD) or at the N-terminus (Q(n)NT). The Q(n)CD mutants lost some alpha-helix and gained unordered and/or beta-sheet in a length-dependent manner. These mutants were partially unfolded and rapidly assembled into soluble chain-like oligomers. In sharp contrast, the Q(n)NT mutants largely retained wild-type tertiary structure but associated into long, fibrillar aggregates. Control proteins with glycine serine repeats (GS(8)CD and GS(8)NT) were produced. GS(8)CD exhibited similar structural perturbations and aggregation characteristics to an analogously sized Q(16)CD, indicating that the observed effects are independent of amino acid composition. In contrast to Q(16)NT, GS(8)NT did not form fibrillar aggregates. Thus, soluble oligomers are produced through structural perturbation and do not require polyQ, whereas classic fibrils arise from specific polyQ intermolecular interactions in the absence of misfolding. PMID- 21641324 TI - Perturbation of the stability of amyloid fibrils through alteration of electrostatic interactions. AB - The self-assembly of proteins and peptides into polymeric amyloid fibrils is a process that has important implications ranging from the understanding of protein misfolding disorders to the discovery of novel nanobiomaterials. In this study, we probe the stability of fibrils prepared at pH 2.0 and composed of the protein insulin by manipulating electrostatic interactions within the fibril architecture. We demonstrate that strong electrostatic repulsion is sufficient to disrupt the hydrogen-bonded, cross-beta network that links insulin molecules and ultimately results in fibril dissociation. The extent of this dissociation correlates well with predictions for colloidal models considering the net global charge of the polypeptide chain, although the kinetics of the process is regulated by the charge state of a single amino acid. We found the fibrils to be maximally stable under their formation conditions. Partial disruption of the cross-beta network under conditions where the fibrils remain intact leads to a reduction in their stability. Together, these results support the contention that a major determinant of amyloid stability stems from the interactions in the structured core, and show how the control of electrostatic interactions can be used to characterize the factors that modulate fibril stability. PMID- 21641325 TI - Isolating toxic insulin amyloid reactive species that lack beta-sheets and have wide pH stability. AB - Amyloid diseases, including Alzheimer's disease, are characterized by aggregation of normally functioning proteins or peptides into ordered, beta-sheet rich fibrils. Most of the theories on amyloid toxicity focus on the nuclei or oligomers in the fibril formation process. The nuclei and oligomers are transient species, making their full characterization difficult. We have isolated toxic protein species that act like an oligomer and may provide the first evidence of a stable reactive species created by disaggregation of amyloid fibrils. This reactive species was isolated by dissolving amyloid fibrils at high pH and it has a mass >100 kDa and a diameter of 48 +/- 15 nm. It seeds the formation of fibrils in a dose dependent manner, but using circular dichroism and deep ultraviolet resonance Raman spectroscopy, the reactive species was found to not have a beta sheet rich structure. We hypothesize that the reactive species does not decompose at high pH and maintains its structure in solution. The remaining disaggregated insulin, excluding the toxic reactive species that elongated the fibrils, returned to native structured insulin. This is the first time, to our knowledge, that a stable reactive species of an amyloid reaction has been separated and characterized by disaggregation of amyloid fibrils. PMID- 21641326 TI - Photoreactions of aureochrome-1. AB - Aureochrome is a recently discovered blue light photosensor that controls a light dependent morphology change. As a photosensor, it has a unique DNA binding domain (bZIP). Although the biological functions of aureochrome have been revealed, the fundamental photochemistry of this protein has not been elucidated. The photochemical reaction dynamics of the LOV (light, oxygen, or voltage) domain of aureochrome-1 (AUREO1-LOV) and the LOV domain with the bZIP domain (AUREO1-ZL) were studied by employing the transient-grating (TG) technique, using size exclusion chromatography to verify results. For both samples, adduct formation takes place with a time constant of 2.8 MUs. Although significant diffusion changes were observed for both AUREO1-LOV and AUREO1-ZL after adduct formation, the origins of these changes were significantly different. The TG signal of AUREO1-LOV was strongly concentration-dependent. From analysis of the signal, it was concluded that AUREO1-LOV exists in equilibrium between the monomer and dimer, and dimerization of the monomer is the main reaction, i.e., irradiation with blue light enhances the strength of the interdomain interaction. On the other hand, the reaction of AUREO1-ZL is independent of concentration, suggesting that an intraprotein conformational change occurs in the bZIP domain with a time constant of 160 ms. These results revealed the different reactions and roles of the two domains; the LOV domain acts as a photosensor, leading to a subsequent conformational change in the bZIP domain, which should change its ability to bind to DNA. A model is proposed that demonstrates how aureochrome uses blue light to control its affinity for DNA. PMID- 21641327 TI - Single-shot optical sectioning using two-color probes in HiLo fluorescence microscopy. AB - We describe a wide-field fluorescence microscope setup which combines HiLo microscopy technique with the use of a two-color fluorescent probe. It allows one shot fluorescence optical sectioning of thick biological moving sample which is illuminated simultaneously with a flat and a structured pattern at two different wavelengths. Both homogenous and structured fluorescence images are spectrally separated at detection and combined similarly with the HiLo microscopy technique. We present optically sectioned full-field images of Xenopus laevis embryos acquired at 25 images/s frame rate. PMID- 21641328 TI - Tracking single particles and elongated filaments with nanometer precision. AB - Recent developments in image processing have greatly advanced our understanding of biomolecular processes in vitro and in vivo. In particular, using Gaussian models to fit the intensity profiles of nanometer-sized objects have enabled their two-dimensional localization with a precision in the one-nanometer range. Here, we present an algorithm to precisely localize curved filaments whose structures are characterized by subresolution diameters and micrometer lengths. Using surface-immobilized microtubules, fluorescently labeled with rhodamine, we demonstrate positional precisions of ~2 nm when determining the filament centerline and ~9 nm when localizing the filament tips. Combined with state-of the-art single particle tracking we apply the algorithm 1), to motor-proteins stepping on immobilized microtubules, 2), to depolymerizing microtubules, and 3), to microtubules gliding over motor-coated surfaces. PMID- 21641329 TI - Minor complexes at work: light-harvesting by carotenoids in the photosystem II antenna complexes CP24 and CP26. AB - Plant photosynthesis relies on the capacity of chlorophylls and carotenoids to absorb light. One of the roles of carotenoids is to harvest green-blue light and transfer the excitation energy to the chlorophylls. The corresponding dynamics were investigated here for the first time, to our knowledge, in the CP26 and CP24 minor antenna complexes. The results for the two complexes differ substantially. In CP26 fast transfer (80 fs) occurs from the carotenoid S(2) state to chlorophylls a absorbing at 675 and 678 nm, whereas transfer from the hot S(1) state to the lowest energy chlorophylls is observed in <1 ps. In CP24, energy transfer from the S(2) state leads in 80 fs to the population of chlorophylls b and high-energy chlorophylls a absorbing at 670 nm, whereas the low-energy chlorophylls a are populated only in several picoseconds. The results suggest that CP26 has a structural and functional organization similar to that of LHCII, whereas CP24 differs substantially from the other Lhc complexes, especially regarding the lutein L1 binding domain. No energy transfer from the carotenoid S(1) state to chlorophylls was observed in either complex, suggesting that this state is energetically below the chlorophyll Qy state and therefore may play a role in the quenching of chlorophyll excitations. PMID- 21641330 TI - Spot variation fluorescence correlation spectroscopy allows for superresolution chronoscopy of confinement times in membranes. AB - Resolving the dynamical interplay of proteins and lipids in the live-cell plasma membrane represents a central goal in current cell biology. Superresolution concepts have introduced a means of capturing spatial heterogeneity at a nanoscopic length scale. Similar concepts for detecting dynamical transitions (superresolution chronoscopy) are still lacking. Here, we show that recently introduced spot-variation fluorescence correlation spectroscopy allows for sensing transient confinement times of membrane constituents at dramatically improved resolution. Using standard diffraction-limited optics, spot-variation fluorescence correlation spectroscopy captures signatures of single retardation events far below the transit time of the tracer through the focal spot. We provide an analytical description of special cases of transient binding of a tracer to pointlike traps, or association of a tracer with nanodomains. The influence of trap mobility and the underlying binding kinetics are quantified. Experimental approaches are suggested that allow for gaining quantitative mechanistic insights into the interaction processes of membrane constituents. PMID- 21641331 TI - Probing rotational viscosity in synaptic vesicles. AB - The synaptic vesicle (SV) is a central organelle in neurotransmission, and previous studies have suggested that SV protein 2 (SV2) may be responsible for forming a gel-like matrix within the vesicle. Here we measured the steady-state rotational anisotropy of the fluorescent dye, Oregon Green, within individual SVs. By also measuring the fluorescence lifetime of Oregon Green in SVs, we determined the mean rotational viscosity to be 16.49 +/- 0.12 cP for wild-type (WT) empty mice vesicles (i.e., with no neurotransmitters), 11.21 +/- 0.12 cP for empty vesicles from SV2 knock-out mice, and 11.40 +/- 0.65 cP for WT mice vesicles loaded with the neurotransmitter glutamate (Glu). This measurement shows that SV2 is an important determinant of viscosity within the vesicle lumen, and that the viscosity decreases when the vesicles are filled with Glu. The viscosities of both empty SV2 knock-out vesicles and Glu-loaded WT vesicles were significantly different from that of empty WT SVs (p < 0.05). This measurement represents the smallest enclosed volume in which rotational viscosity has been measured thus far. PMID- 21641332 TI - Molecular mechanisms of production and scavenging of reactive oxygen species by photosystem II. AB - Photosystem II (PSII) is a multisubunit protein complex in cyanobacteria, algae and plants that use light energy for oxidation of water and reduction of plastoquinone. The conversion of excitation energy absorbed by chlorophylls into the energy of separated charges and subsequent water-plastoquinone oxidoreductase activity are inadvertently coupled with the formation of reactive oxygen species (ROS). Singlet oxygen is generated by the excitation energy transfer from triplet chlorophyll formed by the intersystem crossing from singlet chlorophyll and the charge recombination of separated charges in the PSII antenna complex and reaction center of PSII, respectively. Apart to the energy transfer, the electron transport associated with the reduction of plastoquinone and the oxidation of water is linked to the formation of superoxide anion radical, hydrogen peroxide and hydroxyl radical. To protect PSII pigments, proteins and lipids against the oxidative damage, PSII evolved a highly efficient antioxidant defense system comprising either a non-enzymatic (prenyllipids such as carotenoids and prenylquinols) or an enzymatic (superoxide dismutase and catalase) scavengers. It is pointed out here that both the formation and the scavenging of ROS are controlled by the energy level and the redox potential of the excitation energy transfer and the electron transport carries, respectively. The review is focused on the mechanistic aspects of ROS production and scavenging by PSII. This article is part of a Special Issue entitled: Photosystem II. PMID- 21641333 TI - FTIR spectroscopy of the reaction center of Chloroflexus aurantiacus: photoreduction of the bacteriopheophytin electron acceptor. AB - Mid-infrared spectral changes associated with the photoreduction of the bacteriopheophytin electron acceptor H(A) in reaction centers (RCs) of the filamentous anoxygenic phototrophic bacterium Chloroflexus (Cfl.) aurantiacus are examined by light-induced Fourier transform infrared (FTIR) spectroscopy. The light-induced H(A)(-)/H(A) FTIR (1800-1200cm(-1)) difference spectrum of Cfl. aurantiacus RCs is compared to that of the previously well characterized purple bacterium Rhodobacter (Rba.) sphaeroides RCs. The most notable feature is that the large negative IR band at 1674cm(-1) in Rba. sphaeroides R-26, attributable to the loss of the absorption of the 13(1)-keto carbonyl of H(A) upon the radical anion H(A)(-) formation, exhibits only a very minor upshift to 1675cm(-1) in Cfl. aurantiacus. In contrast, the absorption band of the 131-keto C=O of H(A)(-) is strongly upshifted in the spectrum of Cfl. aurantiacus compared to that of Rba. sphaeroides (from 1588 to 1623cm(-1)). The data are discussed in terms of: (i) replacing the glutamic acid at L104 in Rba. sphaeroides R-26 RCs by a weaker hydrogen bond donor, a glutamine, at the equivalent position L143 in Cfl. aurantiacus RCs; (ii) a strengthening of the hydrogen-bonding interaction of the 131-keto C=O of H(A) with Glu L104 and Gln L143 upon H(A)(-) formation and (iii) a possible influence of the protein dielectric environment on the 131-keto C=O stretching frequency of neutral H(A). A conformational heterogeneity of the 133 ester C=O group of H(A) is detected for Cfl. aurantiacus RCs similar to what has been previously described for purple bacterial RCs. PMID- 21641334 TI - Investigating the cationic side chains of the antimicrobial peptide tritrpticin: hydrogen bonding properties govern its membrane-disruptive activities. AB - The positively charged side chains of cationic antimicrobial peptides are generally thought to provide the initial long-range electrostatic attractive forces that guide them towards the negatively charged bacterial membranes. Peptide analogs were designed to examine the role of the four Arg side chains in the cathelicidin peptide tritrpticin (VRRFPWWWPFLRR). The analogs include several noncoded Arg and Lys derivatives that offer small variations in side chain length and methylation state. The peptides were tested for bactericidal and hemolytic activities, and their membrane insertion and permeabilization properties were characterized by leakage assays and fluorescence spectroscopy. A net charge of +5 for most of the analogs maintains their high antimicrobial activity and directs them towards preferential insertion into model bacterial membrane systems with a similar extent of burial of the Trp side chains. However the peptides exhibit significant functional differences. Analogs with methylated cationic side chains cause lower levels of membrane leakage and are associated with lower hemolytic activities, making them potentially attractive pharmaceutical candidates. Analogs containing the Arg guanidinium groups cause more membrane disruption than those containing the Lys amino groups. Peptides in the latter group with shorter side chains have increased membrane activity and conversely, elongating the Arg residue causes slightly higher membrane activity. Altogether, the potential for strong hydrogen bonding between the four positive Arg side chains with the phospholipid head groups seems to be a determinant for the membrane disruptive properties of tritrpticin and many related cationic antimicrobial peptides. PMID- 21641335 TI - Glioma-derived mutations in isocitrate dehydrogenase 2 beneficial to traditional chemotherapy. AB - Heterozygous mutations in either the R132 residue of isocitrate dehydrogenase I (IDH1) or the R172 residue of IDH2 in human gliomas were recently highlighted. In the present study, we report that mutations of IDH1 and IDH2 are not detected in the rat C6 glioma cell line model, which suggests that these mutations are not required for the development of glioblastoma induced by N,N'-nitroso-methylurea. The effects of IDH2 and IDH2(R172G) on C6 cells proliferation and sensitivity to chemotherapy and the possible mechanism are analyzed at the cellular level. IDH1 and IDH2 mutations lead to simultaneous loss and gain of activities in the production of alpha-ketoglutarate (alpha-KG) and 2-hydroxyglutarate (2HG), respectively, and result in lowering NADPH levels even further. The low NADPH levels can sensitize tumors to chemotherapy, and account for the prolonged survival of patients harboring the mutations. Our data extrapolate potential importance of the in vitro rat C6 glioma cell model, show that the IDH2(R172G) mutation in gliomas may give a benefit to traditional chemotherapy of this cancer and serve as an important complement to existing research on this topic. PMID- 21641336 TI - Transcription factor COUP-TFII is indispensable for venous and lymphatic development in zebrafish and Xenopus laevis. AB - Transcription factors play a central role in cell fate determination. Gene targeting in mice revealed that Chicken Ovalbumin Upstream Promoter-Transcription Factor II (COUP-TFII, also known as Nuclear Receptor 2F2 or NR2F2) induces a venous phenotype in endothelial cells (ECs). More recently, NR2F2 was shown to be required for initiating the expression of Prox1, responsible for lymphatic commitment of venous ECs. Small animal models like zebrafish embryos and Xenopus laevis tadpoles have been very useful to elucidate mechanisms of (lymph) vascular development. Therefore, the role of NR2F2 in (lymph) vascular development was studied by eliminating its expression in these models. Like in mice, absence of NR2F2 in zebrafish resulted in distinct vascular defects including loss of venous marker expression, major trunk vessel fusion and vascular leakage. Both in zebrafish and Xenopus the development of the main lymphatic structures was severely hampered. NR2F2 knockdown significantly decreased prox1 expression in zebrafish ECs and the same manipulation affected lymphatic (L)EC commitment, migration and function in Xenopus tadpoles. Therefore, the role of NR2F2 in EC fate determination is evolutionary conserved. PMID- 21641337 TI - Importance of cholesterol and oxysterols metabolism in the pharmacology of tamoxifen and other AEBS ligands. AB - Tamoxifen is one of the major drugs used for the hormonotherapy of estrogen receptor positive breast cancers. However, its therapeutic efficacy can be limited by acquired resistance and tumor recurrence can occur after several years of treatment. Tamoxifen is known as the prototypical modulator of estrogen receptors, but other targets have been identified that could account for its pharmacology. In particular, tamoxifen binds with high affinity to the microsomal antiestrogen binding site (AEBS) and inhibits cholesterol esterification at therapeutic doses. We have recently shown that the AEBS was a hetero-oligomeric complex composed of 3beta-hydroxysterol-Delta(8)-Delta(7)-isomerase and 3beta hydroxysterol-Delta(7)-reductase, that binds different structural classes of ligands, including selective estrogen receptor modulators, several sigma receptor ligands, poly-unsaturated fatty acids and ring B oxysterols. We established a link between the modulation of cholesterol metabolism by tamoxifen and other AEBS ligands and their capacity to induce breast cancer cell differentiation, apoptosis and autophagy. Moreover, we showed that the AEBS carries out cholesterol-5,6-epoxide hydrolase activity and established that cholesterol-5,6 epoxide hydrolase is a new target for tamoxifen and other AEBS ligands. Finally in this review, we report on recent data from the literature showing how the modulation of cholesterol and oxysterol metabolism can be linked to the antitumor and chemopreventive properties of tamoxifen, and give new perspectives to improve the clinical outcome of the hormonotherapy of breast cancers. PMID- 21641338 TI - Was a recent manufacturer recall of CA-125 reagents necessary? PMID- 21641339 TI - Method-dependent HbA1c values in a family with hemoglobin Himeji. AB - BACKGROUND: Hb Himeji is variant hemoglobin associated with increased glycation in a mutated beta chain. We measured HbA1c using various methods in a family with Hb Himeji. METHODS: The proband was a 42-y female. While receiving treatment for Graves' disease, an oral glucose tolerance test showed normal glucose tolerance, but HbA1c by enzymatic assay was abnormally elevated (11.6%). Hemoglobin gene analysis identified Hb Himeji [beta140 (H18) Ala->Asp]. RESULTS: HbA1c values measured by high-performance liquid chromatography (HPLC; HLC-723G8 and HA-8160 instruments), immunoassay, enzymatic assay, affinity method, and electrospray ionization/mass spectrometry were 3.2%, 5.2%, 11.5%, 9.7%, 7.2%, and 9.6%, respectively. Glycation product of the variant hemoglobin measured by HPLC, using HLC-723G8 and HA-8160, was 9.1% and 4.5%, respectively. The proband's father with type 2 diabetes was the first reported case of Hb Himeji. HbA1c by affinity method was markedly elevated (18.0%), but it was 5.3% by HPLC. The proband's two sisters also had Hb Himeji variant and similar method-dependent discrepancies in HbA1c values were observed. CONCLUSIONS: In the patients with Hb Himeji, discrepancies occur between plasma glucose and HbA1c with all measurement methods because of differences in HPLC mobility, increased glycation, and antigenic changes of the variant beta chain. PMID- 21641340 TI - Rex1 (Zfp42) null mice show impaired testicular function, abnormal testis morphology, and aberrant gene expression. AB - Rex1 (Zfp42), GeneID 132625, is a gene whose expression is closely associated with pluripotency/multipotency in both mouse and human embryonic stem cells. To study the function of the murine Rex1 gene in vivo, we have used cre/lox technology to create Rex1(floxed) mice and mice deficient in Rex1 gene function. Rex1(-/-)males are characterized by an age-associated decrease in sperm counts, abnormal sperm morphology, and mild testicular atrophy. We characterized global patterns of gene expression in primary germ cells by microarray and identified the growth hormone responsive gene, GRTP1, as a transcript present at a 4.5 fold higher level in wild type (WT) compared to Rex1(-/-) mice. We analyzed immature germ cell (Dazl), proliferating (PCNA), and Sertoli cell populations, and quantitated levels of apoptosis in Rex1(-/-) as compared to WT testes. We evaluated the expression of proteins previously reported to correlate with Rex1 expression, such as STAT3, phospho-STAT3, p38, and phospho-p38 in the testis. We report a distinct cellular localization of total STAT3 protein in Rex1(-/-) affected testes. Our data suggest that loss of Rex1 leads to impaired testicular function. PMID- 21641341 TI - Rosuvastatin attenuates monocrotaline-induced pulmonary hypertension via regulation of Akt/eNOS signaling and asymmetric dimethylarginine metabolism. AB - This study was designed to investigate whether rosuvastatin could attenuate monocrotaline-induced pulmonary hypertension via regulation of Akt/eNOS signaling pathway and asymmetric dimethylarginine (ADMA) metabolism in rats. After a single dose injection of monocrotaline (60 mg/kg), oral administration of rosuvastatin (5mg/kg) was started from day 1 to day 28 (preventive administration) or from day 15 to day 28 (therapeutic administration), or with vehicle as corresponding controls. 28 days after monocrotaline, significant pulmonary hypertension characterized by pulmonary arterial medial wall thickening, right ventricular hypertrophy and right heart failure was observed. Rosuvastatin (5mg/kg, for 14 days and 28 days) treatment significantly attenuated monocrotaline-induced pulmonary vascular remodeling, right ventricular hypertrophy and dysfunction, and normalized the down-regulated pulmonary Akt/p-Akt and eNOS/p-eNOS expressions, while increased DDAH2 expression accompanied by decreased serum level of ADMA. However expression of PRMT1 and GSK3beta/p-GSK3beta did not differ among all groups (all P>0.05). We concluded that rosuvastatin inhibits monocrotaline induced pulmonary hypertension through normalization of Akt, eNOS and DDAH2 expressions, and decreasing the level of ADMA. PMID- 21641342 TI - Efficacy of aminaftone in a rat model of monocrotaline-induced pulmonary hypertension. AB - Pulmonary hypertension is characterized by increased vascular resistances, that could lead to right heart failure and death. Endothelin-1 (ET-1) is a peptide with strong vasoconstrictive and pro-fibrotic properties and is one of the main mediators of pulmonary hypertension. Aminaftone, a synthetic molecule derivative of 4-amynobenzoic acid, down-regulates ET-1 production in vitro by interfering with the transcription of the pre-pro-ET-1 gene. The aim of this study was to test whether the inhibition of ET-1 production by aminaftone attenuates the effects of pulmonary hypertension. Pulmonary hypertension was induced through s.c. injection of 60 mg/kg monocrotaline. The rats were randomly assigned to the following experimental groups: Control; Monocrotaline; Aminaftone 30 mg/kg/day; Aminaftone 150 mg/kg/day. After 5 weeks, mortality was significantly lower in the animals treated with aminaftone at both doses compared to monocrotaline alone. Aminaftone reduced plasma concentration of ET-1 and seemed to reduce right heart hypertrophy and the wall thickness of the pulmonary arteries at the highest dose. Aminaftone may represent a novel treatment strategy of pulmonary hypertension. PMID- 21641343 TI - Sulodexide prevents peripheral nerve damage in streptozotocin induced diabetic rats. AB - We investigated whether sulodexide has additional protective effects against peripheral nerve damage caused by microvascular dysfunction in a rat model of diabetes. Female Sprague-Dawley (SD) rats were divided into the following 4 groups (n=7-9/group): Normal, Normal+Sulodexide (sulodexide 10mg/kg), diabetic group, and diabetic+Sulodexide (sulodexide 10mg/kg). We assessed current perception threshold, skin blood flow, superoxide dismutase, and proteinuria in experimental rats after oral administration of sulodexide for 20 weeks. We also performed morphometric analysis of sciatic nerves and intraepidermal nerve fibers of the foot. Superoxide dismutase activity in the blood and sciatic nerve were increased significantly after sulodexide treatment in the diabetic group. Current perception threshold was reduced at 2000 Hz (633.3 +/- 24.15 vs 741.2 +/- 23.5 MUA, P<0.05) and skin blood flow was improved (10.90 +/- 0.67 vs 8.85 +/- 0.49 TPU, P<0.05) in the diabetic+Sulodexide group compared with the diabetic group. The mean myelinated axon area was significantly larger (56.6 +/- 2.2 vs 49.8 +/- 2.7 MUm(2), P<0.05) and the intraepidermal nerve fiber density was significantly less reduced (6.27 +/- 0.24 vs 5.40 +/- 0.25/mm, P<0.05) in the diabetic+Sulodexide group compared to the diabetic group. Our results demonstrate that sulodexide exhibits protective effects against peripheral nerve damage in a rat experimental model of diabetes. Therefore, these findings suggest that sulodexide is a potential new therapeutic agent for diabetic peripheral neuropathy. PMID- 21641344 TI - Projecting heart failure into bankruptcy in 2012? PMID- 21641346 TI - Potential impact of optimal implementation of evidence-based heart failure therapies on mortality. AB - BACKGROUND: Although multiple therapies have been shown to lower mortality in patients with heart failure (HF) and reduced left ventricular ejection fraction, their application in clinical practice has been less than ideal. To date, empiric estimation of the potential benefits that could be gained from eliminating these existing treatment gaps with optimal implementation has not been quantified. METHODS: Eligibility criteria for each evidence-based HF therapy, the estimated frequency of use/nonuse of specific treatments, the case fatality rates, and the risk reductions due to treatment were obtained from published sources. The numbers of deaths prevented or postponed because of each guideline-recommended therapy and overall were determined. RESULTS: Among patients with HF with reduced left ventricular ejection fraction in the United States (n = 2,644,800), the number eligible but not currently treated ranged from 139,749 for hydralazine/isorbide dinitrate to 852,512 for implantable cardioverter defibrillators. The comparative number of deaths that could potentially be prevented per year with optimal implementation of angiotensin-converting enzyme inhibitor/angiotensin receptor antagonist is 6,516; beta-blockers, 12,922; aldosterone antagonists, 21,407; hydralazine/isorbide dinitrate, 6,655; cardiac resynchronization therapy, 8,317; and implantable cardioverter defibrillators, 12,179. If these treatment benefits were additive, optimal implementation of all 6 therapies could potentially prevent 67,996 deaths a year. CONCLUSIONS: A substantial number of HF deaths in this country could potentially be prevented by optimal implementation of evidence-based therapies. These data may underscore the importance of performance improvement efforts to translate evidence-based therapy to routine clinical practice so as to reduce contemporary HF mortality. PMID- 21641347 TI - Prognostic value of cardiac troponin T in patients with moderate to severe heart failure scheduled for cardiac resynchronization therapy. AB - BACKGROUND: Predicting response to cardiac resynchronization therapy (CRT) is challenging. Highly sensitive cardiac troponin T (hsTnT) might predict response to CRT and identify patients at a high risk of experiencing severe cardiovascular events. We investigated whether baseline levels of hsTnT were associated with response to CRT and with severe cardiovascular events after long-term follow-up. METHODS: Eighty-one consecutive patients were included according to the current guidelines for CRT. Biochemical, functional, and clinical parameters were assessed at baseline and at 3, 6, and 12 months of follow-up; and mortality/cardiac transplantation after 46 +/- 6 months of follow-up was investigated. Cardiac magnetic resonance imaging and echocardiography were used to assess left ventricular function including viability and remodeling. RESULTS: Seventy-five patients completed 12 months of follow-up; and after a follow-up of 46 +/- 6 months, a total of 15 patients died, 13 of these from cardiovascular causes, and 7 underwent heart transplantation. Baseline hsTnT <15 ng/L predicted response to CRT and was associated with a more favorable outcome with regard to severe cardiovascular events. Multivariate analysis found that presence of transmural scar tissue/fibrosis on magnetic resonance imaging and use of statins were independently associated with higher concentrations of hsTnT at baseline. There was a strong correlation between hsTnT and N-terminal pro-B-type natriuretic peptide levels. CONCLUSIONS: Highly sensitive TnT levels were elevated in the majority of heart failure patients who were scheduled for CRT. The HsTnT levels predicted response to CRT as well as long-time survival. PMID- 21641345 TI - Rationale, design, and results from RENO-DEFEND 1: a randomized, dose-finding study of the selective A1 adenosine antagonist SLV320 in patients hospitalized with acute heart failure. AB - BACKGROUND: Baseline renal impairment as well as worsening renal function during hospitalization is associated with worse short- and long-term outcomes in patients hospitalized for acute heart failure (AHF). We hypothesized that selective A1 adenosine receptor blockade would induce natriuresis while preserving renal function in AHF patients with renal dysfunction. METHODS: A phase II, randomized, double-blind, placebo-controlled, parallel group, multicenter study to evaluate the efficacy and safety of 2.5, 7.5, 15, and 30 mg/d SLV320 (1 hour intravenous infusions of 1.25, 3.75, 7.5, and 15 mg SLV320, every 12 hours for 3 days [a total of 6 doses] in addition to standard therapy) in subjects hospitalized with AHF and renal impairment who meet all inclusion/exclusion criteria. The study planned to enroll 450 subjects, with 90 subjects allocated equally to each treatment arm. RESULTS: The study was terminated early. The decision, which was unrelated to the study conduct or results, with a total of 46 subjects randomized. Of those randomized, 6:8:8:8 and 6 patients, respectively, completed the study in each of the dosing subgroups, with placebo as the fifth group. For the 1.25-mg study group, the mean age was 73 years; mean (SD) systolic blood pressure (SBP), 128.5 (16.2); heart rate, 80.8 (25.0); brain natriuretic peptide, 969.7 (571.28); creatinine (MUmol/L), 149.7 (41.0); cystatin C, 1.468 (0.2777); estimated glomerular filtration rate, 33.8 (7.913); and blood urea nitrogen, 12.1 (2.9), with roughly similar values in each treatment arm. No seizures were reported during the study. Eight patients died during the study, none of whom were associated with the study drug per an independent, blinded, data safety monitoring board. CONCLUSION: Because of the limited number of subjects and variability observed in the results, no definite conclusions can be made regarding the efficacy and safety of SLV320. PMID- 21641349 TI - Time course of restoration of systolic and diastolic right ventricular function after pulmonary endarterectomy for chronic thromboembolic pulmonary hypertension. AB - BACKGROUND: In chronic thromboembolic pulmonary hypertension, right ventricular (RV) pressure overload causes RV remodeling and dysfunction. Successful pulmonary endarterectomy (PEA) initiates restoration of RV remodeling and global function. Little is known on the restoration of systolic and diastolic RV function. Using transthoracic echocardiography, we studied the time course and extent of postoperative restoration of systolic and diastolic RV function. METHODS: In chronic thromboembolic pulmonary hypertension (n = 55, 36 women, age 52 +/- 14 years), transthoracic echocardiography was performed before PEA (pre-PEA) and 2 weeks, 3 months, and 1 year postoperatively. RESULTS: Two weeks postoperatively, RV afterload and dimension had decreased significantly, without further improvement during follow-up. Global RV function, expressed by the myocardial performance index, showed a gradual improvement (from pre-PEA 0.58 +/- 0.29 to 0.45 +/- 0.38, 0.39 +/- 0.19, and 0.37 +/- 0.18). In contrast, 2 weeks after PEA systolic RV function, as assessed by tricuspid annular plane systolic velocity excursion and peak tricuspid annular systolic velocity of the RV, had worsened, with a subsequent incomplete restoration during follow-up: tricuspid annular plane systolic velocity excursion from 19.3 +/- 5.0 to 12.4 +/- 2.5, 15.3 +/- 3.0, and 16.8 +/- 2.9 mm and systolic velocity of the right ventricle from 11.4 +/- 3.0 to 9.6 +/- 2.0, 10.0 +/- 1.8, and 10.3 +/- 1.7 cm/s. Postoperative diastolic RV function also showed a biphasic response: tricuspid inflow-to annulus ratio from 6.1 +/- 3.0 to 9.5 +/- 3.5, 6.8 +/- 2.4, and 6.3 +/- 2.2 cm/s. Dynamics and ultimate level of restoration of systolic and diastolic RV function were similar in patients with and without residual pulmonary hypertension. CONCLUSIONS: Postoperative reduction in RV afterload caused an immediate improvement in RV dimension and global function. In contrast, systolic and diastolic RV function deteriorated after PEA with subsequently a gradual yet incomplete restoration during 1-year follow-up. PMID- 21641348 TI - Relationship of technetium-99m tetrofosmin-gated rest single-photon emission computed tomography myocardial perfusion imaging to death and hospitalization in heart failure patients: results from the nuclear ancillary study of the HF-ACTION trial. AB - BACKGROUND: We hypothesized that the severity of resting perfusion abnormalities assessed by the summed rest score (SRS) would be associated with a higher rate of adverse outcomes in patients with heart failure (HF) and reduced left ventricular (LV) ejection fraction (EF). METHODS: A subset of 240 subjects from HF-ACTION underwent resting technetium-99m tetrofosmin-gated single-photon emission computed tomography (SPECT) myocardial perfusion imaging (MPI). Images were evaluated using a 17-segment model to derive the SRS and additional nuclear variables. RESULTS: After adjusting for prespecified covariates, SRS was significantly associated with the primary end point (hazard ratio 0.98, 95% confidence interval [CI] 0.97-1.00, P = .04), with a higher SRS corresponding to lower risk of an event. This association was not present in the unadjusted analysis. The relationship between SRS and the primary outcome was likely due to a higher event ratein patients with ischemic HF and a low SRS. The LV phase SD was not predictive of the primary outcome (hazard ratio 1.00, 95% confidence interval 0.99-1.01, P = .49). In a post hoc analysis, nuclear variables provided incremental prognostic information when added to clinical information (P = .006). CONCLUSIONS: Gated SPECT MPI provides important information in patients with HF and reduced LVEF. In the adjusted analysis, SRS has an unexpected relationship with the primary end point. Phase SD was not associated with the primary end point. Rest-gated SPECT MPI provides incrementally greater prognostic information than clinical information alone. PMID- 21641350 TI - Depressive symptoms and inflammation in patients hospitalized for heart failure. AB - BACKGROUND: Depressive symptoms in patients with heart failure (HF) are common and might be associated with inflammation. No studies have examined both the cross-sectional and prospective association between inflammation and depressive symptoms in patients with HF with adequate correction for disease severity. The aim of this study was to describe if the cytokines interleukin-6 (IL-6) and C reactive protein (CRP) are associated with depressive symptoms in hospitalized HF patients. METHODS: Data from 517 patients hospitalized for HF from the COACH study were analyzed on inflammation markers (IL-6 and CRP) and depressive symptoms (Center for Epidemiological Studies Depression-Scale). RESULTS: Heart failure patients with depressive symptoms (n = 208, 40%) had significantly higher plasma values of IL-6 (median 12.8 pg/mL vs median 11.0 pg/mL, P = .018) and CRP (median 2.4 mg/mL vs median 2.1 mg/mL, P = .03) compared with the nondepressed patients. Structural equation modelling showed that the factor inflammation (including IL-6 and CRP) was associated with depressive symptoms (beta = 0.18, P < .05) when left ventricular ejection fraction and plasma values brain natriuretic peptides were included in the model. A small negative (beta = -0.18, P < .05) effect was found between inflammation at baseline and the change in depressive symptoms during the 18 months of follow-up. CONCLUSIONS: Higher levels of inflammatory markers are independently associated with depressive symptoms in HF patients, even after correcting for disease severity. There is no clear relationship between inflammation at baseline and depressive symptoms during the 18 months of follow-up. PMID- 21641351 TI - Site of latest activation in patients eligible for cardiac resynchronization therapy: patterns of dyssynchrony among different QRS configurations and impact of heart failure etiology. AB - INTRODUCTION: Cardiac resynchronization therapy (CRT) has emerged as a treatment option for patients with end-stage heart failure and a QRS duration >=120 ms. Nonetheless, many patients with a prolonged QRS do not demonstrate left ventricular (LV) mechanical dyssynchrony, and discrepancies between electrical and mechanical dyssynchrony have been observed. In addition, several studies demonstrated that superior benefits after CRT could be achieved when the LV pacing lead was positioned at the most delayed myocardial segment. METHODS: A total of 248 heart failure patients scheduled for CRT were included. In all patients, a 12-lead electrocardiogram and 2-dimensional echocardiogram were obtained. Patients were divided into 5 QRS configuration subgroups: narrow, left bundle-branch block, right bundle-branch block, intraventricular conduction delay, and right ventricular pacing. With speckle-tracking radial strain analysis, we evaluated time to peak radial strain. Next, the segments with the least and with the most mechanical activation delay were identified, and LV dyssynchrony was defined as the time delay between the two. RESULTS: Mean QRS duration was 164 +/- 31 ms. Mean LV dyssynchrony in all patients was 186 +/- 122 ms. Site of latest activation was predominantly located in the lateral (27%), posterior (26%), and inferior (20%) segments. Furthermore, extent of LV dyssynchrony was comparable between QRS configuration subgroups. An unequal distribution of LV segments with the most mechanical delay was observed in the left bundle-branch block and right ventricular pacing subgroups (P < .001 for both), whereas in the narrow, right bundle-branch block, and intraventricular conduction delay subgroups, a more homogeneous distribution was noted. No differences in distribution pattern or in extent of LV dyssynchrony were observed between ischemic and nonischemic heart failure patients. CONCLUSION: The lateral, posterior, and inferior segments take up 73% of the total latest activated segments in heart failure patients eligible for CRT. Presence of LV dyssynchrony can be observed in all QRS configurations. The site of latest activation may be outside the lateral or posterior segment, making echocardiographic assessment of LV dyssynchrony and site of latest activation a valuable technique to optimize patient outcome after CRT. PMID- 21641352 TI - Effects of tolvaptan on physician-assessed symptoms and signs in patients hospitalized with acute heart failure syndromes: analysis from the efficacy of vasopressin antagonism in heart failure outcome study with tolvaptan (EVEREST) trials. AB - BACKGROUND: A rapid and sustained relief of heart failure (HF) symptoms and signs is an important goal of management in patients hospitalized for acute HF syndromes (AHFS). To date, no novel therapy in AHFS have been shown to improve signs and symptoms throughout hospitalization. This study explores the clinical effects of tolvaptan, a vasopressin-2-receptor antagonist, in addition to standard medical therapies on physician-assessed signs and symptoms in hospitalized AHFS patients. METHODS: The EVEREST trial randomized 4,133 patients admitted with worsening HF and reduced ejection fraction (<= 40%) within 48 hours after hospital admission. On each inpatient day, investigators assessed dyspnea, orthopnea, fatigue, jugular venous distension (JVD), rales, and pedal edema by predefined ordinal scales. Responder analyses were performed for each sign and symptom, with significant clinical response defined as a change in one point on the measurement scale. RESULTS: Post hoc analysis demonstrated greater likelihood of clinical improvement in physician-assessed dyspnea, edema, orthopnea, and JVD among tolvaptan-treated subjects (P < .05) as early as inpatient day 1. This difference was observed throughout hospitalization only for JVD and orthopnea through day 3. CONCLUSION: The addition of tolvaptan to standard therapy for AHFS improves physician-assessed signs and symptoms during hospitalization without serious adverse short- or long-term effects. PMID- 21641353 TI - Late gadolinium enhancement on cardiac magnetic resonance and phenotypic expression in hypertrophic cardiomyopathy. AB - BACKGROUND: Cardiac magnetic resonance (CMR) imaging with late gadolinium enhancement (LGE) can identify areas of myocardial fibrosis in vivo in patients with hypertrophic cardiomyopathy (HCM). The aim of this study was to examine the association between clinical-morphological variables, risk factor for sudden death, and LGE findings in a consecutive, unselected population of HCM patients. METHODS: From January 2005 to August 2009, 124 HCM patients (53 +/- 17 years, 86 men) were prospectively evaluated with CMR examination, assessing left ventricular (LV) hypertrophy, function, and LGE. RESULTS: In univariate analysis, patients were divided into tertiles according to the number of segments positive for LGE (first tertile, 0.3 +/- 0.4; second tertile, 2.2 +/- 0.4; third tertile, 5.2 +/- 1.9 segments). Male gender (P = .05), maximum LV wall thickness (P = .002), nonsustained ventricular tachycardia (P = .001), ejection fraction <50% (P = .02), LV mass (P = .02), left atrium dilation (P = .04), perfusion defects (P <= .001), and telesystolic volume (P = .04) were all positively related with the number of segments of LGE. In multivariable analysis, male gender (P = .007), maximum LV wall thickness (P = .006), LV mass (P = .031), and perfusion alterations (P = .017) were independent predictors of LGE extent. CONCLUSIONS: Our study shows an independent association, even at multivariate analysis, between the entity of LGE and maximum LV wall thickness, mass, and perfusion defects in patients with HCM. Whether the presence and the extent of LGE translates into clinical events later on awaits further long-term follow-up studies. PMID- 21641354 TI - A randomized study of transendocardial injection of autologous bone marrow mononuclear cells and cell function analysis in ischemic heart failure (FOCUS HF). AB - BACKGROUND: Autologous bone marrow mononuclear cell (ABMMNC) therapy has shown promise in patients with heart failure (HF). Cell function analysis may be important in interpreting trial results. METHODS: In this prospective study, we evaluated the safety and efficacy of the transendocardial delivery of ABMMNCs in no-option patients with chronic HF. Efficacy was assessed by maximal myocardial oxygen consumption, single photon emission computed tomography, 2-dimensional echocardiography, and quality-of-life assessment (Minnesota Living with Heart Failure and Short Form 36). We also characterized patients' bone marrow cells by flow cytometry, colony-forming unit, and proliferative assays. RESULTS: Cell treated (n = 20) and control patients (n = 10) were similar at baseline. The procedure was safe; adverse events were similar in both groups. Canadian Cardiovascular Society angina score improved significantly (P = .001) in cell treated patients, but function was not affected. Quality-of-life scores improved significantly at 6 months (P = .009 Minnesota Living with Heart Failure and P = .002 physical component of Short Form 36) over baseline in cell-treated but not control patients. Single photon emission computed tomography data suggested a trend toward improved perfusion in cell-treated patients. The proportion of fixed defects significantly increased in control (P = .02) but not in treated patients (P = .16). Function of patients' bone marrow mononuclear cells was severely impaired. Stratifying cell results by age showed that younger patients (<=60 years) had significantly more mesenchymal progenitor cells (colony-forming unit fibroblasts) than patients >60 years (20.16 +/- 14.6 vs 10.92 +/- 7.8, P = .04). Furthermore, cell-treated younger patients had significantly improved maximal myocardial oxygen consumption (15 +/- 5.8, 18.6 +/- 2.7, and 17 +/- 3.7 mL/kg per minute at baseline, 3 months, and 6 months, respectively) compared with similarly aged control patients (14.3 +/- 2.5, 13.7 +/- 3.7, and 14.6 +/- 4.7 mL/kg per minute, P = .04). CONCLUSIONS: ABMMNC therapy is safe and improves symptoms, quality of life, and possibly perfusion in patients with chronic HF. PMID- 21641355 TI - Patterns of left ventricular remodeling in chronic heart failure: prevalence and prognostic implications. AB - BACKGROUND AND AIM: Many descriptors of left ventricular (LV) remodeling have important prognostic implications in patients with chronic systolic heart failure (HF). We sought to assess the prognostic value of the combination of increased LV mass with a disproportion between wall thickness and internal diameter. METHODS AND PATIENTS: Patients (n = 536) with chronic HF, ejection fraction <50% and LV end-diastolic volume index >91 mL/m(2), classified according to LV mass index and relative wall thickness (RWT), were followed up for 33 +/- 21 months. Ventricular mass was determined using a standard M-mode echocardiographic method. Relative wall thickness was defined as the ratio of (sum of interventricular septum thickness in diastole + posterior wall thickness in diastole)/LV end-diastolic diameter. RESULTS: Prevalence of the pattern of increased LV mass index, defined as LV mass index >148 g/m(2) in men and >122 g/m(2) in women, and decreased RWT (<0.34) was 29%. Multivariable predictors of all-cause mortality were age >70 years (P < .0001), New York Heart Association class >2 (P < .0001), increased LV mass index, and decreased RWT (P = .003), E wave deceleration time <=140 ms (P = .005), and male gender (P = .025). Patients with increased LV mass index and decreased RWT had a worse survival (33%) than patients with less LV mass index and normal to reduced RWT (log-rank 23.92; P < .0001). Comparisons of Cox models showed that the combination of increased mass index and decreased RWT added prognostic value to a model that included ejection fraction and end-systolic volume index. CONCLUSION: In patients with systolic HF, an independent and incremental risk of adverse outcome was associated with increased mass index and decreased RWT. PMID- 21641356 TI - Safety, feasibility, and diagnostic value of cardiac magnetic resonance imaging in patients with cardiac pacemakers and implantable cardioverters/defibrillators at 1.5 T. AB - BACKGROUND: Recent studies suggest that magnetic resonance (MR) imaging of the brain and spine may safely be performed in patients with pacemakers (PMs) and implantable cardioverter/defibrillators (ICDs), when taking adequate precautions. The aim of this study was to investigate safety, feasibility, and diagnostic value (DV) of MR imaging in cardiac applications (cardiac MR [CMR]) in patients with PMs and ICDs for the first time. METHODS: Thirty-two PM/ICD patients with a clinical need for CMR were examined. The specific absorption rate was limited to 1.5 W/kg. Devices were reprogrammed pre-CMR to minimize interference with the electromagnetic fields. Devices were interrogated pre-CMR and post-CMR and after 3 months. Troponin I levels were measured pre-CMR and post-CMR; image quality (IQ) and DV of CMR were assessed. RESULTS: All devices could be reprogrammed normally post-CMR. No significant changes of pacing capture threshold, lead impedance, and troponin I were observed. Image quality in patients with right sided devices (RSD) was better compared with that in patients with left-sided devices (LSD) (P < .05), and less myocardial segments were affected by device related artefacts (P < .05). Diagnostic value was rated as sufficiently high, allowing for diagnosis, or better in 12 (100%) of 12 patients with RSD, and only in 7 (35%) of 20 patients with LSD. CONCLUSIONS: Cardiac MR may be performed safely when limiting specific absorption rate, appropriately monitoring patients, and following device reprogramming. Cardiac MR delivers good IQ and DV in patients with RSD. Cardiac MR in patients with RSD may therefore be performed with an acceptable risk/benefit ratio, whereas the risk/benefit ratio is rather unfavorable in patients with LSD. PMID- 21641357 TI - Feasibility and accuracy of a comprehensive multidetector computed tomography acquisition for patients referred for balloon-expandable transcatheter aortic valve implantation. AB - BACKGROUND: The aim of this study was to assess the accuracy of a comprehensive multidetector computed tomography (MDCT) evaluation of the aortic annulus (AoA), coronary artery disease (CAD), and peripheral vessels in patients referred for transcatheter aortic valve implantation (TAVI). METHODS: In 60 patients referred for TAVI, the following parameters were assessed with 64-slices MDCT and compared with transesophageal echocardiography (TEE), invasive coronary angiography (ICA), and peripheral angiography: AoA maximum diameter (Max-AoA-D(MDCT)), minimum diameter (Min-AoA-D(MDCT)), and area; lumen morphology index ([Max-AoA D(MDCT)/Min-AoA-D(MDCT)]); length of the left, right, and non-coronary aortic leaflets; degree (grades 1-4) of aortic leaflet calcifications; distance between AoA and left main coronary ostium and between AoA and right coronary ostium CAD and peripheral vessel disease. RESULTS: The Max-AoA-D(MDCT) and Min-AoA-D(MDCT) were 25.1 +/- 2.8 and 21.2 +/- 2.2 mm, respectively, with high correlation versus AoA diameter measured with TEE (r = 0.82 and 0.86, respectively). The area of AoA, systolic and diastolic lumen morphology index were 410 +/- 81.5 mm(2), 1.19 +/- 0.1 and 1.22 +/- 0.11, respectively. Aortic leaflet calcification score was 3.3 +/- 0.5. The lengths of left, right, and non-coronary aortic leaflets were 14.2 +/- 2.4, 13.7.1 +/- 2.1, and 14.5 +/- 2.6 mm, whereas distances between AoA and the left main coronary ostium and between AoA, and the right coronary ostium were 13.7 +/- 2.9 and 15.8 +/- 3.5 mm, respectively. Feasibility, negative predictive value, and accuracy for CAD detection versus ICA were 87%, 100% (CI 100-100), and 96% (95% CI 94-100), respectively. All patients (N = 17) who were ineligible for TAVI were correctly detected by MDCT. CONCLUSIONS: A comprehensive MDCT evaluation of patients referred for TAVI is feasible, provides more accurate assessment than TEE of AoA morphology, and may replace peripheral angiography in all patients and ICA in patients without significant CAD. PMID- 21641358 TI - Clinical outcome and predictors for adverse events after transcatheter aortic valve implantation with the use of different devices and access routes. AB - BACKGROUND: Transcatheter aortic valve implantation (TAVI) is a treatment option for high-risk patients with severe aortic stenosis. Previous reports focused on a single device or access site, whereas little is known of the combined use of different devices and access sites as selected by the heart team. The purpose of this study is to investigate clinical outcomes of TAVI using different devices and access sites. METHODS: A consecutive cohort of 200 patients underwent TAVI with the Medtronic CoreValve Revalving system (Medtronic Core Valve LLC, Irvine, CA; n = 130) or the Edwards SAPIEN valve (Edwards Lifesciences LLC, Irvine, CA; n = 70) implanted by either the transfemoral or transapical access route. RESULTS: Device success and procedure success were 99% and 95%, respectively, without differences between devices and access site. All-cause mortality was 7.5% at 30 days, with no differences between valve types or access sites. Using multivariable analysis, low body mass index (<20 kg/m(2)) (odds ratio [OR] 6.6, 95% CI 1.5-29.5) and previous stroke (OR 4.4, 95% CI 1.2-16.8) were independent risk factors for short-term mortality. The VARC-defined combined safety end point occurred in 18% of patients and was driven by major access site complications (8.0%), life-threatening bleeding (8.5%) or severe renal failure (4.5%). Transapical access emerged as independent predictor of adverse outcome for the Valve Academic Research Consortium-combined safety end point (OR 3.3, 95% CI 1.5 7.1). CONCLUSION: A heart team-based selection of devices and access site among patients undergoing TAVI resulted in high device and procedural success. Low body mass index and history of previous stroke were independent predictors of mortality. Transapical access emerged as a risk factor for the Valve Academic Research Consortium-combined safety end point. PMID- 21641359 TI - Management and outcomes of severe aortic stenosis in cancer patients. AB - BACKGROUND: Aortic stenosis (AS) is the commonest native valve lesion, affecting 43% of all patients with valvular heart disease. The optimal treatment of severe AS in cancer patients is unknown. The purpose of this study was to assess the impact of aortic valve replacement (AVR) on survival of cancer patients with severe AS. METHODS: Cancer patients with severe AS seen at our center between January 2001 and April 2007 were identified. Baseline demographics, symptoms, cancer diagnosis, laboratory data, treatment, and outcome were collected. Patient who had AVR were matched with controls who did not have AS. RESULTS: Out of 39,071 echocardiograms performed over the study period, 1,299 had AS (3.3%), of which 50 patients (0.13%) were identified as having severe AS. Thirteen patients (27%) underwent AVR, and 35 were managed medically. Two patients underwent valvuloplasty and were excluded. Survival was significantly longer in patients with severe AS who underwent AVR and was independent of cancer status or presence of metastases. No difference in survival was found between patients who underwent AVR and matched cancer controls. In a multivariable Cox proportional hazard regression analysis, AVR was the only significant predictor of longer survival (adjusted hazard ratio = 0.22, P = .028). CONCLUSIONS: Cancer patients with severe AS who underwent AVR had an improved survival, regardless of cancer status. PMID- 21641360 TI - Effect of rosuvastatin on C-reactive protein and progression of aortic stenosis. AB - BACKGROUND: Elevated C-reactive protein (CRP) is a common finding in patients with aortic stenosis (AS) and may be associated with rapid AS progression and worse outcome. The purpose of the study was to examine the role of high sensitivity CRP and its interaction with rosuvastatin on the progression of AS. METHODS: We measured CRP at baseline, 1 year, and end of follow-up in 260 patients with a median follow-up of 3.5 years. Analyses were performed based on baseline CRP tertiles and baseline CRP >3 and <=3 mg/L. RESULTS: After adjustment for baseline characteristics, higher CRP levels were associated with age, female gender, body mass index, and lower high-density lipoprotein cholesterol levels but not with AS severity. Treatment with rosuvastatin led to a persistent decrease in CRP at 1 year and end of follow-up. Progression of AS was detected in patients in all 3 CRP tertiles, and rosuvastatin treatment had no impact on progression in all 3 tertiles. Similar findings were observed using CRP >3 mg/L as the cutpoint. Multiple linear regression showed that baseline AS velocity (P < .001), but not CRP, was the only predictor of progression of AS; age (P = .05) and baseline AS velocity (P < .001), but not CRP and rosuvastatin treatment, were predictors of outcome events. CONCLUSION: C-reactive protein does not predict severity, progression, and prognosis in patients with mild to moderate AS. Treatment with rosuvastatin reduces CRP levels but has no effect on the progression and clinical events of AS. PMID- 21641361 TI - Institutional, provider, and patient correlates of low-density lipoprotein and non-high-density lipoprotein cholesterol goal attainment according to the Adult Treatment Panel III guidelines. AB - BACKGROUND: The aim of this analysis was to identify the proportion of coronary heart disease (CHD) patients achieving guideline-recommended low-density lipoprotein cholesterol (LDL-C) and non-high-density lipoprotein cholesterol (non HDL-C) goals and to identify correlates of dual goal attainment. METHODS: We analyzed patient, provider, and facility characteristics for 21,801 CHD patients in one Veterans Affairs Hospitals Network. RESULTS: Low-density lipoprotein cholesterol goal attainment was 80%, but optional LDL-C goal attainment was 41%. Of patients with triglycerides >=200 mg/dL, 51% attained both LDL-C and non-HDL-C goals. Correlates of higher dual goal attainment included older age (65-74 years: odds ratio [OR] 1.47, 95% CI 1.28-1.69), diabetes (OR 1.33, 95% CI 1.16-1.53), obesity (OR 1.25, 95% CI 1.04-1.50), a higher number of primary care visits (OR 1.04, 95% CI 1.04-1.05), and mild increase in illness severity of patients in provider's panel (OR 1.20, 95% CI 1.0008-1.46), whereas African American patients were less likely to achieve dual lipid goals (OR 0.63, 95% CI 0.48-0.82). Receipt of care from physician (vs nonphysician) or specialist (vs primary care) provider, number of patients in provider's panel, and percentage of patients in provider's panel with diagnosis of hyperlipidemia were not associated with dual goal attainment. CONCLUSIONS: A large proportion of CHD patients attained LDL-C goal, but optional LDL-C goal attainment was low. Patients with elevated triglycerides had poor attainment of dual LDL-C and non-HDL-C goals, suggesting a treatment gap. Factors associated with dual goal attainment may identify interventions needed to improve future guideline adherence. PMID- 21641362 TI - Assessment of adiponectin and the risk of recurrent cardiovascular events in patients presenting with an acute coronary syndrome: observations from the Pravastatin Or atorVastatin Evaluation and Infection Trial-Thrombolysis in Myocardial Infarction 22 (PROVE IT-TIMI 22). AB - BACKGROUND: Adiponectin, an adipocytokine, is secreted by fatty cells and exerts a regulatory role in atherogenesis, modulating foam cell formation and cellular adhesion. In stable atherosclerosis, plasma adiponectin has been reported to be associated with both increased and decreased cardiovascular risk. Recent data have suggested a possible discordant adverse risk relationship in acute coronary syndrome (ACS). Therefore, we investigated the association between adiponectin and cardiovascular events in patients with ACS. METHODS: We measured plasma adiponectin in 3,931 patients stabilized following ACS and assessed the relationship with 2-year outcome. Patients were followed for all-cause death and major cardiovascular events. Using multivariable Cox regression, we adjusted for age, sex, race, ACS type, diabetes, smoking status, triglycerides, blood pressure, body mass index, estimated glomerular filtration rate, treatment group (atorvastatin), B-type natriuretic peptide, and C-reactive protein. RESULTS: Adiponectin correlated negatively with age, diabetes, body mass index, and triglycerides (each, P < .001) but showed a positive relationship with the risk of death (P = .01), myocardial infarction (P = .01), and heart failure (P < .001). After adjusting for clinical risk factors, B-type natriuretic peptide, and C-reactive protein, adiponectin greater than the median (4,477 ng/mL) was independently associated with an increased risk of death or myocardial infarction (hazard ratio 1.58, 95% CI 1.10-2.28, P = .013) and congestive heart failure (hazard ratio 2.17, 95% CI 1.21-3.89, P = .010). CONCLUSIONS: Higher adiponectin concentrations early after ACS are independently associated with a higher risk of recurrent cardiovascular events. This finding is directionally opposite to that observed in patients at risk for atherosclerosis and reveals the need for investigation to elucidate differences in the pathobiology of adiponectin in stable versus unstable coronary artery disease. PMID- 21641363 TI - Incremental value of echocardiographic assessment beyond clinical evaluation for prediction of death and development of heart failure after high-risk myocardial infarction. AB - BACKGROUND: Echocardiographic measurements of left ventricular (LV) function, predominantly LV ejection fraction (LVEF), have been used to define risk in patients after myocardial infarction. However, the extent to which measures of LV structure and function provide incremental prognostic value over clinical variables in survivors of high-risk myocardial infarction has not been well defined. METHODS: Predictors of death and development of heart failure were assessed in 603 patients from the Valsartan in Acute Myocardial Infarction (VALIANT) echocardiographic substudy. We used multivariable proportional hazards models to assess the individual predictive value of echocardiographic measures including left ventricular mass index, LVEF, LV volumes, left atrial volume index, right ventricular fractional area change, mitral regurgitation, and deceleration time. We adjusted for the 11 clinical variables found previously to be most associated with all-cause mortality in this cohort. Receiver operating characteristic curves obtained via binary response regression were used to assess the incremental predictive value of echocardiographic measures in predicting outcomes of death and hospital stay for heart failure. RESULTS: Each echocardiographic measure was independently associated with outcome of death or development of heart failure (all P < .002). Left ventricular ejection fraction alone added minimal prognostic value to the clinical assessment, yet adding additional echocardiographic assessments to a multivariable model improved in predicting 17-month survival free of heart failure significantly, increasing the c-statistic from 0.74 to 0.84 (P < .001). CONCLUSION: Echocardiographic measures of cardiac structure and function beyond LVEF provide important prognostic information beyond the clinical assessment. PMID- 21641364 TI - Interleukin 33 and ST2 in non-ST-elevation myocardial infarction: comparison with Global Registry of Acute Coronary Events Risk Scoring and NT-proBNP. AB - BACKGROUND: Soluble ST2 is a marker of biomechanical strain for which the natural ligand is interleukin 33 (IL-33). They have not been studied together in non-ST elevation myocardial infarction (NSTEMI). We investigated their relationship with death, heart failure (HF) readmission, and reinfarction combined (termed major adverse cardiac events [MACE]) and, separately, in unselected patients using Global Registry of Acute Coronary Events Risk Scoring (GRACE-RS) and n terminal pro B type natriuretic peptide (NT-proBNP) as benchmark comparators. METHODS: ST2 and IL-33 were measured in 577 patients 3 to 5 days after admission. Mean follow up was 532 (150-1059) days, during which 156 patients (27%) reached the primary end point. RESULTS: ST2 was higher in those who experienced MACE when compared with event-free survivors (median 782 pg/mL vs 596, P < .001), but there was no difference in IL-33 levels across any end point. Multivariate Cox regression analysis reveals that elevated ST2 is independently associated with increased risk of MACE during the long term (hazard ratio [HR] 2.01, P = .005). This relationship continues on further adjustment for either GRACE risk score or NT proBNP individually but not on adjustment for both. ST2 also independently predicts reinfarction (HR 2.48, P = .03) and 30-day mortality (HR 4.43, P = .02, c-statistic 0.73, P < .001). Adding ST2 to GRACE or to NT-proBNP did not lead to significant improvements in the c-statistic for MACE for long-term follow-up (P = .27 and P = .57, respectively) or the net reclassification index. Neither IL-33 nor its ratio with ST2 was associated with study end points. CONCLUSIONS: Elevated ST2 predicts adverse outcome in non-ST-elevation myocardial infarction but does not significantly improve risk stratification for established markers. Interleukin 33 was not related to adverse events. PMID- 21641365 TI - Long-term effects of fosinopril and pravastatin on cardiovascular events in subjects with microalbuminuria:Ten years of follow-up of Prevention of Renal and Vascular End-stage Disease Intervention Trial (PREVEND IT). AB - BACKGROUND: The PREVEND IT investigated whether treatment targeted at lowering urinary albumin excretion (UAE) would reduce adverse cardiovascular events. We obtained extended follow-up data to approximately 10 years to investigate the long-term effects of fosinopril 20 mg and pravastatin 40 mg on cardiovascular outcomes in subjects with UAE >15 mg per 24 hours. METHODS: The original PREVEND IT consisted of 864 participants and 839 survivors after 4 years. For every survivor, the primary end point determined by the combined incidence of cardiovascular mortality and hospitalization for cardiovascular morbidity was registered in several national databases and electronic hospital systems. RESULTS: Mean total follow-up of the extended PREVEND IT was 9.5 years (range 9.4 10.7 years). Four years of treatment with fosinopril was not associated with a reduction in the primary end point compared with placebo (hazard ratio 0.87, 95% CI 0.61-1.24 [P = .42]) during long-term follow-up. After 9.5 years, subjects with a baseline UAE in the upper quintile (>50 mg/24 hours) had a total event rate of 29.5% and were at a higher risk for developing cardiovascular disease compared with less UAE (hazard ratio 2.03, 95% CI 1.38-2.97 [P <= .01]). In addition, 4 years of fosinopril treatment resulted in a risk reduction of 45% (95% CI 6%-75% [P = .04]) in this group compared with placebo. Subjects originally assigned to pravastatin had no overall risk reduction in the primary end point (P = .99). CONCLUSIONS: Elevated UAE is associated with increased cardiovascular mortality and morbidity after 9.5 years of follow-up, with a doubling of the risk if the UAE is >50 mg per 24 hours. In this group, the benefits of 4-year treatment with fosinopril were sustained during posttrial follow-up for cardiovascular mortality and morbidity. We propose that UAE be used to estimate risk in the general population and that large clinical trials be designed to confirm the hypothesis that angiotensin-converting enzyme-inhibitor treatment may be beneficial in patients with mildly elevated UAE despite the absence of other comorbidities. PMID- 21641366 TI - A multinational study of the influence of health-related quality of life on in hospital outcome after coronary artery bypass graft surgery. AB - BACKGROUND: The effect of health-related quality of life on in-hospital outcomes after coronary artery bypass grafting surgery has not been investigated in international multicenter studies. We hypothesized that poor preoperative health status is associated with mortality and length of hospital stay. METHODS: In the Multicenter Study of Perioperative Ischemia Epidemiology II, preoperative Short Form 12, Mental Component Summary (MCS), and Physical Component Summary (PCS) scores were obtained prospectively from 4,811 patients (3,834 men, 977 women) undergoing coronary artery bypass grafting surgery at 72 centers in 17 countries. Primary outcome measures were in-hospital mortality and prolonged (>14 days) length of hospital stay. RESULTS: One hundred fifty-one patients (3.1%) died. After adjustment for regional differences, a 10-point reduction in MCS score was associated with higher mortality risk (odds ratio [OR] 1.17, 95% CI 1.004-1.37, P = .04) and prolonged hospital stay (OR 1.11, 95% CI 1.01-1.21, P = .03). The preoperative PCS score was not associated with mortality risk but significantly predicted prolonged length of hospital stay (OR 1.20, 95% CI 1.09-1.33, P < .001). There was no significant interaction between gender and either the MCS or the PCS score. DISCUSSION: The preoperative PCS predicted prolonged postoperative hospital stay, whereas the preoperative MCS score was an independent predictor of both prolonged length of hospital stay and mortality. Preoperative assessment of health-related quality of life factors with the Short-Form 12 might be a useful tool for risk stratification and planning for hospital discharge and rehabilitation. PMID- 21641367 TI - Depressive symptoms are related to progression of coronary calcium in midlife women: the Study of Women's Health Across the Nation (SWAN) Heart Study. AB - BACKGROUND: Major depression and depressive symptoms are associated with cardiovascular disease (CVD), but the impact of depression on early atherogenesis is less well known, particularly in women and minorities. This study examined whether depressive symptoms are associated with progression of coronary artery calcification (CAC) among women at midlife. METHODS: The SWAN is a longitudinal, multisite study assessing health and psychologic factors in midlife women. An ancillary study (SWAN Heart) evaluated subclinical atherosclerosis in women who reported no history of CVD or diabetes. In 346 women, CAC was measured twice by electron beam computed tomography, an average of 2.3 years apart. Progression, defined as an increase by >=10 Agatston units, was analyzed using relative risk (RR) regression. Depressive symptoms were assessed with the Center for Epidemiologic Studies Depression (CES-D) Scale. RESULTS: Progression of CAC was observed in 67 women (19.1%). Each 1-SD-higher CES-D score at baseline related to a 25% increased risk of CAC progression (RR 1.25, 95% CI 1.06-1.47, P = .007), adjusting for age, time between scans, ethnicity, education, menopausal status, and known CVD risk factors. This risk was similar to the risk induced by body mass index (RR 1.31, 95% CI 1.11-1.54, P = .001) and systolic blood pressure (RR 1.28, 95% CI 1.06-1.55, P = .01). CONCLUSIONS: Depressive symptoms were independently associated with progression of CAC in this cohort of midlife women. Depressive symptoms may represent a risk factor that is potentially modifiable for early prevention of CVD in women. PMID- 21641368 TI - New insights by optical coherence tomography into the differences and similarities of culprit ruptured plaque morphology in non-ST-elevation myocardial infarction and ST-elevation myocardial infarction. AB - BACKGROUND: Plaque rupture is the most common pathology associated with non-ST elevation myocardial infarction (NSTEMI) and ST-elevation myocardial infarction (STEMI). However, limited data are available regarding ruptured plaque morphology and its relationship with the clinical syndrome. This study aimed (1) to provide a morphologic description of ruptured culprit lesions by optical coherence tomography (OCT) and (2) to investigate whether ruptured plaque morphology differs between NSTEMI and STEMI. METHODS: We included 84 consecutive patients with NSTEMI and STEMI undergoing OCT study of the culprit lesion. We identified patients with plaque rupture in the OCT study and used them as the study population. Qualitative and quantitative analysis of ruptured plaque morphology was then performed, followed by a comparison of the morphological characteristics in patients with STEMI and NSTEMI. RESULTS: Fifty-five patients (70.5%) with rupture, 25 with NSTEMI, and 30 with STEMI were used for analysis. Plaque was ruptured at the minimal lumen in 34.5% of the cases, whereas 69% of the ruptures occurred at the plaque shoulder. Ruptured cap thickness was <=90 MUm in 96% of ruptured plaques. Patients with NSTEMI had greater minimal luminal area (P < .001), less lipid content (P = .01), and lower rupture length (P < .001) and length of missing fibrous cap (P < .05) compared with patients with STEMI. CONCLUSIONS: Rupture of the plaque in myocardial infarction usually occurs in sites different than the minimal lumen and at the shoulder of areas with fibrous cap measuring <=90 MUm. Patients with STEMI have greater plaque disruption and smaller minimal lumen area than patients with NSTEMI. PMID- 21641369 TI - The possibility of delayed arterial healing 5 years after implantation of sirolimus-eluting stents: serial observations by coronary angioscopy. AB - BACKGROUND: Although very late stent thrombosis occurs several years after implantation of sirolimus-eluting stent (SES), the morphologic changes of the stent beyond 2 years have not yet been systematically studied in living patients. The late vascular response to SES was therefore evaluated by serial angioscopic studies at 2 and 5 years after stent implantation. METHODS: A total of 17 patients with 17 SES underwent a repeated angioscopy procedure at 2 and 5 years. Neointimal stent coverage (NSC) was classified as follows: grade 0, presence of uncovered struts; grade 1, visible struts through a thin neointima; or grade 2, complete neointimal coverage without visible struts. For each patient, the minimum and maximum NSC grade and the existence of in-stent thrombus were recorded. RESULTS: The minimum and maximum NSC grade did not increase between the 2 and 5 years (0.59 +/- 0.51 vs 0.88 +/- 0.70, P = .17, and 1.82 +/- 0.39 vs 1.94 +/- 0.24, P = .30, respectively). The prevalence of patients with uncovered struts did not significantly decrease from 2 to 5 years (41% vs 29%, P = .49). During the follow-up period, 3 of 6 thrombi disappeared, whereas new thrombus formation was found in 3 patients without any clinical symptoms. In-stent thrombus did not decrease (35% vs 35%, P > .99). CONCLUSIONS: The current serial angioscopic study suggests that incomplete NSC and the prevalence of latent thrombus within the SES segments did not decrease from 2 to 5 years. The risk of stent thrombosis related to incomplete healing of SES may continue for an extended period. PMID- 21641370 TI - Safety and efficacy of femoral vascular closure devices in patients undergoing primary percutaneous coronary intervention for ST-elevation myocardial infarction. AB - BACKGROUND: The use of vascular closure devices (VCDs) for the reduction of access site complications following percutaneous coronary intervention (PCI) remains controversial. Patients undergoing primary PCI for acute ST-segment elevation myocardial infarction (STEMI) are at high risk of femoral artery complications. A lack of information exists regarding the use of VCDs in this group of patients because they have been routinely excluded from randomized trials. This study sought to evaluate the safety and efficacy of the routine use of VCDs after primary PCI. METHODS: A total of 558 consecutive patients undergoing primary PCI for STEMI via femoral route were studied for in-hospital outcomes through a prospective registry from January 2003 to December 2008. The primary end point was the presence of major vascular complication (MVC) defined as a composite of fatal access site bleeding, access site complication requiring interventional or surgical correction, or access site bleeding with >=3 g/dL drop in hemoglobin or requiring blood transfusion. RESULTS: Of the total patients, 464 (83.2%) received a VCD; and manual compression was used in 94 patients (16.8%). Major vascular complication occurred in 5.2% of patients. The risk of MVC was significantly lower with VCDs compared with manual compression (4.3% vs 9.6%, P = .036). Multivariable logistic regression analysis determined that VCD use remained an independent predictor of lower rate of MVC (odds ratio 0.38, 95% CI 0.17-0.91). CONCLUSIONS: The use of VCDs in patients undergoing primary PCI for STEMI is safe and is associated with lower rates of MVC compared with manual compression. PMID- 21641371 TI - Physiologic decrease of ventilatory response to exercise in the second decade of life in healthy children. AB - BACKGROUND: Cardiopulmonary exercise testing is increasingly used in children with congenital heart defects. Because of changes related to growth, the interpretation of exercise test results heavily relies on the presence of normative data. There is growing interest in the assessment of the ventilatory response to exercise in children with congenital heart disease, but normative data are lacking. METHODS: We studied 243 consecutive children (age, 13.2 +/- 2.1 years; 128 boys) with maximal cardiopulmonary exercise testing. All children had normal clinical examination and echocardiograms. In all children, the slope of the relationship between minute ventilation and carbon dioxide production (VE/VCO(2) slope) was calculated using both only data until the respiratory compensation point (VE/VCO(2RC)) and using data until peak exercise (VE/VCO(2Peak)). RESULTS: The exercise test was maximal in all children (peak respiratory exchange ratio, 1.2 +/- 0.1). For all the cohorts, VE/VCO(2Peak) slope was 28.2 +/- 3.7; and VE/VCO(2RC) slope was 24.5 +/- 3.0, whereas peak oxygen uptake was 94.6% +/- 14.0% of predicted value. Baseline spirometric function was normal in all children (vital capacity, 100% +/- 14% and forced expired volume in the first second 97% +/- 13% of predicted). From the age of 10 to 16 years, we observed a progressive decrease in both VE/VCO(2Peak) and VE/VCO(2RC) slopes (-0.833 and -0.705 per each year), with the highest reduction observed in boys. Gender-specific percentiles for both VE/VCO(2Peak) and VE/VCO(2RC) slopes were constructed. CONCLUSION: Ventilatory response to exercise expressed as VE/VCO(2) slope seems to decrease progressively in the second decade of life. Because of age-related changes, interpretation of VE/VCO(2) slopes in this age range should be based on the reported percentiles rather than on the absolute values. PMID- 21641373 TI - Potential age-dependent effects of estrogen on neural injury. AB - In 2000, approximately 10 million women were receiving hormone replacement therapy (HRT) for alleviation of menopausal symptoms. A number of prior animal studies suggested that HRT may be neuroprotective and cardioprotective. Then, in 2003, reports from the Women's Health Initiative (WHI) indicated that long-term estrogen/progestin supplementation led to increased incidence of stroke. A second branch of the WHI in women with prior hysterectomy found an even stronger correlation between estrogen supplementation alone and stroke incidence. Follow up analyses of the data, as well as data from other smaller clinical trials, have also demonstrated increased stroke severity in women receiving HRT or estrogen alone. This review examines the studies indicating that estrogen is neuroprotectant in animal models and explores potential reasons why this may not be true in postmenopausal women. Specifically, age-related differences in estrogen receptors and estrogenic actions in the brain are discussed, with the conclusion that animal models of disease must closely mimic human disease to produce clinically relevant results. PMID- 21641374 TI - Resident tissue-specific mesenchymal progenitor cells contribute to fibrogenesis in human lung allografts. AB - Fibrotic obliteration of the small airways leading to progressive airflow obstruction, termed bronchiolitis obliterans syndrome (BOS), is the major cause of poor outcomes after lung transplantation. We recently demonstrated that a donor-derived population of multipotent mesenchymal stem cells (MSCs) can be isolated from the bronchoalveolar lavage (BAL) fluid of human lung transplant recipients. Herein, we study the organ specificity of these cells and investigate the role of local mesenchymal progenitors in fibrogenesis after lung transplantation. We demonstrate that human lung allograft-derived MSCs uniquely express embryonic lung mesenchyme-associated transcription factors with a 35,000 fold higher expression of forkhead/winged helix transcription factor forkhead box (FOXF1) noted in lung compared with bone marrow MSCs. Fibrotic differentiation of MSCs isolated from normal lung allografts was noted in the presence of profibrotic mediators associated with BOS, including transforming growth factor beta and IL-13. MSCs isolated from patients with BOS demonstrated increased expression of alpha-SMA and collagen I when compared with non-BOS controls, consistent with a stable in vivo fibrotic phenotype. FOXF1 mRNA expression in the BAL cell pellet correlated with the number of MSCs in the BAL fluid, and myofibroblasts present in the fibrotic lesions expressed FOXF1 by in situ hybridization. These data suggest a key role for local tissue-specific, organ resident, mesenchymal precursors in the fibrogenic processes in human adult lungs. PMID- 21641375 TI - Skp2 is necessary for Myc-induced keratinocyte proliferation but dispensable for Myc oncogenic activity in the oral epithelium. AB - The proto-oncogene c-Myc encodes a transcription factor that is implicated in the regulation of cellular proliferation, differentiation, and apoptosis. Myc accelerates the rate of cell proliferation, at least in part, through its ability to down-regulate the expression of the cell cycle inhibitor p27(Kip1). Moreover, p27(Kip1) protein levels are regulated by ubiquitin-mediated turnover, leading to destruction by the E3 ubiquitin ligase SCF(Skp2). Therefore, we hypothesize that a lack of Skp2 expression should lead to increased p27(Kip1) levels and further inhibition of Myc-mediated proliferation and tumorigenesis. Myc expression in epithelial tissues of transgenic mice (K5-Myc) led to increased keratinocyte proliferation and the development of spontaneous tumors within the oral cavity. We generated K5-Myc-transgenic mice in an Skp2-null background. Consistent with our hypothesis, we found that Myc-mediated keratinocyte hyperproliferation was abolished by the loss of Skp2. However, Skp2 ablation did not affect Myc-driven tumorigenesis because the incidence, latency, and degree of differentiation of oral tumors were identical between K5-Myc/Skp2(+/+) and K5-Myc/Skp2(-/-) mice. Altogether, these findings suggest that Skp2 and p27(Kip1) are critical for Myc driven keratinocyte proliferation; however, Myc-mediated tumorigenesis in the oral epithelium is independent of the Skp2-p27(Kip1) axis. PMID- 21641376 TI - Chemokine receptor requirements for epidermal T-cell trafficking. AB - Inflamed skin contains CD4 T-cell subsets that express chemokine receptors CCR4, CCR6, and/or CCR10. Prior attempts to reveal the distinct role(s) of each receptor in T-cell trafficking to skin have not produced a coherent story. Different conclusions drawn by separate research groups are difficult to reconcile because of the disparate inflammation models used. Here we directly compare CD4 T cells from wild-type, CCR4(-/-), CCR6(-/-), and CCR10(-/-) mice in parallel assays of trafficking to skin. Our models require direct competition between wild-type and receptor-deficient populations for access to inflamed cutaneous sites. Major histocompatibility complex-peptide tetramers allowed us to identify antigen-specific endogenous long-term memory CD4 T cells within skin after multiple topical immunizations. We separately analyzed cells from the dermal and epidermal layers, allowing us to assess the involvement of each receptor in trafficking between dermis and epidermis. We found that CCR4 deficiency reduces accumulation of memory CD4 T cells in skin by approximately 20 fold, but neither CCR6 nor CCR10 deficiency yielded any detectable effects. Strikingly, no differences in dermal versus epidermal localization were observed for cells lacking any of these three receptors. Our findings raise the possibility that CCR6 and CCR10 play (as yet) unknown roles in cutaneous T-cell immunology, unrelated to skin-specific trafficking. PMID- 21641377 TI - Use of gene expression and pathway signatures to characterize the complexity of human melanoma. AB - A defining characteristic of most human cancers is heterogeneity, resulting from the somatic acquisition of a complex array of genetic and genomic alterations. Dissecting this heterogeneity is critical to developing an understanding of the underlying mechanisms of disease and to paving the way toward personalized treatments of the disease. We used gene expression data sets from the analysis of primary and metastatic melanomas to develop a molecular description of the heterogeneity that characterizes this disease. Unsupervised hierarchical clustering, gene set enrichment analyses, and pathway activity analyses were used to describe the genetic heterogeneity of melanomas. Patterns of gene expression that revealed two distinct classes of primary melanoma, two distinct classes of in-transit melanoma, and at least three subgroups of metastatic melanoma were identified. Expression signatures developed to predict the status of oncogenic signaling pathways were used to explore the biological basis underlying these differential patterns of expression. This analysis of activities revealed unique pathways that distinguished the primary and metastatic subgroups of melanoma. Distinct patterns of gene expression across primary, in-transit, and metastatic melanomas underline the genetic heterogeneity of this disease. This heterogeneity can be described in terms of deregulation of signaling pathways, thus increasing the knowledge of the biological features underlying individual melanomas and potentially directing therapeutic opportunities to individual patients with melanoma. PMID- 21641378 TI - Noninvasive imaging technologies reveal edema toxin as a key virulence factor in anthrax. AB - Powerful noninvasive imaging technologies enable real-time tracking of pathogen host interactions in vivo, giving access to previously elusive events. We visualized the interactions between wild-type Bacillus anthracis and its host during a spore infection through bioluminescence imaging coupled with histology. We show that edema toxin plays a central role in virulence in guinea pigs and during inhalational infection in mice. Edema toxin (ET), but not lethal toxin (LT), markedly modified the patterns of bacterial dissemination leading, to apparent direct dissemination to the spleen and provoking apoptosis of lymphoid cells. Each toxin alone provoked particular histological lesions in the spleen. When ET and LT are produced together during infection, a specific temporal pattern of lesion developed, with early lesions typical of LT, followed at a later stage by lesions typical of ET. Our study provides new insights into the complex spatial and temporal effects of B. anthracis toxins in the infected host, suggesting a greater role than previously suspected for ET in anthrax and suggesting that therapeutic targeting of ET contributes to protection. PMID- 21641379 TI - Spontaneous lung dysfunction and fibrosis in mice lacking connexin 40 and endothelial cell connexin 43. AB - Gap junction proteins (connexins) facilitate intercellular communication and serve several roles in regulation of tissue function and remodeling. To examine the physiologic effects of depleting two prominent endothelial connexins, Cx40 and Cx43, transgenic mice were generated by breeding Cx40-deficient mice (Cx40(-/ )) with a vascular endothelial cell (VEC)-specific Cx43-deficient mouse strain (VEC Cx43(-/-)) to produce double-connexin knockout mice (VEC Cx43(-/-)/Cx40(-/ )). The life span in VEC Cx43(-/-)/Cx40(-/-) mice was dramatically shortened, which correlated with severe spontaneous lung abnormalities as the mice aged including increased fibrosis, aberrant alveolar remodeling, and increased lung fibroblast content. Moreover, VEC Cx43(-/-)/Cx40(-/-) mice exhibited cardiac hypertrophy and hypertension. Because VEC Cx43(-/-)/Cx40(-/-) mice demonstrated phenotypic hallmarks that were remarkably similar to those in mice deficient in caveolin-1, pulmonary caveolin expression was examined. Lungs from VEC Cx43(-/ )/Cx40(-/-) mice demonstrated significantly decreased expression of caveolin-1 and caveolin-2. This suggests that expression of caveolin-1 may be linked to expression of Cx40 and endothelial Cx43. Moreover, the phenotype of caveolin-1(-/ ) mice and VEC Cx43(-/-)/Cx40(-/-) mice may arise via a common mechanism. PMID- 21641380 TI - Selective regulation of cardiac organic cation transporter novel type 2 (OCTN2) in dilated cardiomyopathy. AB - Organic cation transporters (OCT1-3 and OCTN1/2) facilitate cardiac uptake of endogenous compounds and numerous drugs. Genetic variants of OCTN2, for example, reduce uptake of carnitine, leading to heart failure. Whether expression and function of OCTs and OCTNs are altered by disease has not been explored in detail. We therefore studied cardiac expression, heart failure-dependent regulation, and affinity to cardiovascular drugs of these transporters. Cardiac transporter mRNA levels were OCTN2>OCT3>OCTN1>OCT1 (OCT2 was not detected). Proteins were localized in vascular structures (OCT3/OCTN2/OCTN1) and cardiomyocytes (OCT1/OCTN1). Functional studies revealed a specific drug interaction profile with pronounced inhibition of OCT1 function, for example, carvedilol [half maximal inhibitory concentration (IC50), 1.4 MUmol/L], diltiazem (IC50, 1.7 MUmol/L), or propafenone (IC50, 1.0 MUmol/L). With use of the cardiomyopathy model of coxsackievirus-infected mice, Octn2mRNA expression was significantly reduced (56% of controls, 8 days after infection). Accordingly, in endomyocardial biopsy specimens OCTN2 expression was significantly reduced in patients with dilated cardiomyopathy, whereas the expression of OCT1-3 and OCTN1 was not affected. For OCTN2 we observed a significant correlation between expression and left ventricular ejection fraction (r = 0.53, P < 0.0001) and the presence of cardiac CD3+ T cells (r = -0.45, P < 0.05), respectively. OCT1, OCT3, OCTN1, and OCTN2 are expressed in the human heart and interact with cardiovascular drugs. OCTN2 expression is selectively reduced in dilated cardiomyopathy patients and predicts the impairment of cardiac function. PMID- 21641381 TI - Direct leukocyte migration across pulmonary arterioles and venules into the perivascular interstitium of murine lungs during bleomycin injury and repair. AB - During acute lung injury and repair, leukocytes are thought to enter the lung primarily across alveolar capillaries and postcapillary venules. We hypothesized that leukocytes also migrate across pulmonary arterioles and venules, which serve as alternative sites for leukocyte influx into the lung during acute lung injury and repair. Lung sections from C57BL/6J mice up to 14 days after intratracheal bleomycin (3.33 U/kg) or saline instillation were assessed by light, fluorescence, confocal, and transmission electron microscopy for evidence of inflammatory cell sequestration and transmigration at these sites. After bleomycin treatment, large numbers of leukocytes (including neutrophils, eosinophils, and monocytes) were present in the vascular lumina and in perivascular interstitia of pulmonary arterioles and venules, as well as within the vascular walls. Leukocytes were observed within well-defined pathways in arteriolar walls and much less structured pathways in venular walls, apparently in the process of transmigration. Intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) were expressed at sites of leukocyte interaction with the luminal surface, especially in arterioles. Leukocytes appeared to exit from the vessels near collagen fibers into the perivascular interstitium. Results indicate that leukocytes can directly migrate across arteriolar and venular walls into the perivascular interstitium, which may represent an important but under-recognized pathway for leukocyte influx into the lung during injury and repair. PMID- 21641382 TI - Blockade of TSP1-dependent TGF-beta activity reduces renal injury and proteinuria in a murine model of diabetic nephropathy. AB - Transforming growth factor-beta (TGF-beta) is key in the pathogenesis of diabetic nephropathy. Thrombospondin 1 (TSP1) expression is increased in diabetes, and TSP1 regulates latent TGF-beta activation in vitro and in diabetic animal models. Herein, we investigate the effect of blockade of TSP1-dependent TGF-beta activation on progression of renal disease in a mouse model of type 1 diabetes (C57BL/6J-Ins2(Akita)) as a targeted treatment for diabetic nephropathy. Akita and control C57BL/6 mice who underwent uninephrectomy received 15 weeks of thrice weekly i.p. treatment with 3 or 30 mg/kg LSKL peptide, control SLLK peptide, or saline. The effects of systemic LSKL peptide on dermal wound healing was assessed in type 2 diabetic mice (db/db). Proteinuria (urinary albumin level and albumin/creatinine ratio) was significantly improved in Akita mice treated with 30 mg/kg LSKL peptide. LSKL treatment reduced urinary TGF-beta activity and renal phospho-Smad2/3 levels and improved markers of tubulointerstitial injury (fibronectin) and podocytes (nephrin). However, LSKL did not alter glomerulosclerosis or glomerular structure. LSKL did not increase tumor incidence or inflammation or impair diabetic wound healing. These data suggest that selective targeting of excessive TGF-beta activity through blockade of TSP1 dependent TGF-beta activation represents a therapeutic strategy for treating diabetic nephropathy that preserves the homeostatic functions of TGF-beta. PMID- 21641383 TI - Length-dependent retention of carbon nanotubes in the pleural space of mice initiates sustained inflammation and progressive fibrosis on the parietal pleura. AB - The fibrous shape of carbon nanotubes (CNTs) raises concern that they may pose an asbestos-like inhalation hazard, leading to the development of diseases, especially mesothelioma. Direct instillation of long and short CNTs into the pleural cavity, the site of mesothelioma development, produced asbestos-like length-dependent responses. The response to long CNTs and long asbestos was characterized by acute inflammation, leading to progressive fibrosis on the parietal pleura, where stomata of strictly defined size limit the egress of long, but not short, fibers. This was confirmed by demonstrating clearance of short, but not long, CNT and nickel nanowires and by visualizing the migration of short CNTs from the pleural space by single-photon emission computed tomographic imaging. Our data confirm the hypothesis that, although a proportion of all deposited particles passes through the pleura, the pathogenicity of long CNTs and other fibers arises as a result of length-dependent retention at the stomata on the parietal pleura. PMID- 21641384 TI - Inhibiting TGF-beta activity improves respiratory function in mdx mice. AB - Respiratory function is the main cause of mortality in patients with Duchenne muscular dystrophy (DMD). Elevated levels of TGF-beta play a key role in the pathophysiology of DMD. To determine whether therapeutic attenuation of TGF-beta signaling improves respiratory function, mdx mice were treated from 2 weeks of age to 2 months or 9 months of age with either 1D11 (a neutralizing antibody to all three isoforms of TGF-beta), losartan (an angiotensin receptor antagonist), or a combination of the two agents. Respiratory function was measured in nonanesthetized mice by plethysmography. The 9-month-old mdx mice had elevated Penh values and decreased breathing frequency, due primarily to decreased inspiratory flow rate. All treatments normalized Penh values and increased peak inspiratory flow, leading to decreased inspiration times and breathing frequency. Additionally, forelimb grip strength was improved after 1D11 treatment at both 2 and 9 months of age, whereas, losartan improved grip strength only at 2 months. Decreased serum creatine kinase levels (significant improvement for all groups), increased diaphragm muscle fiber density, and decreased hydroxyproline levels (significant improvement for 1D11 only) also suggested improved muscle function after treatment. For all endpoints, 1D11 was equivalent or superior to losartan; coadministration of the two agents was not superior to 1D11 alone. In conclusion, TGF-beta antagonism may be a useful therapeutic approach for treating DMD patients. PMID- 21641385 TI - Fussel-15, a new player in wound healing, is deregulated in keloid and localized scleroderma. AB - Dermal wound healing depends on highly complex interplay among various cytokines and cell types. Disruption of this process can result in impaired healing in the form of excessive scarring, as is the case in fibrotic diseases such as keloid and scleroderma. In the present study, we found Fussel-15, a new member of the Ski/Sno family of TGF-beta/BMP signaling repressors, to be expressed in early wound healing and constantly overexpressed in keloid-derived and scleroderma derived fibroblasts. Comparing the results of three-dimensional free-floating and attached-released in vitro wound healing assays, we observed that Fussel-15 is expressed during the migratory phase in the free-floating assay, indicating that Fussel-15 might play a role during fibroblast migration. Fussel-15-transfected fibroblasts showed greater migration ability in a scratch wound healing assay, compared with control-transfected cells. This migratory phenotype due to Fussel 15 was confirmed by increased peripheral F-actin localization and modifications in size, amount, and distribution of focal adhesion complexes, which were observed using F-actin and focal adhesion kinase (FAK) immunofluorescence staining, respectively. The present results suggest that expression of Fussel-15 during wound healing might promote fibroblast migration. Permanent expression of Fussel-15 in keloid and skin sclerosis fibroblasts could be involved in the pathogenesis of these conditions, but the molecular mechanism underlying this up regulation remains to be determined. PMID- 21641386 TI - beta-cell loss and beta-cell apoptosis in human type 2 diabetes are related to islet amyloid deposition. AB - Amyloid deposition and reduced beta-cell mass are pathological hallmarks of the pancreatic islet in type 2 diabetes; however, whether the extent of amyloid deposition is associated with decreased beta-cell mass is debated. We investigated the possible relationship and, for the first time, determined whether increased islet amyloid and/or decreased beta-cell area quantified on histological sections is correlated with increased beta-cell apoptosis. Formalin fixed, paraffin-embedded human pancreas sections from subjects with (n = 29) and without (n = 39) diabetes were obtained at autopsy (64 +/- 2 and 70 +/- 4 islets/subject, respectively). Amyloid and beta cells were visualized by thioflavin S and insulin immunolabeling. Apoptotic beta cells were detected by colabeling for insulin and by TUNEL. Diabetes was associated with increased amyloid deposition, decreased beta-cell area, and increased beta-cell apoptosis, as expected. There was a strong inverse correlation between beta-cell area and amyloid deposition (r = -0.42, P < 0.001). beta-Cell area was selectively reduced in individual amyloid-containing islets from diabetic subjects, compared with control subjects, but amyloid-free islets had beta-cell area equivalent to islets from control subjects. Increased amyloid deposition was associated with beta-cell apoptosis (r = 0.56, P < 0.01). Thus, islet amyloid is associated with decreased beta-cell area and increased beta-cell apoptosis, suggesting that islet amyloid deposition contributes to the decreased beta-cell mass that characterizes type 2 diabetes. PMID- 21641387 TI - Apoptosis-like cell death induction and aberrant fibroblast properties in human incisional hernia fascia. AB - Incisional hernia often occurs following laparotomy and can be a source of serious problems. Although there is evidence that a biological cause may underlie its development, the mechanistic link between the local tissue microenvironment and tissue rupture is lacking. In this study, we used matched tissue-based and in vitro primary cell culture systems to examine the possible involvement of fascia fibroblasts in incisional hernia pathogenesis. Fascia biopsies were collected at surgery from incisional hernia patients and non-incisional hernia controls. Tissue samples were analyzed by histology and immunoblotting methods. Fascia primary fibroblast cultures were assessed at morphological, ultrastructural, and functional levels. We document tissue and fibroblast loss coupled to caspase-3 activation and induction of apoptosis-like cell-death mechanisms in incisional hernia fascia. Alterations in cytoskeleton organization and solubility were also observed. Incisional hernia fibroblasts showed a consistent phenotype throughout early passages in vitro, which was characterized by significantly enhanced cell proliferation and migration, reduced adhesion, and altered cytoskeleton properties, as compared to non-incisional hernia fibroblasts. Moreover, incisional hernia fibroblasts displayed morphological and ultrastructural alterations compatible with autophagic processes or lysosomal dysfunction, together with enhanced sensitivity to proapoptotic challenges. Overall, these data suggest an ongoing complex interplay of cell death induction, aberrant fibroblast function, and tissue loss in incisional hernia fascia, which may significantly contribute to altered matrix maintenance and tissue rupture in vivo. PMID- 21641388 TI - TRPV1 involvement in inflammatory tissue fibrosis in mice. AB - We examined whether absence or blocking of transient receptor potential vanilloid subtype 1 (TRPV1) affects the level of inflammation and fibrosis/scarring during healing of injured tissue using an alkali burn model of cornea in mice. A cornea burn was produced with 1 N NaOH instilled into one eye of TRPV1-/- (KO) (n = 88) or TRPV1+/+ (n = 94) mice. Examinations of the corneal surface and eye globe size suggested that the loss of TRPV1 suppressed inflammation and fibrosis/scarring after alkali burn, and this was confirmed by histology, IHC, and gene expression analysis. The loss of TRPV1 inhibited inflammatory cell invasion and myofibroblast generation in association with reduction of expression of proinflammatory and profibrogenic components. Experiments of bone marrow transplantation between either genotype of mice showed that KO corneal tissue resident cells, but not KO bone marrow-derived cells, are responsible for KO-type wound healing with reduced inflammation and fibrosis. The absence of TRPV1 attenuated expression of transforming growth factor beta 1 (TGFbeta1) and other proinflammatory gene expression in cultured ocular fibroblasts, but did not affect TGFbeta1 expression in macrophages. Loss of TRPV1 inhibited myofibroblast transdifferentiation in cultured fibroblasts. Systemic TRPV1 antagonists reproduced the KO type of healing. In conclusion, absence or blocking of TRPV1 suppressed inflammation and fibrosis/scarring during healing of alkali-burned mouse cornea. TRPV1 is a potential drug target for improving the outcome of inflammatory/fibrogenic wound healing. PMID- 21641389 TI - Angiotensin II-induced MMP-2 activity and MMP-14 and basigin protein expression are mediated via the angiotensin II receptor type 1-mitogen-activated protein kinase 1 pathway in retinal pigment epithelium: implications for age-related macular degeneration. AB - Accumulation of various lipid-rich extracellular matrix (ECM) deposits under the retinal pigment epithelium (RPE) has been observed in eyes with age-related macular degeneration (AMD). RPE-derived matrix metalloproteinase (MMP)-2, MMP-14, and basigin (BSG) are major enzymes involved in the maintenance of ECM turnover. Hypertension (HTN) is a systemic risk factor for AMD. It has previously been reported that angiotensin II (Ang II), one of the most important hormones associated with HTN, increases MMP-2 activity and its key regulator, MMP-14, in RPE, inducing breakdown of the RPE basement membrane, which may lead to progression of sub-RPE deposits. Ang II exerts most of its actions by activating the mitogen-activated protein kinase (MAPK) signaling pathway. Herein is explored the MAPK signaling pathway as a potential key intracellular modulator of Ang II induced increase in MMP-2 activity and MMP-14 and BSG protein expression. It was observed that Ang II stimulates phosphorylation of extracellular signal-regulated kinase (ERK) and p38 MAPK in RPE cells and ERK/p38 and Jun N-terminal kinase (JNK) in mice. These effects were mediated by Ang II type 1 receptors. Blockade of ERK or p38 MAPK abrogated the increase in MMP-2 activity and MMP-14 and BSG proteins in ARPE-19 cells. A better understanding of the molecular events by which Ang II induces ECM dysregulation is of critical importance to further define its contribution to the progression of sub-RPE deposits in AMD patients with HTN. PMID- 21641390 TI - Characterization of the cysteinyl leukotriene 2 receptor in novel expression sites of the gastrointestinal tract. AB - Cysteinyl leukotrienes (cysLTs: LTC4, LTD4, and LTE4) are pro-inflammatory lipid molecules synthesized from arachidonic acid. They exert their actions on at least two cysLT receptors (CysLT1R and CysLT2R). Endothelial expression and activation of these receptors is linked to vasoactive responses and to the promotion of vascular permeability. Here we track the expression pattern of CysLT2R in a loss of-function murine model (CysLT2R-LacZ) to neurons of the myenteric and submucosal plexus in the small intestine, colonic myenteric plexus, dorsal root ganglia, and nodose ganglion. Cysteinyl leukotriene (LTC4/D4) stimulation of colonic submucosal venules elicited a greater permeability response in wild-type mice. In a dextran sulfate sodium-induced colon inflammation model, the disease activity index and colonic edema (measured by wet:dry weights and submucosal thickness) were significantly reduced in knockout (KO) mice compared to controls. Tumor necrosis factor-alpha levels in colon tissue were significantly lower in KO mice; however, myeloperoxidase activity was similar in both the KO and wild-type groups. Finally, patch-clamp recordings of basal neuronal activity of colonic projecting nociceptive neurons from dorsal root ganglia (T9-13) revealed significantly higher excitability in KO neurons compared to wild type. These results suggest that a lack of neuronal expression of CysLT2R in the murine colonic myenteric plexus attenuates colitis disease progression via a reduction in inflammation-associated tissue edema and increases neuronal sensitivity to nociceptive stimuli. PMID- 21641391 TI - Adiponectin regulation of stellate cell activation via PPARgamma-dependent and independent mechanisms. AB - In this study, we elucidated the mechanism by which adiponectin modulates hepatic stellate cell activation and fibrogenesis. Adiponectin-overexpressing transgenic mice receiving thioacetamide were resistant to fibrosis, compared with controls. In contrast, adiponectin-null animals developed severe fibrosis. Expression of collagen alpha1(I) and alpha-smooth muscle actin (alpha-SMA) mRNAs were significantly lower in adiponectin-overexpressing mice, compared with controls. In wild-type stellate cells exposed to a lentivirus encoding adiponectin, expression of peroxisome proliferator-activated receptor-gamma (PPARgamma), SREBP1c, and CEBPalpha mRNAs was significantly increased (3.2-, 4.1-, and 2.2 fold, respectively; n = 3; P < 0.05, adiponectin virus versus control), consistent with possible activation of an adipogenic transcriptional program. Troglitazone, a PPARgamma agonist, strongly suppressed up-regulation of collagen alpha1(I) and alpha-SMA mRNA in stellate cells isolated from wild-type mice; however, stellate cells from adiponectin-null animals failed to respond to troglitazone. Furthermore, in isolated stellate cells in which PPARgamma was depleted using an adenovirus-Cre-recombinase system and in which adiponectin was also overexpressed, collagen alpha1(I) and alpha-SMA were significantly inhibited. We conclude that the PPARgamma effect on stellate cell activation and the fibrogenic cascade appears to be adiponectin-dependent; however, the inhibitory effect of adiponectin on stellate cell activation was not dependent on PPARgamma, suggesting the presence of PPARgamma-dependent as well as independent pathways in stellate cells. PMID- 21641392 TI - A serrated colorectal cancer pathway predominates over the classic WNT pathway in patients with hyperplastic polyposis syndrome. AB - Hyperplastic polyposis syndrome (HPS) is characterized by the presence of multiple colorectal serrated polyps and is associated with an increased colorectal cancer (CRC) risk. The mixture of distinct precursor lesion types and malignancies in HPS provides a unique model to study the canonical pathway and a proposed serrated CRC pathway in humans. To establish which CRC pathways play a role in HPS and to obtain new support for the serrated CRC pathway, we assessed the molecular characteristics of polyps (n = 84) and CRCs (n = 19) in 17 patients with HPS versus control groups of various sporadic polyps (n = 59) and sporadic microsatellite-stable CRCs (n = 16). In HPS and sporadic polyps, APC mutations were exclusively identified in adenomas, whereas BRAF mutations were confined to serrated polyps. Six of 19 HPS CRCs (32%) were identified in a serrated polyp. Mutation analysis performed in the CRC and the serrated component of these lesions showed identical BRAF mutations. One HPS CRC was located in an adenoma, both components harboring an identical APC mutation. Overall, 10 of 19 HPS CRCs (53%) carried a BRAF mutation versus none in control group CRCs (P = 0.001). Six BRAF-mutated HPS CRCs (60%) were microsatellite unstable owing to MLH1 methylation. These findings provide novel supporting evidence for the existence of a predominant serrated CRC pathway in HPS, generating microsatellite-stable and microsatellite-instable CRCs. PMID- 21641393 TI - Changes in autophagic response in patients with chronic hepatitis C virus infection. AB - Autophagy is a regulated process that can be involved in the elimination of intracellular microorganisms and in antigen presentation. Some in vitro studies have shown an altered autophagic response in hepatitis C virus infected hepatocytes. The present study aimed at evaluating the autophagic process in the liver of chronic hepatitis C (CHC) patients. Fifty-six CHC patients and 47 control patients (8 with nonalcoholic steatohepatitis or alcoholic liver disease, 18 with chronic heptatitis B virus infection, and 21 with no or mild liver abnormalities at histological examination) were included. Autophagy was assessed by means of electron microscopy and microtubule-associated protein light chain 3 immunoblotting. Using light chain 3 immunoblotting, the form present on autophagic vesicle (light chain 3-II) was significantly higher in CHC patients than in controls (P < 0.05). Using quantitative electron microscopy analysis, the median number of autophagic vesicles observed in hepatocytes from CHC patients was sixfold higher than in overall controls (P < 0.001). In contrast, there was no difference between CHC patients and controls in the number of mature lysosomes with electron-dense contents arguing in favor of a lack of fusion between autophagosome and lysosome. Neither genotype nor viral load influenced the autophagy level. In conclusion, autophagy is altered in hepatocytes from CHC patients, likely due to a blockade of the last step of the autophagic process. PMID- 21641394 TI - Autoreactive cytotoxic T lymphocytes acquire higher expression of cytotoxic effector markers in the islets of NOD mice after priming in pancreatic lymph nodes. AB - Cytotoxic T lymphocytes (CTLs) that cause type 1 diabetes are activated in draining lymph nodes and become concentrated as fully active CTLs in inflamed pancreatic islets. It is unclear whether CTL function is driven by signals received in the lymph node or also in the inflamed tissue. We studied whether the development of cytotoxicity requires further activation in islets. Autoreactive CTLs found in the islets of diabetes-prone NOD mice had acquired much higher expression of the cytotoxic effector markers granzyme B, interferon gamma, and CD107a than had those in the pancreatic lymph node (PLN). Increased expression seemed to result from stimulation in the islet itself. T cells held up from migrating from the PLN by administration of the sphingosine-1-phosphate agonist FTY720 did not increase expression of cytotoxic molecules in the PLN. Stimulation did not require antigen presentation or cytokine secretion by the target beta cells because it was not affected by the absence of class I major histocompatibility complex expression or by the overexpression of suppressor of cytokine signaling-1. Activation of CD40-expressing cells stimulated increased CTL function and beta-cell destruction, suggesting that signals derived from CD40 expressing cells promote the acquisition of cytotoxicity in the islet environment. These data provide in vivo evidence that stimulation of cytotoxic effector molecule expression occurs in inflamed islets and is independent of beta cells. PMID- 21641395 TI - Borrelia burgdorferi RST1 (OspC type A) genotype is associated with greater inflammation and more severe Lyme disease. AB - Evidence is emerging for differential pathogenicity among Borrelia burgdorferi genotypes in the United States. By using two linked genotyping systems, ribosomal RNA intergenic spacer type (RST) and outer surface protein C (OspC), we studied the inflammatory potential of B. burgdorferi genotypes in cells and patients with erythema migrans or Lyme arthritis. When macrophages were stimulated with 10 isolates of each RST1, RST2, or RST3 strain, RST1 (OspC type A)-stimulated cells expressed significantly higher levels of IL-6, IL-8, chemokine ligand (CCL) 3, CCL4, tumor necrosis factor, and IL-1beta, factors associated with innate immune responses. In peripheral blood mononuclear cells, RST1 strains again stimulated significantly higher levels of these mediators. Moreover, compared with RST2, RST1 isolates induced significantly more interferon (IFN)-alpha, IFN-gamma, and CXCL10, which are needed for adaptive immune responses; however, OspC type I (RST3) approached RST1 (OspC type A) in stimulating these adaptive immune mediators. Similarly, serum samples from patients with erythema migrans who were infected with the RST1 genotype had significantly higher levels of almost all of these mediators, including exceptionally high levels of IFN-gamma-inducible chemokines, CCL2, CXCL9, and CXCL10; and this pronounced inflammatory response was associated with more symptomatic infection. Differences among genotypes were not as great in patients with Lyme arthritis, but those infected with RST1 strains more often had antibiotic-refractory arthritis. Thus, the B. burgdorferi RST1 (OspC type A) genotype, followed by the RST3 (OspC type I) genotype, causes greater inflammation and more severe disease, establishing a link between spirochetal virulence and host inflammation. PMID- 21641396 TI - Aberrant CD8+ T-cell responses and memory differentiation upon viral infection of an ataxia-telangiectasia mouse model driven by hyper-activated Akt and mTORC1 signaling. AB - Immune system-related pathology is common in ataxia-telangiectasia (A-T) patients and mice that lack the protein kinase, A-T mutated (ATM). However, it has not been studied how ATM influences immune responses to a viral infection. Using the lymphocytic choriomeningitis virus (LCMV) infection model, we show that ATM(-/-) mice, despite having fewer naive CD8+ T cells, effectively clear the virus. However, aberrant CD8+ T-cell responses are observed, including defective expansion and contraction, effector-to-memory differentiation, and a switch in viral-epitope immunodominance. T-cell receptor-activated, but not naive, ATM(-/-) splenic CD8+ T cells have increased ribosomal protein S6 and Akt phosphorylation and do not proliferate well in response to IL-15, a cytokine important for memory T-cell development. Accordingly, pharmacological Akt or mammalian target of rapamycin complex 1 (mTORC1) inhibition during T-cell receptor activation alone rescues the IL-15 proliferation defect. Finally, rapamycin treatment during LCMV infection in vivo increases the number of memory T cells in ATM(-/-) mice. Altogether, these results show that CD8+T cells lacking ATM have hyperactive Akt and mTORC1 signaling in response to T-cell receptor activation, which results in aberrant cytokine responses and memory T-cell development. We speculate that similar signaling defects contribute to the immune system pathology of A-T, and that inhibition of Akt and/or mTORC1 may be of therapeutic value. PMID- 21641397 TI - Extracellular superoxide dismutase in macrophages augments bacterial killing by promoting phagocytosis. AB - Extracellular superoxide dismutase (EC-SOD) is abundant in the lung and limits inflammation and injury in response to many pulmonary insults. To test the hypothesis that EC-SOD has an important role in bacterial infections, wild-type and EC-SOD knockout (KO) mice were infected with Escherichia coli to induce pneumonia. Although mice in the EC-SOD KO group demonstrated greater pulmonary inflammation than did wild-type mice, there was less clearance of bacteria from their lungs after infection. Macrophages and neutrophils express EC-SOD; however, its function and subcellular localization in these inflammatory cells is unclear. In the present study, immunogold electron microscopy revealed EC-SOD in membrane bound vesicles of phagocytes. These findings suggest that inflammatory cell EC SOD may have a role in antibacterial defense. To test this hypothesis, phagocytes from wild-type and EC-SOD KO mice were evaluated. Although macrophages lacking EC SOD produced more reactive oxygen species than did cells expressing EC-SOD after stimulation, they demonstrated significantly impaired phagocytosis and killing of bacteria. Overall, this suggests that EC-SOD facilitates clearance of bacteria and limits inflammation in response to infection by promoting bacterial phagocytosis. PMID- 21641398 TI - Human leukocyte antigen class II transgenic mouse model unmasks the significant extrahepatic pathology in toxic shock syndrome. AB - Among the exotoxins produced by Staphylococcus aureus and Streptococcus pyogenes, the superantigens (SAgs) are the most potent T-cell activators known to date. SAgs are implicated in several serious diseases including toxic shock syndrome (TSS), Kawasaki disease, and sepsis. However, the immunopathogenesis of TSS and other diseases involving SAgs are still not completely understood. The commonly used conventional laboratory mouse strains do not respond robustly to SAgs in vivo. Therefore, they must be artificially rendered susceptible to TSS by using sensitizing agents such as d-galactosamine (d-galN), which skews the disease exclusively to the liver and, hence, is not representative of the disease in humans. SAg-induced TSS was characterized using transgenic mice expressing HLA class II molecules that are extremely susceptible to TSS without d-galN. HLA-DR3 transgenic mice recapitulated TSS in humans with extensive multiple-organ inflammation affecting the lung, liver, kidneys, heart, and small intestines. Heavy infiltration with T lymphocytes (both CD4(+) and CD8+), neutrophils, and macrophages was noted. In particular, the pathologic changes in the small intestines were extensive and accompanied by significantly altered absorptive functions of the enterocytes. In contrast to massive liver failure alone in the d galN sensitization model of TSS, findings of the present study suggest that gut dysfunction might be a key pathogenic event that leads to high morbidity and mortality in humans with TSS. PMID- 21641399 TI - Role of p47phox in antigen-presenting cell-mediated regulation of humoral immunity in mice. AB - Microbial-induced inflammation is important for eliciting humoral immunity. Genetic defects of NADPH oxidase 2-based proteins interrupt phagocyte superoxide generation and are the basis for the human immunodeficiency chronic granulomatous disease (CGD). Hyperinflammation is also a significant clinical manifestation of CGD. Herein, we evaluated humoral immunity in the phagocyte oxidase p47(phox) deficient model of CGD and found that UV-inactivated Streptococcus pneumoniae and Listeria monocytogenes (Lm) elicited higher specific antibody (Ab) titers in p47(phox-/-) mice than wild-type (WT) mice. Both organisms elicited robust and distinct antigen-presenting cell maturation phenotypes, including IL-12 hypersecretion, and higher major histocompatibility complex II and costimulatory protein expression in Lm-stimulated p47(phox-/-) dendritic cells (DCs) relative to WT DCs. Furthermore, p47(phox-/-) DCs pulsed with Lm and adoptively transferred into naive WT mice elicited Ab titers, whereas Lm-pulsed WT DCs did not elicit these titers. The observed robust p47(phox-/-) mouse humoral response was recapitulated with live Lm and sustained in vivo in p47(phox-/-) mice. Notably, anti-serum samples from p47(phox-/-) mice that survived secondary Lm infection were protective in WT and p47(phox-/-) mice that were rechallenged with secondary lethal Lm infection. These findings demonstrate a novel benefit of NADPH oxidase 2 deficiency (ie, dependent inflammation in antigen-presenting cell mediated humoral immunity) and that anti-Lm Ab can be protective in an immunodeficient CGD host. PMID- 21641400 TI - Systemic low-dose UVB inhibits CD8 T cells and skin inflammation by alternative and novel mechanisms. AB - Exposure to UVB radiation before antigen delivery at an unirradiated site inhibits functional immunological responses. Mice treated dorsally with suberythemal low-dose UVB and immunized with ova in abdominal skin generated ova specific CD8 T cells with a significantly decreased activation, expansion, and cytotoxic activity compared with unirradiated mice. UVB also impaired the delayed type hypersensitivity (DTH) reaction to ova. Transfer of CD4+CD25+cells from UVB exposed mice did not suppress the ova-specific CD8 T-cell response or DTH reaction in unexposed mice, confirming that systemic low-dose UVB does not induce long-lived functional regulatory CD4+CD25+ T cells. Repairing cyclobutane pyrimidine dimer-type DNA damage and blocking aryl hydrocarbon receptor signaling also did not reverse the immunosuppressive effect of UVB on ova-specific CD8 T cells and DTH, suggesting that cyclobutane pyrimidine dimers and the aryl hydrocarbon receptor are not required in systemic low-dose UVB-induced immunosuppression. The known UVB chromophore, cis-urocanic acid, and reactive oxygen species triggered the inhibition of DTH caused by UVB, but they were not involved in the modulation of CD8 T cells. These findings indicate that systemic low-dose UVB impedes the primary response of antigen-specific CD8 T cells by a novel mechanism that is independent of pathways known to be involved in systemic suppression of DTH. PMID- 21641401 TI - Effect of macrophage migration inhibitory factor (MIF) in human placental explants infected with Toxoplasma gondii depends on gestational age. AB - Because macrophage migration inhibitory factor (MIF) is a key cytokine in pregnancy and has a role in inflammatory response and pathogen defense, the objective of the present study was to investigate the effects of MIF in first- and third-trimester human placental explants infected with Toxoplasma gondii. Explants were treated with recombinant MIF, IL-12, interferon-gamma, transforming growth factor-beta1, or IL-10, followed by infection with T. gondii RH strain tachyzoites. Supernatants of cultured explants were assessed for MIF production. Explants were processed for morphologic analysis, immunohistochemistry, and real time PCR analysis. Comparison of infected and stimulated explants versus noninfected control explants demonstrated a significant increase in MIF release in first-trimester but not third-trimester explants. Tissue parasitism was higher in third- than in first-trimester explants. Moreover, T. gondii DNA content was lower in first-trimester explants treated with MIF compared with untreated explants. However, in third-trimester explants, MIF stimulus decreased T. gondii DNA content only at the highest concentration of the cytokine. In addition, high expression of MIF receptor was observed in first-trimester placental explants, whereas MIF receptor expression was low in third-trimester explants. In conclusion, MIF was up-regulated and demonstrated to be important for control of T. gondii infection in first-trimester explants, whereas lack of MIF up regulation in third-trimester placentas may be involved in higher susceptibility to infection at this gestational age. PMID- 21641402 TI - Placental TNF-alpha signaling in illness-induced complications of pregnancy. AB - Maternal infections are implicated in a variety of complications during pregnancy, including pregnancy loss, prematurity, and increased risk of neurodevelopmental disorders in the child. Here, we show in mice that even mild innate immune activation by low-dose lipopolysaccharide in early pregnancy causes hemorrhages in the placenta and increases the risk of pregnancy loss. Surviving fetuses exhibit hypoxia in the brain and impaired fetal neurogenesis. Maternal Toll-like receptor 4 signaling is a critical mediator of this process, and its activation is accompanied by elevated proinflammatory cytokines in the placenta. We evaluated the role of tumor necrosis factor-alpha (TNF-alpha) signaling and show that TNF receptor 1 (TNFR1) is necessary for the illness-induced placental pathology, accompanying fetal hypoxia, and neuroproliferative defects in the fetal brain. We also show that placental TNFR1 in the absence of maternal TNFR1 is sufficient for placental pathology to develop and that a clinically relevant TNF-alpha antagonist prevents placental pathology and fetal loss. Our observations suggest that the placenta is highly sensitive to proinflammatory signaling in early pregnancy and that TNF-alpha is an effective target for preventing illness-related placental defects and related risks to the fetus and fetal brain development. PMID- 21641403 TI - Neuroprotective effects of the cellular prion protein in autoimmune optic neuritis. AB - Although the pathologic role of the prion protein in transmissible spongiform encephalopathic diseases has been widely investigated, the physiologic role of the cellular prion protein (PrP(C)) is not known. Among the many functions attributed to PrP(C), there is increasing evidence that it is involved in cell survival and mediates neuroprotection. A potential role in the immune response has also been suggested. However, how these two functions interplay in autoimmune disease is unclear. To address this, autoimmune optic neuritis, a model of multiple sclerosis, was induced in C57Bl/6 mice, and up-regulation of PrP(C) was observed throughout the disease course. In addition, compared with wild-type mice, in PrP(C)-deficient mice and mice overexpressing PrP(C), histopathologic analysis demonstrated that optic neuritis was exacerbated, as indicated by axonal degeneration, inflammatory infiltration, and demyelination. However, significant neuroprotection of retinal ganglion cells, the axons of which form the optic nerve, was observed in mice that overexpressed PrP(C). Conversely, mice lacking PrP(C) demonstrated significantly more neurodegeneration. This suggests that PrP(C) may have a neuroprotective function independent of its role in regulating the immune response. PMID- 21641404 TI - Identification of cells with colony-forming activity, self-renewal capacity, and multipotency in ovarian endometriosis. AB - Endometriosis, the growth of endometrial tissue outside the uterine cavity, is a common gynecological disorder affecting 10% to 15% of women in their reproductive years. Retrograde menstrual shedding containing endometrial stem/progenitor cells has been postulated to be involved in its pathogenesis. In this study, we identified putative endometriotic stem/progenitor cells by their colony-forming potential, self-renewal capacity, and multipotency. Purified epithelial and stromal cells isolated from ovarian endometriotic cysts formed large and small colony-forming units (CFUs) in clonogenic assay. The colony-forming activity of epithelial and stromal cells was found to differ greatly between autologous endometrium and ovarian endometrioma samples. The large CFUs could propagate more than the small CFUs. The endometriotic epithelial small CFUs expressed epithelial markers (epithelial cell adhesion molecule, cytokeratin, and alpha6 integrin); only occasional large CFUs expressed alpha6 integrin. Aside from the expression of fibroblast markers, stromal CFUs also expressed three somatic stem cell markers: sal-like 4, CD133, and Musashi-1. Endometriotic stromal cells derived from large CFUs could differentiate into four mesenchymal lineages when cultured in the respective inducing-media, as determined by histochemical staining and RT PCR of lineage specific markers. These findings demonstrate that ovarian endometrioma contains a subset of cells displaying somatic stem cell properties. PMID- 21641405 TI - In vivo invasion of head and neck squamous cell carcinoma cells does not require macrophages. AB - Invasion of tumor cells into the local stroma is an important component in cancer progression. Here we report studies of the in vivo invasion of head and neck squamous cell carcinoma (HNSCC) cells in response to applied gradients of a growth factor [epidermal growth factor (EGF)] and a chemokine (CXCL12), using orthotopic floor-of-mouth models. Analysis of the invading cells indicated that >75% of them were tumor cells, about 15% macrophages, and <10% were unidentified. Surprisingly, although macrophages invaded together with tumor cells, macrophage contributions were not required for HNSCC invasion. CXCL12-induced in vivo invasion of HNSCC cells was also observed and found to occur via a unidirectional transactivation of epidermal growth factor receptor (EGFR) through CXCR4. Inhibition of tumor necrosis factor-alpha-converting enzyme using TNF-alpha protease inhibitor-2 selectively inhibited CXCL12-induced invasion but not EGF induced invasion, consistent with CXCL12 activation of EGFR via release of EGFR ligands. PMID- 21641406 TI - Secretory leukocyte protease inhibitor (SLPI) expression and tumor invasion in oral squamous cell carcinoma. AB - Differential expression of secretory leukocyte protease inhibitor (SLPI) impacts on tumor progression. SLPI directly inhibits elastase and other serine proteases, and regulates matrix metalloproteinases, plasminogen activation, and plasmin downstream targets to influence invasion. We examined tissues from human oral squamous cell carcinoma (OSCC) for SLPI expression in parallel with proteases associated with tumor progression and evaluated their relationships using tumor cell lines. Significantly decreased SLPI was detected in OSCC compared to normal oral epithelium. Furthermore, an inverse correlation between SLPI and histological parameters associated with tumor progression, including stage of invasion, pattern of invasion, invasive cell grade, and composite histological tumor score was evident. Conversely, elevated plasmin and elastase were positively correlated with histological parameters of tumor invasion. In addition to its known inhibition of elastase, we identify SLPI as a novel inhibitor of plasminogen activation through its interaction with annexin A2 with concomitant reduced plasmin generation by macrophages and OSCC cell lines. In an in vitro assay measuring invasive activity, SLPI blocked protease-dependent tumor cell migration. Our data suggest that SLPI may possess antitumorigenic activity by virtue of its ability to interfere with multiple requisite proteolytic steps underlying tumor cell invasion and may provide insight into potential stratification of oral cancer according to risk of occult metastasis, guiding treatment strategies. PMID- 21641407 TI - Glyceryl trinitrate inhibits hypoxia-induced release of soluble fms-like tyrosine kinase-1 and endoglin from placental tissues. AB - Preeclampsia is associated with increased circulating levels of proinflammatory molecules, such as soluble fms-like tyrosine kinase 1 (sFlt-1) and soluble endoglin (sEng). On release by an inadequately perfused placenta into the maternal circulation, these molecules cause systemic endothelial dysfunction and the associated hypertension and proteinuria that characterize preeclampsia. We previously showed that glyceryl trinitrate (GTN) inhibits hypoxia/reoxygenation induced apoptosis in the syncytiotrophoblast of term chorionic villi explants. Herein, we demonstrate that GTN inhibits the release of sFlt-1 and sEng from term chorionic villi explants exposed to hypoxia. Although transcript levels and secretion of sFlt-1 and sEng increased in explants exposed to hypoxia, low concentrations of GTN significantly inhibited the hypoxia-induced expression of these molecules at the mRNA and protein levels. Treatment of explants with GTN also prevented the hypoxia-induced accumulation of hypoxia-inducible factor 1alpha, a key mediator of cellular adaptations to hypoxia. Furthermore, knockdown of hypoxia-inducible factor-1alpha inhibited the hypoxia-induced secretion of sFlt-1 and sEng. This study provides evidence that hypoxia induces the release of sFlt-1 and sEng in the placenta via a mechanism that is inhibited by low concentrations of GTN. Our findings indicate that GTN may have potential applications in the treatment and/or prevention of preeclampsia. PMID- 21641408 TI - Mouse vein graft hemodynamic manipulations to enhance experimental utility. AB - Mouse models serve as a tool to study vein graft failure. However, in wild-type mice, there is limited intimal hyperplasia, hampering efforts to identify anti intimal hyperplasia therapies. Furthermore, vein graft wall remodeling has not been well quantified in mice. We hypothesized that simple hemodynamic manipulations can reproducibly augment intimal hyperplasia and remodeling end points in mouse vein grafts, thereby enhancing their experimental utility. Mouse inferior vena cava-to-carotid interposition isografts were completed using an anastomotic cuff technique. Three flow restriction manipulations were executed by ligating outflow carotid branches, creating an outflow common carotid stenosis, and constructing a midgraft stenosis. Flowmetry and ultrasonography were used perioperatively and at day 28. All ligation strategies decreased the graft flow rate and wall shear stress. Morphometry showed that intimal thickness increased by 26% via carotid branch ligation and by 80% via common carotid stenosis. Despite similar mean flow rates and shear stresses among the three manipulations, the flow waveform amplitudes were lowest with common carotid stenosis. The disordered flow of the midgraft stenosis yielded poststenotic dilatation. The creation of an outflow common carotid stenosis generates clinically relevant (poor runoff) vein graft low wall shear stress and offers a technically flexible method for enhancing the intimal hyperplasia response. Midgraft stenosis exhibits poststenotic positive wall remodeling. These reproducible approaches offer novel strategies for increasing the utility of mouse vein graft models. PMID- 21641409 TI - Increased vascular delivery and efficacy of chemotherapy after inhibition of platelet-derived growth factor-B. AB - Inhibition of platelet-derived growth factor-B (PDGF-B) has multiple effects on tumors, including loss of pericytes, regression of some vessels, normalization of other vessels, and reduction of interstitial pressure. PDGF-B inhibition also increases the efficacy of cancer therapeutics, but the role on tumor vessel efficiency and drug delivery is unclear. We sought to determine whether inhibition of PDGF-B signaling can increase delivery and efficacy of cyclophosphamide in Lewis lung carcinomas or RIP-Tag2 tumors. PDGF-B blockade in Lewis lung carcinoma tumors by the DNA aptamer AX102 for 14 days increased the number of perfused tumor vessels marked by lectin in the bloodstream by 50%. AX102 also increased the width of sleeves of viable tumor cells around blood vessels by 66%, increased tumor cell proliferation by 90%, and increased intratumoral delivery of Hoechst 33342 by 78%. A low dose of cyclophosphamide (20 mg/kg) reduced tumor cell proliferation by 31% when combined with AX102 but not when given alone. Synergy of cyclophosphamide and AX102 on tumor cell proliferation also was found in RIP-Tag2 tumors. Similarly, the PDGF receptor signaling inhibitor imatinib increased delivery of cyclophosphamide and reduced tumor burden in RIP-Tag2 mice, without evidence of tumor cell sensitization to chemotherapy. Together, these findings indicate that inhibition of PDGF-B signaling promotes the delivery and efficacy of chemotherapeutic agents by increasing the efficiency of tumor blood vessels. PMID- 21641411 TI - Nanoparticle-assisted MALDI-TOF MS combined with seed-layer surface preparation for quantification of small molecules. AB - Despite the advantages of simplicity and high-throughput detection that matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has over other methods, quantitative analysis of low-molecular-weight analyte is hampered by interference from matrix-derived background noise and signal fluctuation due to the inhomogeneous MALDI sample surface. Taking advantage of improved sample homogeneity through matrix-conjugated magnetic nanoparticles (matrix@MNP) and the seed-layer method, we report a new strategy for the rapid identification and quantification of drugs in urine samples, using morphine and 7 aminoflunitrazepam (7-aminoFM2) as model compounds. To our knowledge, this is the first attempt using the seed-layer method for small molecule analysis. By applying the proposed seed-layer method, which was specifically optimized for the 2,5-dihydroxybenzoic acid@MNP (DHB@MNP) matrix, homogeneous sample crystallization examined by microscopy analysis was obtained that generated reproducible MALDI signals (RSD<10.0%). For urine sample analysis, simple liquid liquid extraction as a sample pretreatment step effectively reduced the ion suppression effect caused by the endogenous components in urine; good recoveries (82-90%) were obtained with a small ion suppression effect (<14% of signal decrease). This newly developed method demonstrated good quantitation linearity over a range of 50-2000 ng mL(-1) (R(2)>0.996) with reduced signal variation (RSD<10.0%). The detection limit is 30 ng mL(-1) with good precision (intra-day, 2.0-9.3%; inter-day, 5.0-10.0%) and accuracy (intra-day, 95.0-106.0%; inter-day, 103.0-115.5%). The nanoparticle-assisted MALDI-TOF MS combined with seed-layer surface preparation provides a rapid, efficient and accurate platform for the quantification of small molecules in urine samples. PMID- 21641410 TI - Redistribution and hemostatic action of recombinant activated factor VII associated with platelets. AB - Clinical evidence accumulated from hemophilic patients during prophylaxis with recombinant activated factor VII (rFVIIa) suggests that the duration of the hemostatic action of rFVIIa exceeds its predicted plasma half-life. Mechanisms involved in this outcome have not been elucidated. We have investigated in vitro the redistribution of rFVIIa in platelets from healthy donors, patients with FVII deficiency, and one patient with Bernard-Soulier syndrome. Platelet-rich plasma was exposed to rFVIIa (3 to 60 MUg/mL). Flow cytometry, immunocytochemistry, and coagulation tests were applied to detect and quantify rFVIIa. The hemostatic effect of rFVIIa associated to platelets was evaluated using perfusion models. Our studies revealed a dose-dependent association of rFVIIa to the platelet cytoplasm with redistribution into the open canalicular system, and alpha granules. Mechanisms implicated in the internalization are multiple, involve GPIb and GPIV, and require phospholipids and cytoskeletal assembly. After platelet activation with thrombin, platelets exposed rFVIIa on their membrane. Perfusion studies revealed that the presence of 30% of platelets containing FVIIa improved platelet aggregate formation and enhanced fibrin generation (P < 0.01 versus control). Our results indicate that, at therapeutic concentrations, rFVIIa can be internalized into platelets, where it is protected from physiological clearance mechanisms and can still promote hemostatic activity. Redistribution of rFVIIa into platelets may explain the prolonged prophylactic effectiveness of rFVIIa in hemophilia. PMID- 21641412 TI - Automated optimization and construction of chemometric models based on highly variable raw chromatographic data. AB - Direct chemometric interpretation of raw chromatographic data (as opposed to integrated peak tables) has been shown to be advantageous in many circumstances. However, this approach presents two significant challenges: data alignment and feature selection. In order to interpret the data, the time axes must be precisely aligned so that the signal from each analyte is recorded at the same coordinates in the data matrix for each and every analyzed sample. Several alignment approaches exist in the literature and they work well when the samples being aligned are reasonably similar. In cases where the background matrix for a series of samples to be modeled is highly variable, the performance of these approaches suffers. Considering the challenge of feature selection, when the raw data are used each signal at each time is viewed as an individual, independent variable; with the data rates of modern chromatographic systems, this generates hundreds of thousands of candidate variables, or tens of millions of candidate variables if multivariate detectors such as mass spectrometers are utilized. Consequently, an automated approach to identify and select appropriate variables for inclusion in a model is desirable. In this research we present an alignment approach that relies on a series of deuterated alkanes which act as retention anchors for an alignment signal, and couple this with an automated feature selection routine based on our novel cluster resolution metric for the construction of a chemometric model. The model system that we use to demonstrate these approaches is a series of simulated arson debris samples analyzed by passive headspace extraction, GC-MS, and interpreted using partial least squares discriminant analysis (PLS-DA). PMID- 21641413 TI - Sensitive electrochemical immunoassay of carcinoembryonic antigen with signal dual-amplification using glucose oxidase and an artificial catalase. AB - A new dual-amplification strategy of electrochemical signal based on the catalytic recycling of the product was developed for the antigen-antibody interaction by glucose oxidase (GOD)- conjugated gold-silver hollow microspheres (AuAgHSs) coupled with an artificial catalase, Prussian blue nanoparticles (PB), on a graphene-based immunosensing platform. The first signal amplification introduced in this study was based on the labeled GOD on the AuAgHSs toward the catalytic oxidation of glucose. The generated H(2)O(2) was catalytically reduced by the immobilized PB on the graphene nanosheets with the second amplification. With a sandwich-type immunoassay format, carcinoembryonic antigen (CEA) was monitored as a model analyte by using the synthesized AuAgHSs as labels in pH 6.0 phosphate buffer containing 10mM glucose. Under optimal conditions, the electrochemical immunosensor exhibited a wide dynamic range of 0.005-50 ng mL(-1) with a low detection limit (LOD) of 1.0 pg mL(-1) CEA (at 3sigma). Both the intra and inter-assay coefficients of variation (CVs) were lower than 10%. The specificity and stability of the immunosensor were acceptable. In addition, the assay was evaluated for clinical serum specimens, and received a good correlation with those obtained by the referenced electrochemiluminescent (ECL). PMID- 21641414 TI - Spectroelectrochemical determination of the electron consumption. AB - Measurements of UV-vis spectra as a function of the consumed charge in an optically transparent thin layer cell (OTTLE) offer a fast, small-volume and reliable method for the determination of the number of electrons n exchanged in the redox reaction. The determination of n is based on the calibration procedure using the ferrocene oxidation and the methyl viologen reduction. Virtues of this method are demonstrated on the determination of n of two oligomers of 'extended viologens' with multiple redox centers and correspondingly large n. The described method eliminates problems of side reactions of reactive intermediates with the parent oxidized form, which often impede a reliable estimation of n by the established coulometric method. PMID- 21641415 TI - Direct electrochemistry of horseradish peroxidase immobilized on electrografted 4 ethynylphenyl film via click chemistry. AB - In this paper, a simple two-step approach for redox protein immobilization was introduced. Firstly, alkynyl-terminated film was formed on electrode surface by electrochemical reduction of 4-ethylnylphenyl (4-EP) diazonium compound. Then, horseradish peroxidase (HRP) modified with azido group was covalently immobilized onto the electrografted film via click reaction. Reflection absorption infrared (RAIR) spectroscopy and electrochemical methods were used to characterize the modification process. The results indicate that HRP retains its native structure and shows fast direct electron transfer. Moreover, the immobilized HRP shows excellent electrocatalytic reduction activity toward H(2)O(2) with a linear range of 5.0*10(-6) to 9.3*10(-4) mol L(-1). PMID- 21641416 TI - Extraction and analysis of avermectines in agricultural soils by microwave assisted extraction and ultra high performance liquid chromatography coupled to tandem mass spectrometry. AB - A method for the analysis of avermectines (abamectine, doramectine and ivermectine) in soils has been developed. The analytes are extracted with acetonitrile/water (90:10) by using microwave assisted extraction. The extract is cleaned-up through solid phase extraction with Oasis HLB cartridges and analyzed by ultra high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). Separation is obtained in 3 min. Extraction of analytes from the soil, that is the most critical point, has been studied in detail, and the effect of soil composition and aging time on the analytes recovery has been investigated. PMID- 21641417 TI - Recovery of phosphonate surface contaminants from glass using a simple vacuum extractor with a solid-phase microextraction fiber. AB - Recovery of chemical contaminants from fixed surfaces for analysis can be challenging, particularly if it is not possible to acquire a solid sample to be taken to the laboratory. A simple device is described that collects semi-volatile organic compounds from fixed surfaces by creating an enclosed volume over the surface, then generating a modest vacuum. A solid-phase microextraction (SPME) fiber is then inserted into the evacuated volume where it functions to sorb volatilized organic contaminants. The device is based on a syringe modified with a seal that is used to create the vacuum, with a perforable plunger through which the SPME fiber is inserted. The reduced pressure speeds partitioning of the semi volatile compounds into the gas phase and reduces the boundary layer around the SPME fiber, which enables a fraction of the volatilized organics to partition into the SPME fiber. After sample collection, the SPME fiber is analyzed using conventional gas chromatography/mass spectrometry. The methodology has been used to collect organophosphorus compounds from glass surfaces, to provide a simple test for the functionality of the devices. Thirty minute sampling times (DeltaT(vac)) resulted in fractional recovery efficiencies that ranged from 10( 3) to >10(-2), and in absolute terms, collection of low nanograms was demonstrated. Fractional recovery values were positively correlated to the vapor pressure of the compounds being sampled. Fractional recovery also increased with increasing DeltaT(vac) and displayed a roughly logarithmic profile, indicating that an operational equilibrium is being approached. Fractional recovery decreased with increasing time between exposure and sampling; however, recordable quantities of the phosphonates could be collected three weeks after exposure. PMID- 21641418 TI - A broad-spectrum equine urine screening method for free and enzyme-hydrolysed conjugated drugs with ultra performance liquid chromatography/tandem mass spectrometry. AB - The authors' laboratory at one time employed four liquid chromatography/mass spectrometric (LC/MS) methods for the detection of a large variety of drugs in equine urine. Drug classes covered by these methods included anti-diabetics, anti ulcers, cyclooxygenase-2 (COX-2) inhibitors, sedatives, corticosteroids, anabolic steroids, sulfur diuretics, xanthines, etc. With the objective to reduce labour and instrumental workload, a new ultra performance liquid chromatography/tandem mass spectrometric (UPLC/MS/MS) method has been developed, which encompasses all target analytes detected by the original four LC/MS methods. The new method has better detection limits than the superseded methods. In addition, it covers new target analytes that could not be adequately detected by the four LC/MS methods. The new method involves solid-phase extraction (SPE) of two aliquots of equine urine using two Abs Elut Nexus cartridges. One aliquot of the urine sample is treated with beta-glucuronidase before subjecting to SPE. A second aliquot of the same urine sample is processed directly using another SPE cartridge, so that drugs that are prone to decomposition during enzyme hydrolysis can be preserved. The combined eluate is analysed by UPLC/MS/MS using alternating positive and negative electrospray ionisation in the selected-reaction-monitoring mode. Exceptional chromatographic separation is achieved using an UPLC system equipped with a UPLC((r)) BEH C18 column (10 cm L*2.1 mm ID with 1.7 MUm particles). With this newly developed UPLC/MS/MS method, the simultaneous detection of 140 drugs at ppb to sub-ppb levels in equine urine can be achieved in less than 13 min inclusive of post-run equilibration. Matrix interference for the selected transitions at the expected retention times is minimised by the excellent UPLC chromatographic separation. The method has been validated for recovery and precision, and is being used regularly in the authors' laboratory as an important component of the array of screening methods for doping control analyses of equine urine samples. PMID- 21641419 TI - Highly sensitive detection of clenbuterol using competitive surface-enhanced Raman scattering immunoassay. AB - In this report, we present a novel approach to detect clenbuterol based on competitive surface-enhanced Raman scattering (SERS) immunoassay. Herein, a SERS nanoprobe that relies on gold nanoparticle (GNP) is labeled by 4,4'-dipyridyl (DP) and clenbuterol antibody, respectively. The detection of clenbuterol is carried out by competitive binding between free clenbuterol and clenbuterol-BSA fastened on the substrate with their antibody labeled on SERS nanoprobes. The present method allows us to detect clenbuterol over a much wider concentration range (0.1-100 pg mL(-1)) with a lower limit of detection (ca. 0.1 pg mL(-1)) than the conventional methods. Furthermore, by the use of this new competitive SERS immunoassay, the clenbuterol-BSA (antigen) is chosen to fasten on the substrate instead of the clenbuterol antibody, which could reduce the cost of the assay. Results demonstrate that the proposed method has the wide potential applications in food safety and agonist control. PMID- 21641420 TI - Monitoring of alcoholic fermentation using near infrared and mid infrared spectroscopies combined with electronic nose and electronic tongue. AB - Effective fermentation monitoring is a growing need due to the rapid pace of change in the wine industry, which calls for fast methods providing real time information in order to assure the quality of the final product. The objective of this work is to investigate the potential of non-destructive techniques associated with chemometric data analysis, to monitor time-related changes that occur during red wine fermentation. Eight micro-fermentation trials conducted in the Valtellina region (Northern Italy) during the 2009 vintage, were monitored by a FT-NIR and a FT-IR spectrometer and by an electronic nose and tongue. The spectroscopic technique was used to investigate molecular changes, while electronic nose and electronic tongue evaluated the evolution of the aroma and taste profile during the must-wine fermentation. Must-wine samples were also analysed by traditional chemical methods in order to determine sugars (glucose and fructose) consumption and alcohol (ethanol and glycerol) production. Principal Component Analysis was applied to spectral, electronic nose and electronic tongue data, as an exploratory tool, to uncover molecular, aroma and taste modifications during the fermentation process. Furthermore, the chemical data and the PC1 scores from spectral, electronic nose and electronic tongue data were modelled as a function of time to identify critical points during fermentation. The results showed that NIR and MIR spectroscopies are useful to investigate molecular changes involved in wine fermentation while electronic nose and electronic tongue can be applied to detect the evolution of taste and aroma profile. Moreover, as demonstrated through the modeling of NIR, MIR, electronic nose and electronic tongue data, these non destructive methods are suitable for the monitoring of must-wine fermentation giving crucial information about the quality of the final product in agreement with chemical parameters. Although in this study the measurements were carried out in off-line mode, in future these non destructive techniques could be valid and simple tools, able to provide in time information about the fermentation process and to assure the quality of wine. PMID- 21641421 TI - Integration of fiber optic-particle plasmon resonance biosensor with microfluidic chip. AB - This article reports the integration of the fiber optic-particle plasmon resonance (FO-PPR) biosensor with a microfluidic chip to reduce response time and improve detection limit. The microfluidic chip made of poly(methyl methacrylate) had a flow-channel of dimensions 4.0 cm * 900 MUm * 900 MUm. A partially unclad optical fiber with gold or silver nanoparticles on the core surface was placed within the flow-channel, where the volume of the flow space was about 14 MUL. Results using sucrose solutions of various refractive indexes show that the refractive index resolution improves by 2.4-fold in the microfluidic system. The microfluidic chip is capable of delivering a precise amount of biological samples to the detection area without sample dilution. Several receptor/analyte pairs were chosen to examine the biosensing capability of the integrated platform: biotin/streptavidin, biotin/anti-biotin, DNP/anti-DNP, OVA/anti-OVA, and anti-MMP 3/MMP-3. Results show that the response time to achieve equilibrium can be shortened from several thousand seconds in a conventional liquid cell to several hundred seconds in a microfluidic flow-cell. In addition, the detection limit also improves by about one order of magnitude. Furthermore, the normalization by using the relative change of transmission response as the sensor output alleviate the demand on precise optical alignment, resulting in reasonably good chip-to chip measurement reproducibility. PMID- 21641422 TI - Curcumin induced nanoscale CD44 molecular redistribution and antigen-antibody interaction on HepG2 cell surface. AB - The cell surface glycoprotein CD44 was implicated in the progression, metastasis and apoptosis of certain human tumors. In this study, we used atomic force microscope (AFM) to monitor the effect of curcumin on human hepatocellular carcinoma (HepG2) cell surface nanoscale structure. High-resolution imaging revealed that cell morphology and ultrastructure changed a lot after being treated with curcumin. The membrane average roughness increased (10.88 +/- 4.62 nm to 129.70 +/- 43.72 nm) and the expression of CD44 decreased (99.79 +/- 0.16% to 75.14 +/- 8.37%). Laser scanning confocal microscope (LSCM) imaging showed that CD44 molecules were located on the cell membrane. The florescence intensity in control group was weaker than that in curcumin treated cells. Most of the binding forces between CD44 antibodies and untreated HepG2 cell membrane were around 120-220 pN. After being incubated with curcumin, the major forces focused on 70-150 pN (10 MUM curcumin-treated) and 50-120 pN (20 MUM curcumin-treated). These results suggested that, as result of nanoscale molecular redistribution, changes of the cell surface were in response to external treatment of curcumin. The combination of AFM and LSCM could be a powerful method to detect the distribution of cell surface molecules and interactions between molecules and their ligands. PMID- 21641423 TI - Enhanced electrochemiluminescence from luminol at multi-walled carbon nanotubes decorated with palladium nanoparticles: a novel route for the fabrication of an oxygen sensor and a glucose biosensor. AB - Incorporation of palladium nanoparticles on the surface of multi-walled carbon nanotubes and modification of glassy carbon electrode with the prepared nano hybrid material led to the fabrication of a novel electrode. The modified electrode showed attractive electrocatalytic activity and sensitizing effect on luminol-O(2) and luminol-H(2)O(2) electrochemiluminescence (ECL) reactions at neutral media. The sensitized luminol-O(2) and luminol-H(2)O(2) reactions were successfully applied for the ECL determination of dissolved O(2) and glucose, respectively. Under the optimal conditions for luminol-O(2) system, the ECL signal intensity of luminol was linear with the concentration of dissolved oxygen in the range between 0.08 and 0.94 mM (r=0.9996) and for luminol-H(2)O(2) system, the ECL signal intensity of luminol was linear with the concentration of glucose in the range between 0.1 and 1000 MUM (r=0.9998). The limits of detection (S/N=3) for dissolved oxygen and glucose were 0.02 mM and 54 nM, respectively. The relative standard deviations (RSD) for repetitive measurements of 0.50 mM oxygen (n=10) and 10 MUM glucose (n=30) were 3.5% and 0.3%, respectively. Also, under the optimal conditions for luminol-H(2)O(2) system, the ECL signal intensity of luminol was linear with the concentration of H(2)O(2) in the range between 1 nM and 0.45 mM (r=0.9997). The limit of detection (S/N=3) for H(2)O(2) detection was 0.5 nM and the relative standard deviation for repetitive measurements of 10 MUM H(2)O(2) (n=10) was 0.8%. PMID- 21641425 TI - Sixteenth International Conference on Flow Injection Analysis Including Related Techniques (ICFIA 2010). PMID- 21641424 TI - Chronocoulometric determination of urea in human serum using an inkjet printed biosensor. AB - A biosensor for the determination of urea in human serum was fabricated using a combination of inkjet printed polyaniline nanoparticles and inkjet printed urease enzyme deposited sequentially onto screen-printed carbon paste electrodes. Chronocoulometry was used to measure the decomposition of urea via the doping of ammonium at the polyaniline-modified electrode surface at -0.3 V vs. Ag/AgCl. Ammonium could be measured in the range from 0.1 to 100 mM. Urea could be measured by the sensor in the range of 2-12 mM (r(2)=0.98). The enzyme biosensor was correlated against a spectrophotometric assay for urea in 15 normal human serum samples which yielded a correlation coefficient of 0.85. Bland-Altman plots showed that in the range of 5.8-6.6 mM urea, the developed sensor had an average positive experimental bias of 0.12 mM (<2% RSD) over the reference method. PMID- 21641426 TI - How did flow injection analysis, and its related techniques, develop in various parts of the globe? Reflections of prominent FIA practitioners. PMID- 21641427 TI - Flow injection spectrophotometric determination of formaldehyde based on its condensation with hydroxylamine and subsequent redox reaction with iron(III) ferrozine complex. AB - A flow injection (FI) spectrophotometric method is proposed for the determination of low concentration of formaldehyde (HCHO) in liquid media. It is based on the condensation of HCHO with hydroxylamine sulfate, followed by the reduction reaction of iron(III)-ferrozine complex with the residual hydroxylamine to form a purple iron(II)-ferrozine complex (lambda(max)=562 nm). In the first reaction, hydroxylamine decreases proportionally to the concentration of HCHO, and therefore the produced purple iron(II)-ferrozine complex decreases with increasing HCHO (a negative FI peak is obtained). The detection limit (S/N=3) was 1.6 MUg L(-1). The method can be applied to the determination of HCHO in industrial wastewater. PMID- 21641428 TI - Determination of uranium in seawater by flow-injection preconcentration on dodecylamidoxime-impregnated resin and spectrophotometric detection. AB - A flow injection method has been developed for the determination of uranium in seawater combining the on-line preconcentration with spectrophotometric detection. An aliquot (10 mL) of the seawater sample adjusted to pH 5.5 was injected into the analytical system and uranium was adsorbed on the column packed with styrene-divinylbenzene copolymer resin (Bio-Beads SM-2) modified with dodecylamidoxime which showed high selectivity to uranium. Uranium was then eluted with 0.01 M hydrochloric acid and detected spectrophotometrically after the reaction with Chlorophosphonazo III. Interference from calcium and strontium was masked with cyclohexanediaminetetraacetic acid added to the chromogenic reagent solution. The sample throughput, the detection limit (3sigma), and the preconcentration factor were 23 per hour, 0.13 MUg/L, and 20, respectively, when the sample injection volume was kept at 10 mL. The precision at the 2 MUg/L level was less than 4% (RSD). The proposed method was applied to the determination of uranium in the seawater samples collected off the Boso peninsula, Japan and the uranium concentration was found to be ca. 3 MUg/L, which is close to the literature data. The yield of the recovery test ranged from 95% to 99%. PMID- 21641429 TI - Sequential injection dispersive liquid-liquid microextraction based on fatty alcohols and poly(etheretherketone)-turnings for metal determination by flame atomic absorption spectrometry. AB - A novel, simple and efficient sequential injection (SI) on-line dispersive liquid liquid microextraction (DLLME) procedure was described and was demonstrated for the assay of trace silver determination by flame atomic absorption spectrometry (FAAS). Fatty alcohols, such as 1-undecanol and 1-dodecanol, were examined as extraction solvents at microlitre volume, overcoming a major problem of the DLLME methods, the high toxicity of the extraction solvents used. Furthermore, the extractant fine droplets can be easily separated from the aqueous phase using a micro-column packed with a novel hydrophobic sorbent material, poly(etheretherketone)-turnings. In this method fine droplets of 1-dodecanol were on-line generated and dispersed into the stream of aqueous sample. By this continuous process, silver diethyldithiocarbamate (Ag-DDTC) complex was formed and extracted into the dispersed extraction solvent. No specific conditions such as ice bath for low temperature or special tools are required for extractant isolation. All significant parameters that influence the efficiency of the system such as sample acidity, concentration of complexing reagent and extraction solvent, flow-rate of disperser and sample solution as well as the preconcentration time were investigated and optimized by full factorial design. Under the optimized conditions a detection limit of 0.15 MUg L(-1), a relative standard deviation (RSD) of 2.9% at 5.00 MUg L(-1) Ag(I) concentration level and an enhancement factor of 186 were obtained. The developed method was evaluated by analyzing certified reference material and was applied successfully to the analysis of environmental water samples. PMID- 21641430 TI - Lab on valve-multisyringe flow injection system (LOV-MSFIA) for fully automated uranium determination in environmental samples. AB - The hyphenation of lab-on-valve (LOV) and multisyringe flow analysis (MSFIA), coupled to a long path length liquid waveguide capillary cell (LWCC), allows the spectrophotometric determination of uranium in different types of environmental sample matrices, without any manual pre-treatment, and achieving high selectivity and sensitivity levels. On-line separation and preconcentration of uranium is carried out by means of UTEVA resin. The potential of the LOV-MSFIA makes possible the fully automation of the system by the in-line regeneration of the column. After elution, uranium(VI) is spectrophotometrically detected after reaction with arsenazo-III. The determination of levels of uranium present in environmental samples is required in order to establish an environmental control. Thus, we propose a rapid, cheap and fully automated method to determine uranium(VI) in environmental samples. The limit of detection reached is 1.9 etag of uranium and depending on the preconcentrated volume; it results in ppt levels (10.3 etag L(-1)). Different water sample matrices (seawater, well water, freshwater, tap water and mineral water) and a phosphogypsum sample (with natural uranium content) were satisfactorily analyzed. PMID- 21641431 TI - Pressure-assisted capillary electrophoresis for cation separations using a sequential injection analysis manifold and contactless conductivity detection. AB - Pressure assisted capillary electrophoresis in capillaries with internal diameters of 10 MUm was found possible without significant penalty in terms of separation efficiency and sensitivity when using contactless conductivity detection. A sequential injection analysis manifold consisting of a syringe pump and valves was used to impose a hydrodynamic flow in the separation of some inorganic as well as organic cations. It is demonstrated that the approach may be used to optimize analysis time by superimposing a hydrodynamic flow parallel to the electrokinetic motion. It is also possible to improve the separation by using the forced flow to maintain the analytes in the capillary, and thus the separation field, for longer times. The use of the syringe pump allows flexible and precise control of the pressure, so that it is possible to impose pressure steps during the separation. The use of this was demonstrated for the speeding up of late peaks, or forcing repeated passage of the sample plug through the capillary in order to increase separation. PMID- 21641432 TI - Examination of colour inkjet printing inks by capillary electrophoresis. AB - The possibility of comparing inkjet printing inks by micellar electrokinetic capillary electrophoresis (MECC) with diode array detection was studied. An analytical procedure was designed and successfully applied to discriminate between the electrophoretic profiles of inks (extracted from paper) produced by five well-known manufacturers. The separation process was conducted in a polyimide-coated fused silica capillary (ID 50 MUm, 60 cm total/50 cm effective length) with +30 kV high voltage applied. Background electrolyte was used of the following optimum composition: 40 mM sodium borate buffer, 20mM sodium dodecyl sulphate(IV) (SDS) and 10% (v/v) acetonitrile (pH 9.56). The experimental conditions were adjusted in terms of resolution and analysis time. The best results were obtained at 10 and 25 degrees C storage and capillary temperature, respectively, using 25 dots (o 0.8mm) cut from printouts as the sample and BGE diluted with water (1:99, v/v) as the injecting solution. The MECC separation of main printing ink components by the proposed method showed excellent precision - the RSD value of the migration time calculated for each of the investigated peaks did not exceed 3.3%. The optimized method was applied to group identification and differentiation of: (a) three colours of printing inks, (b) inks from different manufacturers (Hewlett-Packard, Epson, Brother, Lexmark and Canon) and (c) inks from different printer models. In all these cases, inks were successfully differentiated on the basis of position (migration time) and shape of their characteristic peaks. PMID- 21641433 TI - A membraneless gas-diffusion unit-multisyringe flow injection spectrophotometric method for ammonium determination in untreated environmental samples. AB - A new design of a membraneless gas-diffusion (MGD) unit coupled to a multisyringe flow injection system is proposed. The spectrophotometric determination of ammonium using an acid-base indicator was chosen to show the feasibility of this approach. Hence, in alkaline medium, ammonium ions are transformed into ammonia (donor channel) which diffuses through the headspace into the acceptor stream (bromothymol blue solution), causing a pH change and subsequently a colour change. The exploitation of the enhanced potentialities of this re-designed MGD device was the main purpose of the present work. Hence, several strategies concerning flow management were studied seeking to characterize and improve the analytical features of the methodology and moreover, untreated environmental samples were analysed without previous filtration. Consequently, stopped flow in acceptor channel with continuous flow in donor channel was chosen for the application to wastewater and spiked river water samples. A linear concentration range between 10.0 and 50.0 mg L(-1) of NH(4)(+), a limit of detection of 2.20 mg L(-1) and a determination frequency of 11h(-1) were obtained. PMID- 21641434 TI - Room temperature imidazolium ionic liquid: a solvent for extraction of carbamates prior to liquid chromatographic analysis. AB - A simple and rapid method for preconcentration of carbamate insecticides, including methomyl, propoxur, carbofuran, carbaryl, isoprocarb, methiocarb and promecarb, has been developed. It was based on a liquid-liquid microextraction using a [C(4)MIM][PF(6)] room temperature ionic liquid as an extraction solvent prior to analysis by high performance liquid chromatography with UV detection. Experimental parameters affecting the extraction performance, such as the volumes of sample, extractant and dissolving solvent, and extraction time, were studied. Under the selected conditions, the enrichment factors in the range between 10 and 25 could be achieved with the limit of detection in the range of 2-40 MUg L(-1), and with the relative standard deviations of lower than 0.6 and 10.2% for retention time and peak area, respectively. The proposed method offers advantages in reduction of the exposure danger to toxic solvents used in the conventional liquid-liquid extraction, simplicity of the extraction processes, rapidity, and sensitivity enhancement. The method was demonstrated to apply to the analysis of fruit and natural surface water samples. PMID- 21641435 TI - Automated flow-based anion-exchange method for high-throughput isolation and real time monitoring of RuBisCO in plant extracts. AB - In this work, a miniaturized, completely enclosed multisyringe-flow system is proposed for high-throughput purification of RuBisCO from Triticum aestivum extracts. The automated method capitalizes on the uptake of the target protein at 4 degrees C onto Q-Sepharose Fast Flow strong anion-exchanger packed in a cylindrical microcolumn (105 * 4 mm) followed by a stepwise ionic-strength gradient elution (0-0.8 mol/L NaCl) to eliminate concomitant extract components and retrieve highly purified RuBisCO. The manifold is furnished downstream with a flow-through diode-array UV/vis spectrophotometer for real-time monitoring of the column effluent at the protein-specific wavelength of 280 nm to detect the elution of RuBisCO. Quantitation of RuBisCO and total soluble proteins in the eluate fractions were undertaken using polyacrylamide gel electrophoresis (PAGE) and the spectrophotometric Bradford assay, respectively. A comprehensive investigation of the effect of distinct concentration gradients on the isolation of RuBisCO and experimental conditions (namely, type of resin, column dimensions and mobile-phase flow rate) upon column capacity and analyte breakthrough was effected. The assembled set-up was aimed to critically ascertain the efficiency of preliminary batchwise pre-treatments of crude plant extracts (viz., polyethylenglycol (PEG) precipitation, ammonium sulphate precipitation and sucrose gradient centrifugation) in terms of RuBisCO purification and absolute recovery prior to automated anion-exchange column separation. Under the optimum physical and chemical conditions, the flow-through column system is able to admit crude plant extracts and gives rise to RuBisCO purification yields better than 75%, which might be increased up to 96 +/- 9% with a prior PEG fractionation followed by sucrose gradient step. PMID- 21641436 TI - Spectrophotometric determination of zinc and copper in a multi-syringe flow injection analysis system using a liquid waveguide capillary cell: application to natural waters. AB - This work exploits a multi-syringe injection analysis (MSFIA) system coupled with a long liquid waveguide capillary cell for the spectrophotometric determination of zinc and copper in waters. A liquid waveguide capillary cell (1.0m pathlength, 550 MUm i.d. and 250 MUL internal volume) was used to enhance the sensitivity of the detection. The determination for both ions is based on a colorimetric reaction with zincon at different pH values. The developed methodology compares favourably with other previously described procedures, as it allows to reach low detection limits for both cations (LODs of 0.1 and 2 MUg L(-1), for copper and zinc, respectively), without the need for any pre-concentration step. The system also provided a linear response up to 100 MUg L(-1) with a high throughput (43 h( 1)) and low reagent consumption and effluent production. The developed work was applied to natural waters and three certified reference water samples. PMID- 21641437 TI - Simple automated generation of gradient elution conditions in sequential injection chromatography using monolithic column. AB - The paper deals with the concept of simple automated creation of gradient profile of the mobile phase for gradient-elution sequential injection chromatography (GE SIC). The feasibility and merits of this concept are demonstrated on the separation and simultaneous assay of indomethacin as active principle and of its two degradation products (5-methoxy-2-methylindoleacetic acid and 4-chloro benzoic acid) in a topical pharmaceutical formulation. The GE-SIC separation was performed with a FIAlab((r)) 3000 SIC set-up (USA) equipped with an OnyxTM Monolithic C18 (25 mm * 4.6mm, Phenomenex((r))) column, a six-port selection valve, a 5-mL syringe pump and a fiber-optics UV CCD detector. Ketoprofen was used as an internal standard (IS). The gradient elution was achieved by automated reproducible mixing of acetonitrile and aqueous 0.2% phosphoric acid in the holding coil of the SIC system. Different profiles of the gradient elution were tested. The optimal gradient using two mobile phases 30:70 and 50:50 of acetonitrile/0.2% phosphoric acid (v/v) was achieved under the optimum flow rate 1.2 mL min(-1). The chromatographic resolution R between the peaks of all solutes (including the IS) was >2.00. The repeatability of retention times was characterized by the RSD values 0.18-0.30% (n=6). Net separation time was 3.5 min and the mobile phase consumption was 4.5 mL for a single GE-SIC assay. The figures of merit of the novel GE-SIC method compared well with those of conventional HPLC. PMID- 21641438 TI - The use of a polymer inclusion membrane in flow injection analysis for the on line separation and determination of zinc. AB - This paper reports the first use of a polymer inclusion membrane (PIM) for on line separation in flow injection analysis (FIA) involving simultaneous extraction and back-extraction. The FIA system containing the PIM separation module was used for the determination of Zn(II) in aqueous samples in the presence of Mg(II), Ca(II), Cd(II), Co(II), Ni(II), Cu(II), and Fe(III). The Fe(III) and Cu(II) interferences were eliminated by off-line precipitation with phosphate and on-line complexation with chloride, respectively. The concentration of Zn(II) was determined spectrophotometrically using 4-(2-pyridylazo) resorcinol (PAR). The optimal composition of the PIM consisted of 40% (m/m) di(2-ethlyhexyl) phosphoric acid (D2EHPA) as carrier, 10% (m/m) dioctyl phthalate (DOP) as plasticizer and 50% (m/m) poly(vinyl chloride) (PVC) as the base polymer. The optimized FIA system was characterized by a linear calibration curve in the range from 1.0 to 30.0 mg L(-1) Zn(II), a detection limit of 0.05 mg L(-1) and a relative standard deviation of 3.4% with a sampling rate of 4h(-1). Reproducible results were obtained for 20 replicate injections over a 5h period which demonstrated a good membrane stability. The FIA system was applied to the determination of Zn(II) in pharmaceuticals and samples from the galvanizing industry and very good agreement with atomic absorption spectrometry was obtained. PMID- 21641439 TI - Surface plasmon resonance for real-time study of lectin-carbohydrate interactions for the differentiation and identification of glycoproteins. AB - A study of specific interactions between lectins and glycoproteins has been carried out using surface plasmon resonance (SPR) in a flow-injection mode. Lectins were covalently immobilised on the surfaces of the microfluidic sensor chip via amine coupling and serum glycoproteins were injected into the flow channels. Specific lectin-glycoprotein interactions caused the shift of refractive index proportional to the mass concentration accumulated on the channel surface. Lectins showed different affinity to the tested glycoproteins and each glycoprotein displayed its own lectin-binding pattern. It is possible to distinguish and identify even glycoproteins with similar sugar structures by simple and quick screening. The working conditions of the assay were optimised. The lectin-based SPR made it possible to carry out the label-free detection of glycoproteins within a broad concentration range with a good linearity. Regeneration conditions for the surface of the sensor chip were found and optimised. Combination of 10mM HCl and 10mM glycine-HCl (pH 2.5) removes the bound glycoproteins from the lectin surface without damaging it. The kinetic and affinity parameters of lectin-glycoprotein binding were evaluated. The proposed method was tested on human glycosylated serum. Combination of the lectin panel with SPR is suitable both for specific screening and for sensitive assay of serum glycoproteins. PMID- 21641440 TI - Development of flow injection potentiometric methods for the off-line and on-line determination of fluoride to monitor the biodegradation of a monofluorophenol in two bioreactors. AB - Water treatment has become a source of concern as new pollutants and higher volumes of waste water must be treated. Emerging biological approaches, namely the use of bioreactors, for cleaning processes have been introduced. The use of bioreactors requires the development of efficient monitoring tools, preferably with real-time measurements. In this work, a couple of flow injection systems were developed and optimized for the potentiometric determination of fluoride to monitor a rotating biological contactor (RBC) bioreactor and a sequencing batch reactor (SBR) with off-line and on-line sampling. Both the RBC and the SBR bioreactors were set up for the biodegradation of the halogenated organic compound 2-fluorophenol and, as fluoride was a degradation byproduct, the process was monitored by following up its concentration. The described flow injection potentiometric methods enabled the fluoride determination within the required quantification range 0.10-100mM. The possible interferences from the growth medium were minimized in-line. The determination rate was 78 h(-1) for the off line monitoring of RBC and 50(-1)h for the on-line monitoring of the SBR, with a sample consumption of 0.500 mL and 0.133 mL per determination, respectively. Furthermore, the overall reagent consumption was quite low. The accuracy of the system was evaluated by comparison with a batch procedure. The SBR efficiency was monitored both on-line by the flow system and off-line by HPLC, for comparison purposes. PMID- 21641441 TI - Exploiting the bead injection LOV approach to carry out spectrophotometric assays in wine: application to the determination of iron. AB - A sequential injection lab-on-valve (SI-LOV) system was used to develop a new methodology for the determination of iron in wine samples exploiting the bead injection (BI) concept for solid phase extraction and spectrophotometric measurement. Nitrilotriacetic Acid (NTA) Superflow resin was used to build the bead column of the flow through sensor. The iron (III) ions were retained by the bead column and react with SCN(-) producing an intense red colour. The change in absorbance was monitored spectrophotometrically on the optosensor at 480 nm. It was possible to achieve a linear range of 0.09-5.0 mg L(-1) of iron, with low sample and reagent consumption; 500 MUL of sample, 15 MUmol of SCN(-), and 9 MUmol of H(2)O(2), per assay. The proposed method was successfully applied to the determination of iron in wine, with no previous treatment other than dilution, and to other food samples. PMID- 21641442 TI - Single strand hollow fiber membrane (SSHFM): an on-line sample preparation for the flow based colorimetric determination of free iron in fruit juices. AB - A single strand hollow fiber membrane (SSHFM) was developed for the on-line sample preparation for the flow based colorimetric determination of free iron levels in fruit juices. The SSHFM, as used, could separate Fe(2+) from some spectrophotometric interfering agents in the fruit juice, such as pigments, solid suspensions and polysaccharides. The screening process was likely to have been primarily based on dialysis, wherein only ions or molecules that are smaller than the pores of the membrane can diffuse through while relatively larger molecules or particles could not. Two flow modes, a continuous and stopped flow, were studied. Factors that influenced the sensitivity (%dialysis) of the method, such as the flow rate, sample volume, flow direction and stopped flow time, were optimized. The stopped flow mode was found to be relatively more sensitive than the continuous flow mode and displayed a linear range of 1-30 mg L(-1) Fe(2+), a limit of detection of 0.5 mg L(-1), and a % relative standard deviation of less than 2% (n=8) for the analysis of 10 mg L(-1) Fe(2+) spiked grape juice samples. A sample throughput of 24 samples h(-1), was attained without any further sample treatment. PMID- 21641443 TI - A reagent-free method based on a photo-induced fluorimetry in a sequential injection system. AB - According to the current demands of environmentally friendly analytical chemistry and with a view to achieving lower reagent consumption with improved analytical performance, an automatic methodology composed of a photoreactor and fluorimetric detection (lambda(exc)=287 nm, lambda(em)=378 nm) was developed. To this end, a sequential injection analysis (SIA) system was developed for indomethacin determination using ultra-violet (UV) light which promotes an increase in the fluorescence of indomethacin. This increase in sensitivity makes it possible to apply this methodology to a dissolution test and to determine indomethacin in pharmaceutical formulations. The calibration graph for indomethacin was linear between 4.10 * 10(-6) and 9.00 * 10(-5) mol L(-1)and the detection limit was 1.23 * 10(-6) mol L(-1). The method was proven to be reproducible with a R.S.D.<5% and sampling rate of approximately 20 per hour. The potential effect of several compounds commonly used as excipients on analytical signals was studied and no interfering effect was observed. Statistical evaluation at the 95% confidence level showed good agreement between the results obtained for the pharmaceutical samples with both the SIA system and comparison batch procedures. PMID- 21641444 TI - Cadmium telluride nanocrystals as luminescent sensitizers in flow analysis. AB - A fully automated multipumping flow system (MPFS) using water-soluble CdTe quantum dots (QD) as sensitizers is proposed for the chemiluminometric determination of the anti-diabetic drugs gliclazide and glipizide in pharmaceutical formulations. The nanocrystals acted as enhancers of the weak CL emission produced upon oxidation of sulphite by Ce(IV) in acidic medium, thus improving sensitivity and expanding the dynamical analytical concentration range. By interacting with the QD, the two analytes prevented their sensitizing effect yielding a chemiluminescence quenching of the Ce(IV)-SO(3)(2-)CdTe QD system. The pulsed flow inherent to MPFS assured a fast and efficient mixing of all solutions inside the flow cell, circumventing the need for a reaction coil and facilitating the monitoring of the short-lived generated chemiluminescent species. QD crystal size, concentration and spectral region for measurement were investigated. PMID- 21641445 TI - Flow injection analysis combined with a hydrothermal flow reactor: application to kinetic determination of trace amounts of iridium using a water-soluble porphyrin. AB - A new type of flow injection analysis (FIA) system combined with an extremely high temperature reactor, namely hydrothermal flow injection analysis (HT-FIA), has been successfully constructed for the first time. Fundamental characteristics of HT-FIA system, such as limit temperature, pressure, and flow rate, were examined as an analytical tool. To demonstrate the potential of HT-FIA, the catalytic activity of Ir(IV) for the degradation of a water-soluble porphyrin, 5,10,15,20-tetraphenyl-21H,23H-porphinetetrasulfonic acid (TPPS), was applied for the determination of trace amounts of Ir(IV). Although the indicator reaction is very slow at room temperature, HT-FIA system enables to accelerate the reaction. A linear calibration curve was acquired at 10(-8)M level of Ir(IV) and the interferences of platinum group metal ions were examined. The detection limit of Ir(IV) was 5.8 * 10(-9)M and a fairly high-throughput analysis, of which more than 30 samples can be analyzed within 80 min, was achieved. PMID- 21641446 TI - A fast and highly sensitive detection of cholesterol using polymer microfluidic devices and amperometric system. AB - In this work, the rapid detection of cholesterol using poly(dimethylsiloxane) microchip capillary electrophoresis, based on the coupling of enzymatic assays and electrochemical detection, was developed. Direct amperometric detection for poly(dimethylsiloxane) (PDMS) microchip capillary electrophoresis was successfully applied to quantify cholesterol levels. Factors influencing the performance of the method (such as the concentration and pH value of buffer electrolyte, concentration of cholesterol oxidase enzyme (ChOx), effect of solvent on the cholesterol solubility, and interferences) were carefully investigated and optimized. The migration time of hydrogen peroxide, product of the reaction, was less than 100 s when using 40 mM phosphate buffer at pH 7.0 as the running buffer, a concentration of 0.68 U/mL of the ChOx, a separation voltage of +1.6 kV, an injection time of 20s, and a detection potential of +0.5 V. PDMS microchip capillary electrophoresis showed linearity between 38.7 MUg/dL (1 MUM) and 270.6 mg/dL (7 mM) for the cholesterol standard; the detection limit was determined as 38.7 ng/dL (1 nM). To demonstrate the potential of this assay, the proposed method was applied to quantify cholesterol in bovine serum. The percentages of recoveries were assessed over the range of 98.9-101.8%. The sample throughput was found to be 60 samples per hour. Therefore, PDMS microchip capillary electrophoresis, based on the coupling of enzymatic assays and electrochemical detection, is very rapid, accurate and sensitive method for the determination of cholesterol levels. PMID- 21641447 TI - Automatic miniaturized fluorometric flow system for chemical and toxicological control of glibenclamide. AB - In this work, and for the first time, it was developed an automatic and fast screening miniaturized flow system for the toxicological control of glibenclamide in beverages, with application in forensic laboratory investigations, and also, for the chemical control of commercially available pharmaceutical formulations. The automatic system exploited the multipumping flow (MPFS) concept and allowed the implementation of a new glibenclamide determination method based on the fluorometric monitoring of the drug in acidic medium (lambda(ex)=301 nm; lambda(em)=404 nm), in the presence of an anionic surfactant (SDS), promoting an organized micellar medium to enhance the fluorometric measurements. The developed approach assured good recoveries in the analysis of five spiked alcoholic beverages. Additionally, a good agreement was verified when comparing the results obtained in the determination of glibenclamide in five commercial pharmaceutical formulations by the proposed method and by the pharmacopoeia reference procedure. PMID- 21641448 TI - Measurements of arsenite and arsenate contained in mining river waters and leached from contaminated sediments by sequential hydride generation flow injection analysis. AB - In this work, a new analytical method for gasifiable compounds based on sequential hydride generation flow injection analysis (SHGFIA) was applied to water analysis and leaching investigation. For water analysis, it was confirmed that 1 MUg L(-1) As(III) and As(V) were stable for a few days when EDTA was added in the sample waters. Dissolved As(III) and total arsenic (As(III)+As(V)) were converted to AsH(3) in neutral and acidic medium, respectively, to transfer to a miniature gas scrubber (100 MUL in absorber volume). The collected arsenic was successively measured by flow analysis based on molybdenum blue chemistry. With this system, changes in As(III) and As(V) concentrations of water placed with arsenic-contaminated-sediment was monitored in near real time. From these data, kinetic analyses were carried out and kinetic constant was obtained from plot of ln{(C(infinity)-C)/C(infinity)} where C and C(infinity) were leached arsenic concentration and its final concentration, respectively. It was found that rate of As(III) leaching was much faster than that of As(V) while As(V) leached more in amount compared to As(III). In this work, it was demonstrated that kinetic investigation is also one of the important application of flow analysis. The SHGFIA system showed excellent performance for leaching analysis of arsenic with discrimination of As(III) and As(V). PMID- 21641449 TI - Simultaneous determination of some food additives in soft drinks and other liquid foods by flow injection on-line dialysis coupled to high performance liquid chromatography. AB - Flow injection on-line dialysis was developed for sample pretreatment prior to the simultaneous determination of some food additives by high performance liquid chromatography (FID-HPLC). A liquid sample or mixed standard solution (900 MUL) was injected into a donor stream (5%, w/v, sucrose) of FID system and was pushed further through a dialysis cell, while an acceptor solution (0.025 mol L(-1) phosphate buffer, pH 3.75) was held in the opposite side of the dialysis membrane. The dialysate was then flowed to an injection loop of the HPLC valve, where it was further injected into the HPLC system and analyzed under isocratic reverse-phase HPLC conditions and UV detection (230 nm). The order of elution of five food additives was acesulfame-K, saccharin, caffeine, benzoic acid and sorbic acid, respectively, with the analysis time of 14 min. On-line dialysis and HPLC analysis could be performed in parallel, providing sample throughput of 4.3h(-1). Dialysis efficiencies of five food additives were in ranges of 5-11%. Linear calibration graphs were in ranges of 10-100 mg L(-1) for acesulfame-K and saccharin, 10-250 mg L(-1) for benzoic acid and 10-500 mg L(-1) for caffeine and sorbic acid. Good precisions (RSD<5%) for all the additives were obtained. The proposed system was applied to soft drink and other liquid food samples. Acceptable percentage recoveries could be obtained by appropriate dilution of the sample before injecting into the system. The developed system has advantages of high degrees of automation for sample pretreatment, i.e., on-line sample separation and dilution and low consumption of chemicals and materials. PMID- 21641450 TI - Sequential injection analysis with electrochemical detection as a tool for economic and rapid evaluation of total antioxidant capacity. AB - This work presents a new flow-based coupled electrochemical technique for evaluation of "total antioxidant capacity (TAC)". A sequential injection (SI) with amperometric detection was applied to the TAC analysis of commercial instant ginger infusion beverages using 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay. Besides having chromogenic properties, the ABTS reagent behaves as an electroactive species at the glassy carbon electrode in phosphate buffer pH 7.0, the decrease of the cathodic current signal of the ABTS(+) radical after reaction with antioxidants can be monitored. The SI system, furnished with an in-house electrochemical detection cell (ECD), was optimized with respect to the applied potential, sample and reagent volume, and flow rate to the detector. Gallic acid was used as the standard antioxidant and the capacity was reported as gallic acid equivalent (GAE) unit. TAC measurements of ginger infusions at the optimum condition were performed using the proposed technique and also with the classical batch spectrophotometric ABTS assay. TAC values obtained from our method and the standard method are in good agreement (r(2)=0.956). The SI amperometric technique provided satisfactory precision (4.11% RSD) with rapid sample throughput (40 samples h(-1)). Also using this method, the consumption of the expensive ABTS reagent was greatly reduced. PMID- 21641451 TI - Sequential injection determination of orthophosphate as ion associate of 12 molybdophosphate with Astra Phloxine. AB - A simple and fast reaction between 12-molybdophosphate heteropoly anion and the polymethine dye Astra Phloxine was used for the development of a new SIA method for the determination of orthophosphate. The suggested method has higher sensitivity and a broader calibration range than existing SIA methods. It can be used to detect phosphate in the range from 0.2 to 10 MUmol L(-1) with a detection limit of 0.1 MUmol L(-1) and an acceptable throughput of 20 samples h(-1). The comparably low molybdate (5.6 mmol L(-1)) and dye (0.1 mmol L(-1)) concentrations led to an improvement in the stability of the base line. Inter-day reproducibility was better than 5%, while the intra-day RSD was in the range 0.8 3.5%. The method was used for the analysis of natural water samples and phosphorus-containing chemicals having a low content of orthophosphate. PMID- 21641452 TI - Sensitive fluorimetric flow injection analysis for fluoride ion with a novel reagent, 2',7'-dichlorofluorescein di-tert-butyldimethylsilyl ether. AB - A novel reagent, 2',7'-dichlorofluorescein di-tert-butyldimethylsilyl ether (FCl(2)TBS), was synthesized for fluoride ion and used for a sensitive fluorimetric flow injection analysis by detecting the recovery of fluorescence due to cleavage of Si-O bond. Four kinds of fluorescein di-tert butyldimethylsilyl ether (FTBS) analogues were synthesized and FCl(2)TBS was the best. By introducing chlorine to FTBS, stability of the reagent, reactivity and the baseline signals were improved. The FIA system was three lines. The sample solution (aqua medium) was injected in the carrier solution (water) and merged with the reagent solution (2.0 * 10(-5)M FCl(2)TBS acetone solution), then mixed with phosphate buffer solution (pH 7.5). The fluorescence intensities were measured at lambda(ex) 503 nm and lambda(em) 527 nm. The calibration graph had linear relationship between (1.0-50.0) * 10(-6)M and the determinable limit was 1.0 * 10(-6)M. The relative standard deviation of 12 measurements with 1.0 * 10( 5)M F(-) solution was 1.0% and the sample throughput was 13 h(-1). The developed method was successfully applied to river and tap water samples. PMID- 21641453 TI - Sequential injection anodic stripping voltammetry with monosegmented flow and in line UV digestion for determination of Zn(II), Cd(II), Pb(II) and Cu(II) in water samples. AB - A cost-effective sequential injection system incorporating with an in-line UV digestion for breakdown of organic matter prior to voltammetric determination of Zn(II), Cd(II), Pb(II) and Cu(II) by anodic stripping voltammetry (ASV) on a hanging mercury drop electrode (HMDE) of a small scale voltammetric cell was developed. A low-cost small scale voltammetric cell was fabricated from disposable pipet tip and microcentrifuge tube with volume of about 3 mL for conveniently incorporated with the SI system. A home-made UV digestion unit was fabricated employing a small size and low wattage UV lamps and flow reactor made from PTFE tubing coiled around the UV lamp. An in-line single standard calibration or a standard addition procedure was developed employing a monosegmented flow technique. Performance of the proposed system was tested for in-line digestion of model water samples containing metal ions and some organic ligands such as strong organic ligand (EDTA) or intermediate organic ligand (humic acid). The wet acid digestion method (USEPA 3010a) was used as a standard digestion method for comparison. Under the optimum conditions, with deposition time of 180 s, linear calibration graphs in range of 10-300 MUg L(-1) Zn(II), 5 200 MUg L(-1) Cd(II), 10-200 MUg L(-1) Pb(II), 20-400 MUg L(-1) Cu(II) were obtained with detection limit of 3.6, 0.1, 0.7 and 4.3 MUg L(-1), respectively. Relative standard deviation were 4.2, 2.6, 3.1 and 4.7% for seven replicate analyses of 27 MUg L(-1) Zn(II), 13 MUg L(-1) Cd(II), 13 MUg L(-1) Pb(II) and 27 MUg L(-1) Cu(II), respectively. The system was validated by certified reference material of trace metals in natural water (SRM 1640 NIST). The developed system was successfully applied for speciation of Cd(II) Pb(II) and Cu(II) in ground water samples collected from nearby zinc mining area. PMID- 21641454 TI - Repeatability in column preparation of a reversed-phase C18 monolith and its application to separation of tocopherol homologues. AB - This work investigated the repeatability of column preparation for a reversed phase C18 monolith, namely stearyl methacrylate-co-ethylene glycol dimethacrylate (SMA-EDMA). The columns were thermally polymerised using three commonly available heating devices (GC oven, hot air oven and water bath) and their chromatographic performance evaluated using micro-liquid chromatography for separation of five test compounds. Precision in terms of %RSD of retention times were 9.0, 6.5, and 12.5 using GC oven, hot air oven and water bath, respectively. Between-batch precision for the hot air oven (n=3 days) was less than 10.4% for retention time. The SMA-EDMA monolith was applied to the separation of tocopherol homologues by capillary electrochromatography. Usually tocopherol homologues cannot be completely separated by conventional reversed-phase C8- or C18-packed bed or C18 silica based monolithic columns. Polymer monolith has been shown to give remarkable selectivity towards the tocopherols compared to the conventional microparticulate phase and silica based monolith. Successful separation of the tocopherol isomers was achieved on the SMA-EDMA monolith without any column modification. PMID- 21641455 TI - An automatic system for acidity determination based on sequential injection titration and the monosegmented flow approach. AB - An automatic sequential injection system, combining monosegmented flow analysis, sequential injection analysis and sequential injection titration is proposed for acidity determination. The system enables controllable sample dilution and generation of standards of required concentration in a monosegmented sequential injection manner, sequential injection titration of the prepared solutions, data collecting, and handling. It has been tested on spectrophotometric determination of acetic, citric and phosphoric acids with sodium hydroxide used as a titrant and phenolphthalein or thymolphthalein (in the case of phosphoric acid determination) as indicators. Accuracy better than |4.4|% (RE) and repeatability better than 2.9% (RSD) have been obtained. It has been applied to the determination of total acidity in vinegars and various soft drinks. The system provides low sample (less than 0.3 mL) consumption. On average, analysis of a sample takes several minutes. PMID- 21641456 TI - Electrochemical detection on electrowetting-on-dielectric digital microfluidic chip. AB - In this work, the use of three-electrode electrochemical sensing system with an electrowetting-on-dielectric (EWOD) digital microfluidic device is reported for quantitative analysis of iodide. T-junction EWOD mixer device was designed using arrays of 50-MUm spaced square electrodes for mixing buffer reagent and analyte droplets. For fabrication of EWOD chips, 5-MUm thick silver EWOD electrodes were formed on a glass substrate by means of sputtering and lift-off process. PDMS and Teflon thin films were then coated on the electrodes by spin coating to yield hydrophobic surface. An external three-electrode system consisting of Au working, Ag reference and Pt auxiliary wires were installed over EWOD electrodes at the end of T-junction mixer. In experiment, a few-microliter droplets of Tris buffer and iodide solutions were moved toward the mixing junction and transported toward electrochemical electrodes by EWOD process. A short processing time within seconds was achieved at EWOD applied voltage of 300V. The analyte droplets mixed with different concentrations were successfully analyzed by cyclic voltametry. Therefore, the combination of EWOD digital microfluidic and electrochemical sensing system has successfully been demonstrated for rapid chemical analysis with minimal reagent consumption. PMID- 21641457 TI - AAO-CNTs electrode on microfluidic flow injection system for rapid iodide sensing. AB - In this work, carbon nanotubes (CNTs) nanoarrays in anodized aluminum oxide (AAO CNTs) nanopore is integrated on a microfluidic flow injection system for in channel electrochemical detection of iodide. The device was fabricated from PDMS (polydimethylsiloxane) microchannel bonded on glass substrates that contains three-electrode electrochemical system, including AAO-CNTs as a working electrode, silver as a reference electrode and platinum as an auxiliary electrode. Aluminum, stainless steel catalyst, silver and platinum layers were sputtered on the glass substrate through shadow masks. Aluminum layer was then anodized by two-step anodization process to form nanopore template. CNTs were then grown in AAO template by thermal chemical vapor deposition. The amperometric detection of iodide was performed in 500-MUm-wide and 100-MUm-deep microchannels on the microfluidic chip. The influences of flow rate, injection volume and detection potential on the current response were optimized. From experimental results, AAO-CNTs electrode on chip offers higher sensitivity and wider dynamic range than CNTs electrode with no AAO template. PMID- 21641458 TI - Exploiting green analytical procedures for acidity and iron assays employing flow analysis with simple natural reagent extracts. AB - Green analytical methods employing flow analysis with simple natural reagent extracts have been exploited. Various formats of flow based analysis systems including a single line FIA, a simple lab on chip with webcam camera detector, and a newly developed simple lab on chip system with reflective absorption detection and the simple extracts from some available local plants including butterfly pea flower, orchid flower, and beet root were investigated and shown to be useful as alternative self indicator reagents for acidity assay. Various tea drinks were explored to be used for chromogenic reagents in iron determination. The benefit of a flow based system, which allows standards and samples to go through the analysis process in exactly the same conditions, makes it possible to employ simple natural extracts with minimal or no pretreatment or purification. The combinations of non-synthetic natural reagents with minimal processed extracts and the low volume requirement flow based systems create some unique green chemical analyses. PMID- 21641459 TI - Sequential injection spectrophotometric determination of tetracycline antibiotics in pharmaceutical preparations and their residues in honey and milk samples using yttrium (III) and cationic surfactant. AB - A sequential injection analysis (SIA) spectrophotometric method for determining tetracycline (TC), chlortetracycline (CTC) and oxytetracycline (OTC) in different sample matrices were described. The method was based on the reaction between tetracyclines and yttrium (III) in weak basic micellar medium, yielding the light yellow complexes, which were monitored at 390, 392 and 395 nm, respectively. A cationic surfactant, cetyltrimethylammonium bromide (CTAB) was used to obtain the micellar system. The linear ranges of calibration graphs were between 1.0 * 10( 5) and 4 * 10(-4) mol L(-1), respectively. The molar absorptivities were 5.24 * 10(5), 4.98 * 10(4) and 4.78 * 10(4) L mol(-1)cm(-1). The detection limits (3sigma) were between 4.9 * 10(-6) and 7.8 * 10(-6) mol L(-1) whereas the limit of quantitations (10sigma) were between 1.63 * 10(-5) and 2.60 * 10(-5) mol L(-1) the interday and intraday precisions within a weak revealed as the relative standard deviations (R.S.D., n=11) were less than 4%. The method was rapid with a sampling rate of over 60 samples h(-1) for the three drugs. The proposed method has been satisfactorily applied for the determination of tetracycline and its derivatives in pharmaceutical preparations together with their residues in milk and honey samples collected in Chiang Mai Province. The accuracy was found to be high as the Student's t-values were found to be less than the theoretical ones. The results were compared favorably with those obtained by the conventional spectrophotometric method. PMID- 21641460 TI - Influenza epidemics: do we really want more protection? PMID- 21641461 TI - Hope for health in Nigeria. PMID- 21641462 TI - Ensuring HIV treatment for all in the USA. PMID- 21641463 TI - Specialist nurses and lung cancer. PMID- 21641464 TI - Oxford University should stop investing in arms companies. PMID- 21641465 TI - A blueprint for country-driven development. PMID- 21641467 TI - David Eagleman. PMID- 21641468 TI - Doctor. PMID- 21641469 TI - Lomita Bekele: delivering health care to an Ethiopian village. PMID- 21641470 TI - Global trends in body-mass index. PMID- 21641471 TI - Open letter to UK Prime Minister David Cameron and Health Secretary Andrew Lansley on safety of medicines. PMID- 21641472 TI - Global trends in body-mass index. PMID- 21641474 TI - Global trends in body-mass index. PMID- 21641475 TI - C-reactive protein in the Heart Protection Study. PMID- 21641478 TI - Eltrombopag for chronic immune thrombocytopenia. PMID- 21641479 TI - Improvement of air quality in low-income countries. PMID- 21641480 TI - The great pretender. PMID- 21641481 TI - The need for technologies to prevent bilirubin-induced neurologic dysfunction syndrome. AB - Dramatic improvements in the overall socioeconomic conditions have yet to impact the unacceptably high maternal (approximately 1500 maternal deaths daily, worldwide) and neonatal morbidity and mortality (more than 10,000 deaths per daily 200,000 live-births, worldwide) in the developing nations. Thus, nations with emerging markets have unique health-societal needs. All infants require a safer transition from a birthing facility to home during the first week after birth and providing for a nurturing environment to prevent neonatal illnesses is integral to "good clinical practice." The unmonitored occurrence of severe hyperbilirubinemia and kernicterus are emblematic of a fractured maternal child healthcare system. The "know-do" gaps that span private versus public health care systems in the emerging markets have led us to conclude that building an interdisciplinary leadership approach to provide innovative strategies and affordable technologies will help bridge and access existing social barriers in the micro- and macro-health environments. Thus, unfettered access, global benchmarks, and culturally relevant strategies are dependent on evidence-based affordable technologies to successfully transform societal health care practices. Implementation of jaundice-related technologies should serve as a template for other affordable newborn health products. PMID- 21641482 TI - The clinical syndrome of bilirubin-induced neurologic dysfunction. AB - We believe that the syndrome of bilirubin-induced neurologic dysfunction [BIND] represents a spectrum of neurologic manifestations among vulnerable infants who have experienced an exposure to bilirubin of lesser degree than generally described in previous publications. Clinical neuro-motor manifestations extend to a range of subtle processing disorders with objective disturbances of visual motor, auditory, speech, cognition, and language among infants with a previous history of moderate-to-severe hyperbilirubinemia of varied duration. Confounding effects include prematurity, hemolysis, perinatal-neonatal complications, altered bilirubin-albumin binding, severity and duration of bilirubin exposure, and the individual vulnerability of the infant related to genetic, family, social, and educational predilection, regardless of the cause of neonatal jaundice. Tools to better assess BIND specific domains of multisensory processing disorders, identified by pyschometric, audiologic, speech, language and visual-motor, and neuromotor examination would allow for prospective surveillance of infants at risk for the syndrome. PMID- 21641483 TI - Bilirubin injury to neurons and glial cells: new players, novel targets, and newer insights. AB - Encephalopathy by hyperbilirubinemia in infants has been described for decades, but neither the underlying cellular and molecular mechanisms nor the selective pattern of bilirubin deposition in the brain is well understood. The brain is composed of highly specialized and diverse populations of cells, represented by neurons and glia that comprise astrocytes, oligodendrocytes, and microglia. Although microscopic evaluation of icteric brain sections revealed bilirubin within neurons, neuronal processes, and microglia, cell dependent-sensitivity to bilirubin toxicity and the role of each nerve cell type are poorly understood. Even less considered are glial and neuronal pathologic alterations as integrated phenomena. The available knowledge on reactivity of glial cells to bilirubin and on the impairment to neuronal network dynamics that it causes, here summarized, suggests that a better comprehension of the interplay between neurons and glia is essential to understand bilirubin neurotoxicity and highlight potential molecular targets that may lead to disease-modifying therapeutic approaches. PMID- 21641484 TI - Bilirubin production and the risk of bilirubin neurotoxicity. AB - Neonatal jaundice usually occurs in the transitional period after birth, presenting as an elevation of circulating bilirubin. Bilirubin neurotoxicity can occur if the levels of bilirubin become excessive (hyperbilirubinemia). This pathologic phenotype of newborn jaundice can develop because of excessive bilirubin production or impaired conjugation, with the risk for developing bilirubin-induced neurologic dysfunction, depending on the degree of the resultant bilirubin load. The plasma bilirubin level thus can be used to assess an infant's risk for developing bilirubin neurotoxicity relative to an infant's age in hours. Because all infants have an impaired conjugation ability, infants at greatest risk are those who have increased bilirubin production rates, because of hemolysis, for example. Therefore, developing potential preventive strategies as well as noninvasive technologies to treat and to identify infants with increased bilirubin production rates, respectively, are tantamount to reducing the incidence of bilirubin-induced neurologic dysfunction. PMID- 21641485 TI - Assessment of UGT polymorphisms and neonatal jaundice. AB - Elevation of the serum bilirubin level is a common, if not universal, finding during the first week of life. This can be a transient phenomenon that resolves spontaneously or can signify a serious or even life-threatening condition. There are many causes of hyperbilirubinemia and related therapeutic and prognostic implications. The diseases in which there is a primary disorder of the metabolism of bilirubin will be reviewed regarding their clinical presentation, pathophysiology, diagnosis, and treatment. These disorders-Gilbert's syndrome and Crigler-Najjar Syndrome-both involve abnormalities in bilirubin conjugation secondary to deficiency of bilirubin uridine diphosphate glucuronosyltransferase. The purpose of this article is to review the current understanding of the genetic polymorphisms that result in these diseases and discuss recent advances in diagnosis and treatment. PMID- 21641487 TI - The status of bilirubin measurements in U.S. laboratories: why is accuracy elusive? AB - In 2003, the Chemistry Resource Committee of the College of American Pathologists introduced a new specimen in the Neonatal Bilirubin Survey. The specimen, consisting of human serum enriched with unconjugated bilirubin and thus resembling a clinical specimen, brought about an improvement in the accuracy of the measurement of bilirubin by laboratories participating in the Neonatal Bilirubin Survey. There was also an improvement in the specificity of methods measuring direct bilirubin. However, persisting inaccuracies and variability in laboratory performance have been traced to calibrators consisting of bovine serum spiked with unconjugated bilirubin and ditaurobilirubin; bovine serum causes underestimation of both bilirubins by 8 major chemical analyzers. To eradicate inaccuracy calibrators and Survey specimens should be made in human instead of bovine serum. PMID- 21641486 TI - Newborn jaundice technologies: unbound bilirubin and bilirubin binding capacity in neonates. AB - Neonatal jaundice (hyperbilirubinemia), which is extremely common in neonates, can be associated with neurotoxicity. A safe level of bilirubin has not been defined in either premature or term infants. Emerging evidence suggest that the level of unbound (or "free") bilirubin has a better sensitivity and specificity than total serum bilirubin for bilirubin-induced neurotoxicity. Although recent studies suggest the usefulness of free bilirubin measurements in managing high risk neonates, including premature infants, no widely available method exists to assay the serum free bilirubin concentration. To keep pace with the growing demand, in addition to reevaluation of old methods, several promising new methods are being developed for sensitive, accurate, and rapid measurement of free bilirubin and bilirubin binding capacity. These innovative methods need to be validated before adopting for clinical use. We provide an overview of some promising methods for free bilirubin and binding capacity measurements with the goal to enhance research in this area of active interest and apparent need. PMID- 21641488 TI - Diagnostic laboratory technologies for the fetus and neonate with isoimmunization. AB - Maternal-fetal blood group incompatibility is common but less commonly results in hemolytic disease of the fetus and newborn (HDFN). HDFN is associated with greater peak bilirubin, at an earlier age, and for longer duration than other causes of hyperbilirubinemia. It poses a substantial risk for kernicterus and accounts for the majority of exchange transfusions for hyperbilirubinemia. Advances in diagnosis and management are described, from identification of the alloimmunized pregnancy by maternal ABO and Rh typing, antibody screen (indirect Coombs test), identification and titration; laboratory evaluation of the maternal fetal unit with a critical maternal antibody titer to prompt fetal antigen status determination; assessment of fetomaternal hemorrhage by conventional Kleihauer Betke testing or by flow cytometric methodology; to antenatal management of isoimmunization and fetal status assessments using the systems of Liley, Queenan, and serial Doppler fetal middle cerebral artery peak velocity measurements. The utility of laboratory diagnostics in the approach to hemolysis in the neonate, including hematology, chemistry, and peripheral blood smear review, is reviewed. The goal of management, to deliver a healthy infant at or near term, is attained for the majority of cases using current modalities; future directions include noninvasive genotyping of fetal blood from maternal serum to fully eliminate RhD alloimmunization and HDFN; and development of prophylaxis and intervention strategies for non-RhD alloimmunizations for which immune globulin is currently unavailable. PMID- 21641489 TI - Neonatal screening for glucose-6-phosphate dehydrogenase deficiency: biochemical versus genetic technologies. AB - Glucose-6-phosphate dehydrogenase (G-6-PD) deficiency, a commonly occurring genetic condition, is associated in neonates with severe hemolytic episodes, extreme hyperbilirubinemia, and bilirubin encephalopathy. Neonatal screening programs for the condition should increase parental and caretaker awareness, thereby facilitating early access to treatment with resultant diminished mortality and morbidity. However, screening for G-6-PD deficiency is not widely performed. Although G-6-PD-deficient males may be accurately identified, females are more difficult to categorize because many in this group may be heterozygotes with phenotype overlap between normal homozygotes, heterozygotes, and deficient homozygotes. Screening methodologies include biochemical qualitative assays, quantitative enzymatic activity measurements and DNA-based polymerase chain reaction molecular screening. The appropriateness of any of these technologies for any particular population group or geographic area must be assessed before setting up a screening program. The pros and cons of each method, including ease of testing, cost, need for sophisticated laboratory equipment and degree of personnel training, as well as the ability to identify females, are discussed. PMID- 21641490 TI - Auditory impairment in infants at risk for bilirubin-induced neurologic dysfunction. AB - Classical and subtypes of kernicterus associated with bilirubin toxicity can be differentiated in part with physiological auditory measures that include auditory evoked potentials and measures of cochlear integrity. The combination of these auditory measures suggests that bilirubin exposure results in auditory system damage initially at the level of the brainstem, progressing to the level of the VIII cranial nerve and then to greater neural centers. There is no evidence of neural damage at the level of the cochlea. Auditory neural damage from bilirubin toxicity ranges from neural timing deficits, including neural firing delays and dyssynchrony, to neural response reduction and even elimination of auditory neural responses. This condition is comprehensively described as auditory neuropathy spectrum disorder. Independent measures of cochlear function and auditory neural function up to the level of the brainstem can effectively diagnose auditory neural damage resulting from bilirubin neurotoxicity. Intervention, including cochlear implants can be effective. PMID- 21641491 TI - The role of phototherapy in the crash-cart approach to extreme neonatal jaundice. AB - Extreme neonatal jaundice occurs infrequently but carries a high risk of permanent sequelae (kernicterus) when it does. Rapid therapeutic intervention has the potential to reduce this risk in some infants. Several case reports of infants with acute intermediate to advanced bilirubin encephalopathy shows that reversal may be possible. Phototherapy can be instituted at the flip of a switch, whereas other therapeutic measures necessarily involve delays. Therefore, high intensity phototherapy must be regarded as an emergency measure in infants presenting with extreme jaundice and even more so in the presence of neurological symptoms. The principal and well-described effect of phototherapy involves conversion of bilirubin IXalpha (z, z) to more polar isomers, which are excreted in bile and urine. When care is taken to maximize the spectral power of phototherapy lights, and whenever possible with measures added to reduce the enterohepatic circulation of bilirubin, very rapid reductions in total serum bilirubin levels are possible. A hypothesis has been advanced that conversion of bilirubin to more polar photoisomers, which can reach relative concentrations of 20%-25% of total serum bilirubin within 1-2 hours, might have a direct neuroprotective effect. This theory posits that because polar molecules generally require a transporter to cross the blood-brain barrier, bilirubin photoisomers should be less prone to enter the brain. Although this theory has some support in in vitro toxicity studies, the evidence is controversial. Until further experimental support can be gained, photoconversion of bilirubin does not constitute a viable argument against instituting further measures against bilirubin neurotoxicity, such as intravenous immune globulin (when indicated) and exchange transfusion. Conversely, neither is the state of evidence an argument against immediate and effective phototherapy in the medical emergency of extreme neonatal jaundice. PMID- 21641492 TI - Blood exchange transfusion for infants with severe neonatal hyperbilirubinemia. AB - Blood exchange transfusion has become a rare event in most developed countries. As a result, many pediatricians may not have performed or even seen one. However, it remains a frequent emergency rescue procedure for severe neonatal hyperbilirubinemia in many underdeveloped regions of the world. Conventionally, exchange transfusion has been performed via a central umbilical venous catheter by pull-push cycle method and recently peripheral artery/peripheral vein has emerged as an alternative, isovolumetric route. Continuous arterio-venous exchange is possibly more effective though its automation has not been successful. Concerns for procedural and operator related adverse events have been raised in the context of declining indications. A required continued expertise for this life-saving intervention, in the face of rare but critical hyperbilirubinemia and/or unrecognized hemolytic diseases, deserves adaptation of newer technologies to make neonatal exchange transfusion a safer and more effective procedure. Technological innovations and simulation technologies are urgently needed. PMID- 21641493 TI - A global need for affordable neonatal jaundice technologies. AB - Globally, health care providers worldwide recognize that severe neonatal jaundice is a "silent" cause of significant neonatal morbidity and mortality. Untreated neonatal jaundice can lead to death in the neonatal period and to kernicterus, a major cause of neurologic disability (choreo-athetoid cerebral palsy, deafness, language difficulty) in children who survive this largely preventable neonatal tragedy. Appropriate technologies are urgently needed. These include tools to promote and enhance visual assessment of the degree of jaundice, such as simpler transcutaneous bilirubin measurements and readily available serum bilirubin measurements that could be incorporated into routine treatment and follow-up. Widespread screening for glucose-6-phoshate dehydrogenase deficiency is needed because this is often a major cause of neonatal jaundice and kernicterus worldwide. Recognition and treatment of Rh hemolytic disease, another known preventable cause of kernicterus, is critical. In addition, effective phototherapy is crucial if we are to make kernicterus a "never-event." Finally it is essential that we conduct appropriate population-based studies to accurately elucidate the magnitude of the problem. However, knowledge alone is not sufficient. If we are to implement these and other programs and technologies to relegate severe neonatal jaundice and its sequelae to the history books, screening and interventions must be low cost and technologically appropriate for low and middle income nations. PMID- 21641494 TI - The need to implement effective phototherapy in resource-constrained settings. AB - Phototherapy is the treatment of choice to reduce the severity of neonatal unconjugated hyperbilirubinemia regardless of its etiology. Its implementation requires a technical framework that conforms to existing evidence-based guidelines that promote its safer and effective use worldwide. Optimal use of phototherapy has been defined by specific ranges of total serum bilirubin thresholds configured to an infant's postnatal age (in hours) and potential risk for bilirubin neurotoxicity. Effective phototherapy implies its use at specific blue light wavelengths (peak emission, 450 +/- 20 nm) and emission spectrum (range, 400-520 nm), preferably in a narrow bandwidth that is delivered at an irradiance of >=30 MUW/cm(2)/nm to up to 80% of an infant's body surface area. However, this is often not feasible in clinical settings with limited or constrained resources. To identify and bridge implementation barriers, we propose minimum criteria for device performance for safe and practical use of phototherapy as a prophylactic intervention to prevent severe hyperbilirubinemia. PMID- 21641495 TI - Preface to mitochondria and cardioprotection. PMID- 21641496 TI - [An editorial note about the editorials in the Gaceta Sanitaria]. PMID- 21641498 TI - Failure of the afferent limb: a persistent problem in rapid response systems. PMID- 21641499 TI - Rescue and resuscitation or body retrieval--the dilemmas of search and rescue efforts in drowning incidents. PMID- 21641501 TI - Pneumonia in the long-term resident. AB - Pneumonia in the long-term resident is common. It is associated with high morbidity and mortality. However, diagnosis and management of pneumonia in long term care residents is challenging. This article provides an overview of the epidemiology, pathophysiology, diagnostic challenges, and management recommendations for pneumonia in this setting. PMID- 21641502 TI - Managing the patient with dementia in long-term care. AB - The majority of residents in a nursing home have some degree of dementia. The prevalence is commonly from 70% to 80% of residents. This article covers the following topics on caring for patients with dementia in long-term care: (1) the efficacy of cholinesterase inhibitors and memantine, (2) the optimal environment for maintenance of function in moderate dementia, (3) the treatment of depression and agitation, and (4) the evaluation and management of eating problems. PMID- 21641503 TI - Palliative care for patients with dementia in long-term care. AB - Seventy percent of people in the United States who have dementia die in the nursing home. This article addresses the following topics on palliative care for patients with dementia in long-term care: (1) transitions of care, (2) infections, other comorbidities, and decisions on hospitalization, (3) prognostication, (4) the evidence for and against tube feeding, (5) discussing goals of care with families/surrogate decision makers, (6) types of palliative care programs, (7) pain assessment and management, and (8) optimizing function and quality of life for residents with advanced dementia. PMID- 21641504 TI - Medications in long-term care: when less is more. AB - Attention has been drawn to the potential risks of several medications in the long-term care setting. Most of these medications deemed as inappropriate affect the central nervous system and are indicated only for select populations with specific conditions. Many of these drugs are prescribed without clear indications and continued indefinitely without critical decision-making about the potentially salutary effects of discontinuing medications. This article describes the increasing awareness of potentially inappropriate prescribing in the long-term care setting and reviews the rationale for why various types of medications are deemed inappropriate, with a focus on agents that affect central nervous system functioning. PMID- 21641505 TI - Evidence-based medicine (EBM): what long-term care providers need to know. AB - Evidence-based medicine (EBM) has been widely used in medicine for 2 decades. Recently, EBM has become a central part of reforming nursing homes and quality improvement. It is very important for long-term care providers to practice EBM. This article briefly introduces the concept of EBM; addresses some potential benefits, harms, and challenges of practicing EBM in long-term care settings; and promotes EBM and its appropriate use among long-term care providers. PMID- 21641506 TI - Update on teaching in the long-term care setting. AB - Since the advent of the teaching nursing home, made formal in the 1980s, long term care has been used to teach geriatric medicine. Despite this, national surveys have indicated a need for more training during residency to facilitate the appropriate care for the frail long-term care patient population. In addition to medical knowledge, the long-term care site is appropriate for teaching the Accreditation Council of Graduate Medical Education's core competencies of "practice-based learning and improvement," "interpersonal and communication skills," and "systems-based practice." Program planners should emphasize opportunities for students to demonstrate their skill in one of these competencies. PMID- 21641507 TI - Nausea and other nonpain symptoms in long-term care. AB - There is a need to improve the quality of end-of-life care in nursing homes by improving the timely assessment and management of various sources of suffering. Much of the research/discussion in this area has focused on the assessment and treatment of pain. This article reviews the frequency and management of nonpain symptoms in the long-term care setting, particularly focusing on patients at the end of life. Although the long-term care setting presents challenges to effective management, an approach for addressing these challenges is discussed and applied to 3 commonly encountered nonpain symptoms. PMID- 21641508 TI - Urinary tract infections in long-term care residents. AB - Urinary tract infection (UTI) is common in long-term care (LTC) residents; however, most infections are asymptomatic and do not require treatment. Differentiating asymptomatic from symptomatic UTI is challenging, because LTC residents typically have chronic genitourinary complaints, multiple comorbid illnesses, and communication barriers. Although consensus guidelines have been proposed to improve the accuracy of identifying symptomatic UTIs and minimize treatment of asymptomatic UTIs, diagnostic accuracy is not yet optimized. Strategies for prevention of UTI are unsatisfactory and require further study; nevertheless, there is some evidence for the efficacy of cranberry products and vaginal estrogen to prevent recurrent UTI in women. PMID- 21641509 TI - Pressure ulcers in long-term care. AB - Pressure ulcers are common, costly, and debilitating chronic wounds, which occur preferentially in people with advanced age, physical or cognitive impairments, and multiple comorbidities. Residents with pressure ulcers have decreased quality of life and increased morbidity and mortality, and facilities with high rates of pressure ulcers have higher costs and risks of litigation. Health professionals who practice in this setting should be well versed in pressure ulcer management. This article reviews the significance, risk factors, pathophysiology, prevention, diagnosis, and management of pressure ulcers in long-term care. PMID- 21641510 TI - Transitional care of the long-term care patient. AB - This article reviews the literature on transitional care to and from the LTC environment, highlighting strategies to improve the quality of care transitions. Several factors are vital in the improvement of systems of care dealing with transitions. Key factors include communication with and among health care providers, effective medication reconciliation, advanced discharge planning, and timely use of palliative care. PMID- 21641511 TI - Doing dementia better: anthropological insights. AB - Dementia, or neurodegenerative disease, is a disease category, and yet it is widely described in popular and professional media as a horror story. Patients with dementia and their families frequently report that they are less than pleased with their clinical encounters. This article reveals the deleterious impact that cultural assumptions about dementia have on the care provided, and, through an exploration of anthropological theories of personhood, suggests strategies for seeking improved quality of life through personhood-centered care. PMID- 21641512 TI - Long-term care of the aging population with intellectual and developmental disabilities. AB - The aging population with intellectual and developmental disabilities (I/DD) deserves appropriate health care and social support. This population poses unique medical and social challenges to the multidisciplinary team that provides care. In the past, long-term care (LTC) facilities played an essential role in the livelihood of this population. The likelihood that the geriatric LTC system must prepare for adequately caring for this population is high. This article conveys the need to prepare for the inclusion of the growing aging population with I/DD into long-term care with the general elderly population in the near future. PMID- 21641513 TI - Cancer in long-term care. AB - This article describes the range of cancer patients in longterm care and provides a framework for clinical decision making. The benefits and burdens of providing standard therapy to a vulnerable population are discussed. To give more specific guidelines for advocates of treatment, skeptics, and others, the authors present best estimates of the current burden of cancer in the long-term care population and current screening guidelines that apply to the elderly under long-term care. Experience-based suggestions are offered for oncologists and clinicians involved in long-term care to help them respond to patient and family concerns about limitations of cancer care. PMID- 21641514 TI - A pilot study of yoga treatment in children with functional abdominal pain and irritable bowel syndrome. AB - OBJECTIVES: The aim of this pilot study was to evaluate the effect of yoga exercises on pain frequency and intensity and on quality of life in children with functional abdominal pain. DESIGN: 20 children, aged 8-18 years, with irritable bowel syndrome (IBS) or functional abdominal pain (FAP) were enrolled and received 10 yoga lessons. Pain intensity and pain frequency were scored in a pain diary and quality of life was measured with the Kidscreen quality of life questionnaire (KQoL). RESULTS: In the 8-11 year old group and the 11-18 year old group pain frequency was significantly decreased at the end of therapy (p=0.031 and p=0.004) compared to baseline. In the 8-11 year group pain intensity was also significantly decreased at this time point (p=0.015). After 3 months there still was a significant decrease in pain frequency in the younger patient group (p=0.04) and a borderline significant decrease in pain frequency in the total group (p=0.052). Parents reported a significantly higher KQoL-score after yoga treatment. CONCLUSION: This pilot study suggests that yoga exercises are effective for children aged 8-18 years with FAP, resulting in significant reduction of pain intensity and frequency, especially in children of 8-11 years old. PMID- 21641515 TI - Association between tongue appearance in Traditional Chinese Medicine and effective response in treatment of rheumatoid arthritis. AB - OBJECTIVE: Explore the associations between the tongue appearances in Traditional Chinese Medicine (TCM) and effective response (ACR20 response based on American College of Rheumatology) in rheumatoid arthritis (RA) patients treated with Chinese medicine (CM) and western biomedical combination therapy (WM). METHODS: This study used the data from a previous multi-center randomized-controlled clinical trial. Data pertaining to tongue coating and tongue body color were collected. In order to simplify the tongue diagnosis for easily understood by biomedical professionals, only two typical tongue coating (white and yellow) and four typical tongue body colors (purple, pink, pale and red) were identified for this analysis. 170 cases with clear tongue coating and 198 cases with clear tongue body color in TCM treatment (Glucosidorum Tripterygll Totorum tablets and Yishen Juanbi tablets) group, 181 cases with identified tongue coating and 189 cases with identified tongue body color in WM treatment (diclofenec, methotrexate and sulfasalazine) group were included for the analysis. The ACR20 response at 12 weeks and 24 weeks were used as an outcome measure of efficacy. The effective rates in patients with different tongue appearances were analyzed with Chi-square method and the association between the changes of tongue coating/body color and the ACR20 response was analyzed with a repeated measures logistic regression analysis. RESULTS: At 12 weeks, the ACR20 responses in the patients treated with CM and WM therapy were 33.6% and 53.0%, respectively, and at 24 weeks, they were 57.9% and 84.3%, respectively. RA patients with white tongue coating showed higher effective rate than those patients with yellow tongue coating in the treatment with WM intervention (p<0.05), and there was no difference in the patients with CM intervention. Further association analysis showed that TCM would be less effective for the patients with pale tongue body (p=0.0323), and WM would be less effective for the patients with purple or red tongue body (p=0.0291 and 0.0027, respectively). CONCLUSION: TCM was less effective for the patients with pale tongue body, and WM was be less effective for the patients with purple or red tongue body, or white tongue coating. The results suggest that tongue coating and body color might be used to help identify a subset of RA patients both for CM and WM interventions. PMID- 21641516 TI - Effect of yogic practices on lipid profile and body fat composition in patients of coronary artery disease. AB - OBJECTIVES: To observe the effect of regular yogic practices and self-discipline in reducing body fat and elevated lipids in CAD patients. METHOD: In this study one hundred seventy (170) subjects, of both sexes having coronary artery disease were randomly selected form Department of Cardiology. Subjects were divided in to two groups randomly in yoga group and in non-yoga group, eighty five (85) in each group. Out of these (170 subjects), one hundred fifty four (154) completed the study protocol. TIME LINE: The yogic intervention consisted of 35-40 min/day, five days in a week till six months in the Department of Physiology CSMMU UP Lucknow. Body fat testing and estimation of lipid profile were done of the both groups at zero time and after six months of yogic intervention in yoga group and without yogic intervention in non yoga group. RESULTS: In present study, BMI (p<0.04), fat % (p<0.0002), fat free mass (p<0.04), SBP (p<0.002), DBP (p<0.009), heart rate (p<0.0001), total cholesterol (p<0.0001), triglycerides (p<0.0001), HDL (p<0.0001) and low density lipoprotein (p<0.04) were changed significantly. CONCLUSION: Reduction of SBP, DBP, heart rate, body fat%, total cholesterol, triglycerides and LDL after regular yogic practices is beneficial for cardiac and hypertensive patients. Therefore yogic practices included in this study are helpful for the patients of coronary artery disease. PMID- 21641517 TI - Are complementary medicine practitioners implementing evidence based practice? AB - BACKGROUND: Over the past few decades the health professions have witnessed increasing pressure to shift from a culture of delivering care based on tradition and intuition, to a situation where decisions are guided and justified by the best available evidence. While there are concerns that many complementary and alternative medicine (CAM) practitioners may be cautious about embracing such an approach, no studies to date have effectively tested this assumption. OBJECTIVE: To identify the skills, attitude, training and use of evidence-based practice (EBP) amongst CAM practitioners. DESIGN: Descriptive survey, using the evidence based practice attitude and utilisation survey (EBASE). SUBJECTS: Randomly selected nationwide sample of system-based, non-medically qualified CAM practitioners practicing in a clinical capacity within Australia. MAIN OUTCOME MEASURES: Practitioner skill, attitude, training and use of EBP. RESULTS: Of the 351 questionnaires successfully dispatched, 126 were returned (36%). Most practitioners believed EBP was useful (92%) and necessary (73%) in CAM practice. While the majority of clinicians (>74%) reported participation in EBP activities, albeit infrequently, only a small to moderate proportion of decisions were based on evidence from clinical trials, with most practitioners relying on traditional knowledge, textbooks and clinical practice guidelines. Lack of available evidence, time, industry support and skills were perceived as barriers to EBP uptake. CONCLUSIONS: While the small response rate limits the generalisability of these findings, the sample was considered representative of Australian CAM practitioners. What this study shows is that even though CAM practitioners may be supportive of EBP, education and training is needed to further improve clinician understanding and application of evidence-based practice. PMID- 21641518 TI - Use of Chinese medicine by women with breast cancer: a nationwide cross-sectional study in Taiwan. AB - OBJECTIVES: The National Health Insurance (NHI) provided Western medicine (WM) and Chinese medicine (CM) in Taiwan. This study aims to explore CM use by women with breast cancer under NHI. METHODS: Using NHI Research Database, a retrospective cross-sectional study was conducted. Women with breast cancer were identified by diagnosis codes. Their claims of CM and WM outpatient services of 2007 were analyzed. RESULTS: Among 70,012 female breast cancer patients, the prevalence of insurance covered CM was 35.6%. Among all CM users, 4379 (17.5%) patients visited CM for breast cancer. More than half CM users (67.6%) had less than 6 visits and 16.3% users had more than 12 visits. The majority of CM users (87.6%) also used WM ambulatory services. Private clinics (76.7%) provided the most CM services, followed by private hospitals (16.9%). The most frequently used CM therapies were Chinese herbal medicine (80.5%), followed by acupuncture/traumatology manipulative therapies (22.3%). The average cost of CM was US$17.6 per visit and US$119.7 per user annually. CM users were more likely to be younger than 60 years old, employees, with higher income, and not living in Northern Taiwan. CONCLUSIONS: A significant portion of breast cancer patients used insurance covered CM. Most CM users also used WM. The potential of drug-herb interactions should be concerned. PMID- 21641519 TI - Acupuncture for low back pain: a survey of clinical practice in the UK. AB - OBJECTIVES: Acupuncture is recommended in official UK guidelines for persistent non-specific low back pain and is popular with patients. However, what UK-based acupuncturists actually do in every day clinical practice is poorly documented. We therefore conducted a survey of every-day clinical practice of acupuncture for low back pain in the UK. DESIGN: Cross-sectional postal survey. SETTING: Random samples of 100 acupuncturists from each of the three main UK societies for acupuncturists were sent questionnaires. MAIN OUTCOME MEASURES: A questionnaire designed (and pilot-tested) for this study asked about: training and professional identity, current work situation, and typical approach to treating a patient with low back pain. RESULTS: Completed questionnaires were received from 129 respondents (53% male), representing each society approximately equally. Work situation (e.g. weekly hours practicing acupuncture) differed across societies. Respondents reported needling 2-30 acupuncture points in an average treatment (median=8) and leaving needles in for 0-40 min (median=20 min). A large number of individual points (121) were named as 'typical'. CONCLUSIONS: There is huge variation in how acupuncture is used by UK practitioners to treat people with low back pain. This probably constitutes a difficult situation for patients when selecting an acupuncturist. It may also translate into large variation in clinical outcomes for patients receiving care from different acupuncturists as well as difficulty in developing acupuncture protocols for pragmatic and other trials. Such diversity reflects the ongoing debate within the acupuncture profession about good practice for common conditions. PMID- 21641520 TI - Complementary and alternative medicine: perception and use by physiotherapists in the management of low back pain. AB - BACKGROUND AND PURPOSE: The aim of this survey was to investigate complementary and alternative medicine (CAM) use by physiotherapists for the treatment of low back pain (LBP). METHODS: Questionnaires (n=1000) were distributed to physiotherapists randomly selected from the UK's Organisation of Chartered Physiotherapists in Private Practice (OCPPP). Results were analysed by SPSS. RESULTS: A response rate of 46% was obtained (n=459). 94.3% of respondents stated that they currently treated LBP; the most common CAMs used by physiotherapists were acupuncture (46.9%) and massage (2.1%). Physiotherapists perceived acupuncture, massage, osteopathy, chiropractic and yoga to be effective in the treatment of LBP, but were unsure about the effectiveness of other CAMs. DISCUSSION AND CONCLUSION: Physiotherapists in this sample often use acupuncture and massage to treat LBP, but appear to have little knowledge of other CAMs. Physiotherapists use mainstream methods such as mobilization and massage which could be perceived as CAM depending on the physiotherapists' perceptions and training, as these therapies have also been used and investigated by CAM practitioners. More research is required on the use of CAM as little is known regarding the effectiveness of these therapies on LBP. PMID- 21641521 TI - The quality of information on websites selling St. John's wort. AB - BACKGROUND: Health consumers are increasingly using the Internet to access information about health care, to self-diagnose, and to purchase medication. The use of the Internet to purchase herbal products is of particular interest because of the high level of consumer expenditure on herbal medicines, and the misperception by some consumers that herbal products are natural, and thus absent of any contraindications, drug interactions and adverse effects. It is possible that consumers may purchase herbal medicines via the Internet without consulting health professionals and therefore, use these medicines in an unsafe manner. OBJECTIVES: To examine the quality of e-commerce websites that sell herbal products; specifically, websites where St. John's wort (Hypericum perforatum) can be purchased. DESIGN: Cross-sectional survey of 54 selected websites, including online pharmacies, online health food stores and manufacturers of herbal medicines. MAIN OUTCOME MEASURE: A modified version of the DISCERN instrument was used to assess the quality of websites. RESULTS: The majority of websites rated poorly with a concerning lack of information about the interaction between hypericum and warfarin, anti-depressants and oral contraceptives. Most sites also failed to provide sufficient information about the contraindications and adverse effects of hypericum treatment. CONCLUSIONS: The results of this study strongly support the need for improved consumer education about herbal medicine, as well as the application of more stringent standards to websites that sell medications. PMID- 21641522 TI - Effectiveness of light therapy for depression among active duty service members: A nonrandomized controlled pilot trial. AB - OBJECTIVE: The authors investigated the potential effectiveness of light therapy as an augmentation treatment for depression among active duty service members. DESIGN: This pilot study recruited active duty service members deployed to an area of combat operations. Enrollment was offered to service members scoring 50 or greater on the Zung Self-Rating Depression Scale. The authors implemented a systematic sampling technique randomly assigning the first subject and then alternating each subsequent subject to either a reference group which received the usual standard of care plus light therapy at 10,000 lux or a control group which received the usual standard of care and light therapy at 50 lux. Both groups received 90 min light sessions for five days. SETTING: The study was conducted at Walter Reed Army Medical Center's Psychiatry Continuity Service. MAIN OUTCOME MEASURE: Zung Self-Rating Depression Scale collected at baseline, after five consecutive daily light sessions, and one week later. RESULTS: A repeated measures analysis of variance (RM ANOVA) was conducted to examine the change in Zung Depression results which showed a significant main effect for time F(2, 21)=5.05, p<0.02, indicating that depression scores reduced over time for both participant groups. Post hoc comparisons (with Bonferroni correction) demonstrated that the post-treatment Zung score was significantly lower indicating less depression than the baseline Zung score (p<0.004) and there was a statistical trend (p<0.05) for depression scores to be reduced halfway through the study in the treatment group. CONCLUSION: The post hoc analysis hints at the possibility of a reduction in depression during the active phase of light treatment. PMID- 21641523 TI - The effect of extremely diluted agitated gibberellic acid (10e-30) on wheat stalk growth--a two researcher pilot study. AB - OBJECTIVE: Use of a wheat growth bio assay after 7 days in research on homeopathic dilutions of gibberellic acid. METHODS: Grains of winter wheat (Triticum aestivum, Capo variety) were observed under the influence of extremely diluted gibberellic acid (10(-30)) prepared by stepwise dilution and agitation according to a protocol derived from homeopathy (30*). Analogously prepared water was used for control. In a two centre study, 3 experiments with a total of 4880 grains were performed. RESULTS: Data were found to be rather homogeneous within the control group as well as within the verum group in general. Germination rates were around 95%, with no significant difference between verum and control group (p>0.05). Mean stalk lengths (mm) were 40.63+/-20.96 for the verum and 44.33+/ 21.11 for the control group (mean+/-S.D.) at grain level (N=2440 per group) and +/-5.33 and +/-5.89, respectively at dish level (122 cohorts of 20 grains per treatment group). In other words, verum stalk length (91.65%) was 8.35% smaller than control stalk length (100%). This difference is statistically highly significant (p<0.001) and was found by both researchers involved independently. CONCLUSION: These results suggest that there was an influence of gibberellic acid 30* on wheat seedling development, i.e. the wheat growth bio assay can be a useful tool for further experiments on homeopathic dilutions of gibberellic acid. PMID- 21641524 TI - Potential health benefits of simulated laughter: a narrative review of the literature and recommendations for future research. AB - INTRODUCTION: Scientific research has shown that laughter may have both preventive and therapeutic values. Health-related benefits of laughter are mainly reported from spontaneous laughter interventional studies. While the human mind can make a distinction between simulated and spontaneous laughter, the human body cannot. Either way health-related outcomes are deemed to be produced. Simulated laughter is thus a relatively under-researched treatment modality with potential health benefits. The aim of this review was firstly to identify, critically evaluate and summarize the laughter literature; secondly to assess to which extent simulated laughter health-related benefits are currently sustained by empirical evidence; and lastly to provide recommendations and future directions for further research. METHODS: A comprehensive laughter literature search was performed. A list of inclusion and exclusion criteria was identified. Thematic analysis was applied to summarize laughter health-related outcomes, relationships, and general robustness. RESULTS: Laughter has shown different physiological and psychological benefits. Adverse effects are very limited and laughter is practically lacking in counter-indications. Despite the limited number of publications, there is some evidence to suggest that simulated laughter has also some effects on certain aspects of health, though further well-designed research is warranted. CONCLUSIONS: Simulated laughter techniques can be easily implemented in traditional clinical settings for health and patient care. Their effective use for therapeutic purposes needs to be learned, practiced, and developed as any other medical strategy. Practical guidelines and further research are needed to help health care professionals (and others) implement laughter techniques in their health care portfolio. PMID- 21641525 TI - Endothelin: from bench to bedside and back. PMID- 21641526 TI - Modeling the dynamic association of BMI and mortality in the Framingham Heart Study. AB - PURPOSE: To examine and model the dynamic association of BMI and mortality in the Framingham Heart Study (FHS). METHODS: BMI is transformed to facilitate modeling curvature associations. Logistic models are used to demonstrate whether different conclusions may be obtained for the same group of individuals under different settings created from FHS. Time-dependent covariates Cox models are used to model the association of BMI and mortality when the proportional hazards assumptions for Cox models are violated. RESULTS: Both the measurement time of BMI and the length of follow-up affect the conclusions obtained from logistic models, especially for men. Time-dependent covariates Cox models show that the association between BMI and mortality for men depends on the follow-up time, while that for women depends on the age of BMI measurement. CONCLUSION: The association of BMI and mortality in FHS is a dynamic system that traditional analyses methods may lead to different conclusions for different study designs. This finding is consistent with the results of several other studies done from different perspectives, suggesting that the dynamic features demonstrated in FHS may apply to other populations. Advanced methods such as time-dependent covariates Cox models may be helpful for future analysis. PMID- 21641527 TI - Measurement: a question of accuracy. PMID- 21641528 TI - Postanesthesia patients with large upper arm circumference: is use of an "extra long" adult cuff or forearm cuff placement accurate? AB - The purpose of this study was to determine if blood pressure (BP) measured in the forearm or with an extra-long BP cuff in the upper arm accurately reflects BP measured in the upper arm with an appropriately sized BP cuff in patients with large upper arm circumference. A method-comparison design was used with a convenience sample of 49 PACU patients. Noninvasive blood pressures were obtained in two different locations (forearm; upper arm) and in the upper arm with an extra-long adult and recommended large adult cuff sizes. Data were analyzed by calculating bias and precision for the BP cuff size and location and Student's t tests, with P < .0125 considered significant. Significantly higher forearm systolic (P < .0001) and diastolic (P < .0002) BP measurements were found compared to BP obtained in the upper arm with the reference standard BP cuff. Significantly higher systolic (t(48df) = 5.38, P < .0001), but not diastolic (t(48df) = 4.11, P < .019), BP differences were found for BP measured with the extra-long cuff at the upper arm site compared to the upper arm, reference standard BP. Findings suggest that the clinical practice of using the forearm or an extra-long cuff in the upper arm for BP measurement in post anesthesia patients with large upper arm circumferences may result in inaccurate BP values. PMID- 21641529 TI - Agreement between temporal artery, oral, and axillary temperature measurements in the perioperative period. AB - This study examined agreement in temperature readings preoperatively and postoperatively between temporal artery and electronic oral/axillary thermometers as well as the seconds required to obtain temperature readings across the three measuring modes. Using a repeated measures design, 86 adult subjects had temporal artery, oral, and axillary temperatures taken upon admission to the surgical area and upon admission to the PACU. Findings indicated best agreement both preoperatively and postoperatively between the oral mode of the electronic thermometer and the temporal artery thermometer, followed by agreement between oral and axillary modes of the electronic thermometer, and the least agreement between the temporal artery and axillary readings. Seconds to temperature measurement was significantly different both preoperatively and postoperatively, with temporal artery measurement the fastest, followed by oral and then axillary. Results support the use of the temporal artery thermometers as an alternative for perioperative noninvasive temperature monitoring. PMID- 21641530 TI - Esophageal, tympanic, rectal, and skin temperatures in children undergoing surgery with general anesthesia. AB - The purpose of this study was to determine the degrees of agreement between various sites of temperature measurement and examine the trend of body temperature in children during surgery under general anaesthesia. Thirty-six consecutive children who underwent surgery with general anaesthesia, had temperatures measured at the oesophagus, skin, ear canal and rectum at baseline, every 15 minutes for the first hour and every 30 minutes thereafter. Spearman correlation and Bland-Altman analyses were used to compare data and trends of mean differences assessed by line graphs. The median age of the sample was 48 months. There were 575 temperature measurements taken. The inter-method correlation coefficients was highest for the oesophageal vs rectal (r = 0.96) temperature and lowest for rectal vs skin (r = -0.11) temperature. The lowest mean difference (95% CI) in temperature at commencement of surgery was between the oesophageal and rectal sites, -0.03 degrees C (-0.08, -0.01) while the highest mean difference (95% CI) temperature was between oesophageal and skin sites, 3.24 degrees C (2.65, 3.85). The trend in differential temperatures between sites remained throughout the duration of surgery. Bland-Altman plots showed that the least difference (bias) at baseline (0.3 degrees C) was between the oesophageal and tympanic temperatures while at 1 hour (0.13 degrees C ) was between the oesophageal and rectal temperatures. The oesophageal site was the closest to rectal for monitoring core temperature while the skin was the least reliable site in the study population. In the situation where oesophageal probe is not routine or functioning, rectal or tympanic temperatures may be used. PMID- 21641531 TI - Global health care partnerships: a call to perianesthesia nurses. PMID- 21641532 TI - A simple thank you. PMID- 21641533 TI - Intravenous lidocaine for acute pain treatment. PMID- 21641534 TI - Understanding patient safety culture: part I. PMID- 21641535 TI - Obstacles and drawbacks to avoid in qualitative nursing research. PMID- 21641536 TI - Research news: Nurse anesthesia. PMID- 21641537 TI - "I got an 'A', then a lot of red!": Converting an academic paper to a suitable manuscript. PMID- 21641538 TI - A focus on safety: meeting notes from the 2010 ASA Annual Meeting. PMID- 21641539 TI - The normalization of deviance: a threat to patient safety. PMID- 21641540 TI - Feeding mistiming decreases reproductive fitness in flies. AB - The diurnally active fruit flies prefer a major meal in the morning. Feeding the flies in the evening uncouples their metabolic cycle from circadian activity rhythms. A paper by Xu et al. in this issue of Cell Metabolism found that such uncoupled rhythms reduce egg laying. PMID- 21641541 TI - PTPMT1: connecting cardiolipin biosynthesis to mitochondrial function. AB - Cardiolipin, a phospholipid component of the inner mitochondrial membrane, is required for mitochondrial metabolism. In this issue, Zhang et al. (2011) highlight a critical role for PTPMT1, a mitochondrial phosphatase, in cardiolipin biogenesis and possibly in cardiolipin deficiency diseases. Their findings also unveil a yet uncharacterized pathway affecting cell growth. PMID- 21641542 TI - A radical role for TOR in longevity. AB - TOR (target of rapamycin) signaling regulates life span in many organisms, but the mechanism behind the effect is unknown. In this issue of Cell Metabolism, Pan and colleagues (2011) find that reduced TORC1 activity promotes yeast life span via a mechanism that, paradoxically, relies upon the production of normally deleterious reactive oxygen species. PMID- 21641543 TI - Renal nerves, WNK4, glucocorticoids, and salt transport. AB - Roles of the sympathetic nervous system versus kidney salt transporters in hypertension are debated. A study in Nature Medicine (Mu et al., 2011) shows that dietary salt excess, coupled with beta-adrenergic stimulation, increases arterial pressure via glucocorticoid receptors and WNK4, suggesting interactions between these systems in the pathogenesis of hypertension. PMID- 21641544 TI - Fine tuning our cellular factories: sirtuins in mitochondrial biology. AB - Sirtuins have emerged in recent years as critical regulators of metabolism, influencing numerous facets of energy and nutrient homeostasis. Here, we review recent advances on the role of this fascinating family of mammalian proteins and their well-orchestrated function in modulating mitochondrial activity. PMID- 21641545 TI - mRNA expression signatures of human skeletal muscle atrophy identify a natural compound that increases muscle mass. AB - Skeletal muscle atrophy is a common and debilitating condition that lacks a pharmacologic therapy. To develop a potential therapy, we identified 63 mRNAs that were regulated by fasting in both human and mouse muscle, and 29 mRNAs that were regulated by both fasting and spinal cord injury in human muscle. We used these two unbiased mRNA expression signatures of muscle atrophy to query the Connectivity Map, which singled out ursolic acid as a compound whose signature was opposite to those of atrophy-inducing stresses. A natural compound enriched in apples, ursolic acid reduced muscle atrophy and stimulated muscle hypertrophy in mice. It did so by enhancing skeletal muscle insulin/IGF-I signaling and inhibiting atrophy-associated skeletal muscle mRNA expression. Importantly, ursolic acid's effects on muscle were accompanied by reductions in adiposity, fasting blood glucose, and plasma cholesterol and triglycerides. These findings identify a potential therapy for muscle atrophy and perhaps other metabolic diseases. PMID- 21641546 TI - The circadian clock interacts with metabolic physiology to influence reproductive fitness. AB - Circadian rhythms are regulated by a synchronized system of central and peripheral clocks. Here, we show that a clock in the Drosophila fat body drives rhythmic expression of genes involved in metabolism, detoxification, the immune response, and steroid hormone regulation. Some of these genes cycle even when the fat body clock is disrupted, indicating that they are regulated by exogenous factors. Food is an important stimulus, as limiting food availability to a 6 hr interval each day drives rhythmic expression of genes in the fat body. Restricting food to a time of day when consumption is typically low desynchronizes internal rhythms because it alters the phase of rhythmic gene expression in the fat body without affecting the brain clock. Flies maintained on this paradigm produce fewer eggs than those restricted to food at the normal time. These data suggest that desynchrony of endogenous rhythms, caused by aberrant feeding patterns, affects reproductive fitness. PMID- 21641547 TI - Autophagy regulates cholesterol efflux from macrophage foam cells via lysosomal acid lipase. AB - The lipid droplet (LD) is the major site of cholesterol storage in macrophage foam cells and is a potential therapeutic target for the treatment of atherosclerosis. Cholesterol, stored as cholesteryl esters (CEs), is liberated from this organelle and delivered to cholesterol acceptors. The current paradigm attributes all cytoplasmic CE hydrolysis to the action of neutral CE hydrolases. Here, we demonstrate an important role for lysosomes in LD CE hydrolysis in cholesterol-loaded macrophages, in addition to that mediated by neutral hydrolases. Furthermore, we demonstrate that LDs are delivered to lysosomes via autophagy, where lysosomal acid lipase (LAL) acts to hydrolyze LD CE to generate free cholesterol mainly for ABCA1-dependent efflux; this process is specifically induced upon macrophage cholesterol loading. We conclude that, in macrophage foam cells, lysosomal hydrolysis contributes to the mobilization of LD-associated cholesterol for reverse cholesterol transport. PMID- 21641548 TI - Regulation of yeast chronological life span by TORC1 via adaptive mitochondrial ROS signaling. AB - Here we show that yeast strains with reduced target of rapamycin (TOR) signaling have greater overall mitochondrial electron transport chain activity during growth that is efficiently coupled to ATP production. This metabolic alteration increases mitochondrial membrane potential and reactive oxygen species (ROS) production, which we propose supplies an adaptive signal during growth that extends chronological life span (CLS). In strong support of this concept, uncoupling respiration during growth or increasing expression of mitochondrial manganese superoxide dismutase significantly curtails CLS extension in tor1Delta strains, and treatment of wild-type strains with either rapamycin (to inhibit TORC1) or menadione (to generate mitochondrial ROS) during growth is sufficient to extend CLS. Finally, extension of CLS by reduced TORC1/Sch9p-mitochondrial signaling occurs independently of Rim15p and is not a function of changes in media acidification/composition. Considering the conservation of TOR-pathway effects on life span, mitochondrial ROS signaling may be an important mechanism of longevity regulation in higher organisms. PMID- 21641549 TI - Insulin receptor-related receptor as an extracellular alkali sensor. AB - The insulin receptor-related receptor (IRR), an orphan receptor tyrosine kinase of the insulin receptor family, can be activated by alkaline media both in vitro and in vivo at pH >7.9. The alkali-sensing property of IRR is conserved in frog, mouse, and human. IRR activation is specific, dose-dependent and quickly reversible and demonstrates positive cooperativity. It also triggers receptor conformational changes and elicits intracellular signaling. The pH sensitivity of IRR is primarily defined by its L1F extracellular domains. IRR is predominantly expressed in organs that come in contact with mildly alkaline media. In particular, IRR is expressed in the cell subsets of the kidney that secrete bicarbonate into urine. Disruption of IRR in mice impairs the renal response to alkali loading attested by development of metabolic alkalosis and decreased urinary bicarbonate excretion in response to this challenge. We therefore postulate that IRR is an alkali sensor that functions in the kidney to manage metabolic bicarbonate excess. PMID- 21641550 TI - Mitochondrial phosphatase PTPMT1 is essential for cardiolipin biosynthesis. AB - PTPMT1 was the first protein tyrosine phosphatase found localized to the mitochondria, but its biological function was unknown. Herein, we demonstrate that whole body deletion of Ptpmt1 in mice leads to embryonic lethality, suggesting an indispensable role for PTPMT1 during development. Ptpmt1 deficiency in mouse embryonic fibroblasts compromises mitochondrial respiration and results in abnormal mitochondrial morphology. Lipid analysis of Ptpmt1-deficient fibroblasts reveals an accumulation of phosphatidylglycerophosphate (PGP) along with a concomitant decrease in phosphatidylglycerol. PGP is an essential intermediate in the biosynthetic pathway of cardiolipin, a mitochondrial-specific phospholipid regulating the membrane integrity and activities of the organelle. We further demonstrate that PTPMT1 specifically dephosphorylates PGP in vitro. Loss of PTPMT1 leads to dramatic diminution of cardiolipin, which can be partially reversed by the expression of catalytic active PTPMT1. Our study identifies PTPMT1 as the mammalian PGP phosphatase and points to its role as a regulator of cardiolipin biosynthesis. PMID- 21641551 TI - High-mobility group box 1 is essential for mitochondrial quality control. AB - Mitochondria are organelles centrally important for bioenergetics as well as regulation of apoptotic death in eukaryotic cells. High-mobility group box 1 (HMGB1), an evolutionarily conserved chromatin-associated protein which maintains nuclear homeostasis, is also a critical regulator of mitochondrial function and morphology. We show that heat shock protein beta-1 (HSPB1 or HSP27) is the downstream mediator of this effect. Disruption of the HSPB1 gene in embryonic fibroblasts with wild-type HMGB1 recapitulates the mitochondrial fragmentation, deficits in mitochondrial respiration, and adenosine triphosphate (ATP) synthesis observed with targeted deletion of HMGB1. Forced expression of HSPB1 reverses this phenotype in HMGB1 knockout cells. Mitochondrial effects mediated by HMGB1 regulation of HSPB1 expression serve as a defense against mitochondrial abnormality, enabling clearance and autophagy in the setting of cellular stress. Our findings reveal an essential role for HMGB1 in autophagic surveillance with important effects on mitochondrial quality control. PMID- 21641552 TI - Protein phosphorylation and prevention of cytochrome oxidase inhibition by ATP: coupled mechanisms of energy metabolism regulation. AB - Rapid regulation of oxidative phosphorylation is crucial for mitochondrial adaptation to swift changes in fuels availability and energy demands. An intramitochondrial signaling pathway regulates cytochrome oxidase (COX), the terminal enzyme of the respiratory chain, through reversible phosphorylation. We find that PKA-mediated phosphorylation of a COX subunit dictates mammalian mitochondrial energy fluxes and identify the specific residue (S58) of COX subunit IV-1 (COXIV-1) that is involved in this mechanism of metabolic regulation. Using protein mutagenesis, molecular dynamics simulations, and induced fit docking, we show that mitochondrial energy metabolism regulation by phosphorylation of COXIV-1 is coupled with prevention of COX allosteric inhibition by ATP. This regulatory mechanism is essential for efficient oxidative metabolism and cell survival. We propose that S58 COXIV-1 phosphorylation has evolved as a metabolic switch that allows mammalian mitochondria to rapidly toggle between energy utilization and energy storage. PMID- 21641553 TI - Role for insulin signaling in catecholaminergic neurons in control of energy homeostasis. AB - Dopaminergic midbrain neurons integrate signals on food palatability and food associated reward into the complex control of energy homeostasis. To define the role of insulin receptor (IR) signaling in this circuitry, we inactivated IR signaling in tyrosine hydroxylase (Th)-expressing cells of mice (IR(DeltaTh)). IR inactivation in Th-expressing cells of mice resulted in increased body weight, increased fat mass, and hyperphagia. While insulin acutely stimulated firing frequency in 50% of dopaminergic VTA/SN neurons, this response was abolished in IR(DeltaTh) mice. Moreover, these mice exhibited an altered response to cocaine under food-restricted conditions. Taken together, these data provide in vivo evidence for a critical role of insulin signaling in catecholaminergic neurons to control food intake and energy homeostasis. PMID- 21641554 TI - FGF15/19 regulates hepatic glucose metabolism by inhibiting the CREB-PGC-1alpha pathway. AB - Regulation of hepatic carbohydrate homeostasis is crucial for maintaining energy balance in the face of fluctuating nutrient availability. Here, we show that the hormone fibroblast growth factor 15/19 (FGF15/19), which is released postprandially from the small intestine, inhibits hepatic gluconeogenesis, like insulin. However, unlike insulin, which peaks in serum 15 min after feeding, FGF15/19 expression peaks approximately 45 min later, when bile acid concentrations increase in the small intestine. FGF15/19 blocks the expression of genes involved in gluconeogenesis through a mechanism involving the dephosphorylation and inactivation of the transcription factor cAMP regulatory element-binding protein (CREB). This in turn blunts expression of peroxisome proliferator-activated receptor gamma coactivator-1alpha (PGC-1alpha) and other genes involved in hepatic metabolism. Overexpression of PGC-1alpha blocks the inhibitory effect of FGF15/19 on gluconeogenic gene expression. These results demonstrate that FGF15/19 works subsequent to insulin as a postprandial regulator of hepatic carbohydrate homeostasis. PMID- 21641555 TI - Desnutrin/ATGL is regulated by AMPK and is required for a brown adipose phenotype. AB - While fatty acids (FAs) released by white adipose tissue (WAT) provide energy for other organs, lipolysis is also critical in brown adipose tissue (BAT), generating FAs for oxidation and UCP-1 activation for thermogenesis. Here we show that adipose-specific ablation of desnutrin/ATGL in mice converts BAT to a WAT like tissue. These mice exhibit severely impaired thermogenesis with increased expression of WAT-enriched genes but decreased BAT genes, including UCP-1 with lower PPARalpha binding to its promoter, revealing the requirement of desnutrin catalyzed lipolysis for maintaining a BAT phenotype. We also show that desnutrin is phosphorylated by AMPK at S406, increasing TAG hydrolase activity, and provide evidence for increased lipolysis by AMPK phosphorylation of desnutrin in adipocytes and in vivo. Despite adiposity and impaired BAT function, desnutrin ASKO mice have improved hepatic insulin sensitivity with lower DAG levels. Overall, desnutrin is phosphorylated/activated by AMPK to increase lipolysis and brings FA oxidation and UCP-1 induction for thermogenesis. PMID- 21641556 TI - Thirty years of multiple sequence codes. AB - An overview is presented on the status of studies on multiple codes in genetic sequences. Indirectly, the existence of multiple codes is recognized in the form of several rediscoveries of Second Genetic Code that is different each time. A due credit is given to earlier seminal work related to the codes often neglected in literature. The latest developments in the field of chromatin code are discussed, as well as perspectives of single-base resolution studies of nucleosome positioning, including rotational setting of DNA on the surface of the histone octamers. PMID- 21641557 TI - On the observable transition to living matter. AB - In recent developments in chemistry and genetic engineering, the humble researcher dealing with the origin of life finds her(him)self in a grey area of tackling something that even does not yet have a clear definition agreed upon. A series of chemical steps is described to be considered as the life-nonlife transition, if one adheres to the minimalistic definition: life is self reproduction with variations. The fully artificial RNA system chosen for the exploration corresponds sequence-wise to the reconstructed initial triplet repeats, presumably corresponding to the earliest protein-coding molecules. The demonstrated occurrence of the mismatches (variations) in otherwise complementary syntheses ("self-reproduction"), in this RNA system, opens an experimental and conceptual perspective to explore the origin of life (and its definition), on the apparent edge of the origin. PMID- 21641558 TI - Role of stable isotopes in life--testing isotopic resonance hypothesis. AB - Stable isotopes of most important biological elements, such as C, H, N and O, affect living organisms. In rapidly growing species, deuterium and to a lesser extent other heavy isotopes reduce the growth rate. At least for deuterium it is known that its depletion also negatively impacts the speed of biological processes. As a rule, living organisms "resist" changes in their isotopic environment, preferring natural isotopic abundances. This preference could be due to evolutionary optimization; an additional effect could be due to the presence of the "isotopic resonance". The isotopic resonance phenomenon has been linked to the choice of earliest amino acids, and thus affected the evolution of genetic code. To test the isotopic resonance hypothesis, literature data were analyzed against quantitative and qualitative predictions of the hypothesis. Four studies provided five independent datasets, each in very good quantitative agreement with the predictions. Thus, the isotopic resonance hypothesis is no longer simply plausible; it can now be deemed likely. Additional testing is needed, however, before full acceptance of this hypothesis. PMID- 21641559 TI - On the organizational dynamics of the genetic code. AB - The organization of the canonical genetic code needs to be thoroughly illuminated. Here we reorder the four nucleotides-adenine, thymine, guanine and cytosine-according to their emergence in evolution, and apply the organizational rules to devising an algebraic representation for the canonical genetic code. Under a framework of the devised code, we quantify codon and amino acid usages from a large collection of 917 prokaryotic genome sequences, and associate the usages with its intrinsic structure and classification schemes as well as amino acid physicochemical properties. Our results show that the algebraic representation of the code is structurally equivalent to a content-centric organization of the code and that codon and amino acid usages under different classification schemes were correlated closely with GC content, implying a set of rules governing composition dynamics across a wide variety of prokaryotic genome sequences. These results also indicate that codons and amino acids are not randomly allocated in the code, where the six-fold degenerate codons and their amino acids have important balancing roles for error minimization. Therefore, the content-centric code is of great usefulness in deciphering its hitherto unknown regularities as well as the dynamics of nucleotide, codon, and amino acid compositions. PMID- 21641560 TI - EST-based identification of genes expressed in skeletal muscle of the mandarin fish (Siniperca chuatsi). AB - To enrich the genomic information of the commercially important fish species, we obtained 5,063 high-quality expressed sequence tags (ESTs) from the muscle cDNA database of the mandarin fish (Siniperca chuatsi). Clustering analysis yielded 1,625 unique sequences including 443 contigs (from 3,881 EST sequences) and 1,182 singletons. BLASTX searches showed that 959 unique sequences shared homology to proteins in the NCBI non-redundant database. A total of 740 unique sequences were functionally annotated using Gene Ontology. The 1,625 unique sequences were assigned to Kyoto Encyclopedia of Genes and Genomes reference pathways, and the results indicated that transcripts participating in nucleotide metabolism and amino acid metabolism are relatively abundant in S. chuatsi. Meanwhile, we identified 15 genes to be abundantly expressed in muscle of the mandarin fish. These genes are involved in muscle structural formation and regulation of muscle differentiation and development. The most remarkable gene in S. chuatsi is nuclease diphosphate kinase B, which is represented by 449 EST sequences accounting for 8.86% of the total EST sequences. Our work provides a transcript profile expressed in the white muscle of the mandarin fish, laying down a foundation in better understanding of fish genomics. PMID- 21641561 TI - Computational analysis of drought stress-associated miRNAs and miRNA co regulation network in Physcomitrella patens. AB - miRNAs are non-coding small RNAs that involve diverse biological processes. Until now, little is known about their roles in plant drought resistance. Physcomitrella patens is highly tolerant to drought; however, it is not clear about the basic biology of the traits that contribute P. patens this important character. In this work, we discovered 16 drought stress-associated miRNA (DsAmR) families in P. patens through computational analysis. Due to the possible discrepancy of expression periods and tissue distributions between potential DsAmRs and their targeting genes, and the existence of false positive results in computational identification, the prediction results should be examined with further experimental validation. We also constructed an miRNA co-regulation network, and identified two network hubs, miR902a-5p and miR414, which may play important roles in regulating drought-resistance traits. We distributed our results through an online database named ppt-miRBase, which can be accessed at http://bioinfor.cnu.edu.cn/ppt_miRBase/index.php. Our methods in finding DsAmR and miRNA co-regulation network showed a new direction for identifying miRNA functions. PMID- 21641562 TI - Identification of protein-coding regions in DNA sequences using a time-frequency filtering approach. AB - Accurate identification of protein-coding regions (exons) in DNA sequences has been a challenging task in bioinformatics. Particularly the coding regions have a 3-base periodicity, which forms the basis of all exon identification methods. Many signal processing tools and techniques have been applied successfully for the identification task but still improvement in this direction is needed. In this paper, we have introduced a new promising model-independent time-frequency filtering technique based on S-transform for accurate identification of the coding regions. The S-transform is a powerful linear time-frequency representation useful for filtering in time-frequency domain. The potential of the proposed technique has been assessed through simulation study and the results obtained have been compared with the existing methods using standard datasets. The comparative study demonstrates that the proposed method outperforms its counterparts in identifying the coding regions. PMID- 21641563 TI - TAAPP: Tiling Array Analysis Pipeline for Prokaryotes. AB - High-density tiling arrays provide closer view of transcription than regular microarrays and can also be used for annotating functional elements in genomes. The identified transcripts usually have a complex overlapping architecture when compared to the existing genome annotation. Therefore, there is a need for customized tiling array data analysis tools. Since most of the initial tiling arrays were conducted in eukaryotes, data analysis methods are well suited for eukaryotic genomes. For using whole-genome tiling arrays to identify previously unknown transcriptional elements like small RNA and antisense RNA in prokaryotes, existing data analysis tools need to be tailored for prokaryotic genome architecture. Furthermore, automation of such custom data analysis workflow is necessary for biologists to apply this powerful platform for knowledge discovery. Here we describe TAAPP, a web-based package that consists of two modules for prokaryotic tiling array data analysis. The transcript generation module works on normalized data to generate transcriptionally active regions (TARs). The feature extraction and annotation module then maps TARs to existing genome annotation. This module further categorizes the transcription profile into potential novel non-coding RNA, antisense RNA, gene expression and operon structures. The implemented workflow is microarray platform independent and is presented as a web based service. The web interface is freely available for academic use at http://lims.lsbi.mafes.msstate.edu/TAAPP-HTML/. PMID- 21641564 TI - Impact of instruction on the acquisition of sequence knowledge in a sensorimotor task. AB - We examined whether and to what extent a sequence of finger movements can be learned and transferred to the untrained hand according to the muscle homology depending on the relative salience of response locations and effectors. Participants performed a discrete sequence production task, in which they were asked to learn a sequence of either key locations or of finger movements. Each training block was followed by a transfer block in which responding with the opposite hand was required. Before the last transfer block participants received an unexpected instruction. They had to reproduce the sequence of key locations instead of the sequence of finger movements and conversely, the sequence of finger movements instead of the sequence of key locations. The results do not support the existence of a sequence representation for the order of finger movements irrespective of the hand used. PMID- 21641565 TI - [Acute generalized exanthematous pustulosis due to milk thistle (Silybum marianum) tea]. PMID- 21641566 TI - [Pigmentation of the fungiform papillae of the tongue: a report of 2 cases]. PMID- 21641567 TI - [History of Actas Dermo-Sifiliograficas, part I: 1909-1959]. AB - Actas Dermo-Sifiliograficas was born in May 1909. At first, issues appeared in step with the academic year, but publication began to follow the calendar year in 1957. Volume 18 was skipped in 1926-7 in an effort to correct confusion in the numbering of volumes and pages of earlier issues. October 1928 saw the journal grow from 6 issues per year to 9. Although the Spanish Civil War brought publication to a halt during the 1936-7 academic year, Actas Dermo Sifiliograficas was one of the first Spanish scientific journals to recover from the conflict. The initial print run of 100 copies was increased to 700 after the war. The content evolved over time: while originally conceived to provide a strict account of sessions of the Spanish Academy of Dermatology and Venereology (AEDV) -originally known as the Spanish Academy of Dermatology and Syphilology- the journal gradually came to include review articles, case reports, a section summarizing the content of international journals, news and various other types of writing. The editorial board and the association's board of directors were one and the same for many years. According to the earliest charter, the editor-in chief was also the president of the association and the associate editor was the association's vice-president. The subjects of articles provide a faithful portrait of how the specialty has changed. Syphilis, a main concern before the introduction of penicillin in the 1940s, was sidelined afterwards. The appearance of 20th-century pharmaceuticals such as salvarsan, sulfa drugs, thiazides, and corticosteroids were soon reflected in the number of articles describing their use. Certain original contributions by Spanish authors to international dermatology first appeared in Actas Dermo-Sifiliograficas. Examples are Azua's description of pseudoepithelioma and Covisa and Bejarano's of chancriform pyoderma. Volume 50 (1959), which included accounts of the 50th anniversary of the association and the journal, closed with a biography of Enrique Alvarez Sainz de Aja, the only founding member still living at that time. PMID- 21641568 TI - [Esomeprazole-induced subacute cutaneous lupus erythematosus]. PMID- 21641569 TI - Cardiopulmonary limited ultrasound examination for "quick-look" bedside application. AB - Although taking a "quick look" at the heart using a small ultrasound device is now feasible, a formal ultrasound imaging protocol to augment the bedside physical examination has not been developed. Therefore, we sought to evaluate the diagnostic accuracy and prognostic value of a cardiopulmonary limited ultrasound examination (CLUE) using 4 simplified diagnostic criteria that would screen for left ventricular dysfunction (LV), left atrial (LA) enlargement, inferior vena cava plethora (IVC+), and ultrasound lung comet-tail artifacts (ULC+) in patients referred for echocardiography. The CLUE was tested by interpretation of only the parasternal LV long-axis, subcostal IVC, and 2 lung apical views in each of 1,016 consecutive echocardiograms performed with apical lung imaging. For inpatients, univariate and multivariate logistic regression analyses were performed to assess the relations between mortality, CLUE findings, age, and gender. In this echocardiographic referral series, 78% (n = 792) were inpatient and 22% (n = 224) were outpatient. The CLUE criteria demonstrated a sensitivity, specificity, and accuracy for a LV ejection fraction of <=40% of 69%, 91%, and 89% and for LA enlargement of 75%, 72%, and 73%, respectively. CLUE findings of LV dysfunction, LA enlargement, IVC+, and ULC+ were seen in 16%, 53%, 34%, and 28% of inpatients. The best multivariate logistic model contained 3 predictors of in-hospital mortality: ULC+, IVC+ and male gender, with adjusted odds ratios (95% confidence intervals) of 3.5 (1.4 to 8.8), 5.8 (2.1 to 16.4), and 2.3 (0.9 to 5.8), respectively. In conclusion, a CLUE consisting of 4 quick-look "signs" has reasonable diagnostic accuracy for bedside use and contains prognostic information. PMID- 21641570 TI - Uterine artery embolization versus surgery in the treatment of symptomatic fibroids: a systematic review and metaanalysis. AB - OBJECTIVE: To summarize the evidence on short-, mid-, and long-term results up to 5 years of uterine artery embolization in comparison to surgery. STUDY DESIGN: We searched the CENTRAL, MEDLINE and EMBASE databases for randomized clinical trials comparing uterine artery embolization with hysterectomy/myomectomy in premenopausal women with heavy menstrual bleeding caused by symptomatic uterine fibroids, written from September 1995 to November 2010. Two reviewers independently assessed methodologic quality and extracted data from included trials. RESULTS: Four randomized controlled trials with a total of 515 patients were included. On the short-term, uterine artery embolization showed fewer blood loss, shorter hospital stay, and quicker resumption of work. Mid- and long-term results showed comparable health-related quality of life results and a higher reintervention rate in the uterine artery embolization group, whereas both groups were equally satisfied. CONCLUSION: Uterine artery embolization has short-term advantages over surgery. On the mid- and long-term the benefits were similar, except for a higher reintervention rate after uterine artery embolization. PMID- 21641571 TI - Long-term neurodevelopmental outcome in TTTS in the Eurofoetus trial. PMID- 21641572 TI - A methyl-deviator epigenotype of estrogen receptor-positive breast carcinoma is associated with malignant biology. AB - We broadly profiled DNA methylation in breast cancers (n = 351) and benign parenchyma (n = 47) for correspondence with disease phenotype, using FFPE diagnostic surgical pathology specimens. Exploratory analysis revealed a distinctive primary invasive carcinoma subclass featuring extreme global methylation deviation. Subsequently, we tested the correlation between methylation remodeling pervasiveness and malignant biological features. A methyl deviation index (MDI) was calculated for each lesion relative to terminal ductal lobular unit baseline, and group comparisons revealed that high-grade and short survival estrogen receptor-positive (ER(+)) cancers manifest a significantly higher MDI than low-grade and long-survival ER(+) cancers. In contrast, ER(-) cancers display a significantly lower MDI, revealing a striking epigenomic distinction between cancer hormone receptor subtypes. Kaplan-Meier survival curves of MDI-based risk classes showed significant divergence between low- and high-risk groups. MDI showed superior prognostic performance to crude methylation levels, and MDI retained prognostic significance (P < 0.01) in Cox multivariate analysis, including clinical stage and pathological grade. Most MDI targets individually are significant markers of ER(+) cancer survival. Lymphoid and mesenchymal indexes were not substantially different between ER(+) and ER(-) groups and do not explain MDI dichotomy. However, the mesenchymal index was associated with ER(+) cancer survival, and a high lymphoid index was associated with medullary carcinoma. Finally, a comparison between metastases and primary tumors suggests methylation patterns are established early and maintained through disease progression for both ER(+) and ER(-) tumors. PMID- 21641573 TI - Beyond consent--improving understanding in surgical patients. AB - BACKGROUND: Little is known of the actual understanding that underlies patient choices with regard to their surgical treatment. This review explores current knowledge of patient understanding and techniques that may be used to improve this understanding. METHODS: MEDLINE and PubMed were searched using the terms "patient understanding," "patient comprehension," "consent," "video," "multimedia," "patient information leaflet," "internet," "test-feedback," "extended discussion," "shared decision making," and "decision aid." All retrieved peer-reviewed studies were included in the review. RESULTS: Understanding in surgical patients is poor. There is little evidence to support the use of information leaflets, although multimedia appears to be effective in improving patient understanding. The internet is not used effectively as an aid to consent by health care providers. Patients with lower educational levels may gain most from additional interventions. Improving patient understanding does not impact on their satisfaction with the treatment they have received but may reduce periprocedural anxiety. CONCLUSIONS: There is a need for greater awareness of patients' information needs, and novel approaches that may enhance decision making through improved understanding are required. PMID- 21641574 TI - Does plasma IGF-BP3 measurement contribute to the diagnosis of growth hormone deficiency in children? AB - OBJECTIVE: To audit the contribution of plasma IGF-PB3 measurement to the diagnosis of growth hormone deficiency (GHD) in children. POPULATION AND METHODS: Retrospective case study including boys and girls aged 0 to 18 years who attended our paediatric endocrinology clinic for short stature and/or post-irradiation follow-up, and had at least one GH provocative testing. Children with hypothyroidism, Laron or Kowarski syndromes, severe malnutrition, chronic renal failure and liver failure were excluded. RESULTS: Fifty-eight children were enrolled and grouped as GHD [+] (19 cases) and GDH [-] (39 cases). IGF-I and IGF BP3 assay was carried out in 88% and 62% cases respectively, both groups were comparable for age, sex, BMI, target height, pubertal stage and bone age. There was a significant difference in peak GH between GDH [-] and GHD [+] groups (41.8 mUI/L +/- 21.7 versus 11.5 +/- 5.9 mUI/L, P<0.00001, respectively). No difference was found between groups with regards to IGF-I Z-scores and IGF-BP3 Z-scores. There was, however, a positive correlation between IGF-I Z-scores and IGF-BP3 Z scores (r=0.50; P<0.0016). IGF-BP3 measurement could not differentiate between GHD [+] and GHD [-] groups. CONCLUSIONS: Measurement of plasma IGF-BP3 level contributes poorly to the diagnosis of GHD. We do not recommend it in routine use. PMID- 21641575 TI - Non-ultrasound-guided ethanol sclerotherapy for the treatment of thyroid cysts. AB - OBJECTIVE: Ethanol injection under ultrasound (US) guidance has been proposed as an alternative to surgery in the treatment of recurrent benign thyroid cysts following aspiration. We aimed to set up a new procedure of ethanol sclerotherapy without US guidance for the treatment of pure thyroid cysts in order to make this useful treatment, available to more patients, more particularly when access to centers specialized in thyroid ultrasonography is limited. PATIENTS AND METHODS: Nine patients with recurrent large thyroid cysts following aspiration, and showing symptoms of compression and/or cosmetic complaints were treated by ethanol injection without US guidance and followed for up to 11 years. RESULTS: After ethanol injection, mean cyst volume was significantly reduced (9.9 +/- 13.6 vs. 31.3 +/- 34.1 ml, P=0.007) and the mean percentage volume reduction was 72.7%. A size reduction of the thyroid lesion more than 50% was achieved in eight of the nine patients (89%). Compressive symptoms and cosmetic complaints totally disappeared after sclerotherapy in all patients. During a mean follow-up of 48 months (ranging from 12 to 135 months), no recurrences were observed. The treatment was well tolerated with no major side effects. CONCLUSION: Non-US guided ethanol sclerotherapy is a safe and "easy-to-use" procedure to treat benign thyroid cysts effectively. Because this new treatment does not need US guidance, it can be performed by endocrinologists during outpatient visits. This new procedure may be useful in some areas, such as developing countries, where access to US examination is limited. PMID- 21641577 TI - Good practice guide for cervical ultrasound scan and echo-guided techniques in treating differentiated thyroid cancer of vesicular origin. AB - Good practice guide for cervical ultrasound scan and echo-guided techniques in treating differentiated thyroid cancer of vesicular origin. American, European and French Recommendations for the treatment of differentiated vesicular thyroid cancer were recently published. Cervical ultrasound scanning is now considered a key examination in the follow-up of these cancers. This examination is noninvasive, easy to perform and to obtain, is not costly, but remains operator dependent. To date, there are no recommendations published that assemble all the technical aspects, results, indications and the limits of this examination in the initial medical report and the follow-up of these cancers. In order to standardise the procedure and validate the quality of the examination, a workgroup made up of a panel of experts particularly involved in carrying out ultrasound scans was set up. The aim was to draw up a good practice guide for performing cervical ultrasound scans and echo-guided techniques in treating patients with differentiated thyroid cancer of vesicular origin. The main objectives are to: (a) standardise the procedure and reports, (b) define the criteria for establishing whether lesions identified during a cervical ultrasound scan are malignant or benign, (c) standardise the indications for carrying out cytological tests and an in situ assay of markers, (d) help doctors to select the patients who ought to receive a cervical ultrasound scan and or cytological tests, (e) discuss how frequently the examinations should be carried out depending on the risk of recurrence. PMID- 21641578 TI - Odontoblast-like cell differentiation and dentin formation induced with TGF beta1. AB - OBJECTIVE: To investigate the inductive potential of scaffold material combing with transforming growth factor-beta1 (TGF-beta1), and to induce odontoblast differentiation and dentin formation from dental pulp cells both in vitro and in vivo. METHODS: Primarily cultured dental pulp cells were used for MTT, ALP activity assay and Alizarin red staining in the presence of TGF-beta1. Pelleted cells were put on the filters combining with or not with TGF-beta1 and cultured in vitro or in vivo. The in vitro and in vivo cell response and tissue formation were analysed with Haematoxylin-Eosin (HE), transmission electron microscopy (TEM) and immunohistochemical staining. RESULTS: TGF-beta1 increased the mineralization and ALP activity of dental pulp cells as revealed by Alizarin red staining and ALP activity assay. After in vitro culture for 7 days, cells polarized in the TGF-beta1 group and expressed dentin sialoprotein (DSP), osteopontin (OPN) and type I collagen (Col I). After in vivo transplantation for 7 days, columnar odontoblast formed on the surface of filter in experimental group, and tubular dentin expressing DSP formed after 3 months transplantation. CONCLUSION: It was concluded that TGF-beta1 combining with transfilter could induce odontoblast differentiation and dentin formation. Our results implied that suitable substrate for the progenitors of odontoblast to anchor on and inductive signals to initiate the differentiation of odontoblast should be taken into consideration when designing scaffold material for inducing dentin tissue engineering. PMID- 21641579 TI - Pressure induces interleukin-6 expression via the P2Y6 receptor in human dental pulp cells. AB - BACKGROUND AND OBJECTIVE: An increase in intrapulpal pressure occurs during inflammation and restorative procedures; however, the role of the pressure on human dental pulp cell (HDPC) is not yet clarified. In this study, the effect of pressure on interleukin-6 (IL-6) expression of HDPCs was examined. DESIGN: HDPCs were applied with pressure (0.7-1.4 g/cm(2)). The level of IL-6 mRNA and protein release was determined by RT-PCR and ELISA, respectively. The signalling pathways were investigated using inhibitors, antagonists, and small interfering RNA. RESULTS: The results showed that pressure up-regulated IL-6 mRNA expression and protein release in a time- and dose-dependent manner. The implication of P2Y receptor was exhibited by a significant inhibition of pressure-induced IL-6 expression by suramin, an antagonist for the non-specific purinergic receptor family. Using loss of function experiments, we showed MRS2578 (a specific P2Y6 antagonist), as well as P2Y6 small interfering RNA, abolished pressure-induced IL 6, whilst MRS2179 (a specific P2Y1 antagonist) and NF449 (a P2X1, P2X3, P2Y1, and P2Y2 antagonist) had no effect. Finally, we demonstrated that either the conditioned medium collected from pressurised dental pulp cells or addition of UDP, a selective agonist of P2Y6, up-regulated IL-6 expression in HDPCs. CONCLUSIONS: These results indicate that pressure could induce IL-6 expression through the P2Y6 receptor in HDPCs, leading to a new insight of the role of pressure on cytokine release during pulpal inflammatory process. PMID- 21641580 TI - Randomised controlled trial of a guided self-help treatment on the Internet for binge eating disorder. AB - Binge eating disorder (BED) is a common and under-treated condition with major health implications. Cognitive behavioural therapy (CBT) self-help manuals have proved to be efficient in BED treatment. Increasing evidence also support the use of new technology to improve treatment access and dissemination. This is the first randomised controlled study to evaluate the efficacy of an Internet guided self-help treatment programme, based on CBT, for adults with threshold and subthreshold BED. Seventy-four women were randomised into two groups. The first group received the six-month online programme with a six-month follow-up. The second group was placed in a six-month waiting list before participating in the six-month intervention. Guidance consisted of a regular e-mail contact with a coach during the whole intervention. Binge eating behaviour, drive for thinness, body dissatisfaction and interoceptive awareness significantly improved after the Internet self-help treatment intervention. The number of objective binge episodes, overall eating disorder symptoms score and perceived hunger also decreased. Improvements were maintained at six-month follow-up. Dropouts exhibited more shape concern and a higher drive for thinness. Overall, a transfer of CBT-based self-help techniques to the Internet was well accepted by patients, and showed positive results for eating disorders psychopathology. PMID- 21641582 TI - A study of the bidirectional association between hippocampal volume on magnetic resonance imaging and depression in the elderly. AB - BACKGROUND: Hippocampal volume loss on magnetic resonance imaging (MRI) has been reported in patients with depression. It is uncertain whether a small hippocampus renders a person vulnerable to develop depression or whether it is a consequence of depression. In this study, we addressed whether smaller baseline MRI hippocampal volumes increase the risk of incident depression. We also examined whether depressive symptoms at baseline were associated with decline in hippocampal volume during follow-up. METHODS: Data were obtained in a prospective population-based study over a 10-year period. A sample of 514 nondemented persons aged 60 to 90 years underwent baseline measurements in 1995-1996 including three dimensional MRI scans for assessment of hippocampal volumes and depressive symptoms (measured with Center for Epidemiologic Studies Depression Scale). Follow-up MRIs were made in 1999-2000 and in 2006. Incident depression was identified through standardized psychiatric examinations and continuous monitoring of medical and pharmaceutical records. RESULTS: During a mean follow up of 6.8 years per person (range .07-10.01 years), 135 of the 514 persons developed a clinically relevant episode of incident depressive symptoms. There was no association between baseline hippocampal volumes and incident depression (hazard ratio per SD decrease of average hippocampal volume .98 [.81-1.19], p = .84). A baseline Center for Epidemiologic Studies Depression Scale score of 16 or higher predicted a faster rate of decline in hippocampal volume. Also, incident depression was accompanied by a faster decline in left hippocampal volume. CONCLUSIONS: This study provides no evidence that a small hippocampal volume precedes the development of late-life depression. Depression, however, may lead to a faster rate of hippocampal volume decline. PMID- 21641581 TI - Meta-analysis of cytokine alterations in schizophrenia: clinical status and antipsychotic effects. AB - BACKGROUND: Schizophrenia is associated with immune system dysfunction, including aberrant cytokine levels. We performed a meta-analysis of these associations, considering effects of clinical status and antipsychotic treatment following an acute illness exacerbation. METHODS: We identified articles by searching PubMed, PsychInfo, and Institute for Scientific Information and the reference lists of identified studies. RESULTS: Forty studies met the inclusion criteria. Effect sizes were similar for studies of acutely relapsed inpatients (AR) and first episode psychosis (FEP). Interleukin (IL)-1beta, IL-6, and transforming growth factor-beta (TGF-beta) appeared to be state markers, as they were increased in AR and FEP (p < .001 for each) and normalized with antipsychotic treatment (p < .001, p = .008, and p = .005, respectively). In contrast, IL-12, interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), and soluble IL-2 receptor (sIL-2R) appeared to be trait markers, as levels remained elevated in acute exacerbations and following antipsychotic treatment. There was no difference in IL-6 levels between stable medicated outpatients and control subjects (p = .69). In the cerebrospinal fluid, IL-1beta was significantly decreased in schizophrenia versus controls (p = .01). CONCLUSIONS: Similar effect sizes in AR and FEP suggest that the association between cytokine abnormalities and acute exacerbations of schizophrenia is independent of antipsychotic medications. While some cytokines (IL-1beta, IL-6, and TGF-beta) may be state markers for acute exacerbations, others (IL-12, IFN-gamma, TNF-alpha, and sIL-2R) may be trait markers. Although these results could provide the basis for future hypothesis testing, most studies did not control for potential confounding factors such as body mass index and smoking. PMID- 21641583 TI - Mega cisterna magna associated with recurrent catatonia: a case report. PMID- 21641585 TI - Synthesis of the conjugation ready, downstream disaccharide fragment of the O-PS of Vibrio cholerae O:139. AB - The linker-equipped disaccharide, 8-amino-3,6-dioxaoctyl 2,6-dideoxy-2-acetamido 3-O-beta-D-galactopyranosyluronate-beta-D-glucopyranoside (10), was synthesized in eight steps from acetobromogalactose and ethyl 4,6-O-benzylidene-2-deoxy-2 trichloroacetamido-1-thio-beta-D-glucopyranoside. The hydroxyl group present at C 4(II) in the last intermediate, 8-azido-3,6-dioxaoctyl 4-O-benzyl-6-bromo-2,6 dideoxy-2-trichloroacetamido-3-O-(benzyl 2,3-di-O-benzyl-beta-D galactopyranosyluronate)-beta-D-glucopyranoside (9), is positioned to allow further build-up of the molecule and, eventually, construction of the complete hexasaccharide. Global deprotection (9->10) was done in one step by catalytic hydrogenolysis over palladium-on-charcoal. PMID- 21641584 TI - Breaking down the barrier: the effects of HIV-1 on the blood-brain barrier. AB - Human immunodeficiency virus type 1 (HIV-1) primarily infects CD4(+) T cells and cells of the monocyte-macrophage lineage, resulting in immunodeficiency in an infected patient. Along with this immune deficiency, HIV-1 has been linked to a number of neurological symptoms in the absence of opportunistic infections or other co-morbidities, suggesting that HIV-1 is able to cross the blood-brain barrier (BBB), enter the central nervous system (CNS), and cause neurocognitive impairment. HIV-1-infected monocyte-macrophages traverse the BBB and enter the CNS throughout the course of HIV-1 disease. Once in the brain, both free virus and virus-infected cells are able to infect neighboring resident microglia and astrocytes and possibly other cell types. HIV-1-infected cells in both the periphery and the CNS give rise to elevated levels of viral proteins, including gp120, Tat, and Nef, and of host inflammatory mediators such as cytokines and chemokines. It has been shown that the viral proteins may act alone or in concert with host cytokines and chemokines, affecting the integrity of the BBB. The pathological end point of these interactions may facilitate a positive feedback loop resulting in increased penetration of HIV into the CNS. It is proposed in this review that the dysregulation of the BBB during and after neuroinvasion is a critical component of the neuropathogenic process and that dysregulation of this protective barrier is caused by a combination of viral and host factors including secreted viral proteins, components of the inflammatory process, the aging process, therapeutics, and drug or alcohol abuse. PMID- 21641586 TI - Enzymatic synthesis of dimeric glycomimetic ligands of NK cell activation receptors. AB - This work reveals new structural relationships in the complex process of the interaction between activation receptors of natural killer cells (rat NKR-P1, human CD69) and novel bivalent carbohydrate glycomimetics. The length, glycosylation pattern and linker structure of receptor ligands were examined with respect to their ability to precipitate the receptor protein from solution, which simulates the in vivo process of receptor aggregation during NK cell activation. It was found that di-LacdiNAc triazole compounds show optimal performance, reaching up to 100% precipitation of the present protein receptors, and achieving high immunostimulatory activities without any tendency to trigger activation induced apoptosis. In the synthesis of the compounds tested, two enzymatic approaches were applied. Whereas a beta-N-acetylhexosaminidase could only glycosylate one of the two acceptor sites available with yields below 10%, the Y284L mutant of human placental beta1,4-galactosyltransferase-1 worked as a perfect synthetic tool, accomplishing even quantitative glycosylation at both acceptor sites and with absolute regioselectivity for the C-4 position. This work insinuates new directions for further ligand structure optimisation and demonstrates the strong synthetic potential of the mutant human placental beta1,4 galactosyltransferase-1 in the synthesis of multivalent glycomimetics and glycomaterials. PMID- 21641587 TI - Synthesis of glycolipopeptidic building blocks for carbohydrate receptor discovery. AB - A class of glycolipopeptides for use as building blocks for a new type of dynamic combinatorial library is reported. The members of the library consist of a variable carbohydrate moiety, coded amino acids, and lipoamino acids in order to convert them into amphiphiles. Glycolipopeptidic amphiphiles interact through non covalent bonding when mixed together in aqueous phase and form micelles in dynamic close-packed fluid mosaic arrays. The head groups of amphiphiles are exposed on the micelle surface, providing entities which could be screened in biological assays to find the most potent combination of building blocks in order to identify new bioactive carbohydrate ligands. PMID- 21641588 TI - Cancer-testis antigen, BORIS based vaccine delivered by dendritic cells is extremely effective against a very aggressive and highly metastatic mouse mammary carcinoma. AB - Here, we analyze for the first time the immunological and therapeutic efficacy of a dendritic cell (DC) vaccine based on a cancer-testis antigen, Brother of regulator of imprinted sites (BORIS), an epigenetically acting tumor-promoting transcription factor. Vaccination of mice with DC loaded with truncated form of BORIS (DC/mBORIS) after 4T1 mammary tumor implantation induced strong anti-cancer immunity, inhibited tumor growth (18.75% of mice remained tumor-free), and dramatically lowered the number of spontaneous clonogenic metastases (50% of mice remained metastases-free). Higher numbers of immune effector CD4 and CD8 T cells infiltrated the tumors of vaccinated mice vs. control animals. Vaccination significantly decreased the number of myeloid-derived suppressor cells (MDSCs) infiltrating the tumor sites, but not MDSCs in the spleens of vaccinated animals. These data suggest that DC-based mBORIS vaccination strategies have significant anti-tumor activity in a therapeutic setting and will be more effective when combined with agents to attenuate tumor-associated immune suppression. PMID- 21641589 TI - Temperaments mediate suicide risk and psychopathology among patients with bipolar disorders. AB - BACKGROUND: Several studies have demonstrated that bipolar II (BD-II) disorder represents a quite common, distinct form of major mood disorders that should be separated from bipolar I (BD-I) disorder. The aims of this cross-sectional study were to assess temperament and clinical differences between patients with BD-I and BD-II disorders and to assess whether temperament traits are good predictors of hopelessness in patients with bipolar disorder, a variable highly associated with suicidal behavior and ideation. METHOD: Participants were 216 consecutive inpatients (97 men and 119 women) with a Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revision (DSM-IV-TR), BD who were admitted to the Sant'Andrea Hospital's psychiatric ward in Rome (Italy). Patients completed the Temperament Evaluation of Memphis, Pisa, Paris, and San Diego- Autoquestionnaire, the Beck Hopelessness Scale (BHS), the Mini International Neuropsychiatric Interview (MINI), and the Gotland Scale of Male Depression. RESULTS: Patients with BD-II had higher scores on the BHS (9.78 +/- 5.37 vs 6.87 +/- 4.69; t(143.59) = -3.94; P < .001) than patients with BD-I. Hopelessness was associated with the individual pattern of temperament traits (ie, the relative balance of hyperthymic vs cyclothymic-irritable-anxious-dysthmic). Furthermore, patients with higher hopelessness (compared with those with lower levels of hopelessness) reported more frequently moderate to severe depression (87.1% vs 38.9%; P < .001) and higher MINI suicidal risk. CONCLUSION: Temperaments are important predictors both of suicide risk and psychopathology and may be used in clinical practice for better delivery of appropriate care to patients with bipolar disorders. PMID- 21641590 TI - Reconstruction of patient-specific femurs using X-ray and sparse CT images. AB - Three-dimensional patient-specific bone models have been used in computer-aided planning and biomechanical analysis of orthopaedic surgeries. Reconstruction methods using X-ray images have been developed recently. However, these reconstruction methods have limited ability to generate femur models with severe rotational deformities. In this study, a new X-ray-based reconstruction method was proposed using the free form deformation method with two X-ray images and three CT images. The obtained femur model is closer to a CT-based 3D femur model in comparison with the reconstruction method using only X-ray images. This method will have benefits for many clinical and biomechanical applications. PMID- 21641591 TI - Damages at Japanese assisted reproductive technology clinics by the Great Eastern Japan Earthquake of 2011. AB - The Great Eastern Japan Earthquake of 2011 and consequent tsunami affected many IVF clinics in northeastern Japan. Twelve clinics lost at least one embryo, but all the frozen embryos stored in liquid nitrogen tanks were safe. Emergency power supply is the first priority issue in an unpredictable natural disaster. PMID- 21641592 TI - Effect of cyclosporine pretreatment on mitochondrial function in vitrified bovine mature oocytes. AB - Vitrification had a significantly negative impact on the mitochondrial function of bovine oocytes. However, 40 MUg/mL cyclosporine pretreatment before vitrification contributed greatly to maintaining mitochondrial membrane potential and adenosine triphosphate content, decreasing reactive oxygen species level, and thereby increasing the developmental ability of vitrified oocytes. PMID- 21641593 TI - The D-chiro-inositol paradox in the ovary. AB - The D-chiro-inositol-to-myo-inositol ratio is regulated by an insulin-dependent epimerase. Enzyme activity varies among tissues, likely owing to the specific needs of the two different molecules. We hypothesize that in the ovaries of polycystic ovary syndrome patients, epimerase activity is enhanced, leading to a local myo-inositol deficiency which in turn is responsible for the poor oocyte quality. PMID- 21641594 TI - Steroid hormone hypersensitivity: clinical presentation and management. AB - Hypersensitivity to estrogens and progestogens is often undiagnosed. The condition has many manifestations, including premenstrual syndrome, dysmenorrhea, and impaired fertility. Diagnosis is confirmed by skin testing for inflammatory responses to small doses of the hormone, and desensitization with small doses of the hormone is the most appropriate form of management. PMID- 21641595 TI - Age does not influence the effect of embryo fragmentation on successful blastocyst development. AB - We evaluated the rate of blastocyst development in day 3 embryos with appropriate cellular division and investigated whether maternal age modified the effect of embryo fragmentation on blastulation. Our data showed a significant negative correlation between the degree of embryo fragmentation and rate of blastocyst development, but age did not exert an effect on the degree of fragmentation in embryos with appropriate cleaving status, nor did it modify the significant effect embryo fragmentation had on blastocyst formation. PMID- 21641596 TI - Late sequelae of retained foreign bodies after world war II missile injuries. AB - A number of people injured during the second world war harbour foreign bodies such as grenade splinters or bullets in some part of the body. Most of these metal fragments remain clinically silent. Some of them, however, may cause delayed complications. The purpose of this study was to determine the characteristics of delayed complications associated with foreign bodies after world war II injuries. 159 patients with retained foreign bodies after world war II injuries were retrospectively identified radiologically in our data bases in the time interval from 1997 to 2009. Diverse delayed complications secondary to the metal objects were diagnosed in 3 cases (2%): one patient with grenade splinter migration into the choledochal duct, one case with pseudotumoural tissue reaction, and one patient with late osteomyelitis. The time from injury to clinical presentation varied from 56 to 61 years. PubMed and Medline were screened for additional cases with delayed sequelae after foreign body acquisition during the 2nd world war. A 30 year search period from 1980 up to date was selected. 15 cases were identified here. Our study demonstrates that health consequences of the 2nd world war extend into the present time, and therefore physicians should be aware of the presence of hidden foreign bodies and their different possible late reactions. PMID- 21641597 TI - Proximal tibia fractures and intramedullary nailing: The impact of nail trajectory to varus/valgus deformity. PMID- 21641599 TI - Solution cathode glow discharge induced vapor generation of mercury and its application to mercury speciation by high performance liquid chromatography atomic fluorescence spectrometry. AB - A novel solution cathode glow discharge (SCGD) induced vapor generation was developed as interface to on-line couple high-performance liquid chromatography (HPLC) with atomic fluorescence spectrometry (AFS) for the speciation of inorganic mercury (Hg(2+)), methyl-mercury (MeHg) and ethyl-mercury (EtHg). The decomposition of organic mercury species and the reduction of Hg(2+) could be completed in one step with this proposed SCGD induced vapor generation system. The vapor generation is extremely rapid and therefore is easy to couple with flow injection (FI) and HPLC. Compared with the conventional HPLC-CV-AFS hyphenated systems, the proposed HPLC-SCGD-AFS system is very simple in operation and eliminates auxiliary redox reagents. Parameters influencing mercury determination were optimized, such as concentration of formic acid, discharge current and argon flow rate. The method detection limits for HPLC-SCGD-AFS system were 0.67 MUg L( 1) for Hg(2+), 0.55 MUg L(-1) for MeHg and 1.19 MUg L(-1) for EtHg, respectively. The developed method was validated by determination of certified reference material (GBW 10029, tuna fish) and was further applied for the determination of mercury in biological samples. PMID- 21641598 TI - Serum apolipoprotein B-48 levels are correlated with carotid intima-media thickness in subjects with normal serum triglyceride levels. AB - BACKGROUND: Postprandial hyperlipidemia (PPHL) is an independent risk factor for coronary heart disease (CHD) which is based on the accumulation of chylomicrons (CM) and CM remnants containing apolipoprotein B-48 (apoB-48). Since atherosclerotic cardiovascular diseases are frequently observed even in subjects with normal serum triglyceride (TG) level, the correlation between fasting apoB 48 containing lipoproteins and carotid intima-media thickness (IMT) was analyzed in subjects with normal TG levels. METHODS: From subjects who took their annual health check at the Osaka Police Hospital (n=245, male), one-hundred and sixty four male subjects were selected to take part in this study; the excluding factors were: systolic blood pressure >= 140 mmHg, intake of antihypertensive or antihyperlipidemic drugs, or age >65 years. The association between biochemical markers and IMT was analyzed and independent predictors of max-IMT were determined by multiple regression analysis in all subjects and in groups N-1 (TG<100mg/dl, n=58), N-2 (100 <= TG<150 mg/dl, n=53) and H (150 <= TG mg/dl, n=53), respectively. RESULTS: Fasting total cholesterol, LDL-cholesterol, HDL cholesterol, apoB-100 and lnRemL-C (remnant lipoprotein-cholesterol) levels were not correlated with max-IMT, but lnTG and lnapoB-48 were significantly correlated with max-IMT in all subjects. LnapoB-48 and apoB-48/TG ratio were significantly correlated with max-IMT in group N-2. By multiple regression analysis, age and lnapoB-48 were independent variables associated with max-IMT in group N-2. CONCLUSION: Serum apoB-48 level might be a good marker for the detection of early atherosclerosis in middle-aged subjects with normal-range levels of blood pressure and TG. PMID- 21641600 TI - Improved enantiomer resolution and quantification of free D-amino acids in serum and urine by comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry. AB - The potential of comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC*GC-TOFMS) in the quantitative analysis of amino acid enantiomers (AAEs) as their methyl chloroformate (MCF) derivatives in physiological fluids was investigated. Of the two column sets tested, the combination of an Rt-gammaDEXsa chiral column with a polar ZB-AAA column provided superior selectivity. Twenty AAEs were baseline resolved including L-Leu and D Ile, which had failed separation by one-dimensional chiral GC-quadrupole-MS (GC qMS). Lower limits of quantification (LLOQ) were in the range of 0.03-2 MUM. Reproducibility of the analysis of a serum specimen in octaplicate ranged from 1.3 to 16.6%. The GC*GC-TOFMS method was validated by analyzing AAEs in 48 urine and 43 serum specimens, respectively, and by comparing the results with data obtained by a previously validated GC-qMS method. Mean recoveries ranged from 78.4% for D-Leu to 116.4% for D-Pro in urine and 72.2% for L-Thr to 129.4% for L Ile in serum. The method was applied to the comparison of AAE serum levels in patients suffering from liver cirrhosis to a control group. Significantly increased D-AA concentrations were found for the patient group, whereas L-AA levels were slightly decreased. PMID- 21641601 TI - Determination of neonicotinoid pesticides residues in agricultural samples by solid-phase extraction combined with liquid chromatography-tandem mass spectrometry. AB - This work reports a new sensitive multi-residue liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for detection, confirmation and quantification of six neonicotinoid pesticides (dinotefuran, thiamethoxam, clothiandin, imidacloprid, acetamiprid and thiacloprid) in agricultural samples (chestnut, shallot, ginger and tea). Activated carbon and HLB solid-phase extraction cartridges were used for cleaning up the extracts. Analysis is performed by LC MS/MS operated in the multiple reaction monitoring (MRM) mode, acquiring two specific precursor-product ion transitions per target compound. Quantification was carried by the internal standard method with D(4)-labeled imidacloprid. The method showed excellent linearity (R(2)>=0.9991) and precision (relative standard deviation, RSD<=8.6%) for all compounds. Limits of quantification (LOQs) were 0.01 mg kg(-1) for chestnut, shallot, ginger sample and 0.02 mg kg(-1) for tea sample. The average recoveries, measured at three concentrations levels (0.01 mg kg(-1), 0.02 mg kg(-1) and 0.1 mg kg(-1) for chestnut, shallot, ginger sample, 0.02 mg kg(-1), 0.04 mg kg(-1) and 0.2 mg kg(-1) for tea sample), were in the range 82.1-108.5%. The method was satisfactorily validated for the analysis of 150 agricultural samples (chestnut, shallot, ginger and tea). Imidacloprid and acetamiprid were detected at concentration levels ranging from 0.05 to 3.6 mg kg( 1). PMID- 21641602 TI - Metabolite profiling on apple volatile content based on solid phase microextraction and gas-chromatography time of flight mass spectrometry. AB - A headspace SPME GC-TOF-MS method was developed for the acquisition of metabolite profiles of apple volatiles. As a first step, an experimental design was applied to find out the most appropriate conditions for the extraction of apple volatile compounds by SPME. The selected SPME method was applied in profiling of four different apple varieties by GC-EI-TOF-MS. Full scan GC-MS data were processed by MarkerLynx software for peak picking, normalisation, alignment and feature extraction. Advanced chemometric/statistical techniques (PCA and PLS-DA) were used to explore data and extract useful information. Characteristic markers of each variety were successively identified using the NIST library thus providing useful information for variety classification. The developed HS-SPME sampling method is fully automated and proved useful in obtaining the fingerprint of the volatile content of the fruit. The described analytical protocol can aid in further studies of the apple metabolome. PMID- 21641603 TI - Evaluation and single laboratory validation of an on-line turbulent flow extraction tandem mass spectrometry method for melamine in infant formula. AB - This report presents the single-laboratory validation of a method for the determination of melamine in dairy-based products using on-line turbulent flow extraction-tandem mass spectrometry. Liquid or powder test portions were dissolved in water, enriched with (13)C(3)(15)N(3)-Melamine internal standard, followed by protein precipitation and withdrawal of an aliquot for analysis. The turbulent flow method was validated by analyses of liquid and powdered proficiency test portions containing up to 10mg/kg melamine. Accuracy of results ranged from 96 to 106% of the assigned values for the 6 proficiency test portions tested with relative standard deviations of 4-8%. Apparent recoveries based on addition of amino-(15)N(3)-Melamine to prepared test portions were between 98 and 114%. Based on the repeat analysis of a known blank sample the limit of detection and limit of quantification were determined to be 27 and 87 MUg/kg, respectively. Additionally, this report demonstrates that turbulent flow chromatography is significantly faster than traditional LC-MS, with sample analysis times of less than 2 min. PMID- 21641604 TI - Comparison of gas chromatographic hyphenated techniques for mercury speciation analysis. AB - In this study, we evaluate advantages and disadvantages of three hyphenated techniques for mercury speciation analysis in different sample matrices using gas chromatography (GC) with mass spectrometry (GC-MS), inductively coupled plasma mass spectrometry (GC-ICP-MS) and pyrolysis atomic fluorescence (GC-pyro-AFS) detection. Aqueous ethylation with NaBEt(4) was required in all cases. All systems were validated with respect to precision, with repeatability and reproducibility <5% RSD, confirmed by the Snedecor F-test. All methods proved to be robust according to a Plackett-Burnham design for 7 factors and 15 experiments, and calculations were carried out using the procedures described by Youden and Steiner. In order to evaluate accuracy, certified reference materials (DORM-2 and DOLT-3) were analyzed after closed-vessel microwave extraction with tetramethylammonium hydroxide (TMAH). No statistically significant differences were found to the certified values (p=0.05). The suitability for water samples analysis with different organic matter and chloride contents was evaluated by recovery experiments in synthetic spiked waters. Absolute detection and quantification limits were in the range of 2-6 pg for GC-pyro-AFS, 1-4 pg for GC MS, with 0.05-0.21 pg for GC-ICP-MS showing the best limits of detection for the three systems employed. However, all systems are sufficiently sensitive for mercury speciation in environmental samples, with GC-MS and GC-ICP-MS offering isotope analysis capabilities for the use of species-specific isotope dilution analysis, and GC-pyro-AFS being the most cost effective alternative. PMID- 21641605 TI - Characterization of galactooligosaccharides derived from lactulose. AB - Galactooligosaccharides are non-digestible carbohydrates with potential ability to modulate selectively the intestinal microbiota. In this work, a detailed characterization of oligosaccharides obtained by transgalactosylation reactions of the prebiotic lactulose, by using beta-galactosidases of different fungal origin (Aspergillus oryzae, Aspergillus aculeatus and Kluveromyces lactis), is reported. Oligosaccharides of degree of polymerization (DP) up to 6 were detected and quantified by HPLC-ESI MS from a complex mixture produced by transgalactosylation reaction with A. oryzae (GOSLuAo), whereas only carbohydrates up to DP4 and DP5 were found for those obtained from the reaction with beta-galactosidases from K. lactis (GOSLuKl) and A. aculeatus (GOSLuAa), respectively. Disaccharides (galactosyl-galactoses and galactosyl-fructoses) and trisaccharides were characterised in the three mixtures by GC-MS as their trimethylsilyl oximes. Galactosyl- and digalactosyl-glycerols were produced during the transgalactosylation reaction of lactulose with beta-galactosidases from A. aculeatus and K. lactis, due to the presence of glycerol as enzyme stabiliser. PMID- 21641606 TI - Analytical methodology for multidimensional size/branch-length distributions for branched glucose polymers using off-line 2-dimensional size-exclusion chromatography and enzymatic treatment. AB - Instrumental and procedural optimizations are developed for a new method to obtain 2-dimensional distributions for branched homopolymers based on size and branching. The method uses 2-dimensional off-line size-exclusion chromatography (SEC) and chemical debranching, in this case using debranching enzymes on branched glucose polymers. This treatment, first presented for the 2D weight and number distributions of starch [F. Vilaplana, R.G. Gilbert, Macromolecules 43 (2010) 7321] is applied here to give 2D weight distributions as functions of size and branch length for glycogen, amylose, and native starch containing both amylopectin and amylose. Completely dissolved polysaccharides are first fractionated by size (hydrodynamic volume) using preparative SEC; the collected fractions are then debranched by addition of isoamylase, and the size distributions of the corresponding branches are analyzed for each fraction using analytical SEC with differential refractive index and with multiple-angle laser light scattering detection. Operational parameters have been optimized to provide sufficient separation resolution for each dimension (size of the whole branched macromolecule and size of the resulting branches after debranching) and to minimize degradation. These 2-dimensional distributions bring out new features in the structure of these branched polysaccharides, and offer a useful tool to elucidate relations between biosynthesis, structure, and properties. PMID- 21641607 TI - Characterization of mixed non-ionic surfactants n-octyl-beta-D-thioglucoside and octaethylene-glycol monododecyl ether: micellization and microstructure. AB - Mixed micelles of n-octyl-beta-D-thioglucoside (OTG) and octaethylene-glycol monododecyl ether (C(12)E(8)), two non-ionic surfactants belonging to the alkyl glucosides and polyoxyethylene alkyl ether families, respectively, were investigated by using light scattering and fluorescence probe techniques. From the determination of the critical micelle concentration (cmc), by the well established pyrene 1:3 ratio method, it was found that the mixed system behaves ideally, the micellization process being clearly controlled by the ethoxylated surfactant. The micellar hydrodynamic radius as a function of temperature, composition and concentration was obtained by dynamic light scattering measurements. It was observed that the micellar size increases with temperature, this growth being more pronounced as the relative proportion of the ethoxylated surfactant was increased. The behavior of the micellar size with the total surfactant concentration was also found to be dependent on temperature and composition. The clouding temperature, characteristic of the ethoxylated surfactants, was increased with the addition of the sugar surfactant. Lastly, possible structural changes in the micellar palisade layer were examined by steady-state fluorescence anisotropy in conjunction with time-resolved fluorescence studies with the hydrophobic probe coumarin 6 (C6). The obtained results indicate that the participation of the ethoxylated surfactant induces a slightly more polar palisade layer, whereas the probe carries out a faster rotational reorientation as a result of a less compact environment. All these observations were attributed to the different structure of the head groups of both surfactants and, as a consequence, to their different hydration. PMID- 21641608 TI - Novel ultrasonic-modified MnOx/TiO2 for low-temperature selective catalytic reduction (SCR) of NO with ammonia. AB - A novel ultrasonic-modified MnO(x)/TiO(2) catalyst was prepared and compared with two different kinds of MnO(x)/TiO(2) catalysts in the process of low-temperature selective catalytic reduction of NO with NH(3). The physicochemical properties of the catalysts were studied by using various characterization techniques, such as Brunauer-Emmett-Teller (BET) surface measurement, X-ray diffraction (XRD), high resolution transmission electron microscope (HRTEM), and in situ Fourier transform infrared spectroscopy (in situ FT-IR). The ultrasonic-modified process introduced ultrasound in the solution impregnation step of traditional impregnation method for MnO(x)/TiO(2) catalyst preparation. In this study, ultrasonic process significantly improved the dispersion behavior and surface acid property of manganese oxide on TiO(2) as well as the catalytic activity, especially at temperature below 120 degrees C. The NO conversion could reach 90% at 100 degrees C. For the novel ultrasonic-modified catalyst, the combination analysis of XRD and HRTEM confirmed that manganese oxide was in a highly dispersed state and Ti and Mn had strong interaction. Furthermore, in situ FT-IR studies revealed that there were significant amounts of Lewis acidity and high Mn atom concentration on the surface of the novel catalysts. PMID- 21641609 TI - Surface active ionic liquids: study of the micellar properties of 1-(1-alkyl)-3 methylimidazolium chlorides and comparison with structurally related surfactants. AB - The impetus for the increasing interest in studying surface active ionic liquids (SAILs; ionic liquids with long-chain "tails") is the enormous potential for their applications, e.g., in nanotechnology and biomedicine. The progress in these fields rests on understanding the relationship between surfactant structure and solution properties, hence applications. This need has prompted us to extend our previous study on 1-(1-hexadecyl)-3-methylimidazolium chloride to 1-(1-alkyl) 3-methylimidazolium chlorides, with alkyl chains containing 10, 12, and 14 carbons. In addition to investigating relevant micellar properties, we have compared the solution properties of the imidazolium-based surfactants with: 1-(1 alkyl)pyridinium chlorides, and benzyl (2-acylaminoethyl)dimethylammonium chlorides. The former series carries a heterocyclic ring head-group, but does not possess a hydrogen that is as acidic as H2 of the imidazolium ring. The latter series carries an aromatic ring, a quaternary nitrogen and (a hydrogen-bond forming) amide group. The properties of the imidazolium and pyridinium surfactants were determined in the temperature range from 15 to 75 degrees C. The techniques employed were conductivity, isothermal titration calorimetry, and static light scattering. The results showed the important effects of the interactions in the interfacial region on the micellar properties over the temperature range studied. PMID- 21641610 TI - How to obtain a well-spread monolayer of lysozyme at the air/water interfaces. AB - The purpose of this study is to define the conditions required to obtain a complete spreading of the lysozyme monolayer at the A/W interface. To this end, using Trurnit's method, the influence of the ionic strength of the substrate, the elapsed time between the spreading of the monolayer and the beginning of its compression, and the number of lysozyme molecules spread at the interface was studied. The results obtained show that the lysozyme spreading is conditioned by the unfolding of amino acid chains which form part of its structure, so that such unfolding is hindered, either because of an excessive accumulation of lysozyme molecules on the substrate surface or because the waiting time necessary to get this unfolding is not long enough, regardless of the number of spread molecules. The complete unfolding of lysozyme, which involves the loss of tertiary and secondary structures, has been obtained in this work under certain conditions: (1) using a substrate with a high saline concentration (NaCl 3-3.5 M) and at a pH value close to the isoelectric point of lysozyme (pI=11) in order to reduce the solubility and the electrical potential of the protein, (2) increasing the elapsed time between the spreading of the protein and the beginning of its compression to 3h in order to get a complete unfolding of lysozyme, which is a very slow process, (3) using an initial spreading area of 2.1m(2)/mg or larger (equivalent to a protein surface concentration of less than 2*10(12) molec/cm(2)) to reduce the accumulation of protein molecules at the surface, thus facilitating their unfolding, and (4) using Trurnit's spreading method, although for the spreading of this protein was found to be insufficiently significant. PMID- 21641611 TI - The bis(p-sulfonatophenyl)phenylphosphine-assisted synthesis and phase transfer of ultrafine gold nanoclusters. AB - Gold nanoclusters (Au NCs) have attracted intensive attention for their molecular like properties such as luminescence and unique charging behavior. A facile route has been developed for the preparation of ultrafine Au NCs at room temperature. Bis(p-sulfonatophenyl)phenylphosphine dihydrate dipotassium was used to stabilize the Au NCs obtained by NaBH(4) reduction of HAuCl(4) at pH of ~12 and facilitated the phase transfer of Au NCs from aqueous phase to an organic medium based on electrostatic interaction. From the analyses of TEM, HRTEM, and XRD patterns, the formation of fcc structured Au NCs dominated by {200} facets was identified. The Au NCs transferred into toluene could be used as seeds for the formation of core shell Au@Ag(2)S nanoparticles, providing for a promising strategy for the metal doping in semiconductor nanocrystals. PMID- 21641612 TI - Fabrication of block copolymer brushes on hollow sphere surface via reverse iodine transfer polymerization. AB - The block copolymer brushes grafted from hollow sphere surface via reverse iodine transfer polymerization (RITP) were investigated in this work. A sufficient amount of azo initiator was introduced onto hollow sphere surface firstly. Then the monomer methyl methacrylate (MMA) was polymerized via surface-initiated reverse iodine transfer polymerization (RITP) using azo group modified hollow sphere as initiator. The microstructure of the samples was characterized by FT IR, (1)H NMR, respectively. Results indicated that the poly(methyl methacrylate) (PMMA) with end functionality of alkyl iodine group had grafted from hollow sphere surface. TEM observations showed that the average diameter of hollow core was central at 1.3-1.4 MUm and the average wall thickness increased from 103 nm to 138 nm and 172 nm after grafting polymerization of MMA and Tb complex, respectively. The closely linear plots of molecular weight (M(n)) versus conversion, linear kinetic plots for the free polymer formed in solution and the ability to extend the chains by sequential addition of monomer indicated that the RITP was a controlled process with a "living" characteristic. PMID- 21641613 TI - Pb(II) and Zn(II) adsorption onto Na- and Ca-montmorillonites in acetic acid/acetate medium: experimental approach and geochemical modeling. AB - Smectites are usually used as a clay barrier at the bottom of subsurface waste landfills due to their low permeability and their capacity to retain pollutants. The Na- and Ca-saturated SWy2 montmorillonites were interacted with initial Zn(NO(3))(2) or Pb(NO(3))(2) concentrations ranging from 10(-6) to 10(-2)M with a solid/liquid ratio of 10 g L(-1) and using acetic acid/acetate as buffer at pH 5 in order to reproduce a biodegradable leachate of a young landfill. These experiments revealed that Zn and Pb sorption onto Na-SWy2 is higher than that onto Ca-SWy2 in the whole range of concentrations. Metal retention into both montmorillonites increases with the decrease in acetic acid/acetate concentration. The two-site protolysis model with no electrostatic term (2SPNE model) was used to model these experiments. As the experimental data of Zn sorption were well fitted, this model was validated and has been improved by taking into account the metal-acetate complexation in solution. In order to validate the model for Pb sorption, new selectivity coefficients have been determined, namely logK(c)(PbNa)=0.5 for Na-montmorillonite and logK(c)(PbCa)=0.3 for Ca-montmorillonite. PMID- 21641614 TI - To NEC or not to NEC: routine use of probiotics for premature infants. PMID- 21641615 TI - Birth prevalence rates of newborn screening disorders in relation to screening practices in the United States. AB - OBJECTIVE: To examine the associations between the first-tier-screening laboratory methods and criteria and the birth prevalence of congenital adrenal hyperplasia (CAH), phenylketonuria (PKU), and the sickle hemoglobinopathies occurring in the United States between 1991 and 2000. STUDY DESIGN: By using validated data from the National Newborn Screening and Genetics Resource Center, we fit Poisson regression models with laboratory methods and criteria used in every year for each state for each disorder. We also examined whether there was an overall change in birth prevalence over the decade and whether there was an effect resulting from obligatory second screenings. RESULTS: There were no associations among any of the factors and the birth prevalence of PKU in this decade. Use of the enzyme-linked immunosorbent assay was more likely than any other laboratory method to identify cases of CAH (OR 1.16; 95% CI 1.04-1.30), but no other factors were associated with this disorder. None of the factors examined were associated with the birth prevalence rates of any of the sickle hemoglobinopathies. CONCLUSION: There were no substantial changes in the birth prevalence rates of PKU, CAH, or the sickle hemoglobinopathies over the study period despite rapid changes in technology. PMID- 21641616 TI - In healthy young and elderly adults, hypothalamic-pituitary-adrenocortical axis reactivity (HPA AR) varies with increasing pharmacological challenge and with age, but not with gender. AB - BACKGROUND: Hypothalamic-pituitary-adrenocortical axis reactivity (HPA AR) is the key indicator of the psychophysiological response to stress. The HPA AR may vary with age and gender. To investigate these factors concurrently, the aims of the present study were to observe HPA AR (plasma ACTH and plasma cortisol) in response to a pharmacological challenge (dexamethasone/corticotropin releasing hormone test: DEX/CRH-test) and as a function of age and gender. METHOD: 19 young (10 females and 9 males; mean age = 24.05 years) and 23 elderly (11 females and 12 males; mean age = 71.61 years) healthy volunteers took part in the study. To assess HPA AR, participants underwent the combined DEX/CRH test applied with the following DEX doses: 0.75, 1.5, and 3.0 mg, respectively. RESULTS: A dose dependent response was observed in young adult participants, but not in elderly participants. With increasing DEX doses, ACTH and cortisol values decreased in young adult participants, while the decrease was blunted among elderly compared to young adult participants. No differences were observed for gender. CONCLUSIONS: Results point to diminished HPA axis sensitivity as an effect of normal aging, irrespective of gender. Therefore, altered HPA regulation in old age should be taken into account for developing new therapeutic approaches acting on the HPA axis and its receptor mechanisms. PMID- 21641617 TI - Systemic toxicity in early-stage mood disorders. PMID- 21641618 TI - Proteasome inhibition induces alpha-synuclein SUMOylation and aggregate formation. AB - Parkinson's disease (PD) and Dementia with Lewy Bodies (DLB) are characterized pathologically by intraneuronal inclusions called Lewy bodies (LBs) and Lewy neurites. A major component of these inclusions is the protein alpha-synuclein, which is natively unfolded but forms oligomers and insoluble fibrillar aggregates under pathological conditions. Although alpha-synuclein is known to undergo several posttranslational modifications, the contribution of SUMOylation to alpha synuclein aggregation and the pathogenesis of alpha-synucleinopathies have not been elucidated. Here, we provide evidence that aggregates and inclusions formed as a result of impaired proteasome activity contain SUMOylated alpha-synuclein. Additionally, SUMO1 is present in the halo of LBs colocalizing with alpha synuclein in the brains of PD and DLB patients. Interestingly, SUMOylation does not affect the ubiquitination of alpha-synuclein. These findings suggest that proteasomal dysfunction results in the accumulation of SUMOylated alpha-synuclein and subsequently its aggregation, pointing to the contribution of this posttranslational modification to the pathogenesis of inclusion formation in alpha-synucleinopathies. PMID- 21641619 TI - CD24 as a genetic modifier of disease progression in multiple sclerosis in Argentinean patients. AB - INTRODUCTION: Previous reports have shown that CD24 gene polymorphisms have an important role in the risk of development and progression of multiple sclerosis (MS). OBJECTIVE: To investigate the association between P226 polymorphisms (T/C), P1056 (A/G), P1527 (TG/del) and P1626 (A/G) of the CD24 gene and MS, comparing allele and genotype frequencies of patients versus controls. MATERIALS AND METHODS: We analyzed DNA samples from 102 MS patients and from 205 unrelated healthy individuals. DNA was extracted from peripheral blood and polymorphic regions were amplified by nested PCR. Genotyping was performed by restriction fragments length polymorphisms. Time from disease onset to reach EDSS 6 and time to conversion to secondary progressive phase (SP) were used as variables of survival as well as percentage of patients that reached those endpoints. We used the log Rank test for data comparison (significant p<=0.05). RESULTS: We found no differences between cases and controls in frequency of polymorphisms at the CD24 gene. 44.6% of patients with the AA genotype (P1626) reached an EDSS 6 vs 16% of patients with other genotypes (p<0.001, HR 3.2, 95% CI 1.4 to 7.4). 45.8% of patients with the AA genotype reached SPMS vs 16.7% without this genotype (p<0.001, HR 3.4, 95% CI 1.5 to 7.8). CONCLUSIONS: This study showed a strong association between the presence of AA genotype in the 1626 polymorphism of the CD24 gene and the risk of disease progression in MS patients. PMID- 21641620 TI - New threats of an old enemy: the distribution of the shipworm Teredo navalis L. (Bivalvia: Teredinidae) related to climate change in the Port of Rotterdam area, the Netherlands. AB - The effects of four climate change scenarios for the Netherlands on the distribution of the shipworm upstream of the Rhine-Meuse estuary are described. Global warming will cause dry and warmer summers and decreased river discharges. This will extend the salinity gradient upstream in summer and fall and may lead to attacks on wooden structures by the shipworm. Scenarios including one or two degree temperature increases by 2050 compared to 1990 with a weak change in the air circulation over Europe will lead to an increased chance of shipworm damage upstream from once in 36 years to once in 27 or 22 years, respectively; however, under a strong change in air circulation, the chance of shipworm damage increases to once in 6 or 3 years, respectively. The upstream expansion of the distribution of the shipworm will also be manifested in other northwest European estuaries and will be even stronger in southern Europe. PMID- 21641621 TI - Origin of the mass mortality of the flathead grey mullet (Mugil cephalus) in the Tanshui River, northern Taiwan, as indicated by otolith elemental signatures. AB - A massive fish kill of approximately 50 tons of flathead grey mullet (Mugil cephalus) occurred in August 2006 in the Tanshui River, northern Taiwan. To clarify the origin of these mullets, 42 individuals were randomly collected to determine their ages by reading otolith annuli and to measure the trace elements in the otolith. The total lengths of the fish averaged 32.7+/-4.4 cm, and most of the individuals were 2 years old, indicating that the mullets in the mass fish kill event were in the immature and growth stages. The Sr/Ca and Ba/Ca ratios of a subsample of 14 otoliths showed that half of the fish killed were estuarine residents, and the rest originated from marine and freshwater habitats. The multiple causes involved in this fish kill implied that additional unknown factors influenced the mullet populations on a large scale and induced them to move quickly and gather at this estuary. PMID- 21641622 TI - Stable isotopes as a useful tool for revealing the environmental fate and trophic effect of open-sea-cage fish farm wastes on marine benthic organisms with different feeding guilds. AB - Environmental fate of fish farm wastes (FFW) released from an open-sea-cage farm at Kat O, Hong Kong was examined by measuring carbon and nitrogen stable isotope (SI) ratios in selected benthic organisms collected along a 2000 m transect from the farm. Our results showed that FFW significantly influenced the energy utilization profile of consumers near the fish farm. Although nitrogen enrichment effect on delta15N was anticipated in biota near the farm, the predicted patterns did not consistently occur in all feeding guilds. Two species of suspension feeders, which relied on naturally delta15N-depleted sources, were delta15N enriched near the fish farm. In contrast, both species of benthic grazer and deposit-feeder, which relied on naturally delta15N-enriched algal sources, were delta15N-depleted under the influence of FFW. The SI signatures of biota can, therefore, serve as feasible biomarkers for FFW discharges only when the trophic structure of the receiving environment is fully elucidated. PMID- 21641623 TI - Flavonoids from the leaves and stems of Dodonaea polyandra: a Northern Kaanju medicinal plant. AB - Three prenylated flavonoids 5,7,4'-trihydroxy-3'(3-methylbut-2-enyl)-3-methoxy flavone, 5,7-dihydroxy-3'(3-methylbut-2-enyl)-3,4'-dimethoxy flavone and 5,7,4' trihydroxy-3',5'(3-methylbuyt-2-enyl)-3-methoxy flavone together with three other known flavonoids were isolated from the medicinal plant Dodonaea polyandra. The plant is used in the traditional medicine system of Northern Kaanju people of Cape York Peninsula, Queensland, Australia. The extracts studied have previously been found to possess anti-inflammatory activity. Successive fractionation of leaf and stem extracts by column and high performance liquid chromatography led to the isolation of these compounds. Their structures were determined using a number of spectroscopic techniques including 1D and 2D NMR and high resolution mass spectroscopy. The structural elucidation is reported herein accompanied by full 1H and 13C NMR spectroscopic data. Spectroscopic data of known compounds was in agreement with that previously reported in literature. PMID- 21641624 TI - [Impact of mammographic breast density on computer-assisted detection (CAD) in a breast imaging department]. AB - OBJECTIVE: To evaluate whether breast density influences the sensitivity of a computer-assisted detection (CAD) system for the detection of breast cancer. MATERIAL AND METHODS: We prospectively studied 8750 digital mammograms with an associated CAD system. We used BI-RADS criteria to classify breast density. We calculated the overall sensitivity of the radiologist and of the CAD system, as well as the sensitivity for each projection and type of finding in relation to the mammographic density of the breast. Finally, we analyzed the interval carcinomas. We used SPSS 11 for all statistical analyses. RESULTS: The overall sensitivity of the CAD system was 88.5% (95% CI: 83.2-92.7%), and the overall sensitivity of the radiologist was 93.5% (95% CI: 84.4%-95.5%). The sensitivity of the craniocaudal view was 81.6% (95% CI: 76.5-90.7%) vs 76.5% (95% CI: 69.3 89.3%) for the mediolateral oblique view. The sensitivity for microcalcifications was 98.6% (95% CI: 96.5-99.7%), and the sensitivity for masses 83.4% (95% CI: 81.2-91.7%). We detected discrepancies smaller than 20% both for microcalcifications present in the four types of densities and for masses with densities 1 and 2. In masses with density 3 the discrepancy was 20.8% and in those with density 4 it was 55%. The CAD system failed to mark only 9.1% (9/94) of the cancers presenting as masses. Half of the interval carcinomas were found in type 4 density and 75% manifested as masses, asymmetries, and distortions. The CAD system had marked 35.7% of the carcinomas. CONCLUSIONS: The craniocaudal view was more sensitive, although this difference was not statistically significant. The sensitivity of CAD was high for microcalcifications in all four density types; however, CAD's sensitivity for masses was low in density types 3 and 4. The CAD system only failed to mark 9.1% of the cancers presenting as masses but was not sensitive for the other two radiological findings included in this marking. Half of the interval carcinomas occurred in type 4 densities and 35.7% had been marked by the CAD system. PMID- 21641625 TI - [Joubert syndrome: findings at conventional magnetic resonance image and at diffusion tensor imaging]. AB - We present the case of a 20-year-old man previously diagnosed with cerebral palsy in whom a developmental disorder was detected at physical examination. After cranial CT and conventional cranial MRI, we diagnosed Joubert syndrome. We completed the study with diffusion tensor imaging and tractography. This article reviews the imaging findings for Joubert syndrome and the relevant literature. PMID- 21641626 TI - [Abdominal ultrasonography versus abdominal CT in the follow-up of testicular germ cell tumors]. AB - INTRODUCTION: Testicular germ cell tumors are common in young men. There is a wide consensus regarding the key points in their diagnosis and treatment, although some elements, including the best approach to follow-up, are being reviewed and revised. We present a statistical study that uses tools for the evaluation of diagnostic tests to compare the usefulness of abdominal ultrasonography (US) in comparison with that of abdominal CT, taken as the gold standard, in the detection of liver metastases and retroperitoneal adenopathies in patients with testicular germ cell tumors. MATERIAL AND METHODS: We analyzed a total of 308 diagnostic tests (154 CT studies and 154 US studies) from 59 patients with at least one year's follow-up at our institution. Patients underwent abdominal US before abdominal CT following a standard protocol. RESULTS: Compared to the gold standard, abdominal US had 95% sensitivity, 92% specificity, 82% PPV, and 98% NPV. CONCLUSION: US is very sensitive and can be used in protocols for the follow-up of primary testicular tumors to rule out disease. PMID- 21641627 TI - Characterisation of the impact of coagulation and anaerobic bio-treatment on the removal of chromophores from molasses wastewater. AB - The performance of a coagulation sequence using aluminium chlorohydrate (ACH) and a low MW polydiallyldimethylammonium chloride (polyDADMAC), and ferric chloride, for decolourising a high-strength industrial molasses wastewater was compared at bench scale. At their optimum dosages, ACH/polyDADMAC gave higher colour removal than FeCl(3) (45% cf. 28%), whereas COD reduction was similar (~30%), indicating preferential removal of melanoidins (a major contributor to the colour) by ACH/polyDADMAC. Size exclusion chromatography and fluorescence excitation emission matrix spectrometry suggested that chromophoric Fe-organic complexes were formed during FeCl(3) treatment of the molasses wastewater, which appeared to compromise decolourisation efficiency. Anaerobic bio-treatment of the wastewater enhanced the coagulation efficiency markedly, with FeCl(3) achieving 94% colour and 96% COD removal, while ACH/polyDADMAC gave 70% and 56% removal, respectively. The improved decolourisation was attributed to the decrease in low MW organics (<500 Da) and biopolymers by the biological treatment, leading to reduced competition with melanoidins for interaction with coagulant/flocculant. For both the wastewater and the biologically treated wastewater, ACH/polyDADMAC treatment gave flocs with markedly better settling properties compared with FeCl(3). PMID- 21641628 TI - Detection of pharmaceutically active compounds in the rivers and tap water of the Madrid Region (Spain) and potential ecotoxicological risk. AB - Concentrations of pharmaceutically active compounds (PhACs) in the order of ng L( 1) to MUg L(-1) have been reported worldwide in waste, fluvial and even drinking water, raising concern about the efficacy of the currently employed waste water treatments in the elimination of this kind of compounds. Despite ranking 29th in terms of population, Spain is currently the 8th country on pharmaceutical prescription with an expense of 14*10(9) euros in 2008. In this context, the aim of this study was to determine the presence of 33 pharmaceutically active compounds in specific points of the main rivers of the Madrid Region (MR) as well as tap water samples from the metropolitan area of Madrid. Additionally, a screening level risk characterization by means of the Hazard Quotient (HQ) method was applied. A total of 25 pharmaceutical compounds and metabolites were detected in the 10 sampling points downstream the outlet of the major STPs of the MR. The highest concentrations were detected for the anticonvulsant carbamazepine and the stimulant caffeine. Concentrations for most of the analyzed compounds exceed levels previously reported in the literature. Moreover, we report the highest concentration of the cytostatic ifosfamide, detected for the first time in Spain in surface water. Preliminary risk characterization shows that a total of 16 compounds represent at least a low potential hazard based on their scored HQs, with five of them present in a concentration that exceeds the predicted no effect concentration (PNEC). Toxic Units calculation indicates that for all the selected sampling points high hazard is anticipated from the presence of the analyzed compounds in the measured concentrations (TUs>10). Caffeine and cotinine were detected in all (10) the analyzed tap water samples. Carbamazepine and nicotine were detected in six and venlafaxine in two samples. No studies venlafaxine in drinking water have been reported. These results clearly pinpoint the need for water quality monitoring and research in urban rivers, as well as the need for improved water treatment techniques able to eliminate this kind of compounds from the effluent waters as well as from drinking water sources. PMID- 21641629 TI - Field performance evaluation of a newly developed PM2.5 sampler at IIT Kanpur. AB - In order to meet the challenges of growing air pollution for a developing nation and to measure the ambient fine particles (PM2.5, particles having aerodynamic diameter less than 2.5 MUm) on routine basis an air sampler was designed, developed and evaluated in the field. The impactor removes particles greater than 2.5 MUm from the air stream via impacting them onto a vacuum grease substrate and finer particles get eventually collected on a backup filter. Various impactor nozzles with conical geometry were designed based on the published theoretical design equations. A detail parametric investigation was carried out which resulted in the optimum impactor nozzle design. For this exercise, a novel dry aerosol generator was employed in addition to the well known time-of-flight instrument, APS (Aerodynamic Particle Sizer, Model 3021, TSI Inc.). The average particle losses for the impactor nozzle as well as the sampler body were below 10% and the overall pressure drop (including a backup 47 mm filter) through the PM2.5 sampler was only 2 in. of H2O. This developed PM2.5 sampler operates at a flow rate of 15 LPM. Field performance of this sampler was evaluated through co located sampling with a high volume PM2.5 reference sampler (HVS, GEM-BLI Model 2360, Tisch Environment Instrument) within the IIT Kanpur campus. The sampling period was 10 h long and it was carried out on six different days. The entire sets of filters were analyzed gravimetrically followed by their chemical analysis for elemental and anionic analyses. The particle mass, elemental, and anionic concentrations obtained with this newly developed PM2.5 sampler as well as those from the reference HVS sampler showed moderate to good correlation. PMID- 21641630 TI - Factors controlling mobility of 127I and 129I species in an acidic groundwater plume at the Savannah River Site. AB - In order to quantify changes in iodine speciation and to assess factors controlling the distribution and mobility of iodine at an iodine-129 ((129)I) contaminated site located at the U.S. Department of Energy's Savannah River Site (SRS), spatial distributions and transformation of (129)I and stable iodine ((127)I) species in groundwater were investigated along a gradient in redox potential (654 to 360 mV), organic carbon concentration (5 to 60 MUmol L(-1)), and pH (pH 3.2 to 6.8). Total (129)I concentration in groundwater was 8.6+/-2.8 Bq L(-1) immediately downstream of a former waste seepage basin (well FSB-95DR), and decreased with distance from the seepage basin. (127)I concentration decreased similarly to that of (129)I. Elevated concentrations of (127)I or (129)I were not detected in groundwater collected from wells located outside of the mixed waste plume of this area. At FSB-95DR, the majority (55-86%) of iodine existed as iodide for both (127)I and (129)I. Then, as the iodide move down gradient, some of it transformed into iodate and organo-iodine. Considering that iodate has a higher K(d) value than iodide, we hypothesize that the production of iodate in groundwater resulted in the removal of iodine from the groundwater and consequently decreased concentrations of (127)I and (129)I in downstream areas. Significant amounts of organo-iodine species (30-82% of the total iodine) were also observed at upstream wells, including those outside the mixed waste plume. Concentrations of groundwater iodide decreased at a faster rate than organo iodine along the transect from the seepage basin. We concluded that removal of iodine from the groundwater through the formation of high molecular weight organo iodine species is complicated by the release of other more mobile organo-iodine species in the groundwater. PMID- 21641631 TI - Size resolved ultrafine particles emission model--a continues size distribution approach. AB - A new parameterization for size resolved ultrafine particles (UFP) traffic emissions is proposed based on the results of PARTICULATES project (Samaras et al., 2005). It includes the emission factors from the Emission Inventory Guidebook (2006) (total number of particles, #/km/veh), the shape of the corresponding particle size distribution given in PARTICULATES and data for the traffic activity. The output of the model UFPEM (UltraFine Particle Emission Model) is a sum of continuous distributions of ultrafine particles emissions per vehicle type (passenger cars and heavy duty vehicles), fuel (petrol and diesel) and average speed representative for urban, rural and highway driving. The results from the parameterization are compared with measured total number of ultrafine particles and size distributions in a tunnel in Antwerp (Belgium). The measured UFP concentration over the entire campaign shows a close relation to the traffic activity. The modelled concentration is found to be lower than the measured in the campaign. The average emission factor from the measurement is 4.29E+14 #/km/veh whereas the calculated is around 30% lower. A comparison of emission factors with literature is done as well and in overall a good agreement is found. For the size distributions it is found that the measured distributions consist of three modes--Nucleation, Aitken and accumulation and most of the ultrafine particles belong to the Nucleation and the Aitken modes. The modelled Aitken mode (peak around 0.04-0.05 MUm) is found in a good agreement both as amplitude of the peak and the number of particles whereas the modelled Nucleation mode is shifted to smaller diameters and the peak is much lower that the observed. Time scale analysis shows that at 300 m in the tunnel coagulation and deposition are slow and therefore neglected. The UFPEM emission model can be used as a source term in dispersion models. PMID- 21641632 TI - Effect of injection of di- and tricyclic aromatic compounds on post-combustion formation of polychlorinated dibenzo-p-dioxins and dibenzofurans. AB - The formation of mono- to octachlorinated dibenzo-p-dioxins (PC1-8DD) and dibenzofurans (PC1-8DF) was studied using a model waste in a laboratory-scale combustion reactor with simultaneous collection of flue gas at three different temperatures (450 degrees C, 300 degrees C, and 200 degrees C) in the post combustion zone. To investigate the influence of chlorination reactions and the effects of carbon backbone-containing compounds present in the flue gases, five aromatic compounds were injected into the flue gas, namely dibenzofuran (DF), biphenyl (BP), naphthalene, phenanthrene and fluorene. The injection of DF induced a reduction in the concentration of PC3-5DD, but did not significantly influence the concentration of PCDF. A reduction in the concentration of PC3-5DD was also observed during the injection of fluorene, which is structurally very similar to DF. The injection of biphenyl, naphthalene and phenanthrene had less pronounced effects on the formation of PCDD and PCDF. A possible explanation of the observed changes during injection of DF and fluorene, based on homologue profiles and affected congeners, involves formation of radical species from fluorene and/or dibenzofuran. The fluorene radical is stabilized by the delocalization of electrons across the aromatic ring structure and has the propensity to react with highly abundant hydrogen chloride, whereas the molecular species would require reaction with Cl2 or chlorine radicals. PMID- 21641633 TI - Haemostatic and inflammatory markers are independently associated with myocardial infarction in men and women. AB - INTRODUCTION: Previous studies have shown that plasma levels of haemostatic and inflammatory markers are associated with risk of coronary heart disease (CHD). As haemostatic markers are also acute-phase reactants, it is not clear if their association with CHD is independent of inflammatory markers and established cardiovascular risk factors. MATERIALS AND METHODS: We used a prospective incident case-control study design nested in two cohorts from Sweden. Baseline measurements of a panel of cardiovascular risk factors and eight established markers of haemostasis or inflammation were assessed in 469 first-ever myocardial infarction (MI) cases and 895 matched controls. RESULTS: After adjustment for baseline values of established risk factors, von Willebrand factor appeared to have the strongest association with MI among the haemostatic markers assayed, with an odds ratio of 2.52 (95% CI, 1.72-3.67) for a comparison of individuals in extreme thirds of baseline levels. For a similar comparison, after adjustment for established risk factors and haemostatic markers, odds ratios for IL-6 and CRP were 1.67 (95% CI, 1.08-2.60) and 1.58 (95% CI, 1.03-2.41), respectively. The relative predictive ability of the individual markers over and above established risk factors was modest according to comparisons of Area under the Receiver Operating Characteristic (AUROC) curves. However, when all eight markers were combined in a single model, the AUROC curve was significantly increased to 0.820 (95% CI, 0.795-0.846) compared to 0.762 (95% CI, 0.732-0.791) for established risk factors only. CONCLUSIONS: These findings suggest that haemostasis and inflammation have at least partially separate roles in risk of myocardial infarction. PMID- 21641634 TI - Kinetic modeling sheds light on the mode of action of recombinant factor VIIa on thrombin generation. AB - INTRODUCTION: The therapeutic potential of a hemostatic agent can be assessed by investigating its effects on the quantitative parameters of thrombin generation. For recombinant activated factor VII (rFVIIa)--a promising hemostasis-inducing biologic--experimental studies addressing its effects on thrombin generation yielded disparate results. To elucidate the inherent ability of rFVIIa to modulate thrombin production, it is necessary to identify rFVIIa-induced effects that are compatible with the available biochemical knowledge about thrombin generation mechanisms. MATERIALS AND METHODS: The existing body of knowledge about coagulation biochemistry can be rigorously represented by a computational model that incorporates the known reactions and parameter values constituting the biochemical network. We used a thoroughly validated numerical model to generate activated factor VII (FVIIa) titration curves in the cases of normal blood composition, hemophilia A and B blood, blood lacking factor VII, blood lacking tissue factor pathway inhibitor, and diluted blood. We utilized the generated curves to perform systematic fold-change analyses for five quantitative parameters characterizing thrombin accumulation. RESULTS: The largest fold changes induced by increasing FVIIa concentration were observed for clotting time, thrombin peak time, and maximum slope of the thrombin curve. By contrast, thrombin peak height was much less affected by FVIIa titrations, and the area under the thrombin curve stayed practically unchanged. Comparisons with experimental data demonstrated that the computationally derived patterns can be observed in vitro. CONCLUSIONS: rFVIIa modulates thrombin generation primarily by accelerating the process, without significantly affecting the total amount of generated thrombin. PMID- 21641635 TI - A novel approach in allergen-specific immunotherapy: combination of sublingual and subcutaneous routes. AB - BACKGROUND: Subcutaneous allergen-specific immunotherapy (SIT) has an early onset of action, whereas repeated injections and safety concerns have limited its use in the pediatric age group. Meanwhile, the improved safety profile of the sublingual route has been accepted as an alternative despite its relatively late onset of action. OBJECTIVE: We sought to improve the efficacy and safety of SIT with a combination of the subcutaneous route in the build-up phase and sublingual maintenance in comparison with the sublingual or subcutaneous routes alone. METHODS: Fifty-one house dust mite-sensitized children with mild-to-moderate asthma were randomized into one of 4 groups to receive either (1) subcutaneous immunotherapy (SCIT), (2) sublingual immunotherapy (SLIT), (3) SCIT plus SLIT, or (4) pharmacotherapy. Clinical parameters were evaluated at baseline and months 1, 4, 12, and 18. Allergen-specific immunoglobulin levels and allergen-induced IL-5, IL-10, IL-13, IL-17, TGF-beta, and IFN-gamma levels were evaluated as well. RESULTS: In the SCIT and SCIT plus SLIT groups, the number of asthma attacks and inhaled corticosteroid dosage decreased compared with baseline values at the months 4, 12, and 18 but only at month 12 in the SLIT group. The improvement in visual analog scores for rhinitis was significant only in the SCIT plus SLIT group. Increases in the levels of regulatory and T(H)1 cytokines were observed both in the SCIT and SLIT groups, with some differences in dynamics. Antigen specific IgG(4) levels increased in the SCIT and SCIT plus SLIT groups but not in the SLIT group. Clinical symptom scores were correlated positively with IL-5 levels and negatively with antigen-specific IgG(4), IFN-gamma, and TGF-beta levels. CONCLUSION: Our novel regimen of immunotherapy, SCIT plus SLIT, appeared promising in that it successfully combined the advantages of the 2 alternatives: rapid onset and potency in SCIT and safety and avoidance of injections in SLIT. PMID- 21641637 TI - First-line treatment of advanced colorectal cancer. PMID- 21641638 TI - Biodegradation and in vivo biocompatibility of a degradable, polar/hydrophobic/ionic polyurethane for tissue engineering applications. AB - A degradable, polar/hydrophobic/ionic polyurethane (D-PHI) scaffold was optimized in in vitro studies to yield mechanical properties appropriate to replicate vascular graft tissue while eliciting a more wound-healing phenotype macrophage when compared to established materials. The objectives of this study were to characterize the biodegradation (in vitro and in vivo) and assess the in vivo biocompatibility of D-PHI, comparing it to a well-established, commercially available scaffold biomaterial, polylactic glycolic acid (PLGA), recognized as being degradable, non-cytotoxic, and showing good biocompatibility. PLGA and D PHI were formed into 6 mm diameter disk-shaped scaffolds (2 mm thick) of similar porosity (~82%) and implanted subcutaneously in rats. Both PLGA and D-PHI scaffolds were well-tolerated at the 7 d time point in vivo. In vitro D-PHI scaffolds degraded slowly (only 12 wt% in PBS in vitro after 120 d at 37 degrees C). In vivo, D-PHI scaffolds degraded at a more controlled rate (7 wt% loss over the acute 7 d implant phase and subsequently a linear profile of degradation leading to a 21 wt% mass loss by 100 d (chronic period)) than PLGA scaffolds which showed an initial more rapid degradation (14 wt% over 7 d), followed by minimal change between 7 and 30 d, and then a very rapid breakdown of the scaffold over the next 60 d. Histological examination of D-PHI scaffolds showed tissue ingrowth into the pores increased with time whereas PLGA scaffolds excluded cells/tissue from its porous structure as it degraded. The results of this study suggest that D-PHI has promising qualities for use as an elastomeric scaffold material for soft TE applications yielding well integrated tissue within the scaffold and a controlled rate of degradation stabilizing the form and shape of the implant. PMID- 21641639 TI - MRI monitoring of intratumoral drug delivery and prediction of the therapeutic effect with a multifunctional thermosensitive liposome. AB - Non-invasive in vivo imaging of drug distribution enables real-time monitoring and prediction of therapeutic responses to treatment. We have developed a thermosensitive liposomal formulation (HaT: Hyperthermia-activated-cytoToxic) consisting of DPPC and Brij78, a formulation that enhanced drug delivery compared to the lyso-lipid temperature sensitive liposomes (LTSL). Here we report the development of a multifunctional HaT liposome co-encapsulating Gd-DTPA (an MRI probe) and doxorubicin (DOX), which simultaneously releases and reports on drug delivery in a locally heated tumor. The temperature-dependent release profiles of DOX from HaT were closely related to the change in the MR T(1) relaxation time, in which DOX was 100% released at 40-42 degrees C in 3 min, accompanied by a 60% reduction in T(1). By T(1) relaxometry analysis, no Gd-DTPA leakage was detected in 30 min at 30-37 degrees C. In the in vivo study, DOX uptake in the tumor was quantitatively correlated with T(1) response (R(2) = 0.98) and the patterns of the T(1) image and the intratumoral DOX uptake were matched, in which both signals were predominantly detected in the highly perfused tumor periphery. Finally, the extent of T(1) relaxation enhancement in the heated tumor successfully predicted the antitumor efficacy in a standard pharmacological response model (R(2) = 0.98). PMID- 21641640 TI - Sex-specific differences in placental global gene expression in pregnancies complicated by asthma. AB - BACKGROUND: Chronic maternal asthma is associated with reduced growth of the female fetus and normal growth of the male fetus. The mechanisms that control the differential effects of maternal asthma on the fetus have not been fully elucidated but alterations in placental function may play a role. In the current study we have used microarray platform to examine fetal sex-specific global changes in placental gene expression in pregnancies complicated by asthma as compared to non-asthmatic subjects. METHODS: Placental RNA was extracted from 11 control subjects and 38 asthmatic subjects. Labeled cDNA was hybridized to an oligonucleotide chip with 1700 double spotted well-characterized human genes. Global gene expression data analysis and visualization were performed using the Binary Tree-Structured Vector Quantization (BTSVQ) software. Functional relationships of differentially expressed genes were assessed using protein protein interaction database I2D, network analysis and visualization software NAViGaTOR and Ingenuity Pathway Analysis software. RESULTS: Overall, 65 genes were found to be altered in placentae of pregnancies complicated by asthma. Of these, only 6 genes were altered in male placentae. There were 59 gene changes in female placentae of asthmatic mothers relative to control placentae. Some of the sex-specific genes were associated with growth, inflammation and immune pathways. CONCLUSION: There are sex-specific alterations in placental gene expression in the presence of maternal asthma. Given that many of the identified genes in the female placentae were associated with or involved in cellular growth and tissue development, these may contribute to the sexually dimorphic difference in fetal growth in response to maternal asthma. PMID- 21641636 TI - Addition of cetuximab to oxaliplatin-based first-line combination chemotherapy for treatment of advanced colorectal cancer: results of the randomised phase 3 MRC COIN trial. AB - BACKGROUND: In the Medical Research Council (MRC) COIN trial, the epidermal growth factor receptor (EGFR)-targeted antibody cetuximab was added to standard chemotherapy in first-line treatment of advanced colorectal cancer with the aim of assessing effect on overall survival. METHODS: In this randomised controlled trial, patients who were fit for but had not received previous chemotherapy for advanced colorectal cancer were randomly assigned to oxaliplatin and fluoropyrimidine chemotherapy (arm A), the same combination plus cetuximab (arm B), or intermittent chemotherapy (arm C). The choice of fluoropyrimidine therapy (capecitabine or infused fluouroracil plus leucovorin) was decided before randomisation. Randomisation was done centrally (via telephone) by the MRC Clinical Trials Unit using minimisation. Treatment allocation was not masked. The comparison of arms A and C is described in a companion paper. Here, we present the comparison of arm A and B, for which the primary outcome was overall survival in patients with KRAS wild-type tumours. Analysis was by intention to treat. Further analyses with respect to NRAS, BRAF, and EGFR status were done. The trial is registered, ISRCTN27286448. FINDINGS: 1630 patients were randomly assigned to treatment groups (815 to standard therapy and 815 to addition of cetuximab). Tumour samples from 1316 (81%) patients were used for somatic molecular analyses; 565 (43%) had KRAS mutations. In patients with KRAS wild-type tumours (arm A, n=367; arm B, n=362), overall survival did not differ between treatment groups (median survival 17.9 months [IQR 10.3-29.2] in the control group vs 17.0 months [9.4-30.1] in the cetuximab group; HR 1.04, 95% CI 0.87-1.23, p=0.67). Similarly, there was no effect on progression-free survival (8.6 months [IQR 5.0-12.5] in the control group vs 8.6 months [5.1-13.8] in the cetuximab group; HR 0.96, 0.82 1.12, p=0.60). Overall response rate increased from 57% (n=209) with chemotherapy alone to 64% (n=232) with addition of cetuximab (p=0.049). Grade 3 and higher skin and gastrointestinal toxic effects were increased with cetuximab (14 vs 114 and 67 vs 97 patients in the control group vs the cetuximab group with KRAS wild type tumours, respectively). Overall survival differs by somatic mutation status irrespective of treatment received: BRAF mutant, 8.8 months (IQR 4.5-27.4); KRAS mutant, 14.4 months (8.5-24.0); all wild-type, 20.1 months (11.5-31.7). INTERPRETATION: This trial has not confirmed a benefit of addition of cetuximab to oxaliplatin-based chemotherapy in first-line treatment of patients with advanced colorectal cancer. Cetuximab increases response rate, with no evidence of benefit in progression-free or overall survival in KRAS wild-type patients or even in patients selected by additional mutational analysis of their tumours. The use of cetuximab in combination with oxaliplatin and capecitabine in first-line chemotherapy in patients with widespread metastases cannot be recommended. FUNDING: Cancer Research UK, Cancer Research Wales, UK Medical Research Council, Merck KGgA. PMID- 21641641 TI - Differential effects of urinary and recombinant chorionic gonadotropin on oxidative stress responses in decidualizing human endometrial stromal cells. AB - Human chorionic gonadotropin (hCG) is one of the earliest signals secreted by the implanting embryo. In addition to its well-known luteotropic function in early pregnancy, hCG also acts directly on decidualizing endometrium. Recently, we demonstrated that recombinant hCG (rhCG) prevented apoptosis in decidualizing human endometrial stromal cells (HESCs) exposed to oxidative stress. Two hCG preparations are widely used clinically: rhCG, produced by recombinant DNA technology, and urinay hCG (uhCG), extracted from urine of post-menopausal women. However, an analysis of the direct effects of rhCG and uhCG on the decidual phenotype of HESCs has not yet been done. In this study, we investigated the effects of uhCG and rhCG on the morphological and functional profiles of decidualizing HESCs. We demonstrate that neither rhCG nor uhCG alter the morphological appearance of the decidual HESC cultures, although rhCG but not uhCG attenuated prolactin expression, a major decidual marker protein. Moreover, rhCG, but not uhCG, protected decidualizing HESCs from oxidative cell death, mediated at least in part by two major mechanisms. First, rhCG, but not uhCG, enhances the expression of manganese superoxide dismutase, a cardinal enzyme in the cellular defense against oxidative damage. Second, rhCG signaling selectively limits activation of the apoptotic machinery in decidualizing HESCs by enhancing Bcl-2 expression whereas uhCG induces the expression of Fas ligand. Our results suggest that rhCG might be a preferable agent to protect the maternal decidua against oxidative damage in pregnancy, especially at the time of implantation and beyond. PMID- 21641642 TI - Tyrosine kinase inhibitors in the treatment of systemic mastocytosis. AB - Systemic mastocytosis (SM) is a heterogeneous disease, vast majority of these patients have a gain of function mutation in the gene encoding the tyrosine kinase KIT (KIT(D816V)). A small subset of SM patients with KIT(D816V) mutation require cytoreductive therapy. In these patients, tyrosine kinase inhibitors (TKIs) have been actively investigated over the last decade because of codon 816 KIT mutations causing constitutive activation of tyrosine kinase activity of the molecule. The main question has been whether the success story with imatinib in chronic myeloid leukemia (CML), another disease associated with a constitutively active tyrosine kinase, could be mimicked in mastocytosis. However, the results from various TKIs in SM with KIT(D816V) mutation have been disappointing to date. Only a few of the TKIs sufficiently block KIT(D816V) activity and have shown promising clinical results. The data from these studies indicate that, apart from KIT(D816V), other kinase targets and target pathways may play a role in disease evolution and progression, especially in patients with SM with an associated clonal hematological non-mast cell lineage disease (SM-AHNMD). Imatinib is effective in patients with increased mast cells and eosinophils associated with FIP1L1/PDGFRA+ (e.g., myeloid neoplasm with eosinophilia and rearrangement of PDGFRA) or rare patients with SM associated with KIT mutations outside of exon 17. This review will focus on the KIT receptor, KIT mutations, and the effects of the mutations in SM. The preclinical and clinical activities of FDA approved TKIs (for CML) as well as novel TKIs in SM will be evaluated. PMID- 21641643 TI - Predicting mixture toxicity of seven phenolic compounds with similar and dissimilar action mechanisms to Vibrio qinghaiensis sp.nov.Q67. AB - The predictions of mixture toxicity for chemicals are commonly based on two models: concentration addition (CA) and independent action (IA). Whether the CA and IA can predict mixture toxicity of phenolic compounds with similar and dissimilar action mechanisms was studied. The mixture toxicity was predicted on the basis of the concentration-response data of individual compounds. Test mixtures at different concentration ratios and concentration levels were designed using two methods. The results showed that the Weibull function fit well with the concentration-response data of all the components and their mixtures, with all relative coefficients (Rs) greater than 0.99 and root mean squared errors (RMSEs) less than 0.04. The predicted values from CA and IA models conformed to observed values of the mixtures. Therefore, it can be concluded that both CA and IA can predict reliable results for the mixture toxicity of the phenolic compounds with similar and dissimilar action mechanisms. PMID- 21641644 TI - Spontaneous cervical epidural hematoma mimicking stroke. PMID- 21641645 TI - Primary diffuse choroid plexus T-cell lymphoma: case report. AB - We present here a case of primary choroid plexus T-cell lymphoma with no evidence of immunodeficiency or immunological disease. As ventricular T-cell lymphoma is extremely rare, there is only limited information on the radiological findings of ventricular T-cell lymphoma. In this report, we also include some unusual MRI findings in this case that have never been described before. PMID- 21641646 TI - Bilateral Meckel's cave amyloidoma: a case report. AB - Primary solitary amyloidoma of Meckel's cave is rare, and a bilateral location is even more rare. To the best of our knowledge, only 12 cases in the literature have described such a primary lesion, including one case of bilateral involvement of Meckel's cave. We report here on the case of a 57-year-old woman presenting with pseudotumor masses involving both Meckel's caves and responsible for trigeminal neuropathy. The final diagnosis of amyloidoma was made on the basis of histological examination of surgical biopsy specimens. PMID- 21641647 TI - Inflammatory pseudotumors of the head and neck in pathology-proven cases. AB - The purpose of this case series is to characterize the CT and MRI features of pathology-proven inflammatory pseudotumors in the head and neck. Our search identified three orbital, one maxillary sinus, and one skull base inflammatory pseudotumor. All of the lesions demonstrated some degree of infiltrative features on imaging. On CT, all of the orbital inflammatory pseudotumors were of homogeneous soft tissue density. One of the orbital inflammatory pseudotumors demonstrated bone erosion and two others demonstrated stranding of the orbital fat. The maxillary sinus lesion initially appeared aggressive with bone erosion and orbital invasion. Calcifications were identified in the dural inflammatory pseudotumor. Among the lesions that were given contrast during CT or MRI. All exhibited some degree of enhancement. The two pseudotumors that underwent MRI were isointense on T1 and T2, with scattered areas of low signal. The orbital inflammatory pseudotumors underwent orbitotomy. However, the maxillary sinus and skull base lesions regressed with steroid therapy. Inflammatory pseudotumors of the head and neck regions typically manifest as enhancing soft tissue masses associated with infiltrative changes. Despite their sometimes-aggressive appearance, these lesions may respond well to steroid treatment. Imaging plays an important role in diagnosing and following inflammatory pseudotumors. PMID- 21641648 TI - AT1 receptor-mediated angiotensin II activation and chemotaxis of T lymphocytes. AB - Angiotensin II (Ang II), a central renin-angiotensin system (RAS) effector molecule, and its receptors, AT(1) and AT(2), have been shown to be involved in the inflammatory aspects of different diseases, however the cellular mechanisms underlying the regulation of immunity are not fully understood. In this work, using spleen-derived CD4(+) and CD8(+) T lymphocytes activated in vitro, we tested the influence of Ang II on different aspects of the T cell function, such as activation and adhesion/transmigration through endothelial basal membrane proteins. The addition of 10(-8)M Ang II did not change any of the parameters evaluated. However, 10(-6)M losartan, an AT(1) receptor antagonist: (i) reduced the percentage of CD25(+) and CD69(+) cells of both subsets; (ii) inhibited adhesion of these cells to fibronectin or laminin by 53% or 76%, respectively and (iii) significantly reduced transmigration through fibronectin or laminin by 57% or 43%, respectively. In addition, 10(-6)M captopril, an angiotensin-converting enzyme inhibitor had similar effects to Ang II, however its effects were reverted by exogenous Ang II (10(-8)M). None of these responses was modified by 10(-7)M PD123319, an AT(2) antagonist. These data reinforce the notion of endogenous production of Ang II by T cells, which is important for T cell activation, and adhesion/transmigration induced on interaction with basal membrane proteins, possibly involving AT(1) receptor activation. Moreover, AT(1) receptor expression is 10-fold higher in activated T lymphocytes compared with naive cells, but AT(2) receptor expression did not change after T cell receptor triggering. PMID- 21641649 TI - Changes in infants' affect related to the onset of independent locomotion. AB - Previous research suggests that after gaining several weeks of independent locomotor experience, infants may show both more negative and more positive affect toward parents. However, this prior work has been based largely on parent report, and no studies have used longitudinal or naturalistic methods to chart changes in infants' affective expressions as they gain locomotor ability. Fifteen infants were observed at home before, during, and after learning to crawl in two naturalistic contexts, free play and dyadic play. Expressions of negative affect during free play decreased after the onset of crawling, but there was no change in expressions of positive affect. At the same time, however, mothers reported an increase in both negative and positive reactivity. These results are discussed in terms of the contexts typically assessed during observations and the different sensitivities of mothers to infants' expressions of affect. Several lines of evidence point to a potential role for independent locomotion in the reorganization of affective expressions. PMID- 21641650 TI - Longitudinal study of PTSD, depression, and quality of life among adolescents after the Parnitha earthquake. AB - OBJECTIVE: To investigate the course of PTSD, depression, and current quality of life among adolescents 32-months after the 1999 Parnitha earthquake in Greece. METHODS: The follow-up was conducted among 511 adolescents originally evaluated at 3-months post-earthquake using the UCLA PTSD Reaction Index (PTSD-RI), Depression Self-Rating Scale (DSRS), and Quality of Life Questionnaire (QOLQ). RESULTS: Mean PTSD scores for the whole sample had subsided to mild levels; however, 8.8% were still experiencing moderate to severe levels of symptoms, and 13.6% met criteria for clinical depression. Frequency of experiencing reminders of the earthquake in the past month best explained the variance (15%) in PTSD severity, followed by depression at 3-months (8%). The QOLQ domain scores were negatively correlated with PTSD and depression. Depression at 3-months was the best predictor of QOLQ at 32-months, explaining 16% of the variance. LIMITATIONS: Self-report instruments were used; hence the responses may have been over- or under-estimated; also, the findings may not be generalizable to other ethnic groups. CONCLUSION: Ongoing screening is recommended after disaster to identify adolescents who continue to experience moderate to severe levels of PTSD and depressive symptoms. Specific interventions to reduce reactivity to earthquake related reminders should be a component of post-disaster recovery programs. A quality of life measure can provide important information in addition to traditional scales for monitoring the course of recovery among adolescents after disasters. PMID- 21641651 TI - Can effects of antidepressants in patients with mild depression be considered as clinically significant? AB - BACKGROUND: How to define clinical significance of antidepressants has become a matter of far-reaching clinical and regulatory consequences. A mean difference of at least 3 points on the Hamilton Depression Rating Scale (HAMD-17) between active treatment and placebo has been proposed as cut-off score for clinical significance in antidepressant trials. OBJECTIVE: We aimed to present arguments that this, and other commonly used related approaches to establish clinical significance are likely to be misleading and risky depriving patients with mild depression of efficient treatments. METHODS: These problems are exemplified with the data from a randomized placebo-controlled five-arm clinical trial with primary care patients with milder depressive syndromes (MIND-study). RESULTS AND CONCLUSIONS: Designs for studying clinical significance have to be distinguished from those assessing efficacy. Moreover, evaluation of the clinical significance of psychotherapy as a possible alternative to antidepressants faces the problem of how to define a valid control group where blinding of neither therapists nor patients is possible. PMID- 21641652 TI - The effects of postnatal maternal depression and anxiety on the processing of infant faces. AB - BACKGROUND: Postnatally depressed mothers have difficulties responding appropriately to their infants. The quality of the mother-child relationship depends on a mother's ability to respond to her infant's cues, which are largely non-verbal. Therefore, it is likely that difficulties in a mother's appraisal of her infants' facial expressions will affect the quality of mother-infant interaction. This study aimed to investigate the effects of postnatal depression and anxiety on the processing of infants' facial expressions. METHOD: A total of 89 mothers, 34 with Generalised Anxiety Disorder, 21 with Major Depressive Disorder, and 34 controls, completed a 'morphed infants' faces task when their children were between 10 and 18 months. RESULTS: Overall, mothers were more likely to identify happy faces accurately and at lower intensity than sad faces. Depressed compared to control participants, however, were less likely to accurately identify happy infant faces. Interestingly, mothers with GAD tended to identify happy faces at a lower intensity than controls. There were no differences between the groups in relation to sad faces. LIMITATIONS: Our sample was relatively small and further research is needed to investigate the links between mothers' perceptions of infant expressions and both maternal responsiveness and later measures of child development. CONCLUSION: Our findings have potential clinical implications as the difficulties in the processing of positive facial expressions in depression may lead to less maternal responsiveness to positive affect in the offspring and may diminish the quality of the mother-child interactions. Results for participants with GAD are consistent with the literature demonstrating that persons with GAD are intolerant of uncertainty and seek reassurance due to their worries. PMID- 21641653 TI - Psychoeducational methods for patients suffering from depression: the knowledge seeking instrument (KSI). AB - OBJECTIVE: To develop and psychometrically assess a short instrument that can be easily used in clinical practice to measure knowledge-seeking behavior in patients suffering from depression. METHOD: We developed the knowledge seeking instrument (KSI), a self-report scale of three items to assess the number of hours spent in knowledge seeking behavior such as reading written materials, surfing the internet, or watching audio-visual tools. Experts in mood disorders (n=12) participated in the formal validity assessment of the instrument, and the developed instrument was administered to outpatients who were attending psychiatry clinic (n=63). All patients also completed a multiple choice question instrument to measure knowledge of depression, a Likert self report questionnaire to assess attitudes towards depression and its treatment, and an adherence to antidepressants scale. RESULTS: In addition to the empirical evidence for validity, there was 68% agreement among experts that the items were highly relevant in measuring behavior of knowledge seeking, providing evidence for content validity. There were significant correlations (p<0.05) between knowledge of psychological and biological treatments of depression and knowledge seeking reading scores. The internal consistency reliability (Cronbach's alpha) was 0.67 for the instrument. CONCLUSION AND SIGNIFICANCE: The KSI takes 2 min to complete. There is evidence for reliability, content, and criterion based concurrent validities. The KSI can be utilized to assess knowledge seeking behavior in patients with depression. PMID- 21641654 TI - Orbitofrontal dysfunction related to depressive symptomatology in subjects with borderline personality disorder. AB - OBJECTIVE: In order to explore the relationships of fronto-limbic dysfunction with the clinical features of borderline personality disorder (BPD), the authors investigated brain electrophysiological activity in BPD patients following stimulation with emotionally arousing images. METHODS: Seventeen non-medicated patients with borderline personality disorder were studied with magneto encephalography. Regional cortical activities were obtained by minimum norm estimate (MNE) of steady-state visual evoked fields (ssVEFs). Linear regression models were conducted to explore clinical correlates of brain activity. RESULTS: Although no interaction group * picture category * brain region was found, a significant interaction group * brain region appeared for orbito-frontal cortex (OFC). BPD patients showed significantly reduced magnetocortical activity in left OFC across all picture categories (F = 26.4; p<.05; F = 31.4). Left OFC activity was inversely correlated with depression score in the BDI (r: -0.48, p < 0.05), with score in the Cornell Dysthymia rating scale (r: -0.52, p < 0.05) and with the number of criteria met for depressive personality disorder (r: -0.44, p < 0.05). Left orbitofrontal activity was also inversely correlated with the global score in the GAF (r-0.63, p < 0.01). No correlations were found between OFC activity and impulsivity or global severity of BPD symptoms. CONCLUSIONS: Abnormal functioning at orbitofrontal areas in BPD could be related to the presence of affective symptomatology and is associated with greater functional deterioration of patients. PMID- 21641655 TI - Highway to health; or How prenatal factors determine disease risks in the later life of the offspring. AB - Fetal development is largely dependent on the mother. However, pregnancy maintenance and consequently fetal development are highly vulnerable and sensitive to disruption, triggered by, for example, prenatal stress challenge. Such prenatal stress challenge modulates the maternal endocrine and immune responses during pregnancy e.g. by decreasing levels of progesterone. Prenatal stress also has negative repercussions for the child's health later in life. It has been reported that prenatal stress increases the risk of the child to develop chronic immune diseases such as allergies and asthma. We therefore propose that prenatal stress challenge - associated with a decrease in maternal progesterone - impairs fetal immune development (immune ontogeny). Such impaired immune ontogeny carries over into postnatal life, rendering the child more prone to developing chronic immune diseases. This purported association urgently requires a fresh evaluation in order to identify biomarkers and cascades of events. In the present review, we outline candidate biomarkers involved in fetal immune ontogeny, which may be targets of prenatal stress challenge and subsequently determine offspring disease risk. Identification of these stress-sensitive biomarkers may allow detection of pregnant women at risk to deliver chronic immune disease-prone offspring. The creation of therapeutic interventions designed to prevent negative consequences of prenatal stress would then be within reach. PMID- 21641656 TI - Effects and effectiveness of heparin in assisted reproduction. AB - Despite significant improvements in treatment protocols and laboratory techniques, the baby take-home rate after assisted reproduction (ART) is disappointing, e.g. 20% in Germany. It is therefore understandable that various approaches are undertaken to improve the pregnancy and delivery rates after IVF and ICSI. Among others, the application of heparin is suggested to avoid implantation failure - partially irrespective of maternal thrombophilia. The applications of heparin for various indications are presented in this manuscript and critically discussed, taking into account current studies and meta-analyses. Furthermore, possible effects of heparin on the embryo-maternal dialogue and indications for heparin to prevent and treat thromboembolic disease during ART are summarized. A systematic literature search of the PubMed/Medline database was performed assessing all articles published before November 2010. "IVF", "ICSI", "assisted reproduction", "heparin", "side effect", "coagulation", "thrombosis", "thrombophilia", "thrombophilic disorder", "implantation", "implantation failure" and "placenta" were used as keywords. The available studies on heparin in assisted reproduction are characterized by heterogeneous inclusion criteria and a lack of proven effectiveness in special constellations. In conclusion, the application of heparin to improve ART outcome rates is not justified. PMID- 21641657 TI - Innate immune cell recruitment in the fetus and neonate. AB - Recruitment of innate immune cells from the vasculature into infected tissue is a key event in primary host defense against invading pathogens. This highly regulated process requires a functional interplay of specialized adhesion molecules and involves a series of steps leading from rolling of leukocytes along the endothelium to firm adhesion and finally transmigration. In the developing fetus, innate immune functions are ontogenetically regulated and show increasing maturation throughout gestation. Developmental differences in the innate immune response leave the neonate and especially the premature newborn at high risk of severe infections. Understanding the ontogeny of immune functions in the fetus and newborn is therefore essential for the prevention and treatment of neonatal infections. In this review, an overview will be given of the developmental aspects of innate immune cell recruitment including a discussion of controversial findings and open questions. PMID- 21641658 TI - Disorders of implantation--are there diagnostic and therapeutic options? AB - Recent developments in reproductive medicine address oocyte morphology, sperm analysis and embryo selection. However, in a subgroup of infertile couples, it is the embryo implantation process that is disrupted. Diagnostic tools to identify patients at risk of implantation failure are limited and therapeutic options are far away from being established. In this review we focus on selected possible causes and treatments of failed implantation. Reproductive surgery allows a proper first step diagnosis and therapy of recurrent implantation failure (RIF). Possible anatomical malformations and associated diseases with treatment options are mentioned. Diagnostic procedures in patients often focus on defining gene polymorphisms (like hereditary thrombophilia and p53) and determinants of endometrial receptivity including endometrial gene expression profiles. Although significant differences in gene expression have been identified, the study populations are quite small, some of the data conflicting and clinical significance has yet to be proven. Implantation requires a close interaction between the fetal trophoblast and the maternal endometrium with natural killer cells (NK cells) playing a main part at the feto-maternal interface during early pregnancy. Therefore this review also focuses on NK cell receptor expression and new immunomodulatory treatment options like G-CSF in RIF patients. PMID- 21641659 TI - Thyroid hormone receptors and reproduction. AB - Thyroid disorders have a great impact on fertility in both sexes. Hyperthyroidism and hypothyroidism cause changes in sex hormone-binding globulin (SHBG), prolactin, gonadotropin-releasing hormone, and sex steroid serum levels. In females, thyroid hormones may also have a direct effect on oocytes, because it is known that specific binding sites for thyroxin are found on mouse and human oocytes. There is also an association between thyroid dysfunction in women and morbidity and outcome in pregnancy. In males, hyperthyroidism causes a reduction in sperm motility. The numbers of morphologically abnormal sperm are increased by hypothyroidism. When euthyroidism is restored, both abnormalities improve or normalize. In women, the alterations in fertility caused by thyroid disorders are more complex. Hyper- and hypothyroidism are the main thyroid diseases that have an adverse effect on female reproduction and cause menstrual disturbances--mainly hypomenorrhea and polymenorrhea in hyperthyroidism, and oligomenorrhea in hypothyroidism. In recent studies, it has become evident that it is not only changes in serum levels of SHBG and sex steroids that are responsible for these disorders, but also alterations in the metabolic pathway. Adequate levels of circulating thyroid hormones are of primary importance for normal reproductive function. This review presents an overview of the impact of thyroid disorders on reproduction. PMID- 21641660 TI - Immunological regulation of trophoblast invasion. AB - In investigating the immunological regulation of trophoblast invasion, emphasis is frequently placed on the constitution of the implantation site and its specific characteristics. However trophoblast cells are able of invading into not only the uterine tissue, but in the case of ectopic pregnancies also invade into other tissues, where they are actively involved in the creation of a beneficial local environment by expressing a range of membrane bound and soluble factors. The similarities and differences in trophoblast invasion in uterine and tubal pregnancies concerning trophoblast cells and maternal leukocytes and their interaction with each other are discussed in this review. PMID- 21641661 TI - Glycodelin-A as a paracrine regulator in early pregnancy. AB - Glycodelin-A (GdA) is a glycoprotein secreted from the endometrial glands and decidual glandular epithelium. Given its abundance and ubiquitous distribution in the first trimester uterus, GdA may be involved in early placental development via its modulatory effect on immune and trophoblast cells. GdA inhibits activation and proliferation, and induces apoptosis of T cells. By selectively inducing Th1 cell death, GdA may shift the Th1/Th2 ratio at the feto-maternal interface. This is also achieved indirectly through enhanced expression of Fas in the Th1 cells, thus making them vulnerable to cell death through Fas ligand expressed on trophoblast, endometrial, and activated T helper cells. GdA also promotes secretion of the Th2 cytokines IL-6 and IL-13 from NK cells, and induces immunological tolerance of dendritic cells and apoptosis of monocytes. Specific glycosylation is a prerequisite for the biological activities of GdA. Reduction in alpha2-6 sialylation of GdA, as in gestational diabetes, is associated with impairment of its T cell apoptosis-inducing activities. This review integrates recent studies on GdA and its role as a paracrine regulator in early pregnancy. PMID- 21641662 TI - Fostering new investigators in Reproductive Immunology. AB - During the 8th European Congress of Reproductive Immunology, November 2010 in Munich, Germany, the European Society of Reproductive Immunology provided the opportunity for young investigators to present their work. Short talks from students and post-doctoral trainees were scheduled immediately after the keynote speakers in each session. The Society presented two "Young Investigator Awards" in basic science as well as in clinical application, sponsored by Elsevier. Here we present a summary of the nominees in a single article. The nominees were asked to give a guided interview to provide an insight into their motivation and career aspirations for the future. We hope that the Young Investigator Award might be an ongoing tool to motivate and encourage young investigators to stay in the field of reproductive immunology and to continue their research on the feto-maternal interface. PMID- 21641664 TI - Heart rate variability, overnight urinary norepinephrine, and plasma cholesterol in apparently healthy human adults. AB - OBJECTIVE: The aim of this study was to assess the relationship between autonomic nervous system activity as indexed by measures of heart rate variability and overnight urinary norepinephrine, and plasma cholesterol levels in a large sample of working adults. METHODS: The study population comprised 611 apparently healthy employees of an airplane manufacturing plant in Southern Germany. Heart rate variability was calculated as beat-to-beat intervals over the course of one 24 hour weekday measured with an ambulatory ECG recorder. Overnight urine collection and blood samples were also obtained. RESULTS: We found an inverse association between indices of vagally-mediated heart rate variability and plasma levels of total cholesterol, low density lipoprotein (LDL), and the ratio of LDL to high density lipoprotein (HDL) that remained significant in multivariate models after controlling for relevant covariates including norepinephrine. Urinary norepinephrine was not significantly related to any measure of cholesterol in multivariate models. CONCLUSIONS: We report here for the first time, in a large sample of healthy human adults, evidence supporting the hypothesis of a clinically relevant inverse relationship between measures of plasma cholesterol and vagally-mediated heart rate variability after controlling for sympathetic nervous system activity. This suggests an important role for the vagal control of plasma cholesterol levels in cardiovascular disease. PMID- 21641663 TI - Comorbid anxiety disorders and baseline medication regimens predict clinical outcomes in individuals with co-occurring bipolar disorder and alcohol dependence: Results of a randomized controlled trial. AB - Despite the high prevalence and detrimental impact of alcoholism on bipolar patients, the diagnostic and treatment factors associated with better or worse clinical outcomes in alcohol-dependent patients with bipolar disorder are not well understood. The present study investigated the prospective impact of baseline psychiatric comorbidities and treatment regimens on clinical outcomes in bipolar alcoholics. Data were drawn from an 8-week randomized controlled clinical trial of acamprosate for individuals (n=30) with co-occurring bipolar disorder and alcohol dependence. Depressive and manic symptoms, and alcohol craving and consumption were monitored longitudinally using standardized instruments. Path analysis was used to estimate the prospective associations between patient characteristics and outcomes. More than 50% of patients were diagnosed with at least one anxiety (76.7%) or drug dependence disorder (60.0%). Comorbid anxiety disorders were prospectively associated with increased depressive symptoms and alcohol use. Participants were prescribed an average of 2.6 psychotropic medications at baseline. Antipsychotics and anticonvulsants were prospectively associated with increased alcohol use; anticonvulsants and benzodiazepines were associated with increased alcohol craving. Antidepressants were associated with increased depressive symptoms. Conversely, lithium was associated with decreased alcohol craving and depressive symptoms. The findings from the present study suggest areas for future research in this population. PMID- 21641665 TI - Effects of thoracic epidural blockade on mortality of patients with idiopathic dilated cardiomyopathy and heart failure. PMID- 21641666 TI - Arrhythmogenic potential of oxidative stress in atrial myocytes. PMID- 21641667 TI - Body-surface adjusted aortic reference diameters for improved identification of patients with thoracic aortic aneurysms: results from the population-based Heinz Nixdorf Recall study. AB - BACKGROUND: Early identification of patients at risk for thoracic aortic aneurysm (TAA) has the potential of improving prognosis. So far, however, "normal" aortic dimensions are not well defined, rendering identification of patients with enlarged aortas difficult. In the present study we aimed to (1) establish age- and gender-specific distribution of thoracic aortic diameters and (2) to determine the prevalence of asymptomatic TAA in a population-based European cohort. METHODS: Diameters of ascending thoracic aorta (ATA) and descending thoracic aorta (DTA) were measured from electron beam computed tomography (EBCT) scans of 4129 participants aged 45 to 75 years from the Heinz Nixdorf Recall study. Age- and gender-specific percentiles were calculated for body-surface adjusted aortic diameters. Multivariable linear regression was used to evaluate the association between aortic diameters and cardiovascular risk factors including age, gender and body-surface area (BSA). RESULTS: Aortic diameters were generally greater in the ATA than in the DTA, and were greater in men than in women (ATA: 3.71 +/- 0.4 cm vs. 3.45 +/- 0.4 cm, p<0.0001; DTA: 2.82 +/- 0.3 cm vs. 2.54 +/- 0.3 cm, p<0.0001). Age, male gender, blood pressure and body-surface area were independently associated with aortic diameters in both ATA and DTA. Based on our measurements age- and gender-specific percentiles for indexed ATA and DTA diameters were computed. Aneurysms >= 5 cm were found in 12 (0.34%) out of the total of 4129 subjects. CONCLUSION: Since BSA was independently associated with increasing aortic diameters, correction of aortic diameters for BSA may be more helpful in order to reliably identify patients at risk for aneurysm formation. Based on the normal distribution of body-surface adjusted thoracic aortic diameters displayed in age- and gender-specific percentiles we suggest a cut-off point for aneurismal aortic diameter at the 95th percentile. PMID- 21641668 TI - Coconut left atrium. PMID- 21641669 TI - Potential of wine-associated lactic acid bacteria to degrade biogenic amines. AB - Some lactic acid bacteria (LAB) isolated from fermented foods have been proven to degrade biogenic amines through the production of amine oxidase enzymes. Since little is known about this in relation to wine micro-organisms, this work examined the ability of LAB strains (n=85) isolated from wines and other related enological sources, as well as commercial malolactic starter cultures (n=3) and type strains (n=2), to degrade histamine, tyramine and putrescine. The biogenic amine-degrading ability of the strains was evaluated by RP-HPLC in culture media and wine malolactic fermentation laboratory experiments. Although at different extent, 25% of the LAB isolates were able to degrade histamine, 18% tyramine and 18% putrescine, whereas none of the commercial malolactic starter cultures or type strains were able to degrade any of the tested amines. The greatest biogenic amine-degrading ability was exhibited by 9 strains belonging to the Lactobacillus and Pediococcus groups, and most of them were able to simultaneously degrade at least two of the three studied biogenic amines. Further experiments with one of the strains that showed high biogenic amine-degrading ability (L. casei IFI-CA 52) revealed that cell-free extracts maintained this ability in comparison to the cell suspensions at pH 4.6, indicating that amine-degrading enzymes were effectively extracted from the cells and their action was optimal in the degradation of biogenic amines. In addition, it was confirmed that wine components such as ethanol (12%) and polyphenols (75 mg/L), and wine additives such as SO(2) (30 mg/L), reduced the histamine-degrading ability of L. casei IFI CA 52 at pH 4.6 by 80%, 85% and 11%, respectively, in cell suspensions, whereas the reduction was 91%, 67% and 50%, respectively, in cell-free extracts. In spite of this adverse influence of the wine matrix, our results proved the potential of wine-associated LAB as a promising strategy to reduce biogenic amines in wine. PMID- 21641670 TI - Detection of E. coli O157:H7 in raw ground beef by PathatrixTM immunomagnetic separation, real-time PCR and cultural methods. AB - Detection of Escherichia coli O157:H7 by conventional cultural methods can be difficult in food matrices with a high background flora such as raw ground beef. Raw ground beef samples, artificially contaminated separately with five strains of E. coli O157:H7 at low (~0.2 cfu/g) and high (~2 cfu/g) levels, were enriched by two enrichment protocols; buffered peptone water (BPW) at 37 degrees C for 5h and 24h and modified buffered peptone water with pyruvate (mBPWp) for 5h at 37 degrees C followed by adding selective agents and incubating at 42 degrees C to 24h. Detection of added E. coli O157:H7 by real-time PCR (RTiPCR) and recovery on isolation agars was performed before and after PATHATRIXTM immunomagnetic separation (IMS). RTiPCR detection and cultural recovery of inoculated E. coli O157:H7 after 5h enrichment were poor at 0.21-0.24 cfu/g. The addition of IMS after 5h enrichment did not improve RTiPCR detection but markedly improved recovery by culturing. By extending enrichment to 24h, RTiPCR detection improved to 76% for either enrichment protocol without IMS. When 24h enrichment was followed by IMS, RTiPCR detection was also further improved. Cultural recovery after 24h enrichment was 56% and 84% without IMS and 100% and 92% after IMS for BPW and mBPWp respectively. Extended enrichment to 24h followed by IMS was found to be sensitive and reliable for detection and cultural recovery from raw ground beef using either enrichment method. PMID- 21641671 TI - Transcription factor 7-like 2 (TCF7L2) gene polymorphism and complication/comorbidity profile in type 2 diabetes patients. AB - Transcription factor 7-like 2 gene (TCF7L2) has been associated with type 2 diabetes. We investigated the association of the rs7903146 SNP in this gene with clinical profile of type 2 diabetes (T2DM) patients. The study involved 980 patients with diabetic nephropathy (44%), diabetic retinopathy (42%), CVD (65%) and early onset of diabetes (45%) and 924 healthy controls. Subjects were genotyped for rs7903146 by PCR-RFLP. Genotype frequencies significantly differed between T2DM patients and controls (p<0.01, odds ratio (OR) for TT genotype 2.49 (95% confidence interval (CI) 1.84-3.39). An association was observed between rs7903146 and nephropathy (p<0.001), with 22% of TT homozygotes in this subgroup vs. 11% in patients without nephropathy (p=0.006, OR for TT 2.83, 95% CI 1.94 4.13). Association was stronger in patients with early onset of diabetes (34% of TT vs. 12% in the late onset, p<0.001). In DN group 71% of TT homozygotes had an early onset (OR 7.64, 95% CI 4.98-11.73 vs. controls). Our results confirm association of rs7903146 in the TCF7L2 gene with increased risk of type 2 diabetes. The T allele is strongly associated with nephropathy, especially in early onset of diabetes. PMID- 21641672 TI - EGFR inhibitors with concurrent thoracic radiation therapy for locally advanced non-small cell lung cancer. AB - Currently, a combination of chemotherapy and radiotherapy is the standard treatment approach for locally advanced non-small cell lung cancer (NSCLC). However, the clinical outcomes are still disappointing, with the 5-year survival rate being only approximately 20%. Further improvement in treatment outcome for patients with locally advanced NSCLC will require the development of more effective combined-modality therapies. Increasing attention has focused on the integration of targeted agents into current therapies. Many preclinical studies in this area have targeted the epidermal growth factor receptor (EGFR) signaling pathway to increase radiosensitivity. The in vitro rationale for targeting EGFR and concurrent ionizing radiation is well established, but to date, rare clinical data could provide proof-of-principle. In this review article, we briefly discuss pre-clinical data and the rationale and report all the different published clinical trials focusing on efficacy and toxicity in order to clarify and to summarize the present state-of-the-art of this particular combination in NSCLC. PMID- 21641673 TI - Empirical approaches to metacommunities: a review and comparison with theory. AB - Metacommunity theory has advanced understanding of how spatial dynamics and local interactions shape community structure and biodiversity. Here, we review empirical approaches to metacommunities, both observational and experimental, pertaining to how well they relate to and test theoretical metacommunity paradigms and how well they capture the realities of natural ecosystems. First, we show that the species-sorting and mass-effects paradigms are the most commonly tested and supported paradigms. Second, the dynamics observed can often be ascribed to two or more of the four non-exclusive paradigms. Third, empirical approaches relate only weakly to the concise assumptions and predictions made by the paradigms. Consequently, we suggest major avenues of improvement for empirical metacommunity approaches, including the integration across theoretical approaches and the incorporation of evolutionary and meta-ecosystem dynamics. We hope for metacommunity ecology to thereby bridge existing gaps between empirical and theoretical work, thus becoming a more powerful framework to understand dynamics across ecosystems. PMID- 21641674 TI - Multiple bud cultures of 'Barhee' date palm (Phoenix dactylifera) and physiological status of regenerated plants. AB - Adventitious bud clusters of date palm 'Barhee' were successfully established from juvenile leaves (<1cm) using reduced amounts of 2,4-D (0.2mgL(-1)) to limit the risk of somaclonal variation. An average of 8.4 adventitious buds per explant were obtained. Histological examination showed that the superficial cell layers of leaves had the highest caulogenic capacity. High sucrose concentration (70gL( 1)) was used for the conversion of initial buds to multiple bud clusters. The promoting effect of temporary immersion on shoot proliferation was found to be significant when compared to cultivation on solid media. Elongation of shoots was also better using a thin film of PGR-free liquid medium instead of a solid medium. Anatomical observations indicated that roots from vitroplants were potentially functional at various developmental stages. However, only 12-month old vitroplants were found to be physiologically able to control transpirational vapor loss. Additionally, the photochemical activity of photosystem II in these vitroplants was close to that measured in plants that were already acclimatized. As a result, 83.3% of regenerated plants were successfully acclimatized. No phenotypic variation was observed among more than 500 adventitious bud-derived plants. All regenerants survived after field transplantation. We found that the production of adventitious bud clusters in small bioreactors was able to provide an efficient micropropagation system for date palm cv. 'Barhee'. An in vitro hardening step was a prerequisite for the successful transfer of vitroplants in soil. PMID- 21641675 TI - Brachioradial pruritus as a result of cervical spine pathology: the results of a magnetic resonance tomography study. AB - BACKGROUND: Brachioradial pruritus (BRP) describes a rare form of itching occurring at the dorsolateral part of the forearms. Recent case reports suggest that BRP may be attributed to cervical lesions or spine neoplasms. OBJECTIVE: We sought to determine the incidence of cervical spine changes in BRP and to correlate the localization of spinal lesions with the dermatomal presence of pruritus. METHODS: Magnetic resonance tomography (MRT) of the cervical spinal cord, a chest x-ray, and a skin biopsy were performed in 41 patients (28 female, 13 male; 59.0 +/- 10.6 years) with BRP. Patients completed an itch questionnaire (NeuroDerm Questionnaire) that included a dermatome chart and the Northwick Park Neck Pain Questionnaire. RESULTS: The patients marked the locations C5 (90.2%) and C6 (100%) on the dermatome chart. All patients had detectable MRT changes. In 80.5% of the patients, stenosis of the intervertebral foramen or protrusions of the cervical disk led to nerve compression. The location of the nerve compression lesions correlated significantly with the dermatomal localization of the pruritus (Spearman correlation coefficient 0.893; P < .01). No spinal neoplasm was observed, and 19.5% of the patients had degenerative changes without significant correlation to the dermatomal localization of pruritus. LIMITATION: No healthy control group without pruritus was investigated. CONCLUSION: BRP may result from cervical nerve compression, and rarely, it may also stem from degenerative changes. Our findings suggest that even slight cervical changes detected on MRT may alter itch afferents and lead to BRP. Spinal cord tumors are rare and should be ruled out by a cervical spine MRT. PMID- 21641676 TI - Reappraisal of the provisional entity primary cutaneous CD4+ small/medium pleomorphic T-cell lymphoma: a series of 10 adult and pediatric patients and review of the literature. AB - BACKGROUND: Primary cutaneous CD4(+) small/medium-sized pleomorphic T-cell lymphoma (PCSM-TCL) was defined as a provisional entity in the 2005 World Health Organization-European Organization for Research and Treatment of Cancer classification of cutaneous lymphomas. A limited number of reports describe a generally indolent but heterogeneous condition. OBJECTIVE: We reviewed the concept of PCSM-TCL when applied to our experience and to the published literature. METHODS: We conducted a retrospective chart review and a PubMed search to identify all reported cases of PCSM-TCL from 2005 to 2010. RESULTS: Ten patients, including 4 of the youngest described, were identified in our institution, and their clinical and pathologic features were analyzed. All had a benign clinical course. Ten reports of patients with PCSM-TCL were reviewed and commonalities were found within an otherwise variable spectrum of clinical presentation, pathology, and biologic behavior. LIMITATIONS: This study was retrospective, follow-up was short term in some cases, and data were limited in a number of published reports. CONCLUSIONS: Our experience of 10 cases that met the diagnostic criteria of the provisional entity PCSM-TCL highlights its occurrence in children and further supports characterization of this condition as a clonal T cell lymphoproliferative disorder with indolent behavior. PMID- 21641677 TI - Clinical differentiation of acute cutaneous graft-versus-host disease from drug hypersensitivity reactions. AB - BACKGROUND: The early diagnosis of acute cutaneous graft-versus-host disease (GVHD) is challenging as there are no specific features. Drug hypersensitivity reaction (DHR) is the most frequently encountered condition to differentiate from GVHD. OBJECTIVES: We sought to investigate the clinical characteristics of acute cutaneous GVHD in comparison with DHR. METHODS: A retrospective chart review was conducted for allogeneic hemopoietic-cell transplantation recipients with maculopapular rashes who had undergone hemopoietic-cell transplantation less than 100 days earlier. We analyzed the clinical characteristics of 22 patients with acute cutaneous GVHD in comparison with 17 patients with DHR. RESULTS: Facial involvement was more frequent in the GVHD group (59%) than in the DHR group (24%). The difference was more significant when both the face and palms/soles were involved (36% in the GVHD group, no patients in the DHR group). Diarrhea was significantly more common in the GVHD group (73%) than in the DHR group (12%). The presence of both diarrhea and hyperbilirubinemia occurred only in the GVHD group (41%). In the GVHD group, the patients developed diarrhea and hyperbilirubinemia gradually with most patients exhibiting diarrhea and hyperbilirubinemia after 2 days. LIMITATIONS: A retrospective design, small number of patients, and single-center design are limitations. CONCLUSIONS: Facial involvement, and moreover involvement of the face and palms/soles, suggests the diagnosis of GVHD versus DHR. The presence of diarrhea, and diarrhea accompanied by hyperbilirubinemia is thought to favor the diagnosis of GVHD. Rashes with more than 2 to 3 days of duration and not accompanied by diarrhea and hyperbilirubinemia are less likely to be GVHD. PMID- 21641678 TI - Monotherapy with inhaled colistin for the treatment of patients with ventilator associated tracheobronchitis due to polymyxin-only-susceptible Gram-negative bacteria. PMID- 21641679 TI - Decline in incidence of Clostridium difficile infection after relocation to a new hospital building with single rooms. PMID- 21641680 TI - Do probiotics reduce the duration and symptoms of acute infectious diarrhea? PMID- 21641681 TI - Economic impact of use of chlorhexidine-impregnated sponge dressing for prevention of central line-associated infections in the United States. AB - BACKGROUND: The economic impact of adding chlorhexidine gluconate (CHG) impregnated sponge dressing to standard care (ie, chg-impregnated sponge dressing + skin preparation and transparent film dressing vs skin preparation and transparent film dressing) for the prevention of central-line infections was evaluated. METHODS: Clinical and economic data were obtained from peer-reviewed published studies to populate the decision model. The efficacy of reducing catheter-related bloodstream infection (CR-BSI) incidence with CHG-impregnated sponge dressing came from 2 recent randomized controlled trials. One-way and two way sensitivity analyses were performed on key clinical and economic parameters. RESULTS: Based on model calculations, a hypothetical 400-bed hospital inserting 3,078 central venous catheters (CVCs) per year is expected to avoid an average of 35 CR-BSIs, 145 local infections, and 281 intensive care unit days annually with the systematic use of CHG-impregnated sponge dressing. Potential hospital net cost savings (mainly because of reduced CR-BSIs with use of the dressing) would be $895,000 annually. Results were robust across a range of values in sensitivity analyses. CONCLUSION: CHG-impregnated sponge dressing is a cost-effective CR-BSI prevention treatment option for patients requiring CVCs. The importance of these results should be considered in the context of federal government and insurance company policies that no longer permit enhanced reimbursement for CR-BSI. PMID- 21641682 TI - [Fatal mucormycosis not suspected in the ICU]. PMID- 21641683 TI - [Current state of recognition of the title in Intensive Care Medicine in the United Kingdom]. PMID- 21641684 TI - [Cutaneous necrosis due to injection of interferon-alpha 2b in a patient with chronic hepatitis C]. PMID- 21641685 TI - [Chronic hepatic encephalopathy in a patient with primary biliary cirrhosis]. AB - Chronic or persistent hepatic encephalopathy is a complication that develops in 1% of patients with chronic liver disease. We report a new case of this complication in a patient with primary biliary cirrhosis. A 69-year-old woman with stage IV primary biliary cirrhosis presented with a 6-month history of progressive memory deficits, tremors and somewhat clumsy gait. Examination revealed sub-jaundiced skin tone, short-term memory deficits, fine distal bilateral tremor in the upper extremities and generalized hyperreflexia with spread of the reflexogenic zone. The hemogram showed mild pancytopenia, hypertransaminasemia, cholestatic pattern, lengthened thromboplastin time and hypocholinesterasemia. Wilson's disease was excluded and a cranial magnetic resonance imaging scan showed a bilateral hyperintense globus pallidus on T1 weighted sequences, which, together with the symptoms, were compatible with the diagnosis. Chronic liver diseases may cause chronic hepatic encephalopathy. Gastroenterologists should be familiar with this entity. PMID- 21641686 TI - [Post-infectious functional gastrointestinal disorders: from the acute episode to chronicity]. AB - Functional gastrointestinal disorders (FGID) form a major part of gastroenterology practice. Several studies have reported the development of post infectious irritable bowel syndrome (PI-IBS) after acute gastroenteritis (AGE). Non-gastrointestinal (GI) infections may increase the risk of developing IBS. There are also data showing that a GI infection may trigger functional dyspepsia (PI-FD). The possible development of PI-IBS or PI-FD depends on factors related to both the infection and the host. Microinflammation has been found in patients with post-infectious FGID. Studies performed in animal models show that infection and acute inflammation permanently change gastrointestinal motility and sensitivity. The role of AGE in the development of FGID is important not only because this entity provides an excellent natural model for pathogenic study but also because it provides an opportunity for preventive action. PMID- 21641687 TI - [Cold tuberculous abscess mimicking an incarcerated inguinal hernia]. PMID- 21641688 TI - [Concept maps as a tool for the diagnosis of rare diseases]. AB - Rare diseases are a real public health problem for hospitals and also for primary care. We describe some metaphor-based diagnosis procedures, such as: "When you hear hoof beats don't always think horses, sometimes they could be zebras", or that one about the antiquarian who recognised a museum masterpiece while walking in the Rastro (Madrid). The "lightning diagnoses" by Skoda are an important historic record. T. Greenhalgh has tried to cover the gap between evidence based medicine and the intuitive diagnosis. We point out some clinical epidemiology rules in order to improve their early detection by family practitioners and paediatricians. In our opinion, the training in the diagnosis of rare diseases has to be different for primary care level and for hospital doctors. Concept maps are useful for diagnosis in primary care clinics. PMID- 21641689 TI - [Distribution of primary care expenditure according to sex and age group: a retrospective analysis]. AB - OBJECTIVE: To study the primary care expenditure per person and the weight of different health resources within sex and age groups. DESIGN: Cost analysis. Retrospective descriptive study. SETTING: 14 urban primary care centers (assigned population: 313,000). PARTICIPANTS: All patients who visited during 2008 (227,235). STUDY PERIOD: January to December 2008. MAIN MEASUREMENTS: Age, sex, visits, laboratory, radiology and complementary tests and referrals to specialists were obtained from patient electronic files. Pharmacy expenditure was obtained from invoices sent to the national health system by pharmacists. Fixed/semi-fixed costs were distributed among visits and a mean cost/visit was obtained. Costs were assigned for laboratory (mean application cost), for radiology and complementary tests (rate per type of test) and for referrals (adjusted rate). Descriptive analysis of data (median, interquartile range and coefficient of variance). RESULTS: Median expenditure was 362 ? (321 ? for men and 396 ? for women). For the up to 2 year-old group it was 410 ?, for aged 15-44 203 ? and 75 and above 1,255 ?.). The up to 2 years old visits represented 81% of total cost while pharmacy was 8%, while for those aged 75 and above visits accounted for 21% of the total cost and pharmacy 63%. CONCLUSIONS: Expenditure in primary care is higher in women, although the greatest differences were observed with age. In older than 74 years the median expenditure was six-fold higher than that for 3-44 years old group. In pediatrics the main source of expenditure was visits, representing 80% of the total in up to 2 years old. From 45 years old, pharmacy was the main source of expenditure and in those older than 74 it represented over 60% of the total. PMID- 21641690 TI - [The majority of doctors believe that it is good to take periodic recertification examinations]. PMID- 21641691 TI - [Chronic ulcerative lesions in an immunocompetent male]. PMID- 21641692 TI - Design, synthesis, bioconversion, and pharmacokinetics evaluation of new ester prodrugs of olmesartan. AB - Synthesis of new ester prodrugs of olmesartan is described. Their in vitro stabilities in simulated gastric juice, rat plasma, and rat liver microsomes were tested. And the pharmacokinetic parameters for olmesartan after their oral administration were also estimated and compared with those in case of olmesartan medoxomil. Compounds 13 and 14 demonstrated high stability in simulated gastric juice and were rapidly metabolized to olmesartan in rat liver microsomes and rat plasma in vitro. In addition, C(max) and AUC(last) parameters were significantly increased in case of compounds 13 and 14 compared with olmesartan medoxomil. These results indicate that compounds 13 and 14 with cyclohexylcarboxyethyl and adamantylcarboxymethyl promoieties, respectively, are promising prodrugs of olmesartan with markedly increased oral bioavailability. PMID- 21641693 TI - Synthesis and cytotoxic activity of the heptaphylline and 7-methoxyheptaphylline series. AB - Nineteen carbazole alkaloids modified from heptaphylline (I) and 7 methoxyheptaphylline (II) isolated from Clausena harmandiana were synthesized. Among these derivatives, Ih and IIi showed cytotoxicity against the NCI-H187 cell line with IC(50) values of 0.02 and 0.66 MUM, respectively, which are about 138 and 4 fold stronger than the ellipticine standard. In addition, oxime Ih displayed cytotoxicity against KB cells with an IC(50) value of 0.17 MUM which is about 10 times stronger than the ellipticine. This compound demonstrated weak cytotoxicity against Vero cells (IC(50) = 66.01 MUM). The results show convincingly that Ih may be a promising lead for the development of cytotoxic agents. PMID- 21641694 TI - Synthesis, characterization and biological evaluation of some 16beta-azolyl-3beta amino-5alpha-androstane derivatives as potential anticancer agents. AB - A series of new 16beta-azolyl-3beta-amino-5alpha-androstane derivatives were synthesized and characterized. The new compounds were screened for their anticancer activity against the human cancer cell lines SW480, A549, HepG2, HeLa and SiHa in vitro using the MTT assay. The results of the in vitro study showed that a number of compounds have shown IC(50) values lower than 20 MUM against the five cancer cell lines. PMID- 21641695 TI - Design, synthesis and anti-tubercular evaluation of new 2-acylated and 2 alkylated amino-5-(4-(benzyloxy)phenyl)thiophene-3-carboxylic acid derivatives. Part 1. AB - A series of 2-acylated and 2-alkylated amino-5-(4-(benzyloxy)phenyl)thiophene-3 carboxylic acid derivatives were synthesized and evaluated for anti-tubercular activity. Among these compounds, 10d, 15, 12h and 12k inhibited Mycobacterium tuberculosis (Mtb) growth with MIC values between 1.9 and 7.7 MUM and low toxicity against VERO cells. The four compounds were also tested against multidrug-resistant tuberculosis (MDR-TB) and extensively drug-resistant tuberculosis (XDR-TB) clinical strains, which were found to show moderate activity. In addition, molecular docking simulation was performed to position compounds 10d, 15, 12h and 12k into mtFabH active site to predict the probable binding mode. These studies thus suggest that the designed 2-amino-5 phenylthiophene-3-carboxylic acid scaffold may serve as new promising template for further elaboration as anti-TB drugs. PMID- 21641696 TI - Combined SVM-based and docking-based virtual screening for retrieving novel inhibitors of c-Met. AB - Aberrant c-Met activation has been demonstrated to be implicated in tumorigenesis and anti-cancer drug resistance. Discovery of c-Met inhibitors has attracted much attention in recent years. In this study, a support vector machine (SVM) classification model that discriminates c-Met inhibitors and non-inhibitors was first developed. Evaluation through screening a test set indicates that combined SVM-based and docking-based virtual screening (SB/DB-VS) considerably increases hit rate and enrichment factor compared with the individual methods. Thus the combined SB/DB-VS approach was adopted to screen PubChem, Specs, and Enamine for c-Met inhibitors. 75 compounds were selected for in vitro assays. Eight compounds display a good inhibitory potency against c-Met. Five of them are found to have novel scaffolds, implying a good potential for further chemical modification. PMID- 21641697 TI - Design, synthesis and biological evaluation of brain-specific glucosyl thiamine disulfide prodrugs of naproxen. AB - Glucosyl derivates exhibited favorable distribution to the brain. However, bidirectional transport of glucose transporter 1 might decrease concentrations of the prodrugs in brain before the release of parent drugs. To overcome this defect, glucosyl thiamine disulfide prodrugs 1a-1c incorporating naproxen were designed and synthesized. Furthermore, prodrug 2 and 3 were also prepared as control. The favorable physicochemical properties of these prodrugs were verified by stability and metabolism studies. Results from the in vivo distribution study indicated that 1a-1c, and 1b in particular, significantly increased the level of naproxen in brain when compared to 2 and 3. The study suggested glucosyl thiamine disulfide was a promising carrier to enhance the brain bioavailability of central nervous system active drugs. PMID- 21641698 TI - Non-substituted N-heteroaromatic selenosemicarbazone metal complexes induce apoptosis in cancer cells via activation of mitochondrial pathway. AB - We previously published the synthesis, characterization and cytotoxic effect of the novel Zn(II), Ni(II), and Cd(II) complexes with 2-formylpyridine selenosemicarbazone. Here we further investigate the mechanism of their antiproliferative activity against several cancer and vascular endothelial cell lines and compared it to the activity of the ligand itself, corresponding salts and, as a referent compound, cisplatin. Investigated complexes induced apoptosis in a time- and dose-dependent manner as well as changes in a cell cycle distribution. Caspase-3 activation in HeLa cells, MDA-MB-361 and vascular endothelial cells EA.hy 926 cells by ligand alone, as well as Zn(II), Ni(II), and Cd(II) complexes was preceded by the activation of the p53 tumor-suppressor gene family protein p73. In addition to activation of p73, these compounds also trigger cytochrome C release by upregulation of Bax expression. The release of cytochrome C has been linked to loss of mitochondrial membrane potential. However, our data indicated that the increased phosphorylation of ERK could be also one of the mechanism involved in the Zn(II), and Cd(II) complexes- induction of apoptosis. Selenosemicarbazone complexes with Cd(II) and Ni(II), possess dual ability to induce apoptosis as well as necrosis, and might present an added advantage for inducing cell death in a diverse array of malignant cells. Taken together, our findings could indicate potential role of these complexes as activator of cross-talk between different signaling pathways that leads to cell death, and thus making the complex intriguing field for further scientific, and maybe clinical investigations. PMID- 21641699 TI - Design, synthesis and antimicrobial activity of chiral 2-(substituted-hydroxyl)-3 (benzo[d]oxazol-5-yl)propanoic acid derivatives. AB - Chiral 2-(substituted-hydroxyl)-3-(benzo[d]oxazol-5-yl)propanoic acid derivatives were synthesized and their antibacterial activities were evaluated against fungus, Gram-negative and Gram-positive bacteria. In general, these compounds showed in vitro activities against all screened Gram-negative and Gram-positive bacteria, but poor MIC values for fungus Candida albicans. Remarkably, the (S) configuration-substituted phenoxyl side chain on position 2 of propanoic acid exerted excellent antibacterial activity against all screened bacteria. Preliminary structure-activity studies revealed that the hydrophobic substitutes, para-tert-butyl (11r), para-phenyl (11s) and para-benzyloxy (11t) on the phenoxyl side chain displayed best activities against all Gram-negative and Gram-positive bacteria with MIC values between 1.56 and 6.25 MUg/mL. PMID- 21641700 TI - Synthesis and biological evaluation of 1-(4'-Indolyl and 6'-Quinolinyl) indoles as a new class of potent anticancer agents. AB - A novel series of the biheterocycles-based compounds with core structure distinguished from combretastatin A-4 (1) and colchicine (5) have been synthesized and evaluated as potent anti-mitotic agents. Compound 1-(4'-Indolyl and 6'-quinolinyl)-4,5,6-trimethoxyindoles 13 and 19 showed substantial anti proliferative activity against various human cancer cell lines, regardless to the tissue origin and the expression of multiple-drug resistance MDR1, with a mean IC(50) value of 38 and 24 nM respectively. Compound 13 (IC(50) = 1.7 MUM) also exhibited similar anti-tubulin activities to 1 (IC(50) = 1.8 MUM) and displayed strong binding property to the colchicine binding site on the microtubules. Computational modeling analysis revealed that the binding mechanism of compound 13 is similar to that of CA4. PMID- 21641701 TI - Student nurses' experiences of anxiety in the clinical setting. AB - It is known that some student nurses who experience anxiety during clinical experiences leave nursing education programs. If nurse educators can better understand the anxiety of student nurses during clinical experience, they will be able to develop educational interventions to minimize students' anxiety. Decreasing anxiety has a two-fold effect. First, when anxiety is decreased, learning may be increased. Second, decreasing anxiety may help alleviate the nursing shortage because more students complete their nursing education. This qualitative phenomenological study examines student nurses' perception of anxiety in the clinical setting. Situated cognition learning theory is the theoretical framework. The main method of data collection is unstructured face-to-face interviews with 7 student nurses. The data was analyzed using a thematic analysis. The themes are reported in the rich descriptive words of the subjects. Implications for practice are discussed. PMID- 21641702 TI - Perceived acculturative stress and sense of coherence in Chinese nursing students in Australia. AB - University schools of nursing in Australia are the main providers of nursing education programmes for international students. The large cohort of overseas students requires that universities, as education providers, understand their needs and the problems related to studying and living in Australia. An exploratory descriptive quantitative study was conducted to investigate 119 Chinese international undergraduate nursing students' acculturative stress and sense of coherence at an Australian university in Sydney. Two validated scales were used: Acculturative Stress Scale for International Students (ASSIS) and Sense of Coherence (SOC). Results indicated that overall Chinese nursing students had a moderate level of acculturative stress and sense of coherence. However, there was a significant difference in the level of acculturative stress among three groups of Chinese nursing students enrolled in the Bachelor of Nursing course, and SOC was negatively correlated with the level of acculturative stress. This study extends the knowledge of the experiences of Chinese nursing students studying and living in an English-speaking country. The study also highlights the need for universities to provide relevant support to overseas students to make their study journey smoother and more successful. PMID- 21641703 TI - Associations between the macronutrient composition of the evening meal and average daily sleep duration in early childhood. AB - BACKGROUND & AIMS: Short sleep duration in early childhood may increase the risk for chronic diseases in later life. Strategies to improve sleep duration are thus of interest. We investigated whether the nutritional composition of the evening meal is associated with children's sleep duration in the 2nd year of life. METHODS: Multivariable regression models included 594 participants of the DONALD Study with 3-days weighed dietary records and average daily sleep duration at age 1.5 and 2 years. RESULTS: Higher energy intakes with the evening meal were associated with a longer sleep duration (1 min/10 kcal, p = 0,01). With respect to absolute intakes, carbohydrates (0.8 min/g, p < 0.0001), especially from high GI foods (1.3 min/g, p < 0.01), and a higher GL (1.5 min/g GL, p < 0.01) were accompanied by longer sleeping time. A qualitative exchange of energy from protein by energy from carbohydrates from high GI foods was only associated with increased sleep duration in toddlers without (1.9 min/%E, p < 0.05), but not with nightly eating occasions (p > 0.4). CONCLUSIONS: The observed associations are in line with suggested sleep-improving effects of carbohydrates. Effect sizes suggest that the clinical relevance of nutritional composition for sleep duration is limited in healthy young toddlers. These observations and their possible importance for more vulnerable groups need to be confirmed in clinical trials. PMID- 21641704 TI - Experiences of pregnant women attending a lullaby programme in Limerick, Ireland: a qualitative study. AB - OBJECTIVE: to explore women's experience of singing lullabies in pregnancy and their understanding of possible benefits for themselves and their infants. DESIGN AND PARTICIPANTS: over four group sessions with musicians, six pregnant women, all expecting their first baby, learnt to sing three lullabies. Qualitative semi structured interviews were conducted approximately three months after the birth to capture the women's experiences. Participants were recruited at antenatal classes at a metropolitan maternity hospital in Ireland. FINDINGS: The women enjoyed participation in the lullaby project and all felt it benefited them and their babies. Themes uncovered included: (1) beyond words: music and the articulation of deep emotion; (2) a balm for the soul: the power and pleasure of beautiful music; and (3) music and the facilitation of infant development. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTISE: Singing lullabies in pregnancy had a soothing and calming effect and was a pleasant experience for participants. It was also an emotional experience and appeared to facilitate the expression of difficult emotion such as fear and anxiety. This effect is likely to have a significant and positive impact on well-being for pregnant women and as such warrants further exploration. PMID- 21641705 TI - Risk and reproductive decisions: British Pakistani couples' responses to genetic counselling. AB - How far does ethnicity/culture/religion mediate couples' responses to genetic risk? This paper examines the responses of 51 British Pakistani couples referred to a genetics clinic in southern England to counselling about recurrence risks for genetic problems in children. It is based on fieldwork conducted between 2000 and 2004 that combined participant observation of genetics consultations with interviews in respondents' homes. Interviews were conducted with 62 adults in connection with these 51 cases, of which 32 were followed through two or more clinical consultations and 12 through more than one pregnancy. Risk responses were categorized as: taking the risk; postponing; exploring risk management or dismissing the risk as irrelevant to current circumstances. Responses were cross referenced for associations with the severity of the condition, number of affected and unaffected children, availability of a prenatal test, age, gender, and migration history. I found that most couples were initially risk-takers who already had an unaffected child or children. Couples caring for living children with severe conditions were more likely to postpone. However, the risk responses of 15 couples changed over time, most towards and some away from risk management, reflecting changes in couples' appreciation of the severity of the condition and their subsequent reproductive experiences. The study highlights the diversity and dynamism of responses within one ethnic group and challenges stereotypes about cultural and religious responses to genetic risk. PMID- 21641706 TI - Relationship between distance to social gathering facilities and risk of trachoma for households in rural Tanzanian communities. AB - Few studies have examined the physical isolation of households with trachoma cases. Thus, in this study, we sought to examine the association between household isolation, as measured by distance to social gathering facilities, and risk of trachoma. We hypothesized that households located closer to such facilities would have a decreased risk of trachoma, due to a variety of social, economic, and cultural reasons. To test this hypothesis we conducted a case control study of 668 households (93 cases, 575 controls) in eight villages in Kongwa District, Tanzania, in 2007. Case households were defined as having a child aged 1-5 years with clinical signs of trachoma. Distance of household's place of residence to three main social gathering facilities - bars/cafes, religious establishments, and commercial/government center - was measured with a portable geographic positioning system. Multiple logistic regression analyses, which controlled for potential confounders and accounted for clustering, demonstrated increased risk of trachoma with increasing distance to social gathering facilities. Compared with distances of <=700 m, odds of trachoma were approximately two-fold higher for households living >1400 m from bars/cafes and from religious establishments, suggesting increased risk of trachoma for households at the fringes of communities. Targeting these isolated households with special programming along with dissemination through trusted social gathering facilities may improve effectiveness of current prevention efforts. PMID- 21641707 TI - Myocardial tuberculosis. PMID- 21641708 TI - [An intracardiac foreign body, and a few more]. PMID- 21641709 TI - [Assessment of dynamic coronary plaque changes and the clinical consequences in type-II diabetic patients: a serial intracoronary ultrasound study]. AB - INTRODUCTION AND OBJECTIVES: One of the aims of secondary prevention is to achieve plaque stabilization. This study sought to investigate the clinical consequences and predictive factors of the change in the type of plaque (CTP) as assessed by serial intracoronary ultrasound in type II diabetic patients with known coronary artery disease. METHODS: 237 segments (45 patients) from the DIABETES I, II, and III trials were included. Intracoronary ultrasound from motorized pullbacks (0.5mm/s) after index procedure and at 9-month angiographic follow-up was performed in the same coronary segment. Nontreated mild lesions (angiographic stenosis <25%) with >=0.5mm plaque thickening and >=5mm of length assessed by intracoronary ultrasound were included. As different types of plaques may be encountered throughout a given coronary lesion, each study lesion was divided into 3 segments for serial quantitative and qualitative analyses. Statistical adjustment by multiple lesion segments per patient (generalized estimating equations method) was performed. A CTP was defined as any qualitative change in plaque type at follow-up. At 1-year follow-up, major adverse cardiac events - death, myocardial infarction and target vessel revascularization) - were recorded. RESULTS: A CTP was observed in 48 lesions (20.2%) and occurred more frequently (52.1%) in mixed plaques. Independent predictors of CTP were glycated hemoglobin levels (odds ratio [OR] 1.2; 95% confidence interval [CI] 1.01-1.5; P=.04); glycoprotein IIb-IIIa inhibitors (OR 0.3; 95% CI 0.1-0.7; P=.004) and statin administration (OR 0.3; 95% CI 0.1-0.8; P=.02). At 1-year follow-up CTP was associated with an increase in major adverse cardiac events rate (CTP 20.8% vs non-CTP 13.8%, P=.008; hazard ratio=1.9, 95% CI 1.3-1.9, P=.01). CONCLUSIONS: Qualitative changes in mild stenosis documented by intracoronary ultrasound in type II diabetics are associated with suboptimal secondary prevention and may have clinical consequences. Full English text available from: www.revespcardiol.org. PMID- 21641710 TI - The future of cardiopulmonary resuscitation: what if a TEE probe could shock, sense and pace? PMID- 21641711 TI - Usefulness of routine laboratory parameters in the decision to treat refractory cardiac arrest with extracorporeal life support. AB - AIM: To evaluate the usefulness of routine laboratory parameters in the decision to treat refractory cardiac arrest patients with extracorporeal life support (ECLS). METHODS: Sixty-six adults with witnessed cardiac arrest of cardiac origin unrelated to poisoning or hypothermia undergoing cardiopulmonary resuscitation without return of spontaneous circulation (duration: 155 min [120-180], median, [25-75%-percentiles]) were included in a prospective cohort-study. ECLS was implemented under cardiac massage, using a centrifugal pump connected to a hollow fiber membrane-oxygenator, aiming to maintain ECLS flow >= 2.5 l/min and mean arterial pressure >= 60 mm Hg. RESULTS: Forty-seven of 66 patients died within 24 h from multiorgan failure and massive capillary leak. Of 19/66 patients who survived >= 24 h with stable circulatory conditions permitting neurological evaluation, four became conscious and were transferred for further cardiac assistance, while three became organ donors. Ultimately, one patient survived without neurologic sequelae after cardiac transplantation. Using multivariate analysis, only pre-cannulation peripheral venous oxygen saturation (SpvO2, 28% [15-52]) independently predicted inability to maintain targeted ECLS conditions >= 24 h (odds ratio for each 10%-decrease [95%-confidence interval]: 1.65 [1.21; 2.25], p=0.002). The area under the receiver-operating-characteristics curve was 0.78 [0.63; 0.93]. SpvO2 cut-off value of 33% was associated with a sensitivity of 0.68 [0.50; 0.83] and specificity of 0.81 [0.54; 0.96]. SpvO2 <= 8%, lactate concentration >= 21 mmol/l, fibrinogen <= 0.8 g/l, and prothrombin index <= 11% predicted premature ECLS discontinuation with a specificity of 1. CONCLUSION: SpvO2 is useful to predict the inability of maintaining refractory cardiac arrest victims on ECLS without detrimental capillary leak and multiorgan failure until neurological evaluation. PMID- 21641712 TI - A role for measurement of nasal IgE antibodies in diagnosis of Alternaria-induced rhinitis in children. AB - BACKGROUND: Rhinitis is a very common disease, frequently caused by sensitisation to inhalant allergens. Negative results from skin prick tests (SPT) and in vitro IgE tests generally lead to a diagnosis of non-allergic rhinitis. However, it is possible, as indicated by studies addressed with dust mites or pollens that the production of specific IgE occurs exclusively at nasal level. METHODS: We measured specific nasal IgE in children suffering from rhinitis in the periods when Alternaria spores were present in the air. All subjects underwent SPT with a standard panel of aeroallergens (Stallergenes, Milan, Italy) and, in the same session, to nasal IgE test (NT). Nasal provocation test (NPT) with Alternaria was used as reference. RESULTS: Fifty-six subjects were included in the study. Of them, 20 (37.5%) were positive to SPT and 45 (80.3%) were positive to NT. In particular, 11 subjects (19.6%) had a positive SPT and a negative NT; 36 (64.3%) had a negative SPT and a positive NT; and 9 (16.1%) were positive to both tests. Positivity of NT and NPT was observed in 36 patients (69.6%), while positivity of SPT and NPT was observed in 15 patients (26.8%). This difference was highly significant (p<0.0001). CONCLUSIONS: These findings suggest that sensitisation to Alternaria is frequently expressed by exclusive production of specific IgE in the nasal mucosa. Thus, measuring nasal IgE in children with rhinitis and negative SPT during the period of presence of Alternaria spores seems helpful to avoid a mistaken diagnosis of non-allergic rhinitis. PMID- 21641713 TI - Surgical management of infertility due to polycystic ovarian syndrome after failure of medical management. AB - OBJECTIVES: To evaluate surgical management and fertility and pregnancy outcome in women with polycystic ovarian syndrome (PCOS). STUDY DESIGN: Retrospective file review and follow-up of 74 consecutive women with PCOS resistant to citrate clomiphene (CC) who underwent ovarian drilling by fertiloscopy with bipolar energy, together with hysteroscopic surgery when indicated (Canadian TASK FORCE II-2). RESULTS: Of 77 files, only 3 women were lost to follow-up. Mean age was 30.2 years (SD 5.3) [29.0-31.4 CI 95%], and mean BMI 25.6kg/m(2) (SD 6.2) [24.2 27.0 CI 95%]. Pregnancy occurred after drilling in 47 cases (63%), spontaneously in 20 (27%), after ovarian stimulation in 5 (6.7%) and after in vitro fertilization in 22 (29.7%). Laparoscopic conversion was required in 5 cases (6.7%), due to failure to visualize the adnexa (n=3), or pelvic adhesions (n=1), or uterine hemorrhage (n=1). Hysteroscopy detected and simultaneously treated a uterine anomaly in 18 of 74 patients: uterine septum (n=10, 13%), T-shaped uterine cavity (n=3, 4%), endometrial polyp (n=2, 2.7%), endometrial hypertrophy (n=2, 2.7%), and synechiae (n=1, 1.3%). The mean overall delay to pregnancy was 11.1 months (SD 8.5) [8.7-13.5 CI 95%] and to spontaneous pregnancy, 7 months (SD 7.6) [3.7-10.3 CI 95%]. The mean follow-up was 23.4 months (SD 16.5) [18.1-28.7 CI 95%]. After multivariate analysis, the likelihood of pregnancy was significantly associated with previous ovarian stimulation by FSH (OR=2.28, 95% CI=1.08-4.83) and initial FSH level (OR=0.52, 95% CI=0.29-0.93). CONCLUSION: Ovarian drilling by hydrolaparoscopy is an effective treatment for CC-resistant PCOS. The high rate of associated uterine anomalies justifies simultaneous hysteroscopic surgery. PMID- 21641714 TI - Resource management as a key factor for sustainable urban planning. AB - Due to fast urbanization and increasing living standards, the environmental sustainability of our global society becomes more and more questionable. In this historical review we investigate the role of resources management (RM) and urban planning (UP) and propose ways for integration in sustainable development (SD). RM follows the principle of circular causation, and we reflect on to what extent RM has been an element for urban planning. Since the existence of the first settlements, a close relationship between RM, urbanization and technological development has been present. RM followed the demand for urban resources like water, energy, and food. In history, RM has been fostered by innovation and technology developments and has driven population growth and urbanization. Recent massive resource demand, especially in relation to energy and material flows, has altered natural ecosystems and has resulted in environmental degradation. UP has developed separately in response to different questions. UP followed the demand for improved living conditions, often associated to safety, good manufacturing and trading conditions and appropriate sanitation and waste management. In history UP has been a developing research area, especially since the industrial era and the related strong urbanization at the end of the 18th century. UP responded to new emerging problems in urban areas and became increasingly complex. Nowadays, UP has to address many objectives that are often conflicting, including, the urban sustainability. Our current urban un-sustainability is rooted in massive resource consumption and waste production beyond natural limits, and the absence of flows from waste to resources. Therefore, sustainable urban development requires integration of RM into UP. We propose new ways to this integration. PMID- 21641715 TI - Remediation of biodiesel wastewater by chemical- and electro-coagulation: a comparative study. AB - The remediation of biodiesel wastewater was carried out using chemical and electrochemical techniques. Initially the fatty acid methyl esters (FAME or biodiesel) and free fatty acids (FFA) were chemically removed from the wastewater using three types of mineral acids, H(2)SO(4), HNO(3) and HCl, at different pH values within the range of 1.0-8.0. Optimally, approximately 24.3 ml/l of FAME/FFA were removed from the wastewater when using H(2)SO(4) to set a final pH of 2.5 for 7 min. All pollutant levels were markedly reduced during this step. That is, approximately 38.94%, 76.32% and 99.36% of COD, BOD5 and oil & grease were respectively removed. The acidic aqueous phase left after the removal of the FAME/FFA phase was then treated by chemical- and electro-coagulation processes. The results demonstrated that both investigated treatment processes were effective for treating wastewater from a biodiesel production plant. The chemical coagulation provided a lower operating cost (1.11 USD/m(3)) compared with the electro-coagulation process (1.78 USD/m(3)). However, the latter process provided a better quality of wastewater compared with the former process, with the exception of the BOD levels. PMID- 21641716 TI - Radiologic evaluation of spontaneous or endoscopic third ventriculostomy: which technique is more useful? PMID- 21641717 TI - Hydrogen substitutes for the in situ generation of H2O2: an application in the Fenton reaction. AB - This study investigates the ability of formic acid, hydrazine and hydroxylamine to act as H(2) substitutes in conducting phenol degradation by Fenton reaction using in situ generated hydrogen peroxide. The processes were performed with semi heterogeneous (Pd/Al(2)O(3)+soluble Fe(2+)) and fully heterogeneous (FePd/Al(2)O(3)) catalytic systems under ambient conditions. In contrast to bulk H(2)O(2) production conditions, hydrazine is able to produce H(2)O(2)in situ followed by phenol degradation using Pd/Al(2)O(3)+Fe(2+) at pH 3 without the need for halide ions. However, a degree of mineralization exceeding 37% could not be achieved. The significant production of in situ H(2)O(2) at the inherent acidic pH of hydroxylammonium sulfate in the presence of Pd/Al(2)O(3)+Fe(2+) was also found to differ from the bulk production of H(2)O(2), in which no H(2)O(2) was detected. A remarkable degree of mineralization (ca. 65%) as well as fast phenol degradation during the reaction started at pH 7 over FePd/Al(2)O(3) may be an advantage of using hydroxylamine. On the other hand, using formic acid, H(2)O(2) was produced at a moderate rate, thereby achieving higher efficiency in the mineralization of phenol. Most importantly, the catalysts were more stable in the presence of formic acid than hydrazine or hydroxylamine. PMID- 21641718 TI - Removal of antimicrobials using advanced wastewater treatment. AB - Removal of numerous classes of pharmaceuticals from the municipal and industrial wastewater, using conventional wastewater treatment, is incomplete and several studies suggested that improvement of this situation would require the application of advanced treatment techniques. This is particularly important for the treatment of industrial effluents, released from pharmaceutical industries, which can contain rather high concentrations of antimicrobials. The aim of this work was to evaluate membrane bioreactors (MBRs), nanofiltration, reverse osmosis and ozonation, as well as their combinations, for the removal of antimicrobials from a synthetic wastewater which simulated highly contaminated industrial effluents. The study was performed using a mixture of four important classes of antimicrobials, including sulfonamides (SA), fluoroquinolones (FQ), macrolides (MAC) and trimethoprim (TMP). Performance of two different types of MBRs, Kubota and Zenon, was evaluated under different regimes regarding hydraulic retention time, total organic load and total nitrogen load. It was shown that elimination of SA in MBR treatment was very efficient, while the elimination of MAC, FQ, and TMP was incomplete. A mass balance of these contaminants in MBR suggested that microbial transformation represented the main mechanism, while only a small percentage was eliminated from the aqueous phase by adsorption onto sludge particles. Nanofiltration and reverse osmosis achieved high elimination rates however produced highly contaminated concentrate. High removal was achieved using ozonation, but further research is needed to characterize formed ozonation products. PMID- 21641719 TI - Formaldehyde removal by potted plant-soil systems. AB - Formaldehyde is a major indoor air pollutant. Formaldehyde removal from indoor air conduces to decrease the health risk for urban inhabitants. In this study, a dynamic chamber technique was employed to investigate formaldehyde removal by potted spider plant (Chlorphytum comosum), aloe (Aloe vera) and golden pothos (Epipremnum aureum) with potted soils. The results showed that the potted plant soil systems could remove formaldehyde from air in a long time. The spider plant soil system had the highest formaldehyde removal capacity compared with others. Higher metabolisms in plants and microorganisms in daytime may give a reasonable explanation for higher formaldehyde removal capacities for plant-soil systems in daytime. The order of formaldehyde removal capacity for the three plant species agreed well with the sequence of formaldehyde dehydrogenase activities from plant leaves. Formaldehyde removal by plant may be diffusion-limited rather than reaction-limited since the detached formaldehyde dehydrogenase activities from the leaves of the three plant species were higher than in vivo metabolic capacities. Formaldehyde in air can be largely absorbed and metabolized by the microorganisms in the potted soils indicating that further elevating formaldehyde removal capacity for plant-soil system will be realized by increasing exposed surface of potted soil. PMID- 21641720 TI - Immune response and performance of growing Santa Ines lambs to artificial Trichostrongylus colubriformis infections. AB - This study was carried out to evaluate the immune response and the impact of Trichostrongylus colubriformis infections on the performance of growing Santa Ines lambs. Thirty male lambs, 3-4 months of age, were maintained in individual pens and restrictively randomised by weight into 3 treatment groups: (1) infected group, artificially infected with 2500 T. colubriformis larvae, three times a week, for 13 weeks, and fed ad libitum; (2) Pair-Fed Group, non-infected and fed with the same amount of food consumed by the infected animal of the same class on the previous day; and (3) control group, non-infected and fed ad libitum. Refused feed was weighed daily to assess the food intake of each lamb. Animals were weighed weekly and blood and fecal samples were collected. At the end of the trial, all lambs were euthanized to determine worm burden and collect intestinal tissues and mucus samples for histological and immunological analysis. The infected group presented eosinophilia, increased number of inflammatory cells in the mucosa, in addition to an increased production of specific immunoglobulins against T. colubriformis, which partially prevented the establishment of infective larvae. As a consequence of parasitism, the infected lambs presented reduced serum albumin concentrations and demonstrated severe small intestine lesions, such as villous atrophy and epithelial erosion, which impaired the digestion and absorption of nutrients, causing a significant loss in performance. The infected group presented a 37% reduction in daily weight gain (107.26 +/- 10.8 g/day), when compared with the control group (171.07 +/- 7.15 g/day). The infected lambs also demonstrated the worst food conversion, i.e., each animal needed to consume on average 10.05 +/- 0.52 kg of food to gain 1 kg live weight. The voluntary hay intake depression in infected animals was small, and statistical difference (P<0.01) was seen only on two occasions (ninth and 12th weeks), in comparison with the control group. In conclusion, Santa Ines lambs infected with T. colubriformis demonstrated a reduction in performance, caused by the damages produced by the adult nematodes in intestinal mucosa, and also by the immunopathological changes associated with the infection. PMID- 21641721 TI - Evaluation of miltefosine for the treatment of dogs naturally infected with L. infantum (=L. chagasi) in Brazil. AB - Dogs naturally infected with Leishmania Infantum (=L. chagasi) were treated with miltefosine using different therapeutic regimens. The animals were evaluated for clinical evolution, biochemical parameters, parasite load (by real-time PCR), cytokine levels and humoral response. After treatment and during the following 24 months, there was progressive clinical improvement and complete recovery in 50% (7/14) of the treated animals. There was a decrease in the smear positivity of the bone marrow after treatment, and there was also a gradual and constant decrease in positive cultures at the end of the follow-up period. However, the PCR detection of parasite DNA remained positive. In general, all animals presented a significant increase in parasite load 6 months after treatment. The IFN-gamma levels in all the groups tended to increase during follow-up period, regardless of the miltefosine dose administered. The IL-4 and IL-10 levels of the animals tended to decrease during follow-up, except after 300 days when only IL 10 increased. The serum antibodies identified antigens that ranged from 116 kDa to less than 29 kDa in the Western blot assay. Furthermore, 300 days after treatment, qualitative and quantitative differences in the antigen profiles were observed. Antigens of 97 and 46 kDa were the most intensely recognized. Higher levels of antigen-specific Leishmania IgG were detected before and 300 days after treatment in all groups. Taking together, the improvement in the clinical symptoms was not followed by parasitological clearance, suggesting that treatment with miltefosine is not recommended, especially in endemic areas like Brazil, where children are the major victims and dogs are involved in the maintenance of the parasite cycle. PMID- 21641722 TI - The molecular characterization of an Eimeria and Cryptosporidium detected in Asian seabass (Lates calcarifer) cultured in Vietnam. AB - An intestinal Eimeria was previously reported as a significant pathogen of Asian seabass (Lates calcarifer) in nurseries in Vietnam. In the present study, both Eimeria and Cryptosporidium were detected by sequence analyses of fragments of the 18S rRNA gene amplified from these Vietnamese L. calcarifer tissues. Based on these analyses, the Eimeria from the Vietnamese L. calcarifer formed clades with the Eimeria detected in L. calcarifer tissues from Australia, but clustered separately from other known Eimeria and Goussia species. The Cryptosporidium detected in L. calcarifer from Vietnam clustered closest with C. parvum and C. hominis. In situ hybridization using DIG-labeled DNA probes generated from 18S PCR products on the Vietnamese L. calcarifer wax block tissues showed that this method could not be used to distinguish between Eimeria and Cryptosporidium, due to the conserved nature of the 18S locus. A previously published study on the morphology of parasite developmental stages and oocysts in the Vietnamese L. calcarifer tissues showed only an intestinal Eimeria infection. The Cryptosporidium could be present at very low levels undetectable by microscopy in intestines, or being ubiquitous, was a possible contaminant from feed or water. While molecular analysis is a very useful tool in the study of disease and identification of aetiological agents, this study reiterates the importance of demonstrating organisms in situ in tissues. PMID- 21641723 TI - Multitargeted receptor tyrosine kinase inhibition: an antiangiogenic strategy in non-small cell lung cancer. AB - In the United States, the leading cause of cancer-related deaths is lung cancer, of which more than 85% of cases are categorized as non-small cell lung cancer. The process of angiogenesis, which results in the formation of vasculature, is a complex and coordinated process that is required for cancer growth and metastasis. Pathways that promote angiogenesis have been targeted as a therapeutic approach in multiple types of cancer, including non-small cell lung cancer. Of these, the vascular endothelial growth factor pathway has been the most well studied, but more recently, the platelet-derived growth factor and fibroblast growth factor pathways have been identified as regulators of angiogenesis and potential mediators of resistance to vascular endothelial growth factor inhibition. Bevacizumab, a monoclonal antibody that binds to vascular endothelial growth factor, is currently the only antiangiogenic drug approved for the treatment of non-small cell lung cancer; however, several tyrosine kinase inhibitors that target vascular endothelial growth factor receptors as well as platelet-derived growth factor receptors and/or fibroblast growth factor receptors are being developed. This article reviews the role of the fibroblast growth factor and platelet-derived growth factor pathways in angiogenesis and provides a summary of dual (e.g., sorafenib, sunitinib) and triple (e.g., BIBF 1120, pazopanib) antiangiogenic tyrosine kinase inhibitors currently in development for the treatment of non-small cell lung cancer. PMID- 21641724 TI - Intestinal inflammation influences alpha-MSH reactive autoantibodies: relevance to food intake and body weight. AB - Autoantibodies reacting with alpha-melanocyte-stimulating hormone (alpha-MSH), an anorexigenic neuropeptide, are involved in regulation of feeding. In this work we studied if intestinal inflammation (mucositis) may influence alpha-MSH autoantibodies production relevant to food intake and body weight. Mucositis and anorexia were produced in Sprague-Dawley rats by methotrexate (MTX, 2.5mg/kg/day, for three days, subcutaneously). Plasma levels of total IgG and of alpha-MSH autoantibodies were measured during and after MTX-induced mucositis and were compared with pair-fed and ad libitum-fed controls. Effects of intraperitoneal injections of rabbit anti-alpha-MSH IgG (3 or 10 MUg/day/rat) on MTX-induced anorexia and on plasma alpha-MSH peptide concentration were separately studied. Here we show that in MTX rats, intestinal mucositis and anorexia were accompanied by decreased plasma levels of both total IgG and of alpha-MSH autoantibodies while refeeding was characterized by their elevated levels. In spite of similar food intake in MTX and pair-fed rats, recovery of body weight was delayed by at least 1 week in the MTX group. During refeeding and body weight deficit in MTX rats, alpha-MSH IgG autoantibody levels correlated negatively with food to water intake ratios. Injections of anti-alpha-MSH IgG induced a dose-dependent attenuation of food intake and body weight regain in MTX-treated rats accompanied by increased concentrations of alpha-MSH peptide which correlated positively with plasma levels of alpha-MSH autoantibodies. These data show that intestinal inflammation, independently from food restriction, affects general humoral immune response which may influence food intake and body weight control via modulation of alpha-MSH plasma concentration by alpha-MSH reactive autoantibodies. PMID- 21641726 TI - Oxytocin specifically enhances valence-dependent parasympathetic responses. AB - The evolutionarily highly conserved neuropeptide oxytocin seems to be involved in the regulation of complex forms of social behavior. It enhances the processing of positive social stimuli, reduces behavioral and neuroendocrine stress responses and modulates amygdala activity in humans. Moreover, it has been proposed that oxytocin dampens sympathetic nervous system activity. This hypothesis was tested in a double-blind, placebo-controlled study with 38 men either receiving 24 IU oxytocin intranasally or a placebo spray. While accomplishing an emotion classification task, electrodermal responses were measured as an index of sympathetic activity. Moreover, heart rate changes were recorded that are additionally mediated by the parasympathetic nervous system. Oxytocin enhanced differential heart rate responses to facial expressions as a function of the emotional valence, but had no effect on electrodermal activity or tonic measures of physiological arousal. These results indicate that oxytocin specifically modulates phasic activity of the parasympathetic nervous system which potentially reflects an increased motivational value of facial expressions following oxytocin treatment. Findings suggest that anxiolytic effects of oxytocin are not reflected in short-term sympathetic responses and may even be a consequence of rather than a prerequisite for improved social information processing. PMID- 21641727 TI - Sex steroids and connectivity in the human brain: a review of neuroimaging studies. AB - Our brain operates by the way of interconnected networks. Connections between brain regions have been extensively studied at a functional and structural level, and impaired connectivity has been postulated as an important pathophysiological mechanism underlying several neuropsychiatric disorders. Yet the neurobiological mechanisms contributing to the development of functional and structural brain connections remain to be poorly understood. Interestingly, animal research has convincingly shown that sex steroid hormones (estrogens, progesterone and testosterone) are critically involved in myelination, forming the basis of white matter connectivity in the central nervous system. To get insights, we reviewed studies into the relation between sex steroid hormones, white matter and functional connectivity in the human brain, measured with neuroimaging. Results suggest that sex hormones organize structural connections, and activate the brain areas they connect. These processes could underlie a better integration of structural and functional communication between brain regions with age. Specifically, ovarian hormones (estradiol and progesterone) may enhance both cortico-cortical and subcortico-cortical functional connectivity, whereas androgens (testosterone) may decrease subcortico-cortical functional connectivity but increase functional connectivity between subcortical brain areas. Therefore, when examining healthy brain development and aging or when investigating possible biological mechanisms of 'brain connectivity' diseases, the contribution of sex steroids should not be ignored. PMID- 21641725 TI - The neuroendocrinology of childhood trauma in personality disorder. AB - BACKGROUND: Childhood trauma has been associated with elevated central corticotropin releasing hormone (CRH) drive in adults meeting general DSM-IV criteria for personality disorder. It is not clear how this may be related to pituitary or adrenal responsiveness in personality disorder. It was hypothesized that high levels of childhood trauma would be associated with blunted cortisol and adrenocorticotropin releasing hormone (ACTH) response to the combined dexamethasone(DEX)/CRH test in adults meeting general DSM-IV criteria for personality disorder. METHOD: 24 healthy, medication free adults with personality disorder (N=16) and a group of healthy controls (N=8) underwent semi-structured diagnostic interviews and completed the Childhood Trauma Questionnaire (CTQ). Across two separate study sessions separated by at least a week, cerebrospinal fluid (CSF) was sampled by lumbar puncture for measurement of CRH concentration (N=17), and peripheral blood cortisol and ACTH levels were measured after challenge with DEX/CRH (N=24). RESULTS: As hypothesized, high CTQ score was associated with a blunted cortisol and ACTH response to DEX/CRH challenge. Indices of cortisol and ACTH response (peak level and area under the curve (AUC)) to DEX/CRH were in turn significantly negatively correlated with CSF CRH concentration. CONCLUSION: Childhood trauma in adults with personality disorder is associated with blunted cortisol and ACTH secretion following DEX/CRH challenge. These effects are independent of depression or posttraumatic stress disorder. Previous work would suggest that blunted pituitary-adrenal response is related to elevated central CRH drive. Corroborating this, CSF CRH levels were significantly and negatively correlated with peak level and AUC of both cortisol and ACTH. PMID- 21641728 TI - Chronic psychosocial stress induces reversible mitochondrial damage and corticotropin-releasing factor receptor type-1 upregulation in the rat intestine and IBS-like gut dysfunction. AB - The association between psychological and environmental stress with functional gastrointestinal disorders, especially irritable bowel syndrome (IBS), is well established. However, the underlying pathogenic mechanisms remain unknown. We aimed to probe chronic psychosocial stress as a primary inducer of intestinal dysfunction and investigate corticotropin-releasing factor (CRF) signaling and mitochondrial damage as key contributors to the stress-mediated effects. Wistar Kyoto rats were submitted to crowding stress (CS; 8 rats/cage) or sham-crowding stress (SC; 2 rats/cage) for up to 15 consecutive days. Hypothalamic-pituitary adrenal (HPA) axis activity was evaluated. Intestinal tissues were obtained 1h, 1, 7, or 30 days after stress exposure, to assess neutrophil infiltration, epithelial ion transport, mitochondrial function, and CRF receptors expression. Colonic response to CRF (10 MUg/kg i.p.) and hyperalgesia were evaluated after ending stress exposure. Chronic psychosocial stress activated HPA axis and induced reversible intestinal mucosal inflammation. Epithelial permeability and conductance were increased in CS rats, effect that lasted for up to 7 days after stress cessation. Visceral hypersensitivity persisted for up to 30 days post stress. Abnormal colonic response to exogenous CRF lasted for up to 7 days after stress. Mitochondrial activity was disturbed throughout the intestine, although mitochondrial response to CRF was preserved. Colonic expression of CRF receptor type-1 was increased in CS rats, and negatively correlated with body weight gain. In conclusion, chronic psychosocial stress triggers reversible inflammation, persistent epithelial dysfunction, and colonic hyperalgesia. These findings support crowding stress as a suitable animal model to unravel the complex pathophysiology underlying to common human intestinal stress-related disorders, such as IBS. PMID- 21641729 TI - Ovarian enzymatically active stromal cells can be a promoter of ovarian surface epithelial tumor. AB - Surface epithelial tumors (SETs) are the most common neoplasms of the ovary. They are traditionally thought derived from the ovarian surface or, as a recent hypothesis suggests, from various sources outside of ovary. Enzymatically active stromal cells (EASCs) are scattered in stroma of ovary, and characterized by their steroid-producing ability. With my observation of the increased EASCs near the epithelial cells of SETs, I hypothesize the epithelial cells of SETs can cause the increase of EASCs by converse adjacent stromal cells to EASCs; and EASCs, as a positive feedback, can prompt the proliferation of their neighbouring epithelial cells of SETs by secreting steroid hormone. PMID- 21641730 TI - Evolutionary medicine: bottle feeding, birth spacing, and autism. AB - To compensate for the high metabolic costs of lactation, the likelihood of re impregnation shortly after childbirth is normally reduced due to hormonal changes triggered by breast feeding during the postpartum period. Nowadays, however, bottle feeding as a substitute for breast feeding precludes such changes and leads to early postpartum reinstatement of fertility. We suggest that recent data showing the risk of autism goes up dramatically as the time between pregnancies goes down [1] may be a byproduct of bottle feeding. The decision to bottle feed your last child may unwittingly put your next child at risk of being autistic. PMID- 21641731 TI - Influence of different feeding systems on the growth performance and muscle development of Japanese Black steers. AB - This study investigated the growth performance and gene expression for muscle development between grass hay-fed (GH) and concentrate-fed (CT) steers. Daily gain and energy intake during the fattening period of the GH group were lower than those of the CT group. Analysis of C/EBPalpha, PPARgamma2, myosin heavy chain (MHC), and myostatin gene expressions was performed by real-time PCR. Expressions of C/EBPalpha and myostatin in semitendinosus and longissimus lumborum (LL) muscles were higher in the CT group than in the GH group at the end of fattening. In LL muscle, MHC expression at the end of fattening was greater in the GH group than in the CT group. These results suggest that regulation of adipogenesis and myogenesis by the expression of genes involved in muscle development might have occurred in the skeletal muscle of the GH group by the feeding of grass hay and/or because of the low energy intakes. PMID- 21641732 TI - Fluorescent HPLC for the detection of specific protein oxidation carbonyls - alpha-aminoadipic and gamma-glutamic semialdehydes - in meat systems. AB - Precise methodologies for the routine analysis of particular protein carbonyls are required in order to progress in this topic of increasing interest. The present paper originally describes the application of an improved method for the detection of alpha-aminoadipic and gamma-glutamic semialdehydes in a meat system by using a derivatization procedure with p-amino-benzoic acid (ABA) followed by fluorescent high-performance liquid chromatography (HPLC). The method development comprises i) the description of a simple HPLC program which allows the efficient separation of the ABA and the key standard compounds and ii) the optimization of the procedure for the preparation of a meat sample in order to maximize the fluorescent signal for both protein carbonyls. Furthermore, the suitability of this method is evaluated by applying the technique to porcine burger patties. The present procedure enables an accurate and relatively fast analysis of both semialdehydes in meat samples in which they could play an interesting role as reliable indicators of protein oxidation. PMID- 21641733 TI - Change in T2-fat saturation MRI correlates with outcome in cervical cancer patients. AB - PURPOSE: To compare pretreatment and midtreatment tumor intensity as measured by T2 fat-saturation (T2-FS) MRI and its association with treatment response in cervical cancer patients. METHODS AND MATERIALS: Weekly MRI scans were performed for brachytherapy planning on 23 consecutive patients with clinical Stage IB1 to IIIB cervical cancer treated with definitive chemoradiotherapy. These scans were performed on a 1.5-T clinical scanner using a specialized pelvic coil. Mean signal intensity from T2-FS imaging was calculated for each tumor voxel. Average tumor intensity and tumor volume were recorded pre- and midtreatment (at Weeks 0 and 4). All patients subsequently underwent routine follow-up, including periodic clinical examinations and fluorodeoxyglucose-positron emission tomography imaging. RESULTS: Mean follow-up for surviving patients was 14.5 months. Mean tumor volume at presentation was 49.6 cc, and mean midtreatment tumor volume was 16.0 cc. There was no correlation between initial tumor volume and pretreatment signal intensity (r=0.44), nor was there a correlation between pre- or midtreatment tumor volume with disease-free survival (p=0.18, p=0.08 respectively.) However, having at least a 30% drop in signal intensity from pretreatment to midtreatment was correlated with having disease resolution on posttreatment fluorodeoxyglucose-positron emission tomography imaging (p=0.05) and with disease-free survival (p=0.03.) Estimated disease-free survival at 22 months was 100% for patients with at least a 30% drop in tumor signal intensity compared with 33% for patients above this selected threshold (p=0.004). CONCLUSIONS: Longitudinal changes in T2-FS tumor intensity during chemoradiation correlated with disease-free survival in cervical cancer patients. Persistently high midtreatment tumor intensities correlated with a high risk of treatment failure, whereas large decreases in tumor intensity correlated with a favorable outcome. PMID- 21641735 TI - Serum intestinal fatty acid binding protein level for early diagnosis and prediction of severity of necrotizing enterocolitis. AB - BACKGROUND/AIM: Intestinal fatty acid binding protein (I-FABP) is found within cells at the tip of the intestinal villi, an area commonly injured in necrotizing enterocolitis (NEC). In this study, we aimed to investigate the value of serum I FABP in early diagnosis and predicting severity of NEC. METHODS: This prospective study was conducted between April 2009 and November 2009. The preterm infants with suspected NEC were included in the study. These infants were divided into two groups according to their final diagnoses; Group 1: Stage 1 NEC and Group 2: Stages 2-3 NEC (Group 2a: Stage 2 NEC, Group 2b: Stage 3 NEC). Healthy preterms were assigned to control group (Group 3). Serial blood samples were obtained from the patients at symptom onset, 24h and 72 h later. One blood sample was taken from the controls. Serum I-FABP levels were compared among the groups. RESULTS: Initial serum I-FABP concentrations were 324.0+/-165.8 pg/ml, 764.7+/-465.1 pg/ml, and 360.2+/-439.5 pg/ml in Group 1, Group 2a, and Group 2b, respectively, and all were significantly higher than those of the control group (76.9+/-115.9 pg/ml) (p<0.001). The serum I-FABP levels gradually decreased from the onset of the disease to 72nd hour in Group 1 and Group 2a (p=0.001). In Group 2b I-FABP concentrations slightly decreased at 24th hour of the disease and increased thereafter, but the difference was not significant (p=0.06). CONCLUSION: Serial measurements of I-FABP levels may be a useful marker for early diagnosis and prediction of disease severity in NEC. PMID- 21641734 TI - Low ponderal index is associated with decreased muscle strength and fatigue resistance in college-aged women. AB - Poor fetal growth is associated with decrements in muscle strength likely due to changes during myogenesis. We investigated the association of poor fetal growth with muscle strength, fatigue resistance, and the response to training in the isolated quadriceps femoris. Females (20.6 years) born to term but below the 10th percentile of ponderal index (PI)-for-gestational-age (LOWPI, n=14) were compared to controls (HIGHPI, n=14), before and after an 8-week training. Muscle strength was assessed as grip-strength and as the maximal isometric voluntary contraction (MVC) of the quadriceps femoris. Muscle fatigue was assessed during knee extension exercise. Body composition and the maximal oxygen consumption (VO(2)max) were also measured. Controlling for fat free mass (FFM), LOWPI versus HIGHPI women had ~11% lower grip-strength (P=0.023), 9-24% lower MVC values (P=0.042 pre-trained; P=0.020 post-trained), a higher rate of fatigue (pre- and post-training), and a diminished training response (P=0.016). Statistical control for FFM increased rather than decreased strength differences between PI groups. The PI was not associated with VO(2)max or measures of body composition. Strength and fatigue decrements strongly suggest that poor fetal growth affects the pathway of muscle force generation. This could be due to neuromotor and/or muscle morphologic changes during development e.g., fiber number, fiber type, etc. Muscle from LOWPI women may also be less responsive to training. Indirectly, results also implicate muscle as a potential mediator between poor fetal growth and adult chronic disease, given muscle's direct role in determining insulin resistance, type II diabetes, physical activity, and so forth. PMID- 21641736 TI - Sex differences in blood cell counts in the Lothian Birth Cohort 1921 between 79 and 87 years. AB - BACKGROUND: Haematological abnormalities are important to detect. In older adults there are few data that inform about normal ranges and the change in blood cell counts with ageing. These few data indicate that sex differences in blood cell counts persist beyond 70 years. OBJECTIVE: To determine normal ranges for blood cell counts and changes in counts and to examine the relationship between different blood cell counts for men and women. METHODS: The sample comprised Lothian Birth Cohort 1921 (LBC1921) participants seen at a mean of about 79 and 87 years of age. Full blood count was measured on each occasion. Also recorded were weight, smoking status, years of full-time education, and main occupation, serum B12, folate and creatinine. RESULTS: We examined blood cell counts in 495 community-resident people with mean age 79.1 (range 77.8-80.6) and repeated this in 157 of them at mean age 86.7 (range 85.7-87.4) years. Normal ranges were similar for men and women for all blood cell types at both ages, but men had persistently significantly higher mean red cell and monocyte counts, and women significantly higher platelet counts. Between 79 and 87 years, red cell, lymphocyte and eosinophil counts fell significantly, neutrophil and platelet counts increased, and monocyte count was stable. Worsening renal function was associated with a decrease in mean red cell count. CONCLUSION: Changes in blood cell counts in this cohort are consistent with persistent sex differences, age related changes in renal function (red cells), background inflammation (neutrophils and platelets) and immunosenescence (lymphocytes). PMID- 21641737 TI - The hot flush--a determinant for vascular effects of estrogen? PMID- 21641738 TI - Determination of venlafaxine in post-mortem whole blood by HS-SPME and GC-NPD. AB - Venlafaxine is a phenethylamine derivative widely prescribed for the treatment of depression which inhibits both serotonin and norepinephrine reuptake (SNRI). In treatment with antidepressants of patient with depression and other psychiatric disorders there is also increased risk of suicidal thought and behaviour. Several lethal intoxications involving venlafaxine usually among psychotic patients have been reported in the literature. Sample preparation is of the greatest significance for a successful toxicological analysis. The development of simple, effective and rapid extraction procedures of drugs from post-mortem biological samples is a challenge. Headspace-solid phase microextraction (HS-SPME) offers significant advantages such as simplicity, low cost, compatibility with analytical systems, automation and solvent-free extraction. The aim of our work was the optimization of a HS-SPME procedure for the determination of venlafaxine in post-mortem biological samples by gas chromatography (GC) with nitrogen phosphorous detection (NPD). Venlafaxine was extracted on 100 MUm Polydimethylsiloxone Coating-Red (PDMS) SPME fiber and determined by GC-NPD. Salt addition, extraction temperature, preheating and extraction time were optimized to enhance the recovery of the extraction from aqueous solution spiked with venlafaxine. Finally the developed procedure was applied to post-mortem biological samples of a fatally poisoned woman by venlafaxine. The drug was quantified in post-mortem blood gastric and oesophagus contents of the deceased woman. A simple and rapid procedure using HS-SPME was developed for sample preparation of venlafaxine in post-mortem biological samples prior to GC-NPD determination. Validation data was satisfactory, thus enabling application in the toxicological analysis of forensic samples. PMID- 21641739 TI - Detection of ethyl glucuronide in blood spotted on different surfaces. AB - This study aims to show that sensitive detection of ethyl glucuronide in dried blood spotted onto various surfaces after a period of 24h is feasible. At present, there is insufficient information how tightly ethyl glucuronide (EtG) binds to various materials and how easily it can be eluted. 4ml aliquots of blood samples obtained from seven volunteers after consumption of alcoholic beverages were applied to six different surfaces. After drying and a 24h-storage at 20+/-2 degrees C the samples were re-dissolved in water, and EtG was subsequently analyzed by a LC-MS Paul-type ion trap. A comparison was made between dried and corresponding fluid samples. EtG was detectable in all subjects' samples following consumption of alcohol. EtG was also detectable after a storage time of four weeks at 4 degrees C in whole blood that had been preserved with EDTA. EtG was detectable in all samples dried on different surfaces and its concentration remained relatively constant irrespective of the particular condition of the material. Detection of EtG in blood spots from the scene may indicate recent alcohol consumption in cases where collection of blood remained undone or could not be performed. PMID- 21641740 TI - [Giant carotid aneurysm revealing Behcet's disease: a new case]. PMID- 21641741 TI - Investigating static nonlinearities in neurovascular coupling. AB - Many statistical models of coupling between time changes of the band-limited power of neural signals and functional magnetic resonance imaging Blood Oxygenation Level Dependent (BOLD) signal time changes rely on linear convolution. The effect of nonlinear behaviors in single-trial relationships between neural signals and BOLD responses is rarely tested and included in models. Here we investigate whether using a static nonlinearity improves the prediction of single-trial BOLD responses from neural signals. A static nonlinearity is a nonlinear transformation of the convolution of neural responses which is implemented by the same nonlinear function for all time points. We evaluated this approach by applying it to simultaneous recordings of functional magnetic resonance imaging BOLD and band-limited neural signals (Local Field Potentials and Multi Unit Activity) from primary visual cortex of anaesthetized macaques. We found that using a simple polynomial static nonlinearity was sufficient to obtain highly significant improvements of the accuracy of single trial BOLD prediction over the accuracy obtained with linear convolution. This suggests that static nonlinearities may be a useful tool for a compact and accurate statistical description of neurovascular coupling. PMID- 21641742 TI - Wavelet-based edge correlation incorporated iterative reconstruction for undersampled MRI. AB - Undersampling k-space is an effective way to decrease acquisition time for MRI. However, aliasing artifacts introduced by undersampling may blur the edges of magnetic resonance images, which often contain important information for clinical diagnosis. Moreover, k-space data is often contaminated by the noise signals of unknown intensity. To better preserve the edge features while suppressing the aliasing artifacts and noises, we present a new wavelet-based algorithm for undersampled MRI reconstruction. The algorithm solves the image reconstruction as a standard optimization problem including a l(2) data fidelity term and l(1) sparsity regularization term. Rather than manually setting the regularization parameter for the l(1) term, which is directly related to the threshold, an automatic estimated threshold adaptive to noise intensity is introduced in our proposed algorithm. In addition, a prior matrix based on edge correlation in wavelet domain is incorporated into the regularization term. Compared with nonlinear conjugate gradient descent algorithm, iterative shrinkage/thresholding algorithm, fast iterative soft-thresholding algorithm and the iterative thresholding algorithm using exponentially decreasing threshold, the proposed algorithm yields reconstructions with better edge recovery and noise suppression. PMID- 21641743 TI - MR signal change in venous thrombus relates organizing process and thrombolytic response in rabbit. AB - Venous thrombus is subsequently organized and replaced by fibrous connective tissue. However, the sequential changes in venous thrombi are not reliably detected by current noninvasive diagnostic techniques. The purpose of this study is to reveal whether magnetic resonance (MR) can detect venous thrombus, define thrombus age and predict thrombolytic responses. Thrombus in the rabbit jugular vein was imaged with a 1.5-T MR system at 4 h and at 1, 2 and 4 weeks using three dimensional (3D) fast asymmetric spin echo T2-weighted (T2W) and 3D-gradient echo T1-weighted (T1W) sequences. The jugular veins were histologically assessed at each time point. Magnetic resonance imaging (MRI) was also performed in vivo before and 30 min after tissue plasminogen activator (t-PA) administration. The thrombi in MRI were comparable in size to histological sections. The signal intensity (SI) of thrombi at 4 h was heterogeneously high or low on T2W or T1W images, respectively. The SI of thrombi on T2W images decreased time-dependently, but increased on T1W images at 1 and 2 weeks. Morphological analysis showed time dependent decreases in erythrocyte, platelet and fibrin areas and time-dependent increases in smooth muscle cell, macrophage, collagen and iron areas. The t-PA administration significantly decreased thrombus volume at 4 h but not at 1, 2 and 4 weeks. Venous thrombosis can be reliably and noninvasively detected by MRI. Measurement of SI might support assessments of thrombus age and thrombolytic response. PMID- 21641744 TI - Simultaneous acquisition of phosphocreatine and inorganic phosphate images for Pi:PCr ratio mapping using a RARE sequence with chemically selective interleaving. AB - The ratio of inorganic phosphate to phosphocreatine (Pi:PCr) is a validated marker of mitochondrial function in human muscle. The magnetic resonance imaging rapid acquisition with relaxation enhancement (RARE) pulse sequence can acquire phosphorus-31 ((31)P) images with higher spatial and temporal resolution than traditional spectroscopic methods, which can then be used to create Pi:PCr ratio maps of muscle regions. While the (31)P RARE method produces images that reflect the content of the (31)P metabolites, it has been limited to producing an image of only one chemical shift in a scan. This increases the scan time required to acquire images of multiple chemical shifts as well as the likelihood of generating inaccurate Pi:PCr maps due to gross motion. This work is a preliminary study to demonstrate the feasibility of acquiring Pi and PCr images in a single scan by interleaving Pi and PCr chemical shift acquisitions using a chemically selective radiofrequency excitation pulse. The chemical selectivity of the excitation pulse evaluated and the Pi:PCr maps generated using the interleaved Pi and PCr acquisition method with the subject at rest and during exercise are compared to those generated using separate Pi and PCr acquisition scans. A paired t test indicated that the resulting Pi:PCr ratios for the exercised forearm muscle regions were not significantly different between the separate Pi and PCr acquisition method (3.18+/-1.53) (mean+/-standard deviation) and the interleaved acquisition method (3.41+/-1.66). This work demonstrates the feasibility of creating Pi:PCr ratio maps in human muscle with Pi and PCr images acquired simultaneously by interleaving between the Pi and PCr resonances in a single scan. PMID- 21641745 TI - Assessment of pulmonary artery stiffness using velocity-encoding magnetic resonance imaging: evaluation of techniques. AB - The loss of pulmonary artery (PA) compliance has significant pathophysiological effect on the right ventricle. Noninvasive and reliable assessment of PA wall stiffness would be an essential determiner of right heart load and a clinically useful factor to assess cardiovascular risk. Two MRI techniques have been proposed for assessing PA stiffness by measuring pulse wave velocity (PWV): transit time (TT) and flow area (QA). However, no data are available that compares the two techniques and evaluates their performance, especially over a wide range of PWV values or at 3.0-T, which is the purpose of the present study. Thirty-three patients with different heart conditions were imaged using optimized high-temporal resolution and high-spatial resolution velocity-encoding MRI sequences. Statistical analysis was conducted to study intermethod, interobserver and intraobserver variabilities. The PWV measurements using TT and QA techniques showed good agreement (P>0.1). The Bland-Altman analysis showed negligible differences between the two methods (mean+/-S.D.=0.11+/-0.35 m/s, correlation coefficient r=0.94). The repeated measurements showed low interobserver and intraobserver variabilities, although the S.D. of the differences was larger in the QA technique. The mean+/-S.D. of the TT/QA measurement differences were 0.05+/-0.2/0.0+/-0.36 m/s and 0.02+/-0.26/0.02+/-0.39 m/s for the interobserver and intraobserver differences, respectively. In conclusion, each technique has its own advantages and disadvantages. The two techniques result in similar measurements, although the QA method is more subjective due to its dependency on operator intervention. PMID- 21641746 TI - Pseudoaortic dissection. AB - A 72-year-old man was brought to our emergency department (ED) because of upper abdominal pain. Initial vitals at the triage station were significant only for high blood pressure. Computed tomography (CT) of the abdomen with contrast enhancement revealed an intimal flap over his descending aorta, the infrarenal part. The flap was found throughout the descending aorta until its bifurcation. Stanford type B dissection was initially suspected. However, a reconstructed CT discovered an interesting feature of the aorta. A high aortic bifurcation at the level of the second lumbar vertebrae, in conjunction with bilateral common iliac arteries, appeared in pair, masquerading the CT image as infrarenal aortic dissection. Actually, that was a variant in human anatomy. A reconstructed sagittal view of the CT scan is mandatory for a patient with abdomen pain to avoid misinterpretation of the radiographic image. PMID- 21641747 TI - The 12-lead electrocardiogram in patients with subarachnoid hemorrhage: early risk prognostication. AB - OBJECTIVE: The aim of this study was to investigate if the electrocardiographic (ECG) abnormalities assessed early in the emergency department (ED) are associated with the in-hospital mortality of the patients with spontaneous subarachnoid hemorrhage (SAH). METHODS: We studied prospectively a cohort of 222 adult patients with spontaneous SAH in an ED. A 12-lead ECG was performed for these patients in the ED. The patients were stratified into nonsurvivors and survivors based on the in-hospital mortality. The clinical characteristics, heart rate, corrected QT interval (QTc) and 7 predefined morphologic abnormalities were compared between these 2 groups of patients. RESULTS: Compared with the survivors (n=178), the nonsurvivors (n=44) had significantly slower heart rate (75+/-23 vs 83+/-16, P=.018) and more prolonged QTc (492+/-58 vs 458+/-40, P=.001). There were significantly higher frequency of occurrence of ECG morphologic abnormalities (66% vs 37%, P=.001) and nonspecific ST- or T-wave changes (NSSTTCs; 32% vs 12%, P=.015) in the nonsurvivors compared with those in the survivors. Multiple logistic regression model identified QTc (odds ratio, 1.0; 95% confidence interval, 1.0-1.0; P=.005) and NSSTTC (odds ratio, 3.3; 95% confidence interval, 1.0-10.7; P=.047) as the significant ECG variables associated with in-hospital mortality. CONCLUSIONS: The occurrence of NSSTTC and prolonged QTc assessed early in the ED are independently associated with the in hospital mortality in adult patients with spontaneous SAH. PMID- 21641748 TI - Metabolic acidosis in septic shock: is the Stewart theory the magic bullet? PMID- 21641749 TI - About ultrasound in pneumothorax. PMID- 21641750 TI - Maternal effects on ethanol teratogenesis in a cross between A/J and C57BL/6J mice. AB - Genetic factors influence adverse pregnancy outcome in both humans and animal models. Animal research reveals that both the maternal and fetal genetic profiles are important for determining the risk of physical birth defects and prenatal mortality. Using a reciprocal-cross breeding design, we investigated whether the mother's genes may be more important than fetal genes in determining risk for ethanol teratogenesis. Examination of possible synergistic genetic effects on ethanol teratogenesis was made possible by using two mouse strains known to be susceptible to specific malformations. Inbred A/J (A) and C57BL/6J (B6) mice were mated to produce four fetal genotype groups: the true-bred AcA and B6cB6 genotypes and the genetically identical AcB6 and B6cA genotypes (the F(1) genotype). Dams were administered either 5.8 g/kg ethanol or an isocaloric amount of maltose-dextrin on day 9 of pregnancy. Fetuses were removed by laparotomy on gestation day 18, weighed, and assessed for digit, vertebral, and kidney malformations. Digit malformations in the genetically identical F(1) ethanol exposed litters showed a pattern consistent with a maternal genetic effect (AcB6 [2%] and B6cA [30%]). In contrast, vertebral malformations were similar in all ethanol-exposed litters (AcA [26%], AcB6 [18%], B6cA [22%], and B6cB6 [33%]). The percentage of malformations did not differ between male and female fetuses, indicating sex-linked factors are not responsible for the maternal effect. Ethanol exposure decreased litter weights but did not affect litter mortality compared with maltose-exposed controls. This study supports the idea that genes influence malformation risk following in utero alcohol exposure. Specifically, maternal genes influence risk more than fetal genes for some teratogenic outcomes. No evidence supported synergistic genetic effects on ethanol teratogenesis. This research supports the conclusion that uterine environment contributes to determining risk of Fetal Alcohol Spectrum Disorder. PMID- 21641751 TI - Validation of a measurement device for instrumented quantification of anterior translation and rotational assessment of the knee. AB - PURPOSE: A test setup for clinical use in the awake and non-anesthetized patient measuring anteroposterior translation and rotation of the knee joint is the subject of validation. METHODS: A measuring device featuring fixation of the foot at 30 degrees of knee flexion with varus/valgus stress posts for the knee was developed. Tibial rotation (external/internal) was imposed with a torque of 2 Nm on the footrest with the ankle locked in dorsiflexion. Anterior translation of the tibia in relation to the femur was measured with a commercially available arthrometer. Measurements were performed in a neutral position, internal rotation, and external rotation. Intrarater and inter-rater reliability was validated in 10 healthy volunteers (Cronbach alpha). We examined 10 patients with isolated anterior cruciate ligament (ACL) rupture, as well as 10 patients with ACL rupture plus medial instability and 10 patients with additional lateral instability. Side-to-side differences were used for calculation. RESULTS: Comparison of healthy volunteers and subjects with isolated ACL rupture showed significant differences: internal rotation, 0.79 mm and 2.46 mm, respectively (P = .001); neutral position, 0.4 mm and 3.35 mm, respectively (P < .0001); and external rotation, 0.29 mm and 2.5 mm, respectively (P = .003). Significant differences (P = .008) were found between isolated ACL rupture and ACL rupture plus medial instability by use of the ratio of anterior translation in external rotation versus the neutral position. Inter-rater reliability was 0.948 in 10 healthy volunteers and 0.981 in 10 subjects with unilateral ACL rupture. Intrarater reliability in the volunteers was 0.829. CONCLUSIONS: By use of the developed measurement device, the "Laxitester" (ORTEMA Sport Protection, Markgroeningen, Germany), objective differentiation between isolated ACL rupture and ACL rupture plus additional medial instability is possible. Values for anterior translation are reliable and reproducible by different examiners and by the same examiner at different times. CLINICAL RELEVANCE: The Laxitester allows objectification of medial instability in combination with ACL injuries and provides a reference regarding the need for additional medial stabilization. Compared with the isolated measurement of anteroposterior translation, knee instability can be assessed in a more differentiated manner. PMID- 21641752 TI - Antimalarial activity of new acridinone derivatives. AB - Malaria is one of the major threats concerning world public health. Resistance to the current antimalarial drugs has led to searches for new antimalarial compounds. Acridinone derivatives have recently demonstrated to be active against malaria parasite. We focused our attention on synthesized new acridinone derivatives, some of them resulting with high antiviral and trypanocidal activity. In this study new derivatives of 10-alyl-, 10-(3-methyl-2-butenyl)- and 10-(1,2-propadienyl)-9(10H)-acridinone were evaluated for their antimalarial activity against Plasmodium falciparum. To assess the selectivity, cytotoxicity was assessed in parallel against human MRC-5 cells. Inhibition of beta-hematin formation was determined using a spectrophotometric assay. Mitochondrial bc(1) complexes were isolated from yeast and bovine heart cells to test acridinone inhibitory activity. This study resulted in the identification of three compounds with submicromolar efficacy against P. falciparum and without cytotoxic effects on human cellular line. One compound, IIa (1-fluoro-10-(3-methyl-2-butenyl) 9(10H)-acridinone), can be classified as hit for antimalarial drug development exhibiting IC(50) less than 0.2 MUg/mL with SI greater than 100. In molecular tests, no relevant inhibitory activity was obtained for our compounds. The mechanism of acridinones antimalarial action remains unclear. PMID- 21641753 TI - Purification of TAT-CC-HA protein under native condition, and its transduction analysis and biological effects on BCR-ABL positive cells. AB - BCR-ABL oncoprotein is the cause of chronic myeloid leukemia. The homologous oligomerization of BCR-ABL protein mediated by BCR coiled-coil (CC) domain plays an important role in ABL kinase activation. The HIV-1 TAT peptide has been used extensively for the introduction of proteins into cells. We recombinated a TAT-CC HA protein to interrupt the homologous oligomerization of BCR-ABL. The expression conditions for TAT-CC-HA were optimized. The TAT-CC-HA fusion protein was purified with Ni+-NTA resin. TAT-CC-HA fusion protein was added into the cultures of Ba/F3-p210, 32D-p210, K562, KU812, Ba/F3, 32D, and HL-60 cells. It was found that TAT-CC-HA could transduce into these cells. It was confirmed that TAT-CC-HA fusion protein was internalized by Ba/F3-p210, K562, and Ba/F3 cells and located in the cytoplasm observed by confocal laser scanning fluorescence microscope. The transduction of TAT-CC-HA fusion protein into K562 cells was in a dose-dependent and time-dependent manner. The result of coimmunoprecipitation assay indicated that TAT-CC-HA could interact with BCR-ABL in K562 cells. The effects of TAT-CC HA fusion protein on cell growth and apoptosis were detected by MTT test and flow cytometry. Our findings suggested that TAT-CC-HA fusion protein could specifically inhibit the growth of BCR-ABL positive cells, and specifically induce apoptosis of BCR-ABL positive cells, while not affect the growth and apoptosis of BCR-ABL negative cells. PMID- 21641754 TI - [One ulcer out of five not related to Helicobacter pylori or NSAID/aspirin]. PMID- 21641755 TI - [Hypertensive nephrosclerosis]. AB - Hypertensive nephrosclerosis is the leading cause of end stage renal disease (ESRD) in France, however, in prospective clinical trials of hypertension, ESRD accounts only for a small fraction of all events (incidence rate 0.2 to 0.4% by year). Hypertensive nephrosclerosis is characterized histologically by a series of vascular injury, none of which is truly specific and that can be observed also in obesity or normal aging. Hypertensive nephrosclerosis is mildly symptomatic, but the prognosis is never benign, due to cardiovascular and renal burden. This unspecific presentation may explain why the diagnosis of hypertensive nephrosclerosis is easily carried by excess, the main differential diagnoses are atherosclerotic ischemic renal disease, poorly symptomatic primitive nephropathies or the sequelae of unnoticed malignant hypertensive nephrosclerosis. The very high prevalence of hypertensive nephrosclerosis in populations from African ancestry has suggested a genetic predisposition. MYH9/APOL1 gene variants have recently been identified and are strongly associated with hypertensive nephrosclerosis, however the pathophysiological link between these variants and renal disease is still unclear. The treatment is mainly based on blocking the renin angiotensin system, especially when proteinuria is present. The target blood pressure is less firmly established, the latest data from the AASK study, however, do suggest a benefit on progression of lower values < 135/80 or even < 130/80 mmHg, especially in patients with proteinuria. PMID- 21641756 TI - What happens at the adjacent knee joint after total hip arthroplasty of Crowe type III and IV dysplastic hips? AB - We prospectively evaluated 30 hips of 22 patients who had normal knees with a mean age of 53.4 years (range, 38-72 years). In the early postoperative period, genu valgum deformity was observed in all knees. Of 22 patients, 17 complained of severe pain owing to strain in the medial collateral ligament and iliotibial tract. Postoperatively, the ipsilateral extremities of the patients were extended by a mean of 16.5 mm (8-25 mm). Q angles of the patients increased by a mean of 4.4 degrees +/- 2.5 degrees (P < .001). Although the Harris hip scores were improved (40.7-87.8 points), postoperative Lysholm-Gillquist knee scores were significantly reduced (92-76 points, P < .001). Reduction of displaced hips into the anatomical hip center and lengthening the extremity despite shortening procedure may lead to strain at the knee joint iatrogenically, particularly with the mechanical effect of tensor fascia lata, which results with changes in the knee biomechanics. PMID- 21641757 TI - Optimal irrigation and debridement of infected joint implants: an in vitro methicillin-resistant Staphylococcus aureus biofilm model. AB - Acute postoperative and acute, late hematogenous prosthetic joint infections have been treated with 1-stage irrigation and debridement with polyethylene exchange. Success rates, however, are highly variable. Reported studies demonstrate that detergents are effective at decreasing bacterial colony counts on orthopedic implants. Our hypothesis is that the combination of a detergent and an antiseptic would be more effective than using a detergent alone to decrease colony counts from a methicillin-resistant Staphylococcus aureus biofilm-coated titanium alloy disk simulating an orthopedic implant. In our study of various agents tested, chlorhexidine gluconate scrub (antiseptic and detergent) was the most effective at decreasing bacterial colony counts both prereincubation and postreincubation of the disks; pulse lavage and scrubbing were not more effective than pulse lavage alone. PMID- 21641758 TI - Pulmonary embolism prophylaxis in more than 30,000 total knee arthroplasty patients: is there a best choice? AB - Prophylaxis for pulmonary embolism (PE) prevention in total knee arthroplasty remains controversial. A joint registry evaluated venous thromboembolism prophylaxis and anesthesia impact on the incidence of PE, fatal PE, and death. Patients received mechanical prophylaxis alone or chemical with or without mechanical prophylaxis. The overall PE incidence was 0.45%; fatal PE, 0.01%; and death, 0.31%. The only significant difference in any outcome was the incidence of PE between Coumadin and mechanical prophylaxis alone. Variables associated with a higher incidence of PE were age, an American Society of Anesthesiologists score of 3 or higher, and the use of general anesthesia. Based on the findings, general anesthesia can be discouraged, and only Coumadin fared better than mechanical prophylaxis alone, whereas other forms of chemical prophylaxis revealed no significant differences. PMID- 21641759 TI - Revision total hip arthroplasty without bone graft of high-grade acetabular defects. AB - Mixed results have been reported with bulk and cancellous bone graft to fill defects during acetabular revision arthroplasty. Jumbo cups have been used to maximize host bone contact, and if adequate initial stability can be achieved, this approach may provide a superior long-term outcome. We retrospectively reviewed a consecutive series of 107 acetabular revisions performed using jumbo cups without bone graft. Bone defects were assessed using a validated radiographic classification system that yielded 64 hips with significant bone defects for inclusion. Mean change in American Academy of Orthopaedic Surgeons lower extremity core and pain scores and in Short Form-12 scores showed increases of 22.01, 37.52, and 17.08 points, respectively. Postoperative radiographs consistently demonstrated host bone ingrowth into the jumbo acetabular shells, except for 3 failures. Careful incremental reaming up to a size that optimizes host bone support and contact may eliminate the need for bone graft in most acetabular revision arthroplasties. PMID- 21641760 TI - Deformation of 1-piece metal acetabular components. AB - The success of metal bearings is dependent on several parameters. The effects of in vivo forces on the deformation of monoblock acetabular components have yet to be determined. The purpose of our study was to assess the amount of deformation with press-fit fixation of 1-piece metal acetabular components. Four manufacturers provided 1-piece metal acetabular components in each size (30 cups). Testing was conducted using a custom vise to simulate press-fit fixation, and measurements were performed with a Mitutoyo Test device (Aurora, Ill). Previously determined in vivo forces were used in the press-fit simulation. All components deformed under simulated in vivo applied loads. Component deformation ranged from 15 to 300 um. Larger cups with thinner walls to accommodate larger heads had the greatest deformation and often exceeded the range of reported clearances from the manufacturers (76-227 um). PMID- 21641761 TI - Gender is a significant factor for failure of metal-on-metal total hip arthroplasty. AB - Metal-on-metal (MoM) articulations offers low wear, larger head size, and increased stability. Reports of early failure are troubling and include failure of ingrowth and metal articulation problems such as metallosis, hypersensitivity, pseudotumor, and unexplained pain. This study investigates the survivorship of modern MoM articulations by gender. We reviewed 1589 primary MoM THA in 1363 patients, with minimum 2-year follow-up for 1212 hips. Follow-up averaged 60 months. There were 643 female patients and 719 male patients. The incidence of cup revision was significantly higher in women than in men (8.2% vs 2.7%; P = .0000), as was incidence of aseptic loosening (4.3% vs 1.1%; P = .0006), and failure for metal-bearing complications (2.2% vs 0.6%; P = .0126). There appear to be gender factors influencing the success of MoM THA, which may include hormonal, anatomic, or functional differences. PMID- 21641762 TI - Prosthetic joint infection caused by gram-negative organisms. AB - Traditionally, periprosthetic joint infections (PJIs) due to gram-negative organisms are considered more difficult to manage; however, little literature exists with regard to outcome of PJI caused by gram-negative organisms. We identified 277 patients with 282 culture-positive PJI receiving surgical treatment. Thirty-one joints were treated for gram-negative PJI. The gram negative group was then compared with the gram-positive and polymicrobial PJI. A single debridement and retention of prosthesis were successful in 70% (7/10) of isolated gram negative compared with 33.3% (13/39) of methicillin-sensitive gram positive, 48.9% (23/47) of methicillin-resistant gram positive, and 57.1% (4/7) of polymicrobial. Of those patients undergoing a planned 2-stage exchange, a successful reimplantation was performed in 52% (12/23) of gram-negative, 51% (52/103) of methicillin-resistant gram-positive, 69% (65/94) of methicillin sensitive gram-positive, and 0% (0/8) of polymicrobial PJI cases. These results indicate that PJI due to gram-negative pathogens, although less common, is difficult to treat and is associated with limited success. PMID- 21641763 TI - Approaching patients and family members who hope for a miracle. AB - A clinical problem may arise when caring for patients or their surrogates who prefer continued aggressive care based on the belief that a miracle will occur, despite a clinician's belief that further medical treatment is unlikely to have any meaningful benefit. An evidence-based approach is provided for the clinician by breaking this complex clinical problem into a series of more focused clinical questions and subsequently answering them through a critical appraisal of the existing medical literature. Belief in miracles is found to be common in the United States and is an important determinant of how decisions are made for those with advanced illness. There is a growing amount of evidence that suggests end-of life outcomes improve with the provision of spiritual support from medical teams, as well as with a proactive approach to medical decision making that values statements given by patients and family members. PMID- 21641764 TI - Assessing excessive reassurance seeking in the anxiety disorders. AB - Reassurance seeking has long been hypothesized to be a key factor in the maintenance of anxiety within contemporary cognitive-behavioral approaches to the conceptualization and treatment of anxiety disorders. However, empirical studies have lagged due to the absence of a reliable and valid measure of reassurance seeking. The present study sought to develop and examine the psychometric properties of a theoretically derived measure of reassurance seeking in treatment seeking participants with DSM-IV-TR (American Psychiatric Association, 2000) social phobia (n=116), generalized anxiety disorder (n=75), panic disorder with or without agoraphobia (n=50), and obsessive compulsive disorder (n=42). Participants (N=283) completed the Reassurance Seeking Scale (RSS), Depression Anxiety Stress Scale, Beck Anxiety Inventory, and Beck Depression Inventory-II. An exploratory factor analysis resulted in a coherent three factor solution reflecting the need to seek excessive reassurance regarding: (1) uncertainty about decisions, (2) attachment and the security of relationships, and (3) perceived general threat and anxiety. The RSS was found to possess good internal consistency and was moderately correlated with measures of anxiety, stress, and depression. The psychometric properties of the RSS appear promising for the promotion of programmatic research on reassurance seeking and its treatment in the anxiety disorders. PMID- 21641765 TI - Posttraumatic stress disorder symptoms, intimate partner violence perpetration, and the mediating role of shame processing bias. AB - Posttraumatic stress disorder (PTSD) may produce internal "threats to the self," which generate shame. Shame is theoretically and empirically linked to intimate partner violence (IPV) perpetration. We examined relations among PTSD, cognitive processing of shame-relevant information, and IPV perpetration. Forty-seven community participants completed an emotional Stroop task with shame-relevant and neutral words. Stimuli were presented supraliminally (i.e., until vocal response) and subliminally (i.e., below an individualized threshold of conscious awareness). Facilitated color-naming of shame-relevant words (thought to reflect congruence between shame and self-schemas) mediated the relation between PTSD severity and IPV perpetration frequency. Mediation results for subliminal stimuli suggest that biased processing of shame cues may occur preconsciously and potentially catalyze processes (i.e., expectations of rejection in ambiguous situations with one's partner; avoidance that minimizes discomfort and protects self-image) that lead to IPV perpetration. Psychotherapeutic approaches to PTSD and IPV should consider the role of facilitated processing of shame cues. PMID- 21641766 TI - Cognitive processing and acrophobia: validating the Heights Interpretation Questionnaire. AB - Three studies were conducted to examine the psychometric properties of a new scale: the Heights Interpretation Questionnaire (HIQ). This scale was designed to measure height fear-relevant interpretation bias to help assess the relationship between biased interpretations and acrophobia symptoms. Studies 1 (N=553) and 2 (N=308) established the scale's factor structure and convergent and discriminant validity among two large undergraduate samples. Study 3 (N=48) evaluated the predictive validity of the HIQ by examining how well the scale predicted subjective distress and avoidance on actual heights. Factor analysis resulted in four distinct factors, and results suggest that each of the factors, along with the full HIQ, have good reliability and validity. Additionally, the scale predicts subjective distress and avoidance on heights beyond self-reported acrophobia symptoms. Overall, the HIQ shows promise as a new tool to investigate cognitive processing biases in acrophobia. PMID- 21641767 TI - Donating blood: a meta-analytic review of self-reported motivators and deterrents. AB - Although research on blood donor motivation abounds, most studies have typically focused on small sets of variables, used different terminology to label equivalent constructs, and have not attempted to generalize findings beyond their individual settings. The current study sought to synthesize past findings into a unified taxonomy of blood donation drivers and deterrents and to estimate the prevalence of each factor across the worldwide population of donors and eligible nondonors. Primary studies were collected, and cross-validated categories of donation motivators and deterrents were developed. Proportions of first-time, repeat, lapsed, apheresis, and eligible nondonors endorsing each category were calculated. In terms of motivators, first-time and repeat donors most frequently cited convenience, prosocial motivation, and personal values; apheresis donors similarly cited the latter 2 motivators and money. Conversely, lapsed donors more often cited collection agency reputation, perceived need for donation, and marketing communication as motivators. In terms of deterrents, both donors and nondonors most frequently referred to low self-efficacy to donate, low involvement, inconvenience, absence of marketing communication, ineffective incentives, lack of knowledge about donating, negative service experiences, and fear. The integration of past findings has yielded a comprehensive taxonomy of factors influencing blood donation and has provided insight into the prevalence of each factor across multiple stages of donors' careers. Implications for collection agencies are discussed. PMID- 21641768 TI - Memory functioning and mental verbs acquisition in children with specific language impairment. AB - Memory and language operate in synergy. Recent literature stresses the importance of memory functioning in interpreting language deficits. Two groups of 50 children each, ages 8-12 were studied. The first group included children with specific language impairment, while the participants in the second group were typically developing children. The two groups, which were matched on age, nonverbal intelligence and varied significantly in verbal ability were examined, using a test battery of four memory functioning (phonological, working and long term memory) and five mental verb measures. The statistical analyses indicated that the two groups differed significantly in all language and memory measures; a logistic regression analysis revealed that within each main group existed nested subgroups of different developmental patterns with working and long-term memory measures as the most robust discriminate markers of classification. Language impaired children had more difficulties in the acquisition of mental verbs because they are less able to process and store phonological information in working memory and long-term lexicon. PMID- 21641769 TI - Family impact in intellectual disability, severe mental health disorders and mental health disorders in ID. A comparison. AB - Family impact (or family burden) is a concept born in the field of mental health that has successfully been exported to the ambit of intellectual disability (ID). However, differences in family impact associated with severe mental health disorders (schizophrenia), to ID or to mental health problems in ID should be expected. Seventy-two adults with intellectual disability clients of the Carmen Pardo-Valcarce Foundation's sheltered workshops and vocational employment programmes in Madrid (Spain), 203 adults diagnosed with schizophrenia from four Spanish Community Mental Health Services (Barcelona, Madrid, Granada and Navarra) and 90 adults with mental health problems in ID (MH-ID) from the Parc Sanitari Sant Joan de Deu Health Care Site in Sant Boi de Llobregat, Barcelona (Spain) were asked to participate in the present study along with their main caregivers. Family impact experienced by caregivers was assessed with the ECFOS-II/SOFBI-II scale (Entrevista de Carga Familiar Objetiva y Subjetiva/Objective and Subjective Family Burden Interview). In global terms, results showed that the higher family impact was found between caregivers to people with MH-ID. The interaction of both conditions (ID and mental health problems) results in a higher degree of burden on families than when both conditions are presented separately. There was also an impact in caregivers to people with schizophrenia, this impact being higher than the one detected in caregivers to people with intellectual disability. Needs of caregivers to people with disability should be addressed specifically in order to effectively support families. PMID- 21641770 TI - Control of angular momentum during walking in children with cerebral palsy. AB - Children with hemiparetic Cerebral Palsy (CP) walk with marked asymmetries. For instance, we have recently shown that they have less arm swing on the affected side, and more arm swing at the unaffected side. Such an increase in arm swing at the unaffected side may be aimed at controlling total body angular momentum about the vertical axis, although it was never investigated in this respect. In the current study, we thus investigated if participants with hemiparetic CP control angular momentum by compensatory movements of the unaffected arm. We measured gait kinematics of 11 CP children, and 24 age matched typically developing (TD) children, walking at both self-selected and fast walking speeds, and calculated angular momenta. We found that children with hemiparetic CP did not have a reduced angular momentum of the affected arm. However, they showed substantial increases in angular momentum generated by the legs, which were compensated by increased angular momentum of the unaffected arm. As a result, there were no differences in total body angular momentum between TD and CP children. Moreover, walking speed had no effect on total body angular momentum in both groups. These findings support the idea that angular momentum during walking is a controlled variable, even in children with hemiplegic CP. PMID- 21641771 TI - Re: "American Society of Echocardiography recommendations for quality echocardiography laboratory operations". PMID- 21641772 TI - Characterization of diastolic dysfunction in twin-twin transfusion syndrome: association between Doppler findings and ventricular hypertrophy. AB - BACKGROUND: Twin-twin transfusion syndrome (TTTS) complicates 10% to 15% of monochorionic twin pregnancies. Cardiovascular changes of variable severity, such as ventricular hypertrophy, atrioventricular valve regurgitation, and systolic dysfunction, occur predominantly in recipient twins (RTs). It was the purpose of this study to perform a detailed assessment of ventricular geometry and diastolic function between controls, donor twins (DTs), and RTs. METHODS: In this prospective, case-control study, two-dimensional, pulsed-wave, and Doppler tissue imaging were used to evaluate biventricular geometry and diastolic function in controls, DTs, and RTs. RTs were divided into two groups, severe and mild, on the basis of evidence of high central venous pressure. Specific variables evaluated included relative wall thickness, mitral valve and tricuspid valve E/A velocities, diastolic filling time corrected for heart rate, isovolumic relaxation time, and early (E') and late (A') diastolic myocardial velocities. RESULTS: A total of 120 fetuses (39 TTTS twin pairs and 42 controls) were compared. Increases in relative wall thickness and isovolumic relaxation time were seen in the mild group. In the severe group, further increases in relative wall thickness and isovolumic relaxation time as well as decreased diastolic filling time corrected for heart rate were accompanied by the appearance of a monophasic Doppler inflow profile and elevations in the E/E' ratio, consistent with elevated ventricular filling pressures. CONCLUSIONS: Concentric hypertrophy is observed in RTs affected by TTTS and is associated with impaired ventricular relaxation and shortened filling time. In severe cases, further decreases in diastolic filling time and Doppler signs of elevated ventricular filling pressures are present. PMID- 21641773 TI - Numbering questionnaires had no impact on the response rate and only a slight influence on the response content of a patient safety culture survey: a randomized trial. AB - OBJECTIVES: In self-completed surveys, anonymous questionnaires are sometimes numbered so as to avoid sending reminders to initial nonrespondents. This number may be perceived as a threat to confidentiality by some respondents, which may reduce the response rate, or cause social desirability bias. In this study, we evaluated whether using nonnumbered vs. numbered questionnaires influenced the response rate and the response content. STUDY DESIGN AND SETTING: During a patient safety culture survey, we randomized participants into two groups: one received an anonymous nonnumbered questionnaire and the other a numbered questionnaire. We compared the survey response rates and distributions of the responses for the 42-questionnaire items across the two groups. RESULTS: Response rates were similar in the two groups (nonnumbered, 75.2%; numbered, 72.8%; difference, 2.4%; P=0.28). Five of the 42 questions had statistically significant differences in distributions, but these differences were small. Unexpectedly, in all five instances, the patient safety culture ratings were more favorable in the nonnumbered group. CONCLUSION: Numbering of mailed questionnaires had no impact on the response rate. Numbering influenced significantly the response content of several items, but these differences were small and ran against the hypothesis of social desirability bias. PMID- 21641774 TI - Tea catechins prevent contractile dysfunction in unloaded murine soleus muscle: a pilot study. AB - OBJECTIVE: Extended periods of muscle disuse, physical inactivity, immobilization, and bedrest result in a loss of muscle mass and a decrease in muscle force, which are accompanied by an increase in oxidative stress. We investigated the effects of the intake of green tea catechins on unloading induced muscle dysfunction in tail-suspended mice. METHODS: Ten-week-old male BALB/c mice were fed a purified control diet or a diet containing 0.5% tea catechins for 14 d. Thereafter, the mice were subjected to continuous tail suspension for 10 d. On the final day, muscle mass, contractile force production, antioxidant potential, and carbonylated protein levels were evaluated. RESULTS: Hind limb unloading caused a loss of soleus muscle weight and muscle force. Intake of tea catechins significantly inhibited the unloading-induced decrease in force in isolated soleus muscle by 19% compared with the control group, although tea catechins did not affect muscle weight. In addition, intake of tea catechins suppressed the decrease in antioxidant potential and the increase in carbonyl myofibrillar protein. CONCLUSION: Ingestion of tea catechins minimized contractile dysfunction in skeletal muscle and muscle atrophy in unloaded muscle. This effect might be partly due to the lower oxidative modification of myofibrillar protein through the antioxidant activity of tea catechins. PMID- 21641775 TI - Microstructural alterations of the arcuate fasciculus in schizophrenia patients with frequent auditory verbal hallucinations. AB - Auditory verbal hallucinations (AVH) is a common and stressful symptom of schizophrenia. Disrupted connectivity between frontal and temporo-parietal language areas, giving rise to the misattribution of inner speech, is speculated to underlie this phenomenon. Disrupted connectivity should be reflected in the microstructure of the arcuate fasciculi (AF); the main connection between frontal and temporo-parietal language areas. In this study we compared microstructural properties of the AF and three other fiber tracts (cortical spinal tract, cingulum and uncinate fasciculus), between 44 schizophrenia patients with chronic severe hallucinations and 42 control subjects using diffusion tensor imaging (DTI) and magnetic transfer imaging (MTI). The DTI scans were used to compute fractional anisotropy (FA) and to reconstruct the fiber bundles of interest, while the MTI scans were used to compute magnetic transfer ratio (MTR) values. The patient group showed a general decrease in FA for all bundles. In the arcuate fasciculus this decreased FA was coupled to a significant increase in MTR values. A correlation was found between mean MTR values in both arcuate fasciculi and the severity of positive symptoms. The combination of decreased FA and increased MTR values observed in the arcuate fasciculi in patients suggests increased free water concentrations, probably caused by degraded integrity of the axons or the supportive glia cells. This suggests that disintegrated fiber integrity in the connection between frontal and temporo-parietal language areas in the schizophrenia patients is associated with their liability for auditory verbal hallucinations. PMID- 21641776 TI - The effect of adjunctive armodafinil on cognitive performance and psychopathology in antipsychotic-treated patients with schizophrenia/schizoaffective disorder: a randomized, double-blind, placebo-controlled trial. AB - BACKGROUND: The efficacy, safety and tolerability of adjunctive armodafinil for cognitive performance, and negative and affective symptoms, were examined in 60 patients with schizophrenia or schizoaffective disorder. METHOD: This was a 6 week, double-blind, placebo-controlled, fixed dose trial of armodafinil (150 mg/d) augmentation in patients with clinically stable schizophrenia or schizoaffective disorder. Cognition, psychopathology, alertness/wakefulness and adverse effects were assessed with standardized rating instruments. The primary endpoint was performance on measures of attention/vigilance. RESULTS: Patients were randomly allocated to adjunctive armodafinil or placebo. There was a significant Drug*Time interaction effect for attention/vigilance, due to modest non-significant worsening in the armodafinil group and improvement in the armodafinil group [CPT-Pairs d', F(1,40)=6.2, p=0.017]. However, it became non significant after correction for multiple comparisons. There were no differences between armodafinil and placebo in other cognitive domains or psychopathology measures. However, armodafinil was associated with significant improvement in the Scale for the Assessment of Negative Symptoms (SANS) anhedonia-asociality [F(1,41)=4.1, p=0.05]. CONCLUSIONS: There were no significant differences in neurocognitive measures between adjunctive armodafinil and placebo in this 6-week study. Armodafinil improved anhedonia-asociality, but not other negative symptom domains. PMID- 21641777 TI - Setting new standards in schizophrenia outcomes: symptomatic remission 3 years before versus after the Andreasen criteria. AB - Symptomatic remission is often mentioned as one of the treatment goals for schizophrenia. However, the consistently with which this is documented in the schizophrenia literature since the introduction of the consensus criteria proposed by Andreasen and colleagues in 2005 has yet to be investigated. Similarly, additional treatment goals which are being discussed include improved functioning and quality of life, but whether these goals are being increasingly documented in the literature alongside symptomatic remission is as yet unknown. The objective of this article is therefore to review the use of the term 'remission' in the schizophrenia literature from Europe, U.S.A. and the rest of the world from 2002 to 2007, before and after the introduction of the Andreasen criteria. A second objective is to determine whether these manuscripts documenting symptomatic remission are also addressing other concepts such as functioning, quality of life and relationships. This literature review indicates that the use of the Andreasen criteria is indeed increasing, although there are manuscripts documenting alternative remission criteria or using the term remission without documentation of specific remission criteria. From 2004 to 2007 the number of manuscripts mentioning remission without documenting specific criteria has fallen by approximately 50%. Within these manuscripts there is increasing awareness of functioning and quality of life as outcome measures, in particular in manuscripts generated in Europe and the U.S.A. This review highlights the growing importance of co-assessment of symptomatic remission and functional outcomes, and calls for further consideration of the most appropriate and consistent way to evaluate functioning of schizophrenia patients. PMID- 21641778 TI - Suitability of different media for in vitro cultivation of the ruminal protozoa species Entodinium caudatum, Eudiplodinium maggii, and Epidinium ecaudatum. AB - Three protozoal cultivation media were tested to determine the medium which best facilitated growth and viability of key B-type ciliates isolated from the sheep rumen. Entodinium caudatum and Eudiplodinium maggii were grown anaerobically in 50-ml flasks for 32 days in Caudatum-type (C), Kisidayova (K) or Dehority (M) medium. On day 32, in media K and M, E. caudatum cell counts were high with 5.6*10(3) and 7.8*10(3) mL(-1), respectively, and the proportion of dead cells was low with 0.6 and 1.4%, respectively. E. maggii concentrations when grown in medium M and C were 2.7*10(3) and 2.4*10(3) mL(-1), respectively, with 3.9 and 14.1% dead cells. Medium M, which favoured growth of both protozoa species, was tested again and Epidinium ecaudatum was included. Protozoa were grown for a 4 month period and samples were taken in the last two months on days 1, 7, 35 and 57. Average cell concentrations were 10.0, 0.8 and 0.5*10(3) mL(-1) for E. caudatum, E. maggii, and E. ecaudatum, respectively. In conclusion, medium M would appear to be the best choice for cultivating these three species in one medium. PMID- 21641779 TI - Intestinal ciliate composition found in the feces of the Turk rahvan horse Equus caballus, Linnaeus 1758. AB - Species composition and distribution of large intestinal ciliates were investigated in the feces from 15 Turk rahvan horses, living in the vicinity of Izmir, Turkey. Twenty-two ciliate genera consisting of 36 species were identified. This is the first report on intestinal ciliates in Turk rahvan horses and no previously unknown species were observed. The mean number of ciliates was 14.2+/-13.9*10(4) cells ml(-1) of feces and the mean number of ciliate species per host was 9.9+/-7.1. No ciliates were observed in 2 horses. Bundleia and Blepharocorys were considered to be the major genera since these ciliates were constantly found in high proportions. In contrast, Paraisotricha, Didesmis and Gassovskiella were only observed at low frequencies. The ciliates found in this survey had almost the same characteristics as those described in previous reports, suggesting that there was no significant geographic variation in the intestinal ciliate fauna of equids. PMID- 21641781 TI - Intelligent dental training simulator with objective skill assessment and feedback. AB - OBJECTIVE: We present a dental training simulator that provides a virtual reality (VR) environment with haptic feedback for dental students to practice dental surgical skills in the context of a crown preparation procedure. The simulator addresses challenges in traditional training such as the subjective nature of surgical skill assessment and the limited availability of expert supervision. METHODS AND MATERIALS: We identified important features for characterizing the quality of a procedure based on interviews with experienced dentists. The features are patterns combining tool position, tool orientation, and applied force. The simulator monitors these features during the procedure, objectively assesses the quality of the performed procedure using hidden Markov models (HMMs), and provides objective feedback on the user's performance in each stage of the procedure. We recruited five dental students and five experienced dentists to evaluate the accuracy of our skill assessment method and the quality of the system's generated feedback. RESULTS: The experimental results show that HMMs with selected features can correctly classify all test sequences into novice and expert categories. The evaluation also indicates a high acceptance rate from experts for the system's generated feedback. CONCLUSION: In this work, we introduce our VR dental training simulator and describe a mechanism for providing objective skill assessment and feedback. The HMM is demonstrated as an effective tool for classifying a particular operator as novice-level or expert-level. The simulator can generate tutoring feedback with quality comparable to the feedback provided by human tutors. PMID- 21641780 TI - Morphological and molecular phylogeny of dileptid and tracheliid ciliates: resolution at the base of the class Litostomatea (Ciliophora, Rhynchostomatia). AB - Dileptid and tracheliid ciliates have been traditionally classified within the subclass Haptoria of the class Litostomatea. However, their phylogenetic position among haptorians has been controversial and indicated that they may play a key role in understanding litostomatean evolution. In order to reconstruct the evolutionary history of dileptids and tracheliids, and to unravel their affinity to other haptorians, we have used a cladistic approach based on morphological evidence and a phylogenetic approach based on 18S rRNA gene sequences, including eight new ones. The molecular trees demonstrate that dileptids and tracheliids represent a separate subclass, Rhynchostomatia, that is sister to the subclasses Haptoria and Trichostomatia. The Rhynchostomatia are characterized by a ventrally located oral opening at the base of a proboscis that carries a complex oral ciliature. We have recognized two orders within Rhynchostomatia. The new order Tracheliida is monotypic, while the order Dileptida comprises two families: the new, typically bimacronucleate family Dimacrocaryonidae and the multimacronucleate family Dileptidae. The Haptoria evolved from the last common ancestor of the Litostomatea by polarization of the body, the oral opening locating more or less apically and the oral ciliature simplifying. The Trichostomatia originated from a microaerophylic haptorian by further simplification of the oral ciliature, possibly due to an endosymbiotic lifestyle. PMID- 21641782 TI - The general use of glycated haemoglobin for the diagnosis of diabetes and other categories of glucose intolerance: still a long way to go. AB - Glycated haemoglobin (HbA(1c)) is considered the 'gold standard' for monitoring metabolic control in diabetes. An International Expert Committee recently recommended HbA(1c) as a better method than measurement of glucose to use in the diagnosis of diabetes, based on its strong association with microvascular complications, a lower day-to-day variability and ease of use, not necessarily in the fasting state. These recommendations have been embraced by the American Diabetes Association (ADA), which stated in its Standards of Medical Care in Diabetes 2010 that "A(1c), fasting plasma glucose or the 2 h 75 g oral glucose tolerance test (OGTT) are appropriate for testing diabetes and assessing the risk of future diabetes," and that "a confirmed A(1c) >= 6.5% is diagnostic for diabetes." Measuring HbA(1c) has several advantages over glucose measurements, but its exclusive use should only be considered if the test is conducted under standardised conditions and its limitations are taken into due account. The impact of its use on the epidemiology of diabetes and other categories of glucose intolerance, as seen from recent reports, is also discussed. PMID- 21641783 TI - An in vivo and in vitro study on the protective effects of N-acetylcysteine on mitochondrial dysfunction in isoproterenol treated myocardial infarcted rats. AB - Altered mitochondrial function plays an important role in the pathology of myocardial infarction. We investigated the protective effects of N-acetylcysteine on mitochondrial dysfunction in isoproterenol induced myocardial infarcted rats. Rats were pretreated with N-acetylcysteine (10 mg/kg) orally daily for 14 days. After pretreatment, rats were induced myocardial infarction by isoproterenol (100 mg/kg) at an interval of 24 h for 2 days. Lipid peroxidation products, antioxidants, lipids, mitochondrial marker enzymes and calcium in the mitochondrial heart were determined. Transmission electron microscopic and in vitro studies were also done. Isoproterenol treatment caused significant increase in mitochondrial lipid peroxides and lipids except phospholipids with significant decrease in mitochondrial antioxidants. Significant decreased activities of marker enzymes and significant increased calcium were observed in mitochondria of myocardial infarcted rats. Pretreatment with N-acetylcysteine showed significant protective effects on all the biochemical parameters and preserved the integrity of heart tissue and restored normal mitochondrial function in myocardial infarcted rats. Transmission electron microscopic findings on the structure of the heart mitochondria confirmed the protective effects and in vitro study also confirmed the antioxidant potential of NAC. The possible mechanism for the improved cardiac mitochondrial function might be due to scavenging free radicals, improving the antioxidant and mitochondrial marker enzymes, maintaining GSH levels, lipids and Ca(2+) levels by its antioxidant effect. Thus, N acetylcysteine protected the mitochondrial heart from ISO treated mitochondrial damage. A diet containing N-acetylcysteine may be beneficial to myocardial infarcted heart. PMID- 21641784 TI - Museums and disease: using tissue archive and museum samples to study pathogens. AB - Molecular studies of archival and fossil samples have traditionally focused on the nucleic acids derived from the host species. However, there has recently been an increase in ancient DNA research on the identification and characterization of infectious agents within the hosts. The study of pathogens from the past provides great opportunities for discovering the causes of historical infection events, characterizing host-microorganism co-evolution and directly investigating the evolution of specific pathogens. Several research teams have been able to isolate and characterize a variety of different bacterial, parasite and viral microorganisms. However, this emerging field is not without obstacles. The diagenetic processes that make ancient DNA research generally difficult are also impediments to ancient pathogen research and perhaps more so given that their DNA may represent an even rarer proportion of the remaining nucleic acids in a fossil sample than host DNA. However, studies performed under controlled conditions and following stringent ancient DNA protocols can and have yielded reliable and often surprising results. This article reviews the advantages, problems, and failures of ancient microbiological research. PMID- 21641785 TI - Prompt correction of endotracheal tube positioning after intubation prevents further inappropriate positions. AB - STUDY OBJECTIVE: To determine whether the timely correction of endotracheal tube (ETT) positioning prevents further inappropriate positions. DESIGN: Prospective crossover study. SETTING: University-affiliated hospital. PATIENTS: 44 adult, ASA physical status 1, 2, and 3 patients undergoing open or laparoscopic abdominal procedures. INTERVENTIONS: ETT positioning was verified by both auscultation and fiberoptic bronchoscopy (FOB), after intubation, and before extubation. In laparoscopic procedures, two additional measurements were performed: after maximal abdominal gas insufflation and with head-down position. An ETT in the bronchus or at the carina was considered an inappropriate placement. An ETT <= one cm from the carina was considered a critical placement. MEASUREMENTS: The frequency of inappropriate and critical ETT positioning with both auscultation and FOB and the number of ETTs that remained in an incorrect position despite repositioning. MAIN RESULTS: FOB detected 5 inappropriately positioned ETTs, 4 of which were also detected by chest auscultation (P = 0.99). Critical positioning was detected by FOB in 6 patients, three of which were also detected by auscultation (P = 0.24). There were 15 other "out-of-desired range" positions (out of the 3-5 cm range) - one placed too high and 14 placed too low, while 18 were placed within the range of positions. All patients with inappropriate ETT positioning were women (P = 0.005). Age, body mass index, Mallampati grade > 3, thyromental distance < 6 cm, or laryngoscopy grade >= 2 were not associated with either inappropriate or critical placement. No episodes of inappropriate or critical positioning were detected by FOB or auscultation at the end of surgery. CONCLUSIONS: Early detection and prompt correction of inappropriate ETT positioning after intubation prevented further ETT migration into undesired positions. PMID- 21641786 TI - An advanced dual labeled gold nanoparticles probe to detect Cryptosporidium parvum using rapid immuno-dot blot assay. AB - The zoonotic protozoan parasite Cryptosporidium parvum poses a significant risk to public health. Due to the low infectious dose of C. parvum, remarkably sensitive detection methods are required for water and food industries analysis. However PCR affirmed sensing method of the causative nucleic acid has numerous advantages, still criterion demands for simple techniques and expertise understanding to extinguish its routine use. In contrast, protein based immuno detecting techniques are simpler to perform by a commoner, but lack of sensitivity due to inadequate signal amplification. In this paper, we focused on the development of a mere sensitive immuno detection method by coupling anti-cyst antibody and alkaline phosphatase on gold nanoparticle for C. parvum is described. Outcome of the sensitivity in an immuno-dot blot assay detection is enhanced by 500 fold (using conventional method) and visually be able to detect up to 10 oocysts/mL with minimal processing period. Techniques reported in this paper substantiate the convenience of immuno-dot blot assay for the routine screening of C. parvum in water/environmental examines and most importantly, demonstrates the potential of a prototype development of simple and inexpensive diagnostic technique. PMID- 21641787 TI - Film electrode prepared from oppositely charged silicate submicroparticles and carbon nanoparticles for selective dopamine sensing. AB - Film electrodes prepared from oppositely charged silicate submicroparticles and carbon nanoparticles was applied for selective dopamine sensing. Mesoporous silicate submicroparticles with tetraalkylammonium functionalities were prepared by sol-gel method. They were immobilised on an indium tin oxide film surface together with phenylsulphonated carbon nanoparticles by layer-by-layer method: alternative immersion into their suspensions. As it is shown by scanning electron microscopy the obtained film is composed of silicate submicroparticles covered by carbon nanoparticles. The nanoparticulate film is stable and its electroactive surface is significantly larger than substrate. Accumulation of redox active cations indicates that only fraction charged functionalities of carbon nanoparticles are employed in film formation. The obtained electrode exhibits catalytic properties towards dopamine oxidation and its interferences as ascorbic acid, uric acid and acetaminophen. This allows for selective determination of tenth micromolar concentration of dopamine in the presence of these interferences at milimolar level. The detection limit and linear range were determined to 0.1 * 10-6 mol dm-3 and 0.3-18 * 10-6 mol dm-3 respectively. PMID- 21641788 TI - Synthesis of a novel polyurethane-based-magnetic imprinted polymer for the selective optical detection of 1-naphthylamine in drinking water. AB - The first polyurethane based magnetic-MIP for the selective detection of 1 naphthylamine (1-NA) in drinking water has been synthesised. The synthesis has been carried out in a two-step process: first,the incorporation of magnetite coated-oleic acid nanoparticles (-Fe3O4-OA) into a lipophilic polymeric matrix (poly-MMA-co-EDMA) and second, the encapsulation of these magnetic seeds into the MIP structure by precipitation polymerisation. The mag-MIP was first RHTEM imaged showing a well-organised material with magnetite within the material and the imprinted polymer coating the magnetic core. Thereafter,it was evaluated by batch rebinding analysis and the derived Freundlich isotherm, calculating the number of binding sites (N(K(min)-K(max))=2.63 and 0.79 mmol g-1, for mag-MIP and mag-NIP, respectively)and apparent average adsorption constant (K(K(min)-K(max))=3.31 and 3.06 mmol-1, for mag-MIP and mag-NIP, respectively) showing a very effective imprinting process.We have also developed a magnetic optical sensor MIP by using an optical fiber coupled with a magnetic separator. An unexpected selectivity for 1-NA was revealed allowing the detection of this molecule in water, even in the presence of 4 structurally related compounds (2-naphthylamine, 1-naphthol, 2 naphthol and 1-naphthalenemethylamine), with a low limit of detection (LOD) = 18 ng mL-1. Finally, we applied this new hybrid material to the analysis of 1-NA in tap and mineral waters, obtaining a 91.6%average recovery rate. PMID- 21641789 TI - Metatarsal bars more effective than metatarsal pads in reducing impulse on the second metatarsal head. AB - BACKGROUND: The second metatarsal head is commonly involved in cases of metatarsalgia. As part of the conservative treatment, metatarsal bars and metatarsal pads are often prescribed. OBJECTIVE: To compare the effectiveness of metatarsal bars and metatarsal pads in reducing impulse on the second metatarsal head. METHOD: Thirty-five healthy subjects were monitored with an insole scanning system during walking in four different conditions: (a) wearing shoes only, (b) shoes plus metatarsal pads and shoes plus metatarsal bars, placed either (c) perpendicular to the foot axis or (d) oblique to the foot axis. The impulse under the second metatarsal head was measured using the first condition as a control. Both feet were examined in each subject resulting in a total of 840 measurements. RESULTS: Both metatarsal bars and metatarsal pads were effective in reducing impulse when compared with the control (P<0.01). Metatarsal bars were found to be more effective in reducing impulse as compared to the metatarsal pads (P<0.01), and the oblique position of the bars was more effective than the perpendicular one (P<0.01). CONCLUSIONS: The greatest reduction of impulse on the second metatarsal head in healthy subjects is achieved with the use of metatarsal bars in an oblique position. PMID- 21641791 TI - Use of thromboelastography to guide thromboprophylaxis after caesarean section. AB - BACKGROUND: Thromboprophylaxis is commonly required following caesarean section. However the effect of thromboprophylactic dosages of subcutaneous heparin on coagulation is unknown because conventional laboratory tests are largely unaffected. The aim of this study was to determine if thromboelastography could detect and quantify the effect of unfractionated heparin on coagulation profile when given at the time of surgery. METHODS: Nineteen women undergoing elective caesarean section were recruited. Blood samples collected before and after administration of subcutaneous unfractionated heparin 7500 IU underwent thromboelastography using both plain and heparinase cuvettes. Anti-factor Xa levels were also measured. RESULTS: There was a significant difference in R times between plain and heparinase samples (-10.6%, P=0.0072) indicating that thromboelastography could detect an effect of unfractionated heparin. Compared to baseline there were significant decreases of R times in plain (-20.4%, P=0.033) and heparinase (-28.8%, P=0.0001) samples despite the administration of unfractionated heparin. Anti-factor Xa levels were virtually undetectable (mean 0.01 U/mL). CONCLUSION: Thromboelastography was able to detect and quantify the effect of unfractionated heparin on blood coagulability, an effect not detected by conventional laboratory tests. Thromboelastography demonstrated a pro coagulant effect of surgery that was only partially mitigated by the use of unfractionated heparin. In this study, at a dose of 7500 IU subcutaneous unfractionated heparin appears to have little anticoagulant effect. PMID- 21641792 TI - A retrospective survey of adverse maternal and neonatal outcomes for parturients with congenital heart disease. AB - BACKGROUND: Parturients with congenital heart disease are at increased risk of maternal cardiac and neonatal complications. There is a paucity of literature regarding the relationship of complications with the type of anesthesia or mode of delivery. METHODS: We retrospectively reviewed all parturients with congenital heart disease undergoing delivery over a 7-year period at Tokyo Women's Medical University, Maternal and Perinatal Center to identify maternal cardiac and neonatal complications occurring during the peripartum period. RESULTS: Of 151 pregnancies in 128 women with congenital heart disease, there were 84 vaginal and 67 cesarean deliveries. Cesarean deliveries were performed with either neuraxial (n=51) or general (n=16) anesthesia. There were no maternal deaths and two neonatal deaths (one vaginal; one cesarean delivery). The incidence of maternal cardiac events was 1 in 84 (1%) for vaginal deliveries and 10 in 67 (15%) for cesarean deliveries. Neonatal complications occurred in 11 of 84 (13%) pregnancies with vaginal delivery and 25 of 67 (37%) pregnancies with cesarean delivery. Twenty-three elective cesarean deliveries occurred for maternal cardiac problems and were associated with a significant incidence of maternal cardiac (35%) and neonatal (65%) complications. The incidence of maternal cardiac events during delivery, when stratified by severity of cardiac disease, was similar to a previously derived cardiac risk index for pregnant women with cardiac disease. CONCLUSION: Despite a low overall incidence of maternal and neonatal mortality, pregnancy in women with congenital heart disease was associated with significant maternal cardiac and neonatal complications. Elective cesarean delivery with neuraxial anesthesia was a common approach for high-risk parturients with congenital heart disease; however, the benefit of this mode of delivery and anesthetic technique could not be ascertained. PMID- 21641793 TI - A selectionist account of de novo action learning. AB - How are novel actions generated and learned? We introduce a selectionist view of de novo action learning, and present some of the main postulates of such a view. This view contrasts with the notion that all actions are generated in response to particular stimuli and hence instructed by the world. It postulates that actions are generated in the actor (the organism) and selected by the environment (stimuli). Selection may occur iteratively until actions can be executed more rapidly and precisely, less variably, and eventually be elicited by particular stimuli. We also discuss experiments that support the particular predictions of this theory. PMID- 21641794 TI - Cell types, circuits, computation. AB - How does the connectivity of a neuronal circuit, together with the individual properties of the cell types that take part in it, result in a given computation? We examine this question in the context of retinal circuits. We suggest that the retina can be viewed as a parallel assemblage of many small computational devices, highly stereotypical and task-specific circuits afferent to a given ganglion cell type, and we discuss some rules that govern computation in these devices. Multi-device processing in retina poses conceptual problems when it is contrasted with cortical processing. We lay out open questions both on processing in retinal circuits and on implications for cortical processing of retinal inputs. PMID- 21641795 TI - Genetic polymorphisms in DNA repair and oxidative stress pathways associated with malignant melanoma susceptibility. AB - BACKGROUND: Base excision repair (BER) and nucleotide excision repair (NER) pathways eliminate a wide variety of DNA damage, including UV photoproducts. The ability of each individual to repair DNA damage following different causes might explain at least in part the variability in cancer susceptibility. Moreover, inflammatory response to UV exposure may further contribute to skin carcinogenesis by oxidative stress mechanisms. Single nucleotide polymorphisms in genes encoding various DNA-repair enzymes and oxidative stress factors may be candidate low-penetrance variants with a role in susceptibility to different cancers, particularly in those with aetiologies linked to environmental exposure, such as malignant melanoma (MM). METHODS: In this case-control study, 684 Spanish sporadic MM patients and 406 cancer-free control subjects were included and the role of 46 polymorphisms belonging to 16 BER and NER genes as well as 11 genes involved in oxidative stress processes were investigated. RESULTS: One polymorphism was identified to be individually associated with MM in the Spanish population. The variant was found in the NOS1 oxidative stress gene (rs2682826; p value=0.01). These results suggest a putative role of oxidative stress processes in the genetic predisposition to melanoma. CONCLUSION: To the authors' knowledge, this is the largest DNA repair-related SNP study in melanoma risk conducted in the Spanish population up to now. Furthermore, it also represents a comprehensive genetic study of several oxidative stress polymorphisms tested in relation to MM susceptibility. PMID- 21641796 TI - Design, synthesis and biological activity of novel peptidyl benzyl ketone FVIIa inhibitors. AB - Herein is described the synthesis of a novel class of peptidyl FVIIa inhibitors having a C-terminal benzyl ketone group. This class is designed to be potentially suitable as stabilization agents of liquid formulations of rFVIIa, which is a serine protease used for the treatment of hemophilia A and B inhibitor patients. A library of compounds was synthesized with different tripeptide sequences, N terminals and d-amino acids in the P3 position. Cbz-D-Phe-Phe-Arg-bk (33) was found to be the best candidate with a potency of K(i)=8MUM and no substantial inhibition of related blood coagulation factors (thrombin and FXa). Computational studies revealed that 33 has a very stable binding conformation due to intramolecular hydrogen bonds, which cannot be formed with l-Phe in the P3 position. Nonpolar amino acids were found to be superior, probably due to a minimization of the cost of desolvation upon binding to FVIIa. PMID- 21641797 TI - Synthesis and screening of (E)-1-(beta-D-galactopyranosyl)-4-(aryl)but-3-ene-2 one against Mycobacterium tuberculosis. AB - A series of galactose-derived aryl enones were synthesised and screened against Mycobacterium tuberculosis H(37)Rv. Preliminary results were promising with MIC values in the range 1.56-12.5 MUg/mL. PMID- 21641798 TI - Synthesis of new opioid derivatives with a propellane skeleton and their pharmacology: part 1. AB - The observation that 17-cyclopropylmethylmorphinan derivatives without the 4,5 epoxy ring showed more kappa selectivity than those with a 4,5-epoxy ring led us to develop a working hypothesis: the position of the plane composed of the A and B rings would influence the opioid receptor type selectivity and that the decrease in the torsion angle C11-C12-C13-C14 could improve the kappa selectivity. Consistent with our hypothesis, KNT-42 with an N-cyclopropylmethyl propellane structure, whose A and B rings were fixed in a torsion angle of approximately 0 degrees , showed kappa selective agonist activity. PMID- 21641799 TI - Synthesis and antimycobacterial activity of highly functionalized tetrahydro 4(1H)-pyridinones. AB - A series of 35 2e,3e,6e-triaryltetrahydro-4(1H)-pyridinones, 2e,3e,5e,6e tetraaryltetrahydro-4(1H)-pyridinones and their N-nitroso and N-cyano analogs have been prepared. All these 35 compounds obtained were screened for their in vitro activity against Mycobacterium tuberculosis H37Rv (MTB). Among them, the N nitrosopyridinones are found to be more active against MTB than the corresponding N-CN analogs, which, in turn, were slightly more active than NH analogs. In particular, the N-nitroso compounds, 3d, 4b and 4e with halogen-bearing phenyl rings at 2,6-positions showed maximum activity with MIC values of 3.97, 3.11 and 3.11 MUM, being more efficacious than the first line anti-TB drugs, ciprofloxacin, ethambutol and pyrazinamide. A general trend has also been discerned in all the three classes of NH, N-CN and N-NO compounds, in each of which those bearing four aryl rings display higher activity than that having three analogously substituted aryl rings disclosing that lipophilicity could be an important factor underlying antimycobacterial activity. PMID- 21641800 TI - 173rd ENMC International Workshop: congenital muscular dystrophy outcome measures 5-7 March 2010, Naarden, The Netherlands. PMID- 21641801 TI - Spectrum of breast disease encountered in HIV-positive patients at a community teaching hospital. AB - INTRODUCTION: HIV infection directly and indirectly affects breast tissue. This study describes the spectrum of HIV-related breast disease encountered at a community teaching hospital. METHODS: A 9 year retrospective review was performed of HIV-positive patients with a breast-related diagnosis seen at our institution. Patient demographics, HIV status, comorbid disease, medications, clinical findings, diagnostic procedure, pathology, treatment and outcome were recorded. RESULTS: A total of 46 individuals were included with a median age of 47 years (range 24-64 years) and male:female ratio of 1:3 (12 men and 34 women). Mean duration of HIV infection was 7 years during which time 46% of patients had an AIDS defining illness. Median CD4 cell count was 437 cells/mm(3) (range 2 to >=500 cells/mm(3)) at the time of the breast diagnosis. Breast disease identified included benign conditions (59% total: 92% for men, 47% for women), infection (17% total: 8% for men, 21% for women), cancer (22% total: 0% for men, 29% for women), and atypia (2% total: 0% for men, 3% for women). Patients with a breast infection had a lower median CD4 cell count than those with breast cancer or benign conditions. Gynecomastia was detected in seven out of 12 (58%) men. In these men, antiretroviral therapy (ART) of all drug classes was associated with gynecomastia. Breast cancer occurred only in women and included patients with invasive ductal carcinoma (n = 7), ductal carcinoma in situ (n = 2), and liposarcoma diagnosed in one individual. Specific risk factors for breast cancer in this setting were not identified. Five (11%) patients died, only one from breast disease during the study period. CONCLUSION: These data indicate that increased longevity in patients with chronic HIV infection may be associated with the occurrence of breast conditions in both men and women. A broad spectrum of breast disease should be anticipated in HIV-infected persons living longer with effective ART. PMID- 21641802 TI - Radiofrequency ablation of early breast cancer followed by delayed surgical resection--a promising alternative to breast-conserving surgery. AB - To examine the radiofrequency ablation (RFA) reliability in early breast cancer, we performed RFA followed by delayed surgical resection on 41 patients with invasive or non-invasive breast carcinoma less than 2 cm. MRI scans were obtained before ablation and resection. Excised specimens were examined pathologically by haematoxylin-eosin and nicotinamide adenine dinucleotide-diaphorase staining. 40 patients completed 1 RFA session, which was sufficient to achieve complete tumour cell death. Overall complete ablation rate was 87.8% (36/41). There were no treatment-related complications other than that of a superficial burn in 1 case. After RFA, the tumour was no longer enhanced on MRI in 25/26 (96.2%) cases. Residual cancer, which was suspected on MRI in 1 case, was confirmed pathologically. MRI could be an applicable modality to evaluate therapeutic effect. RFA could be an alternate local treatment option to breast-conserving surgery for early breast cancer. PMID- 21641803 TI - A sensitivity comparison of clinical tests for postural instability in patients with Huntington's disease. AB - Falls are a common complication in Huntington's disease (HD) and detection of postural instability (PI) may be useful for identifying patients who are at risk of falls. The aim of our study was to find the most sensitive clinical test for PI in patients with HD and to correlate PI with the other symptoms. 20 HD patients were examined using: (1) The Unified Huntington's Disease Rating Scale (UHDRS), (2) The Mini Mental State Examination (MMSE) and (3) six clinical tests for PI. Inter-scale and test correlations were inspected and uni-dimensionality, validity, and measurement precision were analyzed using a factor analysis model/latent PI score. PI was compared with information obtained from surveys completed independently by patients and caregivers, and both reports were evaluated for absolute agreement using intraclass correlations (ICCs). PI was found in 16 patients; the tests correlated better with caregivers' reports (r=0.78) than patients' responses. The validity of the patients' answers decreased with the level of MMSE scores. PI correlated with MMSE (r=0.64, p<0.01), the subscale score for voluntary movements (r=0.86, p<0.01), the overall motor subscore (r=0.73, p<0.01), and the Luria test subscore (r=0.87, p<0.01). The stance with feet close together and tandem gait correlated best with the latent PI score, and factor loadings confirmed they were the most sensitive tests for PI detection in HD. PMID- 21641804 TI - Emerging roles for Lys11-linked polyubiquitin in cellular regulation. AB - Polyubiquitin chains are assembled via one of seven lysine (Lys) residues or the N terminus. The cellular roles of Lys48- and Lys63-linked polyubiquitin have been extensively studied; however, the cellular functions of Lys11-linked chains are less well understood. Recent insights into Lys11-linked ubiquitin chains have revealed their important function in cell cycle control. Additionally, Lys11 linkages have been identified in the context of mixed chains in many other cellular pathways. In this review, we introduce the specific enzymes that mediate Lys11-linked chain assembly and disassembly, and discuss the diverse cellular processes in which Lys11 linkages participate. Notably, mechanistic insights have revealed how the E2 ubiquitin-conjugating enzyme UBE2S achieves its Lys11 linkage specificity, and two structures of Lys11-linked polyubiquitin highlight the dynamic nature of this compact chain type. PMID- 21641806 TI - Improved therapeutic effect of folate-decorated PLGA-PEG nanoparticles for endometrial carcinoma. AB - Folate (FOL) mediated poly-lactide-co-glycolide-polyethylene glycol nanoparticles (FOL-PEG-PLGA NPs) bearing paclitaxel (PTX) were prepared for the effective delivery of drug to endometrial carcinoma. The average size, zeta potential and encapsulation efficiency of FOL-targeted NPs were found to be around 220 nm, 30.43 mV and 95.6%. Cellular uptake was observed. The accumulation of FOL targeted NPs depends on dual effects of passive and active targeting. The FOL targeted PTX NPs showed a greater cytotoxicity against HEC-1A cancer cells in vitro and in vivo, which might be induced by apoptosis. H&E staining did not showed apparent tissue damage to liver and kidney of the mice after injecting NPs intravenously. These results suggest that the novel FOL-PEG-PLGA NPs could be a potential delivery system with excellent therapeutic efficacy for targeting the drugs to cancer cells. PMID- 21641805 TI - How dormant origins promote complete genome replication. AB - Many replication origins that are licensed by loading MCM2-7 complexes in G1 are not normally used. Activation of these dormant origins during S phase provides a first line of defence for the genome if replication is inhibited. When replication forks fail, dormant origins are activated within regions of the genome currently engaged in replication. At the same time, DNA damage-response kinases activated by the stalled forks preferentially suppress the assembly of new replication factories, thereby ensuring that chromosomal regions experiencing replicative stress complete synthesis before new regions of the genome are replicated. Mice expressing reduced levels of MCM2-7 have fewer dormant origins, are cancer-prone and are genetically unstable, demonstrating the importance of dormant origins for preserving genome integrity. We review the function of dormant origins, the molecular mechanism of their regulation and their physiological implications. PMID- 21641807 TI - Synthesis and biological evaluation of alpha-ketoamides as inhibitors of the Dengue virus protease with antiviral activity in cell-culture. AB - The development of small molecule inhibitors of the viral protease is of considerable interest for the treatment of emergent flaviviral diseases such as Dengue or West Nile fever. Until today little progress has been made in finding drug-like compounds that inhibit the protease and provide a starting point for lead optimization. We describe here the initial steps of a drug discovery effort that focused on the styryl pharmacophore, combined with a ketoamide function to serve as electrophilic trap for the catalytic serine. This resulted in a fragment like lead compound with reasonable target affinity and good ligand efficiency, which was extensively modified to explore structure-activity relationships. Selected compounds were cross-tested against the West Nile virus protease and thrombin, indicating that selectivity for one or more flaviviral proteases can be achieved. Finally, the antiviral activity of several protease inhibitors was confirmed in a cell-culture model of Dengue virus replication. The SAR presented here may serve as starting point for further drug discovery efforts with the aim of targeting flaviviral proteases. PMID- 21641808 TI - New vasorelaxant indole alkaloids, villocarines A-D from Uncaria villosa. AB - Villocarines A-D (1-4), four new indole alkaloids have been isolated from the leaves of Uncaria villosa (Rubiaceae) and their structures were elucidated by 2D NMR methods and chemical correlations. Villocarine A (1) showed vasorelaxation activity against rat aortic ring and showed inhibition effect on vasocontraction of depolarized aorta with high concentration potassium, and also inhibition effect on phenylephrine (PE)-induced contraction in the presence of nicardipine in a Ca(2+) concentration-dependent manner. The vasorelaxant effect by 1 might be attributed mainly to inhibition of calcium influx from extracellular space through voltage-dependent calcium channels (VDC) and/or receptor-operated Ca(2+) channels (ROC), and also partly mediated through the increased release of NO from endothelial cells and opening of voltage-gated K(+)-channels. PMID- 21641809 TI - Measurement of long lived radioactive impurities retained in the disposable cassettes on the Tracerlab MX system during the production of [18F]FDG. AB - Using a High- Purity Germanium gamma-ray spectrometer, a number of radioisotopes have been identified within Tracerlab MX radiochemistry system cassettes used to synthesise [18F]FDG. Twenty radiochemistry cassettes were measured and the average total activity of each radioisotope was determined. Using these values and decay correction, the minimum time the cassettes should be left in a decay store before the specific activity falls below 0.4B q/g, the limit for disposal alongside Clinical Waste was found to be 24 months. PMID- 21641810 TI - Design, development and characterization of multi-functionalized gold nanoparticles for biodetection and targeted boron delivery in BNCT applications. AB - The aim of this study is to optimize targeted boron delivery to cancer cells and its tracking down to the cellular level. To this end, we describe the design and synthesis of novel nanovectors that double as targeted boron delivery agents and fluorescent imaging probes. Gold nanoparticles were coated with multilayers of polyelectrolytes functionalized with the fluorescent dye (FITC), boronophenylalanine and folic acid. In vitro confocal fluorescence microscopy demonstrated significant uptake of the nanoparticles in cancer cells that are known to overexpress folate receptors. PMID- 21641811 TI - Salivary gland tumours: 25 years of experience from a single institution in Croatia. AB - INTRODUCTION: The aim of this study was to determine the types, frequency, distribution, and demographic characteristics of salivary gland tumours in a large representative sample. PATIENTS AND METHODS: We retrospectively analysed the medical records of 779 patients with tumours of the salivary glands surgically treated from 1985 to 2009 at a single institution. RESULTS: There were 500 benign and 279 malignant tumours. The average age of patients with benign tumours was 50 years and of malignant salivary gland tumours 56 years. No differences in age and incidence of tumours existed between males and females. The majority of the tumours occurred in the parotid gland (509), followed by the minor salivary glands (212), the submandibular gland (51) and lastly, the sublingual gland (7). Minor salivary gland tumours occurred most frequently on the palate, the pleomorphic adenoma being the most frequent benign tumour type and the adenoid cystic carcinoma being the commonest malignant tumour. Tumours of the sublingual gland were rare, but all were malignant. Malignant tumours were more common in the minor salivary glands and the submandibular gland. CONCLUSION: This large study of salivary gland tumours in Croatia could improve our understanding of the significant differences in the global distribution of salivary gland tumours which have been reported. PMID- 21641812 TI - Lateral rhinotomy combined with anterior transantral approach for the treatment of large malignant melanoma of the nasal cavity involving the nasopharynx. AB - The authors report a case of nasal malignant melanoma involving the septum, nasal turbinates and nasopharynx which was approached by lateral rhinotomy combined with an anterior transantral approach with excision of the septum and the lateral wall of the nasal cavity for wide exposure of the tumour. Using this technique complete surgical resection of a large nasal malignant melanoma was achieved with minimal morbidity and good aesthetics. PMID- 21641813 TI - Prediction of significant coronary artery disease in patients undergoing operations for rheumatic mitral valve disease. AB - OBJECTIVE: This study aims to develop a logistic regression model and a simple score system for the prediction of significant coronary artery disease (CAD) in patients undergoing operations for rheumatic mitral valve disease. METHODS: A total of 1241 rheumatic patients (mean age 57+/-6 years), who underwent routine coronary angiography (CAG) before mitral valve operations between 1998 and 2009, was analyzed. To identify low-risk (<=5%) patients, a bootstrap refined logistic regression model on the basis of clinical risk factors was developed, from which an additive model was derived. Receiver operating characteristic (ROC) curves were used to compare discrimination, and precision was quantified by the Hosmer Lemeshow statistic. Significant coronary atherosclerosis was defined as 50% or more luminal narrowing in one or more major epicardial vessels by means of CAG. RESULTS: One hundred twenty-seven (10.2%) patients had significant coronary atherosclerosis. Independent predictors of significant CAD include age, male sex, hypertension, angina, smoking, and hypercholesterolemia. Five hundred and fifty patients were designated as low risk according to our logistic regression and additive models. Of these patients, only 6 (1.1%) had single-vessel disease, and none had multivessel disease. Our models proved more efficient than established regression models. CONCLUSIONS: Our logistic regression model could estimate the risk of significant CAD in rheumatic patients undergoing mitral valve operations, while the additive simple score system could reliably identify the low-risk patients in whom routine preoperative angiography might be safely avoided. PMID- 21641814 TI - Temporary percutaneous right ventricular support using a centrifugal pump in patients with postoperative acute refractory right ventricular failure after left ventricular assist device implantation. AB - OBJECTIVE: Acute right ventricular (RV) failure is a life-threatening condition with a poor prognosis, and sometimes the use of mechanical circulatory support is inevitable. In this article, we describe our experience using a centrifugal pump as a temporary percutaneous right ventricular assist device (RVAD) in patients with postoperative acute refractory RV failure after left ventricular assist device (LVAD) implantation. METHODS: We retrospectively reviewed eight consecutive patients with acute RV failure who underwent temporary percutaneous RVAD implantation using a centrifugal pump after LVAD implantation between April 2008 and February 2011. A Dacron graft was attached to the main pulmonary artery and passed through a subxiphoid exit, where the outflow cannula was inserted. The inflow cannula was percutaneously cannulated using Seldinger's technique in the femoral vein. The chest was definitely closed. The technique allowed bedside removal, avoiding chest re-opening. RESULTS: The median patient age was 52 years (range: 41-58). The median duration of support was 14 days (range: 12-14). RV systolic function improved; central venous pressure and mean pulmonary artery pressure decreased significantly after RVAD support. In three patients, an oxygenator was integrated into the RVAD due to impaired pulmonary function. Six patients were successfully weaned. Five patients survived to hospital discharge. Technical problems or serious complications concerning decannulation were not observed. CONCLUSION: This report suggests that implantation of temporary percutaneous RVAD using a centrifugal pump is a safe alternative in the treatment of postoperative acute refractory RV failure. Ease of device implantation, weaning, explantation, and limited number of complications justify a liberal use. PMID- 21641815 TI - Optimization of photodynamic therapy response by survivin gene knockdown in human metastatic breast cancer T47D cells. AB - Photodynamic therapy (PDT) leads to the generation of cytotoxic oxygen species that appears to stimulate several different signaling pathways, some of which lead to cell death, whereas others mediate cell survival. In this context, we observed that PDT mediated by methyl-5-aminolevulinic acid as the photosensitizer resulted in over-expression of survivin, a member of the inhibitor of apoptosis (IAP) family that correlates inversely with patient prognosis. The role of survivin in resistance to anti-cancer therapies has become an area of intensive investigation. In this study, we demonstrate a specific role for survivin in modulating PDT-mediated apoptotic response. In our experimental system, we use a DNA vector-based siRNA, which targets exon-1 of the human survivin mRNA (pSil_1) to silence survivin expression. Metastatic T47D cells treated with both pSil_1 and PDT exhibited increased apoptotic indexes and cytotoxicity when compared to single-agent treated cells. The treatment resulted in increased PARP and caspase 3 cleavage, a decrease in the Bcl-2/Bak ratio and no participation of heat shock proteins. In contrast, the overexpression of survivin by a survivin-expressed vector increased cell viability and reduced cell death in breast cancer cells treated with PDT. Therefore, our data suggest that combining PDT with a survivin inhibitor may attribute to a more favorable clinical outcome than the use of single-modality PDT. PMID- 21641816 TI - Heavily calcified occlusive lesions of the iliac artery: long-term patency and CT findings after stent placement. AB - PURPOSE: To evaluate the influence of heavy calcification on iliac arterial stent expansion and patency and to define the spatial relationship between the stent and heavy calcifications on computed tomography (CT). MATERIALS AND METHODS: Thirteen patients (11 men, two women; mean age, 66.5 y) with 14 heavily calcified iliac arteries received primary stent treatment between 1998 and 2008. Anatomic success was defined by less than 30% residual stenosis on final follow-up CT angiography. Hemodynamic success was defined as an increase in the ankle-brachial index (ABI) of at least 0.15 versus baseline. Clinical success was defined by achievement of clinical improvement of at least one clinical category. Stent patency; anatomic, hemodynamic, and clinical success rate; morphology of heavy calcifications; calcium score; and stent geometry were evaluated. RESULTS: Stents were successfully inserted in all cases. During a mean follow-up of 33.6 months (range, 8-55 mo), the stent-implanted iliac arteries remained anatomically patent in all patients on final follow-up. The anatomic, hemodynamic, and clinical success rates were 28.6%, 60%, and 78.6%, respectively. Mean ABIs were 0.68 +/- 0.22 before the procedure and 0.91 +/- 0.23 after the procedure (P = .021). Mean luminal stenosis measurements were 77.9% before the procedure and 47.9% after the procedure (P = .008). CONCLUSIONS: Iliac stents in heavily calcified lesions showed hemodynamically significant residual stenosis in a considerable number of cases. However, stent patency was not affected even with incomplete expansion of the stent. PMID- 21641817 TI - Does inpatient quality of care differ by age among US veterans with ischemic stroke? AB - BACKGROUND: Some studies have found that older individuals are not as likely as their younger counterparts to be treated with some guideline-based stroke therapies. We examined whether age-related differences in inpatient quality of care exist among US veterans with ischemic stroke. METHODS: This was a retrospective study of a national sample of veterans admitted to 129 Veterans Affairs medical centers for ischemic stroke during fiscal year 2007. Inpatient stroke care quality was examined across 14 inpatient processes of care, including dysphagia screening, National Institutes of Health Stroke Scale (NIHSS) score documentation, thrombolysis, deep venous thrombosis prophylaxis, antithrombotic therapy by hospital day 2 and at discharge, early ambulation, fall risk assessment, pressure ulcer risk assessment, rehabilitation needs assessment, atrial fibrillation management, lipid management, smoking cessation counseling, and stroke education. RESULTS: Among the 3939 veterans with ischemic stroke, the mean age was 67.8 years (standard deviation, 11.5). The overall performance rate was >70% for 10 of the 14 quality indicators. In unadjusted analyses, older patients were less likely to receive lipid management, smoking cessation, NIHSS documentation, and early ambulation compared with younger patients; conversely, older patients were more likely to receive dysphagia screening and stroke education. After adjusting for demographic, clinical, and hospital level characteristics, the age-related differences in processes of care were less consistent; however, the youngest patients were more likely to receive smoking cessation counseling and the oldest patients were less likely to receive lipid management. CONCLUSIONS: Risk-adjusted inpatient stroke care quality varies little with age for veterans admitted to a Veterans Affairs medical center for acute ischemic stroke. PMID- 21641818 TI - Decrease in bispectral index in a patient undergoing coronary artery bypass graft surgery. PMID- 21641819 TI - Target-controlled dosing of remifentanil during cardiac surgery reduces postoperative hyperalgesia. AB - OBJECTIVE: One of the strategies to attenuate opioid-induced hyperalgesia (OIH) may be to decrease intraoperative doses of opioids by using target-controlled infusion (TCI). DESIGN: Double-blind and randomized study. SETTING: A single university hospital. PARTICIPANTS: Forty American Society of Anesthesiologists II to III patients scheduled for elective cardiac surgery. INTERVENTIONS: patients were randomized to 1 of the 2 groups: 1 group received an infusion of intraoperative remifentanil using TCI (target: 7 ng/mL), and the 2nd one was given an intraoperative continuous infusion (CI) (0.3 MUg/kg/min). The anesthestic protocol and postoperative pain management were the same in both groups. The extent of mechanical dynamic hyperalgesia on the middle line perpendicular to the wound was considered the primary endpoint. The secondary endpoints were other results of dynamic and punctuate hyperalgesia until postoperative day 7, visual analog scale (VAS) and verbal rating scale (VRS) scores, and total morphine consumption until postoperative day 2. MEASUREMENTS AND MAIN RESULTS: Morphometric and demographic characteristics and duration of surgery were comparable in both groups. Intraoperative remifentanil consumption was greater in CI than in TCI group (5,329 [1,833] v 3,662 [1,160] MUg, p = 0.003). During the first 44 hours, there were no differences in morphine consumption, VAS, and VRS. The extent of hyperalgesia was significantly lower on postoperative days 1, 2, and 4 in the TCI group than in the CI group on the 3 evaluated lines (p < 0.05). Punctuate hyperalgesia evaluating 3 different points was lower in the TCI than in the CI group from postoperative day 1 until postoperative day 7 (p < 0.05). CONCLUSIONS: The intraoperative decrease of opioid consumption when comparing the CI versus TCI mode of administration of remifentanil led to less OIH after cardiac surgery. PMID- 21641820 TI - Unexpected detection of internal jugular vein thrombus during ultrasound-guided central venous cannulation. PMID- 21641821 TI - Adjuvant therapy with methylene blue in the treatment of postoperative vasoplegic syndrome caused by carcinoid crisis after tricuspid valve replacement. PMID- 21641822 TI - The thoracicus longus and thoracodorsalis nerve block: a supplemental block for thoracic anesthesia analgesia. PMID- 21641823 TI - Being present in more than one place at a time? Patterns of mental self localization. AB - Research in cognitive neuroscience and spatial presence suggests that human mental self-localization is tied to one place at a given point in time. In this study, we examined whether it is possible to feel localized at two distinct places at the same time. Participants (N=30) were exposed to a virtual rollercoaster and they continuously judged to what extent they felt present in the immediate environment and in the mediated environment, respectively. The results show that participants distributed their self-localization to both environments, and the two values added up to closely 100% over time. In addition, even though the judgments are highly idiosyncratic, they were almost perfectly inversely related. This indicates that individuals can distribute their self over two distinct places. These findings provide important insights about understanding of the human self-localization. PMID- 21641824 TI - An unusual association of endomyocardial fibrosis and hypertrophic cardiomyopathy in a patient with heart failure. AB - A 69-year-old female patient presented heart failure with preserved ejection fraction and atrial fibrillation. Echocardiography and late gadolinium enhancement magnetic resonance imaging were suggestive of endomyocardial fibrosis (EMF). The patient underwent cardiac surgery, and after surgery, she developed low cardiac output syndrome and died. Postmortem examination revealed residual fibrosis of the left ventricle (LV), mild endocardial fibrous deposition of the right ventricle, and severe concentric, symmetrical LV hypertrophy. Histological examination of the surgically resected material from the LV confirmed EMF. Histopathology of the interventricular septum disclosed myocardial hypertrophy and disarray plus fine interstitial fibrosis, typical of hypertrophic cardiomyopathy. The present case illustrates the association of two different patterns of cardiomyopathies in the same patient. PMID- 21641825 TI - The use of the reverse shoulder arthroplasty for treatment of failed total shoulder arthroplasty. AB - BACKGROUND: This study evaluated the outcomes of patients with failed total shoulder arthroplasty (TSA) who were treated with conversion to reverse shoulder arthroplasty (RSA). MATERIALS AND METHODS: We performed a retrospective case series of 24 consecutive patients with failed TSA who were treated with conversion to RSA. Twenty-two patients (16 women, 6 men) had a minimum 2-year clinical and radiographic follow-up. The average age at the time of revision was 68 years (range, 51-84 years). Indications for conversion to RSA included failure of TSA from glenohumeral instability in 19, mechanical failure of the humeral or glenoid component in 10, and infection in 2. RESULTS: The median total American Shoulder and Elbow Surgeons score improved from 38.5 preoperatively to 67.5 (P < .001). Visual analog scale pain scores decreased from 5 to 1.5 (P < .001), and function improved from 2 to 6.5 (P < .001). The median Simple Shoulder Test improved from 1 to 5 (P = .006). Forward flexion improved from 50 degrees to 130 degrees (P < .001), abduction from 45 degrees to 100 degrees (P < .001), and external rotation from 12.5 degrees to 49.5 degrees (P = .056). Internal rotation improved from a spinal level of S2 to L3 (P = .064). Fourteen patients rated their outcome as excellent, 3 as good, 3 as satisfactory, and 2 as unsatisfactory. The overall complication rate was 22.7% (5 of 22). CONCLUSION: RSA can be an effective treatment for failed TSA by decreasing pain and improving shoulder function. However, RSA in the revision setting is associated with a higher complication rate. PMID- 21641826 TI - Retrospective comparison of titanium elastic nail (TEN) and reconstruction plate repair of displaced midshaft clavicular fractures. AB - BACKGROUND: Titanium elastic nails (TENs) are commonly used to repair displaced midshaft clavicular fractures. Although several clinical studies have shown that the use of TENs is more effective and less invasive than traditional plate fixation, high rates of complications associated with TENs have been reported. This retrospective study compared the use of TENs with reconstruction plates in the treatment of displaced midshaft clavicular fractures. MATERIALS AND METHODS: From January 2005 to July 2007, 141 patients with displaced midshaft clavicular fractures were treated with TEN or plate fixation. At 6 and 24 months postoperatively, we evaluated the patients' general health, operative complications, and functional recovery. RESULTS: The mean bone union time of TEN patients was 12.4 +/- 3.4 weeks, whereas that in the plate group was 14.4 +/- 3.7 weeks. The time of union was significantly shorter in the TEN group than that in the plate group. There was no significant difference between these 2 groups' rates of complications such as nonunion or malunion. Shoulder function scores were significantly better in the TEN group than in the plate group at 6 months postoperatively. There was no significant difference at 24 months postoperatively. Patients were more satisfied with the cosmetic appearance and overall outcome after TEN repair than after plate fixation. CONCLUSIONS: TEN fixation of displaced midshaft clavicular fractures allows for a faster functional recovery, higher patient satisfaction, and a more cosmetically satisfactory appearance than plate fixation. The complication rates of both TENs and reconstruction plates were similar. PMID- 21641827 TI - Interface micromotions increase with less-conforming cementless glenoid components. AB - BACKGROUND: The optimal degree of conformity between the glenoid and humeral components in total shoulder arthroplasty for best performance and durability is still a matter of debate. The main aim of this study is to evaluate the influence of joint conformity on the bone-implant interface micromotions in a cementless glenoid implant. MATERIALS AND METHODS: Polyethylene inlays with different degrees of conformity (radial mismatch of 0, 2, 4, and 6 mm) were mounted on a cementless metal back and then implanted in a bone substitute. These glenoid components were loaded by a prosthetic humeral head during a force-controlled experiment. Normal-to-interface micromotions and bone substitute deformations were measured at different points of the interface. Rim displacement and humeral head translation were also measured. A finite element (FE) model of the experiments was implemented to estimate the normal- and tangent-to-interface micromotions in the entire bone-implant interface. RESULTS: All measured variables increased with less-conforming PE inlays. Normal-to-interface micromotions were significantly larger (P < .05) when the radial mismatch was 6 mm compared with the fully conforming inlay. The FE model was in agreement and complemented the experimental results. FE model-predicted interface micromotions were already significantly larger when the radial mismatch was equal to 4 mm. DISCUSSION: In a force-controlled experiment with a cementless glenoid component, a non-conforming PE inlay allows larger interface micromotions than a conforming inlay, reaching a magnitude that may hamper local bone ingrowth in this type of component. This is mainly because of the larger humeral head translation that boosts the effects of the so-called rocking-horse phenomenon. PMID- 21641828 TI - Locked bucket-handle tear of the anterior labrum causing persistent subluxation of a dislocated shoulder. PMID- 21641829 TI - Incidence and implications of early postoperative wound complications after total elbow arthroplasty. AB - HYPOTHESIS: Other than an awareness, there is little detailed information regarding wound problems after total elbow arthroplasty. The purpose of this study was to (1) determine the incidence of wound complications after elbow arthroplasty, (2) document the long-term implications, (3) characterize risk factors, and (4) discuss a management strategy. We hypothesize that the incidence of this complication can be reduced with careful preoperative planning. MATERIALS AND METHODS: We reviewed 1749 total elbow arthroplasties. The average patient age was 61.5 years (range; 30-91 years). Wound complications were diagnosed according to the criteria of the Centers for Disease Control and Prevention. RESULTS: We identified and studied 97 patients (5.5%) from the 1749 procedures. The most common problems were delayed healing and drainage in 34 and wound hematoma in 33, of which 9 (27%) progressed to secondary deep infection. Of the 97 patients, 86 (88.7%) healed with the retention of the implant, 24 (~25%) progressed to a septic elbow, and 11 (~50%) required resection. Patients with rheumatoid arthritis represented 33% of the entire sample, but represented 45.8% of those with septic complications. Posttraumatic arthritis patients represented 58% of the entire sample and only 33% of those with septic problems (P < .05). CONCLUSIONS: The overall incidence of serious wound complications was slightly less than anticipated; however, the significance was considerable. Patients with rheumatoid arthritis are most vulnerable. Persistent wound drainage showed a high correlation for deep infection and subsequent implant removal. Anticipation of potential problems and appropriate prophylactic management may avoid wound complications. PMID- 21641830 TI - Arthroscopic tibiotalocalcaneal arthrodesis with intramedullary nail with fins: a case series. AB - Arthroscopic arthrodesis of the ankle has several advantages compared with open arthrodesis, including a smaller skin incision, less damage to the soft tissue around the joint, a lower risk of skin necrosis and infection, a lower incidence of postoperative infection and swelling, and better preservation of the contour of the surface of the joint, which maintains a larger contact area between the talus and tibia. We successfully performed arthroscopic arthrodesis of the tibiotalocalcaneal joints with intramedullary nails with fins in 9 ankles of 8 patients. Solid fusion was attained in all cases, except for 1 case of nonunion at the subtalar joint. We also corrected the alignment in 1 patient with a varus deformity. The fixation was strong, even in the case of poor bone quality, such as occurs in rheumatoid arthritis. The intramedullary nails with fins allowed for appropriate compression for bone consolidation without loss of rotational stability. Arthroscopic tibiotalocalcaneal arthrodesis, a less-invasive technique than conventional open surgery, is effective treatment, especially in patients with poor skin conditions secondary to diseases such as rheumatoid arthritis and diabetes mellitus. PMID- 21641831 TI - Hallux varus as complication of foot compartment syndrome. AB - Hallux varus can present as a congenital deformity or it can be acquired secondary to trauma, surgery, or neuromuscular disease. In the present report, we describe the presence of hallux varus as a sequela of calcaneal fracture with entrapment of the medial plantar nerve in the calcaneal tunnel and recommend that clinicians be wary of this when they clinically, and radiographically, evaluate patients after calcaneal fracture. PMID- 21641832 TI - Segment IV preserving middle hepatic vein retrieval in right lobe living donor liver transplantation. PMID- 21641833 TI - Lymph node counts as an indicator of quality at the hospital level in colorectal surgery. AB - BACKGROUND: Substantial evidence suggests that the number of lymph nodes examined in colorectal cancer (CRC) is a powerful predictor of outcomes. However, the lymph node count as a benchmark of quality in CRC is controversial. We sought to examine the impact of lymph node counts on disease-specific survival (DSS) of CRC patients at the hospital level. STUDY DESIGN: This study used data obtained between 1994 and 2003 from Region 5 of the California Cancer Registry. Hospitals in Region 5 of the California Cancer Registry were stratified according to the median number of nodes examined and grouped according to the median number of nodes examined, <7, 7 to 9, and >=10. These hospital groups were then evaluated for the frequency of meeting the 12-node threshold, frequency of positive lymph nodes, and DSS at the hospital level. RESULTS: Median number of nodes examined in group A was 4 (mean 5.6, SD 5.9), in group B was 8 (mean 9.7, SD 8.5), and in group C was 10 (mean 11.3, SD 9.2). In group A, 13.7%, in group B 32.8%, and in group C, 42.8% met the 12-node threshold. The frequency of N1 and N2 disease for group A was 20.7% and 9.1%, 19. 7% and 11.1% for group B, and 20.1% and 11.3% for group C (p = 0.12). Five-year DSS was 72.7% for group A, 73.7% for group B, and 76.7% for group C (p = 0.002). DSS survival of N0 patients for group A was 78.6%, 81.5% for group B, and 85.1% for group C (p < 0.0001). There was no statistically significant difference in DSS for N1 (p = 0.18) or N2 (p = 0.90) between the 3 groups. CONCLUSIONS: Lymph node counts can have value as a benchmark of surgical/pathologic quality in node-negative CRC. These results question the value of lymph node counts as a benchmark of surgical/pathologic quality for node positive CRC. PMID- 21641834 TI - Decisions, decisions: how program diversity influences residency program choice. AB - BACKGROUND: Recent studies suggest that students' feelings of fit with a residency program substantially influence students' ranking of the program. As diversity issues become increasingly focal concerns, we investigate how perception of gender and racial diversity of a program influences students' rankings of the program. We focus on students pursuing surgical specialties and ask whether diversity concerns are more prominent among applicants to surgical programs than among applicants to nonsurgical programs. STUDY DESIGN: We invited all interviewees at all residency programs at the Stanford University School of Medicine to participate in our study in the spring of 2009. Nineteen residency programs, amounting to 1,657 residency interviewees, participated. Sixty-eight percent (n = 1,132) responded to the survey. RESULTS: Women and under-represented minority applicants differ in their assessments from male and non-under represented minority applicants because women applying to surgical programs and under-represented minority students are less likely than others to perceive their prospective programs as diverse. However, perceived program diversity is an important factor that positively influences the program ranking decision for women and minorities pursuing surgical training. CONCLUSIONS: Surgical training programs that promote gender and racial diversity will likely be more successful in attracting women and minority students because women and minorities are especially sensitive to program diversity in both their perceptions and rankings of programs. Promoting women and minorities within programs and connecting women and minority applicants to outreach programs and mentors is pertinent to the recruitment of these traditionally under-represented groups to surgical programs. PMID- 21641835 TI - Modified hospital elder life program: effects on abdominal surgery patients. AB - BACKGROUND: Postsurgical functional decline is common in older patients and can lead to frailty and increased mortality. Comprehensive interventions such as the Hospital Elder Life Program (HELP) have been shown to be effective, but modifying the HELP to include only 3 key interventions might prove cost-effective for surgical patients. STUDY DESIGN: Consecutive patients from August 2007 through April 2009 (n = 179) were enrolled if they had undergone common elective abdominal surgical procedures, such as gastrectomy, cholecystectomy, and Whipple surgery. A modified HELP intervention consisting of early mobilization, nutritional assistance, and therapeutic (cognitive) activities implemented by a trained nurse was introduced on a surgical ward in May 2008. Patients enrolled before May 2008 received usual care and served as controls (n = 77). Those enrolled after the modified HELP intervention constituted the experimental group (n = 102). Changes in performance of activities of daily living, nutritional status, and cognitive function between admission and discharge were the primary end points. RESULTS: Independent of baseline functions, education, periampullary diagnosis, comorbidity, surgical procedure, and duration of surgery, patients in the HELP group declined significantly less on activities of daily living performance and nutritional status (p < 0.001) than controls. The delirium rate was also significantly lower in the HELP group (0%) than in the control group (16.7%) (p < 0.001). CONCLUSIONS: The modified HELP intervention effectively reduced older surgical patients' functional decline and delirium rates by hospital discharge. This program, conducted by a trained nurse, was not costly but did require commitment and ongoing cooperation between physician and nursing leadership to achieve compliance with the protocols. PMID- 21641836 TI - A modified DRIL procedure by flow reversal of an aborted distal cephalic vein for critical hand ischaemia. AB - Treatment of dialysis access-related hand ischaemia with preservation of the access remains an issue. We report the case of a patient presenting critical hand ischaemia 2 years after proximalisation of a distal radio-cephalic fistula with preservation of the original access. After valvulotomy, the distal cephalic vein was used as in situ bypass and directly anastomosed to the distal brachial artery, providing a direct flow to the hand. This procedure relieved the hand ischaemia without sacrificing the functioning fistula. Six months later, the fistula and bypass were still patent, showing that flow reversal of a previous fistula can be an efficient strategy to correct dialysis access-related hand ischaemia in selected cases. PMID- 21641837 TI - Diffuse dermal angiomatosis. AB - Diffuse dermal angiomatosis (DDA) is characterized clinically by painful erythematous lesions with ulcers and histologically by a benign, diffuse, and self-limited proliferation of tiny blood vessels in the superficial layers of the reticular dermis. Here we describe a case of DDA with leg ulcer. Erythematous lesions presented around the ulcer and angiogram revealed an occlusion of the superficial femoral artery. The erythematous lesions disappeared after revascularization. Although DDA is extremely rare, early correction of the ischemia in the peripheral artery should be taken into consideration. PMID- 21641838 TI - Circadian rhythms and cardiovascular health. AB - The functional organization of the cardiovascular system shows clear circadian rhythmicity. These and other circadian rhythms at all levels of organization are orchestrated by a central biological clock, the suprachiasmatic nuclei of the hypothalamus. Preservation of the normal circadian time structure from the level of the cardiomyocyte to the organ system appears to be essential for cardiovascular health and cardiovascular disease prevention. Myocardial ischemia, acute myocardial infarct, and sudden cardiac death are much greater in incidence than expected in the morning. Moreover, supraventricular and ventricular cardiac arrhythmias of various types show specific day-night patterns, with atrial arrhythmias--premature beats, tachycardias, atrial fibrillation, and flutter - generally being of higher frequency during the day than night--and ventricular fibrillation and ventricular premature beats more common, respectively, in the morning and during the daytime activity than sleep span. Furthermore, different circadian patterns of blood pressure are found in arterial hypertension, in relation to different cardiovascular morbidity and mortality risk. Such temporal patterns result from circadian periodicity in pathophysiological mechanisms that give rise to predictable-in-time differences in susceptibility-resistance to cyclic environmental stressors that trigger these clinical events. Circadian rhythms also may affect the pharmacokinetics and pharmacodynamics of cardiovascular and other medications. Knowledge of 24-h patterns in the risk of cardiac arrhythmias and cardiovascular disease morbidity and mortality plus circadian rhythm-dependencies of underlying pathophysiologic mechanisms suggests the requirement for preventive and therapeutic interventions is not the same throughout the day and night, and should be tailored accordingly to improve outcomes. PMID- 21641839 TI - Identification and validation of early biomarkers of osteoarthritis in companion animals: are we ready for the challenge? PMID- 21641840 TI - Calretinin immunostaining as an adjunct in the diagnosis of Hirschsprung disease. AB - Historically, the diagnosis of Hirschsprung disease was made by evaluating multiple hematoxylin and eosin-stained slides and performing acetylcholinesterase histochemical staining. Recently, calretinin immunohistochemical staining has been reported and found to be superior to acetylcholinesterase staining in the confirmation of aganglionosis. We retrieved tissue blocks from 23 patients with proven Hirschsprung disease from the archives of the Medical College of Georgia. In addition, we selected 23 control patients with ganglion cells. All cases were stained with calretinin, and the presence or absence of both intrinsic nerve fibers (INFs) and ganglion cells was scored by 4 pathologists with fairly strong agreement (kappa = 0.858). All cases of proven Hirschsprung disease were negative for INFs. Eighty-three percent of non-Hirschsprung patients were positive for INFs. Based on statistical analysis, the association between disease status and pathologist rating was statistically significant (P < .0001). We also found calretinin immunostaining to be a useful adjunctive modality in the diagnosis of Hirschsprung disease. PMID- 21641841 TI - Solid-pseudopapillary neoplasm of pancreas with long delayed liver metastasis. AB - Solid-pseudopapillary neoplasm (SPN) of the pancreas is a rare neoplasm that most commonly affects adolescent girls and young women. Solid-pseudopapillary neoplasm of the pancreas is considered to have malignant potential; 10% to 15% of cases are associated with metastasis, and these usually present at the time of initial diagnoses. Cases with metastases after resection are rare, and all reported cases have occurred less than 5 years after resection. We report a case of SPN in a 36 year-old woman who presented with liver metastasis 15.8 years after complete resection of the primary tumor in the pancreas. To the best of our knowledge, this is the longest time interval reported between resection of primary tumor and subsequent detection of metastatic disease. Solid-pseudopapillary neoplasm of the pancreas can present with metastases many years after resection of the primary tumor. Long-term follow-up is warranted, given the possibility of late metastasis. PMID- 21641842 TI - Remission of Cushing's disease with growth hormone replacement. AB - Treatment of Cushing's disease is often challenging. The success rates are not satisfactory and therapeutic interventions frequently causes hypopituitarism. We present three cases of remitting Cushing's disease complicated with growth hormone deficiency. Surprisingly, growth hormone replacement resulted in complete clinical and laboratory remission of Cushing's disease in these patients. However, one of the patients died of myocardial infarction. Effects of growth hormone on cortisol metabolism are discussed. PMID- 21641843 TI - Increased inspiratory and expiratory muscle strength following respiratory muscle strength training (RMST) in two patients with late-onset Pompe disease. AB - Respiratory muscle strength training (RMST) is an exercise-based intervention which targets respiratory muscle weakness. We implemented RMST in two patients with late-onset Pompe disease (LOPD), both who had received long-term enzyme replacement therapy and had severe respiratory weakness. Over 16-32 weeks, inspiratory muscle strength increased by 73-74%. Expiratory muscle strength increased 31-48% over 12-22 weeks. These findings suggest that RMST may increase respiratory muscle strength, even in the setting of LOPD and severe baseline weakness. PMID- 21641844 TI - Quality control for system count rate performance with scatter in gamma cameras. AB - We aimed to analyze the optimal conditions to carry out the periodical statistical control tests of the gamma camera count rate performance. First we focused in reproducing the actual R(20) value of the gamma camera response. Second we studied the variability of this parameter in the statistical control test. We performed a reference measurement, which consisted of the determination of the complete curve relating observed and incident count rates, the counting model describing it and the reference R(20). This reference determined the conditions for the statistical control tests and the way to analyze the results obtained. Results from three different gamma cameras were studied. Each gamma camera showed a different behavior and required specific data analysis. The optimal conditions to perform the statistical control test were determined in each case. Our procedure provides the information necessary to correlate the average value of R(20) obtained in the quality control test with the reference one. The critical requirement to perform any statistical control test, that is to have a reduced variability of the control variable, can be fulfilled in this case only for relatively high activities. PMID- 21641845 TI - [Toxic epidermal necrolysis with fatal ending due to the concurrent use of carbamazepine, cilostazol and omeprazol: a case report]. PMID- 21641846 TI - Developing a search engine for pharmacotherapeutic information that is not published in biomedical journals. AB - OBJECTIVE: To identify publishers of pharmacotherapeutic information not found in biomedical journals that focuses on evaluating and providing advice on medicines and to develop a search engine to access this information. METHODS: Compiling web sites that publish information on the rational use of medicines and have no commercial interests. Free-access web sites in Spanish, Galician, Catalan or English. Designing a search engine using the Google "custom search" application. RESULTS: Overall 159 internet addresses were compiled and were classified into 9 labels. We were able to recover the information from the selected sources using a search engine, which is called "AlquimiA" and available from http://www.elcomprimido.com/FARHSD/AlquimiA.htm. CONCLUSIONS: The main sources of pharmacotherapeutic information not published in biomedical journals were identified. The search engine is a useful tool for searching and accessing "grey literature" on the internet. PMID- 21641847 TI - Self-collected nasal swabs to detect infection and colonization: a useful tool for population-based epidemiological studies? AB - Population-based epidemiological studies on infectious diseases are limited by methodological problems that may not be encountered in other fields of epidemiology. The acute or asymptomatic nature of many infections hinders a timely diagnosis by trained personnel in a study centre, indicating the need for new collection methods of biological specimens. One alternative approach is to have the participants collect the specimens themselves, for instance nasal swabs for the detection of bacterial or viral pathogens. Although self-collection is widely accepted in clinical studies of specific populations (e.g., self collection of vaginal swabs by young women to diagnose sexually transmitted infections), it has not been employed much in population-based studies. Here, we review recent experience with self-collection of nasal swabs for the detection of microorganisms and discuss future prospects and applications for this technique. PMID- 21641848 TI - Subclinical signs in LRRK2 mutation carriers. AB - BACKGROUND: Several non-motor features have been reported to precede the motor signs of Parkinson's disease (PD) by several years. However, the time of onset of non-motor and motor symptoms is still debated. Healthy individuals carrying a PD related mutation are candidates for studying the earliest disease signs. OBJECTIVES: To describe clinically healthy family members of PD patients carrying a LRRK2 mutation (LRRK2-PD). METHODS: A total of 47 family members of LRRK2-PD patients were included in the present study and were screened for the p.G2019S and p.N1437H substitutions in the LRRK2 gene. A standardized case report form was filled out in each case, including general medical evaluation, neurological examination with UPDRS, an olfaction test, mood, sleep and cognitive questionnaires. RESULTS: Thirty-two study participants were positive, and 15 were negative for a LRRK2 mutation. Higher UPDRS motor scores, more frequent reports of urinary problems, and fewer hours of sleep were found in mutation carriers compared to non-carriers. The mutation carriers with UPDRS >=8 were all aged over 50 years, had shorter overall sleeping hours, more frequent urinary and constipation problems, higher mood scores and body mass index. Deterioration of olfaction was not detected in either group. CONCLUSION: Healthy LRRK2 mutation carriers presented subclinical parkinsonian motor and non-motor signs in the apparent absence of olfactory loss. Longitudinal studies will determine whether these changes precede alterations detectable by neuroimaging. PMID- 21641849 TI - Clinical prediction rules in the physiotherapy management of low back pain: a systematic review. AB - OBJECTIVE: To identify, appraise and determine the clinical readiness of diagnostic, prescriptive and prognostic Clinical Prediction Rules (CPRs) in the physiotherapy management of Low Back Pain (LBP). DATA SOURCES: MEDLINE, EMBASE, CINAHL, AMED and the Cochrane Database of Systematic Reviews were searched from 1990 to January 2010 using sensitive search strategies for identifying CPR and LBP studies. Citation tracking and hand-searching of relevant journals were used as supplemental strategies. STUDY SELECTION: Two independent reviewers used a two phase selection procedure to identify studies that explicitly aimed to develop one or more CPRs involving the physiotherapy management of LBP. Diagnostic, prescriptive and prognostic studies investigating CPRs at any stage of their development, derivation, validation, or impact-analysis, were considered for inclusion using a priori criteria. 7453 unique records were screened with 23 studies composing the final included sample. DATA EXTRACTION: Two reviewers independently extracted relevant data into evidence tables using a standardised instrument. DATA SYNTHESIS: Identified studies were qualitatively synthesized. No attempt was made to statistically pool the results of individual studies. The 23 scientifically admissible studies described the development of 25 unique CPRs, including 15 diagnostic, 7 prescriptive and 3 prognostic rules. The majority (65%) of studies described the initial derivation of one or more CPRs. No studies investigating the impact phase of rule development were identified. CONCLUSIONS: The current body of evidence does not enable confident direct clinical application of any of the identified CPRs. Further validation studies utilizing appropriate research designs and rigorous methodology are required to determine the performance and generalizability of the derived CPRs to other patient populations, clinicians and clinical settings. PMID- 21641850 TI - An exploration of familial associations in spinal posture defined using a clinical grouping method. AB - The primary aim of this study was to examine familial associations in spinal posture, defined using postural angles and a clinical classification method. A secondary aim was to investigate the reliability of clinical postural classification. Postural angles were calculated from sagittal photographs, while two experienced clinicians made use of standing sagittal images to classify participants into one of four postural groups (sway, flat, hyperlordotic, neutral). Parent-child associations in postural angles and postural groups were evaluated using Pearson's correlation and Fisher's exact test, respectively. Inter-rater reliability was expressed using percentage agreement and Kappa coefficients (K). Daughters whose father or mother had a hyperlordotic posture were 4.0 or 3.5 times, respectively, more likely to have a hyperlordotic posture than daughters whose parents did not have a hyperlordotic posture. These participants in the hyperlorotic group had a significantly higher body mass index than members of the other postural groups (p < 0.03). Percentage agreement between clinicians was 63.5% (K = 0.48). These results provide preliminary evidence of a familial association in the hyperlordotic posture and support the use of postural classification. PMID- 21641851 TI - CYP26B1 is a novel candidate gene for betel quid-related oral squamous cell carcinoma. AB - Substantial epidemiological data suggest a role for environmental factors (for example, the use of alcohol, betel quid (BQ), and cigarettes) in the occurrence of oral squamous cell carcinoma (OSCC), but the evidence for the genes involved has been inconsistent. This study was to investigate the role of CYP26B1, together with the use of alcohol, BQ, and cigarettes, on BQ-related OSCC. The association study (247 OSCC cases and 338 controls) was conducted to examine the possible interplay between CYP26B1 polymorphisms and alcohol, BQ, and cigarettes use. Additional gene expression was evaluated between OSCC tissue and adjacent normal tissue. The genetic polymorphism AA of CYP26B1 appeared to correlate with the risk of OSCC (OR=2.26; 95% CI, 1.35-3.80). Chewing BQ multiplicatively interacted with CYP26B1 AA to increase the OSCC risk (aOR=70.04; 95% CI, 13.62 360.11). The independent risk of OSCC was observed among BQ chewers with CYP26B1 AA, and compared with chewers with the CYP26B1 CC genotype (stratified aOR=2.88; 95% CI, 1.07-7.74). Increased expression of CYP26B1 was observed in tumor tissue compared with adjacent normal tissue. The CYP26B1 gene plays a novel role in the BQ dependent pathogenesis of OSCC. PMID- 21641852 TI - Management and outcome of head and neck squamous cell carcinomas in obese patients. AB - Head and neck squamous cell carcinomas are common lesions, related to chronic smoking and drinking behaviors. But in contrast to other cancers, effect of obesity on occurrence, diagnosis, treatment and prognosis of these tumors remains to date unknown. This is a retrospective review of 111 obese patients (sex ratio=6.4, median age=54.5 year old), treated between 1999 and 2007. Risk factors, tumoral localization and staging (41% stage I-II) were the same as in general population. However, we found 26.1% difficult pan-endoscopies, 54% ACE-27 comorbidity scores >=2 and 22.5% misstaged cervical lymphadenopathy. Treatment was based upon surgery (61%) or radiotherapy-chemotherapy (39%), and 37% of patients developed complications. Median follow up (38 months) and five-year overall survival (50%) are comparable to data in non obese patients. Although no direct relation between obesity and squamous cell carcinomas of the head and neck was found, obesity causes problems in tumor assessment and increases surgical complications rate. However, final good therapeutic tolerance and overall survival rate show that these patients should be managed like normal weighted ones. Receiving optimal treatments allow them to anticipate equivalent outcome as in general population. PMID- 21641853 TI - Synovial sarcoma involving skull base--a retrospective analysis of diagnosis and treatment of 21 cases in one institution. AB - To retrospectively review the diagnosis and treatment outcomes of 21 diagnosed with synovial sarcoma (SS) extending to skull base region and identify prognostic factors for local recurrence and distant metastasis. The clinical and pathological records of 21 synovial sarcoma cases involving skull base treated at Department of Oral and Maxillofacial Surgery, Affiliated to Ninth People's Hospital, Shanghai Jiao Tong University, from 2003 to 2008 were reviewed. Parameters including clinical, radiographic, histological findings, and treatment modalities were analyzed. Prognostic factors influencing overall survival, local recurrence, and metastasis were identified. Categorical variables were compared between groups for local recurrence and distant metastasis by using 2-tailed Fisher exact test. The sample consisted of 8 males and 13 females with a median age of 23. 42.9% of cases showed skull base bone erosion with 9.5% of cases presenting intracranial tumor extension. 52.4% of cases died from recurrence or metastasis with a median survival time of 11 months. Orbital involvement, perineural and cranial invasion along with other factors were statistically significant for their impact on overall survival. Local recurrence was associated with tumors located in the infratemporal space (p=0.01), perineural invasion (p=0.001), pathological grade 3 (p=0.005), and tumor size >5 cm (p=0.008). The aggressive behavior of skull base SS and its close proximity to vital structures make the diagnosis and management a challenge. The study reported high local recurrence rates despite surgical and postoperative radiotherapy. Adjuvant chemotherapy did not seem to affect distant metastasis rates. PMID- 21641854 TI - How do oomycete effectors interfere with plant life? AB - Oomycete genomes have yielded a large number of predicted effector proteins that collectively interfere with plant life in order to create a favourable environment for pathogen infection. Oomycetes secrete effectors that can be active in the host's extracellular environment, for example inhibiting host defence enzymes, or inside host cells where they can interfere with plant processes, in particular suppression of defence. Two classes of effectors are known to be host-translocated: the RXLRs and Crinklers. Many effectors show defence-suppressive activity that is important for pathogen virulence. A striking example is AVR3a of Phytophthora infestans that targets an ubiquitin ligase, the stabilisation of which may prevent host cell death. The quest for other effector targets and mechanisms is in full swing. PMID- 21641855 TI - Effects of environment on the photophysical characteristics of mesotetrakis methylpyridiniumyl porphyrin (TMPyP). AB - Porphyrins are an important class of organic molecules, with interesting linear and nonlinear optical properties given mainly by their extended pi-conjugation structure. Their photophysical properties can be greatly affected by the surrounding environment, which can be used to tune its final properties. Here we report on an experimental study of the photophysical properties of meso-tetrakis (methylpyridiniumyl) porphyrin (TMPyP) in aqueous and in several organic solvents and its interaction with micelles formed from negatively charged sodium dodecylsulphate (SDS), positively charged cetyl trimethyl ammonium bromide (CTAB) and neutral TRITON X-100. By using the Z-scan technique, flash-photolysis and time-resolved fluorescence techniques, we were able to evaluate the excited state dynamics of the TMPyP, and observed that the tetrapyrrole ring plays important role due to hydrogen bonds formation between nitrogen atom and water, while the side groups determine the porphyrin localization in non-aqueous micelle part. PMID- 21641856 TI - Investigation into the regiochemistry of some isoxazoles derived from 1,3-dipolar cycloaddition of 4-nitrobenzonitrile oxide with some dipolarophiles: a combined theoretical and experimental studies. AB - Reaction of 4-nitrobenzonitrile oxide (2) which was generated in situ with acrylo nitrile (3), methyl methacrylate (4) and allyl bromide (5) as dipolarphile afforded the new 7a, 8a and 9a compounds respectively. Reactivity and regiochemistry of these reactions were investigated using activation energy calculations and density functional theory (DFT)-based reactivity indexes. The theoretical 13C NMR chemical shifts of the cycloadducts which were obtained by GIAO method were comparable with the observed values. PMID- 21641857 TI - Synthesis, spectral and electrochemical studies of Cu(II) and Ni(II) complexes with new N2O2 ligands: a new precursor capable of depositing copper nanoparticles using thermal reduction. AB - Cu(II) and Ni(II) complexes of the general type [M(N2O2)] are described. The N2O2 ligands used are [N,N'-bis(2-hydroxy-6-methoxybenzylidene)propane-1,3-diamine] (HOMeSalpn) and [N,N'-bis(2-hydroxy-6-methoxybenzylidene)propane-1,2-diamine (HOMeSalpr). These complexes have been characterized by IR, UV-vis, CV, TG-DTA and 1H NMR spectroscopy. The electrochemical behavior of these complexes at a glassy carbon electrode in acetonitrile solution indicates that the first reduction process corresponding to Cu(II)-Cu(I) and Ni(II)-Ni(I) is electrochemically irreversible. The new copper complexes have been applied for the preparation of copper nanoparticles using non-ionic surfactant (Triton X-100) by thermal reduction. The copper nanoparticles with average size of 48nm were formed by thermal reduction of [N,N'-bis(2-hydroxy-6-methoxybenzylidene)propane 1,3-diamine]copper(II) in the presence of triphenylphosphine thus releasing the reduced copper and affording the high-purity copper nanoparticles. PMID- 21641858 TI - Synthesis, characterization, tautomerism and theoretical study of some new Schiff base derivatives. AB - New Schiff base derivatives were prepared by the condensation of 5-chloro and 5 bromo salicylaldehyde with bis(o-aminophenol)ethers. Five bis(o-nitrophenol)ether compounds were synthesized using some ditosylate, 1,3-dibromopropane and 1,4 dibromobuthane with o-nitrophenol. These compounds were reduced to bis(o aminophenol)ethers. The products have been characterized by elemental analysis, FTIR, 1H, 13C NMR, HETCOR and HMBC spectroscopic techniques. The tautomerisms of all of the Schiff bases compounds were determined in DMSO, CHCl3, C2H5OH and C6H12 solvents and in both acidic and basic media using the UV-vis spectrophotometric method. The heat of formation (DeltaHf), enthalpy (DeltaH), entropy (DeltaS), Gibbs free energy (DeltaGf and DeltaG), stable isomers, conformations and tautomers of the synthesized compounds are calculated using the MOPAC2009 (PM6) program. PMID- 21641859 TI - Serial electrophysiological changes in uraemic patients with tetrodotoxin intoxication. PMID- 21641860 TI - A comparative study of cortical responses evoked by transcutaneous electrical vs CO(2) laser stimulation. AB - OBJECTIVES: A novel non-invasive method for nociceptive electrical stimulation of the skin has been recently introduced by using a planar concentric stimulating electrode (CE). We compared the cortical potentials induced by a CE vs laser stimulator in healthy subjects using a multichannel recording. METHODS: Cortical potentials were recorded in 11 healthy subjects by 54 scalp electrodes, stimulating the skin of the right hand and the right supra-orbital zone by the CE and laser stimulator settled two levels above the individual pain threshold. RESULTS: The latency difference between N1, N2, and P2 evoked by the CE vs laser stimulator was larger than the receptor activation time of 40 ms and larger following stimulation of the upper limbs than of the head. The amplitudes and topographic distribution of the cortical waves did not differ between the two stimulation types. CONCLUSIONS: A-beta fibre co-activation may be induced by CE electrodes, as suggested by latency gaps. Nevertheless, CE-evoked potentials showed similarity in amplitude, morphology and topographic representation with laser-induced ones. SIGNIFICANCE: At present, CE-evoked potentials cannot be considered a reliable measure of nociceptive pathway function. PMID- 21641861 TI - Atypical EEG complexity in autism spectrum conditions: a multiscale entropy analysis. AB - OBJECTIVE: Intrinsic complexity subserves adaptability in biological systems. One recently developed measure of intrinsic complexity of biological systems is multiscale entropy (MSE). Autism spectrum conditions (ASC) have been described in terms of reduced adaptability at a behavioural level and by patterns of atypical connectivity at a neural level. Based on these observations we aimed to test the hypothesis that adults with ASC would show atypical intrinsic complexity of brain activity as indexed by MSE analysis of electroencephalographic (EEG) activity. METHODS: We used MSE to assess the complexity of EEG data recorded from 15 participants with ASC and 15 typical controls, during a face and chair matching task. RESULTS: Results demonstrate a reduction of EEG signal complexity in the ASC group, compared to typical controls, over temporo-parietal and occipital regions. No significant differences in EEG power spectra were observed between groups, indicating that changes in complexity values are not a reflection of changes in EEG power spectra. CONCLUSIONS: The results are consistent with a model of atypical neural integrative capacity in people with ASC. SIGNIFICANCE: Results suggest that EEG complexity, as indexed by MSE measures, may also be a marker for disturbances in task-specific processing of information in people with autism. PMID- 21641862 TI - Is neurophysiology a valuable marker of pre-clinical stages of Lewy Body Disorders? PMID- 21641863 TI - Transcranial magnetic stimulation reveals high test-retest reliability for phosphenes but not for suppression of visual perception. AB - OBJECTIVE: To investigate test-retest reliability of visual cortical excitatory and inhibitory phenomena. METHODS: Transcranial magnetic stimulation (TMS) was applied over occipital cortex twice in 22 healthy young adults with at least a one-month interval between both measurements. The test-retest reliability of the phosphenes and TMS-induced suppression of visual perception was assessed using correlation and calculation of the repeatability coefficient. RESULTS: Both analyses revealed a high reliability for phosphenes but not for the suppression of visual perception. CONCLUSIONS: It seems likely that the phosphenes may be better used than the TMS-induced suppression of visual perception in experiments which need repeated measurements (e.g., longitudinal studies or studies with pharmacological and non-pharmacological interventions). SIGNIFICANCE: The study demonstrates a rather limited value of the TMS-induced suppression of visual perception for studies with repeated measurements. PMID- 21641864 TI - Analyses of peptides in sake mash: forming a profile of bitter-tasting peptides. AB - Some oligopeptides and amino acids have a strong influence on the sensory qualities of sake, but the formation process of such compounds in sake mash has not been well elucidated. In this study, we investigated the formation process of bitter-tasting peptides derived from rice proteins in sake mash, because knowledge about their formation may contribute to the quality control of sake. We analyzed rice protein hydrolysates in sake mash, as well as in the enzymatic digest of steamed rice grains digested by either sake-koji or by crude enzyme extracted from sake-koji. SDS-PAGE showed that a smaller amount of polypeptides (>M.W. 10,000) accumulated in the supernatant of sake mash than in either enzymatic digest. The concentration of peptides in the supernatant of sake mash increased gradually from the early stages of fermentation. Five bitter-tasting peptides (No. 9, =75 years of age. METHODS: In a prospective population-based study, 534 patients (23% >=75 years) were consecutively included before surgery. HRQOL, medical and socio-demographic variables were measured by questionnaires at baseline, 6 and 12 months after surgery. HRQOL was measured by the short-form health survey (SF-36). RESULTS: Five hundred and twenty patients were alive 12 months after surgery, 89% responded after 6 and 12 months. Older patients as well as younger patients had a clear overall improvement in HRQOL over the first year after cardiac surgery, more specifically during the first 6 months. The same pattern was found in self-reported NYHA class which improved from baseline to 6 months and stabilized between 6 and 12 months. The only SF-36 scale with different development was Role Physical where younger patients improved more than older patients. Before surgery, patients had substantially lower scores than the population norms. However, on most dimensions of HRQOL older patients reached the level of the norm population after surgery. CONCLUSION: A selected group of elderly patients can undergo cardiac surgery with excellent results concerning survival and HRQOL. This is of major importance both discussing health care resources and decision making concerning individual patients. PMID- 21641871 TI - Antenatal diagnosis of bladder/cloacal exstrophy: challenges and possible solutions. AB - OBJECTIVE: To identify the pitfalls in accurate antenatal diagnosis of bladder exstrophy (BE) and cloacal exstrophy (CE), and thus understand the challenges for antenatal counselling. METHODS: A prospectively maintained bladder exstrophy database of antenatal and live born referrals for BE/CE was used to identify patients. Data were collected about the antenatal scan findings and the outcome of pregnancy. RESULTS: Between 2003 and 2009, 40 new babies with BE/CE were referred and of them 10 had an antenatal diagnosis. Five patients did not have a diagnosis despite suspicious findings noted on antenatal scans and another three had a wrong diagnosis of BE/CE. Of the 16 referrals with antenatal suspicion of BE/CE, 5 opted for termination. At the 20-week scan, it was possible to identify the gender of the fetus in 3/16 cases only. CONCLUSIONS: Only a quarter of the babies born with BE/CE had received an antenatal diagnosis. Raising awareness about the condition amongst radiographers, and facilitating further scanning by a specialist fetal management unit if suspicious findings are noted, is crucial for improving the rate of detection. An antenatal diagnosis may not be reliable, and difficulty in identifying gender at the 20-week scan adds to the complexity of antenatal counselling. Magnetic resonance imaging and karyotyping may provide additional helpful information. PMID- 21641867 TI - Intermittent versus continuous oxaliplatin and fluoropyrimidine combination chemotherapy for first-line treatment of advanced colorectal cancer: results of the randomised phase 3 MRC COIN trial. AB - BACKGROUND: When cure is impossible, cancer treatment should focus on both length and quality of life. Maximisation of time without toxic effects could be one effective strategy to achieve both of these goals. The COIN trial assessed preplanned treatment holidays in advanced colorectal cancer to achieve this aim. METHODS: COIN was a randomised controlled trial in patients with previously untreated advanced colorectal cancer. Patients received either continuous oxaliplatin and fluoropyrimidine combination (arm A), continuous chemotherapy plus cetuximab (arm B), or intermittent (arm C) chemotherapy. In arms A and B, treatment continued until development of progressive disease, cumulative toxic effects, or the patient chose to stop. In arm C, patients who had not progressed at their 12-week scan started a chemotherapy-free interval until evidence of disease progression, when the same treatment was restarted. Randomisation was done centrally (via telephone) by the MRC Clinical Trials Unit using minimisation. Treatment allocation was not masked. The comparison of arms A and B is described in a companion paper. Here, we compare arms A and C, with the primary objective of establishing whether overall survival on intermittent therapy was non-inferior to that on continuous therapy, with a predefined non inferiority boundary of 1.162. Intention-to-treat (ITT) and per-protocol analyses were done. This trial is registered, ISRCTN27286448. FINDINGS: 1630 patients were randomly assigned to treatment groups (815 to continuous and 815 to intermittent therapy). Median survival in the ITT population (n=815 in both groups) was 15.8 months (IQR 9.4-26.1) in arm A and 14.4 months (8.0-24.7) in arm C (hazard ratio [HR] 1.084, 80% CI 1.008-1.165). In the per-protocol population (arm A, n=467; arm C, n=511), median survival was 19.6 months (13.0-28.1) in arm A and 18.0 months (12.1-29.3) in arm C (HR 1.087, 0.986-1.198). The upper limits of CIs for HRs in both analyses were greater than the predefined non-inferiority boundary. Preplanned subgroup analyses in the per-protocol population showed that a raised baseline platelet count, defined as 400,000 per MUL or higher (271 [28%] of 978 patients), was associated with poor survival with intermittent chemotherapy: the HR for comparison of arm C and arm A in patients with a normal platelet count was 0.96 (95% CI 0.80-1.15, p=0.66), versus 1.54 (1.17-2.03, p=0.0018) in patients with a raised platelet count (p=0.0027 for interaction). In the per-protocol population, more patients on continuous than on intermittent treatment had grade 3 or worse haematological toxic effects (72 [15%] vs 60 [12%]), whereas nausea and vomiting were more common on intermittent treatment (11 [2%] vs 43 [8%]). Grade 3 or worse peripheral neuropathy (126 [27%] vs 25 [5%]) and hand-foot syndrome (21 [4%] vs 15 [3%]) were more frequent on continuous than on intermittent treatment. INTERPRETATION: Although this trial did not show non inferiority of intermittent compared with continuous chemotherapy for advanced colorectal cancer in terms of overall survival, chemotherapy-free intervals remain a treatment option for some patients with advanced colorectal cancer, offering reduced time on chemotherapy, reduced cumulative toxic effects, and improved quality of life. Subgroup analyses suggest that patients with normal baseline platelet counts could gain the benefits of intermittent chemotherapy without detriment in survival, whereas those with raised baseline platelet counts have impaired survival and quality of life with intermittent chemotherapy and should not receive a treatment break. FUNDING: Cancer Research UK. PMID- 21641872 TI - Robotic-assisted laparoscopic extravesical ureteral reimplantation: an initial experience. AB - OBJECTIVE: There are many emerging techniques using robotic-assisted laparoscopy (RAL) in pediatrics. We performed a retrospective review of our first patients who underwent RAL extravesical ureteral reimplantation. MATERIALS/METHODS: Between October 2007 and May 2010, a single surgeon performed RAL extravesical ureteral reimplantation in 17 patients. Six patients underwent bilateral reimplantation, resulting in a total of 23 ureters repaired. There were 16 females and 1 male (mean age 6.23 years). Four patients had prior Deflux injection. Postoperative reflux status was assessed by voiding cystourethrogram. RESULTS: 16 patients (22 ureters) were compliant with follow up. Mean follow up was 11.5 months. Mean anesthetic time was 3 h, 57 min for unilateral and 4 h, 45 min for bilateral repair. Complete vesicoureteral reflux resolution was seen in 20 ureters (90.9%), downgrading in one ureter, and unchanged persistent reflux in one ureter. Average hospital stay was 1.3 days. No patients required postoperative catheterization at discharge. CONCLUSIONS: Outcomes for new procedures can be variable and unpredictable as the technique evolves. Given the high success rates of open reimplantation, a minimally invasive technique must show comparable results if it is to play a continuing role. Our initial results are encouraging, but prospective analyses are required to outline the future role of RAL ureteral reimplantation. PMID- 21641873 TI - Intradural hemangiopericytoma of the thoracic spine: a case report. AB - BACKGROUND CONTEXT: Hemangiopericytoma (HPC) occurs infrequently in the central nervous system. Spinal involvement is particularly uncommon; and intradural localization is rare. Here, we describe an intradural extramedullary thoracic HPC that went undiagnosed initially on computed tomography scan of the abdomen. PURPOSE: To describe the clinical presentation and operative management of a patient diagnosed with an intradural extramedullary thoracic HPC that was missed on initial workup. We also describe the pathologic features of HPC. STUDY DESIGN: Case report. METHODS: Chart review and literature search. CASE: A 58-year-old man presented with acute weakness of the lower extremities and bladder and bowel incontinence. Magnetic resonance imaging of his spine revealed a T10 intradural extramedullary lesion that displaced the cord to the right. RESULTS: The patient was taken emergently to surgery for T9-T11 laminectomy and en bloc resection of the tumor. The lesion was identified and resected. Histology revealed randomly oriented tumor cells with irregular capillaries consistent with HPC. Postoperatively, the patient had an improved neurological examination, and he continued to do so with intense physical therapy. CONCLUSION: The standard treatment for HPC is surgery when the lesion is resectable. Despite gross total resection, there is still a high risk of recurrence and metastasis; therefore, patients should be followed up closely by their physicians with serial postoperative clinical examinations and radiographic imaging. PMID- 21641874 TI - Cost-effectiveness of multilevel hemilaminectomy for lumbar stenosis-associated radiculopathy. AB - BACKGROUND CONTEXT: Laminectomy for lumbar stenosis-associated radiculopathy is associated with improvement in pain, disability, and quality of life. However, given rising health-care costs, attention has been turned to question the cost effectiveness of lumbar decompressive procedures. The cost-effectiveness of multilevel hemilaminectomy for radiculopathy remains unclear. PURPOSE: To assess the comprehensive medical and societal costs of multilevel hemilaminectomy at our institution and determine its cost-effectiveness in the treatment of degenerative lumbar stenosis. STUDY DESIGN: Prospective single cohort study. PATIENT SAMPLE: Fifty-four consecutive patients undergoing multilevel hemilaminectomy for lumbar stenosis-associated radiculopathy after at least 6 months of failed conservative therapy were included. OUTCOME MEASURES: Self-reported measures were assessed using an outcomes questionnaire that incorporated total back-related medical resource utilization, missed work, and improvement in leg pain (visual analog scale for leg pain [VAS-LP]), disability (Oswestry Disability Index [ODI]), quality of life (Short Form-12 [SF-12]), and health state values (quality adjusted life years [QALYs], calculated from EuroQuol 5D [EQ-5D] with US valuation). METHODS: Over a 2-year period, total back-related medical resource utilization, missed work, and improvement in leg pain (VAS-LP), disability (ODI), quality of life (SF-12), and health state values (QALYs, calculated from EQ-5D with US valuation) were assessed. Two-year resource use was multiplied by unit costs based on Medicare national allowable payment amounts (direct cost), and patient and caregiver workday losses were multiplied by the self-reported gross of-tax wage rate (indirect cost). Mean total 2-year cost per QALY gained after multilevel hemilaminectomy was assessed. RESULTS: Compared with preoperative health states reported after at least 6 months of medical management, a significant improvement in VAS-LP, ODI, and SF-12 (physical and mental components) was observed 2 years after multilevel hemilaminectomy, with a mean 2 year gain of 0.72 QALYs. Mean+/-standard deviation total 2-year cost of multilevel hemilaminectomy was $24,264+/-10,319 (surgery cost, $10,220+/-80.57; outpatient resource utilization cost, $3,592+/-3,243; and indirect cost, $10,452+/-9,364). Multilevel hemilaminectomy was associated with a mean 2-year cost per QALY gained of $33,700. CONCLUSIONS: Multilevel hemilaminectomy improved pain, disability, and quality of life in patients with lumbar stenosis-associated radiculopathy. Total cost per QALY gained for multilevel hemilaminectomy was $33,700 when evaluated 2 years after surgery with Medicare fees, suggesting that multilevel hemilaminectomy is a cost-effective treatment of lumbar radiculopathy. PMID- 21641875 TI - Techniques of lumbar-sacral spine fusion in spondylosis: systematic literature review and meta-analysis of randomized clinical trials. AB - BACKGROUND CONTEXT: Spine fusions can be performed through different techniques and are used to treat a number of vertebral pathologies. However, there seems to be no consensus regarding which technique of fusion is best suited to treat each distinct spinal disease or group of diseases. PURPOSE: To study the effectiveness and complications of the different techniques used for spinal fusion in patients with lumbar spondylosis. STUDY DESIGN: Systematic literature review and meta analysis. SAMPLE: Randomized clinical studies comparing the most commonly performed surgical techniques for spine fusion in lumbar-sacral spondylosis, as well as those reporting patient outcome were selected. OUTCOME MEASURES: Identify which technique, if any, presents the best clinical, functional, and radiographic outcome. METHODS: Systematic literature review and meta-analysis based on scientific articles published and indexed to the following databases: PubMed (1966-2009), Cochrane Collaboration-CENTRAL, EMBASE (1980-2009), and LILACS (1982 2009). The general search strategy focused on the surgical treatment of patients with lumbar-sacral spondylosis. RESULTS: Eight studies met the inclusion criteria and were selected with a total of 1,136 patients. Meta-analysis showed that patients who underwent interbody fusion presented a significantly smaller blood loss (p=.001) and a greater rate of bone fusion (p=.02). Patients submitted to fusion using the posterolateral approach had a significantly shorter operative time (p=.007) and less perioperative complications (p=.03). No statistically significant difference was found for the other studied variables (pain, functional impairment, and return to work). CONCLUSIONS: The most commonly used techniques for lumbar spine fusion in patients with spondylosis were interbody fusion and posterolateral approach. Both techniques were comparable in final outcome, but the former presented better rates of fusion and the latter the less complications. PMID- 21641876 TI - Gender differences in cardiovascular risk: a varying variable. PMID- 21641877 TI - Recruitment pattern of sympathetic neurons during breath-holding at different lung volumes in apnea divers and controls. AB - We tested the hypothesis that breath-hold divers (BHD) attain higher level of sympathetic activation than controls due to the duration of breath-hold rather than a different recruitment strategy. In 6 control subjects and 8 BHD we measured muscle sympathetic neural activity (MSNA) prior to and during functional residual capacity (FRC) and total lung capacity (TLC) breath-holding. On a subset of subjects we applied a new technique for the detection of action potentials (APs) in multiunit MSNA. Compared with controls, BHD group had lower burst AP content (13+/-7 vs. 6+/-3AP/burst; P=0.05) and number of active clusters (5+/-1 vs. 3+/-1clusters/burst; P=0.05) at baseline. However, the overall sympathetic AP/unit-time was comparable between the groups (131+/-105 vs. 173+/-152AP/min; P=0.62) due to increased burst frequency in BHD group (20+/-4bursts/min) vs. controls (13+/-3bursts/min) (P=0.039). The achieved level in total MSNA during FRC breath-holds was higher in divers (2298+/-780 vs. 1484+/-575a.u./min; P=0.039). Total MSNA at the end of TLC breath-hold was comparable between the groups (157+/-50 (controls) vs. 214+/-41s (BHD); P=0.61). FRC and TLC breath holds increased AP frequency, burst AP content and active clusters/bursts in both groups but the response magnitude was determined by the type of the breath-hold. The divers used fewer number of APs/burst and active clusters/burst. In both groups breath-holds resulted in similar increases in MSNA which were reached both by an increase in firing frequency and by recruitment of previously silent, larger (faster conducting) sympathetic neurons, and possibly by repeated firing within the same burst. PMID- 21641878 TI - Development and validation of a sensitive liquid chromatography/tandem mass spectrometry method for the determination of exemestane in human plasma. AB - Exemestane, irreversible steroidal aromatase inhibitor, acts as a false substrate for aromatase enzyme and significantly lowers circulating estrogen concentrations in postmenopausal women with hormone-sensitive breast cancer. A sensitive bioanalytical method was developed and validated to study pharmacokinetics of exemestane. The method was based on liquid-liquid extraction of exemestane with methyl t-butyl ether followed by reversed-phase liquid chromatography. Positive electrospray ionization tandem mass spectrometry in multiple reaction monitoring mode was applied for detection of exemestane. Anastrozole was used as internal standard. Calibration curve, fitted to 1/x2 weighted linear regression model, was linear in the range of 0.1-40.0 ng/mL. Intra-run precision and accuracy were 1.80 3.17% and 103.4-111.5%, respectively. Inter-run precision and accuracy measured within 3 days were 3.37-4.19% and 101.8-109.6%, respectively. Extraction recoveries of exemestane and internal standard were 79.7-86.2% and 82.9-83.6%, respectively. The method was fully validated and may be applied to pharmacokinetic studies in humans after a single dose administration of 25mg exemestane tablets. PMID- 21641879 TI - Isolation and identification of antiplasmodial N-alkylamides from Spilanthes acmella flowers using centrifugal partition chromatography and ESI-IT-TOF-MS. AB - The development of new antiplasmodial drugs is of primary importance due to the growing problem of multi-drug resistance of malaria parasites. Spilanthes acmella, a plant traditionally used for the treatment of toothache, was targeted as a lead for its potential antiplasmodial activity. A systematic approach for investigating a suitable centrifugal partition chromatography (CPC) solvent system for N-alkylamides separation was reported. The partition behavior of three N-alkylamides has been studied using several biphasic solvent mixtures in search of an adequate CPC solvent system for this class of compounds. Major N alkylamides in S. acmella were isolated from a methanolic crude extract of flowers by CPC with the solvent system heptanes-ethyl acetate-methanol-water (3:2:3:2, v/v/v/v). Four N-alkylamides were purified and the structures were illustrated by electrospray ionization-ion trap-time of flight-mass spectrometry (ESI-IT-TOF-MS), 1H nuclear magnetic resonance (1H NMR) and 13C nuclear magnetic resonance (13C NMR). The CPC fractions, which contained natural mixtures of phytochemicals, demonstrated significantly higher antiplasmodial activity compared to corresponding purified N-alkylamides, thus suggesting that interactions between these N-alkylamides may potentiate antiplasmodial bioactivity. PMID- 21641880 TI - Assay for determination of daunorubicin in cancer cells with multidrug resistance phenotype. AB - A sensitive assay for direct determination of intracellular level of daunorubicin (DRN) in resistant leukemia cells with overexpressed P-glycoprotein has been developed. This assay is based on a rapid separation of cells from media and fast cut-off of DRN transportation by centrifugation of cells through a layer of silicone oil. Cell pellets were extracted using 1% (v/v) formic acid in 50% (v/v) ethanol in water. The cell extracts were subsequently analysed by liquid chromatography (HPLC) coupled a low-energy collision tandem mass spectrometer equipped with an electrospray ionization source (ESI-CID-MS/MS) operated in the multiple-reaction monitoring (MRM) mode. Calibration curve was linear from 0.4 to 250nM with correlation coefficient (r2) better than 0.998. The limit of quantitation (LOQ) was 0.4 nM. The assay has been successfully applied to a determination of intracellular content of daunorubicin in sensitive K562 and resistant K562/Dox and K562/HHT300 cells. PMID- 21641881 TI - Three phases hollow fiber LPME combined with HPLC-UV for extraction, preconcentration and determination of valerenic acid in Valeriana officinalis. AB - In the present work, the applicability of hollow fiber-based liquid phase microextraction (HF-LPME) was evaluated for the extraction and preconcentration of valerenic acid prior to its determination by reversed-phase HPLC/UV. The target drug was extracted from 5.0 mL of aqueous solution with pH 3.5 into an organic extracting solvent (dihexyl ether) impregnated in the pores of a hollow fiber and finally back extracted into 10 MU L of aqueous solution with pH 9.5 located inside the lumen of the hollow fiber. In order to obtain high extraction efficiency, the parameters affecting the HF-LPME, including pH of the donor and acceptor phases, type of organic phase, ionic strength, the volume ratio of donor to acceptor phase, stirring rate and extraction time were studied and optimized. Under the optimized conditions, enrichment factor up to 446 was achieved and the relative standard deviation (RSD) of the method was 4.36% (n = 9). The linear range was 7.5-850 MUg L-1 with correlation coefficient (r2=0.999), detection limits was 2.5 MUg L-1 and the LOQ was 7.5 MUg L-1. The proposed method was evaluated by extraction and determination of valerenic acid in some Iranian wild species of Valerianaceae. PMID- 21641882 TI - Studies of genomic copy number changes in human cancers reveal signatures of DNA replication stress. AB - Human cancers are characterized by the presence of genomic instability. Recently, two studies have catalogued the presence of a specific class of genomic aberrations, large deletions and insertions, in a few thousand human cancers and reported that most of the prevalent recurrent focal deletions targeted common fragile sites and large genes. In various experimental systems, deletions in common fragile sites and large genes have been linked to the presence of DNA replication stress. Thus, taken together, these results suggest the presence of DNA replication stress in human cancers, consistent with the recently proposed oncogene-induced DNA damage model for cancer development. PMID- 21641883 TI - [The value of 18FDG-PET for localization of ectopic Cushing's syndrome due to occult bronchial carcinoid]. PMID- 21641884 TI - [Acute hallucinatory psychosis secondary to oral glucocorticoid treatment in a patient diagnosed with Sheehan's syndrome]. PMID- 21641885 TI - Testosterone in female. PMID- 21641886 TI - Persistently positive gliadin antibodies without transglutaminase antibodies in the elderly: gluten intolerance beyond coeliac disease. AB - BACKGROUND: The specificity of the conventional gliadin antibody test is considered low. AIMS: We explored whether gliadin antibody(AGA)-positivity without tissue transglutaminase antibodies (tTGA) is persistent in the elderly population and whether such positivity indicates overt or potential coeliac disease in genetically predisposed individuals. METHODS: AGA and tissue transglutaminase antibody were measured in 2089 elderly individuals twice with a three-year interval. AGA-positive but tissue transglutaminase antibody-negative subjects with coeliac-type human leucocyte antigen (HLA) were examined and underwent gastroduodenal endoscopy (cases). Small-bowel mucosal villous morphology and densities of CD3+ and gammadelta+ intraepithelial lymphocytes and the occurrence of tissue transglutaminase-specific IgA deposits were analysed. Randomly selected persistently AGA-negative age- and sex-matched subjects served as controls. RESULTS: AGA-positivity was persistent in 81% of those initially positive. Amongst the 49 clinically studied and 36 endoscopied cases only one (2.8%) had coeliac disease. Many (54%) showed signs of inflammation in the biopsy, without villous atrophy. Coeliac-type HLA was not over-represented in the persistently AGA-positive compared to the general population. Persistently AGA positive coeliac-type HLA-positive subjects had more gastrointestinal symptoms than AGA-negative controls. CONCLUSIONS: AGA-positivity is often persistent. Overt coeliac disease is seldom found behind persistent AGA-positivity, but this characteristic is associated with mucosal inflammation and gastrointestinal symptoms at least in HLA-positive individuals. PMID- 21641887 TI - Misleading aspect of a metastatic melanoma detected by wireless capsule endoscopy. PMID- 21641888 TI - Relapse factors for ileal neuroendocrine tumours after curative surgery: a retrospective French multicentre study. AB - AIM: To evaluate the characteristics of postoperative relapse, predictive factors and time to relapse after curative surgery for well-differentiated neuroendocrine tumours of the ileum, without hepatic or other distant metastases. METHODS: Clinical data of patients entered into the Groupe d'etude des Tumeurs Endocrines database were collected and analysed retrospectively to identify factors predictive of relapse. RESULTS: Among 100 patients followed for a median of 56.5 (range 1-290) months, 42 relapsed after a median follow-up of 57.5 (range 6-176) months, with liver lesions in 27 (64.3%). Median disease-free survival (Kaplan Meier) was 88 months (95% confidence interval 72-115). Disease-free survival was shorter for emergency surgery patients (p<0.01), patients with distant mesenteric lymph-node metastases (p<0.01), with fortuitous diagnosis (p=0.02), with tumour diameter >20mm (p=0.02), and those with multiple tumours (p=0.07). Multivariate analysis retained emergency surgery (odds-ratio 4.04 [95% confidence interval 2.01-8.11]), distant mesenteric lymph-node metastases (odds-ratio 2.53 [95% confidence interval 1.22-5.25]), and multiple tumours (odds-ratio 2.14 [95% confidence interval 1.01-4.50]), as being significantly associated with relapse. CONCLUSION: Patients who underwent emergency surgery, with distant mesenteric lymph-node metastases or with multiple ileal tumours relapsed earlier. Closer monitoring for the patients with these risk factors may be required. PMID- 21641889 TI - Fatal gastric bleeding during sorafenib treatment for hepatocellular carcinoma recurrence after liver transplantation. PMID- 21641890 TI - Postembolization small bowel ischaemia. PMID- 21641891 TI - Conservative management of liver abscess complicated by hepatogastric fistula. PMID- 21641892 TI - Predicting consideration of cosmetic surgery in a college population: a continuum of body image disturbance and the importance of coping strategies. AB - Elective cosmetic surgeries are increasing in the American population with reasons linked to body image disturbance and body dysmorphic disorder (BDD). Little research exists documenting the continuum of body image disturbance and its relationship to seeking surgery. The present research examined data from 544 participants, 55 of whom were diagnosable with BDD. Using assessments for body image disturbance, problematic coping strategies, and BDD symptomatology, results provided evidence for a continuum of body image distress. Logistic regression analysis supported the hypothesis that increased levels of body image disturbance and one type of problematic coping strategy (Appearance Fixing) predicted consideration of cosmetic surgery. Of participants diagnosable with BDD, those who considered cosmetic surgery showed more severe body image disturbance and problematic coping than those who did not consider surgery. These results have implications for pre-surgical assessment as well as psychological interventions rather than invasive medical interventions. PMID- 21641893 TI - Marked for life? A prospective study of tattoos on appearance anxiety and dissatisfaction, perceptions of uniqueness, and self-esteem. AB - Previous studies on psychosocial aspects of tattooing have not examined prospective changes in self- and body-related attitudes as a result of obtaining a tattoo. In the present study, 82 British residents obtaining their first tattoo completed measures of state appearance anxiety and dissatisfaction prior to, and immediately after, obtaining a tattoo. They also completed measures of trait body appreciation, distinctive appearance investment, self-ascribed uniqueness, social physique anxiety, and self-esteem before obtaining a tattoo and three weeks later. Results showed that both women and men had significantly lower appearance anxiety and dissatisfaction immediately after obtaining their tattoo, and significantly higher body appreciation, distinctive appearance investment, self ascribed uniqueness, and self-esteem after three weeks. Women reported greater social physique anxiety after three weeks, whereas men reported lower anxiety. These results are discussed in relation to the positive impacts of obtaining body art and the mainstreaming of tattooing in Western societies. PMID- 21641894 TI - Modification of McGregor fan flap for reconstruction of full thickness defect of lower lip and vermilion. PMID- 21641895 TI - Case report: improving the speed and accuracy of melanoma sentinel node biopsy with 3D intra-operative imaging. AB - This case reports our first experience of using real-time intra-operative 3D imaging for sentinel lymph node biopsy in melanoma. The 3D imaging enabled ready identification and, excision of the sentinel node, by pinpointing its exact location. PMID- 21641896 TI - A fibrous band associated with the non-coronary aortic valve cusp in a dog. AB - A fibrous band connecting the middle of the free edge (nodulus Arantii) of the non-coronary aortic valve cusp to the ascending aorta just above the level of the non-coronary sinus of Valsalva was observed in an asymptomatic, 11-year-old, male Border Collie. The fibrous band was unrelated to the cause of the death in this dog. Such fibrous bands are usually reported in humans with congenital bicuspid aortic valves. To our knowledge, this is the first report of a fibrous band in the aortic valve in a domestic animal. PMID- 21641897 TI - Gerbode defect associated with blunt trauma in a dog. AB - A 6 year-old Labrador retriever was presented after being struck by a car. A ventricular arrhythmia, attributed to myocardial trauma, developed 12 h post trauma. Echocardiography revealed lesions consistent with a subaortic paramembranous ventricular septal defect (VSD) with shunting of blood from the left ventricle to the right atrium (Gerbode defect). A right-to-left shunting atrial septal defect (ASD) was visualised. Pleural and peritoneal effusions developed within 48 h. Fifteen days post-trauma flow across the ASD was left-to right while left-to-right shunting across the VSD persisted. No cavitary effusions were detected at 15 days post-trauma or subsequently. PMID- 21641898 TI - Reversed anatomic distal femur locking plate for periprosthetic hip fracture fixation. AB - The incidence of femoral fracture on hip prosthesis is increasing. Plate fixation is the method of choice when the prosthesis is stable. In fracture with proximal extension, the quality of the bone fixation is critical and, despite the development of anatomic plates, may be endangered when there are too few proximal screws. To resolve this issue, we recommend using a reversed LCPTM anatomic distal femoral Less Invasive Stabilization System (LISSTM) locking plate: e.g., a left distal femoral plate for femoral fracture on right-hip implant. This presents several advantages: minimally invasive surgery, the introduction of the plate being facilitated by the LISSTM ancillary; ease of locking, also thanks to the ancillary; and, above all, multiple proximal trochanteric fixation thanks to the form of this anatomic distal LISSTM plate, improving proximal bone fixation. The present technical note seeks to illustrate the interest of using a "reversed" plate, in terms of simplicity of fitting and quality of reduction and consolidation, while also specifying the associated limitations and tolerance. PMID- 21641899 TI - A metastatic case of gastrointestinal stromal tumor (GIST) treated with imatinib mesylate (Glivec((r))) in Mali. AB - Gastrointestinal stromal tumors (GIST) usually showing a spindle cells pattern of cell proliferation have recently benefit from a molecular definition. Indeed imatinib mesylate (Gleevec((r))) treatment has dramatically improved the management of these tumors as they frequently express the c-kit oncogene. We report the first case of a metastatic gastric GIST in a man of 45 years diagnosed and treated in Mali. The gastric tumor was particularly aggressive with a large intra-abdominal and mesenteric spreading and liver metastases. The diagnosis was done on the CD117 and CD34 expression in the tumor sample obtained by laparotomy. After a 34 months 400mg/day imatinib mesylate (Gleevec((r))) treatment a dramatic tumor regression was obtained. PMID- 21641900 TI - Evaluation of formalin-free tissue fixation for RNA and microRNA studies. AB - FineFix, RCL-2 and HOPE, three formalin-free fixatives, were compared to the common used formalin fixed tissue samples of lung cancer and were evaluated for their effects on quality, quantity and integrity of RNA and microRNA. Two commercially available RNA extraction Kits (RNeasy FFPE by Qiagen and RecoverAllTM Nucleic Acid Isolation by Ambion) were tested and optimized in order to determine an extraction protocol for RNA as well as miRNA independent of the fixative. Two selected miRNAs were quantified via TaqMan MicroRNA assays. The optimized RNA extraction protocol for Qiagen's Kit leads to similar results for RNA quality and integrity for all fixatives. Highest RNA yield was obtained for formalin and the highest average miRNA ratio was found for FineFix. RNA fragments smaller than 500 bases were detected in FineFix, formalin and RCL2 fixed tissues; HOPE was the only fixative showing long fragments in one third of the samples. Our findings demonstrate that formalin-free fixatives are in general not superior for RNA studies. With our optimized RNA extraction protocol, there is no difficulty in extracting great amounts of RNA with high quality. According to the quality obtained, quantitative real-time PCR analysis can be performed without any negative impact. Similar results can be achieved for the tested fixatives and therefore no fixative seems to represent a new "gold-standard" for tissue fixation. PMID- 21641901 TI - Claudin-7 inhibits human lung cancer cell migration and invasion through ERK/MAPK signaling pathway. AB - Tight junctions are the most apical component of the junctional complex critical for epithelial cell barrier and polarity functions. Although its disruption is well documented during cancer progression such as epithelial-mesenchymal transition, molecular mechanisms by which tight junction integral membrane protein claudins affect this process remain largely unknown. In this report, we found that claudin-7 was normally expressed in bronchial epithelial cells of human lungs but was either downregulated or disrupted in its distribution pattern in lung cancer. To investigate the function of claudin-7 in lung cancer cells, we transfected claudin-7 cDNA into NCI-H1299, a human lung carcinoma cell line that has no detectable claudin-7 expression. We found that claudin-7 expressing cells showed a reduced response to hepatocyte growth factor (HGF) treatment, were less motile, and formed fewer foot processes than the control cells did. In addition, cells transfected with claudin-7 dramatically decreased their invasive ability after HGF treatment. These effects were mediated through the MAPK signaling pathway since the phosphorylation level of ERK1/2 was significantly lower in claudin-7 transfected cells than in control cells. PD98059, a selective inhibitor of ERK/MAPK pathway, was able to block the motile effect. Claudin-7 formed stable complexes with claudin-1 and -3 and was able to recruit them to the cell-cell junction area in claudin-7 transfected cells. When control and claudin-7 transfected cells were inoculated into nude mice, claudin-7 expressing cells produced smaller tumors than the control cells. Taken together, our study demonstrates that claudin-7 inhibits cell migration and invasion through ERK/MAPK signaling pathway in response to growth factor stimulation in human lung cancer cells. PMID- 21641902 TI - Tribute to Rosario Hernandez. PMID- 21641903 TI - Focus on molecules: lutein. PMID- 21641904 TI - Host selection by the Louse Fly Crataerina pallida, an avian nest ectoparasite of the Common Swift Apus apus. AB - Preferences by parasites for particular hosts may have important implications for the functioning of host-parasite systems, however, this parasitic life-history trait remains little studied. No detrimental effect of Louse Fly Crataerina pallida parasitism has been found on Common Swift Apus apus nestling hosts. Host selection choices may be mediating the effect this parasite has and account for this apparent avirulence. Two aspects of parasite host selection were studied at a breeding colony of Common Swifts during 2008; (1) intra-brood differences in C. pallida parasitism were studied to determine the influence of nestling rank, (2) differences in male and female C. pallida parasitism were investigated, as they may result in varying costs of parasitism to hosts. C. pallida populations were found to preferentially parasitize higher rather than lower ranking nestlings within broods of both two and three chicks. Greater proportions of females were seen upon nestlings than at the nest, and upon higher ranking than lower ranking nestlings within broods. These results indicate that host selection occurs and this may thus account for the lack of parasitic virulence reported within this host-parasite system. PMID- 21641905 TI - Forty years of shortness of breath and lower extremity edema. Diagnosis: Primary intestinal lymphangiectasia (Waldmann's disease). PMID- 21641906 TI - An unusual cause of upper GI bleeding. Diagnosis: Pancreatic arteriovenous malformation. PMID- 21641907 TI - Comparative analysis of Met-enkephalin, galanin and GABA immunoreactivity in the developing trout preoptic-hypophyseal system. AB - We studied the organization of Met-enkephalin-containing cells and fibers in the developing preoptic-hypophyseal system of the brown trout (Salmo trutta fario) by immunohistochemistry and determined the relationship of these cells and fibers to the galaninergic and GABAergic systems. Met-enkephalin immunoreactivity was observed in cells in the preoptic area, the hypothalamus and the pituitary of late larvae. In the hypophysis, a few Met-enkephalin-containing cells were present in all divisions of the adenohypophysis, and some immunoreactive fibers were present in the interdigitations of the neural lobe with the proximal pars distalis. Concurrently, GABAergic fibers innervated the anterior and posterior neural lobe. Galanin cells coexisted with Met-enkephalin cells in neuronal groups of the preoptic-hypophyseal system. Galaninergic and GABAergic fibers innervated the preoptic and hypothalamic areas, but GABAergic fibers containing galanin were not observed. These results indicate that Met-enkephalin, galanin and GABA may modulate neuroendocrine activities in the preoptic area, hypothalamus and pituitary during the transition from larval to juvenile period. To better know how the development of the trout preoptic-hypophyseal system takes place, we studied the patterns of cell proliferation and expression of Pax6, a conserved transcription factor involved in the hypophysis development. Pax6 expressing cells and proliferating cells were present in the Rathke's pouch, the hypothalamus and the hypophysis of early larvae. In late larvae, Pax6 expression was no longer observed in these areas, and the density of proliferating cells largely decreased throughout development, although they remained in the hypophysis of late larvae and juveniles, suggesting that Pax6 might play an important role in the early regionalization of the pituitary in the trout. PMID- 21641908 TI - Efficacy of pharmacological estrogen receptor antagonists in blocking activation of zebrafish estrogen receptors. AB - A variety of pharmacological agonists, antagonists and selective estrogen receptor modulators (SERM) have been used to better understand the role of specific receptors in various physiological processes. Despite similar structure and function, less is known about the effect of agonists and antagonists on teleost estrogen receptors and the results of these studies have indicated wide variation among species. The goal of this study was to determine the ability of two human SERMs to modulate activation of three zebrafish estrogen receptor isoforms. Full length cDNA of zebrafish estrogen receptor 1 (esr1), estrogen receptor 2a (esr2a) and estrogen receptor 2b (esr2b) were cloned into expression vectors and transfected into cells that do not endogenously express any estrogen receptor along with an estrogen responsive luciferase vector. Cells transfected with any of the zebrafish estrogen receptors individually and then exposed to 17beta-estradiol (E2) or 17alpha-ethinylestradiol (EE2) exhibited a dose dependent increase in luciferase activity. None of the pharmacological antagonists, ICI 182, 780, methyl-piperidino-pyrazole (MPP) or pyrazolo [1,5-a] pyrimidine (PHTPP), were able to independently transactivate luciferase expression with any of the zebrafish estrogen receptors. Of the three ER antagonists, only ICI 182, 780 was able to block EE2 induced luciferase activity, although a 10 to 100-fold excess of ICI 182, 780 was necessary with all receptors. Neither MPP nor PHTPP were able to block EE2 induced luciferase activity with any isoform of zebrafish estrogen receptor. These results indicate that the difference between human ER and zebrafish ER ligand binding is not conserved enough for the SERMs MPP or PHTPP to elicit similar effects in zebrafish as those manifested in humans. PMID- 21641909 TI - Spatiotemporal patterns of IGF-like peptide expression in the silkmoth Bombyx mori predict its pleiotropic actions. AB - In vertebrates, insulin-like growth factors (IGFs) play important roles in the regulation of growth and development. Although the principal source of circulating IGFs is the liver, IGFs are also secreted by many other tissues, functioning locally through paracrine/autocrine mechanism. In the silkmoth Bombyx mori, Bommo-IGF-like peptide (BIGFLP) is the functional counterpart of vertebrate IGFs and is mainly produced by the fat body, a functional equivalent of the vertebrate liver and adipocytes. However, its production by other tissues has not yet been analyzed. In this study, we systematically surveyed the BIGFLP-producing tissues and stages by means of immunohistochemistry, in situ hybridization and real-time quantitative RT-PCR, showing that BIGFLP is also produced by the neurosecretory cells in the brain, ovariole sheath and testis sheath, in a stage specific manner. The BIGFLP-producing cells in the brain were identical to the cells that produce bombyxins, insulin-like peptides of B. mori, but the temporal expression patterns of both peptides were totally different. The BIGFLP gene expression in the sheaths of ovariole and testis were induced by ecdysteroid in vitro, similar to the expression in the fat body. A very high BIGFLP immunoreactivity was also found in the pupal nephrocytes, a functional equivalent of the glomerular podocytes in the vertebrate kidney, without the expression of the gene, suggesting that circulating BIGFLP is taken up and degraded by these tissues. Based on the present observations, the physiological functions of BIGFLP in B. mori development are discussed. PMID- 21641910 TI - Effects of developmental conditions on nestling American Kestrel (Falco sparverius) corticosterone concentrations. AB - How nestling birds respond to stressful situations may constitute an important survival component that has lasting developmental effects on the hypothalamic pituitary adrenal (HPA) axis. As birds are exposed to increasing amounts of potential anthropogenic stressors through land use change, understanding how these factors contribute to HPA development is important. We examined whether conditions experienced during the nestling stage affected free-living American Kestrel (Falco sparverius) HPA activity prior to fledging. Kestrels experienced varying levels of human disturbance around their nest and we classified this environmental exposure as high or low environmental human disturbance based on traffic patterns and land use. We then exposed some broods from high and low disturbance areas to a standardized disturbance protocol. Prior to fledging we collected blood samples from 25-day-old nestlings immediately after capture and 15 min post-capture. Corticosterone (CORT) did not vary with environmental human disturbance levels, disturbance protocol treatment, or with an interaction between environmental human disturbance and disturbance protocol treatment suggesting that nestling kestrels may not perceive external conditions related to human disturbance as stressful or kestrels may acclimate to disturbance. We also compared the relative effects of environmental human disturbance outside the nest cavity, conditions within the nest cavity (brood size), and individual condition (nestling fat scores) on baseline and stress-induced CORT. Baseline CORT did not vary with human disturbance level, brood size or fat score. Fat scores best explained stress-induced CORT with nestlings in better condition displaying elevated CORT. These results suggest that individual variation is more likely to explain HPA development compared to nest conditions or the external environment. This study demonstrates the importance of considering the effects of developmental conditions on the stress response at several scales. PMID- 21641911 TI - Detection of proliferative responses to ESAT-6 and CFP-10 by FASCIA assay for diagnosis of Mycobacterium tuberculosis infection. AB - There is a large and growing worldwide need for reliable tests to diagnose active and latent tuberculosis (TB). Improved methodology for identifying individuals with true latent TB (LTBI), particularly those with a recent infection, would pave the way for targeted prophylactic treatment. The traditionally used tuberculin skin test (TST) is unspecific and impractical. Interferon gamma release assays (IGRA) are more specific than the TST but, like that test, cannot discriminate either between recent and remote TB infection, or between these and a mere immunological memory of previous TB infection. The Flow-cytometric Assay for Specific Cell-mediated Immune-response in Activated whole blood (FASCIA) combines long-term antigen stimulation of whole blood and flow-cytometric analysis with quantification of the expanded T-lymphoblasts and can also be employed for measurement of cytokine responses. PMID- 21641912 TI - Lifetime gains and patterns of accumulation and mobilization of nutrients in females of the synovigenic parasitoid, Diglyphus isaea Walker (Hymenoptera: Eulophidae), as a function of diet. AB - In nature, adult parasitoids feed to obtain and use nutrients for supplementing and/or replenishing some of their existing array of nutrient reserves. When adults feed on host or non-host food, they can enhance fitness, typically by increasing egg production or longevity. In the present study, ovigeny index (OI) and impact of female fitness, as well as physiological state on the reproductive strategies, were investigated in the synovigenic parasitoid, Diglyphus isaea, fed on host food (2-3rd instars of Liriomyza sativae larvae), non-host foods (10% honey solution) and starved (distilled water, control). The results showed that D. isaea was a strongly synovigenic parasitoid, of which OI value was 0.002. Both types of food enhanced the fecundity and prolonged the longevity of the females. D. isaea females fed on non-host food showed higher levels of gut sugar, body sugar and glycogen than those fed on host food, but the levels of lipid were higher in the host-fed females. D. isaea females seemed to show lipogenesis, with low rates of lipid catabolism sufficient to satisfy the requirement of egg maturation. Females might absorb lipids directly from the haemolymph of paralyzed hosts. PMID- 21641913 TI - Three-dimensional structure of alpha-crystallin domain dimers of human small heat shock proteins HSPB1 and HSPB6. AB - Small heat shock proteins (sHSPs) are a family of evolutionary conserved ATP independent chaperones. These proteins share a common architecture defined by a signature alpha-crystallin domain (ACD) flanked by highly variable N- and C terminal extensions. The ACD, which has an immunoglobulin-like fold, plays an important role in sHSP assembly. This domain mediates dimer formation of individual protomers, which then may assemble into larger oligomers. In vertebrate sHSPs, the dimer interface is formed by the symmetrical antiparallel pairing of two beta-strands (beta7), generating an extended beta-sheet on one face of the ACD dimer. Recent structural studies of isolated ACDs from a number of vertebrate sHSPs suggest a variability in the register of the beta7/beta7 strand interface, which may, in part, give rise to the polydispersity often associated with the full-length proteins. To further analyze the structure of ACD dimers, we have employed a combination of X-ray crystallography and solution small-angle X-ray scattering (SAXS) to study the ACD-containing fragments of human HSPB1 (HSP27) and HSPB6 (HSP20). Unexpectedly, the obtained crystal structure of the HSPB1 fragment does not reveal the typical beta7/beta7 dimers but, rather, hexamers formed by an asymmetric contact between the beta4 and the beta7 strands from adjacent ACDs. Nevertheless, in solution, both ACDs form stable dimers via the symmetric antiparallel interaction of beta7 strands. Using SAXS, we show that it is possible to discriminate between different putative registers of the beta7/beta7 interface, with the results indicating that, under physiological conditions, there is only a single register of the strands for both proteins. PMID- 21641914 TI - Binding of two PriA-PriB complexes to the primosome assembly site initiates primosome formation. AB - A direct quantitative analysis of the initial steps in primosome assembly, involving PriA and PriB proteins and the minimal primosome assembly site (PAS) of phage phiX174, has been performed using fluorescence intensity, fluorescence anisotropy titration, and fluorescence resonance energy transfer techniques. We show that two PriA molecules bind to the PAS at both strong and weak binding sites on the DNA, respectively, without detectable cooperative interactions. Binding of the PriB dimer to the PriA-PAS complex dramatically increases PriA's affinity for the strong site, but only slightly affects its affinity for the weak site. Associations with the strong and weak sites are driven by apparent entropy changes, with binding to the strong site accompanied by a large unfavorable enthalpy change. The PriA-PriB complex, formed independently of the DNA, is able to directly recognize the PAS without the preceding the binding of PriA to the PAS. Thus, the high-affinity state of PriA for PAS is generated through PriA-PriB interactions. The effect of PriB is specific for PriA-PAS association, but not for PriA-double-stranded DNA or PriA-single-stranded DNA interactions. Only complexes containing two PriA molecules can generate a profound change in the PAS structure in the presence of ATP. The obtained results provide a quantitative framework for the elucidation of further steps in primosome assembly and for quantitative analyses of other molecular machines of cellular metabolism. PMID- 21641915 TI - Strategy abundance in evolutionary many-player games with multiple strategies. AB - Evolutionary game theory is an abstract and simple, but very powerful way to model evolutionary dynamics. Even complex biological phenomena can sometimes be abstracted to simple two-player games. But often, the interaction between several parties determines evolutionary success. Rather than pair-wise interactions, in this case we must take into account the interactions between many players, which are inherently more complicated than the usual two-player games, but can still yield simple results. In this manuscript we derive the composition of a many player multiple strategy system in the mutation-selection equilibrium. This results in a simple expression which can be obtained by recursions using coalescence theory. This approach can be modified to suit a variety of contexts, e.g. to find the equilibrium frequencies of a finite number of alleles in a polymorphism or that of different strategies in a social dilemma in a cultural context. PMID- 21641916 TI - Structural sensitivity of biological models revisited. AB - Enhancing the predictive power of models in biology is a challenging issue. Among the major difficulties impeding model development and implementation are the sensitivity of outcomes to variations in model parameters, the problem of choosing of particular expressions for the parametrization of functional relations, and difficulties in validating models using laboratory data and/or field observations. In this paper, we revisit the phenomenon which is referred to as structural sensitivity of a model. Structural sensitivity arises as a result of the interplay between sensitivity of model outcomes to variations in parameters and sensitivity to the choice of model functions, and this can be somewhat of a bottleneck in improving the models predictive power. We provide a rigorous definition of structural sensitivity and we show how we can quantify the degree of sensitivity of a model based on the Hausdorff distance concept. We propose a simple semi-analytical test of structural sensitivity in an ODE modeling framework. Furthermore, we emphasize the importance of directly linking the variability of field/experimental data and model predictions, and we demonstrate a way of assessing the robustness of modeling predictions with respect to data sampling variability. As an insightful illustrative example, we test our sensitivity analysis methods on a chemostat predator-prey model, where we use laboratory data on the feeding of protozoa to parameterize the predator functional response. PMID- 21641917 TI - Glioblastoma cells: a heterogeneous and fatal tumor interacting with the parenchyma. AB - Glioblastomas (GBMs) are considered to be one of the deadliest human cancers, characterized by a high proliferative rate, aggressive invasiveness and insensitivity to radio- and chemotherapy, as well as a short patient survival period. Moreover, GBMs are among the most vascularized and invasive cancers in humans. Angiogenesis in GBMs is correlated with the grade of malignancy and is inversely correlated with patient survival. One of the first steps in tumor invasions is migration. GBM cells have the ability to infiltrate and disrupt physical barriers such as basement membranes, extracellular matrix and cell junctions. The invasion process includes the overexpression of several members of a super-family of zinc-based proteinases, the Metzincin, in particular a sub group, metalloproteinases. Another interesting aspect is that, inside the GBM tissue, there are up to 30% of microglia or macrophages. However, little is known about the immune performance and interactions of the microglia with GBMs. These singular properties of GBMs will be described here. A sub-population of cells with stem-like properties may be the source of tumors since, apparently, GBM stem cells (GSCs) are highly resistant to current cancer treatments. These cancer therapies, while killing the majority of tumor cells, ultimately fail in GBM treatment because they do not eliminate GSCs, which survive to regenerate new tumors. Finally, GBM patient prognostic has shown little improvement in decades. In this context, we will discuss how the membrane-acting toxins called cytolysins can be a potential new tool for GBM treatment. PMID- 21641918 TI - Differential nuclear localisation and promoter occupancy play a role in glucocorticoid receptor ligand-specific transcriptional responses. AB - The glucocorticoid receptor (GR) is a ligand-activated transcription factor for which a number of endogenous and synthetic ligands exist. A key question in steroid receptor biology is how different ligands elicit different maximal transcriptional responses via the same receptor and on the same promoter. This question was addressed quantitatively for the GR, using a panel of agonists, partial agonists and antagonists, on the endogenous GILZ gene in two different human cell lines. It was found that the extent of GR nuclear localization correlated with the efficacy for GILZ transactivation by the GR in U2OS cells. However, in A549 cells there was no significant correlation, with all ligands resulting in similar levels of GR nuclear localization, despite different levels of transcriptional activation of the GILZ gene. Chromatin immunoprecipitation analysis on the other hand, revealed ligand-specific differences in GILZ promoter occupancy in the A549 cells, which correlated with the transcriptional efficacy of the subset of ligands investigated. This suggests that ligand-specific differences in promoter occupancy by activated GR play a major role in discrimination between agonist, partial agonist and antagonist responses on the endogenous GILZ gene in A549 cells, while differences in nuclear localisation of liganded GR play a role in determining the transcriptional outcome in U2OS cells. These cell line-specific differences were not dependent on the amount of GR present, since transient overexpression of GR in U2OS did not alter the relative ligand-selective nuclear localisation. Our results show that there is a relationship between ligand-specific transactivation efficacy, extent of nuclear translocation and recruitment of GR to the promoter. However, the relative contribution of nuclear translocation and GR promoter recruitment to ligand specific transactivation efficacy is cell-specific. PMID- 21641919 TI - Cytotoxic steroids from Monascus purpureus-fermented rice. AB - Bioassay-guided fractionation of an EtOH extract of Monascus purpureus-fermented rice led to the isolation of two new steroids (22S, 23R, 24S) 20beta,23alpha,25alpha-trihydroxy-16,22-epoxy-4,6,8(14)-trienergosta-3-one (1), the first example of a steroid possessing both a conjugated triene ketone system and a fused 4H-furan ring side chain within one molecule, and (22E, 24R) 3beta,5alpha-dihydroxyergosta-23-methyl-7,22-dien-6-one (2), as well as two known compounds (22E, 24R)-3beta,5alpha-dihydroxyergosta-7,22-dien-6-one (3) and (22E, 24R)-6beta-methoxy-ergosta-7,22-diene-3beta,5alpha-diol (4). Their structures were assigned by detailed interpretation of HRESIMS, 1D and 2D NMR spectroscopic data. The absolute stereochemistry of 1 was determined by single-crystal X-ray crystallography while the absolute stereochemistry of 2 was established by CD. Compounds 1-4 showed cytotoxic activity against the lung adenocarcinoma (A549) with IC(50) values of 0.08, 0.94, 12.6 and 13.5 MUM, respectively. In addition, compounds 1 and 2 exhibited moderate activities against human ovarian cancer (A2780), with IC(50) values of 2.8 and 5.1 MUM. PMID- 21641920 TI - Chronopotentiometric studies of phosphatidylcholine bilayers modified by ergosterol. AB - We have monitored the effect of ergosterol on electrical capacitance and electrical resistance of the phosphatidylcholine bilayer membranes using chronopotentiometry method. The chronopotentiometric characteristic of the bilayers depends on constant-current flow through the membranes. For low current values, no electroporation takes place and the membrane voltage rises exponentially to a constant value described by the Ohm's law. Based on these kinds of chronopotentiometric curves, a method of the membrane capacitance and the membrane resistance calculations is presented. PMID- 21641921 TI - Leucurogin, a new recombinant disintegrin cloned from Bothrops leucurus (white tailed-jararaca) with potent activity upon platelet aggregation and tumor growth. AB - Disintegrins and disintegrins-like proteins are able to inhibit platelet aggregation and integrin-mediated cell adhesion. The aim of this study was to produce one disintegrin-like cloned from Bothrops leucurus venom gland and to characterize it regarding biological activity. The recombinant protein was purified by one step procedure involving anion-exchange chromatography (DEAE cellulose) and presented a molecular mass of 10.4 kDa. The purified protein was able to inhibit platelet aggregation induced by collagen (IC50 = 0.65 MUM) and to inhibit growth of Ehrlich tumor implanted in mice by more than 50% after 7 days administration of 10 MUg/day. No effects were observed upon adenosine 5' diphosphate (ADP)-and arachidonic acid (AA)-induced platelet aggregation. The recombinant protein was recognized by an antibody specific for jararhagin one metalloproteinase isolated from Bothrops jararaca venom, and therefore it was named leucurogin. Anti-angiogenesis effect of leucurogin was evaluated by the sponge implant model. After 7 days administration leucurogin inhibited, in a dose dependent way, the vascularization process in the sponge. Leucurogin represents a new biotechnological tool to understand biological processes where disintegrins like are involved and may help to characterize integrins that can be involved in development and progression of malignant cells. PMID- 21641922 TI - Pharmacokinetic and pharmacodynamic interactions of valproate, phenytoin, phenobarbitone and carbamazepine with curcumin in experimental models of epilepsy in rats. AB - The present study investigates the interaction of curcumin with four antiepileptic drugs (AEDs) in male Wistar rats. In the first protocol, seizures were induced using pentylenetetrazole (PTZ) and valproate was injected intraperitoneally (i.p.) in therapeutic and sub-therapeutic doses 30min before PTZ administration. Curcumin was co-administered with sub-therapeutic dose of valproate 60min before PTZ injection. In the second protocol, seizures were induced by maximal-electroshock. Phenytoin, phenobarbitone and carbamazepine were injected in their therapeutic and sub-therapeutic doses 120, 60 and 30min, respectively, before seizure induction. Curcumin was administered along with sub therapeutic doses of phenytoin, phenobarbitone and carbamazepine, 60min before induction of seizures. Behavioral parameters were assessed using elevated plus maze test and passive avoidance paradigm. Rat brain oxidative stress parameters were assessed and the serum levels of the AEDs were estimated. The AEDs in their therapeutic doses produced complete protection against seizures. However, sub therapeutic doses of these AEDs failed to completely protect against seizures. Co administration of curcumin with sub-therapeutic dose of valproate significantly increased the latency to myoclonic jerks. The percentage protection against seizures with sub-therapeutic doses of valproate, phenytoin, phenobarbitone and carbamazepine was also enhanced by concomitant curcumin administration. Both PTZ and MES induced seizures caused significant impairment of cognitive functions. Co administration of curcumin with these AEDs in their sub-therapeutic doses prevented the impairment of learning and memory due to seizures whereas no such improvement was observed in the groups administered the sub-therapeutic doses of the AEDs alone. Additionally, curcumin reversed the oxidative stress due to seizures. However, curcumin co-administration did not cause any significant alteration in the serum levels of the AEDs. The results thus suggest the potential of curcumin as an adjunct to these AEDs in epilepsy with the advantage of increasing the efficacy, reducing the dose and side effects of the AEDs. PMID- 21641923 TI - Mineralization of pristine chitosan film through biomimetic process. AB - The biomineralization of pristine chitosan film without any prior surface treatment was evaluated by immersing the film in simulated body fluid (SBF) at 37 degrees C for 3 weeks. The film was prepared by solvent casting method using chitosan of known degree of deacetylation (DD). The formation of the hydroxyapatite (HA) phase on the film surface after immersion was studied periodically by X-ray powder diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), energy dispersive X-ray analysis (EDX) and scanning electron microscopy (SEM) methods. The electron micrographs showed the morphology of the deposited apatite as small globules appearing uniformly throughout the films surfaces. The Ca/P ratio of the apatite was found to increase with increase in immersion time and approaching towards the stoichiometric value of the HA phase. The mineralized chitosan film could be of promising support to hard tissue regeneration. PMID- 21641924 TI - Controlled release of tamoxifen citrate encapsulated in cross-linked guar gum nanoparticles. AB - Natural polysaccharides, due to their outstanding merits, have received more and more attention in the field of drug delivery. In the present study tamoxifen citrate, TMX (a non-steroidal antiestrogenic drug) loaded guar gum nanoparticles, GG NPs, crosslinked with glutaraldehyde were prepared for treatment of breast cancer. An oil in water (o/w) emulsion polymer cross-linking method was employed for preparation of blank and drug loaded sustained release nature biodegradable nanoparticles. Prepared nanoparticles were characterized by morphology in scanning electron microscope (SEM), size distribution in transmission electron microscope (TEM), TMX loading by high performance liquid chromatography (HPLC) and in vitro drug release characteristics. An overall sustained release of the drug from the biodegradable nanoparticles was observed in in vitro release studies. The release of TMX from GG NPs was found to be effected by guar gum and glutaraldehyde concentration. Regression coefficient (R(2)) analysis suggested that the predominant mechanism behind the drug release from the nanoparticles was time dependent release and diffusion. In vivo studies on female albino mice demonstrated maximum uptake of the drug by mammary tissue after 24h of administration with drug loaded guar gum nanoparticles in comparison with that with the tablet form of the drug. These findings demonstrate that controlled release of TMX from GG NPs could be a potential alternative pharmaceutical formulation in passive targeting of TMX in breast cancer treatments. PMID- 21641925 TI - Glycosyl part identified within Balanites aegyptiaca fruit protease. AB - The many milk-clotting proteases from plant are glycosylated; attachment of monosaccharides to enzyme is an advantage for its activity and stability. In this study, gas chromatography coupled to mass spectrometry-electrospray ionization was used to identify glycans bond to proteases purified from Balanites aegyptiaca fruits pulp through cation exchange chromatography. Carbohydrates were identified according to the retention time and the ion at m/z after derivation by heptafluorobutyric acid. The chromatograms obtained from monosaccharides analysis revealed the presence of galactose, mannose, arabinose, xylose, rhamnose and glucuronic acid. The mass spectrometry-electrospray ionization spectra corroborated these findings. PMID- 21641926 TI - Loss of Hemolectin reduces the survival of Drosophila larvae after wounding. AB - Coagulation involving both hemocytes and humoral factors is important for insect survival and immune defense. Hemolectin is a major larval clotting factor in Drosophila, and hemolymph from hml mutants does not clot ex vivo. Yet surprisingly third instar hml larvae survived injury as well as controls. The number of hemocytes in circulation changes during larval development. Reasoning that this could affect coagulation, we studied larval survival after injury at different stages. We found that hml larvae survived less than controls when injured during the feeding stage with fewer hemocytes. This important in vivo result reinforces the role of Hemolectin in larval hemostasis. A subtle effect of hml on immunity was found in adults. Similar experiments on hml mutant larvae gave different results, but feeding stage hml larvae were differentially sensitive to infections with different strains of Serratia marcescens. PMID- 21641927 TI - Adrenergic regulation of the innate immune response in common carp (Cyprinus carpio L.). AB - Catecholamines exert their physiological actions through alpha and beta adrenergic receptors (ARs). As ARs are not exclusively expressed on neuroendocrine cells, but also on leukocytes, they may facilitate neuroendocrine modulation of immune responses. We sequenced the beta(2a)-AR in common carp, and studied its expression profile and involvement in the regulation of teleost innate immune responses. beta(2a)-AR messenger RNA was found to be constitutively expressed in brain areas, especially in the preoptic nucleus (NPO, homologous to the mammalian hypothalamus), and in immune organs. During the active phase of an in vivo inflammatory response, induced by i.p. zymosan treatment, beta(2a)-AR gene expression was up-regulated in the peritoneal leukocytes. Additionally, adrenaline in vitro reduced the synthesis of oxygen radical species and nitric oxide, while it enhanced arginase activity in fish phagocytes. Furthermore, in vitro adrenaline administration inhibited expression of pro-inflammatory cytokines, chemokines and their receptors. It is therefore hypothesized that adrenaline will down-regulate phagocyte skewing toward classical/innate polarization. PMID- 21641928 TI - A holistic view of the dynamisms of teleost IgM: a case study of Streptococcus iniae vaccinated rainbow trout (Oncorhynchus mykiss). AB - To date, little is known about how trout IgM, the primary antibody of fish, varies in titer, specificity, disulfide cross-linking, and affinity following immunization with a pathogen. Work using defined antigens has demonstrated that the disulfide cross-linking structure of IgM becomes increasingly more polymerized during an immune response, coinciding with an increase in affinity, but it is unknown if this has relevance to aquatic pathogens. Understanding how IgM varies following vaccination with an aquatic pathogen is of considerable importance as effector functions allocated to multiple antibody isotypes in mammals are essentially relegated to this single molecule. To gain insights into the dynamism of IgM, rainbow trout were immunized with Streptococcus iniae and individual serum titers, their specificity and affinity to S. iniae, and the disulfide cross-linking pattern of both total-serum and specific Ig were analyzed over a period of 37 weeks. We found that in vaccinated animals titer increased by a factor of ~100 from starting levels, affinity increased 10-fold, and diversity of S. iniae proteins recognized by trout antibody increased at least 5-fold. Most intriguing, though less cross-linked IgM predominated early in response, by week 5, the fully tetramerized antibody comprised 50% of total specific protein. We propose that this is a mechanism to optimize efficacy of carrying out effector functions and recognizing a wide array of epitopes with higher affinity. PMID- 21641929 TI - Identification of putative miRNA involved in Drosophila melanogaster immune response. AB - MicroRNAs (miRNAs) are small endogenous non-coding RNAs that post transcriptionally regulate gene expression in eukaryotes. They are known to play diverse roles in physiological processes such as homeostasis, development, cancer and immune response. In Drosophila melanogaster up to 176 miRNAs have been identified; yet, their biological functions remain unknown. Here, we describe an in silico screening strategy to identify miRNAs involved in a specific immune signaling pathway that is based on: (i) the potential capability of miRNAs to target mRNAs of a given pathway; (ii) the sequence conservation of miRNAs across species and (iii) the expression profile of miRNAs. Using this strategy, we have defined a subset of seven Drosophila miRNAs that are likely to participate in the immune response. Interestingly, some of these miRNAs target peptidoglycan receptor proteins (PGRPs) for which no regulators are known yet. miRNA-mediated regulation may explain how PGRPs are controlled in the immune signaling pathway. PMID- 21641930 TI - The inhibitory effect of environmental ammonia on Danio rerio LPS induced acute phase response. AB - Ammonia is a toxic by-product of amino acid catabolism and a common environmental pollutant that has been associated with increased disease susceptibility in fish although the mechanism is not well understood. We addressed the hypothesis that elevated environmental ammonia acts by impairing the acute phase response (APR). Specifically, we determined the impact of sub-lethal acute (24 h) and chronic (14 d) ammonia exposure on acute phase protein gene expression in zebrafish (Danio rerio) in response to a challenge with bacterial lipopolysaccharide (LPS: i.p. 10 MUg/g after 24h). A panel of LPS-responsive genes (SAA, HAMP, LECT2, Hp and IL1beta) were identified and evaluated by real-time quantitative PCR. Ammonia was found to impair induction of SAA, HAMP and LECT2 by 50-90%. Both short (15 min, 1h and 24h) and long-term (14 days) exposure to high environmental ammonia concentrations significantly elevated whole-body cortisol levels compared with control fish. Our results reveal for the first time that exposure to high environmental levels of ammonia suppresses the innate immune response in fish. We hypothesize that high environmental ammonia-mediated elevation of cortisol levels in zebrafish may be playing a key role in this immunosuppression, while the mechanisms involved remains to be elucidated. PMID- 21641931 TI - Conjunctiva-associated lymphoid tissue in avian mucosal immunity. AB - Conjunctiva-associated lymphoid tissue's (CALT) role in generating avian mucosal adaptive immunity was measured by analyzing cellular composition, expression of the polymeric immunoglobulin receptor (pIgR), and production of cytokines and antibodies in chickens ocular exposed to a replication-deficient adenovirus of serotype 5 (Ad5). These studies demonstrate that CALT contains B cells, gammadelta T cells, T helper, and cytotoxic T cells, and a T lymphocyte composition, which more resembles Harderian glands than spleen. CALT-derived lymphocytes contain antigen-specific, IgA-secreting plasma cells and cytokine producing lymphocytes after ocular Ad5 vaccination. The expression of the pIgR in the CALT's lymphoepithelium emphasizes the importance of mucosal immune protection by paraocular lymphoid tissues. The CALT immune response after ocular Ad5 boosting was influenced by prior high dose in ovo Ad5 priming. Thus, both mucosal and systemic immunization influenced Ad5-induced IFN-gamma responses in CALT. PMID- 21641932 TI - Gender and manganese exposure interactions on mouse striatal neuron morphology. AB - Gender differences in sensitivity and toxicokinetics of multiple metals have been identified in humans. A recent study suggested that young girls performed worse on intellectual exams than young boys exposed to manganese (Mn) in the environment. Animal studies have shown that Mn exposure causes differential effects on behavior in male compared to female mice. We hypothesized that in response to Mn exposure striatal Mn accumulation and/or striatal medium spiny neuron (MSN) morphology show gender-dependent effects. We evaluated the contribution of gender to neuropathology by examining striatal MSN morphology in male and female mice exposed to Mn. We found that gender played a significant role in alterations of striatal MSN morphology in mice exposed to Mn. Gender dependent changes were strongest when striatal Mn levels were elevated 24h following the final Mn exposure. Nevertheless, gender-dependent alterations in neuron morphology were still present 3 weeks after the final Mn exposure. Gender differences in neuron morphology were not due to differential striatal Mn accumulation between genders. We conclude that although gender does not affect striatal Mn accumulation, MSN morphology is differentially sensitive to elevated Mn levels. PMID- 21641933 TI - Diet composition modifies the toxicity of repeated soman exposure in rats. AB - It was previously demonstrated that diet potently modulates the toxic effects of an acute lethal dose of the nerve agent soman. The current investigation was undertaken to examine the influence of diet on the cumulative toxicity of repeated soman administration. Rats were fed one of four distinct diets (standard, choline-enriched, glucose-enriched, or ketogenic) for four weeks prior to and throughout a repeated soman dosing and recovery regimen. Each diet group included animals exposed to an equivalent volume of saline that served as negative controls. In exposure Week 1, animals received three consecutive daily doses of 0.4 LD(50) soman. In exposure Week 2, animals received four consecutive daily doses of 0.5 LD(50) soman. In exposure Week 3, animals received five consecutive daily doses of 0.5 LD(50) soman. Week 4 constituted a post-exposure recovery evaluation. Throughout the experiment, behavioral function was assessed by a discriminated avoidance test that required intact sensory and motor function. Survival and body weight changes were recorded daily. Differences in toxicity as a function of diet composition became apparent during the first week. Specifically, rats fed the glucose-enriched diet showed pronounced intoxication during Week 1, resulting in imperfect survival, weight loss, and deteriorated avoidance performance relative to all other groups. All rats fed the glucose enriched diet died by the end of exposure Week 2. In contrast, only 10% of animals fed the standard diet died by the end of Week 2. Also in Week 2, weight loss and disrupted avoidance performance were apparent for all groups except for those fed the ketogenic diet. This differential effect of diet composition became even more striking in Week 3 when survival in the standard and choline diet groups approximated 50%, whereas survival equaled 90% in the ketogenic diet group. Avoidance performance and weight loss measures corroborated the differential toxicity observed across diet groups. Upon cessation of soman exposure during the final week, recovery of weight and avoidance performance in survivors was comparable across diet groups. These results systematically replicate previous findings demonstrating that diet composition exacerbates or attenuates toxicity in rodents exposed acutely to organophosphorus compounds. PMID- 21641934 TI - Sparse geostatistical analysis in clustering fMRI time series. AB - Clustering is used in fMRI time series data analysis to find the active regions in the brain related to a stimulus. However, clustering algorithms usually do not work well for ill-balanced data, i.e., when only a small proportion of the voxels in the brain respond to the stimulus. This is the typical situation in fMRI--most voxels do not, in fact, respond to the specific task. We propose a new method of sparse geostatistical analysis in clustering, which first uses sparse principal component analysis (SPCA) to perform data reduction, followed by geostatistical clustering. The proposed method is model-free and data-driven; in particular it does not require prior knowledge of the hemodynamic response function, nor of the experimental paradigm. Our data analysis shows that the spatial and temporal structures of the task-related activation produced by our new approach are more stable compared with other methods (e.g., GLM analysis with geostatistical clustering). Sparse geostatistical analysis appears to be a promising tool for exploratory clustering of fMRI time series. PMID- 21641935 TI - Immunological function of the endothelial cell within the setting of organ transplantation. AB - In organ transplantation, development of immunosuppressive treatment and improved diagnosis of allograft rejection has resulted in increased allograft survival in recent years. Nevertheless, rejection remains a major cause of graft loss and a better understanding of the characteristics of the allo-immune response is required to identify new diagnostic and therapeutic tools. The allogeneic immune response depends upon a major family of antigenic targets: the Major Histocompatibility Complex molecules (MHC) which are present on donor cells. These molecules are targets of both the humoral and cellular arms of the graft recipient's immune system: T lymphocytes which are implicated in acute cellular rejection and antibodies which are implicated in antibody-mediated rejection (AMR). Allo-recognition of allograft MHC antigens by either T cells or allo antibodies is the primary event which can ultimately lead to graft rejection. Although immunosuppressive strategies have mainly focused on the T cell response and acute cellular rejection has therefore become relatively rare, antibody mediated rejection (AMR) remains resistant to conventional immunosuppressive treatment and results in frequent graft loss. Damage to the endothelium is a prominent histological feature of AMR underlining the involvement of endothelial cells in initiating the allo-immune response. Furthermore, endothelial cells express both HLA class I and class II molecules in the context of organ transplantation endowing them with the capacity to present antigen to the recipient T cells. The endothelium should therefore be viewed both as a stimulator of, and as a target for allo-immune responses. In this review, we will summarize current knowledge about the implication of endothelial cells in the allo-immune response in the context of organ transplantation. PMID- 21641936 TI - Inhibition of influenza virus-induced NF-kappaB and Raf/MEK/ERK activation can reduce both virus titers and cytokine expression simultaneously in vitro and in vivo. AB - Influenza virus (IV) infection can cause severe pneumonia and death. Therapeutic actions are limited to vaccines and a few anti-viral drugs. These target viral functions thereby selecting resistant variants. During replication IV activates the Raf/MEK/ERK-cascade and the transcription factor NF-kappaB. Both result in virus supportive and anti-viral effects by promoting viral genome transport for virus assembly and by inducing expression of pro-inflammatory host factors. Apart from tissue damage caused by the virus lytic replication, an imbalanced overproduction of anti-viral cytokines can cause severe lung damage as observed in human H5-type IV infections. Recently we showed that inhibition of NF-kappaB activity reduces the virus titer in vitro and in vivo. We have now analyzed whether inhibition of these pathways, allows simultaneous reduction of virus titers and virus-induced cytokines. The results show that inhibition of either pathway indeed leads to decreased virus titers and cytokine expression. This was not only true for infected permanent cells or primary mouse alveolar epithelial cells, but also in infected mice. Hereby we demonstrate for the first time in vitro and in vivo that virus titers and pro-inflammatory cytokine expression can be modulated simultaneously. This could provide a new rationale of future therapeutic strategies to treat IV pneumonia. PMID- 21641937 TI - Dissociations within short-term memory in GluA1 AMPA receptor subunit knockout mice. AB - GluA1 AMPA receptor subunit knockout mice display a selective impairment on short term recognition memory tasks. In this study we tested whether GluA1 is important for short-term memory that is necessary for bridging the discontiguity between cues in trace conditioning. GluA1 knockout mice were not impaired at using short term memory traces of T-maze floor inserts, made of different materials, to bridge the temporal gap between conditioned stimuli and reinforcement during appetitive discrimination tasks. Thus, different aspects of short-term memory are differentially sensitive to GluA1 deletion. This dissociation may reflect processing of qualitatively different short-term memory traces. Memory that results in performance of short-term recognition (e.g. for objects or places) may be different from the memory required for associative learning in trace conditioning. PMID- 21641938 TI - Chlordiazepoxide and lavender oil alter unconditioned anxiety-induced c-fos expression in the rat brain. AB - Lavender oil has a long history of use for treating anxiety, but only recent research has examined its effects using standard behavioural methods used to test novel drugs. The aim of this study was to investigate the effects of inhaled lavender oil on anxiety related behaviour of rats in the open field and to compare them with the effects of chlordiazepoxide (CDP), a typical anxiolytic drug. Additionally c-fos immunochemistry was used to investigate whether lavender oil produced the same pattern of c-fos expression as CDP in eight different brain areas associated with anxiety. As previously found, lavender oil showed anxiolytic properties in the open field similar to but not as extensive as those of CDP. Immunochemistry results indicated that exposure to the open field increased c-fos expression, while CDP reversed the effects of this behavioural stressor on c-fos expression in all brain regions examined except the central nucleus of the amygdala, where c-fos expression increased. Lavender oil had similar effects to CDP on the paraventricular nucleus of the hypothalamus, the dorsomedial hypothalamic nucleus and the central nucleus of the amygdala. These results strengthen the suggestion that inhaling lavender oil has anxiolytic behavioural effects, but they are weaker than the effects of benzodiazepines, and there is limited evidence that they are mediated by the same neural processes. PMID- 21641939 TI - Serodiagnosis of environmental mycobacterial infections. AB - To demonstrate the usefulness of enzyme-linked immunosorbent assay for serodiagnosis of mycobacterioses due to environmental mycobacteria we utilized a panel of glycolipid antigens selective for Mycobacterium avium-intracellulare, Mycobacterium kansasii, Mycobacterium xenopi, Mycobacterium scrofulaceum and Mycobacterium gordonae. The levels of circulating antibodies were determined against the environmental mycobacteria, and Mycobacterium tuberculosis in human immunodeficiency virus-negative and -positive patient sera. The method used immunomagnetic separation of the antigens, with covalent immobilization of antibodies to superparamagnetic amine and carboxyl terminated particles in solutions of the specific antigens. Enzyme-linked immunosorbent assay was performed on 195 patient sera: 34 with infections due to environmental mycobacteria, 114 with tuberculosis, 47 with other respiratory diseases. There were 46 human immunodeficiency virus-1 infected individuals. Among the 34 infections due to environmental mycobacteria, 9 patients were singularly infected with an environmental mycobacterium, and 25 co-infected with both M. tuberculosis and an environmental mycobacterium. Sensitivity, specificity and false positivity ranges were determined for each of the volunteer groups: tuberculosis positive, human immunodeficiency virus negative; tuberculosis positive, human immunodeficiency virus positive; those with infections due to individual environmental mycobacteria (such as M. scrofulaceum and M. kansasii); and those with other respiratory diseases. We demonstrate that such multiple assays, can be useful for the early diagnosis of diverse environmental mycobacterial infections to allow the start of treatment earlier than henceforth. PMID- 21641940 TI - Developments in immunological technologies leading to improvements in point-of care diagnostic testing. AB - Rapid and accurate point of care testing is of great concern, especially in terms of detecting infectious disease outbreaks in developing countries having high population burdens. While numerous detection systems are currently available, care must be taken in choosing those that are reliable and have proven acceptable levels of sensitivity and specificity, based on adequate laboratory and pre clinical trial testing. Two papers in the current issue show significant advances in technology that could bring Point-of-Care Diagnostic Testing closer to reality for cholera and environmental mycobacteria. PMID- 21641941 TI - Response inhibition of children with ADHD in the stop-signal task: an event related potential study. AB - The Stop-Signal Task (SST) is a procedure that can provide a measure of inhibitory control of an ongoing motor response. We used the stop-signal paradigm to determine whether deficient inhibitory control distinguishes children with a diagnosis of attention deficit hyperactivity disorder combined type (ADHD-Com) from normally developing children, matched on age and sex. Participants performed a standard visual two-choice task with an auditory stop-signal stimulus, while an EEG was recorded. The behavioral results indicated that the ADHD group had significantly impaired inhibitory control compared to the performance of normal children. Relative to controls, the go stimulus reaction time (RT) and the stop signal reaction time (SSRT) were prolonged in children with ADHD. The amplitudes of P2 and P3 components to the auditory stop-signal were more pronounced for successful than for unsuccessful stop-signal trials in both groups. However, the successful-unsuccessful difference was larger in control subjects. In contrast, the amplitude of the N2 component to auditory stop-signal was more pronounced for unsuccessful than for successful stop-signal trials in both groups. The comparison of the N2 component between control and ADHD groups revealed a greater amplitude and longer latency in the latter group, in successful stop-signal trials only. Additionally, the amplitude of response-locked ERPs, containing the ERN-Pe complex related to error-detection, was smaller in ADHD children. These results support the hypothesis of a complex deficit of inhibitory control, conflict monitoring, and error recognition mechanisms in ADHD and corroborate the utility of the stop-signal task in distinguishing hyperactive from normal children. PMID- 21641942 TI - Attentional processes in children with ADHD: an event-related potential study using the attention network test. AB - A variety of event-related potential (ERP) based studies have shown differences in neuronal processes underlying attention, inhibition and error processing in children with attention-deficit/hyperactivity disorder (ADHD) compared to controls. However, so far there are no studies that have compared children with ADHD and typically developing (TD) children regarding effects in ERP components associated with the attention network test (ANT). The ANT allows to differentiate between three particular aspects of attention: alerting, orienting, conflict. Twenty-five children with ADHD and 19 TD children (comparable with respect to age, sex, and IQ) performed the ANT while ERPs were recorded. Based on DSM-IV, the group of children with ADHD was divided in an inattentive (ADHDin, n=10) and a combined (ADHDcom, n=15) subgroup. On the performance level, the ADHD group showed a significantly higher variability of reaction times. Concerning ERP measures, smaller cue-P3 amplitudes were found in the ADHD group indicating that children with ADHD allocate less attentional resources for cue processing. In addition, the target-P3 in ADHD showed smaller amplitudes. Subgroup analysis revealed reduced cue-P3 amplitudes in both subgroups and reduced target-P3 amplitudes in ADHDin compared to TD children. Except for a higher alerting score in ADHD after correction for cue-P3 group differences, performance data revealed no group differences specific for the three attention networks. No group differences related to the attention networks were observed at the ERP level. Our results suggest that deviant attentional processing in children with ADHD is only partly related to ANT-specific effects. Findings are compatible with the model of a suboptimal energetic state regulation in ADHD. Furthermore, our results suggest that deviant cue processing in ADHD and related differences in task modulations should be accounted for in data analysis. PMID- 21641943 TI - LORETA analysis of three-dimensional distribution of delta band activity in schizophrenia: relation to negative symptoms. AB - We sought to determine if altered electroencephalography (EEG) activities, such as delta band activity, in specific brain regions are associated with psychotic symptoms. Data were obtained from 17 neuroleptic-naive patients with schizophrenia and age- and sex-matched 17 healthy control subjects. Low Resolution Brain Electromagnetic Tomography (LORETA) was used to generate current source density images of delta, theta, alpha, and beta activities. Localization of the difference in EEG activity between the two groups was assessed by voxel-by voxel non-paired t-test of the LORETA images. Spearman's correlation coefficient was obtained to relate LORETA values of EEG current density in brain regions showing a significant between-group difference and psychopathology scores. Delta band activity, represented by LORETA current density, was greater for patients in the following areas; the left inferior temporal gyrus, right middle frontal gyrus, right superior frontal gyrus, right inferior frontal gyrus, and right parahippocampal gyrus. LORETA values for delta band activity in the above five brain regions were negatively correlated with negative, but not positive symptoms. The results of this study suggest the role for electrophysiological changes in some of the brain regions, e.g. prefrontal cortex, in the manifestation of negative symptoms. PMID- 21641944 TI - Construction and biological characterisation of recombinant porcine circovirus type 2 expressing the V5 epitope tag. AB - Porcine circovirus type 2 (PCV2) is a major causal agent of post-weaning multisystemic wasting syndrome in piglets. To investigate the feasibility of PCV2 expressing an exogenous epitope, a 14-amino-acid V5 epitope derived from simian parainfluenza virus type 5, was inserted into the C terminus of the capsid protein. Recombinant PCV2 expressing the V5 epitope, recPCV2/CL-V5, was rescued by transfecting an infectious clone into PK-15 cells and was characterised by an immunoperoxidase monolayer assay (IPMA), a serum neutralisation assay (SNA), a capture enzyme-linked immunosorbent assay (ELISA) and immunoelectron microscopy. The V5 epitope was detected in the recombinant marker virus by IPMA and capture ELISA. Furthermore, there was no detectable difference in the antigenicity of the recombinant marker virus compared with the parental virus by IPMA and SNA using PCV2-positive serum and the neutralising monoclonal antibody 1D2. However, recPCV2/CL-V5 marker virus could be differentiated from the parental virus by PCR, IPMA and capture ELISA. The recombinant marker virus was stable on multiplication through 10 passages in PK-15 cells, with a maximum titre of 10(6.25) 50% tissue culture infective dose (TCID(50))/ml. BALB/c mice were inoculated with the recombinant or parental virus via the intranasal and intraperitoneal routes. The parental and recombinant viruses both could replicate in mice, cause microscopic pathological changes, and induce mice to generate anti PCV2 antibodies. Furthermore, the recombinant marker virus could also induce anti V5 epitope tag antibodies. These results indicated that V5 epitope could be displayed on the surface of the capsid protein by inserting its gene just before stop codon of open reading frame 2. More importantly, insertion of the V5 epitope did not seem to interfere with biological characterisation of the recPCV2/CL-V5 marker virus. PMID- 21641945 TI - The community ecology of barley/cereal yellow dwarf viruses in Western US grasslands. AB - Research on plant viruses in natural ecosystems has been increasing rapidly over the past decade. This paper reviews recent research on the barley and cereal yellow dwarf viruses (B/CYDVs) in grasslands of the western US, beginning with the evidence that the disease caused by these viruses facilitated the invasion of western US grasslands by European annual grasses. Observational and experimental studies of B/CYDVs were carried out along a latitudinal gradient (33.8-48.8 degrees N) from southern California to southern Canada. The prevalence and community composition of B/CYDVs were assessed over a variety of scales and under a range of biotic and abiotic conditions. The findings indicate that both biotic and abiotic factors are important influences on virus ecology and epidemiology. Introduced annual grasses are high-quality hosts that amplify both virus and vector populations in this system, but our research suggests that endemic perennial grasses are critically important for sustaining virus populations in contemporary grasslands largely composed of introduced species. Experiments indicated that increased phosphorus supply to hosts resulted in greater host biomass and higher virus prevalence. Using experimental exclosures, it was found that the presence of grazing vertebrate herbivores increased the abundance of annual grasses, resulting in increased virus prevalence. The results of these studies suggest that patterns of B/CYDV prevalence and coinfection in western US grasslands are strongly shaped by the interactions of host plants, vectors, vertebrate herbivores, and abiotic drivers including nutrients. PMID- 21641946 TI - Targeted chemoimmunotherapy using drug-loaded aptamer-dendrimer bioconjugates. AB - We reported an innovative, targeted chemoimmuno drug-delivery system. Although chemoimmunotherapy, as an alternative to or in combination with conventional therapeutic systems, has been in the forefront of recent oncological research, as presently configured, such systems face several major obstacles for efficient clinical application. Here, we establish a novel nano-platform for effective chemoimmunotherapy designed to overcome the drawbacks of conventional cancer therapies, describing a delivery system based on a dendrimer and a single-strand DNA-A9 PSMA (prostate-specific membrane antigen) RNA aptamer hybrid. Employing these vehicles, we demonstrate the promising possibility of this chemoimmuno therapeutic system against prostate cancer in in vivo and in vitro models. PMID- 21641947 TI - Trustworthy satiety claims are good for science and society. Comment on 'Satiety. No way to slim'. AB - In their short communication against satiety claims, Booth and Nouwen (2010) neglect dozens of well designed studies that show consistent relations between satiety, energy intake and body weight. Satiety, intake and weight are separate concepts, that need different claims and evidence to support them. Satiety can be measured reliably. A repeated higher satiety response to a specific food compared to an appropriate control food may be valuable to consumers who want to avoid hunger. This is good for society. The development of the psycho-biological knowledge to achieve this is good for science. The lawmaker should provide the frame of reference for trustworthy satiety claims. It is then up to the consumer to decide the value of these claims. PMID- 21641948 TI - Hyperleptinemia in obese adolescents deregulates neuropeptides during weight loss. AB - Leptin has emerged over the past decade as a key hormone not only in energy balance regulation but also in neuroendocrine and inflammatory processes. The aim of the present study was to evaluate whether hyperleptinemia deregulates neuropeptides during weight loss. A total of 86 post-pubertal obese adolescents (with or without hyperleptinemia) participated in one year of interdisciplinary weight loss therapy (clinical, nutritional, psychological and exercise-related). Adipokine and neuropeptide concentrations were measured by ELISA, visceral fat was measured by ultrasound and body composition was measured by pletismography. The hyperleptinemic patients presented a lower alpha-MSH concentration and higher NPY/AgRP ratio while the adiponectin/leptin (A/L) ratio was lower compared with the non-hyperleptinemic group. After therapy, significant improvements in BM, BMI, body fat mass, visceral and subcutaneous fat, HOMA-IR, QUICKI, total cholesterol and triglycerides were observed in both groups. Indeed, we observed significant increases in adiponectin and A/L as well as reductions in leptin and NPY/AgRP ratio in the hyperleptinemic group. In the stepwise multiple linear regression analysis with leptin concentration as the dependent variable, alpha MSH and body fat mass (%) were the independent predictors to explain leptin concentration. For the entire group, we found positive correlations between leptinemia and BMI and body fat mass (%) as well as a negative correlation with free fat mass (%) and alpha-MSH. Finally, we verified negative correlations between adiponectin/leptin ratio with total cholesterol and LDL-c, only in hyperleptinemic patients. In conclusion, the hyperleptinemia in obese adolescents deregulates neuropeptides during weight loss. PMID- 21641949 TI - Functional variants of the genes involved in neurodevelopment and susceptibility to schizophrenia in an Armenian population. AB - Aberrant neurodevelopment contributes to the etiology of schizophrenia. This study aimed to investigate the potential association of netrin G1 (NTNG1) rs628117 and brain-derived neurotrophic factor (BDNF) Val66Met (rs6265) genetic polymorphisms with susceptibility to schizophrenia. One hundred three Armenian patients with schizophrenia and 105 healthy control subjects were genotyped by polymerase chain reaction with sequence-specific primers. Whereas the NTNG1 rs628117 genotypes were equally distributed in the groups, the carriers of the less common BDNF 66Met allele were overrepresented among patients with schizophrenia when compared with healthy controls (55% vs 35%, odds ratio = 2.28, 95% confidence interval 1.14-1.98, p(corrected) = 0.006). Furthermore, the 66Met/Met genotype correlated with earlier disease onset (p = 0.024). In conclusion, our single-cohort study nominates the BDNF 66Met allele as a risk factor for schizophrenia in an Armenian population. This must be confirmed in other Armenian cohorts. PMID- 21641950 TI - Association of IL10 promoter polymorphisms with idiopathic achalasia. AB - Idiopathic achalasia is an esophageal motor disorder of unknown etiology. A wealth of evidence supports the concept that achalasia is an immune-mediated disease. According to this evidence, achalasia has been significantly associated with specific alleles of the human leukocyte antigen class II, PTPN22 and IL23R. Several studies have demonstrated association of the IL10 gene with different inflammatory disorders. Our aim was to evaluate the role of functional IL10 promoter polymorphisms in susceptibility to idiopathic achalasia. A case-control study was performed with the -1082, -819, and -592 IL10 promoter polymorphisms in 282 patients and 529 controls and in an independent replication set of 75 patients and 575 controls. The GCC haplotype of the IL10 promoter was reported to be associated with a lower risk of achalasia in the discovery sample (odds ratio [OR] = 0.79, 95% confidence interval [CI] = 0.64-0.98, p = 0.029). This association was validated in a replication set (OR = 0.69, 95% CI = 0.48-1.00, p = 0.04). In the combined analysis no heterogeneity was observed between the 2 sample sets and the GCC haplotype was significantly associated with the disease (OR(MH) = 0.76, 95% CI = 0.63-0.91, p = 0.003). Our results provide the first evidence for an association between IL10 promoter polymorphisms and idiopathic achalasia, suggesting that the interleukin-10 cytokine may contribute to the pathogenesis of this disease. PMID- 21641951 TI - Monovalent type 1 oral poliovirus vaccine among infants in India: report of two randomized double-blind controlled clinical trials. AB - BACKGROUND: To provide the polio eradication initiative with more immunogenic oral poliovirus vaccines (OPVs), we evaluated newly developed monovalent type 1 OPV (mOPV1) among infants in India. METHODS: Two double-blind randomized controlled clinical trials compared two mOPV1s (mOPV1 A and mOPV1 B) versus trivalent OPV (tOPV X) given at birth (trial I), or assessed two products of higher-potency mOPV1 (mOPV1 C and mOPV1 D) versus regular-potency mOPV1 (mOPV1 B) or tOPV Y given at birth and at 30 days (trial II). RESULTS: In trial I, 597 newborns were enrolled, 66 withdrawn or excluded, leaving 531 (88.9%) subjects for analysis. Seroconversion to poliovirus type 1 was 10.4% for mOPV1 A, 15.6% for mOPV1 B and 10.2% for tOPV X. In trial II, 718 newborns were enrolled, 135 withdrawn or excluded, leaving 583 (81.2%) subjects for analysis. Seroconversion to poliovirus type 1 following a birth dose was 15.1%, 19.7%, 18.0% and 10.6%, following the 30-day dose 87.1%, 89.2%, 84.4%, or 55.9%, and cumulative for both doses 90.4%, 90.3%, 89.5% and 61.9% for mOPV1s B, C, and D and tOPV Y, respectively. CONCLUSIONS: In both studies, seronconversion rates were unexpectedly low to poliovirus type 1 after mOPV1 or tOPV given at birth but high for all formulations of mOPV1 given at age 30 days. The cause for low immunogenicity of OPV at birth in India is not known. PMID- 21641952 TI - Costs and impact of meningitis epidemics for the public health system in Burkina Faso. AB - BACKGROUND: Epidemic meningococcal meningitis remains a serious health threat in the African meningitis belt. New meningococcal conjugate vaccines are relatively costly and their efficiency will depend on cost savings realized from no longer having to respond to epidemics. METHODS: We evaluated the cost and impacts to the public health system of the 2007 epidemic bacterial meningitis season in Burkina Faso through a survey at the different level of the health system. A micro economic approach was used to evaluate direct medical and non medical costs for both the public health system and households, as well as indirect costs for households. RESULTS: The total national cost was 9.4 million US$ (0.69 US$ per capita). Health system costs were 7.1 million US$ (1.97% of annual national health spending), with 85.6% for reactive vaccination campaigns. The remaining 2.3 million US$ was borne by households of meningitis cases. The mean cost per person vaccinated was 1.45 US$; the mean cost of case management per meningitis case was 116.3 US$ when including household costs and 26.4 US$ when including only health sector costs. Meningitis epidemics disrupted all health services from national to operational levels with the main contributor being a large increase in medical consultations. CONCLUSIONS: Preventive meningococcal conjugate vaccines should contribute to more efficient use of funds dedicated to meningitis epidemics and limit the disruption of routine health services. PMID- 21641953 TI - Vaccine adjuvant activity of 3M-052: an imidazoquinoline designed for local activity without systemic cytokine induction. AB - The human Toll-like receptors (TLRs) are a family of receptors, which sense the presence of various structural elements of pathogens and damaged or effete components in the host. As they do so, they activate two critical arms of host defense, the rapid innate immune response and an adaptive immune response. The innate immune response is typified by the generation of Th1 cytokines, chemokines and type 1 interferons. As such, agonists for the TLRs have potential as antiviral and anticancer therapeutics. They are also well suited to function as vaccine adjuvants. 3M imidazoquinoline (IRM) molecules were the first synthetic small molecules identified as TLR agonists and can affect their biological activities through TLR7, TLR8, or both. The breadth of therapeutic opportunities for this family of molecules can require formulations tailored to the specific application. One consideration is specific formulations to avoid a systemic distribution of these TLR agonists and resulting cytokine storm-like effects on the host. 3M-052 is an IRM bearing a C18 lipid moiety and designed for slow dissemination from the site of application. In the present study 3M-052 has been evaluated for its in vitro TLR activity and for its efficacy as a vaccine adjuvant using a recombinant hemagglutinin from H1N1 A/Puerto Rico/8/34. Given subcutaneously, 3M-052 drives a strong Th1 response to hemagglutinin and serum neutralization of viable H1N1 A/Puerto Rico/8/34 virus in the absence of circulating TNFalpha or the induction of Th1 cytokines. PMID- 21641954 TI - Effectiveness of the whole-cell pertussis vaccine produced in Poland against different Bordetella parapertussis isolates in the mouse intranasal challenge model. AB - The aim of the study was to evaluate the effectiveness of the whole-cell pertussis vaccine produced locally and routinely used in Poland in the elimination of Bordetella parapertussis strains from the lungs and trachea of a mouse model. We found that the average protective effect against B. parapertussis in the lungs of mice immunized with the whole-cell pertussis vaccine (DTwP) was significantly higher than in animals immunized with the acellular pertussis vaccine (DTaP). The effectiveness of B. parapertussis elimination rates from the lungs of DTwP-immunized mice, depending on the strain used as a challenge, was found to be 1.2-3.0 times or 3.1-7.0 times lower than against Bordetella. pertussis Tohama I or vaccine B. pertussis 606/67 isolates, respectively. Our results show that the locally produced DTwP vaccine is able to protect against B. parapertussis isolates; however, the level of protection and course of B. parapertussis infection in the lungs and trachea seems to be strain specific. PMID- 21641955 TI - Nonclinical biodistribution, integration, and toxicology evaluations of an H5N1 pandemic influenza plasmid DNA vaccine formulated with Vaxfectin(r). AB - Vaxfectin((r)) is a lipid-based adjuvant initially developed for use with plasmid DNA (pDNA) vaccines. Here we present detailed nonclinical assessments performed prior to Vaxfectin((r))'s first-in-man use, as an adjuvant in the H5N1 influenza vaccine VCL-IPT1. Following IM delivery to rabbits, VCL-IPT1 pDNA localized primarily to injection sites, where levels steadily declined over the 2 months examined. Risk of pDNA integration into genomic DNA was negligible. Toxicology studies in rabbits revealed mild inflammatory/immune responses at injection sites characteristic of IM vaccine delivery; Vaxfectin((r)) directly contributed to these responses. These data support clinical development of H5N1 pDNA vaccines, and also present an encouraging profile for further development of Vaxfectin((r)) as an adjuvant for vaccines in general. PMID- 21641956 TI - Potential protective immunogenicity of recombinant Clostridium perfringens alpha beta2-beta1 fusion toxin in mice, sows and cows. AB - Clostridial toxins are main pathogenic virulence of Clostridium perfringens that have been associated with a wide range of diseases in both humans and domestic animals. Genetically engineered toxoids have been shown to function as potential vaccine candidates in the prevention of Clostridium derived infectious diseases. In this study, we have developed recombinant alpha-toxin (CPA), beta2/beta1 fusion toxin (CPB2B1) and alpha/beta2/beta1 trivalent fusion-toxin (CPAB2B1) as vaccine candidates that may be used to vaccinate against C. perfringens alpha, beta1 and beta2-toxins. Mice immunized with these recombinant toxoids demonstrated a strong protective immunological response when administered a lethal dose of C. perfringens type C culture filtrate with high titers of neutralizing antibodies to the toxins in the sera, as well as the intestinal mucosal s-IgA level. Specific neutralizing antibodies to the toxins were also detected in the sera and colostrum of sows and cows vaccinated with the toxoids. Furthermore, the CPA and CPB2B1 recombinant toxoid cocktail was capable of stimulating relatively higher levels of immune responses compared to that of CPA, CPB2B1 and CPAB2B1 alone. The CPAB2B1 trivalent fusion toxoid also displayed increased immunogenicity relative to CPA and CPB2B1 alone. These results suggest that recombinant toxoids are potential vaccine candidates against Clostridial toxins; the use of mixed cocktails and/or multivalent recombinant toxoids against different types of toxins may be an effective approach in the prevention of diseases caused by toxins produced by C. perfringens. PMID- 21641957 TI - A single sip of a strong alcoholic beverage causes exposure to carcinogenic concentrations of acetaldehyde in the oral cavity. AB - The aim of this study was to explore oral exposure to carcinogenic (group 1) acetaldehyde after single sips of strong alcoholic beverages containing no or high concentrations of acetaldehyde. Eight volunteers tasted 5 ml of ethanol diluted to 40 vol.% with no acetaldehyde and 40 vol.% calvados containing 2400 MUM acetaldehyde. Salivary acetaldehyde and ethanol concentrations were measured by gas chromatography. The protocol was repeated after ingestion of ethanol (0.5 g/kg body weight). Salivary acetaldehyde concentration was significantly higher after sipping calvados than after sipping ethanol at 30s both with (215 vs. 128 MUmol/l, p<0.05) and without (258 vs. 89 MUmol/l, p<0.05) alcohol ingestion. From 2 min onwards there were no significant differences in the decreasing salivary acetaldehyde concentration, which remained above the level of carcinogenicity still at 10 min. The systemic alcohol distribution from blood to saliva had no additional effect on salivary acetaldehyde after sipping of the alcoholic beverages. Carcinogenic concentrations of acetaldehyde are produced from ethanol in the oral cavity instantly after a small sip of strong alcoholic beverage, and the exposure continues for at least 10 min. Acetaldehyde present in the beverage has a short-term effect on total acetaldehyde exposure. PMID- 21641958 TI - Localization of MMP-2, MMP-9, TIMP-1, and TIMP-2 in human coronal dentine. AB - OBJECTIVES: Matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) play important roles in dentine formation, caries progression and hybrid layer degradation. This study tested the hypothesis that the distribution and concentrations of MMP-2, MMP-9, TIMP-1 and TIMP-2 are different at different depths of human coronal dentine, including odontoblasts. METHODS: Protein localization was performed using immunohistochemistry. Co-localization of the MMPs and their inhibitors was conducted using immunofluorescence double labelling. Protein concentrations were measured by ELISA and gelatinolytic potential was assessed with gelatine zymography. RESULTS: MMP-2 was the main gelatinase in dentine and was concentrated in the odontoblasts, deep dentine and the dentinoenamel junction. TIMP-2 was co-localized with MMP-2 mainly in the odontoblasts but its concentration was low. Both MMP-9 and TIMP-1 showed a decreasing distribution from the deep to the superficial dentine layers; however, the concentration of TIMP-1 was much higher than that of MMP-9. The gelatinolytic potential of dentine protein extracts decreased gradually from deep to superficial dentine. CONCLUSIONS: The concentrations and distribution patterns of MMP-2, MMP-9, TIMP-1 and TIMP-2, and the gelatinolytic potential of dentine matrix are variable along different dentine depths. Thus, differential collagen degradation potentials may be expected depending upon the depth in which dentine is exposed. PMID- 21641959 TI - Surface area accessibility and the preferred topology of telomeric DNA quadruplex ligand complexes. AB - Solvent-accessible surface area calculations have been performed on two human telomeric quadruplex structures, the parallel crystal structure and an (3 + 1) anti-parallel structure determined by NMR methods. The differences in net ligand solvent-accessible surface area (Delta(SASA)) for four structurally distinct categories of small-molecule ligand have been computed, using docked structures of complexes with both types of quadruplex, as well as the relative contributions of polar and non-polar surface areas. It has been hypothesized that the surface area occupied by the ligand is a determinant of selectivity between different quadruplex topologies, where the ligand maximizes the accessible surface area by contacting all accessible atoms at one end of a quadruplex structure. This has enabled selectivity for a particular ligand to be assessed, for parallel compared an anti-parallel topology. The predictions for the ligands chosen, all of which have their quadruplex topological preferences experimentally determined and reported in the literature, are fully in accord with observation. It is suggested that this approach, which does not depend on energy functions, can be useful in the rational design of topology-specific ligands, especially in the case of polymorphic quadruplexes. PMID- 21641960 TI - The triazatruxene derivative azatrux binds to the parallel form of the human telomeric G-quadruplex under molecular crowding conditions: biophysical and molecular modeling studies. AB - The present study has employed a combination of spectroscopic, calorimetric and computational methods to explore the binding of the three side-chained triazatruxene derivative, termed azatrux, to a human telomeric G-quadruplex sequence, under conditions of molecular crowding. The binding of azatrux to the tetramolecular parallel [d(TGGGGT)](4) quadruplex in the presence and absence of crowding conditions, was also characterized. The data indicate that azatrux binds in an end-stacking mode to the parallel G-quadruplex scaffold and highlights the key structural elements involved in the binding. The selectivity of azatrux for the human telomeric G-quadruplex relative to another biologically relevant G quadruplex (c-Kit87up) and to duplex DNA was also investigated under molecular crowding conditions, showing that azatrux has good selectivity for the human telomeric G-quadruplex over the other investigated DNA structures. PMID- 21641961 TI - Fluorescence intercalator displacement assay for screening G4 ligands towards a variety of G-quadruplex structures. AB - The potential formation of G-quadruplexes in many regions of the genome makes them an attractive target for drug design. A large number of small molecules synthesized in recent years display an ability to selectively target and stabilize G-quadruplexes. To screen for G4 ligands, we modified a G4-FID (G quadruplex Fluorescent Intercalator Displacement) assay. This test is based on the displacement of an "on/off" fluorescence probe, Thiazole Orange (TO), from quadruplex or duplex DNA matrices by increasing amounts of a putative ligand. Selectivity measurements can easily be achieved by comparing the ability of the ligand to displace TO from various quadruplex and duplex structures. G4-FID requires neither modified oligonucleotides nor specific equipment and is an isothermal experiment. This test was adapted for high throughput screening onto 96-well plates allowing the comparison of more than twenty different structures. Fifteen different known G4 ligands belonging to different families were tested. Most compounds showed a good G4 vs duplex selectivity but exhibited little, if any, specificity for one quadruplex sequence over the others. The quest for the "perfect" specific G4 ligand is not over yet! PMID- 21641962 TI - Bax: Addressed to kill. AB - The pro-apoptototic protein Bax (Bcl-2 Associated protein X) plays a central role in the mitochondria-dependent apoptotic pathway. In healthy mammalian cells, Bax is essentially cytosolic and inactive. Following a death signal, the protein is translocated to the outer mitochondrial membrane, where it promotes a permeabilization that favors the release of different apoptogenic factors, such as cytochrome c. The regulation of Bax translocation is associated to conformational changes that are under the control of different factors. The evidences showing the involvement of different Bax domains in its mitochondrial localization are presented. The interactions between Bax and its different partners are described in relation to their ability to promote (or prevent) Bax conformational changes leading to mitochondrial addressing and to the acquisition of the capacity to permeabilize the outer mitochondrial membrane. PMID- 21641963 TI - Development of hydrocephalus and classical hypothesis of cerebrospinal fluid hydrodynamics: facts and illusions. AB - According to the classical hypothesis of the cerebrospinal fluid (CSF) hydrodynamics, CSF is produced inside the brain ventricles, than it circulates like a slow river toward the cortical subarachnoid space, and finally it is absorbed into the venous sinuses. Some pathological conditions, primarily hydrocephalus, have also been interpreted based on this hypothesis. The development of hydrocephalus is explained as an imbalance between CSF formation and absorption, where more CSF is formed than is absorbed, which results in an abnormal increase in the CSF volume inside the cranial CSF spaces. It is believed that the reason for the imbalance is the obstruction of the CSF pathways between the site of CSF formation and the site of its absorption, which diminishes or prevents CSF outflow from the cranium. In spite of the general acceptance of the classical hypothesis, there are a considerable number of experimental results that do not support such a hypothesis and the generally accepted pathophysiology of hydrocephalus. A recently proposed new working hypothesis suggests that osmotic and hydrostatic forces at the central nervous system microvessels are crucial for the regulation of interstial fluid and CSF volume which constitute a functional unit. Based on that hypothesis, the generally accepted mechanisms of hydrocephalus development are not plausible. Therefore, the recent understanding of the correlation between CSF physiology and the development of hydrocephalus has been thoroughly presented, analyzed and evaluated, and new insights into hydrocephalus etiopathology have been proposed, which are in accordance with the experimental data and the new working hypothesis. PMID- 21641964 TI - Personality-dependent effects of oxytocin: greater social benefits for high alexithymia scorers. AB - Originally known for its role in labor and lactation, oxytocin (OT) has recently been shown to facilitate social behaviour by improving socio-emotional abilities. However, whether OT is equally beneficial to all people, or whether is it particularly beneficial to less emotionally/socially competent (i.e., high alexithymia) individuals it is not yet known. We investigated the effects of OT on individuals of varying socio-emotional ability by randomly assigning sixty male students to receive either oxytocin (OT) or a placebo (PL), and had them perform the Reading the Mind in the Eyes test (RMET). Results showed that whereas the performance of lower alexithymia individuals was equally good in both OT and PL conditions, the performance of higher alexithymia people was better under OT than PL. These results suggest that the effects of OT are not only context dependent-as recently shown-but also personality dependent. They also provide new insights into the remediation of socio-emotional deficits. PMID- 21641965 TI - Opposing roles for E2F1 in survival and death of cerebellar granule neurons. AB - The transcription factor E2F1 is upregulated when cerebellar granular neurons (CGNs) undergo apoptosis under potassium deprivation. In this study, we examined the effects of E2F1 upregulation on the survival and death of CGNs isolated from C57 mice and Sprague-Dawley (SD) rats. Plasmid- and adenovirus-mediated expression of E2F1 dose-dependently induced apoptosis in mouse CGNs but unexpectedly failed to induce apoptosis in rat CGNs. Caspase 3, a marker for neuronal apoptosis, was significantly activated by ectopic E2F1 expression in mouse CGNs but not in rat CGNs. Furthermore, overexpression of E2F1 significantly promoted apoptotic progression in mouse CGNs following potassium deprivation but attenuated apoptosis in rat CGNs, whereas E2F1 lacking DNA binding ability (E2F1 M132) lost its pro-apoptotic role in mouse CGNs and anti-apoptotic role in rat CGNs. Together, our results demonstrated that upregulation of E2F1 by potassium deprivation promotes apoptosis in C57 mouse CGNs but antagonizes apoptosis in SD rat CGNs, suggesting opposing roles for E2F1 in regulating CGN fate. PMID- 21641966 TI - Broadband dielectric spectroscopy on human blood. AB - BACKGROUND: Dielectric spectra of human blood reveal a rich variety of dynamic processes. Achieving a better characterization and understanding of these processes not only is of academic interest but also of high relevance for medical applications as, e.g., the determination of absorption rates of electromagnetic radiation by the human body. METHODS: The dielectric properties of human blood are studied using broadband dielectric spectroscopy, systematically investigating the dependence on temperature and hematocrit value. By covering a frequency range from 1Hz to 40GHz, information on all the typical dispersion regions of biological matter is obtained. RESULTS AND CONCLUSIONS: We find no evidence for a low-frequency relaxation ("alpha-relaxation") caused, e.g., by counterion diffusion effects as reported for some types of biological matter. The analysis of a strong Maxwell-Wagner relaxation arising from the polarization of the cell membranes in the 1-100MHz region ("beta-relaxation") allows for the test of model predictions and the determination of various intrinsic cell properties. In the microwave region beyond 1GHz, the reorientational motion of water molecules in the blood plasma leads to another relaxation feature ("gamma-relaxation"). Between beta- and gamma-relaxations, significant dispersion is observed, which, however, can be explained by a superposition of these relaxation processes and is not due to an additional "delta-relaxation" often found in biological matter. GENERAL SIGNIFICANCE: Our measurements provide dielectric data on human blood of so far unsurpassed precision for a broad parameter range. All data are provided in electronic form to serve as basis for the calculation of the absorption rate of electromagnetic radiation and other medical purposes. Moreover, by investigating an exceptionally broad frequency range, valuable new information on the dynamic processes in blood is obtained. PMID- 21641967 TI - Genetic susceptibility to sporadic ovarian cancer: a systematic review. AB - Ovarian cancer is a highly lethal disease. Many researchers have, therefore, attempted to identify high risk populations. In this perspective, numerous genetic association studies have been performed to discover common ovarian cancer susceptibility variants. Accordingly, there is an increasing need to synthesize the evidence in order to identify true associations. A comprehensive and systematic assessment of all available data on genetic susceptibility to sporadic ovarian cancer was carried out. The evidence of statistically significant findings was evaluated based on the number of positive replications, the ratio of positive and negative studies, and the false-positive report probability (FPRP). The authors reviewed three genome-wide association studies (GWAS) and 147 candidate gene studies, published from 1990 to October 2010, including around 1100 genetic variants in more than 200 candidate genes and 20 intergenic regions. Genetic variants with the strongest evidence for an association with ovarian cancer include the rs2854344 in the RB1 gene and SNPs on chromosomes 9p22.2, 8q24, 2q31, and 19p13. Promising genetic pathways for ovarian cancer include the cell cycle, DNA repair, sex steroid hormone and oncogenic pathway. Concluding, this review shows that many genetic association studies have been performed, but only a few genetic variants show strong evidence for an association with ovarian cancer. More research is needed to elucidate causal genetic variants, taking into consideration gene-gene and gene-environment interactions, combined effects of common and rare variants, and differences between histological subtypes of this cancer. PMID- 21641968 TI - The discovery of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor complex and the molecular regulation of synaptic vesicle transmitter release: the 2010 Kavli Prize in neuroscience. AB - Brain function depends on a crucial feature: The ability of individual neurons to share packets of information, known as quantal transmission. Given the sheer number of tasks the brain has to deal with, this information sharing must be extremely rapid. Synapses are specialized points of contact between neurons, where fast transmission takes place. Though the basic elements and functions of the synapse had been established since the 1950s, the molecular basis for regulation of fast synaptic transmitter release was not known 20 years ago. However, around 1990, crucial discoveries were made by Richard Scheller, James Rothman, and Thomas Sudhof, leading a few years later to the formulation of the SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) hypothesis and a new understanding of the molecular events controlling vesicular release of transmitter in synapses. The 2010 Kavli Prize in neuroscience was awarded to these three researchers, "for their work to reveal the precise molecular basis of the transfer of signals between nerve cells in the brain." PMID- 21641969 TI - Neuroactive steroids in periphery and cerebrospinal fluid. AB - Some peripheral steroids penetrate the blood-brain barrier (BBB), providing at least substances for the CNS steroid metabolome. That is why the predictive value of the peripheral steroids appears to be comparable with that of the cerebrospinal fluid (CSF) steroids. The concentrations of the CSF steroids are pronouncedly lower in comparison with the ones in circulation. The available data indicate that the levels of pregnenolone sulfate substantially increase in the rat brain tissue after the administration of pregnenolone into the circulation. In the human circulation there are about two orders of magnitude higher levels of pregnenolone sulfate compared to the free pregnenolone. Our data show insignificant correlation between CSF and serum pregnenolone, but a borderline one between CSF pregnenolone and serum pregnenolone sulfate. Therefore in humans, the circulating pregnenolone sulfate might be of an importance for pregnenolone concentration in the CNS. In contrast to free pregnenolone, dehydroepiandrosterone (DHEA) in the CSF correlates with both unconjugated and conjugated DHEA in the serum. These data as well as the low C17-hydroxylase C17,20-lyase activity in the CNS might indicate that DHEA levels in the CNS are influenced by peripheral levels of DHEA and its sulfate. According to the information, available part of the neurosteroids may be synthesized de novo in the CNS, but substantial part of the steroid metabolites may be also synthesized in the CNS from the steroid precursors or directly transported through BBB from the periphery. The processes mentioned above may be complimentary in some cases. Brain synthesis may provide minimal level of neurosteroids, which are indispensable for the CNS functions. Thus, brain steroids of peripheral origin may reflect various physiological situations or even pathologies. This article is part of a Special Issue entitled: Neuroactive Steroids: Focus on Human Brain. PMID- 21641970 TI - Berberine hydrochloride attenuates cyclooxygenase-2 expression in rat small intestinal mucosa during acute endotoxemia. AB - The effect of berberine hydrochloride (BBR) on inducible cyclooxygenase-2 (COX-2) in small intestinal mucosa and related mechanisms was investigated in a rat model of acute endotoxemia. The results showed that lipopolysaccharide (LPS) increased COX-2 expression, whereas SB202190 and BBR curtailed it. LPS increased phosphorylation of mucosal p38 MAPK and ATF2 as well as production of ATF2, whereas BBR attenuated these effects. LPS upregulated mucosal peroxisome proliferator-activated receptor gamma (PPARgamma), but BBR reduced this receptor. GW9662 aggravated LPS-induced and reversed BBR-attenuated COX-2 expression. The findings showed that BBR ameliorated COX-2 overexpression partially via modulation of p38 and PPARgamma pathways during acute endotoxemia. PMID- 21641971 TI - New cassane diterpenes from Caesalpinia echinata. AB - An investigation of the ethanolic extract from stems of Caesalpinia echinata Lam (Leguminosae-Caesalpinioideae) led to the isolation of five new cassane diterpenes along with known lambertianic acid. Their structures were determined based on spectroscopic methods. A preliminary study on leishmanicidal activity demonstrated that compounds 1, 2 and 6 were found to inhibit the growth of amastigote-like forms of Leishmania amazonensis without affecting mononuclear cells obtained from human peripheral blood. PMID- 21641972 TI - Cytotoxic alkaloids from stems, leaves and twigs of Dasymaschalon blumei. AB - Bioassay-guided fractionation of the cytotoxic ethyl acetate extract from the stems of Dasymaschalon blumei (Annonaceae) led to the isolation of four aristololactam alkaloids, including the hitherto unknown 3,5-dihydroxy-2,4 dimethoxyaristolactam (1), as well as the three known compounds, aristolactam BI, goniopedaline, and griffithinam. Additionally, the cytotoxic extract from the combined leaves and twigs of the same plant yielded three known oxoaporphine alkaloids, oxodiscoguattine, dicentrinone, and duguevalline. The structures of aristolactams and oxoaporphine alkaloids were elucidated on the basis of spectroscopic methods. All isolates were evaluated for cytotoxicity against a panel of mammalian cancer cell lines and a noncancerous human embryonic kidney cell Hek 293. PMID- 21641973 TI - Sesquiterpene lactones from Saussurea involucrata. AB - Bioassay-directed separation of the ethyl acetate extract from the aerial parts of Saussurea involucrata (Kar. et Kir. ex Maxim) led to the isolation of three new sesquiterpene lactones sausinlactones A (1), B (2), C (3) and six known ones (4-9). The structures were established by spectroscopic data (UV, IR, HRESIMS, 1D and 2D NMR). CD technique was also employed to determine the absolute configurations of new compounds 1-3. The anti-inflammatory activities of compounds 1-9 and antitumor activities of new compounds 1-3 were tested. The results presented that compounds 5 and 6 were responsible for anti-inflammatory activities, and compounds 1 and 2 showed significant cytotoxic activities against A549 cells. PMID- 21641974 TI - Sporophytic and gametophytic functions of the cell cycle-associated Mob1 gene in Arabidopsis thaliana L. AB - Mob1 genes are primarily involved in the cell cycle progression and mitosis exit in yeasts and animals. The function of a Mob1-like gene (At5g45550) from Arabidopsis thaliana was investigated using RNAi and immunological staining. AtMob1-like RNAi silenced lines showed a reduced radial expansion of the inflorescence stem and a reduced elongation zone of the primary root. Morphological features of plant organs were accompanied by a reduction in cell size. The fertility of AtMob1-like RNAi silenced lines was very low as seed production was strongly reduced. About 2% of the progeny of AtMob1-like RNAi silenced plants were tetraploid. The female and male sporogenesis was affected differentially. The ovules developed irregularly and one third of the megaspores and embryo sacs degenerated prematurely. Up to 20% of the ovules produced binucleated megaspores that failed to develop further, being their degeneration likely accompanied with a delayed programmed cell death. The anthers produced about 30% of aborted pollen grains, showing also a strong variation in their size. Together, the results show that Arabidopsis MOB1-like is required to regulate cell expansion and cell division, presumably by affecting the mitotic as well as the meiotic cell cycle. PMID- 21641975 TI - Single primer amplification reaction (SPAR) reveals inter- and intra-specific natural genetic variation in five species of Cymbidium (Orchidaceae). AB - A total of 53 primers belonging to three SPAR methods, viz. RAPD, ISSR and DAMD, collectively produced 456 polymorphic amplicons with 96.6% polymorphism at inter specific level in five species of Cymbidium, viz. C. aloifolium, C. mastersii, C. elegans, C. eburneum and C. tigrinum, whereas at intra-specific level, the observed polymorphism ranged from 51.2% to 77.1% among them. Three SPARs collectively revealed 25 unique species-specific amplicons; most of them were amplified with RAPD and DAMD primers besides few bands which were either missed (absent) or lost (heterozygosity). UPGMA clustering evidently distinguished the representatives of C. aloifolium and C. tigrinum, with distinct genetic distance, which may be due to their entirely different habitats as well as discrete morphological characteristics. Upon analysis of the data generated, all the three SPAR methods, either independently and/or in combination, revealed wide range of genetic variation between and within five species of Cymbidium. Comparison of matrix of individual SPAR method revealed that analysis of natural genetic variation using combination of SPAR methods, rather than an isolated approach, is highly effective. The critical analyses of the amplicon data are indicative of DAMD as the most powerful SPAR method by showing highest resolving power (Rp) followed by ISSR and RAPD. Alternatively, the total polymorphic information content was highest in case of RAPD followed by other two SPAR methods. Thus, the present investigation for the first time provides a valuable baseline data for genetic variation at inter- and intra-specific levels in horticultural Cymbidiums and also addresses conservation concerns. PMID- 21641976 TI - Perfluorinated compounds differentially affect steroidogenesis and viability in the human adrenocortical carcinoma (H295R) in vitro cell assay. AB - Perfluorinated compounds (PFCs) comprise a large class of man-made chemicals of which some are persistent and present throughout the ecosystem. This raises concerns about potential harmful effects of such PFCs on humans and the environment. In order to investigate the effects of potentially harmful PFCs on steroid hormone production, human adrenocortical H295R cells were exposed to three persistent PFCs including perfluorooctane sulfonate (PFOS), perfluorooctanoic acid (PFOA) and perfluorononanoic acid (PFNA) at six different concentrations (6nM to 600MUM) for 48h. Exposure to 600MUM PFOS resulted in a dose-responsive increase in oestradiol as well as a smaller dose-responsive increase in progesterone and testosterone secretion measured using radioimmunoassay. The aromatase activity was not significantly altered by PFOS. Only small changes in hormone secretion were detected following exposure to PFOA and PFNA. Gene expression of CYP11A, quantified using qRT-PCR was decreased by all exposure doses of PFOA, whereas HMGR expression was decreased by 60nM PFNA. The viability markedly decreased by exposure to 600MUM of PFOA or PFNA, but not PFOS. Flow cytometric analysis demonstrated a significant increase in apoptosis following exposure to PFNA at the highest concentration. We conclude that PFOS is capable of altering steroidogenesis in the H295R in vitro model by a mechanism other than changes in gene expression or activity of aromatase. Additionally, PFCs appear to differentially affect cell viability with induction of cell death via apoptosis at high doses of PFNA. PMID- 21641977 TI - Development and application of a physiologically based pharmacokinetic model for triadimefon and its metabolite triadimenol in rats and humans. AB - A physiologically based pharmacokinetic (PBPK) model was developed for the conazole fungicide triadimefon and its primary metabolite, triadimenol. Rat tissue:blood partition coefficients and metabolic constants were measured in vitro for both compounds. Pharmacokinetic data for parent and metabolite were collected from several tissues after intravenous administration of triadimefon to male Sprague-Dawley rats. The model adequately simulated peak blood and tissue concentrations but predicted more rapid clearance of both triadimefon and triadimenol from blood and tissues. Reverse metabolism of triadimenol to triadimefon in the liver was explored as a possible explanation of this slow clearance, with significant improvement in model prediction. The amended model was extrapolated to humans using in vitro metabolic constants measured in human hepatic microsomes. Human equivalent doses (HEDs) were calculated for a rat no observable adverse effect level (NOAEL) dose of 3.4mg/kg/day using area under the concentration curve (AUC) in brain and blood for triadimefon and triadimenol as dosimetrics. All dosimetric-based HEDs were 25-30 fold above the human oral reference dose of 0.03mg triadimefon/kg/day, but did not account for intra-human variability or pharmacodynamic differences. Ultimately, derivations of this model will be able to better predict the exposure profile of these and other conazole fungicides in humans. PMID- 21641978 TI - Chromium(VI) causes down regulation of biotinidase in human bronchial epithelial cells by modifications of histone acetylation. AB - Hexavalent chromium (Cr(VI)), a commonly used industrial metal, is a well-known mutagen and carcinogen, and occupational exposure can induce a broad spectrum of adverse health effects, including cancers. Although Cr(VI)-induced DNA damage is thought to be the primary mechanism of chromate genotoxicity and mutagenicity, there is an increasing number of reports showing that epigenetic mechanisms of gene regulation might be a central target of Cr(VI) toxicity. Epigenetic changes, such as changes in phosphorylation, altered DNA methylation status, histone acetylation and signaling pathways, have been observed after chromium exposure. Nevertheless, to better demonstrate the roles of epigenetic modifications in Cr(VI)-induced carcinogenesis, more work needs to be carried out. This study is aimed to investigate changes in biotinidase (BTD) and holocarboxylase synthetase (HCS), two major proteins which maintain homeostasis of the newfound epigenetic modification: histone biotinylation, in cells exposed to Cr(VI). The data showed that Cr(VI) decreased BTD expression at the transcriptional level in human bronchial epithelial cells (16HBE). In addition, using the epigenetic modifiers, 5-Aza-2'-deoxycytidine (Aza) and Trichostatin A (TSA), we found that modifications of histone acetylation reversed the inhibition of BTD, suggesting that Cr(VI) may cause down regulation of BTD by modifications of histone acetylation. PMID- 21641979 TI - Protection against nerve agent poisoning by a noncompetitive nicotinic antagonist. AB - The acute toxicity of organophosphorus (OP) nerve agents arises from accumulation of acetylcholine (ACh) and overstimulation of ACh receptors. The mainstay of current pharmacotherapy is the competitive muscarinic antagonist, atropine. Nicotinic antagonists have not been used due to the difficulties of administering a dose of a competitive neuromuscular blocker sufficient to antagonise the effects of excessive ACh, but not so much that it paralyses the muscles. An alternative approach would be to use a noncompetitive antagonist whose effects would not be overcome by increasing ACh concentrations. This study demonstrates that the compound 1,1'-(propane-1,3-diyl)bis(4-tert-butylpyridinium), which blocks open nicotinic ion channels noncompetitively, is able to reverse the neuromuscular paralysis after nerve agent poisoning in vitro and to protect guinea pigs against poisoning by nerve agents when used as part of a therapeutic drug combination including a muscarinic antagonist. In contrast to the oxime HI 6, this compound was equally effective in protecting against poisoning by sarin or tabun. Further studies should identify more effective compounds with this action and optimise doses for protection against nerve agent poisoning in vivo. PMID- 21641980 TI - Effects of DMSA-coated Fe3O4 magnetic nanoparticles on global gene expression of mouse macrophage RAW264.7 cells. AB - Fe(3)O(4) magnetic nanoparticles (MNPs) coated with 2,3-dimercaptosuccinnic acid (DMSA) are considered to be a promising nanomaterial with biocompatibility. In the present study, the effects of DMSA-coated Fe(3)O(4) MNPs on the expression of all identified mouse genes, which regulate various cellular biological processes, were determined to establish whether this nanoparticle is cytotoxic to mammalian cells. Mouse macrophage RAW264.7 cells were treated with 100MUg/ml of DMSA-coated Fe(3)O(4) MNPs for 4, 24 and 48h, and the global gene expression was detected via Affymetrix Mouse Genome 430 2.0 GeneChips((r)) microarrays. It was found that gene expression of 711, 545 and 434 transcripts was significantly altered by 4-, 24- and 48-h treatments, respectively. Of these genes, 27 were consistently upregulated and 6 were consistently downregulated at the three treatment durations. Bioinformatic analysis of all differentially expressed genes revealed that this nanoparticle can strongly activate inflammatory and immune responses and can inhibit the biosynthesis and metabolism of RAW264.7 cells at a dose of 100MUg/ml. These results demonstrated that DMSA-coated Fe(3)O(4) MNPs display cytotoxicity in this type of macrophage at high doses. PMID- 21641981 TI - Aflatoxins upregulate CYP3A4 mRNA expression in a process that involves the PXR transcription factor. AB - Pregnane X receptor (PXR) is a member of the nuclear hormone receptor (NHR) superfamily, which regulates xenobiotic and endobiotic metabolism in the liver. This transcription factor is activated by structurally diverse ligands, including drugs and environmental pollutants. PXR regulates the expression of numerous genes that function in biotransformation and the disposition of xenobiotics upon binding to an AG(G/T)TCA DNA motif in target promoter regions. We performed a screen of mycotoxins that pose a known environmental threat to human and animal health for the ability to activate PXR function in a human hepatocyte cell line, HepG2. We found that aflatoxins B1, M1, and G1 activated PXR. This activation was associated with upregulation of CYP3A4 expression and increased occupancy of PXR protein on the CYP3A4 promoter. Using a microarray approach, we also found that aflatoxin B1 upregulated the expression of multiple genes involved in xenobiotic metabolism, including genes known to be regulated in a PXR-dependent fashion. We also observed an effect of aflatoxin B1 on the expression in other functional groups of genes, including the downregulation of genes involved in cholesterologenesis. The results of this study indicate that aflatoxin B1 is able to activate PXR, a known regulator of liver xenobiotic metabolism, in human hepatocytes, and it can upregulate the expression of PXR-dependent genes responsible for aflatoxin B1 biotransformation, including CYP3A4. PMID- 21641982 TI - Liposome preparation using a hollow fiber membrane contactor--application to spironolactone encapsulation. AB - In this study, we present a novel liposome preparation technique suitable for the entrapment of pharmaceutical and cosmetic agents. This new method uses a membrane contactor in a hollow fiber configuration. In order to investigate the process, key parameters influence on the liposome characteristics was studied. It has been established that the vesicle size distribution decreased with the organic phase pressure decrease, the phospholipid concentration decreases and the aqueous to organic phase volume ratio increases. Liposomes were filled with a hydrophobic drug model, spironolactone that could be used for a paediatric medication. The mean size of drug-free and drug-loaded liposomes was, respectively, 113 +/- 4 nm and 123 +/- 3 nm. The zeta potential of drug-free and drug-loaded liposomes was, respectively, -43 +/- 0.7 mV and -23 +/- 0.6 mV. High entrapment efficiency values were successfully achieved (93 +/- 1.12%). Transmission electron microscopy images revealed nanometric sized and spherical shaped oligo-lamellar vesicles. The release profile showed a rapid and complete release within about 5h. Additionally, special attention was paid on process reproducibility and long term lipid vesicles stability. Results confirmed the robustness of the hollow fiber module based technique. Moreover, the technique is simple, fast and has a potential for continuous production of nanosized liposome suspensions at large scale. PMID- 21641983 TI - Treatment of glioblastoma with poly(isohexyl cyanoacrylate) nanoparticles. AB - Glioblastomas belong to the most devastating cancer diseases. For this reason, polysorbate 80 (Tween 80)-coated poly(isohexyl cyanoacrylate) (PIHCA) (Monorex) nanoparticles loaded with doxorubicin were developed and tested for their use for the treatment of glioblastomas. The preparation of the nanoparticles resulted in spherical particles with high doxorubicin loading. The physico-chemical properties and the release of doxorubicin from the PIHCA-nanoparticles were analysed, and the influence on cell viability of the rat glioblastoma 101/8-cell line was investigated. In vitro, the empty nanoparticles did not show any toxicity, and the anti-cancer effects of the drug-loaded nanoparticles were increased in comparison to doxorubicin solution, represented by IC(50) values. The in vivo efficacy was then tested in intracranially glioblastoma 101/8-bearing rats. Rats were treated with 3 * 1.5mg/kg doxorubicin and were sacrificed 18 days after tumour transplantation. Histological and immunohistochemical analyses were carried out to assess the efficacy of the nanoparticles. Tumour size, proliferation activity, vessel density, necrotic areas, and expression of glial fibrillary acidic protein demonstrated that doxorubicin-loaded PIHCA nanoparticles were much more efficient than the free drug. The results suggest that poly(isohexyl cyanoacrylate) nanoparticles hold great promise for the non invasive therapy of human glioblastomas. PMID- 21641984 TI - In vitro and in vivo evaluation of an intraocular implant for glaucoma treatment. AB - Implantable disks for glaucoma treatment were prepared by blending poly(E caprolactone), PCL, poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide) and dorzolamide. Their in vivo performance was assessed by their capacity to decrease intraocular pressure (IOP) in normotensive and hypertensive eyes. Drug mapping showed that release was complete from blend disks and the low molecular weight (MW) PCL after 1 month in vivo. The high MW PCL showed non cumulative release rates above the therapeutic level during 3 months in vitro. In vivo, the fibrous capsule formation around the implant controls the drug release, working as a barrier membrane. Histologic analysis showed normal foreign body reaction response to the implants. In normotensive eyes, a 20% decrease in IOP obtained with the disks during 1 month was similar to Trusopt eyedrops treatment. In hypertensive eyes, the most sustained decrease was shown by the high MW PCL (40% after 1 month, 30% after 2 months). It was shown that the implants can lower IOP in sustained manner in a rabbit glaucoma model. PMID- 21641985 TI - Preparation and characterization of pH-independent sustained release tablet containing solid dispersion granules of a poorly water-soluble drug. AB - Sustained release (SR) tablets containing solid dispersions (SD) granules of a poorly water-soluble drug were prepared to investigate the controlled pH independent release of the drug. Losartan potassium (LST), an anti-hypertensive agent was chosen as a model drug because of its pH-dependent solubility and short elimination half-life. Poloxamer 188 was used as an SD carrier. A free-flowing SD granule was prepared by adsorbing the melt of the drug and poloxamer 188 onto the surface of an adsorbent, Aerosil 300 (fumed silicon dioxide), followed by direct compression with polyethylene oxide (PEO, 5 * 10(6)) to obtain an SD-loaded SR (SD-SR) matrix tablet. Differential scanning calorimetry (DSC) and powder X-ray diffraction (PXRD) revealed partially amorphous structures of the drug in the SD granules. Scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS) images indicated adsorption of SD granules onto the surface of the adsorbent. The SD granules dissolved completely within 10 min, a dissolution rate much higher than that of pure LST. Moreover, pH-independent sustained release of LST from the SD-SR tablet was achieved for 2h in gastric fluid (pH 1.2) and for 10h in intestinal fluid (pH 6.8). A combination of SD techniques using surface adsorption and SR concepts is a promising approach to control the release rate of poorly water-soluble drugs in a pH-independent manner. PMID- 21641986 TI - Dual imaging of mRNA and protein production: an investigation of the mechanism of heterogeneity in cationic lipid-mediated transgene expression. AB - Heterogeneity of transgene expression is a severe disadvantage in the use of cationic lipid-mediated gene vectors. We previously demonstrated that heterogeneity of the post-nuclear delivery process, as well as intracellular trafficking (i.e. nuclear delivery) is a major determinant in the overall heterogeneity in gene expression, when plasmid DNA (pDNA) is transfected to HeLa cells using a lipoplex. In this study, we explored the mechanism underlying this heterogeneity in a post-nuclear transport process by the dual imaging of mRNA and its encoded protein (histone H2B-tagged mTFP1; mTFP1-H2B) in a single cell. To establish a highly sensitive imaging system for mRNA, we used fluorescence in situ hybridization (FISH) combined with tyramide signal amplification (TSA) and a semiconductor quantum dot (QD) probe. The mRNA expression and protein production were quantified by counting the total pixel intensity in the region of interest (r.o.i.) surrounding single cells. As a result, the correlation was poor in a scattered plot of mRNA expression versus protein production in individual cells. These findings demonstrate that cell-to-cell differences in the translation process are also a key factor in heterogeneous gene expression. PMID- 21641987 TI - Effects of birth asphyxia on neonatal hippocampal structure and function in the spiny mouse. AB - Studies of human neonates, and in animal experiments, suggest that birth asphyxia results in functional compromise of the hippocampus, even when structural damage is not observable or resolves in early postnatal life. The aim of this study was to determine if changes in hippocampal function occur in a model of birth asphyxia in the precocial spiny mouse where it is reported there is no major lesion or infarct. Further, to assess if, as in human infants, this functional deficit has a sex-dependent component. At 37 days gestation (term=39 days) spiny mice fetuses were either delivered immediately by caesarean section (control group) or exposed to 7.5min of in utero asphyxia causing systemic acidosis and hypoxia. At 5 days of age hippocampal function was assessed ex vivo in brain slices, or brains were collected for examination of structure or protein expression. This model of birth asphyxia did not cause infarct or cystic lesion in the postnatal day 5 (P5) hippocampus, and the number of proliferating or pyknotic cells in the hippocampus was unchanged, although neuronal density in the CA1 and CA3 was increased. Protein expression of synaptophysin, brain-derived neurotrophic factor (BDNF), and the inositol trisphosphate receptor 1 (IP(3)R1) were all significantly increased after birth asphyxia, while long-term potentiation (LTP), paired pulse facilitation (PPF), and post-tetanic potentiation (PTP) were all reduced at P5 by birth asphyxia. In control P5 pups, PPF and synaptic fatigue were greater in female compared to male pups, and after birth asphyxia PPF and synaptic fatigue were reduced to a greater extent in female vs. male pups. In contrast, the asphyxia-induced increase in synaptophysin expression and neuronal density were greater in male pups. Thus, birth asphyxia in this precocial species causes functional deficits without major structural damage, and there is a sex-dependent effect on the hippocampus. This may be a clinically relevant model for assessing treatments delivered either before or after birth to protect this vulnerable region of the developing brain. PMID- 21641988 TI - Development of a human corneal epithelium model utilizing a collagen vitrigel membrane and the changes of its barrier function induced by exposing eye irritant chemicals. AB - The brief TEER (trans-epithelial electrical resistance) assay after exposing chemicals to corneal epithelium in vivo is known as a suitable method for evaluating corneal irritancy and permeability quantitatively and continuously. A collagen vitrigel membrane we previously developed is a thin (about 20 MUm thick) and transparent membrane composed of high density collagen fibrils equivalent to connective tissues in vivo, e.g. corneal Bowman's membrane. To develop such a TEER assay system in vitro utilizing a human corneal epithelial model, HCE-T cells (a human corneal epithelial cell line) were cultured on the collagen vitrigel membrane substratum prepared in a Millicell chamber suitable for TEER measurement. Human corneal epithelium model possessing 5-6 cell layers sufficient for TEER assay was successfully reconstructed on the substratum in the Millicell chamber by culturing the cells in monolayer for 2 days and subsequently in air liquid interface for 7 days. The exposure of chemicals to the model induced the time-dependent relative changes of TEER in response to the characteristic of each chemical within a few minutes. These results suggest that the TEER assay using the human corneal epithelial model is very useful for an ocular irritancy evaluation as an alternative to the Draize eye irritation test. PMID- 21641989 TI - Rapid and accurate assessment of seizure liability of drugs by using an optimal support vector machine method. AB - Drug-induced seizures are a serious adverse effect and assessment of seizure risk usually takes place at the late stage of drug discovery process, which does not allow sufficient time to reduce the risk by chemical modification. Thus early identification of chemicals with seizure liability using rapid and cheaper approaches would be preferable. In this study, an optimal support vector machine (SVM) modeling method has been employed to develop a prediction model of seizure liability of chemicals. A set of 680 compounds were used to train the SVM model. The established SVM model was then validated by an independent test set comprising 175 compounds, which gave a prediction accuracy of 86.9%. Further, the SVM-based prediction model of seizure liability was compared with various preclinical seizure assays, including in vitro rat hippocampal brain slice, in vivo zebrafish larvae assay, mouse spontaneous seizure model, and mouse EEG model. In terms of predictability, the SVM model was ranked just behind the mouse EEG model, but better than the rat brain slice and zebrafish models. Nevertheless, the SVM model has considerable advantages compared with the preclinical seizure assays in speed and cost. In summary, the SVM-based prediction model of seizure liability established here offers potential as a cheaper, rapid and accurate assessment of seizure liability of drugs, which could be used in the seizure risk assessment at the early stage of drug discovery. The prediction model is freely available online at http://www.sklb.scu.edu.cn/lab/yangsy/download/ADMET/seizure_pred.tar. PMID- 21641990 TI - From progenitors to integrated neurons: role of neurotransmitters in adult olfactory neurogenesis. AB - Adult neurogenesis is due to the persistence of pools of constitutive stem cells able to give rise to a progeny of proliferating progenitors. In rodents, adult neurogenic niches have been found in the subventricular zone (SVZ) along the lateral ventricles and in the subgranular zone of the dentate gyrus in the hippocampus. SVZ progenitors undergo a unique process of tangential migration from the lateral ventricle to the olfactory bulb (OB) where they differentiate mainly into GABAergic interneurons in the granule and glomerular layers. SVZ progenitor proliferation, migration and differentiation into fully integrated neurons, are strictly related processes regulated by complex interactions between cell intrinsic and extrinsic influences. Numerous observations demonstrate that neurotrasmitters are involved in all steps of the adult neurogenic process, but the understanding of their role is hampered by their intricate mechanism of action and by the highly complex network in which neurotransmitters work. By considering the three main steps of olfactory adult neurogenesis (proliferation, migration and integration), this review will discuss recent advances in the study of neurotransmitters, highlighting the regulatory mechanisms upstream and downstream their action. PMID- 21641991 TI - Epigallocatechin-3-gallate prevents lupus nephritis development in mice via enhancing the Nrf2 antioxidant pathway and inhibiting NLRP3 inflammasome activation. AB - Patients with lupus nephritis show an impaired oxidative status and increased levels of interleukin (IL)-1beta and IL-18, which are closely linked to inflammation and correlated with disease activity. Although epigallocatechin-3 gallate (EGCG), the major bioactive polyphenol present in green tea with antioxidant and free radical scavenging activities, has been reported to have anti-inflammatory effects by inhibiting nuclear factor-kappa B (NF-kappaB) mediated inflammatory responses in vivo, its effectiveness for the treatment of lupus nephritis is still unknown. In the present study, 12-week-old New Zealand black/white (NZB/W) F1 lupus-prone mice were treated daily with EGCG by gavage until sacrificed at 34 weeks old for clinical, pathological, and mechanistic evaluation. We found that the administration (1) prevented proteinuria, renal function impairment, and severe renal lesions; (2) increased renal nuclear factor E2-related factor 2 (Nrf2) and glutathione peroxidase activity; (3) reduced renal oxidative stress, NF-kappaB activation, and NLRP3 mRNA/protein expression and protein levels of mature caspase-1, IL-1beta, and IL-18; and (4) enhanced splenic regulatory T (Treg) cell activity. Our data clearly demonstrate that EGCG has prophylactic effects on lupus nephritis in these mice that are highly associated with its effects of enhancing the Nrf2 antioxidant signaling pathway, decreasing renal NLRP3 inflammasome activation, and increasing systemic Treg cell activity. PMID- 21641992 TI - Selective targeting of breast cancer cells through ROS-mediated mechanisms potentiates the lethality of paclitaxel by a novel diterpene, gelomulide K. AB - Defects in apoptotic pathways confer resistance to tubulin-binding agents via downregulation of caspases or overexpression of antiapoptotic factors, urging the need for novel agents acting on an alternative pathway. The purpose of this study was to investigate whether induction of ROS can induce caspase-independent cell death in breast cancer cells and thereby enhance the activity of paclitaxel. Here, we report that gelomulide K acts as a caspase-independent cell death inducing agent that synergizes with paclitaxel in breast cancer cells and has low toxicity in normal cells. Treatment with gelomulide K induced PARP-1 hyperactivation, AIF nuclear translocation, and cytoprotective autophagy. These effects were associated with increased ROS production and a decrease in cellular GSH levels in cancer cells. Furthermore, pretreatment with NAC, a precursor of intracellular GSH, effectively abrogated gelomulide K-induced caspase-independent cell death and autophagy, suggesting that ROS-mediated downstream signaling is essential to the anticancer effects of gelomulide K. Additionally, in a xenograft model, gelomulide K induced PARP-1 activation and reduced tumor growth. In terms of structure-activity relationships, analysis not only showed a correlation between ROS levels and drug activity but also highlighted the importance of the 8,14-epoxy group. Taken together, our results show that enhancement of paclitaxel activity can be achieved with gelomulide K and that the structurally relevant pharmacophore provides important insight into the development of new caspase independent cell death-inducing agents. PMID- 21641993 TI - Conserved aquaporin 4 levels associated with reduction of brain edema are mediated by estrogen in the ischemic brain after experimental stroke. AB - Aquaporin 4 (AQP4), the most abundant water channel protein in the brain, is involved in brain edema induced by ischemic insults. To evaluate whether the neuroprotective effects of estrogen are associated with AQP4 expression and edema formation, changes in AQP levels and ischemic edema were examined in the brains of male and female mice subjected to transient middle cerebral artery occlusion. Infarct volume and edema formation were markedly less in females than in males. AQP4 expression in the ischemic cortex of females was relatively well preserved, whereas it was significantly decreased in males. These effects disappeared in ovariectomized females but were reversed by estrogen replacement. Furthermore, AQP4 expression was decreased with increased brain edema in females treated with ICI182,780, an estrogen receptor antagonist. These findings suggest that the estrogen effect on the reduction of ischemic brain edema is associated with the preserved level of AQP4 that is partly mediated by estrogen receptors. PMID- 21641994 TI - Enhancing and sustaining the topical ocular delivery of fluconazole using chitosan solution and poloxamer/chitosan in situ forming gel. AB - Fungal keratitis is a serious disease that can lead to loss of vision. Unfortunately, current therapeutic options often result in poor bioavailability of antifungal agents due to protective mechanisms of the eye. The aim of this work was to evaluate the potential of a chitosan solution as well as an in situ gel-forming system comprised of poloxamer/chitosan as vehicles for enhanced corneal permeation and sustained release of fluconazole (FLU). For this, in vitro release and ex vivo corneal permeation experiments were carried out as a function of chitosan concentration from formulation containing the chitosan alone and combined with the thermosensitive polymer, poloxamer. Microdialysis was employed in a rabbit model to evaluate the in vivo performance of the formulations. The in vitro release studies showed the sustained release of FLU from the poloxamer/chitosan formulation. Ex vivo permeation studies across porcine cornea demonstrated that the formulations studied have a permeation-enhancing effect that is independent of chitosan concentration in the range from 0.5 to 1.5% w/w. The chitosan solutions alone showed the greatest ex vivo drug permeation; however, the poloxamer/chitosan formulation presented similar in vivo performance than the chitosan solution at 1.0%; both formulations showed sustained release and about 3.5-fold greater total amount of FLU permeated when compared to simple aqueous solutions of the drug. In conclusion, it was demonstrated that both the in situ gelling formulation evaluated and the chitosan solution are viable alternatives to enhance ocular bioavailability in the treatment of fungal keratitis. PMID- 21641995 TI - Caveolin-1 modulates TGF-beta1 signaling in cardiac remodeling. AB - The cardiac response to myocardial injury includes fibrotic and hypertrophic processes and a key mediator in this response is transforming growth factor-beta1 (TGF-beta1). Caveolin-1 (cav1), the main structural protein of caveolae, is an inhibitor of the TGF-beta1 signaling pathway. To examine the role of cav1 in cardiac repair, cav1 deficient (Cav1(-/-)) and wild type (WT) mice were subjected to cryoinjury of the left ventricle (LV). At baseline the two groups exhibited no inflammation, similar collagen content, and similar cardiac function. After injury, Cav1(-/-) animals displayed enhanced TGF-beta1 signaling, as reflected by a 3-fold increase in the activation of the Smad2-dependent pathway and more widespread collagen deposition in the heart. Qualitative and quantitative analyses indicated that collagen deposition peaked in the WT LV 14days after injury, accompanied by increased mRNA abundance for procol1a2 (2-fold) and procol3a1 (3-fold). Collagen deposition was further enhanced in Cav1(-/-) mice, which was accompanied by reduced expression of matrix metalloproteinases MMP-8 (3 fold) and -13 mRNA (2-fold). The levels of expression of inflammatory markers of acute phase were similar between the strains However, macrophage clearance in the damaged region was delayed in Cav1(-/-) mice. We observed a 4-fold decrease in collagen deposition in Cav1(-/-) mice injected with a cav1 scaffolding domain peptide (CSD) and a 2-fold decrease in WT mice treated with the CSD. We conclude that cav1 has a direct role in reducing TGF-beta1 signaling and as such might be an appropriate target for therapies to influence cardiac remodeling. PMID- 21641996 TI - Pyrosequencing-based expression profiling and identification of differentially regulated genes from Manduca sexta, a lepidopteran model insect. AB - Although Manduca sexta has significantly contributed to our knowledge on a variety of insect physiological processes, the lack of its genome sequence hampers the large-scale gene discovery, transcript profiling, and proteomic analysis in this biochemical model species. Here we report our implementation of the RNA-Seq cDNA sequencing approach based on massively parallel pyrosequencing, which allows us to categorize transcripts based on their relative abundances and to discover process- or tissue-specifically regulated genes simultaneously. We obtained 1,821,652 reads with an average length of 289 bp per read from fat body and hemocytes of naive and microbe-injected M. sexta larvae. After almost all (92.1%) of these reads were assembled into 19,020 contigs, we identified 528 contigs whose relative abundances increased at least 5- and 8-fold in fat body and hemocytes, respectively, after the microbial challenge. Polypeptides encoded by these contigs include pathogen recognition receptors, extracellular and intracellular signal mediators and regulators, antimicrobial peptides, and proteins with no known sequence but likely participating in defense in novel ways. We also found 250 and 161 contigs that were preferentially expressed in fat body and hemocytes, respectively. Furthermore, we integrated data from our previous study and generated a sequence database to support future gene annotation and proteomic analysis in M. sexta. In summary, we have successfully established a combined approach for gene discovery and expression profiling in organisms lacking known genome sequences. PMID- 21641997 TI - Positive virtual crossmatch with negative flow crossmatch results in two cases. AB - Pre-transplant (Tx) presence of HLA antibodies (HLA-Ab) especially donor specific antibodies (DSA) has been correlated with post-Tx rejection. While crossmatch (XM) is the specific method to identify DSA, logistical reasons prevent performing a prospective XM in all transplants. In such cases DSA as identified by solid-phase assay (SPA) are being used to perform a virtual crossmatch (VXM). We present two cases, a heart-lung transplant and a kidney transplant, for which testing detected a presumptive DSA with discordant results: a negative flow cytometric crossmatch (FXM) and a positive VXM using SPA. The subsequent investigation determined the antibody, in both cases, was presumably directed against an epitope of a HLA-B*44 antigen found on the single antigen beads (SAB) used in the SPA but not against the native form on the donor lymphocytes used in the FXM. Manufacturing of SAB beads results in denaturation of epitopes, majority of which are removed from the final product, but residual amount is present on the final product. Denaturation of majority of antigen epitopes on single antigen beads did not remove the activity of the recipient's antibodies but it did diminish the activity of positive control serum. This indicates denaturation of some of the HLA-B*44 antigen during manufacturing of the SAB may have lead to the reactivity. Antibody mediated rejection does not appear to be associated with the titer of this antibody to denatured antigen in the first case and so clinical relevance of such antibodies is unclear. Subsequently a second case of discordant FXM and VXM was identified in a potential kidney transplant patient who went on to an uneventful transplant. In this case, lymphocytes from the donor were positively shown to express HLA-B*44:02 using known anti- HLA-B*44:02 control serum. Platelets identified as HLA-B*44:02 could adsorb the anti-HLA-B*44:02 from the control serum activity but not from that of the recipient's anti- HLA-B 44 antibody adding evidence that this antibody should best be classified as a false positive finding. The presence of such an antibody if misidentified may result in unnecessary therapy being instituted or the inappropriate denial of an organ for transplantation. PMID- 21641998 TI - Non-genomic activation of adenylyl cyclase and protein kinase G by 17beta estradiol in vascular smooth muscle of the rat superior mesenteric artery. AB - The aim of the present study was to investigate the signaling mechanisms underlying the non-genomic effects of estrogen in rat superior mesenteric arteries. Isometric tension was recorded in rings with or without endothelium. Changes in cyclic nucleotide levels and protein kinase (PK) activities were measured. Localization of estrogen receptors (ER) and caveolin-1 were visualized by confocal microscopy. 17beta-Estradiol elicited a concentration-dependent relaxation. The relaxation was reduced by SQ 22536 (adenylyl cyclase inhibitor) and KT 5823 (PKG inhibitor) while ODQ (guanylyl cyclase inhibitor) and KT 5720 (PKA inhibitor) had no effect. At the physiological concentration of 1 nM, 17beta estradiol had no significant effect on relaxation but enhanced the relaxation to sodium nitroprusside. The enhancement of relaxation by 17beta-estradiol was blocked by SQ 22536 and KT 5823. Although 1 nM 17beta-estradiol or 10 nM sodium nitroprusside given alone had minimal effects on PKG activity, in their combined presence, a significant increase in PKG activity was observed. Confocal microscopy demonstrated that ERalpha and ERbeta colocalized with caveolin-1 and PKG in vascular smooth muscle cells. The present findings suggest that 17beta estradiol enhances relaxation of vascular smooth muscle of the rat superior mesenteric artery by activating adenylyl cyclase, leading to an increase in cAMP which cross activates PKG in the caveolae. No detectable increase in total cAMP level was detected as these changes occurred in the caveolae. These results are consistent with the notion that 17beta-estradiol mediates its effect in the distinct microdomains of the caveolae of the plasma membrane with colocalization of adenylyl cyclase and PKG. PMID- 21641999 TI - Production, purification and detergent exchange of isotopically labeled Bacillussubtilis cytochrome b558 (SdhC). AB - Cytochrome b558 of the gram-positive bacterium Bacillussubtilis is the membrane anchor subunit of the succinate:quinone oxidoreductase of the citric acid cycle. The cytochrome consists of the SdhC polypeptide (202 residues) and two protoheme IX groups that function in transmembrane electron transfer to menaquinone. The general structure of the cytochrome is known from extensive experimental studies and by comparison to Wolinellasuccinogenes fumarate reductase for which the X-ray crystal structure has been determined. Solution state NMR can potentially be used to identify the quinone binding site(s) and study, e.g. redox-linked, dynamics of cytochrome b558. In this work we present an efficient procedure for the isolation of preparative amounts of isotopically labeled B. subtilis cytochrome b558 produced in Escherichia coli. We have also evaluated several detergents suitable for NMR for their effectiveness in maintaining the cytochrome solubilized and intact for days at room temperature. PMID- 21642000 TI - Protective effect of propolis against oxidative stress and immunosuppression induced by oxytetracycline in rainbow trout (Oncorhynchus mykiss, W.). AB - The aim of this study was to investigate the effects of propolis on oxytetracycline (OTC)-induced oxidative stress and immunosuppression in fish. OTC (100 mg per kg-1 body weight) was orally administered to fish for 14 days. A significant elevation in the level of malondialdehyde, as an index of lipid peroxidation, and reductions in antioxidant enzyme activities (superoxide dismutase, catalase, and glutathione peroxidase) and low molecular weight antioxidant (reduced glutathione) levels were observed in the blood, liver, kidney, spleen, and heart tissues of OTC-treated fish. OTC also had a suppressive effect on specific and non-specific immune system parameters, such as leucocyte counts, oxidative radical production (nitrobluetetrazolium activity), total plasma protein and immunoglobulin levels, and phagocytic activity. Pre-treatment, post-treatment, and simultaneous treatment with propolis (50 mg per kg-1 body weight, orally) attenuated the OTC-induced oxidative stress by significantly decreasing the levels of malondialdehyde in tissues. In addition, propolis significantly increased the level of reduced glutathione and the catalase, glutathione peroxidase, and superoxide dismutase activities. Upon the administration of propolis, the suppressed immune system parameters were significantly increased in fish treated with OTC. The present results suggest that pre-treatment, post-treatment, and simultaneous administration of propolis might alleviate OTC-induced oxidative stress and immunosuppression. PMID- 21642001 TI - Viral hemorrhagic septicaemia virus (VHSV) up-regulates the cytotoxic activity and the perforin/granzyme pathway in the rainbow trout RTS11 cell line. AB - A survey of immune-relevant genes that might be up-regulated in response to viral hemorrhagic septicaemia virus (VHSV) in the rainbow trout monocyte-macrophage cell line, RTS11, unexpectedly revealed an increased expression of perforin (PRF) and granzyme (GRZ) genes, which represent components of the major cytotoxic pathway. The natural killer-enhancing factor (NKEF), also known to modulate cytotoxic activity, was up-regulated at the gene but strikingly down-regulated at protein level. The expression of these genes was not affected in head kidney leukocytes (HKLs) infected with VHSV, leading us to evaluate the potential cytotoxic activity of RTS11 and HKLs. For the first time, the cytotoxic activity of RTS11 against xenogeneic targets has been demonstrated, although this was modest relative to HKLs. Yet the activity in RTS11 was significantly increased by VHSV, as in HKLs. This cytotoxic activity elicited by viral infection appeared to require viral gene expression because inactivated VHSV failed to increase RTS11 cytotoxic activity. As for other immune functions, RTS11 cells provide a model for further studying cytotoxic activities of fish monocyte-macrophages. PMID- 21642002 TI - Transgenic expression of tilapia hepcidin 1-5 and shrimp chelonianin in zebrafish and their resistance to bacterial pathogens. AB - Recently, tilapia hepcidin (TH)1-5 was characterized, and its antimicrobial functions against several pathogens were reported. The antimicrobial functions of another shrimp antimicrobial peptide (AMP), chelonianin, were also characterized using a recombinant chelonianin protein (rcf) that was expressed by a stably transfected Chinese hamster ovary (CHO) cell line against pathogen infections in fish. The function of the overexpression of both AMPs in zebrafish muscles was not examined in previous studies. Herein, we investigated the antimicrobial functions of TH1-5 and chelonianin against Vibrio vulnificus (204) and Streptococcus agalactiae (SA48) in transgenic TH1-5 zebrafish and transgenic chelonianin zebrafish. The presence of TH1-5 and chelonianin enhanced the inhibitory ability in transgenic AMP zebrafish against the two different bacterial infections. The bacterial number of either V. vulnificus (204) or S. agalactiae (SA48) had decreased at 96 h after injection into transgenic AMP zebrafish muscle compared to non-transgenic zebrafish muscle. Additionally, immune-related gene expressions analyzed by real-time PCR studies showed the modulation of several genes including interleukin (IL)-10, IL-22, IL-26, MyD88, Toll-like receptor (TLR)-1, TLR-3, TLR-4, nuclear factor (NF)-kappaB, tumor necrosis factor (TNF)-alpha, and lysozyme, and significant differences were found between transgenic AMP zebrafish and wild-type zebrafish injected with PBS at 1 24 h. These results suggest that several immune-related gene expressions were induced in transgenic TH1-5 and chelonianin zebrafish which effectively inhibited bacterial growth. The survival rate dropped to 86.6% in transgenic chelonianin zebrafish after 28 days of infection compared of the 50% survival rate in transgenic TH1-5 zebrafish after 28 days of infection. Overall, these results indicate that TH1-5 and chelonianin possess the potential to be novel candidate genes for aquaculture applications to treat fish diseases. PMID- 21642003 TI - Molecular characterization, expression and functional analysis of a nuclear oligomerization domain proteins subfamily C (NLRC) in Japanese flounder (Paralichthys olivaceus). AB - Pattern recognition receptors (PRRs) are involved in the effective innate defense against several microbes. Here, we identified a nucleotide-oligomerization domain (NOD)-like receptor subfamily C (NLRC) from Japanese flounder (Paralichthys olivaceus). Full-length transcript of JfNLRC is composed of 3976 bp encoding a protein of 1175 deduced amino acid residues. The presence of a signature nucleotide-binding domain (NACHT) and leucine-rich repeated domain (LRR) suggested that the protein is a member of the NLR family. Interestingly, its C terminus presents an extra PRY/SPRY (B30.2) domain similar to fish in the Trim (finTrim) family. A phylogenic tree of JfNLRC revealed that full-length JfNLRC diverged from the NOD1 and NOD2 clusters, and the NACHT domain in JfNLRC was clustered within the NLRC3 group. Stimulation by formalin-killed Edwardsiella tarda, Streptococcus iniae, and lipopolysaccharide (LPS) showed that the JfNLRC expression was raised a few hours after stimulation, suggesting this novel protein is involved in the immediate response against both Gram-positive and Gram negative bacteria. Furthermore, the IL-1beta mRNA expression level in JfNLRC-over expressing HINAE cells was significantly increased, when compared to a control, after LPS-stimulation and E. tarda infection. These results suggested that JfNLRC probably induced IL-1beta gene expression mediated by LPS-stimulation. PMID- 21642004 TI - DISC1 is associated with cortical thickness and neural efficiency. AB - BACKGROUND: Disrupted in schizophrenia 1 (DISC1) is known to play a major role during brain development and is a candidate gene for schizophrenia. Cortical thickness is highly heritable and several MRI studies have shown widespread reductions of cortical thickness in patients with schizophrenia. Here, we investigated the effects of variation in DISC1 on cortical thickness. In a subsequent analysis we tested whether the identified DISC1 risk variant is also associated with neural activity during working memory functioning. METHODS: We acquired structural MRI (sMRI), functional MRI (fMRI) and genotype data from 96 healthy volunteers. Separate cortical statistical maps for five single nucleotide polymorphisms (SNP) of DISC1 were generated to detect differences of cortical thickness in genotype groups across the entire cortical surface. Working-memory related load-dependent activation was measured during the Sternberg Item Recognition Paradigm and analyzed using a region-of-interest approach. RESULTS: Phe allele carriers of the DISC1 SNP Leu607Phe had significantly reduced cortical thickness in the left supramarginal gyrus compared to Leu/Leu homozygotes. Neural activity in the left dorsolateral prefrontal cortex (DLPFC) during working memory task was increased in Phe allele carriers, whereas working memory performance did not differ between genotype groups. CONCLUSIONS: This study provides convergent evidence for the effect of DISC1 risk variants on two independent brain-based intermediate phenotypes of schizophrenia. The same risk variant was associated with cortical thickness reductions and signs of neural inefficiency during a working memory task. Our findings provide further evidence for a neurodevelopmental model of schizophrenia. PMID- 21642005 TI - Clustering method for estimating principal diffusion directions. AB - Diffusion tensor magnetic resonance imaging (DTMRI) is a non-invasive tool for the investigation of white matter structure within the brain. However, the traditional tensor model is unable to characterize anisotropies of orders higher than two in heterogeneous areas containing more than one fiber population. To resolve this issue, high angular resolution diffusion imaging (HARDI) with a large number of diffusion encoding gradients is used along with reconstruction methods such as Q-ball. Using HARDI data, the fiber orientation distribution function (ODF) on the unit sphere is calculated and used to extract the principal diffusion directions (PDDs). Fast and accurate estimation of PDDs is a prerequisite for tracking algorithms that deal with fiber crossings. In this paper, the PDDs are defined as the directions around which the ODF data is concentrated. Estimates of the PDDs based on this definition are less sensitive to noise in comparison with the previous approaches. A clustering approach to estimate the PDDs is proposed which is an extension of fuzzy c-means clustering developed for orientation of points on a sphere. MDL (Minimum description length) principle is proposed to estimate the number of PDDs. Using both simulated and real diffusion data, the proposed method has been evaluated and compared with some previous protocols. Experimental results show that the proposed clustering algorithm is more accurate, more resistant to noise, and faster than some of techniques currently being utilized. PMID- 21642006 TI - Confidence intervals for the substitution number in the nucleotide substitution models. AB - In the nucleotide substitution model for molecular evolution, a major task in the exploration of an evolutionary process is to estimate the substitution number per site of a protein or DNA sequence. The usual estimators are based on the observation of the difference proportion of the two nucleotide sequences. However, a more objective approach is to report a confidence interval with precision rather than only providing point estimators. The conventional confidence intervals used in the literature for the substitution number are constructed by the normal approximation. The performance and construction of confidence intervals for evolutionary models have not been much investigated in the literature. In this article, the performance of these conventional confidence intervals for one-parameter and two-parameter models are explored. Results show that the coverage probabilities of these intervals are unsatisfactory when the true substitution number is small. Since the substitution number may be small in many situations for an evolutionary process, the conventional confidence interval cannot provide accurate information for these cases. Improved confidence intervals for the one-parameter model with desirable coverage probability are proposed in this article. A numerical calculation shows the substantial improvement of the new confidence intervals over the conventional confidence intervals. PMID- 21642007 TI - Nitration of tryptophan in ribosomal proteins is a novel post-translational modification of differentiated and naive PC12 cells. AB - Neuron growth factor (NGF) signaling in PC12 cell, which is derived from pheochromocytoma of rat adrenal medulla, induces expression of neuronal nitric oxide synthase (nNOS) and nitric oxide (NO) production. Subsequently, NO causes differentiation of PC12 cell to neuronal cell with morphological changes, such as neurite extension. In this study, we showed that 6-nitrotryptophan-containing proteins were produced in PC12 cell (naive PC12 cell) and/or NGF-induced PC12 cell (differentiated PC12 cell). Western blot analysis of the protein extract of naive PC12 cell and differentiated PC12 cell using anti 6-nitrotryptophan antibody showed several immunoreactive bands, which were subsequently subjected to trypsin digestion and LC-ESI-MS-MS analysis. The peptides from five ribosomal proteins, namely, 60S ribosomal protein L7 (Trp154), 60S acidic ribosomal protein P1 (Trp43), 40S ribosomal protein S2 (Trp60), 40S ribosomal protein S6 (Trp45), and 40S ribosomal protein S19 (Trp52), were identified as nitrotryptophan residue containing proteins with significant ion score levels (p<0.05). Among these, tryptophan nitration was observed only in differentiated PC12 cell for S19 protein, and only in naive PC12 cell for L7 protein. Tryptophan nitration of the other ribosomal proteins P1, S2, and S6 was observed in both naive and differentiated PC12 cells. The positive signal of nitrotryptophan-containing proteins in the Western blotting around 16 kDa (Band 1), which includes 40S ribosomal protein S19, was suppressed by treatment with NOS inhibitor, L-NAME. The tryptophan nitration of 40S ribosomal protein was not observed by LC-ESI-MS MS analysis of this sample. This is the first study to identify several specific sites of nitrated tryptophan on proteins not only in viable culture cells but also in a physiological process: cell differentiation. PMID- 21642008 TI - Cell surface thiol isomerases may explain the platelet-selective action of S nitrosoglutathione. AB - S-nitrosoglutathione (GSNO) at low concentration inhibits platelet aggregation without causing vasodilation, suggesting platelet-selective nitric oxide delivery. The mechanism of this selectivity is unknown, but may involve cell surface thiol isomerases, in particular protein disulphide isomerase (csPDI) (EC 5.3.4.1). We have now compared csPDI expression and activity on platelets, endothelial cells and vascular smooth muscle cells, and the dependence on thiol reductase activity of these cell types for NO uptake from GSNO. csPDI expression was measured by flow cytometry and its reductase activity using the pseudosubstrate dieosin glutathione disulphide. This activity assay was adapted and validated for 96-well plate format. Flow cytometry revealed csPDI on all three cell types, but percentage positivity of expression was higher on platelets than on vascular cells. Consistent with this, thiol isomerase-related reductase activity was higher on platelets (P<0.01), and cellular activation (with either phorbol myristate acetate or ionomycin) increased csPDI activity on both platelets and smooth muscle cells, but not on endothelium. Intracellular NO delivery from GSNO was greater in platelets than in vascular cells (P<0.002), and was more sensitive to thiol isomerase inhibition using phenylarsine oxide (P<0.05). Increased surface thiol isomerase activity on platelets, compared with cells of the vascular wall, may explain the platelet-selective actions of GSNO and help define its antithrombotic potential. PMID- 21642009 TI - Nitric oxide regulates pulmonary vascular smooth muscle cell expression of the inducible cAMP early repressor gene. AB - Nitric oxide (NO) regulates vascular smooth muscle cell (VSMC) structure and function, in part by activating soluble guanylate cyclase (sGC) to synthesize cGMP. The objective of this study was to further characterize the signaling mechanisms by which NO regulates VSMC gene expression using transcription profiling. DNA microarrays were hybridized with RNA extracted from rat pulmonary artery smooth muscle cells (RPaSMC) exposed to the NO donor compound, S-nitroso glutathione (GSNO). Many of the genes, whose expression was induced by GSNO, contain a cAMP-response element (CRE), of which one encoded the inducible cAMP early repressor (ICER). sGC and cAMP-dependent protein kinase, but not cGMP dependent protein kinase, were required for NO-mediated phosphorylation of CRE binding protein (CREB) and induction of ICER gene expression. Expression of a dominant-negative CREB in RPaSMC prevented the NO-mediated induction of CRE dependent gene transcription and ICER gene expression. Pre-treatment of RPaSMC with the intracellular calcium (Ca(2+)) chelator, BAPTA-AM, blocked the induction of ICER gene expression by GSNO. The store-operated Ca(2+) channel inhibitors, 2 ABP, and SKF-96365, reduced the GSNO-mediated increase in ICER mRNA levels, while 2-ABP did not inhibit GSNO-induced CREB phosphorylation. Our results suggest that induction of ICER gene expression by NO requires both CREB phosphorylation and Ca(2+) signaling. Transcription profiling of RPaSMC exposed to GSNO revealed important roles for sGC, PKA, CREB, and Ca(2+) in the regulation of gene expression by NO. The induction of ICER in GSNO-treated RPaSMC highlights a novel cross-talk mechanism between cGMP and cAMP signaling pathways. PMID- 21642010 TI - Kinetic modelling reveals current limitations in the production of ethanol from xylose by recombinant Saccharomyces cerevisiae. AB - Saccharomyces cerevisiae lacks the ability to ferment the pentose sugar xylose that is the second most abundant sugar in nature. Therefore two different xylose catabolic pathways have been heterologously expressed in S. cerevisiae. Whereas the xylose reductase (XR)-xylitol dehydrogenase (XDH) pathway leads to the production of the by-product xylitol, the xylose isomerase (XI) pathway results in significantly lower xylose consumption. In this study, kinetic models including the reactions ranging from xylose transport into the cell to the phosphorylation of xylulose to xylulose 5-P were constructed. They were used as prediction tools for the identification of putative targets for the improvement of xylose utilization in S. cerevisiae strains engineered for higher level of the non-oxidative pentose phosphate pathway (PPP) enzymes, higher xylulokinase and inactivated GRE3 gene encoding an endogenous NADPH-dependent aldose reductase. For both pathways, the in silico analyses identified a need for even higher xylulokinase (XK) activity. In a XR-XDH strain expressing an integrated copy of the Escherichia coli XK encoding gene xylB about a six-fold reduction of xylitol formation was confirmed under anaerobic conditions. Similarly overexpression of the xylB gene in a XI strain increased the aerobic growth rate on xylose by 21%. In contrast to the in silico predictions, the aerobic growth also increased 24% when the xylose transporter gene GXF1 from Candida intermedia was overexpressed together with xylB in the XI strain. Under anaerobic conditions, the XI strains overexpressing xylB gene and the combination of xylB and GFX1 genes consumed 27% and 37% more xylose than the control strain. PMID- 21642011 TI - Structure-activity relationship of ibogaine analogs interacting with nicotinic acetylcholine receptors in different conformational states. AB - The interaction of ibogaine analogs with nicotinic acetylcholine receptors (AChRs) in different conformational states was studied by functional and structural approaches. The results established that ibogaine analogs: (a) inhibit (+/-)-epibatidine-induced Ca2+ influx in human embryonic muscle AChRs with the following potency sequence (IC(50) in MUM): (+/-)-18-methylaminocoronaridine (5.9+/-0.3)~(+/-)-18-methoxycoronaridine (18-MC) (6.8+/-0.8)>(-)-ibogaine (17+/ 3)~(+)-catharanthine (20+/-1)>(+/-)-albifloranine (46+/-13), (b) bind to the [3H]TCP binding site with higher affinity when the Torpedo AChR is in the desensitized state compared to that in the resting state. Similar results were obtained using [3H]18-MC. These and docking results suggest a steric interaction between TCP and ibogaine analogs for the same site, (c) enhance [3H]cytisine binding to resting but not to desensitized AChRs, with desensitizing potencies (apparent EC50) that correlate very well with the pK(i) values in the desensitized state, and (d) there are good bilinear correlations between the ligand molecular volumes and their affinities in the desensitized and resting states, with an optimal volume of ~345 A3 for the ibogaine site. These results indicate that the size of the binding sites for ibogaine analogs, located between the serine and nonpolar rings and shared with TCP, is an important structural feature for binding and for inducing desensitization. PMID- 21642012 TI - Development and maturation in the nereidid polychaetes Platynereis dumerilii and Nereis succinea exposed to xenoestrogens. AB - Few studies link biochemical, cellular and whole animal effects of toxic compounds with growth and reproductive output on invertebrate model organisms. Thus, we explore the effects of xenoestrogens on nereid worms. Larvae of Platynereis dumerilii exposed to estradiol (E(2)) ethynylestradiol (EE(2)) and nonyplhenol (NP) observing the effects on growth, primordial germ cell (PGC) proliferation and maturation. In addition, a single exposure study was performed with a 50 days latency period on adult worms of Nereis succinea. Since reduced glutathione (GSH) is required in detoxification of NP and is the precursor of the spawning pheromone nereithione (CSSG) in N. succinea, we analysed how the estrogenic chemical NP affects GSH concentrations. PGC were not affected by exposure to E(2) and EE(2) from 24hpf to 6 days. Chronic exposure of P. dumerilii with NP over the full life cycle did not influence segment proliferation. Mature females that developed, even at high concentrations, were able to spawn and successful fertilization occurred. However, at high NP levels no P. dumerilii males matured. A significant decline of GSH can be seen in N. succinea males upon treatment with NP, but not in females, indicating that females stabilize GSH levels even in stress situations. This study shows some results that link the foundation to causally integrate toxic exposure to xenoestrogens with development, growth and reproductive outputs in nereidid polychaetes. PMID- 21642013 TI - Combining PubMed knowledge and EHR data to develop a weighted bayesian network for pancreatic cancer prediction. AB - In this paper, we propose a novel method that combines PubMed knowledge and Electronic Health Records to develop a weighted Bayesian Network Inference (BNI) model for pancreatic cancer prediction. We selected 20 common risk factors associated with pancreatic cancer and used PubMed knowledge to weigh the risk factors. A keyword-based algorithm was developed to extract and classify PubMed abstracts into three categories that represented positive, negative, or neutral associations between each risk factor and pancreatic cancer. Then we designed a weighted BNI model by adding the normalized weights into a conventional BNI model. We used this model to extract the EHR values for patients with or without pancreatic cancer, which then enabled us to calculate the prior probabilities for the 20 risk factors in the BNI. The software iDiagnosis was designed to use this weighted BNI model for predicting pancreatic cancer. In an evaluation using a case-control dataset, the weighted BNI model significantly outperformed the conventional BNI and two other classifiers (k-Nearest Neighbor and Support Vector Machine). We conclude that the weighted BNI using PubMed knowledge and EHR data shows remarkable accuracy improvement over existing representative methods for pancreatic cancer prediction. PMID- 21642014 TI - Certolizumab pegol for active Crohn's disease: a placebo-controlled, randomized trial. AB - BACKGROUND & AIMS: Certolizumab pegol (CZP) is a pegylated-conjugated Fab' against tumor necrosis factor (TNF). Additional data are needed regarding the efficacy of induction therapy with CZP in active Crohn's disease (CD). METHODS: A placebo-controlled trial evaluated the efficacy of CZP therapy in 439 adults with moderate to severe CD naive to anti-TNF therapy. Patients were randomized to receive CZP (400 mg subcutaneously) or placebo at weeks 0, 2, and 4. The primary end point was clinical remission at week 6. RESULTS: Clinical remission rates at week 6 in the CZP and placebo groups were 32% and 25% (P = .174), respectively. Remission rates at weeks 2 and 4 in the CZP and placebo groups were 23% and 16% (P = .033) and 27% and 19% (P = .063), respectively. Clinical response rates at weeks 2, 4, and 6 in the CZP and placebo groups were 33% and 20% (P = .001), 35% and 26% (P = .024), and 41% and 34% (P = .179), respectively. There were significantly greater rates of clinical remission at week 6 for CZP in patients with increased concentrations of C-reactive protein (>=5 mg/L) at entry. Serious adverse events developed in 5% and 4% of patients in the CZP and placebo groups, respectively. CONCLUSIONS: The primary end point did not reach statistical significance. Significant differences between CZP and placebo were observed in patients who had increased concentrations of C-reactive protein when the study began. Future clinical trials should emphasize the treatment of patients who have objective evidence of inflammation in addition to symptoms of active disease. PMID- 21642015 TI - Ratio of visceral to subcutaneous fat area is a biomarker of complicated Crohn's disease. AB - BACKGROUND & AIMS: Fat wrapping and mesenteric hypertrophy are characteristics of Crohn's disease (CD). In patients with CD, mesenteric adipose tissue releases higher levels of adiponectin, which could up-regulate production of tumor necrosis factor-alpha and increase the risk for aggressive disease. We investigated whether a higher ratio of visceral to subcutaneous fat was associated with complicated (fistulating or stricturing) CD. METHODS: We identified patients with a confirmed diagnosis of CD who had computed tomography scans of their abdomens (n = 50). Areas of subcutaneous and visceral fat were measured in 1 cross-sectional scan that was taken at the level of the umbilicus. The mesenteric fat index (MFI), defined as the ratio of areas of visceral to subcutaneous fat, was compared between patients with complicated (strictures and fistulas) and inflammatory CD. RESULTS: The mean age of the patients with complications (n = 29) was 49.3 +/- 15.6 years, and in patients with inflammatory CD (n = 21) it was 37.7 +/- 19.1 years. The MFI was significantly higher (P = .001) in patients with complicated disease (0.67 +/- 0.29) than in those with uncomplicated disease (0.23 +/- 0.10) and was the only variable that remained significantly different on multivariate analysis. The area under the receiver operating curve for the MFI was 0.95 (95% confidence interval, 0.89-1.0), and an MFI of 0.29 identified patients with complicated CD with 93% sensitivity and 81% specificity. CONCLUSIONS: A high ratio of areas of visceral to subcutaneous fat (MFI) is a marker of aggressive CD. Further studies are needed to determine the roles of adipose tissue in pathogenesis of CD. PMID- 21642016 TI - To the editor-biventricular pacing in hypertrophic obstructive cardiomyopathy. PMID- 21642017 TI - Proanthocyanidins from grape seeds modulates the nuclear factor-kappa B signal transduction pathways in rats with TNBS-induced recurrent ulcerative colitis. AB - The aim of this study was to elucidate the molecular mechanisms involved in the therapeutic effects of proanthocyanidins from grape seeds (GSPE) on recurrent ulcerative colitis (UC) in rats. GSPE in doses of 100, 200, and 400mg/kg were intragastrically administered per day for 7 days after recurrent colitis was twice-induced by TNBS. The levels of GSH, as well as the activity of GSH-Px and SOD in colon tissues were measured by biochemical methods. The expression levels of tumor necrosis factor-alpha (TNF-alpha) and the nuclear translocation levels of nuclear factor-kappa B (NF-kappaB) in the colon tissues were measured by enzyme-linked immunosorbent assay methods. Western blotting analysis was used to determine the protein expression levels of inhibitory kappa B-alpha (IkappaBalpha), inhibitor kappa B kinase (IKKalpha/beta), phosphorylated IkappaBalpha and phosphorylated IKKalpha/beta. GSPE treatment was associated with a remarkable increased the activity of GSH-Px and SOD with GSH levels in TNBS induced recurrent colitis rats as compared to the model group. GSPE also significantly reduced the expression levels of TNF-alpha, p-IKKalpha/beta, p IkappaBalpha and the translocation of NF-kappaB in the colon mucosa. GSPE exerted a protective effect on recurrent colitis in rats by modifying the inflammatory response and promoting damaged tissue repair to improve colonic oxidative stress. Moreover, GSPE inhibited the TNBS-induced inflammatory of recurrent colitis though blocking NF-kappaB signaling pathways. PMID- 21642018 TI - Pharmacokinetic/pharmacodynamic analyses of chymase inhibitor SUN13834 in NC/Nga mice and prediction of effective dosage for atopic dermatitis patients. AB - A chymase inhibitor SUN13834 has been shown to improve skin condition in animal models for atopic dermatitis. In the present study, effective dosages of SUN13834 for atopic dermatitis patients were predicted by pharmacokinetic/pharmacodynamic (PK/PD) analyses of SUN13834 in NC/Nga mice, which spontaneously develop atopic dermatitis-like skin lesions. For the PK/PD analyses, we utilized the minimum effective plasma concentration of unbound SUN13834 in late-phase reaction of trinitrochlorobenzene (TNCB)-induced biphasic dermatitis in mice, based on the assumption that the minimum effective plasma concentrations are the same among the two animal models. In late-phase reaction of biphasic dermatitis, SUN13834 was most effective when its plasma concentration was highest at the elicitation, and the minimum effective plasma concentration of unbound SUN13834 at the elicitation was calculated to be 0.13-0.2 ng/mL. Oral administration of SUN13834 improved dermatitis in NC/Nga mice at 15 mg/kg (twice a day; bid) and 30 mg/kg (once a day; qd), but not at 60 mg/kg (every other day; eod). At the three dosages, the duration times over the plasma level of 0.13-0.2 ng/mL were 16.1 20.3, 10.7-12.2 and 7.8-8.8h, respectively, suggesting an importance of maintenance of the minimum effective plasma concentration for at least about 10 12h. The clinical effective dosage predicted in this paper is also discussed in relation to a recently conducted Phase 2a study. PMID- 21642019 TI - Inflammation and cardiorespiratory control: the role of the vagus nerve. AB - Inflammation and immunity have been implicated in a wide variety of diseases and disorders ranging from asthma to cardiovascular disease to hemorrhagic shock. In this review we will briefly consider the evidence for the neural concomitants of immunomodulation. First, we will briefly review the anatomy and physiology of the cardiorespiratory system. Then we will review the anatomy and physiology of neural-immune communication. The nucleus of the solitary tract is a site of integration of both the afferent and efferent neural regulation of the cardiorespiratory as well as the immune system. Then we will provide an overview of what is known about neuroimmunomodulation from both animal and human studies including neuroimaging and clinical studies. Finally, we will discuss a possible role of this neural circuitry in asthma related health disparities. PMID- 21642020 TI - Prenatal development of the pulmonary surfactant system and the influence of hypoxia. AB - Pulmonary surfactant fulfils diverse functions at the lung air-liquid interface of all air-breathing vertebrates. Neurohormonal regulation of surfactant synthesis and secretion is highly conserved among non-mammalian amniotes. Although the pattern of surfactant lipid maturation is similar among species, the onset and completion differ dramatically. These differences are apparently not determined by phylogeny, but may relate to the timing of development of relative hypoxia as an embryo develops, which is related to birthing strategy. We have proposed that hypoxia is an evolutionary drive for differential surfactant development among species. In mammalian and non-mammalian models, hypoxia induces fetal growth restriction. Depending on the timing of the insult, this may be associated with an acceleration or deceleration of surfactant development. The hypoxic effect may be mediated via hormonal and growth factors, such as glucocorticoids and VEGF. However, the multifactorial nature of mammalian growth restriction models complicates the mechanistic interpretations. Hence, less complex oviparous animal models are required, in which hypoxia can be isolated from maternal influences. PMID- 21642021 TI - Uniform cell colonization of porous 3-D scaffolds achieved using radial control of surface chemistry. AB - Uniform cellular distribution is a prerequisite to forming tissue within porous scaffolds, but the seeding process often results in preferential adhesion of cells at the periphery. We develop a vapour phase coating strategy which is readily applicable to any porous solid to provide a uniform cellular distribution. Plasma polymerized allyl amine (ppAAm) is used to form a thin nitrogen-containing coating throughout porous three-dimensional (3-D) poly(d,l lactic acid) scaffolds. Subsequent controlled deposition of a hydrocarbon plasma polymerized hexane (ppHex) allows control of the fibroblast penetration into these porous 3-D objects. In order to optimize the coating conditions, a planar pinhole model of plasma penetration into pores is developed to rapidly measure deposit penetration using picolitre water contact angle measurement. Sufficiently good control over the plasma deposition within the porous scaffold is achieved using this approach to superimpose a relatively cell-repellent ppHex coating at the scaffold periphery onto the ppAAm-coated core, with a chemical gradient between the two. This 3-D chemical gradient encourages 3T3 fibroblast cells to adhere homogeneously from the periphery to the centre, when balanced by the tortuousity of the pore structure, which cells experience when passing from the surrounding medium to the centre. PMID- 21642022 TI - The realistic prediction of oxygen transport in a tissue-engineered scaffold by introducing time-varying effective diffusion coefficients. AB - An adequate oxygen supply is one of the most important factors needed in order to regenerate or engineer thick tissues or complex organs. To devise a method for maximizing the amount of oxygen available to cells, it is necessary to understand and to realistically predict oxygen transport within an engineered tissue. In this study, we focused on the fact that oxygen transport through a tissue engineered scaffold may vary with time as cells proliferate. To confirm this viewpoint, effective oxygen diffusion coefficients (D(e)(,)(s)) of scaffolds were deduced from experimental measurements and simulations of oxygen-concentration profiles were performed using these D(e)(,)(s) values in a two-dimensional (2-D) perfusion model. The results of this study indicate that higher porosity, hydraulic permeability and interconnectivity of scaffolds with no cells are responsible for the prominent diffusion capability quantified using D(e)(,)(s). On the other hand, the D(e)(,)(s) of scaffolds with cells has a negative linear relationship with cell density. Cell proliferation with time leads to a significant decrease in oxygen concentration in the 2-D perfusion model. This result demonstrates the gradual restriction of oxygen transport in a porous scaffold during cell culture. Therefore, the realistic prediction of oxygen transport using a time-varying D(e)(,)(s) will provide an appropriate basis for designing optimal transport networks within a thick scaffold. PMID- 21642023 TI - VEGF-A and FGF gene therapy accelerate healing of ischemic colonic anastomoses (experimental study). AB - BACKGROUND: Reducing ischemic damage is one of the goals of surgery. The aim of this study was to apply human VEGF-A and FGF-2 DNA-mediated gene therapy in order to identify their effects in the healing of ischemic colon anastomoses and eliminating the negative effects of ischemia. METHODS: Forty male Wistar albino rats weighing 250-280 g were divided into five equal groups (n = 8) as follows: group 1: control, ischemic left colonic anastomosis; group; 2: ischemic left colonic anastomosis with control plasmid delivery; group 3: ischemic left colonic anastomosis with VEGF plasmid delivery; group 4: ischemic left colonic anastomosis with FGF plasmid delivery; group 5: ischemic left colonic anastomosis with VEGF and FGF plasmid delivery. All rats were sacrificed on the 4th postoperative day. Anastomosis burst pressures were measured for mechanical examination of anastomosis. Tissue hydroxyprolin, VEGF and FGF levels were determined as biochemical parameters. Necrosis, epithelisation, inflammatory processes, fibroblastic activity, collagen deposition and neovascularisation at the anastomic site were studied. RESULTS: VEGF, FGF and combined therapy significantly accelerated many of the histological parameters of healing, including fibroblast activation, collagen deposition, and angiogenesis, and augmented the levels of hydroxyproline and bursting pressure. CONCLUSIONS: This is the first study to use gene therapy with growth factors for the healing of ischemic colonic anastomosis. This therapy can be effectively used in increasing ischemic anastomosis wound healing. PMID- 21642024 TI - Co-cultivation of mutant Penicillium oxalicum SAU(E)-3.510 and Pleurotus ostreatus for simultaneous biosynthesis of xylanase and laccase under solid-state fermentation. AB - Co-cultivation of mutant Penicillium oxalicum SAU(E)-3.510 and Pleurotus ostreatus MTCC 1804 was evaluated for the production of xylanase-laccase mixture under solid-state fermentation (SSF) condition. Growth compatibility between mutant P. oxalicum SAU(E)-3.510 and white rot fungi (P. ostreatus MTCC 1804, Trametes hirsuta MTCC 136 and Pycnoporus sp. MTCC 137) was analyzed by growing them on potato dextrose agar plate. Extracellular enzyme activities were determined spectrophotometrically. Under derived conditions, paired culturing of mutant P. oxalicum SAU(E)-3.510 and P. ostreatus MTCC 1804 resulted in 58% and 33% higher levels of xylanase and laccase production, respectively. A combination of sugarcane bagasse and black gram husk in a ratio of 3:1 was found to be the most ideal solid substrate and support for fungal colonization and enzyme production during co-cultivation. Maximum levels of xylanase (8205.31 +/- 168.31 IU g(-1)) and laccase (375.53 +/- 34.17 IU g(-1)) during SSF were obtained by using 4 g of solid support with 80% of moisture content. Furthermore, expressions of both xylanase and laccase were characterized during mixed culture by zymogram analysis. Improved levels of xylanase and laccase biosynthesis were achieved by co-culturing the mutant P. oxalicum SAU(E)-3.510 and P. ostreatus MTCC 1804. This may be because of efficient substrate utilization as compared to their respective monocultures in the presence of lignin degradation compounds because of synergistic action of xylanase and laccase. Understanding and developing the process of co-cultivation appears productive for the development of mixed enzyme preparation with tremendous potential for biobleaching. PMID- 21642025 TI - Differential expression of Trypanosoma cruzi I associated with clinical forms of Chagas disease: overexpression of oxidative stress proteins in acute patient isolate. AB - Chagas disease has a variable clinical course with different manifestations and heterogenous geographical distribution. Some studies suggest that this clinical variability could be influenced by the genetic variability of T. cruzi. Here we present the differential protein expression among trypomastigotes and amastigotes of T. cruzi group I isolates from patients with acute and chronic form of Chagas disease from Santander, Colombia. A total of 29 proteins were identified by MALDI TOF and LC-MS/MS; twenty in trypomastigote and nine in amastigote stage. The 29 proteins identified were grouped in 7 functional categories: 1) metabolism 31%, 2) assembly of cytoskeleton 13.7%, 3) protein destination 13.7%, 4) defenses antioxidants 20.6%, 5) protein synthesis and cellular cycle 13.7%, 6) catabolism 6.8%, and 7) adhesion 3.4%. Tryparedoxin peroxidase, lipoamide dehydrogenase, tyrosine amino transferase and HSP70 were overexpressed in the acute Chagas isolate. Tryparedoxin peroxidase overexpression in the acute isolate was confirmed by Western blot analysis. Most of these proteins are associated with resistance to oxidative stress facilitating their survival within host cells. Therefore, these proteins may represent virulence factors associated with the development of the acute form of the disease and could be used as biomarkers of the clinical course of disease and as drug targets. PMID- 21642026 TI - Renal fibrosis and proteomics: current knowledge and still key open questions for proteomic investigation. AB - Renal tubulo-interstitial fibrosis is a non-specific process, representing the final common pathway for all kidney diseases, irrespective of their initial cause, histological injury, or etiology, leading to gradual expansion of the fibrotic mass which destroys the normal structure of the tissue and results in organ dysfunction and, ultimately, in end-stage organ failure. Proteomic studies of the fibrotic pathophysiological mechanisms have been performed in cell cultures, animal models and human tissues, addressing some of the key issues. This article will review proteomic contribution to the raising current knowledge on renal fibrosis biology and also mention seminal open questions to which proteomic techniques and proteomists could fruitfully contribute. PMID- 21642027 TI - Tibial compression is anabolic in the adult mouse skeleton despite reduced responsiveness with aging. AB - The ability of the skeleton to adapt to mechanical stimuli diminishes with age in diaphyseal cortical bone, making bone formation difficult for adults. However, the effect of aging on adaptation in cancellous bone, tissue which is preferentially lost with age, is not well characterized. To develop a model for early post-menopausal women and determine the effect of aging on cancellous bone adaptation in the adult mouse skeleton, in vivo tibial compression was applied to adult (26 week old) osteopenic female mice using loading parameters, peak applied load and peak diaphyseal strain magnitude, that were previously found to be osteogenic in young, growing (10 week old) mice. A Load-Matched group received the same peak applied loads (corresponding to +2100 MUepsilon at the medial diaphysis of the tibia) and a Strain-Matched group received the same peak diaphyseal strains (+1200 MUepsilon, requiring half the load) as the young mice. The effects of mechanical loading on bone mass and architecture in adult mice were assessed using micro-computed tomography and in vivo structural stiffness measures. Adaptation occurred only in the Load-Matched group in both the metaphyseal and diaphyseal compartments. Cancellous bone mass increased 54% through trabecular thickening, and cortical area increased 41% through medullary contraction and periosteal expansion. Adult mice were able to respond to an anabolic stimulus and recover bone mass to levels seen in growing mice; however, the adaptive response was reduced relative to that in 10 week old female mice for the same applied load. Using this osteogenic loading protocol, other factors affecting pathological bone loss can be addressed using an adult osteopenic mouse model. PMID- 21642028 TI - 20 years of the bioengineering Department at the Politecnico, Milano, and beyond... AB - To celebrate the 20th anniversary of the first Bioengineering Department in Italy at the Politecnico di Milano, a number of articles in this issue highlight the department's achievements. The department was eventually founded in 1990 after many years of research and didactic activity in the area that officially began in 1969 when the first teaching course, called biological electronics, was offered in the electronic engineering specialization. PMID- 21642029 TI - Photonics for life. AB - Light is strictly connected with life, and its presence is fundamental for any living environment. Thus, many biological mechanisms are related to light interaction or can be evaluated through processes involving energy exchange with photons. Optics has always been a precious tool to evaluate molecular and cellular mechanisms, but the discovery of lasers opened new pathways of interactions of light with biological matter, pushing an impressive development for both therapeutic and diagnostic applications in biomedicine. The use of light in different fields has become so widespread that the word photonics has been utilized to identify all the applications related to processes where the light is involved. The photonics area covers a wide range of wavelengths spanning from soft X-rays to mid-infrared and includes all devices related to photons as light sources, optical fibers and light guides, detectors, and all the related electronic equipment. The recent use of photons in the field of telecommunications has pushed the technology toward low-cost, compact, and efficient devices, making them available for many other applications, including those related to biology and medicine where these requirements are of particular relevance. Moreover, basic sciences such as physics, chemistry, mathematics, and electronics have recognized the interdisciplinary need of biomedical science and are translating the most advanced researches into these fields. The Politecnico school has pioneered many of them,and this article reviews the state of the art of biomedical research at the Politecnico in the field internationally known as biophotonics. PMID- 21642030 TI - Functional evaluation and rehabilitation engineering. AB - Life is complex and all about movement, which allows us to interact with the environment and communicate with each other. The human nervous system is capable of performing a simultaneous and integrated control of 100-150 mechanical degrees of freedom of movement in the body via tensions generated by about 700 muscles. In its widest context, movement is carried out by a sensory motor system comprising multiple sensors (visual,auditory, and proprioceptive),multiple actuators (muscles acting on the skeletal system),and an intermediary processor that can be summarized as a multiple-input-multiple-output nonlinear dynamic time varying control system. This grand control system is capable of responding with remarkable accuracy,speed, appropriateness,versatility, and adaptability to a wide spectrum of continuous and discrete stimuli and conditions and is certainly orders of magnitude more complex and sophisticated than the most advanced robotic systems currently available. In the last decades,a great deal of research has been carried out in the fields of functional evaluation of human performance and rehabilitation engineering. These fields combine knowledge, concepts, and methods from across many disciplines (e.g., biomechanics,neuroscience, and physiology), with the aim of developing apparatuses and methods fort he measurement and analysis of complex sensory motor performance and the ultimate goal of enhancing the execution of different tasks in both healthy people and persons with reduced capabilities from different causes (injury, disease, amputation,and neural degeneration). PMID- 21642031 TI - Biosensors and molecular imaging. AB - The increase in the understanding of the physical and functional properties of the biological material, from the cellular level down to single molecules, owes its success to the development of suitable high-sensitivity platforms to image the biomaterial and analyze its response to specific stimuli. Imaging has indeed reached molecular capabilities, thanks to optical or magnetic markers [1], to the atomic force microscopy (AFM) in surface reconstruction [2], and is nearing success in three-dimensional (3-D) reconstruction thanks to X-ray holography [3]. PMID- 21642032 TI - Biomedical signal and image processing. AB - Generally, physiological modeling and biomedical signal processing constitute two important paradigms of biomedical engineering (BME): their fundamental concepts are taught starting from undergraduate studies and are more completely dealt with in the last years of graduate curricula, as well as in Ph.D. courses. Traditionally, these two cultural aspects were separated, with the first one more oriented to physiological issues and how to model them and the second one more dedicated to the development of processing tools or algorithms to enhance useful information from clinical data. A practical consequence was that those who did models did not do signal processing and vice versa. However, in recent years,the need for closer integration between signal processing and modeling of the relevant biological systems emerged very clearly [1], [2]. This is not only true for training purposes(i.e., to properly prepare the new professional members of BME) but also for the development of newly conceived research projects in which the integration between biomedical signal and image processing (BSIP) and modeling plays a crucial role. Just to give simple examples, topics such as brain computer machine or interfaces,neuroengineering, nonlinear dynamical analysis of the cardiovascular (CV) system,integration of sensory-motor characteristics aimed at the building of advanced prostheses and rehabilitation tools, and wearable devices for vital sign monitoring and others do require an intelligent fusion of modeling and signal processing competences that are certainly peculiar of our discipline of BME. PMID- 21642033 TI - Medical robotics. AB - Information and communication technology (ICT) and mechatronics play a basic role in medical robotics and computer-aided therapy. In the last three decades, in fact, ICT technology has strongly entered the health-care field, bringing in new techniques to support therapy and rehabilitation. In this frame, medical robotics is an expansion of the service and professional robotics as well as other technologies, as surgical navigation has been introduced especially in minimally invasive surgery. Localization systems also provide treatments in radiotherapy and radiosurgery with high precision. Virtual or augmented reality plays a role for both surgical training and planning and for safe rehabilitation in the first stage of the recovery from neurological diseases. Also, in the chronic phase of motor diseases, robotics helps with special assistive devices and prostheses. Although, in the past, the actual need and advantage of navigation, localization, and robotics in surgery and therapy has been in doubt, today, the availability of better hardware (e.g., microrobots) and more sophisticated algorithms(e.g., machine learning and other cognitive approaches)has largely increased the field of applications of these technologies,making it more likely that, in the near future, their presence will be dramatically increased, taking advantage of the generational change of the end users and the increasing request of quality in health-care delivery and management. PMID- 21642034 TI - Telemedicine and e-health. AB - We begin this article with the celebration of the 20th anniversary mentioning some activity lines in the area of telemedicine and e-health currently underway not only at the bioengineering department but also at other departments of the Politecnico di Milano. PMID- 21642035 TI - Integrated multiscale modeling of the nervous system: predicting changes in hippocampal network activity by a positive AMPA receptor modulator. AB - One of the fundamental characteristics of the brain is its hierarchical organization. Scales in both space and time that must be considered when integrating across hierarchies of the nervous system are sufficiently great as to have impeded the development of routine multilevel modeling methodologies. Complex molecular interactions at the level of receptors and channels regulate activity at the level of neurons; interactions between multiple populations of neurons ultimately give rise to complex neural systems function and behavior. This spatial complexity takes place in the context of a composite temporal integration of multiple, different events unfolding at the millisecond, second, minute, hour, and longer time scales. In this study, we present a multiscale modeling methodology that integrates synaptic models into single neuron, and multineuron, network models. We have applied this approach to the specific problem of how changes at the level of kinetic parameters of a receptor-channel model are translated into changes in the temporal firing pattern of a single neuron, and ultimately, changes in the spatiotemporal activity of a network of neurons. These results demonstrate how this powerful methodology can be applied to understand the effects of a given local process within multiple hierarchical levels of the nervous system. PMID- 21642036 TI - Microwave cancer imaging exploiting magnetic nanoparticles as contrast agent. AB - In this paper, a microwave technique for breast cancer imaging is presented. The approach is based on the use of magnetic nanoparticles as contrast agent to induce a nonnull magnetic contrast selectively localized within the tumor. This allows us to face cancer imaging as the reconstruction of a magnetic contrast from the corresponding scattered field. To extract, from the measured data the contribution due to the magnetic contrast, i.e., the signal meaningful for cancer imaging, the approach exploits the possibility of modulating the magnetic response of magnetic nanoparticles by means of a polarizing magnetic field. The achievable reconstruction capabilities and the robustness against uncertainties on the electric features of the surrounding electric scenario are assessed by means of numerical examples. PMID- 21642037 TI - Characterization of the electrochemical behavior of gastrointestinal fluids using a multielectrode sensor probe. AB - A characterization of gastrointestinal fluids has been performed by means of an electrochemical sensor that has potential for clinical in vivo and in vitro monitoring applications. The sensor comprised a three-electrode cell with a counter, reference, and four working electrodes, Au, Pt, Ir, and Rh. Cyclic voltammetry was used to obtain chemical information from faecal water (in vitro) and gut model (in vivo) fluids. Stable voltammetric responses were obtained for both fluids at these noble metal working electrodes. The responses differed in shape that demonstrated the discrimination capability and the potential for practical use as a tool for gastrointestinal fluid investigation. The analysis of the stability profiles in faecal water over a 14-h duration has indicated a possible adsorption mechanism with the formation of a biolayer on the sensor surface. The stability in gut model fluids over a 42-h duration has demonstrated a more stable profile, but the mechanisms involved are more complicated to determine. PMID- 21642038 TI - Common spatio-temporal pattern for single-trial detection of event-related potential in rapid serial visual presentation triage. AB - Searching for target images in large volume imagery is a challenging problem and the rapid serial visual presentation (RSVP) triage is potentially a promising solution to the problem. RSVP triage is essentially a cortically-coupled computer vision technique that relies on single-trial detection of event-related potentials (ERP). In RSVP triage, images are shown to a subject in a rapid serial sequence. When a target image is seen by the subject, unique ERP characterized by P300 are elicited. Thus, in RSVP triage, accurate detection of such distinct ERP allows for fast searching of target images in large volume imagery. The accuracy of the distinct ERP detection in RSVP triage depends on the feature extraction method, for which the common spatial pattern analysis (CSP) was used with limited success. This paper presents a novel feature extraction method, termed common spatio-temporal pattern (CSTP), which is critical for robust single-trial detection of ERP. Unlike the conventional CSP, whereby only spatial patterns of ERP are considered, the present proposed method exploits spatial and temporal patterns of ERP separately, providing complementary spatial and temporal features for high accurate single-trial ERP detection. Numerical study using data collected from 20 subjects in RSVP triage experiments demonstrates that the proposed method offers significant performance improvement over the conventional CSP method (corrected p-value < 0.05, Pearson r = 0.64) and other competing methods in the literature. This paper further shows that the main idea of CSTP can be easily applied to other methods. PMID- 21642039 TI - Volume-based features for detection of bladder wall abnormal regions via MR cystography. AB - This paper proposes a framework for detecting the suspected abnormal region of the bladder wall via magnetic resonance (MR) cystography. Volume-based features are used. First, the bladder wall is divided into several layers, based on which a path from each voxel on the inner border to the outer border is found. By using the path length to measure the wall thickness and a bent rate (BR) term to measure the geometry property of the voxels on the inner border, the seed voxels representing the abnormalities on the inner border are determined. Then, by tracing the path from each seed, a weighted BR term is constructed to determine the suspected voxels, which are on the path and inside the bladder wall. All the suspected voxels are grouped together for the abnormal region. This work is significantly different from most of the previous computer-aided bladder tumor detection reports on two aspects. First of all, the T (1)-weighted MR images are used which give better image contrast and texture information for the bladder wall, comparing with the computed tomography images. Second, while most previous reports detected the abnormalities and indicated them on the reconstructed 3-D bladder model by surface rendering, we further determine the possible region of the abnormality inside the bladder wall. This study aims at a noninvasive procedure for bladder tumor detection and abnormal region delineation, which has the potential for further clinical analysis such as the invasion depth of the tumor and virtual cystoscopy diagnosis. Five datasets including two patients and three volunteers were used to test the presented method, all the tumors were detected by the method, and the overlap rates of the regions delineated by the computer against the experts were measured. The results demonstrated the potential of the method for detecting bladder wall abnormal regions via MR cystography. PMID- 21642040 TI - Prediction based collaborative trackers (PCT): a robust and accurate approach toward 3D medical object tracking. AB - Robust and fast 3D tracking of deformable objects, such as heart, is a challenging task because of the relatively low image contrast and speed requirement. Many existing 2D algorithms might not be directly applied on the 3D tracking problem. The 3D tracking performance is limited due to dramatically increased data size, landmarks ambiguity, signal drop-out or complex nonrigid deformation. In this paper, we present a robust, fast, and accurate 3D tracking algorithm: prediction based collaborative trackers (PCT). A novel one-step forward prediction is introduced to generate the motion prior using motion manifold learning. Collaborative trackers are introduced to achieve both temporal consistency and failure recovery. Compared with tracking by detection and 3D optical flow, PCT provides the best results. The new tracking algorithm is completely automatic and computationally efficient. It requires less than 1.5 s to process a 3D volume which contains millions of voxels. In order to demonstrate the generality of PCT, the tracker is fully tested on three large clinical datasets for three 3D heart tracking problems with two different imaging modalities: endocardium tracking of the left ventricle (67 sequences, 1134 3D volumetric echocardiography data), dense tracking in the myocardial regions between the epicardium and endocardium of the left ventricle (503 sequences, roughly 9000 3D volumetric echocardiography data), and whole heart four chambers tracking (20 sequences, 200 cardiac 3D volumetric CT data). Our datasets are much larger than most studies reported in the literature and we achieve very accurate tracking results compared with human experts' annotations and recent literature. PMID- 21642041 TI - Adaptive neural network decentralized backstepping output-feedback control for nonlinear large-scale systems with time delays. AB - In this paper, two adaptive neural network (NN) decentralized output feedback control approaches are proposed for a class of uncertain nonlinear large-scale systems with immeasurable states and unknown time delays. Using NNs to approximate the unknown nonlinear functions, an NN state observer is designed to estimate the immeasurable states. By combining the adaptive backstepping technique with decentralized control design principle, an adaptive NN decentralized output feedback control approach is developed. In order to overcome the problem of "explosion of complexity" inherent in the proposed control approach, the dynamic surface control (DSC) technique is introduced into the first adaptive NN decentralized control scheme, and a simplified adaptive NN decentralized output feedback DSC approach is developed. It is proved that the two proposed control approaches can guarantee that all the signals of the closed loop system are semi-globally uniformly ultimately bounded, and the observer errors and the tracking errors converge to a small neighborhood of the origin. Simulation results are provided to show the effectiveness of the proposed approaches. PMID- 21642042 TI - Evidence-driven image interpretation by combining implicit and explicit knowledge in a Bayesian network. AB - Computer vision techniques have made considerable progress in recognizing object categories by learning models that normally rely on a set of discriminative features. However, in contrast to human perception that makes extensive use of logic-based rules, these models fail to benefit from knowledge that is explicitly provided. In this paper, we propose a framework that can perform knowledge assisted analysis of visual content. We use ontologies to model the domain knowledge and a set of conditional probabilities to model the application context. Then, a Bayesian network is used for integrating statistical and explicit knowledge and performing hypothesis testing using evidence-driven probabilistic inference. In addition, we propose the use of a focus-of-attention (FoA) mechanism that is based on the mutual information between concepts. This mechanism selects the most prominent hypotheses to be verified/tested by the BN, hence removing the need to exhaustively test all possible combinations of the hypotheses set. We experimentally evaluate our framework using content from three domains and for the following three tasks: 1) image categorization; 2) localized region labeling; and 3) weak annotation of video shot keyframes. The results obtained demonstrate the improvement in performance compared to a set of baseline concept classifiers that are not aware of any context or domain knowledge. Finally, we also demonstrate the ability of the proposed FoA mechanism to significantly reduce the computational cost of visual inference while obtaining results comparable to the exhaustive case. PMID- 21642043 TI - Prediction of the timing and the rhythm of the parkinsonian subthalamic nucleus neural spikes using the local field potentials. AB - In this paper, we discuss the use of a nonlinear cascade model to predict the subthalamic nucleus spike activity from the local field potentials recorded in the motor area of the nucleus of Parkinson's disease patients undergoing deep brain stimulation. We use a segment of appropriately selected and processed data recorded from five nuclei to acquire the information of the spike timing and rhythm of a single neuron and estimate the model parameters. We then use the rest of each recording to assess the model's accuracy in predicting spike timing, rhythm, and interspike intervals. We show that the cumulative distribution function (CDF) of the predicted spikes remains inside the 95% confidence interval of the CDF of the recorded spikes. By training the model appropriately, we prove its ability to provide quite accurate predictions for multiple-neuron recordings as well, and we establish its validity as a simple yet biologically plausible model of the intranuclear spike activity recorded from Parkinson's disease patients. PMID- 21642044 TI - Occupational exposure to asbestos and ovarian cancer: a meta-analysis. AB - OBJECTIVE: A recent Monographs Working Group of the International Agency for Research on Cancer (IARC) concluded that there is sufficient evidence for a causal association between exposure to asbestos and ovarian cancer. We performed a meta-analysis to quantitatively evaluate this association. DATA SOURCES: Searches of PubMed and unpublished data yielded a total of 18 cohort studies of women occupationally exposed to asbestos. DATA EXTRACTION: Two authors independently abstracted data; any disagreement was resolved by consulting a third reviewer. DATA SYNTHESIS: All but one study reported standardized mortality ratios (SMRs) comparing observed numbers of deaths with expected numbers for the general population; the exception was a study that reported standardized incidence ratios. For simplicity, we refer to all effect estimates as SMRs. The overall pooled SMR estimate for ovarian cancer was 1.77 (95% confidence interval, 1.37-2.28), with a moderate degree of heterogeneity among the studies (I2 = 35.3%, p = 0.061). Effect estimates were stronger for cohorts compensated for asbestosis, cohorts with estimated lung cancer SMRs > 2.0, and studies conducted in Europe compared with other geographic regions. Effect estimates were similar for studies with and without pathologic confirmation, and we found no evidence of publication bias (Egger's test p-value = 0.162). CONCLUSIONS: Our study supports the IARC conclusion that exposure to asbestos is associated with increased risk of ovarian cancer. PMID- 21642045 TI - Acute respiratory inflammation in children and black carbon in ambient air before and during the 2008 Beijing Olympics. AB - BACKGROUND: Epidemiologic evidence for a causative association between black carbon (BC) and health outcomes is limited. OBJECTIVES: We estimated associations and exposure-response relationships between acute respiratory inflammation in schoolchildren and concentrations of BC and particulate matter with an aerodynamic diameter of <= 2.5 MUm (PM2.5) in ambient air before and during the air pollution intervention for the 2008 Beijing Olympics. METHODS: We measured exhaled nitric oxide (eNO) as an acute respiratory inflammation biomarker and hourly mean air pollutant concentrations to estimate BC and PM2.5 exposure. We used 1,581 valid observations of 36 subjects over five visits in 2 years to estimate associations of eNO with BC and PM2.5 according to generalized estimating equations with polynomial distributed-lag models, controlling for body mass index, asthma, temperature, and relative humidity. We also assessed the relative importance of BC and PM2.5 with two-pollutant models. RESULTS: Air pollution concentrations and eNO were clearly lower during the 2008 Olympics. BC and PM2.5 concentrations averaged over 0-24 hr were strongly associated with eNO, which increased by 16.6% [95% confidence interval (CI), 14.1-19.2%] and 18.7% (95% CI, 15.0-22.5%) per interquartile range (IQR) increase in BC (4.0 MUg/m3) and PM2.5 (149 MUg/m3), respectively. In the two-pollutant model, estimated effects of BC were robust, but associations between PM2.5 and eNO decreased with adjustment for BC. We found that eNO was associated with IQR increases in hourly BC concentrations up to 10 hr after exposure, consistent with effects primarily in the first hours after exposure. CONCLUSIONS: Recent exposure to BC was associated with acute respiratory inflammation in schoolchildren in Beijing. Lower air pollution levels during the 2008 Olympics also were associated with reduced eNO. PMID- 21642046 TI - Social disparities in nitrate-contaminated drinking water in California's San Joaquin Valley. AB - BACKGROUND: Research on drinking water in the United States has rarely examined disproportionate exposures to contaminants faced by low-income and minority communities. This study analyzes the relationship between nitrate concentrations in community water systems (CWSs) and the racial/ethnic and socioeconomic characteristics of customers. OBJECTIVES: We hypothesized that CWSs in California's San Joaquin Valley that serve a higher proportion of minority or residents of lower socioeconomic status have higher nitrate levels and that these disparities are greater among smaller drinking water systems. METHODS: We used water quality monitoring data sets (1999-2001) to estimate nitrate levels in CWSs, and source location and census block group data to estimate customer demographics. Our linear regression model included 327 CWSs and reported robust standard errors clustered at the CWS level. Our adjusted model controlled for demographics and water system characteristics and stratified by CWS size. RESULTS: Percent Latino was associated with a 0.04-mg nitrate-ion (NO3)/L increase in a CWS's estimated NO3 concentration [95% confidence interval (CI), 0.08 to 0.16], and rate of home ownership was associated with a 0.16-mg NO3/L decrease (95% CI, -0.32 to 0.002). Among smaller systems, the percentage of Latinos and of homeownership was associated with an estimated increase of 0.44 mg NO3/L (95% CI, 0.03-0.84) and a decrease of 0.15 mg NO3/L (95% CI, -0.64 to 0.33), respectively. CONCLUSIONS: Our findings suggest that in smaller water systems, CWSs serving larger percentages of Latinos and renters receive drinking water with higher nitrate levels. This suggests an environmental inequity in drinking water quality. PMID- 21642047 TI - Comparison of serum bisphenol A concentrations in mice exposed to bisphenol A through the diet versus oral bolus exposure. AB - BACKGROUND: Bisphenol A (BPA) is a widely produced endocrine-disrupting chemical. Diet is a primary route of exposure, but internal exposure (serum concentrations) in animals and humans has been measured only after single oral bolus administration. OBJECTIVE: We compared serum concentrations of BPA over a 24-hr period after oral bolus administration or ad libitum feeding in mice and assessed for buildup with dietary exposure. METHODS: Adult female mice were administered [dimethyl-d6]-BPA (BPA-d6) as a single oral bolus (20 mg/kg body weight) or fed a diet containing 100 mg BPA-d6/kg feed weight ad libitum for 1 week. Serum concentrations were analyzed using isotope dilution liquid chromatography coupled with electrospray tandem mass spectrometry and compared between exposure groups over the first 23 hr and after 7 days of dietary exposure. RESULTS: Maximum concentration (Cmax) for BPA-d6 during the first 24 hr was reached at 1 hr and 6 hr for oral bolus and diet groups, respectively. Relative BPA-d6 bioavailability (unconjugated BPA-d6) was higher in diet-exposed mice than in the bolus group despite a relative lower absorption, a phenomenon consistent with an inhibitory effect of food on first-pass hepatic metabolism. In mice with ongoing dietary exposure, unconjugated BPA-d6 was higher on day 7 than on day 1. CONCLUSIONS: This is the first report of serum BPA concentrations in an animal model exposed to this chemical via the diet. Although bolus administration of BPA-d6 led to peak concentrations within 1 hr, Cmax for diet-exposed mice was delayed for several hours. However, bolus administration underestimates bioavailable serum BPA concentrations in animals-and presumably humans-than would result from dietary exposure. Exposure via diet is a more natural continuous exposure route than oral bolus exposure and is thus a better predictor of BPA concentrations in chronically exposed animals and humans. PMID- 21642048 TI - Acute pesticide illnesses associated with off-target pesticide drift from agricultural applications: 11 States, 1998-2006. AB - BACKGROUND: Pesticides are widely used in agriculture, and off-target pesticide drift exposes workers and the public to harmful chemicals. OBJECTIVE: We estimated the incidence of acute illnesses from pesticide drift from outdoor agricultural applications and characterized drift exposure and illnesses. METHODS: Data were obtained from the National Institute for Occupational Safety and Health's Sentinel Event Notification System for Occupational Risks-Pesticides program and the California Department of Pesticide Regulation. Drift included off target movement of pesticide spray, volatiles, and contaminated dust. Acute illness cases were characterized by demographics, pesticide and application variables, health effects, and contributing factors. RESULTS: From 1998 through 2006, we identified 2,945 cases associated with agricultural pesticide drift from 11 states. Our findings indicate that 47% were exposed at work, 92% experienced low-severity illness, and 14% were children (< 15 years). The annual incidence ranged from 1.39 to 5.32 per million persons over the 9-year period. The overall incidence (in million person-years) was 114.3 for agricultural workers, 0.79 for other workers, 1.56 for nonoccupational cases, and 42.2 for residents in five agriculture-intensive counties in California. Soil applications with fumigants were responsible for the largest percentage (45%) of cases. Aerial applications accounted for 24% of cases. Common factors contributing to drift cases included weather conditions, improper seal of the fumigation site, and applicator carelessness near nontarget areas. CONCLUSIONS: Agricultural workers and residents in agricultural regions had the highest rate of pesticide poisoning from drift exposure, and soil fumigations were a major hazard, causing large drift incidents. Our findings highlight areas where interventions to reduce off target drift could be focused. PMID- 21642049 TI - [Economic impact of transarterial chemoembolization with drug eluting beads in the treatment of hepatocellular carcinoma]. AB - Transarterial chemoembolization (TACE) is the standard treatment in patients with unresectable non-metastatic hepatocellular carcinoma (HCC). New drug eluting beads aim at improving efficacy of TACE in retaining as long as possible the anticancer drug within the tumor. Our monocentric study compares direct hospital medical costs, according to two different methods, for a first course of conventional TACE and for a first course of TACE using drug eluting beads in 30 patients with HCC. The average cost of a first course of conventional TACE valued by the analytic accounting system is 4 332 ? versus 3 577 ? for a first course of TACE using drug eluting beads. The average cost of a first course of conventional TACE valued by the official tariffs from the new French Diagnosis Related Group prospective payment system is 4 507 ? (+175 ?) versus 2 852 ? (-725 ?) for a first course of TACE using drug eluting beads. Our study shows that a first TACE using drug eluting beads, valued by the official tariffs from the new French Diagnosis Related Group prospective payment system, is significantly (p = 0.006) less expensive than a first conventional TACE. PMID- 21642050 TI - [New anticoagulants in the prevention and treatment of venous thromboembolism]. AB - Clinical data on the risk factors, incidence, consequences and current treatment options of venous thromboembolism are reviewed. Current guidelines advise anticoagulant treatment for a few weeks or months in immobilized patients treated in hospital, and after major surgery. The initial treatment is based on heparin, followed by vitamin K antagonist treatment. Recently a number of new, partially orally administered medications have undergone clinical investigations and based on the results three of them were also registered for the prevention and treatment of venous thromboembolism. Direct thrombin inhibitors, direct and indirect Factor Xa inhibitors exhibited proven non-inferiority or superiority compared with traditional treatment options. The superior efficacy or non inferiority was not accompanied with an increase in the bleeding risk. Results of the most important clinical trials are reviewed. Based on these results, prevention and treatment of venous thromboembolism will change substantially in the near future. PMID- 21642051 TI - [Functional vascular alterations associated with aortic valve stenosis]. AB - Degenerative changes, atherosclerotic process and calcification of valvular leaflets are mostly responsible for valvular aortic valve stenosis, but congenital bicuspid aortic valve and rheumatic fever in history are also known predisposing factors. Aortic valve stenosis is frequently associated with different functional vascular alterations. The aim of this review is to demonstrate these vascular alterations evaluated by non-invasive methods and underlying physiologic and pathophysiologic processes. PMID- 21642052 TI - [Epidemiology of malignant melanoma (Clinical experience at the National Institute of Oncology in Hungary)]. AB - The National Cancer Registry, which has been running since 1999 in Hungary, supplies more and more precise data about the growing incidence of malignant melanoma although nationwide melanoma database currently does not exist. As in the National Institute of Oncology a notable percentage of patients (20-25%) were treated with newly diagnosed melanoma, conclusions may be drawn for the efficacy of primary prevention for the whole country. METHODS: The recent study compares the data of patients presented in the Institute with cutaneous malignant melanoma in 1998 and ten years later, in 2008. The histology parameters of tumors were also analyzed. The two groups were compared according to age, gender, localization and histology parameters (histological types Breslow and Clark numbers). In case of continuous variables the Mann-Whitney test were used to determine significant differences. Categorical variables were checked by chi2 test. RESULTS: 149 and 377 cutaneous melanomas were diagnosed in 1998 and in 2008, respectively, which accounts for 153% increase in ten years. The mean age of patients was 56.3 and 57.2 years in 1998 and in 2008, respectively. The per cent of males was 43% in 1998 and 49% in 2008. Trunk was the most frequent localization in both years: 39% and 46%, then lower limbs (28% and 22%), upper limbs (21% and 18%) and the head and neck region (12% and 14%). Superficial melanoma was found the most frequent histological type (52 and 54%) followed by nodular melanoma (31% and 23%). Rate of in situ melanoma changed from 10% to 15%. The mean of Breslow numbers was 2.2 mm in 1998 and 1.6 mm in 2008, the difference was highly significant (p = 0.0002). Clark numbers were also decreasing, although the difference was not significant (p = 0.08). The majority of patients were presented with Clark III depth melanoma in both years (38% and 32%). CONCLUSIONS: It seems that forces emphasizing the importance of early recognition in medical education and in media are not futile considering that principally the early diagnosis can improve the melanoma's prognosis - which is represented mainly by Breslow tumor depth. PMID- 21642053 TI - [Clinical significance and the first identification of human parechoviruses in Hungary]. AB - Human parechoviruses (HPeV) belonging to the family Picornaviridae are widespread enteric pathogens and are associated with various clinical syndromes in human. At present, 16 HPeV genotypes (HPeV1-16) are known. There is no report on the detection of HPeVs in Central Europe. AIMS: The aim of the retrospective study was to detect and characterize HPeVs using molecular methods in cell cultures with "enterovirus-like" cytophatic effect (CPE) archived between 1990 and 2004, in two virology laboratories, in Hungary. MATERIALS AND METHODS: In Laboratory I, fecal samples from children with symptoms of gastroenteritis under the age of 10 years were cultured as a previous routine diagnostic laboratory protocol for "enterovirus". Cell cultures indicating CPE were archived between 1990 and 2000. In Laboratory II, 2 fecal samples, a liquor and a nasopharyngeal aspirate were re tested which contained an "enterovirus-like" virus in cell cultures and were positive by HPeV1 neutralization immunosera between 2000 and 2004. Specimens were tested retrospectively for HPeV by reverse transcription-PCR (RT-PCR) method using 5'UTR conserved primers. Specific primers were designed to determine the HPeV structural region (VP0-VP3-VP1). RESULTS: 9 of the 66 archived samples (9.1%) from Laboratory I and all the 4 samples from Laboratory II were found to be HPeV-positive. 10 samples were identified as HPeV1, 2 were HPeV4 and 1 could not be determined. 3 HPeV1 clusters were identified in Laboratory I according to the isolation date originated from years 1990/1991, 1992/1995 and 1998. HPeV1 was detected in clinical syndromes: gastroenteritis (in a 24-years-old adult), recurrent stomatitis aphtosa (in a 42-years-old adult), encephalitis and ataxia cerebellaris acuta in infants and children in Laboratory II. CONCLUSIONS: This is the first detection of HPeVs in Central Europe. Detection and genetic characterization of HPeV in available historical samples infected with previously unidentifiable agents with "enterovirus-like" cytopathogenic effect may help to understand the clinical importance and spectrum of the infections and the genetic diversity and evolution of these viruses. PMID- 21642054 TI - [Endometriosis]. PMID- 21642056 TI - Poststroke hypertonicity: upper limb assessment and treatment. AB - Hypertonicity is common in patients with upper limb dysfunction following hemiplegic stroke and is associated with greater impairment, worse function, and lower health-related quality of life. In addition to increased rest activity, abnormal patterns of muscle activation, such as spastic co-contraction, may contribute to disability. In the upper limb, flexor muscles are more commonly involved distally, and at the shoulder, spasticity of adductors, flexors, and internal rotators is most often observed. Prior to interventions, a history regarding prior interventions, comorbid diagnoses, and limitations imposed by abnormal tone should be elicited. Commonly used scales to assess hypertonicity include the Modified Ashworth, the Modified Tardieu, the Spasm Frequency, the Disability Assessment, the Fugl-Meyer, and the Motor Assessment Scales. Treatment interventions for upper limb hypertonicity include stretching, splinting, strengthening of antagonist muscles, oral medications, and focal injections (phenol or botulinum toxins). Intrathecal baclofen may also impact upper limb tone. For focal injections, correct identification of muscles contributing to problematic tone is evaluated by eliciting resistance to movement at rest and observation of patterns of tightness as the limb is used functionally. The botulinum toxins have been shown to decrease tone in stroke survivors and improve active and passive functioning. Because secondary changes such as contractures and weakness may occur with prolonged hypertonicity, therapy to improve range of motion, strengthen weakened muscles, and incorporate use of the limb should be considered following focal injections, oral medications, or intrathecal pump placement. PMID- 21642057 TI - The underutilization of intrathecal baclofen in poststroke spasticity. AB - Stroke is one of the leading causes of adult disability in the United States, with a reported prevalence of 6.4 million people. Spasticity is one of the clinical features of the upper motor neuron syndrome seen after a stroke. The prevalence of spasticity after a stroke ranges from 17% to 42.6%, and an average of two-thirds of people with spasticity have upper and lower extremity involvement. Oral medications and botulinum neurotoxin injections are current treatments for problematic spasticity. However, these treatments are often limited by side effects or dose ceilings. Intrathecal baclofen (ITB) is a proven method for the management of disabling spasticity from multiple etiologies. Studies have demonstrated improved mobility, activities of daily living, and quality of life in spastic poststroke patients. Despite the benefits of ITB, fewer than 1% of stroke patients with severe disabling spasticity are being treated with ITB. This article will review the prevalence of severe poststroke spasticity and the rate of ITB use and will discuss reasons for its limited use in stroke survivors. PMID- 21642058 TI - Rehabilitation techniques to maximize spasticity management. AB - Improvement in functional skills is typically a goal of spasticity management. Spasticity management alone will improve the positive signs of the upper motor neuron syndrome without functional change. In this review, we demonstrate that a variety of therapy modalities are required to facilitate these improvements and impact the negative signs of the upper motor neuron syndrome. The evidence for neuromuscular electrical stimulation, surface electromyography training, serial casting, body weight-supported treadmill training, constraint-induced movement therapy, strengthening, and endurance training is reviewed as it relates to spasticity management. PMID- 21642059 TI - A novel adaptive mixed reality system for stroke rehabilitation: principles, proof of concept, and preliminary application in 2 patients. AB - This article presents the principles of an adaptive mixed reality rehabilitation (AMRR) system, as well as the training process and results from 2 stroke survivors who received AMRR therapy, to illustrate how the system can be used in the clinic. The AMRR system integrates traditional rehabilitation practices with state-of-the-art computational and motion capture technologies to create an engaging environment to train reaching movements. The system provides real-time, intuitive, and integrated audio and visual feedback (based on detailed kinematic data) representative of goal accomplishment, activity performance, and body function during a reaching task. The AMRR system also provides a quantitative kinematic evaluation that measures the deviation of the stroke survivor's movement from an idealized, unimpaired movement. The therapist, using the quantitative measure and knowledge and observations, can adapt the feedback and physical environment of the AMRR system throughout therapy to address each participant's individual impairments and progress. Individualized training plans, kinematic improvements measured over the entire therapy period, and the changes in relevant clinical scales and kinematic movement attributes before and after the month-long therapy are presented for 2 participants. The substantial improvements made by both participants after AMRR therapy demonstrate that this system has the potential to considerably enhance the recovery of stroke survivors with varying impairments for both kinematic improvements and functional ability. PMID- 21642060 TI - Short- and long-term effects of an inhibitor hand splint in poststroke patients: a randomized controlled trial. AB - PURPOSE: To evaluate the effects of a hand splint with reflex inhibitory characteristics on balance and functional ambulation activities by means of a randomly allocated controlled study. METHODS: Nineteen chronic stroke patients were randomly allocated to the control (n = 9) and study (n = 10) groups to compare the immediate and long-term effects of a splint with reflex inhibitory characteristics. The patients in the splinted group were asked to wear their splints at least 2 hours each day, during ambulation or when they felt the need. Subjects were evaluated initially and after 2, 4, and 6 months of splint usage with the Berg Balance Scale, Functional Reach test (FR), Timed Up & Go test (TUG), and L test. The same tests were used to evaluate the control group. RESULTS: Timed within-group assessments showed no difference in the control group across the time interval. A favorable difference was detected in some of the evaluations for FR, TUG, and L test in the study group. Intergroup comparisons showed that the only difference between the groups was seen for TUG values at the fourth assessment in favor of the study group. Qualitative evaluations showed that the patients were compliant and generally satisfied with their splints. CONCLUSION: Hand splints with reflex inhibitory characteristics have no significant effect on balance and functional ambulation activities in chronic poststroke patients. Further studies investigating their effect on pain and associated reactions in this patient population are warranted. PMID- 21642061 TI - Dual-task demands of hand movements for adults with stroke: a pilot study. AB - For adults with stroke, walking while performing a cognitive task can be challenging, resulting in slower walking, poorer cognitive performance, or decreased performance on both tasks. It is not known if dual-task deficits are also present for upper limb movements for adults with stroke. PURPOSE: To determine if unilateral movements of the affected and less affected hand are compromised when walking or talking. METHODS: Nineteen community-dwelling adults with stroke were video- and audiotaped while performing in single- and dual-task conditions. Tasks included repeated, rhythmic hand movements with the affected and less affected hand, walking a narrow pathway, and speaking. For dual-task conditions, movements of each hand were done while walking and while talking. The rate of hand movement, cadence, and speech rate were analyzed using repeated measures analyses of variance. RESULTS: Affected hand movement rate was the same for single- and dual-task conditions. The rate of less affected hand movement was affected by dual-task conditions; this was due to an increase in hand movement rate while talking. Examination of cadence and speech rates revealed that cadence was decreased when moving the affected hand. Speech rate increased when accompanied by hand movements, but post hoc analyses were not significant. CONCLUSION: For those with stroke, dual-task deficits are seen with slower walking while moving the affected hand. In contrast, hand movements while speaking may have a more complex relationship, with possible faster speech rates in dual-task conditions. PMID- 21642062 TI - Recovery of upper extremity motor function post stroke with regard to eligibility for constraint-induced movement therapy. AB - PURPOSE: To examine eligibility for modalities such as constraint-induced movement therapy (CIMT) and modified CIMT (mCIMT) in the subacute phase after stroke and to define the share of patients who should be offered this treatment. METHODS: A prospective, repeated-measures design was used. A total of 100 consecutive patients with arm paresis 1 to 2 weeks post stroke were screened. Eligible for CIMT were patients who were cognitively intact, medically stable, and able to extend the wrist and 3 fingers 10 degrees as a lower limit. The active range of motion was registered, and motor function was assessed by the Action Research Arm Test (ARAT) and the Nine Hole Peg Test at 1 to 2 weeks, 4 weeks, and 3 months post stroke. RESULTS: From 100 patients, 54 were excluded from motor assessment, mostly due to cognitive impairments. Of the remaining 46 patients, 21 (46%) were eligible according to motor function of the hand at 1 to 2 weeks post stroke, whereas in the other patients motor function was either too good or too poor. The share of patients eligible declined to 31% after 4 weeks and 15% after 3 months. Within 3 months, 60% reached reasonable dexterity, expressed by an ARAT score ? 51, all receiving standard rehabilitation. CONCLUSION: Results indicate that eligibility for CIMT or mCIMT should not be considered before 4 weeks post stroke because much improvement in arm function was shown to occur during the first month post stroke with standard rehabilitation. PMID- 21642063 TI - Effects of limb loading on gait initiation in persons with moderate hemiparesis. AB - PURPOSE: To examine the immediate effect of symmetrical weight bearing (SWB) on temporal events of gait initiation (GI) patterns and timing and amplitude of lower distal limb muscles activity during GI in persons with hemiparesis. METHOD: The study was a within-subjects design. Twelve persons with hemiparesis were recruited from the Veterans Affairs Brain Rehabilitation Research Center at the Malcom Randall Veterans Affairs, Gainesville, Florida. GI trials were performed from 4 beginning limb-loading conditions presented in a randomized order: (1) GI with the paretic limb during natural (asymmetrical) weight bearing (NWB); (2) GI with the nonparetic limb during NWB; (3) GI with the paretic limb during SWB; and (4) GI with the nonparetic limb during SWB. Temporal events of ground reaction forces (GRFs) and timing and amplitude of distal muscles activity were measured during GI trials in a motion analysis laboratory. RESULTS: There were no significant effects of SWB on the temporal events of GRFs and timing and amplitude of distal muscles activity when initiating gait with the paretic limb. Onset of tibilais anterior (TA) muscle was delayed significantly with less amplitude when initiating gait with the paretic limb in both NWB and SWB conditions. However, when initiating gait with the nonparetic limb, TA muscle on the paretic limb was activated normally with greater amplitude in both NWB and SWB conditions. CONCLUSION: Initiating gait with the nonparetic limb as pregait activity may more effectively challenge the dynamic balance for a symmetrical gait pattern than the standard SWB in persons with hemiparesis. PMID- 21642064 TI - Robot-assisted gait training for patients with hemiparesis due to stroke. AB - Robot-assisted devices are becoming a popular alternative to manual facilitation in stroke rehabilitation. These devices have the potential to reduce therapist burden and treatment costs; however, their effectiveness in terms of functional recovery remains in question. This pilot study compared the outcomes of a stroke rehabilitation program that incorporates robot-assisted gait training (RAGT) with a more traditional therapy program that does not. Twenty hemiparetic stroke patients were recruited at a rehabilitation hospital in Houston, Texas, and were randomly assigned to 2 groups. The control group (n = 10) received 24 1-hour sessions of conventional physical therapy, whereas the RAGT group (n = 10) received 24 1-hour sessions of conventional physical therapy combined with RAGT on a treadmill. Gait function was assessed before and after treatment by an 8-m walk test, a 3-minute walk test, and the Tinetti balance assessment. Both groups showed significant improvement in all 3 outcome measures following treatment (P < .05), but there was no difference between groups. It is concluded that RAGT may provide improvements in balance and gait comparable with conventional physical therapy. A larger multicenter trial is required to investigate the effectiveness of RAGT in hemiparetic stroke. PMID- 21642065 TI - Adherence to a home-based exercise program for individuals after stroke. AB - BACKGROUND: Although aerobic training (AT) and resistance training (RT) have been shown to improve functional abilities in patients post stroke, few patients participate, with many doing so for only a short duration. PURPOSE: To retrospectively identify factors that affect adherence to a home-based exercise program adapted for stroke patients in a cardiac rehabilitation program during and after program completion. METHODS: Fourteen participants (age 63 +/- 3 years, 37 +/- 34 months post stroke) attended the rehabilitation center on a weekly (24 weeks) and then monthly (2 months) basis. Patients were required to complete 4 AT and 1 to 2 RT sessions away from the center each week. A 16-item survey exploring adherence to home-based workouts was administered. RESULTS: Seven patients were currently participating (mean time in program, 19.4 +/- 8 weeks) and 7 had graduated (mean of 32.8 +/- 28 weeks post graduation) from the program. Current participants had higher adherence than graduated participants to AT (100% vs 76%; P < .01) and RT (100% vs 55%; P < .01). The most common factors motivating participants were to improve overall health, improve functional abilities, and enhance confidence and to reduce musculoskeletal issues. The most common factors preventing workouts were lack of motivation, musculoskeletal issues, and fatigue. There was a negative correlation between age and adherence to AT in the graduated group. CONCLUSION: Adherence to home-based exercise is superior during participation in an organized group program, with decline after graduation. PMID- 21642066 TI - How to succeed in patient-centered aphasia therapy and measure results. PMID- 21642067 TI - Residual cardiac risk reduction beyond lipid lowering. PMID- 21642068 TI - A stepwise algorithm for the surgical resection of a hyper-nephroma involving the inferior vena cava. AB - INTRODUCTION: Techniques for the resection of renal tumours (RT) with extension to the inferior vena cava (IVC) are based on the experience of individual units. We attempt to provide a logical approach to the surgical strategies in a stepwise fashion. METHODS: Over 6 years, 9 patients with RT invading the IVC underwent surgery. There were 6 males. The extension was at level IV in 3 and III in 6 cases. Cardiopulmonary bypass was used in 7 and hypothermia and circulatory arrest in 2 patients with level IV disease. The results and an algorithm of the plan of action in relation to the level of extension are presented. RESULTS: Regarding postoperative morbidity, inotropic support was needed in 5 patients, a prolonged ICU stay in 3 (33.3%), tracheostomy in 1 (11.1%). Methicillin-resistant Staphylococcus aureus infection occurred in 1, sepsis in 2, cerebrovascular accident in 1. There were 2 deaths (22.2%). For level I-II disease there was no cardiothoracic involvement. For level III we used cardiopulmonary bypass and control of the cavo-atrial junction. For level IV or suboptimal thrombectomy of level III disease, we used brief periods of circulatory arrest and repair of the cavotomy with a pericardial patch. CONCLUSIONS: Total clearance of the IVC from an adherent tumour is important for prognosis, therefore extensive level III and IV disease presents a surgical challenge. We recommend cardiopulmonary bypass for level III and brief periods of total circulatory arrest for level IV disease. PMID- 21642069 TI - Aortic valve replacement in octogenarians: outcome and predictors of complications. AB - INTRODUCTION: We aimed to evaluate the outcome in octogenarians after aortic valve replacement (AVR) and to determine the perioperative parameters that were predictive of a complicated postoperative course. METHODS: The study population included 304 patients (65% male) aged 82.7 +/- 3.5 years who underwent AVR alone (63%), in combination with coronary artery bypass grafting (28%) or with other procedures (9%), between 1998 and 1/2008. Most patients suffered from combined valve disease. RESULTS: Mechanical valves were implanted in 50% of the patients. The in-hospital mortality was 5.8%. The stay in the intensive care unit was 2.3 +/- 0.5 days and in hospital 15.3 +/- 2.6 days. After multivariate analysis we were able to identify some predictors for in-hospital mortality, such as preoperative cardiogenic shock (p<0.02), ejection fraction <0.3 (p<0.03), diameter of prosthesis <21 mm (p<0.05), and redo surgery. The most important predictors for postoperative complications after AVR were preoperative renal failure, additional surgical procedures (i.e. coronary artery bypass, mitral valve) and prolonged aortic crossclamping (all p<0.05). CONCLUSIONS: The outcome after AVR in octogenarians is satisfactory; the operative risk is acceptable and might even be reduced with an individual approach to perioperative management in high-risk patients. PMID- 21642070 TI - Modulation of hemodynamics, endogenous antioxidant enzymes, and pathophysiological changes by Angiotensin-converting enzyme inhibitors in pressure-overload rats. AB - INTRODUCTION: We sought to assess the role of angiotensin-converting enzyme (ACE) inhibitors on systolic blood pressure (BP), endogenous antioxidant enzymes and histopathological changes in pressure-overload rats. METHODS: Pressure overload was produced in male rats by abdominal aortic banding (AAB) using a blunt 22 gauge needle, as a model of cardiac hypertrophy. After surgery, AAB-induced hypertensive (AABIH) groups were treated with captopril 4 mg and ramipril 10 mg/kg per day p.o. for 16 weeks. At 16 weeks, rats were observed for general characteristics and mortality, non-invasive blood pressure (NIBP) and endogenous antioxidant enzyme catalase and superoxide dismutase (SOD) activity and histological evaluation of target organs. RESULTS: In the AABIH group a significant increase in systolic BP was observed in week 3 (149.3 +/- 0.821) and persisted until week 16, along with lower levels of serum catalase (144.7 +/- 2.204) and SOD (12.92 +/- 0.4601) activity compared to the control group. Captopril and ramipril treated groups showed a significantly smaller increase in systolic BP (25.47 +/- 3.685, 20.21 +/- 3.306) and greater serum SOD (27.33 +/- 2.338, 28.95 +/- 1.143) and catalase (181.7 +/- 8.407, 187.9 +/- 8.497) activity, respectively, than the hypertensive rats. The histological changes induced in target organs (heart, liver, kidneys and thoracic aorta) in AABIH rats were attenuated in treated rats. CONCLUSIONS: ACE-inhibition causes an improvement in myocardial antioxidant reserve, reduces oxidative stress, and prevents pathophysiological alterations, while showing a trend for potential target organ protection in hypertensive rats. PMID- 21642071 TI - Continuous infusion of furosemide combined with low-dose dopamine compared to intermittent boluses in acutely decompensated heart failure is less nephrotoxic and carries a lower readmission at thirty days. AB - INTRODUCTION: Furosemide is a potent loop diuretic that is widely used in the management of heart failure. Several reports have suggested that continuous intravenous administration of loop diuretics may be superior to intermittent administration. In addition the effect of low-dose dopamine to improve renal perfusion might be of benefit to this patient cohort. METHODS: We retrospectively evaluated 116 consecutive cardiac care unit patients, who were admitted with acute decompensated heart failure and were divided into two equal groups according to diuretic protocol. Group A patients received furosemide by continuous infusion combined with low-dose dopamine infusion. Group B patients received bolus therapy of intravenous furosemide. The effect on renal function and readmission rate was recorded. RESULTS: Among 116 patients (60% males, average age 71, range 46-96 years) 41% had ischemic cardiomyopathy, NYHA functional Class was 3.5 +/- 0.5 and average EF was 21% +/- 7%. On admission, patients in Group A had creatinine (Cr) 2.3 +/- 0.2 mg/dL, blood urea nitrogen (BUN) 49.2 +/- 25 mg/100 ml and median b-type natriuretic peptid (BNP) 1340 pg/mL, compared to group B patients with Cr 1.7 +/- 1.2 mg/dL, BUN 32 +/- 22 mg/100 ml and median BNP 1106 pg/mL. The average furosemide dose in group A was 7.9 +/- 3.5 mg/hr compared to 7.6 +/- 2.7 mg/hr for group B (p=NS). At the end of the study, patients in group A had lower Cr 1.8 +/- 0.9 (p=0.0001), lower BUN 43.6 +/- 22.9 (p=NS), an increase in estimated glomerular filtration rate 57.4 +/ 27.4, a shorter hospital stay (p=0.015) and lower readmission rates at 30 days (p=0.0003). CONCLUSIONS: Continuous infusion of furosemide in addition to low dose dopamine is safe, effective and less nephrotoxic than intermittent boluses in patients admitted with acute decompensated heart failure and portends a shorter hospital stay and lower readmission rates at 30 days. PMID- 21642072 TI - Clopidogrel resistance: current aspects and future directions. PMID- 21642073 TI - Chronic stable angina: percutaneous coronary intervention or medication? PMID- 21642074 TI - Epicardial ventricular tachycardia ablation: the last frontier in interventional electrophysiology? PMID- 21642075 TI - Embolic stroke in a patient with metastatic renal cell cancer. PMID- 21642076 TI - Mobile masses in the aortic arch in a patient with acute embolic event. AB - We describe a 45-year-old woman with peripheral embolism in whom echocardiography revealed mobile masses in the aortic arch that were characterized as atherothrombi. The masses were resected surgically 2 months later, since treatment with antiplatelets, anticoagulants and statins failed to resolve them. PMID- 21642077 TI - Successful right coronary artery drug-eluting stent implantation in a male patient with situs inversus. AB - We describe a case of successful right coronary artery angioplasty in a male subject with situs inversus. Although the interventional management of such individuals follows the standard general rules, there are also some technical aspects that should be taken into consideration in this special clinical setting. PMID- 21642078 TI - Contrast echocardiography: contribution to diagnosis of left ventricular non compaction cardiomyopathy. AB - Left ventricular non-compaction (LVNC) is a cardiomyopathy considered to be caused by arrest of normal embryogenesis of the endocardium and myocardium. Echocardiography has been the preferred diagnostic procedure; however, the correct diagnosis is often missed or delayed due to the lack of knowledge about this uncommon disease and its similarities to other diseases of the myocardium and endocardium. Here we present two cases: an asymptomatic 39-year-old man who was considered to be suffering from dilated cardiomyopathy (DCM) for four years; and an asymptomatic 19-year-old man who was considered to be suffering from hypertrophic cardiomyopathy. In a recent echocardiography study carried out in our echo lab, we recognized the morphological diagnostic criteria of LVNC. Contrast echocardiography, a low-cost, easy, repeatable, real-time, and non invasive technique with no ionising radiation, shows a high correlation in the diagnosis of LVNC. In echocardiography, the importance of contrast agents is twofold, as they can be considered essential for a reliable differentiation between the compacted and the non-compacted myocardium, while at the same time they allow accurate measurement of the ratio. Heightened clinical suspicion is necessary for the accurate diagnosis and management of diseases. PMID- 21642079 TI - A large angiosarcoma of the right atrium: anaesthetic management. AB - We present the case of a young man diagnosed with a right atrial mass and a large pericardial effusion. The patient had presented in the emergency department with chest pain, shortness of breath, pedal oedema and loss of appetite. A transthoracic echocardiogram showed a bright echodensity in the right atrium with a large pericardial effusion. He was treated for presumed tubercular pericardial effusion. Pericardiocentesis showed a straw-coloured non-tubercular pericardial effusion. Surgical removal of the right atrial tumour was planned with cardiopulmonary bypass support. The tumour could only be partially resected due to large adhesions with the myocardium. The patient suffered a cardiorespiratory arrest in the intensive care unit 3 hours after surgery due to persistent bleeding in the pericardial cavity with refractory hypovolemic shock and could not be revived. The pathological examination performed later revealed a primary cardiac angiosarcoma. The case highlights the initial clinical presentation, current diagnostic modalities, and anaesthetic management options for cardiac angiosarcoma. PMID- 21642080 TI - Congenital tricuspid valve blood cyst: imaging with three-dimensional contrast echocardiography. AB - We describe the case of an 88-year-old woman who presented in WHO functional class III and breathless on exertion. She was investigated for pulmonary hypertension. However, two-dimensional echocardiography demonstrated a rounded mass attached to the septal leaflet of the tricuspid valve. Real-time three dimensional echocardiography with contrast microbubbles was the imaging modality of choice to characterise the cyst and to guide the decision as to the best therapeutic approach for the patient. PMID- 21642081 TI - Isolated right atrial metastasis of malignant melanoma mimicking a myxoma. AB - Malignant melanoma is the malignancy with the highest propensity for cardiac metastasis. Metastasis to the heart usually occurs in the setting of disseminated disease and is therefore commonly associated with a poor prognosis. We report the case of a 51-year-old woman with a previous history of cutaneous malignant melanoma who presented with a symptomatic, isolated right atrial metastasis attached via a narrow stalk to the interatrial septum, thus resembling a myxoma. The lesion was completely resected, rendering the patient symptom and, potentially, disease-free. The case illustrates the importance of cardiac evaluation in the management of patients with melanoma. PMID- 21642082 TI - Hypertrophic cardiomyopathy: what have we learned in fifty years? PMID- 21642083 TI - The future of echocardiography. PMID- 21642084 TI - Plant biomechanics: an overview and prospectus. AB - We provide a brief overview of the articles appearing in this special issue and place them in the context of the long history of the study of plant biomechanics and what we judge to be the next major intellectual and/or technological challenges in this field. PMID- 21642085 TI - Experimental approaches used to quantify physical parameters at cellular and subcellular levels. AB - From a mechanical point of view, plant and hyphal cells are more complex than their animal counterparts because the variety of structural components determining cellular architecture is broader. In addition to cytoskeletal elements and the plasma membrane, the cell wall and turgor pressure equip plant and hyphal cells with structures analogous to an exoskeleton and a hydroskeleton, respectively. To quantify the physical properties of plant and hyphal cells, researchers have developed a plethora of experimental methods. This review provides an overview of experimental approaches that have been used to measure turgor pressure and to determine the mechanical properties of the plant cell wall at the subcellular level. It is completed by a glimpse into the arsenal of techniques that has been used to characterize the physical properties of cytoskeletal elements. These have mostly been used on animal cells, but we hope they will find their way into plant cell research. Finally, assays and tests to measure the generation of forces by cells and subcellular structures are discussed. PMID- 21642086 TI - Exploring the micromechanical design of plant cell walls. AB - Plants are hierarchically organized in a way that their macroscopic properties emerge from their micro- and nanostructural level. Hence, micromechanical investigations, which focus on the mechanical design of plant cell walls, are well suited for elucidating the details of the relationship between plant form and function. However, due to the complex nature of primary and secondary cell walls, micromechanical tests on the entire structure cannot provide exact values for polymer properties but must be targeted at the general mechanisms of cell wall deformation and polymer interaction. The success of micromechanical examinations depends on well-considered specimen selection and/or sample pretreatment as well as appropriate experimental setups. Making use of structural differences by taking advantage of the natural variability in plant tissue and cell structure, adaptation strategies can be analyzed at the micro- and nanoscale. Targeted genetic and enzymatic treatments can be utilized to specifically modify individual polymers without degrading the structural integrity of the cell wall. The mechanical properties of such artificial systems reveal the functional roles of individual polymers for a better understanding of the mechanical interactions within the cell wall assembly. In terms of testing methodology, in situ methods that combine micromechanical testing with structural and chemical analyses are particularly well suited for the study of the basic structure-property relationships in plant design. The micromechanical approaches reviewed here are not exhaustive, but they do provide a reasonably comprehensive overview of the methodology with which the general mechanisms underlying the functionality of plant micro- and nanostructure can be explored without destroying the entire cell wall. PMID- 21642087 TI - Effects of structural variation in xyloglucan polymers on interactions with bacterial cellulose. AB - A cellulose/xyloglucan framework is considered to form the basis for the mechanical properties of primary plant cell walls and hence to have a major influence on the biomechanical properties of growing, fleshy plant tissues. In this study, structural variants of xyloglucan have been investigated as components of composites with bacterial cellulose as a simplified model for the cellulose/xyloglucan framework of primary plant cell walls. Evidence for molecular binding to cellulose with perturbation of cellulose crystallinity was found for all xyloglucan types. High molecular mass samples gave homogeneous centimeter-scale composites with extensive cross-linking of cellulose with xyloglucan. Lower molecular mass xyloglucans gave heterogeneous composites having a range of microscopic structures with little, if any, cross-linking. Xyloglucans with reduced levels of galactose substitution had evidence of self-association, competitive with cellulose binding. At comparable molecular mass, fucose substitution resulted in a modest promotion of microscopic features characteristic of primary cell walls. Taken together, the data are evidence that galactose substitution of the xyloglucan core structure is a major determinant of cellulose composite formation and properties, with additional fucose substitution acting as a secondary modulator. These conclusions are consistent with reported structural and mechanical properties of Arabidopsis mutants lacking specific fucose and/or galactose residues. PMID- 21642088 TI - Biomechanics of plant growth. AB - Growth of turgid cells, defined as an irreversible increase in cell volume and surface area, can be regarded as a physical process governed by the mechanical properties of the cell wall and the osmotic properties of the protoplast. Irreversible cell expansion is produced by creating a driving force for water uptake by decreasing the turgor through stress relaxation in the cell wall. This mechano-hydraulic process thus depends on and can be controlled by the mechanical properties of the wall, which in turn are subject to modification by wall loosening and wall stiffening reactions. The biochemical mechanisms of these changes in mechanical wall properties and their regulation by internal signals (e.g., hormones) or external signals (e.g., light, drought stress) are at present incompletely understood and subject to intensive research. These signals act on walls that have the properties of composite materials in which the molecular structure and spatial organization of polymers rather than the distribution of mechanical stresses dictate the allometry of cell and organ growth and thus cell and organ shape. The significance of cell wall architecture for allometric growth can be demonstrated by disturbing the oriented deposition of wall polymers with microtubule-interfering drugs such as colchicine. Elongating organs (e.g., cylindrical stems or coleoptiles) composed of different tissues with different mechanical properties exhibit longitudinal tissue tensions resulting in the transfer of wall stress from inner to peripheral cell layers that adopt control over organ growth. For physically analyzing the growth process leading to seed germination, the same mechanical and hydraulic parameters as in normal growth are principally appropriate. However, for covering the influences of the tissues that restrain embryo expansion (seed coat, endosperm), an additional force and a water permeability term must be considered. PMID- 21642089 TI - Comparison of mechanical properties of four large, wave-exposed seaweeds. AB - Seaweeds have a simple structural design compared to most terrestrial plants. Nonetheless, some species have adapted to the severe mechanical conditions of the surf zone. The material properties of either tissue sections or the whole stipe of four wave-exposed seaweeds, Durvillaea antarctica, D. willana, Laminaria digitata, and L. hyperborea, were tested in tension, bending, and torsion. Durvillaea has a very low modulus of elasticity in tension (E(tension) = 3-7 MN.m(-2)) and in bending (E(bending) = 9-12 MN . m(-2)), torsion modulus (G = 0.3 MN . m(-2)) and strength (sigma(b)rk = 1-2 MN . m(-2)), combining a compliable and twistable stipe "material" with a comparatively high breaking strain (epsilon(brk) = 0.4-0.6). In comparison, the smaller stipes of Laminaria have a higher modulus of elasticity in tension (E(tension) = 6-28 MN.m(-2)) and in bending (E(bending) = 84-109 MN.m(-2)), similar strength (sigma(brk) = 1-3 MN.m( 2)), and a higher torsion modulus (G = 0.7-10 MN.m(-2)), combined with a lower breaking strain (epsilon(brk) = 0.2-0.3) than Durvillaea. Time-dependent, viscoelastic reactions were investigated with cycling tests. The tested species dissipated 42-52% of the loading energy in tension through plastic-viscoelastic processes, a finding that bears important ecological implications. Overall, there seems to be no correlation between single material properties and the size or habitat position of the tested seaweed species. PMID- 21642090 TI - Biomechanics and transgenic wood. AB - Wood, or secondary xylem, is composed mostly of three components-cellulose, hemicelluloses, and lignin. Yet this apparent simplicity is deceiving because the sophisticated arrangement of the components on various structural levels, ranging from intricate molecular architecture to defined cellular arrangements to tissue morphology, makes wood a challenging and interesting subject of biomechanical investigation. Recent advances in genetic transformation, providing easier access to wood of specifically altered composition or structure, have opened new opportunities for research on the intricate relation between material structure and composition and mechanical properties. At the same time, investigations into the mechanical properties have provided new information regarding the structural configuration of wood. The present paper reviews the work conducted in this field and outlines future perspectives and prospects for research. PMID- 21642091 TI - Wood for sound. AB - The unique mechanical and acoustical properties of wood and its aesthetic appeal still make it the material of choice for musical instruments and the interior of concert halls. Worldwide, several hundred wood species are available for making wind, string, or percussion instruments. Over generations, first by trial and error and more recently by scientific approach, the most appropriate species were found for each instrument and application. Using material property charts on which acoustic properties such as the speed of sound, the characteristic impedance, the sound radiation coefficient, and the loss coefficient are plotted against one another for woods. We analyze and explain why spruce is the preferred choice for soundboards, why tropical species are favored for xylophone bars and woodwind instruments, why violinists still prefer pernambuco over other species as a bow material, and why hornbeam and birch are used in piano actions. PMID- 21642092 TI - Fracture mechanics and its relevance to botanical structures. AB - In the field of fracture mechanics, an analytical framework has been established for understanding the mechanical failure of any structure made of inherently flawed materials. In the context of botany, this includes an extraordinarily wide variety of turgid and/or woody structures made of cellulose-based tissues, the diverse soils penetrated by their roots, and a multitude of plant-based commodities and foodstuffs. The goal of this article is to provide an overview of the theory of engineering fracture mechanics and to identify some special characteristics of wood and other plant-based materials that require further development in this area. PMID- 21642093 TI - Biomimetics and technical textiles: solving engineering problems with the help of nature's wisdom. AB - The significance of inspiration from nature for technical textiles and for fibrous composite materials is demonstrated by examples of already existing technical solutions that either parallel biology or are indeed inspired by biological models. The two different basic types of biomimetic approaches are briefly presented and discussed for the "technical plant stem." The technical plant stem is a biomimetic product inspired by a variety of structural and functional properties found in different plants. The most important botanical templates are the stems of the giant reed (Arundo donax, Poaceae) and of the Dutch rush (Equisetum hyemale, Equisetaceae). After analysis of the structural and mechanical properties of these plants, the physical principles have been deduced and abstracted and finally transferred to technical applications. Modern computer-controlled fabrication methods for producing technical textiles and for structuring the embedding matrix of compound materials render unique possibilities for transferring the complex structures found in plants, which often are optimized on several hierarchical levels, into technical applications. This process is detailed for the technical plant stem, a biomimetic, lightweight, fibrous composite material based on technical textiles with optimized mechanical properties and a gradient structure. PMID- 21642094 TI - A unified hypothesis of mechanoperception in plants. AB - The perception of mechanical stimuli in the environment is crucial to the survival of all living organisms. Recent advances have led to the proposal of a plant-specific mechanosensory network within plant cells that is similar to the previously described network in animal systems. This sensory network is the basis for a unifying hypothesis, which may account for the perception of numerous mechanical signals including gravitropic, thigmomorphic, thigmotropic, self loading, growth strains, turgor pressure, xylem pressure potential, and sound. The current state of our knowledge of a mechanosensory network in plants is reviewed, and two mechanoreceptor models are considered: a plasmodesmata-based cytoskeleton-plasma membrane-cell wall (CPMCW) network vs. stretch-activated ion channels. Post-mechanosensory physiological responses to mechanical stresses are also reviewed, and future research directions in the area of mechanoperception and response are recommended. PMID- 21642095 TI - Posture control and skeletal mechanical acclimation in terrestrial plants: implications for mechanical modeling of plant architecture. AB - Self-supporting plant stems are slender, erect structures that remain standing while growing in highly variable mechanical environments. Such ability is not merely related to an adapted mechanical design in terms of material-specific stiffness and stem tapering. As many terrestrial standing animals do, plant stems regulate posture through active and coordinated control of motor systems and acclimate their skeletal growth to prevailing loads. This analogy probably results from mechanical challenges on standing organisms in an aerial environment with low buoyancy and high turbulence. But the continuous growth of plants submits them to a greater challenge. In response to these challenges, land plants implemented mixed skeletal and motor functions in the same anatomical elements. There are two types of kinematic design: (1) plants with localized active movement (arthrophytes) and (2) plants with continuously distributed active movements (contortionists). The control of these active supporting systems involves gravi- and mechanoperception, but little is known about their coordination at the whole plant level. This more active view of the control of plant growth and form has been insufficiently considered in the modeling of plant architecture. Progress in our understanding of plant posture and mechanical acclimation will require new biomechanical models of plant architectural development. PMID- 21642096 TI - Size and function in conifer tracheids and angiosperm vessels. AB - The wide size range of conifer tracheids and angiosperm vessels has important consequences for function. In both conduit types, bigger is better for conducting efficiency. The gain in efficiency with size is maximized by the control of conduit shape, which balances end-wall and lumen resistances. Although vessels are an order of magnitude longer than tracheids of the same diameter, they are not necessarily more efficient because they lack the low end-wall resistance of tracheids with torus-margo pits. Instead, vessels gain conducting efficiency over tracheids by achieving wider maximum diameters. End-walls contributed 56-64% to total xylem resistance in both conduit types, indicating that length limits conducting efficiency. Tracheid dimensions may be more limited by unicellularity and the need to supply strength to homoxylous wood than by the need to protect against cavitation. In contrast, the greater size of the multicellular vessel is facilitated by fibers that strengthen heteroxylous wood. Vessel dimensions may be most limited by the need to restrict intervessel pitting and cavitation by air seeding. Stressful habitats that promote narrow vessels should favor coexistence of conifers and angiosperms. The evolution of vessels in angiosperm wood may have required early angiosperms to survive a phase of mechanic and hydraulic instability. PMID- 21642097 TI - Mechanical stability of trees under static loads. AB - Wind affects the structure and functioning of a forest ecosystem continuously and may cause significant economic loss in managed forests by reducing the yield of recoverable timber, increasing the cost of unscheduled thinning and clear cuttings, and creating problems in forestry planning. Furthermore, broken and uprooted trees within the forest are subject to insect attack and may provide a suitable breeding substrate, endangering the remaining trees. Therefore, an improved understanding of the processes behind the occurrence of wind-induced damage is of interest to many forest ecologists, but may also help managers of forest resources to make appropriate management decisions related to risk management. Using fundamental physics, empirical experiments, and mechanistic model-based approaches in interaction, we can study the susceptibility of tree stands to wind damage as affected by the wind and site and tree/stand characteristics and management. Such studies are not possible based on statistical approaches alone, which are not able to define the causal links between tree parameters and susceptibility to wind damage. The aim of this paper is to review the recent work done related to tree-pulling and wind tunnel experiments and mechanistic modeling approaches to increase our understanding of the mechanical stability of trees under static loading. PMID- 21642098 TI - The effect of wind exposure on the tree aerial architecture and biomechanics of Sitka spruce (Picea sitchensis, Pinaceae). AB - This paper reports on the effect of wind loading below damaging strength on tree mechanical and physical properties. In a wind-exposed Sitka spruce stand in western Scotland, 60 trees at four different levels of wind exposure (10 m, 30 m, 50 m, 90 m from edge) were characterized for stem and crown size and shape and mechanical properties, including structural Young's modulus (E(struct)), natural frequency, and damping ratio. E(struct) increased from the stand edge to the mid forest, but with a large inter-tree variation. Swaying frequency and damping ratio of the trees also increased with distance from edge. Wind-exposed edge trees grew shorter, but more tapered with an overall lower E(struct), allowing for greater flexural stiffness at the stem base due to the larger diameter and for higher flexibility in the crown region of the stem. The trees at the middle of the stand compensated for their increased slenderness with a higher E(struct). Thus, for the different requirements for wind-firmness at stand edge and mid forest, an adapted combination of tree form and mechanical properties allows the best withstanding of wind loads. The results show the requirement to understand the different strategies of trees to adapt to environmental constraints and the heterogeneity of their growth reactions in response to these strategies. PMID- 21642099 TI - Mechanical stability of trees under dynamic loads. AB - Tree stability in windstorms and tree failure are important issues in urban areas where there can be risks of damage to people and property and in forests where wind damage causes economic loss. Current methods of managing trees, including pruning and assessment of mechanical strength, are mainly based on visual assessment or the experience of people such as trained arborists. Only limited data are available to assess tree strength and stability in winds, and most recent methods have used a static approach to estimate loads. Recent research on the measurement of dynamic wind loads and the effect on tree stability is giving a better understanding of how different trees cope with winds. Dynamic loads have been measured on trees with different canopy shapes and branch structures including a palm (Washingtonia robusta), a slender Italian cypress (Cupressus sempervirens) and trees with many branches and broad canopies including hoop pine (Araucaria cunninghamii) and two species of eucalypt (Eucalyptus grandis, E. teretecornus). Results indicate that sway is not a harmonic, but is very complex due to the dynamic interaction of branches. A new dynamic model of a tree is described, incorporating the dynamic structural properties of the trunk and branches. The branch mass contributes a dynamic damping, termed mass damping, which acts to reduce dangerous harmonic sway motion of the trunk and so minimizes loads and increases the mechanical stability of the tree. The results from 12 months of monitoring sway motion and wind loading forces are presented and discussed. PMID- 21642100 TI - The biomechanics of browsing and grazing. AB - Terrestrial plant leaves are complex structures of composite materials. Resistance to fracture is achieved by a number of mechanisms, which operate at the molecular, cell, tissue, and structural levels. Leaves of dicots have different venation patterns and cell wall volume fractions from those of grasses, and consequently, they potentially resist fracture in different ways. Animals mechanically process plants in order to rupture the cell wall in preparation for enzymic hydrolysis, for which the imperative is to maximize new surface area and/or to expedite access to cell contents, ideally by promoting elastic fracture. The two different plant types are fed on by two different groups of organisms of very different sizes, digestive physiologies, mechanical processing abilities and properties, and nutritional requirements. Small insects can feed in or on parts of leaves, while larger mammals generally have to feed on the whole leaf. The scale of feeding also differs for the two groups of herbivores, but how this interacts with the scale of the mechanical properties of the leaf is not well understood. Plant leaves are attacked at all scales and probably can only produce generalized responses to specialized herbivores. In addition, the opportunities that these different scales of interactions open for the different herbivores remain unexplored. PMID- 21642101 TI - Plant biomechanics in an ecological context. AB - Fundamental plant traits such as support, anchorage, and protection against environmental stress depend substantially on biomechanical design. The costs, subsequent trade-offs, and effects on plant performance of mechanical traits are not well understood, but it appears that many of these traits have evolved in response to abiotic and biotic mechanical forces and resource deficits. The relationships between environmental stresses and mechanical traits can be specific and direct, as in responses to strong winds, with structural reinforcement related to plant survival. Some traits such as leaf toughness might provide protection from multiple forms of stress. In both cases, the adaptive value of mechanical traits may vary between habitats, so is best considered in the context of the broader growth environment, not just of the proximate stress. Plants can also show considerable phenotypic plasticity in mechanical traits, allowing adjustment to changing environments across a range of spatial and temporal scales. However, it is not always clear whether a mechanical property is adaptive or a consequence of the physiology associated with stress. Mechanical traits do not only affect plant survival; evidence suggests they have downstream effects on ecosystem organization and functioning (e.g., diversity, trophic relationships, and productivity), but these remain poorly explored. PMID- 21642102 TI - Pruning-induced tylose development in stems of current-year shoots of Vitis vinifera (Vitaceae). AB - Tyloses form in xylem vessels in response to various environmental stimuli, but little is known of the kinetics or regulation of their development. Preliminary investigations indicated that wounds seal quickly with tyloses after pruning of grapevine shoots. In this study, tylose development was analyzed qualitatively and quantitatively at different depths and times from pruning cuts along current year shoots of grapevines at basal, middle, and apical stem regions. Tyloses developed simultaneously within a single vessel but much separated in time among vessels. Pruning caused prodigious tylosis in vessels of grape stems, extending to approximately 1 cm deep and to 7 d after wounding, but about half of the vessels did not become completely occluded. The fraction of vessels forming tyloses was greatest in basal (85%) and least in apical (50%) regions. The depth of maximum density of tyloses was 4 mm from the cut in the basal region and 2 mm from the cut in the middle and apical regions. Tylose development was faster in the basal and middle than in the apical region. The pattern of tylose development is discussed in the context of wound repair and pathogen movement in grapevines. PMID- 21642103 TI - Ontogenetic trends in the morphological features of main stem annual shoots of Pinus pinaster (Pinaceae). AB - Phase change refers to the transition between juvenile and adult vegetative phases. The study of trees throughout their entire life span requires retrospective analyses and validates the use of a chronosequence by sequencing observations at different and successive stages. The main axis growth pattern of 62 maritime pines (Pinus pinaster) selected in a chronosequence of three stands consisting of 8-, 22-, and 48-yr-old trees was analyzed retrospectively. Comparison of measured features (length, number of axillary products, reproductive organs) at common ages from the three stands supported the validity of using these data to form a continuous chronosequence. Endogenous trends in tree development are revealed free from variability due to annual growth conditions. Two main phases of development corresponding respectively to the juvenile vegetative and adult reproductive stages were identified, and the transition between both occurred in 9-yr-old trees. The relevance of these two phases and more generally the notion of phase changes are discussed in light of observed trends in the values of studied growth and branching parameters that may either show gradual variations (such as length of annual shoot) or a distinctive expression in one or the other phase (such as presence of female cones). PMID- 21642104 TI - Bordered pit structure and function determine spatial patterns of air-seeding thresholds in xylem of Douglas-fir (Pseudotsuga menziesii; Pinaceae) trees. AB - The air-seeding hypothesis predicts that xylem embolism resistance is linked directly to bordered pit functioning. We tested this prediction in trunks, roots, and branches at different vertical and radial locations in young and old trees of Pseudotsuga menziesii. Dimensions of bordered pits were measured from light and scanning electron micrographs, and physiological data were from published values. Consistent with observations, calculations showed that earlywood tracheids were more resistant to embolism than latewood tracheids, mainly from earlywood having stretchier pit membranes that can distend and cover the pit aperture. Air seeding that occurs in earlywood appears to happen through gaps between the torus edge and pit border, as shown by the similar calculated pressures required to stretch the membrane over the pit aperture and to cause embolism. Although bordered pit functioning was correlated with tracheid hydraulic diameter, pit pore size and above all pit aperture constrained conductivity the most. From roots to branches and from the trunk base to higher on the trunk, hydraulic resistance of the earlywood pit membrane increased significantly because of a decrease in the size of the pit aperture and size and number of margo pores. Moreover, overall wood conductivity decreased, in part due to lower pit conductivity and a decrease in size and frequency of pits. Structural and functional constraints leading to the trade-off of efficiency against safety of water transport were also demonstrated at the individual pit level, with a positive correlation between pit membrane resistance on an area basis and the pressure differential required to cause membrane stretching, a characteristic that is essential for pit aspiration. PMID- 21642105 TI - Sclerophylly in two contrasting tropical environments: low nutrients vs. low rainfall. AB - The defining characteristics of sclerophylly are mechanical (e.g., hardness, toughness, stiffness), but little is known about how they vary in contrasting environments and contribute to the adaptiveness of sclerophylly. Here we investigate how the degree and nature of sclerophylly in terms of leaf mechanics differ between vegetation of two contrasting stressful environments, maquis on nutrient-deficient, moist sites and dry forest on moderate-nutrient, drier sites. We measured toughness, strength, and stiffness at the level of the whole leaf ("structural") and per unit thickness ("material"). Leaves of maquis plants were on average structurally stiffer, stronger, and tougher than those in dry forest. There was little difference in material properties between habitats, and leaf thickness was the main contributor to differences in structural mechanical properties between habitats. Flexural stiffness varied most among species and habitats, correlating strongly with leaf mass per area and thickness. We suggest that having thicker leaves allows efficient packaging of biomass to reduce branching costs in sunny but stressful environments, with subsequent impacts on structural mechanical properties. Sclerophylly is probably a complex phenomenon, however, with its mechanical constitution arising from both evolved mechanical properties that confer protection or resistance to stress and nonadaptive mechanical consequences of adaptation to stressful environments. PMID- 21642106 TI - Influence of substrate type and microsite availability on the persistence of foxtail pine (Pinus balfouriana, Pinaceae) in the Klamath Mountains, California. AB - The persistence of poor competitors within species-rich assemblages is often tied to habitat heterogeneity. Here, the persistence of foxtail pine (Pinus balfouriana) in the Klamath Mountains of northern California was addressed using a two-step approach. First, the response of foxtail pine to shading from six co occurring conifers was examined using two morphological indices. Foxtail pine increased the height to the first branch that supported foliage, and this branch was shorter when compared with those on all other sampled conifers, suggesting that foxtail pine is a poor competitor for light. Second, three hypotheses to explain foxtail pine persistence were tested: habitat heterogeneity at large spatial scales (substrate hypothesis), habitat heterogeneity at small spatial scales (microsite hypothesis), and the long lifespan of foxtail pine (successional hypothesis). Habitat heterogeneity at multiple spatial scales favored the persistence of foxtail pine. At large spatial scales, ultramafic substrates affected the importance and competitive abilities of shade-tolerant conifers. At small spatial scales, species richness, species diversity (H'), and stand density were positively correlated with microsite availability. No support was found for the successional hypothesis. Results are subsequently linked with general hypotheses of species coexistence in species-rich assemblages. PMID- 21642107 TI - Low clouds and cloud immersion enhance photosynthesis in understory species of a southern Appalachian spruce-fir forest (USA). AB - High-altitude forests of the southern Appalachian Mountains (USA) are frequently immersed in clouds, as are many mountain forests. They may be particularly sensitive to predicted increases in cloud base altitude with global warming. However, few studies have addressed the impacts of immersion on incident sunlight and photosynthesis. Understory sunlight (photosynthetically active radiation, PAR) was measured during clear, low cloud, and cloud-immersed conditions at Mount Mitchell and Roan Mountain, NC (USA) along with accompanying photosynthesis in four representative understory species. Understory PAR was substantially less variable on immersed vs. clear days. Photosynthesis became light-saturated between ~100 and 400 MUmol . m(-2) . s(-1) PAR for all species measured, corresponding closely to the sunlight environment measured during immersion. Estimated daily carbon gain was 26% greater on clear days at a more open canopy site but was 22% greater on immersed/cloudy days at a more closed canopy site. F(v)/F(m) (maximum photosystem II efficiency) in Abies fraseri seedlings exposed to 2.5 min full sunlight was significantly reduced (10%), indicating potential reductions in photosynthesis on clear days. In addition, photosynthesis in microsites with canopy cover was nearly 3-fold greater under immersed (2.6 mmol . m(-2) . h(-1)) vs. clear conditions (0.9 mmol . m(-2) . h(-1)). Thus, cloud immersion provided more constant PAR regimes that enhanced photosynthesis, especially in shaded microsites. Future studies are needed to predict the survival of these refugial forests under potential changes in cloud regimes. PMID- 21642108 TI - On the developmental dependence of leaf respiration: responses to short- and long term changes in growth temperature. AB - Using measurements of leaf respiratory O(2) uptake (R), we investigated whether immature and mature Arabidopsis thaliana (ecotype Columbia) leaves differed in their response to temperature. Confocal microscopy (using plants with mitochondrially targeted green fluorescent protein [GFP]) was used to determine whether ontogenetic changes in R are associated with concomitant changes in mitochondrial morphology/abundance. Comparisons were made of warm-grown (25/20 degrees C) leaves, warm-grown leaves shifted to cold (5 degrees C) for 10 days, and cold-developed leaves. Short-term Q(10) values and the ability to cold acclimate were determined. In warm-grown plants, rates of R per mass were highest in immature leaves, decreasing as leaves developed. Moreover, although mitochondrial size (5.6-6.5 MUm(3)) remained constant during development, mitochondrial number per MUm(3) declined from 0.01 to 0.003 as leaves expanded (i.e., mitochondrial density decreased). Immature and mature leaves did not differ in Q(10) values but did differ in their ability to cold-acclimate. Whereas mature leaves had clear evidence of cold acclimation (e.g., when measured at 25 degrees C, R was highest in cold-developed leaves), young leaves had none. Collectively, the results highlight the changes in rates of R, mitochondrial density, and biomass allocation associated with leaf development and that changes in respiratory flux associated with acclimation only take place within mature tissues. PMID- 21642109 TI - Genetic structure of experimental populations and reproductive fitness in a heterocarpic plant Atriplex tatarica (Chenopodiaceae). AB - Atriplex tatarica is a heterocarpic species of disturbed habitats. Seeds of Atriplex tatarica do not germinate immediately after shedding, but may remain in a dormant but viable state indefinitely. We investigated whether there were genetic and fitness differences between plants derived from seeds of the different fruit types germinated in different temperatures and salinities. Seeds that germinated in optimal and suboptimal conditions differed significantly in their genetic composition due, in part, to their source population. Seeds that germinated in the suboptimal conditions produced more homozygous plants. Plants that were primarily heterozygous were generated from nondormant fruit types as well as from fruits that germinated in the optimal conditions. Moreover, there was a positive correlation between the degree of heterozygosity and plant fitness measured as the mass of the stem and reproductive structures. In conclusion, the genetic variation of natural populations may be at least partly due to the ability of particular seed genotypes to germinate in the specific environmental conditions of a particular locality. In some circumstances, the process of differential germination may select not only for genetic variability but also for higher fitness if heterozygosity-fitness correlations are present. PMID- 21642110 TI - Real-time patterns of pollen flow in the wild-service tree, Sorbus torminalis (Rosaceae). III. Mating patterns and the ecological maternal neighborhood. AB - Understanding the role of mother plants as pollen recipients in shaping mating patterns is essential for understanding the evolution of populations and in particular to predict the consequence of habitat fragmentation. Here, we investigated variation in mating patterns due to maternal phenotypic traits, phenological variance, and landscape features in Sorbus torminalis, a hermaphroditic, insect-pollinated and low-density, European temperate forest tree. The diversity and composition of pollen clouds received by maternal trees in S. torminalis were mainly determined by their conspecific neighborhood: isolated individuals sample more diversity through more even paternal contributions, low relatedness among paternal genes, and high rates of long distance pollen dispersal within their progenies. Maternal phenotypic traits related to pollinator attractiveness also had an effect, but only when competition was strong: in this case, larger mother trees with more flowers sampled more diversity. The floral architecture of S. torminalis, with multiple seeded fruit, strongly shaped mating patterns, with higher levels of correlated paternity among seeds belonging to the same fruit (30% full sibs) than among seeds belonging to different fruits (14% full sibs). Finally, flowering phenology affected the distribution of diversity among maternal pollen clouds, but the earliest and latest mother trees did not receive less diversity of pollen than the others. PMID- 21642111 TI - Limits to reproductive success of Sarracenia purpurea (Sarraceniaceae). AB - Plant biologists have an enduring interest in assessing components of plant fitness and determining limits to seed set. Consequently, the relative contributions of resource and pollinator availability have been documented for a large number of plant species. We experimentally examined the roles of resource and pollen availability on seed set by the northern pitcher plant Sarracenia purpurea. We were able to distinguish the relative contributions of carbon (photosynthate) and mineral nutrients (nitrogen) to reproductive success. We also determined potential pollinators of this species. The bees Bombus affinis and Augochlorella aurata and the fly Fletcherimyia fletcheri were the only floral visitors to S. purpurea that collected pollen. Supplemental pollination increased seed set by <10%, a much lower percentage than would be expected, given data from noncarnivorous, animal-pollinated taxa. Seed set was reduced by 14% in plants that could not capture prey and by another 23% in plants whose pitcher-shaped leaves were also prevented from photosynthesizing. We conclude that resources are more important than pollen availability in determining seed set by this pitcher plant and that reproductive output may be another "cost" of the carnivorous habit. PMID- 21642112 TI - Photographs and herbarium specimens as tools to document phenological changes in response to global warming. AB - Global warming is affecting natural systems across the world. Of the biological responses to warming, changes in the timing of phenological events such as flowering are among the most sensitive. Despite the recognized importance of phenological changes, the limited number of long-term records of phenological events has restricted research on the topic in most areas of the world. In a previous study in Boston (American Journal of Botany 91: 1260-1264), we used herbarium specimens and one season of field observations to show that plants flowered earlier as the climate warmed over the past 100 yr. In our new study, we found that two extra years of data did not strengthen the explanatory power of the analysis. Analysis of herbarium specimens without any field data yielded results similar to analyses that included field observations. In addition, we found that photographs of cultivated and wild plants in Massachusetts, data similar to that contained in herbarium specimens, show changes in flowering times that closely match independent data on the same species in the same locations. Dated photographs of plants in flower represent a new resource to extend the range of species and localities addressed in global-warming research. PMID- 21642113 TI - Effectiveness of bats as pollinators of Stenocereus stellatus (Cactaceae) in wild, managed in situ, and cultivated populations in La Mixteca Baja, central Mexico. AB - Stenocereus stellatus is an endemic, self-incompatible, columnar cactus found in central Mexico where many of its wild populations have been fragmented. As an economically important species of fruit-producing cactus, S. stellatus occurs in wild, managed in situ, and cultivated populations. The objectives of this study were to determine the effective pollinators of S. stellatus, to compare pollinator visits and reproductive parameters among the three types of populations, and to determine if nectar feeding-bats are moving among populations. Effective pollinators were the nectarivorous bats Choeronycteris mexicana, Leptonycteris curasoae, and L. nivalis. Fewer total visits per flower per night and fewer visits by Choeronycteris were observed in cultivated populations, while the opposite pattern was observed for Leptonycteris. One aggressive interaction was filmed in which Choeronycteris was physically displaced by Leptonycteris, and Choeronycteris visits were significantly affected by Leptonycteris visits. Cultivated populations received more pollen grains and had more fruit set. Variation in pollinator visits between different populations and the consequent effects on reproductive success were likely a result of competition between bat species, and differences in foraging and in sensitivity of bat species to human populations. Three marked L. curasoae traveled 15 km from their roosting site to their foraging area, and one visited cultivated and managed populations, suggesting that this species may be particularly important in moving pollen among populations. PMID- 21642114 TI - Phylogeny and evolution of Burmanniaceae (Dioscoreales) based on nuclear and mitochondrial data. AB - The mycoheterotrophic Burmanniaceae are one of the three families currently recognized in the order Dioscoreales. Phylogenetic inference using nucleotide sequences of the nuclear 18S rDNA region and the mitochondrial nad1 b-c intron revealed two well-supported, major lineages within the family, corresponding to the two tribes recognized in the family: Burmannieae and Thismieae. All data supported a strong relationship between Thismieae and Tacca (Dioscoreaceae) making both Burmanniaceae and Dioscoreaceae polyphyletic. The three largest Burmanniaceae genera, Burmannia, Gymnosiphon, and Thismia, are paraphyletic. The splitting of Burmanniaceae into Burmannieae and Thismieae indicates two independent origins of mycoheterotrophy and correlated loss of chlorophyll in Dioscoreales. In the genus Burmannia, in which many species still contain chlorophyll, the achlorophyllous species are nested in between the autotrophic species, suggesting many independent changes from autotrophy to heterotrophy or vice versa. A Bayesian relative rates test on the 18S rDNA data showed considerable variation in substitution rates among Burmanniaceae. The substitution rates in all Thismieae and many Burmannieae are significantly faster than in Dioscoreaceae, but there seems to be no correlation between rate increases and the loss of photosynthesis. PMID- 21642115 TI - Phylogeny, taxonomic affinities, and biogeography of Penstemon (Plantaginaceae) based on ITS and cpDNA sequence data. AB - The large and diverse genus Penstemon (ca. 271 species) is endemic to North America and has been divided into six subgenera primarily based on anther dehiscence patterns. Species of Penstemon are known to be pollinated by a variety of insects (hymenopterans, lepidopterans, dipterans) and hummingbirds. Nucleotide sequence data from ITS and two noncoding regions of chloroplast DNA were used to reconstruct the phylogeny of Penstemon. Trees generated from nuclear and chloroplast DNA sequences are incongruent, which is probably the result of hybridization, and not fully resolved, which is likely due to a rapid evolutionary radiation. Penstemon represents a recent continental radiation where speciation has resulted primarily from evolutionary adaptations to ecological niches such as pollinator specialization. The results from these analyses show that the current circumscription of subgenera and sections needs revision to reflect more closely the evolutionary relationships of species. Specifically, species in subgenera Saccanthera, Habroanthus, and Penstemon are polyphyletic. These results also confirm the independent origin of hummingbird floral morphology in 10 clades. PMID- 21642116 TI - Intrageneric phylogeny of Capsella (Brassicaceae) and the origin of the tetraploid C. bursa-pastoris based on chloroplast and nuclear DNA sequences. AB - Polyploidization, often accompanied by hybridization, has been of major importance in flowering plant evolution. Here we investigate the importance of these processes for the evolution of the tetraploid crucifer Capsella bursa pastoris using DNA sequences from two chloroplast loci as well as from three nuclear low-copy genes. The near-absence of variation at the C. bursa-pastoris chloroplast markers suggests a single and recent origin of the tetraploid. However, despite supporting a single phylogeny, chloroplast data indicate that neither of the extant Capsella diploids is the maternal parent of the tetraploid. Combined with data from the three nuclear loci, our results do not lend support to previous hypotheses on the origin of C. bursa-pastoris as an allopolyploid between the diploids C. grandiflora and C. rubella or an autopolyploid of C. grandiflora. Nevertheless, for each locus, some of the C. bursa-pastoris accessions harbored C. rubella alleles, indicating that C. rubella contributed to the gene pool of C. bursa-pastoris, either through allopolyploid speciation or, more likely, through hybridization and introgression. To our knowledge, this study is the first of a wild, nonmodel plant genus that uses a combination of chloroplast and multiple low-copy nuclear loci for phylogenetic inference of polyploid evolution. PMID- 21642117 TI - Role of accelerated style senescence in pathogen defense. AB - Plants, like animals, suffer from a variety of diseases that are transmitted via their sexual organs. In many species, the flowers senesce rapidly after pollination or fertilization. In ongoing studies of the impacts of a transposon insertional mutation in the gene that encodes the most abundant isoform of a major group-1 pollen allergen of maize, we found that pollen tubes with the mutant allele grow significantly slower in vivo than pollen with the wild-type allele. Here, we report that under field conditions, maize silks (styles) pollinated with pollen bearing the slower-growing mutant allele take significantly longer to senesce, and the resulting ears (infructescences) have dramatically higher incidence of "fungal ear rot" disease than silks pollinated with pollen bearing the wild-type allele. Because ear rot fungi gain access to the developing ear by growing on and through the silks, we propose that accelerated senescence of silks after fertilization is a defense against pathogens such as those causing ear rot. In addition, we divided the silks on each ear into two halves and experimentally varied the type of pollen (wild type, mutant, unpollinated) that was placed onto each half of the silks. Senescence of unpollinated silks was accelerated when ovaries on the other half of the ear were fertilized. PMID- 21642118 TI - Oil bodies in leaf mesophyll cells of angiosperms: overview and a selected survey. AB - Neutral (storage) oil bodies occur in leaf mesophyll cells of many angiosperms, but their literature has been largely forgotten. We review this literature and provide a survey of 302 species and hybrids from mostly north-central US species representing 113 families. Freehand cross sections of fresh leaves stained with Sudan IV verified the presence of oil. In 71 species from 24 families we observed 1-15 oil bodies per mesophyll cell. The eudicot families Asteraceae, Caprifoliaceae, Lamiaceae, and Rosaceae had the highest number of species with oil bodies, whereas few or no species in the Apiaceae, Betulaceae, Fabaceae, and Scrophulariaceae had them. Only three of 19 monocot species sampled had oil bodies. Repeat sampling of a Malus (crabapple) cultivar and a Euonymus species showed conspicuous oil bodies in mid-summer and also in mid-autumn in both attached and recently shed leaves. Oil bodies in leaf mesophyll cells are conspicuous (visible in hand cross sections using moderate magnification in unstained water mounts) in numerous species, and they occur throughout the growing season in at least some species. Neutral oil bodies in leaf mesophyll cells are not mentioned in contemporary textbooks and advanced works, but they deserve recognition as significant cellular components of many taxa, in which they may be significant sources of commercial oils. PMID- 21642119 TI - Medial pith cells per meter in twigs as a proxy for mitotic growth rate ({Phi}/m) in the apical meristem. AB - The Phi model for plant mating system evolution proposes a causal link between Phi, the number of mitoses that occur within a plant's lifetime from zygote to gamete production, and constraints on the evolution of inbreeding depression and thereby on the evolution of plant mating systems. Through its use of plant stature, the Phi model emphasizes the important role of morphology in creating developmental and genetic constraints on plant evolution. However, the estimation of Phi itself is likely to be extraordinarily complex. Here I describe a protocol for estimating Phi per meter linear growth by an apical meristem (Phi/m) using medial pith cells from mature internodes of twigs. While such cells are produced by the apical meristem, during internode elongation, these pith cells also undergo further mitoses, and thus their measurement can only approximate a "true" Phi/m via the application of a multiplier (the adjustment ratio) that partially corrects for the occurrence of cell divisions and cell growth beyond the apical meristem. I applied this method to Delonix regia (Caesalpiniaceae) and derived several adjustment ratios from the literature. Because variation in Phi/m can have profound evolutionary implications, I also examined interspecific and intraspecific variation as well as within-individual variation in Phi/m. Conifers apparently have lower Phi/m than do angiosperms, while 20% of the total variance in Phi/m for D. regia was found among individual trees, with the remainder found within trees. Given the large differences in stature between "high-Phi" plants such as trees and "low-Phi" plants such as herbs, these results support the idea that the total per-generation mutation rate for high-Phi plants is likely to be many times higher than that for low-Phi plants. PMID- 21642120 TI - Delayed fertilization and pollen-tube growth in pistils of Fagus japonica (Fagaceae). AB - In contrast to most angiosperms, in which fertilization occurs 1 or 2 days after pollination, in some plant orders, including the Fagales, fertilization is delayed from 4 days to more than 1 year, raising questions regarding why fertilization is delayed and where and how pollen tubes remain in the pistil during the delay. To answer these questions, we investigated pollen-tube growth in pistils of Fagus japonica (Fagaceae), which are tricarpellate and have six ovules, using light, fluorescence, and scanning electron microscopy. The ovules were immature at the time of pollination and required 5 weeks to become fully developed. During this 5 weeks, pollen tubes grew from the stigma to the embryo sac in association with the development of ovules and intermittently in three steps with two growth-cessation sites, i.e., on the funicle and near the micropyle. The number of pollen tubes was gradually reduced from many to one at the two growth-cessation sites, and fertilization occurred in one ovule that apparently developed earlier than the others in the pistil. Thus, delayed fertilization plays an important role in gametophyte competition and selection leading to nonrandom fertilization. Intermittent pollen-tube growth is also likely widespread in angiosperms because it is known in other Fagales and an unrelated order Garryales. PMID- 21642121 TI - GIS-based characterization of the geographic distributions of wild and cultivated populations of the Mesoamerican fruit tree Spondias purpurea (Anacardiaceae). AB - Humans are having a profound impact on the geographic distributions of plant populations. In crop species, domestication has been accompanied by the geographic expansion of cultivated populations relative to their wild ancestors. We used a geographical information system (GIS)-based approach to investigate differences in the environmental factors characterizing the geographic distributions of cultivated and wild populations of the Mesoamerican fruit tree Spondias purpurea. Locality data for 86 cultivated and 28 wild S. purpurea populations were used in conjunction with environmental data layers and Maxent, a maximum entropy application for predicting species distributions. Interpredictivity analyses and principal components analysis revealed that the predicted distribution of wild S. purpurea is nested within the cultivated distribution and that the ecological niche (defined by environmental characteristics) of cultivated S. purpurea has expanded relative to that of wild populations. Significant differences between wild and cultivated populations were detected for five environmental variables, corresponding to the expansion of S. purpurea during the domestication process from its native habitat in the Mesoamerican tropical dry forests into less seasonal habitats. These data suggest that humans have altered the range of habitats occupied by cultivated S. purpurea populations relative to their wild progenitors. PMID- 21642122 TI - Inbreeding effects on blossom volatiles in Cucurbita pepo subsp. texana (Cucurbitaceae). AB - Self-pollination by plants gives rise to inbreeding depression. There is increasing recognition that plant inbreeding can have significant implications for interactions between plants and other organisms, including insects and pathogens. Many of these interactions are mediated by plant-derived volatiles, but the effects of inbreeding on volatile production have not previously been investigated. We examined variation in flower volatile production by the wild gourd Cucurbita pepo subsp. texana as a function of inbreeding, sex of the flower, and maternal line. We compared first-generation selfed progeny to outcrossed progeny to assess variation in blossom volatiles due to mating system. Our data indicate that self-pollination reduces total volatile production and changes the relative composition of individual compounds released by C. pepo subsp. texana blossoms. These findings have potentially important implications for interactions between C. pepo subsp. texana and its pollinators and herbivores including diabroticite cucumber beetles, which vector the bacterial pathogen Erwinia tracheiphila-because previous studies have shown that a number of the individual compounds that vary with inbreeding level can influence insect behavior. We also found significant differences between the volatile profiles of male and female flowers and across maternal families. PMID- 21642123 TI - Ecological correlates of secondary sexual dimorphism in Salix glauca (Salicaceae). AB - The ecological causation hypothesis for secondary sexual dimorphism was tested in Salix glauca, a dioecious willow shrub. Plants growing in a Colorado Rocky Mountain (USA) krummholz mosaic of mesic and xeric patches were monitored for four consecutive years. Ecological causation is predicated on unique resource demands associated with sexual function. In S. glauca, seeds have twofold higher N and P concentrations compared to pollen. P, but not N, allocation costs differed between sexes at plant and flower scales. Ecological causation also predicts spatial segregation of sexes along underlying habitat gradients. In five populations of S. glauca, sexes displayed significant spatial segregation. The theory also predicts that sexes differ in performance across gradients of environmental stress or resource availability. Consistent with this hypothesis, females had lower drought tolerance than males under years of extreme aridity. Furthermore, over 10 years, annual shoot growth for females was greatest in mesic habitat patches, while males grew at a consistent rate regardless of habitat aridity. Because current shoot growth is correlated with future catkin production, habitat specialization likely provides a fitness payoff in females. Overall, this long-term study provides some of the strongest evidence to date for ecological causation of secondary sexual dimorphism in plants. PMID- 21642124 TI - Salt tolerance and osmotic adjustment of Spartina alterniflora (Poaceae) and the invasive M haplotype of Phragmites australis (Poaceae) along a salinity gradient. AB - An invasive variety of Phragmites australis (Poaceae, common reed), the M haplotype, has been implicated in the spread of this species into North American salt marshes that are normally dominated by the salt marsh grass Spartina alterniflora (Poaceae, smooth cordgrass). In some European marshes, on the other hand, Spartina spp. derived from S. alterniflora have spread into brackish P. australis marshes. In both cases, the non-native grass is thought to degrade the habitat value of the marsh for wildlife, and it is important to understand the physiological processes that lead to these species replacements. We compared the growth, salt tolerance, and osmotic adjustment of M haplotype P. australis and S. alterniflora along a salinity gradient in greenhouse experiments. Spartina alterniflora produced new biomass up to 0.6 M NaCl, whereas P. australis did not grow well above 0.2 M NaCl. The greater salt tolerance of S. alterniflora compared with P. australis was due to its ability to use Na(+) for osmotic adjustment in the shoots. On the other hand, at low salinities P. australis produced more shoots per gram of rhizome tissue than did S. alterniflora. This study illustrates how ecophysiological differences can shift the competitive advantage from one species to another along a stress gradient. Phragmites australis is spreading into North American coastal marshes that are experiencing reduced salinities, while Spartina spp. are spreading into northern European brackish marshes that are experiencing increased salinities as land use patterns change on the two continents. PMID- 21642125 TI - Are differences in breeding mechanisms and fertility among populations contributing to rarity in Grevillea rhizomatosa (Proteaceae)? AB - Plant breeding systems are seldom studied across the breadth of a species' range. For many systems, this precludes an informed assessment of the evolutionary biology of a species, particularly of the factors that shape fecundity. Grevillea rhizomatosa is a threatened species of shrub known only from a 7 * 8 km area and c. 2000 plants in northern New South Wales, Australia. The species reproduces asexually from rhizomatous suckers, and fruit are only produced in a few populations. Over two flowering seasons, we investigated the extent of sexual reproduction and the mechanisms of infertility in five populations that span the range of the species. Seed were produced in three of the five populations. The breeding system varied among populations from obligate outcrossing to facultative outcrossing to fully sterile. Fruit to flower ratios were below 0.13 in the fertile populations but within the range found in other species of Grevillea. Pollinator limitation was not linked to infertility. Stigmatic opening and receptivity were functional in all populations. Interpopulation crosses using fertile pollen failed to recover fertility in an infertile population. A breakdown in female and male fertility mechanisms was found including a fault in the mechanical release of pollen from anthers, <10% viable pollen, and a post pollen-deposition event that prevents fertile pollen from effecting seed-set. In the infertile populations pollen was not released from anthers, resulting in flowers projecting barren pollen-presenters. Sexually reproducing populations are threatened by the incursion of asexual forms that may be favored by frequent disturbance from wild-fires. PMID- 21642126 TI - Nectar reward and advertisement in hummingbird-pollinated Silene virginica (Caryophyllaceae). AB - We tested for an association between nectar and various floral traits and investigated their roles as primary and secondary pollinator attractants in hummingbird-pollinated Silene virginica. Our goal was to gain insight into the mechanisms of pollinator-mediated selection that underlies floral trait divergence within the genus. In a field population of S. virginica, we measured five floral and eight vegetative traits and quantified nectar volume, nectar sugar concentration, and total sugar reward (nectar volume * nectar sugar concentration). All three components of nectar reward were positively correlated to flower size, and nectar volume varied significantly among individuals within the population. To ascertain whether the correlation of specific floral traits with nectar reward influences the behavior of the primary pollinator of S. virginica, the ruby-throated hummingbird, Archilochus colubris, we investigated whether A. colubris preferred the expression of floral traits associated with high nectar volume and total sugar reward. We accomplished this by constructing floral arrays consisting of artificial flowers that had equal nectar quantity and total sugar reward but that differed in petal area and corolla tube diameter, which were positively correlated with nectar quantity and total sugar reward in our field study. In observations of visitation frequencies to the various floral trait combinations, hummingbirds preferentially visited artificial floral phenotypes with larger petal displays, with the greatest preference for floral phenotypes with both larger petals and wider corolla-tube diameters. This association between primary and secondary floral attractants and hummingbird discrimination of floral features supports the concept that the floral traits of S. virginica reflect pollinator-mediated selection by the principal pollinator. PMID- 21642127 TI - Generic delimitation and relationships in Ebenaceae sensu lato: evidence from six plastid DNA regions. AB - Phylogenetic relationships of the pantropical family Ebenaceae s.l. were investigated using plastid DNA sequence data from six regions: atpB, matK, ndhF, trnK intron, trnL intron, and trnL-trnF spacer. Sampling included representatives of all currently recognized genera of Ebenaceae, Diospyros, Euclea, and Lissocarpa, and nearly all taxa that were previously recognized at the generic level, e.g., Cargillia, Gunisanthus, Maba, Macreightia, Royena, and Tetraclis. Our results strongly support monophyly of Ebenaceae s.l. and demonstrate that the previous infrafamilar classifications of the family do not circumscribe monophyletic groups. A new infrafamilial classification based on a phylogenetic approach is proposed here and consists of two subfamilies, Lissocarpoideae and Ebenoideae, and four genera, Lissocarpa, Euclea, Royena, and Diospyros. Relationships and potential synapomorphic characters are discussed and summarized. This study supports a western Gondwanan origin of family and indicates that both vicariant and long-distance dispersal events played an important role in attaining current distributions. PMID- 21642128 TI - A molecular phylogenetic study of southern African Apiaceae. AB - It has been suggested that southern Africa is the origin of the predominantly herbaceous Apiaceae subfamily Apioideae and that the woody habit is plesiomorphic. We expand previous molecular phylogenetic analyses of the family by considering all but three of the approximately 38 genera native to southern Africa, including all genera whose members, save one, have a woody habit. Representatives of five other genera are included because they may be closely related to these southern African taxa. Chloroplast DNA rps16 intron and/or nuclear rDNA ITS sequences for 154 accessions are analyzed using maximum parsimony, Bayesian, and maximum likelihood methods. Within Apioideae, two major clades hitherto unrecognized in the subfamily are inferred. The monogeneric Lichtensteinia clade is sister group to all other members of the subfamily, whereas the Annesorhiza clade (Annesorhiza, Chamarea, and Itasina) plus Molopospermum (and Astydamia in the ITS trees) are the successive sister group to all Apioideae except Lichtensteinia. Tribe Heteromorpheae is expanded to include Pseudocarum, "Oreofraga" ined., and five genera endemic to Madagascar. The southern African origin of subfamily Apioideae is corroborated (with subsequent migration northward into Eurasia along two dispersal routes), and the positions of the herbaceous Lichtensteinia and Annesorhiza clades within the subfamily suggest, surprisingly, that its ancestor was herbaceous, not woody. PMID- 21642129 TI - Utility of high resolution x-ray computed tomography (HRXCT) for paleobotanical studies: an example using London Clay fruits and seeds. AB - High resolution x-ray computed tomography (HRXCT) was used to image pyritized fossil fruits from the Lower Eocene London Clay flora to test the utility of this technique for paleobotanical application. The combination of carbon-pyrite preservation and void spaces between fruit and seed layers within fossils provides differences in density and composition that enable excellent imaging. Fossil fruits of Palaeorhodomyrtus subangulata (Bowerbank) Reid & Chandler (Myrtaceae) were investigated in situ within their silicone fluid conservation medium, which protects these unstable fossils from oxygen and humidity. HRXCT recovers taxonomically informative anatomical and morphological detail and provides a means of nondestructive examination of delicate type materials and other important specimens. These results suggest that HRXCT will be applicable to a broad spectrum of pyritized fossils to record structural details in inherently unstable materials. PMID- 21642130 TI - Chloroplast DNA phylogeography of Photinia glabra (Rosaceae) in Japan. AB - Climate changes during glacial periods have had significant effects on the present geographic distribution of plant species. To elucidate the evolutionary history of a plant species with a disjunctive distribution, we investigated the geographic distribution patterns of cpDNA haplotypes in Photinia glabra (Rosaceae) growing in broadleaved evergreen forests in Japan. We examined cpDNA in 42 populations of P. glabra sampled over a geographic range that included Kinki and its surrounding areas and the disjunctive regions in the Amakusa Islands. Both areas had unique cpDNA haplotypes. Moreover, the AMOVA revealed that a large proportion of the total variance (51%, P < 0.001) could be explained by differences among these regions. These results suggest a past fragmentation of this plant species into two separate refugia: southwestern Kyushu and Kinki, including the surrounding area, during the Quaternary glacial periods. A particularly interesting result was that in the southern disjunct distribution in the Amakusa Islands, the genetic subdivision (Phi(CT) = 1.00, P < 0.001) appears to lie between the populations from nearly contiguous islands located across a fairway only approximately 80 to 150 m in width. PMID- 21642131 TI - Specialized structures in the leaf epidermis of basal angiosperms: morphology, distribution, and homology. AB - The morphology of specialized structures in the leaf epidermis of 32 species of basal (ANITA: Amborella, Nymphaeales, Illiciales, Trimeniaceae, and Austrobaileyaceae) angiosperms, representing all seven families and 11 of 14 genera, was investigated using light and scanning electron microscopy. Distribution, density, and size of structures were also measured, and character evolution was analyzed. Hydropotes are a synapomorphy of Nymphaeales and ethereal oil cells are a synapomorphy of Austrobaileyales, but uniseriate nonglandular trichomes appear to have arisen independently several times. Specialized structures are frequently characterized by adjacent epidermal cells that have striking similarities in their form and arrangement (i.e., architecture) to subsidiary cells of certain types of stomatal complexes. Additionally, forms intermediate to oil cells and stomata, to trichomes and stomata, and to hydropotes and oil cells are present in some taxa. Thus, all of these specialized structures and their adjacent epidermal cells form complexes that may be homologous with, and evolutionarily derived from stomatal complexes, and the specialized structure, or portion thereof, may be homologous to the stoma or guard mother cell. Improved knowledge of the morphology and evolution of these structures in the earliest branching extant angiosperm lineages has a bearing on many diverse areas of botany. PMID- 21642132 TI - Comparative anatomy of root meristem and root cap in some species of Podostemaceae and the evolution of root dorsiventrality. AB - In the unusual aquatic Podostemaceae, the root is the leading organ of the plant body and is variously compressed and submerged as it adheres to rock surfaces in rapid water. In an anatomical comparison of the root apical meristems and root caps of 33 species that represent the major lineages of the family, the dorsiventrality of root meristems varied and was classified into four patterns: (1) The root cap is produced outward from a nearly radially symmetrical meristem. (2) The meristem and root cap are markedly dorsiventral; the outermost cells of the hood-shaped cap are acroscopic derivatives from bifacial initials on the ventral side, while the pattern on the dorsal side is similar to pattern 1. (3) Bifacial initials are on both the dorsal and ventral sides. (4) No root cap is present. An evolutionary polarity may be evident from pattern 1 to 2 and then to 3. Pattern 2 arose in the early evolution of the subfamily Podostemoideae and subsequently, pattern 3 arose in species with crustose roots, while the least specialized pattern 1 is retained in Tristichoideae and Weddellinoideae. Pattern 4 characterized by caplessness may have appeared recurrently in Tristichoideae and Podostemoideae. These evolutionary changes in the meristem preceded the specialization of external root morphologies. PMID- 21642133 TI - Structure and development of Nostoc strands in Leiosporoceros dussii (Anthocerotophyta): a novel symbiosis in land plants. AB - The presence of Nostoc in longitudinally oriented schizogenous canals is a feature that separates Leiosporoceros from all other hornworts and represents a novel symbiotic arrangement in land plants. In surface view, Nostoc canals are visible as elongated, dichotomously branched blue-green strands. All other hornworts develop numerous discrete globose colonies through continuous production of mucilage clefts as avenues for multiple invasions within a single thallus. To elucidate the anatomy and development of the unusual Nostoc strands in Leiosporoceros, we examined sporeling development in culture and the structure of strands in field-collected plants using light and electron microscopy. Rosette like sporelings have mucilage clefts scattered along swollen apices. All field specimens were strap-shaped, contained Nostoc, and lacked mucilage clefts. Nostoc strands are located in the center of the thallus and develop behind the apical cell by separation of the middle lamella between apical derivatives. Strands elongate and branch in synchrony with apical growth, and thus only a single invasion is required for strand production. Two distinct ultrastructural morphotypes in the collections suggest nonspecificity of Nostoc. We speculate that Nostoc enters the thallus in the sporeling stage through mucilage clefts, and once colonies are established, cleft production ceases. PMID- 21642134 TI - An architectural model for Eleocharis: morphology and development of Eleocharis cellulosa (Cyperaceae). AB - Species of Eleocharis are prominent in aquatic and wetland habitats and serve as models for study of physiological adaptations to aquatic environments. The genus has an unusual morphology because the major photosynthetic organ is the stem. In order to define an architectural model for the genus to understand the evolution of this morphology, we examined mature morphology and development of E. cellulosa in living and fixed material using light and scanning electron microscopy. Eleocharis cellulosa has sympodial, vertical shoots that produce the photosynthetic culms and horizontal shoots that mix monopodial and sympodial development. Each sympodial unit produces three bracts, an elongated photosynthetic internode, then a fourth bract and an inflorescence that either aborts on vegetative culms or expands on reproductive culms. On each sympodium, the first bract subtends a precocious axillary bud that reiterates the sympodial unit; the second bract subtends a bud that develops the horizontal shoot. In both horizontal and vertical shoots, the internode below the second bract is produced by both the second bract and the renewal shoot. Sympodial growth is present in seedlings. In other species of Eleocharis, the structure of the sympodial unit is conserved but morphological diversity develops from variation in horizontal shoot growth. PMID- 21642135 TI - Gas exchange and carbon metabolism in two Prosopis species (Fabaceae) from semiarid habitats: effects of elevated CO2, N supply, and N source. AB - Predicting future plant and ecosystem responses to elevated CO(2) also requires an understanding of the role of other factors, especially soil nitrogen. This is particularly challenging for global aridlands where total N and the relative amounts of nitrate and ammonia vary both spatially and seasonally. We measured gas exchange and primary and secondary C metabolites in seedlings of two dominant aridland shrub species (Prosopis flexuosa [S America] and P. glandulosa [N America]) grown at ambient (350 ppm) or elevated (650 ppm) CO(2) and nitrogen at two levels (low [0.8 mM] and high [8.0 mM]) and at either 1 : 1 or 3 : 1 nitrate to ammonia. Whereas elevated CO(2) increased assimilation rate, water use efficiency, and primary carbon metabolites in both species, these increases were strongly contingent upon nitrogen availability. Elevated CO(2) did not increase secondary metabolites (i.e., phenolics). For these important aridland species, the effects of elevated CO(2) are strongly influenced by nitrogen availability and to a lesser extent by the relative amounts of nitrate and ammonia supplied, which underscores the importance of both the amount and chemical composition of soil nitrogen in mediating the potential responses of seedling growth and establishment of aridland plants under future CO(2)-enriched atmospheres. PMID- 21642136 TI - Cataphylls of the Middle Triassic cycad Antarcticycas schopfii and new insights into cycad evolution. AB - Cataphylls associated with the Middle Triassic stem genus Antarcticycas are described, and their impact on understanding cycad evolution is discussed. The cataphylls of Antarcticycas are triangular in outline and flattened adaxially with lateral flanges. The outer surfaces are covered with a ramentum of filamentous hairs, the epidermis is a single cell layer thick, and the ground tissue is parenchymatous with mucilage canals and sclereids. Vascular bundles form a distinct inverted omega-shaped pattern characteristic of the Cycadales observed in petioles of extant species. The structures in Antarcticycas are interpreted as cataphylls based on overall morphology, presence of straight vascular strands in the cortex of the associated stem, and lack of fascicular cambia in the vascular bundles. Because much of the overall diversity of Cycadales is represented by fossils, integrating fossil taxa into explicit phylogenetic hypotheses is important for understanding cycad evolution. Therefore, character and minimum age mapping were performed on a phylogeny of extant and fossil taxa including Antarcticycas. The results suggest that major extant lineages of Cycadales had diverged by the Permian to Triassic and that certain synapomorphies for Cycadales had evolved by the Permian. Evidence of insect feeding on Antarcticycas suggests that associations between cycads and insects are ancient. PMID- 21642137 TI - Inbreeding effect on male and female fertility and inheritance of male sterility in Nemophila menziesii (Hydrophyllaceae). AB - Models of the evolution of gynodioecy assume that inbreeding affects male and female fertility equally and ignore quantitative variation in sex expression. The objectives of this study were to assess inbreeding effects, genetic background, and plant maturity on male and female fertility and the mechanism of male sterility inheritance for Nemophila menziesii (Hydrophyllaceae). Frequency of male-sterile flowers, number of anthers and ovules, and percentage of viable pollen were measured on plants from different pedigrees and five inbreeding levels (F = 0, 0.0625, 0.25, 0.5, and 0.75). Quantitative variation in male sterility was evident. As inbreeding increased, anther and ovule number decreased; the effect on anther number was greater than on ovule number. Pedigrees varied in number of male-sterile flowers and inbreeding effects. Frequency of male-sterile flowers was greatest among first flowers. No trade-off between male and female fertility was detected. A model attributing male sterility to a cytoplasmic locus and restoration to male fertility to a nuclear locus accounted for the distribution of complete sterility and hermaphroditism over the pedigrees. This study suggests that models of the evolution and maintenance of gynodioecy should allow for quantitative variation in male and female fertility components due to inbreeding, pedigree, and plant maturity. PMID- 21642138 TI - The evolutionary history of Sium sensu lato (Apiaceae): dispersal, vicariance, and domestication as inferred from ITS rDNA phylogeny. AB - The biogeographic history of Sium sensu lato (s.l.) (including Sium sensu stricto [s.s.], Afrocarum, and Berula) was inferred using a nuclear rDNA ITS sequence phylogeny and dispersal-vicariance analysis. One hundred accessions were analyzed, including 86 specimens of Sium s.l., representing all taxa throughout their geographical range. The phylogenetic analyses showed that Afrocarum and the African and St. Helena species of Sium were nested within Berula forming the Berula s.l. clade. The remaining species of Sium constituted a weakly supported Sium s.s. clade. The cultivated S. sisarum is closely related to the western Asian S. sisaroideum. The divergence of Sium s.s. from Berula s.l., estimated under the assumption of a molecular clock and using a substitution rate of 0.61% per million years, took place around 10.4 million years ago. Dispersal-vicariance analysis suggested that the biogeographic pattern of this group resulted from simple vicariance and eventual migration to adjacent areas rather than from long distance dispersal. The ancestral area of Sium s.l. was probably western Palearctic. The dispersal of Berula to North America occurred ca. 3 million years ago, possibly by island hopping. The migration of S. suave to North America took place ca 0.6 million years ago through Beringia. PMID- 21642139 TI - Alkaloid polymorphism and ITS sequence variation in the Spiraea japonica complex (Rosaceae) in China: traces of the biological effects of the Himalaya-Tibet Plateau uplift. AB - Spiraea japonica, a diverse, perennial shrubby species complex widespread across E Asia, was a useful model in a first attempt to link specific phylogeny, floristic evolution, and historical environmental changes in E Asia. DNA-based phylogenetic reconstruction served as a framework to investigate whether the patterns of variation in alkaloids and nrITS from varieties of S. japonica reflect the geological history of China and the uplifting of the Himalaya-Tibet plateau, the most significant geological event in E Asia since the late Paleocene. The high geographical structure found in the phylogenetic trees was corroborated by the distribution of alkaloids. All populations from E China, with mainly hetisine-type alkaloids, formed a single clade, sister to a clade of all populations from SW China, with mainly atisine-type alkaloids. The distribution boundary between the two clades roughly matched the floristic division between the Sino-Japanese Forest and the Sino-Himalayan Forest subkingdoms, as well as the environmental division between SW and E China, suggesting a close link between specific phylogeny, floristic evolution, and geographical changes in E Asia. The divergence between lineages at variety and population level within the eastern clade was slightly older than those within the southwestern clade, supporting the hypothesis of a northeast-southwest migration of Spiraea since the Eocene. The uplift of the Himalaya-Tibet plateau and subsequent increase in geographical complexity in SW China, could facilitate divergence maintenance, thus accelerating the evolutionary rate. PMID- 21642140 TI - Molecular phylogenetics of Vandeae (Orchidaceae) and the evolution of leaflessness. AB - Members of tribe Vandeae (Orchidaceae) form a large, pantropical clade of horticulturally important epiphytes. Monopodial leafless members of Vandeae have undergone extreme reduction in habit and represent a novel adaptation to the canopy environment in tropical Africa, Asia, and America. To study the evolution of monopodial leaflessness, molecular and structural evidence was used to generate phylogenetic hypotheses for Vandeae. Molecular analyses used sequence data from ITS nrDNA, trnL-F plastid DNA, and matK plastid DNA. Maximum parsimony analyses of these three DNA regions each supported two subtribes within monopodial Vandeae: Aeridinae and a combined Angraecinae + Aerangidinae. Adding structural characters to sequence data resulted in trees with more homoplasy, but gave fewer trees each with more well-supported clades than either data set alone. Two techniques for examining character evolution were compared: (1) mapping vegetative characters onto a molecular topology and (2) tracing vegetative characters onto a combined structural and molecular topology. In both cases, structural synapomorphies supporting monopodial Vandeae were nearly identical. A change in leaf morphology (usually reduced to a nonphotosynthetic scale), monopodial growth habit, and aeration complexes for gas exchange in photosynthetic roots seem to be the most important characters in making the evolutionary transition to leaflessness. PMID- 21642141 TI - An origin of aerial branch parasitism in the mistletoe family, Loranthaceae. AB - The large mistletoe family, Loranthaceae, contains 75 genera and approximately 1000 species. The family originated in the Southern Hemisphere and dispersed, apparently early, between fragments of Gondwana. It is now widely distributed on land surfaces of the former supercontinent. The Loranthaceae has three terrestrial, root-parasitic genera-a habit considered ancestral-and 72 genera of aerial, branch parasites. For almost two centuries, the origin of the mistletoe habit has been of interest to biologists. Two main evolutionary pathways have been proposed to explain the transition from terrestrial to aerial parasitism in the family. One theorizes the presence of an intermediate climbing ancestor in the path to the aerial habit. The other proposes a direct transfer from terrestrial to epiphytic growth following the germination of seeds on tree branches. Here we present molecular and morphological evidence that (1) the terrestrial species Nuytsia floribunda is ancestral within the Loranthaceae, (2) aerial parasitism has had multiple origins in the family, (3) the first aerial branch parasites had epicortical roots, and (4) the origin of aerial parasitism in one Old World clade involved the direct transfer from terrestrial to epiphytic growth following the germination of seeds on tree branches. Our results suggest that it is not necessary to evoke a climbing intermediate in the origins of aerial parasitism in the Santalales. PMID- 21642142 TI - A highly differentiated glomeromycotean association with the mucilage-secreting, primitive antipodean liverwort Treubia (Treubiaceae): clues to the origins of mycorrhizas. AB - Thallus anatomy in three species of the primitive liverwort genus Treubia (Metzgeriidae, Treubiales) was studied by light and electron microscopy. The thallus exudes copious mucilage, a feature shared elsewhere in liverworts only with the mycotrophic subterranean axes of the allied genus Haplomitrium. The central strand in the thallus midrib has a unique histological organization and harbors an intra- and intercellular infection by a glomeromycotean fungus that is far more highly differentiated than most of the glomeromycotean associations described to date. The fungus enters the thallus via clefts in the ventral epidermis along the midrib and colonizes the parenchyma above, forming intracellular coils and prominent, relatively short-lived, hyphal swellings. Above the zone with intracellular colonization is a tissue area containing mucilage-filled intercellular spaces; here the fungus is entirely intercellular and forms abundant pseudoparenchymatous structures and, in more mature parts of the thalli, large hyphae with thick multistratose walls. Mucilage in Treubia differs in histochemistry and origin from that produced by apical papillae, via hypertrophied Golgi, in all other bryophytes. Remarkable parallels between fungal associations in Treubia, Haplomitrium, and Lycopodium, all members of very ancient lineages, suggest that these associations epitomize very early stages in the evolution of glomeromycotean symbioses. PMID- 21642143 TI - Minisatellite telomeres occur in the family Alliaceae but are lost in Allium. AB - Although telomere sequences are considered to be highly conserved, there are switch-points in plant telomere evolution that are congruent with species' phylogenies. When Asparagales diverged, the Arabidopsis-type telomeric minisatellite repeat (TTTAGGG)(n) was first replaced by a human-type (TTAGGG)(n) repeat, and both were lost in Allium cepa (Alliaceae). We aimed to discover (1) when this loss occurred during divergence of Alliaceae and, (2) if (TTAGGG)(n) repeats were replaced by other known telomeric minisatellites. Slot-blot hybridization, fluorescent in situ hybridization, BAL31 digestion, asymmetric PCR, and cloning were used to identify and localize candidate telomeric sequences in species of Nothoscordum, Miersia, Ipheion, Tulbaghia, Gethyum, Gilliesia, Leucocoryne, Tristagma, and representatives of the three major Allium clades. Alliaceae genera other than Allium have human (TTAGGG)-type telomeric repeats that form telomeres. In Allium, only Tetrahymena-type (TTGGGG) repeats were ubiquitous in the genome, but they were not localized to telomeres. Likewise, the consensus telomeric repeats in Arabidopsis, human, Bombyx (TTAGG), Chlamydomonas (TTTTAGGG), and Oxytricha (TTTTGGGG) are absent in Allium telomeres. Alliaceae with human-type telomeres share telomere structures with related Asparagales species. We demonstrate that in the Allium ancestor human-type telomeric repeats were lost from telomeres and were not replaced by any investigated alternative minisatellite repeats. However, human and other types of minisatellite telomeric repeats are interspersed in some Allium genomes and their genomic signatures coincide with Allium clades. PMID- 21642144 TI - Allometric theory and the mechanical stability of large trees: proof and conjecture. AB - Recent allometric theory has postulated that standing leaf mass will scale as the 3/4 power of stem mass and as the 3/4 power of root mass such that stem mass scales isometrically with respect to root mass across very large vascular plant species with self-supporting stems. We show that the isometric scaling of stem mass with respect to root mass (i.e., M(S) ? M(R)) can be derived directly from mechanical theory, specifically from the requirement that wind-induced bending moments acting at the base of stems must be balanced by a counter-resisting moment provided by the root system to prevent uprooting. This derivation provides indirect verification of the allometric theory. It also draws attention to the fact that leaf, stem, and root biomass partitioning patterns must accommodate the simultaneous performance of manifold functional obligations. PMID- 21642145 TI - How strong is intracanopy leaf plasticity in temperate deciduous trees? AB - Intracanopy plasticity in tree leaf form is a major determinant of whole-plant function and potentially of forest understory ecology. However, there exists little systematic information for the full extent of intracanopy plasticity, whether it is linked with height and exposure, or its variation across species. For arboretum-grown trees of six temperate deciduous species averaging 13-18 m in height, we quantified intracanopy plasticity for 11 leaf traits across three canopy locations (basal-interior, basal-exterior, and top). Plasticity was pronounced across the canopy, and maximum likelihood analyses indicated that plasticity was primarily linked with irradiance, regardless of height. Intracanopy plasticity (the quotient of values for top and basal-interior leaves) was often similar across species and statistically indistinguishable across species for several key traits. At canopy tops, the area of individual leaves was on average 0.5-0.6 times that at basal-interior, stomatal density 1.1-1.5 times higher, sapwood cross-sectional area up to 1.7 times higher, and leaf mass per area 1.5-2.2 times higher; guard cell and stomatal pore lengths were invariant across the canopy. Species differed in intracanopy plasticity for the mass of individual leaves, leaf margin dissection, ratio of leaf to sapwood areas, and stomatal pore area per leaf area; plasticity quotients ranged only up to ~2. Across the six species, trait plasticities were uncorrelated and independent of the magnitude of the canopy gradient in irradiance or height and of the species' light requirements for regeneration. This convergence across species indicates general optimization or constraints in development, resulting in a bounded plasticity that improves canopy performance. PMID- 21642146 TI - Population divergence for heteroblasty in the Canary Island pine (Pinus canariensis, Pinaceae). AB - A heteroblastic (or vegetative phase) change is an abrupt manifestation in the general heteroblastic development during the ontogeny of plants. The Canary Island pine undergoes an especially marked and delayed heteroblastic change, including both the formation of secondary needles on dwarf shoots and the onset of preformed growth. To assess genetic and environmental effects on the heteroblastic change in this species, we followed plants from 19 populations at a dry site and a wetter site. Comparing juvenile and adult needles from the same individuals, the adult had a significantly lower rate of water loss and higher leaf mass per area. Pooling data from all seed sources, the heteroblastic change took place when plants reached a critical height, on average, at 4 years of age at the dry site and 1 year earlier at the wet site. Within a subsample of individuals of equal size, mortality was significantly higher in juvenile plants than in mature plants. However, the juvenile phase was longer in plants from dry regions when compared to plants from highly productive, wet regions. This apparent contradiction might be explained through differential resource allocation and the cost of sclerophylly and resprouting ability. Considering the life strategy of the Canary Island pine, we interpret the prolonged juvenile phase as an unavoidable trade-off for the high tolerance of adults to harsh environments. PMID- 21642147 TI - Post-dispersal embryo development, germination phenology, and seed dormancy in Cardiocrinum cordatum var. glehnii (Liliaceae s. str.), a perennial herb of the broadleaved deciduous forest in Japan. AB - In an investigation of seed germination in Cardiocrinum cordatum var. glehnii, embryos in fresh seeds in October were underdeveloped and did not grow until September of the following year. Then, they grew rapidly and had fully elongated by early November. In the second spring after dispersal, radicles emerged under snow in late March and after snowmelt in April. Cotyledons emerged soon after radicles. In several laboratory experiments, embryos grew at 15 degrees /5 degrees C (light 12 h/ dark 12 h) following 25 degrees /15 degrees C. Radicles emerged from seeds with fully elongated embryos at 5 degrees -15 degrees C after cold stratification at 0 degrees -5 degrees C. Cotyledons emerged in 2 wk from seeds with a radicle at 15 degrees /5 degrees C to 30 degrees /20 degrees C. Although seeds require c. 18-19 mo after dispersal to germinate in nature, under controlled conditions, they required only 9 mo with a sequence of 25 degrees /15 degrees C -> 15 degrees /5 degrees C -> 0 degrees -5 degrees C -> 15 degrees /5 degrees C. This is practical knowledge for propagation of plants from seeds. GA(3) treatment partially substituted for the high temperature requirement. Based on dormancy-breaking requirements, the seeds have deep simple morphophysiological dormancy (MPD). A literature review of seed dormancy in taxa of Liliaceae s. str. showed that phylogenetic position in this case is not a good predictor of level of MPD. PMID- 21642148 TI - Relaxed pollinator-mediated selection weakens floral integration in self compatible taxa of Leavenworthia (Brassicaceae). AB - Natural selection should favor the integration of floral traits that enhance pollen export and import in plant populations that rely upon pollinators. If this is true, then phenotypic correlations between floral traits should weaken in self fertilizing groups that do not require pollinator visitation to produce seed. We tested this hypothesis in Leavenworthia, a plant genus in which there have been multiple independent losses of the sporophytic self-incompatibility system found throughout the Brassicaceae. In particular, we conducted phylogenetically independent contrasts of floral trait correlations between two pairs of self incompatible (SI) and self-compatible (SC) sister taxa. In support of the hypothesis that pollinator-mediated selection integrates floral traits, we found that both SC Leavenworthia taxa have weaker overall floral correlations in comparison to sister taxa that rely upon pollinators. The two independently derived SC Leavenworthia flowers have significantly weaker stamen-petal or pistil petal correlations, respectively, whereas the stamen-pistil correlation remains constant. These patterns suggest that relaxation of pollinator-mediated selection weakens the integration of traits associated with pollen export and import. The retention of high stamen-pistil correlations in the SC taxa of Leavenworthia further implies that the integration of these traits is either constrained or maintained by selection favoring the successful transfer of pollen within flowers to secure self-pollination. PMID- 21642149 TI - Effects of an exotic plant and habitat disturbance on pollinator visitation and reproduction in a boreal forest herb. AB - The invasion of exotic species into natural habitats is considered to be a major threat to biodiversity, and many studies have examined how exotic plants directly affect native plant species through competitive interactions for abiotic resources. However, although exotics can have potentially great ecological and evolutionary consequences, very few researchers have studied the effect of exotics on the interactions between plants and their mutualistic partners, such as pollinators, and none have reported on such impacts in logged and undisturbed boreal forest ecosystems. Here we show how experimental introductions of an exotic plant species (Phacelia tanacetifolia Bentham) affect pollinator visitation and female reproductive success of a native plant (Melampyrum pratense L.) in recently disturbed (i.e., logged) and in undisturbed boreal forest habitats. The presence of Phacelia significantly increased the number of bumble bees entering plots in both habitat types. However, the exotic species had a strong negative impact on the visitation rate to the native species in both habitat types. Despite this negative impact on pollinator visitation, the exotic had no effect on female reproductive success of the native species in any habitat. Our results show that seed production may be more robust than pollinator visitation to exotic invasion, irrespective of habitat disturbance history. PMID- 21642150 TI - A genetic appraisal of a new synthetic Nicotiana tabacum (Solanaceae) and the Kostoff synthetic tobacco. AB - Polyploids have significantly influenced angiosperm evolution. Understanding the genetic consequences of polyploidy is advanced by studies on synthetic allopolyploids that mimic natural species. In Nicotiana, Burk (1973) and Kostoff (1938) generated synthetic tobacco (N. tabacum) using the parents ?N. sylvestris * ?N. tomentosiformis. We previously reported rapid genetic changes in the Burk material. Kostoff's material has 24 chromosomes of N. sylvestris origin (S genome), 24 of N. tomentosiformis origin (T-genome), and a large intergenomic translocation, but not an additive distribution of ribosomal DNA (rDNA) families as expected from the parental contribution. Our new synthetic tobacco lines TR1 and TR2 are chromosomally balanced with no intergenomic translocations and are either sterile or have highly reduced fertility, supporting the nuclear cytoplasmic hypothesis that allopolyploid fertility is enhanced by intergenomic translocations. Two plants of TR1 (TR1-A, TR1-B) have the expected number, structure, and chromosomal distribution of rDNA families, in contrast to Burk's and Kostoff's synthetic tobaccos and to synthetic polyploids of Arabidopsis. Perhaps allopolyploids must pass through meiosis before genetic changes involving rDNA become apparent, or the genetic changes may occur stochastically in different synthetic allopolyploids. The lack of fertility in the first generation of our synthetic tobacco lines may have uses in biopharmacy. PMID- 21642151 TI - Vascular pathways constrain 13C accumulation in large root sinks of Lycopersicon esculentum (Solanaceae). AB - While carbon transport and partitioning is largely determined by phloem source sink relationships, it may be constrained by vascular connections. Tomato (Lycopersicon esculentum) plants exhibit a high degree of sectoriality, with restricted movement of nutrients from particular roots to orthostichous leaves. In this experiment we investigated the manner in which sectoriality influences source-sink phloem partitioning from shoots to roots in tomatoes and whether the size of the sink (root) modifies the pattern of carbon movement outside sectored pathways. Using (13)C, we determined that shoot-to-root carbon transport in tomatoes is sectored even from upper leaves. Sink size also influenced carbon partitioning. Specifically, when a lateral root was grown in isolation (using a split-pot technique), it grew more and acquired significantly more (13)C from an orthostichous, exposed leaf than did any other single root. Vascular constraints were evident. (13)C accumulation in a large, isolated lateral root was very low when a leaf opposite the isolated lateral root was exposed. Thus sink size did not overcome vascular constraints. Because carbon assimilates are needed for nutrient acquisition and assimilation, these vascular constraints may affect the ability of sectored plants to utilize heterogeneously distributed soil resources. If so, future studies should compare species that differ in sectoriality to determine whether vascular constraints affect competitive hierarchies when soil resource availability is patchy. PMID- 21642152 TI - A stochastic population model for Lepidium papilliferum (Brassicaceae), a rare desert ephemeral with a persistent seed bank. AB - Population viability analysis (PVA) is a valuable tool for rare plant conservation, but PVA for plants with persistent seed banks is difficult without reliable information on seed bank processes. We modeled the population dynamics of the Snake River Plains ephemeral Lepidium papilliferum using data from an 11 yr artificial seed bank experiment to estimate age-specific vital rates for viability loss and germination. We related variation in postgermination demographic parameters to annual variation in precipitation patterns and used these relationships to construct a stochastic population model using precipitation driver variables. This enabled us to incorporate realistic levels of environmental variability into the model. A model incorporating best estimates for parameter values resulted in a mean trajectory for seed bank size that remained essentially stable through time, although there was a measurable risk of extinction over a 100-yr period for the study population under this scenario. Doubling the annual seed viability loss rate resulted in near-certain extinction, as did increasing first-year germination to 100%, showing the importance of the persistent seed bank. Interestingly, increasing environmental variance substantially decreased the risk of extinction, presumably because this plant relies on extremely good years to restock the persistent seed bank, while extremely bad years have little impact. If every year were average in this desert environment, the species could not persist. Simulated effects of livestock trampling resulted in greatly increased extinction risk, even over time frames as short as 15 years. PMID- 21642153 TI - Asymmetrical pollen success in Ipomopsis (Polemoniaceae) contact sites. AB - Variation in hybridization rates among contact sites of a species pair provides an opportunity for assessing the importance of individual reproductive isolating mechanisms in limiting gene flow between species and thus promoting speciation. Conspecific pollen advantage is common in angiosperms, but its importance as a reproductive isolating mechanism is uncertain. We compared the strength of conspecific pollen advantage in two Ipomopsis aggregata-I. tenuituba (Polemoniaceae) contact sites that differ in frequency of natural hybrids. We performed hand pollinations of single- and 1:1 mixed-species pollen loads, using donor and recipient plants from both contact sites. Paternity of offspring from mixed-species pollinations was determined using an allozyme marker. Donors from the high frequency hybrid site showed no conspecific pollen advantage; both species sired seeds in proportion to their fraction of the pollen load (0.5). In contrast, I. aggregata from the low frequency hybrid site sired 70-85% of offspring on recipients from both sites. These results suggest that pollen interactions can influence the level of natural hybridization. They also suggest the importance of geographic variation in reproductive isolation, which should be considered in studies of biological invasions and exposure of engineered crops to wild relatives. PMID- 21642154 TI - Fruit production in cranberry (Ericaceae: Vaccinium macrocarpon): a bet-hedging strategy to optimize reproductive effort. AB - In the cultivated cranberry (Vaccinium macrocarpon), reproductive stems produce 1 3 fruit even though they usually have 5-7 flowers in the spring. We undertook experiments to test the hypothesis that this was an adaptive life history strategy associated with reproductive effort rather than simply the result of insufficient pollination. We compared fruit production on naturally pollinated plants with those that were either manually pollinated or that were caged to exclude insects. Clearly, insects are necessary for the effective pollination of cranberry plants, but hand pollination of all flowers did not result in an increase in fruit number. Most of the upper flowers, which had significantly fewer ovules than did the lower flowers, aborted naturally soon after pollination. However, when the lower flower buds were removed, the upper flowers produced fruit. This suggests that the upper flowers may serve as a backup if the earlier blooming lower ones are lost early in the season. Furthermore, the late blooming flowers may still contribute to the plant's reproductive success as visiting pollinators remove the pollen, which could serve to sire fruit on other plants. These results are discussed in the context of their possible evolutionary and proximate causes. PMID- 21642155 TI - Floral syndromes accurately predict pollination by a specialized oil-collecting bee (Rediviva peringueyi, Melittidae) in a guild of South African orchids (Coryciinae). AB - The long-standing notion of pollination syndromes, which postulates that plants form recognizable groups according to pollinator type, has been challenged recently on the basis of apparent widespread generalization in pollination systems. As a test of the pollination syndrome concept, I examined the pollination biology of a group of 15 orchids that share a recognizable syndrome of floral features that includes yellow-green coloration, oil secretion, pungent scent, shallow flowers, and a September peak in flowering. The orchids occur in sympatry in the Cape Floral Region of South Africa. According to the pollination syndrome concept, the similar floral features of this group indicate a shared pollinator. To test this prediction, I observed pollinators on Pterygodium alatum, P. caffrum, P. catholicum, P. volucris, Corycium orobanchoides, and Disperis bolusiana subsp. bolusiana. They shared a single species of pollinator, the oil-collecting bee, Rediviva peringueyi. Female bees collected oil from the lip appendage using modified front tarsi. The orchids reduce interspecific reproductive interference through differences in pollinarium length or the use of mutually exclusive pollinarium attachment sites on the body of the bee. The results are contrary to the expectation of generalization in pollination systems and suggest that pollinators play an important role in mediating selection on floral traits. PMID- 21642156 TI - Introduction of Glossostigma (Phrymaceae) to North America: a taxonomic and ecological overview. AB - Field surveys in eastern North America confirm the naturalization of Glossostigma plants at 19 localities in four states: Connecticut, New Jersey, Pennsylvania, and Rhode Island. DNA sequence analysis of individuals from 14 sampled populations identifies these nonindigenous plants as Glossostigma cleistanthum, a species native to Australia and New Zealand. These results correct prior misidentifications of North American plants as G. diandrum. The earliest North American record of G. cleistanthum (1992) is from a Ramsar tidal wetland in Connecticut. Morphological analyses demonstrate that G. cleistanthum differs from G. diandrum by its longer leaves and ability to produce both cleistogamous and chasmogamous flowers in response to ecological conditions. Glossostigma cleistanthum has a high reproductive potential and spreads rapidly within and between both artificial and natural habitats. A survey of more than 100 lakes indicated that G. cleistanthum occurs most often in waters with high clarity and low pH, alkalinity, conductivity, and phosphorous. Because of its affinity for oligotrophic conditions, this species is a particular threat to pristine natural aquatic communities, which often contain imperiled plants. PMID- 21642157 TI - From the first century of the botanical society of america into the second. PMID- 21642158 TI - One hundred years of American botany: a short history of the Botanical Society of America. AB - This paper offers highlights from the 100 (plus) years of the Botanical Society of America (BSA) and draws extensively on the archives of the BSA. In addition to examining the founding of the society and the attempt to "professionalize" botany in late 19th century America, the paper also explores the complex relations between the BSA and a number of related societies in the United States, the Society's struggle to create a coherent identity for itself, the place of botany as a whole in the context of the burgeoning biological sciences in the 20th century, and the changing role of the BSA in an international context. The paper assesses both the achievements and the challenges facing the BSA. It closes by offering some historical reflections on the status of "botany" as a science and the historical significance of terms like "plant biology" and "plant science." PMID- 21642159 TI - The molecular genetic basis of plant adaptation. AB - How natural selection on adaptive traits is filtered to the genetic level remains largely unknown. Theory and quantitative trait locus (QTL) mapping have provided insights into the number and effect of genes underlying adaptations, but these results have been hampered by questions of applicability to real biological systems and poor resolution, respectively. Advances in molecular technologies have expedited the cloning of adaptive genes through both forward and reverse genetic approaches. Forward approaches start with adaptive traits and attempt to characterize their underlying genetic architectures through linkage disequilibrium mapping, QTL mapping, and other methods. Reverse screens search large sequence data sets for genes that possess the signature of selection. Though both approaches have been successful in identifying adaptive genes in plants, very few, if any, of these adaptations' molecular bases have been fully resolved. The continued isolation of plant adaptive genes will lead to a more comprehensive understanding of natural selection's effect on genes and genomes. PMID- 21642160 TI - Origins and nature of vessels in monocotyledons: 8. Orchidaceae. AB - Xylem of the orchids studied provided unusually favorable material to demonstrate how conductive tissue evolves in monocotyledons. In the end walls of tracheary elements of many Orchidaceae, remnants of pit membranes were observed with scanning electron microscopy and minimally destructive methods. The full range from tracheids to vessel elements, featuring many intermediate stages, was illustrated with SEM in hand sections of fixed roots, stems, and inflorescence axes of 13 species from four subfamilies. Pit membranes in end walls of tracheary elements are porose to reticulate in roots of all species, but nonporose in stems of Cypripedioideae and Vanilloideae and porose to reticulate in stems of Orchidoideae and Epidendroideae. The distribution pattern of pit membranes and pit membrane remnants in end walls of tracheary elements of orchids parallels the findings of others. The position of Cypripedioideae and Vanilloideae as outgroups to Orchidoideae and Epidendroideae, claimed by earlier authors, is supported by clades based on molecular studies and by our studies. Little hydrolysis of pit membranes in tracheary element end walls was observed in pseudobulbs or inflorescence axes of epidendroids. The pervasiveness of network-like pit membranes of various extents and patterns in end walls of tracheary elements in Orchidaceae calls into question the traditional definitions of tracheids and vessel elements, not merely in orchids, but in angiosperms at large. These two concepts, based on light microscope studies, are blurred in light of ultrastructural studies. More importantly, the intermediate expressions of pit membranes in tracheary element end walls of Orchidaceae and some other families of angiosperms are important as indicators of steps in evolution of conduction with respect to organs (more rapid flow in roots than in succulent storage structures) and habitat (less obstruction to flow correlated with a shift from terrestrial to epiphytic). PMID- 21642161 TI - Differential responses of invasive Celastrus orbiculatus (Celastraceae) and native C. scandens to changes in light quality. AB - When plants are subjected to leaf canopy shade in forest understories or from neighboring plants, they not only experience reduced light quantity, but light quality in lowered red : far red light (R : FR). Growth and other developmental responses of plants in reduced R : FR can vary and are not consistent across species. We compared how an invasive liana, Celastrus orbiculatus, and its closely related native congener, C. scandens, responded to changes in the R : FR under controlled, simulated understory conditions. We measured a suite of morphological and growth attributes under control, neutral shading, and low R : FR light treatments. Celastrus orbiculatus showed an increase in height, aboveground biomass, and total leaf mass in reduced R : FR treatments as compared to the neutral shade, while C. scandens had increased stem diameter, single leaf area, and leaf mass to stem mass ratio. These differences provide a mechanistic understanding of the ability of C. orbiculatus to increase height and actively forage for light resources in forest understories, while C. scandens appears unable to forage for light and instead depends upon a light gap forming. The plastic growth response of C. orbiculatus in shaded conditions points to its success in forested habitats where C. scandens is largely absent. PMID- 21642162 TI - A stage-based study of drought response in Cryptantha flava (Boraginaceae): gas exchange, water use efficiency, and whole plant performance. AB - Models of global climate change predict an increase in the frequency of major droughts, yet we know little about the consequences of drought for the demography of natural populations. This study examined a population of the semi-desert perennial Cryptantha flava (Boraginaceae) to determine how plants of different developmental stages respond to drought through changes in leaf gas exchange, leaf water potential, water use efficiency, growth, and reproduction. In two of the four years, drought was applied using rainout shelters, and a severe natural drought occurred in another. Small, presumably younger, plants sometimes had lower rates of maximum photosynthesis, lower leaf water potentials, and lower instantaneous or integrated water-use efficiency than large plants. Small plants also had higher relative growth rates and lower reproductive effort. Large plants with evidence of shrinkage from a previously larger size often produced less growth and reproduction than large healthy plants, suggesting a decline in plant vigor with age. Drought depressed gas exchange and leaf water potentials equally in all plant stages. Thus, leaf-level physiological attributes provide no clues for why drought reduces growth more strongly in large plants. The results point to several additional avenues of research relevant to understanding stage dependent or age-dependent plant performance under drought conditions. PMID- 21642163 TI - Do sexual dimorphisms in reproductive allocation and new shoot biomass increase with an increase of altitude? A case of the shrub willow Salix reinii (Salicaceae). AB - Based on the general tendency for females of dioecious plants to pay higher reproductive cost than males, it has been predicted that females should have much more reduced reproductive outputs and diminished vegetative production than males in energy-limited habitats. Nevertheless, this prediction has rarely been directly investigated. We investigated altitudinal changes in reproductive biomass and shoot production, normalized by plant size, for females and males of a shrub willow, Salix reinii, on Mt. Hakkoda, northeast Japan. Females maintained higher reproductive biomass than males at all altitudes; however, reproductive allocation for both sexes tended to decrease at a similar rate with an increase in altitude. Moreover, females vegetatively produced at the same rate as males at all altitudes. These findings suggest that females have a mechanism to compensate for the extra investment in reproduction irrespective of a changing environment. Shoot production did not change with altitude, suggesting that S. reinii gave priority to vegetative investment at the cost of reproductive output at higher altitudes. Inconsistent with general predictions, females did not respond more sensitively than males to severe environmental conditions in either reproductive allocation or shoot production, despite much higher resource investment in reproduction. PMID- 21642164 TI - Direct measurements of intervessel pit membrane hydraulic resistance in two angiosperm tree species. AB - The hydraulic resistance of pit membranes was measured directly in earlywood vessels of Fraxinus americana and Ulmus americana. The area-specific resistance of pit membranes (r(mem)) was higher than modeled or measured values obtained previously for hardwood species, with r(mem) of 5.24 * 10(3) MPa.s.m(-1) for Fraxinus and 2.56 * 10(3) MPa.s.m(-1) for Ulmus. The calculated resistance of pit canals was three orders of magnitude below total pit resistance indicating that pit membranes contributed the majority of resistance. Scanning electron microscopy indicated that pit membranes of Ulmus were thinner and more porous than those of Fraxinus, consistent with the difference in r(mem) between the species. Measurements of average vessel diameter and length and area of wall overlap with neighboring vessels were used to partition the vascular resistance between vessel lumen and pit membrane components. Pit membrane resistance accounted for 80% of the total resistance in Fraxinus and 87% in Ulmus in 2-yr old branch sections. However, measurements of vessel dimensions in the trunk suggest that the division of resistance between pit membrane and lumen components would be closer to co-limiting in older regions of the tree. Thus, pit membrane resistance may be of greater relative importance in small branches than in older regions of mature trees. PMID- 21642165 TI - Spatial genetic structure and frequency of interspecific hybridization in Platanthera aquilonis and P. dilatata (Orchidaceae) occurring in sympatry. AB - Knowledge of genetic structure at different scales is necessary for evaluating the importance of interactions between the genome and environment and for inferring underlying processes that bring about evolutionary diversification. Here, genetic and morphological variation was assessed for 154 individuals of Platanthera aquilonis and P. dilatata in Maine, using RAPD and PCR-RFLP markers and measurements of five morphological traits to determine the prevalence of interspecific hybrids and underlying spatial genetic structure of the population. Both species co-mingled in the population, but clumping was observed. Platanthera aquilonis was more abundant, but genetic variation was lower (polymorphic loci [40%], expected heterozygosity [0.137]) than that of P. dilatata (P(p) = 72%; H(e) = 0.245). Interspecific hybrids were rare (eight individuals), and morphology was not consistently reliable for determining hybrid status even though the species differed significantly in several traits. Spatial autocorrelation analyses showed significant genetic structure at small distances for both species, most likely due to restricted seed dispersal. Platanthera aquilonis did not exhibit a pattern of spatial genetic structure consistent with other selfing species. This suggests that the species is facultative autogamous, which allows for limited production of hybrid individuals and reduces the strength of spatial genetic structure relative to P. dilatata. PMID- 21642166 TI - Floral display and mating patterns within populations of the neotropical epiphytic orchid,Laeliarubescens (Orchidaceae). AB - Pollinator behavior plays a central role in determining patterns of pollen mediated gene movement in zoophilous angiosperms. A species' floral display can strongly influence the behavior of its pollinators and thereby affect its evolutionary pathway. We used paternity analysis to directly measure and describe mating patterns within 15 populations of the epiphytic orchid, Laelia rubescens, in Costa Rican dry forest. Strict correlated mating by orchids allows inference of the precise multilocus diploid genotype of the pollen parents. Our data show that mean effective population sizes were small (11.2 in 1999 and 11.8 in 2000) relative to the number of flowering genets (63 and 56, respectively). Fewer genets were reproductively successful as females than males. The relationship between reproductive success (RS) and floral display within three cluster size classes was consistent between years, with large (>30 inflorescences) and small (<=10 inflorescences) clusters often having significantly lower RS than expected, while the RS of medium-sized clusters (11-30 inflorescences) often significantly exceeded expectations. Paternity analysis allowed us to take advantage of the pollination biology of L. rubescens to provide unusually detailed insights into mating patterns, pollen-mediated gene movement and RS for populations of this epiphytic orchid, an herbaceous perennial, distributed in three-dimensional space. PMID- 21642167 TI - Population age structure and reproductive behavior of the monocarpic perennial Heracleum mantegazzianum (Apiaceae) in its native and invaded distribution ranges. AB - Many invasive species are benign in their native region-are there interactions between their key traits and the new habitats that explain invasion success? The giant perennial herb Heracleum mantegazzianum is a problematic invader in Europe and is also naturalized in North America. We compared its population structure and reproductive behavior in the native (W. Caucasus) and invaded (Czech Republic) areas in managed (pastures) and unmanaged sites. The age structure of the populations and age at flowering were analyzed using herb-chronology, a method based on counting annual rings in the secondary xylem of roots. The species was strictly monocarpic; most plants in unmanaged sites in the invaded range flowered in the third and fourth yr (maximum 12 yr). In unmanaged habitats, plants from the native range flowered later than those from the invaded range. In both ranges, flowering was delayed in managed sites where the population density was higher and most plants flowered around the fifth year. Reproductive output of individual plants was neither related to population density nor to age at flowering. More favorable climatic conditions in the invaded region, together with increased chances for dispersal in a densely colonized central Europe, seemed to allow the massive invasion. PMID- 21642168 TI - Ancestral reconstruction of flower morphology and pollination systems in Schizanthus (Solanaceae). AB - Concerted changes in flower morphology and pollinators provide strong evidence on adaptive evolution. Schizanthus (Solanaceae) has zygomorphic flowers and consists of 12 species of annual or biennial herbs that are distributed mainly in Chile and characterized by bee-, hummingbird-, and moth-pollination syndromes. To infer whether flowers diversified in relation to pollinator shifts, we traced the evolutionary trajectory of flower traits and visitors onto a phylogeny based on sequence data from ITS, waxy, and trnF/ndhJ DNA. Maximum-likelihood ancestral reconstruction of floral traits suggests that ancestral Schizanthus had a bee pollination syndrome. The hummingbird syndrome evolved in S. grahamii, a high elevation species in the Andes. The moth syndrome evolved in the ancestor of three species that inhabit the Atacama Desert. Results of mapping flower visitors onto the phylogeny show that the shift from bee to hummingbird pollination concurred with a shift in pollinators as predicted by the syndromes. However, the same pattern was not found for the moth syndrome. Visits by moths were observed only in one of the three moth-syndrome species, and at a very low rate. This mismatch suggests either anachronic floral characters or maintenance of rare, imperceptible moth pollination backed up by capacity for autonomous selfing. Overall, results suggest that diversification of flower traits in Schizanthus has occurred in relation to pollinator shifts. PMID- 21642169 TI - Phylogeny of the parasitic plant family Orobanchaceae inferred from phytochrome A. AB - Partial sequences of the nuclear gene encoding the photoreceptor phytochrome A (PHYA) are used to reconstruct relationships within Orobanchaceae, the largest of the parasitic angiosperm families. The monophyly of Orobanchaceae, including nonphotosynthetic holoparasites, hemiparasites, and nonparasitic Lindenbergia is strongly supported. Phytochrome A data resolve six well-supported lineages that contain all of the sampled genera except Brandisia, which is sister to the major radiation of hemiparasites. In contrast to previous plastid and ITS trees, relationships among these major clades also are generally well supported. Thus, the robust phylogenetic hypothesis inferred from the PHYA data provides a much better context in which to evaluate the evolution of parasitism within the group. Ninety-eight species of Orobanchaceae, representing 43 genera, are included and Brandisia, Bungea, Cymbaria, Esterhazya, Nesogenes, Phtheirospermum, Radamaea, Siphonostegia, and Xylocalyx are confirmed as members of Orobanchaceae. The earliest diverging lineage of hemiparasites is identified for the first time; it contains Bungea, Cymbaria, Monochasma, Siphonostegia, and the monotypic Schwalbea, which is federally endangered. This basal clade is marked by the presence of two novel introns. A second, apparently independent gain of one of these introns marks a clade of largely European taxa. There is significant rate heterogeneity among PHYA sequences, and the presence of multiple PHYA in some taxa is consistent with observed ploidy levels. PMID- 21642170 TI - The evolutionary history of Melianthus (Melianthaceae). AB - The evolutionary origins of the morphological and taxonomic diversity of angiosperms is poorly known. We used the genus Melianthus to explore the diversification of the southern African flora. Melianthus comprises eight species, and a phylogeny based on one nuclear and two plastid genes, as well as a morphological data set, confirmed that the genus is monophyletic. The two earliest diverging lineages are found in relatively mesic habitats, whereas the two terminal clades (an eastern and a western clade), each with three species, favor more arid habitats. The eastern clade is largely restricted to the summer rainfall parts of southern Africa, and the western clade is found in winter rainfall region. Molecular dating indicates a mid-Tertiary origin of the genus, with diversification of the eastern and western clades coincident with the Late Miocene-Pliocene uplift of the Escarpment mountains and the establishment of summer aridity along the west coast. The remarkably complex flowers are indicative of sunbird pollination, but many smaller birds can also visit. Speciation may be the consequence of allopatric divergence into edaphic-climatic niches. Divergence in flower and inflorescence morphology might be in response to the divergent pressures for nectar conservation in arid regions coupled with the need for signaling to avian pollinators in generally shrubby vegetation. PMID- 21642172 TI - Online errata. PMID- 21642171 TI - Homoplasious character combinations and generic delimitation: a case study from the Indo-Pacific arecoid palms (Arecaceae: Areceae). AB - The complex distributions of morphological character states in the Indo-Pacific palm tribe Areceae (Arecaceae; Arecoideae) are potentially challenging for the delimitation of its genera. In the first exhaustive sampling of all 65 genera of the Areceae, we examined relationships of two of the tribe's most problematic genera, Heterospathe and Rhopaloblaste, using portions of the low-copy nuclear genes phosphoribulokinase (PRK) and RNA-polymerase II subunit B (RPB2). Both genera fell within a highly supported clade comprising all Areceae genera, but are clearly unrelated. Rhopaloblaste was strongly supported as monophyletic and is most closely related to Indian Ocean genera. Heterospathe was resolved with strong support within a clade of western Pacific genera, but with the monotypic Alsmithia nested within it. Ptychosperma micranthum, which has previously been included in both Heterospathe and Rhopaloblaste, is excluded from these and from Ptychosperma, supporting its recent placement in a new genus Dransfieldia. Morphological comparisons indicate that the crownshaft is putatively synapomorphic for the Areceae with numerous reversals within the clade and some independent origins elsewhere. The putative diagnostic characters of Heterospathe show high levels of homoplasy, and the genus can only be distinguished by a suite of characters, whereas Rhopaloblaste is more clearly defined. Our results have implications not only for the two genera in focus, but have also been influential for the new classification of the Areceae. PMID- 21642173 TI - The pollination biology of Burmeistera (Campanulaceae): specialization and syndromes. AB - The floral traits of plants with specialized pollination systems both facilitate the primary pollinator and restrict other potential pollinators. To explore interactions between pollinators and floral traits of the genus Burmeistera, I filmed floral visitors and measured pollen deposition for 10 species in six cloud forest sites throughout northern Ecuador. Nine species were primarily bat pollinated (84-100% of pollen transfer); another (B. rubrosepala) was exclusively hummingbird-pollinated. According to a principal components analysis of 11 floral measurements, flowers of B. rubrosepala were morphologically distinct. Floral traits of all species closely matched traditional ornithophilous and chiropterophilous pollination syndromes; flowers of B. rubrosepala were bright red, lacked odor, opened in the afternoon, and had narrow corolla apertures and flexible pedicels, which positioned them below the foliage. Flowers of the bat pollinated species were dull-colored, emitted odor, opened in the evening, and had wide apertures and rigid pedicels, which positioned them beyond the foliage. Aperture width appeared most critical to restricting pollination; hummingbirds visited wide flowers without contacting the reproductive parts, and bats did not visit the narrow flowers of B. rubrosepala. Aperture width may impose an adaptive trade-off that favors the high degree of specialization in the genus. Other floral measurements were highly variable amongst bat-pollinated species, including stigma exsertion, calyx lobe morphology, and pedicel length. Because multiple species of Burmeistera often coexist, such morphological diversity may reduce pollen competition by encouraging pollinator fidelity and/or spatially partitioning pollinator's bodies. PMID- 21642174 TI - Phenotypic plasticity and mechanical stress: biomass partitioning and clonal growth of an aquatic plant species. AB - Mechanical stresses from wind, current or wave action can strongly affect plant growth and survival. Survival and distribution of species often depend on the plant's capacity to adapt to such stresses, particularly when amplified by climatic variations. Few studies have dealt with plastic adjustments in response to mechanical stress compared to resource stress. We hypothesized that mechanical stress should favor plastic adjustments that result in increased biomass production in zones protected from the stress and that altered growth patterns should be reversible after mechanical stress removal. Here we measured plastic adjustments in morphological traits and clonal architecture for an aquatic clonal species (Berula erecta) under two contrasting mechanical stresses in the field standing vs. running water. Reversion of the morphological changes was then assessed using transplants in standing water. In the case of mechanical stress, size reduction, biomass reallocation within clones (higher allocations to clonal growth and to belowground organs), and a more compact growth form (reduced spacer lengths) contributed to reducing the damage risk. The removal of mechanical stress induced compensatory growth, probably linked to the production of low density tissues. However, most patterns of dry mass partitioning induced by current stress were not reversed after stress removal. PMID- 21642175 TI - Assessing the use of the mitochondrial cox1 marker for use in DNA barcoding of red algae (Rhodophyta). AB - The red algae, a remarkably diverse group of organisms, are difficult to identify using morphology alone. Following the proposal to use the mitochondrial cytochrome c oxidase subunit I (cox1) for DNA barcoding animals, we assessed the use of this gene in the identification of red algae using 48 samples plus 31 sequences obtained from GenBank. The data set spanned six orders of red algae: the Bangiales, Ceramiales, Corallinales, Gigartinales, Gracilariales and Rhodymeniales. The results indicated that species could be discriminated. Intraspecific variation was between 0 and 4 bp over 539 bp analyzed except in Mastocarpus stellatus (0-14 bp) and Gracilaria gracilis (0-11 bp). Cryptic diversity was found in Bangia fuscopurpurea, Corallina officinalis, G. gracilis, M. stellatus, Porphyra leucosticta and P. umbilicalis. Interspecific variation across all taxa was between 28 and 148 bp, except for G. gracilis and M. stellatus. A comparison of cox1 with the plastid Rubisco spacer for Porphyra species revealed that it was a more sensitive marker in revealing incipient speciation and cryptic diversity. The cox1 gene has the potential to be used for DNA barcoding of red algae, although a good taxonomic foundation coupled with extensive sampling of taxa is essential for the development of an effective identification system. PMID- 21642176 TI - Allelochemic function for a primary metabolite: the case of l-tyrosine hyper production in Inga umbellifera (Fabaceae). AB - Young leaves of tropical forest trees experience far higher herbivory pressure than mature leaves of the same species. Selection on young leaves has led to diverse forms of defense chemical expression. Though most allelochemicals are secondary metabolites, allelochemic function for a primary metabolite remains a possibility. We recently observed this phenomenon in the young leaves of Inga umbellifera, which accumulate the protein amino acid l-tyrosine to very high levels. We isolated l-tyrosine from young leaves of trees in Panama and characterized it using spectroscopic and chemical means. We chromatographically quantified leaf l-tyrosine levels across a range of developmental stages, showing that it was present in the youngest leaves and that its concentration increased throughout the period of expansion, reaching an average maximum of ca 10% of leaf dry mass in late-stage young leaves. This chemical phenotype was seen to be highly leaf-age specific: Free tyrosine was only present in mature leaves at very low levels. In bioassays with larvae of the noctuid moth H. virescens, l-tyrosine proved to be a potent growth inhibitor when added to artificial diet at 10% of dry mass. This suggests that a rarely observed defense strategy occurs in young I. umbellifera leaves, a hyper-produced primary metabolite functioning as an allelochemical. PMID- 21642177 TI - Patterns of genetic divergence of three Canarian endemic Lotus (Fabaceae): implications for the conservation of the endangered L. kunkelii. AB - We examined data for 11 allozyme loci in 14 populations that represent the distribution of the endangered Lotus kunkelii, the narrowly distributed L. arinagensis (both endemic to Gran Canaria), and the broad-ranging L. lancerottensis (endemic to the easternmost Canary Islands, Fuerteventura and Lanzarote) to explore and construe patterns of genetic variation and use this data to assess the controversial taxonomic status of L. kunkelii relative to L. lancerottensis. While L. kunkelii maintains low levels of variation, presumably as a consequence of prolonged inbreeding due to very low population size and sharp geographic isolation, the other two taxa have much higher indicators of polymorphism than those reported for other oceanic island endemics. Lotus arinagensis has the highest genetic polymorphism and the lowest interpopulation differentiation, presumably because of its considerable antiquity and habitat stability, despite recent fragmentation. The high interpopulation differentiation in L. lancerottensis is attributed to the Atlantic acting as a barrier, reducing gene flow within islands. Evolutionary analysis of the allozyme evidence indicates that L. kunkelii is genetically closer to L. arinagensis than to L. lancerottensis, thereby dispelling the taxonomic uncertainty and supporting L. kunkelii as a distinct species, warranting legal protection in the forthcoming catalog of threatened Canarian species. PMID- 21642178 TI - Sources of variation in growth, form, and survival in dwarf and normal-stature pitch pines (Pinus rigida, Pinaceae) in long-term transplant experiments. AB - Determining the relative contributions of genetic and environmental factors to phenotypic variation is critical for understanding the evolutionary ecology of plant species, but few studies have examined the sources of phenotypic differentiation between nearby populations of woody plants. We conducted reciprocal transplant experiments to examine sources of variation in growth rate, form, survival, and maturation in a globally rare dwarf population of pitch pine (Pinus rigida) and in surrounding populations of normal-stature pitch pines on Long Island, New York. Transplants were monitored over a 6-yr period. The influence of seedling origin on height, growth rate, survival, and form (single stemmed vs. multi-stemmed growth habit) was much smaller than the effect of transplanting location. Both planting site and seed origin were important factors in determining time to reproduction; seedlings originating from dwarf populations and seedlings planted at the normal-stature site reproduced earliest. These results suggest that many of the differences between dwarf and normal-stature pitch pines may be due more to plastic responses to environmental factors than to genetic differentiation among populations. Therefore, preservation of the dwarf pine habitat is essential for preserving dwarf pine communities; the dwarf pines cannot be preserved ex situ. PMID- 21642179 TI - Irregular droughts trigger mass flowering in aseasonal tropical forests in asia. AB - General flowering is a community-wide masting phenomenon, which is thus far documented only in aseasonal tropical forests in Asia. Although the canopy and emergent layers of forests in this region are dominated by species of a single family, Dipterocarpaceae, general flowering involves various plant groups. Studying proximate factors and estimating the flowering patterns of the past and future may aid our understanding of the ecological significance and evolutionary factors behind this phenomenon. Here we show that this phenomenon is most likely triggered by irregular droughts based on 10 years of observations. In the aseasonal forests of SE Asia, droughts tend to occur during transition periods from La Nina to El Nino, which results in an irregular 6-7-yr cycle involving a dry period with several droughts and a wet period without droughts. The magnitude of a flowering event also depends on the timing of droughts associated with the El Nino southern oscillation (ENSO) cycle, with the largest events occurring after an interval of several years with no flowering. Because most plant species can only reproduce successfully during large flowering events, changes in the ENSO cycle resulting from global warming, may have serious ramifications for forest regeneration in this region. PMID- 21642180 TI - Phylogenetics of the florally diverse Andean clade Iochrominae (Solanaceae). AB - Recent molecular phylogenetic studies of Solanaceae have identified many well supported clades within the family and have permitted the creation of a phylogenetic system of classification. Here we estimate the phylogeny for Iochrominae, a clade of Physaleae sensu Olmstead et al. (1999), which contains 34 Andean species encompassing an immense diversity of floral forms and colors. Using three nuclear regions, ITS, the second intron of LEAFY, and exons 2 to 9 of the granule-bound starch synthase gene (waxy), we evaluated the monophyly of the traditional genera comprising Iochrominae and assessed the extent of interspecific hybridization within the clade. Only one of the six traditionally recognized genera of Iochrominae was supported as monophyletic. Further, comparison of the individual nuclear data sets revealed two interspecific hybrid taxa and a third possible case. These hybrid taxa occur in the Amotape Huancabamba zone, a region between the northern and central Andes that has the greatest diversity of Iochroma species and offers frequent opportunities for hybridization in areas of sympatry. We postulate that periodic hybridization events in this area coupled with pollinator-mediated selection and the potential for microallopatry may have acted together to promote diversification in montane Andean taxa, such as Iochrominae. PMID- 21642181 TI - The utility of automated analysis of inter-simple sequence repeat (ISSR) loci for resolving relationships in the Canary Island species of Tolpis (Asteraceae). AB - Plants of oceanic islands, often remarkably divergent morphologically from continental relatives, are useful models for studying evolution and speciation because evolution is telescoped in time and space. Prior studies revealed little DNA sequence variation within the clade of ca. 10 Canary Island species of Tolpis, which precluded resolving species relationships. The present study assessed the utility of automated analysis of inter-simple sequence repeat (ISSR) loci for resolving relationships within the clade using 264 individuals from 36 populations of all recognized species and three undescribed morphological variants. Similarity (Dice coefficient) and Fitch parsimony were used to generate neighbor-joining (NJ) and strict consensus trees (MP), respectively. All individuals of the morphologically distinct endemic species formed clusters in both trees. There is also support for clusters of two undescribed variants in the NJ tree. Individuals from a morphologically variable complex consisting primarily of two species are not well resolved at population or species levels. The NJ and MP trees are not congruent at deeper levels, including relationships among species. Results are interpreted in terms of the biology of the species, and the utility of automated analysis of ISSR markers for interpreting patterns of evolution of Tolpis in the Canary Islands is discussed. PMID- 21642182 TI - Patterns of long-distance dispersal in Tiquilia subg. Tiquilia (Boraginaceae): implications for the origins of amphitropical disjuncts and Galapagos Islands endemics. AB - Plant biogeographers have long argued whether plant disjunctions result from vicariance or dispersal. One of the classic patterns of plant disjunction involves New World amphitropical disjuncts, as exemplified by Tiquilia subg. Tiquilia (Boraginaceae). Subgenus Tiquilia forms a heterogeneous group of ~20 species that is amphitropically distributed in the deserts of North and South America, with four taxa endemic to the Galapagos Islands. The current study reconstructs the biogeographic history of subg. Tiquilia in order to explore the origins of New World amphitropical disjunction and of Galapagos endemism. A strongly supported phylogeny of the subgenus is estimated using sequence data from matK, ndhF, rps16, ITS, and waxy. Biogeographic analyses using combined and individual marker data sets reveal a complex history of long-distance dispersal in subg. Tiquilia. Biogeographic reconstructions imply a North American origin of the subgenus and its three major lineages and require at least four long-distance dispersal events to explain its current distribution. The South American taxa of subg. Tiquilia result from three independent and nonsimultaneous colonization events, while the monophyly and continental origins of the Galapagos endemics are unresolved. This study contributes to a growing body of evidence that intercontinental dispersal has been more common than previously realized. PMID- 21642183 TI - Infrageneric phylogeny of Schoenocaulon (Liliales: Melanthiaceae) with clarification of cryptic species based on ITS sequence data and geographical distribution. AB - As currently defined, the 24 species of Schoenocaulon occur in three disjunct areas: north central Florida (one species, S. dubium), southern Peru (portion of the range of S. officinale), and the region from southeastern New Mexico-Texas south to Venezuela; the 20 species endemic to Mexico are geographically restricted. Species delimitations, often based on tepal morphology, have been problematic. Our analyses of ITS sequence data for all 27 species and infraspecific taxa support recognition of two new species and recircumscription and placement of elements of the polyphyletic S. ghiesbreghtii and S. mortonii complexes. For taxa with adequate sampling, our data also indicate 11-12 cladospecies and 3-6 metaspecies according to the apomorphic species concept. The resolved phylogeny, correlated with geography and morphology, allows insight into biogeographical diversification and the evolution of some unusual morphological characters within the genus, such as nectary differentiation and tepal margin type. PMID- 21642184 TI - Molecular phylogenetics reveals Leontodon (Asteraceae, Lactuceae) to be diphyletic. AB - The plastid matK gene, trnL/F spacer, and nuclear rDNA ITS were sequenced for 36 species of Leontodon and 29 taxa of related genera of tribe Lactuceae. Phylogenetic relationships inferred from the independent and combined data are largely congruent and reveal that Leontodon sensu lato (s.l.) as presently defined is diphyletic: L. subgenus Leontodon forms a clade with Helminthotheca, Picris and Hypochaeris as sister genera, whereas L. subgenus Oporinia appears as a separate clade with strong bootstrap support and is thus better treated as a separate genus. Previous sectional classifications of Leontodon s.l. are considered in the light of DNA and additional morphological and karyological data. Support is presented for a core group of Hypochaeridinae sensu stricto (s.s.) with the two clades of Leontodon s.l., Helminthotheca, Picris, and Hypochaeris, whereas Urospermum, Hyoseris, Aposeris, and Rhagadiolus appear to be positioned more distantly. PMID- 21642185 TI - Worldwide phylogeny and biogeography of Cardamine flexuosa (Brassicaceae) and its relatives. AB - Phylogenetic relationships, biogeography, and taxonomy of a group of taxa putatively related to the tetraploid Cardamine flexuosa were explored using sequences of the internal transcribed spacer region of nrDNA (ITS) and the trnL trnF region of cpDNA. Taxon sampling focused on eastern Asia, North America, and Europe, and included 19 taxa represented by 177 and 182 accessions for each data set, respectively. Our analyses provided unequivocal evidence that Asian weedy populations traditionally assigned to C. flexuosa form an independent evolutionary lineage and represent a distinct taxon from European C. flexuosa. The allopolyploid origin of this common weed in paddy fields, its origin, and/or spread associated with the establishment of suitable man-made habitats are suggested. It is also found as an introduced weed in Australia and North America. Phylogenetic relationships and the associated taxonomic implications are presented and discussed for the group as a whole. Contrasting patterns of genetic variation (particularly in cpDNA) among different species were revealed. While very little haplotype diversity was found in widespread C. hirsuta and C. flexuosa, greater variation, showing phylogeographic structure, was observed in the tetraploid C. scutata within a relatively small area of Japan. PMID- 21642187 TI - Erratum. PMID- 21642186 TI - Phylogenetic relationships and generic delimitation in subtribe Arctotidinae (Asteraceae: Arctotideae) inferred by DNA sequence data from ITS and five chloroplast regions. AB - Asteraceae are the largest family in southern Africa. Elucidating its origins and radiation in the region requires well-supported species-level phylogenies of the lineages. This paper presents a phylogenetic framework for subtribe Arctotidinae, which have a southern and eastern African-Australian distribution centered in the winter-rainfall region of South Africa. DNA sequence data from five chloroplast fragments (ndhF, psbA-trnH, rps16, trnS-trnfM, and trnT-trnF) and the nuclear ITS region were analyzed separately and in combination using parsimony and Bayesian methods. The data sets comprised exemplars from 18 ingroup species, representing the five currently accepted genera, and four outgroup species from Gorteriinae. All analyses indicated Arctotis and Haplocarpha are polyphyletic as presently circumscribed. The Australian-endemic Cymbonotus lawsonianus was placed within a strongly supported clade also containing A. arctotoides from South Africa and H. schimperi from eastern Africa. Retention of Dymondia and resurrection of Landtia at generic level are strongly supported. The phylogenetic hypotheses indicate the subtribe might have originated in temperate southern or eastern Africa, or it was ancestrally widespread in southern Africa and has diversified vicariously. The derived placement of C. lawsonianus indicates long-distance dispersal from southern Africa to Australia occurred. PMID- 21642188 TI - Flower or spikelet? Understanding the morphology and development of reproductive structures in Exocarya (Cyperaceae, Mapanioideae, Chrysitricheae). AB - Fundamental questions of floral morphology remain unresolved in the grasslike monocots in order Poales, including what constitutes a flower and what constitutes a spikelet. The mapaniid sedges have particularly complex spikeletlike structures, variously interpreted as clusters of flowers or spikelets. Recent phylogenetic studies of Cyperaceae have identified the mapaniid clade as sister to the rest of the family, but the homology of mapaniid reproductive units (RUs) and spikeletlike units (SLUs) to other sedge flowers and spikelets is unclear. We examined reproductive development in the mapaniid Exocarya sclerioides. Inflorescence branches terminated in a SLU with bracts and 1-4 RUs. RUs had four small leaflike structures (LLSs): two lateral LLSs, each associated with a stamen, an abaxial LLS associated with a stamen, and an adaxial LLS. The gynoecium terminated the RU. All RUs were axillary to bracts, and unexpanded bracts and RUs were produced beyond expanded RUs, so SLUs were racemose. RUs developed from a single primordium that initiated two lateral LLSs, then two lateral stamens, then the gynoecium. Initiation of the abaxial LLS and stamen and the adaxial LLS followed. We hypothesize that the RU is a sympodial branch that terminates in a hermaphroditic flower with two stamens and a gynoecium; the two lateral LLSs are halves of a deeply divided prophyll. PMID- 21642189 TI - A biomechanical perspective on the role of large stem volume and high water content in baobab trees (Adansonia spp.; Bombacaceae). AB - The stems of large trees serve in transport, storage, and support; however, the degree to which these roles are reflected in their morphology is not always apparent. The large, water-filled stems of baobab trees (Adansonia spp.) are generally assumed to serve a water storage function, yet recent studies indicate limited use of stored water. Through an analysis of wood structure and composition, we examined whether baobab morphology reflects biomechanical constraints rather than water storage capacity in the six Madagascar baobab species. Baobab wood has a high water content (up to 79%), low wood density (0.09 0.17 g . cm(-3)), high parenchyma content (69-88%), and living cells beyond 35 cm into the xylem from the cambium. Volumetric construction cost of the wood is several times lower than in more typical trees, and the elastic modulus approaches that of parenchyma tissue. Safety factors calculated from estimated elastic buckling heights were low, indicating that baobabs are not more overbuilt than other temperate and tropical trees, yet the energy investment in stem material is comparable to that in temperate deciduous trees. Furthermore, the elastic modulus of the wood decreases with water content, such that excessive water withdrawal from the stem could affect mechanical stability. PMID- 21642190 TI - Inter-tracheid pitting and the hydraulic efficiency of conifer wood: the role of tracheid allometry and cavitation protection. AB - Plant xylem must balance efficient delivery of water to the canopy against protection from air entry into the conduits via air-seeding. We investigated the relationship between tracheid allometry, end wall pitting, safety from air seeding, and the hydraulic efficiency of conifer wood in order to better understand the trade-offs between effective transport and protection against air entry. Root and stem wood were sampled from conifers belonging to the Pinaceae, Cupressaceae, Podocarpaceae, and Araucariaceae. Hydraulic resistivity of tracheids decreased with increasing tracheid diameter and width, with 64 +/- 4% residing in the end wall pitting regardless of tracheid size or phylogenetic affinity. This end-wall percentage was consistent with a near-optimal scaling between tracheid diameter and length that minimized flow resistance for a given tracheid length. There was no evidence that tracheid size and hydraulic efficiency were constrained by the role of the pits in protecting against cavitation by air-seeding. An increase in pit area resistance with safety from cavitation was observed only for species of the northern hemisphere (Pinaceae and Cupressaceae), but this variable was independent of tracheid size, and the increase in pit resistance did not significantly influence tracheid resistance. In contrast to recent work on angiosperm vessels, protection against air-seeding in conifer tracheids appears to be uncoupled from conduit size and conducting efficiency. PMID- 21642191 TI - Genetic diversity and population structure in natural populations of Moroccan Atlas cedar (Cedrus atlantica; Pinaceae) determined with cpSSR markers. AB - Atlas cedar (Cedrus atlantica) is an ecologically and economically important forest tree species of northern Africa and is considered one of the endangered conifer species in the region. Chloroplast microsatellites (cpSSR) were used to study genetic variation within and among populations and geographical structure in natural populations of C. atlantica throughout its entire distribution range in Morocco. A total of 25 chloroplast haplotypes and 66 cpSSR alleles were found among 162 individuals. The cpSSRs indicate that C. atlantica appears to maintain a high level of genetic diversity (mean H(e) = 0.95), as observed in most coniferous species. Values of mean pairwise distance within a population (D(2)(SH)) were related to the size and location of the populations. AMOVA analysis showed that most of the variation in C. atlantica occurs within populations and confirmed the general tendency of gymnosperms to display lower values of population differentiation than angiosperms. The distance-based clustering method (PCoA and neighbor-joining analysis) and the geographical structure revealed a poor structure among the six populations of Cedrus atlantica. Also, a Mantel test indicated a weak correlation between geographic and genetic distances (P = 0.106, r = 0.363). These results are also interpreted in the context of postglacial history of the region plus human impacts. PMID- 21642192 TI - Fine-scale genetic structure of the common Primula elatior (Primulaceae) at an early stage of population fragmentation. AB - Many rare species are threatened by habitat fragmentation; however, less is known about effects of fragmentation on common species, despite their potential role in ecosystem productivity and functioning. We identified key factors and processes influencing gene flow in a large population of Primula elatior, a common distylous perennial herb, at an early stage of the fragmentation process, i.e., when fragmentation is taking place. Using 19 allozyme loci, we investigated genetic variation and fine-scale spatial genetic structure (SGS) at seedling and adult life stages in relation to fragmentation history (recent bottlenecks), selection, clonal propagation, sexual reproduction (seed and pollen dispersal, distyly), and patchy structure (patch size, plant density, and morph ratio). The main factors contributing to the strong SGS are seed and (to a lesser extent) pollen dispersal, through a spatial Wahlund effect and biparental inbreeding. Significant differences in allele frequencies between seedlings and adults indicate a temporal Wahlund effect. Patch plant density and biased morph ratio also affect the genetic patterns. Our results show that if P. elatior populations evolve into patchworks of small, isolated remnants, genetic erosion, reduced gene flow, and increased inbreeding can be expected, suggesting that such common plant species might require large population sizes to remain viable. PMID- 21642193 TI - Reproductive biology of three fern species may contribute to differential colonization success in post-agricultural forests. AB - Because selfing enables a single individual to reproduce in a new location, the ability to self-fertilize should enhance plants' capacity for colonization. This study examined whether selfing ability correlated with successful migration in three fern species, Dryopteris carthusiana, Dryopteris intermedia, and Polystichum acrostichoides, which vary in their ability to colonize forests on abandoned agricultural lands in central New York, USA. Polystichum acrostichoides is much more frequent in forests that were never cleared for agriculture, D. carthusiana is more frequent in forests that developed on former fields, and D. intermedia is equally frequent in the two forest types. To test the hypothesis that better-colonizing species and post-agricultural forest populations have greater selfing ability, I assessed the sporophyte production of gametophytes grown in isolation and in pairs of varying relatedness. Dryopteris carthusiana had the highest reproductive success and selfing ability and P. acrostichoides the lowest. These results support the hypothesis that selfing may facilitate colonization in these species. They also exemplify the general pattern that polyploid fern species have higher rates of self-fertilization than related diploids, as the allotetraploid D. carthusiana had greater selfing ability than both diploid species. PMID- 21642194 TI - Reproductive biology of the dioecious Canary Islands endemic Withania aristata (Solanaceae). AB - We confirmed functional dioecy of Withania aristata via field and greenhouse studies. Male flowers are significantly larger. Female flowers bear stamens with no pollen; males bear 220 000 grains. Stigmata of male flowers senesce in buds. Anatomical observations confirm more ovules in females and an ovarian nectary in both sexes. We detected nectar in female flowers in the greenhouse but found no nectar in males. Thus, males offer pollen and females nectar. Females bear large numbers of fruits and, infrequently, male plants bear few significantly smaller fruits with few seeds. Outcrosses of females (self crosses impossible without pollen) yielded fruits in young buds, older buds, and open flowers. Self crosses of male flowers succeeded only with very young buds. Although functionally dioecious, this species manifests self-compatibility; however, no fruits are produced autonomously. Bee species (Lassioglossum, Amegilla, Apis) visit flowers and mature buds. Bud visits in which bees force petal tips apart, coupled with self-compatibility, may explain infrequent fruit on males. Thus, dioecy in W. aristata seems to have evolved from self-compatible ancestors, that leaky dioecy may have been favored during colonization, and, that despite autogamy and a low floral visition rate, this endemic enjoys a high rate of reproductive success. PMID- 21642195 TI - Multiple pollinator visits to Mimulus ringens (Phrymaceae) flowers increase mate number and seed set within fruits. AB - The timing and effectiveness of pollinator visitation to flowers is an important factor influencing mating patterns and reproductive success. Multiple pollinator probes to a flower may increase both the quantity and genetic diversity of progeny, especially if single probes deposit insufficient pollen for maximal seed set or if the interval between probes is brief. When pollen carryover is limited, sequential pollen loads may also differ markedly in sire representation. We hypothesized that these conditions help explain high levels of multiple paternity in Mimulus ringens fruits. We documented all bee visits to individual flowers, quantified resulting seed set, and determined paternity for 20 seeds per fruit. Most (76%) flowers received multiple probes, and the interval between probes was usually <30 min. Flowers probed multiple times produced 44% more seeds than flowers probed once. All fruits were multiply sired. Flowers receiving a single probe averaged 3.12 outcross sires per fruit, indicating that single probes deposit pollen from several donors. Multiple paternity was even greater after three or more probes (4.92 outcross sires), demonstrating that sequential visits bring pollen from donors not represented in the initial probe. PMID- 21642196 TI - Reproductive effort and costs of reproduction do not explain female-biased sex ratios in the moss Pseudocalliergon trifarium (Amblystegiaceae). AB - A fundamental assumption in life-history theory is that reproduction is costly. Higher reproductive investment for fruits than for flowers may result in larger costs of reproduction in females than in males, which is often used to explain male-skewed sex ratios in unisexual seed plants. In contrast, bryophytes have predominantly female-biased sex ratios, suggested to be a product of a higher average cost of sexual reproduction in males. Empirical evidence to support this notion is largely lacking. We investigated sex-specific reproductive effort and costs in the unisexual moss Pseudocalliergon trifarium that has a female dominated expressed sex ratio and rarely produces sporophytes. Annual vegetative segment mass did not differ among male, female, and non-expressing individuals, indicating that there was no threshold-size for sex expression. Mean and annual mass of sexual branches were higher in females than in males, but branch number per segment did not differ between sexes. Prefertilization reproductive effort for females was significantly greater (11.2%) than for males (8.6%). No cost for sexual branch production in terms of reduced relative vegetative growth or decreased investment in reproductive structures in consecutive years was detected. A higher realized reproductive cost in males cannot explain the unbalanced sex ratio in the study species. PMID- 21642197 TI - Relationship between floral longevity and sex allocation among flowers within inflorescences in Aquilegia buergeriana var. oxysepala (Ranunculaceae). AB - Understanding the fitness of plants with inflorescences requires examining variation in sex allocation among flowers within inflorescences. We examined whether differences in the duration of the male and female phases of flowering lead to variation in sex allocation and reproductive success among flowers within inflorescences. In 2002 and 2003, we quantified floral longevity, floral sex allocation, and reproductive success between the first and the second flowers within inflorescences in a protandrous species, Aquilegia buergeriana var. oxysepala. Floral longevity was greater in the first flowers than in the second ones in both years. The male phase lasted longer, and the initial number of pollen grains and the number of pollen grains removed were greater in the first flowers than in the second ones in both years. Within first flowers, the number of pollen grains removed was greater in flowers that had longer male phases, thus duration of the male phase may positively affect male reproductive success in the first flowers. The female phase lasted longer and the number of ovules was greater in the first flowers than in the second only in 2002. However, seed production per flower and female phase duration in both years were not significantly related. The variation in the number of pollen grains among flowers in this species may be caused by the variation in male phase duration. PMID- 21642198 TI - Molecular phylogeny and dating reveals an Oligo-Miocene radiation of dry-adapted shrubs (former Tremandraceae) from rainforest tree progenitors (Elaeocarpaceae) in Australia. AB - To better understand the historical biogeography of the southern hemisphere and evolutionary responses of plants to aridity, we undertook a detailed phylogenetic study of the predominantly southern family Elaeocarpaceae sensu lato (including Tremandraceae). Plastid trnL-trnF and nuclear ITS sequence data were analyzed using parsimony and Bayesian methods and molecular evolutionary rates calibrated using the Oligocene fossil record of Elaeocarpus mesocarps to estimate the minimum divergence dates. The results indicate the monophyly of all recognized genera and a placement for the former Tremandraceae (three genera and about 49 species of shrubby, dry-adapted Australian plants) within the widespread predominantly rainforest tree family Elaeocarpaceae (nine genera, over 500 species). The former Tremandraceae clade diverged from its sister (Aceratium + Elaeocarpus + Sericolea) during the Paleocene, after which it underwent a marked acceleration in evolutionary rate. Furthermore, this lineage diversified during the late Miocene, coincident with widespread aridification in Australian environments and extensive radiations of several sclerophyllous groups. The role of dispersal in explaining the current geographical distribution of Elaeocarpaceae is illustrated by Aristotelia. This genus, whose distribution was previously thought to reflect Gondwanan vicariance, is shown to have arrived in New Zealand from Australia at least 6-7 million yr ago. PMID- 21642199 TI - Evolution of biogeographic disjunction between eastern Asia and eastern North America in Phryma (Phrymaceae). AB - This study examines molecular and morphological differentiation in Phryma L., which has only one species with a well-known classic intercontinental disjunct distribution between eastern Asia (EA) and eastern North America (ENA). Phylogenetic analysis of nuclear ribosomal ITS and chloroplast rps16 and trnL-F sequences revealed two highly distinct clades corresponding to EA and ENA. The divergence time between the intercontinental populations was estimated to be 3.68 +/- 2.25 to 5.23 +/- 1.37 million years ago (mya) based on combined chloroplast data using Bayesian and penalized likelihood methods. Phylogeographic and dispersal-vicariance (DIVA) analysis suggest a North American origin of Phryma and its migration into EA via the Bering land bridge. Multivariate analysis based on 23 quantitative morphological characters detected no geographic groups at the intercontinental level. The intercontinental populations of Phryma thus show distinct molecular divergence with little morphological differentiation. The discordance of the molecular and morphological patterns may be explained by morphological stasis due to ecological similarity in both continents. The divergence of Phryma from its close relatives in the Phrymaceae was estimated to be at least 32.32 +/- 4.46 to 49.35 +/- 3.18 mya. PMID- 21642200 TI - The taxonomic value of fruit wing types in the order Apiales. AB - This study shows that structural data, when carefully examined, can provide valuable characters for delimiting monophyletic groups and can complement DNA with observable features to recognize and circumscribe taxa. In the angiosperm order Apiales, traditional classification has relied heavily (often exclusively) on fruit characters. Recent molecular systematic studies, however, provided a radically different picture of relationships, calling into question the utility of fruit characters. We have studied fruit anatomy from 18 genera (Annesorhiza, Asteriscium, Astrotricha, Choritaenia, Dasispermum, Elaeoselinum, Heptaptera, Hermas, Heteromorpha, Laretia, Molopospermum, Myodocarpus, Pachypleurum, Peucedanum, Polemanniopsis, Polylophium, Rouya, and Tordylium) that represent all major taxonomic groups of Apiales characterized by winged fruits and the full range of wing types. Fruit anatomy closely corresponded with the phylogenetic position of these genera, as suggested by molecular studies. Fruit features of taxonomic importance include developmental origin of the wings, carpel shape, presence of vittae, woodiness of the endocarp, position of crystals, and type of carpophores. Despite the long history of recognizing umbellifers as a "natural group," few studies have been able to provide structural characters to help circumscribe the clades identified by molecular data. The interpretations presented are an important step toward erecting a stable system of classification for this difficult family. PMID- 21642201 TI - Erratum. PMID- 21642202 TI - Stem diversity, cauline domatia, and the evolution of ant-plant associations in Piper sect. Macrostachys (Piperaceae). AB - Plants possess a variety of structures that harbor ant nests, and the morphology of these domatia determines the nature of ant-plant mutualisms in a given plant species. In this study, we report on the differences in anatomy between myrmecophytes of Piper, which are regularly excavated by an obligate ant mutualist (Pheidole bicornis) and nonmyrmecophytes of Piper, which consistently have solid stems. Stems of excavated plant species lacked outward evidence of modification; however, striking anatomical differences were apparent between hollow-stemmed species before excavation and the remainder of the solid-stemmed species studied. Prior to excavation by ants, stems of myrmecophytes were characterized by strongly heterogeneous piths in which a large, central area had relatively large cells lacking intracellular crystals with a periphery of smaller cells containing numerous crystals. The domatium excavated by the ants was restricted to the large-celled region. This is the first report of the absence of crystals in ant-excavated portions of stems of myrmecophytes. Cauline domatia became lined with 3-8 cell layers of suberized wound tissue, which may have an impact on nutrient absorption by Piper myrmecophytes. PMID- 21642203 TI - Amyloplast to chromoplast conversion in developing ornamental tobacco floral nectaries provides sugar for nectar and antioxidants for protection. AB - Tobacco floral nectaries undergo changes in form and function. As nectaries change from green to orange, a new pigment is expressed. Analysis demonstrated that it is beta-carotene. Plastids undergo dramatic changes. Early in nectary development, they divide and by stage 9 (S9) they are engorged with starch. About S9, nectaries shift from quiescent anabolism to active catabolism resulting in starch breakdown and production of nectar sugars. Starch is replaced by osmiophilic bodies, which contain needle-like carotenoid crystals. Between S9 and S12, amyloplasts are converted to chromoplasts. Changes in carotenoids and ascorbate were assayed and are expressed at low levels early in development; however, following S9 metabolic shift, syntheses of beta-carotene and ascorbate greatly increase in advance of expression of nectar redox cycle. Transcript analysis for carotenoid and ascorbate biosynthetic pathways showed that these genes are significantly expressed at S6, prior to the S9 metabolic shift. Thus, formation of antioxidants beta-carotene and ascorbate after the metabolic shift is independent of transcriptional regulation. We propose that biosynthesis of these antioxidants is governed by availability of substrate molecules that arise from starch breakdown. These processes and events may be amenable to molecular manipulation to provide a better system for insect attraction, cross pollination, and hybridization. PMID- 21642204 TI - Old fronds serve as a vernal carbon source in the wintergreen fern Dryopteris intermedia (Aspleniaceae). AB - Maintaining green leaves beyond the growing season has been hypothesized to benefit plants by supplying either a nutrient or a carbon source. Understanding such ecophysiological aspects of plants will help us to appreciate how a species functions in its environment and predict how it might be affected by future changes in that environment. The wintergreen fern species Dryopteris intermedia does not retranslocate nitrogen and phosphorus from old fronds in spring, but photosynthesis does take place in the old fronds during this season. To determine if carbon fixed in the old fronds is translocated to other parts of the plant, we labeled old fronds with (13)C via photosynthetic uptake and examined old fronds, new fronds, fine roots, and rhizomes for (13)C content 1 day and 1 month after labeling the old fronds. Vernally fixed carbon was translocated to the new fronds but not significantly to the below ground tissues. Old fronds in this species, therefore, serve as a carbon source for vernal growth of new fronds. This is the first study in which a fern was labeled with (13)C to track vernally fixed carbon from old fronds to the rest of the plant in a wintergreen species. Future research should examine the precise timing of this carbon movement and examine other species for a similar or contrasting strategy. PMID- 21642205 TI - On the origin of the sweet-smelling Parma violet cultivars (Violaceae): wide intraspecific hybridization, sterility, and sexual reproduction. AB - Parma violets are reputed for their double, fragrant flowers and have been cultivated for centuries in Europe. However, due to a rather atypical morphology their taxonomic affinity has not been clarified. Authors have proposed an origin from three possible species, Viola alba, V. odorata, or V. suavis, or a hybrid origin. Using both ITS sequence variation and allozyme variation in 14 putative loci, we showed that the Parma violet cultivars have their origin within Viola alba and that they are best included in the Mediterranean subsp. dehnhardtii. There is no trace of interspecific hybridization. However, the cultivars appear to have a single origin in a wide hybrid within V. alba, involving parental plants from the eastern and western Mediterranean region; historical literature sources seem to indicate Turkey and Italy, respectively. The Parma violet cultivars possess high levels of allozyme heterozygosity and to some extent also within-individual ITS sequence variation. Losses of heterozygosity and within individual ITS sequence variation in some of the cultivars indicate subsequent rare events of sexual reproduction, presumably through cleistogamous seed set. We unambiguously identify the closest wild relative of this group of cultivars, allowing growers to develop new selection procedures, and show a peculiar molecular process associated with human selection. PMID- 21642206 TI - Paternal, maternal, and biparental inheritance of the chloroplast genome in Passiflora (Passifloraceae): implications for phylogenetic studies. AB - Patterns of inheritance of the chloroplast genome in Passiflora were analyzed by examining the progeny from both interspecific and intraspecific crosses. Artificial crosses of field-collected material were performed in greenhouses at The University of Texas at Austin. DNA from fresh leaf material was analyzed by Southern blot techniques to identify the donor of the chloroplast genome. Initially, single progeny were analyzed for 11 crosses; two intraspecific crosses demonstrated maternal inheritance, whereas the nine interspecific crosses had paternal inheritance. Subsequently, the donor of the chloroplast genome was determined for multiple progeny in seven crosses. Passiflora oerstedii * P. retipetala showed strict paternal inheritance in all of 17 progeny. A series of five crosses and backcrosses between P. oerstedii and P. menispermifolia demonstrated strictly paternal inheritance. Finally, when 15 progeny were analyzed for the P. costaricensis * P. costaricensis cross, 12 of the 15 showed maternal inheritance, whereas the remaining three were biparental. Interestingly, all interspecific crosses had primarily paternal inheritance, whereas all intraspecific crosses had primarily maternal inheritance. The implications of heteroplasmy on phylogenetic analyses of chloroplast DNA are discussed. PMID- 21642207 TI - Specialization for pollination by beetles and wasps: the role of lollipop hairs and fragrance in Satyrium microrrhynchum (Orchidaceae). AB - Exposed nectar presentation is a key trait in flowers specialized for pollination by short-tongued insects. We investigated the pollination of Satyrium microrrhynchum, a rare South African orchid in which nectar is secreted as droplets on long floral hairs ("lollipop hairs") at the mouth of a shallow labellum. Our observations indicate that this orchid is pollinated specifically by two insect species: a cetoniid beetle (Atrichelaphinus tigrina) and a pompilid wasp (Hemipepsis hilaris). Both insects have short mouthparts and remove nectar from the hairs with sweeping motions of their mouthparts. Pollinaria become attached to the upper surface of their heads while they feed on the nectar. Beetles damage the hairs while feeding, which may explain the positive relationship between hair damage and pollination success in plants of S. microrrhynchum from populations where beetles were common. The orchid has cryptic green-yellow flowers with spectral reflectance similar to that of its leaves. The fragrance from plants in three populations, analyzed using gas chromatography coupled to mass spectrometry, was dominated by various terpenoids; linalool was the most abundant. Plants in different populations emitted similar compounds, but eugenol and derivatives of this compound were found in only one of the three populations. In an electrophysiological study (gas chromatography coupled to electroantennography), using antennae of A. tigrina, clear signals were elicited by some of the floral scent compounds. PMID- 21642208 TI - Early ontogenetic patterns in chemical defense in Plantago (Plantaginaceae): genetic variation and trade-offs. AB - Predictions based on the plant age and growth-differentiation balance hypotheses of defense were tested in two congeneric species, Plantago lanceolata and P. major, by quantifying iridoid glycosides, defensive chemicals, in seeds and leaves during the first 6 wk of growth. Concentrations decreased from the seed to 2-wk-old seedling stage in P. lanceolata, but increased during this period in P. major. In both species, levels were similar for 2- and 4-wk-old plants, then significantly increased from 4 to 6 wk. Genetic variation in the ontogeny of iridoid glycoside production was significant in both species at the maternal family level and at the population level. To examine whether allocation costs could explain the low production of iridoid glycosides in seedlings, relationships between growth and defense (iridoid glycosides) were characterized. Growth and defense had a positive or null relationship in all age groups, indicating that there was no trade-off in these plants at any age. This study provides some support for the growth-differentiation balance hypothesis, but offers no support for the plant age hypothesis. Measuring how herbivory affects plant fitness at different ontogenetic stages may shed light on these patterns in Plantago and on the evolution of the ontogeny of defense. PMID- 21642209 TI - Determinants of biased sex ratios and inter-sex costs of reproduction in dioecious tropical forest trees. AB - Estimates of the sex ratio and cost of reproduction in plant populations have implications for resource use by animals, reserve design, and mechanisms of species coexistence, but may be biased unless all potentially reproductive individuals are censused over several flowering seasons. To investigate mechanisms maintaining dioecy in tropical forest trees, we recorded the flowering activity, sexual expression, and reproductive effort of all 2209 potentially reproductive individuals within 16 species of Myristicaceae over 4 years on a large forest plot in Amazonian Ecuador. Female trees invested >10 times more biomass than males in total reproduction. Flowering sex ratios were male-biased in four species in >=1 year, and cumulative 4-year sex ratios were male-biased in two species and for the whole family, but different mechanisms were responsible for this in different species. Annual growth rates were equivalent for both sexes, implying that females can compensate for their greater reproductive investment. There was no strict spatial segregation of the sexes, but females were more often associated with specific habitats than males. We conclude that male-biased sex ratios are not manifested uniformly even after exhaustive sampling and that the mechanisms balancing the higher cost of female reproduction are extremely variable. PMID- 21642210 TI - Species richness along a production gradient: a multivariate approach. AB - The specific shape of the relationship between plant diversity and productivity and the causal mechanism(s) behind the observed pattern(s) are still highly debated. Recent advances suggest that the relationship depends on several environmental variables and may change with the observational scale. In this study, a multivariate, multiscale approach was used to identify the variables that determine the relationship between species richness and annual production along a forest/old field edge in southern Quebec (Canada). Various relationships between richness and production were found at different distances to the edge. In the forest, most relationships were positive and linear, while in the old field the relationship shifted from positive linear to non-significant with increasing distance from the edge. In the forest or in the old field, the shape of the relationship (all distances from the edge combined) was unimodal. Path analyses showed that species richness was determined mostly by production, which was influenced by different limiting resources, depending on the community (forest or old field). An increasing range in production created by pooling across community types can confound the resources and/or conditions determining the diversity productivity relationship. PMID- 21642211 TI - The closest relatives of cacti: insights from phylogenetic analyses of chloroplast and mitochondrial sequences with special emphasis on relationships in the tribe Anacampseroteae. AB - Recent molecular and morphological systematic investigations revealed that the cacti are most closely related to Anacampseroteae, Portulaca and Talinum of the family Portulacaceae (ACPT clade of suborder Portulacineae). A combined analysis of ndhF, matK, and nad1 sequence data from the chloroplast and the mitochondrial genomes indicates that the tribe Anacampseroteae is the sister group of the family Cactaceae. This clade, together with Portulaca, is well characterized by the presence of axillary hairs or scales. Relationships within Anacampseroteae are characterized by a grade of five species of Grahamia s.l. from North and South America, and Grahamia australiana is found to be sister to the genera Anacampseros and Avonia. A comparison of vegetative characteristics indicates an evolutionary transition from woody subshrubs to dwarf perennial and highly succulent herbs during the diversification of Anacampseroteae. Available evidence from the present investigation as well as from previously published studies suggests that a revised classification of Portulacineae on the basis of inferred phylogenetic relationships might consist of a superfamily that includes Cactaceae and the three genera Anacampseros s.l. (including Avonia and Grahamia s.l.), Portulaca, and Talinum (including Talinella), either referred to three monogeneric families or to a paraphyletic family Portulacaceae*. PMID- 21642212 TI - Agroecosystem management for rare species of Paysonia (Brassicaceae): integrating their seed ecology and life cycle with cropping regimens in a changing climate. AB - Dormancy break and germination of seeds are governed by climatic cues, and predicted changes in climate may impact the ecology and conservation of species. Paysonia perforata and P. stonensis are rare brassicaceous winter annuals occurring primarily in fields on floodplains, where corn or soybeans are recommended for habitat maintenance. We tested the effects of precipitation, based on two predictions of changes in climate, on seed germination in these two species and placed the results into a management framework. Seeds of both species, collected during peak dispersal in late April/early May, were given various periods of light (or darkness) followed by darkness (or light) at summer temperatures before placement in darkness during late summer/early autumn in both laboratory and field. The light requirement was met earliest at 10 wk (mid-July) on alternating wet/dry substrate (simulating current climatic conditions). However, seeds of P. perforata and P. stonensis were photostimulated earliest at 2 wk (mid-May) and 6 wk (mid-June), respectively, on a continuously moist substrate (simulating predicted future conditions). The soil seed bank could be depleted if plowing coincides with photostimulation of seeds. Fields should be prepared after dispersal but before seeds are photostimulated and harvesting completed before seed germination in early September. Because seeds are highly photostimulated in late summer, disturbance from harvesting must be low to prevent burial. Cultivation of soybean, particularly for forage, is better matched to the seed biology and life cycle of Paysonia than that of corn under current and predicted climates. PMID- 21642213 TI - Hierarchical patterns of paternity within crowns of Albizia julibrissin (Fabaceae). AB - The floral architecture and phenology of the tree species Albizia julibrissin (Fabaceae) offer the potential for flowers within inflorescences to share common pollen donors. Patterns of paternity within individual tree crowns may differ among isolated individuals and those in populations due to differences in pollinator foraging behavior. To determine how genetic diversity is partitioned within individual seed pools and whether these patterns differ among isolated and population trees, we obtained all fruits from three inflorescences from four clusters from three isolated trees and from three population trees in Athens, Georgia. We assayed 14 polymorphic allozymes to genotype all progeny within singly sired fruits to determine the multilocus genotype of each fruit's pollen donor. Inflorescences had multiple pollen donors, but simulation analyses revealed that redundancy of pollen donors tended to be more likely within inflorescences than randomly across the crown. Analysis of genetic and genotypic diversity indicated that individual maternal trees received pollen from many donors in uneven frequencies. Results suggest that isolated trees receive pollen from slightly fewer pollen donors and experience more within-plant pollinator movement than trees in populations. However, isolated trees receive qualitatively similar pollen from many sources, suggesting that these trees are not effectively isolated and that pollen moves long distances in this species. PMID- 21642214 TI - Impact of mate availability, population size, and spatial aggregation of morphs on sexual reproduction in a distylous, aquatic plant. AB - In distylous, self-incompatible plants, clonal propagation, unbalanced floral morph frequencies, and reduced population size can interfere with the functioning of distyly by compromising legitimate intermorph pollinations, resulting in reduced reproductive output. Here, we examined the mating system and the impact of mate availability, population size, and spatial aggregation of morphs on reproductive output in the distylous, clonal, aquatic plant Hottonia palustris. Controlled pollinations under greenhouse conditions detected no spontaneous selfing without the action of a pollen vector (autonomous autogamy) and demonstrated very low fruit and seed development after self-pollination. Intermorph (legitimate) crossings resulted in high reproductive output in both floral morphs (long- and short-styled individuals), whereas intramorph (illegitimate) crossings decreased fruit and seed development by more than 50%, indicating that the species has partial intramorph-incompatibility. In natural populations, small population size and increasing deviation of floral morph frequencies negatively affected reproductive outcome. Individuals of the majority morph type developed significantly fewer fruit and seeds than individuals of the minority morph type. This rapid decline in fecundity was symmetrical, indicating that regardless of which morph was in the majority, the same patterns of negative frequency-dependent mating occurred. Increasing spatial isolation between compatible morphs significantly reduced fruit and seed set in both morphs similarly. This study provides clear indications of frequency- and context dependent mating in natural populations of a distylous plant species. PMID- 21642215 TI - Population differentiation and gene flow within a metapopulation of a threatened tree, Magnolia stellata (Magnoliaceae). AB - We examined genetic differentiation among eight local populations of a metapopulation of Magnolia stellata using 10 nuclear and three chloroplast microsatellite (nSSR and cpSSR) markers and evaluated the influence of historical gene flow on population differentiation. The coefficient of genetic differentiation among populations for nSSR (F(ST) = 0.053) was less than half that for cpSSR (0.137). An isolation-by-distance pattern was detected for nSSRs, but not cpSSRs. These results suggest that pollen flow, as well as seed dispersal, has significantly reduced genetic differentiation among populations. We also examined patterns of contemporary pollen flow by paternity analysis of seeds from nine seed parents in one of the populations using the nSSR markers and found it to be greatly restricted by the distance between parents. Although most pollen flow occurred within the population, pollen flow from outside the population accounted for 2.5% of the total. When historical and contemporary pollen flows among populations were compared, the levels of pollen flow seem to have declined recently. We conclude that to conserve M. stellata, it is important to preserve the whole population by maintaining its metapopulation structure and the gene flow among its populations. PMID- 21642216 TI - Sex chromosomes in flowering plants. AB - Sex chromosomes in dioecious and polygamous plants evolved as a mechanism for ensuring outcrossing to increase genetic variation in the offspring. Sex specificity has evolved in 75% of plant families by male sterile or female sterile mutations, but well-defined heteromorphic sex chromosomes are known in only four plant families. A pivotal event in sex chromosome evolution, suppression of recombination at the sex determination locus and its neighboring regions, might be lacking in most dioecious species. However, once recombination is suppressed around the sex determination region, an incipient Y chromosome starts to differentiate by accumulating deleterious mutations, transposable element insertions, chromosomal rearrangements, and selection for male-specific alleles. Some plant species have recently evolved homomorphic sex chromosomes near the inception of this evolutionary process, while a few other species have sufficiently diverged heteromorphic sex chromosomes. Comparative analysis of carefully selected plant species together with some fish species promises new insights into the origins of sex chromosomes and the selective forces driving their evolution. PMID- 21642217 TI - Multiscale analysis of Hymenocallis coronaria (Amaryllidaceae) genetic diversity, genetic structure, and gene movement under the influence of unidirectional stream flow. AB - Understanding gene movement patterns in unidirectional flow environments and their effect on patterns of genetic diversity and genetic structure is necessary to manage these systems. Hypotheses and models to explain genetic patterns in streams are rare, and the results of macrophyte studies are inconsistent. This study addresses Ritland's (Canadian Journal of Botany 67: 2017-2024) unidirectional diversity hypothesis, the one-dimensional stepping stone model, and the metapopulation model within and among populations. Hymenocallis coronaria, an aquatic macrophyte of rocky river shoals of the SE USA, was sampled in four river basins. Within populations and among populations <16.2 km apart had significant isolation by distance. However, the rate of gene flow decay was not consistent with a one-dimensional stepping stone model, nor was evidence strong or consistent for Ritland's hypothesis. Some evidence indicates that localized metapopulation processes may be affecting genetic diversity and structure; however, gene flow patterns inconsistent with the assumptions of the linear and unidirectional models are also a possible influence. We discuss three variants on the one-dimensional stepping stone model. Future research in linear environments should examine the expectations of these models. This study is also one of the first efforts to calculate population genetic parameters using a new program, TETRASAT. PMID- 21642218 TI - Testing implicit assumptions regarding the age vs. size dependence of stem biomechanics using Pittocaulon (Senecio) praecox (Asteraceae). AB - Strong covariation between organismal traits is often taken as an indication of a potentially adaptively significant relationship. Because one of the main functions of woody stems is mechanical support, identifying the factors that covary with biomechanics is essential for inference of adaptation. To date in such studies, stem biomechanics is plotted against stem age or size, thus with implicit assumptions regarding the importance of each in determining mechanics. Likewise, comparing ontogenies between individuals is central to the study of ontogenetic evolution (e.g., heterochrony). Both absolute age and size have been used, but the rationale for choosing one over the other has not been examined. Sampling a plant of simple architecture across microsites with differing sizes for the same absolute age, we compared regressions of stem length, mechanics, and tissue areas against age and size. Stem length was predicted by diameter but not by age, and stem biomechanics and tissue areas were better explained by stem length rather than age. We show that the allometric and mechanical properties observed across microsites are uniform despite great plasticity in other features (e.g., size and wood anatomy) and suggest that this uniformity is an example of developmental homeostasis. Finally, we discuss reasons for preferring size over absolute age as a basis for comparing ontogenies between individuals. PMID- 21642219 TI - Inferring phylogeny at low taxonomic levels: utility of rapidly evolving cpDNA and nuclear ITS loci. AB - Plant molecular systematic studies of closely related taxa have relied heavily on sequence data from nuclear ITS and cpDNA. Positive attributes of using ITS sequence data include the rapid rate of evolution compared to most plastid loci and availability of universal primers for amplification and sequencing. On the other hand, ITS sequence data may not adequately track organismal phylogeny if concerted evolution and high rDNA array copy number do not permit identification of orthologous copies. Shaw et al. (American Journal of Botany 92: 142-166) recently identified nine plastid regions that appear to provide more potentially informative characters than many other plastid loci. In the present study, sequences of these loci and ITS were obtained for six taxonomic groups in which phylogenetic relationships have been difficult to establish using other data. The relative utility of these regions was compared by assessing the number of parsimony informative characters, character congruence, resolution of inferred trees, clade support, and accuracy. No single locus emerged as the best in all lineages for any of these measures of utility. Results further indicated that in preliminary studies, sampling strategy should include at least four exemplar taxa. The importance of sampling data from independent distributions is also discussed. PMID- 21642220 TI - Conservation genetics of an endemic and endangered epiphytic Laelia speciosa (Orchidaceae). AB - We used isozymes (16 loci in 11 enzymatic systems) from Laelia speciosa, an endemic and endangered epiphytic orchid of Mexico, to assess the genetic diversity and population genetic structure in nine populations distributed along its geographic range, as well as to detect those populations that are genetically unique and therefore deserve high-priority protection. On average, the genetic diversity was high (percentage of polymorphic loci, P(p) = 76%, mean number of alleles per locus, A = 3.34, the average observed heterozygosity H(O) = 0.302, the average expected heterozygosity H(E) = 0.382). Moderate levels of inbreeding ( f = 0.216, 95% confidence interval = 0.029-0.381) were found. Low levels of genetic differentiation were observed among populations ((p) = 0.040); however, there was a significant correlation between geographic and genetic distances among the populations (Mantel test: r(2) = 0.43, P < 0.05). Populations located within the same mountain range were genetically more similar. Private alleles were found, so proper management requires protection and maintenance of genetic diversity throughout its range. In case of reintroduction, we suggest using individuals propagated from seeds from as many capsules as possible, from close populations. An ex situ conservation strategy also is proposed. PMID- 21642221 TI - Uniform genetic diversity, low differentiation, and neutral evolution characterize contemporary refuge populations of Taiwan fir (Abies kawakamii, Pinaceae). AB - Based on fossil pollen, the distribution range of Taiwan fir [Abies kawakamii (Hay.) Ito] (Pinaceae) is smaller than it was 50 000 years ago. To characterize the present refuge populations of A. kawakamii, which survive only in subalpine forests in Taiwan, we surveyed nuclear genes and chloroplast intergenic spacers to assess the genetic diversity of Taiwan fir. Populations maintain high genetic diversity and contain similar numbers of haplotypes for the GapC (cytosolic glyceraldehyde 3-phosphate dehydrogenase) fragment. Haplotypes for GapC are generally widespread, and population-specific haplotypes accounted for 2.5% of the total. Differentiation among populations is very low (G(ST) = 0.01). Only three haplotypes were detected for the cpDNA marker, and every population had one or two haplotypes. In a neutrality test, the variation in nucleotides did not deviate from that expected with neutral evolution for either marker. A retreat route to higher elevations was not evident from either the GapC or cpDNA markers. Hsuehshan was the site of the most divergent population in Taiwan. We concluded that uniform genetic diversity, low differentiation, low numbers of population specific haplotypes, and neutral evolution characterize contemporary refuge populations of Taiwan fir. PMID- 21642222 TI - Visitation, effectiveness, and efficiency of 15 genera of visitors to wild radish, Raphanus raphanistrum (Brassicaceae). AB - Plant-pollinator interactions are one of the most important and variable mutualisms in nature. Multiple pollinators often visit plants and can vary in visitation rates, pollen removal and deposition, and spatial and temporal distribution, altering plant reproduction and patterns of pollinator-mediated selection. Although some visitors may not be effective pollinators, pollinator effectiveness is rarely estimated directly as seed set resulting from a single visit by each taxon visiting generalist plants. For two years, effectiveness of visitors to wild radish, Raphanus raphanistrum, was quantified by counting seeds set and pollen grains removed as a result of a single visit. We calculated a pollinator's importance to plant reproduction as the product of visitation rate and single-visit seed set, and regressed pollinator body size on pollen-removal and on seed set effectiveness. Although pollinators differed in effectiveness and visitation rates, pollinator importance was primarily determined by visitation rates. In contrast to similar 2-yr studies, pollinator assemblage composition varied little, suggesting pollinator-mediated selection can be consistent across years for this generalist. Larger pollinators were more effective than smaller at effecting seed set, but body size was a poor predictor of pollen removal ability. Instead, pollen-removal effectiveness may be more influenced by foraging behavior than size. PMID- 21642223 TI - The origin and number of introductions of the Hawaiian endemic Silene species (Caryophyllaceae). AB - The Hawaiian endemic Silene are a small group of woody or semiwoody representatives from a large, predominantly herbaceous, species-rich genus. We here investigated the origin and number of introductions of the endemic Hawaiian Silene based on phylogenetic relationships inferred from DNA sequences from both the plastid (the rps16 intron) and the nuclear (ribosomal internal transcribed sequences, ITS, and intron 23 of the RPB2 gene) genomes. Silene antirrhina, a widespread weedy American annual, is strongly supported as sister to a monophyletic group consisting of the Hawaiian Silene, indicating a single colonization event. There are no obvious morphological similarities between S. antirrhina and any of the species of Hawaiian Silene. Our results suggest an American origin for the Hawaiian endemics because that would require only a single trans-ocean dispersal. Two of the Hawaiian endemics (S. struthioloides and S. hawaiiensis) that form a subclade in the analyses have evolved woodiness after introduction to the Hawaiian Islands. Our results contribute to other recent results based on molecular phylogenetics that emphasize the American continent as a source area for the Hawaiian flora and support a striking morphological radiation and evolution of woodiness from a single introduction to the archipelago. PMID- 21642224 TI - Environmental effects on pollen-pistil compatibility between Phlox cuspidata and P. drummondii (Polemoniaceae): implications for hybridization dynamics. AB - Postpollination mechanisms of reproductive isolation can critically influence the amount of gene flow between hybridizing species. While much evidence exists for genetically based pollen-pistil incompatibility, we show that environmental variation also influences the postpollination performance of heterospecific pollen in the annual Phlox hybrid system. Thus, the environmental segregation of species can influence hybridization dynamics. We found that P. cuspidata was restricted to soils of low Ca concentrations in the field and performed better under experimentally low Ca; P. drummondii was able to inhabit high-Ca soils and sometimes performed better in this environment. To determine whether soil Ca influenced pollen-pistil compatibility in a manner that alters pollen siring success, single-donor pollinations were performed in a completely factorial crossing design between species, maternal Ca environments, and paternal Ca environments. Maternal and paternal environments interacted in their effects on pollen-pistil compatibility for both inter- and intraspecific crosses, such that pollen performance was highest when mothers and fathers were grown in different soil Ca environments. These results suggest that when Phlox species predictably inhabit different environments, environmental heterogeneity can impede the processes of speciation and local adaptation by enhancing the performance of pollen dispersed across species and environments. PMID- 21642225 TI - Reproductive ecology of a federally endangered legume, Baptisia arachnifera, and its more widespread congener, B. lanceolata (Fabaceae). AB - Comparisons between rare species and their more common congener species can provide valuable information for conservation. Reproductive traits have previously been shown to be critical for reproductive success and persistence of rare species. In this study, we compared floral, seedpod, and seed traits of two Baptisia species (one endangered and one common) to assess differences in reproduction between species and among populations. Because heat can trigger germination in hard-seeded legumes, we also exposed Baptisia seeds to a range of high temperatures (60-100 degrees C) and determined seed viability. The rare Baptisia arachnifera had significantly greater pod damage by insects and produced significantly fewer, yet heavier, seeds than B. lanceolata. While B. arachnifera seeds were seldom viable at temperatures above 80 degrees C, approximately 40% of B. lanceolata seeds maintained viability up to 100 degrees C. Our various seed trait measurements suggest that B. arachnifera may be a poorer colonizer than the more widespread B. lanceolata. Additionally, B. arachnifera's reduced tolerance for high temperatures may have implications for appropriate fire management regimes for this endangered species. PMID- 21642226 TI - Adaptive radiation of shrubby tarweeds (Deinandra) in the California Islands parallels diversification of the Hawaiian silversword alliance (Compositae Madiinae). AB - Phylogenetic analyses of nuclear rDNA transcribed spacers and cytogenetic studies of interspecific hybrids reported here uphold Carlquist's hypothesis (1965, Island Biology) that shrubby tarweeds (Deinandra) of Guadalupe Island, Mexico, are products of in situ radiation in the California Islands, where evidence of plant diversification has been equivocal. Based on the rDNA findings, the Guadalupe Island endemics (D. frutescens, D. greeneana subsp. greeneana, and D. palmeri) constitute a clade that arose since the late Pliocene, well after the origin of Guadalupe Island and diversification of annual, mainland Californian lineages of Deinandra. High interfertility and normal meiosis in F(1) hybrids between the three endemics contrast with reduced interfertility (to complete intersterility) and meiotic irregularities in F(1) hybrids between other, mostly mainland species of Deinandra. Cloned rDNA sequences provided no convincing evidence of introgression among the Guadalupe Island deinandras; morphological, phenological, and/or habitat differences among those taxa indicate ecological barriers to gene flow and a probable role for ecological divergence in diversification. Biosystematic and molecular phylogenetic data for shrubby tarweeds of Guadalupe Island and another secondarily woody, oceanic-island tarweed lineage, the Hawaiian silversword alliance, reveal strikingly similar evolutionary histories. Both groups violate Baker's Rule by stemming from self incompatible ancestors in western North America, and each has undergone within island diversification without evolution of strong sterility barriers among lineages. Evolutionary parallels between these Hawaiian and California Island lineages of Madiinae, first suggested by Carlquist, may reflect characteristics of tarweeds that facilitate insular colonization and adaptive radiation. PMID- 21642227 TI - Colored floral organs influence pollinator behavior and pollen transfer in Commelina communis (Commelinaceae). AB - Visual floral guides such as colored anthers, lines, dots, and UV-absorption patterns on petals are commonly observed in insect-pollinated angiosperms. Floral guides that are known to enhance foraging efficiency of visitors on flowers thus promote return visits (foraging facilitation hypothesis, which predicts that visitors will discriminate against flowers with inferior floral guides). In this study, we experimentally examined the hypothesis that floral guides also prevent pollen-theft behavior by floral visitors (theft prevention hypothesis), which has rarely been tested. Nectarless flowers of Commelina communis have three types of brightly colored floral organs: large blue petals, rewarding yellow anthers, and nonrewarding yellow anthers. Colored floral organs were removed artificially from plants in two natural populations of C. communis. Removal of the nonrewarding yellow anthers diminished hoverings in front of flowers and tended to reduce the number of total floral visitor landings, supporting the foraging facilitation hypothesis. Additionally, removal of the rewarding yellow anther decreased the frequency of legitimate landings on flowers and the legitimate landing-to-total landing ratio, which is consistent with the theft prevention hypothesis. The nonrewarding anthers and the rewarding yellow anthers were shown to play an important role in increasing visitor landings and orienting floral visitors toward a landing point appropriate for pollination, respectively. We also showed that the absence of yellow anthers decreased both pollen dispatch from brown anthers and receipt by stigmas in C. communis. These findings support both the foraging facilitation hypothesis and the theft prevention hypothesis. PMID- 21642228 TI - Duodichogamy and androdioecy in the Chinese Phyllanthaceae Bridelia tomentosa. AB - Flowering plants commonly separate male and female function in time, but rarely are the two stages synchronized within and among individuals. One such temporal mating system is duodichogamy in which each plant produces two batches of male flowers that are temporally separated by a batch of female flowers, with within individual synchrony and among-individual asynchrony to ensure mating partners. Duodichogamy is known only from a few species in four genera in unrelated families. We report on duodichogamy in the Chinese tree species Bridelia tomentosa (Phyllanthaceae), a common colonizer of disturbed habitats. In three populations monitored over 2 yr, most trees flowered in the order male -> female > male, and resting periods between flowering bouts precluded selfing almost completely. Individuals flowered for several weeks, with the onset of flowering slightly asynchronous among trees. Pollination was by flies, and experimental pollen supplementation of a subset of a tree's flowers did not increase fruit set, suggesting high levels of insect visitation and possible resource limitation. Nineteen percent of the 166 trees monitored skipped the first male phase, and another 13% skipped the female phase, remaining male in both years (and also a third year). The regular presence of pure males, if genetically fixed, would make B. tomentosa androdioecious in addition to duodichogamous. Comparison of duodichogamous taxa known so far shows that all have few ovules, fitting with the hypothesis that duodichogamy may result from male competition for access to a small supply of ovules. PMID- 21642229 TI - How robust is nonrandom mating in wild radish: do small pollen loads coupled with more competing pollen donors lead to random mating? AB - In previous studies of the weedy annual Raphanus sativus we have demonstrated that mating is nonrandom in greenhouse plants, suggesting that sexual selection is possible. To investigate how these greenhouse results might translate to conditions more similar to the field, we manipulated both pollen load size and the number of competing pollen donors on stigmas. While the smallest pollen loads (22 grains per stigma) were small enough to reduce fruit and seed set, seed siring success was unaffected by pollen load size. When the number of competing donors in a mixed pollination was increased to four, the proportion of seeds sired by the pollen donors was the furthest from expectation, suggesting that nonrandom mating increases as the number of donors per pollination increases. There was no significant interaction between pollen load size and number of competitors per pollination. Overall, mating remained nonrandom across all treatments. Thus differential seed paternity is likely to occur in the field as well as in the greenhouse. PMID- 21642231 TI - Successful treatment of infantile subglottic hemangioma with oral propranolol. PMID- 21642230 TI - Paracrine and endocrine effects of adipose tissue on cancer development and progression. AB - The past few years have provided substantial evidence for the vital role of the local tumor microenvironment for various aspects of tumor progression. With obesity and its pathophysiological sequelae still on the rise, the adipocyte is increasingly moving center stage in the context of tumor stroma-related studies. To date, we have limited insight into how the systemic metabolic changes associated with obesity and the concomitant modification of the paracrine and endocrine panel of stromal adipocyte-derived secretory products ("adipokines") influence the incidence and progression of obesity-related cancers. Here, we discuss the role of adipocyte dysfunction associated with obesity and its potential impact on cancer biology. PMID- 21642232 TI - Preoperative assessment for children requiring dental treatment under general anesthesia. AB - OBJECTIVE: This study aimed to describe children <6 years requiring general anesthesia for dental treatment and factors associated with a change in medical management prior to surgery. STUDY DESIGN: This case series reviewed the past medical history and preoperative assessment of patients referred for dental preoperative evaluations at a single institution (2005-2008). A "deflection" was defined as a recommendation to change preoperative or operative care based on the preoperative assessment. The sample was analyzed using descriptive, bivariate, and multivariate analyses. RESULTS: Of 648 subjects (aged 9 months to 6 years, mean 3.9 years), 63% had a past medical history abnormality and 38% had previous surgery. In total, 14% were deflected, most commonly because of the addition of infective endocarditis prophylaxis (29%). A history of coagulation disorder had the strongest association with deflection (P < .0001, odds ratio = 10.0, 95% confidence interval = 4.6-22.1), followed by cardiac anomalies. CONCLUSION: Preoperative assessments for pediatric dental treatment frequently identify medical problems resulting in treatment plan alterations. PMID- 21642233 TI - Simplification of a light-based model for estimating final internode length in greenhouse cucumber canopies. AB - BACKGROUND AND AIMS: Light quantity and quality affect internode lengths in cucumber (Cucumis sativus), whereby leaf area and the optical properties of the leaves mainly control light quality within a cucumber plant community. This modelling study aimed at providing a simple, non-destructive method to predict final internode lengths (FILs) using light quantity and leaf area data. METHODS: Several simplifications of a light quantity and quality sensitive model for estimating FILs in cucumber have been tested. The direct simplifications substitute the term for the red : far-red (R : FR) ratios, by a term for (a) the leaf area index (LAI, m(2) m(-2)) or (b) partial LAI, the cumulative leaf area per m(2) ground, where leaf area per m(2) ground is accumulated from the top of each plant until a number, n, of leaves per plant is reached. The indirect simplifications estimate the input R : FR ratio based on partial leaf area and plant density. KEY RESULTS: In all models, simulated FILs were in line with the measured FILs over various canopy architectures and light conditions, but the prediction quality varied. The indirect simplification based on leaf area of ten leaves revealed the best fit with measured data. Its prediction quality was even higher than of the original model. CONCLUSIONS: This study showed that for vertically trained cucumber plants, leaf area data can substitute local light quality data for estimating FIL data. In unstressed canopies, leaf area over the upper ten ranks seems to represent the feedback of the growing architecture on internode elongation with respect to light quality. This highlights the role of this domain of leaves as the primary source for the specific R : FR signal controlling the final length of an internode and could therefore guide future research on up-scaling local processes to the crop level. PMID- 21642234 TI - Heterogeneity of pathological processes in amyotrophic lateral sclerosis? PMID- 21642235 TI - Increased glutamine in leaves of poplar transgenic with pine GS1a caused greater anthranilate synthetase alpha-subunit (ASA1) transcript and protein abundances: an auxin-related mechanism for enhanced growth in GS transgenics? AB - The initial reaction in the pathway leading to the production of indole-3-acetic acid (IAA) in plants is the reaction between chorismate and glutamine to produce anthranilate, catalysed by the enzyme anthranilate synthase (ASA; EC 4.1.3.27). Compared with non-transgenic controls, leaves of transgenic poplar with ectopic expression of the pine cytosolic glutamine synthetase (GS1a; EC 6.3.1.2) produced significantly greater glutamine and significantly enhanced ASA alpha-subunit (ASA1) transcript and protein (approximately 130% and 120% higher than in the untransformed controls, respectively). Similarly, tobacco leaves fed with 30 mM glutamine and 2 mM chorismate showed enhanced ASA1 transcript and protein (175% and 90% higher than controls, respectively). Furthermore, free IAA was significantly elevated both in leaves of GS1a transgenic poplar and in tobacco leaves fed with 30 mM glutamine and 2 mM chorismate. These results indicated that enhanced cellular glutamine may account for the enhanced growth in GS transgenic poplars through the regulation of auxin biosynthesis. PMID- 21642236 TI - Roles of xanthophylls and exogenous ABA in protection against NaCl-induced photodamage in rice (Oryza sativa L) and cabbage (Brassica campestris). AB - Changes in actual efficiency of PS II photochemistry, non-photochemical quenching (NPQ), content of xanthophylls and kinetics of de-epoxidation were studied in ABA fed and non-ABA-fed leaves of rice and cabbage under NaCl stress. Salt stress induced more progressive decrease in actual efficiency of PS II photochemistry (FPS II), higher reduction state of PS II, and a small significant increase in NPQ in NaCl-sensitive rice plants as compared with NaCl-tolerant cabbage plants, whereas exogenously supplied ABA alleviated the decrease in actual efficiency of PS II photochemistry (FPS II), induced a lower reduction state of PS II, and caused higher capacity of NPQ in ABA-fed plants than in non-ABA-fed plants. As a result, there were higher activities of photosynthetic electron transport, higher capacity of energy dissipation, and lower cumulation of excess light in cabbage than in rice plants, and in ABA-fed leaves than in non-ABA-fed leaves. The effect of ABA was more efficient in cabbage than in rice plants. Addition of exogenous ABA resulted in enhancement of the size of the xanthophyll cycle pool, promotion of de-epoxidation of the xanthophyll cycle components, and a rise in the level of NPQ by altering the kinetics of de-epoxidation of the xanthophyll cycle. Protection from photodamage appears to be achieved by coordinated contributions by exogenous ABA and xanthophyll cycle-mediated NPQ. This variety of photoprotective mechanisms may be essential for conferring photodamage tolerance under NaCl stress. PMID- 21642237 TI - Equilibrative nucleoside transporter 1 (ENT1) is critical for pollen germination and vegetative growth in Arabidopsis. AB - ENT1 of Arabidopsis thaliana was the first member of the equilibrative nucleoside transporter (ENT) family to be identified in plants and characterized as a cellular, high-affinity nucleoside importer. Evidence is presented here for a tonoplast localization of ENT1 based on proteome data and Western blot analyses. Increased export of adenosine from reconstituted tonoplast preparations from 35S:ENT1 mutants compared with those from the wild type and ENT1-RNAi mutants support this view. Furthermore, increased vacuolar adenosine and vacuolar 2'3' cAMP (an intermediate of RNA catabolism) contents in ENT1-RNAi mutants, but decreased contents of these metabolites in 35S:ENT1 over-expresser mutants, were observed. An up-regulation of the salvage pathway was detected in the latter mutants, leading to the conclusion that draining the vacuolar adenosine storage by ENT1 over-expression interferes with cellular nucleotide metabolism. As a consequence of the observed metabolic alterations 35S:ENT1 over-expresser mutants exhibited a smaller phenotypic appearance compared with wild-type plants. In addition, ENT1:RNAi mutants exhibited significantly lower in vitro germination of pollen and contained reduced internal and external ATP levels. This indicates that ENT1-mediated nucleosides, especially adenosine transport, is important for nucleotide metabolism, thus influencing growth and pollen germination. PMID- 21642238 TI - Predictors of low-care prevalence in Florida nursing homes: the role of Medicaid waiver programs. AB - PURPOSE OF THE STUDY: To examine the relationship between county-level Medicaid home- and community-based service (HCBS) waiver expenditures and the prevalence of low-care residents in Florida nursing homes (NHs). DESIGN AND METHODS: The present study used a cross-sectional design. We combined two data sources: NH facility-level data (including characteristics of the facility and its residents) and county-level market characteristics (including HCBS waiver expenditures) for 653 Florida NHs in 2007. Low-care was defined as residents who require no physical assistance in any of the 4 late-loss activities of daily living (bed mobility, toileting, transferring, and eating). We estimated a 2-level hierarchical linear model (HLM) to examine the relationship between Medicaid HCBS waiver expenditures and the prevalence of low-care residents while accounting for resident assessment, facility-, and county-level covariates. RESULTS: All Florida counties offered 2 statewide waivers, and 33 counties offered one or more of the 4 regional Medicaid HCBS waivers in 2007. Per-month beneficiary expenditures ranged from $755 to $1,778. The average Florida NH had 120 beds, and 8.0% of its residents were classified as low-care. Results from the HLM model showed that a $10,000 increase in per-enrollee HCBS waiver expenditures was associated with a 3.5 percentage point reduction in low-care resident prevalence (p = .03). IMPLICATIONS: The findings suggest that Medicaid HCBS waiver programs may reduce the prevalence of low-care residents in NHs. Future studies should evaluate whether Medicaid HCBS waiver programs are effective in promoting community-living among low-care residents and mitigating the growth in long-term care expenditures. PMID- 21642239 TI - Plasticity of hatching in amphibians: evolution, trade-offs, cues and mechanisms. AB - Many species of frogs and salamanders, in at least 12 families, alter their timing of hatching in response to conditions affecting mortality of eggs or larvae. Some terrestrially laid or stranded embryos wait to hatch until they are submerged in water. Some embryos laid above water accelerate hatching if the eggs are dehydrating; others hatch early if flooded. Embryos can hatch early in response to predators and pathogens of eggs or delay hatching in response to predators of larvae; some species do both. The phylogenetic pattern of environmentally cued hatching suggests that similar responses have evolved convergently in multiple amphibian lineages. The use of similar cues, including hypoxia and physical disturbance, in multiple contexts suggests potential shared mechanisms underlying the capacity of embryos to respond to environmental conditions. Shifts in the timing of hatching often have clear benefits, but we know less about the trade-offs that favor plasticity, the mechanisms that enable it, and its evolutionary history. Some potentially important types of cued hatching, such as those involving embryo-parent interactions, are relatively unexplored. I discuss promising directions for research and the opportunities that the hatching of amphibians offers for integrative studies of the mechanisms, ecology and evolution of a critical transition between life-history stages. PMID- 21642240 TI - Molecular genetic and functional characterization implicate muscle-restricted coiled-coil gene (MURC) as a causal gene for familial dilated cardiomyopathy. AB - BACKGROUND: Dilated cardiomyopathy (DCM) and hypertrophic cardiomyopathy (HCM) are classic forms of systolic and diastolic heart failure, respectively. Mutations in genes encoding sarcomere and cytoskeletal proteins are major causes of HCM and DCM. MURC, encoding muscle-restricted coiled-coil, a Z-line protein, regulates cardiac function in mice. We investigated potential causal role of MURC in human cardiomyopathies. METHODS AND RESULTS: We sequenced MURC in 1199 individuals, including 383 probands with DCM, 307 with HCM, and 509 healthy control subjects. We found 6 heterozygous DCM-specific missense variants (p.N128K, p.R140W, p.L153P, p.S307T, p.P324L, and p.S364L) in 8 unrelated probands. Variants p.N128K and p.S307T segregated with inheritance of DCM in small families (chi(2)=8.5, P=0.003). Variants p.N128K, p.R140W, p.L153P, and p.S364L were considered probably or possibly damaging. Variant p.P324L recurred in 3 independent probands, including 1 proband with a TPM1 mutation (p.M245T). A deletion variant (p.L232-R238del) was present in 3 unrelated HCM probands, but it did not segregate with HCM in a family who also had a MYH7 mutation (p.L907V). The phenotype in mutation carriers was notable for progressive heart failure leading to heart transplantation in 4 patients, conduction defects, and atrial arrhythmias. Expression of mutant MURC proteins in neonatal rat cardiac myocytes transduced with recombinant adenoviruses was associated with reduced RhoA activity, lower mRNA levels of hypertrophic markers and smaller myocyte size as compared with wild-type MURC. CONCLUSIONS: MURC mutations impart loss-of-function effects on MURC functions and probably are causal variants in human DCM. The causal role of a deletion mutation in HCM is uncertain. PMID- 21642241 TI - Increased microRNA-1 and microRNA-133a levels in serum of patients with cardiovascular disease indicate myocardial damage. AB - BACKGROUND: Recently, elevation of circulating muscle-specific microRNA (miRNA) levels has been reported in patients with acute myocardial infarction. However, it is still unclear from which part of the myocardium or under what conditions miRNAs are released into circulating blood. The purpose of this study was to identify the source of elevated levels of circulating miRNAs and their function in cardiovascular diseases. METHODS AND RESULTS: Serum levels of miRNA (miR)-1 and miR-133a were increased significantly in patients not only with acute myocardial infarction but also with unstable angina pectoris and Takotsubo cardiomyopathy without elevation of serum creatine phosphokinase or cardiac troponin. MicroRNA microarray analysis of the heart from a mouse model of myocardial infarction indicated that the levels of miR-1, miR-133a, miR-208a, and miR-499 were significantly reduced in the infarcted myocardium. In situ hybridization of miR-133a also showed that miR-133a levels were very low in the infarcted and peri-infarcted myocardium. It has been shown that circulating miRNAs are localized inside exosomes, which are released after Ca(2+) stimulation. We stimulated H9c2 cardiomyoblasts with A23187 and measured miR-133a levels in the exosome fraction of the culture medium. A23187 induced a dose dependent release of miR-133a, and significant elevation was observed only at concentrations where dead cells were detected. We also found that miR-133a containing exosomes reduced the luciferase activity of 293FT cells transfected with an miR-133a sensor vector. CONCLUSIONS: These results suggest that elevated levels of circulating miR-133a in patients with cardiovascular diseases originate mainly from the injured myocardium. Circulating miR-133a can be used as a marker for cardiomyocyte death, and it may have functions in cardiovascular diseases. PMID- 21642242 TI - Upper arm intermittent ischaemia reduces exercise-related increase of platelet reactivity in patients with obstructive coronary artery disease. AB - OBJECTIVE: To assess whether upper arm ischaemia influences exercise-induced myocardial ischaemia and platelet activation in patients with coronary artery disease (CAD). DESIGN: Crossover study. SETTING: University hospital. PATIENTS: Twenty patients (17 men) of mean+/-SD age 64+/-8 years with stable CAD. INTERVENTIONS: Patients underwent two exercise stress tests (ESTs) on two separate days in a randomised manner: (1) a maximal EST only (EST-1); (2) a maximal EST after intermittent upper arm ischaemia (cycles of alternating 5-min inflation and 5-min deflation of a standard blood pressure cuff) (EST-2). Blood samples were obtained to evaluate platelet reactivity. MAIN OUTCOME MEASURES: Platelet reactivity was assessed by flow cytometry at rest and after EST, with and without ADP stimulation, by measuring the percentage of monocyte-platelet aggregates (MPAs) and CD41 platelet expression measured as mean fluorescence intensity. RESULTS: Remote ischaemia had no significant effect on EST-induced myocardial ischaemia. At rest there were no differences before EST-1 and EST-2 in basal MPA (20.7+/-2.3 vs 20.8+/-2.4, p=0.56) and CD41 (21.5+/-2.3 vs 21.3+/-2.3, p=0.39), and ADP stimulation induced a similar increase in both MPA (+15.2+/-8.2% vs +14.9+/-8.4%, p=0.71) and CD41 (+15.7+/-5.7% vs 13.37+/-6.9%, p=0.59). While no differences in the increase in MPA and CD41 expression were observed after EST 1 and EST-2, ADP stimulation after EST-2 induced a lower increase in MPA (+18.3+/ 8.1% vs +27.9+/-9.7%, p<0.001) and CD41 (+18.3+/-9.2% vs +27.2+/-12.4%, p<0.001) than after EST-1. CONCLUSION: These results show that, in patients with stable CAD, remote ischaemia induces protection against an exercise-related increase in platelet reactivity. PMID- 21642243 TI - A complication of the use of an intra-osseous needle. PMID- 21642244 TI - Gender and mortality following hospitalisation for COPD. PMID- 21642245 TI - A trial of intrapleural adenoviral-mediated Interferon-alpha2b gene transfer for malignant pleural mesothelioma. AB - New therapeutic strategies are needed for malignant pleural mesothelioma (MPM). We conducted a single-center, open-label, nonrandomized, pilot and feasibility trial using two intrapleural doses of an adenoviral vector encoding human IFN alpha (Ad.IFN-alpha2b). Nine subjects were enrolled at two dose levels. The first three subjects had very high pleural and systemic IFN-alpha concentrations resulting in severe "flu-like" symptoms necessitating dose de-escalation. The next six patients had reduced (but still significant) pleural and serum IFN-alpha levels, but with tolerable symptoms. Repeated vector administration appeared to prolong IFN-alpha expression levels. Anti-tumor humoral immune responses against mesothelioma cell lines were seen in seven of the eight subjects evaluated. No clinical responses were seen in the four subjects with advanced disease. However, evidence of disease stability or tumor regression was seen in the remaining five patients, including one dramatic example of partial tumor regression at sites not in contiguity with vector infusion. These data show that Ad.IFN-alpha2b has potential therapeutic benefit in MPM and that it generates anti-tumor immune responses that may induce anatomic and/or metabolic reductions in distant tumor. Clinical trial registered with www.clinicaltrials.gov (NCT 01212367). PMID- 21642246 TI - Intersections between pulmonary development and disease. AB - Recent advances in cellular, molecular, and developmental biology have revolutionized our concepts regarding the process of organogenesis that have important implications for our understanding of both lung formation and pulmonary disease pathogenesis. Pulmonary investigators have long debated whether developmental processes are recapitulated during normal repair of the lung or in the setting of chronic pulmonary diseases. Although the cellular events involved in lung morphogenesis and those causing pulmonary disease are likely to include processes that are distinct, there is increasing evidence that the pathogenesis of many lung disorders involves the same genetic machinery that regulates cell growth,specification, and differentiation during normal lung development. PMID- 21642247 TI - First- and second-trimester fetal size and asthma outcomes at age 10 years. AB - RATIONALE: Greater early fetal size is associated with reduced asthma risk and improved lung function in early childhood. OBJECTIVES: To test the hypothesis that associations between early fetal size, asthma symptoms, and lung function persist into later childhood. METHODS: In a longitudinal study, first- and second trimester fetal measurements were recorded. At 10 years of age a respiratory questionnaire was completed. Spirometry, bronchial challenge, and skin-prick testing were undertaken in a subset. MEASUREMENTS AND MAIN RESULTS: Fetal measurements were available in the first trimester for 853 individuals and the second trimester for 1,453. Questionnaires were returned for 927 children and 449 underwent detailed phenotyping. For each millimeter increase in first trimester size, asthma risk reduced by 6% (95% confidence interval[CI], 1-11) and FEV1 was higher by an average of 6 ml (95% CI, 1-11).First-trimester size was reduced in those with asthma at both 5 and 10 years compared with early or late onset wheeze (P , 0.02). Compared with persistent high growth in first and second trimesters,persistent low growth was associated with increased asthma risk(odds ratio, 2.8; 95% CI, 1.2-6.9) and a mean reduction in FEV1 of 103 ml (95% CI, 13 194), whereas increasing fetal size was associated with increased eczema risk (odds ratio, 2.5; 95% CI, 1.2-5.3). CONCLUSIONS: Reduced fetal size from the first trimester is associated with increased risk for asthma and obstructed lung function in childhood. Relative change in size after the first trimester is associated with eczema. PMID- 21642248 TI - Case-control study of lung function in World Trade Center Health Registry area residents and workers. AB - RATIONALE: Residents and area workers who inhaled dust and fumes from the World Trade Center disaster reported lower respiratory symptoms in two World Trade Center Health Registry surveys (2003-2004 and 2006-2007), but lung function data were lacking. OBJECTIVES: To examine the relationship between persistent respiratory symptoms and pulmonary function in a nested case-control study of exposed adult residents and area workers 7-8 years after September 11, 2001. METHODS: Registrants reporting post September 11th onset of a lower respiratory symptom in the first survey and the same symptom in the second survey were solicited as potential cases. Registrants without lower respiratory symptoms in either Registry survey were solicited as potential control subjects. Final case control status was determined by lower respiratory symptoms at a third interview (the study), when spirometry and impulse oscillometry were also performed. MEASUREMENTS AND MAIN RESULTS: We identified 180 cases and 473 control subjects. Cases were more likely than control subjects to have abnormal spirometry (19% vs. 11%; P < 0.05), and impulse oscillometry measurements of elevated airway resistance (R5; 68% vs. 27%; P < 0.0001) and frequency dependence of resistance (R5-20; 36% vs. 7%; P < 0.0001). When spirometry was normal, cases were more likely than control subjects to have elevated R5 and R5-20 (62% vs. 25% and 27% vs. 6%, respectively; both P < 0.0001). Associations between symptoms and oscillometry held when factors significant in bivariate comparisons (body mass index, spirometry, and exposures) were analyzed using logistic regression. CONCLUSIONS: This study links persistent respiratory symptoms and oscillometric abnormalities in World Trade Center-exposed residents and area workers. Elevated R5 and R5-20 in cases despite normal spirometry suggested distal airway dysfunction as a mechanism for symptoms. PMID- 21642249 TI - CD34 is required for dendritic cell trafficking and pathology in murine hypersensitivity pneumonitis. AB - RATIONALE: Although recent work has shown that CD34 plays an important role in the trafficking of inflammatory cells during Th2-biased inflammatory responses, its role in Th1/Th17-biased disease as well as dendritic cell (DC) trafficking is unknown. OBJECTIVES: We used CD34-deficient mice (Cd34(-/-)) to investigate the role of CD34 in the Th1/Th17-biased lung inflammatory disease, hypersensitivity pneumonitis (HP). METHODS: HP was induced in wild-type (wt) and Cd34(-/-) mice by repeated intranasal administration of Saccharopolyspora rectivirgula antigen. Lung inflammation was assessed by histology and analysis of bronchoalveolar lavage cells. Primary and secondary immune responses were evaluated by cytokine recall responses of pulmonary inflammatory cells as well as draining lymph node cells. MEASUREMENTS AND MAIN RESULTS: Cd34(-/-) mice were highly resistant to the development of HP and exhibited an inflammatory pattern more reflective of a primary response to S. rectivirgula rather than the chronic lymphocytosis that is typical of this disease. Cytokine recall responses from Cd34(-/-) lymph node cells were dampened and consistent with a failure of antigen-loaded Cd34(-/-) DCs to deliver antigen and prime T cells in the draining lymph nodes. In agreement with this interpretation, adoptive transfer of wt DCs into Cd34(-/-) mice was sufficient to restore normal sensitivity to HP. CD34 was found to be expressed by wt DCs, and Cd34(-/-) DCs exhibited an impaired ability to chemotax toward a subset of chemokines in vitro. Finally, expression of human CD34 in Cd34(-/-) mice restored normal susceptibility to HP. CONCLUSIONS: We conclude that CD34 is expressed by mucosal DCs and plays an important role in their trafficking through the lung and to the lymph nodes. Our data also suggest that CD34 may play a selective role in the efficient migration of these cells to a subset of chemokines. PMID- 21642250 TI - TB/HIV: an orphan disease? PMID- 21642251 TI - Leptin in fibroproliferative acute respiratory distress syndrome: not just a satiety factor. PMID- 21642252 TI - Could you please pass the salt? PMID- 21642253 TI - Who dies from sarcoidosis and why? PMID- 21642254 TI - Just in the "Bik" of time. PMID- 21642255 TI - How tifacogin could not captivate severe community-acquired pneumonia. PMID- 21642256 TI - Update in sleep medicine 2010. PMID- 21642257 TI - Update in pediatric lung disease 2010. PMID- 21642258 TI - Pulmonary emboli detected by endobronchial ultrasound. PMID- 21642259 TI - Acetaminophen and asthma: spurious association? PMID- 21642260 TI - Possible relationship between asbestos exposure and bronchial asthma: a need for clarification. PMID- 21642261 TI - Early-stage lung cancer mimicking pulmonary arteriovenous malformation. PMID- 21642262 TI - Whole-body magnetic resonance imaging for staging metastatic thymic carcinoma. PMID- 21642263 TI - Does hormone replacement therapy cause breast cancer? An application of causal principles to three studies: part 3. The Women's Health Initiative: unopposed estrogen. AB - BACKGROUND: Studies from the Women's Health Initiative have reported an increased risk of breast cancer in users of estrogen plus progestogen. Among users of estrogen alone an increased risk was not observed. OBJECTIVE: To evaluate the evidence for unopposed estrogen. METHODS: In a related article (Part 2) the authors apply generally accepted causal criteria to the findings for estrogen plus progestogen. Here (Part 3) the authors apply the criteria to the findings for unopposed estrogen, as reported in a clinical trial, and in combined data from the trial and an observational study. RESULTS: In the clinical trial, after 7.1 years of follow-up the relative risk (RR) of invasive breast cancer for women assigned to estrogen was 0.77 in an 'intention-to-treat' analysis (95% CI 0.59 1.01) and 0.67 (95% CI 0.47-0.97) in an 'as treated' analysis; after 10.7 years the risk reduction persisted. Time order was correctly specified; detection bias was minimal; in the 'as treated' analysis confounding was unlikely; duration response and internal consistency could be evaluated only to a limited extent because of scanty data; the findings were discordant with increased risks observed in the Collaborative Reanalysis and the Million Women Study; biological plausibility could not be assessed. In the combined analysis, among women who had previously used estrogen soon after the menopause there was no clear evidence of either a reduction or an increase in the risk of breast cancer among women assigned to estrogen during the trial, or among women who were using estrogen in the observational study when follow-up commenced. The combined analysis did not satisfy the criteria of time order, bias, confounding, statistical stability and strength of association, duration-response, and internal consistency; biological plausibility could not be assessed. CONCLUSIONS: The evidence from the clinical trial suggests that unopposed estrogen does not increase the risk of breast cancer, and may even reduce it. The latter possibility, however, is based on statistically borderline evidence. PMID- 21642264 TI - Does hormone replacement therapy cause breast cancer? An application of causal principles to three studies: part 2. The Women's Health Initiative: estrogen plus progestogen. AB - BACKGROUND: Based principally on findings in three studies, the Collaborative Reanalysis (CR), the Women's Health Initiative (WHI), and the Million Women Study (MWS), it is claimed that combined hormone replacement therapy (HRT) with estrogen plus progestogen is now an established cause of breast cancer. For unopposed estrogen therapy the evidence in the three studies is conflicting: the CR and MWS have reported increased risks in estrogen users, while the WHI has not. The authors have previously reviewed the findings in the CR (Part 1). OBJECTIVE: To evaluate the evidence for causality in the WHI studies. METHODS: Using generally accepted causal criteria, in this paper (Part 2) the authors evaluate the findings in the WHI for estrogen plus progestogen; in a related paper (Part 3) the authors evaluate the findings for unopposed estrogen. An evaluation of the MWS (Part 4), and of trends in breast cancer incidence following publication of the WHI findings in 2002 (Part 5) will follow. RESULTS: For estrogen plus progestogen the findings did not adequately satisfy the criteria of bias, confounding, statistical stability and strength of association, duration-response, internal consistency, external consistency or biological plausibility. CONCLUSION: HRT with estrogen plus progestogen may or may not increase the risk of breast cancer, but the WHI did not establish that it does. PMID- 21642265 TI - Effects of resistance training in children and adolescents: a meta-analysis. PMID- 21642266 TI - Ethical practice and sports physician protection: a proposal. PMID- 21642267 TI - Physiotherapists administering medications under instruction. PMID- 21642268 TI - Medicolegal neglect? The case for physical activity promotion and exercise medicine. PMID- 21642269 TI - Evaluation of Green Dot: an active bystander intervention to reduce sexual violence on college campuses. AB - Using a cross-sectional survey of a random sample of 7,945 college undergraduates, we report on the association between having received Green Dot active bystander behavior training and the frequency of actual and observed self reported active bystander behaviors as well as violence acceptance norms. Of 2,504 students aged 18 to 26 who completed the survey, 46% had heard a Green Dot speech on campus, and 14% had received active bystander training during the past 2 years. Trained students had significantly lower rape myth acceptance scores than did students with no training. Trained students also reported engaging in significantly more bystander behaviors and observing more self-reported active bystander behaviors when compared with nontrained students. When comparing self reported active bystander behavior scores of students trained with students hearing a Green Dot speech alone, the training was associated with significantly higher active bystander behavior scores. Those receiving bystander training appeared to report more active bystander behaviors than those simply hearing a Green Dot speech, and both intervention groups reported more observed and active bystander behaviors than nonexposed students. PMID- 21642270 TI - Which urgent care services do febrile children use and why? AB - OBJECTIVES: To explore how parents navigate urgent and emergency care (U&EC) services when their child <5 years old has a feverish illness, their views of that experience and whether services are meeting their needs and triaging in line with national guidance. DESIGN: Parents of a febrile child <5 years old contacting any U&EC service in three localities during a 6-month period were invited to participate in a telephone questionnaire supplemented by case note review. A subset participated in an in-depth interview. RESULTS: Of 556 parents expressing an interest, 220 enrolled, making 570 contacts (median 3, range 1-13) with services during the child's illness which lasted 3 days on average. Parents' first preference for advice in hours was to see a general practitioner (GP) (67%; 93/138) and when unavailable, National Health Service Direct (46%; 38/82). 155 made more than one contact and 63% of the repeat contacts were initiated by a service provider. A range of factors influenced parents' use of services. Parents who reported receiving 'safety netting' advice (81%) were less likely to re present to U&EC services than those who did not recall receiving such advice (35% vs 52%, p=0.01). Parents identified a need for accurate, consistent, written advice regarding fever and antipyretics. CONCLUSION: Parents know the U&EC service options available, and their first choice is the GP. Multiple contacts are being made for relatively well children, often due to repeated referrals within the system. Safety netting advice reduces re-attendances but parents want explicit and consistent advice for appropriate home management. PMID- 21642271 TI - Policy and pragmatism in breast feeding. PMID- 21642272 TI - Homeopathic medicines for children. AB - This article describes the homeopathic tradition and considers the safety, manufacture, effectiveness and regulation of homeopathic medicines. These medicines are commonly purchased without prescription for children, so an understanding of the basis of therapy is important to ensure appropriate and safe usage. The role of integrated medicine in the National Health Service is also reviewed with identification of research priorities. PMID- 21642273 TI - Prospective study on central venous line associated bloodstream infections. AB - OBJECTIVE: To prospectively assess the incidence rates and characteristics of central venous line associated bloodstream infections (CLABSI) in one institution. METHODS: All patients with indwelling central venous catheters (CVC) between 1 April 2008 and 31 March 2009 were enrolled. The medical records of patients were reviewed and information on relevant characteristics entered into a standardised questionnaire. Central laboratory records were regularly checked for positive blood cultures in study patients. RESULTS: There were 209 CVC for a total of 14752 CVC days in 152 patients (88 males, 58%) including neonates and patients with surgical conditions and haemato-oncological and other underlying diseases. Median age at CVC insertion was 3 months (IQR 0-56 months). Fourteen CLABSI occurred in 13 patients. Overall CLABSI incidence (per 1000 CVC days) was 0.95 (9.71 for silastic percutaneous CVC, 7.65 for other CVC, 1.97 for Broviac, 0.18 for Port-a-cath). CVC remained in place for <= 14 days in 109 (52%) instances, 15-90 days in 45 (22%) instances and > 90 days in 55 (26%) instances. The incidence of CLABSI in these three categories was 3.36, 4.36 and 0.47, respectively. Predominating cultured organisms were coagulase-negative staphylococci (N=3), S. aureus (N=3), and Enterococcus spp. (N=3). CONCLUSIONS: CLABSI incidence varied by type of catheter and type of patient, with the highest risk in neonates (with silastic percutaneous CVC) and by far the lowest risk for Port-a-cath CVC. Prophylactic measures to reduce CLABSI should be tailored to individual types of catheters and patient characteristics. PMID- 21642274 TI - GhHyPRP4, a cotton gene encoding putative hybrid proline-rich protein, is preferentially expressed in leaves and involved in plant response to cold stress. AB - Plant hybrid proline-rich proteins (HyPRPs) usually consist of an N-terminal signal peptide, a central proline-rich domain, and a conserved eight-cysteine motif C-terminal domain. In this study, one gene (designated as GhHyPRP4) encoding putative HyPRP was isolated from cotton cDNA library. Northern blot and quantitative reverse transcriptase-polymerase chain reaction analyses revealed that GhHyPRP4 was preferentially expressed in leaves. Under cold stress, GhHyPRP4 expression was significantly up-regulated in leaves of cotton seedlings. Using the genome walking approach, a promoter fragment of GhHyPRP4 gene was isolated from cotton genome. GUS (beta-glucuronidase) gene driven by GhHyPRP4 promoter was specifically expressed in leaves and cotyledons of the transgenic Arabidopsis thaliana. Furthermore, GUS expression in leaves was remarkably induced by cold stress. Overexpression of GhHyPRP4 in yeast (Schizosaccharomyces pombe) significantly enhanced the cell survival rate upon treatment under -20 degrees C for 60 h. These data suggested that GhHyPRP4 may be involved in plant response to cold stress during seedling development of cotton. PMID- 21642275 TI - Cucurbitacin B induces rapid depletion of the G-actin pool through reactive oxygen species-dependent actin aggregation in melanoma cells. AB - Cucurbitacin B (CuB), a triterpenoid compound isolated from Cucurbitaceae plants, has been reported as a promising anti-cancer agent, yet its action mechanism is still controversial. In this study, we explored the potential mechanism of CuB in murine B16F10 melanoma cells. Anti-proliferation and anti-invasion effects were assessed in cultured cells, and in vivo anti-tumor activity was evaluated in a murine subcutaneous melanoma model. Flow cytometry was adopted to analyze cell cycle distribution and reactive oxygen species (ROS) levels. Actin levels were determined by western blot analysis, and the profiles of differential expressed proteins were identified by a quantitative proteomic approach. The results showed that CuB exerted inhibitory effects on cell proliferation, colony formation, as well as migration and invasion potential of the melanoma cells. The growth of subcutaneous melanoma was significantly inhibited in mice treated with CuB when compared with control group. Furthermore, CuB treatment caused rapid cell membrane blebbing and deformation, and induced G(2)/M-phase arrest and formation of multiploid cells. Notably, the G-actin pool was rapidly depleted and actin aggregates were formed quickly after CuB treatment. A number of cytoskeleton regulatory proteins were differentially regulated. Blockage of ROS production significantly reduced the G-actin depletion ability and the anti-tumor activity of CuB. These findings indicate that CuB induces rapid depletion of the G-actin pool through ROS-dependent actin aggregation in melanoma cells, which may at least partly account for its anti-tumor activity. PMID- 21642276 TI - mTOR and the differentiation of mesenchymal stem cells. AB - The mammalian target of rapamycin (mTOR), an evolutionarily conserved serine threonine protein kinase, belongs to the phosphoinositide 3-kinase (PI3K)-related kinase family, which contains a lipid kinase-like domain within their C-terminal region. Recent studies have revealed that mTOR as a critical intracellular molecule can sense the extracellular energy status and regulate the cell growth and proliferation in a variety of cells and tissues. This review summarizes our current understanding about the effects of mTOR on cell differentiation and tissue development, with an emphasis on the lineage determination of mesenchymal stem cells. mTOR can promote adipogenesis in white adipocytes, brown adipocytes, and muscle satellite cells, while rapamycin inhibits the adipogenic function of mTOR. mTOR signaling may function to affect osteoblast proliferation and differentiation, however, rapamycin has been reported to either inhibit or promote osteogenesis. Although the precise mechanism remains unclear, mTOR is indispensable for myogenesis. Depending on the cell type, rapamycin has been reported to inhibit, promote, or have no effect on myogenesis. PMID- 21642277 TI - Clinical factors associated with the non-utilization of an anaesthesia incident reporting system. AB - BACKGROUND: Incident reporting is a widely recommended method to measure undesirable events in anaesthesia. Under-utilization is a major weakness of voluntary incident reporting systems. Little is known about factors influencing reporting practices, particularly the clinical environment, anaesthesia team composition, severity of the incident, and perceived risk of litigation. The purpose of this study was to assess each of these, using an existing anaesthesia database. METHODS: We performed a retrospective cohort study and analysed 46 207 surgical patients. We used multivariate analysis to identify factors associated with the non-utilization of the reporting system. RESULTS: We found that in 7022 (15.1%) of the procedures performed, the incident reporting system was not used. Factors associated with the non-use of the system were regional anaesthesia/local anaesthesia, odds ratio (OR) 1.64 [95% confidence interval (CI) 1.03-2.62], emergency procedures OR 1.15 (95% CI: 1.05-1.27), and a consultant anaesthetist working without a trainee, OR 1.71 (95% CI: 1.03-2.82). In contrast, factors such as longer duration of surgery, OR 0.85 (95% CI: 0.76-0.94), the presence of a senior anaesthesia trainee, OR 0.86 (95% CI: 0.81-0.92), and the occurrence of severe complications with a high risk of litigation (i.e. death, nerve injuries) were less associated with a non-use of the reporting system, OR 0.65 (95% CI: 0.44-0.97). Team composition and time of day had no measurable impact on reporting practices. CONCLUSIONS: Clinical factors play a significant role in the utilization of an anaesthesia incident reporting system and more particularly, severity of complications and higher liability risks which appear more as incentives than barriers to incident reporting. PMID- 21642278 TI - Acute coronary syndrome and endocarditis 20 years before: how do they match? PMID- 21642279 TI - Invasion of lung mesenchymal chondrosarcoma into the left atrium via the pulmonary vein detected on transoesophageal echocardiography. PMID- 21642280 TI - Interventricular septum haematoma with right ventricle obstruction caused by percutaneous coronary intervention: case report. PMID- 21642281 TI - Mitral annulus and left atrium wall abscess fistulized to the left ventricle. PMID- 21642282 TI - Cardiac bone marrow cell therapy: the proof of the pudding remains in the eating. PMID- 21642283 TI - Clinical course and outcome of patients enrolled in US and non-US centres in MADIT-CRT. AB - AIMS: We aimed to evaluate within the MADIT-CRT database whether different enrollment characteristics between US and non-US centres affected the clinical course of study patients. METHODS AND RESULTS: We evaluated differences in baseline characteristics, procedure-associated complications, clinical as well as echocardiographic response to cardiac resynchronization therapy with a defibrillator (CRT-D), between patients enrolled in 87 US centres (n = 1271) and 23 non-US centres (n = 549) in MADIT-CRT. Non-US patients displayed significant differences in baseline characteristics from US patients, including a higher frequency of left bundle branch block, a more advanced heart failure (HF) functional class >3 months prior to enrollment, and larger baseline cardiac volumes. Procedure-related complications occurred at a significantly higher frequency among patients enrolled in non-US centres (17%) than among those enrolled in US centres (10%; P < 0.001). During follow-up, CRT-D was associated with 42% (P = 0.003) and 38% (P < 0.001) reductions in the risk of HF or death in the two respective groups (P for the difference = 0.80), and with similar reductions in cardiac volumes (all P > 0.10). Subgroup analysis showed a more pronounced effect of CRT-D among women in the US group, including a significant 71% (P = 0.02) reduction in the risk of death, whereas CRT-D therapy was associated with a significant clinical benefit in men only in the non-US group. CONCLUSION: Patients enrolled in US and non-US centres in MADIT-CRT displayed significant differences in baseline clinical and echocardiographic characteristics and in the frequency of procedure-related complications, but experienced an overall similar clinical and echocardiographic response to CRT-D. PMID- 21642285 TI - Macrophage infiltration and smooth muscle cells content associated with haptoglobin genotype in human atherosclerotic carotid plaques. AB - We assessed the association between the haptoglobin (Hp) genotype and 2 common indicators of atherosclerotic plaque instability: macrophage infiltration and the smooth muscle cell (SMC) content. A total of 70 consecutive patients who underwent carotid endarterectomy were included in the study. For immunohistochemical study the anti-CD68 and anti-a-actin antibodies were used on adjacent serial sections; 36 plaques from patients with the Hp 1-1 or 2-1 genotype and 34 plaques from patients with the Hp 2-2 genotype were analyzed. The macrophage content (CD68+) was significantly higher in the Hp 2-2 group compared with that in the Hp 1-1 or 2-1 group (P < .001). In plaques from patients with diabetes, the SMC content was significantly lower in the Hp 2-2 group (P = .034). Carotid plaques from diabetic patients with Hp 2-2 genotype had higher macrophage infiltration and lower SMC content. Both parameters are indicators of atherosclerotic plaque instability. PMID- 21642286 TI - The influence of risk factors for metabolic syndrome on vascular complications. AB - We assessed the influence of metabolic syndrome (MetS) characteristics on vascular complications. The study included 108 patients (72 women and 36 men, age 57.0 +/- 8.5 years) with MetS that was diagnosed according to International Diabetes Federation criteria. The prevalence of micro- and macrovascular complications was assessed: vascular changes in the fundus of the eye-72%, ischemic heart disease-54.9%, estimated glomerular filtration rate (eGFR) <90 mL/min-38.9%, diabetic foot-5.55%, and cerebrovascular accident-3.7% of patients. A negative correlation between high-density lipoprotein cholesterol (HDL-C) concentration, creatinine levels, and level of vascular changes in the fundus of the eye was found. Moreover, the level of obesity and fasting glucose level had positive correlation with the intensity of vascular changes in the fundus of the eye. Characteristics of MetS (obesity and fasting glucose level) have high impact on advancing vascular complications in these patients. High level of HDL-C decreases the intensity of vascular changes in patients with MetS. PMID- 21642284 TI - A high-throughput immobilized bead screen for stable proteins and multi-protein complexes. AB - We describe an in vitro colony screen to identify Escherichia coli expressing soluble proteins and stable, assembled multiprotein complexes. Proteins with an N terminal 6His tag and C-terminal green fluorescent protein (GFP) S11 tag are fluorescently labeled in cells by complementation with a coexpressed GFP 1-10 fragment. After partial colony lysis, the fluorescent soluble proteins or complexes diffuse through a supporting filtration membrane and are captured on Talon((r)) resin metal affinity beads immobilized in agarose. Images of the fluorescent colonies convey total expression and the level of fluorescence bound to the beads indicates how much protein is soluble. Both pieces of information can be used together when selecting clones. After the assay, colonies can be picked and propagated, eliminating the need to make replica plates. We used the method to screen a DNA fragment library of the human protein p85 and preferentially obtained clones expressing the full-length 'breakpoint cluster region-homology' and NSH2 domains. The assay also distinguished clones expressing stable multi-protein complexes from those that are unstable due to missing subunits. Clones expressing stable, intact heterotrimeric E.coli YheNML complexes were readily identified in libraries dominated by complexes of YheML missing the N subunit. PMID- 21642287 TI - Low estimated glomerular filtration rate is a major determinant of low ankle brachial index and toe-brachial index in type 2 diabetes. AB - We enrolled 1461 Taiwanese type 2 diabetic outpatients with ankle-brachial index (ABI) and toe-brachial index (TBI) examinations, excluding participants with history of stroke, end-stage renal disease, malignancy, acute myocardial infarction, amputation, and overt calcification of the lower limbs (ABI > 1.3). Ankle-brachial index values <0.9 were found in 2.8% of the patients and 5.7% had TBI < 0.6. Estimated glomerular filtration rate (eGFR; 90 +/- 33 mL/min per 1.73 m2) obtained from 473 patients correlated significantly with both ABI and TBI. Progressive eGFR decline was observed in 419 participants with normal ABI and TBI, 35 with normal ABI but low TBI, and 19 with low ABI and normal or low TBI (P for trend < .001). After adjusting for confounding factors, age and eGFR were significantly associated with TBI and ABI. Low eGFR is associated with peripheral arterial disease in type 2 diabetes with mild to moderate renal insufficiency. PMID- 21642288 TI - The role of macrophage colony-stimulating factor in patients with acute myocardial infarction: a pilot study. AB - We assessed whether macrophage colony-stimulating factor (M-CSF) levels are associated with left ventricular systolic dysfunction (LVSD) in patients with acute myocardial infarction (AMI). We studied 56 patients with AMI (mean age: 67 +/- 12 years) and identified those with clinical (Killip class >II) or echocardiographic signs (ejection fraction <=45%) of LVSD. We evaluated the established cardiovascular risk factors and measured several cardiovascular biomarkers, including M-CSF. Serum M-CSF concentrations (pg/mL) were significantly increased in patients with both clinical and echocardiographic signs of LVSD (460 +/- 265 vs 290 +/- 210, P = .0103 and 493 +/- 299 vs 287 +/- 174, P = .0028, respectively). We found a significant inverse association between M-CSF and ejection fraction (r = -.351, P = .0079). Logistic regression analysis revealed that, among all evaluated clinical and biochemical parameters, the stronger predictor of LVSD was M-CSF (odds ratios 2.1, 95% confidence interval 1.1-2.9, P = .0168). This is the first study reporting plasma M-CSF levels as independent determinants of low LV ejection fraction and clinical LV dysfunction in patients with AMI. PMID- 21642289 TI - Asymptomatic carotid artery disease in valvular heart surgery: impact of systematic screening on surgical strategy and neurological outcome. AB - We evaluated the prevalence of asymptomatic carotid artery disease in patients scheduled for valvular cardiac surgery. Preoperative screening of the carotid arteries was performed. Among 1012 patients scheduled for valvular cardiac surgery, 267 (26.4%) had carotid stenosis graded >50%; 37 had carotid stenosis >70% and underwent combined valvular surgery and carotid endarterectomy (CEA); and 230 (86%) had carotid stenosis >50% to <= 69% and received valvular cardiac surgery under hypothermic cardiopulmonary bypass. Operative mortality and the rate of perioperative adverse neurological events were comparable among the groups. During 6.8 years of follow-up, patients with carotid stenosis not exceeding 69% at the time of surgery had CEA more frequently (P < .05) and stroke/transient ischemic attack ([TIA] P < .05) versus patients treated with combined surgery. The prevalence of asymptomatic carotid stenosis is not negligible in patients undergoing isolated valvular surgery. Combined valvular and carotid surgery is safe and reduces the incidence of CEA and stroke/TIA during follow-up. PMID- 21642290 TI - Low rate of fluoroquinolone resistance in Mycobacterium tuberculosis isolates from northern Tanzania. AB - OBJECTIVES: Fluoroquinolones are used in second-line treatment of tuberculosis (TB) and have a potential role in shortening TB treatment duration. The wide use of fluoroquinolones in the treatment of other infections, including respiratory tract infections in patients with (undiagnosed) active TB, could result in fluoroquinolone-resistant Mycobacterium tuberculosis. We determined the rate of fluoroquinolone resistance in M. tuberculosis isolates obtained from Tanzanian patients and linked this to previous fluoroquinolone exposure and mycobacterial resistance to rifampicin and isoniazid. METHODS: A total of 291 M. tuberculosis isolates were obtained between April 2009 and June 2010 from patients with smear positive pulmonary TB and tested for susceptibility to ciprofloxacin, moxifloxacin, rifampicin and isoniazid. Information on previous fluoroquinolone use was obtained by interviewing patients and checking their medical files. RESULTS: Only 2 (0.7%) of the 291 M. tuberculosis isolates were resistant to ciprofloxacin; 1 of which was intermediately resistant to moxifloxacin as well. These two isolates were susceptible to rifampicin and isoniazid. Twenty-two (8%) of the 291 patients had a history of fluoroquinolone use (median: 7 days; interquartile range: 5-10 days). The patients from whom the fluoroquinolone resistant M. tuberculosis isolates were obtained had no known history of previous fluoroquinolone use. CONCLUSIONS: Our findings indicate that the rate of fluoroquinolone-resistant M. tuberculosis in Tanzanian patients with TB is low and not related to previous, brief episodes of exposure to fluoroquinolones. The findings favour future application of fluoroquinolones in TB treatment regimens of shorter duration. PMID- 21642291 TI - Multidrug-resistant tuberculosis (MDR-TB) treatment in the UK: a study of injectable use and toxicity in practice. AB - BACKGROUND: Multidrug-resistant tuberculosis (MDR-TB) is an increasing challenge to health services globally. Although new drugs are in development, current guidelines still recommend prolonged use of injectable antimicrobials (usually amikacin, kanamycin or capreomycin). The evidence base to inform treatment and monitoring strategies is very limited. METHODS: We conducted a retrospective study of patients initiating injectable antimicrobials for MDR-TB treatment in five UK centres between January 2004 and December 2009. (i) Current treatment and monitoring strategies were reviewed. (ii) The incidence of ototoxicity (defined both clinically and on audiological testing) and factors associated with ototoxicity were investigated using logistic regression. RESULTS: (i) The choice of injectable antimicrobial varied. Of 50 MDR-TB patients, 29/50 (58%) received amikacin, 11/50 (22%) received capreomycin and 10/50 (20%) received streptomycin or a combination; reflecting a difference in policy between centres. Only 21/50 (42%) patients received baseline screening by audiogram within 2 weeks of starting treatment and 16/50 (32%) then had monthly audiograms, with the majority screened more infrequently and 12/50 (24%) receiving no screening. (ii) Of the 50 patients, 14 (28%) experienced ototoxicity, with 9/50 (18%) left with long-term hearing loss. Increased age (P = 0.02), use of amikacin (P = 0.02) and decreased renal function (P = 0.01) were significantly associated with ototoxicity. CONCLUSIONS: There is local variation in both the choice of injectable agent and in ototoxicity screening practices. Long-term morbidity from injectable treatment is significant even in this well-resourced setting, and the data suggest capreomycin might be associated with less ototoxicity when compared with amikacin. There is a need for more high-quality clinical data to inform future guidelines for treatment and monitoring. PMID- 21642292 TI - Pulmonary resection for patients with multidrug-resistant tuberculosis: systematic review and meta-analysis. AB - OBJECTIVES: Multidrug-resistant tuberculosis (MDR-TB) has become an emerging global public health crisis. Several studies have suggested that pulmonary resection has efficacy in the treatment of MDR-TB. A systematic review of the available therapeutic studies was conducted to determine the treatment outcome among patients with MDR-TB who underwent pulmonary resection. METHODS: To evaluate pulmonary resection for MDR-TB, a random-effect meta-analysis of the available studies was used to assess the overall treatment outcome. Subgroup analyses were also conducted by separating studies based on each characteristic independently. RESULTS: After screening 4996 articles, 15 clinical reports with a mean of 63 patients per report met the inclusion criteria. Analysis of the studies showed that the estimated pooled treatment success rate of pulmonary resection for patients with MDR-TB was 84% [95% confidence interval (CI) 78% 89%]. The rates of failure, relapse, death and default were 6% (95% CI 4%-8%), 3% (95% CI 1%-4%), 5% (95% CI 2%-8%) and 3% (95% CI 1%-5%), respectively. The proportion of patients treated successfully did not differ significantly on the basis of any of the individual study characteristics. CONCLUSIONS: Substantial heterogeneity in the study characteristics prevented a more conclusive determination of what factors had the greatest effect on the proportion of patients that achieve treatment success and limited the validity of this analysis. Some important variables, including patient HIV status, were inconsistently reported between studies. These results underscore the importance of strong patient support and treatment follow-up systems to develop successful MDR-TB treatment programmes. PMID- 21642293 TI - Clinical and prognostic effects of atrial fibrillation in heart failure patients with reduced and preserved left ventricular ejection fraction. AB - AIMS: Atrial fibrillation (AF) is common in heart failure (HF), but few data regarding the prognostic relevance of AF are available in HF patients with preserved left ventricular ejection fraction (HF-PEF). We aimed to study the clinical impact of AF vs. sinus rhythm (SR) in stabilized HF patients with reduced left ventricular ejection fraction (HF-REF) and in those with preserved left ventricular ejection fraction (HF-PEF). METHODS AND RESULTS: We studied 927 patients with stable HF, of whom 336 (36%) had AF. N-terminal pro-B-type natriuretic peptide (NT-proBNP) concentrations were measured at baseline and patients were followed for 18 months. We compared time to first HF (re )hospitalization or death between patients with AF and SR. Atrial fibrillation was present at baseline in 215 (35%) patients with HF-REF (mean LVEF 0.25 + 0.08) and in 121 (40%) patients with HF-PEF (mean LVEF 0.50 + 0.09). Plasma NT-proBNP levels were similar in AF and SR patients (median 2398 vs. 2532 pg/mL, P = 0.74). Atrial fibrillation was independently associated with elevated NT-proBNP levels in HF-PEF, but not in HF-REF patients (multivariable B = 0.33, P= 0.047 and B = 0.03; P = 0.89, respectively). After 18 months of follow-up, the presence of AF was an independent predictor of death or HF hospitalization in HF-PEF (multivariable hazard ratio 1.49 (95% CI 1.04-2.14), P = 0.03), but not in HF-REF patients (1.05 (CI 95% 0.80-1.38), P = 0.72). CONCLUSION: Atrial fibrillation is equally common in patients with HF-PEF and HF-REF. In HF-PEF, but not in HF-REF patients, AF was associated with higher NT-proBNP levels and was independently related to death or HF hospitalization. PMID- 21642294 TI - Genetic association studies of angiogenesis- and vasoconstriction-related genes in women with recurrent pregnancy loss: a systematic review and meta-analysis. AB - BACKGROUND: Angiogenesis and an adequate blood supply are critical for several steps in human early pregnancy. Some studies have reported angiogenesis- and vasoconstriction-related genes are associated with recurrent pregnancy loss (RPL), but their sample size was limited. This study was conducted to investigate the genetic association between these angiogenesis- and vasoconstriction-related genes and idiopathic RPL, using meta-analyses. METHODS: A systematic review of the published literature from MEDLINE and EMBASE databases was conducted and investigations of an angiogenesis- and vasoconstriction-related gene polymorphism in RPL reported more than three times were selected. Aggregating data from eligible studies were integrated into meta-analyses by means of random effects models. RESULTS: Of 185 potentially relevant studies, 18 case-control studies comprising a total of 2397 RPL patients and 1760 controls were included into the meta-analyses. Among these genetic association studies were 4 reports of vascular endothelial growth factor (VEGF) (-1154G>A) polymorphisms, 4 reports of p53 (codon72) and 10 reports of endothelial nitric oxide synthase (eNOS) (B/A, Glu298Asp) with RPL. The integrated results showed that VEGF (-1154G>A), p53 (codon 72) and eNOS (Glu298Asp) polymorphisms were significantly associated with RPL, and their summary odd ratios [95% confidence interval (CI)] were 1.51 (1.13 2.03), 1.84(1.07-3.16) and 1.37 (1.11-1.69), respectively. The summary odd ratio of the eNOS (B/A) polymorphism in RPL was 1.15 (0.94-1.41), and failed to show significance at meta-analysis. CONCLUSIONS: Meta-analyses of available data showed significant associations between the VEGF (-1154G>A), p53 (codon72) and eNOS (Glu298Asp) polymorphisms and idiopathic RPL. These angiogenesis- and vasoconstriction-related genes jointly confer higher susceptibility to idiopathic RPL. PMID- 21642295 TI - Severe Taenia ovis outbreak in a sheep flock in south-west England. PMID- 21642296 TI - Perceptions of clients and veterinarians on what attributes constitute 'a good vet'. AB - The perceptions of veterinarians and small animal (SA) clients on what attributes constitute 'a good veterinarian' were examined by a questionnaire survey. The respondents were asked to record how important they considered 20 attributes for a veterinary surgeon to have on a five-point scale from 'not at all important' to 'very important'. In addition, they were asked to list which attributes they considered to be the three most important attributes in a veterinary surgeon; finally, they were asked whether there were any additional attributes that they considered to be highly desirable in a veterinary surgeon. In total, 407 SA clients, 243 SA veterinarians and 61 non-SA veterinarians completed the questionnaire. There were significant differences in the proportion of clients who considered an attribute to be 'very important' compared with SA veterinarians for 12 of the 20 attributes (P<0.005). A larger proportion of clients considered 'confidence', 'knowledge about veterinary medicine and surgery', 'cleanliness', 'good at explaining technical terms', 'patience', 'clear about cost of treatment', 'ability to work in a team', 'honesty', 'politeness', 'decisiveness', 'good with animals' and 'good practical skills' to be 'very important' attributes than the SA veterinarians; a larger proportion of SA veterinarians considered 'good communication skills' to be a 'very important' attribute than the clients. PMID- 21642297 TI - Long-term efficacy and safety of firocoxib in the treatment of dogs with osteoarthritis. AB - Thirty-nine client-owned dogs with osteoarthritis were treated with 5 mg/kg firocoxib administered orally, once a day for 52 weeks. Veterinary examinations were performed on approximately days 0, 15, 90, 180, 270 and 360. Twenty-five dogs completed the study. The withdrawal rate associated with gastrointestinal side effects was low (5.1 per cent of dogs). Based on the owners' assessment, 82 per cent of the dogs had improved at day 15, 84 per cent of the 32 remaining dogs had improved at day 90, and 96 per cent of the 25 dogs that completed the trial had improved at day 360. During this trial, 12 (48 per cent) of the 25 remaining dogs showed an improvement in their lameness from day 90 to day 360 (P<0.05). PMID- 21642299 TI - OIE declares rinderpest eradicated. PMID- 21642301 TI - Selecting the best mate online. PMID- 21642302 TI - Exploring the wider health benefits of companion animals. PMID- 21642303 TI - Disciplinary Committee dismisses fraudulent registration charges. PMID- 21642305 TI - Vets asked to raise awareness of pesticide risks to pets. PMID- 21642306 TI - Managing crises in the food chain. PMID- 21642309 TI - Dealing with a potential case of FCV-associated virulent systemic disease. PMID- 21642310 TI - Gender balance in the veterinary profession. PMID- 21642311 TI - Water bird mortality associated with freezing winter weather and disease. PMID- 21642312 TI - Preventing dog bite wounds in people. PMID- 21642320 TI - Relationships between body mass index, cardiovascular mortality, and risk factors: a report from the SCORE investigators. AB - BACKGROUND: Although cardiovascular disease (CVD) is the biggest global cause of death, CVD mortality is falling in developed countries. There is concern that this trend may be offset by increasing levels of obesity. DESIGN: We used the Systematic Coronary Risk Evaluation (SCORE) data set to examine relationships between body mass index (BMI), conventional risk factors and CVD mortality. METHODS: The SCORE data set comprises data from 12 European cohort studies. The relationship between BMI and CVD mortality was examined in each BMI category using univariable and multivariable (Cox) analyses. The SCORE population was also divided into gender and age strata: under 40, 40-49, 50-59, and over 60. The rate of CVD mortality in each BMI category was calculated within each gender and age stratum. Relationships between BMI and other CVD risk factors were also examined. RESULTS: There was a strong, graded but J-shaped univariable relationship between BMI and CVD mortality in both genders. Each 5-unit increase in BMI was associated with an increase in CVD mortality of 34% in men and 29% in women. The hazard ratios remained significant when adjusted for age, self-reported smoking status, total cholesterol, and systolic blood pressure (SBP). On additional adjustment for diabetes and high-density lipoprotein cholesterol (HDL), the association between BMI and CVD mortality did not persist. In all age groups except those over 60 there were significant relationships between increased BMI and CVD mortality. In the over-60 age group the only significant relationships with mortality were in underweight and severely overweight women and mildly obese men. After adjustment for age, each 1-unit increase in BMI was associated with a 1.14 mmHg increase in SBP, 0.055 mmol/l increase in total cholesterol, and a 0.024 mmol/l decrease in HDL in men. Figures were slightly lower in women. CONCLUSIONS: Overall, overweight and obesity relate to CVD mortality in a strong and graded manner. The effects are greater in women and markedly so in younger persons. It is likely that a substantial part of the BMI-associated risk of CVD mortality is mediated through other known CVD risk factors. This increases the public health importance of BMI as both a simple indicator and mediator of CVD risk. PMID- 21642321 TI - Specialized cardiological care may be overutilized in urban areas of Quebec. AB - BACKGROUND: Urban/rural differences in secondary cardiovascular disease (CVD) events have previously been observed for Quebec. These differences could be attributable to differential utilization of specialized cardiological care, such as revascularization procedures and visits to cardiologists; if this were the case, policies to increase utilization in rural areas would be indicated. DESIGN: This is a population-based cohort study. METHODS: We analysed mortality and hospital re-admission in Quebec within 1 year after an initial cardiovascular event in relation to urban/rural location and specialized care utilization, controlling for demography, comorbidities, and cumulative hospitalization. RESULTS: Analysis showed higher hospital re-admissions and slightly lower CVD mortality in rural areas, as well as less use of specialized care in rural areas. However, urban/rural differentials were not attributable to differences in utilization of care. Paradoxically, comorbidities were lower among patients who saw specialists. CONCLUSIONS: Ultimately, urban/rural differences in secondary CVD outcomes were not attributable to differences in care utilization or our measures of underlying health status, and were likely due to cultural or lifestyle factors that are both hard to model and hard to change through policy. There appears to be overutilization of specialized care in urban areas, an issue which requires further study. Our results suggest that substantial caution is required when interpreting health service usage data and that critical factors in the relationship between specialized cardiological care and outcomes are still poorly understood at a population level. PMID- 21642322 TI - Knowledge, attitude and practice of home management of childhood diarrhoea among caregivers of under-5 children with diarrhoeal disease in Northwestern Nigeria. AB - OBJECTIVES: Diarrhoeal diseases (DDs) constitute public health problem. Reduction in related mortality and morbidity hinges on active participation of home caregivers. It is pertinent to determine the degree of their empowerment. DESIGN: Cross-sectional study. SETTING: Randomly selected community primary health centres in northwestern Nigerian. SUBJECTS: Home caregivers presenting with children aged 1-59 months having DD. OUTCOME MEASURES: Knowledge, attitude and practice regards home management of DD. RESULTS: Less than 1% of caregivers was knowledgeable about home management of DD. Antibiotics and anti-diarrhoeal agents use was common at 36%. ORS use was abysmally low at 8.6%. Only 32% of caregivers were aware of the use of zinc in the management of DD. Adherence to 10-day zinc supplementation was encouraging at 75.5%. CONCLUSION: There is an urgent need to scale up educational and promotional activities with regards to the home management of DD if the millennium development goals are to be met. PMID- 21642323 TI - Diagnosis and management of premenstrual disorders. PMID- 21642324 TI - Minister orders new checks on private care homes after regulator failed to act on abuse. PMID- 21642325 TI - WHO expert group classifies mobile phones as "possibly carcinogenic" to humans. PMID- 21642326 TI - Give GPs more control over care planning, says Audit Scotland. PMID- 21642327 TI - Poland violated human rights of woman who was refused timely access to prenatal tests. PMID- 21642328 TI - Governments in sub-Saharan Africa need to tackle chronic disease burden. PMID- 21642329 TI - GPs are "raring to go" with commissioning of health care, alliance claims. PMID- 21642330 TI - UK prime minister "guarantees" welfare of residents of struggling nursing home chain. PMID- 21642331 TI - Commentary on 'Opt-out HIV testing: an ethical analysis of women's reproductive rights'. PMID- 21642332 TI - A problem with inclusion in learning disability research. AB - People with severe learning disability are particularly difficult to include in the research process. As a result, researchers may be tempted to focus on those with learning disability who can be included. The problem is exacerbated in this field as the political agenda of inclusion and involvement is driven by those people with learning disability who are the higher functioning. To overcome this we should first detach the notion of consent from ideas about autonomy and think instead of it as a way to avoid wronging others; this fits the original historical use of consent in research. This allows us to think in terms of including participants to the best of their abilities rather than in terms of a threshold of autonomy. Researchers could then use imaginative ways to include the least able and to ensure they are not wronged in research or by exclusion from it. PMID- 21642333 TI - Compassion in care: a qualitative study of older people with a chronic disease and nurses. AB - This article describes compassion as perceived within the relationship between nurses and older persons with a chronic disease. The aim of the study is to understand the benefit of compassion for nursing practice within the context of long-term care. The design of the study involves a qualitative analysis of in depth interviews with nurses and patients in three different care-settings. Results show the nature of compassion in seven dimensions: attentiveness, listening, confronting, involvement, helping, presence and understanding. Analysis of the data also shows in what way opinions of participants relate to issues raised in a previous literature study, for example the difference between pity and compassion. The conclusion states that compassion is a valuable process which motivates patients as well as nurses to cooperate in achieving relevant outcomes of care. The discussion involves some methodological issues. For one thing, further confirmation of the dimensions found is recommended. PMID- 21642334 TI - Opt-out HIV testing: an ethical analysis of women's reproductive rights. AB - As the HIV epidemic continues to grow worldwide, women are increasingly and disproportionally affected. With the introduction of anti-retroviral medications that have been found to effectively prevent perinatal transmission of HIV, the approach to HIV testing in pregnant women has grown increasingly more controversial. In recent years, the model of voluntary counseling and testing (VCT) has come into question with opt-out testing now advocated for by the Centers for Disease Control and occurring widely in pregnancy. The benefits of opt-out testing are numerous and may justify its use in replacing the VCT that many have come to see as insufficient. An ethical analysis of opt-out testing suggests it may be at odds with true informed consent and involve a degree of coercion that would not be allowed outside the prenatal setting. If opt-out testing is going to remain the standard of care then the ethical issues it raises must be made transparent. Strategies need to be designed for ensuring that HIV counseling and testing in pregnancy is done in accordance with ethical and reproductive rights principles. PMID- 21642335 TI - Nasogastric feeding at the end of life: a virtue ethics approach. AB - The use of Nasogastric (NG) feeding in the provision of artificial nutrition and hydration at the end of life has, for the most part, been regarded as futile by the medical community. This position has been led chiefly by prevailing medical data. In Singapore, however, there has been an increase in its utilization supported primarily by social, religious and cultural factors expressly to prolong life of the terminally ill patient. Here this article will seek to review the ethical and clinical impact of this treatment and provide some understanding for such decisions in the light of the Duty of Palliative Care [DoPC]. Complemented by virtue ethics theory, the DoPC highlights and seeks to realize the individual case specific goals of care that maximize comfort and quality of life of the patient in the face of rapid attenuation of treatment options and the eminence of the final outcome by considering each of these factors individually in order to provide the best outcome for the patient and the family. PMID- 21642336 TI - Effects of methotrexate on the expression of the translational isoforms of glucocorticoid receptors alpha and beta: correlation with methotrexate efficacy in rheumatoid arthritis patients. AB - OBJECTIVES: To test the effect of MTX on the expression of glucocorticoid receptor (GR) alpha and beta isoforms AB, C and D in peripheral blood mononuclear cells (PBMCs) in culture, from newly diagnosed RA patients and to evaluate whether the test results correlate with patients' subsequent response to MTX treatment. METHODS: Twenty patients with early active RA were enrolled. Patients who had previously received any DMARD or cytotoxic agent, or who had received CSs in the 6 months before enrolment were excluded. PBMCs from all patients were obtained and cultured in the presence and absence of MTX (10(-4), 10(-6) and 10( 8) M). The expression of GR isoforms was evaluated by western blot. After blood samples were taken, patients entered a 24-week study receiving MTX, diclofenac and prednisone (10 mg/day). At Week 24, the ACR core set of disease activity measures was calculated and a correlation between the MTX effect on patients' PBMC GR expression in vitro and the ACR response was evaluated. RESULTS: MTX 10( 6) M in the culture medium induced the expression of the PBMC isoform AB of GRalpha (P = 0.009). Other GR isoforms were unaffected. The magnitude of the induced expression correlated with the ACR response to treatment at Week 24 of therapy (r = 0.92, P = 0.00003). CONCLUSION: MTX in vitro induces greater expression of GRalphaAB isoform in PBMC from RA patients who later respond to MTX treatment than in non-responding patients. This may have clinical applications for predicting MTX efficacy in RA patients. PMID- 21642337 TI - Compromised weight gain, milk intake, and feeding behavior in breastfed newborns of depressive mothers. AB - OBJECTIVE: To explore depressed mood in the breastfeeding dyad. METHOD: N = 50 mothers of 12-day-olds reported depressed mood (EPDS) and anxiety (STAI), then were videotaped while breastfeeding. Infants were weighed before and after breastfeeding. RESULTS: An ANCOVA on weight gain, which controlled for infant age and birth weight, found EPDS inversely related to weight gain. Following a significant MANCOVA on infant biobehavioral measures, ANCOVAs which controlled for birth weight, age, hunger at time of testing, and degree of exclusivity in breastfeeding, found EPDS inversely related to infants' milk intake and latch quality. Following a significant MANOVA on maternal behaviors, ANOVAs revealed EPDS inversely related to mothers' sensitive positioning and touch frequency. Contrastingly, anxiety was associated with increased touch. CONCLUSION: In the breastfed newborn, mothers' depressed mood is associated with lesser weight gain, lower milk intake, poorer latch to the breast, and receiving less frequent touch and less-sensitive positioning at the breast by mother. PMID- 21642338 TI - Savoryellales (Hypocreomycetidae, Sordariomycetes): a novel lineage of aquatic ascomycetes inferred from multiple-gene phylogenies of the genera Ascotaiwania, Ascothailandia, and Savoryella. AB - The taxonomic placement of freshwater and marine Savoryella species has been widely debated, and the genus has been tentatively assigned to various orders in the Sordariomycetes. The genus is characterized as possessing paraphyses that deliquesce early, elongate, clavate to cylindrical asci with a poorly developed apical ring and versicolored, three-septate ascospores. We performed two combined phylogenetic analyses of different genes: (i) partial small subunit rRNA (SSU), large subunit rRNA (LSU), DNA-dependent RNA polymerase II largest subunit (rpb2) dataset and (ii) SSU rDNA, LSU rDNA, DNA-dependent RNA polymerase II largest subunit (rpb1 and rpb2), translation elongation factor 1-alpha (tef1), the 5.8S ribosomal DNA (5.8S rDNA) dataset. Our results indicate that Savoryella species formed a monophyletic group within the Sordariomycetes but showed no affinity to the Hypocreales, Halosphaeriales (now Microascales), Sordariales and Xylariales, despite earlier assignments to these orders. Savoryella, Ascotaiwania and Ascothailandia (and its anamorph, Canalisporium) formed a new lineage that has invaded both marine and freshwater habitats, indicating that these genera share a common ancestor and are closely related. Because they show no clear relationship with any named order we erect a new order Savoryellales in the subclass Hypocreomycetidae, Sordariomycetes. The genera Savoryella and Ascothailandia are monophyletic, while the position of Ascotaiwania is unresolved. All three genera are phylogenetically related and form a distinct clade similar to the unclassified group of marine ascomycetes comprising the genera Swampomyces, Torpedospora and Juncigera (TBM clade: Torpedospora/Bertia/Melanospora) in the Hypocreomycetidae incertae sedis. PMID- 21642339 TI - Development of molecular markers and preliminary investigation of the population structure and mating system in one lineage of black morel (Morchella elata) in the Pacific Northwestern USA. AB - Phylogenetic analysis of LSU/ITS sequence data revealed two distinct lineages among 44 morphologically similar fruiting bodies of natural black morels (Morchella elata group) sampled at three non-burn locations in the St Joe and Kanisku National Forests in northern Idaho. Most of the sampled isolates (n = 34) represented a dominant LSU/ITS haplotype present at all three sites and identical to the Mel-12 phylogenetic lineage (GU551425) identified in a previous study. Variation at 1-3 nucleotide sites was detected among a small number of isolates (n = 6) within this well supported clade (94%). Four isolates sampled from a single location were in a well supported clade (97%) distinct from the dominant haplotypes and may represent a previously un-sampled, cryptic phylogenetic species. Species-specific SNP and SCAR markers were developed for Mel-12 lineage isolates by cloning and sequencing AFLP amplicons, and segregation of AFLP markers were studied from single ascospore isolates from individual fruiting bodies. Based on the segregation of AFLP markers within single fruiting bodies, split decomposition analyses of two SCAR markers, and population genetic analyses of SNP, SCAR, and AFLP markers, it appears that members of the Morchella sp. Mel 12 phylogenetic lineage are heterothallic and outcross in nature similar to yellow morels. This is the first set of locus-specific molecular markers that has been developed for any Morchella species, to our knowledge. These markers will prove to be valuable tools to study mating system, gene flow and genetic structure of black morels at various spatial scales with field-collected fruiting bodies and eliminate the need to culture samples in vitro. PMID- 21642340 TI - Molecular identification of Pilobolus species from Yellowstone National Park. AB - Pilobolus, a widely distributed coprophilous fungus, grows on herbivore dung. Species of Pilobolus traditionally are described with imprecise morphological characteristics potentially leading to misidentification. In this study we used PCR analysis of taxonomically informative sequences to provide more consistent species identification from isolates obtained in Yellowstone National Park. We collected Pilobolus park-wide from six taxa of herbivores over 9 y. Multiple transfers of single sporangium isolates provided pure cultures from which DNA was extracted. Sequence analysis of internal transcribed spacer (ITS) regions of DNA that code for rRNA genes were used to distinguish among similar species. We describe several species of Pilobolus associated with herbivores in various habitats, including two species not previously reported, P. heterosporus and P. sphaerosporus. Our results show that phylogenetic species identification of Pilobolus based on sequence analysis of pure culture isolates provides a more reliable means of identifying species than traditional methods. PMID- 21642341 TI - A new lineage in Pucciniomycotina: class Tritirachiomycetes, order Tritirachiales, family Tritirachiaceae. AB - Based on multiple gene analyses (nuclear large subunit, nuclear small subunit, internal transcribed spacer region including the 5.8 s subunit rDNA, and translation elongation factor 1alpha) and septal pore ultrastructure we describe a new lineage of Pucciniomycotina consisting of a new class, Tritirachiomycetes, new order, Tritirachiales, and new family, Tritirachiaceae. Tritirachium dependens, T. oryzae, T. roseum (reintroduced), T. cinnamomeum and two unidentified species are recognized. Phylogenetic analyses do not support existing morphological circumscription of some species, and the available evidence suggests that morphological evaluation alone is not adequate for species identification. PMID- 21642342 TI - Genetic diversity of Rhizoctonia solani associated with potato tubers in France. AB - The soilborne fungus Rhizoctonia solani is a pathogen of many plants and causes severe damage in crops around the world. Strains of R. solani from the anastomosis group (AG) 3 attack potatoes, leading to great yield losses and to the downgrading of production. The study of the genetic diversity of the strains of R. solani in France allows the structure of the populations to be determined and adapted control strategies against this pathogen to be established. The diversity of 73 French strains isolated from tubers grown in the main potato seed production areas and 31 strains isolated in nine other countries was assessed by phylogenetic analyses of (i) the internal transcribed spacer sequences (ITS1 and ITS2) of ribosomal RNA (rRNA), (ii) a part of the gene tef-1alpha and (iii) the total DNA fingerprints of each strain established by amplified fragment length polymorphism (AFLP). The determination of the AGs of R. solani based on the sequencing of the ITS region showed three different AGs among our collection (60 AG 3 PT, 8 AG 2-1 and 5 AG 5). Grouping of the strains belonging to the same AG was confirmed by sequencing of the gene tef-1alpha used for the first time to study the genetic diversity of R. solani. About 42% of ITS sequences and 72% of tef-1alpha sequences contained polymorphic sites, suggesting that the cells of R. solani strains contain several copies of ITS and the tef-1alpha gene within the same nucleus or between different nuclei. Phylogenetic trees showed a greater genetic diversity within AGs in tef-1alpha sequences than in ITS sequences. The AFLP analyses showed an even greater diversity among the strains demonstrating that the French strains of R. solani isolated from potatoes were not a clonal population. Moreover there was no relationship between the geographical origins of the strains or the variety from which they were isolated and their genetic diversity. PMID- 21642343 TI - A taxonomic revision of the North American species of Lepraria s.l. that produce divaricatic acid, with notes on the type species of the genus L. incana. AB - The divaricatic acid-producing populations of Lepraria in North America north of Mexico are revised with traditional morphological characters, chemistry, ecology, biogeography, and ITS1, 5.8S, and ITS2 sequence data. Three taxa are accepted: L. cryophila, L. hodkinsoniana sp. nov. and L. pacifica sp. nov. Both Lepraria crassissima and L. incana are excluded from the study area. Noncryptic, semicryptic and fully cryptic species concepts in Lepraria are discussed with emphasis on the practical integration of molecular characters into taxonomic frameworks based on non-molecular characters. PMID- 21642344 TI - Phylogenetic diversity of fungal communities in areas accessible and not accessible to tourists in Naracoorte Caves. AB - The fungal diversity in areas accessible and not accessible to tourists at UNESCO World Heritage-listed Naracoorte Caves was investigated with culture-dependent and culture-independent techniques for assistance in cave management protocol development. The caves were selected based on tourist numbers and configurations: Stick Tomato (open, high numbers), Alexandra (lockable openings, high numbers) and Strawhaven (control; no access). Culture-based survey revealed Ascomycota dominance irrespective of sampling area with Microascales (Trichurus sp.) being most frequently isolated. Some Hypocreales-like sequences belonging to Fusarium sp., Trichoderma sp. and Neonectria sp. (Stick Tomato) were cultured only from areas not accessible to tourists. These orders also were detected by DGGE assay irrespective of sampling area. The predominance of Ascomycota (especially Microascales) suggested their important ecological roles in these caves. Culture independent analysis showed higher Shannon fungal diversity values (from ITS based DGGE profiles) in tourist-accessible areas of these caves than in inaccessible areas with the fungal community banding patterns being substantially different in Stick Tomato Cave. Further investigations are needed to determine the cause of the differences in the fungal communities of Stick Tomato Cave, although cave-related factors such as use, configuration and sediment heterogeneity might have contributed to these differences. PMID- 21642345 TI - Identification of new polymorphic microsatellite markers in the NA1 and NA2 lineages of Phytophthora ramorum. AB - Phytophthora ramorum is a recently introduced pathogen in Europe and North America consisting of three clonal lineages. Due to the limited intralineage genetic variation, only a few polymorphic markers are available for use in studies involving the epidemiology and evolution of P. ramorum. A total of 159 primer pairs for candidate polymorphic SSR loci were tested with universal labeling. Four polymorphic microsatellite loci were identified within the NA1 lineage and one within the NA2 lineage, demonstrating the power and flexibility of the screening technique. The markers may significantly increase the number of genotypes that can be identified and as such can help better characterize the North American lineages of P. ramorum. PMID- 21642346 TI - Effect of light on in vivo urediniospore germination, lesion development and sporulation of Puccinia hemerocallidis on daylily and Puccinia pelargoniizonalis on geranium. AB - The presence of rusts of daylily and geranium caused respectively by Puccinia hemerocallidis and P. pelargoniizonalis can result in reduced value of these ornamental crops. Experiments were conducted to determine the effects of fluorescent light and sunlight on urediniospore germination, germ tube elongation, lesion development and sporulation of the two fungal pathogens on detached leaves and whole plants. Exposure of dry or hydrated urediniospores of P. hemerocallidis to cool white fluorescent light (600 MUmol s(-1) m(-2)) or to sunlight (950-1910 MUmol s(-1) m(-2)) for 2 h or 4 h significantly reduced germination and germ tube elongation on detached daylily leaves. Germination but not germ tube elongation of hydrated urediniospores of P. pelargoniizonalis on detached geranium leaves was significantly reduced when exposed to fluorescent light for 2 h or 4 h. A 4 h exposure to either light source significantly reduced lesion development of P. hemerocallidis on detached daylily leaves with fewer lesions developing from hydrated compared to dry urediniospores. Sunlight exposures of 1 h and 2 h of hydrated and dry urediniospores respectively significantly reduced lesion development by either fungus on whole plants. Increasing exposure to fluorescent light negatively affected sporulation of P. hemerocallidis and P. pelargoniizonalis. Complete suppression of sporulation was not observed for either fungus with up to a 24 h exposure to fluorescent light. Light exposure negatively affected disease development by P. hemerocallidis and P. pelargoniizonalis. Exposure to high light intensities may affect spread of rust diseases on ornamental plants. PMID- 21642347 TI - A six-gene phylogeny reveals the evolution of mode of infection in the rice blast fungus and allied species. AB - The family Magnaporthaceae contains devastating fungal cereal and grass pathogens, such as Magnaporthe oryzae (rice blast fungus, formerly known as M. grisea), M. poae (summer patch pathogen of turf grasses) and Gaeumannomyces graminis (take-all fungus of various cereals and grasses), which are popular model organisms in fungal biology and host-pathogen interaction studies. Despite their ecological and economic importance, the phylogenetic relationships among the constituent species remain ambiguous due to the lack of convincing morphological characters and paucity of molecular data for the majority of the non-model species in the family. In this study our multilocus phylogeny suggests that both Magnaporthe and Gaeumannomyces are polyphyletic genera. The phylogeny also provides insights into fungal biology and pathogenesis. Magnaporthe oryzae formed a basal clade, while M. poae and M. rhizophila formed another well supported clade with G. incrustans and G. graminis. The basal species infect both root and aerial parts of the plant host, while the aerial infection capacity seems to be lost in the taxa of the latter clade. The phylogeny is corroborated by evolution of the anamorphs and a cAMP-dependent protein kinase (CPKA) gene. Magnaporthe oryzae produces Pyricularia, while taxa in the latter clade all produce Phialophora-like anamorphs. CPKA is present in animals and many fungal lineages with various functions. In M. oryzae CPKA is essential for the formation of functional appressoria for leaf penetration. In root-infecting G. graminis var. tritici and M. poae however only non-functional CPKA homologous pseudogenes were found in their genomes. The study indicates that anamorphic and ecological features are more informative than the teleomorphic characters in defining monophyletic groups among these taxa. PMID- 21642348 TI - Phylogenetic affiliations of members of the heterogeneous lichen-forming fungi of the genus Lecidea sensu Zahlbruckner (Lecanoromycetes, Ascomycota). AB - The genus Lecidea Ach. sensu lato (sensu Zahlbruckner) includes almost 1200 species, out of which only 100 species represent Lecidea sensu stricto (sensu Hertel). The systematic position of the remaining species is mostly unsettled but anticipated to represent several unrelated lineages within Lecanoromycetes. This study attempts to elucidate the phylogenetic placement of members of this heterogeneous group of lichen-forming fungi and to improve the classification and phylogeny of Lecanoromycetes. Twenty-five taxa of Lecidea sensu lato and 22 putatively allied species were studied in a broad selection of 268 taxa, representing 48 families of Lecanoromycetes. Six loci, including four ribosomal and two protein-coding genes for 315- and 209-OTU datasets were subjected to maximum likelihood and Bayesian analyses. The resulting well supported phylogenetic relationships within Lecanoromycetes are in agreement with published phylogenies, but the addition of new taxa revealed putative rearrangements of several families (e.g. Catillariaceae, Lecanoraceae, Lecideaceae, Megalariaceae, Pilocarpaceae and Ramalinaceae). As expected, species of Lecidea sensu lato and putatively related taxa are scattered within Lecanoromycetidae and beyond, with several species nested in Lecanoraceae and Pilocarpaceae and others placed outside currently recognized families in Lecanorales and orders in Lecanoromycetidae. The phylogenetic affiliations of Schaereria and Strangospora are outside Lecanoromycetidae, probably with Ostropomycetidae. All species referred to as Lecidea sensu stricto based on morphology (including the type species, Lecidea fuscoatra [L.] Ach.) form, with Porpidia species, a monophyletic group with high posterior probability outside Lecanorales, Peltigerales and Teloschistales, in Lecanoromycetidae, supporting the recognition of order Lecideales Vain. in this subclass. The genus name Lecidea must be redefined to apply only to Lecidea sensu stricto and to include at least some members of the genus Porpidia. Based on morphological and chemical similarities, as well as the phylogenetic relationship of Lecidea pullata sister to Frutidella caesioatra, the new combination Frutidella pullata is proposed here. PMID- 21642349 TI - Commentary: Money and models: double-edged swords. PMID- 21642350 TI - Cohort Profile: the Corogene study. PMID- 21642351 TI - Automatic response activation in sequential affective priming: an ERP study. AB - Affective priming effects denote faster responses when two successively presented affective stimuli match in valence than when they mismatch. Two mechanisms have been proposed for their explanation: (i) Priming of affective information within a semantic network or distributed memory system (semantic priming). (ii) Automatic activation of the evaluative response through the affective prime (response priming). In this experiment, we sought more direct evidence for prime induced response activations with measurement of the lateralized readiness potential (LRP). Onset of the stimulus-locked LRP was earlier in affectively congruent trials than in incongruent trials. In addition, priming modulated the LRP-amplitude of slow responses, indicating greater activation of the incorrect response hand in affectively incongruent trials. Onset of the response-locked LRP and peak latency of the P300 component were not modulated by priming but the amplitude of the N400 component was. In combination, these results suggest that both, semantic priming and response priming constitute affective priming effects in the evaluative categorization task. PMID- 21642352 TI - The importance of actions and the worth of an object: dissociable neural systems representing core value and economic value. AB - Neuroeconomic research has delineated neural regions involved in the computation of value, referring to a currency for concrete choices and decisions ('economic value'). Research in psychology and sociology, on the other hand, uses the term 'value' to describe motivational constructs that guide choices and behaviors across situations ('core value'). As a first step towards an integration of these literatures, we compared the neural regions computing economic value and core value. Replicating previous work, economic value computations activated a network centered on medial orbitofrontal cortex. Core value computations activated medial prefrontal cortex, a region involved in the processing of self-relevant information and dorsal striatum, involved in action selection. Core value ratings correlated with activity in precuneus and anterior prefrontal cortex, potentially reflecting the degree to which a core value is perceived as internalized part of one's self-concept. Distributed activation pattern in insula and ACC allowed differentiating individual core value types. These patterns may represent evaluation profiles reflecting prototypical fundamental concerns expressed in the core value types. Our findings suggest mechanisms by which core values, as motivationally important long-term goals anchored in the self-schema, may have the behavioral power to drive decisions and behaviors in the absence of immediately rewarding behavioral options. PMID- 21642353 TI - Response inhibition results in the emotional devaluation of faces: neural correlates as revealed by fMRI. AB - Although it is well established that prior experience with faces determines their subsequent social-emotional evaluation, recent work shows that top-down inhibitory mechanisms, including response inhibition, can lead to social devaluation after even a single, brief exposure. These rapidly induced effects indicate interplay among perceptual, attentional, response-selection and social emotional networks; yet, the brain mechanisms underlying this are not well understood. This study used functional magnetic resonance imaging (fMRI) to investigate the neural mechanism mediating the relationship between inhibitory control and emotional devaluation. Participants performed two tasks: (i) a Go/No Go task in response to faces and (ii) a trustworthiness rating task involving the previously seen faces. No-Go faces were rated as significantly less trustworthy than Go faces. By examining brain activations during Task 1, behavioral measures and brain activations obtained in Task 2 could be predicted. Specifically, activity in brain areas during Task 1 associated with (i) executive control and response suppression (i.e. lateral prefrontal cortex) and (ii) affective responses and value representation (i.e. orbitofrontal cortex), systematically covaried with behavioral ratings and amygdala activity obtained during Task 2. The present findings offer insights into the neural mechanisms linking inhibitory processes to affective responses. PMID- 21642354 TI - p54nrb is a new regulator of progression of malignant melanoma. AB - Nuclear RNA-binding protein p54(nrb) and its murine homolog NonO are known to be involved in a variety of nuclear processes including transcription and RNA processing. Melanoma inhibitory activity (MIA) has been shown to play an essential role in the progression of malignant melanoma and to influence melanoma associated molecules and pathways in the early tumor formation steps. Interestingly, recent studies suggest that MIA is a regulator of p54(nrb). Here, we show that p54(nrb) is strongly expressed and localized in the nucleus of both melanoma cell lines and melanoma tissue samples compared with normal human melanocytes or normal skin, respectively. Furthermore, all tested melanoma cell lines revealed strong p54(nrb) promoter activity. Treatment with MIA-specific small interfering RNAs showed an influence of MIA on p54(nrb) expression on both messenger RNA (mRNA) and protein level. Knockdown of p54(nrb) protein in melanoma cell lines led to reduced proliferation rates and to a strong decrease in their migratory potential. In addition, attachment to laminin and poly-l-lysine was significantly increased. We could identify Connexin-43 (Cx-43) as a downstream target molecule of p54(nrb) as knockdown of p54(nrb) resulted in enhanced Cx-43 mRNA and protein levels. As a confirmation of these findings, melanoma cell lines showed very low Cx-43 expression levels compared with melanocytes. Our results demonstrate that p54(nrb) is highly expressed in malignant melanoma and, as a MIA target molecule, it seems to be involved in the development and progression of malignant melanoma. PMID- 21642355 TI - Inhibition of aldose reductase prevents colon cancer metastasis. AB - Colon cancer is the third most common cause of cancer and is the second leading cause of cancer deaths in the USA. Although inhibition of aldose reductase (AR) is known to prevent human colon cancer cell growth in nude mice xenografts, the role of AR in the regulation of cancer metastasis is not known. We now demonstrate the mechanisms by which AR regulates colon cancer metastasis in vitro and in vivo. Inhibition of AR prevented the epidermal growth factor (EGF) or fibroblast growth factor (FGF)-induced migration and invasion of human colon cancer (HT29; KM20) cells by >70% and also inhibited (>80%) the adhesion of the cancer cells to endothelial cells. Treatment of endothelial cells with AR inhibitors significantly (~85%) downregulated the EGF or FGF-induced expression of Inter-Cellular Adhesion Molecule-1, Vascular cell adhesion molecule-1 and vascular endothelial-cadherin. Furthermore, liver metastasis of green fluorescent protein-labeled KM20 cells injected into the spleen of athymic nude mice was significantly (>65%) prevented by AR inhibitor, fidarestat or ARsiRNA delivered systemically into the mice. Similar results were observed with HT29 cells. AR inhibition or ablation also prevented (70-90%) the increase in the levels of matrix metalloproteinase-2, cyclin D1, CD31, CD34 and the activation of nuclear factor-kappa-binding protein in metastatic liver. Thus, our results indicate that AR regulates cancer cell adhesion, invasion and migration events which initiate metastasis and therefore, AR inhibition could be a novel therapeutic approach for the prevention of colon cancer metastasis. PMID- 21642356 TI - Fyn requires HnRNPA2B1 and Sam68 to synergistically regulate apoptosis in pancreatic cancer. AB - PURPOSE: The Src family kinase Fyn, heterogenous nuclear ribonucleoprotein (HnRNP) A2B1 and Sam68 are thought to be associated with the metastasis of tumors, but their roles in the regulation of apoptosis remain unclear. This study investigated the role of Fyn and its potential relationship with HnRNPA2B1 and Sam68 in the regulation of apoptosis in pancreatic cancer. Experimental design. We examined both the activity of Fyn and the expression of HnRNPA2B1 in human pancreatic cancer tissues and systematically investigated the apoptotic mechanisms induced by Fyn activity using multiple experimental approaches. RESULTS: We found that Fyn activity was increased in metastatic pancreatic cancer tissues. In the pancreatic cancer BxPc3 cell line, the inhibition of Fyn activity by kinase-dead Fyn downregulated HnRNPA2B1 expression. Further analysis showed that HnRNPA2B1 expression was associated with pancreatic cancer progression. In BxPc3 cells, HnRNPA2B1 bound to Bcl-x messenger RNA (mRNA), which affected splicing and therefore, the formation of Bcl-x(s). Downregulation of HnRNPA2B1 by RNA interference (RNAi) resulted in the increased formation of the pro-apoptotic Bcl-x(s) and promoted apoptosis of BxPc3 cells. In addition, deactivation of Fyn in BxPc3 cells reduced Sam68 phosphorylation. This resulted in increased binding between Sam68 and Bcl-x mRNA, promoting the formation of the anti-apoptotic Bcl x(L). The knockdown of Sam68 by RNAi also increased the formation of Bcl-x(L). Finally, HnRNPA2B1 overexpression or Sam68 knockdown could rescue pancreatic cancer cells from apoptosis. CONCLUSION: Our results suggest a mechanism by which Fyn requires HnRNPA2B1 and Sam68 to coordinate and regulate apoptosis, thus promoting the proliferation and metastasis of pancreatic cancer. PMID- 21642357 TI - Hypoxia-increased RAGE and P2X7R expression regulates tumor cell invasion through phosphorylation of Erk1/2 and Akt and nuclear translocation of NF-{kappa}B. AB - The role of hypoxia in regulating tumor progression is still controversial. Here, we demonstrate that, similarly to what previously observed by us in human prostate and breast tumor samples, hypoxia increases expression of the receptor for advanced glycation end products (RAGE) and the purinergic receptor P2X7 (P2X7R). The role of hypoxia was shown by the fact that hypoxia-inducible factor (HIF)-1alpha silencing downregulated RAGE and P2X7R protein levels as well as nuclear factor-kappaB (NF-kappaB) expression. In contrast, NF-kappaB silencing reduced P2X7R expression without affecting RAGE protein levels or nuclear accumulation of HIF-1alpha. Treatment of hypoxic tumor cells with HMGB1 and BzATP ligands, respectively, of RAGE and P2X7R, activated a signaling pathway that, through Akt and Erk phosphorylation, determines nuclear accumulation of NF-kappaB and increases cell invasion. Inhibition of Akt by SH5 and Erk by INH1 prevented both nuclear translocation of NF-kappaB and cell invasion. Moreover, silencing RAGE and P2X7R abolished nuclear accumulation of NF-kappaB as well as cell invasion without affecting HIF-1alpha stabilization. Once in the nucleus, NF kappaB would contribute to cell survival and invasion under hypoxia, by maintaining RAGE and P2X7R expression levels and matrix metalloproteinases 2 and 9 synthesis. These results show that, hypoxia can upregulate expression levels of membrane receptors that, by binding extracellular molecules eventually released by necrotic cells, contribute to the increased invasiveness of transformed tumor cells. Moreover, these observations strengthen our working hypothesis that upregulation of damage-associated molecular patterns receptors by HIF-1alpha represents the crucial event bridging hypoxia and inflammation in obtaining the malignant phenotype. PMID- 21642358 TI - Common genetic variants related to genomic integrity and risk of papillary thyroid cancer. AB - DNA damage is an important mechanism in carcinogenesis, so genes related to maintaining genomic integrity may influence papillary thyroid cancer (PTC) risk. Candidate gene studies targeting some of these genes have identified only a few polymorphisms associated with risk of PTC. Here, we expanded the scope of previous candidate studies by increasing the number and coverage of genes related to maintenance of genomic integrity. We evaluated 5077 tag single-nucleotide polymorphisms (SNPs) from 340 candidate gene regions hypothesized to be involved in DNA repair, epigenetics, tumor suppression, apoptosis, telomere function and cell cycle control and signaling pathways in a case-control study of 344 PTC cases and 452 matched controls. We estimated odds ratios for associations of single SNPs with PTC risk and combined P values for SNPs in the same gene region or pathway to obtain gene region-specific or pathway-specific P values using adaptive rank-truncated product methods. Nine SNPs had P values <0.0005, three of which were in HDAC4 and were inversely related to PTC risk. After multiple comparisons adjustment, no SNPs remained associated with PTC risk. Seven gene regions were associated with PTC risk at P < 0.01, including HUS1, ALKBH3, HDAC4, BAK1, FAF1_CDKN2C, DACT3 and FZD6. Our results suggest a possible role of genes involved in maintenance of genomic integrity in relation to risk of PTC. PMID- 21642359 TI - Heterogenous effect of androgen receptor CAG tract length on testicular germ cell tumor risk: shorter repeats associated with seminoma but not other histologic types. AB - Increasing rates of testicular germ cells tumors (TGCTs) overtime suggest that environmental factors are involved in disease etiology, but familial risk and genome-wide association studies implicate genetic factors as well. We investigated whether variation in the functional CAG(n) polymorphism in the androgen receptor (AR) gene is associated with TGCT risk, using data from a population-based family study. We estimated odds ratios (OR) and 95% confidence intervals (CI) for the association of CAG repeat length and TGCT risk using matched pairs logistic regression. Analyses of 273 TGCT case-mother pairs revealed no association between AR CAG repeat length and overall TGCT risk. However, risk of seminoma was significantly associated with shorter CAG repeat length [CAG 20-21 versus CAG <= 19: OR = 0.82 (95% CI: 0.43-1.58), CAG 22-23 versus CAG <= 19: OR = 0.39 (95% CI: 0.19-0.83) and CAG >= 24 versus CAG <= 19: OR = 0.42 (95% CI: 0.20-0.86)], with a highly significant trend over these four categories of decreasing CAG repeat length (P(trend) = 0.0030). This is the first report of a statistically significant association between AR CAG repeat length and seminoma risk, suggesting that increased AR transactivation may be involved in development of seminoma and/or progression of carcinoma in situ/intratubular germ cell neoplasia unclassified to seminoma. This result provides a rationale whereby androgenic environmental compounds could contribute to increases in TGCT incidence, and identifies for the first time a potential biological pathway influencing whether TGCTs achieve seminomatous versus nonseminomatous histology, a clinically and biologically important distinction. PMID- 21642360 TI - Social networks--are they good for your health? The era of Facebook and Twitter. PMID- 21642361 TI - More fundamental issues in diabetic ketoacidosis. PMID- 21642362 TI - Sub-hepatic cholesterol stones indicate a sinister underlying diagnosis. PMID- 21642363 TI - Asymmetric dimethylarginine and clinical outcomes in chronic kidney disease. AB - BACKGROUND AND OBJECTIVES: Elevated plasma level of asymmetric dimethylarginine (ADMA) have been reported to be associated with endothelial dysfunction and atherosclerosis risk factors, and may predict cardiovascular events in patients with ESRD. In this study, we aimed to assess the association between plasma ADMA and long-term outcome in a cohort of patients with stage 3 to 4 chronic kidney disease (CKD). DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: From July 2006 to June 2009, 298 consecutive patients with stage 3 to 4 CKD scheduled to undergo coronary angiography were recruited. Plasma ADMA levels were determined using HPLC. RESULTS: The mean age was 73 +/- 10 years. Approximately half of the patients had diabetes and 88 patients had proteinuria. The baseline estimated GFR (eGFR) was 44 +/- 13 ml/min per 1.73 m2. The plasma ADMA levels of the patients with proteinuria were significantly higher than those without. The plasma ADMA levels correlated significantly with eGFR. During the median follow-up period of 2.7 years, we observed 26 all-cause deaths, 12 nonfatal myocardial infarctions, and 2 strokes. Multivariate Cox analysis revealed that an increase of 0.1 MUmol/L in plasma ADMA level was associated with a 37% increased risk of the composite outcomes of all-cause deaths, nonfatal myocardial infarctions, and strokes. CONCLUSIONS: In this elder and high-risk population with stage 3 to 4 CKD, high plasma ADMA level was associated with low eGFR and macroalbuminuria. Furthermore, high plasma ADMA level appeared to be an independent predictor of long-term outcome. PMID- 21642364 TI - Inflammation and asymmetric dimethylarginine for predicting death and cardiovascular events in ESRD patients. AB - BACKGROUND: Endothelial dysfunction as assessed by asymmetric dimethylarginine (ADMA) and inflammation has been consistently linked to atherosclerosis, death, and cardiovascular (CV) events in ESRD patients. Inflammation amplifies the effect of ADMA on the severity of atherosclerosis in ESRD patients, but it is still unknown whether inflammation and ADMA interact in the high risk of death and CV events in this population. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: In a cohort of 225 hemodialysis patients, we investigated the interaction between inflammatory biomarkers (C-reactive protein and IL-6) and ADMA as predictors of death and CV events over an extended follow-up (13 years). RESULTS: During follow up, 160 patients died, and 123 had CV events. With crude and multiple Cox regression analyses, an interaction was found between inflammation biomarkers and ADMA for explaining death and CV events in ESRD patients. The adjusted hazard ratios (HRs) for death (HR, 2.18; 95% confidence interval [CI], 1.34 to 3.54) and CV outcomes (HR, 2.59; 95% CI, 1.47 to 4.55) of patients with C-reactive protein and ADMA above the median were higher than expected in the absence of interaction under the additive model (1.15 and 1.97, respectively) and significantly higher than in patients with only one biomarker above the median. Data analyses carried out by stratifying patients according to IL-6 provided similar results. CONCLUSIONS: These data support the hypothesis that inflammation amplifies the risk of death and CV events associated with high ADMA levels in ESRD. These analyses further emphasize the need for intervention studies to attenuate inflammation and high ADMA levels in this population. PMID- 21642365 TI - Arteriovenous graft infection: a comparison of thigh and upper extremity grafts. AB - BACKGROUND AND OBJECTIVES: There are a limited number of publications on the features of arteriovenous graft infection in hemodialysis patients. The authors compared the clinical presentation, complications, and outcomes of infections of thigh and upper extremity grafts. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: The authors queried a prospective access database at a large university medical center and identified 132 patients with graft infections (40 in the thigh and 92 in the upper extremity) requiring surgical excision. The authors collected information regarding the microbiology, complications, and clinical outcomes. RESULTS: The two graft groups were similar in age, gender, race, and frequency of diabetes. The median age of infected grafts was 162 days for thigh grafts versus 168 days for upper extremity grafts (P = 0.35). Thigh graft infections were more likely than upper extremity graft infections to be caused by a Gram-negative rod (31% versus 4%; P = 0.003), and more likely to result in a metastatic infection (15% versus 3%; P = 0.02). The duration of hospitalization associated with graft infection was similar (10.8 +/- 5.4 versus 8.7 +/- 6.3 days; P = 0.09). Finally, median catheter dependence was longer after thigh graft than upper arm graft infections (319 versus 237 days; P = 0.04). CONCLUSIONS: As compared with upper extremity graft infections, thigh graft infections requiring excision are more likely to be caused by Gram-negative bacteria and to result in serious metastatic complications. These differences may require different empiric antibiotics and a higher index of suspicion for infection in hemodialysis patients with thigh grafts. PMID- 21642366 TI - Plant surfaces with cuticular folds are slippery for beetles. AB - Plant surfaces covered with three-dimensional (3D) waxes are known to strongly reduce insect adhesion, leading to slippery surfaces. Besides 3D epicuticular waxes, cuticular folds are a common microstructure found on plant surfaces, which have not been quantitatively investigated with regard to their influence on insect adhesion. We performed traction experiments with Colorado potato beetles on five plant surfaces with cuticular folds of different magnitude. For comparison, we also tested (i) smooth plant surfaces and (ii) plant surfaces possessing 3D epicuticular waxes. Traction forces on surfaces with medium cuticular folds, of about 0.5 um in both height and thickness and a spacing of 0.5-1.5 um, were reduced by an average of 88 per cent in comparison to smooth plant surfaces. Traction forces were reduced by the same order of magnitude as on plant surfaces covered with 3D epicuticular waxes. For surface characterization, we performed static contact angle measurements, which proved a strong effect of cuticular folds also on surface wettability. Surfaces possessing cuticular folds of greater magnitude showed higher contact angles up to superhydrophobicity. We hypothesize that cuticular folds reduce insect adhesion mainly due to a critical roughness, reducing the real contact area between the surface and the insect's adhesive devices. PMID- 21642367 TI - Three-dimensional reconstruction of the fast-start swimming kinematics of densely schooling fish. AB - Information transmission via non-verbal cues such as a fright response can be quantified in a fish school by reconstructing individual fish motion in three dimensions. In this paper, we describe an automated tracking framework to reconstruct the full-body trajectories of densely schooling fish using two dimensional silhouettes in multiple cameras. We model the shape of each fish as a series of elliptical cross sections along a flexible midline. We estimate the size of each ellipse using an iterated extended Kalman filter. The shape model is used in a model-based tracking framework in which simulated annealing is applied at each step to estimate the midline. Results are presented for eight fish with occlusions. The tracking system is currently being used to investigate fast-start behaviour of schooling fish in response to looming stimuli. PMID- 21642368 TI - Stem cells and the endocrine pancreas. AB - BACKGROUND: Diabetes can be treated by beta-cell replacement therapy but the supply of graft material from human donors is too limited to make a significant clinical impact. Substitute beta-cells generated from stem cell populations offer a potential source for the large numbers of cells required. SOURCES OF DATA: Primary peer-reviewed reports of experimental studies. AREAS OF AGREEMENT: Embryonic stem cells and/or induced pluripotent stem (iPS) cells are currently the most promising starting populations from which to generate large numbers of beta-cells. Differentiation protocols that recapitulate in vivo development generate insulin-expressing cells in vitro. AREAS OF CONTROVERSY: Differentiation outcomes may depend on the source of the initial pluripotent cells. The insulin expressing cells are not fully functional. In vivo maturation is inconsistent and not well understood. AREAS TIMELY FOR DEVELOPING RESEARCH: Improvement of current protocols for complete in vitro differentiation to a functional beta-cell phenotype. Systematic analysis to identify the most appropriate starting material. Improved purification methods to ensure safety of material for clinical transplantation. PMID- 21642369 TI - Management of alert messages in the remote monitoring of implantable cardioverter defibrillators and pacemakers: an Italian single-region study. AB - AIMS: The remote monitoring of pacemakers and implantable cardioverter defibrillators (ICDs) promotes accurate supervision of the patient and device. Alert settings appear to be a crucial parameter of its efficacy. The purpose of our study was to evaluate various settings for alerts and alert message management in patients with pacemakers and ICDs. METHODS AND RESULTS: We prospectively evaluated 282 patients (181 males, 101 females, mean age 72 +/- 11 years) who were referred to nine electrophysiological centres in the Veneto region of Italy for a pacemaker (76 patients) or ICD (206 patients) implant in which remote monitoring was activated (Biotronik Home Monitoring(r), Biotronik, Berlin, Germany). During a mean follow-up of 382 +/- 261 days, we received 106,039 transmissions. In the pacemaker group, the alerts that were most frequently activated were those concerning battery exhaustion, surveying impedances, sensing and threshold measurements, as well as missing transmissions. In the ICD/ICD-cardiac resynchronization therapy group, the alerts nearly always activated were those concerning a detection setoff, battery exhaustion, critical values of impedance, or ineffective maximum energy shock. In both groups, the alarms for heart rate monitoring and supraventricular arrhythmia were activated in fewer cases at higher variability among centres. CONCLUSIONS: Our study demonstrates that more attention is paid to critical technical data than to patients' clinical profiles, probably to limit an excessive flow of data into the centre. Accurate alert settings, personalized to the patients' features, are essential for easier and more effective management of patients who are followed remotely. PMID- 21642370 TI - Comparison of the 2- and 6-minute walk test in multiple sclerosis. AB - The 6-minute walk test (6MWT) is often used to assess walking distance in multiple sclerosis (MS), but can be both time consuming for the investigator and exhausting for people with MS (pwMS). The present report compared the 6MWT scores of 40 ambulatory pwMS with their scores on the shorter 2-minute walk test (2MWT). The 2MWT estimated the 6MWT results with a mean relative error of 5% (R(2) = 0.96; p < 0.01). As the last 4-minute period of the 6MWT seems redundant, the 2MWT may be considered as a practical replacement for the 6MWT in routine clinical assessment. PMID- 21642371 TI - On dimensionality of the DASH. PMID- 21642372 TI - The Wnt inhibitory factor 1 (WIF1) is targeted in glioblastoma and has a tumor suppressing function potentially by induction of senescence. AB - Gene expression-based prediction of genomic copy number aberrations in the chromosomal region 12q13 to 12q15 that is flanked by MDM2 and CDK4 identified Wnt inhibitory factor 1 (WIF1) as a candidate tumor suppressor gene in glioblastoma. WIF1 encodes a secreted Wnt antagonist and was strongly downregulated in most glioblastomas as compared with normal brain, implying deregulation of Wnt signaling, which is associated with cancer. WIF1 silencing was mediated by deletion (7/69, 10%) or epigenetic silencing by promoter hypermethylation (29/110, 26%). Co-amplification of MDM2 and CDK4 that is present in 10% of glioblastomas was associated in most cases with deletion of the whole genomic region enclosed, including the WIF1 locus. This interesting pathogenetic constellation targets the RB and p53 tumor suppressor pathways in tandem, while simultaneously activating oncogenic Wnt signaling. Ectopic expression of WIF1 in glioblastoma cell lines revealed a dose-dependent decrease of Wnt pathway activity. Furthermore, WIF1 expression inhibited cell proliferation in vitro, reduced anchorage-independent growth in soft agar, and completely abolished tumorigenicity in vivo. Interestingly, WIF1 overexpression in glioblastoma cells induced a senescence-like phenotype that was dose dependent. These results provide evidence that WIF1 has tumor suppressing properties. Downregulation of WIF1 in 75% of glioblastomas indicates frequent involvement of aberrant Wnt signaling and, hence, may render glioblastomas sensitive to inhibitors of Wnt signaling, potentially by diverting the tumor cells into a senescence-like state. PMID- 21642373 TI - Two novel determinants of etoposide resistance in small cell lung cancer. AB - Patient survival in small cell lung cancer (SCLC) is limited by acquired chemoresistance. Here we report the use of a biologically relevant model to identify novel candidate genes mediating in vivo acquired resistance to etoposide. Candidate genes derived from a cDNA microarray analysis were cloned and transiently overexpressed to evaluate their potential functional roles. We identified two promising genes in the DNA repair enzyme DNA polymerase beta and in the neuroendocrine transcription factor NKX2.2. Specific inhibition of DNA polymerase beta reduced the numbers of cells surviving treatment with etoposide and increased the amount of DNA damage in cells. Conversely, stable overexpression of NKX2.2 increased cell survival in response to etoposide in SCLC cell lines. Consistent with these findings, we found that an absence of nuclear staining for NKX2.2 in SCLC primary tumors was an independent predictor of improved outcomes in chemotherapy-treated patients. Taken together, our findings justify future prospective studies to confirm the roles of these molecules in mediating chemotherapy resistance in SCLC. PMID- 21642374 TI - Combined gene expression profiling and RNAi screening in clear cell renal cell carcinoma identify PLK1 and other therapeutic kinase targets. AB - In recent years, several molecularly targeted therapies have been approved for clear cell renal cell carcinoma (ccRCC), a highly aggressive cancer. Although these therapies significantly extend overall survival, nearly all patients with advanced ccRCC eventually succumb to the disease. To identify other molecular targets, we profiled gene expression in 90 ccRCC patient specimens for which tumor grade information was available. Gene set enrichment analysis indicated that cell-cycle-related genes, in particular, Polo-like kinase 1 (PLK1), were associated with disease aggressiveness. We also carried out RNAi screening to identify kinases and phosphatases that when inhibited could prevent cell proliferation. As expected, RNAi-mediated knockdown of PLK1 and other cell-cycle kinases was sufficient to suppress ccRCC cell proliferation. The association of PLK1 in both disease aggression and in vitro growth prompted us to examine the effects of a small-molecule inhibitor of PLK1, BI 2536, in ccRCC cell lines. BI 2536 inhibited the proliferation of ccRCC cell lines at concentrations required to inhibit PLK1 kinase activity, and sustained inhibition of PLK1 by BI 2536 led to dramatic regression of ccRCC xenograft tumors in vivo. Taken together, these findings highlight PLK1 as a rational therapeutic target for ccRCC. PMID- 21642376 TI - Effects of hypothyroidism on the structure and mechanical properties of bone in the ovine fetus. AB - Thyroid hormones are important for normal bone growth and development in postnatal life. However, little is known about the role of thyroid hormones in the control of bone development in the fetus. Using computed tomography and mechanical testing, the structure and strength of metatarsal bones were measured in sheep fetuses in which thyroid hormone levels were altered by thyroidectomy or adrenalectomy. In intact fetuses, plasma concentrations of total calcium and the degradation products of C-terminal telopeptides of type I collagen increased between 100 and 144 days of gestation (term 145+/-2 days), in association with various indices of bone growth and development. Thyroid hormone deficiency induced by thyroidectomy at 105-110 days of gestation caused growth retardation of the fetus and significant changes in metatarsal bone structure and strength when analyzed at both 130 and 144 days of gestation. In hypothyroid fetuses, trabecular bone was stronger with thicker, more closely spaced trabeculae, despite lower bone mineral density. Plasma osteocalcin was reduced by fetal thyroidectomy. Removal of the fetal adrenal gland at 115-120 days of gestation, and prevention of the prepartum rises in cortisol and triiodothyronine, had no effect on bodyweight, limb lengths, metatarsal bone structure or strength, or circulating markers of bone metabolism in the fetuses studied near term. This study demonstrates that hypothyroidism in utero has significant effects on the structure and strength of bone, with different consequences for cortical and trabecular bone. PMID- 21642375 TI - Mechanical stiffness grades metastatic potential in patient tumor cells and in cancer cell lines. AB - Cancer cells are defined by their ability to invade through the basement membrane, a critical step during metastasis. While increased secretion of proteases, which facilitates degradation of the basement membrane, and alterations in the cytoskeletal architecture of cancer cells have been previously studied, the contribution of the mechanical properties of cells in invasion is unclear. Here, we applied a magnetic tweezer system to establish that stiffness of patient tumor cells and cancer cell lines inversely correlates with migration and invasion through three-dimensional basement membranes, a correlation known as a power law. We found that cancer cells with the highest migratory and invasive potential are five times less stiff than cells with the lowest migration and invasion potential. Moreover, decreasing cell stiffness by pharmacologic inhibition of myosin II increases invasiveness, whereas increasing cell stiffness by restoring expression of the metastasis suppressor TbetaRIII/betaglycan decreases invasiveness. These findings are the first demonstration of the power law relation between the stiffness and the invasiveness of cancer cells and show that mechanical phenotypes can be used to grade the metastatic potential of cell populations with the potential for single cell grading. The measurement of a mechanical phenotype, taking minutes rather than hours needed for invasion assays, is promising as a quantitative diagnostic method and as a discovery tool for therapeutics. By showing that altering stiffness predictably alters invasiveness, our results indicate that pathways regulating these mechanical phenotypes are novel targets for molecular therapy of cancer. PMID- 21642377 TI - Mutational analysis of GCMB, a parathyroid-specific transcription factor, in parathyroid adenoma of primary hyperparathyroidism. AB - Sporadic primary hyperparathyroidism (PHPT), one of the most common endocrine disorders, is characterized by hypercalcemia and elevated PTH levels. The majority of cases are caused by a benign parathyroid adenoma, but somatic or de novo germ-line mutations that lead to adenoma formation have only been identified in few glands. GCMB is a parathyroid-specific transcription factor, which causes hypoparathyroidism when inactivated on both parental alleles or when a dominant negative, heterozygous mutation is present. It is overexpressed in some parathyroid adenomas, and we therefore tested the hypothesis that GCMB mutations can be a cause of parathyroid adenomas. Nucleotide sequence analysis was performed on all coding exons and exon-intron borders of GCMB in 30 sporadic parathyroid adenomas and we identified several known polymorphisms that were either heterozygous or homozygous. In addition, one of the 30 investigated glands revealed a novel heterozygous missense mutation, c.1144G>A, which introduced methionine at position 382 for valine (V382M), a conserved amino acid residue. Western blot analysis using mutant GCMB (GCMB-V382M) from lysates of transiently transfected DF-1 fibroblasts, luciferase assays using extracts from these cells, and electrophoretic mobility assays failed to reveal differences between wild type and mutant GCMB in expression level, transactivational capacity, and DNA binding ability. Furthermore, pulse-chase experiments demonstrated no difference in half-life of wild-type and mutant protein. We conclude that mutations in the transcription factor GCMB do not seem to play a major role in the pathogenesis of PHPT. PMID- 21642379 TI - The skeleton: a multi-functional complex organ: the growth plate chondrocyte and endochondral ossification. AB - Endochondral ossification is the process that results in both the replacement of the embryonic cartilaginous skeleton during organogenesis and the growth of long bones until adult height is achieved. Chondrocytes play a central role in this process, contributing to longitudinal growth through a combination of proliferation, extracellular matrix (ECM) secretion and hypertrophy. Terminally differentiated hypertrophic chondrocytes then die, allowing the invasion of a mixture of cells that collectively replace the cartilage tissue with bone tissue. The behaviour of growth plate chondrocytes is tightly regulated at all stages of endochondral ossification by a complex network of interactions between circulating hormones (including GH and thyroid hormone), locally produced growth factors (including Indian hedgehog, WNTs, bone morphogenetic proteins and fibroblast growth factors) and the components of the ECM secreted by the chondrocytes (including collagens, proteoglycans, thrombospondins and matrilins). In turn, chondrocytes secrete factors that regulate the behaviour of the invading bone cells, including vascular endothelial growth factor and receptor activator of NFkappaB ligand. This review discusses how the growth plate chondrocyte contributes to endochondral ossification, with some emphasis on recent advances. PMID- 21642378 TI - eNOS activation and NO function: structural motifs responsible for the posttranslational control of endothelial nitric oxide synthase activity. AB - Rather than being a constitutive enzyme as was first suggested, endothelial nitric oxide synthase (eNOS) is dynamically regulated at the transcriptional, posttranscriptional, and posttranslational levels. This review will focus on how changes in eNOS function are conferred by various posttranslational modifications. The latest knowledge regarding eNOS targeting to the plasma membrane will be discussed as the role of protein phosphorylation as a modulator of catalytic activity. Furthermore, new data are presented that provide novel insights into how disruption of the eNOS dimer prevents eNOS uncoupling and the production of superoxide under conditions of elevated oxidative stress and identifies a novel regulatory region we have termed the 'flexible arm'. PMID- 21642380 TI - Molecular chaperones as a common set of proteins that regulate the invasion phenotype of head and neck cancer. AB - PURPOSE: The goal of this study was to establish a common set of molecules that regulate cell invasion in head and neck cancer (HNC). EXPERIMENTAL DESIGN: Five invasive sublines derived from HNC cell lines were established using the Matrigel selection method. Proteomic technology, MetaCore algorithm, and reverse transcriptase-PCR methods were used to search for molecules that contribute to the invasion phenotype. Cellular functional analyses and clinical association studies were applied to examine the significance of the molecules. RESULTS: Fifty two proteins were identified in more than two of the four independent proteomic experiments, including 10 (19%) molecular chaperones. Seven chaperones were confirmed to be differentially expressed in five sublines, Hsp90alpha, Hsp90beta, Hsp90-B1/Gp96, Hsp70-A5/Grp78, and HYOU1, that upregulate, whereas Hsp60 and glucosidase-alpha neutral AB (GANAB) downregulate. Four molecules were further investigated. In all cell lines, knockdown of Hsp60 or GANAB and silencing of Gp96 or Grp78 considerably enhanced or reduced cell migration and invasion, respectively. Clinical association studies consistently revealed that low levels of Hsp60 or GANAB and high levels of Gp96 or Grp78 are significantly associated with advanced cancer (P < 0.001 to P = 0.047, respectively, for the four molecules) and poor survival (P < 0.001 to P = 0.025, respectively, for the four molecules). CONCLUSION: Our study defined molecular chaperones as a common set of proteins that regulate the invasion phenotype of HNC. Loss of the tumor suppression function of Hsp60 or GANAB and acquisition of the oncogenic function of Gp96 or Grp78 contribute to aggressive cancers. These molecules may serve as prognostic markers and targets for cancer drug development. PMID- 21642381 TI - The efficacy of IGF-I receptor monoclonal antibody against human gastrointestinal carcinomas is independent of k-ras mutation status. AB - PURPOSE: Insulin-like growth factor (IGF)-I receptor (IGF-IR) signaling is required for carcinogenicity and proliferation of gastrointestinal cancers. We have previously shown successful targeting therapy for colorectal, pancreatic, gastric, and esophageal carcinomas using recombinant adenoviruses expressing dominant negative IGF-IR. Mutation in k-ras is one of key factors in gastrointestinal cancers. In this study, we sought to evaluate the effect of a new monoclonal antibody for IGF-IR, figitumumab (CP-751,871), on the progression of human gastrointestinal carcinomas with/without k-ras mutation. EXPERIMENTAL DESIGN: We assessed the effect of figitumumab on signal transduction, proliferation, and survival in six gastrointestinal cancer cell lines with/without k-ras mutation, including colorectal and pancreatic adenocarcinoma, esophageal squamous cell carcinoma, and hepatoma. Combination effects of figitumumab and chemotherapy were also studied. Then figitumumab was evaluated in the treatment of xenografts in nude mice. RESULTS: Figitumumab blocked autophosphorylation of IGF-IR and its downstream signals. The antibody suppressed proliferation and tumorigenicity in all cell lines. Figitumumab inhibited survival by itself and up-regulated chemotherapy (5-FU and gemcitabine) induced apoptosis. Moreover, the combination of this agent and chemotherapy was effective against tumors in mice. The effect of figitumumab was not influenced by the mutation status of k-ras. Figitumumab reduced expression of IGF-IR but not insulin receptor in these xenografted tumors. The drug did not affect murine body weight or blood concentrations of glucose, insulin, IGF binding protein 3, and growth hormone. CONCLUSIONS: IGF-IR might be a good molecular therapeutic target and figitumumab may thus have therapeutic value in human gastrointestinal malignancies even in the presence of k-ras mutations. PMID- 21642382 TI - Merkel cell polyomavirus infection, large T antigen, retinoblastoma protein and outcome in Merkel cell carcinoma. AB - PURPOSE: Merkel cell carcinoma (MCC) is rare skin cancer that is often associated with Merkel cell polyomavirus (MCPyV) infection. Polyomaviruses repress tumor suppressor proteins, thus influencing cell-cycle progression, but the effect of MCPyV on the key cell-cycle regulating proteins is poorly understood. EXPERIMENTAL DESIGN: We evaluated expression of the MCPyV large T-antigen (LTA), Ki-67, and the key putative tumor suppressor proteins, the retinoblastoma protein (RB and phospho-RB) and p53, and their regulatory proteins (cyclin D1, cyclin E, p16, p21, p27, and MDM2) by using immunohistochemistry from tumors of 91 MCC patients identified from a population-based nationwide cohort. Tumor MCPyV DNA was measured by using quantitative PCR, and TP53 mutations were identified with sequencing. RESULTS: MCPyV LTA expression was strongly associated with presence of MCPyV DNA in tumor, and it was almost invariably associated with tumor RB expression (P < 0.0001 for both comparisons). Both MCC LTA and RB expression were strongly associated with favorable MCC-specific and overall survival in univariable analyses (P <= 0.01 for all four analyses). Presence of MCPyV LTA was also associated with the female gender, the intermediate type of tumor histology, location of the tumor in a limb, cell proliferation rate, and absence of p53 expression. TP53 mutations were detected only in MCPyV DNA-negative tumors. CONCLUSIONS: MCPyV DNA-positive MCC has several clinical and molecular features that differ from MCPyV DNA-negative cancers. MCPyV-associated MCCs express RB, but may not harbor TP53 mutations. These findings provide further support that MCPyV causes the majority of MCCs. PMID- 21642383 TI - Sperm acrosin is responsible for the sperm binding to the egg envelope during fertilization in Japanese quail (Coturnix japonica). AB - An antibody library against quail sperm plasma membrane components was established and a mAb, which strongly inhibits sperm perforations of the perivitelline membrane (PVM) was obtained from the library. The antigen molecule of the mAb showed an apparent molecular weight of 45 kDa, and was distributed both on the surface and in the acrosomal matrix of the sperm head. Periodate oxidation revealed that the epitope of the antigen includes a sugar moiety. Tandem mass spectrometry analysis of the antigen revealed that the mAb recognizes sperm acrosin. When sodium dodecyl sulfate-solubilized PVM immobilized on a polyvinylidene difluoride membrane was incubated with sperm plasma membrane lysates, the sperm acrosin was detected on the PVM immobilized on the membrane, indicating that the sperm acrosin interacts with the components of PVM. Indeed, the mAb effectively inhibited the binding of acrosome-intact sperm to the PVM. These results indicate that the 45 kDa sperm acrosin is involved in the binding of sperm to the PVM in fertilization of Japanese quail. PMID- 21642385 TI - Association of an SNP with intrathymic transcription of TSHR and Graves' disease: a role for defective thymic tolerance. AB - Graves' disease (GD) is the paradigm of an anti-receptor autoimmune disease, with agonistic auto-antibodies against the thyrotropin receptor (TSHR-thyroid stimulating hormone receptor) being the underlying pathogenic effector mechanism. The TSHR belongs to the category of tissue-restricted antigens, which are promiscuously expressed in the thymus and thereby induce central T cell tolerance. In order to understand the association between TSHR gene polymorphisms and GD, we tested the hypothesis that TSHR gene variants affect susceptibility to GD by influencing levels of TSHR transcription in the thymus. We show that thymic glands from non-autoimmune donors homozygous for the rs179247 SNP predisposing allele of TSHR had significantly fewer TSHR mRNA transcripts than carriers of the protective allele. In addition, in heterozygous individuals, the TSHR predisposing allele was expressed at a lower level than the protective one as demonstrated by allele-specific transcript quantification. This unbalanced allelic expression was detectable in both thymic epithelial cells and thymocytes. Since the level of self-antigen expression is known to influence the threshold of central tolerance, these results are compatible with the notion that defective central tolerance contributes to the pathogenesis of GD, a scenario already implicated in type 1 diabetes mellitus, myasthenia gravis and autoimmune myocarditis. PMID- 21642384 TI - Glucose transporters in the uterus: an analysis of tissue distribution and proposed physiological roles. AB - Facilitative glucose transport molecules (glucose transporters, GLUTs) are responsible for glucose transport across cellular membranes. Of the 14 family members, expression of nine has been reported in the murine uterus and seven in the human uterus. Some studies reveal that adequate glucose uptake and metabolism are essential for the proper differentiation of the uterine endometrium toward a receptive state capable of supporting embryo implantation. However, the mechanistic role of GLUTs in endometrial function remains poorly understood. This review aims to present the current knowledge about GLUT expression in the uterus and distribution among the different cell types within the endometrium. In addition, it analyzes the available data in the context of roles GLUTs may play in normal uterine physiology as well as the pathological conditions of infertility, endometrial cancer, and polycystic ovarian syndrome. PMID- 21642387 TI - Expression of the osteoarthritis-associated gene GDF5 is modulated epigenetically by DNA methylation. AB - GDF5 is involved in synovial joint development, maintenance and repair, and the rs143383 C/T single nucleotide polymorphism (SNP) located in the 5'UTR of GDF5 is associated, at the genome-wide significance level, with osteoarthritis susceptibility, and with other musculoskeletal phenotypes including height, congenital hip dysplasia and Achilles tendinopathy. There is a significant reduction in the expression of the disease-associated T allele relative to the C allele in synovial joint tissues, an effect influenced by a second SNP (rs143384, C/T) also within the 5'UTR. The differential allelic expression (DAE) imbalance of the C and T alleles of rs143383 varies intra- and inter-individually, suggesting that DAE may be modulated epigenetically. The C alleles of both SNPs form CpG dinucleotides that are potentially amenable to regulation by methylation. Here, we have examined whether DNA methylation regulates GDF5 expression and the allelic imbalance caused by rs143383. We observed methylation of the GDF5 promoter and 5'UTR in cell lines and joint tissues, with demethylation correlating with increased GDF5 expression. The CpG sites created by the C alleles at rs143383 and rs143384 were variably methylated, and treatment of a heterozygous cell line with a demethylating agent further increased the allelic expression imbalance between the C and T alleles. This demonstrates that the genetic effect of the rs143383 SNP on GDF5 expression is modulated epigenetically by DNA methylation. The variability in DAE of rs143383 is therefore partly accounted for by differences in DNA methylation that could influence the penetrance of this allele in susceptibility to common musculoskeletal diseases. PMID- 21642386 TI - Alpha-synuclein functions in the nucleus to protect against hydroxyurea-induced replication stress in yeast. AB - Hydroxyurea (HU) inhibits ribonucleotide reductase (RNR), which catalyzes the rate-limiting synthesis of deoxyribonucleotides for DNA replication. HU is used to treat HIV, sickle-cell anemia and some cancers. We found that, compared with vector control cells, low levels of alpha-synuclein (alpha-syn) protect S. cerevisiae cells from the growth inhibition and reactive oxygen species (ROS) accumulation induced by HU. Analysis of this effect using different alpha-syn mutants revealed that the alpha-syn protein functions in the nucleus and not the cytoplasm to modulate S-phase checkpoint responses: alpha-syn up-regulates histone acetylation and RNR levels, maintains helicase minichromosome maintenance protein complexes (Mcm2-7) on chromatin and inhibits HU-induced ROS accumulation. Strikingly, when residues 2-10 or 96-140 are deleted, this protective function of alpha-syn in the nucleus is abolished. Understanding the mechanism by which alpha syn protects against HU could expand our knowledge of the normal function of this neuronal protein. PMID- 21642388 TI - Classifying tissue samples from measurements on cells with within-class tissue sample heterogeneity. AB - We consider here the problem of classifying a macro-level object based on measurements of embedded (micro-level) observations within each object, for example, classifying a patient based on measurements on a collection of a random number of their cells. Classification problems with this hierarchical, nested structure have not received the same statistical understanding as the general classification problem. Some heuristic approaches have been developed and a few authors have proposed formal statistical models. We focus on the problem where heterogeneity exists between the macro-level objects within a class. We propose a model-based statistical methodology that models the log-odds of the macro-level object belonging to a class using a latent-class variable model to account for this heterogeneity. The latent classes are estimated by clustering the macro level object density estimates. We apply this method to the detection of patients with cervical neoplasia based on quantitative cytology measurements on cells in a Papanicolaou smear. Quantitative cytology is much cheaper and potentially can take less time than the current standard of care. The results show that the automated quantitative cytology using the proposed method is roughly equivalent to clinical cytopathology and shows significant improvement over a statistical model that does not account for the heterogeneity of the data. PMID- 21642389 TI - A fused lasso latent feature model for analyzing multi-sample aCGH data. AB - Array-based comparative genomic hybridization (aCGH) enables the measurement of DNA copy number across thousands of locations in a genome. The main goals of analyzing aCGH data are to identify the regions of copy number variation (CNV) and to quantify the amount of CNV. Although there are many methods for analyzing single-sample aCGH data, the analysis of multi-sample aCGH data is a relatively new area of research. Further, many of the current approaches for analyzing multi sample aCGH data do not appropriately utilize the additional information present in the multiple samples. We propose a procedure called the Fused Lasso Latent Feature Model (FLLat) that provides a statistical framework for modeling multi sample aCGH data and identifying regions of CNV. The procedure involves modeling each sample of aCGH data as a weighted sum of a fixed number of features. Regions of CNV are then identified through an application of the fused lasso penalty to each feature. Some simulation analyses show that FLLat outperforms single-sample methods when the simulated samples share common information. We also propose a method for estimating the false discovery rate. An analysis of an aCGH data set obtained from human breast tumors, focusing on chromosomes 8 and 17, shows that FLLat and Significance Testing of Aberrant Copy number (an alternative, existing approach) identify similar regions of CNV that are consistent with previous findings. However, through the estimated features and their corresponding weights, FLLat is further able to discern specific relationships between the samples, for example, identifying 3 distinct groups of samples based on their patterns of CNV for chromosome 17. PMID- 21642390 TI - Role of CXCL13-CXCR5 crosstalk between malignant neuroblastoma cells and Schwannian stromal cells in neuroblastic tumors. AB - Neuroblastoma is a stroma-poor (SP) aggressive pediatric cancer belonging to neuroblastic tumors, also including ganglioneuroblastoma and ganglioneuroma, two stroma-rich (SR) less aggressive tumors. Our previous gene-expression profiling analysis showed a different CXCL13 mRNA expression between SP and SR tumors. Therefore, we studied 13 SP and 13 SR tumors by reverse transcription quantitative real-time PCR (RT-qPCR) and we found that CXCR5b was more expressed in SP than in SR and CXCL13 was predominantly expressed in SR tumors. Then, we isolated neuroblastic and Schwannian stromal cells by laser capture microdissection and we found that malignant neuroblasts express CXCR5b mRNA, whereas Schwannian stromal cells express CXCL13. Immunohistochemistry confirmed that stroma expresses CXCL13 but not CXCR5. To better understand the role of CXCL13 and CXCR5 in neuroblastic tumors we studied 11 neuroblastoma cell lines and we detected a heterogeneous expression of CXCL13 and CXCR5b. Interestingly, we found that only CXCR5b splice variant was expressed in both tumors and neuroblastoma lines, whereas CXCR5a was never detected. Moreover, we found that neuroblastoma cells expressing CXCR5 receptor migrate toward a source of recombinant CXCL13. Lastly, neuroblastoma cells induced to glial cell differentiation expressed CXCL13 mRNA and protein. The chemokine released in the culture medium was able to stimulate chemotaxis of LA1-5S neuroblastoma cells. Collectively, our data suggest that CXCL13 produced by stromal cells may contribute to the generation of an environment in which the malignant neuroblasts are retained, thus limiting the possible development of metastases in patients with SR tumor. PMID- 21642391 TI - Susceptibility to tuberculosis is associated with TLR1 polymorphisms resulting in a lack of TLR1 cell surface expression. AB - Human TLR1 plays an important role in host defense against Mycobacterium tuberculosis. Our aim was to analyze the association of the loss of TLR1 surface expression and TLR1 SNPs with susceptibility to TB. TLR1neg and TLR1pos cells from healthy individuals were identified by flow cytometry and compared by sequencing. TLR1 expression was measured using quantitative real-time PCR and immunoblotting. TLR1 SNP analyses of healthy individuals and TB patients from EU C and Ghana were performed, and association of the TLR1 genotypes with increased risk of developing TB was statistically evaluated. Lack of TLR1 surface expression accompanied by impaired function was strongly associated with TLR1 SNP G743A. Genotyping of EU-C controls and TB patients revealed an association of TLR1 743A/1805G alleles [OR 2.37 (95% CI 1.13, 4.93), P=0.0219; OR 2.74 (95% CI 1.26, 6.05), P=0.0059] as well as TLR1neg 743AA/1805GG versus TLR1pos genotypes 743AG/1805TG [OR 4.98 (95% CI 1.64, 15.15), P=0.0034; OR 5.70 (95% CI 1.69, 20.35), P=0.0015] and 743AG + GG/1805TG + TT [OR 3.54 (95% CI 1.29, 9.90), P=0.0086; OR 4.17 (95% CI 1.52, 11.67), P=0.0025] with increased susceptibility to TB. No association of G743A with TB was found in Ghana as a result of a low frequency of genotype 743AA. Our data gain new insights in the role of TLR1 in M. tuberculosis defense and provide the first evidence that TLR1 variants are associated with susceptibility to TB in a low-incidence country. PMID- 21642392 TI - Interaction of N-(2-hydroxypropyl)methacrylamide copolymer-doxorubicin conjugates with human liver microsomal cytochromes P450: comparison with free doxorubicin. AB - Interaction of nine forms of human hepatic cytochromes P450 (CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, and CYP3A4) with two N-(2 hydroxypropyl)methacrylamide (HPMA) copolymer-based doxorubicin (DOX) conjugates designed for passive tumor targeting was studied using pooled human microsomes. The compounds used in this study were two high-molecular-weight HPMA copolymers bearing doxorubicin attached to the polymeric carrier by 1) hydrazone bond enabling intracellular pH-controlled drug release or 2) amide bond through enzymatically cleavable tetrapeptide GlyPheLeuGly spacer. Both polymeric conjugates differing in mechanism of their antitumor activity and the free doxorubicin as the control were tested for potential inhibition activity. Among nine cytochrome P450 forms studied, no HPMA copolymer with bound DOX caused an inhibition of potential clinical significance. The extent of inhibition of enzymatic activities of the cytochrome P450 forms studied was negligible with the exception of CYP2B6 and was apparently caused by DOX as no inhibition was observed with polymers alone, and the extent of inhibition by the complex corresponded to this of the free DOX at the same concentration. In conclusion, the polymers and their conjugates with DOX seem to be relatively safe, at least in this respect, i.e., of inhibition of the liver microsomal drug-metabolizing enzymes. PMID- 21642393 TI - Functional and structural relevance of conserved positively charged lysine residues in organic anion transporting polypeptide 1B3. AB - The human organic anion transporting polypeptide 1B3 (OATP1B3), located in the basolateral membrane of hepatocytes, mediates the uptake of endogenous substrates such as taurocholate and drugs from blood into hepatocytes. The transport activity of OATP1B3 is influenced by positively charged amino acids, which are facing the central pore. Molecular modeling was performed to select conserved positively charged amino acids, which may influence transport activity and anchoring of OATP1B3 in the plasma membrane. The modeling revealed that Lys361 faces the pore, and Lys399 is oriented to the plasma membrane. Therefore, the mutants L361>A, L361>R, L399>A, and L399>R were generated using site-directed mutagenesis to investigate the impact of the positive charges on transport activity and anchoring in the membrane. Transport kinetic analyses for the substrates sulfobromophthalein and taurocholate showed a loss of function for the L361>A mutant, whereas the transport activity was maintained by the L361>R mutant, indicating that the positive charge at position 361 is important for transport activity of OATP1B3. Comparative modeling with OATP1A2 and OATP2B1 revealed that the pore size around this lysine residue is larger in OATP1A2 and smaller in OATP2B1 compared with OATP1B3, which could be related to the respective substrate spectra. Cell surface expression of L399>A and L399>R was decreased to 16 and 72% compared with wild-type OATP1B3 (p < 0.001), respectively, indicating that the positive charge of lysine at position 399 is necessary for an unimpaired cell surface expression. Furthermore, we provide a summary of amino acids, which influence the transport activity of OATP1B3. PMID- 21642394 TI - Calcium/calmodulin-dependent kinase II mediates the phosphorylation and activation of NADPH oxidase 5. AB - Excessive synthesis of reactive oxygen species contributes to the pathology of many human diseases and originates from changes in the expression and posttranslational regulation of the transmembrane NADPH oxidases (Noxes). Nox5 is a novel Nox isoform whose activity is regulated by intracellular calcium levels. We have reported that the activity and calcium-sensitivity of Nox5 can also be modulated by direct phosphorylation. However, the kinases that phosphorylate Nox5 have not been identified, and thus, the goal of this study was to determine whether calcium-activated kinases such as calcium/calmodulin-dependent kinase II (CAMKII) are involved. We found that Nox5 activity in bovine aortic endothelial cells was suppressed by two doses of the CAMKII inhibitor 2-(N-[2-hydroxyethyl]) N-(4-methoxybenzenesulfonyl)amino-N-(4-chlorocinnamyl)-N-methylamine (KN-93). In cotransfected COS-7 cells, wild-type and constitutively active CAMKII, but not a dominant-negative, robustly increased basal Nox5 activity. The ability of CAMKII to increase Nox5 activity was also observed with fixed calcium concentrations in an isolated enzyme activity assay. CAMKII did not elevate intracellular calcium or activate other Nox enzymes. In vitro phosphorylation assays revealed that CAMKII can directly phosphorylate Nox5 on Thr494 and Ser498 as detected by phosphorylation state-specific antibodies. Mass spectrometry (MS) analysis revealed the phosphorylation of additional, novel sites at Ser475, Ser502, and Ser675. Of these phosphorylation sites, mutation of only Ser475 to alanine prevented CAMKII-induced increases in Nox5 activity. The ability of CAMKIIalpha to phosphorylate Ser475 in intact cells was supported by the binding of Nox5 to phosphoprotein-affinity columns and via MS/MS analysis. Together, these results suggest that CAMKII can positively regulate Nox5 activity via the phosphorylation of Ser475. PMID- 21642395 TI - Decay of bacterial pathogens, fecal indicators, and real-time quantitative PCR genetic markers in manure-amended soils. AB - This study examined persistence and decay of bacterial pathogens, fecal indicator bacteria (FIB), and emerging real-time quantitative PCR (qPCR) genetic markers for rapid detection of fecal pollution in manure-amended agricultural soils. Known concentrations of transformed green fluorescent protein-expressing Escherichia coli O157:H7/pZs and red fluorescent protein-expressing Salmonella enterica serovar Typhimurium/pDs were added to laboratory-scale manure-amended soil microcosms with moisture contents of 60% or 80% field capacity and incubated at temperatures of -20 degrees C, 10 degrees C, or 25 degrees C for 120 days. A two-stage first-order decay model was used to determine stage 1 and stage 2 first order decay rate coefficients and transition times for each organism and qPCR genetic marker in each treatment. Genetic markers for FIB (Enterococcus spp., E. coli, and Bacteroidales) exhibited decay rate coefficients similar to that of E. coli O157:H7/pZs but not of S. enterica serovar Typhimurium/pDs and persisted at detectable levels longer than both pathogens. Concentrations of these two bacterial pathogens, their counterpart qPCR genetic markers (stx1 and ttrRSBCA, respectively), and FIB genetic markers were also correlated (r = 0.528 to 0.745). This suggests that these qPCR genetic markers may be reliable conservative surrogates for monitoring fecal pollution from manure-amended land. Host associated qPCR genetic markers for microbial source tracking decayed rapidly to nondetectable concentrations, long before FIB, Salmonella enterica serovar Typhimurium/pDs, and E. coli O157:H7/pZs. Although good indicators of point source or recent nonpoint source fecal contamination events, these host associated qPCR genetic markers may not be reliable indicators of nonpoint source fecal contamination events that occur weeks following manure application on land. PMID- 21642396 TI - Global gene expression profile for swarming Bacillus cereus bacteria. AB - Bacillus cereus can use swarming to move over and colonize solid surfaces in different environments. This kind of motility is a collective behavior accompanied by the production of long and hyperflagellate swarm cells. In this study, the genome-wide transcriptional response of B. cereus ATCC 14579 during swarming was analyzed. Swarming was shown to trigger the differential expression (>2-fold change) of 118 genes. Downregulated genes included those required for basic cellular metabolism. In accordance with the hyperflagellate phenotype of the swarm cell, genes encoding flagellin were overexpressed. Some genes associated with K(+) transport, phBC6A51 phage genes, and the binding component of the enterotoxin hemolysin BL (HBL) were also induced. Quantitative reverse transcription-PCR (qRT-PCR) experiments indicated an almost 2-fold upregulation of the entire hbl operon during swarming. Finally, BC1435 and BC1436, orthologs of liaI-liaH that are known to be involved in the resistance of Bacillus subtilis to daptomycin, were upregulated under swarming conditions. Accordingly, phenotypic assays showed reduced susceptibility of swarming B. cereus cells to daptomycin, and P(spac)-induced hyper-expression of these genes in liquid medium highlighted the role of BC1435 and BC1436 in the response of B. cereus to daptomycin. PMID- 21642397 TI - Influence of environmental and genetic factors linked to celiac disease risk on infant gut colonization by Bacteroides species. AB - Celiac disease (CD) is an immune-mediated enteropathy involving genetic and environmental factors whose interaction might influence disease risk. The aim of this study was to determine the effects of milk-feeding practices and the HLA-DQ genotype on intestinal colonization of Bacteroides species in infants at risk of CD development. This study included 75 full-term newborns with at least one first degree relative suffering from CD. Infants were classified according to milk feeding practice (breast-feeding or formula feeding) and HLA-DQ genotype (high or low genetic risk). Stools were analyzed at 7 days, 1 month, and 4 months by PCR and denaturing gradient gel electrophoresis (DGGE). The Bacteroides species diversity index was higher in formula-fed infants than in breast-fed infants. Breast-fed infants showed a higher prevalence of Bacteroides uniformis at 1 and 4 months of age, while formula-fed infants had a higher prevalence of B. intestinalis at all sampling times, of B. caccae at 7 days and 4 months, and of B. plebeius at 4 months. Infants with high genetic risk showed a higher prevalence of B. vulgatus, while those with low genetic risk showed a higher prevalence of B. ovatus, B. plebeius, and B. uniformis. Among breast-fed infants, the prevalence of B. uniformis was higher in those with low genetic risk than in those with high genetic risk. Among formula-fed infants, the prevalence of B. ovatus and B. plebeius was increased in those with low genetic risk, while the prevalence of B. vulgatus was higher in those with high genetic risk. The results indicate that both the type of milk feeding and the HLA-DQ genotype influence the colonization process of Bacteroides species, and possibly the disease risk. PMID- 21642398 TI - Role of nitric oxide and flavohemoglobin homolog genes in Aspergillus nidulans sexual development and mycotoxin production. AB - Flavohemoglobins are widely distributed in both prokaryotes and eukaryotes. These proteins are involved in reducing nitric oxide levels. Deletion of the Aspergillus nidulans flavohemoglobin gene fhbA induced sexual development and decreased sterigmatocystin production. Supplementation with a nitric oxide releasing compound promoted cleistothecial formation and increased nsdD and steA expression, indicating that nitric oxide induces sexual development. This is the first study on the effect of nitric oxide on morphogenesis and secondary metabolism in fungi. PMID- 21642399 TI - Statistical analysis of long- and short-range forces involved in bacterial adhesion to substratum surfaces as measured using atomic force microscopy. AB - Surface thermodynamic analyses of microbial adhesion using measured contact angles on solid substrata and microbial cell surfaces are widely employed to determine the nature of the adhesion forces, i.e., the interplay between Lifshitz van der Waals and acid-base forces. While surface thermodynamic analyses are often viewed critically, atomic force microscopy (AFM) can also provide information on the nature of the adhesion forces by means of Poisson analysis of the measured forces. This review first presents a description of Poisson analysis and its underlying assumptions. The data available from the literature for different combinations of bacterial strains and substrata are then summarized, leading to the conclusion that bacterial adhesion to surfaces is generally dominated by short-range, attractive acid-base interactions, in combination with long-range, weaker Lifshitz-van der Waals forces. This is in line with the findings of surface thermodynamic analyses of bacterial adhesion. Comparison with single-molecule ligand-receptor forces from the literature suggests that the short-range-force contribution from Poisson analysis involves a discrete adhesive bacterial cell surface site rather than a single molecular force. The adhesion force arising from these cell surface sites and the number of sites available may differ from strain to strain. Force spectroscopy, however, involves the tedious task of identifying the minor peaks in the AFM retraction force-distance curve. This step can be avoided by carrying out Poisson analysis on the work of adhesion, which can also be derived from retraction force-distance curves. This newly proposed way of performing Poisson analysis confirms that multiple molecular bonds, rather than a single molecular bond, contribute to a discrete adhesive bacterial cell surface site. PMID- 21642400 TI - Increased transfer of a multidrug resistance plasmid in Escherichia coli biofilms at the air-liquid interface. AB - Although biofilms represent a common bacterial lifestyle in clinically and environmentally important habitats, there is scant information on the extent of gene transfer in these spatially structured populations. The objective of this study was to gain insight into factors that affect transfer of the promiscuous multidrug resistance plasmid pB10 in Escherichia coli biofilms. Biofilms were grown in different experimental settings, and plasmid transfer was monitored using laser scanning confocal microscopy and plate counting. In closed flow cells, plasmid transfer in surface-attached submerged biofilms was negligible. In contrast, a high plasmid transfer efficiency was observed in a biofilm floating at the air-liquid interface in an open flow cell with low flow rates. A vertical flow cell and a batch culture biofilm reactor were then used to detect plasmid transfer at different depths away from the air-liquid interface. Extensive plasmid transfer occurred only in a narrow zone near that interface. The much lower transfer frequencies in the lower zones coincided with rapidly decreasing oxygen concentrations. However, when an E. coli csrA mutant was used as the recipient, a thick biofilm was obtained at all depths, and plasmid transfer occurred at similar frequencies throughout. These results and data from separate aerobic and anaerobic matings suggest that oxygen can affect IncP-1 plasmid transfer efficiency, not only directly but also indirectly, through influencing population densities and therefore colocalization of donors and recipients. In conclusion, the air-liquid interface can be a hot spot for plasmid-mediated gene transfer due to high densities of juxtaposed donor and recipient cells. PMID- 21642401 TI - Involvement of multiple loci in quorum quenching of autoinducer I molecules in the nitrogen-fixing symbiont Rhizobium (Sinorhizobium) sp. strain NGR234. AB - Rhizobium sp. strain NGR234 is a unique alphaproteobacterium (order Rhizobiales) that forms nitrogen-fixing nodules with more legumes than any other microsymbiont. Since we have previously described the complete genome sequence of NGR234, we now report on a genome-wide functional analysis of the genes and enzymes involved in autoinducer I hydrolysis in this microbe. Altogether we identified five cosmid clones that repeatedly gave a positive result in our function-based approach for the detection of autoinducer I hydrolase genes. Of these five cosmid clones, two were located on pNGR234b and three were on cNGR234. Subcloning and in vitro mutagenesis in combination with BLAST analyses identified the corresponding open reading frames (ORFs) of all cosmid clones: dlhR, qsdR1, qsdR2, aldR, and hitR-hydR. Analyses of recombinant DlhR and QsdR1 proteins by using high-performance liquid chromatography-mass spectrometry (HPLC-MS) demonstrate that these enzymes function as acyl homoserine lactone (AHL) lactonases. Furthermore, we showed that these enzymes inhibited biofilm formation and other quorum-sensing-dependent processes in Pseudomonas aeruginosa, Chromobacterium violaceum, and Agrobacterium tumefaciens. Finally, our experimental data suggest that competitive colonization of roots in the rhizospheres of cowpea plants is affected by DlhR and QsdR1. PMID- 21642402 TI - Electroactivity of phototrophic river biofilms and constitutive cultivable bacteria. AB - Electroactivity is a property of microorganisms assembled in biofilms that has been highlighted in a variety of environments. This characteristic was assessed for phototrophic river biofilms at the community scale and at the bacterial population scale. At the community scale, electroactivity was evaluated on stainless steel and copper alloy coupons used both as biofilm colonization supports and as working electrodes. At the population scale, the ability of environmental bacterial strains to catalyze oxygen reduction was assessed by cyclic voltammetry. Our data demonstrate that phototrophic river biofilm development on the electrodes, measured by dry mass and chlorophyll a content, resulted in significant increases of the recorded potentials, with potentials of up to +120 mV/saturated calomel electrode (SCE) on stainless steel electrodes and +60 mV/SCE on copper electrodes. Thirty-two bacterial strains isolated from natural phototrophic river biofilms were tested by cyclic voltammetry. Twenty five were able to catalyze oxygen reduction, with shifts of potential ranging from 0.06 to 0.23 V, cathodic peak potentials ranging from -0.36 to -0.76 V/SCE, and peak amplitudes ranging from -9.5 to -19.4 MUA. These isolates were diversified phylogenetically (Actinobacteria, Firmicutes, Bacteroidetes, and Alpha-, Beta-, and Gammaproteobacteria) and exhibited various phenotypic properties (Gram stain, oxidase, and catalase characteristics). These data suggest that phototrophic river biofilm communities and/or most of their constitutive bacterial populations present the ability to promote electronic exchange with a metallic electrode, supporting the following possibilities: (i) development of electrochemistry-based sensors allowing in situ phototrophic river biofilm detection and (ii) production of microbial fuel cell inocula under oligotrophic conditions. PMID- 21642403 TI - Determination of microbial diversity of Aeromonas strains on the basis of multilocus sequence typing, phenotype, and presence of putative virulence genes. AB - The genus Aeromonas has been described as comprising several species associated with the aquatic environment, which represents their principal reservoir. Aeromonas spp. are commonly isolated from diseased and healthy fish, but the involvement of such bacteria in human infection and gastroenteritis has frequently been reported. The primary challenge in establishing an unequivocal link between the Aeromonas genus and pathogenesis in humans is the extremely complicated taxonomy. With the aim of clarifying taxonomic relationships among the strains and phenotypes, a multilocus sequencing approach was developed and applied to characterize 23 type and reference strains of Aeromonas spp. and a collection of 77 field strains isolated from fish, crustaceans, and mollusks. All strains were also screened for putative determinants of virulence by PCR (ast, ahh1, act, asa1, eno, ascV, and aexT) and the production of acylated homoserine lactones (AHLs). In addition, the phenotypic fingerprinting obtained from 29 biochemical tests was submitted to the nonparametric combination (NPC) test methodology to define the statistical differences among the identified genetic clusters. Multilocus sequence typing (MLST) achieved precise strain genotyping, and the phylogenetic analysis of concatenated sequences delineated the relationship among the taxa belonging to the genus Aeromonas, providing a powerful tool for outbreak traceability, host range diffusion, and ecological studies. The NPC test showed the feasibility of phenotypic differentiation among the majority of the MLST clusters by using a selection of tests or the entire biochemical fingerprinting. A Web-based MLST sequence database (http://pubmlst.org/aeromonas) specific for the Aeromonas genus was developed and implemented with all the results. PMID- 21642404 TI - Identifying vulnerable pathways in Mycobacterium tuberculosis by using a knockdown approach. AB - We constructed recombinant strains of Mycobacterium tuberculosis in which expression of specific genes was downregulated to identify vulnerable drug targets. Growth phenotypes in macrophages and culture were used to rank targets: the dprE1, clpP1, and fadD32 operons were the best targets and glnA1, glnE, pknL, regX3, and senX3 were poor targets. PMID- 21642405 TI - Identification of loci and functional characterization of trichothecene biosynthesis genes in filamentous fungi of the genus Trichoderma. AB - Trichothecenes are mycotoxins produced by Trichoderma, Fusarium, and at least four other genera in the fungal order Hypocreales. Fusarium has a trichothecene biosynthetic gene (TRI) cluster that encodes transport and regulatory proteins as well as most enzymes required for the formation of the mycotoxins. However, little is known about trichothecene biosynthesis in the other genera. Here, we identify and characterize TRI gene orthologues (tri) in Trichoderma arundinaceum and Trichoderma brevicompactum. Our results indicate that both Trichoderma species have a tri cluster that consists of orthologues of seven genes present in the Fusarium TRI cluster. Organization of genes in the cluster is the same in the two Trichoderma species but differs from the organization in Fusarium. Sequence and functional analysis revealed that the gene (tri5) responsible for the first committed step in trichothecene biosynthesis is located outside the cluster in both Trichoderma species rather than inside the cluster as it is in Fusarium. Heterologous expression analysis revealed that two T. arundinaceum cluster genes (tri4 and tri11) differ in function from their Fusarium orthologues. The Tatri4 encoded enzyme catalyzes only three of the four oxygenation reactions catalyzed by the orthologous enzyme in Fusarium. The Tatri11-encoded enzyme catalyzes a completely different reaction (trichothecene C-4 hydroxylation) than the Fusarium orthologue (trichothecene C-15 hydroxylation). The results of this study indicate that although some characteristics of the tri/TRI cluster have been conserved during evolution of Trichoderma and Fusarium, the cluster has undergone marked changes, including gene loss and/or gain, gene rearrangement, and divergence of gene function. PMID- 21642406 TI - Temporal and spatial variability in culturable pathogenic Vibrio spp. in Lake Pontchartrain, Louisiana, following hurricanes Katrina and Rita. AB - We investigated the abundance, distribution, and virulence gene content of Vibrio cholerae, V. parahaemolyticus, and V. vulnificus in the waters of southern Lake Pontchartrain in Louisiana on four occasions from October 2005 to September 2006, using selective cultivation and molecular assays. The three targeted pathogenic vibrios were generally below the detection level in January 2006, when the water was cold (13 degrees C), and most abundant in September 2006, when the lake water was warmest (30 degrees C). The maximum values for these species were higher than reported previously for the lake by severalfold to orders of magnitude. The only variable consistently correlated with total vibrio abundance within a single sampling was distance from shore (P = 0.000). Multiple linear regression of the entire data set revealed that distance from shore, temperature, and turbidity together explained 82.1% of the variability in total vibrio CFU. The log transformed mean abundance of V. vulnificus CFU in the lake was significantly correlated with temperature (P = 0.014), but not salinity (P = 0.625). Virulence associated genes of V. cholerae (ctx) and V. parahaemolyticus (trh and tdh) were not detected in any isolates of these species (n = 128 and n = 20, respectively). In contrast, 16S rRNA typing of V. vulnificus (n = 298) revealed the presence of both environmental (type A) and clinical (type B) strains. The percentage of the B-type V. vulnificus was significantly higher in the lake in October 2005 (35.8% of the total) than at other sampling times (P <= 0.004), consistent with the view that these strains represent distinct ecotypes. PMID- 21642407 TI - Large-scale comparative phenotypic and genomic analyses reveal ecological preferences of shewanella species and identify metabolic pathways conserved at the genus level. AB - The use of comparative genomics for the study of different microbiological species has increased substantially as sequence technologies become more affordable. However, efforts to fully link a genotype to its phenotype remain limited to the development of one mutant at a time. In this study, we provided a high-throughput alternative to this limiting step by coupling comparative genomics to the use of phenotype arrays for five sequenced Shewanella strains. Positive phenotypes were obtained for 441 nutrients (C, N, P, and S sources), with N-based compounds being the most utilized for all strains. Many genes and pathways predicted by genome analyses were confirmed with the comparative phenotype assay, and three degradation pathways believed to be missing in Shewanella were confirmed as missing. A number of previously unknown gene products were predicted to be parts of pathways or to have a function, expanding the number of gene targets for future genetic analyses. Ecologically, the comparative high-throughput phenotype analysis provided insights into niche specialization among the five different strains. For example, Shewanella amazonensis strain SB2B, isolated from the Amazon River delta, was capable of utilizing 60 C compounds, whereas Shewanella sp. strain W3-18-1, isolated from deep marine sediment, utilized only 25 of them. In spite of the large number of nutrient sources yielding positive results, our study indicated that except for the N sources, they were not sufficiently informative to predict growth phenotypes from increasing evolutionary distances. Our results indicate the importance of phenotypic evaluation for confirming genome predictions. This strategy will accelerate the functional discovery of genes and provide an ecological framework for microbial genome sequencing projects. PMID- 21642408 TI - Temporal variability of coastal Planctomycetes clades at Kabeltonne station, North Sea. AB - Members of the bacterial phylum Planctomycetes are reported in marine water samples worldwide, but quantitative information is scarce. Here we investigated the phylogenetic diversity, abundance, and distribution of Planctomycetes in surface waters off the German North Sea island Helgoland during different seasons by 16S rRNA gene analysis and catalyzed reporter deposition fluorescence in situ hybridization (CARD-FISH). Generally Planctomycetes are more abundant in samples collected in summer and autumn than in samples collected in winter and spring. Statistical analysis revealed that Planctomycetes abundance was correlated to the Centrales diatom bloom in spring 2007. The analysis of size-fractionated seawater samples and of macroaggregates showed that ~90% of the Planctomycetes reside in the >3-MUm size fraction. Comparative sequence analysis of 184 almost full-length 16S rRNA genes revealed three dominant clades. The clades, named Planctomyces related group A, uncultured Planctomycetes group B, and Pirellula-related group D, were monitored by CARD-FISH using newly developed oligonucleotide probes. All three clades showed recurrent abundance patterns during two annual sampling campaigns. Uncultured Planctomycetes group B was most abundant in autumn samples, while Planctomyces-related group A was present in high numbers only during late autumn and winter. The levels of Pirellula-related group D were more constant throughout the year, with elevated counts in summer. Our analyses suggest that the seasonal succession of the Planctomycetes is correlated with algal blooms. We hypothesize that the niche partitioning of the different clades might be caused by their algal substrates. PMID- 21642409 TI - Investigating antibacterial effects of garlic (Allium sativum) concentrate and garlic-derived organosulfur compounds on Campylobacter jejuni by using Fourier transform infrared spectroscopy, Raman spectroscopy, and electron microscopy. AB - Fourier transform infrared (FT-IR) spectroscopy and Raman spectroscopy were used to study the cell injury and inactivation of Campylobacter jejuni from exposure to antioxidants from garlic. C. jejuni was treated with various concentrations of garlic concentrate and garlic-derived organosulfur compounds in growth media and saline at 4, 22, and 35 degrees C. The antimicrobial activities of the diallyl sulfides increased with the number of sulfur atoms (diallyl sulfide < diallyl disulfide < diallyl trisulfide). FT-IR spectroscopy confirmed that organosulfur compounds are responsible for the substantial antimicrobial activity of garlic, much greater than those of garlic phenolic compounds, as indicated by changes in the spectral features of proteins, lipids, and polysaccharides in the bacterial cell membranes. Confocal Raman microscopy (532-nm-gold-particle substrate) and Raman mapping of a single bacterium confirmed the intracellular uptake of sulfur and phenolic components. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were employed to verify cell damage. Principal component analysis (PCA), discriminant function analysis (DFA), and soft independent modeling of class analogs (SIMCA) were performed, and results were cross validated to differentiate bacteria based upon the degree of cell injury. Partial least-squares regression (PLSR) was employed to quantify and predict actual numbers of healthy and injured bacterial cells remaining following treatment. PLSR-based loading plots were investigated to further verify the changes in the cell membrane of C. jejuni treated with organosulfur compounds. We demonstrated that bacterial injury and inactivation could be accurately investigated by complementary infrared and Raman spectroscopies using a chemical based, "whole-organism fingerprint" with the aid of chemometrics and electron microscopy. PMID- 21642410 TI - Differential proteomic analysis of Rickettsia prowazekii propagated in diverse host backgrounds. AB - The obligate intracellular growth of Rickettsia prowazekii places severe restrictions on the analysis of rickettsial gene expression. With a small genome, predicted to code for 835 proteins, identifying which proteins are differentially expressed in rickettsiae that are isolated from different hosts or that vary in virulence is critical to an understanding of rickettsial pathogenicity. We employed a liquid chromatography (LC)-linear trap quadrupole (LTQ)-Orbitrap mass spectrometer for simultaneous acquisition of quantitative mass spectrometry (MS) only data and tandem mass spectrometry (MS-MS) sequence data. With the use of a combination of commercially available algorithms and in-house software, quantitative MS-only data and comprehensive peptide coverage generated from MS-MS were integrated, resulting in the assignment of peptide identities with intensity values, allowing for the differential comparison of complex protein samples. With the use of these protocols, it was possible to directly compare protein abundance and analyze changes in the total proteome profile of R. prowazekii grown in different host backgrounds. Total protein extracted from rickettsiae grown in murine, tick, and insect cell lines or hen egg yolk sacs was analyzed. Here, we report the fold changes, including an upregulation of shock-related proteins, in rickettsiae cultivated in tissue culture compared to the level for rickettsiae harvested from hen yolk sacs. The ability to directly compare, in a complex sample, differential rickettsial protein expression provides a snapshot of host specific proteomic profiles that will help to identify proteins important in intracellular growth and virulence. PMID- 21642411 TI - Sorption and distribution of copper in unsaturated Pseudomonas putida CZ1 biofilms as determined by X-ray fluorescence microscopy. AB - The spatial and temporal distribution of metals in unsaturated Pseudomonas putida CZ1 biofilms was determined using synchrotron-based X-ray fluorescence microscopy (XRF). It was found that Fe, Mn, and Ca were mainly distributed near the air biofilm interface of a biofilm grown on 40 mM citrate, while there were two Fe-, Mn-, and Ca-rich layers within a biofilm grown on 10 mM citrate. The sorption of copper by biofilm grown in medium containing 10 mM citrate was rapid, with copper being found throughout the biofilm after only 1 h of exposure. Copper initially colocalized with Fe and Mn element layers in the biofilm and then precipitated in a 40-MUm-thick layer near the air-biofilm interface when exposed for 12 h. Cu K edge X-ray absorption near edge structure (XANES) analysis revealed that Cu was primarily bound with citrate within the biofilm, and the precipitate formed in the biofilm exposed to copper for 12 h was most similar to copper phosphate. LIVE/DEAD staining revealed that cells at the biofilm-membrane interface were mostly alive even when the copper concentration reached 80.5 mg copper g(-1) biomass. This suggests that the biofilm matrix provided significant protection for cells in this area. These results significantly improve our understanding of metal acquisition, transportation, and immobilization in unsaturated biofilm systems. PMID- 21642412 TI - Discovery of a gene involved in a third bacterial protoporphyrinogen oxidase activity through comparative genomic analysis and functional complementation. AB - Tetrapyrroles are ubiquitous molecules in nearly all living organisms. Heme, an iron-containing tetrapyrrole, is widely distributed in nature, including most characterized aerobic and facultative bacteria. A large majority of bacteria that contain heme possess the ability to synthesize it. Despite this capability and the fact that the biosynthetic pathway has been well studied, enzymes catalyzing at least three steps have remained "missing" in many bacteria. In the current work, we have employed comparative genomics via the SEED genomic platform, coupled with experimental verification utilizing Acinetobacter baylyi ADP1, to identify one of the missing enzymes, a new protoporphyrinogen oxidase, the penultimate enzyme in heme biosynthesis. COG1981 was identified by genomic analysis as a candidate protein family for the missing enzyme in bacteria that lacked HemG or HemY, two known protoporphyrinogen oxidases. The predicted amino acid sequence of COG1981 is unlike those of the known enzymes HemG and HemY, but in some genomes, the gene encoding it is found neighboring other heme biosynthetic genes. When the COG1981 gene was deleted from the genome of A. baylyi, a bacterium that lacks both hemG and hemY, the organism became auxotrophic for heme. Cultures accumulated porphyrin intermediates, and crude cell extracts lacked protoporphyrinogen oxidase activity. The heme auxotrophy was rescued by the presence of a plasmid-borne protoporphyrinogen oxidase gene from a number of different organisms, such as hemG from Escherichia coli, hemY from Myxococcus xanthus, or the human gene for protoporphyrinogen oxidase. PMID- 21642413 TI - Potentially human-pathogenic Escherichia coli O26 in Norwegian sheep flocks. AB - A national survey of Escherichia coli O26 in Norwegian sheep flocks was conducted, using fecal samples to determine the prevalence. In total, 491 flocks were tested, and E. coli O26 was detected in 17.9% of the flocks. One hundred forty-two E. coli O26 isolates were examined for flagellar antigens (H typing) and four virulence genes, including stx and eae, to identify possible Shiga toxin producing E. coli (STEC) and enteropathogenic E. coli (EPEC). Most isolates (129 out of 142) were identified as E. coli O26:H11. They possessed eae and may have potential as human pathogens, although only a small fraction were identified as STEC O26:H11, giving a prevalence in sheep flocks of only 0.8%. Correspondingly, the sheep flock prevalence of atypical EPEC (aEPEC) O26:H11 was surprisingly high (15.9%). The genetic relationship between the E. coli O26:H11 isolates was investigated by pulsed-field gel electrophoresis (PFGE) and multilocus variable number tandem repeat analysis (MLVA), identifying 63 distinct PFGE profiles and 22 MLVA profiles. Although the MLVA protocol was less discriminatory than PFGE and a few cases of disagreement were observed, comparison by partition mapping showed an overall good accordance between the two methods. A close relationship between a few isolates of aEPEC O26:H11 and STEC O26:H11 was identified, but all the E. coli O26:H11 isolates should be considered potentially pathogenic to humans. The present study consisted of a representative sampling of sheep flocks from all parts of Norway. This is the first large survey of sheep flocks focusing on E. coli O26 in general, including results of STEC, aEPEC, and nonpathogenic isolates. PMID- 21642414 TI - Exceptional production of both prodigiosin and cycloprodigiosin as major metabolic constituents by a novel marine bacterium, Zooshikella rubidus S1-1. AB - A Gram-negative, red-pigment-producing marine bacterial strain, designated S1-1, was isolated from the tidal flat sediment of the Yellow Sea, Korea. On the basis of phenotypic, phylogenetic, and genetic data, strain S1-1 (KCTC 11448BP) represented a new species of the genus Zooshikella. Thus, we propose the name Zooshikella rubidus sp. nov. Liquid chromatography and mass spectrometry of the red pigments produced by strain S1-1 revealed that the major metabolic compounds were prodigiosin and cycloprodigiosin. In addition, this organism produced six minor prodigiosin analogues, including two new structures that were previously unknown. To our knowledge, this is the first description of a microorganism that simultaneously produces prodigiosin and cycloprodigiosin as two major metabolites. Both prodigiosin and cycloprodigiosin showed antimicrobial activity against several microbial species. These bacteria were approximately 1.5-fold more sensitive to cycloprodigiosin than to prodigiosin. The metabolites also showed anticancer activity against human melanoma cells, which showed significantly more sensitivity to prodigiosin than to cycloprodigiosin. The secondary metabolite profiles of strain S1-1 and two reference bacterial strains were compared by liquid chromatography-mass spectrometry. Multivariate statistical analyses based on secondary metabolite profiles by liquid chromatography-mass spectrometry indicated that the metabolite profile of strain S1-1 could clearly be distinguished from those of two phylogenetically related, prodigiosin-producing bacterial strains. PMID- 21642415 TI - Redesigning Escherichia coli metabolism for anaerobic production of isobutanol. AB - Fermentation enables the production of reduced metabolites, such as the biofuels ethanol and butanol, from fermentable sugars. This work demonstrates a general approach for designing and constructing a production host that uses a heterologous pathway as an obligately fermentative pathway to produce reduced metabolites, specifically, the biofuel isobutanol. Elementary mode analysis was applied to design an Escherichia coli strain optimized for isobutanol production under strictly anaerobic conditions. The central metabolism of E. coli was decomposed into 38,219 functional, unique, and elementary modes (EMs). The model predictions revealed that during anaerobic growth E. coli cannot produce isobutanol as the sole fermentative product. By deleting 7 chromosomal genes, the total 38,219 EMs were constrained to 12 EMs, 6 of which can produce high yields of isobutanol in a range from 0.29 to 0.41 g isobutanol/g glucose under anaerobic conditions. The remaining 6 EMs rely primarily on the pyruvate dehydrogenase enzyme complex (PDHC) and are typically inhibited under anaerobic conditions. The redesigned E. coli strain was constrained to employ the anaerobic isobutanol pathways through deletion of 7 chromosomal genes, addition of 2 heterologous genes, and overexpression of 5 genes. Here we present the design, construction, and characterization of an isobutanol-producing E. coli strain to illustrate the approach. The model predictions are evaluated in relation to experimental data and strategies proposed to improve anaerobic isobutanol production. We also show that the endogenous alcohol/aldehyde dehydrogenase AdhE is the key enzyme responsible for the production of isobutanol and ethanol under anaerobic conditions. The glycolytic flux can be controlled to regulate the ratio of isobutanol to ethanol production. PMID- 21642416 TI - Synthesis and characterization of chimeric proteins based on cellulase and xylanase from an insect gut bacterium. AB - Insects living on wood and plants harbor a large variety of bacterial flora in their guts for degrading biomass. We isolated a Paenibacillus strain, designated ICGEB2008, from the gut of a cotton bollworm on the basis of its ability to secrete a variety of plant-hydrolyzing enzymes. In this study, we cloned, expressed, and characterized two enzymes, beta-1,4-endoglucanase (Endo5A) and beta-1,4-endoxylanase (Xyl11D), from the ICGEB2008 strain and synthesized recombinant bifunctional enzymes based on Endo5A and Xyl11D. The gene encoding Endo5A was obtained from the genome of the ICGEB2008 strain by shotgun cloning. The gene encoding Xyl11D was obtained using primers for conserved xylanase sequences, which were identified by aligning xylanase sequences in other species of Paenibacillus. Endo5A and Xyl11D were overexpressed in Escherichia coli, and their optimal activities were characterized. Both Endo5A and Xyl11D exhibited maximum specific activity at 50 degrees C and pH 6 to 7. To take advantage of this feature, we constructed four bifunctional chimeric models of Endo5A and Xyl11D by fusing the encoding genes either end to end or through a glycine-serine (GS) linker. We predicted three-dimensional structures of the four models using the I-TASSER server and analyzed their secondary structures using circular dichroism (CD) spectroscopy. The chimeric model Endo5A-GS-Xyl11D, in which a linker separated the two enzymes, yielded the highest C-score on the I-TASSER server, exhibited secondary structure properties closest to the native enzymes, and demonstrated 1.6-fold and 2.3-fold higher enzyme activity than Endo5A and Xyl11D, respectively. This bifunctional enzyme could be effective for hydrolyzing plant biomass owing to its broad substrate range. PMID- 21642417 TI - Prospective blinded comparison of wireless capsule endoscopy and multiphase CT enterography in obscure gastrointestinal bleeding. AB - PURPOSE: To compare the performance of multiphase computed tomographic (CT) enterography with that of capsule endoscopy in a group of patients with obscure gastrointestinal bleeding (OGIB). MATERIALS AND METHODS: This prospective HIPAA compliant study was approved by the institutional review board and the institutional conflict of interest committee. All patients provided written informed consent. Two radiologists, blinded to clinical data and results of capsule endoscopy, interpreted images from CT enterography independently, with discordant interpretations resolved by consensus. Results were compared with those from a reference standard (surgery or endoscopy) and clinical follow-up. Sensitivity and 95% confidence intervals were calculated for each modality. RESULTS: Fifty-eight adult patients, referred for the evaluation of OGIB (occult, 25 patients [43%]; overt, 33 patients [57%]), underwent both tests. A small bowel bleeding source was identified in 16 of the 58 patients (28%). The sensitivity of CT enterography was significantly greater than that of capsule endoscopy (88% [14 of 16 patients] vs 38% [six of 16 patients], respectively; P = .008), largely because it depicted more small bowel masses (100% [nine of nine patients] vs 33% [three of nine patients], respectively; P = .03). No additional small bowel tumors were discovered during the follow-up period (range, 5.6-45.9 months; mean, 16.6 months). CONCLUSION: In this referral population, the sensitivity of CT enterography for detecting small bowel bleeding sources and small bowel masses was significantly greater than that of capsule endoscopy. On the basis of these findings, the addition of multiphase CT enterography to the routine diagnostic work-up of patients with OGIB should be considered, particularly in patients with negative findings at capsule endoscopy. PMID- 21642418 TI - Acute and subacute effects of irreversible electroporation on nerves: experimental study in a pig model. AB - PURPOSE: To evaluate whether irreversible electroporation (IRE) has the potential to damage nerves in a porcine model and to compare histopathologic findings after IRE with histopathologic findings after radiofrequency ablation (RFA). MATERIALS AND METHODS: This study was approved by the institutional animal care and use committee. Computed tomography (CT)-guided IRE of 11 porcine sciatic nerves was performed in nine pigs, and histopathologic analysis was performed on the day of ablation or 3, 6, or 14 days after ablation. In addition, acute RFA of six porcine sciatic nerves was performed in six pigs that were harvested on the day of ablation. All nerves and associated muscles and tissues were assessed for histopathologic findings consistent with athermal or thermal injury, respectively, such as axonal swelling, axonal fragmentation and loss, Wallerian degeneration, inflammatory infiltrates, Schwann cell proliferation, and coagulative necrosis. The percentage of fascicles affected was recorded. RESULTS: All nerves had an axonal injury. The percentage of affected nerve fascicles after IRE was 50%-100%. Axonal swelling and perineural inflammatory infiltrates were detectable at every time point after ablation. Axonal fragmentation and loss, macrophage infiltration, and Schwann cell proliferation were found 6 and 14 days after ablation. Distal Wallerian axonal degeneration was observed 14 days after ablation. The endoneurium and perineurium architecture remained intact in all cases. RFA specimens at the day of ablation revealed acute coagulative necrosis associated with intense basophilic staining of extracellular matrix, including collagen of the perineurium and epineurium consistent with thermal injury. CONCLUSION: IRE has the potential to damage nerves and may result in axonal swelling, fragmentation, and distal Wallerian degeneration. However, preservation of endoneurium architecture and proliferation of Schwann cells may suggest the potential for axonal regeneration. In contrast, RFA leads to thermal nerve damage, causing protein denaturation, and suggests a much lower potential for regeneration. PMID- 21642419 TI - Dry pleural dissemination in non-small cell lung cancer: prognostic and diagnostic implications. AB - PURPOSE: To compare prognostic differences between dry pleural dissemination (DPD) and wet pleural dissemination (WPD) in patients with non-small cell lung cancer (NSCLC) andto review the applicability of computed tomographic (CT) findings of DPD for rendering the diagnosis of this disease. MATERIALS AND METHODS: The institutional review board approved this retrospective study, and informed patient consent was waived. Of 98 patients (male-to-female ratio, 55:43; mean age, 60 years +/- 12) with NSCLC, 20 patients had pathologically proved DPD, and the remaining 78 patients had pathologically proved WPD. Twelve patients, who had been lost to follow-up, were excluded from survival analysis. Observers looked for CT findings of multiple pleural or fissural nodules (more than six in number) and uneven thickening or bandlike thickness. Survival after initial presentation was analyzed and compared between patients with DPD (n = 19) and patients with WPD (n = 67) by using the Kaplan-Meier method and the log-rank test. The sensitivity of CT for depicting DPD was also calculated. RESULTS: Median survival after initial presentation was significantly longer in patients with DPD than in patients with WPD; it was 38 months (95% confidence interval [CI]: 29.9 months, 46.0 months) in patients with DPD and 13 months (95% CI: 9.8 months, 16.2 months) in patients with WPD (P <.001). CT helped identify DPD in 90% (18 of 20) of patients with pathologically proved DPD. Multiple pleural or fissural nodules were noted on CT images in 16 (80%) of 20 patients. Uneven or bandlike pleural thickening was recognized in 15 (75%) patients. CONCLUSION: Patients with DPD show better survival than patients with WPD. CT helps suggest strongly the presence of DPD preoperatively. PMID- 21642420 TI - Organization of the Indian hedgehog--parathyroid hormone-related protein system in the postnatal growth plate. AB - In embryonic growth cartilage, Indian hedgehog (Ihh) and parathyroid hormone related protein (PTHrP) participate in a negative feedback loop that regulates chondrocyte differentiation. Postnatally, this region undergoes major structural and functional changes. To explore the organization of the Ihh-PTHrP system in postnatal growth plate, we microdissected growth plates of 7-day-old rats into their constituent zones and assessed expression of genes participating in the h PTHrP feedback loop. Ihh, Patched 1, Smoothened, Gli1, Gli2, Gli3, and Pthr1 were expressed in regions analogous to the expression domains in embryonic growth cartilage. However, PTHrP was expressed in resting zone cartilage, a site that differs from the embryonic source, the periarticular cells. We then used mice in which lacZ has replaced coding sequences of Gli1 and thus serves as a marker for active hedgehog signaling. At 1, 4, 8, and 12 weeks of age, lacZ expression was detected in a pattern analogous to that of embryonic cartilage. The findings support the hypothesis that the embryonic Ihh-PTHrP feedback loop is maintained in the postnatal growth plate except that the source of PTHrP has shifted to a more proximal location in the resting zone. PMID- 21642421 TI - Intermolecular interactions within the abundant DEAD-box protein Dhh1 regulate its activity in vivo. AB - Dhh1 is a highly conserved DEAD-box protein that has been implicated in many processes involved in mRNA regulation. At least some functions of Dhh1 may be carried out in cytoplasmic foci called processing bodies (P-bodies). Dhh1 was identified initially as a putative RNA helicase based solely on the presence of conserved helicase motifs found in the superfamily 2 (Sf2) of DEXD/H-box proteins. Although initial mutagenesis studies revealed that the signature DEAD box motif is required for Dhh1 function in vivo, enzymatic (ATPase or helicase) or ATP binding activities of Dhh1 or those of any its many higher eukaryotic orthologues have not been described. Here we provide the first characterization of the biochemical activities of Dhh1. Dhh1 has weaker RNA-dependent ATPase activity than other well characterized DEAD-box helicases. We provide evidence that intermolecular interactions between the N- and C-terminal RecA-like helicase domains restrict its ATPase activity; mutation of residues mediating these interactions enhanced ATP hydrolysis. Interestingly, the interdomain interaction mutant displayed enhanced mRNA turnover, RNA binding, and recruitment into cytoplasmic foci in vivo compared with wild type Dhh1. Also, we demonstrate that the ATPase activity of Dhh1 is not required for it to be recruited into cytoplasmic foci, but it regulates its association with RNA in vivo. We hypothesize that the activity of Dhh1 is restricted by interdomain interactions, which can be regulated by cellular factors to impart stringent control over this very abundant RNA helicase. PMID- 21642422 TI - Receptor-independent protein kinase C alpha (PKCalpha) signaling by calpain generated free catalytic domains induces HDAC5 nuclear export and regulates cardiac transcription. AB - Receptor-mediated activation of protein kinase (PK) C is a central pathway regulating cell growth, homeostasis, and programmed death. Recently, we showed that calpain-mediated proteolytic processing of PKCalpha in ischemic myocardium activates PKC signaling in a receptor-independent manner by releasing a persistent and constitutively active free catalytic fragment, PKCalpha-CT. This unregulated kinase provokes cardiomyopathy, but the mechanisms remain unclear. Here, we demonstrate that PKCalpha-CT is a potent regulator of pathological cardiac gene expression. PKCalpha-CT constitutively localizes to nuclei and directly promotes nucleo-cytoplasmic shuttling of HDAC5, inducing expression of apoptosis and other deleterious genes. Whereas PKD activation is required for HDAC5 nuclear export induced by unprocessed PKCs activated by phorbol ester, PKCalpha-CT directly drives HDAC cytosolic relocalization. Activation of MEF2 dependent inflammatory pathway genes by PKCalpha-CT can induce a cell-autonomous transcriptional response that mimics, but anticipates, actual inflammation. Because calpain-mediated processing of PKC isoforms occurs in many tissues wherein calcium is increased by stress or injury, our observation that the catalytically active product of this interaction is a constitutively active transcriptional regulator has broad ramifications for understanding and preventing the pathological transcriptional stress response. PMID- 21642423 TI - Epithelial junctions depend on intercellular trans-interactions between the Na,K ATPase beta1 subunits. AB - N-Glycans of the Na,K-ATPase beta1 subunit are important for intercellular adhesion in epithelia, suggesting that epithelial junctions depend on N-glycan mediated interactions between the beta1 subunits of neighboring cells. The level of co-immunoprecipitation of the endogenous beta1 subunit with various YFP-linked beta1 subunits expressed in Madin-Darby canine kidney cells was used to assess beta1-beta1 interactions. The amount of co-precipitated endogenous dog beta1 was greater with dog YFP-beta1 than with rat YFP-beta1, showing that amino acid mediated interactions are important for beta1-beta1 binding. Co-precipitation of beta1 was also less with the unglycosylated YFP-beta1 than with glycosylated YFP beta1, indicating a role for N-glycans. Mixing cells expressing dog YFP-beta1 with non-transfected cells increased the amount of co-precipitated beta1, confirming the presence of intercellular (YFP-beta1)-beta1 complexes. Accordingly, disruption of intercellular junctions decreased the amount of co precipitated beta1 subunits. The decrease in beta1 co-precipitation both with rat YFP-beta1 and unglycosylated YFP-beta1 was associated with decreased detergent stability of junctional proteins and increased paracellular permeability. Reducing N-glycan branching by specific inhibitors increased (YFP-beta1)-beta1 co precipitation and strengthened intercellular junctions. Therefore, interactions between the beta1 subunits of neighboring cells maintain integrity of intercellular junctions, and alterations in the beta1 subunit N-glycan structure can regulate stability and tightness of intercellular junctions. PMID- 21642424 TI - Loss of cleavage at beta'-site contributes to apparent increase in beta-amyloid peptide (Abeta) secretion by beta-secretase (BACE1)-glycosylphosphatidylinositol (GPI) processing of amyloid precursor protein. AB - Several lines of evidence implicate lipid raft microdomains in Alzheimer disease associated beta-amyloid peptide (Abeta) production. Notably, targeting beta secretase (beta-site amyloid precursor protein (APP)-cleaving enzyme 1 (BACE1)) exclusively to lipid rafts by the addition of a glycosylphosphatidylinositol (GPI) anchor to its ectodomain has been reported to elevate Abeta secretion. Paradoxically, Abeta secretion is not reduced by the expression of non-raft resident S-palmitoylation-deficient BACE1 (BACE1-4C/A (C474A/C478A/C482A/C485A)). We addressed this apparent discrepancy in raft microdomain-associated BACE1 processing of APP in this study. As previously reported, we found that expression of BACE1-GPI elevated Abeta secretion as compared with wild-type BACE1 (WTBACE1) or BACE1-4C/A. However, this increase occurred without any difference in the levels of APP ectodomain released following BACE1 cleavage (soluble APPbeta), arguing against an overall increase in BACE1 processing of APP per se. Further analysis revealed that WTBACE1 cleaves APP at beta- and beta'-sites, generating +1 and +11 beta-C-terminal fragments and secreting intact as well as N-terminally truncated Abeta. In contrast, three different BACE1-GPI chimeras preferentially cleaved APP at the beta-site, mainly generating +1 beta-C-terminal fragment and secreting intact Abeta. As a consequence, cells expressing BACE1-GPI secreted relatively higher levels of intact Abeta without an increase in BACE1 processing of APP. Markedly reduced cleavage at beta'-site exhibited by BACE1-GPI was cell type-independent and insensitive to subcellular localization of APP or the pathogenic KM/NL mutant. We conclude that the apparent elevation in Abeta secretion by BACE1-GPI is mainly attributed to preferential cleavage at the beta site and failure to detect +11 Abeta species secreted by cells expressing WTBACE1. PMID- 21642425 TI - Unsaturated fatty acids drive disintegrin and metalloproteinase (ADAM)-dependent cell adhesion, proliferation, and migration by modulating membrane fluidity. AB - The disintegrin-metalloproteinases ADAM10 and ADAM17 mediate the release of several cell signaling molecules and cell adhesion molecules such as vascular endothelial cadherin or L-selectin affecting endothelial permeability and leukocyte transmigration. Dysregulation of ADAM activity may contribute to the pathogenesis of vascular diseases, but the mechanisms underlying the control of ADAM functions are still incompletely understood. Atherosclerosis is characterized by lipid plaque formation and local accumulation of unsaturated free fatty acids (FFA). Here, we show that unsaturated FFA increase ADAM-mediated substrate cleavage. We demonstrate that these alterations are not due to genuine changes in enzyme activity, but correlate with changes in membrane fluidity as revealed by measurement of 1,6-diphenyl-1,3,5-hexatriene fluorescence anisotropy and fluorescence recovery after photobleaching analyses. ELISA and immunoblot experiments conducted with granulocytes, endothelial cells, and keratinocytes revealed rapid increase of ectodomain shedding of ADAM10 and ADAM17 substrates upon membrane fluidization. Large amounts of unsaturated FFA may be liberated from cholesteryl esters in LDL that is entrapped in atherosclerotic lesions. Incubation of cells with thus modified LDL resulted in rapid cleavage of ADAM substrates with corresponding functional consequences on cell proliferation, cell migration, and endothelial permeability, events of high significance in atherogenesis. We propose that FFA represent critical regulators of ADAM function that may assume relevance in many biological settings through their influence on mobility of enzyme and substrate in lipid bilayers. PMID- 21642426 TI - Allosteric communication between the nucleotide binding domains of caseinolytic peptidase B. AB - ClpB is a hexameric chaperone that solubilizes and reactivates protein aggregates in cooperation with the Hsp70/DnaK chaperone system. Each of the identical protein monomers contains two nucleotide binding domains (NBD), whose ATPase activity must be coupled to exert on the substrate the mechanical work required for its reactivation. However, how communication between these sites occurs is at present poorly understood. We have studied herein the affinity of each of the NBDs for nucleotides in WT ClpB and protein variants in which one or both sites are mutated to selectively impair nucleotide binding or hydrolysis. Our data show that the affinity of NBD2 for nucleotides (K(d) = 3-7 MUm) is significantly higher than that of NBD1. Interestingly, the affinity of NBD1 depends on nucleotide binding to NBD2. Binding of ATP, but not ADP, to NBD2 increases the affinity of NBD1 (the K(d) decreases from ~160-300 to 50-60 MUm) for the corresponding nucleotide. Moreover, filling of the NBD2 ring with ATP allows the cooperative binding of this nucleotide and substrates to the NBD1 ring. Data also suggest that a minimum of four subunits cooperate to bind and reactivate two different aggregated protein substrates. PMID- 21642427 TI - The ETS family transcription factor ELK-1 regulates induction of the cell cycle regulatory gene p21(Waf1/Cip1) and the BAX gene in sodium arsenite-exposed human keratinocyte HaCaT cells. AB - Cyclin-dependent kinase inhibitor (CDKN1A), often referred to as p21(Waf1/Cip1) (p21), is induced by a variety of environmental stresses. Transcription factor ELK-1 is a member of the ETS oncogene superfamily. Here, we show that ELK-1 directly trans-activates the p21 gene, independently of p53 and EGR-1, in sodium arsenite (NaASO(2))-exposed HaCaT cells. Promoter deletion analysis and site directed mutagenesis identified the presence of an ELK-1-binding core motif between -190 and -170 bp of the p21 promoter that confers inducibility by NaASO(2). Chromatin immunoprecipitation and electrophoretic mobility shift analyses confirmed the specific binding of ELK-1 to its putative binding sequence within the p21 promoter. In addition, NaASO(2)-induced p21 promoter activity was enhanced by exogenous expression of ELK-1 and reduced by expression of siRNA targeted to ELK-1 mRNA. The importance of ELK-1 in response to NaASO(2) was further confirmed by the observation that stable expression of ELK-1 siRNA in HaCaT cells resulted in the attenuation of NaASO(2)-induced p21 expression. Although ELK-1 was activated by ERK, JNK, and p38 MAPK in response to NaASO(2), ELK-1-mediated activation of the p21 promoter was largely dependent on ERK. In addition, EGR-1 induced by ELK-1 seemed to be involved in NaASO(2)-induced expression of BAX. This supports the view that the ERK/ELK-1 cascade is involved in p53-independent induction of p21 and BAX gene expression. PMID- 21642428 TI - Permeabilization of the mitochondrial outer membrane by Bax/truncated Bid (tBid) proteins as sensitized by cardiolipin hydroperoxide translocation: mechanistic implications for the intrinsic pathway of oxidative apoptosis. AB - Cytochrome c (cyt c) release upon oxidation of cardiolipin (CL) in the mitochondrial inner membrane (IM) under oxidative stress occurs early in the intrinsic apoptotic pathway. We postulated that CL oxidation mobilizes not only cyt c but also CL itself in the form of hydroperoxide (CLOOH) species. Relatively hydrophilic CLOOHs could assist in apoptotic signaling by translocating to the outer membrane (OM), thus promoting recruitment of the pro-apoptotic proteins truncated Bid (tBid) and Bax for generation of cyt c-traversable pores. Initial testing of these possibilities showed that CLOOH-containing liposomes were permeabilized more readily by tBid plus Ca(2+) than CL-containing counterparts. Moreover, CLOOH translocated more rapidly from IM-mimetic to OM-mimetic liposomes than CL and permitted more extensive OM permeabilization. We found that tBid bound more avidly to CLOOH-containing membranes than to CL counterparts, and binding increased with increasing CLOOH content. Permeabilization of CLOOH containing liposomes in the presence of tBid could be triggered by monomeric Bax, consistent with tBid/Bax cooperation in pore formation. Using CL-null mitochondria from a yeast mutant, we found that tBid binding and cyt c release were dramatically enhanced by transfer acquisition of CLOOH. Additionally, we observed a pre-apoptotic IM-to-OM transfer of oxidized CL in cardiomyocytes treated with the Complex III blocker, antimycin A. These findings provide new mechanistic insights into the role of CL oxidation in the intrinsic pathway of oxidative apoptosis. PMID- 21642429 TI - Cu(II) mediates kinetically distinct, non-amyloidogenic aggregation of amyloid beta peptides. AB - Cu(II) ions are implicated in the pathogenesis of Alzheimer disease by influencing the aggregation of the amyloid-beta (Abeta) peptide. Elucidating the underlying Cu(II)-induced Abeta aggregation is paramount for understanding the role of Cu(II) in the pathology of Alzheimer disease. The aim of this study was to characterize the qualitative and quantitative influence of Cu(II) on the extracellular aggregation mechanism and aggregate morphology of Abeta(1-40) using spectroscopic, microelectrophoretic, mass spectrometric, and ultrastructural techniques. We found that the Cu(II):Abeta ratio in solution has a major influence on (i) the aggregation kinetics/mechanism of Abeta, because three different kinetic scenarios were observed depending on the Cu(II):Abeta ratio, (ii) the metal:peptide stoichiometry in the aggregates, which increased to 1.4 at supra-equimolar Cu(II):Abeta ratio; and (iii) the morphology of the aggregates, which shifted from fibrillar to non-fibrillar at increasing Cu(II):Abeta ratios. We observed dynamic morphological changes of the aggregates, and that the formation of spherical aggregates appeared to be a common morphological end point independent on the Cu(II) concentration. Experiments with Abeta(1-42) were compatible with the conclusions for Abeta(1-40) even though the low solubility of Abeta(1-42) precluded examination under the same conditions as for the Abeta(1 40). Experiments with Abeta(1-16) and Abeta(1-28) showed that other parts than the Cu(II)-binding His residues were important for Cu(II)-induced Abeta aggregation. Based on this study we propose three mechanistic models for the Cu(II)-induced aggregation of Abeta(1-40) depending on the Cu(II):Abeta ratio, and identify key reaction steps that may be feasible targets for preventing Cu(II)-associated aggregation or toxicity in Alzheimer disease. PMID- 21642430 TI - Biochemical mapping of interactions within the intraflagellar transport (IFT) B core complex: IFT52 binds directly to four other IFT-B subunits. AB - Cilia and flagella are complex structures emanating from the surface of most eukaroytic cells and serve important functions including motility, signaling, and sensory reception. A process called intraflagellar transport (IFT) is of central importance to ciliary assembly and maintenance. The IFT complex is required for this transport and consists of two distinct multisubunit subcomplexes, IFT-A and IFT-B. Despite the importance of the IFT complex, little is known about its overall architecture. This paper presents a biochemical dissection of the molecular interactions within the IFT-B core complex. Two stable subcomplexes consisting of IFT88/70/52/46 and IFT81/74/27/25 were recombinantly co-expressed and purified. We identify a novel interaction between IFT70/52 and map the interaction domains between IFT52 and the other subunits within the IFT88/70/52/46 complex. Additionally, we show that IFT52 binds directly to the IFT81/74/27/25 complex, indicating that it could mediate the interaction between the two subcomplexes. Our data lead to an improved architectural map for the IFT B core complex with new interactions as well as domain resolution mapping for several subunits. PMID- 21642431 TI - New family of deamination repair enzymes in uracil-DNA glycosylase superfamily. AB - DNA glycosylases play a major role in the repair of deaminated DNA damage. Previous investigations identified five families within the uracil-DNA glycosylase (UDG) superfamily. All enzymes within the superfamily studied thus far exhibit uracil-DNA glycosylase activity. Here we identify a new class of DNA glycosylases in the UDG superfamily that lacks UDG activity. Instead, these enzymes act as hypoxanthine-DNA glycosylases in vitro and in vivo. Molecular modeling and structure-guided mutational analysis allowed us to identify a unique catalytic center in this class of DNA glycosylases. Based on unprecedented biochemical properties and phylogenetic analysis, we propose this new class of DNA repair glycosylases that exists in bacteria, archaea, and eukaryotes as family 6 and designate it as the hypoxanthine-DNA glycosylase family. This study demonstrates the structural evolvability that underlies substrate specificity and catalytic flexibility in the evolution of enzymatic function. PMID- 21642432 TI - Rtt107 is required for recruitment of the SMC5/6 complex to DNA double strand breaks. AB - Genome integrity is maintained by a network of DNA damage response pathways, including checkpoints and DNA repair processes. In Saccharomyces cerevisiae, the BRCT domain-containing protein Rtt107/Esc4 is required for the restart of DNA replication after successful repair of DNA damage and for cellular resistance to DNA-damaging agents. In addition to its well characterized interaction with the endonuclease Slx4, Rtt107 interacts with a number of other DNA repair and recombination proteins. These include the evolutionarily conserved SMC5/6 complex, which is involved in numerous chromosome maintenance activities, such as DNA repair, chromosome segregation, and telomere function. The interaction between Rtt107 and the SMC5/6 complex was mediated through the N-terminal BRCT domains of Rtt107 and the Nse6 subunit of SMC5/6 and was independent of methyl methane sulfonate-induced damage and Slx4. Supporting a shared function in the DNA damage response, Rtt107 was required for recruitment of SMC5/6 to DNA double strand breaks. However, this functional relationship did not extend to other types of DNA lesions such as protein-bound nicks. Interestingly, Rtt107 was phosphorylated when SMC5/6 function was compromised in the absence of DNA damaging agents, indicating a connection beyond the DNA damage response. Genetic analyses revealed that, although a subset of Rtt107 and SMC5/6 functions was shared, these proteins also contributed independently to maintenance of genome integrity. PMID- 21642433 TI - Heparin impairs angiogenesis through inhibition of microRNA-10b. AB - Heparin, which has been used as an anticoagulant drug for decades, inhibits angiogenesis, whereas thrombin promotes tumor-associated angiogenesis. However, the mechanisms underlying the regulation of angiogenesis by heparin and thrombin are not well understood. Here, we show that microRNA-10b (miR-10b) is down regulated by heparin and up-regulated by thrombin in human microvascular endothelial cells (HMEC-1). Overexpression of miR-10b induces HMEC-1 cell migration, tube formation, and angiogenesis, and down-regulates homeobox D10 (HoxD10) expression via direct binding of miR-10b to the putative 3' UTR of HoxD10. In addition, HMEC-1 cell migration and tube formation are induced by HoxD10 knockdown, whereas angiogenesis is arrested when HoxD10 expression is increased after anti-miR-10b or heparin treatments. Furthermore, expression of miR-10b and its transcription factor Twist are up-regulated by thrombin, whereas HoxD10 expression is impaired by thrombin. Using quartz crystal microbalance analysis, we show that heparin binds to thrombin, thereby inhibiting thrombin induced expression of Twist and miR-10b. However, the expression of miR-10b is not attenuated by heparin any more after thrombin expression is silenced by its siRNA. Interestingly, we find that heparin attenuates miR-10b expression and induces HoxD10 expression in vivo to inhibit angiogenesis and impair the growth of MDA-MB-231 tumor xenografts. These results provide insight into the molecular mechanism by which heparin and thrombin regulate angiogenesis. PMID- 21642434 TI - Type XVII collagen regulates lamellipod stability, cell motility, and signaling to Rac1 by targeting bullous pemphigoid antigen 1e to alpha6beta4 integrin. AB - Rac1 activity, polarity, lamellipodial dynamics, and directed motility are defective in keratinocytes exhibiting deficiency in beta4 integrin or knockdown of the plakin protein Bullous Pemphigoid Antigen 1e (BPAG1e). The activity of Rac, formation of stable lamellipodia, and directed migration are restored in beta4 integrin-deficient cells by inducing expression of a truncated form of beta4 integrin, which lacks binding sites for BPAG1e and plectin. In these same cells, BPAG1e, the truncated beta4 integrin, and type XVII collagen (Col XVII), a transmembrane BPAG1e-binding protein, but not plectin, colocalize along the substratum-attached surface. This finding suggested to us that Col XVII mediates the association of BPAG1e and alpha6beta4 integrin containing the truncated beta4 subunit and supports directed migration. To test these possibilities, we knocked down Col XVII expression in keratinocytes expressing both full-length and truncated beta4 integrin proteins. Col XVII-knockdown keratinocytes exhibit a loss in BPAG1e-alpha6beta4 integrin interaction, a reduction in lamellipodial stability, an impairment in directional motility, and a decrease in Rac1 activity. These defects are rescued by a mutant Col XVII protein truncated at its carboxyl terminus. In summary, our results suggest that in motile cells Col XVII recruits BPAG1e to alpha6beta4 integrin and is necessary for activation of signaling pathways, motile behavior, and lamellipodial stability. PMID- 21642435 TI - Suppression of amyloid beta A11 antibody immunoreactivity by vitamin C: possible role of heparan sulfate oligosaccharides derived from glypican-1 by ascorbate induced, nitric oxide (NO)-catalyzed degradation. AB - Amyloid beta (Abeta) is generated from the copper- and heparan sulfate (HS) binding amyloid precursor protein (APP) by proteolytic processing. APP supports S nitrosylation of the HS proteoglycan glypican-1 (Gpc-1). In the presence of ascorbate, there is NO-catalyzed release of anhydromannose (anMan)-containing oligosaccharides from Gpc-1-nitrosothiol. We investigated whether these oligosaccharides interact with Abeta during APP processing and plaque formation. anMan immunoreactivity was detected in amyloid plaques of Alzheimer (AD) and APP transgenic (Tg2576) mouse brains by immunofluorescence microscopy. APP/APP degradation products detected by antibodies to the C terminus of APP, but not Abeta oligomers detected by the anti-Abeta A11 antibody, colocalized with anMan immunoreactivity in Tg2576 fibroblasts. A 50-55-kDa anionic, sodium dodecyl sulfate-stable, anMan- and Abeta-immunoreactive species was obtained from Tg2576 fibroblasts using immunoprecipitation with anti-APP (C terminus). anMan containing HS oligo- and disaccharide preparations modulated or suppressed A11 immunoreactivity and oligomerization of Abeta42 peptide in an in vitro assay. A11 immunoreactivity increased in Tg2576 fibroblasts when Gpc-1 autoprocessing was inhibited by 3-beta[2(diethylamino)ethoxy]androst-5-en-17-one (U18666A) and decreased when Gpc-1 autoprocessing was stimulated by ascorbate. Neither overexpression of Gpc-1 in Tg2576 fibroblasts nor addition of copper ion and NO donor to hippocampal slices from 3xTg-AD mice affected A11 immunoreactivity levels. However, A11 immunoreactivity was greatly suppressed by the subsequent addition of ascorbate. We speculate that temporary interaction between the Abeta domain and small, anMan-containing oligosaccharides may preclude formation of toxic Abeta oligomers. A portion of the oligosaccharides are co-secreted with the Abeta peptides and deposited in plaques. These results support the notion that an inadequate supply of vitamin C could contribute to late onset AD in humans. PMID- 21642436 TI - Oxidation of 3,4-dihydroxyphenylacetaldehyde, a toxic dopaminergic metabolite, to a semiquinone radical and an ortho-quinone. AB - The oxidation and toxicity of dopamine is believed to contribute to the selective neurodegeneration associated with Parkinson disease. The formation of reactive radicals and quinones greatly contributes to dopaminergic toxicity through a variety of mechanisms. The physiological metabolism of dopamine to 3,4 dihydroxyphenylacetaldehyde (DOPAL) via monoamine oxidase significantly increases its toxicity. To more adequately explain this enhanced toxicity, we hypothesized that DOPAL is capable of forming radical and quinone species upon oxidation. Here, two unique oxidation products of DOPAL are identified. Several different oxidation methods gave rise to a transient DOPAL semiquinone radical, which was characterized by electron paramagnetic resonance spectroscopy. NMR identified the second oxidation product of DOPAL as the ortho-quinone. Also, carbonyl hydration of DOPAL in aqueous media was evident via NMR. Interestingly, the DOPAL quinone exists exclusively in the hydrated form. Furthermore, the enzymatic and chemical oxidation of DOPAL greatly enhance protein cross-linking, whereas auto-oxidation results in the production of superoxide. Also, DOPAL was shown to be susceptible to oxidation by cyclooxygenase-2 (COX-2). The involvement of this physiologically relevant enzyme in both oxidative stress and Parkinson disease underscores the potential importance of DOPAL in the pathogenesis of this condition. PMID- 21642437 TI - Dentin matrix protein 1 (DMP1) signals via cell surface integrin. AB - Dentin matrix phosphoprotein 1 (DMP1) is a non-collagenous, acidic extracellular matrix protein expressed chiefly in bone and dentin. We examined the DMP1 ability to engage cell-surface receptors and subsequently activate intracellular signaling pathways. Our data indeed show that the presence of extracellular DMP1 triggers focal adhesion point formation in human mesenchymal stem cells and osteoblast-like cells. We determine that DMP1 acts via interaction with alphavbeta3 integrin and stimulates phosphorylation of focal adhesion kinase. Further biochemical characterization confirms the activation of downstream effectors of the MAPK pathways, namely ERK and JNK, after DMP1 treatment. This activation is specifically inhibitable and can also be blocked by the addition of anti-alphavbeta3 integrin antibody. Furthermore, we show that extracellular treatment with DMP1 stimulates the translocation of phosphorylated JNK to the nucleus and a concomitant up-regulation of transcriptional activation by phosphorylated c-Jun. The evidence presented here indicates that DMP1 is specifically involved in signaling via extracellular matrix-cell surface interaction. Combined with the published DMP1-null data (Feng, J. Q., Ward, L. M., Liu, S., Lu, Y., Xie, Y., Yuan, B., Yu, X., Rauch, F., Davis, S. I., Zhang, S., Rios, H., Drezner, M. K., Quarles, L. D., Bonewald, L. F., and White, K. E. (2006) Nat. Genet. 38, 1310-1315) it can be hypothesized that DMP1 could be a key effector of ECM-osteocyte signaling. PMID- 21642438 TI - Fibrinogen is a ligand for the Staphylococcus aureus microbial surface components recognizing adhesive matrix molecules (MSCRAMM) bone sialoprotein-binding protein (Bbp). AB - Microbial surface components recognizing adhesive matrix molecules (MSCRAMMs) are bacterial surface proteins mediating adherence of the microbes to components of the extracellular matrix of the host. On Staphylococci, the MSCRAMMs often have multiple ligands. Consequently, we hypothesized that the Staphylococcus aureus MSCRAMM bone sialoprotein-binding protein (Bbp) might recognize host molecules other than the identified bone protein. A ligand screen revealed that Bbp binds human fibrinogen (Fg) but not Fg from other mammals. We have characterized the interaction between Bbp and Fg. The binding site for Bbp was mapped to residues 561-575 in the Fg Aalpha chain using recombinant Fg chains and truncation mutants in Far Western blots and solid-phase binding assays. Surface plasmon resonance was used to determine the affinity of Bbp for Fg. The interaction of Bbp with Fg peptides corresponding to the mapped residues was further characterized using isothermal titration calorimetry. In addition, Bbp expressed on the surface of bacteria mediated adherence to immobilized Fg Aalpha. Also, Bbp interferes with thrombin-induced Fg coagulation. Together these data demonstrate that human Fg is a ligand for Bbp and that Bbp can manipulate the biology of the Fg ligand in the host. PMID- 21642439 TI - The multidrug efflux pump MdtEF protects against nitrosative damage during the anaerobic respiration in Escherichia coli. AB - Drug efflux represents an important protection mechanism in bacteria to withstand antibiotics and environmental toxic substances. Efflux genes constitute 6-18% of all transporters in bacterial genomes, yet the expression and functions of only a handful of them have been studied. Among the 20 efflux genes encoded in the Escherichia coli K-12 genome, only the AcrAB-TolC system is constitutively expressed. The expression, activities, and physiological functions of the remaining efflux genes are poorly understood. In this study we identified a dramatic up-regulation of an additional efflux pump, MdtEF, under the anaerobic growth condition of E. coli, which is independent of antibiotic exposure. We found that expression of MdtEF is up-regulated more than 20-fold under anaerobic conditions by the global transcription factor ArcA, resulting in increased efflux activity and enhanced drug tolerance in anaerobically grown E. coli. Cells lacking mdtEF display a significantly decreased survival rate under the condition of anaerobic respiration of nitrate. Deletion of the genes responsible for the biosynthesis of indole, tnaAB, or replacing nitrate with fumarate as the terminal electron acceptor during the anaerobic respiration restores the decreased survival of DeltamdtEF cells. Moreover, DeltamdtEF cells are susceptible to indole nitrosative derivatives, a class of toxic byproducts formed and accumulated within E. coli when the bacterium respires nitrate under anaerobic conditions. Taken together, we conclude that the multidrug efflux pump MdtEF is up-regulated during the anaerobic physiology of E. coli to protect the bacterium from nitrosative damage through expelling the nitrosyl indole derivatives out of the cells. PMID- 21642440 TI - Actin filament bundling by fimbrin is important for endocytosis, cytokinesis, and polarization in fission yeast. AB - Through the coordinated action of diverse actin-binding proteins, cells simultaneously assemble actin filaments with distinct architectures and dynamics to drive different processes. Actin filament cross-linking proteins organize filaments into higher order networks, although the requirement of cross-linking activity in cells has largely been assumed rather than directly tested. Fission yeast Schizosaccharomyces pombe assembles actin into three discrete structures: endocytic actin patches, polarizing actin cables, and the cytokinetic contractile ring. The fission yeast filament cross-linker fimbrin Fim1 primarily localizes to Arp2/3 complex-nucleated branched filaments of the actin patch and by a lesser amount to bundles of linear antiparallel filaments in the contractile ring. It is unclear whether Fim1 associates with bundles of parallel filaments in actin cables. We previously discovered that a principal role of Fim1 is to control localization of tropomyosin Cdc8, thereby facilitating cofilin-mediated filament turnover. Therefore, we hypothesized that the bundling ability of Fim1 is dispensable for actin patches but is important for the contractile ring and possibly actin cables. By directly visualizing actin filament assembly using total internal reflection fluorescence microscopy, we determined that Fim1 bundles filaments in both parallel and antiparallel orientations and efficiently bundles Arp2/3 complex-branched filaments in the absence but not the presence of actin capping protein. Examination of cells exclusively expressing a truncated version of Fim1 that can bind but not bundle actin filaments revealed that bundling activity of Fim1 is in fact important for all three actin structures. Therefore, fimbrin Fim1 has diverse roles as both a filament "gatekeeper" and as a filament cross-linker. PMID- 21642441 TI - Retinoic acid-inducible gene I-inducible miR-23b inhibits infections by minor group rhinoviruses through down-regulation of the very low density lipoprotein receptor. AB - In mammals, viral infections are detected by innate immune receptors, including Toll-like receptor and retinoic acid inducible gene I (RIG-I)-like receptor (RLR), which activate the type I interferon (IFN) system. IFN essentially activates genes encoding antiviral proteins that inhibit various steps of viral replication as well as facilitate the subsequent activation of acquired immune responses. In this study, we investigated the expression of non-coding RNA upon viral infection or RLR activation. Using a microarray, we identified several microRNAs (miRNA) specifically induced to express by RLR signaling. As suggested by Bioinformatics (miRBase Target Data base), one of the RLR-inducible miRNAs, miR-23b, actually knocked down the expression of very low density lipoprotein receptor (VLDLR) and LDLR-related protein 5 (LRP5). Transfection of miR-23b specifically inhibited infection of rhinovirus 1B (RV1B), which utilizes the low density lipoprotein receptor (LDLR) family for viral entry. Conversely, introduction of anti-miRNA-23b enhanced the viral yield. Knockdown experiments using small interfering RNA (siRNA) revealed that VLDLR, but not LRP5, is critical for an efficient infection by RV1B. Furthermore, experiments with the transfection of infectious viral RNA revealed that miR-23b did not affect post entry viral replication. Our results strongly suggest that RIG-I signaling results in the inhibitions of infections of RV1B through the miR-23b-mediated down-regulation of its receptor VLDLR. PMID- 21642443 TI - Random variation and rankability of hospitals using outcome indicators. AB - OBJECTIVE: There is a growing focus on quality and safety in healthcare. Outcome indicators are increasingly used to compare hospital performance and to rank hospitals, but the reliability of ranking (rankability) is under debate. This study aims to quantify the rankability of several outcome indicators of hospital performance currently used by the Dutch government. METHODS: From 52 indicators used by the Netherlands Inspectorate, the authors selected nine outcome indicators presenting a fraction and absolute numbers. Of these indicators, four were combined into two, resulting in seven indicators for analysis. The official data of 97 Dutch hospitals for the year 2007 were used. Uncertainty in the observed outcomes within the hospitals (within hospital variance, sigma(2)) was estimated using fixed effect logistic regression models. Heterogeneity (between hospital variance, tau(2)) was measured with random effect logistic regression models. Subsequently, the rankability was calculated by relating heterogeneity to uncertainty within and between hospitals (tau(2)/(tau(2) +median sigma(2))). RESULTS: Sample sizes varied but were typically around 200 per hospital (range of median 90-277) with a median of 2-21 cases, causing a substantial uncertainty in outcomes per hospital. Although fourfold to eightfold differences between hospitals were noted, the uncertainty within hospitals caused a poor (<50%) rankability in three indicators and moderate rankability (50-75%) in the other four indicators. CONCLUSION: The currently used Dutch outcome indicators are not suitable for ranking hospitals. When judging hospital quality the influence of random variation must be accounted for to avoid overinterpretation of the numbers in the quest for more transparency in healthcare. Adequate sample size is a prerequisite in attempting reliable ranking. PMID- 21642442 TI - Evolution of the PEBP gene family in plants: functional diversification in seed plant evolution. AB - The phosphatidyl ethanolamine-binding protein (PEBP) gene family is present in all eukaryote kingdoms, with three subfamilies identified in angiosperms (FLOWERING LOCUS T [FT], MOTHER OF FT AND TFL1 [MFT], and TERMINAL FLOWER1 [TFL1] like). In angiosperms, PEBP genes have been shown to function both as promoters and suppressors of flowering and to control plant architecture. In this study, we focus on previously uncharacterized PEBP genes from gymnosperms. Extensive database searches suggest that gymnosperms possess only two types of PEBP genes, MFT-like and a group that occupies an intermediate phylogenetic position between the FT-like and TFL1-like (FT/TFL1-like). Overexpression of Picea abies PEBP genes in Arabidopsis (Arabidopsis thaliana) suggests that the FT/TFL1-like genes (PaFTL1 and PaFTL2) code for proteins with a TFL1-like function. However, PaFTL1 and PaFTL2 also show highly divergent expression patterns. While the expression of PaFTL2 is correlated with annual growth rhythm and mainly confined to needles and vegetative and reproductive buds, the expression of PaFTL1 is largely restricted to microsporophylls of male cones. The P. abies MFT-like genes (PaMFT1 and PaMFT2) show a predominant expression during embryo development, a pattern that is also found for many MFT-like genes from angiosperms. P. abies PEBP gene expression is primarily detected in tissues undergoing physiological changes related to growth arrest and dormancy. A first duplication event resulting in two families of plant PEBP genes (MFT-like and FT/TFL1-like) seems to coincide with the evolution of seed plants, in which independent control of bud and seed dormancy was required, and the second duplication resulting in the FT-like and TFL1-like clades probably coincided with the evolution of angiosperms. PMID- 21642444 TI - Healthcare-associated infections must stop: a breakthrough project aimed at reducing healthcare-associated infections in an intensive-care unit. AB - BACKGROUND: Healthcare-associated infections are a problem for the Swedish healthcare system. In order to reduce the risk of acquiring healthcare-associated infections, the intensive-care unit attended a breakthrough project in 2004-2005, with the aims of studying methods of increasing patient safety and systematically improving treatment outcomes. The intensive-care unit had no system for registering infections, and the authors wanted to ascertain the prevalence of healthcare-associated infections, and register and prevent them. OBJECTIVES: 40% reduction in healthcare-associated infections in ventilated patients. 100% of staff to implement basic hygiene routines. DESIGN: The method used was the Breakthrough Series, originally designed by the Institute for Healthcare Improvement in Boston, Massachusetts, USA. The method aims to bridge the gap between what is known and what is done, spreading best-practice methods even faster. Many ideas for changes are tested on a small scale, with the basic rule that the small changes combine to create large changes that lead towards the final goal. RESULTS: The frequency of healthcare-associated infections in ventilated patients was reduced by 43%. Compliance with the basic hygiene routines improved greatly, from 72% on initial measurement to 98% today. CONCLUSION: In order to reduce the risk of acquiring healthcare-associated infections and to increase patient safety, a continuous, systematic effort involving continual measurement and review is necessary. PMID- 21642445 TI - Assessment of potential toxicity of a smokeless tobacco product (naswar) available on the Pakistani market. AB - BACKGROUND: 'Naswar' is a smokeless tobacco product (STP) widely used in Pakistan. It has been correlated with oral and oesophageal cancer in recent clinical studies. The toxic effects associated with STPs have been associated with trace level contaminants present in these products. The toxin levels of Pakistani naswar are reported for the first time in this study. METHODS: A total of 30 Pakistani brands of naswar were tested for a variety of toxic constituents and carcinogens such as cadmium, arsenic, lead and other carcinogenic metals, nitrite and nitrate, and nicotine and pH. RESULTS: The average values of all the toxins studied were well above their allowable limits, making the product a health risk for consumers. Calculated lifetime cancer risk from cadmium and lead was 1 lac (100,000) to 10 lac (1,000,000) times higher than the minimum 10E-4 (0.00001) to 10E-6 (0.000001), which is the 'target range' for potentially hazardous substances, according to the US Environmental Protection Agency. Similarly, the level of arsenic was in the range of 0.15 to 14.04 MUg/g, the average being 1.25 MUg/g. The estimated average bioavailable concentration of arsenic is 0.125-0.25 MUg/g, which is higher than the allowable standard of 0.01 MUg/g. Similarly, the average minimum daily intake of chromium and nickel was 126.97 MUg and 122.01 MUg, as compared to allowable 30-35 MUg and 35 MUg, respectively; a 4-5 times higher exposure. However, beryllium was not detected in any of the brands studied. The pH was highly basic, averaging 8.56, which favours the formation of tobacco specific amines thus making the product potentially toxic. This study validates clinical studies correlating incidence of cancer with naswar use in Pakistan. CONCLUSIONS: This study shows that the production, packaging, sale and consumption of naswar should be regulated so as to protect the public from the health hazards associated with its consumption. PMID- 21642446 TI - Optimising surgical training: use of feedback to reduce errors during a simulated surgical procedure. AB - OBJECTIVE: To assess the effect of proximate or immediate feedback during an intensive training session. The authors hypothesised that provision of feedback during a training session would improve performance and learning curves. METHODS: Twenty-eight trainee surgeons participated in the study between September and December 2008. They were consecutively assigned to group 1 (n=16, no feedback) or group 2 (n=12, feedback) All the participants performed five hand-assisted laparoscopic colectomy procedures on the ProMIS surgical simulator. Efficiency of instrument use (instrument path length and smoothness) and predefined intraoperative error scores were assessed. Facilitators assisted their performance and answered questions when asked. Group 1 participants were given no extra assistance, but group 2 participants received standardised feedback and the chance to review errors after every procedure. Data were analysed using SPSS V.15. Mann-Whitney U tests were used to compare mean performance results, and analysis of variance was used to calculate within-subject improvement. RESULTS: Group 1 achieved better results for instrument path length (23 874 mm vs 39 086 mm, p=0.001) and instrument smoothness (2015 vs 2567, p=0.045) However, group 2 (feedback) performed significantly better with regard to error scores (14 vs 4.42, p=0.000). In addition, they demonstrated a smoother learning curve. Inter rater reliability for the error scores was 0.97. CONCLUSION: The provision of standardised proximate feedback was associated with significantly fewer errors and an improved learning curve. Reducing errors in the skills lab environment should lead to safer clinical performance. This may help to make training more efficient and improve patient safety. PMID- 21642447 TI - HIV-1 Nef impairs multiple T-cell functions in antigen-specific immune response in mice. AB - The viral protein Nef is a key element for the progression of HIV disease. Previous in vitro studies suggested that Nef expression in T-cell lines enhanced TCR signaling pathways upon stimulation with TCR cross-linking, leading to the proposal that Nef lowers the threshold of T-cell activation, thus increasing susceptibility to viral replication in immune response. Likewise, the in vivo effects of Nef transgenic mouse models supported T-cell hyperresponse by Nef. However, the interpretation is complicated by Nef expression early in the development of T cells in these animal models. Here, we analyzed the consequence of Nef expression in ovalbumin-specific/CD4(+) peripheral T cells by using a novel mouse model and demonstrate that Nef inhibits antigen-specific T-cell proliferation and multiple functions required for immune response in vivo, which includes T-cell helper activity for the primary and memory B-cell response. However, Nef does not completely abrogate T-cell activity, as defined by low levels of cytokine production, which may afford the virus a replicative advantage. These results support a model, in which Nef expression does not cause T-cell hyperresponse in immune reaction, but instead reduces the T-cell activity, that may contribute to a low level of virus spread without viral cytopathic effects. PMID- 21642448 TI - Increased folding and channel activity of a rare cystic fibrosis mutant with CFTR modulators. AB - Cystic fibrosis (CF) is a lethal recessive genetic disease caused by mutations in the CFTR gene. The gene product is a PKA-regulated anion channel that is important for fluid and electrolyte transport in the epithelia of lung, gut, and ducts of the pancreas and sweat glands. The most common CFTR mutation, DeltaF508, causes a severe, but correctable, folding defect and gating abnormality, resulting in negligible CFTR function and disease. There are also a large number of rare CF-related mutations where disease is caused by CFTR misfolding. Yet the extent to which defective biogenesis of these CFTR mutants can be corrected is not clear. CFTRV232D is one such mutant that exhibits defective folding and trafficking. CFTRDeltaF508 misfolding is difficult to correct, but defective biogenesis of CFTRV232D is corrected to near wild-type levels by small-molecule folding correctors in development as CF therapeutics. To determine if CFTRV232D protein is competent as a Cl(-) channel, we utilized single-channel recordings from transfected human embryonic kidney (HEK-293) cells. After PKA stimulation, CFTRV232D channels were detected in patches with a unitary Cl(-) conductance indistinguishable from that of CFTR. Yet the frequency of detecting CFTRV232D channels was reduced to ~20% of patches compared with 60% for CFTR. The folding corrector Corr-4a increased the CFTRV232D channel detection rate and activity to levels similar to CFTR. CFTRV232D-corrected channels were inhibited with CFTR(inh 172) and stimulated fourfold by the CFTR channel potentiator VRT-532. These data suggest that CF patients with rare mutations that cause CFTR misfolding, such as CFTRV232D, may benefit from treatment with folding correctors and channel potentiators in development to restore CFTRDeltaF508 function. PMID- 21642449 TI - Mechanical stimuli and IL-13 interact at integrin adhesion complexes to regulate expression of smooth muscle myosin heavy chain in airway smooth muscle tissue. AB - Airway smooth muscle phenotype may be modulated in response to external stimuli under physiological and pathophysiological conditions. The effect of mechanical forces on airway smooth muscle phenotype were evaluated in vitro by suspending weights of 0.5 or 1 g from the ends of canine tracheal smooth muscle tissues, incubating the weighted tissues for 6 h, and then measuring the expression of the phenotypic marker protein, smooth muscle myosin heavy chain (SmMHC). Incubation of the tissues at a high load significantly increased expression of SmMHC compared with incubation at low load. Incubation of the tissues at a high load also decreased activation of PKB/Akt, as indicated by its phosphorylation at Ser 473. Inhibition of Akt or phosphatidylinositol-3,4,5 triphosphate-kinase increased SmMHC expression in tissues at low load but did not affect SmMHC expression at high load. IL-13 induced a significant increase in Akt activation and suppressed the expression of SmMHC protein at both low and high loads. The role of integrin signaling in mechanotransduction was evaluated by expressing a PINCH (LIM1-2) fragment in the muscle tissues that prevents the membrane localization of the integrin-binding IPP complex (ILK/PINCH/alpha-parvin), and also by expressing an inactive integrin-linked kinase mutant (ILK S343A) that inhibits endogenous ILK activity. Both mutants inhibited Akt activation and increased expression of SmMHC protein at low load but had no effect at high load. These results suggest that mechanical stress and IL-13 both act through an integrin-mediated signaling pathway to oppositely regulate the expression of phenotypic marker proteins in intact airway smooth muscle tissues. The stimulatory effects of mechanical stress on contractile protein expression oppose the suppression of contractile protein expression mediated by IL-13; thus the imposition of mechanical strain may inhibit changes in airway smooth muscle phenotype induced by inflammatory mediators. PMID- 21642450 TI - Genome sequence of the repetitive-sequence-rich Mycoplasma fermentans strain M64. AB - Mycoplasma fermentans is a microorganism commonly found in the genitourinary and respiratory tracts of healthy individuals and AIDS patients. The complete genome of the repetitive-sequence-rich M. fermentans strain M64 is reported here. Comparative genomics analysis revealed dramatic differences in genome size between this strain and the recently completely sequenced JER strain. PMID- 21642451 TI - The Campylobacter jejuni transcriptional regulator Cj1556 plays a role in the oxidative and aerobic stress response and is important for bacterial survival in vivo. AB - Campylobacter jejuni is the leading bacterial cause of human gastroenteritis worldwide. Despite stringent microaerobic growth requirements, C. jejuni is ubiquitous in the aerobic environment and so must possess regulatory systems to sense and adapt to external stimuli, such as oxidative and aerobic (O(2)) stress. Reannotation of the C. jejuni NCTC11168 genome sequence identified Cj1556 (originally annotated as a hypothetical protein) as a MarR family transcriptional regulator, and further analysis indicated a potential role in regulating the oxidative stress response. A C. jejuni 11168H Cj1556 mutant exhibited increased sensitivity to oxidative and aerobic stress, decreased ability for intracellular survival in Caco-2 human intestinal epithelial cells and J774A.1 mouse macrophages, and a reduction in virulence in the Galleria mellonella infection model. Microarray analysis of gene expression changes in the Cj1556 mutant indicated negative autoregulation of Cj1556 expression and downregulation of genes associated with oxidative and aerobic stress responses, such as katA, perR, and hspR. Electrophoretic mobility shift assays confirmed the binding of recombinant Cj1556 to the promoter region upstream of the Cj1556 gene. cprS, which encodes a sensor kinase involved in regulation of biofilm formation, was also upregulated in the Cj1556 mutant, and subsequent studies showed that the mutant had a reduced ability to form biofilms. This study identified a novel C. jejuni transcriptional regulator, Cj1556, that is involved in oxidative and aerobic stress responses and is important for the survival of C. jejuni in the natural environment and in vivo. PMID- 21642452 TI - Genome sequence of the mercury-methylating and pleomorphic Desulfovibrio africanus Strain Walvis Bay. AB - Desulfovibrio africanus strain Walvis Bay is an anaerobic sulfate-reducing bacterium capable of producing methylmercury (MeHg), a potent human neurotoxin. The mechanism of methylation by this and other organisms is unknown. We present the 4.2-Mb genome sequence to provide further insight into microbial mercury methylation and sulfate-reducing bacteria. PMID- 21642453 TI - Identification of an anchor residue for CheA-CheY interactions in the chemotaxis system of Escherichia coli. AB - Transfer of a phosphoryl group from autophosphorylated CheA (P-CheA) to CheY is an important step in the bacterial chemotaxis signal transduction pathway. This reaction involves CheY (i) binding to the P2 domain of P-CheA and then (ii) acquiring the phosphoryl group from the P1 domain. Crystal structures indicated numerous side chain interactions at the CheY-P2 binding interface. To investigate the individual contributions of the P2 side chains involved in these contacts, we analyzed the effects of eight alanine substitution mutations on CheA-CheY binding interactions. An F214A substitution in P2 caused ~1,000-fold reduction in CheA CheY binding affinity, while Ala substitutions at other P2 positions had small effects (E171A, E178A, and I216A) or no detectable effects (H181A, D202A, D207A, and C213A) on binding affinity. These results are discussed in relation to previous in silico predictions of hot-spot and anchor positions at the CheA-CheY interface. We also investigated the consequences of these mutations for chemotaxis signal transduction in living cells. CheA(F214A) was defective in mediating localization of CheY-YFP to the large clusters of signaling proteins that form at the poles of Escherichia coli cells, while the other CheA variants did not differ from wild-type (wt) CheA (CheA(wt)) in this regard. In our set of mutants, only CheA(F214A) exhibited a markedly diminished ability to support chemotaxis in motility agar assays. Surprisingly, however, in FRET assays that monitored receptor-regulated production of phospho-CheY, CheA(F214A) (and each of the other Ala substitution mutants) performed just as well as CheA(wt). Overall, our findings indicate that F214 serves as an anchor residue at the CheA-CheY interface and makes an important contribution to the binding energy in vitro and in vivo; however, loss of this contribution does not have a large negative effect on the overall ability of the signaling pathway to modulate P-CheY levels in response to chemoattractants. PMID- 21642454 TI - Regulation of flagellum number by FliA and FlgM and role in biofilm formation by Rhodobacter sphaeroides. AB - The FlgM secretion checkpoint plays a crucial role in coordinating bacterial flagellar assembly. Here we identify a new role for FlgM and FliA as part of a complex regulatory network which controls flagellum number and is essential for efficient swimming and biofilm formation in the monotrichous bacterium Rhodobacter sphaeroides. PMID- 21642455 TI - Structure-guided investigation of lipopolysaccharide O-antigen chain length regulators reveals regions critical for modal length control. AB - The O-antigen component of the lipopolysaccharide (LPS) represents a population of polysaccharide molecules with nonrandom (modal) chain length distribution. The number of the repeat O units in each individual O-antigen polymer depends on the Wzz chain length regulator, an inner membrane protein belonging to the polysaccharide copolymerase (PCP) family. Different Wzz proteins confer vastly different ranges of modal lengths (4 to >100 repeat units), despite having remarkably conserved structural folds. The molecular mechanism responsible for the selective preference for a certain number of O units is unknown. Guided by the three-dimensional structures of PCPs, we constructed a panel of chimeric molecules containing parts of two closely related Wzz proteins from Salmonella enterica and Shigella flexneri which confer different O-antigen chain length distributions. Analysis of the O-antigen length distribution imparted by each chimera revealed the region spanning amino acids 67 to 95 (region 67 to 95), region 200 to 255, and region 269 to 274 as primarily affecting the length distribution. We also showed that there is no synergy between these regions. In particular, region 269 to 274 also influenced chain length distribution mediated by two distantly related PCPs, WzzB and FepE. Furthermore, from the 3 regions uncovered in this study, region 269 to 274 appeared to be critical for the stability of the oligomeric form of Wzz, as determined by cross-linking experiments. Together, our data suggest that chain length determination depends on regions that likely contribute to stabilize a supramolecular complex. PMID- 21642456 TI - Residues in a conserved alpha-helical segment are required for cleavage but not secretion of an Escherichia coli serine protease autotransporter passenger domain. AB - Autotransporters are a superfamily of virulence factors produced by Gram-negative bacteria that are comprised of an N-terminal extracellular domain (passenger domain) and a C-terminal beta barrel domain (beta domain) that resides in the outer membrane (OM). The beta domain promotes the translocation of the passenger domain across the OM by an unknown mechanism. Available evidence indicates that an alpha-helical segment that spans the passenger domain-beta domain junction is embedded inside the beta domain at an early stage of assembly. Following its secretion, the passenger domain of the serine protease autotransporters of the Enterobacteriaceae (SPATEs) and the pertactin family of Bordetella pertussis autotransporters is released from the beta domain through an intrabarrel autoproteolytic cleavage of the alpha-helical segment. Although the mutation of conserved residues that surround the cleavage site has been reported to impair both the translocation and cleavage of the passenger domain of a SPATE called Tsh, we show here that the mutation of the same residues in another SPATE (EspP) affects only passenger domain cleavage. Our results strongly suggest that the conserved residues are required to position the alpha-helical segment for the cleavage reaction and are not required to promote passenger domain secretion. PMID- 21642457 TI - Physiological roles of the cyAbrB transcriptional regulator pair Sll0822 and Sll0359 in Synechocystis sp. strain PCC 6803. AB - All known cyanobacterial genomes possess multiple copies of genes encoding AbrB like transcriptional regulators, known as cyAbrBs, which are distinct from those conserved among other bacterial species. In this study, we addressed the physiological roles of Sll0822 and Sll0359, the two cyAbrBs in Synechocystis sp. strain PCC 6803, under nonstress conditions (20 MUmol of photons m-2 s-1 in ambient CO2). When the sll0822 gene was disrupted, the expression levels of nitrogen-related genes such as urtA, amt1, and glnB significantly decreased compared with those in the wild-type cells. Possibly due to the increase of the cellular carbon/nitrogen ratio in the sll0822-disrupted cells, a decrease in pigment contents, downregulation of carbon-uptake related genes, and aberrant accumulation of glycogen took place. Moreover, the mutant exhibited the decrease in the expression level of cytokinesis-related genes such as ftsZ and ftsQ, resulting in the defect in cell division and significant increase in cell size. The pleiotrophic phenotype of the mutant was efficiently suppressed by the introduction of Sll0822 and also partially suppressed by the introduction of Sll0359. When His-tagged cyAbrBs were purified from overexpression strains, Sll0359 and Sll0822 were copurified with each other. The cyAbrBs in Synechocystis sp. strain PCC 6803 seem to interact with each other and regulate carbon and nitrogen metabolism as well as the cell division process under nonstress conditions. PMID- 21642458 TI - Wall teichoic Acid-dependent adsorption of staphylococcal siphovirus and myovirus. AB - The molecular interactions between staphylococcal phages and host cell surfaces are poorly understood. Employing Staphylococcus aureus teichoic acid mutants, we demonstrate that wall teichoic acid (WTA), but not lipoteichoic acid, serves as a receptor for staphylococcal siphovirus and myovirus, while only the siphovirus requires glycosylated WTA. PMID- 21642459 TI - Characterization of the self-cleaving effector protein NopE1 of Bradyrhizobium japonicum. AB - NopE1 is a type III-secreted protein of the symbiont Bradyrhizobium japonicum which is expressed in nodules. In vitro it exhibits self-cleavage in a duplicated domain of unknown function (DUF1521) but only in the presence of calcium. Here we show that either domain is self-sufficient for cleavage. An exchange of the aspartic acid residue at the cleavage site with asparagine prevented cleavage; however, cleavage was still observed with glutamic acid at the same position, indicating that a negative charge at the cleavage site is sufficient. Close to each cleavage site, an EF-hand-like motif is present. A replacement of one of the conserved aspartic acid residues with alanine prevented cleavage at the neighboring site. Except for EDTA, none of several protease inhibitors blocked cleavage, suggesting that a known protease-like mechanism is not involved in the reaction. In line with this, the reaction takes place within a broad pH and temperature range. Interestingly, magnesium, manganese, and several other divalent cations did not induce cleavage, indicating a highly specific calcium binding site. Based on results obtained by blue-native gel electrophoresis, it is likely that the uncleaved protein forms a dimer and that the fragments of the cleaved protein oligomerize. A database search reveals that the DUF1521 domain is present in proteins encoded by Burkholderia phytofirmans PsNJ (a plant growth promoting betaproteobacterium) and Vibrio coralliilyticus ATCC BAA450 (a pathogenic gammaproteobacterium). Obviously, this domain is more widespread in proteobacteria, and it might contribute to the interaction with hosts. PMID- 21642460 TI - Multiplicity of 3-Ketosteroid-9alpha-Hydroxylase enzymes in Rhodococcus rhodochrous DSM43269 for specific degradation of different classes of steroids. AB - The well-known large catabolic potential of rhodococci is greatly facilitated by an impressive gene multiplicity. This study reports on the multiplicity of kshA, encoding the oxygenase component of 3-ketosteroid 9alpha-hydroxylase, a key enzyme in steroid catabolism. Five kshA homologues (kshA1 to kshA5) were previously identified in Rhodococcus rhodochrous DSM43269. These KshA(DSM43269) homologues are distributed over several phylogenetic groups. The involvement of these KshA homologues in the catabolism of different classes of steroids, i.e., sterols, pregnanes, androstenes, and bile acids, was investigated. Enzyme activity assays showed that all KSH enzymes with KshA(DSM43269) homologues are C 9 alpha-hydroxylases acting on a wide range of 3-ketosteroids, but not on 3 hydroxysteroids. KshA5 appeared to be the most versatile enzyme, with the broadest substrate range but without a clear substrate preference. In contrast, KshA1 was found to be dedicated to cholic acid catabolism. Transcriptional analysis and functional complementation studies revealed that kshA5 supported growth on any of the different classes of steroids tested, consistent with its broad expression induction pattern. The presence of multiple kshA genes in the R. rhodochrous DSM43269 genome, each displaying unique steroid induction patterns and substrate ranges, appears to facilitate a dynamic and fine-tuned steroid catabolism, with C-9 alpha-hydroxylation occurring at different levels during microbial steroid degradation. PMID- 21642461 TI - Structure of the flagellar motor protein complex PomAB: implications for the torque-generating conformation. AB - The bacterial flagellar motor is driven by an ion flux through a channel called MotAB in Escherichia coli or Salmonella and PomAB in Vibrio alginolyticus. PomAB is composed of two transmembrane (TM) components, PomA and PomB, and converts a sodium ion flux to rotation of the flagellum. Its homolog, MotAB, utilizes protons instead of sodium ions. PomB/MotB has a peptidoglycan (PG)-binding motif in the periplasmic domain, allowing it to function as the stator by being anchored to the PG layer. To generate torque, PomAB/MotAB is thought to undergo a conformational change triggered by the ion flux and to interact directly with FliG, a component of the rotor. Here, we present the first three-dimensional structure of this torque-generating stator unit analyzed by electron microscopy. The structure of PomAB revealed two arm domains, which contain the PG-binding site, connected to a large base made of the TM and cytoplasmic domains. The arms lean downward to the membrane surface, likely representing a "plugged" conformation, which would prevent ions leaking through the channel. We propose a model for how PomAB units are placed around the flagellar basal body to function as torque generators. PMID- 21642462 TI - Helicobacter pylori possesses four coiled-coil-rich proteins that form extended filamentous structures and control cell shape and motility. AB - We identified two additional genes of Helicobacter pylori encoding Ccrp proteins. All four Ccrps have different multimerization and filamentation properties and different types of smallest subunits and do not copurify, suggesting a system of individual Ccrp filaments. Despite the presence of morphologically unaltered flagella, all ccrp mutants displayed significantly reduced motility. PMID- 21642463 TI - Novel insights into the regulation of LexA in the cyanobacterium Synechocystis sp. Strain PCC 6803. AB - The transcription factor LexA in the cyanobacterium Synechocystis sp. strain PCC 6803 has been shown to regulate genes that are not directly involved in DNA repair but instead in several different metabolic pathways. However, the signal transduction pathways remain largely uncharacterized. The present work gives novel insights into the regulation of LexA in this unicellular cyanobacterium. A combination of Northern and Western blotting, using specific antibodies against the cyanobacterial LexA, was employed to show that this transcription regulator is under posttranscriptional control, in addition to the classical and already described transcriptional regulation. Moreover, detailed two-dimensional (2D) electrophoresis analyses of the protein revealed that LexA undergoes posttranslational modifications. Finally, a fully segregated LexA::GFP (green fluorescent protein) fusion-modified strain was produced to image LexA's spatial distribution in live cells. The fusion protein retains DNA binding capabilities, and the GFP fluorescence indicates that LexA is localized in the innermost region of the cytoplasm, decorating the DNA in an evenly distributed pattern. The implications of these findings for the overall role of LexA in Synechocystis sp. strain PCC 6803 are further discussed. PMID- 21642464 TI - Recognition of DNA by the helix-turn-helix global regulatory protein Lrp is modulated by the amino terminus. AB - The AsnC/Lrp family of regulatory proteins links bacterial and archaeal transcription patterns to metabolism. In Escherichia coli, Lrp regulates approximately 400 genes, over 200 of them directly. In earlier studies, lrp genes from Vibrio cholerae, Proteus mirabilis, and E. coli were introduced into the same E. coli background and yielded overlapping but significantly different regulons. These differences were seen despite amino acid sequence identities of 92% (Vibrio) and 98% (Proteus) to E. coli Lrp, including complete conservation of the helix-turn-helix motifs. The N-terminal region contains many of the sequence differences among these Lrp orthologs, which led us to investigate its role in Lrp function. Through the generation of hybrid proteins, we found that the N terminal diversity is responsible for some of the differences between orthologs in terms of DNA binding (as revealed by mobility shift assays) and multimerization (as revealed by gel filtration, dynamic light scattering, and analytical ultracentrifugation). These observations indicate that the N-terminal tail plays a significant role in modulating Lrp function, similar to what is seen for a number of other regulatory proteins. PMID- 21642465 TI - Acinetobacter baumannii RecA protein in repair of DNA damage, antimicrobial resistance, general stress response, and virulence. AB - RecA is the major enzyme involved in homologous recombination and plays a central role in SOS mutagenesis. In Acinetobacter spp., including Acinetobacter baumannii , a multidrug-resistant bacterium responsible for nosocomial infections worldwide, DNA repair responses differ in many ways from those of other bacterial species. In this work, the function of A. baumannii RecA was examined by constructing a recA mutant. Alteration of this single gene had a pleiotropic effect, showing the involvement of RecA in DNA damage repair and consequently in cellular protection against stresses induced by DNA damaging agents, several classes of antibiotics, and oxidative agents. In addition, the absence of RecA decreased survival in response to both heat shock and desiccation. Virulence assays in vitro (with macrophages) and in vivo (using a mouse model) similarly implicated RecA in the pathogenicity of A. baumannii . Thus, the data strongly suggest a protective role for RecA in the bacterium and indicate that inactivation of the protein can contribute to a combined therapeutic approach to controlling A. baumannii infections. PMID- 21642466 TI - Identification and characterization of a novel serine protease, VvpS, that contains two functional domains and is essential for autolysis of Vibrio vulnificus. AB - Little is known about the molecular mechanism for autolysis of Gram-negative bacteria. In the present study, we identified the vvpS gene encoding a serine protease, VvpS, from Vibrio vulnificus, a Gram-negative food-borne pathogen. The amino acid sequence predicted that VvpS consists of two functional domains, an N terminal protease catalytic domain (PCD) and a C-terminal carbohydrate binding domain (CBD). A null mutation of vvpS significantly enhanced viability during stationary phase, as measured by enumerating CFU and differentially staining viable cells. The vvpS mutant reduced the release of cytoplasmic beta galactosidase and high-molecular-weight extracellular chromosomal DNA into the culture supernatants, indicating that VvpS contributes to the autolysis of V. vulnificus during stationary phase. VvpS is secreted via a type II secretion system (T2SS), and it exerts its effects on autolysis through intracellular accumulation during stationary phase. Consistent with this, a disruption of the T2SS accelerated intracellular accumulation of VvpS and thereby the autolysis of V. vulnificus. VvpS also showed peptidoglycan-hydrolyzing activity, indicating that the autolysis of V. vulnificus is attributed to the self-digestion of the cell wall by VvpS. The functions of the VvpS domains were assessed by C-terminal deletion analysis and demonstrated that the PCD indeed possesses a proteolytic activity and that the CBD is required for hydrolyzing peptidoglycan effectively. Finally, the vvpS mutant exhibited reduced virulence in the infection of mice. In conclusion, VvpS is a serine protease with a modular structure and plays an essential role in the autolysis and pathogenesis of V. vulnificus. PMID- 21642467 TI - Identification of amino acid residues critical for catalysis of Holliday junction resolution by Mycoplasma genitalium RecU. AB - The RecU protein from Mycoplasma genitalium, RecU(Mge), is a 19.4-kDa Holliday junction (HJ) resolvase that binds in a nonspecific fashion to HJ substrates and, in the presence of Mn(2+), cleaves these substrates at a specific sequence (5' G/TC?C/TTA/GG-3'). To identify amino acid residues that are crucial for HJ binding and/or cleavage, we generated a series of 16 deletion mutants (9 N- and 7 C-terminal deletion mutants) and 31 point mutants of RecU(Mge). The point mutations were introduced at amino acid positions that are highly conserved among bacterial RecU-like sequences. All mutants were purified and tested for the ability to bind to, and cleave, HJ substrates. We found the five N-terminal and three C-terminal amino acid residues of RecU(Mge) to be dispensable for its catalytic activities. Among the 31 point mutants, 7 mutants were found to be inactive in both HJ binding and cleavage. Interestingly, in 12 other mutants, these two activities were uncoupled; while these proteins displayed HJ-binding characteristics similar to those of wild-type RecU(Mge), they were unable to cleave HJ substrates. Thus, 12 amino acid residues were identified (E11, K31, D57, Y58, Y66, D68, E70, K72, T74, K76, Q88, and L92) that may play either a direct or indirect role in the catalysis of HJ resolution. PMID- 21642468 TI - Complete genome sequence of the anaerobic, halophilic alkalithermophile Natranaerobius thermophilus JW/NM-WN-LF. AB - The genome of the anaerobic halophilic alkalithermophile Natranaerobius thermophilus consists of one 3,165,557-bp chromosome and two plasmids (17,207 bp and 8,689 bp). The present study is the first to report the completely sequenced genome of an anaerobic polyextremophile and genes associated with roles in regulation of intracellular osmotic pressure, pH homeostasis, and growth at elevated temperatures. PMID- 21642469 TI - Malnutrition causing neonatal dyslipidemia. AB - Lipid abnormalities in children have become more common in recent decades. This trend is related to the increase in overweight and obesity. The 2002 National Health and Nutrition Examination Survey reported that the percentage of risk for overweight and overweight in children aged > 6 years is 31%, higher than the previous surveys. Serum lipids tend to increase quickly up to 6 months of age and reach values very close to adult values by age 2. As suggested by the American Heart Association, serum lipid values for children and adolescents (2-19 years old) are considered abnormal when total cholesterol is >200 mg/dL, high-density lipoprotein is <35 mg/dL, low-density lipoprotein is >130 mg/dL, and triglycerides are >150 mg/dL. Dyslipidemia can be found in patients with malnutrition, a severe condition that needs prompt nutrition intervention. This report describes a case of malnutrition causing severe dyslipidemia in a newborn. Primary dyslipidemia was excluded by the presence of primary malnutrition, normal response to a postheparin lipoprotein lipase activity test, a favorable clinical course after nutrition intervention, and relatives' blood lipid levels close to normal that did not indicate familial dyslipidemia. The child was fed fat-free milk formula supplemented with medium-chain triglycerides and had adequate weight gain with a decrease in blood lipids. Subsequently the formula was changed to regular milk-based formula, and the child maintained adequate growth rate. Although blood lipids never returned to normal values for age and sex, they were lower than before treatment. PMID- 21642470 TI - Lack of significant effect of bilastine administered at therapeutic and supratherapeutic doses and concomitantly with ketoconazole on ventricular repolarization: results of a thorough QT study (TQTS) with QT-concentration analysis. AB - The effect of bilastine on cardiac repolarization was studied in 30 healthy participants during a multiple-dose, triple-dummy, crossover, thorough QT study that included 5 arms: placebo, active control (400 mg moxifloxacin), bilastine at therapeutic and supratherapeutic doses (20 mg and 100 mg once daily, respectively), and bilastine 20 mg administered with ketoconazole 400 mg. Time matched, triplicate electrocardiograms (ECGs) were recorded with 13 time points extracted predose and 16 extracted over 72 hours post day 4 dosing. Four QT/RR corrections were implemented: QTcB; QTcF; a linear individual correction (QTcNi), the primary correction; and a nonlinear one (QTcNnl). Moxifloxacin was associated with a significant increase in QTcNi at all time points between 1 and 12 hours, inclusively. Bilastine administration at 20 mg and 100 mg had no clinically significant impact on QTc (maximum increase in QTcNi, 5.02 ms; upper confidence limit [UCL] of the 1-sided, 95% confidence interval, 7.87 ms). Concomitant administration of ketoconazole and bilastine 20 mg induced a clinically relevant increase in QTc (maximum increase in QTcNi, 9.3 ms; UCL, 12.16 ms). This result was most likely related to the cardiac effect of ketoconazole because for all time points, bilastine plasma concentrations were lower than those observed following the supratherapeutic dose. PMID- 21642471 TI - Human polymorphisms in the glutathione transferase zeta 1/maleylacetoacetate isomerase gene influence the toxicokinetics of dichloroacetate. AB - Dichloroacetate (DCA), a chemical relevant to environmental science and allopathic medicine, is dehalogenated by the bifunctional enzyme glutathione transferase zeta (GSTz1)/maleylacetoacetate isomerase (MAAI), the penultimate enzyme in the phenylalanine/tyrosine catabolic pathway. The authors postulated that polymorphisms in GSTz1/MAAI modify the toxicokinetics of DCA. GSTz1/MAAI haplotype significantly affected the kinetics and biotransformation of 1,2-13C DCA when it was administered at either environmentally (ug/kg/d) or clinically (mg/kg/d) relevant doses. GSTz1/MAAI haplotype also influenced the urinary accumulation of potentially toxic tyrosine metabolites. Atomic modeling revealed that GSTz1/MAAI variants associated with the slowest rates of DCA metabolism induced structural changes in the enzyme homodimer, predicting protein instability or abnormal protein-protein interactions. Knowledge of the GSTz1/MAAI haplotype can be used prospectively to identify individuals at potential risk of DCA's adverse side effects from environmental or clinical exposure or who may exhibit aberrant amino acid metabolism in response to dietary protein. PMID- 21642472 TI - Role of von Hippel-Lindau protein in fibroblast proliferation and fibrosis. AB - Idiopathic pulmonary fibrosis (IPF) is characterized by exaggerated fibroblast proliferation and accumulation of collagens and fibronectin. The extracellular fibronectin and collagen network is regulated by von Hippel-Lindau protein (pVHL). However, it is unknown whether pVHL contributes to pulmonary fibrosis. We found that lungs from patients with IPF expressed increased levels of pVHL in fibroblastic foci. Bleomycin treatment also induced pVHL in lung fibroblasts, but not in alveolar type II cells. Overexpression of pVHL increased lung fibroblast proliferation, protein abundance of fibronectin and collagen, and extracellular fibronectin. In addition, overexpression of pVHL induced expression of the alpha5 integrin subunit. Overexpression of pVHL did not alter hypoxia-inducible factor luciferase reporter activity and mRNA expression of vascular endothelial growth factor. Fibroblasts overexpressing pVHL were more sensitive to RGD peptide mediated reduction in proliferation. Activating alpha5 and beta1 integrin increased proliferation of fibroblasts overexpressing pVHL and those cells were more resistant to the inhibition of alpha5 integrin. Overexpression of pVHL also increased activation of focal adhesion kinase (FAK). Moreover, suppression of pVHL prevented TGF-beta1-induced proliferation of mouse embryonic fibroblasts. Taken together, our results indicate that elevated expression of pVHL results in the aberrant fibronectin expression, activation of integrin/FAK signaling, fibroblast proliferation, and fibrosis. PMID- 21642473 TI - Deletion of Zfp521 rescues the growth plate phenotype in a mouse model of Jansen metaphyseal chondrodysplasia. AB - Jansen metaphyseal chondrodysplasia (JMC) is caused by a constitutively activating mutation of the parathyroid hormone (PTH)/PTH-related protein (PTHrP) receptor (PTHR1) and is characterized by widening of the metaphyses, reduction of long bone length, and short stature. A transgenic mouse expressing this mutation under the collagen alpha1(II) promoter has been generated to investigate the mechanisms responsible for this chondrodysplasia. We recently identified zinc finger protein 521 (Zfp521) as a downstream target gene of PTHrP signaling. Interestingly, loss of Zfp521 from chondrocytes leads to reduced cell proliferation and increased differentiation in the growth plate. Thus, we hypothesized that specifically ablating Zfp521 from Jansen chondrocytes could sufficiently rescue the chondrodysplasia phenotype. Our results show that Zfp521 expression is up-regulated in Jansen mouse growth plate chondrocytes and that PTHR1 is required for Zfp521 expression. Its ablation from Jansen chondrocytes restored normal cell differentiation, thus initiating chondrocyte apoptosis at the chondro-osseous junction, leading to partial rescue of endochondral bone formation shown by proper bone length. This study provides the first genetic evidence that Zfp521 is required downstream of PTHR1 signaling to act on chondrocyte proliferation, differentiation, and cell death. PMID- 21642474 TI - Tyrosine phosphorylation of transcriptional coactivator WW-domain binding protein 2 regulates estrogen receptor alpha function in breast cancer via the Wnt pathway. AB - WW-binding protein 2 (WBP2) has been demonstrated in different studies to be a tyrosine kinase substrate, to activate estrogen receptor alpha (ERalpha)/progesterone receptor (PR) transcription, and to play a role in breast cancer. However, the role of WBP2 tyrosine phosphorylation in regulating ERalpha function and breast cancer biology is unknown. Here, we established WBP2 as a tyrosine phosphorylation target of estrogen signaling via EGFR crosstalk. Using dominant-negative, constitutively active mutants, RNAi, and pharmacological studies, we demonstrated that phosphorylation of WBP2 at Tyr192 and Tyr231 could be regulated by c-Src and c-Yes kinases. We further showed that abrogating WBP2 phosphorylation impaired >60% of ERalpha reporter activity, putatively by blocking nuclear entry of WBP2 and its interaction with ERalpha. Compared to vector control, overexpression of WBP2 and its phospho-mimic mutant in MCF7 cells resulted in larger tumors in mice, induced loss of cell-cell adhesion, and enhanced cell proliferation, anchorage-independent growth, migration, and invasion in both estrogen-dependent and -independent manners, events of which could be substantially abolished by overexpression of the phosphorylation defective mutant. Hormone independence of cells expressing WBP2 phospho-mimic mutant was associated with heightened ERalpha and Wnt reporter activities. Wnt/beta-catenin inhibitor FH535 blocked phospho-WBP2-mediated cancer cell growth more pronouncedly than tamoxifen and fulvestrant, in part by reducing the expression of ERalpha. Wnt pathway is likely to be a critical component in WBP2 mediated breast cancer biology. PMID- 21642475 TI - Police job strain during routine activities and a major event. AB - BACKGROUND: Special police forces are exposed to periods of intense work stress in ensuring public order. AIMS: To explore the relationship between the work context (routine work or special event) of special force policemen and psychological measures of job strain (demand-control) and effort-reward imbalance. METHODS: All policemen assigned to the G8 meeting in L'Aquila, Italy, in July 2009 were invited to complete a questionnaire while engaged in routine work in January 2009 (Time A) and in June 2009 (Time B), while preparing for the special event. RESULTS: Participation rate in the questionnaire study was 292/294 (99%) members of the special police force. Measures of job strain (-0.39, P < 0.001) and effort-reward imbalance (-0.37, P < 0.001) decreased significantly from Time A to Time B. On average, demand decreased from 14.2 +/- 1.9 to 12.6 +/- 2.7 (P < 0.001), control increased from 11.8 +/- 2.5 to 14.4 +/- 3.4 (P < 0.001) and social support increased from 17.8 +/- 2.9 to 19.0 +/- 3.1 (P < 0.001). At the same time, effort decreased from 17.4 +/- 3.2 to 11.8 +/- 3.8 (P < 0.001), reward grew from 37.6 +/- 5.5 to 45.5 +/- 7.4 (P < 0.001) and overcommitment dropped from 7.1 +/- 2.1 to 6.6 +/- 1.7 (P < 0.001). CONCLUSIONS: In special police forces, routine work may be significantly more stressful than a single critical event. PMID- 21642476 TI - Remission of a mother's depression is associated with her child's mental health. PMID- 21642477 TI - Stress and addiction. PMID- 21642478 TI - An amygdala structural abnormality common to two subtypes of conduct disorder: a neurodevelopmental conundrum. PMID- 21642479 TI - Cracking the code: dopamine signaling in cocaine dependence. PMID- 21642480 TI - Misdiagnosed postpartum psychosis revealing a late-onset urea cycle disorder. PMID- 21642481 TI - Studying the efficacy of psychodynamic psychotherapy. PMID- 21642482 TI - The short-term effect of the recent great earthquake on the general functioning status of psychiatric patients near Tokyo. PMID- 21642483 TI - Question regarding ziprasidone and QTc interval prolongation in the ZODIAC study. PMID- 21642484 TI - Asenapine for the treatment of stuttering: an analysis of three cases. PMID- 21642485 TI - Rhodococcus artemisiae sp. nov., an endophytic actinobacterium isolated from the pharmaceutical plant Artemisia annua L. AB - A Gram-positive, non-motile actinobacterium, designated YIM 65754T, was isolated from the stem of Artemisia annua L., collected from Yunnan province, south-west China. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain YIM 65754T comprised an evolutionary lineage within the genus Rhodococcus. The isolate clustered with Rhodococcus pyridinivorans PDB9T, Rhodococcus gordoniae W 4937T and Rhodococcus rhodochrous DSM 43241T, with which it shared 98.4, 97.9 and 97.8 % 16S rRNA gene sequence similarities, respectively. However, DNA-DNA relatedness demonstrated that strain YIM 65754T was distinct from its closest phylogenetic neighbours. The cell-wall peptidoglycan contained meso diaminopimelic acid, arabinose, galactose, mannose and glucose (cell-wall chemotype IV). The major menaquinone was MK-8(H2) and the predominant fatty acids were C16:0 (27.83 %), iso-C15:0 2-OH and/or C16:1omega7c (20.21 %) and 10-methyl C18:0 (17.50 %). The DNA G+C content was 66.2 mol%. On the basis of phenotypic, chemotaxonomic and phylogenetic evidence, the isolate represents a novel species of the genus Rhodococcus, for which the name Rhodococcus artemisiae sp. nov. is proposed; the type strain is YIM 65754T (=CCTCC AA 209042T=DSM 45380T). PMID- 21642486 TI - Planococcus plakortidis sp. nov., isolated from the marine sponge Plakortis simplex (Schulze). AB - A novel coccoid-shaped strain, AS/ASP6 (II)T, was isolated from a sample taken from Plakortis simplex (Schulze), a marine sponge, collected at a depth of 30 m from the Bay of Bengal. This strain was identified by using a polyphasic taxonomic approach. The results of 16S rRNA gene sequence analysis showed that strain AS/ASP6 (II)T should be assigned to the genus Planococcus. Chemotaxonomic data (A4alpha-type peptidoglycan; MK-6, MK-7 and MK-8 menaquinones; mainly branched cellular fatty acids; and phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol as cellular phospholipids) supported taxonomic placement in the genus Planococcus. Phylogenetic analysis of the 16S rRNA gene sequence showed that strain AS/ASP6 (II)T belonged to the genus Planococcus and was closely related to the type strains of Planococcus maritimus (99.1 %) followed by Planococcus rifietoensis (98.6 %), Planococcus maitriensis (98.5 %), Planococcus citreus (98.3 %), Planococcus salinarum (98.1 %), Planococcus columbae (97.9 %), Planococcus donghaensis (97.8 %) and Planococcus antarcticus (97.7 %); DNA-DNA hybridization values obtained were well below the threshold that is required for the proposal of a novel species. The G+C content of the genomic DNA was 51.0 mol%. The phenotypic and genotypic data showed that strain AS/ASP6 (II)T merits recognition as a representative of a novel species of the genus Planococcus, for which the name Planococcus plakortidis sp. nov. is proposed; the type strain is AS/ASP6 (II)T (=MTCC 8491T=DSM 23997T). PMID- 21642487 TI - Carboxydocella manganica sp. nov., a thermophilic, dissimilatory Mn(IV)- and Fe(III)-reducing bacterium from a Kamchatka hot spring. AB - A thermophilic, anaerobic, dissimilatory Mn(IV)- and Fe(III)-reducing bacterium (strain SLM 61T) was isolated from a terrestrial hot spring on the Kamchatka peninsula. The cells were straight rods, 0.5-0.6 um in diameter and 1.0-6.0 um long, and exhibited tumbling motility by means of peritrichous flagellation. The strain grew at 26-70 degrees C, with an optimum at 58-60 degrees C, and at pH 5.5-8.0, with an optimum at pH 6.5. Growth of SLM 61T was observed at 0-2.0 % (w/v) NaCl, with an optimum at 0.5 % (w/v). The generation time under optimal growth conditions was 40 min. Strain SLM 61T grew and reduced Mn(IV), Fe(III) or nitrate with a number of organic acids and complex proteinaceous compounds as electron donors. It was capable of chemolithoautotrophic growth using molecular hydrogen as an electron donor, Fe(III) but not Mn(IV) or nitrate as an electron acceptor and CO2 as a carbon source. It also was able to ferment pyruvate, yeast extract, glucose, fructose, sucrose and maltose. The G+C content of DNA of strain SLM 61T was 50.9 mol%. 16S rRNA gene sequence analysis revealed that the closest relative of the isolated organism was Carboxydocella thermautotrophica 41T (96.9 % similarity). On the basis of its physiological properties and phylogenetic analyses, the isolate is considered to represent a novel species, for which the name Carboxydocella manganica sp. nov. is proposed. The type strain is SLM 61T (=DSM 23132T=VKM B-2609T). C. manganica is the first described representative of the genus Carboxydocella that possesses the ability to reduce metals and does not utilize CO. PMID- 21642488 TI - Pseudomonas baetica sp. nov., a fish pathogen isolated from wedge sole, Dicologlossa cuneata (Moreau). AB - Five Gram-negative bacterial isolates, recovered from an outbreak that occurred in March 2006 in Huelva, Spain, affecting adult diseased cultured wedge sole [Dicologlossa cuneata (Moreau)], were characterized phenotypically and genotypically in order to clarify their taxonomic position. On the basis of 16S rRNA gene sequence analysis, the isolates were included in the genus Pseudomonas, within the Pseudomonas fluorescens-related species group, their closest relatives being the Pseudomonas jessenii and Pseudomonas koreensis subgroups. The highest sequence similarities were recorded with the type strains of Pseudomonas reinekei, P. moorei, P. umsongensis, P. jessenii and P. mohnii (99.4-99.3 % similarity). Sequence analysis of the housekeeping genes gyrB and rpoD clearly differentiated the isolates from currently described Pseudomonas species, the highest sequence similarities recorded to type strains being below 95 % for both genes. Phylogenetic analysis using concatenated sequences of the three genes showed Pseudomonas moraviensis DSM 16007T and P. koreensis DSM 16610T as the closest reference strains. DNA-DNA hybridization assays with related strains confirmed that these isolates belong to a novel species of the genus Pseudomonas, for which the name Pseudomonas baetica sp. nov. is proposed. The type strain is strain a390T (=CECT 7720T=LMG 25716T). The novel species could be easily distinguished from phylogenetically related species by several phenotypic characteristics, including gelatin hydrolysis, acid production from glucose and growth at 6 % NaCl. Virulence assays revealed that the novel species is pathogenic for wedge sole. PMID- 21642489 TI - Rhizobium skierniewicense sp. nov., isolated from tumours on chrysanthemum and cherry plum. AB - Three isolates of Gram-negative, rod-shaped, non-spore-forming bacteria were recovered from galls on chrysanthemum (Chrysanthemum L.; Ch11T, Ch12) and cherry plum (Prunus cerasifera var. divaricata; AL9.3). All three isolates were able to cause crown galls on various plant species. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the three isolates were probably identical (100% sequence similarity) and closely related to Rhizobium rubi (99.6 %), Rhizobium radiobacter (98.7 %) and Rhizobium larrymoorei (98.1 %). Similar analysis based on the housekeeping genes glnA, gyrB and rpoB also indicated that the novel isolates were identical and closely related to R. rubi. The major cellular fatty acids of strain Ch11T were C18:1omega7c (62.1 %), summed feature 2 (comprising C12:0 aldehyde, iso-C16:1 I and/or C14:0 3-OH; 10.8 %), summed feature 3 (comprising C16:1omega7c and/or iso-C15:0 2-OH; 7.7 %) and C10:0 3-OH (7.5 %). However, the DNA-DNA relatedness between Ch11T and R. rubi LMG 156T was only 48 % and, unlike phylogenetically related established Rhizobium species, the novel isolates were able to utilize beta-hydroxybutyric acid but not L-fucose. Based on the phylogenetic and phenotypic evidence, the isolates are considered to represent a single novel species of the genus Rhizobium, for which the name Rhizobium skierniewicense sp. nov. is proposed; the type strain is Ch11T (=LMG 26191T=CFBP 7420T). PMID- 21642490 TI - The influence of dietary lysine restriction during the finishing period on growth performance and carcass, meat, and fat characteristics of barrows and gilts intended for dry-cured ham production. AB - A total of 120 pigs [Duroc * (Landrace * Large White); initial average BW: 100.3 +/- 2.5 kg] were used to investigate the effects of sex (barrows and gilts) and dietary total Lys restriction (7.0, 6.5, and 6.0 g.kg(-1)) on growth performance and carcass, meat, and fat characteristics. Pigs were intended for high-quality dry-cured ham from Spain (called Teruel ham), and a minimum fat thickness at the gluteus medius muscle (GM) is required (16 mm) for carcasses to be acceptable. Animals were slaughtered when they reached 129.0 +/- 3.6 kg of BW. There were 6 treatments arranged factorially (2 sexes * 3 dietary Lys concentrations) and 4 replicates of 5 pigs per treatment. Barrows consumed more feed (P = 0.001) and tended to have less G:F (P = 0.06) than gilts. Carcasses from barrows were fatter (P = 0.001) and had heavier main trimmed lean cuts (P = 0.008) than gilts. A greater proportion of final acceptable carcasses for Teruel ham (P = 0.001) was observed in barrows than in gilts because of the greater percentage of carcasses that fulfill the minimum fat depth at GM required (P = 0.001). Meat from barrows had greater content of intramuscular fat (P = 0.02) than meat from gilts. Also, subcutaneous fat from barrows had less proportion of PUFA than fat from gilts (P = 0.02). A reduction in dietary Lys concentration decreased ADG (P = 0.004) and ADFI (P = 0.001) in pigs. In addition, backfat depth (P = 0.007) and fat at GM (P = 0.07) increased as dietary Lys decreased. The proportion of carcasses that fulfilled the minimum fat depth at GM required for Teruel ham increased as dietary Lys decreased in feed, but this effect was greater in gilts than in barrows (sex * Lys, P = 0.02). Meat and fat quality was not influenced by dietary treatment. We conclude that different feeding programs with different dietary Lys concentrations may be needed for barrows and gilts intended for production of dry cured hams where a minimum carcass fat depth is required. PMID- 21642491 TI - Cloning of comparative gene identification-58 gene in avian species and investigation of its developmental and nutritional regulation in chicken adipose tissue. AB - Adipose triglyceride lipase (ATGL) is the rate-limiting enzyme of lipolysis in chicken adipose tissue. Its regulation is not fully understood. Recent studies suggest ATGL may be regulated by physical protein-protein interactions. Comparative gene identification 58 (CGI-58) has been identified as an activator of ATGL in mice. The purpose of the current study was to clone and sequence the CGI-58 gene in avian species and to investigate its regulation during development, fasting, and refeeding. Here, we report the cloning and sequencing of the complete coding sequence of CGI-58 and the deduced AA sequences for the domestic chicken, turkey, and Coturnix quail. The CGI-58 protein is a 343-AA protein in the chicken and quail, and a 344-AA protein in the turkey. Sequence comparisons with the human and mouse show that the CGI-58 gene is highly conserved among avian and mammalian species, with complete identities at the predicted lipid-binding site. Cell fractionation of chicken fat cells and stromal vascular cells revealed that CGI-58 is expressed primarily in mature adipocytes (P < 0.01). When compared in multiple organs and tissues, avian CGI-58 is expressed predominantly in the adipose tissue (P < 0.001), similar to ATGL. To understand CGI-58 expression during adipose tissue development, its mRNA expression was measured along with ATGL and stearoyl CoA desaturase (SCD-1) mRNA, an adipogenic marker, in embryos and adults. Messenger RNA expression of CGI-58 increased (P < 0.05) immediately after hatching, concurrent with peak ATGL expression. It is interesting that CGI-58 remained somewhat increased at posthatch d 11 and 33 as SCD-1 mRNA expression increased (P < 0.05). To evaluate the response of CGI-58 to nutritional status, chickens and quail were fasted for 24 h and subsequently refed. After the fasting period, CGI-58 mRNA was induced (P < 0.05) for both chickens and quail and was returned to control levels upon refeeding. The ATGL mRNA responded similarly, increasing dramatically after fasting and quickly decreasing with refeeding. The direct relationship between CGI-58 and ATGL mRNA expression indicates a role for CGI-58 in activating ATGL mediated lipolysis in avian species. PMID- 21642492 TI - Effect of long-term corn by-product feeding on beef quality, strip loin fatty acid profiles, and shelf life. AB - The objective of this study was to evaluate the meat quality and shelf life of steaks from steers fed dried distillers grains with solubles (DDGS) or dried corn gluten feed (CGF) compared with soybean meal with corn (SBM) as a protein supplement from weaning to slaughter. Angus cross steers (n = 81; BW = 306 +/- 26.1 kg) were randomly assigned to pens (n = 9) and fed a stocker diet of corn silage (75% of DM) with DDGS, CGF, or SBM and ground ear corn. After 84 d of stockering, 12 steers (BW = 397 +/- 15.3 kg) were randomly selected from each treatment and finished using the same protein supplement at 25% of DM for 100 d. Carcass data were collected (24 h) and the longissimus lumborum was fabricated into steaks at 48 h postmortem. Steaks were assigned to proximate analysis, Warner-Bratzler shear force (7-, 14-, or 21-d aging), and retail display (1, 3, 6, or 9 d). Protein source did not affect carcass yield, quality, or longissimus lumborum composition (P > 0.05). After 7 d of aging, DDGS and CGF steaks were more tender (P < 0.01) than SBM, but were similar (P = 0.30) after 14 and 21 d of aging. Feeding corn by-products did not influence subjective overall color acceptance (P = 0.17) in this study, but acceptance declined over time (P < 0.01). Subjective redness was similar (P > 0.05) among diets except SBM steaks were more red (P < 0.01) than DDGS after 9 d. On d 3 and 6 of retail display, CGF steaks exhibited more discoloration (P < 0.04) than SBM or DDGS steaks. However, after 9 d DDGS steaks were more discolored (P < 0.01) than CGF or SBM. Objective L* was lighter for CGF (P < 0.04) over 9 d of display, and all treatments became darker (P < 0.01) as time increased. Redness (a*) declined (P < 0.01) over time with SBM steaks maintaining more color in the red spectrum than CGF and DDGS after 6 d of display. Protein source did not affect (P > 0.05) the rate of lipid oxidation. Total SFA concentrations were similar (P > 0.05) among treatments; however, total MUFA were less (P < 0.05) and total PUFA concentrations were greater (P < 0.05) in DDGS steaks compared with SBM or CGF steaks. These data show that DDGS or CGF can be fed as a protein supplement at 25% DM from weaning until slaughter while maintaining meat quality when compared with steers fed soybean meal as a protein supplement. PMID- 21642493 TI - Accuracy of genomic breeding values for residual feed intake in crossbred beef cattle. AB - The benefit of using genomic breeding values (GEBV) in predicting ADG, DMI, and residual feed intake for an admixed population was investigated. Phenotypic data consisting of individual daily feed intake measurements for 721 beef cattle steers tested over 5 yr was available for analysis. The animals used were an admixed population of spring-born steers, progeny of a cross between 3 sire breeds and a composite dam line. Training and validation data sets were defined by randomly splitting the data into training and testing data sets based on sire family so that there was no overlap of sires in the 2 sets. The random split was replicated to obtain 5 separate data sets. Two methods (BayesB and random regression BLUP) were used to estimate marker effects and to define marker panels and ultimately the GEBV. The accuracy of prediction (the correlation between the phenotypes and GEBV) was compared between SNP panels. Accuracy for all traits was low, ranging from 0.223 to 0.479 for marker panels with 200 SNP, and 0.114 to 0.246 for marker panels with 37,959 SNP, depending on the genomic selection method used. This was less than accuracies observed for polygenic EBV accuracies, which ranged from 0.504 to 0.602. The results obtained from this study demonstrate that the utility of genetic markers for genomic prediction of residual feed intake in beef cattle may be suboptimal. Differences in accuracy were observed between sire breeds when the random regression BLUP method was used, which may imply that the correlations obtained by this method were confounded by the ability of the selected SNP to trace breed differences. This may also suggest that prediction equations derived from such an admixed population may be useful only in populations of similar composition. Given the sample size used in this study, there is a need for increased feed intake testing if substantially greater accuracies are to be achieved. PMID- 21642494 TI - Associations of marker panel scores with feed intake and efficiency traits in beef cattle using preselected single nucleotide polymorphisms. AB - Because of the moderate heritability and the expense associated with collecting feed intake data, effective selection for residual feed intake would be enhanced if marker-assisted evaluation were used for accurate estimation of genetic merit. In this study, a suite of genetic markers predictive of residual feed intake, DMI, and ADG were preselected using single-marker regression analysis, and the top 100 SNP were analyzed further to provide prediction equations for the traits. The data used consisted of 728 spring-born beef steers, offspring of a cross between a composite dam line and Angus, Charolais, or University of Alberta hybrid bulls. Feed intake data were collected over a 5-yr period, with 2 groups (fall-winter and winter-spring) tested every year. Training and validation data sets were obtained by splitting the data into 2 distinct sets, by randomly splitting the data into training and testing sets based on sire family (split 1) in 5 replicates or by retaining all animals with no known pedigree relationships as the validation set (split 2). A total of 37,959 SNP were analyzed by single marker regression, of which only the top 100 that corresponded to a P-value <0.002 were retained. The 100 SNP were then analyzed using random regression BLUP, and only SNP that were jointly significant (P < 0.05) were included in the final marker panels. The marker effects from the selected panels were used to derive the molecular breeding values, which were calculated as a weighted sum of the number of copies of the more frequent allele at each SNP locus, with the weights being the allele substitution effects. The correlation between molecular breeding value and phenotype represented the accuracy of prediction. For all traits evaluated, accuracy across breeds was low, ranging between 0.007 and 0.414. Accuracy was least in data split 2, where the validation individuals had no pedigree relationship with animals in the training data. Given the low predictive ability observed, a large number of individuals may be needed for prediction when using such an admixed population. Further, these results suggest that breed composition of the target population in which the marker panels are likely to be used should be an important consideration when developing prediction equations across breeds, especially where an admixed population is used as the training data set. PMID- 21642495 TI - Phenotypic and genetic relationships among feeding behavior traits, feed intake, and residual feed intake in steers fed grower and finisher diets. AB - Data from a 3-yr feeding trial of crossbred steers (n = 331) were used to examine the relationship between feeding behavior traits and feed efficiency in steers fed grower and finisher diets, successively. There were 2 feeding periods each year whereby the steers were fed a grower diet in the first feeding period (P1) and a finisher diet in the second feeding period (P2). Each feeding period lasted for a minimum of 10 wk, ad libitum. In addition to feed intake, records on 3 measures of feeding behavior [feeding duration (FD), head-down time (HDT), and feeding frequency (FF)] were collected using the GrowSafe feeding system. Residual feed intake (RFI) was calculated by regression, after which the steers were classified as low (<0.5 SD), medium (+/-0.5 SD), or high (>0.5 SD) from the mean. The steers had greater (P < 0.001) FD, HDT, and FF when the grower diet was fed but greater feeding rate (FR) when the finisher diet was fed. Including the measures of feeding behavior as covariates to the feed intake prediction model containing ADG, metabolic midweight, and ultrasound backfat accounted for more variation in DMI than models that did not contain these additional parameters. The FD and HDT were significantly different (P < 0.05) among the RFI classes regardless of the feeding period, whereas no differences (P > 0.90) were found for FR among the RFI classes. For the growing period and finishing period, respectively, FD had phenotypic correlations with HDT (0.79, 0.83), FF (0.14, 0.55), DMI (0.38, 0.34), and FR (-0.34, -0.21). Heritability estimates in P1 and P2 for FD, HDT, and FF were 0.25 +/- 0.16, 0.14 +/- 0.11; 0.14 +/- 0.15, 0.09 +/- 0.10; and 0.56 +/- 0.19, 0.59 +/- 0.18, respectively. Genetic correlations between P1 and P2 were 0.91 +/- 0.26, 0.93 +/- 0.37, and 0.94 +/- 0.11 for FD, HDT, and FF, respectively. The results suggest that it may be appropriate to include feeding behavior traits as covariates to indicate measure(s) of animal activity in the calculation of RFI. Feeding behavior phenotypes were greater during the grower-fed period than the finisher-fed period. During these feeding periods, efficient steers exhibited fewer FF, shorter FD, and shorter HDT than inefficient steers. PMID- 21642496 TI - Genetic parameters for body weight, reproduction, and parasite resistance traits in the Creole goat. AB - We estimated the genetic parameters for BW, reproduction, and parasite resistance traits to implement a breeding program for the Creole goat. The traits were preweaning BW at 70 d of age (BW70d), BW at 11 mo of age (BW11), fecal egg count at 11 mo of age (FEC11) for all animals, packed cell volumes of lactating does (PCV), and their fertility (FER) and litter size (LS). We analyzed about 30 yr of data, which included 18,450 records on 11,970 animals from the INRA experimental flock in Guadeloupe (French West Indies). Heritability estimates were low for reproduction traits (0.11 +/- 0.02 for LS and FER) to moderate for production traits (0.32 +/- 0.03 for BW11; 0.20 +/- 0.03 and 0.08 +/- 0.02 for the direct and maternal heritability estimates of BW70d, respectively). Heritability estimates for gastrointestinal nematode resistance traits were situated in an intermediate range (0.13 +/- 0.05 for PCV and 0.18 +/- 0.04 for FEC11). Genetic correlations between FER, PCV, BW11, and the maternal effect of BW70d were altogether positive, whereas LS and FEC11 were almost uncorrelated phenotypically and genetically. These correlations are very favorable for setting up a breeding program, making it possible to improve BW, reproduction, and parasite resistance traits simultaneously. PMID- 21642497 TI - Effect of herbage depletion on short-term foraging dynamics and diet quality of steers grazing wheat pastures. AB - Two complementary experiments were completed to assess short-term foraging dynamics, diet quality, and ruminal degradation kinetics of herbage consumed by steers with 3 levels of herbage depletion. Experiment 1 was a behavioral study in which 2 ruminally cannulated steers were allocated to grazing scenarios simulating 3 levels of herbage depletion. These treatments included an ungrazed sward (control), as well as medium and high levels of herbage depletion. Grazing scenarios were sampled for sward surface height and amount of green leaf and stem before being grazed. Foraging dynamics were determined through measurements of bite rate, bite depth, eating step rate, eating distance, potential area consumed while grazing, and bites and intake per eating step. Also, quality of potential herbage consumed was estimated from hand-plucked herbage. In Exp. 2, ruminal degradation kinetics of DM for samples of herbage consumed (masticate) by steers during Exp. 1 were assessed in situ using 5 ruminally cannulated steers. The immediately soluble, degraded, and undegraded DM fractions were determined. The DM disappearance rate and lag times were determined from a nonlinear regression model, and the effective degradability of DM was calculated. Herbage depletion resulted in increased eating steps/minute, as well as the potential area harvested while grazing (P < 0.05) and reduced herbage intake/eating step (P < 0.05). Neither the herbage potentially consumed nor the ruminal degradation kinetics was affected by extent of depletion (P > 0.05). Under these experimental conditions, steers adapted their foraging dynamic and were able to sustain diet quality in the short term. These results imply that behavioral adaptations would make diet quality less sensitive to certain levels of herbage depletion. PMID- 21642498 TI - Growth performance of 20- to 50-kilogram pigs fed low-crude-protein diets supplemented with histidine, cystine, glycine, glutamic acid, or arginine. AB - The purpose of this investigation was to compare the growth performance of grower pigs fed low-CP, corn-soybean meal (C-SBM) AA-supplemented diets with that of pigs fed a positive control (PC) C-SBM diet with no supplemental Lys. Five experiments were conducted with Yorkshire crossbred pigs, blocked by BW (average initial and final BW were 21 and 41 kg, respectively) and assigned within block to treatment. Each treatment was replicated 4 to 6 times with 4 or 5 pigs per replicate pen. Each experiment lasted 28 d and plasma urea N was determined at the start and end of each experiment. All diets were formulated to contain 0.83% standardized ileal digestible Lys. All the experiments contained PC and negative control (NC) diets. The PC diet contained 18% CP and was supplemented with only DL-Met. The NC diet contained 13% CP and was supplemented with L-Lys, DL-Met, L Thr, and L-Trp. The NC + Ile + Val diet was supplemented with 0.10% Val + 0.06% Ile. The NC + Ile + Val diet was supplemented with either His (Exp. 1), Cys (Exp. 2), Gly (Exp. 2, 3, and 4), Glu (Exp. 3), Arg (Exp. 4), or combinations of Gly + Arg (Exp. 4 and 5) or Gly + Glu (Exp. 5). Treatment differences were considered significant at P < 0.10. In 3 of the 4 experiments that had PC and NC diets, pigs fed the NC diet had decreased ADG and G:F compared with pigs fed the PC diet. The supplementation of Ile + Val to the NC diet restored ADG in 4 out of 5 experiments. However, G:F was less than in pigs fed the PC diet in 1 experiment and was intermediate between the NC and PC diets in 3 experiments. Pigs fed supplemental Ile + Val + His had decreased G:F compared with pigs fed the PC. Pigs fed supplemental Cys to achieve 50:50 Met:Cys had decreased G:F compared with pigs fed the PC. Pigs fed Ile + Val + 0.224% supplemental Gly had similar ADG, greater ADFI, and decreased G:F compared with pigs fed the PC. Pigs fed Ile + Val + 0.52% supplemental Gly had ADG and G:F similar to that of pigs fed the PC. Pigs fed supplemental Glu had decreased G:F compared with pigs fed the PC. Pigs fed Ile + Val + 0.48% supplemental Arg had decreased G:F compared with pigs fed the PC. Pigs fed the diet supplemented with Gly + Arg had ADG and G:F similar to pigs fed the PC. Pigs fed the low-CP diets had reduced plasma urea N compared with pigs fed PC. The results of these experiments indicate that supplementing Gly or Gly + Arg to a low-CP C-SBM diet with 0.34% Lys, Met, Thr, Trp, Ile, and Val restores growth performance to be similar to that of pigs fed a PC diet with no Lys supplementation. PMID- 21642499 TI - Liver growth factor treatment restores cell-extracellular matrix balance in resistance arteries and improves left ventricular hypertrophy in SHR. AB - Liver growth factor (LGF) is an endogenous albumin-bilirubin complex with antihypertensive effects in spontaneously hypertensive rats (SHR). We assessed the actions of LGF treatment on SHR mesenteric resistance and intramyocardial arteries (MRA and IMA, respectively), heart, and vascular smooth muscle cells (VSMC). SHR and Wistar-Kyoto (WKY) rats treated with vehicle or LGF (4.5 MUg LGF/rat, 4 ip injections over 12 days) were used. Intra-arterial blood pressure was measured in anesthetized rats. The heart was weighted and paraffin-embedded. Proliferation, ploidy, and fibronectin deposition were studied in carotid artery derived VSMC by immunocytochemistry. In MRA, we assessed: 1) geometry and mechanics by pressure myography; 2) function by wire myography; 3) collagen by sirius red staining and polarized light microscopy, and 4) elastin, cell density, nitric oxide (NO), and superoxide anion by confocal microscopy. Heart sections were used to assess cell density and collagen content in IMA. Left ventricular hypertrophy (LVH) regression was assessed by echocardiography. LGF reduced blood pressure only in SHR. LGF in vitro or as treatment normalized the alterations in proliferation and fibronectin in SHR-derived VSMC with no effect on WKY cells. In MRA, LGF treatment normalized collagen, elastin, and VSMC content and passive mechanical properties. In addition, it improved NO availability through reduction of superoxide anion. In IMA, LGF treatment normalized perivascular collagen and VSMC density, improving the wall-to-lumen ratio. Paired experiments demonstrated a partial regression of SHR LVH by LGF treatment. The effective cardiovascular antifibrotic and regenerative actions of LGF support its potential in the treatment of hypertension and its complications. PMID- 21642500 TI - Cutaneous constitutive nitric oxide synthase activation in postural tachycardia syndrome with splanchnic hyperemia. AB - Models of microgravity are linked to excessive constitutive nitric oxide (NO) synthase (NOS), splanchnic vasodilation, and orthostatic intolerance. Normal-flow postural tachycardia syndrome (POTS) is a form of chronic orthostatic intolerance associated with splanchnic hyperemia. To test the hypothesis that there is excessive constitutive NOS in POTS, we determined whether cutaneous microvascular neuronal NO and endothelial NO are increased. We performed two sets of experiments in POTS and control subjects aged 21.4 +/- 2 yr. We used laser Doppler flowmetry to measure the cutaneous response to local heating as an indicator of bioavailable neuronal NO. To test for bioavailable endothelial NO, we infused intradermal acetylcholine through intradermal microdialysis catheters and used the selective neuronal NOS inhibitor l-N(omega)-nitroarginine-2,4-L diamino-butyric amide (N(omega), 10 mM), the selective inducible NOS inhibitor aminoguanidine (10 mM), the nonspecific NOS inhibitor nitro-l-arginine (NLA, 10 mM), or Ringer solution. The acetylcholine dose response and the NO-dependent plateau of the local heating response were increased in POTS compared with those in control subjects. The local heating plateau was significantly higher, 98 +/- 1%maximum cutaneous vascular conductance (%CVC(max)) in POTS compared with 88 +/- 2%CVC(max) in control subjects but decreased to the same level with N(omega) (46 +/- 5%CVC(max) in POTS compared with 49 +/- 4%CVC(max) in control) or with NLA (45 +/- 3%CVC(max) in POTS compared with 47 +/- 4%CVC(max) in control). Only NLA blunted the acetylcholine dose response, indicating that NO produced by endothelial NOS was released by acetylcholine. Aminoguanidine was without effect. This is consistent with increased endothelial and neuronal NOS activity in normal flow POTS. PMID- 21642501 TI - Inhalation of hydrogen gas attenuates left ventricular remodeling induced by intermittent hypoxia in mice. AB - Sleep apnea syndrome increases the risk of cardiovascular morbidity and mortality. We previously reported that intermittent hypoxia increases superoxide production in a manner dependent on nicotinamide adenine dinucleotide phosphate and accelerates adverse left ventricular (LV) remodeling. Recent studies have suggested that hydrogen (H(2)) may have an antioxidant effect by reducing hydroxyl radicals. In this study, we investigated the effects of H(2) gas inhalation on lipid metabolism and LV remodeling induced by intermittent hypoxia in mice. Male C57BL/6J mice (n = 62) were exposed to intermittent hypoxia (repetitive cycle of 1-min periods of 5 and 21% oxygen for 8 h during daytime) for 7 days. H(2) gas (1.3 vol/100 vol) was given either at the time of reoxygenation, during hypoxic conditions, or throughout the experimental period. Mice kept under normoxic conditions served as controls (n = 13). Intermittent hypoxia significantly increased plasma levels of low- and very low-density cholesterol and the amount of 4-hydroxy-2-nonenal-modified protein adducts in the LV myocardium. It also upregulated mRNA expression of tissue necrosis factor alpha, interleukin-6, and brain natriuretic peptide, increased production of superoxide, and induced cardiomyocyte hypertrophy, nuclear deformity, mitochondrial degeneration, and interstitial fibrosis. H(2) gas inhalation significantly suppressed these changes induced by intermittent hypoxia. In particular, H(2) gas inhaled at the timing of reoxygenation or throughout the experiment was effective in preventing dyslipidemia and suppressing superoxide production in the LV myocardium. These results suggest that inhalation of H(2) gas was effective for reducing oxidative stress and preventing LV remodeling induced by intermittent hypoxia relevant to sleep apnea. PMID- 21642502 TI - Heterogenous vasodilator pathways underlie flow-mediated dilation in men and women. AB - This study investigated the sex differences in the contribution of nitric oxide (NO) and prostaglandins (PGs) to flow-mediated dilation (FMD). Radial artery (RA) FMD, assessed as the dilatory response to 5-min distal cuff occlusion, was repeated after three separate brachial artery infusions of saline (SAL), N(G) monomethyl-L-arginine (L-NMMA), and ketorolac (KETO) + L-NMMA in healthy younger men (M; n = 8) and women (W; n = 8). In eight subjects (4 M, 4W) RA FMD was reassessed on a separate day with drug order reversed (SAL, KETO, and L-NMMA + KETO). RA FMD was calculated as the peak dilatory response observed relative to baseline (%FMD) and expressed relative to the corresponding area under the curve shear stress (%FMD/AUC SS). L-NMMA reduced %FMD similarly and modestly (P = 0.68 for sex * trial interaction) in M and W (all subjects: 10.0 +/- 3.8 to 7.6 +/- 4.7%; P = 0.03) with no further effect of KETO (P = 0.68). However, all sex * trial and trial effects on %FMD/AUC SS for l-NMMA and KETO + l-NMMA were insignificant (all P > 0.20). There was also substantial heterogeneity of the magnitude and direction of dilator responses to blockade. After l-NMMA infusion, subjects exhibited both reduced (n = 14; range: 11 to 78% decrease) and augmented (n = 2; range: 1 to 96% increase) %FMD. Following KETO + l-NMMA, seven subjects exhibited reduced dilation (range: 10 to 115% decrease) and nine subjects exhibited augmented dilation (range: 1 to 212% increase). Reversing drug order did not change the nature of the findings. These findings suggest that RA FMD is not fully or uniformly NO dependent in either men or women, and that there is heterogeneity in the pathways underlying the conduit dilatory response to ischemia. PMID- 21642503 TI - Glucan phosphate attenuates myocardial HMGB1 translocation in severe sepsis through inhibiting NF-kappaB activation. AB - Myocardial dysfunction is a major consequence of septic shock and contributes to the high mortality of sepsis. High-mobility group box 1 (HMGB1) serves as a late mediator of lethality in sepsis. We have reported that glucan phosphate (GP) attenuates cardiac dysfunction and increases survival in cecal ligation and puncture (CLP)-induced septic mice. In the present study, we examined the effect of GP on HMGB1 translocation from the nucleus to the cytoplasm in the myocardium of septic mice. GP was administered to mice 1 h before induction of CLP. Sham operated mice served as control. The levels of HMGB1, Toll-like receptor 4 (TLR4), and NF-kappaB binding activity were examined. In an in vitro study, H9C2 cardiomyoblasts were treated with lipopolysaccharide (LPS) in the presence or absence of GP. H9C2 cells were also transfected with Ad5-IkappaBalpha mutant, a super repressor of NF-kappaB activity, before LPS stimulation. CLP significantly increased the levels of HMGB1, TLR4, and NF-kappaB binding activity in the myocardium. In contrast, GP administration attenuated CLP-induced HMGB1 translocation from the nucleus to the cytoplasm and reduced CLP-induced increases in TLR4 and NF-kappaB activity in the myocardium. In vitro studies showed that GP prevented LPS-induced HMGB1 translocation and NF-kappaB binding activity. Blocking NF-kappaB binding activity by Ad5-IkappaBalpha attenuated LPS-induced HMGB1 translocation. GP administration also reduced the LPS-stimulated interaction of HMGB1 with TLR4. These data suggest that attenuation of HMGB1 translocation by GP is mediated through inhibition of NF-kappaB activation in CLP induced sepsis and that activation of NF-kappaB is required for HMGB1 translocation. PMID- 21642504 TI - Angiotensin II-induced process of angiogenesis is mediated by spleen tyrosine kinase via VEGF receptor-1 phosphorylation. AB - Spleen tyrosine kinase (Syk), expressed in endothelial cells, has been implicated in migration and proliferation and in vasculogenesis. This study was conducted to determine the contribution of Syk and the underlying mechanism to the angiogenic effect of ANG II and VEGF. Angiogenesis was determined by tube formation from the endothelial cell line EA.hy926 (EA) and human umbilical vein endothelial cells (HUVECs) and microvessel sprouting in rat aortic rings. ANG II (10 nM), EGF (30 ng/ml), and VEGF (50 ng/ml) stimulated EA cells and HUVECs to form tubular networks and increased aortic sprouting; these effects were blocked by VEGF receptor-1 and Flt-1 antibody (Flt-1/Fc) but not by the VEGF receptor-2 (Flk-1) antagonist SU-1498. ANG II increased the phosphorylation of Flt-1 but not Flk-1, whereas VEGF increased the phosphorylation of both receptors in EA cells and HUVECs. VEGF expression elicited by ANG II was not altered by Flt-1/Fc or SU 1498. EGF stimulated tube formation from EA cells and HUVECs and Flt-1 phosphorylation and aortic sprouting, which were blocked by the EGF receptor antagonist AG-1478 and Flt-1/Fc but not by SU-1498. ANG II-, EGF-, and VEGF induced tube formation and aortic sprouting were attenuated by the Syk inhibitor piceatannol and by Syk short hairpin interfering (sh)RNA and small interfering RNA, respectively. ANG II, EGF, and VEGF increased Syk phosphorylation, which was inhibited by piceatannol and Syk shRNA in EA cells and HUVECs. Neither piceatannol nor Syk shRNA altered ANG II-, EGF-, or VEGF-induced phosphorylation of Flt-1. These data suggest that ANG II stimulates angiogenesis via transactivation of the EGF receptor, which promotes the phosphorylation of Flt-1 and activation of Syk independent of VEGF expression. PMID- 21642505 TI - Contribution of nerve growth factor to upregulation of P2X3 expression in DRG neurons of rats with femoral artery occlusion. AB - Femoral artery occlusion augments the sympathetic nerve and pressor responses to muscle contraction and muscle metabolites injected into the arterial blood supply of the hindlimb muscles in rats. The underlying mechanism by which these reflex responses are enhanced after muscle vascular insufficiency is unclear. Purinergic P2X(3) receptor has been reported to contribute to the metabolic component of the exercise pressor reflex. Thus the purpose of this study was to examine if chronic femoral occlusion would alter the expression of P2X(3) in dorsal root ganglion (DRG) neurons of rats. Also, P2X(3)-mediated sympathetic responsiveness was examined after femoral occlusion. In addition, the role played by nerve growth factor (NGF) in regulating the expression and response of P2X(3) was examined. Western blot analysis showed that 24 h of femoral ligation increased the levels of P2X(3) (optical density: 0.93 +/- 0.07 in control and 1.37 +/- 0.10 after occlusion; P < 0.05 vs. control). The fluorescence immunohistochemistry further demonstrated that the occlusion elevated P2X(3) expression in DRG neurons (percentage of P2X(3)-positive cells: 33 +/- 3% in control and 51 +/- 3% in occlusion; P < 0.05 vs. control). Furthermore, the results showed that responses of renal sympathetic nerve activity and blood pressure to stimulation of P2X were greater in occluded rats than responses in control rats by injection of alpha,beta-methylene ATP into the arterial blood supply of the hindlimb muscle. Finally, infusion of NGF in the hindlimb muscles of healthy rats increased P2X(3) (optical density: 0.98 +/- 0.12 in control and 1.37 +/- 0.16 with NGF; P < 0.05 vs. control). The pressor response to injection of alpha,beta-methylene ATP was increased in the rats with NGF infusion. Likewise, blocking NGF attenuated exaggeration of the reflex response induced by alpha,beta-methylene ATP in occluded rats. The findings of this study suggest that the levels of P2X(3) in primary afferent neurons are upregulated as the blood supply to the hindlimb is deficient under ischemic conditions, leading to augmentation of the muscle reflex. NGF is closely related to increases in P2X(3) receptor expression and response. PMID- 21642506 TI - A Pmk1-interacting gene is involved in appressorium differentiation and plant infection in Magnaporthe oryzae. AB - In the rice blast fungus Magnaporthe oryzae, the PMK1 mitogen-activated protein (MAP) kinase gene regulates appressorium formation and infectious growth. Its homologs in many other fungi also play critical roles in fungal development and pathogenicity. However, the targets of this important MAP kinase and its interacting genes are not well characterized. In this study, we constructed two yeast two-hybrid libraries of M. oryzae and screened for Pmk1-interacting proteins. Among the nine Pmk1-interacting clones (PICs) identified, two of them, PIC1 and PIC5, were selected for further characterization. Pic1 has one putative nuclear localization signal and one putative MAP kinase phosphorylation site. Pic5 contains one transmembrane domain and two functionally unknown CTNS (cystinosin/ERS1p repeat) motifs. The interaction of Pmk1 with Pic1 or Pic5 was confirmed by coimmunoprecipitation assays. Targeted gene deletion of PIC1 had no apparent effects on vegetative growth and pathogenicity but resulted in a significant reduction in conidiation and abnormal germ tube differentiation on onion epidermal cells. Deletion of PIC5 led to a reduction in conidiation and hyphal growth. Autolysis of aerial hyphae became visible in cultures older than 4 days. The pic5 mutant was defective in germ tube growth and appressorium differentiation. It was reduced in appressorial penetration and virulence on the plant. Both PIC1 and PIC5 are conserved in filamentous ascomycetes, but none of their orthologs have been functionally characterized. Our data indicate that PIC5 is a novel virulence factor involved in appressorium differentiation and pathogenesis in M. oryzae. PMID- 21642507 TI - Rab11 function in Trypanosoma brucei: identification of conserved and novel interaction partners. AB - The Ras-like GTPase Rab11 is implicated in multiple aspects of intracellular transport, including maintenance of plasma membrane composition and cytokinesis. In metazoans, these functions are mediated in part via coiled-coil Rab11 interacting proteins (FIPs) acting as Rab11 effectors. Additional interaction between Rab11 and the exocyst subunit Sec15 connects Rab11 with exocytosis. We find that FIPs are metazoan specific, suggesting that other factors mediate Rab11 functions in nonmetazoans. We examined Rab11 interactions in Trypanosoma brucei, where endocytosis is well studied and the role of Rab11 in recycling well documented. TbSec15 and TbRab11 interact, demonstrating evolutionary conservation. By yeast two-hybrid screening, we identified additional Rab11 interaction partners. Tb927.5.1640 (designated RBP74) interacted with both Rab11 and Rab5. RBP74 shares a coiled-coil architecture with metazoan FIPs but is unrelated by sequence and appears to play a role in coordinating endocytosis and recycling. A second coiled-coil protein, Tb09.211.4830 (TbAZI1), orthologous to AZI1 in Homo sapiens, interacts exclusively with Rab11. AZI1 is restricted to taxa with motile cilia/flagella. These data suggest that Rab11 functions are mediated by evolutionarily conserved (i.e., AZI1 and Sec15) and potentially lineage-specific (RBP74) interactions essential for the integration of the endomembrane system. PMID- 21642508 TI - Modulation of morphogenesis in Candida albicans by various small molecules. AB - The pathogenic yeast Candida albicans, a member of the mucosal microbiota, is responsible for a large spectrum of infections, ranging from benign thrush and vulvovaginitis in both healthy and immunocompromised individuals to severe, life threatening infections in immunocompromised patients. A striking feature of C. albicans is its ability to grow as budding yeast and as filamentous forms, including hyphae and pseudohyphae. The yeast-to-hypha transition contributes to the overall virulence of C. albicans and may even constitute a target for the development of antifungal drugs. Indeed, impairing morphogenesis in C. albicans has been shown to be a means to treat candidiasis. Additionally, a large number of small molecules such as farnesol, fatty acids, rapamycin, geldanamycin, histone deacetylase inhibitors, and cell cycle inhibitors have been reported to modulate the yeast-to-hypha transition in C. albicans. In this minireview, we take a look at molecules that modulate morphogenesis in this pathogenic yeast. When possible, we address experimental findings regarding their mechanisms of action and their therapeutic potential. We discuss whether or not modulating morphogenesis constitutes a strategy to treat Candida infections. PMID- 21642509 TI - Multiple roles of Ypd1 phosphotransfer protein in viability, stress response, and virulence factor regulation in Cryptococcus neoformans. AB - Ypd1 is a key phosphorelay protein that controls eukaryotic two-component systems, but its function in Cryptococcus neoformans is not known. Here, we report that Ypd1 is required for the viability of C. neoformans via the Hog1 mitogen-activated protein kinase (MAPK) pathway but plays multiple cellular roles in both a Hog1-dependent and -independent manner. PMID- 21642510 TI - Clustering of nuclei in multinucleated hyphae is prevented by dynein-driven bidirectional nuclear movements and microtubule growth control in Ashbya gossypii. AB - During filamentous fungus development, multinucleated hyphae employ a system for long-range nuclear migration to maintain an equal nuclear density. A decade ago the microtubule motor dynein was shown to play a central role in this process. Previous studies with Ashbya gossypii revealed extensive bidirectional movements and bypassings of nuclei, an autonomous cytoplasmic microtubule (cMT) cytoskeleton emanating from each nucleus, and pulling of nuclei by sliding of cMTs along the cortex. Here, we show that dynein is the sole motor for bidirectional movements and bypassing because these movements are concomitantly decreased in mutants carrying truncations of the dynein heavy-chain DYN1 promoter. The dynactin component Jnm1, the accessory proteins Dyn2 and Ndl1, and the potential dynein cortical anchor Num1 are also involved in the dynamic distribution of nuclei. In their absence, nuclei aggregate to different degrees, whereby the mutants with dense nuclear clusters grow extremely long cMTs. As in budding yeast, we found that dynein is delivered to cMT plus ends, and its activity or processivity is probably controlled by dynactin and Num1. Together with its role in powering nuclear movements, we propose that dynein also plays (directly or indirectly) a role in the control of cMT length. Those combined dynein actions prevent nuclear clustering in A. gossypii and thus reveal a novel cellular role for dynein. PMID- 21642511 TI - Pulsed shortwave treatment in women with knee osteoarthritis: a multicenter, randomized, placebo-controlled clinical trial. AB - BACKGROUND: Several forms of conservative treatment have been the focus of many recent studies in knee osteoarthritis (OA). Among these techniques, the application of pulsed shortwave (PSW) treatment has been widely used, but the optimal dose and application time have not been well established. Objective The purposes of this study were: (1) evaluate the effect of PSW treatment in different doses and (2) to compare low-dose and high-dose PSW groups with control and placebo groups. Design This was a randomized clinical trial. Setting The study was conducted in the physical therapy department of 2 large urban hospitals. Patients One hundred twenty-one women (mean age=60 years, SD=9) with a diagnosis of knee OA participated in the study. INTERVENTION AND MEASUREMENTS: Participants were distributed randomly into 4 groups: 35 participants did not receive any treatment (control group), 23 received a placebo treatment, 32 received low-dose PSW treatment (power of 14.5 W, treatment duration of 19 minutes, and total energy of 17 kJ), and 31 received high-dose PSW treatment (power of 14.5 W, treatment duration of 38 minutes, and total energy of 33 kJ). An 11-point numerical pain rating scale and the Knee Osteoarthritis Outcome Score were used to assess pain and function in 3 stages: at initial evaluation (pretreatment), immediately after treatment, and at 12-month follow-up. RESULTS: The 4 groups were homogeneous prior to treatment with respect to demographics, pain, and functional scale data. The results demonstrated the short-term effectiveness of the PSW at low and high doses in patients with knee OA. Both treatment groups showed a significant reduction in pain and improvement in function compared with the control and placebo groups (effect size: range=20.0 23.4 for the low-dose PSW group and range=15.7-16.5 for the high-dose PSW group). There were no differences in results between PSW doses, although a low dose of PSW appeared to be more effective in the long term. Limitations These results were achieved without physical exercise, which could have positively influenced the results. CONCLUSIONS: Pulsed shortwave treatment is an effective method for pain relief and improvement of function and quality of life in the short term in women with knee OA. On the basis of the results, application of PSW treatment is recommended in the female population with knee OA. However, conclusions regarding the 12-month follow-up should be analyzed carefully due to the high dropout rate. PMID- 21642512 TI - Patient stories about their dialysis experience biases others' choices regardless of doctor's advice: an experimental study. AB - BACKGROUND: Renal services provide resources to support patients in making informed choices about their dialysis modality. Many encourage new patients to talk with those already experiencing dialysis. It is unclear if these stories help or hinder patients' decisions, and few studies have been conducted into their effects. We present two studies comparing the impact of patient and doctor stories on hypothetical dialysis modality choices among an experimental population. METHODS: In total, 1694 participants viewed online information about haemodialysis and continuous cycling peritoneal dialysis and completed a questionnaire. In Study 1, using actors, treatment information was varied by presenter (Doctor, Patient), order of presenter (Patient first, Doctor first) and mode of delivery (written, video). Information in Study 2 was varied (using actors) by presenter (Doctor, Patient), order of presenter (Patient first, Doctor first), inclusion of a decision table (no table, before story, after story) and sex of the 'patient' (male, female) and 'Doctor' (male, female). Information was controlled to ensure comparable content and comprehensibility. RESULTS: In both studies, participants were more likely to choose the dialysis modality presented by the patient rather than that presented by the doctor. There was no effect for mode of delivery (video versus written) or inclusion of a decision table. CONCLUSIONS: As 'new' patients were making choices based on past patient experience of those already on dialysis, we recommend caution to services using patient stories about dialysis to support those new to the dialysis in delivering support to those who are new to the decision making process for dialysis modality. PMID- 21642513 TI - Quantification of glomerular number and size distribution in normal rat kidneys using magnetic resonance imaging. AB - BACKGROUND: Glomerular number and size are important risk factors for chronic kidney disease (CKD) and cardiovascular disease and have traditionally been estimated using invasive techniques. Here, we report a novel technique to count and size every glomerulus in the rat kidney using magnetic resonance imaging (MRI). METHODS: The ferromagnetic nature of cationized ferritin allowed visualization of single glomeruli in high-resolution susceptibility-weighted MRI. A segmentation algorithm was used to identify and count all glomeruli within the whole kidney. To prove our concept, we estimated total glomerular number and mean glomerular volume of each kidney using design-based stereology. RESULTS: The glomerular counts obtained with MRI agreed well with estimates obtained using traditional methods [MRI, 32 785 (3117); stereology, 35 132 (3123)]. For the first time, the glomerular volume distribution for the entire kidney is shown. Additionally, the method is substantially faster than the current methods. CONCLUSIONS: MRI provides a new method for measuring these important microanatomical markers of disease risk and leads the way to in vivo analysis of these parameters, including longitudinal studies of animal models of CKD. PMID- 21642514 TI - Phase II clinical trial of neoadjuvant trabectedin in patients with advanced localized myxoid liposarcoma. AB - BACKGROUND: To evaluate neoadjuvant trabectedin (1.5 mg/m(2) 24-h i.v. infusion every 3 weeks; three to six cycles) in patients with locally advanced myoxid liposarcoma (ML) previously untreated with chemotherapy or radiation. PATIENTS AND METHODS: Primary efficacy end point was pathological complete response (pCR) or tumoral regression rate. Objective response according to RECIST (v.1.0) was a secondary end point. RESULTS: Three of 23 assessable patients had pCR [13%; 95% confidence interval (CI), 3% to 34%]. Furthermore, very good and moderate histological responses were observed in another 2 and 10 patients, respectively. Histological decrement in the cellular and vascular tumor component and maturation of tumor cells to lipoblasts were observed in both myoxid and myoxid/round cell variants. Seven patients had partial response according to RECIST (objective response rate of 24%; 95% CI, 10% to 44%). No disease progression was reported. Neoadjuvant trabectedin was usually well tolerated, with a safety profile similar to that described in patients with soft tissue sarcoma or other tumor types. CONCLUSION: Trabectedin 1.5 mg/m(2) given as a 24-h i.v. infusion every 3 weeks is a therapeutic option in the neoadjuvant setting of ML. PMID- 21642515 TI - High independent prognostic and predictive value of circulating tumor cells compared with serum tumor markers in a large prospective trial in first-line chemotherapy for metastatic breast cancer patients. AB - BACKGROUND: Circulating tumor cells (CTCs) are a prognostic marker in metastatic breast cancer, but comparisons with serum tumor markers (CA 15-3, carcinoembryonic antigen and lactate dehydrogenase) variations are needed. PATIENTS AND METHODS: CTCs were counted with CellSearch(r) at baseline, before cycle 2 (C2) and cycle 3 or 4 (C3/4) in 267 metastatic breast cancer patients on first-line chemotherapy with/without targeted therapy. RESULTS: Baseline CTC detection rate was 65% with >=1 CTC/7.5 ml threshold and 44% with >=5 CTC/7.5 ml and was independent of subtypes (luminal, triple negative, human epithelial growth factor receptor 2 (HER2)+). CTCs were associated with tumor markers, bone/liver involvement, tumor burden and performance status. CTC detection >=1 CTC/7.5 ml was a strong prognostic factor for progression-free survival (PFS), P < 0.0001. Threshold of CTC >=5 was statistically significant for PFS and overall survival (OS), P = 0.03 on multivariate analysis. Among patients with >=5 CTC/7.5 ml at baseline, 50% had <5 CTC/7.5 ml at C2. Changes were correlated with both PFS and OS (P < 0.0001). All patients receiving anti-HER2 therapy had <5 CTC/7.5 ml after three cycles of treatment. CONCLUSION: This is the largest prospective series validating the prognostic value of CTC independently from serum tumor marker. Elevated CTCs before C2 are an early predictive marker of poor PFS and OS, which could be used to monitor treatment benefit. CTC decrease under treatment seems stronger with targeted therapy. PMID- 21642516 TI - Evaluating postdoctoral dental candidates: assessing the need and recommendations for a national qualifying examination. AB - Increasingly, U.S. dental schools report pass/fail grades and do not rank students. In addition, the Joint Commission on National Dental Examinations will report National Board Dental Examination (NBDE) scores as pass/fail after January 1, 2012. This article discusses how these changes will force postdoctoral dental program directors to modify how they assess candidates and how noncognitive evaluations might enhance those assessments. The authors propose developing a national qualifying examination for postdoctoral dental programs that will measure knowledge, decision making, and noncognitive traits including empathy, self-confidence, integrity, and emotional intelligence. Without NBDE scores, class rank, and GPA as a basis for decision making, a single national qualifying examination would assist postdoctoral programs in selecting high-quality candidates based on knowledge, critical thinking skills, and noncognitive traits. PMID- 21642517 TI - Perceptions of predoctoral dental education and practice patterns in special care dentistry. AB - The objective of this research project was to compare alumni perceptions of predoctoral dental education in the care and management of patients with complex needs to alumni practice patterns. Alumni from the University of the Pacific Arthur A. Dugoni School of Dentistry who graduated from 1997 to 2007 were surveyed regarding perceptions of their predoctoral education in the care of patients categorized and defined as medically compromised, frail elders, and developmentally disabled, as well as their practice patterns. Perceptions were rated on a Likert scale. Regression analyses were utilized. Three primary relationships were identified: 1) positive relationships emerged between perceptions of educational value, as students and practitioners, of the training they received compared to percentages of medically compromised patients they currently treat (p<=0.05); 2) after practice experience, 2003-07 graduates reported significantly higher value of their education in this area compared to 1997-2002 graduates; and 3) alumni who reported treating more patients with complex needs during school reported treating significantly more of these patients in practice (p<=0.05). We conclude that alumni who reported educational experiences as more valuable treat more patients with complex needs compared to those who valued them less. Alumni who reported having more opportunities to treat patients with complex needs as students treat a higher percentage of those patients than those reporting fewer. Even positive perceptions may underestimate the value of educational experiences as they relate to future practice. PMID- 21642518 TI - Teledentistry-assisted, affiliated practice for dental hygienists: an innovative oral health workforce model. AB - The 2010 U.S. Patient Protection and Affordable Care Act (PPACA) calls for training programs to develop mid-level dental health care providers to work in areas with underserved populations. In 2004, legislation was passed in Arizona allowing qualified dental hygienists to enter into an affiliated practice relationship with a dentist to provide oral health care services for underserved populations without general or direct supervision in public health settings. In response, the Northern Arizona University (NAU) Dental Hygiene Department developed a teledentistry-assisted, affiliated practice dental hygiene model that places a dental hygienist in the role of the mid-level practitioner as part of a digitally linked oral health care team. Utilizing current technologies, affiliated practice dental hygienists can digitally acquire and transmit diagnostic data to a distant dentist for triage, diagnosis, and patient referral in addition to providing preventive services permitted within the dental hygiene scope of practice. This article provides information about the PPACA and the Arizona affiliated practice dental hygiene model, defines teledentistry, identifies the digital equipment used in NAU's teledentistry model, give an overview of NAU's teledentistry training, describes NAU's first teledentistry clinical experience, presents statistical analyses and evaluation of NAU students' ability to acquire diagnostically efficacious digital data from remote locations, and summarizes details of remote applications of teledentistry assisted, affiliated practice dental hygiene workforce model successes. PMID- 21642519 TI - Situational judgment tests as a new tool for dental student selection. AB - Cognitive tests are used to select students into dental school, yet cognitive predictors explain only part of the variance in academic performance. Therefore, interviews and personality tests are often used to measure noncognitive (e.g., interpersonal) characteristics. Recently, situational judgment tests (SJTs) have drawn attention since there is evidence that SJTs can be valid predictors in medical admission contexts. This study examines the validity of an SJT measuring interpersonal skills for predicting academic performance of dental students. Incremental validity over cognitive tests is also examined. In this study, 796 dental students who passed the admission exam for medical and dental students in Flanders, Belgium, and enrolled in one of the two Flemish dental schools were evaluated. Grade point average (GPA) in the five years of dental studies served as the criterion. Corrected correlation between the cognitive tests of the admission exam and GPA equaled .38. Their validity dropped from .45 (year 1) to .18 (year 5). However, the validity of the SJT increased from .05 (year 1) to .20 (year 5). The SJT had incremental validity in year 5. Dental admissions committees that envision assessing a broad set of capabilities might consider using an SJT as a valuable supplement to cognitive tests. Future research needs to confirm these findings with job performance as another criterion. PMID- 21642520 TI - Dental student perceptions of predoctoral implant education and plans for providing implant treatment. AB - This study aims to identify dental students' perceptions of pre-patient care laboratory exercises (PCLEs) and clinical experiences that influence their future plans for providing implant care. One of two questionnaires was administered to dental student classes at one dental school (D2: Survey 1; D3 and D4: Survey 2). Future plans as graduates to provide implant diagnosis and treatment planning (DxTP), restoration of single-tooth implants (STIs), and implant-retained overdentures (IODs) were cross-sectionally assessed along with potential influences such as PCLE, clinical experiences, gender, and class. The majority of students planned to provide implant services after graduation (DxTP 68.9 percent; STI 61.2 percent; IOD 62.1 percent). Bivariately, males reflected more preparedness from PCLEs than females (p=.002) and the D2 students more than D3 and D4 students (p<.001). Multivariate models revealed the perceived preparedness from PCLEs generally had the strongest association with future plans for performing implant therapy. However, this varied by gender and class. These findings indicate that PCLEs are important for their influence on students' future plans to provide implant therapy. However, further studies are needed to validate actual PCLEs and clinical implant practices (both longitudinally and for other schools) and to determine educational interventions to optimize the provision of implant care. PMID- 21642521 TI - Physical fitness course in the dental curriculum and prevention of low back pain. AB - Physical and psychosocial stressors in dental schools are associated with adverse health outcome, including low back pain. The aim of this study was to evaluate the physical fitness course included in a dental school's curriculum with regard to prevention of low back pain. Ninety first-year and sixty-two final-year dental students completed an anonymous questionnaire on physical fitness habits and low back pain. Fifty voluntarily participated in the Astrand bicycle ergometer test. The questionnaire revealed that 37 percent of the students have weekly physical exercise only during the physical fitness course included in the curriculum and 62.5 percent of the students reported low back pain. Final-year dental students had significantly more low back pain than first-year dental students (r=0.21, chi(2)=7.91, p=0.005). Female students had significantly more low back pain than male students (r=0.28, chi(2)=6.61, p=0.0101). The Astrand test revealed that students who attended the physical fitness course had significantly better physical fitness (p=0.008) than those who did not. Students who exercised more regularly had significantly less low back pain (r =-0.19, chi(2)=11.89, p<0.01) than those who did not. We conclude that participation in a physical fitness course leads to improved low back health for dental students and may prevent low back pain among final-year dental students. PMID- 21642522 TI - Application data as an indicator for post-Katrina recovery of LSU Postdoctoral dental programs. AB - Devastated by the aftermath of Hurricane Katrina in August 2005, the New Orleans campus of the Louisiana State University School of Dentistry was closed for two years. With help from the university's Systems Office, the school created a temporary campus in Baton Rouge, seventy-five miles from the New Orleans campus. The eight postdoctoral education programs, however, were forced to find clinic and seminar space in other facilities and cities. Of the seventy-nine students in these programs, only sixteen did not continue after the storm. This article describes how each program maintained its curriculum while the main campus was closed. By comparing numbers of student applications to these programs in each year since Hurricane Katrina with the average baseline for each in the ten years preceding the storm, this article illustrates the current viability of these residency programs. Total applications in 2005-07 were significantly lower than baseline measures, but applications in 2008 and 2009 have returned to pre-storm levels. A comparison of these application numbers with national trends also demonstrates that these programs have rebounded from the effects of Hurricane Katrina. PMID- 21642523 TI - Effectiveness of web-based teaching modules: test-enhanced learning in dental education. AB - The purpose of our study was to evaluate the effectiveness of self-tests as a component of web-based self-instruction in predoctoral orthodontics and pediatric dentistry. To this end, the usage patterns of online teaching modules and self tests by students enrolled in three courses at the University of North Carolina at Chapel Hill School of Dentistry were monitored and correlated to final exam grade and course average. We recorded the frequency of access to thirty relevant teaching modules and twenty-nine relevant self-tests for 157 second- and third year D.D.S. students during the course of our data collection. There was a statistically significant positive correlation between frequency of accessing self-tests and course performance in one course that was totally based on self instruction with seminars and multiple-choice examination (Level IV): Spearman correlation between frequency of self-test access and final exam grade, rho=0.23, p=0.044; correlation between frequency of self-test access and course average: rho=0.39, p=0.0004. In the other two courses we monitored, which included content beyond self-instruction with self-tests, the correlations were positive but not statistically significant. The students' use of online learning resources varied significantly from one course (Level I) to the next (Level II): Wilcoxon matched pairs signed-rank tests, S=-515.5, p=.0057 and S=1086, p<0.0001. The data from this study suggest that increased use of web-based self-tests may be correlated with more effective learning in predoctoral dental education by virtue of the testing effect and that dental students' usage of resources for learning changes significantly over the course of their education. PMID- 21642524 TI - A medical crisis management simulation activity for pediatric dental residents and assistants. AB - Dentists are expected to deliver safe and pain-free dental procedures after they graduate from dental school. This includes using local anesthetics and sedative drugs that may be associated with side effects and complications that can lead to crisis situations. This study postulated that teaching medical crisis management to dental residents and assistants using human patient simulation (HPS) would improve their confidence in managing crisis situations in the real world. Four medical crisis scenarios were designed and programmed into a pediatric simulator. The scenarios included anaphylaxis, laryngospasm during procedural sedation, sedative medication overdose, and multiple drug interaction with cardiac arrhythmia. The simulation room was outfitted with an authentic dental operatory and emergency equipment to enhance the realism. One first-or second-year pediatric dentistry resident and a staff dental assistant were assigned as a team to participate in each ten-minute scenario followed by a debriefing session. At the end of the sessions, the participants completed an anonymous survey regarding the simulation experience. There were a total of twenty-four participants, 91.7 percent of whom felt that HPS was a good tool for learning medical crisis and that they will be more confident in managing a similar situation in the dental office after this experience. A majority of the participants felt that using HPS as a tool to teach crisis management is an acceptable and valuable technique to help improve their confidence in managing crisis situations that may occur in dental offices. PMID- 21642525 TI - Preliminary development and evaluation of online tobacco and alcohol modules for dental students. AB - Tobacco use and heavy alcohol consumption are major risk factors for the development of oral and pharyngeal cancer (OPC). Detection and modification of these risks by dentists are keys in preventing OPC. While dentists are encouraged to screen patients for tobacco and alcohol use and educate them about the oral health risks they pose, dental students receive little formalized training in this area. This pilot project was designed to develop and evaluate two online training modules for dental students: one on tobacco and oral health risk factors, and one on methods of alcohol screening. Results indicated that online tobacco/alcohol education for dental students is feasible. The modules resulted in meaningful improvement in dental students' knowledge of tobacco and alcohol use as well as alcohol screening methods. The alcohol module resulted in statistically significant increases in intention to screen patients for alcohol use and in comfort level in performing alcohol screening. PMID- 21642526 TI - Teaching clinically relevant dental anatomy in the dental curriculum: description and assessment of an innovative module. AB - The primary objective of the preclinical dental anatomy course in the predoctoral dental curriculum is to introduce students to cognitive and psychomotor skills related to the morphology and spatial and functional relationships of human dentition. Traditionally, didactic content for the subject is found in textbooks and course manuals and summarized by the faculty in lectures to the entire class. Psychomotor skills associated with recognition and reproduction of tooth morphology are traditionally learned by examining preserved tooth specimens and their cross-sections, combined with producing two-dimensional line drawings and carving teeth from wax blocks. These activities have little direct clinical application. In most cases, students are passive in the learning process, and assessment of student performance is unilateral and subjective. A recently revised dental anatomy module at the University of Illinois at Chicago College of Dentistry integrates independent class preparation with active small-group discussion and patient scenario-based wax-up exercises to replace missing tooth structure on manikin teeth. The goal of the revision is to shift emphasis away from decontextualized technical learning toward more active and clinically applicable learning that improves conceptual understanding while contributing to early acquisition of psychomotor skills. This article describes the rationale, components, and advantages of the revised module and presents a pre-post comparison of student learning outcomes for three class cohorts (N=203). PMID- 21642527 TI - Student tutors for problem-based learning in dental hygiene: a study of tutor actions. AB - This case study illuminates contextual factors related to the tutor experience when senior students served as tutors for sophomore students in a problem-based learning (PBL) course in a baccalaureate dental hygiene program. Data were collected using various sources and methods. Tutors and administrators were interviewed, those tutored completed an anonymous questionnaire, the tutorial process and tutor training sessions were observed, and related documents were examined. Data analysis included open and axial coding, creation of tutor profiles, and identification of patterns. Tutor training included experiencing the PBL student role, attending class, and weekly seminar sessions facilitated by a tutor supervisor. Analysis revealed that tutor behaviors could be distinguished by the nature of intended actions (e.g., telling, asking, clarifying, acknowledging), emphasis of comments (process, content, social), and facilitation style (directive, suggestive, empowering). Patterns in tutor behavior and attitudes emerged related to comfort or growth and persistence or lenience. Differences in tutor understanding and perception of their role and the purpose of PBL appeared to influence the role the tutor assumed. Other factors that influenced tutor behavior included tutor intentions, tutor training, and environmental factors (e.g., time). The study, incorporating Fishbein's integrative model, suggests points of influence on tutor behaviors. PMID- 21642528 TI - Comparative use of podcasts vs. lecture transcripts as learning aids for dental students. AB - The purpose of this project was to describe dental students' use of lecture podcasts versus written lecture transcripts as learning aids under three different circumstances: studying for an exam, reviewing an attended lecture, and reviewing a missed lecture. Additional analyses were performed to see whether demographic differences (e.g., age, gender, language skills, and computer skills) or grade differences were associated with preferences for using podcast versus written lecture transcripts of class notes. Fifty-one percent (n=171) of the second-year dental students at the New York University College of Dentistry voluntarily participated in this survey. The major findings were that 1) a high percentage of students (70-92 percent) used one or both aids in all three utilization circumstances with a consistent preference for podcast use, especially when reviewing a missed lecture; 2) course grades were not associated with the preferred use of either lecture aid; and 3) over half the students listened to the podcasts at speeds that were one and one-half or two times faster than normal speech, especially younger students. Further studies are warranted to delve into the current student generation's preferred learning styles and the resultant learning outcomes associated with those preferences. PMID- 21642529 TI - Development and evaluation of an interactive dental video game to teach dentin bonding. AB - Written and clinical tests compared the change in clinical knowledge and practical clinical skill of first-year dental students watching a clinical video recording of the three-step etch-and-rinse resin bonding system to those using an interactive dental video game teaching the same procedure. The research design was a randomized controlled trial with eighty first-year dental students enrolled in the preclinical operative dentistry course. Students' change in knowledge was measured through written examination using a pre-test and a post-test, as well as clinical tests in the form of a benchtop shear bond strength test. There was no statistically significant difference between teaching methods in regards to change in either knowledge or clinical skills, with one minor exception relating to the wetness of dentin following etching. Students expressed their preference for an interactive self-paced method of teaching. PMID- 21642530 TI - Improving tobacco dependence education among the Loma Linda University School of Dentistry faculty. AB - Tobacco-related health problems are among the most preventable forms of illness. By assuming proactive tobacco use counseling roles, dental professionals can help reduce the number of people who use tobacco. Minimum standards for intervention by dental care providers were established more than a decade ago by the American Dental Association and the American Dental Hygienists' Association. The goal of Loma Linda University School of Dentistry in its tobacco-cessation efforts is to move beyond those standards towards more effective interventions. The school conducted a study to determine the formal education of the faculty, evaluate the current state of tobacco dependence education (TDE) delivered to students, identify topics that dental faculty members wanted to further their education, promote tobacco dependence education among the dental faculty, and enhance teaching moments on the clinic floor. A fifty-seven question survey was e-mailed to all faculty members with >0.4 FTE (full-time equivalent) during the 2007-08 school year. The response rate was 80 percent (101 out of 126). The results revealed that faculty members have limited formal training; however, 73.1 percent agreed that TDE would be beneficial to them. They also believed that, upon graduation, dental professionals should be able to perform at least a ten-minute moderate intervention program and discuss options for tobacco dependence treatments with patients. This project was designed to establish a 2008-09 baseline of TDE clinical practices, knowledge, and attitudes and to assess the effects of faculty development, curricular didactic, and clinical changes. PMID- 21642531 TI - Multiplexed RNA structure characterization with selective 2'-hydroxyl acylation analyzed by primer extension sequencing (SHAPE-Seq). AB - New regulatory roles continue to emerge for both natural and engineered noncoding RNAs, many of which have specific secondary and tertiary structures essential to their function. Thus there is a growing need to develop technologies that enable rapid characterization of structural features within complex RNA populations. We have developed a high-throughput technique, SHAPE-Seq, that can simultaneously measure quantitative, single nucleotide-resolution secondary and tertiary structural information for hundreds of RNA molecules of arbitrary sequence. SHAPE Seq combines selective 2'-hydroxyl acylation analyzed by primer extension (SHAPE) chemistry with multiplexed paired-end deep sequencing of primer extension products. This generates millions of sequencing reads, which are then analyzed using a fully automated data analysis pipeline, based on a rigorous maximum likelihood model of the SHAPE-Seq experiment. We demonstrate the ability of SHAPE Seq to accurately infer secondary and tertiary structural information, detect subtle conformational changes due to single nucleotide point mutations, and simultaneously measure the structures of a complex pool of different RNA molecules. SHAPE-Seq thus represents a powerful step toward making the study of RNA secondary and tertiary structures high throughput and accessible to a wide array of scientific pursuits, from fundamental biological investigations to engineering RNA for synthetic biological systems. PMID- 21642532 TI - Evolutionarily conserved surface residues constitute actin binding sites of tropomyosin. AB - Tropomyosin (Tm) is a two-chained, alpha-helical coiled-coil protein that associates end-to-end to form a continuous strand along actin filaments and regulates the functions and stability of actin in eukaryotic muscle and nonmuscle cells. Mutations in Tm cause skeletal and cardiac myopathies. We applied a neoteric molecular evolution approach to gain insight into the fundamental unresolved question of what makes the Tm coiled coil an actin binding protein. We carried out a phylogenetic analysis of 70 coding sequences of Tm genes from 26 animal species, from cnidarians to chordates, and evaluated the substitution rates (omega) at individual codons to identify conserved sites. The most conserved residues at surface b, c, f heptad repeat positions were mutated in rat striated muscle alphaTm and expressed in Escherichia coli. Each mutant had 3-4 sites mutated to Ala within the first half or the second half of periods 2-6. Actin affinity and thermodynamic stability were determined in vitro. Mutations in the first half of periods 2, 4, and 5 resulted in the largest reduction in actin affinity (> 4-fold), indicating these mutations include residues in actin-binding sites. Mutations in the second half of the periods had a <= 2-fold effect on affinity indicating these residues may be involved in other conserved regulatory functions. The structural relevance of these results was assessed by constructing molecular models for the actin-Tm filament. Molecular evolution analysis is a general approach that may be used to identify potential binding sites of a protein for a conserved protein. PMID- 21642533 TI - Bidirectional plasticity in the primate inferior olive induced by chronic ethanol intoxication and sustained abstinence. AB - The brain adapts to chronic ethanol intoxication by altering synaptic and ion channel function to increase excitability, a homeostatic counterbalance to inhibition by alcohol. Delirium tremens occurs when those adaptations are unmasked during withdrawal, but little is known about whether the primate brain returns to normal with repeated bouts of ethanol abuse and abstinence. Here, we show a form of bidirectional plasticity of pacemaking currents induced by chronic heavy drinking within the inferior olive of cynomolgus monkeys. Intracellular recordings of inferior olive neurons demonstrated that ethanol inhibited the tail current triggered by release from hyperpolarization (I(tail)). Both the slow deactivation of hyperpolarization-activated cyclic nucleotide-gated channels conducting the hyperpolarization-activated inward current and the activation of Ca(v)3.1 channels conducting the T-type calcium current (I(T)) contributed to I(tail), but ethanol inhibited only the I(T) component of I(tail). Recordings of inferior olive neurons obtained from chronically intoxicated monkeys revealed a significant up-regulation in I(tail) that was induced by 1 y of daily ethanol self-administration. The up-regulation was caused by a specific increase in I(T) which (i) greatly increased neurons' susceptibility for rebound excitation following hyperpolarization and (ii) may have accounted for intention tremors observed during ethanol withdrawal. In another set of monkeys, sustained abstinence produced the opposite effects: (i) a reduction in rebound excitability and (ii) a down-regulation of I(tail) caused by the down-regulation of both the hyperpolarization-activated inward current and I(T). Bidirectional plasticity of two hyperpolarization-sensitive currents following chronic ethanol abuse and abstinence may underlie persistent brain dysfunction in primates and be a target for therapy. PMID- 21642534 TI - Structural basis for the neutralization and genotype specificity of hepatitis E virus. AB - Hepatitis E virus (HEV) causes acute hepatitis in humans, predominantly by contamination of food and water, and is characterized by jaundice and flu-like aches and pains. To date, no vaccines are commercially available to prevent the disease caused by HEV. Previously, we showed that a monoclonal antibody, 8C11, specifically recognizes a neutralizing conformational epitope on HEV genotype I. The antibody 8C11 blocks the virus-like particle from binding to and penetrating the host cell. Here, we report the complex crystal structure of 8C11 Fab with HEV E2s(I) domain at 1.9 A resolution. The 8C11 epitopes on E2s(I) were identified at Asp(496)-Thr(499), Val(510)-Leu(514), and Asn(573)-Arg(578). Mutations and cell model assays identified Arg(512) as the most crucial residue for 8C11 interaction with and neutralization of HEV. Interestingly, 8C11 specifically neutralizes HEV genotype I, but not the other genotypes. Because HEV type I and IV are the most abundant genotypes, to understand this specificity further we determined the structure of E2s(IV) at 1.79 A resolution and an E2s(IV) complex with 8C11 model was generated. The comparison between the 8C11 complexes with type I and IV revealed the key residues that distinguish these two genotypes. Of particular interest, the residue at amino acid position 497 at the 8C11 epitope region of E2s is distinct among these two genotypes. Swapping this residue from one genotype to another inversed the 8C11 reactivity, demonstrating the essential role played by amino acid 497 in the genotype recognition. These studies may lead to the development of antibody-based drugs for the specific treatment against HEV. PMID- 21642535 TI - Homologous recombination research is heating up and ready for therapy. PMID- 21642536 TI - Modeling and automation of sequencing-based characterization of RNA structure. AB - Sequence census methods reduce molecular measurements such as transcript abundance and protein-nucleic acid interactions to counting problems via DNA sequencing. We focus on a novel assay utilizing this approach, called selective 2'-hydroxyl acylation analyzed by primer extension sequencing (SHAPE-Seq), that can be used to characterize RNA secondary and tertiary structure. We describe a fully automated data analysis pipeline for SHAPE-Seq analysis that includes read processing, mapping, and structural inference based on a model of the experiment. Our methods rely on the solution of a series of convex optimization problems for which we develop efficient and effective numerical algorithms. Our results can be easily extended to other chemical probes of RNA structure, and also generalized to modeling polymerase drop-off in other sequence census-based experiments. PMID- 21642537 TI - Structural model of the TRPP2/PKD1 C-terminal coiled-coil complex produced by a combined computational and experimental approach. AB - Autosomal dominant polycystic kidney disease (ADPKD) is caused by mutations in TRPP2 and PKD1, which form an ion channel/receptor complex containing three TRPP2 and one PKD1. A TRPP2 C-terminal coiled-coil trimer, critical for the assembly of this complex, associates with a single PKD1 C-terminal coiled-coil. Many ADPKD pathogenic mutations result in the abolishment of the TRPP2/PKD1 coiled-coil complex. To gain molecular and functional insights into this heterotetrameric complex, we computationally constructed a structural model by using a two-step docking strategy, based on a known crystal structure of the TRPP2 coiled-coil trimer. The model shows that this tetrameric complex has a novel di-trimer configuration: An upstream trimer made of three TRPP2 helices and a downstream trimer made of two TRPP2 helices and one PKD1 helix. Mutagenesis and biochemical analysis identified critical TRPP2/PKD1 interface contacts essential for the heteromeric coiled-coil complex. Mutation of these interface positions in the full-length proteins showed that these interactions were critical for the assembly of the full-length complex in cells. Our results provide a means to specifically weaken the TRPP2 and PKD1 association, thus facilitating future in vitro and in vivo studies on the functional importance of this association. PMID- 21642538 TI - Convexity of quantum chi2-divergence. AB - The general quantum chi(2)-divergence has recently been introduced by Temme et al. [Temme K, Kastoryano M, Ruskai M, Wolf M, Verstrate F (2010) J Math Phys 51:122201] and applied to quantum channels (quantum Markov processes). The quantum chi(2)-divergence is not unique, as opposed to the classical chi(2) divergence, but depends on the choice of quantum statistics. It was noticed that the elements in a particular one-parameter family of quantum chi(2)-divergences are convex functions in the density matrices (rho,sigma), thus mirroring the convexity of the classical chi(2)(p,q)-divergence in probability distributions (p,q). We prove that any quantum chi(2)-divergence is a convex function in its two arguments. PMID- 21642539 TI - Binding of flavivirus nonstructural protein NS1 to C4b binding protein modulates complement activation. AB - The complement system plays a pivotal protective role in the innate immune response to many pathogens including flaviviruses. Flavivirus nonstructural protein 1 (NS1) is a secreted nonstructural glycoprotein that accumulates in plasma to high levels and is displayed on the surface of infected cells but absent from viral particles. Previous work has defined an immune evasion role of flavivirus NS1 in limiting complement activation by forming a complex with C1s and C4 to promote cleavage of C4 to C4b. In this study, we demonstrate a second mechanism, also involving C4 and its active fragment C4b, by which NS1 antagonizes complement activation. Dengue, West Nile, or yellow fever virus NS1 directly associated with C4b binding protein (C4BP), a complement regulatory plasma protein that attenuates the classical and lectin pathways. Soluble NS1 recruited C4BP to inactivate C4b in solution and on the plasma membrane. Mapping studies revealed that the interaction sites of NS1 on C4BP partially overlap with the C4b binding sites. Together, these studies further define the immune evasion potential of NS1 in reducing the functional capacity of C4 in complement activation and control of flavivirus infection. PMID- 21642540 TI - Granule exocytosis contributes to priming and activation of the human neutrophil respiratory burst. AB - The role of exocytosis in the human neutrophil respiratory burst was determined using a fusion protein (TAT-SNAP-23) containing the HIV transactivator of transcription (TAT) cell-penetrating sequence and the N-terminal SNARE domain of synaptosome-associated protein-23 (SNAP-23). This agent inhibited stimulated exocytosis of secretory vesicles and gelatinase and specific granules but not azurophil granules. GST pulldown showed that TAT-SNAP-23 bound to the combination of vesicle-associated membrane protein-2 and syntaxin-4 but not to either individually. TAT-SNAP-23 reduced phagocytosis-stimulated hydrogen peroxide production by 60% without affecting phagocytosis or generation of HOCl within phagosomes. TAT-SNAP-23 had no effect on fMLF-stimulated superoxide release but significantly inhibited priming of this response by TNF-alpha and platelet activating factor. Pretreatment with TAT-SNAP-23 inhibited the increase in plasma membrane expression of gp91(phox) in TNF-alpha-primed neutrophils, whereas TNF alpha activation of ERK1/2 and p38 MAPK was not affected. The data demonstrate that neutrophil granule exocytosis contributes to phagocytosis-induced respiratory burst activity and plays a critical role in priming of the respiratory burst by increasing expression of membrane components of the NADPH oxidase. PMID- 21642541 TI - NK cells promote type 1 T cell immunity through modulating the function of dendritic cells during intracellular bacterial infection. AB - Dendritic cells (DC) play a key role in establishing protective adaptive immunity in intracellular bacterial infections, but the cells influencing DC function in vivo remain unclear. In this study, we investigated the role of NK cells in modulating the function of DC using a murine Chlamydia infection model. We found that the NK cell-depleted mice showed exacerbated disease after respiratory tract Chlamydia muridarum infection, which was correlated with altered T cell cytokine profile. Furthermore, DC from C. muridarum-infected NK-depleted mice (NK(-)DC) exhibited a less mature phenotype compared with that of DC from the infected mice without NK depletion (NK(+)DC). NK(-)DC produced significantly lower levels of both IL-12 and IL-10 than those of NK(+)DC. Moreover, NK(-)DC showed reduced ability to direct primary and established Ag-specific Th1 CD4(+) T cell responses in DC-T coculture systems. More importantly, adoptive transfer of NK(-)DC, in contrast to NK(+)DC, failed to induce type 1 protective immunity in recipients after challenge infection. Finally, NK cells showed strong direct enhancing effect on IL-12 production by DC in an NK-DC coculture system, which was partially reduced by blocking NKG2D receptors signaling and virtually abolished by neutralizing IFN-gamma activity. The data demonstrate a critical role of NK cells in modulating DC function in an intracellular bacterial infection. PMID- 21642542 TI - Knockdown of HMGB1 in tumor cells attenuates their ability to induce regulatory T cells and uncovers naturally acquired CD8 T cell-dependent antitumor immunity. AB - Although high mobility group box 1 (HMGB1) in tumor cells is involved in many aspects of tumor progression, its role in tumor immune suppression remains elusive. Host cell-derived IL-10 suppressed a naturally acquired CD8 T cell dependent antitumor response. The suppressive activity of tumor-associated Foxp3(+)CD4(+)CD25(+) regulatory T cells (Treg) was IL-10 dependent. Neutralizing HMGB1 impaired tumor cell-promoted IL-10 production by Treg. Short hairpin RNA mediated knockdown of HMGB1 (HMGB1 KD) in tumor cells did not affect tumor cell growth but uncovered naturally acquired long-lasting tumor-specific IFN-gamma- or TNF-alpha-producing CD8 T cell responses and attenuated their ability to induce Treg, leading to naturally acquired CD8 T cell- or IFN-gamma-dependent tumor rejection. The data suggest that tumor cell-derived HMGB1 may suppress naturally acquired CD8 T cell-dependent antitumor immunity via enhancing Treg to produce IL 10, which is necessary for Treg-mediated immune suppression. PMID- 21642543 TI - Alternative pathway activation of complement by Shiga toxin promotes exuberant C3a formation that triggers microvascular thrombosis. AB - Shiga toxin (Stx)-producing E.coli O157:H7 has become a global threat to public health; it is a primary cause of diarrhea-associated hemolytic uremic syndrome (HUS), a disorder of thrombocytopenia, microangiopathic hemolytic anemia, and acute renal failure with thrombi occluding renal microcirculation. In this study, we explored whether Stx triggers complement-dependent microvascular thrombosis in in vitro and in vivo experimental settings of HUS. Stx induced on human microvascular endothelial cell surface the expression of P-selectin, which bound and activated C3 via the alternative pathway, leading to thrombus formation under flow. In the search for mechanisms linking complement activation and thrombosis, we found that exuberant complement activation in response to Stx generated an increased amount of C3a that caused further endothelial P-selectin expression, thrombomodulin (TM) loss, and thrombus formation. In a murine model of HUS obtained by coinjection of Stx2 and LPS and characterized by thrombocytopenia and renal dysfunction, upregulation of glomerular endothelial P-selectin was associated with C3 and fibrin(ogen) deposits, platelet clumps, and reduced TM expression. Treatment with anti-P-selectin Ab limited glomerular C3 accumulation. Factor B-deficient mice after Stx2/LPS exhibited less thrombocytopenia and were protected against glomerular abnormalities and renal function impairment, indicating the involvement of complement activation via the alternative pathway in the glomerular thrombotic process in HUS mice. The functional role of C3a was documented by data showing that glomerular fibrin(ogen), platelet clumps, and TM loss were markedly decreased in HUS mice receiving C3aR antagonist. These results identify Stx-induced complement activation, via P-selectin, as a key mechanism of C3a-dependent microvascular thrombosis in diarrhea-associated HUS. PMID- 21642544 TI - Transcriptional control of rapid recall by memory CD4 T cells. AB - Memory T cells are distinguished from naive T cells by their rapid production of effector cytokines, although mechanisms for this recall response remain undefined. In this study, we investigated transcriptional mechanisms for rapid IFN-gamma production by Ag-specific memory CD4 T cells. In naive CD4 T cells, IFN gamma production only occurred after sustained Ag activation and was associated with high expression of the T-bet transcription factor required for Th1 differentiation and with T-bet binding to the IFN-gamma promoter as assessed by chromatin immunoprecipitation analysis. By contrast, immediate IFN-gamma production by Ag-stimulated memory CD4 T cells occurred in the absence of significant nuclear T-bet expression or T-bet engagement on the IFN-gamma promoter. We identified rapid induction of NF-kappaB transcriptional activity and increased engagement of NF-kappaB on the IFN-gamma promoter at rapid times after TCR stimulation of memory compared with naive CD4 T cells. Moreover, pharmacologic inhibition of NF-kappaB activity or peptide-mediated inhibition of NF-kappaB p50 translocation abrogated early memory T cell signaling and TCR mediated effector function. Our results reveal a molecular mechanism for memory T cell recall through enhanced NF-kappaB p50 activation and promoter engagement, with important implications for memory T cell modulation in vaccines, autoimmunity, and transplantation. PMID- 21642545 TI - Connexin 43 signaling enhances the generation of Foxp3+ regulatory T cells. AB - Despite their importance for the functioning of the immune system, thymic development and peripheral maintenance of Foxp3(+) regulatory T (T(R)) cells are poorly understood. We have found that connexin 43 (Cx43), expressed by thymic T(R) cells progenitors, supports T(R) development. Mice with deletion of the Cx43 gene induced in T cells produce only few T(R) cells and had increased proportion of activated T cells in the lymph nodes, suggesting impaired peripheral tolerance. Reduction of the T(R) cell numbers was accompanied by increased presence of CD4(+)CD25(+)GITR(+)Foxp3(-) T cells, which did not produce inflammatory cytokines and lost suppressor function. These results strongly argue that we have discovered a novel signaling pathway, controlled by Cx43, that enhances the generation of T(R) cells. We propose that a possible mechanism of Cx43 activity is by regulating Foxp3 expression in T(R) lineage cells. PMID- 21642546 TI - Targeted mRNA oxidation regulates sunflower seed dormancy alleviation during dry after-ripening. AB - After-ripening is the mechanism by which dormant seeds become nondormant during their dry storage after harvest. The absence of free water in mature seeds does not allow detectable metabolism; thus, the processes associated with dormancy release under these conditions are largely unknown. We show here that sunflower (Helianthus annuus) seed alleviation of dormancy during after-ripening is associated with mRNA oxidation and that this oxidation is prevented when seeds are maintained dormant. In vitro approaches demonstrate that mRNA oxidation results in artifacts in cDNA-amplified fragment length polymorphim analysis and alters protein translation. The oxidation of transcripts is not random but selective, and, using microarrays, we identified 24 stored mRNAs that became highly oxidized during after-ripening. Oxidized transcripts mainly correspond to genes involved in responses to stress and in cell signaling. Among them, protein phosphatase 2C PPH1, mitogen-activated protein kinase phosphatase 1, and phenyl ammonia lyase 1 were identified. We propose that targeted mRNA oxidation during dry after-ripening of dormant seeds could be a process that governs cell signaling toward germination in the early steps of seed imbibition. PMID- 21642547 TI - Gibberellin regulates PIN-FORMED abundance and is required for auxin transport dependent growth and development in Arabidopsis thaliana. AB - Plants integrate different regulatory signals to control their growth and development. Although a number of physiological observations suggest that there is crosstalk between the phytohormone gibberellin (GA) and auxin, as well as with auxin transport, the molecular basis for this hormonal crosstalk remains largely unexplained. Here, we show that auxin transport is reduced in the inflorescences of Arabidopsis thaliana mutants deficient in GA biosynthesis and signaling. We further show that this reduced auxin transport correlates with a reduction in the abundance of PIN-FORMED (PIN) auxin efflux facilitators in GA-deficient plants and that PIN protein levels recover to wild-type levels following GA treatment. We also demonstrate that the regulation of PIN protein levels cannot be explained by a transcriptional regulation of the PIN genes but that GA deficiency promotes, at least in the case of PIN2, the targeting of PIN proteins for vacuolar degradation. In genetic studies, we reveal that the reduced auxin transport of GA mutants correlates with an impairment in two PIN-dependent growth processes, namely, cotyledon differentiation and root gravitropic responses. Our study thus presents evidence for a role of GA in these growth responses and for a GA dependent modulation of PIN turnover that may be causative for these differential growth responses. PMID- 21642549 TI - Malonyl-CoA synthetase, encoded by ACYL ACTIVATING ENZYME13, is essential for growth and development of Arabidopsis. AB - Malonyl-CoA is the precursor for fatty acid synthesis and elongation. It is also one of the building blocks for the biosynthesis of some phytoalexins, flavonoids, and many malonylated compounds. In plants as well as in animals, malonyl-CoA is almost exclusively derived from acetyl-CoA by acetyl-CoA carboxylase (EC 6.4.1.2). However, previous studies have suggested that malonyl-CoA may also be made directly from malonic acid by malonyl-CoA synthetase (EC 6.2.1.14). Here, we report the cloning of a eukaryotic malonyl-CoA synthetase gene, Acyl Activating Enzyme13 (AAE13; At3g16170), from Arabidopsis thaliana. Recombinant AAE13 protein showed high activity against malonic acid (K(m) = 529.4 +/- 98.5 MUM; V(m) = 24.0 +/- 2.7 MUmol/mg/min) but little or no activity against other dicarboxylic or fatty acids tested. Exogenous malonic acid was toxic to Arabidopsis seedlings and caused accumulation of malonic and succinic acids in the seedlings. aae13 null mutants also grew poorly and accumulated malonic and succinic acids. These defects were complemented by an AAE13 transgene or by a bacterial malonyl-CoA synthetase gene under control of the AAE13 promoter. Our results demonstrate that the malonyl-CoA synthetase encoded by AAE13 is essential for healthy growth and development, probably because it is required for the detoxification of malonate. PMID- 21642550 TI - The plasma membrane of the cyanobacterium Gloeobacter violaceus contains segregated bioenergetic domains. AB - The light reactions of oxygenic photosynthesis almost invariably take place in the thylakoid membranes, a highly specialized internal membrane system located in the stroma of chloroplasts and the cytoplasm of cyanobacteria. The only known exception is the primordial cyanobacterium Gloeobacter violaceus, which evolved before the appearance of thylakoids and harbors the photosynthetic complexes in the plasma membrane. Thus, studies on G. violaceus not only shed light on the evolutionary origin and the functional advantages of thylakoid membranes but also might include insights regarding thylakoid formation during chloroplast differentiation. Based on biochemical isolation and direct in vivo characterization, we report here structural and functional domains in the cytoplasmic membrane of a cyanobacterium. Although G. violaceus has no internal membranes, it does have localized domains with apparently specialized functions in its plasma membrane, in which both the photosynthetic and the respiratory complexes are concentrated. These bioenergetic domains can be visualized by confocal microscopy, and they can be isolated by a simple procedure. Proteomic analysis of these domains indicates their physiological function and suggests a protein sorting mechanism via interaction with membrane-intrinsic terpenoids. Based on these results, we propose specialized domains in the plasma membrane as evolutionary precursors of thylakoids. PMID- 21642551 TI - How do speakers resist distraction? Evidence from a taboo picture-word interference task. AB - Even in the presence of irrelevant stimuli, word production is a highly accurate and fluent process. But how do speakers prevent themselves from naming the wrong things? One possibility is that an attentional system inhibits task-irrelevant representations. Alternatively, a verbal self-monitoring system might check speech for accuracy and remove errors stemming from irrelevant information. Because self-monitoring is sensitive to social appropriateness, taboo errors should be intercepted more than neutral errors are. To prevent embarrassment, speakers might also speak more slowly when confronted with taboo distractors. Our results from two experiments are consistent with the self-monitoring account: Examining picture-naming speed (Experiment 1) and accuracy (Experiment 2), we found fewer naming errors but longer picture-naming latencies for pictures presented with taboo distractors than for pictures presented with neutral distractors. These results suggest that when intrusions of irrelevant words are highly undesirable, speakers do not simply inhibit these words: Rather, the language-production system adjusts itself to the context and filters out the undesirable words. PMID- 21642548 TI - Coping with stresses: roles of calcium- and calcium/calmodulin-regulated gene expression. AB - Abiotic and biotic stresses are major limiting factors of crop yields and cause billions of dollars of losses annually around the world. It is hoped that understanding at the molecular level how plants respond to adverse conditions and adapt to a changing environment will help in developing plants that can better cope with stresses. Acquisition of stress tolerance requires orchestration of a multitude of biochemical and physiological changes, and most of these depend on changes in gene expression. Research during the last two decades has established that different stresses cause signal-specific changes in cellular Ca(2+) level, which functions as a messenger in modulating diverse physiological processes that are important for stress adaptation. In recent years, many Ca(2+) and Ca(2+)/calmodulin (CaM) binding transcription factors (TFs) have been identified in plants. Functional analyses of some of these TFs indicate that they play key roles in stress signaling pathways. Here, we review recent progress in this area with emphasis on the roles of Ca(2+)- and Ca(2+)/CaM-regulated transcription in stress responses. We will discuss emerging paradigms in the field, highlight the areas that need further investigation, and present some promising novel high throughput tools to address Ca(2+)-regulated transcriptional networks. PMID- 21642552 TI - Testing the role of adolescent sexual initiation in later-life sexual risk behavior: a longitudinal twin design. AB - The consistent association between adolescent sexual initiation (ASI) and risky adult sexual behavior (RASB) has generally been assumed to indicate that ASI has a causal effect on RASB; consequently, it is assumed that delaying ASI will reduce RASB. Yet the ASI-RASB association might be better accounted for by some third variable. We evaluated the causal role of ASI (initiation of oral, anal, or vaginal sex at or before age 16) in influencing RASB in a longitudinal sample of 2,173 twins (followed from ages 11 to 24 or from ages 17 to 29) using two methods: the discordant-twin design and the propensity-score design. The former controlled for unmeasured genetic and shared environmental factors, and the latter controlled for measured nonshared environmental factors. We replicated the link between ASI and RASB reported in previous research, but results from the discordant-twin and propensity-score analyses suggested that this association is better explained by common genetic or environmental risk factors than as a causal effect. These findings suggest that preventing ASI is unlikely to reduce RASB. PMID- 21642553 TI - Do 18-month-olds really attribute mental states to others? A critical test. AB - In the research reported here, we investigated whether 18-month-olds would use their own past experience of visual access to attribute perception and consequent beliefs to other people. Infants in this study wore either opaque blindfolds (opaque condition) or trick blindfolds that looked opaque but were actually transparent (trick condition). Then both groups of infants observed an actor wearing one of the same blindfolds that they themselves had experienced, while a puppet removed an object from its location. Anticipatory eye movements revealed that infants who had experienced opaque blindfolds expected the actor to behave in accordance with a false belief about the object's location, but that infants who had experienced trick blindfolds did not exhibit that expectation. Our results suggest that 18-month-olds used self-experience with the blindfolds to assess the actor's visual access and to update her belief state accordingly. These data constitute compelling evidence that 18-month-olds infer perceptual access and appreciate its causal role in altering the epistemic states of other people. PMID- 21642554 TI - Advancing pharmacy practice models: achieving consensus. PMID- 21642557 TI - Home-based medication management system looks to future. PMID- 21642558 TI - Linagliptin approved for type 2 diabetes. PMID- 21642559 TI - CMS reveals Medicare's value-based purchasing program for hospitals. PMID- 21642560 TI - Fairleigh Dickinson, Medco combine forces to modernize pharmacy practice. PMID- 21642561 TI - New practitioners and the Pharmacy Practice Model Initiative: our opportunity to define the future. PMID- 21642562 TI - Three signals from the Pharmacy Practice Model Summit. PMID- 21642564 TI - Needs and opportunities for achieving optimal outcomes from the use of medicines in hospitals and health systems. PMID- 21642565 TI - Health care imperative for practice model change. PMID- 21642566 TI - Changing the pharmacy practice model: a health-system executive's view. PMID- 21642567 TI - Health-system pharmacy's imperative for practice model change. PMID- 21642568 TI - Implementing practice model change: Opportunities and challenges in a changing environment. PMID- 21642569 TI - Critical requirements for health-system pharmacy practice models that achieve optimal use of medicines. PMID- 21642570 TI - Cutting-edge practice model: experience in a Veterans Affairs hospital. PMID- 21642571 TI - Cutting-edge practice model: an integrated model within a large academic medical center. PMID- 21642572 TI - Cutting-edge practice model: experience in a small community hospital. PMID- 21642573 TI - Cutting-edge practice model: a mixed integrated and clinical specialist model in a large health system. PMID- 21642574 TI - Cutting-edge practice model: Experience in a community hospital system. PMID- 21642575 TI - Opportunities and challenges related to technology in supporting optimal pharmacy practice models in hospitals and health systems. PMID- 21642576 TI - Safe, effective, and efficient medication distribution: Challenging paradigms and bedrocks. PMID- 21642577 TI - Opportunities and challenges related to pharmacy technicians in supporting optimal pharmacy practice models in health systems. PMID- 21642578 TI - Methods of fostering change in the practice model at the pharmacy department level. PMID- 21642579 TI - Strategies for success in implementing practice model change. PMID- 21642580 TI - The consensus of the Pharmacy Practice Model Summit. PMID- 21642581 TI - Reflections on the key messages of the Pharmacy Practice Model Summit. PMID- 21642582 TI - ASHP's commitment to keeping the Pharmacy Practice Model Summit alive. PMID- 21642585 TI - Agonist-promoted homologous desensitization of human airway smooth muscle bitter taste receptors. AB - Bitter taste receptors (TAS2Rs) were shown to be expressed in human airway smooth muscle (ASM). They couple to specialized [Ca(2+)](i) release, leading to membrane hyperpolarization, the relaxation of ASM, and marked bronchodilation. TAS2Rs are G-protein-coupled receptors, known to undergo rapid agonist-promoted desensitization that can limit therapeutic efficacy. Because TAS2Rs represent a new drug target for treating obstructive lung disease, we investigated their capacity for rapid desensitization, and assessed their potential mechanisms. The pretreatment of human ASM cells with the prototypic TAS2R agonist quinine resulted in a 31% +/- 5.1% desensitization of the [Ca(2+)](i) response from a subsequent exposure to quinine. No significant change in the endothelin stimulated [Ca(2+)](i) response was attributed to the short-term use of quinine, indicating a homologous form of desensitization. The TAS2R agonist saccharin also evoked desensitization, and cross-compound desensitization with quinine was evident. Desensitization of the [Ca(2+)](i) response was attenuated by a dynamin inhibitor, suggesting that receptor internalization (a G-protein coupled receptor kinase [GRK]-mediated, beta-arrestin-mediated process) plays an integral role in the desensitization of TAS2R. Desensitization was insensitive to antagonists of the second messenger kinases protein kinase A and protein kinase C. Using intact airways, short-term, agonist-promoted TAS2R desensitization of the relaxation response was also observed. Thus these receptors, which represent a potential novel target for direct bronchodilators, undergo a modest degree of agonist promoted desensitization that may affect clinical efficacy. Collectively, the results of these mechanistic studies, along with the multiple serines and threonines in intracellular loop 3 and the cytoplasmic tail of TAS2Rs, suggest a GRK-mediated mode of desensitization. PMID- 21642586 TI - Fungal allergen beta-glucans trigger p38 mitogen-activated protein kinase mediated IL-6 translation in lung epithelial cells. AB - In addition to immune cells, airway epithelial cells can contribute to and shape the immune response in the lung by secreting specific cytokines. IL-6 is a key factor in determining the effector fate of CD4(+) T cells. Here we show that under basal conditions, the IL-6 gene is already highly expressed in lung epithelial cells, but not in immune cells resident in the lung. However, upon exposure of the lungs to fungal allergens, the direct contact of beta-glucans present in the fungus cell wall with lung epithelial cells is sufficient to trigger the rapid synthesis and secretion of IL-6 protein. This posttranscriptional regulation of IL-6 in response to fungal extracts is mediated by the p38 mitogen-activated protein kinase pathway. The inhalation of beta glucans with a nonallergenic antigen is sufficient to provide an adjuvant effect that leads to mucous hyperplasia in the airways. Thus, beta-glucans may constitute a common determinant of the fungal and plant-derived allergens responsible for some of the pathological features in allergic asthma. PMID- 21642587 TI - Intrinsic phenotypic differences of asthmatic epithelium and its inflammatory responses to respiratory syncytial virus and air pollution. AB - A substantial proportion of healthcare cost associated with asthma is attributable to exacerbations of the disease. Within the airway, the epithelium forms the mucosal immune barrier, the first structural cell defense against common environmental insults such as respiratory syncytial virus (RSV) and particulate matter. We sought to characterize the phenotype of differentiated asthmatic-derived airway epithelial cultures and their intrinsic inflammatory responses to environmental challenges. Air-liquid interface (ALI) cultures were generated from asthmatic (n = 6) and nonasthmatic (n = 6) airway epithelial cells. Airway tissue and ALI cultures were analyzed by immunohistochemistry for cytokeratin-5, E-cadherin, Ki67, Muc5AC, NF-kappaB, the activation of p38, and apoptosis. ALI cultures were exposed to RSV (4 * 10(6) plaque forming unit/ml), particulate matter collected by Environmental Health Canada (EHC-93, 100 MUg/ml), or mechanically wounded for 24, 48, and 96 hours and basolateral supernatants analyzed for inflammatory cytokines, using Luminex and ELISA. The airway epithelium in airway sections of patients with asthma as well as in vitro ALI cultures demonstrated a less differentiated epithelium, characterized by elevated numbers of basal cells marked by the expression of cytokeratin-5, increased phosphorylation of p38 mitogen-activated protein kinase, and less adherens junction protein E-cadherin. Transepithelial resistance was not different between asthmatic and nonasthmatic cultures. In response to infection with RSV, exposure to EHC-93, or mechanical wounding, asthmatic ALI cultures released greater concentrations of IL-6, IL-8, and granulocyte macrophage colony-stimulating factor, compared with nonasthmatic cultures (P < 0.05). This parallel ex vivo and in vitro study of the asthmatic epithelium demonstrates an intrinsically altered phenotype and aberrant inflammatory response to common environmental challenges, compared with nonasthmatic epithelium. PMID- 21642588 TI - Pneumocystis S-adenosylmethionine transport: a potential drug target. AB - Pneumocystis pneumonia (PCP) is a life-threatening condition in immunosuppressed patients. Current treatments are inadequate, and new drug leads are needed. This fungus depends on its host for S-adenosylmethionine (AdoMet), a critical metabolic intermediate ordinarily synthesized by individual cells as needed. Pneumocystis contains a gene coding for the AdoMet-synthesizing enzyme methionine ATP transferase (MAT), and the protein is expressed. However, the fungus lacks MAT activity, and infection causes the depletion of host plasma AdoMet. The uptake of Pneumocystis AdoMet was shown to be exquisitely specific, which suggests the transport of AdoMet as a potential drug target. Here we report on the discovery of PcPET8, a Pneumocystis gene with homology to mitochondrial AdoMet transporters. When expressed by Saccharomyces cerevisiae, it locates properly to the mitochondrion and complements a strain of S. cerevisiae lacking its native mitochondrial AdoMet transporter. The importance of AdoMet transport is demonstrated by the ability of the AdoMet analogue sinefungin to block the uptake of Pneumocystis AdoMet and inhibit growth in culture. Because PcPET8 is likely critical for Pneumocystis, the yeast construct has potential as a surrogate for testing compounds against Pneumocystis. PMID- 21642589 TI - Infection with influenza virus induces IL-33 in murine lungs. AB - IL-33, a novel IL-1 family member, is crucially expressed and involved in pulmonary diseases, but its regulation in viral diseases such as influenza A virus (IAV) remains unclear. This study aimed to characterize the expression and release of IL-33 in lungs of IAV-infected mice in vivo and in murine respiratory epithelial cells (MLE-15) in vitro. Our results provide evidence of up-regulation of IL-33 mRNA in IAV-infected murine lungs, compared with noninfected control mice. The overexpression of IL-33 was positively correlated with a significant increase in mRNA encoding the proinflammatory cytokines TNF-alpha, IFN-gamma, IL 1beta, and IL-6, and was also associated with an increase in IFN-beta mRNA. A profound overexpression of IL-33 protein was evident in IAV-infected murine lungs and bronchoalveolar lavages of influenza-infected mice, compared with low concentrations in naive lungs in vivo. Immunolocalization highlighted the cellular expression of IL-33 in alveolar epithelial and endothelial cells, along with increased infiltrate cells in virus-infected lungs. Further in vitro experiments showed an induction of IL-33 transcript-in MLE-15 cells and human epithelial cells (A549) infected with different strains of IAV in comparison with noninfected cells. In conclusion, our findings evidenced a profound expression of IL-33 in lungs during both in vivo and in vitro IAV infections, suggesting a role for IL-33 in virus-induced lung infections. PMID- 21642590 TI - Clarithromycin inhibits interleukin-13-induced goblet cell hyperplasia in human airway cells. AB - IL-13 is a T-helper class 2 cytokine that induces goblet cell hyperplasia and mucus production in airway epithelial cells. Because macrolide antibiotics are known to have immunomodulatory and mucoregulatory properties, the aim of this study was to examine the effect of clarithromycin on IL-13-induced goblet cell hyperplasia and mucin hypersecretion in normal human bronchial epithelial (NHBE) cells. NHBE cells were cultured to differentiation at an air-liquid interface with IL-13 plus clarithromycin or vehicle. Histochemical analysis was performed using H&E staining, periodic acid-Schiff (PAS) staining, and MUC5AC immunostaining. MUC5AC synthesis was assayed using RT-PCR and ELISA. Western blotting was used to evaluate signaling pathways. IL-13 significantly increased the number of PAS-positive, MUC5AC-positive goblet cells, and this was significantly attenuated by clarithromycin at concentrations greater than 8 MUg/ml (P < 0.01). Clarithromycin also dose-dependently decreased MUC5AC mRNA expression induced by IL-13 (P < 0.001), and, at 24 MUg/ml, clarithromycin significantly attenuated the amount of MUC5AC protein in cell supernatants (P < 0.01). Western blotting showed that clarithromycin affected IL-13 receptor janus kinase signal transducers, activators of transcription6 (STAT6), and epidermal growth factor receptor mitogen-activated protein kinase signaling and that inhibition of these pathways by clarithromycin decreased goblet cell hyperplasia via nuclear factor-kappaB inactivation. We conclude that clarithromycin inhibits goblet cell hyperplasia and may directly regulate mucus secretion by IL-13 in NHBE cells. PMID- 21642591 TI - Expression and function of a novel variant of estrogen receptor-alpha36 in murine airways. AB - Evidence suggests that estrogen signaling is involved in sex differences in the prevalence rates and control of asthma, but the expression patterns of estrogen receptor variants and estrogen function in the lung are not well established. We investigated the expression of major estrogen receptor variants occurring naturally and after the development of allergen-induced airway hyperreactivity in a murine model of allergic asthma, along with the role of estrogen signaling in small-airway ciliary motion and smooth muscle contraction. Female BALB/c mice were sensitized with ovalbumin, and estrogen receptor expression patterns were examined by immunofluorescence and Western blot analysis. Time-lapse video and photodiode-based displacement measurement systems were used to assess the effects of estrogen signaling on airway ciliary beat frequency and smooth muscle contraction. We found that a novel variant of estrogen receptor (ER)-alpha, ER alpha36, is expressed in airway epithelial and smooth muscle cells. ER-alpha36 was predominately localized on the plasma membranes of airway cells. After sensitization to allergen, the expression levels of ER-alpha36 increased significantly (P < 0.01), whereas the expression of ER-beta and ER-alpha66 did not significantly change. Estrogen treatment in vitro resulted in a rapid increase in airway cilia motion in a dose-dependent fashion, but did not exert any effect on airway smooth muscle contraction. We speculate that the up regulation of estrogen receptor expression associated with allergen-induced airway hyperresponsiveness may constitute a protective mechanism to facilitate the clearance of mucus. The identification and localization of specific estrogen receptor subtypes in the lung could lead to newer therapeutic avenues aimed at addressing sex differences of asthma susceptibility. PMID- 21642592 TI - Role of reactive oxygen and nitrogen species in olfactory epithelial injury by the sulfur mustard analogue 2-chloroethyl ethyl sulfide. AB - The inhalation of sulfur mustard (SM) causes substantial deposition in the nasal region. However, specific injury has not been characterized. 2-chloroethyl ethyl sulfide (CEES) is an SM analogue used to model injury and screen potential therapeutics. After the inhalation of CEES, damage to the olfactory epithelium (OE) was extensive. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling-positive cells were present by 4 hours, and maximal at 18-72 hours. Cleaved caspase 3 immunohistochemistry (IHC) was maximal at 18 hours after the inhalation of 5% CEES. Olfactory marker protein (OMP)-positive olfactory neurons were markedly decreased at 18 hours. IHC-positive cells for 3-nitrotyrosine (3 NT) within epithelium were elevated by 8 hours, waning by 18 hours, and absent by 72 hours. AEOL 10150, a catalytic manganoporphyrin antioxidant, administered both subcutaneously (5 mg/kg) and intranasally (50 MUM, "combined treatment"), decreased OE injury. CEES-induced increases in markers of cell death were decreased by combined treatment involving AEOL 10150. CEES-induced changes in OMP and 3-NT immunostaining were markedly improved by combined treatment involving AEOL 10150. The selective inducible nitric oxide synthase inhibitor 1400W (5 mg/kg, subcutaneous), administered 1 hour after inhalation and thereafter every 4 hours (five doses), also reduced OE damage with improved OMP and 3-NT staining. Taken together, these data indicate that reactive oxygen and nitrogen species are important mediators in CEES-induced nasal injury. PMID- 21642593 TI - Effects of adult day care on daily stress of caregivers: a within-person approach. AB - OBJECTIVES. This article examined exposure to and appraisal of care-related stressors associated with use of adult day services (ADS) by family caregivers of individuals with dementia. METHODS: Using a within-person withdrawal design (A-B A-B), we compared caregivers' exposure to and appraisal of behavior problems on days their relative attended and did not attend ADS. Participants were 121 family caregivers enrolling a relative with dementia in an ADS program. Daily assessments were obtained prior to the person's attending ADS for the first time and after 1 and 2 months of attendance on days the person attended and did not attend ADS. RESULTS: Total exposure to stressors and stress appraisals decreased significantly over time on ADS days compared with non-ADS days. Most of this difference was accounted by the time the person with dementia was away from the caregiver, but there were also significant reductions in behavioral problems during the evening and improved sleep immediately following ADS use. DISCUSSION. ADS use lowered caregivers' exposure to stressors and may improve behavior and sleep for people with dementia on days they have ADS. The study highlights how a within-person design can identify the effects of an intermittent intervention, such as ADS. PMID- 21642594 TI - Generation of a multipathogen-specific T-cell product for adoptive immunotherapy based on activation-dependent expression of CD154. AB - Viral and fungal infections remain a leading cause of mortality in patients after hematopoietic stem cell transplantation (HSCT). Adoptive transfer of multipathogen-specific T cells is promising in restoring immunity and thereby preventing and treating infections, but approaches are currently limited because of time-consuming and laborious procedures. Therefore, we investigated a new strategy to simultaneously select T cells specific for viral and fungal pathogens based on activation-dependent expression of CD154. Single- and multipathogen specific T-cell lines with high specificity for adenovirus (AdV), Epstein-Barr virus (EBV), cytomegalovirus (CMV), Candida albicans, and/or Aspergillus fumigatus could be readily generated within 14 days irrespective of the precursor frequency. The T-cell lines responded reproducibly to endogenously processed antigen and specifically proliferated upon antigenic stimulation. Although isolation based on CD154 favors enrichment of CD4(+) T cells, AdV-, EBV- and CMV specific CD8(+) T cells could be expanded and demonstrated lysis of target cells. Conversely, T cell-mediated alloreactivity was almost abrogated compared with the starting fraction. This selection and/or expansion strategy may form the basis for future adoptive immunotherapy trials in patients at risk for multiple infections and may be translated to other antigens. PMID- 21642595 TI - BAD-LAMP is a novel biomarker of nonactivated human plasmacytoid dendritic cells. AB - The brain and dendritic cell (BAD)-associated lysosome-associated membrane protein (LAMP)-like molecule (BAD-LAMP, c20orf103, UNC-46) is a newly identified member of the family of LAMPs. BAD-LAMP expression in the mouse is confined to neurons. We demonstrate here that in humans, BAD-LAMP can specifically be found in the type I IFN-producing plasmacytoid dendritic cells (pDCs). Human BAD-LAMP is localized in the endoplasmic reticulum-Golgi intermediate compartment (ERGIC) of freshly isolated CD123(+) pDCs and is rapidly lost upon activation by unmethylated cytosine-phosphate-guanine (CpG) oligonucleotides. The restricted pattern of BAD-LAMP expression allows for the rapid identification of normal and leukemic human pDCs in tissues and blood. PMID- 21642596 TI - NFATc3 regulates the transcription of genes involved in T-cell activation and angiogenesis. AB - The nuclear factor of activated T cells (NFAT) family of transcription factors plays important roles in many biologic processes, including the development and function of the immune and vascular systems. Cells usually express more than one NFAT member, raising the question of whether NFATs play overlapping roles or if each member has selective functions. Using mRNA knock-down, we show that NFATc3 is specifically required for IL2 and cyclooxygenase-2 (COX2) gene expression in transformed and primary T cells and for T-cell proliferation. We also show that NFATc3 regulates COX2 in endothelial cells, where it is required for COX2, dependent migration and angiogenesis in vivo. These results indicate that individual NFAT members mediate specific functions through the differential regulation of the transcription of target genes. These effects, observed on short term suppression by mRNA knock-down, are likely to have been masked by compensatory effects in gene-knockout studies. PMID- 21642597 TI - Restriction of HIV-1 replication in macrophages and CD4+ T cells from HIV controllers. AB - How HIV controllers (HICs) maintain undetectable viremia without therapy is unknown. The strong CD8(+) T-cell HIV suppressive capacity found in many, but not all, HICs may contribute to long-lasting viral control. However, other earlier defense mechanisms may be involved. Here, we examined intrinsic HIC cell resistance to HIV-1 infection. After in vitro challenge, monocyte-derived macrophages and anti-CD3-activated CD4(+) T cells from HICs showed low HIV-1 susceptibility. CD4 T-cell resistance was independent of HIV-1 coreceptors and affected also SIVmac infection. CD4(+) T cells from HICs expressed ex vivo higher levels of p21(Waf1/Cip1), which has been involved in the control of HIV-1 replication, than cells from control subjects. However, HIV restriction in anti CD3-activated CD4(+) T cells and macrophages was not associated with p21 expression. Restriction inhibited accumulation of reverse transcripts, leading to reduction of HIV-1 integrated proviruses. The block could be overcome by high viral inocula, suggesting the action of a saturable mechanism. Importantly, cell associated HIV-1 DNA load was extremely low in HICs and correlated with CD4(+) T cell permissiveness to infection. These results point to a contribution of intrinsic cell resistance to the control of infection and the containment of viral reservoir in HICs. PMID- 21642598 TI - Production of the plasma-cell survival factor a proliferation-inducing ligand (APRIL) peaks in myeloid precursor cells from human bone marrow. AB - The bone marrow (BM) is an organ extremely efficient in mediating long-term survival of plasma cells (PCs), ensuring an immune humoral memory. This implies that the BM must provide continuously key PC survival factors. Our results show that the BM is an organ constitutively rich in a proliferation-inducing ligand (APRIL), a member of the tumor necrosis factor superfamily implicated in PC survival. APRIL production is induced during hematopoiesis in myeloid cells by non-lineage-committing factors such as stem cell factor, thrombopoietin, IL-3, and FMS-like tyrosine kinase 3 ligand. Notably, APRIL production, both in the human and mouse systems, peaks in myeloid precursor cells, before dropping in fully mature granulocytes. Myeloid cells secrete APRIL that circulates freely in BM plasma to act on PCs, usually at distance from APRIL production sites. Selective APRIL in vivo antagonism and in vitro coculture experiments further demonstrated that myeloid precursor cells mediates PC survival in an APRIL dependent manner Thus, APRIL production by myeloid precursor cells shows that the 2 main BM functions, hematopoiesis and long-term PC survival, are linked. Such constitutive and high APRIL production may explain why BM mediates long-term PC survival. PMID- 21642599 TI - Epidemiology of patellar tendinopathy in elite male soccer players. AB - BACKGROUND: Patellar tendinopathy is common among athletes in jumping sports and in sports with prolonged repetitive stress of the knee extensor apparatus. The epidemiology in soccer is not well described. PURPOSE: This study was undertaken to investigate and describe the epidemiology of patellar tendinopathy in elite male soccer players and evaluate potential risk factors. STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: Between 2001 and 2009, the authors followed 51 European elite soccer clubs (2229 players) from 3 different cohorts: the Swedish First League cohort (SWE) and Union of European Football Associations (UEFA) Champions League cohort (UCL), both playing on natural grass; and the Artifical Turf cohort (ART), playing on third-generation artificial turf. Individual player exposure in training and matches and time-loss injuries were recorded. RESULTS: In total, 137 patellar tendinopathies were recorded, comprising 1.5% of all injuries and corresponding to an incidence of 0.12 injuries/1000 hours. Each season, 2.4% of players were affected, with most injuries (61%) resulting in absence up to 1 week or less. Twenty percent of tendinopathies were recurrent complaints. No significant difference in season prevalence (odds ratio [OR], 0.93; 95% confidence interval [CI], 0.60-1.44; P = .74) or incidence (rate ratio [RR] 1.20; 95% CI, 0.82-1.75; P = .36) was observed between teams playing on artificial turf and natural grass, respectively. Multivariate logistic regression showed that a high total exposure hours (OR, 1.02 per 10-hour increase; 95% CI, 1.00-1.04; P = .033) was a significant risk factor for patellar tendinopathy, and increased body mass was borderline significant (OR, 1.15 per 5-kg increase; 95% CI, 1.00-1.33; P = .055). In addition, 2 acute partial tendon ruptures were recorded, but no total rupture. CONCLUSION: Although mainly mild in nature, patellar tendinopathy is a fairly common condition in elite soccer and the recurrence rate is high. Exposure to artificial turf did not increase the prevalence or incidence of injury. High total amount of exposure was identified as a risk factor for patellar tendinopathy. PMID- 21642600 TI - Position of the bony bridge of lateral meniscal transplants can affect meniscal extrusion. AB - BACKGROUND: There are several reported causes of midbody extrusion after lateral meniscal allograft transplantation. However, there are no reports studying the correlation between the position of the bony bridge and extrusion of the midbody after meniscal allograft transplantation. HYPOTHESIS: The position of the bony bridge of lateral meniscal allografts can affect meniscal extrusion. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Twenty-three consecutive patients underwent a lateral meniscal allograft transplantation using a fresh frozen graft. The lateral meniscal allograft was prepared with a bony bridge. Postoperative evaluations of the meniscal allografts were performed using follow up magnetic resonance imaging 6 months postoperatively. On the coronal view, extrusion was measured as the distance between the outer edge of the articular cartilage of the lateral tibial plateau and the outer edge of the meniscal allograft. On the axial view, the length of the entire tibial plateau (PL) and distance between the lateral edge of the lateral tibial plateau and center of the bony bridge (CB) were measured. Measurements of CB were divided by measurements of PL. The correlation test between CB/PL and extrusion was performed to determine whether the position of the bony bridge can affect extrusion of the midbody of meniscal allograft. RESULTS: The mean center of the bony bridge of the meniscal allograft (CB/PL) was positioned at 42.3% (range, 36.1%-49%; standard deviation [SD] = 3.6%) from the outer edge of the lateral tibial plateau. The mean extrusion of meniscal allografts was 3.2 mm (range, 0-6.5 mm; SD = 2.3). The amount of extrusion was correlated with the position of the bony bridge of the graft and the Pearson correlation coefficient was -.567 (P = .003). The cut-off percentage above which extrusion did not occur was 42.05%. CONCLUSION: The more closely the center of the bony bridge approached 50% of the entire tibial plateau, the less extrusion of the midbody occurred. Anatomic placement of the bony bridge of lateral meniscal allograft is imperative to prevent extrusion after lateral meniscal allograft transplantation. PMID- 21642601 TI - Hospital variation in missed nursing care. AB - Quality of nursing care across hospitals is variable, and this variation can result in poor patient outcomes. One aspect of quality nursing care is the amount of necessary care that is omitted. This article reports on the extent and type of nursing care missed and the reasons for missed care. The MISSCARE Survey was administered to nursing staff (n = 4086) who provide direct patient care in 10 acute care hospitals. Missed nursing care patterns as well as reasons for missing care (labor resources, material resources, and communication) were common across all hospitals. Job title (ie, registered nurse vs nursing assistant), shift worked, absenteeism, perceived staffing adequacy, and patient work loads were significantly associated with missed care. The data from this study can inform quality improvement efforts to reduce missed nursing care and promote favorable patient outcomes. PMID- 21642602 TI - Cardiac index in term pregnant women in the sitting, lateral, and supine positions: an observational, crossover study. AB - BACKGROUND: Aortocaval compression may affect maternal hemodynamic indices and fetal well-being in various maternal positions. There has been much debate regarding the optimal position for performing neuraxial blockade for labor analgesia and cesarean delivery. We hypothesized that in pregnant women at term, cardiac index (CI) may be improved in the lateral positions as compared with the flexed sitting position. Our primary outcome was to measure CI as assessed by suprasternal Doppler. METHODS: A prospective, observational, crossover study was conducted in 25 ASA physical status I/II women with uncomplicated pregnancies presenting for elective cesarean delivery at term. Hemodynamic indices were measured in 4 positions in random order: supine with a 15-degree left tilt, sitting with neck and hips flexed, and flexed left lateral and flexed right lateral positions. Maternal CIs were measured using a noninvasive suprasternal Doppler device and upper arm noninvasive arterial blood pressure. Umbilical Dopplers were performed simultaneously to measure the fetal heart rate and umbilical artery pulsatility and resistivity indices. RESULTS: CI differed by position (P = 0.01); it was higher in the right lateral position compared with the sitting and supine positions (by 8.8% and 8.1%, respectively) and in the left lateral compared with sitting position (by 7.8%) (P < 0.05). Maternal stroke volume index, heart rate, and systolic blood pressure were higher in the lateral positions compared with the sitting and supine-tilt positions. We found no significant differences in fetal heart rate, pulsatility index, or resistivity index among positions. CONCLUSION: Positioning for neuraxial anesthesia may influence maternal hemodynamic variables. We found no difference in healthy fetal blood flow indices among positions, suggesting that these changes are not clinically significant. This study provides new physiological information on the changes that occur in a group in whom it has not been practical to study previously. Further study is necessary to determine whether these changes are significant in the presence of neuraxial anesthesia or in the high-risk parturient. PMID- 21642603 TI - Intravenous sodium bicarbonate verifies intravenous position of catheters in ventilated patients. AB - BACKGROUND: Extravasation is the unintentional injection or leakage of fluids into the perivascular or subcutaneous space resulting in potential tissue injury. In this 2-part prospective, controlled study, we assessed the safety of subcutaneously injected sodium bicarbonate in rats first. In the second part, the diagnostic utility of using IV diluted sodium bicarbonate to confirm placement of IV catheters in endotracheally intubated and ventilated rats and patients was tested. Diluted sodium bicarbonate was created using undiluted standard 8.4% (1 mEq/mL) sodium bicarbonate mixed in a 1:1 ratio with sterile water to achieve a final diluted concentration of 4.2% (0.5 mEq/mL). METHODS: Sodium bicarbonate (8.4% and 4.2%) was injected subcutaneously into 10 rats, and skin samples were evaluated. The hemodynamic and ventilatory effects of IV bicarbonate (2 mL/kg) in ventilated rats were measured. Subsequently, in 20 ASA physical status I and II mechanically ventilated patients, the effects of 50 mL of diluted 4.2% sodium bicarbonate or 0.9% normal saline, injected in a randomized order, were analyzed. RESULTS: Part 1: Undiluted (8.4%) subcutaneous sodium bicarbonate resulted in a small area of skin necrosis in 10% of skin samples (3 of 30) taken from rats. Minimal effects (mild scale crust and foci of regenerative epidermis beneath) were detected when a diluted solution was used. In ventilated rats, IV injection of diluted bicarbonate caused a significant increase in end-tidal carbon dioxide, whereas subcutaneous injection had no effect. In humans, diluted bicarbonate resulted in an end-tidal carbon dioxide increase (mean of 38 +/- 5 to 45 +/- 7 mm Hg) within 7 breaths. Injected normal saline did not result in any changes. Sodium bicarbonate was easily differentiated from normal saline injection by anesthesiologists observing the change in end-tidal carbon dioxide concentrations immediately after injection. CONCLUSION: The injection of diluted sodium bicarbonate (in mechanically ventilated patients) can be used to reliably identify the correct location of an IV catheter by an increase in the exhaled carbon dioxide concentration. Although we found no skin damage with 4.2% (0.5 mEq/mL) sodium bicarbonate, safety and efficacy should be further evaluated in future studies. PMID- 21642604 TI - Is ultrasound guidance advantageous for interventional pain management? A review of acute pain outcomes. AB - BACKGROUND: Ultrasound (US) guidance for peripheral nerve blockade has gained popularity worldwide. The reported benefits of real-time sonographic visualization compared with traditional nerve localization techniques generally apply to procedural and technical block-related outcomes whereas acute pain related outcomes are featured less prominently. In this review, we evaluated the effect of US guidance compared with traditional nerve localization techniques for interventional management of acute pain and acute pain-related outcomes. METHODS: We performed a systematic search of MEDLINE, EMBASE, and the Cochrane Central Register of Controlled Clinical Trials (from January 1990 to January 2011) to identify randomized controlled trials evaluating the effects of US guidance on acute pain and related outcomes compared with traditional nerve localization techniques. Studies were excluded if they did not report at least one of the following acute pain outcomes: pain severity, opioid consumption, sensory block duration, and time to first analgesic request. Related outcomes were classified as follows: patient related (opioid-related adverse effects, patient satisfaction, postoperative cognitive deficit); anesthesia related (unwanted motor block, perineural catheter failure, morbidity, development of chronic pain); surgery related (hospital readmission, ability to ambulate); and hospital related (length of stay, cost). Promising novel applications of US guidance for acute pain management were also sought for discussion purposes. RESULTS: We identified 23 randomized controlled trials, including 1674 patients, that compared US guidance with and without peripheral nerve stimulation with peripheral nerve stimulation alone or anatomical landmark techniques. Of the 16 studies that evaluated pain severity, 8 reported improvement with US guidance; however, only 1 study reported a difference between US guidance and the comparator of >1 interval on the numeric rating pain scale. Eight studies evaluated sensory block duration and 3 of these reported prolonged block duration with US guidance. Seven studies evaluated opioid consumption, of which 3 reported a reduction with US guidance. Three studies evaluated time to first analgesic request, of which 2 favored US guidance. We uncovered no significant differences between US guidance and traditional nerve localization techniques for any other related outcome. US guidance was not found to be inferior compared with traditional nerve localization techniques for any outcome. Nonrandomized data suggest that US-guided transversus abdominis plane blocks may offer analgesic benefit over standard analgesic therapy, but has not been compared with an anatomical landmark technique. CONCLUSIONS: At present, there is insufficient evidence in the contemporary literature to define the effect of US guidance on acute pain and related outcomes compared with traditional nerve localization techniques for interventional acute pain management. PMID- 21642605 TI - Pharmacokinetics and tissue penetration of cefoxitin in obesity: implications for risk of surgical site infection. AB - BACKGROUND: Obesity is a significant risk factor for surgical site infections (SSIs), for poorly understood reasons. SSIs are a major cause of morbidity, prolonged hospitalization, and increased health care cost. Drug disposition in general is frequently altered in the obese. Preoperative antibiotic administration, achieving adequate tissue concentrations at the time of incision, is an essential strategy to prevent SSIs. Nonetheless, there is little information regarding antibiotic concentrations in obese surgical patients. This investigation tested the hypothesis that the prophylactic antibiotic cefoxitin may have delayed and/or diminished tissue penetration in the obese. METHODS: Plasma and tissue concentrations of cefoxitin were determined in obese patients undergoing abdominal and pelvic surgery (body mass index 43 +/- 10 kg/m(2), n = 14, 2 g cefoxitin) and in normal-weight patients and healthy volunteers (body mass index 20 +/- 2 kg/m(2), n = 13, 1 g cefoxitin). Tissue concentrations were measured using a microdialysis probe in the subcutaneous layer of the abdomen, and in adipose tissue excised at the time of incision and wound closure. RESULTS: Plasma concentrations and area under the concentration-time curve (AUC) were approximately 2-fold higher in the obese patients because of the 2-fold-higher dose. Dose-normalized concentrations were higher, although AUCs were not significantly different. Measured and dose-normalized subcutaneous cefoxitin concentrations and AUCs in the obese patients were significantly lower than in the normal-weight subjects. There was an inverse relationship between cefoxitin tissue penetration (AUC(tissue)/AUC(plasma) ratio) and body mass index. Tissue penetration was substantially lower in the obese patients (0.08 +/- 0.07 vs 0.37 +/- 0.26, P < 0.05). Adipose tissue cefoxitin concentrations in obese patients were only 7.8 +/- 7.3 and 2.7 +/- 1.4 MUg/g, respectively, at incision and closure, below the minimum inhibitory concentration of 8 and 16 MUg/mL, respectively, for aerobic and anaerobic microorganisms. CONCLUSION: Obese surgical patients have impaired tissue penetration of the prophylactic antibiotic cefoxitin, and inadequate tissue concentrations despite increased clinical dose (2 g). Inadequate tissue antibiotic concentrations may be a factor in the increased risk of SSIs in obese surgical patients. Additional studies are needed to define doses achieving adequate tissue concentrations. PMID- 21642606 TI - The ability of pulse pressure variations obtained with CNAPTM device to predict fluid responsiveness in the operating room. AB - BACKGROUND: Respiratory-induced pulse pressure variations obtained with an arterial line (DeltaPP(ART)) indicate fluid responsiveness in mechanically ventilated patients. The Infinity(r) CNAPTM SmartPod(r) (Drager Medical AG & Co. KG, Lubeck, Germany) provides noninvasive continuous beat-to-beat arterial blood pressure measurements and a near real-time pressure waveform. We hypothesized that respiratory-induced pulse pressure variations obtained with the CNAP system (DeltaPP(CNAP)) predict fluid responsiveness as well as DeltaPP(ART) predicts fluid responsiveness in mechanically ventilated patients during general anesthesia. METHODS: Thirty-five patients undergoing vascular surgery were studied after induction of general anesthesia. Stroke volume (SV) measured with the VigileoTM/FloTracTM (Edwards Lifesciences, Irvine, CA), DeltaPP(ART), and DeltaPP(CNAP) were recorded before and after intravascular volume expansion (VE) (500 mL of 6% hydroxyethyl starch 130/0.4). Subjects were defined as responders if SV increased by >=15% after VE. RESULTS: Twenty patients responded to VE and 15 did not. The correlation coefficient between DeltaPP(ART) and DeltaPP(CNAP) before VE was r = 0.90 (95% confidence interval [CI] = 0.84-0.96; P < 0.0001). Before VE, DeltaPP(ART) and DeltaPP(CNAP) were significantly higher in responders than in nonresponders (P < 0.0001). The values of DeltaPP(ART) and DeltaPP(CNAP) before VE were significantly correlated with the percent increase in SV induced by VE (respectively, r(2) = 0.50; P < 0.0001 and r(2) = 0.57; P < 0.0001). Before VE, a DeltaPP(ART) >10% discriminated between responders and nonresponders with a sensitivity of 90% (95% CI = 69%-99%) and a specificity of 87% (95% CI = 60% 98%). The area under the receiver operating characteristic (ROC) curve was 0.957 +/- 0.035 for DeltaPP(ART). Before VE, a DeltaPP(CNAP) >11% discriminated between responders and nonresponders with a sensitivity of 85% (95% CI = 62%-97%) and a specificity of 100% (95% CI = 78%-100%). The area under the ROC curve was 0.942 +/- 0.040 for DeltaPP(CNAP). There was no significant difference between the area under the ROC curve for DeltaPP(ART) and DeltaPP(CNAP). CONCLUSIONS: A value of DeltaPP(CNAP) >11% has a sensitivity of at least 62% in predicting preload dependent responders to VE in mechanically ventilated patients during general anesthesia. PMID- 21642607 TI - The disclosure of unanticipated outcomes of care and medical errors: what does this mean for anesthesiologists? AB - The disclosure of unanticipated outcomes to patients, including medical errors, has received considerable attention of late. The discipline of anesthesiology is a leader in patient safety, and as the doctrine of full disclosure gains momentum, anesthesiologists must become acquainted with these philosophies and practices. Effective disclosure can improve doctor-patient relations, facilitate better understanding of systems, and potentially decrease medical malpractice costs. However, many physicians remain wary of discussing errors with patients due to concern about litigation, the communication challenges of disclosure, and loss of self-esteem. As a result, harmful errors are often not disclosed to patients. Disclosure poses special challenges for anesthesiologists. There is often very limited time before the anesthetic in which to build the patient physician relationship, and anesthesiologists usually function within complex health care teams. Other team members such as the surgeon may have different perspectives on what the patient should be told about operating room errors. The anesthesiologist may still be physically caring for the patient while the surgeon has the initial discussion with the family about the event. As a result the anesthesiologist may be excluded from the planning or conduct of the important initial disclosure conversations. New disclosure strategies are needed to engage anesthesiologists as active participants in the disclosure of unanticipated outcomes. Anesthesiologists should be aware of the emerging best practices surrounding disclosure, as well as the training opportunities and disclosure support resources that are increasingly available. Innovative models should be developed that promote collaboration between all perioperative team members in the disclosure process. There are important opportunities for anesthesiologists to play a leading role in defining specialty-specific disclosure practices and to more effectively meet patients' needs for disclosure after unanticipated outcomes and medical errors. PMID- 21642608 TI - Preoperative arterial pulse pressure has no apparent association with perioperative mortality after lower extremity arterial bypass. AB - BACKGROUND: Arterial pulse pressure hypertension is associated with perioperative morbidity and mortality in cardiac surgery patients. However, its association with perioperative mortality in other high-risk surgical populations has not been determined. In this study, we tested the hypothesis that increased preoperative arterial pulse pressure is associated with 30-day and 1-year all-cause mortality after lower extremity arterial bypass surgery. METHODS: A retrospective review of patients who had infrainguinal arterial bypass surgery at a single center over a 6-year period (January 2002 to January 2008) was performed (n = 556). Mean, systolic, and diastolic arterial blood pressure were determined from a single noninvasive oscillometric blood pressure cuff reading in the operating room before the administration of anesthetic drugs. Pulse pressure was calculated from this measurement in a retrospective manner by subtracting diastolic pressure from systolic pressure. Mortality for all subjects was determined using the social security death index. Comorbid conditions, preoperative medications, and anesthetic techniques were recorded. Univariate and multivariate analyses were performed to evaluate the association between arterial pulse pressure and the primary outcome variables, and all-cause 30-day and 1-year mortality. RESULTS: Of the 556 patients, a large percentage had elevated pulse pressure (44.9% had pulse pressure >=80). Thirty-day mortality was 5.1% and 1-year mortality was 17.8%. There was no apparent association between preoperative pulse pressure and 30-day (P = 0.35) or 1-year (P = 0.14) all-cause mortality. Independent predictors of 30 day mortality were age >=80 years (P = 0.02), ASA physical status >=IV (P = 0.04), baseline creatinine >2.0 mg/dL (P < 0.0001), and emergency surgery (P = 0.009). The same variables were associated with 1-year mortality, as were the Lee's Revised Cardiac Risk Index score, female gender, and gangrene or ulcer as an indication for surgery. CONCLUSION: Our results suggest that increased preoperative arterial pulse pressure might not be associated with all-cause mortality after lower extremity arterial bypass surgery. PMID- 21642609 TI - Upregulation of DeltaFosB by propofol in rat nucleus accumbens. AB - BACKGROUND: It is well established that all drugs of abuse converge onto common circuitry and induce chronic addiction by modulating the addictive signaling molecules such as DeltaFosB in the mesocorticolimbic system. Recent case reports suggest that propofol may have abuse potential. However, there is no direct evidence showing that propofol has an effect on the key addictive signaling molecules in the mesocorticolimbic system. In this study, we determined the effect of propofol on the expression of DeltaFosB in rat nucleus accumbens (NAc) and the potential mechanism involved. METHODS: To determine the effect of propofol on the expression of DeltaFosB in rat NAc, 2 well-known addictive agents, ethanol and nicotine, were used as positive controls. Experiments were conducted on 36 male Sprague-Dawley rats (150 to 200 g). These animals were divided into 4 treatment groups: vehicle (saline), propofol (10 mg/kg), ethanol (1 g/kg), and nicotine (0.5 mg/kg). All drugs were administered by intraperitoneal injection twice per day for 7 days. The animals were then killed and their NAc were isolated for DeltaFosB measurements. RESULTS: As expected, both ethanol and nicotine significantly increased DeltaFosB expression. Intriguingly, propofol elicited a robust increase in DeltaFosB expression similar to that of ethanol and nicotine. Moreover, the dopamine receptor D1, an upstream molecule of DeltaFosB, was also significantly upregulated by propofol. CONCLUSIONS: In the current study, we have identified, for the first time, that propofol is able to induce the addictive signaling molecule DeltaFosB in NAc via dopamine receptor D1. This new evidence at the molecular level suggests that propofol may have abuse potential. PMID- 21642610 TI - Prior epidural lidocaine alters the pharmacokinetics and drug effects of extended release epidural morphine (DepoDur(r)) after cesarean delivery. AB - BACKGROUND: A potential physicochemical interaction between epidural local anesthetics and extended-release epidural morphine (EREM) could negate the sustained release. In this study, we sought to determine the pharmacokinetic and drug effects of prior epidural lidocaine administration on EREM. METHODS: Thirty healthy women undergoing cesarean delivery were enrolled in this randomized study. Patients received 8 mg EREM 1 hour after either a combined spinal-epidural (intrathecal bupivacaine and fentanyl 20 MUg with no epidural medication; group SE) or an epidural anesthetic (epidural 2% lidocaine with fentanyl 100 MUg; group E). Maximal concentration (Cmax), time to Cmax (Tmax), and AUC(0-last) (area under the concentration-time curve until the last plasma concentration that was below the limit of quantitation) for morphine levels were determined from a plasma sample at 0, 5, 10, 15, and 30 minutes, and 1, 4, 8, 12, 24, 36, 48, and 72 hours. Drug effects including pain, analgesic use, and side effects were measured for 72 hours after cesarean delivery. RESULTS: Epidural lidocaine administration (20-35 mL) 1 hour before epidural EREM administration increased the Cmax in group E (11.1 +/- 4.9) compared with group SE (8.3 +/- 7.1 ng/mL) (P = 0.038). There were no significant effects on Tmax and AUC(0-last) of venous morphine between the groups (P > 0.05). There was an increased incidence in vomiting, oxygen use, and hypotension in group E (patients who received lidocaine before EREM). CONCLUSION: A large dose of epidural lidocaine 1 hour before EREM administration alters the pharmacokinetics and drug effects of EREM. Clinicians must apply caution when EREM is administered even 1 hour after an epidural lidocaine "top-up" for cesarean delivery. PMID- 21642611 TI - An in vitro investigation of the coagulation effects of exogenous oxytocin using thromboelastography in healthy parturients. AB - BACKGROUND: We investigated the coagulation effects in vitro of exogenous oxytocin in whole blood of healthy term parturients. METHODS: Thromboelastography (TEG(r)) was performed on kaolin-activated citrated blood samples from 25 healthy, term, nonlaboring parturients. We compared the in vitro effects on the maternal thromboelastographic profile of 3 different exogenous oxytocin concentrations (22.5, 30.1, and 32.9 MUU/mL) and a control (0 MUU/mL). These exogenous oxytocin concentrations were chosen to approximate maternal plasma oxytocin concentrations during elective cesarean delivery, vaginal delivery, and nonelective cesarean delivery, respectively. RESULTS: Increasing the oxytocin concentration was significantly associated with hypercoagulable effects as observed with TEG(r) (decreasing reaction time, clot formation time, and Tmax; increasing alpha angle and maximum rate of thrombus generation). Compared with control samples, the median percentage change (interquartile range) in TEG(r) values for samples with the highest exogenous oxytocin concentration (32.9 MUU/mL) was largest for reaction time: -40.3% (-45.8%, -22.2%); and Tmax: -39.2% (-42.9%, -28.5%). CONCLUSION: The results of this in vitro investigation suggest that exogenous oxytocin is associated with modest hypercoagulable effects in the maternal blood of healthy term parturients. PMID- 21642612 TI - Ross C. Terrell, PhD, an anesthetic pioneer. AB - On December 30, 2010, Ross C. Terrell, PhD, died. With his passing at age 85, we lost one of the pioneers of modern anesthesia. Terrell synthesized most of the inhalation anesthetics used today, including desflurane, enflurane, isoflurane, and sevoflurane. PMID- 21642613 TI - The median effective dose of ketamine and gabapentin in opioid-induced hyperalgesia in rats: an isobolographic analysis of their interaction. AB - BACKGROUND: Ketamine and gabapentin have been shown to prevent the delayed hyperalgesia induced by short-term use of systemic opioids. The mechanism of this action is believed to be likely at the spinal level, through an antagonism of the N-methyl-D-aspartate receptors for ketamine, and through a specific binding site for gabapentin. In this study, we sought to determine the nature of the interaction of these 2 mechanistically distinct antihyperalgesic drugs in a model of opioid-induced hyperalgesia in rats. The median effective antihyperalgesic doses of each drug and of their combination were first defined, to assess the nature of the interaction using an isobolographic analysis. METHODS: Long-lasting hyperalgesia was induced in male Sprague Dawley rats with subcutaneous fentanyl (4 injections, 60 MUg/kg per injection at 15-minute intervals) resulting in a total dose of 240 MUg/kg. Subcutaneous ketamine, or intraperitoneal gabapentin, or their combination was administered 30 minutes before the first subcutaneous fentanyl injection. Sensitivity to nociceptive stimuli (von Frey filaments) was assessed on the day of the experiment and on the day after injections. The dose of ketamine and gabapentin received by a particular animal was determined by the response of the previous animal of the same group, using an up-and-down technique. Initial doses were 10 mg/kg and 300 mg/kg, with dose adjustment intervals of 1 mg/kg and 30 mg/kg, in the ketamine and gabapentin groups, respectively. The initial doses of ketamine and gabapentin were 5 mg/kg and 150 mg/kg, respectively, in the ketamine-gabapentin group, with the same dose adjustment intervals. Antihyperalgesic efficacy was defined as complete prevention of hyperalgesia on the day after drug injections. RESULTS: The median effective antihyperalgesic doses (median value and 95% confidence interval) of ketamine and gabapentin were 12.4 mg/kg (11.7-13.1 mg/kg) and 296.3 mg/kg (283.5 309.1 mg/kg), respectively. The median effective antihyperalgesic dose of the combination was 4.3 mg/kg (3.7-4.6 mg/kg) for ketamine and 123.9 mg/kg (111.1 136.7 mg/kg) for gabapentin. CONCLUSION: The isobolographic analysis demonstrated that the combination of the 2 drugs produces effective antihyperalgesia with a supraadditive (synergistic) action. PMID- 21642614 TI - Comparison of mounting methods for the evaluation of fibers by phase contrast microscopy. AB - The objectives of this study were to evaluate mounting methods for fiber examination of air sample filters by phase contrast microscopy (PCM) and to evaluate differences in fiber counts that might be due to fiber movement. Acetone/triacetin (AT) with various amounts of triacetin and acetone/Euparal (AE) where the mounting medium was placed between the cleared filter wedge and the coverslip were tested as a function of time. Field sample slides collected from a taconite iron-ore processing mill, a tremolitic talc-ore processing mill, and from around a crusher in a meta-basalt stone quarry were prepared with relocatable coverslips to revisit the same field areas on the slides. For each slide, three or four field areas were randomly selected and pictures were taken every 2 weeks to determine any sign of fiber movement over time. For 11 AT slides (named as AT-3.5) prepared with 3.5 MUl of the mounting medium according to the NIOSH 7400 method, no fiber movements were detected over 59 weeks. On the other hand, AT slides prepared with larger quantities (10, 15, and 20 MUl) of the mounting medium (named as AT-10) and AE slides prepared with ~10 MUl mounting medium showed fiber movement from the eighth day at the earliest. Fiber movement began earlier for the slides mounted with excess triacetin than for those mounted with Euparal. The sample slide storage method, either vertically or horizontally, did not seem to accelerate fiber movement. Additionally, two other modified methods, dimethylformamide solution/Euparal (mDE) and dimethylformamide solution/triacetin (mDT), were also prepared where the mounting medium was placed between the cleared filter wedge and the glass slide. The findings of fiber movements were similar; when 3.5 MUl of triacetin was used for the mDT slides, fiber movements were not detected, while fibers on slides prepared with 10 MUl triacetin (mDT-10) moved around. No fiber movements were observed for the mDE slides at any time during 59 weeks. Once fiber movement started, fibers moved over distances measured from 4 MUm and up to >1000 MUm within 22 weeks. However, since then, no further fiber movements have been observed in any field sample slides. Additional sample slides, two Amosite and two chrysotile, were prepared from Union for International Cancer Control (UICC) samples using the AT method with 5 MUl triacetin mounting medium. Fiber movements were also observed in these samples; chrysotile fibers began to migrate in 3 weeks, while Amosite fiber movement started after 3 months. Although fiber movement was observed for the AT 10, AE, and mDT-10 sample slides, fiber counts were not significantly different from AT-3.5 and mDE samples that exhibited no fiber movement. Although fiber counts would not be significantly changed by fiber movement, the type and amount of mounting medium for sample slide preparation remains critical for issues such as quality assurance and training of analysts by revisiting the same fibers. PMID- 21642615 TI - The influence of valproic acid and carbamazepine treatment on serum biotin and zinc levels and on biotinidase activity. AB - We determined the serum concentration of biotin, zinc, antiepileptic drugs, and biotinidase enzyme activity in 20 children treated with valproic acid, in 10 children treated with carbamazepine, and in 75 age- and sex-matched healthy controls. There were no significant differences in the serum levels of biotin, and biotinidase enzyme activity between the patients treated with valproic acid, the patients treated with carbamazepine, and the control group. Zinc serum levels were lower in the patients treated with valproic acid and with carbamazepine than in the control group, but within the normal range. Hair loss was observed in 3 patients treated with valproic acid, with normal serum levels of biotin, zinc, and biotinidase activity, and the alopecia disappeared with the oral administration of biotin (10 mg/d) in 3 months. These results suggest that the treatment with valproic acid does not alter the serum levels of biotin, zinc, and biotinidase enzyme activity. PMID- 21642616 TI - Quantitative comparison between in vivo DNA adduct formation from exposure to selected DNA-reactive carcinogens, natural background levels of DNA adduct formation and tumour incidence in rodent bioassays. AB - This study aimed at quantitatively comparing the occurrence/formation of DNA adducts with the carcinogenicity induced by a selection of DNA-reactive genotoxic carcinogens. Contrary to previous efforts, we used a very uniform set of data, limited to in vivo rat liver studies in order to investigate whether a correlation can be obtained, using a benchmark dose (BMD) approach. Dose-response data on both carcinogenicity and in vivo DNA adduct formation were available for six compounds, i.e. 2-acetylaminofluorene, aflatoxin B1, methyleugenol, safrole, 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline and tamoxifen. BMD(10) values for liver carcinogenicity were calculated using the US Environmental Protection Agency BMD software. DNA adduct levels at this dose were extrapolated assuming linearity of the DNA adduct dose response. In addition, the levels of DNA adducts at the BMD(10) were compared to available data on endogenous background DNA damage in the target organ. Although for an individual carcinogen the tumour response increases when adduct levels increase, our results demonstrate that when comparing different carcinogens, no quantitative correlation exists between the level of DNA adduct formation and carcinogenicity. These data confirm that the quantity of DNA adducts formed by a DNA-reactive compound is not a carcinogenicity predictor but that other factors such as type of adduct and mutagenic potential may be equally relevant. Moreover, comparison to background DNA damage supports the notion that the mere occurrence of DNA adducts above or below the level of endogenous DNA damage is neither correlated to development of cancer. These data strongly emphasise the need to apply the mode of action framework to understand the contribution of other biological effect markers playing a role in carcinogenicity. PMID- 21642617 TI - Aristolochic acid-induced carcinogenesis examined by ACB-PCR quantification of H Ras and K-Ras mutant fraction. AB - Aristolochic acid (AA) is a strong cytotoxic nephrotoxin and carcinogen associated with the development of urothelial cancer in humans. AA induces forestomach, kidney and urothelial tract tumours in rats and mice. This study was conducted to characterise AA's carcinogenic mechanism of action and compare allele-specific competitive blocker-polymerase chain reaction (ACB-PCR)-based early detection of carcinogenic effect using two different tumour-relevant endpoints. H-Ras codon 61 CAA->CTA mutation was analysed because it is found in rodent forestomach tumours and A:T->T:A transversion is the predominant mutational specificity induced by AA. K-Ras codon 12 GGT->GAT mutation was analysed because it is a common spontaneous mutation present in various rodent tissues and may be a useful generic biomarker for carcinogenic effect. DNA samples from Big Blue rats treated with 0, 0.1, 1.0 or 10.0 mg AA/kg body weight (bw) by gavage, 5 days/week for 12 weeks were used in ACB-PCR in order to examine the induction of the two specific mutations. A significant dose-dependent induction of H-Ras mutant fraction (MF) was observed in liver and kidney. Statistically significant correlations were observed between AA-induced DNA adduct levels or cII mutant frequencies (previously measured in the same rats) and H-Ras MF measurements. No correlation between AA dose and K-Ras MF was found in liver or kidney, although there was a significant induction of K-Ras mutation in kidneys exposed to 0.1 mg/kg bw AA relative to controls. Thus, the data establish a straightforward dose-related increase in H-Ras MF due to fixation of AA-induced DNA adducts, whereas the common spontaneous K-Ras mutation showed a non-monotonic dose-response, consistent with loss of non-targeted mutation at cytotoxic doses. PMID- 21642618 TI - Aquaporin-0 interacts with the FERM domain of ezrin/radixin/moesin proteins in the ocular lens. AB - PURPOSE: Aquaporin 0 (AQP0) is the major intrinsic protein in the lens and is essential for establishing proper fiber cell structure and organization. Cytoskeletal proteins that directly interact with the C terminus of AQP0 are identified herein. METHODS: The water-insoluble fraction of lens fiber cells was chemically cross-linked, and cross-linked peptides with the C terminus of AQP0 were identified by mass spectrometry. Coimmunoprecipitation and AQP0 C-terminal peptide pulldown experiments were used to confirm the protein-protein interaction. RESULTS: Unexpectedly, AQP0 was found to directly associate with ezrin/radixin/moesin (ERM) family members, proteins that are involved in linkage of actin filaments to the plasma membrane. Cross-linked peptides were detected between AQP0 and degenerate sequences of ezrin and radixin; however, AQP0 interaction with ezrin is believed to play a more significant function in the lens because of its higher level of expression and observed ezrin-specific cross linking. The interaction was found to occur between the C terminus of AQP0 and subdomains F1 and F3 of ERM proteins. The interaction between AQP0 and ezrin was confirmed by coimmunoprecipitation and AQP0 C-terminal peptide pulldown experiments. CONCLUSIONS: Considering the important known functions of the cellular actin cytoskeleton in fiber cell differentiation, the interaction of AQP0 and ERM proteins may play an important role in fiber cell morphology, elongation, and organization. PMID- 21642619 TI - Homocysteine-mediated modulation of mitochondrial dynamics in retinal ganglion cells. AB - PURPOSE: To evaluate the effect of excess homocysteine on the regulation of retinal ganglion cell mitochondrial dynamics. METHODS: Mice deficient in cystathionine-beta-synthase (cbs) were used as a model of hyperhomocysteinemia. Gene and protein expression analyses of Opa1 and Fis1 were performed on cbs+/- neural retinas. Mitochondria within retinal ganglion cell axons underwent systematic ultrastructural analysis to measure area, length, width, and the distance between the mitochondria and the axon wall. Primary mouse ganglion cells were cultured, treated with homocysteine, and assessed for levels of Opa1 and Fis1 protein, the number of mitochondria per length of neurite, and levels of cleaved caspase-3. RESULTS: Opa1 and Fis1 protein levels in cbs+/- neural retinas were elevated to 191.00% +/- 26.40% and 226.20% +/- 4.57%, respectively, compared with wild-type. Mitochondria of cbs+/- retinas were smaller in all parameters studied, including area (0.32 +/- 0.01 MUm2 vs. 0.42 +/- 0.02 MUm2), compared with wild-type. Primary ganglion cells treated with homocysteine had elevations in Opa1 and Fis1 proteins, a significantly higher number of mitochondria per length of neurite (0.1781 +/- 0.017 vs. 0.1156 +/- 0.012), and significantly higher levels of cleaved caspase-3 compared with control. CONCLUSIONS: This study provides the first evidence that homocysteine-induced ganglion cell loss involves the dysregulation of mitochondrial dynamics, both in vivo and in vitro. The present data suggest increased mitochondrial fission as a novel mechanism of homocysteine toxicity to neurons. Of particular relevance are glaucoma and Alzheimer's disease, neurodegenerative diseases that are associated with hyperhomocysteinemia and, more recently, have implicated increased mitochondrial fission in their pathogeneses. PMID- 21642620 TI - High-resolution photoreceptor imaging in idiopathic macular telangiectasia type 2 using adaptive optics scanning laser ophthalmoscopy. AB - PURPOSE: To study pathologic changes in the photoreceptors in eyes with idiopathic macular telangiectasia (MacTel) type 2 using adaptive optics scanning laser ophthalmoscopy (AO-SLO). METHODS: Thirteen eyes with nonproliferative MacTel type 2 and 10 normal eyes underwent a full ophthalmologic examination, spectral-domain optical coherence tomography (SD-OCT), and imaging with an original prototype AO-SLO system. All eyes with MacTel type 2 were examined with fluorescein angiography (FA), fundus autofluorescence (FAF), confocal blue reflectance (CBR), and fundus-monitoring microperimetry (MP). RESULTS: All eyes with MacTel type 2 had ring-like dark areas and/or small patchy regions on AO-SLO images; significantly lower cone density than that of normal eyes in each hemisphere at 0.5 mm from the foveal center; an area with parafoveal reflectance in CBR that was larger than the hyperfluorescence area in FA, the area of increased FAF, the dark areas on AO-SLO, and the area of decreased retinal sensitivity on MP. Dark areas on AO-SLO roughly corresponded to the leakage area in FA, but dark areas were also seen in areas without FA leakage in 11 eyes, including an eye with the earliest clinical signs of MacTel. Visual acuity and retinal sensitivity correlated with mean cone density 0.5 mm from the center of the fovea. CONCLUSIONS: In eyes with MacTel type 2, AO-SLO revealed unique dark regions in the cone mosaic and decreased cone density that was associated with decreased vision, even in areas with normal vasculature, which suggests that this feature represents early neuronal changes involved in the pathogenesis of MacTel type 2. PMID- 21642621 TI - Expression of PDGFRalpha is a determinant of the PVR potential of ARPE19 cells. AB - PURPOSE: Previous studies indicate that the expression of platelet-derived growth factor (PDGF) receptor alpha (PDGFRalpha) dramatically increases the ability of fibroblasts to induce experimental proliferative vitreoretinopathy (PVR). The purpose of this study was to determine whether PDGFRalpha contributed to the PVR potential of retinal pigment epithelial (RPE) cells, one of the most abundant cell types in PVR membranes. METHODS: PDGFRalpha expression in human ARPE19 cells was increased or decreased by stably expressing the PDGFRalpha cDNA or short hairpin (sh) RNA directed at PDGFRalpha, respectively. The level of PDGFRalpha expression in the resulting panel of cell lines was either barely detectable (KD), standard (similar to the level of primary RPE cells), or overexpressed approximately 80-fold. Western blot analysis was used to assess the level of p53 and the activation state of PDGFRalpha and Akt. The following cellular responses were monitored: proliferation, apoptosis, and contraction. The PVR potential of cells was tested in a rabbit model of PVR in which cells were coinjected with platelet-rich plasma into the vitreous. RESULTS: Comparison of KD and overexpressing cells indicated that high-level expression of PDGFRalpha dramatically augmented signaling events, cellular responses, and the PVR potential of ARPE19 cells. However, all these outcomes were also significantly increased, albeit not as robustly, by PDGFRalpha expression to the level typically present in RPE cells. CONCLUSIONS: Even though RPE cells express substantially less PDGFRalpha than fibroblasts, it significantly boosts PVR related signaling events, cellular responses, and the PVR potential of ARPE19 cells. These studies suggest that inhibiting activation, signaling, or both by PDGFRalpha has the potential to prevent the development of PVR. PMID- 21642622 TI - Role of TGFbeta/Smad signaling in gremlin induction of human trabecular meshwork extracellular matrix proteins. AB - PURPOSE: The bone morphogenic protein (BMP) antagonist gremlin is elevated in glaucomatous trabecular meshwork (TM) cells and tissues and elevates intraocular pressure (IOP). Gremlin also blocks BMP4 inhibition of transforming growth factor (TGF)-beta2 induction of TM extracellular matrix (ECM) proteins. The purpose of this study was to determine whether Gremlin regulates ECM proteins in cultured human TM cells. METHODS: Human TM cells were treated with recombinant gremlin to determine the effects on ECM gene and protein expression. Expression of the ECM genes FN, COL1, PAI1, and ELN was examined in cultured human TM cells by quantitative RT-PCR and Western immunoblot analysis. TM cells were pretreated with TGFBR inhibitors (LY364947, SB431542 or TGFBR1/TGFB2 siRNAs), inhibitors of the Smad signaling pathway (SIS3 or Smad2/3/4 siRNAs), or CTGF siRNA to identify the signaling pathway(s) involved in gremlin induction of ECM gene and protein expression. RESULTS: All ECM genes analyzed (FN, COL1, PAI1, and ELN) were induced by gremlin. This gremlin induction of ECM genes and protein expression was blocked by inhibitors of TGFBR and the canonical Smad2/3/4 and CTGF signaling pathways. CONCLUSIONS: Gremlin employs canonical TGFbeta2/Smad signaling to induce ECM genes and proteins in cultured human TM cells. Gremlin also induces both TGFbeta2 and CTGF, which can act downstream to mediate some of these ECM changes in TM cells. PMID- 21642624 TI - Rapid, high-accuracy detection of strabismus and amblyopia using the pediatric vision scanner. AB - Purpose. The Pediatric Vision Scanner (PVS) detects strabismus by identifying ocular fixation in both eyes simultaneously. This study was undertaken to assess the ability of the PVS to identify patients with amblyopia or strabismus, particularly anisometropic amblyopia with no measurable strabismus. Methods. The PVS test, administered from 40 cm and requiring 2.5 seconds of attention, generated a binocularity score (BIN, 0%-100%). We tested 154 patients and 48 controls between the ages of 2 and 18 years. BIN scores of amblyopic children and controls were measured, and 21 children received sequential PVS measurements to detect any changes in BIN resulting from amblyopia treatment. Results. With the pass/refer threshold set at BIN 60%, sensitivity and specificity were 96% for the detection of amblyopia or strabismus. Assuming a 5% prevalence of amblyopia or strabismus, the inferred positive and negative predictive values of the PVS were 56% and 100%, respectively. Fixation accuracy was significantly reduced in amblyopic eyes. In anisometropic amblyopia patients treated successfully, the BIN improved to 100%. Conclusions. The PVS identified children with amblyopia or strabismus with high sensitivity and specificity, while successful treatment restored normal BIN scores in amblyopic patients without strabismus. The results support the hypothesis that the PVS detects strabismus and amblyopia directly. Future strategies for screening by nonspecialists may thus be based on diagnostic detection of amblyopia and strabismus rather than the estimation of risk factors, allowing for rapid, accurate identification of children with amblyopia early in life when it is most amenable to treatment. PMID- 21642623 TI - Image defocus and altered retinal gene expression in chick: clues to the pathogenesis of ametropia. AB - PURPOSE: Because of the retina's role in refractive development, this study was conducted to analyze the retinal transcriptome in chicks wearing a spectacle lens, a well-established means of inducing refractive errors, to identify gene expression alterations and to develop novel mechanistic hypotheses about refractive development. METHODS: One-week-old white Leghorn chicks wore a unilateral spectacle lens of +15 or -15 D for 6 hours or 3 days. With total RNA from the retina/(retinal pigment epithelium, RPE), chicken gene microarrays were used to compare gene expression levels between lens-wearing and contralateral control eyes (n = 6 chicks for each condition). Normalized microarray signal intensities were evaluated by analysis of variance, using a false discovery rate of <10% as the statistical criterion. Selected differentially expressed genes were validated by qPCR. RESULTS: Very few retina/RPE transcripts were differentially expressed after plus lens wear. In contrast, approximately 1300 transcripts were differentially expressed under each of the minus lens conditions, with minimal overlap. For each condition, low fold-changes typified the altered transcriptome. Differentially regulated genes under the minus lens conditions included many potentially informative signaling molecules and genes whose protein products have roles in intrinsic retinal circadian rhythms. CONCLUSIONS: Plus or minus lens wear induce markedly different, not opposite, alterations in retina/RPE gene expression. The initial retinal responses to defocus are quite different from those when the eye growth patterns are well established, suggesting that different mechanisms govern the initiation and persistence or progression of refractive errors. The gene lists identify promising signaling candidates and regulatory pathways for future study, including a potential role for circadian rhythms in refractive development. PMID- 21642625 TI - Associations of complement factor H and smoking with early age-related macular degeneration: the ALIENOR study. AB - PURPOSE: To assess the associations of complement factor H (CFH) Y402H polymorphism and smoking with specific features of early AMD (type, location, and area). METHODS: The ALIENOR study is a population-based study of age-related eye diseases in 963 residents of Bordeaux (France), aged 73 years or more. AMD features were graded from nonmydriatic color retinal photographs. CFH Y402H was genotyped by using DNA extracted from blood. Statistical analyses included 796 subjects with complete data. RESULTS: CFH CC genotype was strongly associated with late neovascular AMD (OR, 6.0; 95% confidence interval [CI], 1.5-23.5) but not with late atrophic AMD (OR, 0.9; 95% CI, 0.2-4.3). Among early characteristics, it was associated with central soft drusen (within 500 MUm of the fovea), whether of intermediate (63-125 MUm; OR, 2.7; 95% CI, 1.5-4.8), or large (>125 MUm; OR, 5.9; 95% CI, 2.2-15.7) size, but not with pericentral soft drusen (500-3000 MUm from the fovea). It was also strongly associated with a large central area of soft drusen (OR, 5.7; 95% CI, 1.7-19.2). Similarly, heavy smoking (>20 pack-years) was strongly associated with central large drusen (OR, 3.9; 95% CI, 1.6-9.6) and a large central area of drusen (OR, 3.5; 95% CI, 1.2 10.0), but not with pericentral soft drusen. By contrast, both CFH CC and smoking tended to be more strongly associated with pericentral pigmentary abnormalities. CONCLUSIONS: Location of abnormalities, together with type and area, may prove useful for the identification of subjects at high risk for late AMD. PMID- 21642626 TI - Significant correlation between electroretinogram parameters and ocular vascular endothelial growth factor concentration in central retinal vein occlusion eyes. AB - PURPOSE: Central retinal vein occlusion (CRVO) leads to retinal ischemia, which then induces an upregulation of vascular endothelial growth factor (VEGF). The aim of this study was to determine whether a significant correlation exists between the ocular VEGF levels and the amplitudes and implicit times of different components of the electroretinogram (ERG) in eyes with a CRVO. METHODS: The medical records of the 20 consecutive patients who had macular edema secondary to CRVO and were examined at the Nagoya University Hospital from November 2008 to February 2010 were reviewed. Because all the patients were scheduled to receive an intravitreal injection of bevacizumab (IVB), it was possible to collect samples of the aqueous humor before the IVB. The correlation between the different components of the ERGs and the VEGF concentration in the aqueous was determined. RESULTS: The mean VEGF concentration of the aqueous humor was 416 pg/mL with a range of 100-1260 pg/mL. The b/a ratio of the single flash ERGs (P = 0.049; rho = -0.45), implicit times of the cone a-wave (P = 0.028; rho = 0.50), cone b-wave (P = 0.0059; rho = 0.63), and 30 Hz flicker ERGs (P = 0.0058; rho = 0.63) were significantly correlated with the VEGF concentration in the aqueous. CONCLUSIONS: The significant correlations between the different components of the ERGs and the aqueous VEGF concentration indicate that full-field ERGs can be used to detect the CRVO patients at a high risk of developing neovascularization of the iris. PMID- 21642627 TI - Ghrelin modulates physiologic and pathologic retinal angiogenesis through GHSR 1a. AB - PURPOSE: Vascular degeneration and the ensuing abnormal vascular proliferation are central to proliferative retinopathies. Given the metabolic discordance associated with these diseases, the authors explored the role of ghrelin and its growth hormone secretagogue receptor 1a (GHSR-1a) in proliferative retinopathy. METHODS: In a rat model of oxygen-induced retinopathy (OIR), the contribution of ghrelin and GHSR-1a was investigated using the stable ghrelin analogs [Dap3] ghrelin and GHRP6 and the GSHR-1a antagonists JMV-2959 and [D-Lys3]-GHRP-6. Plasma and retinal levels of ghrelin were analyzed by ELISA, whereas retinal expression and localization of GHSR-1a were examined by immunohistochemistry and Western blot analysis. The angiogenic and vasoprotective properties of ghrelin and its receptor were further confirmed in aortic explants and in models of vaso obliteration. RESULTS: Ghrelin is produced locally in the retina, whereas GHSR-1a is abundantly expressed in retinal endothelial cells. Ghrelin levels decrease during the vaso-obliterative phase and rise during the proliferative phase of OIR. Intravitreal delivery of [Dap3]-ghrelin during OIR significantly reduces retinal vessel loss when administered during the hyperoxic phase. Conversely, during the neovascular phase, ghrelin promotes pathologic angiogenesis through the activation of GHSR-1a. These angiogenic effects were confirmed ex vivo in aortic explants. CONCLUSIONS: New roles were disclosed for the ghrelin-GHSR-1a pathway in the preservation of retinal vasculature during the vaso-obliterative phase of OIR and during the angiogenic phase of OIR. These findings suggest that the ghrelin-GHSR-1a pathway can exert opposing effects on retinal vasculature, depending on the phase of retinopathy, and thus holds therapeutic potential for proliferative retinopathies. PMID- 21642629 TI - Relationship of stokes radius to the rate of diffusion across Bruch's membrane. AB - PURPOSE: To determine the effect of Stokes radius (R(S)) on the diffusion of molecules through Bruch's membrane (BM), and to establish a system suitable for the analysis of diffusion through small (<2 mm(2)) samples of BM. METHODS: Porcine BM/choroid (BM/Ch) was mounted in a modified Ussing chamber. A concentration gradient was simultaneously established for four tracers with R(S) values ranging from <1.0 to 6.15 nm. Samples were collected from both chambers at various time points up to 36 hours and the amount of each tracer was determined using quantitative gel exclusion chromatography. The integrity of samples was determined using scanning electron microscopy. RESULTS: BM/Ch mounted in the chamber exhibited no obvious damage even after 36 hours in the chamber. Flux was significantly (P < 0.05) greater in the BM to Ch direction than that in the Ch to BM direction for only two of the tracers: cytosine and RNase A. Flux also was dependent on R(S); cytosine, the smallest tracer (R(S) < 1 nm), exhibited the greatest flux and ferritin (R(S) = 6.15 nm) the least. Permeability coefficients for each tracer were determined and exhibited a power relationship with R(S). CONCLUSIONS: Flux was dependent on the direction of the concentration gradient and the R(S) of the individual tracers. We have successfully demonstrated that quantitative gel exclusion chromatography can be used to follow diffusion of a mixture of tracers across BM/Ch, and that we can measure flux across BM/Ch preparations with an exposed surface area as small as 1.8 mm(2). PMID- 21642628 TI - Exaggerated eye growth in IRBP-deficient mice in early development. AB - PURPOSE: Because interphotoreceptor retinoid-binding protein (IRBP) is expressed before being needed in its presumptive role in the visual cycle, we tested whether it controls eye growth during development. METHODS: The eyes of congenic IRBP knockout (KO) and C57BL/6J wild-type (WT) mice ranging in age from postnatal day (P)2 to P440 were compared by histology, laser micrometry, cycloplegic photorefractions, and partial coherence interferometry. RESULTS: The size and weight of IRBP KO mouse eyes were greater than those of the WT mouse, even before eye-opening. Excessive ocular enlargement started between P7 and P10, with KO retinal arc lengths becoming greater compared with WT from P10 through P30 (18%; P < 0.01). The outer nuclear layer (ONL) of KO retinas became 20% thinner between P12 to P25, and progressed to 38% thinner at P30. At P30, there were 30% fewer cones per vertical section in KO than in WT retinas. Bromodeoxyuridine (BrdU) labeling indicated the same number of retinal cells were born in KO and WT mice. A spike in apoptosis was observed in KO outer nuclear layer at P25. These changes in size were accompanied by a large decrease in hyperopic refractive error, which reached -4.56 +/- 0.70 diopters (D) versus +9.98 +/- 0.993 D (mean +/- SD) in WT, by postnatal day 60 (P60). CONCLUSIONS; In addition to its role in the visual cycle, IRBP is needed for normal eye development. How IRBP mediates ocular development is unknown. PMID- 21642630 TI - During glaucoma, alpha2-macroglobulin accumulates in aqueous humor and binds to nerve growth factor, neutralizing neuroprotection. AB - PURPOSE: Glaucoma is an optic neuropathy caused by the chronic and progressive death of retinal ganglion cells (RGCs), resulting in irreversible blindness. Ocular hypertension is a major risk factor, but RGC death often continues after ocular hypertension is normalized, and can take place with normal tension. Continuous RGC death was related in rodents and humans to the local upregulation of neurotoxic proteins, such as TNF-alpha. In rat models of glaucoma, ocular hypertension also upregulates the expression of alpha2-macroglobulin, which is neurotoxic. alpha2-macroglobulin upregulation in the retina is long-lived, even after high IOP is reduced with medication. alpha2-macroglobulin is examined as a possible biomarker in human glaucoma, and a possible neurotoxic mechanism of action is sought. METHODS: Quantitative Western blotting of alpha2-macroglobulin in samples obtained from aqueous humor (human and rat) and retina (rat) was conducted. Ex vivo neuronal survival assays and nerve growth factor-alpha2 macroglobulin binding studies using surface plasmon resonance were used. RESULTS: Increased soluble alpha2-macroglobulin protein is also present in the aqueous humor in a rat glaucoma model, as well as in the aqueous humor of human glaucoma patients but not in cataract patients. One mechanism by which alpha2 macroglobulin is neurotoxic is by inhibiting the neuroprotective activity of nerve growth factor via TrkA receptors. CONCLUSIONS: This work further documents a potential novel mechanism of RGC death and a potential biomarker or therapeutic target for glaucoma. PMID- 21642631 TI - Molecular analysis of Bardet-Biedl syndrome families: report of 21 novel mutations in 10 genes. AB - PURPOSE: Bardet-Biedl syndrome (BBS) is genetically heterogeneous with 15 BBS genes currently identified, accounting for approximately 70% of cases. The aim of our study was to define further the spectrum of BBS mutations in a cohort of 44 European-derived American, 8 Tunisian, 1 Arabic, and 2 Pakistani families (55 families in total) with BBS. METHODS: A total of 142 exons of the first 12 BBS causing genes were screened by dideoxy sequencing. Cases in which no mutations were found were then screened for BBS13, BBS14, BBS15, RPGRIP1L, CC2D2A, NPHP3, TMEM67, and INPP5E. RESULTS: Forty-three mutations, including 8 frameshift mutations, 10 nonsense mutations, 4 splice site mutations, 1 deletion, and 20 potentially or probably pathogenic missense variations, were identified in 46 of the 55 families studied (84%). Of these, 21 (2 frameshift mutations, 4 nonsense mutations, 4 splice site mutations, 1 deletion, and 10 missense variations) were novel. The molecular genetic findings raised the possibility of triallelic inheritance in 7 Caucasian families, 1 Arabian family, and 1 Tunisian patient. No mutations were detected for BBS4, BBS11, BBS13, BBS14, BBS15, RPGRIP1L, CC2D2A, NPHP3, TMEM67, or INPP5E. CONCLUSIONS: This mutational analysis extends the spectrum of known BBS mutations. Identification of 21 novel mutations highlights the genetic heterogeneity of this disorder. Differences in European and Tunisian patients, including the high frequency of the M390R mutation in Europeans, emphasize the population specificity of BBS mutations with potential diagnostic implications. The existence of some BBS cases without mutations in any currently identified BBS genes suggests further genetic heterogeneity. PMID- 21642632 TI - Safety evaluation of sunscreen formulations containing titanium dioxide and zinc oxide nanoparticles in UVB sunburned skin: an in vitro and in vivo study. AB - Sunscreens containing titanium dioxide (TiO(2)) and zinc oxide (ZnO) nanoparticles (NP) are effective barriers against ultraviolet B (UVB) damage to skin, although little is known about their disposition in UVB-damaged skin. Pigs were exposed to UVB that resulted in moderate sunburn. For in vitro studies, skin in flow-through diffusion cells were treated 24 h with four sunscreen formulations as follows: 10% coated TiO(2) in oil/water (o/w), 10% coated TiO(2) in water/oil (w/o), 5% coated ZnO in o/w, and 5% uncoated ZnO in o/w. TiO(2) (rutile, crystallite) primary particle size was 10 * 50 nm with mean agglomerates of 200 nm (range ca. 90 nm--460 nm); mean for ZnO was 140 nm (range ca. 60--200 nm). Skin was processed for light microscopy, scanning (SEM) and transmission electron microscopy (TEM), and time-of-flight secondary ion mass spectrometry (TOF-SIMS). UVB-exposed skin had typical sunburn histology. TEM showed TiO(2) NP 17 layers into stratum corneum (SC), whereas ZnO remained on the surface. TOF SIMS showed TiO(2) and ZnO epidermal penetration in both treatments. Perfusate analyzed by TEM/energy dispersive x-ray spectroscopy or inductively coupled plasma mass spectrometry detected no Ti or Zn, indicating minimal transdermal absorption. In vivo, skin was dosed at 24 h occluded with formulations and at 48 h. TiO(2) NP in o/w formulation penetrated 13 layers into UVB-damaged SC, whereas only 7 layers in normal skin; TiO(2) in w/o penetrated deeper in UVB-damaged SC. Coated and uncoated Zn NP in o/w were localized to the upper one to two SC layers in all skin. By SEM, NP were localized as agglomerates in formulation on the skin surface and base of hair. TOF-SIMS showed Ti within epidermis and superficial dermis, whereas Zn was limited to SC and upper epidermis in both treatments. In summary, UVB-damaged skin slightly enhanced TiO(2) NP or ZnO NP penetration in sunscreen formulations but no transdermal absorption was detected. PMID- 21642633 TI - Application of an integrated testing strategy to the U.S. EPA endocrine disruptor screening program. AB - New approaches to generating and evaluating toxicity data for chemicals are needed to cope with the ever-increasing demands of new programs. One such approach involves the use of an integrated testing and evaluation strategy based on the specific properties and activities of a chemical. Such an integrated strategy, whether applied to existing or future programs, can promote efficient use of resources and save animals. We demonstrate the utility of such a strategy by applying it to the current U.S. Environmental Protection Agency Endocrine Disruptor Screening Program (EDSP). Launched in October 2009, the EDSP utilizes a two-tiered approach, whereby each tier requires a battery of animal-intensive and expensive tests. Tier 1 consists of five in vitro and six in vivo assays that are intended to determine a chemical's potential to interact with the estrogen (E), androgen (A), or thyroid (T) hormone pathways. Tier 2 is proposed to consist of multigenerational reproductive and developmental toxicity tests in several species and is intended to determine whether a chemical can cause adverse effects resulting from E, A, or T modulation. In contrast to the existing EDSP structure, we show, using the pesticide atrazine as an example, that a multilevel testing framework combined with an integrated evaluation process would significantly increase efficiency by minimizing testing. PMID- 21642634 TI - Coevolution of telomeric repeats and telomeric repeat-specific non-LTR retrotransposons in insects. AB - In the telomeres of the silkworm Bombyx mori, telomeric repeat-specific non-long terminal repeat (LTR) retrotransposon SARTBm1 is accumulated in the TTAGG telomeric repeats. Here, we identify novel telomeric repeat-specific non-LTR retrotransposons, SARTTc family, from the red flour beetle Tribolium castaneum in the unconventional TCAGG telomeric repeats. To compare the sequence specificity of SARTBm1 and SARTTc1, we developed a comparable ex vivo retrotransposition assay. Both SARTBm1 and SARTTc1 preferred the telomeric sequence of their hosts, suggesting that the target specificity of these retrotransposons coevolved with their host's telomeric repeats. Swapping experiment indicated that the endonuclease domain is involved in recognizing the target sequence. Moreover, SARTBm1 proteins could retrotranspose 3'untranslated region (UTR) sequence of SARTTc1 as well as their own 3'UTR, whereas SARTTc1 proteins could only retrotranspose their own 3'UTRs. These results provide insights to the mechanism and divergence of sequence specificity and 3'UTR recognition in non-LTR retrotransposons. PMID- 21642635 TI - Effects of polybrominated diphenyl ethers on steroidogenesis in rat Leydig cells. AB - BACKGROUND: Polybrominated diphenyl ethers (PBDEs) are brominated flame retardants that have been defined as major environmental pollutants. While previous studies have found that PBDEs may enhance the levels of sex-steroid hormones, their effects on testosterone secretion from rat Leydig cells are unclear. This study investigated the effects and mechanisms of PBDE-710, a mixture of tetra- and penta-PBDEs, on testosterone biosynthesis in rat Leydig cells. METHODS: Leydig cells from adult male rats were challenged with different concentrations of PBDE-710 (0.5-15 ng/ml) to evaluate the effects on testosterone steroidogenesis. Concentrations of testosterone and of cAMP and pregnenolone in medium were measured by radioimmunoassay (RIA) and by enzyme-linked immunosorbent assay, respectively. Nuclear translocation of protein kinase A alpha (PKAalpha) was determined by immunofluorence assay and western blot assay, and the mRNA expression of steroidogenic acute regulatory protein (StAR) was analyzed by quantitative real-time polymerase chain reaction. RESULTS: In this in vitro study, PBDE-710 (5 or 15 ng/ml) increased basal testosterone secretion and cAMP production by 3- and 2-fold, respectively. The stimulatory effect was abolished by adenylyl cyclase inhibitor. Enzyme activity of CYP11A1, as determined by the pregnenolone concentration, was stimulated by PBDE-710 treatment. Furthermore, nuclear translocation of PKAalpha was increased by 20% and StAR gene expression was elevated by 4-fold after PBDE-710 treatment. CONCLUSIONS: These results suggest that low concentrations of PBDE-710 could stimulate testosterone secretion by acting directly on Leydig cells to activate the cAMP pathway and increase expression of StAR. PMID- 21642636 TI - Ovarian cancer-associated polymorphisms in the BNC2 gene among women with endometriosis. AB - BACKGROUND: Endometriosis is a common benign gynaecological disease. Epidemiological studies have demonstrated associations between endometriosis and ovarian cancer. Recent genome-wide association studies of ovarian cancer have identified several single nucleotide polymorphisms (SNPs) in the Basonuclin 2 (BNC2) gene. In this study, we investigated these polymorphism in women with endometriosis. METHODS: Six SNPs in and upstream of the BNC2 gene (rs3814113, rs4445329, rs10962656, rs12379183, rs10756819 and rs1339552) were investigated using TaqMan allelic discrimination analysis in a Caucasian population (cases: 798, controls: 351). Allelic frequencies were used as main outcome measure. RESULTS: No associations were observed between the analysed SNPs and endometriosis. CONCLUSIONS: Our results suggest that the analysed polymorphisms in the BNC2 gene are unlikely to contribute to the previously reported risk of ovarian cancer in women with endometriosis. PMID- 21642637 TI - Uterine artery embolization for symptomatic fibroids: long-term changes in disease-specific symptoms and quality of life. AB - BACKGROUND: To investigate the long-term changes in health-related quality of life (HRQOL) after uterine artery embolization (UAE) for symptomatic fibroids, we conducted a prospective cohort study. METHODS: Eighty-two women completed the validated uterine fibroid symptom and QOL (UFS-QOL) questionnaire before UAE. Short-term results after a median of 8 months (range: 3-20) have been published previously. Patients were asked to complete the questionnaire again after a median of 6.3 years (range: 5-7.6). Secondary outcome measures were the frequency of additional surgical or endovascular procedures due to treatment failure and the menstrual status. RESULTS: A total of 4/82 patients (5%) were lost to follow up. Of the remaining 78 patients, 11 underwent surgery or repeat UAE (hysterectomy n = 6, myomectomy n = 1, UAE n = 4) at a median of 13 months (range: 5-70) after UAE. Two patients failed clinically but did not undergo a second intervention. The overall treatment failure rate 6 years after UAE was 17%. Clinical long-term follow-up regarding symptom control and quality of life was available in the remaining 65 patients. Symptom severity scores decreased from a median of 37.50 (quartile range, QR: 28.13-53.13) to 0.00 (QR: 0.00-10.94) (P < 0.001), whereas the HRQOL total score increased from a median of 64.66 (QR: 46.34-79.10) to 100.00 (QR: 96.12-100.00) (P < 0.001). Both scores also improved significantly compared with short-term results (P = 0.006 and P = 0.041). Permanent amenorrhea was observed in five patients at a median of 18 months (range: 10-46) after UAE and at a mean patient age of 50 years. CONCLUSIONS: UAE leads to durable relief of fibroid-related symptoms and sustained improvement in HRQOL. After 6 years, clinical failure can be expected in 17%, and most of these patients require secondary invasive treatment. PMID- 21642638 TI - Oral contraceptives and endometriosis: the past use of oral contraceptives for treating severe primary dysmenorrhea is associated with endometriosis, especially deep infiltrating endometriosis. AB - BACKGROUND: The relationship between the use of oral contraception (OC) and endometriosis remains controversial. We therefore compared various characteristics of OC use and the surgical diagnosis of endometriosis histologically graded as superficial peritoneal endometriosis (SUP), ovarian endometrioma (OMA) or deep infiltrating endometriosis (DIE). METHODS: This cross sectional study included 566 patients without visible endometriosis at surgery as controls, and 410 patients with histologically proven endometriosis, categorized by their worst lesions as SUP n = 47, OMA n = 120 and DIE n = 243. Personal data, including on OC use, were prospectively collected during standardized interviews. Statistical analysis was performed using unconditional logistic regression. RESULTS: Past OC users had an increased incidence of endometriosis (adjusted odd ratios (OR) = 2.79, 95% confidence interval (CI) 1.74-5.12, P = 0.002) of any revised American Fertility Society stage. Women who had previously used OC for severe primary dysmenorrhea were even more frequently diagnosed with endometriosis (adjusted OR = 5.6, 95% CI 3.2-9.8), especially for DIE (adjusted OR = 16.2, 95% CI 7.8-35.3). Women who had previously used OC for other reasons also had an increased risk of endometriosis, but to a lesser extent (adjusted OR = 2.6, 95% CI 1.8-4.1). The age at which OC was initiated, duration of OC use and free interval from last OC use were not significantly different between control and endometriosis women, irrespective of histological grading. Current OC users did not show an increased prevalence of endometriosis (OR = 1.22, 95% CI 0.6 2.52). CONCLUSIONS: Our data indicate that a history of OC use for severe primary dysmenorrhea is associated with surgical diagnosis of endometriosis, especially DIE, later in life. However, this does not necessarily mean that use of OC increases the risk of developing endometriosis. Past use of OC for primary dysmenorrhea may serve as a marker for women with endometriosis and DIE. PMID- 21642639 TI - Tubal factor infertility is associated with antibodies against Chlamydia trachomatis heat shock protein 60 (HSP60) but not human HSP60. AB - BACKGROUND: Serum antibodies against major outer membrane protein (MOMP) and heat shock protein 60 (HSP60) from Chlamydia trachomatis are correlated with sequelae following infection. Since bacterial and human HSP60 share considerable sequence homology, cross-reactivity to human HSP60 is suggested as being involved in tubal factor infertility (TFI). The aim was to investigate whether antibodies to human HSP60 are associated with TFI, and to evaluate antibody testing in TFI diagnosis. METHODS: Serum levels of antibodies against chlamydial MOMP and HSP60 from C. trachomatis, Salmonella enterica Enteritidis, Campylobacter jejuni and human HSP60 were analysed by enzyme-linked immunosorbent assay in three groups of infertile women: women with TFI (n = 70), controls with normal fallopian tubes (control group 1, n = 92) and a subgroup of women with normal fallopian tubes and sero-positive for either chlamydial MOMP or chlamydial HSP60 (control group 2, n = 28). RESULTS: Serum levels of immunoglobulin (Ig)G1 and IgG3 antibodies against MOMP and HSP60 from C. trachomatis were elevated in patients with TFI compared with non-TFI individuals (group 1; P < 0.001), while levels of IgG3 against MOMP and IgG1 against HSP60 were higher in the TFI group compared with control group 2 (P = 0.04 and P = 0.03, respectively). Levels of antibodies against human HSP60 did not differ between groups. CONCLUSIONS: Our findings confirm an association between TFI and antibodies to MOMP and HSP60 from C. trachomatis, suggesting antibody testing as a supplement in TFI diagnosis. No connection was observed between TFI and antibodies to human HSP60, pointing to an infectious rather than an autoimmune inflammation as the cause of TFI. PMID- 21642640 TI - Mass spectrometry-based proteomics using Q Exactive, a high-performance benchtop quadrupole Orbitrap mass spectrometer. AB - Mass spectrometry-based proteomics has greatly benefitted from enormous advances in high resolution instrumentation in recent years. In particular, the combination of a linear ion trap with the Orbitrap analyzer has proven to be a popular instrument configuration. Complementing this hybrid trap-trap instrument, as well as the standalone Orbitrap analyzer termed Exactive, we here present coupling of a quadrupole mass filter to an Orbitrap analyzer. This "Q Exactive" instrument features high ion currents because of an S-lens, and fast high-energy collision-induced dissociation peptide fragmentation because of parallel filling and detection modes. The image current from the detector is processed by an "enhanced Fourier Transformation" algorithm, doubling mass spectrometric resolution. Together with almost instantaneous isolation and fragmentation, the instrument achieves overall cycle times of 1 s for a top 10 higher energy collisional dissociation method. More than 2500 proteins can be identified in standard 90-min gradients of tryptic digests of mammalian cell lysate- a significant improvement over previous Orbitrap mass spectrometers. Furthermore, the quadrupole Orbitrap analyzer combination enables multiplexed operation at the MS and tandem MS levels. This is demonstrated in a multiplexed single ion monitoring mode, in which the quadrupole rapidly switches among different narrow mass ranges that are analyzed in a single composite MS spectrum. Similarly, the quadrupole allows fragmentation of different precursor masses in rapid succession, followed by joint analysis of the higher energy collisional dissociation fragment ions in the Orbitrap analyzer. High performance in a robust benchtop format together with the ability to perform complex multiplexed scan modes make the Q Exactive an exciting new instrument for the proteomics and general analytical communities. PMID- 21642641 TI - Enforced relocation of older people when Care Homes close: a question of life and death? AB - Care Homes are usually seen as the last refuge for older people but residents are sometimes required to move between homes for administrative purposes. There is concern that such moves threaten their well-being and survival. Relocations have been contested repeatedly in court. A recent ruling and its review of case-law and literature provides guidance for practitioners who may be consulted for advice in this demanding situation. PMID- 21642642 TI - Ultrasound assessment of cranial spread during caudal blockade in children: the effect of different volumes of local anaesthetics. AB - BACKGROUND: Despite the large amount of literature on caudal anaesthesia in children, the issue of volume of local anaesthetics and cranial spread is still not settled. Thus, the aim of the present prospective randomized study was to evaluate the cranial spread of caudally administered local anaesthetics in children by means of real-time ultrasound, with a special focus on the effects of using different volumes of local anaesthetics. METHODS: Seventy-five children, 1 month to 6 yr, undergoing inguinal hernia repair or more distal surgery were randomized to receive a caudal block with 0.7, 1.0, or 1.3 ml kg(-1) ropivacaine. The cranial spread of the local anaesthetic within the spinal canal was assessed by real-time ultrasound scanning; the absolute cranial segmental level and the cranial level relative to the conus medullaris were determined. RESULTS: All the blocks were judged to be clinically successful. A significant correlation was found between the injected volume and the cranial level reached by the local anaesthetic both with regards to the absolute cranial segmental level and the cranial level relative to the conus medullaris. CONCLUSIONS: The main finding of the present study was positive, but numerically small correlation between injected volumes of local anaesthetic and the cranial spread of caudally administered local anaesthetics. Therefore, the prediction of the cranial spread of local anaesthetic, depending on the injected volume of the local anaesthetic, was not possible. EudraCT Number: 2008-007627-40. PMID- 21642643 TI - Cardiac output can be measured with the transpulmonary thermodilution method in a paediatric animal model with a left-to-right shunt. AB - BACKGROUND: The transpulmonary thermodilution (TPTD) technique for measuring cardiac output (CO) has never been validated in the presence of a left-to-right shunt. METHODS: In this experimental, paediatric animal model, nine lambs with a surgically constructed aorta-pulmonary left-to-right shunt were studied under various haemodynamic conditions. CO was measured with closed and open shunt using the TPTD technique (CO(TPTD)) with central venous injections of ice-cold saline. An ultrasound transit time perivascular flow probe around the main pulmonary artery served as the standard reference measurement (CO(MPA)). RESULTS: Seven lambs were eligible for further analysis. Mean (sd) weight was 6.6 (1.6) kg. The mean CO(MPA) was 1.21 litre min(-1) (range 0.61-2.06 l min(-1)) with closed shunt and 0.93 litre min(-1) (range 0.48-1.45 litre min(-1)) with open shunt. The open shunt resulted in a mean Q(p)/Q(s) ratio of 1.8 (range 1.6-2.4). The bias between the two CO methods was 0.17 litre min(-1) [limits of agreement (LOA) of 0.27 litre min(-1)] with closed shunt and 0.14 litre min(-1) (LOA of 0.32 litre min( 1)) with open shunt. The percentage errors were 22% with closed shunt and 34% with open shunt. The correlation (r) between the two methods was 0.93 (P<0.001) with closed shunt and 0.86 (P<0.001) with open shunt. The correlation (r) between the two methods in tracking changes in CO (DeltaCO) during the whole experiment was 0.94 (P<0.0001). CONCLUSIONS: The TPTD technique is a feasible method of measuring CO in paediatric animals with a left-to-right shunt. PMID- 21642644 TI - Effect of phenylephrine and ephedrine bolus treatment on cerebral oxygenation in anaesthetized patients. AB - BACKGROUND: How phenylephrine and ephedrine treatments affect global and regional haemodynamics is of major clinical relevance. Cerebral tissue oxygen saturation (Sct(O2) )-guided management may improve postoperative outcome. The physiological variables responsible for Sct(O2) changes induced by phenylephrine and ephedrine bolus treatment in anaesthetized patients need to be defined. METHODS: A randomized two-treatment cross-over trial was conducted: one bolus dose of phenylephrine (100-200 ug) and one bolus dose of ephedrine (5-20 mg) were given to 29 ASA I-III patients anaesthetized with propofol and remifentanil. , mean arterial pressure (MAP), cardiac output (CO), and other physiological variables were recorded before and after treatments. The associations of changes were analysed using linear-mixed models. RESULTS: The CO decreased significantly after phenylephrine treatment [?CO = -2.1 (1.4) litre min(-1), P<0.001], but was preserved after ephedrine treatment [?CO = 0.5 (1.4) litre min(-1), P>0.05]. The was significantly decreased after phenylephrine treatment [? = -3.2 (3.0)%, P<0.01] but preserved after ephedrine treatment [? = 0.04 (1.9)%, P>0.05]. CO was identified to have the most significant association with (P<0.001). After taking CO into consideration, the other physiological variables, including MAP, were not significantly associated with (P>0.05). CONCLUSIONS: Associated with changes in CO, decreased after phenylephrine treatment, but remained unchanged after ephedrine treatment. The significant correlation between CO and implies a cause effect relationship between global and regional haemodynamics. PMID- 21642645 TI - Strategies for locating the female gamete: the importance of measuring sperm trajectories in three spatial dimensions. AB - The spermatozoon must find its female gamete partner and deliver its genetic material to generate a new individual. This requires that the spermatozoon be motile and endowed with sophisticated swimming strategies to locate the oocyte. A common strategy is chemotaxis, in which spermatozoa detect and follow a gradient of chemical signals released by the egg and its associated structures. Decoding the female gamete's positional information is a process that spermatozoa undergo in a three-dimensional (3D) space; however, due to their speed and small size, this process has been studied almost exclusively in spermatozoa restricted to swimming in two dimensions (2D). This review examines the relationship between the mechanics of sperm propulsion and the physiological function of these cells in 3D. It also considers whether it is possible to derive all the 3D sperm swimming characteristics by extrapolating from 2D measurements. It is concluded that full insight into flagellar beat dynamics, swimming paths and chemotaxis under physiological conditions will eventually require quantitative imaging of flagellar form, ion flux changes, cell trajectories and modelling of free swimming spermatozoa in 3D. PMID- 21642647 TI - Measurements of 222Rn activity concentration in domestic water sources in Penang, northern peninsular Malaysia. AB - Measurements of (222)Rn activity concentration were carried out in 39 samples collected from the domestic and drinking water sources used in the island and mainland of Penang, northern peninsular, Malaysia. The measured activity concentrations ranged from 7.49 to 26.25 Bq l(-1), 0.49 to 9.72 Bq l(-1) and 0.58 to 2.54 Bq l(-1) in the raw, treated and bottled water samples collected, respectively. This indicated relatively high radon concentrations compared with that from other parts of the world, which still falls below the WHO recommended treatment level of 100 Bq l(-1). From this data, the age-dependent associated committed effective doses due to the ingestion of (222)Rn as a consequence of direct consumption of drinking water were calculated. The committed effective doses from (222)Rn resulting from 1 y's consumption of these water were estimated to range from 0.003 to 0.048, 0.001 to 0.018 and 0.002 to 0.023 mSv y(-1), for age groups 0-1, 2-16 and >16 y, respectively. PMID- 21642646 TI - Rediscovering sperm ion channels with the patch-clamp technique. AB - Upon ejaculation, mammalian spermatozoa have to undergo a sequence of physiological transformations within the female reproductive tract that will allow them to reach and fertilize the egg. These include initiation of motility, hyperactivation of motility and perhaps chemotaxis toward the egg, and culminate in the acrosome reaction that permits sperm to penetrate the protective vestments of the egg. These physiological responses are triggered through the activation of sperm ion channels that cause elevations of sperm intracellular pH and Ca(2+) in response to certain cues within the female reproductive tract. Despite their key role in sperm physiology and their absolute requirement for the process of fertilization, sperm ion channels remain poorly understood due to the extreme difficulty in application of the patch-clamp technique to spermatozoa. This review covers the topic of sperm ion channels in the following order: first, we discuss how the intracellular Ca(2+) and pH signaling mediated by sperm ion channels controls sperm behavior during the process of fertilization. Then, we briefly cover the history of the methodology to study sperm ion channels, which culminated in the recent development of a reproducible whole-cell patch-clamp technique for mouse and human cells. We further discuss the main approaches used to patch-clamp mature mouse and human spermatozoa. Finally, we focus on the newly discovered sperm ion channels CatSper, KSper (Slo3) and HSper (H(v)1), identified by the sperm patch-clamp technique. We conclude that the patch-clamp technique has markedly improved and shifted our understanding of the sperm ion channels, in addition to revealing significant species-specific differences in these channels. This method is critical for identification of the molecular mechanisms that control sperm behavior within the female reproductive tract and make fertilization possible. PMID- 21642648 TI - Acupuncture sensation during ultrasound guided acupuncture needling. AB - BACKGROUND: Although acupuncture sensation (also known as de qi) is a cornerstone of traditional acupuncture therapy, most research has accepted the traditional method of defining acupuncture sensation only through subjective patient reports rather than on any quantifiable physiological basis. PURPOSE: To preliminarily investigate the frequency of key sensations experienced while needling to specific, quantifiable tissue levels (TLs) guided by ultrasound (US) imaging. METHODS: Five participants received needling at two acupuncture points and two control points at four TLs. US scans were used to determine when each TL was reached. Each volunteer completed 32 sets of modified Southampton Needle Sensation Questionnaires. Part one of the study tested sensations experienced at each TL and part two compared the effect of oscillation alone versus oscillation+rotation. RESULTS: In all volunteers, the frequency of pricking, sharp sensations was significantly greater in shallower TLs than deeper (p=0.007); the frequency of sensations described as deep, dull and heavy, as spreading, and as electric shocks was significantly greater in deeper TLs than shallower (p=0.002). Sensations experienced did not significantly differ between real and control points within each of three TLs (p>0.05) except TL 4 (p=0.006). The introduction of needle rotation significantly increased deep, dull, heavy sensations, but not pricking and sharp sensations; within each level, the spectrum of sensation experienced during both oscillation+rotation and oscillation alone did not significantly differ between acupuncture and control points. CONCLUSION: The preliminary study indicates a strong connection between acupuncture sensation and both tissue depth and needle rotation. Furthermore, the new methodology has been proven feasible. A further study with an objective measurement is warranted. PMID- 21642649 TI - Pharmacokinetics of the raltegravir/maraviroc/etravirine combination. PMID- 21642650 TI - A quality improvement programme to increase compliance with an anti-infective prescribing policy. AB - OBJECTIVES: The UK Department of Health has made recommendations on safe and appropriate prescribing of anti-infectives. In response, we reviewed our anti infective policies to ensure they were in line with best practice. As a result, a new adult anti-infective policy was launched. To help facilitate its implementation, a quality improvement programme was established, with the aim of achieving >90% compliance with the new policy. METHODS: Patients under the care of the medical admissions teams who had been prescribed one or more systemic anti infectives between January and November 2008 were included in the study. Study pharmacists collected data daily on all patients, including the anti-infective(s) prescribed and indication(s) documented on either the patient's drug prescription chart or health records. A definition of compliance was developed, which required documented indication(s) and associated anti-infectives to match the anti infective policy. A baseline compliance level was established; we then implemented a series of interventions using the plan-do-study-act ('PDSA') approach to monitor and improve compliance. Three overlapping intervention phases were retrospectively identified: raising awareness; education; and weekly feedback of results in the form of run charts distributed to medical teams. RESULTS: Over the 11 month study period, compliance with the policy increased from 30% to 71%. Since 2008, we have seen the average compliance increase year-on year to over 90% in 2010 using a sustainable once weekly data collection model. CONCLUSIONS: This study shows that it is possible to use quality improvement methodology to support antimicrobial stewardship within existing resources and suggests that an improvement in policy compliance can be both achieved and sustained. PMID- 21642651 TI - The multiresistant Acinetobacter baumannii European clone I type strain RUH875 (A297) carries a genomic antibiotic resistance island AbaR21, plasmid pRAY and a cluster containing ISAba1-sul2-CR2-strB-strA. PMID- 21642652 TI - Analysis of risk factors for malignant Mediterranean spotted fever indicates that fluoroquinolone treatment has a deleterious effect. AB - OBJECTIVES: To identify risk factors for malignant Mediterranean spotted fever (MSF) caused by Rickettsia conorii conorii. PATIENTS AND METHODS: Epidemiological, clinical and biological characteristics as well as risk factors (including treatment regimens) for severe MSF cases were analysed retrospectively. A patient with two or more organ dysfunctions or patient death was defined as a severe case. RESULTS: During the study period (January 1999 to December 2009), 161 MSF cases were referred to our centre for rickettsioses. Twenty-six cases (16.1%) were considered severe, which is 3-fold higher than in our previous studies. The clinical and laboratory findings were comparable to those reported elsewhere except that the type of antibiotic treatment was associated with disease severity. Doxycycline administration prior to deterioration of disease (in 31 patients) protected patients from development of severe MSF [relative risk (RR) 0.248, 95% confidence interval (CI) 0.08-0.76] and induced earlier defervescence compared with the other treatment regimens (3.02 +/ 2.2 days versus 7.1 +/- 6.57 days, P = 0.021). In contrast, fluoroquinolone treatment (in 21 patients) was significantly and independently associated with MSF severity (RR 2.53, 95% CI 1.40-4.55) and was associated with a significantly longer hospital stay. CONCLUSIONS: In this retrospective study fluoroquinolone treatment was associated with increased MSF disease severity. Fluoroquinolones have been previously associated with treatment failure in typhus and scrub typhus cases. Thus, we do not recommend the use of fluoroquinolones to treat rickettsial diseases. PMID- 21642653 TI - Bilateral endogenous endophthalmitis secondary to Candida albicans. PMID- 21642654 TI - Acute respiratory distress syndrome and acute lung injury. AB - Acute respiratory distress syndrome (ARDS) is a life threatening respiratory failure due to lung injury from a variety of precipitants. Pathologically ARDS is characterised by diffuse alveolar damage, alveolar capillary leakage, and protein rich pulmonary oedema leading to the clinical manifestation of poor lung compliance, severe hypoxaemia, and bilateral infiltrates on chest radiograph. Several aetiological factors associated with the development of ARDS are identified with sepsis, pneumonia, and trauma with multiple transfusions accounting for most cases. Despite the absence of a robust diagnostic definition, extensive epidemiological investigations suggest ARDS remains a significant health burden with substantial morbidity and mortality. Improvements in outcome following ARDS over the past decade are in part due to improved strategies of mechanical ventilation and advanced support of other failing organs. Optimal treatment involves judicious fluid management, protective lung ventilation with low tidal volumes and moderate positive end expiratory pressure, multi-organ support, and treatment where possible of the underlying cause. Moreover, advances in general supportive measures such as appropriate antimicrobial therapy, early enteral nutrition, prophylaxis against venous thromboembolism and gastrointestinal ulceration are likely contributory reasons for the improved outcomes. Although therapies such as corticosteroids, nitric oxide, prostacyclins, exogenous surfactants, ketoconazole and antioxidants have shown promising clinical effects in animal models, these have failed to translate positively in human studies. Most recently, clinical trials with beta2 agonists aiding alveolar fluid clearance and immunonutrition with omega-3 fatty acids have also provided disappointing results. Despite these negative studies, mortality seems to be in decline due to advances in overall patient care. Future directions of research are likely to concentrate on identifying potential biomarkers or genetic markers to facilitate diagnosis, with phenotyping of patients to predict outcome and treatment response. Pharmacotherapies remain experimental and recent advances in the modulation of inflammation and novel cellular based therapies, such as mesenchymal stem cells, may reduce lung injury and facilitate repair. PMID- 21642655 TI - Re: The use and impact of national confidential enquiries in high-income countries BMJ Qual Saf 2011;20:38-45. PMID- 21642657 TI - Claudin and p53 expression in vulvar lichen sclerosus and squamous-cell carcinoma. AB - AIMS: Vulvar squamous-cell carcinoma (SCC) is a rare gynaecological cancer. Vulvar SCC has been shown to develop from vulvar intraepithelial neoplasias, which are related to lichen sclerosus (LS). Most studies to date have compared vulvar SCC with LS only morphologically, but no detailed molecular analysis has been performed. The objective was to compare claudin and p53 expression in these diseases and determine if there was any association with expression and vulvar SCC progression. METHODS: Immunohistochemical analysis was performed in order to determine expression of p53 and claudin 1, 2, 3, 4, 5, 7 and 11 in human vulvar tissue samples from LS, SCC and control patients. RESULTS: Claudin 1, 2, 3, 4 and 5 were expressed comparably in the three groups. Claudin 7 and 11 expression was significantly decreased in LS and SCC samples compared with the control group. Expression of p53 was significantly increased in SCC and LS patient samples compared with the control group. CONCLUSIONS: Claudin 7 and 11 were not expressed in LS and SCC. However, there was no significant difference in expression of any of the claudins between the LS and SCC samples. Furthermore, p53 expression is the highest in SCC patients and lowest in the control group. However, expression of p53 did not vary between samples from isolated LS and LS associated SCC patients, suggesting that increased p53 expression is not the determining factor in the progression of LS lesions to SCC. PMID- 21642658 TI - Prognostic molecular markers in women aged 35 years or younger with breast cancer: is there a difference from the older patients? AB - BACKGROUND: Women aged <= 35 years with breast cancer have a poor prognosis, but their prognostic factors have not been clearly defined. AIMS: To evaluate whether the molecular markers used in age-unspecified breast cancer could also be applied to women <= 35 years. METHODS: Archival tumours from patients aged <= 35 years with stage I-III breast cancer were collected. Oestrogen receptor (ER), progesterone receptor (PR), HER2, Ki67 and P53 protein expression profiles in paraffin-embedded tissue sections were determined by immunohistochemistry. Tumours with an HER2 score of 2+ were further evaluated by fluorescence in situ hybridisation. Mutational analysis of exons 4-9 of the TP53 gene and exons 9 and 20 of the PIK3CA gene was carried out using direct sequencing analysis. RESULTS: 116 patients with a median follow-up duration of 62.7 months were included. In addition to tumour size and axillary lymph node status, univariate analysis showed that high Ki67 expression, ER-negative, HER2 overexpression, and TP53 mutations were associated with shorter overall survival. Multivariate analysis showed that high Ki67 expression (HR=3.93, p=0.005), HER2 overexpression (HR=3.21, p=0.013) and TP53 mutations (HR=4.44, p=0.005) were associated with shorter overall survival. PR expression and PIK3CA mutations were not associated with survival. CONCLUSIONS: For women <= 35 years, TP53 mutations, Ki67 and HER2 expressions are strong prognostic factors. The limited prognostic value of hormone receptors suggests that the prognostic markers used in age-unspecified breast cancer may not be completely fit for this population. PMID- 21642659 TI - Syringoid eccrine carcinoma: a clinicopathological and immunohistochemical study of four cases. AB - BACKGROUND: Syringoid eccrine carcinoma (SEC) is a rare malignant adnexal tumour with variable presentations. AIM: To examine the clinicopathological and immunohistochemical features of SEC. METHODS: Four cases were reviewed by three dermatopathologists and the immunohistochemical profile was examined using antibodies against CK5/6, CK7, CK14, CK20, LMWK, HMWK, EMA, mCEA, p63, ER, PR, AR, S-100 and Ber-EP4. RESULTS: The cases occurred in two men and two women, ranging in age from 61 to 87 years (mean 68.5). Two of the lesions were from the face and two from the trunk. All four lesions were composed of an atypical infiltrative mass with syringoma-like tadpole morphology with ductular differentiation and prominent desmoplasia. Three cases demonstrated perineural invasion and two had positive lymph node metastases. Immunostaining was variable. Immunohistochemistry positivity was as follows: three out of four cases were positive for CK5/6, CK7 (2/4), CK14 (1/3), CK20 (0/2), HMWK (0/2), LMWK (1/2), EMA (3/4), mCEA (4/4), p63 (2/3), ER (2/3), PR (1/2), AR (0/3), S-100 (0/3) and Ber-EP4 (2/2). CONCLUSION: SEC can present on the trunk and are not limited to the head and neck region. In addition to syringoma-like tadpole structures and glandular differentiation, these tumours can also exhibit squamoid and cribriform growth patterns. Immunostaining in SEC is variable and this variability is believed to stem from this tumour's ability to differentiate along multiple routes, including sweat secretory and/or ductal differentiation. PMID- 21642660 TI - Mechanisms and management of the heart in myotonic dystrophy. PMID- 21642661 TI - Cardiac masses: an integrative approach using echocardiography and other imaging modalities. PMID- 21642662 TI - Clopidogrel loading dose in patients undergoing elective percutaneous coronary intervention. PMID- 21642663 TI - Neuroimaging signatures and cognitive correlates of the montreal cognitive assessment screen in a nonclinical elderly sample. AB - The Montreal Cognitive Assessment (MoCA) screen was developed as a brief instrument to identify mild cognitive impairment and dementia among older individuals. To date, limited information is available regarding the neuroimaging signatures associated with performance on the scale, or the relationship between the MoCA and more comprehensive cognitive screening measures. The present study examined performances on the MoCA among 111 non-clinical older adults (ages 51 85) enrolled in a prospective study of cognitive aging. Participants were administered the MoCA, Mini-Mental State Exam (MMSE), and the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS). A subset of participants (N = 69) underwent structural 3 T magnetic resonance imaging (MRI) to define the volumes of total frontal gray matter, total hippocampus, T2-weighted subcortical hyperintensities (SH), and total brain volume. The results revealed significant correlations between the total score on the MoCA and total score on the RBANS and MMSE, though the strength of the correlations was more robust between the MoCA and the RBANS. Modest correlations between individual subscales of the MoCA and neuroimaging variables were evident, but no patterns of shared variance emerged between the MoCA total score and neuroimaging indices. In contrast, total brain volume correlated significantly with total score on the RBANS. These results suggest that additional studies are needed to define the significance of MoCA scores relative to brain integrity among an older population. PMID- 21642664 TI - Base rates of post-concussive symptoms in a nonconcussed multicultural sample. AB - The purpose of this preliminary study was to investigate differences in base rates of post-concussion syndrome (PCS) symptomatology in healthy individuals separated by cultural identity and language. In this study, we sought to determine if differences exist in terms of base-rate endorsement of PCS symptomatology in healthy individuals and whether culture and language-based specific clusters of PCS symptomatology exist in healthy individuals. A total of 151 participants, consisting of 33 Caucasians, 49 Chinese, Filipinos, and Southeast Asians, 43 Arabs, West Asians, and South Asians, and 26 participants of African descent completed four questionnaires and two subtests of a test of verbal ability. We found that the occurrence of PCS symptoms did not differ by culture and language-based groups in general, but that there were differences between groups in the base rates of individual symptom endorsement. Our preliminary findings illustrate that cultural and linguistic background may play a moderating role in the endorsement of PCS symptomatology and that false positive conclusions may be reached disproportionally between groups. Our results underscore the importance of considering the influence of culture and language, in conjunction with a growing list of factors that can influence, maintain, and/or mimic persistent PCS. PMID- 21642665 TI - Combined immunotherapy with low-dose IL-2 plus IFN-alpha for metastatic renal cell carcinoma: survival benefit for selected patients with lung metastasis and serum sodium level. AB - OBJECTIVE: To clarify the survival benefit of immunotherapy for renal cell carcinoma patients with lung metastasis using low-dose interleukin-2 plus interferon-alpha, we examined survival outcomes and factors associated with prognosis. METHODS: This was a multicenter prospective study. Nephrectomized renal cell carcinoma patients with lung metastasis were treated with interleukin 2 (0.7 * 10(6) unit, 5 days a week) and interferon-alpha (6 * 10(6) IU, 3 days a week) for the first 8 weeks, and then with both interleukin-2 and interferon alpha, 2 or 3 days a week for 16 additional weeks. RESULTS: Median follow-up period for 42 patients was 28.3 months (range: 4.2-43.8). Two-year overall survival rate was 82% and the probability of 3 year survival rate was 71%. Median progression-free survival was 10.4 months. While no difference was found in survival among patients assessed as complete response, partial response and no change, survival of patients assessed as NC or better was significantly better than those assessed as progressive disease (P < 0.0001). Furthermore, multivariate analyses identified pre-treatment serum sodium (P = 0.004) as an independent prognostic factor. The sodium level was also statistically associated with tumor response (p = 0.035). Patients with normal sodium level survived significantly longer (P = 0.0005) than those with low sodium level showing median survival of 12.2 months. CONCLUSIONS: Combination immunotherapy with low-dose interleukin-2 plus interferon-alpha showed survival benefit for patients with lung metastasis whose tumor responded as no change or better. This combination immunotherapy could be beneficial for patients selected by metastatic organ and their pre-treatment serum sodium level. PMID- 21642666 TI - The cover: Penumbra. PMID- 21642667 TI - A piece of my mind: Way back when. PMID- 21642668 TI - Debate continues on use of PSA testing for early detection of prostate cancer. PMID- 21642669 TI - Retinoblastoma therapy delivers power of chemotherapy with surgical precision. PMID- 21642670 TI - Studies probe role of telomere length in predicting, modulating cancer risk. PMID- 21642671 TI - Axillary vs sentinel lymph node dissection for invasive breast cancer. PMID- 21642672 TI - Axillary vs sentinel lymph node dissection for invasive breast cancer. PMID- 21642673 TI - Axillary vs sentinel lymph node dissection for invasive breast cancer. PMID- 21642674 TI - Axillary vs sentinel lymph node dissection for invasive breast cancer. PMID- 21642675 TI - Axillary vs sentinel lymph node dissection for invasive breast cancer. PMID- 21642676 TI - Axillary vs sentinel lymph node dissection for invasive breast cancer. PMID- 21642677 TI - Bevacizumab and cancer treatment-related mortality. PMID- 21642678 TI - Bevacizumab and cancer treatment-related mortality. PMID- 21642679 TI - Bevacizumab and cancer treatment-related mortality. PMID- 21642680 TI - Antibiotic resistance and the future. PMID- 21642682 TI - Cancer risks associated with germline mutations in MLH1, MSH2, and MSH6 genes in Lynch syndrome. AB - CONTEXT: Providing accurate estimates of cancer risks is a major challenge in the clinical management of Lynch syndrome. OBJECTIVE: To estimate the age-specific cumulative risks of developing various tumors using a large series of families with mutations of the MLH1, MSH2, and MSH6 genes. DESIGN, SETTING, AND PARTICIPANTS: Families with Lynch syndrome enrolled between January 1, 2006, and December 31, 2009, from 40 French cancer genetics clinics participating in the ERISCAM (Estimation des Risques de Cancer chez les porteurs de mutation des genes MMR) study; 537 families with segregating mutated genes (248 with MLH1; 256 with MSH2; and 33 with MSH6) were analyzed. MAIN OUTCOME MEASURE: Age-specific cumulative cancer risks estimated using the genotype restricted likelihood (GRL) method accounting for ascertainment bias. RESULTS: Significant differences in estimated cumulative cancer risk were found between the 3 mutated genes (P = .01). The estimated cumulative risks of colorectal cancer by age 70 years were 41% (95% confidence intervals [CI], 25%-70%) for MLH1 mutation carriers, 48% (95% CI, 30%-77%) for MSH2, and 12% (95% CI, 8%-22%) for MSH6. For endometrial cancer, corresponding risks were 54% (95% CI, 20%-80%), 21% (95% CI, 8%-77%), and 16% (95% CI, 8%-32%). For ovarian cancer, they were 20% (95% CI, 1%-65%), 24% (95% CI, 3%-52%), and 1% (95% CI, 0%-3%). The estimated cumulative risks by age 40 years did not exceed 2% (95% CI, 0%-7%) for endometrial cancer nor 1% (95% CI, 0% 3%) for ovarian cancer, irrespective of the gene. The estimated lifetime risks for other tumor types did not exceed 3% with any of the gene mutations. CONCLUSIONS: MSH6 mutations are associated with markedly lower cancer risks than MLH1 or MSH2 mutations. Lifetime ovarian and endometrial cancer risks associated with MLH1 or MSH2 mutations were high but do not increase appreciably until after the age of 40 years. PMID- 21642683 TI - Long-term risks of subsequent primary neoplasms among survivors of childhood cancer. AB - CONTEXT: Survivors of childhood cancer are at excess risk of developing subsequent primary neoplasms but the long-term risks are uncertain. OBJECTIVES: To investigate long-term risks of subsequent primary neoplasms in survivors of childhood cancer, to identify the types that contribute most to long-term excess risk, and to identify subgroups of survivors at substantially increased risk of particular subsequent primary neoplasms that may require specific interventions. DESIGN, SETTING, AND PARTICIPANTS: British Childhood Cancer Survivor Study--a population-based cohort of 17,981 5-year survivors of childhood cancer diagnosed with cancer at younger than 15 years between 1940 and 1991 in Great Britain, followed up through December 2006. MAIN OUTCOME MEASURES: Standardized incidence ratios (SIRs), absolute excess risks (AERs), and cumulative incidence of subsequent primary neoplasms. RESULTS: After a median follow-up time of 24.3 years (mean = 25.6 years), 1354 subsequent primary neoplasms were ascertained; the most frequently observed being central nervous system (n = 344), nonmelanoma skin cancer (n = 278), digestive (n = 105), genitourinary (n = 100), breast (n = 97), and bone (n = 94). The overall SIR was 4 times more than expected (SIR, 3.9; 95% confidence interval [CI], 3.6-4.2; AER, 16.8 per 10,000 person-years). The AER at older than 40 years was highest for digestive and genitourinary subsequent primary neoplasms (AER, 5.9 [95% CI, 2.5-9.3]; and AER, 6.0 [95%CI, 2.3-9.6] per 10,000 person-years, respectively); 36% of the total AER was attributable to these 2 subsequent primary neoplasm sites. The cumulative incidence of colorectal cancer for survivors treated with direct abdominopelvic irradiation was 1.4% (95% CI, 0.7%-2.6%) by age 50 years, comparable with the 1.2% risk in individuals with at least 2 first-degree relatives affected by colorectal cancer. CONCLUSION: Among a cohort of British childhood cancer survivors, the greatest excess risk associated with subsequent primary neoplasms at older than 40 years was for digestive and genitourinary neoplasms. PMID- 21642684 TI - Characteristics of clinical trials to support approval of orphan vs nonorphan drugs for cancer. AB - CONTEXT: The Orphan Drug Act incentivizes medication development for rare diseases, offering substantial financial benefits to the manufacturer. Orphan products constitute most new drug approvals in oncology, but safety and efficacy questions have emerged about some of these agents. OBJECTIVES: To define characteristics of orphan cancer drugs and their pivotal clinical trials and to compare these with nonorphan drugs. DESIGN AND SETTING: We identified all new orphan and nonorphan drugs approved between 2004 and 2010 to treat cancer. We then collected data on key development variables from publicly available information on the US Food and Drug Administration's Web site and in the Code of Federal Regulations. MAIN OUTCOME MEASURES: We assessed clinical testing dates, approved indications, and regulatory characteristics (regular vs accelerated review, advisory committee review, postmarketing commitments). We then compared design features (randomization, blinding, primary end point) of pivotal trials supporting approval of orphan and nonorphan drugs and rates of adverse safety outcomes (deaths not attributed to disease progression, serious adverse events, dropouts) in pivotal trials. RESULTS: Fifteen orphan and 12 nonorphan drugs were approved between January 1, 2004, and December 31, 2010. Pivotal trials of orphan drugs had smaller participant numbers (median, 96 [interquartile range {IQR}, 66 152] vs 290 [IQR, 185-394] patients exposed to the drug; P < .001) and were less likely to be randomized (30% vs 80%; P = .007). Orphan and nonorphan pivotal trials varied in their blinding (P = .04), with orphan trials less likely to be double-blind (4% vs 33%). Primary study outcomes also varied (P = .04), with orphan trials more likely to assess disease response (68% vs 27%) rather than overall survival (8% vs 27%). More treated patients had serious adverse events in trials of orphan drugs vs trials of nonorphan drugs (48% vs 36%; odds ratio, 1.72; 95% confidence interval, 1.02-2.92; P = .04). CONCLUSION: Compared with pivotal trials used to approve nonorphan cancer drugs, pivotal trials for recently approved orphan drugs for cancer were more likely to be smaller and to use nonrandomized, unblinded trial designs and surrogate end points to assess efficacy. PMID- 21642686 TI - Association between time to initiation of adjuvant chemotherapy and survival in colorectal cancer: a systematic review and meta-analysis. AB - CONTEXT: Adjuvant chemotherapy (AC) improves survival among patients with resected colorectal cancer. However, the optimal timing from surgery to initiation of AC is unknown. OBJECTIVE: To determine the relationship between time to AC and survival outcomes via a systematic review and meta-analysis. data sources: MEDLINE (1975 through January 2011), EMBASE, the Cochrane Database of Systematic Reviews, and the Cochrane Central Register of Controlled Trials were searched to identify studies that described the relationship between time to AC and survival. STUDY SELECTION: Studies were only included if the relevant prognostic factors were adequately described and either comparative groups were balanced or results adjusted for these prognostic factors. DATA EXTRACTION: Hazard ratios (HRs) for overall survival and disease-free survival from each study were converted to a regression coefficient (beta) and standard error corresponding to a continuous representation per 4 weeks of time to AC. The adjusted beta from individual studies were combined using a fixed-effects model. Inverse variance (1/SE(2)) was used to weight individual studies. Publication bias was investigated using the trim and fill approach. RESULTS: We identified 10 eligible studies involving 15,410 patients (7 published articles, 3 abstracts). Nine of the studies were cohort or population based and 1 was a secondary analysis from a randomized trial of chemotherapy. Six studies reported time to AC as a binary variable and 4 as 3 or more categories. Meta-analysis demonstrated that a 4-week increase in time to AC was associated with a significant decrease in both overall survival (HR, 1.14; 95% confidence interval [CI], 1.10-1.17) and disease-free survival (HR, 1.14; 95% CI, 1.10-1.18). There was no significant heterogeneity among included studies. Results remained significant after adjustment for potential publication bias and when the analysis was repeated to exclude studies of largest weight. CONCLUSION: In a meta-analysis of the available literature on time to AC, longer time to AC was associated with worse survival among patients with resected colorectal cancer. PMID- 21642685 TI - KIT as a therapeutic target in metastatic melanoma. AB - CONTEXT: Some melanomas arising from acral, mucosal, and chronically sun-damaged sites harbor activating mutations and amplification of the type III transmembrane receptor tyrosine kinase KIT. We explored the effects of KIT inhibition using imatinib mesylate in this molecular subset of disease. OBJECTIVE: To assess clinical effects of imatinib mesylate in patients with melanoma harboring KIT alterations. DESIGN, SETTING, AND PATIENTS: A single-group, open-label, phase 2 trial at 1 community and 5 academic oncology centers in the United States of 295 patients with melanoma screened for the presence of KIT mutations and amplification between April 23, 2007, and April 16, 2010. A total of 51 cases with such alterations were identified and 28 of these patients were treated who had advanced unresectable melanoma arising from acral, mucosal, and chronically sun-damaged sites. INTERVENTION: Imatinib mesylate, 400 mg orally twice daily. MAIN OUTCOME MEASURES: Radiographic response, with secondary end points including time to progression, overall survival, and correlation of molecular alterations and clinical response. RESULTS: Two complete responses lasting 94 (ongoing) and 95 weeks, 2 durable partial responses lasting 53 and 89 (ongoing) weeks, and 2 transient partial responses lasting 12 and 18 weeks among the 25 evaluable patients were observed. The overall durable response rate was 16% (95% confidence interval [CI], 2%-30%), with a median time to progression of 12 weeks (interquartile range [IQR], 6-18 weeks; 95% CI, 11-18 weeks), and a median overall survival of 46.3 weeks (IQR, 28 weeks-not achieved; 95% CI, 28 weeks-not achieved). Response rate was better in cases with mutations affecting recurrent hotspots or with a mutant to wild-type allelic ratio of more than 1 (40% vs 0%, P = .05), indicating positive selection for the mutated allele. CONCLUSIONS: Among patients with advanced melanoma harboring KIT alterations, treatment with imatinib mesylate results in significant clinical responses in a subset of patients. Responses may be limited to tumors harboring KIT alterations of proven functional relevance. Trial Registration clinicaltrials.gov Identifier: NCT00470470. PMID- 21642687 TI - A strategic approach to therapeutic cancer vaccines in the 21st century. PMID- 21642688 TI - Balancing access and evaluation in the approval of new cancer drugs. PMID- 21642689 TI - Realigning incentives for developing and pricing new anticancer treatments. PMID- 21642690 TI - Seeing the future of cancer-associated transcription factor drug targets. PMID- 21642681 TI - Effect of screening on ovarian cancer mortality: the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Randomized Controlled Trial. AB - CONTEXT: Screening for ovarian cancer with cancer antigen 125 (CA-125) and transvaginal ultrasound has an unknown effect on mortality. OBJECTIVE: To evaluate the effect of screening for ovarian cancer on mortality in the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial. DESIGN, SETTING, AND PARTICIPANTS: Randomized controlled trial of 78,216 women aged 55 to 74 years assigned to undergo either annual screening (n = 39,105) or usual care (n = 39,111) at 10 screening centers across the United States between November 1993 and July 2001. Intervention The intervention group was offered annual screening with CA-125 for 6 years and transvaginal ultrasound for 4 years. Participants and their health care practitioners received the screening test results and managed evaluation of abnormal results. The usual care group was not offered annual screening with CA-125 for 6 years or transvaginal ultrasound but received their usual medical care. Participants were followed up for a maximum of 13 years (median [range], 12.4 years [10.9-13.0 years]) for cancer diagnoses and death until February 28, 2010. MAIN OUTCOME MEASURES: Mortality from ovarian cancer, including primary peritoneal and fallopian tube cancers. Secondary outcomes included ovarian cancer incidence and complications associated with screening examinations and diagnostic procedures. RESULTS: Ovarian cancer was diagnosed in 212 women (5.7 per 10,000 person-years) in the intervention group and 176 (4.7 per 10,000 person-years) in the usual care group (rate ratio [RR], 1.21; 95% confidence interval [CI], 0.99-1.48). There were 118 deaths caused by ovarian cancer (3.1 per 10,000 person-years) in the intervention group and 100 deaths (2.6 per 10,000 person-years) in the usual care group (mortality RR, 1.18; 95% CI, 0.82-1.71). Of 3285 women with false-positive results, 1080 underwent surgical follow-up; of whom, 163 women experienced at least 1 serious complication (15%). There were 2924 deaths due to other causes (excluding ovarian, colorectal, and lung cancer) (76.6 per 10,000 person-years) in the intervention group and 2914 deaths (76.2 per 10,000 person-years) in the usual care group (RR, 1.01; 95% CI, 0.96-1.06). CONCLUSIONS: Among women in the general US population, simultaneous screening with CA-125 and transvaginal ultrasound compared with usual care did not reduce ovarian cancer mortality. Diagnostic evaluation following a false-positive screening test result was associated with complications. Trial Registration clinicaltrials.gov Identifier: NCT00002540. PMID- 21642691 TI - Cancer risk assessment in Lynch syndrome: does the gene matter? PMID- 21642692 TI - JAMA patient page. Melanoma. PMID- 21642693 TI - Assessment of carotid atherosclerosis in normocholesterolemic individuals with proven mutations in the low-density lipoprotein receptor or apolipoprotein B genes. AB - BACKGROUND: Genetic cascade screening for heterozygous familial hypercholesterolemia (FH) revealed that 15% of individuals given this diagnosis do not exhibit elevated low-density lipoprotein cholesterol (LDL-C) levels. We assessed whether cardiovascular risk for these individuals differs from that of hypercholesterolemic FH heterozygotes and unaffected relatives. METHODS AND RESULTS: Individuals aged 18 to 55 years were recruited within 18 months after genetic screening. Three groups were studied: subjects given a molecular diagnosis of FH and with LDL-C levels at genetic screening below the 75th percentile (FH-low), subjects with FH and an LDL-C level above the 90th percentile (FH-high), and subjects without FH (no-FH). We measured carotid intima media thickness (IMT) by ultrasonography. Differences in carotid IMT among the groups were assessed using multivariate linear regression analyses. Mean carotid IMT of 114 subjects in the FH-low group (0.623 mm; 95% CI, 0.609 to 0.638 mm) was significantly smaller than that of 162 subjects in the FH-high group (0.664 mm; 95% CI, 0.648 to 0.679 mm; P<0.001) and did not significantly differ from the mean carotid IMT in 145 subjects in the no-FH group (0.628 mm; 95% CI, 0.613 to 0.642 mm; P=0.67). CONCLUSIONS: Our findings suggest that the risk of cardiovascular disease in patients with FH to a large extent is related to LDL-C levels and not to the presence of a mutation per se. Consequently, this study cautiously suggests that individuals with an FH genotype without expression of hypercholesterolemia may not require a pharmaceutical intervention that is as aggressive as the standard for subjects with FH. PMID- 21642704 TI - Rebuttal: Should we abandon the periodic health examination?: YES. PMID- 21642705 TI - Rebuttal: Should we abandon the periodic health examination?: NO. PMID- 21642706 TI - Identifying adults at risk of COPD who need confirmatory spirometry in primary care: Do symptom-based questions help? AB - Objective To examine the usefulness of a symptom-based case-finding questionnaire (CFQ) and the Medical Research Council (MRC) dyspnea scale in identifying which individuals with known risk factors for chronic obstructive pulmonary disease (COPD) require targeted spirometry in primary care. Design Cross-sectional study. Setting Three community primary care practices in Ontario. Participants Men and women 40 years of age and older with a smoking history of 20 pack-years or more. Main outcome measures We administered a CFQ for the presence of cough, sputum, wheeze, dyspnea, and recurrent respiratory infections (possible range of scores from 0 to 5) and applied the MRC dyspnea scale to assess the severity of COPD (possible range of scores from 1 to 5). Spirometric measures of forced expiratory volume in 1 second (FEV1) and forced vital capacity (FVC) were collected, with COPD defined as a postbronchodilator FEV1/FVC of less than 0.7 and FEV1 of less than 80% of the predicted value. Using spirometric data to confirm the diagnosis of COPD, likelihood ratios, pretest and posttest probabilities, and area under a receiver operating characteristic curve were calculated for the total CFQ and MRC scores. Results Scores for the CFQ and MRC dyspnea scale were available for 996 and 829 participants, respectively. The likelihood ratios for a total CFQ score of 3 or higher and an MRC dyspnea score of 4 or 5 were 1.82 (95% confidence interval [CI] 1.48 to 2.22) and 4.22 (95% CI 2.08 to 8.56), respectively. The likelihood ratios for a total CFQ score of 2 or less and an MRC dyspnea score of 1 were 0.75 (95% CI 0.66 to 0.85) and 0.50 (95% CI 0.39 to 0.65), respectively. Area under the receiver operating characteristic curve was 0.62 (95% CI 0.58 to 0.67; P < .001) for the total CFQ scores and 0.64 (95% CI 0.60 to 068; P < .001) for the MRC dyspnea scores. Conclusion In adults with known risk factors, the likelihood of having moderate to severe COPD is increased in those who report 3 or more common respiratory symptoms and marked functional limitation resulting from dyspnea. However, selecting individuals for spirometry based on symptoms alone will identify less than half of those with moderate to severe COPD. PMID- 21642708 TI - Accepting new patients: What does the public think about Ontario's policy? AB - Objective To gauge the public's opinion of the College of Physicians and Surgeons of Ontario's (CPSO's) policy on how primary care physicians should accept new patients. Design Deliberative citizens' council. Setting Toronto, Ont. Participants Twenty-five public members of the Toronto Health Policy Citizens' Council. Methods A 2-day council session was held, during which the new policy was presented and council members heard from experts with various perspectives on the issues involved. Council members then deliberated and developed recommendations concerning the policy. Main findings Council members agreed that a first-come, first-served policy was an appropriate method for family physicians to use when accepting new patients. They thought the policy's exception, which allows physicians not to accept patients based on a lack of clinical competency in an area, should be clarified in order to avoid it being used as an excuse to inappropriately screen patients. Counsel members also encouraged the CPSO to publicize its policy as widely as possible, so that potential patients undergoing screening in the future will recognize that this goes against the CPSO's policy and can take appropriate action if they wish. Conclusion How family physicians accept new patients into their practices is a sensitive issue. The CPSO policy provides guidance on how new patients should be admitted, which, if it is appropriately enacted, seems reasonable to informed members of the public. PMID- 21642707 TI - Prevalence of Bipolar Disorder symptoms in Primary Care (ProBiD-PC): A Canadian study. AB - Objective To describe the prevalence of patients who screen positive for symptoms of bipolar disorder in primary care practice using the validated Mood Disorders Questionnaire (MDQ). Design Prevalence survey. Setting Fifty-four primary care practices across Canada. Participants Adult patients presenting to their primary care practitioners for any cause and reporting, during the course of their visits, current or previous symptoms of depression, anxiety, substance use disorders, or attention deficit hyperactivity disorder. Main outcome measures Subjects were screened for symptoms suggestive of bipolar disorder using the MDQ. Health-related quality of life, functional impairment, and work productivity were evaluated using the 12-Item Short-Form Health Survey and Sheehan Disability Scale. Results A total of 1416 patients were approached to participate in this study, and 1304 completed the survey. Of these, 27.9% screened positive for symptoms of bipolar disorder. All 13 items of the MDQ were significantly associated with screening positive for bipolar disorder (P < .05). Patients screening positive were significantly more likely to report depression, anxiety, substance use, attention deficit hyperactivity disorder, family history of bipolar disorder, or suicide attempts than patients screening negative were (P < .001). Health-related quality of life, work or school productivity, and social and family functioning were all significantly worse in patients who screened positive (P < .001). Conclusion This prevalence survey suggests that more than a quarter of patients presenting to primary care with past or current psychiatric indices are at risk of bipolar disorder. Patients exhibiting a cluster of these symptoms should be further questioned on family history of bipolar disorder and suicide attempts, and selectively screened for symptoms suggestive of bipolar disorder using the quick and high-yielding MDQ. PMID- 21642709 TI - Effect of gender socialization on the presentation of depression among men: A pilot study. AB - Objective To examine the association between men's conformity to masculine norms and depression. Design Cross-sectional analysis. Setting University family practice clinic in Vancouver, BC. Participants Male patients, 19 years of age and older (N = 97). Main outcome measures The relationships among patients' scores on the Brief Symptom Inventory-18 depression subscale, Gotland Male Depression Scale, and Conformity to Masculine Norms Inventory, and whether or not patients were prompted to discuss emotional concerns with their physicians after completing these screening tests. Results Conformity to masculine norms was significantly associated with depression as assessed by the male depression screen (P = .039), but not with the screen that assessed typical depressive symptoms (P = .068). Men, regardless of their degree of masculinity or distress, overwhelmingly did not disclose emotional concerns to their physicians, even if the content of their distress involved suicidal thoughts. Conclusion Male depression screens might capture aspects of depression associated with masculine gender socialization that are not captured by typical measures of depression. Given the tendency of men to not disclose emotional distress to their family physicians, potentially high-risk cases could be missed without direct inquiry by clinicians. PMID- 21642710 TI - Le veritable mentorat en medecine. PMID- 21642711 TI - Le suicide des hommes. PMID- 21642712 TI - Too close for comfort: the effects of radical prostatectomy on intimacy: A professional couple's experience. PMID- 21642713 TI - Should we abandon the periodic health examination?: YES. PMID- 21642714 TI - Should we abandon the periodic health examination?: NO. PMID- 21642715 TI - Innocuousness of office-based olecranon bursa aspiration. PMID- 21642716 TI - Diclectin for morning sickness: Long-term neurodevelopment. AB - Question A pregnant patient recently asked me whether using Diclectin for morning sickness might affect the development of her child. Answer Our recent large study does show such a trend, although the differences are not necessarily clinically significant. PMID- 21642717 TI - Pharmacologic treatment of pediatric obesity. AB - Question There is a large population of overweight and obese children in my clinic. What medications for treatment of obesity are effective and can be used in children? Answer Orlistat is the only medication indicated by the US Food and Drug Administration for the treatment of obesity in adolescents. It is approved by the Food and Drug Administration for use in adolescents aged 12 years and older. There is no single approach to successful treatment of obesity, and lifestyle modification should be maintained throughout the pharmacologic treatment. PMID- 21642718 TI - "Can you go back to work?": Family physicians' experiences with assessing patients' functional ability to return to work. AB - Objective To explore the challenges academic FPs face when assessing patients' functional ability to return to work; to produce a detailed account of FPs' experiences and views on workplace disability management; to describe which parts of the disability assessment and management process FPs would like to modify or relinquish; and to provide solutions to streamline the overall process of assessing disability. Design Qualitative phenomenologic study using in-depth interviews. Setting A family health team located in a large urban teaching hospital in Toronto, Ont. Participants Purposive sample of 6 FPs. Methods Participants were invited to participate in 1-hour, in-depth interviews. Themes were derived from qualitative analysis of the data using a phenomenologic approach. Main findings Four themes emerged from the interviews: the FP's role in filing a compensation claim; the complexity of the patient; the FP's lack of training in occupational health; and possible solutions to improve the process of assessing the functional ability of an injured worker. Conclusion As in other areas of medicine, the role of the FP is to restore health; optimize social, psychological, and functional capabilities; and minimize the negative effects of injury. Assessing functional abilities for return to work can be challenging, as FPs are trained to focus on assessing and treating symptoms rather than on determining occupational functioning. Functional assessment forms do not provide enough information for physicians and serve as a poor communication tool among the stakeholders involved with returning an injured worker to work. PMID- 21642719 TI - David White MD CCFP (EM) FCFP. PMID- 21642732 TI - Family physicians' perspectives on personal health records: qualitative study. AB - OBJECTIVE: To explore FPs' perspectives on the value of personal health records (PHRs) in primary care and the implementation and adoption of PHRs in Canada. DESIGN: A qualitative design using semistructured interviews. SETTING: Southwestern Ontario. PARTICIPANTS: Ten FPs. METHODS: The 10 FPs participated in semistructured interviews, which were audiotaped and transcribed verbatim. An iterative approach using immersion and crystallization was employed for analysis. MAIN FINDINGS: Participants were generally positive about PHRs, and were attracted to their portability and potential to engage patients in health care. Their concerns focused on 3 main themes: data management, practice management, and the patient-physician relationship. Subthemes included security, privacy, reliability of data, workload, remuneration, physician obligations, patient misinterpretation of medical information, and electronic communication displacing face-to-face visits. Participants identified 3 key facilitators for adoption of PHR systems: integration with existing electronic health record systems, ease of use without being a burden on either time or money, and offering a demonstrated added value to family practice. CONCLUSION: This study replicates previously published literature about FP concerns and opinions, and it further identifies remuneration as a potential barrier in Canadian fee-for-service payment models. Participants identified 3 key facilitators, which were suggested for implementation and adoption of PHRs, providing a basis for future research and development of these systems for use in Canadian family practice. PMID- 21642733 TI - Family matters: does this mean the end of 24-hour on-call shift for Canadian residents? PMID- 21642735 TI - Clustering of opioid prescribing--what is really going on? PMID- 21642736 TI - Oxybutynin for treatment of nocturnal enuresis in children. AB - QUESTION: A 7-year-old child and his parents visit my clinic owing to the child's frequent bed-wetting. During the day, he has no problem controlling his urination. The family has tried behavioural methods but has failed to achieve dryness during the night. They ask to begin medical treatment. Is oxybutynin a safe and effective drug for treating nocturnal enuresis? ANSWER: Oxybutynin is an anticholinergic drug that has not been proven to be effective for treatment of nocturnal enuresis not accompanied by daytime symptoms, such as urgency. It can be added as a second-line drug and is effective for treating children with both daytime and nighttime wetting. Nevertheless, its common adverse effects, which can involve the central nervous system, should be considered when deciding whether or not to use it, especially in young children. PMID- 21642737 TI - Acute sinusitis. PMID- 21642738 TI - Navigating the complexity of ulcerative colitis: challenging case example. PMID- 21642739 TI - Tool to assess the quality of consultation and referral request letters in family medicine. PMID- 21642740 TI - A tale of two cultures: specialists and generalists sharing the load. AB - OBJECTIVE: To understand what contributes to good collaborative physician working relationships through identifying the factors that affect working relationships between generalist physicians and specialists. DESIGN: Qualitative study using in depth interviews. SETTING: University and community hospital inpatient family practice settings in Edmonton, Alta. PARTICIPANTS: Eleven physicians from various specialties who graduated between 1977 and 2001. METHODS: A grounded-theory approach, including constant comparison and creation of memorandums, helped to conceptualize the main concern and generated a framework for how the main concern was being resolved. A semistructured interview guide was developed and individual, in-depth interviews were audiotaped. Purposeful and theoretical sampling techniques were used. Three researchers participated in the analysis. MAIN FINDINGS: The findings suggested that when generalist physicians perceived that work had been imposed on them by specialists without negotiation, they felt overwhelmed by the workload. Differing priorities determined whether physicians were left holding the bag or sharing the load. In a system that valued technology and specialized knowledge and skills, the specialists were better able to control resources, set boundaries, and influence learners. This precipitated a culture of protecting valuable specialty resources, increasing physician isolation, and generalists feeling that they were left holding the bag. In order to reverse this cycle, it was important for physicians to develop good working relationships based on accessibility to needed expertise and tests, with negotiated agreements on how to share resources; mutual empowerment, including negotiation of roles and responsibilities to develop flexible relationships with a clear understanding of roles; and concern for fairness by sharing the load. CONCLUSION: Medical systems that value technology and focused interests might lead to someone being left holding the bag, contributing to generalists feeling overwhelmed and isolated within the system. A comprehensive system that values relationships might help to resolve issues created through perceived inequities in workload, disempowerment, and lack of understanding of roles. PMID- 21642744 TI - Aldo-keto reductase 1B7 is a target gene of FXR and regulates lipid and glucose homeostasis. AB - Aldo-keto reductase 1B7 (AKR1B7) is proposed to play a role in detoxification of by-products of lipid peroxidation. In this article, we show that activation of the nuclear receptor farnesoid X receptor (FXR) induces AKR1B7 expression in the liver and intestine, and reduces the levels of malondialdehyde (MDA), the end product of lipid peroxidation, in the intestine but not in the liver. To determine whether AKR1B7 regulates MDA levels in vivo, we overexpressed AKR1B7 in the liver. Overexpression of AKR1B7 in the liver had no effect on hepatic or plasma MDA levels. Interestingly, hepatic expression of AKR1B7 significantly lowered plasma glucose levels in both wild-type and diabetic db/db mice, which was associated with reduced hepatic gluconeogenesis. Hepatic expression of AKR1B7 also significantly lowered hepatic triglyceride and cholesterol levels in db/db mice. These data reveal a novel function for AKR1B7 in lipid and glucose metabolism and suggest that AKR1B7 may not play a role in detoxification of lipid peroxides in the liver. AKR1B7 may be a therapeutic target for treatment of fatty liver disease associated with diabetes mellitus. PMID- 21642745 TI - Plasma-based approach to measure target engagement for liver-targeting stearoyl CoA desaturase 1 inhibitors. AB - A positive correlation between stearoyl-CoA desaturase (SCD)1 expression and metabolic diseases has been reported in rodents and humans. These findings indicate that SCD1 is a promising therapeutic target for the chronic treatment of diabetes and dyslipidemia. The SCD1 enzyme is expressed at high levels in several human tissues and is required for the biosynthesis of monounsaturated fatty acids, which are involved in many biological processes. Liver-targeted SCD inhibitors were designed to pharmacologically manipulate SCD1 activity in the liver to avoid adverse events due to systemic inhibition. This article describes the development of a plasma-based SCD assay to assess the level of SCD inhibition, which is defined in this article as target engagement. Essentially, animals are dosed with an exogenous deuterated tracer (d7-stearic acid) as substrate, and the converted d7-oleic acid product is measured to monitor SCD1 inhibition. This study reveals that this plasma-based assay correlates with liver SCD1 inhibition and can thus have clinical utility. PMID- 21642746 TI - The social contagion effect of suicidal behavior in adolescents: does it really exist? AB - BACKGROUND: Suicide is the third leading cause of death among adolescents and a non-trivial percentage of adolescents report knowing someone who has attempted suicide. In light of this, a growing body of literature has explored whether suicidal behavior in one person may be imitated by others in their social networks. AIM: We seek to determine the extent to which suicidal behavior in individuals is influenced by suicidal behaviors of their peer and family members. METHODOLOGY: Using a nationally-representative sample of adolescents, we employ multivariate regression analysis with controls for known factors associated with suicidal behaviors to help isolate the effects of peer and family members on suicidal behaviors. Our methodology allows us to account for environmental confounders, simultaneity and to a limited extent, non-random peer selection. Our peer measures are drawn from the nomination of close friends by the individuals and suicidal behaviors among the peer group were constructed using the peers' own responses. RESULTS AND DISCUSSION: We find that a 10% increase in suicide attempts by family members were associated with a 2.13% and 1.23% increase in adolescent suicidal ideation and attempts, respectively. Our results also show that a 10% increase in peer suicidal ideation and attempts lead to a 0.7% and 0.3% increase in such behavior by the individuals. However, these positive associations between peer and individual suicide behavior become smaller and insignificant after adjustments were made for environmental confounders and peer selection. LIMITATIONS: Although we are able to establish the overall importance of environmental confounding factors, we are unable to identify the specific components or characteristics of the surroundings that can explain suicidality. The complex relationships between peer selection and suicidality also limit the determination of causality. CONCLUSIONS AND IMPLICATIONS: An increase in suicidal behavior by family members is positively associated with suicidal behavior among adolescents and effective policies aimed at reducing suicidal rates should consider these impacts. However, attributing correlations in suicidal behaviors among peers to social network effects should be undertaken with caution, especially when environmental confounders are not adequately controlled for in the analysis. FUTURE RESEARCH: Recent studies have found evidence that family connectedness and parent-child relationships have a significant impact in deterring risky behaviors among adolescents. This motivates future work aimed at designing policies that would utilize these findings in order to effectively reduce suicidal behavior among adolescents. PMID- 21642747 TI - Using referrals and priority-setting rules to risk adjust budgets: the case of regional psychiatric centers. AB - BACKGROUND: An important objective of many health care systems is to ensure equal access to health care services. One way of achieving this is by having universal coverage (low or absent out-of-pockets payments) combined with tax-financed transfers (block grants) to providers with a catchment area responsibility. However, a precondition for equal access in such systems is that providers have similar capacities -- meaning that budgets must be perfectly adjusted for variations in treatment costs not being under the control of providers (risk adjustment). AIM: This study presents a method that can be applied to adjust global budgets for variation in health risks. The method is flexible in the sense that it takes into account the possibility that variation in needs may depend on the degree of rationing in supplying health care services. METHODS: The information being available from referrals is used to risk-adjust budgets. An expert panel ranks each individual on the basis of need. The ranking is performed according to priority-setting criteria for health care services. In addition, the panel suggests an adequate treatment profile (treatment category and treatment intensity) for each referral reviewed. By coupling the treatment profiles with cost information, risk-adjusted budgets are derived. Only individuals found to have a sufficiently high ranking (degree of need) will impact the derived risk adjusted formula. RESULTS: The method is applied to four Regional Psychiatric Centers (RPC) supplying (i) outpatient services, (ii) day-patient care, and (iii) inpatient treatment for adults. The budget reallocations needed (positive and negative) to achieve an equal capacity across providers range between 10% and 42% of the current budgets. POLICY IMPLICATIONS: Our method can identify variations across providers when it comes to actual capacity and suggests budget reallocations that make the capacities to be equal across providers. In the case of the Regional Psychiatric Centers (RPCs) considered in this analysis, significant deviations in capacities are identified across providers and catchment areas. Thus, significant social gains can be gained, in terms of improved equal access, if our methodology is applied to risk adjust global budgets. PMID- 21642748 TI - The impact of mental illness on the risk of employment termination. AB - BACKGROUND: Mental illness can adversely impact labor market outcomes in a variety of ways, through education attainment, employment possibilities, and income. However, little is known about the impact of mental illness on an individual's ability to maintain employment. AIMS OF THE STUDY: This paper examines the impact of mental health on the risk of employment termination. We also distinguish between voluntary and involuntary employment termination. METHODS: Using data from the Medical Expenditure Panel Survey, we used survival analysis techniques to examine the impact of a mental illness diagnosis on the risk of employment termination and propensity score matching techniques to construct similar comparison groups. Initially, we used Cox proportional-hazards models where the event of interest was termination of employment. We then used a competing risk framework to differentiate between voluntary and involuntary employment termination. We also stratified our analysis by gender and capture levels of severity, receipt of treatment, and specific type of mental illness. RESULTS: We found that individuals with mental illness have an increased risk of employment termination in general as well as both involuntary and voluntary job loss. These results were consistent across all regression specifications. DISCUSSION: This paper is the first to explore the association between mental illness and the risk of employment termination in a sample of employed individuals. Employment is closely linked with mental health. Evidence suggests that employment can improve an individual's mental well-being but that job loss can have a detrimental effect. Our results suggest that individuals that can greatly benefit from their jobs are precisely those who are at high risk of losing them. IMPLICATIONS FOR HEALTH CARE PROVISION AND POLICIES: A greater effort must be made to provide support networks for employees within their work environments. IMPLICATIONS FOR HEALTH CARE RESEARCH: Future analysis should examine the impact of antidepressant therapy, supported employment policies, and other mental health-related interventions on the risk of employment termination for those individuals with mental illness. This would be instructive for advocates of the mentally ill to design more targeted interventions for individuals at risk. In addition, future research should examine the effect of other health conditions on the risk of employment termination. In the present study, poor self-reported health increased the risk of involuntary job termination. It would be informative to isolate what types of ailments contribute to this increased risk. PMID- 21642749 TI - Xenotropic murine leukemia virus-related virus (XMRV) in prostate cancer cells likely represents a laboratory artifact. AB - The prevalence of xenotropic murine leukemia virus-related virus (XMRV) in human population and its involvement in prostate cancer are subjects of ongoing research and debate. 22Rv1, which is a human cell line that serves as a common model of androgen-independent prostate cancer, was recently reported to carry infectious copies of XMRV. 22Rv1 was derived from a prostate cancer xenograft CWR22 that was serially passaged in immunodeficient mice. Based on the analysis of the DNA from CWR22 and 22Rv1, we present evidence against the presence of XMRV in CWR22 and, by inference, the tumor, from which CWR22 and 22Rv1 were established. While the presence of XMRV in 22Rv1 is likely to be an artifact, it may be a significant factor in determining the biological properties of this cell line. This consideration warrants additional caution for the interpretation of the relevance of the studies, which utilize this popular cell line as a model. It also invites a closer look at the sources of viral contamination in xenografts and cultured cells, as well as in the experiments that allege the presence of this virus in human cells and populations. PMID- 21642750 TI - First-principles atomistic study of surfaces of Fe-rich Fe-Cr. AB - The surface properties of Fe-rich ferromagnetic Fe-Cr alloys are investigated using a first-principles quantum-mechanical method. In dilute alloys, the surfaces are dominated by Fe, whereas the Cr-containing surfaces become favorable when the bulk Cr concentration exceeds the limit of ~ 10 atomic per cent. The abrupt change in the surface behavior is the consequence of complex competing magneto-chemical interactions between the alloying atoms. Considering the quantities of various features: equilibrium surface profiles, chemical potentials, segregation energies, surface energies, magnetic moments, mixing energies and pair interactions, within a wider range of bulk and surface concentrations enables us to build a comprehensive picture of the physics of Fe Cr surfaces. Using the present achievements many previously controversial results can now be merged into a consistent model of Fe-rich Fe-Cr alloys. PMID- 21642751 TI - Shear properties of potassium chloride films on iron obtained using density functional theory. AB - Density functional theory is used to calculate the shear strength of a thin KCl film grown epitaxially on an Fe(100) substrate. It is first demonstrated that the calculations accurately reproduce the experimental values of the shear moduli of bulk KCl. The method is then extended to calculating the shear properties of a three-layer slab of KCl on an Fe(100) substrate, where shear is found to take place at the KCl-Fe(100) interface and the KCl essentially moves as a rigid body. The resulting calculated values of the shear strength at zero pressure along the [linear span]10[linear span] (79.8 MPa) and [linear span]11[linear span] (70.3 MPa) directions are in excellent agreement with the experimentally measured value of 65 +/- 5 MPa. PMID- 21642752 TI - Nucleation and growth of Ag islands on the (?3 * ?3)R30 degrees phase of Ag on Si(111). AB - We use scanning tunneling microscopy to measure densities and characteristics of Ag islands that form on the (?3 * ?3)R30 degrees -Ag phase on Si(111), as a function of deposition temperature. Nucleation theory predicts that the logarithm of island density varies linearly with inverse deposition temperature. The data show two linear regimes. At 50-125 K, islands are relatively small, and island density decreases only slightly with increasing temperature. At 180-250 K, islands are larger and polycrystalline, and island density decreases strongly with increasing temperature. At 300 K, Ag atoms can travel for distances of the order of 1 um. Assuming that Ag diffusion occurs via thermally activated motion of single atoms between adjacent sites, the data can be explained as follows. At 50-125 K, the island density does not follow conventional Arrhenius scaling due to limited mobility and a consequent breakdown of the steady-state condition for the adatom density. At ~ 115-125 K, a transition to conventional Arrhenius scaling with critical nucleus size (i = 1) begins, and at 180-250 K, i > 1 prevails. The transition points indicate a diffusion barrier of 0.20-0.23 eV and a pairwise Ag-Ag bond strength of 0.14 eV. These energy values lead to an estimate of i~3-4 in the regime 180-250 K, where island density varies strongly with temperature. PMID- 21642753 TI - Island-size selectivity during 2D Ag island coarsening on Ag(111). AB - We report on the early stages of submonolayer Ag island coarsening on the Ag(111) surface carried out using kinetic Monte Carlo simulations for several temperatures. Our simulations were performed using a very large database of processes identified by their local environment and whose activation barriers were calculated using the semi-empirical interaction potentials based on the embedded-atom method. We find that during the early stages, coarsening proceeds as a sequence of selected island sizes, creating peaks and valleys in the island size distribution. This island-size selectivity is independent of initial conditions and results from the formation of kinetically stable islands for certain sizes as dictated by the relative energetics of edge atom detachment/attachment processes together with the large activation barrier for kink detachment. Our results indicate that by tuning the growth temperature it is possible to enhance the island-size selectivity. PMID- 21642754 TI - Surface alloy formation of noble adatoms adsorbed on Si(111)-?3 * ?3-Pb surface: a first-principles study. AB - The geometric structures, stability and electronic properties of initial stages of surface alloy formation for noble atoms adsorbed on Si(111)-)-?3 * ?3-Pb surfaces have been comparatively and extensively studied by using first principles calculations within density functional theory. Our results revealed that an Au trimer rather than a tetramer adsorption induces a surface alloy by combining with Pb atoms in covalent bonds, exhibiting semiconducting characteristics due to the localization of surface states. The stability of the two-dimensional (2D) surface alloy obeys the Hume-Rothery rule. The electronic structures of the 2D surface alloy are sensitive to the number of Au adatoms and can be modulated by the quantity of Au adatoms. Unlike the Au atoms, our further calculations indicated that adsorption of Ag or Cu atoms on the surface cannot form a surface alloy with Pb atoms in the surface layer due to a weaker interaction or smaller radius. PMID- 21642755 TI - Time-dependent density functional study of field emission from nanotubes composed of C, BN, SiC, Si, and GaN. AB - Field emission from various types of nanotubes is studied by propagating the electronic density in real space and time using time-dependent density functional theory. Capped (5, 5) C, BN, SiC, Si, and GaN nanotubes are considered. The GaN, SiC, and Si nanotubes were found to be significantly better field emitters than C and BN nanotubes, both in terms of current magnitude and sharpness of peaks in the energy spectra. By analyzing the electronic structure of the various systems it is seen that the nanotubes with the highest currents have electron densities that extend significantly from the nanotube in the emission direction. PMID- 21642756 TI - Magnetically responsive polypyrrole nanotubes using Ce(III)-stabilized maghemite nanoparticles. AB - Nanocomposites (NCs) that are made magnetically responsive in controlled conditions attract continuing interest for their added magnetic properties. In this study, we report on the preparation and full characterization of a multifunctional NC composed of magnetic gamma-Fe(2)O(3) nanoparticles (NPs) covalently attached to the surface of polyaminated (polyNH(2)) poly(2,6-di-pyrrol 1-yl-hexanoic acid) (pDPL) nanotubes (NTs). Such a hybrid conducting polymer iron oxide maghemite gamma-Fe(2)O(3)@pDPL NC built specifically on covalent bonding has never been reported. The maghemite gamma-Fe(2)O(3) NPs were prepared using an innovative ultrasound-assisted Ce(3+) doping process, resulting in polycarboxylation of the NP surface useful for control of aggregation and derivatization of functionality. The second component of the NC, i.e. polyNH(2) modified pDPL NTs, was prepared from an acid functional pyrrole species followed by amine modification. The resulting innovative gamma-Fe(2)O(3)@pDPL NC can be viewed as a multifunctional nanomaterial since it possesses both types of derivatization, i.e. polyCOOH (NPs) and polyNH(2) (NTs) combined with magnetic responsivity. PMID- 21642757 TI - Synthesis and mechanistic study of stable water-soluble noble metal nanostructures. AB - Sodium salt of poly(4-styrenesulfonic acid-co-maleic acid) (PSSMA) has been employed to prepare a series of stable nanosized metal colloids such as silver, gold, palladium, platinum, and silver-gold alloy nanostructures. All of the as synthesized products are very stable in water. The metal nanostructures have been directly confirmed by ultraviolet-visible spectroscopy, transmission electron microscopy (TEM), high-resolution TEM, and selected area electron diffraction (SAED), and also characterized by techniques such as Fourier transform infrared spectroscopy (FT-IR) and (1)H NMR. Intensive study has found that the metal ions are most probably reduced by organic radicals, generated from the thermal degradation of PSSMA. PMID- 21642758 TI - Controlled synthesis of gold nanoparticles by fluorescent light irradiation. AB - A novel photochemical synthesis of size-controlled gold nanoparticles was reliably accomplished via both a direct reduction and a seeded-growth method at room temperature under the irradiation of fluorescent light. These methods utilized the intensity of fluorescent light that closely resembles daily sunlight (~100 mW cm(-2)). This effectively allowed for the formation of gold nanoparticles with tunable sizes simply by controlling the concentration of trisodium citrate and gold chloride. The broad band fluorescent light was found to be an efficient source for inducing the formation of gold nanoparticles at ambient conditions. The size distribution and absorption property of the resulting nanoparticles were thoroughly characterized by scanning/transmission electron microscopy, dynamic light scattering, UV-visible spectroscopy and powder x-ray diffraction. This photochemical synthesis demonstrates, for the first time, the reliable preparation of gold nanoparticles at room temperature upon irradiation with fluorescent light. PMID- 21642759 TI - Structure and properties of nano-hydroxypatite scaffolds for bone tissue engineering with a selective laser sintering system. AB - In this study, nano-hydroxypatite (n-HAP) bone scaffolds are prepared by a homemade selective laser sintering (SLS) system based on rapid prototyping (RP) technology. The SLS system consists of a precise three-axis motion platform and a laser with its optical focusing device. The implementation of arbitrary complex movements based on the non-uniform rational B-Spline (NURBS) theory is realized in this system. The effects of the sintering processing parameters on the microstructure of n-HAP are tested with x-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM). The particles of n-HAP grow gradually and tend to become spherical-like from the initial needle-like shape, but still maintain a nanoscale structure at scanning speeds between 200 and 300 mm min(-1) when the laser power is 50 W, the light spot diameter 4 mm, and the layer thickness 0.3 mm. In addition, these changes do not result in decomposition of the n-HAP during the sintering process. The results suggest that the newly developed n-HAP scaffolds have the potential to serve as an excellent substrate in bone tissue engineering. PMID- 21642760 TI - Co-Pt nanoparticles encapsulated in carbon cages prepared by sonoelectrodeposition. AB - Co-Pt nanoparticles encapsulated in carbon cages have been prepared by sonoelectrodeposition followed by annealing in a CO atmosphere. Sonoelectrodeposition is a useful technique to make metallic nanoparticles, using ultrasound during electrodeposition to remove nanoparticles as they grow on the cathode surface. We used an electrolyte containing chloroplatinic acid and cobalt chloride and found that the atomic ratio of Co:Pt in the as-formed materials varied from 0.2 to 0.8 as the deposition current density was changed from 15 to 35 mA cm(-2). However, the as-deposited materials were inhomogeneous, comprising a mixture of Pt-rich and Co-rich nanoparticles. X-ray diffraction indicated that subsequent heat treatment (700 degrees C for 1 h) under CO gas created an ordered CoPt alloy phase that exhibited hard magnetic properties. Transmission electron microscopy showed many of the resulting nanoparticles to be encapsulated in carbon cages, which we ascribe to Co-catalyzed decomposition of CO during annealing. The thickness of the carbon cages was about ten layers, which may have helped reduce sintering during annealing. The size of the resultant nanoparticles was about 100 nm diameter, larger than the typical 5-10 nm diameter of as deposited nanoparticles. PMID- 21642761 TI - Crystalline Ge quantum dots embedded in SiO2 matrix synthesized by plasma immersion ion implantation. AB - A new plasma process, i.e. a combination of plasma immersion ion implantation and deposition (PIII&D) and high power impulse magnetron sputtering (HiPIMS), was developed to implant non-gaseous ions into material surfaces. The new process has the great advantage that thin film deposition and non-gaseous ion implantation can be achieved in a single plasma chamber. In this study, Ge ions were successfully implanted into SiO(2) thin film, which resulted in uniformly and homogeneously distributed crystalline Ge quantum dots (Ge-QDs) embedded in a SiO(2) matrix even without a further annealing process. Broader areas of application of PIII&D technology are envisaged with this newly developed process. PMID- 21642762 TI - Large-area pattern transfer of metallic nanostructures on glass substrates via interference lithography. AB - In this paper, we report a simple and effective nanofabrication method for the pattern transfer of metallic nanostructures over a large surface area on a glass substrate. Photoresist (PR) nano-patterns, defined by laser interference lithography, are used as template structures where a metal film of controlled thickness is directly deposited and then transferred onto a glass substrate by the sacrificial etching of the PR inter-layer. The laser interference lithography, capable of creating periodic nano-patterns with good control of their dimensions and shapes over a relatively large area, allows the wafer-scale pattern transfer of metallic nanostructures in a very convenient way. By using the approach, we have successfully fabricated on a glass substrate uniform arrays of hole, grating, and pillar patterns of Ti, Al, and Au in varying pattern periodicities (200 nm-1 um) over a surface area of up to several cm(2) with little mechanical crack and delamination. Such robust metallic nanostructures defined well on a transparent glass substrate with large pattern coverage will lead to advanced scientific and engineering applications such as microfluidics and nanophotonics. PMID- 21642763 TI - Appropriate use of diagnostic imaging in low back pain: a reminder that unnecessary imaging may do as much harm as good. AB - The rate of lumbar spine magnetic resonance imaging in the United States is growing at an alarming rate, despite evidence that it is not accompanied by improved patient outcomes. Overutilization of lumbar imaging in individuals with low back pain correlates with, and likely contributes to, a 2- to 3-fold increase in surgical rates over the last 10 years. Furthermore, a patient's knowledge of imaging abnormalities can actually decrease self-perception of health and may lead to fear-avoidance and catastrophizing behaviors that may predispose people to chronicity. The purpose of this clinical commentary is as follows: (1) to describe an outline of the appropriate use, as defined in recent guidelines, of diagnostic imaging in patients with low back pain; (2) to describe how inappropriate use of lumbar spine imaging can increase the risk of patient harm and contributes to the recent large increases in healthcare costs; (3) to provide physical therapists with clear guidelines to educate patients on both appropriate imaging and information to dampen the potential negative effects of imaging on patients' perceptions and health; and (4) to present an example of a successful clinical pathway that has reduced imaging and improved outcomes. LEVEL OF EVIDENCE: Diagnosis/prognosis/therapy, level 5. PMID- 21642765 TI - The implementation of international standardization of glycated hemoglobin. A "red-letter-day" for glycated hemoglobin in Italy: 1/1/11. Italian Recommendations of GLAD Working Group (A1c delegates WG). PMID- 21642764 TI - Effects of chronic slow release-lanreotide treatment on insulin-like growth factor system and metabolic parameters in acromegalic patients. AB - Insulin and IGF binding protein (IGFBP)-1 are linked by negative association. Somatostatin (SS) reduces insulin secretion by acting on pancreatic beta-cell and also by decreasing GH secretion. SS analogues in acromegaly reduce total IGF-I levels inhibiting GH hypersecretion, but they also reduce free IGF-I bioactivity increasing IGFBP-1 levels by inducing insulin decrease. In 13 acromegalic patients we studied GH, IGF system, insulin, and glucagon levels at baseline and at 7 days, 1 and 6 months under treatment with slow release (SR)-lanreotide (LAN) (60 mg im monthly). The hormonal and metabolic response to arginine (ARG) (0.5 g/kg iv in 30 min) was also studied at each time point. LAN decreased GH, total IGF-I, and IGFBP-3 levels at each time point. Insulin and glucagon levels were reduced, while IGFBP-1 and free IGF-I levels were increased by LAN at day 7 and after 1 month only. LAN did not modify the GH, insulin, glucagon, glucose, and IGFBP-1 responses to ARG. At each time point ARG-induced insulin increase was coupled to increase in glucagon and IGFBP-1 levels. This study shows that acromegalic patients under chronic treatment with LAN display: a) inhibition of GH and total IGF-I levels, not coupled to persistent decrease in free IGF-I levels; b) persistent decrease in IGFBP- 3 but transient decrease and increase in insulin and IGFBP- 1, respectively; c) unchanged hormonal and metabolic response to ARG. Our findings also show that ARG stimulates IGFBP-1 despite marked increase in insulin secretion; this escape from the negative relationship linking insulin and IGFBP- 1 would likely reflect the ARG-induced glucagon increase. PMID- 21642766 TI - Effects of maternal subclinical hypothyroidism on obstetrical outcomes during early pregnancy. AB - BACKGROUND: Maternal hypothyroidism [overt hypothyroidism and subclinical hypothyroidism (SCH)] during early pregnancy is suspected to associate with adverse obstetrical outcomes. AIM: The aim of the present study was to investigate whether maternal SCH during the early stage of pregnancy increase obstetrical complications and whether treatment results in an improvement in these outcomes. SUBJECTS AND METHODS: A total of 756 women in the 1st trimester (<=12 weeks) of pregnancy were enrolled through 10 hospitals in Shenyang from 2007 to 2009. All participants underwent thyroid function testing in early pregnancy and their obstetrical outcomes were studied following delivery. RESULTS: The incidence of spontaneous abortions in the SCH group was higher than the normal TSH group (15.48% vs 8.86%, p=0.03). No significant association was observed between SCH and other obstetrical complications including gestational hypertension, premature delivery, anemia, post-partum hemorrhage, low neonatal Apgar scores and low birth weight. Although levo-T4 (L-T4) treatment decreased the incidence of spontaneous abortions in women with SCH, it was not statistically significant when compared to women who did not receive treatment in the SCH group. None of the 28 women who received L-T4 treatment had premature delivery, low birth weight, hemorrhage, and low Apgar score. CONCLUSIONS: The incidence of spontaneous abortion in pregnant women with SCH increases in early pregnancy. PMID- 21642767 TI - Double membranes vs. lipid bilayers, and their significance for correct identification of macroautophagic structures. PMID- 21642768 TI - Antidepressants encounter autophagy in neural cells. AB - The prevailing view of antidepressants' (ADs) mode of action primarily focuses on their impact on neurotransmitter circuits, since the corresponding transporters and receptors are common targets of ADs. However, mounting evidence points to additional target structures, which may either support the beneficial effects or account for undesired side effects of ADs. Recently, we analyzed the influence of three ADs of different classes on autophagy-related processes in primary astrocytes and neurons. While amitriptyline (AMI) and citalopram (CIT) upregulate the expression of autophagic markers such as LC3B-II or Beclin 1, venlafaxine fails to exert these effects. Autophagy triggered by AMI and CIT is functional in terms of autophagic flux, and is partially mediated by class III PtdIns 3-kinase- and ROS dependent-pathways. Together, our study's results highlight a novel mode of action of ADs beyond monoaminergic neurotransmission. PMID- 21642770 TI - Is "X"-WAS the future for all of epidemiology? PMID- 21642769 TI - Simultaneous exposure of transformed cells to SRC family inhibitors and CHK1 inhibitors causes cell death. AB - The present studies were initiated to determine in greater molecular detail the regulation of CHK1 inhibitor lethality in transfected and infected breast cancer cells and using genetic models of transformed fibrobalsts. Multiple MEK1/2 inhibitors (PD184352, AZD6244 (ARRY-142886)) interacted with multiple CHK1 inhibitors (UCN-01 (7-hydroxystaurosporine), AZD7762) to kill mammary carcinoma cells and transformed fibroblasts. In transformed cells, CHK1 inhibitor -induced activation of ERK1/2 was dependent upon activation of SRC family non-receptor tyrosine kinases as judged by use of multiple SRC kinase inhibitors (PP2, Dasatinib; AZD0530), use of SRC/FYN/YES deleted transformed fibroblasts or by expression of dominant negative SRC. Cell killing by SRC family kinase inhibitors and CHK1 inhibitors was abolished in BAX/BAK -/- transformed fibroblasts and suppressed by over expression of BCL-XL. Treatment of cells with BCL-2/BCL-XL antagonists promoted SRC inhibitor + CHK1 inhibitor -induced lethality in a BAX/BAK-dependent fashion. Treatment of cells with [SRC + CHK1] inhibitors radio sensitized tumor cells. These findings argue that multiple inhibitors of the SRC RAS-MEK pathway interact with multiple CHK1 inhibitors to kill transformed cells. PMID- 21642771 TI - Random allocation in observational data: how small but robust effects could facilitate hypothesis-free causal inference. PMID- 21642772 TI - On standards of evidence. PMID- 21642774 TI - On the complex relationship between genes and environment in the etiology of autism. PMID- 21642775 TI - Autism spectrum disorders: is anything left for the environment? PMID- 21642776 TI - When implausible findings emanate from high-quality studies. PMID- 21642778 TI - Work as mental health-promoting or mental health-depreciating? PMID- 21642779 TI - Causal mediation analysis with survival data. PMID- 21642781 TI - Preregistration of epidemiology protocols. PMID- 21642782 TI - Cell phones and parotid cancer trends in England. PMID- 21642783 TI - Cannabis use and traffic injuries. PMID- 21642784 TI - Analgesics during pregnancy and cryptorchidism: additional analyses. PMID- 21642785 TI - A word and that to which it once referred: assessing "biologic" interaction. PMID- 21642786 TI - Ornithine transcarbamylase deficiency presenting as hepatitis. AB - Ornithine transcarbamylase deficiency is an inborn error of metabolism that commonly presents as hyperammonemia in neonates. We present a case of a 2-year old girl who was referred to a pediatric emergency department for evaluation of hepatitis, an uncommon presentation of ornithine transcarbamylase deficiency. Recognition of late presentations of this disease is important for survival and neurological outcome. PMID- 21642787 TI - A case of autonomically mediated pain due to spinal epidural abscess in an adolescent female. AB - Spinal epidural abscess (SEA) is a rare disease in children without predisposing risk factors. Atypical presentations of SEA without the classic triad of fever, back pain, and neurological signs, have previously been described. We report an atypical presentation of an acute SEA in a previously healthy adolescent female. This 15-year-old patient presented with right shoulder pain in the absence of the fever, back pain, or long-tract signs; therefore, the diagnosis of the spinal pathology was delayed. Eventually, a thoracic SEA was identified by gadolinium enhanced magnetic resonance imaging and treated with surgical decompression followed by intravenously administered antibiotics. The patient's course was complicated by chronic headache. Our experience adds to the literature a case that demonstrates the difficulty in diagnosis of atypical presentations of SEA in previously healthy children. In addition, referred or autonomically mediated pain should be considered in unusual pain presentations. Children with significant extremity or abdominal pain should be considered for the possibility of sympathetically mediated pain syndrome due to a thoracic-level spinal cord lesion such as a SEA. PMID- 21642788 TI - Cholelithiasis in a toddler with sickle cell disease. AB - Cholelithiasis is rarely seen in toddlers and school-aged children, even in the setting of sickle cell anemia. In addition to more common etiologies, such as gastroenteritis, constipation, and urinary tract infection, the differential diagnoses of acute abdominal pain in young children with sickle cell disease include vaso-occlusive pain crisis and splenic sequestration. We describe a case of a toddler with sickle cell disease initially presenting with abdominal pain who was found to have symptomatic cholelithiasis. PMID- 21642789 TI - Late migration of a foreign body in the foot 5 years after initial trauma. AB - An 11-year-old boy presented to our emergency department because of the extrusion of a small piece of glass through the plantar skin, 5 years after the initial trauma. Another foreign body was identified on ultrasounds and removed surgically. Ultrasound is an accurate test for detection of foreign bodies and to assess potential complications. The treatment of all foreign bodies is surgical exploration and removal, except if the risks of attempting removal are higher than the potential damage of the foreign body in its existing location. Migrations of foreign bodies are not uncommon. Ultrasounds could help for localization and removal as an intraoperative procedure. In case of suspicion of a foreign body after trauma, complementary examinations are required. PMID- 21642790 TI - Rhabdomyolysis without detectable myoglobulinuria due to severe hypophosphatemia in diabetic ketoacidosis. AB - Clinical signs of hypophosphatemia, even when severe, are rare in diabetic ketoacidosis despite their high frequency in this condition. This article presents a patient with rhabdomyolysis due to severe hypophosphatemia, where the level of serum phosphorus was observed to be as low as 0.42 mg/dL on the 16th hour of ketoacidosis treatment. The patient developed acute tubular necrosis due to rhabdomyolysis, but there was no blood reaction in the urine, and the creatine kinase increased to 1200 U/L. The patient was treated without dialysis and was cured after a polyuria period of 2 months after the oliguric period. PMID- 21642791 TI - First-Onset Seizure After Use of an Energy Drink [corrected]. AB - The health consequences of energy drink use in adolescents are unknown. We discuss an adverse event in an adolescent who presented to the emergency department with his first-ever seizure after consumption of 5-Hour Energy. We review the typical presentation of caffeine toxicity, as well as the importance of screening for energy drink use in adolescents with appropriate clinical findings. We pay particular attention to the identification of energy drink related adverse events in the emergency department and the need for subsequent reporting to the Food and Drug Administration. To our knowledge, this is the first reported case of an adolescent presenting with a new-onset seizure associated with energy drink use. PMID- 21642792 TI - Successful intravenous thrombolysis in a 14-year-old boy with ischemic stroke. AB - We report a healthy 14-year-old boy with an acute left middle cerebral artery stroke, treated 2 hours after the onset of symptoms with intravenous recombinant tissue plasminogen activator (r-TPA). Recanalization of the middle cerebral artery was documented with transcranial Doppler during the first 5 minutes of intravenous r-TPA perfusion, and progressive recovery of the neurological deficits occurred. Although lack of evidence regarding safety and efficacy in children precludes the recommendation of systematic use of r-TPA in pediatric stroke, we propose that this option should be considered and discussed with the parents, especially in older children presenting within 3 hours in centers with experience in adult thrombolysis. PMID- 21642793 TI - Globe rupture and nonaccidental trauma: two case reports. AB - The association of globe rupture and nonaccidental trauma is not established in the literature. We present 2 cases involving a 9-month-old infant girl and a 14 month-old girl who experienced inflicted traumatic globe rupture. In the first case, the infant had isolated eye findings on physical examination, but extensive injuries were revealed on the skeletal survey and computed tomography of the head. In the second case, the toddler had a prior history of abuse, so our index of suspicion for nonaccidental trauma was heightened. Despite isolated eye findings on physical examination, this toddler also had evidence of additional injury on skeletal survey. These cases illustrate the importance of considering nonaccidental trauma in infants and toddlers who present with globe rupture and performing a complete evaluation for child abuse if no history is given or if the history is inconsistent with the injury. PMID- 21642794 TI - Staphylococcal infection mimicking child abuse: what is the differential diagnosis and appropriate evaluation? AB - Twins with similar skin lesions are described. Although initially concerning for nonaccidental burn injury, further evaluation led to the diagnosis of bullous impetigo caused by Staphylococcus aureus. Thoughtful assessment is important in such cases to protect the child and prevent misdiagnosis. PMID- 21642795 TI - Case records of the Children's Mercy Hospital: A 12-year-old girl with a straddle injury. AB - Straddle injuries are common in children. At the end of this case presentation, you should be able to describe the approach to the evaluation and treatment of a straddle injury, list indications for gynecologic consultation and/or sedation, plan disposition, and discuss pitfalls to avoid in evaluating patients with straddle injuries. PMID- 21642796 TI - Diaphragmatic avulsion with chance fracture: a rare association in the seat belt syndrome. AB - Traumatic diaphragmatic injuries are uncommon in children. Chance fractures are typical in adults who had motor vehicle accidents but infrequent in children. The seat belt syndrome is characterized by a spectrum of injuries caused by the compression of the abdomen by the seat belt. It can affect the abdominal wall, hollow or solid organs, and the spine, but the association between diaphragmatic rupture and Chance fracture is extremely rare. We describe the case of a child who was involved in a car accident and who presented with multiple traumas; liver and lung injuries were first detected, and the patient was managed accordingly. During the hospital stay, a right diaphragmatic avulsion and a Chance fracture, which were initially missed, were diagnosed and treated successfully. The relevance of this case lies in the low incidence of the diaphragmatic rupture and the Chance fracture in children along with their exceptional association and in the diagnosis, which might be especially difficult in the acute setting. PMID- 21642797 TI - Peripherally inserted central catheters. AB - Peripherally inserted central catheters are increasingly used in the pediatric and adolescent population for long-term central access. This article reviews the indications, insertion techniques, and complications of peripherally inserted central catheter lines. PMID- 21642799 TI - Hospital-based pandemic influenza preparedness and response: strategies to increase surge capacity. AB - In the spring of 2009, the first patients infected with 2009 H1N1 virus were arriving for care in hospitals in the United States. Anticipating a second wave of infection, our hospital leaders initiated multidisciplinary planning activities to prepare to increase capacity by expansion of emergency department (ED) and inpatient functional space and redeployment of medical personnel. EXPERIENCE: During the fall pandemic surge, this urban, tertiary-care children's hospital experienced a 48% increase in ED visits and a 12% increase in daily peak inpatient census. However, several strategies were effective in mitigating the pandemic's impact including using a portion of the hospital's lobby for ED waiting, using a subspecialty clinic and a 24-hour short stay unit to care for ED patients, and using physicians not board certified in pediatric emergency medicine and inpatient-unit medical nurses to care for ED patients. The average time patients waited to be seen by an ED physician and the proportion of children leaving the ED without being seen by a physician was less than for the period when seasonal influenza peaked in the winter of 2008-2009. Furthermore, the ED did not go on divert status, no elective medical or surgical admissions required cancellation, and there were no increases in serious patient safety events. SUMMARY: Our health center successfully met the challenges posed by the 2009 H1N1 outbreak. The intent in sharing the details of our planning and experience is to allow others to determine which elements of this planning might be adapted for managing a surge of patients in their setting. PMID- 21642800 TI - ECGs in the ED. PMID- 21642801 TI - The price of perfection: a teenaged athlete with elevated serum creatinine. PMID- 21642802 TI - Illustrative case history: neurofibromatosis. PMID- 21642803 TI - Measurement of carboxyhemoglobin. PMID- 21642804 TI - Setting a research agenda for simulation-based healthcare education: a synthesis of the outcome from an Utstein style meeting. PMID- 21642805 TI - Author conflict and bias in research: quantifying the downgrade in methodology. PMID- 21642806 TI - Evidence-based recommendations for spine surgery. PMID- 21642808 TI - Re: Halpern CH, Milby AH, Wensheng G, et al. Clearance of the cervical spine in clinically nonevaluable trauma patients. Spine 2010;35:1721-8. PMID- 21642809 TI - Re: Halpern CH, Milby AH, Wensheng G, et al. Clearance of the cervical spine in clinically nonevaluable trauma patients. Spine 2010;35:1721-8. PMID- 21642810 TI - Efficacy of a brief screening tool to identify medical students in distress. AB - PURPOSE: To determine whether the Medical Student Well-Being Index (MSWBI) can serve as a brief assessment tool to identify medical students in severe psychological distress. METHOD: The authors used data from 2,248 medical students at seven U.S. medical schools who responded to a 2007 survey to explore the accuracy of the MSWBI in identifying medical students with three outcomes: low mental quality of life (QOL; defined by having a Medical Outcomes Study Short Form Health Survey mental component summary score >=1/2 standard deviation below that of the age- and gender-matched population norm), suicidal ideation, or serious thoughts of dropping out. The authors confirmed their analyses using data from a separate sample of 2,682 students evaluated in 2009. RESULTS: Students with low mental QOL, suicidal ideation, or serious thoughts of dropping out were more likely to endorse each individual MSWBI item and a greater number of total items than were students without such distress (all P < .001). The likelihood ratio for low mental QOL among students with MSWBI scores <4 was 0.47 as compared with 4.79 for those with scores >=4. At an MSWBI threshold score of >=4, the MSWBI's sensitivity and specificity for identifying students with low mental QOL or recent suicidal ideation/serious thoughts of dropping out were both >=90%. On multivariable logistic regression, all MSWBI items were independently associated with at least one outcome. CONCLUSIONS: The MSWBI is a useful brief screening tool to help identify students with severe distress. PMID- 21642811 TI - Recognition memory and beta-amyloid plaques in adult Tg2576 mice are not modified after oral exposure to aluminum. AB - The role of aluminum (Al) in Alzheimer disease is highly controversial. However, this element has been detected in neuritic plaques and neurofibrillary tangles in patients with Alzheimer disease. Its presence in neuritic plaques in hippocampus is especially relevant, as this is an area closely related to spatial learning and memory. In this study, the diet of wild-type and Tg2576 mice (animals overexpressing the human amyloid precursor protein) was supplemented with Al lactate (1 mg/g). General neurotoxic Al effects were evaluated using a functional observational battery and a novel object recognition task. Four experimental groups were used: Control-wild, Al-wild, Control-Tg, and Al-Tg mice. The results show a decreased home-cage activity and an increase in piloerection in all Al exposed animals, and an increased sensorimotor reactivity in Tg2576 mice given Al. Neither Al treatment nor genotype had any noticeable effect on corticosterone levels and Al concentrations in frontal cortex and cerebellum of the mice. Recognition memory was impaired in Tg2576 mice, whereas beta-amyloid plaque depositions were observed in all these animals. However, Al did not alter the recognition memory and beta-amyloid plaque loads of Tg2576 mice. PMID- 21642812 TI - Polycythemia from mast cell activation syndrome: lessons learned. AB - A middle-aged woman presented with fatigue and mild increases in hematocrit and red cell mass. Polycythemia vera was diagnosed. She underwent therapeutic phlebotomy but clinically worsened. On reevaluation, other problems were noted including episodic malaise, nausea, rash and vasomotor issues. The JAK2V617F mutation was absent; paraneoplastic erythrocytosis was investigated. Serum tryptase and urinary N-methylhistamine were normal, but urinary prostaglandin D2 was elevated. Skin and marrow biopsies showed no mast cell abnormalities. Extensive other evaluation was negative. Gastrointestinal tract biopsies were histologically normal but revealed increased, aberrant mast cells on immunohistochemistry; the KITD816V mutation was absent. Mast cell activation syndrome, recently identified as a clonal disorder involving assorted KIT mutations, was diagnosed. Imatinib 200 mg/d rapidly effected complete, sustained response. Diagnosis of mast cell activation syndrome is hindered by multiple factors, but existing therapies for mast cell disease are usually achieve significant benefit, highlighting the importance of early diagnosis. Multiple important aspects of clinical reasoning are illustrated by the case. PMID- 21642813 TI - Delayed allergic reaction after coronary angiography. AB - Contrast-enhanced angiography is a very useful tool for the diagnosis and evaluation of vascular diseases. Contrast media (CM) were first introduced in the 1930s and since then they have become an important diagnostic method. Nonetheless, using CM have their price, as some patients may develop adverse reactions to them. These reactions to CM are divided into chemotoxic and allergic/pseudoallergic. Both reactions range in their time of appearance, clinical manifestations, severity, pathomechanisim, diagnostic methods and the way they are managed. Late adverse reactions to CM may be easily overlooked as they are less reported. In the present case report, the authors present a patient who developed a delayed response to nonionic CM used during an elective coronary angiography, followed by a review of the current literature and suggested recommendations. PMID- 21642814 TI - Rhabdomyolysis in a 48-year-old man with hepatitis B-induced cirrhosis. AB - Telbivudine (beta-l-2'-deoxythymidine) is an orally administered nucleoside analog drug approved for the treatment of patients with chronic hepatitis B since 2006. Initially, it was regarded as being generally well tolerated, with low adverse reaction profiles and no dose-limiting toxicity. Recently, with the result of the telbivudine worldwide phase III GLOBE trial and other clinical experiments, cases of myopathy and neuropathy have been reported undergoing long term telbivudine treatments. Telbivudine-induced myopathy has been characterized by muscle pain, weakness and moderately elevated creatine kinase levels during treatments, although no severe adverse events have been observed. Rhabdomyolysis has not reported in any patient. PMID- 21642815 TI - Acute cystic rupture and hemorrhagic shock after a vigorous massage chair session in a patient with polycystic kidney disease. AB - Although cyst bleeding is a common complication in polycystic kidney disease (PCKD), cyst rupture is uncommon even in the setting of trauma. The risk factors, morbidity and mortality of cystic rupture in patients with PCKD are not well established. The authors herein present a case of perinephric hemorrhage secondary to cyst rupture in a 77-year-old man with PCKD after a vigorous massage chair session and provide a thorough review of related cases in the literature. PMID- 21642816 TI - Pitfalls with radiopharmaceuticals. AB - INTRODUCTION: There is a considerable body of evidence describing that the pharmacokinetics and pharmacodynamics of radiopharmaceuticals may be changed by a variety of drugs, disease states and in some cases, surgical procedures. OBJECTIVE: : To systematically search the medical literature and review the published evidence on adverse reactions to radiopharmaceuticals. METHOD: MEDLINE, EMBASE, International Pharmaceutical Abstracts and Science Citation Index were searched for studies reporting adverse reactions to radiopharmaceuticals. Controlled trials, cohort studies, case-control studies and case series published in major Western languages were considered for the review. Each study included in the present review was described in a narrative way, and major components of each study were reported (ie, research design, patient characteristics, types of drugs and radiopharmaceuticals, dosing information and adverse reactions). RESULTS: The majority of adverse reactions to radiopharmaceuticals described in the literature required little or no treatment, and their negative effects were generally mild and self-limited. Large longitudinal greater than 5-year studies reported prevalence rates of adverse reactions due to radiopharmaceuticals ranging from 0 to 25 cases per 100,000 administrations. Case studies on the use of technetium reported mild adverse reactions; however, some led to potentially harmful complications. Similarly, studies involving fluorodeoxyglucose reported more severe adverse reactions. CONCLUSION: The literature on radiopharmaceuticals adverse effects is scarce, and just a few studies were conducted to investigate the association between radiopharmaceuticals and adverse reactions. Despite relatively mild and self-limited symptoms, the current widespread use of radiopharmaceuticals requires constant monitoring for adverse reactions. PMID- 21642817 TI - Does long-term aspirin use have any effect on Helicobacter pylori eradication? AB - INTRODUCTION: Antimicrobial resistance has decreased eradication rates for Helicobacter pylori infection, and recent reports from different countries report eradication rates lower than 80% with triple therapy. The aim of this pilot study was to investigate the efficacy of standard triple eradication regimen in long term aspirin users. METHOD: The study population consisted of 77 aspirin using patients with dyspeptic symptoms and 79 age- and sex-matched dyspeptic patients without aspirin use as a control group. Both the study group and control patients were given lansoprazole (30 mg twice a day), clarithromycin (500 mg twice a day) and amoxicillin (1 g twice a day) (LCA) for 14 days as the eradication regimen. Patients on the study group were allowed to take aspirin during the eradication regimen (LCAAsp). Eradication was defined as the absence of H pylori as assessed with the C-urea breath test and H pylori stool antigen test 8 weeks after the end of the antimicrobial therapy. RESULTS: The H pylori eradication rate in the LCAAsp group was 64/77 [83%, 95% confidence interval (CI): 79%-94%] with intention-to-treat (ITT) analysis and 64/75 (85%, 95% CI: 82%-96%) with per protocol (PP) analysis, and the H pylori eradication rate in the LCA group was 42/79 (53%, 95% CI: 43%-65%) with ITT analysis and 42/75 (56%, 95% CI: 46%-68%) with PP analysis. The difference between the groups both with ITT analysis and with PP analysis was statistically significant (P < 0.05). CONCLUSION: These data suggest that H pylori eradication rate with standard triple eradication regimen is significantly higher among long-term aspirin users than in controls. PMID- 21642818 TI - Deep vein thrombosis, inferior vena cava interruption and multiple thrombophilic gene mutations. AB - Interruption or hypoplasia of the inferior vena cava, with associated azygos continuation, is an uncommon congenital vascular malformation (Ellis et al, Comput Radiol 1986;10:15-22). Although this anomaly causes venous stasis, few patients present with history of deep vein thrombosis (DVT). The exact role of coexisting thrombophilic gene mutations, also heterozygotic, is far from being completely understood. However, in these cases, because of a probable additive effect, treatment of complications and careful prophylaxis for recurrent DVT are recommended lifelong. The authors report a case of inferior vena cava interruption with azygos continuation in a 30-year-old woman who presented with a history of recurrent lower limb DVT. In addition, heterozygosis for the H1299R polymorphism of the factor V gene (Factor V HR2), for the C677T polymorphism of the methylenetetrahydrofolate reductase gene (MTHFR C677T) and for the 4G/5G polymorphism of the plasminogen activator inhibitor-1 gene (PAI-1 4G/5G) was found in DNA analyses. PMID- 21642819 TI - Tinnitus is associated with increase in the intima-media thickness of carotid arteries. AB - INTRODUCTION: Increased intima-media thickness of carotid arteries is considered as an early indicator of atherosclerosis. The authors here aimed to assess whether tinnitus is associated with such thickening by a cross-sectional approach. METHODS: A total of 820 male (53.5 +/- 9.6 years) and 528 female (54.5 +/- 9.7 years) Japanese undergoing a brain screening program including ultrasonographical evaluation of carotid arteries between April 2007 and April 2009 were included in the study. Increased intima-media thickness was defined as >= 1 mm. Multiple logistic regression analyses were performed with adjustment for sex, age, body mass index, hypertension and smoking. RESULTS: Increased intima media thickness was observed in 53.1% overall (in 57.6% of men and in 46.2% of women), with significant associations with tinnitus (odds ratio, 1.62; 95% confidence interval, 1.19-2.22), age, male gender and hypertension. The tinnitus link persisted after adjustment for the confounding factors. CONCLUSIONS: Tinnitus is associated with increased intima-media thickness of carotid arteries in Japanese undergoing a brain screening program. PMID- 21642820 TI - Transplantation of human umbilical cord blood mesenchymal stem cells improves survival rates in a rat model of acute hepatic necrosis. AB - INTRODUCTION: Stem cell-based therapies are emerging as important and promising methods in the treatment of end-stage liver disease. This study is aimed to evaluate the effects of human umbilical cord blood mesenchymal stem cell (HUCBMSC) transplantation in acute hepatic necrosis (AHN). METHODS: Green fluorescent protein (GFP)-labeled HUCBMSCs were injected into the liver of rats in which AHN was induced by carbon tetrachloride, and the migration of these cells in liver slices was evaluated from 48 hours to 4 weeks post transplantation. The transdifferentiation status of the HUCBMSCs was evaluated using immunohistochemistry and real-time reverse transcription-polymerase chain reaction, and survival rates were statistically analyzed. RESULTS: Dispersed GFP fluorescence was observed along the portal area 48 hours after transplantation. One week post-transplantation, GFP-positive cells were found in necrotic liver areas, and GFP-positive cells persisted after 4 weeks. Immunohistochemistry and real-time polymerase chain reaction analysis showed that transplanted HUCBMSCs expressed several human liver tissue-specific markers in rats with AHN. Statistical analysis revealed that rats with AHN that were transplanted with HUCBMSCs had significantly lower death rates after 48 hours than those receiving no HUCBMSCs. CONCLUSION: HUCBMSC transplantation can significantly improve the survival of rats with AHN. The underlying mechanisms involved may include the transdifferentiation of HUCBMSCs into hepatocyte-like cells and targeted migration of these cells to liver lesion sites. PMID- 21642821 TI - Endoplasmic reticulum stress caused by left ventricular hypertrophy in rats: effects of telmisartan. AB - INTRODUCTION: Studies have revealed that excessive endoplasmic reticulum (ER) stress leads to apoptosis. Although cardiomyocytes apoptosis contributes to the transition from left ventricular hypertrophy (LVH) to heart failure, it is unknown whether ER stress participates in the pathologic process. The authors first induced coarctation of the abdominal aorta in rats to induce LVH and then investigated the effect of telmisartan on the resulting ER stress. METHODS: Male Sprague-Dawley rats were randomly divided into 3 groups: sham operation, abdominal aortic coarctation (AAC) and AAC + telmisartan. Telmisartan (5 mg . kg . d) or vehicle was infused into the stomach 1 week after the operation. ER stress signaling pathway molecules and apoptosis were studied in pressure overloaded hearts 9 weeks after AAC. RESULTS: Telmisartan significantly reduced LVH and interstitial fibrosis and improved left ventricular function compared with AAC alone. Cardiac markers of ER stress such as GRP78, C/EBP homologous protein, caspase-12 and phospho c-Jun NH2-terminal kinase were significantly increased in rats with AAC, and telmisartan significantly blunted these changes. Rats that received both telmisartan and AAC had less apoptosis due to ER stress. CONCLUSIONS: Increased ER stress might be responsible for enhanced cardiomyocyte apoptosis after aortic coarctation. Telmisartan may reduce ER stress and thereby attenuate both apoptosis and cardiac hypertrophy. PMID- 21642822 TI - A primary care focus on the treatment of patients with major depressive disorder. AB - Major depressive disorder (MDD) is a common psychiatric illness affecting nearly 20% of adults in the United States at least once during their lifetime. MDD is frequently diagnosed and treated in the primary care setting. Management of the disease may be complicated by patients and family members feeling stigmatized by the diagnosis and not understanding that depression is a treatable medical illness, which, in turn, fosters low rates of adherence to treatment recommendations. Incomplete or delayed response to treatment, adverse events associated with antidepressants and medical or psychiatric comorbidities also interfere with optimal depression management. This article presents an overview of diagnostic and treatment guidelines for MDD and focuses on challenges encountered by primary care physicians. The role of antidepressant medications, psychotherapy and nonpharmacologic interventions for the treatment of patients with MDD is described, and factors influencing treatment selection, such as adverse event profiles and patient characteristics, are examined. PMID- 21642823 TI - Polymerase chain reaction-based detection of Pneumocystis jirovecii in bronchoalveolar lavage fluid for the diagnosis of pneumocystis pneumonia. AB - INTRODUCTION: The diagnosis of pneumocystis pneumonia (PCP) in non-human immunodeficiency virus (HIV)-infected immunocompromised patients is notoriously difficult. The recent advent of polymerase chain reaction (PCR)-based detection systems, based on the identification of single fungal genes, has markedly improved diagnostic accuracy in this ominous disease. In an attempt to further improve diagnostic yield, the authors used a PCR-based detection system for Pneumocystis jirovecii, based on targeting 3 distinct genes. METHODS: During the 4-year period (January 2005 to January 2009), all consecutive immunocompromised patients suspected of having PCP in the differential diagnosis underwent bronchoscopy with bronchoalveolar lavage sampling for the evaluation of the etiology of pulmonary infiltrates. Bronchoalveolar fluid was tested for the presence of a wide variety of possible etiological microorganisms. RESULTS: In a cohort of 214 immunocompromised patients (of which 198 were non-HIV immunocompromised patients) who underwent bronchoscopy with bronchoalveolar lavage for evaluation of pulmonary infiltrates, PCR correctly diagnosed PCP in 75% (42/56) compared with 14% (8/56) diagnosed by traditional stains, and increased diagnostic yield 5.4-fold. CONCLUSIONS: Given the absence of a sensitive gold standard, this study demonstrates the usefulness of a multigene PCR-based detection of Pneumocystis jirovecii DNA for supporting the clinical diagnosis of PCP, with high sensitivity and negative predictive value rates compared with direct stains, especially in non-HIV immunocompromised patients. PMID- 21642824 TI - Acute limb ischemia. AB - Acute limb ischemia is a medical emergency with management options ranging from urgent revascularization to limb amputation. The best patient outcome requires tailoring the treatment to the individual patient. This article describes a step by-step approach for diagnosis and management of patients presenting with acute limb ischemia. PMID- 21642825 TI - Deep infiltrating cervical endometriosis mimicking rectosigmoid cancer. PMID- 21642826 TI - Acute liver failure: an uncommon complication of commonly used medication. AB - The incidence of drug-related acute liver failure is approximately 14 per 100,000 populations. Drug-induced liver injury may take place through a variety of mechanism. Withdrawal of the offending agent may result in complete recovery. Clindamycin is known to cause mild derangement of liver function; however, acute liver injury causing severe derangement of liver function associated with encephalopathy is uncommon. PMID- 21642827 TI - On the differences between ouabain and digitalis glycosides. AB - Digoxin and digitoxin are widely used in the treatment of heart diseases. The exact mechanism of action of these drugs has remained an enigma. Ouabain has become the standard tool to investigate the mode of action of cardiotonic steroids, and results with ouabain are regarded as generally valid for all cardiac glycosides. However, there are marked differences between the effects of ouabain and digitalis glycosides. Ouabain has a different therapeutic profile from digitalis derivatives. Unlike digitalis glycosides, ouabain has a fast onset of action and stimulates myocardial metabolism. The inotropic effect of cardiotonic steroids is not related to inhibition of the Na-K-ATPase. Ouabain and digitalis derivatives develop their effects in different cellular spaces. Digitalis glycosides increase the intracellular calcium concentration by entering the cell interior and acting on the ryanodine receptors and by forming transmembrane calcium channels. Ouabain, by activation of the Na-K-ATPase from the extracellular side, triggers release of calcium from intracellular stores via signal transduction pathways and activates myocardial metabolism. These data no longer support the concept that all cardiotonic steroids exhibit their therapeutic effects by partial inhibition of the ion-pumping function of the Na-K ATPase. Hence, it is suggested that this deeply rooted dogma be revised. PMID- 21642828 TI - Implications of over-the-counter proton pump inhibitors for patient counseling by pharmacists. AB - The article reviews the literature on the role of the pharmacist in patient counseling and discusses how that role may apply to patients with frequent heartburn who are seeking an over-the-counter (OTC) treatment. Searches of the National Library of Medicine PubMed database were conducted using the terms "heartburn," "nonprescription," "therapy," "pharmacist," and "counseling," supplemented by additional searches on counseling for prescription products, and by the author's knowledge of pharmacy practice and the scientific literature. Accurate recognition of the signs of heartburn are an important first step in counseling a patient on the appropriate OTC medication; immediate referral to a health care provider is mandatory if cardiac pain or certain gastrointestinal symptoms are present. When counseling a patient about treatments for heartburn, the pharmacist should practice effective listening in an environment that is conducive to communication by the patient. Proton pump inhibitors are effective for the treatment of heartburn; the histamine2 receptor antagonists and antacids should also be considered for appropriate patients. Adverse events have been noted with proton pump inhibitors; however, overall the benefits significantly outweigh the risks and problems are unlikely to arise during the 2-week duration of OTC treatment of heartburn. Pharmacists can provide valuable services to patients with frequent heartburn, particularly with regard to counseling about the condition and appropriate OTC therapy. The availability of numerous OTC products, including antacids, histamine2 receptor antagonists, and proton pump inhibitors, enables pharmacists to fulfill an important clinical role and improve patient satisfaction. PMID- 21642829 TI - Emergency use of intravenous phytonadione (vitamin K1) for treatment of severe bleeding in a child with chronic cholestasis. AB - We present a 5-year-old boy with multiple hematomas associated with chronic cholestasis. A week before admission he suffered minor trauma at day care. The next day he complained of trunk and limb pain and orthopedic consultation, including leg x-rays, revealed no abnormalities. Over the next 5 days multiple hematomas developed over his body and increased in size. In the Emergency Department he was in pain and looked sick but alert. He had fever and tachycardia, with normal blood pressure and respiratory status and physical examination showed several hematomas on the legs, which increased in size during observation in the Emergency Department over 2 hours. Blood work revealed multiple coagulation abnormalities, and International Normalized Ratio was 12. Intravenous phytonadione (vitamin K1) was immediately administered with normalization of coagulation abnormalities within 1 hour and the hematomas stopped growing in size. In addition to missing follow-up with the Pediatric Gastroenterology Department, social service agency inquiry found he had not taken his medications for several months. With severe abnormal bleeding and hepatic disease, intravenous vitamin K1 may be lifesaving, even before obtaining confirmatory blood work, fresh-frozen plasma, or blood transfusion. PMID- 21642830 TI - Symptomatic-low grade methemoglobinemia because of dapsone: a multiple hit hypothesis. AB - Methemoglobinemia occurs when hemoglobin is oxidized to form methemoglobin (MetHb) rendering it incapable of oxygen transport and leading to tissue hypoxia. Typically, protective mechanisms keep MetHb levels below 1%, but a variety of compounds are capable of inducing methemoglobinemia including dapsone. Normally, symptoms correlate with MetHb level and treatment with methylene blue is reserved for patients with significantly elevated MetHb levels. We present a case of symptomatic low-grade methemoglobinemia because of dapsone therapy. This case illustrates a multiple hit hypothesis where comorbid conditions prompt symptoms at a low MetHb level. Understanding the impact of comorbid conditions in exacerbating low-grade methemoglobinemia is important in diagnosing and managing this condition. Treatment with methylene blue should be guided by the whole clinical picture rather than by the MetHb level alone. PMID- 21642831 TI - Therapeutic options for obstructive sleep apnea. AB - Obstructive sleep apnea is a common, chronic disorder characterized by the cessation or reduction in airflow due to periodic mechanical obstruction of the upper airway passage during sleep. Symptoms and signs of sleep apnea include daytime sleepiness, fatigue, poor concentration, snoring, resuscitative snorts, and related occupational accidents. Sleep apnea is classified into 3 types based on its severity and management varies accordingly. The first step in treatment is to identify correctible causes and educate patients on lifestyle modifications. Positive airway pressure is the preferred treatment modality and can be delivered via 3 different modes. Oral appliances are of 3 types and are generally indicated in mild to moderate obstructive sleep apnea. Surgery is a final therapeutic option that focuses on the removal of excessive tissue from different sites of the pharyngeal airway. PMID- 21642832 TI - Vitamin D and Blood Pressure Homeostasis. PMID- 21642833 TI - Alternative agents to vancomycin for the treatment of methicillin-resistant Staphylococcus aureus infections. AB - Resistant gram-positive infections, specifically methicillin-resistant Staphylococcus aureus (MRSA), carry an increased risk for morbidity and mortality. Historically, MRSA has been a cause of nosocomial infections, although recent reports have noted an increased prevalence in community-acquired MRSA infections. Vancomycin is the preferred agent to treat MRSA. However, cases of S. aureus with reduced susceptibility to vancomycin have been reported, prompting the need for alternative treatment options. In this review, we discuss the currently available agents with MRSA activity and those in development. Linezolid and quinupristin/dalfopristin have been demonstrated as effective although potential toxicities must be taken into consideration before their use. Daptomycin, tigecycline, telavancin, and ceftaroline are well tolerated but lack the clinical data to support a superior place in treatment over vancomycin. Several new agents in various stages of development have also demonstrated MRSA activity. Currently, vancomycin remains the gold-standard treatment option for MRSA infections. In situations that limit its use, consideration of patient specific parameters, cost, and relevant clinical data demonstrating drug safety and efficacy should be employed for the selection of the appropriate alternative agent. PMID- 21642834 TI - Does seating location impact voting behavior on Food and Drug Administration advisory committees? AB - Food and Drug Administration (FDA) advisory committees have a significant influence on patterns of clinical practice worldwide. Recent guidance to the committees by the FDA has focused on attempting to eliminate sources of bias due to committee voting procedures. Nevertheless, major sources of social influence have not been addressed. We analyzed transcripts of Circulatory Systems Devices Panel meetings from 1997 to 2005 in which the panel cast votes on premarket approval and for which a voting minority existed. Committee members who are assigned to speak later are significantly more likely to be in the voting minority (P < 0.001). This effect holds for meetings with sequential voting (P = 0.0058) and for meetings with simultaneous voting (P = 0.045). A weaker effect shows that, for meetings with sequential voting, committee members who vote later are significantly more likely to be in the voting minority (P = 0.018). Speaking order and voting order are both determined by seating location. We therefore conclude that voting behavior on FDA expert advisory committees is strongly associated with seating location. This suggests the presence of a possible social dynamic that is not addressed by existing FDA committee procedures. PMID- 21642835 TI - Pulmonary arterial hypertension: classification and therapy with a focus on prostaglandin analogs. AB - Pulmonary arterial hypertension, part of the larger spectrum of disorders causing pulmonary hypertension, is a complex and progressive disease of multiple etiologies that ultimately leads to vascular remodeling, right-sided heart failure, and death. Advances in treatment over the past 15 to 20 years have dramatically reduced the morbidity and mortality of the disease, but often have significant drawbacks. Of the more recently approved therapies, the prostaglandin analogs have been shown to have the greatest therapeutic benefit but are also the most difficult to administer, many being given as continuous intravenous infusions in the ambulatory setting. After a case presentation highlighting some of the challenges that accompany treatment with these agents, this article reviews the diagnosis and classification of pulmonary hypertension and pulmonary arterial hypertension and gives a brief overview of the various other pharmacologic agents used in its treatment. A more comprehensive review of the biochemistry of prostaglandins and the pharmacology and clinical use of this class of drugs follows. Recommended treatment guidelines are also discussed. PMID- 21642836 TI - Effect of carvedilol versus metoprolol CR/XL on mortality in patients with heart failure treated with cardiac resynchronization therapy: a COX multivariate regression analysis. AB - We investigated in 136 consecutive patients with heart failure receiving cardiac resynchronization therapy (CRT) the effect of carvedilol versus metoprolol CR/XL versus no beta blocker on mortality. Of the 136 patients, 42 (31%) were on carvedilol, 80 (59%) were on metoprolol CR/XL, and 14 (10%) were not on a beta blocker. A decrease of left ventricular end-systolic volume >=15% after CRT was defined as a positive response to CRT. Of the 136 patients, 62 (46%) responded to CRT. It was found that both carvedilol and metoprolol CR/XL were not related to CRT response on using Cox univariate regression analysis. Twenty-two of the 136 patients (16%) died during follow-up of 17 +/- 10 months after initiating CRT. Mortality occurred in 14 of 80 patients (18%) on metoprolol CR/XL, in 3 of 42 patients (7%) on carvedilol, and in 5 of 14 patients (36%) not on beta blockers (P = 0.04). After adjustment for age, gender, and the variables with significant differences by Cox univariate regression, both carvedilol (hazard ratio = 0.14; P = 0.03; 95% confidence interval = 0.02-0.86) and metoprolol CR/XL (hazard ratio = 0.19; P = 0.02; 95% confidence interval = 0.04-0.80) were found to be related to mortality by Cox multivariate regression. PMID- 21642837 TI - Effect of a photoactivated rhodium complex in melanoma. AB - cis-Dichlorobis (3,4,7,8-tetramethyl-1,10-phenanthroline) rhodium(III) chloride (OCTBP) is an octahedral complex that has been shown to react with nucleic acids when irradiated with light. Earlier studies on its phototoxicity toward human esophageal, bladder, pancreas, and colon cancer cells have been extended to in vitro and in-vivo evaluation of its effect on malignant melanoma cells using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H tetrazolium proliferation assay and xenograft model of melanoma. Tumor volume and the percentage of tumor growth delay were used to assess the antitumor effect of light-activated OCTBP. Terminal deoxynucleotidyl transferase dUTP nick-end labeling assays and immunohistochemistry were also performed on tumor tissue samples. Photoactivated OCTBP was found to inhibit melanoma cell growth by 40% at 100 MUmol/l concentration. Phototherapy with OCTBP was most efficacious with optimal percentage treated/control values observed early in the course of the experiment. At the end of the experiment (60 days), there was a 50% tumor inhibition compared with vehicle or drug controls. OCTBP treatment resulted in a tumor growth delay of 16 days (tumor growth of 24%) compared with the untreated control, whereas cisplatin resulted in 10 days tumor growth delay. This antitumor activity was found to be closely associated with the induction of apoptosis. The use of photoactivated OCTBP as an adjuvant therapy for the treatment of melanoma may provide a new targeted therapy to prevent progression of this disease. PMID- 21642838 TI - Predominant enhancement of apoptosis induced by methyl jasmonate in bladder cancer cells: therapeutic effect of the Antp-conjugated Smac peptide. AB - Methyl jasmonate (MJ) has recently attracted attention as a promising antitumoral compound because of its highly specific proapoptotic properties in a wide range of malignancies. However, the high doses required to achieve a therapeutic benefit have limited its clinical development. Here, we hypothesize that the family of inhibitor of apoptosis proteins (IAPs) may inhibit MJ-mediated apoptosis in cancer cells. We combined MJ with the IAPs inhibitor, the second mitochondria-derived activator of caspases (Smac) peptide to treat bladder cancer cells. The results showed that the combination of MJ and Smac peptide enhanced the apoptosis-inducing effect in a synergistic manner by releasing and activating IAPs-bounding caspase-3. These findings suggest that the inhibition of IAPs could overcome the resistance of cancer cells to MJ. PMID- 21642839 TI - Long-term response in advanced bladder cancer involving the use of temsirolimus and vinflunine after platin resistance. AB - Relapse after initial first-line chemotherapy shows a poor prognosis in metastatic urothelial cancer. Currently, several chemotherapeutic agents and targeted drugs are under evaluation for platin-resistant advanced urothelial carcinoma. Vinflunine has been approved for second-line treatment in this indication. We present a patient with initial T4 advanced and subsequently metastasized bladder cancer, who has shown prolonged survival of 44 months after radical cystectomy. During her clinical course, the patient received two different platinum-containing therapies, temsirolimus within a phase II protocol and subsequent vinflunine chemotherapy. Treatment duration was 15 weeks with temsirolimus and 9 weeks with vinflunine, respectively, with a stable disease period of 3.8 months under temsirolimus therapy. This case is an example of how patients can derive a survival benefit from adequate sequencing of surgery and medical treatment including the newest therapies, even in advanced disease. PMID- 21642840 TI - Triterpenoid pristimerin synergizes with taxol to induce cervical cancer cell death through reactive oxygen species-mediated mitochondrial dysfunction. AB - A combined treatment with conventional chemotherapies can enhance the effectiveness of chemotherapeutic agents against cancers. Here, we have shown that the naturally occurring triterpenoids synergistically enhance the response of cervical cancer cells to taxol. Of the triterpenoid compounds, pristimerin enhanced the anticancer effect of taxol with the highest efficiency by combination. Pristimerin synergizes with taxol to inhibit clonogenic survival and tumor growth in nude mice, and to enhance cell death in cervical cancer cells. A combined treatment with taxol and pristimerin induced cervical cancer cell death by increasing intracellular reactive oxygen species levels, upregulation of death receptor death receptor 5 (DR5), activation of Bax, and dissipation of mitochondrial membrane potential. Treatment with N-acetyl-L-cysteine, a thiol containing antioxidant completely blocked combined treatment-induced Bax translocation as well as DR5 upregulation. Moreover, inhibition of Jun N-terminal kinase/c-Jun pathway attenuated cell death by blocking DR5 upregulation and Bax activation. These results indicate that the triterpenoid, pristimerin, synergistically enhances taxol response of cervical cancer cells through DR5 expression and Bax activation. Furthermore, the reactive oxygen species-dependent activation of the Jun N-terminal kinase/c-Jun pathway is required for the DR5 upregulation and Bax activation. The molecular mechanism revealed by this study may aid in the design of future combination cancer therapies against cells with intrinsically reduced sensitivity to taxol. PMID- 21642841 TI - Transient, three-dimensional flow field simulation through a mechanical, trileaflet heart valve prosthesis. AB - Thromboembolic complications are one of the major challenges faced by designers and researchers in development of artificial organs with blood-contacting devices such as heart valve prostheses, especially mechanical valves. Besides increasing the thrombogenic potential, these valves change the hydrodynamic performance of the heart. In this study, the flow through a trileaflet, mechanical heart valve prosthesis was modeled with transient computational fluid dynamics to analyze flow patterns causing thrombus formations on valves. The valve was simulated under conditions of a test rig (THIA II), which was specially designed to analyze different valves with respect to thrombosis. The main goal of this study was to mimic the exact conditions of the test rig to be able to compare numerical and experimental results. The boundary conditions were obtained from experimental data as leaflet kinematics and pressure profiles. One complete cycle of the valve was simulated. Numerical flow and pressure results were analyzed and compared with experimental results. Shear stress and shear rates were determined with respect to thrombogenic potential, especially in the pivot regions, which seem to be the main influence for activation and deposition of thrombocytes. Approximately 0.7% of the blood volume moving through the fluid domain of the valve was exposed to shear rates high enough to cause platelet activation. However, shear rates of up to 20,000 s-1 occurred in pivot regions. The pressure differences between the simulation and experimental data were approximately 2.5% during systole and increased up to 25% during diastole. The presented method, however, can be used to gain more information about the flow through different heart valve prostheses and, thus, improve the development process. PMID- 21642842 TI - Ethical considerations for ventricular assist device support: a 10-point model. AB - The potential for long-term support on a ventricular assist device (VAD) in the bridge-to-transplant (BTT) and destination therapy (DT) settings has created unprecedented ethical challenges for patients and caregivers. Concerns include the patient's adaptation to life on a device and the ethical, clinical, and practical issues associated with living on mechanical support. On the basis of our experience treating 175 consecutive VAD patients, we have developed a model to address the ethical and psychosocial needs of patients undergoing VAD implantation. Patient preparation for VAD implantation encompasses three phases: 1) initial information regarding the physical events involved in implantation, risks and benefits of current device technology, and the use of VAD as a rescue device; 2) preimplant preparation including completion of advance directives specific to BTT/DT, competency determination, and identifying a patient spokesperson, multidisciplinary consultants, and cultural preferences regarding device withdrawal; and 3) VAD-specific end-of-life issues including plans for device replacement and palliative care with hospice or device withdrawal. This three-phase 10-point model addresses the ethical and psychosocial issues that should be discussed with patients undergoing VAD support. PMID- 21642843 TI - Successful use of temporary right ventricular support to avoid implantation of biventricular long-term assist device: a transcutaneous approach. AB - Severe right ventricular (RV) failure after left ventricular assist device (LVAD) implantation is a serious complication with a poor prognosis. The most effective therapy for these patients is an upgrade to biventricular mechanical support. However, it is well recognized that primary and secondary biventricular ventricular assist device implantation is associated with higher mortality rates. We report on three patients with cardiogenic shock, who were provided on an emergency basis with a percutaneous extracorporeal life support (ECLS) system by the femoral vessels. After stabilization, a LVAD was implanted. To avoid secondary RV failure, the ECLS was switched to a transcutaneous RV assist device (RVAD) as a temporary RV support. The arterial cannula was removed from the femoral artery, and a prosthesis-supported cannula was sutured to the pulmonary artery and passed through a subxiphoid exit. The femoral vein cannula was left in situ. Both cannulae were connected to a centrifugal pump. Two patients could be weaned from the RVAD; the system was explanted under local anesthesia. One patient died due to internal bleeding. In conclusion, the ease of device implantation, weaning, and explantation justifies a liberal use of temporary RV support to avoid implantation of the problematic long-term biventricular assist devices. PMID- 21642844 TI - Changes in pain-related beliefs, coping, and catastrophizing predict changes in pain intensity, pain interference, and psychological functioning in individuals with myotonic muscular dystrophy and facioscapulohumeral dystrophy. AB - OBJECTIVES: The primary aim of this study was to test hypothesized associations between changes in psychological variables (ie, pain beliefs, catastrophizing, and coping strategies) and changes in pain intensity and related adjustment (ie, pain interference and psychological functioning) in individuals with myotonic muscular dystrophy (MMD) and facioscapulohumeral muscular dystrophy (FSHD). METHODS: A sample of 107 adults with a diagnosis of MMD or FSHD, reporting pain in the past 3 months, completed assessments at 2 time points, separated by approximately 24 months. RESULTS: Results showed that changes in pain-related psychological variables were significantly associated with changes in psychological functioning, pain intensity, and pain interference. Specifically, increases in the belief that emotion influences pain, and catastrophizing were associated with decreases in psychological functioning. Increases in the coping strategies of asking for assistance and resting, and the increases of catastrophizing were associated with increases in pain intensity. Finally, increases in pain intensity and asking for assistance were associated with increases in pain interference. DISCUSSION: The results support the use of the biopsychosocial model of pain for understanding pain and its impact in individuals with MMD or FSHD. These findings may inform the design and implementation of psychosocial pain treatments for people with muscular dystrophy and chronic pain. PMID- 21642845 TI - Treatment of chronic low back pain: a randomized clinical trial comparing multidisciplinary group-based rehabilitation program and oral drug treatment with oral drug treatment alone. AB - OBJECTIVE: This randomized clinical trial examined the efficacies of a group based multidisciplinary rehabilitation program and oral drug treatment versus oral drug treatment alone in Iran. METHODS: A total of 197 patients with chronic low back pain were randomized to either intervention group (n=97) receiving a group-based, 5-session multidisciplinary rehabilitation program plus oral medication or to control group (n=100) receiving just oral medication. At baseline and at 3 and 6-month follow-ups, patients filled out questionnaires on health-related quality of life (36-item Short-form General Health Survey) and disability Questionnaires (Quebec Disability Scale and Ronald-Morris Disability). Repeated measure analysis of variance was used to compare 2 groups during time. RESULTS: The 2 groups were comparable regarding all baseline characteristics (P>0.05). There were significant differences within each group by time in terms of all subscales of 36-item Short-form (P<0.01) except for mental health (P=0.7). Furthermore, there were significant differences between groups in terms of all domains of SF-36 scale except for general health (P=0.06), social function (P=0.08) and role emotional (P=0.7). Furthermore, according to the scores of Ronald-Morris Disability Questionnaire and Quebec Disability Scale, the disability of patients in the intervention group was improved over time significantly (P=0.01 and P<0.0001, respectively). DISCUSSION: The findings revealed that the group-based multidisciplinary program could improve most domains of quality of life in chronic low back pain patients in the 6-month period. However, there were no significant differences between two groups in sub scales such as general health, social function and role emotional. PMID- 21642846 TI - Pain-associated mild sensory deficits without hyperalgesia in chronic non neuropathic pain. AB - OBJECTIVES: A mixture of sensory loss and gain is a hallmark of neuropathic pain. But hypesthesia and hyperalgesia also occur with experimentally induced acute pain. Here, we assessed sensory profiles in chronic non-neuropathic pain (osteoarthritis, OA) using the quantitative sensory testing (QST) protocol of the German Research Network on Neuropathic Pain (DFNS). METHODS: Twenty individuals with OA [mean pain intensity on the numerical rating scale (NRS, 0-10): 5.6+/ 1.5] were tested on the painful and contralateral hand and compared with 20 healthy volunteers matched for age, sex, and handedness. RESULTS: In the OA group, analysis of variance revealed increased detection thresholds to tactile stimuli bilaterally and to thermal stimuli restricted to the more painful hand (all P<0.05). Pin-prick hypoalgesia was present restricted to the patients' more affected hand. Neither hyperalgesia nor allodynia was found. QST parameters were correlated with average pain intensity (r between 0.48 and 0.51). CONCLUSIONS: These results suggest that chronic non-neuropathic pain may induce slight sensory impairment for large fiber function (bilateral) and small fiber function (ipsilateral). However, all changes are within the normal range, in contrast to patients with neuropathy. Inhibition of central pathways by nociceptive input and altered sensory processing due to disuse of the hand are possible mechanisms. These functional sensory alterations do not interfere with the diagnosis of neuropathy. PMID- 21642847 TI - Variables associated with decreasing pain among persons living with human immunodeficiency virus: a longitudinal follow-up study. AB - BACKGROUND: Pain is common among persons with human immunodeficiency virus (HIV); however, there are minimal data on its natural history, or the long-term efficacy of analgesic therapies. METHODS: We performed an observational study between 2001 and 2009. Pain was defined on a 0 to 10 scale; 0=no pain; 10=worst pain possible. Patients were included if they were HIV positive, had a chronic pain diagnosis, a median pain score during the first year of observation of >=1.0, >=2 years of follow-up, and >=3 recorded pain scores. Two models were used to describe decreasing pain. Model 1 defined decreasing pain as a negative slope to the best fit line through all recorded pain scores. Model 2 defined decreasing pain as a median pain score of zero during the last year of follow-up. RESULTS: Using model 1, decreasing pain was negatively associated with a history of being abused (odds ratio=0.29) and positively associated with peripheral neuropathy (3.54). Using model 2, decreasing pain was positively associated with highly active antiretroviral therapy (3.71) and negatively associated with opioid analgesic use (0.24). CONCLUSIONS: We found social and HIV-related variables associated with decreasing pain. We failed to show a positive association between analgesic use and decreasing pain. PMID- 21642848 TI - Boston type 1 keratoprosthesis combined with silicone oil for treatment of hypotony in prephthisical eyes. AB - PURPOSE: To present the outcomes of Boston type I keratoprosthesis (KPro) implantation in combination with pars plana vitrectomy (PPV) and silicone oil for the treatment of hypotony in prephthisical eyes. METHODS: Interventional case series. Thirteen eyes of 13 patients underwent Boston type I KPro implantation, pars plana vitrectomy, and silicone oil placement. Concurrent retinal detachment repair, membrane peel, or intraocular lens explantation were performed if necessary. Inclusion criteria for surgery were eyes with visual acuity worse than 20/400, previous failed penetrating keratoplasty, corneal opacification, visually significant or worsening hypotony, and visual acuity 20/200 or worse in the fellow eye. Outcome measures included Snellen best-corrected visual acuity, anatomic retinal attachment, and complications. RESULTS: At the final follow-up (mean, 24 months; range, 5-66 months), visual acuity was improved in 10 of 13 eyes (77%), stable in 2 of 13 eyes (15%), and decreased in 1 of 13 eyes (8%). All eyes had attached retina with no progression to phthisis bulbi. No intraoperative complications occurred. Postoperative complications included retroprosthetic membrane (7 of 13), KPro melt (1 of 13), KPro leak (1 of 13), KPro infection (1 of 13), vitreous hemorrhage (1 of 13), and retinal detachment (1 of 13). CONCLUSIONS: Boston type I KPro implantation in combination with pars plana vitrectomy and intraocular silicone oil fill can improve vision in most prephthisical eyes with hypotony. Structural findings can also improve. PMID- 21642849 TI - Cornea Society nomenclature for ocular surface rehabilitative procedures. AB - PURPOSE: In the past 20 years, there has been tremendous development in ocular surface rehabilitation and, through better understanding and improvements in analytic and culture techniques, a variety of new procedures have been developed. Differing techniques have been used internationally and often similar terminology is used when procedures could be considered to be quite different or vice versa. To communicate clearly and to compare techniques and outcomes, it was determined that an agreed international nomenclature was necessary. METHODS: A subcommittee was established by The Cornea Society. An initial steering group of international experts with special interest in ocular surface was assembled and established broad principles for the nomenclature based on a previous nomenclature. RESULTS: The nomenclature for procedures was based on several parameters, including a) anatomic source of tissue transplanted, for example conjunctival, keratolimbal or mucosal, b) whether the source was autologous or allogeneic (cadaveric or living related), and c) cell culture techniques. For example, an expanded cell culture of cadaveric limbal tissue was named ex-vivo cultivated cadaveric limbal allograft (EvC-LAL). CONCLUSIONS: Agreed nomenclature for transplantation procedures used in ocular surface rehabilitation has been developed, and use of this common terminology should help communication among those involved in this field. PMID- 21642850 TI - Conjunctival proliferation after a mild pepper spray injury in a young child. AB - PURPOSE: To report a case of conjunctival proliferation in a 2.5-year-old boy after initial evidence of a mild chemical injury after ocular exposure to pepper spray (oleoresin capsicum). METHODS: Case report with ophthalmologic and histologic findings. RESULTS: A child presented with mild conjunctival injection and chemosis without any corneal erosion after direct exposure to pepper spray. Three weeks later, a significant conjunctival proliferation was found at the limbus, which was refractory to treatment with topical corticosteroids. Finally, proliferative tissue was surgically excised without clinical recurrence during 2 months of follow-up. CONCLUSIONS: We hypothesize that the young age of the patient may have been an important factor for the severe conjunctival proliferation in comparison to a mainly uncomplicated course of pepper spray injuries in most adults. We recommend the use of topical antiinflammatory treatment even in apparently mild pepper spray injuries, especially in young children. PMID- 21642851 TI - Central corneal thickness in patients with systemic sclerosis: a controlled study. AB - PURPOSE: To compare the central corneal thickness (CCT) of patients with systemic sclerosis (SSc) and control subjects. METHODS: The study group comprised 37 consecutive patients with SSc, and the control group comprised 23 healthy individuals similar in age and sex. CCT value was measured by ultrasound pachymetry. RESULTS: In the SSc group, the mean CCT in the right eye was 534.9 +/ 33.5 MUm and 536.9 +/- 32.4 MUm in the left eye. In the control group, the mean CCT was 533.0 +/- 32.9 MUm in the right eye and 533.1 +/- 33.6 MUm in the left eye. The mean CCT was not significantly different in the SSc group compared with the control group for both the right (P = 0.83) and left (P = 0.67) eyes. CONCLUSIONS: CCT measurements do not significantly differ in patients with SSc compared with healthy control subjects. PMID- 21642853 TI - Large-bubble modification of the big-bubble technique. PMID- 21642854 TI - Cognitive-behavioral classifications of chronic pain in patients with multiple sclerosis. AB - The aim of this study was to replicate, in patients with multiple sclerosis (MS), the three-cluster cognitive-behavioral classification proposed by Turk and Rudy. Sixty-two patients attending a tertiary MS rehabilitation center completed the Pain Impact Rating questionnaire measuring activity interference, pain intensity, social support, and emotional distress. The General Health Questionnaire-28 and the Multiple Sclerosis Impact Scale-29 assessed disability and restriction in participation. Cluster analysis classified patients into three cognitive behavioral groups (40.4%, 'adaptive copers'; 36.5%, 'dysfunctional'; and 23.1%, 'interpersonally distressed'). Patients in groups with higher levels of activity interference, emotional distress due to pain, and lower perceived levels of social support had significantly higher levels of depression on the General Health Questionnaire-28 (P<0.003), and reported a greater impact on their physical and psychological functioning (P<0.001) on Multiple Sclerosis Impact Scale-29 subscales. Possible cut-points were identified to aid clinicians in classifying patients into clusters for individualized treatment. More research is needed to improve the understanding of pain and the potential use of cognitive behavioral clusters in patients with MS. These may be useful in the development of tailored early intervention, which may reduce pain-related disability and contribute to patient's overall well being. PMID- 21642855 TI - Diffusion tensor imaging findings are not strongly associated with postconcussional disorder 2 months following mild traumatic brain injury. AB - OBJECTIVE: To examine the relation between diffusion tensor imaging (DTI) of the corpus callosum and postconcussion symptom reporting following mild traumatic brain injury (MTBI). PARTICIPANTS: Sixty patients with MTBI and 34 patients with orthopedic/soft-tissue injuries (Trauma Controls) prospectively enrolled from consecutive admissions to a level 1 trauma center. PROCEDURE: Diffusion tensor imaging of the corpus callosum was undertaken using a Phillips 3T scanner at 6 to 8 weeks postinjury. Participants also completed a postconcussion symptom checklist. The MTBI group was divided into 2 subgroups based on the International Classification of Diseases, Tenth Revision symptom criteria for postconcussion disorder (PCD): PCD Present (n = 21), PCD Absent (n = 39). MAIN OUTCOME MEASURES: Measures of fractional anisotropy and mean diffusivity for the genu, body, and splenium of the corpus callosum. Participants also completed the British Columbia Post-Concussion Symptom Inventory. RESULTS: The MTBI group reported more postconcussion symptoms than the trauma controls. There were no significant differences between MTBI and trauma control groups on all DTI measures. In the MTBI sample, there were no significant differences on all DTI measures between those who did and did not meet the International Classification of Diseases, Tenth Revision research criteria for postconcussion disorder. CONCLUSIONS: These data do not support an association between white matter integrity in the corpus callosum and self-reported postconcussion syndrome 6 to 8 weeks post-MTBI. PMID- 21642856 TI - Perthes-like disease in Alagille syndrome. AB - We describe a unique case of a bilateral osteochondrosis of the femoral heads, similar to Perthes disease, in a boy affected by Alagille syndrome. This is a rare genetic syndrome, caused by vascular anomalies, and characterized by five main features: hepatic, cardiovascular, ophthalmological, skeletal malformations, and characteristic facial appearance. The most frequent skeletal finding is the 'butterfly vertebra'. We have followed the patient from the age of 5 years to the age of 20 years. We performed two bilateral valgus osteotomies when he was 10 years old to limit the progression of the deformity. We believe that the association of a bilateral osteochondrosis of the femoral heads with Alagille syndrome, a disease characterized by a vascular etiology, supports the hypothesis of angiogenic pathogenesis of Perthes disease. PMID- 21642857 TI - Personal best marathon time and longest training run, not anthropometry, predict performance in recreational 24-hour ultrarunners. AB - In recent studies, a relationship between both low body fat and low thicknesses of selected skinfolds has been demonstrated for running performance of distances from 100 m to the marathon but not in ultramarathon. We investigated the association of anthropometric and training characteristics with race performance in 63 male recreational ultrarunners in a 24-hour run using bi and multivariate analysis. The athletes achieved an average distance of 146.1 (43.1) km. In the bivariate analysis, body mass (r = -0.25), the sum of 9 skinfolds (r = -0.32), the sum of upper body skinfolds (r = -0.34), body fat percentage (r = -0.32), weekly kilometers ran (r = 0.31), longest training session before the 24-hour run (r = 0.56), and personal best marathon time (r = -0.58) were related to race performance. Stepwise multiple regression showed that both the longest training session before the 24-hour run (p = 0.0013) and the personal best marathon time (p = 0.0015) had the best correlation with race performance. Performance in these 24-hour runners may be predicted (r2 = 0.46) by the following equation: Performance in a 24-hour run, km) = 234.7 + 0.481 (longest training session before the 24-hour run, km) - 0.594 (personal best marathon time, minutes). For practical applications, training variables such as volume and intensity were associated with performance but not anthropometric variables. To achieve maximum kilometers in a 24-hour run, recreational ultrarunners should have a personal best marathon time of ~3 hours 20 minutes and complete a long training run of ~60 km before the race, whereas anthropometric characteristics such as low body fat or low skinfold thicknesses showed no association with performance. PMID- 21642858 TI - Heart rate responses and technical comparison between small- vs. large-sided games in elite professional soccer. AB - This study aims to examine the difference in heart rate (HR) responses and technical activities placed upon European elite players when exposed to 2-sided games differing in the number of players and playing area. Fifteen male soccer players from a Scottish Premier League team (26.3 +/- 4.85 years, 182.4 +/- 6.99 cm, 79.5 +/- 8.05 kg, VO2max of 54.36 +/- 5.45 ml.kg(-1).min(-1)) performed both small (3 vs. 3 plus goalkeepers) and large (9 vs. 9 plus goalkeepers) sided games each lasting for 3 * 5 minutes interspersed with 4-minute passive recovery during the 2009-2010 season. The HR responses and players' technical actions were recorded throughout all sided games. Results show that small-sided games (SSG) induced significantly (p < 0.05, large effect) higher HR responses as compared to large-sided games. Furthermore, during SSGs, players spent significantly longer time in the >85% maximal HR zone (p < 0.05, large effect) as compared to large sided games. Technical analysis revealed a large practical difference (effect size ranged from 1.5 to 21.2) between small- and large-sided games: less number of blocks, headers, interceptions, passes, and receives but more dribbles, shots, and tackles in SSG. Furthermore, SSG induced significantly lesser total ball contacts per game (p < 0.05, large effect) but significantly greater ball contacts per individual (p < 0.05, large effect) when compared to larger-sided games. The different technical requirements also enable coaches to carry out training games more suitable to specific playing positions such as SSG for midfielders (more dribbles, tackles, and ball contacts per player) strikers (more shots), and large-sided games for defender (more blocks, headers, and interceptions). PMID- 21642859 TI - Does lung adenocarcinoma subtype predict patient survival?: A clinicopathologic study based on the new International Association for the Study of Lung Cancer/American Thoracic Society/European Respiratory Society international multidisciplinary lung adenocarcinoma classification. AB - INTRODUCTION: Lung adenocarcinoma is a heterogeneous group of tumors with a highly variable prognosis, not well predicted by the current pathologic classification system. The 2004 World Health Organization classification results in virtually all tumors encountered in clinical practice being allocated to the adenocarcinoma of mixed subtype category. A new classification developed by an international multidisciplinary expert panel sponsored by the International Association for the Study of Lung Cancer, American Thoracic Society, and European Respiratory Society, is based on histomorphologic subtype and has recently been validated in a North American series of 514 stage I lung adenocarcinomas. We investigated the relationship between the new classification and patient survival in a series of Australian patients with stages I, II, and III lung adenocarcinoma. METHODS: We identified 210 patients from a surgical database who underwent resection of lung adenocarcinoma from 1996 to 2009. Two pathologists, blinded to patient outcome, independently performed histopathologic subtyping according to the new classification. Kaplan-Meier curves were used to calculate 5 year survival for each separate histopathologic subtype/variant. Univariate and multivariate analyses were undertaken to control for validated prognostic factors. RESULTS: We confirmed that the new subtypes of adenocarcinoma in situ, minimally invasive adenocarcinoma and lepidic-predominant adenocarcinoma had a 5 year survival approaching 100%, whereas micropapillary-predominant and solid with mucin-predominant adenocarcinomas were associated with particularly poor survival. Papillary-predominant and acinar-predominant adenocarcinomas had an intermediate prognosis. This effect persisted after controlling for stage. CONCLUSIONS: Classification of lung adenocarcinoma according to the new International Association for the Study of Lung Cancer/American Thoracic Society/European Respiratory Society classification correlated with 5-year survival. These relationships persisted after controlling for known prognostic patient and tumor characteristics. The new classification has advantages not only for individual patient care but also for better selection and stratification for clinical trials and molecular studies. PMID- 21642860 TI - The Association of Intratumoral Germinal Centers with early-stage non-small cell lung cancer. AB - INTRODUCTION: Lung cancers can display immune cell infiltration although the role of an adaptive immune response in disease pathogenesis is unknown. To investigate the possibility of a functional humoral response to the tumor, we surveyed histologic sections from non-small cell lung cancer (NSCLC) tumors for germinal centers (GCs) and assessed whether there was an association between the presence of GCs and tumor stage. METHODS: Tumor sections from 91 patients with all stages of NSCLC were examined by a pathologist blinded to clinical data. GCs were identified by hematoxylin and eosin staining patterns and confirmed by immunohistochemical staining for B-cell markers, BCL-6 and CD21. The distribution of GCs within the tumor or the tumor margin was recorded. Statistical analysis was performed to evaluate the association between stage and presence of GCs. RESULTS: Thirty-five percent of all tumors evaluated contained GCs, and sections evaluated by immunohistochemistry showed positive staining for both B-cell markers. GCs were seen both within the tumor and the tumor margin, consistent with an immune response to antigen stimulation. Patients with stage I NSCLC had a higher prevalence of intratumoral GCs than patients with stages II to IV (Cochran Armitage Trend Test p = 0.02011). There was no association of stage with GCs in the tumor margin. CONCLUSIONS: Intratumoral GCs are associated with early-stage NSCLC. Further characterization of intratumoral GCs may lead to new diagnostic and therapeutic strategies based on manipulation of the adaptive immune response. PMID- 21642861 TI - Histone deacetylase inhibitors sensitize human non-small cell lung cancer cells to ionizing radiation through acetyl p53-mediated c-myc down-regulation. AB - INTRODUCTION: Histone deacetylase inhibitors (HDACIs) induce growth arrest and apoptosis in cancer cells. In addition to their intrinsic anticancer properties, HDACIs modulate cellular responses to ionizing radiation (IR). We examined the molecular mechanism(s) associated with the radiosensitizing effects of HDACIs in human lung cancer cells. METHODS: Lung cancer cells were pretreated with the appropriate concentrations of suberoylanilide hydroxamic acid or trichostatin A. After 2 hours, cells were irradiated with various doses of gamma-IR, and then we performed 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, fluorescence-activated cell sorting analysis, clonogenic assay, and Western blotting to detect cell viability or apoptosis and changes of specific proteins expression levels. RESULTS: In this study, we showed that HDACIs (including suberoylanilide hydroxamic acid and trichostatin A) and IR synergistically trigger cell death in human non-small cell lung cancer cells. Cell viability and clonogenic survival were markedly decreased in cultures cotreated with HDACIs and IR. Interestingly, p53 acetylation at lysine 382 was significantly increased, and c-myc expression simultaneously down-regulated in cotreated cells. Radiosensitization by HDACIs was inhibited on transfection with small interfering RNA against p53 and c-myc overexpression, supporting the involvement of p53 and c myc in this process. Furthermore, c-myc down-regulation and apoptotic cell death coinduced by IR and HDACI were suppressed in cells transfected with mutant K382R p53 and C135Y p53 displaying loss of acetylation at lysine 382 and DNA-binding activity, respectively. CONCLUSIONS: Our results collectively demonstrate that the degree of radiosensitization by HDACIs is influenced by acetyl p53-mediated c myc down-regulation. PMID- 21642862 TI - Impact of induction chemotherapy on estimated risk of radiation pneumonitis in small cell lung cancer. AB - INTRODUCTION: Induction chemotherapy in patients with bulky small cell lung cancer (SCLC) could lead to decreased tumor burden, smaller radiation fields, and less pulmonary toxicity. This study compared radiation therapy (RT) plans based on pre- and postchemotherapy computed tomography (CT) scans of patients with SCLC to estimate the reduced risk of radiation pneumonitis (RP) after receiving chemotherapy. METHODS: Between 2003 and 2009, 23 patients with stage IV SCLC were treated with chemotherapy alone (no surgery or RT) and had computed tomography scans pre- and post two cycles of platin-based chemotherapy. Simulated RT plans were created as if to deliver 45 Gy to the thoracic disease. The percent of lung receiving >=20 Gy (V20), mean lung dose, and normal tissue complication probability (NTCP) was evaluated in patients who had a partial response (>=30% volumetric reduction) in gross tumor volume. RESULTS: One (4.3%) patient had a complete response, 18 (78.3%) had a partial response, and four (17.4%) had stable disease. Among 18 responders, the absolute decrease in V20 was 7.4% (p < 0.01), in mean lung dose was 3.3 Gy (p < 0.01), and in NTCP was 5.5% (p < 0.01). Patients with a prechemotherapy V20 >=35% versus V20 less than 35% had an average absolute reduction in NTCP of 10% versus 2% (p < 0.01). CONCLUSION: Patients with limited stage SCLC with a V20 >=35% may benefit from induction chemotherapy as there is an estimated reduction of RP of 10%. This reduction in risk of RP after induction chemotherapy should be weighed against risks and benefits of delaying upfront RT. PMID- 21642863 TI - The value of autofluorescence bronchoscopy combined with white light bronchoscopy compared with white light alone in the diagnosis of intraepithelial neoplasia and invasive lung cancer: a meta-analysis. AB - OBJECTIVE: To compare the accuracy of autofluorescence bronchoscopy (AFB) combined with white light bronchoscopy (WLB) versus WLB alone in the diagnosis of lung cancer. METHODS: The Ovid, PubMed, and Google Scholar databases from January 1990 to October 2010 were searched. Two reviewers independently assessed the quality of the trials and extracted data. The relative risk for sensitivity and specificity on a per-lesion basis of AFB + WLB versus WLB alone to detect intraepithelial neoplasia and invasive cancer were pooled by Review Manager. RESULTS: Twenty-one studies involving 3266 patients were ultimately analyzed. The pool relative sensitivity on a per-lesion basis of AFB + WLB versus WLB alone to detect intraepithelial neoplasia and invasive cancer was 2.04 (95% confidence interval [CI] 1.72-2.42) and 1.15 (95% CI 1.05-1.26), respectively. The pool relative specificity on a per-lesion basis of AFB + WLB versus WLB alone was 0.65 (95% CI 0.59-0.73). CONCLUSIONS: Although the specificity of AFB + WLB is lower than WLB alone, AFB + WLB seems to significantly improve the sensitivity to detect intraepithelial neoplasia. However, this advantage over WLB alone seems much less in detecting invasive lung cancer. PMID- 21642864 TI - Does anatomical segmentectomy allow an adequate lymph node staging for cT1a non small cell lung cancer? AB - INTRODUCTION: Anatomical segmentectomy is again under evaluation for the cure of T1a N0 non-small cell lung cancer and carcinoid tumors. Whether anatomical segmentectomy does permit or not, an adequate resection of nodal stations for staging or cure is still pending. METHODS: A case-matched study was ruled on patients with peripheral cT1a N0 M0 tumors that underwent anatomical segmentectomy or lobectomy. Dissection of lymph node stations 4, 5, 6, and 7 was identical in anatomical segmentectomy and lobectomy; stations 10, 11, 12, and 13 were also dissected carefully during anatomical segmentectomy. RESULTS: We individually matched 46 (69% men) anatomical segmentectomy with 46 (71% men) lobectomy for age, anatomical segment, and size of the tumor. The median (interquartile range) size of the resected lesions was 1.7 cm (1.35-1.95 cm) in anatomical segmentectomy and 1.6 cm (1.3-1.9 cm) (p = 0.96) in lobectomy. The anatomical segmentectomy and lobectomy resection margins were free of cancer. The median number (interquartile range) of total dissected lymph nodes was 12 (8-5 14) in anatomical segmentectomy compared with 13 (12-14.5) in lobectomy (p = 0.68), with a number of N1 nodes being 6 (4-7.5) and 7 (4.5-9.5) (p = 0.43), respectively, and N2 nodes 5.5 (4-7.7) and 5 (4-6.5) (p = 0.88). Only 1 patient of 46 (2%) anatomical segmentectomy was N1, whereas in lobectomy, 4% had N1 (2 patients). Freedom from recurrence at 36 months was 100% for anatomical segmentectomy and 93.5% for lobectomy (p = 0.33). CONCLUSIONS: Anatomical segmentectomy for cT1a tumors compared with lobectomy procures an adequate number of N1 and N2 nodes for pathological examination. Cancer-specific survival was equivalent at 36 months. PMID- 21642865 TI - Anaplastic lymphoma kinase translocation: a predictive biomarker of pemetrexed in patients with non-small cell lung cancer. AB - INTRODUCTION: This study compared the efficacy of pemetrexed in patients with anaplastic lymphoma kinase (ALK)-positive versus ALK-negative (epidermal growth factor receptor [EGFR] mutant or wild type [WT] for both ALK and EGFR) non-small cell lung cancer (NSCLC). METHODS: Patients with advanced NSCLC who received second-line pemetrexed and beyond between March 2007 and April 2010 were screened for EGFR mutations and ALK rearrangements at Seoul National University Hospital. The clinical and in vitro efficacy of pemetrexed was evaluated for each genotypic group. RESULTS: Ninety-five NSCLC patients were genotyped as follows: 43 (45%) EGFR mutation, 15 (16%) ALK translocation, and 37 (39%) WT. The overall response rate was superior in ALK-translocated patients compared with EGFR mutant or WT patients (46.7 versus 4.7 versus 16.2%, p = 0.001). ALK-positive patients showed longer time to progression than EGFR mutant or WT patients (9.2 versus 1.4 versus 2.9 months, p = 0.001). ALK positivity alone was a significant predictor for overall response rate (hazard ratio [HR] = 0.07, 95% confidence interval [CI]: 0.01-0.32; p = 0.001) and time to progression (HR = 0.44, 95% CI: 0.24-0.80; p = 0.007). ALK positivity remained independently significant regardless of treatment line (HR = 0.43, 95% CI: 0.24-0.77; p = 0.005). Thymidylate synthase mRNA levels in ALK-positive cells were significantly lower compared with control cells (p < 0.05). CONCLUSION: Pemetrexed is an effective treatment in patients with ALK positive NSCLC. ALK positivity was independently predictive of pemetrexed efficacy in NSCLC patients. PMID- 21642866 TI - The zinc ionophore PCI-5002 radiosensitizes non-small cell lung cancer cells by enhancing autophagic cell death. AB - INTRODUCTION: A major focus of cancer research is to identify compounds that sensitize resistant cancer cells to radiation treatment. Lung cancer cells, in particular, have high rates of radioresistance that lead to treatment failure. We have previously shown that the autophagy induced in the context of decreased apoptosis confers radiosensitivity to prostate and lung cancer cells. Zinc supplementation has antiapoptotic effects in cell culture. In addition, the accumulation of zinc in response to oxidative stress has been associated with increased autophagy in astrocyte and breast cancer cells. METHODS: In this study, we hypothesized that the zinc ionophore PCI-5002 radiosensitizes lung cancer cells by inducing autophagic cell death. To test this hypothesis, we used a combination of in vitro and in vivo approaches, including clonogenic assays to test for radiosensitivity, biochemical analyses of apoptosis and autophagy, and a xenograft mouse model of tumor growth. RESULTS: We found that PCI-5002 reduced clonogenic survival in treated cells compared with untreated cells (0.03% versus 0.1% surviving fraction, p < 0.001). The increased radiosensitive fraction of PCI 5002-treated cells was accompanied by increased autophagy. PCI-5002 treatment also reduced caspase-3 cleavage. In an irradiated xenograft mouse model, the tumor growth of irradiated, PCI-5002-treated mice was slower than untreated, irradiated mice (25 days versus 22 days to reach a 1.0 cm tumor size). CONCLUSIONS: PCI-5002 treatment sensitizes lung cancer cells to radiation, both in vitro and in vivo. This data suggest that PCI-5002 could potentially treat radioresistant/locally advanced lung cancer by amplifying the effects of radiotherapy. PMID- 21642867 TI - Tumor cell repopulation between cycles of chemotherapy is inhibited by regulatory T-cell depletion in a murine mesothelioma model. AB - INTRODUCTION: Malignant pleural mesothelioma is a highly aggressive cancer with poor prognosis. We have previously demonstrated that regulatory T cells (Treg) depletion can impact tumor microenvironment when combined with chemotherapy. The aim of this study is to analyze the impact of Treg depletion on tumor cell repopulation during cycles of chemotherapy in a murine mesothelioma model. METHODS: Tumor-bearing mice were treated with chemotherapy once weekly to mimic clinical settings and with PC61 to cause Treg depletion after each cycle of chemotherapy. Tumor cell repopulation was evaluated by BrdU labeling index with immunohistochemistry and flow cytometry, and Ki67 gene expression was determined by real-time reverse-transcribed polymerase chain reaction. The proportion of CD4+ CD25+Foxp3+ Tregs, CD4+, and CD8+ T cells in the tumor, spleen, draining lymph node, and peripheral blood from tumor-bearing mice was determined by using flow cytometry, and gene expression of activated T-cell-related cytokines was quantified by enzyme-linked immunosorbent assay and reverse-transcribed polymerase chain reaction. RESULTS: Tumor growth delay was achieved by cisplatin followed by PC61 or cyclophosphamide. The BrdU labeling index indicated that tumor cell repopulation between cycles of cisplatin was significantly inhibited by PC61. The CD4+CD25+Foxp3+ Tregs in tumor and lymphoid organs were almost completely depleted, whereas the CD4+ or CD8+ T cells did not change. PC61 after chemotherapy resulted in an increase of gene expression of interferon-gamma, granzyme B, perforin, and IP-10, thus leading to tumor cell lysis in cytotoxic lymphocyte assay. Nevertheless, cell killing induced by cyclophosphamide combined with cisplatin was due to cytotoxicity rather than specific immune response. CONCLUSION: Treg depletion between cycles of chemotherapy could improve the outcome of mesothelioma. Nevertheless, this effect seems limited, and more effective approaches need to be developed. PMID- 21642868 TI - Nutrition during trimodality treatment in stage III non-small cell lung cancer: not only important for underweight patients. AB - INTRODUCTION: Trimodality treatment for stage III non-small cell lung cancer (NSCLC), consisting of chemoradiotherapy followed by surgery, is associated with treatment-related toxicity, malnutrition, and postoperative complications. The aim of this retrospective study was to investigate the predictive value of nutritional parameters on postoperative morbidity, mortality, and survival. METHODS: Patients with stage III NSCLC undergoing concurrent chemoradiotherapy followed by surgery in one center between 2003 and 2009 were included. Age, sex, forced expiratory volume in 1 second, body mass index, weight change, and surgical and pathological factors were recorded and related to the occurrence of postoperative complications/mortality, overall survival (OS), and progression free survival. RESULTS: Of 51 study patients, 17 (33%) had overweight (body mass index >= 25) at start of treatment and 20 patients (39%) were malnourished at hospital admission for surgery. Postoperative complications occurred in 25 patients (49%), 6 had major complications, and 2 died within 90 days after surgery, but no significant predictive factors were found. Overall, weight loss >=5% during induction period was associated with shorter OS (p = 0.03), but especially overweight patients experiencing weight loss >=5% during induction period (n = 7) had shorter OS (hazard ratio 4.63, p = 0.005; log-rank p = 0.04) and progression-free survival (hazard ratio 6.03, p = 0.007). CONCLUSIONS: This study indicates that malnutrition especially in overweight patients negatively influences survival outcomes of trimodality treatment for stage III NSCLC. PMID- 21642869 TI - A 10-year single-center experience on 708 lung metastasectomies: the evidence of the "international registry of lung metastases". AB - INTRODUCTION: The International Registry of Lung Metastases defined a new staging system based on identified prognostic factors for long-term survival after metastasectomy. The aim of our study was to confirm the validity of the International Registry of Lung Metastases classification system in patients who underwent curative lung metastasectomy in a single center. METHODS: We retrospectively reviewed 575 patients who underwent 708 lung metastasectomies from January 1998 to October 2008. Complete curative pulmonary resections were performed in 490 cases (85%). Three hundred seventy-two patients developed lung metastases from epithelial tumors, 80 from sarcomas, 27 from melanomas, and 11 from germ cell tumors. The mean disease-free interval (DFI) was 46.6 months. Open surgical resection was performed in 479 patients. One hundred eighty-five patients had a single-lung metastasis. Lymph node dissection was performed in 353 cases. RESULTS: After a mean follow-up of 34 months, 247 patients (43%) had died. Multivariate analysis disclosed that completeness of resection (p < 0.0001), patients with germ cell tumors (p = 0.04), and DFI >=36 months (p = 0.01) were also associated with a better prognosis. The actuarial survival after complete metastasectomy was 74% at 2 years and 46% at 5 years. CONCLUSIONS: We confirmed completeness of surgery, histology, and DFI >=36 months as independent prognostic factors. Number of metastases, presence of lymph node metastases, surgical approach, and number of metastasectomies did not statistically influence long term survival. PMID- 21642870 TI - Differential effect of polymorphisms of CMPK1 and RRM1 on survival in advanced non-small cell lung cancer patients treated with gemcitabine or taxane/cisplatinum. AB - INTRODUCTION: To determine whether genetic variations in CMPK1 or RRM1, which impact the pharmacodynamics of gemcitabine, differentially affect the outcomes of non-small cell lung cancer (NSCLC) patients treated with gemcitabine or taxane/cisplatinum. METHODS: We conducted retrospective study evaluating the associations between overall survival in 298 NSCLC patients at stages IIIA/IIIB (140) and IV (158), treated with gemcitabine (139) or taxane (159)/cisplatinum and 14 tagging single-nucleotide polymorphisms (tSNPs): 4 in CMPK1 and 10 in RRM1. RESULTS: The wild-type genotypes of CMPK1 IVS1+1057 and IVS1-928 were associated with shorter overall survival in patients treated with the gemcitabine/cisplatinum (adjusted hazards ratio = 1.97 and 1.89; Cox pBonferroni = 0.008 and 0.020), whereas this effect was not observed in patients treated with taxane/cisplatinum. No associations were observed for the other 2 CMPK1 or 10 RRM1 tSNPs. Analysis of the interaction between the CMPK1 and RRM1 genes showed that the survival of patients with CMPK1 IVS1+1057 CC and RRM1 IVS1-2374 TT, IVS7+25 AA, IVS7-425 AA, or IVS8+287 TT was significantly shorter when they were treated with the gemcitabine/cisplatinum (adjusted hazards ratio = 3.00, 2.89, 3.14, and 3.00; Cox pBonferroni = 0.007, 0.012, 0.006, and 0.007). However, these effects were not observed in patients treated with taxane/cisplatinum. CONCLUSIONS: These findings suggest that polymorphisms of CMPK1 and their combination with those of RRM1 are helpful in identifying patients who will benefit less from a gemcitabine/cisplatinum as the first-line regimen. PMID- 21642871 TI - Postoperative radiotherapy for completely resected stage II or III thymoma. AB - INTRODUCTION: We assessed the efficacy of adjuvant radiotherapy (RT) in patients with stage II or III thymoma and evaluated the prognostic factors after the treatment. PATIENTS AND METHODS: The medical records of 76 patients with completely resected stage II (n = 65) or III (n = 11) thymoma treated at a single institution within the period from November 1988 to February 2009 were retrospectively reviewed. Seventeen patients were treated with surgery only (group A), and 59 patients received adjuvant RT after surgery (group B). The median radiation dose was 50 Gy (range: 43.2-66 Gy). The prognostic factors, such as age, myasthenia gravis symptom, tumor size, staging, adjuvant RT, and radiation dose, were analyzed. The median follow-up time was 58.5 months (range: 6-231 months). RESULTS: The 5- to 10-year overall survival and disease-free survival (DFS) rates were 95.3% and 83.8% and 91.5% and 82.5%, respectively. The 5- and 10-year DFS (80% and 70% and 97.8% and 92.7% in groups A and B, respectively; p = 0.043), and the median time to recurrence (37.4 and 50.6 months in groups A and B, respectively) was statistically different between groups A and B. CONCLUSION: Masaoka staging and adjuvant RT were related to DFS in completely resected stage II or III thymoma. Based on this study, adjuvant RT will be beneficial in this clinical setting. PMID- 21642872 TI - Combined serum mesothelin and plasma osteopontin measurements in malignant pleural mesothelioma. AB - INTRODUCTION: Malignant pleural mesothelioma (MPM) is a lethal tumor related to asbestos exposure. At present, the only instruments for screening and diagnosis are based on radiological tests, posing evident economic and radio-protectionist problems. Some authors are evaluating biological indicators, such as plasma osteopontin (pOPN) and serum soluble mesothelin-related peptides (SMRP). This study aimed to evaluate whether a combination of these two markers could increase sensitivity and specificity in diagnosis of epithelioid MPM. METHODS: We enrolled 93 healthy subjects, 111 individuals with benign respiratory disease (BRD), and 31 patients with MPM, histologically and/or cytologically confirmed. SMRP and pOPN levels were determined using commercially available enzyme-linked immunosorbent assay kits. Though a logistic regression analysis, SMRP and pOPN were combined and translated into a new index, called "combined risk index." RESULTS: Differences in both SMRP and pOPN mean values between epithelial MPM patients and healthy subjects or BRD patients were statistically significant (p < 0.0001), whereas there was no difference in SMRP and pOPN mean values between healthy subjects and BRD patients. The performance in MPM diagnosis resulted improved by the combination of the two markers. The results of our study should be confirmed by a larger scale and, possibly, a multicenter study, which could better take into consideration the influence of some possible confounding factors such as glomerular filtration rate and other blood parameters. CONCLUSIONS: We combined SMRP and pOPN dosages to increase diagnostic accuracy. This study showed for the first time that combined SMRP and pOPN measurements can increase both sensitivity and specificity in terms of combined risk index. PMID- 21642873 TI - Transesophageal ultrasound-guided fine-needle aspiration for the mediastinal restaging of non-small cell lung cancer. AB - INTRODUCTION: Selected patients with stage III (N2/N3) non-small cell lung cancer (NSCLC) who are downstaged to N0 by chemoradiation therapy might benefit from subsequent surgical resection of the tumor. How mediastinal lymph nodes can be best reevaluated is subject of debate. Transesophageal ultrasound-guided fine needle aspiration (EUS-FNA) is a minimally invasive technique to sample mediastinal nodes. We assessed sensitivity and false-negative rate of EUS-FNA for the mediastinal restaging of patients with stage III NSCLC. METHODS: Fifty-eight consecutive patients with stage III NSCLC and tissue-proven lymph node metastases N2/N3) who underwent EUS-FNA for restaging purposes after chemoradiation therapy were retrospectively analyzed. Surgical-pathological staging was used as the reference standard for nodal metastases. RESULTS: EUS-FNA found persistent nodal metastases (N2/N3) in 15 patients (26%). Of the 43 patients without persistent mediastinal metastases at EUS, 33 patients subsequently underwent surgical verification of the mediastinal nodes in whom persistent metastases (yN2/N3) were found in 19 patients (58%), and loco-regional downstaging (yN0) was achieved in the other 14 (42%). The prevalence of persistent nodal metastases in the 48 patients who could be analyzed was 71%. Sensitivity and the false-negative rate of EUS-FNA for mediastinal restaging were 44 and 58%, respectively. DISCUSSION: For mediastinal restaging of stage III NSCLC, EUS-FNA is a minimally invasive and safe method to confirm persistent nodal metastases, but this technique has a low negative predictive value and is therefore not useful for the exclusion of mediastinal metastases. Surgical restaging is indicated in the absence of mediastinal metastases at EUS-FNA. PMID- 21642874 TI - Diagnostic performance of integrated positron emission tomography/computed tomography for mediastinal lymph node staging in non-small cell lung cancer: a bivariate systematic review and meta-analysis. AB - INTRODUCTION: Accurate clinical staging of mediastinal lymph nodes (MLNs) of patients with non-small cell lung cancer (NSCLC) is important in determining therapeutic options and prognoses. Integrated positron emission tomography and computed tomography (PET/CT) scanning is becoming widely used for MLN staging in patients with NSCLC. We performed a bivariate meta-analysis to determine the pooled sensitivity (SEN) and specificity (SPE) of this imaging modality. METHODS: The PubMed/MEDLINE, Embase, and SpringerLink databases were searched for articles related to PET/CT for MLN staging in patients with NSCLC. SEN and SPE were calculated for every study. Hierarchical summary receiver operating characteristic curves were used to summarize overall test performance and assess study quality. Potential between-study heterogeneity was explored by subgroup analyses. RESULTS: Fourteen of 330 initially identified reports were included in the meta-analysis. When we did not consider the unit of analysis, the pooled weighted SEN and SPE were 0.73 (95% confidence interval [CI]: 0.65-0.79) and 0.92 (95% CI: 0.88-0.94), respectively. In the patient-based data analysis, the pooled weighted SEN was 0.76 (95% CI: 0.65-0.84) and the pooled weighted SPE was 0.88 (95% CI: 0.82-0.92). In the MLN-based data analysis, the pooled SEN was 0.68 (95% CI: 0.56-0.78) and the pooled SPE was 0.95 (95% CI: 0.91-0.97). CONCLUSIONS: Integrated PET/CT is a relatively accurate noninvasive imaging technique, with excellent specificity for MLN staging in patients with NSCLC. Nevertheless, current evidence suggests that we should not depend on the results of PET/CT completely for MLN staging in patients with NSCLC. PMID- 21642875 TI - Evaluation of a VHA collaborative to improve follow-up after a positive colorectal cancer screening test. AB - BACKGROUND: In 2005, the Veterans Health Administration initiated a yearlong Colorectal Cancer Care Collaborative (C4) to improve timely follow-up after positive fecal occult blood tests. METHODS: Twenty-one facilities formed local quality improvement (QI) teams. Teams received QI training, created process flow maps, implemented process changes, and shared learning through 2 face-to-face meetings, conference calls, and a discussion board. We evaluated pre-post change in the timeliness of follow-up among C4 facilities and 3 control facilities. Outcome measures included the proportion of patients receiving a follow-up colonoscopy within 1 year, the proportion receiving 60-day follow-up (the focus of C4 teams), and average days to colonoscopy. Survey data from C4 team members was analyzed to identify predictors of facility-level improvement. RESULTS: Both C4 and control facilities improved on 1-year follow-up (10% and 9% increases, respectively, both P's<0.001). There was a statistically significant increase in the proportion receiving 60-day follow-up among C4 facilities (27% pre-C4 vs. 39% post-C4, P=0.008) but a nonsignificant decrease among control facilities (45% pre C4 vs. 29% post-C4, P=0.14). Average days to colonoscopy decreased significantly among C4 facilities (129 pre-C4 vs. 103 post-C4, P=0.004) but increased significantly among control facilities (81 pre-C4 vs. 103 post-C4, P=0.04). Teams with the most improvement established clear roles/goals, had previous QI training, made more use of QI tools, and incorporated primary care education into their improvement work. CONCLUSIONS: A Veterans Health Administration improvement collaborative modestly decreased time to colonoscopy after a positive colorectal cancer screening test but significant room for improvement remains and benefits of participation were not realized by all facilities. PMID- 21642876 TI - Can we rely on patients' reports of adverse events? AB - BACKGROUND: Evidence suggests that patients can report a variety of adverse events (AEs) not captured by traditional methods such as a chart review. Little is known, however, about whether patient reports are useful for measuring patient safety. OBJECTIVES: To examine the degree to which physician reviewers agreed that patient reports of "negative effects" constituted AEs, and to identify questionnaire items that affected reviewers' judgments. METHODS: We surveyed patients discharged from Massachusetts hospitals in 2003 to elicit information about negative effects associated with hospitalization. Physician reviewers judged whether patient-reported negative effects represented AEs, and classified the severity of the event. Likelihood ratios were calculated to assess whether patient responses to questionnaire items affected reviewers' judgments. RESULTS: Of the 2582 patients surveyed, 753 patients reported 1170 negative effects, and 71.2% of these effects were classified as AEs by physician reviewers. Negative effects most likely to be classified as AEs involved newly prescribed medications and changes to previously prescribed medications. Additional information elicited from follow-up survey questions modestly affected reviewers' classification of serious AEs. Negative effects reported by women, younger patients, those reporting better health status, and those not admitted through the emergency department were more likely to be classified as AEs. CONCLUSIONS: Many patients were able to identify care-related AEs. Patient responses to questions about the sequelae of the events provided limited additional information for physicians to use in gauging the presence and severity of the event. Patient reports complement other incident-detection methods by providing information that is credible and unavailable from other sources. PMID- 21642877 TI - Shifts in surgical revascularization and valve procedures among Medicare beneficiaries. AB - BACKGROUND: The rapid development of percutaneous coronary intervention (PCI) has reduced the demand for coronary artery bypass grafting (CABG) surgery. We examined whether the previously documented declining demand for CABG and expanding PCI use are associated with changes in performance of valve (aortic or mitral) replacement or repair surgery. METHODS: We used Medicare Part A administrative data to identify beneficiaries who received PCI, CABG, valve procedures between 1991 and 2005. We used multivariable models to investigate the relationship between CABG and PCI rates and rates of aortic and mitral valve surgery at the hospital referral region level while adjusting for demand and supply factors including heart disease rates. RESULTS: Hospital referral region level rates of valve replacement and repair increased by 0.36 and 0.17 per 1000. A 1 per 1000 change in the isolated CABG rates was positively associated with a 0.0255 and 0.0013 per 1000 change in the concomitant (simultaneous CABG) valve replacement and repair rates. A 1 per 1000 change in the PCI rate was positively associated with 0.0105 and a 0.0051 per 1000 change in the concomitant valve replacement and repair rate. A 1 per 1000 change in the isolated CABG rates was positively associated with a 0.0151 per thousand isolated valve replacement and negatively associated with a 0.0013 per 1000 repair rate. A 1 per 1000 change in the PCI rate was positively associated with a 0.0081 and a 0.0031 per 1000 change in the isolated valve replacement and repair rate. CONCLUSIONS: A modest portion of the change in valve replacement and repair use over the period is associated with shifts in revascularization. Future study needs to consider the effects of these trends on patient outcomes and resource use. PMID- 21642878 TI - Association of MMP8 gene variation with an increased risk of malignant melanoma. AB - Matrix metalloproteinases (MMPs) are implicated in the development of cancers including malignant melanoma (MM) and breast cancer. We tested the possible association of MMP1 and MMP8 gene variation with these two types of cancer. We genotyped 300 unselected patients with MM, 300 consecutive breast cancer cases, 300 controls for melanoma, and 300 controls for breast cancer (age-matched and sex-matched healthy adults with negative cancer family histories). Our study showed that the MMP8 gene rs11225395 polymorphism was associated with the risk of developing MM (odds ratio: 1.69; 95% confidence interval: 1.02-2.80; P=0.040) for the A/A genotype and 1.49 (95% confidence interval: 1.03-2.17; P=0.035) for the A/G genotype compared with the G/G genotype. The A allele was over-represented among MM cases compared with controls (odds ratio=1.54; P=0.017). In-vitro assays showed that the A allele had a higher promoter activity than the G allele in melanoma cells. No association was detected between this variant and breast cancer susceptibility. We found no strong association between MMP1 variation and the risk of MM or breast cancer. The finding of this study indicates an influence of MMP8 gene variation on melanoma susceptibility. PMID- 21642879 TI - Sleep deprivation differentially affects dopamine receptor subtypes in mouse striatum. AB - The effects of sleep deprivation on dopaminergic systems remain elusive, in part due to the lack of selective ligands for dopamine receptor subtypes. We examined D1, D2, and D3 receptor density in the mouse brain after sleep deprivation by receptor autoradiography using [H]SCH 23390 for D1R, [H]raclopride for D2R, and [H]WC-10 for D3R (a novel D3R-selective compound developed in our laboratory, not previously reported in mouse). Sleep-deprived mice showed a significant decrease in D1R, no change in D2R, and a significant increase in D3R binding in striatum. This pattern of dopamine receptor changes was not seen in mice subjected to restraint stress, suggesting specificity to sleep. These data provide evidence that brain dopaminergic circuits are remodeled after sleep deprivation. PMID- 21642880 TI - Functional asymmetry in primary auditory cortex for processing musical sounds: temporal pattern analysis of fMRI time series. AB - Hemispheric differences in the temporal processing of musical sounds within the primary auditory cortex were investigated using functional magnetic resonance imaging (fMRI) time series analysis on a 3.0 T system in right-handed individuals who had no formal training in music. The two hemispheres exhibited a clear-cut asymmetry in the time pattern of fMRI signals. A large transient signal component was observed in the left primary auditory cortex immediately after the onset of musical sounds, while only sustained activation, without an initial transient component, was seen in the right primary auditory cortex. The observed difference was believed to reflect differential segmentation in primary auditory cortical sound processing. Although the left primary auditory cortex processed the entire 30-s musical sound stimulus as a single event, the right primary auditory cortex had low-level processing of sounds with multiple segmentations of shorter time scales. The study indicated that musical sounds are processed as 'sounds with contents', similar to how language is processed in the left primary auditory cortex. PMID- 21642881 TI - Functional study of endothelin B receptors in satellite glial cells in trigeminal ganglia. AB - There is immunohistochemical evidence for endothelin (ET) receptors in satellite glial cells in sensory ganglia, but there is no information on the function of these receptors. We used calcium imaging to study this question in isolated mouse trigeminal ganglia and found that satellite glial cells are highly sensitive to ET-1, with threshold at 0.05 nM. Responses displayed strong desensitization at ET 1 concentrations of more than 1 nM. A large component of the response persisted when Ca was deleted from the external medium, consistent with Ca release from internal stores. The use of receptor selective agents showed that the responses were mediated by ETB receptors. We conclude that satellite glial cells display endothelin receptors, which may participate in neuron-glia communications in the trigeminal ganglia. PMID- 21642882 TI - Diagnostic value of 99mTc-ubiquicidin scintigraphy for osteomyelitis and comparisons with 99mTc-methylene diphosphonate scintigraphy and magnetic resonance imaging. AB - OBJECTIVE: The discrimination of bacterial infections from sterile inflammatory processes is of great importance in the management of inflammation. Currently available techniques cannot decisively address this issue. In this respect, antimicrobial peptide Tc-ubiquicidin (UBI) 29-41 scans have been showing interesting results. The aim of this study was to determine the accuracy of Tc UBI scan in the detection of osteomyelitis and to compare it with Tc-methylene diphosphonate scan and magnetic resonance imaging (MRI). METHODS: Twenty patients (mean age=48.90 years) with suspected osteomyelitis were included in this study. After evaluation of each patient through history taking, physical examination, appropriate laboratory tests, and other processes including bone probing, wound culture, and plain film radiography, MRIs, Tc-UBI scans, and Tc-methylene diphosphonate scans were performed. For quantitative analysis, the mean count of abnormal-to-normal (A/N) region was calculated for images acquired at 15, 30, 45, 60, 120, and 240 min to obtain the most favorable time for imaging. RESULTS: In total, osteomyelitis was detected in the Tc-UBI scans of 17 patients, indicating 100% accuracy, compared with an accuracy of 90% for osteomyelitis detected in three-phase bone scans. The maximum mean A/N was observed at 15 min after intravenous injection (median: 1.91; interquartile range: 1.54-2.94). MRI was performed in 12 cases only with 75% accuracy. In addition, the A/N ratios for the Tc-UBI scans were not significantly different between patients with or without Staphylococcus aureus growth on wound cultures. CONCLUSION: For fast imaging with high accuracy, Tc-UBI 29-41 is a suitable choice for the detection of osteomyelitis. PMID- 21642883 TI - Inside intentional self-injury. PMID- 21642884 TI - Recognizing and managing rheumatoid arthritis. PMID- 21642885 TI - When you don't get the job. PMID- 21642886 TI - Menopause and the hormone controversy: clarification or confusion? AB - providers. To assist in providing women quality healthcare, recently published new guidelines help provide direction for NPs. PMID- 21642887 TI - Lenticular neovascularization subsequent to traumatic cataract formation. AB - PURPOSE: To report a series of cases involving neovascularization within the human crystalline lens-a normally avascular structure-after ocular trauma. This is a retrospective, consecutive observational case series with review of the prevailing literature. METHODS: Four individuals with a history of ocular trauma and subsequent cataract development were examined between May 2004 and April 2007. All had hypermature cataracts and intraocular inflammation, presumably secondary to phacolysis; two of the four had concurrent hyphema and ocular hypertension in the involved eye. RESULTS: All subjects in this series were found to display a discrete network of blood vessels within the structure of the crystalline lens, just beneath the anterior lens capsule. CONCLUSIONS: Neovascularization of the crystalline lens has received little attention in the ophthalmic literature, having been described only rarely in individual case reports. This manuscript details the first known case series involving lenticular neovascularization, and offers insight into its possible developmental mechanism. PMID- 21642888 TI - Knowledge and use of tear film evaluation tests by spanish practitioners. AB - PURPOSE: The present study aimed at investigating the use and knowledge of tear film and dry eye evaluation tests by a representative sample of Spanish optometrists and ophthalmologists. METHODS: A simple on-line survey was designed to explore the preferred tests for tear film evaluation and dry eye diagnosis. Additional questions surveyed knowledge of basic tear film evaluation concepts, attitude (proactive or reactive) toward patient symptoms and use of standardized dry eye questionnaires. Respondents also provided information regarding academic background, continuing education and training courses, and weekly number of tear film examinations. RESULTS: A total of 140 optometrists and 103 ophthalmologists responded to the survey. The tear break-up time test was the first preference of optometrists and ophthalmologists, whereas the Schirmer test and the non-invasive break-up time were frequently reported by ophthalmologists and optometrists, respectively, to supplement a first test. Optometrists and ophthalmologists were similar in terms of continuing education, knowledge of basic tear film concepts, and attitude regarding symptoms. Continuing education was found to positively influence knowledge, attitude toward symptoms and use of more sophisticated tear film evaluation tests. Standardized dry eye questionnaires were rarely used. A strong positive statistically significant correlation was encountered between the number of continuing education and training courses and the number of weekly tear film examinations. CONCLUSIONS: Continuing education is an invaluable tool for practitioners to increase their self-confidence and improve their clinical practice when conducting tear film evaluations and dry eye diagnosis. PMID- 21642889 TI - Variation of stereothreshold with random-dot stereogram density. AB - PURPOSE: Reports of dissimilar stereothresholds for contour and random-dot (RD) targets may reflect differences in stimulus properties or differences between local and global stereoscopic processing mechanisms. In this study, we evaluated whether the stereothresholds obtained using low- and high-density RD stimuli are consistent with a distinction between local and global disparity processing. METHODS: Stereothresholds were measured in eight normal subjects for a small disparate line segment superimposed on RD surrounds with densities that ranged between 0.07 and 28.3%. RESULTS: Stereothresholds averaged 0.23 arc min for an RD density of 0.39% and approximately doubled for lower and higher densities. The increase in stereothresholds at low densities is likely because of the increased spacing between elements, which reduces their usefulness as a reference for relative disparity judgments. The increase in stereothresholds at high densities is attributed to a crowding effect. CONCLUSIONS: Because the stereothresholds measured with RD stimuli of low and high density are limited by different constraints, they can be considered to be different types of stereotargets. However, because the stereothresholds measured for RD targets of varying densities are similar to those determined previously for a local, two-rod stereotarget, it is likely that all these stimuli are processed by a single disparity-processing mechanism. PMID- 21642890 TI - Binocular balance in normal vision and its modulation by mean luminance. AB - PURPOSE: To better understand the neural basis of sensory dominance in the normal population, we applied a recently established test designed to quantify the degree of suppression in amblyopia to participants with normal binocular vision. METHODS: This test quantifies the degree of dichoptic imbalance in coherent motion sensitivity by manipulating the contrast of stimuli seen by the two eyes. The contrast at which balanced dichoptic motion sensitivity occurs is referred to as the "balance point" and is an estimate of the degree of suppression. We apply the same logic to the measurement of sensory dominance by measuring the distribution of "balance points" within the normal population. RESULTS: We show that although most subjects are balanced or only weakly imbalanced, a minority is strongly imbalanced. To ascertain the site of sensory dominance, we assessed the degree to which normal sensory balance can be modulated by changing the interocular mean luminance. We found that mismatches in mean luminance between the two eyes had a pronounced effect on the balance point determination. CONCLUSIONS: Because cells in the lateral geniculate nucleus exhibit a strong modulation to sustained changes in the mean light level, this may suggests that the inhibitory circuits underlying sensory eye dominance are located at a precortical site. PMID- 21642892 TI - Association between polymorphisms of CYP2J2 and EPHX2 genes and risk of coronary artery disease. AB - OBJECTIVE: Common polymorphisms within cytochrome P450 2J2 (CYP2J2) and epoxide hydrolase 2 (EPHX2), which are involved in the generation or hydrolysis of epoxyeicosatrienoic acids, may determine susceptibility to the development of cardiovascular disease. To derive a more precise estimation of their relationship, we undertook a case-control study as well as a meta-analysis to assess possible associations of coronary artery disease (CAD) risk with CYP2J2 and EPHX2 genetic variations. METHODS: Associations among four single nucleotide polymorphisms in CYP2J2 and five in EPHX2 with CAD were examined in a total of 1344 cases and 1267 ethnically and geographically matched controls. To further confirm the effect of two functional variants (G-50T and R287Q) in the development of CAD, we conducted a meta-analysis including seven studies on G-50T polymorphism and six studies on R287Q polymorphism before June 2010. RESULTS: No significant association between common polymorphisms within these two genes and CAD was observed in our sample, either using methods of single-locus analysis or haplotype-based analysis. In addition, no association was detected in our meta analysis between these two functional variants and the risk of developing CAD. CONCLUSION: This case-control study as well as meta-analysis suggested no association between CYP2J2 G-50T and EPHX2 R287Q and the risk of developing CAD. PMID- 21642891 TI - Association of education and occupation with myopia in COMET parents. AB - PURPOSE: To investigate refractive error, especially myopia, in parents of myopic children and its association with education and occupation. METHODS: Six hundred twenty-seven parents (n = 375 mothers and 252 fathers) of the 469 myopic 6- to <12-year-old children enrolled in COMET provided refraction data as well as answered questions about their education and occupation. Eighty-five percent of the refractions were obtained by non-cycloplegic autorefraction (Nidek ARK 700A), and 15% were obtained from the most recent prescription. RESULTS: The mean age +/ SD of the parents was 44.26 +/- 5.81 years, and their mean spherical equivalent refraction was -2.34 +/- 2.94 D. Parents with higher education (college degree or greater) had significantly more myopia (-2.97 +/- 2.98 D) than parents with lower education (-1.72 +/- 2.76 D). The odds of being myopic were significantly higher in the higher education group (multivariate odds ratio = 2.12, 95% confidence interval = 1.41 to 3.19). Mean myopia also differed significantly by occupation, with parents in white collar jobs (-2.87 +/- 3.10 D) significantly more myopic than those in blue collar jobs (-1.21 +/- 2.02 D) by 1.66 D (p < 0.001). The odds of being myopic between the two occupation groups were of borderline significance (multivariate odds ratio = 1.61, 95% confidence interval = 0.999 to 2.60). CONCLUSIONS: The parents of myopic children participating in a clinical trial of lenses to slow the progression of myopia had a high prevalence of myopia that was associated with their level of education and to a lesser extent with their choice of occupation. To our knowledge, this is the first account of refractive errors, education, and occupation in parents of a large group of myopic children. PMID- 21642894 TI - Genetic markers anticipate response to citalopram in a majority of patients. AB - OBJECTIVE: Scientists have concluded that genetic profiles cannot predict a large percentage of variation in response to citalopram, a common antidepressant. Using the same data, we examined if a different conclusion can be arrived at when the results are personalized to fit specific patients. METHODS: We used data available through the Sequenced Treatment Alternatives to Relieve Depression database. We created three boosted Classification and Regression Trees to identify 16 subgroups of patients, among whom anticipation of positive or negative response to citalopram was significantly different from 0.5 (P<=0.1). RESULTS: In a 10-fold cross-validation, this ensemble of trees made no predictions in 33% of cases. In the remaining 67% of cases, it accurately classified response to citalopram in 78% of cases. CONCLUSION: For the majority of the patients, genetic markers can be used to guide selection of citalopram. The rules identified in this study can help personalize prescription of antidepressants. PMID- 21642893 TI - Chemotherapeutic-induced apoptosis: a phenotype for pharmacogenomics studies. AB - AIM: To determine whether cellular apoptosis is a suitable phenotypic trait for pharmacogenomics studies by evaluating caspase 3/7-mediated activity in lymphoblastoid cell lines after treatment with six chemotherapeutic agents: 5' deoxyfluorouridine, pemetrexed, cytarabine, paclitaxel, carboplatin, and cisplatin. MATERIALS AND METHODS: Using monozygotic twin pair and sibling pair lymphoblastoid cell lines, we identified conditions for measurement of caspase 3/7 activity in lymphoblastoid cell lines. Genome-wide association studies were performed with over 2 million single nucleotide polymorphisms (SNPs) and cisplatin-induced apoptosis in HapMap CEU cell lines (n=77). RESULTS: Although treatment with 5'-deoxyfluorouridine and pemetrexed for up to 24 h resulted in low levels of apoptosis or interindividual variation in caspase-dependent cell death; paclitaxel, cisplatin, carboplatin, and cytarabine treatment for 24 h resulted in 9.4-fold, 9.1-fold, 7.0-fold, and 6.0-fold increases in apoptosis relative to control, respectively. There was a weak correlation between caspase activity and cytotoxicity (r(2)=0.03-0.29) demonstrating that cytotoxicity and apoptosis are two distinct phenotypes that may produce independent genetic associations. Estimated heritability (h(2)) for apoptosis was 0.57 and 0.29 for cytarabine (5 and 40 MUmol/l, respectively), 0.22 for paclitaxel (12.5 nmol/l), and 0.34 for cisplatin (5 MUmol/l). In the genome-wide association study using the HapMap CEU panel, we identified a significant enrichment of cisplatin-induced cytotoxicity SNPs within the significant cisplatin-induced apoptosis SNPs and an enrichment of expression quantitative trait loci (eQTL). Among these eQTLs, we identified several eQTLs with known function related to apoptosis and/or cytotoxicity. CONCLUSION: Our study identifies apoptosis as a phenotype for pharmacogenomic studies in lymphoblastoid cell lines after treatment with paclitaxel, cisplatin, carboplatin, and cytarabine that may have utility for discovering biomarkers to predict response to certain chemotherapeutics. PMID- 21642895 TI - An exploration of the oligodendrocyte lineage transcription factor 2 gene and obsessive-compulsive disorder. PMID- 21642896 TI - Allelic expression imbalance of the schizophrenia susceptibility gene CHI3L1: evidence of cis-acting variation and tissue specific regulation. AB - OBJECTIVE: To identify cis-acting regulatory variants influencing the expression of the schizophrenia susceptibility gene chitinase 3-like 1 gene (CHI3L1) in human lymphoblasts and post-mortem brain tissue. METHODS: To investigate the role of cis-acting regulatory variants in controlling gene expression of CHI3L1 we quantified relative allelic abundance in individuals heterozygous for the transcribed polymorphism rs880633. Allelic quantification was performed using RNA derived from 45 individuals from the HapMap CEU panel and 41 postmortem brain samples. Association of allelic imbalance with genetic variants was determined at a gene-wide level for the HapMap samples using available genotyping data. RESULTS: Expression of the CHI3L1 transcript is under the control of potently acting cis-variation in lymphoblasts. Polymorphisms in the promoter region of CHI3L1 were significantly associated with this allelic imbalance. In the single postmortem brain tissue investigated, only moderate allelic imbalance was detected and was restricted to a small number of individuals. CONCLUSION: CHI3L1 contains common cis-acting regulatory variants that affect gene expression in lymphoblasts. A previously identified schizophrenia susceptibility variant was significantly associated with allelic imbalance in lymphoblasts. These findings do not support the notion that the schizophrenia-associated CHI3L1 variants influence gene expression in BA46 of the adult brain. We confirm that CHI3L1 contains cis-acting variation but is subject to tissue-specific regulation. PMID- 21642897 TI - Intraocular erosion and intrusion of an Arruga encircling suture. PMID- 21642898 TI - The impact of pulse duration and burn grade on size of retinal photocoagulation lesion: implications for pattern density. AB - PURPOSE: Shorter pulses used in pattern scanning photocoagulation (10-20 milliseconds [ms]) tend to produce lighter and smaller lesions than the Early Treatment Diabetic Retinopathy Study standard 100-ms exposures. Smaller lesions result in fewer complications but may potentially reduce clinical efficacy. It is worthwhile to reevaluate existing standards for the number and size of lesions needed. METHODS: The width of the coagulated zone in patients undergoing retinal photocoagulation was measured using optical coherence tomography. Lesions of "moderate," "light," and "barely visible" clinical grades were compared for 100, 200, and 400 MUm spot sizes and pulse durations of 20 ms and 100 ms. RESULTS: To maintain the same total area as in 1,000 standard burns (100 ms, moderate) with a 400-MUm beam, a larger number of 20-ms lesions are required: 1,464, 1,979, and 3,520 for moderate, light, and barely visible grades, respectively. Because of stronger relative effect of heat diffusion with a smaller beam, with 200 MUm this ratio increases: 1,932, 2,783, and 5,017 lesions of 20 ms with moderate, light, and barely visible grades correspond to the area of 1,000 standard burns. CONCLUSION: A simple formula is derived for calculation of the required spot spacing in the laser pattern for panretinal photocoagulation with various laser parameters to maintain the same total coagulated area. PMID- 21642899 TI - Early histologic and functional study of radial optic neurotomy outcomes in normal rat eyes. AB - BACKGROUND: Radial optic neurotomy (RON) has been proposed as a treatment for central retinal vein occlusion. However, it is still under debate whether RON would be an adequate treatment or a dangerous procedure, and persuasive animal studies are lacking. The aim of this study was to analyze the early histologic and functional outcomes of RON in normal rat eyes. METHODS: Radial optic neurotomy was performed by cutting into the optic nerve edge at the nasal hemisphere, while the contralateral eye underwent a sham procedure. The retinal function was assessed by scotopic electroretinography, and the visual pathway was assessed by flash visual evoked potentials. Intraocular pressure was assessed with a tonometer, the pupillary light reflex was measured after exposing eyes to a 30-second light flash, whereas the optic nerve head structure was examined by histologic analysis. RESULTS: In normal rat eyes, RON provoked minor histologic alterations at the optic nerve head level and a transient decrease in the electroretinography. No changes in visual evoked potentials, intraocular pressure, and pupillary light reflex were observed in rat eyes submitted to RON. CONCLUSION: To our knowledge, this is the first study describing the early histopathologic and functional consequences of RON in normal rat eyes. PMID- 21642900 TI - Morphologic differences in epiretinal membranes on ocular coherence tomography as a predictive factor for surgical outcome. AB - PURPOSE: To evaluate whether morphologic differences in idiopathic epiretinal membranes seen on optical coherence tomography may help predict surgical outcomes. METHODS: Seventy-five eyes of 74 patients who underwent primary pars plana vitrectomy with membrane peeling were retrospectively reviewed. Outcome measures included visual acuity, macular contour on optical coherence tomography, central macular thickness, and reoperation rate. RESULTS: According to the preoperative macular contour, 75 eyes were categorized into 4 types: 42 eyes were included in the diffuse (DIF) type, 12 in the cystoid macular edema (CME) type, 14 in the pseudolamellar hole (PLH) type, and 7 in the vitreomacular traction (VMT) type. Surgical procedure significantly improved vision in all types except for the PLH type (DIF, P < 0.0001; CME, P = 0.0378; PLH, P = 0.838; and VMT, P = 0.0273). There was a significant relationship between pre- and postoperative macular contour. All preoperative VMT showed normal contour on postoperative optical coherence tomography but had the highest reoperation rate. CONCLUSION: Surgical intervention for the PLH-type epiretinal membrane was not associated with the visual improvement seen in other epiretinal membrane types, and the VMT type had the highest reoperation rate. Future studies should evaluate the potential benefit of internal limiting membrane peeling with or without short term gas tamponade in these cases. PMID- 21642901 TI - Long-term outcomes of acute traumatic maculopathy. AB - PURPOSE: To report immediate and long-term outcomes of acute traumatic maculopathy. METHODS: Retrospective case series. Acute traumatic maculopathy was defined as a macular opacification after blunt trauma. Patients were examined at presentation, 1 week, and 6 months. Retinography and time-domain and spectral domain optical coherence tomographies were performed in all patients. Central macular thickness, the qualitative aspect of the macular profile, and retinal nerve fiber layer thickness were assessed. Multifocal electroretinography was performed at presentation and after 6 months. RESULTS: Twenty patients (20 eyes) were studied. Their mean age was 20.8 years, and the initial visual acuity was 20/100. In all cases, initial optical coherence tomographies revealed an increase in reflectivity of the inner and outer segment junction, with an apposition of the latter to the retinal pigment epithelium. Optical coherence tomography profiles were back to normal at the 1-week visit. Initial multifocal electroretinography performed in six patients showed a decrease in amplitudes in the central area but not in the periphery. There was no delay in latency. Similar electroretinal dysfunction persisted after 6 months. CONCLUSION: Macular opacification observed in acute traumatic maculopathy is associated with an increase in reflectivity of the inner and outer segment photoreceptor junction on optical coherence tomography. Although visual recovery is excellent, reduction in the electroretinal activity observed 6 months after the trauma suggests that the retina does not fully recover from the initial disorganization of its external layers. PMID- 21642902 TI - Urologic surgical simulation: an endoscopic bladder model. AB - INTRODUCTION: With the explosion of endoscopic techniques in urology as well as the increasing work restrictions with resident duty hours, training programs are faced with the challenges of how to adequately train residents while still being proficient and safe in the operating room. Surgical simulation with models is an excellent tool to help bridge the gap between practice and experience and allow residents to learn basic skills in a low stress environment that can be later transferred to the operating room. METHODS: We present a high-fidelity endoscopic boar bladder model for first-year urology resident training in preparation for real-time experience in the operating room. RESULTS: The boar bladder model held up for the residents to complete six separate tasks. In each of the six assigned tasks, both residents had a percent improvement ranging from 13% to 97% when comparing an average of the first attempts with the final attempt. CONCLUSIONS: The novel simulation model we describe demonstrates is a high-fidelity tissue surrogate that can be used for simulation training for improvement in core urologic skills by novice residents. This model may be a useful tool in documenting proficiency-based competence of cystoscopic skills. PMID- 21642903 TI - Instruction using a high-fidelity cardiopulmonary simulator improves examination skills and resource allocation in family medicine trainees. AB - INTRODUCTION: High-fidelity cardiopulmonary simulators have proven promising in various areas of medical education but have yet to be studied in Family Medicine training. METHODS: A 2-hour curriculum, combining didactic and simulator exposure, and addressing common valvular pathologies, was offered to post graduate year 1 and 2 Family Medicine residents. Residents' abilities to describe and diagnose four simulated murmurs were assessed before the teaching sessions and 2 to 4 weeks after. Confidence in physical examination skills, as well as the use of echocardiography, was also measured. RESULTS: Twenty residents participated. Mean composite murmur description scores improved in 95% of residents (P < 0.001), as did mean diagnostic accuracy (from 43.8% to 85.0%; P < 0.001). For pathologic murmurs, the number of echocardiograms recommended did not change, whereas for the nonpathologic murmur, 16 residents who recommended echocardiography presession no longer did postsession (P < 0.001). Mean confidence significantly increased (P < 0.001). The mean satisfaction score for the session was 4.9/5, and all residents recommended that the session be repeated in future years. CONCLUSION: A didactic and simulator-based session is very well received by Family Medicine residents. It significantly improves description and diagnosis of murmurs and reduces unnecessary echocardiogram use without affecting appropriate use. PMID- 21642904 TI - Nontechnical skills assessment after simulation-based continuing medical education. AB - INTRODUCTION: Human factors have been identified as root causes of human error in medicine. The "Anesthetists' Non-Technical Skills (ANTS) system" evaluates the effect of simulation training and debriefing on nontechnical skills (NTS). Studies suggest that residents' NTS may improve after simulation training but the effect on NTS of practicing anesthesiologists is unclear. The purpose of this study was to determine whether high-fidelity simulation training and debriefing improved the NTS of practicing anesthesiologists using the ANTS tool. METHODS: In a previous study, 67 practicing anesthesiologists managed a 45-minute standardized anesthetic case using high-fidelity simulation and returned 5 to 9 months later to manage a second case. After Research Ethics Board approval, two blinded video reviewers, trained in the use of the ANTS system, evaluated archived videotapes of the 59 subjects who completed both sessions. Results were analyzed with a mixed-design analysis of variance. Interrater reliability was calculated using the intraclass correlation coefficient. RESULTS: Interrater reliability for the ANTS scoring was 0.436, P < 0.05. Overall, ANTS scores improved approximately 5% from session 1 to 2 (P < 0.01), but there was no effect due to debriefing. The situational awareness ANTS category showed a statistically significant effect of debriefing (P < 0.05). CONCLUSIONS: The relatively short simulation intervention, the length of time until the posttest was completed, well-developed NTS in practicing physicians, and a tool that might not be the optimal method of measurement may all account for the lack of improvement in NTS of practicing anesthesiologists as demonstrated in this study. PMID- 21642905 TI - Impact of simulation-based extracorporeal membrane oxygenation training in the simulation laboratory and clinical environment. AB - INTRODUCTION: Extracorporeal membrane oxygenation (ECMO) is a high-risk, complex therapy. Opportunities to develop teamwork skills and expertise to mitigate risks are few. Our objective was to assess whether simulation would improve technical and nontechnical skills in dealing with ECMO circuit emergencies and allow transfer of skills from the simulated setting to clinical environment. METHODS: Subjects were ECMO circuit providers who performed scenarios utilizing an infant simulator and functional ECMO circuit, followed immediately by video-assisted debriefings. Within the simulation laboratory, outcomes were timed responses, percentage of correct actions, teamwork, safety knowledge, and attitudes. Identification of latent safety threats (LSTs) was the focus of debriefings. Within the clinical setting, translation of learned skills was assessed by measuring circuit readiness and compliance with a cannulation initiation checklist. RESULTS: Nineteen subjects performed 96 simulations during enrollment. In the laboratory, there was no improvement in timed responses or percent correct actions. Teamwork (P = 0.001), knowledge (P = 0.033), and attitudes (P = 0.001) all improved compared with baseline. Debriefing identified 99 LSTs. Clinically, 26 cannulations occurred during enrollment. Median time from blood available to circuit readiness was 17 minutes (range, 5-95), with no improvement during the study. Compliance with the initiation checklist improved compared with prestudy baseline (P < 0.0001). CONCLUSIONS: Simulation-based training is an effective method to improve safety knowledge, attitudes, and teamwork surrounding ECMO emergencies. On-going training is feasible and allows identification of LSTs. Further work is needed to assess translation of learned skills and behaviors into the clinical environment. PMID- 21642906 TI - Design and evaluation of simulation scenarios for a program introducing patient safety, teamwork, safety leadership, and simulation to healthcare leaders and managers. AB - We developed a training program to introduce managers and informal leaders of healthcare organizations to key concepts of teamwork, safety leadership, and simulation to motivate them to act as leaders to improve safety within their sphere of influence. This report describes the simulation scenario and debriefing that are core elements of that program. Twelve teams of clinician and nonclinician managers were selected from a larger set of volunteers to participate in a 1-day, multielement training program. Two simulation exercises were developed: one for teams of nonclinicians and the other for clinicians or mixed groups. The scenarios represented two different clinical situations, each designed to engage participants in discussions of their safety leadership and teamwork issues immediately after the experience. In the scenarios for nonclinicians, participants conducted an anesthetic induction and then managed an ethical situation. The scenario for clinicians simulated a consulting visit to an emergency room that evolved into a problem-solving challenge. Participants in this scenario had a limited time to prepare advice for hospital leadership on how to improve observed safety and cultural deficiencies. Debriefings after both types of scenarios were conducted using principles of "debriefing with good judgment." We assessed the relevance and impact of the program by analyzing participant reactions to the simulation through transcript data and facilitator observations as well as a postcourse questionnaire. The teams generally reported positive perceptions of the relevance and quality of the simulation with varying types and degrees of impact on their leadership and teamwork behaviors. These kinds of clinical simulation exercises can be used to teach healthcare leaders and managers safety leadership and teamwork skills and behaviors. PMID- 21642907 TI - Long-term outcome in kidney transplant recipients over 70 years in the Eurotransplant Senior Kidney Transplant Program: a single center experience. AB - BACKGROUND: Kidney transplantation in the elderly is complicated by comorbidities and a higher incidence of death. The Eurotransplant Senior Program (ESP) has been established to allocate kidneys from older donors to the increasing number of older recipients. In this retrospective, single center data analysis, we compare the outcome of recipients older than 70 years with younger recipients transplanted under the ESP protocol. METHODS: Between 1999 and 2009, a total of 83 kidneys were transplanted under the ESP protocol in Innsbruck and 19 of the recipients were older than 70 years (mean, 72.7 years). Cold ischemia time was kept short in both groups by giving preference to regional donor organs. RESULTS: Patient survival at 1 and 5 years were 95% and 67% in the 70+ group and 94.4% and 82.6% in the 70- group. Graft survival was 95% and 52% at 1 and 5 years in the 70+ group and 94.4% and 79.0% in the 70- group. When censored for death, graft survival at year 1 and 5 were 100% and 82% in the 70+ group and 98.1% and 92.7% in the 70- group. The delayed graft function rate was high in both groups (36.8% and 41.1%, respectively). Morbidities were largely related to hemodynamic, oncologic, and infectious events. Cardiac failure was the major cause of death. CONCLUSION: Relatively good results can be achieved with renal transplantation in patients older than 70 years under careful pretransplant evaluation and postoperative management of comorbidities. PMID- 21642908 TI - The sound of thunder. PMID- 21642909 TI - 'Care and compassion': why does it go wrong? PMID- 21642910 TI - Patients with inflammatory arthritis: an opportunity for community nurses. AB - People with Inflammatory joint disease, e.g. rheumatoid arthritis (RA), have traditionally been treated and managed within secondary care on an outpatient basis. This group of patients has varied and complex needs. With increasing emphasis on offering care closer to home, there is an opportunity for nurses based in the community to become more involved in the management and support of these patients. The level of involvement will depend on the resources available locally and will require education and support for the role. Based on recommendations in recent publications and the author's personal experience working in three different rheumatology departments, this article explores the ways in which community nurses can support people with RA in the community and identify sources for further professional development to support and maintain this. PMID- 21642911 TI - Nurses' needs in delivering palliative care for long-term conditions. AB - This study addressed the question 'What are the needs of community nurses in delivering palliative care to people with long-term conditions?' A qualitative exploratory descriptive design was employed. Ten community nurses (Band 5-7) were recruited from a purposive sample following a process of randomised stratified sampling, according to geographical area and Band for matched numbers. Semi structured interviews were undertaken and audio recorded with written informed consent. The interviews were transcribed verbatim and analysed using an adapted Burnard's framework. The study found that establishing therapeutic relationships, having access to resources, co-ordination and provision of clinical care and collaborative working were all highlighted by the community nurses as needs in delivering palliative care. If these four needs were met, the community nurses believed they could deliver palliative care to their patients. Issues around a lack of resources, community nurses' educational needs and the late referral of patients with non-malignant long-term conditions to community nursing were also identified. PMID- 21642912 TI - Infection control in the community: a pragmatic approach. AB - Infection can have a detrimental and potentially life-threatening impact on the health and wellbeing of patients. Infection control and prevention is as important in the community as it is in an acute hospital. This article summarizes the key infection and prevention issues in community nursing. It offers a pragmatic approach as community settings may be challenging to both infection control and community healthcare professionals. Patient safety is the top priority, and ensuring that safe practices are followed to reduce the risk of infection regardless of healthcare setting is paramount. There are many external factors that may impede infection control practices when delivering care in a patient's own home, and this article sets out the factors that must be considered in order to manage the risk. PMID- 21642913 TI - Expert patients: the Service User Mentorship Programme. AB - As part of its remit to improve the lives of people with long-term conditions, the Expert Patients Programme Community Interest Company has partnered with South Central Strategic Health Authority and a number of local universities to provide Service User Mentors for students on the Foundation Degree in Health and Social Care Long Term Conditions Pathway. This article discusses how giving students the opportunity to have greater insight into what it means to live with a chronic condition, and an understanding of and improved practice in effective self management, can support and improve health and social care outcomes for people living with long-term conditions. PMID- 21642914 TI - Community nursing in systems reform: the London polyclinic experience. AB - Internationally, the polyclinic has been a feature of many health systems. The recent UK policy shift towards enhanced coordination of care closer to home resulted in the development of polyclinics, most notably in London. This article explores the background to the development of polyclinics and draws on the early experience of developments in London to explore what their impact has been, and is likely to be, on community nursing. Emerging findings from an evaluation of four pilot polyclinics suggests that rather than one model, polyclinics evolved in distinctively different ways more appropriately labelled as polysystems. Although policy makers clearly identified community nursing as being one of the key components of integrated, community-based care, the evaluation suggests a focus on high-level organizational restructuring and system change can shift attention from what many would consider core activities such as community health services. PMID- 21642915 TI - Falls prevention among the elderly: an update from Age UK. PMID- 21642916 TI - Lindsay Leg Club Foundation agrees charity partnership with RAFT. PMID- 21642917 TI - Doing more with less. PMID- 21642924 TI - The ripple effect in community nursing. PMID- 21642925 TI - Use of emollients in the treatment of dry skin conditions. AB - Dry skin conditions are often not considered important by health professionals, and as a consequence treatments are frequently under-prescribed. As a result of this, problems with untreated dry skin can lead to a variety of issues. Conditions such as pruritus and ichthyosis vulgaris can be distressing, while ezcema and psoriasis can lead to more serious consequences such as fissures and infections and can result in a reduced quality of life and social isolation. However, with appropriate treatments such as emollient therapy, the incidences of these conditions occurring can be reduced. Despite a wide range of choice available on the market, choosing an effective emollient which will help aid concordance is often difficult. In order to achieve this, several factors need to be considered in conjunction with the product selection. This article discusses the various factors that nurses need to consider when advising patients on which emollient to use as part of their skin care regimen, in order to improve concordance and patient outcomes. PMID- 21642926 TI - Risk assessment and management tools for constipation. AB - Constipation is a distressing condition that can happen to anyone, but is of particular concern in the elderly and less mobile, people with neurological impairments, and those in institutional care. The causes of constipation are multifactorial, and can be influenced by physical, psychological, physiological, emotional and environmental factors. This article discusses definitions, symptoms and types of constipation, and provides guidance on risk factors and assessment using the Norgine Risk Assessment Tool for Constipation. PMID- 21642927 TI - Developing a structured education programme for clients with COPD. AB - This article describes the development of a Structured Education Pulmonary Rehabilitation Programme (SEPRP), for the PRINCE (Pulmonary Rehabilitation In Nurse-led Community Environments) study. This process involved a literature review of existing PR programmes, a content analysis of national and international PR programmes and a concept analysis of empowerment. Secondly, two small descriptive qualitative studies were undertaken to further inform programme content exploring the views of health professionals and clients on programme content, as well as their perceptions of living with COPD. The findings of these two stages led to the development of the eight-week PRINCE SEPRP, based on a philosophy of empowerment and the SEPRP was underpinned by the Transtheoretical model (TTM) of behaviour change. Programme content included managing medications and managing breathlessness, knowing and managing symptoms, recognizing and managing acute exacerbations as well as an individualized exercise programme. Participants' experiences of self management following participation in the SEPRP were also examined. The findings indicate that nurses working in primary care have an important role in informing the development of SEPRP as well as having a key role in their delivery. They have the potential to work in empowering ways with COPD clients and the PRINCE SEPRP is just one example of how this can be put into practice. PMID- 21642928 TI - Identifying care actions to conserve dignity in end-of-life care. AB - Community nurses have a central role in the provision of palliative and end-of life care; helping people to die with dignity is an important component of this care. To conserve dignity, care should comprise a broad range of actions addressing the distress that might impact on the patient's sense of dignity. These care actions need to be defined. This study aims to suggest care actions that conserve dignity at the end of life based on evidence from local experience and community nursing practice. Data were collected by focus group interviews and analysed by framework analysis using the Chochinov model of dignity as a predefined framework. Suggestions on care actions were given in relation to all themes. As part of a multi-phase project developing and testing a dignity care pathway, this study might help community nurses to conserve dying patients' dignity. PMID- 21642929 TI - Taking hypoglycaemia seriously: diabetes, dementia and heart disease. AB - Hypoglycaemia is a common and potentially deadly problem for people with diabetes who are prescribed insulin secretagogues and/or insulin as part of their treatment regimen. This article focuses mainly on type 2 diabetes and considers the incidence, pharmacological causes and definition of hypoglycaemia. It discusses the recently defined relationships between hypoglycaemia and the onset and development of dementia and cardiovascular disease. Hypoglycaemia risk factors are identified and linked to specific characteristics found in the elderly, and information is offered on how to treat hypoglycaemia effectively and avoid repeated episodes. PMID- 21642930 TI - Determining a patient's best interests. AB - Best interests is the standard by which the law judges the propriety of care and treatment for adults who lack decision-making capacity. Since the introduction of mental capacity legislation, determining best interests has moved from a paternalistic risk benefit analysis by the district nurse providing care to an incapable patient to a more holistic approach where the wishes, views and values of the patient and their relatives must be considered. This article discusses how district nurses should approach the assessment and determination of best interests to meet the requirements of the law. PMID- 21642931 TI - Communicating and coping: essential end-of-life care skills. AB - The QNI has introduced a palliative care award for nurses. Rosemary Cook explains the origin of the award and why Angela Parle was chosen to receive it. PMID- 21642932 TI - Nurses with special interests. AB - High Quality Care for All (Department of Health (DH), 2008) emphasised the centrality of quality within NHS provision based upon a service that is 'fair, personalized, effective and safe'. The challenge of meeting this aspiration is even greater today amid resource constraints and the imperative of transferring more complex care out of hospitals and into primary care and community settings. The quality of care delivered to older people in particular has attracted much negative comment (Ombudsman, 2011), with the values of care and compassion appearing to have evaporated in some services. PMID- 21642933 TI - Altholactone displays promising antimicrobial activity. AB - The antimicrobial activity of altholactone, a naturally extracted styryllactone isolated from Goniothalamus malayanus, was determined against Gram positive (S. aureus ATTC 25923, S. aureus ATTC 25392, and E. faecalis ATTC 29212) and Gram negative (E. coli ATTC 35218, S. typhi ATTC 14023 and P. aeruginosa ATCC 27853) reference bacteria and against the fungus C. albicans ATTC 10231. Different concentrations of altholactone (0, 12, 25, and 50 MUg/mL) were used. Results revealed that altholactone inhibited the growth of all tested microbes except P. aeruginosa ATCC 27853 in a dose-dependent manner, with the highest cytotoxic effects occurring at 50 MUg/mL. The average of the inhibition zones of the different concentrations was between 0-30 mm. Furthermore, altholactone-induced antimicrobial activity against the more sensitive microbes was assessed by measuring the minimal inhibitory concentration (MIC). Results indicated that Gram positive (S. aureus ATTC 25923, S. aureus ATTC 25392, and E. faecalis ATTC 29212) cells were more sensitive to altholactone than Gram negative ones (E. coli ATTC 35218, S. typhi ATTC 14023). C. albicans showed moderate sensitivity. These results indicate that altholactone might be a potential antimicrobial agent, particularly in ciprofloxacin-refractory S. aureus and E. faecalis infections. Further investigations are required to illustrate the mechanism(s) by which altholactone produces its antimicrobial effects. PMID- 21642934 TI - Curcumin: an anti-inflammatory molecule from a curry spice on the path to cancer treatment. AB - Oxidative damage and inflammation have been pointed out in preclinical studies as the root cause of cancer and other chronic diseases such as diabetes, hypertension, Alzheimer's disease, etc. Epidemiological and clinical studies have suggested that cancer could be prevented or significantly reduced by treatment with anti-oxidant and anti-inflammatory drugs, therefore, curcumin, a principal component of turmeric (a curry spice) showing strong anti-oxidant and anti inflammatory activities, might be a potential candidate for the prevention and/or treatment of cancer and other chronic diseases. However, curcumin, a highly pleiotropic molecule with an excellent safety profile targeting multiple diseases with strong evidence on the molecular level, could not achieve its optimum therapeutic outcome in past clinical trials, largely due to its low solubility and poor bioavailability. Curcumin can be developed as a therapeutic drug through improvement in formulation properties or delivery systems, enabling its enhanced absorption and cellular uptake. This review mainly focuses on the anti inflammatory potential of curcumin and recent developments in dosage form and nanoparticulate delivery systems with the possibilities of therapeutic application of curcumin for the prevention and/or treatment of cancer. PMID- 21642935 TI - Fluorescent probes detecting the phagocytic phase of apoptosis: enzyme-substrate complexes of topoisomerase and DNA. AB - In apoptosis, the initial self-driven suicide phase generates cellular corpses which are digested in the phagolysosomes of professional and amateur phagocytes during the subsequent waste-management phase. This ensures the complete elimination of the genetic material which often contains pathological, viral or cancerous DNA sequences. Although the phagocytic phase is critical for the efficient execution of apoptosis, there are currently few methods specifically adapted for its detailed visualization in the fixed tissue section format. To resolve this we developed new fluorescent probes for in situ research. The probes selectively visualize active phagocytic cells of any lineage (professional, amateur phagocytes or surrounding tissue cells) which engulf and digest apoptotic cell DNA. These fluorescent probes are the covalently-bound enzyme-DNA intermediates produced in a topoisomerase reaction with specific "starting" oligonucleotides. They detect a specific marker of DNase II cleavage activity, which occurs exclusively in phagolysosomes of the cells that engulfed apoptotic nuclei. The probes provide snap-shot images of the digestion process occurring in cellular organelles responsible for the actual execution of phagocytic degradation of apoptotic cell corpses. We applied the probes for visualization of the phagocytic reaction in tissue sections of normal thymus and in several human lymphomas. We also discuss the nature, stability and properties of DNase II-type breaks as a marker of phagocytic activity. This development provides a useful fluorescent tool for studies of pathologies where clearance of dying cells is essential, such as cancers, inflammation, infection and auto-immune disorders. PMID- 21642936 TI - A high molar extinction coefficient mono-anthracenyl bipyridyl heteroleptic ruthenium(II) complex: synthesis, photophysical and electrochemical properties. AB - In our quest to develop good materials as photosensitizers for photovoltaic dye sensitized solar cells (DSSCs), cis-dithiocyanato-4-(2,3-dimethylacrylic acid) 2,2'-bipyridyl-4-(9-anthracenyl-(2,3-dimethylacrylic)-2,2'-bipyridyl ruthenium(II) complex, a high molar extinction coefficient charge transfer sensitizer, was designed, synthesized and characterized by spectroscopy and electrochemical techniques. Earlier studies on heteroleptic ruthenium(II) complex analogues containing functionalized oligo-anthracenyl phenanthroline ligands have been reported and documented. Based on a general linear correlation between increase in the length of pi-conjugation bond and the molar extinction coefficients, herein, we report the photophysical and electrochemical properties of a Ru(II) bipyridyl complex analogue with a single functionalized anthracenyl unit. Interestingly, the complex shows better broad and intense metal-to ligand charge transfer (MLCT) band absorption with higher molar extinction coefficient (lambda(max) = 518 nm, e = 44900 M-1 cm-1), and appreciable photoluminescence spanning the visible region than those containing higher anthracenyl units. It was shown that molar absorption coefficient of the complexes may not be solely depended on the extended pi-conjugation but are reduced by molecular aggregation in the molecules. PMID- 21642937 TI - New catechol derivatives of safrole and their antiproliferative activity towards breast cancer cells. AB - Catechols were synthesized from safrole. Nine derivatives were prepared and assessed for antiproliferative effects using different human cell lines. The in vitro growth inhibition assay was based on the sulphorhodamine dye to quantify cell viability. The derivatives 4-allylbenzene-1,2-diol (3), 4 4-[3 (acetyloxy)propyl]-1,2-phenylene diacetate (6) and 4-[3-(acetyloxy)propyl]-5 nitro-1,2-phenylene diacetate (10) showed higher cytotoxicity than the parent compound 2 in tests performed on two breast cancer cell lines (MCF-7 and MDA-MB 231). The IC50 values of 40.2 +/- 6.9 MUM, 5.9 +/- 0.8 MUM and 33.8 +/- 4.9 MUM, respectively, were obtained without toxicity towards dermal human fibroblast (DHF cells). PMID- 21642938 TI - Triterpenoids from the roots of Sanguisorba tenuifolia var. Alba. AB - The ethyl acetate soluble fraction from the roots of Sanguisorba tenuifolia was found to have a hypoglucemic effect in alloxan-induced diabetic rats. Two new triterpenoids, identified as 2-oxo-3beta,19alpha-dihydroxyolean-12-en-28-oic acid beta-D-gluco-pyranosyl ester (1) and 2alpha,19alpha-dihydroxy-3-oxo-12-ursen-28 oic acid beta-D-glucopyranosyl ester (4) were isolated from this fraction, along with thirteen known triterpenoids. Their structures were elucidated by chemical and spectroscopic methods. All these compounds demonstrated inhibitory activities against alpha-glucosidase with IC50 values in the 0.62-3.62 mM range. PMID- 21642939 TI - On the pi coordination of organometallic fullerene complexes. AB - Novel organometallic complexes of fullerene C80 and aryl ligands were simulated. The nature and characteristics of this family of complexes involving pi coordination between the fullerene and a metal centre have been studied from a theoretical point of view. We are particularly interested in complexes where eta6 coordination is present, this being the strangest manifestation of known coordinations, and thus we have studied several known and simulated compounds of this kind in order to understand the lack of examples. The presence of other eta6 or eta5 ligands on the opposite side seems to be an important element aiding the stabilization of these complexes, also inducing the conductive and semiconductive behaviour of the studied species. PMID- 21642940 TI - Synthesis and antifungal evaluation of 1-aryl-2-dimethyl- aminomethyl-2-propen-1 one hydrochlorides. AB - The development of resistance to current antifungal therapeutics drives the search for new effective agents. The fact that several acetophenone-derived Mannich bases had shown remarkable antifungal activities in our previous studies led us to design and synthesize some acetophenone-derived Mannich bases, 1-8 and 2-acetylthiophene-derived Mannich base 9, 1-aryl-2-dimethylaminomethyl-2-propen-1 one hydrochloride, to evaluate their antifungal activities. The designed chemical structures have alpha,beta-unsaturated ketone moieties, which are responsible for the bioactivities of the Mannich bases. The aryl part was C6H5(1); 4-CH3C6H4 (2); 4-CH3OC6H4 (3); 4-ClC6H4 (4); 4-FC6H4 (5); 4-BrC6H4 (6); 4-HOC6H4 (7); 4-NO2C6H4 (8); and C4H3S(2-yl) (9). In this study the designed compounds were synthesized by the conventional heating method and also by the microwave irradiation method to compare these methods in terms of reaction times and yields to find an optimum synthetic method, which can be applied for the synthesis of Mannich bases in further studies. Since there are limited number of studies reporting the synthesis of Mannich bases by microwave irradiation, this study may also contribute to the general literature on Mannich bases. Compound 7 was reported for the first time. Antifungal activities of all compounds and synthesis of the compounds by microwave irradiation were also reported for the first time by this study. Fungi (15 species) were used for antifungal activity test. Amphotericin B was tested as an antifungal reference compound. In conclusion, compounds 1-6, and 9, which had more potent (2-16 times) antifungal activity than the reference compound amphotericin B against some fungi, can be model compounds for further studies to develop new antifungal agents. In addition, microwave irradiation can be considered to reduce reaction period, while the conventional method can still be considered to obtain compounds with higher reaction yields in the synthesis of new Mannich bases. PMID- 21642941 TI - Determining optimum conditions for lipase-catalyzed synthesis of triethanolamine (TEA)-based esterquat cationic surfactant by a Taguchi robust design method. AB - A Taguchi robust design method with an L9 orthogonal array was implemented to optimize experimental conditions for the biosynthesis of triethanolamine (TEA) based esterquat cationic surfactants using an enzymatic reaction method. The esterification reaction conversion% was considered as the response. Enzyme amount, reaction time, reaction temperature and molar ratio of substrates, [oleic acid: triethanolamine (OA:TEA)] were chosen as main parameters. As a result of the Taguchi analysis in this study, the molar ratio of substrates was found to be the most influential parameter on the esterification reaction conversion%. The amount of enzyme in the reaction had also a significant effect on reaction conversion%. PMID- 21642942 TI - Channelopathies and drug discovery in the postgenomic era. PMID- 21642943 TI - Physiological roles and diseases of Tmem16/Anoctamin proteins: are they all chloride channels? AB - The Tmem16 gene family was first identified by bioinformatic analysis in 2004. In 2008, it was shown independently by 3 laboratories that the first two members (Tmem16A and Tmem16B) of this 10-gene family are Ca(2+)-activated Cl(-) channels. Because these proteins are thought to have 8 transmembrane domains and be anion selective channels, the alternative name, Anoctamin (anion and octa=8), has been proposed. However, it remains unclear whether all members of this family are, in fact, anion channels or have the same 8-transmembrane domain topology. Since 2008, there have been nearly 100 papers published on this gene family. The excitement about Tmem16 proteins has been enhanced by the finding that Ano1 has been linked to cancer, mutations in Ano5 are linked to several forms of muscular dystrophy (LGMDL2 and MMD-3), mutations in Ano10 are linked to autosomal recessive spinocerebellar ataxia, and mutations in Ano6 are linked to Scott syndrome, a rare bleeding disorder. Here we review some of the recent developments in understanding the physiology and structure-function of the Tmem16 gene family. PMID- 21642944 TI - Targeting F508del-CFTR to develop rational new therapies for cystic fibrosis. AB - The mutation F508del is the commonest cause of the genetic disease cystic fibrosis (CF). CF disrupts the function of many organs in the body, most notably the lungs, by perturbing salt and water transport across epithelial surfaces. F508del causes harm in two principal ways. First, the mutation prevents delivery of the cystic fibrosis transmembrane conductance regulator (CFTR) to its correct cellular location, the apical (lumen-facing) membrane of epithelial cells. Second, F508del perturbs the Cl(-) channel function of CFTR by disrupting channel gating. Here, we discuss the development of rational new therapies for CF that target F508del-CFTR. We highlight how structural studies provide new insight into the role of F508 in the regulation of channel gating by cycles of ATP binding and hydrolysis. We emphasize the use of high-throughput screening to identify lead compounds for therapy development. These compounds include CFTR correctors that restore the expression of F508del-CFTR at the apical membrane of epithelial cells and CFTR potentiators that rescue the F508del-CFTR gating defect. Initial results from clinical trials of CFTR correctors and potentiators augur well for the development of small molecule therapies that target the root cause of CF: mutations in CFTR. PMID- 21642945 TI - Calcium binding protein-mediated regulation of voltage-gated calcium channels linked to human diseases. AB - Calcium ion entry through voltage-gated calcium channels is essential for cellular signalling in a wide variety of cells and multiple physiological processes. Perturbations of voltage-gated calcium channel function can lead to pathophysiological consequences. Calcium binding proteins serve as calcium sensors and regulate the calcium channel properties via feedback mechanisms. This review highlights the current evidences of calcium binding protein-mediated channel regulation in human diseases. PMID- 21642946 TI - Targeting ryanodine receptors for anti-arrhythmic therapy. AB - Antiarrhythmic drugs are a group of pharmaceuticals that suppress or prevent abnormal heart rhythms, which are often associated with substantial morbidity and mortality. Current antiarrhythmic drugs that typically target plasma membrane ion channels have limited clinical success and in some cases have been described as being pro-arrhythmic. However, recent studies suggest that pathological release of calcium (Ca(2+)) from the sarcoplasmic reticulum via cardiac ryanodine receptors (RyR2) could represent a promising target for antiarrhythmic therapy. Diastolic SR Ca(2+) release has been linked to arrhythmogenesis in both the inherited arrhythmia syndrome 'catecholaminergic polymorphic ventricular tachycardia' and acquired forms of heart disease (eg, atrial fibrillation, heart failure). Several classes of pharmaceuticals have been shown to reduce abnormal RyR2 activity and may confer protection against triggered arrhythmias through reduction of SR Ca(2+) leak. In this review, we will evaluate the current pharmacological methods for stabilizing RyR2 and suggest treatment modalities based on current evidence of molecular mechanisms. PMID- 21642947 TI - A role of stretch-activated potassium currents in the regulation of uterine smooth muscle contraction. AB - Rates of premature birth are alarming and threaten societies and healthcare systems worldwide. Premature labor results in premature birth in over 50% of cases. Preterm birth accounts for three-quarters of infant morbidity and mortality. Children that survive birth before 34 weeks gestation often face life long disability. Current treatments for preterm labor are wanting. No treatment has been found to be generally effective and none are systematically evaluated beyond 48 h. New approaches to the treatment of preterm labor are desperately needed. Recent studies from our laboratory suggest that the uterine muscle is a unique compartment with regulation of uterine relaxation unlike that of other smooth muscles. Here we discuss recent evidence that the mechanically activated 2 pore potassium channel, TREK-1, may contribute to contraction-relaxation signaling in uterine smooth muscle and that TREK-1 gene variants associated with human labor and preterm labor may lead to a better understanding of preterm labor and its possible prevention. PMID- 21642948 TI - Beyond membrane channelopathies: alternative mechanisms underlying complex human disease. AB - Over the past fifteen years, our understanding of the molecular mechanisms underlying human disease has flourished in large part due to the discovery of gene mutations linked with membrane ion channels and transporters. In fact, ion channel defects ("channelopathies" - the focus of this review series) have been associated with a spectrum of serious human disease phenotypes including cystic fibrosis, cardiac arrhythmia, diabetes, skeletal muscle defects, and neurological disorders. However, we now know that human disease, particularly excitable cell disease, may be caused by defects in non-ion channel polypeptides including in cellular components residing well beneath the plasma membrane. For example, over the past few years, a new class of potentially fatal cardiac arrhythmias has been linked with cytoplasmic proteins that include sub-membrane adapters such as ankyrin-B (ANK2), ankyrin-G (ANK3), and alpha-1 syntrophin, membrane coat proteins including caveolin-3 (CAV3), signaling platforms including yotiao (AKAP9), and cardiac enzymes (GPD1L). The focus of this review is to detail the exciting role of lamins, yet another class of gene products that have provided elegant new insight into human disease. PMID- 21642949 TI - Drug discovery for polycystic kidney disease. AB - In polycystic kidney disease (PKD), a most common human genetic diseases, fluid filled cysts displace normal renal tubules and cause end-stage renal failure. PKD is a serious and costly disorder. There is no available therapy that prevents or slows down the cystogenesis and cyst expansion in PKD. Numerous efforts have been made to find drug targets and the candidate drugs to treat PKD. Recent studies have defined the mechanisms underlying PKD and new therapies directed toward them. In this review article, we summarize the pathogenesis of PKD, possible drug targets, available PKD models for screening and evaluating new drugs as well as candidate drugs that are being developed. PMID- 21642950 TI - Propofol and arrhythmias: two sides of the coin. AB - The hypnotic agent propofol is effective for the induction and maintenance of anesthesia. However, recent studies have shown that propofol administration is related to arrhythmias. Propofol displays both pro- and anti-arrhythmic effects in a concentration-dependent manner. Data indicate that propofol can convert supraventricular tachycardia and ventricular tachycardia and may inhibit the conduction system of the heart. The mechanism of the cardiac effects remains poorly defined and may involve ion channels, the autonomic nervous system and cardiac gap junctions. Specifically, sodium, calcium and potassium currents in cardiac cells are suppressed by clinically relevant concentrations of propofol. Propofol shortens the action potential duration (APD) but lessens the ischemia induced decrease in the APD. Furthermore, propofol suppresses both sympathetic and parasympathetic tone and preserves gap junctions during ischemia. All of these effects cumulatively contribute to the antiarrhythmic and proarrhythmic properties of propofol. PMID- 21642951 TI - Characterization of a critical role for CFTR chloride channels in cardioprotection against ischemia/reperfusion injury. AB - AIM: To further characterize the functional role of cystic fibrosis transmembrane conductance regulator (CFTR) in early and late (second window) ischemic preconditioning (IPC)- and postconditioning (POC)-mediated cardioprotection against ischemia/reperfusion (I/R) injury. METHODS: CFTR knockout (CFTR(-/-)) mice and age- and gender-matched wild-type (CFTR(+/+)) and heterozygous (CFTR(+/ )) mice were used. In in vivo studies, the animals were subjected to a 30-min coronary occlusion followed by a 40-min reperfusion. In ex vivo (isolate heart) studies, a 45-min global ischemia was applied. To evaluate apoptosis, the level of activated caspase 3 and TdT-mediated dUTP-X nick end labeling (TUNEL) were examined. RESULTS: In the in vivo I/R models, early IPC significantly reduced the myocardial infarct size in wild-type (CFTR(+/+)) (from 40.4% +/- 5.3% to 10.4% +/ 2.0%, n=8, P<0.001) and heterozygous (CFTR(+/-)) littermates (from 39.4% +/- 2.4% to 15.4% +/- 5.1%, n=6, P<0.001) but failed to protect CFTR knockout (CFTR( /-)) mice from I/R induced myocardial infarction (46.9% +/- 6.2% vs 55.5% +/- 7.8%, n=6, P>0.5). Similar results were observed in the in vivo late IPC experiments. Furthermore, in both in vivo and ex vivo I/R models, POC significantly reduced myocardial infarction in wild-type mice, but not in CFTR knockout mice. In ex vivo I/R models, targeted inactivation of CFTR gene abolished the protective effects of IPC against I/R-induced apoptosis. CONCLUSION: These results provide compelling evidence for a critical role for CFTR Cl(-) channels in IPC- and POC-mediated cardioprotection against I/R-induced myocardial injury. PMID- 21642952 TI - Enhanced salt sensitivity following shRNA silencing of neuronal TRPV1 in rat spinal cord. AB - AIM: To investigate the effects of selective knockdown of TRPV1 channels in the lower thoracic and upper lumbar segments of spinal cord, dorsal root ganglia (DRG) and mesenteric arteries on rat blood pressure responses to high salt intake. METHODS: TRPV1 short-hairpin RNA (shRNA) was delivered using intrathecal injection (6 MUg . kg(-1) . d(-1), for 3 d). Levels of TRPV1 and tyrosine hydroxylase expression were determined by Western blot analysis. Systolic blood pressure and mean arterial pressure (MAP) were examined using tail-cuff and direct arterial measurement, respectively. RESULTS: In rats injected with control shRNA, high-salt diet (HS) caused higher systolic blood pressure compared with normal-salt diet (NS) (HS:149 +/- 4 mmHg; NS:126 +/- 2 mmHg, P<0.05). Intrathecal injection of TRPV1 shRNA significantly increased the systolic blood pressure in both HS rats and NS rats (HS:169 +/- 3 mmHg; NS:139 +/- 2 mmHg). The increases was greater in HS rats than in NS rats (HS: 13.9% +/- 1.8%; NS: 9.8 +/- 0.7, P<0.05). After TRPV1 shRNA treatment, TRPV1 expression in the dorsal horn and DRG of T8-L3 segments and in mesenteric arteries was knocked down to a greater extent in HS rats compared with NS rats. Blockade of alpha1-adrenoceptors abolished the TRPV1 shRNA-induced pressor effects. In rats injected with TRPV1 shRNA, level of tyrosine hydroxylase in mesenteric arteries was increased to a greater extent in HS rats compared with NS rats. CONCLUSION: Selective knockdown of TRPV1 expression in the lower thoracic and upper lumbar segments of spinal cord, DRG, and mesenteric arteries enhanced the prohypertensive effects of high salt intake, suggesting that TRPV1 channels in these sites protect against increased salt sensitivity, possibly via suppression of sympatho-excitatory responses. PMID- 21642953 TI - Structural basis of cargo recognition by the myosin-X MyTH4-FERM domain. AB - Myosin-X is an important unconventional myosin that is critical for cargo transportation to filopodia tips and is also utilized in spindle assembly by interacting with microtubules. We present a series of structural and biochemical studies of the myosin-X tail domain cassette, consisting of myosin tail homology 4 (MyTH4) and FERM domains in complex with its specific cargo, a netrin receptor DCC (deleted in colorectal cancer). The MyTH4 domain is folded into a helical VHS like structure and is associated with the FERM domain. We found an unexpected binding mode of the DCC peptide to the subdomain C groove of the FERM domain, which is distinct from previously reported beta-beta associations found in radixin-adhesion molecule complexes. We also revealed direct interactions between the MyTH4-FERM cassette and tubulin C-terminal acidic tails, and identified a positively charged patch of the MyTH4 domain, which is involved in tubulin binding. We demonstrated that both DCC and integrin bindings interfere with microtubule binding and that DCC binding interferes with integrin binding. Our results provide the molecular basis by which myosin-X facilitates alternative dual binding to cargos and microtubules. PMID- 21642954 TI - Regulated inactivation of the spindle assembly checkpoint without functional mitotic spindles. AB - The spindle assembly checkpoint (SAC) arrests mitosis until bipolar attachment of spindle microtubules to all chromosomes is accomplished. However, when spindle formation is prevented and the SAC cannot be satisfied, mammalian cells can eventually overcome the mitotic arrest while the checkpoint is still activated. We find that Aspergillus nidulans cells, which are unable to satisfy the SAC, inactivate the checkpoint after a defined period of mitotic arrest. Such SAC inactivation allows normal nuclear reassembly and mitotic exit without DNA segregation. We demonstrate that the mechanisms, which govern such SAC inactivation, require protein synthesis and can occur independently of inactivation of the major mitotic regulator Cdk1/Cyclin B or mitotic exit. Moreover, in the continued absence of spindle function cells transit multiple cell cycles in which the SAC is reactivated each mitosis before again being inactivated. Such cyclic activation and inactivation of the SAC suggests that it is subject to cell-cycle regulation that is independent of bipolar spindle function. PMID- 21642955 TI - Tra1 has specific regulatory roles, rather than global functions, within the SAGA co-activator complex. AB - The SAGA complex is a conserved, multifunctional co-activator that has broad roles in eukaryotic transcription. Previous studies suggested that Tra1, the largest SAGA component, is required either for SAGA assembly or for SAGA recruitment by DNA-bound transcriptional activators. In contrast to Saccharomyces cerevisiae and mouse, a tra1Delta mutant is viable in Schizosaccharomyces pombe, allowing us to test these issues in vivo. We find that, in a tra1Delta mutant, SAGA assembles and is recruited to some, but not all, promoters. Consistent with these findings, Tra1 regulates the expression of only a subset of SAGA-dependent genes. We previously reported that the SAGA subunits Gcn5 and Spt8 have opposing regulatory roles during S. pombe sexual differentiation. We show here that, like Gcn5, Tra1 represses this pathway, although by a distinct mechanism. Thus, our study reveals that Tra1 has specific regulatory roles, rather than global functions, within SAGA. PMID- 21642956 TI - An autism-associated point mutation in the neuroligin cytoplasmic tail selectively impairs AMPA receptor-mediated synaptic transmission in hippocampus. AB - Neuroligins are evolutionarily conserved postsynaptic cell-adhesion molecules that function, at least in part, by forming trans-synaptic complexes with presynaptic neurexins. Different neuroligin isoforms perform diverse functions and exhibit distinct intracellular localizations, but contain similar cytoplasmic sequences whose role remains largely unknown. Here, we analysed the effect of a single amino-acid substitution (R704C) that targets a conserved arginine residue in the cytoplasmic sequence of all neuroligins, and that was associated with autism in neuroligin-4. We introduced the R704C mutation into mouse neuroligin-3 by homologous recombination, and examined its effect on synapses in vitro and in vivo. Electrophysiological and morphological studies revealed that the neuroligin 3 R704C mutation did not significantly alter synapse formation, but dramatically impaired synapse function. Specifically, the R704C mutation caused a major and selective decrease in AMPA receptor-mediated synaptic transmission in pyramidal neurons of the hippocampus, without similarly changing NMDA or GABA receptor mediated synaptic transmission, and without detectably altering presynaptic neurotransmitter release. Our results suggest that the cytoplasmic tail of neuroligin-3 has a central role in synaptic transmission by modulating the recruitment of AMPA receptors to postsynaptic sites at excitatory synapses. PMID- 21642957 TI - Nek9 is a Plk1-activated kinase that controls early centrosome separation through Nek6/7 and Eg5. AB - The NIMA-family kinases Nek9/Nercc1, Nek6 and Nek7 form a signalling module required for mitotic spindle assembly. Nek9, the upstream kinase, is activated during prophase at centrosomes although the details of this have remained elusive. We now identify Plk1 as Nek9 direct activator and propose a two-step activation mechanism that involves Nek9 sequential phosphorylation by CDK1 and Plk1. Furthermore, we show that Plk1 controls prophase centrosome separation through the activation of Nek9 and ultimately the phosphorylation of the mitotic kinesin Eg5 at Ser1033, a Nek6/7 site that together with the CDK1 site Thr926 we establish contributes to the accumulation of Eg5 at centrosomes and is necessary for subsequent centrosome separation and timely mitosis. Our results provide a basis to understand signalling downstream of Plk1 and shed light on the role of Eg5, Plk1 and the NIMA-family kinases in the control of centrosome separation and normal mitotic progression. PMID- 21642958 TI - Distinct functional outputs of PTEN signalling are controlled by dynamic association with beta-arrestins. AB - The tumour suppressor PTEN (phosphatase and tensin deleted on chromosome 10) regulates major cellular functions via lipid phosphatase-dependent and independent mechanisms. Despite its fundamental pathophysiological importance, how PTEN's cellular activity is regulated has only been partially elucidated. We report that the scaffolding proteins beta-arrestins (beta-arrs) are important regulators of PTEN. Downstream of receptor-activated RhoA/ROCK signalling, beta arrs activate the lipid phosphatase activity of PTEN to negatively regulate Akt and cell proliferation. In contrast, following wound-induced RhoA activation, beta-arrs inhibit the lipid phosphatase-independent anti-migratory effects of PTEN. beta-arrs can thus differentially control distinct functional outputs of PTEN important for cell proliferation and migration. PMID- 21642959 TI - Dynein struts its stuff. PMID- 21642961 TI - A low mass for Mars from Jupiter's early gas-driven migration. AB - Jupiter and Saturn formed in a few million years (ref. 1) from a gas-dominated protoplanetary disk, and were susceptible to gas-driven migration of their orbits on timescales of only ~100,000 years (ref. 2). Hydrodynamic simulations show that these giant planets can undergo a two-stage, inward-then-outward, migration. The terrestrial planets finished accreting much later, and their characteristics, including Mars' small mass, are best reproduced by starting from a planetesimal disk with an outer edge at about one astronomical unit from the Sun (1 au is the Earth-Sun distance). Here we report simulations of the early Solar System that show how the inward migration of Jupiter to 1.5 au, and its subsequent outward migration, lead to a planetesimal disk truncated at 1 au; the terrestrial planets then form from this disk over the next 30-50 million years, with an Earth/Mars mass ratio consistent with observations. Scattering by Jupiter initially empties but then repopulates the asteroid belt, with inner-belt bodies originating between 1 and 3 au and outer-belt bodies originating between and beyond the giant planets. This explains the significant compositional differences across the asteroid belt. The key aspect missing from previous models of terrestrial planet formation is the substantial radial migration of the giant planets, which suggests that their behaviour is more similar to that inferred for extrasolar planets than previously thought. PMID- 21642962 TI - Whole-genome sequencing identifies recurrent mutations in chronic lymphocytic leukaemia. AB - Chronic lymphocytic leukaemia (CLL), the most frequent leukaemia in adults in Western countries, is a heterogeneous disease with variable clinical presentation and evolution. Two major molecular subtypes can be distinguished, characterized respectively by a high or low number of somatic hypermutations in the variable region of immunoglobulin genes. The molecular changes leading to the pathogenesis of the disease are still poorly understood. Here we performed whole-genome sequencing of four cases of CLL and identified 46 somatic mutations that potentially affect gene function. Further analysis of these mutations in 363 patients with CLL identified four genes that are recurrently mutated: notch 1 (NOTCH1), exportin 1 (XPO1), myeloid differentiation primary response gene 88 (MYD88) and kelch-like 6 (KLHL6). Mutations in MYD88 and KLHL6 are predominant in cases of CLL with mutated immunoglobulin genes, whereas NOTCH1 and XPO1 mutations are mainly detected in patients with unmutated immunoglobulins. The patterns of somatic mutation, supported by functional and clinical analyses, strongly indicate that the recurrent NOTCH1, MYD88 and XPO1 mutations are oncogenic changes that contribute to the clinical evolution of the disease. To our knowledge, this is the first comprehensive analysis of CLL combining whole-genome sequencing with clinical characteristics and clinical outcomes. It highlights the usefulness of this approach for the identification of clinically relevant mutations in cancer. PMID- 21642963 TI - Sharper low-power STED nanoscopy by time gating. AB - Applying pulsed excitation together with time-gated detection improves the fluorescence on-off contrast in continuous-wave stimulated emission depletion (CW STED) microscopy, thus revealing finer details in fixed and living cells using moderate light intensities. This method also enables super-resolution fluorescence correlation spectroscopy with CW-STED beams, as demonstrated by quantifying the dynamics of labeled lipid molecules in the plasma membrane of living cells. PMID- 21642964 TI - High-throughput behavioral analysis in C. elegans. AB - We designed a real-time computer vision system, the Multi-Worm Tracker (MWT), which can simultaneously quantify the behavior of dozens of Caenorhabditis elegans on a Petri plate at video rates. We examined three traditional behavioral paradigms using this system: spontaneous movement on food, where the behavior changes over tens of minutes; chemotaxis, where turning events must be detected accurately to determine strategy; and habituation of response to tap, where the response is stochastic and changes over time. In each case, manual analysis or automated single-worm tracking would be tedious and time-consuming, but the MWT system allowed rapid quantification of behavior with minimal human effort. Thus, this system will enable large-scale forward and reverse genetic screens for complex behaviors. PMID- 21642965 TI - Single-tube linear DNA amplification (LinDA) for robust ChIP-seq. AB - Genome-wide profiling of transcription factors based on massive parallel sequencing of immunoprecipitated chromatin (ChIP-seq) requires nanogram amounts of DNA. Here we describe a high-fidelity, single-tube linear DNA amplification method (LinDA) for ChIP-seq and reChIP-seq with picogram DNA amounts obtained from a few thousand cells. This amplification technology will facilitate global analyses of transcription-factor binding and chromatin with very small cell populations, such as stem or cancer-initiating cells. PMID- 21642966 TI - Near-infrared branding efficiently correlates light and electron microscopy. AB - The correlation of light and electron microscopy of complex tissues remains a major challenge. Here we report near-infrared branding (NIRB), which facilitates such correlation by using a pulsed, near-infrared laser to create defined fiducial marks in three dimensions in fixed tissue. As these marks are fluorescent and can be photo-oxidized to generate electron contrast, they can guide re-identification of previously imaged structures as small as dendritic spines by electron microscopy. PMID- 21642967 TI - Mechanism and function of synaptotagmin-mediated membrane apposition. AB - Synaptotagmin-1 is a Ca(2+) sensor that triggers synchronous neurotransmitter release. The first documented biochemical property of synaptotagmin-1 was its ability to aggregate membranes in response to Ca(2+). However, the mechanism and function of this process were poorly understood. Here we show that synaptotagmin 1-mediated vesicle aggregation is driven by trans interactions between synaptotagmin-1 molecules bound to different membranes. We found a strong correlation between the ability of Ca(2+)-bound synaptotagmin-1 to aggregate vesicles and to stimulate SNARE-mediated membrane fusion. Moreover, artificial aggregation of membranes-using non-synaptotagmin proteins-also efficiently promoted fusion of SNARE-bearing liposomes. Finally, using a modified fusion assay, we observed that synaptotagmin-1 drove the assembly of otherwise non fusogenic individual t-SNARE proteins into fusion-competent heterodimers, independently of aggregation. Thus, membrane aggregation and t-SNARE assembly appear to be two key aspects of fusion reactions that are regulated by Ca(2+) bound synaptotagmin-1 and catalyzed by SNAREs. PMID- 21642968 TI - Synaptotagmin-1 may be a distance regulator acting upstream of SNARE nucleation. AB - Synaptotagmin-1 triggers Ca(2+)-sensitive, rapid neurotransmitter release by promoting interactions between SNARE proteins on synaptic vesicles and the plasma membrane. How synaptotagmin-1 promotes this interaction is unclear, and the massive increase in membrane fusion efficiency of Ca(2+)-bound synaptotagmin-1 has not been reproduced in vitro. However, previous experiments have been performed at relatively high salt concentrations, screening potentially important electrostatic interactions. Using functional reconstitution in liposomes, we show here that at low ionic strength SNARE-mediated membrane fusion becomes strictly dependent on both Ca(2+) and synaptotagmin-1. Under these conditions, synaptotagmin-1 functions as a distance regulator that tethers the liposomes too far from the plasma membrane for SNARE nucleation in the absence of Ca(2+), but while bringing the liposomes close enough for membrane fusion in the presence of Ca(2+). These results may explain how the relatively weak electrostatic interactions between synaptotagmin-1 and membranes substantially accelerate fusion. PMID- 21642969 TI - KAP-1 phosphorylation regulates CHD3 nucleosome remodeling during the DNA double strand break response. AB - KAP-1 poses a substantial barrier to DNA double-strand break (DSB) repair within heterochromatin that is alleviated by ATM-dependent KAP-1 phosphorylation (pKAP 1). Here we address the mechanistic consequences of pKAP-1 that promote heterochromatic DSB repair and chromatin relaxation. KAP-1 function involves autoSUMOylation and recruitment of nucleosome deacetylation, methylation and remodeling activities. Although heterochromatin acetylation or methylation changes were not detected, radiation-induced pKAP-1 dispersed the nucleosome remodeler CHD3 from DSBs and triggered concomitant chromatin relaxation; pKAP-1 loss reversed these effects. Depletion or inactivation of CHD3, or ablation of its interaction with KAP-1(SUMO1), bypassed pKAP-1's role in repair. Though KAP-1 SUMOylation was unaffected after irradiation, CHD3 dissociated from KAP-1(SUMO1) in a pKAP-1-dependent manner. We demonstrate that KAP-1(Ser824) phosphorylation generates a motif that directly perturbs interactions between CHD3's SUMO interacting motif and SUMO1, dispersing CHD3 from heterochromatin DSBs and enabling repair. PMID- 21642970 TI - Structure-function studies of FMRP RGG peptide recognition of an RNA duplex quadruplex junction. AB - We have determined the solution structure of the complex between an arginine glycine-rich RGG peptide from the human fragile X mental retardation protein (FMRP) and an in vitro-selected guanine-rich (G-rich) sc1 RNA. The bound RNA forms a newly discovered G-quadruplex separated from the flanking duplex stem by a mixed junctional tetrad. The RGG peptide is positioned along the major groove of the RNA duplex, with the G-quadruplex forcing a sharp turn of R(10)GGGGR(15) at the duplex-quadruplex junction. Arg10 and Arg15 form cross-strand specificity determining intermolecular hydrogen bonds with the major-groove edges of guanines of adjacent Watson-Crick G*C pairs. Filter-binding assays on RNA and peptide mutations identify and validate contributions of peptide-RNA intermolecular contacts and shape complementarity to molecular recognition. These findings on FMRP RGG domain recognition by a combination of G-quadruplex and surrounding RNA sequences have implications for the recognition of other genomic G-rich RNAs. PMID- 21642971 TI - Crystal structure of the trithorax group protein ASH2L reveals a forkhead-like DNA binding domain. AB - Absent, small or homeotic discs-like 2 (ASH2L) is a trithorax group (TrxG) protein and a regulatory subunit of the SET1 family of lysine methyltransferases. Here we report that ASH2L binds DNA using a forkhead-like helix-wing-helix (HWH) domain. In vivo, the ASH2L HWH domain is required for binding to the beta-globin locus control region, histone H3 Lys4 (H3K4) trimethylation and maximal expression of the beta-globin gene (Hbb-1), validating the functional importance of the ASH2L DNA binding domain. PMID- 21642973 TI - Neuronal basis of sequential foraging decisions in a patchy environment. AB - Deciding when to leave a depleting resource to exploit another is a fundamental problem for all decision makers. The neuronal mechanisms mediating patch-leaving decisions remain unknown. We found that neurons in primate (Macaca mulatta) dorsal anterior cingulate cortex, an area that is linked to reward monitoring and executive control, encode a decision variable signaling the relative value of leaving a depleting resource for a new one. Neurons fired during each sequential decision to stay in a patch and, for each travel time, these responses reached a fixed threshold for patch-leaving. Longer travel times reduced the gain of neural responses for choosing to stay in a patch and increased the firing rate threshold mandating patch-leaving. These modulations more closely matched behavioral decisions than any single task variable. These findings portend an understanding of the neural basis of foraging decisions and endorse the unification of theoretical and experimental work in ecology and neuroscience. PMID- 21642972 TI - UNC119 is required for G protein trafficking in sensory neurons. AB - UNC119 is widely expressed among vertebrates and other phyla. We found that UNC119 recognized the acylated N terminus of the rod photoreceptor transducin alpha (Talpha) subunit and Caenorhabditis elegans G proteins ODR-3 and GPA-13. The crystal structure of human UNC119 at 1.95-A resolution revealed an immunoglobulin-like beta-sandwich fold. Pulldowns and isothermal titration calorimetry revealed a tight interaction between UNC119 and acylated Galpha peptides. The structure of co-crystals of UNC119 with an acylated Talpha N terminal peptide at 2.0 A revealed that the lipid chain is buried deeply into UNC119's hydrophobic cavity. UNC119 bound Talpha-GTP, inhibiting its GTPase activity, thereby providing a stable UNC119-Talpha-GTP complex capable of diffusing from the inner segment back to the outer segment after light-induced translocation. UNC119 deletion in both mouse and C. elegans led to G protein mislocalization. Thus, UNC119 is a Galpha subunit cofactor essential for G protein trafficking in sensory cilia. PMID- 21642974 TI - Light acts through melanopsin to alter retinal waves and segregation of retinogeniculate afferents. AB - Waves of correlated activity sweeping across the early postnatal mouse retina promote the segregation and refinement of retinofugal projections. This process has been thought to be spontaneous and unaffected by visual experience. We found, however, that light prolongs spiking during the waves and enhances the segregation of retinogeniculate afferents, and that it did so by activating melanopsin-expressing, intrinsically photosensitive retinal ganglion cells. PMID- 21642975 TI - High-fat feeding promotes obesity via insulin receptor/PI3K-dependent inhibition of SF-1 VMH neurons. AB - Steroidogenic factor 1 (SF-1)-expressing neurons of the ventromedial hypothalamus (VMH) control energy homeostasis, but the role of insulin action in these cells remains undefined. We show that insulin activates phosphatidylinositol-3-OH kinase (PI3K) signaling in SF-1 neurons and reduces firing frequency in these cells through activation of K(ATP) channels. These effects were abrogated in mice with insulin receptor deficiency restricted to SF-1 neurons (SF-1(DeltaIR) mice). Whereas body weight and glucose homeostasis remained the same in SF-1(DeltaIR) mice as in controls under a normal chow diet, they were protected from diet induced leptin resistance, weight gain, adiposity and impaired glucose tolerance. High-fat feeding activated PI3K signaling in SF-1 neurons of control mice, and this response was attenuated in the VMH of SF-1(DeltaIR) mice. Mimicking diet induced overactivation of PI3K signaling by disruption of the phosphatidylinositol-3,4,5-trisphosphate phosphatase PTEN led to increased body weight and hyperphagia under a normal chow diet. Collectively, our experiments reveal that high-fat diet-induced, insulin-dependent PI3K activation in VMH neurons contributes to obesity development. PMID- 21642976 TI - Cardinal rules: visual orientation perception reflects knowledge of environmental statistics. AB - Humans are good at performing visual tasks, but experimental measurements have revealed substantial biases in the perception of basic visual attributes. An appealing hypothesis is that these biases arise through a process of statistical inference, in which information from noisy measurements is fused with a probabilistic model of the environment. However, such inference is optimal only if the observer's internal model matches the environment. We found this to be the case. We measured performance in an orientation-estimation task and found that orientation judgments were more accurate at cardinal (horizontal and vertical) orientations. Judgments made under conditions of uncertainty were strongly biased toward cardinal orientations. We estimated observers' internal models for orientation and found that they matched the local orientation distribution measured in photographs. In addition, we determined how a neural population could embed probabilistic information responsible for such biases. PMID- 21642977 TI - Simultaneous two-photon imaging of oxygen and blood flow in deep cerebral vessels. AB - Uncovering principles that regulate energy metabolism in the brain requires mapping of partial pressure of oxygen (PO(2)) and blood flow with high spatial and temporal resolution. Using two-photon phosphorescence lifetime microscopy (2PLM) and the oxygen probe PtP-C343, we show that PO(2) can be accurately measured in the brain at depths up to 300 MUm with micron-scale resolution. In addition, 2PLM allowed simultaneous measurements of blood flow and of PO(2) in capillaries with less than one-second temporal resolution. Using this approach, we detected erythrocyte-associated transients (EATs) in oxygen in the rat olfactory bulb and showed the existence of diffusion-based arterio-venous shunts. Sensory stimulation evoked functional hyperemia, accompanied by an increase in PO(2) in capillaries and by a biphasic PO(2) response in the neuropil, consisting of an 'initial dip' and a rebound. 2PLM of PO(2) opens new avenues for studies of brain metabolism and blood flow regulation. PMID- 21642978 TI - Uncoupling the mechanisms of obesity and hypertension by targeting hypothalamic IKK-beta and NF-kappaB. AB - Obesity-related hypertension has become an epidemic health problem and a major risk factor for the development of cardiovascular disease (CVD). Recent research on the pathophysiology of obesity has implicated a role for the hypothalamus in the pathogenesis of this condition. However, it remains unknown whether the often seen coupling of hypertension with obesity can also be explained by hypothalamic dysfunction, despite the emerging appreciation that many forms of hypertension are neurogenic in origin. Our studies here revealed that acute activation of the proinflammatory protein nuclear factor kappaB (NF-kappaB) and its upstream activator IkappaB kinase-beta (IKK-beta, encoded by Ikbkb) in the mediobasal hypothalamus rapidly elevated blood pressure in mice independently of obesity. This form of hypothalamic inflammation-induced hypertension involved the sympathetic upregulation of hemodynamics and was reversed by sympathetic suppression. Loss-of-function studies further showed that NF-kappaB inhibition in the mediobasal hypothalamus counteracted obesity-related hypertension in a manner that was dissociable from changes in body weight. In addition, we found that pro opiomelanocortin (POMC) neurons were crucial for the hypertensive effects of the activation of hypothalamic IKK-beta and NF-kappaB, which underlie obesity-related hypertension. In conclusion, obesity-associated activation of IKK-beta and NF kappaB in the mediobasal hypothalamus--particularly in the hypothalamic POMC neurons--is a primary pathogenic link between obesity and hypertension. Breaking this pathogenic link may represent an avenue for controlling obesity-related hypertension and CVD without requiring obesity control. PMID- 21642979 TI - Suppression of inflammatory and neuropathic pain by uncoupling CRMP-2 from the presynaptic Ca2+ channel complex. AB - The use of N-type voltage-gated calcium channel (CaV2.2) blockers to treat pain is limited by many physiological side effects. Here we report that inflammatory and neuropathic hypersensitivity can be suppressed by inhibiting the binding of collapsin response mediator protein 2 (CRMP-2) to CaV2.2 and thereby reducing channel function. A peptide of CRMP-2 fused to the HIV transactivator of transcription (TAT) protein (TAT-CBD3) decreased neuropeptide release from sensory neurons and excitatory synaptic transmission in dorsal horn neurons, reduced meningeal blood flow, reduced nocifensive behavior induced by formalin injection or corneal capsaicin application and reversed neuropathic hypersensitivity produced by an antiretroviral drug. TAT-CBD3 was mildly anxiolytic without affecting memory retrieval, sensorimotor function or depression. At doses tenfold higher than that required to reduce hypersensitivity in vivo, TAT-CBD3 caused a transient episode of tail kinking and body contortion. By preventing CRMP-2-mediated enhancement of CaV2.2 function, TAT-CBD3 alleviated inflammatory and neuropathic hypersensitivity, an approach that may prove useful in managing chronic pain. PMID- 21642980 TI - A systematic RNAi synthetic interaction screen reveals a link between p53 and snoRNP assembly. AB - TP53 (tumour protein 53) is one of the most frequently mutated genes in human cancer and its role during cellular transformation has been studied extensively. However, the homeostatic functions of p53 are less well understood. Here, we explore the molecular dependency network of TP53 through an RNAi-mediated synthetic interaction screen employing two HCT116 isogenic cell lines and a genome-scale endoribonuclease-prepared short interfering RNA library. We identify a variety of TP53 synthetic interactions unmasking the complex connections of p53 to cellular physiology and growth control. Molecular dissection of the TP53 synthetic interaction with UNRIP indicates an enhanced dependency of TP53 negative cells on small nucleolar ribonucleoprotein (snoRNP) assembly. This dependency is mediated by the snoRNP chaperone gene NOLC1 (also known as NOPP140), which we identify as a physiological p53 target gene. This unanticipated function of TP53 in snoRNP assembly highlights the potential of RNAi-mediated synthetic interaction screens to dissect molecular pathways of tumour suppressor genes. PMID- 21642981 TI - A direct role for Met endocytosis in tumorigenesis. AB - Compartmentalization of signals generated by receptor tyrosine kinase (RTK) endocytosis has emerged as a major determinant of various cell functions. Here, using tumour-associated Met-activating mutations, we demonstrate a direct link between endocytosis and tumorigenicity. Met mutants exhibit increased endocytosis/recycling activity and decreased levels of degradation, leading to accumulation on endosomes, activation of the GTPase Rac1, loss of actin stress fibres and increased levels of cell migration. Blocking endocytosis inhibited mutants' anchorage-independent growth, in vivo tumorigenesis and metastasis while maintaining their activation. One mutant resistant to inhibition by a Met specific tyrosine kinase inhibitor was sensitive to endocytosis inhibition. Thus, oncogenicity of Met mutants results not only from activation but also from their altered endocytic trafficking, indicating that endosomal signalling may be a crucial mechanism regulating RTK-dependent tumorigenesis. PMID- 21642982 TI - Intraflagellar transport delivers tubulin isotypes to sensory cilium middle and distal segments. AB - Sensory cilia are assembled and maintained by kinesin-2-dependent intraflagellar transport (IFT). We investigated whether two Caenorhabditis elegans alpha- and beta-tubulin isotypes, identified through mutants that lack their cilium distal segments, are delivered to their assembly sites by IFT. Mutations in conserved residues in both tubulins destabilize distal singlet microtubules. One isotype, TBB-4, assembles into microtubules at the tips of the axoneme core and distal segments, where the microtubule tip tracker EB1 is found, and localizes all along the cilium, whereas the other, TBA-5, concentrates in distal singlets. IFT assays, fluorescence recovery after photobleaching analysis and modelling indicate that the continual transport of sub-stoichiometric numbers of these tubulin subunits by the IFT machinery can maintain sensory cilia at their steady state length. PMID- 21642983 TI - Giant anharmonic phonon scattering in PbTe. AB - Understanding the microscopic processes affecting the bulk thermal conductivity is crucial to develop more efficient thermoelectric materials. PbTe is currently one of the leading thermoelectric materials, largely thanks to its low thermal conductivity. However, the origin of this low thermal conductivity in a simple rocksalt structure has so far been elusive. Using a combination of inelastic neutron scattering measurements and first-principles computations of the phonons, we identify a strong anharmonic coupling between the ferroelectric transverse optic mode and the longitudinal acoustic modes in PbTe. This interaction extends over a large portion of reciprocal space, and directly affects the heat-carrying longitudinal acoustic phonons. The longitudinal acoustic-transverse optic anharmonic coupling is likely to play a central role in explaining the low thermal conductivity of PbTe. The present results provide a microscopic picture of why many good thermoelectric materials are found near a lattice instability of the ferroelectric type. PMID- 21642984 TI - Large-area flexible 3D optical negative index metamaterial formed by nanotransfer printing. AB - Negative-index metamaterials (NIMs) are engineered structures with optical properties that cannot be obtained in naturally occurring materials. Recent work has demonstrated that focused ion beam and layer-by-layer electron-beam lithography can be used to pattern the necessary nanoscale features over small areas (hundreds of um(2)) for metamaterials with three-dimensional layouts and interesting characteristics, including negative-index behaviour in the optical regime. A key challenge is in the fabrication of such three-dimensional NIMs with sizes and at throughputs necessary for many realistic applications (including lenses, resonators and other photonic components). We report a simple printing approach capable of forming large-area, high-quality NIMs with three-dimensional, multilayer formats. Here, a silicon wafer with deep, nanoscale patterns of surface relief serves as a reusable stamp. Blanket deposition of alternating layers of silver and magnesium fluoride onto such a stamp represents a process for 'inking' it with thick, multilayer assemblies. Transfer printing this ink material onto rigid or flexible substrates completes the fabrication in a high throughput manner. Experimental measurements and simulation results show that macroscale, three-dimensional NIMs (>75 cm(2)) nano-manufactured in this way exhibit a strong, negative index of refraction in the near-infrared spectral range, with excellent figures of merit. PMID- 21642985 TI - Reveromycin A biosynthesis uses RevG and RevJ for stereospecific spiroacetal formation. AB - Spiroacetal compounds are ubiquitous in nature, and their stereospecific structures are responsible for diverse pharmaceutical activities. Elucidation of the biosynthetic mechanisms that are involved in spiroacetal formation will open the door to efficient generation of stereospecific structures that are otherwise hard to synthesize chemically. However, the biosynthesis of these compounds is poorly understood, owing to difficulties in identifying the responsible enzymes and analyzing unstable intermediates. Here we comprehensively describe the spiroacetal formation involved in the biosynthesis of reveromycin A, which inhibits bone resorption and bone metastases of tumor cells by inducing apoptosis in osteoclasts. We performed gene disruption, systematic metabolite analysis, feeding of labeled precursors and conversion studies with recombinant enzymes. We identified two key enzymes, dihydroxy ketone synthase and spiroacetal synthase, and showed in vitro reconstruction of the stereospecific spiroacetal structure from a stable acyclic precursor. Our findings provide insights into the creation of a variety of biologically active spiroacetal compounds for drug leads. PMID- 21642986 TI - Pre-existing clusters of the adaptor Lat do not participate in early T cell signaling events. AB - Engaged T cell antigen receptors (TCRs) initiate signaling through the adaptor protein Lat. In quiescent T cells, Lat is segregated into clusters on the cell surface, which raises the question of how TCR triggering initiates signaling. Using super-resolution fluorescence microscopy, we found that pre-existing Lat domains were neither phosphorylated nor laterally transported to TCR activation sites, which suggested that these clusters do not participate in TCR signaling. Instead, TCR activation resulted in the recruitment and phosphorylation of Lat from subsynaptic vesicles. Studies of Lat mutants confirmed that recruitment preceded and was essential for phosphorylation and that both processes were independent of surface clustering of Lat. Our data suggest that TCR ligation preconditions the membrane for vesicle recruitment and bulk activation of the Lat signaling network. PMID- 21642987 TI - IFIT1 is an antiviral protein that recognizes 5'-triphosphate RNA. AB - Antiviral innate immunity relies on the recognition of microbial structures. One such structure is viral RNA that carries a triphosphate group on its 5' terminus (PPP-RNA). By an affinity proteomics approach with PPP-RNA as the 'bait', we found that the antiviral protein IFIT1 (interferon-induced protein with tetratricopeptide repeats 1) mediated binding of a larger protein complex containing other IFIT family members. IFIT1 bound PPP-RNA with nanomolar affinity and required the arginine at position 187 in a highly charged carboxy-terminal groove of the protein. In the absence of IFIT1, the growth and pathogenicity of viruses containing PPP-RNA was much greater. In contrast, IFIT proteins were dispensable for the clearance of pathogens that did not generate PPP-RNA. On the basis of this specificity and the great abundance of IFIT proteins after infection, we propose that the IFIT complex antagonizes viruses by sequestering specific viral nucleic acids. PMID- 21642988 TI - The sphingosine 1-phosphate receptor S1P2 maintains the homeostasis of germinal center B cells and promotes niche confinement. AB - Mice deficient in sphingosine 1-phosphate receptor type 2 (S1P(2)) develop diffuse large B cell lymphoma. However, the role of S1P(2) in normal germinal center (GC) physiology is unknown. Here we show that S1P(2)-deficient GC B cells outgrew their wild-type counterparts in chronically established GCs. We found that antagonism of the kinase Akt mediated by S1P(2) and its downstream mediators Galpha(12), Galpha(13) and p115RhoGEF regulated cell viability and was required for growth control in chronically proliferating GCs. Moreover, S1P(2) inhibited GC B cell responses to follicular chemoattractants and helped confine cells to the GC. In addition, S1P(2) overexpression promoted the centering of activated B cells in the follicle. We suggest that by inhibiting Akt activation and migration, S1P(2) helps restrict GC B cell survival and localization to an S1P low niche at the follicle center. PMID- 21642989 TI - Arabidopsis REF6 is a histone H3 lysine 27 demethylase. AB - Polycomb group (PcG)-mediated histone H3 lysine 27 trimethylation (H3K27me3) has a key role in gene repression and developmental regulation. There is evidence that H3K27me3 is actively removed in plants, but it is not known how this occurs. Here we show that RELATIVE OF EARLY FLOWERING 6 (REF6), also known as Jumonji domain-containing protein 12 (JMJ12), specifically demethylates H3K27me3 and H3K27me2, whereas its metazoan counterparts, the KDM4 proteins, are H3K9 and H3K36 demethylases. Plants overexpressing REF6 resembled mutants defective in H3K27me3-mediated gene silencing. Genetic interaction tests indicated that REF6 acts downstream of H3K27me3 methyltransferases. Mutations in REF6 caused ectopic and increased H3K27me3 level and decreased mRNA expression of hundreds of genes involved in regulating developmental patterning and responses to various stimuli. Our work shows that plants and metazoans use conserved mechanisms to regulate H3K27me3 dynamics but use distinct subfamilies of enzymes. PMID- 21642990 TI - A cooperative microRNA-tumor suppressor gene network in acute T-cell lymphoblastic leukemia (T-ALL). AB - The importance of individual microRNAs (miRNAs) has been established in specific cancers. However, a comprehensive analysis of the contribution of miRNAs to the pathogenesis of any specific cancer is lacking. Here we show that in T-cell acute lymphoblastic leukemia (T-ALL), a small set of miRNAs is responsible for the cooperative suppression of several tumor suppressor genes. Cross-comparison of miRNA expression profiles in human T-ALL with the results of an unbiased miRNA library screen allowed us to identify five miRNAs (miR-19b, miR-20a, miR-26a, miR 92 and miR-223) that are capable of promoting T-ALL development in a mouse model and which account for the majority of miRNA expression in human T-ALL. Moreover, these miRNAs produce overlapping and cooperative effects on tumor suppressor genes implicated in the pathogenesis of T-ALL, including IKAROS (also known as IKZF1), PTEN, BIM, PHF6, NF1 and FBXW7. Thus, a comprehensive and unbiased analysis of miRNA action in T-ALL reveals a striking pattern of miRNA-tumor suppressor gene interactions in this cancer. PMID- 21642991 TI - The nuclear deubiquitinase BAP1 is commonly inactivated by somatic mutations and 3p21.1 losses in malignant pleural mesothelioma. AB - Malignant pleural mesotheliomas (MPMs) often show CDKN2A and NF2 inactivation, but other highly recurrent mutations have not been described. To identify additional driver genes, we used an integrated genomic analysis of 53 MPM tumor samples to guide a focused sequencing effort that uncovered somatic inactivating mutations in BAP1 in 23% of MPMs. The BAP1 nuclear deubiquitinase is known to target histones (together with ASXL1 as a Polycomb repressor subunit) and the HCF1 transcriptional co-factor, and we show that BAP1 knockdown in MPM cell lines affects E2F and Polycomb target genes. These findings implicate transcriptional deregulation in the pathogenesis of MPM. PMID- 21642992 TI - Extensive and coordinated transcription of noncoding RNAs within cell-cycle promoters. AB - Transcription of long noncoding RNAs (lncRNAs) within gene regulatory elements can modulate gene activity in response to external stimuli, but the scope and functions of such activity are not known. Here we use an ultrahigh-density array that tiles the promoters of 56 cell-cycle genes to interrogate 108 samples representing diverse perturbations. We identify 216 transcribed regions that encode putative lncRNAs, many with RT-PCR-validated periodic expression during the cell cycle, show altered expression in human cancers and are regulated in expression by specific oncogenic stimuli, stem cell differentiation or DNA damage. DNA damage induces five lncRNAs from the CDKN1A promoter, and one such lncRNA, named PANDA, is induced in a p53-dependent manner. PANDA interacts with the transcription factor NF-YA to limit expression of pro-apoptotic genes; PANDA depletion markedly sensitized human fibroblasts to apoptosis by doxorubicin. These findings suggest potentially widespread roles for promoter lncRNAs in cell growth control. PMID- 21642994 TI - Analysis of vascular function using the cardio-ankle vascular index (CAVI). PMID- 21642993 TI - Genome-wide association study identifies three new susceptibility loci for esophageal squamous-cell carcinoma in Chinese populations. AB - Esophageal squamous-cell carcinoma (ESCC) is one of the most prevalent cancers worldwide and occurs at a relatively high frequency in China. To identify genetic susceptibility loci for ESCC, we conducted a genome-wide association study on 2,031 individuals with ESCC (cases) and 2,044 controls of Chinese descent using 666,141 autosomal SNPs. We evaluated promising associations in an additional 6,276 cases and 6,165 controls of Chinese descent from different areas of China. We identified seven susceptibility loci on chromosomes 5q11, 6p21, 10q23, 12q24 and 21q22 (ranging from P = 7.48 * 10(-12) to P = 2.44 * 10(-31)); among these loci, 5q11, 6p21 and 21q22 were newly identified. Three variants in high linkage disequilibrium on 12q24 confer their risks to ESCC in a gene-lifestyle interaction manner, with more pronounced risk enhancement seen in tobacco and alcohol users. Furthermore, the identified variants had a cumulative association with ESCC risk (P(trend) = 7.92 * 10(-56)). These findings highlight the involvement of multiple genetic loci and gene-environment interaction in the development of esophageal cancer. PMID- 21642995 TI - Generation of fertile sperm in a culture dish: clinical implications. PMID- 21642996 TI - What is appropriate neoadjuvant/adjuvant androgen deprivation for high risk/locally advanced prostate cancer? PMID- 21642997 TI - Andrology in China: current status and 10 years' progress. AB - Andrology has a long history in traditional Chinese medicine. There are records of male sexual health, male sexual dysfunction and male infertility from over thousands of years ago. Modern andrology in China had a late start, with the Chinese Andrology Association founded in 1995. Within last decade, andrology in China has grown rapidly. In this review article, we summarized the progress of andrology in last 10 years and outlined the current status of Chinese andrology with a special focus on progress in male erectile dysfunction, prostate diseases, male infertility and male hormonal contraception. PMID- 21642998 TI - The effects and prospects of the integration of traditional Chinese medicine and Western medicine on andrology in China. AB - Andrology has a long history in traditional Chinese medicine (TCM) discussions concerning andropathies, and documentation of relevant therapeutic methods abound in the ancient literature on TCM. Integrated treatment combining TCM and Western medicine has seen both broad and in-depth development, with formidable status in the field of modern andrology in China. This article attempts to demonstrate the unique advantage of integrated treatment in the therapy of andropathies through a review of the ancient literature on andrology in the field of TCM and on the integrative treatment of prostatic diseases, sexual dysfunction, male infertility and late-onset hypogonadism. There is a need for the advancement of a medical theory that integrates TCM and Western medicine practices to create a new therapeutic system with standardized therapeutic and evaluative protocols for diseases involving male sexual health. PMID- 21642999 TI - Differences in phenotype and gene expression of prostate stromal cells from patients of varying ages and their influence on tumour formation by prostate epithelial cells. AB - Prostate cancer (PCa) is an age-related disease, and the stromal microenvironment plays an important role in prostatic malignant progression. However, the differences in prostate stromal cells present in young and old tissue are still obscure. We established primary cultured stromal cells from normal prostatic peripheral zone (PZ) of donors of varying ages and found that cultured stromal cells from old donors (PZ-old) were more enlarged and polygonal than those from young donors (PZ-young). Furthermore, based on immunocytochemical and ultrastructural analysis, the components of stromal cells changed from a majority of fibroblasts to a mixture of fibroblasts and myofibroblasts with increasing donor age. Using a three-dimensional in vitro culture system, we found that PZ old stromal cells could enhance the proliferation, migration and invasion of cocultured benign BPH-1 and PC-3 cells. Using an in vivo tissue recombination system, we also found that PZ-old stromal cells are more effective than PZ-young cells in promoting tumour formation by BPH-1 cells of high passage (>100) and PC 3 cells. To probe the possible mechanism of these effects, we performed cDNA microarray analysis and profiled 509 upregulated genes and 188 downregulated genes in PZ-old cells. Among the changed genes, we found genes coding for a subset of paracrine factors that are capable of influencing adjacent epithelial cells; these include hepatocyte growth factor (HGF), fibroblast growth factor 5 (FGF5), insulin-like growth factor 2 (IGF2), insulin-like growth factor-binding protein 4 (IGFBP4), IGFBP5 and matrix metallopeptidase 1 (MMP1). Changes in the expression of these genes were further confirmed by quantitative real-time polymerase chain reaction (PCR), Western blotting and enzyme-linked immunosorbent assays. Overall, our findings indicate that stromal cells from prostate PZ of old donors are more active than similar cells from young donors in promoting the malignant process of adjacent epithelial cells. This finding hints at a new potential strategy for the prevention of PCa. PMID- 21643000 TI - Sexual medicine disparities between Asia and North America: commentary on male sexual dysfunction in Asia. PMID- 21643001 TI - Male sexual dysfunction in Asia. AB - Sex has always been a taboo subject in Asian society. However, over the past few years, awareness in the field of men's sexual health has improved, and interest in sexual health research has recently increased. The epidemiology and prevalence of erectile dysfunction, hypogonadism and premature ejaculation in Asia are similar in the West. However, several issues are specific to Asian males, including culture and beliefs, awareness, compliance and the availability of traditional/complementary medicine. In Asia, sexual medicine is still in its infancy, and a concerted effort from the government, relevant societies, physicians and the media is required to propel sexual medicine to the forefront of health care. PMID- 21643002 TI - The skeleton gets a (reproductive) life. PMID- 21643003 TI - The development and functions of CD4(+) T cells expressing a transgenic TCR specific for an MHC-I-restricted tumor antigenic epitope. AB - It has been reported that the ratio of CD4(+) to CD8(+) T cells has no bias in a few class I major histocompatibility complex (MHC-I)-restricted T-cell receptor (TCR)-transgenic mice specific for alloantigens or autoantigens, in which most CD4(+) T cells express an MHC-I-restricted TCR. In this study, we further showed that more than 50% of CD4(+) T cells in MHC-I-restricted P1A tumor antigen specific TCR (P1ATCR)-transgenic mice could specifically bind to MHC-I/P1A peptide complex. P1A peptide could stimulate the transgenic CD4(+) T cells to proliferate and secrete both type 1 helper T cell and type 2 helper T cell cytokines. The activated CD4(+) T cells also showed cytotoxicity against P1A expressing tumor cells. The analysis of TCR alpha-chains showed that these CD4(+) T cells were selected by co-expressing endogenous TCRs. Our results show that CD4(+) T cells from P1ATCR transgenic mice co-expressed an MHC-I-restricted transgenic TCR and another rearranged endogenous TCRs, both of which were functional. PMID- 21643004 TI - Establishment of platform for screening insulin-like growth factor-1 receptor inhibitors and evaluation of novel inhibitors. AB - AIM: The insulin-like growth factor-1 receptor (IGF1R) is over-expressed in a wide variety of tumors and contributes to tumor cell proliferation, metastasis and drug resistance. The aim of this study was to establish a sensitive screening platform to identify novel IGF1R inhibitors. METHODS: The catalytic domain of IGF1R was expressed using the Bac-to-Bac baculovirus expression system. The screening platform for IGF1R inhibitors was established based on ELISA. The binding profile of IGF1R with the inhibitors was predicted with molecular docking and then subjected to the surface plasmon resonance (SPR) approach. The growth inhibition of cancer cells by the inhibitors was assessed with MTT assay. Apoptosis was analyzed using flow cytometry and Western blotting. RESULTS: A naturally occurring small molecule compound hematoxylin was identified as the most potent inhibitor (IC50 value=1.8+/-0.1 MUmol/L) within a library of more than 200 compounds tested. Molecular simulation predicted the possible binding mode of hematoxylin with IGF1R. An SPR assay further confirmed that hematoxylin bound directly to IGF1R with high binding affinity (Kd=4.2 * 10-6 mol/L). In HL 60 cancer cells, hematoxylin inactivated the phosphorylation of IGF1R and downstream signaling and therefore suppressed cell proliferation. Mechanistic studies revealed that hematoxylin induced apoptosis in HL-60 cells via both extrinsic and intrinsic pathways. CONCLUSION: A simple, sensitive ELISA-based screening platform for identifying IGF1R inhibitors was established. Hematoxylin was identified as a promising IGF1R inhibitor with effective antitumor activity that deserves further investigation. PMID- 21643005 TI - A novel positive feedback loop involving FASN/p-ERK1/2/5-LOX/LTB4/FASN sustains high growth of breast cancer cells. AB - AIM: To investigate the endogenous signaling pathways associated with high proliferation potential of breast cancer cells. METHODS: Breast cancer cell lines LM-MCF-7 and MCF-7 with high and low proliferation capability were used. The promoter activity of fatty acid synthase (FASN) was examined using luciferase reporter gene assay. The expression level of FASN mRNA was measured using RT-PCR and real time PCR, respectively. The level of leukotriene B4 (LTB4) was determined with ELISA. The expression levels of 5-lipoxygenase (5-LOX) was analyzed using RT-PCR and Western blot, respectively. 5-Bromo-20-deoxyuridine (BrdU) incorporation assay was used to study the proliferation of LM-MCF-7 and MCF-7 cells. RESULTS: The promoter activity of FASN was significantly higher in LM-MCF-7 cells than MCF-7 cells. Treatment of LM-MCF-7 cells with ERK1/2 inhibitor PD98059 (30-50 MUmol/L) or LOX inhibitor NDGA (25 MUmol/L) abolished the activation of FASN. Moreover, treatment of LM-MCF-7 cells with the specific 5 LOX inhibitor MK-886 (20-40 MUmol/L) or 5-LOX siRNA (50-100 nmol/L) decreased the promoter activity of FASN. The level of LTB4, the final metabolite produced by 5 LOX, was significantly higher in LM-MCF-7 cells than MCF-7 cells. Administration of exogenous LTB4 (1-10 nmol/L) was able to stimulate the promoter activity of FASN in MCF-7 cells. Treatment of LM-MCF-7 cells with the FASN inhibitor cerulenin (10 MUmol/L) reduced all the levels of p-ERK1/2, 5-LOX, and LTB4. Treatment of LM-MCF-7 cells with cerulenin, PD98059, or MK-886 abolished the proliferation. Administration of exogenous LTB4 (10 nmol/L) significantly increased BrdU incorporation in MCF-7 cells. CONCLUSION: THESE results suggest a novel positive feedback loop involving FASN/p-ERK1/2/5-LOX/LTB4/FASN contributes to the sustaining growth of breast cancer LM-MCF-7 cells. PMID- 21643006 TI - Anthracyclines disrupt telomere maintenance by telomerase through inducing PinX1 ubiquitination and degradation. AB - Telomere maintenance is essential for cancer growth. Induction of telomere dysfunction, for example, by inhibition of telomeric proteins or telomerase, has been shown to strongly enhance cancer cells' sensitivity to chemotherapies. However, it is not clear whether modulations of telomere maintenance constitute cancer cellular responses to chemotherapies. Furthermore, the manner in which anti-cancer drugs affect telomere function remains unknown. In this study, we show that anthracyclines, a class of anti-cancer drugs widely used in clinical cancer treatments, have an active role in triggering telomere dysfunction specifically in telomerase-positive cancer cells. Anthracyclines interrupt telomere maintenance by telomerase through the downregulation of PinX1, a protein factor responsible for targeting telomerase onto telomeres, thereby inhibiting telomerase association with telomeres. We further demonstrate that anthracyclines downregulate PinX1 by inducing this protein degradation through the ubiquitin proteasome-dependent pathway. Our data not only reveal a novel action for anthracyclines as telomerase functional inhibitors but also provide a clue for the development of novel anti-cancer drugs based on telomerase/telomere targeting, which is actively investigated by many current studies. PMID- 21643007 TI - Conserved features of cancer cells define their sensitivity to HAMLET-induced death; c-Myc and glycolysis. AB - HAMLET is the first member of a new family of tumoricidal protein-lipid complexes that kill cancer cells broadly, while sparing healthy, differentiated cells. Many and diverse tumor cell types are sensitive to the lethal effect, suggesting that HAMLET identifies and activates conserved death pathways in cancer cells. Here, we investigated the molecular basis for the difference in sensitivity between cancer cells and healthy cells. Using a combination of small-hairpin RNA (shRNA) inhibition, proteomic and metabolomic technology, we identified the c-Myc oncogene as one essential determinant of HAMLET sensitivity. Increased c-Myc expression levels promoted sensitivity to HAMLET and shRNA knockdown of c-Myc suppressed the lethal response, suggesting that oncogenic transformation with c Myc creates a HAMLET-sensitive phenotype. Furthermore, HAMLET sensitivity was modified by the glycolytic state of tumor cells. Glucose deprivation sensitized tumor cells to HAMLET-induced cell death and in the shRNA screen, hexokinase 1 (HK1), 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 1 and hypoxia inducible factor 1alpha modified HAMLET sensitivity. HK1 was shown to bind HAMLET in a protein array containing ~8000 targets, and HK activity decreased within 15 min of HAMLET treatment, before morphological signs of tumor cell death. In parallel, HAMLET triggered rapid metabolic paralysis in carcinoma cells. Tumor cells were also shown to contain large amounts of oleic acid and its derivatives already after 15 min. The results identify HAMLET as a novel anti-cancer agent that kills tumor cells by exploiting unifying features of cancer cells such as oncogene addiction or the Warburg effect. PMID- 21643008 TI - Pdcd4 directly binds the coding region of c-myb mRNA and suppresses its translation. AB - Pdcd4 is a novel tumor suppressor protein that functions in the nucleus and the cytoplasm, and appears to be involved in the regulation of transcription and translation. In the cytoplasm, Pdcd4 has been implicated in the suppression of translation of mRNAs containing structured 5'-untranslated regions; however, the mechanisms that recruit Pdcd4 to specific target mRNAs and the identities of these mRNAs are mostly unknown. In this study, we have identified c-myb mRNA as the first natural translational target mRNA of Pdcd4. We have found that translational suppression of c-myb mRNA by Pdcd4 is dependent on sequences located within the c-myb-coding region. Furthermore, we have found that the N terminal domain of Pdcd4 has an important role in targeting Pdcd4 to c-myb RNA by mediating preferential RNA binding to the Pdcd4-responsive region of c-myb mRNA. Overall, our work demonstrates for the first time that Pdcd4 is directly involved in translational suppression of a natural mRNA and provides the first evidence for a key role of the RNA-binding domain in targeting Pdcd4 to a specific mRNA. PMID- 21643009 TI - IL-3 is a novel target to interfere with tumor vasculature. AB - Angiogenesis inhibiting agents are currently integral component of anticancer therapy. However, tumors, initially responsive to anti-angiogenic drugs or vascular targeting agents, can acquire resistance. The limited clinical efficacy might result from the heterogeneous nature of tumors or alternatively from the unique phenotype of tumor vascular cells, widely diverse from so-called 'normal' endothelium. Hence, defining the molecular mechanisms driving this diversity might provide a rational basis to design combinatory therapies that should be more effective in avoiding resistance. Herein, we demonstrated that tumor-derived endothelial cells (TECs) isolated from breast and kidney carcinomas retained an endothelial phenotype, but outspread independently of growth factors. Applying small interfering RNA approach, we demonstrated that interleukin (IL)-3, but not vascular endothelial growth factor, released by TECs, supports their autocrine growth and promotes in vivo vessel formation and tumor angiogenesis. Meanwhile, we found that the expression of the membrane-bound kit ligand (mbKitL) depends on IL-3, and it is crucial for adhesion of endothelial progenitor cells (EPCs) and inflammatory cells to TECs. These events required Akt activation. Finally, the finding that depletion of the mbKitL prevented EPC and inflammatory cell trafficking into vascular microenvironment, indicates that, as in bone marrow, the mbKitL can act as a membrane/adhesion molecule for c-Kit-expressing cells. These data provide evidences that an IL-3 autocrine loop can drive a tumor endothelial switch and that targeting IL-3 might confer a significant therapeutic advantage to hamper tumor angiogenesis. PMID- 21643010 TI - Inactivation of promoter 1B of APC causes partial gene silencing: evidence for a significant role of the promoter in regulation and causative of familial adenomatous polyposis. AB - Familial adenomatous polyposis (FAP) is caused by germline mutations in the adenomatous polyposis coli (APC) gene. Two promoters, 1A and 1B, have been recognized in APC, and 1B is thought to have a minor role in the regulation of the gene. We have identified a novel deletion encompassing half of this promoter in the largest family (Family 1) of the Swedish Polyposis Registry. The mutation leads to an imbalance in allele-specific expression of APC, and transcription from promoter 1B was highly impaired in both normal colorectal mucosa and blood from mutation carriers. To establish the significance of promoter 1B in normal colorectal mucosa (from controls), expression levels of specific transcripts from each of the promoters, 1A and 1B, were examined, and the expression from 1B was significantly higher compared with 1A. Significant amounts of transcripts generated from promoter 1B were also determined in a panel of 20 various normal tissues examined. In FAP-related tumors, the APC germline mutation is proposed to dictate the second hit. Mutations leaving two or three out of seven 20-amino-acid repeats in the central domain of APC intact seem to be required for tumorigenesis. We examined adenomas from mutation carriers in Family 1 for second hits in the entire gene without any findings, however, loss of the residual expression of the deleterious allele was observed. Three major conclusions of significant importance in relation to the function of APC can be drawn from this study; (i) germline inactivation of promoter 1B is disease causing in FAP; (ii) expression of transcripts from promoter 1B is generated at considerable higher levels compared with 1A, demonstrating a hitherto unknown importance of 1B; (iii) adenoma formation in FAP, caused by impaired function of promoter 1B, does not require homozygous inactivation of APC allowing for alternative genetic models as basis for adenoma formation. PMID- 21643011 TI - Phosphorylation and interaction of myopodin by integrin-link kinase lead to suppression of cell growth and motility in prostate cancer cells. AB - Myopodin is a tumor-suppressor gene that suppresses growth of prostate and urothelial carcinomas. However, the mechanism of myopodin tumor-suppressor activity or signaling that leads to activation of myopodin remains unclear. In this report, we showed that the N-terminus of myopodin binds integrin-linked kinase (ILK) both in vivo and in vitro. An ILK interaction motif of 78 amino acids (amino acids 82-157) was identified in the N-terminus region of myopodin. Induction of ILK-dependent kinase activity by integrin alpha7 led to phosphorylation of myopodin both in vivo and in vitro. Knocking down ILK dramatically reduced the inhibition of cell growth and motility mediated by myopodin. A mutant of myopodin lacking the ILK interaction motif is inactive in suppressing the growth and motility of PC3 cells. As a result, this study showed a novel and critical signaling pathway that leads to activation of myopodin. PMID- 21643012 TI - A novel oncogenic mechanism in Ewing sarcoma involving IGF pathway targeting by EWS/Fli1-regulated microRNAs. AB - MicroRNAs (miRs) are a novel class of cellular bioactive molecules with critical functions in the regulation of gene expression in normal biology and disease. MiRs are frequently misexpressed in cancer, with potent biological consequences. However, relatively little is known about miRs in pediatric cancers, including sarcomas. Moreover, the mechanisms behind aberrant miR expression in cancer are poorly understood. Ewing sarcoma is an aggressive pediatric malignancy driven by EWS/Ets fusion oncoproteins, which are gain-of-function transcriptional regulators. We employed stable silencing of EWS/Fli1, the most common of the oncogenic fusions, and global miR profiling to identify EWS/Fli1-regulated miRs with oncogenesis-modifying roles in Ewing sarcoma. In this report, we characterize a group of miRs (100, 125b, 22, 221/222, 27a and 29a) strongly repressed by EWS/Fli1. Strikingly, all of these miRs have predicted targets in the insulin-like growth factor (IGF) signaling pathway, a pivotal driver of Ewing sarcoma oncogenesis. We demonstrate that miRs in this group negatively regulate the expression of multiple pro-oncogenic components of the IGF pathway, namely IGF-1, IGF-1 receptor, mammalian/mechanistic target of rapamycin and ribosomal protein S6 kinase A1. Consistent with tumor-suppressive functions, these miRs manifest growth inhibitory properties in Ewing sarcoma cells. Our studies thus uncover a novel oncogenic mechanism in Ewing sarcoma, involving post transcriptional derepression of IGF signaling by the EWS/Fli1 fusion oncoprotein via miRs. This novel pathway may be amenable to innovative therapeutic targeting in Ewing sarcoma and other malignancies with activated IGF signaling. PMID- 21643013 TI - Cancer cells in epithelial-to-mesenchymal transition and tumor-propagating-cancer stem cells: distinct, overlapping or same populations. AB - Cell populations of solid cancers and their distant models, the cancer cell lines, have been categorized in sub-populations: cancer stem-tumor-propagating cells (CSC-TPC) versus derived cells, epithelial- versus mesenchymal-type cells, dormant versus actively proliferating cells and so on. CSC-TPC are minimally defined by their operational properties: immortality and the ability to regenerate in vivo or in vitro the whole panel of cancer cells. The epithelial-to mesenchymal transition (EMT), mostly observed in vitro, generates mesenchymal type from epithelial-type cells. The converse transition is mesenchymal-to epithelial transition. In vitro work suggests that CSC-TPC and EMT cell phenotypes overlap. An analysis of the properties of these sub-populations, as studied in vitro, shows that indeed these two phenotypes may be linked to some extent. However, the in vivo counterpart of this relation in human tumors has barely been investigated. A model in which among the EMT cells released from the tumor only the most competent CSC-TPC will succeed to metastasize is proposed. It is suggested that in the Darwinian evolution of cancer cells, many phenotypes reflecting the expression of various programs, reversible to irreversible, exclusive, overlapping or linked coexist and compete with each other. PMID- 21643014 TI - The tumor-suppressor gene ARHI (DIRAS3) suppresses ovarian cancer cell migration through inhibition of the Stat3 and FAK/Rho signaling pathways. AB - Ovarian cancers migrate and metastasize over the surface of the peritoneal cavity. Consequently, dysregulation of mechanisms that limit cell migration may be particularly important in the pathogenesis of the disease. ARHI is an imprinted tumor-suppressor gene that is downregulated in >60% of ovarian cancers, and its loss is associated with decreased progression-free survival. ARHI encodes a 26-kDa GTPase with homology to Ras. In contrast to Ras, ARHI inhibits cell growth, but whether it also regulates cell motility has not been studied previously. Here we report that re-expression of ARHI decreases the motility of IL-6- and epidermal growth factor (EGF)-stimulated SKOv3 and Hey ovarian cancer cells, inhibiting both chemotaxis and haptotaxis. ARHI binds to and sequesters Stat3 in the cytoplasm, preventing its translocation to the nucleus and localization in focal adhesion complexes. Stat3 siRNA or the JAK2 inhibitor AG490 produced similar inhibition of motility. However, the combination of ARHI expression with Stat3 knockdown or inhibition produced greatest inhibition in ovarian cancer cell migration, consistent with Stat3-dependent and Stat3 independent mechanisms. Consistent with two distinct signaling pathways, knockdown of Stat3 selectively inhibited IL-6-stimulated migration, whereas knockdown of focal adhesion kinase (FAK) preferentially inhibited EGF-stimulated migration. In EGF-stimulated ovarian cancer cells, re-expression of ARHI inhibited FAK(Y397) and Src(Y416) phosphorylation, disrupted focal adhesions, and blocked FAK-mediated RhoA signaling, resulting in decreased levels of GTP-RhoA. Re-expression of ARHI also disrupted the formation of actin stress fibers in a FAK- and RhoA-dependent manner. Thus, ARHI has a critical and previously uncharacterized role in the regulation of ovarian cancer cell migration, exerting inhibitory effects on two distinct signaling pathways. PMID- 21643015 TI - Regulation of mitosis and taxane response by Daxx and Rassf1. AB - Current theories suggest that mitotic checkpoint proteins are essential for proper cellular response to taxanes, a widely used family of chemotherapeutic compounds. We recently showed that absence or depletion of protein Daxx increases cellular taxol (paclitaxel) resistance-a common trait of patients diagnosed with several malignancies, including breast cancer. Further investigation of Daxx mediated taxol response revealed that Daxx is important for the proper timing of mitosis progression and cyclin B stability. Daxx interacts with mitotic checkpoint protein RAS-association domain family protein 1 (Rassf1) and partially colocalizes with this protein during mitosis. Rassf1/Daxx depletion or expression of Daxx-binding domain of Rassf1 elevates cyclin B stability and increases taxol resistance in cells and mouse xenograft models. In breast cancer patients, we observed the inverse correlation between Daxx and clinical response to taxane based chemotherapy. These data suggest that Daxx and Rassf1 define a mitotic stress checkpoint that enables cells to exit mitosis as micronucleated cells (and eventually die) when encountered with specific mitotic stress stimuli, including taxol. Surprisingly, depletion of Daxx or Rassf1 does not change the activity of E3 ubiquitin ligase anaphase promotion complex/C in in vitro settings, suggesting the necessity of mitotic cellular environment for proper activation of this checkpoint. Daxx and Rassf1 may become useful predictive markers for the proper selection of patients for taxane chemotherapy. PMID- 21643016 TI - miR-296 regulation of a cell polarity-cell plasticity module controls tumor progression. AB - The expression of small, non-coding RNA or microRNAs (miR), is frequently deregulated in human cancer, but how these pathways affect disease progression is still largely elusive. Here, we report on a miR, miR-296, which is progressively lost during tumor progression and correlates with metastatic disease in colorectal, breast, lung, gastric, parathyroid, liver and bile ducts cancers. Functionally, miR-296 controls a global cell motility gene signature in epithelial cells by transcriptionally repressing the cell polarity-cell plasticity module, Scribble (Scrib). In turn, loss of miR-296 causes aberrantly increased and mislocalized Scrib in human tumors, resulting in exaggerated random cell migration and tumor cell invasiveness. Re-expression of miR-296 in MDA-MB231 cells inhibits tumor growth in vivo. Finally, miR-296 or Scrib levels predict tumor relapse in hepatocellular carcinoma patients. These data identify miR-296 as a global repressor of tumorigenicity and uncover a previously unexplored exploitation of Scrib in tumor progression in humans. PMID- 21643017 TI - MiR-100 regulates cell differentiation and survival by targeting RBSP3, a phosphatase-like tumor suppressor in acute myeloid leukemia. AB - Acute myeloblastic leukemia (AML) is characterized by the accumulation of abnormal myeloblasts (mainly granulocyte or monocyte precursors) in the bone marrow and blood. Though great progress has been made for improvement in clinical treatment during the past decades, only minority with AML achieve long-term survival. Therefore, further understanding mechanisms of leukemogenesis and exploring novel therapeutic strategies are still crucial for improving disease outcome. MicroRNA-100 (miR-100), a small non-coding RNA molecule, has been reported as a frequent event aberrantly expressed in patients with AML; however, the molecular basis for this phenotype and the statuses of its downstream targets have not yet been elucidated. In the present study, we found that the expression level of miR-100 in vivo was related to the stage of the maturation block underlying the subtypes of myeloid leukemia. In vitro experiments further demonstrated that miR-100 was required to promote the cell proliferation of promyelocytic blasts and arrest them differentiated to granulocyte/monocyte lineages. Significantly, we identified RBSP3, a phosphatase-like tumor suppressor, as a bona fide target of miR-100 and validated that RBSP3 was involved in cell differentiation and survival in AML. Moreover, we revealed a new pathway that miR-100 regulates G1/S transition and S-phase entry and blocks the terminal differentiation by targeting RBSP3, which partly in turn modulates the cell cycle effectors pRB/E2F1 in AML. These events promoted cell proliferation and blocked granulocyte/monocyte differentiation. Our data highlight an important role of miR-100 in the molecular etiology of AML, and implicate the potential application of miR-100 in cancer therapy. PMID- 21643018 TI - Acquisition of p53 mutations in response to the non-genotoxic p53 activator Nutlin-3. AB - Wild-type p53 is a stress-responsive tumor suppressor and potent growth inhibitor. Genotoxic stresses (for example, ionizing and ultraviolet radiation or chemotherapeutic drug treatment) can activate p53, but also induce mutations in the P53 gene, and thus select for p53-mutated cells. Nutlin-3a (Nutlin) is pre clinical drug that activates p53 in a non-genotoxic manner. Nutlin occupies the p53-binding pocket of murine double minute 2 (MDM2), activating p53 by blocking the p53-MDM2 interaction. Because Nutlin neither binds p53 directly nor introduces DNA damage, we hypothesized Nutlin would not induce P53 mutations, and, therefore, not select for p53-mutated cells. To test this, populations of SJSA-1 (p53 wild-type) cancer cells were expanded that survived repeated Nutlin exposures, and individual clones were isolated. Group 1 clones were resistant to Nutlin-induced apoptosis, but still underwent growth arrest. Surprisingly, while some Group 1 clones retained wild-type p53, others acquired a heterozygous p53 mutation. Apoptosis resistance in Group 1 clones was associated with decreased PUMA induction and decreased caspase 3/7 activation. Group 2 clones were resistant to both apoptosis and growth arrest induced by Nutlin. Group 2 clones had acquired mutations in the p53-DNA-binding domain and expressed only mutant p53s that were induced by Nutlin treatment, but were unable to bind the P21 and PUMA gene promoters, and unable to activate transcription. These results demonstrate that non-genotoxic p53 activation (for example, by Nutlin treatment) can lead to the acquisition of somatic mutations in p53 and select for p53 mutated cells. These findings have implications for the potential clinical use of Nutlin and other small molecule MDM2 antagonists. PMID- 21643019 TI - TP63 P2 promoter functional analysis identifies beta-catenin as a key regulator of DeltaNp63 expression. AB - The DeltaNp63 protein, a product of the TP63 gene that lacks the N-terminal domain, has a critical role in the maintenance of self renewal and progenitor capacity in several types of epithelial tissues. DeltaNp63 is frequently overexpressed in squamous cell carcinoma (SCC) and in some other epithelial tumours. This overexpression may contribute to tumour progression through dominant-negative effects on the transcriptionally active (TA) isoforms of the p53 family (TAp63, TAp73 and p53), as well as through independent mechanisms. However, the molecular basis of DeltaNp63 overexpression is not fully understood. Here, we show that the expression of DeltaNp63 is regulated by the Wnt/beta catenin pathway in human hepatocellular carcinoma (HCC) and SCC cell lines. This regulation operates in particular through TCF/LEF sites present in the P2 promoter of TP63. In addition, we show that DeltaNp63 and beta-catenin are frequently coexpressed and accumulated in oesophageal SCC, but not in HCC. These results suggest that activation of the beta-catenin pathway may contribute to overexpression of DeltaNp63 during tumour progression, in a cell type-specific manner. PMID- 21643020 TI - A prognostic score with pretransplant serum ferritin and disease status predicts outcome following reduced-intensity SCT. PMID- 21643021 TI - The EBMT risk score. AB - The European Group for Blood and Marrow Transplantation (EBMT) risk score provides a simple tool to assess instantly chances and risks of hematopoietic SCT(HSCT) for an individual patient pre-transplant. Five factors, age of the patient, stage of the disease, time from diagnosis, donor type and donor recipient gender combination augment risk for an individual patient with increasing score from 0 as best to 7 as worst in an additive way. The score holds for all acquired hematological disorders, for allogeneic and autologous HSCT (score 0-5), is independent of the HSCT technology and is valid for standard or reduced intensity conditioning. Survival is uniformly worse for older patients, transplanted in advanced disease stage after a long-time interval and with a mismatched donor than for younger patients, transplanted soon in early stage with a well matched donor. Additional risk factors such as performance score, CMV serostatus or cytokine polymorphisms improve prediction but to different extents for low or high-risk patients. Comparative assessment of disease risk and global pre-transplant risk should guide decisions for each patient with his/her specific disease between HSCT and a non-transplant approach and replace the traditional 'donor vs no donor' with such a risk-adapted individualized strategy. PMID- 21643022 TI - Metabolic syndrome and cardiovascular risk in survivors after hematopoietic cell transplantation. AB - Increasing numbers of hematopoietic cell transplantations (HCTs) are being performed annually with a greater number of long-term survivors. There is increasing concern regarding the late complications and long-term effects that are secondary to treatment exposures before HCT as well as during the HCT conditioning therapy. In both the autologous as well as allogeneic transplant setting, transplant survivors experience mortality rates higher than the general population and the risk of premature cardiovascular (CV)-related death is increased 2.3-fold compared with the general population. The etiology of CV related deaths in HCT survivors is multifactorial; however, increasing evidence suggests that HCT survivors are at higher risk of developing adverse CV risk factors leading to the development of the metabolic syndrome (a constellation high triglyceride levels, low high-density lipoprotein-cholesterol, hypertension, high fasting blood sugars and increased waist circumference), which then predisposes individuals to risk for early CV-related death. Resistance to insulin is the primary underlying pathophysiologic mechanism that contributes to the development of metabolic syndrome and HCT survivors have been shown to be more likely to develop hypertension, hyperlipidemia and to be insulin resistant. However, the relationship between HCT-related treatment exposures (total body irradiation, high dose chemotherapy, calcineurin inhibitors, steroids, etc) and transplant-related complications (such as GVHD) with the development of CV risk factors and insulin resistance is still in the early stages of investigation. Greater knowledge of the concern regarding CV risk in HCT survivors among both patients and care providers will provide the opportunity for appropriate screening as well as interventions for modifiable risk factors. PMID- 21643023 TI - Wilms' tumor gene 1 expression: an independent acute leukemia prognostic indicator following allogeneic hematopoietic SCT. AB - To evaluate the prognostic significance of Wilms' tumor gene 1 (WT1) expression for monitoring minimal residual disease and predicting relapse in patients with acute leukemia (AL) following allogeneic hematopoietic SCT (allo-HSCT), the WT1 expression levels of 138 AL patients were measured using real-time quantitative reverse transcription PCR at designed time points after allo-HSCT. All patients were divided into four groups based on the HSCT outcomes and intervention application. A low level of WT1 expression following HSCT indicated a low risk of relapse, whereas WT1 expression >1.05% was indicative of a higher probability of relapse. Only the advanced stage of disease (hazard ratio (HR)=2.73; 95% confidence interval (CI)=1.337-5.573, P=0.006) and a WT1 expression >= 0.60% (HR=4.774; 95% CI=2.410-9.459, P=0.000) were associated with lower disease-free survival. Relapse (HR=0.119; 95% CI=0.056-0.250, P=0.000) and a WT1 expression ?0.60% (HR=2.771; 95% CI=1.316-5.834, P=0.007) were associated with lower OS. In conclusion, the WT1 expression level is an independent prognostic factor that can predict clinical outcomes for AL patients after HSCT and provide a guide for suitable interventions. PMID- 21643024 TI - Obesity: Potential value of combined treatment for the medical management of obesity. PMID- 21643026 TI - Hepatitis C: Induction of immune responses against HCV. PMID- 21643027 TI - Infection: Helicobacter pylori may protect against IBD--a mechanistic insight. PMID- 21643028 TI - Cancer: CD40 agonists--a promising new treatment for pancreatic cancer? PMID- 21643029 TI - Gastrointestinal bleeding: Novel nanopowder spray shows promising results in humans for the treatment of acute peptic ulcer bleeding. PMID- 21643030 TI - Cancer: Gene fusion identified in gastric cancer. PMID- 21643032 TI - PPI therapy: Clopidrogel-PPI drug interaction may not be a class effect. PMID- 21643033 TI - Pain: Transcranial direct current stimulation reduces post-ERCP pain. PMID- 21643034 TI - Ulcerative colitis: Rescue immune suppression for severe colitis--worth the risk? PMID- 21643035 TI - Hepatitis: PEG-IFN for the treatment of hepatitis D. PMID- 21643036 TI - Constipation: Dried plums (prunes) for the treatment of constipation. PMID- 21643037 TI - Helicobacter pylori eradication: A new, single-capsule bismuth-containing quadruple therapy. PMID- 21643038 TI - Influence of obesity on the risk of esophageal disorders. AB - Obesity is associated with an increased risk of esophageal disorders, including esophageal adenocarcinoma, Barrett esophagus and GERD. For reasons yet unknown, the association between obesity and esophageal adenocarcinoma seems to be stronger than that for other types of obesity-related cancers. Predominantly abdominal or intra-abdominal adiposity (representing visceral fat and other fat within the abdominal cavity), which is more frequently observed in men than in women, is more strongly linked with these esophageal disorders than BMI alone, a finding that might contribute to the striking male predominance of esophageal adenocarcinoma. Research has identified potential mechanisms underlying the strong link between obesity and esophageal conditions. These findings are summarized in this Review, but more research remains to be carried out before these mechanisms are established. PMID- 21643039 TI - New insights into visceral hypersensitivity--clinical implications in IBS. AB - A subset of patients with IBS have visceral hypersensitivity and/or somatic hypersensitivity. Visceral hypersensitivity might have use as a clinical marker of IBS and could account for symptoms of urgency for bowel movements, bloating and abdominal pain. The mechanisms that lead to chronic visceral hypersensitivity in patients who have IBS are unclear. However, several working models may be considered, including: nociceptive input from the colon that leads to hypersensitivity; increased intestinal permeability that induces a visceral nociceptive drive; and alterations in the expression of microRNAs in gastrointestinal tissue that might be delivered via blood microvesicles to other target organs, such as the peripheral and/or central nervous system. As such, the chronic visceral hypersensitivity that is present in a subset of patients with IBS might be maintained by both peripheral and central phenomena. The theories underlying the development of chronic visceral hypersensitivity in patients with IBS are supported by findings from new animal models in which hypersensitivity follows transient inflammation of the colon. The presence of somatic hypersensitivity and an alteration in the neuroendocrine system in some patients who have IBS suggests that multisystemic factors are involved in the overall disorder. Thus, IBS is similar to other chronic pain disorders, such as fibromyalgia, chronic regional pain disorder and temporomandibular joint disorder, as chronic nociceptive mechanisms are activated in all of these disorders. PMID- 21643040 TI - Plant virology: Sign turns yellow for Y-sat-infected tobacco. PMID- 21643042 TI - Direct femtosecond laser waveguide writing inside zinc phosphate glass. AB - We report the relationship between the initial glass composition and the resulting microstructural changes after direct femtosecond laser waveguide writing with a 1 kHz repetition rate Ti:sapphire laser system. A zinc polyphosphate glass composition with an oxygen to phosphorus ratio of 3.25 has demonstrated positive refractive index changes induced inside the focal volume of a focusing microscope objective for laser pulse energies that can achieve intensities above the modification threshold. The permanent photo-induced changes can be used for direct fabrication of optical waveguides using single scan writing techniques. Changes to the localized glass network structure that produce positive changes in the refractive index of zinc phosphate glasses upon femtosecond laser irradiation have been studied using scanning confocal micro Raman and fluorescence spectroscopy. PMID- 21643041 TI - How does a hypha grow? The biophysics of pressurized growth in fungi. AB - The mechanisms underlying the growth of fungal hyphae are rooted in the physical property of cell pressure. Internal hydrostatic pressure (turgor) is one of the major forces driving the localized expansion at the hyphal tip which causes the characteristic filamentous shape of the hypha. Calcium gradients regulate tip growth, and secretory vesicles that contribute to this process are actively transported to the growing tip by molecular motors that move along cytoskeletal structures. Turgor is controlled by an osmotic mitogen-activated protein kinase cascade that causes de novo synthesis of osmolytes and uptake of ions from the external medium. However, as discussed in this Review, turgor and pressure have additional roles in hyphal growth, such as causing the mass flow of cytoplasm from the basal mycelial network towards the expanding hyphal tips at the colony edge. PMID- 21643043 TI - High sensitivity SMS fiber structure based refractometer--analysis and experiment. AB - We have investigated the influence of multimode fiber core (MMFC) diameters and lengths on the sensitivity of an SMS fiber based refractometer. We show that the MMFC diameter has significant influence on the refractive index (RI) sensitivity but the length does not. A refractometer with a lower MMFC diameter has a higher sensitivity. Experimental investigations achieved a maximum sensitivity of 1815 nm/ RIU (refractive index unit) for a refractive index range from 1.342 to 1.437 for a refractometer with a core diameter of 80 MUm. The experimental results fit well with the numerical simulation results. PMID- 21643044 TI - Effects of particle aggregation and disaggregation on their inherent optical properties. AB - In many environments a large portion of particulate material is contained in aggregated particles; however, there is no validated framework to describe how aggregates in the ocean scatter light. Here we present the results of two experiments aiming to expose the role that aggregation plays in determining particle light scattering properties, especially in sediment-dominated coastal waters. First, in situ measurements of particle size distribution (PSD) and beam attenuation were made with two laser particle sizing instruments (one equipped with a pump to subject the sample to aggregate-breaking shear), and measurements from the two treatments were compared. Second, clays were aggregated in the laboratory using salt, and observed over time by multiple instruments in order to examine the effects of aggregation and settling on spectral beam-attenuation and backscattering. Results indicate: (1) mass normalized attenuation and backscattering are only weakly sensitive to size changes due to aggregation in contrast to theory based on solid particles, (2) the spectral slope of beam attenuation is indicative of changes in PSD but is complicated by instrument acceptance angle, and (3) the spectral shape of backscattering did not provide as clear a relationship with PSD as spectral beam attenuation, as is predicted by theory for solid spheres. PMID- 21643045 TI - Coherent anti-Stokes Raman scattering microscopy imaging with suppression of four wave mixing in optical fibers. AB - We demonstrated an optical fiber delivered coherent anti-Stokes Raman scattering (CARS) microscopy imaging system with a polarization-based mechanism for suppression of four-wave mixing (FWM) signals in delivery fiber. Polarization maintaining fibers (PMF) were used as the delivery fiber to ensure stability of the state of polarization (SOP) of lasers. The pump and Stokes waves were coupled into PMFs at orthogonal SOPs along the slow and fast axes of PMFs, respectively, resulting in a significant reduction of FWM signals generated in the fiber. At the output end of PMFs, a dual-wavelength waveplate was used to realign the SOPs of the two waves into identical SOPs prior to their entrance into the CARS microscope. Therefore, it allows the pump and Stokes waves with identical SOPs to excite samples at highest excitation efficiency. Our experimental results showed that this polarization-based FWM-suppressing mechanism can dramatically reduce FWM signals generated in PMFs up to approximately 99%. Meanwhile, the PMF delivered CARS microscopy system with this mechanism can still produce high quality CARS images. Consequently, our PMF-delivered CARS microscopy imaging system with the polarization-based FWM-suppressing mechanism potentially offers a new strategy for building fiber-based CARS endoscopes with effective suppression of FWM background noises. PMID- 21643046 TI - Detection of bacteria using bacteriophages as recognition elements immobilized on long-period fiber gratings. AB - The paper presents for the first time a study of long-period gratings (LPGs) applied for label-free detection of specific bacteria using physically adsorbed bacteriophages. For the purposes of the experiment a number of highly sensitive LPGs working at the turning point of phase matching curve was fabricated in SMF28 fiber using UV exposure. We show that the device allows for real-time monitoring of phenomena taking place on the sensor's surface, including phage-bacteria interactions. For the applied conditions a resonance wavelength shift of ~1.3 nm induced by bacteria binding was observed. PMID- 21643047 TI - Electro-optic effect in crystalline films of transverse planar octupolar symmetry. AB - We present theoretical and experimental demonstrations of the electro-optic activity in crystalline molecular thin films with octupolar D(3h) symmetry. Applying a longitudinal electric field modulation within the molecular plane, we analyze the induced refractive index change relative to the orientation of the octupoles in their plane, and show that a maximum value is reached when one octupolar branch lies along the direction of the modulating field. These characteristics, as well as their electric field dependence, are drastically different from more traditional one-dimensional symmetry samples, bringing additional advantages related to electro-optic coupling possibilities. PMID- 21643048 TI - Tm3+-doped CW fiber laser based on a highly GeO2-doped dispersion-shifted fiber. AB - A novel all-fiber laser based on a highly GeO2-doped dispersion-shifted Tm codoped fiber, pumped at 1.56 um wavelength and lasing at 1.862 um wavelength with a slope efficiency up to 37% was demonstrated. The single-mode Tm-doped fiber with the 55GeO2-45SiO2 core was fabricated for the first time by MCVD technique. The laser produces spectral side bands, resulting from the four-wave mixing owing to the shift of the zero-dispersion-wavelength of the fiber to the laser wavelength, thus, making it potentially particularly attractive for dispersion management and ultrashort pulse generation. PMID- 21643049 TI - An upstream multi-wavelength shared PON based on tunable self-seeding Fabry-Perot laser diode for upstream capacity upgrade and wavelength multiplexing. AB - We proposed an Upstream Multi-Wavelength Shared (UMWS) PON architecture based on a tunable self-seeding Fabry-Perot laser diode (FP-LD) at ONU. The performances of the wavelength and power stability, side-mode suppression ratio (SMSR), tuning range for the proposed tunable self-seeding laser module at ONU are experimentally investigated. The BER is measured with direct modulation on FP-LD of 1.25 Gbps upstream data. The extensive simulations not only evaluate the enhanced performance from the upstream wavelength-sharing, but also for the first time investigate the impact of channel Switch Latency (SL) on the network performance. PMID- 21643050 TI - Imaging properties of supercritical angle fluorescence optics. AB - In recent years, new optical systems have been developed with the ability to collect light at very high angles of emission, exceeding the critical angle of total internal reflection. Prominent examples are solid-immersion lenses and paraboloid collectors. These systems achieve high efficiencies in fluorescence detection which is an important issue for sensitive applications in analytical chemistry and biochemical assays. The exclusive collection of supercritical angle fluorescence (SAF) allows for the detection of evanescent modes and thus to confine the detection volume within one wavelength to an interface. For conventional optical systems with high numerical aperture a precise wave-optical theory of imaging was developed by Richards and Wolf in the fifties of the last century. However, their theory is not directly applicable to non-imaging, strongly aberratic light collection systems systems that collect a significant part of light above the critical angle. Here, we extend the theory to describe the optical properties of such systems. PMID- 21643051 TI - Compression of digital hologram for three-dimensional object using Wavelet Bandelets transform. AB - In the transformation based compression algorithms of digital hologram for three dimensional object, the balance between compression ratio and normalized root mean square (NRMS) error is always the core of algorithm development. The Wavelet transform method is efficient to achieve high compression ratio but NRMS error is also high. In order to solve this issue, we propose a hologram compression method using Wavelet-Bandelets transform. Our simulation and experimental results show that the Wavelet-Bandelets method has a higher compression ratio than Wavelet methods and all the other methods investigated in this paper, while it still maintains low NRMS error. PMID- 21643052 TI - Waveguide self-coupling based reconfigurable resonance structure for optical filtering and delay. AB - We propose a novel waveguide self-coupling based reconfigurable resonance structure that works as a flat-top second-order tunable filter and a tunable delay line with low group delay dispersion. The high-order resonance features result from the mutual mode coupling between the clockwise and counter-clockwise resonance eigenmodes. The transfer-matrix method is used to theoretically analyze the device optical performances. The relations between the two embedded phase shifters for achieving flat-top filtering and group delay responses are given. As the coupled resonances are provided by only one physical resonator, the device is inherently more compact and resilient to fabrication errors compared to conventional microring resonators. Phase tuning for its reconfiguration is also simpler and more power-efficient. PMID- 21643053 TI - Polarization independent adaptive microlens with a blue-phase liquid crystal. AB - A new polarization-independent and fast-response adaptive microlens array using a polymer-stabilized blue phase liquid crystal is proposed. With a curved top electrode and planar bottom electrode, gradient electric fields are generated and lens-like phase profile obtained. Optimization process leads to an ideal parabolic phase profile for suppressing spherical aberration. PMID- 21643054 TI - Measuring translational, rotational, and vibrational dynamics in colloids with digital holographic microscopy. AB - We discuss a new method for simultaneously probing translational, rotational, and vibrational dynamics in dilute colloidal suspensions using digital holographic microscopy (DHM). We record digital holograms of clusters of 1-MUm-diameter colloidal spheres interacting through short-range attractions, and we fit the holograms to an exact model of the scattering from multiple spheres. The model, based on the T-matrix formulation, accounts for multiple scattering and near field coupling. We also explicitly account for the non-asymptotic radial decay of the scattered fields, allowing us to accurately fit holograms recorded with the focal plane located as little as 15 MUm from the particle. Applying the fitting technique to a time-series of holograms of Brownian dimers allows simultaneous measurement of six dynamical modes - three translational, two rotational, and one vibrational - on timescales ranging from 10(-3) to 1 s. We measure the translational and rotational diffusion constants to a precision of 0.6%, and we use the vibrational data to measure the interaction potential between the spheres to a precision of ~50 nm in separation distance. Finally, we show that the fitting technique can be used to measure dynamics of clusters containing three or more spheres. PMID- 21643055 TI - STED nanoscopy with mass-produced laser diodes. AB - We show that far-field fluorescence nanoscopy by stimulated emission depletion (STED) can be realized with compact off-the-shelf laser diodes, such as those used in laser pointers and DVDs. A spatial resolution of 40-50 nm is attained by pulsing a 660 nm DVD-diode. The efficacy of these low-cost STED microscopes in biological imaging is demonstrated by differentiating between clusters of the synaptic protein bassoon and transport vesicles in hippocampal neurons, based on the feature diameter. Our results facilitate the implementation of this all molecular-transition based superresolution method in many applications ranging from nanoscale fluorescence imaging to nanoscale fluorescence sensing. PMID- 21643056 TI - Experimental characterization of the coherence properties of hard x-ray sources. AB - The experimental characterization of the coherence properties of hard X-ray sources is reported and discussed. The source is described by its Mutual Optical Intensity (MOI). The coherent-mode decomposition is applied to the MOI described by a Gaussian-Schell model. The method allows for a direct, quantitative characterization of the degree of coherence of both synchrotron and laboratory sources. The latter represents the first example of characterizing a low coherence hard x-ray source. PMID- 21643057 TI - Optimizing the subcarrier granularity of coherent optical communications systems. AB - In this paper, we use numerical simulations to show that the symbol rate has a significant effect on the nonlinearity-limited performance of coherent optical communication systems. We consider the case where orthogonal subcarriers are used to maximize the spectral efficiency. Symbol rates from 0.78125 Gbaud to 100 Gbaud and links of up to 3200 km, without inline dispersion compensation, were simulated. The results show that the optimal symbol rates for the 800-km link and 3200-km link were 6.25-Gbaud and 3.125-Gbaud respectively. The optimal baud rate decreases as the length of the link is increased. After 3200 km, the performance of the 100-Gbaud system was worst in the nonlinearity-limited regime producing a received Q 2.4-dB lower than the 3.125-Gband system. The variation in the nonlinearity-limited performance is explained by using Cross-Phase-Modulation (XPM) theory and by considering the RF spectra of the intensity fluctuations of the signal along the link. The findings of the paper suggest that the maximum capacity of nonlinear dispersive optical links can only be achieved by using multiple subcarriers carrying a few Gbaud each, and not by high symbol rate systems. PMID- 21643058 TI - High performance transflective liquid crystal display associated with fringe field switching device. AB - The outdoor readability of the most popular portable liquid crystal display (LCDs) viz. fringe field switching has been addressed both in single and dual cell gap transflective devices. The devices use dual orientation, such as, homogeneous alignment in transmissive (T) part and 64 degrees twisted alignment in reflective (R) part. The dark states of the proposed devices are achieved by controlling phase retardation in T part and polarization rotation in R part and the white state is realized by rotating optic axis of liquid crystal and removing phase retardation in T and R parts, respectively. The devices show high light efficiency without requiring any optical compensation films, exhibiting strong potential for portable display applications. PMID- 21643059 TI - Frequency domain analysis for laser-locked cavity ringdown spectroscopy. AB - In this paper we report on the development of a Fourier-transform based signal processing method for laser-locked Continuous Wave Cavity Ringdown Spectroscopy (CWCRDS). Rather than analysing single ringdowns, as is the norm in traditional methods, we amplitude modulate the incident light, and analyse the entire waveform output of the optical cavity; our method has more in common with Cavity Attenuated Phase Shift Spectroscopy than with traditional data analysis methods. We have compared our method to Levenburg-Marquardt non linear least squares fitting, and have found that, for signals with a noise level typical of that from a locked CWCRDS instrument, our method has a comparable accuracy and comparable or higher precision. Moreover, the analysis time is approximately 500 times faster (normalised to the same number of time domain points). Our method allows us to analyse any number of periods of the ringdown waveform at once: this allows the method to be optimised for speed and precision for a given spectrometer. PMID- 21643060 TI - Analysis and engineering of chromatic dispersion in silicon waveguide bends and ring resonators. AB - We analyze chromatic dispersion in tightly curved silicon strip and slot waveguides with high index contrast. It is found that the dispersion profile is changed dramatically at both polarization states, when bending radius is reduced to a few microns. Zero-dispersion wavelength may shift by more than 220 nm, which raises a critical issue in design and optimization of micro-resonator-based devices for nonlinear applications. We propose a slot structure to tailor in cavity dispersion and obtain spectral lines with the standard deviation of frequency-dependent free spectral range of the slot-waveguide resonator made 460 times smaller than that of a strip-waveguide resonator, making it suitable for on chip octave-spanning frequency comb generation in mid-infrared wavelength range. PMID- 21643061 TI - Femtosecond high-power quantum dot vertical external cavity surface emitting laser. AB - We report on the first femtosecond vertical external cavity surface emitting laser (VECSEL) exceeding 1 W of average output power. The VECSEL is optically pumped, based on self-assembled InAs quantum dot (QD) gain layers, cooled efficiently using a thin disk geometry and passively modelocked with a fast quantum dot semiconductor saturable absorber mirror (SESAM). We developed a novel gain structure with a flat group delay dispersion (GDD) of +/- 10 fs2 over a range of 30 nm around the designed operation wavelength of 960 nm. This amount of GDD is several orders of magnitude lower compared to standard designs. Furthermore, we used an optimized positioning scheme of 63 QD gain layers to broaden and flatten the spectral gain. For stable and self-starting pulse formation, we have employed a QD-SESAM with a fast absorption recovery time of around 500 fs. We have achieved 1 W of average output power with 784-fs pulse duration at a repetition rate of 5.4 GHz. The QD-SESAM and the QD-VECSEL are operated with similar cavity mode areas, which is beneficial for higher repetition rates and the integration of both elements into a modelocked integrated external-cavity surface emitting laser (MIXSEL). PMID- 21643062 TI - Phase-sensitive swept-source interferometry for absolute ranging with application to measurements of group refractive index and thickness. AB - Interferometric range measurements using a wavelength-tunable source form the basis of several measurement techniques, including optical frequency domain reflectometry (OFDR), swept-source optical coherence tomography (SS-OCT), and frequency-modulated continuous wave (FMCW) lidar. We present a phase-sensitive and self-referenced approach to swept-source interferometry that yields absolute range measurements with axial precision three orders of magnitude better than the transform-limited axial resolution of the system. As an example application, we implement the proposed method for a simultaneous measurement of group refractive index and thickness of an optical glass sample. PMID- 21643063 TI - Quantitative phase imaging with polychromatic x-ray sources. AB - We introduce theoretically and demonstrate experimentally a contrast transfer function based phase retrieval algorithm that reconstructs the projected thickness of an homogeneous sample using a polychromatic x-ray source. We show excellent quantitative recovery of test samples in 2D using a synchrotron source with significant harmonic contamination, and in 3D using a laboratory source. PMID- 21643064 TI - Laboratory demonstrations on a pyramid wavefront sensor without modulation for closed-loop adaptive optics system. AB - The feasibility and performance of the pyramid wavefront sensor without modulation used in closed-loop adaptive optics system is investigated in this paper. The theory concepts and some simulation results are given to describe the detection trend and the linearity range of such a sensor with the aim to better understand its properties, and then a laboratory setup of the adaptive optics system based on this sensor and the liquid-crystal spatial light modulator is built. The correction results for the individual Zernike aberrations and the Kolmogorov phase screens are presented to demonstrate that the pyramid wavefront sensor without modulation can work as expected for closed-loop adaptive optics system. PMID- 21643065 TI - Correlation between color quality metric predictions and visual appreciation of light sources. AB - Over the past years there has been increasing evidence that the CIE color rendering index R(a) fails to correspond to the perceived color quality of many light sources, especially some Light-Emitting-Diodes. Several proposals to update, complement or even replace the CIE R(a) have therefore been made. The performance of thirteen color quality metrics was evaluated by calculating the average correlation of the metric predictions with the visual scaling of the perceived color quality obtained in several psychophysical studies. Two aspects of perceived color quality were investigated, appreciation (preference or attractiveness) and naturalness. The memory color quality metric (S(a)) of Smet et al. was found to correlate highly with perceived appreciation (r = 0.88). It was found to be statistically better (p<0.0001) at it than all other metrics. The CIE R(a) performed the worst. A metric that combines the gamut area index (GAI) and the CIE R(a) using an arithmetic mean correlated highly with the perceived naturalness of a light source (r = 0.85). It was found to be statistically better at predicting naturalness than all other metrics (p<0.0001). A negative correlation was found, between the capabilities of a light source's ability to predict appreciation and naturalness, indicating that a complete description of the color quality of a light source probably requires more than one metric. PMID- 21643066 TI - Mode-beating-enabled stopband narrowing in all-solid photonic bandgap fiber and sensing applications. AB - In this paper, core-cladding modal beating in a short piece of all-solid photonic bandgap fiber (AS-PBF) is observed in longitudinal propagation direction. It is demonstrated that at the stopband range of AS-PBF, the power could transfer back and forth between the fiber core and the first layer of high-index rods. Both experimental results and the theoretical analysis from transverse coupled mode theory confirm that the 3-dB width of the sharp stopband could be significantly narrowed by multicycles of such core-cladding modal couplings, which is of great benefit to the high-resolution sensing applications. Based on such a guiding regime, a high-temperature sensor head is also made and its response to temperature is tested to be of 59.9 pm/ degrees C. PMID- 21643067 TI - Widely-tunable parametric short-wave infrared transmitter for CO2 trace detection. AB - An all-fiber, tunable, short-wave infrared transmitter is demonstrated using efficient four-wave mixing in conventional L and O bands. To realize this source a highly-nonlinear fiber, exhibiting low bend loss over the short-wave infrared spectral band, is employed because of its advantageous properties as a nonlinear mixing medium. The transmitter was subsequently exploited to probe and detect trace levels of carbon dioxide in the 2051-nm spectral region where its beam properties, tunability, narrow linewidth, and stability all coalesce to permit this application. This work indicates this transmitter can serve as a robust source for sensing carbon dioxide and other trace gasses in the short-wave infrared spectral region and should therefore play an important role in future applications. PMID- 21643068 TI - Plastic lab-on-a-chip for fluorescence excitation with integrated organic semiconductor lasers. AB - Laser light excitation of fluorescent markers offers highly sensitive and specific analysis for bio-medical or chemical analysis. To profit from these advantages for applications in the field or at the point-of-care, a plastic lab on-a-chip with integrated organic semiconductor lasers is presented here. First order distributed feedback lasers based on the organic semiconductor tris(8 hydroxyquinoline) aluminum (Alq3) doped with the laser dye 4-dicyanomethylene-2 methyl-6-(p-dimethylaminostyril)-4H-pyrane (DCM), deep ultraviolet induced waveguides, and a nanostructured microfluidic channel are integrated into a poly(methyl methacrylate) (PMMA) substrate. A simple and parallel fabrication process is used comprising thermal imprint, DUV exposure, evaporation of the laser material, and sealing by thermal bonding. The excitation of two fluorescent marker model systems including labeled antibodies with light emitted by integrated lasers is demonstrated. PMID- 21643069 TI - Cryptographic scheme using genetic algorithm and optical responses of periodic structures. AB - This study employed the optical responses of periodic structures, multiple variable functions with sufficient complexity, to develop a cryptographic scheme. The characteristics of structures could be delivered easily with the ciphertext, a series of numbers containing plaintext messages. Two optimization methods utilizing a genetic algorithm were adopted to generate the periodic structure profile as a critical encryption/decryption key. The robustness of methods was further confirmed under various limits. The ciphertext could only be decrypted by referring to the codebook after acquiring the pre-determined optical response. The confidentiality and large capacity of the scheme revealed the enhanced coding strategies here while the success of the scheme was demonstrated with the delivery of an example message. PMID- 21643070 TI - Optofluidic refractive-index sensor in step-index fiber with parallel hollow micro-channel. AB - We present a simple refractive index sensor based on a step-index fiber with a hollow micro-channel running parallel to its core. This channel becomes waveguiding when filled with a liquid of index greater than silica, causing sharp dips to appear in the transmission spectrum at wavelengths where the glass-core mode phase-matches to a mode of the liquid-core. The sensitivity of the dip wavelengths to changes in liquid refractive index is quantified and the results used to study the dynamic flow characteristics of fluids in narrow channels. Potential applications of this fiber microstructure include measuring the optical properties of liquids, refractive index sensing, biophotonics and studies of fluid dynamics on the nanoscale. PMID- 21643071 TI - Short-range order and near-field effects on optical scattering and structural coloration. AB - We have investigated wavelength-dependent light scattering in biomimetic structures with short-range order. Coherent backscattering experiments are performed to measure the transport mean free path over a wide wavelength range. Overall scattering strength is reduced significantly due to short-range order and near-field effects. Our analysis explains why single scattering of light is dominant over multiple scattering in similar biological structures and is responsible for color generation. PMID- 21643072 TI - Coupled 2D Ag nano-resonator chains for enhanced and spatially tailored second harmonic generation. AB - We report results of second harmonic generation calculations performed on Silver coupled 2D-nanoresonators. Coupling is responsible for the creation of resonant modes that can be localized on small portions of the structure or distributed over the whole structure. Different field profiles can be obtained by varying the parameters of the input field (i.e. the wavelength). The second harmonic generation nonlinear process is enhanced by the excitation of coupled surface plasmon polaritons. The emitted field is strongly affected by the linear properties of the structure behaving as a nano antenna. We note that different configurations of the pump field lead to different second harmonic far-field emission patterns. Also, we show that the angular emission of the second harmonic field contains information about the spatial location of the pump field hot spots at different frequencies. Applications to a new class of nano sources for single molecule fluorescence and sensors are proposed. PMID- 21643073 TI - Triple-layer guided-mode resonance Brewster filter consisting of a homogenous layer and coupled gratings with equal refractive index. AB - A triple-layer guided-mode resonance Brewster filter consisting of a homogeneous layer and two identical gratings with their refractive indices equal to that of the homogeneous layer is presented. The spectral properties of this filter are analyzed based on the coupling modulation of two identical binary gratings at Brewster angle for a TM-polarized wave. The grating layer between substrate and homogeneous layers can significantly change the linewidth and resonant mode position, which are due to the asymmetric field distribution inside the grating layers. The tunability of the resonance can be altered on different resonant channels and a practical filter can be obtained in TM2 waveguide mode. Variation of filling factor can alter the field localization in the grating structure and significantly adjust the linewidth of the filter. PMID- 21643074 TI - Bandwidth scalable, coherent transmitter based on the parallel synthesis of multiple spectral slices using optical arbitrary waveform generation. AB - We demonstrate an optical transmitter based on dynamic optical arbitrary waveform generation (OAWG) which is capable of creating high-bandwidth (THz) data waveforms in any modulation format using the parallel synthesis of multiple coherent spectral slices. As an initial demonstration, the transmitter uses only 5.5 GHz of electrical bandwidth and two 10-GHz-wide spectral slices to create 100 ns duration, 20-GHz optical waveforms in various modulation formats including differential phase-shift keying (DPSK), quaternary phase-shift keying (QPSK), and eight phase-shift keying (8PSK) with only changes in software. The experimentally generated waveforms showed clear eye openings and separated constellation points when measured using a real-time digital coherent receiver. Bit-error-rate (BER) performance analysis resulted in a BER < 9.8 * 10(-6) for DPSK and QPSK waveforms. Additionally, we experimentally demonstrate three-slice, 4-ns long waveforms that highlight the bandwidth scalable nature of the optical transmitter. The various generated waveforms show that the key transmitter properties (i.e., packet length, modulation format, data rate, and modulation filter shape) are software definable, and that the optical transmitter is capable of acting as a flexible bandwidth transmitter. PMID- 21643075 TI - Numerically extrapolated discrete layer-peeling algorithm for synthesis of nonuniform fiber Bragg gratings. AB - The discrete layer-peeling algorithm (DLPA) requires to discretize the continuous medium into discrete reflectors to synthesize nonuniform fiber Bragg gratings (FBG), and the discretization step of this discrete model should be sufficiently small for synthesis with high accuracy. However, the discretization step cannot be made arbitrarily small to decrease the discretization error, because the number of multiplications needed with the DLPA is proportional to the inverse square of the layer thickness. We propose a numerically extrapolated time domain DLPA (ETDLPA) to resolve this tradeoff between the numerical accuracy and the computational complexity. The accuracy of the proposed ETDLPA is higher than the conventional time domain DLPA (TDLPA) by an order of magnitude or more, with little computational overhead. To be specific, the computational efficiency of the ETDLPA is achieved through numerical extrapolation, and each addition of the extrapolation depth improves the order of accuracy by one. Therefore, the ETDLPA provides us with computationally more efficient and accurate methodology for the nonuniform FBG synthesis than the TDLPA. PMID- 21643076 TI - Optically driven Archimedes micro-screws for micropump application. AB - Archimedes micro-screws have been fabricated by three-dimensional two-photon polymerization using a Nd:YAG Q-switched microchip laser at 532nm. Due to their small sizes they can be easily manipulated, and made to rotate using low power optical tweezers. Rotation rates up to 40 Hz are obtained with a laser power of 200 mW, i.e. 0.2 Hz/mW. A photo-driven micropump action in a microfluidic channel is demonstrated with a non-optimized flow rate of 6 pL/min. The optofluidic properties of such type of Archimedes micro-screws are quantitatively described by the conservation of momentum that occurs when the laser photons are reflected on the helical micro-screw surface. PMID- 21643077 TI - Real-time terahertz near-field microscope. AB - We report a terahertz near-field microscope with a high dynamic range that can capture images of a 370 x 740 MUm2 area at 35 frames per second. We achieve high spatial resolution (14 MUm corresponding to lambda/30 for a center frequency at 0.7 THz) on a large area by combining two novel techniques: terahertz generation by tilted-pulse-front excitation and electro-optic balanced imaging detection using a thin crystal. To demonstrate the microscope capability, we reveal the field enhancement at the gap position of a dipole antenna after the irradiation of a terahertz pulse. PMID- 21643078 TI - On-chip stimulated Brillouin scattering. AB - We demonstrate on-chip stimulated Brillouin scattering (SBS) in an As2S3 chalcogenide rib waveguide. SBS was characterized in a cm long waveguide with a cross-section 4 MUm x 850 nm using the backscattered signal and pump-probe technique. The measured Brillouin shift and its full-width at half-maximum (FWHM) linewidth were ~7.7 GHz and 34 MHz, respectively. Probe vs. pump power measurements at the Brillouin shift were used to obtain the gain coefficient from an exponential fit. The Brillouin gain coefficient obtained was 0.715 x 10(-9) m/W. A probe gain of 16 dB was obtained for a CW pump power of ~300 mW. PMID- 21643079 TI - Single pixel transmissive and reflective liquid crystal display using broadband cholesteric liquid crystal film. AB - We propose a single mode transflective liquid crystal display (LCD) which is operated as the transmissive and reflective modes in a single pixel without dividing into sub-pixels. The single pixel transflective LCD was composed of the cross-polarized nematic LCD as a light modulator and the broadband cholesteric liquid crystal film (BCLCF) as a half mirror. The BCLCF, simply prepared by the exposure of ultraviolet light to the mixture of the nematic LC and the reactive mesogen with chirality, selectively reflects a certain circular polarization but transmits the orthogonal circular polarization in entire visible light. The electro-optical properties in both transmissive and reflective modes coincide with each other. PMID- 21643080 TI - Temperature-insensitive long-wavelength (lambda ~14 um) Quantum Cascade lasers with low threshold. AB - We demonstrate high-performance, long-wavelength (lambda ~14 um) Quantum Cascade (QC) lasers based on a diagonal optical transition and a "two-phonon-continuum" depletion scheme in which the lower laser level is depopulated by resonant longitudinal optical phonon scattering followed by scattering to a lower energy level continuum. A 2.8 mm long QC laser shows a low threshold current density of 2.0 kA/cm2, a peak output power of ~336 mW, and a slope efficiency of 375 mW/A, all at 300 K, with a high characteristic temperature T0 ~310 K over a wide temperature range from 240 K to 390 K. PMID- 21643081 TI - Optical properties of the crescent and coherent applications. AB - By out-of-particle surface plasmon (SP) excitation in the near infrared range, the influences of key parameters on the basic optical properties of the Au crescent are qualitatively studied from the mode dispersion. Based on the coherent control of SP wave, a crescent pair sensor with the intensified extracted signal and the controllability of sensing is proposed. In addition, the crescent half replaced by Ag functioning as a position detector is also proposed. The particular phase of the detecting light as a detection parameter is used to improve the accuracy of the position detection. PMID- 21643082 TI - Nonlinear oscillator metamaterial model: numerical and experimental verification. AB - We verify numerically and experimentally the accuracy of an analytical model used to derive the effective nonlinear susceptibilities of a varactor-loaded split ring resonator (VLSRR) magnetic medium. For the numerical validation, a nonlinear oscillator model for the effective magnetization of the metamaterial is applied in conjunction with Maxwell equations and the two sets of equations solved numerically in the time-domain. The computed second harmonic generation (SHG) from a slab of a nonlinear material is then compared with the analytical model. The computed SHG is in excellent agreement with that predicted by the analytical model, both in terms of magnitude and spectral characteristics. Moreover, experimental measurements of the power transmitted through a fabricated VLSRR metamaterial at several power levels are also in agreement with the model, illustrating that the effective medium techniques associated with metamaterials can accurately be transitioned to nonlinear systems. PMID- 21643083 TI - Novel real-time homodyne coherent receiver using a feed-forward based carrier extraction scheme for phase modulated signals. AB - We report a novel real-time homodyne coherent receiver based on a DPSK optical electrical-optical (OEO) regenerator used to extract a carrier from carrier-less phase modulated signals based on feed-forward based modulation stripping. The performance of this non-DSP based coherent receiver was evaluated for 10.66 Gbit/s BPSK signals. Self-homodyne coherent detection and homodyne detection with an injection-locked local oscillator laser was demonstrated. The performance was evaluated by measuring the electrical signal-to-noise (SNR) and recording the eye diagrams. Using injection-locking for the LO improves the performance and enables homodyne detection with optical injection-locking to operate with carrier-less BPSK signals without the need for polarization multiplexed pilot-tones. PMID- 21643084 TI - Analysis of acceptable spectral windows of quadratic cascaded nonlinear processes in a periodically poled lithium niobate waveguide. AB - We report a systematic and comparative study of the acceptance bandwidths of two cascaded quadratic nonlinear processes in periodically poled lithium niobate waveguides, namely cascaded second-harmonic generation and difference-frequency generation (cSHG/DFG) and cascaded sum-frequency generation and difference frequency generation (cSFG/DFG). We first theoretically and experimentally study the acceptance bandwidths of both the individual second-harmonic generation (SHG) and sum-frequency generation (SFG) processes in the continuous wave (CW) and pulsed-pump regimes. Our results show that the SHG bandwidth is approximately half that of the SFG process in the CW regime, whereas the SHG acceptance bandwidth can approach the CW SFG bandwidth limit when pulsed-pump is used. As a consequence we conclude that the tuning bandwidths of both cascaded processes should be similar in the pulsed pump regime once the pump pulse bandwidths approach that of SFG (i.e. the cSHG/DFG bandwidth is not limited by the CW SHG bandwidth). We confirm that this is the case experimentally. PMID- 21643085 TI - Suitable combination of noble/ferromagnetic metal multilayers for enhanced magneto-plasmonic biosensing. AB - We present a theoretical and experimental study on the biosensing sensitivity of Au/Co/Au multilayers as transducers of the magneto-optic surface-plasmon resonance (MOSPR) sensor. We demonstrate that the sensing response of these magneto-plasmonic (MP) transducers is a trade-off between the optical absorption and the magneto-optical activity, observing that the MP multilayer with larger MO effect does not provide the best sensing response. We show that it is possible to design highly-sensitive MP transducers able to largely surpass the limit of detection of the conventional surface-plasmon-resonance (SPR) sensor. This was proved comparing the biosensing performance of both sensors for the label-free detection of short DNA chains hybridization. For this purpose, we used and tested a novel label-free biofunctionalization protocol based on polyelectrolytes, which increases the resistance of MP transducers in aqueous environments. PMID- 21643086 TI - Asymmetric transmission for linearly polarized electromagnetic radiation. AB - Metamaterials have shown to support the intriguing phenomenon of asymmetric electromagnetic transmission in the opposite propagation directions, for both circular and linear polarizations. In the present article, we propose a criterion on the relationship among the elements of transmission matrix, which allows asymmetrical transmission for linearly polarized electromagnetic radiation only while the reciprocal transmission for circularly one. Asymmetric hybridized metamaterials are shown to satisfy this criterion. The influence from the rotation of the sample around the radiation propagation direction is discussed. A special structure design is proposed, and its characteristics are analyzed by using numerical simulation. PMID- 21643087 TI - Excess quantum noise in optical parametric chirped-pulse amplification. AB - Noise evolution in an optical parametric chirped-pulse amplifier (OPCPA) differs essentially from that of an optical parametric or a conventional laser amplifier, in that an incoherent pedestal is produced by superfluorescence that can overwhelm the signal under strong saturation. Using a model for the nonlinear dynamics consistent with quantum mechanics, we numerically study the evolution of excess noise in an OPCPA. The observed dynamics explain the macroscopic characteristics seen previously in experiments in the practically important saturation regime. PMID- 21643088 TI - Surface plasmon polariton induced optical amplitude and phase modulation in sub wavelength apertures. AB - We report on the amplitude and phase modulation of picosecond optical pulses, near lambda = 800 nm, transmitted through sub-wavelength rectangular apertures in thin gold films with thicknesses of lambda/10 at per-pulse energies of <0.3 nJ or 9 pJ per aperture. Due to the excitation and strong confinement of surface plasmon polaritons in the apertures, the leading edge of a pulse causes a rapid heating of the electrons and lattice to modulate its falling edge. By comparing cross-correlation frequency resolved optical gating measurements with simulations, the thermal effects responsible for the induced pulse dynamics are identified. PMID- 21643089 TI - Metal-insulator-silicon-insulator-metal waveguides compatible with standard CMOS technology. AB - Metal-insulator-silicon-insulator-metal (MISIM) waveguides are proposed and investigated theoretically. They are hybrid plasmonic waveguides, and light is highly confined to the insulator between the metal and silicon. As compared to previous ones, they are advantageous since they may be realized in a simple way by using current standard CMOS technology and their insulator is easily replaceable without affecting the metal and silicon. First, their structure and fabrication process are explained, both of which are compatible with standard CMOS technology. Then, the characteristics of the single MISIM waveguide whose insulator has its original or an adjusted refractive index are analyzed. The analysis demonstrates that its characteristics are comparable to those of previous hybrid plasmonic waveguides and that they are very effectively tuned by changing the refractive index of the insulator. Finally, the characteristics of the two coupled MISIM waveguides are analyzed. Through the analysis, it is obtained how close or far apart they are for efficient power transfer or low crosstalk. MISIM-waveguide-based devices may play an important role in connecting Si-based photonic and electronic circuits. PMID- 21643090 TI - Modeling of CW Yb-doped fiber lasers with highly nonlinear cavity dynamics. AB - We develop a theoretical framework for modeling of continuous wave Yb-doped fiber lasers with highly nonlinear cavity dynamics. The developed approach has shown good agreement between theoretical predictions and experimental results for particular scheme of Yb-doped laser with large spectral broadening during single round trip. The model is capable to accurately describe main features of the experimentally measured laser outputs such as power efficiency slope, power leakage through fibre Bragg gratings, spectral broadening and spectral shape of generated radiation. PMID- 21643091 TI - Population inversion and low cooperative upconversion in Er-doped silicon-rich silicon nitride waveguide. AB - Single-mode, strip-loaded silicon-rich silicon nitride (SRSN) waveguide with 11 at.% excess Si and 1.7*10(20) cm(-3) Er was fabricated and characterized. By using a 350 nm thick SRSN:Er core layer and a 850 nm wide SiO2 strip, a high core mode overlap of 0.85 and low transmission loss of 2.9 dB/cm is achieved. Population inversion of 0.73-0.75, close to the theoretical maximum, is estimated to have been achieved via 1480 nm resonant pumping, indicating that nearly all doped Er in SRSN are optically active. Analysis of the pump power dependence of Er3+ luminescence intensity and lifetime indicate that the Er cooperative upconversion coefficient in SRSN:Er is as low as 2.1*10(-18) cm3/sec. PMID- 21643092 TI - Deformation of partially pumped active mirrors for high average-power diode pumped solid-state lasers. AB - We discuss the deformation of a partially pumped active mirror amplifier as a free standing disk, as implemented in several laser systems. We rely on the Lucia laser project to experimentally evaluate the analytical and numerical deformation models. PMID- 21643093 TI - Four-dimensional X-ray phase tomography with Talbot interferometry and white synchrotron radiation: dynamic observation of a living worm. AB - X-ray Talbot interferometry is attractive as a method for X-ray phase imaging and phase tomography for objects that weakly absorb X-rays. Because X-ray Talbot interferometry has the advantage that X-rays of a broad energy bandwidth can be used, high-speed X-ray phase imaging is possible with white synchrotron radiation. In this paper, we demonstrate time-resolved three-dimensional observation with X-ray Talbot interferometry (namely, four-dimensional X-ray phase tomography). Differential phase images, from which a phase tomogram was reconstructed, were obtained through the Fourier-transform method, unlike the phase-stepping method that requires several (at least three) moire images to be measured sequentially in order to generate one differential phase image. We demonstrate dynamic observation of a living worm in three dimensions with a time resolution of 0.5 s, visualizing a drastic change in the respiratory tract. PMID- 21643094 TI - Theoretical expressions of the angle-of-arrival variance for optical waves propagating through non-Kolmogorov turbulence. AB - Based on the generalized exponential spectrum for non-Kolmogorov atmospheric turbulence, theoretical expressions of the angle-of-arrival (AOA) variance are derived for plane and spherical optical waves propagating through weak turbulence. Without particular assumption, the new expressions relate the AOA variance to the receiver aperture, finite turbulence inner and outer scales, and the optical wavelength. PMID- 21643095 TI - Fusion of infrared and visual images through region extraction by using multi scale center-surround top-hat transform. AB - Fusion of infrared and visual images is an important research area in image analysis. The purpose of infrared and visual image fusion is to combine the image information of the original images into the final fusion result. So, it is crucial to effectively extract the image information of the original images and reasonably combine them into the final fusion image. To achieve this purpose, an algorithm by using multi scale center-surround top-hat transform through region extraction is proposed in this paper. Firstly, multi scale center-surround top hat transform is discussed and used to extract the multi scale bright and dim image regions of the original images. Secondly, the final extracted image regions for image fusion are constructed from the extracted multi scale bright and dim image regions. Finally, after a base image is calculated from the original images, the final extracted image regions are combined into the base image through a power strategy to form the final fusion result. Because the image information of the original images are well extracted and combined, the proposed algorithm is very effective for image fusion. Comparison experiments have been performed on different image sets, and the results verified the effectiveness of the proposed algorithm. PMID- 21643096 TI - Spectral modeling of channel band shapes in wavelength selective switches. AB - A model for characterizing the spectral response of the passband of Wavelength Selective Switches (WSS) is presented. We demonstrate that, in contrast to the commonly used supergaussian model, the presented model offers a more complete match to measured results, as it is based on the physical operation of the optical system. We also demonstrate that this model is better suited for calculation of WSS channel bandwidths, as well as predicting the final bandwidth of cascaded WSS modules. Finally, we show the utility of this model in predicting channel shapes in flexible bandwidth WSS channel plans. PMID- 21643097 TI - Absorption imaging of ultracold atoms on atom chips. AB - Imaging ultracold atomic gases close to surfaces is an important tool for the detailed analysis of experiments carried out using atom chips. We describe the critical factors that need be considered, especially when the imaging beam is purposely reflected from the surface. In particular we present methods to measure the atom-surface distance, which is a prerequisite for magnetic field imaging and studies of atom surface-interactions. PMID- 21643098 TI - Pedestal cleaning for high laser pulse contrast ratio with a 100 TW class laser system. AB - Laser matter interaction at relativistic intensities using 100 TW class laser systems or higher is becoming more and more widespread. One of the critical issues of such laser systems is to let the laser pulse interact at high intensity with the solid target and avoid any pre-plasma. Thus, a high Laser Pulse Contrast Ratio (LPCR) parameter is of prime importance. We present the LPCR characterization of a high repetition 100 TW class laser system. We demonstrate that the generated Amplified Spontaneous Emission (ASE) degrades the overall LPCR performance. We propose a simple way to clean the pulse after the first amplification stage by introducing a solid state saturable absorber which results in a LPCR improvement to better than 10(10) with only a 30% energy loss at a 10 Hz repetition rate. We finally correlated this cleaning method with experimental results. PMID- 21643099 TI - Controlling total spot power from holographic laser by superimposing a binary phase grating. AB - By superimposing a tunable binary phase grating with a conventional computer generated hologram, the total power of multiple holographic 3D spots can be easily controlled by changing the phase depth of grating with high accuracy to a random power value for real-time optical manipulation without extra power loss. Simulation and experiment results indicate that a resolution of 0.002 can be achieved at a lower time cost for normalized total spot power. PMID- 21643100 TI - Absorption enhancement by matching the cross-section of plasmonic nanowires to the field structure of tightly focused beams. AB - Nanostructured materials, designed for enhanced light absorption, are receiving increased scientific and technological interest. In this paper we propose a physical criterion for designing the cross-sectional shape of plasmonic nanowires for improved absorption of a given tightly focused illumination. The idea is to design a shape which increases the matching between the nanowire plasmon resonance field and the incident field. As examples, we design nanowire shapes for two illumination cases: a tightly focused plane wave and a tightly focused beam containing a line singularity. We show that properly shaped and positioned silver nanowires that occupy a relatively small portion of the beam-waist area can absorb up to 65% of the total power of the incident beam. PMID- 21643101 TI - Enhanced optical transmission through sub-wavelength centered-polygonal hole arrays in silver thin film on silica substrate. AB - We numerically investigated the enhanced optical transmission through sub wavelength centered-polygonal hole arrays (CPHA) in a thin Ag film deposited on the silica substrate. In octagonal and decagonal-CPHAs, we observed new hybrid transmission characteristics that were inherited from both crystalline and quasi crystalline hole arrays. This peculiar nature was attributed to the unique arrangement of CPHAs which can be covered with copies of a single unit cell as in crystalline arrays, and their rotational symmetry as observed in quasi crystalline arrays. Hybrid natures in CPHAs were further investigated in the transmission spectra and Fourier space representations of the arrays. Contributions from the nearest neighbor hole-to-hole distance to enhanced transmission were analyzed in order to quantify the plasmonic contributions from the Air/Ag interface and Silica/Ag interface. We also investigated the impact of layer structure, Air/Ag/Air versus Air/Ag/Silica in the transmissions and found that in CPHAs in Air/Ag/Silica structures, contributions from the Air/Ag interface became dominant in contrast to crystalline hole arrays with lower fold symmetry. PMID- 21643102 TI - Energy flow characteristics of vector X-Waves. AB - The vector form of X-Waves is obtained as a superposition of transverse electric and transverse magnetic polarized field components. It is shown that the signs of all components of the Poynting vector can be locally changed using carefully chosen complex amplitudes of the transverse electric and transverse magnetic polarization components. Negative energy flux density in the longitudinal direction can be observed in a bounded region around the centroid; in this region the local behavior of the wave field is similar to that of wave field with negative energy flow. This peculiar energy flux phenomenon is of essential importance for electromagnetic and optical traps and tweezers, where the location and momenta of micro-and nanoparticles are manipulated by changing the Poynting vector, and in detection of invisibility cloaks. PMID- 21643103 TI - Blind adaptive equalization of polarization-switched QPSK modulation. AB - Coherent detection in combination with digital signal processing has recently enabled significant progress in the capacity of optical communications systems. This improvement has enabled detection of optimum constellations for optical signals in four dimensions. In this paper, we propose and investigate an algorithm for the blind adaptive equalization of one such modulation format: polarization-switched quaternary phase shift keying (PS-QPSK). The proposed algorithm, which includes both blind initialization and adaptation of the equalizer, is found to be insensitive to the input polarization state and demonstrates highly robust convergence in the presence of PDL, DGD and polarization rotation. PMID- 21643104 TI - 3D shape measurement technique for multiple rapidly moving objects. AB - Recently proposed binary defocusing techniques have led to ultrafast speed 3D shape measurement, but they are generally limited to measurement of a single object at a time. Introducing additional gray coded patterns for point-by-point phase unwrapping could permit simultaneous multiple-object measurement. However, when the objects are moving rapidly, the displacement between the first captured pattern and the last can be significant, and pose challenges related to the precisely designed gray codes. This paper presents a new phase unwrapping strategy that combines the conventional spatial phase unwrapping with the gray code to resolve motion related phase unwrapping problems. A system with a speed of 5,000 Hz was developed to verify the performance of the proposed technique. PMID- 21643105 TI - InGaAs-InP avalanche photodiodes with dark current limited by generation recombination. AB - Separate absorption grading charge multiplication avalanche photodiodes (SAGCM APDs) are widely accepted in photon starved optical communication systems due to the presence of large photocurrent gain. In this work, we present a detailed analysis of dark currents of planar-type SAGCM InGaAs-InP APDs with different thicknesses of multiplication layer. The effect of the diffusion process, the generation-recombination process, the tunneling process and the multiplication process on the total leakage current is discussed. A new empirical formula has been established to predict the optimal multiplication layer thickness of SAGCM APDs with dark current limited by generation-recombination at multiplication gain of 8. PMID- 21643106 TI - Design for the optical retardation in broadband zero-order half-wave plates. AB - This paper presents a novel design for broadband zero-order half-wave plates to eliminate the first-order or up to second-order wavelength-dependent birefringent phase retardation (BPR) with 2 or 3 different birefringent materials. The residual BPRs of the plates increase monotonously with the wavelength deviation from a selected wavelength, so the plates are applicable to the broadband light pulses which gather most of the light energy around their central wavelengths. The model chooses the materials by the birefringent dispersion coefficient and evaluates the performances of the plates by the weighted average of the absolute value of residual BPR in order to emphasize the contributions of the incident spectral components whose possess higher energies. PMID- 21643107 TI - Brillouin scattering gain bandwidth reduction down to 3.4MHz. AB - We present a simple method for the stimulated Brillouin scattering (SBS) gain bandwidth reduction in an optical fiber. We were able to reduce the natural bandwidth of 20 MHz to around 3.4 MHz by a superposition of the gain with two losses produced by the same source. This reduced bandwidth can drastically enhance the performance of many different applications which up to now were limited by the minimum of the natural SBS bandwidth. PMID- 21643108 TI - Optical interconnection for a polymeric PLC device using simple positional alignment. AB - This study proposes a simple cost-effective method of optical interconnection between a planar lightwave circuit (PLC) device chip and an optical fiber. It was conducted to minimize and overcome the coupling loss caused by lateral offset which is due to the process tolerance and the dimensional limitation existing between PLC device chips and fiber array blocks with groove structures. A PLC device chip and a fiber array block were simultaneously fabricated in a series of polymer replication processes using the original master. The dimensions (i.e., width and thickness) of the under-clad of the PLC device chip were identical to those of the fiber array block. The PLC device chip and optical fiber were aligned by simple positional control for the vertical direction of the PLC device chip under a particular condition. The insertion loss of the proposed 1 x 2 multimode optical splitter device interconnection was 4.0 dB at 850 nm and the coupling loss was below 0.1 dB compared with single-fiber based active alignment. PMID- 21643109 TI - Phase-controlled superimposed FBGs and their applications in spectral-phase en/decoding. AB - A novel kind of superimposed fiber Bragg gratings (SI-FBGs) named SI-sampled FBGs (SI-SFBGs) is proposed to control the phase relationship among SI sub-gratings by modulating the sampling periods. The realization of such phase-controlled SI SFBGs just needs a single uniform phase mask and sub-micrometer precision moving stage. The success of phase-controll is expected to encourage SI-SFBGs' applications in more sophisticated fields. As a demonstration, their applications in spectral-phase en/decoding are testified by both simulation and experiment. The spectral-phase encoded (SPE) encoders with the longest code-length that FBG based SPE encoders can achieve, i.e., 64-frequency bins, are experimentally fabricated for the first time. The results show the advantages accompanying the SI-SFBGs-based SPE encoders compared with the traditional methods. PMID- 21643110 TI - Optical transmittance degradation in tapered fibers. AB - We investigated the cause of optical transmittance degradation in tapered fibers. Degradation commences immediately after fabrication and it eventually reduces the transmittance to almost zero. It is a major problem that limits applications of tapered fibers. We systematically investigated the effect of the dust-particle density and the humidity on the degradation dynamics. The results clearly show that the degradation is mostly due to dust particles and that it is not related to the humidity. In a dust free environment it is possible to preserve the transmittance with a degradation of less than the noise (+/-0.02) over 1 week. PMID- 21643111 TI - Optofluidic waveguides for reconfigurable photonic systems. AB - We report the development of two liquid waveguide based photonic elements for use in reconfigurable photonic systems. This work demonstrates the ability to couple light from a conventional optical fiber to an adaptable liquid-core/liquid cladding waveguide and back again to an optical fiber(s) enabling us to take advantage of both liquid- and solid-state photonic modalities. We demonstrate and characterize the use of this fiber-in and fiber-out system as either an optical switch or signal attenuator. Microscale flow control enables the adaptive morphology and tunable position of the liquid waveguide yielding an attenuation range of 3.1-10.7 dB, operability over a broad bandwidth spanning the range of wavelengths from visible to telecommunication, and a 1x2 sub-second switching system with a cross-talk as low as 20 dB and maximum coupling efficiency of 3.87 dB. PMID- 21643112 TI - Gain-assisted transformation optics. AB - Loss severely degrades the cloaking effect of the device designed by traditional transformation. In this letter, we propose gain-assisted transformation optics to overcome the loss problem by introducing gain media into a spherical cloak. The gain media, which can amplify the electromagnetic fields, is controlled precisely to compensate the inherent loss in experimental realization of cloaks. We discuss the significance of controlling embedded gain materials in the context of the inverse design mechanism, which allows us to wisely select realizable materials with constant gain and loss along the radius. For practical realizations, isotropic spherical gain-assisted cloak is designed. Full-wave simulations validate the proposed design concept, which can be utilized to alleviate the inevitable loss problem in transformational optical devices. PMID- 21643113 TI - Phase closure retrieval in an infrared-to-visible upconversion interferometer for high resolution astronomical imaging. AB - This paper demonstrates the use of a nonlinear upconversion process to observe an infrared source through a telescope array detecting the interferometric signal in the visible domain. We experimentally demonstrate the possibility to retrieve information on the phase of the object spectrum of an infrared source by using a three-arm upconversion interferometer. We focus our study on the acquisition of phase information of the complex visibility by means of the phase closure technique. In our experimental demonstration, a laboratory binary star with an adjustable photometric ratio is used as a test source. A real time comparison between a standard three-arm interferometer and our new concept using upconversion by sum-frequency generation demonstrates the preservation of phase information which is essential for image reconstruction. PMID- 21643114 TI - Homogeneous optical cloak constructed with uniform layered structures. AB - The prospect of rendering objects invisible has intrigued researchers for centuries. Transformation optics based invisibility cloak design is now bringing this goal from science fictions to reality and has already been demonstrated experimentally in microwave and optical frequencies. However, the majority of the invisibility cloaks reported so far have a spatially varying refractive index which requires complicated design processes. Besides, the size of the hidden object is usually small relative to that of the cloak device. Here we report the experimental realization of a homogenous invisibility cloak with a uniform silicon grating structure. The design strategy eliminates the need for spatial variation of the material index, and in terms of size it allows for a very large obstacle/cloak ratio. A broadband invisibility behavior has been verified at near infrared frequencies, opening up new opportunities for using uniform layered medium to realize invisibility at any frequency ranges, where high-quality dielectrics are available. PMID- 21643115 TI - Add-drop Benes network for scalable optical packet networks. AB - For the scalable optical packet transport networks, an innovative design of add drop Benes network (ADBN) is presented where the cost and energy consumption can be considerably reduced by element savings in the architecture. In a WDM optical packet transport switching node, the ADBNs are interconnected to achieve buffer sharing among multiple ADBNs. A corresponding switch configuration algorithm and architecture rules for the single ADBN and shared ADBN are proposed to mitigate the limited connection capability of the proposed ADBN designs. Switch scalability is verified in consideration of a crosstalk noise performance and element counts. PMID- 21643116 TI - Analysis of light propagation in slotted resonator based systems via coupled-mode theory. AB - Optical devices with a slot configuration offer the distinct feature of strong electric field confinement in a low refractive index region and are, therefore, of considerable interest in many applications. In this work we investigate light propagation in a waveguide-resonator system where the resonators consist of slotted ring cavities. Owing to the presence of curved material interfaces and the vastly different length scales associated with the sub-wavelength sized slots and the waveguide-resonator coupling regions on the one hand, and the spatial extent of the ring on the other hand, this prototypical system provides significant challenges to both direct numerical solvers and semi-analytical approaches. We address these difficulties by modeling the slot resonators via a frequency-domain spatial Coupled-Mode Theory (CMT) approach, and compare its results with a Discontinuous Galerkin Time-Domain (DGTD) solver that is equipped with curvilinear finite elements. In particular, the CMT model is built on the underlying physical properties of the slotted resonators, and turns out to be quite efficient for analyzing the device characteristics. We also discuss the advantages and limitations of the CMT approach by comparing the results with the numerically exact solutions obtained by the DGTD solver. Besides providing considerable physical insight, the CMT model thus forms a convenient basis for the efficient analysis of more complex systems with slotted resonators such as entire arrays of waveguide-coupled resonators and systems with strongly nonlinear optical properties. PMID- 21643117 TI - Preferential gain photonic-crystal fiber for mode stabilization at high average powers. AB - We report on the design and experimental investigation of a preferential gain photonic-crystal fiber with a mode-field diameter of 47 um. This few-mode fiber design confines the doping of Ytterbium-ions just to the center of the core and, therefore, promotes fundamental mode operation. In a chirped-pulse amplification system we extracted up to 303 W of average power from this fiber with a measured M2 value of 1.4. PMID- 21643118 TI - Remarkable decrease in threshold for electrically pumped random ultraviolet lasing from ZnO film by incorporation of Zn2TiO4 nanoparticles. AB - We have comparatively investigated electrically pumped random lasing (RL) actions of two metal-insulator-semiconductor structured devices using pure ZnO and Zn2TiO4-nanoparticle-incorporated ZnO films as the semiconductor components i.e. light-emitting layers, respectively. It is demonstrated that the device using the Zn2TiO4-nanoparticle-incorporated ZnO film as the light-emitting layer exhibits a much smaller threshold current for the electrically pumped RL, which is ascribed to the enhanced multiple light scattering by incorporation of Zn2TiO4 nanoparticles into ZnO film. It is believed that this work provides a strategy for developing low-threshold ZnO-based random lasers. PMID- 21643119 TI - Active cophasing and aligning testbed with segmented mirrors. AB - An active segmented cophasing imaging testbed with a high accurate optical adjustment and control in nanoscale is set up. A two-dimensional dispersed fringe analysis method is used to calculate the piston error between the adjacent segments. Segments alignment of tip-tilt is fulfilled by overlapping diffraction pattern centroids of the individual segment on the focal plane. The cophasing errors are corrected by sensitive micro-displacement actuators under a computer closed-loop control. A FISBA interferometer is introduced to evaluate that a final total residual surface error of about 0.08lambda RMS and the residual piston error of 0.026lambda RMS are achieved (lambda = 633 nm). PMID- 21643120 TI - Optimized electron beam writing strategy for fabricating computer-generated holograms based on an effective medium approach. AB - Recent research revealed that using the effective medium approach to generate arbitrary multi-phase level computer-generated holograms is a promising alternative to the conventional multi-height level approach. Although this method reduces the fabrication effort using one-step binary lithography, the subwavelength patterning process remains a huge challenge, particularly for large scale applications. To reduce the writing time on variable shaped electron beam writing systems, an optimized strategy based on an appropriate reshaping of the binary subwavelength structures is illustrated. This strategy was applied to fabricate a three-phase level CGH in the visible range, showing promising experimental results. PMID- 21643121 TI - Strehl ratios characterizing optical elements designed for presbyopia compensation. AB - We present results of numerical analysis of the Strehl ratio characteristics for the light sword optical element (LSOE). For comparison there were analyzed other optical imaging elements proposed for compensation of presbyopia such as the bifocal lens, the trifocal lens, the stenopeic contact lens, and elements with extended depth of focus (EDOF), such as the logarithmic and quartic axicons. The simulations were based on a human eye's model being a simplified version of the Gullstrand model. The results obtained allow to state that the LSOE exhibits much more uniform characteristics of the Strehl ratio comparing with other known hitherto elements and therefore it could be a promising aid to compensate for the insufficient accommodation range of the human eye. PMID- 21643122 TI - Modulation of spectral intensity, polarization and coherence of a stochastic electromagnetic beam. AB - Analytical formula for the cross-spectral density matrix of a stochastic electromagnetic Gaussian Schell-model (EGSM) beam truncated by a circular phase aperture propagating in free space is derived with the help of a tensor method, which provides a reliable and fast way for studying the propagation and transformation of a truncated EGSM beam. Statistics properties, such as the spectral intensity, the degree of coherence, the degree of polarization and the polarization ellipse of a truncated EGSM beam in free space are studied numerically. The propagation factor of a truncated EGSM beam is also analyzed. Our numerical results show that we can modulate the spectral intensity, the polarization, the coherence and the propagation factor of an EGSM beam by a circular phase aperture. It is found that the phase aperture can be used to shape the beam profile of an EGSM beam and generate electromagnetic partially coherent dark hollow or flat-topped beam, which is useful in some applications, such as optical trapping, material processing, free-space optical communications. PMID- 21643123 TI - Design and fabrication of 3MUm silicon-on-insulator waveguide integrated Ge electro-absorption modulator. AB - We present the design and fabrication of a waveguide-based Ge electro-absorption (EA) modulator integrated with a 3 um silicon-on-isolator (SOI) waveguide. The proposed Ge EA modulator employs a butt-coupled horizontally-oriented p-i-n structure. The optical design achieves a low-loss transition from Ge to Si waveguides. The interaction between the optical mode of the waveguide and the bias induced electric field in the p-i-n structure was maximized to achieve high modulation efficiency. By balancing the trade-offs between the extinction ratio and the insertion loss of the device, an optimal working regime was identified. The measurement results from a fabricated device were used to verify the design. Under a -4Vpp reverse bias, the device demonstrates a total insertion loss (including the transition loss) of 2.7-5.2 dB and an extinction ratio of 4.9-8.2 dB over the wavelength range of 1610-1640 nm. Subtracting the contribution of the transition loss, the Deltaalpha/alpha value for the fabricated device was estimated to be between 2.2 and 3.2 with an electric field around 55 kV/cm. PMID- 21643124 TI - Sub-wavelength bolometers: uncooled platinum wires as infrared sensors. AB - We present characterization results of microscopic platinum wires as bolometers. The wire lengths range from 16 MUm down to 300 nm. Thus they are in many cases significantly smaller in size than the wavelength of the radiation from the 1200 K blackbody source they were exposed to. We observe a steep rise in both responsivity R and detectivity D* with decreasing wire size, reaching R = 3.1*10(4) V/W and D* = 2.7*10(9) cm Hz(1/2)/W at room temperature for a 300*300 nm(2) device. Two significant advantages of such small wires as bolometers are their low power requirement and fast response time. Our numerical estimations suggest response times in the order of nanoseconds for the smallest samples. They could help improve resolution and response of thermal imaging devices, for example. We believe the performance may be further improved by optimizing the design and operating parameters. PMID- 21643125 TI - Enhanced random lasing in ZnO nanocombs assisted by Fabry-Perot resonance. AB - The ultraviolet random lasing behavior of an ensemble of ZnO nanocombs has been demonstrated. It is found that the Fabry-Perot resonance induced by nanocomb geometry can greatly enhance random lasing action with a low threshold condition. Besides, the emission spectra exhibit few sharp lasing peaks with a full width at half maximum (FWHM) of less than 0.3 nm and a narrow background emission with a FWHM of about 5 nm. Cathodoluminescence mapping images are utilized to analyze the Fabry-Perot resonance phenomenon. The resonant effect on the lasing system is further confirmed by nanocombs with different resonant cavity lengths. The unique lasing behavior induced by the simultaneous occurrence of Fabry-Perot resonance and random laser action shown here may open up a new possibility for the creation of highly efficient light emitting devices. PMID- 21643126 TI - Silicon-based optical leaky wave antenna with narrow beam radiation. AB - We propose a design of a dielectric (silicon nitride) optical leaky wave antenna (OLWA) with periodic semiconductor (silicon) corrugations, capable of producing narrow beam radiation. The optical antenna radiates a narrow beam because a leaky wave (LW) with low attenuation constant is excited at one end of the corrugated dielectric waveguide. We show that pointing angle, beam-width, and operational frequency are all related to the LW complex wavenumber, whose value depends on the amount of silicon perturbations in the waveguide. In this paper, the propagation constant and the attenuation coefficient of the LW in the periodic structure are extracted from full-wave simulations. The far-field radiation patterns in both glass and air environments predicted by LW theory agree well with the ones obtained by full-wave simulations. We achieve a directive radiation pattern in glass environment with about 17.5 dB directivity and 1.05 degree beam width at the operative free space wavelength of 1.55 MUm, pointing at a direction orthogonal to the waveguide (broadside direction). We also show that the use of semiconductor corrugations facilitate electronic tuning of the radiation pattern via carrier injection. PMID- 21643127 TI - Cell-based hardware architecture for full-parallel generation algorithm of digital holograms. AB - This paper proposes a new hardware architecture to speed-up the digital hologram calculation by parallel computation. To realize it, we modify the computer generated hologram (CGH) equation and propose a cell-based very large scale integrated circuit architecture. We induce a new equation to calculate the horizontal or vertical hologram pixel values in parallel, after finding the calculation regularity in the horizontal or vertical direction from the basic CGH equation. We also propose the architecture of the computer-generated hologram cell consisting of an initial parameter calculator and update-phase calculators based on the equation, and then implement them in hardware. Modifying the equation could simplify the hardware, and approximating the cosine function could optimize the hardware. In addition, we show the hardware architecture to parallelize the calculation in the horizontal direction by extending computer generated holograms. In the experiments, we analyze hardware resource usage and the performance-capability characteristics of the look-up table used in the computer-generated hologram cell. These analyses make it possible to select the amount of hardware to the precision of the results. Here, we used the platform from our previous work for the computer-generated hologram kernel and the structure of the processor. PMID- 21643128 TI - Chaos synchronization in mutually coupled semiconductor lasers with asymmetrical bias currents. AB - We experimentally and numerically investigated the chaos synchronization characteristics of mutually coupled semiconductor lasers (MCSLs) with asymmetrical bias currents. Experimental results show that, asymmetrical bias current level of two MCSLs has obvious influence on chaos synchronization between them, and stable leader-laggard chaos synchronization can be realized under relatively large asymmetrical bias current levels. Moreover, the influences of frequency detuning and mutually coupling strength between the two lasers on chaos synchronization performance have also been discussed. Theoretical simulations basically conform to our experimental observations. PMID- 21643129 TI - Generation of Cerenkov radiation at 850 nm in higher-order-mode fiber. AB - We demonstrate generation of Cerenkov radiation at 850 nm in a higher-order-mode (HOM) fiber. The LP02 mode in this solid, silica-based fiber has anomalous dispersion from 690 nm to 810 nm. Cerenkov radiation with 3 nJ pulse energy is generated in this module, exhibiting 60% energy conversion efficiency from the input. The HOM fiber provides a valuable fiber platform for nonlinear wavelength conversion with pulse energies in-between index-guided silica-core photonic crystal fibers and air-core photonic bandgap fibers. PMID- 21643130 TI - Improved arrayed-waveguide-grating layout avoiding systematic phase errors. AB - We present a detailed description of an improved arrayed-waveguide-grating (AWG) layout for both, low and high diffraction orders. The novel layout presents identical bends across the entire array; in this way systematic phase errors arising from different bends that are inherent to conventional AWG designs are completely eliminated. In addition, for high-order AWGs our design results in more than 50% reduction of the occupied area on the wafer. We present an experimental characterization of a low-order device fabricated according to this geometry. The device has a resolution of 5.5 nm, low intrinsic losses (< 2 dB) in the wavelength region of interest for the application, and is polarization insensitive over a wide spectral range of 215 nm. PMID- 21643131 TI - Dynamics and all-optical control of solitons at the interface of optical superlattices with spatially modulated nonlinearity. AB - We find the existence of two kinds of solitons at the interface of optical superlattices with both spatially modulated nonlinearity and linear refraction index. The first kind of solitons can either drift across the lattice, or deflect to the uniform nonlinear medium. The dynamics of such solitons mainly depends on their powers. The other kind of solitons can stably propagate along the surface, and can be controlled by additional Gaussian beams. In addition, we demonstrate the input-angle-dependent reflection, trapping, and refraction with nearly no losses by launching sech-shaped solitons. PMID- 21643132 TI - Instantaneous lineshape analysis of Fourier domain mode-locked lasers. AB - We present a theoretical and experimental analysis of the instantaneous lineshape of Fourier domain mode-locked (FDML) lasers, yielding good agreement. The simulations are performed employing a recently introduced model for FDML operation. Linewidths around 10 GHz are found, which is significantly below the sweep filter bandwidth. The effect of detuning between the sweep filter drive frequency and cavity roundtrip time is studied revealing features that cannot be resolved in the experiment, and shifting of the instantaneous power spectrum against the sweep filter center frequency is analyzed. We show that, in contrast to most other semiconductor based lasers, the instantaneous linewidth is governed neither by external noise sources nor by amplified spontaneous emission, but it is directly determined by the complex FDML dynamics. PMID- 21643133 TI - Transmission of 107-Gb/s mode and polarization multiplexed CO-OFDM signal over a two-mode fiber. AB - In addition to the dimensions of time, frequency, complex constellation, and polarization, spatial mode can be the fifth dimension to be explored for modulation and multiplexing in optical fiber communications. In this paper, we demonstrate successful transmission of 107-Gb/s dual-mode and dual-polarization coherent optical orthogonal frequency-division multiplexing (CO-OFDM) over a 4.5 km two-mode fiber. A mechanically-induced LP01/LP11 mode converter is used as the mode selective element in a spatial-mode multiplexed system. PMID- 21643134 TI - A periodically coupled plasmon nanostructure for refractive index sensing. AB - We present unique characteristics of subwavelength surface plasmon polaritons in a periodically coupled nanowell structure. The nanowell structure offers high quality internal surface plasmon resonance for sensing applications. Calculated FWHM of the transmission peak is 6 nm and the optical transmission is close to 100% at the resonant wavelength of 815.8 nm. The highly concentrated polaritons in the nanowell are sensitive to surface changes providing a sensitivity of 4800% RIU(-1) for optical sensing applications. PMID- 21643135 TI - Investigation of strain sensing effect in modified single-defect photonic crystal nanocavity. AB - This paper reports the theoretical and experimental investigations on the strain sensing effect of a two dimensions (2D) photonic crystal (PhC) nanocavity resonator. By using the finite element method (FEM) and finite difference time domain (FDTD) simulations, the strain sensitivity of a high quality factor PhC nanocavity was calculated. Linear relationships between the applied strain and the shift in the resonant wavelength of the cavity were obtained. A single-defect silicon (Si) PhC cavity was fabricated, and measurements of the strain sensitivity were performed. Good agreement between the experimental and simulation results was observed. PMID- 21643136 TI - Measurement of the transmission of the atmosphere from 0.2 to 2 THz. AB - The attenuation of electromagnetic wave propagation in the clear atmosphere from low frequencies up to 2 THz is mainly caused by water vapor. Although there have been many numerical simulations and excellent early sub-mm and far-infrared measurements of this attenuation, there has remained controversy about the background absorption in the most promising windows of transparency below 1 THz. Here, we report an accurate terahertz time-domain spectroscopy (THz-TDS) characterization of water vapor from 0.2 to 2 THz. Our results agree with the previous predicted and measured attenuations for the weak water lines, but show more attenuation for the relatively transparent windows between these lines. PMID- 21643137 TI - SSBI mitigation at 60GHz OFDM-ROF system based on optimization of training sequence. AB - We have theoretically and experimentally investigated the effect of the interference between subcarrier-signal beat interference (SSBI) in 60 GHz orthogonal frequency division multiplexing - radio-over-fiber (OFDM-ROF) system. In order to reduce the influence of SSBI, we compared four kinds of OFDM frames with different training sequences as all-real, all-complex, complex-zero and real zero training. The experimental results show the power penalty of all-real, all complex, complex-zero and real-zero training is 2.5, 5.5, 4 and 1dB at BER of 1x10(-3) after 20 km standard single mode fiber (SMF) transmission, respectively. The real-zero training OFDM frame with interleave structure and lower modulation order signal suffered from the least SSBI shows the best performance. PMID- 21643138 TI - Optical tweezers based active microrheology of sodium polystyrene sulfonate (NaPSS). AB - We used oscillatory optical tweezers to investigate the microrheological properties of Sodium polystyrene sulfonate (NaPSS; Mw = 70 kDa) polymer solutions with different concentrations from 0.001 mM to 10 mM in terms of elastic modulus G'(omega) and loss modulus G"(omega) as a function of angular frequency (omega) in the range of 6 rad/s to 6000 rad/s. The viscoelastic properties (including zero-shear-rate viscosity, crossing frequency and transition frequency) as a function of polymer concentration, deduced from our primary data, reveal the subtle structural changes in the polymer solutions as the polymer concentration increases from dilute to semi-dilute regimes, passing through the critical micelle formation concentration and the polymer overlapping concentration. The experimental results are consistent with the Maxwell model in some regime, and with the Rouse model in other, indicating the transient network character and the micelles formation in different regimes. PMID- 21643139 TI - Surface plasmon polariton absorption modulator. AB - An electrically controlled ultra-compact surface plasmon polariton absorption modulator (SPPAM) is proposed. The device can be as small as a few micrometers depending on the required extinction ratio and the acceptable loss. The device allows for operation far beyond 100 Gbit/s, being only limited by RC time constants. The absorption modulator comprises a stack of metal/insulator/metal oxide/metal layers, which support a strongly confined asymmetric surface plasmon polariton (SPP) in the 1.55 MUm telecommunication wavelength window. Absorption modulation is achieved by electrically modulating the free carrier density in the intermediate metal-oxide layer. The concept is supported by proof-of-principle experiments. PMID- 21643140 TI - Investigation of ultrashort pulse laser ablation of solid targets by measuring the ablation-generated momentum using a torsion pendulum. AB - 50 fs - 12 ps laser pulses are employed to ablate aluminum, copper, iron, and graphite targets. The ablation-generated momentum is measured with a torsion pendulum. Corresponding time-resolved shadowgraphic measurements show that the ablation process at the optimal laser fluence achieving the maximal momentum is primarily dominated by the photomechanical mechanism. When laser pulses with specific laser fluence are used and the pulse duration is tuned from 50 fs to 12 ps, the generated momentum firstly increases and then remains almost constant, which could be attributed to the change of the ablation mechanism involved from atomization to phase explosion. The investigation of the ablation-generated momentum also reveals a nonlinear momentum-energy conversion scaling law, namely, as the pulse energy increases, the momentum obtained by the target increases nonlinearly. This may be caused by the effective reduction of the dissipated energy into the surrounding of the ablation zone as the pulse energy increases, which indicates that for femtosecond laser the dissipated energy into the surrounding target is still significant. PMID- 21643141 TI - Path-length resolved reflectance in tendon and muscle. AB - Optical diffuse reflectance in fibrous tissues depends on measurement angles in relation to fiber orientation. In this study, path-length resolved optical reflectance was measured in tendon and skeletal muscle samples using a low coherence Mach-Zehnder interferometer. The results show that the angular dependency in reflectance was eliminated in tendon tissue when representing reflectance as a function of mean path-length. Our analysis indicated that this observation can be understood in the frame work of anisotropic diffuse theory. However the same phenomenon was not observed in muscles, suggesting involvement of additional scattering mechanisms. PMID- 21643142 TI - Silicon-based horizontal nanoplasmonic slot waveguides for on-chip integration. AB - Horizontal metal/insulator/Si/insulator/metal nanoplasmonic slot waveguide (PWG), which is inserted in a conventional Si wire waveguide, is fabricated using the standard Si-CMOS technology. A thin insulator between the metal and the Si core plays a key role: it not only increases the propagation distance as the theoretical prediction, but also prevents metal diffusion and/or metal-Si reaction. Cu-PWGs with the Si core width of ~134-21 nm and ~12-nm-thick SiO2 on each side exhibit a relatively low propagation loss of ~0.37-0.63 dB/um around the telecommunication wavelength of 1550 nm, which is ~2.6 times smaller than the Al-counterparts. A simple tapered coupler can provide an effective coupling between the PWG and the conventional Si wire waveguide. The coupling efficiency as high as ~0.1-0.4 dB per facet is measured. The PWG allows a sharp bending. The pure bending loss of a Cu-PWG direct 90 degrees bend is measured to be ~0.6-1.0 dB. These results indicate the potential for seamless integration of various functional nanoplasmonic devices in existing Si electronic photonic integrated circuits (Si-EPICs). PMID- 21643143 TI - Optical cavity characterization in nanowires via self-generated broad-band emission. AB - Broadband white light is of great spectroscopic value and would be a powerful tool for nanoscale spectroscopy, however, generation and direction of white light on this length scale remains challenging. Here, we demonstrate the generation of broadband white light in sub-wavelength diameter Gallium Nitride (GaN) wires by coincident one- and two-photon absorption mediated via defect states. This generation of broadband, "white" light enables single-nanowire interferometric measurements of the nanowires themselves via analysis of the Fabry-Perot fringes that overlay the entirety of the emission spectrum. The quality factor and finesse of individual nanowire cavities were measured and calculated to be 186 +/ 88 and 3.05 +/-0.6 respectively, averaged over 20 individual wires. This work presents a new, simple approach for the generation and direction of broad band white light at sub-diffraction limit length scales, ideal for translating classical white light spectroscopies to higher spatial resolutions then previously achieved. PMID- 21643144 TI - Manipulating the plasmon-induced transparency in terahertz metamaterials. AB - Coupling between superradiant and subradiant mode resonators in a metamaterial unit cell plays an important role in observing the sharp transparency peak due to destructive interference between the resonators. This effect is enhanced as the resonators are brought closer to each other in a conventional planar arrangement. We present a novel coupling scheme of planar terahertz metamaterial to tune the plasmon-induced transparency peak by physically varying the distance between the superradiant and the subradiant resonators in such a way that the transparency peak begins to disappear as the coupled resonators are brought closer than a critical separation distance. The effect is attributed to the disappearance of the resonant behavior of the subradiant resonator in a closely coupled regime. The simple planar design presented here demonstrates a scheme to manipulate the electromagnetically induced transparency-like behavior in terahertz metamaterials and this could lead to the development of unique slow light devices for terahertz applications. PMID- 21643145 TI - Optical-parametric-amplification imaging of complex objects. AB - We used ultrafast Fourier-plane optical-parametric-amplification (OPA) imaging to simultaneously image, wavelength-shift, and amplify complex two-dimensional objects with spatial features from 1.1 to 10.1 line pairs/millimeter (lp/mm) in the vertical dimension and from 2.0 to 16.0 lp/mm in the horizontal dimension, corresponding to a two-dimensional space-bandwidth product (SBP) of ~46,000. This represents an increase in image complexity over previous analogous OPA imaging systems by over three orders of magnitude. We observe both wavelength-shifting the image from 930 nm to a wavelength of 700 nm and image amplification by two orders of magnitude. Our wavelength-shifted image has a SBP of ~30,000. PMID- 21643146 TI - Modeling and analysis of localized biosensing and index sensing by introducing effective phase shift in microfiber Bragg grating (uFBG). AB - We report a novel micro-fiber Bragg grating (uFBG) sensor that takes advantage of the degeneracy of stop-band and rapid emergence of spectral modes when an effective phase shift occurs. The phase shift can be enabled by a range of perturbations in a central segment of the grating, including monolayer immobilization of bio-molecules or change in refractive index in the surrounding, thereby constituting the possibility of a highly sensitive sensor with the merit of scalable performance. The use of uFBG ensures strong evanescent field coupling to the surrounding in order to maximize signal transduction. Simulation results indicate very favorable sensor signal characteristics such as large wavelength shift and sharp reflection dips. A general relation between the peak position within the stop-band and the amount of effective phase shift is also provided, and may generally serve as helpful guideline for FBG sensor design. A typical uFBG sensor device may detect surface protein/DNA adsorption with limit-of detection (LOD) as low as 3.3 pg.mm(-2) for surface mass density and 51.8 fg for total mass. For refractive index (RI) sensing, the LOD is 2.5*10(-6) refractive index unit (RIU). PMID- 21643147 TI - Comparison of numerical methods in near-field computation for metallic nanoparticles. AB - Four widely used electromagnetic field solvers are applied to the problem of scattering by a spherical or spheroidal silver nanoparticle in glass. The solvers are tested in a frequency range where the imaginary part of the scatterer refractive index is relatively large. The scattering efficiencies and near-field results obtained by the different methods are compared to each other, as well as to recent experiments on laser-induced shape transformation of silver nanoparticles in glass. PMID- 21643148 TI - A room-temperature semiconductor spaser operating near 1.5 MUm. AB - Room temperature spasing of surface plasmon polaritons at 1.46 MUm wavelength has been demonstrated by sandwiching a gold-film plasmonic waveguide between optically pumped InGaAs quantum-well gain media. The spaser exhibits gain narrowing, the expected transverse-magnetic polarization, and mirror feedback provided by cleaved facets in a 1-mm long cavity fabricated with a flip-chip approach. The 1.06-MUm pump-threshold of ~60 kW/cm2 is in good agreement with calculations. The architecture is readily adaptable to all-electrical operation on an integrated microchip. PMID- 21643149 TI - Nearly perfect absorption in intrinsically low-loss grating structures. AB - The feature of enhanced absorption in two-layered grating structures is theoretically investigated. The underlying structures make the most use of resonance mechanism to achieve a nearly perfect absorption in an intrinsically low-loss medium. For standalone gratings, the maximum absorption efficiency is shown to be 50%, which is attributed to the coupling of short range (bonding) or long range (antibonding) surface plasmons with cavity resonances. By attaching a dielectric slab on top or bottom to the metallic grating, the maximum absorption efficiency can be raised to nearly 100%. The presence of guided waves in the dielectric slab causes the strong concentration of fields and reinforces the absorption to its extreme value. The efficient absorption mechanism is illustrated with the pattern of resonance fields and the distribution of power loss density. A phenomenological theory is also used to characterize the absorption anomaly in terms of complex pole and zero. PMID- 21643150 TI - Chirped-pulse terahertz spectroscopy for broadband trace gas sensing. AB - We report the first demonstration of a broadband trace gas sensor based on chirp pulse terahertz spectroscopy. The advent of newly developed solid state sources and sensitive heterodyne detectors for the terahertz frequency range have made it possible to generate and detect precise arbitrary waveforms at THz frequencies with ultra-low phase noise. In order to maximize sensitivity, the sample gas is first polarized using sub-MUs chirped THz pulses and the free inductive decays (FIDs) are then detected using a heterodyne receiver. This approach allows for a rapid broadband multi-component sensing with low parts in 10(9) (ppb) sensitivities and spectral frequency accuracy of <20 kHz in real-time. Such a system can be configured into a portable, easy to use, and relatively inexpensive sensing platform. PMID- 21643151 TI - Fabrication of optical mosaic gratings: a self-referencing alignment method. AB - We propose and demonstrate a self-referencing alignment technique to conveniently enlarge fabricated grating area. The latent image gratings are used as the reference objects to align (adjust and lock) the attitude and position of the substrate relative to the exposure beams between and during consecutive exposures. The adjustment system and the fringe-locking system are combined into the exposure system, eliminating the drift errors between them and making the whole system low-cost and compact. For the fabricated 1 * 4 mosaics of 50 * (30 + 30 + 30 + 30) mm(2) area and 1 * 2 mosaics of 90 * (80 + 80) mm(2) area, the typical peak-valley -1st-order wavefront errors measured by a 100-mm-diameter interferometer are not more than 0.06 lambda and 0.09 lambda, respectively. PMID- 21643152 TI - Low noise chirped pulse mode-locked laser using an intra-cavity Fabry-Perot etalon. AB - This work presents an extensive investigation of the performance characteristics of a semiconductor-based Theta cavity design laser with an intra-cavity Fabry Perot etalon operating at 100 MHz repetition rate. The Theta laser being an external cavity harmonically mode-locked semiconductor laser exhibits supermode noise that impairs its performance. A fiberized Fabry-Perot periodic filter inserted within the Theta laser cavity mitigates the contribution of the supermode noise to the pulse-to-pulse energy variance by 20 times. The laser has both a compressed output with picosecond pulse duration and a uniform intensity quasi-CW linearly chirped pulse output with 10 nm bandwidth. Long-term stability is attained by referencing the cavity length to the etalon using an intra-cavity Hansch-Couillaud locking scheme. PMID- 21643153 TI - Electro-optical Pockels scattering from a single nanocrystal. AB - The electro-optical Pockels response from a single non-centrosymmetric nanocrystal is reported. High sensitivity to the weak electric-field dependent nonlinear scattering is achieved through a dedicated imaging interferometric microscope and the linear dependence of electro-optical signal upon the applied field is checked. Using different incident light polarization states, a priori random spatial orientation of the crystal can be inferred. The electro-optical response from a nanocrystal provides local subwavelength sensor of quasi-static electric fields with potential applications in physics and biology. It also leads to a new sub-wavelength microscopy towards the nanoscale investigation of interesting phenomena such as nanoferroelectricity. PMID- 21643154 TI - Optical cryptography topology based on a three-dimensional particle-like distribution and diffractive imaging. AB - In recent years, coherent diffractive imaging has been considered as a promising alternative for information retrieval instead of conventional interference methods. Coherent diffractive imaging using the X-ray light source has opened up a new research perspective for the measurement of non-crystalline and biological specimens, and can achieve unprecedentedly high resolutions. In this paper, we show how a three-dimensional (3D) particle-like distribution and coherent diffractive imaging can be applied for a study of optical cryptography. An optical multiple-random-phase-mask encoding approach is used, and the plaintext is considered as a series of particles distributed in a 3D space. A topology concept is also introduced into the proposed optical cryptosystem. During image decryption, a retrieval algorithm is developed to extract the plaintext from the ciphertexts. In addition, security and advantages of the proposed optical cryptography topology are also analyzed. PMID- 21643155 TI - Electrostatic actuation of silicon optomechanical resonators. AB - Cavity optomechanical systems offer one of the most sensitive methods for detecting mechanical motion using shifts in the optical resonance frequency of the optomechanical resonator. Presently, these systems are used for measuring mechanical thermal noise displacement or mechanical motion actuated by optical forces. Electrostatic capacitive actuation and detection have been shown previously for silicon micro electro mechanical resonators for application in filters and oscillators. Here, we demonstrate monolithic integration of electrostatic capacitive actuation with optical sensing using silicon optomechanical disk resonators and waveguides. The electrically excited mechanical motion is observed as an optical intensity modulation when the input electrical signal is at a frequency of 235 MHz corresponding to the radial vibrational mode of the silicon microdisk. PMID- 21643157 TI - Evidence of surface plasmon resonance in ultrafast laser-induced ripples. AB - The sensitivity of grating-coupled Surface Plasmon Polaritons (SPPs) on metallic surface has been exploited to investigate the correlation between ripples formation under ultrashort laser exposure and SPPs generation conditions. Systematic examination of coupling of single ultrashort laser pulse on gratings with appropriate periods ranging from 440 nm to 800 nm has been performed. Our approach reveals that a surface plasmon is excited only for an appropriate grating period, the nickel sample exhibits fine ripples pattern, evidencing the plasmonic nature of ripples generation. We propose a systematic investigation supported by a comprehensive study on the obtained modulation of such a coupling efficiency by means of a phenomenological Drude-Lorentz model which captures possible optical properties modification under femtosecond irradiation. PMID- 21643156 TI - Pure optical photoacoustic microscopy. AB - The concept of pure optical photoacoustic microscopy(POPAM) was proposed based on optical rastering of a focused excitation beam and optically sensing the photoacoustic signal using a microring resonator fabricated by a nanoimprinting technique. After the refinements of the microring's working wavelength and in the resonator structure and mold fabrication, an ultrahigh Q factor of 3.0*10(5) was achieved which provided high sensitivity with a noise equivalent detectable pressure(NEDP) value of 29 Pa. This NEDP is much lower than the hundreds of Pascals achieved with existing optical resonant structures such as etalons, fiber gratings and dielectric multilayer interference filters available for acoustic measurement. The featured high sensitivity allowed the microring resonator to detect the weak photoacoustic signals from micro- or submicroscale objects. The inherent superbroad bandwidth of the optical microring resonator combined with an optically focused scanning beam provided POPAM with high resolution in the axial as well as both lateral directions while the axial resolution of conventional photoacoustic microscopy (PAM) suffers from the limited bandwidth of PZT detectors. Furthermore, the broadband microring resonator showed similar sensitivity to that of our most sensitive PZT detector. The current POPAM system provides a lateral resolution of 5 MUm and an axial resolution of 8 MUm, comparable to that achieved by optical microscopy while presenting the unique contrast of optical absorption and functional information complementing other optical modalities. The 3D structure of microvasculature, including capillary networks, and even individual red blood cells have been discerned successfully in the proof-of-concept experiments on mouse bladders ex vivo and mouse ears in vivo. The potential of approximately GHz bandwidth of the microring resonator also might allow much higher resolution than shown here in microscopy of optical absorption and acoustic propagation properties at depths in unfrozen tissue specimens or thicker tissue sections, which is not now imageable with current optical or acoustic microscopes of comparable resolution. PMID- 21643158 TI - Nonlinear penalties in long-haul optical networks employing dynamic transponders. AB - We report for the first time, the impact of cross phase modulation in WDM optical transport networks employing dynamic 28 Gbaud PM-mQAM transponders (m = 4, 16, 64, 256). We demonstrate that if the order of QAM is adjusted to maximize the capacity of a given route, there may be a significant degradation in the transmission performance of existing traffic for a given dynamic network architecture. We further report that such degradations are correlated to the accumulated peak-to-average power ratio of the added traffic along a given path, and that managing this ratio through pre-distortion reduces the impact of adjusting the constellation size of neighboring channels. PMID- 21643159 TI - Polarization independent broadband reflectors based on cross-stacked gratings. AB - We report here a broadband reflector based on a two cross-stacked grating structure. This type of broadband reflector is polarization-independent, with ~100% reflectance over a designed spectral range of 1.4 to 1.6 MUm. The reflection phase differences between TE and TM polarizations remain almost a constant value of 1.2pi over the same high reflection spectral range. The reflector performance tolerance was also investigated by varying the grating structure parameters. Two types of Fabry-Perot cavities can be configured based on two cross-stacked grating structures, for both polarization independent and polarization dependent resonance cavity mode control. All these characteristics associated with the cross-stacked grating reflectors enable a new type of resonant cavity or wave plate design for a large range of photonic applications. PMID- 21643160 TI - Holographic mode-selective launch for bandwidth enhancement in multimode fiber. AB - With rapidly growing bandwidth demands in Local Area Networks, it is imperative to support next generation speeds beyond 40 Gbit/s. Various holographic optimization techniques using spatial light modulators have recently been explored for adaptive channel impulse response improvement of MMF links. Most of these experiments are algorithmic-oriented. In this paper, a set of lenses and a spatial light modulator, acting as a binary amplitude filter, played the pivotal role in generating the input modal electric field into a graded-index MMF, rather than algorithms. By using a priori theoretical information to generate the incident modal electric field at the MMF, the bandwidth was increased by up to 3.4 times. PMID- 21643161 TI - Precession optomechanics. AB - We propose a light-structure interaction that utilizes circularly polarized light to deform a slightly bent waveguide. The mechanism allows for flipping the direction of deformation upon changing the binary polarization state of light from -h to +h. PMID- 21643162 TI - Hollow waveguide photomixing for quantum cascade laser heterodyne spectro radiometry. AB - An integrated optic approach, using hollow waveguides, has been evaluated for a compact, rugged, high efficiency heterodyne optical mixing circuit in the middle infrared. The approach has involved the creation of hollow waveguides and alignment features for a beam combiner component in a glass-ceramic substrate. The performance of the integrated beam combiner was tested as part of a full laser heterodyne spectro-radiometer in which a quantum cascade laser local oscillator emitting at 9.7 um was mixed with incoherent radiation. The performance has been evaluated with both cryogenically-cooled and peltier-cooled photomixers demonstrating consistent detection limits of two and five times the shot noise limit, respectively. The hollow waveguide mixer has also shown advantages in temporal stability, laser spatial mode cleansing, and reduced sensitivity to optical feedback. PMID- 21643163 TI - Calculation for computer generated hologram using ray-sampling plane. AB - We introduce a new algorithm for calculating computer generated hologram (CGH) using ray-sampling (RS) plane. RS plane is set at near the object and the light rays emitted by the object are sampled at the plane. Then the light-rays are transformed into the wavefront with using the Fourier transforms. The wavefront on the CGH plane is calculated by wavefront propagation simulation from RS plane to CGH plane. The proposed method enables to reproduce high resolution image for deep 3D scene with angular reflection properties such as gloss appearance. PMID- 21643164 TI - Compact cryogenic self-aligning fiber-to-detector coupling with losses below one percent. AB - We present a compact packaging technique for coupling light from a single-mode telecommunication fiber to cryogenic single-photon sensitive devices. Our single photon detectors are superconducting transition-edge sensors (TESs) with a collection area only a factor of a few larger than the area of the fiber core which presents significant challenges to low-loss fiber-to-detector coupling. The coupling method presented here has low loss, cryogenic compatibility, easy and reproducible assembly and low component cost. The system efficiency of the packaged single-photon counting detectors is verified by the "triplet method" of power-source calibration along with the "multiple attenuator" method that produces a calibrated single-photon flux. These calibration techniques, when used in combination with through-wafer imaging and fiber back-reflection measurements, give us confidence that we have achieved coupling losses below 1% for all devices packaged according to the self-alignment method presented in this paper. PMID- 21643165 TI - All-optical OFDM transmission of 7 x 5-Gb/s data over 84-km standard single-mode fiber without dispersion compensation and time gating using a photonic-integrated optical DFT device. AB - We implement dispersion-tolerant and time-gating-free all-optical OFDM transmission using a photonic-integrated discrete Fourier transform (DFT) device. We show that 35-Gb/s OFDM data having near-unity spectral efficiency can be transmitted all-optically with 1-dB dispersion margin of ~1000 ps/nm. The passive optical DFT circuit is implemented using multi-mode interference (MMI) couplers on a high index-contrast silica integrated-optic platform. We also propose a photonic DFT circuit based on an NxN MMI device capable of simultaneous channelization of OFDM signals into N subcarriers. PMID- 21643166 TI - Filamentation in air with ultrashort mid-infrared pulses. AB - We theoretically investigate filamentation of ultrashort laser pulses in air in the mid-infrared regime under conditions in which the group-velocity dispersion (GVD) is anomalous. When a high-power, ultra-short mid-infrared laser beam centered at 3.1-MUm forms a filament, a spatial solitary wave is stabilized by the plasma formation and propagates several times its diffraction length. Compared with temporal self-compression in gases due to plasma formation and pulse splitting in the normal-GVD regime, the minimum achievable pulse duration (~70 fs) is limited by the bandwidth of the anomalous-GVD region in air. For the relatively high powers, multiple pulse splitting due to the plasma effect and shock formation is observed, which is similar to that which occurs in solids. Our simulations show that the energy reservoir also plays a critical role for longer propagation of the air filament in the anomalous-GVD regime. PMID- 21643167 TI - Suspended core subwavelength fibers: towards practical designs for low-loss terahertz guidance. AB - In this work we report two designs of subwavelength fibers packaged for practical terahertz wave guiding. We describe fabrication, modeling and characterization of microstructured polymer fibers featuring a subwavelength-size core suspended in the middle of a large porous outer cladding. This design allows convenient handling of the subwavelength fibers without distorting their modal profile. Additionally, the air-tight porous cladding serves as a natural enclosure for the fiber core, thus avoiding the need for a bulky external enclosure for humidity purged atmosphere. Fibers of 5 mm and 3 mm in outer diameters with a 150 um suspended solid core and a 900 um suspended porous core respectively, were obtained by utilizing a combination of drilling and stacking techniques. Characterization of the fiber optical properties and the subwavelength imaging of the guided modes were performed using a terahertz near-field microscopy setup. Near-field imaging of the modal profiles at the fiber output confirmed the effectively single-mode behavior of such waveguides. The suspended core fibers exhibit transmission from 0.10 THz to 0.27 THz (larger core), and from 0.25 THz to 0.51 THz (smaller core). Due to the large fraction of power that is guided in the holey cladding, fiber propagation losses as low as 0.02 cm(-1) are demonstrated specifically for the small core fiber. Low-loss guidance combined with the core isolated from environmental perturbations make these all-dielectric fibers suitable for practical terahertz imaging and sensing applications. PMID- 21643168 TI - Dynamics of elliptical beams in the anomalous group-velocity dispersion regime. AB - We investigate 3D spatio-temporal focusing of elliptically-shaped beams in a bulk medium with Kerr nonlinearity and anomalous group-velocity dispersion (GVD). Strong space-time localization of the mode is observed through multi filamentation with temporal compression by a factor of 3. This behavior is in contrast to the near-zero GVD regime in which minimal pulse temporal compression is observed. Our theoretical simulations qualitatively reproduce the experimental results showing the highly localized spatio-temporal profile in the anomalous-GVD regime, which contrasts to the weakly localized pulse in the normal-GVD regime. PMID- 21643169 TI - Circular holographic video display system. AB - A circular holographic video display system reconstructs holographic video. Phase only spatial light modulators are tiled in a circular configuration in order to increase the field of view. A beam-splitter is used to align the active area of the SLMs side by side without any gap. With the help of this configuration observers can see 3D ghost-like image floating in space and can move and rotate around the object. The 3D reconstructions can be observed binocularly. Experimental results are satisfactory. PMID- 21643170 TI - On focused fields with maximum electric field components and images of electric dipoles. AB - We study focused fields which, for a given total power and a given numerical aperture, have maximum electric field amplitude in some direction in the focal point and are linearly polarized along this direction. It is shown that the optimum field is identical to the image of an electric dipole with unit magnification. In particular, the field which is the image of an electric dipole whose dipole vector is parallel to the optical axis, is identical to the field whose longitudinal component is maximum at the image point. PMID- 21643171 TI - Analytic theory of soft x-ray diffraction by Lamellar Multilayer Gratings. AB - An analytic theory describing soft x-ray diffraction by Lamellar Multilayer Gratings (LMG) has been developed. The theory is derived from a coupled waves approach for LMGs operating in the single-order regime, where an incident plane wave can only excite a single diffraction order. The results from calculations based on these very simple analytic expressions are demonstrated to be in excellent agreement with those obtained using the rigorous coupled-waves approach. The conditions for maximum reflectivity and diffraction efficiency are deduced and discussed. A brief investigation into p-polarized radiation diffraction is also performed. PMID- 21643172 TI - Rotation sensing based on a side-coupled spaced sequence of resonators. AB - A side-coupled spaced sequence of resonators (SCSSOR) displays strong dispersion with a magnitude much larger than that of conventional waveguides. We investigate the Sagnac effect in a SCSSOR structure. An explicit expression of the Sagnac phase difference of the structure is derived and discussed. The results show the sensitivity is proportional to the number and dispersion intensity of resonators. Compared with other resonator structures, one advantage is that it is not necessary to preserve the same circumference of each resonator, which is difficult to realize in practice. The results also predict a SCSSOR structure can be used for the realization of highly sensitive and compact integrated rotation sensors and gyroscopes. PMID- 21643173 TI - Terahertz digital holography using angular spectrum and dual wavelength reconstruction methods. AB - Terahertz digital off-axis holography is demonstrated using a Mach-Zehnder interferometer with a highly coherent, frequency tunable, continuous wave terahertz source emitting around 0.7 THz and a single, spatially-scanned Schottky diode detector. The reconstruction of amplitude and phase objects is performed digitally using the angular spectrum method in conjunction with Fourier space filtering to reduce noise from the twin image and DC term. Phase unwrapping is achieved using the dual wavelength method, which offers an automated approach to overcome the 2pi phase ambiguity. Potential applications for nondestructive test and evaluation of visually opaque dielectric and composite objects are discussed. PMID- 21643174 TI - Focusing a TM(01) beam with a slightly tilted parabolic mirror. AB - A parabolic mirror illuminated with an incident collimated beam whose axis of propagation does not exactly coincide with the axis of revolution of the mirror shows distortion and strong coma. To understand the behavior of such a focused beam, a detailed description of the electric field in the focal region of a parabolic mirror illuminated with a beam having a nonzero angle of incidence is required. We use the Richards-Wolf vector field equation to investigate the electric energy density distribution of a beam focused with a parabolic mirror. The explicit aberration function of this focused field is provided along with numerically calculated electric energy densities in the focal region for different angles of incidence. The location of the peak intensity, the Strehl ratio and the full-width at half-maximum as a function of the angle of incidence are given and discussed. The results confirm that the focal spot of a strongly focused beam is affected by severe coma, even for very small tilting of the mirror. This analysis provides a clearer understanding of the effect of the angle of incidence on the focusing properties of a parabolic mirror as such a focusing device is of growing interest in microscopy. PMID- 21643175 TI - Local refractive index sensitivity of plasmonic nanoparticles. AB - We report on an experimental characterization of the sensitivity of localized surface plasmons (LSP) to local changes in the refractive index at a nanometer scale. The method is based on forming a polymer mask covering different well defined areas of metallic nanoparticles and measuring the extinction peak shifts associated with the local refractive index changes. Arrays of nanoparticles (nanorod chains) are prepared using electron beam lithography and the dielectric mask is aligned with respect to the nanoparticle array in a second lithographic step. Extinction peak shifts corresponding to different positions of the mask are measured and values for the local refractive index sensitivity are deduced. A deconvolution procedure is established and used to map the local sensitivity across the surface of nanoparticle based on measured data. The experimental results are shown to correspond well with theoretical simulations obtained using the finite-difference time-domain method. The results indicate that the sensitivity is strongly correlated with the profile of the LSP electric field. PMID- 21643176 TI - Hybrid III-V semiconductor/silicon nanolaser. AB - Heterogeneous integration of III-V compound semiconductors on Silicon on Insulator is one the key technology for next-generation on-chip optical interconnects. In this context, the use of photonic crystals lasers represents a disruptive solution in terms of footprint, activation energy and ultrafast response. In this work, we propose and fabricate very compact laser sources integrated with a passive silicon waveguide circuitry. Using a subjacent Silicon On-Insulator waveguide, the emitted light from a photonic crystal based cavity laser is efficiently captured. We study experimentally the evanescent wave coupling responsible for the funneling of the emitted light into the silicon waveguide mode as a function of the hybrid structure parameters, showing that 90% of coupling efficiency is possible. PMID- 21643177 TI - Mysterious coloring: structural origin of color mixing for two breeds of Papilio butterflies. AB - The structural origin of the coloration mechanisms and related extraordinary optical properties of the wing scales of two breeds of Papilio butterflies, namely, Papilio ulysses and Papilio blumei, are explored. The precise ordered biophotonic nanostructures of the wing scales are characterized by scanning electron microscopy (SEM). Despite their structural similarities, the two breeds of Papilio butterflies do not exhibit any analogy in their optical performances. When illuminated with UV-Vis light, P. ulysses gives rise to two reflection peaks: one is from concavities, and the other is from ridges. These two spectral peaks shift their positions under different illumination angles (normal and 45 degrees incident light). In contrast, the spectra for the green scales of P. blumei give one broad reflection peak, and the peak remains the same under normal and 45 degrees incident light. The optical microscopy images indicate that the cap-shaped concavities on P. blumei's wing scales generate an abnormal bicolor reflection with a strong polarization effect. Both of these two breeds of butterflies take advantage of color mixing strategy: the blue color of P. ulysses is mixed by the colors reflected from concavities and ridges; the green color of P. blumei is produced by the biocolor reflection from concavities. The differences of their coloration mixing mechanisms and optical performances are due to the variations of their nanostructures. The investigation of the color mixing mechanisms of these biologically photonic nanostructures may offer a convenient way for fabricating optical devices based on biomimicry. PMID- 21643179 TI - Quaternary ultraviolet AlInGaN MQW laser diode performance using quaternary AlInGaN electron blocking layer. AB - The effect of polarization-matched Al(0.25)In(0.08)Ga(0.67)N electron-blocking layer (EBL) on the optical performance of ultraviolet Al(0.08)In(0.08)Ga(0.84)N/Al(0.1)In(0.01)Ga(0.84)N multi-quantum well (MQW) laser diodes (LDs) was investigated. The polarization-matched Al(0.25)In(0.08)Ga(0.67)N electron blocking layer (EBL) was employed in an attempt to reduce the polarization effect inside the active region of the diodes. The device performance which is affected by piezoelectric was studied via drift-diffusion model for carrier transport, optical gain and losses using the simulation program of Integrated System Engineering Technical Computer Aided design (ISE TCAD). The optical performance of the LD using quaternary Al(0.25)In(0.08)Ga(0.67)N EBL was compared with the LD using ternary Al(0.3)Ga(0.7)N EBL where both materials have the same energy band gap of Eg = 3.53 eV. The self-consistent ISE-TCAD simulation program results showed that the polarization-matched quaternary Al(0.25)In(0.08)Ga(0.67)N EBL is beneficial as it confines the electrons inside the quantum well region better than ternary Al(0.3)Ga(0.7)N EBL. The results indicated that the use of Al(0.25)In(0.08)Ga(0.67)N EBL has lower threshold current and higher optical intensity than those for Al(0.3)Ga(0.7)N EBL. The effect of Al(0.25)In(0.08)Ga(0.67)N EBL thickness on the performance of LDs has also been studied. Results at room temperature indicated that lower threshold current, high slope efficiency, high output power, and high differential quantum efficiency DQE occurred when the thickness of Al(0.25)In(0.08)Ga(0.67)N EBL was 0.25 um. PMID- 21643180 TI - Design of phase-shifted hybrid silicon distributed feedback lasers. AB - We present data on the design and performance analysis of phase shifted distributed feedback (DFB) lasers on the hybrid silicon platform. The lasing wavelength for various input currents and temperatures, for devices with standard quarter-wavelength, 60 MUm and 120 MUm-long phase shift are compared for mode stability and output power. The pros and cons of including a large phase shift region in the grating design are analyzed. PMID- 21643181 TI - Single mode emission and non-stochastic laser system based on disordered point sized structures: toward a tuneable random laser. AB - As an advantage, random lasers may be elaborated from a large variety of materials and do not require any cavity oscillators that usually necessitate complicated and expensive fabrication techniques. Since the feedback process of those non-conventional laser systems is provided by light interference in a disordered medium, spectral and temporal uncertainties are usually considered as an intrinsic part of their optical proprieties. We investigated random laser action under two photon absorption experiments through an auto-organized InGaN/GaN quantum-disks ensemble. Thanks to our experimental approach, we evidence random lasing based on a gain medium constituted by point-sized structures. In such context, a stabilised and individual emission mode is observed as for conventional semiconductor lasers. By controlling the emission energy of these nanostructures, a tuneable and stable random laser may be built. Moreover, our findings suggest that disordered medium should play an important role in the conception of low cost quantum dot and up conversion laser systems. PMID- 21643182 TI - Multiscale beam evolution and shaping in corrugated plasmonic systems. AB - We analyze, theoretically and experimentally, the dynamics of the wavepackets in plasmonic beaming devices, and show that the beam evolution in this class of structures is a multiscale phenomenon, which initiates in the near-field proximity of the device, develops continuously over a new length scale many times the wavelength of the light, and is completed well into the far-field of the system. We develop a quantitative description of the light evolution in the beaming structures and verify our theoretical predictions with experiments. Our analytical results are utilized to develop plasmonic geometries for shaping the mid-field beam evolution, and experimental results from these structures are demonstrated. PMID- 21643183 TI - Parallel implementation of all-digital timing recovery for high-speed and real time optical coherent receivers. AB - The digital coherent receivers combine coherent detection with digital signal processing (DSP) to compensate for transmission impairments, and therefore are a promising candidate for future high-speed optical transmission system. However, the maximum symbol rate supported by such real-time receivers is limited by the processing rate of hardware. In order to cope with this difficulty, the parallel processing algorithms is imperative. In this paper, we propose a novel parallel digital timing recovery loop (PDTRL) based on our previous work. Furthermore, for increasing the dynamic dispersion tolerance range of receivers, we embed a parallel adaptive equalizer in the PDTRL. This parallel joint scheme (PJS) can be used to complete synchronization, equalization and polarization de-multiplexing simultaneously. Finally, we demonstrate that PDTRL and PJS allow the hardware to process 112 Gbit/s POLMUX-DQPSK signal at the hundreds MHz range. PMID- 21643184 TI - Generation and long-haul transmission of polarization-switched QPSK at 42.9 Gb/s. AB - We demonstrate, for the first time, the generation and transmission of polarization-switched QPSK (PS-QPSK) signals at 42.9 Gb/s. Long-haul transmission of PS-QPSK is experimentally investigated in a recirculating loop and compared with transmission of dual-polarization QPSK (DP-QPSK) at 42.9 Gb/s per channel. A reduction in the required OSNR of 0.7 dB was found at a BER of 3.8 x 10(-3), resulting in an increase in maximum reach of more than 30% for a WDM system operating on a 50 GHz frequency grid. The maximum reach of 13640 km for WDM PS QPSK is, to the best of our knowledge, the longest distance reported for 40 Gb/s WDM transmission, over an uncompensated link, with standard fiber and amplification. PMID- 21643185 TI - Vacuum electron acceleration driven by a tightly focused radially polarized Gaussian beam. AB - Electron acceleration in vacuum driven by a tightly focused radially polarized Gaussian beam has been studied in detail. Weniger transformation method is used to eliminate the divergence of the radially polarized electromagnetic field derived from the Lax series approach. And, electron dynamics in an intense radially polarized Gaussian beam is analyzed by using the Weniger transformation field. The roles of the initial phase of the electromagnetic field and the injection angle, position and energy of electron in energy gain of electron have been studied in detail. PMID- 21643186 TI - Pulse splitting in the anomalous group-velocity-dispersion regime. AB - We investigate experimentally the role that the initial temporal profile of ultrashort laser pulses has on the self-focusing dynamics in the anomalous group velocity dispersion (GVD) regime. We observe that pulse-splitting occurs for super-Gaussian pulses, but not for Gaussian pulses. The splitting does not occur for either pulse shape when the GVD is near-zero. These observations agree with predictions based on the nonlinear Schrodinger equation, and can be understood intuitively using the method of nonlinear geometrical optics. PMID- 21643187 TI - Multispectral image enhancement for effective visualization. AB - Color enhancement of multispectral images is useful to visualize the image's spectral features. Previously, a color enhancement method, which enhances the feature of a specified spectral band without changing the average color distribution, was proposed. However, sometimes the enhanced features are indiscernible or invisible, especially when the enhanced spectrum lies outside the visible range. In this paper, we extended the conventional method for more effective visualization of the spectral features both in visible range and non visible range. In the proposed method, the user specifies both the spectral band for extracting the spectral feature and the color for visualization respectively, so that the spectral feature is enhanced with arbitrary color. The proposed color enhancement method was applied to different types of multispectral images where its effectiveness to visualize spectral features was verified. PMID- 21643188 TI - Cascadability properties of MZI-SOA-based all-optical 3R regenerators for RZ-DPSK signals. AB - We experimentally demonstrate 50 cascaded all-optical 3R regenerators over a 1,000 km transmission distance for 10-Gb/s return-to-zero differential phase shift keying (RZ-DPSK) signals. The regenerator consists of integrated Mach Zehnder interferometer (MZI) semiconductor optical amplifier (SOA) based wavelength converters. Regenerative properties and tolerance to pattern dependent effects have been studied in terms of Q-factor measurement, and error free operation with input OSNR of 20 dB/0.1 nm has also been demonstrated. PMID- 21643189 TI - Precise in-situ measurement of laser pulse intensity using strong field ionization. AB - Building on the work of Alnaser et al. [Phys. Rev. A 70, 023413 (2004)], we devise an improved method for an in-situ measurement of the peak intensity in a focused, femtosecond infrared laser pulse. The method is shown to be effective with both photoion and photoelectron imaging devices. The model used to fit the experimental data has no unphysical free parameters used in fitting. The accuracy of the fit is 4% and the overall accuracy of the measurement is 8%. PMID- 21643190 TI - Dispersion of nonlinearity and modulation instability in subwavelength semiconductor waveguides. AB - Tight confinement of light in subwavelength waveguides induces substantial dispersion of their nonlinear response. We demonstrate that this dispersion of nonlinearity can lead to the modulational instability in the regime of normal group velocity dispersion through the mechanism independent from higher order dispersions of linear waves. A simple phenomenological model describing this effect is the nonlinear Schrodinger equation with the intensity dependent group velocity dispersion. PMID- 21643191 TI - Accessing photon bunching with a photon number resolving multi-pixel detector. AB - In quantum optics and its applications, there is an urgent demand for photon number resolving detectors. Recently, there appeared multi-pixel counters (MPPC) that are able to distinguish between 1,2,..10 photons. At the same time, strong coupling between different pixels (crosstalk) hinders their photon-number resolution. In this work, we suggest a method for 'filtering out' the crosstalk effect in the measurement of intensity correlation functions. The developed approach can be expanded to the analysis of higher-order intensity correlations by using just a single MPPC. PMID- 21643192 TI - Hybrid Q-switched Yb-doped fiber laser. AB - We investigate the performance of a hybrid Q-switched (HQS) fiber laser that is constructed with a low RF-power driven acousto-optic (AO) Q-switch and an AlGaInAs semiconductor saturable absorber. Compared to a pure passively Q switched (PQS) fiber laser, the ratio of timing jitter to pulse period can be significantly reduced from 2% to 0.3% in the regime of far above threshold. On the other hand, the prelasing effect in a pure actively Q-switched fiber laser can be considerably improved. More importantly, the maximum pulse energy of the HQS fiber laser can be increased approximately 25% in comparison with the result of the PQS fiber laser. At a pump power of 24 W, the highest pulse energy is up to 0.56 mJ with the pulse duration of 50 ns at the repetition rate of 23 kHz. PMID- 21643193 TI - High-Q/V air-mode photonic crystal cavities at microwave frequencies. AB - We present results for a photonic microwave resonator designed and fabricated at 17.4 GHz with a record high Quality factor (Q=26,400) at room temperature over a mode volume smaller than one cubic wavelength. The cavity is uniquely designed to have its electric field concentrated in air, which allows for efficient coupling to free space and facilitates interactions with gaseous atomic systems and fluids. PMID- 21643194 TI - Composite, all-ceramics, high-peak power Nd:YAG/Cr(4+):YAG monolithic micro-laser with multiple-beam output for engine ignition. AB - A passively Q-switched Nd:YAG/Cr(4+):YAG micro-laser with three-beam output was realized. A single active laser source made of a composite, all-ceramics Nd:YAG/Cr(4+):YAG monolithic cavity was pumped by three independent lines. At 5 Hz repetition rate, each line delivered laser pulses with ~2.4 mJ energy and 2.8 MW peak power. The M(2) factor of a laser beam was 3.7, and stable air breakdowns were realized. The increase of pump repetition rate up to 100 Hz improved the laser pulse energy by 6% and required ~6% increase of the pump pulse energy. Pulse timing of the laser-array beams can by adjusted by less than 5% tuning of an individual line pump energy, and therefore simultaneous multi-point ignition is possible. This kind of laser can be used for multi-point ignition of an automobile engine. PMID- 21643195 TI - Light outcoupling effect in GaN light-emitting diodes via convex microstructures monolithically fabricated on sapphire substrate. AB - GaN-based light-emitting diode (LED) was fabricated on the sapphire substrate with monolithic convex microstructures (CMs) array. Using confocal scanning electroluminescence (EL), we have directly observed the strong outcoupling phenomenon of the light confined in a LED via the CMs array. This outcoupled light could be efficiently converged on the convex center through consecutive reflections at the flat area and the curved slant area of the CMs array. Compared to the conventional LED, the ray tracing simulation and far field EL results of the LED with a CM array showed efficient light extraction toward the top surface, i.e., 0-5, 40-45 and 60-65 degree by the outcoupling effect. We conclude that the outcoupled optical path via CMs is the dominant factor of the enhanced light extraction in the LED with a CM array. PMID- 21643196 TI - Polymer waveguide integrated-optic current transducers. AB - Various functional optical devices are integrated on a single chip in order to construct optical current transducers based on polarization rotated reflection interferometry, which consists of polarization maintaining 3-dB couplers, TE-pass polarizers, TE/TM polarization converters, and thermo-optic phase modulators. By virtue of the device integration, the sensor exhibited good linearity, and excellent accuracy with an error less than 0.2%. The integrated-optic device provides inherent polarization maintaining characteristics and precise controllability of the optical path length in the interferometric sensor. Single chip integration reduces the complexity of the interferometry, and enables mass production of low-cost high performance current sensors. PMID- 21643197 TI - Polarization-independent wide-angle triple-band metamaterial absorber. AB - We report the design, fabrication, and measurement of a microwave triple-band absorber. The compact single unit cell consists of three nested electric closed ring resonators and a metallic ground plane separated by a dielectric layer. Simulation and experimental results show that the absorber has three distinctive absorption peaks at frequencies 4.06 GHz, 6.73 GHz, and 9.22 GHz with the absorption rates of 0.99, 0.93, and 0.95, respectively. The absorber is valid to a wide range of incident angles for both transverse electric (TE) and transverse magnetic (TM) polarizations. The triple-band absorber is a promising candidate as absorbing elements in scientific and technical applications because of its multiband absorption, polarization insensitivity, and wide-angle response. PMID- 21643198 TI - Design and analysis of enhanced second harmonic generation in AlGaAs/AlO(x) microring waveguide. AB - In this paper, we propose and analyze a multilayer AlGaAs/AlO(x) waveguide structure for efficient microring based second harmonic generation (SHG). Quasi phase matching (QPM) and resonance conditions can be satisfied by designing the width of the waveguide. Linewidth of SHG in microring resonators is formulized and investigated and it is shown that linewidth of wavelength conversion is in the range of subnanometer. Dependence of efficiency of SHG on input power and the loss is investigated for various conditions like single and double microring structures, different coupling coefficients and radii. In low loss condition, larger radius of microring needs lower input power for efficient SHG. Our proposed double microring structure provides 100% conversion efficiency with lower input power as compared with a single microring structure. PMID- 21643199 TI - Adaptive aberration compensation for three-dimensional micro-fabrication of photonic crystals in lithium niobate. AB - We present the use of a liquid crystal spatial light modulator to correct for the refractive-index mismatch induced spherical aberration in a high refractive-index lithium niobate crystal when a low repetition rate amplified laser is used for the direct fabrication of three-dimensional micro-structures. By correcting the aberration based on experimentally determined values, we show that the size of written structures decreases dramatically, which allows the fabrication of high quality micro-structures such as three-dimensional photonic crystals. We demonstrate that, through the use of adaptive optics, the fabrication depth and the stopgap strength in the corresponding photonic crystals are increased by a factor of two to three. PMID- 21643200 TI - Polarization independence of extraordinary transmission trough 1D metallic gratings. AB - Extraordinary optical transmission of 1D metallic gratings is studied. Experimental samples are fabricated by means of Electron Beam Lithography. The optical characterization is focused on far field transmission properties and in particular on polarization dependence of the incident light. A peculiar symmetry in transmission spectra at different polarization angles is shown; this symmetry is studied both experimentally, and numerically with FEM method. A comparison between numerical and experimental data is provided. PMID- 21643201 TI - Effect of lens size on the focusing performance of plasmonic lenses and suggestions for the design. AB - We present a detailed investigation of the effect of lens size on the focusing performance of plasmonic lenses based on metallic nanoslit arrays with variable widths. The performance parameters considered include the focal length, depth of focus (DOF), full-width half-maximum (FWHM) and the maximum intensity of the focal point. 2D FDTD simulation was utilized. The results show that all the lens parameters are greatly affected by the lens size. A larger lens size, with a total phase difference of at least 2pi, will produce a better focusing behavior and a closer agreement with the design. The Fresnel number and diffraction theory can be used to explain the effect of lens size. Suggestions are provided for realization of a practical plasmonic lens using the existing nanofabrication techniques. PMID- 21643202 TI - Novel synchronous DPSK optical regenerator based on a feed-forward based carrier extraction scheme. AB - We experimentally demonstrate a novel synchronous 10.66 Gbit/s DPSK OEO regenerator which uses a feed-forward carrier extraction scheme with an injection locked laser to synchronize the regenerated signal wavelength to the incoming signal wavelength. After injection-locking, a low-cost DFB laser used at the regenerator exhibited the same linewidth characteristics as the narrow line-width transmitter laser. The phase regeneration properties of the regenerator were evaluated by emulating random Gaussian phase noise applied to the DPSK signal before the regenerator using a phase modulator driven by an arbitrary waveform generator. The overall performance was evaluated in terms of electrical eye diagrams, BER measurements, and constellation diagrams. PMID- 21643203 TI - Compensation of intra-channel nonlinear fibre impairments using simplified digital back-propagation algorithm. AB - We investigate a digital back-propagation simplification method to enable computationally-efficient digital nonlinearity compensation for a coherently detected 112 Gb/s polarization multiplexed quadrature phase shifted keying transmission over a 1,600 km link (20 x 80 km) with no inline compensation. Through numerical simulation, we report up to 80% reduction in required back propagation steps to perform nonlinear compensation, in comparison to the standard back-propagation algorithm. This method takes into account the correlation between adjacent symbols at a given instant using a weighted-average approach, and optimization of the position of nonlinear compensator stage to enable practical digital back-propagation. PMID- 21643204 TI - Ultrafast all-optical switching based on cross modulation utilizing intersubband transitions in InGaAs/AlAs/AlAsSb coupled quantum wells with DFB grating waveguides. AB - A distributed feedback Bragg grating waveguide all-optical switch design relying on the ultrafast cross modulation effect of the intersubband transitions in InGaAs/AlAs/AlAsSb coupled double quantum wells is demonstrated. The pump-induced phase modulation to the signal light is converted to intensity modulation efficiently on chip with the help of the grating structures. To our best knowledge, the switching dynamic characteristics of such design are reported for the first time. With a 400 MUm long grating waveguide, 3 dB modulation depth with switching energy of 5.5 pJ and recovery time of 4.5 ps is obtained for the switch off operation. PMID- 21643205 TI - Analysis of two-dimensional photonic crystal with anisotropic gain. AB - Photonic modes in a two-dimensional square-lattice photonic crystal (PC) with anisotropic gain are analyzed for the first time. A plane-wave expansion method is improved to include the gain, which depends on not only the position but also the propagation direction of each plane wave. The anisotropic gain varies the photonic band structure, the near-field distributions, and the gain dispersion curves through variation in PC symmetry. Low-threshold operation of a PC laser with anisotropic-gain material such as nonpolar InGaN requires that the direction of higher gain in the material aligns along the GammaX direction of the PC. PMID- 21643206 TI - Local electrical characterization of laser-recorded phase-change marks on amorphous Ge2Sb2Te5 thin films. AB - Amorphous thin films of Ge(2)Sb(2)Te(5), sputter-deposited on a thin-film gold electrode, are investigated for the purpose of understanding the local electrical conductivity of recorded marks under the influence of focused laser beam. Being amorphous, the as-deposited chalcogenide films have negligible electrical conductivity. With the aid of a focused laser beam, however, we have written on these films micron-sized crystalline marks, ablated holes surrounded by crystalline rings, and other multi-ring structures containing both amorphous and crystalline zones. Within these structures, nano-scale regions of superior local conductivity have been mapped and probed using our high-resolution, high sensitivity conductive-tip atomic force microscope (C-AFM). Scanning electron microscopy and energy-dispersive spectrometry have also been used to clarify the origins of high conductivity in and around the recorded marks. When the Ge(2)Sb(2)Te(5) layer is sufficiently thin, and when laser crystallization/ablation is used to define long isolated crystalline stripes on the samples, we find the C-AFM-based method of extracting information from the recorded marks to be superior to other forms of microscopy for this particular class of materials. Given the tremendous potential of chalcogenides as the leading media candidates for high-density memories, local electrical characterization of marks recorded on as-deposited amorphous Ge(2)Sb(2)Te(5) films provides useful information for furthering research and development efforts in this important area of modern technology. PMID- 21643207 TI - Stripe-like quasi-nondiffracting optical lattices. AB - We introduce stripe-like quasi-nondiffracting lattices that can be generated via spatial spectrum engineering. The complexity of the spatial shapes of such lattices and the distance of their almost diffractionless propagation depend on the width of their ring-like spatial spectrum. Stripe-like lattices are extended in one direction and are localized in the orthogonal one, thereby creating either straight or curved in any desired fashion optically-induced channels that may be used for optical trapping, optical manipulation, or optical lattices for quantum and nonlinear optics applications. As an illustrative example, here we show their potential for spatial soliton control. Complex networks consisting of several intersecting or joining stripe-like lattices suited to a particular application may also be constructed. PMID- 21643208 TI - Optimal design of SPP-based metallic nanoaperture optical elements by using Yang Gu algorithm. AB - An optimization method for design of SPP-based metallic nanoaperture optical elements is presented. The design process is separated into two steps: Firstly, derive the amplitude and phase modulation of isolating single slit with different width; Secondly, realize the optimal design of element by using an iteration procedure. The Yang-Gu algorithm is expanded to perform this design. Three kinds of lenses which can achieve various functions have been designed by using this method. The rigorous electromagnetical theory is employed to justify and appraise the performances of the designed elements. It has been found that the designed elements can achieve the preset functions well. This method may provide a convenient avenue to optimally design metallic diffractive optical elements with subwavelength scale. PMID- 21643209 TI - Hybrid microspheres for nonlinear Kerr switching devices. AB - Electronic Kerr effect in a polyfluorene derivative is used to reversibly switch near infrared probe beam resonantly coupled to a hybrid polymer-silica microspherical resonator. NIR pumping at 780 nm in pulsed laser regime is used for non-linear switching of the WGM resonances that shift as much as 2 GHz for 50 mW of average pump power, compared to a shift of 250 MHz for the same average pump power at CW regime. The absence of temporal drift and the magnitude of this shift confirm the Kerr nature of the switching, ruling out thermooptical effects. PMID- 21643210 TI - Phase-shifting interferometry corrupted by white and non-white additive noise. AB - The standard tool to estimate the phase of a sequence of phase-shifted interferograms is the Phase Shifting Algorithm (PSA). The performance of PSAs to a sequence of interferograms corrupted by non-white additive noise has not been reported before. In this paper we use the Frequency Transfer Function (FTF) of a PSA to generalize previous white additive noise analysis to non-white additive noisy interferograms. That is, we find the ensemble average and the variance of the estimated phase in a general PSA when interferograms corrupted by non-white additive noise are available. Moreover, for the special case of additive white noise, and using the Parseval's theorem, we show (for the first time in the PSA literature) a useful relationship of the PSA's noise robustness; in terms of its FTF spectrum, and in terms of its coefficients. In other words, we find the PSA's estimated phase variance, in the spectral space as well as in the PSA's coefficients space. PMID- 21643211 TI - In situ control and monitoring of photonic device intermixing during laser irradiation. AB - Apparatus and method for the in situ control of photonic device intermixing processes are described. The setup utilises an optical fiber splitter which delivers photons to selectively anneal the photonic device and simultaneously measures the emission spectra from the device to monitor the intermixing process in real time. The in situ monitoring of a laser annealing process for the modification of a semiconductor laser diode facet is demonstrated using the instrumentation. A progressive blueshift in the emission wavelength of the device can clearly be observed in real time while high energy photons are delivered to anneal the device facet, hence enabling the control on the degree of intermixing required. This instrumentation is also ideal for broadening of emission spectra in quantum dot and quantum well based light emitting devices such as superluminescent diodes and broadband laser. PMID- 21643212 TI - A 50 nm spatial resolution EUV imaging-resolution dependence on object thickness and illumination bandwidth. AB - In this paper we report a desk-top microscopy reaching 50 nm spatial resolution in very compact setup using a gas-puff laser plasma EUV source. The thickness of an object and the bandwidth of illuminating radiation were studied in order to estimate their quantitative influence on the EUV microscope spatial resolution. EUV images of various thickness objects obtained by illumination with variable bandwidth EUV radiation were compared in terms of knife-edge spatial resolution to study the bandwidth/object thickness parasitic influence on spatial resolution of the EUV microscope. PMID- 21643213 TI - Transverse mode tailoring in a quasi-imaging high-finesse femtosecond enhancement cavity. AB - We demonstrate a high-finesse femtosecond enhancement cavity with an on-axis obstacle. By inserting a wire with a width of 5% of the fundamental mode diameter, the finesse of F = 3400 is only slightly reduced to F = 3000. The low loss is due to the degeneracy of transverse modes, which allows for exciting a circulating field distribution avoiding the obstacle. We call this condition quasi-imaging. The concept could be used for output coupling of intracavity generated higher-order harmonics through an on-axis opening in one of the cavity mirrors. PMID- 21643214 TI - Highly efficient plasmonic interconnector based on the asymmetric junction between metal-dielectric-metal and dielectric slab waveguides. AB - A novel interconnection structure that is capable of transferring surface plasmon energy between two distanced metal-dielectric-metal waveguides is proposed. A numerical analysis indicates that the power transferring efficiency of the proposed structure approaches 88.5% for one millimeter of interconnection length. The main mechanism of the proposed interconnector is governed by Fabry-Perot resonance and a multi-mode interference effect in the junction layers. A physical analysis based on the mechanism for designing an optimized structure is also provided, which could be used to modify the proposed structure. PMID- 21643215 TI - Hybrid CATV/16-QAM OFDM in-building networks over SMF and GI-POF transport. AB - A hybrid CATV/16-QAM OFDM in-building network over a combination of single-mode fiber (SMF) and graded-index plastic optical fiber (GI-POF) transport is proposed and experimentally demonstrated with good qualities of service. In this system, a 1556 nm optical signal is directly transmitted along with two fiber spans (20-km SMF + 25-m GI-POF). Without using any wavelength conversion or bridge circuit between SMF and POF connection, error free transmissions with sufficient low bit error rate (BER) values are achieved for 2.5 Gbps/2.5 GHz and 5 Gbps/2.5 GHz OFDM signals; as well as good performances of carrier-to-noise ratio (CNR), composite second-order (CSO), and composite triple beat (CTB) are obtained for CATV one. This proposed network reveals an outstanding one with economy and convenience to be installed. PMID- 21643216 TI - Measuring the feedback parameter of a semiconductor laser with external optical feedback. AB - Feedback parameter (the C factor) is an important parameter for a semiconductor laser operating in the regime of external optical feedback. Self-mixing interferometry (SMI) has been proposed for the measurement of the parameter, based on the time-domain analysis of the output power waveforms (called SMI signals) in presence of feedback. However, the existing approaches only work for a limited range of C, below about 3.5. This paper presents a new method to measure C based on analysis of the phase signal of SMI signals in the frequency domain. The proposed method covers a large range of C values, up to about 10. Simulations and experimental results are presented for verification of the proposed method. PMID- 21643217 TI - Bistable switching in the lossy side-coupled plasmonic waveguide-cavity structures. AB - We show numerically that the lossy side-coupled plasmonic resonators can be used as bistable switches without compensation. While the internal loss imposes on the bistable characteristics by reducing the transmission contrast and raising the input power requirement, it makes the switching more available by enlarging the width of the hysteresis loop. We also correct the nonlinear transmission formula of the resonators to adapt the lossy condition. Both the theoretical and simulation results are in good agreement. PMID- 21643218 TI - Time-resolved EUV photoelectron spectroscopy of dissociating I2 by laser harmonics at 80 nm. AB - Generation of single-order laser harmonics in extreme ultraviolet (EUV) and the application to the time-resolved photoelectron spectroscopy of I(2) are demonstrated. The EUV pulses at 80 nm were generated from Kr as the 5th order harmonics of intense 400 nm laser pulses and then separated from other harmonic orders by a thin indium foil. The pump-probe photoelectron spectroscopy of I(2) in the B (3)Pi(0(u)(+)) and B" (1)Pi(1(u)) states excited by visible laser pulses at 490 nm showed a rapid increase in the yield of atomic iodine (~400 fs), reflecting the dissociation dynamics evolving simultaneously in the two excited states. PMID- 21643219 TI - Optical properties of a fabricated self-assembled bottom-up bulk metamaterial. AB - We investigate the optical properties of a true three-dimensional metamaterial that was fabricated using a self-assembly bottom-up technology. The metamaterial consists of closely packed spherical clusters being formed by a large number of non-touching gold nanoparticles. After presenting experimental results, we apply a generalized Mie theory to analyze its spectral response revealing that it is dominated by a magnetic dipole contribution. By using an effective medium theory we show that the fabricated metamaterial exhibits a dispersive effective permeability, i.e. artificial magnetism. Although this metamaterial is not yet left-handed it might serve as a starting point for achieving bulk metamaterials by using bottom-up approaches. PMID- 21643220 TI - Tunable Fabry-Perot filter using hollow-core photonic bandgap fiber and micro fiber for a narrow-linewidth laser. AB - A novel tunable fiber Fabry-Perot (FP) filter is proposed and demonstrated by using a hollow-core photonic bandgap fiber (HC-PBF) and a micro-fiber. The interference cavity is a hollow core of HC-PBF. One of the reflection mirrors is the splicing point between a section of HC-PBF and a single mode fiber. The other reflection mirror is a gold-coated end of micro-fiber that uses chemical etching process to obtain the similar diameter as the core of HC-PBF. Hence the movable mirror can be adjusted with long distance inside the hollow core of HC-PBF. Tunable FP filter is used as a mode selecting component in the reflection mode to implement stable single longitudinal mode (SLM) operation in a ring laser. With FP cavity length of 0.25 +/- 0.14 mm, the wavelength of SLM laser can be tuned over 1554-1562 nm with a tuning step of 0.2-0.3 nm, a side-mode suppression ratio (SMSR) of 32-36 dB and a linewidth of 3.0-5.1 kHz. With FP cavity length of 2.37 +/- 0.37 mm, the SLM laser can be tuned over 1557.3-1560.2 nm with a tuning step of 0.06-0.1 nm, a SMSR of 44-51 dB and a linewidth of 1.8-3.0 kHz. PMID- 21643221 TI - The tempo-spatially modulated polarization atmosphere Michelson interferometer. AB - A space-based tempo-spatially modulated polarization atmosphere Michelson interferometer (TSMPAMI) is described. It uses the relative movement between the TSMPAMI and the measured target to change optical path difference. The acquisition method of interferogram is presented. The atmospheric temperatures and horizontal winds can be derived from the optical observations. The measurement errors of the winds and temperatures are discussed through simulations. In the presence of small-scale structures of the atmospheric fields, the errors are found to be significantly influenced by the mismatch of the scenes observed by the adjacent CCD sub-areas aligned along the orbiter's track during successive measurements due to the orbital velocity and the exposure time. For most realistic conditions of the orbit and atmosphere, however, the instrument is proven suitable for measuring the atmospheric parameters. PMID- 21643222 TI - Observation of the in-plane spin separation of light. AB - We report on the observation of the spin separation of light in the plane of incidence when a linearly polarized beam is reflected or refracted at a planar dielectric interface. Remarkably, the in-plane spin separation reaches hundreds of nanometers, comparable with the transverse spin separation induced by the well known spin Hall effect of light. The observation is properly explained by considering the in-plane spread of wave-vectors. This study thus offers new insights on the spinoptics and may provide a potential method to control light in optical nanodevices. PMID- 21643223 TI - Generation of energetic femtosecond green pulses based on an OPCPA-SFG scheme. AB - Femtosecond green pulses were generated from broadband pulses centered at 800 nm and quasi-monochromatic pulses centered at 532 nm using noncollinear optical parametric chirped pulse amplification (NOPCPA) followed by sum frequency mixing. In addition to amplifying the 800-nm pulses, the NOPCPA stage pumped by a Q switched, injection seeded Nd:YAG laser also provided broadband idler pulses at 1590 nm. The signal and idler pulses were sum frequency mixed using achromatic and chirp assisted phase matching yielding pulses near 530 nm with a bandwidth of 12 nm and an energy in excess of 200 MUJ. The generated pulses were recompressed with a grating compressor to a duration of 150 fs. The technique is scalable to high energies, broader bandwidths, and shorter pulse durations with compensation for higher order chirps and dedicated engineering of the interacting beams. PMID- 21643224 TI - Partially coherent nano-focused x-ray radiation characterized by Talbot interferometry. AB - We have studied the spatial coherence properties of a nano-focused x-ray beam by grating (Talbot) interferometry in projection geometry. The beam is focused by a fixed curvature mirror system optimized for high flux density under conditions of partial coherence. The spatial coherence of the divergent exit wave emitted from the mirror focus is measured by Talbot interferometry The results are compared to numerical calculations of coherence propagation. In view of imaging applications, the magnified in-line image of a test pattern formed under conditions of partial coherence is analyzed quantitatively. Finally, additional coherence filtering by use of x-ray waveguides is demonstrated. By insertion of x-ray waveguides, the beam diameter can be reduced from typical values of 200 nm to values below 15 nm. In proportion to the reduction in the focal spot size, the numerical aperture (NA) of the projection imaging system is increased, as well as the coherence length, as quantified by grating interferometry. PMID- 21643225 TI - All-optically controllable dye-doped liquid crystal infiltrated photonic crystal fiber. AB - A novel demonstration of an all-optically controllable dye-doped liquid crystal infiltrated photonic crystal fiber (DDLCIPCF) is presented. Overall spectral transmittance of the DDLCIPCF can decrease and then increase with a concomitant red-shift of the spectrum curve with increasing irradiation time of one UV beam. Continuing irradiation of one green beam following UV illumination on the DDLCIPCF can cause the transmission spectrum to recover completely. The reversible all-optical controllability of the photonic band structure of the fiber is attributable to the isothermal planar nematic (PN)->scattering (S) >isotropic (I) and I->S->PN state transitions of the LCs via the UV-beam-induced trans->cis and green-beam-induced cis->trans back isomerizations of the azo-dye, respectively, in the cladding of the DDLCIPCF. The photoinduced appearance of the S state and the variation of the index modulation between the core and the cladding of the fiber result in the variation of overall spectral transmittance and the shift of transmission spectrum, respectively. PMID- 21643226 TI - Simultaneous guidance of slow photons and slow acoustic phonons in silicon phoxonic crystal slabs. AB - We demonstrate theoretically that photons and acoustic phonons can be simultaneously guided and slowed down in specially designed nanostructures. Phoxonic crystal waveguides presenting simultaneous phononic and photonic band gaps were designed in perforated silicon membranes that can be conveniently obtained using silicon-on-insulator technology. Geometrical parameters for simultaneous photonic and phononic band gaps were first chosen for optical wavelengths around 1550 nm, based on the finite element analysis of a perfect phoxonic crystal of circular holes. A plain core waveguide was then defined, and simultaneous slow light and elastic guided modes were identified for some waveguide width. Joint guidance of light and elastic waves is predicted with group velocities as low as c/25 and 180 m/s, respectively. PMID- 21643227 TI - Influence of self-phase modulation effect on waveform degradation and spectral broadening in optical BPSK-SSB fiber transmission. AB - Self-phase modulation (SPM) effect is analyzed in a dispersion-compensated transmission using optical BPSK single sideband (SSB) modulation. The effect was evaluated numerically using both waveform degradation and spectral degradation, clarifying that waveform degradation is induced dominantly by peak power of the quadrature component of a Hilbert-transformed signal. Eye-opening degradation of BPSK-SSB is induced by lower fiber input power than the conventional double sideband (DSB) case because the SSB-homodyne system is sensitive to phase error resulting from SPM. Spectral degradation from SPM has two phases with increasing fiber input power. In the first phase, the sideband in the suppressive frequency region expands with increasing optical power. In the second phase, the spectral envelope in the non-suppressive frequency region becomes broad, and its shape is somewhat varied. PMID- 21643228 TI - Radiation force of highly focused Lorentz-Gauss beams on a Rayleigh particle. AB - The radiation force of highly focused Lorentz-Gauss beams (LG beam) on a dielectric sphere in the Rayleigh scattering regime is theoretically studied. The numerical results show that the Lorentz-Gauss beam can be used to trap particles with the refractive index larger than that of the ambient. The radiation force distribution has been studied under different beam widths of the Lorentz part. The trapping stability under different conditions is also analyzed. PMID- 21643229 TI - Classical and quantum properties of cylindrically polarized states of light. AB - We investigate theoretical properties of beams of light with non-uniform polarization patterns. Specifically, we determine all possible configurations of cylindrically polarized modes (CPMs) of the electromagnetic field, calculate their total angular momentum and highlight the subtleties of their structure. Furthermore, a hybrid spatio-polarization description for such modes is introduced and developed. In particular, two independent Poincare spheres have been introduced to represent simultaneously the polarization and spatial degree of freedom of CPMs. Possible mode-to-mode transformations accomplishable with the help of Bconventional polarization and spatial phase retarders are shown within this representation. Moreover, the importance of these CPMs in the quantum optics domain due to their classical features is highlighted. PMID- 21643230 TI - Fast saturable absorption and 10 GHz wavelength conversion in Al-quaternary multiple quantum wells. AB - We measured the absorption recovery times in reverse biased AlInGaAs multiple quantum well material designed to emit at around 1.5 MUm wavelength. Absorption recovery times as low as 2.5 ps were found at -4V bias, with values below 5 ps consistently found for biases above 3 V. The short absorption recovery times obtained under reverse bias were confirmed by using cross-absorption modulation in the material to demonstrate wavelength conversion of a 10 GHz pulse train, showing both up and down conversion of the incident pulses. PMID- 21643231 TI - Fast-switching bistable cholesteric intensity modulator. AB - A fast-switching bistable optical intensity modulator is demonstrated. Using a dual-frequency cholesteric liquid crystal, the direct switching is achieved from the scattering focal conic state to the transparent long-pitch planar state. In comparison with the bistable cholesteric devices proposed previously, our device, characterized by its capability of direct two-way transitions between the two bistable states, possesses a very short transition time from the focal conic state to the planar state as short as 10 ms. No voltage has to be applied to sustain the optical states, making the device low energy consuming. Potential applications of this device are addressed. PMID- 21643232 TI - Self-similar pulses in coherent linear amplifiers. AB - We discover and analytically describe self-similar pulses existing in homogeneously broadened amplifying linear media in a vicinity of an optical resonance. We demonstrate numerically that the discovered pulses serve as universal self-similar asymptotics of any near-resonant short pulses with sharp leading edges, propagating in coherent linear amplifiers. We show that broadening of any low-intensity seed pulse in the amplifier has a diffusive nature: Asymptotically the pulse width growth is governed by the simple diffusion law. We also compare the energy gain factors of short and long self-similar pulses supported by such media. PMID- 21643233 TI - Multiple responses of TPP-assisted near-perfect absorption in metal/Fibonacci quasiperiodic photonic crystal. AB - Absorption properties in one-dimensional quasiperiodic photonic crystal composed of a thin metallic layer and dielectric Fibonacci multilayers are investigated. It is found that a large number of photonic stopbands can occur at the dielectric Fibonacci multilayers. Tamm plasmon polaritons (TPPs) with the frequencies locating at each photonic stopband are excited at the interface between the metallic layer and the dielectric layer, leading to almost perfect absorption for the energy of incident wave. By adjusting the length of dielectric layer with higher refractive-index or the Fibonacci order, the number of absorption peaks can be tuned effectively and enlarged significantly. PMID- 21643234 TI - Two-dimensional displacement measurement by quasi-common-optical-path heterodyne grating interferometer. AB - A method based on a specific quasi-common-optical-path (QCOP) configuration for two-dimensional displacement measurement is presented. The measurement system consists of a heterodyne light source, two-dimensional holographic grating, specially designed set of half wave plates, and lock-in amplifiers. Two measurement configurations, for single and differential detection, are designed. The sensitivity, resolution and nonlinear phase error of the differential detection type are better than those of the single detection type. The experimental results demonstrate that the QCOP interferometer has the ability to measure two-dimensional displacement while maintaining high system stability. PMID- 21643235 TI - Atmospheric correction in presence of sun glint: application to MERIS. AB - The sun glint is a major issue for the observation of ocean color from space. For sensors without a tilting capacity, the observations at sub-tropical latitudes are contaminated by the bright pattern of the specular reflexion of the sun by the wavy sea surface. Common atmospheric correction algorithms are not designed to work in these observation conditions, reducing the spatial coverage at such latitudes by nearly a half. We describe an original atmospheric correction algorithm, named POLYMER, designed to recover ocean color parameters in the whole sun glint pattern. It has been applied to MERIS data, and validated against in situ data from SIMBADA. The increase of useful coverage of MERIS measurements for ocean color is major, and the accuracy of the retrieved parameters is not significantly reduced in the presence of high sunglint, while, outside the sunglint area, it remains about the same as by using the standard algorithm. PMID- 21643236 TI - Gas jet structure influence on high harmonic generation. AB - Gas jets used as sources for high harmonic generation (HHG) have a complex three dimensional density and velocity profile. This paper describes how the profile influences the generation of extreme-UV light. As the position of the laser focus is varied along the jet flow axis, we show that the intensity of the output radiation varies by approximately three times, with the highest flux being observed when the laser is focused into the Mach disc. The work demonstrated here will aid in the optimization of HHG flux from gas jet sources. The flux increase is attributed to a density increase within the structure of the jet, which is confirmed by simultaneous imaging of atom and ion fluorescence from the jet. PMID- 21643237 TI - Light emission rate enhancement from InP MQW by plasmon nano-antenna arrays. AB - Arrays of gold single-strip and double-strip nano-antennas, with resonance in the wavelength range of 1200-1600 nm, were fabricated on the top of InGaAs/InP multi quantum well structure. Photo-luminescence from the quantum-wells was measured and shown to be enhanced by a factor of up to 9, with maximum enhancement wavelength corresponding to the nano-antennas resonance. Emission enhancement is attributed to the coupling of emitting charge-carriers to the plasmonic nano antennas, causing an estimated increase in the radiative recombination rate by a factor of ~25, thus making it dominant over non-radiative recombination. This effect will enable fast modulation of InP-based nano-emitters spontaneously emitting at telecom-wavelength. PMID- 21643238 TI - Waveguide mode filters fabricated using laser-induced forward transfer. AB - Titanium (Ti)-in-diffused lithium niobate waveguide mode filters fabricated using laser-induced forward transfer followed by thermal diffusion are presented. The mode control was achieved by adjusting the separation between adjacent Ti segments thus varying the average value of the refractive index along the length of the in-diffused channel waveguides. The fabrication details, loss measurements and near-field optical characterization of the mode filters are presented. Modeling results regarding the device performance are also discussed. PMID- 21643239 TI - Modeling of light scattering in different regimes of surface roughness. AB - The light scattering of rough metallic surfaces with roughness levels ranging from a few to several hundred nanometers is modeled and compared to experimental data. Different modeling approaches such as the classical Rayleigh-Rice vector perturbation theory and the new Generalized Harvey-Shack theory are used and critically assessed with respect to ranges of validity, accuracy, and practicability. Based on theoretical calculations and comparisons with Rigorous Coupled Wave Analysis for sinusoidal phase gratings, it is demonstrated that the approximate scatter models yield surprisingly accurate results and at the same time provide insight into light scattering phenomena. For stochastically rough metal surfaces, the predicted angles resolved scattering is compared to experimental results at 325 nm, 532 nm, and 1064 nm. In addition, the possibilities of retrieving roughness information from measured scattering data for different roughness regimes are discussed. PMID- 21643240 TI - Long-range channel plasmon polaritons in thin metal film V-grooves. AB - We numerically investigate the propagation characteristics of guided modes in a thin metal film V-groove embedded in a dielectric medium, with a particular emphasis on long-ranging channel plasmon polaritons (LR-CPPs). The LR-CPP shows several orders of magnitude larger propagation length than the previously studied short-range channel plasmon polariton (SR-CPP). Moreover, the LR-CPP possesses a peculiar mode cutoff mechanism when surrounding dielectric media are asymmetric and this makes its propagation characteristics very sensitive to index change of the surrounding dielectric media. PMID- 21643241 TI - Systematic approach to complex periodic vortex and helix lattices. AB - We present a general comprehensive framework for the description of symmetries of complex light fields, facilitating the construction of sophisticated periodic structures carrying phase dislocations. In particular, we demonstrate the derivation of all three fundamental two-dimensional vortex lattices based on vortices of triangular, quadratic, and hexagonal shape, respectively. We show that these patterns represent the foundation of complex three-dimensional lattices with outstanding helical intensity distributions which suggest valuable applications in holographic lithography. This systematic approach is substantiated by a comparative study of corresponding numerically calculated and experimentally realized complex intensity and phase distributions. PMID- 21643242 TI - Characterization of highly stable mid-IR, GaSb-based laser diodes. AB - Highly stable room-temperature, mid-IR, GaSb-based laser diodes have been characterized at various temperatures and driver currents. Up to 54 mW of output power was demonstrated in a 3150- to 3180-nm-wavelength range with <20-nm FWHM spectral width. PMID- 21643243 TI - Performance analyses of polarization demultiplexing based on constant-modulus algorithm in digital coherent optical receivers. AB - In the digital coherent optical receiver, we can achieve polarization demultiplexing in the digital domain, using a two-by-two matrix controlled by the constant-modulus algorithm (CMA). In this paper, after elucidating the physics behind CMA for polarization demultiplexing, we discuss the performance limit of CMA-based polarization demultiplexing through computer simulations. The method of improving its performance is also demonstrated. PMID- 21643244 TI - Experimental comparison of modulation formats in IM/DD links. AB - We present an experimental comparison of modulation formats for optical intensity modulated links with direct detection. Specifically, we compare OOK, QPSK on an electrical subcarrier and a new modulation format named OOPSK. The OOPSK modulation format is shown to have better sensitivity than the other modulation formats, in agreement with theoretical predictions. The impact of propagation in multimode fiber is also studied and the results show that all modulation formats have similar sensitivity penalties, with respect to the fibre length. PMID- 21643245 TI - Multichannel routing of diffraction-inhibited beams in two-dimensional photonic crystals. AB - We show that two rows of photonic lattices on each side of a narrow diffraction inhibited beam are sufficient for confinement, enabling the launching of multi beams with an interval of one lattice constant for independent propagation. A few integrated photonic circuit building blocks including arbitrary angle bends, power splitter and intersection are designed to realize flexible controls of the diffraction-inhibited beams. In addition, under wide beam illumination, incident beam power is well separated by the lattices rows, facilitating the simultaneous excitation of multiple diffraction-inhibited beams. These novel effects and building blocks offer exceptional opportunities for multichannel photonic routing. PMID- 21643246 TI - Cut-off analysis of 19-cell Yb-doped double-cladding rod-type photonic crystal fibers. AB - Yb-doped double-cladding large mode area rod-type photonic crystal fibers are a key component for power scaling in fiber laser systems. Recently, designs with 19 cell core defect, that is with 19 missing air-holes in the center of the photonic crystal cladding, have been proposed, with reported core diameter up to 100 MUm. In this paper an analysis of the cut-off wavelength of the first high-order mode in such low-NA fibers is reported, accounting for different approaches for the definition of the cladding effective index. Results have shown that taking into account the finite fiber cross-section and considering the first cladding mode of the actual fiber is mandatory to obtain a correct estimate of the cut-off wavelength. PMID- 21643247 TI - Position clamping in a holographic counterpropagating optical trap. AB - Optical traps consisting of two counterpropagating, divergent beams of light allow relatively high forces to be exerted along the optical axis by turning off one beam, however the axial stiffness of the trap is generally low due to the lower numerical apertures typically used. Using a high speed spatial light modulator and CMOS camera, we demonstrate 3D servocontrol of a trapped particle, increasing the stiffness from 0.004 to 1.5 MUN m(-1). This is achieved in the "macro-tweezers" geometry [Thalhammer, J. Opt. 13, 044024 (2011); Pitzek, Opt. Express 17, 19414 (2009)], which has a much larger field of view and working distance than single-beam tweezers due to its lower numerical aperture requirements. Using a 10*, 0.2 NA objective, active feedback produces a trap with similar effective stiffness to a conventional single-beam gradient trap, of order 1 MUN m(-1) in 3D. Our control loop has a round-trip latency of 10 ms, leading to a resonance at 20 Hz. This is sufficient bandwidth to reduce the position fluctuations of a 10 MUm bead due to Brownian motion by two orders of magnitude. This approach can be trivially extended to multiple particles, and we show three simultaneously position-clamped beads. PMID- 21643248 TI - Design rules for p-i-n diode carriers sweeping in nano-rib waveguides on SOI. AB - In this paper we present a detailed analysis of the carrier lifetime for a p-i-n junction on silicon nano-rib waveguides. Several factors determining efficiency of carriers removal from the waveguiding region will be discussed. We compare different structure geometries and spacings between p and n doped regions to show the way to optimize electrons and holes sweeping for CW nonlinear optical devices. PMID- 21643249 TI - Manufacturability estimates for optical aspheres. AB - Some simple measures of the difficulty of a variety of steps in asphere fabrication are defined by reference to fundamental geometric considerations. It is shown that effective approximations can then be exploited when an asphere's shape is characterized by using a particular orthogonal basis. The efficiency of the results allows them to be used not only as quick manufacturability estimates at the production end, but more importantly as part of an efficient design process that can boost the resulting optical systems' cost-effectiveness. PMID- 21643250 TI - Stereogram implemented with a holographic image splitter. AB - A special holographic optical element (HOE) which is used as an image splitter is developed to generate a stereogram on a 2.2-in. liquid crystal display panel. The special designed holographic optical element can be attached on the conventional panel directly to replace the traditional image splitter in a stereoscopic display panel. Experimental results show that two images corresponding to slightly different viewing angles displayed on a panel can be separated effectively and can be delivered to the right eye and left eye of an observer individually. The diffraction efficiency for individual right and left image in this developed holographic optical element is about 43%, and the contrast ratio of the diffracted images induced by cross talk is larger than 60%. Theoretical analyses show the proposed technique generates good contrast ratio and brightness performance for stereogram application. PMID- 21643251 TI - Large energy laser pulses with high repetition rate by graphene Q-switched solid state laser. AB - We demonstrated that the graphene could be used as an effective saturable absorber for Q-switched solid-state lasers. A graphene saturable absorber mirror was fabricated with large and high-quality graphene sheets deprived from the liquid phase exfoliation. Using this mirror, 105-ns pulses and 2.3-W average output power are obtained from a passively Q-switched Nd:GdVO(4) laser. The maximum pulse energy is 3.2 MUJ. The slope efficiency is as high as 37% approximating to 40% of the continue-wave laser, indicating a low intrinsic loss of the graphene. PMID- 21643252 TI - Observation of large contrast electromagnetically induced absorption resonance due to population transfer in a three-level Lambda-system interacting with three separate electromagnetic fields. AB - We describe a new scheme to induce large contrast (nearly 50%) absorption resonances using three co-propagating fields which interact with a three-level Lambda-system (obtained by the D(2) transition of (87)Rb atoms) in an N configuration scheme. A single mode laser which couples the upper ground state to the excited state of (87)Rb is phase modulated at half the hyperfine splitting frequency. The resultant three line spectrum interacts with the atomic vapor yielding a population transfer which increases the absorption by an amount which depends on the carrier to modulation side band intensity ratio. PMID- 21643253 TI - Integrated silicon-based nanoplasmonic sensor. AB - The concept of an integrated nanoplasmonic sensor implemented on a silicon substrate is presented. Developed experimental setup based on rotation of linearly polarized light provides intensity detection between two orthogonal polarizations of a He-Ne laser beam. This optical configuration yields to a sensitivity improvement and noise reduction, resulting in a resolution of 4x10( 5) Refractive Index Units. Proposed methodology is promising for the application in portable nanoplasmonic multisensing and imaging. PMID- 21643254 TI - High speed terahertz modulation from metamaterials with embedded high electron mobility transistors. AB - We present a computational and experimental study of a novel terahertz (THz) device resulting from hybridization of metamaterials with pseudomorphic high electron mobility transistors (HEMTs), fabricated in a commercial gallium arsenide (GaAs) process. Monolithic integration of transistors into each unit cell permits modulation at the metamaterial resonant frequency of 0.46 THz. Characterization is performed using a THz time-domain spectrometer (THz-TDS) and we demonstrate switching values over 30%, and THz modulation at frequencies up to 10 megahertz (MHz). Our results demonstrate the viability of incorporating metamaterials into mature semiconductor technologies and establish a new path toward achieving electrically tunable THz devices. PMID- 21643255 TI - Compact and high-resolution plasmonic wavelength demultiplexers based on Fano interference. AB - Using strong couplings of different Fabry-Perot (FP) resonators in metal insulator-metal waveguides, a compact plasmonic wavelength demultiplexer is numerically demonstrated with high wavelength resolution. In the demultiplexer, it is found that new right-angle resonators emerge with bandwidth narrower than that of the isolated FP resonators. These narrowband right-angle resonators interfere with the broadband FP resonators, resulting in Fano-line shapes in the transmission spectra. Consequently, these sharp and asymmetric Fano-line shapes considerably increase the resolution of wavelength demultiplexing, which is significantly narrower than the full width of the isolated FP resonator. PMID- 21643256 TI - Spatial coherence control of xuv supercontinuum generation by two-color laser field. AB - We investigate the spatial characteristics of xuv supercontinuum generation in the two-color laser field consisting of a fundamental and a weak second harmonic field. By optimizing the synthesized two-color field, the spatial profile of the xuv supercontinuum varies from annular-like to Gaussian-like and then the spatial quality is improved effectively, which is beneficial for its potential applications. Moreover, our calculation shows that the spatial quality of the supercontinuum is stable when the intensity of the controlling field varies in the acceptable fluctuation. PMID- 21643257 TI - Broadly tunable femtosecond mode-locking in a Tm:KYW laser near 2 MUm. AB - Efficient mode-locking in a Tm:KY(WO(4))(2) laser is demonstrated by using InGaAsSb quantum-well SESAMs. Self-starting ultrashort pulse generation was realized in the 1979-2074 nm spectral region. Maximum average output power up to 411 mW was produced around 1986 nm with the corresponding pulse duration and repetition rate of 549 fs and 105 MHz respectively. Optimised pulse durations of 386 fs were produced with an average power of 235 mW at 2029 nm. PMID- 21643258 TI - Fabrication and characterization of three-dimensional biomimetic chiral composites. AB - Here we show the fabrication and characterization of a novel class of biomimetic photonic chiral composites inspired by a recent finding in butterfly wing-scales. These three-dimensional networks have cubic symmetry, are fully interconnected, have robust mechanical strength and possess chirality which can be controlled through the composition of multiple chiral networks, providing an excellent platform for developing novel chiral materials. Using direct laser writing we have fabricated different types of chiral composites that can be engineered to form novel photonic devices. We experimentally show strong circular dichroism and compare with numerical simulations to illustrate the high quality of these three dimensional photonic structures. PMID- 21643259 TI - Strongly confined, low-threshold laser modes in organic semiconductor microgoblets. AB - We investigate lasing from high-Q, polymeric goblet-type microcavities covered by an organic semiconductor gain layer. We analyze the optical modes in the high-Q cavities using finite element simulations and present a numerical method to determine the cutoff thickness of the gain layer above which the whispering gallery modes are strongly confined in this layer. Fabricated devices show reduced lasing thresholds for increasing gain layer thicknesses, which can be explained by a higher filling factor of the optical modes in the gain layer. Furthermore, reduced lasing threshold is accompanied by a red-shift of the laser emission. PMID- 21643260 TI - Broadband near UV to visible optical activity measurement using self-heterodyned method. AB - We demonstrate that broadband electronic optical activity can be measured with supercontinuum light pulse generated by a femtosecond pump (800 nm). It is the self-heterodyned detection technique that enables us to selectively measure the real (optical rotatory dispersion, ORD) or imaginary (circular dichroism, CD) part of the chiroptical susceptibility by controlling the incident polarization state. The single-shot-based measurement that is capable of correcting power fluctuations of the continuum light is realized by using a fast CCD detector and a polarizing beam splitter. Particularly, non-differential scheme used does not rely on any polarization-switching components. We anticipate that this broadband CD/ORD spectrometry with intrinsically ultrafast time-resolution will be applied to a variety of ultrafast chiroptical dynamics studies. PMID- 21643261 TI - Extraordinarily high spectral sensitivity in refractive index sensors using multiple optical modes. AB - The extraordinary spectral sensitivity of surface plasmon resonance (SPR) sensors is commonly attributed to the modal overlap or unique dispersion of surface plasmons. In contrast to this belief, we show that such high sensitivity is due to the multi-mode nature of the sensing scheme. This concept of multi-mode sensing can be applied to dielectric systems as well in order to achieve similar extraordinary spectral sensitivity. We also show that there is a fundamental constraint between the spectral sensitivity and quality factor in such multi-mode sensing approach. PMID- 21643262 TI - Numerical investigation of mid-infrared supercontinuum generation up to 5 MUm in single mode fluoride fiber. AB - We numerically investigate mid-infrared supercontinuum generation in single mode fluoride fiber pumped by 1.56 MUm picosecond fiber lasers. To get high energy conversion efficiency in mid-infrared region, the ratio of power generated in 2.5 ~5 MUm range to the total input power for supercontinuum generation is optimized by varying the pulse width, peak power and fiber length. The long wavelength edge of the supercontinuum spectrum can be extended to 4.8 MUm in a 100 cm long fluoride fiber pumped by a 1.56 MUm fiber laser with a pulse width of 4 ps and a peak power of 100 kW, and the corresponding ratio of power generated in 2.5 ~5 MUm range to the total input power is about 44.6%. The spectral broadening is mainly caused by self-phase modulation, stimulated Raman scattering and four-wave mixing. The simulated results show that high average power supercontinuum light source in 2.5 ~5 MUm range could be obtained in fluoride fibers pumped by 1.56 MUm picosecond fiber lasers. PMID- 21643263 TI - Vertical optical antennas integrated with spiral ring gratings for large local electric field enhancement and directional radiation. AB - We propose a device for reproducible achievement of enormous enhancement of local electric field intensities. In each device, a metallic spiral ring grating is employed for efficient excitation of local surface plasmon resonance in the tiny gap of a vertically oriented optical antenna. Radiation from the optical antenna is collimated by the ring grating which facilitates efficient collection. As a numerical example, for a gold nanosphere placed one nanometer above the center of a gold spiral ring grating, our simulations predict an increase in local electric field intensity of up to seven orders of magnitude compared to planewave illumination, and collection efficiencies of up to 68% by an objective with a numerical aperture of 0.7. Single molecule SERS application is discussed. PMID- 21643264 TI - Intense dynamic bullets in a periodic lattice. AB - Femtosecond filamentation inside a periodic lattice in air is numerically shown to form intense dynamic bullets. The long propagation distance of the bullet structure is primarily attributed to the effect of the lattice that regulates the competition between linear and nonlinear spatiotemporal effects in the region of normal dispersion. PMID- 21643265 TI - Simultaneous illumination method: application in a two-channel emission fluorometer with multi-wavelength excitation. AB - We developed a new illumination method called the simultaneous illumination method. This method does not require synchronization between light sources and sensor signals, which drastically simplifies the instrumentation. As a proof-of concept, we applied this method to an oceanographic fluorometer. In principle, using this method, one can easily increase the number of characterized emission wavelengths by mounting optical sensors for as many emission wavelengths as needed. Our fluorometer has two emission-wavelength channels and twelve excitation wavelengths. The aim of this prototype is to demonstrate a viable in situ N-channel emission fluorometer with multiple wavelengths of excitation, which has not been previously realized. PMID- 21643266 TI - Investigation of the wave behaviors inside a step-modulated subwavelength metal slit. AB - In this paper, we applied the modal expansion method (MEM) to investigate the wave behaviors inside a step-modulated subwavelength metal slit. The physical mechanism of the surface plasmon polariton (SPP) transmission is investigated in detail for slit structures with either dielectric or geometric modulation. The applicability of the effective index method is discussed. Moreover, as a special case of the geometric modulation, the evanescent-wave assisted transmission is demonstrated in a thin-modulated slit. We emphasize that a complete set is necessary in order to expand the wave functions in these kinds of structures. All the calculated results by the MEM are well retrieved by the finite-difference time-domain calculation. PMID- 21643267 TI - Slow-light-enhanced codirectional couplers with negative index materials. AB - Optical codirectional coupling structures consisting of two parallel planar waveguides with negative index materials (NIMs) are systematically studied in different configurations using coupled-mode theory under the weak-coupling condition. As a result, we find that the coupling strength between copropagating optical modes can be enhanced in such structures. More importantly, both our analytical derivations and numerical simulations clearly indicate that the slow light effect in the waveguides with NIMs plays an essential role in such enhancement. The configuration with two conventional positive-index-material cores embedded in NIM claddings (or vice versa) can lead to the strongest enhancement because it can give rise to the slowest light in our scheme. Therefore, as well as offering a fundamental understanding of the slow-light effect in codirectional coupling structures with NIMs for constructing compact photonic devices, our investigations suggest a useful guideline for optimizing the design of codirectional couplers using slow-light systems for both the classical and quantum information processing and communication networks. PMID- 21643268 TI - Mechanical Kerr nonlinearities due to bipolar optical forces between deformable silicon waveguides. AB - We use an analytical method based on the perturbation of effective index at fixed frequency to calculate optical forces between silicon waveguides. We use the method to investigate the mechanical Kerr effect in a coupled-waveguide system with bipolar forces. We find that a positive mechanical Kerr coefficient results from either an attractive or repulsive force. An enhanced mechanical Kerr coefficient several orders of magnitude larger than the intrinsic Kerr coefficient is obtained in waveguides for which the optical mode approaches the air light line, given appropriate design of the waveguide dimensions. PMID- 21643269 TI - High-speed, sub-Nyquist interferometry. AB - The velocity measurement limit in dynamic interferometry is vNyq, the velocity at which the interferogram is sampled at the Nyquist limit. We show that vNyq can be exceeded by assuming continuity of the surface motion and unwrapping the velocity modulo 2vNyq. The technique was demonstrated in a high-speed speckle pattern interferometer with spatial phase stepping. Surface velocities of 4vNyq were measured experimentally. With a reduced exposure, high-speed sub-Nyquist interferometry could be implemented up to a maximum acceleration of vNyq/ts, where ts is the detector frame period. PMID- 21643270 TI - All-optical fiber anemometer based on laser heated fiber Bragg gratings. AB - A fiber-optic anemometer based on fiber Bragg gratings (FBGs) is presented. A short section of cobalt-doped fiber was utilized to make a fiber-based "hot wire" for wind speed measurement. Fiber Bragg gratings (FBGs) were fabricated in the cobalt-doped fiber using 193 nm laser pulses to serve as localized temperature sensors. A miniature all-optical fiber anemometer is constructed by using two FBGs to determine the dynamic thermal equilibrium between the laser heating and air flow cooling through monitoring the FBGs' central wavelengths. It was demonstrated that the sensitivity of the sensor can be adjusted through the power of pump laser or the coating on the FBG. Experimental results reveal that the proposed FBG-based anemometer exhibits very good performance for wind speed measurement. The resolution of the FBG-based anemometer is about 0.012 m/s for wind speed range between 2.0 m/s and 8.0 m/s. PMID- 21643271 TI - ZnGa2O4:Cr3+: a new red long-lasting phosphor with high brightness. AB - ZnGa2O4:Cr3+ is shown to be a new bright red UV excited long-lasting phosphor potentially suitable for in vivo imaging due to its 650 nm-750 nm emission range. Photoluminescence and X-ray excited radioluminescence show the 2E -> 4A2 emission lines of both ideal Cr3+ and Cr3+ distorted by a neighboring antisite defect while long-lasting phosphorescence (LLP) and thermally stimulated luminescence (TSL) almost exclusively occur via distorted Cr3+. The most intense LLP is obtained with a nominal Zn deficiency and is related to a TSL peak at 335K. A mechanism for LLP and TSL is proposed, whereby the antisite defect responsible for the distortion at Cr3+ acts as a deep trap. PMID- 21643272 TI - Terahertz scattering by subwavelength cylindrical arrays. AB - We demonstrate the use of a full-wave electromagnetic field simulator to verify terahertz (THz) transmission-mode spectroscopic measurements of periodic arrays containing subwavelength cylindrical scatterers. Many existing THz scattering studies utilize analytical solutions, which were developed for a single scatterer. For multiple scatterers, a scaling factor equal to the number of scatterers is applied, accounting for interference between far-field radiative contributions from those scatterers but not their near-field mutual coupling. Consequently, analytical solutions do not accurately verify measurements. Conversely, results from the full-wave electromagnetic field simulator elucidate our measurements well, and provide an important insight into how the scattering behavior of cylindrical scatterers is influenced by test conditions. PMID- 21643273 TI - Elastic properties of soft tissue-mimicking phantoms assessed by combined use of laser ultrasonics and low coherence interferometry. AB - Advances in the field of laser ultrasonics have opened up new possibilities in medical applications. This paper evaluates this technique as a method that would allow for rapid characterization of the elastic properties of soft biological tissue. In doing so, we propose a novel approach that utilizes a low coherence interferometer to detect the laser-induced surface acoustic waves (SAW) from the tissue-mimicking phantoms. A Nd:YAG focused laser line-source is applied to one- and two-layer tissue-mimicking agar-agar phantoms, and the generated SAW signals are detected by a time domain low coherence interferometry system. SAW phase velocity dispersion curves are calculated, from which the elasticity of the specimens is evaluated. We show that the experimental results agree well with those of the theoretical expectations. This study is the first report that a laser-generated SAW phase velocity dispersion technique is applied to soft materials. This technique may open a way for laser ultrasonics to detect the mechanical properties of soft tissues, such as skin. PMID- 21643274 TI - Actively coupled cavity ringdown spectroscopy with low-power broadband sources. AB - We demonstrate a coupling scheme for cavity enhanced absorption spectroscopy that makes use of an intracavity acousto-optical modulator to actively switch light into (and out of) a resonator. This allows cavity ringdown spectroscopy (CRDS) to be implemented with broadband nonlaser light sources with spectral power densities of less than 30MUW/nm. Although the acousto-optical element reduces the ultimate detection limit by introducing additional losses, it permits absorptivities to be measured with a high dynamic range, especially in lossy environments. Absorption measurements for the forbidden transition of gaseous oxygen in air at ~760nm are presented using a low-coherence cw-superluminescent diode. The same setup was electronically configured to cover absorption losses from 1.8*10-8cm-1 to 7.5% per roundtrip. This could be of interest in process analytical applications. PMID- 21643275 TI - Fast switching of long-pitch cholesteric liquid crystal device. AB - We propose a long-pitch cholesteric liquid crystal (ChLC) device capable of operation in both the dynamic mode and the memory mode. Fast switching between the homeotropic state and the focal conic state allows the display of moving pictures at a low operating voltage. In addition, we can write text messages on the proposed ChLC device by applying an external pressure locally to switch it from the focal conic state to the planar state. PMID- 21643276 TI - Mode instability in high power fiber amplifiers. AB - For powers exceeding a sharp threshold in the vicinity of several hundred watts the beam quality from some narrow bandwidth fiber amplifiers is severely degraded. We show that this can be caused by transverse thermal gradients induced by the amplification process. PMID- 21643277 TI - Highly flexible all-optical metamaterial absorption switching assisted by Kerr nonlinear effect. AB - A three-dimensional metamaterial nanostructure for realizing all-optical absorption switching is proposed and investigated. The structure consists of dual metallic layers for allowing near-perfect absorption due to electric and magnetic resonances, and a nonlinear Kerr-dielectric layer for actively manipulating the nanostructure absorption. The finite-difference time-domain simulation results demonstrate that, by adjusting the incident optical intensity, the metamaterial absorption can be flexibly tuned from near unity to zero. The all-optical absorption switching structure can find potential applications in actively integrated photonic circuits for thermal sensing, photo detecting, and optical imaging. PMID- 21643278 TI - Uni-traveling-carrier variable confinement waveguide photodiodes. AB - Uni-traveling-carrier waveguide photodiodes (PDs) with a variable optical confinement mode size transformer are demonstrated. The optical mode is large at the input for minimal front-end saturation and the mode transforms as the light propagates so that the absorption profile is optimized for both high-power and high-speed performance. Two differently designed PDs are presented. PD A demonstrates a 3-dB bandwidth of 12.6 GHz, and saturation currents of 40 mA at 1 GHz and 34 mA at 10 GHz. PD B demonstrates a 3-dB bandwidth of 2.5 GHz, a saturation current greater than 100 mA at 1 GHz, a peak RF output power of + 19 dBm, and a third-order output intercept point of 29.1 dBm at a photocurrent of 60 mA. PMID- 21643279 TI - Engineering photonic nanojets. AB - Photonic Nanojets are highly localized wave fields emerging directly behind dielectric microspheres; if suitably illuminated. In this contribution we reveal how different illumination conditions can be used to engineer the photonic Nanojets by measuring them in amplitude and phase with a high resolution interference microscope. We investigate how the wavelength, the amplitude distribution of the illumination, its polarization, or a break in symmetry of the axial-symmetric structure and the illumination affect the position, the localization and the shape of the photonic Nanojets. Various fascinating properties are systematically revealed and their implications for possible applications are discussed. PMID- 21643280 TI - Percussion drilling of metals using bursts of nanosecond pulses. AB - The effect of ns bursting on percussion drilling of metal is investigated experimentally and analytically, and compared with the efficiency and quality of drilling using single ns pulses. Key advantages are demonstrated, correlating well with the results from a thermal theoretical model. The 1064 nm bursts contain up to 14 pulses of various pulse widths and spacing, and at frequencies of tens of MHz within the burst. The individual pulses have pulse widths of 10 to 200 ns, and up to 12 kW peak power. Burst repetition frequency is single shot to 500 kHz. PMID- 21643281 TI - Ultrabroadband efficient intracavity XUV output coupler. AB - We report an efficient intracavity XUV output coupler based on an anti-reflection coated grazing incidence plate (GIP). Conceptually, GIP is an extension of a Brewster plate, affording low loss of the circulating fundamental light and serving as a highly efficient, extremely broadband output coupler for XUV. Due to the grazing incidence geometry, the short wavelength reflectivity can be extended to the keV range. The first GIP realized shows parameters close to the design. We discuss both the limitations of the GIP in comparison with other XUV output couplers and the applicability of the GIP extension at longer wavelengths, down to the MIR. PMID- 21643282 TI - A blind robust watermarking scheme with non-cascade iterative encrypted kinoform. AB - A blind robust watermarking scheme is proposed. A watermark is firstly transformed into a non-cascade iterative encrypted kinoform with non-cascade phase retrieve algorithm and random fractional Fourier transform (RFrFT). An iterative algorithm and Human Visual System (HVS) are both presented to adaptively embed the kinoform watermark into corresponding 2-level DWT coefficients of the cover image. The kinoform accounts for much less data amount to be embedded than regular computer-generated hologram (CGH). And the kinoform can be extracted with the only right phase key and right fractional order, and reconstructed to represent original watermark without original cover image. The experiments have shown the scheme's high security, good imperceptibility, and robustness to resist attacks such as noise, compression, filtering, cropping. PMID- 21643283 TI - Birefringence measurement of the retinal nerve fiber layer by swept source polarization sensitive optical coherence tomography. AB - A Swept Source Polarization-Sensitive Optical Coherence Tomography (SS-PS-OCT) instrument has been designed, constructed, and verified to provide high sensitivity depth-resolved birefringence and phase retardation measurements of the retinal nerve fiber layer. The swept-source laser had a center wavelength of 1059 nm, a full-width-half-max spectral bandwidth of 58 nm and an A-line scan rate of 34 KHz. Power incident on the cornea was 440 uW and measured axial resolution was 17 um in air. A multiple polarization state nonlinear fitting algorithm was used to measure retinal birefringence with low uncertainty. Maps of RNFL phase retardation in a subject measured with SS-PS-OCT compare well with those generated using a commercial scanning laser polarimetry instrument. Peak-to valley variation of RNFL birefringence given here is less than values previously reported at 840nm. PMID- 21643284 TI - THz triangulation and stand-off measurement of the refractive index. AB - We have constructed a pulsed THz imaging system based on the triangulation method. The system is capable of stand-off measurements, especially of retrieving the refractive index in a non-tactile manner even if the thickness of the object is unknown. The distance between emitter and imaged object for the presented measurements was 1.3m. We have measured a variety of samples in order to determine the capabilities and to optimize the optical properties of the instrument. PMID- 21643285 TI - Field-test of a robust, portable, frequency-stable laser. AB - We operate a frequency-stable laser in a non-laboratory environment where the test platform is a passenger vehicle. We measure the acceleration experienced by the laser and actively correct for it to achieve a system acceleration sensitivity of Deltaf / f = 11(2) * 10(-12)/g, 6(2) * 10(-12)/g, and 4(1) * 10( 12)/g for accelerations in three orthogonal directions at 1 Hz. The acceleration spectrum and laser performance are evaluated with the vehicle both stationary and moving. The laser linewidth in the stationary vehicle with engine idling is 1.7(1) Hz. PMID- 21643286 TI - Numerical study of color holographic projection using space-division method. AB - We propose a color holographic projection using the space-division method, which can reconstruct a two-dimensional color image by one hologram and avoid the superimposing of unwanted images on a wanted image. We calculated three holograms corresponding to red, green and blue, and then generated one hologram to add the three holograms. The three holograms were optimized by the Gerchberg-Saxton algorithm for improvement of reconstructed color images. We numerically evaluated the image quality of color reconstructed images in terms of the color space of YCbCr, and compared the quality of color reconstructed images by the space division method with that of reconstructed color images using another color holographic projection method. PMID- 21643287 TI - Generation of optical vortex array with transformation of standing-wave Laguerre Gaussian mode. AB - We develop a novel method of creating optical vortex array by the conversion of a standing-wave Laguerre-Gaussian (LG) mode. Theoretically, by employing the transformational relation, the standing-wave LG mode is verified to be transformed from a pair of crisscrossed Hermite-Gaussian (HG) modes, embedded with optical vortex array, consists of a TEMn,m mode and a TEMm,n mode. Due to close correspondence between the transformational relation and the mode conversion of astigmatic lenses, we successfully generate the optical vortex array by transforming a standing-wave LG mode into the crisscrossed HG modes via a pi/2 cylindrical lens mode converter. The investigation may provide useful insight in the study of the vortex light beam and its further applications. PMID- 21643288 TI - Phase-coherent detection of an optical dipole force by Doppler velocimetry. AB - We report phase-coherent Doppler detection of optical dipole forces using large ion crystals in a Penning trap. The technique is based on laser Doppler velocimetry using a cycling transition in 9Be+ near 313 nm and the center-of-mass (COM) ion motional mode. The optical dipole force is tuned to excite the COM mode, and measurements of photon arrival times synchronized with the excitation potential show oscillations with a period commensurate with the COM motional frequency. Experimental results compare well with a quantitative model for a driven harmonic oscillator. This technique permits characterization of motional modes in ion crystals; the measurement of both frequency and phase information relative to the driving force is a key enabling capability--comparable to lockin detection - providing access to a parameter that is typically not available in time-averaged measurements. This additional information facilitates discrimination of nearly degenerate motional modes. PMID- 21643289 TI - Electrically driven hybrid Si/III-V Fabry-Perot lasers based on adiabatic mode transformers. AB - We report the first demonstration of an electrically driven hybrid silicon/III-V laser based on adiabatic mode transformers. The hybrid structure is formed by two vertically superimposed waveguides separated by a 100-nm-thick SiO2 layer. The top waveguide, fabricated in an InP/InGaAsP-based heterostructure, serves to provide optical gain. The bottom Si-waveguides system, which supports all optical functions, is constituted by two tapered rib-waveguides (mode transformers), two distributed Bragg reflectors (DBRs) and a surface-grating coupler. The supermodes of this hybrid structure are controlled by an appropriate design of the tapers located at the edges of the gain region. In the middle part of the device almost all the field resides in the III-V waveguide so that the optical mode experiences maximal gain, while in regions near the III-V facets, mode transformers ensure an efficient transfer of the power flow towards Si-waveguides. The investigated device operates under quasi-continuous wave regime. The room temperature threshold current is 100 mA, the side-mode suppression ratio is as high as 20 dB, and the fiber-coupled output power is ~7 mW. PMID- 21643290 TI - Demonstration of a cylindrically symmetric second-order nonlinear fiber with self assembled organic surface layers. AB - We report the fabrication and characterization of a cylindrically symmetric fiber structure that possesses significant and thermodynamically stable second-order nonlinearity. Such fiber structure is produced through nanoscale self-assembly of nonlinear molecules on a silica fiber taper and possesses full rotational symmetry. Despite its highly symmetric configuration, we observed significant second harmonic generation (SHG) and obtained good agreement between experimental results and theoretical predictions. PMID- 21643291 TI - Space-time block code based MIMO encoding for large core step index plastic optical fiber transmission systems. AB - The performance of Space-Time Block Codes combined with Discrete MultiTone modulation applied in a Large Core Step-Index POF link is examined theoretically. A comparative study is performed considering several schemes that employ multiple transmitters/receivers and a fiber span of 100 m. The performance enhancement of the higher diversity order configurations is revealed by application of a Margin Adaptive Bit Loading technique that employs Chow's algorithm. Simulations results of the above schemes, in terms of Bit Error Rate as a function of the received Signal to Noise Ratio, are provided. An improvement of more than 6 dB for the required electrical SNR is observed for a 3 * 1 configuration, in order to achieve a 10(-3) BER value, as compared to a conventional Single Input Single output scheme. PMID- 21643292 TI - Photon coincidences in spontaneous parametric down-converted radiation excited by a blue LED in bulk LiIO3 crystal. AB - We report on experimental and numerical investigation of two-photon coincidence properties of the parametric spontaneous down-converted field excited by a high brightness blue LED in bulk lithium iodate crystal. Ratio of up to 11.5% of coincidence, which cannot be attributed to classical coincidences, to single photon counts was recorded at the outputs of multimode fibers, demonstrating well preserved biphoton property. This result, combined with practically useful power of the source, suggests its possible application for a class of quantum experiments. PMID- 21643293 TI - Phase retrieval in X-ray phase-contrast imaging suitable for tomography. AB - In-line phase-contrast X-ray imaging provides images where both absorption and refraction contribute. For quantitative analysis of these images, the phase needs to be retrieved numerically. There are many phase-retrieval methods available. Those suitable for phase-contrast tomography, i.e., non-iterative phase-retrieval methods that use only one image at each projection angle, all follow the same pattern though derived in different ways. We outline this pattern and use it to compare the methods to each other, considering only phase-retrieval performance and not the additional effects of tomographic reconstruction. We also outline derivations, approximations and assumptions, and show which methods are similar or identical and how they relate to each other. A simple scheme for choosing reconstruction method is presented, and numerical phase-retrieval performed for all methods. PMID- 21643294 TI - Single-cell isolation using a DVD optical pickup. AB - A low-cost single-cell isolation system incorporating a digital versatile disc burner (DVD RW) optical pickup has been developed. We show that these readily available modules have the required laser power and focusing optics to provide a steady Gaussian beam capable of optically trapping micron-sized colloids and red blood cells. Utility of the pickup is demonstrated through the non-destructive isolation of such particles in a laminar-flow based microfluidic device that captures and translates single microscale objects across streamlines into designated channel exits. In this, the integrated objective lens focusing coils are used to steer the optical trap across the channel, resulting in the isolation of colloids and red blood cells using a very inexpensive off-the-shelf optical component. PMID- 21643295 TI - Field test of quantum key distribution in the Tokyo QKD Network. AB - A secure communication network with quantum key distribution in a metropolitan area is reported. Six different QKD systems are integrated into a mesh-type network. GHz-clocked QKD links enable us to demonstrate the world-first secure TV conferencing over a distance of 45km. The network includes a commercial QKD product for long-term stable operation, and application interface to secure mobile phones. Detection of an eavesdropper, rerouting into a secure path, and key relay via trusted nodes are demonstrated in this network. PMID- 21643296 TI - Complex relaxation rates of the Drude metals and their effects on the lifetime and symmetry of plasmon resonances. AB - The Drude model for metal is extended to include complex relaxation rates. As a test for what happens to the surface plasmon resonances with such metals, the lifetime is examined for propagating waves across a single planar metal dielectric interface. By analytically solving the dispersion relation being fourth-order in the complex frequency, group-velocity dispersion and quality factors are explicitly found. Due to the symmetry breaking between the forward and backward waves, standing waves are not allowed in general. PMID- 21643297 TI - Phase noise suppression of optical OFDM signals in 60-GHz RoF transmission system. AB - The dispersion-induced phase noise (PN) in an OFDM RoF system at 60 GHz leads to not only subcarrier phase rotation (PRT) but also intercarrier interference (ICI) to severely degrade the transmission performance, when a commercial cost effective DFB laser with the linewidth of several MHz is adopted. To mitigate both PRT and ICI, a post PN suppression algorithm is proposed, and it does not require any bandwidth-consuming pilot tone. For a 25.78-Gbps 16-QAM OFDM RoF signal using the laser with 1.8-MHz linewidth, employing the algorithm can extend the maximum transmission distance which corresponds to 3-dBm power penalty at the BER of 2 * 10(-3) from 75 km to more than 115 km, i.e. 50% increment of transmission distance. PMID- 21643298 TI - Mechanisms for extraordinary optical transmission through bull's eye structures. AB - We analyze both experimentally and theoretically the physical mechanisms that determine the optical transmission through deep sub-wavelength bull's eye structures (concentric annular grooves surrounding a circular hole). Our analysis focus on the transmission resonance as a function of the distance between the central hole and its nearest groove. We find that, for that resonance, each groove behaves almost independently, acting as an optical cavity that couples to incident radiation, and reflecting the surface plasmons radiated by the other side of the same cavity. It is the constructive contribution at the central hole of these standing waves emitted by independent grooves which ends up enhancing transmission. Also for each groove the coupling and reflection coefficients for surface plasmons are incorporated into a phenomenological Huygens-Fresnel model that gathers the main mechanisms to enhance transmission. Additionally, it is shown that the system presents a collective resonance in the electric field that does not lead to resonant transmission, because the fields radiated by the grooves do not interfere constructively at the central hole. PMID- 21643299 TI - Dynamics of Raman soliton during supercontinuum generation near the zero dispersion wavelength of optical fibers. AB - We observe unique dynamics of Raman soliton during supercontinuum process when an input pulse experiences initially normal group-velocity dispersion with a negative dispersion slope. In this situation, the blue components of the spectrum form a Raman soliton that moves faster than the input pulse and eventually decelerates because of Raman-induced frequency downshifting. In the time domain, the soliton trajectory bends and becomes vertical when the Raman shift ceases to occur as the spectrum of Raman soliton approaches the zero dispersion point. Parts of the red components of the pulse spectrum are captured by the Raman soliton through cross-phase modulation and they travel with it. The influence of soliton order, input chirp and dispersion slope on the dynamics of Raman soliton is discussed thoroughly. PMID- 21643300 TI - Sierpinski fractal plasmonic antenna: a fractal abstraction of the plasmonic bowtie antenna. AB - A new class of bowtie antennas with Sierpinski fractal features is proposed for sensing molecular vibration modes in the near- to mid-infrared. These antennas offer a compact device footprint and an enhanced confinement factor compared to a bowtie antenna. Through extensive simulations, it is shown that these characteristics are related to the ability of this fractal geometry to become polarized. Simulation results demonstrate that these antennas may be tuned between 700 nm <= lambda <= 3.4 um and that electric field enhancement by 56 is possible at the center of the antenna gap. PMID- 21643301 TI - Integrated GaN photonic circuits on silicon (100) for second harmonic generation. AB - We demonstrate second order optical nonlinearity in a silicon architecture through heterogeneous integration of single-crystalline gallium nitride (GaN) on silicon (100) substrates. By engineering GaN microrings for dual resonance around 1560 nm and 780 nm, we achieve efficient, tunable second harmonic generation at 780 nm. The chi2 nonlinear susceptibility is measured to be as high as 16 +/- 7 pm/V. Because GaN has a wideband transparency window covering ultraviolet, visible and infrared wavelengths, our platform provides a viable route for the on chip generation of optical wavelengths in both the far infrared and near-UV through a combination of chi2 enabled sum-/difference-frequency processes. PMID- 21643302 TI - Highly sensitive and simple method for refractive index sensing of liquids in microstructured optical fibers using four-wave mixing. AB - We present both experimental measurements and simulations for a simple fiber optical liquid refractive index sensor, made using only commercially available components and without advanced postprocessing of the fiber. Despite the simplicity, we obtain the highest sensitivity experimentally demonstrated to date for aqueous solutions (refractive index around 1.33), which is relevant for extensions to biosensing. The sensor is based on measuring the spectral shift of peaks arising from four-wave mixing (FWM), when filling the holes of a microstructured fiber with different liquid samples and propagating nanosecond pulses through the silica-core of the fiber. To the best of our knowledge, this is also the first experiment where a liquid is filled into the holes of a solid core microstructured fiber to control the phase-match conditions for FWM. PMID- 21643303 TI - Fanolike resonance due to plasmon excitation in linear chains of metal bumps. AB - We report the transmission anomaly in a modified slit grating, which is dressed, on the slit sidewalls, with the linear chains of metal bumps. An asymmetric lineshape, which is characteristic of the Fano resonance, has been found in a narrow frequency range of the spectrum. The effect can be attributed to the interference between nonresonant background transmission and resonant plasmonic wave excitation in the linear chains. The dispersion of chain plasmon mode has been suggested, enabling the dynamic tuning of spectral position of the Fano effect. PMID- 21643304 TI - In situ polarized micro-Raman investigation of periodic structures realized in liquid-crystalline composite materials. AB - In situ polarized micro-Raman Spectroscopy has been utilized to determine the liquid crystal configuration inside a periodic liquid crystalline composite structure made of polymer slices alternated to films of liquid crystal. Liquid crystal, Norland Optical Adhesive (NOA-61) monomer and its polymerized form have been investigated separately. The main Raman features, used as markers for the molecular orientation estimation, have been identified. In situ polarized Raman spectra indicate that the orientation of the liquid crystal director inside the structure is perpendicular to its polymeric slices. Results show the usefulness of in situ polarized micro-Raman spectroscopy to investigate liquid crystalline composite structures. PMID- 21643305 TI - Temporal correlation of photons following frequency up-conversion. AB - We demonstrate an approach to measure temporal correlations of photons in the near infrared range using frequency up-conversion. In this approach, the near infrared signal photons are converted into the visible range, in which highly efficient silicon avalanche photodiodes are used to perform the temporal correlation measurements. A coherent light source and a pseudo-thermal light source were used in the experiment. The results are in agreement with theoretical values and those obtained from measurements directly made using superconducting nanowire single photon detectors. We conclude that the temporal correlation (up to 4th order) of photons was preserved in the frequency up-conversion process. We further theoretically and experimentally studied the influence of the dark counts on the measurement. The setup uses commercially available components and achieves high total detection efficiency (~26%). PMID- 21643306 TI - Wavelength-swept Yb-fiber master-oscillator-power-amplifier with 70 nm rapid tuning range. AB - A continuous-wave all-polarization maintaining ytterbium-doped fiber master oscillator power amplifier, with a tuning range of 70 nm addressable at tuning rates of up to 20 nm/ms, is described. Up to 10 W of linearly polarized output was generated with an amplified spontaneous emission content of less than 0.2% throughout the tuning range. PMID- 21643307 TI - Sub-micron free-standing metal slabs with dielectric nano-voids of arbitrary shapes embedded beneath atomically-flat surface. AB - Thin metal slabs with plasmonic nano-voids buried within the skin depth (< 25 nm) of surface plasmon polaritons have been of theoretical as well as technical interests for many years due to its unique optical properties such as sharp absorbance dips and anti-crossing plasmonic dispersion characteristics. Unfortunately, such interesting plasmonic properties have not been experimentally reproduced, especially in the UV-Vis regime, owing to the involuntary surface roughness occurred in systems fabricated using conventional techniques. Here, we describe a versatile cryogenic-stripping approach for encapsulating a monolayer of nano-voids of virtually any arbitrary shapes underneath an atomically-smooth (delta < 0.55 nm) surface of a free-standing metal slab. By artificially varying the topography of the capping metal surface from ultra-smooth to moderately rough, we show structural symmetricity in a nano-void-metal system can render the overall plasmonic responses becoming profoundly influenced by the surface smoothness. The current fabrication technique is thus of primary importance to the preparation of any kind of smooth nano-void-passivated metal slabs. PMID- 21643308 TI - Optical design and imaging performance testing of a 9.6-mm diameter femtosecond laser microsurgery probe. AB - We present the optical design of a 9.6-mm diameter fiber-coupled probe for combined femtosecond laser microsurgery and nonlinear optical imaging. Towards enabling clinical use, we successfully reduced the dimensions of our earlier 18 mm microsurgery probe by half, while improving optical performance. We use analytical and computational models to optimize the miniaturized lens system for off-axis scanning aberrations. The optimization reveals that the optical system can be aberration-corrected using simple aspheric relay lenses to achieve diffraction-limited imaging resolution over a large field of view. Before moving forward with custom lenses, we have constructed the 9.6-mm probe using off-the shelf spherical relay lenses and a 0.55 NA aspheric objective lens. In addition to reducing the diameter by nearly 50% and the total volume by 5 times, we also demonstrate improved lateral and axial resolutions of 1.27 um and 13.5 um, respectively, compared to 1.64 um and 16.4 um in our previous work. Using this probe, we can successfully image various tissue samples, such as rat tail tendon that required 2-3 * lower laser power than the current state-of-the-art. With further development, image-guided, femtosecond laser microsurgical probes such as this one can enable physicians to achieve the highest level of surgical precision anywhere inside the body. PMID- 21643309 TI - Efficient generation of periodic and quasi-periodic non-diffractive optical fields with phase holograms. AB - The superposition of multiple plane waves with appropriate propagation vectors generates a periodic or quasi-periodic non-diffractive optical field. We show that the Fourier spectrum of the phase modulation of this field is formed by two disjoint parts, one of which is proportional to the Fourier spectrum of the field itself. Based on this result we prove that the non-diffractive field can be generated, with remarkable high accuracy and efficiency, in a Fourier domain spatial filtering setup, using a synthetic phase hologram whose transmittance is the phase modulation of the field. In a couple of cases this result is presented analytically, and in other cases the proof is computational and experimental. PMID- 21643310 TI - Inverse design of a three-dimensional nanophotonic resonator. AB - The inverse design of a three-dimensional nanophotonic resonator is presented. The design methodology is computationally fast (10 minutes on a standard desktop workstation) and utilizes a 2.5-dimensional approximation of the full three dimensional structure. As an example, we employ the proposed method to design a resonator which exhibits a mode volume of 0.32(lambda/n)3 and a quality factor of 7063. PMID- 21643311 TI - Laser-induced thermophoresis of individual particles in a viscous liquid. AB - This paper presents a detailed investigation of the motion of individual micro particles in a moderately-viscous liquid in direct response to a local, laser induced temperature gradient. By measuring particle trajectories in 3D, and comparing them to a simulated temperature profile, it is confirmed that the thermally-induced particle motion is the direct result of thermophoresis. The elevated viscosity of the liquid provides for substantial differences in the behavior predicted by various models of thermophoresis, which in turn allows measured data to be most appropriately matched to a model proposed by Brenner. This model is then used to predict the effective force resulting from thermophoresis in an optical trap. Based on these results, we predict when thermophoresis will strongly inhibit the ability of radiation pressure to trap nano-scale particles. The model also predicts that the thermophoretic force scales linearly with the viscosity of the liquid, such that choice of liquid plays a key role in the relative strength of the thermophoretic and radiation forces. PMID- 21643312 TI - All-optical binary phase-coded UWB signal generation for multi-user UWB communications. AB - An all-optical incoherent scheme for generation of binary phase-coded ultra wideband (UWB) signals is proposed and experimentally demonstrated. The binary phase coding is performed based on all-optical phase modulation in a semiconductor optical amplifier (SOA) and phase modulation to intensity modulation (PM-IM) conversion in a fiber delay interferometer (DI) that serves as a multichannel frequency discriminator. By locating the phase-modulated light waves at the positive and negative slopes of the DI transmission spectra, binary phase encoded UWB codes (0 and pi) are generated. We also experimentally demonstrate a bipolar UWB coding system with a code length of 4, operating at 1.25 Gb/s. And the decoding is analyzed as well. Our proposed system has potential application in future high-speed UWB impulse radio over optical fiber access networks. PMID- 21643313 TI - Investigation on multi-core fibers with large Aeff and low micro bending loss. AB - To realize large effective area (Aeff) multi-core fibers (MCFs), the design to suppress the cross-talk and the influence of the cladding diameter on the micro bending loss were investigated. As a result, the MCFs with large Aeff over 100 MUm(2) and low micro bending loss were successfully fabricated. The results indicate the importance of fiber design to realize large Aeff MCFs including fiber diameters, which largely affect the micro bending loss property. Additionally, MCF with large Aeff, low attenuation loss and suppressed cross-talk was successfully realized by optimizing the fiber design. The cross-talk properties could be estimated by the simulation based on the coupling power theory taking the influences of the longitudinal fluctuation of core diameter into account. PMID- 21643314 TI - Broadband infrared meanderline reflective quarter-wave plate. AB - We present a novel reflective quarter-wave plate comprised of subwavelength meanderline elements. The device is operational over the long-wave infrared (LWIR) spectrum, with significant spectral and angular bandwidths. Power reflection is approximately 70% over the majority of the LWIR. Efficient conversion from a 45 degrees linear polarization state into circular polarization is demonstrated from finite-element electromagnetic simulations and from broadband polarimetric measurements. PMID- 21643315 TI - Bandwidth measurement of multimode fibers using system level bit error rate testing. AB - We propose and demonstrate a novel bandwidth measurement method for multimode fibers through measuring the bit error rate and power penalty associated with the testing system. The relationship between system performance and bandwidth limitation is established through the use of well characterized electric filters. With the calibration information, bandwidths of actual fibers were measured. The results were compared with those from other methods. The benefit of the BER based bandwidth measurement method is discussed. PMID- 21643316 TI - Experimental demonstration of frequency pulling in single-pass free-electron lasers. AB - Frequency pulling is a well-known phenomenon in standard laser physics, leading to a shift of the laser frequency when the cavity and maximum gain frequencies are detuned. In this letter we present the first experimental demonstration of frequency pulling in single-pass free-electron lasers. Measurements are performed using the single-pass free-electron laser installed on the Elettra storage ring. PMID- 21643317 TI - Image processing method for laser damage probability measurement by single-shot of laser pulse. AB - An experiment for high efficiency laser-induced damage probability measurement has been implemented using a periodic binary phase grating. With such a grating, a laser beam is transformed into an ensemble array of Gaussian-like spots, which is known as the Fresnel image of the grating. A scientific CCD camera is used to image the laser spot array as well as the damage of the coating sample. The image data is then processed to obtain the peak fluence distribution of the laser spot array based on the calibrated CCD grayscale. By comparing the image of the damaged coating sample with that of the laser spot array, the damage probability of the coating sample can be precisely determined by the use of single-shot of laser pulse. PMID- 21643318 TI - High-rate quantum key distribution over 100 km using ultra-low-noise, 2-GHz sinusoidally gated InGaAs/InP avalanche photodiodes. AB - We have demonstrated quantum key distribution (QKD) over 100 km using single photon detectors based on InGaAs/InP avalanche photodiodes (APDs). We implemented the differential phase shift QKD (DPS-QKD) protocol with electrically cooled and 2-GHz sinusoidally gated APDs. The single-photon detector has a dark count probability of 2.8 * 10(-8) (55 counts per second) with a detection efficiency of 6 %, which enabled us to achieve 24 kbit/s secure key rate over 100 km of optical fiber. The DPS-QKD system offers better performances in a practical way than those achieved using superconducting single-photon detectors. Moreover, the distance that secure keys against the general individual attacks can be distributed has been extended to 160 km. PMID- 21643319 TI - Spectral properties and mechanism of instability of nanoengineered silver blocks. AB - The instability of silver nanoblocks under atmospheric conditions is investigated. The localized surface plasmon resonance band of the silver nanoblocks shows a red shift, broadening, and damping with increasing storage time under atmospheric conditions. The change in spectral properties of silver nanoblocks is considered to be due to sulfidation of silver and structural breakage of silver nanoblock based on scanning electron microscope observation and numerical simulation. The effect of aspect ratio of silver nanoblocks on the change in spectral properties of the nanoengineered silver blocks is also discussed. PMID- 21643320 TI - Real-time terahertz material characterization by numerical three-dimensional optimization. AB - Terahertz time domain spectroscopy allows for characterization of dielectrics even in cases where the samples thickness is unknown. However, a parameter extraction over a broad frequency range with simultaneous thickness determination is time consuming using conventional algorithms due to the large number of optimization steps. In this paper we present a novel method to extract the data. By employing a three dimensional optimization algorithm the calculation effort is significantly reduced while preserving the same accuracy level as conventional approaches. The presented method is even fast enough to be used in imaging applications. PMID- 21643321 TI - Type-2 fuzzy thresholding using GLSC histogram of human visual nonlinearity characteristics. AB - Image thresholding is one of the most important approaches for image segmentation and it has been extensively used in many image processing or computer vision applications. In this paper, a new image thresholding method is presented using type-2 fuzzy sets based on GLSC histogram of human visual nonlinearity characteristics (HVNC).The traditional GLSC histogram takes the image spatial information into account in a different way from two-dimensional histogram. This work refines the GLSC histogram by embedding HVNC into GLSC histogram. To select threshold based on the redefined GLSC histogram, we employ the type-2 fuzzy set, whose membership function integrates the effect of pixel gray value and local spatial information to membership value. The type-2 fuzzy set is subsequently transformed into a type-1 fuzzy set for fuzziness measure computation via type reduction. Finally, the optimal threshold is obtained by minimizing the fuzziness of the type-1 fuzzy set after an exhaustive search. The experiment on different types of images demonstrates the effectiveness and the robustness of our proposed thresholding technique. PMID- 21643322 TI - Microlens array fabricated using electrohydrodynamic instability and surface properties. AB - We fabricated polarization-dependent and polarization-independent microlens arrays (MLA) through the electrohydrodynamic instability of the optically anisotropic organic layer. The anisotropic flow induced by the instability of the organic layer leads to making the lens profile on the patterned electrode. We can easily control the polarization dependence of the MLA by controlling the surface alignment properties, even with the optically anisotropic organic layer. This method is a straightforward, fast, and reliable process for MLA fabrication since it does not require cumbersome developing and molding processes. PMID- 21643323 TI - Tailored resonator coupling for modifying the terahertz metamaterial response. AB - We experimentally and numerically study the nature of coupling between laterally paired terahertz metamaterial split-ring resonators. Coupling is shown to modify the inductive-capacitive (LC) resonances resulting in either red or blue shifting. Results indicate that tuning of the electric and magnetic coupling parameters may be accomplished not by changing the orientation or density of SRRs, but by a design modification at the unit cell level. These experiments illustrate additional degrees of freedom in tuning the electromagnetic response, which offers a path to more robust metamaterial designs. PMID- 21643324 TI - Displacement Talbot lithography: a new method for high-resolution patterning of large areas. AB - Periodic micro and nano-structures can be lithographically produced using the Talbot effect. However, the limited depth-of-field of the self-images has effectively prevented its practical use, especially for high-resolution structures with periods less than 1 micrometer. In this article we show that by integrating the diffraction field transmitted by a grating mask over a distance of one Talbot period, one can obtain an effective image that is independent of the absolute distance from the mask. In this way high resolution periodic patterns can be printed without the depth-of-field limitation of Talbot self images. For one-dimensional patterns the image obtained is shown to be related to the convolution of the mask transmission function with itself. This technique, which we call Displacement Talbot Lithography (DTL), enables high-resolution photolithography without the need for complex and expensive projection optics for the production of periodic structures like diffraction gratings or photonic crystals. Experimental results showing the printing of linear gratings and an array of holes on a hexagonal lattice are presented. PMID- 21643325 TI - Design of phase-shifting algorithms by fine-tuning spectral shaping. AB - To estimate the modulating wavefront of an interferogram in Phase Shifting Interferometry (PSI) one frequently uses a Phase Shifting Algorithm (PSA). All PSAs take as input N phase-shifted interferometric measures, and give an estimation of their modulating phase. The first and best known PSA designed explicitly to reduce a systematic error source (detuning) was the 5-steps, Schwider-Hariharan (SH-PSA) PSA. Since then, dozens of PSAs have been published, designed to reduce specific data error sources on the demodulated phase. In Electrical Engineering the Frequency Transfer Function (FTF) of their linear filters is their standard design tool. Recently the FTF is also being used to design PSAs. In this paper we propose a technique for designing PSAs by fine tuning the few spectral zeroes of a PSA to approximate a template FTF spectrum. The PSA's spectral zeroes are moved (tuned) while gauging the plot changes on the resulting FTF's magnitude. PMID- 21643326 TI - Inverse scattering problem in presence of a conducting cylinder. AB - This paper deals with the inverse scattering problem, in which a conducting cylinder is placed near samples that are to be reconstructed. Due to multiple scattering effect, the radius of the conducting cylinder and its distance to samples play an important role in inverse scattering problem. The paper investigates the role of the conducting cylinder under different arrangement of transmitting/receiving antennas. Numerical simulations show that with a proper arrangement of the cylinder and transmitting/receiving antennas, it is possible to achieve high-resolution reconstruction results with fewer antennas than when the conducting cylinder is absent. PMID- 21643327 TI - Complex-coupled photonic crystal THz lasers with independent loss and refractive index modulation. AB - Compared to near infra-red photonic crystal (PhC) band-edge lasers, achieving vertical emission with quantum cascade (QC) material operating in the THz range needs dedicated engineering because the TM polarized emission of QCLs favors in plane emitting schemes and the currently used double plasmon waveguide, prevents vertical light extraction. We present an approach with independent refractive index and extraction losses modulation. The extraction losses are obtained with small extracting holes located at appropriate positions. The modal operation of the PhC is shown to critically depend on the external losses introduced. Very high surface emission power for optimum loss extractor design is achieved. PMID- 21643328 TI - Evaluation of nonlinear absorptivity in internal modification of bulk glass by ultrashort laser pulses. AB - Thermal conduction model is presented, by which nonlinear absorptivity of ultrashort laser pulses in internal modification of bulk glass is simulated. The simulated nonlinear absorptivity agrees with experimental values with maximum uncertainty of +/- 3% in a wide range of laser parameters at 10 ps pulse duration in borosilicate glass. The nonlinear absorptivity increases with increasing energy and repetition rate of the laser pulse, reaching as high as 90%. The increase in the average absorbed laser power is accompanied by the extension of the laser-absorption region toward the laser source. Transient thermal conduction model for three-dimensional heat source shows that laser energy is absorbed by avalanche ionization seeded by thermally excited free-electrons at locations apart from the focus at pulse repetition rates higher than 100 kHz. PMID- 21643329 TI - Laser erasable implanted gratings for integrated silicon photonics. AB - In this work we experimentally demonstrate laser erasable germanium implanted Bragg gratings in SOI. Bragg gratings are formed in a silicon waveguide by ion implantation induced amorphization, and are subsequently erased by a contained laser thermal treatment process. An extinction ratio up to 24dB has been demonstrated in transmission for the fabricated implanted Bragg gratings with lengths up to 1000um. Results are also presented, demonstrating that the gratings can be selectively removed by UV pulsed laser annealing, enabling a new concept of laser erasable devices for integrated photonics. PMID- 21643330 TI - Fast-gated single-photon counting technique widens dynamic range and speeds up acquisition time in time-resolved measurements. AB - In many time-domain single-photon measurements, wide dynamic range (more than 5 orders of magnitude) is required in short acquisition time (few seconds). We report on the results of a novel technique based on a time-gated Single-Photon Avalanche Diode (SPAD) able to increase the dynamic range of optical investigations. The optical signal is acquired only in well-defined time intervals. Very fast 200-ps gate-ON transition is used to avoid the undesired strong signal, which can saturate the detector, hide the fainter useful signal and reduce the dynamic range. In experimental measurements, we obtained a dynamic range approaching 8 decades in few minutes of acquisition. PMID- 21643331 TI - Multi-modality optical neural imaging using coherence control of VCSELs. AB - Neural optical imaging can evaluate cortical hemodynamic fluctuations which reflect neural activity and disease state. We evaluate the use of vertical-cavity surface-emitting lasers (VCSELs) as illumination source for simultaneous imaging of blood flow and tissue oxygenation dynamics ex vivo and in vivo and demonstrate optical imaging of blood flow changes and oxygenation changes in response to induced ischemia. Using VCSELs we show a rapid switching from a single-mode to a special multi-mode rapid current sweep operation and noise values reduced to within a factor of 40% compared to non-coherent LED illumination. These VCSELs are promising for long-term portable continuous monitoring of brain dynamics in freely moving animals. PMID- 21643332 TI - Real-time image stabilization for arbitrary motion blurred image based on opto electronic hybrid joint transform correlator. AB - An efficient approach was put forward to keep real-time image stabilization based on opto-electronic hybrid processing, by which image motion vector can be effectively detected and point spread function (PSF) was accurately modeled instantaneously, it will alleviate greatly the complexity of image restoration algorithm. The approach applies to arbitrary motion blurred images. We have also constructed an image stabilization measurement system. The experimental results show that the proposed method has advantages of real time and preferable effect. PMID- 21643333 TI - Camera calibration under optimal conditions. AB - Different methods based on photogrammetry or self-calibration exist to calibrate intrinsic and extrinsic camera parameters and also for data pre- and post processing. From a practical viewpoint, it is quite difficult to decide which calibration method gives accurate results and even whether any data processing is necessary. This paper proposes a set of optimal conditions to resolve the calibration process accurately. The calibration method uses several images of a 2D pattern. Optimal conditions define the number of points and the number of images to resolve the calibration accurately, as well as positions and orientations from where images should be taken. PMID- 21643334 TI - Fabrication of an asymmetric Bragg coupler-based polymeric filter with a single grating waveguide. AB - In this work, a first report on fabricating an asymmetric Bragg coupler-based filter on polymeric waveguides without input-waveguide grating was revealed. The fabrication process we developed was using holographic interference techniques, capillary effect, soft lithography, and micro molding process. The transmission dip of about -9.2 dB and the 3 dB transmission bandwidth of about 0.125 nm were obtained from a filter. PMID- 21643335 TI - High-sensitivity DPSK receiver for high-bandwidth free-space optical communication links. AB - A high-sensitivity modem and high-dynamic range optical automatic gain controller (OAGC) have been developed to provide maximum link margin and to overcome the dynamic nature of free-space optical links. A sensitivity of -48.9 dBm (10 photons per bit) at 10 Gbps was achieved employing a return-to-zero differential phase shift keying based modem and a commercial Reed-Solomon forward error correction system. Low-noise optical gain was provided by an OAGC with a noise figure of 4.1 dB (including system required input loses) and a dynamic range of greater than 60 dB. PMID- 21643336 TI - Low-cost rapid miniature optical pressure sensors for blast wave measurements. AB - This paper presents an optical pressure sensor based on a Fabry-Perot (FP) interferometer formed by a 45 degrees angle polished single mode fiber and an external silicon nitride diaphragm. The sensor is comprised of two V-shape grooves with different widths on a silicon chip, a silicon nitride diaphragm released on the surface of the wider V-groove, and a 45 degrees angle polished single mode fiber. The sensor is especially suitable for blast wave measurements: its compact structure ensures a high spatial resolution; its thin diaphragm based design and the optical demodulation scheme allow a fast response to the rapid changing signals experienced during blast events. The sensor shows linearity with the correlation coefficient of 0.9999 as well as a hysteresis of less than 0.3%. The shock tube test demonstrated that the sensor has a rise time of less than 2 us from 0 kPa to 140 kPa. PMID- 21643337 TI - Time resolved chirp measurements of gain switched semiconductor laser using a polarization based optical differentiator. AB - We present a novel implementation of the "phase reconstruction using optical ultra fast differentiation" (PROUD) technique and apply it to characterize the time resolved chirp of a gain switched semiconductor laser. The optical temporal differentiator is a fiber based polarization interferometer. The method provides a fast and simple recovery of the instantaneous frequency from two temporal intensity measurements, obtained by changing the spectral response of the interferometer. Pulses with different shapes and durations of hundreds of picoseconds are fully characterized in amplitude and phase. The technique is validated by comparing the measured pulse spectra with the reconstructed spectra obtained from the intensity and the recovered phase. PMID- 21643338 TI - Pulse train fluorescence technique for measuring triplet state dynamics. AB - We report on a method to study the dynamics of triplet formation based on the fluorescence signal produced by a pulse train. Basically, the pulse train acts as sequential pump-probe pulses that precisely map the excited-state dynamics in the long time scale. This allows characterizing those processes that affect the population evolution of the first excited singlet state, whose decay gives rise to the fluorescence. The technique was proven to be valuable to measure parameters of triplet formation in organic molecules. Additionally, this single beam technique has the advantages of simplicity, low noise and background-free signal detection. PMID- 21643339 TI - Q-switching and efficient harmonic generation from a single-mode LMA photonic bandgap rod fiber laser. AB - We demonstrate a Single-Mode (SM) Large-Mode-Area (LMA) ytterbium-doped PCF rod fiber laser with stable and close to diffraction limited beam quality with 110W output power. Distributed-Mode-Filtering (DMF) elements integrated in the cladding of the rod fiber provide a robust spatial mode with a Mode-Field Diameter (MFD) of 59MUm. We further demonstrate high pulse energy Second-Harmonic Generation (SHG) and Third Harmonic Generation (THG) using a simple Q-switched single-stage rod fiber laser cavity architecture reaching pulse energies up to 1mJ at 515nm and 0.5mJ at 343nm. PMID- 21643340 TI - Maskless lithography using silicon oxide etch-stop layer induced by megahertz repetition femtosecond laser pulses. AB - In this study we report a new method for maskless lithography fabrication process by a combination of direct silicon oxide etch-stop layer patterning and wet alkaline etching. A thin layer of etch-stop silicon oxide of predetermined pattern was first generated by irradiation with high repetition (MHz) ultrafast (femtosecond) laser pulses in air and at atmospheric pressure. The induced thin layer of silicon oxide is used as an etch stop during etching process in alkaline etchants such as KOH. Our proposed method has the potential to enable low-cost, flexible, high quality patterning for a wide variety of application in the field of micro- and nanotechnology, this technique can be leading to a promising solution for maskless lithography technique. A Scanning Electron Microscope (SEM), optical microscopy, Micro-Raman, Energy Dispersive X-ray (EDX) and X-ray diffraction spectroscopy were used to analyze the silicon oxide layer induced by laser pulses. PMID- 21643341 TI - Low absorption liquid crystals for mid-wave infrared applications. AB - A partially fluorinated terphenyl liquid crystal with low absorption in both mid wave infrared (MWIR) and near IR regions is developed and its properties evaluated. This compound exhibits a nematic phase (although only about 2 degrees C), reasonably high birefringence (Deltan~0.2), low visco-elastic coefficient, and modest dielectric anisotropy (Deltaepsilon = -2.7). This compound serves as an important first example for future development of low-loss MWIR liquid crystals and devices. PMID- 21643342 TI - Experimental comparison of coherent polarization-switched QPSK to polarization multiplexed QPSK for 10 * 100 km WDM transmission. AB - Polarization-switched quadrature phase-shift keying has been demonstrated experimentally at 40.5 Gb/s with a coherent receiver and digital signal processing. Compared to polarization-multiplexed QPSK at the same bit rate, its back-to-back sensitivity at 10(-3) bit-error-ratio shows 0.9dB improvement, and it tolerates about 1.6 dB higher launch power for 10 * 100 km, 50 GHz-spaced WDM transmission allowing 1 dB penalty in required optical-signal-to-noise ratio relative to back-to-back. PMID- 21643343 TI - Deep ultraviolet diamond Raman laser. AB - We present a synchronously pumped diamond Raman laser operating at 275.7 nm pumped by the 4th harmonic of a mode locked Nd:YVO4 laser. The laser had a threshold pump pulse energy of 5.8 nJ and generated up to 0.96 nJ pulses at 10.3% conversion efficiency. The results agree well with a numerical model that includes two-photon absorption of the pump and Stokes beams and uses a Raman gain coefficient of diamond of 100 cm/GW. We also report on the observation of nanometer scale two-photon assisted etching of the diamond crystal surfaces. PMID- 21643344 TI - THz emission characteristics from p/n junctions with metal lines under non-bias conditions for LSI failure analysis. AB - We have investigated the characteristics of THz emissions from p/n junctions with metallic lines under non-bias conditions. The waveforms, spectra, and polarizations depend on the length and shape of the lines. This indicates that the transient photocurrents from p/n junctions flow into the metallic lines that emit THz waves and act as an antenna. We have successfully demonstrated the non contact inspection of open defects of multi-layered interconnects in a large scale integrated circuit using the laser THz emission microscope (LTEM). The p/n junctions connected to the defective interconnects can be identified by comparing the LTEM images of normal and defective circuits. PMID- 21643345 TI - Limits of applicability of polarization sensitive reflectometry. AB - We present a detailed theoretical analysis of the measurement limits of polarization sensitive reflectometry, imposed by spatial resolution and measurement accuracy. The limits are conveniently represented in a map of constraints. We also describe and experimentally verify a procedure that allows to measure spin profiles of single-mode fibers with spin rates exceeding the measurable range of the reflectometer. The technique consists in twisting the fiber to locally unwind the spin. PMID- 21643346 TI - Optical quenching of photoconductivity in CdSe single nanowires via waveguiding excitation. AB - We demonstrate broadband optical quenching of photoconductivity in CdSe single nanowires with low excitation power. Using 1550-nm-wavelength light with 10-nW power for waveguiding excitation, we observe a typical responsivity of 0.5 A/W for quenching the photoconductivity established by 10-uW 660-nm-wavelength background light in a 403-nm-diameter CdSe nanowire, with detectable limit of the quenching power down to pW level at room temperature, which is several orders of magnitude lower than those reported previously. This large quenching effect originates from the enhanced light-defect interaction in the nanowires via waveguiding excitation. These results open new opportunities for noninvasive characterization of deep-level defect states in low-dimensional semiconductor nanomaterials, and novel optoelectronic applications of semiconductor nanowires such as high-sensitive broadband photodetection. PMID- 21643347 TI - Higher extinction ratio circular polarizers with hetero-structured double-helical metamaterials. AB - We have recently shown circular polarizers with the homo-structured double helical metamaterials, which have broader operation bands than those of the single-helical structures [Opt. Lett. 35, 2588 (2010)]. However, trying to get more operation bands deteriorates the extinction ratio. In this paper, we proposed circular polarizers with hetero-structured double-helical metamaterials. The extinction ratios of these circular polarizers are two times higher than those with homo-structured double-helical metamaterials. Furthermore, we qualitatively explained the phenomenon of the higher extinction ratio from the viewpoint of the interaction between the two helix-wires in a double-helical unit. PMID- 21643349 TI - Surface relief model for photopolymers without cover plating. AB - Relief surface changes provide interesting possibilities for storing diffractive optical elements on photopolymers and are an important source of information to characterize and understand the material behaviour. In this paper we present a 3 dimensional model based on direct measurements of parameters to predict the relief structures generated on the material. This model is successfully applied to different photopolymers with different values of monomer diffusion. The importance of monomer diffusion in depth is also discussed. PMID- 21643350 TI - Fano-type spectral asymmetry and its control for plasmonic metal-insulator-metal stub structures. AB - We use coupled mode theory (CMT) to analyze a metal-insulator-metal (MIM) plasmonic stub structure, to reveal the existence of asymmetry in its transmittance spectra. Including the effect of the near field contribution for the stub structure, the observed asymmetry is interpreted as Fano-type interference between the quasi-continuum T-junction-resonator local-modes and discrete stub eigenmodes. Based on the asymmetry factor derived from the CMT analysis, methods to control transmittance asymmetry are also demonstrated. PMID- 21643351 TI - Multistage VIPA etalons for high-extinction parallel Brillouin spectroscopy. AB - We demonstrate a high-resolution high-extinction parallel spectrometer for Brillouin spectroscopy of turbid samples. Cascading multiple VIPA etalons in the cross-axis configuration allowed us to achieve a high extinction ratio of up to 80 dB with sub-GHz resolution. Using a three-stage VIPA, we obtained the Brillouin spectra from Intralipid solutions at concentrations up to 10%. PMID- 21643352 TI - Improved optical pulse propagation in water using an evolutionary algorithm. AB - Optical pulse propagation in water is experimentally investigated using an evolutionary algorithm (EA) to control the shape of an optical pulse. The transmission efficiency (ratio of output to input optical power) is maximized by searching the combined amplitude and phase space governing an optical pulse shaper. The transmission efficiency of each tested pulse is physically determined by experiment during the course of the optimization. Combining the EA with an experiment in this manner is a powerful means of improving some figure of merit because no analytical or computational model is required-we optimize directly given the physics of the experiment. In addition, the EA is capable of efficiently searching a large parameter space. Here, we demonstrate improved linear optical pulse propagation near 800nm. Our results demonstrate a pulse with a dramatically narrower bandwidth that coincides with a local absorption minimum (near 800 nm) implying that the transmission efficiency is dominated by water's absorption spectrum. PMID- 21643353 TI - GI-core polymer parallel optical waveguide with high-loss, carbon-black-doped cladding for extra low inter-channel crosstalk. AB - Graded-index (GI) polymer parallel optical waveguides with high-absorption, carbon-black-doped cladding are fabricated using the preform method in order to reduce the inter-channel crosstalk. The waveguides exhibit a lower inter-channel crosstalk (<-69.3 dB) than optically-transparent-clad waveguides (~-33.7 dB) and maintain low propagation loss (0.029 dB/cm). We characterize the waveguides with different concentration of carbon black in order to confirm the required concentration (required absorption loss) for keeping the inter-channel crosstalk low enough. In addition, carbon-black-doped waveguides are fabricated directly on a substrate by means of a soft-lithography method. Crosstalk is sufficiently decreased despite the high scattering loss of the core material, while insertion loss is not increased. Furthermore, we fabricate a waveguide with a high scattering-loss cladding to confirm the origin of low crosstalk in carbon-black doped waveguides. We confirm that high scattering loss of cladding is not necessarily as effective for crosstalk reduction as high absorption loss of cladding. PMID- 21643354 TI - Novel concept for ultracompact polarization splitter-rotator based on silicon nanowires. AB - A novel concept for an ultracompact polarization splitter-rotator is proposed by utilizing a structure combining an adiabatic taper and an asymmetrical directional coupler. The adiabatic taper structure is singlemode at the input end while it becomes multimode at the other end. When light propagates along the adiabatic taper structure, the TM fundamental mode launched at the narrow end is efficiently (close to 100%) converted to the first higher-order TE mode at the wide end because of the mode coupling between them. By using an asymmetrical directional coupler that has two adjacent waveguides with different core widths, the first higher-order TE mode is then coupled to the TE fundamental mode of the adjacent narrow waveguide. On the other hand, the input TE polarization does not change when it goes through the adiabatic taper structure. In the region of the asymmetrical directional coupler, the TE fundamental mode in the wide waveguide is not coupled to the adjacent narrow waveguide because of phase mismatch. In this way, TE- and TM- polarized light are separated while the TM fundamental mode is also converted into the TE fundamental mode. A design example of the proposed polarization splitter-rotator is given by using silicon-on-insulator nanowires and the total length of the device is less than 100MUm. Furthermore, only a one mask process is needed for the fabrication process, which is compatible with the standard fabrication for the regular photonic integrated circuits based on SOI nanowires. PMID- 21643355 TI - Photolithographically fabricated low-loss asymmetric silicon slot waveguides. AB - We demonstrate low-loss asymmetric slot waveguides in silicon-on-insulator (SOI). 130 and 180 nm wide slots were fabricated with a 248 nm stepper, in 200 nm thick silicon. An asymmetric waveguide design is shown to expand the range in which the TE0 mode is guided and suppress the TE1 mode, while still maintaining a sharp concentration of electric field in the center of the slot. Optical propagation losses of 2 dB/cm or less are shown for asymmetric slot waveguides with 130 nm wide slots and 320 and 100 nm wide arms. PMID- 21643356 TI - Analysis of low F-number dual micro-axilens array with binary structures by rigorous electromagnetic theory. AB - We investigate a two-dimensional low F-number dual micro-axilens array with binary structures based on a rigorous electromagnetic theory. The focal characteristics of a binary dual micro-axilens array (BDMA), including axial performances (focal depth and focal shift) and transverse performances (focal spot size and diffraction efficiency), have been analyzed in detail for different F-numbers, different incident polarization (TE and TM) waves, and different distances between micro-axilens. Numerical results reveal that the interference effect of a BDMA is not very evident, which is useful for building a BDMA with a high fill factor, and the focal characteristics of a BDMA are sensitive to the polarization of an incident wave. The comparative results have also shown that the diffraction efficiency of a BDMA will increase and the focal spot size of a BDMS will decrease when the F-number increases, for both TE polarization and TM polarization, respectively. It is expected that this investigation will provide useful insight into the design of micro-optical elements with high integration. PMID- 21643357 TI - 36 GHz submicron silicon waveguide germanium photodetector. AB - We present two effective approaches to improve the responsivity of high speed waveguide-based Ge photodetectors integrated on a 0.25 MUm silicon-on-insulator (SOI) platform. The main cause of poor responsivity is identified as metal absorption from the top contact to Ge. By optimizing Ge thickness and offsetting the contact window, we have demonstrated that the responsivity can be improved from 0.6A/W to 0.95 A/W at 1550 nm with 36 GHz 3 dB bandwidth. We also demonstrate that a wider device with double offset contacts can achieve 1.05 A/W responsivity at 1550 nm and 20 GHz 3 dB bandwidth. PMID- 21643358 TI - Analysis of the carrier-suppressed single-sideband modulators used to mitigate Rayleigh backscattering in carrier-distributed PON. AB - By using the carrier-suppressed single-sideband (CS-SSB) modulation, the Rayleigh backscattering (RB) experienced by the uplink signal can be effectively mitigated due to the reduction of the spectral overlap between the uplink signal and the distributed optical carrier. In this work, we first introduce the theoretical analysis of the CS-SSB generation using the dual-drive MZM (DD-MZM)-based and a dual-parallel MZM (DP-MZM)-based optical networking units (ONUs). Due to the different modulation mechanisms of the two CS-SSB modulations, the frequency components of the generated CS-SSB signals are also different. The transmission performance and the dispersion tolerance of the uplink signals generated by the two CS-SSB modulators are also analyzed and discussed. PMID- 21643359 TI - High index contrast semiconductor ARROW and hybrid ARROW fibers. AB - We investigate the guidance properties of two photonic crystal fibers that have been fabricated by filling the holes of a silica template with hydrogenated amorphous silicon inclusions. The first is an all-solid fiber that guides light via an antiresonant reflecting optical waveguiding mechanism and the second is only partially filled so that it guides light by a hybrid of modified total internal reflection and antiresonant reflecting optical waveguiding. It will be shown that, by selectively filling the silica template to leave an unfilled internal ring of holes, the fiber's confinement loss can be reduced significantly. This novel fiber design in which the light guided in the silica core can be modified by the semiconductor cladding provides a route to integrating functional semiconductor fibers with existing silica fiber infrastructures. PMID- 21643360 TI - Texturing of titanium (Ti6Al4V) medical implant surfaces with MHz-repetition-rate femtosecond and picosecond Yb-doped fiber lasers. AB - We propose and demonstrate the use of short pulsed fiber lasers in surface texturing using MHz-repetition-rate, microjoule- and sub-microjoule-energy pulses. Texturing of titanium-based (Ti6Al4V) dental implant surfaces is achieved using femtosecond, picosecond and (for comparison) nanosecond pulses with the aim of controlling attachment of human cells onto the surface. Femtosecond and picosecond pulses yield similar results in the creation of micron-scale textures with greatly reduced or no thermal heat effects, whereas nanosecond pulses result in strong thermal effects. Various surface textures are created with excellent uniformity and repeatability on a desired portion of the surface. The effects of the surface texturing on the attachment and proliferation of cells are characterized under cell culture conditions. Our data indicate that picosecond pulsed laser modification can be utilized effectively in low-cost laser surface engineering of medical implants, where different areas on the surface can be made cell-attachment friendly or hostile through the use of different patterns. PMID- 21643361 TI - Generation of high-temperature and low-density plasmas for improved spectral resolutions in laser-induced breakdown spectroscopy. AB - Improved spectral resolutions were achieved in laser-induced breakdown spectroscopy (LIBS) through generation of high-temperature and low-density plasmas. A first pulse from a KrF excimer laser was used to produce particles by perpendicularly irradiating targets in air. A second pulse from a 532 nm Nd:YAG laser was introduced parallel to the sample surface to reablate the particles. Optical scattering from the first-pulse plasmas was imaged to elucidate particle formation in the plasmas. Narrower line widths (full width at half maximums: FWHMs) and weaker self-absorption were observed from time-integrated LIBS spectra. Estimation of plasma temperatures and densities indicates that high temperature and low density can be achieved simultaneously in plasmas to improve LIBS resolutions. PMID- 21643362 TI - Tracking integration in concentrating photovoltaics using laterally moving optics. AB - In this work the concept of tracking-integrated concentrating photovoltaics is studied and its capabilities are quantitatively analyzed. The design strategy desists from ideal concentration performance to reduce the external mechanical solar tracking effort in favor of a compact installation, possibly resulting in lower overall cost. The proposed optical design is based on an extended Simultaneous Multiple Surface (SMS) algorithm and uses two laterally moving plano convex lenses to achieve high concentration over a wide angular range of +/-24 degrees . It achieves 500* concentration, outperforming its conventional concentrating photovoltaic counterparts on a polar aligned single axis tracker. PMID- 21643363 TI - Enhanced angular characteristics of indium tin oxide nanowhisker-coated silicon solar cells. AB - Omnidirectional and broadband light harvesting is critical to photovoltaics due to the sun's movement and its wide spectral range of radiation. In this work, we demonstrate distinctive indium-tin-oxide nanowhiskers that achieve superior angular and spectral characteristics for crystalline silicon solar cells using angle-resolved reflectance spectroscopy. The solar-spectrum weighted reflectance is well below 6% for incident angles of up to 70 degrees and for the wavelength range between 400nm and 1000nm. As a result, the nanowhisker coated solar cell exhibits broadband quantum efficiency characteristics and enhanced short-circuit currents for large angles of incidence. PMID- 21643364 TI - Design of high-efficient freeform LED lens for illumination of elongated rectangular regions. AB - We propose a method for the design of an optical element generating the required irradiance distribution in a rectangular area with a large aspect ratio. Application fields include streetlights, the illumination of halls or corridors, and so forth. The design assumes that the optical element has a complex form and contains two refractive surfaces. The first one converts a spherical beam from the light source to a cylindrical beam. The second one transforms an incident cylindrical beam and generates the required irradiance distribution in the target plane. Two optical elements producing a uniform irradiance distribution from a Cree(r) XLamp(r) source in rectangular regions of 17 m * 4 m and 17 m * 2 m are designed. The light efficiency of the designed optical element is larger than 83%, whereas the irradiance nonuniformity is less than 9%. PMID- 21643365 TI - Metal-nitride-oxide-semiconductor light-emitting devices for general lighting. AB - The potential for application of silicon nitride-based light sources to general lighting is reported. The mechanism of current injection and transport in silicon nitride layers and silicon oxide tunnel layers is determined by electro-optical characterization of both bi- and tri-layers. It is shown that red luminescence is due to bipolar injection by direct tunneling, whereas Poole-Frenkel ionization is responsible for blue-green emission. The emission appears warm white to the eye, and the technology has potential for large-area lighting devices. A photometric study, including color rendering, color quality and luminous efficacy of radiation, measured under various AC excitation conditions, is given for a spectrum deemed promising for lighting. A correlated color temperature of 4800K was obtained using a 35% duty cycle of the AC excitation signal. Under these conditions, values for general color rendering index of 93 and luminous efficacy of radiation of 112 lm/W are demonstrated. This proof of concept demonstrates that mature silicon technology, which is extendable to low-cost, large-area lamps, can be used for general lighting purposes. Once the external quantum efficiency is improved to exceed 10%, this technique could be competitive with other energy-efficient solid-state lighting options. PMID- 21643366 TI - Design and global optimization of high-efficiency solar thermal systems with tungsten cermets. AB - Solar thermal, thermoelectric, and thermophotovoltaic (TPV) systems have high maximum theoretical efficiencies; experimental systems fall short because of losses by selective solar absorbers and TPV selective emitters. To improve these critical components, we study a class of materials known as cermets. While our approach is completely general, the most promising cermet candidate combines nanoparticles of silica and tungsten. We find that 4-layer silica-tungsten cermet selective solar absorbers can achieve thermal transfer efficiencies of 84.3% at 400 K, and 75.59% at 1000 K, exceeding comparable literature values. Three layer silica-tungsten cermets can also be used as selective emitters for InGaAsSb-based thermophotovoltaic systems, with projected overall system energy conversion efficiencies of 10.66% at 1000 K using realistic design parameters. The marginal benefit of adding more than 4 cermet layers is small (less than 0.26%, relative). PMID- 21643367 TI - Glancing angle deposited ITO films for efficiency enhancement of a-Si:H/MUc-Si:H tandem thin film solar cells. AB - Indium tin oxide (ITO) thin films with relatively high transparency and low absorption are prepared by glancing angle deposition (GLAD) method and their effect on the device performance of a-Si:H/MUc-Si:H tandem thin film solar cells is theoretically investigated by applying the experimentally measured physical data of the fabricated films to the simulation parameters. The GLAD of ITO produces inclined porous columnar nanostructures due to the atomic shadowing effect. With increasing the incident flux angle, the columns are increasingly inclined, thus resulting in the improved transmission property as well as the decrease of the refractive index and extinction coefficient because of enhanced porosity within the film. Furthermore, the antireflection characteristics are improved over a wide wavelength range of 300-1100 nm. For a-Si:H/MUc-Si:H tandem thin film solar cell structure incorporated with the 0 degrees ITO/80 degrees ITO bi-layer structure, the conversion efficiency (eta) of 13.6% is obtained from simulation under AM1.5g illumination, indicating an efficiency improvement compared to the device with the 0 degrees ITO/0 degrees ITO bi-layer structure (i.e. eta = 12.58%). PMID- 21643368 TI - New paradigm of multi-chip white LEDs: combination of an InGaN blue LED and full down-converted phosphor-converted LEDs. AB - This study introduces innovative multi-chip white LED systems that combine an InGaN blue LED and green/red or green/amber/red full down-converted, phosphor conversion LEDs (pc-LEDs). Efficient green, amber, and red full down-converted pc LEDs were fabricated by simply capping a long-wave pass filter (LWPF) on top of LED packing associated with each corresponding powder phosphor. The principal advantage of this type of color-mixing approach in newly developed multi-chip white LEDs based on colored pc-LEDs is thought to be dynamic control of the chromaticity and better light quality. In addition, the color-mixing approach improves the low efficacy of green/amber LEDs in the "green gap" wavelength; reduces the wide color/efficacy variations of each primary LED with at different temperatures and currents; and improves the low color rendering indexes of the traditional color-mixing approach in red, green, and blue (RGB) multi-chip white LEDs. PMID- 21643369 TI - Flat Fresnel doublets made of PMMA and PC: combining low cost production and very high concentration ratio for CPV. AB - The linear chromatic aberration (LCA) of several combinations of polycarbonates (PCs) and poly (methyl methacrylates) (PMMAs) as singlet, hybrid (refractive/diffractive) lenses and doublets operating with wavelengths between 380 and 1600 nm - corresponding to a typical zone of interest of concentrated photovoltaics (CPV) - are compared. Those comparisons show that the maximum theoretical concentration factor for singlets is limited to about 1000 * at normal incidence and that hybrid lenses and refractive doublets present a smaller LCA increasing the concentration factor up to 5000 * and 2 * 10(6) respectively. A new achromatization equation more useful than the Abbe equation is also presented. Finally we determined the ideal position of the focal point as a function of the LCA and the geometric concentration which maximizes the flux on the solar cell. PMID- 21643370 TI - Efficiency enhancement of flexible organic light-emitting devices by using antireflection nanopillars. AB - We present an antireflection structure consisted of irregular nanopillars to increase light extraction efficiency of flexible organic light-emitting devices. The nanopillars were made by imprinting the anodized aluminum oxide on polycarbonate substrates. The thermal viscosity effect formed the nanopillars with tapered shapes. Such nanopillars show excellent antireflection properties for a wide range of incident angles and wavelengths. The normal transmittance was improved from 85.5% to 95.9% for 150-nm-height nanopillars. The transmittance was greatly improved from 52.8% to 89.1% at 60 degrees incident angle. With this antireflection structure, the device efficiency was improved 69% as compared to devices with flat substrates. Due to wide-angle antireflection, the image contrast ratio was also significantly improved. PMID- 21643371 TI - Controlling Fano lineshapes in plasmon-mediated light coupling into a substrate. AB - Metal nanoparticles are efficient resonant plasmonic scatterers for light, and, if placed on top of a high-index substrate, can efficiently couple light into the substrate. This coupling, however, strongly depends on particle shape and surrounding environment. We study the effect of particle shape and substrate refractive index on the plasmonic resonances of silver nanoparticles and we systematically relate this to the efficiency of light scattering into a substrate. The light coupling spectra are dominated by Fano resonances for the corresponding dipolar and quadrupolar scattering modes. Varying the particle shape from spherical to cylindrical leads to large shifts in the Fano resonance for the dipolar mode, reducing the light incoupling integrated over the AM1.5 spectral range. Using a dielectric spacer layer, good light coupling is achieved for cylinders in the near-infrared. An asymmetric environment around the particles turns quadrupolar resonances into efficient radiators as well. PMID- 21643372 TI - Broadband UV-to-green photoconversion in V-doped lithium zinc silicate glasses and glass ceramics. AB - We report on photoluminescence of vanadium-doped lithium zinc silicate glasses and corresponding nanocrystalline Li2ZnSiO4 glass ceramics as broadband UV-to-VIS photoconverters. Depending on dopant concentration and synthesis conditions, VIS photoemission from [VO4]3 is centered at 550-590 nm and occurs over a bandwidth (FWHM) of ~250 nm. The corresponding excitation band covers the complete UV-B to UV-A spectral region. In as-melted glasses, the emission lifetime is about 34 MUs up to a nominal dopant concentration of 0.5 mol%. In the glass ceramic, it increases to about 45 MUs. For higher dopant concentration, a sharp drop in emission lifetime was observed, what is interpreted as a result of concentration quenching. Self-quenching is further promoted by energy transfer to V4+ centers (22Gt3). Partitioning of vanadium into V5+ and V4+ was examined by electron paramagnetic resonance and X-ray photoelectron spectroscopy. Suppression of V5+ reduction requires careful adjustment of the optical basicity of the host glass and/or synthesis conditions. PMID- 21643373 TI - Ultraviolet electroluminescence from hybrid inorganic/organic ZnO/GaN/poly(3 hexylthiophene) dual heterojunctions. AB - Based on hybrid inorganic/organic n-ZnO nanorods/p-GaN thin film/poly(3 hexylthiophene)(P3HT) dual heterojunctions, the light emitting diode (LED) emits ultraviolet (UV) radiation (370 nm - 400 nm) and the whole visible light (400 nm 700 nm) at the low injection current density. Meanwhile, under the high injection current density, the UV radiation overwhelmingly dominates the room-temperature electroluminescence spectra, exponentially increases with the injection current density and possesses a narrow full width at half maximum less than 16 nm. Comparing electroluminescence with photoluminescence spectra, an enormously enhanced transition probability of the UV luminescence in the electroluminescence spectra was found. The P3HT layer plays an essential role in helping the UV emission from p-GaN material because of its hole-conductive characteristic as well as the band alignment with respect to p-GaN. With our new finding, the result shown here may pave a new route for the development of high brightness LEDs derived from hybrid inorganic/organic heterojuctions. PMID- 21643374 TI - Fabrication of GaAs subwavelength structure (SWS) for solar cell applications. AB - We developed a novel GaAs subwavelength structure (SWS) as an antireflective layer for solar cell applications. The GaAs SWS patterns were fabricated by a combination of nanosphere lithography (NSL) and reactive ion etching (RIE). The shape and height of the GaAs SWS were controlled by the diameter of the SiO2 nanospheres and the etching time. Various GaAs SWS were characterized by the reflectance spectra. The average reflectance of the polished GaAs substrate from 200nm to 800nm was 35.1%. However, the average reflectance of the tapered GaAs SWS was reduced to 0.6% due to scattering and moth-eye effects. PMID- 21643375 TI - Eu2+-activated silicon-oxynitride Ca3Si2O4N2: a green-emitting phosphor for white LEDs. AB - The green-emitting phosphor Ca3Si2O4N2:Eu2+ was synthesized using a solid-state reaction. The luminescence properties, diffuse reflection spectrum, and thermal quenching were firstly studied, and a white light-emitting diode (wLED) was fabricated using the Eu2+-activated Ca3Si2O4N2 phosphor. Eu2+-doped Ca3Si2O4N2 exhibited a broad green emission band centered between 510 and 550 nm depending on the concentration of Eu2+. The optimal doping concentration of Eu2+ in Ca3Si2O4N2 was 1 mol%. The energy transfer between Eu2+ ions proceeds by an electric multipolar interaction mechanism, with a critical transfer distance of approximately 30.08 A. A wLED with an color-rendering index Ra of 88.25 at a correlated color temperature of 6029 K was obtained by combining a GaN-based n-UV LED (380 nm) with the blue-emitting BaMgAl10O17:Eu2+, green-emitting Ca3Si2O4N2:Eu2+, and red-emitting CaAlSiN3:Eu2+ phosphors. The results present Ca3Si2O4N2:Eu2+ as an attractive candidate for use as a conversion phosphor for wLED applications. PMID- 21643376 TI - Rigorous determination of stratospheric water vapor trends from MIPAS observations. AB - The trend of stratospheric water vapor as a function of latitude is estimated by the MIPAS measurements by means of a new method that uses the measurement space solution. The method uses all the information provided by the observations avoiding the artifacts introduced by the a priori information and by the interpolation to different vertical grids. The analysis provides very precise values of the trends that, however, are limited by a relatively large systematic error induced by the radiometric calibration error of the instrument. The results show in the five years from 2005 to 2009 a dependence on latitude of the stratospheric (from 37 to 53 km) water vapor trend with a positive value of (0.41 +/- 0.16)%yr-1 in the northern hemisphere and less than 0.16%yr-1 in the southern hemisphere. PMID- 21643377 TI - Enhancement of the power conversion efficiency by expanding the absorption spectrum with fluorescence layers. AB - The spectral response of Poly(3-hexylthiophene) (P3HT): 1-(3-methoxycarbonyl) propyl-1-phenyl-(6,6)C61 (PCBM) heterojunction film is between 350 nm and 650 nm, meaning that a lot of the sunlight is lost at ultraviolet and infrared regions. We fabricated solar cells by the attachment of a fluorescence layer which absorbs UV light, and emit visible light which will be re-used by P3HT, and thus the absorption spectrum is expanded. Since N,N'-bis(3-methylphenyl)-N,N'-bis(phenyl) benzidine (TPD) has high reflectance in the visible range, the usage of UV light will not manifest; when LiF is added as an antireflection layer, PCE was enhanced from 2.50% to 2.68%. PMID- 21643378 TI - Accurate estimation of the boundaries of a structured light pattern. AB - Depth recovery based on structured light using stripe patterns, especially for a region-based codec, demands accurate estimation of the true boundary of a light pattern captured on a camera image. This is because the accuracy of the estimated boundary has a direct impact on the accuracy of the depth recovery. However, recovering the true boundary of a light pattern is considered difficult due to the deformation incurred primarily by the texture-induced variation of the light reflectance at surface locales. Especially for heavily textured surfaces, the deformation of pattern boundaries becomes rather severe. We present here a novel (to the best of our knowledge) method to estimate the true boundaries of a light pattern that are severely deformed due to the heavy textures involved. First, a general formula that models the deformation of the projected light pattern at the imaging end is presented, taking into account not only the light reflectance variation but also the blurring along the optical passages. The local reflectance indices are then estimated by applying the model to two specially chosen reference projections, all-bright and all-dark. The estimated reflectance indices are to transform the edge-deformed, captured pattern signal into the edge corrected, canonical pattern signal. A canonical pattern implies the virtual pattern that would have resulted if there were neither the reflectance variation nor the blurring in imaging optics. Finally, we estimate the boundaries of a light pattern by intersecting the canonical form of a light pattern with that of its inverse pattern. The experimental results show that the proposed method results in significant improvements in the accuracy of the estimated boundaries under various adverse conditions. PMID- 21643379 TI - Traveling waves in two parallel infinite linear point-scatterer arrays. AB - Traveling waves in two coupled parallel infinite linear point-scatterer arrays are studied analytically for the first time to our knowledge. The two arrays are considered to be generally offset in the axial direction. It is found that slow quasi-even/odd supermodes are supported, as a result of the coupling-induced splitting of the modes of the single array, in direct analogy to standard optical waveguide couplers. Exactly even/odd supermodes are supported when the axial offset is zero. Mode splitting, dispersion curves, and coupling length are numerically investigated versus the inter-element spacing, the inter-array distance, and the axial offset. Potential applications of the concept are in directional optical couplers made of metallic or dielectric nanoparticle chains. PMID- 21643380 TI - M2-factor of truncated partially coherent beams propagating through atmospheric turbulence. AB - A method of studying the M2-factor of truncated partially coherent beams both in free space and in turbulence is proposed, i.e., the method of the window function being expanded into a finite sum of complex-valued Gaussian functions. Taking the Gaussian Schell-model (GSM) beam as a typical example of partially coherent beams, the analytical formula of the M2-factor of truncated GSM beams propagating through atmospheric turbulence is derived. It is shown that the M2-factor decreases as the truncation parameter delta and the coherence parameter alpha increase. However, the M2-factor in turbulence is more sensitive to delta than that in free space. On the other hand, the M2-factor of truncated partially coherent beams with smaller delta is more affected by turbulence. In addition, the effect of turbulence on the M2-factor of truncated GSM beams is less sensitive to the coherence parameter alpha than that of nontruncated GSM beams. PMID- 21643381 TI - Propagation of broadband gaussian Schell-model beams in the apertured fractional Fourier transformation systems. AB - On the basis of the fact that a hard-edged aperture function can be expressed as finite matrices with different weighting coefficients, we obtain the analytical formula for the propagation of the broadband gaussian Schell-model (BGSM) beam through the apertured fractional Fourier transformation (AFrFT) system. It is shown by numerical examples that the intensity distribution in the plane of a small fractional order is obviously influenced by the bandwidth when the BGSM beams propagate through the AFrFT system. Further extensions are also pointed out. PMID- 21643382 TI - Exact complex-wave reconstruction in digital holography. AB - We address the problem of exact complex-wave reconstruction in digital holography. We show that, by confining the object-wave modulation to one quadrant of the frequency domain, and by maintaining a reference-wave intensity higher than that of the object, one can achieve exact complex-wave reconstruction in the absence of noise. A feature of the proposed technique is that the zero-order artifact, which is commonly encountered in hologram reconstruction, can be completely suppressed in the absence of noise. The technique is noniterative and nonlinear. We also establish a connection between the reconstruction technique and homomorphic signal processing, which enables an interpretation of the technique from the perspective of deconvolution. Another key contribution of this paper is a direct link between the reconstruction technique and the two dimensional Hilbert transform formalism proposed by Hahn. We show that this connection leads to explicit Hilbert transform relations between the magnitude and phase of the complex wave encoded in the hologram. We also provide results on simulated as well as experimental data to validate the accuracy of the reconstruction technique. PMID- 21643383 TI - Wave field reconstruction from multiple plane intensity-only data: augmented lagrangian algorithm. AB - A complex-valued wave field is reconstructed from intensity-only measurements given at multiple observation planes parallel to the object plane. The phase retrieval algorithm is obtained from the constrained maximum likelihood approach provided that the additive noise is gaussian. The forward propagation from the object plane to the measurement plane is treated as a constraint in the proposed variational setting of reconstruction. The developed iterative algorithm is based on an augmented lagrangian technique. An advanced performance of the algorithm is demonstrated by numerical simulations. PMID- 21643384 TI - Transition between free-space Helmholtz equation solutions with plane sources and parabolic wave equation solutions. AB - The slowly varying envelope approximation is applied to the radiation problems of the Helmholtz equation with a planar single-layer and dipolar sources. The analyses of such problems provide procedures to recover solutions of the Helmholtz equation based on the evaluation of solutions of the parabolic wave equation at a given plane. Furthermore, the conditions that must be fulfilled to apply each procedure are also discussed. The relations to previous work are given as well. PMID- 21643385 TI - Correcting the diffusion approximation at the boundary. AB - The diffusion approximation to the radiative transport equation applies for light that has propagated deeply into an optically thick medium, such as biological tissue. It does not accurately model light near boundaries where measurements of scattered light are often taken. Here, we compute a correction to the diffusion approximation at the boundary. This correction requires only small modifications to the standard diffusion approximation used in biomedical optics. In particular, one needs only to compute the coefficients in the boundary condition for the diffusion approximation and an additive correction. We give explicit procedures for these computations. Using numerical results for the steady-state plane parallel slab problem, we show that this corrected diffusion approximation is a much better approximation than the standard diffusion approximation for modeling the reflectance and transmittance. PMID- 21643386 TI - Average spreading of a radial gaussian beam array in non-Kolmogorov turbulence. AB - The analytical expression for the rms beam width of the radial gaussian beam array propagating in non-Kolmogorov turbulence is derived, where the coherent combination is considered. The influences of the beam number, the generalized exponent, and the ring radius on the rms beam width are investigated. The results indicate that the rms beam width depends greatly on the generalized exponent and the beam number. Further, an optimum ring radius, which leads to a minimum beam width, is proved to exist within a certain traveling distance and the optimum ring radius increases when the beam number increases. PMID- 21643387 TI - Hybrid finite-element-boundary integral algorithm to solve the problem of scattering from a finite and infinite array of cavities with stratified dielectric coating. AB - This work presents a hybrid finite-element-boundary integral algorithm to solve the problem of scattering from a finite and infinite array of two-dimensional cavities engraved in a perfectly electric conducting screen covered with a stratified dielectric layer. The solution region is divided into interior regions containing the cavities and the region exterior to the cavities. The finite element formulation is applied only inside the interior regions to derive a linear system of equations associated with unknown field values. Using a two boundary formulation, the surface integral equation employing the grounded dielectric slab Green's function in the spatial domain is applied at the opening of the cavities as a boundary constraint to truncate the solution region. Placing the truncation boundary at the opening of the cavities and inside the dielectric layer results in a highly efficient solution in terms of computational resources, which makes the algorithm well suited for the optimization problems involving scattering from grating surfaces. The near fields are generated for an array of cavities with different dimensions and inhomogeneous fillings covered with dielectric layers. PMID- 21643388 TI - High-frequency homogenization for checkerboard structures: defect modes, ultrarefraction, and all-angle negative refraction. AB - The counterintuitive properties of photonic crystals, such as all-angle negative refraction (AANR) [J. Mod. Opt.34, 1589 (1987)] and high-directivity via ultrarefraction [Phys. Rev. Lett.89, 213902 (2002)], as well as localized defect modes, are known to be associated with anomalous dispersion near the edge of stop bands. We explore the implications of an asymptotic approach to uncover the underlying structure behind these phenomena. Conventional homogenization is widely assumed to be ineffective for modeling photonic crystals as it is limited to low frequencies when the wavelength is long relative to the microstructural length scales. Here a recently developed high-frequency homogenization (HFH) theory [Proc. R. Soc. Lond. A466, 2341 (2010)] is used to generate effective partial differential equations on a macroscale, which have the microscale embedded within them through averaged quantities, for checkerboard media. For physical applications, ultrarefraction is well described by an equivalent homogeneous medium with an effective refractive index given by the HFH procedure, the decay behavior of localized defect modes is characterized completely, and frequencies at which AANR occurs are all determined analytically. We illustrate our findings numerically with a finite-size checkerboard using finite elements, and we emphasize that conventional effective medium theory cannot handle such high frequencies. Finally, we look at light confinement effects in finite-size checkerboards behaving as open resonators when the condition for AANR is met [J. Phys. Condens. Matter 15, 6345 (2003)]. PMID- 21643389 TI - Compressive imaging: hybrid measurement basis design. AB - The inherent redundancy in natural scenes forms the basis of compressive imaging where the number of measurements is less than the dimensionality of the scene. The compressed sensing theory has shown that a purely random measurement basis can yield good reconstructions of sparse objects with relatively few measurements. However, additional prior knowledge about object statistics that is typically available is not exploited in the design of the random basis. In this work, we describe a hybrid measurement basis design that exploits the power spectral density statistics of natural scenes to minimize the reconstruction error by employing an optimal combination of a nonrandom basis and a purely random basis. Using simulation studies, we quantify the reconstruction error improvement achievable with the hybrid basis for a diverse set of natural images. We find that the hybrid basis can reduce the reconstruction error up to 77% or equivalently requires fewer measurements to achieve a desired reconstruction error compared to the purely random basis. It is also robust to varying levels of object sparsity and yields as much as 40% lower reconstruction error compared to the random basis in the presence of measurement noise. PMID- 21643390 TI - Metallic cylindrical focusing micromirrors with long axial focal depth or increased lateral resolution. AB - Using a general focal-length function, two-dimensional long-focal-depth (LFD) metallic cylindrical focusing micromirrors (MCFMs) are designed and the focal performance is systematically investigated based on rigorous electromagnetic theory and the boundary element method. For a positive preset focal depth, simulation results reveal that the designed MCFMs still possess an LFD property and high lateral resolution even when the f-number is reduced to f/0.3. On the other hand, through setting the preset focal depth to be negative, increased lateral resolution is obtained, compared with the conventional MCFM. In addition, under multiwavelength illumination, a large common LFD region is demonstrated for the designed LFD MCFMs, which is due to the intrinsic achromatic property of reflective systems. PMID- 21643391 TI - Manipulation of light with alpha transformation media. AB - A type of transformation media called alpha media is proposed by performing a direct transformation to the metric tensor of another kind of media, called seed media. Light rays in an alpha medium correlate to those in its seed medium through a simple displacement or rotation relation. Three types of commonly encountered anisotropic media are covered by the concept of alpha media: (1) media of slab shape, having continuous translational symmetry with respect to two Cartesian coordinate components; (2) media of cylindrical shape, having cylindrical rotational symmetry and continuous translational symmetry along the longitudinal direction; (3) media of spherical shape, having spherical rotational symmetry, with two principal axes along the symmetry directions, and with the material parameters in the same sign. Optical properties of such media can be effectively interpreted through recalling the properties of certain isotropic media, i.e., their seed media. Conversely, from simple isotropic media in which light trajectories are well known, one can design alpha media for manipulating light. Based on this fact, several optical devices, including frequency demultiplexers, beam splitters, focusing lenses, and radiation controllers, are designed and numerically verified. The famed invisibility cloak derived from a conventional coordinate transformation is revisited from the alpha media perspective. PMID- 21643392 TI - Light scattering by a coated infinite cylinder in an absorbing medium. AB - The scattering formulation for a coated infinite cylinder in an absorbing medium is presented in this paper. The cylinder is subjected to an arbitrarily polarized plane wave propagating in a general direction at the cylinder. The refractive index and magnetic permeability of the host medium, as well as those for the core and coating of the cylinder, can be real or complex. The scattering and extinction efficiencies and the scattering amplitudes are derived for both the near field and the far field. As the medium is absorbing, the "true" extinction and scattering efficiencies are derived based on the radiative energy outflow at the surface of the cylinder. The radiative efficiencies in the far field are denoted as "apparent" properties because they include absorption by the intervening medium. The influence of the refractive index and permeability of the host medium on the scattering properties of a coated cylinder is illustrated by numerical examples. PMID- 21643393 TI - Electromagnetic scattering by an infinite cylinder of material or metamaterial coating eccentrically a dielectric cylinder. AB - The electromagnetic scattering by an infinite cylinder of dielectric material or metamaterial, coating eccentrically another infinite dielectric cylinder, is treated in this work. The problem is solved using classical separation of variables techniques. No use is made of the translational addition theorem. For small eccentricities h = d/a(? 1), where d is the distance between the axes of the cylinders and a the radius of the outer cylinder, we use instead the cosine and the sine laws to satisfy the boundary conditions at the surface of the outer cylinder. Keeping terms up to the order h2 we finally obtain exact, closed-form expressions for the expansion coefficients g(1) and g(2) in the relation S(h) = S(0)[1 + g(1)h + g(2)h2 + O(h3)], giving the scattered field and the scattering cross sections of the problem, where S(0) corresponds to the coaxial geometry, with h = 0 (d = 0). Both polarizations are considered for normal incidence. Numerical results are given for various values of the parameters, corresponding to materials or metamaterials. Our method is an alternative of the one using the translational addition theorem in the case of small eccentricities h. PMID- 21643394 TI - Mie scattering in the time domain. Part 1. The role of surface waves. AB - We computed the Debye series p=1 and p=2 terms of the Mie scattered intensity as a function of scattering angle and delay time for a linearly polarized plane wave pulse incident on a spherical dielectric particle and physically interpreted the resulting numerical data. Radiation shed by electromagnetic surface waves plays a prominent role in the scattered intensity. We determined the surface wave phase and damping rate and studied the structure of the p=1,2 surface wave glory in the time domain. PMID- 21643395 TI - Mie scattering in the time domain. Part II. The role of diffraction. AB - The p=0 term of the Mie-Debye scattering amplitude contains the effects of external reflection and diffraction. We computed the reflected intensity in the time domain as a function of the scattering angle and delay time for a short electromagnetic pulse incident on a spherical particle and compared it to the predicted behavior in the forward-focusing region, the specular reflection region, and the glory region. We examined the physical consequences of three different approaches to the exact diffraction amplitude, and determined the signature of diffraction in the time domain. The external reflection surface wave amplitude gradually replaces the diffraction amplitude in the angular transition region between forward-focusing and the region of specular reflection. The details of this replacement were studied in the time domain. PMID- 21643396 TI - Nonredundant array of apertures to measure the spatial coherence in two dimensions with only one interferogram. AB - We propose to use a mask with a nonredundant array (NRA) of multiple apertures to measure spatial coherence in two dimensions. The spatial distribution of the apertures in the mask is made in such a way that we obtain a quasi-uniform sampling in the coherence domain. The spatial coherence is obtained by Fourier transform of the interferogram generated by the mask when it is illuminated by the light field under analysis. PMID- 21643398 TI - Linear systems formulation of scattering theory for rough surfaces with arbitrary incident and scattering angles. AB - Scattering effects from microtopographic surface roughness are merely nonparaxial diffraction phenomena resulting from random phase variations in the reflected or transmitted wavefront. Rayleigh-Rice, Beckmann-Kirchhoff. or Harvey-Shack surface scatter theories are commonly used to predict surface scatter effects. Smooth surface and/or paraxial approximations have severely limited the range of applicability of each of the above theoretical treatments. A recent linear systems formulation of nonparaxial scalar diffraction theory applied to surface scatter phenomena resulted first in an empirically modified Beckmann-Kirchhoff surface scatter model, then a generalized Harvey-Shack theory that produces accurate results for rougher surfaces than the Rayleigh-Rice theory and for larger incident and scattered angles than the classical Beckmann-Kirchhoff and the original Harvey-Shack theories. These new developments simplify the analysis and understanding of nonintuitive scattering behavior from rough surfaces illuminated at arbitrary incident angles. PMID- 21643397 TI - Image reconstruction in photoacoustic tomography involving layered acoustic media. AB - Photoacoustic tomography (PAT), also known as thermoacoustic or optoacoustic tomography, is a rapidly emerging biomedical imaging technique that combines optical image contrast with ultrasound detection principles. Most existing reconstruction algorithms for PAT assume the object of interest possesses homogeneous acoustic properties. The images produced by such algorithms can contain significant distortions and artifacts when the object's acoustic properties are spatially variant. In this work, we establish an image reconstruction formula for PAT applications in which a planar detection surface is employed and the to-be-imaged optical absorber is embedded in a known planar layered acoustic medium. The reconstruction formula is exact in a mathematical sense and accounts for multiple acoustic reflections between the layers of the medium. Computer-simulation studies are conducted to demonstrate and investigate the proposed method. PMID- 21643399 TI - Intensity-intensity fluctuations of stochastic fields produced upon weak scattering. AB - We derive expressions for the intensity-intensity correlations of a field produced on scattering of a random field governed by Gaussian statistics from a particle with a deterministic or a random refractive index distribution. Our results generalize the ones in [Opt. Lett. 35, 4000, 2010] to the case of an arbitrarily correlated incident field. We consider as specific examples the cases of a single plane wave and two partially correlated plane waves incident upon the scatterer. PMID- 21643400 TI - Channelized Hotelling observers for the assessment of volumetric imaging data sets. AB - Current clinical practice is rapidly moving in the direction of volumetric imaging. For two-dimensional (2D) images, task-based medical image quality is often assessed using numerical model observers. For three-dimensional (3D) images, however, these models have been little explored so far. In this work, first, two novel designs of a multislice channelized Hotelling observer (CHO) are proposed for the task of detecting 3D signals in 3D images. The novel designs are then compared and evaluated in a simulation study with five different CHO designs: a single-slice model, three multislice models, and a volumetric model. Four different random background statistics are considered, both gaussian (noncorrelated and correlated gaussian noise) and non-gaussian (lumpy and clustered lumpy backgrounds). Overall, the results show that the volumetric model outperforms the others, while the disparity between the models decreases for greater complexity of the detection task. Among the multislice models, the second proposed CHO could most closely approach the volumetric model, whereas the first new CHO seems to be least affected by the number of training samples. PMID- 21643401 TI - Scene-based nonuniformity correction algorithm based on interframe registration. AB - In this paper, we present a simple and effective scene-based nonuniformity correction (NUC) method for infrared focal plane arrays based on interframe registration. This method estimates the global translation between two adjacent frames and minimizes the mean square error between the two properly registered images to make any two detectors with the same scene produce the same output value. In this way, the accumulation of the registration error can be avoided and the NUC can be achieved. The advantages of the proposed algorithm lie in its low computational complexity and storage requirements and ability to capture temporal drifts in the nonuniformity parameters. The performance of the proposed technique is thoroughly studied with infrared image sequences with simulated nonuniformity and infrared imagery with real nonuniformity. It shows a significantly fast and reliable fixed-pattern noise reduction and obtains an effective frame-by-frame adaptive estimation of each detector's gain and offset. PMID- 21643402 TI - Scattering from cylinders using the two-dimensional vector plane wave spectrum. AB - The two-dimensional vector plane wave spectrum (VPWS) is scattered from parallel circular cylinders using a boundary value solution with the T-matrix formalism. The VPWS allows us to define the incident, two-dimensional electromagnetic field with an arbitrary distribution and polarization, including both radiative and evanescent components. Using the fast Fourier transform, we can quickly compute the multiple scattering of fields that have any particular functional or numerical form. We perform numerical simulations to investigate a grating of cylinders that is capable of converting an evanescent field into a set of propagating beams. The direction of propagation of each beam is directly related to a spatial frequency component of the incident evanescent field. PMID- 21643403 TI - Depth rendition of three-dimensional displays. AB - The third dimension in the reproduction of real scenes in three-dimensional displays is commonly subject to scale changes. The geometry of the situation is laid out, permitting the depth rendition of displays to be characterized and subjected to empirical examination. Psychophysical experiments are presented showing, even when geometrical deformations have been factored out, specific deviations from veridicality in observers' depth reports for stereograms of simple static patterns devoid of secondary cues. PMID- 21643404 TI - Boundary integral equation Neumann-to-Dirichlet map method for gratings in conical diffraction. AB - Boundary integral equation methods for diffraction gratings are particularly suitable for gratings with complicated material interfaces but are difficult to implement due to the quasi-periodic Green's function and the singular integrals at the corners. In this paper, the boundary integral equation Neumann-to Dirichlet map method for in-plane diffraction problems of gratings [Y. Wu and Y. Y. Lu, J. Opt. Soc. Am. A26, 2444 (2009)] is extended to conical diffraction problems. The method uses boundary integral equations to calculate the so-called Neumann-to-Dirichlet maps for homogeneous subdomains of the grating, so that the quasi-periodic Green's functions can be avoided. Since wave field components are coupled on material interfaces with the involvement of tangential derivatives, a least squares polynomial approximation technique is developed to evaluate tangential derivatives along these interfaces for conical diffraction problems. Numerical examples indicate that the method performs equally well for dielectric or metallic gratings. PMID- 21643405 TI - Group-index and resonant field enhancement in a symmetric double-sided grated waveguide. AB - A numerical study has been carried out by means of the Green's function method to explore possible performance improvements of a simple grated waveguide (GWg) by the variations of its grated structure. It is shown that a GWg featuring symmetric two-sided grated structure of 16 teeth with a 60 nm groove depth and having a symmetric refractive index profile with a relatively large contrast between the grated and ungrated layers is capable of delivering largely improved device performance compared to that achieved previously with a one-sided grating of 40 nm groove depth and asymmetric index profile. The improvement is characterized by a remarkable 8-fold and 15-fold increase in the group index and the maximum field intensity, respectively, at the first resonance wavelength above the upper band edge (shorter wavelength), while relatively less improvement is found at the first resonance wavelength below the lower band edge (longer wavelength). It is shown that more than 20% further improvement can be obtained by an appropriate shifting of the two innermost adjacent grating teeth in the case of the 40 nm groove depth. Apart from that, the result also reveals an interesting and remarkable correlation between the variations of the group index and the confined energy. PMID- 21643406 TI - Recognition and classification of red blood cells using digital holographic microscopy and data clustering with discriminant analysis. AB - We propose to apply statistical clustering algorithms on a three-dimensional profile of red blood cells (RBCs) obtained through digital holographic microscopy (DHM). We show that two classes of RBCs stored for 14 and 38 days can be effectively classified. Two-dimensional intensity images of these cells are virtually the same. DHM allows for measurement of the RBCs' biconcave profile, resulting in a discriminative dataset. Two statistical clustering algorithms are compared. A model-based clustering approach classifies the pixels of an RBC and recognizes the RBC as either new or old based. The K-means algorithm is applied to the four-dimensional feature vector extracted from the RBC profile. PMID- 21643407 TI - Synthesis of three-dimensional light fields with binary spatial light modulators. AB - Computation of a binary spatial light modulator (SLM) pattern that generates a desired light field is a challenging quantization problem for which several algorithms have been proposed, mainly for far-field or Fourier plane reconstructions. We study this problem assuming that the desired light field is synthesized within a volumetric region in the non-far-field range after free space propagation from the SLM plane. We use Fresnel and Rayleigh-Sommerfeld scalar diffraction theories for propagation of light. We show that, when the desired field is confined to a sufficiently narrow region of space, the ideal gray-level complex-valued SLM pattern generating it becomes sufficiently low pass (oversampled) so it can be successfully halftoned into a binary SLM pattern by solving two decoupled real-valued constrained halftoning problems. Our simulation results indicate that, when the synthesis region is considered, the binary SLM is indistinguishable from a lower resolution full complex gray-level SLM. In our approach, free space propagation related computations are done only once at the beginning, and the rest of the computation time is spent on carrying out standard image halftoning. PMID- 21643408 TI - Coupling of gaussian Schell-model beams into single-mode optical fibers. AB - We develop analytic equations that describe the mean and normalized variance of the coupling efficiency of gaussian Schell-model beams into single-mode optical fibers. Numerical methods and computer simulations are used to evaluate the accuracy of the various approximations used in this analysis, and, with some insight, empirical compensation is made for the identified shortcomings. The simulations make use of both speckled and nonspeckled beams by employing two different Monte Carlo methods to generate randomly drawn optical fields. While the analytic approximations break down in certain cases, the use of empirical compensation demonstrated accuracies of better than 5% for the mean coupling efficiency in all cases, and generally better than 40% for the coupling efficiency variance. By optimizing the compensation for particular beam characteristics, even higher accuracies can be achieved. PMID- 21643409 TI - Differential geometry of the ruled surfaces optically generated by mirror scanning devices: II. Generation of helicoids and hyperbolic paraboloids. AB - The theory developed in Part I of this study [Y. Li, "Differential geometry of the ruled surfaces optically generated by mirror-scanning devices. I. Intrinsic and extrinsic properties of the scan field," J. Opt. Soc. Am. A28, 667 (2011)] for the ruled surfaces optically generated by single-mirror scanning devices is extended to multimirror scanning systems for an investigation of optical generation of the well-known ruled surfaces, such as helicoid, Plucker's conoid, and hyperbolic paraboloid. PMID- 21643410 TI - Airy pulsed beams. AB - The Airy beam (AiB) has attracted a lot of attention recently because of its intriguing features; the most distinctive ones are the propagation along curved trajectories in free space and the weak diffraction. We have previously shown that the AiB is, in fact, a caustic of the rays that radiate from the tail of the Airy function aperture distribution. Here we derive a class of ultra wideband Airy pulsed beams (AiPBs), which are the extension of the AiB into the time domain. We introduce a frequency scaling of the initial aperture field that renders the ray skeleton of the field, including the caustic, frequency independent, thus ensuring that all the frequency components propagate along the same curved trajectory and that the AiPB does not disperse. The resulting AiPB preserves the intriguing features of the time-harmonic AiB discussed above. An exact closed-form solution for the AiPB is derived using the spectral theory of transients. We also derive wavefront approximations for the field in the time window around the pulse arrival, which are valid uniformly in the vicinity of the caustic. These approximations are based on the so-called uniform geometrical optics, which is extended here to the time domain. PMID- 21643411 TI - Principal angles and principal azimuths of frustrated total internal reflection and optical tunneling by an embedded low-index thin film. AB - The condition for obtaining a differential (or ellipsometric) quarter-wave retardation when p- and s-polarized light of wavelength lambda experience frustrated total internal reflection (FTIR) and optical tunneling at angles of incidence phi >= the critical angle by a transparent thin film (medium 1) of low refractive index n1 and uniform thickness d, which is embedded in a transparent bulk medium 0 of high refractive index n0 takes the simple form: -tanh2 x = tan deltap tan deltas, in which x = 2pin1(d/lambda)(N2sin2phi - 1)(1/2), N = n0/n1, and deltap, deltas are 01 interface Fresnel reflection phase shifts for the p and s polarizations. From this condition, the ranges of the principal angle and normalized film thickness d/lambda are obtained explicitly. At a given principal angle, the associated principal azimuths psir, psit in reflection and transmission are determined by tan2psir = -sin 2deltas/sin 2deltap and tan2psit = -tan deltap/tan deltas, respectively. At a unique principal angle phie given by sin2phie = 2/(N2 + 1), psir = psit = 45 degrees and linear-to-circular polarization conversion is achieved upon FTIR and optical tunneling simultaneously. The intensity transmittances of p- and s-polarized light at any principal angle are given by taup = tan deltap/tan (deltap - deltas) and taus = tan deltas/tan (deltap - deltas), respectively. The efficiency of linear-to circular polarization conversion in optical tunneling is maximum at phie. PMID- 21643412 TI - Experimental and theoretical investigation of the pixel saturation effect in digital holography. AB - This paper presents an experimental investigation and an analytical modeling of the nonlinear pixel saturation effect in digital off-axis holography. The theoretical analysis is based on a semiempirical modeling and supported by the experimental analysis. Taking into account the nonlinearity of the phenomenon, an exponential law for the high-order harmonic amplitude is proposed and validated by the experimental results. The conclusion of this analysis is that the saturation effect can be described by the use of a linear operator that involves autoconvolution of the initial object wave, even though the saturation phenomenon is nonlinear. PMID- 21643413 TI - Real-time convolution method for generating light diffusion profiles of layered turbid media. AB - In this paper we present a technique to obtain a diffusion profile of layered turbid media in real time by using the quasi fast Hankel transform (QFHT) and the latest graphics processing unit technique. We apply the QFHT to convolve the diffusion profiles of each layer so as to dramatically reduce the time for the convolution step while maintaining the accuracy. In addition, we also introduce an accelerated technique to generate individual discrete diffusion profiles for each layer through parallel processing. The proposed method is 2 orders of magnitude faster than the existing method, and we validate its efficiency by comparing it with Monte Carlo simulation and another relevant methods. PMID- 21643414 TI - Scale transformation of Maxwell's equations and scattering by an elliptic cylinder. AB - A scale transformation that converts an ellipse into a circle has been suggested in the literature as a method for eliminating the need to evaluate the conventional Mathieu function solution for scattering by an elliptic cylinder. This suggestion is tested by examining the wave equation in the scaled coordinate system and by evaluating the scattering from a thin ellipse for conditions where it is expected that an approximate solution can be obtained using the scalar theory single-slit approximation. It is found that, for a plane electromagnetic wave normally incident on a thin perfectly conducting ellipse, the position of the first minimum in the diffraction pattern, relative to the central axis, differs by approximately a factor of 7 between the single-slit and the scaled theory approach to the problem. The examination of the scaled wave equation and the scattering calculation suggests that, because the scale transformation generates an anisotropic medium, the use of a uniform medium solution in the scaled coordinate system is not appropriate. PMID- 21643415 TI - Digital holographic reconstruction of a local object field using an adjustable magnification. AB - In the research of digital holography, this paper presents a numerical method using an adjustable magnification for local object field reconstruction together with experiment verification. The method first designs a spherical wave according to the given magnification to illuminate the digital hologram, then through a Fourier transform of diffraction, it calculates the reconstructed image plane. Afterward, a filtering window is set in the image plane to extract the image of the local object field, and then the object field reached hologram plane is formed using diffraction's inverse operation. Finally, the object field is reconstructed through diffraction's angular spectrum theory. PMID- 21643416 TI - Quantitative Carre differential interference contrast microscopy to assess phase and amplitude. AB - We present a method of using an unmodified differential interference contrast microscope to acquire quantitative information on scatter and absorption of thin tissue samples. A simple calibration process is discussed that uses a standard optical wedge. Subsequently, we present a phase-stepping procedure for acquiring phase gradient information exclusive of absorption effects. The procedure results in two-dimensional maps of the local angular (polar and azimuthal) ray deviation. We demonstrate the calibration process, discuss details of the phase-stepping algorithm, and present representative results for a porcine skin sample. PMID- 21643417 TI - Wigner distribution function of an Airy beam. AB - We study the Wigner distribution function (WDF) of an Airy beam. The analytical expression of the WDF of an Airy beam is obtained. Numerical and graphical results of the WDF of an Airy beam provide an intuitive picture to explain the intriguing features of an Airy beam, such as weak diffraction, curved propagation, and self-healing. Our results confirm that these novel properties of an Airy beam are attributed to the continuum of sideways contributions to the field. PMID- 21643418 TI - Refracting the k-function: Stavroudis's solution to the eikonal equation for multielement optical systems. AB - The k-function of Stavroudis describes a solution of the eikonal equation in a region of constant refractive index. Given the k-function describing the optical field in one region of space, and given a prescribed refractive or reflective boundary, we construct the k-function for the refracted or reflected field. This procedure, which Stavroudis calls refracting the k-function, can be repeated any number of times, and therefore extends the usefulness of the k-function formalism to multielement optical systems. As examples, we present an analytic solution for the k-function, wavefronts, and caustics generated by a biconvex thick lens illuminated by a plane wave propagating parallel to the symmetry axis, and numerical results for off-axis plane-wave illumination of a two-mirror telescope. PMID- 21643419 TI - Luminance-based specular gloss characterization. AB - Gloss is a feature of visual appearance that arises from the directionally selective reflection of light incident on a surface. Especially when a distinct reflected image is perceptible, the luminance distribution of the illumination scene above the sample can strongly influence the gloss perception. For this reason, industrial glossmeters do not provide a satisfactory gloss estimation of high-gloss surfaces. In this study, the influence of the conditions of illumination on specular gloss perception was examined through a magnitude estimation experiment in which 10 observers took part. A light booth with two light sources was utilized: the mirror image of only one source being visible in reflection by the observer. The luminance of both the reflected image and the adjacent sample surface could be independently varied by separate adjustment of the intensity of the two light sources. A psychophysical scaling function was derived, relating the visual gloss estimations to the measured luminance of both the reflected image and the off-specular sample background. The generalization error of the model was estimated through a validation experiment performed by 10 other observers. In result, a metric including both surface and illumination properties is provided. Based on this metric, improved gloss evaluation methods and instruments could be developed. PMID- 21643420 TI - Characterisation of the PTEN inhibitor VO-OHpic. AB - PTEN (phosphatase and tensin homologue deleted on chromosome 10) is a phosphatidylinositol triphosphate 3-phosphatase that counteracts phosphoinositide 3-kinases and has subsequently been implied as a valuable drug target for diabetes and cancer. Recently, we demonstrated that VO-OHpic is an extremely potent inhibitor of PTEN with nanomolar affinity in vitro and in vivo. Given the importance of this inhibitor for future drug design and development, its mode of action needed to be elucidated. It was discovered that inhibition of recombinant PTEN by VO-OHpic is fully reversible. Both K(m) and V(max) are affected by VO OHpic, demonstrating a noncompetitive inhibition of PTEN. The inhibition constants K(ic) and K(iu) were determined to be 27 +/- 6 and 45 +/- 11 nM, respectively. Using the artificial phosphatase substrate 3-O-methylfluorescein phosphate (OMFP) or the physiological substrate phosphatidylinositol 3,4,5 triphosphate (PIP(3)) comparable parameters were obtained suggesting that OMFP is a suitable substrate for PTEN inhibition studies and PTEN drug screening. PMID- 21643421 TI - HIV Prevalence Comparison Between Antenatal Sentinel Surveillance and Demographic and Health Survey in Rwanda. AB - OBJECTIVE: To compare HIV prevalence from antenatal surveillance to that of the demographic and health survey (DHS), and to identify factors determining the difference of HIV prevalence between women recruited in these two surveys in Rwanda in 2005. METHODS: Comparative cross-sectional study of HIV prevalence and socio-demographic factors collected by the antenatal survey in 13,745 pregnant women, seen in 30 health centres located throughout the country and those collected by the DHS among 5641 women, aged 15-49 years living in households located throughout the country. Log-binomial regression and direct standardization were used to estimate and compare HIV prevalence between the two surveys. RESULTS: HIV prevalence in the antenatal survey was slightly higher than that in DHS women (4.1% versus 3.6% p=0.103). Socio-demographic characteristics were differently distributed between the two populations. Whereas, 59%, 93%, 53% of pregnant women were aged 20-29 years, married or cohabiting and living in rural areas respectively, the corresponding proportions among DHS women were 35%, 48% and 83% (p<0.001). Simultaneous standardization of antenatal prevalence according to the distribution of socio-demographic characteristics in the DHS gave an overall HIV prevalence estimate of 3.6%, similar to the prevalence measured among DHS women. CONCLUSIONS: HIV prevalence in the antenatal survey overestimated that among women of the general population in Rwanda in 2005. This overestimation could be corrected by standardization of antenatal prevalence according to the distribution of age, geographical area, marital status, parity, and education, in the general population. PMID- 21643422 TI - A Multicenter, Open Labeled, Randomized, Phase III Study Comparing Lopinavir/Ritonavir Plus Atazanavir to Lopinavir/Ritonavir Plus Zidovudine and Lamivudine in Naive HIV-1-Infected Patients: 48-Week Analysis of the LORAN Trial. AB - OBJECTIVE: The primary aim of the study was to compare the metabolic side effects of a nucleoside analogue-containing regimen with a nucleoside analogue-sparing double protease inhibitor regimen. A secondary goal was to test for efficacy of a double-PI regimen. DESIGN: Multicenter, randomized, open-label, phase III clinical trial. SUBJECTS: Adult HIV-1-infected individuals naive to antiretroviral therapy with viral load above 400 HIV-RNA copies/ml were randomized (1:1) to either 400 mg lopinavir /100 mg ritonavir (LPV/r) BID plus 150 mg lamivudine/300 mg zidovudine (CBV) BID versus LPV/r BID plus 300 mg atazanavir (ATV) QD. Main outcome measure was the virologic failure in both groups, defined as viral load >=50 copies/ml at week 48. RESULTS: In the CBV/LPV/r-arm, 29 out of 35 patients [(83%; 95% confidence interval (CI) 66.9 92.2%] and 18 out of 40 patients (45%; 95% CI 29.7-61.5%) in the ATV/LPV/r-arm had a HIV-RNA level <50 copies/ml at week 48. The intent-to-treat analysis revealed inferior virologic response in the ATV/LPV/r arm (Chi-Q and Fisher's Exact Test p<0.001) and resulted in premature termination of the trial. Eleven patients in the ATV/LPV/r-arm discontinued therapy because of virological failure. These failures mostly presented with low level replication (<1,000 copies/ml). Increases in CD4 cell counts was significantly more rapid in the ATV/LPV/r arm (p=0.02), but comparable at week 48. CONCLUSIONS: ATV/LPV/r had less virologic efficacy than the conventional RTI-based regimen and resulted in a high virological failure rate with low level replication. PMID- 21643423 TI - Knowledge of Pregnant Women on Mother-to-Child Transmission of HIV in Yaounde. AB - INTRODUCTION: Mother-to-child transmission of HIV is a major public health problem in Cameroon. The aim of this study was to assess knowledge of pregnant women on HIV transmission and prevention, particularly the four pillars of mother to-child transmission. MATERIALS AND METHODS: This was a descriptive cross sectional study from October 2008 to January 2009 at the Yaounde Gynaecology Obstetrics and Pediatric Hospital. All women presenting at the Ante-Natal Care (ANC) Clinic for the first time were included in the study after obtaining a verbal informed consent. RESULTS: The ages of the 260 women included in the study ranged from 15 to 42 years. Almost 99% (257/260) of the women interviewed had heard about HIV. Respectively, 80.5% (209/260), 89.3% (232/260) and 81.2% (211/260) of the women cited pregnancy, delivery, and breastfeeding as risk periods for HIV transmission from mother to child. Use of the male condom, the female condom, abstinence, and faithfulness to a single partner were considered as effective methods of HIV prevention by 73% (190/260), 76% (198/260), 88% (229/260) and 46% (120/260) of respondents respectively. About 79% (64/81) of participants with higher education considered HIV infection to be contagious as opposed to 45.5% (5/11) of women with no formal education (P = 0.008). CONCLUSION: Our data suggest that women have some good knowledge on the Prevention of Mother-To-Child Transmission (PMTCT) of HIV. Nevertheless, improving the formal educational level of these women may contribute to a further reduction of HIV transmission. PMID- 21643424 TI - In-vitro Strain and Modulus Measurements in Porcine Cervical Lymph Nodes. AB - Cervical lymph nodes are common sites of metastatic involvement in head and neck cancers. These lymph nodes are superficially located and palpation is a common practice for assessing nodal hardness and staging cancer which is, however, too subjective and with limited accuracy. In this study, the mechanical properties of pig lymph node tissues were investigated using ultrasound elastography and indentation test. Lymph nodes were excised from fresh pork pieces and embedded in an agar-gelatin phantom for strain imaging by elastography. A strain ratio reflecting the strain contrast of lymph node over agar-gelatin phantom was used to assess the elasticity of the lymph node. A cutting device was then custom designed to slice the phantom into uniform slices for indentation test. The measurements revealed that there were significant differences in both the strain ratio and Young's modulus between the peripheral and middle regions of the lymph nodes (both p < 0.05); however, the results appeared contradictory. Correlation between the results of the two measurements (modulus ratio vs. inversed strain ratio) showed their association was moderate for both the peripheral and middle regions (R(2) = 0.437 and 0.424 respectively). As the tests were only performed on normal lymph nodes, comparison in stiffness between healthy and abnormal lymph nodes could not be made. Future studies should be conducted to quantify the stiffness change in abnormal lymph nodes. PMID- 21643425 TI - Particle Trajectories and Agglomeration/Accumulation in Branching Arteries subjected to Orbital Atherectomy. AB - BACKGROUND: The transport of particles in surrogate and actual arterial geometries has been investigated synergistically by experimentation and numerical simulation. The motivating application for this work is orbital atherectomy which spawns a particle cloud in the process of debulking plaque from arterial walls. METHODS: Paired simulations and experiments were performed to prove the capability of the simulation model to predict both fluid and particle motions in branched arterial geometries. The verified model was then employed to predict the pattern of fluid flow in an actual multi-branched arterial geometry, including the flowrates passing through each of the individual branches. These predictions are in very good agreement with experimental data. Focus was then shifted to the issues of particle agglomeration within the flowing fluid and particle accumulation on the vessel walls. Once again, a synergistic approach was used. Flow visualization was employed to track the particle motions and to identify possible particle agglomeration within the fluid. RESULTS AND CONCLUSIONS: Accumulation of particles on walls was identified by measuring size distributions of effluent and residue within the artery. Scanning Electron Microscopy (SEM) evaluation showed evidence of a size-based sorting as the particles passed through vessels. It was found that plaque-facsimile particles resisted particle particle agglomeration. They also did not accumulate to the wall of the facsimile artery. In addition, simulations showed that if particle-wall accumulation were to occur, it would be limited to very small regions in the artery branches. PMID- 21643426 TI - Pseudophakic cystoid macular edema associated with extrafoveal vitreoretinal traction. AB - PURPOSE: To describe an association between extrafoveal vitreoretinal traction and intractable chronic pseudophakic cystoid macular edema (CME) by the use of optical coherence tomography (OCT). METHODS: In a retrospective case series study, charts and OCT findings of patients who had postoperative recalcitrant pseudophakic CME for at least 6 months and vitreoretinal traction membranes were analyzed. Excluded were eyes that either had another vitreoretinopathy that could affect the analysis or had undergone an intravitreal intervention. RESULTS: Three eyes (three patients) with macular edema following uneventful cataract surgery were detected to be associated with multifocal extrafoveal vitreoretinal traction sites in each. Retinal edema that was underlying each of the traction sites in all eyes was in continuum in at least one site per eye with the central macular edema, thus manifesting as diffuse macular edema. CONCLUSION: Chronic pseudophakic macular edema may be related to extrafoveal vitreoretinal traction. PMID- 21643427 TI - Long-Term Outcomes of Selective Laser Trabeculoplasty (SLT) Treatment. AB - PURPOSE: Glaucoma is a progressive optic neuropathy that may lead to blindness. Reducing intraocular pressure (IOP) seems to be the only treatment that slows progression in glaucoma. IOP can be decreased by pharmaceutical treatment, laser treatment or surgery. The aim of the present study was to assess the long-term efficacy of selective laser trabeculoplasty (SLT) treatment. METHODS: Retrospective chart review of eyes that underwent SLT between 1 January 2005 and 31 December 2005. The primary outcome measure was time to failure after SLT treatment. Failure after SLT was defined as any one or more of the following: change in the medical treatment, performance of a further SLT treatment, the patient being sent for surgery. All patients were treated over 90 degrees with SLT. RESULTS: 120 eyes of 120 patients were identified. The average time to failure after SLT was 18 months. The success rate after 12 months was 62%, after 24 months 34%, after 36 months 28% and after 48 months 24%. CONCLUSIONS: The long term effects of SLT when eyes were treated over 90 degrees seem to be low. The authors recommend treating patients over 180 degrees , as has traditionally been done. We suggest that this will improve the long-term results. PMID- 21643428 TI - Microsporidial keratoconjunctivitis in the tropics: a case series. AB - PURPOSE: To present a series of microsporidial keratoconjunctivitis in 24 eyes. METHODS: Retrospective non-comparative observational case series. Medical records were retrieved and individuals evaluated based on symptoms, risk factors, visual acuity, slit lamp biomicroscopy and pathological examination of cornea epithelial scrapings. Demographic features, clinical course, predisposing factors, microbiological profile, treatment, final clinical outcome and visual acuity were recorded. RESULTS: Of the 22 patients, 90.9% were men, with a mean age of 30.3 years (range 15 - 76 years). Two (9.1%) had bilateral involvement, 15 (68.2%) were non-contact lens users, 17 (77.3%) reported contamination with mud within 2 weeks (mean 6.8 days) of onset of symptoms. All patients presented with conjunctivitis and coarse, multifocal, punctate epithelial keratitis. Two out of 24 eyes (8.3%) had anterior stromal infiltrates, while 8 (33.3%) had anterior uveitis. Microsporidial spores were identified on modified trichrome staining of corneal epithelial scrapes in all eyes. All eyes were treated with epithelial debridement, topical fluoroquinolone and hexamidine diisethionate, 7 (31.8%) patients received oral albendazole, and all eyes with anterior uveitis received topical steroids. All cases resolved without visually significant sequelae. CONCLUSION: Microsporidial keratoconjunctivitis occurred mainly in males, is usually unilateral, presents as conjunctivitis and coarse, multifocal, punctate epithelial keratitis, and may incite anterior uveitis. Soil contamination is an important risk factor. Treatment with debridement, fluoroquinolones, hexamidine diisethionate with or without systemic albendazole is effective, with steroids reserved for any associated anterior uveitis. PMID- 21643429 TI - Determination of in vitro bioaccessibility of Pb, As, Cd and Hg in selected traditional Indian medicines. AB - In vitro bioaccessibility of Pb, As, Cd and Hg in five traditional Indian medicine samples was measured as a determinant of bioavailability. The method is based on simulation of human digestion in the passage of material from the gastric to intestinal portions of the gastrointestinal tract. Total concentration and concentration in extracts from gastric and intestinal phases were analyzed for Pb, As and Cd by dynamic reaction cell inductively coupled plasma mass spectrometry (DRC-ICP-MS) and for Hg by direct mercury analyzer (DMA). Total lead ranged from 1.9 to 36000 ug g(-1). In each of the samples bioaccessibility of lead was significantly higher (range 28-88%) in the gastric phase than in the intestinal phase (range 1.4-75.4%). Only Ekangvir Ras had measurable arsenic (304 ug g(-1)). Its bioaccessibility in the gastric phase and intestinal phase was 82.6% and 78.1%, respectively. Only Ayu-Nephro-Tone had measurable cadmium (14.4 ug g(-1)). Its bioaccessibility in the gastric phase and intestinal phase was 80.5% and 2.2%, respectively. Three samples had measurable mercury (range 37 ug g(-1)-10000 ug g(-1)). Mercury in these samples was not bioaccessible. For the samples with measurable amount of metal, the estimated daily amount of bioaccessible (EDAB) metal was calculated. When compared with the most liberal published safety guideline, EDAB-Pb in Mahayograj Guggulu and Ekangvir Ras were 37 and 45 fold greater. When compared with the most conservative published safety guideline, all samples had higher EDAB-Pb or EDAB-As than the suggested limits. The EDAB-Cd and EDAB-Hg were acceptably below published safety limits. PMID- 21643430 TI - Birth Outcomes, Lifetime Alcohol Dependence and Cognition in Middle Adulthood. AB - Prenatal exposure to alcohol is associated with cognitive abnormalities that persist throughout the lifespan and are also often a focus of studies examining cognitive outcomes associated with excessive alcohol use by an individual. This study examined the effect of birth outcomes consistent with fetal alcohol exposure on associations between lifetime alcohol dependence and cognition in middle adulthood. The sample was comprised of 315 adult adoptees ranging in age from 31 to 64 years (SD = 7.20). Facial morphology, pre-morbid cognition, and current cognition were assessed. Birth parent behaviors and birth outcomes (e.g., birthweight, gestational age) were obtained from adoption agency records. Lifetime alcohol dependence was determined from the Semi-Structured Assessment of the Genetics of Alcoholism - II. Univariate associations showed significantly poorer pre-morbid and current cognition when birth parent problems, short palpebral fissures, and thin upper lips were present. Lifetime alcohol dependence was associated with lower perceptional organization, processing speed and working memory. Multivariate analyses demonstrated continued significance suggesting unique contributions of each to cognition. Evaluating the possible role of fetal alcohol exposure within studies on alcoholism can only further improve the treatment and prevention of alcohol-related problems by isolating those cognitive outcomes uniquely attributable to an individual's consumption of alcohol. PMID- 21643431 TI - Hangover Predicts Residual Alcohol Effects on Psychomotor Vigilance the Morning After Intoxication. AB - OBJECTIVES: Both hangover and performance deficits have been documented the day after drinking to intoxication after breath alcohol concentration (BrAC) has returned to near zero. But few studies have examined the relationship between hangover and post-intoxication performance. METHOD: We performed secondary analyses of data from a previously reported controlled cross-over laboratory study to assess the relationship of hangover incidence and severity to sustained attention/reaction time the morning after drinking to about 0.11 g% BrAC. Relationships were investigated while controlling for gender, type of alcoholic beverage (bourbon or vodka), and neurocognitive performance after placebo. RESULTS: Hangover severity and neurocognitive performance were significantly correlated. Participants reporting stronger hangover were more impaired than those reporting little or no hangover. Comparing any to no hangover showed a trend in the same direction of effect. CONCLUSIONS: More intense hangover may indicate less fitness for duty in workers in certain safety-sensitive occupations, with implications for occupational alcohol policies. PMID- 21643432 TI - POVERTY, INFANT MORTALITY, AND HOMICIDE RATES IN CROSS-NATIONAL PERPSECTIVE: ASSESSMENTS OF CRITERION AND CONSTRUCT VALIDITY. AB - This paper assesses the extent to which the infant mortality rate might be treated as a "proxy" for poverty in research on cross-national variation in homicide rates. We have assembled a pooled, cross-sectional time-series dataset for 16 advanced nations over the 1993-2000 period that includes standard measures of infant mortality and homicide and also contains information on two commonly used "income-based" poverty measures: a measure intended to reflect "absolute" deprivation and a measure intended to reflect "relative" deprivation. With these data, we are able to assess the criterion validity of the infant mortality rate with reference to the two income-based poverty measures. We are also able to estimate the effects of the various indicators of disadvantage on homicide rates in regression models, thereby assessing construct validity. The results reveal that the infant mortality rate is more strongly correlated with "relative poverty" than with "absolute poverty," although much unexplained variance remains. In the regression models, the measure of infant mortality and the relative poverty measure yield significant positive effects on homicide rates, while the absolute poverty measure does not exhibit any significant effects. Our analyses suggest that it would be premature to dismiss relative deprivation in cross-national research on homicide, and that disadvantage is best conceptualized and measured as a multidimensional construct. PMID- 21643433 TI - Land Cover Classification in a Complex Urban-Rural Landscape with Quickbird Imagery. AB - High spatial resolution images have been increasingly used for urban land use/cover classification, but the high spectral variation within the same land cover, the spectral confusion among different land covers, and the shadow problem often lead to poor classification performance based on the traditional per-pixel spectral-based classification methods. This paper explores approaches to improve urban land cover classification with Quickbird imagery. Traditional per-pixel spectral-based supervised classification, incorporation of textural images and multispectral images, spectral-spatial classifier, and segmentation-based classification are examined in a relatively new developing urban landscape, Lucas do Rio Verde in Mato Grosso State, Brazil. This research shows that use of spatial information during the image classification procedure, either through the integrated use of textural and spectral images or through the use of segmentation based classification method, can significantly improve land cover classification performance. PMID- 21643434 TI - Impervious surface mapping with Quickbird imagery. AB - This research selects two study areas with different urban developments, sizes, and spatial patterns to explore the suitable methods for mapping impervious surface distribution using Quickbird imagery. The selected methods include per pixel based supervised classification, segmentation-based classification, and a hybrid method. A comparative analysis of the results indicates that per-pixel based supervised classification produces a large number of "salt-and-pepper" pixels, and segmentation based methods can significantly reduce this problem. However, neither method can effectively solve the spectral confusion of impervious surfaces with water/wetland and bare soils and the impacts of shadows. In order to accurately map impervious surface distribution from Quickbird images, manual editing is necessary and may be the only way to extract impervious surfaces from the confused land covers and the shadow problem. This research indicates that the hybrid method consisting of thresholding techniques, unsupervised classification and limited manual editing provides the best performance. PMID- 21643435 TI - The Relation between Profiles of Leisure Activity Participation and Substance Use among South African Youth. AB - A large body of research indicates that leisure activity participation is associated with substance use for American adolescents, and that leisure may be an important context of substance use prevention. It is important to begin to apply what we have learned with American youth and extend the knowledge base in other countries with significant adolescent risk behavior. The current study examined the association between leisure activity participation and substance use among a predominately Colored sample of 3497 South African 8(th) graders. Males' activity participation was characterized by five leisure activity profiles (Uninvolved; Sports and Volunteer; Mixed: Recreation and Hobbies; Mixed: Artistic; Highly Involved), whereas females' activity participation was characterized by four leisure activity profiles (Uninvolved; Uninvolved but Social; Mixed; Highly Involved). Leisure activity profiles were significantly associated with past-month alcohol, tobacco, and marijuana use. PMID- 21643436 TI - Evaluating Functional Autocorrelation within Spatially Distributed Neural Processing Networks. AB - Data-driven statistical approaches, such as cluster analysis or independent component analysis, applied to in vivo functional neuroimaging data help to identify neural processing networks that exhibit similar task-related or restingstate patterns of activity. Ideally, the measured brain activity for voxels within such networks should exhibit high autocorrelation. An important limitation is that the algorithms do not typically quantify or statistically test the strength or nature of the within-network relatedness between voxels. To extend the results given by such data-driven analyses, we propose the use of Moran's I statistic to measure the degree of functional autocorrelation within identified neural processing networks and to evaluate the statistical significance of the observed associations. We adapt the conventional definition of Moran's I, for applicability to neuroimaging analyses, by defining the global autocorrelation index using network-based neighborhoods. Also, we compute network specific contributions to the overall autocorrelation. We present results from a bootstrap analysis that provide empirical support for the use of our hypothesis testing framework. We illustrate our methodology using positron emission tomography (PET) data from a study that examines the neural representation of working memory among individuals with schizophrenia and functional magnetic resonance imaging (fMRI) data from a study of depression. PMID- 21643437 TI - In vitro remineralization of human and bovine white-spot enamel lesions by NaF dentifrices: A pilot study. AB - The aim of this feasibility study was to evaluate the in vitro remineralization effects of four dentifrice systems using microhardness and fluoride uptake analyses. In vitro testing for the potential remineralization of the white-spot lesions in bovine and human enamel was performed using a 10-day pH cycling model. The study involved the following NaF silica-based dentifrices: 1) placebo (0 ppm F), 2) 500 ppm F, 3) 1150 ppm F, and 4) 500 ppm F plus functionalized tricalcium phosphate (fTCP). Each day consisted of four two-minute treatments, one four-hour acid challenge (pH = 5.0), and immersion in artificial saliva (pH = 7.0) between these events. After cycling, specimens were analyzed for surface microhardness (SMH), enamel fluoride uptake (EFU), and cross-sectional microhardness (CSM). Statistical analyses revealed significant differences (ANOVA, LSD, p<0.05) among the four groups, with the placebo and 500 ppm F dentifrices providing significantly less remineralization relative to the 1150 ppm F and 500 ppm F plus fTCP dentifrices. Notably, while CSM measurements for both enamel types generated similar profiles for the four groups, SMH and EFU revealed human enamel was more sensitive to the 500 ppm F dentifrice groups compared to bovine enamel. This apparent sensitivity may be due to the inherent structural differences between the two substrates. PMID- 21643438 TI - High-Quality Protein Crystal Growth of Mouse Lipocalin-Type Prostaglandin D Synthase in Microgravity. AB - Lipocalin-type prostaglandin (PG) D synthase (L-PGDS) catalyzes the isomerization of PGH(2) to PGD(2) and is involved in the regulation of pain and of nonrapid eye movement sleep and the differentiation of male genital organs and adipocytes, etc. L-PGDS is secreted into various body fluids and binds various lipophilic compounds with high affinities, acting also as an extracellular transporter. Mouse L-PGDS with a C65A mutation was previously crystallized with citrate or malonate as a precipitant, and the X-ray crystallographic structure was determined at 2.0 A resolution. To obtain high-quality crystals, we tried, unsuccessfully, to crystallize the C65A mutant in microgravity under the same conditions used in the previous study. After further purifying the protein and changing the precipitant to polyethylene glycol (PEG) 8000, high-quality crystals were grown in microgravity. The precipitant solution was 40% (w/v) PEG 8000, 100 mM sodium chloride, and 100 mM HEPES-NaOH (pH 7.0). Crystals grew on board the International Space Station for 11 weeks in 2007, yielding single crystals of the wild-type L-PGDS and the C65A mutant, both of which diffracted at around 1.0 A resolution. The crystal quality was markedly improved through the use of a high viscosity precipitant solution in microgravity, in combination with the use of a highly purified protein. PMID- 21643439 TI - Fc-DIRECTED ANTIBODY CONJUGATION OF MAGNETIC NANOPARTICLES FOR ENHANCED MOLECULAR TARGETING. AB - In this study, we report the fabrication of engineered iron oxide magnetic nanoparticles (MNPs) functionalized with anti-human epidermal growth factor receptor type 2 (HER2) antibody to target the tumor antigen HER2. The Fc-directed conjugation of antibodies to the MNPs aids their efficient immunospecific targeting through free Fab portions. The directional specificity of conjugation was verified on a macrophage cell line. Immunofluorescence studies on macrophages treated with functionalized MNPs and free anti-HER2 antibody revealed that the antibody molecules bind to the MNPs predominantly through their Fc portion. Different cell lines with different HER2 expression levels were used to test the specificity of our functionalized nanoprobe for molecular targeting applications. The results of cell line targeting demonstrate that these engineered MNPs are able to differentiate between cell lines with different levels of HER2 expression. PMID- 21643440 TI - Reconstruction Using Witness Complexes. AB - We present a novel reconstruction algorithm that, given an input point set sampled from an object S, builds a one-parameter family of complexes that approximate S at different scales. At a high level, our method is very similar in spirit to Chew's surface meshing algorithm, with one notable difference though: the restricted Delaunay triangulation is replaced by the witness complex, which makes our algorithm applicable in any metric space. To prove its correctness on curves and surfaces, we highlight the relationship between the witness complex and the restricted Delaunay triangulation in 2d and in 3d. Specifically, we prove that both complexes are equal in 2d and closely related in 3d, under some mild sampling assumptions. PMID- 21643441 TI - A novel synthesis of N(1)-(substituted)-pyrimido[5,4-e]-1,2,4-triazine-5,7(1H,6H) diones. AB - A new synthesis of N(1)-(substituted)-pyrimido[5,4-e]-1,2,4-triazine-5,7(1H,6H) diones, which are analogues of the natural product toxoflavin, is reported. Condensation of preformed alkyl or aryl hydrazones with 6-chloro-3-methyl-5 nitrouracil efficiently provides pyrimidotriazinediones in a three-step process that broadens the scope of R(1) substituents. PMID- 21643442 TI - Quantification of the Multiplexing Effects in Multi-Pinhole Small Animal SPECT: A Simulation Study. AB - Our goal is to study the trade-off between image degradation and improved detection efficiency and resolution from allowing multiplexing in multi-pinhole (MPH) SPECT, and to determine the optimal pinhole number for MPH design. We used an analytical 3D MPH projector and two digitized phantoms: the mouse whole body (MOBY) phantom and a hot sphere phantom to generate noise-free and noisy projections, simulating pinhole collimators fitted with pre-studied pinhole patterns. We performed three schemes to achieve different degrees of multiplexing: 1. Fixed magnification and detection efficiency; 2. Fixed detection efficiency and changed magnification; 3. Fixed magnification and changed detection efficiency. We generated various noisy data sets by simulating Poisson noise using differently scaled noise-free projections and obtained 20 noise realizations for each setting. All datasets were reconstructed using 3D MPH ML-EM reconstruction method. We analyzed the quantitative accuracy by the normalized mean-square-error. We evaluated the image contrast for the hot sphere phantom simulation, and also the image noise by the average normalized-standard-deviation of certain pixels for different degrees of multiplexing. Generally, no apparent artifacts were observed in the reconstructed images, illustrating the effectiveness of reconstructions. Bias increased for increased degree of multiplexing. Contrast was not significantly affected by multiplexing in the specific simulation scheme (1). Scheme (2) showed that excessive multiplexing to improve image resolution would not improve the overall trade-off of bias and noise compared to no multiplexing. However, scheme (3) showed that when comparing to no multiplexing, the trade-off improved initially with increased multiplexing by allowing more number of pinholes to improve detection efficiency. The trade off reached a maximum and decreased with further multiplexing due to image degradation from increased bias. The optimal pinhole number was 7 for a compact camera with size of 12 cm * 12 cm and 9 for a standard gamma camera with size of 40 cm * 40 cm in this scheme. We conclude that the gains in improved detection efficiency and resolution by increased multiplexing are offset by increased image degradations. All the aforementioned factors must be considered in the optimum MPH collimator design for small animal SPECT imaging. PMID- 21643443 TI - Risk as Social Context: Immigration Policy and Autism in California. AB - Motivated by the dramatic increase in autism diagnoses in recent years, research into risk factors has uncovered substantial variation in autism prevalence by race/ethnicity, SES, and geography. Less studied is the connection between autism diagnosis rates and the social and political context. In this article, we link the temporal pattern of autism diagnosis for Hispanic children in California to state and federal anti-immigrant policy, particularly ballot initiative Proposition 187, limiting access to public services for undocumented immigrants and their families. Using a population-level dataset of 1992-2003 California births linked to 1992-2006 autism case records, we show that the effects of state and federal policies toward immigrants are visible in the rise and fall of autism risk over time. The common epidemiological practice of estimating risk on pooled samples is thereby shown to obscure patterns and mis-estimate effect sizes. Finally, we illustrate how spatial variation in Hispanic autism rates reflects differential vulnerability to these policies. This study reveals not only the spillover effects of immigration policy on children's health, but also the hazards of treating individual attributes like ethnicity as risk factors without regard to the social and political environments that give them salience. PMID- 21643444 TI - NETWORK EXPLORATION VIA THE ADAPTIVE LASSO AND SCAD PENALTIES. AB - Graphical models are frequently used to explore networks, such as genetic networks, among a set of variables. This is usually carried out via exploring the sparsity of the precision matrix of the variables under consideration. Penalized likelihood methods are often used in such explorations. Yet, positive definiteness constraints of precision matrices make the optimization problem challenging. We introduce non-concave penalties and the adaptive LASSO penalty to attenuate the bias problem in the network estimation. Through the local linear approximation to the non-concave penalty functions, the problem of precision matrix estimation is recast as a sequence of penalized likelihood problems with a weighted L(1) penalty and solved using the efficient algorithm of Friedman et al. (2008). Our estimation schemes are applied to two real datasets. Simulation experiments and asymptotic theory are used to justify our proposed methods. PMID- 21643445 TI - MULTIPLE TESTING VIA FDR FOR LARGE SCALE IMAGING DATA. AB - The multiple testing procedure plays an important role in detecting the presence of spatial signals for large scale imaging data. Typically, the spatial signals are sparse but clustered. This paper provides empirical evidence that for a range of commonly used control levels, the conventional FDR procedure can lack the ability to detect statistical significance, even if the p-values under the true null hypotheses are independent and uniformly distributed; more generally, ignoring the neighboring information of spatially structured data will tend to diminish the detection effectiveness of the FDR procedure. This paper first introduces a scalar quantity to characterize the extent to which the "lack of identification phenomenon" (LIP) of the FDR procedure occurs. Second, we propose a new multiple comparison procedure, called FDR(L), to accommodate the spatial information of neighboring p-values, via a local aggregation of p-values. Theoretical properties of the FDR(L) procedure are investigated under weak dependence of p-values. It is shown that the FDR(L) procedure alleviates the LIP of the FDR procedure, thus substantially facilitating the selection of more stringent control levels. Simulation evaluations indicate that the FDR(L) procedure improves the detection sensitivity of the FDR procedure with little loss in detection specificity. The computational simplicity and detection effectiveness of the FDR(L) procedure are illustrated through a real brain fMRI dataset. PMID- 21643446 TI - Provider Perspectives about Latino Patients: Determinants of Care and Implications for Treatment. AB - Primary care settings are the gateway through which the majority of Latinos access care for their physical and mental health concerns. This study explored the perspectives of primary care providers regarding their Latino patients, particularly, issues impacting their patients' access to and utilization of services. Interviews were conducted with eight primary care providers-and analyzed using consensual qualitative research methods. In addition, observations were conducted of the primary care setting to contextualize providers' perspectives. Providers indicated that care for Latinos was impacted by several domains: (a) practical/instrumental factors that influence access to care; (b) cultural and personal factors that shape patients' presentations and views about physical and mental health and treatment practices; (c) provider cultural competence; and (d) institutional factors which highlight the context of care. In addition to recommendations for research and practice, the need for interdisciplinary collaboration between psychology and medicine in reducing ethnic minority disparities was proposed. PMID- 21643447 TI - Developing and Validating a New Classroom Climate Observation Assessment Tool. AB - The climate of school classrooms, shaped by a combination of teacher practices and peer processes, is an important determinant for children's psychosocial functioning and is a primary factor affecting bullying and victimization. Given that there are relatively few theoretically-grounded and validated assessment tools designed to measure the social climate of classrooms, our research team developed an observation tool through participatory action research (PAR). This article details how the assessment tool was designed and preliminarily validated in 18 third-, fourth-, and fifth-grade classrooms in a large urban public school district. The goals of this study are to illustrate the feasibility of a PAR paradigm in measurement development, ascertain the psychometric properties of the assessment tool, and determine associations with different indices of classroom levels of relational and physical aggression. PMID- 21643448 TI - Robotic Delivery of Complex Radiation Volumes for Small Animal Research. AB - The Small Animal Radiation Research Platform (SARRP) is a novel and complete system capable of delivering multidirectional (focal), kilo-voltage radiation fields to targets in small animals under robotic control using cone-beam CT (CBCT) image guidance. The capability of the SARRP to deliver highly focused beams to multiple animal models provides new research opportunities that more realistically bridge laboratory research and clinical translation. This paper describes the design and operation of the SARRP for precise radiation delivery. Different delivery procedures are presented which enable the system to radiate through a series of points, representative of a complex shape. A particularly interesting case is shell dose irradiation, where the goal is to deliver a high dose of radiation to the shape surface, with minimal dose to the shape interior. The ability to deliver a dose shell allows mechanistic research of how a tumor interacts with its microenvironment to sustain its growth and lead to its resistance or recurrence. PMID- 21643449 TI - Lensfree on-chip holography facilitates novel microscopy applications. PMID- 21643450 TI - A Risk-based Model Predictive Control Approach to Adaptive Interventions in Behavioral Health. AB - This paper examines how control engineering and risk management techniques can be applied in the field of behavioral health through their use in the design and implementation of adaptive behavioral interventions. Adaptive interventions are gaining increasing acceptance as a means to improve prevention and treatment of chronic, relapsing disorders, such as abuse of alcohol, tobacco, and other drugs, mental illness, and obesity. A risk-based Model Predictive Control (MPC) algorithm is developed for a hypothetical intervention inspired by Fast Track, a real-life program whose long-term goal is the prevention of conduct disorders in at-risk children. The MPC-based algorithm decides on the appropriate frequency of counselor home visits, mentoring sessions, and the availability of after-school recreation activities by relying on a model that includes identifiable risks, their costs, and the cost/benefit assessment of mitigating actions. MPC is particularly suited for the problem because of its constraint-handling capabilities, and its ability to scale to interventions involving multiple tailoring variables. By systematically accounting for risks and adapting treatment components over time, an MPC approach as described in this paper can increase intervention effectiveness and adherence while reducing waste, resulting in advantages over conventional fixed treatment. A series of simulations are conducted under varying conditions to demonstrate the effectiveness of the algorithm. PMID- 21643451 TI - Estimation of Model Parameters for Steerable Needles. AB - Flexible needles with bevel tips are being developed as useful tools for minimally invasive surgery and percutaneous therapy. When such a needle is inserted into soft tissue, it bends due to the asymmetric geometry of the bevel tip. This insertion with bending is not completely repeatable. We characterize the deviations in needle tip pose (position and orientation) by performing repeated needle insertions into artificial tissue. The base of the needle is pushed at a constant speed without rotating, and the covariance of the distribution of the needle tip pose is computed from experimental data. We develop the closed-form equations to describe how the covariance varies with different model parameters. We estimate the model parameters by matching the closed-form covariance and the experimentally obtained covariance. In this work, we use a needle model modified from a previously developed model with two noise parameters. The modified needle model uses three noise parameters to better capture the stochastic behavior of the needle insertion. The modified needle model provides an improvement of the covariance error from 26.1% to 6.55%. PMID- 21643452 TI - Young Disadvantaged Men as Fathers. AB - This article reviews the existing literature on young disadvantaged fathers' involvement with children. It first outlines the predominant theoretical perspectives regarding father involvement among resident (married and cohabiting) biological fathers, resident social fathers (unrelated romantic partners of children's mothers), and nonresident biological fathers. Second, it presents a brief discussion of the ways in which fathers contribute to childrearing. Third, it describes the socioeconomic characteristics of men who enter fatherhood at a young age, highlighting that they tend to be socioeconomically disadvantaged. Fourth, it reviews the empirical research on both antecedents of father involvement and patterns of involvement across father types. Finally, it describes the limitations of existing research and provides suggestions for future research and policy. PMID- 21643453 TI - Artemisinin production in Artemisia annua: studies in planta and results of a novel delivery method for treating malaria and other neglected diseases. AB - Artemisia annua L. produces the sesquiterpene lactone, artemisinin, a potent antimalarial drug that is also effective in treating other parasitic diseases, some viral infections and various neoplasms. Artemisinin is also an allelopathic herbicide that can inhibit the growth of other plants. Unfortunately, the compound is in short supply and thus, studies on its production in the plant are of interest as are low cost methods for drug delivery. Here we review our recent studies on artemisinin production in A. annua during development of the plant as it moves from the vegetative to reproductive stage (flower budding and full flower formation), in response to sugars, and in concert with the production of the ROS, hydrogen peroxide. We also provide new data from animal experiments that measured the potential of using the dried plant directly as a therapeutic. Together these results provide a synopsis of a more global view of regulation of artemisinin biosynthesis in A. annua than previously available. We further suggest an alternative low cost method of drug delivery to treat malaria and other neglected tropical diseases. PMID- 21643454 TI - False EX1 signatures caused by sample carryover during HX MS analyses. AB - Discovery of EX1 kinetics in hydrogen exchange (HX) mass spectrometry (MS) experiments is rare. Proteins follow the EX1 kinetic regime when cooperative unfolding events simultaneously expose multiple residues to solvent such that they all become deuterated together before the region is able to refold. A number of factors can contribute to what we call "false EX1" in which it appears as though EX1 occurs in a protein when it probably does not. One of the contributors to false EX1 is peptide carryover between chromatographic runs. In this work, we explore the origins of peptide carryover in HX MS, describe how carryover causes mass spectra to indicate false EX1 kinetics and then describe an optimized washing protocol that can be used to eliminate peptide carryover. A series of solvent injections was developed and found to efficiently eliminate carryover signatures such that analysis of deuterium incorporation could be reliably followed for two proteins prone to high carryover. PMID- 21643455 TI - The Independent Effects of Phonotactic Probability and Neighborhood Density on Lexical Acquisition by Preschool Children. AB - The goal of this research was to disentangle effects of phonotactic probability, the likelihood of occurrence of a sound sequence, and neighborhood density, the number of phonologically similar words, in lexical acquisition. Two word learning experiments were conducted with 4-year-old children. Experiment 1 manipulated phonotactic probability while holding neighborhood density and referent characteristics constant. Experiment 2 manipulated neighborhood density while holding phonotactic probability and referent characteristics constant. Learning was tested at two time points (immediate vs. retention) in both a naming and referent identification task, although only data from the referent identification task were analyzed due to poor performance in the naming task. Results showed that children were more accurate learning rare sound sequences than common sound sequences and this was consistent across time points. In contrast, the effect of neighborhood density varied by time. Children were more accurate learning sparse sound sequences than dense sound sequences at the immediate test point but accuracy for dense sound sequences significantly improved by the retention test without further training. It was hypothesized that phonotactic probability and neighborhood density influenced different cognitive processes that underlie lexical acquisition. PMID- 21643456 TI - He Says, She Says: Gender and Cohabitation. AB - Cohabitation has become the modal path to marriage in the United States. However, little is known about what cohabitation means to young adults today. Drawing on data from 18 focus groups (N=138) and 54 in-depth interviews with young adults, this exploratory study investigates motivations to cohabit, and examines potential gender differences in those motivations and the meanings attached to them. We find that primary motives to cohabit include spending time together, sharing expenses, and evaluating compatibility. Strong gender differences emerge in how respondents discuss these themes and how they characterize the drawbacks of cohabitation, with men more concerned about loss of freedom and women with delays in marriage. Overall, our findings suggest that gendered cultural norms governing intimate relationships extend to cohabiting unions, and point to gender differences in the perceived role of cohabitation in union formation processes. PMID- 21643457 TI - The Prevalence of Functional Quorum-Sensing Systems in Recently Emerged Vibrio cholerae Toxigenic Strains. AB - Vibrio cholerae live in aquatic environments and cause cholera disease. Like many other bacteria, V. cholerae use quorum-sensing (QS) systems to control various cellular functions, such as pathogenesis and biofilm formation. However, some V. cholerae strains are naturally QS-defective, including defective mutations in the quorum sensing master regulator HapR. Here we examined the QS functionality of 602 V. cholerae clinical and environmental strains isolated in China from 1960 2007, by measuring QS-regulated gene expression. We found that a greater percentage of the toxigenic strains (ctxAB(+)) had functional QS as compared to the non-toxigenic strains (ctxAB(-)), and that this trend increased significantly over time. We hypothesize that QS provides adaptive value in V. cholerae pathogenic settings. PMID- 21643458 TI - Implementing an adapted version of the job seekers' workshop in a residential program for patients with substance use disorders. AB - OBJECTIVES: To evaluate the feasibility and effectiveness of adapting the Job Seekers' Workshop (JSW) to a residential setting within a Massachusetts-based substance use disorder treatment agency. METHODS: Implementation of the adapted JSW consisted of a continual sequence of three weekly sessions that focused on job interview rehearsals, practice completing job applications, and identification of job leads. Data were compiled on the employment rates of the 188 patients discharged from the residential treatment program during July - December 2006 (baseline participants, n = 95) and January - June 2007 (JSW intervention participants, n = 93). The effectiveness of the adapted JSW was evaluated through a comparison of baseline and intervention participants' employment rates at discharge from residential treatment. RESULTS: Analyses indicated a trend towards a significant increase in employment at discharge for the intervention period (40.9%) compared to baseline (29.5%), chi(2)(1, N = 188) = 2.675, p = .051. CONCLUSIONS: Further evaluation of the JSW in residential settings is necessary, but this preliminary research suggests that the intervention could begin to address the need for vocational services in residential treatment for substance use disorders. PMID- 21643460 TI - The amino terminus of cGMP-dependent protein kinase Ibeta increases the dynamics of the protein's cGMP-binding pockets. AB - The type I cGMP-dependent protein kinases play critical roles in regulating vascular tone, platelet activation and synaptic plasticity. PKG I alpha and PKG Ibeta differ in their first ~100 amino acids giving each isoform unique dimerization and autoinhibitory domains with identical cGMP-binding pockets and catalytic domains. The N-terminal leucine zipper and autoinhibitory domains have been shown to mediate isoform specific affinity for cGMP. PKG Ialpha has a >10 fold higher affinity for cGMP than PKG Ibeta, and PKG Ibeta that is missing its leucine zipper has a three-fold decreased affinity for cGMP. The exact mechanism through which the N-terminus of PKG alters cGMP-affinity is unknown. In the present study, we have used deuterium exchange mass spectrometry to study how PKG Ibeta's N-terminus affects the conformation and dynamics of its cGMP-binding pockets. We found that the N-terminus increases the rate of deuterium exchange throughout the cGMP-binding domain. Our results suggest that the N-terminus shifts the conformational dynamics of the binding pockets, leading to an "open" conformation that has an increased affinity for cGMP. PMID- 21643461 TI - Preparative separation of high-purity cordycepin from Cordyceps militaris(L.) Link by high-speed countercurrent chromatography. AB - A high-speed counter-current chromatography (HSCCC) technique in a preparative scale has been applied to separate and purify cordycepin from the extract of Cordyceps militaris(L.) Link by a one-step separation. A high efficiency of HSCCC separation was achieved on a two-phase solvent system of n-hexane-n-butanol methanol-water (23:80:30:155, v/v/v/v) by eluting the lower mobile phase at a flow rate of 2 ml/min under a revolution speed of 850 rpm. HSCCC separation of 216.2 mg crude sample (contained cordycepin at 44.7% purity after 732 cation exchange resin clean-up) yielded 64.8 mg cordycepin with purity of 98.9% and 91.7% recovery. Identification of the target compound was performed by UV, IR, MS, (1)H NMR and (13)C NMR. PMID- 21643459 TI - fTwo novel biomarkers, mesothelin and HE4, for diagnosis of ovarian carcinoma. AB - INTRODUCTION: There is a need to improve the diagnosis and prognosis of ovarian carcinoma, particularly the serous type of cancer. Mesothelin and HE4 are two novel biomarkers which are expressed in serous ovarian carcinoma and can be measured in serum and other body fluids, including urine, by using ELISA. The measurement of antibodies to these markers can provide additional useful information. AREAS COVERED: A literature search was performed to as recent as December 1 2010, using the Internet (e.g. PubMed) on articles concerning mesothelin and HE4 for the diagnosis of ovarian carcinoma. The authors also included up to date recent information from the research performed in their own laboratory. EXPERT OPINION: The combination of CA125 with HE4 facilitates the triaging of women with a pelvic mass and detects more stage I/II tumors than CA125, the present 'gold standard', when used alone. Assaying urine for HE4 or mesothelin may detect early ovarian carcinoma more often than assaying serum. Antibodies to mesothelin and HE4 are more frequent in women with ovarian carcinoma or with certain types of infertility than in controls. No presently available biomarker or multi-marker panel lends itself to screening large populations of symptomless women to make possible detection of high grade serous ovarian carcinoma at stage I or II. The authors anticipate, within the next 5 years, that a greater emphasis will be given to the fact that the different subtypes of ovarian carcinoma represent different types of disease. Each different type of disease will require a different diagnostic approach and more efforts will focus on high grade serous ovarian carcinoma for which the clinical need is the greatest. PMID- 21643462 TI - From biological and social network metaphors to coupled bio-social wireless networks. AB - Biological and social analogies have been long applied to complex systems. Inspiration has been drawn from biological solutions to solve problems in engineering products and systems, ranging from Velcro to camouflage to robotics to adaptive and learning computing methods. In this paper, we present an overview of recent advances in understanding biological systems as networks and use this understanding to design and analyse wireless communication networks. We expand on two applications, namely cognitive sensing and control and wireless epidemiology. We discuss how our work in these two applications is motivated by biological metaphors. We believe that recent advances in computing and communications coupled with advances in health and social sciences raise the possibility of studying coupled bio-social communication networks. We argue that we can better utilise the advances in our understanding of one class of networks to better our understanding of the other. PMID- 21643463 TI - Mining Boundary Effects in Areally Referenced Spatial Data Using the Bayesian Information Criterion. AB - Statistical models for areal data are primarily used for smoothing maps revealing spatial trends. Subsequent interest often resides in the formal identification of 'boundaries' on the map. Here boundaries refer to 'difference boundaries', representing significant differences between adjacent regions. Recently, Lu and Carlin (2004) discussed a Bayesian framework to carry out edge detection employing a spatial hierarchical model that is estimated using Markov chain Monte Carlo (MCMC) methods. Here we offer an alternative that avoids MCMC and is easier to implement. Our approach resembles a model comparison problem where the models correspond to different underlying edge configurations across which we wish to smooth (or not). We incorporate these edge configurations in spatially autoregressive models and demonstrate how the Bayesian Information Criteria (BIC) can be used to detect difference boundaries in the map. We illustrate our methods with a Minnesota Pneumonia amd Influenza Hospitalization dataset to elicit boundaries detected from the different models. PMID- 21643464 TI - Using Treatment Integrity Methods to Study the Implementation Process. AB - The last decade has witnessed increased interest in the implementation and dissemination of evidence-based treatments (EBTs) for youth. Nakamura et al. (this issue) detail lessons learned over the past decade from the large-scale implementation of EBTS for children in Hawaii. This commentary discusses how lessons from Hawaii's initiative can help inform the next generation of implementation research. Specifically, we focus on how treatment integrity models and methods designed to characterize core aspects of treatment delivery can be used to study the implementation process. Using the new interactive online reporting systems developed by this research group to collect treatment integrity data offers researchers a way to determine how best to implement EBTs in community based service settings with integrity and skill. PMID- 21643465 TI - Stereochemical survey of digitoxin monosaccharides: new anticancer analogues with enhanced apoptotic activity and growth inhibitory effect on human non-small cell lung cancer cell. AB - A stereochemically diverse array of monosaccharide analogues of the trisaccharide based cardiac glycoside natural product digitoxin has been synthesized using a de novo asymmetric approach. The analogues were tested for cytotoxicity against the NCI panel of 60 human cancer cell lines and in more detail against non-small cell human lung cancer cells (NCI-H460). The results were compared with digitoxin and its aglycone digitoxigenin. Three novel digitoxin monosaccharide analogues with beta-d-digitoxose, alpha-l-rhamnose, and alpha-l-amicetose sugar moieties showed excellent selectivity and activity. Further investigation revealed that digitoxin alpha-l-rhamnose and alpha-l-amicetose analogues displayed similar anti proliferation effects, but with at least 5-fold greater potency in apoptosis induction than digitoxin against NCI-H460. This study demonstrates the ability to improve the digitoxin anti-cancer activity by modification of the stereochemistry and substitution of the carbohydrate moiety of this known cardiac drug. PMID- 21643466 TI - Wealth Accumulation and Factors Accounting for Success. AB - We use detailed income, balance sheet, and cash flow statements constructed for households in a long monthly panel in an emerging market economy, and some recent contributions in economic theory, to document and better understand the factors underlying success in achieving upward mobility in the distribution of net worth. Wealth inequality is decreasing over time, and many households work their way out of poverty and lower wealth over the seven year period. The accounts establish that, mechanically, this is largely due to savings rather than incoming gifts and remittances. In turn, the growth of net worth can be decomposed household by household into the savings rate and how productively that savings is used, the return on assets (ROA). The latter plays the larger role. ROA is, in turn, positively correlated with higher education of household members, younger age of the head, and with a higher debt/asset ratio and lower initial wealth, so it seems from cross-sections that the financial system is imperfectly channeling resources to productive and poor households. Household fixed effects account for the larger part of ROA, and this success is largely persistent, undercutting the story that successful entrepreneurs are those that simply get lucky. Persistence does vary across households, and in at least one province with much change and increasing opportunities, ROA changes as households move over time to higher return occupations. But for those households with high and persistent ROA, the savings rate is higher, consistent with some micro founded macro models with imperfect credit markets. Indeed, high ROA households save by investing in their own enterprises and adopt consistent financial strategies for smoothing fluctuations. More generally growth of wealth, savings levels and/or rates are correlated with TFP and the household fixed effects that are the larger part of ROA. PMID- 21643467 TI - Adapting Dialectical Behaviour Therapy for Children: Towards a New Research Agenda for Paediatric Suicidal and Non-Suicidal Self-Injurious Behaviours. AB - BACKGROUND: Dialectical Behaviour Therapy (DBT) has been used to treat adults and adolescents with suicidal and non-suicidal self-injury. This article describes initial progress in modifying DBT for affected pre-adolescent children. METHOD: Eleven children from regular education classes participated in a 6-week pilot DBT skills training program for children. Self-report measures of children's emotional and behavioural difficulties, social skills and coping strategies were administered at pre- and post-intervention, and indicated that the children had mild to moderate symptoms of depression, anxiety and suicidal ideation at baseline. RESULTS: Subjects were able to understand and utilise DBT skills for children and believed that the skills were important and engaging. Parents also regarded skills as important, child friendly, comprehensible and beneficial. At post-treatment, children reported a significant increase in adaptive coping skills and significant decreases in depressive symptoms, suicidal ideation and problematic internalising behaviours. CONCLUSIONS: These promising preliminary results suggest that continued development of DBT for children with more severe clinical impairment is warranted. Progress on adapting child individual DBT and developing a caregiver training component in behavioural modification and validation techniques is discussed. PMID- 21643468 TI - Household and farm transitions in environmental context. AB - Recent debate in the literature on population, environment, and land use questions the applicability of theory that patterns of farm extensification and intensification correspond to the life course of farmers and to the life cycle of farm families. This paper extends the debate to the agricultural development of the United States Great Plains region, using unique data from 1875 to 1930 that link families to farms over time in 25 environmentally diverse Kansas townships. Results of multilevel statistical modeling indicate that farmer's age, household size, and household structure are simultaneously related to both the extent of farm operations and the intensity of land use, taking into account local environmental conditions and time trends as Kansas was settled and developed. These findings validate farm- and life cycle theories and offer support for intergenerational motivations for farm development that include both daughters and sons. Environmental variation in aridity was a key driver of farm structure. PMID- 21643469 TI - The sum of the distances between the leaves of a tree and the 'semi-regular' property. AB - Various topological indices have been put forward in different studies from bio chemistry to pure mathematics. Among them the Wiener index, the number of subtrees and the Randic index have received great attention from mathematicians. While studying the extremal problems regarding these indices among trees, one interesting phenomenon is that they share the same extremal tree structures. Much effort was devoted to the study of the correlations between these various indices. In this note we provide a common characteristic (the 'semi-regular' property) of these extremal structures with respect to the above mentioned indices, among trees with a given maximum degree. This observation leads to a more unified approach for characterizing these extremal structures. As an application/example, we illustrate the idea by studying the extremal trees regarding the sum of distances between all pairs of leaves of a tree, a new index, which recently appeared in phylogenetic tree reconstruction and the study of the neighborhood of trees. PMID- 21643470 TI - Lives in Motion: A Review of Former Foster Youth in the Context of their Experiences in the Child Welfare System. AB - In light of the poor prospects facing many former foster youth during the transition to adulthood, it is surprising that so little attention has been given to substitute care contexts during child welfare system involvement. A review of child welfare research finds that many former foster youth go through an alarming number of placement changes and report a high rate of placement into congregate care settings and other unplanned events. Future research should take explicit account of substitute care contexts in designing and carrying out studies examining the adult outcomes of aging out foster care populations. Interventions intended to circumvent high rates of movement in foster care should also be broadened to target multiple aspects of substitute care contexts. PMID- 21643471 TI - Exposure to Psychological Aggression at Work and Job Performance: The Mediating Role of Job Attitudes and Personal Health. AB - Despite the growing literature on workplace aggression and the importance of employee performance at work, few studies have examined the relation between workplace aggression and job performance. The purpose of this study was to investigate the relations between psychological aggression at work and two forms of job performance (task performance and contextual performance) and potential mediators of these relations. Based on Conservation of Resources theory and prior research, a model was developed and tested in which overall job attitudes (i.e., job satisfaction and organizational commitment) and overall personal health (i.e., physical and psychological health) fully mediate the relations between exposure to psychological aggression at work and both task performance and contextual performance. Data were obtained from a national probability sample of US workers (N = 2376) and the model was tested using structural equation modelling. The results supported the hypothesized model, demonstrating that exposure to psychological aggression at work negatively predicted both task performance and contextual performance, and that these relations were explained by decrements in job attitudes and health associated with exposure to psychological aggression at work. PMID- 21643472 TI - Synthesis of (-)-callicarpenal, a potent arthropod-repellent. AB - Callicarpenal (1), a natural terpenoid isolated from American beautyberry (Callicarpa americana), has shown significant repellent activities against mosquitoes, ticks and imported fire ants. Here we report our efficient synthetic approach to this natural product, and preliminary results of the mosquito biting deterrent effects of callicarpenal as well as its synthetic precursors and related C(8)-epimers. The synthetic strategy allows rapid access to various epimers and analogues of the natural product that can be used to explore its structure-activity relationship and optimize its biological properties. PMID- 21643473 TI - Elucidation of New Binding Interactions with the Tumor Susceptibility Gene 101 (Tsg101) Protein Using Modified HIV-1 Gag-p6 Derived Peptide Ligands. AB - Targeting protein-protein interactions is gaining greater recognition as an attractive approach to therapeutic development. An example of this may be found with the human cellular protein encoded by the tumor susceptibility gene 101 (Tsg101), where interaction with the p6 C-terminal domain of the nascent viral Gag protein is required for HIV-1 particle budding and release. This association of Gag with Tsg101 is highly dependent on a "Pro-Thr-Ala-Pro" ("PTAP") peptide sequence within the p6 protein. Although p6-derived peptides offer potential starting points for developing Tsg101-binding inhibitors, the affinities of canonical peptides are outside the useful range (K(d) values greater than 50 MUM). Reported herein are crystal structures of Tsg101 in complex with two structurally-modified PTAP-derived peptides. This data define new regions of ligand interaction not previously identified with canonical peptide sequences. This information could be highly useful in the design of Tsg101-binding antagonists. PMID- 21643474 TI - Real-time assessment and neuromuscular training feedback techniques to prevent ACL injury in female athletes. AB - Some athletes may be more susceptible to at-risk knee positions during sports activities, but the underlying causes are not clearly defined. This manuscripts synthesizes in vivo, in vitro and in-silica (computer simulated) data to delineate likely risk factors to the mechanism(s) of non-contact ACL injuries. From these identified risk factors, we will discuss newly developed real-time screening techniques that can be used in training sessions to identify modifiable risk factors. Techniques provided will target and correct altered mechanics which may reduce or eliminate risk factors and aid in the prevention of non-contact ACL injuries in high risk athletes. PMID- 21643475 TI - Why are Korean tense stops acquired so early: The role of acoustic properties. AB - Transcription-based studies have shown that tense stops appear before aspirated or lax stops in most Korean-acquiring children's speech. This order of mastery is predicted by the short lag Voice Onset Time (VOT) values of Korean tense stops, as this is the earliest acquired phonation type across languages. However, the tense stop also has greater motor demands than the other two phonation types, given its pressed voice quality (negative H1-H2) and its relatively high f0 value at vowel onset, word-initially. In order to explain the observed order of mastery of Korean stops, we need a more sensitive quantitative model of the role of multiple acoustic parameters in production and perception. This study explores the relationship between native speakers' transcriptions/categorizations of children's stop productions and three acoustic characteristics (VOT, H1-H2 and f0). The results showed that the primary acoustic parameter that adult listeners used to differentiate tense vs. non-tense stops was VOT. Listeners used VOT and the additional acoustic parameter of f0 to differentiate lax vs. aspirated stops. Thus, the early acquisition of tense stops is explained both by their short-lag VOT values and the fact that children need to learn to control only a single acoustic parameter to produce them. PMID- 21643476 TI - Variance computations for functional of absolute risk estimates. AB - We present a simple influence function based approach to compute the variances of estimates of absolute risk and functions of absolute risk. We apply this approach to criteria that assess the impact of changes in the risk factor distribution on absolute risk for an individual and at the population level. As an illustration we use an absolute risk prediction model for breast cancer that includes modifiable risk factors in addition to standard breast cancer risk factors. Influence function based variance estimates for absolute risk and the criteria are compared to bootstrap variance estimates. PMID- 21643477 TI - Solving Word Problems using Schemas: A Review of the Literature. AB - Solving word problems is a difficult task for students at-risk for or with learning disabilities (LD). One instructional approach that has emerged as a valid method for helping students at-risk for or with LD to become more proficient at word-problem solving is using schemas. A schema is a framework for solving a problem. With a schema, students are taught to recognize problems as falling within word-problem types and to apply a problem solution method that matches that problem type. This review highlights two schema approaches for 2(nd) and 3(rd)-grade students at-risk for or with LD: schema-based instruction and schema-broadening instruction. A total of 12 schema studies were reviewed and synthesized. Both types of schema approaches enhanced the word-problem skill of students at-risk for or with LD. Based on the review, suggestions are provided for incorporating word-problem instruction using schemas. PMID- 21643478 TI - Best Invariant and Minimax Estimation of Quantiles in Finite Populations. AB - The theoretical literature on quantile and distribution function estimation in infinite populations is very rich, and invariance plays an important role in these studies. This is not the case for the commonly occurring problem of estimation of quantiles in finite populations. The latter is more complicated and interesting because an optimal strategy consists not only of an estimator, but also of a sampling design, and the estimator may depend on the design and on the labels of sampled individuals, whereas in iid sampling, design issues and labels do not exist.We study estimation of finite population quantiles, with emphasis on estimators that are invariant under the group of monotone transformations of the data, and suitable invariant loss functions. Invariance under the finite group of permutation of the sample is also considered. We discuss nonrandomized and randomized estimators, best invariant and minimax estimators, and sampling strategies relative to different classes. Invariant loss functions and estimators in finite population sampling have a nonparametric flavor, and various natural combinatorial questions and tools arise as a result. PMID- 21643479 TI - A Meta-Analysis of the Convergent Validity of Self-Control Measures. AB - There is extraordinary diversity in how the construct of self-control is operationalized in research studies. We meta-analytically examined evidence of convergent validity among executive function, delay of gratification, and self- and informant-report questionnaire measures of self-control. Overall, measures demonstrated moderate convergence (r(random) = .27 [95% CI = .24, .30]; r(fixed) = .34 [.33, .35], k = 282 samples, N = 33,564 participants), although there was substantial heterogeneity in the observed correlations. Correlations within and across types of self-control measures were strongest for informant-report questionnaires and weakest for executive function tasks. Questionnaires assessing sensation seeking impulses could be distinguished from questionnaires assessing processes of impulse regulation. We conclude that self-control is a coherent but multidimensional construct best assessed using multiple methods. PMID- 21643480 TI - All-Optical Modulation of Localized Surface Plasmon Coupling in a Hybrid System Composed of Photo-Switchable Gratings and Au Nanodisk Arrays. AB - We conduct a real-time study of all-optical modulation of localized surface plasmon resonance (LSPR) coupling in a hybrid system that integrates a photo switchable optical grating with a gold nanodisk array. This hybrid system enables us to investigate two important interactions: 1) LSPR-enhanced grating diffraction, and 2) diffraction-mediated LSPR in the Au nanodisk array. The physical mechanism underlying these interactions was analyzed and experimentally confirmed. With its advantages in cost-effective fabrication, easy integration, and all-optical control, the hybrid system described in this work could be valuable in many nanophotonic applications. PMID- 21643481 TI - The ecology of tick-transmitted infections in the redwood chipmunk (Tamias ochrogenys). AB - The redwood chipmunk contributes to the maintenance of tick-borne diseases in northern California. The range of redwood chipmunks overlaps that of western black-legged ticks and tick-borne disease, including granulocytic anaplasmosis and Lyme borreliosis. Chipmunks have high Anaplasma phagocytophilum PCR- and seroprevalence, are infested with a diversity of Ixodes spp. ticks, and are reservoir competent for Borrelia burgdorferi. We hypothesized that chipmunks could maintain tick-borne disease on the forest floor while also potentially bridging infection to arboreal sciurids as well. We used radio-telemetry to evaluate chipmunk movement and use of trees, characterized burrows, described prevalence of tick-borne disease, and identified ticks on these chipmunks. A total of 192 chipmunks from Hendy Woods, Mendocino County, California, USA, was evaluated between November 2005 and April 2009. The mean density was 2.26-5.8 chipmunks/ha. The longest detected life span was 3 years. Female weights ranged from 80-120 g and males from 80-180 g. The A. phagocytophilum and Borrelia spp. seroprevalence was 21.4% and 24.7%, respectively, and PCR prevalence for these pathogens was 10.6% and 0%, respectively. Ixodes spp. ticks included I. angustus, I. ochotonae, I. pacificus, and I. spinipalpis. The mean infestation level was 0.92 ticks/chipmunk. Based on telemetry of 11 chipmunks, the greatest distance traveled ranged from 0.14-0.63 km for females and 0.1-1.26 km for males. Areas occupied by chipmunks ranged from 0.005-0.24 km(2) for females and 0.006-0.73 km(2) for males. On 3 occasions, chipmunks were found in trees. Burrows were identified under a moss-covered redwood log, deep under a live redwood tree, under a Douglas fir log, in a clump of huckleberry, in a root collection from an overturned Douglas fir tree, and in a cluster of exposed huckleberry roots. The biology of the redwood chipmunk has multiple features that allow it to be an important reservoir host for tick-borne disease in northwestern California. PMID- 21643482 TI - Basic Information Processing Abilities at 11 years Account for Deficits in IQ Associated with Preterm Birth. AB - Although it is well established that preterms as a group do poorly relative to their full-term peers on tests of global cognitive functioning, the basis for this relative deficiency is less understood. The present paper examines preterm deficits in core cognitive abilities and determines their role in mediating preterm/full-term differences in IQ. The performance of 11-year-old children born preterm (birth weight <1750g) and their full-term controls were compared on a large battery of 15 tasks, covering four basic cognitive domains -- memory, attention, speed of processing and representational competence. The validity of these four domains was established using latent variables and confirmatory factor analysis (CFA). Preterms showed pervasive deficits within and across domains. Additionally, preterm deficits in IQ were completely mediated by these four cognitive domains in a structural equation model involving a cascade from elementary abilities (attention and speed), to more complex abilities (memory and representational competence), to IQ. The similarity of findings to those obtained with this cohort in infancy and toddlerhood suggest that preterm deficits persist - across time, across task, and from the non-verbal to the verbal period. PMID- 21643483 TI - A geometric characterization of optimal designs for regression models with correlated observations. AB - We consider the problem of optimal design of experiments for random effects models, especially population models, where a small number of correlated observations can be taken on each individual, while the observations corresponding to different individuals are assumed to be uncorrelated. We focus on c-optimal design problems and show that the classical equivalence theorem and the famous geometric characterization of Elfving (1952) from the case of uncorrelated data can be adapted to the problem of selecting optimal sets of observations for the n individual patients. The theory is demonstrated by finding optimal designs for a linear model with correlated observations and a nonlinear random effects population model, which is commonly used in pharmacokinetics. PMID- 21643484 TI - Eyes on the Block: Measuring Urban Physical Disorder Through In-Person Observation. AB - In this paper, we present results from measuring physical disorder in Los Angeles neighborhoods. Disorder measures came from structured observations conducted by trained field interviewers. We examine inter-rater reliability of disorder measures in depth. We assess the effects of observation conditions on the reliability of reporting. Finally, we examine the relationships between disorder, other indicators of neighborhood status, and selected individual outcomes.Our results indicate that there is considerable variation in the level of agreement among independent observations across items, although overall agreement is moderate to high. Durable indicators of disorder provide the most reliable measures of neighborhood conditions. Circumstances of observation have statistically significant effects on the observers' perceived level of disorder. Physical disorder is significantly related to other indicators of neighborhood status, and to children's reading and behavior development. This result suggests a need for further research into the effects of neighborhood disorder on children. PMID- 21643485 TI - Effects of chronic, heavy cannabis use on executive functions. AB - This case describes the clinical course of a cannabis-dependent individual entering a 12-week abstinence-based research program. The case illustrates the effects of chronic, heavy cannabis use on executive functions at three time points: 1) 24 hours of abstinence; 2) 4 weeks of abstinence; and 3) 12 weeks of abstinence. It is followed by discussions by two clinical psychologists and a psychiatrist. The findings described here have important clinical implications, as executive functions have a vital role in treatment participation and in sustaining recovery. It should be of particular interest to clinicians who work with people with cannabis use disorders. PMID- 21643486 TI - Mexican-American Males Providing Personal Care for their Mothers. AB - We know little about Mexican-American (MA) family adaptation to critical events in the informal caregiving experience but, in these days of economic and social turmoil, sons must sometimes step up to provide personal care for their aging mothers. This article compares two empirically real cases of MA males who provided such care, in lieu of a female relative. The cases are selected from a federally-funded, descriptive, longitudinal, mixed methods study of 110 MA caregivers and their care recipients. In case-oriented research, investigators can generate propositions (connected sets of statements) that reflect their findings and conclusions, and can be tested against subsequent cases: Caregiving strain and burden in MA males may have more to do with physical and emotional costs than financial ones; MA males providing personal care for their mothers adopt a matter-of-fact approach as they act "against taboo"; and this approach is a new way to fulfill family obligations. PMID- 21643488 TI - Empirical Behavioral Models to Support Alternative Tools for the Analysis of Mixed-Priority Pedestrian-Vehicle Interaction in a Highway Capacity Context. AB - This paper presents behavioral-based models for describing pedestrian gap acceptance at unsignalized crosswalks in a mixed-priority environment, where some drivers yield and some pedestrians cross in gaps. Logistic regression models are developed to predict the probability of pedestrian crossings as a function of vehicle dynamics, pedestrian assertiveness, and other factors. In combination with prior work on probabilistic yielding models, the results can be incorporated in a simulation environment, where they can more fully describe the interaction of these two modes. The approach is intended to supplement HCM analytical procedure for locations where significant interaction occurs between drivers and pedestrians, including modern roundabouts. PMID- 21643487 TI - Intracellular delivery of RNA-based therapeutics using aptamers. AB - The clinical potential of siRNAs for silencing genes critical to disease progression is clear, but a fail-proof method for delivering siRNAs to the cytoplasm of diseased tissues or cells has yet to be identified. A variety of delivery approaches have been explored to directly or indirectly couple siRNAs to delivery vehicles. This review explores the use of synthetic single-stranded DNA and RNA aptamers as a means to deliver siRNAs, shRNAs and antisense oligonucleotides for therapeutic intervention. Topics covered include: the advantages and challenges of using aptamers as delivery tools; current aptamer mediated siRNA delivery platforms for the treatment of cancer and HIV; and emerging methodologies for the identification of aptamers capable of internalizing into target cell types. PMID- 21643489 TI - Anisotropic Picosecond X-ray Solution Scattering from Photo-selectively Aligned Protein Molecules. AB - Anisotropic X-ray scattering patterns of transiently aligned protein molecules in solution are measured by using pump-probe X-ray solution scattering. When a linearly polarized laser pulse interacts with an ensemble of molecules, the population of excited molecules is created with their transition dipoles preferentially aligned along the laser polarization direction. We measured the X ray scattering from the myoglobin protein molecules excited by a linearly polarized, short laser pulse and obtained anisotropic scattering patterns on 100 ps time scale. An anisotropic scattering pattern contains higher structural information content than a typical isotropic pattern available from randomly oriented molecules. In addition, multiple independent diffraction patterns measured by using various laser polarization orientations will give substantially increased amount of structural information compared with a single isotropic pattern. By monitoring the temporal change of the anisotropic scattering pattern from 100 ps to 1 MUs, we observed the orientational dynamics of photo-generated myoglobin with the rotational diffusion time of ~15 ns. PMID- 21643490 TI - What's in a drop? Correlating observations and outcomes to guide macromolecular crystallization experiments. AB - Observations of crystallization experiments are classified as specific outcomes and integrated through a phase diagram to visualize solubility and thereby direct subsequent experiments. Specific examples are taken from our high-throughput crystallization laboratory which provided a broad scope of data from 20 million crystallization experiments on 12,500 different biological macromolecules. The methods and rationale are broadly and generally applicable in any crystallization laboratory. Through a combination of incomplete factorial sampling of crystallization cocktails, standard outcome classifications, visualization of outcomes as they relate chemically and application of a simple phase diagram approach we demonstrate how to logically design subsequent crystallization experiments. PMID- 21643491 TI - Computational Hemodynamics Framework for the Analysis of Cerebral Aneurysms. AB - Assessing the risk of rupture of intracranial aneurysms is important for clinicians because the natural rupture risk can be exceeded by the small but significant risk carried by current treatments. To this end numerous investigators have used image-based computational fluid dynamics models to extract patient-specific hemodynamics information, but there is no consensus on which variables or hemodynamic characteristics are the most important. This paper describes a computational framework to study and characterize the hemodynamic environment of cerebral aneurysms in order to relate it to clinical events such as growth or rupture. In particular, a number of hemodynamic quantities are proposed to describe the most salient features of these hemodynamic environments. Application to a patient population indicates that ruptured aneurysms tend to have concentrated inflows, concentrated wall shear stress distributions, high maximal wall shear stress and smaller viscous dissipation ratios than unruptured aneurysms. Furthermore, these statistical associations are largely unaffected by the choice of physiologic flow conditions. This confirms the notion that hemodynamic information derived from image-based computational models can be used to assess aneurysm rupture risk, to test hypotheses about the mechanisms responsible for aneurysm formation, progression and rupture, and to answer specific clinical questions. PMID- 21643492 TI - Mechanisms of Family Impact on African American Adolescents' HIV-Related Behavior. AB - A longitudinal model that tested mediating pathways between protective family processes and HIV-related behavior was evaluated with 195 African American youth. Three waves of data were collected when the youth were 13, 15, and 19 years old. Evidence of mediation and temporal priority were assessed for three constructs: academic engagement, evaluations of prototypical risk-taking peers, and affiliations with risk-promoting peers. Structural equation modeling indicated that protective family processes assessed during early adolescence were associated with HIV-related behavior during emerging adulthood and that academic engagement, evaluations of prototypical risk-taking peers, and affiliations with risk-promoting peers accounted for this association. Evidence of a specific pathway emerged: protective family processes -> academic engagement negative -> evaluations of prototypical risk-taking peers-> affiliations with risk-promoting peers-> HIV-related behavior. Academic engagement also was a direct predictor of HIV-related risk behavior. PMID- 21643493 TI - Role of Parent and Peer Relationships and Individual Characteristics in Middle School Children's Behavioral Outcomes in the Face of Community Violence. AB - This study examines processes linking inner-city community violence exposure to subsequent internalizing and externalizing problems. Hypothesized risk and protective factors from three ecological domains -- children's parent and peer relationships and individual characteristics -- were examined for mediating, moderating or independent roles in predicting problem behavior among 667 children over three years of middle school. Mediation was not found. However, parent and peer variables moderated the association between exposure and internalizing problems. Under high exposure, normally protective factors (e.g., attachment to parents) were less effective in mitigating exposure's effects than under low exposure; attachment to friends was more effective. Individual competence was independently associated with decreased internalizing problems. Variables from all domains, and exposure, were independently associated with externalizing problems. Protective factors (e.g., parent attachment) predicted decreased problems; risk factors (e.g., friends' delinquency) predicted increased problems. Results indicate community violence reduction as essential in averting inner-city adolescents' poor behavioral outcomes. PMID- 21643494 TI - Trends in Global Gender Inequality (Forthcoming, Social Forces). AB - This study investigates trends in gender inequality for the world as a whole. Using data encompassing a large majority of the world's population, we examine world trends over recent decades for key indicators of gender inequality in education, mortality, political representation, and economic activity. We find that gender inequality is declining in virtually all major domains, that the decline is occurring across diverse religious and cultural traditions, and that population growth is slowing the decline because populations are growing faster in countries where there is the greatest gender inequality. PMID- 21643495 TI - When are Downward Entailing contexts identified? The case of the domain-widener ever. PMID- 21643496 TI - Real-time association rate constant measurement using combination tapered fiber optic biosensor (CTFOB) dip-probes. AB - This document reports a novel method of measuring association rate constant (k(a)) for antibody-antigen interaction using evanescent wave-based combination tapered fiber-optic biosensor (CTFOB) dip-probes. The method was demonstrated by measuring association rate constant for bovine serum albumin (BSA) and anti-BSA antibody interaction. "Direct method" was used for detection; goat anti-BSA "capture" antibodies were immobilized on the probe surfaces while the antigen (BSA) was directly labeled with Alexa 488 dye. The probes were subsequently submerged in 3 nM Labeled BSA in egg albumin (1 mg/ml). The fluorescence signal recorded was proportional to BSA anti-BSA conjugates and continuous signal was acquired suing a fiber optic spectrometer (Ocean Optics, Inc.). A 476 nm diode laser was use as an excitation source. Association constant was estimated from a plot of signal as a function of time. Measured association rate constant k(a) for the binding of BSA with anti-BSA at room temperature is (8.33 +/- 0.01) *10(4)M( 1)s(-1). PMID- 21643497 TI - Highly specific detection of IL-8 protein using combination tapered fiber-optic biosensor dip-probe. AB - We are reporting detection of IL-8 in a mixed protein solution, using combination tapered fiber-optic biosensor (CTFOB) dip-probe. Sandwich immunoassay was used as the detection technique. The specificity of the sensor was established by using two types of negative control probes. It is demonstrated that with the help of these CTFOB dip-probe we could successfully detect IL-8 with high specificity in protein mixture. The lowest detected concentration of IL-8 was 150 pM. PMID- 21643498 TI - Optimising bacterial DNA extraction from faecal samples: comparison of three methods. AB - Culture independent methods are used widely in diagnostic laboratories for infectious disease Isolation of genomic DNA from clinical samples is the first and important step in the procedure. Several procedures for extracting DNA from faecal samples have been described, including different mechanical cell disruptors. To our knowledge, the use of TissueLyser as a mechanical disruptor on faecal samples before DNA extraction has not been previously described. The purpose of the study was to implement a method for preparing faecal samples for optimal DNA extraction. Thus, three different procedures for extracting DNA from human faeces were compared. This was done either by using the mechanical disrupter by Mini BeadBeater 8, or the TissueLyser both followed by DNA purification using QIAamp DNA stool MiniKit, in comparison with DNA extractions using QIAamp DNA stool MiniKit without any prior mechanical disruption, according to manufacturer's instructions. The obtained DNA from the three procedures was analysed by DGGE, and the number of bands was compared between each procedure. There was no significant difference between the numbers of bacterial bands obtained from DGGE when using a TissueLyser or Mini BeadBeater 8, so the two different mechanical cell disruptors can be used comparably when isolating bacterial DNA from faecal samples. The QIAamp DNA stool MiniKit alone resulted in a reduced number of bands compared to the two mechanical disruption methods. PMID- 21643499 TI - Prevalence of Ehrlichia muris in Wisconsin Deer Ticks Collected During the Mid 1990s. AB - Human ehrlichiosis is due to infection by tick transmitted bacteria of the genus Ehrlichia. Based on a hypothesis for the biogeography of deer tick transmitted infections, we undertook a focused search for the Eurasian E. muris in North American deer ticks. The search was stimulated by anecdotal reports of E. muris like infection in human ehrlichiosis patients from Wisconsin. We analyzed archived adult deer ticks collected in northern Wisconsin during the 1990s by specific polymerase chain reaction for evidence of infection, and sequenced amplification products to identify E. muris. About 1% of 760 adult deer ticks collected from Spooner, Wisconsin in the 1990s contained E. muris DNA. We conclude that E. muris was present in North American deer ticks a decade ago and is likely to infect this human biting vector elsewhere in the U.S. Biogeographic theory and molecular phylogenetic methods can facilitate a targeted search for potential zoonoses. PMID- 21643500 TI - Dyslipidemia associated with chronic kidney disease. AB - Cardiovascular disease is a major cause of morbidity and mortality in patients with impaired renal function. Dyslipidemia has been established as a well-known traditional risk factor for cardiovascular disease (CVD) in the general population and it is well known that patients with chronic kidney disease (CKD) exhibit significant alterations in lipoprotein metabolism. In this review, the pathogenesis and treatment of CKD-induced dyslipidemia are discussed. Studies on lipid abnormalities in predialysis, hemodialysis and peritoneal dialysis patients are analyzed. In addition, the results of the studies that tested the effects of the hypolipidemic drugs on cardiovascular morbidity and mortality in patients with CKD are reported. PMID- 21643502 TI - Caudate nucleus and insular activation during a pain suppression paradigm comparing thermal and electrical stimulation. AB - Pain modulation is an integral function of the nervous system. It is needed to adapt to chronic stimuli. To gain insights into pain suppression mechanisms, two studies concerning the suppression of the feeling of pain with different stimulation modalities (heat vs. electrical stimuli) but using the same stimulation paradigms were compared: 15 subjects each had been stimulated on both hands under the instruction to suppress the feeling of pain. Anterior insula and DLPFC activation was seen in both single modality studies and seems to be a common feature of pain suppression, as it is absent in the interaction analyses presented here. During the task to suppress the feeling of pain, there were no consistent activations stronger under thermostimulation. But during electrostimulation, there was significantly stronger activation than during thermal stimulation in the caudate nucleus bilaterally and in the contralateral posterior insula. This may be attributed to the higher sensory-discriminative content and more demand on subjective rating and suppression of the painful electrical stimulus, compared to thermostimulation. The caudate nucleus seems to play an important role not only in the motor system but also in the modulation of the pain experience. PMID- 21643503 TI - Polyradiculopathies from schwannomatosis. AB - We describe a case of schwannomatosis presenting as radicular pain and numbness in multiple radicular nerve distributions. There were multiple peripheral nerve tumors detected by magnetic resonance imaging (MRI) at the left vestibular nerve, cauda equina, right radial nerve, thoracic paraspinal nerve, and brachial plexi. Several resected tumors have features of schwannomas, including hypercellular Antoni A areas, hypocellular Antoni B areas, Verocay bodies, and hyalinized blood vessels. The specimens are also positive for immunohistochemical staining for INI1 with diffuse nuclear staining. The findings are consistent with sporadic form of schwannomatosis. This case highlights the importance of using MRI and INI1 immunohistochemistry to differentiate familial schwannomatosis, neurofibromatosis 2 (NF2)-associated schwannomatosis, and sporadic schwannomatosis. PMID- 21643501 TI - HIV and HAART-Associated Dyslipidemia. AB - Effective highly active antiretroviral therapy (HAART) for human immunodeficiency virus-1 (HIV) infection has led to marked improvement in life-expectancy for those infected with HIV. Despite reductions in the incidence of AIDS with effective treatment, patients continue to experience considerable morbidity and mortality from non-AIDS illness such as premature cardiovascular disease, liver failure and renal failure. These morbidities, particularly premature cardiovascular disease, are thought to be related to a combination of the effects of an ageing HIV-infected population coupled with long-term effects of HIV infection and antiretroviral therapy (ART). One of the principle drivers behind the well documented increase in the risk of cardiovascular disease in HIV infected patients is dyslipidemia.This review will focus on the clinical presentation of HIV and ART-associated dyslipidemia, what is known of its patho physiology, including associations with use of specific antiretroviral medications, and suggest screening and management strategies. PMID- 21643504 TI - Brain activation during autobiographical memory retrieval with special reference to default mode network. AB - Recent neuroimaging studies have suggested that brain regions activated during retrieval of autobiographical memory (ABM) overlap with the default mode network (DMN), which shows greater activation during rest than cognitively demanding tasks and is considered to be involved in self-referential processing. However, detailed overlap and segregation between ABM and DMN remain unclear. This fMRI study focuses first on revealing components of the DMN which are related to ABM and those which are unrelated to ABM, and second on extracting the neural bases which are specifically devoted to ABM. Brain activities relative to rest during three tasks matched in task difficulty assessed by reaction time were investigated by fMRI; category cued recall from ABM, category cued recall from semantic memory, and number counting task. We delineated the overlap between the regions that showed less activation during semantic memory and number counting relative to rest, which correspond to the DMN, and the areas that showed greater or less activation during ABM relative to rest. ABM-specific activation was defined as the overlap between the contrast of ABM versus rest and the contrast of ABM versus semantic memory. The fMRI results showed that greater activation as well as less activation during ABM relative to rest overlapped considerably with the DMN, indicating that the DMN is segregated to the regions which are functionally related to ABM and the regions which are unrelated to ABM. ABM specific activation was observed in the left-lateralized brain regions and most of them fell within the DMN. PMID- 21643505 TI - NRF2 Oxidative Stress Induced by Heavy Metals is Cell Type Dependent. AB - Exposure to metallic environmental toxicants has been demonstrated to induce a variety of oxidative stress responses in mammalian cells. The transcription factor Nrf2 is activated in response to oxidative stress and coordinates the expression of antioxidant gene products. In this study, we describe the development of an Nrf2-specific reporter gene assay that can be used to study the oxidative stress response in multiple cell types. Using five different cell lines, the Nrf2-activating potency of twenty metals was assessed across a range of concentrations. While ten of the metals tested (cadmium, cobalt, copper, gold, iron, lead, mercury, silver, sodium arsenite and zinc) stimulated Nrf2-dependent transcriptional activity in at least three of the engineered cell lines, only three (cadmium, copper and sodium arsenite) were active in all five cell lines. A comparison of metal-induced Nrf2 transcriptional activation revealed significant differences in the absolute magnitude of activation as well as the relative potencies between the cell lines tested. However, there was no direct correlation between activity and potency. Taken together, these results show that the capacity to stimulate Nrf2 activity and relative potencies of these test compounds are highly dependent on the cell type tested. Since oxidative stress is thought to be involved in the mode of action of many toxicological studies, this observation may inform the design of paradigms for toxicity testing for toxicant prioritization and characterization. PMID- 21643506 TI - A High Throughput Assay for Discovery of Bacterial beta-Glucuronidase Inhibitors. AB - CPT-11 is a widely-used anti-cancer drug that is converted in vivo to its active metabolite, SN-38. In the liver, enzymes detoxify SN-38 by coupling it to a glucuronidate moiety and this inactive compound (SN-38G) is excreted into the gastrointestinal tract. In the intestine, commensal bacteria convert the SN-38G back to the active and toxic SN-38 using bacterial beta-glucuronidase enzyme (GUS). This intestinal SN-38 causes debilitating diarrhea that prevents dose intensification and efficacy in a significant fraction of patients undergoing CPT 11 treatment for cancer. This CPT-11 metabolic pathway suggests that small molecule inhibitors of GUS may have utility as novel therapeutics for prevention of dose-limiting diarrhea resulting from CPT-11 therapy. To identify chemical inhibitors of GUS activity, we employed and validated a high throughput, fluorescence-based biochemical assay and used this assay to screen a compound library. Novel inhibitors of GUS were identified with IC(50) values ranging from 50 nM to 4.8 uM. These compounds may be useful as chemical probes for use in proof-of-concept experiments designed to determine the efficacy of GUS inhibitors in altering the intestinal metabolism of drugs. Our results demonstrate that this high throughput assay can be used to identify small molecule inhibitors of GUS. PMID- 21643507 TI - Two High Throughput Screen Assays for Measurement of TNF-alpha in THP-1 Cells. AB - Tumor Necrosis Factor-alpha (TNF-alpha), a secreted cytokine, plays an important role in inflammatory diseases and immune disorders, and is a potential target for drug development. The traditional assays for detecting TNF-alpha, enzyme linked immunosorbent assay (ELISA) and radioimmunoassay, are not suitable for the large size compound screens. Both assays suffer from a complicated protocol, multiple plate wash steps and/or excessive radioactive waste. A simple and quick measurement of TNF-alpha production in a cell based assay is needed for high throughput screening to identify the lead compounds from the compound library. We have developed and optimized two homogeneous TNF-alpha assays using the HTRF (homogeneous time resolved fluorescence) and AlphaLISA assay formats. We have validated the HTRF based TNF-alpha assay in a 1536-well plate format by screening a library of 1280 pharmacologically active compounds. The active compounds identified from the screen were confirmed in the AlphaLISA TNF-alpha assay using a bead-based technology. These compounds were also confirmed in a traditional ELISA assay. From this study, several beta adrenergic agonists have been identified as TNF-alpha inhibitors. We also identified several novel inhibitors of TNF-alpha, such as BTO-1, CCG-2046, ellipticine, and PD 169316. The results demonstrated that both homogeneous TNF-alpha assays are robust and suitable for high throughput screening. PMID- 21643508 TI - Reply to Abramowitz et al.: Animal Models of OCD. PMID- 21643509 TI - Full Range Complex Spectral Domain Optical Coherence Tomography for Volumetric Imaging at 47, 000 A Scans per Second. AB - In this paper, we demonstrate a high speed spectral domain optical coherence tomography (SDOCT) system capable of achieving full range complex imaging at 47 kHz line scan rate. By applying beam-offset method, a constant modulation frequency is introduced into each B-scan that enables reconstruction of the full range complex SDOCT images of in vivo tissue samples. To make use of the full capacity of detection camera used in the system, system control software is developed that streams the raw spectral fringe data directly into the computer memory. In order to assess performance of the high speed full range SDOCT system for imaging biological specimen, we present results imaged from the cuticle of fingernail of a human volunteer in vivo, and from the chicken embryos ex vivo. We also show the high sensitivity advantages of full range complex imaging as compared to the conventional SDOCT. To the best of our knowledge, 47,000 A-scan imaging rate is the highest imaging rate ever been reported for full range complex imaging. Notwithstanding, the method reported here has no limitations on the imaging speed, thus offers a useful tool to achieve volumetric imaging of living samples where the high sensitivity region around zero-delay line in the system can be utilized for imaging. PMID- 21643510 TI - Eyes on the prize: The longitudinal benefits of goal focus on progress toward a weight loss goal. AB - Past research suggests that focusing on what has not yet been accomplished (goal focus) signals a lack of progress towards one's high commitment goals and inspires greater motivation than does focusing on what has already been accomplished (accomplishment focus). The present investigation extends this research to a longitudinal, important domain by exploring the consequences of focusing on one's goals versus accomplishments when pursuing a weight loss goal. Participants were tracked over the course of a 12-week weight loss program that utilized weekly group discussions and a companion website to direct participants' focus toward their end weight loss goal or toward what they had already achieved. Goal-focused participants reported higher levels of commitment to their goal and, ultimately, lost more weight than did accomplishment-focused and no focus control participants. Accomplishment-focused participants did not differ from controls on any measure. PMID- 21643511 TI - Natural Underwater Adhesives. AB - The general topic of this review is protein-based underwater adhesives produced by aquatic organisms. The focus is on mechanisms of interfacial adhesion to native surfaces and controlled underwater solidification of natural water-borne adhesives. Four genera that exemplify the broad range of function, general mechanistic features, and unique adaptations are discussed in detail: blue mussels, acorn barnacles, sandcastle worms, and freshwater caddisfly larva. Aquatic surfaces in nature are charged and in equilibrium with their environment, populated by an electrical double layer of ions as well as adsorbed natural polyelectrolytes and microbial biofilms. Surface adsorption of underwater bioadhesives likely occurs by exchange of surface bound ligands by amino acid sidechains, driven primarily by relative affinities and effective concentrations of polymeric functional groups. Most aquatic organisms exploit modified amino acid sidechains, in particular phosphorylated serines and hydroxylated tyrosines (dopa), with high-surface affinity that form coordinative surface complexes. After delivery to the surfaces as a fluid, permanent natural adhesives solidify to bear sustained loads. Mussel plaques are assembled in a manner superficially reminiscent of in vitro layer-by-layer strategies, with sequentially delivered layers associated through Fe(dopa)(3) coordination bonds. The adhesives of sandcastle worms, caddisfly larva, and barnacles may be delivered in a form somewhat similar to in vitro complex coacervation. Marine adhesives are secreted, or excreted, into seawater that has a significantly higher pH and ionic strength than the internal environment. Empirical evidence suggests these environment triggers could provide minimalistic, fail-safe timing mechanisms to prevent premature solidification (insolubilization) of the glue within the secretory system, yet allow rapid solidification after secretion. Underwater bioadhesives are further strengthened by secondary covalent curing. PMID- 21643512 TI - Perceptual Expertise as a Shift from Strategic Interference to Automatic Holistic Processing. AB - Holistic processing was initially characterized a unique hallmark of face perception (e.g., Young et al., 1987) and later argued a domain-general marker of perceptual expertise (e.g., Gauthier et al., 1998). More recently, evidence for holistic processing - measured by interference from task-irrelevant parts - was obtained in novices, raising questions for its usefulness as a test of expertise. Indeed, recent studies use the same task to make opposite claims: Hsiao & Cottrell (2009) found more interference in novices than experts for Chinese characters, while Wong, Palmeri & Gauthier (2009) found more interference in experts than novices with objects. Offering a resolution to this paradox, our work on the perception of musical notation (Wong & Gauthier, in press) suggests that expert and novice interference effects represent two ends of a continuum: interference is initially strategic and contextual, but becomes more automatic as holistic processing develops with the acquisition of perceptual expertise. PMID- 21643513 TI - Quantifying information transmission in eukaryotic gradient sensing and chemotactic response. AB - Eukaryotic cells are able to sense shallow chemical gradients by surface receptors and migrate toward chemoattractant sources. The accuracy of this chemotactic response relies on the ability of cells to infer gradients from the heterogeneous distribution of receptors bound by diffusing chemical molecules. Ultimately, the precision of gradient sensing is limited by the fluctuations of signaling components, including the stochastic receptor occupancy and noisy intracellular processing. Viewing the system as a Markovian communication channel, we apply techniques from information theory to derive upper bounds on the amount of information that can be reliably transmitted through a chemotactic cell. Specifically, we derive an expression for the mutual information between the gradient direction and the spatial distribution of bound receptors. We also compute the mutual information between the gradient direction and the motility direction using three different models for cell motion. Our results can be used to quantify the information loss during the various stages of directional sensing in eukaryotic chemotaxis. PMID- 21643514 TI - Effects of Prenatal Poverty on Infant Health: State Earned Income Tax Credits and Birth Weight. PMID- 21643515 TI - Effects of a Web-based intervention on women's breast health behaviors. AB - Helping women make choices to reduce cancer risk and to improve breast health behaviors is important, but the best ways to reach more people with intervention assistance is not known. To test the efficacy of a Web-based intervention designed to help women make better breast health choices, we adapted our previously tested, successful breast health intervention package to be delivered on the Internet, and then we tested it in a randomized trial. We recruited women from the general public to be randomized to either an active intervention group or a delayed intervention control group. The intervention consisted of a specialized Web site providing tailored and personalized risk information to all participants, followed by offers of additional support if needed. Follow-up at 1 year post-randomization revealed significant improvements in mammography screening in intervention women compared with control women (improvement of 13 percentage points). The intervention effects were more powerful in women who increased breast health knowledge and decreased cancer worry during intervention. These data indicate that increases in mammography can be accomplished in population-based mostly insured samples by implementing this simple, low resource intensive intervention. PMID- 21643516 TI - Health Beliefs about Osteoporosis and Osteoporosis Screening in Older Women and Men. AB - OBJECTIVE: To examine older adults' beliefs about osteoporosis and osteoporosis screening to identify barriers to screening. DESIGN: Cross-sectional mailed survey. SETTING: Western Pennsylvania. METHODS: Surveys were mailed to 1830 women and men aged 60 years and older. The survey assessed sociodemographic characteristics, osteoporosis and general health-related characteristics, and beliefs about osteoporosis severity, susceptibility, screening self-efficacy, and screening response efficacy. Analyses included Wilcoxon rank-sum tests to compare belief dimension scores, and multivariable ordinal logistic regression analyses to evaluate association between osteoporosis beliefs and potential explanatory variables. RESULTS: Surveys were completed by 1268 individuals (69.3 per cent). Mean age of respondents was 73.3 years, and most were female (58.7 per cent). Individuals demonstrated greatest belief in the severity of osteoporosis and least belief in personal susceptibility (P <.001). Older individuals believed less strongly than younger individuals in osteoporosis severity (OR, 0.95 per 1 year increase in age; 95 per cent CI, 0.92-0.97) and response efficacy (OR, 0.97 per 1-year increase in age; 95 per cent CI, 0.95-0.99). Women believed more strongly than men in osteoporosis susceptibility (OR, 1.87; 95 per cent CI, 1.38 2.53) and screening self-efficacy (OR, 2.87; 95 per cent CI, 1.17-7.07). Individuals with high self-rated health status had greater belief than those with low self-rated health status in screening self-efficacy (OR, 3.59; 95 per cent CI, 1.89-6.83). CONCLUSION: Older adults demonstrate several beliefs that may be barriers to osteoporosis screening, including low belief in susceptibility to osteoporosis. These beliefs should be targeted with patient education to improve screening rates. PMID- 21643517 TI - Characterization of Tunable Radical Metal-Carbenes: Key Intermediates in Catalytic Cyclopropanation. AB - A new class of radical metal-carbene complex has been characterized as having Fischer-like orbital interactions and adjacent pi acceptor stabilization. Density Functional Theory (DFT) along with Natural Bond Orbital (NBO) analysis and Charge Decomposition Analysis (CDA) has given insight into the electronics of this catalytic intermediate in an open-shell cobalt-porphyrin, [Co(Por)], system. The complex has a single bond from the metal to the carbene and has radical character with localized spin density on the carbene carbon. In addition, the carbene carbon is found to be nucleophilic and "tunable" through the introduction of different alpha-carbon substituents. Finally, based on these findings, rational design strategies are proposed which should lead to the enhancement of catalytic activity. PMID- 21643518 TI - Ethnicity, obesity and the metabolic syndrome: implications on assessing risk and targeting intervention. AB - Pediatric obesity threatens the future health of a growing number of children worldwide. An added challenge in identifying the patients at greatest need for intervention due to their elevated risk for future disease is that pediatric obesity and the associated metabolic syndrome manifest differently among different ethnic groups. African-Americans and Hispanics are more likely to exhibit obesity and insulin resistance and are at a higher risk for developing Type 2 diabetes. Nevertheless, using current criteria, African-American adolescents are much less likely to be diagnosed with metabolic syndrome, largely owing to lower rates of dyslipidemia. Further development is needed in ethnicity inclusive means of risk identification among adolescents to accurately target treatment toward children at highest risk for future disease and to motivate adolescent patients and their families towards lifestyle improvement. Effective targeting and intensive treatment efforts may help in avoiding future sequelae of obesity among all ethnicities. PMID- 21643520 TI - New naphthoquinones and a new delta-lactone produced by endophytic fungi from Costa Rica. AB - While searching for compounds with antimalarial activity, two new naphthoquinones, delitzchianones A (1) and B (2), were separated from Delitzchia winteri, an endophytic fungus from Costa Rica. The same search also led to a new 8-acetoxy pestalopyrone (3) and the known compound, pestalopyrone (4) from another Costa Rican endophytic fungus, Phomatospora bellaminuta. The structures of the three new compounds 1, 2 and 3 were established with extensive NMR and MS analyses. All four compounds were tested for activity in a growth / no growth Dd2 assay, but only compound 4 had measurable activity with an IC(50) value of 37 MUM. PMID- 21643521 TI - Direct Effects of Ionizing Radiation on Macromolecules. AB - In the dry or frozen states, macromolecules are damaged directly by interactions with ionizing radiation. Since gamma-rays and high-energy electrons randomly ionize orbital electrons in their path, larger molecules are more likely to suffer an interaction with these radiations. In each interaction, energy is transferred to the struck molecule, resulting in irreversibly broken covalent bonds. There is an extensive literature describing these radiation modifications in both synthetic and biopolymers. Although many different properties are measured, there emerges a similar picture of the nature of radiation damage that is common to all macromolecules. The techniques used in study of one species may be used to resolve questions raised in the other class of macromolecules. PMID- 21643519 TI - The pharmacokinetics of Toll-like receptor agonists and the impact on the immune system. AB - Toll-like receptor (TLR) ligation activates both the innate and adaptive immune systems, and plays an important role in antiviral and anti-tumor immunity. Therefore, a significant amount of effort has been devoted to exploit the therapeutic potential of TLR agonists. Depending on the therapeutic purpose, either as adjuvants to vaccine, chemotherapy or standalone therapy, TLR agonists have been administered via different routes. Both preclinical and clinical studies have suggested that the route of administration has significant effects on pharmacokinetics, and that understanding these effects is critical to the success of TLR agonist drug development. This article will summarize the pharmacokinetics of TLR agonists with different administration routes, with an emphasis on clinical studies of TLR ligands in oncologic applications. PMID- 21643522 TI - Associations between parent and child pain and functioning in a pediatric chronic pain sample: A mixed methods approach. AB - This study employed a mixed-method design to test sex-specific parent-child pain associations. Subjects were 179 chronic pain patients aged 11-19 years (mean = 14.34; 72% female) presenting for treatment at a multidisciplinary, tertiary clinic. Mothers and children completed questionnaires prior to their clinic visit, including measures of children's pain, functioning and psychological characteristics. Mothers also reported on their own pain and psychological functioning. Interviews were conducted with a sub-sample of 34 mothers and children prior to the clinic visit and analyzed using a grounded theory approach. The quantitative data suggest stronger mother-daughter than mother-son pain relationships. The qualitative data suggest that girls' pain and pain-related disability is related to an overly enmeshed mother-daughter relationship and the presence of maternal models of pain, while boys' pain and disability is linked to male pain models and criticism and to maternal worry and solicitousness. Boys and girls appear to have developmentally incongruous levels of autonomy and conformity to maternal expectations. The mixed-method data suggest distinct trajectories through which mother and father involvement may be linked to chronic pain in adolescent boys and girls. PMID- 21643523 TI - Executive Function in Preschool Children: Test-Retest Reliability. AB - Research suggests that executive function (EF) may distinguish between children who are well- or ill-prepared for kindergarten, however, little is known about the test-retest reliability of measures of EF for children. We aimed to establish a battery of EF measures that are sensitive to both development and individual differences across the preschool period using Conflict and Delay subtests that had a cool (abstract) or hot (extrinsic reward) focus. Results from 151 children in three age groups (2.5, 3.5, and 4.5) suggested acceptable same-day test-retest reliability on all but Delay-Cool subtasks. These findings will inform appropriate measurement selection and development for future studies. PMID- 21643524 TI - Lysosomes, cholesterol and atherosclerosis. AB - Cholesterol-engorged macrophage foam cells are a critical component of the atherosclerotic lesion. Reducing the sterol deposits in lesions reduces clinical events. Sterol accumulations within lysosomes have proven to be particularly hard to mobilize out of foam cells. Moreover, excess sterol accumulation in lysosomes has untoward effects, including a complete disruption of lysosome function. Recently, we demonstrated that treatment of sterol-engorged macrophages in culture with triglyceride-containing particles can reverse many of the effects of cholesterol on lysosomes and dramatically reduce the sterol burden in these cells. This article describes what is known about lysosomal sterol engorgement, discusses the possible mechanisms by which triglyceride could produce its effects, and evaluates the possible positive and negative effects of reducing the lysosomal cholesterol levels in foam cells. PMID- 21643525 TI - Diffusion tensor imaging of white matter pathology in the mouse brain. AB - Diffusion tensor imaging has been increasingly used for studying white matter pathology in rodent models of neurological diseases. Here, applications of diffusion tensor imaging in detecting major and subtle white matter pathology in the mouse CNS are reviewed, followed by several technical details that may be helpful in designing studies that involve diffusion tensor imaging of rodent brain and spinal cord. PMID- 21643526 TI - A Semi-Automatic Method for Image Analysis of Edge Dynamics in Living Cells. AB - Spatial asymmetry of actin edge ruffling contributes to the process of cell polarization and directional migration, but mechanisms by which external cues control actin polymerization near cell edges remain unclear. We designed a quantitative image analysis strategy to measure the spatiotemporal distribution of actin edge ruffling. Time-lapse images of endothelial cells (ECs) expressing mRFP-actin were segmented using an active contour method. In intensity line profiles oriented normal to the cell edge, peak detection identified the angular distribution of polymerized actin within 1 um of the cell edge, which was localized to lamellipodia and edge ruffles. Edge features associated with filopodia and peripheral stress fibers were removed. Circular statistical analysis enabled detection of cell polarity, indicated by a unimodal distribution of edge ruffles. To demonstrate the approach, we detected a rapid, nondirectional increase in edge ruffling in serum-stimulated ECs and a change in constitutive ruffling orientation in quiescent, nonpolarized ECs. Error analysis using simulated test images demonstrate robustness of the method to variations in image noise levels, edge ruffle arc length, and edge intensity gradient. These quantitative measurements of edge ruffling dynamics enable investigation at the cellular length scale of the underlying molecular mechanisms regulating actin assembly and cell polarization. PMID- 21643527 TI - Desmethyl Macrolide Analogues to Address Antibiotic Resistance: Total Synthesis and Biological Evaluation of 4,8,10-Tridesmethyl Telithromycin. AB - There is an urgent need to discover new drugs to address the pressing problem of antibiotic-resistance. Macrolide antibiotics such as erythromycin (1) are safe, broad-spectrum antibiotics used in the clinic since 1954. Herein we report the synthesis and evaluation of 4,8,10-tridesmethyl telithromycin (3), a novel desmethyl analogue of the 3rd-generation drug telithromycin (2), which is a semisynthetic derivative of 1. Analogue 3 was found to possess antibiotic activity and was superior to telithromycin (2) when tested against resistant strains of S. aureus possessing an A->T mutation at position 2058 (E. coli numbering). PMID- 21643528 TI - Metal Complexes for DNA-Mediated Charge Transport. AB - In all organisms, oxidation threatens the integrity of the genome. DNA-mediated charge transport (CT) may play an important role in the generation and repair of this oxidative damage. In studies involving long-range CT from intercalating Ru and Rh complexes to 5'-GG-3' sites, we have examined the efficiency of CT as a function of distance, temperature, and the electronic coupling of metal oxidants bound to the base stack. Most striking is the shallow distance dependence and the sensitivity of DNA CT to how the metal complexes are stacked in the helix. Experiments with cyclopropylamine-modified bases have revealed that charge occupation occurs at all sites along the bridge. Using Ir complexes, we have seen that the process of DNA-mediated reduction is very similar to that of DNA mediated oxidation. Studies involving metalloproteins have, furthermore, shown that their redox activity is DNA-dependent and can be DNA-mediated. Long range DNA-mediated CT can facilitate the oxidation of DNA-bound base excision repair proteins to initiate a redox-active search for DNA lesions. DNA CT can also activate the transcription factor SoxR, triggering a cellular response to oxidative stress. Indeed, these studies show that within the cell, redox-active proteins may utilize the same chemistry as that of synthetic metal complexes in vitro, and these proteins may harness DNA-mediated CT to reduce damage to the genome and regulate cellular processes. PMID- 21643530 TI - Impact of Parent-Provided Distraction on Child Responses to an IV Insertion. AB - This study evaluates the impact of parent-provided distraction on children's responses (behavioral, physiological, parent, and self-report) during an IV insertion. Participants were 542 children, 4 to 10 years old, randomized to an experimental group that received a parent distraction coaching intervention or to routine care. Experimental group children had significantly less cortisol responsivity (p = .026). Children that received the highest level of distraction coaching had the lowest distress on behavioral, parent report, and cortisol measures. When parents provide a higher frequency and quality of distraction, children have lower distress responses on most measures. PMID- 21643531 TI - Deficiencies in the reporting of VD and t(1/2) in the FDA approved chemotherapy drug inserts. AB - Since its release in 2006, the US Food and Drug Administration (FDA) final improved format for prescription drug labeling has revamped the comprehensiveness of drug inserts, including chemotherapy drugs. The chemotherapy drug "packets", retrieved via the FDA website and other accredited drug information reporting agencies such as the Physician Drug Reference (PDR), are practically the only available unbiased summary of information. One objective is to impartially evaluate the reporting of useful pharmacokinetic parameters, in particular, Volume of Distribution (V(D)) and elimination half-life (t(1/2)), in randomly selected FDA approved chemotherapy drug inserts. The web-accessible portable document format (PDF) files for 30 randomly selected chemotherapy drugs are subjected to detailed search and the two parameters of interest are tabulated. The knowledge of the two parameters is essential in directing patient care as well as for clinical research and since the completeness of the core FDA recommendations has been found deficient, a detailed explanation of the impact of such deficiencies is provided. PMID- 21643532 TI - High-throughput multispot single-molecule spectroscopy. AB - Solution-based single-molecule spectroscopy and fluorescence correlation spectroscopy (FCS) are powerful techniques to access a variety of molecular properties such as size, brightness, conformation, and binding constants. However, this is limited to low concentrations, which results in long acquisition times in order to achieve good statistical accuracy. Data can be acquired more quickly by using parallelization. We present a new approach using a multispot excitation and detection geometry made possible by the combination of three powerful new technologies: (i) a liquid crystal spatial light modulator to produce multiple diffraction-limited excitation spots; (ii) a multipixel detector array matching the excitation pattern and (iii) a low-cost reconfigurable multichannel counting board. We demonstrate the capabilities of this technique by reporting FCS measurements of various calibrated samples as well as single molecule burst measurements. PMID- 21643533 TI - The Role of Nonformal Education in Combating the HIV Epidemic in the Philippines and Taiwan. AB - The Philippines is experiencing a low and slow but growing prevalence of HIV, with a United Nations estimate of 6,000-11,000 cases out of a population of 91 million, and a 1.5 fold increase of new cases in 2008 compared to previous years. Previous educational programs, employing non-formal educational training techniques in the southern Philippines targeted high-risk groups such as female sex workers and their establishment managers and expanded the program to target males in the community. In comparison, Taiwan has an estimated 40,000 cases of HIV/AIDS out of a population of 23 million in 2009. Taiwan has experienced a major increase in HIV infection among injecting drug users, from 77 newly reported cases in 2003 to 2,381 newly reported cases in 2007. This paper compares and contrasts the response to the epidemic in terms of non-formal educational programs targeted and tailored to specific high risk populations in each country. PMID- 21643534 TI - Cancer Stem Cells and Epithelial-to-Mesenchymal Transition (EMT)-Phenotypic Cells: Are They Cousins or Twins? AB - Cancer stem cells (CSCs) are cells within a tumor that possess the capacity to self-renew and maintain tumor-initiating capacity through differentiation into the heterogeneous lineages of cancer cells that comprise the whole tumor. These tumor-initiating cells could provide a resource for cells that cause tumor recurrence after therapy. Although the cell origin of CSCs remains to be fully elucidated, mounting evidence has demonstrated that Epithelial-to-Mesenchymal Transition (EMT), induced by different factors, is associated with tumor aggressiveness and metastasis and these cells share molecular characteristics with CSCs, and thus are often called cancer stem-like cells or tumor-initiating cells. The acquisition of an EMT phenotype is a critical process for switching early stage carcinomas into invasive malignancies, which is often associated with the loss of epithelial differentiation and gain of mesenchymal phenotype. Recent studies have demonstrated that EMT plays a critical role not only in tumor metastasis but also in tumor recurrence and that it is tightly linked with the biology of cancer stem-like cells or cancer-initiating cells. Here we will succinctly summarize the state-of-our-knowledge regarding the molecular similarities between cancer stem-like cells or CSCs and EMT-phenotypic cells that are associated with tumor aggressiveness focusing on solid tumors. PMID- 21643535 TI - Subjective Probabilities in Household Surveys. AB - Subjective probabilities are now collected on a number of large household surveys with the objective of providing data to better understand inter-temporal decision making. Comparison of subjective probabilities with actual outcomes shows that the probabilities have considerable predictive power in situations where individuals have considerable private information such as survival and retirement. In contrast the subjective probability of a stock market gain varies greatly across individuals even though no one has private information and the outcome is the same for everyone. An explanation is that there is considerable variation in accessing and processing information. Further, the subjective probability of a stock market gain is considerably lower than historical averages, providing an explanation for the relatively low frequency of stock holding. An important research objective will be to understand how individuals form their subjective probabilities. PMID- 21643536 TI - Effects of consonant-vowel transitions in speech stimuli on cortical auditory evoked potentials in adults. AB - We examined the neural activation to consonant-vowel transitions by cortical auditory evoked potentials (AEPs). The aim was to show whether cortical response patterns to speech stimuli contain components due to one of the temporal features, the voice-onset time (VOT). In seven normal-hearing adults, the cortical responses to four different monosyllabic words were opposed to the cortical responses to noise stimuli with the same temporal envelope as the speech stimuli. Significant hemispheric asymmetries were found for speech but not in noise evoked potentials. The difference signals between the AEPs to speech and corresponding noise stimuli revealed a significant negative component, which correlated with the VOT. The hemispheric asymmetries can be referred to rapid spectral changes. The correlation with the VOT indicates that the significant component in the difference signal reflects the perception of the acoustic change within the consonant-vowel transition. Thus, at the level of automatic processing, the characteristics of speech evoked potentials appear to be determined primarily by temporal aspects of the eliciting stimuli. PMID- 21643537 TI - Relevance of basilar artery study in patients with subclavian steal phenomenon. AB - A 72-year-old male presented to the emergency department with gait instability and unclear speech. Computed tomography of the brain showed old lacunar infarcts in basal ganglia. Transcranial Doppler (TCD) sonography was normal. Extracranial Duplex sonography showed indirect hemodynamic signs of bilateral subclavian artery stenosis and both vertebral arteries also showed delayed systolic flow increase. A bilateral subclavian steal phenomenon was suspected, and arm compression tests was performed. The tests promoted reverse flow in the right VA, loss of diastolic flow in the left VA and interestingly, the normal anterograde BA flow became retrograde. Although subclavian steal is likely to be an innocuous phenomenon for the majority of our patients, it is probable that the presence of a hemodynamic effect on the basilar artery may identify those who are at special risk of neurologic symptoms. So, we recommend TCD study in all patients suffering SSP to rule out the possibility of a BA steal phenomenon. PMID- 21643538 TI - CRYO-ELECTRON MICROSCOPY DATA DENOISING BASED ON THE GENERALIZED DIGITIZED TOTAL VARIATION METHOD. AB - The energy functional used in digitalized total variation method is expanded to a general form and a generalized digitized total variation (GDTV) denoising method is obtained. We further expand this method from 2-dimensional (2D) image to 3 dimensional (3D) image processing field. Cryo-electron microscopy (cryo EM) and single particle reconstruction are becoming part of standard collection of structural techniques used for studying macromolecular assemblies. Howerver, the 3D data obtained suffers greatly from noise and degradation for the low dose electron radiation. Thus, image enhancement and noise reduction are theoretically necessary to improve the data for the subsequent segmentation and/or structure skeletonization. Although there are several methods to tackle this problem, we are pleased to find that GDTV method is very efficient and can achieve better performance. Comparative experiments are carried out to verify the enhancement achieved by the GDTV method. PMID- 21643539 TI - Genomic Instability Induced By Human Papillomavirus Oncogenes. AB - Cervical cancer is one of the leading causes of cancer death in women worldwide. Human papillomavirus (HPV) infection is necessary but not sufficient for the development of cervical cancer. Genomic instability caused by HPV allows cells to acquire additional mutations required for malignant transformation. Genomic instability in the form of polyploidy has been implicated in a causal role in cervical carcinogenesis. Polyploidy not only occurs as an early event during cervical carcinogenesis but also predisposes cervical cells to aneuploidy, an important hallmark of human cancers. Cell cycle progression is regulated at several checkpoints whose defects contribute to genomic instability.The high-risk HPVs encode two oncogenes, E6 and E7, which are essential for cellular transformation in HPV-positive cells. The ability of high-risk HPV E6 and E7 protein to promote the degradation of p53 and pRb, respectively, has been suggested as a mechanism by which HPV oncogenes induce cellular transformation. E6 and E7 abrogate cell cycle checkpoints and induce genomic instability that leads to malignant conversion.Although the prophylactic HPV vaccine has recently become available, it will not be effective for immunosuppressed individuals or those who are already infected. Therefore, understanding the molecular basis for HPV-associated cancers is still clinically relevant. Studies on genomic instability will shed light on mechanisms by which HPV induces cancer and hold promise for the identification of targets for drug development. PMID- 21643540 TI - Patterns and Predictors of Support Group Use by Custodial Grandmothers and Grandchildren. AB - Despite increasing popularity of support groups for custodial grandfamilies, the few published studies to date raise questions regarding actual support group usage and disregard predictors of such use. This study examined patterns of self reported use, need, and unmet need of both grandparent and grandchild support groups. In addition, the Andersen Social Behavioral Model was used to identify predisposing, enabling, and need factors that predicted use in a national sample of 733 grandfamilies recruited by either convenience or population-based strategies. Reported use and need for support groups were lower in the population based sample, and predictors varied by sampling strategy. The findings are discussed in terms of how they inform service providers and researchers regarding factors that influence support group use by custodial grandfamilies. PMID- 21643541 TI - Pseudomonas aeruginosa porphobilinogen synthase assembly state regulators: hit discovery and initial SAR studies. AB - Porphobilinogen synthase (PBGS) catalyzes the first common step in the biosynthesis of the essential heme, chlorophyll and vitamin B(12) heme pigments. PBGS activity is regulated by assembly state, with certain oligomers exhibiting biological activity and others either partially or completely inactive, affording an innovative means of allosteric drug action. Pseudomonas aeruginosa PBGS is functionally active as an octamer, and inactive as a dimer. We have identified a series of compounds that stabilize the inactive P. aeruginosa dimer by a computational prescreen followed by native PAGE gel mobility shift analysis. From those results, we have prepared related thiadiazoles and evaluated their ability to regulate P. aeruginosa PBGS assembly state. PMID- 21643542 TI - Body composition analysis: Cellular level modeling of body component ratios. AB - During the past two decades, a major outgrowth of efforts by our research group at St. Luke's-Roosevelt Hospital is the development of body composition models that include cellular level models, models based on body component ratios, total body potassium models, multi-component models, and resting energy expenditure body composition models. This review summarizes these models with emphasis on component ratios that we believe are fundamental to understanding human body composition during growth and development and in response to disease and treatments. In-vivo measurements reveal that in healthy adults some component ratios show minimal variability and are relatively 'stable', for example total body water/fat-free mass and fat-free mass density. These ratios can be effectively applied for developing body composition methods. In contrast, other ratios, such as total body potassium/fat-free mass, are highly variable in vivo and therefore are less useful for developing body composition models. In order to understand the mechanisms governing the variability of these component ratios, we have developed eight cellular level ratio models and from them we derived simplified models that share as a major determining factor the ratio of extracellular to intracellular water ratio (E/I). The E/I value varies widely among adults. Model analysis reveals that the magnitude and variability of each body component ratio can be predicted by correlating the cellular level model with the E/I value. Our approach thus provides new insights into and improved understanding of body composition ratios in adults. PMID- 21643543 TI - UHRF1 Links the Histone code and DNA Methylation to ensure Faithful Epigenetic Memory Inheritance. AB - Epigenetics is the study of the transmission of cell memory through mitosis or meiosis that is not based on the DNA sequence. At the molecular level the epigenetic memory of a cell is embedded in DNA methylation, histone post translational modifications, RNA interference and histone isoform variation. There is a tight link between histone post-translational modifications (the histone code) and DNA methylation, as modifications of histones contribute to the establishment of DNA methylation patterns and vice versa. Interestingly, proteins have recently been identified that can simultaneously read both methylated DNA and the histone code. UHRF1 ful-fills these requirements by having unique structural domains that allow concurrent recognition of histone modifications and methylated DNA. Herein, we review our current knowledge of UHRF1 and discuss how this protein ensures the link between histone marks and DNA methylation. Understanding the molecular functions of this protein may reveal the physiological relevance of the linkage between these layers of epigenetic marks. PMID- 21643545 TI - Effects of fluorescent dye coating of metal oxide nanoparticles on DNA scission. PMID- 21643544 TI - Obstructive sleep apnea and cardiovascular disease: evidence and underlying mechanisms. AB - A body of epidemiologic and clinical evidence dating back to the early 1960s establishes the relationships between sleep apnea and cardiovascular disease (CVD). Individuals with obstructive sleep apnea, the most common type of sleep disordered breathing, are at increased risk for coronary artery disease, congestive heart failure, and stroke. Evidence that treatment of sleep apnea with continuous positive airway pressure reduces blood pressure, improves left ventricular systolic function, and diminishes platelet activation further supports linkage between obstructive sleep apnea and CVD. Notwithstanding, complex associations between these two conditions remain largely unexplained due to dearth of systematic experimental studies. Arguably, several intermediary mechanisms including sustained sympathetic activation, intrathoracic pressure changes, and oxidative stress might be involved. Other abnormalities such as dysfunctions in coagulation factors, endothelial damage, platelet activation, and increased systemic inflammation might also play a fundamental role. This review examines evidence for the associations between obstructive sleep apnea and CVD and suggested underlying anatomical and physiological mechanisms. Specific issues pertaining to definition, prevalence, diagnosis, and treatment of sleep apnea are also discussed. Consistent with rising interest in the potential role of the metabolic syndrome, this review explores the hypothesized mediating effects of each of the components of the metabolic syndrome. PMID- 21643546 TI - Measuring segregation: an activity space approach. AB - While the literature clearly acknowledges that individuals may experience different levels of segregation across their various socio-geographical spaces, most measures of segregation are intended to be used in the residential space. Using spatially aggregated data to evaluate segregation in the residential space has been the norm and thus individual's segregation experiences in other socio geographical spaces are often de-emphasized or ignored. This paper attempts to provide a more comprehensive approach in evaluating segregation beyond the residential space. The entire activity spaces of individuals are taken into account with individuals serving as the building blocks of the analysis. The measurement principle is based upon the exposure dimension of segregation. The proposed measure reflects the exposure of individuals of a referenced group in a neighborhood to the populations of other groups that are found within the activity spaces of individuals in the referenced group. Using the travel diary data collected from the tri-county area in southeast Florida and the imputed racial-ethnic data, this paper demonstrates how the proposed segregation measurement approach goes beyond just measuring population distribution patterns in the residential space and can provide a more comprehensive evaluation of segregation by considering various socio-geographical spaces. PMID- 21643547 TI - Pairing Self-Management with Palliative Care: Intervening in Life-Limiting Illness. PMID- 21643548 TI - Suicide Note Classification Using Natural Language Processing: A Content Analysis. AB - Suicide is the second leading cause of death among 25-34 year olds and the third leading cause of death among 15-25 year olds in the United States. In the Emergency Department, where suicidal patients often present, estimating the risk of repeated attempts is generally left to clinical judgment. This paper presents our second attempt to determine the role of computational algorithms in understanding a suicidal patient's thoughts, as represented by suicide notes. We focus on developing methods of natural language processing that distinguish between genuine and elicited suicide notes. We hypothesize that machine learning algorithms can categorize suicide notes as well as mental health professionals and psychiatric physician trainees do. The data used are comprised of suicide notes from 33 suicide completers and matched to 33 elicited notes from healthy control group members. Eleven mental health professionals and 31 psychiatric trainees were asked to decide if a note was genuine or elicited. Their decisions were compared to nine different machine-learning algorithms. The results indicate that trainees accurately classified notes 49% of the time, mental health professionals accurately classified notes 63% of the time, and the best machine learning algorithm accurately classified the notes 78% of the time. This is an important step in developing an evidence-based predictor of repeated suicide attempts because it shows that natural language processing can aid in distinguishing between classes of suicidal notes. PMID- 21643549 TI - Modulation of an n->pi* interaction with alpha-fluoro groups. AB - Noncovalent interactions play an essential role in biological and chemical processes. In the main chain of common protein secondary structures, the lone pair (n) of a carbonyl oxygen is delocalized into the antibonding orbital (pi*) of the subsequent carbonyl group. Herein, experimental and computational data reveal that this n->pi* interaction can be attenuated by the inductive electron withdrawal of one or two alpha-fluoro groups in the donor. The steric effect of three alpha-fluoro groups, however, overcomes the inductive withdrawal. These data evoke a means to modulate the n->pi* interaction in peptides, proteins, and other systems. PMID- 21643550 TI - Medication Management Among Medicaid Myocardial Infarction Survivors. AB - Despite guidelines to direct appropriate medical management, the quality of care following acute myocardial infarction (AMI) may be lacking. This study characterizes medication utilization by Medicaid enrollees in the year following AMI, compares it to guidelines for secondary prevention and investigates associations with rehospitalization and survival. Using DSHS administrative claims data from Washington State, Medicaid enrollees who had an AMI in 2004 were selected. Data were de-identified and details of demographics, hospitalizations, ambulatory care, and prescriptions over the following 365 days were abstracted. Utilization of guideline-directed secondary prevention strategies was measured and associations with death and recurrent hospitalization were tested. The mortality rate was 13.4% and 38.7% were rehospitalized. Mean time to first rehospitalization was 188.6 days (SD 102.3). Prescriptions for angiotensin enzyme inhibitors or receptor blockers were initially filled by 54.0%, but year-long adherence declined to 33.3%. Beta blockers, aspirin and statins followed the same trend: 65.1% to 39.5%, 37.9% to 16.7% and 58.1% to 41.9% respectively. Twenty-two percent received all medications; 8.2% were adherent. Only the initial prescription of aspirin was significantly associated with a survival benefit (HR = 0.35, p=0.003). If the results suggested by the claims data are representative of care delivered to Medicaid enrollees, rates of application of guideline directed medication are less than optimal. To improve survival and reduce re hospitalization following AMI, changes in the access and delivery of healthcare could be implemented to improve medication management, both at time of discharge and over the year following AMI. PMID- 21643551 TI - Intermuscular and subcutaneous adipose tissue distributions differ in HIV+ versus HIV-men and women. AB - BACKGROUND: Loss of subcutaneous (SAT) with sparing of visceral (VAT) adipose tissue (AT) has been documented in HIV + men and women. Intermuscular AT (IMAT) rivals VAT in independent associations with cardiovascular risk. OBJECTIVE: To determine whether the size and distribution of IMAT differs in HIV+ vs. HIV- men and/or women. DESIGN: We used whole-body MRI to measure VAT, IMAT and four SAT compartments and compared them by HIV status using whole-body skeletal muscle (SM) or total AT (TAT) as co-variates in multi-ethnic groups of healthy HIV- (n=86) and stable HIV+ (n=76) men and women. RESULTS: The sizes of AT depots (adjusting for SM) did not differ by HIV status, except for smaller gluteal SAT (lower trunk, between L(4)-L(5) to greater trochanter) in both sexes (P<0.05). The AT distribution (adjusting for TAT) was significantly different, with larger VAT (P<0.05) and smaller gluteal and limb SAT (P<0.05) in both HIV+ sexes; IMAT increased more with TAT in HIV+ vs. HIV- men (P<0.05 for slope interaction) but there were no significant differences in women. There were significant race by HIV interactions in AT distribution with more pronounced VAT differences in non Hispanic white men and larger trunk SAT in African Americans HIV+ vs. HIV-. CONCLUSION: The AT distribution differed markedly in HIV+ vs. HIV- with limb and lower body SAT representing a smaller proportion of TAT in HIV+ in both sexes and IMAT representing a larger proportion of TAT in HIV+ vs. HIV- men. PMID- 21643552 TI - Becoming Buzz Lightyear and Other Clinical Tales: Indigenizing Disney in a World of Disability. AB - Increasingly, anthropologists are investigating the place of mass media in our lives, for we live, as Ortner (1999) notes, in a 'media-saturated world.' This paper explores the role of (globalized) children's mass media - with particular emphasis on Disney - and its influence on one particular community of consumers. The community consists of African American children who face serious disabilities and chronic illnesses, as well as the families who care for them. Disney films and characters permeate the lives and imaginations of these children and parenting kin. While the compelling power of Disney can legitimately be construed as a form of global domination, an emphasis on domination and on the consumer as unwitting victim easily underestimates the agency of the audience. PMID- 21643553 TI - Sport-Related Identities and the "Toxic Jock" AB - Little attention has been paid to the multidimensional nature of athletic involvement, which includes identity formation as well as participation in sports activities. Five hundred eighty-one sport-involved undergraduate students completed a questionnaire assessing their sport-related identities, goal orientations, primary sport ratings, and conformity to masculine norms. F-tested mean comparisons and hierarchical linear regressions were used to explore the characteristics associated with two distinct sport-related identities ("athletes" and "jocks"). Jock identity was associated with an ego-oriented approach to sports (men only), whereas athlete identity was associated with a task-oriented approach (both genders). Jock identity was positively associated with conformity to masculine norms, particularly for men, whereas athlete identity was positively associated with some masculine norms (i.e., Winning) and negatively associated with others (i.e., Playboy). These findings help to identify the correlates of a "toxic Jock" identity that may signal elevated risk for health-compromising behavior. PMID- 21643554 TI - Differential DNA damage responses in p53 proficient and deficient cells: cisplatin-induced nuclear import of XPA is independent of ATR checkpoint in p53 deficient lung cancer cells. AB - Nucleotide excision repair (NER) and ataxia telangiectasia mutated (ATM)/ATR (ATM and RAD3-related) DNA damage checkpoints are among the major pathways that affect the chemotherapeutic efficiency of the anticancer drug cisplatin. Xeroderma pigmentosum group A (XPA) protein plays a crucial role in NER including both global genome repair (GG-NER) and transcription-coupled repair (TC-NER) subpathways, and has been a potential target for improving cisplatin therapeutic effects. We report here that XPA translocates from the cytosol into the nucleus after DNA damage induced by UV irradiation and cisplatin, a mimetic of UV damage, in human cells with or without p53 deficiency. However, the damage-induced response of XPA nuclear import was significantly slower in p53-deficient cells than in p53-proficient cells. We also found that while XPA is imported into the nucleus upon cisplatin or UV damage in an ATR-dependent manner in p53-proficient A549 lung cancer cells, the ATR checkpoint pathway has no effect on the XPA nuclear import in p53-deficient H1299 lung cancer cells. Similarly, the XPA nuclear translocation is not regulated by ATM checkpoint or by p38MAPK/MK2 either. Our findings suggest that NER is independent on the major DNA damage checkpoint pathways in H1299 (p53(-/-)) cells and that DNA damage responses are mechanistically different between p53-proficient and p53-deficient cells. Our results also highlight the possibility of selectively targeting XPA nuclear import as a way to sensitize cisplatin anticancer activity, but targeting ATR/ATM dependent checkpoints may not be helpful in killing p53-deficient cancer cells. PMID- 21643555 TI - EQUINE INFECTIOUS ANEMIA VIRUS INFECTION AND IMMUNITY: LESSONS FOR AIDS VACCINE DEVELOPMENT. PMID- 21643556 TI - Development of Pre-Word-Learning Skills in Infants with Cochlear Implants. AB - Families of infants who are congenitally deaf now have the option of cochlear implantation at a very young age. In order to assess the effectiveness of early cochlear implantation, however, new behavioral procedures are needed to measure speech perception and language skills during infancy. One important component of language development is word learning-a complex skill that involves learning arbitrary relations between words and their referents. A precursor to word learning is the ability to perceive and encode intersensory relations between co occurring auditory and visual events. Recent studies in infants with normal hearing have shown that intersensory redundancies, such as temporal synchrony, can facilitate the ability to learn arbitrary pairings between speech sounds and objects (Gogate & Bahrick, 1998). To investigate the early stages of learning arbitrary pairings of sounds and objects after cochlear implantation, we used the Preferential Looking Paradigm (PLP) to assess infants' ability to associate speech sounds to objects that moved in temporal synchrony with the onset and offsets of the signals. Children with normal hearing ranging in age from 6, 9, 18, and 30 months served as controls and demonstrated the ability to learn arbitrary pairings between temporally synchronous speech sounds and dynamic visual events. Infants who received their cochlear implants (CIs) at earlier ages (7-15 months of age) performed similarly to the infants with normal hearing after about 2-6 months of CI experience. In contrast, infants who received their implants at later ages (16-25 months of age) did not demonstrate learning of the associations within the context of this experiment. Possible implications of these findings are discussed. PMID- 21643557 TI - MORPHEEINS - A NEW PATHWAY FOR ALLOSTERIC DRUG DISCOVERY. AB - The morpheein model of allosteric regulation can be applied as a novel approach to the discovery of small molecule allosteric modulators of protein function. Morpheeins are homo-oligomeric proteins where, under physiological conditions, the oligomer can dissociate, the dissociated units can change conformation, and the altered conformational state can reassociate to a structurally and functionally distinct oligomer. This phenomenon serves as a basis for allostery, as a basis for conformational diseases, as a basis for drug discovery, and may be applicable to personalized medicine such as in the prediction of drug side effects. Each of these relationships has been established for the prototype morpheein, porphobilinogen synthase, where the conformational disease is a porphyria and the drug application is in antimicrobial discovery. These data are presented along with a discussion of other drug targets for which the morpheein model of allostery may apply. Such targets include HIV integrase, TNFalpha, beta tryptase, and p53. PMID- 21643558 TI - MUC1-C Oncoprotein Blocks Terminal Differentiation of Chronic Myelogenous Leukemia Cells by a ROS-Mediated Mechanism. AB - Chronic myelogenous leukemia (CML) inevitably progresses to a blast phase by mechanisms that are not well understood. The MUC1-C oncoprotein is expressed in CML blasts but not chronic phase cells. The present studies demonstrate that treatment of KU812 and K562 CML cells with a cell-penetrating MUC1-C inhibitor, designated GO-203, is associated with increases in reactive oxygen species (ROS) and depletion of glutathione. GO-203 treatment resulted in the complete downregulation of Bcr-Abl expression and induced cell cycle arrest by a ROS mediated mechanism that was blocked by the antioxidant N-acetylcysteine. Progression of CML to blast crisis has been linked to dysregulation of Wnt/beta catenin signaling and an arrest of differentiation. The present results show that inhibition of MUC1-C induces ROS-mediated suppression of beta-catenin expression and induction of a differentiated myeloid phenotype. Our studies also show that GO-203 treatment is associated with ROS-induced decreases in ATP and loss of survival by late apoptosis/necrosis. These findings demonstrate that inhibition of the MUC1-C oncoprotein in CML cells disrupts redox balance and thereby 1) downregulates expression of both Bcr-Abl and beta-catenin and 2) induces terminal myeloid differentiation by ROS-mediated mechanisms. PMID- 21643559 TI - Quality control in tRNA charging -- editing of homocysteine. AB - All living organisms conduct protein synthesis with a high degree of accuracy maintained in the transmission and flow of information from a gene to protein product. One crucial 'quality control' point in maintaining a high level of accuracy is the selectivity by which aminoacyl-tRNA synthetases furnish correctly activated amino acids, attached to tRNA species, as the building blocks for growing protein chains. When differences in binding energies of amino acids to an aminoacyl-tRNA synthetase are inadequate, editing is used as a major determinant of enzyme selectivity. Some incorrect amino acids are edited at the active site before the transfer to tRNA (pre-transfer editing), while others are edited after transfer to tRNA at a separate editing site (post-transfer editing). Access of natural non-protein amino acids, such as homocysteine, homoserine, or ornithine to the genetic code is prevented by the editing function of aminoacyl-tRNA synthetases. Disabling editing function leads to tRNA mischarging errors and incorporation of incorrect amino acids into protein, which is detrimental to cell homeostasis and inhibits growth. Continuous homocysteine editing by methionyl tRNA synthetase, resulting in the synthesis of homocysteine thiolactone, is part of the process of tRNA aminoacylation in living organisms, from bacteria to man. Excessive homocysteine thiolactone synthesis in hyperhomocysteinemia caused by genetic or nutritional deficiencies is linked to human vascular and neurological diseases. PMID- 21643560 TI - In vitro inhibition of topoisomerase IIalpha by reduced glutathione. AB - In most cells, the major intracellular redox buffer is glutathione (GSH) and its disulfide-oxidized (GSSG) form. The GSH/GSSG system maintains the intracellular redox balance and the essential thiol status of proteins by thiol disulfide exchange. Topoisomerases are thiol proteins and are a target of thiol-reactive substances. In this study, the inhibitory effect of physiological concentration of GSH and GSSG on topoisomerase IIalpha activity in vitro was investigated. GSH (0-10 mM) inhibited topoisomerase IIalpha in a concentration-dependent manner while GSSG (1-100 uM) had no significant effect. These findings suggest that the GSH/GSSG system could have a potential in vivo role in regulating topoisomerase IIalpha activity. PMID- 21643561 TI - Unexpected phosphodiesterase activity at low pH of a dinuclear copper-beta cyclodextrin complex. AB - A surprisingly low pK(a) (4.3) for a Cu(II) bridging water was found in the presented complex, Cu(2)L, resulting in 3 orders of magnitude higher phosphodiesterase activity on BNPP than Zn(2)L at typical lysosomal pH (~5.0). PMID- 21643562 TI - Toxicity tests aiming to protect Brazilian aquatic systems: current status and implications for management. AB - The current status of toxicological tests performed with Brazilian native species was evaluated through a survey of the scientific data available in the literature. The information gathered was processed and an electronic toxicology database (http://www.inct-ta.furg.br/bd_toxicologico.php) was generated. This database provides valuable information for researchers to select sensitive and tolerant aquatic species to a large variety of aquatic pollutants. Furthermore, the toxicology database allows researchers to select species representative of an ecosystem of interest. Analysis of the toxicology database showed that ecotoxicological assays have significantly improved in Brazil over the last decade, in spite of the still relatively low number of tests performed and the restricted number of native species tested. This is because most of the research is developed in a few laboratories concentrated in certain regions of Brazil, especially in Southern and Southeast regions. Considering the extremely rich biodiversity and the large variety of aquatic ecosystems in Brazil, this finding points to the urgent need for the development of ecotoxicological studies with other groups of aquatic animals, such as insects, foraminifera, cnidarians, worms, amphibians, among others. This would help to derive more realistic water quality criteria (WQC) values, which would better protect the different aquatic ecosystems in Brazil. Finally, the toxicology database generated presents solid and science based information, which can encourage and drive the Environmental Regulatory Agencies in Brazil to derive WQC based on native species. In this context, the present paper discusses the historical evolution of ecotoxicological studies in Brazil, and how they have contributed to the improvement of the Brazilian Federal and Regional regulations for environment. PMID- 21643563 TI - Domino synthesis of 1,3,5-trisubstituted hydantoins: a DFT study. AB - The mechanism of the reaction between carbodiimides and activated alpha,beta unsaturated carboxylic acids yielding fully substituted hydantoins and variable amounts of N-acyl urea by-products was studied using density functional theory calculations. Two alternative pathways featuring N-acyl ureas and imino oxazolidinones as intermediates for the formation of the hydantoin product were taken into account. The results obtained using two different computational models indicate that the overall barriers are similar for the two pathways considered. In all cases, inclusion of a second molecule of carboxylic acid was required to mediate tautomerizations and rearrangement steps. The calculations successfully reproduce the experimentally observed regioselectivity with respect to both N acyl urea and hydantoin products. PMID- 21643564 TI - Bridged nucleic acid conjugates at 6'-thiol: synthesis, hybridization properties and nuclease resistances. AB - The bridged nucleic acid (BNA) containing a thiol at the 6'-position in the bridged structure was synthesized from the disulfide-type BNA and conjugated with various functional molecules via the thioether or the disulfide linkage post synthetically and efficiently in solution phase. The disulfide-linked conjugate was cleaved under reductive conditions derived from glutathione and an oligonucleotide bearing a free thiol was released smoothly. Conjugated functional molecules had great effects on duplex stability with the DNA complement. In contrast, the molecules little influenced the stability with the RNA complement. Moreover, the oligonucleotides with functional groups at the 6'-position had as high or higher resistances against 3'-exonuclease than phosphorothioate oligonucleotide (S-oligo). PMID- 21643565 TI - Regioselective dibromination of methyl indole-3-carboxylate and application in the synthesis of 5,6-dibromoindoles. AB - Treatment of methyl indole-3-carboxylate with bromine in acetic acid gives methyl 5,6-dibromoindole-3-carboxylate regioselectively, from which the parent 5,6 dibromoindole can be accessed via a one-pot, microwave-mediated ester hydrolysis and decarboxylation. Application of these building blocks in syntheses of natural and non-natural 5,6-dibromoindole derivatives, including meridianin F and 5,6 dibromo-2'-demethylaplysinopsin, is reported. PMID- 21643566 TI - Low frequency temperature forcing of chemical oscillations. AB - The low frequency forcing of chemical oscillations by temperature is investigated experimentally in the Belousov-Zhabotinsky (BZ) reaction and in simulations of the Oregonator model with Arrhenius temperature dependence of the rate constants. Forcing with temperature leads to modulation of the chemical frequency. The number of response cycles per forcing cycle is given by the ratio of the natural frequency to the forcing frequency and phase locking is only observed in simulations when this ratio is a whole number and the forcing amplitude is small. The global temperature forcing of flow-distributed oscillations in a tubular reactor is also investigated and synchronisation is observed in the variation of band position with the external signal, reflecting the periodic modulation of chemical oscillations by temperature. PMID- 21643567 TI - Antiferromagnetic interactions in the quarter-filled organic conductor (EDO TTF)2PF6. AB - The ground state electronic structure of the high-temperature (HT) and the low temperature (LT) phases of (EDO-TTF)(2)PF(6) is investigated using the embedded cluster approach in combination with the density functional method designed to describe the strong non-dynamic electron correlation. It is found that, in the HT phase, the unpaired electron spins located on pairs of neighbouring EDO-TTF molecules are antiferromagnetically coupled along the stacking direction with the Heisenberg exchange integral J = -655 cm(-1). In the LT phase, the unpaired spins located on the cationic EDO-TTF molecules are coupled antiferromagnetically with J values strongly alternating along the stacking axis of the crystal thus rendering it diamagnetic. The parameters of the extended Hubbard model are evaluated and the conductance properties of the two phases are estimated using these parameters. It is suggested to investigate the charge and spin excitations in the two phases of (EDO-TTF)(2)PF(6) with the use of angle-resolved photoemission spectroscopy. PMID- 21643568 TI - N-heterocyclic carbene catalyzed direct carbonylation of dimethylamine. AB - N-Heterocyclic carbene (NHC) catalyzed direct carbonylation of dimethylamine leading to the formation of DMF was successfully accomplished under metal-free conditions. The catalytic efficiency was investigated and the turnover numbers can reach as high as >300. The possible mechanism was also proposed. PMID- 21643569 TI - From static to dynamic: escaping kinetic traps in hydrazone-based dynamic combinatorial libraries. AB - Thermodynamic control over kinetically-trapped mixtures of hydrazone-based macrocycles is achieved by addition of an aromatic monohydrazide to generate dynamic combinatorial libraries (DCLs) of linear and macrocyclic oligomers. PMID- 21643570 TI - Enantioselective [2+2+2] cycloaddition of ketenes and carbon disulfide catalyzed by N-heterocyclic carbenes. AB - The chiral N-heterocyclic carbene-catalyzed [2+2+2] cycloaddition of ketenes and carbon disulfide was realized to give the cycloadduct of 1,3-oxathian-6-ones in good yields with excellent enantioselectivities. PMID- 21643571 TI - Pd-mediated new synthesis of pyrroles: their evaluation as potential inhibitors of phosphodiesterase 4. AB - A sequential Pd-mediated multi-component reaction followed by Suzuki or Heck or Sonogashira coupling in a single pot has been developed for the synthesis of functionalized pyrroles as potential inhibitors of PDE4. PMID- 21643572 TI - Site-specific immobilization of gold binding polypeptide on gold nanoparticle coated graphene sheet for biosensor application. AB - The effective and strong immobilization of enzymes on solid surfaces is required for current biological applications, such as microchips, biofuel cells, and biosensors. Gold-binding polypeptide (GBP), a genetically designed peptide, possesses unique and specific interactions with a gold surface, resulting in improved enzyme stability and activity. Herein we demonstrated an immobilization method for biosensor applications through site-specific interactions between GBP fused organophosphorus hydrolase (GBP-OPH) and gold nanoparticle-coated chemically modified graphene (Au-CMG), showing enhanced sensing capability. A flow injection biosensor was fabricated by using GBP-OPH/Au-CMG to detect paraoxons, a model pesticide, showing higher sensitivity, lower detection limit and better operating stability compared that of OPH/Au-CMG. This strategy, which integrates biotic and abiotic moieties through site-specific interactions, has a great potential for use in biosensing and bioconversion process. PMID- 21643573 TI - Multiferroic CoFe2O4-Pb(Zr(0.52)Ti(0.48))O3 core-shell nanofibers and their magnetoelectric coupling. AB - Multiferroic CoFe(2)O(4)-Pb(Zr(0.52)Ti(0.48))O(3) core-shell nanofibers have been synthesized by coaxial electrospinning in combination with a sol-gel process. The core-shell configuration of nanofibers has been verified by scanning electron microscopy and transmission electron microscopy, and the spinel structure of CoFe(2)O(4) and perovskite structure of Pb(Zr(0.52)Ti(0.48))O(3) have been confirmed by X-ray diffraction and selected area electron diffraction. The multiferroic properties of core-shell nanofibers have been demonstrated by magnetic hysteresis and piezoresponse force microscopy, and their magnetoelectric coupling has been confirmed by evolution of piezoresponse under an external magnetic field, showing magnetically induced ferroelectric domain switching and changes in switching characteristics. The lateral magnetoelectric coefficient is estimated to be 2.95 * 10(4) mV/cmOe, two orders of magnitude higher than multiferroic thin films of similar composition. PMID- 21643578 TI - Molecular interactions and structure of a supramolecular arrangement of glucose oxidase and palladium nanoparticles. AB - This paper presents studies about the molecular interactions and redox processes involved in the formation of palladium nanoparticles associated to glucose oxidase (GOx-PdNPs) in a supramolecular arrangement. The synthesis occurs in two steps, the Pd reduction and the formation of the 80 nm sized supramolecular aggregates containing multiples units of GOx associated to 3.5 nm sized PdNPs. During synthesis, GOx molecules interact with Pd salt leading to metal ion and FAD reduction probably via the thiol group of the cysteine 521 residue. For the growing of PdNPs, formic acid was necessary as a co-adjuvant reducing agent. Besides the contribution for the redox processes, GOx is also necessary for the NP stability preventing the formation of precipitates resulted from uncontrolled growing of NPs Cyclic voltammetry of the GOx-PdNPs demonstrated electroactivity of the bionanocomposite immobilized on ITO (indium-tin oxide) electrode surface and also the NP is partially blocked due to strong interaction GOx and the surface of PdNPs. Vibrational spectroscopy (FTIR) showed that significant structural changes occurred in GOx after the association to PdNP. These mechanistics and structural studies can contribute for modulation of bionanocomposites properties. PMID- 21643579 TI - Electrochemical and spectroscopic studies of ethanol oxidation on Pt stepped surfaces modified by tin adatoms. AB - Ethanol oxidation on platinum stepped surfaces vicinal to the (111) pole modified by tin has been studied to determine the role of this adatom in the oxidation mechanism. Tin has been slowly deposited so that the initial stages of the deposition take place on the step, and deposition on the terrace only occurs when the step has been completely decorated. Voltammetric and chronoamperometric experiments demonstrate that tin on the step catalyzes the oxidation. The maximum enhancement is found when the step is completely decorated by tin. FTIR experiments using normal and isotopically labeled ethanol have been used to elucidate the effect of the tin adatoms in the mechanism. The obtained results indicate that the role of tin is double: (i) when the surface has sites capable of breaking the C-C bond of the molecule, that is, when the step sites are not completely covered by tin, it promotes the oxidation of CO formed from the molecular fragments to CO(2) through a bifunctional mechanism and (ii) it catalyzes the oxidation of ethanol to acetic acid. PMID- 21643580 TI - The influence of polarizability on the dielectric spectrum of the ionic liquid 1 ethyl-3-methylimidazolium triflate. AB - This work reports for the first time the computational, frequency-dependent dielectric spectrum of the polarizable molecular ionic liquid 1-ethyl-3 methylimidazolium triflate as well as its experimental analogue. In the frequency range from 500 MHz up to 20 GHz the agreement between the computational and the experimental spectrum is quantitative. For higher frequencies up to 10 THz the agreement is still remarkably good. The experimental asymptotic limit epsilon(infinity) is 2.3. The difference in the computational value of 1.9 comes solely from the neglect of polarizability of the hydrogen atoms. For reasons of efficiency the simulations are based on the Lagrangian algorithm for the Drude oscillator model which cannot handle polarizable hydrogens. In the computational analysis the complete spectrum of the generalized dielectric constant ?(0)*(nu) is splitted into its translational and non-translational components, called dielectric conductivity theta(0)(nu) and dielectric permittivity epsilon(nu). For 1-ethyl-3-methylimidazolium triflate both components contribute with equal weight and overlap in the complete frequency range. The inclusion of polarization forces, however, is quite different for the two components: the collective non translational dynamics is accelerated and hence the dielectric permittivity is shifted to higher frequencies. The low frequency region of the dielectric conductivity is also affected while its high frequency part remains almost unchanged. Inductive effects are not only visible at high frequencies but also contribute in the sub-GHz region. The computational peak found in this region correlates with the experimental OKE-spectrum. It may be interpreted as the correlation between the induced dipole moment of the cations and the local electric field exerted by the anionic cage. PMID- 21643581 TI - Effect of alkyl chain length and hydroxyl group functionalization on the surface properties of imidazolium ionic liquids. AB - Properties of the surface of ionic liquids, such as surface tension, ordering, and charge and density profiles, were studied using molecular simulation. Two types of modification in the molecular structure of imidazolium cations were studied: the length of the alkyl side chain and the presence of a polar hydroxyl group at the end of the side chain. Four ionic liquids were considered: 1-ethyl-3 methylimidazolium tetrafluoroborate, [C(2)C(1)im][BF(4)]; 1-(2-hydroxyethyl)-3 methylimidazolium tetrafluoroborate, [C(2)OHC(1)im][BF(4)]; 1-octyl-3 methylimidazolium tetrafluoroborate, [C(8)C(1)im][BF(4)] and 1-(8-hydroxyoctyl)-3 methylimidazolium tetrafluoroborate, [C(8)OHC(1)im][BF(4)]. The surface tension was calculated using both mechanical and thermodynamic definitions, with consistent treatment of the long-range corrections. The simulations reproduce the available experimental values of surface tension with a maximum deviation of +/ 10%. This energetic characterization of the interface is completed by microscopic structural analysis of orientational ordering at the interface and density profiles along the direction normal to the interface. The presence of the hydroxyl group modifies the local structure at the interface, leading to a less organized liquid phase. The results allow us to relate the surface tension to the structural ordering at the liquid-vacuum interface. PMID- 21643582 TI - Theoretical characterization of laser- and sympathetically-cooled ions in surface electrode ion traps. AB - Using molecular dynamics simulations we characterize theoretically Coulomb clusters of laser- and sympathetically-cooled ions in a five-wire surface electrode ion trap. We show that the asymmetry of the trapping potential leads to significantly different cluster structures and ion energy distributions in comparison to conventionally used linear Paul traps and to an asymmetric segregation of the ions in bi-component Coulomb clusters. We explore the impact of our results on the implementation of sympathetic cooling of molecular ions in surface-electrode traps and discuss possible challenges for the realization of such experiments. PMID- 21643584 TI - A correlation-based predictor for pair-association in ionic liquids. AB - Pair association in Ionic Liquids is an important quantity that affects many of their physical and chemical properties. However, the association constant is a complex function of the component ions as well as of the solvent environment, and no single theory can compute or predict it with quantitative accuracy. In this work we analyze infinite-dilution association data from a number of recent conductance measurements, and develop a linear model correlating the association constant with two relevant interaction energies, i.e., (1) the dielectrically screened Coulomb attraction and hydrogen bonding between ion-pairs, and (2) the ion solvation energy, which in turn takes into account solvent-specific interactions like hydrogen-bond acidity/basicity and hydrophobic/hydrophilic interactions. The results reveal the unique nature of water as a solvent in that it affects ionic association in ways qualitatively different from other common solvents. PMID- 21643583 TI - Coarse-grained force field: general folding theory. AB - We review the coarse-grained UNited RESidue (UNRES) force field for the simulations of protein structure and dynamics, which is being developed in our laboratory over the last several years. UNRES is a physics-based force field, the prototype of which is defined as a potential of mean force of polypeptide chains in water, where all the degrees of freedom except the coordinates of alpha-carbon atoms and side-chain centers have been integrated out. We describe the initial implementation of UNRES to protein-structure prediction formulated as a search for the global minimum of the potential-energy function and its subsequent molecular dynamics and extensions of molecular-dynamics implementation, which enabled us to study protein-folding pathways and thermodynamics, as well as to reformulate the protein-structure prediction problem as a search for the conformational ensemble with the lowest free energy at temperatures below the folding-transition temperature. Applications of UNRES to study biological problems are also described. PMID- 21643585 TI - Silver nanoparticle studded porous polyethylene scaffolds: bacteria struggle to grow on them while mammalian cells thrive. AB - Silver nanoparticle studded scaffolds were prepared by exploiting the Ag(+) ion reducing activity of sophorolipids--a class of 'glycolipids' that cap the ensuing nanoparticles as well. To achieve this, the porous polyethylene scaffolds are subjected to N(2) + H(2) plasma treatment, in the first step. Subsequently the sophorolipids are covalently attached to the amine groups on the polymer surface through simple amide chemistry to yield sophorolipid grafted polymer scaffolds. These are then exposed to Ag(+) ions under appropriate conditions leading to the formation of silver nanoparticles immobilized on the polymer scaffolds. It has been found that while bacteria do not survive on these silver studded scaffolds, CHO-K1 cells thrive on them making them good candidates for tissue engineering and bio-implant applications. PMID- 21643586 TI - Spatially resolved photoconductivity of thin films formed by colloidal octapod shaped CdSe/CdS nanocrystals. AB - We studied the optical absorption and photoconductive properties of thin films consisting of core-shell octapod-shaped nanocrystals, which consisted of CdS pods that branch out from a CdSe core. The current-voltage characteristics were measured at room and cryogenic temperatures and agreed well with a phenomenological exponential fitting model, from which we could extract the sheet resistance and the average voltage barrier for the charge tunneling between the octapods. The temperature dependence of the photocurrent showed temperature activated behavior above 220 K and a non-Arrhenius exponential (T/T(0))(n) dispersion below 220 K. Furthermore, we mapped the photocurrent generation within the octapod film via scanning photocurrent microscopy, which revealed photocurrent enhancement near micron-size voids and spatial shifts of the photocurrent maxima with bias voltage. PMID- 21643587 TI - Synthesis and characterization of bis-cyclopropanated 1,3,5-tricarbonyl compounds. A combined synthetic, spectroscopic and theoretical study. AB - Bis-cyclopropanated 1,3,5-tricarbonyl compounds were prepared by a sequence of Claisen condensations and cyclopropanations. The optimization of the conditions proved to be very important to suppress retro-Claisen reactions. The conformation of these molecules was studied by experimental and computational methods. The syn/syn;syn/syn conformation is present for all derivatives. It is exclusively present in the case of the derivative containing a phenyl group located at the terminal carbon atom. In most cases, equilibria with other conformers are found. PMID- 21643588 TI - Enantioselective binding of a lanthanide(III) complex to human serum albumin studied by 1H STD NMR techniques. AB - The enantioselective binding of the (SSS)-Delta isomer of an yttrium(III) tetraazatriphenylene complex to 'drug-site II' of human serum albumin (HSA) was detected by the intensity differences of its STD (1)H NMR spectrum relative to the (RRR)-Lambda isomer, by the effect of the competitive binder to that site, N dansyl sarcosine, upon the STD spectrum of each isomer, in the presence of HSA and by 3D docking simulations. PMID- 21643589 TI - Stereoselective aza-Darzens reactions of tert-butanesulfinimines: convenient access to chiral aziridines. AB - Stereoselective synthesis of 2,3-di- and 2,2',3-tri-substituted aziridines in good yields and excellent diastereoselectivities are achieved through aza-Darzens reactions of a range of tert-butanesulfinyl aldimines and ketimines with ethyl bromoacetate. PMID- 21643590 TI - FeCl3-mediated synthesis of polysubstituted tetrahydroquinolines via domino Mannich/Friedel-Crafts reactions of aldehydes and amines. AB - A useful method to construct highly substituted tetrahydroquinolines has been developed through an iron(III) chloride-mediated domino Mannich and intramolecular Friedel-Crafts alkylation followed by intermolecular Friedel Crafts alkylation reactions of aliphatic aldehydes with aromatic amines. PMID- 21643591 TI - Nitrosyl iron complexes--synthesis, structure and biology. AB - Nitrosyl complexes of iron are formed in living species in the presence of nitric oxide. They are considered a form in which NO can be stored and stabilized within a living cell. Upon entering a topic in bioinorganic chemistry the researcher faces a wide spectrum of issues concerning synthetic methods, the structure and chemical properties of the complex on the one hand, and its biological implications on the other. The aim of this review is to present the newest knowledge on nitrosyl iron complexes, summarizing the issues that are important for understanding the nature of nitrosyl iron complexes, their possible interactions, behavior in vitro and in vivo, handling of the preparations etc. in response to the growing interest in these compounds. Herein we focus mostly on the dinitrosyl iron complexes (DNICs) due to their prevailing occurrence in NO treated biological samples. This article reviews recent knowledge on the structure, chemical properties and biological action of DNICs and some mononitrosyls of heme proteins. Synthetic methods are also briefly reviewed. PMID- 21643592 TI - Unusual carbamate-directed CH-activation at an annulated ferrocenophane framework. AB - The tetrahydroazepine-annulated [3]ferrocenophane carbamate (4) was synthesized by two different linear routes starting from the readily available alpha dimethylamino[3]ferrocenophane-ortho-carbaldehyde rac-6. The carbamate directed lithiation of 4 resulted in a selective attack at a (Cp)C-H bond at the higher substituted "lower" [3]ferrocenophane Cp-ring to eventually yield the respective ester (18) after treatment with ClCO(2)Me. PMID- 21643593 TI - Fluorescence signal transduction mechanism for immunoassay based on zinc ion release from ZnS nanocrystals. AB - In this work, a fluorescence signal transduction mechanism based on cation release from ZnS nanocrystals was developed for sandwich immunoassay. In this mechanism, ZnS nanocrystals as labels in immunoassay are dissolved by acid to release zinc ions. After pH adjustment of the dissolving solution using a basic solution, zinc-ion sensitive fluorescence indicator Fluozin-3 is added to bind with the released zinc ions for sensitive fluorescence measurement. Using mouse IgG as a model analyte, the immunoassay adopting this signal transduction mechanism demonstrates a low detection limit around 1 pM and a detection range with two orders of magnitude (1 pM to 0.5 nM). PMID- 21643594 TI - Detecting specific saccharides via a single indicator. AB - An improved synthesis of a rhodamine boronic acid indicator is reported. This compound is used in an optimized data collection protocol for wavelength- and time-dependent selectivity of sugars such as fructose and ribose derivatives. One indicator is thus used to selectively distinguish structurally related sugar analytes. PMID- 21643595 TI - Crystalline ZnO thin film by hydrothermal growth. AB - In this work, we have successfully synthesised ZnO crystal thin film with a high quality from hydrothermal reaction on sapphire substrate. The growth mechanism is clarified based on an extensive XTEM study. In addition, electrical and optical properties of the crystal thin film, which can be controlled by impurities, are characterized. The synthesis technique provides fairly high quality and cost effective substrates for optoelectronic and renewable energy applications. PMID- 21643596 TI - Detection of toxic mercury ions using a ratiometric CdSe/ZnS nanocrystal sensor. AB - We have developed a strategy for the ratiometric detection of toxic Hg(2+) ions using a semiconductor nanocrystal energy-transfer donor coupled to a mercury sensitive "turn-on" dye acceptor. The results demonstrate a new paradigm of toxic metal sensing that resolves the difficulties with the use of semiconductor nanotechnology for this purpose. PMID- 21643597 TI - Light driven reactions of single physisorbed azobenzenes. AB - We present a successful attempt of decoupling a dye molecule from a metallic surface via physisorption for enabling direct photoisomerization. Effective switching between the isomers is possible by exposure to UV light via the rotation pathway. PMID- 21643598 TI - Divergent reactivity of alpha-oximino carbenoids: facile access to 2-isoxazolines and 2H-azirines. AB - Mild catalytic reaction conditions for the synthesis of 2-isoxazolines and 2H azirines have been developed via carbenoids derived from alpha-oximino diazo compounds. This has been utilized in the one-pot synthesis of pyrroles in the presence of 1,3-dicarbonyl compounds. PMID- 21643599 TI - Mesoscopic organic nanosheets peeled from stacked 2D covalent frameworks. AB - Novel mesoscopic organic nanosheets were developed by functionalizing bulk 2D organic covalent framework polymers with small molecules. The water-soluble fluorescent nanosheets are promising as nanocarriers for biological applications. PMID- 21643600 TI - Predictive measure of quality of micromixing. AB - We introduce a predictive measure of micromixing termed quantitative overlap (QO). QO depends on the distribution of reactants throughout the reactor and can be calculated by solving equations of diffusion. We used a bimolecular reaction and a capillary microreactor to experimentally prove that QO is proportional to the product yield. PMID- 21643601 TI - One small step in synthesis, a big leap in charge mobility: diphenylethenyl substituted triphenylamines. AB - Star-shaped charge transporting materials with a triphenylamine core and a varying number of diphenylethenyl sidearms, obtained using a one step synthesis procedure from commercially available and relatively inexpensive starting materials and possessing comparatively high hole drift mobility (up to 0.017 cm(2) V(-1) s(-1)), are reported. PMID- 21643602 TI - Formation mechanism of 2-methyl-2-buten-1,4-diol and 2-methyl-3-buten-1,2-diol from 2-methyl-1,3-butadiene on a head-to-head pivalamidato-bridged cis diammineplatinum(III) binuclear complex. AB - Reactions of a pivalamidato-bridged head-to-head (HH) platinum(III) binuclear complex with 2-methyl-1,3-butadiene (isoprene) and p-styrenesulfonate and of an alpha-pyrrolidonato-bridged HH platinum(III) binuclear complex with p styrenesulfonate were studied kinetically using UV-vis spectrophotometry and (1)H NMR spectroscopy, and detailed reaction mechanisms are proposed. Pt(III) binuclear complexes react with p-styrenesulfonate in four successive steps with mechanisms similar to that for an HH alpha-pyridonato-bridged Pt(III) binuclear complex with p-styrenesulfonate. In the case of isoprene, four steps were observed on the basis of UV-vis spectrophotometry. However, the reaction kinetics for steps 1 and 2 correspond to those for the previous reaction system, and those for steps 3 and 4 do not correspond to those for the previous system or to those observed by using (1)H NMR spectroscopy for the present isoprene system. By using UV-vis spectrophotometry, it was shown that isoprene preferentially pi coordinates to the Pt(N(2)O(2)) atom via the double bond adjacent to the methyl group in step 1. In step 2, a second isoprene molecule pi-coordinates to the Pt(N(4)) atom, which is the rate-determining step, followed by nucleophilic attack of a water molecule on the pi-coordinated isoprene on the Pt(N(2)O(2)) atom to form two isomeric sigma-complexes. In the same step, pi-coordinated isoprene on the Pt(N(4)) atom of the sigma-complexes is released. This is different from the reaction of the Pt(III) binuclear complexes with other olefins. In step 3, reductive elimination of the sigma-complexes occurs to form two diols and the HH pivalamidato-bridged Pt(II) binuclear complex. Finally, acid decomposition of the Pt(II) binuclear complex occurs to form monomers in step 4. From (1)H NMR spectroscopic observations, fast isomerization between sigma complexes and reductive elimination of the sigma-complexes occurs in step 3, and isomerization from a 1,4-diol to a 1,2-diol occurs in step 4. PMID- 21643603 TI - Spin crossover in di-, tri- and tetranuclear, mixed-ligand tris(pyrazolyl)methane iron(II) complexes. AB - A series of polynuclear mixed-ligand tris(pyrazolyl)methane iron(II) complexes displaying high temperature spin crossover behaviour has been synthesised. These complexes are of the type [(Fe((3,5-Me(2)pz)(3)CH))(n)(MU-L)](BF(4))(2n), where MU-L is one of five bridging ligands X(CH(2)OCH(2)C(pz)(3))(n), (X = the central linking moiety, pz = pyrazolyl ring and n = 2 (ditopic), 3 (tritopic) or 4 (tetratopic)). Throughout the series the terminal tris(3,5 dimethylpyrazolyl)methane co-ligand (3,5-Me(2)pz)(3)CH and the BF(4)(-) counter anion were kept constant while variations in the central linking moiety have produced three dinuclear complexes and a trinuclear and tetranuclear complex, all isolated as solvates. The three dinuclear complexes are a 1,4-xylene-bridged complex 1.2DME, a 2,6-naphthalene-bridged complex 2.2.5MeCN.2DME and a 1,4-butene bridged complex 3.2DME. The trinuclear complex 4.solvent (solvent undefined) has a 1,3,5-mesitylene core and the tetranuclear complex, 5.8MeCN.2(t)BuOMe, has a 1,2,4,5-tetramethylbenzene core (DME = dimethoxyethane, (t)BuOMe = tertiarybutyl methylether). The trinuclear cluster has a "3-up" cup shape with the cups arranging themselves in pairs to form capsules that contain anion guests. All the solvated compounds have been structurally characterised and both the solvated and desolvated versions have had their magnetic and thermal properties thoroughly investigated by variable temperature magnetic susceptibility, differential scanning calorimetric and Mossbauer spectral methods. They all display typical low spin iron(II) magnetic behaviour at room temperature and all undergo a spin state transition to high spin iron(II) above room temperature. In particular, complex 1.2DME shows an abrupt spin transition which shifts, upon desolvation, to a lower value of T(1/2) and in addition displays a small thermal hysteresis. PMID- 21643604 TI - Low-temperature solid state synthesis and in situ phase transformation to prepare nearly pure cBN. AB - Cubic boron nitride (cBN) is synthesized by a low-temperature solid state synthesis and in situ phase transformation route with NH(4)BF(4), B, NaBH(4) and KBH(4) as the boron sources and NaN(3) as the nitrogen source. Furthermore, two new strategies are developed, i.e., applying pressure on the reactants during the reaction process and introducing the structural induction effect. These results reveal that the relative contents of cBN are greatly increased by applying these new strategies. Finally, almost pure cBN (~90%) crystals are obtained by reacting NH(4)BF(4) and NaN(3) at 250 degrees C and 450 MPa for 24 h, with NaF as the structural induction material. The heterogeneous nucleation mechanism can commendably illuminate the structure induction effect of NaF with face center cubic structure. In addition, the induction effect results in the cBN nanocrystals presenting obvious oriented growth of {111} planes. PMID- 21643605 TI - A regio- and stereoselective synthesis of trisubstituted alkenes via gold(I) catalyzed hydrophosphoryloxylation of haloalkynes. AB - A new stereoselective synthesis of trisubstituted alkenes is developed. Hydrophosphoryloxylation of haloalkynes provides Z-alkenyl halophosphates, which undergo Pd-catalyzed consecutive cross-coupling reactions to afford regio- and stereodefined trisubstituted alkenes. PMID- 21643606 TI - Trans-selective hydrogermylation of alkynes promoted by methyliron and bis(germyl)hydridoiron complexes as a catalyst precursor. AB - Catalytic trans-selective hydrogermylation of terminal and internal alkynes was attained by a methyliron complex, CpFe(CO)(2)(Me), and a bis(germyl)hydridoiron(IV) complex as a catalyst precursor. The structures of (Z) triphenyl-(2-phenylethenyl)germane and the bis(germyl)hydridoiron(IV) complexes CpFe(CO)(H)(GeR(3))(2) (R = Et, Ph) were confirmed by single crystal X-ray diffraction studies. PMID- 21643607 TI - Non-Cp titanium alkoxide-based homolytic ring-opening of epoxides by an intramolecular hydrogen abstraction in beta-titanoxy radical intermediates. AB - A low-valent titanium species derived in situ from Ti(O-i-Pr)(4), Me(3)SiCl and Mg powder in tetrahydrofuran reacted with epoxides to selectively provide less hindered alcohols via a homolytic ring-opening of epoxides, in which the intermediate beta-titanoxy radical intramolecularly abstracted a hydrogen atom from an alkoxy moiety in the titanium complexes. PMID- 21643608 TI - Synthesis and hydroamination catalysis with 3-aryl substituted pyrrolyl and dipyrrolylmethane titanium(IV) complexes. AB - Using an iridium-catalyzed borylation/Suzuki-Miyaura coupling sequence several 3 aryl-pyrroles were accessed; in addition, dipyrrolylmethanes incorporating these 3-arylpyrroles were synthesized. Using the monodentate pyrrolyls, Ti(NMe(2))(2)(HNMe(2))(pyr(3,5-CF3))(2) (1) and a 2,4-diarylpyrrolyl complex Ti(NMe(2))(3)(pyr(Ar/Ar')) (2) were prepared and structurally characterized. Titanium species bearing the new dipyrrolylmethane ligands Ti(NMe(2))(2)(NHMe(2))(dpm(3,5-CF3)) (3) and Ti(NMe(2))(2)(NHMe(2))(dpm(F3)) (4) were also generated. Kinetics under pseudo-first order conditions with 3 and 4 showed them to be measurably more active than the parent derivative without the electron-withdrawing aryl groups. PMID- 21643609 TI - Reactivity of the 16e (p-cymene)Ru half-sandwich complex containing a chelating 1,2-dicarba-closo-dodecaborane-1,2-dithiolate ligand towards diynes. AB - The reactions of the 16e half-sandwich complex (p-cymene)Ru(S(2)C(2)B(10)H(10)) (Ru16e) with 1,4-diethynylbenzene (L1), 3',6-diethynyl-1,1'-binaphthyl-2,7'-diyl diacetate (L2), 2-bromo-5-ethynylthiophene (L3) and 2,5-diethynylthiophene (L4) lead to 18e mononuclear complexes (p-cymene)Ru(S(2)C(2)B(10)H(9))(H(2)CCPhC=CH) (1), (p-cymene)Ru(S(2)C(2)B(10)H(9))[H(2)CC(C(24)H(16)O(4))C=CH] (2), (p cymene)Ru(S(2)C(2)B(10)H(9)) [H(2)CC(C(4)H(2)S)Br] (3) and (p cymene)Ru(S(2)C(2)B(10)H(9)) [H(2)CC(C(4)H(2)S)C=CH] (4), respectively. In all of them, metal-induced B-H activation has occurred, which leads to a stable Ru-B bond, and the structures take a cisoid arrangement. Only in the case of L4, the binuclear complexes [(p-cymene)Ru(S(2)C(2)B(10)H(9))](2)[H(2)CC(C(4)H(2)S)CCH(2)] (5a and 5b) are observed, which are conformational isomers generated by the differing orientations of the p-cymene unit. 4 can be readily converted to the complex (p-cymene)Ru(S(2)C(2)B(10)H(9))[H(2)CC(C(4)H(2)S)COCH(3)] (6) in the presence of silica and H(2)O. All of these products 1-6 were characterized by NMR, IR, elemental analysis and mass spectrometry. The structures of 1, 3, and 5a were also determined by single-crystal X-ray diffraction analysis. PMID- 21643610 TI - Diphosphanes derived from phobane and phosphatrioxa-adamantane: similarities, differences and anomalies. AB - The homodiphosphanes CgP-PCg (1) and PhobP-PPhob (2) and the heterodiphosphanes CgP-PPhob (3), CgP-PPh(2) (4a), CgP-P(o-Tol)(2) (4b), CgP-PCy(2) (4c), CgP P(t)Bu(2) (4d), PhobP-PPh(2) (5a), PhobP-P(o-Tol)(2) (5b), PhobP-PCy(2) (5c), PhobP-P(t)Bu(2) (5d) where CgP = 6-phospha-2,4,8-trioxa-1,3,5,7 tetramethyladamant-9-yl and PhobP = 9-phosphabicyclo[3.3.1]nonan-9-yl have been prepared from CgP(BH(3))Li or PhobP(BH(3))Li and the appropriate halophosphine. The formation of 1 is remarkably diastereoselective, with the major isomer (97% of the product) assigned to rac-1. Restricted rotation about the P-P bond of the bulky meso-1 is detected by variable temperature (31)P NMR spectroscopy. Diphosphane 3 reacts with BH(3) to give a mixture of CgP(BH(3))-PPhob and CgP PPhob(BH(3)) which was unexpected in view of the predicted much greater electron richness of the PhobP site. Each of the diphosphanes was treated with dimethylacetylene dicarboxylate (DMAD) in order to determine their propensity for diphosphination. The homodiphosphanes 1 and 2 did not react with DMAD. The CgP containing heterodiphosphanes 4a-d all added to DMAD to generate the corresponding cis alkenes CgPCH(CO(2)Me)=CH(CO(2)Me)PR(2) (6a-d) which have been used in situ to form chelate complexes of the type [MCl(2)(diphos)] (7a-d) where M = Pd or Pt. The PhobP-containing heterodiphosphanes 3 and 5a-d react anomalously with DMAD and do not give the products of diphosphination. The X-ray crystal structures of the diphosphanes 2, 3, 4a, and 5a, the monoxide and dioxide of diphosphane 1, and the platinum chelate complex 7c have been determined and their structures are discussed. PMID- 21643611 TI - Luminescent square-planar platinum(II) complexes with tridentate 3-bis(2 pyridylimino)isoindoline and monodentate N-heterocyclic ligands. AB - A series of platinum(II) complexes with 1,3-bis(2-pyridylimino)isoindoline (BPI) derivatives were prepared by substitution of the coordinated Cl in the precursor complex Pt(BPI)Cl with a N-heterocyclic ligand such as pyridine, phthalazine or phenanthridine. These complexes display orange to red luminescence in fluid dichloromethane solutions and in the solid states at room temperature. The photophysical properties were tuned by introducing electron-withdrawing -NO(2) or electron-donating -NH(2) to the BPI ligand. The DFT computational studies suggest that the emission in the N-heterocyclic ligand substituted platinum(II) complexes originates mainly from the (3)[pi->pi*(BPI)] (3)IL triplet excited state, mixed with some (3)[dpi(Pt)->pi*(BPI)] (3)MLCT character. Compared with the precursor Pt(BPI)Cl, both the low-energy absorption and the emission in the N-heterocyclic ligand substituted platinum(II) complexes exhibits a distinct blue-shift due to an obviously enhanced contribution from the (3)IL state and a reduced (3)MLCT character. PMID- 21643612 TI - Quasi-linear dependence of cation filling on the photocatalysis of A(x)BO3-based tunnel compounds. AB - A series of A(x)BO(3)-based oxides, (A1)(4)(A2)(2)(A3)(4)Nb(10)O(30) (A1 = Sr, Ba; A2 = Na, K; A3 = ?) as deficient perovskites, ATiO(3) (A = Mg, Ca, Sr, Ba) as ideal perovskites are prepared by solid-state reaction method, and investigated as novel tunnel photocatalysts. The efficiency exhibits a high dependence on the cation filling in the respective tunnels of these compounds. Generally, lower cation filling contributes to a higher activity. The cation filling is rationalized by the crystal packing factor (PF), the photocatalytic efficiency decreases quasi-linearly with PF. Moreover, the linear correlation also exists in the literature involving tunnel photocatalysts of Bi(2)RNbO(7) (R = Y, rare earth element), MIn(2)O(4) (M = Ca, Sr, Ba), ATaO(3) (A = Li, Na, K), etc. Such linear correlation of photocatalysis with the crystal structure has not been reported in the literature. The study of structure-modification by controlling cation-filling for improving activity sheds light on the mechanism of photocatalysis and on developing new high-performance photocatalysts. PMID- 21643613 TI - Soil carbon storage as influenced by tree cover in the Dehesa cork oak silvopasture of central-western Spain. AB - The extent of carbon (C) stored in soils depends on a number of factors including soil characteristics, climatic and other environmental conditions, and management practices. Such information, however, is lacking for silvopastoral systems in Spain. This study quantified the amounts of soil C stored at various depths (0 25, 25-50, 50-75, and 75-100 cm) under a Dehesa cork oak (Quercus suber L.) silvopasture at varying distances (2, 5, and 15 m) to trees. Soil C in the whole soil and three soil fractions (<53, 53-250, and 250-2000 MUm) was determined. Results showed soil depth to be a significant factor in soil C stocks in all soil particle sizes. Distance to tree was a significant factor determining soil C stocks in the whole soil and the 250-2000 MUm soil fraction. To 1 m depth, mean total C storage at 2, 5, and 15 m from cork oak was 50.2, 37, and 26.5 Mg ha(-1), respectively. Taking into account proportions of land surface area containing these C stocks at varying distances to trees to 1 m depth, with a tree density of 35 stems ha(-1), estimated landscape soil C is 29.9 Mg ha(-1). Greater soil C stocks directly underneath the tree canopy suggest that maintaining or increasing tree cover, where lost from disease or management, may increase long term storage of soil C in Mediterranean silvopastoral systems. The results also demonstrate the use of soil aggregate characteristics as better indicators of soil C sequestration potential and thus a tool for environmental monitoring. PMID- 21643614 TI - Recent advances in the transition metal-catalyzed twofold oxidative C-H bond activation strategy for C-C and C-N bond formation. AB - The direct functionalization of heterocyclic compounds has emerged as one of the most important topics in the field of metal-catalyzed C-H bond activation due to the fact that products are an important synthetic motif in organic synthesis, the pharmaceutical industry, and materials science. This critical review covers the recent progresses on the regioselective dehydrogenative direct coupling reaction of heteroarenes, including arylation, olefination, alkynylation, and amination/amidation mainly utilizing transition metal catalysts (113 references). PMID- 21643615 TI - Photophysical and electrochemical properties of Ru(II) complexes containing tridentate bisphosphino-oligothiophene ligands. AB - Nine Ru(II) complexes containing the conjugated oligothiophene ligands 3,3'' bis(diphenylphosphino)-2,2':5',2''-terthiophene (P(2)T(3)) and 4',3''' bis(diphenylphosphino)-3,3''''-dihexyl- 2,2':5',2'':5'',2''':5''',2'''' pentathiophene (P(2)T(5)) were prepared and characterized. P(2)T(3) and P(2)T(5) bond as tridentate ligands and three of the complexes (1, 2 and 5) form green five-coordinate Ru(II) complexes in solution. Cyclic voltammetry, variable temperature UV-vis spectroscopy and time-resolved transient absorption spectroscopy were used to characterize the electronic properties of the complexes. Increased conjugation in the complexes containing the P(2)T(5) ligand resulted in a lowering of the oxidation potential of the oligothiophene, but electropolymerization was not observed. The electronic spectra were dominated by pi-pi* transitions. All of the complexes were non-emissive both at room temperature and low temperature, indicating the excited state decays by other, non-radiative pathways. The transient absorption spectrum of complex 7 shows a species with a band at 475 nm and a lifetime of ~100 ns, assigned to a ligand based triplet state. PMID- 21643616 TI - The first solid phase synthesis of pincer palladium complexes. AB - Both dimeric MU-chlorine bridged and monomeric bidentate Pd(II) complexes with SCN hybrid pincer-type ligands, bearing thiophosphoryl group and imine moiety of the benzothiazole ring as coordination arms, formed in the reaction with (PhCN)(2)PdCl(2) under kinetic control (20 degrees C, dichloromethane solution) were readily converted into the corresponding SCN pincer complexes via solid phase synthesis (neat, 200 degrees C, 15 min). The synthesis of pincer complexes can be performed also by heating (200 degrees C, 5 min) of a homogeneous mixture of the initial reactants, namely, the ligand and (PhCN)(2)PdCl(2), obtained by manual grinding in a mortar. The efficacy of solid phase approaches is comparable with the analogous synthesis in solutions under severe conditions. PMID- 21643617 TI - The CdCl2 effects on synthetic DNAs encaged in the nanodomains of a cationic water-in-oil microemulsion. AB - The present work is dedicated to the study of the interactions of CdCl(2) with the synthetic polynucleotides polyAT and polyGC confined in the nanoscopic aqueous compartment of the water-in-oil microemulsion CTAB/pentanol/hexane/water, with the goal to mimic in vitro the situation met by the nucleic acids in vivo. In biological structures, in fact, very long strings of nucleic acids are segregated into very small compartments having a radius exceedingly smaller than the length of the encapsulated macromolecule. For comparison, the behaviour of polyGC was also studied in aqueous solutions of matched composition. The conformational and thermal stabilities of both polynucleotides enclosed in the inner compartment of the microemulsion are scarcely affected by the presence of CdCl(2), whereas in solution immediate and large effects were observed also at room temperature. The lack of effects of CdCl(2) on the properties of the biopolymers entrapped in the aqueous core of the microemulsion has been attributed to the peculiar characteristics of the medium (low dielectric constant, in particular) which cause a total repression of the CdCl(2) dissociation that is not complete even in water. In fact, several of the numerous effects of CdCl(2) observed on the conformational stability of polyGC in aqueous solutions have also been ascribed to the limited dissociation of the cadmium salt. PMID- 21643618 TI - Existence of time-dependent density-functional theory for open electronic systems: time-dependent holographic electron density theorem. AB - We present the time-dependent holographic electron density theorem (TD-HEDT), which lays the foundation of time-dependent density-functional theory (TDDFT) for open electronic systems. For any finite electronic system, the TD-HEDT formally establishes a one-to-one correspondence between the electron density inside any finite subsystem and the time-dependent external potential. As a result, any electronic property of an open system in principle can be determined uniquely by the electron density function inside the open region. Implications of the TD-HEDT on the practicality of TDDFT are also discussed. PMID- 21643619 TI - Palladium-catalysed hydroxylation and alkoxylation. AB - The formation of oxygen-carbon bonds is one of the fundamental transformations in organic synthesis. In this regard the application of palladium-based catalysts has been extensively studied during recent years. Nowadays it is an established methodology and the success has been proven in manifold synthetic procedures. This tutorial review summarizes the advances on palladium-catalysed C-O bond formation, means hydroxylation and alkoxylation reactions. PMID- 21643620 TI - Ultrafast room temperature NH3 sensing with positively gated reduced graphene oxide field-effect transistors. AB - Reduced graphene oxide (R-GO) under a positive gate potential (n-type conductance) exhibits an instantaneous response and fast recovery for NH(3) sensing, far superior to the performance in p-mode at zero/negative gate potential. Our findings have important implications for fast, repeatable, room temperature gas detection using graphene/R-GO. PMID- 21643621 TI - Prediction of countershock success in patients using the autoregressive spectral estimation. AB - OBJECTIVES: Ventricular fibrillation (VF) is a life-threatening cardiac arrhythmia and within of minutes of its occurrence, optimal timing of countershock therapy is highly warranted to improve the chance of survival. This study was designed to investigate whether the autoregressive (AR) estimation technique was capable to reliably predict countershock success in VF cardiac arrest patients. METHODS: ECG data of 1077 countershocks applied to 197 cardiac arrest patients with out-of-hospital and in-hospital cardiac arrest between March 2002 and July 2004 were retrospectively analyzed. The ECG from the 2.5 s interval of the precountershock VF ECG was used for computing the AR based features Spectral Pole Power (SPP) and Spectral Pole Power with Dominant Frequency weighing (SPPDF) and Centroid Frequency (CF) and Amplitude Spectrum Area (AMSA) based on Fast Fourier Transformation (FFT). RESULTS: With ROC AUC values up to 84.1% and diagnostic odds ratio up to 19.12 AR based features SPP and SPPDF have better prediction power than the FFT based features CF (80.5%; 6.56) and AMSA (82.1%; 8.79). CONCLUSIONS: AR estimation based features are promising alternatives to FFT based features for countershock outcome when analyzing human data. PMID- 21643622 TI - Understanding the causal relations between psychosocial factors at work and health - a circular process. PMID- 21643623 TI - The otoferlin interactome in neurosensory hair cells: significance for synaptic vesicle release and trans-Golgi network (Review). AB - Sound perception in terrestrial vertebrates relies on a structure in the inner ear consisting of the utriculus, sacculus and lagena. In mammals, the lagena has developed into the cochlea where mechanotransduction at ciliated cells leads to ion influx via regulated ion channels. To maintain proper Ca2+ concentration many cellular systems use a variety of functional proteins; the neurosensory systems use calcium-sensors like hippocalcin, visinin or recoverin. In cochlear hair cells the 230 kDa protein otoferlin has been suggested to play this role. While several observations support this hypothesis additional data argue for a more expanded functional profile of otoferlin. Evidence for otoferlin's multiple roles and newer results on otoferlin's interacting partners are presented and the existence of a protein complex as a functional unit ('interactome') in the cochlea and further tissues is suggested. PMID- 21643624 TI - Opioids increase bladder cancer cell migration via bradykinin B2 receptors. AB - Data relating opioid treatment and modification of cancer cell migration (a prerequisite of metastasis) both in vitro and in vivo are diverging. In the present report we show that opioids increase the migratory activity of bladder cancer cells (T24 and EJ) and we provide a new mechanistic insight, explaining (at least partially) their action: we report that the enhanced opioid-related cell migration is controlled (in the absence of opioid receptors) through their interaction with bradykinin B2 receptors. Indeed, in these cell lines, opioids increase migration, adhesion, spreading and invasion by re-arranging actin cytoskeleton, increasing MMP-2 and -9 secretion and triggering specific intracellular signaling cascades in a non-opioid receptor mediated manner. An interaction, albeit with low affinity, of opioids with the bradykinin B2 receptor is reported, resulting in the increase of migration, while B2 antagonists revert this action. A systematic assay of different human epithelial cancer cell lines confirmed that only the B2-positive/opioid receptor-negative bladder cancer cells present this opioid-related increased migration/invasive phenotype. We suggest that opioid administration in cancer patients should be re-evaluated, keeping in mind that they may have other beneficial (protection) or adverse effects (spreading of cancer cells), in spite of their unique role in pain relief. PMID- 21643625 TI - Proteolysis-inducing factor core peptide mediates dermcidin-induced proliferation of hepatic cells through multiple signalling networks. AB - Dermcidin is a candidate oncogene capable of increasing the number of cultured neuronal, breast cancer and prostate cancer cells and improving the survival of hepatic cells. The dermcidin gene encodes the proteolysis-inducing factor core peptide (PIF-CP) and the skin antimicrobial peptide DCD-1. The peptide responsible for inducing proliferation of cells and the mechanisms involved are unknown. In this study, we confirmed a proliferative effect of dermcidin overexpression of 20% (p<0.02) in the HuH7 human hepatic cell line. Proliferation was abrogated by prevention of PIF-CP translation or inactivation of its calcineurin-like phosphatase domain by site-directed mutagenesis. Prevention of DCD-1 translation had no effect. Treatment of cells with a 30 amino acid synthetic PIF-CP induced an analogous increase in proliferation of 14%. Microarray analysis of PIF-CP-treated cells revealed low but significant changes in 111 potential mediator genes. Pathway analysis revealed several gene networks involved in the cellular response to the peptide, one with VEGFB as a hub and two other networks converging on FOS and MYC. Quantitative PCR confirmed direct upregulation of VEGFB. These data reveal PIF-CP as the key mediator of dermcidin induced proliferation and demonstrate induction of key oncogenic pathways. PMID- 21643626 TI - Expression of HGF and VEGF in the cerebral tissue of adult rats with chronic hydrocephalus after subarachnoid hemorrhage. AB - The hepatocyte growth factor (HGF) and vascular endothelial growth factor (VEGF) are important cytokines with modulatory actions in the nervous system. The present study aimed to investigate the role and expression of HGF and VEGF in the cerebral tissue of adult rats with chronic hydrocephalus after subarachnoid hemorrhage. Adult female Wistar rats were randomly divided into 4 groups: a control group (n=20) and 3 experimental subgroups (n=60). Subarachnoid hemorrhage was induced by the injection of 0.4 ml of non-heparinized autologous arterial blood into the cisterna magna of experimental animals on day 0 with a second injection 2 days later. The rats were sacrificed within 24 h of magnetic resonance imaging (MRI) examination at 2, 4, or 6 weeks. The excised brains were studied by RT-PCR, immunohistochemical and Western blot analyses as we examined HGF and VEGF mRNA and protein expression. Chronic hydrocephalus was induced in 21 rats after subarachnoid hemorrhage. After 2 weeks, the expression of HGF and VEGF in the cerebral tissue was significantly increased in the experimental group compared to the controls, especially in periventricular white matter. Our results indicate that HGF and VEGF participate in the pathological injury and repair of cerebral tissue in rats with chronic hydrocephalus after subarachnoid hemorrhage. PMID- 21643627 TI - Celecoxib attenuates liver steatosis and inflammation in non-alcoholic steatohepatitis induced by high-fat diet in rats. AB - Cyclooxygenase-2 (COX-2) is involved in the process of non-alcoholic steatohepatitis (NASH). However, the role of the COX-2 inhibitor in NASH has not yet been elucidated. Therefore, in the present study, we investigated the role of celecoxib in a rat model of NASH induced by a high-fat diet (HFD). Wistar rats were administered HFD by gavage, and rats administered normal saline by gavage served as the controls. After 4 weeks of HFD feeding, the rats were treated with celecoxib (20 mg/kg/day) or placebo for 4 weeks. At the end of 4 and 8 weeks, histological changes in the livers of the rats were analyzed using hematoxylin and eosin; blood was collected to detect biochemical indicators (serum aminotransferase and triglyceride). Liver triglyceride content was measured using the triglyceride E-test kit. The liver expression of COX-2, nuclear factor-kappa enhancer binding protein (NF-kappaB) subunits p50 and p65 was measured by real time reverse transcription-polymerase chain reaction and/or Western blotting. Infiltration of steatosis and inflammation in cells was observed in the livers after 4 weeks of HFD administration, and marked steatosis and inflammation was induced after 8 weeks. These histological changes were significantly attenuated after celecoxib treatment. Reduced serum alanine aminotransferase and triglyceride (TG) levels and TG content in the liver were observed in the HFD rats that received celecoxib. Moreover, celecoxib suppressed hepatic COX-2 messenger RNA and protein expression. The NF-kappaB subunit p50 and p65 protein levels in the HFD rats were also attenuated after celecoxib treatment. The results indicate that the induction of COX-2 occurs in association with NF-kappaB activation in HFD-induced NASH rats. Celecoxib may protect against the development of steatohepatitis induced by HFD. PMID- 21643628 TI - Detection of SRS19-6MuLV in mouse dendritic cell sarcoma and its tumorigenesis. AB - SRS19-6MuLV is a member of the MuLV family originally isolated from the Tianjin Shanghai-Zunyi complex of murine leukemia. A notable characteristic of this virus is that it induces tumors of multiple hematopoietic lineages, including myeloid, erythroid, T-lymphoid and B-lymphoid. In a previous study, a sequence with high homology to SRS19-6MuLV in a murine dendritic cell sarcoma (DCS) was identified through cDNA expression screening with mAb 983D4. To investigate the relationship between SRS19-6MuLV and DCS, the existence of a specific SRS19-6MuLV DNA fragment in DCS cells, 15 murine tumor cells, 2 murine tumor tissues, 12 normal murine cells/tissues, 11 human tumor cell lines and SRSV/3T3 (NIH/3T3 cells infected with SRS cell supernatant) was detected by PCR. The specific fragment of SRS19 6MuLV was detected in DCS, mouse fore-gastric cancer cells, LII tumor tissue from which DCS is derived and SRSV/3T3. In addition, the integration sites of SRS19 6MuLV in the positive cells were examined by inverse PCR. Thus, 7 integration sites for SRS19-6MuLV were detected in DCS and 3 in SRSV/3T3. Analysis of sequences by BLAST revealed that some of the integration sites were associated with common fragile sites and some Ras-regulating miRNAs. Our results indicate that SRS19-6MuLV not only induced four types of leukemia, but also induced DCS. This virus does not infect human cells. Multiple integration of SRS19-6MuLV into chromosomes around fragile sites accounts for its carcinogenic effects. PMID- 21643629 TI - Overexpression of DEP domain containing mTOR-interacting protein correlates with poor prognosis in differentiated thyroid carcinoma. AB - DEP domain containing mammalian target of rapamycin (mTOR)-interacting protein (DEPTOR), a recently identified mTOR-interacting protein, is a novel candidate oncogene. Previous studies reveal that high DEPTOR expression is required to maintain PI3K and Akt activation and to inhibit apoptosis. However, its significance in differentiated thyroid carcinoma (DTC) is not yet known. The present study verifies the mRNA and protein expression of DEPTOR in five cell lines, DTC tissues and normal adjacent tissues. Tissue microarrays of 114 DTC patients were used to detect DEPTOR protein expression. The assessment of DEPTOR levels demonstrated that DEPTOR in DTC cells and tissues was significantly increased compared with normal cells and adjacent normal tissues. DEPTOR protein expression was significantly associated with lymph node status, extrathyroid extension and distant metastasis. Patients exhibiting high DEPTOR expression were statistically susceptible to earlier recurrence and poorer survival than those with low expression. Univariate and multivariate analyses showed that DEPTOR expression was an independent prognostic factor for DTC recurrence. In conclusion, our data indicate DEPTOR as a novel prognostic marker for DTC. PMID- 21643630 TI - Interobserver variability of dynamic MR imaging of the temporomandibular joint. AB - PURPOSE: This study investigated the interobserver variability of dynamic magnetic resonance (MR) imaging of the temporomandibular joint (TMJ). MATERIALS AND METHODS: Sixty-two patients with internal derangement of the TMJ (124 TMJs) were investigated with a 1.5-T MR imaging system during physiological opening and closing of the mouth. Two readers evaluated independently the quality of the dynamic examination (Q: nondiagnostic, diagnostic, optimal), condylar motion (CM: limited, suboptimal, optimal), condylar orientation (CO: in-plane, through-plane shift), disc visibility and movement (DV: visible, nonvisible; DM: normal, reducing, nonreducing dislocation) and joint effusion (JE: present, absent). For each TMJ, the condylar path was measured by tracing the position of the condyle in the frames of the dynamic acquisition. Agreement between the two readers was assessed with Cohen's Kappa and the Bland-Altman method. RESULTS: Interobserver agreement was almost perfect for Q (nondiagnostic, diagnostic, optimal: 0.8%, 4.9%, 94.3%; kappa=1), CM (limited, suboptimal, optimal: 14%, 26.4%, 59.5%; kappa=0.84) and DV (visible, nonvisible: 100%, 0%). Substantial agreement was found for DM (normal, reducing, nonreducing: 66.1%, 14.8%, 19.1%; kappa=0.64) and JE (present, absent: 41.3%, 58.7%; kappa=0.67). Moderate agreement was found for CO (in-plane, through-plane shift: 94.2%, 5.8%; kappa=0.41). As for the condylar path, the means of the percentage differences and limits of agreement (LA) were 3% (LA: -34.5%, 28.3%) on the right and -1.2% (LA: -35%, 32.6%) on the left. CONCLUSIONS: In dynamic imaging of the TMJ, qualitative assessment of condyle disc movement and joint effusion is minimally dependent on the reader's evaluation. Measurement of the condylar pathway shows an interobserver variability of +/-30%. PMID- 21643631 TI - Search for compensation postures with videofluoromanometric investigation in dysphagic patients affected by amyotrophic lateral sclerosis. AB - PURPOSE: This study was undertaken to verify the effectiveness of compensatory postures, suggested on the basis of the type of dysphagia identified at videofluoromanometric (VFM) investigation to ensure safe oropharyngeal transit. MATERIALS AND METHODS: Eighty-one patients with amyotrophic lateral sclerosis (ALS) underwent speech therapy assessment and VFM investigation of the swallowing process. In the event of altered transit, penetration or aspiration of contrast material into the airways, compensation postures for correction of the swallowing disorder were suggested and verified during VFM examination. RESULTS: In 37 patients, contrast agent transport was preserved and safe; in 19, we observed penetration of the contrast agent into the laryngeal inlet without aspiration; in 24, there was aspiration (four preswallowing, eight intraswallowing, nine postswallowing, three mixed), whereas in one patient no transit was seen. Penetration without aspiration was resolved by coughing or throat clearing; aspiration was resolved in 13 patients by assuming the chin-tuck posture and in six by rotating the head; in five patients, it was not resolved. A hyperextended head posture proved to be effective to resolve lack of transit. CONCLUSIONS: By correlating morphological with functional data, VFM enables one not only to precisely characterise the dysphagic disorder but also to identify the most appropriate compensation posture for each patient and verify its effectiveness. PMID- 21643632 TI - Ultrasound-guided thermal radiofrequency ablation (RFA) as an adjunct to systemic chemotherapy for breast cancer liver metastases. AB - PURPOSE: The aim of this study was to evaluate the effectiveness of radiofrequency thermal ablation (RFA) of liver metastases from breast cancer and its impact on survival. MATERIALS AND METHODS: Thirteen female patients (age range 36-82 years; median 54.5 years) underwent RFA for the treatment of 21 liver metastases from breast cancer. The procedures were performed under ultrasound (US) guidance using an RF 2000 or RF 3000 generator system and Le Veen monopolar needle electrodes. Follow-up was carried out by computed tomography (CT) after 1, 3, 6 and 12 months. RESULTS: Technical success was 100%. No major or minor complications occurred at the end of the procedure. In our series, 7/21 lesions in 7/13 patients increased in size at 7, 18, 19 and 38 months. This resulted in a mean disease-free interval of 16.6 months. Mean overall survival after RFA was 10.9 months. CONCLUSIONS: RFA appears to be a useful adjunct to systemic chemotherapy and/or hormone therapy in the locoregional treatment of hepatic metastases from breast cancer. RFA may also be a less invasive alternative to surgery in the locoregional treatment of liver metastases from breast cancer. PMID- 21643633 TI - Diagnostic accuracy of 64-slice computed tomography coronary angiography in a large population of patients without revascularisation: registry data in NSTEMI acute coronary syndrome and influence of gender and risk factors. AB - PURPOSE: This study sought to evaluate the diagnostic accuracy of computed tomography coronary angiography (CTCA) for detecting significant coronary artery stenosis (>=50% lumen reduction) compared with conventional coronary angiography (CAG) in non-ST-elevation myocardial infarction-acute coronary syndrome (NSTEMI ACS) and in subgroups selected by gender and number of risk factors (RF). MATERIALS AND METHODS: We selected from a population of 1,500 patients in a multicentre registry with NSTEMI-ACS who had undergone CTCA and CAG, (n=237; 187 men, mean age 63+/-10 years). Diagnostic accuracy and likelihood ratios (LR) of CTCA were assessed against CAG in the total population and subgroups (men, women: 0 RF = absence of RF, 1-2 RF = presence of one or two RF, >2 RF = presence of more than two RF). RESULTS: The prevalence of obstructive disease was 53%. In the per-patient analysis, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of CTCA were 100% (men 100%; women 100%; 0 RF 100%; 1-2 RF 100%; >2 RF 100%), 95% (men 98%; women 50%; 0 RF NA% (NA, not assessable); 1-2 RF 96%; >2 RF 96%), 95% (men 98%; women 91%; 0 RF 91%; 1-2 RF 96%; >2 RF 96%), 100% (men 100%; women 100%; 0 RF NV%; 1-2 RF 100%; >2 RF 100%), respectively. The per-segment analysis showed a reduction in PPV (ranging between 56% and 67%). The per-patient LR+ ranged between 18 and 27, whereas LR-were always 0. We observed no significant differences in diagnostic accuracy between subgroups. CONCLUSIONS: CTCA is a reliable diagnostic modality with high sensitivity and NPV in NSTEMI-ACS patients who are not candidates for early revascularisation, regardless of gender and number of risk factors. PMID- 21643634 TI - Computed tomography coronary angiography in the selection of outlier patients: a feasibility report. AB - PURPOSE: This study evaluated criteria, presence and distribution of outlier patients by means of computed tomography coronary angiography (CTCA) in a large institutional database. MATERIAL AND METHODS: From a population of 2,881 consecutive patients (1,842 men, mean age 62 +/- 13 years) in sinus rhythm who underwent CTCA, we extracted data on patients with suspected coronary artery disease (CAD). We selected patient outliers in the fifth and sixth decades of life with the following criteria: >= 3 risk factors and absence of CAD, zero to one risk factors and >= 5 diseased coronary segments. Diabetes was excluded from risk factors because of the different impact on CAD. RESULTS: The patient population consisted of 2,432 individuals with suspected CAD (1,495 men, age 62 +/- 13 years). The prevalence of obstructive CAD (>= 50% lumen reduction at CTCA) was 36% (863/2,432). Patients with normal coronary arteries accounted for 34% of the total (837/2,432; 431 men, age 55 +/- 14 years). Of these, 210 were in the fifth and 231 in the 6th decade (men 196, women 245); those with >= 3 risk factors accounted for 4.2% of the total (102/2,432; men 42, women 60). Patients with >= 5 diseased coronary segments accounted for 28% of the total (686/2,432; 510 men, age 68 +/- 10 years). Of these, 115 were in the fifth and 270 in the sixth decade (men 309, women 76); those with zero to one risk factors accounted for 3.0% (73/2,432; men 66, women 7). CONCLUSIONS: CTCA is a reliable noninvasive diagnostic modality that can be used to identify outlier patients. This will enable dedicated trials aimed at characterising biomarkers and genomics of protective and nonprotective factors against CAD and its complications. PMID- 21643635 TI - The "blind age assessment": applicability of Greulich and Pyle, Demirjian and Mincer aging methods to a population of unknown ethnic origin. AB - PURPOSE: Age estimation is one of the most crucial issues in case of unknown deceased as well as in the living and is very frequently of radiological interest. Three methods for age estimation have been designated as the most reliable among the others: Greulich and Pyle, Demirjian and Mincer. The literature provides several studies concerning their applicability in different geographic contexts. However, not always can ancestry be ascertained, for example, in the case of badly preserved corpses. In these cases, age assessment must be performed without the corrections suggested by the literature for different ethnic groups. One may therefore wonder how reliable the result of age assessment performed without knowing the racial group to which the subject belongs may be. This study aimed at testing the applicability of the Greulich and Pyle Atlas, the Demirjian and the Mincer methods on a mixed population to compare skeletal and dental methods of age estimation. MATERIALS AND METHODS: X-ray films of 167 subjects aged between 4 and 31 years from more than 18 countries were recruited. One hundred and nine orthopantomographs (OPG) of children aged between 4 and 15.5 years were evaluated by Demirjian's method; whenever the highest Demirjian score was reached (31 cases), the Mincer method was applied. The skeletal maturation of 54 subjects aged between 7 and 19 years was determined by the Greulich and Pyle method. RESULTS: The lowest average variance from chronological age was shown by the Greulich and Pyle method, followed by Demirjian. The Mincer method showed very high mean variances. CONCLUSIONS: Mean variances from the different methods do not significantly differ from data reported in the literature and demonstrate that the reliability of Demirjian, and Greulich and Pyle as they stand may be applied satisfactorily to remains or individuals of unknown ethnic origin. PMID- 21643636 TI - Diagnostic accuracy of 64-slice computed tomography coronary angiography in a large population of patients without revascularisation: registry data on the comparison between male and female population. AB - PURPOSE: This study was undertaken to evaluate the diagnostic accuracy of computed tomography coronary angiography (CTCA) for detecting significant coronary artery stenosis (>=50% lumen reduction) compared with conventional coronary angiography (CAG) in a male and female population. MATERIAL AND METHODS: A total of 1,372 patients (882 men, 490 women; mean age 59.3 +/- 11.9 years) in sinus rhythm imaged with CTCA (64-slice technology) and CAG were enrolled. Diagnostic accuracy and likelihood ratios (LR+ and LR-) of CTCA were assessed against CAG for the male and female populations. RESULTS: The prevalence of obstructive disease was 53% (men 58%; women 43%). CAG demonstrated the absence of significant coronary artery disease (CAD) in 47% (men 42%; women 56%), single vessel disease in 25% (men 36%; women 22%) and multivessel disease in 29% (men 32%; women 23%) of patients. In the per-patient analysis, sensitivity, specificity and positive (PPV) and negative (NPV) predictive values of CTCA were 99% (men 98%; women 100%), 92% (men 92%; women 92%), 94% (men 95%; women 90%) and 99% (men 98%; women 100%), respectively. The per-patient likelihood ratios (LR) in the total population (LR+=12.4 and LR-=0.011), the male (LR+=12.9 and LR =0.016) and female (LR =11.9 and LR-=0) populations were very good. We observed no significant differences in diagnostic accuracy between male and female populations. CONCLUSIONS: CTCA is a reliable diagnostic modality with high sensitivity and NPV in the female population. PMID- 21643637 TI - Synovial sarcoma: CT imaging of a rare primary malignant tumour of the thorax. AB - PURPOSE: This paper presents computed tomography (CT) features of three patients with primary synovial sarcoma of the lung (PSSL) who came to our attention and underwent surgery; reviews of the literature on this rare thoracic tumour are also presented. MATERIALS AND METHODS: The patients, all men, with a mean age of 58 years, underwent clinical and radiological re-evaluation after receiving a histological diagnosis. None of the patients had multifocal disease or other concomitant neoplasms. All patients had undergone both chest X-rays and computed tomography, and two had also been studied with positron emission tomography (PET) CT. Two patients underwent surgical removal of the tumour, whereas the third initially underwent surgery (following an incorrect diagnosis) and then thoracoscopic biopsy of the pleural lesions that subsequently arose. RESULTS: In each case, chest X-rays showed changes, with the presence of pulmonary masses noted in all patients. In one patient, pleural effusion was also visible. CT scans showed parenchymal masses that were largely of a colliquative nature (in two out of three patients). Ipsilateral pleural effusion was present in two patients, associated in one with solid nodules within the pleura. Mediastinal lymphadenopathy, which was not radiologically significant, was present in only one patient. The two patients who also underwent PET-CT examination showed pathological tracer uptake confined to the lesion site without other thoracoabdominal or musculoskeletal localisations. CT-guided biopsy, performed in one patient only, was positive for mesenchymal tumour. In the two patients who underwent surgery, a definitive diagnosis of monophasic synovial sarcoma of the lung was made. The diagnosis of monophasic synovial sarcoma in the third patient was confirmed using thoracoscopic biopsy DISCUSSION: Both in the cases described and in those identified in the literature review, standard chest X-rays mainly showed a parenchymal mass of pleural origin with either irregular or well-defined margins. CT characteristics are more definite, with evidence of a mass with regular and sharply defined margins, occasionally polycyclic, with inhomogeneous density due to the presence of colliquative areas within the tumour. CONCLUSIONS: Although PSSL is a rare tumour, a pulmonary mass of inhomogeneous density, associated with pleural effusion but without lymphadenopathy, detected in an asymptomatic or poorly symptomatic patient, should lead to PSSL being considered in the differential diagnosis, provided that metastases from the more common synovial sarcomas of the musculoskeletal system have been excluded. PMID- 21643638 TI - Renal volume assessment with 3D ultrasound. AB - PURPOSE: This study was undertaken to determine the accuracy of 3D ultrasound (US) in assessing renal volume, with multislice computed tomography (MSCT) considered as the gold standard. MATERIALS AND METHODS: Forty-nine patients (30 men, 19 women; age range 30-82 years) underwent abdominal contrast-enhanced MSCT and 3D-US performed with a 3.5-MHz 3D/4D convex-array probe. The results of the two modalities were compared with the Wilcoxon test. Variability between the two measurements was determined with the Bland-Altman method and reported in terms of bias and coefficient of repeatability (CoR). RESULTS: Mean values obtained were 210 ml with MSCT and 192 ml with 3D-US (p<0.001). Analysis of variability per patient between MSCT and 3D-US showed a bias of 19 ml, a CoR of 47 ml and an accuracy of 78%, with an average 3D-US underestimation of 19 ml (9%). Analysis of variability per kidney showed a bias of 9 ml, a CoR of 34 ml and an accuracy of 80%. CONCLUSIONS: Three-dimensional US is a valuable technique for monitoring renal volume, whereas MSCT may be reserved for assessing renal anatomy and relationships with neighbouring organs. PMID- 21643639 TI - Enlarged lymph nodes in the upper abdomen after liver transplantation: imaging features and clinical significance. AB - PURPOSE: Our aim was to review our single-centre experience regarding imaging features and pathological conditions arising from enlarged lymph nodes following orthotopic liver transplantation (OLD) and identify imaging features that may allow narrowing the differential diagnosis. MATERIALS AND METHODS: Between January 1997 and September 2008, 715 OLTs were performed at our Institution in 585 patients. In 19 patients (2.6 %) ultrasound (US) showed enlarged lymph nodes in the upper abdomen, and abdominal computed tomography (CT) was performed RESULTS: Histological results showed posttransplantation lymphoproliferative disorder (PTLD) in six patients (31.5 %), hepatic tumoral disease in six (31.5 %), disseminated systemic tuberculosis in one (5.2%) and sarcoidosis in one (5.2%). Histological results showed nonspecific reactive lymph node hyperplasia in five patients (26%). CONCLUSIONS: Detecting enlarged lymph nodes in the upper abdomen after liver transplantation is and infrequent occurrence; however, thorough imaging is required to detect and characterise a wide variety of disorders. Most of the time, enlarged lymph nodes will be related to posttransplantation lymphoproliferative disorder or tumoral recurrence. PMID- 21643640 TI - Relationship between body composition, body mass index and bone mineral density in a large population of normal, osteopenic and osteoporotic women. AB - PURPOSE: The knowledge of factors modulating the behaviour of bone mass is crucial for preventing and treating osteoporotic disease; among these factors, body weight (BW) has been shown to be of primary importance in postmenopausal women. Nevertheless, the relative effects of body composition indices are still being debated. Our aim was to analyze the relationship between body mass index (BMI), fat and lean mass and bone mineral density (BMD) in a large population of women. Moreover, this study represents a first important report on reference standard values for body composition in Italian women. MATERIALS AND METHODS: Between 2005 and 2008, weight and height of 6,249 Italian women (aged 30-80 years) were measured and BMI was calculated; furthermore BMD, bone mineral content, fat and lean mass were measured by dual-energy X-ray absorptiometry. Individuals were divided into five groups by decades (group 1, 30.0-39.9; group 2, 40.0-49.9; group 3, 50.0-59.9; group 4, 60.0-69.9; group 5, 70.0-79.9). Differences among decades for all variables were calculated using a one-way analysis of variance (ANOVA) and Bonferroni test by the SPSS programme. RESULTS: Mean BW was 66.8+/-12.1 kg, mean height 159.1+/-6.3 cm and mean BMI 26.4+/-4.7 kg/m(2). According to BW and BMI, there was an increase of obesity with age, especially in women older than 50 years (p<0.001). Lean mass increased until 50 years of age but significantly decreased after this age (p<0.001). The percentage of osteopenia and osteoporosis in the examined population was 43.0% and 16.7%, respectively. CONCLUSIONS: Our data show that obesity significantly decreased the risk for osteoporosis but did not decrease the risk for osteopenia. It is strongly recommended that a strong policy regarding prevention of osteopenia and osteoporosis be commenced. An overall examination of our results suggests that both fat and lean body mass can influence bone mass and that their relative effect on bone could be modulated by their absolute amount and ratio to total BW. PMID- 21643641 TI - Dynamic 4D MR angiography versus multislice CT angiography in the evaluation of vascular hepatic involvement in hereditary haemorrhagic telangiectasia. AB - PURPOSE: Hereditary haemorrhagic telangiectasia (HHT), or Rendu-Osler-Weber disease, is a rare autosomal dominant disorder characterised by mucocutaneous or visceral vascular abnormalities that may be widely distributed throughout the cardiovascular system. The purpose of this study was to compare multislice computed tomography angiography (MSCTA) and 4D dynamic contrast-enhanced magnetic resonance angiography (D-MRA) for evaluating vascular hepatic involvement in patients with HHT. MATERIALS AND METHODS: Fifty-two consecutive HHT patients underwent MSCTA and D-MRA examinations for systematic analysis of vascular visceral involvement. The images from the two techniques were reviewed independently by two expert radiologists to identify the following vascular abnormalities: telangiectases or large vascular masses; perfusion disorders [transient hepatic attenuation differences (THADs)]; hepatic arteriovenous malformations (HAVMs). Data, as well as diameters of the common hepatic artery and portal vein, were compared with Cohen's kappa statistic, Student's t test and receiver operating characteristic (ROC) curve analysis, as appropriate. RESULTS: Both MSCTA and D-MRA detected one or more of the following hepatic vascular abnormalities in 36/52 cases (telangiectases in 29/52, THADs in 23/52 and HAVMs in 25/52[CE1]). A good concordance was found between the two techniques when determining the type of hepatic shunt (kappa=0.9). No statistically significant differences were found when comparing mean common hepatic artery and portal vein diameters (p=0.09 and 0.22, respectively) and their accuracy in predicting HAVMs. CONCLUSIONS: D-MRA has the same diagnostic accuracy as MSCTA and has the advantage of being less invasive due to the absence of ionising radiation. PMID- 21643642 TI - Volumetric analysis of the aneurysmal sac with computed tomography in the follow up of abdominal aortic aneurysms after endovascular treatment. AB - PURPOSE: Our aim was to assess the usefulness of volumetric analysis for the follow-up of abdominal aortic aneurysms after endovascular repair (EVAR) and operator independence of the method. MATERIALS AND METHODS: We retrospectively evaluated 99 computed tomography (CT) exams of 33 patients. Two blinded operators assessed the volume before treatment and after EVAR at 1-3 and 12-24 months. Friedman's statistical test was used to assess the reproducibility of the method. The time required for postprocessing by the two operators was compared. RESULTS: One patient was excluded. Twenty-one patients showed no endoleak: 12/21 showed a volume reduction at both follow-up scans (9.7% and 19.5%, respectively); 8/21 showed an early volume increase (9.8%) with a late reduction (10.5%); 1/21 patient showed a volume increase at both follow-up scans (endotension). Eleven patients had an endoleak (one type I, nine type II and one type III); 4/9 type II endoleaks showed a volume reduction at both post-EVAR scans (8.5% and 19.5%). All other cases showed a volume increase after EVAR (type II 15.4%/16.8%, type I 24.1%/9.1%, type III 8%/10.7%). The Friedman statistical test assessed operator independence with p < 0.001. Mean difference between the two operators was 0.9% (0-4.3%). CONCLUSIONS: CT volume analysis is an accurate and reproducible modality for the follow-up of abdominal aortic aneurysms after EVAR. At early follow-up, contrast-enhanced CT remains mandatory to identify small endoleaks. For later follow-up, volumetric analysis would eliminate the need for contrast material in asymptomatic patients with stable or decreasing aneurysm volume. PMID- 21643643 TI - Association of insulin resistance with arterial stiffness in nondiabetic peritoneal dialysis patients. AB - BACKGROUND: Insulin resistance is a risk factor for cardiovascular morbidity and mortality in the general and end-stage renal disease populations. In this study, we investigated the association between insulin resistance and arterial stiffness in nondiabetic peritoneal dialysis (PD) patients. METHODS: Fifty-three patients were enrolled. Patients were divided into 2 groups as homeostasis model assessment of insulin resistance (HOMA-IR) <= 2.97 (low) and >2.97 (high). Carotid-femoral pulse wave velocity (c-f PWV) analysis and intima-media thickness of the carotid artery were measured. RESULTS: Mean age was 46 +/- 12 years and HOMA-IR was 2.97 +/- 1.77 (0.77-8.88). Mean c-f PWV was 7.6 +/- 1.7 m/s. HOMA-IR was positively correlated with age, body mass index, and c-f PWV and negatively with serum HDL cholesterol and parathormone. In linear regression analysis, age and mean arterial pressure were predictors for c-f PWV. When patients were divided into 2 groups according to median age as <= 49 and >50, mean arterial pressure, male gender, and age were predictors for c-f PWV in patients aged <= 49, whereas HOMA-IR was the only predictor for c-f PWV in patients aged >50 years. CONCLUSION: Insulin resistance is an independent risk factor for arterial stiffness in PD patients older than 50 years. IR is not associated with carotid intima-media thickness. PMID- 21643644 TI - Role of botulinum toxin-A in refractory idiopathic overactive bladder patients without detrusor overactivity. AB - BACKGROUND: To evaluate the efficacy of intradetrusor botulinum toxin-A (BTX-A) in idiopathic overactive bladder patients (OAB) refractory to anti-muscarinic therapy, without detrusor overactivity (DOA) on urodynamics. METHODS: Patients with refractory idiopathic OAB without DOA on urodynamics were prospectively enrolled. Subjects completed a 3-day voiding diary (3-VD), urogenital distress inventory-6 questionnaire (UDI-6) and graded their quality of life on a 10-cm visual analog scale (VAS) prior to study enrollment and at week 12 postinjection. All patients underwent multichannel urodynamics at study enrollment and 12 weeks postinjection. Improvement was analyzed based on 3-VD, UDI-6 questionnaire, VAS scores, and urodynamic parameters at week 12 postinjection compared to study enrollment. RESULTS: The study included 32 patients. Mean +/- SD age was 56 +/- 16. There were 27 women and 5 men. Nineteen patients had OAB without incontinence (OAB-dry) and 13 patients had OAB with incontinence (OAB-wet). In OAB-dry patients, mean +/- SD urinary frequency dropped from 24 +/- 11 to 10 +/- 4 by week 12. In OAB-wet patients, mean +/- SD urge incontinence episodes dropped from 7.9 +/- 5 to 0 +/- 2.6 by week 12. Mean UDI-6 and VAS scores improved significantly in both groups by week 12 (P = 0.0001). On comparing urodynamic parameters, OAB-wet patients showed significant decrease in maximum detrusor pressure during the voiding phase at week 12 compared to baseline values (P = 0.02). CONCLUSIONS: Refractory idiopathic OAB patients without DOA on urodynamics may benefit from intradetrusor BTX-A. PMID- 21643646 TI - Renal reserve in the oldest old: calculation of concerns regarding creatinine clearance rate with cimetidine in the elderly. PMID- 21643645 TI - Endogenous soluble receptor of advanced glycation end-products (esRAGE) is negatively associated with vascular calcification in non-diabetic hemodialysis patients. AB - BACKGROUND: Advanced glycation end-products (AGE) accumulate in CKD and may predispose to cardiovascular disease by inducing inflammatory and oxidant stress in the vascular endothelium. Soluble forms of the receptor for AGE (RAGE) may be protective against these effects by binding AGE in the soluble phase. Accumulating evidence suggests a protective role of soluble RAGE against vascular calcification. This study investigates the association between endogenous soluble RAGE (esRAGE) and vascular calcification in hemodialysis patients. METHODS: We studied 65 non-diabetic hemodialysis patients, on 3 * 4 h dialysis schedule, and 19 controls. Serum levels of esRAGE, hsCRP, parathormone, lipids, calcium, and phosphorus were measured. Aortic calcification index (ACI) was measured using non contrast CT of the abdominal aorta. RESULTS: Aortic calcification was detected in 64 out of 65 hemodialysis patients. Levels of esRAGE were lower in hemodialysis patients (278 pg/ml, SD 101.1) than in controls (443 +/- 109 pg/ml; P = 0.001). ACI correlated negatively in stepwise multivariate analysis with esRAGE (P = 0.002) and positively with hsCRP (<0.0001), systolic blood pressure (P < 0.0001) and dialysis vintage (P = 0.05); R (2) = 0.65. CONCLUSION: Levels of esRAGE were low among hemodialysis patients and correlated negatively with ACI. PMID- 21643647 TI - Glutamatergic input from specific sources influences the nucleus accumbens ventral pallidum information flow. AB - The nucleus accumbens (NAc) is positioned to integrate signals originating from limbic and cortical areas and to modulate reward-related motor output of various goal-directed behaviours. The major target of the NAc GABAergic output neurons is the ventral pallidum (VP). VP is part of the reward circuit and controls the ascending mesolimbic dopamine system, as well as the motor output structures and the brainstem. The excitatory inputs governing this system converge in the NAc from the prefrontal cortex (PFC), ventral hippocampus (vHC), midline and intralaminar thalamus (TH) and basolateral nucleus of the amygdala (BLA). It is unclear which if any of these afferents innervate the medium spiny neurons of the NAc, that project to the VP. To identify the source of glutamatergic afferents that innervate neurons projecting to the VP, a dual-labelling method was used: Phaseolus vulgaris leucoagglutinin for anterograde and EGFP-encoded adenovirus for retrograde tract-tracing. Within the NAc, anterogradely labelled BLA terminals formed asymmetric synapses on dendritic spines that belonged to medium spiny neurons retrogradely labelled from the VP. TH terminals also formed synapses on dendritic spines of NAc neurons projecting to the VP. However, dendrites and dendritic spines retrogradely labelled from VP received no direct synaptic contacts from afferents originating from mPFC and vHC in the present material, despite the large number of fibres labelled by the anterograde tracer injections. These findings represent the first experimental evidence for a selective glutamatergic innervation of NAc neurons projecting to the VP. The glutamatergic inputs of different origin (i.e. mPFC, vHC, BLA, TH) to the NAc might thus convey different types of reward-related information during goal directed behaviour, and thereby contribute to the complex regulation of nucleus accumbens functions. PMID- 21643648 TI - [Increase of entomological indices during the pre-epidemic period of dengue in Ben Tre, South Vietnam]. AB - Dengue has emerged in Vietnam 50 years ago and since has become endemo-epidemic throughout the whole country. Each year, major epidemics of dengue fever (DF) and dengue hemorrhagic fever (DHF) hit South Vietnam during the rainy season, causing significant morbidity and mortality, especially among young children. The only preventive measure is vector control, but it is often implemented too late or indiscriminately. The aim of this study was to investigate, in the pre-epidemic stage, the existence of significant changes in vector indices, which will predict DF/DHF outbreaks. We conducted a descriptive transversal study, repeated once a month for four months (March to June) in the village of Locthuan (province Ben Tre) in the Mekong's delta. Adult mosquitoes were caught in 30 houses, and larvae were collected in water holding containers of 50 houses. The houses were randomly selected. Vector densities were calculated according to the indices recommended by WHO. Virological analysis was carried out on lots of female Aedes and larvae in order to determine viral infection rates. Catches of adult mosquitoes collected 496 specimens including 329 Aedes, 139 Culex and 28 Anopheles. Aedes aegypti was present in 63% of visited homes that is an average density of 1.8 mosquitoes per house. The increase in imaginal indices during the 4 months was not significant. The survey of breeding sites of Ae. aegypti identified 1292 water containers in which 71,569 larval specimens were collected. The values of house index, container index [CI] and Breteau index [BI] increased each month, the latter from 166 to 442. This increase was significant for CI and BI. Breeding sites were mostly intra-home, mainly consisting of large and small ceramic jars. Larval density of Ae. aegypti in the containers also increased significantly over the 4 months. It was correlated with the lack of cover and predators such as Mesocyclops spp., Micronecta spp. and larvivorous fishes. Cultivation of 15 pools of 10 adult females and 29 pools of larvae (ie 1088 specimens) of Ae. aegypti failed to isolate dengue virus. The high Stegomyia indices measured in this South Vietnamese village and their increase before the rainy season reflect a situation at high risk of epidemics but cannot predict the occurrence of an outbreak in the absence of virus isolation from mosquitoes. They justify conducting an integrated vector control throughout the year. PMID- 21643649 TI - Ferritin and iron levels in children with autistic disorder. AB - Iron has an important role on cognitive, behavioral, and motor development. High prevalence of iron deficiency has been reported in autism. The aim of this study was to investigate iron status in a group of children with autistic disorder. The sample was composed of 116 children between 3 and 16 years with a diagnosis of autistic disorder according to DSM-IV criteria. Serum ferritin, iron, hemoglobin, hematocrit, mean corpuscular volume, and red cell distribution width values were measured. We found that 24.1% of subjects had iron deficiency, and 15.5% had anemia. There was a significant positive correlation between age and ferritin and hematological measures. Results of this study confirmed that iron deficiency and anemia are common in children with autistic disorder. CONCLUSION: These findings suggest that ferritin levels should be measured in subjects with autism as a part of routine investigation. PMID- 21643650 TI - Usefulness of wireless capsule endoscopy for detecting inflammatory bowel disease in children presenting with arthropathy. AB - Inflammatory bowel disease (IBD) is a cause of chronic intestinal inflammation in children. In a subset of patients affected by IBD, arthropathy may be the leading presenting sign. In the past years, remarkable advances in gastrointestinal endoscopy techniques have been achieved; recently, the development of capsule endoscopy (CE) provided a non-invasive method for the complete endoscopic evaluation, including small bowel assessment. We report three children suffering from IBD but presenting with articular complaints in whom CE was a useful tool for detecting gut inflammation. Patients were investigated with the wireless CE: PillCam SB2 (Given Imaging, Yoqneam, Israel) capsule, the second-generation capsule, was used in our paediatric patients. Three patients were initially evaluated for arthropathy. Enteropathic arthritis was suspected for gastrointestinal symptoms and/or persistence of inflammatory markers elevation. In one of these children, conventional endoscopy was refused by parents, while in the other two children, CE was proposed as first-line diagnostic tool. In all patients, CE revealed to be safe and provided information that led to diagnosis. Paediatric rheumatologists should consider CE as a valid, non-invasive tool, eventually first level diagnostic approach in order to evaluate the presence of IBD in children presenting with chronic articular complaints. PMID- 21643651 TI - Familial hypocalciuric hypercalcemia: new mutation in the CASR gene converting valine 697 to methionine. AB - Familial hypocalciuric hypercalcemia is an uncommon cause of hypercalcemia that arises from mutations in the calcium-sensing receptor gene. Inactivation of this receptor leads to a decreased receptor sensitivity to calcium, determining that higher concentrations of calcium are needed to inhibit the release of parathormone in the parathyroid glands. Patients usually are asymptomatic. Diagnosis is usually made casually after a routine blood analysis. The syndrome is characterized by mild or moderate hypercalcemia, hypocalciuria, and normal or slightly increased levels of parathormone. The degree of hypercalcemia depends on the type of mutation. The accurate diagnosis is important since it is a benign disorder that does not require medical or surgical treatment. We report a 9-year old female with persistent hypercalcemia in several routine blood analyses, who was diagnosed with familial hypocalciuric hypercalcemia after genetic studies were performed. A new mutation determining a nucleotide change c.2089G>A in the calcium-sensing receptor gene (exon 7) was detected. This mutation was also found in the patient's mother and brother. PMID- 21643652 TI - Nocturnal apnea in Chiari type I malformation. AB - A 4-year-old girl presented with sleep-disordered breathing. Her parents described breathing pauses of up to 20 s and progressive tiredness during the day. Obstructive apneas from an enlarged adenoid were thought to be the most probable cause. However, an adenotomy did not resolve the problem. Polysomnography demonstrated central apneas, and cerebral magnetic resonance imaging revealed a Chiari type I malformation. We describe the differential diagnosis of apnea in children and the role of polysomnography in the distinction between obstructive and central apneas. CONCLUSION: This case illustrates that, in children with apnea, it is important to consider central causes as well as the more common obstructive causes, even in the absence of additional neurological signs or symptoms. PMID- 21643653 TI - Glucose-dependent docking and SNARE protein-mediated exocytosis in mouse pancreatic alpha-cell. AB - The function of alpha-cells in patients with type 2 diabetes is often disturbed; glucagon secretion is increased at hyperglycaemia, yet fails to respond to hypoglycaemia. A crucial mechanism behind the fine-tuned release of glucagon relies in the exocytotic machinery including SNARE proteins. Here, we aimed to investigate the temporal role of syntaxin 1A and SNAP-25 in mouse alpha-cell exocytosis. First, we used confocal imaging to investigate glucose dependency in the localisation of SNAP-25 and syntaxin 1A. SNAP-25 was mainly distributed in the plasma membrane at 2.8 mM glucose, whereas the syntaxin 1A distribution in the plasma membrane, as compared to the cytosolic fraction, was highest at 8.3 mM glucose. Furthermore, following inclusion of an antibody against SNAP-25 or syntaxin 1A, exocytosis evoked by a train of ten depolarisations and measured as an increase in membrane capacitance was reduced by ~50%. Closer inspection revealed a reduction in the refilling of granules from the reserve pool (RP), but also showed a decreased size of the readily releasable pool (RRP) by ~45%. Disparate from the situation in pancreatic beta-cells, the voltage-dependent Ca2+ current was not reduced, but the Ca2+ sensitivity of exocytosis decreased by the antibody against syntaxin 1A. Finally, ultrastructural analysis revealed that the number of docked granules was >2-fold higher at 16.7 mM than at 1 mM glucose. We conclude that syntaxin 1A and SNAP-25 are necessary for alpha-cell exocytosis and regulate fusion of granules belonging to both the RRP and RP without affecting the Ca2+ current. PMID- 21643654 TI - Inducible expression of TGFbeta, snail and Zeb1 recapitulates EMT in vitro and in vivo in a NSCLC model. AB - The progression of cancer from non-metastatic to metastatic is the critical transition in the course of the disease. The epithelial to mesenchymal transition (EMT) is a mechanism by which tumor cells acquire characteristics that improve metastatic efficiency. Targeting EMT processes in patients is therefore a potential strategy to block the transition to metastatic cancer and improve patient outcome. To develop models of EMT applicable to in vitro and in vivo settings, we engineered NCI-H358 non-small cell lung carcinoma cells to inducibly express three well-established drivers of EMT: activated transforming growth factor beta (aTGFbeta), Snail or Zeb1. We characterized the morphological, molecular and phenotypic changes induced by each of the drivers and compared the different end-states of EMT between the models. Both in vitro and in vivo, induction of the transgenes Snail and Zeb1 resulted in downregulation of epithelial markers and upregulation of mesenchymal markers, and reduced the ability of the cells to proliferate. Induced autocrine expression of aTGFbeta caused marker and phenotypic changes consistent with EMT, a modest effect on growth rate, and a shift to a more invasive phenotype. In vivo, this manifested as tumor cell infiltration of the surrounding mouse stromal tissue. Overall, Snail and Zeb1 were sufficient to induce EMT in the cells, but aTGFbeta induced a more complex EMT, in which changes in extracellular matrix remodeling components were pronounced. PMID- 21643655 TI - Antiplasmodial potential of medicinal plant extracts from Malaiyur and Javadhu hills of South India. AB - The emergence and spread of Plasmodium falciparum with resistance to chloroquine (CQ), the safest and cheapest anti-malarial drug, coupled with the increasing cost of alternative drugs especially in developing countries have necessitated the urgent need to tap the potential of plants for novel anti-malarials. The present study investigates the anti-malarial activity of the methanolic extracts of 13 medicinal plants from the Malaiyur and Javadhu hills of South India against blood stage CQ-sensitive (3D7) and CQ-resistant (INDO) strains of P. falciparum in culture using the fluorescence-based SYBR Green I assay. Sorbitol-synchronized parasites were incubated under normal culture conditions at 2% hematocrit and 1% parasitemia in the absence or presence of increasing concentrations of plant extracts. CQ and artemisinin were used as positive controls, while 0.4% DMSO was used as the negative control. The cytotoxic effects of extracts on host cells were assessed by functional assay using HeLa cells cultured in RPMI containing 10% fetal bovine serum, 0.21% sodium bicarbonate and 50 MUg/mL gentamycin (complete medium). Plant extracts (bark methanol extracts of Annona squamosa (IC(50), 30 MUg/mL), leaf extracts of Ocimum gratissimum (IC(50), 32 MUg/mL), Ocimum tenuiflorum (IC(50), 31 MUg/mL), Solanum torvum (IC(50), 31 MUg/mL) and Justicia procumbens (IC(50), 63 MUg/mL), showed moderate activity. The leaf extracts of Aristolochia indica (IC(50), 10 MUg/mL), Cassia auriculata (IC(50), 14 MUg/mL), Chrysanthemum indicum (IC(50), 20 MUg/mL) and Dolichos biflorus (IC(50), 20 MUg/mL) showed promising activity and low activity was observed in the flower methanol extracts of A. indica , leaf methanol extract of Catharanthus roseus, and Gymnema sylvestre (IC(50), >100 MUg/mL). These four extracts exhibited promising IC(50) (MUg/mL) of 17, 24, 19 and 24 respectively also against the CQ resistant INDO strain of P. falciparum. The high TC(50) in mammalian cell cytotoxicity assay and the low IC(50) in anti-malarial P. falciparum assay indicates selectivity and good resistance indices in the range of 0.9-1.7 for leaf extracts of A. indica, C. auriculata, C. indicum and D. biflorus suggests that these may serve as anti-malarial agents even in their crude form. These results indicate a possible explanation of the traditional use of some of these medicinal plants against malaria or malaria-like conditions. PMID- 21643656 TI - Comparison of dose at an interventional reference point between the displayed estimated value and measured value. AB - Today, interventional radiology (IR) X-ray units are required for display of doses at an interventional reference point (IRP) for the operator (IR physician). The dose displayed at the IRP (the reference dose) of an X-ray unit has been reported to be helpful for characterizing patient exposure in real time. However, no detailed report has evaluated the accuracy of the reference doses displayed on X-ray equipment. Thus, in this study, we compared the displayed reference dose to the actual measured value in many IR X-ray systems. Although the displayed reference doses of many IR X-ray systems agreed with the measured actual values within approximately 15%, the doses of a few IR units were not close. Furthermore, some X-ray units made in Japan displayed reference doses quite different from the actual measured value, probably because the reference point of these units differs from the International Electrotechnical Commission standard. Thus, IR physicians should pay attention to the location of the IRP of the displayed reference dose in Japan. Furthermore, physicians should be aware of the accuracy of the displayed reference dose of the X-ray system that they use for IR. Thus, regular checks of the displayed reference dose of the X-ray system are important. PMID- 21643657 TI - Triterpenes involved in the anti-inflammatory effect of ethanolic extract of Pterodon emarginatus Vogel stem bark. AB - Bioassay-guided fractionation of the ethanolic extract of Pterodon emarginatus Vogel stem bark (EtEx) resulted in the isolation and characterization of lupeol and betulin. Their structures were elucidated by spectroscopic methods including IR, (1)H-NMR, (13)C-NMR and comparison with literature values. This study showed the anti-inflammatory activity of EtEx, the hexane (HexL) and dichloromethane (DichL) layers, and lupeol and betulin. The extract, HexL, DichL, lupeol and betulin were able to inhibit acetic acid-induced writhing. In the formalin test, EtEx decreased licking time only in the second phase characterizing anti inflammatory activity. In the oil-induced ear oedema test, EtEx, lupeol and betulin decrease edema formation. In conclusion, EtEx has antinociceptive effects arising from anti-inflammatory activity; this activity could be due to the presence of lupeol and betulin. PMID- 21643659 TI - Pathology and molecular diagnosis of paratuberculosis of camels. AB - Camels are the prime source of meat and milk in many desert regions of the world including Saudi Arabia. Paratuberculosis of camels, locally called Silag, is a serious and invariably fatal disease in the Arabian camel. Six camels were used in this study. Five camels with clinical paratuberculosis were used to study the pathology of the disease and confirm its aetiology. The sixth camel was clinically healthy and used as a control. The camels were examined clinically and bled for haematological and blood chemistry analysis. They were then humanely killed with a high intravenous dose of thiopental sodium (10 mg/kg) for pathological studies as well as obtaining tissues for microbiological and molecular studies. The clinical signs of the disease were emaciation, diarrhoea, alopecia, wry neck and pale mucous membranes. Laboratory diagnosis showed reduced haemoglobin concentration, low haematocrit and high activity of the serum enzyme alanine aminotransferase. Serum creatinine concentration was normal. These results indicated the infected camels were anaemic and the function of their livers was affected. Postmortem examination showed thickened and corrugated intestinal mucosa, enlarged granulomatous mesenteric lymph nodes, miliary and diffuse granulomas in the liver (in four camels), generalized lymph node granulomas (in one camel), splenic granuloma (in one camel) and mediastinal lymph node granuloma (in two camels). Histopathological examination showed diffuse infiltration of macrophages in all organs showing lesions. Ziehl-Neelsen staining of tissue scraping and tissue sections showed masses of acid fast bacilli, except for the spleen. Infection with Mycobacterium avium subsp. paratuberculosis was confirmed by PCR by targeting the IS900 gene. PMID- 21643660 TI - Anthelmintic screening of fractions of Elephantorrhiza elephantina root extract against Haemonchus contortus. AB - In the previous work conducted, it was found that the root extract of Elephantorrhiza elephantina (Bruch.) Skeels. exhibited good anthelmintic efficacy against eggs and larvae of Haemonchus contortus. This study was therefore undertaken to screen fractions of the extracts of E. elephantina for their anthelmintic activity against adult H. contortus using a bioactivity-guided assay. The adult worm bioassay was conducted using aqueous, hexane and ethyl acetate fractions of the plant extracts at concentrations of 0.312, 0.625, 1.25, 2.5, 5 and 10 mg/ml in vitro. Albendazole and distilled water were used as positive and negative control, respectively. In vitro treatment of adult worms revealed concentration and time-dependent efficacy of the fractions of E. elephantina. The aqueous and ethyl acetate fractions showed a highly significant (P<0.05) motility inhibition at concentrations of 2.5 mg/ml and above after 6 h of exposure, while the hexane fraction showed a significant (P<0.05) motility inhibition at concentrations of 5 mg/ml and above. After 30 h of exposure, all the fractions showed insignificant (P>0.05) inhibition of motility. The fractions of E. elephantina also showed mortality indexes that were not significantly different to each other, as well as to the commercial drug (albendazole). Overall, the ethyl acetate and the aqueous fractions were found to possess the highest anthelmintic activity. We therefore recommend future studies to further investigate effective dosages of these fractions in vivo, as well as a full investigation into the compounds responsible for the biological activity. PMID- 21643661 TI - Follicular characteristics and intrafollicular concentrations of nitric oxide and ascorbic acid during ovarian acyclicity in water buffalo (Bubalus bubalis). AB - The objective of this study was to examine the follicular characteristics and intrafollicular concentrations of nitric oxide and ascorbic acid during ovarian acyclicity in buffaloes. Ovaries were collected from 56 acyclic and 95 cyclic buffaloes at slaughter, surface follicle number was counted and follicles were classified into small (5.0-6.9 mm), medium (7.0-9.9 mm), and large (>= 10.0 mm) size categories based on their diameter. Follicular fluid was aspirated and assayed for nitric oxide, ascorbic acid, estradiol, and progesterone. Acyclic buffaloes had a higher (P<0.05) number of medium-sized follicles and a lower (P<0.001) number of large follicles than the cyclic ones. In acyclic animals, the number of large follicles was lower (P<0.01) than in medium size category which in turn was lower (P<0.001) than the number of small follicles. In contrast, the number of medium and large follicles was not different (P>0.05) in the cyclic control. However, the number of small-sized follicles was higher (P<0.001) compared to the other two categories. The incidence of large-sized follicles was lower (P<0.05) in acyclic buffalo population compared to the cyclic control. Evaluation of estrogenic status demonstrated that all the follicles of acyclic buffaloes are estrogen-inactive (E (2)/P (4) ratio<1). Small- and medium-sized follicles of acyclic buffaloes had higher concentrations of nitric oxide (P<0.05 and P<0.001, respectively) and lower concentrations of ascorbic acid (P<0.05 and P<0.01, respectively) than the corresponding size estrogen-active follicles of their cyclic counterparts. In conclusion, this study indicates that follicular development continues during acyclicity in buffaloes. Although follicles in some acyclic buffaloes attain a size corresponding to morphological dominance, they are unable to achieve functional dominance, perhaps due to an altered balance of intrafollicular nitric oxide and ascorbic acid and, as a result, these follicles instead of progressing to ovulation undergo atresia. PMID- 21643662 TI - Maedi in slaughtered sheep: a pathology and polymerase chain reaction study in southwestern Iran. AB - Maedi-visna (MV) is an important slow viral disease of sheep leading to a progressive lymphoproliferative disease. It affects multiple organs primarily the lungs, where it causes interstitial pneumonia (maedi). In this study, the lungs of 1,000 sheep carcasses were grossly inspected and those suspected to have maedi were studied at histopathological and molecular levels. A polymerase chain reaction (PCR) technique that amplified a 291-base pair DNA in the long terminal repeat (LTR) sequence of MV provirus was conducted on all the 50 suspected lungs together with 10 normal appearing lungs as controls. Amplicons of the expected size were detected in 11 (n=11/50) suspected sheep, and one of the 10 control sheep. Histopathologic study of the pulmonary lesions of all 11 (n=11/11) positive sheep showed MV lesions, including hyperplasia of the perivascular and peribronchiolar lymphoid cells, interstitial lymphoplasmacytic infiltration and smooth muscle hyperplasia and the histopathologic findings were correlated with PCR results. In contrast, the tissue sections of control animals were almost normal at histopathological level; however, PCR technique demonstrated that one of them was affected by maedi. This study showed that the LTR-PCR had high specificity and sensitivity in diagnosis of this viral infection. This study is the first to evaluate the prevalence of MV virus infection in sheep in Iran. PMID- 21643663 TI - Gastrointestinal and blood parasite determination in the guanaco (Lama guanicoe) under semi-captivity conditions. AB - The breeding of wild animals for commercial purposes is becoming more frequent nowadays. This situation has led to an increase in contact rates between wild and domestic animals, with subsequent reciprocal transmission of parasites. In this study, we characterized the gastrointestinal and blood parasites of a group of 15 semi-captive guanacos (Lama guanicoe). We characterized gastrointestinal parasites by analyzing fecal samples through the sedimentation-flotation technique and hemoparasites by using blood smears stained with Giemsa. We found several gastrointestinal parasites including Nematoda and protozoans. The most frequently found parasites were Nematodirus sp. and Eimeria sp. In contrast with previous studies, neither Cestoda nor Fasciola were found. The only hemoparasite detected was Mycoplasma haemolamae, a parasite already described in llamas and alpacas. We conclude that the most frequent gastrointestinal parasites of semi captive guanacos were nematodes and protozoans. Also, the hemoparasite M. haemolamae seems to be prevalent among captive populations of South American camelids. Finally, captive guanacos share several parasites with the traditional livestock. Therefore, keeping captive or semi-captive guanacos without an adequate sanitary protocol might have adverse consequences to adjacent traditional cattle farming and/or for wild animals. PMID- 21643664 TI - Survey of brucellosis at the wildlife-livestock interface on the Zimbabwean side of the Great Limpopo Transfrontier Conservation Area. AB - A cross-sectional study was conducted to determine the seroprevalence of bovine brucellosis in communal cattle and wildlife at a wildlife-livestock interface in the southeast lowveld of Zimbabwe, part of the Great Limpopo Transfrontier Conservation Area. RBT and c-Elisa were used in serial for detection of antibodies against Brucella spp. Between July 2007 and October 2009, a total of 1,158 cattle were tested and the overall seroprevalence of brucellosis was 9.9%. A total of 97 wild animals (African buffaloes (n=47), impala (n=33), kudu (n=16), and giraffe (n=1)) were tested and only one animal (giraffe) was seropositive for brucellosis (1.03%). Brucella seroprevalence showed an increasing trend with age, with adult cattle (>6 years) recording the highest seroprevalence (11.1%), but the differences were not statistically significant. Similarly, female cattle recorded a relatively higher seroprevalence (10.8%) compared to males (7.9%), but the difference was not significant. However, a significant (P<0.001) association between Brucella seropositivity and abortion history was recorded in female cattle. Similarly, Brucella seropositivity was significantly (P<0.01) associated with a history of grazing in the park for female cattle. Overall, from the interface area, cattle with a history of grazing in the park recorded a significantly (P<0.01) higher Brucella seroprevalence (13.5%) compared to those with no history of grazing in the park (4.9%). The significant association between abortion history and seropositivity observed in this study illustrates the potential economic significance of Brucella in cattle in this area. Hence, public awareness and further epidemiological studies of the disease in wildlife, livestock, and humans in the study area are of great importance. PMID- 21643665 TI - Smallholder dairy production in Northern Malawi: production practices and constraints. AB - Milk production in Malawi is still unsatisfactory despite efforts by different stakeholders to boost the dairy sector. To investigate the roots of the problem, a survey on the current production practices and constraints on smallholder dairy farming was conducted in the Northern Region of the country. A total of 210 farmers were interviewed. The results revealed that farmers had small herd sizes in the region with an average of 2.2 +/- 0.6 cattle per farmer. Average herd size was larger in male-managed farms than in female-managed farms (2.6 +/- 2.8 vs. 1.8 +/- 1.3), farmers with more than 5 years of dairy farming experience had larger herds than those with less experience (2.6 +/- 2.8 vs. 1.9 +/- 1. 2) and farmers who grazed their animals tended to have larger herds than those that stall-fed their animals (4.4 +/- 5.1 vs. 1.9 +/- 1.3). Average milk production was 8.2 +/- 6.5 l per cow per day. Higher average daily milk production was observed in farmers with above primary school education (10.3 +/- 8.3 vs. 7.7 +/- 5.6), those with dairy farming as main activity (9.3 +/- 6.6 vs. 6. 5 +/- 6.1) and farmers with more than 2 years of experience in dairy farming (9.3 +/- 6.3 vs. 6.1 +/- 6.4). Unreliable supply of improved animal genetics, poor animal health, feed shortage and poor prices for milk were considered to be the most important constraints to smallholder dairy farming in descending order. PMID- 21643666 TI - Ovine and caprine toxoplasmosis (Toxoplasma gondii) in aborted animals in Jordanian goat and sheep flocks. AB - Two hundred and fifty-five biological samples (106 aborted foetal tissue samples and 149 blood samples from aborted sheep and goats) were collected from 188 animals during the lambing season from September 2009 to April 2010 from the Mafraq region of Jordan. The sampled animals belonged to 93 goat and sheep flocks that had cases of abortion. A total of 169 (66.3%) biological samples were collected from sheep and 86 (33.7%) from goats. Seventy-six (29.8%) biological samples (45 blood and 31 tissue samples) were positive for Toxoplasma gondii by PCR assay. The positive samples were obtained from 43 sheep and 23 goats. The overall toxoplasma-specific prevalence rate was 35.1% (66/188). Forty flocks (43%) had at least one T. gondii PCR-positive animal. The risk factors related to flock health status and farm management that are hypothesized to be associated with T. gondii PCR positivity were also assessed using multiple logistic regressions. The presence of cats (OR = 4.74), a large flock size (OR = 2.76) and the method of disposing the aborted foetuses (OR = 3.77) were all statistically significant (P<0.05) risk factors that were positively associated with toxoplasma positivity in goat and sheep flocks. PMID- 21643667 TI - Psoroptes cuniculi induced oxidative imbalance in rabbits and its alleviation by using vitamins A, D3, E, and H as adjunctive remedial. AB - The oxidant/antioxidant balance of rabbits naturally infected with Psoroptes cuniculi and treated with ivermectin +/- vitamins A, D(3), E, and H supplementation was investigated. Two groups of seven mixed ? and ?, 6-to-8 month old New Zealand White rabbits, diagnosed Psoroptes mites-positive by skin scraping examination and seven clinically healthy control rabbits were examined. Blood samples were obtained on day 0 and at 28 days post-therapy to determine oxidative stress indices. On day 0, the levels of lipid peroxides were significantly higher (P <= 0.01) in the Psoroptes-infected rabbits compared with the healthy controls while those of reduced glutathione and the activities of the antioxidant enzymes glutathione peroxidase, glutathione-S-transferase, catalase, and superoxide dismutase were significantly lower (P <= 0.01). Vitamin supplementation of the ivermectin-treated rabbits revealed both faster clinical (14 days) and parasitological (10 days) recovery. It was concluded that significant alteration of oxidant/antioxidant balance is a factor in the pathogenesis of P. cuniculi infestation of rabbits, and recovery can be enhanced by combining ivermectin treatment with vitamin A, D(3,) E, and H supplementation. PMID- 21643668 TI - Detection of superantigenic toxin genes in Staphylococcus aureus strains from subclinical bovine mastitis. AB - The aim of this study was to determine the presence of genes encoding enterotoxins (sea-sej) and toxic shock syndrome toxin-1 (tst) of Staphylococcus aureus strains (n = 130) isolated from subclinical bovine mastitis in Turkey by polymerase chain reaction. Sixty-one (46.9%) isolates were found to contain one or more toxin genes. The most frequently found enterotoxin genes were seg (16.2%) and sei (16.2%), followed by sec (15.4%), sed (10.8%), and sej (10.8%), respectively. The tst gene was detected in seven (5.4%) isolates. None of S. aureus strains harbored sea, seb, see, and seh genes. Since these toxins are recognized agents of staphylococcal food poisoning, it must be considered that the consuming raw milk and raw milk products would pose public health risk as high prevalence of toxigenic S. aureus was found in this study. PMID- 21643669 TI - Is educational achievement a turning point for incarcerated delinquents across race and sex? AB - Research has linked the role of education to delinquency, but much of the focus has been on general population samples and with little attention to demographic differences. Employing a cumulative disadvantage framework that integrates elements of informal social control and labeling theories, this article examines whether academic achievement serves as a positive turning point and re-directs juvenile delinquents away from subsequent offending. Attention is also given to race/sex contingencies. Using a sample of 4,147 delinquents released from Florida correctional institutions (86% male, 57% non-White, average age at release = 16.8 years), propensity score analysis yielded two findings: youth with above average academic achievement while incarcerated were significantly more likely to return to school post-release, and youth with above average attendance in public school were significantly less likely to be re-arrested in the 1-year post-release period. While the academic gains were pronounced among African-American males, the preventive effects of school attendance are similar across race and sex, suggesting that education can be a part of a larger prevention effort that assists juvenile delinquents in successful community re-entry. PMID- 21643670 TI - Invited address: James Joyce, Alice in Wonderland, The Rolling Stones, and criminal careers. AB - The study of criminal careers generally, and patterns of continuity and change in criminal offending in particular, has been a long-standing interest to social scientists across many disciplines. This article provides readers with an overview of this line of research. After an introduction to the criminal career perspective, the article presents several 'facts' that have emerged from criminal career studies. This material segues into a discussion of theories based on criminal careers research as well as a related discussion of the emerging methods and trends in the area. The article closes with some observations about public policy with respect to criminal careers knowledge and identifies some neglected research needs. A key summary conclusion is that the processes associated with continuity and change are not mutually exclusive, but instead are important and complimentary aspects of criminal careers research. PMID- 21643671 TI - Clipping of MCA aneurysms: how I do it. AB - INTRODUCTION: Aneurysms located at the middle cerebral artery bifurcation remain a clear neurosurgical indication. We detail here the steps necessary to enable safe surgery for Sylvian fissure aneurysms. METHODS: An angiogram with 3D reconstruction is obtained and reviewed intraoperatively, just prior to the skin incision. During the exposure, the cistern is kept open by small cottonoids, thereby avoiding brain retraction. Continuous monitoring of MEPs along with ICG microscopic angio-fluorescence allows for detection of vascular compromise. Intraoperative angiography with 3D reconstruction allows for immediate correction of less than satisfactory surgical outcome. CONCLUSIONS: Careful planning of surgical strategy followed by a minimally invasive technique (with continuous neuro-monitoring) ensures safe surgery. The availability of intra-operative radiological guidance allows for optimal management. PMID- 21643672 TI - Intracranial clear cell meningioma: a clinicopathologic study of 15 cases. AB - OBJECT: Clear cell meningioma (CCM) is a rare histological variant of meningioma. CCM has a high recurrence rate and aggressiveness. In this study, we reviewed our experience in the treatment of the lesion. METHODS: Here we present a series of 15 patients with intracranial CCM. The clinical data were retrieved from the records of our Neurosurgery Department and the patients' prognoses were attained by clinic service and telephone. Immunohistochemistry for epithelial membrane antigen (EMA), vimentin, glial fibrillary acidic protein (GFAP), CD10, and S-100 was done, and the MIB-1 labeling index was calculated in all cases. RESULTS: The 15 patients included eight males and seven females; the mean age was 34.8 years. The most frequent initial symptoms were headache and hearing loss. The most common location was the cerebellopontine angle (CPA) zone. Eleven patients had total removal and four patients underwent subtotal removal. Histological features of atypia were present in different proportions, from 6.7% to 100%, and six cases accorded with atypia. Three tumors showed brain invasion. EMA and vimentin were 100% positive, and CD10 was 100% negative. GFAP was 87% negative and S-100 was 93% negative. The mean follow-up period was 36.7 months. Three patients with brain invasion all recurred and five cases with atypia recurred. In 11 patients with total removal, six patients recurred. In four patients with subtotal removal, three patients recurred. Kaplan-Maier analysis showed that incomplete surgical resection was significantly associated with recurrence (p = 0.001). The MIB-1 labeling index for recurrence was 5.7 +/- 2.7% versus 2.8 +/- 1.5% for no recurrence (p = 0.036). CONCLUSIONS: CCM is a rare subtype of meningioma, with a tendency to present in younger patients and a propensity to recur. Immunohistochemistry plays a vital role in differentiating CCM from other tumors. Brain invasion, atypia and MIB-1 labeling index are likely to predict the recurrence. The extent of resection might be connected with the prognosis. PMID- 21643673 TI - Effects of antipsychotic treatment on psychopathology and motor symptoms. A placebo-controlled study in healthy volunteers. AB - RATIONALE: There is increased interest in elucidating the range of symptoms of schizophrenia and their response to treatment with medications. Particularly negative and cognitive symptoms are often resistant to the therapy with currently available antipsychotics. There are even similarities between negative symptoms in psychosis and the side effects of antidopaminergic antipsychotic drugs. OBJECTIVES: The aim of this randomized, single-blinded, placebo-controlled study was to investigate the influence of a subchronic, prolonged neuroleptic-induced dopamine deficit on psychopathology and subjective well-being in healthy subjects. METHODS: Seventy-two healthy volunteers without history of psychiatric diseases were included. A 7-day antidopaminergic intervention was provided with aripiprazole, haloperidol, and reserpine. For the clinical assessment, structured interviews and psychopathology and extrapyramidal symptom scales were used. RESULTS: Seven out of 18 participants (38.9%) randomized to the haloperidol group terminated the study ahead of schedule. In the reserpine and the haloperidol group, significantly higher levels of negative and positive symptoms (PANSS scale) were documented. Depressive symptoms predominantly occurred in the reserpine group. Among all participants experiencing the antidopaminergic intervention, the subgroup with positive family history among first and second generation relatives developed more pronounced depressive symptoms. Concerning extrapyramidal motor symptoms, the haloperidol group had significantly more severe manifestations than all three other groups. CONCLUSION: Antidopaminergic modulation in healthy subjects induced substantial impairments in several domains of subjective well-being. In particular an association between hypodopaminergic states and depressive symptoms was observed which may be amplified by a genetic predisposition. PMID- 21643674 TI - Work aversion and associated changes in dopamine and serotonin transporter after methamphetamine exposure in rats. AB - RATIONALE: Methamphetamine (mAMPH) administration in animals can lead to a variety of cognitive and behavioral deficits. We previously reported non-acute reversal learning impairments after a single-day administration of mAMPH, providing evidence of this drug's selective effects on inhibitory control. Effortful decision-making (i.e., how much effort to invest in rewards) is an aspect of cognition that has not yet been explored after mAMPH. OBJECTIVES: Given that frontostriatal circuitry mediating this type of choice is vulnerable to the effects of mAMPH, we tested the hypothesis that mAMPH may also affect decision making involving effort, another form of cognitive flexibility. METHODS: We examined the non-acute effects of an experimenter-administered single day of mAMPH on effort discounting. In this task, rats previously treated with mAMPH or saline (SAL) could select a high reward at the cost of climbing over a tall barrier or a low reward with no barrier impeding its procurement. RESULTS: Following treatment, mAMPH rats were more work-averse than SAL rats. A control task showed there were no treatment group differences when the high and low rewards involved equal work: all rats chose the high reward preferentially. There were no significant treatment group differences in [(125)I]RTI-55 binding to dopamine and serotonin transporters (DAT, SERT) in any of the regions assayed; however, there were significant correlations of accumbens DAT and cingulate SERT with post-treatment performance. CONCLUSIONS: These findings suggest that even modest dose mAMPH exposure has long-lasting effects on effortful decision-making and may do so through influences on forebrain monoaminergic systems. PMID- 21643675 TI - Effects of a stressor and corticotrophin releasing factor on ethanol deprivation induced ethanol intake and anxiety-like behavior in alcohol-preferring P rats. AB - RATIONALE: Stress may elevate ethanol drinking and anxiety associated with ethanol drinking. Studies to identify relevant neurobiological substrates are needed. OBJECTIVE: To assess roles of brain regions in corticotrophin releasing factor (CRF) effects on stressor-enhanced, ethanol deprivation-induced drinking and anxiety-like behavior. METHODS: Ethanol-preferring rats (P rats) were exposed to three cycles of a two-bottle choice paradigm with two 2-day deprivation periods that included 1 h exposure to a restraint stressor. To assess the role of CRF and to identify relevant brain regions, a CRF-1 receptor antagonist (SSR125543; 10 ug) was injected into the nucleus accumbens (NAC), amygdala (Amyg), or dorsal raphe nucleus (DRN) prior to exposure to the restraint stressor. In a second study, CRF (0.5 ug) was injected into one of these regions, or the ventral tegmental area (VTA), or paraventricular nucleus of the hypothalamus (PVN). RESULTS: Applying the restraint stressor during deprivation increased voluntary intake and sensitized anxiety-like behavior. Antagonist injection into the NAC prevented increased drinking without affecting anxiety like behavior, whereas injection into the Amyg or DRN prevented the anxiety-like behavior without affecting drinking. To confirm CRF actions in the stressor effect, CRF was injected into selected brain regions. NAC injections (but not the VTA, Amyg, DRN, or PVN) facilitated drinking but did not change anxiety-like behavior. Injections into the DRN or Amyg (but not PVN or VTA) enhanced anxiety like behavior. CONCLUSIONS: Results emphasize that a restraint stressor elevates ethanol intake and sensitizes ethanol deprivation-induced anxiety-like behavior through CRF1 receptors in the NAC and Amyg/DRN, respectively. PMID- 21643676 TI - Preclinical assessment of an adjunctive treatment approach for cognitive impairment associated with schizophrenia using the alpha7 nicotinic acetylcholine receptor agonist WYE-103914/SEN34625. AB - RATIONALE: alpha7 nicotinic acetylcholine receptor (nAChR) agonists are proposed as candidate agents for the adjunctive treatment of cognitive deficits associated with schizophrenia. Despite the pursuit of such an approach clinically, it is surprising that the preclinical profile of pro-cognitive agents in conjunction with antipsychotic drugs is currently unexplored. OBJECTIVES: We determined if the memory-enhancing effects of the selective alpha7 nAChR agonist WYE-103914 were preserved in the presence of the atypical antipsychotic drug risperidone, and if the antipsychotic-like profile of risperidone was preserved in the presence of WYE-103914. METHODS: Using the rat novel object recognition (NOR) paradigm, the maintenance of memory-enhancing activity of the alpha7 nAChR agonist WYE-103914 in the presence of risperidone was examined. Similarly, in the standard tests of antipsychotic-like activity, apomorphine-induced climbing (AIC) in mice and conditioned avoidance responding (CAR) in rats, the preservation of antipsychotic-like activity of risperidone was evaluated in the presence of WYE 103914. RESULTS: WYE-103914 exhibited memory-enhancing activity in rat NOR, and this effect of WYE-103914 was retained in the presence of risperidone. In AIC, the atypical antipsychotic profile of risperidone was not significantly altered by WYE-103914. In contrast, WYE-103914 moderately potentiated the efficacy profile of risperidone in CAR, an effect that did not appear to be convincingly linked to a pharmacokinetic interaction. CONCLUSIONS: These data underscore the value of a preclinical evaluation of the adjunctive profile of a memory-enhancing agent in combination with antipsychotics and provide further support to augmentation with alpha7 nAChR agonists to address the cognitive deficits associated with schizophrenia. PMID- 21643677 TI - Nicotine preloading: the importance of a pre-cessation reduction in smoking behavior. PMID- 21643678 TI - AL amyloidosis with severe gastrointestinal invasion and acute obstructive suppurative cholangitis. PMID- 21643679 TI - Successful management with an effective induction regimen followed by allogeneic hematopoietic stem cell transplantation for promyelocytic blast crisis of chronic myelogenous leukemia. PMID- 21643680 TI - von Willebrand disease: a clinical and laboratory study of sixty-five patients. AB - von Willebrand disease (VWD) is the commonest inherited bleeding disorder, yet it has not been well recognized in Southeast Asia. The aim of this prospective study was to report our experience of VWD diagnosis and to establish the clinical presentations of VWD in Taiwan. From October 2003 to April 2010, 863 patients with suspicion of having an inherited bleeding disorder underwent VWD screening tests. Those with positive tests were selected for further clinical and laboratory evaluation. A nested gender- and age-matched control cohort underwent similar investigation for comparison. VWD was diagnosed by comprehensive laboratory tests including factor VIII clotting activity, von Willebrand factor antigen assay, VWF:ristocetin cofactor activity (VWF:RCo) and platelet function analyzer (PFA)-100 closure times. VWF multimer analysis was performed by western blot for disease subtype identification. Sixty-five (7.5%) patients from 55 unrelated families were discovered to have VWD. Their median age was 27 years with a range of 4 to 69 years. The most common and specific bleeding symptom in male and female patients was bleeding after dental extraction and menorrhagia, respectively, as compared with control subjects. PFA-100 epinephrine closure time was the most sensitive laboratory test for VWD diagnosis with a sensitivity of 85%, followed by VWF:RCo assay (73%). Among 49 patients with VWF multimer analysis, 37(75.5%) were revealed to have type 1 VWD. Our study demonstrates that VWD and its clinical manifestations and subtypes in Taiwan are similar to those in the West and represents the first report of its kind in a Southeast Asian population. PMID- 21643681 TI - Long-term follow-up survey reveals a high yield, up to 30% of patients presenting newly detected aneurysms more than 10 years after ruptured intracranial aneurysms clipping. AB - The need to pursue long-term follow-up in patients treated for a ruptured aneurysm remains debated. New aneurysms development is a crucial element to consider but remains scarcely analyzed especially after a mean follow-up longer than 10 years. Our study was designed to provide rates of newly developed aneurysms in patients who have undergone prior clipping who were not followed with serial imaging. Patients were included if they were (1) treated more than 10 years ago by clipping of a ruptured aneurysm, (2) independent at time of discharge, (3) presently younger than 65 years, and if (4) they agreed to undergo a late digital subtraction angiography (DSA) control or to transmit results of a recent one performed elsewhere. Twenty patients were included with a mean delay between aneurysm treatment and late DSA of 18.0 years (10-26.5 years). Out of these patients, six (30%) harbored new aneurysms. Of these six individuals, four (66.6%) presented multiple aneurysms with a total of 15 newly discovered aneurysms. Aneurysm sizes ranged from 1 to 10 mm. One patient suffered from a de novo aneurysm rupture. Multiple aneurysms at the time of the first hemorrhage were a risk factor in developing de novo aneurysm (p=0.0175). In conclusion, based on a 30% rate of new aneurysm formation in patients clipped more than a decade ago, close screening on a very long-term perspective is encouraged. This study suggests aneurysm formation to be a continuous process. PMID- 21643682 TI - Usefulness of intraoperative laser Doppler flowmetry and thermography to predict a risk of postoperative hyperperfusion after superficial temporal artery-middle cerebral artery bypass for moyamoya disease. AB - We investigated whether postoperative hyperperfusion in moyamoya disease can be predicted using intraoperative laser Doppler flowmetry and/or thermography. A prospective study was conducted on 27 patients (39 hemispheres) with moyamoya disease who underwent superficial temporal artery-middle cerebral artery (STA MCA) bypass. During surgery, regional cerebral blood flow (rCBF) was measured with a laser Doppler flowmeter and the temperature of the cortical surface was measured with an infrared thermograph. Postoperative hyperperfusion was assessed immediately after surgery based on CBF study under sedation (propofol) as >100% increase in corrected rCBF compared to preoperative values. Postoperative hyperperfusion on CBF was observed in two patients (7.4%). A significant correlation was observed between intraoperative rCBF changes and postoperative rCBF increase (Pearson's method: r=0.555, p=0.0003; simple regression: Y=1.22X+3.289, r (2)=0.308, p=0.0004). Furthermore, the rCBF changes measured by laser Doppler flowmetry were significantly greater in patients with postoperative hyperperfusion (p=0.0193) and CHS (p=0.0193). The present study suggests that intraoperative rCBF measurement using laser Doppler flowmetry may predict a risk of post-EC-IC bypass cerebral hyperperfusion in moyamoya disease. PMID- 21643683 TI - Percutaneous transluminal angioplasty for atherosclerotic stenosis of the subclavian or innominate artery: angiographic and clinical outcomes in 36 patients. AB - The purpose of the study was to evaluate stenting and percutaneous transluminal angioplasty (PTA) for the treatment of stenotic lesions of the subclavian or innominate artery based on surgical results and long-term follow-up with 36 patients. In particular, we evaluated the efficacy of self-expanding stents compared to balloon-expandable stents. Between February 2000 and March 2008 at the Kyoto Medical Center, 36 patients underwent both stenting and PTA of the subclavian or innominate artery. Twenty-four patients had severe subclavian stenotic disease, ten patients had total occlusion of the subclavian artery, and two patients had stenoses of the innominate artery. Successful dilatation (less than 30% residual stenosis) was obtained in 34 of the 36 cases. In two cases (20%) of total subclavian occlusion, the guidewires were not able to penetrate the lesions, although the success rate was 100% for stenoses. All patients had no signs of neurological side effects with the exception of two pseudoaneurysms of the femoral arteries that required surgical intervention. In the first 30 days after treatment, there were no strokes or deaths. Outpatient follow-up was done with 30 patients (83.3%) after a mean of 30.9 months (range 3-114). Among these 30 patients, four patients (13.3%) developed restenoses of over 50%. Restenoses occurred in 4 of 20 individuals (20%) who received balloon-expandable stents but were not observed in those who received self-expanding stents. Endovascular therapy for the subclavian and innominate arteries is less invasive and safer than open surgery, making it the preferable option. In this clinical period, the rate of restenosis using self-expanding stents was lower than the rate using balloon-expandable stents. PMID- 21643684 TI - Assessment of waters and sediments impacted by drainage at the Young Dong coal mine site, South Korea. AB - INTRODUCTION: This study focused on the assessment of the geochemistry and hydrology of the Imgok Creek-Young Dong tributary for the design of a field coal mine drainage treatment system. METHODS AND RESULTS: Examination of this site showed that the pH was greatly lowered by the addition of the Young Dong water, except in the month of March. The alkalinity was also affected; the concentrations of iron, aluminum, and sulfate were elevated at sites below the confluence; of these, iron was particularly problematic. High iron concentrations were primarily restricted to the acid rock drainage (ARD) (YD-9) water sources, whereas high aluminum concentrations were seen in both the ARD and in some of the upstream water sources. The acidity was primarily due to ferrous and ferric iron with a lesser amount of aluminum acidity. Except for the sampling in March, the flow was dominated by the ARD. This hydrologic condition resulted from the loads of iron, aluminum, sulfate, and acidity, among other constituents, that were dominated by the ARD. CONCLUSION: Finally, treatment activities should primarily focus on the ARD and specifically seek to remove ferrous and ferric iron from the treatment system. PMID- 21643685 TI - Single stage corpectomy and instrumentation in the treatment of pathological fractures in the lumbar spine. AB - PURPOSE: Corpectomy and implantation of titanium cages is standard in pathological fracture treatment but additional single ventral instrumentation remains controversial with regard to rotational stability. METHODS: This study included 45 patients suffering from vertebral metastases with spinal stenosis, instability and/or neurological deficits secondary to pathological lumbar spine fractures and bone mineral density (BMD) >= 1.20 g/cm(2). The clinical results of a single stage anterior decompression with corpectomy defect restoration with titanium cage and single double rod system in patients were evaluated at mean 36 months postoperatively with follow-up neurological and radiological exams at three months then every six months. Evaluation of neurological recovery included grading following a modified Frankel scale. Contentment, disability and actual pain were evaluated using the visual analogue scale (VAS) and Oswestry disability index (ODI). BMD was measured using dual-energy X-ray absorptiometry (DXA). RESULTS: Postoperative neurological evaluations showed improvement in all patients. In the radiological follow-up in 40 patients (89%) findings were similar compared to the postoperative control. In five patients (11%) a loss of correction at a mean of 8 degrees degrees (Cobb angle) secondary to cage subsidence occurred. No breakage of the device or displacement of the instrumentation was seen. Overall the Frankel scale improved 0.65 points (p < 0.05) and the ODI improved 40.69 points (p < 0.05). CONCLUSIONS: In lumbar spine fractures of metastatic origin with stenosis, instability and/or neurological deficit, a single stage ventral decompression and instrumentation in patients with BMD >= 1.20 g/cm(2) should be considered. PMID- 21643686 TI - Update: schizophrenia across cultures. AB - The incidence of schizophrenia, as well as the symptoms, course, and outcomes for people so diagnosed seem to vary across some cultural contexts. The mechanisms by which cultural variations may protect one from or increase one's risk of developing schizophrenia remain unclear. Recent findings from transdisciplinary cross-cultural research, indicate ways that we may better understand how socioenvironmental and cultural variables interact with physiologic pathways relating psychosocial stress and psychotic symptoms, epigenetic changes, and people's use of culturally available tools to mitigate stress, in ways that may inform relevant, effective interventions for people diagnosed with psychotic disorders worldwide. PMID- 21643687 TI - Brown fat distribution in the chest wall of infants-normal appearance, distribution and evolution on CT scans of the chest. AB - BACKGROUND: While reviewing chest CT scans of infants, we repeatedly observed hyperdense enhancing tissue in the chest wall that is not well described in radiology literature. OBJECTIVE: This study was undertaken to describe the imaging features of this tissue in chest walls of infants. MATERIALS AND METHODS: CT scans of the chest conducted on all infants between April 2008 and October 2009 were retrospectively reviewed. CT studies with any deviation from normal radiation or contrast dose or those with chest wall anatomical distortion were excluded. RESULTS: One hundred eighty-eight infants were scanned, with 202 MDCTs, of which 180 (89.1%) received contrast agent. Fifty-four of 180 (30%) cases revealed focal areas of hyperdensity in various locations. All positive cases ranged between 2 days and 9 months of age. The areas of distribution of hyperdensity had excellent correlation with known areas of brown fat in the chest wall, known from nuclear medicine studies, and hence we concluded these to represent the same. CONCLUSION: Brown fat in the chest wall can be seen as enhancing tissue on contrast CT scans done on infants. This is a normal morphological component with the brown fat converting to normal fat. It is important to recognize it in the chest wall of infants to avoid misinterpretation. PMID- 21643688 TI - Label-free electrochemical aptasensor for thrombin detection based on the nafion@graphene as platform. AB - A sensitive label-free electrochemical aptasensor was successfully fabricated for thrombin detection with nafion@graphene as platform. With electrostatic interaction between nafion and methylene blue (MB), positive charged MB was successfully assembled on nafion@graphene modified electrode surface, which provided amounts of redox probes for electrochemical aptasensor. In the presence of thrombin, the thrombin aptamer (TBA) on the electrode surface would catch the target on the electrode interface, which made a barrier for electrons and inhibits the electro-transfer, resulting in the decreased differential pulse voltammetry signals of MB. As a result, the proposed approach showed a high sensitivity and a wider linearity to thrombin in the range 0.01-50 nM with a detection limit of 6 pM. PMID- 21643689 TI - Hypertrophic olivary degeneration after pontine hemorrhage. PMID- 21643690 TI - Role of Smads in TGFbeta signaling. AB - Transforming growth factor-beta (TGFbeta) is the prototype for a large family of pleiotropic factors that signal via heterotetrameric complexes of type I and type II serine/threonine kinase receptors. Important intracellular mediators of TGFbeta signaling are members of the Smad family. Smad2 and 3 are activated by C terminal receptor-mediated phosphorylation, whereafter they form complexes with Smad4 and are translocated to the nucleus where they, in cooperation with other transcription factors, co-activators and co-repressors, regulate the transcription of specific genes. Smads have key roles in exerting TGFbeta-induced programs leading to cell growth arrest and epithelial-mesenchymal transition. The activity and stability of Smad molecules are carefully regulated by a plethora of post-translational modifications, including phosphorylation, ubiquitination, sumoylation, acetylation and poly(ADP)-ribosylation. The Smad function has been shown to be perturbed in certain diseases such as cancer. PMID- 21643691 TI - Comparison of hormonal receptor and HER-2 status between breast primary tumours and relapsing tumours: clinical implications of progesterone receptor loss. AB - Differences in hormone receptor and HER-2 status between primary tumour and corresponding relapse could have a substantial impact on clinical management of patients. The aim of this study was to evaluate change in expression of hormone receptors and HER-2 status between primary tumour and corresponding local recurrence or distant metastasis. We analysed 140 primary tumours and related recurrent or metastatic samples. Hormone receptors status was evaluated by immunohistochemistry, while HER-2 status by immunohistochemistry and silver in situ hybridisation. A change in HER-2 was rare; 3.7% of cases by immunohistochemistry and only 0.7% by silver in situ hybridisation analysis. A change in estrogen and progesterone receptors was seen in 6.4% and 21.4% of cases, respectively. Estrogen receptor change was not affected by adjuvant therapy, whereas progesterone receptor was influenced by adjuvant chemotherapy associated to hormone therapy (P = 0.0005). A change in progesterone receptor was more frequent in distant metastases than in local recurrences (P = 0.03). In the setting of estrogen receptor positive tumours, patients with progesterone receptor loss in local recurrence had a statistically significant lower median metastasis free survival compared to others patients; progesterone receptor positive, 112 months; progesterone receptor negative, 24 months (P = 0.005). A change between primary tumour and corresponding relapse is frequent for progesterone receptor, infrequent for estrogen receptor and rare for HER-2. In cases with changes in HER-2, it is worthwhile reassessing HER-2 status with both immunohistochemistry and in situ hybridisation analysis. Progesterone receptor loss seems to be influenced by therapy and to correlate with a worse prognosis. PMID- 21643692 TI - Alpha-methylacyl-CoA racemase (AMACR) expression in epithelial ovarian cancer. AB - Alpha-methylacyl-CoA racemase (AMACR) is involved in the cellular metabolism of fatty acids. It is a prognostic factor in prostate and colorectal cancer. So far, little is known about its expression and prognostic role in ovarian cancer. We investigated the expression of AMACR in a total of 420 ovarian tumors (388 carcinomas, 32 borderline tumors) by immunohistochemistry on tissue microarrays of two independent patient cohorts. In both cohorts, cytoplasmic AMACR expression was identified in 11.8% (16/136) and 5.4% (13/239), respectively, of the ovarian carcinomas. In contrast, borderline tumors did not show any AMACR expression. AMACR expression was significantly associated with histological subtype, FIGO stage, and grade in one cohort and low estrogen receptor levels in the other cohort. In univariate analysis, AMACR expression was significantly associated with poor overall survival (log rank, p = 0.006) and an independent prognostic factor in a multivariate analysis (HR 3.3; CI 1.3-7.9; p = 0.008) but could not be verified in the second cohort. Unlike in other tumor entities, AMACR expression does not seem to have an unequivocal prognostic impact in ovarian cancer. The prevalence may limit the value of AMACR for the differential diagnosis between metastatic colorectal carcinomas and primary ovarian carcinomas, whereas the association with estrogen receptor expression deserves further studies. PMID- 21643693 TI - Novel missense mutation in the TMPRSS6 gene in a Japanese female with iron refractory iron deficiency anemia. AB - Iron-refractory iron deficiency anemia (IRIDA) is a rare autosomal-recessive disorder hallmarked by hypochromic microcytic anemia, low transferrin saturation, and unresponsiveness to oral iron with partial recovery after parenteral iron administration. The disease is caused by mutations in TMPRSS6 (transmembrane protease serine 6) that prevent inactivation of membrane-bound hemojuvelin, an activator of hepcidin transcription. To date, 38 cases have been characterized and reported in European countries and the United States. In this paper, we describe the first case of a Japanese female with IRIDA, who carried a novel mutation (K253E) in the CUB (complement factor C1r/C1s, urchin embryonic growth factor and bone morphogenetic protein 1) domain of the TMPRSS6 gene. PMID- 21643694 TI - Successful treatment of Hodgkin lymphoma in second trimester of pregnancy: feasibility of ABVD regimen. AB - A 34-year-old woman at 18 weeks of gestation during her third pregnancy was diagnosed with Hodgkin lymphoma stage IVA. She was treated with standard ABVD and showed good response. After three-and-a-half courses of ABVD, she spontaneously delivered a healthy baby girl at 41 weeks of gestation. She continued chemotherapy after delivery. A total of six courses of ABVD were given. Complete remission was obtained and has continued for 6 months post chemotherapy. Neither mother nor child developed any infections during the observation period. The child has also shown no physiological or developmental abnormalities. Here, we describe the feasibility and safety of the ABVD regimen for the treatment of Hodgkin lymphoma, and discuss the optimal treatment strategy. PMID- 21643695 TI - Auditory hallucinations and migraine of possible brainstem origin. AB - BACKGROUND: Concurrence of migraine and hallucinations is extremely rare and the underlying mechanism is poorly understood. METHODS: We report a 22-year-old man with migraine associated with auditory hallucinations. Concurrent psychotic illness has been excluded. RESULTS: Brain MR scans showed a stable, patchy FLAIR hyperintensity over the posterolateral aspect of the left cerebral peduncle, just below the level of the red nucleus. This was felt to represent an area of gliosis based on the interval stability over 19 months. There was absence of features for aggressive neoplasms, such as lesional high cellular turnover (choline/NAA ratio >1.0) or high cerebral blood volume on advanced MR imaging with MR spectroscopy and dynamic perfusion MR. EEG and brainstem auditory evoked potentials were unremarkable. CONCLUSIONS: To our knowledge, there are no reports to date on similar auditory hallucinations in adult migraine patients, as well as with associated MRI brainstem lesions. The peduncular lesion could represent a previous migrainous infarct, and a possible analogy can be drawn from the descriptions of peduncular hallucinosis. Brainstem lesions, particularly in the midbrain and pons, have rarely been associated with this condition. It has been postulated that the damage to ascending reticular systems or thalamocortical circuitry may contribute to its pathogenesis. PMID- 21643696 TI - Monitoring enzyme expression of a branched respiratory chain of corynebacterium glutamicum using an EGFP reporter gene. AB - To investigate the expressional control of branched respiratory chain complexes of the amino-acid producing bacterium Corynebacterium glutamicum according to growth conditions, the expression indexes of the ndh, sdh, qcrCAB, ctaCF, ctaD, ctaE, and cydAB genes were estimated under aerobic and microaerobic, and carbon rich and -poor conditions. The promoter region of each target gene was cloned upstream of the EGFP gene on expression vector pVK6, and the nine reporter constructs were transformed into C. glutamicum ssp. lactofermentum. The cytochrome content of cellular membranes obtained from each growth phase closely corresponded to the expression indexes based on EGFP fluorescence and cell density, indicating that this rapid and convenient method is suitable for analyzing the expression levels of respiratory chain complexes. Using this method, we demonstrated that a reciprocal change in the expression levels of cytochrome bd-type and aa (3)-type oxidases occurs when C. glutamicum cells are held in stationary phase for extended periods. PMID- 21643697 TI - Online monitoring of metabolism and morphology of peptide-treated neuroblastoma cancer cells and keratinocytes. AB - Antimicrobial peptides are promising anti-cancer agents with a unique mode of action. We established the usage of a chip-based sensor to monitor the dynamic interplay between cells on the chip and peptides and compared it with endpoint tests. Human neuroblastoma cancer cells and spontaneously immortalized non-cancer keratinocytes were perfused with representative peptides (NK-2, NK11, and melittin). The sensor system enabled continuous recording of cell layer impedance (adhesion/confluence), oxygen consumption (respiration) and extracellular acidification (glycolysis) and provided insights in cell damage, stress response and recovery. Cells responded differentially to peptide treatment. During perfusion, peptides accumulated on the cell surface until they reached a critical concentration. Preceding to cell death, melittin triggered glycolysis, suggesting stress response. NK-2 induced no change in energy metabolism, but led to an increase in impedance, i.e. a temporarily altered morphology, which appeared to be an excellent parameter to detect subtle structural changes of cell layers. PMID- 21643698 TI - How to juggle priorities? An interactive tool to provide quantitative support for strategic patient-mix decisions: an ophthalmology case. AB - An interactive tool was developed for the ophthalmology department of the Academic Medical Center to quantitatively support management with strategic patient-mix decisions. The tool enables management to alter the number of patients in various patient groups and to see the consequences in terms of key performance indicators. In our case study, we focused on the bottleneck: the operating room. First, we performed a literature review to identify all factors that influence an operating room's utilization rate. Next, we decided which factors were relevant to our study. For these relevant factors, two quantitative methods were applied to quantify the impact of an individual factor: regression analysis and computer simulation. Finally, the average duration of an operation, the number of cancellations due to overrun of previous surgeries, and the waiting time target for elective patients all turned out to have significant impact. Accordingly, for the case study, the interactive tool was shown to offer management quantitative decision support to act proactively to expected alterations in patient-mix. Hence, management can anticipate the future situation, and either alter the expected patient-mix or expand capacity to ensure that the key performance indicators will be met in the future. PMID- 21643700 TI - Soil cyanobacterial and microalgal diversity in dry mountains of Ladakh, NW Himalaya, as related to site, altitude, and vegetation. AB - Although phototrophic microbial communities are important components of soils in arid and semi-arid ecosystems around the world, the knowledge of their taxonomic composition and dependency on soil chemistry and vegetation is still fragmentary. We studied the abundance and the diversity of cyanobacteria and eukaryotic microalgae along altitudinal gradients (3,700-5,970 m) at four sites in the dry mountains of Ladakh (Little Tibet, Zanskar Mountains, and Eastern Karakoram), using epifluorescence. The effects of environmental factors (altitude, mountain range, and vegetation type) on soil physico-chemical parameters (pH; texture; organic matter, nitrogen, ammonia, and phosphorus contents; and concentration of chlorophylls and carotenoids) and on the composition and biovolume of phototrophs were tested by multivariate redundancy analysis and variance partitioning. Phototrophs were identified in all collected samples, and phototroph biovolume ranged from 0.08 to 0.32 mm(3) g(-1) dry weight. The dominant component was cyanobacteria, which represented 70.9% to 98.6% of the biovolume. Cyanobacterial species richness was low in that only 28 morphotypes were detected. The biovolume of Oscillatoriales consisted mainly of Phormidium spp. and Microcoleus vaginatus. The environmental factors accounted for 43.8% of the total variability in microbial and soil data, 20.6% of which was explained solely by mountain range, 7.0% by altitude, and 8.4% by vegetation type. Oscillatoriales prevailed in alpine meadows (which had relatively high organic matter and fine soil texture), while Nostocales dominated in the subnival zone and screes. Eukaryotic microalgae together with cyanobacteria in the order Chroococcales were mostly present in the subnival zone. We conclude that the high elevation, semiarid, and arid soils in Ladakh are suitable habitats for microbial phototrophic communities and that the differences in these communities are associated with site, altitude, and vegetation type. PMID- 21643699 TI - The Salmonella enterica pan-genome. AB - Salmonella enterica is divided into four subspecies containing a large number of different serovars, several of which are important zoonotic pathogens and some show a high degree of host specificity or host preference. We compare 45 sequenced S. enterica genomes that are publicly available (22 complete and 23 draft genome sequences). Of these, 35 were found to be of sufficiently good quality to allow a detailed analysis, along with two Escherichia coli strains (K 12 substr. DH10B and the avian pathogenic E. coli (APEC O1) strain). All genomes were subjected to standardized gene finding, and the core and pan-genome of Salmonella were estimated to be around 2,800 and 10,000 gene families, respectively. The constructed pan-genomic dendrograms suggest that gene content is often, but not uniformly correlated to serotype. Any given Salmonella strain has a large stable core, whilst there is an abundance of accessory genes, including the Salmonella pathogenicity islands (SPIs), transposable elements, phages, and plasmid DNA. We visualize conservation in the genomes in relation to chromosomal location and DNA structural features and find that variation in gene content is localized in a selection of variable genomic regions or islands. These include the SPIs but also encompass phage insertion sites and transposable elements. The islands were typically well conserved in several, but not all, isolates--a difference which may have implications in, e.g., host specificity. PMID- 21643701 TI - Co-fermentation of cellulose/xylan using engineered industrial yeast strain OC-2 displaying both beta-glucosidase and beta-xylosidase. AB - We constructed a recombinant industrial Saccharomyces cerevisiae yeast strain OC2 AXYL2-ABGL2-Xyl2 by inserting two copies of the beta-glucosidase (BGL) and beta xylosidase (XYL) genes, and a gene cassette for xylose assimilation in the genome of yeast strain OC-2HUT. Both BGL and XYL were expressed on the yeast cell surface with high enzyme activities. Using OC2-AXYL2-ABGL2-Xyl2, we performed ethanol fermentation from a mixture of powdered cellulose (KC-flock) and Birchwood xylan, with the additional supplementation of a 30-g/l Trichoderma reesei cellulase complex mixture. The ethanol yield (gram per gram of added cellulases) of the strain OC2-AXYL2-ABGL2-Xyl2 increased approximately 2.5-fold compared to that of strain OC2-Xyl2, which lacked beta-glucosidase and beta xylosidase activities. Notably, the concentration of additional T. reesei cellulase was reduced from 30 to 24 g/l without affecting ethanol production. The BGL- and XYL-displaying industrial yeast of the strain OC2-AXYL2-ABGL2-Xyl2 represents a promising yeast for reducing cellulase consumption of ethanol fermentation from lignocellulosic biomass by compensating for the inherent weak BGL and XYL activities of T. reesei cellulase complexes. PMID- 21643702 TI - Homo-D-lactic acid production from mixed sugars using xylose-assimilating operon integrated Lactobacillus plantarum. AB - In order to achieve efficient D-lactic acid fermentation from a mixture of xylose and glucose, the xylose-assimilating xylAB operon from Lactobacillus pentosus (PXylAB) was introduced into an L-lactate dehydrogenase gene (ldhL1)-deficient Lactobacillus plantarum (DeltaldhL1-xpk1::tkt-Deltaxpk2) strain in which the phosphoketolase 1 gene (xpk1) was replaced with the transketolase gene (tkt) from Lactococcus lactis, and the phosphoketolase 2 (xpk2) gene was deleted. Two copies of xylAB introduced into the genome significantly improved the xylose fermentation ability, raising it to the same level as that of DeltaldhL1 xpk1::tkt-Deltaxpk2 harboring a xylAB operon-expressing plasmid. Using the two copy xylAB integrated strain, successful homo-D-lactic acid production was achieved from a mixture of 25 g/l xylose and 75 g/l glucose without carbon catabolite repression. After 36-h cultivation, 74.2 g/l of lactic acid was produced with a high yield (0.78 g per gram of consumed sugar) and an optical purity of D-lactic acid of 99.5%. Finally, we successfully demonstrated homo-D lactic acid fermentation from a mixture of three kinds of sugar: glucose, xylose, and arabinose. This is the first report that describes homo-D-lactic acid fermentation from mixed sugars without carbon catabolite repression using the xylose-assimilating pathway integrated into lactic acid bacteria. PMID- 21643703 TI - Substrate promiscuity of secondary metabolite enzymes: prenylation of hydroxynaphthalenes by fungal indole prenyltransferases. AB - Fungal prenyltransferases of the dimethylallyltryptophan synthase (DMATS) superfamily share no sequence, but structure similarity with the prenyltransferases of the CloQ/NphB group. The members of the DMATS superfamily have been reported to catalyze different prenylations of diverse indole or tyrosine derivatives, while some members of the CloQ/NphB group used hydroxynaphthalenes as prenylation substrates. In this study, we report for the first time the prenylation of hydroxynaphthalenes by the members of the DMATS superfamily. Three tryptophan-containing cyclic dipeptide prenyltransferases (AnaPT, CdpNPT and CdpC3PT), one tryptophan C7-prenyltransferase and one tyrosine O-prenyltransferase (SirD) were incubated with naphthalene and 11 derivatives. The enzyme activity and preference of the tested prenyltransferases towards hydroxynaphthalenes differed clearly from each other. For an accepted substrate, however, different enzymes produced usually the same major prenylation product, i.e. with a regular C-prenyl moiety at para- or ortho-position to a hydroxyl group. Regularly, O-prenylated and diprenylated derivatives were also identified as enzyme products of substrates with low conversion rates and regioselectivity. This was unequivocally proven by mass spectrometry and nuclear magnetic resonance analyses. The K (M) values and turnover numbers (k (cat)) of the enzymes towards selected hydroxynaphthalenes were determined to be in the range of 0.064-2.8 mM and 0.038-1.30 s(-1), respectively. These data are comparable to those obtained using indole derivatives. The results presented in this study expanded the potential usage of the members of the DMATS superfamily for production of prenylated derivatives including hydroxynaphthalenes. PMID- 21643704 TI - Bacterial production of free fatty acids from freshwater macroalgal cellulose. AB - The predominant strategy for using algae to produce biofuels relies on the overproduction of lipids in microalgae with subsequent conversion to biodiesel (methyl-esters) or green diesel (alkanes). Conditions that both optimize algal growth and lipid accumulation rarely overlap, and differences in growth rates can lead to wild species outcompeting the desired lipid-rich strains. Here, we demonstrate an alternative strategy in which cellulose contained in the cell walls of multicellular algae is used as a feedstock for cultivating biofuel producing microorganisms. Cellulose was extracted from an environmental sample of Cladophora glomerata-dominated periphyton that was collected from Lake Mendota, WI, USA. The resulting cellulose cake was hydrolyzed by commercial enzymes to release fermentable glucose. The hydrolysis mixture was used to formulate an undefined medium that was able to support the growth, without supplementation, of a free fatty acid (FFA)-overproducing strain of Escherichia coli (Lennen et. al 2010). To maximize free fatty acid production from glucose, an isopropyl beta-D-1 thiogalactopyranoside (IPTG)-inducible vector was constructed to express the Umbellularia californica acyl-acyl carrier protein (ACP) thioesterase. Thioesterase expression was optimized by inducing cultures with 50 MUM IPTG. Cell density and FFA titers from cultures grown on algae-based media reached 50% of those (~90 MUg/mL FFA) cultures grown on rich Luria-Bertani broth supplemented with 0.2% glucose. In comparison, cultures grown in two media based on AFEX pretreated corn stover generated tenfold less FFA than cultures grown in algae based media. This study demonstrates that macroalgal cellulose is a potential carbon source for the production of biofuels or other microbially synthesized compounds. PMID- 21643705 TI - Improved phloroglucinol production by metabolically engineered Escherichia coli. AB - Phloroglucinol is a valuable chemical which has been successfully produced by metabolically engineered Escherichia coli. However, the low productivity remains a bottleneck for large-scale application and cost-effective production. In the present work, we cloned the key biosynthetic gene, phlD (a type III polyketide synthase), into a bacterial expression vector to produce phloroglucinol in E. coli and developed different strategies to re-engineer the recombinant strain for robust synthesis of phloroglucinol. Overexpression of E. coli marA (multiple antibiotic resistance) gene enhanced phloroglucinol resistance and elevated phloroglucinol production to 0.27 g/g dry cell weight. Augmentation of the intracellular malonyl coenzyme A (malonyl-CoA) level through coordinated expression of four acetyl-CoA carboxylase (ACCase) subunits increased phloroglucinol production to around 0.27 g/g dry cell weight. Furthermore, the coexpression of ACCase and marA caused another marked improvement in phloroglucinol production 0.45 g/g dry cell weight, that is, 3.3-fold to the original strain. Under fed-batch conditions, this finally engineered strain accumulated phloroglucinol up to 3.8 g/L in the culture 12 h after induction, corresponding to a volumetric productivity of 0.32 g/L/h. This result was the highest phloroglucinol production to date and showed promising to make the bioprocess economically feasible. PMID- 21643706 TI - Correlation between pellet morphology and glycopeptide antibiotic balhimycin production by Amycolatopsis balhimycina DSM 5908. AB - Actinomycetes, a class of filamentous bacteria, are an important source of several industrially relevant secondary metabolites. Several environmental factors including the media composition affect both biomass growth and product formation. Likewise, several studies have shown that environmental factors cause changes in cellular morphology. However, the relationship between morphology and product formation is not well understood. In this study, we first characterized the effect of varying concentrations of phosphate and ammonia in defined media on pellet morphology for an actinomycete Amycolatopsis balhimycina DSM 5908, which produces balhimycin, a glycopeptide antibiotic. Our results show that higher balhimycin productivity is correlated with the following morphological features: (1) higher pellet fraction in the biomass, (2) small elongated pellets, and (3) shorter filaments in hyphal growth in the periphery of the pellets. The correlation between morphology and product formation was also observed in industrially relevant complex media. Although balhimycin production starts after 72 h with maximum production around 168 h, the morphological changes in pellets are observed as early as 24 h after commencing of the batch. Therefore, morphology may be used as an early predictor of the end-of-batch productivity. We argue that a similar strategy can be developed for other strains where morphological indicators may be used as a batch monitoring tool. PMID- 21643707 TI - Production of bioactive sheep beta-defensin-1 in Pichia pastoris. AB - Previous research has shown that sheep beta-defensin-1 (sBD-1), a small cationic peptide with a broad range of antimicrobial activities, could inhibit the growth of both Gram-positive and Gram-negative bacteria as well as that of fungi. In order to increase the yield of current ovine defensin purification methods, mature sBD-1 (msBD-1) was added with a 6-His tag on the C-terminus (msBD-1-T) and expressed in Pichia pastoris in the presented work. The msBD-1 and msBD-1-T were expressed in the Pichia pastoris. Both msBD-1 and msBD-1-T were purification, and the two peptides were used to inhibit Escherichia coli, Staphylococcus aureus, Proteus vulgaris, Pseudomonas aeruginosa, and Shigella flexneri. The antimicrobial activity of the 6-His tagged msBD-1-T peptide was not significantly different from that of the native msBD-1 peptide. The two peptides could inhibit the growth of Escherichia coli, Staphylococcus aureus, Proteus vulgaris, Pseudomonas aeruginosa, and Shigella flexneri with equal efficiency as well as chemoattractant function. In addition, the yield of purified 6-His-tagged msBD-1 was greater than that of msBD-1. The presented method might be a more efficient approach to produce bioactive sBD-1. PMID- 21643708 TI - Enzymatic deinking of secondary fibers: cellulases/hemicellulases versus laccase mediator system. AB - The use of enzymes has been suggested as an environmentally friendly alternative to complement conventional chemical deinking in the recycling of recovered paper. This study compares the use of cellulases/hemicellulases versus the laccase mediator system for deinking printed fibers from newspapers and magazines. For this purpose, two commercial enzyme preparations with endoglucanase and endoxylanase activities (Viscozyme Wheat from Aspergillus oryzae and Ultraflo L from Humicola insolens, Novozymes) and a commercial laccase (NS51002 from Trametes villosa, Novozymes), the latter in the presence of synthetic or natural (lignin-related) mediators, were evaluated. The enzymatic treatments were studied at the laboratory scale using a standard chemical deinking sequence consisting of a pulping stage; an alkaline stage using NaOH, sodium silicate and fatty acid soap; and a bleaching stage using hydrogen peroxide. The handsheets were then prepared and their brightness, residual ink concentration, and strength properties were measured. Among the different enzymatic treatments assayed, both carbohydrate hydrolases were found to deink the secondary fibers more efficiently. Brightness increased up to 3-4% ISO on newspaper fibers, being Ultraflo 20% more efficient in the ink removal. Up to 2.5% ISO brightness increase was obtained when magazine fibers were used, being Viscozyme 9% more efficient in the ink removal. Regarding the laccase-mediator system, alone or in combination with carbohydrate hydrolases, it was ineffective in deinking both newspaper and magazine fibers, resulting in pulps with worse brightness and residual ink concentration values. However, pulp deinking by the laccase-mediator system was displayed when secondary fibers from printed cardboard were used, obtaining up to 3% ISO brightness increase and lower residual ink concentrations. PMID- 21643709 TI - Identification of a xylose reductase gene in the xylose metabolic pathway of Kluyveromyces marxianus NBRC1777. AB - Kluyveromyces marxianus is thermotolerant yeast that is able to utilize a wider range of substrates and has greater thermal tolerance than most other yeast species. K. marxianus can assimilate xylose, but its ability to produce ethanol from xylose in oxygen-limited environments is poor. In the present study, the K. marxianus xylose reductase (KmXR) gene (Kmxyl1) was cloned and the recombinant enzyme was characterized to clarify the factors that limit xylose fermentation in K. marxianus NBRC1777. KmXR is a key enzyme in the xylose metabolism of K. marxianus, which was verified by disruption of the Kmxyl1 gene. The Km of the recombinant KmXR for NADPH is 65.67 MUM and KmXR activity is 1.295 U/mg, which is lower than those of most reported yeast XRs, and the enzyme has no activity with coenzyme NADH. This result demonstrates that the XR from K. marxianus is highly coenzyme specific; combined with the extremely low XDH activity of K. marxianus with NADP+, the limitation of xylose fermentation is due to a redox imbalance under anaerobic conditions and low KmXR activity. PMID- 21643710 TI - Electrochemical behavior of the 316L steel type in a marine culture of microalgae (Porphyridium purpureum) under the 12/12 h photoperiod and effect of different working electrode exposure conditions on the biofilm-metal interface. AB - The industrial crops of microalgae use processes calling upon the presence of parts of metal nature such as steel 316L type. The goal of this study is to test the electrochemical behavior of this material in a marine culture of microalgae. Porphyridium purpureum was used under a photoperiod of alternation darkness/light 12/12 h, in order to apprehend the problems of biocorrosion involved in the biofouling. The evolution of the free potential of corrosion, according to the position of the samples and for different surface roughness, observations of the surface quality under the electron microscope with sweeping were carried out. The results showed that, overall, the strain P. purpureum does not have a corrosive effect on the 316L. The free potential of corrosion lies between -0.307 and 0.005 V(SCE). The adhesion of the cells seems stronger on the interface air/solid of the half-plunged sample with surface grit polished 1,000, confirmed by the presence of biofilm on the air part. The photoperiod acts on the evolution of the generated free potential of corrosion of the one 24-h period oscillation. Furthermore, the samples plunged horizontally lead to a stabilizing effect on the potential of free corrosion. PMID- 21643711 TI - Working memory load and the vigilance decrement. AB - In this study, we examined the impact of concurrent verbal and spatial working memory demands on performance on an alpha-numeric successive target detection task. Seven hundred and forty-five participants performed a target detection task while simultaneously performing either a spatial or a verbal working memory task or they performed matched no-memory control tasks. The vigilance decrement, both an increase in target detection response times and a decrease in perceptual sensitivity A' to target stimuli over time, was exacerbated by concurrent working memory load. The spatial and verbal working memory loads both impacted vigilance performance suggesting utilization of common executive resources. Overall, these results support the view that the vigilance decrement results from high cognitive resource demands (e.g., hard work), not from cognitive under-load (e.g., boredom or mindlessness). PMID- 21643712 TI - Pointing with the wrist: a postural model for Donders' law. AB - The central nervous system uses stereotypical combinations of the three wrist/forearm joint angles to point in a given (2D) direction in space. In this paper, we first confirm and analyze this Donders' law for the wrist as well as the distributions of the joint angles. We find that the quadratic surfaces fitting the experimental wrist configurations during pointing tasks are characterized by a subject-specific Koenderink shape index and by a bias due to the prono-supination angle distribution. We then introduce a simple postural model using only four parameters to explain these characteristics in a pointing task. The model specifies the redundancy of the pointing task by determining the one-dimensional task-equivalent manifold (TEM), parameterized via wrist torsion. For every pointing direction, the torsion is obtained by the concurrent minimization of an extrinsic cost, which guarantees minimal angle rotations (similar to Listing's law for eye movements) and of an intrinsic cost, which penalizes wrist configurations away from comfortable postures. This allows simulating the sequence of wrist orientations to point at eight peripheral targets, from a central one, passing through intermediate points. The simulation first shows that in contrast to eye movements, which can be predicted by only considering the extrinsic cost (i.e., Listing's law), both costs are necessary to account for the wrist/forearm experimental data. Second, fitting the synthetic Donders' law from the simulated task with a quadratic surface yields similar fitting errors compared to experimental data. PMID- 21643713 TI - Proprioceptive integration and body representation: insights into dancers' expertise. AB - The experience of the body as a single coherent whole is based on multiple local sensory signals, integrated across different sensory modalities. We investigated how local information is integrated to form a single body representation and also compared the contribution of proprioceptive and visual information both in expert dancers and non-dancer controls. A number of previous studies have focused on individual differences in proprioceptive acuity at single joints and reported inconsistent findings. We used the established endpoint position matching task to measure absolute and directional errors in matching the position of one hand with the other. The matching performance was tested in three different conditions, which involved different information about the target position: only proprioceptive information from a 'target' hand which could be either the left or the right, only visual information, or both proprioceptive and visual information. Differences in matching errors between these sensory conditions suggested that dancers show better integration of local proprioceptive signals than non-dancers. The dancers also relied more on proprioception when both proprioceptive and visual information about hand position were present. PMID- 21643714 TI - Looming sounds enhance orientation sensitivity for visual stimuli on the same side as such sounds. AB - Several recent multisensory studies show that sounds can influence visual processing. Some visual judgments can be enhanced for visual stimuli near a sound occurring around the same time. A recent TMS study (Romei et al. 2009) indicates looming sounds might influence visual cortex particularly strongly. But unlike most previous behavioral studies of possible audio-visual exogenous effects, TMS phosphene thresholds rather than judgments of external visual stimuli were measured. Moreover, the visual hemifield assessed relative to the hemifield of the sound was not varied. Here, we compared the impact of looming sounds to receding or "static" sounds, using auditory stimuli adapted from Romei et al. (2009), but now assessing any influence on visual orientation discrimination for Gabor patches (well-known to involve early visual cortex) when appearing in the same hemifield as the sound or on the opposite side. The looming sounds that were effective in Romei et al. (2009) enhanced visual orientation sensitivity (d') here on the side of the sound, but not for the opposite hemifield. This crossmodal, spatially specific effect was stronger for looming than receding or static sounds. Similarly to Romei et al. (2009), the differential effect for looming sounds was eliminated when using white noise rather than structured sounds. Our new results show that looming structured sounds can specifically benefit visual orientation sensitivity in the hemifield of the sound, even when the sound provides no information about visual orientation itself. PMID- 21643715 TI - The effect of head roll on perceived auditory zenith. AB - We studied the influence of static head roll on the perceived auditory zenith in head-centred and world-centred coordinates. Subjects sat either upright, or with their head left/right rolled sideways by about 35 degrees relative to gravity, whilst judging whether a broadband sound was heard left or right from the head centred or world-centred zenith. When upright, these reference frames coincide. Results show that subjects judged the zenith accurately within different planes, although response variability increased for the midsagittal plane. With the head rolled, head-centred auditory zenith shifted by the same amount and was located as accurately as for upright, indicating unaltered localisation cues by head-on body roll. Interestingly, when judging world-centred zenith subjects made large systematic errors (10-15 degrees ) in the direction of head roll, and response variability increased, which resembles the visual Aubert effect. These results demonstrate a significant influence of the vestibular-collic system on auditory spatial awareness, which sheds new light on the mechanisms underlying multisensory integration and spatial updating in sound localisation behaviour. PMID- 21643716 TI - An ERP study on whether the P600 can reflect the presence of unexpected phonology. AB - In this study, we conducted a new approach to determine whether the P600 could reflect the presence of unexpected phonology. In the experiment, the critical Chinese characters in the poems are either original or substituted by homophones or synonyms. The N400 effect is observed only under the homophonic condition. Late positive shifts are revealed under both homophonic and synonymous conditions, but the amplitude under the homophonic condition is smaller than that under the synonymous condition. The observation of the N400 effect under the homophonic condition indicates that semantic processing occurs in Chinese poem comprehension, while the P600 effect elicited under the homophonic condition is a reflection of unfulfilled semantic expectations. The existence of the late positivity under the synonymous condition reveals that P600 can reflect the presence of unexpected phonology. Interestingly, late positivities under the two conditions both consist of two parts. A discussion ensues wherein these two parts of the late positive shift relate, respectively, to monitoring and resolution processes in language comprehension. PMID- 21643717 TI - Postural control in response to a perturbation: role of vision and additional support. AB - The purpose of the study was to investigate the availability of vision and additional support on anticipatory (APA) and compensatory (CPA) postural adjustments and their interaction. Eight healthy adults were exposed to external perturbations induced at the shoulder level while standing with and without holding onto a walker in full vision and while blindfolded. Electrical activity of the trunk and leg muscles and center of pressure (COPAP) displacement were recorded and quantified within the time intervals typical of APA and CPA. The results showed that with full vision, there was no difference in both APA and CPA in standing with and without holding onto a walker. With subjects holding onto a walker, CPA in standing blindfolded were comparable to CPA in full vision; this was seen in changes in the electrical activity of most of the muscles at the individual muscle, joint, and the muscle group levels as well as in COPAP displacement. The findings suggest that (1) in conditions where vision is available, vision overrules simultaneously available proprioceptive information from the support, (2) while in conditions where vision is not available, proprioceptive information from the support or support itself could be substituted for vision. It is possible to suggest that using a non-stabilizing support could be a valuable strategy to improve postural control when visual information is not available or compromised. PMID- 21643718 TI - Deficits in inhibitory control and conflict resolution on cognitive and motor tasks in Parkinson's disease. AB - Recent imaging studies in healthy controls with a conditional stop signal reaction time (RT) task have implicated the subthalamic nucleus (STN) in response inhibition and the pre-supplementary motor area (pre-SMA) in conflict resolution. Parkinson's disease (PD) is characterized by striatal dopamine deficiency and overactivity of the STN and underactivation of the pre-SMA during movement. We used the conditional stop signal RT task to investigate whether PD produced similar or dissociable effects on response initiation, response inhibition and response initiation under conflict. In addition, we also examined inhibition of prepotent responses on three cognitive tasks: the Stroop, random number generation and Hayling sentence completion. PD patients were impaired on the conditional stop signal reaction time task, with response initiation both in situations with or without conflict and response inhibition all being significantly delayed, and had significantly greater difficulty in suppressing prepotent or habitual responses on the Stroop, Hayling and random number generation tasks relative to controls. These results demonstrate the existence of a generalized inhibitory deficit in PD, which suggest that PD is a disorder of inhibition as well as activation and that in situations of conflict, executive control over responses is compromised. PMID- 21643719 TI - Attention does more than modulate suppressive interactions: attending to multiple items. AB - Directing attention to a visual item enhances its representations, making it more likely to guide behavior (Corbetta et al. 1991). Attention is thought to produce this enhancement by biasing suppressive interactions among multiple items in visual cortex in favor of the attended item (e.g., Desimone and Duncan 1995; Reynolds and Heeger 2009). We ask whether target enhancement and modulation of suppressive interactions are in fact inextricably linked or whether they can be decoupled. In particular, we ask whether simultaneously directing attention to multiple items may be one means of dissociating the influence of attention related enhancement from the effects of inter-item suppression. When multiple items are attended, suppressive interactions in visual cortex limit the effectiveness with which attention may act on their representations, presumably because "biasing" the interactions in favor of a single item is no longer possible (Scalf and Beck 2010). In this experiment, we directly investigate whether applying attention to multiple competing stimulus items has any influence on either their evoked signal or their suppressive interactions. Both BOLD signal evoked by the items in V4 and behavioral responses to those items were significantly compromised by simultaneous presentation relative to simultaneous presentation, indicating that when the items appeared at the same time, they interacted in a mutually suppressive manner that compromised their ability to guide behavior. Attention significantly enhanced signal in V4. The attentional status of the items, however, had no influence on the suppressive effects of simultaneous presentation. To our knowledge, these data are the first to explicitly decouple the effects of top-down attention from those of inter-item suppression. PMID- 21643720 TI - Inhibition of phosphodiesterases leads to prevention of the mitochondrial permeability transition pore opening and reperfusion injury in cardiac H9c2 cells. AB - PURPOSE: We tested if inhibition of phosphodiesterases (PDEs) with IBMX (1-methyl 3-isobutylxanthine) can modulate the mitochondrial permeability transition pore (mPTP) opening by inactivating glycogen synthase kinase 3beta (GSK-3beta). METHODS: H9c2 cells were exposed to 600 MUM H(2)O(2) for 20 min to cause the mPTP opening. Mitochondrial membrane potential (DeltaPsim) was assessed by imaging cells loaded with tetramethylrhodamine ethyl ester (TMRE). Cell viability was measured with propidium iodide (PI) fluorometry using a fluorescence reader. Ischemia/reperfusion injury was induced by exposing cells to ischemic solution for 90 min followed by 30 min of reperfusion. RESULTS: IBMX reduced loss of DeltaPsim caused by H(2)O(2), indicating that inhibition of PDEs can prevent the mPTP opening. However, IBMX could not inhibit the pore opening in cells transfected with the constitutively active GSK-3beta (GSK-3beta-S9A) mutant, suggesting a critical role of GSK-3beta in the action of IBMX. IBMX also reduced reperfusion injury in a GSK-3beta dependent manner. In support, IBMX increased GSK-3beta phosphorylation at Ser(9), an effect that was reversed by both the PKA inhibitor H89 and the PKG inhibitor KT5823. In support, IBMX activated both PKA and PKG. IBMX failed to prevent the loss of DeltaPsim in the presence of H89 or PKA siRNA. Similarly, both KT5823 and PKG siRNA reversed the protective effect of IBMX. CONCLUSION: Inhibition of PDEs prevents the mPTP opening by inactivating GSK-3beta through PKA and PKG. GSK-3beta is a common downstream target of PKA and PKG. Inhibition of PDEs may be a useful approach to prevent reperfusion injury. PMID- 21643721 TI - Is the peripheral retina an important site for myopic development? [Liu Y, Wildsoet C (2011) The effectof two-zone concentric bifocal spectacle lenses on refractive error development and eye growth in young chicks. Invest Ophthalmol Vis Sci 52(2):1078-1086]. PMID- 21643722 TI - Reduced retinal blood flow-velocity in severe hyperlipidemia measured by the retinal function imager. PMID- 21643723 TI - Incisor enamel formation is impaired in transgenic rats overexpressing the type III NaPi transporter Slc20a1. AB - Inorganic phosphate (Pi) is required in many biological processes, including signaling cascades, skeletal development, tooth mineralization, and nucleic acid synthesis. Recently, we showed that Pi transport in osteoblasts, mediated by Slc20a1, a member of the type III sodium-dependent phosphate transporter family, is indispensable for osteoid mineralization in rapidly growing rat bone. In addition, we found that bone mineral density decreased slightly with dysfunction of Pi homeostasis in aged transgenic rats overexpressing mouse Slc20a1 (Slc20a1 Tg). Bone and tooth share certain common molecular features, and thus, we focused on tooth development in Slc20a1-Tg mandibular incisors in order to determine the role of Slc20a1 in tooth mineralization. Around the time of weaning, there were no significant differences in serologic parameters between wild-type and Slc20a1 Tg rats. However, histological analysis showed that Slc20a1-Tg ameloblasts formed clusters in the papillary layer during the maturation stage as early as 4 weeks of age. These pathologies became more severe with age and included the formation of cyst-like or multilayer ameloblast structures, accompanied by a chalky white appearance with abnormal attrition and fracture. Hyperphosphatemia was also observed in aging Slc20a1-Tg rats. Micro-computed tomography and electron probe microanalysis revealed impairments in enamel, such as delayed mineralization and hypomineralization. Our results suggest that enamel formation is sensitive to imbalances in Pit1-mediated cellular function as seen in bone, although these processes are under the control of systemic Pi homeostasis. PMID- 21643724 TI - An atypical degenerative osteoarthropathy in Hyp mice is characterized by a loss in the mineralized zone of articular cartilage. AB - Patients with X-linked hypophosphatemia (XLH) develop enthesophytes and osteophytes secondary to articular cartilage degeneration and together are the primary cause of morbidity in adult patients so afflicted. We have previously characterized the enthesopathy in Hyp mice, a murine model of XLH. We now extend these studies to the synovial joint in order to characterize potential cellular changes in articular cartilage that may predispose patients to the osteoarthropathy of XLH. We report that, despite highly elevated levels of alkaline phosphatase activity throughout articular cartilage, there is a complete loss in the mineralized zone of articular cartilage as assessed by von Kossa staining of mineral and as quantified by EPIC-microCT analysis and evidence of vascular invasion. We also identify the downregulation of extracellular matrix (ECM) factors identified as regulators of terminally differentiated mineralizing articular chondrocytes. There is also a striking increase in the histochemical staining of sulfated proteoglycans, a change that may reflect the loss of a transitional tissue that reduces mechanical stress at the interface between cartilage and subchondral bone. The failure of mineralizing articular chondrocytes to develop in the hypophosphatemic state suggests that phosphate may be a key regulator of chondrocyte mineralization. Accordingly, we find that the appropriate zonal arrangement and phenotypic markers of articular cartilage are significantly reestablished by phosphate-replacement therapy. Given the turnover and maintenance of articular cartilage ECM, the identification of early and abnormal cellular changes unique to XLH will undoubtedly aid in a more effective management of this disease to minimize the onset of degenerative osteoarthropathy. PMID- 21643725 TI - Working conditions of Brazilian immigrants in Massachusetts. AB - Brazilian immigration to Massachusetts and other states in the US grew significantly in the last two decades. There is a lack of data about the working conditions and health and safety hazards faced by Brazilian immigrant workers. We surveyed over 500 workers in Eastern Massachusetts through a community-based participatory research project to explore occupational and immigration factors that may represent a risk to the health of Brazilian immigrant workers, who mostly work in the construction, housecleaning, and food services segments of the state labor force. Our pilot study suggests that Brazilian immigrant workers are exposed to chemical, ergonomic, physical, and psychosocial job hazards and have experienced a variety of health symptoms that may be associated with these work environment exposures. Since most Brazilian workers have not received proper training to recognize the hazards, there is an urgent need for the implementation of culturally adequate training programs and enforcement of safety and health regulations to prevent occupational injuries and fatalities. PMID- 21643727 TI - Adoption of safety eyewear among citrus harvesters in rural Florida. AB - The community-based prevention marketing program planning framework was used to adapt an evidence-based intervention to address eye injuries among Florida's migrant citrus harvesters. Participant-observer techniques, other direct observations, and individual and focus group interviews provided data that guided refinement of a safety eyewear intervention. Workers were attracted to the eyewear's ability to minimize irritation, offer protection from trauma, and enable work without declines in productivity or comfort. Access to safety glasses equipped with worker-designed features reduced the perceived barriers of using them; deployment of trained peer-leaders helped promote adoption. Workers' use of safety glasses increased from less than 2% to between 28% and 37% in less than two full harvesting seasons. The combination of formative research and program implementation data provided insights for tailoring an existing evidence-based program for this occupational community and increase potential for future dissemination and worker protection. PMID- 21643728 TI - HIV knowledge among Canadian-born and sub-Saharan African-born patients living with HIV. AB - Research has revealed differences on scales measuring HIV knowledge between individuals from various ethnic backgrounds and cultures. Few studies have examined this knowledge with immigrant populations and persons living with HIV. This study examined HIV knowledge among persons living with HIV who were either born in Canada or in sub-Saharan Africa and, for comparison, in a sample of college students. All participants were residing in Canada. Participants completed questionnaires measuring demographic variables, sexual health behaviour, and HIV status, treatment, and knowledge. Canadian-born patients living with HIV were more likely to be older and male than the other groups. On average, patients living with HIV were diagnosed 6.4 years ago, and 80% reported having current or previous experience taking HIV medications. After adjusting for age and gender, significant differences were found between the groups on the Brief HIV Knowledge Questionnaire. Canadian-born persons living with HIV (n = 110) scored higher than sub-Saharan African-born patients (n = 23) and college students (n = 81); mean percentage correct was 86, 70, and 62%, respectively (P < .01). These results suggested that ongoing HIV education is needed for all groups, and that additional tailored and targeted educational interventions are needed to address important gaps in knowledge among persons living with HIV patients originating from Africa and among college students. PMID- 21643729 TI - The expression patterns of Nogo-A, myelin associated glycoprotein and oligodendrocyte myelin glycoprotein in the retina after ocular hypertension: the expression of myelin proteins in the retina in glaucoma. AB - Nogo-A, a major myelin inhibitory protein, inhibits axon growth and synaptic function in the central nervous system. Glaucoma is a progressive neuropathy as a result of retinal ganglion cell (RGC) death. Synaptic degeneration is thought to be an early pathology of neurodegeneration in glaucoma and precedes RGC loss. Here experimental ocular hypertension model was induced in adult rats with laser coagulation of the episcleral and limbal veins. The expression of Nogo-A, myelin associated glycoprotein (MAG) and oligodendrocyte myelin glycoprotein (OMgp) in the retina was investigated using immunohistochemistry and Western blotting. We found that Nogo-A was expressed in the RGCs and upregulated after the induction of ocular hypertension. OMgp was only expressed in the inner plexiform layer. There was no MAG expression in the retina. Our data provided, for the first time, the expression patterns of three myelin proteins in the adult retina and suggested an important role of Nogo-A in the RGC death and synaptic degeneration in glaucoma. PMID- 21643730 TI - Stability analysis of vascular endothelial growth factor in cerebrospinal fluid. AB - Vascular endothelial growth factor (VEGF) is a promising biological marker and prognostic indicator in many neurological diseases. Although VEGF concentrations in plasma and cerebrospinal fluid (CSF) are increasingly reported, CSF-VEGF stability pre- and during-assay procedures is seldom evaluated. In the current study, we investigated VEGF variability and stability in CSF related to sample preparation, storage, and routine experimental procedures. Results showed that contaminant cell breakdown or aggregation can occur gradually before sample processing. However, after the removal of contaminant cell components, CSF-VEGF levels did not show significant changes in samples incubated at room temperature for 5 h, thawed/refrozen for 6 cycles. Samples preserved at -80 degrees C for up to 7 years continued to show measurable levels. Since some cellular components such as platelets contain a large amount of releasable VEGF, we conclude that CSF samples should be processed as soon as possible to carefully remove all cellular components and prevent possible consequent release of VEGF into CSF. After centrifugation to remove cellular contents, VEGF in CSF was relatively stable during routine experimental procedures and storage. PMID- 21643731 TI - Pseudomonas siderophores in the sputum of patients with cystic fibrosis. AB - The lungs of patients with cystic fibrosis become chronically infected with the bacterium Pseudomonas aeruginosa, which heralds progressive lung damage and a decline in health. Iron is a crucial micronutrient for bacteria and its acquisition is a key factor in infection. P. aeruginosa can acquire this element by secreting pyoverdine and pyochelin, iron-chelating compounds (siderophores) that scavenge iron and deliver it to the bacteria. Siderophore-mediated iron uptake is generally considered a key factor in the ability of P. aeruginosa to cause infection. We have investigated the amounts of pyoverdine in 148 sputum samples from 36 cystic fibrosis patients (30 infected with P. aeruginosa and 6 as negative controls). Pyoverdine was present in 93 samples in concentrations between 0.30 and 51 MUM (median 4.6 MUM) and there was a strong association between the amount of pyoverdine and the number of P. aeruginosa present. However, pyoverdine was not present, or below the limits of detection (~0.3 MUM), in 21 sputum samples that contained P. aeruginosa. Pyochelin was also absent, or below the limits of detection (~1 MUM), in samples from P. aeruginosa-infected patients with little or no detectable pyoverdine. Our data show that pyoverdine is an important iron-scavenging molecule for P. aeruginosa in many cystic fibrosis patients, but other P. aeruginosa iron-uptake systems must be active in some patients to satisfy the bacterial need for iron. PMID- 21643732 TI - The cognitive paradigm ontology: design and application. AB - We present the basic structure of the Cognitive Paradigm Ontology (CogPO) for human behavioral experiments. While the experimental psychology and cognitive neuroscience literature may refer to certain behavioral tasks by name (e.g., the Stroop paradigm or the Sternberg paradigm) or by function (a working memory task, a visual attention task), these paradigms can vary tremendously in the stimuli that are presented to the subject, the response expected from the subject, and the instructions given to the subject. Drawing from the taxonomy developed and used by the BrainMap project ( www.brainmap.org ) for almost two decades to describe key components of published functional imaging results, we have developed an ontology capable of representing certain characteristics of the cognitive paradigms used in the fMRI and PET literature. The Cognitive Paradigm Ontology is being developed to be compliant with the Basic Formal Ontology (BFO), and to harmonize where possible with larger ontologies such as RadLex, NeuroLex, or the Ontology of Biomedical Investigations (OBI). The key components of CogPO include the representation of experimental conditions focused on the stimuli presented, the instructions given, and the responses requested. The use of alternate and even competitive terminologies can often impede scientific discoveries. Categorization of paradigms according to stimulus, response, and instruction has been shown to allow advanced data retrieval techniques by searching for similarities and contrasts across multiple paradigm levels. The goal of CogPO is to develop, evaluate, and distribute a domain ontology of cognitive paradigms for application and use in the functional neuroimaging community. PMID- 21643734 TI - Contrast-enhanced ultrasound: a new method for TIPS follow-up. AB - PURPOSE: To determine the diagnostic value of contrast-enhanced ultrasound (CEUS) for the detection of TIPS (transjugular intrahepatic portosystemic shunt) complications. MATERIALS AND METHOD: 67 cirrhotic patients who underwent TIPS between 2001 and 2008 were retrospectively reviewed. Sixty-two vascular examinations in 37 patients for suspicion of TIPS dysfunction based on the clinical or radiological criteria were analyzed and compared with the 62 related Doppler and CEUS examinations obtained previously. Abnormal CEUS was defined as poor opacification of the prosthesis compared to the native portal vein, stent stenosis, hepatic vein stenosis, and occlusion of the stent. RESULTS: Among the 62 vascular examinations, 56 were considered as pathologic, including: 20 occlusions, 25 stent stenoses, 9 hepatic vein stenoses, 1 arterial-TIPS fistula, and 1 strong flow stealing through a voluminous paraumbilical vein associated with a patent shunt. 50 were exactly correlated with a previous CEUS examination, including 20/20 occlusions (100%), 23/25 stent stenoses (91%), 5/9 hepatic vein stenoses (56%), 1/1 fistula (100%), and 1/1 strong flow stealing (100%). Two CEUS and 14 Doppler examinations were false negative. CONCLUSION: CEUS allows direct visualization of intra-prosthetic flow, with a qualitative and anatomic study, in addition to the Doppler examination. This is a new, simple, and effective technique for TIPS follow-up. PMID- 21643733 TI - Publication bias in neuroimaging research: implications for meta-analyses. AB - Neuroimaging and the neurosciences have made notable advances in sharing activation results through detailed databases, making meta-analysis of the published research faster and easier. However, the effect of publication bias in these fields has not been previously addressed or accounted for in the developed meta-analytic methods. In this article, we examine publication bias in functional magnetic resonance imaging (fMRI) for tasks involving working memory in the frontal lobes (Brodmann Areas 4, 6, 8, 9, 10, 37, 45, 46, and 47). Seventy-four studies were selected from the literature and the effect of publication bias was examined using a number of regression-based techniques. Pearson's r correlation coefficient and Cohen's d effect size estimates were computed for the activation in each study and compared to the study sample size using Egger's regression, Macaskill's regression, and the 'Trim and Fill' method. Evidence for publication bias was identified in this body of literature (p < 0.01 for each test), generally, though was neither task- nor sub-region-dependent. While we focused our analysis on this subgroup of brain mapping studies, we believe our findings generalize to the brain imaging literature as a whole and databases seeking to curate their collective results. While neuroimaging databases of summary effects are of enormous value to the community, the potential publication bias should be considered when performing meta-analyses based on database contents. PMID- 21643735 TI - Overload hepatitides: quanti-qualitative analysis. AB - Diffuse liver diseases have a definitive radiological importance due to the ability of MR imaging to demonstrate abnormalities before the patient is symptomatic or the liver damage is advanced. Biopsy procedures are invasive, may lead to complications and have a sample bias. Imaging biomarkers target to fat, water, and iron tissue concentrations may be considered as hepatic virtual biopsies. There is a need to identify a rapid and practicable method to accurately quantify liver steatosis, differentiate steatohepatitis from simple steatosis, grade the necroinflammatory activity, calculate the liver iron burden and monitor overload progression. MR is used in the evaluation of diffuse liver disorders with accurate approaches such as the use of chemical shift, Dixon vector analysis, turbo spin echo fat suppression, and T2* gradient echo techniques. These methods are influenced by some factors like proportional ambiguity, T1 and T2* effects on signal decay, adding a significant bias in the combined fat-water-iron quantification. A GRE multi-echo chemical shift sequence was configured to independently calculate fat, water, and iron parametric liver images. It is now necessary to conduct a pilot project in order to validate this method in a group of subjects without and with different grades of fat, water, and iron liver changes. PMID- 21643736 TI - Abdominal cocoon with small bowel obstruction: two case reports. AB - Abdominal cocoon is a rare condition that results in an intestinal obstruction due to total or partial encapsulation of the small bowel by a fibrocollagenous membrane. Treatment is surgical resection of the membrane and free the bowel. Preoperative diagnosis is possible with combination of sonography and CT scan. We report two cases where the diagnosis of abdominal cocoon was suggested preoperatively based on the sonography and CT scan of abdomen. PMID- 21643737 TI - Peginterferon alfa-2a is superior to peginterferon alfa-2b in the treatment of naive patients with hepatitis C virus infection: meta-analysis of randomized controlled trials. AB - BACKGROUND: Pegylated interferon (PEGIFN) and ribavirin combination is the standard of care for the treatment of chronic hepatitis C virus (HCV) infection. Studies comparing the efficacy and safety of PEGIFN alfa-2a and PEGIFN alfa-2b in treatment-naive HCV-infected patients have shown conflicting results. AIM: We performed a systematic review and meta-analysis of studies comparing the efficacy and safety of PEGIFN alfa-2a and PEGIFN alfa-2b in HCV-infected patients naive to treatment. METHODS: Nine studies (five abstracts) with 3,546 patients (1,771 treated with PEGIFN alfa-2a) comparing PEGIFN alfa-2a and PEGIFN alfa-2b in treatment-naive HCV patients were analyzed. Efficacy outcomes were sustained virologic response (SVR) and treatment discontinuation rates due to serious adverse effects (SAE). RESULTS: Pooled data on outcomes (reported as odds ratios [ORs] with 95% confidence intervals [CIs]: [OR (95% CI)]) showed higher SVR in patients treated with PEGIFN alfa-2a as compared to treatment with PEGIFN alfa-2b [1.36 (1.07-1.73); P=0.01]. Subgroup analysis of good quality studies on SVR in genotypes 2 and 3 also favored PEGIFN alfa-2a over PEGIFN alfa-2b (1.91 [1.09 3.37]; P=0.02). SVR results obtained with the two types of IFN showed no impact of viral load and the presence or absence of cirrhosis. Treatment discontinuation rates due to SAE, reported in six studies (two abstracts) on 3,211 patients (1,604 treated with PEGIFN alfa-2a), were similar in the two types of PEGIFN [0.66 (0.37-1.16); P=0.15]. CONCLUSIONS: PEGIFN alfa-2a has superior efficacy with higher SVR as compared to PEGIFN alfa-2b in treatment-naive HCV-infected patients. The safety profile of the two types of PEGIFN was similar. PMID- 21643738 TI - Risk of functional gastrointestinal disorders in U.S. military following self reported diarrhea and vomiting during deployment. AB - INTRODUCTION: Military personnel are frequently deployed to regions of the world with high travelers' diarrhea (TD) rates. Pathogens associated with TD have been linked to several post-infectious sequelae, including functional gastrointestinal disorders (FGD), such as irritable bowel syndrome (IBS) and functional dyspepsia. Furthermore, stress associated with deployment may potentiate the increased FGD risk. AIM: We sought to assess whether self-reported diarrhea, vomiting, and stressors during deployment were associated with increased FGD risk. METHODS: Using active duty military medical encounter data from the Defense Medical Surveillance System (DMSS), we conducted a matched case-control study to assess the odds of FGD (IBS, functional constipation, functional diarrhea, dyspepsia) following self-reported diarrhea or vomiting during deployment. Only first-time deployers with detailed self-reporting of deployment-related exposures from 2008 and 2009 were included. Univariate and multivariate analyses were performed. RESULTS: A total of 129 cases of FGD were identified, with the following distribution: constipation (n = 67), dyspepsia (n = 15), IBS (n = 22), and overlapping disorders (n = 25). Diarrhea and/or vomiting during deployment were significantly associated with the development of FGD. Other demographic factors were also associated with variable risk. We found no consistent effect of war related stressors or non-combat-related correlates of stress. CONCLUSIONS: Deployment-related TD is common in deployed military personnel and is associated with an increased risk of several FGD. When considering effective countermeasures and mitigation strategies, both the acute effects and chronic sequelae of enteric infections should be considered. Increased emphasis on existing and novel primary prevention strategies are needed, as well as outcome studies among those developing these conditions. PMID- 21643739 TI - Serum interleukin-33 levels in patients with gastric cancer. AB - BACKGROUND: Interleukin-33 (IL-33) is a novel member of the IL-1 family of cytokines, and it is closely related to IL-18, one of the best characterized members of the IL-1 family. It's been demonstrated that elevated levels of IL-18 are involved in a wide variety of tumors, especially in gastric cancer. AIMS: The purpose of this study was to determine the correlations between serum IL-33 levels and the clinicopathologic features in gastric cancer patients. METHODS: Serum samples were collected from 68 patients with gastric cancer and 57 controls. Serum IL-33 levels were measured by ELISA. Classical tumor markers of CEA and CA19-9 levels were routinely detected by chemiluminescence immunoassay. Western blot analysis was used to detect IL-33 expression in gastric cancer tissue samples and cell lines. The relationship between serum levels of IL-33 and clinical characteristics in patients was analyzed. RESULTS: IL-33 levels in the serum of gastric cancer patients were significantly elevated in comparison with that of healthy volunteers. Furthermore, higher serum levels of IL-33 in gastric cancer patients were found to correlate with several poor prognostic factors like depth of invasion, distant metastasis and advanced stage (stage III/IV). On the other hand, serum IL-33 levels did not correlate with CEA and CA19-9. The expression of IL-33 protein was upregulated in carcinoma tissues in comparison with matched normal tissues, and no statistically significant difference was found between the four gastric cancer cell lines and human gastric epithelial cell line GES-1. CONCLUSIONS: Serum IL-33 may be a useful biomarker for predicting the prognosis of gastric cancer. PMID- 21643740 TI - Laparoscopic duodenojejunostomy for superior mesenteric artery syndrome. PMID- 21643741 TI - Follow-up study of transjugular intrahepatic portosystemic shunt in the treatment of portal hypertension. AB - OBJECTIVE: To evaluate efficacy and complications for cirrhotic patients with variceal bleeding receiving TIPS. METHODS: Of 137 patients who underwent TIPS from 2002 to 2009, 80 patients were included in this study. Information about the incidence of portosystemic encephalopathy (PSE) and rebleeding, and factors then which affected them, was collected by telephone call, letters, or follow-up visits in the out-patient department. RESULTS: (1) TIPS can significantly reduce portal pressure and the risk of variceal bleeding. (2) A coated stent during TIPS can significantly reduce the occurrence of rebleeding in contrast with use of a bare stent (13.51% vs. 32.56%, P < 0.05). (3) Incidence of PSE is related to the diameter of the stent; the wider the stent used, the greater the incidence of PSE. TIPS using the left branch of the portal vein can reduce the incidence of PSE. (4) TIPS combined with embolization has no effect on the incidence of rebleeding or PSE. (5) Mean survival was 77.098 months (95% CI, 68.568-85.628) and median survival 82.000 months (95% CI, 68.539-95.461) according to Kaplan Meier survival analysis. CONCLUSION: It is suggested that coated stents should be used to reduce rebleeding. It is recommended that 8 mm stents should be used for CHILD A and C patients and 8-10 mm stents for CHILD B patients during TIPS, and that TIPS should be considered as first-line therapy because it improves cumulative survival for cirrhotic patients with gastroesophageal variceal bleeding. PMID- 21643742 TI - DNA aptamer beacon assay for C-telopeptide and handheld fluorometer to monitor bone resorption. AB - A novel DNA aptamer beacon is described for quantification of a 26-amino acid C telopeptide (CTx) of human type I bone collagen. One aptamer sequence and its reverse complement dominated the aptamer pool (31.6% of sequenced clones). Secondary structures of these aptamers were examined for potential binding pockets. Three-dimensional computer models which analyzed docking topologies and binding energies were in agreement with empirical fluorescence experiments used to select one candidate loop for beacon assay development. All loop structures from the aptamer finalists were end-labeled with TYE 665 and Iowa Black quencher for comparison of beacon fluorescence levels as a function of CTx concentration. The optimal beacon, designated CTx 2R-2h yielded a low ng/ml limit of detection using a commercially available handheld fluorometer. The CTx aptamer beacon bound full-length 26-amino acid CTx peptide, but not a shorter 8-amino acid segment of CTx peptide which is a common target for commercial CTx ELISA kits. The prototype assay was shown to detect CTx peptide from human urine after creatinine and urea were removed by size-exclusion chromatography to prevent nonspecific denaturing of the aptamer beacon. This work demonstrates the potential of aptamer beacons to be utilized for rapid and sensitive bone health monitoring in a handheld or point of-care format. PMID- 21643743 TI - Effect of indigenous plant extracts on calcium oxalate crystallization having a role in urolithiasis. AB - Crystallization process has a major role in urolithiasis. In the present study, effect of two indigenous plants extracts namely Boerhavia diffusa and Bryophyllum pinnatum extract was determined on the crystallization of calcium oxalate crystals. Effect on the number, size and type of calcium oxalate crystals was observed. Results showed significant activity of both extracts against calcium oxalate crystallization at different concentrations (P < 0.05). Size of the crystals gradually reduced with the increasing concentration of both extracts. The number of calcium oxalate monohydrate crystals which are injurious to epithelial cells gradually reduced and at the highest concentration of extracts (100 mg/ml) completely disappeared (P < 0.05). These results confirm that B. diffusa and B. pinnatum extracts have antiurolithic activity and have the ability to reduce crystal size as well as to promote the formation of calcium oxalate dihydrate (COD) crystals rather than monohydrate (COM) crystals. Control of crystal size and formation of COD rather than COM crystals, in combination with the diuretic action of extracts is an important way to control urolithiasis. PMID- 21643744 TI - Polymorphism of CAG repeats in androgen receptor of carnivores. AB - Androgen effect is mediated by the androgen receptor (AR). The polymorphism of CAG triplet repeat (polyCAG), in the N-terminal transactivation domain of the AR protein, has been involved either in endocrine or neurological disorders in human. We obtained partial sequence of AR exon 1 in 10 carnivore species. In most carnivore species, polyglutamine length polymorphism presented in all three CAG repeat regions of AR, in contrast, only CAG-I site polymorphism presented in primate species, and CAG-I and CAG-III sites polymorphism presented in Canidae. Therefore, studies focusing on disease-associated polymorphism of poly(CAG) in carnivore species AR should investigate all three CAG repeats sites, and should not only consider CAG-I sites as the human disease studies. The trinucleotide repeat length in carnivore AR exon 1 had undergone from expansions to contractions during carnivores evolution, unlike a linear increase in primate species. Furthermore, the polymorphisms of the triplet-repeats in the same tissue (somatic mosaicism) were demonstrated in Moutain weasel, Eurasian lynx, Clouded leopard, Chinese tiger, Black leopard and Leopard AR. And, the abnormal stop codon was found in the exon 1 of three carnivore species AR (Moutain weasel, Eurasian lynx and Black leopard). It seemed to have a high frequency presence of tissue-specific somatic in carnivores AR genes. Thus the in vivo mechanism leading to such highly variable phenotypes of the described mutations, and their impact on these animals, are worthwhile to be further elucidated. PMID- 21643745 TI - Characterization of the first specific Jasmonate biosynthetic pathway gene allene oxide synthase from Artemisia annua. AB - Allene oxide synthase (AOS) is the first committed step in the biosynthetic pathway of Jasmonate. In this study, a full-length cDNA of AOS gene (named as AaAOS) was cloned from Artemisia annua. The gene was 1891 bp in size containing an open reading frame (1581 bp) encoding 526 amino acids. Comparative and bioinformatic analysis revealed that the deduced protein of AaAOS was highly homologous to AOSs from other plant species. Phylogenetic analysis indicated that the protein of AaAOS belonged to the dicotyledonous group, which was consistent with the category of A. annua. Southern blot analysis revealed that it was a low copy gene. Quantitative Real-time PCR (qRT-PCR) analysis showed that AaAOS mRNA accumulated most abundantly in leaves and flowers. The qRT-PCR analysis revealed that MeJA, ABA and ethylene treatments significantly enhanced AaAOS transcript expression. PMID- 21643746 TI - HFE mutations and transferrin C1/C2 polymorphism among Croatian patients with schizophrenia and schizoaffective disorder. AB - The aim of this study was to investigate the possible influence of hemochromatosis gene mutations (HFE-C282Y and H63D) and transferrin gene C2 variant (TF-C2) on susceptibility to schizophrenia and schizoaffective disorder and/or age at first hospital admission. Genotyping was performed in 176 Croatian patients and 171 non-psychiatric Croatian controls using PCR-RFLP analyses. Regarding the H63D mutation, allele and genotype frequencies reached boundary statistical significance. Other allele and genotype distributions were not significantly different between two groups. We also analyzed age at first hospital admission as a continuous variable using the non-parametric Mann-Whitney U-test and Kruskal-Wallis test, and multiple regression analysis. The results of these tests were negative. We concluded that investigated HFE mutations and TF-C2 variant are not high-risk genetic variants for schizophrenia/schizoaffective disorder in our population. Also our data do not support their impact on age at onset of the first psychotic symptoms. PMID- 21643747 TI - In vivo analysis of translation initiation sites in Plasmodium falciparum. AB - Regulation of gene expression in the malaria parasite Plasmodium falciparum is tightly controlled and little is known about the many steps involved. One step i.e. translation initiation is also poorly understood and in P. falciparum, choice of the translation initiation site (TIS) is a critical decision largely due to the high frequency of AUGs in the relatively long 5' untranslated regions of parasite mRNAs. The sequences surrounding the TIS have a major role to play in translation initiation and this report evaluates these sequences by mutational analysis of the heat shock protein 86 gene, transient transfection and reporter assays in the parasite. We find that purines at the -3 and +4 positions are essential for efficient translation in P. falciparum, similar to other eukaryotes. Interestingly, a U at the -1 position results in 2.5-fold higher reporter activity compared to wild type. Certain classes of protein biosynthetic genes show higher frequencies of U at the -1 position, suggesting that these genes may exhibit higher levels of translation. This work defines the optimal sequences for TIS choice and has implications for the design of efficient expression vectors in an important human pathogen. PMID- 21643748 TI - Molecular characterization of CART, AgRP, and MC4R genes and their expression with fasting and re-feeding in common carp (Cyprinus carpio). AB - Cocaine and amphetamine-regulated transcript (CART), agouti-related proteins (AgRP) and Melanocortin 4 Receptor (MC4R) involves in the control of appetite. The genes were cloned and characterized, and their regulation was studied in common carp. The CARTI and CARTII genes encode 117- and 120-amino acids, respectively. The AgRP-1 and AgRP-2 genes encode 128- and 136-amino acids, respectively. CARTI was principally expressed in the brain, eye and ovary, while CARTII was highly expressed in the brain. AgRP-1 was strongly expressed in the brain, intestine, testis and eye, while AgRP-2 was highly expressed only in the gill and eye. The MC4R gene, encoding 326-amino acids, was mainly expressed in the brain testis, pituitary and eye. Phylogenetic analysis had been conducted which implied that both CARTI/CARTII and AgRP-1/AgRP-2 might derived from gene duplication events during genome evolution of common carp. CART, AgRP and MC4R gene expression in brain were decreased after fasting treatment and increased sharply after refeeding comparing with normal fed controls, which suggested that CART, AgRP and MC4R are involved in appetite regulation in common carp. PMID- 21643749 TI - Aptamer inhibits Mycobacterium tuberculosis (H37Rv) invasion of macrophage. AB - There is an urgent need to develop new anti-tuberculosis drugs due to the rising tendency in tuberculosis (TB) around the world. It is known that Mycobacterium tuberculosis (M. tuberculosis) generally infects mammalian host via aerosol route. The pathogenic process has been fully studied that it can initially invade alveolar macrophage, then established stable residence within those phagocytic cells, suggesting that one of the possible ways to prevent this pathogen is to inhibit its invasion and growth in the macrophage. Aptamers from SELEX (Systematic Evolution of Ligands by Exponential Enrichment) have been used to rival virulent M. tuberculosis (H37Rv) in our previous work, and the materials to which aptamers bound were proved to be some outer membrane proteins of H37Rv. In the present study, the interaction between M. tuberculosis and macrophage in the presence of aptamers was investigated in more details. The results suggested that the selective aptamers significantly inhibited H37Rv invasion of macrophage in vitro, and the effect correspond to the binding affinity of these aptamers to H37Rv. The values of equilibrium dissociation constant (Kd) was calculated by flow cytometry, all in the nanomolar range, showed much higher affinity to H37Rv than M. bovis Bacillus Guerin (BCG). Moreover, the aptamer-treated H37Rv can stimulate IFN-gamma, IL-15 and IL-17 secretion of macrophages compared with H37Rv (no treated). In summary, our data indicated that the NK2 aptamer not only acted as anti-tuberculosis agent by inhibiting virulent M. tuberculosis (H37Rv) invasion of macrophage, but also might be used as molecular probe for exploring the interaction between the outer membrane of M. tuberculosis and macrophage. PMID- 21643750 TI - EGY2, a chloroplast membrane metalloprotease, plays a role in hypocotyl elongation in Arabidopsis. AB - Intramembrane proteases control many important processes in a wide variety of organisms through regulated intramembrane proteolysis (RIP). However, very few intramembrane proteases have been characterized in plants. Intriguingly, EGY2 in Arabidopsis belongs to the Site-2 protease (S2P) family that performs RIP. It contains the conserved catalytic motifs, HExxH and NPDG on its multiple transmembrane helices. Four egy2 knockout mutants have significantly shorter hypocotyls and accumulate lower levels of fatty acids in seedlings. Accumulation of fatty acid biosynthesis enzymes in seedlings are also decreased in egy2 knockout mutants. EGY2 protein resides in the chloroplast and EGY2 transcripts are found throughout the plant except root. Recombinant EGY2 protein cleaves beta casein in an ATP-independent manner. These results together suggest that EGY2 metalloprotease plays a role in hypocotyl elongation likely through a RIP dependent process to regulate the coordinated expression of nuclear- and plastid encoded genes. PMID- 21643751 TI - Common BRCA1 and BRCA2 mutations in breast cancer families: a meta-analysis from systematic review. AB - A number of molecular epidemiological studies have been conducted the screening for BRCA1 and BRCA2 mutations in breast cancer patients with a positive family history of breast and/or ovarian cancer and reported many common mutations in BRCA1 and BRCA2 associated in breast cancer in different population and different ethnicity. However, it's still lack of a systematic analysis on these mutations. To comprehensively evaluate the frequency and distribution of common BRCA1 and BRCA2 mutations which associated with breast cancer risk, we address this issue through system review and meta-analysis on 29 relevant published studies by conducting a literature search on PubMed and CNKI. 20 common founder germline mutations were identified from all 29 studies and 4 of BRCA1 (5382insC, 185delAG, 3819del5 and 4153delA) and 2 of BRCA2 (4075delGT, 5802del4) mutations were repeatedly reported twice or more in different articles, respectively. For the BRCA1, after conducting meta-analysis, we found that the overall frequency of 5382insC was 0.09 (95% CI 0.06-0.12), the frequency of 185delAG was 0.07 (95% CI 0.01-0.13), the frequency of 3819del5 was 0.02 (95% CI 0.01-0.04) and the frequency of 4153delA was 0.06 (95% CI 0.03-0.09). For the BRCA2, the overall frequency of 4075delGT was 0.02 (95% CI 0.00-0.03) and the frequency of 5802del4 was 0.07 (95% CI 0.04-0.11). This article provides a set of common mutations for BRCA1 and BRCA2 mutation carriers and the results may help to explore frequencies of BRCA1 and BRCA2 mutations in a given population and will be of significance both for diagnostic testing and for epidemiological studies. PMID- 21643752 TI - Spatial distribution pattern analysis of Dof1 transcription factor in different tissues of three Eleusine coracana genotypes differing in their grain colour, yield and photosynthetic efficiency. AB - In the present study Dof1 gene of finger millet was cloned and sequenced. In silico analysis reveals 61% identity with the Sorghum bicolor and 57% identity with the Oryza sativa Dof1 sequence. A comparative analysis of gene sequences from different crops and three finger millet genotypes {Brown (PRM-1), Golden (PRM-701) and White (PRM-801)} differing in grain colour, yield and photosynthetic efficiency showed a high degree of sequence identity of Dof1 sequence gene ranging from 22 to 70% as evident from distance matrix of the built phylogenetic tree showing two major clusters. A total of five conserved motifs were observed in Dof1 sequences of different cereals. Motif 1 with multilevel consensus sequence CKNCRRYWTKGGAMRNVPVG contains zinc finger Dof domain. Motif 3 and motif 5 contains protein kinase phosphorylation site. Motif 2 contains Dof domain and zinc finger N-glycosylation site while motif 4 is involved in Zinc finger type profiling. Further, we studied the spatial distribution of Dof1 gene in three vegetative tissues (root, stem and flag leaf) as well as four stages of developing spikes (S1, S2, S3 and S4) of the three finger millet genotypes using qualitative and quantitative PCR based approaches. Physiological parameters (plant height, leaf area, chlorophyll content, SPAD value and photosynthetic efficiency) at the time of flowering was found to be highest in white (PRM-801) genotype followed by golden (PRM-701) and brown (PRM-1) genotype. Semi quantitative RT-PCR and quantitative real-time PCR analysis revealed that the expression of Dof1 is highest in leaves and lowest in roots, which suggests its role in regulation of photosynthesis-related genes and carbon skeleton synthesis. Also at grain maturity stage, expression of Dof1 was higher in white (PRM-801) genotype followed by golden (PRM-701) and brown (PRM-1) genotype. The result is suggestive of Dof1 role in the accumulation of grain protein and yield attribute through regulation of key enzymes involved in source to sink relationship during grain filling stage. PMID- 21643753 TI - Three Brassica rapa metallothionein genes are differentially regulated under various stress conditions. AB - The expression profiles of three Brassica rapa metallothionein genes (BrMT 1-3) were determined in 7-day-old seedlings exposed to various exogenous factors including plant hormones, heavy metals and abiotic stresses. BrMT1, BrMT2, and BrMT3 were representatives of MT gene type 1, type 2, and type 3, respectively, according to their cysteine alignment. BrMT2 showed a relatively higher basal expression level compared to BrMT1 and BrMT3 under normal conditions. The BrMT1 transcript was markedly increased by various factors including ethephon, polyethylene glycol and hydrogen peroxide, with no down-regulation evident. On the contrary, BrMT2 expression was down-regulated by abscisic acid, salicylic acid, and methyl jasmonate. Heavy metals did not increase BrMT2 expression. BrMT3 expression was only marginally and non-significantly up- and down-regulated by the stress conditions tested. Promoter regions of BrMT1 and BrMT2 display different cis-acting elements supporting the different responses of both genes against various stresses. The results demonstrate the differential regulation of BrMT1-3 by various plant exogenous factors, and indicate the utility of the BrMT1 promoter as a multiple stress inducible promoter. PMID- 21643754 TI - MTHFR 677C>T and 1298A>C polymorphisms and male infertility risk: a meta analysis. AB - Epidemiological studies have evaluated the association between MTHFR 677C>T and 1298A>C polymorphisms and risk of male infertility. However, the results from the published studies on the association between these two MTHFR polymorphisms and male infertility risk are conflicting. To derive a more precise estimation of association between the MTHFR polymorphisms and risk of male infertility, we performed a meta-analysis. A comprehensive search was conducted to identify all case-control studies of MTHFR polymorphisms and male infertility risk. We used odds ratios (ORs) with 95% confidence intervals (CIs) to assess the strength of the association. Overall, we found that both 677C>T and 1298A>C polymorphisms were not significantly associated with male infertility risk. However, in stratified analysis by ethnicity, we found that the 677C>T polymorphism was significantly associated with the risk of male infertility in Asian population (TT vs. CC: OR = 1.57, 95% CI: 1.05-2.37, P = 0.03; TT vs. TC + CC: OR = 1.40, 95% CI: 1.05-1.86, P = 0.02; TT + TC vs. CC: OR = 1.34, 95% CI: 1.01-1.77, P = 0.04). Although some modest bias could not be eliminated, this meta-analysis suggested that the MTHFR 677T allele might be a low-penetrant risk factor for male infertility, especially in Asian population. PMID- 21643755 TI - Common functional variants of APOA5 and GCKR accumulate gradually in association with triglyceride increase in metabolic syndrome patients. AB - The common functional variants of the apolipoprotein A5 (APOA5) and the glucokinase regulatory protein genes (GCKR) have been shown to associate with increased fasting triglyceride (TG) levels. Albeit the basic association has been extensively investigated in several populations of different origin, less is known about quantitative traits of them. In our study accumulation rates of four APOA5 (T-1131, IVS3 + G476A, T1259C and C56G) and two GCKR (C1337T and rs780094) functional SNPs were analyzed in patients stratified into four TG quartile groups. Randomly selected 325 metabolic syndrome patients were separated into four quartile (q) groups based on the TG levels as follows q1: TG <1.38 mmol/l; q2: 1.38-1.93 mmol/l; q3: 1.94-2.83 mmol/l; and q4: TG >2.83 mmol/l. We observed significant stepwise increase of prevalence rates of minor allele frequencies in the four plasma TG quartiles for three APOA5 SNPs: -1131C (q1: 4.94%; q2: 8.64%; q3: 11.6%; q4: 12.3%), IVS3 + 476A (q1: 4.32%; q2: 7.4%; q3: 10.36%; q4: 11.1%), and 1259C (q1: 4.94%; q2: 7.41%; q3: 10.4%; q4: 11.7%). The haplotype analysis revealed, that the frequency of APOA5*2 haplotype gradually increased in q2, q3 and q4 (q1: 9.87%; q2: 14.8%; q3: 18.3%; q4: 21%). The distribution of the homozygotes of the two analyzed GCKR variants resembled to the APOA5 pattern. Contrary to the hypothetically predictable linear association coming from the current knowledge about the APOA5 and GCKR functions, the findings presented here revealed a unique, TG raise dependent gradual accumulation of the functional variants of in MS patients. Thus, the findings of the current study serve indirect evidence for the existence of rare APOA5 and GCKR haplotypes in metabolic syndrome patients with higher TG levels, which contribute to the complex lipid metabolism alteration in this disease. PMID- 21643756 TI - Single nucleotide polymorphisms of the purinergic 1 receptor are not associated with myocardial infarction in a Latvian population. AB - The purinergic 1 receptor (P2RY1) has been implicated in development of heart disease and in individual pharmacodynamic response to anticoagulant therapies. However, the association of polymorphisms in the P2RY1 gene with myocardial infarction (MI), and its associated conditions, has yet to be reported in the literature. We evaluated seven known SNPs in P2RY1 for association with MI in a Latvian population. Seven independent parameters that are related to MI [body mass index (BMI), type 2 diabetes (T2D), angina pectoris, hypertension, hyperlipidemia, atrial fibrillation and heart failure] were investigated. No significant association with MI was observed for any of the polymorphisms. Those SNPs for which the P value was close to significance were located in coding or promoter regions. Intriguingly, carriers of the minor allele in the P2RY1 gene locus showed a tendency towards higher onset age for MI, suggesting a possible protective effect of these SNPs against MI or their contribution in progression as opposed to onset. Finally, a linkage disequilibrium (LD) plot was generated for these polymorphisms in the Latvian population. The results of this study suggest that the role of P2RY1 in individuals from Latvian population is likely to be principally involved in platelet aggregation and thromboembolic diseases, and not as a significant contributing factor to the global metabolic syndrome. PMID- 21643757 TI - Association between C1431T polymorphism in peroxisome proliferator-activated receptor-gamma gene and coronary artery disease in Chinese Han population. AB - The C1431T polymorphism in peroxisome proliferator-activated receptor-gamma (PPARgamma) has been shown to be associated with diabetes, obesity, and metabolic syndrome. However, it is unclear whether this polymorphism is associated with coronary artery disease (CAD). Therefore, we conducted a hospital-based case control study with 864 CAD patients and 1008 controls to explore the association between the PPARgamma C1431T polymorphism and risk of CAD in Chinese Han population. Subjects with the variant genotypes (CT + TT) had a 39% decreased risk of CAD relative to CC carriers (adjusted odds ratio, 0.61; 95% confidence interval, 0.49-0.76). Our results suggested that the C1431T polymorphism was associated with a higher body mass index in both CAD patients and controls. Moreover, this polymorphism was also found to be associated with a higher HDL cholesterol level and a lower blood glucose level in CAD patients. In stratified analyses, the T allele was significantly associated with reduced risk of CAD in males, subjects with age <62 years, and non-smokers. In conclusion, the PPARgamma C1431T polymorphism is associated with decreased risk of CAD in Chinese Han population. PMID- 21643758 TI - Association of pig UCP3 gene mutations and back fat thickness in the sixth and seventh rib. AB - Accumulated studies have documented extensive links between UCP3 polymorphisms and pig productive traits, and quantitative trait loci linkage results, on the other hand, provided extensive evidences showing that UCP3 was in the core of several QTLs for carcass and meat quality traits. In this research, we screened two substitutions in coding sequence and one 9-base continuous mutated site in 3'UTR of pig UCP3 gene using the reference population of 293 pigs which were F (2) generation of hybrids between Chinese native Jinhua pigs and European Pietrain. The two missense mutations of G1406A in Exon 3 and T3602C in Exon 5 which led to changes in the G150R and M259T, respectively, were digested by SmaI and introduced Tth111I separately for genotype analysis, and the 9-base continuous mutated site in the 3'UTR was analyzed by an AvaI cleavage. As a result, the 9-base continuous mutated site of 3'UTR manifested significantly close association with the backfat thickness at the sixth and seventh rib, but the polymorphisms of G1406A and T3602C were not associated significantly with any of the seven carcass traits. The same results were shown by RT-qPCR and western blotting. These findings inferred that UCP3 probably has tissue-specific effects on pig carcass traits. PMID- 21643759 TI - Quantitative assessment of the effect of ABCA1 R219K polymorphism on the risk of coronary heart disease. AB - In the past decade, a number of case-control studies have been conducted to investigate the relationship between the ATP-binding cassette transporter A1 (ABCA1) R219K polymorphism and coronary heart disease (CHD). However, the results have been inconclusive. The purpose of the present study is to investigate whether this polymorphism confers significant susceptibility to CHD using a meta analysis. PubMed, Embase and CNKI database were searched to get the genetic association studies. Then data were extracted. Pooled odds ratio (OR) and 95% confidence interval (CI) were calculated. Moreover, subgroup and sensitive analysis were performed. In total, 9,437 cases and 16,270 controls were involved in the meta-analysis. The K219 was significantly associated with CHD (OR = 0.80, 95% CI 0.69-0.92, P(Z) = 0.001). However, significant heterogeneity was present. Further subgroup analysis suggested ethnicity explained much heterogeneity. In Asians, K219 showed a strong protective effect and the pooled OR was 0.69 (95% CI 0.55-0.86 P(Z) = 0.0009). While in Caucasians the result was not significant (OR = 0.87, 95% CI 0.73-1.04, P(Z) = 0.12). In conclusion, our results indicate that the ABCA1 R219K polymorphism is a protective factor of CHD in Asians, but not in Caucasians. PMID- 21643760 TI - Polymorphisms of diacylglycerol acyltransferase 2 gene and their relationship with growth traits in goats. AB - Diacylglycerol acyltransferase (DGAT) plays a critical role in the synthesis of triacylglycerol. In this study, PCR-SSCP and DNA sequencing methods were employed to screen the genetic variations of DGAT-2 gene in 299 goats from three breeds (Boer goat, Chinese Xuhuai white goat and Chinese Haimen goat). Three fragments of DGAT-2 gene were investigated, only exon 3 of DGAT-2 gene showed polymorphism. The alignment between nucleotide sequences of NM_205793.2 in GenBank and the sequencing results of three PCR products with different patterns revealed that there was one mutation (A -> G) in exon 3 of DGAT-2 gene, which resulted in amino acid change (Lys -> Arg) and constructed two genotypes (AA, AB). The frequencies of allele A and genotype AA were dominant in all three breeds. And there was no significant difference for genotypic and allelic frequencies among the three breeds. The genotype distributions were in good agreement with Hardy-Weinberg equilibrium (P > 0.05) in each breed. Significant statistical differences were only found in withers heights (P < 0.05) in Xuhuai goat between genotypes. The results indicated that individuals with genotype AA were significantly higher than those of individuals with genotype AB in withers height (P < 0.05). No polymorphism was detected in the intron 3, exon 8 and 3' flanking region. So we suggested that DGAT-2 gene had the close relationship with growth traits in goats. And this mutation could be used as a perfect molecular marker for marker assisted selection (MAS) in animal genetics and breeding. PMID- 21643761 TI - Aging is an organ-specific process: changes in homeostasis of iron and redox proteins in the rat. AB - Organ-specific changes of iron- and redox-related proteins occur with age in the rat. Ferritin, the major iron storage and detoxifying protein, as well as the proteins of the methionine-centered redox cycle (MCRC) were examined in old and young animals, and showed organ-dependent changes. In spleens and livers of aged rats, ferritin (protein) levels were greater than in young ones, and their iron saturation increased, rendering higher ferritin-bound iron (FtBI). Iron saturation of the ferritin molecule in the tongues and sternohyoids of old rats was lower but ferritin level was higher than in young rats, resulting in increased FtBI with age. Ferritin level in the esophagus of older rats was lower than in young rats but its molecular iron content higher thus the total FtBI remained the same. In the larynx, both ferritin and its iron content were the same in young and old animals. MCRC proteins were measured in livers and spleens only. With aging, methionine sulfoxide reductase A and B (MsrA and MsrB) levels in livers and spleens decreased. Thioredoxin1 (Trx) and Trx-reductase1 were elevated in old spleens, but reduced in livers. Aged spleens showed reduced Msr isozyme activity; but in the liver, its activity increased. mRNA changes with age were monitored and found to be organ specific. These organ-specific changes could reflect the different challenges and the selective pathways of each organ and its resultant capacity to cope with aging. PMID- 21643762 TI - Long-term training induces a healthy inflammatory and endocrine emergent biomarker profile in elderly men. AB - The objective of the present study was to investigate the changes in a large panel of emergent geriatric biomarkers in long-term trained elderly men to analyze the effects of long-term exercise on an aged population. We collected blood samples from two groups of male volunteers older than 65 years who maintain a measure of functional independence: one group of sedentary subjects without a history of regular physical activity and the other of subjects who have sustained training, starting during adulthood (mean training time = 49 +/- 8 years). We studied morbidity, polypharmacy, cellular and serological inflammatory parameters, and endocrine mediators. After adjusting for confounding factors, we observed reduced medication intake per subject and lower number of diseases per subject with statistical differences nearly significant in the long-term exercise group. We showed that long-term training was associated with lower levels of white blood cell counts, neutrophil counts, interleukin-6, interleukin-10, interleukin-1 receptor antagonist, and soluble TNF receptor-I. Furthermore, we noted an increase in the concentrations of insulin-like growth factor-1 and dehydroepiandrosterone in the long-term training group. We concluded that long term exercise training from adulthood to old age is clearly associated with a healthy profile of emergent geriatric biomarkers. Long-term training could improve the inflammatory-endocrine imbalance associated with disease, frailty, functional decline, and mortality in elderly men. Our results point to the benefits of prolonged exercise from adulthood to old age. PMID- 21643763 TI - A preliminary, qualitative exploration of the influences associated with drop-out from cognitive-behavioural therapy for problem gambling: an Australian perspective. AB - It has been estimated that 80% of Australians engage in some form of gambling, with approximately 115,000 Australians experiencing severe problems (Productivity Commission 2010). Very few people with problem gambling seek help and, of those who do, large numbers drop-out of therapy before completing their program. To gain insights into these problems, participants who had either completed or withdrawn prematurely from an individual CBT-based problem gambling treatment program were interviewed to examine factors predictive of premature withdrawal from therapy as well as people's 'readiness' for change. The results indicated that there might be some early indicators of risk for early withdrawal. These included: gambling for pleasure or social interaction; non-compliance with homework tasks; gambling as a strategy to avoid personal issues or dysphoric mood; high levels of guilt and shame; and a lack of readiness for change. The study further showed that application of the term 'drop-out' to some clients may be an unnecessarily negative label in that a number appear to have been able to reduce their gambling urges even after a short exposure to therapy. PMID- 21643765 TI - Peripheral immune challenge with viral mimic during early postnatal period robustly enhances anxiety-like behavior in young adult rats. AB - Inflammatory factors associated with immune challenge during early brain development are now firmly implicated in the etiologies of schizophrenia, autism and mood disorders later in life. In rodent models, maternal injections of inflammagens have been used to induce behavioral, anatomical and biochemical changes in offspring that are congruent with those found in human diseases. Here, we studied whether inflammatory challenge during the early postnatal period can also elicit behavioral alterations in adults. At postnatal day 14, rats were intraperitoneally injected with a viral mimic, polyinosinic:polycytidylic acid (PIC). Two months later, these rats displayed remarkably robust and consistent anxiety-like behaviors as evaluated by the open field/defensive-withdrawal test. These results demonstrate that the window of vulnerability to inflammatory challenge in rodents extends into the postnatal period and offers a means to study the early sequelae of events surrounding immune challenge to the developing brain. PMID- 21643764 TI - Behavioral and neurochemical effects of proline. AB - Proline is an amino acid with an essential role for primary metabolism and physiologic functions. Hyperprolinemia results from the deficiency of specific enzymes for proline catabolism, leading to tissue accumulation of this amino acid. Hyperprolinemic patients can present neurological symptoms and brain abnormalities, whose aetiopathogenesis is poorly understood. This review addresses some of the findings obtained, mainly from animal studies, indicating that high proline levels may be associated to neuropathophysiology of some disorders. In this context, it has been suggested that energy metabolism deficit, Na(+),K(+)-ATPase, kinase creatine, oxidative stress, excitotoxicity, lipid content, as well as purinergic and cholinergic systems are involved in the effect of proline on brain damage and spatial memory deficit. The discussion focuses on the relatively low antioxidant defenses of the brain and the vulnerability of neural tissue to reactive species. This offers new perspectives for potential therapeutic strategies for this condition, which may include the early use of appropriate antioxidants as a novel adjuvant therapy, besides the usual treatment based on special diets poor in proline. PMID- 21643766 TI - Characterization of the complete genomic sequence of the rinderpest virus Fusan strain cattle type, which is the most classical isolate in Asia and comparison with its lapinized strain. AB - In this study, we characterized the rinderpest virus (RPV) Fusan strain cattle type (B), which is the most classical isolate in Asia, by complete genomic sequence analysis and compared it with its lapinized Nakamura III (L) strain. The transversion rates of the M, F, and H genes were higher than those of other genes. In contrast, the deduced amino acid (aa) substitution rates of the P, C, and V genes were higher than those of other genes, although their transversion rates were not higher. The characteristic nucleotide (nt) or aa residues of the cattle-virulent B and Kabete 'O' strains were observed in the P/C/V, M, and L genes. According to these results, we speculate that nt/aa substitution in the P/C/V genes is one of the key determinants for the difference in the pathogenicity to cattle of the B and L strains. PMID- 21643767 TI - A novel sapelovirus-like virus isolation from wild boar. AB - A novel sapelovirus-like virus was isolated from a wild boar (Sus scrofa). In this study, partial viral genomic nucleotide sequences were determined using the rapid determination system of viral nucleic acid sequences (RDV) ver. 3.1, which we recently developed for discovering novel viruses. Phylogenetic analysis of VP1 and 3A proteins and their encoding nucleotide sequences of enteroviruses and sapeloviruses indicated that the isolated virus was closely related to porcine sapelovirus. RT-PCR detected viral sequences in six of 48 wild boar fecal samples. PMID- 21643768 TI - A duplex real-time RT-PCR assay for the simultaneous genogroup-specific detection of noroviruses in both clinical and environmental specimens. AB - Norovirus (NoV) is the major etiological agent causing foodborne and waterborne outbreaks worldwide. We developed a novel duplex real-time quantitative RT-PCR assay designed for the simultaneous detection of and discrimination between NoV genogroups GI and GII, by targeting the short junction region between ORF1 and ORF2, with sensitivity and efficiency comparable to those of each simplex RT-PCR assay. This new duplex assay was evaluated against clinical stool (n = 82) and environmental (groundwater or surface water, n = 60) specimens from South Korea, and the results were compared with those of conventional RT-PCR (cRT-PCR) assays. The duplex assay detected more positive samples than did the cRT-PCR for both clinical (74 vs. 71) and, more strikingly, environmental (24 vs. 10) specimens. No cross-reactivity against specimens containing other enteric viruses such as rotavirus, adenovirus, and poliovirus were observed. These results suggest that this newly developed duplex real-time RT-PCR assay can be used for the sensitive and simultaneous genogroup-specific detection of NoV in both clinical and environmental specimens. PMID- 21643769 TI - Genetic diversity and positive selection analysis of classical swine fever virus isolates in south China. AB - Classical swine fever virus (CSFV) causes a highly contagious disease that leads to significant economic losses in the pig industry worldwide. However, there is a paucity of knowledge on the accurate genotyping of CSFV isolates in south China. This study genotyped the E2 gene of 14 CSFV strains isolated during 2008-2010 from domestic pigs in different districts of south China. Phylogenetic analyses revealed that all of the 14 CSFV isolates were clustered into genetic subgroup 1.1. This contrasts with most parts of China, where group 2 isolates are predominant. Furthermore, the positive selection pressures acting on the E(rns) and E2 envelope protein genes of CSFV were assessed and a site-by-site analysis of the dN/dS ratio was performed to identify specific codons that undergo diversification under positive selection. While no significant evidence for positive selection was observed in E(rns), two positively selected sites at amino acid residues 49 and 72 in the E2 encoding region were identified. Our results revealed that a predominance of subgroup 1.1 CSFV isolates is currently circulating in some districts of south China, which appear to be unrelated to the Chinese C-strain vaccine. Moreover, the envelope protein gene, E2, has undergone positive selection in 14 CSFV strains and two positively selected sites have been identified in this study. Understanding the molecular epidemiology and functional importance of these positively selected amino acid positions could help to predict possible changes in virulence, the development of vaccines and disease control. PMID- 21643770 TI - Lung and pleural CT signs predict deaths: 10-year follow-up after lung cancer screening of asbestos-exposed workers. AB - PURPOSE: To work out the predictive value of pathological (HR)CT signs concerning long-term mortality among those screened for lung cancer. METHODS: Five hundred and eighty four construction workers (574 males, 10 females) were originally screened for lung cancer and found negative. Their images were also scored for several lung and pleural signs. Mortality data were checked from the National Registry of Causes of Death. Cox regression adjusted for age, sex, smoking, BMI, and asbestos exposure was used to explore the relations between the radiological signs and deaths. The mean follow-up time was 10.53 years (0.56-12.98 years) and a total of 6,150 person years were followed up. RESULTS: Altogether, 185 deaths occurred (64 cardiovascular, 51 cancer, 24 non-cancer respiratory deaths, and 46 deaths from other causes). All studied emphysema signs were significant predictors of all-cause deaths as were most fibrosis signs (subpleural nodules, septal lines, parenchymal bands, and honeycombing), ground-glass opacities, thickened bronchial walls, pleural plaque extent, and adherences. Cardiovascular deaths were significantly associated with paraseptal emphysema and bullae. Several lung/pleural signs also predicted cancer and respiratory deaths. CONCLUSION: Pathological lung/pleural CT signs found in screening seem to predict deaths in long term, which may require more careful medical surveillance of such individuals. Further studies are needed to generalize the present findings to general population. PMID- 21643771 TI - The persistence of allergen exposure favors pulmonary function decline in workers with allergic occupational asthma. AB - BACKGROUND: In asthmatics, a rapid decline in pulmonary function is observed, likely as a consequence of airways remodeling. Persistence of allergen exposure in patients with occupational asthma (OA) maintains chronic bronchial inflammation, resulting in a more severe lung function decline. Few studies were performed on the effects of allergen exposure cessation. OBJECTIVE: This study aims at evaluating the influence of allergen exposure cessation on respiratory decline in allergic asthmatic workers. METHODS: Two groups of workers with allergic OA were selected. The first group (30 workers) changed job after the diagnosis and was no more exposed to sensitizing allergens, and the second group (28 subjects) did not and, as a consequence of preventive measures in the work place, was exposed to a lower level of allergens. All were treated with conventional therapy, according to GINA protocols. FEV1 changes during a 12-year period were evaluated. RESULTS: Despite pharmacological therapy, the pulmonary function decay slope was steeper in workers continuously exposed to the sensitizing agent (even at reduced level) than in those with a complete cessation of exposure: final FEV1 loss was 512.5 +/- 180 ml versus 332.5 +/- 108 ml, respectively. The difference became significant after 4 years from the cessation of the exposure. CONCLUSIONS: The study shows that the cessation of the exposure to allergen in the work place appears the most effective measure in limiting pulmonary function decline in asthmatic workers and underlines the importance of allergic risk assessment and control in the management of occupational asthma. PMID- 21643772 TI - Low attack rate of novel influenza A (H1N1) virus infection among healthcare workers: a prospective study in a setting with an elaborated containment plan. AB - PURPOSE: This study aimed to determine incidence rates of novel influenza A (H1N1) infection among healthcare personnel with different exposure risks during the 2009 H1N1 pandemic. METHODS: From August 2009 until April 2010, 66 healthcare workers from a 410 bed teaching hospital in Amsterdam were monitored. The following three different exposure groups were created: a high- (n = 26), intermediate- (n = 20), and low-risk group (n = 20). Throat swabs were collected each week and analyzed by real-time reverse transcriptase-polymerase chain reaction (RT-PCR) in order to detect the H1N1 virus. Blood was drawn at study enrollment and once monthly thereafter, and serum specimens were tested with an H1N1-specific hemagglutination-inhibition serologic assay. Influenza-like signs and symptoms were assessed weekly. RESULTS: One of 26 high-risk group participants proved H1N1 positive once by RT-PCR. This corresponds to an incidence rate in the high-risk group of 5.7/1,000 person weeks (95% CI 0 17/1,000). None of the intermediate- and low-risk group participants proved H1N1 positive by RT-PCR. Significant antibody titer rises in convalescent sera were demonstrated in three participants: one was a confirmation of the case that had proved H1N1 positive by RT-PCR; the others occurred in two asymptomatic participants belonging to the low- and high-risk groups. An influenza-like illness was assumed in four participants from the high- (n = 1), intermediate- (n = 1) and low-risk (n = 2) groups; these findings were not confirmed by positive results from either diagnostic test. CONCLUSIONS: This study demonstrates a low incidence rate of influenza A (H1N1) infection among healthcare workers during the 2009 H1N1 pandemic in a setting with high hygiene standards. PMID- 21643773 TI - Solitary extramedullary plasmacytoma of the thyroid involved by papillary carcinoma: a case report and review of the literature. PMID- 21643774 TI - Efficacy and tolerability of a prolonged release ferrous sulphate formulation in iron deficiency anaemia: a non-inferiority controlled trial. AB - BACKGROUND: Iron deficiency anaemia (IDA) is the last stage of iron deficiency, consecutive to an imbalance between iron supply through food intake and iron loss through physiological or pathological processes. As well as by haemoglobin levels, IDA is diagnosed by measuring biomarkers of iron stores. Women are most affected by IDA since their teenage years, as menstruation constitutes a chronic iron loss. Oral supplementation with ferrous sulphate is an effective therapy, but gastrointestinal side effects may impair treatment compliance. METHODS: The present multicentric randomised controlled trial was designed to assess the non inferiority of a ferrous sulphate prolonged release formulation called V0355 with the referential ferrous sulphate Ferrograd(r) in a population of Italian women aged 18-50 years diagnosed for IDA. Three hundred and ninety-nine patients were randomised to receive V0355 (80 mg Fe/day) or Ferrograd(r) (105 mg Fe/day). RESULTS: After 12 weeks of treatment, the difference in the mean haemoglobin level between the two groups was 0.081 g/dL ([-2.986;1.361], p = 0.54), which confirmed the hypothesis of non-inferiority. All the other biochemical parameters (serum iron, serum ferritin, transferrin, and soluble transferrin receptor) and haematological parameters (erythrocytes count, reticulocytes count, haematocrit, and mean corpuscular volume), as well as patient's anaemia-related symptoms, were not different between treatment groups throughout the study. Furthermore, the incidence of gastrointestinal adverse events of moderate and severe intensity was significantly lower (p = 0.007) in the V0355 group (5.6%) than in the Ferrograd(r) group (13.9%). CONCLUSION: V0355 was as efficient as Ferrograd(r) in the treatment of anaemia and exhibited a better gastrointestinal tolerance profile. PMID- 21643775 TI - Protein and peptide probes for molecular imaging. PMID- 21643776 TI - Neurons as ideal change-point detectors. AB - Every computational unit in the brain monitors incoming signals, instant by instant, for meaningful changes in the face of stochastic fluctuation. Recent studies have suggested that even a single neuron can detect changes in noisy signals. In this paper, we demonstrate that a single leaky integrate-and-fire neuron can achieve change-point detection close to that of theoretical optimal, for uniform-rate process, functions even better than a Bayes-optimal algorithm when the underlying rate deviates from a presumed uniform rate process. Given a reasonable number of synaptic connections (order 10(4)) and the rate of the input spike train, the values of the membrane time constant and the threshold found for optimizing change-point detection are close to those seen in biological neurons. These findings imply that biological neurons could act as sophisticated change point detectors. PMID- 21643777 TI - Linear segmental isolation of the left coronary cusp to eliminate ventricular arrhythmia originating in close proximity to left main coronary artery. PMID- 21643778 TI - The prevalence of body contouring surgery after gastric bypass surgery. AB - BACKGROUND: As bariatric surgery has become more popular, more patients are undergoing body contouring surgery after massive weight loss. Many of the surgical procedures performed on the massive weight loss patient are complex and labor-intensive. Therefore, the plastic surgery unit needs to be prepared for a patient's demand. Little literature is available on how frequently patients who have undergone gastric bypass surgery receive body contouring surgery. METHODS: Two hundred fifty-two subjects (out of 425 who were mailed the questionnaire) who had undergone gastric bypass surgery between 2003 and 2009 completed the questionnaire, which obtained information on body image satisfaction and frequency of body contouring surgery after massive weight loss. RESULTS: Of all patients, 74% desire a body contouring surgery after gastric bypass surgery. Fifty-three patients (21%) have undergone a total of 61 body contouring procedures. The most common were abdominoplasties (59%), followed by lower body lifts (20%). In contrast to a positive judgment of the general aspect of the body image satisfaction after massive weight loss, both genders are unsatisfied with body areas like abdomen/waist, breast, and thighs. CONCLUSIONS: Paralleling the increasing use of bariatric surgery, there is a high demand for body contouring surgery. A huge disparity exists between the number of subjects who desire a body contouring surgery and those who actually received it. PMID- 21643779 TI - Gastrobronchial fistula after sleeve gastrectomy and gastric bypass: endoscopic management and prevention. AB - Gastrobronchial fistula (GBF) is a serious complication following bariatric surgery, whose treatment by thoracotomy and/or laparotomy involves a high morbidity rate. We present the outcomes of endoscopic management for GBF as a helpful technique for its healing process. This is a multicenter retrospective study of 15 patients who underwent gastric bypass (n = 10) and sleeve gastrectomy (n = 5) and presented GBF postoperatively (mean of 6.7 months). Ten patients developed lung abscess and were treated by antibiotic therapy (n = 10) and thoracotomy (n = 3). Abdominal reoperation was performed in nine patients for abscess drainage (n = 9) and/or ring removal (n = 4) and/or nutritional access (n = 6). The source of the GBF was at the angle of His (n = 14). Furthermore, 14 patients presented a narrowing of the gastric pouch treated by 20 or 30 mm aggressive balloon dilation (n = 11), stricturotomy or septoplasty (n = 10) and/or stent (n = 7). Fibrin glue was used in one patient. We performed, on average, 4.5 endoscopic sessions per patient. Endotherapy led to a 93.3% (14 out of 15) success rate in GBF closure with an average healing time of 4.4 months (range, 1-10 months), being shorter in the stent group (2.5 * 9.5 months). There was no recurrence during the average 27.3-month follow-up. A patient persisted with GBF, despite the fibrin glue application, and decided to discontinue it. GBF is a highly morbid complication, which usually arises late in the postoperative period. Endotherapy through different strategies is a highly effective therapeutic option and should be implemented early in order to shorten leakage healing time. PMID- 21643780 TI - Comparative clinical outcomes between pediatric and young adult dialysis patients. AB - Published data on the comparative achievement of The Kidney Disease Dialysis Outcome Quality Initiative (KDOQI) recommended clinical performance targets between children and young adults on dialysis are scarce. To characterize the achievement of KDOQI targets among children (<18 years) and young adults (18-24 years) with prevalent end stage renal disease (ESRD), we performed a cross sectional analysis of data collected by the Mid-Atlantic Renal Coalition, in conjunction with the 2007 and 2008 ESRD Clinical Performance Measures Projects. Data on all enrolled pediatric dialysis patients, categorized into three age groups (0-8, 9-12, 13-17 years), and on a random sample of 5% of patients >= 18 years in ESRD Network 5 were examined for two study periods: hemodialysis (HD) data were collected from October to December 2006 and from October to December 2007 and peritoneal dialysis (PD) data were collected from October 2006 to March 2007 and from October 2007 to March 2008. In total, 114 unique patients were enrolled the study, of whom 41.2% (47/114) were on HD and 58.8% (67/114) on PD. Compared to the pediatric patients, young adults were less likely to achieve the KDOQI recommended serum phosphorus levels and serum calcium * phosphorus product values, with less than one-quarter demonstrating values at or below each goal. Multivariate analysis revealed that both young adults and 13- to 17-year-olds were less likely to achieve target values for phosphorus [young adults: odds ratio (OR) 0.04, 95% confidence interval (95% CI) 0.01-0.19, p < 0.001; 13- to 17 year-olds: OR 0.17, 95% CI 0.04-0.77, p = 0.02] and calcium * phosphorus product (young adults: OR 0.01, 95% CI 0.002-0.09, p < 0.001; 13- to 17-year-olds: OR 0.09, 95% CI 0.02-0.56, p = 0.01) than younger children. In summary, there are significant differences in clinical indices between pediatric and young adult ESRD patients. PMID- 21643781 TI - Renal function, renal volume, and blood pressure in infants with antecedent of antenatal steroids. AB - Steroids have been used for more than 20 years in preterm infants to induce pulmonary maturity; however, some long-term effects have been reported, such as insulin resistance and elevation of blood pressure. The aim of our study was to compare renal volume, renal function, and blood pressure in infants between 12-36 months of age with and without antecedent of antenatal steroid treatment. This was a cross-sectional study comprised of three groups of infants (n = 30, respectively): preterm infants with and without antecedent of receiving antenatal steroids, respectively, and full-term infants. Blood pressure, renal volume, glomerular filtration rate, and tubular function were measured. Blood pressure and cystatin C levels and glomerular filtration rate were higher in both groups of preterm infants than in the control group (p < 0.01). However, no difference in any of the tested variables between the steroid and non-steroid group of preterm infants. Renal volume was similar in preterm and control infants. Based on these results, we conclude that prematurity independent of antenatal steroid use is associated with higher cystatin C and blood pressure levels and a higher glomerular filtration rate in infants between 12-36 months of age. PMID- 21643782 TI - Cardiovascular complications in children with HIV infection. AB - Before effective antiretroviral treatment was introduced in children with HIV, cardiovascular complications were a major cause of morbidity and mortality. Now that antiretroviral treatment is the standard of care in this population, there has been a shift from fulminant cardiovascular complications to recognition of cardiovascular issues related to HIV as a chronic disease state and its treatment. This review explores the recent advances in understanding the roles of atherosclerosis, inflammation, biomarkers, and metabolic abnormalities in cardiac function and cardiovascular disease risk in children with HIV. The true risk and prevalence of cardiovascular outcomes in HIV-infected children and adolescents has yet to be realized. Recently, however, investigators are exploring new cardiovascular health challenges that face children living with HIV, and developing strategies to lower the cardiovascular disease risk for these children. PMID- 21643783 TI - Renal issues in HIV infection. AB - Kidney disease remains a prominent complication of HIV disease, despite beneficial effects of antiretroviral therapy on the natural history of HIV associated nephropathy, and on kidney function in general populations of HIV infected patients. Persons of African descent continue to bear a disproportionate burden of severe kidney disease, as is true for the general population. Recently identified genetic variants in the apolipoprotein L1 gene may contribute to this burden. As is also true for the general population, markers of kidney disease, including microalbuminuria, are sensitive predictors of cardiovascular disease and mortality among persons living with HIV. The emerging experience with kidney transplantation also suggests this to be a viable option in selected patients. PMID- 21643784 TI - Cost-utility analysis of a pharmacy-led self-management programme for patients with COPD. AB - OBJECTIVE: To undertake a cost-utility analysis (CUA) of a pharmacy-led self management programme for Chronic Obstructive Pulmonary Disease (COPD). SETTING: A single outpatient COPD clinic at the Mater Hospital, Belfast, Northern Ireland between. METHOD: CUA alongside a randomised control trial. The economic analysis used data from 127 COPD patients aged over 45 years, with an FEV1 of 30-80% of the predicted normal value. Participants received either a pharmacy-led education and self-management programme, or usual care. One year costs were estimated from the perspective of the National Health Service and Personal Social Services and quality-adjusted life years (QALYs) were calculated based on responses to the EQ 5D at baseline, 6 and 12 months. MAIN OUTCOME MEASURE: Cost per QALY gained. RESULTS: The mean differences in costs and effects between the self-management and education programme and usual care were -L671.59 (95 CI%: -L1,584.73 to L68.14) and 0.065 (95% CI; 0.000-0.128). Thus the intervention was the dominant strategy as it was both less costly and more effective than usual care. The probability of the intervention being cost-effective was 95% at a threshold of L20,000/QALY gained. Sensitivity analyses indicated that conclusions were robust to variations in most of the key parameters. CONCLUSION: The self-management and education programme was found to be highly cost-effective compared to usual care. Further research is required to establish what aspects of self-management and education programmes have the greatest impact on cost-effectiveness. PMID- 21643786 TI - Stroke volume variation obtained with Vigileo/FloTracTM system during bleeding and fluid overload in dogs. AB - PURPOSE: Stroke volume variation (SVV) is a parameter for estimating fluid responsiveness. Recently, the VigileoTM and the Flo-TracTM sensor (Edwards Lifesciences, Irvine, CA, USA) were made available for clinical use to estimate SVV. The aim of this study was to investigate the relationship between the circulating blood volume and SVV, measured by the Vigileo-FloTracTM system (SVV FloTrac) or by central venous pressure (CVP), during a dynamic change in circulating blood transfusion volume, using a continuous constant bleeding and fluid-overload model in dogs. METHODS: Ten anesthetized and mechanically ventilated beagles were used. SVV-FloTrac and CVP were measured during a bleeding period (2 ml/kg/min, 15 min), a stabilization period (15 min), a blood transfusion period (2 ml/kg/min, 15 min), and a 6% hydroxyethyl starch solution overload period (2 ml/kg/min, 15 min). RESULTS: SVV-FloTrac changed significantly when more than 8 ml/kg blood was withdrawn or when more than 8 ml/kg blood was transfused. The change in SVV-FloTrac directly reflected the circulating blood volume change during continuous bleeding and blood transfusion. CVP decreased significantly when more than 4 ml/kg blood was withdrawn or when more than 10 ml/kg was infused, and this indicated that the CVP change did not directly reflect the level of the circulating blood volume change. During the stable circulating blood volume period after blood withdrawal, SVV-FloTrac changed significantly but CVP remained constant. During the fluid overload period, CVP, but not SVV-FloTrac, changed significantly. CONCLUSION: SVV-FloTrac is a sensitive indicator of the dynamic circulating blood volume change during both bleeding and transfusion, but not during either the stable circulating blood volume period after blood withdrawal or the fluid-overload period, in mechanically ventilated dogs. PMID- 21643787 TI - Identification and characterization of a natural inter-genotypic (2b/1b) recombinant hepatitis C virus in Japan. AB - A hepatitis C virus (HCV) strain (HC10-0804) recovered from a 12-year-old Japanese female with chronic hepatitis C segregated into discordant genotypes, 2b and 1b, in the 5'UTR/core and NS5B regions, respectively, thus suggesting an inter-genotypic recombination. The HC10-0804 isolate had a genomic length of 9,423 nucleotides (nt), excluding the poly(U) tract at the 3' terminus, and encoded a single open reading frame (ORF) for a polyprotein of 3,014 amino acids (aa). Based on Simplot and Bootscan analyses, the crossover point from 2b to 1b was estimated at nt 3443/3444 (aa 1034/1035), just after the beginning of the NS3 region. Comparison of the entire genomic sequence showed that the HC10-0804 strain was only 90.2% identical to the previously reported 2b/1b recombinant strain (SE-03-07-1689) from the Philippines, whose putative crossover point was 24 nt downstream of that of HC10-0804. These results indicate the circulation of a novel inter-genotypic (2b/1b) recombinant HCV in Japan. PMID- 21643788 TI - Epidemiological and clinical characteristics of pediatric gastroenteritis associated with new viral agents. AB - A 22-month study (2008-2009) was carried out on 273 patients (average age 40 months), admitted with gastroenteritis to the Pediatric Unit of L. Sacco University Hospital in Milan, Italy. Fecal samples were investigated for rotavirus (HRV), norovirus (NoV), adenovirus (AdV), sapovirus (SaV), enterovirus, astrovirus and bocavirus (HBoV). A 38.3% incidence of infection was observed for HRV, followed by NoV (16.2%), HBoV (13.6%), AdV (2.6%) and SaV (0.6%). Clinical evaluation of 109 gastroenteritis patients with confirmed diagnosis was graded by the Ruska-Vesikari scoring system, showing vomiting (78%), diarrhea (96%) and fever (80%). A total of 25 NoV-positive samples were selected for nucleotide sequence analysis. The severity of AdV-associated infection was lower than for NoV, HRV and HBoV. These latter viruses caused similar symptoms that were indistinguishable using clinical information. NoV, HRV and HBoV were often present as mixed infections (13.1%). Sequencing of NoV-positive samples allowed identification of GII.2, GII.3 and GII.4 2006 variants. PMID- 21643789 TI - Anatomical basis of digital rectal examination. AB - PURPOSE: Rectal examination is difficult to carry out by students because of their lack of knowledge and fear. It is therefore necessary to search for methods in order to facilitate its practice. This work mainly focuses on the palpation of the posterior lateral area of the rectum. METHODS: This work bases itself on the study of the average length of indexes and on the anatomical study of the dissection and prints of two pelvises. In the lithotomy position, we can identify three successive levels of exploration of the posterior and lateral area of the rectum. These three levels are defined by the extremity of the index, and the distal and proximal interphalangeal articulations placed successively on the tip of the coccyx. A 180 degrees rotation of the hand enables at each level to identify the parietal structures that the pad of the index comes across, but excludes the palpation of genital organs and rectum. RESULTS: The first level corresponds to the higher part of the anal canal, the ischioanal fossa and the ischium. The second level corresponds to the levator ani muscle, the ischioanal fossa and the pudendal canal. The third level corresponds to the sacrospinous ligament, the ischiatic spine and the internal obturator muscle. CONCLUSIONS: In spite of the significant differences between the lengths of the indexes, the use of these landmarks will facilitate the identification of parietal anatomical structures. The internal organs' palpation will depend on the patient's position, his efforts in pushing, the length of the index, and the way the examiner presses on the perineum. PMID- 21643790 TI - Towards a systemic metabolic signature of the arbuscular mycorrhizal interaction. AB - Our experiments addressed systemic metabolic effects in above-ground plant tissue as part of the plant's response to the arbuscular mycorrhizal (AM) interaction. Due to the physiology of this interaction, we expected effects in the areas of plant mineral nutrition, carbon allocation and stress-related metabolism, but also a notable dependence of respective metabolic changes on environmental conditions and on plant developmental programs. To assess these issues, we analyzed metabolite profiles from mycorrhizal and non-mycorrhizal Lotus japonicus grown under greenhouse conditions at three different time points in the growing season in three different above-ground organs (flowers, sink leaves and source leaves). Statistical analysis of our data revealed a number of significant changes in individual experiments with little overlap between these experiments, indicating the expected impact of external conditions on the plant's response to AM colonization. Partial least square-discriminant analysis (PLS-DA) nevertheless revealed considerable similarities between the datasets, and loading analysis of the component separating mycorrhizal and non-mycorrhizal plants allowed the defining of a core set of metabolites responsible for this separation. This core set was observed in experiments with and without mycorrhiza-induced growth effects. It corroborated trends already indicated by the significant changes from individual experiments and suggested a negative systemic impact of AM colonization on central catabolic metabolism as well as on amino acid metabolism. In addition, metabolic signals for an increase in stress experienced by plant tissue were recorded in flowers and source leaves. PMID- 21643791 TI - The role of memory-related gene polymorphisms, KIBRA and CLSTN2, on replicate memory assessment in the elderly. AB - The role of the CLSTN2 (rs6439886) and KIBRA (rs17070145) SNPs in cognitive impairment was analysed in a 75-76 years old group. Various memory assessment tests were carried out on individuals at baseline and during follow-up investigations, and biallelic genotyping was performed. No influence of the allele status of either SNPs was observed on any memory test. No increased risk of any type of late development, and cognitive impairment was associated with rs6439886 or rs17070145. PMID- 21643793 TI - An improved approach to steady-state analysis of monoamine oxidases. AB - The search for new monoamine oxidase inhibitors aims to identify potential lead compounds that are more potent and selective than current drugs for use in treating a variety of neuropsychiatric and neurodegenerative conditions. An integral part of this process is a kinetic examination of monoamine oxidases in the presence of the inhibitor, to determine potency and selectivity and to obtain information on mechanism. To date, kinetic data obtained with a probe substrate have been analysed by fitting to the Michaelis-Menten equation which describes a unireactant process in which velocity is related to substrate concentration in a rectangular hyperbolic manner. In this study, we present evidence that monoamine oxidase activity is often not adequately described by this approach. We outline a novel equation strategy that takes account of substrate and inhibitor binding to oxidised and reduced enzyme forms, and quantifies differences between substrates and inhibitors in this regard. When combined with plate reader-based experimental techniques that allow large numbers of substrate and inhibitor concentrations to be used, and the global nonlinear regression facilities of GraphPad Prism software, this straightforward approach allows more appropriate analyses of monoamine oxidase by non-experts than has previously been possible. PMID- 21643792 TI - Kynurenines and headache. AB - In parallel to serotonin synthesis, the major route of tryptophan catabolism is the kynurenine pathway, which produces neuroactive metabolites. Among these substances, kynurenic acid has potential neuroprotective action blocking glutamate release and glutamatergic neurotransmission. Glutamate is a key player in migraine pathogenesis; it is crucial in the communication of first and second order neurons, and it has an important role in the genesis of cortical spreading depression, which is the electrophysiological correlate for migraine aura and may be involved in the activation of the trigeminal system. Thus, kynurenines may affect the pathogenesis directly, by acting on glutamate receptors and exerting other neuromodulatory effects, and indirectly via an altered serotonin metabolism. This work summarizes our current results regarding the role of the kynurenine system in trigeminal activation and other events occurring during migraine headache. PMID- 21643794 TI - Pelvic hemophilic pseudotumor presenting as severe sciatic pain in a patient with no history of hemophilic symptoms. PMID- 21643795 TI - Dangerous cervical radiculopathy by Lemierre's syndrome. PMID- 21643796 TI - Single-stage treatment of lumbar fracture-dislocation using a combined anterior and posterior approach. PMID- 21643797 TI - Autozygosity mapping of a large consanguineous Pakistani family reveals a novel non-syndromic autosomal recessive mental retardation locus on 11p15-tel. AB - Autosomal recessive inherited mental retardation is an extremely heterogeneous disease and accounts for approximately 25% of all non-syndromic mental retardation cases. Autozygosity mapping of a large consanguineous Pakistani family revealed a novel locus for non-syndromic autosomal recessive mental retardation (NS-ARMR). The affected individuals showed low IQ and cognitive impairment without any neurological, skeletal, and biochemical abnormalities. All known NS-ARMR genes were excluded by STS markers, so autozygosity mapping by microarray single-nucleotide polymorphism (SNP) analysis were done in all sampled individuals in the family. The nuclear central loop in the five generation family showed homozygosity for a 6-Mb telomeric region on 11p15, whereas all other linkage regions were excluded by calculation of logarithm of odds (LOD) for the SNP microarray data. A maximum LOD score of Z = 3.31 was calculated for the mapped region. These results suggest a novel genetic locus, MRT17, for NS-ARMR. PMID- 21643798 TI - A novel mitofusin 2 mutation causes canine fetal-onset neuroaxonal dystrophy. AB - We recently reported autosomal recessive fetal-onset neuroaxonal dystrophy (FNAD) in a large family of dogs that is not caused by mutation in the PLA2G6 locus (Fyfe et al., J Comp Neurol 518:3771-3784, 2010). Here, we report a genome-wide linkage analysis using 333 microsatellite markers to map canine FNAD to the telomeric end of chromosome 2. The interval of zero recombination was refined by single-nucleotide polymorphism (SNP) haplotype analysis to ~200 kb, and the included genes were sequenced. We found a homozygous 3-nucleotide deletion in exon 14 of mitofusin 2 (MFN2), predicting loss of a glutamate residue at position 539 in the protein of affected dogs. RT-PCR demonstrated near normal expression of the mutant mRNA, but MFN2 expression was undetectable to very low on western blots of affected dog brainstem, cerebrum, kidney, and cultured fibroblasts and by immunohistochemistry on brainstem sections. MFN2 is a multifunctional, membrane-bound GTPase of mitochondria and endoplasmic reticulum most commonly associated with human Charcot-Marie-Tooth disease type 2A2. The canine disorder extends the range of MFN2-associated phenotypes and suggests MFN2 as a candidate gene for rare cases of human FNAD. PMID- 21643799 TI - Metal-on-metal or metal-on-polyethylene for total hip arthroplasty: a meta analysis of prospective randomized studies. AB - BACKGROUND: There has been recent concern regarding the increased use of metal-on metal total hip arthroplasty (MOM-THA) as an alternative to contemporary metal-on polyethylene total hip arthroplasty (MOP-THA), and the choice remains controversial. We performed a meta-analysis to evaluate and compare metal ion concentrations, complications, reoperation rates, clinical outcomes and radiographic outcomes of MOM-THA and MOP-THA. METHODS: We performed a systematic review of English and non-English articles identified from MEDLINE, Embase, the Cochrane Central Register of Controlled Trials, PreMEDLINE and HealthSTAR. Metal ion concentrations, complications, reoperation rates and other outcomes of MOM bearings were compared with MOP bearings in THA based on relative risks, mean differences and standardized mean difference statistics. RESULTS: Eight prospective randomized trials were identified from 1,075 citations. Our results demonstrated significantly elevated erythrocyte, serum and urine levels of metal ions (cobalt and chromium) among patients who received MOM-THA. No significant differences in titanium concentrations or total complication or reoperation rates were found between MOM-THA and MOP-THA. Clinical function scores and radiographic evaluations were similar between the two groups. CONCLUSIONS: This analysis found insufficient evidence to identify any clinical advantage of MOM-THA compared with MOP-THA. Although cobalt and chromium concentrations were elevated after MOM-THA, there were no significant differences in total complication rates (including all case mortality) between the two groups in the short- to mid-term follow-up period. The MOM bearing option for THA should be used with caution. PMID- 21643800 TI - Minimum 5-year results of focal articular prosthetic resurfacing for the treatment of full-thickness articular cartilage defects in the knee. AB - INTRODUCTION: The purpose of this study was to evaluate the results of a contoured focal articular femoral condyle resurfacing prosthetic in the treatment of full-thickness cartilage and osteochondral defects at the medial femoral condyle of the knee beyond 5 years. METHODS: In a multicenter case series, preoperative and follow-up scores of the Knee Injury and Osteoarthritis Outcome Score (KOOS), SF-36 and Tegner activity scale were evaluated. Standard radiographs were performed to evaluate the progression of osteoarthritis. Patients were also asked to report their satisfaction. RESULTS: A total of 21 patients were included in this study. The average follow-up was 5.3 years. The average age at the time of resurfacing was 54 years. Average KOOS scores significantly (P <= 0.005) improved for pain (51.1 to 77.6), symptoms (57.9 to 79.5), activities of daily living (ADL) (58.8 to 82.4), sports (26.3 to 57.8) and quality of life (QOL) (34.4 to 55.0). The Tegner activity level improved significantly (P <= 0.02) from 2.9 to 4. The physical health value of the SF-36 increased by 15.2 to 46.9 compared to the preoperative value. The mental health value almost (51.2) remained unchanged. As many as 16/21 of the patients in this series were satisfied with their outcome and would have the operation again. Radiographic results demonstrated solid fixation, preservation of joint space and no change in the osteoarthritic stage. CONCLUSIONS: The device appears to be an effective reconstructive treatment option for large full-thickness cartilage and osteochondral lesions of the knee in middle-aged patients. PMID- 21643801 TI - Theoretical study of crown ethers with incorporated azobenzene moiety. AB - A series of crown ethers containing the azobenzene moiety incorporated into crowns of various sizes [Cr(O(6))(,) Cr(O(7)) and Cr(O(8))] and their corresponding alkali metal cation (Li(+), Na(+), K(+), Rb(+)) complexes have been studied theoretically. The density functional theory (DFT) method was employed to elucidate the stereochemical structural natures and thermodynamic properties of all of the target molecules at the B3LYP/6-31 G(d) and LANL2DZ level for the cation Rb(+). The fully optimized geometries had real frequencies, thus indicating their minimum-energy status. In addition, the bond lengths between the metal cation and oxygen atoms, atomic torsion angles and thermodynamic energies for complexes were studied. Natural bond orbital (NBO) analysis was used to explore the origin of the internal forces and the intermolecular interactions for the metal complexes. The calculated results show that the most significant interaction is that between the lone pair electrons of electron-donating oxygens in the cis-forms of azobenzene crown ethers (cis-ACEs) and the LP (1-center valence antibond lone pair) orbitals of the alkali-metal cations (Li(+), Na(+), K(+) and Rb(+)). The electronic spectra for the cis-ACEs [cis-Cr(O(6)), cis Cr(O(7)) and cis-Cr(O(8))] are obtained by the time-dependent density functional theory (TDDFT) at the B3LYP/6-31 G(d) level. The spectra of the cis-isomers show broad pi -> pi (S(0) -> S(2)) absorption bands at 310-340 nm but weaker n -> pi (S(0) -> S(1)) bands at 480-490 nm. The calculated results are in good agreement with the experimental results. PMID- 21643802 TI - Smoking onset and the time-varying effects of self-efficacy, environmental smoking, and smoking-specific parenting by using discrete-time survival analysis. AB - This study examined the timing of smoking onset during mid- or late adolescence and the time-varying effects of refusal self-efficacy, parental and sibling smoking behavior, smoking behavior of friends and best friend, and parental smoking-specific communication. We used data from five annual waves of the 'Family and Health' project. In total, 428 adolescents and their parents participated at baseline. Only never smokers were included at baseline (n = 272). A life table and Kaplan-Meier survival curve showed that 51% of all adolescents who did not smoke at baseline did not start smoking within 4 years. The risk for smoking onset during mid- or late adolescence is rather stable (hazard ratio between 16 and 19). Discrete-time survival analyses revealed that low refusal self-efficacy, high frequency of communication, and sibling smoking were associated with smoking onset one year later. No interaction effects were found. Conclusively, the findings revealed that refusal self-efficacy is an important predictor of smoking onset during mid- or late adolescence and is independent of smoking-specific communication and smoking behavior of parents, siblings, and (best) friend(s). Findings emphasize the importance of family prevention programs focusing on self-efficacy skills. PMID- 21643803 TI - A case of anaplastic clear-cell ependymoma presenting with high erythropoietin concentration and 1p/19q deletions. AB - We describe a 19-year-old woman with onset of epileptic seizure, and a small mural nodule and multicystic lesions with severe brain edema located in the right frontal lobe. At surgery, the tumor and a clear margin was removed, and symptoms improved postoperatively. Extended local radiotherapy (60 Gy) was performed. Histopathological examination revealed oligodendroglioma-like tumor cells with a perinuclear halo. The tumor cells extended processes toward CD34-positive proliferating vessels, which resemble a basement membrane. These proliferating vessels formed a tumor membrane so that there was a clear margin between the tumor and brain tissue. Tumor cells were positive for epithelial membrane antigen in a dot-like pattern. MIB-1 staining index was 50.6%. Electron microscopy showed cilia and zipper-like junctions, and anaplastic clear-cell ependymoma grade III was diagnosed. A characteristic of the case was formation of a tumor membrane by proliferating tumor blood vessels. Fluorescence in situ hybridization showed 1p/19q deletions, and the concentration of erythropoietin in the cyst fluid was abnormally high, at 1,859.4 mIU/ml. Erythropoietin and erythropoietin receptors were verified with immunohistochemical staining. PMID- 21643804 TI - Cerebellopontine angle endodermal cyst presenting with hemifacial spasm. AB - Intracranial endodermal cysts presenting with hemifacial spasm (HFS) are extremely rare. We report a right cerebellopontine angle (CPA) endodermal cyst in a 56-year-old man who presented with a 6-month history of right-sided hemifacial spasm. Computed tomography revealed a homogenous, well-demarcated, hyperdense lesion extending from prepontine cistern to right CPA. Magnetic resonance imaging demonstrated a right CPA extra-axial cystic lesion protruding into Meckel's cave, with compression of cranial nerves VII and VIII. Light brown, creamy cystic content was totally removed, and the thin cyst wall surrounding cranial nerves VII and VIII and the right vertebral artery was subtotally removed through the right lateral suboccipital approach. On microscopic examination, the cyst wall was composed of mono- to multilayered stratified epithelia, which were lined by ciliated or nonciliated cuboidal cells, with cilia showing the characteristic 9 + 2 pattern. Immunohistochemistry showed positive staining of cells composing the cyst wall with carcinoembryonic antigen, epithelial membrane antigen, cytokeratin 8, and negative staining with cytokeratin 20, and S-100, thereby characterizing endodermal cyst. Postoperatively, the patient was free of facial spasm. PMID- 21643805 TI - Histopathologic and immunohistochemical profile of spinal glioblastoma: a study of six cases. AB - Spinal intramedullary glioblastomas are uncommon tumors and are known to have a very poor prognosis. Only a few studies in the literature have described their histopathological characteristics. We describe the detailed histopathological and immunohistochemical profiles of six cases of spinal glioblastoma. Most of the tumors were located in the cervical or cervicothoracic region. The majority of the patients were young adults (mean age 34.8 years), presenting with a short duration of symptoms of 2 months or less. Their histopathological features were similar to cerebral glioblastoma. Diverse vascular changes like microvascular proliferation, sprouting angiogenesis, sclerosed and thrombosed vessels, along with field necrosis were prominent findings. All tumors were positive for GFAP and negative for EMA. The MIB-1 labeling index was very high (mean 16.7 +/- 3.2%). Five out of six tumors were immunoreactive for p53 protein, and only two showed over-expression of EGFR protein. The predominant expression of p53 in these young patients suggests that spinal glioblastomas are similar to secondary glioblastoma in the cerebral hemispheres, despite the short duration of symptoms in them and vascular changes that are similar to those noted in primary glioblastoma. These observations support the fact that spinal glioblastomas are heterogeneous tumors underlined by complex molecular pathways. Nevertheless, inactivation of the p53 tumor suppressor pathway could play a major role in the genesis of these neoplasms. PMID- 21643806 TI - 18F-FDG uptake in primary lung cancer as a predictor of intratumoral vessel invasion. AB - OBJECTIVE: This study investigated how fluorodeoxyglucose (FDG) uptake on PET in the primary tumor may predict intratumoral vessel invasion (IVI) in it. METHODS: A total of 512 patients with lung neoplasms determined by a surgical procedure and histopathological diagnosis had undergone FDG-PET scanning. RESULTS: Among the 440 cases confirmed to be malignant, the maximum standardized uptake value (SUV(max)) was significantly lower in IVI-negative cases than IVI-positive cases (P < 0.001). In the substudy on adenocarcinoma (AC), SUV(max) was significantly lower in IVI-negative cases too (P < 0.001), but SUV(max) in squamous cell carcinoma was without significant difference. In addition, IVI was associated with a significantly higher probability of lymph node metastasis (P < 0.001). CONCLUSIONS: This study indicates that a malignant lung tumor with higher SUV(max) has a significantly higher probability of IVI and lymph node metastasis, particularly if the malignancy is an AC. PMID- 21643807 TI - Clinical implications of the body surface area method versus partition model dosimetry for yttrium-90 radioembolization using resin microspheres: a technical review. AB - Yttrium-90 (Y-90) radioembolization is becoming established as an effective therapeutic modality for inoperable liver tumors. For resin microspheres, the 'body surface area (BSA)' method and the partition model can both be used for Y 90 activity calculation. The BSA method is semi-empirical, but more commonly used due its simplicity. The partition model is more accurate, scientifically sound and personalized, but less popular due to its complexity. This article provides a technical comparison of both methods with an emphasis on its clinical implications. Future dosimetric techniques for Y-90 radioembolization based on emerging technologies are also discussed. PMID- 21643808 TI - Enhanced activity and enantioselectivity of a hyperthermophilic esterase from archaeon Aeropyrum pernix K1 by acetone treatment. AB - To improve the activity and enantioselectivity of hyperthermophilic archaeon Aeropyrum pernix K1 esterase (APE1547) and its mutants, they were purified by acetone-treated method. It was found that the acetone treatment not only caused APE1547 and its mutants to display higher activity and enantioselectivity but also saved more than 90% of time spent in purifying them by Ni-chelating column. In hydrolysis of p-nitrophenyl caprylate, the acetone-treated APE1547 and mutant A containing the following substitutions R11G, L36P, V225A, I551L, and A564T showed 5.7- and 6.9-fold active increase, respectively. In the resolution of 2 octanol acetate, the acetone-treated mutant A had a 9-fold enantioselective increase relative to that purified by Ni-chelating column. In addition, the impact of pH, temperature, and chemical reagents on activity of APE1547 and mutant A was discussed in this paper. PMID- 21643809 TI - Role of circulating tumor cells and disseminated tumor cells in primary breast cancer. AB - Metastasis remains a main cause of death in patients with breast cancer regardless of improvements in treatment. Prospective clinical studies of this minimal residual disease have shown disseminated tumor cells (DTCs) in bone marrow and circulating tumor cells (CTCs) in peripheral blood, neither of which can be detected by conventional imaging, to be prognostic and predictive markers for responsive treatment in patients with metastatic breast cancer. However, the guideline from the American Society of Clinical Oncology does not recommend measuring CTCs for clinical decisions because of a lack of evidence for an established, sound methodology and with proven clinical relevance. The Southwest Oncology Group trial S0500 to validate the clinical relevance of CTCs for treatment decisions in patients with metastatic breast cancer is ongoing. In patients with primary breast cancer, the low detection rate of CTCs has been overcome by recent advances in technology. Although generally DTCs were more detectable than CTCs and the association between presence of DTCs and poor prognosis has been shown, the invasiveness of sample collection of DTCs from bone marrow is generally hard for patients to accept. In this review, we concentrate on the question of whether we need to consider CTCs and DTCs in the management of primary breast cancer on the basis of the evidence of the clinical relevance of CTCs and DTCs. The promising role of the molecular characterization of CTCs, which does have the potential for being a predictor for tumor behavior and development, is suggested as a new targeting strategy. PMID- 21643810 TI - Prevalence of breast arterial calcification by mammography contributes to breast cancer. AB - BACKGROUND: Mammographic breast arterial calcifications (BACs) are regarded as aging-related benign changes in breast cancer screening practices. BACs have recently attracted attention, because several researchers proposed using them as a surrogate marker of arteriosclerosis or osteoporosis. No studies have thus far evaluated an association between BACs and breast cancer, however. METHOD: The percentage of BAC positivity was compared between a group of women aged 40 years or older with breast cancer detected in 2004 through 2009, and a group of women of the same age range in whom breast cancer was not detected by screening mammography in 2009. RESULTS: The BAC-positive rate in the group of 243 breast cancer patients was 9.88% (24 of the 243) and was lower than that in the group of women without breast cancer, 14.34% (506 of 3528 women). The analysis by age revealed that the differences in this parameter between the two groups were significant in women aged 60 years or older. Multivariate analysis including demographic characteristics revealed that when adjusted for age and body weight, BAC positivity was a significant risk factor for breast cancer. CONCLUSION: The study results suggest that BAC positivity and, in turn, arteriosclerosis may have an antinomic relationship with breast cancer. PMID- 21643811 TI - Elastographic evaluation of mucinous carcinoma of the breast. AB - BACKGROUND: Elastography is widely used as a diagnostic tool for the diagnosis of invasive breast cancer. However, no study has yet shown if elastography for diagnosing mucinous carcinoma is as useful as that for diagnosing the usual invasive carcinoma. Mucinous carcinoma is considered as a soft tumor. In this study, we used elastography to evaluate the elasticity of mucinous carcinoma. METHODS: Among 1,015 patients who underwent surgery for primary breast cancer between February 2007 and August 2008 in our facility, the final pathological diagnosis showed only 32 mucinous carcinomas. We evaluated 16 of the 32 mucinous carcinoma patients who underwent preoperative elastography. RESULTS: There were 13 cases of the pure-type and 3 cases of the mixed-type mucinous carcinoma. B mode ultrasound (US) imaging showed mass formation in 16 patients. The elasticity score was 2 in 1 case (8%), 3 in 3 cases (23%), 4 in 7 cases (54%), and 5 in 2 cases (15%). The fat-to-lesion ratio (FLR) was evaluated in 7 cases. The mean value of the FLR was 12 (range 3-30). CONCLUSION: Twelve of the 16 (75%) cases had an elastography score of 4 or 5. Although mucinous carcinoma had an elastography score similar to that of usual invasive carcinoma, elastography may be useful for distinguishing mucinous carcinoma from benign fibroadenoma. PMID- 21643812 TI - Prospective, randomized, single-blind comparison of effects of 6 months' treatment with atorvastatin versus pravastatin on leptin and angiogenic factors in patients with coronary artery disease. AB - Leptin has been reported to exert an atherosclerotic effect by regulating expression of angiogenic factors that have been implicated in the pathogenesis of coronary artery disease (CAD). The purpose of this study was to investigate whether lipid-lowering therapy (LLT) with statins could affect leptin levels and angiogenic factors in patients with CAD. This study included 76 patients with CAD and 15 subjects without CAD (non-CAD). CAD patients were randomized to 6 months of intensive LLT with atorvastatin or moderate LLT with pravastatin. Plasma leptin, angiopoetin-2 (Ang-2), hepatocyte growth factor (HGF) and vascular endothelial growth factor (VEGF) levels were measured prior to statin therapy (baseline) and after 6 months. Baseline levels of leptin, Ang-2, HGF and VEGF were higher in the CAD group than in the non-CAD group (all P < 0.05). Treatment with intensive LLT decreased leptin, Ang-2, HGF and VEGF levels, whereas moderate LLT did not change these levels. This study suggests that LLT with atorvastatin decreases leptin levels and angiogenic factors in patients with CAD, possibly contributing to the beneficial effects of LLT with atorvastatin in CAD. PMID- 21643813 TI - Arrhythmias after transcatheter closure of perimembranous ventricular septal defects with a modified double-disk occluder: early and long-term results. AB - With the development of interventional techniques and devices, transcatheter closure of perimembranous ventricular septal defect has been widely performed. However, there has been a lack of long-term follow-up results about postoperative ECG changes of PmVSD patients. We report our experience of early and late arrhythmias after transcatheter closure of PmVSD with a modified double-disk occluder (MDVO). We performed a retrospective review of 79 patients (47 males, 32 females) between September 2002 and May 2007 who underwent transcatheter closure of perimembranous ventricular septal defect. Symmetric and asymmetric PmVSD occluders were used. The diameter of the evaluated defects ranged from 3 to 12 mm, as measured by TTE and 3 to 15 mm by left ventriculography. Most cases of PmVSD were treated successfully with a single procedure, resulting in a successful closure rate of 97% (77/79 patients). There was no death in any of the patients. After the operation, 79 patients were followed-up for a range of 10-76 months (35.3 +/- 17.4 months). In this series, 11 cases of incomplete right bundle branch block and five cases of complete right bundle branch block occurred during the early period after operation. During long-term follow-up, these issues declined in prevalence to five and four cases, respectively. Moreover, reversible third-degree AVB occurred during closure or after the procedure, and two of the three patients with reversible AVB received a temporary heart pacemaker implantation. These patients recovered 1 h, 6 days, and 9 days later, respectively. During 10-76 months of follow-up, no complications occurred in any of the patients, including residual shunt, severe aortic valve, or tricuspid valve regurgitation. Device closure of perimembranous ventricular septal defects with a modified double-disk occluder (MDVO) resulted in excellent closure rates and acceptably low arrhythmia rates. PMID- 21643814 TI - Do inexperienced bumblebee foragers use scent marks as social information? AB - Bumblebees (Bombus spp.) foraging in the field typically reject flowers where they detect the olfactory footprints of previous visitors and hence avoid recently emptied inflorescences. A growing number of studies have begun to illustrate that associative learning shapes the development of this process, in both bumblebees and other bee species. This raises the question of what the default response to such marks is, but little is known about how inexperienced foragers use social information. Here, we offered flower-naive bees a choice between scent-marked flowers and unmarked alternatives and found that individuals neither avoided nor preferred marked flowers. Our findings provide no support for 'hard-wired' responses to scent marks in bumblebees and highlight the importance of associative learning in shaping social information use to match local circumstances. PMID- 21643815 TI - Variation in GUS activity in vegetatively propagated Hevea brasiliensis transgenic plants. AB - Hevea brasiliensis transgenic plants are regenerated from transgenic callus lines by somatic embryogenesis. Somatic embryogenesis is not yet available for commercial propagation of Hevea clones, which requires conventional grafting of buds on rootstock seedlings (budding). The stability of transgene expression in budded plants is therefore necessary for further development of genetic engineering in rubber trees. Transgene expression was assessed by fluorimetric beta-glucuronidase (GUS) activity in fully developed leaves of in vitro plants from transgenic lines and their sub-lines obtained by budding. A large variation in GUS activity was found in self-rooted in vitro plants of five transgenic lines, and the absence of activity in one line suggested transgene silencing. Beyond confirming transmissibility of the reporter gene by budding and long-term expression, a quantification of GUS activity revealed that greater variability existed in budded plants compared to self-rooted mother in vitro plants for three transgenic lines. Although somatic embryogenesis provided more stable GUS activity, budding remained an efficient way of propagating transgenic plants but transgene expression in budded plants should be verified for functional analysis and further development. PMID- 21643816 TI - Exposure to a commercial glyphosate formulation (Roundup(r)) alters normal gill and liver histology and affects male sexual activity of Jenynsia multidentata (Anablepidae, Cyprinodontiformes). AB - Roundup is the most popular commercial glyphosate formulation applied in the cultivation of genetically modified glyphosate-resistant crops. The aim of this study was to evaluate the histological lesions of the neotropical native fish, Jenynsia multidentata, in response to acute and subchronic exposure to Roundup and to determine if subchronic exposure to the herbicide causes changes in male sexual activity of individuals exposed to a sublethal concentration (0.5 mg/l) for 7 and 28 days. The estimated 96-h LC50 was 19.02 mg/l for both male and female fish. Gill and liver histological lesions were evaluated through histopathological indices allowing quantification of the histological damages in fish exposed to different concentrations of the herbicide. Roundup induced different histological alterations in a concentration-dependent manner. In subchronic-exposure tests, Roundup also altered normal histology of the studied organs and caused a significant decrease in the number of copulations and mating success in male fish exposed to the herbicide. It is expected that in natural environments contaminated with Roundup, both general health condition and reproductive success of J. multidenatata could be seriously affected. PMID- 21643817 TI - Genetic structure and diversity of wild sorghum populations (Sorghum spp.) from different eco-geographical regions of Kenya. AB - Wild sorghums are extremely diverse phenotypically, genetically and geographically. However, there is an apparent lack of knowledge on the genetic structure and diversity of wild sorghum populations within and between various eco-geographical regions. This is a major obstacle to both their effective conservation and potential use in breeding programs. The objective of this study was to assess the genetic diversity and structure of wild sorghum populations across a range of eco-geographical conditions in Kenya. Sixty-two wild sorghum populations collected from the 4 main sorghum growing regions in Kenya were genotyped using 18 simple sequence repeat markers. The study showed that wild sorghum is highly variable with the Coast region displaying the highest diversity. Analysis of molecular variance showed a significant variance component within and among wild sorghum populations within regions. The genetic structure of wild sorghum populations indicated that gene flow is not restricted to populations within the same geographic region. A weak regional differentiation was found among populations, reflecting human intervention in shaping wild sorghum genetic structure through seed-mediated gene flow. The sympatric occurrence of wild and cultivated sorghums coupled with extensive seed-mediated gene flow, suggests a potential crop-to-wild gene flow and vice versa across the regions. Wild sorghum displayed a mixed mating system. The wide range of estimated outcrossing rates indicate that some environmental conditions may exist where self-fertilisation is favoured while others cross-pollination is more advantageous. PMID- 21643818 TI - Endoscopic stenting for post-transplant biliary stricture: usefulness of a novel removable covered metal stent. AB - BACKGROUND: Endoscopic management of biliary anastomotic stricture (AS) following liver transplantation (LT) remains challenging. There are no dedicated self expandable metal stents (SEMS) for this setting. METHODS: A short fully covered SEMS (FCSEMS) with a retrieval suture was designed. Between July 2008 and June 2010, 13 patients with post-LT AS had this FCSEMS placed endoscopically, keeping the whole stent inside the bile duct across the AS with the retriever out of the papilla. The stents were removed by forceps under endoscopy according to a schedule. Technical success, complications, AS resolution and the outcome for the patients were observed. RESULTS: Placement of the FCSEMS was successful on the first attempt in all patients. One patient with complicated infection did not respond to the stenting therapy and underwent stent retrieval ahead of schedule. Others kept well during stenting for a mean (SD) duration of 5.4 (1.7) months (range 2-8) without stent migration. All stents were removed successfully without great difficulty. AS resolution was obtained in all 12 patients, who were closely followed up for a mean (SD) time of 12.1 (8.0) months (range 1-26.5) after stent removal. Stricture recurrence occurred in one, who underwent a successful re intervention with a second FCSEMS. Others remain free from symptoms and have normal liver function up to now. CONCLUSIONS: Endoscopic treatment of post-LT AS using a removable FCSEMS is technically feasible, safe, and effective. This dedicated method may play an increasing role in the future management of benign biliary strictures. PMID- 21643819 TI - Evolution of PVA gels prepared without crosslinking agents as a cell adhesive surface. AB - Physical parameters (such as crosslinking density, crystallinity and mechanical properties) have been found to largely affect cellular behavior on polymer scaffolds. This study demonstrated that transparent pure Poly (vinyl alcohol) hydrogels prepared via a freeze-thaw method can be made to support cell adhesion by controlling physical parameters such as concentration and the number of freeze thaw cycles. For a given number of freeze-thaw cycles, (specifically 45), polymer concentration dependent structural and mechanical properties (such as tensile strength and stiffness) were correlated with cell adhesion. The maximum cell attachment occurred on the hydrogels with the greatest mechanical properties, crystallinity and crosslinking density. The hydrogel surfaces were more favorable to human dermal fibroblasts than human lens epithelial cells and retained their transparency as well as dimensional stability with only a small degree of swelling. Fibroblast laden hydrogels showed extensive alkaline phosphatase activity which confirmed their healthy proliferation and function. In this manner, this study suggests that transparent Poly (vinyl alcohol) hydrogels prepared by the freeze thaw method described here should be further studied for numerous tissue engineering applications. PMID- 21643820 TI - Carcinoma cuniculatum arising in the tongue. AB - Carcinoma cuniculatum (CC) is a rare, distinct clinico-pathological variant of squamous cell carcinoma (SCC) that is defined histologically by the characteristic infiltrative pattern of a deep, broad, and complex proliferation of stratified squamous epithelium with keratin cores and keratin-filled crypts. Herein, we present a case report of CC of the oral tongue and discuss its diagnosis, management, and outcome, as well as briefly review the world literature. To our knowledge, this is the first documented case of CC of the tongue to be reported in the English literature. We draw attention to its clinico pathological features and highlight that awareness of this entity as a distinct variant of SCC facilitates its correct management. PMID- 21643821 TI - Thromboprophylaxis use in medical and surgical inpatients and the impact of an electronic risk assessment tool as part of a multi-factorial intervention. A report on behalf of the elVis study investigators. AB - Venous thromboembolism (VTE) is a major source of morbidity and mortality for both surgical and medical hospitalised patients. Despite the availability of guidelines, thromboprophylaxis continues to be underutilised. This study aims to assess the effectiveness of an electronic VTE risk assessment tool (elVis) on VTE prophylaxis in hospitalised patients. A national, multicentre, prospective clinical audit collected information on VTE prophylaxis and risk factors for VTE in 2,400 hospitalised patients (comprising of equal numbers of medical, surgical and orthopaedic patients). After auditing the standard care use of VTE prophylaxis in 1,200 consecutive patients (audit 1, A1), the elVis system was installed and a second audit (A2) of VTE prophylaxis was performed in a further 1,200 patients. The use of the electronic VTE risk assessment tool was low with 20.5% of patients assessed with elVis. The intervention, elVis plus accompanying education, improved the use VTE prophylaxis to guidelines by 5.0% amongst all patients and by 10.7% amongst high risk patients (adjusted odds ratio (AOR) 1.27 and 1.65 respectively). The use of elVis in A2 varied between hospitals and specialties and this resulted in marked heterogeneity. Despite this heterogeneity, patients assessed with elVis had 1.44 times higher AOR of being treated to guidelines compared to those who were not (P < 0.05). The use of elVis accompanied by staff education improved VTE prophylaxis, especially amongst high risk patients. To optimise the effectiveness and support enduring practice change electronic systems, such as elVis, need to be completely integrated within the treatment pathway. PMID- 21643822 TI - Assessing readiness for establishing a farmers' market at a community health center. AB - Farmers' markets are community health promotion interventions that increase access to fresh fruits and vegetables. As farmers' markets continue to develop, it is important to strategically locate them in settings that are accessible to populations disparately affected by health disparities. One potential setting is a community health center. The goal of this analysis is to extend existing research on community readiness to identify indicators of preparedness among community health centers for establishing onsite farmers' markets. The sampling frame for the readiness assessment included all community health centers in South Carolina (N = 20) representing 163 practice sites. Data collection included two brief online surveys, in-depth key informant interviews, and secondary analysis of contextual data. Five themes related to readiness for establishing a farmers market at a community health center were identified: capacity, social capital, awareness of health problems and solutions, logistical factors, and sustainability. Findings from this study provide guidance to researchers and community health center staff as they explore the development of environmental interventions focused on reducing diet-related health conditions by improving access to healthy foods. PMID- 21643823 TI - Prevalence of HIV risk behaviors between binge drinkers and non-binge drinkers aged 18- to 64-years in US, 2008. AB - Using data from the 2008 Behavioral Risk Factor Surveillance System on 281,303 adults aged 18-64 years in the United States, we examined the relationship between HIV risk behaviors and binge drinking of alcoholic beverages and the frequency of binge drinking among a subgroup of 41,073 respondents who were acknowledged binge drinkers (bingers), based on reported drinking behavior in the year preceding survey. Our findings show that the weighted prevalence of HIV risk behaviors (including injection drug use, exchange of sex for money/drugs, and anal sex without a condom) among binge drinkers [corrected] [7.0%, 95% confidence interval (95% CI): 6.4-7.6%] is twice that among nonbingers (2.9%, 95% CI: 2.7 3.0%). The highest prevalence of HIV risk behaviors is among the bingers aged 18 20 years (14%, 95% CI: 11.2-18.2%). After adjusting for covariates, bingers are 1.77 (95% CI: 1.58-2.00) times more likely than nonbingers to report HIV risk behaviors. Risk increases in bingers with the number of episodes. Compared with bingers reporting 1-2 binge episodes in the month proceeding survey, the adjusted odds of reporting HIV risk behaviors among bingers are 1.27 (1.08-1.49), 1.68 (1.35-2.10), 1.67 (1.08-2.57), and 1.70 (1.34-2.16), respectively for bingers with 3-4, 5-6, 7-8, and >=9 episodes in the same period. Our results suggest that HIV risk behaviors are strongly linked with binge drinking and its frequency. Effective measures to prevent binge drinking are essential to HIV prevention, especially among youth aged 18-20 years. PMID- 21643824 TI - Tap or bottled water: drinking preferences among urban minority children and adolescents. AB - The last decade has seen an increasing trend in consumer preference of bottled water over tap water. Little is known what type of water children and adolescents prefer for drinking and what their parents think of their community tap water. The study objective was to assess drinking water preferences, perceptions of the qualities of tap water and bottled water, and fluoride knowledge in an urban pediatric population. We conducted an anonymous survey of a convenience sample of caretakers of children and adolescents at an urban clinic regarding their preferences for tap or bottled water, their perceptions of the quality of tap and bottled water and their knowledge of fluoride. Of the 208 participants (79% African American, 9% Latino), 59% drank tap water, 80% bottled water. Only 17% drank tap water exclusively, 38% drank bottled water exclusively, 42% drank both. We found no significant differences in water preferences across age groups, from infancy to adulthood, or among ethnic groups. Ratings for taste, clarity, purity and safety were significantly higher for bottled water than tap water (P < 0.001). Only 24% were aware of fluoride in drinking water. We conclude bottled water was preferred over tap water in an urban minority pediatric population. Perceptions of the qualities of water seemed to drive drinking preferences. Public health strategies are needed to increase public awareness of the impact of bottled water consumption on oral health, household budgets and the environment. PMID- 21643825 TI - Anterior single screw fixation of odontoid fracture with intraoperative Iso-C 3 dimensional imaging. AB - PURPOSE: The purpose of this study was to assess the value of isocentric C-arm three-dimensional (Iso-C 3D) fluoroscopy for the insertion of an anterior odontoid screw. The results of the Iso-C 3D group were compared with that of an historic control group using conventional fluoroscopy. METHODS: Twenty-nine patients diagnosed with type II or rostral-type III odontoid fractures were treated with a single anterior screw fixation in this study. The Iso-C 3D group included 13 patients and the other 16 patients were in the historic control group. All operations were performed by a single surgeon using standard procedure and manner. The clinical and radiographic results were recorded and compared between the two groups. RESULTS: The fluoroscopy time in the Iso-C 3D group was 42.9 s as compared to 68.1 s in the control group (P < 0.01). The mean operative time was 91.5 min in the Iso-C 3D group when compared with 81.6 min in the control group (P = 0.20). The rate of bony fusion was 96.6% (28/29), the failure rate of reduction or fixation was 13.8% (7.7% in Iso-C 3D group; 18.8% in control group). The Smiley-Webster scale showed that 90% of patients achieved good or better outcomes CONCLUSIONS: In conclusion, this technique can be safely extended to the treatment of technically difficult to treat spinal injuries and at the same time reduce total radiation exposure time both for the patient and the surgeon. PMID- 21643826 TI - Anterior migration of spinal cord after cervical corpectomy. AB - Posterior migration of spinal cord is a common complication of posterior cervical decompression, whereas anterior migration of spinal cord after anterior cervical decompression has not been reported previously. This report presents a case of anterior migration of spinal cord after cervical corpectomy. A 65-year-old male underwent a cervical corpectomy of the C4 and C5 for cervical spondylotic myelopathy. The postoperative MRI showed that spinal cord was decompressed. However, at 12 months postoperatively, the patient complained that improved gait aggravated again, and the MRI showed an anterior migration of the spinal cord at the level of the C4 and C5, and a compression of spinal cord at the level of caudal endplate of C3. Secondary surgery, laminaplasty of C3 and C4 was conducted. The postoperative MRI showed that the spinal canal increased at the C3 and C4 levels, and the spinal cord went through smoothly without sharp turning. PMID- 21643827 TI - Molecular evolution of mollusc shell proteins: insights from proteomic analysis of the edible mussel Mytilus. AB - Shell matrix proteins (SMPs) that are embedded within calcified layers of mollusc shells are believed to play an essential role in controlling the biomineral synthesis and in increasing its mechanical properties. Among the wide diversity of mollusc shell textures, nacro-prismatic shells represent a tremendous opportunity for the investigation of the SMP evolution. Indeed, nacro-prismatic texture appears early in Cambrian molluscs and is still present in the shell of some bivalves, gastropods, cephalopods and very likely also, of some monoplacophorans. One key question is to know whether these shells are constructed from similar matrix protein assemblages, i.e. whether they share a common origin. Most of the molecular data published so far are restricted to two genera, the bivalve Pinctada and the gastropod Haliotis. The shell protein content of these two genera are clearly different, suggesting independent origins or considerable genetic drift from a common ancestor. In order to describe putatively conserved mollusc shell proteins, here we have investigated the SMP set of a new bivalve model belonging to another genera, the edible mussel Mytilus, using an up-to-date proteomic approach based on the interrogation of more than 70,000 EST sequences, recently available from NCBI public databases. We describe nine novel SMPs, among which three are completely novel, four are homologues of Pinctada SMPs and two are very likely homologues of Haliotis SMPs. This latter result constitutes the first report of conserved SMPs between bivalves and gastropods. More generally, our data suggest that mollusc SMP set may follow a mosaic pattern within the different mollusc models (Mytilus, Pinctada, Haliotis). We discuss the function of such proteins in calcifying matrices, the molecular evolution of SMP genes and the origin of mollusc nacro prismatic SMPs. PMID- 21643828 TI - Evolution of vertebrate immunity: sequence and functional analysis of the SEFIR domain family member Act1. AB - SEF/IL-17R/CIKS/ACT1 homology (SEFIR) domain containing proteins, which include the IL-17 receptors and an adaptor protein Act1, have essential roles in vertebrate immunity. However, the molecular mechanisms of Act1 function remain largely unexplored. In this article, we employed an evolutionary analysis to discover novel structural and functional properties of Act1. Firstly, we have found that the previously identified helix-loop-helix and Ufd2-box domains in human Act1 have relatively recent evolutionary origins in higher vertebrates. Zebrafish Act1, which lacks these domains, is unable to induce JNK phosphorylation and activate cytokine expression when expressed in human cells. Secondly, we have established that Act1-like proteins contain DEATH-domains in basal animals, such as Hydra and primitive chordates, but lack this domain in vertebrates. Finally, we have shown that Act1-TRAF6 interactions are conserved throughout vertebrate evolution: Act1 derived from zebrafish can bind to TRAF6 and activate NF-kappaB in human cells. Moreover, we have identified a novel highly conserved motif at the amino-terminus of Act1, which is critical for binding to TRAF6 and activating NF-kappaB-dependent gene expression. We propose a model of evolutionary changes in Act1-mediated signalling, which contributes to a better understanding of evolution of the vertebrate immunity. PMID- 21643829 TI - Absence of positive selection on centromeric histones in Tetrahymena suggests unsuppressed centromere: drive in lineages lacking male meiosis. AB - Centromere-drive is a process where centromeres compete for transmission through asymmetric "female" meiosis for inclusion into the oocyte. In symmetric "male" meiosis, all meiotic products form viable germ cells. Therefore, the primary incentive for centromere-drive, a potential transmission bias, is believed to be missing from male meiosis. In this article, we consider whether male meiosis also bears the primary cost of centromere-drive. Because different taxa carry out different combinations of meiotic programs (symmetric + asymmetric, symmetric only, asymmetric only), it is possible to consider the evolutionary consequences of centromere-drive in the context of these differing systems. Groups with both types of meiosis have large, rapidly evolving centromeric regions, and their centromeric histones (CenH3s) have been shown to evolve under positive selection, suggesting roles as suppressors of centromere-drive. In contrast, taxa with only symmetric male meiosis have shown no evidence of positive selection in their centromeric histones. In this article, we present the first evolutionary analysis of centromeric histones in ciliated protozoans, a group that only undergoes asymmetric "female" meiosis. We find no evidence of positive selection acting on CNA1, the CenH3 of Tetrahymena species. Cytological observations of a panel of Tetrahymena species are consistent with dynamic karyotype evolution in this lineage. Our findings suggest that defects in male meiosis, and not mitosis or female meiosis, are the primary selective force behind centromere-drive suppression. Our study raises the possibility that taxa like ciliates, with only female meiosis, may therefore undergo unsuppressed centromere drive. PMID- 21643830 TI - Dissipation and residue of thiacloprid in cabbage and soil. AB - Residue dynamics of thiacloprid in cabbage and soil was studied in this paper utilizing liquid chromatography with tandem mass spectrometry (LC-MS/MS). The field trial was conducted in two sites: Beijing, China and Hubei, China. Thiacloprid dissipated rapidly with the half-life 1.3-1.6 days in cabbage and 2.1 3.1 days in soil. In the terminal residue experiment, no higher residue than 0.06 mg/kg in cabbage and 0.16 mg/kg in soil was detected, which was far below either EU MRL (0.2 mg/kg) or Japan MRL (1 mg/kg). PMID- 21643831 TI - Toxicity of phenanthrene in freshwater sediments to the rooted submersed macrophyte, Vallisneria spiralis. AB - A study was conducted to determine the response of the rooted submersed macrophyte, Vallisneria spiralis to phenanthrene in freshwater sediments with initial phenanthrene concentrations from 0 to 80 mg kg(-1) dry sediment. The sensitivity of various morphological endpoints was evaluated after 90 days of exposure. The most sensitive toxicity test endpoints were those that reflected root growth. Toxicological sensitivity of the endpoints changed with the effect level selected. The toxicity threshold from a plot of the EC(10) values was 1-2 orders of magnitude lower than those calculated for the threshold from plots of the EC(25) or EC(50) values. In addition, stimulatory responses (hormesis) on root growth were observed at subtoxic concentrations of phenanthrene, and a hormetic model should thus be incorporated for ecological risk assessment. PMID- 21643832 TI - Isolation, selection and biodegradation profile of phenol degrading bacteria from oil contaminated soil. AB - In the present study, an aerobic bacterial strains OCS-A and OCS- B were isolated from an oil contaminated soil. The strains were identified to be Citrobacter freundi and Proteus mirabilis according to morphological, physiological and biochemical characteristics. The strains were able to degrade about 90% of 100 mg/L phenol within 80 h as sole carbon and energy source. The lag phase increased with increase in phenol concentration. Determination of metabolic intermediate 2 HMS, was done which indicate meta-cleavage pathway of phenol metabolism. Hence these isolates can be effectively used for bioremediation of phenol contaminated sites. PMID- 21643833 TI - Unequal chromosome division and inter-genomic translocation occurred in somatic cells of wheat-rye allopolyploid. AB - Newly synthesized wheat-rye allopolyploids were investigated by genomic in situ hybridization, over the first, second, third and fourth allopolyploid generations. Inter and intra chromosome connections were observed in 12 root-tip cells of CA4.4.7 (S(2) generation), and translocations between wheat and rye chromosomes were also detected in five root-tip cells. In root-tip cells of CA4.4.7.5 and CA4.4.7.2.2 (S(3) and S(4) generation), the chromosome connections occurred again, a dissociative small rye segment was detected in seven cells of CA4.4.7.5. In plants MSV6.1 and MSV6.5 (S(1) generation), almost half of the root tip cells contained 13 rye chromosomes and the rest held 12 rye chromosomes, and all the cells of the two plants contained 42 wheat chromosomes. Five pairing configurations of rye chromosomes, including 5 II + 3 I, 6 II + 1 I, 6 II, 5 II + 2 I and 4 II + 4 I, were observed in pollen mother cells of the two plants. The two plants' progeny, including S(2), S(3), and S(4) generation plants, contained 42 wheat chromosomes and 12 rye chromosomes. Therefore, the inter chromosome translocation and unequal chromosome division could occur in somatic cells of wide hybrids. The unequal chromosome division in somatic cell could induce chromosome elimination at the early stages of allopolyploidization. PMID- 21643834 TI - Age of achievement of gross motor milestones in infancy and adiposity at age 3 years. AB - Early life physical activity may help prevent obesity but is difficult to measure. The purpose of this study was to examine associations of age of achievement of gross motor milestones in infancy with adiposity at age 3 years. Seven forty one mother/infant dyads participated in a longitudinal study in Massachusetts. Exposures were age of attainment of 4 gross motor milestones rolling over, sitting up, crawling, and walking. Outcomes were 3-year sum of subscapular and triceps skinfold thickness (SS + TR) for overall adiposity, their ratio (SS:TR) for central adiposity, and body mass index (BMI) z-score. We used linear regression models adjusted for confounders to examine motor milestone achievement and later adiposity. Rolling over (0.04, 95% CI: 0.008, 0.07) and sitting up (0.02, 95% CI: 0.001, 0.05) at >= 6 months were associated with increased SS:TR compared with attainment before 6 months. Walking at >= 15 months was associated with 0.98 mm higher SS + TR (95% CI: 0.05, 1.91) compared with walking before 12 months. Age at crawling was not associated with the outcomes. None of the milestones were associated with BMI z-score. Age of motor milestone achievement was only a modest predictor of adiposity. Later rolling over and sitting up were associated with greater central adiposity, and later age at walking was associated with greater overall adiposity at age 3 years. Although we controlled for birth weight and 6-month weight-for-length in our models, more detailed assessment of early adiposity prior to achievement of motor milestones is needed to help determine causality. PMID- 21643835 TI - 1H, 15N and 13C assignments of an intramolecular LMO4-LIM1/CtIP complex. AB - LMO4 is a broadly expressed LIM-only protein that is involved in neural tube development and implicated in breast cancer. Here we report backbone and side chain NMR assignments for an engineered intramolecular complex of the N-terminal LIM domain from LMO4 tethered to residues 641-685 of C-terminal binding protein interacting protein (CtIP/RBBP8). PMID- 21643836 TI - Survivors of breast cancer: patient perspectives on survivorship care planning. AB - INTRODUCTION: Survivors of breast cancer (SBC) constitute the largest population of cancer survivors. Needs for survivorship care may vary according to life stage and urban/rural place of residence. This study was conducted to better understand patient preferences for survivorship care plans (SCP). METHODS: Patients were identified through cancer agency electronic records and invited to participate by mail. Sessions were stratified by age and rural/urban place of residence. Participants were asked about preferences for content and format of SCP. Focus groups were conducted using a semi-structured format with interviews being taped, transcribed, cross-checked for accuracy, and analyzed independently, using constant comparative methods. RESULTS: Preferred SCP key elements included treatment summary, information on nutrition/exercise, expected side effects, signs and symptoms of recurrence, recommended follow-up schedule, information sent to primary care physician, and updates on changes. SBC emphasized preference for individualized content depending upon physical and psychosocial effects. No difference was observed between preferred SCP content among patients residing in urban/rural areas. Rural participants preferred electronic formats for ongoing information bulletins and communication with health care providers. DISCUSSION/CONCLUSIONS: SBC from all age groups identify common preferences for key SCP elements with individualized content reflecting the wide variation observed among physical and psychosocial effects of breast cancer. Patterns of key psychological, social, and physical effects observed at different life stages may help SCP customization. IMPLICATIONS FOR CANCER SURVIVORS: Results provide direction for designing key content and format of SCP and also provide information about elements of care planning that should be customized to individual patient needs. PMID- 21643837 TI - What's wrong with the tap? Examining perceptions of tap water and bottled water at Purdue University. AB - The environmental impacts of bottled water prompted us to explore drinking water choices at Purdue University, located in West Lafayette, IN. A random sample of 2,045 Purdue University students, staff, and faculty was invited to participate in an online survey. The survey assessed current behaviors as well as perceived barriers and benefits to drinking tap water versus bottled water. 677 surveys were completed for a response rate of 33.1%. We then conducted qualitative interviews with a purposive sample of university undergraduates (n = 21) to obtain contextual insights into the survey results and the beliefs of individuals with a variety of drinking water preferences. This study revealed that women drink disproportionately more bottled water then men while undergraduate students drink more than graduate students, staff and faculty. The study also uncovered a widespread belief that recycling eliminates the environmental impacts of bottled water. Important barriers to drinking tap water at Purdue include: perceived risks from tap water and the perceived safety of bottled water, preferring the taste of bottled water, and the convenience of drinking bottled water. The qualitative interviews revealed that drinking water choices can be influenced by several factors-especially whether individuals trust tap water to be clean-but involve varying levels of complexity. The implications of these results for social marketing strategies to promote tap water are discussed. PMID- 21643838 TI - Dietary conjugated linoleic acid induces lipolysis in adipose tissue of coconut oil-fed mice but not soy oil-fed mice. AB - Mice fed diets containing conjugated linoleic acid (CLA) are leaner than mice not fed CLA. This anti-obesity effect is amplified in mice fed coconut oil-containing or fat free diets, compared to soy oil diets. The present objective was to determine if CLA alters lipolysis in mice fed different base oils. Mice were fed diets containing soy oil (SO), coconut oil (CO), or fat free (FF) for 6 weeks, followed by 10 or 12 days of CLA or no CLA supplementation. Body fat, tissue weights, and ex vivo lipolysis were determined. Relative protein abundance and activation of perilipin, hormone sensitive lipase (HSL), adipose triglyceride lipase (ATGL), and adipose differentiation related protein (ADRP) were determined by western blotting. CLA feeding caused mice to have less (P < 0.05) body fat than non-CLA fed mice. This was enhanced in CO and FF-fed mice (CLA * oil source, P < 0.05). There was also a CLA * oil source interaction on lipolysis as CO + CLA and FF + CLA-fed mice had increased (P < 0.05) rates of lipolysis but SO + CLA fed mice did not. However, after 12 days of CLA consumption, activated perilipin was increased (P < 0.05) only in SO + CLA-fed mice and total HSL and ATGL were decreased (P < 0.05) in CO + CLA-fed mice. Therefore, the enhanced CLA-induced body fat loss in CO and FF-fed mice appears to involve increased lipolysis but this effect may be decreasing by 12 days of CLA consumption. PMID- 21643839 TI - Comment to the paper: Palliative functional hemispherectomy for treatment of refractory status epilepticus associated with Alper's disease. PMID- 21643840 TI - Ethynyldeoxyuridine (EdU) suppresses in vitro population expansion and in vivo tumor progression of human glioblastoma cells. AB - Thymidine analogs (TAs) are synthetic nucleosides that incorporate into newly synthesized DNA. Halogenated pyrimidines (HPs), such as bromodeoxyuridine (BrdU), are a class of TAs that can be detected with antibodies and are commonly used for birthdating individual cells and for assessing the proliferative index of cell populations. It is well established that HPs can act as radiosensitizers when incorporated into DNA chains, but they are generally believed not to impair normal cell function in the absence of secondary stressors. However, we and others have shown that HP incorporation leads to a sustained suppression of cell cycle progression in mammalian cells, resulting in cellular senescence in somatic cells. In addition, we have shown that HP incorporation results in delayed tumor progression in a syngeneic rat model of glioma. Here we examine ethynyldeoxyuridine (EdU), a newly developed and alkylated TA, for its anti cancer activity, both in vitro and in vivo. We show that EdU, like HPs, leads to a severe reduction in the proliferation rate of normal and transformed cells in vitro. Unlike HPs, however, EdU incorporation also causes DNA damage resulting in the death of a substantial subset of treated cells. When administered over an extended time as a monotherapy to mice bearing subcutaneous xenografts of human glioblastoma multiforme tumors, EdU significantly reduces tumor volume and increases survival without apparent significant toxicity. These results, combined with the fact that EdU readily crosses the blood-brain barrier, support the continued investigation of EdU as a potential therapy for malignant brain tumors. PMID- 21643841 TI - Cerebellar glioblastoma: a retrospective review of 21 patients at a single institution. AB - Primary cerebellar glioblastoma (CGB) comprises only 0.4-3.4% of all intracranial glioblastoma. The impact of surgical resection on survival and the efficacy of adjuvant therapies are uncertain as CGB is underrepresented in most studies. To elucidate prognostic factors we performed a single-institutional review of the largest series to date of CGB. The University of Texas MD Anderson Cancer Center database was reviewed from 1990 to 2010. Twenty-one consecutive patients met criteria for inclusion. The Kaplan-Meier product limit method was used to estimate overall survival (OS) and progression-free survival (PFS); groups were compared using the log-rank statistic. The multivariate Cox proportional hazards models were fitted to examine the association of resection with OS and PFS adjusted for other clinical variables. The median age was 39.9 years, and Karnofsky performance status (KPS) was >= 80 in 61.5% of patients. The mean extent of resection for contrast enhancement (EOR-CE) was 93.8% (SD = 10.4%; median = 100%), and the median follow-up was 18.4 months (range 1.5-116.1 months). There was no significant association of EOR with OS or PFS. On univariate analysis the presence of leptomeningeal disease (LMD) was associated with a worse OS (6.1 vs. 24.1 months; P = 0.0001) and PFS (3.3 vs. 9 months; P = 0.019). Patients who had adjuvant chemotherapy (CT) had extended PFS (10.1 vs. 2.8 months; P < 0.0001). Adjustment for the presence of leptomeningeal disease (LMD) tended toward an increased risk of progression (HR = 3.46; 95% confidence interval [CI], 0.83-14.5; P = 0.09) and was associated with a significantly increased risk of death (HR = 15.2; 95% CI, 1.3-180; P = 0.03). Having received adjuvant chemotherapy was associated with a decreased risk of progression (HR = 0.02; 95% CI, 0-0.26; P = 0.003). The presence of LMD is a critical factor in the clinical behavior of CGB resulting in markedly decreased OS and PFS. Adjuvant CT resulted in increased PFS but did not significantly affect OS. This was due to a lack of a sizable cohort who did not receive chemotherapy. Furthermore, three of the CT-naive patients received CT at first progression. In the context of the high EOR in this study, an OS of 18.4 months was achieved. PMID- 21643842 TI - IDH1 and IDH2 mutations, immunohistochemistry and associations in a series of brain tumors. AB - A total of 343 brain tumors were studied for IDH1 and IDH2 mutations by direct sequencing and for protein expression by immunohistochemistry with mIDH1(R132H) antibody. Of these, 287 were gliomas (17 pilocytic astrocytomas, 13 grade II and 5 grade III astrocytomas, 167 primary (pGBMs) and 19 secondary (sGBMs) glioblastomas, 36 grade II and 26 grade III oligodendrogliomas and 4 grade II-III oligoastrocytomas). In gliomas, IDH1 mutations at codon R132 were identified in 22.3%, of which 93.7% were c.395G>A (p.R132H). Mutations were more frequent in oligodendrogliomas (53.2%) than in astrocytic tumors (22.8%) and in sGBMs (84.2%) upon pGBMs (1.8%). There was a statistically significant correlation between mIDH1(R132H) antibody immunostaining and the relevant mutation c.395G>A (p.R132H) (P = 0.0001). No mutations were identified in non-glial tumors which were also negative to immunohistochemistry, with the exception of one PNET. A c.515G>T (p.R172M) mutation of the IDH2 gene was only identified in a grade II oligodendroglioma patient which was wild-type for IDH1. A direct correlation with MGMT promoter hypermethylation status and an inverse correlation with EGFR amplification was found, whereas the relationships with 1p/19q co-deletion and TP53 mutations only showed a trend toward correlation. In all gliomas, a positive correlation was found between IDH1 mutations and a young age (P = 0.0001). In contrast, a correlation with overall survival could only be obtained in low-grade gliomas. Immunohistochemistry appeared to be useful in differential diagnoses, especially toward non-tumor pathologic nervous tissue, and in recognizing infiltrating glioma cells. The mIDH1(R132H) antibody positivity was complementary with Cyclin D1 expression. PMID- 21643843 TI - Photodynamic antifungal therapy against chromoblastomycosis. AB - Photodynamic therapy (PDT) is a minimally invasive approach, in which a photosensitizer compound is activated by exposure to light. The activation of the sensitizer drug results in several chemical reactions, such as the production of reactive oxygen species and other reactive molecules, which presence in the biological site leads to the damage of target cells. Although PDT has been primarily developed to combat cancerous lesions, this therapy can be employed for the treatment of several conditions, including infectious diseases. A wide range of microorganisms, including Gram-positive and Gram-negative bacteria, viruses, protozoa, and fungi, have demonstrated susceptibility to antimicrobial PDT. This treatment might consist in an alternative for the management of fungal infections. Antifungal photodynamic therapy has been successfully employed against Candida species, dermatophytes, and Aspergillus niger. Chromoblastomycosis is an infection that involves skin and subcutaneous tissues caused by the traumatic inoculation of dematiaceous fungi species, being that the most prevalent are Fonsecaea pedrosoi and Claphialophora carrionii. In the present work, the clinical applications of PDT for the treatment of chromoblastomycosis are evaluated. We have employed methylene blue as photosensitizer and a LED (Light Emitting Diode) device as light source. The results of this treatment are positive, denoting the efficacy of PDT against chromoblastomycosis. Considering that great part of the published works are focused on in vitro trials, these clinical tests can be considered a relevant source of information about antifungal PDT, since its results have demonstrated to be promising. The perspectives of this kind of treatment are analyzed in agreement with the recent literature involving antifungal PDT. PMID- 21643844 TI - Identification of a lysine residue important for the catalytic activity of yeast farnesyl diphosphate synthase. AB - The Saccharomyces cerevisiae ERG20 gene (encoding farnesyl diphosphate synthase) has been subjected to a set of mutations at the catalytic site, at position K254 to determine the in vivo impact. The mutated strains have been shown to exhibit various growth rates, sterol profiles and monoterpenol producing capacities. The results obtained suggest that K at position 254 helps to stabilize one of the three Mg(2+) forming a bridge between the enzyme and DMAPP, and demonstrate that destabilizing two of the three Mg(2+) ions, by introducing a double mutation at positions K197 and K254, results in a loss of FPPS activity and a lethal phenotype. PMID- 21643845 TI - An improved particle bombardment for the generation of transgenic plants by direct immobilization of relleasable Tn5 transposases onto gold particles. AB - We have developed a modified particle bombardment method for plant transgenesis. An intein-tag and a 6*Cys-tag were successively fused to the N-terminus of a hyperactive Tn5 transposase. The modified transposase was immobilized on bare gold microscopic particles via covalent binding of a 6*Cys-tag sulfydryl groups to the gold surface. The tethered transposase can bind the transposon DNA in vitro to form the transposome in the absence of Mg2+ ions. After bombardment of the gold particles carrying the transposomes into the plant cells, the transposomes will be released from the carrier due to the activated self-cleavage function of intein-tag. Our data showed this procedure integrated foreign DNA into the plant genome with an increased transformation frequency as compared to the conventional particle bombardment method. A single copy insertion can also be obtained by decreasing of the assembled transposon DNA amount in relation to plant cell biomass. PMID- 21643846 TI - Familial nonsyndromic patent ductus arteriosus caused by mutations in TFAP2B. AB - Patent ductus arteriosus (PDA) is a common congenital heart disease that develops soon after birth when the arterial duct does not remodel. Mutations in TFAP2B, which encodes a neural crest-derived transcription factor, can cause Char syndrome, characterized by PDA, facial dysmorphism, and skeletal abnormalities of the hand. The TFAP2B mutations result in a great amount of phenotypic variability, and a novel TFAP2B mutation has been found in patients with nonsyndromic PDA. Therefore, this study investigated whether TFAP2B mutations can cause familial nonsyndromic PDA. Clinical data and peripheral blood specimens were collected from two kindreds (A and B) and from a cohort of 100 unrelated subjects with PDA. Kindred A spanned three generations, in which 5 of the 16 individuals had PDA, and kindred B spanned three generations, in which 2 of the 13 individuals had PDA. The study enrolled 100 unrelated healthy individuals as control subjects. Polymerase chain reaction (PCR) was used to amplify seven exons and flanking introns of the TFAP2B gene. A few exons of the TFAP2B gene were amplified using reverse transcription polymerase chain reaction (RT-PCR), and direct forward and reverse sequencing of the PCR products was performed. The acquired sequences were aligned with those in GenBank by using a basic local alignment search tool (BLAST). The following two types of mutations were identified in TFAP2B: c.601+5G>A and c.435_438delCCGG. The mutation c.601+5G>A was detected in the affected members of kindred A. Nested PCR showed a splice junction in intron 3 and a 61-bp deletion in exon 3. The mutation c.435_438delCCGG, found in the affected members of kindred B, was caused by a four-base deletion in exon 2, which in turn caused a frame shift that resulted in the formation of a premature stop codon, p.Arg145Argfsx45. None of these mutations was detected in the unaffected members of the kindred or in the control group. Furthermore, two novel single-nucleotide polymorphisms (SNPs), c.1-34G>A and c.539+62G>C, were detected in the introns. The variant c.1-34G>A was identified 34 bp upstream of the transcription initiation site in the TFAP2B gene. Significant differences in the prevalence of the alleles G and A were observed in the control subjects and PDA patients (Z = -2.513, P = 0.012). The study identified that another variant was c.539+62G>C but that the frequency of this variant was similar between the control subjects and the PDA patients (Z = 0.332, P = 0.74). The TFAP2B mutations may be associated with isolated nonsyndromic, hereditary PDA in Chinese families. The authors propose that a TFAP2B mutation should be considered a risk factor for isolated PDA. However, the detailed genetic mechanism underlying nonsyndromic the PDA-causing TFAP2B mutation is yet to be elucidated. PMID- 21643847 TI - Pulmonary limitation to exercise after repair of D-transposition of the great vessels: atrial baffle versus arterial switch. AB - This study evaluated resting pulmonary function and its impact on exercise capacity after atrial baffle (BAFFLE) and arterial switch (SWITCH) repair of D transposition of the great vessels (DTGV). Previously decreased exercise capacity in DTGV patients has been primarily attributed to cardiovascular limitations, whereas pulmonary limitations have largely been overlooked. Resting flow volume loops were compared for BAFFLE (n = 34) and SWITCH (n = 32) patients. Peak exercise variables were compared for BAFFLE (n = 30) and SWITCH (n = 25). Lung disease (restrictive and/or obstructive) was present in 53% of DTGV patients (BAFFLE 62% and SWITCH 44%; p = 0.14). BAFFLE patients had a normal breathing reserve, whereas that of SWITCH patients was decreased (27.3 +/- 28.3 vs. 13.0 +/ 19.2; p = 0.04). BAFFLE patients attained a lower percent of predicted peak oxygen pulse (82.7 +/- 20.5% vs. 94.7 +/- 19.3%; p = 0.04) and peak oxygen consumption (VO(2peak)) (26.6 +/- 6.7 ml/kg/min vs. 37.3 +/- 8.5 ml/kg/min; p < 0.01) than SWITCH patients. Patients after surgical repair for DTGV have an underappreciated occurrence of lung disease, even post-SWITCH. SWITCH patients have diminished breathing reserves, suggesting a pulmonary limitation to VO(2peak). BAFFLE patients have lower VO(2peaks), greater breathing reserves, and lower oxygen pulses than SWITCH patients, suggesting a cardiac limitation to peak aerobic capacity with probable secondary pulmonary limitations. Treating underlying lung disease in symptomatic patients after repair of DTGV may improve functional status. PMID- 21643848 TI - A doctor facing turbulent times: Antoni Tomasz Jurasz, citizen of the world. AB - The eponymous of the Jurasz procedure is Antoni Tomasz Jurasz (1882-1961). The procedure is a standard approach to treating mature pancreatic pseudocysts that are in contact with the stomach, although recent advances in instrumentation have empowered surgeons to perform pseudocystogastrostomy laparoscopically for this problem. Dr. Jurasz was born in Germany but felt himself as Polish as his ancestors. He graduated from Heidelberg with a degree in medicine. Over the following years, he developed a German surgery school, especially helping with the achievements of Erich Lexer and Erwin Payr. During the period between World Wars I and II, Jurasz chaired and led the Surgery Clinic of Poznan University in Poland; and he gave lectures and performed public operations abroad. These activities, together with articles published in German-, French-, and English language medical journals, made him a world-renowned figure in the field of abdominal and thyroid surgery. During World War II, he revealed exceptional traits, taking part in the September Campaign in Poland as a surgeon. He then organized the Polish School of Medicine and the Paderewski Hospital in Edinburgh. Despite his outstanding mind, generosity, and merits, A. T. Jurasz became persona non grata in Communist Poland, which ultimately persuaded him to emigrate to the United States. PMID- 21643849 TI - Regulation of the female rat estrous cycle by a neural cell-specific epidermal growth factor-like repeat domain containing protein, NELL2. AB - NELL2, a protein containing epidermal growth factor-like repeat domains, is predominantly expressed in the nervous system. In the mammalian brain, NELL2 expression is mostly neuronal. Previously we found that NELL2 is involved in the onset of female puberty by regulating the release of gonadotropin-releasing hormone (GnRH), and in normal male sexual behavior by controlling the development of the sexually dimorphic nucleus of the preoptic area (POA). In this study we investigated the effect of NELL2 on the female rat estrous cycle. NELL2 expression in the POA was highest during the proestrous phase. NELL2 mRNA levels in the POA were increased by estrogen treatment in ovariectomized female rats. Blocking NELL2 synthesis in the female rat hypothalamus decreased the expression of kisspeptin 1, an important regulator of the GnRH neuronal apparatus, and resulted in disruption of the estrous cycle at the diestrous phase. These results indicate that NELL2 is involved in the maintenance of the normal female reproductive cycle in mammals. PMID- 21643850 TI - Regulation of Notch1 signaling by Delta-like ligand 1 intracellular domain through physical interaction. AB - Notch signaling involves the proteolytic cleavage of the transmembrane Notch receptor after binding to its transmembrane ligands. The Delta-like ligand 1 also undergoes proteolytic cleavage upon Notch binding, resulting in the production of a free intracellular domain. In this study, we have demonstrated that the Delta like 1 intracellular domain (Dll1-IC) specifically binds to Notch1-IC in the nucleus, thereby disrupting the association of the Notch1-IC-RBP-Jk-MAM transcription activator complex. Additionally, the Notch1-mediated blockage of the induction of MyoD is abolished by the co-expression of Dll1-IC. Collectively, our results show that Dll1-IC functions as a negative regulator in Notch signaling via the disruption of the Notch1-IC-RBP-Jk complex. PMID- 21643851 TI - Isolation of a Leptothrix strain, OUMS1, from ocherous deposits in groundwater. AB - Leptothrix species in aquatic environments produce uniquely shaped hollow microtubules composed of aquatic inorganic and bacterium-derived organic hybrids. Our group termed this biologically derived iron oxide as "biogenous iron oxide (BIOX)". The artificial synthesis of most industrial iron oxides requires massive energy and is costly while BIOX from natural environments is energy and cost effective. The BIOX microtubules could potentially be used as novel industrial functional resources for catalysts, adsorbents and pigments, among others if effective and efficient applications are developed. For these purposes, a reproducible system to regulate bacteria and their BIOX productivity must be established to supply a sufficient amount of BIOX upon industrial demand. However, the bacterial species and the mechanism of BIOX microtubule formation are currently poorly understood. In this study, a novel Leptothrix sp. strain designated OUMS1 was successfully isolated from ocherous deposits in groundwater by testing various culture media and conditions. Morphological and physiological characters and elemental composition were compared with those of the known strain L. cholodnii SP-6 and the differences between these two strains were shown. The successful isolation of OUMS1 led us to establish a basic system to accumulate biological knowledge of Leptothrix and to promote the understanding of the mechanism of microtubule formation. Additional geochemical studies of the OUMS1 related microstructures are expected provide an attractive approach to study the broad industrial application of bacteria-derived iron oxides. PMID- 21643852 TI - Can your dog read your mind?: Understanding the causes of canine perspective taking. AB - Prior studies have documented the domestic dog's (Canis lupus familiaris) sensitivity to human attentional state, including a tendency to preferentially beg for food from attentive individuals and an ability to selectively perform forbidden behaviors when humans are not looking. Due to the success of dogs on perspective-taking tasks, some have hypothesized that domestic dogs may have theory of mind, or the ability to infer what other individuals know. Here we provide the first evidence that nondomesticated canids, grey wolves (Canis lupus), are also sensitive to human attentional state under some conditions. We also demonstrate that dogs do not display an undifferentiated sensitivity to all visual cues of attentional state. Rather, dogs are more sensitive to stimuli encountered in their home environment. Some dogs perform poorly on perspective taking tasks. These findings have important implications for the interpretation of research designed to understand complex social cognition across species. PMID- 21643854 TI - Using GC * GC-FID profiles to estimate the age of weathered gasoline samples. AB - Predicting the amount of time that a petroleum mixture has been exposed to weathering effects has applications in areas of environmental and other forensic investigations, such as aiding in determining the cause and intent of a fire. Historically, research on the evaporation rates of hydrocarbon mixtures has focused on forensic oil spill identification and predicting if a fresh sample could be weathered to give an observed composition in an aged sample. Relatively little attention has focused on approaching the problem from the other direction: estimating exposure time based on the observed composition of a weathered sample at a given time and assuming a prior composition. Here, we build upon our previous research into the weathering of model mixtures by extending our work to gasoline. Samples of gasoline with varying octane ratings and from several vendors were weathered under controlled conditions and their composition monitored over time by two-dimensional gas chromatography (GC * GC). A variety of chemometric models were explored, including partial least squares (PLS), nonlinear PLS (PolyPLS) and locally weighted regression (LWR). A hierarchical application of multivariate techniques was able to predict the time for which a sample had been exposed to evaporative weathering. Partial least squares discriminant analysis could predict whether a sample was relatively fresh (<12 h exposure time) or highly weathered (>20 h exposure time). Subsequent regression models for these classes were evaluated for accuracy using the root mean square error of prediction. LWR was the most successful, whereby fresh and highly weathered samples were predicted to within 30 min and 5 h of exposure, respectively. PMID- 21643853 TI - Effect of acute soman exposure on GABA(A) receptors in rat hippocampal slices and cultured hippocampal neurons. AB - Exposure of the central nervous system to organophosphorus (OP) nerve agents causes seizures and neuronal cell death. Benzodiazepines are commonly used to treat seizures induced by OPs. However, it is known that soman-induced seizures are particularly resistant to benzodiazepine treatment, as compared with other OPs. This study investigated the effect of soman on gamma-aminobutyric acid (GABA) neurotransmission in acute rat hippocampal slices and the surface expression of GABA(A) receptors in cultured rat hippocampal neurons. Results showed that GABA-mediated inhibitory post synaptic currents (IPSCs) are significantly reduced by soman in a concentration-dependent manner in acute rat hippocampal slices. Furthermore, confocal microscopic and cell-based ELISA assays revealed that soman caused rapid internalization of GABA(A) receptors in cultured rat hippocampal neurons. The effect of soman on GABA(A)R endocytosis was not due to inhibition of acetylcholinesterase (AChE) because (1) the acetylcholine muscarinic receptor antagonist atropine did not block soman-induced GABA(A)R endocytosis; and (2) physostigmine, at concentrations that completely inhibit AChE activity, did not cause GABA(A)R endocytosis. Moreover, blocking of the N methyl-D-aspartate (NMDA) receptors by 2-amino-5-phosphonovalerate (APV) had no effect on soman-induced GABA(A)R endocytosis, suggesting that the soman effect was not secondary to glutamate receptor over activation. Regardless of the exact mechanism, the observation that soman induces rapid GABA(A)R endocytosis may have significant implications in the development of effective countermeasures against soman-induced seizures. PMID- 21643855 TI - Quantitative analysis of specific target DNA oligomers using a DNA-immobilized packed-column system. AB - Although a DNA-immobilized packed-column (DNA-packed column), which relies on sequence-dependent interactions of target DNA or mRNA (in the mobile phase) with DNA probes (on the silica particle) in a continuous flow process, could be considered as an alternative platform for quantitative analysis of specific DNA to DNA chip methodology, the performance in practice has not been satisfactory. In this study, we set up a more efficient quantitative analysis system based on a DNA-packed column by employing a temperature-gradient strategy and DMSO containing mobile phase. Using a temperature-gradient strategy based on T(m) values of probe/target DNA hybridizations and DMSO (5%)-containing mobile phase, we succeeded in the quantitative analysis of a specific complementary target distinguishable from non-complementary DNA oligomers or other similar DNA samples. In addition, two different target DNA oligomers even with similar T(m) values were separated and detected quantitatively by using a packed column carrying two different DNA probes. PMID- 21643856 TI - Development and validation of an HPLC-UV detection assay for the determination of rufinamide in human plasma and saliva. AB - The development of a simple and rapid high-performance liquid chromatography (HPLC) method for the determination of the new antiepileptic drug rufinamide (RFN) in human plasma and saliva is reported. Samples (250 MUl) are alkalinized with ammonium hydroxide (pH 9.25) and extracted with dichloromethane using metoclopramide as internal standard. Separation is achieved with a Spherisorb silica column (250 * 4.6 mm i.d., 5 MUm) at 30 degrees C using as mobile phase a solution of methanol/dichloromethane/n-hexane 10/25/65 (vol/vol/vol) mixed with 6 ml ammonium hydroxide. The instrument used was a Shimadzu LC-10Av chromatograph and flow rate was 1.5 ml min(-1), with a LaChrom L-7400 UV detector set at 230 nm. Calibration curves are linear [r(2) = 0.998 +/- 0.002 for plasma (n = 10) and r(2) = 0.999 +/- 0.001 for saliva (n = 9)] over the range of 0.25-20.0 MUg ml( 1), with a limit of quantification at 0.25 MUg ml(-1). Precision and accuracy are within current acceptability standards. The assay is suitable for pharmacokinetic studies in humans and for therapeutic drug monitoring. PMID- 21643857 TI - Segregation of human prostate tissues classified high-risk (UK) versus low-risk (India) for adenocarcinoma using Fourier-transform infrared or Raman microspectroscopy coupled with discriminant analysis. AB - Vibrational spectroscopy techniques can be applied to identify a susceptibility to-adenocarcinoma biochemical signature. A sevenfold difference in incidence of prostate adenocarcinoma (CaP) remains apparent amongst populations of low- (e.g. India) compared with high-risk (e.g. UK) regions, with migrant studies implicating environmental and/or lifestyle/dietary causative factors. This study set out to determine the biospectroscopy-derived spectral differences between risk-associated cohorts to CaP. Benign prostate tissues were obtained using transurethral resection from high-risk (n = 11, UK) and low-risk (n = 14, India) cohorts. Samples were analysed using attenuated total reflection Fourier transform infrared (FTIR) spectroscopy, FTIR microspectroscopy and Raman microspectroscopy. Spectra were subsequently processed within the biochemical cell region (1,800(-1)-500 cm(-1)) employing principal component analysis (PCA) and linear discriminant analysis (LDA) to determine whether wavenumber absorbance/intensity relationships might reveal biochemical differences associated with region-specific susceptibility to CaP. PCA-LDA scores and corresponding cluster vector plots identified pivotal segregating biomarkers as 1,582 cm(-1) (Amide I/II trough); 1,551 cm(-1) (Amide II); 1,667 cm(-1) (Amide I); 1,080 cm(-1) (DNA/RNA); 1,541 cm(-1) (Amide II); 1,468 cm(-1) (protein); 1,232 cm(-1) (DNA); 1,003 cm(-1) (phenylalanine); 1,632 cm(-1) [right-hand side (RHS) Amide I] for glandular epithelium (P < 0.0001) and 1,663 cm(-1) (Amide I); 1,624 cm(-1) (RHS Amide I); 1,126 cm(-1) (RNA); 1,761, 1,782, 1,497 cm(-1) (RHS Amide II); 1,003 cm(-1) (phenylalanine); and 1,624 cm(-1) (RHS Amide I) for adjacent stroma (P < 0.0001). Primarily protein secondary structure variations were biomolecular markers responsible for cohort segregation with DNA alterations exclusively located in the glandular epithelial layers. These biochemical differences may lend vital insights into the aetiology of CaP. PMID- 21643858 TI - Optimization of a miniaturized DBD plasma chip for mercury detection in water samples. AB - In this work, an optimization study was conducted to investigate the performance of a custom-designed miniaturized dielectric barrier discharge (DBD) microplasma chip to be utilized as a radiation source for mercury determination in water samples. The experimental work was implemented by using experimental design, and the results were assessed by applying statistical techniques. The proposed DBD chip was designed and fabricated in a simple way by using a few microscope glass slides aligned together and held by a Perspex chip holder, which proved useful for miniaturization purposes. Argon gas at 75-180 mL/min was used in the experiments as a discharge gas, while AC power in the range 75-175 W at 38 kHz was supplied to the load from a custom-made power source. A UV-visible spectrometer was used, and the spectroscopic parameters were optimized thoroughly and applied in the later analysis. Plasma characteristics were determined theoretically by analysing the recorded spectroscopic data. The estimated electron temperature (T(e) = 0.849 eV) was found to be higher than the excitation temperature (T(exc) = 0.55 eV) and the rotational temperature (T(rot) = 0.064 eV), which indicates non-thermal plasma is generated in the proposed chip. Mercury cold vapour generation experiments were conducted according to experimental plan by examining four parameters (HCl and SnCl(2) concentrations, argon flow rate, and the applied power) and considering the recorded intensity for the mercury line (253.65 nm) as the objective function. Furthermore, an optimization technique and statistical approaches were applied to investigate the individual and interaction effects of the tested parameters on the system performance. The calculated analytical figures of merit (LOD = 2.8 MUg/L and RSD = 3.5%) indicates a reasonable precision system to be adopted as a basis for a miniaturized portable device for mercury detection in water samples. PMID- 21643859 TI - Drug detection in breath: effects of pulmonary blood flow and cardiac output on propofol exhalation. AB - Breath analysis could offer a non-invasive means of intravenous drug monitoring if robust correlations between drug concentrations in breath and blood can be established. In this study, propofol blood and breath concentrations were determined in an animal model under varying physiological conditions. Propofol concentrations in breath were determined by means of two independently calibrated analytical methods: continuous, real-time proton transfer reaction mass spectrometry (PTR-MS) and discontinuous solid-phase micro-extraction coupled with gas chromatography mass spectrometry (SPME-GC-MS). Blood concentrations were determined by means of SPME-GC-MS. Effects of changes in pulmonary blood flow resulting in a decreased cardiac output (CO) and effects of dobutamine administration resulting in an increased CO on propofol breath concentrations and on the correlation between propofol blood and breath concentrations were investigated in seven acutely instrumented pigs. Discontinuous propofol determination in breath by means of alveolar sampling and SPME-GC-MS showed good agreement (R(2)=0.959) with continuous alveolar real-time measurement by means of PTR-MS. In all investigated animals, increasing cardiac output led to a deterioration of the relationship between breath and blood propofol concentrations (R(2)=0.783 for gas chromatography-mass spectrometry and R(2)=0.795 for PTR-MS). Decreasing pulmonary blood flow and cardiac output through banding of the pulmonary artery did not significantly affect the relationship between propofol breath and blood concentrations (R(2)>0.90). Estimation of propofol blood concentrations from exhaled alveolar concentrations seems possible by means of different analytical methods even when cardiac output is decreased. Increases in cardiac output preclude prediction of blood propofol concentration from exhaled concentrations. PMID- 21643860 TI - PCBP2 siRNA reverses the alcohol-induced pro-fibrogenic effects in hepatic stellate cells. AB - PURPOSE: Type I collagen accumulates during liver fibrosis primarily because alpha-complex protein-2 (alphaCP(2)), encoded by the poly(rC) binding protein 2 (PCBP2) gene, binds to the 3' end of the collagen mRNA and increases its half life. This study aimed to reverse the pro-fibrogenic effect of alcohol on hepatic stellate cells (HSCs) by silencing the PCBP2 gene with siRNA. METHODS: The silencing effects of a series of predesigned PCBP2 siRNAs were evaluated in the rat hepatic stellate cell line, HSC-T6. The pro-fibrogenic effects of alcohol on the expression levels of PCBP2 and type-I collagen were examined by several methods. The effect of PCBP2 siRNA on the stability of type I collagen alpha1(I) mRNA was investigated by an in vitro mRNA decay assay. RESULTS: We identified one potent PCBP2 siRNA that reversed the alcohol-induced expression of PCBP2 in HSCs. The decay rate of the collagen alpha1(I) mRNA increased significantly in HSCs treated with the PCBP2 siRNA. CONCLUSION: This study provides the first evidence that alcohol up-regulates the expression of PCBP2, which subsequently increases the half-life of collagen alpha1(I) mRNA. Silencing of PCBP2 using siRNA may provide a promising strategy to reverse the alcohol-induced pro-fibrogenic effects in HSCs. PMID- 21643861 TI - Atypical movement performance and sensory integration in Asperger's syndrome. AB - The aims of this study were to investigate whether individuals with AS have impaired motor abilities and sensorimotor processing and whether these impairments were age-related. Sensorimotor abilities were examined using the Movement Assessment Battery for Children-2, and the Sensory Integration Praxis Test. Fifty boys with AS aged 7-14 years old were compared with typically developing boys. Overall, children with AS showed significant impairment of movement performance as well as proprioceptive and vestibular processing. There were no interaction effects of age and clinical group on level of performance deficit in any of the modalities tested. Increasing our understanding of motor and sensory impairment in AS could have treatment implications for those supporting individuals with AS. PMID- 21643863 TI - Aflatoxin B1 level in relation to child's feeding and growth. AB - OBJECTIVE: To study the influence of sociological factors, breast feeding and weaning on aflatoxin exposure in children as well as to determine the effect of aflatoxin exposure on child's growth. METHODS: A questionnaire, administered to the mothers of forty-six children, obtained information on the child's age, sex, residence, feeding, weaning and general health status. Maternal parity, education and occupation were also collected. Height for age Z-score (HAZ) and weight for age Z-score (WAZ) of children were calculated at the time of recruitment. TLC analysis was performed for aflatoxin B1 level in studied children and their mothers. RESULTS: Aflatoxin B1 was detected in 17 out of 46 (36.96%) of children's serum at a median concentration of 51.61 (30.565-62.795) ppm and in 17 out of 46 (36.96%) of mother's serum at a median concentration of 50 (35.59 84.93) ppm. Aflatoxin B1 level was neither affected by child's age, sex, residence whether rural or urban, maternal age, parity, education nor occupation. Aflatoxin B1 in breastfed patients was significantly lower than in non-breastfed ones (p = 0.034). Weight for age Z-score (WAZ) showed no significant difference between aflatoxin B1 negative and positive cases (p = 0.422) while height for age Z-score (HAZ) was significantly lower in aflatoxin B1 positive compared to negative cases (p = 0.001). A significant positive correlation between aflatoxin B1 in the present cases and their mothers (r = 0.881, p = 0.0001) and a significant negative correlation between aflatoxin B1 in present cases and their height-z-score (HAZ) (r = -0.460, p = 0.001) was detected. CONCLUSIONS: Breast feeding results in lower aflatoxin exposure. Also, a strong association between aflatoxin exposure and impaired child's growth exists. PMID- 21643864 TI - A training set selection strategy for a universal near-infrared quantitative model. AB - The purpose of this article is to propose an empirical solution to the problem of how many clusters of complex samples should be selected to construct the training set for a universal near infrared quantitative model based on the Naes method. The sample spectra were hierarchically classified into clusters by Ward's algorithm and Euclidean distance. If the sample spectra were classified into two clusters, the 1/50 of the largest Heterogeneity value in the cluster with larger variation was set as the threshold to determine the total number of clusters. One sample was then randomly selected from each cluster to construct the training set, and the number of samples in training set equaled the number of clusters. In this study, 98 batches of rifampicin capsules with API contents ranging from 50.1% to 99.4% were studied with this strategy. The root mean square errors of cross validation and prediction were 2.54% and 2.31% for the model for rifampicin capsules, respectively. Then, we evaluated this model in terms of outlier diagnostics, accuracy, precision, and robustness. We also used the strategy of training set sample selection to revalidate the models for cefradine capsules, roxithromycin tablets, and erythromycin ethylsuccinate tablets, and the results were satisfactory. In conclusion, all results showed that this training set sample selection strategy assisted in the quick and accurate construction of quantitative models using near-infrared spectroscopy. PMID- 21643865 TI - Rubus rosaefolius extract as a natural preservative candidate in topical formulations. AB - Even though the synthetic preservatives may offer a high antimicrobial efficacy, they are commonly related to adverse reactions and regarded as having potentially harmful effects caused by chronic consumption. The development of natural preservatives provides a way of reducing the amount of synthetic preservatives normally used in pharmaceutical and cosmetic preparations. In addition, these agents have less toxic effects and represent a possible natural and safer alternative of the preservatives. The purpose of this research was to evaluate the Rubus rosaefolius Smith extract efficiency as a natural preservative in base formulations. Of the extract, 0.2% (w/w) was assayed for its effectiveness of antimicrobial protection in two different base formulations (emulsion and gel). The microbial challenge test was performed following the standard procedures proposed by The United States Pharmacopoeia 33nd, European Pharmacopoeia 6th, Japanese Pharmacopoeia 15th, and the Cosmetics, Toiletries, and Fragrance Association using standardized microorganisms. The results demonstrated that R. rosaefolius extract at the studied concentration reduced the bacterial inocula, satisfying the criterion in all formulations, even though it was not able to present an effective preservative behavior against fungi. Thus, the investigation of new natural substances with preservative properties that could be applied in pharmaceutical and cosmetic products is relevant due to the possibility of substituting or decreasing the concentration of synthetic preservatives, providing a way for the development of safer formulas for the use of consumers. PMID- 21643866 TI - Radiology-led follow-up system for IVC filters: effects on retrieval rates and times. AB - PURPOSE: Successful IVC filter retrieval rates fall with time. Serious complications have been reported following attempts to remove filters after 3-18 months. Failed retrieval may be associated with adverse clinical sequelae. This study explored whether retrieval rates are improved if interventional radiologists organize patient follow-up, rather than relying on the referring clinicians. METHODS: Proactive follow-up of patients who undergo filter placement was implemented in May 2008. At the time of filter placement, a report was issued to the referring consultant notifying them of the advised timeframe for filter retrieval. Clinicians were contacted to arrange retrieval within 30 days. We compared this with our practice for the preceding year. RESULTS: The numbers of filters inserted during the two time periods was similar, as were the numbers of retrieval attempts and the time scale at which they occurred. The rate of successful retrievals increased but not significantly. The major changes were better documentation of filter types and better clinical follow-up. After the change in practice, only one patient was lost to follow-up compared with six the preceding year. CONCLUSIONS: Although there was no significant improvement in retrieval rates, the proactive, radiology-led approach improved follow-up and documentation, ensuring that a clinical decision was made about how long the filter was required and whether retrieval should be attempted and ensuring that patients were not lost to follow-up. PMID- 21643867 TI - Admission diagnoses of children 0-16 years of age hospitalized with influenza. AB - The prompt diagnosis of influenza enables the institution of antiviral therapy and adequate cohorting of patients, but scarce data are available to help clinicians correctly suspect influenza in children at the time of admission. This 16-year retrospective study assessed the main admission diagnoses of 401 children aged <=16 years hospitalized with virologically confirmed influenza. The clinical data were derived from a systematic review of the medical records of the children. Sepsis-like illness was the main reason for admission in 52% of infants aged <6 months and in 7-16% of the older children. Respiratory symptoms accounted for 38% of admissions, and 15% of children were hospitalized due to acute neurologic conditions, primarily febrile convulsions. Wheezing or exacerbation of asthma was the primary reason for admission in 14% of children aged <3 years. No differences were observed in the admission diagnoses between children with influenza A and B infections. The main admission diagnoses vary widely in different age groups of children with influenza, and only a minority of children are hospitalized for respiratory symptoms. The leading role of sepsis-like illness in infants aged <6 months calls for increased efforts to find protective measures against influenza in this age group. PMID- 21643868 TI - Diversity of pneumococcal surface protein A (PspA) and relation to sequence typing in Streptococcus pneumoniae causing invasive disease in Chinese children. AB - The objective of this paper was to investigate the sequence types (STs) and diversity of surface antigen pneumococcal surface protein A (PspA) in 171 invasive Streptococcus pneumoniae isolates from Chinese children. A total of 171 pneumococci isolates were isolated from Chinese children with invasive pneumococcal diseases (IPD) in 11 hospitals between 2006 and 2008. The pneumococci samples were characterized by serotyping, PspA classification, and multilocus sequence typing (MLST). The PspA of these strains could be assigned to two families. The PspA family 2 was the most common (120/171, 70.1%). No PspA family 3 isolates were detected. Family 1 could be subdivided into two clades, with 42 strains in clade 1 and 9 strains in clade 2, and family 2 could be subdivided into clades 3, 4, and 5, which respectively contained 5, 21, and 14 strains. In total, 65 STs were identified, of which ST320 (30/171, 17.5%), ST271 (23/171, 13.5%), and ST876 (18/171, 10.5%) were the most common types. PspA family 2 and family 1 were dominant among pneumococcal clones isolated from Chinese children with invasive disease. The strains with the same ST always presented in the same PspA family. PMID- 21643870 TI - Increased heat shock protein 70 levels in induced sputum and plasma correlate with severity of asthma patients. AB - Damage-associated molecular pattern molecules such as high-mobility group box 1 protein (HMGB1) and heat shock protein 70 (HSP70) have been implicated in the pathogenesis of asthma. The aim of our study was to examine the induced sputum and plasma concentrations of HSP70 in asthmatic patients to determine their relationship with airway obstruction. Thirty-four healthy controls and 56 patients with persistent bronchial asthma matched for gender and age were enrolled in this study. Spirometry measurements were performed before sputum induction. HSP70 levels in induced sputum and plasma were measured using the ELISA Kit. Sputum and plasma concentrations of HSP70 in asthmatics patients were significantly higher than that in control subjects (sputum, (0.88 ng/ml (0.27 1.88 ng/ml) versus 0.42 ng/ml (0.18-0.85 ng/ml), p < 0.001); plasma, (0.46 ng/ml (0.20-0.98 ng/ml) versus 0.14 ng/ml (0.11-0.37 ng/ml), p < 0.001) and were significantly negatively correlated with forced expiratory volume in 1 s (FEV1), FEV1 (percent predicted), and FEV1/FVC in all 90 participants and 56 patients with asthma. There were no significant differences in HSP70 levels between patients with eosinophilic and non-eosinophilic asthma. HSP70 levels in plasma were positively correlated with neutrophil count, and HSP70 levels in induced sputum were positively correlated with lymphocyte count. In multivariate analysis, independent predictors of sputum HSP70 were diseases and disease severity but not smoking, age, or gender, and independent predictors of plasma HSP70 were also diseases and disease severity. In conclusion, this study indicates that induced sputum and plasma HSP70 could serve as a useful marker for assessing the degree of airway obstruction in patients with asthma. However, further investigation is needed to establish the role of circulating and sputum HSP70 in the pathogenesis of asthma. PMID- 21643871 TI - Avoid hypotension and hypoxia: an old anesthetic adage with renewed relevance from cerebral oximetry monitoring. PMID- 21643872 TI - Academic fraud: perspectives from a lifelong anesthesia researcher. PMID- 21643873 TI - Resolution of pulmonary edema with variable mechanical ventilation in a porcine model of acute lung injury. AB - OBJECTIVE: Resolution of the acute respiratory distress syndrome (ARDS) requires clearance of pulmonary edema. Biologically variable ventilation (BVV) strategies that improve gas exchange, lung mechanics, and inflammatory mediators in ARDS may be beneficial in this regard. We used quantitative computed tomography (CT), a single indicator thermodilution system (PiCCO(r)) to determine extravascular lung water (EVLW), and the change in edema protein concentration over time to compare edema clearance with BVV vs conventional mechanical ventilation (CMV) in a porcine ARDS model. METHODS: Sixteen pigs with oleic acid lung injury were randomized to four hours of ventilation with either CMV (n = 8) or BVV (n = 8) at identical low tidal volume and minute ventilation over time. Hemodynamic variables, gas exchange, lung mechanics, and PiCCO derived EVLW were determined hourly. Computed tomography images and edema fluid samples were obtained at baseline lung injury and after four hours of ventilation. Wet and dry lung weights were determined postmortem. RESULTS: At four hours with BVV, peak airway pressure was decreased significantly and lung compliance improved compared with CMV (P = 0.003; P < 0.001, respectively). Hemodynamic variables and gas exchange were not different between groups. Also at four hours, computed tomography revealed an increase in total gas volume (P = 0.001) and a decrease in total lung weight and global lung density (P = 0.005; P = 0.04 respectively) with BVV. These findings were associated with a significant increase in the gas volume of normally aerated lung regions (P < 0.001) and a decrease in the poorly and non aerated lung regions (P = 0.001). No change in any CT parameter occurred with CMV. The lung weights derived from computed tomography correlated well with postmortem wet weights (R(2) = 0.79; P < 0.01). The decrease in PiCCO derived EVLW from injury to four hours did not differ significantly between BVV and CMV. Extravascular lung water showed no correlation with postmortem wet weights and significantly underestimated lung water. Average alveolar fluid clearance rates were positive (1.4%.hr(-1) (3%)) with BVV and negative with CMV (-2.0%.hr(-1) (4%)). CONCLUSIONS: In a comparison between BVV and CMV, computed tomography evidence suggests that BVV facilitates enhanced clearance and/or redistribution of edema fluid with improved recruitment of atelectatic and poorly aerated lung regions; no such evidence was seen with either single thermodilution measurement of EVLW or edema clearance rates. The results of computed tomography provide further evidence of the benefit of BVV over conventional ventilation in ARDS. PMID- 21643874 TI - Verification of the psychometric properties of the Japanese version of the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire Core 15 palliative (EORTCQLQ-C15-PAL). AB - PURPOSE: We compared Japanese versions of the EORTC QLQ-C15-PAL and QLQ-C30 to assess the utility of the former survey for terminal-phase cancer patients. METHODS: We used QLQ-C30 scores from the survey prior to each patient's death and extracted QLQ-C15-PAL data. We determined intraclass correlation coefficients (ICCs) of the five QLQ-C15-PAL subscales: physical functioning, emotional functioning, fatigue, nausea and vomiting, and global health status/quality of life (QOL) and assessed equivalent-form reliability. Regression analysis was used to determine the extent to which QLQ-C15-PAL items explained the QLQ-C30 score. RESULTS: Our study included 32 of 91 cancer patients receiving palliative care who could self-administer the questionnaire. The ICC between the QLQ-C15-PAL and QLQ-C30 scores was 0.93 or higher. The proportion of variance (R-squared) for each subscale was 0.87 or higher. CONCLUSION: We examined the validity and reliability of the Japanese version of the QLQ-C15-PAL. We found an 87% or higher chance that the QLQ-C15-PAL could explain the original QLQ-C30 score. Therefore, QLQ-C15-PAL appears to be useful for assessing the QOL of terminal-phase cancer patients. PMID- 21643875 TI - The psychosocial impact of cancer: evidence in support of independent general positive and negative components. AB - PURPOSE: Considerable research has demonstrated the negative psychosocial impact of cancer. Recent work has highlighted positive psychosocial outcomes. Research is now needed to evaluate the relationship between negative and positive impacts. This paper reports the development and validation of a measurement model capturing positive and negative psychosocial illness impacts. METHODS: The sample included 754 cancer patients on- or post-treatment. Item development was informed by literature review, expert input patient interviews and the results of a pilot study of 205 cancer patients, resulting in 43 positive and 46 negative items. Factor analyses were used to evaluate the dimensionality of the item pools. Analysis of variance (ANOVA) was used to examine relationships between psychosocial illness impact and other variables. RESULTS: Unidimensionality was demonstrated within but not across negative and positive impact items. ANOVA results showed differential relationships between negative and positive impacts, respectively, and patient sociodemographic and clinical variables. CONCLUSION: Positive and negative psychosocial illness impacts are best conceptualized and measured as two independent factors. Computerized adaptive tests and short-form measures developed from this comprehensive psychosocial illness impact item bank may benefit future research and clinical applications. PMID- 21643876 TI - Subpectoral-subfascial breast augmentation for thin-skinned patients. AB - BACKGROUND: Among thin patients, upper pole skin of less than 2 cm on a pinch test is an indication for submuscular implant placement. However, rippling, palpability, and visibility of the implant are frequently observed in the lower and lateral aspects of the breast due to the lack of muscle coverage in the area. To overcome these problems, a subpectoral-subfascial breast augmentation was performed including subfascial dissection of the rectus abdominis, external abdominal oblique muscle, and serratus anterior muscle as one sheet to cover the inferolateral aspect of the implant. METHODS: From March 2008 to March 2010, 62 thin patients with hypomastia underwent subpectoral-subfascial breast augmentation. For all these patients, the result of the pinch test was less than 2 cm on the upper pole of the breast and less than 1 cm on the lower and lateral aspect. With a transareolar-perinipple incision, pockets were created such that the superomedial portion was covered by the pectoralis major muscle, and the inferolateral portion was covered by the fascial flap of neighboring muscles. RESULTS: The results were satisfactory, reproducible, and predictable. The implants were not palpable or visible during a mean follow-up period of 19 months due to the inclusion of a fascial flap in all the patients. CONCLUSIONS: This technique, which uses the advantages of both subpectoral and subfascial techniques, can offer a high-grade result. Especially for thin patients with less subcutaneous tissue, it provides excellent upper and lower pole coverage and gives the shape of the breast a natural appearance, with no palpability or rippling. PMID- 21643877 TI - Development of a feasible transrectal natural orifice transluminal endoscopic surgery (NOTES(r)) approach in a cadaveric appendectomy model: anterior is better. AB - BACKGROUND: The transrectal natural orifice transluminal endoscopic surgery (NOTES) approach is a potentially promising alternative to transgastric or transvaginal approaches for intraperitoneal procedures. However, whether the optimal transrectal approach for intraperitoneal surgery is anterior or posterior remains unknown. To evaluate this, a prospective comparison of anterior and posterior transrectal NOTES approaches in a cadaveric appendectomy model was performed. METHODS: Operations were performed on human cadavers using a transanal endoscopic microsurgery (TEM) scope to assist with access and closure. Posterior access was achieved by tunneling cephalad through the retrorectal space into the peritoneal cavity. Anterior transrectal access was established through the rectal wall just above the peritoneal reflection. A dual-channel flexible endoscope was used to perform appendectomies. Rectotomies were closed using sutures or staples. Operative time, degree of laparoscopic assistance, complications, and leak testing results were recorded. RESULTS: This study investigated 10 cadavers with access and closure attempted using both anterior (n = 10) and posterior (n = 5) approaches, whereas appendectomies were performed using either an anterior (n = 8) or a posterior (n = 2) approach. The anterior approach required less time than the posterior approach for peritoneal access (4 +/- 1 vs. 61 +/- 14 min; p < 0.001), specimen extraction (2 +/- 1 vs. 5 +/- 1 min; p < 0.01), and the total operation (99 +/- 35 vs. 176 +/- 26 min; p = 0.02). A "pure" NOTES dissection was possible with the anterior approach using rigid transanal instruments for assistance. Dissection time, closure time, and the incidence of complications were similar between the two approaches. Leak testing of closures showed significant variability for all closure types. CONCLUSION: Transrectal NOTES appendectomy is feasible in a cadaveric model using an anterior transrectal approach. This approach is technically easier, results in shorter operative times, and allows for a "pure" NOTES dissection compared with a posterior transrectal approach. Leak pressure testing of NOTES closures is unreliable in the cadaveric model. PMID- 21643878 TI - Endoscopic assessment of failed fundoplication: a case for standardization. AB - BACKGROUND: Preoperative endoscopic assessment of the failed fundoplication is instrumental in diagnosis and surgical management. Endoscopy is a routine and essential part of the workup for a failed fundoplication, but no clear guidelines exist for reporting endoscopic findings. This study aimed to compare endoscopic findings reported by community physicians (gastroenterologists and surgeons) with the findings of the authors (esophageal center) for patients who underwent reoperative intervention after a previous antireflux procedure. METHODS: Retrospective review of a prospectively maintained database was performed to identify patients who underwent reoperation after a failed antireflux operation between 1 December 2003 and 30 June 2010. Endoscopic findings as reported by the outside physician and by the esophageal center endoscopist were reviewed and compared. RESULTS: During the study period, 229 patients underwent reoperation. Of these patients, 20 did not have endoscopy performed by an outside physician and were excluded from the study, leaving 208 patients. The endoscopic reports of the esophageal center physician included 97 cases of hiatal hernia (64 type 1 and 33 types 2 and 3), 52 slipped fundoplications, 61 disrupted fundoplications, 30 intrathoracic fundoplications, 25 twisted fundoplications, 14 two-compartment stomachs, and 27 cases of Barrett's esophagus. Outside physicians identified 68% of the hiatal hernias and 61% of the paraesophageal hernias reported by the authors. Only 32% of the outside reports mentioned a previous fundoplication. Furthermore, only 17% of the slipped fundoplications and 30% of the disrupted fundoplications were so described. Outside physicians identified 19 of the 27 patients with Barrett's esophagus. CONCLUSION: Fundoplication changes described by the general endoscopist are inadequate. With an increasing population of patients who have undergone prior antireflux surgery, incorporation of fundoplication assessment in an endoscopic curriculum may be helpful. PMID- 21643879 TI - Endoscopic fixation of the rectum for rectal prolapse: a feasibility and survival experimental study. AB - BACKGROUND: In recent years, there has been considerable interest in developing technology as well as techniques that could widen the therapeutic horizons of endoscopy. Rectal prolapse, a benign localized condition causing considerable morbidity, could be an excellent focus for new endoscopic therapies. The aim of this study was to assess the feasibility and safety of endoluminal fixation of the rectum to the anterior abdominal wall, after pushing it up inside the body, using an in vivo animal model. METHODS: We performed an in vivo comparative surgical study in a porcine model, including laparoscopic mobilization of the rectum and posterior rectopexy (standard surgical method) or endoluminal tacking of the rectum. After proving feasibility in ex vivo and acute studies, we performed a survival study to evaluate the safety of endoluminal tacking of the mobilized rectum to the anterior abdominal wall. The main outcome measures were successful completion of the tasks, maintenance of the fixation, complications associated with the methods, and survival studies including histopathological examinations of the fixation sites. RESULTS: There were two groups: laparoscopic rectopexy (8 animals) and endoluminal fixation of the rectum to the anterior abdominal wall (10 animals). There were no differences between these two groups in their postoperative recovery. The group with the endoluminal fixation was found to have adequate attachment of the rectum to the anterior abdominal wall (measured attachment pressure in the endoluminal group = 6.06 +/- 0.52 ft-lb, in the control group = 4.86 +/- 2.00 ft-lb) on both gross and microscopic evaluation. CONCLUSION: Endoscopic fixation of the mobilized rectum is feasible and safe in this model and in the future may provide an effective alternative to current treatment options for rectal prolapse. PMID- 21643880 TI - Low risk of trocar site hernia repair 12 years after primary laparoscopic surgery. AB - BACKGROUND: The risk of trocar site hernia (TSH) may be 0-22%, but no large scaled data with long-term follow-up are available. The purpose of this study was to estimate the long-term risk of TSH repair. METHODS: All patients in Denmark who underwent a laparoscopic procedure in 1997 were identified using the Danish National Patient Register and followed during a 12-year period. Hospital records for patients with an incisional or umbilical hernia repair were tracked and manually analyzed for possible relationship between reoperation and the initial laparoscopy. RESULTS: We included 7,626 patients. During follow-up, we identified 95 patients with TSH repair with a cumulative risk of 1.3% being lowest after minor gynecological procedures and appendectomy and highest after fundoplication, cholecystectomy, and oophorectomy. The TSH was mainly at the umbilicus site (n = 63, 66%), and 15 (16%) of the TSH repairs were performed as an emergency procedure. CONCLUSIONS: The long-term risk of TSH repair is low, but the risk of an emergency operation for TSH is relatively high, which suggests that all patients with a TSH should be offered elective repair. PMID- 21643881 TI - Intensive laparoscopic training course for surgical residents: program description, initial results, and requirements. AB - INTRODUCTION: The Department of Surgery at the University of Arizona has created an intensive laparoscopic training course for surgical residents featuring a combined simulation laboratory and live swine model. We herein report the essential components to design and implement a rigorous training course for developing laparoscopic skills in surgical residents. MATERIALS AND METHODS: At our institution, we developed a week-long pilot intensive laparoscopic training course. Six surgical residents (ranging from interns to chief residents) participate in the structured, multimodality course, without any clinical responsibilities. It consists of didactic instruction, laboratory training, practice in the simulation laboratory, and performance (under the direction of attending laparoscopic surgeons) of surgical procedures on pigs. The pigs are anesthetized and attended by veterinarians and technicians, and then euthanized at the end of each day. Three teams of two different training-level residents are paired. Daily briefing, debriefing, and analysis are performed at the close of each session. A written paper survey is completed at the end of the course. RESULTS: This report describes the results of first 36 surgical residents trained in six courses. Preliminary data reveal that all 36 now feel more comfortable handling laparoscopic instruments and positioning trocars; they now perform laparoscopic surgery with greater confidence and favor having the course as part of their educational curriculum. CONCLUSION: A multimodality intensive laparoscopic training course should become a standard requirement for surgical residents, enabling them to acquire basic and advanced laparoscopic skills on a routine basis. PMID- 21643882 TI - Impact of pressure and gas type on adhesion formation and biomaterial integration in laparoscopy. AB - BACKGROUND: Laparoscopic mesh repair of inguinal and incisional hernias has been widely adopted. Nevertheless, knowledge about the impact of pneumoperitoneum on mesh integration is rare. The present study investigates pressure and gas dependent effects of pneumoperitoneum on adhesion formation and biomaterial integration in a standardized animal model. METHODS: Laparoscopic intraperitoneal onlay mesh implantation (IPOM) was performed in 32 female chinchilla rabbits using CO(2) or helium for pneumoperitoneum. Intra-abdominal pressures were 3 or 6 mmHg. Animals were killed after 21 days, and the abdominal wall was explanted for subsequent histopathological examinations. Adhesions were assessed qualitatively with a scoring system, and the adhesion surface was analyzed semiquantitatively by planimetry. Infiltration of macrophages (CD68), expression of matrix metalloproteinase 13 (MMP-13), and cell proliferation (Ki67) were analyzed at the mesh to host interface by immunohistochemistry. The collagen type I/III ratio was analyzed by cross-polarization microscopy to determine the quality of mesh integration. RESULTS: After 21 days, perifilamental infiltration with macrophages (CD68) and percentage of proliferating cells (Ki67) were highest after 6 mmHg of CO(2) pneumoperitoneum. The extent of adhesions, as well as the expression of MMP 13 and the collagen type I/III ratio, were similar between groups. CONCLUSIONS: Our experiments showed no pressure or gas-dependent alterations of adhesion formation and only minor effects on biomaterial integration. Altogether, there is no evidence for a clinically negative effect of CO(2) pneumoperitoneum. PMID- 21643883 TI - Tempo and the TFR. AB - Tempo effects in period fertility indicators are widely regarded as a source of bias or distortion. But is this always the case? Whether tempo change results in bias depends, in the view advanced here, on the measure used, the meaning of bias/distortion, and the objective of analysis. Two ways of construing bias in period measures are suggested, and their relevance is discussed in the context of five broad purposes for measuring period fertility: describing and explaining fertility time trends, anticipating future prospects, providing input parameters for formal models, and communicating with nonspecialist audiences. Genuine timing effects are not biasing when period fertility is the explanandum but are distorting when the aim is to estimate cohort fertility. Alternatives to tempo adjustment are available that are a more defensible solution to the issue of timing change. Tempo adjustment could be more fruitfully considered a form of modeling rather than empirical measurement. The measurement of period fertility could be improved by relying more on a statistical approach and less on indicators based on stable assumptions. Future progress will depend on integrating research on measurement with substantive investigation. PMID- 21643884 TI - Mortgage foreclosure and health disparities: serial displacement as asset extraction in African American populations. AB - In this paper we offer a conceptualization of mortgage foreclosure as serial displacement by highlighting the current crisis in the context of historically repeated extraction of capital-economic, social, and human-from communities defined at different scales: geographically, socially, and that of embodied individuals. We argue that serial displacement is the loss of capital, physical resources, social integration and collective capacity, and psycho-social resources at each of these scales, with losses at one level affecting other levels. The repeated extraction of resources has negative implications for the health of individuals and groups, within generations as well as across generations, through the accumulation of loss over time. Our analysis of the foreclosure crisis as serial displacement for African American households in the United States begins with the "housing niche" model. We focus on the foreclosure crisis as an example of the interconnectedness of structured inequality in health and housing. Then we briefly review the history of policies related to racial inequality in homeownership in the twentieth and twenty-first centuries. We end with an analysis of the scales of displacement and the human, social, and capital asset extraction that accompany them. PMID- 21643885 TI - Oblique axial MR imaging of the normal anterior cruciate ligament bundles. PMID- 21643886 TI - Radiological features of Paget disease of bone associated with VCP myopathy. AB - OBJECTIVE: Mutations in the Valosin-containing protein (VCP) gene cause a unique disorder characterized by classic Paget disease of bone (PDB), inclusion body myopathy, and frontotemporal dementia (IBMPFD). Our objective was to analyze the radiographic features of PDB associated with VCP mutations since there is a dearth of literature on the PDB component of VCP disease. MATERIALS AND METHODS: Radiographic bone surveys were examined in 23 individuals with VCP mutation and compared with their unaffected relatives. Laboratory testing relevant for VCP disease was performed in all individuals. RESULTS: Of the 17 affected individuals with clinical manifestations of VCP disease, 16 of whom had myopathy, radiographic analysis revealed classic PDB in 11 individuals (65%). The mean age of diagnosis for myopathy was 43.8 years and for PDB was 38.1 years of age. Radiological evidence of PDB was seen in one individual (16%) amongst six clinically asymptomatic VCP mutation carriers. Alkaline phosphatase was a useful marker for diagnosing PDB in VCP disease. CONCLUSIONS: Radiographic findings of classic PDB are seen in 52% of individuals carrying VCP mutations at a significantly younger age than conventional PDB. Screening for PDB is warranted in at-risk individuals because of the benefit of early treatment with the new powerful bisphosphonates that hold the potential for prevention of disease. PMID- 21643887 TI - Effect of radiologist experience on the risk of false-positive results in breast cancer screening programs. AB - OBJECTIVES: To evaluate the effect of radiologist experience on the risk of false positive results in population-based breast cancer screening programmes. METHODS: We evaluated 1,440,384 single-read screening mammograms, corresponding to 471,112 women aged 45-69 years participating in four Spanish programmes between 1990 and 2006. The mammograms were interpreted by 72 radiologists. RESULTS: The overall percentage of false-positive results was 5.85% and that for false-positives resulting in an invasive procedure was 0.38%. Both the risk of false-positives overall and of false-positives leading to an invasive procedure significantly decreased (p < 0.001) with greater reading volume in the previous year: OR 0.77 and OR 0.78, respectively, for a reading volume 500-1,999 mammograms and OR 0.59 and OR 0.60 for a reading volume of >14,999 mammograms with respect to the reference category (<500). The risk of both categories of false-positives was also significantly reduced (p < 0.001) as radiologists' years of experience increased: OR 0.96 and OR 0.84, respectively, for 1 year's experience and OR 0.72 and OR 0.73, respectively, for more than 4 years' experience with regard to the category of <1 year's experience. CONCLUSION: Radiologist experience is a determining factor in the risk of a false-positive result in breast cancer screening. PMID- 21643888 TI - Ectopic overexpression of vacuolar and apoplastic Catharanthus roseus peroxidases confers differential tolerance to salt and dehydration stress in transgenic tobacco. AB - CrPrx and CrPrx1 are class III peroxidases previously cloned and characterized from Catharanthus roseus. CrPrx is known to be apoplastic in nature, while CrPrx1 is targeted to vacuoles. In order to study their role in planta, these two peroxidases were expressed in Nicotiana tabacum. The transformed plants exhibited increased peroxidase activity. Increased oxidative stress tolerance was also observed in transgenics when treated with H(2)O(2) under strong light conditions. However, differential tolerance to salt and dehydration stress was observed during germination of T1 transgenic seeds. Under these stresses, the seed germination of CrPrx-transformed plants and wild-type plants was clearly suppressed, whereas CrPrx1 transgenic lines showed improved germination. CrPrx transformed lines exhibited better cold tolerance than CrPrx1-transformed lines. These results indicate that vacuolar peroxidase plays an important role in salt and dehydration stress over cell wall-targeted peroxidase, while cell wall targeted peroxidase renders cold stress tolerance. PMID- 21643889 TI - From molecules to myofibers: multiscale imaging of the myocardium. AB - Pathology in the heart can be examined at several scales, ranging from the molecular to the macroscopic. Traditionally, fluorescence-based techniques such as flow cytometry have been used to study the myocardium at the molecular, cellular, and microscopic levels. Recent advances in magnetic resonance imaging (MRI), however, have made it possible to image certain cellular and molecular events in the myocardium noninvasively in vivo. In addition, diffusion MRI has been used to image myocardial fiber architecture and microstructure in the intact heart. Diffusion MRI tractography, in particular, is providing novel insights into myocardial microsctructure in both health and disease. Recent developments have also been made in fluorescence imaging, making it possible to image fluorescent probes in the heart of small animals noninvasively in vivo. Moreover, techniques have been developed to perform in vivo fluorescence tomography of the mouse heart. These advances in MRI and fluorescence imaging allow events in the myocardium to be imaged at several scales linking molecular changes to alterations in microstructure and microstructural changes to gross function. A complete and integrated picture of pathophysiology in the myocardium is thus obtained. This multiscale approach has the potential to be of significant value not only in preclinical research but, ultimately, in the clinical arena as well. PMID- 21643890 TI - Baseline immune phenotypes and CD4+ T lymphocyte responses to antiretroviral therapy in younger versus older HIV-infected individuals. AB - OBJECTIVE: The purpose of the study was to determine associations between pre antiretroviral therapy (ART) senescent CD8+ T lymphocytes and naive versus non naive CD8+ and CD4+ T lymphocyte subpopulations and CD4+ responses after initiation of ART in younger versus older individuals. METHODS: Retrospective analysis of 100 subjects with pre-ART cryopreserved peripheral blood mononuclear cells samples was performed with flow cytometry. Subjects were divided into four groups by age (30-50 years or > 50 years) and 96-week CD4+ response (<100 or >200 cells/mm(3)). All subjects had 96-week viral suppression to <50 copies/mm(3). Regression was utilized to investigate associations between pre-ART CD8+ and CD4+ T cell phenotypes with age and CD4+ response categories. RESULTS: Individuals <50 years had a lower frequency of senescent CD8+ T lymphocytes of the CD56 + 57+, CD56+, and CD28- phenotypes (95%CI -3.6 to -0.02; 95%CI -4.2 to -0.03; 95%CI 12.5 to -1.4, respectively) and a higher frequency of naive (CD45RA + CD28+) CD8+ T lymphocytes (95%CI 2.6 to 10.9). Younger age and good CD4+ response were associated with a higher frequency of pre-ART naive CD4+ T cells (95%CI 2.0 to 16.4 and 95%CI 1.5 to 15.6, respectively). CONCLUSIONS: Prior to ART, younger HIV infected individuals have a higher frequency of naive CD4+ and CD8+ T cells and lower frequency of senescent CD8+ T cell phenotypes. PMID- 21643891 TI - The relationship between hypogammaglobulinemia, monoclonal gammopathy of undetermined significance and humoral immunodeficiency: a case series. AB - Hypogammaglobulinemia of the non-monoclonal immunoglobulin heavy chain classes has been reported in monoclonal gammopathy of undetermined significance (MGUS) patients. Whether low polyclonal immunoglobulin levels are associated with impaired specific antibody production and whether they represent a risk factor for the development of recurrent bacterial infections have not been established in this population. We determined the frequency of MGUS in patients referred to a tertiary care clinical immunology ambulatory care practice for evaluation of hypogammaglobulinemia, who were assessed for deficits in specific antibody production and the presence of recurrent infections. Of the 133 patients evaluated for hypogammaglobulinemia, 68 were screened for monoclonal gammopathy and 5 were found to have MGUS. Three had MGUS associated hypogammaglobulinemia in the absence of a defining primary immunodeficiency, one possibly had common variable immunodeficiency, and one had an uncertain diagnosis. Thus, MGUS may be uncovered in patients presenting with hypogammaglobulinemia even in those who lack an elevated serum level of IgG, IgM, or IgA. PMID- 21643892 TI - Incidence of infection is inversely related to steady-state (trough) serum IgG level in studies of subcutaneous IgG in PIDD. AB - Results from seven studies of four subcutaneous IgG preparations in patients with primary immune deficiencies show that the incidence of infection is inversely related to the steady-state IgG level. Maintaining higher IgG levels may be beneficial, and no given level is necessarily adequate for all patients. PMID- 21643893 TI - Dehydroepiandrosterone suppresses eosinophil infiltration and airway hyperresponsiveness via modulation of chemokines and Th2 cytokines in ovalbumin sensitized mice. AB - In this study, we evaluated the anti-inflammatory response and the mechanism by which dehydroepiandrosterone modulates immunity in ovalbumin-sensitized asthmatic mice. Female BALB/c mice were sensitized and challenged with ovalbumin and then treated with oral administration of dehydroepiandrosterone on days 21 to 27. The results showed dehydroepiandrosterone could suppress airway hyperresponsiveness and decrease eosinophil infiltration of the lungs in ovalbumin-sensitized mice. Moreover, dehydroepiandrosterone inhibited chemokines, including CCL11/eotaxin-1 and CCL24/eotaxin-2, and Th2-associated cytokine levels in bronchoalveolar lavage fluid. After the inflammatory human bronchial epithelial cell line BEAS-2B was treated with dehydroepiandrosterone, levels of proinflammatory cytokines and chemokines were inhibited, including IL-6, IL-8, CCL11, and CCL24. We suggested that dehydroepiandrosterone inhibited inflammation in bronchial epithelial cells as indicated by the suppression of Th2-associated cytokines and chemokines. Dehydroepiandrosterone also suppressed eosinophil migration and infiltration into the lung to improve the symptoms of asthma in ovalbumin-sensitized mice. PMID- 21643894 TI - Putting in shape: towards a unified approach for the taxonomic description of monogenean haptoral hard parts. AB - Among monogeneans, haptoral hard parts provide prominent morphological characters upon which identifications are largely based. Traditionally, morphometric approaches are based on the use of arbitrary collections of linear distance measurements between landmarks. An exhaustive review of the specific diagnoses published in the journal Systematic Parasitology highlights the fact that an intricately important number of measurements are used to describe the same morphological features. Hence, this does not allow relevant comparison between studies and may have caused confusion in the literature. More importantly, a significant proportion of diagnoses commonly used do not maximize the amount of information available from morphological features, and sets of linear measurements between landmarks do not properly allow the complete reconstruction of the shape of haptoral hard parts. Given this prominent bias and the disparate use of traditional methodologies, I suggest the use of alternative methods in systematic parasitology that fully take into consideration the shape of morphological features. In addition to these considerations, a move toward placing shape at the centre of automated species recognition would be mutually beneficial for both taxonomists and non-taxonomists. PMID- 21643895 TI - Two new species of parasitic copepods (Crustacea) on two immigrant rabbitfishes (Family Siganidae) from the Red Sea. AB - Two new species of parasitic copepods, one from each of the families Hatschekiidae and Bomolochidae, are reported from two immigrant species of rabbitfishes (Family Siganidae), both of which originated from the Red Sea but are now established in the Mediterranean. The descriptions of Hatschekia siganicola n. sp. and Nothobomolochus neomediterraneus n. sp. are based on material of both sexes obtained from the gills of Siganus luridus Ruppell and S. rivulatus Forsskal, respectively, caught in Egyptian Mediterranean waters off the Alexandrian coast. PMID- 21643896 TI - Hexamermis eurygasteri n. sp. (Nematoda: Mermithidae) parasitising the sunn pest Eurygaster integriceps Puton (Hemiptera: Scutelleridae) in Turkey. AB - A new species of mermithid nematode, Hexamermis eurygasteri n. sp. (Nematoda: Mermithidae) is described as a parasite of the sunn pest Eurygaster integriceps Puton (Hemiptera: Scutelleridae) in Turkey. The combination of the following characters separate H. eurygasteri from other members of Hexamermis Steiner, 1924, as defined by Artyukhovsky (1990) and Kaiser (1991): amphidial pouch integrated into lateral cephalic papillae; amphidial openings minute; well developed cuticular vulval cone; small vulval lips; vagina straight or slightly curved at tip, without reverse bend; spicules shorter in length than body diameter at cloaca; spicules straight, except occasionally for short bend at base; spicule tips bluntly rounded; and two double rows of genital papillae. This is the first description of a nematode parasite of a member of the Scutelleridae and the first description of a mermithid nematode from Turkey. This mermithid has potential as a biological control agent in an integrated control programme of the sunn pest. PMID- 21643897 TI - Further investigations of the mite genus Syringophiloidus Kethley, 1970 (Acariformes: Syringophilidae) from North American passerines. AB - Four new syringophilid species of Syringophiloidus Kethley, 1970 are described from North American passerines: S. zonotrichia n. sp. from Zonotrichia albicolis (Gmelin) (Emberizidae) on Texas; S. jackowiaki n. sp. from Poecile carolinensis (Auduborn) (Paridae) in Texas; and S. xanthocephalus n. sp. from Xanthocephalus xanthocephalus (Bonaparte) (Icteridae) and S. agelaius n. sp. from Agelaius phoeniceus Linnaeus (Icteridae), both from Arizona. Spizella breweri (Cassin) (Emberizidae) from California is a new host for Syringophiloidus sialius Skoracki, Flannery & Spicer, 2009; and Melospiza lincolnii (Auduborn) (Emberizidae) from Texas and Vermivora ruficapilla (Wilson) (Parulidae) from California are new hosts for S. seiuri (Ckark, 1964). S. daberti Bochkov, Fain & Skoracki, 2004 from Passerina ciris Linnaeus (Cardinalidae) is recorded in the USA for the first time. A table with the host associations and distribution of all of the North American species of Syringophiloidus is given. PMID- 21643898 TI - Species of Haliotrema Johnston & Tiegs, 1922 (Monogenea: Dactylogyridae) from Zanclus cornutus (L.) (Teleostei: Zanclidae) and Acanthurus nigrofuscus (Forsskal) (Teleostei: Acanthuridae) in the South China Sea. AB - Four species of Haliotrema Johnston & Tiegs, 1922, including three new taxa, are described from the gills of two species of coral reef fishes, Zanclus cornutus (Linnaeus) and Acanthurus nigrofuscus (Forsskal), off the Dongsha Islands in the South China Sea. Haliotrema dongshaense n. sp., from Z. cornutus, is differentiated from other existing congeneric species by its peculiar male copulatory organ, comprising a harp-shaped copulatory tube and a cup-shaped base, and two groups of short longitudinal muscles lying on either side of the vaginal vestibule. Haliotrema zigmoidocirrus n. sp. from Z. cornutus and H. nigrofusci n. sp. from A. nigrofuscus are differentiated from other congeneric species by their male copulatory organ, which has a cup-shaped base, bell-shaped middle and Z shaped distal part, and the latter can be readily differentiated from the former by its distinctively wider haptor and longer connecting bars. Haliotrema sicklocirrus Wang, 2007, from Z. cornutus, is redescribed with additional details, including the sinistral position of the accessory piece of the male copulatory organ, the absence of eyespots and the morphology of the connecting bars. PMID- 21643899 TI - Four new xarifiid copepods (Poecilostomatoida) associated with the scleractinian coral Pavona explanulata (Lamarck) from off Taiwan. AB - Four new xarifiid copepods are described. They were found in association with the scleractinian coral Pavona explanulata (Lamarck) occurring in shallow water reefs off Yenliao in northern Taiwan. The four species are: Xarifia capillata n. sp., X. parva n. sp., X. pavonae n. sp. and X. taiwanensis n. sp. They were found together in a single washing of the host coral. Previously, 13 species of copepods have been found in association with nine species of Pavona Lamarck. More than half (7/13) of these symbionts are members of Xarifia Humes, 1960. PMID- 21643900 TI - Potential benefits and limitations of enriched environments and cognitive activity on age-related behavioural decline. AB - The main aim of this chapter is to review preclinical studies that have evaluated interventions which may aid in preventing or delaying age-related behavioural decline. Animal models of Environmental Enrichment (EE) are useful for evaluating the influence of cognitive, physical and social stimulation in mitigating cognitive decline at different ages. The EE paradigm has been proposed as a non invasive treatment for alleviating age-related memory impairment and neurodegenerative diseases. While in this complex environment, rodents can be stimulated at different levels (physical, social, cognitive and sensorial), although a synergism between all these components is likely to play an important role. We will summarize available data relating to EE as a potential therapeutic strategy that slows down or counteracts age-related cognitive and behavioural changes. EE also alters physiological responses and induces neurobiological changes such as stimulation of neurogenesis and neural plasticity. At the behavioural level, EE improves learning and memory tasks and reduces anxiety. Several variables seem to influence the behavioural and cognitive benefits induced by EE, including the age at which animals are first exposed to EE, total period during which animals are submitted to EE, gender, the cognitive task evaluated, the drug administered and individual factors. Cognitive and physical stimulation of animals in enriched experimental environments may lead to a better understanding of factors that promote the formation of cognitive reserve (CR) and a healthier life in humans. In the present chapter we review the potential benefits of EE in aged rodents and in animal models of Alzheimer Disease (AD). Results obtained in preclinical models of EE may be relevant to future research into mental and neurodegenerative diseases, stress, aging and development of enviromimetics. Finally, we outline the main limitations of EE studies (variability between laboratories, difficulty of separating the different components of EE, gender of experimental subjects, individual differences in the response to EE), evaluating the potential benefits of enriched environments and the neurobiological mechanisms that underlie them. We conclude that there are experimental data which demonstrate the cognitive benefits of rearing rodents in enriched environments and discuss their implication for clarifying which variables contribute to the formation of the CR. PMID- 21643901 TI - Estrogens and gonadal function in schizophrenia and related psychoses. AB - Recent research has increasingly pointed to the importance of estrogens and the hypothalamic-pituitary-gonadal axis in schizophrenia. Specifically, there is mounting evidence from clinical, epidemiological, and basic research that estradiol, the main component of estrogens, exerts protective effects in schizophrenia and related psychoses. Possible modes of action of this hormone in the brain have been suggested, and clinical intervention studies have reported the first positive results. Furthermore, there are an increasing number of reports on gonadal dysfunction and states of estrogen deficiency in women with schizophrenia. These findings could have important implications for clinicians and researchers alike. PMID- 21643902 TI - European Union Regulations. AB - The European Union (EU) has been a leader in the development of both guidance and regulations to ensure food safety throughout the member states. Because of the free movement of food commodities among the countries that belong to the European Union, there is a great need to assure high quality monitoring of both imported food and member state products. The procedures and methods required need to be practical, state-of-the art, and harmonised. The European Commission has developed a network of laboratories and scientific studies to meet this goal. This chapter describes the current Regulations, Directives and Decisions of the European Commission that protect the food supply throughout Europe. Because imported food needs to comply with the EU requirements, and the need to have common compliance throughout the member states, the developed system could be a worldwide template for monitoring the food supply. In addition, the integral role of chromatography hyphenated to mass spectrometry is described. PMID- 21643903 TI - China's food safety regulation and mass spectrometry. AB - Food safety is essential to people's health and people's livelihood. To ensure that food safety is an important current strategy of the governments, both regulation and standardization are important support for implementing this strategic initiative effectively. The status and prospects of China's food laws, regulations, and standards system are introduced. China now has established a complete law regime providing a sound foundation and good environment for keeping the health of people, maintaining the order of social economy and promoting the international trade of food. At the same time, it is undoubtedly important to strengthen standardization and improve the food safety standards system. In the administration of food safety, mass spectrometry is becoming more and more important and many analytical methods developed in China are based on its application. PMID- 21643904 TI - United States and Japanese food regulations. AB - The two other major importers and exporters of the world are the USA and Japan. Each of their regulations impact how the network of food supply and delivery are viewed. Their regulations mirror that of Europe but have their own origins and focal points. It is the intent of this chapter to provide an overview of food safety through the regulatory bodies of these two world influencers. This should provide a more complete picture of the current state of implementation for keeping the food supply safe for the people of the world. PMID- 21643905 TI - QuEChERS sample preparation approach for mass spectrometric analysis of pesticide residues in foods. AB - This chapter describes an easy, rapid, and low-cost sample preparation approach for the determination of pesticide residues in foods using gas and/or liquid chromatographic (GC and/or LC) analytical separation and mass spectrometric (MS) detection. The approach is known as QuEChERS, which stands for "quick, easy, cheap, effective, rugged, and safe." Originally, QuEChERS was a particular "method" for pesticide residue analysis, but it is very flexible and has evolved into an "approach," which has been used in many methods, and not just for pesticide residues. Two of the QuEChERS versions using buffering have been validated in interlaboratory trials for dozens of pesticides in several food matrices, and both have successfully met performance criteria to achieve "official" status from international standard organizations (AOAC Official Method 2007.01 and CEN Standard Method EN 15662). The main aspects of the QuEChERS approach consists of extraction of a well-homogenized sample by shaking with solvent (typically acetonitrile) in a centrifuge tube, salt-out partitioning of water with salts including magnesium sulfate (MgSO(4)), and cleanup using "dispersive solid-phase extraction" (dSPE), in which common matrix components are retained by sorbent(s) and the analytes remain in the extract. For widest analytical scope, concurrent analysis is done for hundreds of pesticides using GC MS(/MS) and LC-MS/MS. The aim of this chapter is to review the QuEChERS sample preparation methodology and provide a summary of up-to-date information with modification options depending on the application needs. PMID- 21643906 TI - Automated solid phase extraction. AB - An overview is given in this chapter of the main potential benefits of using automated Solid phase Extraction (SPE) in the preparation of food samples for LC MS analysis, both in terms of quality of results and in terms of performance and productivity. Automated SPE instrumentation is described and a range of application examples are given. The foods used in these applications range from non-fatty vegetables, to more complex vegetables, fish, prawn meat, and water, a vital raw product for the food and beverage industries. In most applications previously reported, the SPE technique was mainly used for sample cleanup prior to analysis. Additional examples are given here in which automated SPE is combined with analyte concentration, analyte derivatization, or addition of liquids such as internal standards to further improve limits of quantitation, selectivity, stability and quality of the analysis. PMID- 21643907 TI - Multiresidue pesticide analysis by capillary gas chromatography-mass spectrometry. AB - A multiresidue pesticide method using a modified QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) procedure and capillary gas chromatography-mass spectrometry (GC-MS) is described for the determination of 166 organochlorine, organophosphorus, and pyrethroid pesticides, metabolites, and isomers in spinach. The pesticides from spinach were extracted using acetonitrile saturated with magnesium sulfate and sodium chloride, followed by solid-phase dispersive cleanup using primary-secondary amine and graphitized carbon black sorbents and toluene. Analysis is performed using different GC-MS techniques emphasizing the benefits of non-targeted acquisition and targeted screening procedures. Non-targeted data acquisition of pesticides in the spinach was demonstrated using GC coupled to a single quadrupole mass spectrometery (GC-MS) in full scan mode or multidimensional GC-time-of-flight mass spectrometery (GC * GC-TOF/MS), along with deconvolution software and libraries. Targeted screening was achieved using GC-single quadrupole mass spectrometry in selective ion monitoring (GC-MS/SIM) mode or -tandem mass spectrometry (GC-MS/MS) in multiple reaction monitoring mode. The development of these techniques demonstrates the powerful use of GC-MS for the screening, identification, and quantitation of pesticide residues in foods. PMID- 21643908 TI - Targeted pesticide residue analysis using triple Quad LC-MS/MS. AB - The determination of pesticide residues by HPLC-MS/MS requires decisions on a multitude of analytical parameters. This includes the selection of eluents, columns and ion sources, but also the optimization of the tandem mass spectrometer for the selected target analytes. Another aspect is the use of the restricted acquisition time between two chromatographic data points. An appropriate selection of all these parameters as well as the measures to avoid interference by cross talks and wrong quantitative results by matrix effects is discussed in this chapter. PMID- 21643909 TI - LC/TOF-MS analysis of pesticides in fruits and vegetables: the emerging role of accurate mass in the unambiguous identification of pesticides in food. AB - The detection, identification, confirmation, and quantitation of pesticides in fruits and vegetables are typically performed from a list of suspect compounds or targets. However, there is mounting concern that pesticides not targeted are finding their way into the food supply. This chapter describes the use of LC with time-of-flight mass spectrometry (LC/TOF-MS) for the detection and identification of pesticides that are not targeted. The use of accurate mass measurement and its implication for the identification of non-targeted compounds are discussed. The need for unambiguous identification and requirements therein are evaluated in detail. PMID- 21643910 TI - Hormone analysis in food products. AB - The administration of growth-promoting compounds, to food producing animals is banned within the European Union. We developed several methods whereby the cleanup is based on LLE and SPE and detection based on GC-MSMS or LC-MSMS to identify and confirm the identity of different growth promoting agents in several food products. This chapter describes methods to isolate and identify these growth promoting agents. PMID- 21643911 TI - Analysis of multiple mycotoxins in food. AB - Mycotoxins are secondary metabolites of microscopic filamentous fungi. With regard to the widespread distribution of fungi in the environment, mycotoxins are considered to be one of the most important natural contaminants in foods and feeds. To protect consumers' health and reduce economic losses, surveillance and control of mycotoxins in food and feed has become a major objective for producers, regulatory authorities, and researchers worldwide. In this context, availability of reliable analytical methods applicable for this purpose is essential. Since the variety of chemical structures of mycotoxins makes impossible to use one single technique for their analysis, a vast number of analytical methods has been developed and validated. Both a large variability of food matrices and growing demands for a fast, cost-saving and accurate determination of multiple mycotoxins by a single method outline new challenges for analytical research. This strong effort is facilitated by technical developments in mass spectrometry allowing decreasing the influence of matrix effects in spite of omitting sample clean-up step. The current state-of-the-art together with future trends is presented in this chapter. Attention is focused mainly on instrumental method; advances in biosensors and other screening bioanalytical approaches enabling analysis of multiple mycotoxins are not discussed in detail. PMID- 21643912 TI - Multi mycotoxin analysis in food products using immunoaffinity extraction. AB - We developed a method for the simultaneous determination of deoxynivalenol, T-2 toxin, HT-2 toxin and zearalenone in wheat and biscuit by liquid chromatography/electrospray ionization/tandem mass spectrometry coupled with immunoaffinity extraction. This chapter describes a method to extract, clean-up, and quantitate these mycotoxins and the effect of the ion suppression of multifunctional column and IAC in the clean-up were compared. PMID- 21643913 TI - Multiresidue analysis of antibiotics in food of animal origin using liquid chromatography-mass spectrometry. AB - Antibiotics are the most important drugs administered in veterinary medicine. Their use in food-producing animals may result in antibiotic residues in edible tissues, which are monitored to protect human and animal health, support the enforcement of regulations, provide toxicological assessment data, and resolve international trade issues. This chapter provides basic characterization of the most important classes of antibiotics used in food-producing animals (aminoglycosides, amphenicols, beta-lactams, macrolides and lincosamides, nitrofurans, quinolones, sulfonamides, and tetracyclines), along with examples of practical liquid chromatographic-(tandem) mass spectrometric methods for analysis of their residues in food matrices of animal origin. The focus is on multiresidue methods that are favored by regulatory and other food testing laboratories for their ability to analyze residues of multiple compounds in a time- and cost effective way. PMID- 21643914 TI - The LC-MS/MS methods for the determination of specific antibiotics residues in food matrices. AB - This chapter describes the LC-MS/MS methods for the determination of antibiotics residues in food matrices. The types of antibiotics include beta-lactam antibiotics, sulfonamides, tetracyclines, fluoroquinolones, nitrofurans, and chloramphenicol (CAP). The food matrices are mainly from animal origin, such as animal tissues, fishes (marine products), eggs, milk, honey, and so on. The methods and procedures are covered, including three parts: (1) Liquid chromatographic conditions, (2) mass spectrometer conditions, including ionization source, analyzer, and acquisition, and (3) extraction and clean-up methods. In each case, the standard operating procedures (SOPs) for analysis are given with sensitivity, linearity, precision, and recovery. Some criteria of maximum residue limits (MRLs) from the legislation are listed. PMID- 21643915 TI - Identification of unknown migrants from food contact materials. AB - Materials that come into contact with foodstuffs can transfer components that may cause odour or taint problems or in the worse case cause the foodstuff to be unsafe to eat. The identities of some of these are easily predicted from the chemistry of known components but others are not. In this respect, it is important to be able to identify and quantify these chemicals. This chapter describes the need for methods of identification of unknown chemicals that may migrate. Mass spectrometric analytical methods are described, including headspace gas chromatography with mass spectrometry (HS-GC-MS), liquid injection gas chromatography with MS, and liquid chromatography with time-of-flight MS (LC-TOF MS). PMID- 21643916 TI - Halogenated persistent organic pollutants and polycyclic aromatic hydrocarbons in food. AB - During recent years, mass spectrometry (MS) and hyphenated chromatographic instrumentation and techniques have been a subject of dramatic developments, resulting in the introduction of various useful tools for the analysis of halogenated persistent organic pollutants (POPs) and polycyclic aromatic hydrocarbons (PAHs) in food and environmental matrices. This chapter describes state-of-the-art in the field of MS as a primary detection tool for the halogenated POPs and PAHs previously separated using either gas chromatography (GC) or liquid chromatography (LC). Since sample preparation practice plays a crucial role for obtaining optimal performance characteristics of a particular analytical method, a brief overview of sample extraction and clean-up procedures in the POPs/PAHs analysis is also briefly outlined. PMID- 21643917 TI - Salivary cortisol and alpha-amylase reactivity to taekwondo competition in children. AB - The aim of this study was to evaluate the effects of an official taekwondo competition (three 1-min rounds with a 1-min recovery in-between) on heart rate (HR), salivary alpha-amylase (sAA), and salivary-free cortisol (sC) in children. Parental consent was obtained for 12 young (10.4 +/- 0.2 years) male taekwondo athletes. Saliva sample were collected 15 min before and 1 min after an official taekwondo competition, and at 30, 60, and 90 min of the recovery period. To evaluate the exercise intensity during the competition, HR was measured and expressed as a percentage of individuals HR(peak). Athletes spent 78% of the time working at HR > 90% HR(max), with significant increases from round 1 to round 2 and 3. Peak sAA observed at the end of the match (169.6 +/- 47.0 U/mL) was different (P = 0.0001) from the other samplings (pre-competition 55.0 +/- 14.0 U/mL, 30-min recovery 80.4 +/- 17.7 U/mL, 60-min recovery 50.5 +/- 7.6 U/ml; 90 min recovery 53.2 +/- 9.6 U/mL). Peak sC values observed at 30-min recovery (17.9 +/- 3.5 nmol/L) were different (P < 0.0001) from pre-competition (5.6 +/- 0.9 nmol/L), post-competition (9.0 +/- 2.0 nmol/L), 60-min recovery (10.3 +/- 2.6 nmol/L) and 90-min recovery (4.2 +/- 0.8 nmol/L) values. These findings confirm that taekwondo competitions pose a high stress on young athletes. The different sAA and sC reactions in response to the physical stressor mirror the faster reactivity of the sympathetic-adrenomedullary system relatively to the hypothalamic-pituitary-adrenocortical system, respectively. This experimental paradigm might represent a useful model for further research on the effects of various stressors (i.e., training and competition) in taekwondo athletes. PMID- 21643918 TI - The effects of a 30-min run on the mechanics of the human Achilles tendon. AB - Tendinous structures often exhibit reduced stiffness following repeated loading via static muscular contractions. The purpose of this study was to determine if human Achilles tendon (AT) stiffness is affected by the repeated loading experienced during running and if this affects normal muscle-tendon interaction. Twelve male participants (mean +/- SD: age 27 +/- 5 years, height 1.79 +/- 0.06 m, mass 78.6 +/- 8.4 kg) completed a 30 min run at 12 kmph on a treadmill. AT properties were determined before and after the run during a series of one-legged hops. During hopping and running, AT length data were acquired from a combination of ultrasound imaging (50 Hz) and kinematic data (200 Hz). AT force was estimated from inverse dynamics during hopping and AT stiffness was computed from plots of AT force and length. AT stiffness was not significantly different post run (pre 163 +/- 41 N mm(-1), post 147 +/- 52 N mm(-1), P > 0.05) and peak AT strain during the stance phase of running (calculated relative to AT length during standing) was similar at different time points during the run (3.5 +/- 1.8% at 1 min, 3.2 +/- 1.8% at 15 min and 3.8 +/- 2% at 30 min). It was concluded that the loading experienced during a single bout of running does not affect the stiffness of the AT and that the properties of the AT are stable during locomotion. This may have implications for muscle fascicle behaviour and Achilles tendon injury mechanisms. PMID- 21643919 TI - Autoregulation in the ocular and cerebral arteries during the cold pressor test and handgrip exercise. AB - The aim of this study was to determine whether autoregulation exerts similar effects in the ocular and cerebral vessels, which are both branches of the internal carotid artery. Ocular blood flow velocities, cerebral blood flow velocity and blood pressure were measured in 11 subjects during a 2-min resting period, static handgrip exercise (HG) and a cold pressor test (CPT). Blood velocity data for the superior and inferior temporal retinal arterioles (STRA and ITRA, respectively) and the retinal and choroidal vasculature (RCV) were obtained for 4 s during the measurement using laser speckle flowmetry. Mean blood flow velocity in the middle cerebral artery (MCAVmean) was measured by transcranial Doppler ultrasound. The conductance index (CI) of each vessel was calculated by dividing blood flow by mean arterial pressure. Blood flow velocity in the RCV increased by 19 +/- 9% from resting baseline level during the CPT (P < 0.05), while blood flow in the STRA, ITRA and MCAVmean did not. The CI of the MCA decreased. The RCV blood flow velocity, ITRA blood flow and MCAVmean increased by 8 +/- 1, 9 +/- 3 and 11 +/- 4%, respectively, during the HG (P < 0.05). Conversely, STRA blood flow remained unchanged. The HG did not significantly change the CI in any of the vessels measured. These findings suggest that cerebral blood flow velocity was maintained during the CPT, but autoregulation does not work well in the RCV during the CPT and HG. PMID- 21643920 TI - Neural adaptations to electrical stimulation strength training. AB - This review provides evidence for the hypothesis that electrostimulation strength training (EST) increases the force of a maximal voluntary contraction (MVC) through neural adaptations in healthy skeletal muscle. Although electrical stimulation and voluntary effort activate muscle differently, there is substantial evidence to suggest that EST modifies the excitability of specific neural paths and such adaptations contribute to the increases in MVC force. Similar to strength training with voluntary contractions, EST increases MVC force after only a few sessions with some changes in muscle biochemistry but without overt muscle hypertrophy. There is some mixed evidence for spinal neural adaptations in the form of an increase in the amplitude of the interpolated twitch and in the amplitude of the volitional wave, with less evidence for changes in spinal excitability. Cross-sectional and exercise studies also suggest that the barrage of sensory and nociceptive inputs acts at the cortical level and can modify the motor cortical output and interhemispheric paths. The data suggest that neural adaptations mediate initial increases in MVC force after short-term EST. PMID- 21643921 TI - Cognitive reserve and its implications for rehabilitation and Alzheimer's disease. AB - According to the Cognitive reserve hypothesis, several factors related to mental engagement, such as level of education, type of occupation, leisure activities and social network, appear to affect the risk of developing clinical dementia. The present article provides an overview of the studies that have investigated the effects of mental engagement and cognitive stimulation specifically on dementia of the Alzheimer's type (AD). Mental training and cognitive stimulation interventions in AD have been shown to be useful in increasing patients' ability in performing activities of daily living (ADL), allowing them to maintain relative independence. Since cognitive engagement and stimulation are known to modify the brain processes to perform tasks, by recruiting alternative and more efficient networks, this review is especially focused on cognitive rehabilitation in AD patients, which has been shown to improve their global functioning and cognition. This perspective stresses the idea that cognitive reserve is not a fixed factor, but can be continuously modified by life experiences, even when the brain is already affected by neuropathology. PMID- 21643922 TI - Spontaneous healing in complete ACL ruptures: a clinical and MRI study. AB - BACKGROUND: Most authors believe the ACL does not spontaneously heal after a complete rupture. Although several studies have reported spontaneous healing of torn ACLs, it is difficult to determine its healing potential and whether patients will be able to return to sports activities. QUESTIONS/PURPOSES: We therefore asked whether (1) a complete ACL rupture in patients can spontaneously heal without the use of a specific rehabilitation program or bracing and (2) patients are able to return to their athletic activity after spontaneous ACL healing. PATIENTS AND METHODS: We retrospectively reviewed 14 patients with acute ACL injury established by physical examination and MRI (proximal third in eight patients and the midligament in six). Average age at injury was 31 years (range, 23-41 years). All patients were athletically active before injury. Surgery was indicated in all patients but for various reasons postponed. We obtained International Knee Documentation Committee scores, Lysholm-Gillquist scores, and MRI. The minimum followup was 25 months (mean, 30 months; range, 25-36 months). RESULTS: At last followup, the mean Lysholm-Gillquist score was 97. According to the International Knee Documentation Committee evaluation, 10 knees were normal and four nearly normal. All knees regained end point with a negative pivot shift test; MRI at followup showed an end-to-end continuous ACL with homogeneous signal. All patients returned to their former activity level. However, after the study period, two patients had a rerupture of the ACL (2.5 years after the first lesion). CONCLUSIONS: Our observations indicate an acutely injured ACL may eventually spontaneously heal without using an extension brace, allowing return to athletic activity. LEVEL OF EVIDENCE: Level IV, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence. PMID- 21643923 TI - Cost analysis of a falls-prevention program in an orthopaedic setting. AB - BACKGROUND: Falls by orthopaedic patients may lead to negative outcomes such as injury, prolonged hospitalization, delayed rehabilitation, and increased costs. QUESTIONS/PURPOSES: We examined the impact of a multidisciplinary Falls prevention Program (FPP) on the incidence of inpatient falls and fall-related injuries in an orthopaedic hospital during a 6-year period. METHODS: Patient data and fall incident report data were reviewed to identify risk factors associated with falls and fall-related injuries. A cost analysis was performed to calculate costs incurred as a result of falls. RESULTS: A total of 415 falls occurred during a 5-year period preintervention. The fall rate preintervention was significantly higher than the fall rate postintervention (3.49 versus 2.68 per 1000 bed days). Eighty-five falls occurred in the 12 months preintervention. A total of 15.29% (13 of 85) of falls resulted in minor injuries, and 9.42% (eight of 85) resulted in major injuries. The total cost incurred during this period as a result of falls was $117,754.12. Of this, 95.5% resulted from patients who sustained a hip fracture (n = 4). The total cost of implementing the FPP was $15,694.46. In the 12 months postintervention, 52 falls occurred. Twenty-five percent (13 of 52) of falls resulted in minor injuries, and 5.76% (two of 52) resulted in major injuries (no hip fractures). The total costs accrued during this period as a result of falls was $811.70. CONCLUSIONS: After implementation of a FPP, there were significant decreases in fall incidence, fall-related morbidity, and consequent costs. LEVEL OF EVIDENCE: Level III, economic and decision analyses. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 21643924 TI - Does open reduction of the developmental dislocated hip increase the risk of osteonecrosis? AB - BACKGROUND: Osteonecrosis (ON) of the femoral head is one of the main complications associated with treatment of developmental dysplasia of the hips (DDH). The reported rates of ON vary widely between 6% and 48%, suggesting varying factors in these studies influence the rate. Several studies suggest open reduction combined with femoral shortening provides protection against ON. However, it is unclear whether confounders such as failed Pavlik harness treatment, preliminary traction, closed versus open reduction, and redislocation influence the rate of ON. QUESTIONS/PURPOSE: We therefore asked whether open reduction with concomitant osteotomies without femoral shortening, redislocation, and secondary surgical procedures for residual acetabular dysplasia influenced the rate of ON. METHODS: We retrospectively reviewed 64 children (78 hips) hospitalized with developmental dislocation of the hip between January 1998 and February 2007. Patients younger than 12 months were treated with closed or open reduction. Open reduction combined with concomitant pelvic and femoral osteotomies was performed in patients past walking age. ON was diagnosed from radiographs obtained at last followup. We used logistic regression analysis to identify predictors for the development of ON. The minimum followup was 3.2 years (mean, 6.8 years; range, 3.2-11.5 years). RESULTS: The overall rate of ON was 40%. Patients who underwent open reduction combined with concomitant osteotomies, experienced redislocation, or required secondary reconstructive procedures after initial reduction were at higher risk for having ON develop. CONCLUSIONS: We advocate early reduction of the dislocated hip in the first year of life to avoid the need for concomitant osteotomies combined with open reduction. PMID- 21643925 TI - The use of an algorithm for classifying acetabular fractures: a role for resident education? AB - BACKGROUND: The Letournel and Judet classification system is commonly used for classifying acetabular fractures. However, for orthopaedic surgeons with less experience with these fractures, correct classification can be more difficult. A stepwise approach has been suggested to enhance the inexperienced observer's ability to properly classify acetabular fractures, but it is unclear whether this actually improves one's ability. QUESTIONS/PURPOSES: We asked (1) whether the use of a step-by-step algorithm improves residents' ability to classify acetabular fractures, (2) whether resident experience influenced ability to correctly classify acetabular fractures, and (3) which acetabular fractures were the most difficult to classify? METHODS: Forty-six residents reviewed 15 sets of plain radiographs of 10 acetabular fracture patterns. Residents used the Letournel and Judet classification with only a diagram for reference. Three weeks later they were asked to classify the fractures a second time with the use of the algorithm. We then compared the number of correct responses from the two sessions and determined whether resident experience and use of the algorithm influenced correct classification. RESULTS: We found an improvement in the number of correctly classified fractures between the first (348/690 [50%]) and second (409/690 [59%]) sessions. Thirty-two of 46 participants improved their score with the use of the algorithm. There was a tendency for participants with more residency training to correctly classify the fractures. CONCLUSIONS: The algorithm provided modest improvement to the residents' ability to classify acetabular fractures. This or other such algorithms could provide residents with a basic tool to better evaluate standard radiographs and classify acetabular fractures. PMID- 21643926 TI - Wellness as fairness. AB - I argue that distinct conditions of justice lead to diverse wellness outcomes through a series of psychosocial processes. Optimal conditions of justice, suboptimal conditions of justice, vulnerable conditions of injustice, and persisting conditions of injustice lead to thriving, coping, confronting, and suffering, respectively. The processes that mediate between optimal conditions of justice and thriving include the promotion of responsive conditions, the prevention of threats, individual pursuit, and avoidance of comparisons. The mechanisms that mediate between suboptimal conditions of justice and coping include resilience, adaptation, compensation, and downward comparisons. Critical experiences, critical consciousness, critical action, and righteous comparisons mediate between vulnerable conditions of injustice and confrontation with the system. Oppression, internalization, helplessness, and upward comparisons mediate between persisting conditions of injustice and suffering. These psychosocial processes operate within and across personal, interpersonal, organizational and community contexts. Different types of justice are hypothesized to influence well being within each context. Intrapersonal injustice operates at the personal level, whereas distributive, procedural, relational, and developmental justice impact interpersonal well-being. At the organizational level, distributive, procedural, relational and informational justice influence well-being. Finally, at the community level, distributive, procedural, retributive, and cultural justice support community wellness. Data from a variety of sources support the suggested connections between justice and well-being. PMID- 21643927 TI - An uncommon side-effect of linezolid. PMID- 21643928 TI - The management of chronic kidney disease and end-stage renal disease in Nigeria. PMID- 21643929 TI - The effect of marital status on stage and survival of prostate cancer patients treated with radical prostatectomy: a population-based study. AB - OBJECTIVE: The detrimental effect of unmarried marital status on stage and survival has been confirmed in several malignancies. We set to test whether this applied to patients diagnosed with prostate cancer (PCa) treated with radical prostatectomy (RP). METHODS: We identified 163,697 non-metastatic PCa patients treated with RP, within 17 Surveillance, Epidemiology, and End Results registries. Logistic regression analyses focused on the rate of locally advanced stage (pT3-4/pN1) at RP. Cox regression analyses tested the relationship between marital status and cancer-specific (CSM), as well as all-cause mortality (ACM). RESULTS: Respectively, 9.1 and 7.8% of individuals were separated/divorced/widowed (SDW) and never married. SDW men had more advanced stage at surgery (odds ratio: 1.1; p < 0.001), higher CSM and ACM (both hazard ratio [HR]: 1.3; p < 0.001) than married men. Similarly, never married marital status portended to a higher ACM rate (HR:1.2, p = 0.001). These findings were consistent when analyses were stratified according to organ confined vs. locally advanced stages. CONCLUSIONS: Being SDW significantly increased the risk of more advanced stage at RP. Following surgery, SDW men portended to a higher CSM and ACM rate than married men. Consequently, these individuals may benefit from a more focused health care throughout the natural history of their disease. PMID- 21643930 TI - Telomere length in peripheral blood and breast cancer risk in a prospective case cohort analysis: results from the Sister Study. AB - OBJECTIVE: Telomeres are required for maintaining genomic integrity and may play a role in carcinogenesis. Some, but not all, epidemiologic studies have found that short telomeres in leukocytes are associated with an increased risk of breast cancer. To further elucidate this potential association, we examined telomere length in relation to breast cancer risk in prospectively collected blood samples from the Sister Study, a cohort of women aged 35-74 years who have a sister with breast cancer. METHODS: We performed a case-cohort analysis comparing incident breast cancer cases (n = 342) with a subcohort (n = 735), randomly selected from 29,026 participants, enrolled by June 1, 2007. Relative telomere length in peripheral blood cells was estimated using a single-tube monochrome multiplex quantitative PCR assay. RESULTS: No association was observed between telomere length and breast cancer risk. Compared with the longest quartile, hazard ratios (HR) associated with the second, third, and the shortest quartile were 0.91 [95% confidence interval (95% CI): 0.62-1.34], 1.11 (95% CI: 0.77-1.60), and 0.93 (95% CI: 0.64-1.35), respectively. Subgroup analyses by menopausal status, invasiveness, or estrogen receptor status of breast cancer did not reveal evidence of association between telomere length in blood cells and subsequent breast cancer risk. CONCLUSIONS: This prospective investigation does not support telomere length in blood cells as a biomarker for breast cancer risk. PMID- 21643931 TI - Predictors of waterpipe smoking among secondary school adolescents in Al Hassa, Saudi Arabia. AB - BACKGROUND: There has been a global upsurge of waterpipe (WP) smoking over the past several years. This can be attributed at least partly to various factors like lack of knowledge regarding its health effects, social acceptability, and intensive preventive programs focusing selectively on cigarettes smoking. PURPOSE: This study aimed to determine the prevalence and psychosocial determinants of WP smoking among secondary school adolescents in Al Hassa, Saudi Arabia, and also to assess their attitudes toward WPs smoking and knowledge regarding the related health effects. METHODS: Cross-sectional study included 1,652 students of both genders selected by multistage proportionate sampling method. Data collection was carried out using a self-administered anonymous Arabic version of the Global Youth Tobacco Survey, modified with items dedicated to WP smoking. Items to assess participants' knowledge about the health-related hazards and attitude towards WP were added. Patient Health Questionnaire was used to screen for the presence of anxiety and depressive disorders. RESULTS: Of the included adolescents, 358 (21.7%) were current smokers; cigarettes (46.1%), both cigarettes and WP (16.5%), and only WP (37.4%). Current WP users accounted for 193; 96.4% were males and 62.7% were >=18 years of age. Outing, boredom, meeting friends, and family gatherings were the most frequently stated motives. WP users demonstrated a higher incidence of depressive and generalized anxiety disorders. Common assumptions regarding WP smoking included: WP smoking is less harmful than cigarettes and with no addictive property as stated by 47.8% and 65.9% of adolescents, respectively, harmful substances are purified through water filtration as believed by 59.2%, and it is more socially acceptable than cigarettes as agreed by 54.1%. Hierarchical regression analysis showed that male gender and increasing age of adolescents, WP smoking among close family and friends, and socializing motives were significant predictors for the current WP smoking state. CONCLUSION: Social acceptability, poor knowledge of WP health related hazards, and socio-demographics are favoring the current increasing trend of WP use among adolescents in Al Hassa, Saudi Arabia. PMID- 21643932 TI - Cytogenetic analysis of a malignant triton tumour by comparative genomic hybridization (CGH) and review of the literature. AB - Malignant triton tumour (MTT) is a rare, highly malignant neoplasm, characterized by a mixture of cells with nerve sheath and skeletal muscle differentiation. Cytogenetic analyses of this neoplasm are rare to date and none comparative genomic hybridisation (CGH) analysis has been published. In the present study we report about the genomic imbalances of a MMT analysed by CGH, in a 39-year-old male patient without neurofibromatosis. We observed the amplifications at chromosomal location 1p, 6p, 16p, 16q, 17p, 17q, 19p, 19q, 20p, and 22q. Comparing our results with those of previous studies, we found evidence for recurrent genomic aberrations at the chromosomes 1, 16, 17, 19, and 22 suggesting the involvement of several oncogenes in the genesis of MTT. PMID- 21643933 TI - Exercise-induced laryngeal obstruction: natural history and effect of surgical treatment. AB - The current follow-up study concerning the supraglottic type of exercise-induced laryngeal obstruction (EILO) was performed to reveal the natural history of supraglottic EILO and compare the symptoms, as well as the laryngeal function in conservatively versus surgically treated patients. A questionnaire-based survey was conducted 2-5 years after EILO was diagnosed by a continuous laryngoscopy exercise (CLE) test in 94 patients with a predominantly supraglottic obstruction. Seventy-one patients had been treated conservatively and 23 with laser supraglottoplasty. The questionnaire response rate was 70 and 100% in conservatively treated (CT) and surgically treated (ST) patients, respectively. A second CLE test was performed in 14 CT and 19 ST patients. A visual analogue scale on symptom severity indicated improvements in both the groups, i.e. mean values (+/- standard deviations) declined from 73 (20) to 53 (26) (P < 0.001) in the CT group and from 87 (26) to 25 (27) (P < 0.001) in the ST group. At follow up, ST patients reported lower scores regarding current level of complaints, and higher ability to perform exercise, as well as to push themselves physically, all compared to CT patients (P < 0.001). CLE scores were normalized in 3 of 14 (21%) CT and 16 of 19 (84%) ST patients (Z = -3.6; P < 0.001). In conclusion, symptoms of EILO diagnosed in adolescents generally decreased during 2-5 years follow-up period but even more after the surgical treatment. Patients with supraglottic EILO may benefit from supraglottoplasty both as to laryngeal function and symptom relief. PMID- 21643934 TI - The effects of repetitious topical use of mitomycin C on antrostomy patency in maxillary antrostomy created rabbit model. AB - The aim of our study was to investigate the effect of the topical use of mitomycin C (MMC) intraoperatively in single dose and intra-postoperatively in two doses on the narrowing of antrostomy in maxillary rabbit sinus antrostomies created experimentally. And also to determine the local and systemic side effects of topical MMC. With this objective, 0.6 mg/ml MMC was used to the first group at single dose and to the second group intraoperatively and on third day postoperatively in two doses topically for 5 min. After 8 weeks, although the mean area of antrostomy was larger than that in the control side in the first group, which received single dose MMC, the difference was not statistically significant (p = 0.287). The second group received two doses, and the antrostomy areas were found to be significantly larger than the controls (p = 0.05). Overall, the sides that received MMC were significantly larger (p = 0.029). From the point of histopathological examination of the tissue, it was seen that two dose MMC increased the edema indicating inflammation and antrostomy resolved with normal respiratory tract epithelium. It was shown by measuring the blood values that nephrotoxic and myelosupressant effect of MMC occurring in systemic use did not occur with single or double dose topical use. Our results demonstrate that even if the number of cases was low, two doses of topical MMC usage prevent the narrowing of antrostomy while single dose MMC does not. And two-dose topical MMC usage does not have local and systemic side effects. PMID- 21643935 TI - Functional outcome in patients with advanced head and neck cancer: surgery and reconstruction with free flaps versus primary radiochemotherapy. AB - The objective was to compare functional outcome in patients with advanced head and neck cancer (HNC) treated with (a) surgical resection and reconstruction with microvascular free flaps (MVFF) followed by radiochemotherapy versus (b) primary radiochemotherapy (RCT) on the basis of the International Classification of Functioning, Disability and Health (ICF) from WHO. This was a cross-sectional, multi-institutional study. The outcome measures included ICF Core Sets for HNC, the EORTC-QLQ, modules c30 + hn35 and the University of Washington-Quality of life Questionnaire (UW-QOL). Analyses included descriptive statistics, ranking exercises, and regression analyses in a cumulative logit model; 27 patients were treated with MVFF and 22 with RCT. Global Quality of life scores suggested a slightly better functional outcome for the surgical approach. The majority of ICF categories (81/93, 87%) did not show a difference in functional outcome between the two treatment approaches. In the remaining 12 ICF categories, n = 3 body structures were more affected in the MVFF group, while n = 3 body functions, and n = 6 activities/participations were more problematic in the RCT group. This included oral swallowing and weight maintenance functions as well as social relationships, acquiring a job, and economic self-sufficiency. In addition, nine contextual environmental factors were more relevant to the RCT group. Both treatment approaches seemed appropriate to advanced HNC from the perspective of functional outcome. The influence of treatment modalities on the social and economic lives of cancer survivors needs to be explored further. In order to guide rehabilitation according to patients' needs, the ICF offers a multidimensional view comprising body structures, body functions, and activities and participation in life. PMID- 21643936 TI - The association between serious psychological distress and emergency department utilization among young adults in the USA. AB - PURPOSE: Psychological problems could lead to several adverse health outcomes and were strongly correlated with cigarette smoking and alcohol consumption. In addition, patients treated in EDs were vulnerable to psychological problems. We therefore examined the population-level association between serious psychological distress (SPD) and emergency department (ED) use among young adults in the USA. We also studied the additive effects of SPD, cigarette smoking, and alcohol consumption on the ED presentation. METHODS: The study sample contains 16,873 individuals, using data from the National Health Interview Survey, from 2004 to 2006. Bivariate analyses with chi-square tests and logistic regression analyses are performed. RESULTS: Young adults having SPD were 2.05 times more likely to go to an ED. People having SPD and being a current smoker were 2.52 times more likely to use services in an ED. However, people having SPD and being a heavy drinker did not have a significantly elevated risk of ED use. CONCLUSION: An association between SPD and ED use among US young adults is established in this study. Attempts to decrease excess ED use and the development of strategies to improve mental health among young adults are needed to improve patient health and reduce the health-care burden of high costs and deteriorating ED care quality. PMID- 21643937 TI - Cell-specific detection of microRNA expression during cardiomyogenesis by combined in situ hybridization and immunohistochemistry. AB - MicroRNAs (miRNAs) regulate gene expression by mediating translational repression or mRNA degradation of their targets, and several miRNAs control developmental decisions through embryogenesis. In the developing heart, miRNA targets comprise key players mediating cardiac lineage determination. However, although several miRNAs have been identified as differentially regulated during cardiac development and disease, their distinct cell-specific localization remains largely undetermined, likely owing to a lack of adequate methods. We therefore report the development of a markedly improved approach combining fluorescence based miRNA-in situ hybridization (miRNA-ISH) with immunohistochemistry (IHC). We have applied this protocol to differentiating embryoid bodies (EBs) as well as embryonic and adult mouse hearts, to detect miRNAs that were upregulated during EB cardiomyogenesis, as determined by array-based miRNA expression profiling. In this manner, we found specific co-localization of miR-1 to myosin positive cells (cardiomyocytes) of EBs, developing and mature hearts. In contrast, miR-125b and 199a did not localize to cardiomyocytes, as previously suggested for miR-199a, but were rather expressed in connective tissue cells of the heart. More specifically, by co-staining with alpha-smooth muscle actin (alpha-SMA) and collagen-I, we found that miR-125b and -199a localize to perivascular alpha-SMA( ) stromal cells. Our approach thus proved valid for determining cell-specific localization of miRNAs, and the findings we present highlight the importance of determining exact cell-specific localization of miRNAs by sequential miRNA-ISH and IHC in studies aiming at understanding the role of miRNAs and their targets. This approach will hopefully aid in identifying relevant miRNA targets of both the heart and other organs. PMID- 21643938 TI - Radiologic findings of osteoarticular infection in paracoccidioidomycosis. AB - OBJECTIVE: To report the radiological abnormalities of osteoarticular involvement in paracoccidioidomycosis (PCM). MATERIALS AND METHODS: After institutional board approval, the medical records and conventional radiology findings of 19 patients with osseous PCM were retrospectively reviewed. Number, distribution, and lesion characteristics were evaluated in consensus by two experienced musculoskeletal radiologists. RESULTS: The mean age of patients was 16.1 years (range 4-49 years), 11 male and eight female. MSK involvement was the only or the primary presentation of the disease in eight of 19 patients (42.1%). In total, 51 focal bone lesions were detected, being 41 in long bones. In long bones lesions, 19 of 41 (46.4%) were metaphyseal, 12 of 41 (29.3%) meta-epiphyseal, and 12 of 41 (29.3%) diaphyseal. The most common presentation was a geographic osteolytic bone lesion (62.7%), without marginal sclerosis (82.4%) and without periosteal reaction (90.2%). Articular involvement was present in six of 19 patients (31.6%), being two cases of primary arthritis. CONCLUSIONS: All encountered bone lesions were osteolytic. Metaphyseal or meta-epiphyseal osteomyelitis of a long bone was the most prevalent osteoarticular manifestation of paracoccidioidomycosis. PCM osteoarticular involvement could be solitary or multifocal, occurs almost exclusively in the acute/subacute clinical form, and it is more common in children and in juvenile patients. Axial skeleton involvement, arthritis, or a disseminated osseous pattern of infection may occasionally occur in this fungal disease. PMID- 21643939 TI - Clinical usefulness of diffusion-weighted MR imaging for detection of pancreatic cancer: comparison with enhanced multidetector-row CT. AB - PURPOSE: The aim of this study was to compare diffusion-weighted magnetic resonance imaging (DWI) and multidetector-row computed tomography (MDCT) for detection of primary pancreatic cancer by reviewing images of patients at high risk for pancreatic cancer with main pancreatic duct (MPD) dilatation shown by magnetic resonance cholangiopancreatography (MRCP). METHODS: From October 2007 to September 2009, 83 patients who had undergone both DWI and MDCT with MPD dilatation were identified and were reviewed by four readers (2 radiologists and 2 gastroenterology fellows). Diagnostic performance in pancreatic cancer detection was evaluated with 95% confidence intervals. Statistically significant differences in the detection of pancreatic cancer between DWI and MDCT were compared by receiver operating characteristics and the confidence of the diagnosis by the paired t test. RESULTS: Thirty-two of 83 patients were diagnosed with primary pancreatic cancer by histological evaluation of 15 surgical and 2 endoscopic ultrasound-guided fine needle aspiration samples, and by the clinical course for 15 lesions. Overall average accuracies of pancreatic cancer detection by the four readers were 84% with DWI and 86% with MDCT. CONCLUSION: Performance of DWI and MDCT was equivocal for detection of pancreatic cancer in a high-risk population with MPD dilatation. The combination of MRCP and DWI for detection of pancreatic cancer allowed identification of a high-risk population and tumor detection with a single imaging modality with no need for contrast medium. PMID- 21643940 TI - Late Dysphagia and dyspnea as complications of esophagogastroduodenoscopy in delayed pressure urticaria: case report. AB - Delayed pressure urticaria (DPU) is characterized by swelling in the area of sustained pressure on the skin. The reported case was a potentially life threatening complication due to mucosal edema following esophagogastroduodenoscopy (EGD). A 37-year-old man, suffering from severe DPU and chronic spontaneous urticaria, had undergone EGD due to dyspeptic symptoms. A few hours after the EGD procedure, the patient showed both dysphagia and dyspnea. A physical examination indicated massive tongue base and pharynx edema. We suggest that these symptoms were most likely due to the pressure exerted by EGD. No other apparent origins such as angioedema or late-phase allergic reaction to drugs were identified. One should be aware of the increased risk of developing airway and gastrointestinal obstruction during medical procedures associated with compression, such as EGD or endotracheal intubation, in DPU patients. PMID- 21643941 TI - Impact of microvascular obstruction and infarct size on left ventricular remodeling in reperfused myocardial infarction: a contrast-enhanced cardiac magnetic resonance imaging study. AB - Infarct size (IS) and microvascular obstruction (MO) following ST-elevation myocardial infarction (STEMI) reperfusion may affect left ventricular (LV) remodeling. We evaluated the impact of extent and transmurality of IS and MO in LV remodeling using contrast-enhanced cardiac magnetic resonance imaging (MRI). Thirty-six consecutive patients presenting with a first STEMI and undergoing contrast-enhanced cardiac MRI within 5 days of successful primary percutaneous coronary intervention (PPCI) were enrolled. Gadolinium-enhanced MRI at first passage and in delayed imaging was performed to assess MO and IS. LV remodeling was evaluated by echocardiography at 6-month-follow-up and defined as a percent increase in the LV end-diastolic volume >20%. Thirteen patients (36%) developed LV remodeling. IS and MO extent score was associated with LV remodeling (OR 1.5, 95% CI 1.02-2.38, P = 0.04, and OR 3.1, 95% CI 1.45-6.64, P = 0.003, respectively), along with IS and MO trasmurality (OR 1.4, 95% CI 1.007-2.12, P = 0.046, and OR 3.1, 95% CI 1.24-7.89, P = 0.016, respectively). Importantly, IS and MO extent score combination gave an OR of 3.4 (95% CI 1.4-7.9, P = 0.004) and the combination of IS and MO transmurality increased the OR to 4.8 (95% CI 1.5 15.2, P = 0.007). Finally, when combining simultaneously IS and MO extent score and transmurality the OR reached 5.3 (95% CI 3.34-18.2, P = 0.0008). The evaluation of both IS and MO extent and transmurality by MRI is of prognostic utility in patients undergoing PPCI. Importantly, IS and MO transmurality significantly increases the risk of adverse remodeling and should be routinely assessed in post-STEMI patients. PMID- 21643942 TI - Progression of coronary artery calcium in men affected by human immunodeficiency virus infection. AB - Cardiovascular risk is increased in HIV infected patients. We assessed progression of coronary artery calcium (CAC) in patients with HIV infection to identify factors that may help explain progression of atherosclerosis. Prospective, observational study of 132 HIV-infected men receiving chronic antiretroviral therapy (ART); we measured traditional atherosclerosis risk factors and assessed progression of CAC on sequential 64-slice CT scans at an average interval of 11 months (range 6-36). CAC score progression was defined as absolute and percentage change from baseline. During follow-up 45 patients (34%) showed absolute progression of CAC and 34 of them showed >15% yearly progression, a threshold previously associated with a high risk of myocardial infarction. Age, LDL cholesterol, visceral abdominal fat and current T-helper (CD4+) cell count were significantly associated with absolute CAC progression. Progression of subclinical atherosclerosis in HIV patients is associated with traditional coronary risk factors as well as HIV related factors such as the CD4+ cell count. Therefore, immunologic perturbations secondary to HIV infection may contribute to atherosclerosis progression. PMID- 21643943 TI - Insufficient protection by Neisseria meningitidis vaccination alone during eculizumab therapy. PMID- 21643944 TI - Vitamin D deficiency is associated with short stature and may influence blood pressure control in paediatric renal transplant recipients. AB - Vitamin D deficiency is common in adult renal transplant recipients, but data in children are scarce. Vitamin D is shown to have multiple effects on the cardiovascular system, renal function, and maintenance of bone health. We hypothesized that 25(OH)D deficiency is common in pediatric renal transplant recipients, and may be associated with hyperparathyroidism, short stature, renal function, and blood pressure control. We recruited 106 children during the winter/spring season who had a functioning renal transplant for at least 3 months. Twenty-five hydroxyvitamin D [25(OH)D] and 1,25-dihydroxyvitamin D [1,25(OH)(2)D] were measured and correlated with clinical and biochemical parameters. Of the renal transplant patients, 38% were 25(OH)D deficient, 54% had insufficient levels, and only 8% had adequate 25(OH)D levels. Despite alfacalcidol supplementation in 59 (56%) patients, parathyroid hormone was increased in 58 (55%) and showed an inverse correlation with 25(OH)D (p = 0.0003, r = 0.61) but not with 1,25(OH)(2)D levels. Height standard deviation score (SDS) correlated with 25(OH)D (p = 0.007, r = 0.42) and time post transplantation (p = 0.02, r = 0.23); both were significant and independent predictors of height SDS. 25(OH)D inversely correlated with systolic BP SDS (p = 0.02, r =-0.26); this association was lost on multiple regression analysis, but 25(OH)D was the only modifiable risk factor for hypertension. There was no correlation with estimated GFR or proteinuria. In conclusion, 25(OH)D deficiency is common in pediatric renal transplant recipients and correlates with hyperparathyroidism and short stature. 25(OH)D deficiency may be a modifiable risk factor for hypertension in transplant recipients. Further studies are required to test if routine supplementation with ergo or cholecalciferol is safe and effective in children after renal transplantation. PMID- 21643945 TI - 2D/3D image fusion of X-ray mammograms with breast MRI: visualizing dynamic contrast enhancement in mammograms. AB - PURPOSE: Breast cancer is the most common cancer among women. The established screening method to detect breast cancer is X-ray mammography. Additionally, MRI is used for diagnosis in clinical routine. Due to complementary diagnostic information, both modalities are often read in combination. Yet, the correlation is challenging due to different dimensionality of images and different patient positioning. In this paper, we describe a method to fuse X-ray mammograms with DCE-MRI. The present study was conducted to evaluate the feasibility of the approach. METHODS: For the combination of information from both modalities, the images have to be registered using a compression simulation based on a patient specific biomechanical model. The registered images can be compared directly. The contrast enhancement in the DCE-MRI volume is evaluated using parametric enhancement maps. A projection image of the contrast enhancement is created. The image fusion combines it with X-ray mammograms for intuitive multimodal diagnosis. RESULTS: The image fusion was evaluated using 11 clinical datasets. For 10 of 11 datasets, a good accuracy of the image registration was achieved. The overlap of contrast-enhanced regions with marked lesions in the mammogram is 61%. Lesions are clearly differentiable from surrounding tissue by the DCE-MRI projection in 10 of 11 cases. CONCLUSION: The described preliminary results are promising, thus we expect the visualization of quantitative information from dynamic MRI together with mammograms to be beneficial for multimodal diagnosis. Because of the use of clinical standard modalities, no additional image acquisition is needed. PMID- 21643946 TI - Horizon stabilized--dynamic view expansion for robotic assisted surgery (HS-DVE). AB - PURPOSE: New surgical approaches based on natural orifice transluminal surgery (NOTES) have the potential to further decrease morbidity and hospital stay. However, a number of key challenges have been identified preventing its clinical adoption, including inadequate instrument design and spatial disorientation. Furthermore, retroflexion, missing fixed anatomical references, and limited field of-view are key factors contributing to disorientation in NOTES. METHODS: A hybrid approach of integrated orientation sensing and real-time vision processing is proposed to restore orientation cues for improved surgical navigation. The distal tip of an articulated robotic endoscope is equipped with an inertial measurement unit (IMU) enabling video images to be reoriented and stabilized with respect to the horizon. This is performed by measuring the direction of gravity in relation to the cameras. Dynamic view expansion is used to increase the field of-view of the endoscope. The method registers past video images to the current image and creates an enlarged visualization of the anatomy through simultaneous localization and mapping (SLAM). RESULTS: The clinical potential of the system is demonstrated on a NOTES appendectomy procedure performed on the NOSsE phantom. This involves an articulated robotic endoscope navigating to visualize the appendix while retroflexed. The horizon stabilization is additionally evaluated quantitatively against known ground truth. CONCLUSIONS: The combination of horizon stabilization and dynamic view expansion presents a realistic approach for reintroducing orientation and navigation cues during NOTES. The platform allows real-time implementation, which is an important prerequisite for further clinical evaluation. PMID- 21643947 TI - Subcoracoid impingement syndrome: a painful shoulder condition related to different pathologic factors. AB - Subcoracoid impingement syndrome represents a rare cause of shoulder pain. To date, there are a few papers in literature that have addressed specifically the subcoracoid impingement. We reviewed 13 consecutive patients suffering from this syndrome who underwent an arthroscopic treatment. There were 4 men and 9 women with a mean age of 45 years (range, 23-58 years). The diagnosis of subcoracoid impingement was carried out on the basis of clinical examination and magnetic resonance imaging finding. Arthroscopic surgery consisted of a coracoplasty alone in 2 patients, coracoplasty and acromioplasty in 2 patients, coracoplasty and subscapularis tendon repair in 4 patients, and in the last 5 patients no coracoplasty was done and surgery consisted in treating a minor shoulder instability. Patients were reviewed at a mean follow-up of 2.4 +/- 0.7 years. We evaluated the difference between preoperative and final postoperative range of motion, VAS, UCLA, SST and Constant score using a Student's t test. At follow-up, we observed a significant improvement in range of motion and shoulder scores; moreover, clinical findings of subcoracoid impingement were negative in all patients. Different pathological shoulder conditions can be responsible for a subcoracoid impingement that can be primary or secondary to factors different from mechanic attrition against the coracoid because of its morphology. In case of primary impingement, coracoplasty is a good treatment to relieve clinical symptoms. In patients suffering from an associated minor shoulder instability with MGHL capsulolabral lesion, surgical treatment of this lesion without coracoplasty led to the improvement in symptoms. PMID- 21643948 TI - Replication study of novel risk variants in six genes with type 2 diabetes and related quantitative traits in the Han Chinese lean individuals. AB - To replicating the associations of type 2 diabetes (T2D) and six novel reported variants in Han Chinese lean individuals of first episode T2D, a total of six high risk single nucleotide polymorphisms (SNPs) from the BCL11A, DUSP9, IRS1, CENTD2, ADRA2A, and CDKAL1 genes were examined. Candidate six SNPs were genotyped in 761 T2D patients and 433 control subjects, and associations between the six SNPs and Body Mass Index (BMI), Fasting Plasma Glucose (FPG) and Two Hours Oral Glucose Tolerance Test (2hOGTT) were also investigated. CDKAL1 provided the strongest evidence for replication, where rs7754840 was associated with T2D (odds ratio = 1.54, per copy of the risk C allele, P = 8.10 * 10(-7)). SNP rs5945326 at DUSP9 showed modest significance (odds ratio = 0.81, per copy of the protective G allele, P = 0.02). After adjusting the confounders of age, gender and BMI, the above results remain significant for both rs7754840 (P < 1.0 * 10(-4)) and rs5945326 (P = 0.043) respectively. After correcting for multiple testing, however, only the association between T2D and rs7754840 at CDKAL1 (P < 1*10(-4)) remains significant. In addition, the risk C allele of CDKAL1 rs7754840 was significantly associated with increased FPG levels (P = 3.8 * 10(-4)). The association between genetic variant in CDKAL1 gene was detected in the Han Chinese lean individuals. The correlation between rs7754840-C allele and increased FPG levels is consistent with the potential function of CDKAL1 gene in pancreatic islets. PMID- 21643949 TI - Isolation, characterization, and function analysis of a flavonol synthase gene from Ginkgo biloba. AB - Flavonols are produced by the desaturation of dihydroflavanols, which is catalyzed by flavonol synthase (FLS). FLS belongs to the 2-oxoglutarate iron dependent oxygenase family. The full-length cDNA and genomic DNA sequences of the FLS gene (designated as GbFLS) were isolated from Ginkgo biloba. The full-length cDNA of GbFLS contained a 1023-bp open reading frame encoding a 340-amino-acid protein. The GbFLS genomic DNA had three exons and two introns. The deduced GbFLS protein showed high identities with other plant FLSs. The conserved amino acids (H-X-D) ligating ferrous iron and residues (R-X-S) participating in 2 oxoglutarate binding were found in GbFLS at similar positions like other FLSs. GbFLS was found to be expressed in all tested tissues including roots, stems, leaves, and fruits. Expression profiling analyses revealed that GbFLS expression was induced by all of the six tested abiotic stresses, namely, UV-B, abscisic acid, cold, sucrose, salicylic acid, and ethephon, consistent with the in silico analysis results of the promoter region. The recombinant protein was successfully expressed in the E. coli strain BL21 (DE3) with a pET-28a vector. The in vitro enzyme activity assay by high performance liquid chromatography indicated that recombinant GbFLS protein could catalyze the formation of dihydrokaempferol to kaempferol and the conversion of kaempferol from naringenin, suggesting that GbFLS is a bifunctional enzyme within the flavonol biosynthetic pathway. PMID- 21643950 TI - MicroRNAs in brain development and degeneration. AB - microRNAs are short, non-coding RNAs, that exert a posttranscriptional control on protein synthesis by mRNA interference. They are involved in normal and pathological embryologic development, as well as in adult life pathology, from myocardial infarction to cancer. There are several brain-specific species of microRNA, showing time-dependent pattern of expression, selectivity for neuronal population, significant roles in correct cellular differentiation and system development. The growing interest in microRNAs extended also in the area of neurodegeneration, some of brain-restricted microRNAs being reported to associate with disorders such as Alzheimer's disease, Parkinson's disease or Huntington's disease. The microRNAs research in the last 3 years offered a considerable amount of information that needs to be integrated in the vast machinery of cellular biology. PMID- 21643951 TI - Characterisation and expression analysis of the Atlantic halibut (Hippoglossus hippoglossus L.) cytokines: IL-1beta, IL-6, IL-11, IL-12beta and IFNgamma. AB - Genes encoding the five Atlantic halibut (Hippoglossus hippoglossus L.) cytokines; interleukin (IL)-1beta, IL-6, IL-11b, IL-12betac, and interferon (IFN) gamma, were cloned and characterised at a molecular level. The genomic organisation of the halibut cytokine genes was similar to that seen in mammals and/or other fish species. Several mRNA instability motifs were found within the 3'-untranslated region (UTR) of all cytokine cDNA sequences. The putative cytokine protein sequences showed a low sequence identity with the corresponding homologues in mammals, avian and other fish species. Nevertheless, important structural features were presumably conserved such as the presence, or absence in the case of IL-1beta, of a signal peptide, secondary structure and family signature motifs. The relative expression pattern of the cytokine genes was analyzed in several halibut organs, revealing a constitutive expression in both lymphoid and non-lymphoid organs. Interestingly, the gills showed a relatively high expression of IL-1beta, IL-12betac and IFNgamma. The real time RT-PCR data also showed that the mRNA level of IL-1beta, IL-6, IL-12betac and IFNgamma was high in the thymus, while IL-11b was relatively highly expressed in the posterior kidney and posterior gut. Moreover, the halibut brain showed a relatively high level of IL-6 transcripts. Anterior kidney leucocytes in vitro stimulated with imiquimod showed a significant increase in mRNA level of the five halibut cytokine genes. The sequence and characterisation data presented here will be useful for further investigation of both innate and adaptive immune responses in halibut, and be helpful in the design of vaccines for the control of various infectious diseases. PMID- 21643952 TI - Thymidylate synthase and methionine synthase polymorphisms are not associated with susceptibility to childhood acute lymphoblastic leukemia in Kurdish population from Western Iran. AB - In order to determine the influence of polymorphism in thymidylate synthase (TS 28-bp repeat) and methionine synthase (MS A2756G) genes on the susceptibility to acute lymphoblastic leukemia (ALL), 73 children with ALL and 128 age and sex matched unrelated healthy individuals from the Kermanshah Province of Iran were screened. The genotyping of TS 28-bp repeat and MS A2756G polymorphisms were performed by polymerase chain reaction (PCR) and PCR-RFLP, respectively. The frequency of TS 2R allele in patients and controls were 41.5 and 38%, respectively (Odds ratios (OR) = 1.13, 95%CI 0.73-1.74, P = 0.56). The allelic frequency of G allele of MS was higher (25%) in patients compared with healthy subjects (23%) (OR = 1.09, 95%CI 0.67-1.75, P = 0.71). Considering MS AA and TS 3R3R genotypes as reference indicated that individuals with MS GG + TS 2R2R genotypes have 1.3-fold increase in the risk of ALL (OR = 1.3, 95%CI 0.6-2.7, P = 0.5). Our results showed that neither TS 28-bp repeat nor MS A2756G polymorphisms are risk factors for susceptibility to ALL in Western Iran. PMID- 21643953 TI - Nucleofection induces non-specific changes in the metabolic activity of transfected cells. AB - Transfection has become an everyday technique widely used for functional studies in living cells. The choice of the particular transfection method is usually determined by its efficiency and toxicity, and possible functional consequences specific to the method used are normally overlooked. We describe here that nucleofection, a method increasingly used because of its convenience and high efficiency, increases the metabolic rate of some cancer cells, which can be misleading when used as a measure of proliferation. Moreover, nucleofection can alter the subcellular expression pattern of the transfected protein. These undesired effects are independent of the transfected nucleic acid, but depend on the particular cell line used. Therefore, the interpretation of functional data using this technology requires further controls and caution. PMID- 21643954 TI - Gene expression profiling in zebrafish embryos exposed to diclofenac, an environmental toxicant. AB - Pharmaceuticals are continually released in the environment and therefore pollution from drugs is a pressing problem in the environment. Diclofenac, 2 [(2,6-dichlorophenyl)amino]phenylacetic acid is a FDA approved non-steroidal anti inflammatory drug (NSAID) for the treatment of inflammation. This pharmaceutical has been found as pollutant in superficial waters. Danio rerio (zebrafish) embryo has been used as a model organism for acute pollutant toxicity tests in order to identify morphological alterations in development and death rate. Through the combination of mRNA differential display and quantitative Real Time experiments, we analyzed the alterations of gene expression in zebrafish embryos left to develop in the presence of diclofenac and thereby assess the molecular mechanism involved in ecotoxicity of diclofenac polluted waters. This approach, in embryos exposed to 1.25 mg/l drug for 48 h, allowed identifying 36 different genes, with both known and unknown functions, whose transcription is differentially regulated. The identity and ontological classification of these genes is presented. The wide variety of functional classes of transcripts isolated in this screen reflects the diverse spectrum of influences operating across diclofenac exposure. Of these 36 genes, several have been selected for detailed quantitative Real Time analysis to validate the screen. Our results, for the first time, provide an insight into some of the varied and novel molecular networks following zebrafish exposure to diclofenac polluted waters. PMID- 21643955 TI - Microarray analysis of differentially expressed background genes in rats following hemorrhagic shock. AB - To uncover the contribution of the diversity of the genetic backgrounds to the pathogenesis of hemorrhagic shock, we employed male Sprague-Dawley rats to establish a controlled 2.5 ml/100 g total body weight fixed-volume hemorrhagic shock and left lobular hepatectomy model. RNA was isolated from the liver samples taken from the rats (survival group: rats survived over 24 h after shock; and dead group: rats died within 1 h after shock, n = 3 per group), and subjected to microarray using the illumina(TM) chips for rat cDNA (27,342 genes, >700,000 probes). The results demonstrated that the rats had about 50% survival rate and 100 genes were identified differentially expressed in the two groups. Of these genes, 47 genes were up-regulated and 53 genes down-regulated. Real-time PCR confirmed the differential expression for Aldh1a1, Aldh1a7, Aoc3, Cyp26al, Hdc and Ephx2 genes. Pathway analysis revealed that these genes are involved in circadian rhythm, beta-Alanine metabolism, histidine metabolism, biosynthesis of unsaturated fatty acids, glycine, serine and threonine metabolism, vitamin B6 metabolism, as well as arginine and proline metabolism. Therefore, our study provided a global molecular view on the contribution of genetic backgrounds to the response to hemorrhagic shock. PMID- 21643956 TI - CXCL12 G801A polymorphism and breast cancer risk: a meta-analysis. AB - The G801A polymorphism in the CXCL12 gene has been implicated in breast cancer risk. However, the published findings are inconsistent. We therefore performed a meta-analysis to investigate this relationship. Odds ratios (ORs) and 95% confidence intervals (CIs) were used to assess the strength of the association. The pooled ORs were performed for codominant model, dominant model, and recessive model, respectively. Five published case-control studies, including 1,058 breast cancer cases and 1,023 controls were identified. No study had a deviation from the Hardy-Weinberg equilibrium (HWE) in controls. We found that the CXCL12 G801A (rs1801157) polymorphism was associated with a significantly increased risk of breast cancer risk when all studies were pooled into the meta-analysis (codomiant model: AA versus GG, OR = 1.64, 95% CI = 1.16-2.33; GA versus GG, OR = 1.42, 95% CI = 1.18-1.71; dominant model: AA/GA versus GG, OR = 1.44, 95% CI = 1.21-1.72). Furthermore, Egger's test did not show any evidence of publication bias (P > 0.05 for the dominant model). In conclusion, the results suggest that the CXCL12 G801A polymorphism may be a low-penetrant risk factor for developing breast cancer. PMID- 21643957 TI - Molecular cloning and expression analysis of a RGA-like gene responsive to plant hormones in Brassica napus. AB - DELLA proteins are negative regulators of GA-induced growth. DELLA protein family is characterized by a DELLA domain essential for GA-dependent proteasomal degradation of DELLA repressors. A full-length cDNA encoding a putative DELLA protein with high sequence homology to Arabidopsis thaliana RGA (AtRGA), designated as BnRGA, was isolated from Brassica napus. The full-length cDNA of BnRGA contained a 1,740 bp open reading frame (ORF) encoding a precursor protein of 579 amino acid residues. Comparative and bioinformatics analyses revealed that BnRGA showed a high degree of homology with DELLA proteins and contained the DELLA domain, TVHYNP domain, VHIID domain and RVER domain. Using real-time PCR, the expression patterns of BnRGA and two our previously isolated genes, BnGID1a and BnSLY1 in B. napus, were analyzed by adding exogenous gibberellins acid-3 (GA(3)), GA biosynthetic inhibitor paclobutrazol (PAC) and abscisic acid (ABA). The results showed that the expression of BnGID1a and BnSLY1 was down-regulated after treated by GA(3) and induced by PAC and ABA. These results suggest that the expression of BnGID1a and BnSLY1 may be negatively regulated by the level of endogenous GA in B. napus. Moreover, BnRGA was not significantly regulated by GA(3), PAC and ABA in the low concentrations. These suggest that GA-GID1-SCF DELLA complex may have a mechanism of self-regulation, thereby preserving the stability of the expression level of BnRGA in B. napus. PMID- 21643958 TI - Transcriptomic analysis of the housefly (Musca domestica) larva using massively parallel pyrosequencing. AB - To explore the transcriptome of Musca domestica larvae and to identify unique sequences, we used massively parallel pyrosequencing on the Roche 454-FLX platform to generate a substantial EST dataset of this fly. As a result, we obtained a total of 249,555 ESTs with an average read length of 373 bp. These reads were assembled into 13,206 contigs and 20,556 singletons. Using BlastX searches of the Swissprot and Nr databases, we were able to identify 4,814 contigs and 8,166 singletons as unique sequences. Subsequently, the annotated sequences were subjected to GO analysis and the search results showed a majority of the query sequences were assignable to certain gene ontology terms. In addition, functional classification and pathway assignment were performed by KEGG and 2,164 unique sequences were mapped into 184 KEGG pathways in total. As the first attempt on large-scale RNA sequencing of M. domestica, this general picture of the transcriptome can establish a fundamental resource for further research on functional genomics. PMID- 21643959 TI - Impact of DNA repair genes polymorphism (XPD and XRCC1) on the risk of breast cancer in Egyptian female patients. AB - The genes involved in DNA repair system play a crucial role in the protection against mutations. It has been hypothesized that functional deficiencies in highly conserved DNA repair processes resulting from polymorphic variation may increase genetic susceptibility to breast cancer (BC). The aim of the present study was to evaluate the association of genetic polymorphisms in 2 DNA repair genes, XPD (Asp312Asn) and XRCC1 (A399G), with BC susceptibility. We further investigated the potential combined effect of these DNA repair variants on BC risk. Both XPD (xeroderma pigmentosum group D) and XRCC1 (X-ray repair cross complementing group 1) polymorphisms were characterized in 100 BC Egyptian females and 100 healthy women who had no history of any malignancy by amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) method and PCR with confronting two-pair primers (PCR-CTPP), using DNA from peripheral blood in a case control study. Our results revealed that the frequencies of AA genotype of XPD codon 312 polymorphism were significantly higher in the BC patients than in the normal individuals (P <= 0.003), and did not observe any association between the XRCC1 Arg399Gln polymorphism and risk of developing BC. Also, no association between both XPD Asp312Asn and XRCC1 A399G polymorphisms and the clinical characteristics of disease. Finally, the combination of AA(XPD) + AG(XRCC1) were significantly associated with BC risk. Our results suggested that, XPD gene is an important candidate gene for susceptibility to BC. Also, gene-gene interaction between XPD(AA) + XRCC1(AG) polymorphism may be associated with increased risk of BC in Egyptian women. PMID- 21643960 TI - Molecular cloning, tissue distribution and ontogenetic expression of Xiang pig chemerin and its involvement in regulating energy metabolism through Akt and ERK1/2 signaling pathways. AB - Chemerin, as a new member of adipokines family, is highly expressed in adipose tissue in rodent and its expression increases with obesity. Moreover, chemerin has been reported to have significant relationship with metabolic syndrome and insulin sensitivity. Here, the gene encoding chemerin from Xiang pig was cloned. The open reading frame of this cDNA encodes 163 deduced amino acid residues. The putative protein has a N-terminal signaling peptide and a nuclear localization signal profile which are highly conserved among the vertebrate orthologs. Both chemerin and chemerinR are highly expressed in lung, kidney and small intestine in adult Xiang pig. Besides these tissues, chemerin is abundant in liver and backfat, and chemerinR is abundant in spleen and skeletal muscle. We also investigated the age-dependent expression of chemerin in suckling Xiang piglets in various tissues, which showed an interaction between age and segments in abundance of chemerin and chemerinR from day 1 to day 21. For chemerinR, it was abundant in skeletal muscle of both adult and fetal Xiang pig. Further, we treated differentiated C2C12 cells with chemerin. The result showed that chemerin regulated energy metabolism partly through Akt and ERK1/2 signaling pathway. Taken together, our findings provide basic molecular information for the deeper investigation on the function of chemerin. PMID- 21643961 TI - The effect of selegiline on total scavenger capacity and liver fat content: a preliminary study in an animal model. AB - Selegiline is a selective irreversible inhibitor of the B-type of monoamine oxidase (MAO-B). The spectrum of its pharmacological activity is wide, possesses antioxidant, antiapoptotic and neuroprotective properties and, additionally, we found it is effective on the total scavenger capacity (TSC), and the regulation of fat content in rat liver kept on lipid-rich diet. Our aim was to clarify whether the oral treatment with selegiline is protective on oxidative damage of Sprague-Dawley adult rats in vivo. Four groups of rats (five animals in a group) were examined: (1) lipid-rich diet, (2) normal rat food, (3) lipid-rich diet + selegiline and (4) normal rat food + selegiline. Selegiline solution (2.5 ug/ml) was supplied with the drinking water, which was freely available for the animals. Regarding the drinking habit of the rats (20-30 ml/day), the daily dose was roughly equal with that used in the human therapy (5-10 mg/day). TSC was determined both at the beginning (0 day) and at the end of the study (28 days), when the blood samples were taken for chemiluminometric assay. Fat content of the liver was determined in the freshly frozen tissue by Sudan staining. TSC was increased in both the selegiline-treated groups. Selegiline treatment prevented the increase of liver fat in the group fed with lipid-rich diet. Our results led us to the conclusion that prolonged selegiline administration can raise the antioxidant capacity of the animals and prevents the accumulation of fat in their livers. PMID- 21643962 TI - Pediatric parenting stress and its relation to depressive symptoms and fear of hypoglycemia in parents of young children with type 1 diabetes mellitus. AB - Parents of young children with type 1 diabetes (T1DM) maintain full responsibility for their child's daily diabetes self-care and thus may be vulnerable to experiencing parenting stress. This study examined several psychological correlates of pediatric parenting stress in parents of young children with T1DM. Parents of 39 young children with T1DM (ages 2-7 years) completed measures of pediatric parenting stress, mealtime behavior problems, depressive symptoms, and fear of hypoglycemia. For parents of young children, higher stress frequency and difficulty were associated with higher parental depressive symptoms and fear. Regression analyses identified that 58% of the variance in stress frequency was associated with parental depressive symptoms. For stress difficulty, 68% of the variance was associated with parental depressive symptoms and fear. Pediatric parenting stress is common in parents of young children with T1DM. Stress and the psychological correlates measured in this study are amenable to intervention and should be regularly assessed in parents of young children with T1DM. PMID- 21643963 TI - Technology complementing military behavioral health efforts at tripler army medical center. AB - The purpose of this article is to provide a short narrative on the ways that behavioral health professionals and their patients are currently benefitting from the use of technology. Examples stem from applications of technology to patients/research participants at the Tripler Army Medical Center. The paper also discusses how current use of this technology has made it possible to serve individuals in their own cultural environment, providing a cost-effective means of providing mental health services. PMID- 21643964 TI - The dilemma, causes and approaches to avoid recurrent hospital readmissions for patients with chronic heart failure. AB - Heart failure is a progressive illness that carries significant morbidity and mortality. This highly prevalent illness leads to frequent, costly hospitalizations with approximately 50% of patients being readmitted within 6 months of initial hospitalization. While rehospitalization has been extensively studied in the past, little progress has been made in terms of reducing readmission rates of heart failure patients in the last decade despite increasing costs with impending resource limitations. We discuss disease-centered, physician centered, and patient-centered factors that lead to rehospitalization as well as community/resource availability factors that contribute to rehospitalization of patients suffering from chronic heart failure. In addition, predictors of hospitalization and interventions that reduce hospitalization will be critically evaluated. With a complete understanding of heart failure rehospitalization, we hope the future holds more effective ways to prevent heart failure progression and thus rehospitalization, improved risk-stratification models to identify patients high-risk for rehospitalization, and sustained interventions that are customized according to the etiology of the clinical decline of heart failure patients that ultimately results in frequent rehospitalizations. PMID- 21643965 TI - Occurrence of halogenated contaminants in fish from selected river localities and ponds in the Czech Republic. AB - The occurrence of organohalogenated compounds including major persistent chlorinated pollutants, such as polychlorinated biphenyls (PCBs) and DDT and its metabolites, brominated flame retardants (BFRs), represented by polybrominated diphenylethers (PBDEs) and hexabromocyclododecane (HBCD), together with currently widely discussed perfluorinated compounds (PFCs), mainly perfluorooctane sulfonic acid was monitored in several fish species collected from Czech rivers. Eleven sampling locations in highly industrialized areas were chosen. In addition, wild species of 14 farmed fish (grown in dedicated ponds) were also analysed. With respect to the contamination in different areas, chlorinated chemicals were dominant. PCBs and DDTs ranged from 4.8 to 211 and 2 to 791 MUg/kg wet weight, respectively. Concentrations of BFRs and PFCs were significantly lower and ranged from 0.6 to 10.2 and 0.9 to 62 MUg/kg wet weight, respectively. The highest levels of target analyte groups were found in fish muscle tissue in localities situated on the lower part of the Elbe River: Levels of DDT, HCB, PBDEs, and perfluorooctane sulfonic acid (PFOS) were as high as 791, 77.6, 14.4 and 193 MUg/kg wet weight, respectively, in Usti nad Labem and in the sample originated from the confluence of Elbe with Bilina River; a sum of PCBs at a level of 211 MUg/kg was detected. PMID- 21643966 TI - 1,4-Dioxane enhances properties and biocompatibility of polyanionic collagen for tissue engineering applications. AB - Polyanionic collagen obtained from bovine pericardial tissue submitted to alkaline hydrolysis is an acellular matrix with strong potential in tissue engineering. However, increasing the carboxyl content reduces fibril formation and thermal stability compared to the native tissues. In the present work, we propose a chemical protocol based on the association of alkaline hydrolysis with 1,4-dioxane treatment to either attenuate or revert the drastic structural modifications promoted by alkaline treatments. For the characterization of the polyanionic membranes treated with 1,4-dioxane, we found that (1) scanning electron microscopy (SEM) shows a stronger reorientation and aggregation of collagen microfibrils; (2) histological evaluation reveals recovering of the alignment of collagen fibers and reassociation with elastic fibers; (3) differential scanning calorimetry (DSC) shows an increase in thermal stability; and (4) in biocompatibility assays there is a normal attachment, morphology and proliferation associated with high survival of the mouse fibroblast cell line NIH3T3 in reconstituted membranes, which behave as native membranes. Our conclusions reinforce the ability of 1,4-dioxane to enhance the properties of negatively charged polyanionic collagen associated with its potential use as biomaterials for grafting, cationic drug- or cell-delivery systems and for the coating of cardiovascular devices. PMID- 21643967 TI - Synthesis of novel 6-triazologlycolipids via click chemistry and their preliminary cytotoxicity assessments. AB - Series of novel 6-triazole-linked galacto- or glucolipids were efficiently synthesized from O-benzylated sugar azides and various lipid alkynes via Cu(I) catalyzed azide-alkyne 1,3-dipolar cycloaddition (click chemistry) followed by hydrogenolysis with PdCl(2)/H(2). Subsequent MTT (3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide) assay toward a panel of human cancer cell lines revealed that these triazologlycolipids possess low-to-modest toxicity on A549 (lung), MCF-7 (breast), HeLa (cervix), and HepG2 (liver). Furthermore, both the carbon chain length at the lipid end and the epimeric identity of the glycosyl moiety were determined to impact their corresponding bioactivity. PMID- 21643968 TI - 1H, 13C and 15N resonance assignments of human muscle acylphosphatase. AB - Human muscle acylphosphatase (mAcP) is an enzyme with a ferrodoxin-like topology whose primary role is to hydrolyze the carboxyl-phosphate bonds of acylphosphates. The protein has been widely used as a model system for elucidating the molecular determinants of protein folding and misfolding. We present here the full NMR assignments of the backbone and side chains resonances of mAcP complexed with phosphate, thus providing an important resource for future solution-state NMR spectroscopic studies of the structure and dynamics of this protein in the contexts of protein folding and misfolding. PMID- 21643969 TI - 1H, 13C, and 15N resonance assignment of the SPFH domain of human stomatin. AB - Stomatin, a 288-residue protein, is a component of the membrane skeleton of red blood cells (RBCs), which helps to physically support the membrane and maintains its function. In RBCs, stomatin binds to the glucose transporter GLUT-1 and may regulate its function. Stomatin has a stomatin/prohibitin/flotillin/HflK (SPFH) domain at the center of its polypeptide chain. There are 12 SPFH domain containing proteins, most of which are localized at the cellular or subcellular membranes. Although the molecular function of the SPFH domain has not yet been established, the domain may be involved in protein oligomerization. The SPFH domain of the archaeal stomatin homolog has been shown to form unique oligomers. Here we report the (15)N, (13)C, and (1)H chemical shift assignments of the SPFH domain of human stomatin [hSTOM(SPFH)]. These may help in determining the structure of hSTOM(SPFH) in solution as well as in clarifying its involvement in protein oligomerization. PMID- 21643970 TI - NMR resonance assignment of the autoimmunity protein SpaI from Bacillus subtilis ATCC 6633. AB - Bacillus subtilis ATCC 6633 produces the lipid II targeting lantibiotic subtilin. For self-protection these gram-positive bacteria express a cluster of four self immunity proteins named SpaIFEG. SpaI is a 16.8 kDa lipoprotein which is attached to the outside of the cytoplasmic membrane via a covalently linked diacylglycerol anchor. Together with the ABC-transporter SpaFEG, SpaI protects the membrane from subtilin insertion and there is evidence for a direct interaction of SpaI with subtilin. As a prerequisite for further structural studies of SpaI and the SpaI/subtilin complex we report here the full (1)H, (15)N, (13)C chemical shift assignment for a stable 14.9 kDa C-terminal fragment of SpaI. PMID- 21643971 TI - The role of RANK/RANKL/OPG signalling pathways in osteoclastogenesis in odontogenic keratocysts, radicular cysts, and ameloblastomas. AB - The aim of this study was to evaluate the immunohistochemical expression of molecules involved in osteoclastogenesis, including the receptor activator of nuclear factor kappa B (RANK), RANK ligand (RANKL) and osteoprotegerin (OPG) in odontogenic keratocysts (OKCs), which has been named as a keratocystic odontogenic tumour by the WHO, and compare their expression with radicular cysts and ameloblastomas. RANK is a member of tumour necrosis factor receptor family and it is activated by RANK ligand. OPG binds to RANKL and inactivates it. The imbalance of these factors could cause the differential bone resorption activity in some diseases and tumours. The expression of these molecules was evaluated in ameloblastomas (n = 20), OKCs (n = 20), and radicular cysts (n = 20) by immunohistochemistry. Immunohistochemical reactivity for RANK, RANKL, and OPG was detected in neoplastic and nonneoplastic epithelium and connective tissue cells. RANK showed the greatest expression in OKCs followed by ameloblastomas, with the lowest expression seen in radicular cysts. Expression of RANKL was detected in all lesions and no significant differences were observed between groups. OPG was expressed very low in all groups. In the stroma, the number of RANK positive cells was higher in OKCs when compared with ameloblastomas and radicular cysts but radicular cyst had higher numbers of RANKL positive cells in the stroma than ameloblastomas. The molecular system of RANK/RANKL/OPG is variably expressed in OKCs, radicular cysts, and ameloblastomas and this system may be involved in the osteoclastogenic mechanisms in OKCs and ameloblastomas. Advanced studies could further clarify the role of RANK, RANKL, and OPG in mediating tumour associated bone osteolysis. PMID- 21643972 TI - ROCK inhibition prevents fetal serum-induced alteration in structure and function of organ-cultured mesenteric artery. AB - Chronic treatment with fetal bovine serum (FBS) causes contractility reduction, morphological alteration and DNA synthesis in organ-cultured vascular tissues. Here, we tested the hypothesis that chronic inhibition of ROCK has a protective effect on FBS-induced alterations in small arteries. Rabbit mesenteric arterial rings were cultured in FBS-supplemented culture medium with or without Y-27632, a reversible ROCK inhibitor. Chronic Y-27632 treatment prevented FBS-induced gradual arterial constriction, wall thickening, reduced contractility, and increased ROCK-specific MYPT1 Thr853 phosphorylation. Treatment with Y-27632 also prevented decreased eNOS mRNA expression, and reduced acetylcholine-induced relaxation. Sudden application of Y-27632 to pre-cultured rings reduced MYPT1 phosphorylation and re-widened the constricted rings. Chronic treatment with Y 27632, however, rather augmented than reduced the FBS-induced RhoA over expression, also increased ROCK1 and MYPT1 expression and averted the FBS-induced reduction of MLC expression, suggesting a compensation of inhibited RhoA/ROCK activity. Sudden removal of Y-27632 caused a rebound in MYPT1 phosphorylation and vasoconstriction in rabbit mesenteric artery. To test which ROCK isoform has greater involvement in FBS-induced contraction, haploinsufficient Rock1+/- and Rock2+/- mouse mesenteric arterial rings were subjected to organ-culture. FBS induced contraction and RhoA over-expression in either heterozygous animal was not different from wild-type animals. These results suggest that FBS-induced contraction is mediated by up-regulation of RhoA and subsequent activation of ROCK. In conclusion, chronic ROCK inhibition produces some effects that protect against FBS-stimulated vasoconstriction and remodeling. There are also negative effects that a sudden withdrawal of ROCK inhibitor might cause a stronger vasoconstriction than before it was used. PMID- 21643973 TI - Loop 1 dynamics in smooth muscle myosin: isoform specific differences modulate ADP release. AB - Isoforms of the smooth muscle (SM) myosin motor domain differ in the presence or absence of a seven amino acid insert in a flexible surface loop spanning the nucleotide-binding pocket known as Loop 1. The presence of this insert leads to a two-fold increase in actin sliding velocity and ADP release rate between these isoforms, although the effect of Loop 1 on the kinetics of ADP release remains unclear. To further investigate the role of the Loop 1 insert in modulating ADP release in SM myosin we have inserted a single tryptophan residue into Loop 1 of both isoforms as a probe of local structural dynamics. By monitoring the dynamics of Loop 1 in relation to the release of ADP we have observed a unique movement of Loop 1 in the inserted isoform, preceding nucleotide release, which is absent in the non-inserted isoform. This movement is sequence dependent as alanine replacement of the insert residues abolishes the transition and slows ADP release. Thus movement of Loop 1 is a critical factor in increasing the ADP release rate in the inserted faster isoform of SM myosin. PMID- 21643974 TI - Prediction of problematic wine fermentations using artificial neural networks. AB - Artificial neural networks (ANNs) have been used for the recognition of non linear patterns, a characteristic of bioprocesses like wine production. In this work, ANNs were tested to predict problems of wine fermentation. A database of about 20,000 data from industrial fermentations of Cabernet Sauvignon and 33 variables was used. Two different ways of inputting data into the model were studied, by points and by fermentation. Additionally, different sub-cases were studied by varying the predictor variables (total sugar, alcohol, glycerol, density, organic acids and nitrogen compounds) and the time of fermentation (72, 96 and 256 h). The input of data by fermentations gave better results than the input of data by points. In fact, it was possible to predict 100% of normal and problematic fermentations using three predictor variables: sugars, density and alcohol at 72 h (3 days). Overall, ANNs were capable of obtaining 80% of prediction using only one predictor variable at 72 h; however, it is recommended to add more fermentations to confirm this promising result. PMID- 21643975 TI - Partial nitrification and nutrient removal in intermittently aerated sequencing batch reactors treating separated digestate liquid after anaerobic digestion of pig manure. AB - The performance of an intermittently aerated sequencing batch reactor (IASBR) technology was investigated in achieving partial nitrification, organic matter removal and nitrogen removal from separated digestate liquid after anaerobic digestion of pig manure. The wastewater had chemical oxygen demand (COD) concentrations of 11,540 +/- 860 mg/L, 5-day biochemical oxygen demand (BOD(5)) concentrations of 2,900 +/- 200 mg/L and total nitrogen (TN) concentrations of 4,041 +/- 59 mg/L, with low COD:N ratios (2.9) and BOD(5):COD ratios (0.25). Synthetic wastewater, simulating the separated digestate liquid with similar COD and nitrogen concentrations but BOD(5) of 11,500 +/- 100 mg/L, was also treated using the IASBR technology. At a mean organic loading rate of 1.15 kg COD/(m(3) d) and a nitrogen loading rate of 0.38 kg N/(m(3) d), the COD removal efficiency was 89.8% in the IASBR (IASBR-1) treating digestate liquid and 99% in the IASBR (IASBR-2) treating synthetic wastewater. The IASBR-1 effluent COD was mainly due to inert organic matter and can be further reduced to less than 40 mg/L through coagulation. The partial nitrification efficiency of 71-79% was achieved in the two IASBRs and one cause for the stable long-term partial nitrification was the intermittent aeration strategy. Nitrogen removal efficiencies were 76.5 and 97% in IASBR-1 and IASBR-2, respectively. The high nitrogen removal efficiencies show that the IASBR technology is a promising technology for nitrogen removal from low COD:N ratio wastewaters. The nitrogen balance analysis shows that 59.4 and 74.3% of nitrogen removed was via heterotrophic denitrification in the non-aeration periods in IASBR-1 and IASBR-2, respectively. PMID- 21643976 TI - Effect of nitrogen limitation on enrichment of activated sludge for PHA production. AB - Polyhydroxyalkanoates (PHA) are good candidates to plastics because of their material properties similar to conventional plastics and complete biodegradability. The use of activated sludge can be a cheaper alternative to pure cultures for PHA production. In this study, effect of nitrogen limitation during acclimatization period of biomass on production of polyhydroxyalkanoate was investigated. Activated sludge was selected in two sequencing batch reactors operated with and without nitrogen limitation. Batch tests were performed to examine polymer productions of activated sludges acclimatized to different nitrogen regimes. Responses of biomass to different organic loading rates, organic acids, and carbon to nitrogen (C/N) ratios were studied by determining specific polymer storage rate, polymer storage yield, and sludge polymer content of biomasses. Results obtained from batch experiments showed that concentrations of polymer accumulated by two different sludges increased directly with initial substrate concentration. Observed highest polymer yields for the biomasses enriched with and without nitrogen deficiency were 0.69 g COD PHA g(-1) COD S and 0.51 g COD PHA g(-1) COD S, and corresponding polymer contents of biomasses were 43.3% (g COD PHA g(-1) COD X) and 38.3% (g COD PHA g(-1) COD X), respectively. Polymer yields for both biomasses decreased with substrate shift however, biomass enriched with nitrogen deficiency adapted well to acetate-propionate mixture. The results presented in this study showed that polymer storage ability of biomass was improved more under dynamic conditions with nitrogen deficiency when compared to that without nitrogen deficiency. Limiting ammonia availability during batch experiments also caused higher polymer production by suppressing growth, as well as during enrichment of biomass. PMID- 21643977 TI - Decreased expression of lysosomal alpha-galactosiase A gene in sporadic Parkinson's disease. AB - Parkinson's disease (PD) is a progressive neurodegenerative disease. To date, the causal genes and variants associated with sporadic PD are largely unknown. Accumulating evidence demonstrates that autophagy delivers alpha-syncuclein proteins to lysosome for degradation and dysfunctional autophagy is involved in the PD pathogenesis. We have previously screened a group of lysosomal hydrolases and found that alpha-galactosidase A (GLA) activity is significantly decreased in the peripheral leukocytes of sporadic PD patients. In this study, GLA transcript and protein levels were semi-quantitatively examined. The GLA transcript (P = 0.020) and protein (P = 0.027) levels in the peripheral leukocytes of sporadic PD patients were significantly decreased, compared to age- and sex-matched healthy controls. Furthermore, decreased GLA gene expression levels were strongly associated with sporadic PD (OR 3.33, 95%CI 1.17-9.52, P = 0.024). Therefore, our data suggest that insufficient GLA activity may contribute to the pathogenesis of sporadic PD. The underlying molecular mechanisms remain to be determined. PMID- 21643978 TI - Age-related binding of proximal region of ApoE promoter to nuclear proteins of mouse cerebral cortex. AB - Mouse ApolipoproteinE (ApoE) gene has maximum promoter activity in the proximal region (-212 to +54) which includes different regulatory elements. These elements bind to specific protein factors and influence the expression of genes which are involved in key brain functions that decline with age. As there is no information on the binding of apoE promoter to nuclear proteins as a function of age, we have analyzed the binding of USF, AP1 and one negative element sequence present in ApoE proximal promoter to nuclear proteins of the cerebral cortex of mice of different ages. The findings show the formation of one complex with USF and two complexes with AP1 and negative element. The intensity of these complexes varies with age, indicating differential binding of protein factors to specific elements of apoE promoter, which reflect age-related regulation of apoE -mediated brain functions. PMID- 21643979 TI - A crucial role for IL-6 in the CNS of rats during fever induced by the injection of live E. coli. AB - Interleukin (IL)-1beta, tumor necrosis factor (TNF)-alpha, and IL-6 have been established as important mediators of fever induced by lipopolysaccharide (LPS) from Gram-negative bacteria. Whether these pro-inflammatory cytokines are also important in mediating fever induced by live bacteria remains less certain. We therefore investigated the following: (1) the synthesis of TNF-alpha, IL-1beta, and IL-6 during E. coli-induced fever and (2) the effect of blocking the action of cytokines within the brain on E. coli-induced fever. Body or tail skin temperature (bT or Tsk, respectively) was measured by biotelemetry or telethermometry, every 30 min, during 6 or 24 h. Depending on the number of colony-forming units (CFU) injected i.p., administration of E. coli induced a long-lasting increase in bT of male Wistar rats. The duration of fever did not correlate with the number of CFU found in peritoneal cavity or blood. Because 2.5 * 10(8) CFU induced a sustained fever without inducing a state of sepsis/severe infection, this dose was used in subsequent experiments. The E. coli-induced increase in bT was preceded by a decrease in Tsk, reflecting a thermoregulatory response. TNF-alpha, IL-1beta, and IL-6 were detected at 3 h in serum of animals injected i.p. with E. coli. In the peritoneal exudates, TNF-alpha, IL-1beta, and IL-6 were detected at 0.5 and 3 h after E. coli administration. Moreover, both IL 1beta and IL-6, but not TNF-alpha, were found in the cerebrospinal fluid (CSF) and hypothalamus of animals injected with E. coli. Although pre-treatment (i.c.v., 2 MUl, 15 min before) with anti-IL-6 antibody (anti-IL-6, 5 MUg) reduced E. coli-induced fever, pre-treatment with either IL-1 receptor antagonist (IL 1ra, 200 MUg) or soluble TNF receptor I (sTNFRI, 500 ng) had no effect on the fever response. In conclusion, replicating E. coli promotes an integrated thermoregulatory response in which the central action of IL-6, but not IL-1 and TNF, appears to be important. PMID- 21643980 TI - Incidence of testicular malignancies and correlation to risk factors in a TESE population of subfertile men. AB - INTRODUCTION: In recent years, researchers have postulated a decreasing fertility potential of males and a rising incidence of testicular malignancies. MATERIALS AND METHODS: In this retrospective observational diagnostic multicenter study, 302 patient files of subfertile men whose testes were biopsied for TESE procedure were analysed. All patients referred to reproductive medicine centres in Northern Germany and they were identified by the cycle data collected by the German IVF register. A total of 280 patients (436 cycles) treated for intracytoplasmatic sperm injection after TESE procedures were eligible to be analysed. RESULTS: Our findings: 13.0% overall paternity rate before TESE procedure, 45.9% smokers, maldescensus testis was found on occasion (12.9%), and mumps orchitis previously occurred to 10 patients (3.6%). The tumour incidence rate at the time of testicle biopsy was 1.81% (= 5 pts.). Two of these patients had an anamnesis for maldescensus testis and one patient acquired mumps orchitis in childhood. CONCLUSION: Our data even reflects that tumour patients express an interest in having children after completion of cancer treatment, presenting four patients who had testicular biopsies after a previous malignancy. Moreover, there is evidence suggesting that environmental factors are causative for the trends in occurrence of male reproductive health problems. Within our highly selected population, the testicular tumour incidence rate is 100-fold higher than in a standard population. Supposing that the incidence rate of testicular malignancies among infertile men continues to increase in comparison to the incidence rate of the general male population, one has to count on an incremental number of males suffering from subfertility and testicular tumours. PMID- 21643981 TI - Population biobanks and returning individual research results: mission impossible or new directions? AB - Historically, large-scale longitudinal genomic research studies have not returned individual research results to their participants, as these studies are not intended to find clinically significant information for individuals, but to produce 'generalisable' knowledge for future research. However, this stance is now changing. Commentators now argue that there is an ethical imperative to return clinically significant results and individuals are now expressing a desire to have them. This shift reflects societal changes, such as the rise of social networking and an increased desire to participate in medical decision-making, as well as a greater awareness of genetic information and the increasing ability of clinicians to use this information in health care treatment. This paper will discuss the changes that have prompted genomic research studies to reconsider their position and presents examples of projects that are actively engaged in returning individual research results. PMID- 21643982 TI - Alterations of ATM and CADM1 in chromosomal 11q22.3-23.2 region are associated with the development of invasive cervical carcinoma. AB - To understand the importance of chr11q22.3-23.2 region in the development of cervical cancer, we have studied the genetic and epigenetic alterations of the candidate genes ATM, PPP2R1B, SDHD and CADM1 in cervical intraepithelial neoplasia (CIN) and cervical carcinoma (CACX) samples. Our study revealed low expression and high alterations (methylation/deletion) (55-59%) of ATM and CADM1 genes along with poor patient outcome. The alterations of ATM and CADM1 are associated with the progression of tumor from CIN to Stage I/II, thus implying their role in early invasiveness. The two genes, PPP2R1B and SDHD, lying in between ATM and CADM1, have low frequency of alterations, and majority of the alterations are in CACX samples, indicating that their alterations might be associated with disease progression. Expressions (mRNA/protein) of the genes showed concordance with their molecular alterations. Significant co-alteration of ATM and CADM1 points to their synergic action for the development of CACX. Mutation is, however, a rare phenomenon for inactivation of ATM. Association between the alteration of ATM and CHEK1 and poor survival of the patients having co-alterations of ATM and CHEK1 points to the DNA damage response pathway disruption in development of CACX. Thus, our data suggest that inactivation of ATM-CHEK1-associated DNA damage response pathway and CADM1-associated signaling network might have an important role in the development of CACX. PMID- 21643983 TI - Mary Lyon and the hypothesis of random X chromosome inactivation. AB - The 50th anniversary of Mary Lyon's 1961 Nature paper, proposing random inactivation in early embryonic life of one of the two X chromosomes in the cells of mammalian females, provides an opportunity to remember and celebrate the work of those involved. While the hypothesis was initially put forward by Lyon based on findings in the mouse, it was founded on earlier studies, notably the work of Susumu Ohno; it was also suggested independently by Beutler and colleagues using experimental evidence from a human X-linked disorder, glucose-6-phosphate dehydrogenase deficiency, and has proved to be of as great importance for human and medical genetics as it has for general mammalian genetics. Alongside the hypothesis itself, previous cytological studies of mouse and human chromosomes, and the observations on X-linked mutants in both species deserve recognition for their essential role in underpinning the hypothesis of random X-inactivation, while subsequent research on the X-inactivation centre and the molecular mechanisms underlying the inactivation process represent some of the most outstanding contributions to human and wider mammalian genetics over the past 50 years. PMID- 21643984 TI - Somatic variation and cancer: therapies lost in the mix. AB - Cancer arises as a consequence of mutations in genomes of cancer cells, which over time allow them to proliferate and spread to distant sites. Large-scale sequencing of cancer genomes is revealing an increasing number of potential driver mutations that may allow specific targeting of cancer genes, proteins, and pathways. Comprehensive views of cancer genomes are also revealing enormous heterogeneity of mutation profiles, even among tumours derived from the same organs and having similar pathological characteristics. There are now many examples where mutation profiles observed in tumours have been shown to correlate with clinical features of disease, drug response, and patient outcomes. When ignored, molecular heterogeneity can lead to failures in drug development, as drugs that may have efficacy in subgroups of patients with specific molecular phenotypes may show marginal response when tested in large groups of unselected patients. This article explores issues relevant to the clinical translation of sequence-based mutation profiles in the clinical development of targeted therapies and in the future management of cancer patients. PMID- 21643985 TI - IDH mutation status impact on in vivo hypoxia biomarkers expression: new insights from a clinical, nuclear imaging and immunohistochemical study in 33 glioma patients. AB - Mutations in the gene encoding isocitrate dehydrogenase enzyme isoforms 1 (IDH1) and 2 (IDH2) have recently been identified in a large proportion of glial tumors of the CNS, but their mechanistic role in tumor development remains unclear. Here, we assessed the actual impact of IDH1 and IDH2 mutations in patients harboring WHO grade II and III gliomas. We sequenced IDH1 at codon 132 and IDH2 at codon 172 in 33 patients with WHO grade II and III gliomas who benefited from a preoperative (18)F-FDG positron emission tomography (PET). Immunohistochemical expression of Hypoxia Inducible Factor-1alpha (HIF-1alpha), Carbonic Anhydrase IX (CAIX), Glucose Transporter 1 (GLUT1) and Caspase 3 active form (CASP3) along with the R132HIDH1 mutation was assessed in all cases as well as 1p/19q deletion status and p53 expression. HIF-1alpha expression was found in 15% of IDH-mutated compared to 7.7% of IDH-nonmutated tumors (P = 0.954). Also, GLUT-1 positive staining was found in 5% of IDH-mutated and in 7.1% of IDH-nonmutated tumors (P = 0.794). Finally, CA-IX expression was found in 15% of IDH-mutated and in 7.7% of IDH-nonmutated tumors (P = 0.484). The combined expression of these three hypoxic markers was found in two WHO grade III tumors, one of which was IDH-mutated whereas the other was IDH-nonmutated (P = 0.794). In IDH-mutated tumors, the median SUVmax ratio was 2.24 versus 2.15 in IDH-nonmutated tumors (P = 0.775). Together, these data question the actual relationship between IDH mutation status and in vivo hypoxic biomarkers expression in WHO grade II and III gliomas. PMID- 21643986 TI - Chordoid meningioma in a patient with lymphangioleiomyomatosis. PMID- 21643987 TI - Analysis of DNA repair gene polymorphisms and survival in low-grade and anaplastic gliomas. AB - The purpose of this study was to explore the variation in DNA repair genes in adults with WHO grade II and III gliomas and their relationship to patient survival. We analysed a total of 1,458 tagging single-nucleotide polymorphisms (SNPs) that were selected to cover DNA repair genes, in 81 grade II and grade III gliomas samples, collected in Sweden and Denmark. The statistically significant genetic variants from the first dataset (P < 0.05) were taken forward for confirmation in a second dataset of 72 grade II and III gliomas from northern UK. In this dataset, eight gene variants mapping to five different DNA repair genes (ATM, NEIL1, NEIL2, ERCC6 and RPA4) which were associated with survival. Finally, these eight genetic variants were adjusted for treatment, malignancy grade, patient age and gender, leaving one variant, rs4253079, mapped to ERCC6, with a significant association to survival (OR 0.184, 95% CI 0.054-0.63, P = 0.007). We suggest a possible novel association between rs4253079 and survival in this group of patients with low-grade and anaplastic gliomas that needs confirmation in larger datasets. PMID- 21643988 TI - Preliminary experience of whole-brain radiation therapy (WBRT) in breast cancer patients with brain metastases previously treated with bevacizumab-based chemotherapy. AB - We report our experience of bevacizumab-based chemotherapy (BBCT) followed by whole-brain radiation therapy (WBRT) for breast cancer (BC) patients (pts) with inoperable brain metastases (BM) or who refused surgery. This is a retrospective study of seven metastatic BC pts treated at the Institut Curie with at least one course of BBCT before WBRT, with a delay of <= 12 months between the two treatments. Toxicity was scored according to the common terminology criteria for adverse events (v4. 2010). Median age was 56 years (41-65). Median follow-up was 5.9 months (0.4-24.6). The median dose of bevacizumab was 10 mg/kg. Median number of cycles BBCT was six (5-17). Different chemotherapy regimens were used, the most common combination was paclitaxel-bevacizumab. WBRT was delivered in ten fractions, five fractions/week, for two weeks, to a total of 30 Gy. One pt underwent stereotactic radio surgery (SRS) after WBRT. No pt received BBCT during RT. Most common reported side-effects were nausea (n = 4), headache (n = 3), vomiting (n = 1), and vertigo (n = 3). All pts had mild or moderate grade <= 2 neurologic toxicity. There were no radiological signs of necrosis or cerebral ischemia. BBCT before WBRT was not associated with severe brain toxicity. Because of the limited number of pts, the different BBCT regimens, and important delays between treatments, these results must be confirmed prospectively. PMID- 21643989 TI - Mechanisms of xylanase-induced nitric oxide and phosphatidic acid production in tomato cells. AB - The second messenger nitric oxide (NO), phosphatidic acid (PA) and reactive oxygen species (ROS) are involved in the plant defense response during plant pathogen interactions. NO has been shown to participate in PA production in response to the pathogen-associated molecular pattern xylanase in tomato cell suspensions. Defense responses downstream of PA include ROS production. The goal of this work was to study the signaling mechanisms involved in PA production during the defense responses triggered by xylanase and mediated by NO in the suspension-cultured tomato cells. We analyzed the participation of protein kinases, guanylate cyclase and the NO-mediated posttranslational modification S nitrosylation, by means of pharmacology and biochemistry. We showed that NO, PA and ROS levels are significantly diminished by treatment with the general protein kinase inhibitor staurosporine. This indicates that xylanase-induced protein phosphorylation events might be the important components leading to NO formation, and hence for the downstream regulation of PA and ROS levels. When assayed, a guanylate cyclase inhibitor or a cGMP analog did not alter the PA accumulation. These results suggest that a cGMP-mediated pathway is not involved in xylanase induced PA formation. Finally, the inhibition of protein S-nitrosylation did not affect NO formation but compromised PA and ROS production. Data collectively indicate that upon xylanase perception, cells activate a protein kinase pathway required for NO formation and that, S-nitrosylation-dependent mechanisms are involved in downstream signaling leading to PA and ROS. PMID- 21643990 TI - A pomegranate (Punica granatum L.) WD40-repeat gene is a functional homologue of Arabidopsis TTG1 and is involved in the regulation of anthocyanin biosynthesis during pomegranate fruit development. AB - Anthocyanins are the major pigments responsible for the pomegranate (Punica granatum L.) fruit skin color. The high variability in fruit external color in pomegranate cultivars reflects variations in anthocyanin composition. To identify genes involved in the regulation of anthocyanin biosynthesis pathway in the pomegranate fruit skin we have isolated, expressed and characterized the pomegranate homologue of the Arabidopsis thaliana TRANSPARENT TESTA GLABRA1 (TTG1), encoding a WD40-repeat protein. The TTG1 protein is a regulator of anthocyanins and proanthocyanidins (PAs) biosynthesis in Arabidopsis, and acts by the formation of a transcriptional regulatory complex with two other regulatory proteins: bHLH and MYB. Our results reveal that the pomegranate gene, designated PgWD40, recovered the anthocyanin, PAs, trichome and seed coat mucilage phenotype in Arabidopsis ttg1 mutant. PgWD40 expression and anthocyanin composition in the skin were analyzed during pomegranate fruit development, in two accessions that differ in skin color intensity and timing of appearance. The results indicate high positive correlation between the total cyanidin derivatives quantity (red pigments) and the expression level of PgWD40. Furthermore, strong correlation was found between the steady state levels of PgWD40 transcripts and the transcripts of pomegranate homologues of the structural genes PgDFR and PgLDOX. PgWD40, PgDFR and PgLDOX expression also correlated with the expression of pomegranate homologues of the regulatory genes PgAn1 (bHLH) and PgAn2 (MYB). On the basis of our results we propose that PgWD40 is involved in the regulation of anthocyanin biosynthesis during pomegranate fruit development and that expression of PgWD40, PgAn1 and PgAn2 in the pomegranate fruit skin is required to regulate the expression of downstream structural genes involved in the anthocyanin biosynthesis. PMID- 21643991 TI - Evolutionary significance of an algal gene encoding an [FeFe]-hydrogenase with F domain homology and hydrogenase activity in Chlorella variabilis NC64A. AB - [FeFe]-hydrogenases (HYDA) link the production of molecular H(2) to anaerobic metabolism in many green algae. Similar to Chlamydomonas reinhardtii, Chlorella variabilis NC64A (Trebouxiophyceae, Chlorophyta) exhibits [FeFe]-hydrogenase (HYDA) activity during anoxia. In contrast to C. reinhardtii and other chlorophycean algae, which contain hydrogenases with only the HYDA active site (H cluster), C. variabilis NC64A is the only known green alga containing HYDA genes encoding accessory FeS cluster-binding domains (F-cluster). cDNA sequencing confirmed the presence of F-cluster HYDA1 mRNA transcripts, and identified deviations from the in silico splicing models. We show that HYDA activity in C. variabilis NC64A is coupled to anoxic photosynthetic electron transport (PSII linked, as well as PSII-independent) and dark fermentation. We also show that the in vivo H(2)-photoproduction activity observed is as O(2) sensitive as in C. reinhardtii. The two C. variabilis NC64A HYDA sequences are similar to homologs found in more deeply branching bacteria (Thermotogales), diatoms, and heterotrophic flagellates, suggesting that an F-cluster HYDA is the ancestral enzyme in algae. Phylogenetic analysis indicates that the algal HYDA H-cluster domains are monophyletic, suggesting that they share a common origin, and evolved from a single ancestral F-cluster HYDA. Furthermore, phylogenetic reconstruction indicates that the multiple algal HYDA paralogs are the result of gene duplication events that occurred independently within each algal lineage. Collectively, comparative genomic, physiological, and phylogenetic analyses of the C. variabilis NC64A hydrogenase has provided new insights into the molecular evolution and diversity of algal [FeFe]-hydrogenases. PMID- 21643992 TI - Synergistic effects of Cydia pomonella granulovirus GP37 on the infectivity of nucleopolyhedroviruses and the lethality of Bacillus thuringiensis. AB - To initiate an efficient primary infection, it is important for baculovirus virions to penetrate through the peritrophic membrane (PM) of the host insect. It is frequently reported that enhancins of baculoviruses significantly enhance viral infection by degrading the various protein components of PMs. However, not all baculoviruses encode enhancins. GP37s of baculoviruses share high amino acid identity with fusolins, synergistic factors found in entomopoxviruses. In this study, a truncated Cydia pomonella granulovirus GP37 was expressed in Escherichia coli. The expressed GP37 effectively bound to chitin, and binding occurred predominantly within 3 h. GP37 altered the protein profiles of Spodoptera exigua PMs, from which a 50-kDa protein was dissociated. Droplet-feeding bioassays indicated that GP37 significantly enhanced the infectivity of nucleopolyhedroviruses (NPVs) and the lethality of Bacillus thuringiensis (Bt) in S. exigua larvae. This is the first demonstration of the enhancement of NPVs and Bt infection by a baculovirus GP37. PMID- 21643993 TI - What are the consequences of ant-seed interactions on the abundance of two dry fruited shrubs in a Mediterranean scrub? AB - Strong interactions between dry-fruited shrubs and seed-harvesting ants are expected in early successional scrubs, where both groups have a major presence. We have analysed the implications of the seed characteristics of two dry-fruited shrub species (Coronilla minima and Dorycnium pentaphyllum) on seed predation and dispersal mediated by harvester ants and the consequences of these processes on spatio-temporal patterns of plant abundance in a heterogeneous environment. We found that large C. minima seeds were collected much more (39%) than small D. pentaphyllum seeds (2%). However, not all of the removed seeds of these plant species were consumed, and 12.8% of the seeds were lost along the trails, which increased dispersal distances compared with abiotic dispersal alone. Seed dropping occurred among all microhabitats of the two plant species, but especially in open microhabitats, which are the most suitable ones for plant establishment. The two plant species increased their presence in the study area during the study period: C. minima in open microhabitats and D. pentaphyllum in high vegetation. The large size of C. minima seeds probably limited the primary seed dispersal of this species, but may have allowed strong interaction with ants. Thus, seed dispersal by ants resulted in C. minima seeds reaching more suitable microhabitats by means of increasing dispersal distance and redistribution among microhabitats. In contrast, the smaller size of D. pentaphyllum seeds arguably allows abiotic seed dispersal over longer distances and colonization of all types of microhabitats, although it probably also limits their interaction with ants and, consequently, their redistribution in suitable microhabitats. We suggest that dyszoochory could contribute to the success of plant species with different seed characteristics in scrub habitats where seeds are abundantly collected by seed-harvesting ants. PMID- 21643994 TI - The island syndrome and population dynamics of introduced rats. AB - The island syndrome predicts directional changes in the morphology and demography of insular vertebrates, due to changes in trophic complexity and migration rates caused by island size and isolation. However, the high rate of human-mediated species introductions to some islands also increases trophic complexity, and this will reduce the perceived insularity on any such island. We test four hypotheses on the role of increased trophic complexity on the island syndrome, using introduced black rats (Rattus rattus) on two isolated coral atolls in the Mozambique Channel. Europa Island has remained relatively pristine and insular, with few species introductions, whereas Juan de Nova Island has had many species introductions, including predators and competitors of rats, anthropogenically increasing its trophic complexity. In the most insular environments, the island syndrome is expected to generate increases in body size and densities of rodents but decreases in the rates of reproduction and population cycling. Morphology and reproduction were compared using linear regression and canonical discriminant analysis, while density and population cycling were compared using spatially explicit capture-recapture analysis. Results were compared to other insular black rat populations in the Mozambique Channel and were consistent with predictions from the island syndrome. The manifestation of an island syndrome in rodents depends upon the trophic composition of a community, and may not relate to island size alone when many species additions, such as invasions, have occurred. The differing patterns of rodent population dynamics on each island provide information for future rodent eradication operations. PMID- 21643995 TI - Fire management to combat disease: turning interactions between threats into conservation management. AB - As the number and intensity of threats to biodiversity increase, there is a critical need to investigate interactions between threats and manage populations accordingly. We ask whether it is possible to reduce the effects of one threat by mitigating another. We used long-term data for the long-lived resprouter, Xanthorrhoea resinosa Pers., to parameterise an individual-based population model. This plant is currently threatened by adverse fire regimes and the pathogen Phytophthora cinnamomi. We tested a range of fire and disease scenarios over various time horizons relevant to the population dynamics of the species and the practicalities of management. While fire does not kill the disease, it does trigger plant demographic responses that may promote population persistence when disease is present. Population decline is reduced with frequent fires because they promote the greatest number of germination events, but frequent fires reduce adult stages, which is detrimental in the long term. Fire suppression is the best action for the non-seedling stages but does not promote recruitment. With disease, frequent fire produced the highest total population sizes for shorter durations, but for longer durations fire suppression gave the highest population sizes. When seedlings were excluded, fire suppression was the best action. We conclude that fire management can play an important role in mitigating threats posed by this disease. The best approach to reducing declines may be to manage populations across a spatial mosaic in which the sequence of frequent fires and suppression are staggered across patches depending on the level of disease at the site. PMID- 21643996 TI - Disturbance governs dominance of an invasive forb in a temporary wetland. AB - Dominance of invasive species is often assumed to be due to a superior ability to acquire resources. However, dominance in plant communities can arise through multiple interacting mechanisms, including disturbance. Inter-specific competition can be strongly affected by abiotic conditions, which can determine the outcome of competitive interactions. We evaluated competition and disturbance as mechanisms governing dominance of Phyla canescens (hereafter lippia), an invasive perennial forb from South America, in Paspalum distichum (perennial grass, hereafter water couch) meadows in floodplain wetlands of eastern Australia. Water couch meadows (in the study area) are listed under the Ramsar Convention due to their significance as habitat for migratory waterbirds. In the field, we monitored patterns of vegetation boundaries between the two species in response to flooding. Under controlled glasshouse conditions, we explored competitive interactions between the native water couch and lippia subject to different soil moisture/inundation regimes. We did this using a pairwise factorial glasshouse experiment that manipulated neighbor density (9 treatments) and soil moisture/inundation (4 treatments). In the field trial, inundation increased the cover of water couch. Under more controlled conditions, the invader had a competitive effect on the native species only under dry soil conditions, and was strongly inhibited by inundation. This suggests that dry conditions favor the growth of the invader and wetter (more historical) conditions favor the native grass. In this system, invader dominance is governed by altered disturbance regimes which give the invader a competitive advantage over the native species. PMID- 21643997 TI - The protective effects of inosine against chemical hypoxia on cultured rat oligodendrocytes. AB - Inosine is a purine nucleoside and is considered protective to neural cells including neurons and astrocytes against hypoxic injury. However, whether oligodendrocytes (OLs) could also be protected from hypoxia by inosine is not known. Here we investigated the effects of inosine on primarily cultured rat OLs injured by rotenone-mediated chemical hypoxia, and the mechanisms of the effects using ATP assay, MTT assay, PI-Hoechst staining, TUNEL, and immunocytochemistry. Results showed that rotenone exposure for 24 h caused cell death and impaired viability in both immature and mature OLs, while pretreatment of 10 mM inosine 30 min before rotenone administration significantly reduced cell death and improved the viability of OLs. The same concentration of inosine given 120 min after rotenone exposure also improved viability of injured mature OLs. Immunocytochemistry for nitrotyrosine and cellular ATP content examination indicated that inosine may protect OLs by providing ATP and scavenging peroxynitrite for cells. In addition, immature OLs were more susceptible to hypoxia than mature OLs; and at the similar degree of injury, inosine protected immature and mature OLs differently. Quantitative real-time PCR revealed that expression of adenosine receptors was different between these two stages of OLs. These data suggest that inosine protect OLs from hypoxic injury as an antioxidant and ATP provider, and the protective effects of inosine on OLs vary with cell differentiation, possibly due to the adenosine receptors expression profile. As OLs form myelin in the central nervous system, inosine could be used as a promising drug to treat demyelination-involved disorders. PMID- 21643998 TI - Effect of acupuncture on hypothalamic-pituitary-adrenal system in maternal separation rats. AB - The maternal separation (MS) animal model has been widely used to study early life stress and several psychiatric conditions such as depression and anxiety. In this study, we investigated the effect of acupuncture on anxiety-related behaviors and hypothalamic-pituitary-adrenal (HPA) system in MS-induced early life stress of Sprague-Dawley rat pups (14-21 postnatal days). For determining anxiety-related behaviors, the elevated plus-maze test was performed. The effects of acupuncture on the activation of stress were measured by assessing plasma levels of corticosterone (CORT) and adrenocorticotropin hormone (ACTH). The hypothalamic immunoreactivity (IR) of arginine vasopressin (AVP) was also examined. Acupuncture was conducted at acupoint HT7, which is used to treat mental disorders in Oriental medicine, for seven consecutive days. Acupuncture significantly decreased the frequencies of open arm entries and the amount of time spent in the open arms in MS rats. In addition, acupuncture reduced CORT and ACTH levels in plasma of MS rats, and AVP-IR in the hypothalamic paraventricular nucleus of MS rats. In conclusion, acupuncture reduced anxiety-related behaviors and modulated the HPA system. These findings suggest that acupuncture at HT7 may be useful as a therapeutic treatment in MS-induced early life stress. PMID- 21643999 TI - Expression of pancreatitis-associated protein after traumatic brain injury: a mechanism potentially contributing to neuroprotection in human brain. AB - Neuronal cell death after severe traumatic brain injury (TBI) is caused by a complex interplay of pathological mechanisms including excitotoxicity, oxidative stress, mitochondrial dysfunction, extensive neuroinflammation, and ischemia reperfusion injury. Pancreatitis-associated protein I (PAP I/reg2) was reported to be a survival factor for peripheral neurons, particularly sensory and motor neurons. In rat brains, by experimental TBI as well as by kainic acid induced brain seizure, PAP I and PAP III were found to be up-regulated in central neurons. In this study, we performed immunohistochemical staining in postmortem human brain from patients who died after severe TBI to demonstrate PAP expression on protein level in cerebellar Purkinje cells, pyramidal and granular neurons in cerebral cortex, and cortical neurons in the fore- and mid-brain. In primary cultures of rat brain cortical, hippocampal, and cerebellar neurons, we found neuroprotective effects for PAP I on H(2)O(2)-induced oxidative stress. Moreover, serum K(+)-deprivation induces apoptotic cell death in 55% of cerebellar granule neurons (CGN), whereas upon treatment with PAP I only 32% of CGN are apoptotic. Using Western blot analyses, we compared protein phosphorylation in neuronal signaling pathways activated by PAP I versus Interleukin-6 (IL-6). We found a rapid activation of Akt-kinase phosphorylation by PAP I with a peak at 15 min, whereas IL-6 induces Akt-phosphorylation lasting longer than 30 min. Phosphorylation of MAP-42/44 kinases is stimulated in a comparable fashion. Both, IL-6 and PAP I increase phosphorylation of NFkappaB for activation of gene transcription, whereas only IL-6 recruits STAT3 phosphorylation, indicating that STAT3 is not a target of PAP I transcription activation in brain neurons. Application of the Akt-inhibitor Wortmanin reveals only a partial inhibition of PAP I-dependent protection of CGN from H(2)O(2)-induced oxidative stress. Based on our findings, we suggest that PAP I is a long lasting neurotrophic signal for central neurons. The neuroprotective effects parallel those that have been described for effects of PAP I in ciliary neurotrophic factor (CNTF)-mediated survival of sensory and motor neurons. PAP I may act in autocrine and/or paracrine fashion and thus may contribute to endogenous protective mechanisms relevant under harmful conditions like oxidative stress, brain injury, or neurodegeneration. PMID- 21644000 TI - The pro-apoptotic BH3-only protein Bid is dispensable for development of insulitis and diabetes in the non-obese diabetic mouse. AB - Type 1 diabetes is caused by death of insulin-producing pancreatic beta cells. Beta-cell apoptosis induced by FasL may be important in type 1 diabetes in humans and in the non-obese diabetic (NOD) mouse model. Deficiency of the pro-apoptotic BH3-only molecule Bid protects beta cells from FasL-induced apoptosis in vitro. We aimed to test the requirement for Bid, and the significance of Bid-dependent FasL-induced beta-cell apoptosis in type 1 diabetes. We backcrossed Bid-deficient mice, produced by homologous recombination and thus without transgene overexpression, onto a NOD genetic background. Genome-wide single nucleotide polymorphism analysis demonstrated that diabetes-related genetic regions were NOD genotype. Transferred beta cell antigen-specific CD8+ T cells proliferated normally in the pancreatic lymph nodes of Bid-deficient mice. Moreover, Bid deficient NOD mice developed type 1 diabetes and insulitis similarly to wild-type NOD mice. Our data indicate that beta-cell apoptosis in type 1 diabetes can proceed without Fas-induced killing mediated by the BH3-only protein Bid. PMID- 21644001 TI - Behavior, intention or chance? A longitudinal study of HIV seroadaptive behaviors, abstinence and condom use. AB - Seroadaptive behaviors have been widely described as preventive strategies among men who have sex with men (MSM) and other populations worldwide. However, causal links between intentions to adopt seroadaptive behaviors and subsequent behavior have not been established. We conducted a longitudinal study of 732 MSM in San Francisco to assess consistency and adherence to multiple seroadaptive behaviors, abstinence and condom use, whether prior intentions predict future seroadaptive behaviors and the likelihood that observed behavioral patterns are the result of chance. Pure serosorting (i.e., having only HIV-negative partners) among HIV negative MSM and seropositioning (i.e., assuming the receptive position during unprotected anal sex) among HIV-positive MSM were more common, more successfully adhered to and more strongly associated with prior intentions than consistent condom use. Seroconcordant partnerships occurred significantly more often than expected by chance, reducing the prevalence of serodiscordant partnerships. Having no sex was intended by the fewest MSM, yet half of HIV-positive MSM who abstained from sex at baseline also did so at 12 month follow-up. Nonetheless, no preventive strategy was consistently used by more than one-third of MSM overall and none was adhered to by more than half from baseline to follow-up. The effectiveness of seroadaptive strategies should be improved and used as efficacy endpoints in trials of behavioral prevention interventions. PMID- 21644002 TI - Female parthenogenetic apomixis and androsporogenesis in Douglas-fir embryonal initials in an artificial sporangium. AB - Control of female parthenogenetic apomixis and androsporogenesis of Douglas-fir embryonal initials was studied using an experimental culture system in which changes in growth condition can mediate changes in cell identity and outcomes. This culture system constitutes an artificial sporangium in which myriad culture conditions can be simulated and should be applicable for research on a variety of gymnosperms. In this study, embryonal initials from developing seeds from two Douglas-fir trees were rescued and became reprogrammed for female parthenogenetic apomixis (fPA) and parthenogenetic androsporogenesis (mPA). Female PA was initiated by endomitosis forming a binucleate cell with a diploid egg-equivalent and an apoptotic ventral canal nucleus in an archegonial tube. Egg-equivalent nuclei formed cells (parthenotes) that were discharged into an aqueous culture medium. Parthenotes developed axial tiers atypical of early embryogenesis in seeds. Earlier in the year, androsporangial tubes were parthenogenetically differentiated and released monads, dyads, triads, and tetrads into the culture medium. Spores showed chromosomal aberrations. PA demonstrated a temporal separation in gender expression (dichogamy). Embryonal initials brought forward and by-passed the long juvenile phases normally needed for cells to develop into trees and express reproductive maturity. Expressions of fPA and mPA indicated that the specialized culture flasks served as an artificial sporangium (AS). Awareness is raised for the value of an AS for research in gymnosperm life cycles and as a teaching and research laboratory. PMID- 21644003 TI - New data about the suspensor of succulent angiosperms: Ultrastructure and cytochemical study of the embryo-suspensor of Sempervivum arachnoideum L. and Jovibarba sobolifera (Sims) Opiz. AB - The development of the suspensor in two species - Sempervivum arachnoideum and Jovibarba sobolifera - was investigated using cytochemical methods, light and electron microscopy. Cytological processes of differentiation in the embryo suspensor were compared with the development of embryo-proper. The mature differentiated suspensor consists of a large basal cell and three to four chalazal cells. The basal cell produces haustorial branched invading ovular tissues. The walls of the haustorium and the micropylar part of the basal cell form the wall ingrowths typical for a transfer cells. The ingrowths also partially cover the lateral wall and the chalazal wall separating the basal cell from the other embryo cells. The dense cytoplasm filling the basal cell is rich in: numerous polysomes lying free or covering rough endoplasmic reticulum (RER), active dictyosomes, microtubules, bundles of microfilaments, microbodies, mitochondria, plastids and lipid droplets. Cytochemical tests (including proteins, insoluble polysaccharides and lipids are distributed in the suspensor during different stages of embryo development) showed the presence of high amounts of macromolecules in the suspensor cells, particularly during the globular and heart-shaped phases of embryo development. The protein bodies and lipid droplets are the main storage products in the cells of the embryo-proper. The results of Auramine 0 indicate that a cuticular material is present only on the surface walls of the embryo-proper, but is absent from the suspensor cell wall. The ultrastructural features and cytochemical tests indicate that in the two species - S. arachnoideum and J. sobolifera - the embryo-suspensor is mainly involved in the absorption and transport of metabolites from the ovular tissues to the developing embryo-proper. PMID- 21644004 TI - Application of loop-mediated isothermal amplification (LAMP)-based technology for authentication of Catharanthus roseus (L.) G. Don. AB - In this study, loop-mediated isothermal amplification (LAMP)-based molecular marker was developed for authentication of Catharanthus roseus, a medicinal plant. Samples of this plant were collected from different geographical locations in India. Random amplified polymorphic deoxyribonucleic acid (DNA) analysis of collected samples was carried out with 25 random primers. A 610-bp DNA fragment, common to all accessions, was eluted, cloned, and sequenced. Four LAMP primers were designed on the basis of sequence of 610 bp DNA fragment. LAMP reaction, containing 10* Bst DNA polymerase reaction buffer, Bst DNA polymerase, four in house designed primers, dNTPs, MgSO(4), and betaine, was incubated at 65 degrees C for 1 h. The resulting amplicon was visualized by adding SYBR Green I to the reaction tube. The data showed confirmatory results. Since the assay method is simple, sensitive, and cost-effective, it is a feasible method for identifying and authentication of C. roseus. PMID- 21644005 TI - Teaching rural women in Nicaragua the principles of breast health. AB - Breast cancer has emerged as an important health condition worldwide, including developing countries. Screening is limited or non-existent in resource-poor areas. The purpose of this study was to assess knowledge, attitudes, and practices of self (SBE) and clinical (CBE) breast examinations among 198 rural Nicaraguan women. Ten (5.1%) had performed a SBE, and 16 (8.1%) had a CBE. CBE was significantly associated with a pre-instruction total score of 70% or greater (OR = 13.7, 95% CI = 1.26, 149.70, p = 0.03). Family history of breast cancer was significantly associated with performing a SBE (OR = 5.5, 95% CI = 1.10, 27.81, p = 0.037) and a CBE (OR = 7.1, 95% CI = 1.40, 35.94, p = 0.018). A CBE is a significant determinant of pre-existing breast health knowledge. Physicians or physician extenders should be encouraged to perform a CBE as a routine component of health care delivery and cancer screening for women. PMID- 21644006 TI - The International Core Literature Consensus (ICLC): an alternative curriculum for Oncologists. AB - Oncologists must familiarize themselves with a complex evidence base. Several curricula have been devised, but it is not clear how these are used. We chose breast cancer, since this has a large literature base. Of the 285 radiation and medical oncology trainees in the USA and UK responding to our survey, over 90% reported knowledge of the literature was essential. Just over half of respondents had actually read the ASCO or ESMO curricula, with only 23% reporting that the curricula were important in their learning. Our survey revealed dissatisfaction with current curricula and a demand for more pragmatic literature guidance. We designed an alternative curriculum by using a 21-member peer review group to validate a list of key papers in breast oncology. Oncologists in training need guidance to direct their study. A curriculum based on an International Core Literature Consensus might match the needs of trainees more closely. PMID- 21644007 TI - Assessing the effect of darbepoetin alfa on patient-reported fatigue in chemotherapy-induced anemia in four randomized, placebo-controlled clinical trials. AB - PURPOSE: This analysis examined the effects of darbepoetin alfa on hemoglobin and fatigue outcomes in patients with cancer using latent growth curve modeling (LGM). METHODS: Data from 4 clinical trials of darbepoetin alfa in lung cancer (2 studies; n = 547; n = 288), lymphoproliferative malignancies (n = 339), and non myeloid malignancies (n = 320) were analyzed separately. Fatigue was assessed using the FACT-Fatigue (FACT-F) scale. Effects of darbepoetin alfa on changes in hemoglobin and FACT-F scores were evaluated using LGM, controlling for age, gender, Eastern Cooperative Oncology Group performance status, health status, and total transfusions. RESULTS: Patients receiving darbepoetin alfa had higher rates of change in hemoglobin (standardized regression coefficient [[Formula: see text]] = 0.30 to 0.53, all P < 0.05) than placebo. Patients with greater rates of change in hemoglobin reported improvements in fatigue outcomes ([Formula: see text] = 0.28 to 0.59, all P < 0.05). The total standardized effect of darbepoetin alfa on fatigue outcomes corresponded to a mean change of 0.9 to 3.5 points in FACT-F scores, with one trial demonstrating changes exceeding the minimal important difference of 3 points. CONCLUSIONS: Darbepoetin alfa improved hemoglobin which was associated with improved fatigue across the 4 trials. Clinically, meaningful improvement in fatigue was seen in 2 trials. More complex statistical analysis models of treatment may assist in understanding the effects of erythropoiesis-stimulating agents on patient-reported outcomes. PMID- 21644008 TI - Comparison of the effects of different electrocautery applications to peripheral nerves: an experimental study. AB - BACKGROUND: This study was designed to investigate the effects of bipolar and mononopolar electrocauterization on peripheral nerve tissue. The comparison on the deleterious effects of the different cautery modalities and the importance of probe tip placement are evaluated using electrophysiological, electron microscopic and biochemical assessment parameters. METHODS: Ninety-eight male Wistar albino rats, each weighing 250-275 g, were randomly divided into 14 groups. Each group consisted of seven animals. Monopolar and bipolar electrocautery were performed at 15 watts. The application was performed either directly on the nerve or 1 mm lateral to the longitudinal axis of the nerve for 'near the nerve groups', respectively. RESULTS: The electrophysiological findings showed that the mean amplitudes were at the lowest value in the first day for all the groups. At the end of the 3rd week, we recognised that the electrophysiological recovery continued. Electron microscopic evaluation showed myelin disruption in all groups. Myelin disruption of healthy neurons was at the highest level in the 1st day of application in accordance with the electrophysiological findings. Biochemical evaluation revealed statistical significance between the control and the two of the 'near the nerve groups' (GIII and GV) for NO (nitrite and nitrate) serum level. CONCLUSION: The data of the present study might suggest that electrocautery, independent of the type and form of application, may result in significant damage in histological and electrophysological basis. Although the relative proportions cannot be ascertained, the time course of recovery suggests that both axon and myelin damage have occurred. The probable electrocautery damage may be of substantial importance for the situation that the nerves are displaced by tumor masses or atypical neural traces. PMID- 21644009 TI - Genetic diversity of 'Uba' mango tree using ISSR markers. AB - In this study, the genetic diversity of 'Uba' mango trees cultivated at the Zona da Mata of Minas Gerais State, Brazil, was assessed, to identify whether there is variability in the plants grown in the region, justifying the mass selection as a breeding method. We used 102 accessions. Leaves were collected for extraction of genomic DNA, which was amplified with nine ISSR primers. The data obtained by the analysis of electrophoretic patterns were arranged in a binary matrix, considering 0 for the absence and 1 for the presence of bands. Based on these data, we performed the analysis of genetic dissimilarity and carried out the cluster analysis by the methods of Tocher and graphical dispersion. The most similar accessions are 144 and 150, both coming from Uba, while the most divergent ones are 29 and 97, from Visconde do Rio Branco. The grouping by the Tocher method separated the accessions into six groups, 94.1% of which were allocated in the first group and showed that there is no separation of accessions depending on the sampling sites. The 3D scatter plot reinforces this conclusion. There is genetic variability among the accessions of 'Uba' mango tree evaluated. Therefore, it is possible to make mass selection in open-pollinated populations. PMID- 21644010 TI - Glycated albumin but not HbA1c reflects glycaemic control in patients with neonatal diabetes mellitus. AB - AIMS/HYPOTHESIS: It is difficult to use HbA(1c) as an indicator of glycaemic control in patients with neonatal diabetes mellitus (NDM) because of high levels of fetal haemoglobin (HbF) remaining in the blood. In this study, glycated albumin (GA), which is not affected by HbF, and HbA(1c) were compared to evaluate whether they reflect glycaemic control in patients with NDM. METHODS: This study included five patients with NDM. Age at diagnosis was 38 +/- 20 days. Insulin therapy was started in all patients, and levels of GA, HbA(1c) and HbF were measured monthly for 6 months. One-month average preprandial plasma glucose (aPPG) was calculated using self-monitoring of blood glucose. RESULTS: Plasma glucose and GA were elevated (29.7 +/- 13.1 mmol/l [n = 5] and 33.3 +/- 6.9% [n = 3], respectively) but HbA(1c) was within normal limits (5.4 +/- 2.6% [35.5 +/- 4.9 mmol/mol]; n = 4) at diagnosis. With diabetes treatment, aPPG (r = -0.565, p = 0.002), GA (r = -0.552, p = 0.003) and HbF (r = -0.855, p < 0.0001) decreased with age, whereas HbA(1c) increased (r = 0.449, p = 0.004). GA was strongly positively correlated with aPPG (r = 0.784, p < 0.0001), while HbA(1c) showed no correlation with aPPG (r = 0.221, p = 0.257) and was significantly inversely correlated with HbF (r = -0.539, p = 0.004). CONCLUSIONS/INTERPRETATION: GA is a useful indicator of glycaemic control in patients with NDM, whereas HbA(1c) is influenced by age-related changes in HbF and does not accurately reflect glycaemic control. PMID- 21644011 TI - Study of the pharmacokinetic and pharmacogenetic contribution to the toxicity of high-dose methotrexate in children with acute lymphoblastic leukemia. AB - Methotrexate inhibits the conversion of 5,10-methylenetetrahydrofolate to 5 methyltetrahydrofolate by methylenetetrahydrofolate reductase (MTHFR). MTHFR has a common functional polymorphism C677T. The present study aimed to investigate the prevalence of MTHFR polymorphisms in Egyptian children with ALL and the relation to MTX-related toxicity, relapse, and MTX pharmacokinetic parameters. Forty patients with ALL were included in the study. They were treated according to ALL-NCI total XIII protocol. MTX-related toxicity and MTX pharmacokinetic parameters were assessed during therapy. MTHFR genotyping was done with a PCR based restriction fragment length polymorphism assay, and MTX pharmacokinetic parameters were assessed by HPLC. The MTHFR C677T polymeric allele frequencies were 55, 35, and 10% for CC, CT, and TT genotypes, respectively, among the studied patients with ALL. MTX therapy was significantly associated with toxicity signs in TT genotype: elevated transaminases (P < 0.0001), elevated serum alpha 1 microglobulin protein (P < 0.0001), anemia (P < 0.0001), neutropenia (P < 0.0001), thrombocytopenia (P < 0.0001), and elevated CSF-beta-glucuronidase activity (P < 0.0001). Patients with TT genotype showed significant increase in MTX t(1/2) and AUC (P < 0.0001), while MTX elimination rate and total body clearance were significantly decreased (P < 0.0001 and P < 0.05, respectively) compared with CC genotype. The TT genotype was significantly associated with relapse in 2 years in 50% compared with 28.57% in CT and 13.64% in CC alleles. The overall 2-year survival was significantly lower in TT genotype (50%) compared with CC genotype (90.91%; P = 0.01). MTHFR TT genotype is significantly associated with increased toxicity during methotrexate therapy as well as increased relapse rate in pediatric patients with ALL. In future, MTX dose adjustment in ALL treatment protocols should be considered based on patient's genotype. PMID- 21644012 TI - Extrapulmonary small cell: a novel case of small cell carcinoma of the thyroid gland. AB - Neuroendocrine tumors comprise a large group of malignancies which share unique morphological features and are characterized by the presence of neuroendocrine markers such as synaptophysin, chromogranin-A, and CD56 (N-CAM), ranging from indolent tumors, such as carcinoid tumors, to aggressive tumors, such as small cell carcinoma. The lung is the most common site for primary neuroendocrine tumors. Extrapulmonary primary sites of small cell carcinoma are rare but have been documented arising from various sites including esophagus, stomach, colon and rectum, gallbladder, thymus, salivary gland, ovary, cervix, bladder, prostate, and skin. We present a case of small cell carcinoma arising from the thyroid gland, a site not previously described in the literature. A 59-year-old woman presented with a thyroid mass, which, after resection, showed small cell morphology and positive immunostains for TTF-1, synaptophysin, chromogranin-A, CD56, etc. Five months after diagnosis, she had widely metastatic disease. After a near-complete response to the first chemo-treatment, her disease progressed. Following local radiation and more rounds of chemotherapy, she succumbed to the disease, 15 months after diagnosis. Our patient had no pulmonary lesions at the time of diagnosis to suggest metastasis from the lung. Much like its pulmonary counterparts, this small cell carcinoma of primary thyroid origin displayed an aggressive clinical course and poor outcome. Although it shows early sensitivity to chemotherapy, small cell carcinoma remains a difficult-to-treat cancer with a poor prognosis and can rarely be seen originating in organs outside of the lung. PMID- 21644013 TI - Hypoplastic left heart syndrome and aortic atresia-mitral stenosis variant: role of myocardial protection strategy and impact of ventriculo-coronary connections after stage I palliation. AB - Aortic atresia-mitral stenosis (AA-MS) has been implicated as a determinant of outcome after Stage-1 palliation (S1P) in hypoplastic left heart syndrome (HLHS).Studies evaluating the association of AA-MS with ventriculo-coronary connections (VCC) and mortality report conflicting results. The significance of VCC, myocardial protection, and shunt strategy after S1P has yet to be determined. Between January 2005 and July 2009, 100 neonates with HLHS underwent S1P. Mitral and aortic valves and presence of VCC were assessed. Antegrade continuous cold blood cardioplegia was administered throughout the vast extent of the neo-aortic reconstruction. A right ventricle-to-pulmonary shunt was used for an ascending aortic diameter of 0.6 mm/kg or less. Survival analysis was performed to determine predictors and assess impact of AA-MS and VCC on hospital and interstage mortality. Twenty-seven (of 100) patients had AA-MS. The mean age and weight at S1P were 6.5 +/- 2.8 days and 3.09 +/- 0.47 kg, respectively. VCC were found in 56% of AA-MS. Twenty-two had Norwood-Sano, 3 had classic Norwood, and 2 had hybrid S1P. VCC were associated with AA-MS, endocardial fibroelastosis, and ascending aortic size <2 mm (P < 0.05) but not higher mortality (P = ns). Operative and interstage survival for AA-MS after S1P was 85.2 and 71%, respectively (not statistically different compared to all other subtypes; P = ns). Actuarial survival after S1P at 1, 3, 6, 12, and 36 months was 92.9 +/- 4.9, 78.6 +/- 7.8, 75 +/- 8.2, 71.3 +/- 8.3, and 71.3 +/- 8.3%, respectively. Intact atrial septum and post-S1P renal dysfunction (P < 0.05) were independent predictors of hospital and interstage mortality. In patients with HLHS, AA-MS carries no survival disadvantage after S1P during the hospital and interstage period regardless of VCC. Intact atrial septum and post-S1P renal dysfunction predict early and interstage mortality. Myocardial protection and shunt strategy might influence the outcome in this HLHS variant. PMID- 21644014 TI - Molecular modeling studies on phosphonic acid-containing thiazole derivatives: design for fructose-1,6-bisphosphatase inhibitors. AB - Presently, an in silico modeling was carried out on a series of 63 phosphonic acid-containing thiazole derivatives as fructose-1,6-bisphosphatase (FBPase) inhibitors using CoMFA/CoMSIA and molecular docking methods. The CoMFA and CoMSIA models using 51 molecules in the training set gave r (cv) (2) values of 0.675 and 0.619, r ( 2 ) values of 0.985 and 0.979, respectively. The systemic external validation indicated that our CoMFA and CoMSIA models possessed high predictive powers with r (0) (2) values of 0.995 and 0.994, r (m(test)) (2) values of 0.887 and 0.860, respectively. The 3D contour maps of the CoMFA and CoMSIA provided smooth and interpretable explanation of the structure-activity relationship for the inhibitors. Molecular docking studies revealed that a phosphonic group was essential for binding to the AMP binding site, and some key features were also identified. The analyses of the 3D contour plots and molecular docking results permitted interesting conclusions about the effects of different substituent groups at different positions of the common scaffold, which might guide the design of novel FBPase inhibitors with higher activity and bioavailability. A set of 60 new analogues were designed by utilizing the results revealed in the present study, and were predicted with significantly improved potencies in the developed models. The findings can be quite useful to aid the designing of new fructose-1,6-biphophatase inhibitors with improved biological response. PMID- 21644015 TI - Dispersal constraints for stream invertebrates: setting realistic timescales for biodiversity restoration. AB - Biodiversity goals are becoming increasingly important in stream restoration. Typical models of stream restoration are based on the assumption that if habitat is restored then species will return and ecological processes will re-establish. However, a range of constraints at different scales can affect restoration success. Much of the research in stream restoration ecology has focused on habitat constraints, namely the in-stream and riparian conditions required to restore biota. Dispersal constraints are also integral to determining the timescales, trajectory and potential endpoints of a restored ecosystem. Dispersal is both a means of organism recolonization of restored sites and a vital ecological process that maintains viable populations. We review knowledge of dispersal pathways and explore the factors influencing stream invertebrate dispersal. From empirical and modeling studies of restoration in warm-temperate zones of New Zealand, we make predictions about the timescales of stream ecological restoration under differing levels of dispersal constraints. This process of constraints identification and timescale prediction is proposed as a practical step for resource managers to prioritize and appropriately monitor restoration sites and highlights that in some instances, natural recolonization and achievement of biodiversity goals may not occur. PMID- 21644016 TI - Understanding and integrating local perceptions of trees and forests into incentives for sustainable landscape management. AB - We examine five forested landscapes in Africa (Cameroon, Madagascar, and Tanzania) and Asia (Indonesia and Laos) at different stages of landscape change. In all five areas, forest cover (outside of protected areas) continues to decrease despite local people's recognition of the importance of forest products and services. After forest conversion, agroforestry systems and fallows provide multiple functions and valued products, and retain significant biodiversity. But there are indications that such land use is transitory, with gradual simplification and loss of complex agroforests and fallows as land use becomes increasingly individualistic and profit driven. In Indonesia and Tanzania, farmers favor monocultures (rubber and oil palm, and sugarcane, respectively) for their high financial returns, with these systems replacing existing complex agroforests. In the study sites in Madagascar and Laos, investments in agroforests and new crops remain rare, despite government attempts to eradicate swidden systems and their multifunctional fallows. We discuss approaches to assessing local values related to landscape cover and associated goods and services. We highlight discrepancies between individual and collective responses in characterizing land use tendencies, and discuss the effects of accessibility on land management. We conclude that a combination of social, economic, and spatially explicit assessment methods is necessary to inform land use planning. Furthermore, any efforts to modify current trends will require clear incentives, such as through carbon finance. We speculate on the nature of such incentive schemes and the possibility of rewarding the provision of ecosystem services at a landscape scale and in a socially equitable manner. PMID- 21644017 TI - Characterization and application of a diamine oxidase from Lathyrus sativus as component of an electrochemical biosensor for the determination of biogenic amines in wine and beer. AB - In this work, we have characterized a diamine oxidase (DAO) from Lathyrus sativus and evaluated its use, for the first time, as biocatalytic component of an electrochemical biosensor for the determination of biogenic amines index in wine and beer samples. Firstly, DAO was electrokinetically characterized free in solution by means of a platinum electrode and then immobilized by using polyazetidine prepolimer on the surface of screen-printed electrodes constituted of two gold working electrodes. The amperometric measurements were carried out by using a flow system at a fixed potential of +600 mV vs the internal silver pseudo reference in phosphate buffer solution (0.1 mol l(-1), pH = 7.4). The analysis of wine and beer samples were performed in flow injection system using the dual channel transducer providing simultaneous detection of sample and blank signal, and the resulting signal (after subtraction of the blank signal) was referred to that of putrescine. The results were compared with those obtained using a modified reference method based on gas chromatography-mass spectrometry analysis on the same samples. The results obtained in the analysis of Italian wines shows the better suitability of DAO-based biosensor in the determination of the biogenic amines (BAs) index expressed as putrescine equivalent in both red and white wines, being less efficient in beer samples where it underestimates by about 50% the BAs content. PMID- 21644018 TI - Screening nucleotide binding to amino acid-coated supports by surface plasmon resonance and nuclear magnetic resonance. AB - Here, we describe a rapid and efficient screening method using surface plasmon resonance (SPR) and saturation transfer difference-nuclear magnetic resonance (STD-NMR) spectroscopy to yield information regarding the residues involved in nucleotide binding to amino acid-coated supports. The aim of this work was to explore the use of these spectroscopic techniques to study amino acid-nucleotide interactions in order to improve the binding specificity of the amino acid ligands used to purify plasmid DNA. For SPR, we present a strategy that immobilizes arginine and lysine on a surface as model supports, and we analyze binding responses when synthetic homo-deoxyoligonucleotides are injected over the amino acid surface. The binding responses are detectable and reproducible despite the small size of the immobilized amino acids. Using STD-NMR, we performed epitope mapping of homo-deoxyoligonucleotides bound to L-arginine-bisoxyran Sepharose and L-lysine-Sepharose supports. Polynucleotide binding preferences differed; for example, polyC interacted preferentially through its backbone with the two supports, whereas polyT bound the supports through its thymine moiety. STD-NMR combined with SPR measurements was successfully used to screen amino acid nucleotide interactions and determine the binding affinities of the complexes. PMID- 21644019 TI - Gold nanomaterials as a new tool for bioanalytical applications of laser desorption ionization mass spectrometry. AB - Nanomaterials have emerging importance in laser desorption ionization mass spectrometry (LDI-MS) with the ultimate objective being to overcome some of the most important limitations intrinsically related to the use of conventional organic matrices in matrix-assisted (MA) LDI-MS. This review provides a critical overview of the most recent literature on the use of gold nanomaterials as non conventional desorption ionization promoters in LDI-MS, with particular emphasis on bioanalytical applications. Old seminal papers will also be discussed to provide a timeline of the most significant achievements in the field. Future prospects and research needs are also briefly discussed. PMID- 21644021 TI - Availability of tobacco and alcohol products in Los Angeles community pharmacies. AB - The availability of tobacco and alcohol products in community pharmacies contradicts the pharmacists' Code of Ethics and presents challenges for a profession that is overwhelmingly not in favor of the sale of these products in its practice settings. The primary aim of this study was to estimate the proportion of pharmacies that sell tobacco products and/or alcoholic beverages and to characterize promotion of these products. The proportion of pharmacies that sell non-prescription nicotine replacement therapy (NRT) products as aids to smoking cessation also was estimated. Among 250 randomly-selected community pharmacies in Los Angeles, 32.8% sold cigarettes, and 26.0% sold alcohol products. Cigarettes were more likely to be available in traditional chain pharmacies and grocery stores than in independently-owned pharmacies (100% versus 10.8%; P < 0.001), and traditional chain drug stores and grocery stores were more likely to sell alcoholic beverages than were independently-owned pharmacies (87.5% vs. 5.4%; P < 0.001). Thirty-four (41.5%) of the 82 pharmacies that sold cigarettes and 47 (72.3%) of the 65 pharmacies that sold alcohol also displayed promotional materials for these products. NRT products were merchandised by 58% of pharmacies. Results of this study suggest that when given a choice, pharmacists choose not to sell tobacco or alcohol products. PMID- 21644022 TI - Low-income uninsured patient health and access to health care through a community health program in NC. AB - Project Access provides free primary and specialty care for low-income uninsured residents of Buncombe County, NC through a physician volunteer network. In 2010, we replicated a 1998 study investigating health issues among Project Access (PA) patients. Over 300 patients enrolled in PA in 2009 were surveyed by telephone. Currently, as compared to 1998, fewer patients were employed (31% vs. 44.2%, p < 0.05), more patients were unable to return to work (15.6% vs. 8%, p < 0.05), and patients were enrolled in PA for longer (24.5 months vs. 14 months). The SF-12 Physical Health score was worse in 2010 (p < 0.05). With the implementation of Health Reform, certain groups of people will still require PA services, but a significant portion will be covered by Medicaid; with our PA patients reporting poorer health status, Medicaid resources may be strained even more than anticipated. PMID- 21644023 TI - Legislators' beliefs on tobacco control policies in Nevada. AB - The purpose of this study was to identify Nevada legislators' views on comprehensive smoke-free (SF) policy development. The Nevada Clean Indoor Air Act (NCIAA) is a weak law that prohibits smoking in most indoor public places, excluding stand-alone bars and casino gaming areas. Nevada's state senators and assembly members were contacted to participate in the study. A literature review guided modifications of an instrument previously used to measure county-level officials' policy views in Kentucky. Descriptive statistics were conducted for selected variables, while independent t tests and one-way analysis of variance were used to examine differences between various groups. 23 of 63 legislators participated. Even though the majority of officials recognized smoking as a health hazard and nicotine as addictive, there was not overwhelming support for strengthening the NCIAA, raising cigarette excise taxes or providing cessation benefits to citizens. Officials believed that the NCIAA was having a negative economic impact on smaller gaming businesses, but not on the casino industry. Democrats were more likely than Republicans to agree that raising the excise tax by $1 is important for needed state revenues. 63% of legislators believed that they would be persuaded to strengthen the NCIAA regardless of its financial impact on small businesses, if their constituents supported such a move. No other state relies on gaming revenues as much as Nevada. Given that legislators are strongly influenced by their constituents' views, policy advocates need to establish grassroots support for strengthening the current NCIAA and also tobacco control laws in general. PMID- 21644024 TI - Exploring mediators of food insecurity and obesity: a review of recent literature. AB - One in seven American households experience food insecurity at times during the year, lack of money and other resources hinder their ability to maintain consistent access to nutritious foods. Low-income, ethnic minority, and female headed households exhibit the greatest risk for food insecurity, which often results in higher prevalence of diet-related disease. The food insecurity-obesity paradox is one that researchers have explored to understand the factors that influence food insecurity and its impact on weight change. The aim of this inquiry was to explore new evidence in associations of food insecurity and obesity in youth, adult, and elderly populations. A literature search of publication databases was conducted, using various criteria to identify relevant articles. Among 65 results, 19 studies conducted since 2005 were selected for review. Overall, the review confirmed that food insecurity and obesity continue to be strongly and positively associated in women. Growing evidence of this association was found in adolescents; but among children, results remain mixed. Few studies supported a linear relationship between food insecurity and weight outcomes, as suggested by an earlier review. New mediators were revealed (gender, marital status, stressors, and food stamp participation) that alter the association; in fact, newer studies suggest that food stamp participation may exacerbate obesity outcomes. Continued examination through longitudinal studies, development of tools to distinguish acute and chronic food insecurity, and greater inclusion of food security measurement tools in regional and local studies are warranted. PMID- 21644025 TI - Reported benefits of participation in a research study. AB - Racial and ethnic minorities are significantly underrepresented in clinical research trials. Several socio-cultural and systemic barriers, ranging from discrimination by the health care system, medical mistrust, to low physician referral rates and lack of knowledge of research studies have been identified as impacting participation. One hundred and fifteen participants were culturally matched and were interviewed followed by up to an additional four interviews over a 12 month period. Responses were analyzed to understand the perceived benefits to participating in a prospective, randomized, longitudinal clinical research trial about screening colonoscopy. Over two-thirds (64.4%) of participants reported "knowledge, awareness, and/or information about colonoscopy and general health" as being the greatest benefit they received. Desire to undergo the screening and the pride of completing the study was ranked second and third, respectively. Understanding the reasons that participants choose to participate in research studies will ultimately assist researchers close the gap in minority representation, allowing for greater generalizability of research findings. PMID- 21644026 TI - EEG microstates during resting represent personality differences. AB - We investigated the spontaneous brain electric activity of 13 skeptics and 16 believers in paranormal phenomena; they were university students assessed with a self-report scale about paranormal beliefs. 33-channel EEG recordings during no task resting were processed as sequences of momentary potential distribution maps. Based on the maps at peak times of Global Field Power, the sequences were parsed into segments of quasi-stable potential distribution, the 'microstates'. The microstates were clustered into four classes of map topographies (A-D). Analysis of the microstate parameters time coverage, occurrence frequency and duration as well as the temporal sequence (syntax) of the microstate classes revealed significant differences: Believers had a higher coverage and occurrence of class B, tended to decreased coverage and occurrence of class C, and showed a predominant sequence of microstate concatenations from A to C to B to A that was reversed in skeptics (A to B to C to A). Microstates of different topographies, putative "atoms of thought", are hypothesized to represent different types of information processing.The study demonstrates that personality differences can be detected in resting EEG microstate parameters and microstate syntax. Microstate analysis yielded no conclusive evidence for the hypothesized relation between paranormal belief and schizophrenia. PMID- 21644027 TI - Noninvasive epileptic seizure localization from stochastic behavior of short duration interictal high density scalp EEG data. AB - The stochastic behavior of the phase synchronization index (SI) in different EEG bands was examined for noninvasive localization of the epileptogenic areas from the short duration (30-60 s), seizure-free and spike-free high density (256 channel) scalp EEG data. We also examined the cross-frequency and cross-electrode coupling in different EEG bands. EEG data of four subjects was used. The seizure areas were localized with subdural recordings with an 8*8 grid electrode array. It was found that the stochastic behavior of the SI in low gamma band (30-50 Hz) was higher in epileptogenic areas. The beta (12-30 Hz) band also showed similar tendencies. The stochastic behavior in theta (3-7 Hz) band was depressed in the seizure area while it was widespread in large areas over the scalp in the alpha (7-12 Hz) band. The stochastic behavior of the cross-frequency and cross electrode couplings in theta-gamma, alpha-gamma and beta-gamma bands were decreased in the seizure areas for all four subjects. These findings suggest that it is possible to localize the epileptogenic areas from the short duration seizure-free and spike-free high density scalp EEG data. PMID- 21644028 TI - The science of training in evidence-based treatments in the context of implementation programs: current status and prospects for the future. PMID- 21644029 TI - Recent progress of Shiga toxin neutralizer for treatment of infections by Shiga toxin-producing Escherichia coli. AB - Infection with Shiga toxin (Stx)-producing Escherichia coli (STEC), including O157:H7, causes bloody diarrhea and hemorrhagic colitis in humans, occasionally resulting in fatal systemic complications, such as neurological damage and hemolytic-uremic syndrome. Because Stx is a major virulence factor of the infectious disease, a series of Shiga toxin neutralizers with various structural characteristics has been developed as promising therapeutic agents. Most of these agents function to bind to the toxin directly and inhibit the binding to its receptor present on the target cells. Other neutralizers do not inhibit receptor binding but induce aberrant intracellular transport of the toxin, resulting in effective detoxification. Such a novel type of Stx neutralizer provides a new therapeutic strategy against STEC infections. Here, recent progress of the development of Stx neutralizers is reviewed. PMID- 21644030 TI - Cancer vaccines. Any future? AB - The idea that vaccination can be used to fight cancer is not new. Approximately 100 years ago, researchers attempted to stimulate a tumor-specific, therapeutic immune response to tumors by injecting patients with cells and extracts from their own tumors, or tumors of the same type from different individuals. During the last decade, great efforts have been made to develop immunotherapeutic approaches for the treatment of malignant diseases as alternatives to traditional chemo- and radiotherapy. A quintessential goal of immunotherapy in cancer is treatment with vaccines that elicit potent anti-tumor immune responses without side effects. In this article, we have attempted to review some of the most problematic issues facing the development of cancer vaccines. With the prospect of immunosuppression, an ill-designed cancer vaccine can be more harmful than a no-benefit therapy. We have noted that "immunoediting" and "immunodominance" are the premier setbacks in peptide-based vaccines and therefore it appears necessary not only to manipulate the activity of a vast number of principal components but also to finely tune their concentrations in time and space. In the face of all these quandaries, it is at least doubtful that any reliable anti-cancer vaccine strategy will emerge in the near future. PMID- 21644031 TI - Human NK cells in acute myeloid leukaemia patients: analysis of NK cell activating receptors and their ligands. AB - Natural killer (NK) cell activation is strictly regulated to ensure that healthy cells are preserved, but tumour-transformed or virus-infected cells are recognized and eliminated. To carry out this selective killing, NK cells have an ample repertoire of receptors on their surface. Signalling by inhibitory and activating receptors by interaction with their ligands will determine whether the NK cell becomes activated and kills the target cell. Here, we show reduced expression of NKp46, NKp30, DNAM-1, CD244 and CD94/NKG2C activating receptors on NK cells from acute myeloid leukaemia patients. This reduction may be induced by chronic exposure to their ligands on leukaemic blasts. The analysis of ligands for NK cell-activating receptors showed that leukaemic blasts from the majority of patients express ligands for NK cell-activating receptors. DNAM-1 ligands are frequently expressed on blasts, whereas the expression of the NKG2D ligand MICA/B is found in half of the patients and CD48, a ligand for CD244, in only one-fourth of the patients. The decreased expression of NK cell-activating receptors and/or the heterogeneous expression of ligands for major receptors on leukaemic blasts can lead to an inadequate tumour immunosurveillance by NK cells. A better knowledge of the activating receptor repertoire on NK cells and their putative ligands on blasts together with the possibility to modulate their expression will open new possibilities for the use of NK cells in immunotherapy against leukaemia. PMID- 21644032 TI - An ADP ribosylation factor-GTPase activating protein negatively regulates the production of proinflammatory mediators in response to lipopolysaccharide. AB - An ADP ribosylation factor-GTPase activating protein (ASAP1) is highly expressed in a variety of tumor cells and is involved in the cell motility, invasion, and metastasis. In order to elucidate the involvement of ASAP1 in lipopolysaccharide (LPS)-mediated inflammatory response, the effect of ASAP1 silencing on LPS induced proinflammatory mediators production was examined by using RAW 264.7 macrophage-like cells. ASAP1 was constitutively expressed in the cells and the expression was augmented by LPS stimulation. Silencing of ASAP1 with small interfering RNA enhanced the production of tumor necrosis factor-alpha, interleukin 6, interferon-beta, and nitric oxide in response to LPS. ASAP1 silencing augmented the activation of nuclear factor (NF)-kappaB and several mitogen-activated protein kinases (MAPKs). On the other hand, ASAP1 silencing did not affect the expression of IRAK4, TRAF6, and Akt as the upstream molecules of NF-kappaB signaling. A series of toll-like receptor ligands as well as LPS augmented the ASAP1 expression. Taken together, ASAP1 was suggested to negatively regulate LPS-induced proinflammatory mediators production through down-regulating LPS signaling. The feedback function of ASAP1 in LPS-mediated inflammatory response is discussed. PMID- 21644033 TI - Involvement of damage-associated molecular patterns in tumor response to photodynamic therapy: surface expression of calreticulin and high-mobility group box-1 release. AB - Damage-associated molecular patterns (DAMPs), danger signal molecules expressed after injury or infection, have become recognized as prerequisite for orchestrating effective anti-tumor host response. The expression of two prototypical DAMPs, calreticulin and high-mobility group box-1 (HMGB1) protein, was examined following Photofrin-photodynamic therapy (PDT) of Lewis lung carcinoma (LLC) cells in vitro and LLC tumors growing in syngeneic mice. Cell surface expression of calreticulin was found to be highly increased at 1 h after PDT treatment both in vitro and in vivo. Increased exposure of calreticulin was also detected on the surface of macrophages from PDT-treated LLC tumors. At the same time interval, a rise in serum HMGB1 was detected in host mice. Intracellular staining of macrophages co-incubated for 16 h with PDT-treated LLC cells revealed elevated levels of HMGB1 in these cells. The knowledge of the involvement of these DAMPs uncovers important mechanistic insights into the development of host response induced by PDT. PMID- 21644034 TI - Prognostic role of FOXP3+ regulatory T cells infiltrating human carcinomas: the paradox of colorectal cancer. AB - The accumulation of regulatory T cells (Tregs) at high density in various human carcinomas is generally associated with a poor prognosis, as expected from their capacity to inhibit antitumor immunity. Surprisingly, in patients bearing colorectal carcinoma (CRC), high regulatory T-cell infiltration is associated with a favorable prognosis, as shown by the analysis of seven clinical studies. To explain this paradox, we emphasize a putative role of the dense microbiological flora present in the large intestine with a trend toward translocation through the tumor. This microbiological hazard requires a T-cell mediated inflammatory anti-microbial response that involves Th17 cells and can thereby promote cancer growth. This Th17-cell-dependent proinflammatory and tumor enhancing response can be attenuated by Tregs, thus constituting a possible explanation for their favorable role in CRC prognosis. The link between a high density of FOXP3-positive cells in CRC immune infiltrates and favorable prognosis should lead us to consider tumor infiltrating Tregs as allies to be respected, rather than enemies to be destroyed during trials of CRC treatment. PMID- 21644035 TI - DNA fusion vaccines enter the clinic. AB - Induction of effective immune attack on cancer cells in patients requires conversion of weak tumor antigens into strong immunogens. Our strategy employs genetic technology to create DNA vaccines containing tumor antigen sequences fused to microbial genes. The fused microbial protein engages local CD4+ T cells to provide help for anti-tumor immunity, and to reverse potential regulation. In this review, we focus on induction of CD8+ T cells able to kill target tumor cells. The DNA vaccines incorporate tumor-derived peptide sequences fused to an engineered domain of tetanus toxin. In multiple models, this design induces strong CD8+ T-cell responses, able to suppress tumor growth. For clinical relevance, we have used "humanized" mice expressing HLA-A2, successfully inducing cytolytic T-cell responses against a range of candidate human peptides. To overcome physical restriction in translating to patients, we have used electroporation. Clinical trials of patients with cancer are showing induction of responses, with preliminary indications of suppression of tumor growth and evidence for clinically manageable concomitant autoimmunity. PMID- 21644036 TI - Elevated myeloid-derived suppressor cells in pancreatic, esophageal and gastric cancer are an independent prognostic factor and are associated with significant elevation of the Th2 cytokine interleukin-13. AB - We undertook a comprehensive analysis of circulating myeloid-derived suppressor cells (MDSCs) and T regulatory cells (Tregs) in pancreatic, esophageal and gastric cancer patients and investigated whether MDSCs are an independent prognostic factor for survival. We evaluated a series of plasma cytokines and in particular re-evaluated the Th2 cytokine interleukin-13 (IL-13). Peripheral blood was collected from 131 cancer patients (46 pancreatic, 60 esophageal and 25 gastric) and 54 healthy controls. PBMC were harvested with subsequent flow cytometric analysis of MDSC (HLADR(-) Lin1(low/-) CD33(+) CD11b(+)) and Treg (CD4(+) CD25(+) CD127(low/-) FoxP3(+)) percentages. Plasma IL-2, IL-4, IL-5, IL 6, IL-10, IL-12 (p70), IL-13, IL-17, G-CSF, IFN-gamma, TNF-alpha and VEGF levels were analyzed by the Bio-Plex cytokine assay. Plasma arginase I levels were analyzed by ELISA. MDSCs and Tregs were statistically significantly elevated in pancreatic, esophageal and gastric cancer compared with controls, and MDSC numbers correlated with Treg levels. Increasing MDSC percentage was associated with increased risk of death, and in a multivariate analysis, MDSC level was an independent prognostic factor for survival. A unit increase in MDSC percentage was associated with a 22% increased risk of death (hazard ratio 1.22, 95% confidence interval 1.06-1.41). Arginase I levels were also statistically significantly elevated in upper gastrointestinal cancer patients compared with controls. There was Th2 skewing for cytokine production in all three diseases, and importantly there were significant elevations of the pivotal Th2 cytokine interleukin-13, an increase that correlated with MDSC levels. PMID- 21644037 TI - A harmonized classification system for FTLD-TDP pathology. PMID- 21644038 TI - Clinicopathologic study on an ALS family with a heterozygous E478G optineurin mutation. AB - We investigated a family manifesting amyotrophic lateral sclerosis (ALS) with a heterozygous E478G mutation in the optineurin (OPTN) gene. Clinically, slow deterioration of motor function, mood and personality changes, temporal lobe atrophy on neuroimaging, and bizarre finger deformity were noted. Neuropathologically, TAR DNA-binding protein 43 (TDP-43)-positive neuronal intracytoplasmic inclusions were observed in the spinal and medullary motor neurons. In these cells, the immunoreactivity of nuclear TDP-43 was reduced. Consecutive sections revealed that the inclusions were also reactive with anti ubiquitin and anti-p62 antibodies, but noticeably negative for OPTN. In addition, TDP-43/p62-positive glial cytoplasmic inclusions (GCIs) were scattered throughout the spinal cord and the medullary motor nuclei. Furthermore, Golgi fragmentation was identified in 70% of the anterior horn cells (AHCs). The presence of AHCs with preserved nuclear TDP-43 and a fragmented Golgi apparatus, which are unrecognizable in sporadic ALS, indicates that patients with the E4787G OPTN mutation would manifest Golgi fragmentation before loss of nuclear TDP-43. In the neocortex, GCIs were sparsely scattered among the primary motor and temporal cortices, but no neuronal TDP-43-positive inclusions were detected. In the amygdala and the ambient gyrus, argyrophilic grains and ballooned neurons were seen. The thorough neuropathologic investigations performed in this work demonstrated that OPTN-positive inclusion bodies, if any, were not prominent. We postulate that optineurinopathy is closely linked with TDP-proteinopathy and speculate that this heterozygous E478G mutation would cause ALS by acting through a dominant-negative mechanism. PMID- 21644039 TI - The relationship between plasma homocysteine level and different treatment modalities in patients with ankylosing spondylitis. AB - To determine plasma homocysteine levels in ankylosing spondylitis (AS) and their correlation with disease activity measurements. To examine the effects of different treatment modalities on homocysteine levels. One hundred eight patients diagnosed with AS according to New York criteria and 65 healthy individuals matched in terms of age and gender were enrolled in the study. Patients were given detailed physical examinations. The Bath AS Disease Activity Index (BASDAI) was used for disease activity, the Bath AS Metrology Index (BASMI) for spinal mobility, the Bath AS Functional Index (BASFI) to determine functional status and the Bath AS Radiological Index (BASRI) for radiological analysis. Sedimentation rate (ESR) and C reactive protein (CRP) levels, acute phase reactants, were measured. Plasma homocysteine levels, serum interleukin (IL) -6 and serum tumor necrosis factor- alpha (TNF- alpha) levels were investigated using the enzyme linked immunosorbent assay (ELISA) method. Plasma homocysteine levels in AS patients were statistically significantly lower compared with those in the healthy control group (P = 0.0001). There was no significant difference among sub groups established on the basis of medical treatments and disease activity (BASDAI <=4 and >4). No statistically significant correlation was determined between homocysteine level and disease activity parameters, radiological score and functional indices. A significant negative correlation was, however, established between age and homocysteine level in the AS group (P < 0.05, r = 0.426). Plasma homocysteine was lower in AS patients compared with the control group. This is not correlated with disease activation and medical treatment employed. PMID- 21644040 TI - Frequency of musculoskeletal complications among the patients receiving solid organ transplantation in a tertiary health-care center. AB - Solid organ transplantation is an important lifesaving procedure mainly performed in patients with end-stage organ failure such as liver cirrhosis, congestive heart failure, and end-stage renal disease. While these complications are among the most preventable adverse effects of solid organ transplantation, these are generally neglected by physicians. Accordingly, this study was performed to evaluate the frequency of musculoskeletal complications among the patients receiving solid organ transplantation in a tertiary health-care center in a developing country. This cohort study was performed from 2000 to 2009, among fifty patients receiving organ transplantation (liver, heart, and lung) attending to a training hospital in Tehran, Iran. The main variables were musculoskeletal complaints and findings that were measured according to patients' self-report and clinical examination. The mean age of the patients was 40.2 +/- 10.9 years ranging from 5 to 58 years. Twenty out of 50 patients (40%) had musculoskeletal complaints that the most common complaint was the arthralgia. Also, the mechanical arthritis was the most common clinical finding in clinical examination (24%). Low serum level of vitamin D (74.4%) and high serum alkaline phosphatase level (27.9%) were the most common biochemical abnormalities in understudy population. Finally, it may be concluded that nearly forty percent of patients receiving solid organ transplantation may develop musculoskeletal findings and/or complaints. These complications may be found and treated with regular examinations to reduce the burden of disease. PMID- 21644041 TI - Cytokine modulators in the treatment of sarcoidosis. AB - Sarcoidosis is a granulomatous lung disease in which several cytokines play a pivotal pathogenetic role. Steroid-resistant disease can be treated with immunosuppressive drugs, antimalarial therapies and recently with anti-TNFalpha agents. The use of biological agents for the treatment of sarcoidosis springs from research into the pathogenesis of the disease and also from the experience of rheumatologists with other chronic inflammatory diseases. Rituximab, golimumab and ustekinumab are cytokine modulators, useful in the treatment of immunoinflammatory disorders, for which randomized trials to evaluate safety and efficacy in sarcoidosis are not yet available. Novel anticytokine drugs administered alone or in association may offer a new approach to treatment of the disease. This review focuses on recent advances in anti-TNFalpha agents and cytokine modulators for the treatment of sarcoidosis and their therapeutic prospects. PMID- 21644042 TI - Using TNF-alpha antagonist adalimumab for treatment for multisystem sarcoidosis: a case study. AB - We report the usage of the TNF-alpha antagonist adalimumab in patients with progressive multisystem sarcoidosis. Three patients with multisystem sarcoidosis (MSS) were treated with adalimumab for 12 months. All three patients were quickly responded to adalimumab and experienced a nearly complete regression of the symptoms that lead to an intensive immunosuppression. However, some accompanying symptoms of sarcoidosis, such as splenomegalia, did not respond. One patient suffered 18 months later a new unspecified abdominal lymphadenopathy. TNF-alpha antagonists can be helpful agents in the treatment for MSS. However, the experience with TNF-alpha antagonists in patients with sarcoidosis is still limited. Multicenter trials and a comparison of the different agents are needed to validate the safety and efficacy in these patients. Optimal dosage, duration of therapy and long-term toxicity of anti-TNF therapy in patients with refractory sarcoidosis are yet to be determined in prospective trials. PMID- 21644043 TI - An unusual pediatric case with neurofibromatosis and systemic lupus erythematosus. AB - Neurofibromatosis type 1 (NF1) is a relatively common autosomal dominant disorder affecting mainly ectodermal and mesodermal tissues. It is well known that patients with NF1 have an increased risk of developing benign and malignant tumors, but its association with autoimmune diseases has been rarely reported. Systemic lupus erythematosus is an autoimmune chronic inflammatory disease that has the potential to affect various organ systems. There are four cases with NF1 and SLE reported in the literature up to date. Here, we report a 9-year-old girl presenting with NF1 and SLE, and to our knowledge, this is the first childhood case in the literature. PMID- 21644044 TI - The role of human xanthine oxidoreductase (HXOR), anti-HXOR antibodies, and microorganisms in synovial fluid of patients with joint inflammation. AB - This work is to investigate the levels of human xanthine oxidoreductase (HXOR), its antibodies, and microorganisms in synovial fluid of patients with untreated rheumatoid joint diseases. Synovial fluids were collected from sixty-four patients with rheumatoid joint diseases. Sixty-four age-matched individuals were included as control. Xanthine oxidoreductase (XOR) proteins level and anti-XOR antibodies were determined in the blood and synovial fluid, using human XOR as antigen, by enzyme-linked immunosorbent (ELISA) assay. Synovial fluids were cultured for bacteria and fungi. The titers of XOR protein in the synovial fluid of patients with rheumatoid arthritis were 90.43 +/- 23.37 MUg/ml (mean +/- SD, n = 29) and up to 62.42 +/- 8.74 MUg/ml (mean +/- SD, n = 35) in other joint inflammation. Anti-HXOR antibodies titers in patients were 167.72 +/- 23.64 MUg/ml, n = 64, which was significantly higher in rheumatoid arthritis patients. The results indicated that anti-HXOR antibodies in synovial fluids have a protective role as high concentrations against XOR were detected in inflammatory arthritis. These antibodies play a role in eliminating XOR from synovial fluids. However, immune complex formation could activate complement and participate in propagating the inflammatory cycle. Synovial aspirate ordinary microbial cultures were negative for any bacteria or fungi, but that does not exclude organisms of special culture requirements. PMID- 21644045 TI - A comparative analysis of serological parameters and oxidative stress in osteoarthritis and rheumatoid arthritis. AB - Progression of rheumatoid arthritis (RA) and osteoarthritis (OA) is associated with inflammation and oxidative stress. Previous studies have shown that there was no difference between RA and OA patients regarding the percentages of the different lymphocytes subsets reflecting the abnormalities in T cells and its subsets that may contribute to the pathogenesis of OA as in RA. Therefore, the present study was aimed to analyze that whether disease activity of OA is able to affect a few serological and biochemical parameters in the same way as RA does or differently. The study was done on 36 asymptomatic controls (25 women), 28 patients with OA (20 women), 36 patients with RA (22 women). Patients with OA were screened according to radiological and clinical finding of Kellgren and Lawrence grade and ACR criteria and assessed by VAS and WOMAC score. Patients with RA were selected who were fulfilling 4/5 symptoms of ACR criteria, and their DAS28-CRP, VAS score, and RF positivity were evaluated. Participants of the groups were matched for sex, age, weight, and height (body mass index). The BMI of all three groups was also found to be the same (P > 0.05). The mean level of LDL, cholesterol, MDA, CRP, and triglyceride was significantly (P < 0.05 or P < 0.01) higher in both OA and RA as compared to control. The mean level of total lipid, cholesterol, MDA, CRP, and triglyceride was found to be significantly (P < 0.05 or P < 0.01) higher in RA as compared to OA. The pre-treatment CRP level of both groups of patients showed significant and direct relation with total lipid (r = 0.27, P < 0.05) and cholesterol (r = 0.66, P < 0.01). Inverse relation was observed between uric acid and creatinine (r = -0.26, P < 0.05) and cholesterol and HDL (r = -0.34, P < 0.01). Our study shows the similar trend in lipid profile and other parameters studied in both patients with OA and patients with RA with more pronounced changes in RA. PMID- 21644046 TI - Evaluation of liver function tests in scleroderma patients. AB - Systemic sclerosis is a clinically heterogeneous, systemic disorder which affects the connective tissue of the skin, internal organs, and the walls of blood vessels. It is characterized by alterations of the microvasculature, disturbances of the immune system and by massive deposition of collagen and other matrix substances in the connective tissue. This study was done to evaluate the frequency of liver disease in patients with scleroderma and, secondarily, to study the frequency of infection of hepatitis B and C virus in these patients and determine frequency of serum auto-antibodies in this disease. We studied patients with scleroderma, localized or systemic, in the outpatient clinic of rheumatology and dermatology departments, at King Khalid University Hospital. As for a comparison, healthy persons coming to the clinic with the same mean age were considered as control group. Forty patients with the diagnosis of scleroderma included in this work, 35% had elevated gamma-glutamyl-transferase (gamma-GT), 30% had elevated alkaline phosphatase (AP) and in 17.5%, the alanine-amino transferase (ALT) was above the reference values. The ALT had changed to be more in scleroderma patients than in controls. Twenty percent (20%) of the patients tested positive for anti-smooth muscle antibodies (anti-SMA) and only one patient had anti-mitochondrial antibodies (AMA). There was no statistical difference between the two groups regarding antibody testing. Anti-HCV antibodies were observed in one patient, and HBsAg was detected in another scleroderma patient. There was no patient with clinically significant hepatic disease. In this study, although changes in liver enzymes in patients with scleroderma were not uncommon, there was no scleroderma patient with clinical manifestations of liver disease. PMID- 21644047 TI - Bone morphogenetic protein receptor IB as a marker for enrichment of osteogenic precursor-like cells in human dermis. AB - The scarcity of bone marrow mesenchymal stromal cells (BMSCs) prompts the search for alternative sources for cell-based bone defects repair. Human dermal fibroblasts (FBs) have been shown to have a high proliferative potential and the capacity to differentiate into an osteogenic phenotype. The easy and repeated harvest in large quantities makes this cell source a potential candidate for bone tissue engineering. The aim of our study was to compare directly the immune phenotype, proliferative capacity and osteogenic differentiation potential of FBs with that of "gold standard" BMSCs or adipose-derived mesenchymal stromal cells (ADSCs), another alternative osteoprogenitor cell source. Flow cytometry demonstrated that FBs, ADSCs and BMSCs shared common cell surface marker protein expression profiles when using a panel of surface antigens. FBs had the highest proliferative potential, but lowest osteogenic differentiation potential in vitro, compared with ADSCs or BMSCs. More importantly, BMPR-IB(+)-sorted FBs subpopulation had a higher osteogenic differentiation potential than BMPR-IB(-) sorted FBs subpopulation. Our results indicated that the heterogeneous FBs were not an appropriate cell source for bone tissue engineering. Immunoselection by BMPR-IB can generate highly purified osteogenic precursor-like cells in the human dermis. PMID- 21644048 TI - Levels of TGF-beta(1) in serum and culture supernatants of CD4(+)CD25 (+) T cells from patients with non-segmental vitiligo. AB - Compelling evidences support an autoimmune basis of non-segmental vitiligo, and dysregulation of CD4(+)CD25(+) regulatory T cell (Treg) is assumed to contribute to the pathogenesis of vitiligo. Serum levels of transforming growth factor-beta (TGF-beta), an important immunoregulatory cytokine produced by Treg cells, has been reported significantly decreased in patients with vitiligo. However, relation between the decrease in TGF-beta and the dysfunction of Treg cells in pathogenesis of vitiligo was still undemonstrated. To further reveal the role of TGF-beta in vitiligo, 46 patients with non-segmental vitiligo and 25 age- and sex matched healthy control subjects were enrolled in the study. CD4(+)CD25(+) T cells isolated from peripheral venous blood with a CD4(+)CD25(+) regulatory T cell isolation kit were cultured with or without anti-CD3 mAbs and anti-CD28 mAbs for 4 days. The TGF-beta1 levels in serum and culture supernatants were detected by enzyme-linked immunosorbent assay in both groups. We have found that the TGF beta1 levels both in serum and culture supernatants in the presence of anti-CD3 mAbs and anti-CD28 mAbs were decreased in the active vitiligo group when compared with the control group or stable vitiligo group, and were negatively correlated with the percentage of involved body area. These results suggested that TGF-beta may play a role in the pathogenesis of non-segmental vitiligo related to the suppressive function of Tregs. PMID- 21644049 TI - Evaluation of basal cell carcinoma microcirculation through orthogonal polarization technique. AB - Basal cell carcinomas are the most common cutaneous neoplasias. They rarely metastasize, but can cause clinically significant local destruction and disfigurement if neglected or inadequately treated. The spectral imaging technique, a recently developed method of orthogonal light polarization, allows in vivo transcutaneous evaluation of tumor microcirculation. The objective of this work was to compare tumor and normal skin microcirculation in patients using the orthogonal polarization (OPS) technique. Forty histologically confirmed basal cell carcinomas of 36 patients were evaluated in order to accomplish our objective. Compared to control skin, basal cell carcinomas had significantly greater capillary diameters (10.6 +/- 2.6 and 18.5 +/- 6.2 MUm, respectively), capillary density (25.0 +/- 10.4 and 50.3 +/- 16.9 cm/cm(2), respectively) and red blood cell velocities (0.06 +/- 0.03 and 0.11 +/- 0.07 mm/s). Our results have demonstrated significant microcirculatory alterations in skin tumors using the OPS technique that allows its transcutaneous evaluation in vivo, in the absence of anesthesia and invasive surgical procedures. This technique could be a promising tool for researchers studying diseases in which angiogenesis plays a major role. PMID- 21644050 TI - Evaluating rational non-cross-resistant combination therapy in advanced clear cell renal cell carcinoma: combined mTOR and AKT inhibitor therapy. AB - PURPOSE: Inhibition of the mammalian target of rapamycin (mTOR), a regulator of hypoxia inducible factor (HIF), is an established therapy for advanced renal cell cancer (RCC). Inhibition of mTOR results in compensatory AKT activation, a likely resistance mechanism. We evaluated whether addition of the Akt inhibitor perifosine to the mTOR inhibitor rapamycin would synergistically inhibit RCC. METHODS: Select RCC cell lines were studied [786-O, A498 (VHL mutant), CAKI-1 (VHL wild type), and 769-P (VHL methylated)] with single agent and combination therapy. Growth inhibition was assessed by MTT and cell cycling by flow cytometry. Phospho-AKT (S473) and HIF-2alpha were assessed by Western blot. Total RNA was isolated from 786-O cells subjected to single agent and combination treatments. In these cells, genome-wide expression profiles were assessed, and real-time PCR was used to confirm a limited set of expression results. RESULTS: Three out of four cell lines (CAKI-1, 769-P, and 786-O) were sensitive to single agent perifosine with 50% inhibitory concentrations ranging from 5 to 10 MUM. Perifosine blocked phosphorylation of AKT induced by rapamycin and inhibited HIF 2alpha expression in 786-O and CAKI-1. Combined treatment resulted in sub additive growth inhibition. GeneChip analysis and pathway modeling revealed inhibition of the IL-8 pathway by these agents, concomitant with up-regulation of the KLF2 gene, a known suppressor of HIF1alpha. CONCLUSIONS: Perifosine is active in select RCC lines, abrogating the induction of AKT phosphorylation mediated by mTOR inhibition. Combined mTOR and AKT inhibition resulted in the modulation of pro-angiogenesis pathways, providing a basis for future investigations. PMID- 21644051 TI - Epidemiology and predictors of spinal injury in adult major trauma patients: European cohort study. AB - This is a European cohort study on predictors of spinal injury in adult (>=16 years) major trauma patients, using prospectively collected data of the Trauma Audit and Research Network from 1988 to 2009. Predictors for spinal fractures/dislocations or spinal cord injury were determined using univariate and multivariate logistic regression analysis. 250,584 patients were analysed. 24,000 patients (9.6%) sustained spinal fractures/dislocations alone and 4,489 (1.8%) sustained spinal cord injury with or without fractures/dislocations. Spinal injury patients had a median age of 44.5 years (IQR = 28.8-64.0) and Injury Severity Score of 9 (IQR = 4-17). 64.9% were male. 45% of patients suffered associated injuries to other body regions. Age <45 years (>=45 years OR 0.83 0.94), Glasgow Coma Score (GCS) 3-8 (OR 1.10, 95% CI 1.02-1.19), falls >2 m (OR 4.17, 95% CI 3.98-4.37), sports injuries (OR 2.79, 95% CI 2.41-3.23) and road traffic collisions (RTCs) (OR 1.91, 95% CI 1.83-2.00) were predictors for spinal fractures/dislocations. Age <45 years (>=45 years OR 0.78-0.90), male gender (female OR 0.78, 95% CI 0.72-0.85), GCS <15 (OR 1.36-1.93), associated chest injury (OR 1.10, 95% CI 1.01-1.20), sports injuries (OR 3.98, 95% CI 3.04-5.21), falls >2 m (OR 3.60, 95% CI 3.21-4.04), RTCs (OR 2.20, 95% CI 1.96-2.46) and shooting (OR 1.91, 95% CI 1.21-3.00) were predictors for spinal cord injury. Multilevel injury was found in 10.4% of fractures/dislocations and in 1.3% of cord injury patients. As spinal trauma occurred in >10% of major trauma patients, aggressive evaluation of the spine is warranted, especially, in males, patients <45 years, with a GCS <15, concomitant chest injury and/or dangerous injury mechanisms (falls >2 m, sports injuries, RTCs and shooting). Diagnostic imaging of the whole spine and a diligent search for associated injuries are substantial. PMID- 21644052 TI - Introduction to the interaction between gonadal steroids and the central nervous system. AB - The sex steroids are frequently referred to as the gonadal steroids and are erroneously assumed to be exclusively linked to the ovaries in women or the testes in men and the functions of the reproductive tract. This chapter will provide an overview of some of the extragonadal effects of these hormones, focusing on the central nervous system, and the mechanisms of hormone action. Hormone synthesis and metabolism within the CNS will be discussed with particular focus on the role of aromatase. Sex steroids exert many of their effects via intracellular receptors and these genomic responses tend to be slow in onset, however, some responses to steroids occur more quickly and are mediated via membrane receptors and involve interactions with many different transduction pathways to produce a diverse array of responses. These complexities do pose challenges but also offer opportunity for novel approaches for therapeutic exploitation as the pharmacological tools with which to modulate systems become increasingly available. PMID- 21644053 TI - Transapical septal occluder for paravalvular leakage of the mitral valve: a novel method for interventional treatment in high-risk patients. PMID- 21644054 TI - PARP1 and CASP3 gene expression in a patient with multiple head and neck squamous cell carcinoma and Parkinson disease. PMID- 21644055 TI - Recent invasion and low level of divergence between diploid and triploid forms of Carassius auratus complex in Croatia. AB - Carassius auratus is an invasive species in European waters, comprising a complex of diploid and polyploid forms with different modes of reproduction. However, the evolutionary history and relationships between the diploids and polyploids are still unresolved. In this study, 51.5% diploids and 48.5% triploids, including four triploid males, were discovered among the 363 individuals sampled in Croatia. We used eight microsatellite loci and mitochondrial displacement loop sequences to analyze the structure and origin of populations; and to attempt to infer the evolutionary history of the two different forms in Croatia. Microsatellite analyses revealed high allelic and clonal diversity, corroborating that high propagule vectors can compensate for the negative effects of genetic bottlenecks in successful invasive species. The absence of significant population structuring confirmed recent origin and rapid spreading of populations. No evidence was found for the existence of native European populations. Distances between individuals using both nuclear and mtDNA markers revealed the absence of substantial clustering on the ploidy level, while the split between the different ploidies on population level was only partial, suggesting that the reproductive isolation between the two forms is either of a very recent origin, or that there exists uni-, or bidirectional gene flow between the diploid and triploid forms. PMID- 21644056 TI - 1H, 13C, and 15N NMR assignments for the helicase interaction domain of Staphylococcus aureus DnaG primase. AB - The interaction between DnaG primase and DnaB helicase is essential for stimulating primer synthesis during bacterial DNA replication. The interaction occurs between the N-terminal domain of helicase and the C-terminal domain of primase. Here we present the (1)H, (13)C, and (15)N backbone and side-chain resonance assignments for the C-terminal helicase interaction domain of Staphylococcus aureus primase. PMID- 21644057 TI - The differential relationship between fat mass and bone mineral density by gender and menopausal status. AB - Osteoporosis and obesity are important public health problems in an aging society. We investigated the differential impacts of fat on bone mineral density (BMD) according to gender and menopausal status. We analyzed the baseline data of an ongoing observational cohort study, including a total of 502 healthy subjects 20-88 years of age (144 men, 159 premenopausal women, 199 postmenopausal women). Body composition and fat mass were measured using computed tomography and dual energy X-ray absorptiometry (DXA). BMD was measured at lumbar spines using DXA. In men and postmenopausal women, there was no significant correlation between fat and bone parameters after adjusting for age and body weight. However, in premenopausal women, BMD had significant negative correlations with waist circumference, total fat area, subcutaneous fat area, appendicular fat mass and percentage fat mass after adjusting for age and body weight. Furthermore, only in premenopausal women, the subjects with the highest quartile of percentage fat mass had the lowest BMD even after adjusting for confounding factors including age, body weight, physical activity, alcohol use and smoking history. Multiple linear regression analysis showed that percentage fat mass was a significant negative decisive factor for BMD in premenopausal women. Our study showed the differential relationship between fat mass and BMD according to gender and menopausal status. Only in premenopausal women did fat mass have a significant negative effect on bone mass. This result suggests the importance of reducing fat mass in order to achieve peak bone mass in young adult women. PMID- 21644058 TI - The incidence and residual lifetime risk of osteoporosis-related fractures in Korea. AB - Although the Korean population does not have high risk for osteoporosis, the numbers of osteoporosis-related fractures represent a considerable economic burden to society. The purpose of this study was to determine the incidence and residual lifetime risk of osteoporosis-related fractures in Korea, using data from the Health Insurance Review and Assessment Service (HIRA), which includes nationwide information compiled by the Korean government. All new visits or admissions to Korean clinics or hospitals for fractures were recorded prospectively in a nationwide cohort by the Korean HIRA using the International Classification of Diseases, 10th revision, codes and procedure codes. These data were retrospectively evaluated to determine the incidence and residual lifetime risk of osteoporosis-related fractures (hip, spine, distal radius, and humerus fractures), in men and women aged 50 years or more between 2005 and 2008. The annual incidences of osteoporosis-related fractures were 1,661, 1,646, 1,623, and 1,614 per 100,000 person-years in men and women aged 50 years or more from the year 2005 to 2008. The annual incidence of osteoporosis-related fracture in women was three times that of men. The incidence of osteoporosis-related fractures increased with advancing age. In Korea, at the age of 50 years, the residual lifetime probabilities of osteoporosis-related fractures are 59.5% for women and 23.8% for men. This study presents the baseline data for treatment and research on osteoporosis and provides an estimate of osteoporosis-related fractures in Korea. PMID- 21644060 TI - Stent selection for both biliary and pancreatic strictures caused by chronic pancreatitis: multiple plastic stents or metallic stents? AB - Endoscopic stenting is an effective treatment option in the management of both benign biliary strictures and pancreatic ductal strictures. Plastic stents and self-expandable metal stents have been used with variable success for the management of both benign biliary strictures and pancreatic ductal strictures caused by chronic pancreatitis. Fully covered self-expandable metal stents of improved design represent a major technological advance which has added to the endoscopic armamentarium. Both multiple plastic stents and covered self expandable metal stents have shown promising results. However, data to support the use of self-expandable metal stents over multiple plastic stents or vice versa are still lacking. PMID- 21644059 TI - Additive effects of a cholinesterase inhibitor and a histamine inverse agonist on scopolamine deficits in humans. AB - RATIONALE: Enhancement of histaminergic neurotransmission or histaminergic plus cholinergic neurotransmission may represent novel strategies for improving cognition in Alzheimer's disease. OBJECTIVE: To evaluate the effects of a novel histamine H3 receptor inverse agonist (MK-3134), an acetylcholinesterase inhibitor (donepezil), and their combination in attenuating the cognitive impairment associated with scopolamine. METHODS: Thirty-one subjects were randomized, and 28 completed this double-blind, placebo-controlled, five-period crossover study. Cognition was assessed using the Groton Maze Learning Task (GMLT) as the primary outcome measure. The two primary hypotheses were that donepezil 10 mg and MK-3134 25 mg, respectively, would attenuate scopolamine (0.5 mg)-induced impairment as measured by the GMLT over the first 12 h after scopolamine administration (AUC(1-12) (h)). A secondary hypothesis was that the combination of donepezil and MK-3134 would attenuate scopolamine-induced cognitive impairment to a greater extent than either agent alone as measured by the GMLT AUC(1-12 h). RESULTS: The primary and secondary hypotheses were not met. Upon examining the time course of the scopolamine effects (an exploratory objective), peak effects were generally observed around 2 h after scopolamine administration. Administration of MK-3134 or donepezil improved performance on the GMLT at the 2-h time point, rather than AUC(1-12 h), compared with scopolamine alone. Moreover, it appeared that the combination of MK-3134 and donepezil blunted the scopolamine effect to a greater extent than either drug alone. CONCLUSIONS: Exploratory analyses provide evidence for cognitive improvement through inverse agonism of the H3 histamine receptor and for cooperation between human cholinergic and histaminergic neurotransmitter systems. (ClinicalTrials.gov trial registration number: NCT01181310). PMID- 21644061 TI - Evaluation of pancreatic intraepithelial neoplasia and mucin expression in normal pancreata. AB - BACKGROUND/PURPOSE: It has been suggested that pancreatic ductal adenocarcinoma (PDAC) and pancreatic intraepithelial neoplasia (PanIN) are closely related, but several reports indicate PanIN lesions can also be found in normal pancreata (normal PanINs). We examined differences in mucin expression between normal PanIN lesions and PanINs in PDACs (PDAC PanINs). METHODS: We examined 54 autopsied normal pancreata and eight autopsied PDACs for PanIN lesions; graded the pancreata specimens as PanIN-1A (non-papillary hyperplasia), PanIN-1B (papillary hyperplasia), PanIN-2 (atypical hyperplasia) or PanIN-3 (carcinoma in situ); and tested the PanIN lesions for expression of MUC1 (pan-epithelial membrane associated mucin) and MUC5AC (gastric secretory mucin) which were both previously detected in PDACs. RESULTS: In normal PanIN-1A, PanIN-1B and PanIN-2 specimens, MUC1 was expressed in 2.8, 10.5 and 9.1%, respectively, compared to 19.1, 27.6 and 13.0% in PDAC PanIN-1A, PanIN-1B and PanIN-2 specimens, respectively. MUC5AC was expressed in 41.0, 65.7 and 36.4% of normal PanIN-1A, PanIN-1B and PanIN-2 specimens, respectively, and in 80.9, 75.8 and 78.3% of PDAC PanIN-1A, PanIN-1B and PanIN-2 specimens, respectively. Differences in the frequency of MUC1 expression were significant between normal and PDAC PanIN-1A (p < 0.0001) and PanIN-1B (p < 0.05); and differences in the frequency of MUC5AC expression were significant between normal and PDAC PanIN-1A (p < 0.0001) and PanIN-2 (p < 0.05). CONCLUSIONS: Normal PanIN and PDAC PanIN lesions differed in the rates of MUC1 and MUC5AC expression. PMID- 21644062 TI - Outcomes of switching anti-TNF drugs in rheumatoid arthritis--a study based on observational data from the Finnish Register of Biological Treatment (ROB-FIN). AB - The aim of this study was to assess, based on observational data from the Finnish Register of Biological Treatment, the outcomes of switching an initial tumor necrosis factor (TNF) blocker to another in the treatment of rheumatoid arthritis (RA). RA patients, who started biological therapy with a TNF blocker between May 1999 and April 2009 and who switched to another TNF blocker, were studied (n=479). The outcomes were assessed according to the reason for and type of the switch. Outcome assessments included American College of Rheumatology 50 responder index (ACR50) response at 3 months after the switch, treatment duration of the second TNF blocker, and swollen joint counts, CRP and DAS28 score at the 3 months, best and last observations of the first and second TNF blocker, respectively. In those who switched due to lack of effectiveness (LOE), the disease activity parameters fell significantly from baseline upon use of infliximab or adalimumab, but had increased prior to the switch. Switching to another TNF blocker (etanercept or adalimumab) restored the response initially achieved with the first TNF blocker. The disease activity parameters fell significantly from baseline upon use of etanercept, and were maintained but not further improved after switching to adalimumab. TNF blocker switching seemed to be most beneficial in secondary LOE (defined as loss of ACR50 response). In those who switched due to adverse events (AE) or other reasons, a similar degree of response as had been achieved with the first agent was also achieved and maintained with the second agent. The results suggest that a second TNF blocker can restore the response in cases of secondary LOE and maintain it after switching due to an AE. PMID- 21644063 TI - [Comparative evaluation of in vitro activities of carbapenems against gram negative pathogens: Turkish data of COMPACT study]. AB - The aim of this study was to determine the in vitro activities of doripenem, imipenem, and meropenem against clinical gram-negative isolates. A total of 596 clinical isolates were obtained from intensive care unit (ICU) and non-ICU patients in 10 centers over Turkey between September-December 2008. The origin of the isolates was patients with nosocomial pneumonia (42.4%), bloodstream infections (%40.4), and complicated intraabdominal infections (17.1%). Of the isolates, 51.8% were obtained from ICU patients. The study isolates consisted of Pseudomonas spp. in 49.8%, Enterobacteriaceae in 40.3%, and other gram-negative agents in 9.9%. The minimum inhibitory concentrations (MIC) for doripenem, imipenem and meropenem were determined for all isolates in each center using Etest(r) strips (AB Biodisk, Solna, Sweden). Of the isolates, 188 (31.5%) were resistant to at least one of the carbapenems. MIC50 of doripenem against Pseudomonas spp. Was 1 mg/L which was similar to that of meropenem and two-fold lower than imipenem. Susceptibility to carbapenems in P.aeruginosa was 64% for doripenem at an MIC level of 2 mg/L, 53.9% and 63% for imipenem and meropenem at an MIC level of 4 mg/L, respectively. Doripenem and meropenem showed similar activity with the MIC90 of 0.12 mg/L whereas imipenem was four-fold less active at 0.5 mg/L. Against other gramnegative pathogens, mostly Acinetobacter spp., MIC50 was 8 mg/L for doripenem and 32 mg/L for other two carbapenems. P.aeruginosa isolates were inhibited 84.2% with doripenem and 72.1% with meropenem at the MIC level of 8 mg/L. Doripenem generally showed similar or slightly better activity than meropenem and better activity than imipenem against pathogens collected in this study. Against Pseudomonas spp., doripenem was the most active of the three carbapenems. Doripenem and meropenem were equally active against Enterobacteriaceae and at least four-fold more active than imipenem. It was concluded that doripenem seemed to be a promising agent in the treatment of nosocomial pneumonia, blood stream infections and intraabdominal infections particularly in patients who were under risk of developing antimicrobial resistance. PMID- 21644064 TI - [Investigation of Salmonella serotype Enteritidis isolates by plasmid profile analysis and pulsed field gel electrophoresis]. AB - In this study a total of 122 Salmonella serotype Enteritidis stock strains selected from the culture collection of Enterobacteriaceae Laboratory of Ankara University Faculty of Medicine, Department of Medical Microbiology, were investigated by plasmid profile analysis with the method defined by Kado and Liu and pulsed field gel electrophoresis (PFGE) according to World Health Organization protocols using SpeI and XbaI macrorestriction enzymes, for better understanding of the molecular epidemiology of S. Enteritidis. The study strains were selected from a collection of previously isolated epidemic (n= 13) and sporadic (n= 109) strains (103 stool, 16 blood and one each bile, urine and cerebrospinal fluid) obtained from 10 different cities after the year 2000. PFGE patterns were analyzed with Gene Directory software (Syngene, UK) and a similarity index was determined by using Dice coefficient and the unweighted pair group method with mathematical averaging (UPGMA). Plasmid-carrying 110 (90%) strains that harbored 1-4 plasmids with sizes ranging from 2.0 to 100 kb were separated into patterns more than 14 (p1-p14). A total of 85 (69.7%) isolates harbored the 57 kb plasmid solely or in combination with other plasmids. By PFGE, 11 distinct patterns were shown with each enzyme SpeI and XbaI. S. Enteritidis strains after digestion with macrorestriction enzyme SpeI generated 11 different PFGE patterns (A to K), whereas XbaI generated also 11 different PFGE patterns (a to k). PFGE pattern A consisted of 93 strains (76.2%) after digestion with macrorestriction enzyme SpeI, while PFGE pattern a consisted 53 (43.4%) and PFGE pattern b 42 strains (34.4%) after digestion with macrorestriction enzyme XbaI. Using two macrorestriction enzymes two PFGE cluster profiles Aa (50 strains, 40.9%) and Ab (42 strains, 34.4%) were found to be predominating among 17 different PFGE clusters. Our results confirmed the clonal nature of S. Enteritidis strains in Turkey. The use of two enzymes in PFGE analysis appeared to increase the discriminatory power of PFGE, leading to greater diversity among strains. PFGE analysis performed by SpeI and XbaI enzymes combined with plasmid profiling could be established as a useful tool for detection of genetic relationship between isolates. PMID- 21644065 TI - [Investigation of Helicobacter pylori iceA1 and iceA2 genes in patients with chronic gastritis and gastric cancer]. AB - Several virulence factors of Helicobacter pylori play crucial role in the pathogenesis of the infections.H.pylori iceA gene which is induced by the contact with epithelium during the attachment of bacterium to the gastric mucosa, possess two variants (iceA1 and iceA2). Although there are some data indicating the relationship between H.pylori iceA1 and peptic ulcer, this concept is still controversial. The aims of this study were to investigate the presence and prevalence of H.pylori iceA1 and iceA2 gene regions in the tissue samples of patients diagnosed as chronic gastritis and gastric cancer, and to evaluate whether any correlation existed between these genotypes and clinical manifestations. A total of 109 tissue samples obtained from chronic gastritis (n= 55) and gastric cancer (n= 54) patients whose H.pylori infections have been confirmed by histopathologic examination of biopsy samples, were included in the study. The presence of H.pylori in the samples were also confirmed by amplification of the ureA gene region by inhouse polymerase chain reaction (PCR). H.pylori iceA1 and iceA2 genes were directly genotyped with the use of specific primers in the gastric biopsy specimens by PCR. The total positivity rates of iceA1 and ice- A2 genotypes in patients were found as 58% (63/109) and 24% (26/109), respectively. With the special attention to chronic gastritis and gastric cancer patients, the frequencies of iceA1 gene were 51% (28/55) and 65% (35/54), while the frequencies of iceA2 gene were 20% (11/55) and 28% (15/54), respectively. The difference of positivity rates of iceA1 and iceA2 genotypes between the patient groups were not statistically significant (p> 0.05). There was also no statistically significant correlation between the genotypes and clinical manifestation (r> 0.01). As a result, H.pylori iceA1 genotype was predominant (58%) in chronic gastritis and gastric cancer patients in our region, however the prevalence of iceA2 genotype was lower (24%) similar to those data reported in the literature. Our results supported the concept that iceA gene reflects geographical differences rather than determining the clinical picture and virulence. In conclusion, multicenter and large scaled studies are needed for better evaluation of H.pylori iceA gene and disease relationship. PMID- 21644066 TI - [A water-borne tularemia outbreak caused by Francisella tularensis subspecies holarctica in Central Anatolia region]. AB - In this study, we investigated a waterborne tularemia outbreak occured in Kadiozu, a village of Cerkes county of Cankiri province (located in North-west part of central Anatolia, Turkey) between 18 November 2009-24 December 2009. Active surveillance was conducted to determine clinical characteristics and risk factors of cases after two patients from the same village had been diagnosed as oropharyngeal tularemia. All villagers were examined, and clinical specimens from cases and water samples which may be the source of outbreak in the field investigations were taken. Cases were in the form of oropharyngeal, glandular and pneumonic. Polymerase chain reaction (PCR) and cultures were conducted from lymph node aspirates, throat swabs taken from cases and samples from water sources of epidemic zone. All serum samples taken from the villagers were screened for F.tularensis antibodies with microagglutination test (MAT). Oropharyngeal tularemia was diagnosed in 11 patients, glandular form in 3 patients and pneumonic form in one patient according to clinical and laboratory results. Age of the patients ranged between 6-75 years old (mean age: 52.5 years) and thirty one of them (54.7%) were female. MAT titers ranged between 1/160 and 1/5120 in cases of tularemia. Causative agent was grown in the cultures of two patients (including a throat swab and a lymph node aspirate). F.tularensis DNA was shown by PCR in a throat swab and four lymph node aspirates. F.tularensis was also detected by PCR in the water sample obtained from one of the spring water commonly used by villagers. Only one of the lymph node samples obtained from two different patients, was positive by direct fluorescent antibody method. Causative agent was defined as F.tularensis subsp. holarctica by conventional and also molecular methods. Patients were treated with aminoglycoside (streptomycin, gentamicin, amikacin) or quinolone (ciprofloxacin, levofloxacin) antibiotics. Treatment failure was observed in five patients, due to the delay in initiating treatment. Comparison of characteristics and risk factors for tularemia cases versus controls yielded age and contact with rodent excreta at home as potential risk factors (p= 0.001 and 0.002, respectively). The epidemic was controlled after cleaning the tank collecting spring water and chlorination of the water. Tularemia which is an emerging disease in Turkey is spreading to non-endemic regions and represent a significant threat for public health. PMID- 21644067 TI - [Investigation of reduced vancomycin susceptibility in methicillin-resistant staphylococci]. AB - The first Staphylococcus aureus strain with reduced susceptibility to vancomycin was reported from Japan in 1996, and since then an increasing numbers of cases had been reported from various countries. Along with the unfeasibility in the identification of these strains with routine laboratory methods, the use of glycopeptid antibiotics in infections due to these strains may result in therapeutic failure. The aim of this study was to investigate the prevalence of vancomycin intermediate staphylococcus (VIS) and heterogenous VIS (hVIS) strains with the use of agar screening, macro E-test, and population analysis profile (PAP-UC; population analysis profile-area under the curve) methods. A total of 148 methicillin-resistant staphylococcus strains isolated from different clinical samples (48 tracheal aspirate, 48 blood, 39 wound swabs, eight urine, two cerebrospinal fluid, two pleural fluid, one catheter tip sample) between November 2007 and May 2009, were included in the study. Of the isolates 107 were identified as S.aureus and 41 were coagulase-negative staphylococci (CNS; 23 Staphylococcus epidermidis, six Staphylococcus haemolyticus, five Staphylococcus chromogenes, three Staphylococcus hominis and four others) by API Staph kit (bioMerieux, USA). Methicillin resistance has been determined by standard disk diffusion method with oxacillin (1 ug) and cefoxitin (30 ug) disks, according to "Clinical and Laboratory Standarts Institute (CLSI)" guidelines. For the identification of VIS and hVIS strains, brain-heart infusion agar plates containing 6 ug/ml vancomycin (BHI-V6) were used for screening. The suspected VISA/hVISA strains which grew in this agar were further tested by macro E-test and PAP-AUC methods. Total VIS and hVIS rates among the tested isolates, were found as 3.4% (5/148) and 1.4% (2/148), respectively. These rates for CNS strains were 9.8% (4/41) and 2.4% (1/41), and for S.aureus strains were 0.9% (1/107) ve 0.9% (1/107), respectively. In the evaluation of the seven patients who were infected with VISA/hVISA strains, it was detected that all had history of use of glycopeptid antibiotics except one whose history was not reached, and all were hospitalized in intensive care units, except one who had an infected knee prosthesis. Since macro E-test and PAP-AUC methods could not be performed for all of the isolates, there was a probability that our resistance rates did not reflect the real results, nevertheless VIS and hVIS prevalence that we found in our study, seemed to be higher than those data reported previously from our country. In conclusion, since the number of VISA/hVISA strains may increase in time, surveillance for vancomycin resistance in methicillin-resistant staphylococci should be carried out in hospitals periodically. PMID- 21644068 TI - [Comparison of oxacillin, cefoxitin, ceftizoxime, and moxalactam disk diffusion methods for detection of methicillin susceptibility in staphylococci]. AB - Penicillin binding protein 2a/2' (PBP2a/PBP2') which is encoded by the mecA gene, is responsible for the methicillin resistance in staphylococci. Detection of methicillin resistance with phenotypic methods is still a problem especially because of heterogenous expression of mecA gene. Although mecA gene determination by polymerase chain reaction is considered as the gold standard method, molecular tests are not easily applied in all routine laboratories. Thus, for the rapid and accurate diagnosis of MRSA strains, easy and practical phenotypic tests are still required. This study was conducted to compare the oxacillin (OX), cefoxitin (CFX), ceftizoxime (CZX), and moxolactam (MOX) susceptibility testing by disk diffusion method for the detection of methicillin resistance in staphylococci. A total of 247 staphylococci (125 Staphylococcus aureus and 122 coagulase-negative staphylococci; CNS) isolated from various clinical specimens (114 wound and soft tissue materials, 51 urine, 48 blood, 30 respiratory tract, and four other samples) of inpatients and outpatients, were included in this study. PBP2a latex agglutination test was used as the reference method for the recognition of methicillin resistance; four antibiotic disks tested and sensitivity, specificity, positive and negative predictive values (PPV and NPV) were determined for each of them. According to PBP2a latex agglutination test 66 (54.1%) of CNS and 53 (42.4%) of S.aureus isolates were found methicillin- resistant. OX and MOX disks detected 113 (63 CNS and 50 S.aureus) methicillin resistant strain out of 119 PBP2a positive isolates, where CFX and CZX disks detected 110 (60 CNS and 50 S.aureus) of them. Among 128 PBP2a negative isolates, 123 (52 CNS and 71 S.aureus) were detected as susceptible with OX, 127 (55 CNS and 72 S.aureus) with CFX and CZX, 126 (54 CNS and 72 S.aureus) with MOX. According to these results, the sensitivities and specificities of OX, CFX, CZX, and MOX disks were; 95.4% and 92.8%, 90.9% and 98.2%, 90.9% and 98.2%, 95.4% and 96.4%, respectively for CNS and 94.3% and 98.6%, 94.3% and 100%, 94.3% and 100%, 94.3% and 100%, respectively for S.aureus. The difference between sensitivities and specificities of tested antibiotic disks were not found statistically significant. In conclusion, due to the problems in detection of methicillin resistance with phenotypic methods, the use of different mecA gene-inducing antibiotic disks at the same time, and utilization of molecular methods as reference method might be suggested, when a discordance is observed between the antibiotic disks. PMID- 21644069 TI - [Isolation rate of Mycobacterium tuberculosis complex from patients with suspected tuberculosis and identification of the strains with BACTECTM NAP and immunochromatographic TB Ag MPT64 RapidTM Tests]. AB - Tuberculosis (TB) which is still one of the important infectious diseases in the world as well as Turkey, results in high morbidity and mortality. Clinical mycobacteriology laboratories have crucial roles in the identification, typing and susceptibility testing of Mycobacterium tuberculosis. The aims of this study were the investigation of the isolation rate of M.tuberculosis complex (MTC) from the clinical specimens of TB-suspected patients and to compare identification of mycobacteria isolated from solid and/or liquid media by using BACTEC NAP and immunochromatographic TB Ag MPT64 rapid test. A total of 1670 patients who were admitted to outpatients clinics of our hospital and prediagnosed as TB, have been included in the study. All the patients were anti-HIV seronegative. NALC-NaOH method were used for decontamination/ homogenization, and preparations from samples were stained with Erlich-Ziehl-Neelsen method to detect acid-resistant bacilli (ARB) in direct microscopy. All of the samples were inoculated into BACTECTM MGIT-960 (Becton Dickinson, USA) and Lowenstein-Jensen (LJ) media for cultivation and incubated at 37 degrees C for 6-8 weeks. Mycobacteria that were grown in the media have been identified by BACTECTM NAP (Becton Dickinson, USA) and TB Ag MPT64 rapid test (SD Bioline Ag MPT64 RapidTM; Standard Diagnostics, Korea). The culture positivity in the samples of TB-suspected patients was found to be 3.7% (63/1670) with LJ and/or MGIT-960 methods, whereas ARB positivity rate was 1.6% (28/1670). Fifty-three (84%) out of culture positive 63 samples have been identified as MTC by BACTEC NAP test, while 61 (97%) were found as MTC by TB MPT64 test. Considering BACTEC NAP test as the reference method, TB MPT64 test identified all the MTC strains correctly (sensitivity: 100%), however the false positivity rate was estimated as 12.7% (specificity: 87%). Of 53 MTC positive samples, 36 were sputum, four were bronchoalveolar lavage, four were urine, three were gastric fluid, three were pleural fluid, and one of each were abscess, peritoneal fluid and cerebrospinal fluid samples. ARB positivity rate was detected as 41.5% (22/53) among MTC culture positive samples. Of the patients who were infected with MTC, 72% (38/53) were male and 98% (52/53) were adults (age range: 20-85 years). Our data indicating 3.1% (53/1670) isolation rate of MTC from TB-suspected patients in our region were in concordance with the other results reported from Turkey. In conclusion, immunochromatographic TB Ag MPT64 test which seemed to be useful for the rapid identification of mycobacteria grown on solid and/or liquid, was practical to perform and had high sensitivity, however further larger-scaled studies are needed to support our data in our country. PMID- 21644070 TI - [Comparative evaluation of e-test and disk diffusion methods for susceptibility testing of Nocardia species]. AB - Variations in antimicrobial susceptibility among different Nocardia species limit the options for therapy. It is very difficult to perform antimicrobial susceptibility testing of these bacteria due to their slow growth rate and problems in inoculum preparation. The aim of this study was to compare E-test and disk diffusion methods for the determination of antimicrobial susceptibilities of Nocardia isolates. Since E-test is considered as 90% consistent with the gold standard microdilution method recommended by Clinical and Laboratory Standards Institute (CLSI), it was chosen for comparison with disk diffusion and in order to determine the use of disk diffusion in routine practice. A total of 21 Nocardia strains isolated from clinical specimens (12 lung, 7 brain and 2 skin/soft tissue samples) were included in the study. Six of the isolates were identified as N.asteroides, six were N.farcinica, five were N.cyriacigeorgica and four were Nocardia spp. By conventional methods. Susceptibilities of strains to ampicillin, ampicillin-sulbactam, amoxicillin-clavulanic acid, ceftazidime, sefepime, imipenem, gentamicin, erythromycin, levofloxacin, moxifloxacin, trimethoprim- sulfamethoxazole, piperacillin-tazobactam, tigecycline, and linezolid were investigated by using E-test and/or disk diffusion methods. The results were interpreted according to the CLSI breakpoints for Staphylococcus spp. All of the strains were found to be resistant to ceftazidime, piperacillin tazobactam and ampicillin, however susceptible to levofloxacin, moxifloxacin, trimethoprim-sulfamethoxazole tigecycline, and linezolid. The concordance between the methods in terms of susceptibility testing were 100% for ampicillin, ceftazidime, imipenem, gentamicin and linezolid; 85.7% for erythromycin, 76.2% for sefepime, 73.7% for moxifloxacin, 71.4% for piperacillin-tazobactam, 70% for ampicillin-sulbactam and 46.2% for amoxicillin- clavulanic acid. In conclusion, the therapy must be planned according to the results of antimicrobial susceptibility testing. Disk diffusion is not a reliable method due to the high rates of very major errors. E-test would be an alternative method being practical and easily evaluated, especially in routine laboratories in which the reference method could not be performed. PMID- 21644071 TI - [Investigation of BK and JC virus DNA positivities by real-time polymerase chain reaction in the clinical samples of patients with high risk]. AB - Human polyomaviruses, namely BK (BKV) and JC (JCV) viruses are small DNA viruses that cause latent infections worldwide. Primary infections are usually acquired in the early periods of life and are generally asymptomatic. However BKV/JCV infections may cause severe clinical conditions in immunosuppressive patients such as bone marrow and solid organ transplantation or cancer patients. The aim of this retrospective study was to investigate the presence of BKV and JCV nucleic acids by real-time polymerase chain reaction (RT-PCR) in the clinical samples of patients with high risk. A total of 268 (62 blood, 206 urine) samples obtained from 115 immunocompromised patients hospitalized in Gazi University Hospital between July 2007 to January 2009, were included to the study. Viral nucleic acids were extracted from the samples with High Pure PCR Template Preparation Kit (Roche, Germany). By using amplification mix (TIB Molbiol GmbH, Germany) that included primers targeting 174 (JCV) and 219 (BKV) base pair fragments of the small t antigen, and hybridization probes (Roche, Germany), nucleic acids were amplified with LightCycler (Roche Applied Science, Germany) system. As a result, total polyomavirus DNA positivity rate was found as 33.2% (89/268). When BKV and JCV DNA positivities were evaluated according to the samples, 25.2% (53/206) of urine samples yielded positive results for BKV, 14.5% (30/206) for JCV and 2.4% (5/206) for both BKV and JCV. Only one of the blood samples (1/62; 1.6%) were found positive by means of BKV DNA, while none of the blood samples were positive for JCV DNA. The distribution of BKV and JCV DNA positivity rates according to the inpatient clinics were as follows, respectively; 24.3% and 9.5% for pediatric nephrology, 9.6% and 8.2% for renal transplantation unit, 13.5% and 18.9% for adult nephrology, 30.8% and 15.4% for bone marrow transplantation unit, 22.9% and 8.6% for pediatric clinics. In samples from pediatric hematology patients, BKV positivity was 36.4% (4/11), while there were no JCV positivity. However, in hematology patients, while JCV was positive in one of the three samples, no BKV positivity was detected.BKV was seen in three of six samples obtained from patients in the intensive care unit. JCV was positive in both of the two samples obtained from patients in pediatric endocrinology. The only patient that had BKV DNA in blood sample was a renal transplant patient. BKV + JCV DNAs were positive together in only five (1.9%) of the urine samples. In 24% (22/89) of the samples, BKV DNA was found >= 107 copies/ml, in 2.2% (2/89) JCV DNA was >= 107 copies/ml, whereas in 2.2% (2/89) of samples both BKV and JCV DNA was >= 107 copies/ml. All of those samples with high DNA levels were urine. The data of this study led to the establishment of a collaborative algorithm between the laboratory and clinics in our hospital for the diagnosis and follow-up of the patients in terms of BKV/JCV infections. In conclusion, since BKV/JCV reactivations and infections are crutial in immunosuppressive patients, especially medical centers specialized in bone marrow and renal transplantation, diagnostic and monitoring procedures related to those infections should be programmed. PMID- 21644072 TI - [Investigation of cytomegalovirus positivity in the peripheral blood samples of risky patients by shell-vial cell culture, antigenemia test and real-time polymerase chain reaction]. AB - Cytomegalovirus (CMV) infections in immunocompromised patients and congenital infections in infants have high morbidity and mortality while it may lead to asymptomatic infections in immunocompetent subjects. Serological tests, culture methods, antigenemia tests and molecular methods are applied in the diagnosis of CMV infection. The aim of this study was to investigate the presence of CMV in peripheral blood samples of patients who were at risk for CMV disease by shell vial cell culture, antigenemia test and real-time polymerase chain reaction (RT PCR) methods. A total of 141 blood specimens obtained from 91 patients (33 female, 58 male) with suspected CMV disease were included to the study. Five of the patients were newborns and the others aged between 17-79 years old were bone morrow (n= 81), kidney (n= 4) and liver (n= 1) transplantation patients. Shell vial (Vircell, Spain) cell culture method was applied for CMV isolation from the samples, while the detection of pp65 antigen in blood leukocytes was investigated by indirect immunofluorescence method (CINAkit Argene, Biosoft, France). The presence of CMV DNA in plasma samples was detected by RT-PCR (CMV QNP 2.0 kit; Fluorion, Iontek, Turkey) method. CMV was found positive in 72 (51%) of 141 samples by shell vial, 82 (58.2%) by antigenemia test and 49 (34.8%) by RT-PCR. Considering cell culture as the gold standard, the sensitivity and specificity of antigenemia test were calculated as 81.9% and 66.6%, respectively; and for PCR those rates were 43% and 73.9%, respectively. In addition DNA sequencing (ABI Prism 310 Genetic Analyzer; Perkin Elmer, USA) was performed for the samples of randomly selected three patients out of 15, who were yielded positive results with cell culture and antigenemia tests but negative CMV DNA by RT-PCR. In this analysis CMV DNA was found positive in three of the samples that were found negative by RT-PCR in spite of CMV isolation and positive antigenemia. DNA sequencing of those samples revealed multiple mutations in the probe binding region (gB) of CMV QNP 2.0 kit. It was concluded that for the detection of CMV viremia and viral load in patients under risk for CMV disease, antigenemia and PCR based methods could be applied, however, negative results obtained by PCR targeting CMV gB gene, should remind the possible presence of mutations in the related site and the results should be confirmed by sequence analysis. PMID- 21644073 TI - [Evaluation of serum IgG, IgA and IgM levels as indicators of hepatic fibrosis in patients with chronic hepatitis C infection]. AB - The prediction of development of hepatic fibrosis is of crucial importance in terms of disease monitorization and treatment follow-up of patients with chronic hepatitis C virus (HCV) infections. Liver biopsy which is an invasive and complicated method, still remains as the gold standard method for the diagnosis of liver fibrosis. Recently, non-invasive diagnostic tests to determine the biological markers of liver fibrosis have been developed as a possible alternative to liver biopsy. The aim of this study was to evaluate the levels of serum IgG, IgA and IgM antibodies as possible indicators for hepatic fibrosis among patients with chronic HCV infection. A total of 57 patients (35 female, 22 male; mean age: 51 +/- 8.9 years) who were followed-up between January 2007 November 2008, were enrolled in the study. All of the patients were positive for serum anti-HCV and HCV-RNA, while none of them were under antiviral therapy for the last six months. The patients were hospitalized for liver biopsy and biopsy samples were evaluated according to Modified Knodell Histological Activity Index. Forty-nine patients with no liver fibrosis or low to moderate fibrosis were classified as Group 1 (stage 0, 1, 2, 3) and eight patients with high to severe fibrosis were classified as Group 2 (stage 4, 5, 6). Serum IgG, IgA and IgM levels of the patients were determined by a commercial immunonephelometric method (Dade Behring, Germany). Increased antibody levels were detected in a total of 61.4% (35/57) of patients, of which 28 (49.1%) yielded high IgG, 5 (8.8%) yielded high IgM and 2 (3.5%) yielded high IgA levels. The mean IgG levels of patients in Group 1 and 2 were 16.3 +/- 4.6 and 21.8 +/- 5.2 g/L; mean IgM levels were 1.3 +/ 0.6 and 1.6 +/- 0.8, and median IgA levels were 2.0 (0.5-5.3) and 3.3 (1.3- 4.3) g/L, respectively. IgG and IgA levels of patients from Group 2 were found significantly higher than those patients from Group 1 (p= 0.003, p= 0.03, respectively), however there was no significant difference between the groups with respect to serum IgM levels (p= 0.311). When the patient groups were also evaluated in terms of other parameters, no statistically significant differences were detected for ALT, AST, HCV-RNA levels and mean ages (p= 0.95, p= 0.21, p= 0.73, p= 0.10, respectively), however, anti-HCV levels were found significantly higher in Group 2 (p= 0.043). The data of this study indicated a significant relationship between the levels of serum IgG, IgA and the severity of hepatic fibrosis among patients with chronic HCV infection. It was concluded that high serum IgG and IgA levels may be helpful indicators together with the other non invasive markers for the prediction of liver fibrosis in case when liver biopsy could not be performed. PMID- 21644074 TI - [Investigation of antifungal susceptibilities and some virulence factors of Candida strains isolated from blood cultures and genotyping by RAPD-PCR]. AB - Candidemia is a serious clinical picture with a rather high mortality. Early diagnosis and appropriate treatment is crucial in this picture especially in immunocompromised cases. The aims of this retrospective study were to investigate the antifungal susceptibility patterns and to detect the presence of phospholipase, esterase and biofilm production which are excepted as virulence factors of Candida spp. strains and to evaluate the clonal relationships between isolates. A total of 46 Candida spp. Strains isolated from blood cultures of patients of whom eight were newborn and 38 were adults, between the period of February 2005 to July 2010, were included in the study. Of the isolates 17 were identified as C.albicans, 18 were C.parapsilosis, five were C.glabrata, four were C.tropicalis, one was C.guilliermondii and one was C.krusei. Antifungal susceptibility tests were performed by using "Sensititre Yeast One (Trek Diagnostic Systems, USA)" commercial kit. Esterase activity was detected in Tween 80 medium; phospholipase activity in yolk egg agar and biofilm formation was investigated by microplate assay. Strain genotyping was performed by RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction) by using OPE-03, OPE 18, AP50-1, Cnd-3 and Cnd-4 primers. All strains were found to be susceptible to amphotericin B, voriconazole, posaconazole, and caspofungin. C.krusei strain was defined as resistant (intrinsically) to fluconazole. All strains of C.albicans, C.parapsilosis, C.glabrata, and C.tropicalis were found to be susceptible to fluconazole. Three of five C.glabrata strains were resistant to itraconazole, while the other strains were found to be susceptible. All of the C.albicans strains had phospholipase and esterase activity, however none were biofilm producing isolates. In contrast all of the C.parapsilosis strains were negative for phospholipase and esterase activity, however all were positive for biofilm formation. Phospholipase activity has not been detected in non-albicans strains; esterase activity were found positive in all of the C.tropicalis strains, while biofilm formation was detected in three C.tropicalis, one C.glabrata and one C.krusei isolates. The results of genotyping demonstrated that C.albicans strains displayed 5-8 different patterns and C. Parapsilosis strains 2-3 patterns with the use of five primers. Among C.parapsilosis strains, 14 were found identical (with the use of all the primers forming a single pattern (pattern A). In conclusion, the Candida spp. Isolated from blood samples were highly susceptible to the tested antifungals, and C.albicans strains had high phospholipase and esterase activity, while C.parapsilosis strains had high rate of positivity for biofilm formation. The predominant pattern amongst C.parapsilosis strains was thought to be related to exogenous dissemination. PMID- 21644075 TI - [Evaluation of risk factors in patients with candiduria]. AB - Urinary system infections are usually bacterial, however, fungal etiology, particularly Candida spp. are encountered in about 10% of these infections. C.albicans is still the most frequently isolated species in candiduria. This study was aimed to identify the risk factors of candiduria and to determine species distribution of Candida which cause candiduria in hospitalized patients. The study was carried out in a total of 93 hospitalized patients (68 female, 25 male; age range: 17-84 yrs, mean age: 59.5 +/- 1.7 yrs) of which 50 presented with candiduria (case group) and 43 with bacteriuria (control group), between January 2009 to December 2009. The most frequently isolated species was C.albicans (n= 32; 64%), followed by C.glabrata (n= 13; 26%), C.tropicalis (n= 4; 8%) and C.krusei (n= 1; 2%). All of the isolates except one, were found susceptible to fluconazole and voriconazole by E-test (AB Biodisk, Sweden), however, C.krusei isolate was resistant to fluconazole and susceptible to voriconazole. The mean hospitalization period and the period of stay in intensive care unit (ICU) of the case group (9.56 +/- 9.09 and 4.12 +/- 7.05 days, respectively) were found statistically significant compared to control group (4.42 +/- 3.71 and 0.53 +/- 1.78, respectively) (p< 0.005). Nosocomial origin of infection was higher in control group (n= 45, 90%) than the case group (n= 30, 69.8%), (p= 0.014). The rate of antibiotic use prior to candiduria in the case group was detected significantly higher (n= 43; 86%) than the controls (n= 14; 32.6%) (p= 0.000). The most frequently used antibiotic prior to candiduria/bacteriuria was the quinolone group of agents both in case and control groups (42% and 21%, respectively). The other risk factors for candiduria found to be higher in the case group than the controls were as follows; presence of urinary system intervention (32% and 0, respectively; p= 0.000), catheter use (76% and 46.5%, respectively; p= 0.003) and immunosuppression history (24% and 9.3%, respectively; p= 0.041). However, there was no significant relationship between candiduria and history of surgical intervention, diabetes mellitus and renal failure (p> 0.05). In conclusion, rate of candiduria might be reduced by judicious antibiotic use, by implementation of guidelines for urinary catheter use, care and maintenance, and shortening the duration of ICU and hospital stay. PMID- 21644076 TI - [Evaluation of the significance of molecular methods in the diagnosis of invasive fungal infections: comparison with conventional methods]. AB - Direct microscopy and culture methods are still valuable standard conventional methods for the diagnosis of infections caused by true or opportunistic fungal pathogens, especially in high risk patients. However, some of the problems concerning the application and interpretation of those methods, indicate a need for more rapid, practical and reliable tests with high sensitivity and specificity. This study was conducted to compare the results obtained by molecular methods with the results of conventional methods performed simultaneously for the detection and identification of causative fungi in clinical samples. Clinical samples [24 bronchoalveolar lavage (BAL); 14 blood; 5 peritoneal, 4 pleural and 1 pericardial fluids; 1 cerebrospinal fluid (CSF), 1 urine] from 50 immunosuppressed patients were included in the study. All of the samples were cultivated on Sabouraud dextrose and brain-heart infusion agar media and incubated at 30 degrees C and 37 degrees C for 30 days. Samples other than blood were stained with 10-15% KOH + calcofluor white and examined by direct microscopy. Conventional identification of the isolates were performed by using basic morphological and biochemical characteristics. The isolation of fungal DNAs for polymerase chain reaction (PCR) was achieved by classical phenol-chloroform isoamylalcohol procedure (9-10 hours) and commercial DNA extraction kit (6-7 hours) and general and species-specific primers (multiplex) from ITS1, ITS2, ITS3, ITS4, 5.8S rDNA and 28S rDNA regions were chosen for amplification. In PCR results, 550 base-paired (bp) bands obtained with universal primers were evaluated as fungal DNA positivity, and 273 bp, 320 bp, 423 bp, 357 bp, 136 bp and 385 bp bands with species-specific primers were evaluated as Candida albicans, Candida parapsilosis, Candida glabrata, Candida tropicalis, Cryptococcus neoformans and Aspergillus fumigatus positivities, respectively. Seventeen (34%) of the 50 samples yielded fungal growth on culture (C.albicans in 12 BAL, 3 blood, 1 urine sample, and C.parapsilosis in 1 urine), while seven BAL out of 36 (19.4%) non-blood samples gave positive result by direct microscopy. Of the samples 27 (54%) were found positive by PCR. All of the 17 culture positive samples were also found PCR positive, and all of the 23 culture negative samples were also found PCR negative. However, fungal DNAs were detected by PCR in 10 of the samples (5 BAL, 4 peritoneal fluids, 1 CSF) which were negative by direct microscopy and culture methods. These fungi were identified as C.albicans (n= 8), C.parapsilosis (n= 1, from peritonal fluid) and C.neoformans (n= 1, from CSF) by multiplex PCR. No samples yielded PCR negative, culture positive result. All of those 10 PCR positive, culture negative samples belonged to patients who were under antifungal treatment. The detection of C.neoformans DNA from CSF sample of a patient with suspected cryptococcosis only with PCR provided the chance for rapid therapy. In statistical evaluation, the concordance between culture and PCR methods were found significantly high (k= 0.61; p< 0.001), whereas it was minimal (k= 0.24; p< 0.001) between direct microscopy and PCR. When considering culture as the reference method, the sensitivity and specificity of PCR were estimated as 100% and 69.7%, respectively. In addition, multiplex PCR was as successful as culture and conventional identification methods in the identification of all fungal species. As a result, without disregarding conventional methods, use of PCR might be recommended for the identification of fungal species on the basis of clinical status of the patient and conditions of the laboratory. PMID- 21644077 TI - [Experimental oral candidiasis in healthy and immunocompromised BALB/c mice]. AB - Oral candidiasis which is the most common type of Candida infections affecting humans, is most frequently caused by C.albicans. Immune response of the host, as well as a variety of virulence factors of the causative agent, play important roles in the development of Candida infections. The colonization rate of Candida in the oral cavity of healthy individuals, is between 25-30%, however, this rate is reported to be increased in immunosuppressive subjects. In our study, we established an oral candidiasis model with C.albicans in healthy and experimentally immunocompromised mice and aimed to compare Candida colonization rates and histopathological changes occurred in the tongue and esophagus tissues of the animal groups. A total of 21 BALB/c mice were grouped as control (Group 1; n= 7), healthy (Group 2; n= 7) and immunocompromised (Group 3; n= 7) groups. Immunosuppression in mice was performed by subcutaneous injection of prednisolone. For experimental oral candidiasis, cotton swab impregnated with C.albicans strains which did not have acid proteinase and phospholipase enzyme activity, no biofilm production, and sensitive to fluconazole and amphotericin B, were used. In the control group, physiological saline solution was used instead of C.albicans strain. In the forth day of experimental oral candidiasis model swab samples taken from the dorsal tongue surface of mice were evaluated by quantitative cultivation method. No yeast colonies were detected in Group 1 while more significant number of yeast colonies were observed in Group 3 compared to Group 2 (p= 0.002). Tongue and esophagus tissues of mice were stained with hematoxylin-eosin and periodic acid schiff staining and evaluated in terms of inflammatory response, abscess formation, vascular congestion, vasodilation and for the presence of yeast and hyphae. When the inflammation in esophagus was considered, statistically significant difference was determined between group 1 and group 3 (p= 0.023), however, no difference was detected between group 2 and 3 (p= 0.107). The level of inflammation in tongue tissue exhibited no difference between groups 2 and 3 (p= 0.317) while the difference was significant when these groups were compared to the control group (p= 0.00, p= 0.002, respectively). Similarly, the level of congestion in tongue tissue exhibited no difference between groups 2 and 3, however, the difference was significant when compared to the control group. To enlighten the relation between host immune status and oral candidiasis caused by C. albicans, further larger-scale studies also concerning the various virulence factors of the infectious agent, should be conducted by the use of experimental animal models which may successfully guide us in this regard. PMID- 21644078 TI - [Investigation of interleukin-10, tumor necrosis factor-alpha and interferon gamma expression in experimental model of pulmonary aspergillosis]. AB - Pulmonary aspergillosis which is an important opportunistic infection in neutropenic patients, is usually caused by Aspergillus fumigatus. Since the pathogenesis of disease is not well understood, the main proposed mechanism is thought to be cell-mediated immunity and cytokine response. The aim of this study was to investigate the local production of cytokines in the lung tissues of rats with experimentally developed aspergillosis, by reverse transcriptase-polymerase chain reaction (RT-PCR). A total of 33 Wistar albino type rats were included in the study with the consent of Experimental Animal Ethics Committee. Twenty-five of the rats were infected with A.fumigatus by intratracheal way, while 8 animals were used as controls. The presence of A.fumigatus in the lung tissues of infected rats was confirmed with the use of quantitative culture and histologic staining methods. RNA isolation from the lung tissue samples of both groups were performed by a commercial kit (Qiagen, Germany). After obtaining complementary DNAs from the genomic RNAs, in-house qualitative and quantitative (real-time) PCR methods were used to amplify the target regions for interleukin-10 (IL-10), tumor necrosis factor-alpha (TNF-?) and interferon-gamma (IFN-?) by using specific primers (Tib Molbiol, Germany). Mean mRNA levels achieved by real-time PCR for IL 10, TNF-? and IFN-? in aspergillosis group were 6.5 x 106 copies/ml, 7.9 x 105 copies/ml and 2.2 x 103 copies/ml, respectively, while those values in control group were 4.3 x 102 copies/ml, 5.6 x 103 copies/ml and 1.3 x 102 copies/ml, respectively. Our data indicated that rat model of aspergillosis was associated with significantly increased expression of mRNA encoding IL-10 and TNF-? than controls (p< 0.05), however there was no statistically significant difference between the groups with respect to IFN-? expression (p= 0.53). In conclusion, the production of proinflammatory cytokines which mediate the influx of phagocytic cells might account for the localization of Aspergillus infection to the upper respiratory tract. The up-regulation of the expression of the immunomodulatory cytokine TNF-? and IL-10 in lung tissue from infected rats might be important to limit the extent of local tissue destruction, but might also account for the fact that infected rats are generally unable to clear the infection spontaneously. PMID- 21644079 TI - [Investigation of occult hepatitis B in HIV infected patients]. AB - Due to their shared transmission route, hepatitis B virus (HBV) or hepatitis C virus (HCV) co-infections can be observed in human immunodeficiency virus (HIV) infected cases and are associated with more severe clinical courses. The detection of HBV DNA despite HBV surface antigen (HBsAg) seronegativity is defined as occult HBV infections. According to the current seroepidemiological data, Turkey is classified as an intermediate HBV, low HIV endemic region. Occult HBV infections have previously been reported from Turkey but has not been investigated previously in HIV infected cohorts. The aim of this study was to identify occult HBV infections in HIV-infected persons. Twenty-eight HIV-positive cases followed-up at Hacettepe University Hospital, Infectious Diseases Unit were included in the study after informed consent. For the detection of HBsAg, anti HBs and anti-HCV, commercial ELISA tests (Architect System, Abbott Diagnostics, USA) were employed. Absolute CD4+ and CD8+ T-cell counts were determined via flow cytometry. HIV viral load was calculated via COBAS TaqMan HIV-1 Real-time PCR (Roche Diagnostics, USA) and the presence of HBV DNA was evaluated via COBAS TaqMan HBV Real-time PCR (Roche Diagnostics, USA), in addition to a nested PCR assay targeting HBV S gene. The mean age of the study group was 43.2 (range between 27-65) years, 64.3% (18/28) of them were males and the mean duration of HIV infection was 4.2 (2-11) years. Mean CD4+ ve CD8+ T-cell counts were 414 +/- 267 cells/mm3 and 854 +/- 293 cells/mm3, respectively. Twenty-six (92.8%) cases were under highly-active anti-retroviral therapy at the time of the study, 88.5% of which included HBV-active drugs (lamivudine or tenofovir). HIV RNA were found negative in 11 (39.3%) patients, of those nine (81.8%) were the cases who treated with HBV-active antiretroviral therapy. HBsAg were negative in all of the 28 patients, while the positivity rates of anti-HBs and anti-HCV were 39.3% (11/28) and 3.6% (1/28), respectively. All samples were negative for HBV DNA via the commercial real-time PCR and in-house nested PCR assays. The absence of occult HBV in the study group may indicate the absence of occult HBV or suppression of viral replication due to the anti-retroviral therapy. In conclusion, further large-scale studies are required to fully understand the impact of occult HBV in HIV-infected patients in Turkey. PMID- 21644080 TI - [Prevalence of hepatitis B virus infection in children of HBsAg positive parents]. AB - Hepatitis B virus (HBV) is one of the important causative agents of chronic liver disease in the world. Since HBV transmission is mainly by parenteral way and sexual contact, vertical (from infected mother to newborn) and horizontal (contact with infected subjects) transmission ways play important roles. The aim of this study was to retrospectively evaluate the intrafamilial transmission risk of HBV in our region. A total of 838 subjects (age range: 1-52 years; mean age: 19.5 years) whose parents were HBsAg positive and were admitted to our outpatient clinics in 2009, have been included to the study. Of them 389 (46.4%) had HBsAg positive mothers, 404 (48.2%) had HBsAg positive fathers and 45 (5.4%) cases had both parents as HBsAg positive. The rate of chronically [corrected] infected subjects (HBsAg positive, anti-HBs negative) with HBsAg positive parents was 14.4% (121/838), the rate of non-infected susceptible subjects (HBsAg negative, anti-HBs negative) was 38.4% (322/838), and the rate of subjects who developed immunity (HBsAg negative, anti-HBs positive) was 5.3% (44/838). HBsAg positivity rate was found to be high in subjects whose mothers were HBsAg positive (25.2%; 98/389), than those whose fathers were HBsAg positive (2.5%; 10/404) and the difference was found statistically significant (p= 0.0001). Similarly the rate of HBV infection (HBsAg positive or anti-HBs positive subjects) among the offsprings whose mothers were HBsAg positive was determined to be higher (30.3%) than those whose fathers were HBsAg positive (8.2%) (p< 0.001). According to these data, the risk of HBV transmission was estimated to be three fold higher in subjects whose mothers were HBsAg positive than those whose fathers were HBsAg positive (OR= 2.96, 95% CI 2.109-4.156). In our study it was also detected that the rate of HBsAg positivity increased with age, being 10.3% (6/58) in children below 10 years old and increasing to 33% (29/88) in 11-20 years, 47% (31/66) in 21-30 years and 65% (26/40) in > 30 years age groups. There was no significant difference between consecutive age groups, however the difference between < 10 years and > 30 years age groups was found statistically significant (p= 0.0001). In addition having HBsAg positive mother or father did not affect HBV infection rate at 0-10 years age group, however, having HBsAg positive mother significantly increased the infection rate more than HBsAg positive father in 11-20 years and over 21 years age groups (p= 0.001 and p= 0.0001, respectively). In conclusion, the transmission rate of HBV infection among the family members was found high in our region and especially mothers played an important role for the transmission of the virus to their offsprings. PMID- 21644081 TI - [Chronic renal failure: unexpected late sequela of pulmonary tuberculosis after 30 years]. AB - Tuberculosis-related chronic granulomatous tubulointerstitial nephritis (GTN) and chronic renal dysfunction as a consequence of GTN is a rarely seen clinical condition, with a few case reports in the literature. In this report, a case with end stage renal failure as an unexpected late extrapulmonary sequela of tuberculosis has been presented. A 60 years old female patient was admitted to hospital with the complaints of fever, malaise and nausea. Her history revealed that she had pulmonary tuberculosis 30 years ago and received antituberculosis therapy for nine months. The laboratory results on admission were as follows: blood urea nitrogen 90 mg/dl, serum creatinine 9 mg/dl, sodium 116 mEq/L, potassium 6.6 mEq/L, albumine 2.9 g/dl, hemoglobin, 8.4 g/dl, white blood cell count 10.800/mm3, C-reactive protein 187 mg/L and erythrocyte sedimentation rate 110 mm/hour. Urinalysis showed 8.1 g/L protein, 10-12 leukocytes, 1-2 erythrocytes, while 24-hours urinalysis yielded proteinuria with 8 ml/minutes creatinine clearance value. Urine and blood cultures of the patient revealed neither bacteria or mycobacteria. PPD skin test was negative. Acid-resistant bacilli (ARB) were not detected in sequential urine samples obtained on three consecutive days. Since sputum samples could not be obtained, diagnostic procedures for sputum were not performed. Abdomen ultrasonography yielded bilateral edema and grade II echogenity in kidneys. Computed tomography of the chest showed bilateral pulmonary nodules, chronic sequela lesions, pleural scarring and calcifications, as well as minimal interstitial infiltrate. Transthoracic lung biopsy showed chronic inflammation and fibrosis, while amyloid was negative. Renal biopsy showed GTN with central caseified necrosis and granulomas, multinuclear giant cells, tubular atrophy and interstitial fibrosis. Amyloid was negative and ARB were not detected in renal biopsy sample. Definitive diagnosis was achieved by the demonstration of Mycobacterium tuberculosis nucleic acid in kidney biopsy by polymerase chain reaction (PCR). Antituberculosis therapy was not initiated since there were no signs of active tuberculosis. The patient became clinically stable following dialysis and was discharged, however, she has been undergoing hemodialysis three times a week. The aim of this case presentation was to emphasize that renal tuberculosis should be considered in the differential diagnosis of patients with end stage renal failure, especially in countries like Turkey where tuberculosis incidence is high. PMID- 21644082 TI - [Current approaches to explain the virulence of Acinetobacter baumannii]. AB - Acinetobacter baumannii which is one of the most frequent nosocomial pathogens, has drawn attention in the last years owing to multi-drug resistant strains. A.baumannii may give rise to nosocomial epidemics especially in intensive care units and may lead to treatment failure due to its increasing antimicrobial resistance. These gram-negative non-fermentative coccobacilli may be encountered also in community associated infections. However, they are frequently isolated in pneumonia, urinary tract infection, bacteremia, meningitis and wound infections that develop in patients hospitalized for serious diseases. Although detailed data about the epidemiology and antimicrobial resistance patterns related to this bacteria exist, relatively limited data is present about the virulence factors and environmental physiology of A.baumannii. The role of some bacterial virulence factors in the pathogenesis of Acinetobacter infections have been enlightened by recent investigations. Among these virulence factors, production of extracellular enzymes with lipolytic and cytolytic activities, outer membrane protein (AbOmpA) with apoptotic effects on epithelial cells, adhesion molecules (fimbria and AbOmpA) that function during attachment to epithelial cells, K1 type capsular structure, type-1 pili and AbOmpA induced biofilm formation, siderophore (acinetobactin) or hemin mediated iron acquisition mechanisms, quorum sensing system that functions by the help of N-acyl homoserine lacton signal molecules and cellular components that enable Acinetobacter species to live under inappropriate environmental conditions like dryness, low temperature, restricted nutritional elements, can be counted. New information about the virulence factors will help better understanding of the adaptive response of A.baumannii in the host setting. This review is focused on the current information about the virulence factors of of A.baumannii. PMID- 21644083 TI - [Confirmation of West Nile virus seroreactivity in central nervous system infections of unknown etiology from Ankara Province, Central Anatolia, Turkey]. AB - West Nile virus (WNV) infections may trigger febrile conditions and/or neuroinvasive disease in a portion of the exposed individuals. Serosurveillance data from various regions of Turkey indicate WNV activity. The aim of this study was to confirm the antibody specificity of the serum samples via virus neutralization assay, previously reported to be reactive for WNV IgM. The samples originated from two individuals with the preliminary diagnosis of aseptic meningitis/encephalitis of unknown etiology in 2009 and had been classified as probable WNV infections. Cerebrospinal fluid and sera samples of these patients had been evaluated as negative for WNV RNA and IgG antibodies. Only one serum sample could be included in the neutralization assay due to the limited amounts in the current investigation. The sample was observed as positive in dilutions of 1/20 and 1/40, thus confirming the diagnosis of WNV-related central nervous system infection in a 62 year-old female patient from Ankara, Central Anatolia, Turkey. PMID- 21644084 TI - Evaluation of the effects of losartan on a random pattern skin flap model in rats. AB - BACKGROUND: Losartan, a drug with antiinflammatory properties, has been shown to prevent ischemic injury in various organs. The purpose of the present study was to investigate the effects of losartan on ischemic skin flaps and on flap survival. METHODS: A 3x9 cm dorsal flap was elevated in 24 Sprague-Dawley rats. Rats received the following treatment for seven days: Group I (n=8): 10 mg/kg losartan; Group II (n=8): 40 mg/kg losartan; and Group III (n=8): nonionized water. At the end of the seventh day, the viable flap areas were calculated, and histological analysis was conducted to count cells and capillaries in microscopic fields. RESULTS: Mean flap survival was 61%, 56% and 60% in Groups I, II and III, respectively. Comparison of flap survival between groups did not yield any significant difference (p>0.05). Ischemic zones of the flaps in all groups demonstrated an increase in number of neutrophils, fibroblasts and capillaries (p<0.05), whereas no difference was seen in mast cells. The cell counts in the viable areas of the flaps showed a significant decrease in fibroblasts in the group treated with 40 mg/kg losartan (p<0.05). The number of neutrophils, mast cells and capillaries was not influenced by treatment. CONCLUSION: Losartan does not improve skin flap survival but it has a significant antiproliferative effect on fibroblasts. PMID- 21644085 TI - Effects of spinal immobilization devices on pulmonary function in healthy volunteer individuals. AB - BACKGROUND: We aimed to investigate the effects of spinal immobilization devices on pulmonary functions. METHODS: This study was a cross-over trial in healthy volunteer subjects; 60 volunteers were included. We performed a full spirometry in the supine position, and forced vital capacity (FVC), forced expiratory volume in one second (FEV1) and FEV1/FVC were recorded in all subjects. Then, Philadelphia type cervical collar (Philly) and Kendrick extrication device (KED) were applied to all subjects. We measured FVC, FEV1, FEV1/FVC in all subjects in the supine position at the 5th and 30th minutes after application of cervical collar and KED. After a one-hour relaxation period, Philly and long spinal backboard with straps were applied to all subjects. FVC, FEV1, FEV1/FVC were measured again in all subjects at the 5th and 30th minutes after application of cervical collar and long spinal backboard. RESULTS: After application of KED, baseline levels were compared with levels at the 5th and 30th minutes. Statistically significant decreases were determined in FEV1 (p=0.000) and FVC levels (p=0.000) after application of KED, but there were no significant differences in FEV1/FVC levels. After application of the long spinal backboard, a comparison of baseline levels and levels at the 5th and 30th minutes demonstrated statistically significant decreases in FEV1 (p=0.000) and FVC levels (p=0.000), but no significant difference in FEV1/FVC levels. CONCLUSION: We determined that both KED and long spinal backboard cause a decrease in pulmonary functions. PMID- 21644086 TI - Evaluation of agar films in the prevention of postoperative peritoneal adhesions in an animal model. AB - BACKGROUND: Peritoneal adhesions cause significant long-term postoperative morbidity. This study evaluates the efficacy of agar plates as the physical barrier in reducing adhesion formation after abdominal surgery in an animal model. METHODS: Adhesions were induced, by cecum abrasion, in 20 C57/BL6 mice during a laparotomy procedure. Agar plates were used in 10 mice as the experimental group. At a second operation, 28 days later, the adhesions were graded, in two groups. Data were analyzed by using Student t test. RESULTS: There was no significant difference in weight gain of the two groups during the study period. A comparison of the morphological appearances of the adhesions demonstrated that there was no evident difference between the two groups. There was also no significant difference in the incidence ratio of adhesions or postoperative adhesion scores between the two groups (p value >0.05). CONCLUSION: Despite the hydrogel properties of agar, it was not successful in practice in the reduction of adhesion formation after peritoneal surgery. Since agar is a biological product, it may cause a hyperreactivity induced by the innate immune system in peritoneum. Therefore, agar does not appear to be useful in clinical practice for the reduction of adhesion formation after peritoneal surgery. PMID- 21644087 TI - Paramedic-performed Focused Assessment with Sonography in Trauma (FAST) in the emergency department. AB - BACKGROUND: Our objective was to evaluate the accuracy of paramedic-performed Focused Assessment with Sonography in Trauma (PFAST) for detection of free fluid in patients admitted to the Emergency Department (ED) following trauma. METHODS: After four hours of didactic and four hours of hands-on training, four paramedics prospectively evaluated trauma patients. Our gold standard was the official radiologist reports of ultrasonography and computerized abdominal tomography (CAT). The sensitivity, specificity, positive and negative likelihood ratios, and diagnostic odds ratio of PFAST were calculated and analyzed using SPSS 15.0 with ?2 testing. RESULTS: One hundred and twenty-seven patients were evaluated by the paramedics. Fourteen patients had positive free fluid in the abdomen. Of these, 11 were corroborated by radiology reports and CAT (true positives), and three were found to be negative (false positives). In 113 cases, PFAST was negative for free fluid. Of these, 111 were determined not to have free fluid (true negatives), whereas free fluid was detected by CAT in 2 (false negatives). The sensitivity, specificity, positive and negative likelihood ratios, and diagnostic odds ratio of PFAST were 84.62, 97.37, 32.15, 0.16, and 203.50, respectively. CONCLUSION: Our study shows that paramedics can perform FAST in hospital Eds with a high degree of accuracy. PMID- 21644088 TI - [Should videothorascopic surgery be the first choice in isolated traumatic hemothorax? A prospective randomized controlled study]. AB - BACKGROUND: In this study, patients with hemothorax due to blunt or penetrating thorax trauma and treated by tube thoracostomy were compared with the patients treated by videothoracoscopic surgery (VATS) in order to determine whether VATS can be the first choice in treatment. METHODS: Sixty patients with hemothorax due to trauma were examined prospectively. Thirty patients with isolated hemothorax and treated by tube thoracostomy were classified as Group I (50%), and 30 patients treated by VATS were classified as Group II (50%). Patients were compared according to healing duration, tube thoracostomy duration, hospitalization duration, success rate of treatment, morbidity, and mortality. RESULTS: When Group I and II were compared according to the duration of tube thoracostomy and hospitalization, Group I was found to have longer duration statistically (p=0.001). When the two groups were compared according to morbidity, statistical morbidity was found higher in Group I (p=0.030). CONCLUSION: VATS seems to be better in the treatment of hemothorax when compared with traditional drainage method because of its advantage of direct diagnosis and hemostasis by diagnostic thoracoscopy. If clotted blood is evacuated, empyema and fibrothorax are preventable. In these cases, chest tube duration and hospital length of stay are shorter. Therefore, these patients can be discharged early. PMID- 21644089 TI - Clinical evaluation of a temporary fecal containment device for non-surgical fecal diversion in perineal burns. AB - BACKGROUND: The aim of this study was to evaluate the clinical results of a temporary fecal containment device (Flexi-Seal(r) FMS) in our burn center. METHODS: All patients in whom temporary fecal containment devices were applied for perineal burns between August 2008 and August 2009 in our institution were reviewed. Demographics, etiology of burns, total body surface area (TBSA) burned, intensive care unit (ICU) need, early mortality, and post-application data were obtained from a prospectively designed database. In addition, some variables were investigated as potential risks factors for fecal leakage. RESULTS: The mean age of patients (n=15) was 43.1+/-22.1 years, and 66.7% of the patients were male. The mean %TBSA burned was 40.7+/-16.6. Fecal leakage was seen in 6 patients. Local infection in the perineum was observed in 6 patients, including 4 of the 6 patients with fecal leakage. The mortality rate was 33% (5 deaths). All exitus patients had 50% or more TBSA burned. Electrical burn injury was found as a significant risk factor for fecal leakage in surviving patients (p<0.05). Autologous split-thickness grafting was performed in 8 patients without complication. The mean duration of catheterization was 22.5+/-5.7 days. Except for superficial mucosal erosion in the distal rectum in 2 cases, no complication was observed. The mean hospitalization time was 46.7+/-12.7 days. CONCLUSION: If the safety of these devices is proven in further prospective, high-volume studies, they may reduce the necessity of diverting stoma operation in burn patients. PMID- 21644090 TI - [The association between cellular morphological changes in peripheral blood smear and complications in pediatric burn cases]. AB - BACKGROUND: Mortality and morbidity in burn cases can be reduced with early diagnosis. Many markers are used for early diagnosis of burn complications like sepsis. In this current study, the relationship between numerical/morphologic granulocyte abnormalities and complications was investigated in pediatric burns. It was aimed to introduce histopathologic marker(s) for burn-related complications. METHODS: Thirty-two pediatric burn cases hospitalized between December 2006 and December 2009 were included in the study. A total of 192 complete blood count and peripheral blood smear results were analyzed comparatively. Findings were used to identify any correlation among white blood cell count and peripheral blood smear changes (the appearance of immature granular cells, toxic granulation, purple granules and Dohle bodies) and complications such as bacteriemia, sepsis, wound infections, severe anemia, and graft failure. RESULTS: White blood cell count changes and the appearance of immature granular cells were not suitable for use as a diagnostic marker for complications. Nevertheless, there was a statistically significant correlation between the appearance of toxic granulation, purple granules and Dohle bodies and subsequent complications (p: <0.0001, 0.041, 0.001, respectively). CONCLUSION: Toxic granulation, purple granules and Dohle bodies appear to be helpful in predicting burn-related complications. Therefore, peripheral blood smear is a suitable test for predicting future complications. PMID- 21644091 TI - [An evaluation of dog and cat bites over a five-year period: a sample case from Eskisehir]. AB - BACKGROUND: In this study, it was aimed to evaluate the demographic, medical and forensic aspects of patients who were bitten by cats or dogs and admitted to the Eskisehir Osmangazi University, Education, Training and Research Hospital. METHODS: All the medical records of the last five years inside the automation system of the hospital were investigated. One hundred sixty-seven cases were included according to their ICD-10 diagnostic codes. On statistical evaluation, frequency and chi-square analyses were conducted and SPSS 13.0 program was used. RESULTS: One hundred sixteen patients were bitten by dogs, whereas 46 patients were bitten by cats. Most of the victims were children. 51.9% of the bites were on an upper extremity, and 29.1% were on a lower extremity. Although cat and dog bites demonstrate seasonal changes, summer months were found important, when children were on holidays. Hyperemia, scratch and laceration were the most frequent diagnoses after injury. The length of hospital stay was between 1-22 days and calculated as a mean 7.3+/-5.8 days. CONCLUSION: Dog and cat bites are very well known public health issues in our country. To know animals and their needs and to respect them should be the priority of families to avoid unexpected injury. Families should teach their children to treat dogs and cats with respect, to avoid direct eye contact with the animals and not to annoy them. PMID- 21644092 TI - [Effects of short- and long-segment posterior instrumentation on spinal canal remodeling in thoracolumbar vertebra burst fractures]. AB - BACKGROUND: Spinal canal remodeling results according to Magerl classification and fracture localization after short- and long-segment posterior instrumentation treatment were evaluated in patients with thoracolumbar junction burst fracture. METHODS: Eighty patients were divided into two groups: Group 1: short-segment posterior instrumentation was applied in 36 patients [9F, 27M; Median age: 42.1 (range: 19-65)] and Group 2: long-segment posterior instrumentation was applied in 44 patients [18F, 26M; Median age: 46.3 (range: 18-78)]. Twenty patients had T12, 41 patients had L1 and 19 patients had L2 fracture. According to Magerl classification, 44 patients were A3.1, 19 were A3.2 and 17 were A3.3. In both groups, spinal canal remodeling effectiveness was evaluated postoperatively with respect to all parameters. RESULTS: Median follow-up time was 35.7 months for Group 1 (12-58) and 33.1 months for Group 2 (12-58). In both groups, spinal canal remodeling was statistically significant, but a higher recovery ratio was obtained in Group 2 in comparison to Group 1. According to Magerl classification, in type A3.3 fractures, a more significant remodeling was obtained in Group 2 patients (p=0.005). A significant difference was determined in Group 2 at the T12 level according to fracture localization (p=0.018). CONCLUSION: An adequate spinal canal remodeling is obtained by posterior instrumentation, but in comminuted fractures like Magerl type A3.3, a better remodeling can be obtained by long-segment posterior instrumentation. PMID- 21644093 TI - Pattern of skull fracture in Iran: report of the Iran National Trauma Project. AB - BACKGROUND: We characterize in this report the mechanism and type of skull fracture in urban populations of Iran. METHODS: Data including the general characteristics, mechanism of trauma, abbreviated injury scale, Glasgow coma scale, duration of hospitalization, and outcome of trauma patients registered from 1999 to 2004 were extracted from the Iranian National Trauma Registry database. RESULTS: Of 16,321 registered trauma patients, 1704 cases with skull fracture were found. The most common mechanism of trauma was motor-vehicle crash (MVC) (62.5%) followed by fall (23.8%). The majority of traffic victims were pedestrians (41.6%). Skull fracture was more likely to be seen in men (78.6%), with a mean age of 27.2 +/- 17.7 years. In MVCs, skull base fracture was observed in 51.2% and vault fracture in 48.8% of patients. A significant difference was found in sex distribution between skull base and vault fracture (p=0.002). MVC was the mechanism of injury in 67.4% of skull base fracture cases compared to 58.1% of vault fracture cases (p<0.001). CONCLUSION: Safety legislations and risk specific intervention programs should be improved considerably in Iran. PMID- 21644094 TI - Management of community-based shotgun injuries of the extremities: impact of emergent vascular repair without angiography. AB - BACKGROUND: Shotgun injuries of the extremities create challenging problems for vascular surgeons. In this study, we retrospectively analyzed surgical results without preoperative angiography. METHODS: Forty-nine patients with shotgun wounds who underwent vascular reconstruction in the extremities from 1999 to 2004 were retrospectively reviewed. RESULTS: Vascular reconstruction of the extremities after shotgun injury differs from that following injuries caused by other firearms because of extensive tissue damage. In 19 patients, function of the extremity was unsatisfactory after one year; in 12 of them functional deficit was extreme, which was thought to be the result of nerve injury. After several interventions, 25 of 49 patients are well after one year under a rehabilitation program. CONCLUSION: Based on these results, we favor immediate operative exploration of the extremities in patients with hard signs of vascular trauma, thereby minimizing the ischemic interval, and we recommend angiography only for elective operations. Early fasciotomy should be done without hesitation in patients with long ischemic periods and in those with combined arterial/venous injury. PMID- 21644095 TI - Comparison of two biological internal fixation techniques in the treatment of adult femur shaft fractures (plate-screws and locked intramedullary nail). AB - BACKGROUND: We aimed to compare two (plate-screws and locked intramedullary nail) biological internal fixation techniques in the treatment of adult femur shaft fractures. METHODS: Group I included 28 patients operated with biological internal fixation with plate and screws. Group II consisted of 31 patients operated with locked intramedullary nailing technique. During the follow-up period in Group I, 1 patient died and 3 patients dropped out of the study, so the results of 24 patients were evaluated. During the follow-up period in Group II, 2 patients died and 3 patients dropped out of the study, so the results of 26 patients were evaluated in this group. RESULTS: Age, sex, injury mechanism, fracture type, time to operation, duration of operation, amount of bleeding, fluoroscopy usage time, early and late complications, duration of hospitalization, fracture union time, and functional results of all patients were comparatively evaluated. CONCLUSION: According to our results, no superiority of either technique was demonstrated with respect to fracture union time, complication rate and functional results. Biological internal fixation with plate and screws is an alternative technique to locked intramedullary nailing in patients with multi-trauma or compromised pulmonary or cardiac function, or in complex comminuted or segmented diaphyseal or metaphyseal fractures. PMID- 21644096 TI - [Pediatric head injuries, retrospective analysis of 851 patients: an epidemiological study]. AB - BACKGROUND: The aim of this study was to contribute to the epidemiological database of our country about pediatric head injuries. METHODS: We retrospectively analyzed 851 patients younger than 14 years old with head injury, treated in the Neurosurgery Department of Samsun Mehmet Aydin Education and Research Hospital between January 2003 and June 2008. RESULTS: Five hundred and fifty (65%) patients were male and 301 (35%) were female. Falls (70%) were the most common cause of head injury. According to Glasgow Coma Score (GCS), 74% of patients had mild (13-15), 22% moderate (9-12) and 4% severe (3-8) head injury. Eighty-seven (10.5%) patients were operated. The commonest operations were performed for depressed fractures and epidural hematomas. The mortality rate was 3.8% (33). Common causes of mortality were traffic accidents and falls from heights. CONCLUSION: Other reported studies and this study show that the majority of pediatric head injuries occur as a result of preventable causes. It is important to have local epidemiological studies and data about head injuries in order to prevent these traumas before their occurrence. PMID- 21644097 TI - A rare cause of chronic rectal bleeding in children; solitary rectal ulcer: case report. AB - Solitary rectal ulcer causing lower gastrointestinal bleeding is extremely rare in children. Rare presentation, non-specific symptoms, insufficient experience, and characteristics mimicking other rectal diseases may cause misdiagnosis or delay of diagnosis in some pediatric patients. Here, we report a 10-year-old boy with solitary rectal ulcer diagnosed two years after onset of the symptoms who responded well to the conservative therapy, including high-fiber diet, laxatives, defecation training, and sucralfate enema. PMID- 21644098 TI - Replantation of scalp avulsion following a go-kart accident: a case report. AB - With the development of microsurgery, successfully replanted cases of scalp avulsions have been reported. In spite of previous publications of replantations based on a single artery and vein, it is now accepted that multiple anastomoses increase the success rate. We present herein the case of a successfully replanted avulsion in a child who caught her hair in a go-kart motor belt, and we discuss the mechanism of injury and number of anastomoses. PMID- 21644099 TI - Life-threatening abdominal injury during a soccer game: a rare clinical case. AB - Soccer (football) is a popular sport worldwide and can result in severe abdominal injuries. Nevertheless, the necessity of surgical intervention for abdominal organ injuries has been reported rarely. We report a case who was injured during a soccer game who underwent abdominal surgery. Distal subtotal pancreatectomy, splenectomy, cholecystectomy, and choledochotomy + T-tube drainage were performed. He was discharged on the postoperative seventh day without any complication. PMID- 21644100 TI - Delayed retroperitoneal bleeding causing acute abdominal compartment syndrome: case report. AB - Delayed acute abdominal compartment syndrome (ACS) due to retroperitoneal bleeding is rare. Herein, we report the clinical management of such a rare case. A 46-year-old male who fell from a height of 12 meters was admitted to Al-Ain Hospital. He was hemodynamically stable. His abdomen was soft and not distended. Abdominal computed tomography (CT) was normal on admission. On day 7, the patient tolerated enteral feeding. On day 15, he became suddenly hypotensive. CT of the abdomen showed a large retroperitoneal hematoma compressing the inferior vena cava (IVC) associated with contrast blush indicating active bleeding. The abdomen became distended and tense. The patient developed respiratory failure and severe acidosis, increased airway pressure and reduced urine output. A clinical diagnosis of ACS was made. There was dramatic improvement in the hemodynamic and respiratory function directly after laparotomy. Exploration of the retroperitoneal hematoma showed an actively bleeding ligated ileocolic vessel. The abdomen was temporarily closed using saline IV bags sandwiched between two layers of Steri-Drape. The abdomen was closed primarily on day 6. The patient was discharged home on day 50. Life-threatening delayed retroperitoneal bleeding may occur suddenly two weeks after trauma causing ACS. PMID- 21644101 TI - Post-traumatic cerebral infarction: a rare complication in a pediatric patient after mild head injury. AB - Because the cases of post-traumatic cerebral infarction in children are uncommon, little research has been done on this subject. The case of a 14-month-old child who had cerebral infarction after mild head injury is discussed. He fell from a height of approximately 70 cm 12 hours before. He did not use his left arm after the injury. His parents took him to the public hospital, where he was investigated in terms of mechanical complication and was observed for six hours, but no computed tomography (CT) scan was performed at that time. The patient was then presented to our department by his parents. He was not able to walk and was sitting. His neurological examination revealed right hemihypoesthesia, hemiparesis and mild left facial paresis. CT showed a hypodense region in the right basal ganglia location. The right lateral ventricle seemed mildly compressed due to edematous changes. The magnetic resonance imaging revealed hyperintense signal changes that affected the right lentiform nucleus and the head of the caudate nucleus. The aim of the case is to remind emergency physicians that post-traumatic ischemic stroke is uncommon but may be the cause of disability in pediatric patients, and a systematic physical examination must be performed in all ages even if the patients appear quite well. PMID- 21644102 TI - [Delayed pseudomeningocele as a result of penetrating injury: case report]. AB - Pseudomeningocele is cerebrospinal fluid collection in an extradural area after meningeal tear. The size of the defect in the dura-arachnoid, the pressure of spinal fluid, and the resistance of the soft tissue presumably determine the size of the pseudocyst. The main symptoms are often: headache, neck pain and myelopathic and radicular signs. Pseudomeningocele is diagnosed by myelography, sonography, computed tomography, and magnetic resonance imaging. We present a case of intraoperative identification nerve root entrapment by pseudomeningocele cyst and postoperative recovery of a patient's neurologic deficit. PMID- 21644103 TI - Analgesic effect of Anatolian propolis in mice. AB - OBJECTIVES: The aim of the present study was to assess the analgesic effect of Anatolian propolis, which is added to toothpastes as a prophylactic component for periodontal diseases. METHODS: Water-, ethanol- and acetone-extracted Anatolian propolis were prepared. The analgesic effect of the extracts was assessed using the tail-flick test in mice (n=6 per extract). Comparison among groups was made using one-way ANOVA, followed by post-hoc Scheffe test to determine significant differences among the means of the data groups. P<0.05 was accepted as indicating a significant difference. RESULTS: We found that water-extracted Anatolian propolis caused a significant increase, 1.61-fold (p<0.001 versus control), in the latency time using tail-flick test in mice. However, acetone-extracted and ethanol-extracted propolis led to no significant effect. CONCLUSION: We proved the analgesic effect of water-extracted Anatolian propolis. Thus, propolis used in the composition of toothpastes may be beneficial in terms of its analgesic action in addition to its other favorable effects. PMID- 21644104 TI - Effects of perineural tramadol on nerve conduction of sural nerve. AB - OBJECTIVES: The aim of this study was to investigate whether tramadol had a dose dependent blocking effect on nerve conduction when administered perineurally to the sural nerve of healthy volunteers. METHODS: Twenty-four informed healthy subjects were randomized into four equal groups [Saline (placebo), 0.5% tramadol, 1% tramadol and 1.5% tramadol]. The study was designed to be double-blinded. Sensory nerve action potentials were recorded electroneurographically. Two milliliters of study solution was administered to the sural nerve perineurally at the level of the ankle with the guidance of a nerve stimulator. A sensory block was assumed to have developed when the amplitude of the averaged sensory nerve action potentials diminished below 80% of the baseline value in the subsequent recordings. RESULTS: According to the electroneurographical recordings, none of the volunteers in the saline group had block. However, the block rates with 0.5%, 1% and 1.5% tramadol were 1/6, 4/6 and 6/6, respectively (p<0.05). The maximum decrement in the sensory action potential amplitudes with respect to baseline amplitudes (given as median values) were as follows: 7.8% with saline; 12.5% with 0.5% tramadol; 38.5% with 1% tramadol; and 77.5% with 1.5% tramadol (p<0.05). While the median duration of sensory block with 0.5% tramadol was 5 minutes, it was 15 minutes with 1% tramadol and 35 minutes with 1.5% tramadol. CONCLUSION: Perineurally administered tramadol blocks sensory nerve conduction of the sural nerve in a dose-dependent manner. PMID- 21644105 TI - The effects of morphine and fentanyl alone or in combination added to intrathecal bupivacaine in spinal anesthesia for cesarean section. AB - OBJECTIVES: This randomized double-blind controlled trial examined the effects of fentanyl and morphine, alone and in combination, as adjuncts to spinal anesthesia for elective cesarean section. METHODS: Sixty women undergoing elective cesarean section, with spinal anesthesia using 0.5% hyperbaric bupivacaine, were randomly allocated to receive morphine 0.2 mg, fentanyl 25 ug, or fentanyl 12.5 ug plus morphine 0.1 mg, intrathecally. The start of spinal block, the time to T10 level, the highest sensorial and motor block level, time to regression of sensory block to T10, time to resolution of motor block, surgical characteristics, maternal side effects, Apgar and NACS scores, umbilical blood gas evaluations, and time to first analgesic requirement were recorded. RESULTS: No patient experienced pain during the intraoperative period. The degree and time of sensorial and motor block were similar in both groups, and there was no difference in time to T10 level and time to reversal of motor block. The difference in time to first postoperative analgesic requirement was statistically significant. There was no difference between groups in postoperative side effects. There were no neonatal differences in Apgar and NACS scores or umbilical blood gas evaluations. CONCLUSION: The quality of postoperative analgesia with morphine, when used alone, was found to be superior to that with fentanyl. The combination of opioids offered no advantages over morphine alone. PMID- 21644106 TI - [The relation between pain and pain beliefs and sociodemographic-economic characteristics in an adult population]. AB - OBJECTIVES: In order to control pain, which is a prevalent problem, gathering knowledge regarding pain and pain beliefs is crucial. The aim of this study was to evaluate the correlation between pain and pain beliefs and the sociodemographic and economic characteristics of an adult population. METHODS: This descriptive, cross-sectional study was completed with 131 individuals aged between 18-65 years. A questionnaire evaluating sociodemographic and economic status and pain characteristics and the Pain Beliefs Scale were used for data collection. RESULTS: 78.6% of the research group experienced pain within the last year; of them, 38.8% suffered from chronic pain. According to the results of logistic regression analysis, the risk factors were determined as age between 30 65 years (odds ratio [OR]: 0.215; p=0.008) and graduation from elementary school and lower education level (OR=3.427; p=0.021) for experiencing lifelong frequent pain; being female (OR=3.003; p=0.016) and married (OR=4.550; p=0.005) for experiencing pain within the last year; and age between 30-65 years (OR=3.027; p=0.027) and lower income (OR=4.932; p=0.001) for chronic pain. The organic and psychological pain beliefs scores were similar. Sociodemographic and economic determinants were not significant for the organic subscale (p>0.05), but lower income determined 11% of the psychological subscale (R2=0.115; p<0.05). CONCLUSION: Sociodemographic and economic status were determined to be risk factors for experiencing pain, and cultural factors related to pain beliefs warrant investigation. PMID- 21644107 TI - [Nursing approaches toward postoperative pain in patients: patients' opinions]. AB - OBJECTIVES: We aimed in this defining study to determine patients' opinions about nursing practices with respect to their postoperative pain and their satisfaction with these practices. METHODS: Data were collected by two questionnaires that were prepared by the researchers. The collected data were evaluated with chi square and number, frequency tests. RESULTS: Of the 360 patients participating in this study, 61.4% were women, 83% were over 40 years old and 88% had low educational level. 60.4% had no previous surgery experience. It was determined that 30.6% of patients had undergone heart-lung (cardiovascular system, CVS) and 25.5% gastrointestinal system (GIS) surgery. It was found that 96.4% of patients experienced pain while coughing and 81.9% while getting out of bed. 96.4% of patients reported difficulty in coughing, 78.3% difficulty in mobilization and 46.7% difficulty in breathing because of postoperative pain. It was determined that patients who underwent CVS, TAH+BSO and GIS operations experienced more pain while coughing and trying to get out of bed, while patients who underwent discectomy experienced more pain while walking. There was a significant difference between types of operation and pain (p<0.05) and between activities in which patients experienced difficulty and types of operation (p<0.05). CONCLUSION: The patients stated that nurses did not use a pain scale, provide information about how pain could be relieved, demonstrate what they needed to do during the activities that caused pain, assist the patients to reposition to relieve the pain, or use any non-pharmacological interventions. Nevertheless, all of the patients were satisfied with the analgesics that were administered by nurses for pain relief. PMID- 21644108 TI - Possible heart failure associated with pregabalin use: case report. AB - Pregabalin and gabapentin are widely used analgesic, anticonvulsant and anxiolytic agents as they are relatively reliable and easily tolerated. However, they may cause some side effects such as dizziness, somnolence, dose-dependent peripheral edema, and weight gain, which may cause patients to abandon their use. Furthermore, there are a few reports in the literature addressing elderly patients with serious chronic disease and cardiac history, who develop heart failure during pregabalin application. In this report, we present a patient with no cardiac history treated with 300 mg/kg pregabalin due to neuropathic pain, who developed peripheral and then central edema, which were determined after advanced investigations. After stopping pregabalin, the situation regressed. Then, peripheral edema developed associated with the recommended dose of gabapentin, which was used in place of pregabalin. Despite the lack of any published evidence, the New York Heart Association issued a warning about using caution when prescribing pregabalin to type III-IV heart failure patients. Though the effect mechanisms of pregabalin and gabapentin are not well known, the calcium channel relationship may lead to these side effects. In summary, we believe that pregabalin and gabapentin, which is mostly used nowadays, should be administered with care not only in patients with advanced cardiac pathology but also in all patients, due to the potential side effects. PMID- 21644109 TI - [Modified van-Lint technique for the management of postherpetic neuralgia: case report]. AB - Acute herpes zoster (AHZ) causes postherpetic neuralgia (PHN) in 48-75% of patients. Nerve blocks performed in the acute phase of HZ may treat the pain and prevent PHN development. Here, we present pain relief with modified van-Lint block in two cases with AHZ involving vesicles on the traces of the supraorbital and supratrochlear nerves. Two female patients, 72 and 66 years old, with AHZ involving vesicles on the traces of the supraorbital and supratrochlear nerves starting from the right periocular region to the scalp presented with symptoms such as hypoesthesia, dizziness, burning, throbbing, and severe pain. Their initial visual analogue scale (VAS) scores for pain were 9 and 10, respectively. Supraorbital and supratrochlear nerve blockade with modified van-Lint technique was planned, as the classical nerve block sites were covered with active vesicles. Following the nerve blocks, VAS scores of both patients decreased to 1 immediately. Vesicles were faded and scabbed, symptoms such as hypoesthesia, burning and throbbing had recovered, dizziness was relieved, and VAS scores were 4 and 5, respectively, after one week. VAS scores were 1 and 2, respectively, after the second injection, and all symptoms were resolved, and no additional analgesic was required during a three-month follow-up. Modified van-Lint block with 5 mL 1% lidocaine may provide successful pain relief in AHZ involving vesicles on the traces of the supraorbital and supratrochlear nerves. PMID- 21644110 TI - [Pasha-Cath in the treatment of neuropathic pain: case report]. AB - In neuropathic pain, the nerve fibers themselves may be damaged, dysfunctional or injured. A 49-year-old female patient experienced radiating right anterior arm pain following an operation on the same arm. Visual analogue scale (VAS) score was 10. Tramadol, gabapentin and amitriptyline were started as medical treatment. In view of the persisting complaints, epidural pulsed radiofrequency with Pasha Cath was scheduled. After 2 years, VAS remained at 1-2. It was concluded that epidural pulsed radiofrequency with Pasha-Cath is an alternative and effective choice of treatment in neuropathic pain when the medical treatment alone is not sufficient. PMID- 21644111 TI - The role of holistic care culture in mitigating burnout and enhancing engagement: a study among elderly service workers in Hong Kong. AB - OBJECTIVES: Elderly service work is a labor intensive and emotion demanding occupation. Workers in this field are prone to burnout, a form of emotional exhaustion at work. While their job well-being is associated with a number of job demands, little research has been done in exploring job resources to promote their well-being. Holistic care culture (HCC) was proposed as an organizational culture of holistic caring. This study explored the role of HCC in predicting job well-being and moderating the impact of perceived stress on job well-being. METHOD: A large-scale questionnaire survey was conducted among 992 elderly service workers of a major social service organization in Hong Kong, with a 93% response rate. Participants completed a standardized self-report questionnaire. RESULTS: Structural equation modeling found HCC negatively predicted burnout (beta = -0.32, p < 0.01) and positively predicted engagement (beta = 0.36, p < 0.01). HCC showed significant moderating on the relationship between perceived stress and job well-being in expected directions. The model explained 53.7% and 35.8% of variance in burnout and engagement. CONCLUSION: Findings from this study provides supportive evidence for HCC as a moderator to ameliorate burnout and facilitate engagement among elderly service workers. Further studies of rigorous design on HCC are recommended. Implication of this study for organizational practice was discussed. PMID- 21644112 TI - Is academic buoyancy anything more than adaptive coping? AB - Academic buoyancy refers to a positive, constructive, and adaptive response to the types of challenges and setbacks experienced in a typical and everyday academic setting. In this project we examined whether academic buoyancy explained any additional variance in test anxiety over and above that explained by coping. Two hundred and ninety-eight students in their final two years of compulsory schooling completed self-report measures of academic buoyancy, coping, and test anxiety. Results suggested that buoyancy was inversely related to test anxiety and unrelated to coping. With the exception of test-irrelevant thoughts, test anxiety was positively related to avoidance coping and social support. Test irrelevant thoughts were inversely related to task focus, unrelated to social support, and positively related to avoidance. A hierarchical regression analysis showed that academic buoyancy explained a significant additional proportion of variance in test anxiety when the variance for coping had already been accounted for. These findings suggest that academic buoyancy can be considered as a distinct construct from that of adaptive coping. PMID- 21644113 TI - Measurement of anxious traits: a contemporary review and synthesis. AB - Trait anxiety is a characteristic predisposition to appraise stimuli as threatening and respond with anxiety. Trait anxiety is proposed to serve as a vulnerability factor for greater frequency and intensity of anxiety experiences as well as the development of anxious pathology. Cognitive, behavioral, and physiological components of trait anxiety have been described. Common self-report measures of trait anxiety are reviewed with an emphasis on: components assessed, factor structure, internal consistency, reliability, and construct validity. Measures were selected if instructions ask individuals to endorse items based on their characteristic, rather than a time sensitive, response. Selection criteria resulted in a focus on the following measures: the State-Trait Anxiety Inventory (STAI), Beck Anxiety Inventory-Trait (BAIT), Cognitive Somatic Anxiety Questionnaire (CSAQ), Endler Multidimensional Anxiety Scales-Trait (EMAS-T), Four Systems Anxiety Questionnaire (FSAQ), State-Trait Inventory for Cognitive and Somatic Anxiety (STICSA), and the Three Systems Anxiety Questionnaire (TSAQ). While the STAI has the advantage of normative data and frequent use in prior research, newer measures, such as the BAIT and the STICSA, demonstrate greater discriminant validity. The strengths and weaknesses of each of the reviewed measures are highlighted. Recommendations for measure selection and future research are provided. PMID- 21644114 TI - The effect of feed salinity on the biofouling dynamics of seawater desalination. AB - A persistent cell labeling dye and a novel microbial counting method were used to explore the effects of salinity on a microbial population in a reverse osmosis (RO) desalination system, and these clearly distinguished microbial cell multiplication from cell adherence. The results indicated that microbial multiplication is more active at the front of a seawater RO pressure vessel, while adhesion dominates the back of the vessel. A severe reduction in RO permeate flux and total dissolved solid (TDS) rejection were detected at low salinity, attributed to marked cell multiplication and release of extracellular polymeric substances, whilst a relatively stable flux was observed at medium and high salinity. The results from PCR-DGGE revealed the variation in microbial species distribution on the membrane with salinity. The results imply the critical role of membrane modification in biofouling mitigation in the desalination process. PMID- 21644115 TI - Modelling transport of layered double hydroxide nanoparticles in axons and dendrites of cortical neurons. AB - This paper develops a model of nanoparticle transport in neurons. It is assumed that nanoparticles are transported inside endocytic vesicles by a combined effect of dynein-driven transport and diffusion. It is further assumed that in axons nanoparticles are internalised only at axon terminals, whereas in dendrites nanoparticles can enter through the entire plasma membrane. This causes differences in transport of nanoparticles in axons and dendrites; these differences are investigated in this paper. Another difference is microtubule (MT) orientation in axons and dendrites; in axons, all MTs have their plus-ends oriented towards the axon terminal; in a proximal region of a dendrite, MTs have mixed orientation, whereas in a distal dendritic region the MT orientation is similar to that in an axon. It is shown that if molecular-motor-driven transport were powered by dynein alone, such MT orientation in a dendrite would result in a region of nanoparticle accumulation located at the border between the proximal and distal dendritic regions. PMID- 21644116 TI - Gay men and ambivalence about 'gay community': from gay community attachment to personal communities. AB - The concept of 'gay community', and gay men's attachment to and involvement in gay community activities, has held both a symbolic and practical role in understanding and guiding responses to HIV in developed world contexts. In the West, the HIV epidemic has disproportionately affected gay men. Being involved in and connected to gay community activities (what, in Australia, is described as 'gay community attachment') predicted the adoption of safe sex practices. However, the meaning of gay community is changing. This presents a challenge to those working in HIV prevention. With reference to previous research, the meaning of gay community is analysed in qualitative interviews conducted with Australian gay men. The interview data indicate that gay men are often ambivalent about gay communities, suggesting a need for subtlety in the ways we think about and address gay men in HIV education and health promotion. The concept of 'personal communities' may better reflect the ways in which gay men engage with each other and their social networks. Recognising and responding to the changing nature of gay life will ensure that the flexibility and pragmatism of HIV programmes aimed at gay men are maintained. PMID- 21644117 TI - Investigating reasons for ethnic inequalities in breast cancer survival in New Zealand. AB - OBJECTIVE: This study investigated the role that demographic and tumour factors play in explaining ethnic inequalities in breast cancer survival. DESIGN: Breast cancer cases notified to the New Zealand Cancer Registry (NZCR) from April 2005 to April 2007 were followed up to April 2009. Maori, Pacific and non-Maori/non Pacific women were categorised according to ethnicity on the NZCR. Deprivation was analysed as quintiles of the New Zealand area-based index of socio-economic position. Relative survival rates were estimated using ethnic-specific life tables. Missing values were imputed and excess mortality modelling was used to estimate the contribution of demographic and tumour factors to ethnic inequalities in survival. RESULTS: There were 2968 breast cancer cases (76.5% non Maori/non-Pacific, 17% Maori, and 6.5% Pacific) included and 433 recorded deaths. Relative survival rates at 4 years were 91.5% (95% confidence interval (CI) 89.7 to 92.9) for non-Maori/non-Pacific, 86.2% (CI 80.3 to 90.4) for Maori, and 79.6% (CI 68.2 to 87.2) for Pacific women. Using non-Maori/non-Pacific as the reference group, the age-adjusted hazard ratio (HR) dropped for Maori from 1.76 (CI 1.22 to 2.48) to 1.43 (CI 0.97 to 2.10) when further adjusted by deprivation. For Pacific the HR dropped from 2.49 (CI 1.57 to 3.94) to 1.94 (CI 1.20 to 3.13). Inequalities persisted after adjustment for subtype variables (ER/PR/HER2), but adjusting for access to care variables (extent/size) eliminated the ethnic inequalities in excess mortality. CONCLUSION: Ethnic disparities in breast cancer survival in New Zealand can be attributed to deprivation and differential access to health care rather than differences in breast cancer subtypes. PMID- 21644118 TI - Modified dispersive liquid-liquid microextraction followed by high-performance liquid chromatography for the determination of clenbuterol in swine urine. AB - A modified dispersive liquid-liquid microextraction (DLLME) technique combined with an HPLC-UV procedure was developed for the extraction and determination of clenbuterol in swine urine. The modification involved the selection of methyl tert-butyl ether (MTBE) as the dispersive solvent, which had a low solubility in aqueous samples, playing the part of dispersion with the help of violent shaking. MTBE improved the partition of clenbuterol into the extractant, and helped the formation of phase separation. Various factors affecting the extraction efficiency including selection of the organic extractant and the dispersive solvent, the volume of extractant and dispersive solvent, salt concentration, NaOH concentration and centrifugation time were evaluated and optimised. Under the optimal conditions, precision, linearity (correlation coefficient, r (2 )= 0.996 over the concentration range of 10-1000 ng ml(-1)), detection limit (2.4 ng ml(-1)) and enrichment factor of 55 were obtained. The modification to the DLLME made it suitable for analytes with pronounced solubility, especially when the compounds are highly polar and thus more difficult to extract effectively by DLLME. The procedure was suitable for the fast screening of clenbuterol residue in swine urine. PMID- 21644119 TI - Sphinganine and sphingosine levels and ratio in urine and blood samples from a Catalonian population, Spain. AB - Fumonisins occur mainly in maize and they produce alterations on sphingolipid metabolism, unbalancing the sphinganine (Sa)/sphingosine (So) ratio. This alteration has been proposed as a biomarker of fumonisin exposure. The objective of this study was to establish the urinary and plasmatic levels of Sa, So as well as the ratio Sa/So from a sample of the Catalonian (Spain) population exposed to fumonisins at low levels. Firstly, plasma and urinary Sa and So levels and the ratio Sa/So were compared between two population groups, and later urinary Sa and So levels from corn food consumers and a control group were monitored for 2 weeks under controlled intake of corn foods. Sa and So levels were determined in urine and blood samples using validated methods using HPLC with fluorescence detection. Significant differences were not found in urine samples when Sa/So ratios were compared from corn food consumers and non-consumers, while significant differences were found in urine and plasma samples, but evidence of the mechanism of action of fumonisins was not apparent. Through a time-course study we have narrowed down the day in which the maximum alteration of Sa/So ratio should be expected in humans. This paper reports some useful information to improve the design of studies to validate the ratio Sa/So as a possible biomarker of fumonisin exposure. PMID- 21644120 TI - Challenging household dynamics: impact of orphanhood, parental absence, and children's living arrangements on education in South Africa. AB - Public health and social-historical changes have had multiple effects on South African children and families. This study examines the association between challenging family dynamics, such as child orphan status, and educational delay, as defined by being below proper grade-for-age. Analysing the 2003/2004 South Africa Demographic and Health Surveys (DHS) dataset, we estimate orphan prevalence, maternal and paternal household presence, other household characteristics, and schooling variables in a nationally representative household sample. Among 5592 children ages 8-14 in South Africa, 21% had experienced parental death, 33% did not have their mother present in the household, and 63% of the children were not living with their father. Twenty per cent were behind proper grade-for-age. A bivariate analysis shows that orphaned children experienced 35% greater odds of being behind in school (p<0.001). After adjusting for parental presence, household characteristics, and socio-demographic factors, orphan status does not remain significantly associated with being behind in school. However, maternal presence, relationship to the household head, number of children in a household, and socio-demographic characteristics each independently affect a child's likelihood of educational delay. Findings offer a more nuanced understanding of household dynamics that may protect against or exacerbate educational delays among vulnerable youth. PMID- 21644121 TI - The sociocognitive and dissociation theories of hypnosis: toward a rapprochement. AB - In this introductory article to a special issue on the sociocognitive perspective of hypnosis, the authors contrast two influential hypnosis theories-the sociocognitive and dissociation perspectives-and argue that recent developments in sociocognitive theory (i.e., response set theory) and in the broader field of cognitive psychology pertaining to nonconscious information processing and goal directed action make possible a rapprochement between theoretical accounts that have vied for attention and empirical support. PMID- 21644122 TI - The impact of hypnotic suggestibility in clinical care settings. AB - Hypnotic suggestibility has been described as a powerful predictor of outcomes associated with hypnotic interventions. However, there have been no systematic approaches to quantifying this effect across the literature. This meta-analysis evaluates the magnitude of the effect of hypnotic suggestibility on hypnotic outcomes in clinical settings. PsycINFO and PubMed were searched from their inception through July 2009. Thirty-four effects from 10 studies and 283 participants are reported. Results revealed a statistically significant overall effect size in the small to medium range (r = .24; 95% Confidence Interval = 0.28 to 0.75), indicating that greater hypnotic suggestibility led to greater effects of hypnosis interventions. Hypnotic suggestibility accounted for 6% of the variance in outcomes. Smaller sample size studies, use of the SHCS, and pediatric samples tended to result in larger effect sizes. The authors question the usefulness of assessing hypnotic suggestibility in clinical contexts. PMID- 21644123 TI - Are high hypnotizables especially vulnerable to false memory effects? A sociocognitive perspective. AB - This article examines issues raised by a recent UK legal case in which the defense argued that the accusations made by the highly hypnotizable plaintiff were likely based on false memories. The authors argue that the evidence related to hypnotizability and false memory production is inconsistent but may be illuminated by a sociocognitive perspective. They present 2 preliminary studies that indicate that when the instructions imply that accurate reporting is a feature of hypnosis, higher hypnotizables may actually be more resistant than low or medium hypnotizables to false memories arising from misleading information given during hypnosis. They conclude that, when memory accuracy is emphasized rather than productivity, there is little evidence to link high hypnotizability with a propensity to produce false memories. PMID- 21644124 TI - Responding to hypnotic and nonhypnotic suggestions: performance standards, imaginative suggestibility, and response expectancies. AB - This study examined the relative impact of hypnotic inductions and several other variables on hypnotic and nonhypnotic responsiveness to imaginative suggestions. The authors examined how imaginative suggestibility, response expectancies, motivation to respond to suggestions, and hypnotist-induced performance standards affected participants' responses to both hypnotic and nonhypnotic suggestions and their suggestion-related experiences. Suggestions were administered to 5 groups of participants using a test-retest design: (a) stringent performance standards; (b) lenient performance standards; (c) hypnosis test-retest; (d) no-hypnosis test retest; and (e) no-hypnosis/hypnosis control. The authors found no support for the influence of a hypnotic induction or performance standards on responding to suggestions but found considerable support for the role of imaginative suggestibility and response expectancies in predicting responses to both hypnotic and nonhypnotic suggestions. PMID- 21644125 TI - The altered state issue: dead or alive? AB - Theoretical positions on the altered-state issue are viewed on a continuum rather than a dichotomy. While differences between some pairs of positions have little or no substantive interest, others are important to understanding the nature of hypnotic phenomena. Recent brain imaging data from the University of Hull are reviewed with respect to their implications concerning the existence and functional significance of the hypothesized hypnotic state. PMID- 21644126 TI - Does neuroimaging of suggestion elucidate hypnotic trance? AB - Contemporary studies in the cognitive neuroscience of attention and suggestion shed new light on the underlying neural mechanisms that operationalize these effects. Without adhering to important caveats inherent to imaging of the living human brain, however, findings from brain imaging studies may enthrall more than explain. Scholars, practitioners, professionals, and consumers must realize that the influence words exert on focal brain activity is measurable but that these measurements are often difficult to interpret. While recent brain imaging research increasingly incorporates variations of suggestion and hypnosis, correlating overarching hypnotic experiences with specific brain substrates remains tenuous. This article elucidates the mounting role of cognitive neuroscience, including the relative merits and intrinsic limitations of neuroimaging, in better contextualizing trance-like concepts. PMID- 21644127 TI - Repeated holdout cross-validation of model to estimate risk of Lyme disease by landscape characteristics. AB - We previously modeled Lyme disease (LD) risk at the landscape scale; here we evaluate the model's overall goodness-of-fit using holdout validation. Landscapes were characterized within road-bounded analysis units (AU). Observed LD cases (obsLD) were ascertained per AU. Data were randomly subset 2,000 times. Of 514 AU, 411 (80%) were selected as a training dataset to develop parameter estimates used to predict observations in the remaining 103 (20%) AU, the validation subset. Predicted values were subtracted from obsLD to quantify accuracy across iterations. We calculated the percentage difference of over- and under-estimation to assess bias. Predictive ability was strong and similar across iterations and datasets; the exact number of obsLD cases per AU were predicted almost 60% of the time. However, the three highest obsLD AU were under-predicted. Our model appears to be accurate and relatively unbiased, however is conservative at high disease incidence. PMID- 21644128 TI - Ambient air pollution is associated with the increased incidence of breast cancer in US. AB - Women in the United States have among the highest incidence rates of breast cancer. The reasons behind this are not fully understood. In this study we analyzed US ecological data to examine the effect of ambient air pollution on breast cancer incidence. Time trends and regional variations in breast cancer incidence were assessed in relation to emissions of air pollutants. A statistically significant increase in the incidence of female breast cancer in US was observed during 1986-2002, which could occur following the increased emissions of air pollutants as a result of industrial development and automobile use. Emissions of nitrogen oxides, carbon monoxide, sulfur dioxide, and volatile organic compounds were shown to be positively associated with breast cancer incidence with r = 0.89, 0.82, 0.71, and 0.68, respectively (p < 0.001). A higher incidence rate of breast cancer was found in high emission regions and metropolitan areas. This study suggests a possible association between air pollution and female breast cancer in US. PMID- 21644129 TI - Short-term impact of particulate matter (PM(2.5)) on respiratory mortality in Madrid. AB - OBJECTIVES: This paper sought to quantify the particulate matter (PM(2.5)) pollutant's impact on short-term daily respiratory-cause mortality in the city of Madrid. METHODS: As our dependent variable, we took daily mortality registered in Madrid from 1 January 2003 to 31 December 2005, attributed to all diseases of the respiratory system as classified under heads J00-J99 of the ICD 10 and broken down as follows: J12-J18, pneumonia; J40-J44, chronic diseases of the respiratory system except asthma; J45-J46, asthma; and J96, respiratory failure. RESULTS: The relative risk (RR) for daily overall respiratory mortality was RR 1.0281 (1.0043 1.0520), with a proportional attributable risk (PAR) of 2.74%. This effect occurred in lag 1; respiratory failure, RR 1.0816 (1.0119-1.1512) and PAR 7.54% at lag 5; and pneumonia, RR 1.0438 (1.0001-1.0875) and PAR 4.19% at lag 6. CONCLUSIONS: Our results reflect the association that exists between PM(2.5) concentrations and daily respiratory-cause mortality. PMID- 21644130 TI - Lost papers. PMID- 21644131 TI - Reprint of a lost paper FAO/IAEA - interregional training course on the use of (15)N in soil science and plant nutrition. Training manual. PMID- 21644132 TI - Galena oxidation investigations on oxygen and sulphur isotopes. AB - Batch experiments with the lead sulphide ore mineral galena were carried out in order to get information about the oxidation mechanisms and to contribute to the understanding of field data, especially those obtained from the mining-affected sites. Results indicate that oxygen and sulphur isotopes of dissolved sulphate may be useful tools for the investigation of galena oxidation mechanisms. However, some methodological modifications are necessary to obtain sufficient sulphate yields, which are a prerequisite for the reduction of the analytical uncertainty. Surface and hydrochemical investigations indicated that galena was dissolved non-oxidatively during the experiment at initial pH 2, whereas the oxidative dissolution of galena dominated in experiments at initial pH 6 and 8. No isotope results could be obtained from the experiment at initial pH 2 due to the low sulphate production. The oxidative dissolution of galena resulted in somewhat higher sulphate yields. Thus, the oxygen isotope composition of sulphate produced from galena oxidation could be determined for the first time. Oxygen in sulphate derived largely from water molecules; but minor amounts of molecular oxygen could be also observed. Both molecular oxygen and ferric iron may act as oxidants of galena. A variety of intermediate reactions allow for a variable oxygen isotope composition of sulphate. Sulphur isotopes of sulphate showed an enrichment of (32)S in sulphate (relative to galena), which increases with increasing pH. Sulphur isotope enrichment processes may be associated with the formation of anglesite. PMID- 21644133 TI - Relationship between isotope composition of sulphur in sulphate dissolved in river water and sulphur extracted from fish scales. AB - More than 100 trouts captured in more than 20 rivers from Poland have been analysed. Only fish caught by trout fishermen were used for this study. Isotope compositions of sulphur (delta(34)S) and carbon (delta(13)C) of fish scales were examined together with delta(34)S from sulphate dissolved in river, as well as patterns of fish diet. We predominantly examined adult fish, at least 4 years old. A scatter of isotope values occurred among the samples obtained from fish caught in different rivers and also for fish caught in the same river. The scatter of the delta(34)S values was much larger than that of delta(13)C values. We noticed the relationship between delta(34)S of scales and delta(34)S of riverine sulphate. There is also a significant difference between isotope compositions obtained for different fish species, which can be attributed to their different diet. PMID- 21644134 TI - Nitrogen isotopic signature of vegetables from the Slovenian market and its suitability as an indicator of organic production. AB - The aim of this study was to obtain preliminary data on delta(15)N values in organically and conventionally grown vegetables available on the Slovenian market to determine whether there are any characteristic differences in delta(15)N values, which could be used as a marker to differentiate between organically and conventionally grown vegetables, namely vegetables grown with synthetic or organic nitrogen fertilisers. Therefore, 14 different varieties of organically and conventionally grown vegetables were analysed. The results obtained show clear differences in mean delta(15)N values between organically and conventionally grown vegetables (up to 6.3?permil). However, due to overlapping results, it was not possible to differentiate between organically and conventionally grown counterparts in eight vegetable varieties, despite the differences in mean delta(15)N values. The results obtained indicate that the method has several limitations. Hence, delta(15)N could be a reliable indicator for organic production, but nevertheless, the conclusion has to be validated by background information. PMID- 21644135 TI - Soil respiration rates and delta13C(CO2) in natural beech forest (Fagus sylvatica L.) in relation to stand structure. AB - Soil respiration rates were studied as a function of soil type, texture and light intensity at five selected natural beech forest stands with contrasting geology: stands on silicate bedrock at Kladje and Bricka in Pohorje, a stand on quartz sandstone at Vrhovo and two stands on a carbonate bedrock in the Karstic-Dinaric area in Kocevski Rog, Snezna jama and Rajhenav, Slovenia, during the growing season in 2005-2006. Soil respiration exhibited pronounced seasonal and spatial variations in the studied forest ecosystem plots. The CO(2) flux rates ranged from minimum averages of 2.3 MUmol CO(2) m(-2) s(-1) (winter) to maximum averages of about 7 MUmol CO(2) m(-2) s(-1) (summer) at all the investigated locations. An empirical model describing the relationship between soil respiration and soil temperature predicted seasonal variations in soil respiration reasonably well during 2006. Nevertheless, there were also some indications that soil moisture in relation to soil texture could influence the soil CO(2) efflux rates in both sampling seasons. It was shown that spatial variability of mean soil respiration at the investigated sites was high and strongly related to root biomass. Based on the [image omitted] data, it was shown that new photoassimilates could account for a major part of the total soil respiration under canopy conditions in forest ecosystems where no carbonate rocks are present, indicating that microbial respiration could not always dominate bulk soil CO(2) fluxes. At Snezna jama and Rajhenav, the abiotic CO(2) derived from carbonate dissolution had a pronounced influence on CO(2) efflux accounting, on average, to ~17%. Further spatial heterogeneity of soil respiration was clearly affected by management practice. Higher respiration rates as well as higher variability in respiration rates were observed in the virgin forest (Rajhenav) than in the management forest (Snezna jama) and could be related to a higher amount of detritus and consequently to less pronounced influence of inorganic pool to CO(2) efflux, lower mixing with atmospheric CO(2) and higher sensitivity to environmental changes. Major differences in soil carbon dynamics among these five forest ecosystems can be explained by the influence of bedrock geology (particularly, the presence or absence of carbonate minerals) and soil texture (affecting gas exchange with overlying air and soil moisture). PMID- 21644136 TI - Measuring technique for thermal ionisation mass spectrometry of human tracer kinetic study with stable cerium isotopes. AB - Thermal ionisation mass spectrometry (TIMS) method has been developed for the simultaneous detection of different cerium isotopes in biological samples (i.e., blood and urine) at very low concentrations. The work has been done in the frame of a biokinetic study, where different stable cerium isotopes have been administered orally and intravenously as tracers to the human body. In order to develop an appropriate detection method for the tracers in the biological samples, an optimum sample preparation technique has been set and adapted to the specific requirements of the analysis technique used, i.e., TIMS. For sample evaporation and ionisation, the double tantalum filament technique showed the best results. The ions produced were simultaneously collected on a secondary electron multiplier so that the isotopic ratios of the cerium isotopes in the biological samples could be measured. The technique has been optimised for the determination of cerium down to 1 ng loaded on the evaporation filament corresponding to cerium concentrations of down to 1 ng ml(-1) in the blood or urine samples. It has been shown that the technique is reliable in application and enables studies on cerium metabolism and biokinetics in humans without employing radioactive tracers. PMID- 21644138 TI - The effect of semantic orientation at encoding on free-recall performance in amnestic mild cognitive impairment and probable Alzheimer's disease. AB - The present study manipulated the nature of orientation provided at encoding in an intentional word memory task. Performance on the memory task was then compared between 23 elderly persons with amnestic mild cognitive impairment (aMCI), 13 patients with probable Alzheimer's disease (AD), and 23 healthy elderly persons. When tested following shallow (reading orientation) encoding, free-recall performance was impaired in AD compared to aMCI and healthy older adults. When tested following deep (categorical semantic orientation) encoding, both AD and aMCI groups were impaired relative to healthy older adults. The latter result was related to larger memory improvement due to semantic orientation in healthy controls than in aMCI and AD participants. Overall, these findings indicate that the encoding put up by aMCI and healthy elderly persons is comparably efficient in situations where shallow supportive cues are provided at encoding, but that healthy controls benefit more than aMCI and AD in situations where supportive cues are strong. PMID- 21644139 TI - Cognitive profiles in persons with chronic schizophrenia. AB - Cognitive heterogeneity has been a key barrier to clarifying the neuropathologic underpinnings of schizophrenia. We used an idiographic method for cluster analysis of neuropsychological data from 144 middle-aged and older people with schizophrenia to characterize and group the patterns of relative (within-person) profiles of cognitive strength and weakness. Results indicated a 5-cluster solution as most appropriate, with relatively even distribution across the 5 clusters in terms of the proportion of patients in each cluster. Cognitive subtyping may be useful in imaging and genetic research on schizophrenia, as well as having practical utility in treatment planning and cognitive rehabilitation. PMID- 21644140 TI - Modeling longitudinal change in motor and cognitive processing speed in presymptomatic Huntington's disease. AB - Persons who have the genetic mutation responsible for Huntington's disease (HD) have been shown to exhibit lower cognitive test scores years prior to manifest HD. Most studies have examined cognitive performance in presymptomatic persons by using estimated times to manifest HD based on published algorithms. We followed 19 gene-positive persons who were presymptomatic using an objective criterion (i.e., Quantified Neurological Exam score <= 10) at baseline for up to 21 years (median = 5 years) with periannual neuropsychological assessments until a diagnosis of manifest HD. Results showed that our tests of information- and psychomotor-processing speed that place minimal demands on cognition, worsening performance became evident 5-10 years prior to the development of manifest HD. In conclusion, cognitive decline precedes motor signs in HD and may be an important target in clinical trials and early intervention. Cognitive test scores may also improve the ability to predict disease onset among gene mutation carriers and help families to better plan for potential personal and economic hardship. PMID- 21644141 TI - Assessing cognitive impairment following stroke. AB - The assessment of cognitive function is often neglected following stroke, with no consensus on the optimal method to assess poststroke cognition. We evaluated the ability of a brief protocol to detect cognitive impairment in community-dwelling people with chronic stroke compared to healthy controls and its ability to detect changes in cognition in stroke participants undergoing an exercise intervention. Four tests of cognition were able to detect differences between the groups in the domains of executive function, memory, and information-processing speed. Stroke survivors undergoing exercise over a 5-month period showed significantly improved memory and speed of information processing. Results suggest that exercise may have the potential to improve cognition in long-term stroke survivors and that these tests are sensitive measures of poststroke cognition. PMID- 21644143 TI - Special issue: Constructed wetland technology for water pollution control. Foreword. PMID- 21644144 TI - Modeling E.coli fate and transport in treatment wetlands using the water quality analysis and simulation program. AB - Treatment wetlands can be a viable wastewater treatment option, especially in rural and remote regions where centralized wastewater treatment is not feasible. Bacteria fate and transport modeling within wetlands requires further development if they are to become a more reliable and predictable form of wastewater treatment. The goal of this paper was to calibrate and test an unsteady state numerical model for the simulation of E. coli fate and transport within full scale surface flow (SF) wetlands treating domestic wastewater. The Water Quality Analysis and Simulation Program (WASP) was used to develop the model. Accurately predicting E. coli effluent concentrations using WASP was difficult due to the dynamic nature of the wetland environment including hydraulics, seasonal variability, and wetland maturity. WASP was successful in predicting average E. coli effluent concentrations but did not accurately forecast maximum and minimum values. The model produced better fits with observed E. coli effluent concentrations during the summer months, when observed effluent concentrations were less variable. Hydraulic tracer studies and model results suggest that preferential flow pathways may be affecting E. coli removal due to reduced retention times. Flow channelling or short circuiting may have been caused by high flow conditions and/or dense cattail growth. A more detailed understanding of treatment wetland hydraulics is required before we can accurately predict treatment performance. PMID- 21644145 TI - Performance and modeling of a vertical flow constructed wetland-maturation pond system. AB - A 32-month monitoring program is presented in a vertical flow constructed wetland facility, located in North Greece. The monitoring campaigns were organized every 15 days. Water quality samples were collected at the inlet, at four intermediate points (i.e., at the end of each treatment stage) and at the outlet of the system. Temperature, electrical conductivity, pH and dissolved oxygen (DO) were measured in-situ with the use of appropriate instruments at the same points of water sample collection. Water samples were analyzed for biochemical oxygen demand (BOD), chemical oxygen demand (COD), total Kjeldahl nitrogen (TKN), ammonia, nitrate, nitrite, total phosphorus (TP), ortho-phosphate (OP), total suspended solids (TSS) and total coliforms (TC). Mean removal efficiencies for the monitoring period were: 90.8% for BOD, 89.0% for COD, 83.9% for TKN, 83.8% for ammonia, 38.8% for TP, 17.4% for OP, 90.4% for TSS and 99.9% for TC, indicating, for most pollutant, excellent performance of this constructed wetland (CW) system under Mediterranean climate conditions. Results showed that organic matter, TSS, TKN, ammonia, TP, OP and TC removal is not significantly affected by temperature. The collected removal data were used to produce appropriate parameter values for first-order k-C* models and develop simple models based on stepwise multiple linear regression (SMLR) analysis, in an effort to predict CW performance. These models were verified using data from another facility located in the same region. The results showed that the predictions correlate well with measured values, leading to the conclusion that the first-order models and the developed SMLR models are useful tools in the design of vertical flow (VF) CWs. PMID- 21644146 TI - Treatment of reactive azo dye from textile wastewater by burhead (Echinodorus cordifolius L.) in constructed wetland: Effect of molecular size. AB - The potential of burhead (Echinodorus cordifolius L.) for the treatment of textile wastewater has been investigated. Reactive red 2; RR2 [MW=615], reactive red 120; RR120 [MW=1469] and reactive red 141; RR141 [MW=1775] were studied in order to determine the effect of molecular size on the efficiency of dye removal by plants in batch systems of constructed wetlands under soil and soil-free conditions. Dye concentrations, total dissolve solids (TDS), conductivity and pH in the effluents, and the relative growth rates (RGR) of plants were measured. The highest efficiency of dye removal during 7 days under soil-free conditions was RR2 (33.09 MUmol(RR2)kg(-1)(FW)), followed by RR120 (13.35 MUmol(RR120)kg( 1)(FW)) and RR141 (10.57 MUmol(RR141)kg(-1)(FW)), respectively. This suggests that the structure and size of dye molecule strongly affects the efficiency of dye removal by plant. The results from a synthetic wetland experiment found that dye removal was 96 % at 4 days and 6 days under soil and soil-free conditions, respectively. Furthermore, plants were able to decrease TDS (42 %), conductivity (50 %) and pH (from 9.5 to 7.4) within 2 days in the synthetic reactive red(141) dye wastewater (SRRW141) under soil-free conditions, thus demonstrating the potential of burhead for textile wastewater treatment. PMID- 21644147 TI - Anammox bacteria community and nitrogen removal in a strip-like wetland in the riparian zone. AB - A strip-like wetland was constructed in the riparian zone for investigation of ammonium nitrogen (NH(3)-N) removal in the Peach River. An inner zeolite layer was set in the wetland to adsorb NH(3)-N and further to remove total nitrogen (TN). An oxygen-deficient condition with dissolved oxygen of 0.87-1.60 mg L(-1) was observed in the zeolite layer, which benefits anaerobic ammonium oxidation (anammox) bacteria survival. The community structure of anammox bacteria was analyzed in the zeolite layer. The analysis shows that the anammox bacterial sequences are grouped into three known distinct clusters: Candidatus Brocadia fulgida, Candidatus Brocadia anammoxidans and Candidatus Jettenia asiatica. The intensified test driven by artificial pumping shows that average removal rates of NH(3)-N and TN are 41.6 mg m(-3)d(-1) and 63.2 mg m(-3)d(-1), respectively. The normal test driven by natural hydrodynamics also verifies that NH(3)-N removal mainly happens in the zeolite layer. Microbial mechanism of TN removal in the wetland involves both the autotrophic and heterotrophic process. These results suggest that the strip-like RW can be a cost-effective approach for NH(3)-N removal and can potentially be extended to similar rivers as no extra energy is required to maintain the wetland operation. PMID- 21644148 TI - On-site wastewater treatment using subsurface flow constructed wetlands in Ireland. AB - The results from an Irish EPA-funded project on the effectiveness of using constructed wetlands for treating wastewater from single households is presented, which has contributed to the design guidelines included in the new EPA Code of Practice. Three subsurface flow gravel-filled wetlands were constructed on separate sites--one to provide secondary treatment and the other two to provide tertiary treatment stages for the domestic effluent. A comprehensive analysis over three years was then conducted to provide a robust characterization of the internal dynamics of the systems, particularly with respect to N and P removal as well as evaluating the temporal water balance across the different seasons. The removal of Total N was only 29% and 30% in the secondary and tertiary treatment wetlands, respectively; particularly disappointing for the tertiary treatment process, which was receiving nitrified effluent. Studies on the (15)N stable isotope confirmed that 35% of the ammonium from the septic tank was passing straight through the process without taking part in any biogeochemical processes. However, influent N in the wetlands was shown to be biologically assimilated into organic nitrogen and then released again as soluble ammonium--so-called nitrogen "spiraling." Removal of Total P in the wetlands averaged from 28% to 45% with higher P removals measured during summer periods, although the effluent concentrations were still found to be high (> 5 mg/l on average). The phosphorus in the plant material was also analysed revealing that the annual above-ground stem matter only accounted for 1.3% to 8.4% of the annual total P-load in the wetlands. Finally, the water balance analyses showed that the mean flow discharging from both the secondary and tertiary treatment wetlands was slightly greater than the mean flow to the reed bed over the trial period, with rainfall acting to increase flows by 13% and 5%, respectively, on average in winter while just about balancing evapotranspiration in the summer. PMID- 21644149 TI - Experimental study of bentonite-soil mixtures as anti-seepage materials of constructed wetlands. AB - In this study, mixtures of different kinds of bentonite and soil were used and tested in order to find a cheap alternative to current anti-seepage materials for constructed wetlands. The anti-seepage layer of constructed wetlands was simulated in the experimental study and the permeability coefficient of the mixed materials was determined in order to evaluate the anti-seepage effect of mixtures. The main results are as follows: (i) The minimum mass ratio of bentonite to soil is 10%; (ii) Within a certain range, the more compact and higher the wet density is, then the better anti-seepage effect is (under the condition of certain moisture content). The permeability coefficient of the mixed materials exponentially increased with the increase of wet density; (iii) At the wet density of 1.83 g/cm(3), corresponding with the optimum compactness, the mixture of natural sodium bentonite produced in Wyoming, USA and soil from Cangzhou, Hebei province showed the best anti-seepage performance; (iv) The impermeability of the mixture with smaller particle sizes of bentonite was far better than that with the bigger particle sizes; (v) The hydration effect of bentonite changed the structure of the mixture materials into a special structure that is similar to that of pure bentonite. The particles of the mixture became expanded under SEM investigation and the mixture became more compact, which could have the same or similar effect as pure bentonite for anti-seepage. PMID- 21644150 TI - Treatment of landfill leachate with high levels of ammonia by constructed wetland systems. AB - Landfill leachate treatment can be expensive and energy-intensive when achieved in advanced wastewater treatment plants. One possible method to reduce cost and energy requirements is to treat the leachate on-site using constructed wetlands. In this study, we investigated the feasibility of treating the leachate from a 10 year-old landfill site, with high levels of ammonia, by lab-scale constructed wetland microcosms. Five different types of wetland systems were used in the test: P1, P2 and CS-1 systems fed by original leachate, P3 and CS-2 systems fed by secondarily treated leachate. The CS systems were used as control without vegetation. Each system contained two basins connected in series: one installed as free water surface (FWS) wetland microcosm, and the other as subsurface flow (SSF) wetland microcosm. After one year of operation, the experimental results showed that P2 system performed better than both P1 and P3 systems by comparing their removal efficiencies of ammonia, which were measured equal to 94%, 84% and 60%, respectively. However, for total nitrogen removal, all three systems exhibited lower values, which meant that the process denitrification was insignificant in the wetland systems treating aged leachate. According to the experimental results of adding extra BOD into the systems, we found that the reason was due to lack of organic carbon sources in aged leachate for denitrification, which might cause nitrate buildup in the effluents. PMID- 21644151 TI - Nitrogen removal in an integrated constructed wetland treating domestic wastewater. AB - The nitrogen (N) removal performance of a 3.25 ha Integrated Constructed Wetland (ICW) treating domestic wastewater from Glaslough village in County Monaghan, Ireland, was evaluated in this study. The ICW consists of two sludge ponds and five shallow vegetated wetland cells. Influent and effluent concentrations of two N species, namely, ammonia-nitrogen (NH(3)-N) and nitrate-nitrogen (NO(3)-N), which were measured weekly over 2 years, together with hydrology of the ICW provided the basis for this evaluation. The influent wastewater typically contained 40 mg L(-1) NH(3)-N and 5 mg L(-1) NO(3)-N. Concentrations of N in the ICW effluent were typically less than 1.0 mg L(-1) for both species. Overall, a total load of 2802 kg NH(3)-N and 441 kg NO(3)-N was received by the ICW and a removal rate of 98.0 % and 96.9 %, respectively, was recorded. Average areal N loading rate (245 mg m(-2) d(-1) NH(3)-N and 38 mg m(-2) d(-1) NO(3)-N) had a significant linear relationship with areal N removal rate (240 mg m(-2) d(-1) and 35 mg m(-2) d(-1), respectively) for both species. The areal first-order N removal rate constants in the ICW averaged 14 m yr(-1) for NH(3)-N and 11 m yr( 1) for NO(3)-N. Temperature coefficients (theta) for N reduction in the ICW were lower and less than unity for NO(3)-N, suggesting that the variability in N removal by the ICW was marginally influenced by temperature. PMID- 21644152 TI - STELLA software as a tool for modelling phosphorus removal in a constructed wetland employing dewatered alum sludge as main substrate. AB - A dynamic simulation model was developed for the removal of soluble reactive phosphorus (SRP) from the vertical flow constructed wetlands (VFCW) using a dynamic software program called STELLA (structural thinking, experiential learning laboratory with animation) 9.1.3 to aid in simulating the environmental nature and succession of relationship between interdependent components and processes in the VFCW system. In particular, the VFCW employed dewatered alum sludge as its main substrate to enhance phosphorus (P) immobilization. Although computer modelling of P in treatment wetland has been well studied especially in recent years, there is still a need to develop simple and realistic models that can be used for investigating the dynamics of SRP in VFCWs. The state variables included in the model are dissolved phosphorus (DISP), plant phosphorus (PLAP), detritus phosphorus (DETP), plant biomass (PLBI) and adsorbed phosphorus (ADSP). The major P transformation processes considered in this study were adsorption, plant and microbial uptake and decomposition. The forcing functions which were considered in the model are temperature, radiation, volume of wastewater, P concentration, contact time, flow rate and the adsorbent (i.e., alum sludge). The model results revealed that up to 72% of the SRP can be removed through adsorption process whereas the uptake by plants is about 20% and the remaining processes such as microbial P utilization and decomposition, accounted for 7% SRP removal based on the mass balance calculations. The results obtained indicate that the model can be used to simulate outflow SRP concentration, and it can also be used to estimate the amount of P removed by individual processes in the VFCW using alum-sludge as a substrate. PMID- 21644153 TI - Towards the development of a novel construction solid waste (CSW) based constructed wetland system for tertiary treatment of secondary sewage effluents. AB - This study was conducted to examine the possibility of using construction solid waste (CSW), an inevitable by-product of the construction and demolition process, as the main substrate in a laboratory scale multi-stage constructed wetland system (CWs) to improve phosphorus (P) removal from secondary sewage effluent. A tidal-flow operation strategy was employed to enhance the wetland aeration. This will stimulate aerobic biological processes and benefit the organic pollutants decomposition and nitrification process for ammoniacal-nitrogen (NH(+)(4)-N) removal. The results showed that the average P concentration in the secondary sewage effluent was reduced from 1.90 mg-P/L to 0.04 mg-P/L. CSW presents excellent P removal performance. The average NH(+)(4)-N concentration was reduced from 9.94 mg-N/L to 1.0 mg-N/L through nitrification in the system. The concentration of resultant nitrite and nitrate in the effluent of the CSW based CWs ranged from 0.1 to 2.4 mg-N/L and 0.01 to 0.8 mg-N/L, respectively. The outcome of this study has shown that CSW can be successfully used to act as main substrate in CWs. The application of CSW based CWs on improving N and P removals from secondary sewage effluent presents a win-win scenario. Such the reuse of CSW will benefit both the CSW disposal and nutrient control from wastewater. More significantly, such the application can transfer the CSW from a 'waste' to 'useful' material and can ease the pressure of construction waste solid management. Meanwhile, the final effluent from the CSW-based CWs can be used as non-potable water source in landscape irrigation, agriculture and industrial process. PMID- 21644154 TI - Removal of Salmonella and indicator micro-organisms in integrated constructed wetlands treating agricultural wastewater. AB - The purpose of this study was to investigate the removal of pathogenic and indicator micro-organisms in integrated constructed wetland (ICW) systems treating agricultural wastewater. Nine ICW's treating piggery (3) or dairy (6) wastewaters were sampled and indicator micro-organisms were enumerated in the influent as well as the effluent from the first, mid- and final cells. The presence/absence of Salmonella was also determined and any Salmonella isolates recovered were characterized. Mean counts of coliform, E. coli and Enterococcus across all nine ICW systems were lower in the final effluent than in the effluent from cell 1 (P < 0.001). An antibiotic susceptible isolate of Salmonella Dublin, a bovine-adapted serotype, was isolated from the influent to one dairy ICW but was not detected in any of the ICW cells. An antibiotic sensitive Salmonella Dublin isolate with the same molecular fingerprint was also recovered from the cell 1 effluent of another dairy ICW but was absent from the influent and the mid cell and final effluents. Salmonella Typhimurium DT104b was detected in the liquid fraction of anaerobically digested pig manure as well as in the effluent from the first cell and mid-cell of an ICW treating this material, but was absent in the final effluent. Molecular fingerprinting by pulsed field gel electrophoresis demonstrated that the recovered isolates were highly related. However, they had different antimicrobial resistance profiles, with some highly resistant isolates recovered. In conclusion, counts of indicator micro-organisms were reduced significantly within ICW, with E. coli and Enterococcus non detectable in the final effluent. Moreover, Salmonella, when present in the influent, appears to have been removed. PMID- 21644155 TI - Treating surface water with low nutrients concentration by mixed substrates constructed wetlands. AB - Constructed wetland (CW) has been widely applied in nutrients reduction for eutrophication control, especially in the advanced treatment of effluent of municipal sewage plants or the in-lake river treatment with high hydraulic loads and low nutrient concentrations. But in real application, it shows lower nutrient removal efficiency. The main reason is that traditional substrates, such as soil and gravel have low capacity for nitrogen and phosphorus removal. This study aims to enhance nutrients removal in constructed wetland systems by using series of substrates including calcium silicate hydrate (CSH), vermiculite and ceramsite which are all investigated individually in static experiment or mixed in batch and continuous flow experiments. The result showed that the efficiency of phosphorus removal by CSH could reach 97%, much higher than the other substrates. However, when it was applied in CW, the removal efficiency decreased. Although vermiculite showed the highest ammonia nitrogen removal efficiency of 65.91%, the ammonia nitrogen removal efficiency may have depended on the action of microorganism. High total nitrogen removal efficiency was obtained in continuous flow mixed substrate CW. Under a hydraulic retention time (HRT) of 18h and hydraulic loading rate (HLR) of 0.496 m(3)/m(3).d, average total nitrogen removal efficiency of above 91% was achieved, but the average phosphorus removal efficiency was around 65% and this needs to be improved further. PMID- 21644156 TI - Effects of the substrate depth on purification performance of a hybrid constructed wetland treating domestic sewage. AB - The depth of substrate in constructed wetlands (CWs) has a significant effect on the construction investment and the purification performance of CWs. In this study, a pilot scale CW system was operated in a domestic sewage treatment plant in Xi'an, China. The experimental systems included three-series CWs systems with substrate depths of 0.1m, 0.3 m and 0.6 m, respectively. Each series was composed of a hydroponic ditch, a horizontal subsurface flow CW and a vertical flow CW. The effluent from the primary clarifier in the sewage treatment plant was intermittently conducted to the wetlands at a flow rate of 0.3 m(3)/d. The hydraulic loading rate of each CWs system was regulated at 0.1 m(3)/m(2).d and the hydraulic retention time was 3 days. Canna indica L. was planted both in the hydroponic ditches and the CWs systems. Results showed that the highest removal efficiency of NH(+)(4)-N and TP was obtained in the hybrid CW with 0.1 m substrate depth. The average removal efficiency for NH(+)(4)-N and TP were 90.6 % and 80.0 %, respectively. The highest average removal efficiency of COD was obtained in hybrid CWs system with 0.6 m substrate depth. Therefore, a simultaneous removal of COD and nutrients can be achieved through the combination of different wetlands using different substrate depths. In addition, the substrate depth presents significant effects on the concentration of DO and root growth characteristics of canna in the system. As a result, the highest concentration of DO (>2 mg/L) and the highest amount of roots production were achieved in the 0.1 m substrate depth horizontal and vertical flow CWs. PMID- 21644157 TI - Meso-scale systems used for the examination of different integrated constructed wetland operations. AB - Meso-scale constructed wetlands have not been commonly used for the examination of interactions and operations within differently designed wetland systems. Sixteen meso-scale constructed wetland systems (4 operations with 4 replicates each) were therefore evaluated between November 2008 and June 2010. These systems were used to examine key operations identified in the literature including hydraulic loading rates, nutrient loading rates and nutrient recycling modes. The wetlands were managed in the following modes: normal, recycling, high nutrient loading and high flow rate. The designs were such that the influents could be varied at set loading rates and concentrations. The ammonia removal rates for the meso-scale integrated constructed wetland (ICW) systems were similar to other full-scale ICW systems that have been in operation in the south-east of Ireland for the past decade. These comparable results highlight the potential cost effectiveness of these meso-scale experimental designs for the examination of novel operations for future ICW developments. PMID- 21644158 TI - Removal efficiency and balance of nitrogen in a recirculating aquaculture system integrated with constructed wetlands. AB - The nitrogen (N) balance for aquaculture is an important aspect, especially in China, and it is attributed to the eutrophication in many freshwater bodies. In recent years, constructed wetlands (CWs) have been widely used in wastewater treatment and ecosystem restoration. A recirculating aquaculture system (RAS) consisting of CWs and 4 fish ponds was set up in Wuhan, China. Channel catfish (Ictalurus punctatus) fingerlings were fed for satiation daily for 168 days with 2 diets containing 5.49 % and 6.53 % nitrogen, respectively. The objectives of this study were to investigate the N budget in the RAS, and try to find out the feasibility of controlling N accumulation in the fish pond. It is expected that the study can provide a mass balance for the fate of N in the eco-friendly treatment system to avoid eutrophication. The results showed that the removal rates of ammonia (NH(+)(4)-N), sum of nitrate & nitrite (NO(-)(X)-N), and total nitrogen (TN) by the CWs were 20-55%, 38-84 % and 39-57 %, respectively. Denitrification in the CWs was the main pathway of nitrogen loss (41.67 %). Nitrogen accumulation in pond water and sediment accounted for 3.39 % and 12.65 % of total nitrogen loss, respectively. The nitrogen removal efficiency and budget showed that the CW could be used to control excessive nitrogen accumulation in fish ponds. From the viewpoint of the nitrogen pollution control, the RAS combined with the constructed wetland can be applied to ensure the sustainable development for aquaculture. PMID- 21644159 TI - Application of constructed wetland for urban lake water purification: trial of Xing-qing Lake in Xi'an city, China. AB - A comprehensive review of the current water pollution status in China has indicated that the urban lakes in Chinese cities have suffered from serious pollution and are in high risk of eutrophication, although the pollution sources have been largely controlled. The objective of this study lies in exploring a long term restoration of the aquatic ecosystem in Chinese city lakes using treatment wetland, an environmentally friendly and cost-effective technology. Trials from a subsurface horizontal flow constructed wetland (CW) have demonstrated that the treatment wetland can be used for a purpose such as lake water quality control. Average removal of 84.2% for COD, 53.8% for NH(3)-N, 47.9% for TN, 73.3% for TP and 86.6% for SS can be achieved. Relatively, low removal of nitrogen lies in the lack of nitrification and denitrification process. Accordingly, improved configuration of the treatment wetland system has been proposed and discussed. Finally, the importance of the integrated constructed wetland especially for the application of urban lake water treatment is highlighted. PMID- 21644160 TI - Treating swine wastewater by integrating earthworms into constructed wetlands. AB - The aim of this study was to investigate the application of integrating earthworms (Pheretima peguana) into two-stage pilot-scale subsurface-flow constructed wetlands (SFCWs) receiving swine wastewater in terms of their treatment performance, namely organic content, total kjeldahl nitrogen (TKN), and solid reduction as well as the quantity of sludge production. There was a minor difference in terms of removal efficiency according to each parameter when comparing the unit with earthworms to the one without earthworms. Both achieved the TKN, biochemical oxygen demand (BOD), chemical oxygen demand (COD), total volatile suspended solids (TVSS), suspended solids (SS), and total solids (TS) removal by more than 90 %. The earthworms helped in reducing the sludge production on the surface of constructed wetlands 40 % by volume, which resulted in lowering operational costs required to empty and treat the sludge. The plant biomass production was higher in the wetlands without earthworms. Further research could be undertaken in order to effectively apply earthworms inside the wetlands. PMID- 21644161 TI - Major and trace element distribution in the peat from ombrotrophic bogs in Latvia. AB - This study was undertaken to analyse major and trace elements and the nature of their accumulation in peat, with a particular emphasis on peat properties and the impact of local and regional pollution sources on the character of element accumulation in ombrotrophic bogs in Latvia. The element concentration values in peat from Latvia reflect the local processes that affect element concentrations in the peat mass, indicating accumulation of trace elements - apparently of anthropogenic origin (Pb, Cd, Co, Ni and others) - in the upper layers of the peat profiles. In addition, they indicate accumulation of several elements (for example, As, Cr and others) in deeper layers of bog, possibly due to the feeding pattern, depending on the saturation of the groundwater. PMID- 21644162 TI - Acyclic monoterpenes in tree essential oils as a shrinking agent for waste expanded polystyrene. AB - We examined the dissolution of polystyrene (PS) into acyclic monoterpenes present in tree essential oils, to develop an environmentally friendly shrinking agent for waste-expanded polystyrene (EPS). The dissolving powers of geranyl acetate, geranylacetone, and geranyl formate [221.8-241.2 g PS (100 g solvent)(-1)] compared favorably with that of (R)-limonene [181.7 g PS (100 g solvent)(-1)]. Their favorable dissolving powers for PS can be explained by their flexible linear structures, which may be more accessible to the inside of bulk PS compared with cyclic monoterpenes. These acyclic monoterpenes and PS were recovered almost quantitatively by simple steam distillation of the PS solution. PMID- 21644163 TI - Health risk assessment of inhabitants exposed to PAHs particulate matter in air. AB - The presence of polycyclic aromatic hydrocarbon compounds (PAHs) was investigated in the particulate matter of samples taken at six sampling sites in the city of Curitiba (southern Brazil). The concentrations of suspended particulate matter ranged from 11.02 to 177.27 ng/m(3). The analysis showed that 14 of the 16 PAHs are considered a priority compound by the USEPA (US Environmental Protection Agency). The mixture of PAHs was predominantly composed of PAHs with 3 and 4 aromatic rings. Isomer pair ratios show that the main source of PAHs was from burning fossil fuels (gasoline and diesel oil), although other sources may have contributed also. Benzo(a)pyrene, (BaP), regarded one of the most toxic PAHs, was present in all samples, but with concentrations lower than the maximum concentrations defined by some EU Countries. The risk assessment was conducted using the toxic equivalent factor (TEF) considering the toxicity of the individual PAHs compared to BaP. The BaP(eq) for all samples was between 0.42 1.12 ng/m(3). The equivalent BaP(eq) indicated low health risk associated with exposure to the total PAHs content in air. The incremental lifetime cancer risk (ILCR) model was used to find the risk level for workers close to the emitting pollution sources. Outdoor exposure showed no risk for the amount of PAHs emitted. The acceptable risk is 10(-6); however, all results were lower than this value. PMID- 21644164 TI - The use of cork waste as a biosorbent for persistent organic pollutants-Study of adsorption/desorption of polycyclic aromatic hydrocarbons. AB - The aim of this study is to determine the sorption-desorption behavior of a mixture of thirteen aqueous PAHs on cork waste at a particle of size 0.25-0.42 mm obtained from the remains of cork strips. The final purpose is to use this natural adsorbent as an alternative to activated carbon in an innovative approach for the removal of this class of toxic compounds, and significantly reduce the regeneration costs of the process. The chemical composition of the selected cork revealed that suberin (38.5 %) and lignin (31.6 %) were the main structural components of the cell wall. The high efficiency of cork as a biosorbent of PAHs is shown by the fact that just over 80 % of adsorption occurred during the first two minutes of contact time. Both Freundlich's and Langmuir's isotherms gave good fits to the sorption process. The highest adsorption affinities were exhibited for pyrene, anthracene, and phenanthrene. Desorption studies indicate a high degree of irreversibility for all PAHs, and especially so in the case of high molecular PAHs. The correlation with K(F) and low molecular weight PAHs was the most significant. The quantity of cork required to reduce water pollution was estimated to be between 3 and 15 times less than the quantities required in the case of other materials (i.e. aspen wood and leonardite). This study demonstrates for the first time that cork is a potential biosorbent for PAHs and may have relevance in the future treatment of PAH-contaminated waters. PMID- 21644165 TI - Effluent monitoring at a bleached kraft mill: directions for best management practices for eliminating effects on fish reproduction. AB - A long-term monitoring study was conducted on effluents from a bleached kraft pulp and paper mill located in Eastern Canada. The study was designed to gain insights into temporal effluent variability with respect to fish reproduction as it related to production upsets, mill restarts and conditions affecting biological treatment performance. Final effluent quality was monitored between February 2007 and May 2009 using biochemical and chemical oxygen demand, total suspended solids, resin and fatty acids, a gas chromatographic profiling index, and the presence of methyl substituted 2-cyclopentenones. Selected effluent samples were evaluated for effects on fish reproduction (egg production) using a shortened version of the adult fathead minnow reproductive test. The events relating to negative effects on fish reproduction were upsets of the pulping liquor recovery system resulting in black liquor losses, operational upsets of the hardwood line resulting in the loss of oxygen delignification filtrates, and conditions that reduced the performance of biological treatment (e.g., mill shutdown and low ambient temperatures). The reductions in egg production observed in fathead minnow were associated with biochemical oxygen demand values > 20 mg/L, GC profiling indices > 1.2 and the presence of methyl-substituted 2 cyclopentenones at concentrations > 100 MUg/L. This study demonstrated the importance of both in-plant measures for controlling the loss of organics as well as the optimum operation of biological effluent treatment for eliminating effluent-related effects on fish reproduction (egg production) in the laboratory. PMID- 21644166 TI - Individual differences in motivational activation influence responses to pictures of taboo products. AB - In this article, the authors investigated responses to pictures of products whose use is socially or legally restricted for teens and young adults (e.g., beer, liquor, cigarettes). The authors theorized and found that these pictures are motivationally relevant and therefore elicit automatic activation in the appetitive/approach or aversive/defensive motivational systems, which leads to increased attention, arousal, emotional response, and memory for the risky products. The authors also found that these responses are mediated by individual differences in motivational reactivity. The authors suggest that placing images of these products in prevention messages may work against the prevention goal by increasing appetitive activation and positive emotion in populations more inclined to take risks. PMID- 21644167 TI - Spousal communication about HIV prevention in Kenya. AB - High HIV rates among cohabiting couples in many African countries have led to greater programmatic emphasis on spousal communication in HIV prevention. This study examines how demographic and socioeconomic characteristics of cohabiting adults influence their dyadic communication about HIV. A central focus of this research is on how the position of women relative to their male partners influences spousal communication about HIV prevention. The authors analyze gaps in spousal age and education and females' participation in household decision making as key factors influencing spousal communication about HIV, while controlling for sexual behaviors of both partners as well as other individual and contextual factors. Data were obtained from the 2003 Kenya Demographic and Health Survey for 1,388 cohabiting couples. Information regarding spousal communication was self-reported, assessing whether both, either, or neither partner ever discussed HIV prevention with the other. Analyses showed higher levels of education for the female partner and participation in household decision making are positively associated with spousal communication about HIV prevention. With females' education and other factors controlled, couples with more educated male partners were more likely to have discussed HIV prevention than couples in which both partners have the same level of education. Spousal communication was also positively associated with household wealth status and exposure to the mass media, but couples in which male partners reported having nonspousal sex in the past year were less likely to have discussed HIV prevention with their spouses. Findings suggest HIV prevention programs should promote female empowerment and encourage male participation in sexual health discussion. PMID- 21644168 TI - Athletic identity and disordered eating in obligatory and non-obligatory runners. AB - Athletic identity is the extent to which an individual identifies with being an athlete. Strong "running" role identity may contribute to increased restrictive dieting behaviours, potentially placing such individuals at risk for eating disorders. In this study, we examined differences in eating and exercise behaviours/attitudes and athletic identity in obligatory versus non-obligatory runners. Male and female participants completed a battery of questionnaires including the Eating Disorder Inventory, Obligatory Exercise Questionnaire (OEQ), and Athletic Identity Measurement Scale (AIMS). OEQ scores >=50 were indicative of obligatory exercise. The non-obligatory runners (n = 82) and obligatory runners (n = 91) were compared on the various measures. Obligatory runners scored significantly higher (P < 0.002) on all of the eating attitudes/disorder measures, and the AIMS (P <= 0.006). Scores on the AIMS were correlated with all disordered eating measures (P < 0.05). Exercising to maintain identification with the running role may be associated with pathological eating and training practices. PMID- 21644169 TI - Anti-inflammatory and antioxidant activity of Ficus carica Linn. leaves. AB - Ficus carica Linn. (Moraceae) is commonly known as edible fig. The leaves, roots, fruits and latex of the plant are medicinally used in different diseases. The leaves are claimed to be effective in various inflammatory conditions like painful or swollen piles, insect sting and bites. However, there has been no report on anti-inflammatory and antioxidant activity of F. carica leaves. Therefore the aim of this study was to evaluate the anti-inflammatory and antioxidant activity of F. carica leaves. Our study validated the traditional claim with pharmacological data. Anti-inflammatory and antioxidant activity of the drug could be due to the presence of steroids and flavanoids, respectively, which are reported to be present in the drug. Furthermore, the anti-inflammatory activity of the drug could be due to its free radical scavenging activity. Further work is also required to isolate and characterise the active constituents responsible for the anti-inflammatory activities. PMID- 21644170 TI - Studies on the potential antioxidant properties of Senecio stabianus Lacaita (Asteraceae) and its inhibitory activity against carbohydrate-hydrolysing enzymes. AB - This study showed for the first time the antioxidant and hypoglycaemic properties of the methanol, n-hexane and ethyl acetate extracts from Senecio stabianus Lacaita, a plant that belongs to the Asteraceae family. The antioxidant activities were carried out using two different in vitro assays, namely 2,2 diphenyl-1-picrylhydrazyl (DPPH) test and 2,2'-azinobis(3-ethylbenzthiazoline-6 sulphonate) (ABTS) test. The ethyl acetate extract showed the highest activity with inhibitory concentration 50% (IC(50)) values of 35.5 and 32.7 ug mL(-1) on DPPH test and ABTS test, respectively. This activity may be related to a good total phenol and flavonoid content. All extracts were also tested for their potential inhibitory activity of alpha-amylase and alpha-glucosidase digestive enzymes. The n-hexane extract exhibited the highest alpha-amylase inhibition with an IC(50) value of 0.21 mg mL(-1). Through bioassay-guided fractionation processes seven fractions (A-G) were obtained and tested. Based on the phytochemical analysis, the activity of n-hexane extract may be related to the presence of monoterpenes and sesquiterpenes. PMID- 21644171 TI - Influence of macro elements and nitrogen source on adventitious root growth and withanolide-A production in Withania somnifera (L.) Dunal. AB - Withania somnifera (L.) Dunal. (Indian ginseng) is an important medicinal plant which yields pharmaceutically active compounds, namely withanolides. This study deals with the optimisation of the adventitious root suspension culture of W. somnifera for the production of biomass and withanolide-A. We investigated the effects of macro elements (NH(4)NO(3), KNO(3), CaCl(2), MgSO(4) and KH(2)PO(4)) and nitrogen source [[Formula: see text]] of Murashige and Skoog (MS) medium on the accumulation of biomass and withanolide-A content. The highest accumulation of fresh and dry biomass (127.52 and 12.45 g L(-1)) was recorded in the medium with 0.5* concentration of NH(4)NO(3) and the highest production of withanolide-A was recorded in the medium with 2.0* KNO(3) (14.00 mg g(-1) DW). The adventitious root growth was greater when the [Formula: see text] concentration was higher than that of [Formula: see text] and the withanolide-A production was highest in the absence of [Formula: see text]. Maximum biomass growth was achieved at [Formula: see text] ratio of 14.38 : 37.60, while withanolide-A production was greatest (11.76 mg g(-1) DW) when the [Formula: see text] ratio was 0.00 : 18.80 mM. The results of this study are useful for scale-up processes. PMID- 21644172 TI - A new alkaloid from the marine-derived fungus Hypocrea virens. AB - A new alkaloid, 2-methylimidazo[1,5-b]isoquinoline-1,3,5(2H)-trione, was isolated from the EtOAc extract of the marine-derived fungus Hypocrea virens. Its structure was confirmed by spectroscopic methods (1D and 2D NMR, MS). PMID- 21644173 TI - Determination of the chemical composition and in vitro antioxidant activities of essential oil and methanol extracts of Echinophora platyloba DC. AB - This study was designed to examine the chemical composition and in vitro antioxidant activity of essential oil and methanol extracts of Echinophora platyloba from Iran. Gas chromatography (GC) and GC/MS (mass spectrometry; MS) analysis of the essential oil resulted in the identification of 29 compounds, which comprised 97.4% of the oil. The main constituents were found to be: (Z) beta-ocimene (26.7%), Delta-3-carene (16.2%) and limonene (6.6%). Antioxidant activities of the essential oil and the methanolic extracts from E. platyloba were evaluated using three different test systems, namely 2,2-diphenyl-1 picrylhydrazyl (DPPH) radical scavenging, beta-carotene-linoleic acid bleaching and reducing power assays. In the DPPH system, the highest radical-scavenging activity was shown by the polar sub-fraction of methanol extract (71.2 +/- 1.11 ug mL(-1)). Also in the second case, the relative inhibition capacity (%) of the essential oil (68.0 +/- 1.14%) was found to be the stronger one. In addition, the amounts of total phenol components in the polar sub-fractions of methanolic extract (67.5 +/- 0.48 ug mg(-1)), nonpolar sub-fractions of methanol extract (35.3 +/- 0.12) and the oil (83.3 +/- 0.24 ug mg(-1)) were determined. PMID- 21644174 TI - Leaf volatiles and secretory cells of Alpinia zerumbet (Pers.) Burtt et Smith (Zingiberaceae). AB - Plant leaves are commonly used in folk medicine and food industry. Their volatile composition is an important determinant in such applications. However, to properly assess the quality of volatiles, proper analytic tools must be utilised. Accordingly, the static headspace technique was used to evaluate the main volatiles emitted from in vitro-grown Alpinia zerumbet plants cultured with indole-3-acetic acid, thidiazuron, benzyladenine or kinetin, under standard physical conditions, as compared to those of field-grown donor plants. Although the leaf aroma of the donor plants was found to be a complex mixture, mainly consisting of sabinene, alpha and gamma-terpinene, 1,8-cineole and caryophyllene, volatile analyses from most of the in vitro samples only revealed the presence of sabinene and caryophyllene. Many alkanes were found in the aromas after treating plantlets with cytokinins. Histochemical analysis of leaf sections was also carried out. Secretory cells found in the epidermis and mesophyll showed a strong positive reaction to lipophilic compounds using Oil red and Nile blue reagents. These findings demonstrated how in vitro conditions may alter the quality of volatiles in micropropagation systems, while leaf anatomy analysis revealed a large quantity of oil cells in the mesophyll as a constant feature responsible for the production of volatile compounds in both donor and in vitro-grown plants. PMID- 21644175 TI - Chemical constituents and in vitro antioxidant activity of Phyllanthus wightianus. AB - The whole plant of Phyllanthus wightianus (PW) was investigated for the antioxidant effects of three successive extracts: hexane (PWHE), chloroform (PWCE) and methanol (PWME), using standard in vitro models. The PWME exhibited a strong scavenging effect on 2,2-diphenyl-2-picryl hydrazyl (DPPH) free radicals and nitric oxide radical inhibition activity, due to possessing the highest content of tannins. The free radical scavenging effect of PWME was comparable with that of reference antioxidants. The extracts were subjected to isolation of their compounds: isomeric sterol mixture (1) [stigmasterol (1a), compesterol (1b) and beta-sitosterol (1c)], fredilin (2), lupeol (3), gallic acid (4), bergenin (5), geraniin (6), corilagin (7) and ellagic acid (8) were established through the use of column chromatographic methods and spectral data. The percentage of tannins was also determined and estimated using the HPLC method. The data suggest that tannins are the active antioxidant compounds of P. wightianus. This study provides proof for the ethnomedical claims and reported biological activities of this plant. The plant therefore has very good therapeutic potential. PMID- 21644176 TI - Pregnane and pregnane tetraglycoside from Marsdenia roylei. AB - Pregnane oligoglycoside, namely roylinine, and a pregnane derivative, namely marsgenin, have been isolated from chloroform-soluble extract of dried stem of Marsdenia roylei, and their structures were determined using 1H-NMR, 13C-NMR, 1H 1H COSY, HSQC, TOCSY and FABMS spectral techniques as well as chemical degradation and derivatisation. PMID- 21644177 TI - Extraction of kiwi seed oil: Soxhlet versus four different non-conventional techniques. AB - Kiwi seed oil has a nutritionally interesting fatty acid profile, but a rather low oxidative stability, which requires careful extraction procedures and adequate packaging and storage. For these reasons and with the aim to achieve process intensification with shorter extraction time, lower energy consumption and higher yields, four different non-conventional techniques were experimented. Kiwi seeds were extracted in hexane using classic Soxhlet as well as under power ultrasound (US), microwaves (MWs; closed vessel) and MW-integrated Soxhlet. Supercritical CO2 was also employed and compared to the other techniques in term of yield, extraction time, fatty acid profiles and organoleptic properties. All these non-conventional techniques are fast, effective and safe. A sensory evaluation test showed the presence of off-flavours in oil samples extracted by Soxhlet and US, an indicator of partial degradation. PMID- 21644178 TI - Chemical compositions of the rhizome oils of two Alpinia species of Malaysia. AB - The essential oils obtained by hydrodistillation of the rhizomes of Alpinia aquatica Rosc. syn. Alpinia melanocarpa and Alpinia malaccensis Roscoe were analysed by capillary gas chromatography and gas chromatography-mass spectrometry. Eighteen compounds, representing 98.4% of the essential oil were identified in A. aquatica rhizome oil, with beta-sesquiphellandrene in 36.5% being the major constituent, while 20 compounds representing 99.7% of the rhizome oil of A. malaccensis were identified, among which methyl (E)-cinnamate (78.2%) was the major constituent. PMID- 21644179 TI - Two new special flavones from Uvaria accuminata. AB - From the stem bark of Uvaria accuminata two new compounds, 5-methoxy-6,8,8 trimethyl-2-phenylchromene-4,7-dione (1) and a sesquiterpene-fused flavone, named acuminavone (2), were isolated together with a known flavone, desmosdomutin or dasytrichone, (5-hydroxy-6,8,8-trimethyl-2-phenylchromene-4,7-dione). The structure elucidation of compounds 1 and 2 in spectroscopic studies is described. PMID- 21644180 TI - Artomandin, a new xanthone from Artocarpus kemando (Moraceae). AB - A new furanodihydrobenzoxanthone, artomandin (1), together with three other flavonoid derivatives, artoindonesianin C, artonol B, and artochamin A, as well as beta-sitosterol were isolated from the stem bark of Artocarpus kemando. The structures of these compounds were determined on the basis of spectral evidence. All of these compounds displayed inhibition effects to a very susceptible degree in cancer cell line tests. Compound 1 also exhibited significant antioxidant capacity in the free radical 1,1-diphenyl-2-picrylhydrazyl tests. PMID- 21644181 TI - Volatile constituents of Zosimia radians Boiss. and Hohen from Iran. AB - The essential oil of the aerial parts of Zosimia radians Boiss. & Hohen belonging to the Apiaceae family was obtained by hydrodistillation and analysed by GC and GC-MS. Thirty-nine compounds representing 95.7% of the oil were identified. Among them, citronellyl acetate (16.3%) and octyl butyrate (15.0%) were the most abundant. PMID- 21644182 TI - Secondary metabolites of Pyrenochaeta sp. B36, an endophytic fungus from Annona squamosa L. AB - Four new metabolites, named pyrenochaetamide A, pyrenochaetolide A, pyrenochaetoxy A and pyrenochaetolide B, together with five known compounds, were isolated from strain Pyrenochaeta sp. B36, an endophytic fungus of Annona squamosa L. Their structures were elucidated by spectroscopic analyses including 1D- and 2D-NMR (HMQC, HMBC, 1H-1H COSY and NOESY) and HRFT-MS. The cytotoxic activities of these compounds were evaluated and some of them showed substantial effect. PMID- 21644183 TI - Isolation of two major iridoid glucosides from Incarvillea emodi. AB - Two major iridoid glucosides, namely plantarenaloside, a neurotrophic compound, and boschnaloside, an antibacterial iridoid glucoside, have been isolated from the shoots of Incarvillea emodi (a wild ornamental plant). The compounds are characterised by NMR spectral data and ESI mass spectrometery. This is the first ever report on the isolation of iridoid glucosides from I. emodi (Bignoniaceae). PMID- 21644184 TI - Development of an intervention to reduce transmission of respiratory infections and pandemic flu: measuring and predicting hand-washing intentions. AB - This was an exploratory pilot study forming part of a programme of work to develop and trial an effective web-based intervention to reduce the risk of transmission of respiratory infections by promoting hand washing and other preventive behaviours in pandemic and non-pandemic contexts. The main purpose of this study was to confirm that the behavioural determinants we had identified from theory were related as predicted to intentions and to establish the validity of our measures of behavioural intentions. Participants (N = 84) completed a self report web-delivered questionnaire measuring intentions to engage in hand washing and the hypothesised behavioural determinants of intentions, based on the theory of planned behaviour and protection motivation theory. In a factorial 2 * 2 design, half of the participants were first randomised to receive messages about potential negative consequences of pandemic flu (the "high-threat" condition) and half were assigned to receive "coping" messages describing the rationale and effectiveness of hand washing for reducing the risk of infection. A substantial proportion of variance in intentions was explained by measures of attitudes (instrumental and affective), social norms (descriptive and injunctive), perceived behavioural control (especially, access to hand gel) and perceived risk (in particular, the likelihood of catching pandemic flu). Our measures of intentions were sensitive to between-group differences, and although our design did not permit causal inference (particularly in view of selective dropout among those required to read most web pages), the pattern of differences was in the expected direction, that is, hand-washing intentions tended to be stronger in those receiving the high-threat message and coping messages. This study provided encouraging confirmation that our intervention development was proceeding correctly. Measures of intentions proved sensitive to group differences, and the behavioural determinants included in the study explained a substantial proportion of the variance in intentions. The study also provided useful indications that our high-threat message might increase hand-washing intentions, that providing hand gel might be beneficial and that it would be necessary to actively manage the risk of selective dropout in the intervention group. PMID- 21644185 TI - Why are some people with neurological illness more resilient than others? AB - The current qualitative study was designed to evaluate the coping strategies of people living with a chronic progressive neurological illness and their carers. The neurological illnesses were Huntington's disease, motor neurone disease, multiple sclerosis and Parkinson's disease. Participants included 15 people who showed high levels of adjustment and 15 who showed low levels of adjustment. Participants were selected from an earlier study, to ensure that they satisfied the inclusion criteria for the current study. Interviews were completed to determine the strategies used to cope with the demands of the illness. Participants who demonstrated good adjustment were more likely to draw on social support to provide them with the resources to deal with the illness. In contrast, those who evidenced poor adjustment were more likely to draw on external supports to complete tasks for them. The implications of these findings for people with chronic neurological illnesses and their families are discussed. PMID- 21644186 TI - Brief strategic therapy in patients with social phobia with or without personality disorder. AB - The aim of this study was to test the effects of brief strategic therapy in people with social phobia distinguished by the presence or absence of DSM anxious personality disorder. A group of 120 outpatients with social phobia, 60 of them with comorbid DSM anxious personality disorder, were randomly assigned to 10 weeks of brief strategic therapy or minimal supportive therapy. Outcome analyses were conducted on the intent-to-treat sample at posttreatment and 3-month follow up using self-report interpersonal sensitivity and phobic anxiety measures, and at 6-month follow-up using social phobia diagnostic status. Brief strategic therapy was superior to minimal supportive therapy in patients with social phobia only. It was not significantly better in patients with comorbid personality disorder. It was concluded that 10 weeks of brief strategic therapy shows promise as a cost-effective method for management of social phobia. It has limited value as a method for management of social phobia with personality disorder. PMID- 21644187 TI - Coming to terms. AB - The assimilation model is a theory of psychological change that depicts the self as a community of internal voices, composed of traces of the person's experiences. The model suggests that disconnection of certain voices from the community underlies many forms of psychopathology and psychological distress. Such problematic voices may be assimilated through psychotherapeutic dialogue by building meaning bridges. Meaning bridges are signs (e.g., words, images, gestures, narratives) that have similar meaning to author and addressee, that is, to the signs' producer and recipient, which may be different people or interacting internal voices. Building meaning bridges is thus a process of coming to terms with problematic voices, which reduces distress and gives access to experiential resources within the self. This article describes and illustrates meaning bridges, voices, signs, and associated concepts as elaborated in a program of research on the assimilation model. PMID- 21644188 TI - Client experiences of motivational interviewing for generalized anxiety disorder: a qualitative analysis. AB - While Motivational Interviewing (MI) has demonstrated efficacy, little is known about the mechanisms through which MI achieves beneficial effects or how clients perceive the process of MI. The present study addressed this gap through a qualitative analysis of client accounts following four sessions of MI for generalized anxiety disorder. Clients identified increased motivation for treatment and change, experiencing the therapist as empathic and MI as a safe place to explore their feelings regarding change. MI was also described as deviant from client initial expectations. Overall, the emergent understanding of MI derived from clients' post-treatment narratives was consistent with MI principles and processes. PMID- 21644189 TI - Semantic size does not matter: "bigger" words are not recognized faster. AB - Sereno, O'Donnell, and Sereno (2009) reported that words are recognized faster in a lexical decision task when their referents are physically large than when they are small, suggesting that "semantic size" might be an important variable that should be considered in visual word recognition research and modelling. We sought to replicate their size effect, but failed to find a significant latency advantage in lexical decision for "big" words (cf. "small" words), even though we used the same word stimuli as Sereno et al. and had almost three times as many subjects. We also examined existing data from visual word recognition megastudies (e.g., English Lexicon Project) and found that semantic size is not a significant predictor of lexical decision performance after controlling for the standard lexical variables. In summary, the null results from our lab experiment--despite a much larger subject sample size than Sereno et al.--converged with our analysis of megastudy lexical decision performance, leading us to conclude that semantic size does not matter for word recognition. Discussion focuses on why semantic size (unlike some other semantic variables) is unlikely to play a role in lexical decision. PMID- 21644190 TI - Integrating working memory capacity and context-processing views of cognitive control. AB - Individuals low in working memory capacity (WMC) exhibit impaired performance on a variety of cognitive control tasks. The executive-attention theory of WMC (Engle & Kane, 2004) accounts for these findings as failures of goal maintenance and response conflict resolution. Similarly, the context-processing view (Braver et al., 2001) provides an explanation of cognitive control deficits observed in schizophrenia patients and older adults that is based on the ability to maintain context information. Instead of maintenance deficits, the inhibition view (Hasher, Lustig, & Zacks, 2007) states that older adults and individuals low in WMC primarily have an impairment in the ability to inhibit information. In the current experiment, we explored the relationships among these theories. Individuals differing in performance on complex span measures of WMC performed the AX-Continuous Performance Test to measure context-processing performance. High-WMC individuals were predicted to maintain the context afforded by the cue, whereas low-WMC individuals were predicted to fail to maintain the context information. Low-WMC individuals made more errors on AX and BX trials and were slower to respond correctly on AX, BX, and BY trials. The overall pattern of results is most consistent with both the executive-attention and context processing theories of cognitive control. PMID- 21644191 TI - Mortality in late nineteenth-century Montreal: geographic pathways of contagion. AB - In the City of Montreal, 1881, the presence of three cultural communities with different profiles of economic status makes it possible to observe the way social settings affected survival over a lifetime. Regression models show culturally determined maternal factors dominant for infants, and persistent throughout childhood. For post-neonates, children aged 1-4, and adults aged 15-59 household poverty has a comparable effect. Among adults, a gender penalty differs among the three communities. Models are improved when differentiated by cause of death. Locating households using a GIS reveals high levels of residential segregation by ethnicity and income, spatial correlation of environmental hazards, and constraints on exit from zones of risk, which together produce neighbourhood effects as large as household effects. Attention to groups excluded (foundlings and inmates of institutions) confirms that models limited to full household-level information significantly underestimate the impacts of poverty and exclusion. PMID- 21644192 TI - Imaging of non-neoplastic bone marrow disorders. PMID- 21644193 TI - Bone marrow changes in stress injuries. AB - Stress injuries and associated bone marrow changes are a common finding in athletes of all levels. Magnetic resonance imaging (MRI) is the imaging modality of choice for detecting characterizing and staging these injuries. However, because bone marrow edema patterns may also be seen in asymptomatic athletes, it is imperative for the sports medicine physician and the radiologist to closely correlate the imaging findings with the clinical signs and symptoms to assess the clinical significance of the imaging findings. This article reviews the pathophysiology, MRI findings, and clinical implications of stress injuries in athletes. PMID- 21644194 TI - Imaging of insufficiency fractures. AB - This review article focuses on occurrence, imaging, and differential diagnosis of insufficiency fractures. Prevalence and the most common sites of insufficiency fractures and their clinical implications are discussed. Insufficiency fractures are due to normal stress exerted on weakened bone. Most commonly postmenopausal osteoporosis is the cause for insufficiency fractures. Additional conditions affecting bone turnover include osteomalacia, chronic renal failure, and high dose corticosteroid therapy. It is a challenge for the radiologist to detect and diagnose insufficiency fractures as well as to differentiate them from malignant fractures. Radiographs are the basic modality used for screening of insufficiency fractures, yet depending on the location of the fractures, sensitivity is limited. Magnetic resonance imaging is a very sensitive tool to visualize bone marrow abnormalities associated with insufficiency fractures and allows differentiation of benign versus malignant fractures. Thin section multidetector computed tomography (CT) depicts subtle fracture lines allowing direct visualization of cortical and trabecular bone. Dedicated Mikro-CTs (Xtreme-CT) can detect subtle fractures reaching an in-plane resolution of 80 MUm. Bone scintigraphy still plays a role in detecting fractures, with good sensitivity but unsatisfactory specificity. Positron emission tomography-CT with hybrid-scanners has been the upcoming modality for the differentiation of benign from malignant fractures. Bone densitometry and clinical fracture history may determine the future risk of possible insufficiency fractures. PMID- 21644195 TI - Bone marrow edema pattern around the knee on magnetic resonance imaging excluding acute traumatic lesions. AB - Magnetic resonance imaging (MRI) is very sensitive for the detection of marrow abnormalities. Bone marrow edema on MRI has been defined as an area of low signal intensity on T1-weighted images, associated with intermediate or high signal intensity findings on T2-weighted images. The bone marrow edema pattern is a nonspecific finding with multiple etiologies. The knee is a common place for bone marrow signal abnormalities to appear on MRI. Besides contusions and fractures from acute trauma, there are a variety of other causes of the bone marrow edema pattern. It is important for the interpreter of the study to be aware of the different etiologies responsible for producing these changes and to be able to narrow the differential diagnosis without mistaking such a pattern for acute trauma or infiltrative tumor. This article concentrates on those entities that produce a bone marrow edema pattern not related to acute trauma including red marrow proliferation, stress, osteochondral lesions, osteonecrosis, bone marrow edema syndrome, arthropathy, infection, Paget's disease, and marrow replacement disorders. PMID- 21644196 TI - A practical guide to imaging of cartilage repair with emphasis on bone marrow changes. AB - Orthopedic surgeons have multiple options available to treat articular cartilage lesions, including microfracture, osteochondral autografting, and autologous chondrocyte implantation. By having basic knowledge of these surgical procedures, radiologists can more accurately interpret imaging studies obtained after surgery. In this article, we briefly review the different types of cartilage repair procedures, their appearance on magnetic resonance imaging (MRI), and pathologic MRI findings associated with postoperative complications. We also briefly discuss advanced MRI techniques (T2 mapping, delayed gadolinium-enhanced MRI of cartilage, sodium MRI) that have been recently used to assess the biochemical composition of repair tissue matrix. MRI can accurately assess the status and health of cartilage repair tissue. By providing this information to orthopedic surgeons, radiologists can play a valuable role in the management of patients who undergo cartilage repair surgery. PMID- 21644197 TI - Bone marrow changes in osteoarthritis. AB - Magnetic resonance imaging (MRI) is gaining increasing significance to study disease burden and progression in osteoarthritis. Bone marrow edema pattern has been identified as a typical MR finding associated with osteoarthritis. Histologically, bone marrow edema pattern is characterized by a mix of different pathologies, including bone marrow necrosis, fibrosis, microfractures, and bone remodeling as well as fibrovascular ingrowth. On MR imaging, bone marrow edema pattern is typically associated with subchondral cysts, cartilage defects, and meniscal degeneration; prevalence and size correlates with disease severity. Bone marrow edema pattern may be found in association with other abnormalities such as subchondral insufficiency fractures. Clinically the significance of bone marrow edema is controversially discussed with some studies showing evidence of associated pain. In summary, bone marrow signal abnormalities are frequent MRI signs of osteoarthritis with different histological findings, typical associated abnormalities, and also clinical significance. PMID- 21644198 TI - Dark t1 bone marrow. AB - Bone marrow contains fat and water. Using magnetic resonance (MR) T1-weighted images, fat appears light gray and water appears dark gray. Black or low signal structures include normal cortical bone, bone islands, hemosiderin deposits, calcification, ossification, metal artifact, gas, and foreign material. Reactivation of bone marrow reduces the signal of fatty marrow. Additional sequences and supplementary imaging with conventional radiographs and computed tomography are often helpful in determining the nature of the low signal lesions. Genetic bone disorders such as melorheostosis and other syndromes may give low signal lesion in a typical and often diagnostic pattern. MRI is an important method of showing the fracture line as a low signal lesion on T1-weighted images and is the preferred imaging method when conventional radiographs are unhelpful and fracture is suspected. Infection will cause edema and later sclerosis and necrosis with a mixed low and high signal pattern. Infarction will cause a mixed pattern depending on the stage of the process, but this includes low signal on T1 weighted images. Ossification and calcification in primary and secondary tumors appear as low signal areas on T1-weighted images. Recognition and understanding of lesions that cause low signal on T1-weighted images is an important tool when interpreting MR images. PMID- 21644199 TI - MRI evaluation of bone marrow changes in the diabetic foot: a practical approach. AB - One of the most important roles of magnetic resonance (MR) in imaging of the diabetic foot is to differentiate between the common and often comorbid pathologies that present with abnormal bone marrow signal. The primary diagnostic challenges in this setting are to distinguish osteomyelitis from reactive bone marrow edema, neuroarthropathy from osteomyelitis, and the sterile from the superinfected neuropathic joint. Whereas both osteomyelitis and reactive marrow edema share increased T2 signal, osteomyelitis is confirmed by T1 hypointensity in the bone marrow and reactive edema demonstrates isolated T2 signal hyperintensity. In distinguishing osteomyelitis from neuroarthropathy, a localized or contiguously spreading forefoot focus of abnormal bone marrow away from the subchondral surface and adjacent to a skin ulcer, cellulitis, abscess, or sinus tract would be indicative of osteomyelitis. A midfoot, subchondral, periarticular, or polyarticular distribution of findings in the absence of a contiguous focus of skin disruption would strongly support neuroarthropathy. Parameters that have been successfully correlated with acute infection superimposed on neuroarthropathy include diffuse bone marrow signal abnormality, progressive subarticular enhancement, loss of subchondral cysts, and the presence of the MRI "ghost sign." PMID- 21644200 TI - Musculoskeletal manifestations of chronic anemias. AB - This article provides an overview of the current use of diagnostic imaging modalities in the evaluation of a heterogeneous group of disorders causing chronic anemias by impaired blood cell production (inherited bone marrow failure syndromes of childhood, aplastic anemia and myelodysplastic syndromes, beta thalassemia) or increased blood cell destruction (sickle cell disease). During the course of these disorders, various musculoskeletal abnormalities can be encountered, including marrow hyperplasia, reversion of yellow marrow to red marrow, growth disturbances, and, occasionally, extramedullary hematopoiesis. Diagnostic imaging may help the clinician to identify specific complications related to either the disease (e.g., bone infarction and acute osteomyelitis in sickle cell disease) or transfusion (e.g., iron overload due to increased hemolysis) and iron chelation (e.g., desferrioxamine-related dysplastic bone changes and deferiprone-related degenerative arthritis) treatments. In this field, magnetic resonance imaging plays a pivotal role because of its high tissue contrast that enables early assessment of bone marrow changes before they become apparent on plain films or computed tomography or metabolic changes occur on bone scintigraphy or positron emission tomography scan. Overall, familiarity with the range of radiological appearances in chronic anemias is important to diagnose complications and establish appropriate therapy. PMID- 21644201 TI - The role of MR imaging in avascular necrosis of the femoral head. AB - Due to the pattern of its blood supply, the femoral head is particularly vulnerable to avascular necrosis (AVN). Nontraumatic AVN is a devastating disorder affecting young patients, and despite treatment it normally follows a progressive course toward a destructive osteoarthropathy. Magnetic resonance (MR) imaging is currently used in major classification systems solely for early detection of femoral head AVN when plain radiographs are normal. More recent data have shown that MR imaging may improve staging, investigate radiologically occult collapse, depict other causes of disability and pain, assess prognosis, and evaluate treatment. This article reviews the established and evolving role of MR imaging in patients at risk or with known femoral head AVN. PMID- 21644202 TI - From Gaucher's disease to metabolic radiology: translational radiological research and clinical practice. AB - Imaging has an increasing role in the management of patients with inborn errors of metabolism. This role is related to expensive enzyme replacement therapy that requires a surrogate biomarker, such as magnetic resonance imaging. This review paper raises the issue of the potential for metabolic radiology to become a subspecialty. PMID- 21644203 TI - Genetic diversity and mycelial compatibility groups of the plant-pathogenic fungus Sclerotinia sclerotiorum in Brazil. AB - The genetic variability of 40 Sclerotinia sclerotiorum isolates from various fields widely distributed throughout Brazil and different host crops was analyzed using RAPD markers and mycelial compatibility groupings (MCGs). The isolates were characterized using 16 random primers of the OPERON series, which produced 121 DNA fragments. UPGMA cluster analysis using Jaccard's genetic distance and MCGs allowed separation of the isolates into three clusters, with similarity indices of 68.2, 61.8, and 61.8%, and five MCGs. The haplotypes obtained with RAPD markers provided very characteristic groupings of S. sclerotiorum isolates according to MCG, but did not show any relationship with geographic origin or host type. Furthermore, analysis of molecular variance demonstrated that 99.1% of the observed variation was a result of genetic differences between individuals; the host culture did not have a significant effect. This is the first report of high level variability of S. sclerotiorum in Brazil based on the study of isolates of wide geographical origin, supported by RAPD markers and MCGs. These results endorse the prevalence of sexual reproduction in tropical and subtropical regions in contrast to clonal reproduction in temperate regions. PMID- 21644204 TI - Association between ACP(1) genetic polymorphism and favism. AB - An association between favism (a hemolytic reaction to consumption of fava beans), glucose-6-phosphate dehydrogenase deficiency (G6PD(-)) and acid phosphatase locus 1 (ACP(1)) phenotypes has been reported; the frequency of carriers of the p(a) and p(c) ACP(1) alleles was found to be significantly higher in G6PD(-) individuals showing favism than in the general population. Here, we investigated the hypothesis that favism is caused by toxic Vicia faba substances, which in some ACP(1) phenotypes cause increased phosphorylation and consequently increased glycolysis, with strong reduction in reduced glutathione production, resulting in hemolysis. It has been demonstrated that ACP(1) f isoforms have physiological functions different from those of s isoforms and are responsible for most of the phosphatase activity, in addition to being less stable in the presence of oxidizing molecules. Thus, the C, CA and A phenotypes, characterized by lower concentrations of f isoforms, could be more susceptible to damage by oxidative events compared to the other phenotypes. To test this hypothesis, the (f+s) enzymatic activity of different ACP(1) phenotypes with and without added V. faba extract was analyzed. Enzymatic activities of ACP(1) A, -CA, -C groups (low activity) and -B, -BA, -CB groups (high activity) were significantly different after addition of V. faba extract. Phenotypes A, CA and C had extremely low enzymatic activity levels, which would lead to low levels of reduced glutathione and bring about erythrocyte lysis. PMID- 21644205 TI - Characterization of 10 novel microsatellite loci for the brown marbled grouper, Epinephelus fuscoguttatus (Serranidae). AB - Epinephelus fuscoguttatus is a commercially important marine fish species in southeast Asia. Due to overfishing and water pollution, this species has been declared as near-threatened. Thus, to provide information to help maintain and preserve the species, microsatellites were developed, using an enriched genomic library method. Thirty individuals were collected from the hatchery of the Fishery Research Institute, Terengganu, Malaysia. These individuals, from four to six years old, originated from Sabah and are maintained in captive culture as broodstock. Genomic DNA was extracted from the fins of selected individuals that weighed 3-8 kg. Ten microsatellite loci were found to be polymorphic in this population, with 5 to 21 alleles per locus. Observed and expected heterozygosities ranged from 0.53 to 0.97 and 0.59 to 0.95, respectively. Only one locus deviated significantly from Hardy-Weinberg equilibrium and no significant linkage disequilibrium was found among the pairs of loci. These polymorphic microsatellite loci will be used by the Malaysian Fishery Research Institute for investigating genetic diversity and for developing breeding strategies. PMID- 21644206 TI - Molecular mapping of quantitative trait loci for drought tolerance in maize plants. AB - Drought tolerance is one of the most important but complex traits of crops. We looked for quantitative trait loci (QTLs) that affect drought tolerance in maize. Two maize inbreds and their advanced lines were evaluated for drought-related traits. A genetic linkage map developed using RFLP markers was used to identify QTLs associated with drought-related traits. Twenty-two QTLs were detected, with a minimum of one and a maximum of nine for drought-related traits. A single-QTL was detected for sugar concentration accounting for about 52.2% of the phenotypic variation on chromosome 6. A single-QTL was also identified for each of the traits root density, root dry weight, total biomass, relative water content, and leaf abscisic acid content, on chromosomes 1 and 7, contributing to 24, 0.2, 0.4, 7, and 19% of the phenotypic variance, respectively. Three QTLs were identified for grain yield on chromosomes 1, 5, and 9, explaining 75% of the observed phenotypic variability, whereas four QTLs were detected for osmotic potential on chromosomes 1, 3, and 9, together accounting for 50% of the phenotypic variance. Nine QTLs were detected for leaf surface area on chromosomes 3 and 9, with various degrees of phenotypic variance, ranging from 25.8 to 42.2%. Four major clusters of QTLs were identified on chromosomes 1, 3, 7, and 9. A QTL for yield on chromosome 1 was found co-locating with the QTLs for root traits, total biomass, and osmotic potential in a region of about 15 cM. A cluster of QTLs for leaf surface area were coincident with a QTL for osmotic potential on chromosome 3. The QTLs for leaf area also clustered on chromosome 9, whereas QTLs for leaf abscisic acid content and relative water content coincided on chromosome 7, 10 cM apart. Co-location of QTLs for different traits indicates potential pleiotropism or tight linkage, which may be useful for indirect selection in maize improvement for drought tolerance. PMID- 21644207 TI - Association of TSC gene variants and hypertension in Mongolian and Han populations. AB - We investigated a possible association between genetic variations in the thiazide sensitive Na-Cl cotransporter (TSC) gene and essential hypertension (EH) in the Mongolian and Han ethnic groups in Inner Mongolia. Our study included 385 unrelated Mongolian herdsmen and 523 Han farmers. Nine tagSNPs of TSC were identified from the Chinese HapMap database based on pairwise r(2) >= 0.5 and minor allele frequency >=0.05. Genotyping was performed using the PCR/ligase detection reaction assay. Association between tagSNPs and hypertension was investigated under the additive model. There were significant differences between the genotype and allele frequencies of rs13306673 between the EH group and the control group in the Han population. Significant associations were found between the rs7204044 variant and EH in both the Mongolian and Han ethnic groups. The frequency of haplotype GCA in the EH group was significantly higher than in the control group in the Mongolian population. In the Han population, the frequency of haplotype TGG was significantly higher in the EH group than in controls, whereas haplotype TGA occurred significantly less often in EH than in controls. We suggest that rs7204044 of TSC is a genetic factor for EH in these two ethnicities and that rs13306673 is a genetic factor for EH in the Han population. PMID- 21644208 TI - Identification of 18 genes encoding necrosis-inducing proteins from the plant pathogen Phytophthora capsici (Pythiaceae: Oomycetes). AB - Phytophthora capsici is an aggressive plant pathogen that affects solanaceous and cucurbitaceous hosts. Necrosis-inducing Phytophthora proteins (NPPs) are a group of secreted toxins found particularly in oomycetes. Several NPPs from Phytophthora species trigger plant cell death and activate host defense gene expression. We isolated 18 P. capsici NPP genes, of which 12 were active during hypha growth from a Phytophthora stain isolated from pepper (Capsicum annuum) plants in China. The 18 predicted proteins had a sequence homology of 46.26%. The 18 Pcnpp sequences had a conserved GHRHDWE motif and fell into two groups. Eleven sequences in group 1 had two conserved cysteine residues, whereas the other seven sequences in group 2 lacked these two cysteine residues. A phylogenetic tree was constructed on the basis of the alignment of the predicted protein sequences of 52 selected NPP genes from oomycetes, fungi and bacteria from Genbank. The tree did not rigorously follow the taxonomic classification of the species; all the NPPs from oomycetes formed their own clusters, while fungal sequences were grouped into two separate clades, indicating that based on NPPs, we can separate oomycetes from fungi and bacteria, and that expansion of the NPP family was a feature of Phytophthora evolution. PMID- 21644209 TI - Genetic assessment of the Atlantic Forest bristle porcupine, Chaetomys subspinosus (Rodentia: Erethizontidae), an endemic species threatened with extinction. AB - The bristle-spined porcupine, Chaetomys subspinosus, an endemic rodent from Atlantic Forest, was considered to be abundant in the recent past, but population reductions due to habitat loss and expansion of human activities caused this species to be included in the "vulnerable" category of the World Conservation Union Red List. We performed the first genetic assessment in natural populations of this focal species along its geographical distribution. Thirty-five non invasive samples (hair) were collected from three natural populations in the Brazilian States of Sergipe, Bahia and Espirito Santo. Genetic similarity obtained by Jaccard's index, based on dominant RAPD and ISSR markers, varied between 25 and 100%. Four clusters, mainly coincident with the geographical distribution of the populations, were observed. Analysis of molecular variance based on 47 polymorphic loci showed that there was 15.99% genetic variability among populations and 84.01% within populations. The estimated genetic structure among populations (Phi(ST)) was 0.16. The populations may have formed a continuum along the past distribution of the Atlantic rainforest but historical events of human occupation resulted in recent divergence among sampled populations. PMID- 21644210 TI - Development of a novel and efficient strategy for practical identification of Pyrus spp (Rosaceae) cultivars using RAPD fingerprints. AB - Accurate and reliable cultivar identification of crop species is essential to guarantee plant material identity for purposes of registration, cultivar protection and production. To facilitate identification of plant cultivars, we developed a novel strategy for efficient recording of DNA molecular fingerprints in genotyped plant individuals. These fingerprints can be used as efficient referential information for quick plant identification. We made a random amplified polymorphic DNA (RAPD) marker analysis of 68 pear cultivars. All pear genotypes could be distinguished by a combination of eight 11-mer primers. The efficiency of the method was further verified by correct identification of four cultivars randomly chosen from the initial 68. The advantages of this identification include use of fewer primers and ease of cultivar separation by the corresponding primers marked on the cultivar identification diagram. The cultivar identification diagram can efficiently serve for pear cultivar identification by readily providing the information needed to separate cultivars. To the best of our knowledge, this is the most efficient strategy for identification of plant varieties using DNA markers; it could be employed for the development of the pear industry and for the utilization of DNA markers to identify other plant species. PMID- 21644211 TI - Association of variable number of tandem repeats polymorphism in the IL-4 gene with end-stage renal disease in Malaysian patients. AB - Variable number of tandem repeats (VNTR) polymorphism in the interleukin 4 (IL-4) gene has been associated with end-stage renal disease (ESRD) subjects in many different populations, although with conflicting results. We determined the 70 bp of VNTR polymorphism at intron 3 of the IL-4 gene in Malaysian ESRD subjects. Buccal cells were collected from 160 case and 160 control subjects; genomic DNA was amplified using PCR, followed by agarose gel electrophoresis. There were significant differences in genotypes and alleles of the IL-4 gene. We conclude that VNTR polymorphism of the IL-4 gene is a risk factor for the development of ESRD among Malaysians. PMID- 21644212 TI - Lack of association of variants of the renal salt reabsorption-related genes SLC12A3 and ClC-Kb and hypertension in Mongolian and Han populations in Inner Mongolia. AB - Abnormalities in renal sodium chloride and water reabsorption play important roles in the development of hypertension. Mutations in the genes involved in renal sodium chloride reabsorption can affect blood pressure. Recently, the R904Q variant of the sodium/chloride transporters, member 3 (SLC12A3) gene and the T481S variant of the chloride channel Kb (ClC-Kb) gene were found to be implicated in essential hypertension. We investigated a possible role of the SLC12A3 and ClC-Kb genes in the prevalence of essential hypertension in the Mongolian and Han ethnic groups. The study population comprised 308 unrelated Mongolians with essential hypertension, 271 Mongolian normotensives, 285 unrelated Han with essential hypertension, and 194 Han normotensives living in Inner Mongolia. The presence of the SLC12A3 R904Q and ClC-Kb-T481S polymorphisms was determined using TaqMan PCR. The risk factors for hypertension were age, body mass index, alcohol consumption, total plasma cholesterol, and low-density lipoprotein cholesterol. The genotype and allele frequencies of SLC12A3 R904Q and ClC-Kb-T481S were not significantly different between hypertensive patients and controls in the Mongolian (SLC12A3 R904Q, P = 0.471 and P = 0.494, ClC-Kb-T481S, P = 0.960 and P = 0.960, respectively) and Han (SLC12A3 R904Q, P = 0.765 and P = 0.777, ClC-Kb-T481S, P = 0.100 and P = 0.103, respectively) populations. There was no significant association between the SLC12A3 R904Q variant and the ClC-Kb T481S variant and essential hypertension in either ethnic group. PMID- 21644213 TI - Identification of novel and recurrent mutations in the calcium binding type III repeats of cartilage oligomeric matrix protein in patients with pseudoachondroplasia. AB - Pseudoachondroplasia is an autosomal dominant osteochondrodysplasia characterized by disproportionate short stature, joint laxity, and early onset osteoarthrosis. Pseudoachondroplasia is caused by mutations in the gene encoding cartilage oligomeric matrix protein (COMP). We looked for mutations in the COMP gene in three sporadic Chinese pseudoachondroplasia patients and identified two novel mutations, c.1189G>T (p.D397Y) and c.1220G>A (p.C407Y), and one recurrent mutation, c.1318G>C (p.G440R), in the calcium binding type III repeats of COMP. This study confirms the relationship between mutations of the COMP gene and clinical findings of pseudoachondroplasia; it also provides evidence for the importance of the calcium binding domains to the functioning of COMP. PMID- 21644214 TI - Genetic analysis of 30 InDel markers for forensic use in five different Chinese populations. AB - Allele frequencies of 30 insertion/deletion polymorphism (InDel) markers previously selected and validated for forensic purposes were assessed in 419 unrelated individuals originating from five different populations of P.R. China, including Chinese Han, Chinese Hui, Uighur, Mongolians, and Tibetans. Hardy Weinberg equilibrium tests and linkage disequilibrium analysis were performed; the allele frequency distributions of the 30 InDel markers met the conditions for genetic equilibrium in all five populations and the InDel markers on the same chromosome did not generate any linkage blocking. Analysis of molecular variance indicated that genetic variation among the five populations represents only 4% of the total genetic diversity. We determined the cumulative power of discrimination for each population: 0.99999999999841 in Chinese Han, 0.99999999999690 in Chinese Hui, 0.99999999999709 in Uighur, 0.99999999999772 in Mongolians, and 0.99999999999854 in Tibetans. PMID- 21644215 TI - [Identification of a novel mutation of GALNS gene from a Chinese pedigree with mucopolysaccharidosis type IV A]. AB - OBJECTIVE: To study the molecular genetic mechanism of mucopolysaccharidosis type IV A(MPS IV A), and reveal the relationship between the genotype and phenotype, and provide a basis for prenatal gene diagnosis in the future. METHODS: A preliminary diagnosis was made by qualitative detection of urinary glycosaminoglycans of the suspected MPS IV A proband. Then, mutation detection was performed on the proband and her family members with PCR and direct sequencing of the PCR products. After a novel c.1567T to G mutation was detected, Xsp I restriction enzyme digestion and amplification refractory mutation system (ARMS) fast specific identification were established to analyze the sequences of exon 14 in GALNS gene, including 110 randomly selected healthy controls, the proband and other pedigree members. At the same time, bioinformatic approaches for protein secondary, tertiary structure prediction were applied to identify the novel pathologic mutation. RESULTS: The proband's urine GAGs test was a weak positive(+/- ), and a c.1567T to G heterozygous termination codon mutation in exon 14 and a c.374C to T heterozygous missense mutation in exon 4 were found. The proband was compound heterozygous of the two mutations, so was her younger sister. Her mother was a carrier with only a c.1567T to G heterozygous mutation in exon 14. Her father had a heterozygous mutation of c.374C to T in exon 4. After Xsp I restriction enzyme digestion, healthy controls had three bands including 28 bp, 120 bp and 399 bp, while the proband and her mother had four bands consisting of 28 bp, 120 bp, 148 bp and 399 bp. For amplification by ARMS specific primers, it was negative for the controls, while it was positive for the proband and the carrier. The results of protein secondary and tertiary structure prediction showed that the c.1567T to G mutation located in the stop codon, resulted in stop codon (TAG) changing to glutamic acid (GAG), with the peptide chain extending 92 amino acid residues, and secondary and tertiary protein structure change, which were not found in the controls. The result of enzyme assay showed that the activity of GALNS enzyme in the affected child was 8.3 nmol/17h/mg pr, which was obviously lower than the normal value (the normal range is 41.9-92.1 nmol/17h/mg pr). CONCLUSION: These results illustrate that the c.1567 T to G is a novel pathologic mutation, which is the main cause of the disease in this family. PMID- 21644216 TI - [Two cases of partial trisomy 8p derived from paternal reciprocal translocation or maternal insertion translocation: clinical features and genetic abnormalities]. AB - OBJECTIVE: To determine the origin of aberrant chromosomes involving the short arm of chromosome 8 in two mentally retarded children, and to correlate the karyotype with abnormal phenotype. METHODS: Routine G-banding was performed to analyze the karyotypes of the two patients and their parents, and array comparative genomic hybridization (array CGH) was used for the first patient for fine mapping of the aberrant region. RESULTS: The first patient presented with only mental retardation. The father had normal karyotype. The mother had an apparent insertion translocation involving chromosomes 8 and 3 [46, XX, inv ins (3; 8) (q25.3; p23.1p11.2)], the karyotype of the child was ascertained as 46, XX, der(3) inv ins (3; 8)(q25.3; p23.1p11.2). Array CGH finely mapped the duplication to 8p11.21-8p22, a 26.9 Mb region. The other patient presented with mental retardation, craniofacial defects, congenital heart disease and minor skeletal abnormality. The mother had normal karyotype. The father had an apparently balanced translocation involving chromosome 8p and 11q, the karyotype was 46, XY, t(8; 11)(p11.2; q25). The karyotype of the child was then ascertained as 46, XX, der(11)t(8; 11)(p11.2; q25). CONCLUSION: These results suggested that partial trisomy 8p was primary cause for the phenotypic abnormalities of the two patients, whereas a mild phenotypic effect was observed in patient 1. Parental karyotype analysis could help define the aberrant type and recurrent risk evaluation. In contract to routine karyotype analysis, aberrant regions could be mapped by array CGH with higher resolution and accuracy. PMID- 21644217 TI - [Mutation screening and prenatal diagnosis of a pedigree with Glanzmann's thrombasthenia]. AB - OBJECTIVE: Mutation screening was performed in a pedigree of Glanzmann's thrombasthenia (GT) and prenatal diagnosis was performed. METHODS: In this study, reverse transcription-PCR-sequencing and PCR-sequencing, as well as restriction fragment length polymorphism(RFLP) and A/T-cloned-sequencing, were used to screen the ITGA2B and ITGB3 mutation in a pedigree with Glanzmann's thrombasthenia in the RNA and DNA level. Prenatal diagnosis was performed for this pedigree. RESULTS: Deletion of 99 bps was found in the cDNA of the patient in the pedigree, leading to deletion of 33 codons (from codon 160 to 192). After genomic analysis, the patient was found to be a compound heterozygote of c.374C to G mutation and intron 4(IVS-4) + 5 G to C mutation. The two mutations were inherited from the parents. IVS-4 + 5 G to C mutation was a point mutation in the splice site, while c.374C to G mutation was out of the splice site. But both of them resulted in the same splice pattern in RNA. The two mutations were novel mutations which have not been reported in Human Gene Mutation Database (HGMD) and the mutation data base of Glanzmann's thrombasthenia. The results of ITGB3 gene screening is normal in the proband and his parents. CONCLUSION: Two novel mutation, c.374C to G and IVS 4 + 5 G to C were found in this study, which might be the cause of GT in the pedigree. PMID- 21644218 TI - [Linkage analysis of susceptibility loci in 2 target chromosomes in pedigrees with paranoid schizophrenia and undifferentiated schizophrenia]. AB - OBJECTIVE: To investigate the relationship of susceptibility loci in chromosomes 1q21-25 and 6p21-25 and schizophrenia subtypes in Chinese population. METHODS: A genomic scan and parametric and non-parametric analyses were performed on 242 individuals from 36 schizophrenia pedigrees, including 19 paranoid schizophrenia and 17 undifferentiated schizophrenia pedigrees, from Henan province of China using 5 microsatellite markers in the chromosome region 1q21-25 and 8 microsatellite markers in the chromosome region 6p21-25, which were the candidates of previous studies. All affected subjects were diagnosed and typed according to the criteria of the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revised (DSM-IV-TR; American Psychiatric Association, 2000). All subjects signed informed consent. RESULTS: In chromosome 1, parametric analysis under the dominant inheritance mode of all 36 pedigrees showed that the maximum multi-point heterogeneity Log of odds score method (HLOD) score was 1.33 (alpha = 0.38). The non-parametric analysis and the single point and multi-point nonparametric linkage (NPL) scores suggested linkage at D1S484, D1S2878, and D1S196. In the 19 paranoid schizophrenias pedigrees, linkage was not observed for any of the 5 markers. In the 17 undifferentiated schizophrenia pedigrees, the multi-point NPL score was 1.60 (P= 0.0367) at D1S484. The single point NPL score was 1.95(P= 0.0145) and the multi-point NPL score was 2.39 (P= 0.0041) at D1S2878. Additionally, the multi-point NPL score was 1.74 (P= 0.0255) at D1S196. These same three loci showed suggestive linkage during the integrative analysis of all 36 pedigrees. In chromosome 6, parametric linkage analysis under the dominant and recessive inheritance and the non-parametric linkage analysis of all 36 pedigrees and the 17 undifferentiated schizophrenia pedigrees, linkage was not observed for any of the 8 markers. In the 19 paranoid schizophrenias pedigrees, parametric analysis showed that under recessive inheritance mode the maximum single-point HLOD score was 1.26 (alpha = 0.40) and the multi-point HLOD was 1.12 (alpha = 0.38) at D6S289 in the chromosome 6p23. In nonparametric analysis, the single-point NPL score was 1.52 (P= 0.0402) and the multi-point NPL score was 1.92 (P= 0.0206) at D6S289. CONCLUSION: Susceptibility genes correlated with undifferentiated schizophrenia pedigrees from D1S484, D1S2878, D1S196 loci, and those correlated with paranoid schizophrenia pedigrees from D6S289 locus are likely present in chromosome regions 1q23.3 and 1q24.2, and chromosome region 6p23, respectively. PMID- 21644219 TI - [Application of enzyme assay and gene analysis in the prenatal diagnosis for a family with glycogen storage disease type II]. AB - OBJECTIVE: To carry out prenatal diagnosis for a glycogen storage disease type II (GSD II ) affected family. METHODS: The acid-alpha -glucosidase (GAA) activity was measured in whole leukocytes and cultured amniocytes with 4 methylumbelliferyl-alpha -D-glucopyranoside as substrate and with acarbose as inhibitor. The coding regions of GAA gene were amplified by polymerase chain reaction and analyzed by direct DNA sequencing. RESULTS: The proband and the fetus had low GAA activity (12.3% and 1.1% of the average normal range, respectively). Mutation analysis of the GAA gene revealed a novel nonsense mutation p.W738X and a reported nonsense mutation p.E888X in both the proband and the fetus; the reported pseudodeficiency allele c.[1726G to A: 2065G to A] was found in the proband, the mother and the fetus. CONCLUSION: The proband and the fetus were both GSD II affected. A combination of GAA activity analysis and mutation analysis is efficient for the prenatal diagnosis of GSD II. Mutation analysis should be a routine method in the prenatal diagnosis of GSD II in Asian population, where pseudodeficiency allele can cause low GAA activity in normal individuals which is relatively common in Asian. PMID- 21644220 TI - [The association between paternal age and schizophrenia in a Chinese Han population]. AB - OBJECTIVE: To investigate whether advanced paternal age is related to an increased risk of schizophrenia in Chinese Han population. METHODS: A case control design study was performed. Three hundred and fifty-one patients with schizophrenia and 199 unrelated healthy volunteers were recruited. By using Logistic regression, paternal age was divided into five categories, and maternal age into four categories. Setting the paternal age of 26-30 years as reference, the OR, P values and 95% CI of the other paternal age categories were analyzed, respectively. The participant's sex, age and parental age at birth were used as covariants for adjusting confounding effects. RESULTS: The OR for schizophrenia in offspring whose paternal age at birth of 31-35 years, 36-40 years, and >= 41 years categories were 3.834, 8.805, and 11.619 respectively. The advanced maternal age had no significant effects on the risk for schizophrenia in offspring. CONCLUSION: The advanced paternal age was associated with elevated risk for schizophrenia in offspring among a Han Chinese population. Putative biological mechanisms may include accumulated de novo mutations and alterations in epigenetic regulations with aging in spermatogenesis. PMID- 21644221 TI - [Association between single nucleotide polymorphisms of 5'-untranslated region of GPx4 gene and male infertility]. AB - OBJECTIVE: To study the association between the single nucleotide polymorphisms (SNPs) of the 5'-untranslated region (5'-UTR) of phospholipid hydroperoxide glutathione peroxidase (GPx4 or PHGPx) gene and oligo- or asthenozoospermic male infertility. METHODS: The 5'-UTR region of the GPx4 gene was amplified from infertile men and controls using the polymerase chain reaction and was analyzed for polymorphisms by direct sequencing. RESULTS: A total of 9 SNPs were present in the cohort, however there were no significant differences in these 9 SNPs between the case and control groups. According to the results of linkage disequilibrium analysis and haplotype construction, one haplotype (rs757229 rs757230-rs4588110-rs3746165-rs3746166: C-G-G-T-A) was present only in the control men, and significant difference was detected(P< 0.01). CONCLUSION: The SNPs of 5'-UTR region of the GPx4 gene might not be associated with oligo- or asthenozoospermic male infertility. However, the haplotype (rs757229-rs757230 rs4588110- rs3746165-rs3746166: C-G-G-T-A) might be a protective haplotype. PMID- 21644222 TI - [Advance on Dicer gene and its role in female reproduction]. AB - Dicer is an RNAse III endonuclease that is essential for the biogenesis of microRNAs and small interfering RNAs. These small RNAs transcriptionally and post transcriptionally regulate mRNA expression through RNA interference mechanisms. Recently, the function of Dicer in female reproduction has begun to be elucidated through the use of knockout mouse models. Several latest studies have indicated that Dicer gene plays a key role in female reproductive processes such as oocyte maturation, early embryonic development and implantation and steroidgenesis. When Dicer expression is decreased in female reproductive tissues or cells, it will cause infertility. In this article, author discuss the role of Dicer gene in female reproductive tract, and advance of Dicer gene study in female reproductive events. PMID- 21644223 TI - [Association of genetic polymorphisms in methionine metabolism genes with X linked adrenoleukodystrophy]. AB - OBJECTIVE: To investigate the association of the polymorphisms of methionine metabolism genes and the phenotype of X-linked adrenoleukodystrophy (X-ALD) and clinical severity. METHODS: The clinical information of 120 X-ALD patients were analyzed and three genetic variants involved in the methionine metabolism, including cystathionine beta-synthase (CBS) c.844_855ins68, 5 methyltetrahydrofolate-homocysteine-S-methyltransferase (MTR) c.2756A to G, and transcobalamin 2 (TC2) c.776 C to G were analyzed by polymerase chain reaction and sequencing. The association between these polymorphisms and phenotype of X ALD was studied. RESULTS: The frequency of GG genotype of the TC2 c.776 C/G was higher in patients with central nervous system(CNS) demyelination than in controls (P= 0.012). However, the other two polymorphisms did not show any significant associations with the phenotypes. CONCLUSION: The GG genotype of TC2 c.776 C/G may contribute to X-ALD phenotype. PMID- 21644224 TI - [Cytogenetic study of mantle cell lymphoma]. AB - OBJECTIVE: To explore the cytogenetic features of mantle cell lymphoma (MCL). METHODS: Bone marrow cells from 18 MCL patients with bone marrow invasion were cultured for 24 hours, then routine karyotype analysis was performed with R banding technique. Interphase fluorescence in situ hybridization (FISH) and a panel of 5 probes, including CCND1/IgH, CEP12, D13S319, p53 gene and ATM gene, were used to investigate the cytogenetic features of the samples. RESULTS: Chromosome aberrations were found in 9 (64.3%, 9/14) patients by conventional cytogenetics (CC), 8(57.1%, 8/14) patients had the aberration of t(11; 14), 6(42.9%, 6/14) had complex aberrant karyotypes, of which 2 (14.3%, 2/14) had highly complex aberrant karyotypes. A total of 28 abnormalities were detected, among them 19 (67.9%) were structural abnormalities, the other 9 (32.1%) were numerical aberrations. The aberration of t(11; 14) was found in all 18 (100%) patients with MCL by FISH. Secondary cytogenetic aberrations were detected in 14 patients by FISH. The most common abnormality was del(11q22.3) (57.1%), the rate of aberrations for del(17p13), + 12 and del(13q14) were 42.9%, 35.7% and 21.4%, respectively. Two (14.3%) and 4 (28.6%) patients were detected to have combinations of 2 and 3 aberrations. CONCLUSION: In addition to t(11; 14), most MCL patients have other chromosome aberrations, especially complex aberrant karyotype. PMID- 21644225 TI - [Association of the genetic variations of bone morphogenetic protein 7 gene with diabetes and insulin resistance in Xinjiang Uygur population]. AB - OBJECTIVE: To investigate the association between the genetic variations of the functional region in bone morphogenetic protein gene (BMP7) with type 2 diabetes mellitus in Chinese Uygur individuals. METHODS: A case-control study was conducted based on epidemiological investigation. A total of 717 Uygur subjects (276 males and 441 females) were selected and divided into two groups: diabetes mellitus group (n = 502, 191 males and 311 females) and control group (n = 215, 85 males and 130 females). All exons, flanking introns and the promoter regions of (BMP7) gene were sequenced in 48 Uygur diabetics. Representative variations were selected according to the minor allele frequency (MAF) and linkage disequilibrium and genotyped using the TaqMan polymerase chain reaction method in 717 Uygur individuals, a relatively isolated general population in a relatively homogeneous environment and a case-control study was conducted to test the association between the genetic variations of (BMP7) gene and type 2 diabetes mellitus. RESULTS: Five novel and 8 known variations in the (BMP7) gene were identified. All genotype distributions were tested for deviations from Hardy Weinberg equilibrium (P> 0.05). There was significant difference of genotype distribution of rs6025422 between type 2 diabetes mellitus and control groups in the male population (P< 0.05, P adjusted > 0.05), but there was no difference in total and female population (P> 0.05). And the means of fasting blood glucose (FBG), fasting insulin and HOMA-index significantly decreased in individuals with AA, AG and GG genotypes of rs6025422 in male population (P< 0.05), but not in total and female population (P> 0.05). The logistic regression analysis showed that GG genotype of rs6025422 variation might be a protective factor for diabetes in male (OR= 0.637, 95% confidence interval 0.439-0.923, P< 0.05). CONCLUSION: The present study suggests that the rs6025422 polymorphism in (BMP7) gene may be associated with diabetes mellitus and insulin resistance in Uygur men. PMID- 21644226 TI - [Identification of a novel allele HLA-B*15:129 by polymerase chain reaction with allele group-specific primers]. AB - OBJECTIVE: To analyze the sequence of the exons 2-4 of human leukocyte antigen (HLA) novel allele HLA-B*15:129. METHODS: DNA of the proband was extracted from whole blood by commercial DNA extraction kit. The amplification for HLA-B exons 2 4 was performed separately by polymerase chain reaction (PCR) with allele group specific primers. The PCR products were digested with enzymes and then directly sequenced for exons 2-4 of HLA-B locus in both directions. RESULTS: Sequencing results showed the HLA-B alleles of the proband included B*07:02 and a novel allele. The sequence of the novel allele has been submitted to GenBank (accession no. EF473219) and the allele has been officially named B*15:129 by the WHO Nomenclature Committee. Comparing with the HLA-B*15:01:01:01, the sequence of exons 2-4 of HLA-B*15:129 showed three nucleotide difference in exon 3 at positions 362 and 363 from GG to AT and positions 369 from C to T, which resulted in an amino acid change from Arg to Asn at codon 97. CONCLUSION: A novel HLA-B allele was identified and has been officially named B15:129 by the WHO Nomenclature Committee. PMID- 21644227 TI - [Association of HLA DRB1 polymorphism with susceptibility to myelodysplastic syndrome and aplastic anemia in Chinese Han population]. AB - OBJECTIVE: To investigate the association of the HLA-DRB1 polymorphism with susceptibility to myelodysplastic syndrome (MDS) and aplastic anemia (AA) in Chinese Han population. METHODS: The polymorphism of HLA-DRB1 alleles in 242 patients with MDS, 115 patients with AA and 2264 umbilical cord blood control samples were tested by polymerase chain reaction-sequence specific primer (PCR SSP). RESULTS: Compared with normal controls, the frequency of HLA-DRB1*15 was significantly increased in the MDS group and AA group (22.93% vs. 17.25%, Chi square = 9.662, OR= 1.428, P= 0.003; 26.52% vs. 17.25%, Chi-square = 12.924, OR= 1.732, P= 0.001). And this is mainly due to the increase in the male patients in both patient groups: in the MDS males, 24.68% vs. 17.25%, Chi-square= 11.194, OR= 1.572, P= 0.001, and in the AA males, 29.29% vs. 17.25%, Chi-square= 13.563, OR= 1.987, P= 0.001. No significant difference between the controls and the female patients in both groups was observed. CONCLUSION: The HLA-DRB1*15 could be a susceptibility allele for MDS and AA male patients. PMID- 21644228 TI - [Cloning and sequencing analysis of a novel allele HLA-A*02:251]. AB - OBJECTIVE: To identify a novel human leukocyte antigen (HLA) allele A*02:251 and analyze the sequences in Chinese population. METHODS: Routine HLA-A, -B, -DRB1 high resolution genotyping for healthy Chinese donors and patients was performed with polymerase chain reaction-sequence based typing. An unknown HLA-A allele was initially detected by HLA typing in the healthy donor. Genomic DNA of the HLA-A locus in the proband was amplified, the amplified product was cloned by PMD18-T to split the two alleles, and selected clones were sequenced. RESULTS: The sequencing results showed that a normal A*02:06:01 and a novel A*02:251 variant allele were identified. The sequence of the novel allele has been submitted to GenBank (HM245348). Nucleotide sequence alignments with HLA-A allele from the IMGT/HLA Sequence Database showed that the novel A*02 variant allele differed from the closest allele A*02:01:01:01 by nt 383 G to C (codon 128 GAG to GAC) in exon 3, which resulted in one amino acid substitution of Glu to Asp. The HLA-A, B, C and DQB1 alleles of the healthy donor did not match with that of the patient. CONCLUSION: This novel allele is officially designated as HLA-A*02:251 by World Health Organization(WHO) Nomenclature Committee (Submission ID HWS10010755). The sequence of HLA-A locus in exon 3 is confirmed to be polymorphic in Chinese population. PMID- 21644229 TI - [Association of single nucleotide polymorphism in CYP17 and ERalpha genes with endometriosis risk in southern Chinese women]. AB - OBJECTIVE: To investigate the association of single nucleotide polymorphisms in cytochrome P450 17 (CYP17) and estrogen receptor alpha (ERalpha ) genes with the risk of endometriosis among southern Chinese women. METHODS: Two SNPs rs743572 (CYP17 gene 34T/C) and rs9322331 (ERalpha gene -397T/C) were genotyped by high resolution melting curve in 432 endometriosis patients and 499 matched controls. RESULTS: There was no significant difference in the genotype frequencies of the two loci between endometriosis patients and the control subjects (P> 0.05). And there was no significant interaction effect of these two genes on the disease either. CONCLUSION: CYP17 gene and ERalpha gene may not be genetic risk factors for endometriosis among southern women in China. PMID- 21644230 TI - [Study of dystrophin gene non-deletion/duplication mutations causing Becker muscular dystrophy]. AB - OBJECTIVE: To identify potential mutations in patients featuring Becker muscular dystrophy (BMD) and to enhance the understanding of non-deletion/duplication mutations of the dystrophin gene causing BMD. METHODS: Clinical data of two patients affected with BMD were collected. Potential mutations in the dystrophin gene were screened with multiplex ligation-dependent probe amplification assay (MLPA). Biopsied muscle samples were examined with HE staining, immnostaining with anti-dystrophin antibody, and electronic microscopy. RESULTS: MLPA assay suggested that both cases were probably due to non-deletion/duplication mutations of the dystrophin gene. Light and electronic microcopy of skeletal muscle biopsies confirmed dystrophic changes in both patients. For patient A, immunostaining showed non-contiguous weak staining for most parts of sarcolemma. For patient B, immunostaining showed positive result with N-terminal anti dystrophin antibody and negative result with C-terminal anti-dystrophin antibody. CONCLUSION: For patients with mild phenotypes but without dystrophin gene deletion/duplication, muscle biopsy and immunochemistry are helpful for diagnosis and prognosis. PMID- 21644231 TI - [Single nucleotide polymorphisms of CTLA4 gene and their association with human cervical cancer]. AB - OBJECTIVE: To evaluate the association between single nucleotide polymorphisms (SNPs) of cytotoxic T-lymphocyte-associated protein 4 (CTLA4) gene and susceptibility to cervical cancer. METHODS: One hundred patients and 100 healthy controls from Hubei province were genotyped for 20 polymorphic loci using Sequenom. RESULTS: The frequency of rs11571316 G allele and rs5742909 T allele, which are localized in the promoter region, and rs11571319 A allele, which is downstream of the gene, were significantly higher in patients than in controls. Luciferase assay showed that, as the previously reported rs5742909 T allele, rs11571316 G allele could significantly increase the expression of the reporter gene. CONCLUSION: SNPs in the promoter region of (CTLA4) gene might increase the susceptibility to cervical cancer by increasing (CTLA4) gene expression. PMID- 21644232 TI - [Association of CASP3 and CASP9 polymorphisms with genetic susceptibility to stomach cancer]. AB - OBJECTIVE: To investigate the association between the apoptosis genes CASP3(rs12108497) and CASP9 (rs4646018) polymorphisms and the risk of developing stomach cancer. METHODS: In this population-based case-controlstudy, 278 cases with stomach cancer and 278 age (+/- 5 years), gender, and residential area matched controls were recruited. The genotypes were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The unconditional Logistic regression analysis was utilized to calculate the odds ratios (OR) and 95% confidence intervals (CI). RESULTS: The individuals with TC, CC genotypes of rs12108497 locus had significantly increased risk of stomach cancer in comparison to those carrying TT genotype (OR= 1.45, 95% CI: 1.01-2.07 for TC; OR= 2.17, 95%CI: 1.15-4.08 for CC). However, the rs4646018 locus of CASP9 gene polymorphism was not related to stomach cancer risk. Compared with the subjects carrying the both low-risk genotypes, those carrying 1 or 2 high-risk genotypes had a noteworthy increased risk of stomach cancer (OR= 1.60, 95% CI: 1.12-2.30). The combined high-risk genotypes appeared to be more evident in subjects of male (OR= 1.62, 95% CI: 1.05-2.49), ever-smokers (OR= 1.87, 95%CI: 1.12-3.12), ever-drinkers (OR= 1.92, 95%CI: 1.02-3.65) and no family history of cancer (OR= 1.78, 95%CI: 1.18-2.68). CONCLUSION: The current findings suggest that the polymorphism of CASP3 rs12108497 might be associated with the risk of stomach cancer. However, the CASP9 rs4646018 polymorphism may not be related to the stomach cancer risk. PMID- 21644233 TI - [Proteomic analysis of human fetal and adult brain cortex]. AB - OBJECTIVE: To study the differences of protein expression levels in the brain cortex of human fetus and adult with proteomics technique, and provide preliminary data on the change of proteins during brain development. METHODS: Proteins extracted from human temporal lobes in fetal (3 month and 5 month respectively) and adult (30 years old) brain were separated by two-dimensional gel electrophoresis (2DE). The proteins were then stained with colloidal Coomassie blue to produce a high-resolution map of the proteiome. The differential protein spots were analyzed by PDQuest 7.0 software and 8 spots, which were gradually reduced or gradually increased in brain development process and the protein spots of difference over two-fold in the brain, were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF/TOF). RESULTS: (1) On average, 642, 511 and 527 protein spots could be obtained in the temporal lobes of adult, 3 month and 5 month fetus. The matching rate of images was 87%. The basic proteins in adult brain were obviously much more than that in the fetus; (2) There were 172, 171 and 152 singular protein spots in temporal lobes of adult, 3 month and 5 month fetus respectively.(3) Compared with adult, there were 131 and 115 different protein spots in the 3 month and 5 month fetus respectively. There were 60 and 40 protein spots with more than 2 fold difference, among which 24 and 17 were down-regulated, and 36 and 23 were up-regulated respectively. (4) There was different expression in proteins such as serum albumin, triosephosphate isomerase, etc. in the 3 groups. Fatty acid binding protein 7 and unnamed proteins were only highly expressed in the 3 month brain; ribulose-1, 5-bisphosphate carboxylase/oxygenase large subunit and transducin beta-1 subunit were up-regulated in adult brain. Serum albumin decreases gradually with brain development. However, ATP synthase, mitochondrial F0 complex, and triosephosphate isomerase increase gradually with brain development. CONCLUSION: The proteins of human brain cortex were obviously changed from embryonic stage to adult. The differentially displayed proteins may provide further insight into the understanding of development of human brain. PMID- 21644234 TI - [Clinical and gene mutation analyses of three patients with ornithine carbamoyltransferase deficiency]. AB - OBJECTIVE: To analyze the clinical and genetic characteristics of three children with ornithine carbamoyltransferase deficiency(OTCD), and to provide a practical method for gene diagnosis and genetic counseling of the disease. METHODS: All exons of the ornithine carbamoyltransferase (OTC) gene were screened by polymerase chain reaction-DNA direct sequencing in the three OTCD patients. RESULTS: One patient firstly presented as vomiting at 6 month of age. A missense mutation of T262I was detected. His mother had the same mutation without any clinical symptoms. The second patient presented as restlessness, and had a missense mutation of R277W. Gene analysis of his parents was not available. The third patient presented as neonatal lethargy, harbored a missense mutation of I172M. His mother had the same mutation without any clinical symptoms. CONCLUSION: Gene mutation analysis is a feasible way for diagnosing OTCD. Patients with I172M mutation present symptom early, while those with T262I and R277W mutations manifest symptoms later. Gene mutation analysis will be important for asymptomatic and prenatal diagnosis and genetic counseling. PMID- 21644235 TI - [Evaluation of Down's syndrome screening methods using maternal serum biochemistry in the second trimester pregnancy]. AB - OBJECTIVE: To provide basis for selecting the suitable method of Down's syndrome biochemical screening in the second trimester pregnancy. METHODS: A total of 30 547 singleton pregnancies between 14 and 20(+ 6) weeks of pregnancy were collected and analyzed for maternal serum alpha-fetoproteins (AFP) and human chorionic gonadotrophin, free beta subunit (beta-HCG) with or without unconjugated estriol (uE3). The screening risks were calculated using the software Lifecycle. The detection rates and the cost of per Down's syndrome detected were calculated and compared. And four different methods were compared in a series of 64 serum samples from Down's syndrome pregnancies. RESULTS: (1) Among the 64 affected cases, the detection rate of Down's syndrome was improved no matter in the double test (DT) or in the triple test (TT) if software Lifecycle (LC) was used to evaluate risks. And it was not suitable to evaluate risks with software 2T-Risks in the triple tests. (2) In the cohort of 30 547 singleton pregnancies, the detection rate of Down's syndrome with project DT-LC, which was double test using AFP and free beta-HCG together with software Lifecycle, and project TT-LC, which was triple test using AFP, free beta-HCG and uE3 together with software Lifecycle, was 56.25% and 57.14%, respectively. The former project was better because it decreased the false positive rate at a lower running cost. CONCLUSION: The DT-LC is an effective screening strategy for second trimester detection of fetal Down's syndrome in mainland China. PMID- 21644236 TI - [Genetic polymorphism of the killer cell immunoglobulin-like receptor genes in the Inner Mongolian population]. AB - OBJECTIVE: To investigate the killer cell immunoglobulin-like receptor (KIR) gene frequencies and genotypes distributions in the Inner Mongolian population. METHODS: Ninety genomic DNA samples were extracted from blood samples of randomly chosen Mongolian individuals. Gene-specific PCR amplification was used to identify genes present or absent for 16 KIR loci. KIR genotype distributions were obtained and compared to that of 24 populations published in literatures using principal component analysis by SAS8.0 software. Genetic tree was constructed by the calculate Nei's genetic distance. RESULTS: (1) The frequency of KIR 2DL2, 2DS2 in Mongolian individual is higher than that in north Mongoloid and less than that in Caucasian. (2) Haplotype AA was identified in 37.78% of individuals, which is higher than that in north Mongoloid and lower than that in Caucasian. (3) Mongolian was considered between north Mongoloid and Caucasian by principal component and genetic tree analysis. CONCLUSION: Mongolian might be affected by the north Mongoloid and Caucasian, and showed intermediate between the two populations. PMID- 21644237 TI - [Distribution of HLA-C genes and HLA C-B, A-C-B haplotypes in Jinuo, Maonan and Wa ethnic populations in southwest China]. AB - OBJECTIVE: To investigate the distribution of human leukocyte antigen(HLA) class I genes and haplotypes in Jinuo, Maonan and Wa ethnic populations in southwest China. METHODS: Polymerase chain reaction-sequence specific oligonucleotide (PCR SSO) typing by Luminex was performed to genotype the HLA-C alleles in unrelated healthy individuals in the three populations. HLA C-B, A-C-B haplotypes were computed by combining the previous HLA-A and -B genotyping data using Pypop7.0 software. RESULTS: Eighteen HLA-C genes were identified in the three populations, with 17, 13 and 15 HLA-C genes in Jinuo, Maonan and Wa populations respectively. The alleles with frequency of more than 10% from high to low were C*08:01, C*01:02, C*03:04 and C*07:02 in the Jinuo, C*03:04, C*01:02, C*07:02 and C*08:01 in the Maonan, and C*12:03, C*08:01, C*07:02 and C*04:01 in the Wa. The predominant HLA A-C-B haplotypes were A*02:07-C*01:02-B*46:01, A*11:01-C*08:01 B*15:02 and A*11:01-C*03:04-B*13:01 in the Jinuo, A*11:01-C*03:04-B*13:01, A*02:07-C*01:02-B*46:01, A*11:01-C*08:01-B*15:02 and A*02:03-C*07:02-B*38:02 in the Maonan, and A*11:01-C*08:01-B*15:02, A*11:01-C*12:03-B*15:32 and A*11:01 C*04:01-B*35:01 in the Wa, respectively. CONCLUSION: There were different characteristics in the distributions of HLA-C genes and HLA C-B, A-C-B haplotypes in the Jinuo, Maonan and Wa populations. However, haplotypes C*08:01-B*15:02 and A*11:01-C*08:01-B*15:02 with high frequencies were common in the three populations, which might be the common ancient haplotypes of southern Chinese population. The study of HLA genes and haplotypes in these populations may be of significance in the study of population genetics, transplantation and disease association. PMID- 21644238 TI - Systematic review of immunohistochemical biomarkers to identify prognostic subgroups of patients with pancreatic cancer. AB - BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) carries a dismal prognosis. There is a need to identify prognostic subtypes of PDAC to predict clinical and therapeutic outcomes accurately, and define novel therapeutic targets. The purpose of this review was to provide a systematic summary and review of available data on immunohistochemical (IHC) prognostic and predictive markers in patients with PDAC. METHODS: Relevant articles in English published between January 1990 and June 2010 were obtained from PubMed searches. Other articles identified from cross-checking references and additional sources were reviewed. The inclusion was limited to studies evaluating IHC markers in a multivariable setting. RESULTS: Database searches identified 76 independent prognostic and predictive molecular markers implicated in pancreatic tumour growth, apoptosis, angiogenesis, invasion and resistance to chemotherapy. Of these, 11 markers (Ki 67, p27, p53, transforming growth factor beta1, Bcl-2, survivin, vascular endothelial growth factor, cyclo-oxygenase 2, CD34, S100A4 and human equilibrative nucleoside transporter 1) provided independent prognostic or predictive information in two or more separate studies. CONCLUSION: None of the molecular markers described can be recommended for routine clinical use as they were identified in small cohorts and there were inconsistencies between studies. Their prognostic and predictive values need to be validated further in prospective multicentre studies in larger patient populations. A panel of molecular markers may become useful in predicting individual patient outcome and directing novel types of intervention. PMID- 21644239 TI - Systematic review of intraperitoneal chemotherapy for gastric cancer. AB - BACKGROUND: Gastric cancer with peritoneal carcinomatosis has a poor prognosis. Intraperitoneal chemotherapy has been proposed as a treatment option. This systematic review examined recent literature to determine the role of intraperitoneal chemotherapy in gastric cancer. METHODS: Four electronic databases were searched between January 2004 and January 2010 for relevant studies. Defined outcomes of interest were treatment-related morbidity and mortality, long-term survival and sites of recurrence. RESULTS: Fourteen studies were identified involving 914 patients with gastric cancer, of whom 819 (89.6 per cent) received intraperitoneal chemotherapy. There were two randomized controlled trials, two case-control studies and ten observational studies. Methodological quality was rated as poor in 12 studies, with selection and observer bias apparent in most non-randomized cohorts. Studies were often small and varied in terms of intraperitoneal timing of chemotherapy, chemotherapeutic agents, treatment temperature, and the use of adjuvant therapies. In the better conducted studies, survival was longer in patients receiving intraperitoneal chemotherapy and surgery than in those having surgery alone. CONCLUSION: There is limited good quality evidence to determine the role of intraperitoneal chemotherapy in gastric cancer. Intraperitoneal chemotherapy in gastric cancer is worthy of further appraisal. However, the quality of trials must be improved, and studies must be conducted more uniformly to minimize bias and aid comparison between centres. PMID- 21644240 TI - Benzylidene acetal type bridged nucleic acids: changes in properties upon cleavage of the bridge triggered by external stimuli. AB - Four classes of benzylidene acetal type bridged nucleic acids (BA-BNAs) were designed with 2',4'-bridged structures that cleaved upon exposure to appropriate external stimuli. Cleavage of 6-nitroveratrylidene and 2-nitrobenzylidene acetal type BNA bridges occurred upon photoirradiation and subsequent treatment with thiol caused changes in secondary structure to afford 4'-C-hydroxymethyl RNA. Benzylidene and 4-nitrobenzylidene acetal type BNA responded to acids and reducing agents, respectively, resulting in hydrolysis of the acetal-bridged structure. Cleavage of the bridge removed sugar conformational restrictions and changed the duplex- and triplex-forming properties of the BNA-modified oligonucleotides. Moreover, oligonucleotides incorporating a single BA-BNA modification had considerably improved stability toward 3'-exonuclease, which was lost upon cleavage of the bridge. Thus, these new BNAs may be useful as therapeutic and detection tools by sensing various environments. PMID- 21644241 TI - Polymer brushes showing non-fouling in blood plasma challenge the currently accepted design of protein resistant surfaces. AB - Ultra-low-fouling poly[N-(2-hydroxypropyl) methacrylamide] (poly(HPMA)) brushes have been synthesized for the first time. Similar to the so far only ultra-low fouling surface, poly(carboxybetaine acrylamide), the level of blood plasma fouling was below the detection limit of surface plasmon resonance (SPR, 0.03 ng.cm(-2)) despite being a hydrogen bond donor and displaying a moderate wettability, thus challenging the currently accepted views for the design of antifouling properties. The antifouling properties were preserved even after two years of storage. To demonstrate the potential of poly(HPMA) brushes for the preparation of bioactive ultra-low fouling surfaces a label-free SPR immunosensor for detection of G Streptococcus was prepared. PMID- 21644242 TI - Controllable optical, electrical, and morphologic properties of 3,4 ethylenedioxythiophene based electrocopolymerization films. AB - This contribution presents a kind of novel and neutral network films based on EDOT formed by in situ electrocopolymerization (ECP). The ECP films which are neutral and colorless exhibit the conductivity of 0.2-0.5 S.cm(-1), W(F) of 4.79 5.20 eV, and RMS roughness of 3.51-5.26 nm. The electroluminescent devices where ECP films acted as hole-transport layer (HTL) exhibit higher brightness, current density, efficiency (20-30% improvement), and stability than that of PEDOT:PSS HTL device. The ECP films also significantly benefit the stability of neighboring organic layer compared to PEDOT:PSS. This kind of new ECP films affords more opportunities to develop organic light-emitting diodes (OLEDs) with high performances and stability. PMID- 21644243 TI - Discrete regulatory regions control early and late expression of D-Pax2 during external sensory organ development. AB - The transcription factor D-Pax2 is required for the correct differentiation of several cell types in Drosophila sensory systems. While the regulation of its expression in the developing eye has been well studied, little is known about the mechanisms by which the dynamic pattern of D-Pax2 expression in the external sensory organs is achieved. Here we demonstrate that early activation of D-Pax2 in the sensory organ lineage and its maintenance in the trichogen and thecogen cells are governed by separate enhancers. Furthermore, the initial activation is controlled in part by proneural proteins whereas the later maintenance expression is regulated by a positive feedback loop. PMID- 21644244 TI - An improved synthesis of (2S, 4S)- and (2S, 4R)-2-amino-4-methyldecanoic acids: assignment of the stereochemistry of culicinins. AB - An improved synthesis of (2S, 4S)- and (2S, 4R)-2-amino-4-methyldecanoic acids was accomplished using a glutamate derivative as starting material and Evans' asymmetric alkylation as the decisive step. The NMR data of the two diastereomers were measured and compared with those of the natural product. As a result, the stereochemistry of this novel amino acid unit in culicinins was assigned as (2S, 4R). PMID- 21644245 TI - Cooperativity network of Trp-cage miniproteins: probing salt-bridges. AB - Trp-cage miniprotein was used to investigate the role of a salt-bridge (Asp(9) Arg(16) ) in protein formation, by mutating residues at both sides, we mapped its contribution to overall stability and its role in folding mechanism. We found that both of the above side-chains are also part of a dense interaction network composed of electrostatic, H-bonding, hydrophobic, etc. components. To elucidate the fold stabilizing effects, we compared and contrasted electronic circular dichroism and NMR data of miniproteins equipped with a salt-bridge with those of the salt-bridge deleted mutants. Data were acquired both in neutral and in acidic aqueous solutions to decipher the pH dependency of both fully and partially charged partners. Our results indicate that the folding of Trp-cage miniproteins is more complex than a simple two-state process as we detected an intermediate state that differs significantly from the native fold. The intermediate formation is related to the salt-bridge stabilization; in the miniprotein variants equipped with salt-bridge the population of the intermediate state at acidic pH is significantly higher than it is for the salt-bridge deleted mutants. In this molecular framework Arg(16) stabilizes more than Asp(9) does, because of its higher degree of 3D-fold cooperation. In conclusion, the Xxx(9) leftright arrow Xxx(16) salt-bridge is not an isolated entity of this fold; rather it is an integrated part of a complex interaction network. PMID- 21644246 TI - Synthetic studies toward labionin, a new alpha,alpha-disubstituted amino acid from type III lantibiotic labyrinthopeptin A2. AB - The labyrinthopeptins are a new class of lantibiotics containing two identical quaternary alpha,alpha-disubstituted amino acids, named labionin (Lab). The synthetic formation of this unique structural feature represents the key step in the total synthesis of these polycyclic peptides. In this report we describe the synthesis of an orthogonally protected alpha,alpha-disubstituted amino acid building block serving as labionin precursor for the future assembly of labyrinthopeptin A2 and of other labyrinthopeptin derivatives. PMID- 21644247 TI - Creatinyl amino acids: new hybrid compounds with neuroprotective activity. AB - Prolonged oral creatine administration resulted in remarkable neuroprotection in experimental models of brain stroke. However, because of its polar nature creatine has poor ability to penetrate the blood-brain barrier (BBB) without specific creatine transporter (CRT). Thus, synthesis of hydrophobic derivatives capable of crossing the BBB by alternative pathway is of great importance for the treatment of acute and chronic neurological diseases including stroke, traumatic brain injury and hereditary CRT deficiency. Here we describe synthesis of new hybrid compounds-creatinyl amino acids, their neuroprotective activity in vivo and stability to degradation in different media. The title compounds were synthesized by guanidinylation of corresponding sarcosyl peptides or direct creatine attachment using isobutyl chloroformate method. Addition of lipophilic counterion (p-toluenesulfonate) ensures efficient creatine dissolution in DMF with simultaneous protection of guanidino group towards intramolecular cyclization. It excludes the application of expensive guanidinylating reagents, permits to simplify synthetic procedure and adapt it to large-scale production. The biological activity of creatinyl amino acids was tested in vivo on ischemic stroke and NaNO(2) -induced hypoxia models. One of the most effective compounds creatinyl-glycine ethyl ester increases life span of experimental animals more than two times in hypoxia model and has neuroprotective action in brain stroke model when applied both before and after ischemia. These data evidenced that creatinyl amino acids can represent promising candidates for the development of new drugs useful in stroke treatment. PMID- 21644248 TI - Two antimicrobial and nematicidal peptides derived from sequences encoded Picea sitchensis. AB - Two antimicrobial peptides (piceain 1 and 2) derived from sequences encoded Picea sitchensis are identified. Their amino acid sequences are KSLRPRCWIKIKFRCKSLKF and RPRCWIKIKFRCKSLKF, respectively. One intra-molecular disulfide bridge is formed by these two half-cysteines in both piceain 1 and 2. Antimicrobial activities of synthesized piceains against several kinds of microorganisms were tested. They showed antimicrobial activities against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and fungus Candida albicans but little antimicrobial activity against Bacillus subtilis. The results of nematicidal test showed they exerted strong nematicidal activities against Caenorhabditis elegans, following exposure for 5 h at concentrations as low as 10 ug/ml. They had weak hemolytic abilities against human and rabbit red cells. At the concentration of 250 ug/ml, they induced red cell hemolysis of less than 5%. Circular dichroism spectra of the two antimicrobial peptides were investigated in several solutions. Their main secondary structure components are beta-sheet and random. The current work provides a novel family of antimicrobial and nematicidal peptides with unique disulfided loop containing nine amino acid residues. PMID- 21644249 TI - Increasing the hydrolysis constant of the reactive site upon introduction of an engineered Cys14-Cys39 bond into the ovomucoid third domain from silver pheasant. AB - P14C/N39C is the disulfide variant of the ovomucoid third domain from silver pheasant (OMSVP3) introducing an engineered Cys14-Cys39 bond near the reactive site on the basis of the sequence homology between OMSVP3 and ascidian trypsin inhibitor. This variant exhibits a narrower inhibitory specificity. We have examined the effects of introducing a Cys14-Cys39 bond into the flexible N terminal loop of OMSVP3 on the thermodynamics of the reactive site peptide bond hydrolysis, as well as the thermal stability of reactive site intact inhibitors. P14C/N39C can be selectively cleaved by Streptomyces griseus protease B at the reactive site of OMSVP3 to form a reactive site modified inhibitor. The conversion rate of intact to modified P14C/N39C is much faster than that for wild type under any pH condition. The pH-independent hydrolysis constant (K(hyd) degrees ) is estimated to be approximately 5.5 for P14C/N39C, which is higher than the value of 1.6 for natural OMSVP3. The reactive site modified form of P14C/N39C is thermodynamically more stable than the intact one. Thermal denaturation experiments using intact inhibitors show that the temperature at the midpoint of unfolding at pH 2.0 is 59 degrees C for P14C/N39C and 58 degrees C for wild type. There have been no examples, except P14C/N39C, where introducing an engineered disulfide causes a significant increase in K(hyd) degrees , but has no effect on the thermal stability. The site-specific disulfide introduction into the flexible N-terminal loop of natural Kazal-type inhibitors would be useful to further characterize the thermodynamics of the reactive site peptide bond hydrolysis. PMID- 21644250 TI - beta-Amino acids containing peptides and click-cyclized peptide as beta-turn mimics: a comparative study with 'conventional' lactam- and disulfide-bridged hexapeptides. AB - The increasing interest in click chemistry and its use to stabilize turn structures led us to compare the propensity for beta-turn stabilization of different analogs designed as mimics of the beta-turn structure found in tendamistat. The beta-turn conformation of linear beta-amino acid-containing peptides and triazole-cyclized analogs were compared to 'conventional' lactam- and disulfide-bridged hexapeptide analogs. Their 3D structures and their propensity to fold in beta-turns in solution, and for those not structured in solution in the presence of alpha-amylase, were analyzed by NMR spectroscopy and by restrained molecular dynamics with energy minimization. The linear tetrapeptide Ac-Ser-Trp-Arg-Tyr-NH(2) and both the amide bond-cyclized, c[Pro-Ser Trp-Arg-Tyr-D-Ala] and the disulfide-bridged, Ac-c[Cys-Ser-Trp-Arg-Tyr-Cys]-NH(2) hexapeptides adopt dominantly in solution a beta-turn conformation closely related to the one observed in tendamistat. On the contrary, the beta-amino acid containing peptides such as Ac-(R)-beta(3) -hSer-(S)-Trp-(S)-beta(3) -hArg-(S) beta(3) -hTyr-NH(2) , and the triazole cyclic peptide, c[Lys-Ser-Trp-Arg-Tyr betatA]-NH(2) , both specifically designed to mimic this beta-turn, do not adopt stable structures in solution and do not show any characteristics of beta-turn conformation. However, these unstructured peptides specifically interact in the active site of alpha-amylase, as shown by TrNOESY and saturation transfer difference NMR experiments performed in the presence of the enzyme, and are displaced by acarbose, a specific alpha-amylase inhibitor. Thus, in contrast to amide-cyclized or disulfide-bridged hexapeptides, beta-amino acid-containing peptides and click-cyclized peptides may not be regarded as beta-turn stabilizers, but can be considered as potential beta-turn inducers. PMID- 21644251 TI - Identification of volatiles from pineapple (Ananas comosus L.) pulp by comprehensive two-dimensional gas chromatography and gas chromatography/mass spectrometry. AB - Combining qualitative data from the chromatographic structure of 2-D gas chromatography with flame ionization detection (GC*GC-FID) and that from gas chromatography-mass spectrometry (GC/MS) should result in a more accurate assignment of the peak identities than the simple analysis by GC/MS, where coelution of analytes is unavoidable in highly complex samples (rendering spectra unsuitable for qualitative purposes) or for compounds in very low concentrations. Using data from GC*GC-FID combined with GC/MS can reveal coelutions that were not detected by mass spectra deconvolution software. In addition, some compounds can be identified according to the structure of the GC*GC-FID chromatogram. In this article, the volatile fractions of fresh and dehydrated pineapple pulp were evaluated. The extraction of the volatiles was performed by dynamic headspace extraction coupled to solid-phase microextraction (DHS-SPME), a technique appropriate for slurries or solid matrices. Extracted analytes were then analyzed by GC*GC-FID and GC/MS. The results obtained using both techniques were combined to improve compound identifications. PMID- 21644252 TI - Characterization of the human serum depletome by label-free shotgun proteomics. AB - While it is known that immunoaffinity depletion of abundant proteins in serum removes additional proteins beyond those targeted, there has been little characterization of the co-depleted proteins in the high abundant fraction, which we refer to here as the "depletome". We present evidence of co-depletion of non targeted proteins in human serum using a top-20 immunodepletion column, as shown by label-free liquid chromatography mass spectrometry (LC-MS(E)) profiling. This led to identification of 147 proteins which were specific for this fraction and comprised proteins with functions predominantly in binding and transport of nucleotides, metal ions, carbohydrates and lipids. These results suggest that further studies on this commonly ignored serum fraction may provide new insights into clinical proteomics. PMID- 21644253 TI - Simultaneous analysis of acetaminophen, p-aminophenol and aspirin metabolites by hydrophilic interaction and strong anion exchange capillary liquid chromatography coupled to amperometric detection. AB - A simple and sensitive method has been developed for the simultaneous determination of polar nonsteroidal pharmaceuticals and metabolites, including acetaminophen, p-aminophenol and several aspirin metabolites (salicylic acid, gentisic acid, salicyluric acid and 2,3-dihydroxybenzoic acid), by capillary liquid chromatography with amperometric detection. Using a capillary monolithic column with mixed mode stationary phases and a mobile phase composed of acetonitrile and Tris buffer, rapid separation of six polar analytes was achieved within 8 min, and a hydrophilic interaction and strong anion exchange separation mechanism were exhibited. Method detection limits of six analytes ranged from 10 to 50 ng/mL. In terms of precision, the intra- and interday relative standard deviation values in all analytes never exceeded 3.1% for migration time and 8.9% for peak areas, respectively. This method provided a simple, rapid and cost effective approach for the analysis of polar pharmaceuticals. The applicability of the method in pharmacokinetics was verified by spiking human serum samples with the compounds and analyzing the recoveries. PMID- 21644254 TI - Development and validation of an HPLC method for the determination of ten sulfonamide residues in milk according to 2002/657/EC. AB - A HPLC method with diode-array detection, at 265 nm, was developed and validated for the determination of ten sulfonamides (SAs): sulfadiazine (SDZ), sulfathiazine (STZ), sulfamethoxine (SMTH), sulfamethizole (SMZ), sulfamethoxypyridazine (SMPZ), sulfamonomethoxine (SMMX), sulfamethoxazole (SMXZ), sulfisoxazole (SIX), sulfadimethoxine (SDMX), and sulfaquinoxaline (SQX) in milk. A mixture of ethyl acetate, n-hexane, and isopropanol was used for the extraction of target analytes from milk. The mobile phase, a mixture of 0.1% v/v formic acid, CH(3) CN, and CH(3) OH was delivered to the analytical column under a gradient program. The procedure was validated according to the European Union regulation 2002/657/EC in terms of selectivity, stability, decision limit, detection capability, accuracy, and precision. Mean recoveries of sulfonamides from milk samples spiked at three concentration levels (0.5*MRL, 1*MRL, and 1.5*MRL) (MRL, maximum residue level) were 93.9-115.9% for SDZ, 97.8-102.9% for STZ, 94.6-107.0% for SMTH, 98.3-111.5% for SMZ, 95.3-108.4% for SMPZ, 97.9-106.0% for SMMX, 97.6-111.3% for SMXZ, 94.3-104.6% for SIX, 96.4-109.1% for SDMX, and 98.2-111.2% for SQX. All RSD values were lower than 8.8%. The decision limits CCa calculated by spiking 20 blank milk samples at MRL (100 MUg/kg) ranged from 101.61 to 106.84 MUg/kg, whereas the detection capability CCb ranged from 105.64 to 119.01 MUg/kg. PMID- 21644255 TI - Kinetic characterization of the group II Helicoverpa armigera nucleopolyhedrovirus propagated in suspension cell cultures: implications for development of a biopesticides production process. AB - Large-scale commercialization of baculovirus biopesticides for the control of insect pests requires a cell culture production process, and knowledge of the infection kinetics is a vital prerequisite for process optimization. Well characterized kinetic parameters have so far only been reported for the commercially established recombinant Autographa californica nucleopolyhedrovirus (AcMNPV), a Group I NPV. In this work, key infection kinetic parameters of the Group II NPV Helicoverpa armigera nucleopolyhedrovirus (HaSNPV), and its Few Polyhedra (FP) mutant, were well characterized for the first time, in suspension HzAM1 insect cell cultures, to facilitate the scale-up of an HaSNPV-based biopesticide. The FP mutant had a selective advantage over wild-type HaSNPV in cell cultures, and the kinetic analysis showed that this was due to a superior budding rate, rather than a faster binding rate (BR) or longer budding duration. Another finding was that wild-type HaSNPV had very poor infection kinetics when compared with AcMNPV, exhibiting an 18-fold lower BR, a more than 50-fold lower budding rate, and a 60-fold lower extracellular/total progeny virus ratio. Such poor infection kinetics have serious implications during scale-up of an HaSNPV biopesticide production process, including the requirement for large volumes of virus inocula and the difficulty of achieving synchronous infections. Groups I and II NPVs may have very different infection kinetics because of their different envelope fusion proteins. This study is the first to compare the two groups of NPVs in terms of well-characterized cell-specific infection kinetics, and the findings may indicate a phylogenetic basis for kinetic differences. PMID- 21644256 TI - [Hospices - a place to die with dignity]. PMID- 21644257 TI - [At least three good reasons to speak]. PMID- 21644258 TI - [Thanks to education, patients may refuse programming of their social destiny and improve their health]. PMID- 21644259 TI - [Neurology for the general practitioners. Assessing and stopping vertigo]. PMID- 21644260 TI - [Self-help-friendly doctors' practices--recent approaches to promote collaboration between physicians and self-help groups]. PMID- 21644261 TI - Unseasonal outbreaks of husk in dairy cattle. PMID- 21644262 TI - Equine disease surveillance: quarterly summary. PMID- 21644263 TI - Melanoma cell-secreted soluble factor that stimulates ubiquitination and degradation of the interferon alpha receptor and attenuates its signaling. PMID- 21644264 TI - [Cell repair: break the pathway]. PMID- 21644265 TI - [Chromotripsy]. PMID- 21644266 TI - Numerous reports of metabolic disease in dairy cows. PMID- 21644267 TI - Back to the bench. PMID- 21644268 TI - Conflicted!Part I: by Caveman. PMID- 21644269 TI - Well-known voice from MLO's past. PMID- 21644271 TI - Get out that crystal ball. PMID- 21644270 TI - It may not be "a little dementia". PMID- 21644272 TI - [Cancer and obesity: effect of bariatric surgery]. PMID- 21644273 TI - [European Resuscitation Council guidelines for resuscitation 2010]. AB - BASIC LIFE SUPPORT: All rescuers trained or not, should provide chest compressions to victims of cardiac arrest. The aim should be to push to a depth of at least 5 cm at a rate of at least 100 compressions per minute, to allow full chest recoil, and to minimise interruptions in chest compressions. Trained rescuers should also provide ventilations with a compression-ventilation ratio of 30:2. ELECTRICAL THERAPIES: Much greater emphasis on minimising the duration of the pre-shock and post-shock pauses; the continuation of compressions during charging of the defibrillator is recommended. Further development of AED programmes is encouraged. ADULT ADVANCED LIFE SUPPORT: Increased emphasis on high quality chest compressions throughout any ALS intervention paused briefly only to enable specific interventions. Removal of the recommendation for a pre-specified period of cardiopulmonary resuscitation before out-of-hospital defibrillation following cardiac arrest unwitnessed by the EMS. The role of precordial thump is de-emphasized. Delivery of drugs via a tracheal tube is no longer recommended, drugs should be given by the intraosseous (IO) route. Atropine is no longer recommended for routine use in asystole or pulseless electrical activity. Reduced emphasis on early tracheal intubation unless achieved by highly skilled individuals with minimal interruptions in chest compressions. Increased emphasis on the use of capnography. Recognition of potential harm caused by hyperoxaemia. Revision of the recommendation of glucose control. Use of therapeutic hypothermia to include comatose survivors of cardiac arrest associated initially with shockable rhythms, as well as non-shockable rhythms, with a lower level of evidence acknowledged for the latter. INITIAL MANAGEMENT OF ACUTE CORONARY SYNDROMES: The term non-ST-elevation myocardial infarction-acute coronary syndrome (non-STEMI-ACS) has been introduced for both NSTEMI and unstable angina pectoris. Primary PCI (PPCI) is the preferred reperfusion strategy provided it is performed in a timely manner by an experienced team. Non-steroidal anti inflammatory drugs should be avoided, as well as routine use of intravenous beta blockers; oxygen is to be given only to those patients with hypoxaemia, breathlessness or pulmonary congestion. PAEDIATRIC LIFE SUPPORT: The decision to begin resuscitation must be taken in less than 10 seconds. Lay rescuers should be taught to use a ratio of 30 compressions to 2 ventilations, rescuers with a duty to respond should learn and use a 15:2 ratio; however, they can use the 30:2 compression-ventilation ratio if they are alone. Ventilation remains a very important component of resuscitation in asphyxial arrest. The emphasis is on achieving quality compressions with the rate of at least 100 but not greater than 120 per minute, with minimal interruptions. AEDs are safe and successful when used in children older than 1 year. A single shock strategy using a non escalating dose of 4 J/kg is recommended for defibrillation in children. Cuffed tubes can be used safely in infants and young children. Monitoring exhaled carbon dioxide (CO2), ideally by capnography, is recommended during resuscitation. RESUSCITATION OF BABIES AT BIRTH: For uncompromised babies, a delay in cord clamping of at least one minute from the complete delivery is now recommended. For term infants, air should be used fro resuscitation at birth. For preterm babies less than 32 weeks gestation blended oxygen and air should be given judiciously and its use guided by pulse oximetry. Preterm babies of less than 28 weeks gestation should be completely covered in a plastic wrap up to their necks, without drying, immediately after birth. The recommended compression: ventilation ratio remains at 3:1 for newborn resuscitation. Attempts to aspirate meconium from the nose and mouth of the unborn baby, while the head is still on the perineum, are not recommended. If adrenaline is given the n the intravenous route is recommended using a dose of 10-30 microg/kg. Newly born infants born at term or near-term with moderate to severe hypoxic-ischaemic encephalopathy should be treated with therapeutic hypothermia. PRINCIPLES OF EDUCATION IN RESUSCITATION: The aim is to ensure that learners acquire and retain skill and knowledge that will enable them to act correctly in actual cardiac arrest and improve patient outcome. Short video/computer self-instruction courses, with minimal or no instructor coaching, combined with hands-on practice can be considered as an effective alternative to instructor-led basic life support (BLS and AED) courses. Ideally all citizens should be trained in standard CPR that includes compressions and ventilations. Basic and advanced life support knowledge and skills deteriorate in as little as three to six months. CPR prompt or feedback devices improve CPR skill acquisition and retention. PMID- 21644274 TI - [Decrease of urinary tract infections following catheterization after the education of health care workers, introduction of protocols and surveillance lists]. AB - Urinary tract infections (UTI) following catheterization are the most common hospital-acquired infections, with their frequency amounting to 30-40% of all hospital infections. Major percentage of this kind of infectious episodes can be prevented via active and continual education of health care workers (HCW), implementation of transparent protocols concerning installation and attendance of urinary catheters and regular control of catheterized patients through surveillance lists. This research shows the importance of a fore-mentioned activities, demonstrating a significant decrease of UTI following catheterization at the Department of Urology in Slavonski Brod General Hospital during two periods. After the aforementioned procedures were conducted, a statistically significant discrepancy in the decrease of the UTI incidence following catheterization was identified, from 20.4% to 11.7%, i.e. chi2 = 17.5; p < 0.01, and accordingly, significant decrease of the number of hospital bed-days, i.e. chi2 = 16.62; p < 0.0, while total consumption of antibiotics at the Department was not reduced, despite the decrease in the number of UTI. The most common uropathogens, with no significant difference in both periods, were E. coli (29.7%), Enterococcus spp (20%), and Pseudomonas aeruginosa (15.8%). Strict implementation and close surveillance of the recommended preventive measures are an important factor in reducing the number of hospital infections. PMID- 21644275 TI - [Surgical treatment of hallux valgus with new method: operative technique and review of 100 cases]. AB - We present the operative technique and treatment results for a new three dimensional method for hallux valgus correction. Lucijanic procedure was developed at the Department of Orthopaedic Surgery, General Hospital Karlovac, where patients enrolled in this study were treated from 2001 to 2008. Clinical and radiological evaluation was performed in 100 cases with a mean follow-up of 4.5 years. Mean American Orthopaedic Foot and Ankle Society score improved from 48.56 preoperatively to 92.34 points postoperatively. Average recovery time and return to work was 7.7 weeks. Result was excellent or good in 93%, fair in 5% and poor in 2% of feet. On the average hallux valgus angle decreased from 29.7 degrees to 9.3 degrees and on the average intermetatarsal angle decreased from 12.5 degrees to 5.2 degrees. First metatarsal inclination angle on average increased 5.3 degrees. First metatarsophalangeal joint congruence and tibial sesamoid position were corrected. The new method allows for correction of hallux valgus deformity in all three planes and for metatarsalgia attenuation. PMID- 21644276 TI - [Relapsing polychondritis--case report]. AB - Relapsing polychondritis (RP) is a rare systemic inflammatory disease in which recurrent episodes of cartilage inflammation result in destruction of ears, nose and tracheobronchal tract. The joints, eyes, audiovestibular system and cardiovascular system can also be involved. About 30% of patients with RP have coexisting autoimmune disease, or malignant disease like colon, breast, and lung carcinoma, or malignant lymphoma. Pathogenesis is still unknown, and there is no consistent laboratory parameter specific for RP, which makes the diagnosis mainly clinical. Glucocorticoids are a mainstay of medical treatment of RP, whereas newer studies show positive effects of biological therapy. The course of RP is characterized by recurrent episodes of cartilage inflammation, and the prognosis has been recently improved because of improved medical and surgical treatment. We present a case of a patient with RP who was diagnosed 1 month after the development of first symptoms and responded well to glucocorticoid therapy. PMID- 21644277 TI - [Endoscopic transsphenoidal pituitary surgery]. AB - Endoscopic surgery has been performed in our Department since 1996 as assistance in micro-neurosurgical procedures. In the same way the endoscope was used in transsphenoidal pituitary surgery, but from the beginning of 2004 we started with "pure" endoscopic surgery. We present our experience with the first ninety patients to demonstrate our way of operating. Endoscopic pituitary surgery is performed through a natural nasal air pathway without any incisions. A 4-mm endoscope is placed in front of the tumor in the sphenoidal sinus and the tumor is removed with specially designed surgical tools. Postoperative nasal packing is not necessary and postoperative discomfort is minimal so the hospital stay lasts 3 days. The first control was after one month. There were eighteen recurrences of tumors. PMID- 21644278 TI - [Vitamin B12 deficiency in children--underestimated danger in the light of new knowledge]. AB - Vitamin B12 (cobalamin) has two active forms, adenosylcobalamin and methylcobalamin which have a key role in two important metabolic pathways in humans and their deficiency is responsible for clinical problems. Cobalamin is essential during whole life, but its sufficient amount is extra important in fetal and neonatal period, when it is essential for normal child growth and development as well as for normal development of the central nervous system. Because of very complex transport and metabolism, its deficiency can be manifested in numerous congenital and acquired disorders. Vitamin B12 deficiency mostly has non-specific clinical features, it carries a great risk of permanent consequences, but most frequently it is easily curable if diagnosed on time. In Croatia cobalamin deficiency in children has been diagnosed too rarely. Accordingly, the aim of this paper is to point to the recently gained knowledge on cobalamin metabolism, present typical case reports and to provide guidelines for rapid and proper diagnostic and therapeutic approach. PMID- 21644279 TI - [Vertebrobasilar insufficiency and obstructive sleep apnea]. AB - The aim of this article is to point to correlation between vertebrobasilar insufficiency (VBI) and obstructive sleep apnea (OSA) that has not been perceived sufficiently till now. Namely, in the voluminous literature about sleep disordered breathing, VBI has been cited only as one of the possible causes of central and mixed sleep apnea. However, according to our clinical observation, VBI could be one of the most important factors in etiopathogenesis of OSA. Results of recent research, which confirm the correlation between VBI and OSA, contribute to our hypothesis. High prevalence and pathogenesis of OSA in patients with VBI can be explained by hypoxia of the medulla oblongata during sleep (sleep+positional VBI --> hypoxia of the medulla oblongata --> hypoxia of the respiratory centre which regulates breathing function+hypoxia of the motor nuclei of the IX, X and XII cranial nerves whose neurons regulate the tone in upper airway muscles (IX, X) and tongue (XII) --> OSA). The development of OSA in certain patients with VBI is probably in close correlation with the degree of hypoxia of the medulla oblongata during sleep (moderate VBI --> OSA; severe VBI - > central or mixed sleep apnea). Considering the fact that VBI of vascular and/or compressive etiology can be the primary cause of OSA, the Doppler sonography examination ofVB system (VBS) with the positional functional tests should be included in the diagnostic algorithm for OSA. Since the functional and organic disorders of cervical spine can either cause circulation disturbances in VBS or aggravate insufficient circulation, especially during sleep in certain head and neck positions, detailed examination of cervical spine including physiatric examination, manual functional examination and radiological examination, should become a part of the routine examination in patients with either suspected or confirmed OSA. Circulation in VBS can be improved or normalized by an appropriate therapy for vascular disorders (for example, stenting of vertebral artery), and the compressive effect of cervical spine disorders on circulation in VBS can be reduced or eliminated by an appropriate therapy of cervical spine disorders (for example, manual therapy, physical therapy, kinesitherapy, surgery etc.). OSA symptoms can be significantly reduced or completely cease after an improvement or normalization of circulation in VBS. PMID- 21644280 TI - [Clinical and etiopathogenetic role of plasminogen and metaloproteinase systems in the tumor growth. Pericellular proteolysis of extracellular matrix and tumor growth]. AB - Pericellular proteolysis is a cascade process involved in degradation of extracellular matrix. This process is included in various physiological and pathological processes. Pericellullar proteolysis has major functions like degradation of tissue stroma and weakening of intercellular connections but it also has a function in the synthesis of bioactive molecules (cytokines, growth factors and inhibitory factors). Plasminogen system is involved in fibrinolysis and starts metalloproteinase activation. Activity of proteolytic molecules is controlled by the rate of zymogenic activation, half-life of molecules, and action of inhibitory molecules. Inhibition is achieved through direct binding of inhibitor and enzyme and takes a few steps. Pericellular proteolysis is involved in tumor invasion and metastasis, inflammatory reaction, degenerative diseases and other diseases. Pathophysiological regulation of pericellular proteolysis in mentioned diseases contributes to clinical properties of diseases and has diagnostic and therapeutic importance. PMID- 21644281 TI - [Five "W" in tuberculosis contact tracing: why, when, who, where, and what?]. AB - The most successful intervention in tuberculosis control is early detection and early and effective treatment of tuberculosis patient. Therefore, identification of secondary cases and their prompt treatment may interrupt further transmission. This is the goal in the national tuberculosis programmes in high-incidence countries since extensive contact investigation is resource intensive. As countries approach the elimination phase of tuberculosis, more aggressive strategies are needed. Contact tracing is regarded as an effective intervention to identify and to treat recently infected persons and has become an essential component of the tuberculosis control and elimination strategies in most low incidence countries. These activities can lead to the exhausting of the reservoir of future cases by preventing reactivation. Croatia is a middle--incidence country with decreasing incidence rates moving towards low-incidence situation. Contact investigation is an important part of tuberculosis control efforts. This study described current contact tracing principles and practice. PMID- 21644282 TI - [Ten years after publishing results from The Heart Outcomes Prevention Evaluation Study--HOPE: implications on treatment with angiotensin converting enzyme inhibitors in clinical practice]. AB - Ten years ago results from The Heart Outcomes Prevention Evaluation Study were published. Those results had a great impact on position of ACE inhibitors in clinical practice. In the study, comparison of ramipril in daily dosage up to 10 mg and placebo in 9541 patients with high cardiovascular risk during 4.5 years of follow-up resulted in relative risk reduction for all primary end-points: cardiovascular mortality, myocardial infarction and stroke for 22% in patients treated with ramipril. Systolic blood pressure was reduced minimally for 3 mmHg. The effect was independent of age, gender, concomitant diseases or therapy. Extended follow-up of the patients for further 2.6 years showed prolonged beneficial effect of ramipril. Ramipril reduced relative risk for newly developed diabetes for 34%. Results of the study greatly influenced the guidelines for treatment of patients with cardiovascular diseases. This study proved benefits of ACE inhibitors in primary prevention in patients with high cardiovascular risk. PMID- 21644283 TI - [New kind of prevention--quartery prevention]. PMID- 21644284 TI - [Joint pain in a young Muslim woman. Covered up too much]. PMID- 21644285 TI - [Emergency consultation for an unconscious patient. How to bill according to EBM, according to GOA?]. PMID- 21644286 TI - [Functional or mental problem. What constipation type is your patient?]. PMID- 21644287 TI - [Trends in sexually transmitted diseases. Chlamydia and gonococci are often searched for in the wrong place]. PMID- 21644288 TI - [Naturopathy consultation. Functional cardiovascular disorders]. PMID- 21644289 TI - [Female urinary incontinence affects all age groups. Well-tried and innovative treatment]. PMID- 21644290 TI - [Diagnostics for female incontinence: two-stage assessment]. PMID- 21644291 TI - [Non-surgical treatment of female incontinence]. PMID- 21644292 TI - [Surgical treatment for female stress urinary incontinence]. PMID- 21644293 TI - [New-onset type 1 diabetes: what to do, differential diagnosis and prevention]. PMID- 21644294 TI - [Emergency checklist: ketoacidotic coma]. PMID- 21644295 TI - [Opiates in the treatment of tumor related pain]. PMID- 21644296 TI - [Analysis of the nominations of acupuncture and moxibustion of traditional Chinese medicine for inscription on the Representative List of the Intangible Cultural Heritage of Humanity]. AB - The definition of intangible cultural heritage and the Representative List of the Intangible Cultural Heritage of Humanity in Convention for Safeguarding of the Intangible Cultural Heritage is discussed. Nominations of elements should be prepared in accordence with the Guidelines provided in each section. The explaination methods and the determining process of the Nominations for Acupuncture and Moxibustion on the Representative List are analyzed, such as the name of the element, characteristics, identification and definition, value and safeguarding measures, photos and video of the element. The Nominations should be prepared according to the Convention and Guidelines closedly and focus on discussing the cultural, the content, the communities and individuals, safeguarding measures of element. PMID- 21644297 TI - [Thirty-two cases of obstructive sleep apnea-hypopnea syndrome treated by acupuncture combined with acupoint injection]. PMID- 21644298 TI - [Observation on immediate analgesic effect of acupuncture at Shiqizhui (EX-B 8) only or multi-acupoints in patients with dysmenorrhea: a randomized controlled trial]. AB - OBJECTIVE: To compare the different therapeutic effect between acupuncture at Shiqizhui (EX-B 8) only and multi acupoints on dysmenorrhea. METHODS: Thirty eight cases were randomly divided into a single acupoint group and a multi acupoints group, 19 cases in each group. The single acupoint group was treated by acupuncture at Shiqizhui (EX-B 8) only, and the multi-acupoints group by acupuncture at Shiqizhui (EX-B 8), Sanyinjiao (SP 6), Diji (SP 8), Ciliao (BL 32). They were all treated from the first day when sudden intense pain occurs, one time each day, for 3 days in each menstrual cycle, the treatment of three menstrual cycles. The therapeutic effect and Visual Analogue Scale (VAS) were compared and the score of general frequency and severity of dysmenorrhea by using Cox Menstrual Symptom Scale (CMSS) were evaluated. RESULTS: The cured rate was 68.4% (13/19) and the effective rate was 31.6% (6/19) in the single acupoint group, being similar to 78.9% (15/19) and 21.1% (4/19) in the multi-acupoints group (P > 0.05). VAS and the scores of general frequency and severity of dysmenorrhea were all significantly decreased after treatment in both groups (all P < 0.001), with no significant difference between the two groups (all P > 0.05). CONCLUSION: Acupuncture at Shiqizhui (EX-B 8) only can be as effective as selecting multi-acupoints to cure essential dysmenorrhea. PMID- 21644299 TI - [Thirty-two cases of sequelae of brain concussion treated by bloodletting at Jinjin (EX-HN 12), Yuye (EX-HN 13) combined with acupuncture at Sishencong (EX-HN 1)]. PMID- 21644300 TI - [Observation on therapeutic effect of Tongluo Guben needling method for lumbar disc herniation]. AB - OBJECT: To explore the better treatment of lumbar intervertebral disc herniation. METHODS: One hundred and thirty-two cases of lumbar intervertebral disc herniation were randomly divided into an observation group and a control group, sixty-six cases in each group. In observation group, Baihui (GV 20) penetrating to Taiyang (EX-HN 5), Jiaji (EX-B 2), Huantiao (GB 30) and Chengfu (BL 36) etc. on affect side were applied; in control group, Shenshu (BL 23), Dachangshu (BL 25), Huantiao (GB 30) and Chengfu (BL 36) etc. on affect side were applied. Once a day, 6 treatments made one session and one day break between courses. The curative effects after 4 and 8 courses and the recurrence rate that 6 months after treatment were observed; the pain scale evaluated by Visual Analogue Scale (VAS) and the Oswestry Disability Index (ODI) were observed. RESULTS: The cured rate and total effective rate were 44.3% (27/61) and 100.0% (61/61) in observation group, 26.7% (16/60) and 93.3% (56/60) in control group after 4 courses; there were significant differences between two groups (all P < 0.05). After 8 courses, the cured rate and total effective rate were no significant differences between two groups (all P > 0.05). In observation group, the recurrence rate was 9.4% (5/53) 6 months after treatment, which was lower than that (24.1%,13/54) in control group (P < 0.05). The VAS and ODI scale were significantly reduced (all P < 0.05) after treatment, and which in observation group were inferior to those in control group (all P < 0.01). CONCLUSION: The lumbar intervertebral disc herniation treated with puncture by opening meridian and consolidating origin is rapidly effected with low recurrence rate. PMID- 21644301 TI - [Acupuncture combined with blood-letting and cupping for periarthritis of shoulder]. PMID- 21644302 TI - [Observation on relations between onset seasons of peripheral facial paralysis and therapeutic courses and effect of acupuncture]. AB - OBJECTIVE: To observe the influences of seasonal factors on peripheral facial paralysis by acupuncture. METHODS: Four hundred cases of facial paralysis were divided into spring, summer, autumn and winter groups, 100 cases in each group. All these cases were treated by routine puncture. Fengchi (GB 20), Yifeng (TE 17), Qianzheng (Extra), Jiache (ST 6), and Dicang (ST 4), etc. were applied at affect side, once a day. 2 months observation was carried on to compare the clinical therapeutic effects and average courses. The facial symptoms, physical sign and functional activities were taken as observation indexes of therapeutic effect. RESULTS: The effect rate was 78.0% (78/100) in spring group, 82.0% (82/100) in summer group, 89.0% (89/100) in autumn group and 92.0% (92/100) in winter group; the effect rate in summer or autumn group was superior to those in spring group and in summer group (all P < 0.05); the average course was (47.6 +/- 22.3) days in spring group, (43.7 +/- 18.4) days in summer group, (31.5 +/- 11.3) days in autumn group and (22.6 +/-9.2) days in winter group, indicating the significant differences between groups except that between spring and summer group (all P < 0.01). The cured and markedly effective rate was 80.1% (161/201) for wind cold type, 53.5% (61/114) for wind heat type, and 36.5% (31/85) for damp heat type, indicating that it of wind cold type was superior to that of wind heat type or damp heat type (P < 0.001, P < 0.05). CONCLUSION: The syndrome distribution and courses of peripheral facial paralysis are different in different seasons, hence, the diseases should be treated according to attack time and syndromes. PMID- 21644303 TI - [Thirty-two cases of intractable facial paralysis treated by wheat-grain size cone moxibustion]. PMID- 21644304 TI - [Effect of different needle retaining times of electroacupuncture on trigeminal neuralgia]. AB - OBJECTIVE: To observe the different effect of different needle retaining times of electroacupuncture on tri geminal neuralgia. METHODS: Fifty-two cases were randomly divided into an observation group (26 cases) and a control group (26 cases). Electroacupuncture at four pairs of acupoints including Xiaguan (ST 7) and Dicang (ST 4), Jiache (ST 6) and Chengjiang (CV 24), Sibai (ST 2) and Yingxiang (LI 20) and acupoints selected according to differentiation were used in both groups. The needle-retaining time of every pair of acupoints was 30 min (120 min together) in observation group, and that was 10 min (40 min together) in control group. The treatment frequency was once a day. The effect was evaluated after 4 weeks. RESULTS: The cured rate of observation group (84.6%, 22/26) was better than that of control group (34.6%, 9/26) (P < 0.01), and the total effective rate of the former (100.0%, 26/26) was also better than that of the latter (80.8%, 21/26) (P < 0.05). CONCLUSION: The effect of needle-retaining time for 30 min at every pair of acupoints is better than that for 10 min in electroacupuncture on trigeminal neuralgia. PMID- 21644305 TI - [Impacts of Taichong (LR 3) on blood flow velocity in patients with vertebrobasilar insufficiency]. AB - OBJECTIVE: To observe the intervention on the disturbance of blood flow velocity in vertebral artery in patients with vertebrobasilar insufficiency (VBI) treated with acupuncture at Taichong (LR 3) so that the clinical evidences could be provided for the research of acupoint specificity. METHODS: One hundred cases of VBI were tested with Transcranial Doppler (TCD) and 43 vessels of low velocity of blood flow and 79 vessels of high velocity of blood flow were discovered. Additionally, 50 cases of normal people were selected in control group, including totally 100 vertebral arteries. The velocity changes in systolic period (Vs) of vertebral artery were observed before and after acupuncture at Taichong (LR 3). RESULTS; After acupuncture at Taichong (LR 3), Vs of vertebral artery in low velocity cases was increased apparently. Vs in 5 to 10 min after acupuncture and half a hour after needle withdrawal was different significantly in statistics as compared with Vs before acupuncture (both P < 0.01). Vs of vertebral artery in high velocity cases was reduced apparently. Vs in 5 to 10 min after acupuncture and half a hour after needle withdrawal was different significantly in statistics as compared with Vs before acupuncture (both P < 0.01). There was no significant difference in statistics in comparison before and after acupuncture in control group (P > 0.05). CONCLUSION: Acupuncture at Taichong (LR 3) improves blood supply in vertebral artery in the mode of dual regulation and rectifies the disturbance of vertebral artery blood flow in dynamics. PMID- 21644306 TI - [Comparison on therapeutic effect between surround needling plus thin cotton moxibustion and Western medicine for herpes zoster]. AB - OBJECTIVE: To compare the therapeutic effect between surround needling plus thin cotton moxibustion and western medicine for herpes zoster. METHODS: Eighty cases were randomly divided into a surround needling plus thin cotton moxibustion group (observation group, n = 42) and a western medicine group (control group, n = 38) with a randomized controlled method. The observation group was treated with surround needling around the herpes and thin cotton moxibustion on the surface of the herpes, and the control group with oral administration of Acyclovir tablet plus topical Acyclovir cream. The scores of pain, quantity of herpes, herpes color, anabrosis and exudation changes of herpes and the markedly effective rate in the two groups were compared before and after treatment. RESULTS: The markedly effective rate was 78.6% (33/42) in the observation group, which was superior to 39.5% in the control group (P < 0.05). All the symptom scores in both the groups were improved obviously after treatment (all P < 0.05), and the observation group was better than the control group (all P < 0.05). CONCLUSION: The therapeutic effect of surround needling plus thin cotton moxibustion on herpes zoster is superior to that of routine western medicine. PMID- 21644307 TI - [Plum blossom needles combined with cupping for dermatitis due to mosquito bites]. PMID- 21644308 TI - [Effect of heat-sensitive point moxibustion on BMD, S-AKP, U-Ca/Cr in patients with primary osteoporosis]. AB - OBJECTIVE: To compare the curative effects of primary osteoporosis treated with heat-sensitive point moxibustion and Gaitianli (Oyster Shell and Calcium Carbonate Chewable) tablets for oral administration and explore the treatment mechanism. METHODS: Sixty cases of primary osteoporosis were randomly divided into a heat-sensitive point moxibustion group (moxibustion group) and a Gaitianli tablets group (medication group), 30 cases in each group. In the moxibustion group, the heat sensitized points were searched around Zusanli (ST 36), Pishu (BL 20), Shenshu (BL 23) and Mingmen (GV 4) and treated by heat-sensitive point moxibustion; in medication group, Gaitianli tablets were taken by oral administration, 3 pills for once and 3 times a day. The curative effects, bone mineral density (BMD), alkaline phosphatase (S-AKP) and urinary calcium to creatinine ratio (U-Ca/Cr) in both groups were observed before and after treatment. RESULTS: The total effective rate was 86.7% (26/30) in moxibustion group, superior to that of 76.7% (23/30) in medication group (P < 0.05). After treatment, the BMD of lumbar vertebrae (L2-L4) mean was improved (P < 0.05), and the S-AKP and U-Ca/Cr were reduced (all P < 0.05); in medi cation group, the indexes above were no obvious changes (all P > 0.05). CONCLUSION: The therapeutic effect of primary osteoporosis treated with heat-sensitive point moxibustion is superior to that with Gaitianli tablets for oral administration. The mechanism is restraining bone resorption, increasing bone strength, keeping balance of bone metabolism, in order to increase bone mineral density and improve the clinical symptoms. PMID- 21644309 TI - [Needle sticking method]. AB - Needle sticking method, which can be combined with multiple needling techniques, has been attached with great importance in recent years by doctors in clinic. Combining with the clinical experiences, the authors expounded the needle sticking method through its unified concept, differences between needle sticking method, which was an acupuncture technique, and stuck needle which was an accident during acupuncture, selection of needles, manipulations, mechanism of treatment, range of application, attentions and advantages of popularization. It is held that the technique can be widely applied for treatment of acute and chronic diseases of various departments with filiform needles. Easy to be manipulated, understood and mastered, the technique is without side effect and valuable to be popularized. PMID- 21644310 TI - [Comparison on heat sensation degree of ginger-partition moxibustion and suspended moxibustion at different acupoints for different time]. AB - OBJECTIVE: To measure the threshold of feeling temperature, comfortable temperature and tolerant temperature to ginger-partition moxibustion and suspended moxibustion at different acupoints of healthy persons, and to instruct the clinical practice of moxibustion better. METHODS: Forty-three healthy students were divided into a ginger-partition moxibustion group (22 cases) and a suspended moxibustion group (21 cases) randomly. Shenshu (BL 23), Zhongwan (CV 12), Zusanli (ST 36) and Waiguan (TE 5) were used in both groups. The feeling temperature, comfortable temperature and tolerant temperature in each group were measured by thermometer. SPSS 18.0 was used to analyze the data. RESULTS: The feeling temperature, comfortable temperature and tolerant temperature rose in proper order and the difference was significant (all P < 0.05). The tolerant temperature of suspended moxibustion group ((40.69 +/- 1.85) degrees C) was lower than that of ginger-partition moxibustion group ((42.44 +/- 3.90) degrees C) (P < 0.05). The feeling temperature of ginger-partition moxibustion at Shengshu (BL 23) was 4-5 degrees C lower than that at other acupoints. CONCLUSION: The heat sensations of different acupoints to ginger partition moxibustion and suspended moxibustion are different at different stages. The threshold of heat sensation of ginger-partition moxibustion is higher than that of suspended moxibustion. There is a best temperature zone for moxibustion which is a little higher than normal temperature between comfortable temperature and tolerant temperature. To improve effect, doctors should try to prolong the effective time of this zone in clinic. As the limiting temperature of patients, tolerant temperature can provide reference for the use of moxibustion safely in clinic. PMID- 21644311 TI - [One hundred and eighteen cases of simple epistaxis in plateau area treated by blood-letting puncture and cupping at Dazhui (GV 14)]. PMID- 21644312 TI - [Effect of different anesthesia methods on immune function in patients of laparoscopic cholecystectomy in peri-operational period]. AB - OBJECTIVE: To compare the influence of acupuncture combine with general anesthesia (ACGA) and general anesthesia (GA) on immune function in patients of laparoscopic cholecystectomy (LC) in peri-operational period. METHODS: Thirty nine cases undergoing LC were randomly divided into an ACGA group and a GA group. The ACGA group was treated with electroacupuncture at Hegu (LI 4), Zusanli (ST 36) and Yanglingquan (GB 34) for 15-30 minutes and subsequently with the general anesthesia followed by continuous electroacupuncture stimulation till the operation finished. The GA group was treated with simple general anesthesia. Changes of T cell subset, tumor necrosis factor-apha (TNF-alpha) and interleukin 6 (IL-6) were observed at time points of before anesthesia induction, 2 hours after operation, 1 and 3 days after operation and the occurrence of adverse reaction after operation was recorded. RESULTS: The percentages of CD3+ and CD8+ in both groups at 2 hours after operation were significant lower than that before anesthesia induction (all P < 0.05), and the percentage of CD4+ in the GA group decreased significantly at that time point (P < 0.05), while there was no significant difference in the ACGA group and CD4+/CD8+ increased significantly (P < 0.05). The level of TNF-alpha in the ACGA group decreased significantly at 3 days after operation as compared with that of before anesthesia induction (P < 0.05). There were significantly less cases nauseated after operation in the ACGA group than that in the GA group (P < 0.05). CONCLUSION: Acupuncture combine with general anesthesia has very little effect on immune function in patients of LC with less adverse reactions. PMID- 21644313 TI - [Effect of acupuncture on expression of matrix metalloproteinase and tissue inhibitor in cartilage of rats with knee osteoarthritis]. AB - OBJECTIVE: To explore the curative mechanism of acupuncture treatment on osteoarthritis (OA). METHODS: Forty cases of female SD rats were randomly divided into a normal group, a model group, an acupuncture group and a medication group, 10 cases in each group. OA animal model was established by using the method of heel tendon resection for unilateral hind limb. The acupuncture group was treated with electroacupuncture at "Xiqian"(ST 35) and "Housanli"(ST 36), and the medication group with inunction of Diclofenac cream, and the normal group and the model group without any treatment. The expression of matrix metalloproteinase-1, 3 (MMP-1, MMP-3) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the cartilage were observed by immunohistochemistry. RESULTS: There were significant differences among four groups. The expressions of MMP-1, MMP-3 and TIMP-1 in the model, acupuncture and medication groups were all significantly stronger than those in the normal group (all P < 0.01). The expressions of MMP-1 and MMP-3 in the acupuncture and medication groups were down regulated and TIMP-1 expression up-regulated with significant differences as compared with the model group (all P < 0.01), and the expressions of MMP-1 and MMP-3 in acupuncture group were significantly lower, while TIMP-1 expression significantly higher than that in the medication group (all P < 0.01). CONCLUSION: Acupuncture can down-regulate the expression of MMP-1 and MMP-3 and up-regulate the expression of TIMP1, which is superior to that of Diclofenac cream, showing that acupuncture has a certain protective effect on cartilage from OA. PMID- 21644314 TI - [Fifty-six cases of gastrocnemius muscle injury treated by superficial needling theray]. PMID- 21644315 TI - [Regulating effect of moxibustion pretreatment on Th1/Th2 imbalance of athletes during course of heavy load training]. AB - OBJECTIVE: To explore the effect of moxibustion pretreatment on the immune function of athletes during the courses of heavy load training. METHODS: Twelve male middle-distance race athletes were randomly divided into an observation group (n = 6) and a control group (n= 6). In observation group, mild moxibustion was applied at Guanyuan (CV 4), Zusanli (ST 36) or Guanyuan (CV 4), Sanyinjiao (SP 6) alternatively once daily, for 4 weeks. In control group, there was no any intervention. The expressions of gamma-interferon (IFN-gamma) and interleukin-4 (IL-4) were detected by realtime fluorescent quantitative PCR method before heavy load training, after 3 weeks of treatment, after 1 week adjusting training. RESULTS: (1) The ratios of Testosterone/Cortisol (T/C) of (0.017 +/- 0.003) in the control group and of (0.021 +/- 0. 007) in the observation group after heavy load training for three weeks were significant lower than 0.036 +/- 0.009 and 0.037 +/- 0.013 before heavy load training in the same group, respectively (both P < 0.01) (2) IFN-gamma mRNA of (2.76 +/- 1.28) and ratio of IFN-gamma mRNA/IL-4 mRNA of (3.28 +/- 1.76) in the control group after heavy load training for three weeks were significant higher than 1.75 +/- 0.44 and 1.52 +/- 0.76 before heavy load training, respectively (both P < 0.05), and ratio of IFN-gamma mRNA/IL-4 mRNA of (0.86 +/- 0.66) after adjusting training for 1 week reduced significantly as compared with that after heavy load training for three weeks (P < 0.05). (3) In the observation group, IFN-gamma mRNA, IL-4 mRNA and ratio of IFN-gamma mRNA/IL-4 mRNA increased firstly and then restore with the time, but these changes had no significant differences (all P > 0.05). CONCLUSION: Moxibustion pretreatment can prevent Th1/Th2 imbalance of male middle-distance race athlete during the course of heavy load training. PMID- 21644316 TI - [Xanthelasma treated by surround needling]. PMID- 21644317 TI - [Lumbar intervertebral disc protrusion treated with Santong tuina therapy: a multi-central randomized controlled trial]. AB - OBJECTIVE: To observe the difference in the efficacy on lumbar intervertebral disc protrusion (LIDP) between Santong tuina therapy and conventional tuina therapy. METHODS: Multi-central, random and controlled clinical trial was carried out. One hundred and twenty cases of LIDP were randomized into an observation group and a control group, 60 cases in each one. In observation group, Santong tuina therapy was used. In control group, conventional tuina therapy was adopted. The clinical efficacy was observed in two groups, and lumbar vertebral function was scaled before and after treatment in two groups. RESULTS: The total effective rate was 95.0% (57/60) in observation group and was 96.7% (58/60) in control group, without significant statistical difference in comparison (P > 0.05). The cured and markedly effective rate was 81.7% (49/60) in observation group and was 63.3% (38/60) in control group, indicating significant statistical difference in comparison (P < 0.05). The self-comparison of lumbar vertebral function before and after treatment presented statistical significance in either observation group (12.25 +/- 3.15, 25.56 +/- 5.27) or control group (13.32 +/- 3.26, 20.46 +/ 4.25, both P < 0.05); additionally, there was significant difference in the comparison between groups after treatment (P < 0.05). The therapy adopted in observation group improved lumbar vertebral function much significantly. CONCLUSION: In the treatment of LIDP, Santong tuina therapy achieves much better clinical efficacy as compared with conventional tuina therapy. PMID- 21644318 TI - [Drug dependence constipation in children treated by tuina]. PMID- 21644319 TI - [Clinical experiences of professor WU Bing-Huang on emergency treatment with acupressure]. AB - The key points of acupoint selection and manipulations of Professor WU Bing huang's experiences on emergency treatment with acupressure are introduced. It includes emergency treatment on coma (collapse, faint, faint at the sight of blood, faint during acupuncture, faint during moxibustion, shock, etc.), and pain, cough as well as asthma relieving with acupressure (include abdominal pain, vomiting, diarrhea, headache, toothache, dysmenorrhea, lumbago, neck stiffness after sleep, cough, asthma, etc.). At the same time, typical cases are given as examples. PMID- 21644320 TI - [Vicissitude and enlightenment of Bonghan theory]. AB - Dr. KIM Bonghan in North Korea in 1963 proclaimed that he discovered the physical structure of the acupoints and meridian systems which were corresponding to the Bonghan Corpuscles and Bonghan Ducts. The original discovery could not be replicated by other international researchers; therefore, it was neglected as a pseudoscience for years. This paper reviewed the relevant literatures which were published in the past 40 years, stated the findings published by Satoru Fujiwara A and Kwang-sup SOH and his colleagues. This paper also presented the perspective in meridian research after the Bonghan theory was proposed. PMID- 21644321 TI - [Experiences in prevention and treatment of human rabies with acupuncture and moxibustion in ancient time]. AB - By consulting the literatures relevant with the prevention and treatment of human rabies from Tang Dynasty to Qing Dynasty, it was discovered that rabies was caused by wind toxin in mad dog injuring human being. The pathogenesis of rabies was the invasion of dog toxin into the heart. The latent period was ranged from 7 to 100 days. Acupuncture-moxibustion, bloodletting, cupping and other therapies could be used in treatment. But of those different therapies, the various moxibustion methods achieved unique efficacy on the disease. PMID- 21644322 TI - [Study on the efficacy graded-disease-spectrum of acupuncture and moxibustion by the fuzzy comprehensive evaluation techniques: musculoskeletal and connective tissue diseases]. AB - The descriptive cross sectional surveys were carried out among the outstanding clinical physicians in the field of acupuncture and moxibustion by using the survey sampling. The fuzzy comprehensive evaluation techniques was applied to evaluate the opinions of clinical experts in the field of acupuncture and moxibustion in order to classify the spectrum of diseases related to the musculoskeletal and connective tissue. After the statistical analysis on sixty four diseases with 104 subtypes, the spectrum of diseases were determined as grade I spectrum of diseases with 13 subtypes of diseases, grade II spectrum of diseases with 51 subtypes of diseases, grade III spectrum of diseases with 10 subtypes of diseases, and grade IV spectrum of diseases with 30 subtypes of diseases. The outcomes reveal that acupuncture and moxibustion therapy have been applied widely for musculoskeletal diseases and connective tissue related diseases. PMID- 21644323 TI - [Discussion on "arrival of qi producing curative effects"]. PMID- 21644324 TI - [Essence of meridians and collaterals: circulatory conduction system of bio electricity of human]. AB - The running courses of twelve meridians are explained through the electrical properties of cell membrane, and the phenomenon related with meridians such as mechanism of acupuncture analgesia and acupuncture anesthesia, pause of the propagated sensation along channels, nature of propagated sensation and width of propagated sensation is expounded in this article. As a result, it is held that the meridian system, a circulatory conduction system of bio-electricity of human, is an independent system from the known nine large systems. PMID- 21644325 TI - [Perspectives of network thinking and network analysis applied on meridians and collaterals researches]. AB - The retrospective method of literatures was adopted and the epistemology of network thinking was integrated with the methodology of network analytic techniques so that the researches of meridian and collateral could be conducted in the new view of complex networks. The results indicate that meridian and collateral could be regarded as the complex networks composed of acupoints and meridians. The thinking and the methods of network analysis are ongoing to be applied to the paradigm of the analysis on meridian and collateral and it is expected that the paradigm could generate profound impacts on understanding the substance of meridian and collateral as well as on guiding clinical practice. PMID- 21644326 TI - [Fifty cases of alopecia treated by mild-warm moxibustion]. PMID- 21644327 TI - [Progress of treatment on oculomotor paralysis with electroacupuncture]. AB - Collecting information from the database of China National Knowledge Information (CNKI) and VIP database of Chinese Journals, an overall analysis and review on treatment of oculomotor paralysis with electroacupuncture in recently years are made in the paper. The electroacupuncture, which can provide constant stimulation and reinforce the effect of acupuncture, is the major therapy to treat oculomotor paralysis. However, the best stimulate threshold of electroacupuncture on oculorotary muscles has not been reported yet. The parameters of electroacupuncture adopted in clinic are not standardized, which lead to unfavorable therapeutic effect as well as unclear mechanism of treatment. Therefore, it is suggested that the study should focus on enhancing the research level and broadening researcher's mind to explore the best parameter for the electroacupuncture stimulation on the oculorotary muscles, to make clear the mechanism of treatment and to search for valuable observation indicators in the future. PMID- 21644328 TI - [Something is wrong with these hands. Did you notice?]. PMID- 21644329 TI - [Medical acute care in nursing homes. Family doctor can't be reached?]. PMID- 21644330 TI - [Eloquent aphasia patient]. PMID- 21644331 TI - [Guidelines. They were certainly not invented by family practitioners]. PMID- 21644332 TI - [Naturopathy consultation. Naturopathy for geriatric patients]. PMID- 21644333 TI - [Drugs in chronic heart failure. How many pills does the patient really need?]. PMID- 21644334 TI - [Too much alcohol and nicotine, unhealthy food, genital heat stress: fatherhood out of the question]. PMID- 21644335 TI - [The impact of body mass index on male infertility]. PMID- 21644336 TI - [Genital heat stress]. PMID- 21644337 TI - [Emergency checklist: frostbite]. PMID- 21644338 TI - [Practical management of acute and chronic cough]. PMID- 21644339 TI - [Baroflex stimulation: a novel treatment option for resistant hypertension]. PMID- 21644340 TI - [Blood pressure therapy - differential therapeutic solutions. Fixed combination improves prognosis of hypertensive patients]. PMID- 21644341 TI - [Cutting injuries in a 16-year-old. Is she a victim and/or perpetrator?]. PMID- 21644342 TI - [Naturopathy consultation. Vertigo]. PMID- 21644343 TI - [From beta blocker to relaxation exercises. Prevention of angina pectoris attacks]. PMID- 21644344 TI - [The price for complexity: hot spot shoulder joint]. PMID- 21644345 TI - [Impingement syndrome of the shoulder]. PMID- 21644346 TI - [Acromioclavicular joint osteoarthritis]. PMID- 21644347 TI - [Chondromatosis of the shoulder joint]. PMID- 21644348 TI - [Emergency checklist: tibial shaft fractures]. PMID- 21644349 TI - [Acute scrotal pain]. PMID- 21644350 TI - [Typical nail alterations in childhood]. PMID- 21644351 TI - ["Red eye" after thoracic drainage. Must it be treated?]. PMID- 21644352 TI - [When there is shortness of breath during the night - diagnosis and therapy of nocturnal asthma (interview by Dr. med. Brigitte Moreano)]. PMID- 21644353 TI - [The environment shapes the immune system. How dirt protects against allergies]. PMID- 21644354 TI - [Naturopathy consultation. Lumbar ischialgia]. PMID- 21644355 TI - [Current strategies against rhinosinusitis and otitis media]. PMID- 21644356 TI - [Acute rhinosinusitis]. PMID- 21644357 TI - [Diagnosis and treatment of otitis media]. PMID- 21644358 TI - [5-yr-old with borrelial lymphocytoma]. PMID- 21644359 TI - [Dental calculus--an increasing care problem in an aging society]. PMID- 21644360 TI - [Emergency checklist: spontaneous pneumothorax]. PMID- 21644361 TI - [Value of microalbuminuria in diagnostics of hypertension]. PMID- 21644362 TI - [Indication of EGFR kinase inhibitors should be refined]. AB - Even though lung cancer incidence began to decline in the majority of industrialized countries, is still belong to cancers with one of the highest incidence and mortality rates. In the Czech Republic, epidermal growth factor receptor (EGFR) kinase activity inhibitors erlotinib and gefitinib are approved for the use as the second- and third-line treatment of non-small-cell lung cancer. In a cohort of non-small-cell lung cancer patients, erlotinib administration led to tumour regression in less than 20% of patients. However, when used in patients with EGFR-activating mutations, e.g. L858R or delE746-A750, the response rate increased to 75-82% in several parallel clinical studies. Similarly, improved response rate was reported in patients bearing amplified wild type EGFR gene. In contrary, patients with T790M, D761Y, L747S, and T854A mutations (and some other rare abberations) were found to be resistant to treatment with small-molecule inhibitors targeting the active site of the kinase domain. These mutations do not change the EGFR affinity to gefitinib or erlotinib but the mutated receptor is able to bind ATP into its active site even in the presence of erlotinib or gefitinib, similar to a wild-type receptor without an inhibitor. Besides that, when the EGFR molecule bears both the activating (e.g. L858R) and resistance-inducing mutation (e.g. T790M), the tumour acquires resistance to both erlotinib and gefitinib treatment. Currently, research focuses on a development of new strategies that would allow treatment of patients bearing mutations inducing resistance to the small-molecule inhibitors targeted on the active site of EGFR kinase domain. Contrary to the current guidelines for Czech oncologists, identification of EGFR with any of the above mentioned resistance inducing somatic mutations should be considered as an explicit contraindication for non-small-cell cancer treatment using small-molecule EGFR kinase activity inhibitors erlotinib or gefitinib. This should also include patients in whom a resistance-inducing mutation is detected together with any of the activating mutations or deletions. PMID- 21644363 TI - [Adjuvant therapy in rectal cancer]. AB - BACKGROUNDS: Rectal cancer treatment has noticed several important changes in last years. TME, new radiotherapy techniques and new systemic agents have been introduced. Preoperative staging has improved. Nevertheless the neoadjuvant treatment approach is not unified all over the world. MATERIAL AND METHODS: Using the following American and European guidelines we found primary citations and compared their results. CONCLUSION: Currently, there are substantial differences in neoadjuvant dose of radiotherapy between European and American guidelines. Short neoadjuvant course of radiotherapy seems to be appropriate in patients with smaller primary operable rectal cancer.This approach is safe if chosen, not oppressing stage III patients by protracted oncological treatment including chemotherapy. PMID- 21644364 TI - [Avastin in the treatment of breast cancer]. AB - Over the last decades, various new agents have been developed for the treatment of metastatic breast cancer and overall survival of these patients has increased. The role of chemotherapy in the treatment of metastatic breast cancer is well established. Bevacizumab is a potent antiangiogenic agent active in many solid tumors. Three randomised clinical trials (E2100, AVADO and RIBBON I) proved the benefit of chemotherapy, especially a combination of taxanes and bevacizumab, as the first line treatment of metastatic breast cancer. The combination improved patient progression-free survival in all trials with no impact on the known toxic effects of taxanes. This may be a potent treatment option particularly for patients with triple negative breast cancer, and a potentially less toxic alternative to combination chemotherapy. PMID- 21644365 TI - [Overview of potential oncomarkers for detection of early stages of ovarian cancer]. AB - The causes of ovarian cancer have not been fully elucidated yet but genetic predisposition is found in approximately 10% of patients. When the disease is detected at an early stage, up to 90% of patients have a hope of recovery. However, no preventive measures or precise screening tests to detect early stages of this disease are known yet. Standard tumor markers (CA125) are usually investigated in women with an increased risk. Nevertheless, due to low sensitivity and specificity during the first stage of the cancer, CA125 determination showed a very low efficacy (less than 26%). There has been a considerable progress over the recent years in understanding the molecular mechanisms leading to tumor formation and metastasis. Gradually, 46 genes were identified, initially named tumor endothelial markers (TEM), the expression of which is increased in tumors compared to normal endothelial cells. Death receptor 6 (DR6) and glycoprotein M6B (GPM6B), both detectable from patients serum, are among the most promising candidates for a marker of an early stage of ovarian cancer. This review aims to clearly describe potential as well as clinically used tumor markers useful in an early detection of ovarian cancer. Search for new markers, characterized by increased expression in patients'blood is a highly topical issue. PMID- 21644366 TI - [Multimodal treatment of glioblastoma multiforme: results of 86 consecutive patients diagnosed in period 2003-2009]. AB - BACKGROUNDS: Glioblastoma multiforme is the most common malignant primary tumor of the brain in adults. Standard therapy consists in maximal surgical resection and adjuvant concurrent chemoradiotherapy and adjuvant therapy with temozolomid. This approach improves survival in comparison with postsurgical radiotherapy alone. PATIENTS AND METHODS: Consecutive patients with histologically confirmed glioblastoma multiforme in the period from January 2003 to December 2009 underwent postoperative radiotherapy (1.8-2.0 Gy/d, total of 60 Gy) plus concurrent daily chemotherapy (temozolomide 75 mg/m2/d), followed by 6 cycles of temozolomide (150 to 200 mg/m2 for 5 days, every 28 days) and were analyzed retrospectively. The primary end point was to describe the correlation between known clinical factors, treatment and progression free survival (PFS) and overall survival (OS). We assessed the toxicity and safety of the chemoradiotherapy. RESULTS: Eighty-six patients (median age, 56 years; 60% male) were included. Most of them (> 80%) were of performance status (PS) 0-1 at the beginning of chemoradiotherapy. Total macroscopic resection was performed in 20% of the patients, subtotal in 65%, partial in 9%, and just biopsy in 6%. Median PFS was 7.0 months (2.0-35.5), median OS was 13.0 months (2.5-70). Postoperative performance status (PS), the extent of resection, and administration of planned treatment without reduction had statistically significant influences on PFS and OS. Median PFS and OS were 22.0, 7.0 and 6.0 months for PFS (p = 0.0018) in patients with PS O, 1 and 2 respectively and 32.0, 13.0 and 9.0 months for OS (p = 0.0023). Patients with total removal of tumor had longer PFS (14.0 vs 6.0 months, HR = 0.5688; p = 0.0301) and OS (23.0 vs 12.0 months, HR 0.4977; p = 0.0093), as did patients without dose reduction of radiotherapy and/or chemotherapy. Patients with radiotherapy dose of over 54 Gy had PFS 8.0 vs 3.0 months (HR = 0.3313; p = 0.0001) and OS 15.0 vs 5.0 months (HR = 0.1730; p < 0.0001). Similarly, treatment with concurrent chemotherapy for more than 40 days was also important: PFS 8.0 vs 5.0 months (HR = 0.5300; p = 0.0023) and OS 17.0 vs 9.5 months (HR = 0.5943; p = 0.0175). Age, gender and position of tumor had no significant influence. Treatment-related hematology toxicity grades 3 and 4 occurred relatively often: thrombocytopenia (9%), leukopenia (6%), neutropenia (6%) and lymphopenia (25%). Thrombo-embolic events were dominant in non hematology toxicity. Serious toxicity occurred mainly in the subgroup of patients with PS 2. Treatment of progression was useful in selected patients. Second surgery was of the most benefit (OS 24.0 vs 12.5 months, HR = 0.5325; p = 0.0111). CONCLUSION: Postoperative performance status, extent of resection, successful administration of the majority of planned concurrent chemoradiotherapy and possibility of surgical treatment at the time of recurrence correlate with better prognosis for our patients with glioblastoma. Our experience indicates that performance status should be the main factor in decisions about treatment intensity. Treatment of malignant glioma requires a multidisciplinary team. PMID- 21644367 TI - [Treatment of patients with relapsed/refractory Hodgkin lymphoma]. AB - BACKGROUNDS: This retrospective study evaluated treatment outcomes in patients undergoing autologous stem cell transplantation (ASCT) for relapsed/refractory Hodgkin lymphoma (HL). PATIENTS AND METHODS: Overall, 194 HL patients treated with ASCT between 2000 and 2009 were analyzed. Survival was calculated using Kaplan-Meier method and differences in survival between subgroups with log-rank test. RESULTS: Best responses observed after ASCT: 124 complete and 35 partial remissions, 2 patients with stable disease and 33 relapses/progressions. During a median follow-up of 44 months, seventy patients after ASCT progressed/relapsed. Thirty-seven patients received salvage chemotherapy only with or without radiotherapy, 25 underwent allogeneic stem cell transplantation (SCT), 4 the second ASCT and 4 refused treatment. 5-year overall survival after ASCT was 71% and progression-free survival 54%. Median survival of the 70 patients relapsing after ASCT was 16.9 months. Median survival in patients after allogeneic SCT was 31.8 months and 12.4 months in patients treated with other modalities (p = 0.21). Overall mortality was 26.3% (51/194 patients): 13.4% progressions/relapses of HL and 12.9% non-relapse mortality. CONCLUSION: Efficacy of ASCT was confirmed in 54% progression-free survivors. Median survival after ASCT failure is relatively short. There is a slightly longer overall survival after allogeneic SCT, although not statistically significant when compared to other approaches. PMID- 21644368 TI - [Metastatic choriocarcinoma in 26-year-old woman--case report]. AB - Gestational trophoblastic tumors are a group of diseases spanning from a benign hydatidiform mole, through an invasive mole to a highly malignant form of choriocarcinoma. Choriocarcinoma is a rare but the most malignant and aggressive neoplasm of all the gestational trophoblastic diseases. It grows rapidly and metastasizes into the lungs, liver, and, less frequently, to the brain. The author presents a case of a 26-year-old woman who was, based on previous investigations, referred for a CT examination for suspected brain tumour. On the basis of medical history provided by the patient's husband, high beta-hCG level and a gynaecological examination, the patient was diagnosed with choriocarcinoma metastasizing into the brain, kidneys and adrenal glands. PMID- 21644369 TI - [Therapeutic equivalence and effectiveness of Binocrit (HX575) in patients with anemia caused by chemotherapy]. PMID- 21644370 TI - [Comments on the TNM classification of malignant tumours--7th edition]. PMID- 21644371 TI - [Course and conclusions of the interdisciplinary meeting "Winter GLIO TRACK Meeting" 2011]. PMID- 21644372 TI - [Problem drug resistance]. PMID- 21644373 TI - [Vernakalant--a new drug for pharmacological conversion of atrial fibrillation]. AB - Vernakalant is a new antiarrhythmic drug which is used for rapid conversion of atrial fibrillation. The drug is a relatively atrium-selective K' and Na+ channel blocker, prolonging the atrial refractory period. Clinical trials showed conversion rates of about 50% which is comparable to other antiarrhythmic drugs. Further trials will show if vernakalant is less pro-arrythmogenic than its competitors. PMID- 21644374 TI - [Renaissance of bed bugs]. PMID- 21644375 TI - [S3-guideline for uncomplicated urinary tract infections - treatment guidelines compliance]. PMID- 21644376 TI - [National care guideline for back pain]. PMID- 21644377 TI - [Is the clinical research crisis a measurement error?]. PMID- 21644378 TI - [Leading anesthesiologist accused of scientific misconduct. We are not changing our protocols--yet]. PMID- 21644379 TI - [The Swedish palliative registry has scientific shortages. But it's suitable for own quality work]. PMID- 21644380 TI - [Big gaps in documentation of end-of-life care. Incomplete medical records reported to the Swedish Registry of Palliative Care]. PMID- 21644381 TI - [Incidents involving concentrated potassium and sodium solutions. Analysis and lessons learned from reported cases]. PMID- 21644382 TI - [Cephenemyia ulrichii larvae in the eye--unusual but serious problem. Cases of human ophthalmomyiasis from Dalecarlia and southeastern Finland]. PMID- 21644383 TI - [Reverse implementation of "the Gold-mine"]. PMID- 21644384 TI - [Adverse effect of Sifrol or not?]. PMID- 21644385 TI - [Otitis studies show that Swedish recommendations hold]. PMID- 21644386 TI - Lwow School of Dermatology between World War I and II. PMID- 21644388 TI - Medicaid program; payment adjustment for provider-preventable conditions including health care-acquired conditions. Final rule. AB - This final rule will implement section 2702 of the Patient Protection and Affordable Care Act which directs the Secretary of Health and Human Services to issue Medicaid regulations effective as of July 1, 2011 prohibiting Federal payments to States under section 1903 of the Social Security Act for any amounts expended for providing medical assistance for health care-acquired conditions specified in the regulation. It will also authorize States to identify other provider-preventable conditions for which Medicaid payment will be prohibited. PMID- 21644389 TI - Can newer diagnostic microbiological assays guide early tuberculosis management? PMID- 21644387 TI - [Natural killer cells and cancer. Regulation by the killer cell Ig-like receptors (KIR)]. PMID- 21644390 TI - Prevalence of XDR TB cases--a retrospective study from a tertiary care TB hospital. AB - BACKGROUND: The emergence of XDR -TB strains is a major roadblock in the successful implementation of TB control programmes. This further leads to high morbidity and mortality, especially in immuno-compromised patients. Identification and observation of resistance patterns of XDR-TB strains may help clinicians manage MDR-TB cases, the treatment line of which is expensive, time taking and involves intake of toxic drugs with many side-effects. Our study is aimed to find out the prevalence of XDR-TB among the MDR-TB strains isolated in a tertiary care hospital. MATERIAL & METHODS: The study population consisted of 223 patients of tuberculosis who were culture positive and Mycobacterium tuberculosis was resistant to Rifampicin and Isoniazid during January 2007 to December 2009. Each patient had submitted two sputum samples i.e. spot and morning. The identified Mycobacterium tuberculosis complex was subjected to drug sensitivity testing by first and second line drugs by proportion and absolute concentration methods as per standard procedure. RESULTS: The results showed that 20.17% strains (45/223) were XDR-TB strains. Most of these strains showed resistance to four drug combination viz. KM, ETH, OFX & PAS (5.82%), KM & OFX (3.13%), OFX, KM and ETH (1.79%), 1.34% strains showed resistance to all the drugs i.e. pan resistance and other combinations in the remaining strains. Nearly 80% of the XDR TB strains showed resistance to three or more drugs combination pattern. CONCLUSION: The multidrug resistant TB cases need urgent and timely sensitivity report for second line ATT drugs to help clinicians start proper drug combinations to treat MDR-TB patients. PMID- 21644391 TI - Incremental yield in sputum smear positivity by examining a second early morning sputum specimen in follow-up patients on DOTS: 7 year analysis of RNTCP laboratory register. AB - BACKGROUND: Patients receiving DOTS undergo periodic follow-up sputum examination, which aids in monitoring response to treatment. Continued or new smear positivity at follow up examination entails extension of intensive phase or change in treatment category and the need for culture and drug susceptibility test. SETTING: Tuberculosis microscopy centre at a tertiary care teaching hospital, Mumbai, India. OBJECTIVE: To determine the incremental yield in sputum smear positivity by examining a second early morning sputum specimen in follow-up patients on DOTS. DESIGN: Retrospective analysis of follow up sputum microscopy results recorded in tuberculosis laboratory register for the period 2002-2008. RESULTS: During the study period, 5015 follow-up patients submitted two early morning sputum specimens, of which 501(9.99%) patients were detected to be smear positive. Out of smear positive patients under study, 324 patients had both specimens positive, 79 patients had only first specimen positive and 98 patients had only second specimen positive. The incremental yield was 1.95% of total and 19.5% of smear positives. CONCLUSION: Discordant smears were present in nearly a third of patients detected smear positive during follow-up. More than half of these patients were detected only by examining second specimen. The incremental yield by examining the second early morning specimen was 1.95% of total and 19.5% of smear positive specimens. It is important to detect each possible smear positive follow-up patient as they are likely to benefit from altered treatment. The inclusion of a second early morning sputum specimen examination is essential to maximize their detection. PMID- 21644392 TI - Acute suppurative presentation of osteoarticular tuberculosis in children. AB - INTRODUCTION: Osteoarticular tuberculosis is known for its atypical presentations. The acute presentation of osteoarticular tuberculosis although mentioned by many observers is seldom seen in practice. We report the varied presentations of acute suppurative presentation of osteoarticular tuberculosis in pediatric age group. METHODS: Our study retrospectively assessed 10 children with osteoarticular tuberculosis who had acute presentation with short history of a few days and signs of acute inflammation. RESULTS: The age range varied from 3-12 years. The duration of symptoms averaged 14.7 days (range, 4-28 days). Three patients were afebrile at the time of presentation. The diagnosis of tubercular aetiology was made only retrospectively in all the cases. All, except one, were subjected to Fine Needle Aspiration Cytology (FNAC)/ trochar bone biopsy. Diagnosis was based on smear positive for acid fast bacilli (n = 3), histopathology (n = 5), and on clinicoradiological findings in two cases. The acute exudative pattern was seen in seven and gravity assisted tracking and accumulation of abscess (dependent) in three patients. In eight patients, the FNAC/ trochar biopsy decompressed the lesion initially. Incision and drainage were performed on one case of osteoarticular tuberculosis with clinical presentation of acute pyogenic infection. Another patient of acute exudative pattern was subjected to drainage and debridement in view of impending burst. Both exudative and dependent forms of suppurative presentations of osteoarticular tuberculosis responded well to standard antitubercular chemotherapy. The abscesses resolved within a period of 6-12 weeks. CONCLUSION: The acute suppurative presentation is a rare and atypical form of osteoarticular tuberculosis. It has close resemblance to acute pyogenic infections or septic arthritis and pose significant diagnostic dilemma for the unwary. A vigilant and methodical approach is the key for managing acute suppurative tubercular presentations. PMID- 21644393 TI - Comparison of Ziehl Neelsen & Auramine O staining methods on direct and concentrated smears in clinical specimens. AB - BACKGROUND: In developing countries like ours with a large number of tuberculosis (TB) cases and limited resources, the diagnosis of TB relies primarily on smear microscopy for Acid Fast Bacilli (AFB) but its sensitivity is limited in paucibacillary cases. AIM: To evaluate the increase in efficacy of smear microscopy when smears are prepared from clinical samples after concentration by Petroff's method and stained by Auramine O (AO) fluorescent dye as against Ziehl Neelsen (ZN) staining of similar taking culture as the gold standard. METHODS: Smears were prepared from 393 clinical samples both by direct and after Petroff's concentration and examined by fluorescent microscopy and Ziehl Neelsen method .The concentrated material was also cultured on Lowenstein Jensen media and the results of the two microscopy methods were compared with the culture results taken as the gold standard. RESULTS: Mycobacterial growth was detected in 137 (35.77%) specimens, out of which three were non-tubercular mycobacteria. Using culture as the reference method, the sensitivity of direct staining was 55.55% for ZN and 71.85% for AO. Direct fluorescent microscopy detected 9.29% paucibacillary sputum samples that were missed on ZN staining. On concentration, the sensitivity increased by 6.67% for ZN and 11.11% for AO. The sensitivity of AFB smear microscopy increased by 27.41% and was statistically significant (p = < .001) when both methods were combined. The specificity was 99.19% for both ZN and AO. CONCLUSION: Fluorescent microscopy has higher sensitivity and comparable specificity which is further enhanced by concentration. Now with the advent of newer inexpensive Light Emitting Diode (LED) based fluorescent microscopes (FM), which are easier to use, fluorescent microscopy can be widely used even in peripheral laboratories where culture facilities are not available. PMID- 21644394 TI - Does CSF composition predict shunt malfunction in tuberculous meningitis? AB - BACKGROUND: Hydrocephalus secondary to tuberculous meningitis (TBM) continues to be a challenging condition to treat for neurosurgeons in developing countries. Shunt complications are reportedly more frequent in patients undergoing ventriculo-peritoneal shunt in patients with TBM than in those undergoing shunt surgeries for other causes. AIM: The aim of this study was to evaluate the relationship of cerebrospinal fluid (CSF) composition on shunt malfunction. METHODS: We compared the CSF composition of 53 patients who had shunt malfunction during a five year period with that of 137 matched controls. RESULTS: Patients who had shunt malfunction had a significantly higher concentration of CSF protein. The CSF cellularity and glucose concentration did not have any significant bearing in predicting shunt malfunction. Patients with CSF protein concentration of more than 200 mg/dL had a four times higher risk of having shunt malfunction than those with a concentration of less than 100 mg/dL. Patients with CSF protein in the 100-200 mg/dL range represent an intermediate zone. CONCLUSION: To conclude, patients with CSF protein concentration of more than 200 mg/dL have a significantly higher risk of shunt malfunction and hence have to be followed up closely. PMID- 21644395 TI - A case of adrenal tuberculosis with pulmonary tuberculosis. PMID- 21644396 TI - Pulmonary embolism in cases of pulmonary tuberculosis: a unique entity. AB - Pulmonary tuberculosis is very prevalent in developing countries but its thrombogenic potential is a new entity. There are reports stating the relation of Deep Vein Thrombosis (DVT) with severe forms of tuberculosis but no literature is available for correlation of pulmonary tuberculosis and pulmonary embolism. We are presenting series of five patients with different forms of tuberculosis presenting with pulmonary embolism having no risk factor for hypercoagulability. Also, serum protein C, protein S, antithrombin and factor V levels were normal in all. We are highlighting an unreported phenomenon so that high suspicion, adequate prophylaxis and prompt management of pulmonary embolism can play a vital role in the survival of this subset of patients. PMID- 21644397 TI - Blastoschizomyces capitatus pneumonia in an immuno-competent female. AB - Pulmonary infection due to Blastoschizomyces capitatus is less common. It is an emerging fungal pathogen. We describe a case of Blastoschizomyces capitatus pneumonia in an otherwise healthy female and review the clinical presentation, microbiological characteristics, and treatment for B. capitatus infection. PMID- 21644398 TI - Status report on RNTCP. PMID- 21644399 TI - Anti-Mullerian hormone in women with polycystic ovary syndrome. AB - Anti-Mullerian hormone (AMH) is a glycoprotein produced in the granulosa cells of the ovary. It is involved in the regulation of follicular growth and development. AMH serum and follicular fluid concentration is increased in women with polycystic ovary syndrome (PCOS), which correlates with the extent of ovarian dysfunction and clinical manifestation of the syndrome. It is unclear whether the higher AMH levels in PCOS are due to the higher number of preantral follicles or result from a specific disorder in the synthesis of AMH causing follicular arrest in PCOS. AMH determination has high specificity and sensitivity as a diagnostic marker for PCOS. The AMH level can also be used to predict the effect of treatment in PCOS women, the higher values implying more difficulties in the therapeutic management of the disease. PMID- 21644400 TI - Relationship of physical activity and eating behaviour with obesity and type 2 diabetes mellitus: Sofia Lifestyle (SLS) study. AB - OBJECTIVE: Physical inactivity and excessive food consumption play a major role in the etiology of obesity and type 2 diabetes mellitus (T2DM). The aim of the present study was to investigate the relationship of physical activity (PA) and eating behaviour with obesity and T2DM in citizens of Sofia, Bulgaria. PATIENTS AND METHODS: A total of 511 randomly chosen participants completed a validated questionnaire concerning age, body height and weight, medical history, and motivation for a lifestyle change. The Baecke PA questionnaire and the Three Factor Eating Questionnaire were also completed. RESULTS: Body mass index (BMI) was significantly higher among subjects in the lowest compared with the middle (P = 0.002) and with the highest tertile of leisure time PA (P < 0.001), also between the lowest and the highest tertile group of sport PA (P = 0.001). BMI differed significantly in the highest vs. the middle (P = 0.04), as well as vs. the lowest (P= 0.017) tertile of uncontrolled eating behaviour. The prevalence of T2DM was significantly greater in the lowest vs. the middle (P = 0.027) and the highest (P = 0.02) tertile of leisure time PA. In a multiple regression analysis both leisure time PA and uncontrolled eating were independently associated with BMI (beta = -0.13, 95% CI -1.83 to -0.11, P = 0.03 and beta = 0.32, 95% CI 0.23 to 0.44, P < 0.001). CONCLUSIONS: We found a strong inverse relationship between the level of PA during leisure time (including sport), BMI, and the prevalence of T2DM. Uncontrolled eating behaviour was also found to have a significant effect on BMI. PMID- 21644401 TI - Predictive factors for high brain (B-type) natriuretic peptide at discharge in properly treated heart failure patients. AB - AIM: To study differences and prognostic effect of some factors on brain (B-type) natriuretic peptide (BNP) levels at discharge of patients with chronic heart failure (CHF) treated in accord with current treatment guidelines. PATIENTS AND METHODS: Eighty-five consecutive patients hospitalized for CHF were recruited into the study. A standardized study protocol was used for them including collection of blood samples for measurement of electrolytes, creatinine at baseline and BNP at discharge. High BNP levels were determined at values above the threshold value for the highest BNP quartile (> or = 463 pg/ml). Linear regression analyses were performed using the SPSS 16.0. RESULTS: High BNP levels at discharge were measured in 21 (24.7%) of the patients. Patients with persisting high BNP levels, despite the administered correct therapy, had significantly more frequently worse clinical and instrumental characteristics: pulmonary congestion (76.2% vs. 40.6%), IV NYHA functional class (23.8% vs. 4.7%), atrial fibrillation (AF) (71.4% vs. 35.9%) and ischemic etiology of HF (47.6% vs. 15.6%). Multivariate linear regression analysis (F = 7.1, p < 0.001) identified systolic blood pressure (SBP), AF and instrumental data for pulmonary congestion as significant and independent predictors of high BNP at discharge. CONCLUSIONS: There were statistically significant differences in the distribution and prognostic effect of the studied factors across the BNP levels. CHF patients with hypotension, AF and instrumental evidence for pulmonary congestion at admission are at higher risk of high BNP levels at discharge and require special care and clinical approach. PMID- 21644402 TI - Plasma asymmetric dimethylarginine levels in healthy people. AB - Asymmetric dimethylarginine (ADMA) is an endogenous competitive inhibitor of the endothelial nitric oxide synthase (eNOs). ADMA is believed to be implicated in angiogenesis because it regulates the nitric oxide biosynthesis; any pathological abnormalities in ADMA play a crucial role in the pathogenesis and progression of atherosclerosis. The AIM of the present study was to determine the reference range for plasma concentration of ADMA in a sample of Bulgarian population. PATIENTS AND METHODS: To establish the reference interval of ADMA plasma levels and study the impact of sex and age we recruited 150 healthy subjects of Bulgarian nationality aged between 18 and 65 years. The selection criteria for the reference group were made to comply with the generally approved recommendations of the International Federation of Clinical Chemistry (IFCC). Plasma concentrations of ADMA were determined using ELISA assay (DLD, Diagnostics, Hamburg, Germany). RESULTS: The reference ranges for ADMA, given as 95% of the measured values, were from 0.22 to 0.69 micromol/1. We found no sex related differences in ADMA concentration (P > 0.05), which obviates the need for separate reference intervals for men and women. Single-factor dispersion analysis found no age dependency ofADMA (P > 0.05, F = 1.038) in the studied reference group in the age range 18-65 which makes unnecessary establishment of reference intervals for lower age ranges in this age group. CONCLUSION: The reference values for ADMA plasma concentrations calculated according to the type of distribution of results can be used as baseline criteria in clinical laboratory studies and for clinical purposes. PMID- 21644403 TI - Ultrasound tomography in diagnosing cystic pancreatic neoplasms. AB - The clinical, radiologic, and pathologic features of cystic pancreatic lesions are well known, and yet preoperative diagnosis is often difficult to make. AIM: To present the role of ultrasound study in conventional and interventional diagnostics of cystic pancreatic neoplasms. PATIENTS AND METHODS: 65 patients (17 men, 48 women) with cystic pancreatic lesions were examined by ultrasound and computer tomography and investigated using a number of diagnostic interventional procedures over three years. We used Siemens-Adara ultrasound system, GE Sytec 3000i CT equipment and a set of biopsy needles and catheters. RESULTS: We performed ultrasound studies for all 65 patients to determine the structural characteristics of the cystic lesions. Lesions varied in size from 15 to 130 mm in diameter. In 50 patients we used US control to perform the invasive procedures for cytological and histological examination. In 9 cases the invasive procedures were repeated. In 15 cases we preferred to use CT guidance. We were able to detect 46 pseudo cysts, 9 mucinous neoplasms, 6 serous cystadenomas, 3 intraductal papillary mucinous neoplasms and 2 lymphoepithelial cysts. CONCLUSIONS: Ultrasound tomography is an imaging method of sufficient informative value with respect to the structural characteristics of cystic pancreatic neoplasms. US-guided invasive manipulations of cystic pancreatic lesions are easy to perform, quick and effective diagnostic methods. In some cases punctures and biopsies under CT control appear to be the imaging modality of choice. PMID- 21644404 TI - Disability-related injuries in athletes with disabilities. AB - OBJECTIVES: Athletes with disabilities are highly skilled. Sports-related injuries and disorders interfere with their efforts. Several aspects of these injuries have been studied in previous studies. The aim of this study was to correlate the types of injuries with the disability group in athletes with physical disabilities. METHODS: One hundred and thirty nine elite athletes with physical disabilities completed a questionnaire about sports-related injuries that resulted in at least one day off from training or competition. RESULTS: All disability groups show soft tissue injuries in high percentages. Cerebral palsy (CP) athletes reported soft tissue injuries (P < 0.01) and lacerations (P < 0.001) in higher percentage than Other Disabled Athletes (ODA) and Spinal Cord Injured (SCI) athletes. Spinal cord injured athletes sustained fractures (P < 0.05) and blisters (P < 0.05) in higher percentages than the other groups. No differences were found between the studied groups for contusions, low back pain, ruptures, thermoregulation disorders, urinary tract infections, pressure sores and pneumonias. CONCLUSION: CP athletes sustained soft tissue injuries and lacerations more than other disability groups did because moving and walking patterns of this population add risk factors for such injuries. Fractures and blisters occur more frequently to SCI athletes because they participate in higher percentage in wheelchair basketball which is high risky sport. PMID- 21644405 TI - Our experience in the diagnostics and therapy of patients with solitary peripheral lung tumours. AB - Patients presenting with a solitary pulmonary nodule are often a serious diagnostic-therapeutic problem. The AIM of the present report is to share our experience with VATS in the diagnostics and therapy of patients with solitary pulmonary nodules. PATIENTS AND METHODS: Between 2005 and 2009 we performed primary videothoracoscopic tumor resection in 125 patients with peripheral lung tumours. RESULTS: In 93 (74.4%) of the cases complete successful thoracoscopic resection of the tumor and subsequent intraoperative frozen section histology was accomplished. In the remaining 32 cases (25.6%) various reasons demanded conversion to thoracotomie with wedge resection of the nodule. 54.2% of the removed tumours were malignant and 45.8% - benign. In 18 of the patients the intraoperative frozen section histology diagnosis was primary bronchial carcinoma which required to do oncologic resection of the tumor. In 10 patients the intraoperative frozen section histology was unable to differentiate between pulmonary metastasis and primary bronchial carcinoma, and the oncologic resection was performed in another surgical session after the definitive pathomorphologic diagnosis had been available. In 28 of the cases pulmonary metastases of a primary extrapulmonary tumor were removed. CONCLUSIONS: Despite improvement in the diagnostic techniques, only the video-thoracoscopic removal of the lung nodule and its subsequent histological examination enables the definitive diagnosis, solving definitively the problem with benign lesions and indicating appropriate treatment of the malignant pulmonary lesions. PMID- 21644406 TI - Effects of ketamine on memory and nociception in rats. AB - BACKGROUND: Ketamine is intravenous anaesthetic with NMDA-glutamate receptors mechanism of action. MATERIAL AND METHODS: Male Wistar rats were treated with saline (group A) or 10, 15 or 20 mg/kg of ketamine (groups B, C and D, respectively). For active avoidance test an automatic reflex conditioner was used. The observed variables were number of avoidances, escapes and intertrial crossings. Step-through and step-down passive avoidance tests were done with learning and memory retention test. Criteria for step-through test were latency of reactions of 180 sec in the light chamber. Criteria for step-down test were latency of reaction of 60 sec on the platform. The hot-plate test evaluates the reaction time of the rats dropped on a heated surface. The analgesy-meter test exerts a force increased at constant rate. RESULTS: In active avoidance test the controls increased the number of avoidances during learning and memory tests. Ketamine in all doses used increased the number of avoidances during learning and in memory test. Controls did not change the number of escapes, but the ketamine treated animals decreased it. The number of intertrial crossings was not changed by controls or ketamine-treated rats during learning and memory tests. In passive avoidance tests the controls and ketamine-treated rats increased the latency time during learning and memory retention tests. In hot-plate analgesic test and in analgesy-meter test the controls and ketamine-treated rats did not change the latency of reaction. CONCLUSION: The results suggest that ketamine improves learning and memory processes and has no analgesic effect in the doses applied. PMID- 21644407 TI - Resolution of experimental pneumothorax by room air. AB - Observation alone is advised only for primary spontaneous pneumothoraces with less than 20 percent of the lungs collapsed. In such cases, it is the resorption capabilities of the visceral pleura that are solely relied upon. The AIM of the present experimental study was to demonstrate the capabilities of the pleura for pneumothorax resolution by room air. MATERIAL AND METHODS: The study was conducted with six laboratory animals (New Zealand white rabbits, weighing 2.5 3.5 kg), in which right total pneumothorax was induced. Conventional chest X-rays at intervals of 2 days were used to monitor the changes in the size of the pneumothorax until its final resolution. The size of the pneumothorax was estimated by the Light index. RESULTS: Total resolution of the pneumothorax occurred within 12 days after it was induced. The average rate of pneumothorax resolution was 6.63% (SEM +/- 0.20)/daily. CONCLUSION: The study imitates initial clinical observation in pneumothorax. Despite the fact that the study demonstrates resolution of total pneumothorax (100%), observation only is inappropriate as a treatment modality in patients with more than 20% collapse of the lung. PMID- 21644408 TI - Delayed posttraumatic unilateral occipital epidural hygroma in early childhood. AB - An extremely rare case of posttraumatic epidural hygroma in the left occipital supratentorial and infratentorial region is reported. A year and five months old child was admitted to the Clinic of Neurosurgery with sustained occipital head injury. She presented with drowsiness and vomiting due to intracranial hypertension. Initial computed tomography scan revealed left-sided fracture of the squamous part of the occipital bone without associated traumatic changes to the brain. A second spiral computed tomography scan was obtained two days later because of persisting symptoms of increased intracranial pressure. It demonstrated a newly formed left-sided epidural hygroma adjacent to the skull fracture in the left supratentorial and infratentorial occipital region. The case is discussed with emphasis on the mechanism of formation of epidural hygroma and an attempt has been made to outline the major predisposing factors leading to the development of this traumatic disease. Necessity for computed tomography follow up is pointed out in order to diagnose delayed posttraumatic hygromas. The recommended surgical approach should include craniotomy centered at the site of the epidural hygroma and obligatory dural elevation by means of traction sutures to eliminate the posttraumatic epidural cavity. PMID- 21644409 TI - Issues around HIV-1 diagnosis. PMID- 21644410 TI - Factors associated with delayed diagnosis of HIV infection in Mukono District, Uganda. AB - OBJECTIVE: To identify factors associated with delayed diagnosis of HIV infection. DESIGN: Cross sectional study. SETTING: Mukono district, Uganda. SUBJECTS: Newly diagnosed HIV positive clients at ten HIV testing centres. Late testers were HIV positive clients who had AIDS according to World Health Organisation AIDS case definition. Early-testers did not have AIDS at diagnosis. MAIN OUTCOME MEASURES: Late testers were compared to early testers on socio demographic characteristics, sexual behaviour, access of testing services, knowledge of care and support services for HIV/AIDS and attitude towards knowing HIV-status. RESULTS: Delayed diagnosis of HIV infection was independently associated with being over 25 years (adjusted odds ratio (AOR), 4.3; Confidence Interval (CI) 1.7-11.1), not being married (AOR, 2.4; CI 1.3-4.4), having no knowledge of testing services (AOR, 2.4; CI, 1.2 4.7), spending at least one hour travelling to a testing centre (AOR, 2.1; CI, 1.0-4.2), paying for HIV testing (AOR, 3.4; CI, 1.7-6.9) having had an illness two months before testing AOR 9.49; CI, 4.84-18.64) and having negative beliefs towards knowing one's HIV sero status (AOR, 5.7; CI, 1.0-30.8). CONCLUSION: Factors associated with delayed diagnosis of HIV infection in Mukono District of Uganda are; age over 25 years, not being married, having no knowledge of testing services, paying for HIV testing, travelling for at least one hour to a testing centre and having negative beliefs towards knowing one's HIV status. In order to increase access to testing there is need to offer free HIV testing and health education targeting people above 25 years and those not married. PMID- 21644411 TI - Diagnosis of dual human immunodeficiency virus types 1 and 2 infections in a resource-limited setting. AB - BACKGROUND: The presence of dual HIV-1/HIV-2 infection in Ghana and the different drug requirements for the treatment of HIV-1 and HIV-2 presents difficulties for the treatment of dual infections with both viruses. OBJECTIVES: To determine the prevalence of the dual sero-positive profile in treatment naive patients at a principal ART Clinic in Accra, Ghana and to investigate if rapid screening assays could be useful for diagnosis. DESIGN: A cross-sectional study. SETTING: A principal antiretroviral treatment centre in Accra, Ghana. SUBJECTS: Three hundred and twenty eight antiretroviral treatment naive patients. RESULTS: A total of 12 (3.7%) of patients seen were dual seropositive. There was a slight tendency of dual seropositive females being older than their HIV-1 counterparts (p = 0.088, CI = -10.833 to 0.753). Eight of the 12 of the dual seropositives were reactive for Genie II and were considered as possibly infected with both HIV I and HIV-2. Seven (87.5%) of Genie II dual seropositives had strong intensities (> 1+) on both HIV-2 specific bands (sgp105 and gp36) on Innolia. CD4 counts were not significantly different in dual seropositives as compared to HIV-1 infected patients. CONCLUSIONS: Dual HIV-1/HIV-2 seropositives (and possibly infections) maybe common especially in older women. The Genie II will be useful as a supplemental rapid test for rapid and accurate differentiation of HIV-1 and HIV-2 antibodies at treatment centres. PMID- 21644412 TI - Aetiology of diarrhoea among persons with HIV and their family members in rural Uganda: a community-based study. AB - OBJECTIVES: To identify pathogens associated with diarrhoea in HIV-infected persons and their HIV-uninfected family members. DESIGN: Prospective cohort study. SETTING: Rural community in eastern Uganda. SUBJECTS: Eight hundred and seventy nine HIV-infected adults (74% females and median age 35 years (IQR, 29 41) and 2771 HIV-uninfected family members (51% females and median age 11 years (IQR 6-16) were included. MAIN OUTCOME MEASURES: Using microscopy and culture, stools were tested for parasites, bacteria and bacterial-antimicrobial susceptibility. Logistic regression models, adjusting for age, CD4 cells, season, household clustering and use of safe-water system were used for relationships between pathogens, diarrhoea and HIV. RESULTS: Persons with HIV had similar pathogens in diarrhoeal (69%) and nondiarrhoeal stools (57%). Most diarrhoea was not associated with identifiable aetiology; the population attributable risk of diarrhoea for known diarrhoea pathogens was 32%. Enteric bacteria (19%), enteropathogenic or enterotoxigenic E. coli (8%), Aeromonas species (7%), Strongyloides stercoralis (8%) and Cryptosporidium parvum (5%). HIV-infected, stools had more Cryptosporidium parvum than HIV-uninfected (OR 2.64, 95% CI 1.43 4.87). Most bacteria were resistant to commonly used antimicrobials irrespective of HIV status. CONCLUSIONS: Irrespective of HIV-status, aetiologies of majority of their diarrhoea in Uganda cannot be identified by microscopy and culture. Bacterial pathogens isolated have high resistance to common antimicrobials. Empiric treatment should be tailored to local bacterial-resistance patterns. PMID- 21644413 TI - Pregnancy rates among female participants in phase I and phase IIA AIDS vaccine clinical trials in Kenya. AB - BACKGROUND: Female participants in AIDS candidate vaccine clinical trials must agree to use effective contraception to be enrolled into the studies, and for a specified period after vaccination, since the candidate vaccines' effects on the embryo or foetus are unknown. OBJECTIVES: To review data on female participants' pregnancy rates from phase I and IIA AIDS vaccine clinical trials conducted at the Kenya AIDS Vaccine Initiative (KAVI) and to discuss the challenges of contraception among female participants. DESIGN: Descriptive observational retrospective study. SETTING: KAVI clinical trial site, Kenyatta National Hospital and University of Nairobi, Kenya. SUBJECTS: Thirty nine female participants were enrolled into these trials. They received family planning counselling and were offered a choice of different contraceptive methods, as per the protocols. All contraception methods chosen by the participants were offered at the study site at no cost to the participant. RESULTS: Four women conceived during the study period when pregnancies were to be avoided. All four had opted for sexual abstinence as a contraceptive method, but reported having been coerced by their partners to have unprotected sexual intercourse. CONCLUSION: Abstinence is clearly not a reliable contraceptive option for women in developing-country settings. Effective female-controlled contraceptives, administered at the clinical trial site, may empower female participants to better control their fertility, leading to more complete clinical trial data. PMID- 21644414 TI - Nephrotoxicity of amphotericin B in the treatment of cryptococcal meningitis in acquired immunodeficiency syndrome patients. AB - OBJECTIVES: To describe the incidence of renal dysfunction, hypokalaemia and hypomagnesaemia in AIDS patients with cryptococcal meningitis and on amphotericin B treatment. Secondary objective was to determine all-cause mortality in the same group. DESIGN: Prospective, observational study. SETTING: Kenyatta National Hospital (KNH), Nairobi, Kenya. SUBJECTS: Seventy consecutive patients with AIDS and cryptococcal meningitis on amphotericin B. RESULTS: About 58.6% of the patients had at least 100% rise in the creatinine level. Thirty eight point six per cent of patients experienced a rise in serum creatinine of at least 50%. Ninty three per cent of the patients developed hypokalaemia and 80% had hypomagnesaemia at trough magnesium level. Only 54.3% of patients completed the intended 14-day treatment. Thirty point five per cent of patients died within the two week follow-up period. CONCLUSION: The incidences of amphotericin B associated nephrotoxicity, hypokalemia and hypomagnesaemia were high in this studied population. PMID- 21644415 TI - Knowledge gaps, attitude and beliefs of the communities about sickle cell disease in Eastern and Western Uganda. AB - BACKGROUND: The management of sickle cell disease (SCD) has remained insurmountable in developing countries such as Uganda, because most communities are not aware of it. OBJECTIVE: To determine knowledge gaps, attitudes and beliefs of the communities about sickle cell disease in Eastern and Western Uganda. DESIGN: Cross sectional descriptive study. SETTING: The districts of Sironko and Mbale in Eastern Uganda and Mbarara and Ntungamo in Western Uganda. SUBJECTS: Households, students and health workers. RESULTS: Household respondents from Eastern Uganda were more aware of SCD than those from Western (p < 0.001), with the majority reporting that they had seen more people with SCD in their communities than those from the West (p < 0.001). Fewer (< 1.9%) believed SCD was due to witch craft. Eight per cent of household respondents in Eastern believed it was a curse from God compared to 2% in the West. Less than 18% of the household respondents knew they could have children with SCD and (< 52%) of health workers knew SCD screening methods. Fewer (< 14%) of the health workers had participated in screening. Less than 20% of the respondents knew their sickle cell status. CONCLUSION: Respondents from Eastern Uganda were more aware of SCD than those from Western. Minority of the respondents knew their SCD status and few health staff knew how to screen it. There is need to sensitise communities and policy makers about prevention, screening and treatment of SCD. PMID- 21644416 TI - Diagnostic values of digital rectal examination, prostate specific antigen and trans-rectal ultrasound in men with prostatism. AB - OBJECTIVE: To determine the diagnostic values of digital rectal examination (DRE), prostate specific antigen (PSA) and trans-rectal ultrasound (TRUS) individually and in combinations in men aged 50 years and above presenting with prostatism. DESIGN: A prospective, descriptive, cross-sectional, hospital-based study. SETTING: The urology ward of Kilimanjaro Christian Medical Centre (KCMC), a 500 bed tertiary hospital in the Kilimanjaro region, Tanzania. SUBJECTS: Ninety four consecutive admissions of men aged 50 years and above admitted with urinary symptoms suggestive of prostatism. MAIN OUTCOME MEASURES: Primary outcome measures included race and age of patient; Positive predictive values, sensitivities and specificities for DRE, PSA and TRUS individually and in combinations and histology of the prostate specimens submitted after Tru-cut, TURP or open prostatectomy. The secondary outcome measures were mean PSA and PSA density. RESULTS: There was a prostate adenocarcinoma incidence of 25.5%; all found among patients with PSA levels greater than 10 ng/ml. The positive predictive value (PPV), sensitivity and specificity of DRE for prostate cancer were 0.67%, 66.7% and 88.6% with an accuracy of 82.8%; while for TRUS, the respective values were 0.58%, 58.3% and 85.7% with an accuracy of 78.7%. PSA alone had a positive predictive value of 0.16. A combination of abnormal DRE and PSA (more than 4.0 ng/ml) had a positive predictive value (PPV) of 0.75 while when DRE, TRUS and PSA were all abnormal, the PPV rose to 0.80. CONCLUSION: A combination of DRE and PSA yields 75% diagnostic sensitivity for prostate cancer and is reliable enough to exempt TRUS where not available since it only adds 5% to this strong diagnostic combination. PMID- 21644417 TI - Congenital cystic adenomatoid malformation: case report. AB - Congenital cystic adenomatoid malformation of the lung is a rare lesion that typically manifests as severe progressive respiratory distress in the neonate secondary to expansion of the affected lung. We present a neonate in whom this condition was diagnosed and managed at the Aga Khan University Hospital. In presenting this patient, we hope to highlight the radiological aspects of this condition which, with prompt recognition and management and in the absence of pulmonary hypoplasia, foetal hydrops or other congenital anomalies, has a favourable prognosis. PMID- 21644418 TI - Induction of labour with titrated oral misoprostol suspension. A comparative study with vaginal misoprostol. AB - OBJECTIVE: To compare the effectiveness of titrated orally and vaginally administered misoprostol for induction of labour. STUDY DESIGN: Unmasked randomized controlled trial. SETTING: Department of Obstetrics and Gynaecology University of Zimbabwe, Harare. SUBJECTS: Pregnant women with singleton foetus in cephalic presentation booked for induction of labour, were randomized to receive titrated orally or vaginally administered misoprostol. MAIN OUTCOME MEASURES: The main outcomes were the duration of labour and induction to delivery interval. The secondary outcomes were neonatal and maternal complications. METHODS: 134 women were recruited into the study; 69 and 65 were randomized into orally and vaginally administered misoprostol respectively. RESULTS: The baseline characteristics in the two groups were similar. Women induced with titrated oral misoprostol suspension had a shorter interval from administration of the drug to initiation of uterine contractions (OR = 0.94 .95% CI 0.42 to 2.12) and a longer duration of labour (OR = 0.36; 95% CI 0.16 to 0.79). Labour was augmented with oxytocin in the oral group. The mean drug dose was 28mcg in the oral group. There was no difference in the mode of delivery between the two groups. Hypertonic uterine contractions were not detected. Ruptured uterus did not occur in the study population. There were more neonatal admissions in the vaginal than the oral group (OR = 1.03 .95% CI 0.29 to 1.39). CONCLUSION: Titrated oral misoprostol suspension is as effective and safe as vaginal misoprostol for induction of labour even in poor resource countries where intrapartum monitoring is inadequate. PMID- 21644419 TI - Mortality in an ophthalmic ward at a tertiary eye unit in Zimbabwe. AB - OBJECTIVE: To review and determine the attributed causes of deaths among patients who died in Sekuru Kaguvi Hospital (SKH) Eye Unit Wards between August 1984 and October 2008 STUDY DESIGN: Retrospective study of completed BD12 forms and case records of inpatients who died betweenAugust 1984 and October 2008. SETTING: Sekuru Kaguvi Hospital Eye Unit, Parirenyatwa Group ofHospitals, Harare, Zimbabwe. RESULTS: Eighty three deaths occurred in the unit during the 24 year period, 51% were females and 33.7% were children below 13 years of age. Ninety three percent of deaths among children were attributed to neoplasias, while 22% of deaths among adults were attributed to infections. Non-ophthalmic causes of deaths were seen among adults. CONCLUSION: Malignancies are the commonest attributable causes of mortality in the Ophthalmic Wards at Sekuru Kaguvi Hospital Eye Unit. Resource mobilization is essential to improve Eye Care Services. PMID- 21644420 TI - Knowledge, attitudes and practices on infant feeding options among HIV positive mothers. AB - OBJECTIVES: To determine knowledge, attitudes and practices of HIV positive mothers on infant feeding options. DESIGN: Descriptive cross-sectional survey. SETTING: A rural district hospital in Zimbabwe. SUBJECTS: A convenience sample of fifty HIV positive mothers attending Prevention of Mother-to-Child Transmission Programme with children under two years of age, were recruited for the study. MAIN OUTCOME MEASURES: Selected indicators of knowledge, attitudes and practices were analysed using descriptive statistics. RESULTS: The study revealed that the majority, 44 (88%), were able to define exclusive breastfeeding, though 36 (74%) practiced exclusive breastfeeding. On the dangers of mixed feeding, 36 (74%) were knowledgeable. Of those practicing exclusive breastfeeding, 25 (50%) were afraid of transmitting HIV virus to their babies, but were constrained by socio-economic and cultural factors. Forty five (90%) of the subjects suggested that husbands or partners should join counseling sessions on infant feeding options. CONCLUSION: Couple quality counseling is needed to assist in coping with challenges involved in infant feeding options. Breastfeeding support groups are needed after discharge. The size of the sample was small. However the, findings are important to clinical practice. PMID- 21644421 TI - Heterotopic pregnancy a case report. AB - A 27 year old patient was admitted with heterotopic pregnancy. Ultrasound scan done by her general practitioner had shown a 14 week viable intra-uterine pregnancy and fluid in the paracolic gutters. An exploratory laparatomy was done and she was found to have a ruptured right-sided ectopic pregnancy. Right partial salpingectomy was done. She had an uneventful post operative course. Ultrasound scan done at 18 weeks showed that the foetus was growing well. PMID- 21644422 TI - Treatment outcomes of patients on antiretrovirals after six months of treatment, Khami Clinic, Bulawayo, Zimbabwe. AB - OBJECTIVE: To describe treatment outcomes of patients on anti-retrovirals at six months of treatment. STUDY DESIGN: We conducted pre-intervention post intervention surveys using a pretest-post test design. SETTING: Khami Municipal Clinic, Bulawayo. SUBJECTS: We interviewed consecutive patients eligible to receive antiretroviral drugs (ARVs). All patients had a history of TB treatment and a CD4 count less than 200 cells/mm. MAIN OUTCOME MEASURES: Mean change in CD4 count, weight, body mass index, and Karnofsky performance measured before and at six months ofantiretroviral treatment. RESULTS: 72 subjects were interviewed at baseline, their median age was 38 years (Q1, 32 years, Q3, 43 years). Of these, 17 (24%) died before six months of treatment. Three (4%) defaulted treatment follow up. A total of 52 respondents were alive and interviewed at six months though only 50, had repeat CD4 counts at six months. Among the 50 survivors, the mean CD4 count at six months was significantly higher than at baseline (p = 0.0003). There was a 4.2 point statistical significant increase in the mean weight from baseline (p = 0.0005). Similarly, the mean Body Mass Index (BMI) significantly increased by 1.5 kg/m2 from baseline, (p = 0.001). The mean Karnofsky performance increased from 89% at baseline to 95% at six months (p = 0004). The researchers noted that patients on TB treatment were being deferred antiretroviral therapy until they completed TB treatment. CONCLUSION: The Khami project bears testimony that even in a resource poor setting; treatment of HIV/AIDS with antiretroviral drugs is feasible. We recommend early treatment initiation for those on TB treatment in line with national guidelines. PMID- 21644423 TI - Epidemiology of oral Kaposi's sarcoma in Zimbabwe 1988-1997: a population-based study. AB - OBJECTIVE: Sub-Saharan Africa has the highest number of HIV/AIDS cases globally which contrasts with the lack of population-based studies of oral Kaposi's sarcoma (OKS); one of the clinical cardinal signs of HIV/AIDS. To date, no study has investigated the incidence of OKS in African populations affected by the HIV/AIDS epidemic. It is, therefore, the purpose of this study to assess the burden of OKS in the Zimbabwean population over a 10 year period. DESIGN: A descriptive epidemiological study was undertaken to assess the burden of OKS by determining the frequencies, incidence and cumulative rates, the lifetime risk and chances of developing OKS according to site (topography), gender, age, race/ethnic origin of the Zimbabwean population. A total of 445 incident cases of OKS from the upper and lower lips, oral vestibule, retromolar area, floor of mouth, tongue, cheek, mucosa, gums, hard and soft palate were accessed from the Zimbabwe National Cancer Registry (ZNCR). Cases from the skin, pharynx, larynx and the major salivary glands were excluded from the study. SETTING: This comprised the population of Zimbabwe during the 10 year period 1988-1997. The population figures used for this study were fro the 1992 Census Zimbabwe National Report. The study population was standardized by the direct method against the world standard population to calculate the age standardized incidence rate (ASIR). The SPSS statistical software programme (SPSS Inc. 2001, USA) was used for the statistical analysis. RESULTS: OKS comprised 0.92% of total body malignancies and 51% of oral malignancies with a mean age of study cases of 37.6 years and median age of32 years. Histology of the primary (64.5%) and clinical diagnosis (34.6%) were the predominant methods of diagnosis. OKS affected nearly only blacks and males more than females, with a male to female ratio of 1.9:1. The most affected age groups by OKS were the 30 to 34 years for male and 25 to 29 years for both females and the whole population. Other notable peaks in OKS rates were in the 0 to 4 year and the 75+ age groups. OKS mostly affected the palate (70.2%) followed by, in descending order, the tongue (13.3%) and mouth (8.3%). The age adjusted age standardized incidence rate (ASIR) of OKS exponentially increased the entire study period bypassing oral squamous cell carcinoma (OSCC) as the predominant oral malignancy in 1994. Among AIDS-associated malignancies, OKS accounted for 98% while the balance comprised Burkitt's lymphoma, Hodgkin's and Non-Hodgkin's lymphomas, haemangiosarcoma and lymphoma not specified. CONCLUSION: OKS was the commonest malignancy of young adults affecting males more than females. OKS steady increased for the entire study period overtaking SCC in 1994 to become the commonest oral malignancy for the remainder of the study period. The palate was the most affected intra-oral site by OKS. These findings are attributable to the high human immunodeficiency virus infection (HIV) rates recorded for PMID- 21644424 TI - Malaria in pregnancy. PMID- 21644425 TI - At the crossroads: accountable care organizations and antitrust law. PMID- 21644427 TI - Selecting barrenness: the use of preimplantation genetic diagnosis by congenitally infertile women to select for infertility. AB - Congenitally infertile women such as those with Turner syndrome or Mayer Rokitansky-Kuster-Hauser syndrome have available the technologies of oocyte harvesting, cryropreservation, in-vitro fertilization, and gestational surrogacy in order to have genetically related offspring. Since congenital infertility results in a variety of experiences that impacts on nearly every aspect of a person's life, in the future it is possible that these women might desire a congenitally infertile child through the use of preimplantation genetic diagnosis so as to share this common bond. While infertility results in a relatively normal quality of life, it is morally wrong to necessitate the future use of infertility services with its variable success rate on a child. Also, whereas the woman has fundamental reproductive autonomy, she lacks the substantive autonomy regarding the specific characteristics of her child. Finally, the infertile community does exhibit a strong presence, but it lacks characteristics that define it as a culture. PMID- 21644426 TI - Retirement income adequacy with immediate and longevity annuities. PMID- 21644428 TI - Global principles, local obligations: reproductive ethics in affluent societies and developing countries. AB - This essay is an intercultural dialogue in reproductive ethics. The paper, which argues from both developed and developing world perspectives, addresses the question of what should be done when confronted with the possibility of giving birth to a severely disabled child. The author argues that such a life should not be considered because of the economic circumstances in most developing countries. This is contrary to the view sometimes advanced in affluent societies that the prevention of such a birth should not necessarily be considered. The author, however, agrees that the principle of acceptable outlook could be employed in both economic settings but with a variable degree of moral compliance without suggesting that certain lives are better than others. PMID- 21644429 TI - Stem cells, nuclear transfer and respect for embryos. AB - Harvesting human embryonic stem (hES) cells is a highly controversial field of research because it rests on the destruction of human embryos. Altering the procedure of nuclear transfer (NT) is suggested to generate hES cell lines without ethical obstacles by claiming that no embryo would be involved. While discussing the nature of an embryo and related central questions concerning their moral status and the respect they deserve, this paper argues that the entity created by somatic cell nuclear transfer (SCNT) or altered nuclear transfer (ANT) is an embryo and has the same moral status as a natural embryo. Respect for the embryo is expressed by the ethical principles of proportionality, probability and subsidiarity. This paper argues that the human embryo should only be taken for research with high ranking goals, which are proven in animal experimentation and for which there are no alternatives. This makes ANT obsolete and shows that SCNT to produce hES cells is premature at the present time. PMID- 21644430 TI - Therapeutic cloning: the ethical road to regulation. Part II: Analysing the UK position. PMID- 21644431 TI - On nature and bioethics. AB - The account of nature and humanity's relationship to nature are of central importance for bioethics. The Scientific Revolution was a critical development in the history of this question and many contemporary accounts of nature find their beginnings here. While the innovative approach to nature going out of the seventeenth century was reliant upon accounts of nature from the early modern period, the Middle Ages, late-antiquity and antiquity, it also parted ways with some of the understandings of nature from these epochs. Here I analyze this development and suggests that some of the insights from older understandings of nature may be helpful for bioethics today, even if there can be no simple return to them. PMID- 21644432 TI - Is the creation of admixed embryos "an offense against human dignity"? AB - The controversy over the creation of admixed human-nonhuman embryos, and specifically of what have been termed "cybrids," involves a range of ethical and political issues. It is not reducible to a single question. This paper focuses on one question raised by that controversy, whether creating admixed human-nonhuman entities is "an offense against human dignity. "In the last decade there has been sustained criticism of the use of the concept of human dignity within bioethics. The concept has been criticized as "vague" and "useless." Nevertheless, the concept continues to be invoked in bioethical discussion and in international instruments. This paper defends a concept of human dignity that is coherent but that is wider than contemporary post-Kantian approaches. "Human dignity" is best regarded as having a set of analogically related meanings, more than one of which is relevant to the field of bioethics. A more subtle understanding of the concept of human dignity can help identify what is ethically problematic in human nonhuman combinations and so shed light on one aspect of the admixed embryo debate. PMID- 21644433 TI - A matter of health? Legal aspects of private umbilical cord blood banking. AB - Private umbilical cord blood banking raises a question of special legal regulation. This practice promises the safe storage of biological material on the assumption that it may be useful, at a certain moment in future, for its own donor (or for a donor's close family member) for curing serious blood diseases. Although currently the therapeutic value of umbilical cord blood is confirmed, there are strong scientific doubts and relevant controversies regarding its use in autologous transplantations. This fact produces conditions of legal uncertainty, since the benefit for those wanting to conclude contracts with private umbilical cord blood banks is not clear. The Greek example illustrates this situation of regulatory deficit well, which eventually creates a major problem, given the increasing number of private banks offering relevant services in the country. PMID- 21644434 TI - Concurrence between complaints procedures in the Dutch healthcare system. AB - There are various ways in which complaints about healthcare providers' performance can be handled in The Netherlands. In practice, situations occur relatively frequently in which one and the same treatment by a provider results in various different complaints procedures being instigated or, in other words, in concurrence between complaints procedures. Dutch periodicals published information on a total of 42 cases of concurrence between 1997 and 2007. Concurrence between complaints procedures prompts a series of questions about how it arises, its legal implications, the possible consequences for those involved and whether it has any added value. This article examines these various issues. PMID- 21644435 TI - The way assisted suicide is legalised: balancing a medical framework against a demedicalised model. AB - To date, in three European countries and three American states--i.e., The Netherlands, Luxemburg, Switzerland, and the states of Oregon, Washington and Montana--it is permitted by law for one person to assist in the suicide of another person. When comparing the legislations of these countries/states, it becomes apparent that The Netherlands, Luxemburg, Oregon, Washington and Montana have chosen a medical approach (the so-called medical model), whereas the Swiss legal framework for assisted suicide is clearly a non-medical one (the demedicalised model). The differences between these two models mainly concern two aspects: the requirement as to the capacity of the person providing assistance in suicide and the condition regarding the state of health of the person committing suicide. A closer view on the practice of assisted suicide in the depenalising countries shows that the differences are smaller than initially thought. Nevertheless, important distinctions still remain. When analysing which model is most preferable, it is concluded that an involvement of a physician is inevitable and necessary and that the requirement of a certain medical condition is needed to set a clear and objective limit. PMID- 21644436 TI - Pushing the boundaries of lawful assisted dying in The Netherlands? Existential suffering and lay assistance. AB - Two matters that have a significant presence in the contemporary Dutch assisted dying debate, are the nature of the suffering required for an assisted death to be lawful, and the issue of who can lawfully assist. This article explores whether the lawful medical assisted dying model is too restrictive in failing to recognise existential suffering, considering selected case studies involving such suffering and lay assisted death. It addresses the question whether The Netherlands would take a trip down a slippery slope if the lawful model of assisted death were extended to cases where individuals are 'tired of life'. PMID- 21644437 TI - Introduction of a national electronic patient record in The Netherlands: some legal issues. AB - The electronic patient record (EPR) is a major technological development within the healthcare sector. Many hospitals across Europe already use institution-based electronic patient records, which allow not only for electronic exchange of patient data within the hospital, but potentially also for sharing medical data with external healthcare providers, involved in the patient's care, such as general practitioners or pharmacists. In this article, we discuss the attempt made by the Dutch government to introduce a nationwide electronic patient record (n-EPR). Describing and analyzing the new legislation that is currently being developed to establish the infrastructure for the n-EPR and the related legal issues, we conclude that the introduction of a n-EPR give rise to some substantial concerns. These vary from technical and quality issues such as the reliability of patient data and sufficient standardization and interoperability of the systems used, to issues in the field of data security and confidentiality. For a successful introduction of the n-EPR within the healthcare sector, a condicio sine qua non is that the related legislation provides sufficient safeguards and clarity with respect to the responsibilities and liabilities of its main users: the healthcare professionals. PMID- 21644438 TI - As easy as A, B and C: will A, B and C v. Ireland be Ireland's wake-up call for abortion rights? AB - This article examines the development of Ireland's abortion policy from 1861 to the present day. It explores the reasoning for this policy as well as the inherent problems with this policy. It examines in detail the A, B and C v. Ireland judgement and its impact, (if any) on Irish abortion law. Finally, it discusses the margin of appreciation doctrine used by the European Court of Human Rights in deciding cases of a moral nature. PMID- 21644439 TI - European Court of Human Rights. ECHR 2011/6 Case of Ternovszky v. Hungary, 14 December 2010, no. 67545/09 (Second Section). PMID- 21644440 TI - European Court of Human Rights. ECHR 2011/7 Case of A, B and C v. Ireland, 16 December 2010, no. 25579/05 (Grand Chamber). PMID- 21644441 TI - European Court of Human Rights. ECHR 2011/8 Case of Gladkiy v. Russia, 21 December 2010, no. 3242/03 (First Section). PMID- 21644442 TI - European Court of Human Rights. ECHR 2011/9 Case of Hoffer and Annen v. Germany, 13 January 2011, nos. 397/07 and 2322/07 (Fifth Section). PMID- 21644443 TI - European Court of Justice. ECJ 2011/4 18 November 2010 (C-156/09). PMID- 21644444 TI - European Court of Justice. ECJ 2011/5 9 December 2010 (C-421/09). PMID- 21644445 TI - A new era. "What can I know? What ought I to do? What may I hope?". PMID- 21644446 TI - Social-emotional functioning of elementary-age deaf children: a profile analysis. AB - Discussion and study of the social-emotional development of deaf and hard of hearing children, though extensive, has yet to provide an accurate understanding of the differences between deaf and hearing children. Consequently, the goal of the researchers was to conduct a profile analysis to determine similarities and differences between the two groups. The sample consisted of 20 hearing and 20 deaf children ages 8-11 years. All of the deaf children were enrolled in a Simultaneous Communication magnet program. Significant differences were found in two areas: school interest and on-task behavior. Overall, however, data from the study showed few differences between hearing and deaf children. The researchers recommend that current interventions be reconsidered on the basis of these results. PMID- 21644447 TI - Adapted interactive writing instruction with kindergarten children who are deaf or hard of hearing. AB - The study describes an adapted form of interactive writing (McCarrier, Pinnell, & Fountas, 2000) and examines its effectiveness as an approach to beginning writing instruction for young children who are deaf or hard of hearing. Systematic videotape analysis was used to document the content of 45 adapted interactive writing lessons across an academic year. Findings of the study suggest that interactive writing has the potential to support early writing development in young deaf and hard of hearing children, if supplemented by techniques that make the phonology of English visible. PMID- 21644448 TI - Faculty needs, doctoral preparation, and the future of teacher preparation programs in the education of deaf and hard of hearing students. AB - The purposes of the study were to obtain and analyze data on the need for, and desired characteristics of, faculty in deaf education at American institutions of higher education (IHEs), and to assess the present and projected status of doctoral-level teacher preparation programs in deaf education at American IHEs. Program directors and coordinators provided information on current and projected faculty openings, the number of active doctoral students, faculty research interests, program strengths, and needs in the field. Results indicated a pending shortage due to faculty retirements and a paucity of doctoral-level graduates. Most faculty listed literacy and language as a primary research interest as well as a program strength. The ability to generate new knowledge through research was found to be less desirable for future faculty than teaching ability. Suggestions for improving doctoral preparation and moving the field to evidence-based practices are provided. PMID- 21644449 TI - Directionality effects in simultaneous language interpreting: the case of sign language interpreters in The Netherlands. AB - The quality of interpretations produced by sign language interpreters was investigated. Twenty-five experienced interpreters were instructed to interpret narratives from (a) spoken Dutch to Sign Language of The Netherlands (SLN), (b) spoken Dutch to Sign Supported Dutch (SSD), and (c) SLN to spoken Dutch. The quality of the interpreted narratives was assessed by 5 certified sign language interpreters who did not participate in the study. Two measures were used to assess interpreting quality: the propositional accuracy of the interpreters' interpretations and a subjective quality measure. The results showed that the interpreted narratives in the SLN-to-Dutch interpreting direction were of lower quality (on both measures) than the interpreted narratives in the Dutch-to-SLN and Dutch-to-SSD directions. Furthermore, interpreters who had begun acquiring SLN when they entered the interpreter training program performed as well in all 3 interpreting directions as interpreters who had acquired SLN from birth. PMID- 21644450 TI - Integrating technology and reading instruction with children who are deaf or hard of hearing: the effectiveness of the Cornerstones project. AB - In a comparison between the Cornerstones approach--a literature-based, technology infused literacy project--and an instructional method designated the Typical approach, a mixed-method design was used to answer three research questions: (a) Will children who are deaf or hard of hearing demonstrate differences in beginning reading skills as measured by three outcome variables: Identification of Words in Print (or Word Identification), Word Knowledge, and Story Comprehension? (b) Are there carryover effects from the Cornerstones approach to the use of the Typical approach in subsequent experiments? (c) What is the feasibility of using the Cornerstones approach for literacy instruction? There were significant differences between the Typical and Cornerstones approaches in Word Identification and Story Comprehension in Experiments 1 and 2, though none in Word Knowledge or Story Comprehension in Experiment 3. Teacher feedback provided some evidence for the feasibility of using Cornerstones in the classroom. PMID- 21644451 TI - Enhancing our community of inquiry. Thoughts on principles and best practices in research with deaf and hard of hearing individuals. AB - The beginning of a new editorship for the Annals provides a good opportunity to re-engage contributors and the readership in a conversation about the principles and standards for manuscripts submitted to the Annals. In turn, the hope is that this conversation might help to enhance our research community and therefore the Annals' contributions to knowledge and understanding across an array of research and areas of inquiry focused on d/Deaf and hard of hearing individuals. PMID- 21644452 TI - A spatial dimension of ecology: Ilkka Hanski Crafoord laureate in 2011. PMID- 21644453 TI - Habitat loss, the dynamics of biodiversity, and a perspective on conservation. PMID- 21644454 TI - When ecosystem services crash: preparing for big, fast, patchy climate change. AB - Assessments of adaptation options generally focus on incremental, homogeneous ecosystem responses to climate even though climate change impacts can be big, fast, and patchy across a region. Regional drought-induced tree die-off in semiarid woodlands highlights how an ecosystem crash fundamentally alters most ecosystem services and poses management challenges. Building on previous research showing how choice of location is linked to adaptive capacity and vulnerability, we developed a framework showing how the options for retaining desired ecosystem services in the face of sudden crashes depend on how portable the service is and whether the stakeholder is flexible with regard to the location where they receive their services. Stakeholders using portable services, or stakeholders who can move to other locations to obtain services, may be more resilient to ecosystem crashes. Our framework suggests that entering into cooperative networks with regionally distributed stakeholders is key to building resilience to big, fast, patchy crashes. PMID- 21644455 TI - Climate warming and the recent treeline shift in the European alps: the role of geomorphological factors in high-altitude sites. AB - Global warming and the stronger regional temperature trends recently recorded over the European Alps have triggered several biological and physical dynamics in high-altitude environments. We defined the present treeline altitude in three valleys of a region in the western Italian Alps and reconstructed the past treeline position for the last three centuries in a nearly undisturbed site by means of a dendrochronological approach. We found that the treeline altitude in this region is mainly controlled by human impacts and geomorphological factors. The reconstruction of the altitudinal dynamics at the study site reveals that the treeline shifted upwards of 115 m over the period 1901-2000, reaching the altitude of 2505 m in 2000 and 2515 m in 2008. The recent treeline shift and the acceleration of tree colonization rates in the alpine belt can be mainly ascribed to the climatic input. However, we point out the increasing role of geomorphological factors in controlling the future treeline position and colonization patterns in high mountains. PMID- 21644456 TI - Effects of natural disasters on conservation policies: the case of the 2008 Wenchuan earthquake, China. AB - Conservation policies are increasing in response to human-induced ecosystem degradation, but little is known about their interplay with natural disasters. Through an analysis of satellite imagery and field data we evaluated the impacts of a devastating earthquake on forest recovery and avoided forest loss estimated to have been obtained by two of the largest conservation programs in the world. Results show that more than 10% of the forests in Wenchuan County, Sichuan province, China were immediately affected by the 2008 earthquake, offsetting some gains in forest cover observed since the enactment of the conservation programs. But without the enactment of these conservation programs, the combined effects of human disturbance and earthquake-induced landslides could have severely reduced the region's forest cover. The continuation--and enhancement--of incentives for participation in conservation programs will be important for reducing the environmental impacts of the combined effects of human disturbance and natural hazards not only in the study area but also in many disaster-prone regions around the world. PMID- 21644457 TI - Payments for environmental services in Latin America as a tool for restoration and rural development. AB - Payments for Environmental Services (PES) can encourage projects that enhance restoration, production, and rural development. When projects promote differentiated systems by paying farmers for the provision of services, the application of PES requires evaluation of the environmental services provided by each system. We present evaluations of carbon stocks and biodiversity in pure and mixed native tree plantations in Costa Rica. To illustrate how monetary values can be assigned, we discuss a project that awarded PES to silvopastoral systems in Costa Rica, Nicaragua, and Colombia based on carbon stocks and biodiversity. PES can promote positive environmental attitudes in farmers. Currently this project is being scaled up in Colombia based on their positive experiences with PES as a tool to promote adoption. Compared to PES systems that include only one environmental service, systems that incorporate bundling or layering of multiple services can make sustainable land uses more attractive to farmers and reduce perverse incentives. PMID- 21644458 TI - Implications of rural-urban migration for conservation of the Atlantic Forest and urban growth in Misiones, Argentina (1970-2030). AB - Global trends of increasing rural-urban migration and population urbanization could provide opportunities for nature conservation, particularly in regions where deforestation is driven by subsistence agriculture. We analyzed the role of rural population as a driver of deforestation and its contribution to urban population growth from 1970 to the present in the Atlantic Forest of Argentina, a global conservation priority. We created future land-use-cover scenarios based on human demographic parameters and the relationship between rural population and land-cover change between 1970 and 2006. In 2006, native forest covered 50% of the province, but by 2030 all scenarios predicted a decrease that ranged from 18 to 39% forest cover. Between 1970 and 2001, rural migrants represented 20% of urban population growth and are expected to represent less than 10% by 2030. This modeling approach shows how rural-urban migration and land-use planning can favor nature conservation with little impact on urban areas. PMID- 21644459 TI - The role of ethnobotanical skills and agricultural labor in forest clearance: evidence from the Bolivian Amazon. AB - Research on the benefits of local ecological knowledge for conservation lacks empirical data on the pathways through which local knowledge might affect natural resources management. We test whether ethnobotanical skills, a proxy for local ecological knowledge, are associated to the clearance of forest through their interaction with agricultural labor. We collected information from men in a society of gatherers-horticulturalist, the Tsimane' (Bolivia). Data included a baseline survey, a survey of ethnobotanical skills (n = 190 men), and two surveys on agricultural labor inputs (n = 466 plots). We find a direct effect of ethnobotanical skills in lowering the extent of forest cleared in fallow but not in old-growth forest. We also find that the interaction between ethnobotanical skills and labor invested in shifting cultivation has opposite effects depending on whether the clearing is done in old-growth or fallow forest. We explain the finding in the context of Tsimane' increasing integration to the market economy. PMID- 21644461 TI - Tourism under climate change: will slow travel supersede short breaks? PMID- 21644460 TI - Habitat as architecture: integrating conservation planning and human health. PMID- 21644462 TI - Consequent effects of parasitism on population dynamics, food webs, and human health under climate change. PMID- 21644463 TI - World Wide Food Webs: power to feed ecologists. PMID- 21644464 TI - Trends in body mass of ducks over time: the hypotheses in Guillemain et al. revisited. PMID- 21644465 TI - The different aspects of leprosy chemotherapy. PMID- 21644466 TI - Reflections on the new WHO leprosy indicator: the rate of new cases with grade 2 disabilities per 100,000 population per year. PMID- 21644467 TI - Trends in new leprosy case detection over 57 years (1952-2008) in Yuxi, Yunnan Province of Southwest China. AB - BACKGROUND: Leprosy control and management in China was maintained via a vertical system. Despite substantial achievements in leprosy control in the past decades, leprosy has not been completely eliminated in several endemic areas of Yunnan Province, China. OBJECTIVES: To report the epidemiological trends and management of leprosy in Yuxi, Yunnan Province from 1952 to 2008. DESIGN: Diagnosis, control, and treatment data for 2223 leprosy cases detected from 1952 to 2008 were analysed. RESULTS: Two large-scale house-to-house surveys were launched in 1957-1958 and 1964-1965, and a remarkable number of new cases were identified during these two surveys. The overall prevalence rate of leprosy in the Yuxi region presented a roughly unimodal distribution between 1952 and 2008, with a peak (9.27 per 10000 population) in 1965. This reflects a combination of case detection and duration of treatment. Overall, the age distribution of the patients changed dramatically over the years, and there were only two childhood cases between 1995 and 2008 (both occurring in 1998). Nearly half of the total cases (49.1%) were classified as multibacillary leprosy type. With the introduction and ubiquitous coverage of the WHO multi-drug therapy (MDT) in this area, leprosy elimination was achieved in 1992. In recent years, the majority of cases (> 80%) were detected by passive approaches, and there is an increasing tendency to find multibacillary leprosy patients. CONCLUSIONS: Our results provide an overall profile of our 57-year effort regarding the leprosy control in the Yuxi region. The trend in detection of new cases in recent years suggested that the transmission of leprosy has stopped in the area or, at least, dramatically declined. PMID- 21644468 TI - Adverse effects of multi-drug therapy in leprosy, a two years' experience (2006 2008) in tertiary health care centre in the tribal region of Chhattisgarh State (Bastar, Jagdalpur). AB - OBJECTIVE: To assess the adverse effects of multi drug therapy (MDT) in leprosy patients. METHODS: A prospective and descriptive study carried out in Department of Dermatology, Government Medical College, Jagdalpur. The adverse effects were recorded on the personal record of every individual patient, filled during the course of treatment. RESULTS: 176 patient's records were analysed, looking for adverse effects. Among the 176 patients, 79 had adverse effects due to one or more components of MDT, 73 had adverse effects due to dapsone, eight due to rifampicin and 16 due to clofazimine. Mean (+/- SD) duration for the development of adverse effects from the start of therapy was 1.99 (+/- 0.69) months for dapsone, 36 (+/- 0.68) months for rifampicin and 7.13 (+/- 0.79) months for clofazimine. There was a significant (P < 0.05) correlation between adverse effects and low Body Mass Index (BMI). The suspected drug was stopped and an alternative regime started in nine patients; six had dapsone stopped, two had rifampicin stopped and one had clofazimine stopped. CONCLUSION: Adverse effects attributed to MDT are comparable to previous studies and we found that ADR due to Dapsone was very high but most of the ADR were managed by supportive treatment without replacing the suspected drug. PMID- 21644469 TI - In vitro and skin lesion cytokine profile in Brazilian patients with borderline tuberculoid and borderline lepromatous leprosy. AB - OBJECTIVE: We investigated the in vitro and skin lesions production of cytokines in non-treated borderline tuberculoid (BT) and borderline lepromatous (BL) patients. PATIENTS AND METHODS: Seven untreated, non-reactional BT patients and 12 untreated, non-reactional BL patients were studied. Levels of the cytokines IFN-gamma, IL-10, TGF-beta1 and TNF-alpha were measured in supernantant of peripheral blood mononuclear cells (PBMC) cultures, stimulated with specific M. leprae antigen (sonicated and whole). The cytokines iNOS, IL-10 and TGF-beta1 were detected by immunohistochemistry in skin biopsies. RESULTS: BT patients produced higher levels of IFN-gamma than BL patients; iNOS expression in skin lesions was also higher in BT patients. TGF-beta1 was detected in more cells in BL patients; IL-10 expression was similar in both groups. There was a negative correlation between iNOS and TGF-beta1 expression in skin biopsies, positive correlation between TGF-beta1 in skin lesions and bacillary index, as well as positive correlation between iNOS detected in skin biopsies and PBMC IFN-gamma production. CONCLUSIONS: The BT patients had a mainly a Th1-profile of cytokines in their skin lesions and BL patients had a Th2 profile. PMID- 21644470 TI - Detection of mutations in folp1, rpoB and gyrA genes of M. leprae by PCR- direct sequencing--a rapid tool for screening drug resistance in leprosy. AB - Conventional Mouse foot-pad (MFP) assay for screening drug resistance in M. leprae is cumbersome and time-consuming (approximately 6 to 12 months). Molecular targets for different anti-leprosy drugs have been well defined. Molecular tools for rapid detection of drug resistance in M. leprae have been standardised. A study to compare molecular methods with MFP assay in determining the drug susceptibility of M. leprae was carried out. METHODS: Forty Bacteriological Index (BI) positive patients of leprosy with clinical features of relapse (25), new cases (11) and defaulters (4) were included in the study. A skin biopsy was done and the samples were processed using both MFP assay and Molecular method. PCR assays were carried out to amplify, 388 bp of folP1 gene for dapsone resistance, 305 bp of rpoB gene for rifampicin resistance and 342 bp of gyrA gene for ofloxacin resistance, followed by direct DNA sequencing. RESULTS: Significant growth in the MFP test was obtained in only 28 out of 40 biopsies processed (70%). Ten of these isolates were dapsone resistant; one isolate showed combined resistance against dapsone, rifampicin and clofazimine. Amplification for all three genes was successful in all the 40 (100%) samples. Among folP1 products sequenced, six isolates showed mutations at 53 (or) 55 amino acid positions. Those strains which showed high-level resistance with two log growth in MFP test, and/or showed growth in passage had mutations in folp1 gene. No mutation was detected in rpoB and gyrA products. Thus no molecular evidence of Rifampicin resistance was found in the DNA isolated from biopsies. CONCLUSION: Thus PCR direct sequencing--the rapid and high sensitive molecular technique can be applied for detection of resistance against dapsone, rifampicin and ofloxacin in M. leprae, to over come the limitations of the conventional MFP assay. PMID- 21644471 TI - A randomised controlled trial assessing the effect of adding clarithromycin to rifampicin, ofloxacin and minocycline in the treatment of single lesion paucibacillary leprosy in Agra District, India. AB - AIM: To assess if there is any additional short and long-term effect of adding clarithromycin to rifampicin, ofloxacin and minocycline (ROM), the combination here after called C-ROM, in treating single lesion PB leprosy detected in the field. METHODS: 300 patients, detected on active search in Agra district, who had single lesion leprosy but no nerve thickening, were randomly allocated (using random number table) to two treatment groups, 151 to ROM and 149 to C-ROM. All th patients were given single dose of ROM or C-ROM and followed up every 6 months for disease status, cure rate, reaction and relapse. Survival analysis was used to compare relapse rate. RESULTS: The cure rate at 2 years was 93.1% in ROM and 91.4% in C-ROM group. By this time three relapses had occurred in the ROM group while two patients were found to have relapsed in the C-ROM group. Thus, there was no statistical difference in relapse rates (2.1% vs. 1.41%, P = 0.287) in the two groups. Long term observations over 3-5 years revealed nine relapses (five in ROM, four in C-ROM) giving relapse rate of 1.05/100 Person years in ROM and 0.90/100 person years in C-ROM group--again no significant difference was observed (P = 0.87). CONCLUSION: The study shows that addition of clarithromycin to ROM does not significantly improve the efficacy as measured in terms of cure rates and relapse rates in single skin lesion leprosy patients. PMID- 21644472 TI - Social participation of people affected by leprosy after discontinuation of multidrug therapy. AB - OBJECTIVES: To evaluate social participation of individuals completing treatment for leprosy and to describe the relationship with physical sequelae and sociodemographic characteristics. DESIGN: A cross-sectional, descriptive study was performed involving all leprosy affected people who were treated between 1998 and 2006 in Sao Jose do Rio Preto, Brazil. General and clinical data were collected and the subjects were assessed using the degree of physical disability of the WHO (DPD-WHO), the Eye-hand-foot (EHF) score, and the participation scale (PS), which measures social participation of individuals with stigmatising problems. RESULTS: Of the 335 people treated in the period, 223 (66.6%) were interviewed. Of these, 51.6% were women, the mean age was 54 years (+/- 15.7 years), 66.4% had up to 6 years of formal education, 42.6% worked and 26.9% were retired. There was a predominance of dimorphous (borderline) leprosy (39.9%). Participants' physical and mental health was considered good in the month preceding the interview in 50.2% and 59.2%, respectively. Disabilities according to the DPD-WHO and EHF scores affected 32% of the participants and restrictions in social participation occurred in 35.4%. Multivariate analysis demonstrated a significant association between restrictions in social participation and family income of less than three minimum salaries (US$ 160.50), associated diseases, hospitalisation within the previous year and physical disability. CONCLUSION: The prevalence of restrictions in social participation is high, even after completing treatment for leprosy. Physical disability, associated diseases, recent hospitalisation and low family incomes influence the social participation of these individuals. PMID- 21644473 TI - Relapse study in smear positive multibacillary (MB) leprosy after 1 year WHO multi-drug therapy (MDT) in Cebu, Philippines. AB - OBJECTIVE: To determine the frequency, time interval to relapse and the possible risk factors for relapse in multibacillary (MB) leprosy after 1 year's treatment with the standard multi-drug therapy (MDT). MATERIALS AND METHODS: Smear positive MB patients treated with MDT for 1 year were enrolled in a prospective relapse study between 1999 and 2005 at the Leonard Wood Memorial Center for Leprosy Research (LWM). After treatment completion, at yearly intervals, patients underwent slit-skin smear examination and were clinically monitored for possible signs of relapse. RESULTS: 300 patients were recruited, and by 2009, follow-up totaled 1,913 patient years, with a mean of 6.4 years per patient. Only one case of relapse was detected, with an absolute relapse rate of 0.3% (0.52 per 1000 patient-years at risk (PYAR)); among a subset with pre-treatment bacterial indices (BI) of > or = 4 +, the rate was 0.6%. Relapse occurred 7 years after MDT. CONCLUSION: These data provide strong evidence of the long-term efficacy of the one year WHO-MDT for multibacillary (MB) leprosy patients, even in those with a high initial BI. PMID- 21644474 TI - Qualitative assessment of medication adherence at an urban leprosy outpatient clinic in Hyderabad, India. AB - OBJECTIVES: The objective of this study was to measure medication adherence amongst outpatients attending an urban leprosy clinic in Hyderabad, India. DESIGN: In this study of observational design, the urine spot test and Morisky Scale questionnaire were concurrently used as qualitative measures of medication adherence. RESULTS: Fifty two patients met the inclusion criteria for this study; 13 patients (25%) were non-adherent according to the Morisky scale questionnaire and 17 patients (33%) according to the urine spot test. 48% of patients were non adherent on the basis of the urine spot test and/or the Morisky scale questionnaire. CONCLUSION: The results suggest that poor medication adherence remains an ongoing issue in the management of outpatients with leprosy. PMID- 21644475 TI - Bilateral medial fingernail dystrophy as a presenting feature in a patient with leprosy. PMID- 21644476 TI - Reasons for interrupting multidrug therapy against leprosy: the patients' point of view. PMID- 21644477 TI - Report on the sixth meeting of the IDEAL (Initiative for Diagnostic and Epidemiological Assays for Leprosy) consortium held in Beijing, China on 23-25 August 2010. PMID- 21644478 TI - Leprosy statistics 2009: some thoughts. PMID- 21644479 TI - Information overload in our polarized society. PMID- 21644480 TI - Association between presence of visible in-house mold and health-related quality of life in adults residing in four U.S. states. AB - Despite the broad use of health-related quality of life (HRQOL) as one of the measurements to assess health status and effectiveness of health care and interventions, the impact of in-house mold exposure on HRQOL is unknown. The study described in this article examined the relationship between presence of visible in-house mold (PVIM) and HRQOL among adults. Data were analyzed from the 2005 and 2006 Behavioral Risk Factor Surveillance System (BRFSS) surveys that consisted of a random cross-sectional sample of 18,356 adults in four states. The authors examined the relationship between PVIM and three important indicators of the HRQOL by logistic regression analyses. Their results suggest that PVIM is independently associated with the indicators of HRQOL including mentally unhealthy, physically unhealthy, and total unhealthy days. Therefore, implementation of appropriate measures at the household level to eliminate or reduce in-house mold may improve individuals' HRQOL. PMID- 21644481 TI - Impact of an urban healthy homes intervention. AB - In the study described in this article, the authors evaluated the impact of an urban Healthy Homes intervention that included educational home visits. Their one group, pre-post design used a structured interview at baseline and postintervention. The sample (N = 84) was comprised of low-income children younger than 18 years of age with an asthma diagnosis or with an asthmatic mother. Children were primarily male (62%), < or =10 years old (74%), and lived in a single family home (69%) with moisture (61%). Interventions included a personalized action plan, education, demonstrations, and home remediation as needed. Significant post-intervention decreases occurred in reported asthma symptoms for the child, school days missed, caregiver work days missed, and emergency department/urgent care center visits. Caregiver self-efficacy and quality of life increased significantly. The comprehensive home visitation intervention program effectively impacted asthma symptoms, lost school and work days, emergency use of the health care system, and improved caregiver quality of life and self-efficacy. PMID- 21644482 TI - Sources, distribution, and toxicity of polycyclic aromatic hydrocarbons. AB - Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous pollutants released from the incomplete combustion of fossil fuels and are always found as a mixture of individual compounds. Due to economic growth and a sharp increase in energy consumption in recent years, large quantities of PAHs have been released into the environment worldwide. Because many PAHs and their derivatives are strongly potent carcinogens, or mutagens, PAHs have been extensively studied recently. The authors reviewed the origin and distribution of PAHs in atmosphere, soil, and sediment in natural environments. PAHs represent a class of toxicological compounds that can create a variety of hazardous effects in vivo/in vitro, including genotoxicity, immunotoxicity, developmental toxicity, and carcinogenesis, which the authors also describe. PMID- 21644483 TI - The three Es of healthy swimming. PMID- 21644484 TI - The story. PMID- 21644485 TI - Surgical margins for melanoma: simple excision? PMID- 21644486 TI - What's eating you? Hyalomma ticks. PMID- 21644487 TI - What is your diagnosis? Molluscum contagiosum on the sole of the foot. PMID- 21644488 TI - Hodgkin lymphoma presenting as generalized pruritus in an adolescent. AB - Pruritus is a common manifestation of Hodgkin lymphoma (HL), and given its high frequency, inclusion of itching as a B symptom of HL has been proposed. We present a 16-year-old adolescent boy with treatment-refractory eczema of 2 years' duration. Physical examination revealed a thin adolescent boy with widespread excoriations, but no eczematous or primary cutaneous lesions were identifiable. Lymph node examination revealed palpably enlarged nodes in the cervical and supraclavicular regions. Laboratory studies revealed leukocytosis and an elevated lactate dehydrogenase level. Diffuse lymphadenopathy was detected on a chest radiograph, and excisional lymph node biopsy revealed HL (nodular sclerosing subtype). The patient was classified as HL stage IIIB (Ann Arbor staging classification) after further evaluation. Chemotherapy was initiated followed by radiation therapy. The patient's pruritus markedly improved within 2 cycles of chemotherapy; however, his HL relapsed and additional salvage combination chemotherapy followed by high-dose chemotherapy and autologous stem cell transplant were required. This case underscores the need for a complete history as well as a careful skin and systemic evaluation in patients presenting with long-term pruritus, including children and adolescents. PMID- 21644489 TI - Laptop computer-induced erythema ab igne on the left breast. PMID- 21644490 TI - Unusual presentation of a squamous cell carcinoma. PMID- 21644491 TI - Clopidogrel-associated acute generalized exanthematous pustulosis. AB - Acute generalized exanthematous pustulosis (AGEP) is a skin eruption of rapid onset and progression that is characterized by the formation of numerous sterile pustules on an erythematous background. Other features may include fever and leukocytosis, with resolution usually in less than 15 days. We present an 83-year old woman with an erythematous pustular eruption following placement of a bare metal coronary stent and initiation of anticoagulation with clopidogrel. She was diagnosed with AGEP after a biopsy was performed and other possible causes of a pustular eruption were excluded. Two medications--carvedilol and rosuvastatin- were initially viewed as possible culprits and were temporarily discontinued. However, the patient's rash continued to worsen. Additionally, she had a persistent low-grade fever, leukocytosis, and a slow rise in her creatinine level. Clopidogrel was discontinued and replaced with ticlopidine on day 7 of hospitalization. Within 48 hours her temperature, white blood cell count, and creatinine level began to improve. Her skin subsequently desquamated and she was sent home on day 20. We discuss drug-induced AGEP and a desensitization protocol that has allowed patients to resume use of clopidogrel following a prior cutaneous reaction to this medication. PMID- 21644492 TI - Scalp metastasis of paraspinal alveolar rhabdomyosarcoma. AB - We describe a 10-year-old girl with paraspinal alveolar rhabdomyosarcoma (RMS) who presented with an asymptomatic soft tissue mass on the left aspect of the parietal scalp of 2 weeks' duration. Biopsy demonstrated metastasis of her RMS. A full-body positron emission tomography (PET) scan revealed multiple areas of increased uptake consistent with extensive metastases. We also discuss the literature on RMS classification and metastasis to the skin. PMID- 21644493 TI - Stress and serum TNF-alpha levels may predict disease outcome in patients with pemphigus: a preliminary study. AB - The aim of the current preliminary case-control study was to estimate the initial serum levels of tumor necrosis factor alpha (TNF-alpha) in case patients with pemphigus vulgaris (PV) and pemphigus foliaceus (PF) and correlate them with history of stress, body surface area (BSA) affected, disease severity, and disease outcome. Ten PV and 4 PF case patients as well as 7 healthy matched controls had their serum levels of TNF-alpha measured by an enzyme-linked immunosorbent assay. Case patients were treated and followed up for 2 months. A statistically significant elevation in serum levels of TNF-alpha in PV case patients compared with controls and in PV case patients compared with PF case patients was detected (P < .05), with no significant difference between PF case patients and controls (P > .05). No significant correlation was detected between the serum levels of TNF-alpha and the BSA affected (P > .05). Four PV case patients had a bad disease outcome, of which 3 had severe emotional stress a month prior to the onset of the attack. All 4 showed significantly elevated initial serum levels of TNF-alpha compared with those who had a good disease outcome (P < .05). Emotional stress is a factor affecting prognosis of the disease. Pretreatment assessment of serum TNF-alpha levels in patients with pemphigus may be a guide to the expected prognosis and selection of the proper treatment regimen. PMID- 21644494 TI - Milia: a unique reaction to tattoos. AB - Tattoo-related dermatoses are varied and uncommon. Although rare, reactions to tattoos have been reported with a multitude of tattoo pigments and most commonly present with an eczematous reaction pattern. Milia are small keratinous cysts that may arise as primary lesions or secondary to some other trauma to the skin. We report the case of a 28-year-old man who presented with a papular eruption of 3 months' duration confined to the area of recently placed tattoos; the eruption was diagnosed as milia. PMID- 21644495 TI - Nonhealing scalp wound infected with Aspergillus niger in an elderly patient. AB - Cutaneous aspergillosis is a rare infection most often seen in immunocompromised patients. We report a case of primary cutaneous aspergillosis infection in a nonhealing scalp wound of an immunocompetent elderly patient. The patient had a cutaneous malignancy of the scalp treated with surgical excision but complicated by poor wound healing. Fungal culture of the nonhealing wound revealed Aspergillus niger. The nonhealing wound subsequently resolved with retapamulin ointment 1% and ketoconazole gel 2%. PMID- 21644496 TI - Progression of actinic keratosis to squamous cell carcinoma revisited: clinical and treatment implications. AB - Changes in the appearance of actinic keratosis (AK) suggest progression to invasive squamous cell carcinoma (SCC), though some dermatologists and dermatopathologists consider AK to be SCC in situ. Actinic keratosis is an indicator of cumulative UV exposure and the initial lesion in the majority of invasive cutaneous SCCs. The development of SCC on sun-damaged skin is a gradual process; however, most AK lesions do not progress to invasive SCC and it currently is not possible to clinically or histopathologically determine which AK lesions will progress to SCC. Presently there is insufficient evidence to support the concept that AK is frank SCC. Although the rate of progression over time remains to be determined by large prospective studies, AK is a marker for an increased rate of nonmelanoma skin cancer (NMSC), even in the absence of specific lesion progression. Nevertheless, the risk for progression of AK to invasive SCC with the potential for metastasis provides the rationale for treatment, and AK lesions should be treated with lesion- or field-directed therapy or with a combined approach when indicated. We discuss the implications for treatment and review a variety of treatment options. PMID- 21644497 TI - Toward a consensus view on the infectious risks associated with land application of sewage sludge. AB - The science linking processed sewage sludge (biosolids) land application with human health has improved in the last ten years. The goal of this review is to develop a consensus view on the human health impacts associated with land applying biosolids. Pre-existing risk studies are integrated with recent advances in biosolids pathogen exposure science and risk analysis. Other than accidental direct ingestion, the highest public risks of infection from land application are associated with airborne exposure. Multiple, independent risk assessments for enteroviruses similarly estimate the yearly probabilities of infection near 10( 4). However, the inclusion of other emerging pathogens, specifically norovirus, increases this yearly infectious risk by over 2 orders of magnitude. Quantitative microbial risk assessment for biosolids exposure more effectively operates as a tool for analyzing how exposure can be reduced rather than being used to assess "safety". Such analysis demonstrates that the tradition of monitoring pathogen quality by Salmonella spp. and enterovirus content underestimates the infectious risk to the public, and that a rigorous biosolids pathogen treatment process, rather than extending community separation distances, is the most efficient method for reducing pathogen exposure and infectious risk. PMID- 21644498 TI - High-pressure synthesis and correlation between structure, magnetic, and dielectric properties in LiNbO(3)-type MnMO3 (M=Ti, Sn). AB - LiNbO(3)-type MnMO(3) (M = Ti, Sn) were synthesized under high pressure and temperature; their structures and magnetic, dielectric, and thermal properties were investigated; and their relationships were discussed. Optical second harmonic generation and synchrotron powder X-ray diffraction measurements revealed that both of the compounds possess a polar LiNbO(3)-type structure at room temperature. Weak ferromagnetism due to canted antiferromagnetic interaction was observed at 25 and 50 K for MnTiO(3) and MnSnO(3), respectively. Anomalies in the dielectric permittivity were observed at the weak ferromagnetic transition temperature for both the compounds, indicating the correlation between magnetic and dielectric properties. These results indicate that LiNbO(3)-type compounds with magnetic cations are new candidates for multiferroic materials. PMID- 21644499 TI - A powerful new construction of complex chiral polycycles by an indium(III) catalyzed cationic cascade. AB - InI(3) and InBr(3) have been found to be effective catalysts for the pi activation of C=C bonds to initiate the conversion of chiral propargylic alcohols or silyl ethers to polycyclic products in excellent yields and with high stereoselectivity. The method has been applied to the synthesis of chiral fused hexacyclic ring systems with the creation of multiple new stereocenters. The power and scope of the method are illustrated by a variety of examples. PMID- 21644500 TI - Reactive facets covered mosaic spheres of anatase TiO2 and related pseudo isotropic effect. AB - Pseudo-isotropic mosaic spheres of anatase almost completely covered with reactive (001) facets are prepared via a one-pot reaction. The growth process is mediated by an appropriate amount of NaF under hydrothermal conditions. Due to the large exposure ratio of identical (001) facets, the product can serve as an ideal model to investigate certain crystalline plane effects owing to their pseudo-isotropic properties, such as texture effects. Intensified X-ray diffraction results are presented to show such effects induced by morphology. PMID- 21644501 TI - Enhancing the accuracy of chemogenomic models with a three-dimensional binding site kernel. AB - Computational chemogenomic (or proteochemometric) methods predict target-ligand interactions by training machine learning algorithms on known experimental data in order to distinguish attributes of true from false target-ligand pairs. Many ligand and target descriptors can be used for training and predicting binary associations or even binding affinities. Several chemogenomic studies have not noticed any real benefit in using 3-D structural target descriptors with respect to simpler sequence-based or property-based information. To assess whether this observation results from inaccurate target description or from the fact that 3-D information is simply not required in chemogenomic modeling, we used a target kernel measuring the distance between target-ligand binding sites of known X-ray structures. When used in combination with a standard ligand kernel in a support vector machine (SVM) classifier, the 3-D target kernel significantly outperforms a sequence-based target kernel in discriminating 2882 target-ligand PDB complexes from 9128 false pairs, whatever the modeling procedure (local or global). The best SVM models could be successfully applied to predict, with very high recall (70%), precision (99%), and specificity (99%), target-ligand associations for an external set of 14,117 ligands and 531 targets. In most of the cases, pooling all data in a global model gave better statistics than just discretizing specific target-ligand subspaces in local models. The current study clearly demonstrates that chemogenomic models taking both ligand and target information outperform simpler ligand-based models. It also permits one to design good modeling practices in predicting target-ligand pairing for a large array of targets: (i) ligand-based models are precise enough if sufficient ligand information (>40-50 diverse ligands) is known; (ii) if not, structure-based chemogenomic models (associating a ligand kernel to a structure-based target kernel) are recommended for proteins of known holostructures; (iii) sequence-based chemogenomic models (associating a ligand kernel to a sequence-based target kernel) can still be used with a very good accuracy for the remaining targets. PMID- 21644502 TI - Advances in the replacement and enhanced replacement method in QSAR and QSPR theories. AB - The selection of an optimal set of molecular descriptors from a much greater pool of such regression variables is a crucial step in the development of QSAR and QSPR models. The aim of this work is to further improve this important selection process. For this reason three different alternatives for the initial steps of our recently developed enhanced replacement method (ERM) and replacement method (RM) are proposed. These approaches had previously proven to yield near optimal results with a much smaller number of linear regressions than the full search. The algorithms were tested on four different experimental data sets, formed by collections of 116, 200, 78, and 100 experimental records from different compounds and 1268, 1338, 1187, and 1306 molecular descriptors, respectively. The comparisons showed that one of the new alternatives further improves the ERM, which has shown to be superior to genetic algorithms for the selection of an optimal set of molecular descriptors from a much greater pool. The new proposed alternative also improves the simpler and the lower computational demand algorithm RM. PMID- 21644503 TI - Extraction of discontinuous structure-activity relationships from compound data sets through particle swarm optimization. AB - The characterization of structure-activity relationship (SAR) features of large compound data sets has been a hot topic in recent years, and different methods for large-scale SAR analysis have been introduced. The exploration of local SAR components and prioritization of compound subsets have thus far mostly relied on graphical analysis methods that capture similarity and potency relationships in a systematic manner. A currently unsolved problem in large-scale SAR analysis is how to automatically select those compound subsets from large data sets that carry most SAR information. For this purpose, we introduce a numerical optimization scheme that is based on particle swarm optimization guided by an SAR scoring function. The methodology is applied to four large compound sets. We demonstrate that compound subsets representing the most discontinuous local SARs are consistently selected through particle swarm optimization. PMID- 21644504 TI - Optimized unlike-pair interactions for water-carbon dioxide mixtures described by the SPC/E and EPM2 models. AB - The unlike-pair interaction parameters for the SPC/E-EPM2 models have been optimized to reproduce the mutual solubility of water and carbon dioxide at the conditions of liquid-supercritical fluid phase equilibria. An efficient global optimization of the parameters is achieved through an implementation of the coupling parameter approach, adapted to phase equilibria calculations in the Gibbs ensemble, that explicitly corrects for the overpolarization of the SPC/E water molecule in the nonpolar CO(2) environments. The resulting H(2)O-CO(2) force field accurately reproduces the available experimental solubilities at the two fluid phases in equilibria as well as the corresponding species tracer diffusion coefficients. PMID- 21644505 TI - Comment on "Molecular selectivity in aquaporin channels studied by the 3D-RISM theory". PMID- 21644507 TI - Probabilistic consensus scoring improves tandem mass spectrometry peptide identification. AB - Database search is a standard technique for identifying peptides from their tandem mass spectra. To increase the number of correctly identified peptides, we suggest a probabilistic framework that allows the combination of scores from different search engines into a joint consensus score. Central to the approach is a novel method to estimate scores for peptides not found by an individual search engine. This approach allows the estimation of p-values for each candidate peptide and their combination across all search engines. The consensus approach works better than any single search engine across all different instrument types considered in this study. Improvements vary strongly from platform to platform and from search engine to search engine. Compared to the industry standard MASCOT, our approach can identify up to 60% more peptides. The software for consensus predictions is implemented in C++ as part of OpenMS, a software framework for mass spectrometry. The source code is available in the current development version of OpenMS and can easily be used as a command line application or via a graphical pipeline designer TOPPAS. PMID- 21644508 TI - beta-arrestin-2 regulation of the cAMP response element binding protein. AB - Previous work demonstrated that cystic fibrosis (CF) cells exhibit an increase in cAMP-mediated signaling as a characteristic response to lost CFTR function. Evidence for increased cAMP-mediated signaling in CF included increased phosphorylation of the cAMP response element binding protein (CREB) and elevated beta-arrestin-2 (betaarr2) expression. However, subsequent studies reveal that CREB activation in CF cells is independent of protein kinase-A (PKA). The goal of this study is to test the hypothesis that elevated betaarr2 expression leads to increased CREB activation in a PKA-independent mechanism. betaarr2-GFP expressing tracheal epithelial cells (betaarr2-GFP) exhibit an increase of pCREB content and subsequent CRE activation compared to GFP expressing control cells. betaarr2 activation of the ERK cascade represents a candidate mechanism leading to CREB activation. ERK exhibits increased activation in betaarr2-GFP cells compared to cont-GFP cells, and ERK inhibition diminishes CRE activation in both GFP and betaarr2-GFP cells. To test directly whether CREB regulation in CF is betaarr2 dependent, nasal epithelium excised from wt mice (Cftr +/+; betaarr2 +/+), CF mice (Cftr -/-; betaarr2 +/+), and DKO mice (Cftr -/-; betaarr2 -/-) were analyzed for pCREB protein content. Removal of betaarr2 expression from CF mice reduces both pCREB and pERK content to wt levels. These data indicate that CF related CREB regulation is mediated directly through betaarr2 expression via the ERK pathway. PMID- 21644509 TI - Combined serum and tissue proteomic study applied to a c-Myc transgenic mouse model of hepatocellular carcinoma identified novel disease regulated proteins suitable for diagnosis and therapeutic intervention strategies. AB - Hepatocellular carcinoma (HCC) is the third leading cause of cancer death in the U.S. Notably, most HCCs display c-Myc hyperactivity but this transcription factor participates in the regulation of as many as 15-20% of genes of the human genome. To better understand its oncogenic activity, a mass spectrometry-based proteomic approach was employed to search for disease-regulated proteins in liver tissue and serum of c-Myc transgenic mice that specifically developed HCC. Overall, a total of 90 differentially expressed proteins were identified with retinol binding protein 4, transthyretin, major urinary protein family, apolipoprotein E, and glutathione peroxidase being regulated in common in tissue and serum of HCC mice. Importantly, this study identified n = 22 novel tumor tissue-regulated proteins to function in cell cycle and proliferation, nucleotide and ribosomal biogenesis, oxidative stress, and GSH metabolism, while bioinformatics revealed the coding sequences of regulated proteins to enharbour c-Myc binding sites. Translation of the findings to human disease was achieved by Western immunoblotting of serum proteins and by immunohistochemistry of human HCC. Taken collectively, our study helps to define a c-Myc proteome suitable for diagnostic and possible therapeutic intervention strategies. PMID- 21644510 TI - Anaerobic biodegradation of longer-chain n-alkanes coupled to methane production in oil sands tailings. AB - Extraction of bitumen from mined oil sands ores produces enormous volumes of tailings that are stored in settling basins (current inventory >= 840 million m(3)). Our previous studies revealed that certain hydrocarbons (short-chain n alkanes [C(6)-C(10)] and monoaromatics [toluene, o-xylene, m-xylene]) in residual naphtha entrained in the tailings are biodegraded to CH(4) by a consortium of microorganisms. Here we show that higher molecular weight n-alkanes (C(14), C(16), and C(18)) are also degraded under methanogenic conditions in oil sands tailings, albeit after a lengthy lag (~180 d) before the onset of methanogenesis. Gas chromatographic analyses showed that the longer-chain n-alkanes each added at ~400 mg L(-1) were completely degraded by the resident microorganisms within ~440 d at ~20 degrees C. 16S rRNA gene sequence analysis of clone libraries implied that the predominant pathway of longer-chain n-alkane metabolism in tailings is through syntrophic oxidation of n-alkanes coupled with CO(2) reduction to CH(4). These studies demonstrating methanogenic biodegradation of longer-chain n-alkanes by microbes native to oil sands tailings may be important for effective management of tailings and greenhouse gas emissions from tailings ponds. PMID- 21644511 TI - Gold nanoparticles with cyclic phenylazomethines: one-pot synthesis and metal ion sensing. AB - New gold nanoparticles covered with cyclic phenylazomethine (CPA) were obtained by a one-pot synthesis. It is confirmed by XPS that imines of CPA in the nanoparticles (Au-CPA) are partially reduced to amines. The amine part of CPA in Au-CPA is attached to the surfaces of gold nanoparticles, and the imine part works as a redox-active site. A glassy carbon electrode modified with Au-CPA was revealed to work as an electrochemical probe for metal ion sensing. PMID- 21644512 TI - Switch in stereoselectivity caused by the isocyanide structure in the rhodium catalyzed silylimination of alkynes. AB - The reaction of terminal alkynes with hydrosilanes and tert-alkyl isocyanides in the presence of Rh(4)(CO)(12) gives (Z)-beta-silyl-alpha,beta-unsaturated imines in good yields. On the other hand, the use of aryl isocyanides in place of tert alkyl isocyanides leads to the formation of E isomers. PMID- 21644513 TI - Asymmetric synthesis of the core structure of leucosceptroids A-D. AB - The asymmetric synthesis of the core structure of leucosceptroids A-D has been achieved. The key steps of the synthesis includes the formation of the cis-2,5 disubstituted THF ring by TPAP catalytic oxidative cyclization followed by a highly diastereoselective intramolecular Diels-Alder reaction to fashion the fused tricyclic hydrindane ring system. PMID- 21644514 TI - Hamaker constants of iron oxide nanoparticles. AB - The Hamaker constants for iron oxide nanoparticles in various media have been calculated using Lifshitz theory. Expressions for the dielectric responses of three iron oxide phases (magnetite, maghemite, and hematite) were derived from recently published optical data. The nonretarded Hamaker constants for the iron oxide nanoparticles interacting across water, A(1w1) = 33 - 39 zJ, correlate relatively well with previous reports, whereas the calculated values in nonpolar solvents (hexane and toluene), A(131) = 9 - 29 zJ, are much lower than the previous estimates, particularly for magnetite. The magnitude of van der Waals interactions varies significantly between the studied phases (magnetite < maghemite < hematite), which highlights the importance of a thorough characterization of the particles. The contribution of magnetic dispersion interactions for particle sizes in the superparamagnetic regime was found to be negligible. Previous conjectures related to colloidal stability and self-assembly have been revisited on the basis of the new Lifshitz values of the Hamaker constants. PMID- 21644515 TI - Enhanced electrocatalytic performance of processed, ultrathin, supported Pd-Pt core-shell nanowire catalysts for the oxygen reduction reaction. AB - We report on the synthesis, characterization, and electrochemical performance of novel, ultrathin Pt monolayer shell-Pd nanowire core catalysts. Initially, ultrathin Pd nanowires with diameters of 2.0 +/- 0.5 nm were generated, and a method has been developed to achieve highly uniform distributions of these catalysts onto the Vulcan XC-72 carbon support. As-prepared wires are activated by the use of two distinctive treatment protocols followed by selective CO adsorption in order to selectively remove undesirable organic residues. Subsequently, the desired nanowire core-Pt monolayer shell motif was reliably achieved by Cu underpotential deposition followed by galvanic displacement of the Cu adatoms. The surface area and mass activity of the acid and ozone-treated nanowires were assessed, and the ozone-treated nanowires were found to maintain outstanding area and mass specific activities of 0.77 mA/cm(2) and 1.83 A/mg(Pt), respectively, which were significantly enhanced as compared with conventional commercial Pt nanoparticles, core-shell nanoparticles, and acid-treated nanowires. The ozone-treated nanowires also maintained excellent electrochemical durability under accelerated half-cell testing, and it was found that the area specific activity increased by ~1.5 fold after a simulated catalyst lifetime. PMID- 21644516 TI - Nanomaterials for X-ray imaging: gold nanoparticle enhancement of X-ray scatter imaging of hepatocellular carcinoma. AB - We present the development of a new imaging technique for the early diagnosis of hepatocellular carcinoma that utilizes surface-modified gold nanoparticles in combination with X-ray imaging. Tissues labeled with these electron-dense particles show enhanced X-ray scattering over normal tissues, distinguishing cells containing gold nanoparticles from cells without gold in X-ray scatter images. Our results suggest that this novel approach could enable the in vivo detection of tumors as small as a few millimeters in size. PMID- 21644517 TI - Dithienogermole as a fused electron donor in bulk heterojunction solar cells. AB - We report the synthesis and bulk heterojunction photovoltaic performance of the first dithienogermole (DTG)-containing conjugated polymer. Stille polycondensation of a distannyl-DTG derivative with 1,3-dibromo-N-octyl thienopyrrolodione (TPD) results in an alternating copolymer which displays light absorption extending to 735 nm, and a higher HOMO level than the analogous copolymer containing the commonly utilized dithienosilole (DTS) heterocycle. When polyDTG-TPD:PC(70)BM blends are utilized in inverted bulk heterojunction solar cells, the cells display average power conversion efficiencies of 7.3%, compared to 6.6% for the DTS-containing cells prepared in parallel under identical conditions. The performance enhancement is a result of a higher short-circuit current and fill factor in the DTG-containing cells, which comes at the cost of a slightly lower open circuit voltage than for the DTS-based cells. PMID- 21644518 TI - Layer-by-layer assembly of polysaccharide-based polyelectrolyte multilayers: a spectroscopic study of hydrophilicity, composition, and ion pairing. AB - Polyelectrolyte multilayers using the polycations chitosan and N,N,N-trimethyl chitosan and the polyanions hyaluronan, chondroitin sulfate, and heparin are studied. Chitosan and hyaluronan behave as a weak polycation and weak polyanion, respectively, whereas N,N,N-trimethyl chitosan, chondroitin sulfate, and heparin behave as strong polyelectrolytes. Hydrophilicity is determined by water contact angle measurements and by comparing wet and dry film thickness measurements. Wet thickness is obtained using Fourier transform surface plasmon resonance, whereas dry thickness is obtained through ellipsometry. For the very thin PEMs studied here, the surface hydrophilicity and swelling in water are highly correlated. The multilayer chemistry is assessed by FT-IR and X-ray photoelectron spectroscopy (XPS). FT-IR and XPS provide information about the composition, degree of ionization, and by inference, the ion pairing. We find that hydrophilicity and swelling are reduced when one polyelectrolyte is strong and the other is weak, whereas ion pairing is increased. By this combination of techniques, we are able to compose a unified description of how the PEM swelling is dictated by the ion pairing in thin polysaccharide-based PEMs. PMID- 21644519 TI - A chemoenzymatic total synthesis of (+)-clividine. AB - The title compound, ent-1, the non-natural enantiomeric form of the lycorenine type alkaloid (-)-clividine (1), has been prepared using the enantiomerically pure (ee >99.8%) cis-1,2-dihydrocatechol 3 as starting material. A key feature associated with the closing stages of the synthesis involved the diastereoselective addition of a nitrogen-centered radical onto a pendant cyclohexene to establish the cis-fused D-ring and the required stereochemistry at C11b in the final product ent-1. PMID- 21644520 TI - Hierarchically imprinted porous films for rapid and selective detection of explosives. AB - On the basis of the combination of colloidal and mesophase templating, as well as molecular imprinting, a general and effective approach for the preparation of hierarchically structured trimodal porous silica films was developed. With this new methodology, controlled formation of well-defined pore structures not only on macro- and mesoscale but also on microscale can be achieved, affording a new class of hierarchical porous materials with molecular recognition capability. As a demonstration, TNT was chosen as template molecule and hierarchically imprinted porous films were successfully fabricated, which show excellent sensing properties in terms of sensitivity, selectivity, stability, and regeneracy. The pore system reported here combines the multiple benefits arising from all length scales of pore size and simultaneously possesses a series of distinct properties such as high pore volume, large surface area, molecular selectivity, and rapid mass transport. Therefore, our described strategy and the resulting pore systems should hold great promise for various applications not only in chemical sensors, but also in catalysis, separation, adsorption, or electrode materials. PMID- 21644521 TI - Electrical conductivity enhancement of polymer/multiwalled carbon nanotube (MWCNT) composites by thermally-induced defunctionalization of MWCNTs. AB - We report a thermally-induced increase of electrical conductivity of polymer/multiwalled carbon nanotube (MWCNT) composites using Diels-Alder-adduct modified MWCNTs as additives. Thermal treatments of the composites induce the defunctionalization of the modified MWCNTs through retro-DA reaction, consequently to recover the electrical conductivity of MWCNTs and to increase the conductivity of PVDF/MWCNT composites. For the composites possessing 0.5 wt % of MWCNTs, thermal treatment increases the electrical conductivity from 2 * 10(-12) S cm(-1) to 4 * 10(-8) S cm(-1) and significantly reduces the value of percolation threshold. Meanwhile, the thermal treatment does not alter the mechanical properties of the composites. PMID- 21644522 TI - Water-mediated promotion of dye sensitization of TiO2 under visible light. AB - Preadsorbed water along with surrounding bulk water significantly modulates the surface electronic structure of TiO(2), switches the adsorption mode of dyes, and promotes dye sensitization of TiO(2) under visible-light irradiation. This opens a door toward facile improvement in the efficiency of photodegradation of dyes and dye-sensitized solar cells under visible-light irradiation without any complicated and expensive surface modulation. PMID- 21644523 TI - Low-temperature NMR studies on inosine wobble base pairs. AB - Base pairs formed by the inosine nucleoside (I) play an important role in many physiological processes as well as in various DNA technologies. Relative stabilities and favored base pair geometries of free inosine wobble base pairs in aprotic solvents have been determined through (1)H NMR measurements at room temperature and at very low temperatures in a freonic solvent. As indicated by its significantly deshielded imino proton, the Watson-Crick-type I.C base pair forms a remarkably strong NHN hydrogen bond. For the thermodynamically less stable I.A wobble base pair, two configurations of similar population coexist at 133 K in the slow hydrogen bond exchange regime, namely a Watson-Crick(I)-Watson Crick(A) geometry and a Watson-Crick(I)-Hoogsteen(A) geometry. I.U base pairs are stabilized by two rather weak hydrogen bonds and are significantly disfavored over inosine self-associates in a low-temperature Freon solution. PMID- 21644524 TI - Conversion of alpha-amino acids into bioactive o-aminoalkyl resorcylates and related dihydroxyisoindolinones. AB - The synthesis of biologically active o-aminoalkyl resorcylates and related dihydroxyisoindolinones from functionalized alpha-amino acids without the use of phenolic protection is described. The key aminoalkyl-diketo-dioxinone intermediates were prepared utilizing a crossed Claisen condensation reaction in the presence of diethylzinc. The aromatic unit was constructed via late stage cyclization and aromatization, and subsequent modification provided the novel resorcylates which showed activity against a selection of receptors and kinases, including 5-HT and CDK. PMID- 21644525 TI - A cooperative participation of the amido group in the organocatalytic construction of all-carbon quaternary stereocenters by Michael addition with beta ketoamides. AB - The secondary amido group of alpha-substituted beta-ketoamides plays a crucial role in the control of the reactivity and spatial arrangement (selectivity) in the organocatalyzed Michael addition to unsaturated carbonyls. This results in an unprecedented activation mode of substrates through H-bonding interactions allowing the construction of enantiomerically enriched functionalized all-carbon quaternary centers and spiroaminals of high synthetic potential. PMID- 21644526 TI - A "reactive" ratiometric fluorescent probe for mercury species. AB - A ratiometric fluorescent probe for mercury species is developed based on the metal-promoted hydrolysis of a vinyl ether derivative of 2-(benzothiazol-2 yl)phenol in a buffer solution. The probe responds selectively to mercury species over various other metal ions with a marked fluorescence change from blue to cyan through the excited state intramolecular proton transfer (ESIPT) process. The fluorescence titration is complete with 0.5 equiv of HgCl(2), which indicates that the probe also responds to organomercury species, RHgCl. PMID- 21644527 TI - Chemoselective Suzuki coupling of diborylmethane for facile synthesis of benzylboronates. AB - The chemoselective Pd-catalyzed Suzuki-Miyaura cross-coupling reaction using a diborylmethane is reported. The use of an equimolar amount of base with a diborylmethane realized chemoselective coupling for the synthesis of various benzylboronate derivatives. Sterically hindered aryl bromides can give products in moderate to excellent yields. PMID- 21644528 TI - Synthesis, structure, and reactivity of a mononuclear organozinc hydride complex: facile insertion of CO2 into a Zn-H Bond and CO2-promoted displacement of siloxide ligands. AB - Tris(2-pyridylthio)methane, [Tptm]H, has been employed to synthesize the mononuclear alkyl zinc hydride complex, [kappa(3)-Tptm]ZnH, which has been structurally characterized by X-ray diffraction. [kappa(3)-Tptm]ZnH provides access to a variety of other [Tptm]ZnX derivatives. For example, [kappa(3) Tptm]ZnH reacts with (i) R(3)SiOH (R = Me, Ph) to give [kappa(4)-Tptm]ZnOSiR(3), (ii) Me(3)SiX (X = Cl, Br, I) to give [kappa(4)-Tptm]ZnX, and (iii) CO(2) to give the formate complex, [kappa(4)-Tptm]ZnO(2)CH. The bis(trimethylsilyl)amide complex [kappa(3)-Tptm]ZnN(SiMe(3))(2) also reacts with CO(2), but the product obtained is the isocyanate complex, [kappa(4)-Tptm]ZnNCO. The formation of [kappa(4)-Tptm]ZnNCO is proposed to involve initial insertion of CO(2) into the Zn-N(SiMe(3))(2) bond, followed by migration of a trimethylsilyl group from nitrogen to oxygen to generate [kappa(4)-Tptm]ZnOSiMe(3) and Me(3)SiNCO, which subsequently undergo CO(2)-promoted metathesis to give [kappa(4)-Tptm]ZnNCO and (Me(3)SiO)(2)CO. PMID- 21644529 TI - N-fused imidazoles as novel anticancer agents that inhibit catalytic activity of topoisomerase IIalpha and induce apoptosis in G1/S phase. AB - On the basis of structures of known topoisomerase II catalytic inhibitors and initial molecular docking studies, bicyclic N-fused aminoimidazoles were predicted as potential topoisomerase II inhibitors. They were synthesized by multicomponent reactions and evaluated against human topoisomerase IIalpha (hTopoIIalpha) in decatenation, relaxation, cleavage complex, and DNA intercalation in vitro assays. Among 31 compounds of eight different bicyclic scaffolds, it was found that imidazopyridine, imidazopyrazole, and imidazopyrazine with suitable substituents exhibited potent inhibition of catalytic activity of hTopoIIalpha while not showing DNA intercalation. Molecular docking studies and molecular dynamics (MD) simulation analysis, ATPase-kinetics and ATP-dependent plasmid relaxation assay revealed the catalytic mode of inhibition of the title compounds plausibly by blocking the ATP-binding site. N Fused aminoimidazoles showed potent anticancer activities in kidney and breast cancer cell lines, low toxicity to normal cells, relatively higher potency compared to etoposide and 5-fluorouracil in kidney cancer cell lines, and potent inhibition in cell migration. These compounds were found to exert apoptotic effect in G1/S phase. PMID- 21644530 TI - Copper(II)-catalyzed room temperature aerobic oxidation of hydroxamic acids and hydrazides to acyl-nitroso and azo intermediates, and their Diels-Alder trapping. AB - CuCl(2), in the presence of a 2-ethyl-2-oxazoline ligand, is an effective catalyst for the room temperature, aerobic oxidation of hydroxamic acids and hydrazides, to acyl-nitroso and azo dienophiles respectively, which are efficiently trapped in situ via both inter- and intramolecular hetero-Diels-Alder reactions with dienes. Both inter- and intramolecular variants of the Diels-Alder reaction are suitable under the reaction conditions using a variety of solvents. Under the same conditions, an acyl hydrazide was also oxidized to give an acyl azo dienophile which was trapped intramolecularly by a diene. PMID- 21644531 TI - Palladium-catalyzed insertion of CO2 into vinylaziridines: new route to 5 vinyloxazolidinones. AB - 2-Vinylaziridines undergo a mild Pd-catalyzed ring-opening cyclization reaction with an ambient atmosphere of carbon dioxide to give 5-vinyloxazolidinones. The process is high yielding as well as regio- and stereoselective. PMID- 21644532 TI - Consecutive condensation, C-N and N-N bond formations: a copper- catalyzed one pot three-component synthesis of 2H-indazole. AB - 2H-Indazoles are synthesized using copper-catalyzed, one-pot, three-component reactions of 2-bromobenzaldehydes, primary amines, and sodium azide. A copper catalyst plays the key role in the formation of C-N and N-N bonds. This method has a broad substrate scope with a high tolerance for a variety of functional groups. PMID- 21644533 TI - Solution self-assembly of the sophorolipid biosurfactant and its mixture with anionic surfactant sodium dodecyl benzene sulfonate. AB - The self-assembly in aqueous solution of the acidic (AS) and lactonic (LS) forms of the sophorolipid biosurfactant, their mixtures, and their mixtures with anionic surfactant sodium dodecyl benzene sulfonate, LAS, has been studied using predominantly small-angle neutron scattering, SANS, at relatively low surfactant concentrations of <30 mM. The more hydrophobic lactonic sophorolipid forms small unilamellar vesicles at low surfactant concentrations, in the concentration range of 0.2 to 3 mM, and transforms via a larger unilamellar vesicle structure at 7 mM to a disordered dilute phase of tubules at higher concentrations, 10 to 30 mM. In marked contrast, the acidic sophorolipid is predominantly in the form of small globular micelles in the concentration range of 0.5 to 30 mM, with a lower concentration of larger, more planar aggregates (lamellar or vesicular) in coexistence. In mixtures of AS and LS, over the same concentration range, the micellar structure associated with the AS sophorolipid dominates the mixed-phase behavior. In mixtures of anionic surfactant LAS with the AS sophorolipid, the globular micellar structure dominates over the entire composition and concentration range studied. In contrast, mixtures of LAS with the LS sophorolipid exhibit a rich evolution in phase behavior with solution composition and concentration. At low surfactant concentrations, the small unilamellar vesicle structure present for LS-rich solution compositions evolves into a globular micelle structure as the solution becomes richer in LAS. At higher surfactant concentrations, the disordered lamellar structure present for LS-rich compositions transforms to small vesicle/lamellar coexistence, to lamellar/micellar coexistence, to micellar/lamellar coexistence, and ultimately to a pure micellar phase as the solution becomes richer in LAS. The AS sophorolipid surfactant exhibits self-assembly properties similar to those of most other weakly ionic or nonionic surfactants that have relatively large headgroups. However, the more hydrophobic nature of the lactonic sophorolipid results in a more complex and unusual evolution in phase behavior with concentration and with concentration and composition when mixed with anionic surfactant LAS. PMID- 21644534 TI - High-pressure synthesis and superconductivity of a new binary barium germanide BaGe3. AB - A new binary barium germanide BaGe(3) was prepared by high-pressure and high temperature reactions using a Kawai type multi-anvil press. It crystallizes in a hexagonal unit cell with a = 6.814(1) A, c = 5.027(8) A, and V = 202.2(5) A(3) (the space group P6(3)/mmc, No. 194). The unit cell contains two layers along the c axis composed of Ba atoms and Ge(3) triangular units. The triangular units stack along the c axis to form 1D columns in which the adjacent Ge(3) units turn to opposite directions. The columns, therefore, can be described as the face sharing stacking of elongated Ge(6) octahedra. Each Ba atom is surrounded by six columns. BaGe(3) is metallic and shows superconductivity at 4.0 K. The band structure calculations revealed that there are four conduction bands mainly composed of Ge 4p and Ba 5d orbitals. From Fermi surface analysis, we confirmed that three of them have a large contribution of Ge 4pz orbitals in the vicinity of the Fermi level and show a simple 1D appearance. The remaining one contains Ge 4px, 4py, and Ba 5d contributions and shows a 2D property. PMID- 21644535 TI - Controlled growth of self-organized hexagonal arrays of metallic nanorods using template-assisted glancing angle deposition for superhydrophobic applications. AB - The fabrication of controlled, self-organized, highly ordered tungsten and aluminum nanorods was accomplished via the aluminum lattice template-assisted glancing angle sputtering technique. The typical growth mechanism of traditional glancing angle deposition technique was biased by self-organized aluminum lattice seeds resulting in superior quality nanorods in terms of size control, distribution, and long range order. The morphology, size, and distribution of the nanorods were highly controlled by the characteristics of the template seeds indicating the ability to obtain metallic nanorods with tunable distributions and morphologies that can be grown to suit a particular application. Water wettability of hexagonally arranged tungsten and aluminum nanorods was studied after modifying their surface with 5 nm of Teflon AF 2400, as an example, to exhibit the significance of such a controlled growth of metallic nanorods. This facile and scalable approach to generate nano seeds to guide GLAD, with nano seeds fabricated by anodic oxidization of aluminum followed by chemical etching, for the growth of highly ordered nanorods could have significant impact in a wide range of applications such as anti-icing coating, sensors, super capacitors, and solar cells. PMID- 21644536 TI - Pressure-induced local structure distortions in Cu(pyz)F2(H2O)2. AB - We employed infrared spectroscopy along with complementary lattice dynamics and spin density calculations to investigate pressure-driven local structure distortions in the copper coordination polymer Cu(pyz)F(2)(H(2)O)(2). Here, pyz is pyrazine. Our study reveals rich and fully reversible local lattice distortions that buckle the pyrazine ring, disrupt the bc-plane O-H...F hydrogen bonding network, and reinforce magnetic property switching. The resiliency of the soft organic ring is a major factor in the stability of this material. Interestingly, the collective character of the lattice vibrations masks direct information on the Cu-N and Cu-O linkages through the series of pressure-induced Jahn-Teller axis switching transitions, although Cu-F bond softening is clearly identified above 3 GPa. These findings illustrate the importance of combined bulk and local probe techniques for microscopic structure determination in complex materials. PMID- 21644537 TI - Can a stepwise steady flow computational fluid dynamics model reproduce unsteady particulate matter separation for common unit operations? AB - Computational fluid dynamics (CFD) is emerging as a model for resolving the fate of particulate matter (PM) by unit operations subject to rainfall-runoff loadings. However, compared to steady flow CFD models, there are greater computational requirements for unsteady hydrodynamics and PM loading models. Therefore this study examines if integrating a stepwise steady flow CFD model can reproduce PM separation by common unit operations loaded by unsteady flow and PM loadings, thereby reducing computational effort. Utilizing monitored unit operation data from unsteady events as a metric, this study compares the two CFD modeling approaches for a hydrodynamic separator (HS), a primary clarifier (PC) tank, and a volumetric clarifying filtration system (VCF). Results indicate that while unsteady CFD models reproduce PM separation of each unit operation, stepwise steady CFD models result in significant deviation for HS and PC models as compared to monitored data; overestimating the physical size requirements of each unit required to reproduce monitored PM separation results. In contrast, the stepwise steady flow approach reproduces PM separation by the VCF, a combined gravitational sedimentation and media filtration unit operation that provides attenuation of turbulent energy and flow velocity. PMID- 21644538 TI - Trophic magnification of poly- and perfluorinated compounds in a subtropical food web. AB - Perfluorinated compounds (PFCs) are known to biomagnify in temperate and Arctic food webs, but little is known about their behavior in subtropical systems. The environmental distribution and biomagnification of PFCs, extractable organic fluorine (EOF), and total fluorine were investigated in a subtropical food web. Surface water, sediment, phytoplankton, zooplankton, gastropods, worms, shrimps, fishes, and waterbirds collected in the Mai Po Marshes Nature Reserve in Hong Kong were analyzed. Trophic magnification was observed for perfluorooctanesulfonate (PFOS), perfluorodecanoate (PFDA), perfluoroundecanoate (PFUnDA), and perfluorododecanoate (PFDoDA) in this food web. Risk assessment results for PFOS, PFDA, and perfluorooctanoate (PFOA) suggest that current PFC concentrations in waterbird livers are unlikely to pose adverse biological effects to waterbirds. All hazard ratio (HR) values reported for PFOS and PFOA are less than one, which suggests that the detected levels will not cause any immediate health effects to the Hong Kong population through the consumption of shrimps and fishes. However, only 10-12% of the EOF in the shrimp samples was comprised of known PFCs, indicating the need for further investigation to identify unknown fluorinated compounds in wildlife. PMID- 21644539 TI - Low packing density self-assembled superstructure of octahedral Pt3Ni nanocrystals. AB - We present a structural study of Pt(3)Ni nanoctahedron superlattice, prepared through both drop-casting and controlled solvent evaporation approaches. In this superlattice system containing ~10.6 nm side-length Pt(3)Ni nanoctahedra, we observed a body-centered cubic (bcc) packing structure in both local superlattices and statistically averaged superlattice ensembles using transmission electron microscopic tomography and grazing-incidence small-angle X ray scattering techniques, respectively. Within the superstructure, it was directly observed that nanoctahedra are orientated along the superstructure axes through sharing their vertices. We found that this arrangement of a bcc superstructure with nanoctahedra connecting through their vertices is dependent on neither the processing pathway nor the substrate under our experimental conditions. With such a very low packing density and ultrahigh surface area, this type of self-organized superstructure possesses unique features for future applications. PMID- 21644540 TI - Using fluorine nuclear magnetic resonance to probe changes in the structure and dynamics of membrane-active peptides interacting with lipid bilayers. AB - The antimicrobial peptide MSI-78 serves as a model system for studying interactions of bioactive peptides with membranes. Using a series of MSI-78 peptides that incorporate l-4,4,4-trifluoroethylglycine, a small and sensitive (19)F nuclear magnetic resonance probe, we investigated how the local structure and dynamics of the peptide change when it binds to the lipid bilayer. The fluorinated MSI-78 analogues exhibited position-specific changes in (19)F chemical shift ranging from 1.28 to -1.35 ppm upon binding to lipid bicelles. The largest upfield shifts are associated with the most hydrophobic positions in the peptide. Changes in solvent isotope effects (H(2)O/D(2)O) on (19)F chemical shifts were observed for the peptides that are consistent with the MSI-78 solvent inaccessible hydrophobic core upon binding bicelles. Transverse relaxation measurements of the (19)F nucleus, using the Carr-Purcell-Meiboom-Gill pulse sequence, were used to examine changes in the local mobility of MSI-78 that occur upon binding to the lipid bilayer. Positions in the hydrophobic core of peptide membrane complex show the greatest decrease in mobility upon binding of the lipid bilayer, whereas residues that interact with lipid headgroups are more mobile. The most mobile positions are at the N- and C-termini of the peptide. These results provide support for the proposed mechanism of membrane disruption by MSI 78 and reveal new details about the dynamic changes that accompany membrane binding. PMID- 21644541 TI - Antimalarial pyrido[1,2-a]benzimidazoles. AB - A novel class of antimalarial pyrido[1,2-a]benzimidazoles were synthesized and evaluated for antiplasmodial activity and cytotoxicity following hits identified from screening commercially available compound collections. The most active of these, TDR86919 (4c), showed improved in vitro activity vs the drug-resistant K1 strain of Plasmodium falciparum relative to chloroquine (IC(50) = 0.047 MUM v 0.17 MUM); potency was retained against a range of drug-sensitive and drug resistant strains, with negligible cytotoxicity against the mammalian (L-6) cell line (selectivity index of >600). 4c and several close analogues (as HCl or mesylate salts) showed significant efficacy in P. berghei infected mice following both intraperitoneal (ip) and oral (po) administration, with >90% inhibition of parasitemia, accompanied by an increase in the mean survival time (MSD). The pyrido[1,2-a]benzimidazoles appeared to be relatively slow acting in vivo compared to chloroquine, and metabolic stability of the alkylamino side chain was identified as a key issue in influencing in vivo activity. PMID- 21644543 TI - "Soft Si": effective stiffness of supported crystalline nanomembranes. AB - We investigate the effective mechanical response of a layered system consisting of a thin crystalline sheet (nanomembrane) on a bulk substrate, with a high elastic mismatch (in the range of 5 to 9 orders of magnitude) between the stiff sheet and the compliant substrate. Using finite-element mechanics models and indentation experiments ranging from micro to nano, we show that the mismatch between the sheet and substrate elastic moduli, the length scale of deformation, and the sheet thickness all play a significant role in defining the effective stiffness of the layered system. For a wide range of indenter sizes, the mechanical response of the composite system is indistinguishable from that of the compliant substrate. In particular, at large indenter sizes, the mechanical response of the layered system is dominated by that of the compliant substrate. For decreasing indenter sizes, the effective stiffness of the layered structure reaches a finite value different from either the one expected for the compliant substrate or for a bulk crystal of the same material as the stiff top membrane. PMID- 21644542 TI - Broad distribution of energetically important contacts across an extended protein interface. AB - Infection of cells by HIV depends upon profound structural rearrangements within the trimeric viral protein gp41. Critical to this process is the formation of a six-helix bundle in which a set of three N-terminal heptad repeat (NHR) helices assemble to form a core displaying long grooves that provide docking sites for three C-terminal heptad repeat (CHR) helices. We report experiments designed to discriminate between two alternative hypotheses regarding the source of affinity between individual CHR helices and the complementary groove: (1) affinity is dominated by interactions of a small cluster of side chains at one end of the CHR helix; or (2) affinity depends upon interactions distributed across the long CHR helix. We have employed two complementary experimental designs, and results from both favor the latter hypothesis. PMID- 21644544 TI - Cross-linked nanoassemblies from poly(ethylene glycol)-poly(aspartate) block copolymers as stable supramolecular templates for particulate drug delivery. AB - Block copolymer cross-linked nanoassemblies (CNAs) were developed as stable supramolecular templates for particulate drug delivery. Poly(ethylene glycol) poly(aspartate) [PEG-p(Asp)] block copolymers, consisting of PEG (5 or 12 kDa) and Asp (5, 14, 25, 33, and 37 repeating units), were used as scaffolds and grafts in combination to prepare a nanoassembly library of grafted nanoassemblies (GNAs) and CNAs. Four synthesis routes were tested to maximize the number of drug binding Asp units per nanoassembly. Grafting-onto-scaffold and grafting-from scaffold methods were used for GNA synthesis. Either partially or completely deprotected PEG-p(Asp) was cross-linked with diamine compounds to prepare CNAs. (1)H NMR and GPC measurements showed that GNAs and CNAs contained the maximum 183 and 253 Asp units, respectively. Initial screening of the nanoassemblies revealed that GNAs would be impractical for further development as drug carriers due to variable grafting efficiency and low product yields. CNAs were obtained in high yields and identified as a promising supramolecular template that can entrap and release ionizable drugs (doxorubicin), enhancing the particle stability of nanoassemblies in the pharmaceutically relevant pH ranges between 4 and 9. Light scattering measurements demonstrated that the particle size of CNAs remained uniform before and after drug entrapment, causing neither aggregation nor dissociation (<5 mg/mL). PMID- 21644545 TI - Ruthenium-catalyzed direct C-H bond arylations of heteroarenes. AB - Ruthenium-catalyzed C-H bond arylations of indoles, thiophenes, and pyrroles were accomplished in a highly chemo- and site-selective manner through the use of removable directing groups. PMID- 21644546 TI - Evaluation of several two-step scoring functions based on linear interaction energy, effective ligand size, and empirical pair potentials for prediction of protein-ligand binding geometry and free energy. AB - The performances of several two-step scoring approaches for molecular docking were assessed for their ability to predict binding geometries and free energies. Two new scoring functions designed for "step 2 discrimination" were proposed and compared to our CHARMM implementation of the linear interaction energy (LIE) approach using the Generalized-Born with Molecular Volume (GBMV) implicit solvation model. A scoring function S1 was proposed by considering only "interacting" ligand atoms as the "effective size" of the ligand and extended to an empirical regression-based pair potential S2. The S1 and S2 scoring schemes were trained and 5-fold cross-validated on a diverse set of 259 protein-ligand complexes from the Ligand Protein Database (LPDB). The regression-based parameters for S1 and S2 also demonstrated reasonable transferability in the CSARdock 2010 benchmark using a new data set (NRC HiQ) of diverse protein-ligand complexes. The ability of the scoring functions to accurately predict ligand geometry was evaluated by calculating the discriminative power (DP) of the scoring functions to identify native poses. The parameters for the LIE scoring function with the optimal discriminative power (DP) for geometry (step 1 discrimination) were found to be very similar to the best-fit parameters for binding free energy over a large number of protein-ligand complexes (step 2 discrimination). Reasonable performance of the scoring functions in enrichment of active compounds in four different protein target classes established that the parameters for S1 and S2 provided reasonable accuracy and transferability. Additional analysis was performed to definitively separate scoring function performance from molecular weight effects. This analysis included the prediction of ligand binding efficiencies for a subset of the CSARdock NRC HiQ data set where the number of ligand heavy atoms ranged from 17 to 35. This range of ligand heavy atoms is where improved accuracy of predicted ligand efficiencies is most relevant to real-world drug design efforts. PMID- 21644547 TI - Comparison of sessile drop and captive bubble methods on rough homogeneous surfaces: a numerical study. AB - Quasi-static experiments using sessile drops and captive bubbles are the most employed methods for measuring advancing and receding contact angles on real surfaces. These observable contact angles are the most easily accessible and reproducible. However, some properties of practical surfaces induce certain phenomena that cause a built-in uncertainty in the estimation of advancing and receding contact angles. These phenomena are well known in surface thermodynamics as stick-slip phenomena. Following the work of Marmur (Marmur, A. Colloids Surf., A 1998, 136, 209-215), where the stick-slip effects were studied with regard to sessile drops and captive bubbles on heterogeneous surfaces, we developed a novel extension of this study by adding the effects of roughness to both methods for contact angle measurement. We found that the symmetry between the surface roughness problem and the chemical heterogeneity problem breaks down for drops and bubbles subjected to stick-slip effects. PMID- 21644548 TI - New candidaspongiolides, tedanolide analogues that selectively inhibit melanoma cell growth. AB - Extracts of the sponge genus Candidaspongia showed selective cytotoxicity toward melanoma cells in the NCI 60-cell-line screen. Continued investigation of the Candidaspongia sp. extracts led to the isolation of three new tedanolide analogues, precandidaspongiolides A (1) and B (2) and candidaspongiolide B (4), as well as candidaspongiolide A (3) and tedanolide (5). Semisynthetic derivatives were also generated to develop SAR. Candidaspongiolides A/B were the most potent and showed low nanomolar activity against several melanoma cell lines. PMID- 21644549 TI - Self-assembling imidazolium-based ionic liquid in rigid nanopores induces anomalous CO2 adsorption at low pressure. AB - An alkylimidazolium-based long-chain ionic liquid (LCIL) was immobilized in silica nanopores via a supramolecular assembly approach. To discuss the characteristic features of LCIL in a confined nanospace, except for the characteristics of the host materials, we have prepared the silica host with monodisperse morphology and a nanostructured system to immobilize LCIL. The nanostructure is composed of three distinct regions: the silica framework, the hydrophobic interior of the alkyl chains, and the organic-inorganic ionic interface. Anomalous CO(2) adsorption sites were found to be well-ordered locations on the ionic interface fabricated by the pi-pi-stacked imidazolium heads containing inorganic anions and polar silica surfaces. PMID- 21644550 TI - How to make the Cassie wetting state stable? AB - Wetting of rough hydrophilic and hydrophobic surfaces is discussed. The stability of the Cassie state, with air trapped in relief details under the droplet, is necessary for the design of true superhydrophobic surfaces. The potential barrier separating the Cassie state and the Wenzel state, for which the substrate is completely wetted, is calculated for both hydrophobic and hydrophilic surfaces. When the surface is hydrophobic, the multiscaled roughness of pillars constituting the surface increases the potential barrier separating the Cassie and Wenzel states. When water fills the hydrophilic pore, the energy gain due to the wetting of the pore hydrophilic wall is overcompensated by the energy increase because of the growth of the high-energetic liquid-air interface. The potential barrier separating the Cassie and Wenzel states is calculated for various topographies of surfaces. Structural features of reliefs favoring enhanced hydrophobicity are elucidated. PMID- 21644551 TI - Combinatorial discovery of fluorescent pharmacophores by multicomponent reactions in droplet arrays. AB - Fluorescence imaging in clinical diagnostics and biomedical research relies to a great extent on the use of small organic fluorescent probes. Because of the difficulty of combining fluorescent and molecular-recognition properties, the development of such probes has been severely restricted to a number of well-known fluorescent scaffolds. Here we demonstrate that autofluorescing druglike molecules are a valuable source of bioimaging probes. Combinatorial synthesis and screening of chemical libraries in droplet microarrays allowed the identification of new types of fluorophores. Their concise and clean assembly by a multicomponent reaction presents a unique potential for the one-step synthesis of thousands of structurally diverse fluorescent molecules. Because they are based upon a druglike scaffold, these fluorophores retain their molecular recognition potential and can be used to design specific imaging probes. PMID- 21644552 TI - Creating a pseudometallic state of K+ by intercalation into 18-crown-6 grafted on polyfluorene as electron injection layer for high performance PLEDs with oxygen- and moisture-stable Al cathode. AB - Polymer light-emitting diodes (PLEDs) suffer from inadequate lifetimes because of the use of environmentally sensitive metals as the cathodes. We present the use of water/methanol-soluble polyfluorene grafted with 18-crown-6 chelating to K(+) as the electron-injection layer (EIL) for deep-blue-emission PLEDs, allowing the use of environmentally stable Al as the cathode since electron donation from the 18-crown-6 can reduce K(+) to a stable "pseudometallic state", enabling it to act as an intermediate step for electron injection. Furthermore, when poly(ethylene oxide) was blended into the EIL to provide hole blocking (HB), the device exhibited the highest performance reported to date for a deep-blue-emission PLED based on a conjugated polymer as the emitting layer, with a brightness of 54,800 cd/m(2) and an external quantum efficiency of 5.42%. The use of such an EI-HB layer opens a broad avenue leading toward industrialization of PLEDs. PMID- 21644553 TI - Improving dielectric properties of BaTiO3/ferroelectric polymer composites by employing surface hydroxylated BaTiO3 nanoparticles. AB - Dielectric properties of poly(vinylidene fluoride) (PVDF) based nanocomposites filled with surface hydroxylated BaTiO(3) (h-BT) nanoparticles were reported. The h-BT fillers were prepared from crude BaTiO(3) (c-BT) in aqueous solution of H(2)O(2). Results showed that the dielectric properties of the h-BT/PVDF nanocomposites had weaker temperature and frequency dependences than that of c BT/PVDF nanocomposites. Meanwhile, the h-BT/PVDF composites showed lower loss tangent and higher dielectric strength. It is suggested that the strong interaction between h-BT fillers and PVDF matrix is the main reason for the improved dielectric properties. PMID- 21644554 TI - Synthesis of functionalized 2-aminohydropyridines and 2-pyridinones via domino reactions of arylamines, methyl propiolate, aromatic aldehydes, and substituted acetonitriles. AB - An efficient and practical synthetic method for the functionalized 2-amino hydropyridines and 2-pyridinones was successfully developed via the domino reactions of arylamines, methyl propiolate, aromatic aldehydes and the substituted acetonitriles with triethylamine as base catalyst. Reactions involving malononitrile and cyanoacetamide gave exclusively the 2 aminohydropyridines. On the other hand ethyl cyanoacetate resulted in the 2 pyridinones as main products. PMID- 21644555 TI - Electrospray preparation of hierarchically-structured mesoporous TiO2 spheres for use in highly efficient dye-sensitized solar cells. AB - We report a simple method to prepare hierarchically structured TiO(2) spheres (HS TiO(2)), using an electrostatic spray technique, that are utilized for photoelectrodes of highly efficient dye-sensitized solar cells (DSSCs). This method has an advantage to remove the synthesis steps in conventional sol-gel method to form nano-sized spheres of TiO(2) nanoclusters. The fine dispersion of commercially available nanocrystalline TiO(2) particles (P25, Degussa) in EtOH without surfactants and additives is electro-sprayed directly onto a fluorine dopoed tin-oxide (FTO) substrate for DSSC photoelectrodes. The DSSCs of HS-TiO(2) photoelectrodes show high energy conversion efficiency over 10% under illumination of light at 100 mW cm(-2), AM1.5 global. It is concluded from frequency-dependent measurements that the faster electron diffusion coefficient and longer lifetime of HS-TiO(2) than those in nonstructured TiO(2) contribute to the enhanced efficiency in DSSCs. PMID- 21644556 TI - Conformal polymeric thin films by low-temperature rapid initiated chemical vapor deposition (iCVD) using tert-butyl peroxybenzoate as an initiator. AB - Conformal poly(cyclohexyl methacrylate) (pCHMA) thin films were synthesized via initiated chemical vapor deposition (iCVD), with tert-butyl peroxybenzoate (TBPOB) as the initiator, representing the first time that TBPOB has been used as an initiator for iCVD synthesis. Using TBPOB instead of tert-butyl peroxide (TBPO), the rate of iCVD film growth increased by a factor of up to seven at comparable conformality and lower the filament temperature from 257 to 170 degrees C at a comparable deposition rate of 3 nm/min. The conformal deposition of functional thin films is desired for applications including microfluidics, medical devices and membranes. Lower filament temperatures reduce the heat load to the deposition surface and thus are advantageous for polymeric substrates that are temperature sensitive or monomers that decompose at high temperatures. Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS) results demonstrate the similarity of the TBPOB- to the TBPO initiated pCHMA main chains. However, the aromatic group in TBPOB provided a unique spectral signature of the polymer chain end group in the FTIR and the peak intensity increased with increase of filament temperature. Scanning electron micrographs (SEMs) revealed that the pCHMA coatings are conformal over non-planar structures; however, at identical process conditions, TBPO-initiated films showed a slightly better conformality due to the lower sticking coefficient of TBPO. At a monomer partial pressure of 0.45, TBPOB has a sticking coefficient value of 0.1188 +/- 0.0092, which is ~3 times as high as that of TBPO (0.0413 +/- 0.0058). The step coverage is insensitive to filament temperature if the surface concentration of the monomer is fixed. PMID- 21644557 TI - High-throughput combinatorial database of electronic band structures for inorganic scintillator materials. AB - For the purpose of creating a database of electronic structures of all the known inorganic compounds, we have developed a computational framework based on high throughput ab initio calculations (AFLOW) and an online repository (www.aflowlib.org). In this article, we report the first step of this task: the calculation of band structures for 7439 compounds intended for the research of scintillator materials for gamma-ray radiation detection. Data-mining is performed to select the candidates from 193,456 compounds compiled in the Inorganic Crystal Structure Database. Light yield and scintillation nonproportionality are predicted based on semiempirical band gaps and effective masses. We present a list of materials, potentially bright and proportional, and focus on those exhibiting small effective masses and effective mass ratios. PMID- 21644558 TI - [H(x)TeV9O28]((5-x)-) (x=1 and 2): vanadotellurates with decavanadate structure. AB - Two new vanadotellurates, [HTeV(9)O(28)](4-) and [H(2)TeV(9)O(28)](3-) have been synthesized and structurally characterized as tetra-n-butylammonium (TBA) salts: TBA(4)[HTeV(9)O(28)].2CH(3)CN [triclinic, space group P 1, a = 16.7102(6) A, b = 17.4680(7) A, c = 17.9634(7) A, alpha = 74.412(1) degrees , beta = 67.494(1) degrees , gamma = 74.160(2) degrees , Z = 2] and TBA(3)[H(2)TeV(9)O(28)] [monoclinic, space group P2(1)/c, a = 13.0013(5) A, b = 19.157(1) A, c = 28.453(1) A, beta = 97.222(2) degrees , Z = 4]. The results of the structural analyses indicate that the four O atoms that bridge two V atoms on the Te side are the most basic ones in the structure. The results of density-functional theory (DFT) calculations support this view. PMID- 21644559 TI - Adsorption, desorption, and thermodynamic studies of CO2 with high-amine-loaded multiwalled carbon nanotubes. AB - Commercially available multiwalled carbon nanotubes (CNTs) were functionalized with a high mass load of 3-aminopropyltriethoxysilane (APTS) to study their behaviors in the cyclic CO(2) adsorption as well as the associated thermodynamic properties. The breakthrough curve showed a fast kinetics of CO(2) adsorption resulting in percentage ratios of working capacity to equilibrium capacity greater than 80%. The adsorption capacity of CNT(APTS) was significantly influenced by the presence of water vapor and reached a maximum of 2.45 mmol/g at a water vapor of 2.2%. The adsorption capacities and the physicochemical properties of CNT(APTS) were preserved through 100 adsorption-desorption cycles displaying the stability of CNT(APTS) during a prolonged cyclic operation. The heat input required to regenerate spent CNT(APTS) was determined, and the result suggests that adsorption process with solid CNT(APTS) is possibly a promising CO(2) capture technology. PMID- 21644560 TI - nPEG-TiO2 nanoparticles: a facile route to elaborate nanostructured surfaces for biological applications. AB - We report the synthesis of diacid-terminated PEG-functionalized cubic TiO(2) nanocrystals by a simple one-step solvothermal method, and their further use to form nanostructured surfaces for protein immobilization. The relevance and major interest of the so-obtained nanocrystals are the presence of terminal carboxylic acid groups at their surface, as confirmed by infrared analyses, in addition to the surrounding PEG chains, essential to avoid non specific interactions. These functional chemical groups were used to (i) immobilize the synthesized nanocubes on a cysteamine-modified Au surface, and to (ii) attach proteins via a presumable covalent link. AFM images show that the shapes and the narrow size distribution of the nanocubes, observed by TEM, were preserved after their immobilization on the modified Au surface. Moreover, the efficiency and specificity of antigen recognition were demonstrated using spectroscopic analyses. Our successful approach provides a versatile and facile way to elaborate specific and sensitive nanostructured surfaces for biosensors. PMID- 21644561 TI - Photocycloaddition of biscyclopropyl alkenes to C60: an unprecedented approach toward cis-1 tricyclic-fused fullerenes. AB - A novel, simple, and entirely regioselective tandem cycloaddition of biscyclopropyl-substituted alkenes to [60]fullerene has been revealed. This reaction affords cis-1 tricyclic-fused organofullerenes bearing the hitherto elusive 5-4-5 fused tricyclic ring system. PMID- 21644562 TI - Combinatorial and high-throughput screening of materials libraries: review of state of the art. AB - Rational materials design based on prior knowledge is attractive because it promises to avoid time-consuming synthesis and testing of numerous materials candidates. However with the increase of complexity of materials, the scientific ability for the rational materials design becomes progressively limited. As a result of this complexity, combinatorial and high-throughput (CHT) experimentation in materials science has been recognized as a new scientific approach to generate new knowledge. This review demonstrates the broad applicability of CHT experimentation technologies in discovery and optimization of new materials. We discuss general principles of CHT materials screening, followed by the detailed discussion of high-throughput materials characterization approaches, advances in data analysis/mining, and new materials developments facilitated by CHT experimentation. We critically analyze results of materials development in the areas most impacted by the CHT approaches, such as catalysis, electronic and functional materials, polymer-based industrial coatings, sensing materials, and biomaterials. PMID- 21644563 TI - Diversity-oriented synthesis of 2,4,6-trisubstituted piperidines via type II anion relay chemistry. AB - An effective, general protocol for the Diversity-Oriented Synthesis (DOS) of 2,4,6-trisubstituted piperidine congeners has been designed and validated. The successful strategy entails a modular approach to all possible stereoisomers of the selected piperidine scaffold, exploiting Type II Anion Relay Chemistry (ARC), followed in turn by intramolecular S(N)2 cyclization, chemoselective removal of the dithiane moieties and carbonyl reductions. PMID- 21644564 TI - Synthesis of parvistemin A via biomimetic oxidative dimerization. AB - The first synthesis of the naturally occurring benzoquinone dimer parvistemin A is reported. The key step is the late stage iron(III) mediated dimerization of a 1,2,4-trihydroxyarene to give the natural product in good yield, a phenol oxidative coupling that is believed to be biomimetic. The route proceeds in seven steps from an inexpensive commercially available acetophenone in 14% overall yield. PMID- 21644565 TI - Efficient tuning of LUMO levels of 2,5,8,11-substituted perylenediimides via copper catalyzed reactions. AB - Via one-step copper catalyzed procedures it was possible to synthesize 2,5,8,11 tetrabromo, tetrachloro, and tetracyano derivatives of perylenediimides. Characterization of optical and electrochemical properties of these materials proves substantial enhancement of the electron affinity, with a LUMO level as low as -4.4 eV in the case of the tetracyano perylenediimide. PMID- 21644566 TI - Impact of micelle ionic electrical double layer structure on the excited state protolytic reaction in the fluorescent probe bound to the colloidal nanoparticles. AB - The fluorescent probe, 2-hydroxynaphthalene(dodecylo)-6-sulfonamide (NSDA) bound selectively to shear plane of various electrostatic charges was synthesized and its photophysical properties have been investigated by means of steady state fluorescence and nanosecond time-resolved spectroscopy. Our experimental data allowed us to determine the excited state proton transfer (ESPT) rate and equilibrium constants of NSDA bound to micelles and to estimate the electric potential value (Psi) at the particle surface. The spatial dependence of proton movement velocity through electric double layer (EDL) of micelles has been thoroughly analyzed. In this article, a new approach of estimating the values of the micelle potential (Psi(R)) from the excited state proton transfer rate constant of the fluorescent probe bound at a certain distance (R) to a micellar surface has been proposed. The Psi(R) values, obtained in this way, are compared with electrophoretic values of the particle potential (zeta). Our results on electrophoretic potentials and the reaction course of the ESPT in colloidal environment may contribute to a deeper understanding of micellar interactions and behavior of the living cells in contact with various diluted substances such as pharmacological drugs, hormones, proteins, and other colloidal particles. PMID- 21644567 TI - n-InAs nanopyramids fully integrated into silicon. AB - InAs with an extremely high electron mobility (up to 40,000 cm(2)/V s) seems to be the most suitable candidate for better electronic devices performance. Here we present a synthesis of inverted crystalline InAs nanopyramids (NPs) in silicon using a combined hot ion implantation and millisecond flash lamp annealing techniques. Conventional selective etching was used to form the InAs/Si heterojunction. The current-voltage measurement confirms the heterojunction diode formation with the ideality factor of eta = 4.6. Kelvin probe force microscopy measurements indicate a type-II band alignment of n-type InAs NPs on p-type silicon. The main advantage of our method is its integration with large-scale silicon technology, which also allows applying it for Si-based electronic devices. PMID- 21644568 TI - Nickel(0)-catalyzed cyclization of N-benzoylaminals for isoindolinone synthesis. AB - A nickel(0) catalyst effectively mediates the cyclization of N-benzoyl aminals in the presence of a stoichiometric Lewis acid. This method enables preparation of a variety of isoindolinones with substitution on the benzoyl fragment and C-3 carbon. This reaction likely proceeds via an alpha-amidoalkylnickel(II) intermediate, which then may cyclize via either an electrophilic aromatic substitution or an insertion pathway. PMID- 21644569 TI - Effect of genipin crosslinking on the optical spectral properties and structures of collagen hydrogels. AB - Genipin, a natural cross-linking reagent extracted from the fruits of Gardenia jasminoides, can be effectively employed in tissue engineering applications due to its low cytotoxicity and high biocompatibility. The cross-linking of collagen hydrogels with genipin was followed with one-photon fluorescence spectroscopy, second harmonic generation, fluorescence and transmission electron microscopy. The incubation with genipin induced strong auto-fluorescence within the collagen hydrogels. The fluorescence emission maximum of the fluorescent adducts formed by genipin exhibit a strong dependence on the excitation wavelength. The emission maximum is at 630 nm when we excite the cross-linked samples with 590 nm light and shifts to 462 nm when we use 400 nm light instead. The fluorescence imaging studies show that genipin induces formation of long aggregated fluorescent strands throughout the depth of samples. The second harmonic generation (SHG) imaging studies suggest that genipin partially disaggregates 10 MUm "fiberlike" collagen structures because of the formation of these fluorescent cross-links. Transmission electron microscopy (TEM) studies reveal that genipin largely eliminates collagen's characteristic native fibrillar striations. Our study is the first one to nondestructively follow and identify the structure within collagen hydrogels in situ and to sample structures formed on both micro- and nanoscales. Our findings suggest that genipin cross-linking of collagen follows a complex mechanism and this compound modifies the structure within the collagen hydrogels in both micro- and nanoscale. PMID- 21644571 TI - Facile preparation of nitrogen-doped few-layer graphene via supercritical reaction. AB - To achieve the applications of graphene, the modulation of its electrical properties is of great significance. The element doping might give a promising approach to produce fascinating properties of graphene. Herein we report a facile chemical doping method to obtain nitrogen-doped (N-doped) few-layer graphene sheets through supercritical (SC) reaction in acetonitrile at temperature as low as 310 degrees C, using expanded graphite as starting material. X-ray photoelectron spectroscopy analysis revealed that the level of nitrogen-doping (N doping) increased from 1.57 to 4.56 at % when the reaction time was tuned from 2 to 24 h. Raman spectrum confirmed that the resulting N-doped few-layer graphene by SC reaction maintain high quality without any significant structural defects. Electrical measurements indicated that N-doped few-layer graphene sheets exhibit a typical n-type field-dependent behavior, suggesting the N-doping into the lattice of graphene. This work provides a convenient chemical route to the scalable production of N-doped graphene for potential applications in nanoelectronic devices. PMID- 21644570 TI - Imidazolopiperazines: hit to lead optimization of new antimalarial agents. AB - Starting from a hit series from a GNF compound library collection and based on a cell-based proliferation assay of Plasmodium falciparum, a novel imidazolopiperazine scaffold was optimized. SAR for this series of compounds is discussed, focusing on optimization of cellular potency against wild-type and drug resistant parasites and improvement of physiochemical and pharmacokinetic properties. The lead compounds in this series showed good potencies in vitro and decent oral exposure levels in vivo. In a Plasmodium berghei mouse infection model, one lead compound lowered the parasitemia level by 99.4% after administration of 100 mg/kg single oral dose and prolonged mice survival by an average of 17.0 days. The lead compounds were also well-tolerated in the preliminary in vitro toxicity studies and represents an interesting lead for drug development. PMID- 21644572 TI - Antibiofouling hybrid dendritic Boltorn/star PEG thiol-ene cross-linked networks. AB - A series of thiol-ene generated amphiphilic cross-linked networks was prepared by reaction of alkene-modified Boltorn polyesters (Boltorn-ene) with varying weight percent of 4-armed poly(ethylene glycol) (PEG) tetrathiol (0-25 wt%) and varying equivalents of pentaerythritol tetrakis(3-mercaptopropionate) (PETMP) (0-64 wt%). These materials were designed to present complex surface topographies and morphologies, with heterogeneity of surface composition and properties and robust mechanical properties, to serve as nontoxic antibiofouling coatings that are amenable to large-scale production for application in the marine environment. Therefore, a two-dimensional matrix of materials compositions was prepared to study the physical and mechanical properties, over which the compositions spanned from 0 to 25 wt% PEG tetrathiol and 0-64 wt% PETMP (the overall thiol/alkene (SH/ene) ratios ranged from 0.00 to 1.00 equiv), with both cross-linker weight percentages calculated with respect to the weight of Boltorn-ene. The Boltorn-ene components were prepared through the esterification of commercially available Boltorn H30 with 3-butenoic acid. The subsequent cross-linking of the Boltorn-PEG PETMP films was monitored using IR spectroscopy, where it was found that near complete consumption of both thiol and alkene groups occurred when the stoichiometry was ca. 48 wt% PETMP (0.75 equiv SH/ene, independent of PEG amount). The thermal properties of the films showed an increase in T(g) with an increase in 4-armed PEG-tetrathiol wt%, regardless of the PETMP concentration. Investigation of the bulk mechanical properties in dry and wet states found that the Young's modulus was the greatest at 48 wt% PETMP (0.75 equiv of SH/ene). The ultimate tensile strength increased when PETMP was constant and the PEG concentration was increased. The Young's modulus was slightly lower for wet films at constant PEG or constant PETMP amounts, than for the dry samples. The nanoscopic surface features were probed using atomic force microscopy (AFM), where it was observed that the surface of the amphiphilic films became increasingly rough with increasing PEG wt%. On the basis of the physicochemical data from the diverse sample matrix, a focused compositional profile was then investigated further to determine the antifouling performance of the cross-linked Boltorn-PEG-PETMP networks. For these studies, a low, constant PETMP concentration of 16 wt% was maintained with variation in the PEG wt% (0-35 wt%). Antifouling and fouling-release activities were tested against the marine alga Ulva. Spore settlement densities were low on these films, compared to that on standards of polydimethylsiloxane and glass. PMID- 21644573 TI - Microbial reverse electrodialysis cells for synergistically enhanced power production. AB - A new type of bioelectrochemical system for producing electrical power, called a microbial reverse-electrodialysis cell (MRC), was developed to increase voltages and power densities compared to those generated individually by microbial fuel cells (MFCs) or reverse electrodialysis (RED) systems. In RED systems, electrode overpotentials create significant energy losses due to thermodynamically unfavorable electrode reactions, and therefore a large number of stacked cells must be used to have significant energy recovery. This results in high capital costs for the large number of membranes, and increases energy losses from pumping water through a large number of cells. In an MRC, high overpotentials are avoided through oxidation of organic matter by exoelectrogenic bacteria on the anode and oxygen reduction on the cathode. An MRC containing only five pairs of RED cells, fed solutions typical of seawater (600 mM NaCl) and river water (12 mM NaCl) at 0.85 mL/min, produced up to 3.6 W/m(2) (cathode surface area) and 1.2-1.3 V with acetate as a substrate. Pumping accounted for <2% of the produced power. A higher flow rate (1.55 mL/min) increased power densities up to 4.3 W/m(2). COD removal was 98% with a Coulombic efficiency of 64%. Power production by the individual components was substantially lower with 0.7 W/m(2) without salinity driven energy, and <0.015 W/m(2) with reduced exoelectrogenic activity due to substrate depletion. These results show that the combination of an MFC and a RED stack synergistically increases performance relative to the individual systems, producing a new type of system that can be used to more efficiently capture salinity driven energy from seawater and river water. PMID- 21644574 TI - Polycarbonates from the polyhydroxy natural product quinic acid. AB - Strategies for the preparation of polycarbonates, derived from natural polyhydroxy monomeric repeat units, were developed for biosourced polycarbonates based on quinic acid. The design and synthesis of regioselectively tert butyldimethylsilyloxy (TBS)-protected 1,4- and 1,5-diol monomers of quinic acid were followed by optimization of their copolymerizations with phosgene, generated in situ from trichloromethyl chloroformate, to yield protected poly(1,4-quinic acid carbonate) and poly(1,5-quinic acid carbonate). The molecular weights reached ca. 7.6 kDa, corresponding to degrees of polymerization of ca. 24, with polydispersities ranging from 2.0 to 3.5, as measured by SEC using tetrahydrofuran as the eluent and with polystyrene calibration standards. Partially because of the presence of the bicyclic backbone, each regioisomeric poly(quinic acid carbonate) exhibited relatively high glass-transition temperatures, 209 degrees C for poly(1,4-quinic acid carbonate) and 229 degrees C for poly(1,5-quinic acid carbonate). Removal of the TBS-protecting groups was studied under mild conditions to achieve control over potential competing reactions involving polymer degradation, which could include cleavage of lactones within the repeat units, carbonate linkages, or both between the repeat units. Full deprotection was not achieved without some degree of polymer degradation. The regiochemistry of the monomer showed significant impact on the reactivity during deprotection and also on the thermal properties, with the 1,5 regioisomeric polymer having lower reactivity and giving higher T(g) values, in comparison with the 1,4-regioisomer. Each regioisomer underwent a 10-20 degrees C increase in T(g) upon partial removal of the TBS-protecting groups. As the extent of deprotection increased, the solubility decreased. Ultimately, at long deprotection reaction times, the solubility increased and the T(g) decreased because of significant degradation of the polymers. PMID- 21644575 TI - Multistimuli responsive micelles formed by a tetrathiafulvalene-functionalized amphiphile. AB - An electroactive tetrathiafulvalene (TTF)-functionalized amphiphile 1 was designed and synthesized to investigate its self-assembling behavior in water. Dynamic light scattering (DLS), (1)H NMR, fluorescence spectrum, and cryogenic transmission electron microscopy (cryo-TEM) studies revealed that amphiphile 1 can form micelle-like aggregates via direct dissolution into water, and the micellar architectures could be disrupted either by addition of chemical oxidant Fe(ClO(4))(3) or by complexation with electron-deficient cyclobis(paraquat-p phenylene) tetracation cyclophane (CBPQT(4+)) to release encapsulated hydrophobic dye Nile Red from the interior of micelles. PMID- 21644576 TI - Organogold reactivity with palladium, nickel, and rhodium: transmetalation, cross coupling, and dual catalysis. AB - Using two transition metals to simultaneously catalyze a reaction can offer distinct opportunities for reactivity and selectivity when compared to using single-metal catalyst systems. Creating dual transition metal catalytic systems is complicated, however, by challenges in predicting compatible reactivities and designing turnover pathways for both metals. In this Account, we describe our development of dual-metal catalysis reactions involving gold and a second transition metal. The unique rearrangement intermediates accessible through gold only catalysis, which exploits the soft Lewis acidity of Au(I), make gold an attractive partner for dual-metal catalysis reactions. Because of the complexity of achieving simultaneous turnover of two catalysts and predicting compatibilities, our approach has been to first gain a fundamental understanding of the reactivity of the two metals with each other, both in stoichiometric and monocatalyzed reactions. To this end, we have investigated the combined reactivity of organogold compounds with palladium, nickel, and rhodium. We narrate the intricacies of turning over two catalysts simultaneously and thereby illuminate the valuable role of fundamental studies in identifying the optimal conditions to promote desirable two-metal reactivity and compatibility. Transmetalation, redox reactivity, and new mechanisms for dual-metal catalytic turnover were probed from this standpoint. We have applied the knowledge gained through these studies to the development of reactions that are dual-catalyzed by gold and palladium, as well as nickel- and rhodium-catalyzed reactions of organogold compounds. More broadly, these new reactions expand the reactivity available to catalytic organogold intermediates via trapping and functionalization reactions with other transition metals. Our investigations reveal strategies useful for designing dual-metal reactions with gold. First, the versatility of gold as a transmetalation partner suggests that many potential methods may exist to intercept catalytic organogold intermediates with a second transition metal. Second, ligands on both metals should be selected carefully in order to prevent catalyst deactivation. Finally, reactions must be designed such that any oxidative steps involving the second metal outcompete undesired reactions with redox-active organogold compounds. We believe that the application of these principles will allow for the design of a diverse set of dual-catalyzed functionalizations befitting the wide variety of gold-catalyzed transformations already established. PMID- 21644577 TI - Structural changes in microcrystalline cellulose in subcritical water treatment. AB - Subcritical water is a high potential green chemical for the hydrolysis of cellulose. In this study microcrystalline cellulose was treated in subcritical water to study structural changes of the cellulose residues. The alterations in particle size and appearance were studied by scanning electron microscopy (SEM) and those in the degree of polymerization (DP) and molar mass distributions by gel permeation chromatography (GPC). Further, changes in crystallinity and crystallite dimensions were quantified by wide-angle X-ray scattering and (13)C solid-state NMR. The results showed that the crystallinity remained practically unchanged throughout the treatment, whereas the size of the remaining cellulose crystallites increased. Microcrystalline cellulose underwent significant depolymerization in subcritical water. However, depolymerization leveled off at a relatively high degree of polymerization. The molar mass distributions of the residues showed a bimodal form. We infer that cellulose gets dissolved in subcritical water only after extensive depolymerization. PMID- 21644578 TI - Environmental impacts of water use in global crop production: hotspots and trade offs with land use. AB - Global crop production is causing pressure on water and land resources in many places. In addition to local resource management, the related environmental impacts of commodities traded along international supply chains need to be considered and managed accordingly. For this purpose, we calculate the specific water consumption and land use for the production of 160 crops and crop groups, covering most harvested mass on global cropland. We quantify indicators for land and water scarcity with high geospatial resolution. This facilitates spatially explicit crop-specific resource management and regionalized life cycle assessment of processed products. The vast cultivation of irrigated wheat, rice, cotton, maize, and sugar cane, which are major sources of food, bioenergy, and fiber, drives worldwide water scarcity. According to globally averaged production, substituting biofuel for crude oil would have a lower impact on water resources than substituting cotton for polyester. For some crops, water scarcity impacts are inversely related to land resource stress, illustrating that water consumption is often at odds with land use. On global average, maize performs better than rice and wheat in the combined land/water assessment. High spatial variability of water and land use related impacts underlines the importance of appropriate site selection for agricultural activities. PMID- 21644579 TI - Synthesis and optical properties of acidochromic amine-substituted benzo[a]phenazines. AB - A new series of alkylamine- or arylamine-substituted benzo[a]phenazines have been synthesized from 1,2-naphthoquinones by employing simple sequential Michael-type addition with a variety of primary and secondary amines and the condensation reaction of the resulting amine-substituted 1,2-naphthoquinones with o phenylenediamine. They exhibited absorption peaks originating from the charge transfer transition between the amine and pyrazine segments and benzo[a]phenazine localized pi-pi* transitions. Although the absorption spectra of the dyes were not significantly influenced by the nature of the solvents, addition of TFA led to a prominent red-shift in the absorption spectra owing to the protonation at the quinoxaline segment which enhanced the electron-accepting ability. The qualitative trends observed in the optical properties and acidochromism were supported by density functional theoretical computations. The dyes displayed positive solvatochromism in the emission spectra suggestive of a more polar excited state. The dyes were also characterized by a quasi-reversible reduction couple originating from the pyrazine segment which underwent shifts corresponding to electron-donating strength of the amine segment. PMID- 21644580 TI - Negative polaron and triplet exciton diffusion in organometallic "molecular wires". AB - The dynamics of negative polaron and triplet exciton transport within a series of monodisperse platinum (Pt) acetylide oligomers is reported. The oligomers consist of Pt-acetylide repeats, [PtL(2)-C=C-Ph-C=C-](n) (where L = PBu(3) and Ph = 1,4 phenylene, n = 2, 3, 6, and 10), capped with naphthalene diimide (NDI) end groups. The Pt-acetylide segments are electro- and photoactive, and they serve as conduits for transport of electrons (negative polaron) and triplet excitons. The NDI end groups are relatively strong acceptors, serving as traps for the carriers. Negative polaron transport is studied by using pulse radiolysis/transient absorption at the Brookhaven National Laboratory Laser Electron Accelerator Facility (LEAF). Electrons are rapidly attached to the oligomers, with some fraction initially residing upon the Pt-acetylide chains. The dynamics of transport are resolved by monitoring the spectral changes associated with transfer of electrons from the chain to the NDI end group. Triplet exciton transport is studied by femtosecond-picosecond transient absorption spectroscopy. Near-UV excitation leads to rapid production of triplet excitons localized on the Pt-acetylide chains. The excitons transport to the chain ends, where they are annihilated by charge separation with the NDI end group. The dynamics of triplet transport are resolved by transient absorption spectroscopy, taking advantage of the changes in spectra associated with decay of the triplet exciton and rise of the charge-separated state. The results indicate that negative polarons and excitons are transported rapidly, on average moving distances of ~3 nm in less than 200 ps. Analysis of the dynamics suggests diffusive transport by a site-to-site hopping mechanism with hopping times of ~27 ps for triplets and <10 ps for electrons. PMID- 21644581 TI - Phase behavior of rod-coil diblock copolymer and homopolymer blends from self consistent field theory. AB - The phase behavior of binary blends of rod-coil diblock copolymers and coil or rod homopolymers is studied by the self-consistent field theory (SCFT). The rod blocks are modeled as wormlike chains and the corresponding SCFT equations are solved using a hybrid method, in which the orientation-dependent functions are discretized on a unit sphere, while the positional space-dependent functions are treated using a spectral method. Phase diagrams of the blends are constructed as a function of the homopolymer volume fraction and phase segregation strength. It is discovered that the phase behavior of the system depends on the flexibility of the homopolymers. The addition of coil-homopolymers stabilizes the smectic phases. Low-molecular weight coil-homopolymers tend to mix with the coil-blocks, whereas high-molecular weight coil-homopolymers are mostly localized at the center of the coil-domains. On the other hand, the addition of rod-homopolymers strongly affects the orientation ordering of the system, leading to transitions between monolayer smectic-C, monolayer smectic-A and bilayer smectic-A phases. PMID- 21644582 TI - Chiral biscinchona alkaloid promoted asymmetric allylic alkylation of 3 substituted benzofuran-2(3H)-ones with Morita-Baylis-Hillman carbonates. AB - A highly diastereo- and enantioselective asymmetric allylic alkylation reaction with respect to prochiral 3-substituted benzofuran-2(3H)-ones and MBH carbonate by a chiral biscinchona alkaloid catalyst was investigated. The corresponding adducts, containing a quaternary center at the C3-position of the benzofuran 2(3H)-one as well as a vicinal tertiary center, were generally obtained in high yields (up to 97%) with very good diastereo- (up to 98:2 dr) and enantioselectivities (up to 95% ee). PMID- 21644583 TI - Geometry and spectral properties of the protonated homodimer of pyridine in the liquid and solid states. A combined NMR, X-ray diffraction and inelastic neutron scattering study. AB - The structure and spectral signatures of the protonated homodimer of pyridine in its complex with a poorly coordinating anion have been studied in solution in CDF(3)/CDClF(2) down to 120 K and in a single crystal. In both phases, the hydrogen bond is asymmetric. In the solution, the proton is involved in a fast reversible transfer that determines the multiplicity of NMR signals and the sign of the primary H/D isotope effect of --0.95 ppm. The proton resonates at 21.73 ppm that is above any value reported in the past and is indicative of a very short hydrogen bond. By combining X-ray diffraction analysis with model computations, the position of the proton in the crystal has been defined as d(N H) = 1.123 A and d(H...N) = 1.532 A. The same distances have been estimated using a (15)N NMR correlation. The frequency of the protonic out-of-plane bending mode is 822 cm(-1) in agreement with Novak's correlation. PMID- 21644584 TI - Total synthesis of pinnatoxins A and G and revision of the mode of action of pinnatoxin A. AB - Pinnatoxins belong to an emerging class of potent marine toxins of the cyclic imine group. Detailed studies of their biological effects have been impeded by unavailability of the complex natural product from natural sources. This work describes the development of a robust, scalable synthetic sequence relying on a convergent strategy that delivered a sufficient amount of the toxin for detailed biological studies and its commercialization for use by other research groups and regulatory agencies. A central transformation in the synthesis is the highly diastereoselective Ireland-Claisen rearrangement of a complex alpha,alpha disubstituted allylic ester based on a unique mode for stereoselective enolization through a chirality match between the substrate and the lithium amide base. With synthetic pinnatoxin A, a detailed study has been performed that provides conclusive evidence for its mode of action as a potent inhibitor of nicotinic acetylcholine receptors selective for the human neuronal alpha7 subtype. The comprehensive electrophysiological, biochemical, and computational studies support the view that the spiroimine subunit of pinnatoxins is critical for blocking nicotinic acetylcholine receptor subtypes, as evidenced by analyzing the effect of a synthetic analogue of pinnatoxin A containing an open form of the imine ring. Our studies have paved the way for the production of certified standards to be used for mass-spectrometric determination of these toxins in marine matrices and for the development of tests to detect these toxins in contaminated shellfish. PMID- 21644585 TI - Interactions between charged surfaces with ionizable sites. AB - A key factor controlling the interactions between surfaces in aqueous solutions is the surface charge density. Surfaces typically become charged though a titration process where surface groups can become ionized based on their dissociation constant and the pH of the solution. In this work, we use a Monte Carlo method to treat this process in a system with two planar surfaces with explicitly described ionizable sites in a salt solution. We focus on a system with a surface density of ionizable sites set to 4.8 nm(-2), corresponding to silica. We find that the surface charge density changes as the surfaces come close to contact due to interactions between the ionizable groups on each surface. In addition, we observe an attraction between the surfaces above a threshold surface charge, in good agreement with previous theoretical predictions based on uniformly charged surfaces. However, close to contact we find the force is significantly different than for the uniformly charged case. PMID- 21644586 TI - Synthesis, microwave spectrum, and dipole moment of allenylisocyanide (H2C?C?CHNC), a compound of potential astrochemical interest. AB - An improved synthesis of a compound of potential astrochemical interest, allenylisocyanide (H(2)C?C?CHNC), is reported together with its microwave spectrum, which has been investigated in the 8-120 GHz spectral range to facilitate a potential identification in interstellar space. The spectra of the ground vibrational state and of five vibrationally excited states belonging to three different vibrational modes have been assigned for the parent species. A total of 658 transitions with a maximum value of J = 71 were assigned for the ground state and accurate values obtained for the rotational and quartic centrifugal distortion constants. The spectra of five heavy-atom ((13)C and (15)N) isotopologues were also assigned. The dipole moment was determined to be MU(a) = 11.93(16) * 10(-30) C m, MU(b) = 4.393(44) * 10(-30) C m, and MU(tot) = 12.71(16) * 10(-30) C m. The spectroscopic work has been augmented by theoretical calculations at the CCSD/cc-pVTZ and B3LYP/cc-pVTZ levels of theory. The theoretical calculations are generally in good agreement with the experimental results. PMID- 21644587 TI - The impact of Texas red on lipid bilayer properties. AB - We investigated the impact of fluorescent labeling on the properties of a lipid bilayer using atomistic molecular dynamics simulation. The system consisted of 24 Texas Red-1,2-dihexadecanoyl-sn-glycero-3-phophoethanolamine (TR-DHPE) in a bilayer of 488 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) lipids. We found binding between TR-DHPE and DPPC caused by electrostatic interactions. On average, TR-DHPE is bound to 1.2 DPPC molecules. Binding reduced the diffusion coefficient of TR-DHPE by 34% relative to unlabeled DPPC molecules. We estimate that binding would lead to a ~ 5 degrees C increase in the liquid to liquid ordered transition temperature of a ternary lipid system. These results emphasize the importance of considering the impact of fluorescence labeling when interpreting experimental results. PMID- 21644588 TI - Influence of deep frying on the unsaponifiable fraction of vegetable edible oils enriched with natural antioxidants. AB - The influence of deep frying, mimicked by 20 heating cycles at 180 degrees C (each cycle from ambient temperature to 180 degrees C maintained for 5 min), on the unsaponifiable fraction of vegetable edible oils represented by three characteristic families of compounds (namely, phytosterols, aliphatic alcohols, and triterpenic compounds) has been studied. The target oils were extra virgin olive oil (with intrinsic content of phenolic antioxidants), refined sunflower oil enriched with antioxidant phenolic compounds isolated from olive pomace, refined sunflower oil enriched with an autoxidation inhibitor (dimethylpolysiloxane), and refined sunflower oil without enrichment. Monitoring of the target analytes as a function of both heating cycle and the presence of natural antioxidants was also evaluated by comparison of the profiles after each heating cycle. Identification and quantitation of the target compounds were performed by gas cromatography-mass spectrometry in single ion monitoring mode. Analysis of the heated oils revealed that the addition of natural antioxidants could be an excellent strategy to decrease degradation of lipidic components of the unsaponifiable fraction with the consequent improvement of stability. PMID- 21644589 TI - Rational design and one-step formation of multifunctional gel transducer for simple fabrication of integrated electrochemical biosensors. AB - This study demonstrates a new strategy to simplify the biosensor fabrication and thus minimize the biosensor-to-biosensor deviation through rational design and one-step formation of a multifunctional gel electronic transducer integrating all elements necessitated for efficiently transducing the biorecognition events to signal readout, by using glucose dehydrogenase (GDH) based electrochemical biosensor as an example. To meet the requirements for preparing integrated biosensors and retaining electronic and ionic conductivities for electronically transducing process, ionic liquids (ILs) with enzyme cofactor (i.e., oxidized form of nicotinamide adenine dinucleotide) as the anion were synthesized and used to form a bucky gel with single-walled carbon nanotubes, in which methylene green electrocatalyst was stably encapsulated for the oxidation of nicotinamide adenine dinucleotide. With such kind of rationally designed and one-step-formed multifunctional gel as the electronic transducer, the GDH-based electrochemical biosensors were simply fabricated by polishing the electrodes onto the gel followed by enzyme immobilization. This capability greatly simplifies the biosensor fabrication, prolongs the stability of the biosensors, and, more remarkably, minimizes the biosensor-to-biosensor deviation. The relative standard deviations obtained both with one electrode for the repeated measurements of glucose and with the different electrodes prepared with the same method for the concurrent measurements of glucose with the same concentration were 3.30% (n = 7) and 4.70% (n = 6), respectively. These excellent properties of the multifunctional gel-based biosensors substantially enable them to well-satisfy the pressing need of rapid measurements, for example, environmental monitoring, food analysis, and clinical diagnoses. PMID- 21644591 TI - Effect of oxidation on surface-enhanced Raman scattering activity of silver nanoparticles: a quantitative correlation. AB - We quantitatively studied, using X-ray photoelectron spectroscopy (XPS), oxidation of substrate-immobilized silver nanoparticles (Ag NPs) in a wide range of conditions, including exposure to ambient air and controlled ozone environment under UV irradiation, and we correlated the degree of silver oxidation with surface-enhanced Raman scattering (SERS) enhancement factors (EFs). The SERS activity of pristine and oxidized Ag NPs was assessed by use of trans-1,2-bis(4 pyridyl)ethylene (BPE) and sodium thiocynate as model analytes at the excitation wavelength of 532 nm. Our study showed that the exposure of Ag NPs to parts per million (ppm) level concentrations of ozone led to the formation of Ag(2)O and orders of magnitude reduction in SERS EFs. Such an adverse effect was also notable upon exposure of Ag NPs under ambient conditions where ozone existed at parts per billion (ppb) level. The correlated XPS and SERS studies suggested that formation of just a submonolayer of Ag(2)O was sufficient to decrease markedly the SERS EF of Ag NPs. In addition, studies of changes in plasmon absorption bands pointed to the chemical enhancement as a major reason for deterioration of SERS signals when substrates were pre-exposed to ambient air, and to a combination of changes in chemical and electromagnetic enhancements in the case of substrate pre-exposure to elevated ozone concentrations. Finally, we also found UV irradiation and ozone had a synergistic effect on silver oxidation and thus a detrimental effect on SERS enhancement of Ag NPs and that such oxidation effects were analyte-dependent, as a result of inherent differences in chemical enhancements and molecular binding affinities for various analytes. PMID- 21644592 TI - Fabrication and characterization of an all-diamond tubular flow microelectrode for electroanalysis. AB - The development of the first all-diamond hydrodynamic flow device for electroanalytical applications is described. Here alternate layers of intrinsic (insulating), conducting (heavily boron doped), and intrinsic polycrystalline diamond are grown to create a sandwich structure. By laser cutting a hole through the material, it is possible to produce a tubular flow ring electrode of a characteristic length defined by the thickness of the conducting layer (for these studies ~90 MUm). The inside of the tube can be polished to 17 +/- 10 nm surface roughness using a diamond impregnanted wire resulting in a coplanar, smooth, all diamond surface. The steady-state limiting current versus volume flow rate characteristics for the one electron oxidation of FcTMA(+) are in agreement with those expected for laminar flow in a tubular electrode geometry. For dopamine detection, it is shown that the combination of the reduced fouling properties of boron doped diamond, coupled with the flow geometry design where the products of electrolysis are washed away downstream of the electrode, completely eradicates fouling during electrolysis. This paves the way for incorporation of this flow design into online electroanalytical detection systems. Finally, the all diamond tubular flow electrode system described here provides a platform for future developments including the development of ultrathin ring electrodes, multiple apertures for increased current response, and multiple, individually addressable ring electrodes incorporated into the same flow tube. PMID- 21644593 TI - Electrochemical oxidation by square-wave potential pulses in the imitation of oxidative drug metabolism. AB - Electrochemistry combined with mass spectrometry (EC-MS) is an emerging analytical technique in the imitation of oxidative drug metabolism at the early stages of new drug development. Here, we present the benefits of electrochemical oxidation by square-wave potential pulses for the oxidation of lidocaine, a test drug compound, on a platinum electrode. Lidocaine was oxidized at constant potential and by square-wave potential pulses with different cycle times, and the reaction products were analyzed by liquid chromatography-mass spectrometry [LC MS(/MS)]. Application of constant potentials of up to +5.0 V resulted in relatively low yields of N-dealkylation and 4-hydroxylation products, while oxidation by square-wave potential pulses generated up to 50 times more of the 4 hydroxylation product at cycle times between 0.2 and 12 s (estimated yield of 10%). The highest yield of the N-dealkylation product was obtained at cycle times shorter than 0.2 s. Tuning of the cycle time is thus an important parameter to modulate the selectivity of electrochemical oxidation reactions. The N-oxidation product was only obtained by electrochemical oxidation under air atmosphere due to reaction with electrogenerated hydrogen peroxide. Square-wave potential pulses may also be applicable to modulate the selectivity of electrochemical reactions with other drug compounds in order to generate oxidation products with greater selectivity and higher yield based on the optimization of cycle times and potentials. This considerably widens the scope of direct electrochemistry-based oxidation reactions for the imitation of in vivo oxidative drug metabolism. PMID- 21644594 TI - Determination of the degree of acetylation of chitin/chitosan by pyrolysis-gas chromatography in the presence of oxalic Acid. AB - A new method to determine directly and rapidly the degree of acetylation of chitin/chitosan was developed based on reactive pyrolysis-gas chromatography in the presence of an oxalic acid aqueous solution. The degree of acetylation was precisely evaluated on the basis of peak intensities of the characteristic products such as acetonitrile, acetic acid, and acetamide originating from the N acetyl group of N-acetyl-d-glucosamine units of chitin/chitosan. The observed values were in good agreement with those obtained by (1)H NMR and the other methods. Moreover, the proposed technique was applicable to any kinds of chitin/chitosan samples over the whole range of acetylation including insoluble chitin/chitosan and perfectly acetylated artificial chitin having higher crystallinity to which (1)H NMR had been inapplicable. PMID- 21644595 TI - Strategies for the analysis of polar solvents in liquid matrixes. AB - Various approaches to the analysis of polar compounds in different matrixes by solid-phase microextraction (SPME) were studied. The analysis of polar analytes in nonpolar matrixes was performed with custom-made SPME fibers coated with Nafion perfluorinated resin. The sensitivity of this fiber in this type of analysis was better by 1 order of magnitude on average as compared to those of any of the commercially available fibers. The fiber was the most sensitive for the most polar of the compounds studied, i.e., methanol. Determination of methanol, ethanol, and 2-propanol in unleaded gasoline was illustrated. Except for methanol, the fiber did not perform very well in the analysis of alcohols in water. The fiber was capable of extracting water from benzene. SPME analysis of polar compounds in water was studied using aqueous solutions of acetone, methyl ethyl ketone, methyl isobutyl ketone (MIBK), 2-propanol, 2-methyl-2-propanol, and tetrahydrofuran. Fibers coated with poly(dimethylsiloxane)/divinylbenzene yielded the highest sensitivity in this type of analysis. Low- or sub-ppb detection limits were obtained for all the analytes with FID detection when the samples were saturated with NaCl. Since fibers of this type extract analytes by adsorption rather than absorption, nonlinear responses were observed when all the analytes were allowed to equilibrate because of the limited number of adsorption sites on the surface of the coating and displacement of compounds with low distribution ratios by compounds with high distribution ratios (mainly MIBK). Two approaches allowed a significant improvement in linearity: extraction of a vigorously stirred sample for a short time, or extraction under static conditions for a time much shorter than that required for equilibration of all the analytes. In both cases the amount of MIBK extracted was significantly reduced, while the remaining analytes were affected to a much lesser degree. The sensitivity of acetone determination was greatly improved by in-solution derivatization with o (2,3,4,5,6-pentafluorobenzyl)hydroxylamine hydrochloride and extraction of the oxime formed. PMID- 21644596 TI - Quantitative retention-structure and retention-activity relationship studies of local anesthetics by micellar liquid chromatography. AB - The retention of compounds in micellar liquid chromatography (MLC) is governed by hydrophobic and electrostatic forces. For ionic compounds, both interactions should be considered. The present report offers a novel retention model that includes the hydrophobicity of compounds and the molar fraction of the charged form of compounds and compares it with other previously reported models. High correlations between the logarithm of capacity factors and these structural parameters were obtained for local anesthetics with different degrees of ionization using a nonionic surfactant solution as mobile phase. Modeling the retention of compounds as a function of physicochemical parameters and experimental variables is established by means of multiple linear regression. In addition, a predictive model for estimating the hydrophobicity of local anesthetics is proposed. Finally, quantitative and qualitative retention-activity relationships in MLC are also investigated for these compounds. An excellent correlation between the capacity factors in MLC and the anesthetic potency of local anesthetics was obtained. PMID- 21644597 TI - Theoretical Justification of Wavelength Selection in PLS Calibration: Development of a New Algorithm. AB - The mathematical basis of improved calibration through selection of informative variables for partial least-squares calibration has been identified. A theoretical investigation of calibration slopes indicates that including uninformative wavelengths negatively affect calibrations by producing both large relative bias toward zero and small additive bias away from the origin. These theoretical results are found regardless of the noise distribution in the data. Studies are performed to confirm this result using a previously used selection method compared to a new method, which is designed to perform more appropriately when dealing with data having large outlying points by including estimates of spectral residuals. Three different data sets are tested with varying noise distributions. In the first data set, Gaussian and log-normal noise was added to simulated data which included a single peak. Second, near-infrared spectra of glucose in cell culture media taken with an FT-IR spectrometer were analyzed. Finally, dispersive Raman Stokes spectra of glucose dissolved in water were assessed. In every case considered here, improved prediction is produced through selection, but data with different noise characteristics showed varying degrees of improvement depending on the selection method used. The practical results showed that, indeed, including residuals into ranking criteria improves selection for data with noise distributions resulting in large outliers. It was concluded that careful design of a selection algorithm should include consideration of spectral noise distributions in the input data to increase the likelihood of successful and appropriate selection. PMID- 21644598 TI - Tailoring of sol-gel films for optical sensing of oxygen in gas and aqueous phase. AB - Sol-gel-based optical sensors for both gas-phase and dissolved oxygen have been developed. Both sensors operate on the principle of fluorescence quenching of a ruthenium complex which has been entrapped in a porous sol-gel silica film. A comprehensive investigation was carried out in order to establish optimal film processing parameters for the two sensing environments. Both tetraethoxysilane and organically modified sol-gel precursors such as methyltriethoxysilane and ethyltriethoxysilane were used. Film hydrophobicity increases as a function of modified precursor content, and this was correlated with enhanced dissolved oxygen (DO) sensor performance. Extending the aliphatic group of the modified precursor further improved DO sensitivity. The influence of water/precursor molar ratio, R, on the sol-gel film microstructure was investigated. R value tailoring of the microstructure and film surface hydrophobicity tailoring were correlated with oxygen diffusion behavior in the films via the Stern-Volmer constants for both gas phase and DO sensing. Excellent performance characteristics were measured for both gas-phase and DO oxygen sensors. The long-term quenching stability of DO sensing films was established over a period of 6 months. PMID- 21644599 TI - Effects of sample dimension and dye distribution characteristics in absorption microspectroscopy. AB - Models and the relevant equations giving optical absorbance of an analyte (i.e., dye) confined in a small volume with different shapes and sizes (sample dimension, D) are given, and factors governing absorbance are discussed. Dye absorbance is shown to be dependent on the three-dimensional sample structure: films (1-D), tubes (2-D), and spheres (3-D). The magnitude of the dimension effect on absorbance is determined by the size of both the sample and the probe beam. Furthermore, dye distribution characteristics in 2-D and 3-D systems also affect absorbance; a dye distributed exclusively to the surface layer of a 2-D or 3-D sample gave an absorbance 0.50- or 0.33-fold of that homogeneously distributed, respectively. If a dye is distributed in the inner volume of the sample alone, absorbance increases over that for homogeneous distribution. Effects of sample dimension, dye distribution characteristics, and the size of the probe beam in absorption microspectroscopy are discussed on the basis of the proposed models, and an experimental check of the models is presented. PMID- 21644600 TI - Increasing the precision and accuracy of top-loading balances: application of experimental design. AB - The traditional method of estimating the weight of multiple objects is to obtain the weight of each object individually. We demonstrate that the precision and accuracy of these estimates can be improved by using a weighing scheme in which multiple objects are simultaneously on the balance. The resulting system of linear equations is solved to yield the weight estimates for the objects. Precision and accuracy improvements can be made by using a weighing scheme without requiring any more weighings than the number of objects when a total of at least six objects are to be weighed. It is also necessary that multiple objects can be weighed with about the same precision as that obtained with a single object, and the scale bias remains relatively constant over the set of weighings. Simulated and empirical examples are given for a system of eight objects in which up to five objects can be weighed simultaneously. A modified Plackett-Burman weighing scheme yields a 25% improvement in precision over the traditional method and implicitly removes the scale bias from seven of the eight objects. Applications of this novel use of experimental design techniques are shown to have potential commercial importance for quality control methods that rely on the mass change rate of an object. PMID- 21644601 TI - Measurement of cosmogenic (32)p and (33)p activities in rainwater and seawater. AB - We have developed a new method for the collection, purification, and measurement of natural levels of (32)P and (33)P in rain, marine particulates, and dissolved constituents of seawater. (32)P and (33)P activities were measured using a recently developed ultra-low-level liquid scintillation counter. Measurement by liquid scintillation counting allows, for the first time, simultaneous measurement of both (32)P and (33)P. Furthermore, (33)P activities are measured with high efficiency (>50%), regardless of the amount of stable phosphorus in the sample. Liquid scintillation also produces energy specific beta spectra which has enabled us to identify previously unrecognized beta-emitting contaminants in natural samples. In order to remove these contaminants, new methods of purification have been developed which utilize a series of precipitations and anion and cation exchange columns. Rainwater and dissolved seawater samples were extracted from large volumes of rain- and seawater, 5-20 and >5000 L, respectively, using iron-impregnated polypropylene filters. On these filters, it was possible to load between 25 and 30% Fe(OH)(3) by weight, over twice that loaded on previously utilized materials. Using our collection, purification, and liquid scintillation counting techniques, it was possible to obtain specific (32)P and (33)P activities with less than 10% error (2sigma) in rainwater and 20% error (2sigma) in seawater. PMID- 21644602 TI - Multivariate optical computation for predictive spectroscopy. AB - A novel optical approach to predicting chemical and physical properties based on principal component analysis (PCA) is proposed and evaluated using a data set from earlier work. In our approach, a regression vector produced by PCA is designed into the structure of a set of paired optical filters. Light passing through the paired filters produces an analog detector signal that is directly proportional to the chemical/physical property for which the regression vector was designed. This simple optical computational method for predictive spectroscopy is evaluated in several ways, using the example data for numeric simulation. First, we evaluate the sensitivity of the method to various types of spectroscopic errors commonly encountered and find the method to have the same susceptibilities toward error as standard methods. Second, we use propagation of errors to determine the effects of detector noise on the predictive power of the method, finding the optical computation approach to have a large multiplex advantage over conventional methods. Third, we use two different design approaches to the construction of the paired filter set for the example measurement to evaluate manufacturability, finding that adequate methods exist to design appropriate optical devices. Fourth, we numerically simulate the predictive errors introduced by design errors in the paired filters, finding that predictive errors are not increased over conventional methods. Fifth, we consider how the performance of the method is affected by light intensities that are not linearly related to chemical composition (as in transmission spectroscopy) and find that the method is only marginally affected. In summary, we conclude that many types of predictive measurements based on use of regression (or other) vectors and linear mathematics can be performed more rapidly, more effectly, and at considerably lower cost by the proposed optical computation method than by traditional dispersive or interferometric instrumentation. Although our simulations have used Raman experimental data, the method is equally applicable to Near-IR, UV-vis, IR, fluorescence, and other spectroscopies. PMID- 21644603 TI - Comparison of Potential-Time Waveforms for the Detection of Biogenic Amines in Complex Mixtures following Their Separation by Liquid Chromatography. AB - Pulsed amperometric detection (PAD), integrated voltammetric detection (IVD), and integrated square-wave detection (ISWD) at gold electrodes are compared for the flow injection detection of 1,3-diaminopropane in a liquid chromatograph (LC). These detection methods are especially significant for alkylamines and amino acids because (i) the majority of these compounds do not naturally possess chromophoric or fluorophoric functional groups and (ii) their amperometric detection at constant potential at Au electrodes fails because the electrode activity is severely attenuated by the formation of an inert surface oxide (AuO). The anodic response mechanisms for detection of amines require concomitant formation of AuO; therefore, a large background signal is observed with conventional PAD. In comparison, the background is much smaller for IVD and ISWD because the anodic charge for oxide formation (positive scan/step) is compensated by the cathodic charge for oxide reduction (negative scan/step). The limits of detection (S/N = 3) for 1,3-diaminopropane by LC-PAD, LC-IVD and LC-ISWD are about 3 * 10(2) pg (4 pmol), 5 * 10(1) pg (0.7 pmol), and 3 * 10(1) pg (0.5 pmol), respectively, for 25-MUL injections. Results are also shown to demonstrate ISWD for detection of nine biogenic amines following their chromatographic separation. PMID- 21644604 TI - Column design for electrochemically modulated liquid chromatography. AB - A new column design for electrochemically modulated liquid chromatography (EMLC) is presented. The principal attribute of the new design is the enhancement of the control over the potential applied to the stationary phase by the reduction of the background current and solution resistance. The enhancement is demonstrated through a series of comparisons of the electrochemical performance of the new and the earlier column designs. This enhancement, as shown using mixtures of aromatic sulfonates, translates to an improvement in the capability of EMLC as a separation technique. PMID- 21644605 TI - Direct excitation energy transfer as a technique for in situ measurements of ion exchange processes in single polymer particles. AB - Ion-exchange processes of a cationic dye (Malachite Green, MG) are studied for individual polymer particles (diameter of 20 or 13 MUm) by laser trapping microspectroscopy. When a cation-exchange resin, preadsorbed homogeneously with Rhodamine B (RhB), is soaked in an aqueous MG solution, MG is adsorbed in the surface layer of the particle in the initial stage and then diffuses into the inner volume with time. In the MG-diffused layer, RhB fluorescence is quenched by excitation energy transfer from the excited state of RhB to the ground-state MG, as revealed by fluorescence microspectroscopy of individual resin particles in the dye solution. On the basis of the time dependence of the quenching efficiency, the thickness of the MG-diffused layer at a given soaking time is estimated, and the diffusion coefficient of the dye in the particle is determined to be 8 * 10(-)(11) cm(2) s(-)(1). This method is shown to be very useful for determining the diffusion coefficient of a nonfluorescent ion in individual ion exchange resin microparticles. PMID- 21644606 TI - Microspectroscopic analyses of dye distribution characteristics in single microcapsules. AB - The distribution characteristics of a dye in single melamine resin wall microcapsules containing a dye/toluene solution are studied by a laser trapping absorption microspectroscopy technique. In the case of Disperse Orange 13 as a dye, the molar absorptivity determined from the slope of a dye absorbance-capsule diameter (d) plot agrees well with that observed in a homogeneous toluene solution, indicating that the dye is solubilized homogeneously in the inner toluene solution of the capsule. For tetraphenylporphyrins (MTPP: Zn(II)TPP, Co(II)TPP, or H(2)TPP), however, the d dependence of absorbance shows a sigmoidal curve. Since absorbance of a ZnTPP/toluene droplet before polymerization of the melamine resin increases linearly with an increase in the droplet diameter, the sigmoidal d dependence of the absorbance is concluded to originate from distribution of the dye into the resin wall. Detailed analyses of the data indicate that the partitioning ratio of the dye between the toluene and resin phases is dependent on the capsule diameter. Size-dependent distribution of MTPP in single microcapsules is discussed on the basis of a proposed model and the d dependence of the dye absorbance, and it is concluded that the distribution of MTPP to the melamine resin wall is facilitated for larger microcapsules. PMID- 21644607 TI - Tandem mass spectrometry of uranium and uranium oxides in airborne particulates. AB - A method for detection of uranium in airborne microparticles in real time has been developed. Positive identification of uranium is achieved by isolating UO(2+) ions and following their reaction with residual oxygen molecules to yield UO(2)(+). PMID- 21644608 TI - On-line concentration of neutral analytes for micellar electrokinetic chromatography. 3. Stacking with reverse migrating micelles. AB - On-line concentration of neutral analytes by sample stacking in reversed migration micellar electrokinetic chromatography is presented. Micellar separation solutions of sodium dodecyl sulfate are prepared with acidic buffers to reverse the direction of the migration velocity of neutral analytes owing to a reduced electroosmotic flow. Samples are prepared in nonmicellar matrixes of low conductivity (i.e., water, diluted buffer, or dilute organic/aqueous solvent) to achieve field enhancement in the sample zone. Without polarity switching inherent in large-volume sample stacking, narrowing of analyte bands, removal of sample matrix, and separation of focused analyte bands are achieved. A model is proposed to describe the stacking technique and is supported by experimental results. In addition, equations are derived to describe band broadening associated with the technique. Detector response improvements reaching a 100-fold are confirmed experimentally. Concentration detection limits on the order of low-ppb levels (S/N = 3) are realized with model steroidal compounds. PMID- 21644609 TI - Prediction of electrophoretic mobilities. 1. Monoamines. AB - The mobility of an ion is of fundamental importance in capillary electrophoresis. The size, shape, and other physicochemical parameters of monoamines are determined using molecular modeling. These parameters are used to generate regression expressions to predict absolute (infinite dilution) mobilities. Molecular volume or mass is the strongest determinant of electrophoretic mobility. However, molecular volumes calculated via molecular modeling varied systematically depending on the software used, and so molecular mass is the favored descriptor. Neither the classical spherical (Huckel) nor ellipsoidal (Perrin) models were reasonable predictors of mobility. In accord with empirical expressions, such as the Wilke-Chang equation for diffusion, the absolute mobilities correlate with mass (or volume) to a much greater power than predicted by Stokes's law. Incorporation of the effects of hydration using the McGowan waters of hydration increments further improved the predictions. The best equation for predicting absolute mobilities of monoamines is MU(0) = [(5.55 +/- 0.73) * 10(-)(3)]/[W((0.579)(+/-)(0.026)) + (0.171 +/- 0.054)H] where W is the molecular weight and H is the mean waters of hydration calculated using the McGowan increments. The uncertainties are the standard deviations of the parameters. This equation yielded an average prediction error of 4.1% for the data set used to generate the expression (literature absolute mobilities for 34 monoamines possessing no other functional groups), 7.2% for an independent data set from the literature (absolute mobilities for seven monoamines possessing other functional groups), and 3.3% for an experimentally determined data set (13 monoamines determined using capillary electrophoresis). PMID- 21644610 TI - Evaluation of a sheath flow cuvette for postcolumn fluorescence derivatization of DNA fragments separated by capillary electrophoresis. AB - The investigation and evaluation of the sheath flow cell as a reaction chamber to postcolumn fluorescently derivatize DNA fragments separated by capillary electrophoresis is described herein. Use of the sheath flow cell arrangement facilitates the mixing of the intercalating dye, ethidium bromide (EB), and the effluent from the separation capillary by diffusion without a high degree of band dispersion. Theoretical plate counts of >1 * 10(6) are reported with the postcolumn derivatization technique, and resolution of all of the fragments in a phix-174-HaeIII digest is achieved. Optimization of experimental parameters such as flow rate, position of the detection zone, and EB concentration is examined. A limit of detection in the low nanograms-per-milliliter range with a linear dynamic range over 3 orders of magnitude is reported for a sample of phix-174 HaeIII digest. Evaluation of postcolumn derivatization for the investigation of DNA-protein interactions is demonstrated. The integrity of a DNA-trp-repressor protein interaction is maintained with the postcolumn approach but is compromised when EB is added to the running buffer. PMID- 21644611 TI - Evaluation of Column Cleanup for Chlorobenzenes, Polychlorinated Biphenyls, Polychlorinated Dibenzo-p-dioxins, and Polychlorinated Dibenzofurans in MM5 Flue Gas Analysis. AB - The recoveries of chlorobenzenes (CBs), polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs) from the acid-base-on-silica column (ABS), the silver nitrate-on-silica column (SNS), and the alumina column (ALM) were investigated using eluents with various concentrations of dichloromethane in hexane. PCBs recoveries from the ABS and ALM columns are generally increased with increasing number of chloro substituents. However, PCBs recoveries are also complicated by the inductive and resonance contributions due to the different conformation and electron density of the congeners. The recoveries of PCBs with lower number of chloro substituents decreased with increasing silver nitrate content coated on silica. With 2% dichloromethane in hexane (as normally used in MM5 analysis), the recoveries of all CBs and trichloro- to decachlorobiphenyls from ABS, 10% SNS, and ALM columns in series are >=80%. However, the 2,4-dichlorobiphenyl is only 16% recovered, and monochlorobiphenyls are not recovered at all. Under these conditions, the recoveries of PCDDs/PCDFs from ABS and 10% SNS columns in series are >=80%. With 60% dichloromethane in hexane, PCDDs/PCDFs recoveries from ALM column are >=80%. The implication of the selective adsorption of PCBs on the fractionation of CBs/PCBs and PCDDs/PCDFs from these MM5 cleanup columns is discussed. PMID- 21644612 TI - Comparisons of polymer/gas partition coefficients calculated from responses of thickness shear mode and surface acoustic wave vapor sensors. AB - Apparent partition coefficients, K, for the sorption of toluene by four different polymer thin films on thickness shear mode (TSM) and surface acoustic wave (SAW) devices are compared. The polymers examined were poly(isobutylene) (PIB), poly(epichlorohydrin) (PECH), poly(butadiene) (PBD), and poly(dimethylsiloxane) (PDMS). Independent data on partition coefficients for toluene in these polymers were compiled for comparison, and TSM sensor measurements were made using both oscillator and impedance analysis methods. K values from SAW sensor measurements were about twice those calculated from TSM sensor measurements when the polymers were PIB and PECH, and they were also at least twice the values of the independent partition coefficient data, which is interpreted as indicating that the SAW sensor responds to polymer modulus changes as well as to mass changes. K values from SAW and TSM measurements were in agreement with each other and with independent data when the polymer was PBD. Similarly, K values from the PDMS coated SAW sensor were not much larger than values from independent measurements. These results indicate that modulus effects were not contributing to the SAW sensor responses in the cases of PBD and PDMS. However, K values from the PDMS coated TSM device were larger than the values from the SAW device or independent measurements, and the impedance analyzer results indicated that this sensor using our sample of PDMS at the applied thickness did not behave as a simple mass sensor. Differences in behavior among the test polymers on SAW devices are interpreted in terms of their differing viscoelastic properties. PMID- 21644613 TI - The Use of Gas-Phase UV Spectra in the 168-330-nm Wavelength Region for Analytical Purposes. 1. Qualitative Measurements. AB - A novel instrument, which combines gas chromatographic separation with ultraviolet detection, has been employed in the qualitative study of molecular UV absorption spectra in the gas phase. The wavelength range available with the current instrument made studies in the region of 168-330 nm possible. The absorption spectra between 168 and 330 nm for ~1000 organic compounds and a number of inorganic gases were registered. For all the spectra recorded, 69% showed absorption maxima shorter than 190 nm, and as much as 87% shorter than 200 nm. This indicates the importance of the short UV wavelength region for analytical purposes. The results showed that each compound studied gave rise to unique absorption spectra. The influence of temperature on the shape of the spectra was studied in the range of 15-205 degrees C. A slight broadening effect on spectral absorption bands (0.3 nm) and the vibrational structure (maximally 1.4 nm) with increased temperature was observed. However, this effect was shown to have no influence on the analytical results. In the vast majority of cases, where structural isomers were investigated, there were clear differences between the recorded spectra. Compounds with the same functional group showed pronounced similarities. The influence of conjugation and various substituents could be demonstrated. A commercial computer program was used for a computer search of unknowns against the recorded reference spectra. This showed that the gas-phase UV spectra are very well defined and that a search always resulted in a very close agreement, even if concentrations and chromatographic conditions were different between the recording of reference and unknown. When a search was performed on a compound not present in the library, the result pointed out the correct class of compounds. PMID- 21644614 TI - Hardware modifications to an isotope ratio mass spectrometer continuous-flow interface yielding improved signal, resolution, and maintenance. AB - A home-built combustion interface was constructed to improve signal, resolution, and maintenance of a continuous-flow gas isotope ratio system. Chromatographic peak shapes were preserved by minimizing changes in tubing diameter and dead volumes. A single piece of fused silica capillary was used to connect the gas chromatograph (GC) to the isotope ratio mass spectrometer (IRMS), thus eliminating extraneous combustion furnace and water trap fittings. Analysis of a standard mixture of hydrocarbons yielded a 2-fold increase in signal over a slightly modified conventional system. Column efficiency, expressed as trennzahl (TZ), improved significantly (Student's t-test 95% CI) by an average factor of 1.4 for replicates analyzed under similar conditions. The design is robust, requires less maintenance, and reduces leaks because the number of connections is minimized. Benefits of this system are transferable to virtually all commercially available continuous-flow systems. PMID- 21644615 TI - Electrochemical thermospray mass spectrometry instrumentation for coupling electrochemistry to mass spectrometry. AB - Reaction products in electrochemical processes can be identified by coupling an electrochemical thin-layer flow cell to a thermospray mass spectrometer. The performance of this analytical method, electrochemical thermospray mass spectrometry, is demonstrated. This includes the characterization of the improved electrochemical thin-layer flow cell. This cell offers the possibility to combine cyclic voltammograms with mass spectrometry. This goal was achieved, too, by the construction of a new thermospray ion source and a special vacuum recipient. PMID- 21644616 TI - Determination of Block Length Distributions of Poly(oxypropylene) and Poly(oxyethylene) Block Copolymers by MALDI-FTICR Mass Spectrometry. AB - Matrix-assisted laser desorption/ionization (MALDI) was performed on an external ion source Fourier transform ion cyclotron resonance mass spectrometer (FTICR-MS) to analyze the block length distributions of triblock polymers of poly(oxypropylene) and poly(oxyethylene). The first series of results presented demonstrate that the apparent molecular weight distributions are distorted. This distortion is induced by the flight-time-induced mass discrimination inherent in the experimental technique, the variation of isotopic patterns over the measured mass range, and the overlap of peaks in the spectrum. Subsequently, a method for the treatment of molecular weight distributions measured by MALDI on an external ion source FTICR-MS is developed to yield the actual molecular weight distribution and, from that, the individual block length distributions. For the first time, detailed and accurate molecular weight data were obtained on a complex sample using this methodology, which independently validates the data provided by the manufacturer. The experimentally verified random coupling hypothesis proves the validity of the methodology. PMID- 21644617 TI - Bioconcentration factors for volatile organic compounds in vegetation. AB - Samples of air and leaves were taken at the University of Nevada [Formula: see text] Las Vegas campus and analyzed for volatile organic compounds using vacuum distillation coupled with gas chromatography/mass spectrometry. The data were used to estimate the bioconcentration of volatile organic compounds (VOCs) and to characterize the equilibration of VOCs between the leaves and air. The bioconcentration of volatiles in the leaves of some species can be predicted using the partition coefficients between air and octanol (K(oa)) and only considering VOC absorption in the lipid fraction of leaves. For these leaves, the bioconcentration factors agreed with existing models. Leaves of some species displayed a bioconcentration of volatiles that greatly exceeded theory. These hyperbioconcentration leaves also contain appreciable concentrations of monoterpenes, suggesting that a terpenoid compartment should be considered for the bioconcentration of organic compounds in leaves. Adding an additional "terpenoid" compartment should improve the characterization of volatile organic compounds in the environment. The uptake of VOCs from air by leaves is rapid, and the equilibration rates are seen to be quicker for compounds that have higher vapor pressures. The release of VOCs from the leaves of plants is slower for hyperbioconcentration leaves. PMID- 21644618 TI - Multistage accurate mass spectrometry: a "basket in a basket" approach for structure elucidation and its application to a compound from combinatorial synthesis. AB - A "basket in a basket" method based on a multistage accurate mass spectrometric (MAMS) technique was developed and demonstrated by obtaining a unique elemental composition of a compound (with a molecular weight of 517) from combinatorial synthesis. The accurate masses for the parent and the fragment ions were obtained with up to five stages of MAMS using Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS). This approach requires only input of elements used in the synthetic processes and some constraints about unusual light elements, such as fluorine, while the compositions of the parent ions and their fragments are obtained for structure elucidation. Conversely, accuracy of better than 0.02 ppm (assuming elements C, H, N, O, S, and F are involved) would be required in order to define a unique composition for the same mass using a direct accurate mass measurement because the number of possible elemental compositions increases sharply as the mass increases. Similarly, due to the uncertainty in determining elemental compositions of fragments and complexity of possible internal fragmentation, tandem mass spectrometry may not provide enough information for structure elucidation of unknown compounds, especially of the organic molecules in the mass range of 300-1000 Da, typically encountered in combinatorial lead generation. The application of MAMS to combinatorial drug discovery is particularly advantageous since the built-in chemical information from the synthesis can be used as constraints. The implementation of a nanoelectrospray ionization technique makes this approach practical for characterization of small quantities of compounds typically available from lead generation processes. PMID- 21644619 TI - Solvation energy and gas-phase stability influences on alkali metal cluster ion formation in electrospray ionization mass spectrometry. AB - The ability to observe abundant gas-phase metal cluster ions in electrospray ionization mass spectrometry (ESI-MS) is highly dependent on experimental conditions. Alkali halides (MX) and other alkali metal salts were used to investigate the formation of cluster ions in ESI-MS. All compounds were found to give cluster ions of the form (M(n)(+1)X(n))(+) and (M(n)X(n+1))(-), with only two alkali salts yielding doubly charged cluster ions. In homologous alkali halide series, the relative abundances of cluster ions increased with increasing size of either the cation (positive ion mode) or the anion (negative ion mode). Calculations using an electrostatic model show that the gas-phase stability of cluster ions is greater for smaller cations or anions when a fixed counterion is employed. This stability calculation goes in a direction just opposite to the trend in cluster ion abundances observed in ESI-MS. Studies of equimolar mixtures consisting of two alkali halides reveal two distinct trends. When the equimolar mixture was composed of differing ions that participate in the droplet charge excess with the same counterion, the less solvated ions were found to form more abundant cluster ions. When the ions participating in the charge excess were fixed, the preferred counterion in observed clusters was the one that is more solvated in solution and forms more stable clusters in the gas phase. These observations can be rationalized by an extended form of the charged residue model where the weakly solvated ions that are part of the charge excess are preferentially enriched in offspring droplets during uneven fission. By contrast, transfer of a particular counterion located in the bulk of the droplets to the offspring droplets is not disfavored when this counterion is strongly solvated. PMID- 21644620 TI - Amino Acid analysis of peptides and proteins on the femtomole scale by gas chromatography/mass spectrometry. AB - Amino acid analysis (AAA) is a useful aid in protein chemistry, but its routine application is limited by a modest limit of detection. Typically, 10 pmol of material is required, but even at this level the reproducibility can be poor. We have employed isotope dilution gas chromatography electron capture negative ionization mass spectrometry (GC/ECNI/MS) to provide accurate and reliable data on less than 100 fmol of material. Precision and accuracy are good, and all 20 non-hydrolyzed amino acids can be determined in this manner. The protein is hydrolyzed (HCl), and then a cocktail, composed of all 20 amino acids as stable isotope-labeled forms (i.e., (13)C and (2)H), is added. The mixture of protein derived and stable isotope-labeled amino acids is then converted to volatile electron-capturing derivatives with a multistep approach employing heptafluorobutyric anhydride (HFBA), pentafluorobenzyl bromide (PFBBr), and N,O bis(trimethylsilyl)trifluoroacetamide (BSTFA). These derivatives are then injected directly into the GC/MS system. Groups of selected ions, characteristic of each derivatized amino acid, are thereafter monitored at appropriate time intervals. The ratios of the ion current for the selected ions for the native amino acid and its labeled form are determined and converted to absolute amounts of the native amino acids in the protein hydrolyzate by reference to standard samples prepared at the time of the analysis. PMID- 21644621 TI - A water-soluble tetraethylsulfonate derivative of 2-methylresorcinarene as an additive for capillary electrophoresis. AB - A water-soluble tetraethylsulfonate derivative of 2-methylresorcinarene (TESMR), an aromatic-based, bowl-shaped macrocycle, was used as an additive in capillary electrophoresis. Several phenol derivatives are used as analytes to demonstrate the effect of this highly charged additive. TESMR is observed to interact differently with a mixture of positional isomers and other types of phenol derivatives. A comparative study of separations with two charged additives, TESMR and sulfobutyl ether-beta-cyclodextrin (SBE-beta-CD), provided insight into the selectivity exhibited by these additives. The influence of buffer pH, ionic strength, and organic modifier content on separation and peak shape is investigated. Peak asymmetry caused by the use of highly charged additives at lower pH is minimized by the addition of small amounts of a polar aprotic organic solvent to the run buffer. The effects of mixing a charged additive with a neutral additive are also discussed. PMID- 21644622 TI - Separation of chiral biodegradation intermediates of linear alkylbenzenesulfonates by capillary electrophoresis. AB - High-performance capillary electrophoresis (HPCE) with alpha-cyclodextrin as the chiral selector was applied to separate the enantiomers of p-sulfophenyl-2 butyrate (SP2B) and p-sulfophenyl-3-butyrate (SP3B), which occur as biodegradation intermediates of linear alkylbenzenesulfonates (LAS), the widely used anionic surfactants. With this analytical method, we studied the transformation of both SP3B enantiomers in a laboratory batch incubation with activated sewage sludge of a municipal wastewater treatment plant. (S)-(+)-SP3B and (R)-(-)-SP3B could be detected in mechanically treated sewage effluent. After enrichment on graphitized carbon black (Carbopack B), the extracts were analyzed by HPLC with UV diode array and fluorescence detection as well as by HPCE with UV diode array detection. Quantification of SP3B in a 24-h composite sample of primary sewage effluent yielded 34 MUg/L (limit of detection, 0.1 MUg/L) of the racemic mixture determined by HPLC and 18 MUg/L of each enantiomer measured by HPCE (limit of detection, 1 MUg/L). PMID- 21644623 TI - Counterionic Detection by ICP-AES for Determination of Inorganic Anions in Water Elution Ion Chromatography Using a Zwitterionic Stationary Phase. AB - The analytical methods for the determination of inorganic anions by water elution ion chromatography using a zwitterionic column (C14SB-coated column) have been explored, where pure water was used as the mobile phase. In the present ion chromatography, the complicated peaks derived from various "ion pairs" appeared on the chromatogram when the sample solution contained several kinds of cations and anions. The chromatograms with the complicated peaks were simplified by using a preconditioning cation-exchange column. In the preconditioning column, various kinds of countercations of the analyte anions were converted to a particular kind of common cation, and thus all the analyte anions were separated as the common cation form. The more effective separation was achieved by converting to a divalent cation (Mg(2+)) form than a monovalent cation (Na(+)) form, because the anions paired with the divalent cation provided longer elution times than those paired with the monovalent cation. All the analyte anions separated as the common cation form were simply determined from their calibration curves drawn from conductivity detection. In addition, the counterionic determination of the anions was also attempted by using ICP-AES. The analyte anions were determined by measuring their countercations with ICP-AES, where the calibration curve of the countercation measured by ICP-AES could be used. PMID- 21644624 TI - Molecularly imprinted chiral stationary phase prepared with racemic template. AB - The first example of molecularly imprinted chiral stationary phase prepared using a racemic template is shown. N-(3,5-Dinitrobenzoyl)-alpha-methylbenzylamine (DNB) was chirally discriminated on the molecularly imprinted stationary phase prepared using racemic DNB as the template. A chiral monomer, (S)-(-)-N-methacryloyl-1 naphthylethylamine, was utilized as the functional monomer toward the racemic template, and its chiral recognition ability was, interestingly, found to be enhanced through racemic molecular imprinting. A thermodynamic discussion briefly suggests that the observed chiral recognition ability of the racemic imprinting was proper value. PMID- 21644625 TI - An automated system for near-real-time monitoring of trace atmospheric halocarbons. AB - A new gas chromatographic method developed to quantitatively determine important atmospheric halocarbons is described. Target compounds include replacement CFCs, chlorinated solvents, and biosynthesized (naturally produced) organohalogens, all trace gases in the atmosphere at concentrations ranging from 0.1 to 600 pptv (where pptv = 1 part in 10(-)(12) by volume). A combination of ultralow concentrations and relatively small electron attachment cross sections renders these compounds very difficult to routinely measure in the background air typical of remote atmospheric monitoring stations. Detection is achieved by preconcentration of a 200-mL air sample using an adsorbent-filled microtrap and enhancement of electron capture detector response by oxygen doping one of two detectors connected in series. Oxygen doping specifically targets halocarbons with relatively poor electron attachment rate coefficients. The work described here details construction of a novel analytical system, laboratory trials, and optimization followed by an extended field campaign at a remote atmospheric monitoring station, Mace Head, Ireland. A calibration standard or ambient air sample was acquired every hour using a cyclic, automated procedure without employing cryogenic preconcentration or refocusing. Overall precision of the analytical method for the target compounds is between 0.3 and 1.5%. PMID- 21644626 TI - Understanding molecular association and isomers recognition in isomer cyclodextrin multiple complex formation by improved liquid chromatographic studies. AB - Equations are derived that allow the determination of guest-cyclodextrin primary (K(11)), secondary (K(12)), and higher order (K(1)(n)) binding constants as well as the degree of complexation n (the percent of complexed guest), by monitoring the observed capacity factor k'(obs) for guests that can be structural, geometric, or optical isomers. The retention behavior (elution order) in a mixture of isomers is dependent on the cyclodextrin concentration in the mobile phase as well as on the stoichiometry and the binding constant of the guest cyclodextrin (G-CD) complex. The simplification of higher order complexes (G CD(n)) to that with 1:1 stoichiometry can lead to erroneous results; therefore, it is important for the stoichiometry to be determined accurately, prior to any binding constant calculations, following the continuous variation method. The proposed models are solved iteratively using nonlinear least-squares analysis and following specific algorithms. PMID- 21644627 TI - Sequential injection separation system with stopped-flow radiometric detection for automated analysis of (99)tc in nuclear waste. AB - An automated procedure for the determination of (99)Tc in aged nuclear waste has been developed. Using advanced sequential injection (SI) analysis instrumentation, (99)Tc(VII) is separated from radioactive and stable interferences using a TEVA resin column that selectively retains pertechnetate ion from dilute nitric acid solutions. The separated (99)Tc is eluted with 6 M nitric acid and quantified on-line with a flow-through liquid scintillation detector. A stopped-flow technique has been optimized that improves the analysis precision and detection limit compared to continuous-flow detection, reduces consumption of liquid scintillation cocktail, and increases sample throughput by separating the next sample while the present sample is being counted. The detection limit is 30 pCi, or 2 ng, of (99)Tc, using a 15-min stopped-flow period. The analysis time is 40 min for the first sample and is reduced to 20 min for each subsequent sample. Processed nuclear waste samples from the Hanford site were successfully analyzed by this new method. PMID- 21644628 TI - Cylindrically etched carbon-fiber microelectrodes for low-noise amperometric recording of cellular secretion. AB - The most important sources of noise with disk-shaped carbon-fiber microelectrodes (CFMEs) are the exposed cut disk face of the fiber itself and the seal region between the carbon fiber and the applied insulating layer. To reduce noise and to fabricate simple, reproducible low-noise CFMEs, we sealed commercially available carbon fibers in pulled glass pipets and then we performed cylindrical etching of the fiber extending beyond the glass sheath, followed by insulation with anodic electrophoretic deposition of paint. The resulting CFMEs had electroactive carbon disks with radii as small as ~0.5 MUm. The noise of such electrodes was minimized by virtue of a design that ensures a good seal between the carbon fiber and its insulation and a reduced diameter of the exposed carbon. In contrast to CFMEs made of conically etched carbon fibers, cylindrically etched CFMEs offer the significant advantage that they can be easily reused: The cylindrically etched region extends over several hundreds of micrometers and, therefore, can be cut back repeatedly to expose a fresh carbon surface of uniform diameter. The low noise and small size of these electrodes make them ideal for the high-sensitivity measurements demanded in studies of single-vesicle transmitter release from secretory cells. Furthermore, the small cross-sectional diameter of the tips allows them to be used in restricted spaces, such as inside the tapering micrometer-diameter tips of melted and pulled glass microcapillaries (e.g., patch pipets). PMID- 21644629 TI - Amperometric gas sensor response times. AB - This work examines the relative importance of diffusion and the electrical time constant of the working electrode on the response time of amperometric gas sensors. The response curve of an electrochemical carbon monoxide sensor to a 200 ppm CO test gas was measured as a function of the added resistance that is in series with the working electrode to increase the time constant. It has been found that the response time increases with additional resistance. The experimental behavior was compared to the values predicted on the basis of Fick's second law of diffusion and to the values expected on the basis of the electrical properties of the sensor. The behavior of the sensor was explained using an equivalent electrical circuit, describing the time constants of the working electrode. Values obtained from this model were further compared to experimental values obtained from electrochemical impedance spectra. It has been found that the response time of the sensors depends on both the rate of diffusion and the electrical time constants of the working electrode. PMID- 21644630 TI - Airtight in Situ Thin-Layer Reflection-Absorption FT-IR Microspectroelectrochemical Cell for the Study of Nonaqueous Systems. AB - An airtight thin-layer reflection FT-IR microspectroelectrochemical cell suitable for the study of both aqueous and nonaqueous electrochemical systems is described. Due to its design, the cell can be easily assembled using disk microelectrodes. This variable-thickness cell can be used for the voltammetric study of both aqueous and nonaqueous systems under semi-infinite diffusion and thin-layer conditions. Dichloromethane solutions of ferrocene were used to test the in situ FT-IR microspectroelectrochemical performance of the cell. The cyclic voltammetric behavior of an oxygen- and water-free CH(2)Cl(2) solution of Ru(3)(CO)(12), under semi-infinite diffusion conditions, provides evidence of the airtightness of this cell. PMID- 21644631 TI - Microscale Synthesis of Fe(PF(3))(5) for Gas Source Mass Spectrometry. AB - Down-scaling of a cocondensation method to produce volatile pentakis(trifluorophosphine)iron(0) for measuring isotope ratios by gas source mass spectrometry is described. Starting from 10 mg of iron, this method allows synthesis of a sufficient amount of Fe(PF(3))(5) that can be used to measure iron content and/or isotope ratios in real-life samples. The advantage over thermal ionisation mass spectrometry is that more precise determinations of ratios/amounts are possible. Reproducibilities for n((54)Fe)/n((56)Fe) and n((57)Fe)/n((56)Fe) are (1-3) * 10(-4), while those obtained by thermal ionization mass spectrometry are at the 10(-3) level. PMID- 21644632 TI - Automated On-Line Isotope Dilution Analysis with ICP-MS Using Sandwich Flow Injection. AB - An automated flow injection (FI) manifold is described to perform the addition of isotopic spikes to aqueous samples on-line with ICP-MS for isotope dilution (ID) analysis. The manifold uses the sandwich technique (with the nested loop approach) to perform an injection of the isotopic spike solution within a sample (or standard) plug, the resulting sample-spike-sample sequence being pushed toward the nebulizer by a 1% HNO(3) carrier. A standard, which must contain one element not present in the spike solution to allow the determination of the dispersion coefficient, must also be used to allow a reverse isotope dilution analysis, as well as corrections for mass discrimination and/or spectroscopic interferences. Indeed, because the signals from the individual isotopes are monitored continuously, only one isotope free of spectroscopic interference is required for elements whose isotopic distribution does not vary in nature (two isotopes are still needed for the other elements), as a correction for the interference can be made by comparison with the signals from the standard. Furthermore, this automated approach makes ID-ICP-MS a faster method and does not require any preliminary analysis of the sample because the concentration profile resulting from FI allows the selection of the best isotopic ratio. It was successfully applied to the determination of Mo in saline water. PMID- 21644633 TI - Characterization of the cationic surfactant induced reversal of electroosmotic flow in capillary electrophoresis. PMID- 21644634 TI - Electrically polarized ion-exchange beds in ion chromatography: ion reflux. AB - All forms of liquid chromatography use eluents that must be prepared and replenished; a widely applied form of ion chromatography (IC) also requires a suppressor and a means for regenerating it. "Ion reflux", as applied to IC, is a new ion-exchange technique where an electrically polarized ion-exchange bed becomes the source of eluent as well as its means of suppression. Using water as the pumped phase, such polarized beds enable the "perpetual" generation and suppression of eluent with little intervention by the user. In one embodiment of ion reflux, continuous eluent generation, ion separation, and continuous suppression are accomplished within a single bed. In another case, where separation is uncoupled from the other two functions, the ion reflux device may be used with existing separators. This paper describes the principles of ion reflux, the advantages and disadvantages of various embodiments, and gives examples of their use in both isocratic and gradient modes of ion separation. These new means for automating eluent generation and suppression should open pathways to new forms of IC instruments and systems. PMID- 21644635 TI - Thermally activated electrostatic injection of solvated ions by a track membrane interfaced vacuum feedthrough. AB - Electromembrane ion sources are considered as potential techniques for direct mass spectrometric sampling from ambient conditions. Interfacing of a time-of flight mass spectrometer by means of a poly(ethylene terephthalate) track membrane requires investigation of the thermally activated processes involved. In this study, we directly attempt substantiating an activation-like performance of such track membrane-mediated interfaces. A number of KCl/glycerol solution samples were tested. A wide range of fixed, externally applied, potential drops was covered. For the charge extraction processes studied, we observe and discuss an activation term, exp(-DeltaF/k(B)T), with a free-energy barrier DeltaF = DeltaF(0) - DeltaF(Phi). The potential drop-dependent DeltaF(Phi), was found to be sensitive to the varying salt concentration. PMID- 21644636 TI - Determination of dissolved metal species by electrospray ionization mass spectrometry. AB - The distribution of metal species in solution was determined using flow injection electrospray ionization mass spectrometry. Complexes formed by selected metal ions with added organic ligands in 50:50 water/acetonitrile and 50:50 water/methanol under acidic, neutral, and basic conditions were detected using electrospray ionization conditions optimized to best represent solution-phase interactions. Metal species containing acetate, nitrate, and solvent molecules predominated in acidic solution but became less abundant at higher pH. Interactions between metal ions and added organic ligands became more selective with increasing pH, showing the expected preference of hard and soft ligands for metal ions of the corresponding type. Species distributions also tended toward larger complexes as pH increased. Overall ion yield was greater for aqueous acetonitrile than for aqueous methanol solutions; however, reduction of copper(II) in aqueous acetonitrile resulted in the detection of copper(I) complexes for certain ligands. Experimental results for copper(II) and 8 hydroxyquinoline in 50:50 water/methanol showed good agreement with aqueous speciation predicted using the thermodynamic equilibrium model MINEQL. Detection of neutral complexes was achieved by protonation, deprotonation, or electrochemical oxidation during electrospray. PMID- 21644637 TI - Minimization of sample discrimination introduced by on-column fracture/electrokinetic injection in capillary electrophoresis. AB - Using on-column fracture/electrokinetic injection for sample introduction in capillary electrophoresis (CE) is thought to be a method of no-discrimination electrokinetic injection. However, in this study we found that significant discrimination was observed when injecting samples dissolved in deionized water with the above method. In addition, the discrimination is reverse to that in conventional electrokinetic injection, that is, the less mobile species are injected in larger quantities than the more mobile components. The reason for these phenomena and approaches to reduce the discrimination were studied. Equivalent circuits were established and used to analyze the discrimination under different conditions through computer simulation. The experimental results showed good agreement with the results of the computer simulations. Finally, an on column fracture/electrokinetic injection method using ramped injection voltage was proposed and shown to be suitable for sample solutions with different conductivity. This method does not cause significant error for practical quantification, even without correcting for the discrimination, because its discrimination is very small. PMID- 21644638 TI - Quantitative Analysis of Anions at ppb/ppt Levels with Capillary Electrophoresis and Conductivity Detection: Enhancement of System Linearity and Precision Using an Internal Standard. AB - Capillary electrophoresis with conductivity detection is evaluated for quantitative analysis of anions at low- to sub-ppb concentration levels in the presence and absence of a conductive sample matrix composed of 2 ppm ammonia and 50 ppb hydrazine. The low-level sensitivity is extended by a transient isotachophoretic stacking procedure. The linear range of the CE system and conductivity detector is graphically evaluated on the basis of absolute and corrected (normalized) chloride and sulfate peak profiles using an ASTM linearity criterion. The influence of random contamination bias from ubiquitous entities of nonsample chloride and sulfate levels introduced by liquid handling, laboratory atmosphere, and bulk chemical residues is quantitatively compared against an internal (contamination) reference ion. PMID- 21644639 TI - Automated particle electrophoresis: modeling and control of adverse chamber surface properties. AB - Electrophoretic analysis of colloidal particles is adversely affected by a host of surface phenomena, including electroosmosis, phase wall wetting, and sample or air bubble adsorption. Neutral, hydrophilic polymer coatings control such phenomena on a variety of surfaces. Poly(ethylene glycol)-poly(ethylene imine) (PEG-PEI) conjugates significantly reduce electroosmosis and positively control adsorption and wetting in the glass sample chambers (5 mm * 3 mm * 1 mm i.d.) employed in a representative commercial electrophoresis apparatus (Coulter DELSA 440). The reduction in electroosmosis (e.g., 80% in 7.5 mM solution at pH 11) was similar to that exhibited by coated 2-mm-i.d. quartz capillaries in a Rank MK I manual apparatus. PEG-PEI coatings significantly reduce electroosmosis over a wide range of pH (2-11) and ionic strength (1-100 mM) and can be stable for weeks under normal laboratory conditions. They greatly enhance ease of operation and accuracy (sample mean electrophoretic mobility +/- SD) of the DELSA 440. The latter results from reduced electroosmosis flow profile gradients near the chamber center-axis stationary levels, where particle mobility is typically measured. Such flow profiles may also be affected by chamber wall surface asymmetries. A hydrodynamic description of electroosmotic fluid flow in rectangular chambers was adapted in order to analyze the propagation of errors due to both nonideal focusing and chamber surface asymmetry. The analysis indicated that the accuracy of rectangular chambered devices may be improved by measuring particle mobility at stationary levels different than chamber center axes. As a result, some rectangular chambers may confer accuracy advantages over cylindrical chambers. PMID- 21644640 TI - Fabricating large arrays of microwells with arbitrary dimensions and filling them using discontinuous dewetting. AB - This paper describes the fabrication of large (up to 45 cm(2)) arrays of microwells, with volumes as small as ~3 fL/well and densities as high as 10(7) wells/cm(2). These arrays of microwells are formed by casting an elastomer, poly(dimethylsiloxane) (PDMS), against "masters" prepared by photolithography; arrays of microwells in other polymers can be formed by using a master consisting of posts in PDMS. A straightforward technique, discontinuous dewetting, allows wells to be filled rapidly (typically on the order of 10(4) wells/s) and uniformly with a wide range of liquids. Several rudimentary strategies for addressing microwells are investigated, including electroosmotic pumping and gaseous diffusion. PMID- 21644641 TI - Sampling and determination of formaldehyde using solid-phase microextraction with on-fiber derivatization. AB - Gaseous formaldehyde is sampled by derivatization with o-(2,3,4,5,6 pentafluorobenzyl)hydroxylamine hydrochloride (PFBHA) adsorbed onto poly (dimethylsiloxane)/divinylbenzene solid-phase microextraction fibers. The product of the reaction is an oxime which is thermally very stable and insensitive to light. The oxime can be analyzed by gas chromatography with flame ionization detection and other detectors. Loading PFBHA on the fiber is by room-temperature headspace extraction from aqueous solutions of PFBHA. The process of loading and desorption of unreacted PFBHA, and oxime formed, is both highly reproducible and reversible, with more than 200 loading, sampling, and analysis steps possible with one fiber. The standard formaldehyde gas concentrations studied ranged from 15 to 3200 ppbv with sampling times from 10 s to 12 min. Quantification can be achieved via interpolation from calibration curves of area counts as a function of formaldehyde concentration for a fixed sampling time. Sampling for 10 s yields a method detection limit of 40 ppbv and at 300 s the method detection limit is 4.6 ppbv. This is equal to or better than all other conventional grab sampling methods for gaseous formaldehyde employing sampling trains or passive sampling techniques. Alternatively, gaseous formaldehyde can be quantified with an empirically established apparent first-order rate constant (0.0030 ng/(ppbv s) at 25 degrees C) for the reaction between sorbed PFBHA and gaseous formaldehyde. This first-order rate constant allows for quantitative analyses without a calibration curve, only requiring detector calibration with the oxime. This new method was used for the headspace sampling of air known to contain formaldehyde, as well as other carbonyl compounds, and from various matrixes such as cosmetics and building products. PMID- 21644642 TI - Supercritical transitiometry of polymers. AB - Employing supercritical fluids (SCFs) during polymers processing allows the unusual properties of SCFs to be exploited for making polymer products that cannot be obtained by other means. A new supercritical transitiometer has been constructed to permit study of the interactions of SCFs with polymers during processing under well-defined conditions of temperature and pressure. The supercritical transitiometer allows pressure to be exerted by either a supercritical fluid or a neutral medium and enables simultaneous determination of four basic parameters of a transition, i.e., p, T, Delta(tr)H and Delta(tr)V. This permits determination of the SCF effect on modification of the polymer structure at a given pressure and temperature and defines conditions to allow reproducible preparation of new polymer structures. Study of a semicrystalline polyethylene by this method has defined conditions for preparation of new microfoamed phases with good mechanical properties. The low densities and microporous structures of the new materials may make them useful for applications in medicine, pharmacy, or the food industry, for example. PMID- 21644643 TI - Surface Plasmon Resonance-Laser Desorption/Ionization-Time-of-Flight Mass Spectrometry. AB - The laser desorption/ionization (LDI) process is investigated under surface plasmon resonance (SPR) conditions using time-of-flight mass spectrometry (TOFMS). We demonstrate that LDI-TOFMS at the SPR angle requires a lower minimum laser fluence for the production of silver monomer and cluster cations from ablation of a thin silver film substrate. In the LDI of gramicidin S deposited on a thin silver film substrate, the largest intensity for the molecular cation peak occurs when the laser light is incident on the substrate at a specific SPR angle. These results fully confirm SPR enhancement of the LDI process. The capability to perform SPR-LDI on a larger molecular weight analyte (1141 amu for gramicidin S) represents a new milestone beyond the previous achievement with rhodamine B (479 amu). A better understanding of the SPR mechanism is gained with respect to the substrate metals (silver vs aluminum), desorption (microscopic vs mesoscopic), and ionization (chemical vs multiphoton). These findings may be useful in the future design of SPR-LDI techniques for better TOFMS analysis of higher mass biomolecules. PMID- 21644644 TI - Outlier detection in multivariate analytical chemical data. AB - The unreliability of multivariate outlier detection techniques such as Mahalanobis distance and hat matrix leverage has been known in the statistical community for well over a decade. However, only within the past few years has a serious effort been made to introduce robust methods for the detection of multivariate outliers into the chemical literature. Techniques such as the minimum volume ellipsoid (MVE), multivariate trimming (MVT), and M-estimators (e.g., PROP), and others similar to them, such as the minimum covariance determinant (MCD), rely upon algorithms that are difficult to program and may require significant processing times. While MCD and MVE have been shown to be statistically sound, we found MVT unreliable due to the method's use of the Mahalanobis distance measure in its initial step. We examined the performance of MCD and MVT on selected data sets and in simulations and compared the results with two methods of our own devising. Both the proposed resampling by the half means method and the smallest half-volume method are simple to use, are conceptually clear, and provide results superior to MVT and the current best performing technique, MCD. Either proposed method is recommended for the detection of multiple outliers in multivariate data. PMID- 21644645 TI - Surface-enhanced Raman scattering on uniform transition-metal films: toward a versatile adsorbate vibrational strategy for solid-nonvacuum interfaces? AB - Procedures are outlined for the electrodeposition of ultrathin films of Pt-group transition metals onto gold that provide intense surface-enhanced Raman scattering (SERS) for adsorbates bound to the overlayers yet (unlike earlier reports) are sufficiently "pinhole-free" to avoid significant spectral and chemical interferences from the underlying substrate. Constant-current electrodeposition of Pd, Rh, Pt, and Ir from perchloric acid and/or phosphate electrolytes yields essentially layer-by-layer growth, so that near-ideal pinhole free electrochemical and spectral characteristics are achieved for film thickness of ~2 monolayers or more. The desired film uniformity is diagnosed from the voltammetric oxide formation-removal behavior and, especially, from the absence of the characteristic C-O stretching (nu(CO)) SERS band at 2110-2120 cm(-)(1) due to CO binding to Au surface sites. Carbon monoxide is also employed as a surface environment-sensitive adsorbate to probe the electrochemical SERS characteristics as a function of the transition-metal film thickness. The Raman enhancement was observed to decrease by 2-fold every 10-20 A or so, exhibiting a "spacer distance" dependence that is weaker than (but functionally similar to) recently reported organic insulator films. The practical value of the present films for obtaining rich vibrational spectra for diverse adsorbates on transition metals is pointed out and briefly illustrated for benzonitrile on a Pt film electrode. The more general promise of this overlayer film SERS strategy as a versatile vibrational technique for characterizing other types of chemically important surface materials is also noted. PMID- 21644646 TI - A device for sampling and determination of total particulate mercury in ambient air. AB - A miniaturized device, which serves as both particulate trap and pyrolyzer for airborne particulate mercury species, is described. It has been used in combination with amalgamation/thermal desorption/cold vapor atomic fluorescence spectrometry detection for the determination of total particulate mercury (TPM) associated with atmospheric aerosols. A standard reference material (SRM 1633b, NIST) has been used for validating of the pyrolysis technique, and a relative error smaller than 3% has been obtained. Contrary to most methods currently employed, this new technique does not require any sample preparation (e.g., extraction/digestion), no manual sample transfer or sample handling, and no addition of chemicals or reagents. Hence the risk of contamination is low. The time for complete analysis is less than 10 min per sample. The concentrations of TPM determined in metropolitan Toronto ranged from 3 to 91 pg m(-)(3) with standard deviations of <+/-2 pg m(-)(3) for simultaneous sets of four samples. These atmospheric TPM concentration values fall within the range reported in the literature. Good agreement was obtained by the three methods compared in a field study at Ny-Alesund (78 degrees 54'N, 11 degrees 53'E), Svalbard. The elevated values of TPM concentrations obtained using the method developed in this work may arise from the Arctic springtime conversion of atmospheric mercury from gas-phase to particulate-phase Hg species. PMID- 21644647 TI - Fast aerosol analysis by fourier transform imaging fluorescence microscopy. AB - Fourier transform imaging spectroscopy was combined with fluorescence microscopy and a cooled CCD detector for fast analysis of aerosols contaminated with polycyclic aromatic hydrocarbons (PAHs). Aerosols were collected on glass fiber filters and inspected, for the first time, by this imaging technique, which provides a full fluorescence spectrum at each pixel. Mapping of PAH contamination was carried out and used for identification and quantification of the compounds. Quantification limits (based on 95% confidence intervals of calibration plots) in the 10 ng cm(-)(2) range on filter are reported, which corresponds to 20 ng m( )(3) in air, integrated in 1 min. The absolute detection limit (on filter) is estimated as low as 0.25 pg, corresponding to an air concentration of 0.5 pg m( )(3), integrated in 1 min. The method is examined for analysis of monocomponent contamination and for simple mixtures. After a proper automation, this method has the potential to provide in situ and on-line results regarding particulate airborne PAH contaminations. PMID- 21644648 TI - UV-Visible Spectroscopic Measurement of Solubilities in Supercritical CO(2) Using High-Pressure Fiber-Optic Cells. AB - The design and construction of a microscale, fiber-optics-based system for the measurement of solubilities in supercritical CO(2) by UV-visible spectroscopy is described. This system consists of three high-pressure fiber-optic cells, with path lengths ranging from 38 MUm to 1 cm, constructed from standard (1)/(16)-in. stainless steel fittings and silica fibers. It is capable of withstanding pressures in excess of 300 atm, and spectra over the entire UV-visible range (200 900 nm) can be obtained. Use of three cells with different path lengths enables compounds of high or low solubility to be measured over a concentration range of several orders of magnitude. The solubility of a uranium complex, UO(2)(tta)(2).TBP, in supercritical CO(2) at 40 degrees C and over the pressure range 100-325 atm was determined, and it was found to be possible to attain solubilities in excess of 10(-)(2) M for metal species in unmodified supercritical CO(2). Also, the small volume of this system allows solubilities to be measured with relatively small amounts of compounds. PMID- 21644649 TI - Focused microwave-assisted soxhlet: an advantageous tool for sample extraction. AB - A new type of microwave-assisted Soxhlet is here reported. The device uses conventional Soxhlet glassware for solid sample extraction and a focused microwave digester for irradiation of the sample cartridge at the required intervals while the fresh solvent (condensed vapors from the distillation flask) drips on and passes through the solid sample. In this way breaking of the analyte matrix bonds is facilitated by application of the appropriate energy. The new approach has been checked in a comparative study by its application to the extraction of analyte families of different polarity (namely, alkanes, polyaromatic hydrocarbons, and herbicides) from the same soil matrix using dichloromethane as extractant. The reduction of the extraction times (from 8 h to 50-60 min, depending on the polarity of the analytes) with efficiency similar to or even higher than that afforded by the conventional Soxhlet technique supports the suitability of the new approach. In addition, recycling of the solvent during extract preconcentration enables minimal environmental contamination to be achieved. PMID- 21644650 TI - Laser desorption-membrane introduction mass spectrometry. AB - In this paper we describe the first use of laser desorption in conjunction with membrane introduction mass spectrometry (MIMS). In this technique, a low-powered carbon dioxide laser is used to irradiate the low-pressure (vacuum) side of a silicone membrane during a typical MIMS analysis of an aqueous solution. The absorption of laser energy results in direct membrane heating and rapid desorption of permeate molecules. This improves both the sensitivity and response times of MIMS when analyzing compounds having high molecular weight and low volatility. Two simple interfaces are described for performing laser desorption inside and outside the vacuum manifold of a GCQ ion trap mass spectrometer. Together with flow injection (FI) sample introduction, we demonstrate direct on line monitoring of aqueous solutions of high boiling point (200-530 degrees C) polynuclear aromatic hydrocarbons such as naphthalene, anthracene, pyrene, benzo[b]fluoranthene, and indeno[123-cd]pyrene. PMID- 21644651 TI - A Simple Method for Examining the Electrochemistry of Metalloporphyrins and Other Hydrophobic Reactants in Thin Layers of Organic Solvents Interposed between Graphite Electrodes and Aqueous Solutions. AB - A method is described for the preparation of stable, adherent, thin layers of organic solvents interposed between the surfaces of graphite electrodes and aqueous supporting electrolytes in which they are immersed. The electrochemistry of reactants dissolved in the thin layers is examined and utilized to evaluate surface coverages and formal potentials for molecules, such as cobalt tetraphenylporphyrins, that do not exhibit useful electrochemical responses when adsorbed on graphite. The thin layers of organic solvent can be used to concentrate analytes by extracting them from dilute aqueous solutions to produce enhanced sensitivity in electroanalytical applications. Electron transfer across the liquid/liquid interface created by the presence of the thin layer of immiscible organic solvent is also demonstrated. PMID- 21644652 TI - A Fill-and-Flow Biosensor. AB - An alternative design for an amperometric biosensor for lactate is described in which the analyte flows through a rectangular duct without the aid of external pumping or a flow system. With this deviation from the traditional "stacked layers" of the biosensor geometry, the biorecognition matrix is located upstream of the detector electrode rather than directly over the electrode; this general design is reminiscent of the "dipstick" technology of the home test kits, but in the latter case the "flow" is a function of the nature of the wicking material rather than the dimensions of the channel. A droplet of analyte, l-lactate, is placed in the inlet well of the channel which flows through the cell and reacts with an immobilized lactate oxidase surface matrix zone, and the coproduct, hydrogen peroxide, is swept downstream to the monitoring electrode, where it is detected. The flow through the channel biosensor is shown to be laminar, and the biosensor response varies with the rate of flow of the analyte, which could be controlled by the outlet porosity; the sensitivity was observed to increase, but the dynamic range decreased with an increase in flow rate until a threshold loading, where the biosensor response became independent of the enzyme loading. PMID- 21644653 TI - Anthraquinonedisulfonate electrochemistry: a comparison of glassy carbon, hydrogenated glassy carbon, highly oriented pyrolytic graphite, and diamond electrodes. AB - The electrochemistry of anthraquinone-2,6-disulfonate (2,6-AQDS) at glassy carbon (GC), hydrogenated glassy carbon (HGC), the basal plane of highly oriented pyrolytic graphite (HOPG), and boron-doped diamond was investigated by cyclic voltammetry and chronocoulometry. Quantitative determination of the surface coverage and qualitative assessment of the physisorption strength of 2,6-AQDS adsorption on each of these electrodes were done. The diamond and HGC surfaces are nonpolar and relatively oxygen-free, with the surface carbon atoms terminated by hydrogen. The polar 2,6-AQDS does not adsorb on these surfaces, and the electrolysis proceeds by a diffusion-controlled reaction. Conversely, the GC and HOPG surfaces are polar, with the exposed defect sites terminated by carbon oxygen functionalities. 2,6-AQDS strongly physisorbs on both of these surfaces at near monolayer or greater coverages, such that the electrolysis proceeds through a surface-confined state. Less than 40% of the initial surface coverage can be removed by rinsing and solution replacement, reflective of strong physisorption. The results show the important role of the surface carbon-oxygen functionalities in promoting strong dipole-dipole and ion-dipole interactions with polar and ionic molecules such as 2,6-AQDS. The results also support the theory that diamond electrodes may be less subject to fouling by polar adsorbates, as compared to GC, leading to improved response stability in electroanalytical measurements. The relationship between the 2,6-AQDS surface coverage, the double layer capacitance, and the heterogeneous electron-transfer rate constant for Fe(CN)(6)(3)(-)(/4)(-) for these four carbon electrodes is presented. PMID- 21644654 TI - Voltammetry at micropipet electrodes. AB - The use of micropipet electrodes for quantitative voltammetric measurements of ion-transfer (IT) and electron-transfer (ET) reactions at the interface between two immiscible electrolyte solutions (ITIES) requires knowledge of geometry of the liquid interface. The shape of the meniscus formed at the pipet tip was studied in situ by video microscopy under controlled pressure. The shape of the interface can be changed from a complete sphere to a concave spherical cap by varying the pressure applied to the pipet, and the diffusion current to the pipet changes accordingly. With no external pressure applied, the water/organic interface turned out to be flat, and the voltammetric response of a pipet must follow the well-known theory for a microdisk electrode. The large deviations from this theory observed previously can be attributed to a small amount of the filling aqueous solution which escapes from the pipet and forms a thin layer on its outer wall. This effect can be eliminated by making the outer pipet wall hydrophobic. Procedures have been developed for independent silanization of the inner and outer walls of the pipet. Pipets with a silanized inner wall can be filled with an organic solvent (e.g., 1,2-dichloroethane) and be used for voltammetric measurements in aqueous solutions. Another mode of voltammetry is based on trapping of a thin layer of organic solvent in the narrow shaft of a pipet between the filling solution and the aqueous outer phase. This arrangement is potentially useful for electrochemical catalysis and sensor applications. PMID- 21644655 TI - Absorption Anomalies in Ratio and Subtraction FT-IR Spectroscopy. AB - Subtraction and ratioing of strong absorption bands in Fourier transform infrared (FT-IR) spectroscopy produces anomalous absorption errors. One source of error is the instability in the wavenumber scale of the FT-IR spectra. The possible causes of this error are explored. The thermal expansion and contraction of the cavity of the HeNe reference laser from a typical commercial instrument was found to produce changes in the laser wavenumber of +/-0.034 cm(-)(1). Changes of this size are shown to introduce errors into the wavenumber scales of FT-IR spectra which are sufficient to produce the observed anomalies. The dependence of the error on instrumental and spectroscopic parameters is explored. Solutions to the problem are proposed. PMID- 21644656 TI - A drift correction procedure. AB - A procedure is introduced that can mitigate the deleterious effect of low frequency noise [Formula: see text] often termed drift [Formula: see text] on the precision of an analytical experiment. This procedure offers several performance benefits over traditional designs based on the periodic measurement of standards to diagnose and correct for variation in instrument response. Using repeated measurements of every sample as a drift diagnostic, as opposed to requiring the periodic measurement of any given sample or standard, the analyst can better budget the measurement time to be devoted to each sample, distributing it to optimize the uncertainty of the analytical result. The drift is diagnosed from the repeated measurements, a model of the instrument response drift is constructed, and the data are corrected to a "drift-free" condition. This drift free condition allows data to be accumulated over long periods of time with little or no loss in precision due to drift. More than 10-fold precision enhancements of analytical atomic emission results have been observed, with no statistically significant effects on the means. The procedure is described, performance data are presented, and matters regarding the procedure are discussed. PMID- 21644657 TI - Laser-induced fluorescence of perylene in a microparticle suspension environment. AB - Laser-induced fluorescence (LIF) is one of the most sensitive techniques for analysis of traces of polycyclic aromatic compounds in liquids. Application of this method to on-line monitoring requires solution of problems related to the presence of particulate materials. Thus, understanding the analytical effects associated with the suspension of microparticles is of considerable importance for both environmental and industrial applications. Here, LIF of perylene in acetonitrile solutions was studied for different light-scattering/-absorbing microparticle matrixes. With increasing suspension mass of efficiently light absorbing black-colored particles, the intensity of the associated LIF signals was found to obey an apparently exponential decrease. Their white-colored counterparts, however, have initially produced a sizable, ~20%, signal increase of the respective LIF responses. An exponential decrease then became predominant, too. A model that explains the observations in terms of absorption and scattering coefficients is developed and examined. The quantitative effect of the particulate mass is correctly reproduced by the model, as well as the laser wavelength dependence. A possible application of a calibrating algorithm is addressed. PMID- 21644658 TI - Characterization of the sources of variation affecting near-infrared spectroscopy using chemometric methods. AB - A rapid assessment of product quality can often be made using a combination of near-infrared spectroscopy (NIR) and multivariate calibration. The robustness of such a method is determined by the sensitivity of the multivariate calibration model to variations in the spectral data. An approach is described that uses a combination of experimental design methodology and principal component analysis to identify the main sources of variation in the spectra and to estimate their influence on the quantitative predictions. This is accomplished by comparing variations in a set of measured, replicate spectra to spectra with simulated variations. The approach was applied to the hydroxyl number determination of polyols by NIR spectroscopy and partial least-squares calibration. The results indicated that the most significant sources of variation were due to a variable cell path length and a variable curved background. Correction for these errors resulted in a 58% reduction in the standard deviation of the hydroxyl number predictions, indicating that a substantial improvement in the method precision is possible. PMID- 21644659 TI - Identification, Composition, and Asymmetric Formation Mechanism of Glycidyl Methacrylate/Butyl Methacrylate Copolymers up to 7000 Da from Electrospray Ionization Ultrahigh-Resolution Fourier Transform Ion Cyclotron Resonance Mass Spectrometry. AB - Glycidyl methacrylate (GMA) and butyl methacrylate (BMA) have the same nominal mass (142 Da) but differ in exact mass by 0.036 Da (CH(4) vs O). Therefore, copolymers formed from the two isobaric monomers exhibit a characteristic isobaric distribution due to different monomer compositions. Here, we show that electrospray ionization FT-ICR mass spectrometry at 9.4 T resolves the isobaric components of copolymers as large as 7000 Da with a resolving power (m/Deltam(50%)) of ~500 000 in a gel permeation chromatography fractionated polymer sample. That resolution provides for complete and unequivocal component analysis of such copolymers of the size used for high solid content automobile coatings. All five possible copolymer products predicted by the polymerization mechanism are resolved and identified in the mass spectrum. Two of those polymer series (each with saturated end group) were previously unresolved by mass spectrometry because they differ in mass from the two other unsaturated products by only 0.0089 Da. Finally, analysis of the asymmetrical isobaric distribution for the copolymer n-mers, (GMA)(m)(BMA)(n)(-)(m), 0<= m <= n, in which species with adjacent values of m differ from each other in mass by 36 mDa (i.e., the mass difference, CH(4) vs O, between GMA and BMA) proves that GMA is less reactive than BMA in the polymerization process. PMID- 21644660 TI - High-throughput flow injection analysis mass spectroscopy with networked delivery of color-rendered results. 2. Three-dimensional spectral mapping of 96-well combinatorial chemistry racks. AB - For the last two years, the mass spectroscopy section of the Novartis Pharma Research Core Technology group has analyzed tens of thousands of multiple parallel synthesis samples from the Novartis Pharma Combinatorial Chemistry program, using an in-house developed automated high-throughput flow injection analysis electrospray ionization mass spectroscopy system. The electrospray spectra of these samples reflect the many structures present after the cleavage step from the solid support. The overall success of the sequential synthesis is mirrored in the purity of the expected end product, but the partial success of individual synthesis steps is evident in the impurities in the mass spectrum. However this latter reaction information, which is of considerable utility to the combinatorial chemist, is effectively hidden from view by the very large number of analyzed samples. This information is now revealed at the workbench of the combinatorial chemist by a novel three-dimensional display of each rack's complete mass spectral ion current using the in-house RackViewer Visual Basic application. Colorization of "forbidden loss" and "forbidden gas-adduct" zones, normalization to expected monoisotopic molecular weight, colorization of ionization intensity, and sorting by row or column were used in combination to highlight systematic patterns in the mass spectroscopy data. PMID- 21644661 TI - A method for quantitatively differentiating crude natural extracts using high performance liquid chromatography-electrospray mass spectrometry. AB - This paper describes a method for quantitatively differentiating crude natural extracts using high-performance liquid chromatography-electrospray mass spectrometry (HPLC-ESI-MS). The method involves performing an HPLC-MS analysis using standard reversed-phase C18 gradient separation on the crude extract. The HPLC system used in this study was a dual-column system designed to optimize throughput. Using image analysis techniques, the data are reduced to a list containing the m/z value and retention time of each ion. The ion lists are then compared in a pairwise fashion to compute a sample similarity index between two samples. The similarity index is based on the number of ions common to both and is scaled from 0 to 1. Extract controls were analyzed throughout a run of 88 unknown fungal extracts. The controls provided information about column and spectrometer stability and overall sensitivity. Pairwise comparison of all control samples indicates that the similarity index is high (0.8) for replicate samples. Comparison between the unknown extract samples produces a distribution of similarities ranging from replicates (0.8) to very dissimilar (0.1). This information can be used to judge the chemical diversity of natural extract samples, which is one approach to determining the quality of libraries being used for drug discovery via high-throughput screening. PMID- 21644662 TI - Higher order equilibria and their effect on analyte migration behavior in capillary electrophoresis. AB - This paper presents a quantitative investigation into the effect of analyte additive interactions on analyte migration behavior in capillary electrophoresis (CE) when both 1:1 and 1:2 stoichiometries are present. Equations based on the individual capacity factors for each interaction are derived to account for the effect of both first- and second-order equilibria. The analyte migration behavior is described using these equations with a full account of how the microscopic equilibrium constants and microscopic mobilities are combined to give the macroscopic values. The binding isotherms of interactions with both 1:1 and 1:2 stoichiometries are compared with those of a 1:1 stoichiometry. 4,4'-Biphenol and 4-phenylphenol were chosen as analytes that undergo complexation with one and two hydroxypropyl-beta-cyclodextrin (HP-beta-CD) molecules; phenol was used as an analyte that interacts with only one HP-beta-CD molecule. The process of calculating higher order equilibrium constants and complex mobilities from the binding isotherms is demonstrated. The effects of experimental conditions, such as the additive concentration range and the number of data points, on the error in the calculated constants and the ability of the equations to accurately describe the experimental data are discussed. A comparison of the linear transformations of the binding isotherm with respect to their ability to detect higher order equilibria is made, and the advantage of using the capacity factor in CE is illustrated. PMID- 21644663 TI - Separation of explosives using capillary electrochromatography. AB - The identification of explosives and their degradation products is important in forensic and environmental applications. Complete separation of these structurally similar compounds using reversed-phase liquid chromatography has proven to be a challenge. Here we present a demonstration of the use of capillary electrochromatography on the separation of a series of 14 nitroaromatic and nitramine explosive compounds. A separation with baseline resolution is achieved for all of the compounds in under 7 min, featuring efficiencies of over 500 000 theoretical plates/m. Using more aggressive running conditions, 13 of the 14 compounds are separated in under 2 min. PMID- 21644664 TI - On-line coupling of capillary electrochromatography, capillary electrophoresis, and capillary HPLC with nuclear magnetic resonance spectroscopy. AB - A novel capillary NMR coupling configuration, which offers the possibility of combining capillary zone electrophoresis (CZE), capillary HPLC (CHPLC), and for the first time capillary electrochromatography (CEC) with nuclear magnetic resonance (NMR), has been developed. The hyphenated technique has a great potential for the analysis of chemical, pharmaceutical, biological, and environmental samples. The versatile system allows facile changes between these three different separation methods. A special NMR capillary containing an enlarged detection cell suitable for on-line NMR detection and measurements under high voltage has been designed. The acquisition of 1D and 2D NMR spectra in stopped-flow experiments is also possible. CHPLC NMR has been performed with samples of hop bitter acids. The identification and structure elucidation of humulones and isohumulones by on-line and stopped-flow spectra has been demonstrated. The suitability of the configuration for electrophoretic methods has been investigated by the application of CZE and CEC NMR to model systems. PMID- 21644665 TI - Isotopic separation of [(14)n]- and [(15)n]aniline by capillary electrophoresis using surfactant- controlled reversed electroosmotic flow. AB - Separation of isotopically labeled [(14)N]- and [(15)N]aniline was achieved using capillary electrophoresis based on the isotopic effect on pK(a). The effects of the buffer co-ion, pH, and electroosmotic mobility on the resolution are investigated in this paper. Electroosmotic flow (EOF) was controlled using the zwitterionic surfactant Rewoteric AM CAS U as buffer additive. The resultant EOF was anodic (reversed) and low in magnitude (0.6 * 10(-)(4) cm(2)/(V.s)). The resolution of [(14)N]- and [(15)N]aniline was 1.22. Addition of a cationic surfactant, cetyltrimethylammonium bromide, to the zwitterionic surfactant increased the magnitude of the anodic EOF. This EOF improved the resolution to 1.33 based on mobility counterbalance. PMID- 21644666 TI - Solvent-programmed microchip open-channel electrochromatography. AB - Open-channel electrochromatography in combination with solvent programming is demonstrated using a microchip device. Channel walls were coated with octadecylsilanes at ambient temperatures, yielding stationary phases for chromatographic separations of neutral dyes. The electroosmotic flow after coating was sufficient to ensure transport of all species and on-chip mixing of isocratic and gradient elution conditions with acetonitrile-buffer mixtures. Chips having different channel depths between 10.2 and 2.9 MUm were evaluated for performance, and van Deemter plots were established. Channel depths of about 5 MUm were found to be a good compromise between efficiency and ease of operation. Isocratic and gradient elution conditions were easily established and manipulated by computer-controlled application of voltages to the terminals of the microchip. Linear gradients with different slopes, start times, duration times, and start percentages of organic modifier proved to be powerful tools to tune selectivity and analysis time for the separation of a test mixture. Even very steep gradients still produced excellent efficiencies. Together with fast reconditioning times, complete runs could be finished in under 60 s. PMID- 21644667 TI - Improvements in polymer characterization by size-exclusion chromatography and liquid chromatography at the critical condition by using enhanced-fluidity liquid mobile phases with packed capillary columns. AB - Microscale chromatography has found numerous applications in liquid chromatography. The combination of enhanced-fluidity liquid mobile phases with packed-capillary LC is evaluated for polymer characterization using size exclusion chromatography (SEC) and liquid chromatography at the critical condition (LCCC) phase. Separations of polystyrene polymers and copolymers are completed using liquid chromatography at the critical condition. The critical conditions of polystyrene polymers were approached by changing the concentration of CO(2) in the mixture combined with temperature and pressure variation. Because the solvent strength of enhanced-fluidity liquid mixtures is affected by temperature and pressure variation, the solvent strength could be fine-tuned to accurately find the critical condition. Long packed capillaries could be used in this application because the enhanced-fluidity mobile phases have low viscosities. High efficiencies resulted. The performance of packed-capillary and analytical-scale analytical columns containing the same packing material was compared for a challenging separation at the critical condition. PMID- 21644668 TI - Quantitative Analysis of Trace Moisture in N(2) and NH(3) Gases with Dual-Cell Near-Infrared Diode Laser Absorption Spectroscopy. AB - This paper demonstrates our optical measurement system based on near-infrared tunable diode laser absorption spectrometry and reports the results of trace moisture determination in nitrogen and ammonia gases. A near-infrared InGaAsP distributed feedback diode laser operating at room temperature was employed as the optical source. We used a dual-cell detection strategy to cancel common mode noise from the diode laser and remove the effect of the residual moisture absorption in the beam path outside the sample cell. We also used this method to successfully eliminate the interfering absorption of matrix gas molecules such as NH(3). The detection limit of H(2)O absorption of 4 ppb in nitrogen and 12 ppb in ammonia was obtained using a single-pass absorption cell of only 92 cm in length and the average results of 10 scan measurements. This system has characteristics of both the high sensitivity and capability of in situ and real-time measurement. PMID- 21644670 TI - Other/miscellaneous. PMID- 21644669 TI - Stereochemical Differentiation of Mannose, Glucose, Galactose, and Talose Using Zinc(II) Diethylenetriamine and ESI-Ion Trap Mass Spectrometry. PMID- 21644671 TI - High-resolution mass spectrometry with a multiple pass quadrupole mass analyzer. AB - The peak shape narrows and the resolution improves if the ions are simply reflected back and forth through a conventional quadrupole mass analyzer. CO(+) and N(2)(+) at m/z = 28 are separated to 50% valley with half of the original signal remaining. These two ions can be resolved to baseline (m/Deltam) = 5000 with 1% of the original signal remaining. PMID- 21644672 TI - A miniature cylindrical quadrupole ion trap: simulation and experiment. AB - A cylindrical quadrupole ion trap (r(0) = 2.5 mm, z(0) = 2.88 mm, ~(1)/(64) of the volume of commercial hyperbolic ion traps) has been constructed, its geometry optimized, and its performance examined in the mass-selective instability scan mode. Spectra of ionized perfluorotributylamine and o-dichlorobenzene show a resolution (m/Deltam, 50% valley definition) of ~100. The instrument has been coupled to a membrane introduction system to test its applicability for on-line reaction monitoring and to determine detection limits. Simulations using the ion trap simulation program are used to explore the effects of geometry on performance and to validate the experimental results. PMID- 21644673 TI - Development of a digital micromirror spectrometer for analytical atomic spectrometry. AB - A digital micromirror device (DMD) has been incorporated into a novel spectrometer for use in analytical atomic spectrometry. The device can be taken from a commercial computer projector. A protective glass window covering the DMD chip limits the viewable wavelengths to the visible range. The DMD is used to project an image of the light source onto the exit plane of a flat-field spectrograph. A single photomultiplier tube is used for detection. The high switching rate of the micromirrors (15 MUs) enables rapid full-spectrum capture, wavelength-modulation, source-modulation, fast narrow-wavelength window scans, and rapid-wavelength "jumping." Calcium, sodium, and potassium have been determined in several standard reference materials (tomato leaves, bovine liver, rice flour, total diet) by flame atomic absorption and emission spectrometry. Absorption sensitivities for each element are near the 0.02 MUg/mL level, and detection limits for both absorption and emission are near the 0.01 MUg/mL level. Elemental recoveries were within 10% of certified values for most reference materials. PMID- 21644674 TI - Alkyl chain conformation of octadecylsilane stationary phases by Raman spectroscopy. 1. Temperature dependence. AB - Raman spectroscopy is used for the first time to probe the effect of temperature on the conformational order of polymeric and monomeric octadecylsilane stationary phases. Spectral data in the nu(C-C) and nu(C-H) regions are interpreted in terms of alkyl chain conformational state and its dependence on temperature. In contrast to the liquidlike disordered state characteristic of these stationary phases at room temperature, at liquid N(2) temperatures, the alkyl chains exist in a more ordered state with a residual level of gauche conformational defects. Systematic studies between -15 and 95 degrees C reveal more subtle changes in conformational order as ascertained from empirical spectral indicators including the intensity ratios I[nu(a)(CH(2))]/I[nu(s)(CH(2))] and I[nu(C-C)(T)]/I[nu(C C)(G)]. Plots of these ratios as a function of temperature reveal two distinct regimes of behavior. By extrapolating the linear regions of these plots, a surface "phase transition" temperature of ~20 degrees C for both surface confined octadecylsilane stationary phases is estimated that represents subtle changes in alkyl chain conformational order from a more ordered phase to a slightly more disordered phase. The similarity in behavior between the polymeric and monomeric octadecylsilane stationary phases is interpreted as evidence for similar interchain spacing of the alkylsilanes on these silica surfaces. PMID- 21644675 TI - Method performance and validation for quantitative analysis by (1)h and (31)p NMR spectroscopy. Applications to analytical standards and agricultural chemicals. AB - Nuclear magnetic resonance (NMR) can be used to provide an independent and intrinsically reliable determination of chemical purity. Unlike chromatography, it is possible to employ a universal reference standard as an internal standard for the majority of chemical products assayed by quantitative NMR (QNMR). This is possible because the NMR response can be made the same for all chemical components, including the internal standard, by optimizing certain instrumental parameters. Experiments were performed to validate the quantitative NMR method described in this paper for the analysis of organic chemicals. Experimental precision, accuracy, specificity, linearity, limits of detection and quantitation, and ruggedness were systematically addressed, and system suitability criteria were established. The level of the major chemical ingredient can be determined with accuracy and precision significantly better than 1%, and impurities may be quantified at the 0.1% level or below. Thus, QNMR rivals chromatography in sensitivity, speed, precision, and accuracy, while avoiding the need for a reference standard for each analyte. Examples are given of (1)H and (31)P NMR used for quantitative analysis of agricultural chemicals, and a method for characterization of analytical standards is presented. PMID- 21644676 TI - Electrospray ionization high-resolution ion mobility spectrometry-mass spectrometry. AB - A hybrid atmospheric pressure ion mobility spectrometer is described which exhibits resolving power approaching the diffusion limit for singly and multiply charged ions (over 200 for the most favorable case). Using an electrospray ionization source and a downstream quadrupole mass spectrometer with electron multiplier as detector, this ESI-IMS-MS instrument demonstrates the potential of IMS for rapid analytical separations with a resolving power similar to liquid chromatography. The first measurements of gas-phase mobility spectra of mass identified multiply charged ions migrating at atmospheric pressure are reported. These spectra confirm that collision cross sections are strongly affected by charge state. Baseline separations of multiply charged states of cytochrome c and ubiquitin demonstrate the improved resolving power of this instrument compared with previous atmospheric pressure ion mobility spectrometers. The effects of electric potential, initial pulse duration, ion-molecule reactions, ion desolvation, Coulombic repulsion, electric field homogeneity, ion collection, and charge on the resolving power of this ion mobility spectrometer are discussed. PMID- 21644677 TI - Column Selectivity Programming and Fast Temperature Programming for High-Speed GC Analysis of Purgeable Organic Compounds. AB - High-speed gas chromatograms are obtained by the use of relatively short lengths of capillary column operated at relatively large carrier gas flow rates. This approach is difficult for more complex mixtures because of the reduced peak capacity available with shorter columns. A solution to this problem is the use of tunable column ensembles consisting of the series (tandem) combination of a polar and a nonpolar column. By adjusting the pressure at the junction point between the columns, the selectivity of the ensemble can be adjusted within the limits imposed by the individual columns. For mixtures representing a relatively large boiling point range and containing more than ~20 components, high-speed, isothermal separations are less effective. These limitations are significantly reduced by combining fast temperature programming with selectivity programming. Selectivity programming is obtained by changing the pressure at the column junction point one or more times during the course of an analysis. In the work described here, the column ensemble temperature and the junction pressure are initially set to give a high-quality separation of the earliest eluting components. After these components have eluted, a linear temperature ramp of ~35 degrees C/min is initiated. As the temperature increases, the pressure is adjusted to change the selectivity and thus facilitate the separation of groups of components as they migrate through the column ensemble. Using this approach, a mixture of 30 purgeable organic compounds is separated in less than 2.5 min. PMID- 21644678 TI - Combinatorial synthesis of highly selective cyclohexapeptides for separation of amino Acid enantiomers by capillary electrophoresis. AB - Cyclic peptide libraries dissolved in the electrolyte solution can be used as chiral selectors in capillary electrophoresis. In the present investigation, the resolution obtained in capillary electrophoresis for a set of dinitrophenyl-d,l amino acids was the parameter used to screen for the most effective selectors contained in a mixture of thousands of components of a cyclic hexapeptide sublibrary with three randomized positions. The deconvolution procedure was simplified by fractionating the sublibrary components according to the hydrophobicity of the amino acids in the randomized positions through reversed phase HPLC. By comparing the resolution obtained with the separated fractions, a set of hydrophobic amino acids was recognized as essential to achieve adequate enantioselectivity. The whole deconvolution process, which made it possible to select two highly selective cyclopeptides, required the synthesis and the evaluation of 15 sublibraries instead of the 54 syntheses required by a classical procedure of serial deconvolution. PMID- 21644679 TI - Rapid Prototyping of Microfluidic Systems in Poly(dimethylsiloxane). AB - This paper describes a procedure that makes it possible to design and fabricate (including sealing) microfluidic systems in an elastomeric material [Formula: see text] poly(dimethylsiloxane) (PDMS) [Formula: see text] in less than 24 h. A network of microfluidic channels (with width >20 MUm) is designed in a CAD program. This design is converted into a transparency by a high-resolution printer; this transparency is used as a mask in photolithography to create a master in positive relief photoresist. PDMS cast against the master yields a polymeric replica containing a network of channels. The surface of this replica, and that of a flat slab of PDMS, are oxidized in an oxygen plasma. These oxidized surfaces seal tightly and irreversibly when brought into conformal contact. Oxidized PDMS also seals irreversibly to other materials used in microfluidic systems, such as glass, silicon, silicon oxide, and oxidized polystyrene; a number of substrates for devices are, therefore, practical options. Oxidation of the PDMS has the additional advantage that it yields channels whose walls are negatively charged when in contact with neutral and basic aqueous solutions; these channels support electroosmotic pumping and can be filled easily with liquids with high surface energies (especially water). The performance of microfluidic systems prepared using this rapid prototyping technique has been evaluated by fabricating a miniaturized capillary electrophoresis system. Amino acids, charge ladders of positively and negatively charged proteins, and DNA fragments were separated in aqueous solutions with this system with resolution comparable to that obtained using fused silica capillaries. PMID- 21644680 TI - High-performance capillary gel electrochromatography with replaceable media. AB - Capillary gel electrochromatography is evaluated with an entangled polymer solution which is pumped into the capillary and run under fritless conditions. The polymer used has an acid backbone with grafted hydrophobic segments, the polyacid giving the electroosmotic flow and the hydrophobe segments providing the retentive component. Experimental evaluation of this type of system reveals performance similar to capillary electrophoresis and other forms of electrochromatography. The analysis of plate height data demonstrates that zone broadening is primarily due to diffusion with little contribution from nonequilibrium zone broadening. Hence, operation at high velocities (high voltages) is most desirable as opposed to most chromatographic methods. Some of the advantages of this type of experiment include being able to replace the retentive media in a few minutes, fast and reproducible high-performance separation, and having a retention mechanism similar to reversed-phase liquid chromatography. Disadvantages include a low retentive phase concentration and hence low sample loadability and limited solvent compatibility of the polymer. A number of different separations are demonstrated including separation of alkyl benzoates, alkylphenones, alkylbenzenes, oxidation inhibitors, and PAHs. PMID- 21644681 TI - Luminescent Mononuclear and Dinuclear Iridium(III) Cyclometalated Complexes Immobilized in a Polymeric Matrix as Solid-State Oxygen Sensors. AB - Oxygen quenching of the luminescence of mononuclear and dinuclear Ir(III) cyclometalated complexes immobilized in the pPEGMA matrixes has been studied. Linear Stern-Volmer plots, even when experiments at different emission wavelengths have been performed, were evidenced. Despite the different luminescence lifetimes of the chromophores in the absence of quencher, similar Stern-Volmer slopes have been calculated. This behavior was tentatively interpreted by taking into account the size and charge of the chromophores. Increased sizes and lower charges seem to enhance the sensitivity of the systems. Such findings could be of interest for the design of new solid-state luminescent oxygen sensors with improved performance. PMID- 21644682 TI - Microcalorimetry of chiral surfactant-cyclodextrin interactions. AB - The interactions of the chiral surfactants taurodeoxycholate (TDOCA) and deoxycholate (DOCA) with a range of cyclodextrins in aqueous solution have been investigated by isothermal titration microcalorimetry. In the presence of beta cyclodextrin, the apparent critical micelle concentration (cmc) of taurodeoxycholate is increased, and the enthalpy of demicellization decreased, in a manner consistent with 1:1 complexation of TDOCA with beta-CD at low concentrations. There is no evidence for direct interaction of cyclodextrins with surfactant micelles. This is confirmed by more direct binding titrations. Below the cmc, TDOCA forms 1:1 host-guest complexes with beta-cyclodextrin (DeltaH degrees (bind) = -32 kJ mol(-)(1), K(diss) = 0.38 mM; 25 degrees C, pH 7), methyl-beta-cyclodextrin (DeltaH(bind) = -13 kJ mol(-)(1), K(diss) = 0.36 mM), hydroxypropyl-beta-cyclodextrin (DeltaH degrees (bind) = -12 kJ mol(-)(1), K(diss) = 0.51 mM), and gamma-cyclodextrin (DeltaH degrees (bind) = -7.3 kJ mol( )(1), K(diss) = 0.08 mM), but not with the smaller alpha-cyclodextrin. At higher cyclodextrin concentrations, the calorimetric binding data are more ambiguous, suggesting 2:1 cyclodextrin/TDOCA complexation. Similar results are found with DOCA, though experiments here are limited by the tendency of DOCA to form gels in aqueous buffers. Enhanced chromatographic or electrophoretic chiral resolution observed in mixed chiral surfactant/cyclodextrin phases could be the result of increased solubility and/or the multiplicity of chiral complexes in such systems. PMID- 21644683 TI - Design and operation of a capsule-based microwave digestion system. AB - A high-pressure microwave digestion system has been designed based on the concept of using large-bore tubes as digestion vessels and capsules as a vehicle for sample introduction. Many of the design aspects are dictated by the use of a relatively large (8.4 mm o.d., 25 mm long) capsule, which in turn dictates the inner tube dimensions. A variety of materials were studied for use as the tubing material. PFA was selected as the best material for a demonstration arrangement. The cycle of operation involves insertion of the capsule by a flexible rod followed by addition of digestion reagent and then a heating/cooling/venting cycle for removal of gases. When the digestion is completed, the system removes the liquid with the same flexible rod and then cleans itself. The system is highly automated with computer-controlled venting, cooling, and reagent addition. Data indicate that performance is similar to that expected of a conventional microwave oven operated at the same temperatures. PMID- 21644684 TI - Individual detection of single-nanometer-sized particles in liquid by photothermal microscope. AB - We have developed a thermal lens microscope for liquid-phase and surface microanalyses. By applying the thermal lens microscope to particle detection, we succeeded in detecting a pulsed photothermal signal from single-nanometer-sized particles in liquid and counting them individually. The samples were polystyrene latex particles (190 and 80 nm in diameter) and colloidal Ag particles (10 nm in diameter). To verify that the detected pulsed signals corresponded to the single particle photothermal effects, we confirmed the items as described below using 190-nm polystyrene particles. First, no pulsed signal was generated under irradiation by either the excitation beam or the probe beam. Second, the pulse counts were proportional to the expectation value of the particles in the detection volume and zero for ultrapure water blank. Third, the pulse counts' distribution in a series of unit times had a Poisson distribution when the expectation value of the sample was much less than 1. Then, we demonstrated counting 80-nm polystyrene particles and 10-nm Ag particles in water. The pulsed signals were clearly distinguished from noise, and the signal-to-noise ratio was as large as 5. Finally, we discussed differences between the conventional thermal lens effect and the single-particle photothermal effect. Individual nanometer sized particle detection by photothermal effect was the first demonstration. PMID- 21644685 TI - Development of a dual-electrolysis stopped-flow method for the observation of electrogenerated chemiluminescence in energy-sufficient systems. AB - New methodology for the measurement of electrogenerated chemiluminescence (ECL) has been developed by using a dual-electrolysis stopped-flow system. Using this method, the ECL from "energy-sufficient" systems composed of different kinds of ion radicals can be easily observed by mixing both the electrolyzed solutions directly. The apparatus and method have been described in detail as well as the ECL observations for various energy-sufficient systems. In particular, in the reaction between the thianthrene (TH) cation radical and the pyrene (PY) anion radical, it was found that the emission spectra changed with the addition of the precursors, reflecting both complex electron- and energy-transfer processes in solution. The present results indicate that the electron- and energy-transfer reactions changed significantly, depending on the type of molecules. The ECL observation with the addition of the third molecule was also informative to compare the ease of the formation of the excited states. It was clarified that the excited states of 9,10-diphenylanthracene and rubrene are easily formed compared with those of TH and PY. PMID- 21644686 TI - Metal Speciation in the ppt Range by HPLC and Diode Laser Atomic Absorption Spectrometry in a Flame. AB - A simple, compact, and powerful instrument for metal speciation in the ppt range is described. The instrument includes a HPLC module for separation and a diode laser for element-selective detection by wavelength modulation absorption spectrometry in an analytical flame. The high detection power for metal species is due to a two-beam arrangement with logarithmic amplification of the normalized signal, which compensates the laser residual amplitude modulation noise, the offset, and its fluctuation. The analytical figures of merit are demonstrated by measurements of very low concentrations of Cr(VI) in tap water. PMID- 21644687 TI - Rugged gap reactor device for postcolumn fluorescence detection in capillary electrophoresis. AB - In this paper, the construction and performance of a rugged device for postcolumn derivatization in capillary electrophoresis (CE) are described. The device was based on a gap design, and a gap with a very small distance (<3 MUm, estimated under microscope) could be easily constructed without micromanipulation. Addition of derivatizing reagents into the reaction capillary was attributable to gravity flow. The concentration of derivatizing reagents can be controlled through manipulating the electroosmotic flow in the reaction capillary and the height of the liquid levels from the derivatizing reagents to the buffer reservoirs. The device has been applied in fluorescence detection of amino acids using a mixture of o-phthaldialdehyde/2-mercaptoethanol as derivatizing reagent. Theoretical plate numbers for 11 amino acids separated in a pH 9.5 borate buffer were obtained in the order of 40 000-250 000. The detection limit for glycine (S/N = 2) was found to be 6.7 * 10(-)(7) mol/L using a commercial HPLC fluorescence detector modified for CE. Free amino acids in a wine sample were also determined. Because the device is quite stable, we believe that it can be used routinely in analytical laboratories. PMID- 21644688 TI - Preparation and characterization of monolithic porous capillary columns loaded with chromatographic particles. AB - Using sol-gel technology, a porous glass matrix (xerogel) is formed in a capillary column and acts as a support for a stationary phase of chromatographic particles used in capillary electrochromatography. Preparation of the sol-gel matrix and immobilization of the octadecylsilica (ODS) stationary phase occur in a single step. The presence of the particles in the column greatly reduces matrix cracking caused by internal pressure differentials within the pores of the sol gel matrix. Good electroosmotic flow is achieved in part because of the inherent negative charge of both the particles and the sol-gel matrix. The performance of these sol-gel/ODS capillary columns was evaluated with a mixture of aromatic and nonaromatic organic compounds. Efficiencies of up to 80 000 plates/m were observed in columns with immobilized 3-MUm ODS particles. The efficiency and resolution are enhanced when 3-MUm ODS particles are used in place of the 5-MUm particles. PMID- 21644689 TI - Efficient computation of net analyte signal vector in inverse multivariate calibration models. AB - The net analyte signal vector has been defined by Lorber as the part of a mixture spectrum that is unique for the analyte of interest; i.e., it is orthogonal to the spectra of the interferences. It plays a key role in the development of multivariate analytical figures of merit. Applications have been reported that imply its utility for spectroscopic wavelength selection as well as calibration method comparison. Currently available methods for computing the net analyte signal vector in inverse multivariate calibration models are based on the evaluation of projection matrices. Due to the size of these matrices (p * p, with p the number of wavelengths) the computation may be highly memory- and time consuming. This paper shows that the net analyte signal vector can be obtained in a highly efficient manner by a suitable scaling of the regression vector. Computing the scaling factor only requires the evaluation of an inner product (p multiplications and additions). The mathematical form of the newly derived expression is discussed, and the generalization to multiway calibration models is briefly outlined. PMID- 21644690 TI - Protein Adsorption on Nanocrystalline TiO(2) Films: An Immobilization Strategy for Bioanalytical Devices. AB - We have investigated the use of optically transparent, nanoporous TiO(2) films as substrates for protein immobilization. Immobilization on such films may be readily achieved from aqueous solutions at 4 degrees C. The nanoporous structure of the film greatly enhances the active surface area available for protein binding (by a factor of 150 for a 4-MUm-thick film). We demonstrate that the redox state of immobilized cytochrome c may be modulated by the application of an electrical bias potential to the TiO(2) film and that the fluorescence yield of immobilized fluorophore-labeled maltose-binding protein may be used to monitor specifically maltose concentration. We conclude that nanoporous TiO(2) films may be useful both for basic studies of protein/electrode interactions and for the development of array-based bioanalytical devices employing both optical and electrochemical signal transduction methodologies. PMID- 21644691 TI - Fluorescence correlation spectroscopy for rapid multicomponent analysis in a capillary electrophoresis system. AB - We describe a new technique for performing multicomponent analysis using a combination of capillary electrophoresis (CE) and fluorescence correlation spectroscopy (FCS), which we refer to as CE/FCS. FCS is a highly sensitive and rapid optical technique that is often used to perform multicomponent analysis in static solutions based on the different diffusion times of the analyte species through the detection region of a tightly focused laser beam. In CE/FCS, transit times are measured for a mixture of analytes continuously flowing through a microcapillary in the presence of an electric field. Application of an electric field between the inlet and outlet of the capillary alters the transit times, depending on the magnitude and polarity of the applied field and the electrophoretic mobilities of the analytes. Multicomponent analysis is accomplished without the need to perform a chemical separation, due to the different electrophoretic mobilities of the analytes. This technique is particularly applicable to ultradilute solutions of analyte. We have used CE/FCS to analyze subnanomolar aqueous solutions containing mixtures of Rhodamine 6G (R6G) and R6G-labeled deoxycytosine triphosphate nucleotides. Under these conditions, fewer than two molecules were typically present in the detection region at a time. The relative concentrations of the analytes were determined with uncertainties of ~10%. Like diffusional FCS, this technique is highly sensitive and rapid. Concentration detection limits are below 10(-)(11) M, and analysis times are tens of seconds or less. However, CE/FCS does not require the diffusion coefficients of the analytes to be significantly different and can, therefore, be applied to multicomponent analysis of systems that would be difficult or impossible to study by diffusional FCS. PMID- 21644692 TI - Magnetic Sector ICPMS with Desolvating Micronebulization: Interference-Free Subpicogram Determination of Rare Earth Elements in Natural Samples. AB - A new method has been developed combining desolvating micronebulization with magnetic sector inductively coupled plasma mass spectrometry (ICPMS) for the analysis of all 14 stable rare earth elements (REEs) in small samples of marine particulate matter. Application is demonstrated for REEs in suspended particles from a deep ocean hydrothermal vent plume and a geological reference material. A 100-fold reduction in oxide formation, relative to standard nebulizer-spray chamber sample introduction, makes oxide interference correction negligible, even for samples that are very enriched in Ba and light REEs. Enriched isotopes for one light and one heavy REE ((145)Nd and (171)Yb) are used as both isotope dilution and internal standards, providing determination of all the REEs in one analysis. This standardization scheme eliminates the need for multimass drift correction used previously to achieve acceptable accuracy with external standardization techniques. Instead, the method exploits capabilities for accurate and precise determination of isotope ratios, a principal strength of ICPMS, and the mass-independent sensitivity of electric field scans on our double focusing instrument. We demonstrate overall precision of <=2% (1sigma) and accuracy better than 6% for all the REEs (except Er = 8.7%), based on comparison to recommended values for USGS certified reference material BHVO-1 (basalt). This performance is similar to that obtained by full isotope dilution mass spectrometric techniques, but the new method is far simpler, requires 5 min sample(-)(1), and avoids interferences introduced by complex mixtures of enriched isotopes. Sensitivity of (1.2-1.4) * 10(6) counts s(-)(1) ppb(-)(1) and background intensities of 2-60 counts s(-)(1) provide excellent detection limits of 1-40 ppq, a 100-fold improvement on established ICPMS methods. The low sample introduction rate (100 MUL min(-)(1)) allows unprecedented absolute detection limits of 1-20 fg. PMID- 21644693 TI - On-line nonmetal detection for argon supercritical fluid extraction using inductively coupled plasma optical emission spectroscopy. AB - This paper describes the development of a solventless instrumental method for determining organic contaminants in soil by coupling argon supercritical fluid extraction (SFE) to inductively coupled plasma atomic emission spectroscopy (ICP AES). In this method, organic compounds are first extracted by Ar SFE, transferred to the ICP directly in the supercritical (SC) argon, fragmented in the plasma, and then determined via their nonmetal atomic emissions. Supercritical Ar is superior to SC CO(2) for use in ICP-AES because it overcomes the disadvantages of plasma blowout, noisy baselines, and CO(2) interference in the determination of carbon. All instruments employed were commercially available, and the interface between the SFE and the torch of the ICP consisted of a simple glass capillary tube. Four nonmetals, C, S, P, and Si, were selected for this preliminary study. The selectivities obtained for these nonmetals, referenced to carbon, were found to be 345, 38 000, and 1400 for sulfur, phosphorus, and silicon, respectively. With the exception of carbon, the mass detection limits are in the range of 0.06-1.8 MUg. For this work, the mass detection limit of carbon is 66 MUg and is limited by the smallest aliquot of material that can be injected by syringe. Finally, total petroleum hydrocarbons (TPHs) were determined in two "real world" contaminated soil samples via the carbon emission at 247.9 nm. Dodecane was used as the standard reference compound because its response was found to be similar to other petroleum hydrocarbons (e.g., kerosene, gasoline, no. 1 fuel oil, and no. 2 fuel oil). Additionally, it was found necessary to add a small amount (i.e., 250 mg) of drying agent to the SFE vessel in order to prevent the plasma from extinguishing from the extracted water. The results of the on-line Ar SFE/ICP-AES determinations of TPH in the two samples agreed well with those obtained from conventional off-line CO(2) SFE and off-line Ar SFE. PMID- 21644694 TI - Computer simulations of electrokinetic transport in microfabricated channel structures. AB - A mathematical model describing electrokinetically driven mass transport phenomena in microfabricated chip devices is presented in this paper. The model accounts for principal material transport mechanisms such as electrokinetic migration (electrophoresis and electroosmosis) and diffusion. A computer code that implements the model is capable of simulating transport of materials during electrokinetic manipulation in 2-D channel structures. The computer code allows arbitrary channel geometries with various boundary conditions for the electric field and the sample concentration. Two fundamental microfluidic chip elements, a cross and a mixing tee, are of particular interest. An electrokinetic focusing experiment using a cross structure and mixing in a tee structure are simulated. Simulations revealed an optimum focusing voltage for which the ratio of sample concentration to sample width is maximized. They also verified that the mixing tee provides very accurate dilution/mixing characteristics for both charged and neutral samples. Good agreement between simulated and experimental data verified the accuracy of the mathematical model. PMID- 21644695 TI - Fluorometric detection of ca(2+) based on an induced change in the conformation of sol-gel entrapped parvalbumin. AB - We report the development of a fluorometric detection strategy for Ca(2+) based on induced changes in the conformation of cod III parvalbumin entrapped within a sol-gel processed glass. The detection scheme utilizes a fluorescent allosteric signal transduction (FAST) strategy wherein conformational changes induced by Ca(2+) binding result in alterations in the intrinsic fluorescence from the single tryptophan residue at position 102. Intrinsic fluorescence was also used to examine chemically induced changes in protein structure to ascertain the effects of entrapment on the conformational motions and stability of the protein. Fluorescence analysis indicated that the behavior of the protein depended on the entrapment protocols used. The entrapped protein retained conformational flexibility similar to that observed in solution and remained accessible to analytes such as Ca(2+). Entrapment also caused improvements in protein stability against chemical denaturants. However, entrapment caused the apparent affinity constant for binding of Ca(2+) to decrease substantially with aging time. Even so, in optimum cases, fluorometric detection of Ca(2+) could be done over a 600 MUM range with a limit of detection of 3 MUM and with no interference from divalent ions such as Mg(2+), Sr(2+), or Cd(2+), indicating the viability of using sol-gel entrapped FAST proteins for the detection of Ca(2+). PMID- 21644696 TI - Analysis of Xanthate Derivatives by Vacuum Ultraviolet Laser-Time-of-Flight Mass Spectrometry. AB - A series of liquid O,S-dialkyl dithiocarbonates (xanthate esters) have been synthesized, and time-of-flight (TOF) mass spectra were recorded for their vapors using both ultraviolet (UV) and vacuum ultraviolet (VUV) laser excitation. These compounds are chemical derivatives of low vapor pressure xanthate salts which have found important commercial application as collectors in mineral sulfide flotation circuits. Our experiments demonstrate that esters ionized by short wavelength VUV light can be detected by parent mass with high efficiency and minimal fragmentation. In contrast, the mass spectra of the same compounds obtained by UV light excitation exhibit a large number of low molecular mass peaks. A preliminary quantitative analysis of the composition of a gas-phase mixture of xanthate esters has also been achieved, which indicates possible subfemtomole detection limits. PMID- 21644697 TI - Simultaneous separation of acidic, basic, and neutral organic compounds, including strong and moderate acids and bases, by capillary electrochromatography. AB - The separation of strongly basic, moderately basic, weakly basic, strongly acidic, moderately acidic, weakly acidic, and neutral compounds in a single run using capillary electrochromatography (CEC) is presented. This is accomplished using a 3-MUm CEC Hypersil C8 capillary with high organic content acetonitrile/phosphate (pH 2.5) mobile phases containing hexylamine. Fifteen basic, acidic, and neutral drugs of forensic interest are resolved using a step gradient. Strong and moderately basic drugs separate before t(o), apparently by a combination of free zone electrophoresis (CZE) and chromatographic phenomena. Weak bases separate after t(o), also by a combination of CZE and chromatographic processes. Due to large selectivity differences between CEC and CZE for bases, there is evidence that the stationary phase is playing a significant role in the separation of these solutes. The CEC approach presented offers unique selectivity, expanded peak capacity, and the ability to solubilize both hydrophilic and hydrophobic solutes in an injection solvent that is compatible with the chromatographic system. PMID- 21644698 TI - Summarizing the effectiveness of supercritical fluid extraction of polycyclic aromatic hydrocarbons from natural matrix environmental samples. AB - A summary of the supercritical fluid extraction (SFE) of polycyclic aromatic hydrocarbons (PAHs) from four natural matrix Standard Reference Materials (SRMs) is presented. The work involved the investigation of the effects of extraction fluid [carbon dioxide (CO(2)), chlorodifluoromethane (R22), and 1,1,1,2 tetrafluoroethane (HFC134a)], fluid modifier (dichloromethane and aniline), temperature (60, 150, and 200 degrees C) and added water on the SFE recoveries of PAHs compared to certified results from Soxhlet extractions. For SRM 1649a (Urban Dust/Organics), R22 yielded excellent recoveries (>90% of certified concentrations) of all PAHs measured, while results for the same SRM using HFC134a as the fluid were typically <80% of the certified concentrations for most of the PAHs measured. For SRM 1941a and 1944, both aquatic sediments with similar physical and chemical compositions, extractions of the wet materials with dichloromethane-modified CO(2) (10%, v/v) yielded quantitative recoveries of all PAHs for SRM 1944 but an obvious trend of lower recoveries for higher molecular weight PAHs (>=228 amu) for SRM 1941a. Results of SFEs of SRM 1650 (Diesel Particulate Matter) showed that this material is the most refractory of the SRMs investigated in this study, with recoveries of indeno[1,2,3-cd]pyrene and benzo[ghi]perylene at <20% of the Soxhlet results. PMID- 21644699 TI - Discrimination between Butylammonium Isomers by Calix[5]arene-Based ISEs. AB - Penta-O-alkylated p-tert-butylcalix[5]arenes 1-5 (R = benzyl, isohexyl, isopropoxyethyl, isopropoxycarbonylmethyl, and tert-butoxycarbonylmethyl, respectively) in a fixed C(5)(v) cone conformation have been studied as ionophores in liquid membrane ion-selective electrodes (ISEs) for n-butylammonium against the other branched butylammonium isomers, alkali metals, and ammonium ions, in terms of detection limits, sensitivity, and selectivity. The highest levels of potentiometric selectivity and detection limits up to 3 * 10(-)(6) M are observed with ISEs based on ionophore 2, where selectivity follows the order n-BuNH(3)(+) ? i-BuNH(3)(+) > s-BuNH(3)(+) > t-BuNH(3)(+). The lower potentiometric selectivity displayed by ISEs based on ionophores 3-5 is ascribed to their affinity for the Na(+) ion of the lipophilic salt present in the membrane, as evidenced by appropriate (1)H NMR competition experiments with Na(+) and n-BuNH(3)(+) ions. Further investigation on the selectivity mechanism of ionophore 2 by means of frequency response analysis shows that the interaction of the linear butylammonium ion with membranes containing 2 involves a lower resistance process than that occurring with the other branched isomers, thus suggesting the presence of a favorable kinetic-controlled mechanism. PMID- 21644700 TI - Resonant cavity gas-phase polarimeter. AB - A high-sensitivity polarimeter is demonstrated for application to gas-phase chirality measurement. This device is based on the physics of the eigenstates of a Fabry-Perot cavity, permitting improvement in the sensitivity with respect to the usual polarimeters. Typical measurements of rotations of 50 (+/-1) * 10(-)(5) degrees induced by the optical activity of (R)-(+)-limonene and (S)-(-)-limonene in the vapor phase are shown. A noise level corresponding to a rotation of 10( )(6) degrees is experimentally demonstrated. Application to the polarimetric monitoring of an enantiomer mixing racemization of limonene in the gas phase is also presented. PMID- 21644701 TI - Method for the elimination of chromatographic bias from measured capillary electrophoretic effective mobility values. AB - A method based on a modified version of pressure-mediated capillary electrophoresis (PreMCE) has been developed for the elimination of the chromatographic bias inherent in effective electrophoretic mobilities measured by capillary electrophoresis. This new five-band PreMCE method can be readily executed on most commercial capillary electrophoresis instruments. It yields not only precise but also accurate effective mobilities and electroosmotic flow rates, even when the analytes and electroosmotic flow markers are strongly retained on the coated fused-silica capillary wall. PMID- 21644702 TI - Fourier transform detection in a cylindrical quadrupole ion trap. AB - Broad-band nondestructive ion detection based on induced image current measurement is performed in a quadrupole ion trap having cylindrical geometry. Spectra of krypton and acetophenone are shown to demonstrate the first use of nondestructive detection with a cylindrical ion trap. PMID- 21644703 TI - A new method of determining chlorine kinetic isotope effects. AB - Two methods have been used to measure the chlorine leaving group kinetic isotope effect for the S(N)2 reduction of benzyl chloride to toluene by sodium borohydride in DMSO at 30.000 degrees C. The reaction was monitored by titrating the unreacted borohydride ion. One method involved determining the chlorine isotope effect using the classical IRMS method, which requires the conversion of the chloride ions into gaseous methyl chloride that is analyzed in an isotope ratio mass spectrometric analyses (Hill, J. W.; Fry, A. J. Am. Chem. Soc. 1962, 84, 2763. Taylor, J. W.; Grimsrud, E. P. Anal. Chem. 1969, 41, 805.). Two different measurements using this method yielded isotope effects of k(35)/k(37) = 1.007 19 +/- 0.000 19 and 1.007 64 +/- 0.000 19. The second method was a new technique where the ratio of the chlorine isotopes was obtained by fast atom bombardment mass spectrometry on the silver chloride recovered from the reaction, i.e., from the first step in the classical procedure. Therefore, the new method is much simpler and avoids the time-consuming preparation, purification, and recovery of the gaseous methyl chloride. Although the experimental error is larger (k(35)/k(37) = 1.008 03 +/- 0.00 10 and 1.008 02 +/- 0.000 65) when the new technique is used to analyze the silver chloride samples from the same set of experiments that were used to measure the isotope effect by the classical method, the chlorine isotope effect found by the two methods is identical within experimental error. This large chlorine kinetic isotope effect indicates there is considerable C(alpha)-Cl bond rupture in the S(N)2 transition state. PMID- 21644704 TI - The use of multidimensional liquid-phase separations and mass spectrometry for the detailed characterization of posttranslational modifications in glycoproteins. AB - The goal of characterization of the proteome, while challenging in itself, is further complicated by the microheterogeneity introduced by posttranslational modifications such as glycosylation. A combination of liquid chromatography (LC), capillary electrophoresis (CE), and mass spectrometry (MS) offers the advantages of unique selectivity and high efficiency of the separation methods combined with the mass specificity and sensitivity of MS. In the current work, the combination of liquid-phase separations and mass spectrometry is demonstrated through the on line coupling of electrospray ionization mass spectrometry (ESI-MS) and off-line coupling with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF-MS). LC/ESI-MS yields real-time results while maintaining the separation obtained from the LC analysis. CE/MALDI TOF-MS offers high-mass detection and extremely low detection limits. The unique separation selectivity of CE relative to reversed-phase HPLC separations of the members of a glycopeptide family was used to develop an integrated multidimensional analysis achieved by the off-line coupling of LC, CE, and MALDI TOF-MS. To demonstrate the applicability of these techniques to the characterization of the heterogeneity of posttranslational modifications present in glycoproteins, we will report on the study of the glycoforms present in a N-linked site in a single-chain plasminogen activator (DSPAalpha1). PMID- 21644705 TI - Capillary electrophoretic determination of different classes of organic ions by potentiometric detection with coated-wire ion-selective electrodes. AB - Singly charged amines and sulfonic acids as cationic and anionic aliphatic model substances, respectively, were detected in capillary electrophoresis with all solid-state ion-selective electrodes. The sensitivity for these compounds is a function of their lipophilicity. The signal detected is generally greater for molecules with a larger organic part. The utility of the method is further demonstrated by the detection of a group of aromatic compounds in the form of the anionic analgesics (S)-(+)-2-(4-isobutylphenyl)propionic acid, 2-[(2,6 dichlorophenyl)amino]benzeneacetic acid, and o-acetylsalicylic acid. Further determined were the artificial sweeteners cyclamate, acesulfame-K, and saccharin. Detection limits for the different substances were between 2.5 * 10(-)(6) and 1 * 10(-)(5) M. PMID- 21644706 TI - Complexation between Amylodextrin Oligomers and Selected Pharmaceuticals Measured through Capillary Electrophoresis. AB - The amylodextrin oligomers, tagged fluorescently at their reducing ends for laser fluorescence detection, were used as model solutes to investigate certain carbohydrate-drug interactions through capillary electrophoresis. The separation patterns of some oligomers were found to be altered significantly by the presence of selected pharmaceuticals used as buffer additives. The selectivity of complexation at a particular size of an oligomer could be judged from alterations in migration times and peak shapes. The formation of complex was influenced substantially by solutes' chemical environment (pH, ionic strength, and the nature and concentration of organic additives). The chemical nature of the guest molecules also played an important role in complexation. Using amylodextrins as chiral selectors, enantiomeric resolution of several pharmaceuticals was achieved electrophoretically. The conclusions drawn from electrophoretic data were found to be in close agreement with the results of a (13)C NMR study. PMID- 21644707 TI - Enantiomeric separation of sulfonium ions by capillary electrophoresis using neutral and charged cyclodextrins. AB - Capillary zone electrophoresis was successfully applied, for the first time, to the chiral separation of structurally related sulfonium ions, using sodium phosphate buffer pH 2.5 with beta-cyclodextrin (beta-CD) or sulfated-beta cyclodextrin (S-beta-CD) as the chiral selector with tetrabutylammonium bromide (TBA). For this study, a series of structurally related sulfonium ions in which one of the alkyl chains varied in length were synthesized from a common sulfide. The resolution of the ions was found to be dependent on the type of cyclodextrin used, the presence or absence of TBA, and the structure of the sulfonium ion. beta-CD was found to be effective only for ions containing two aromatic groups, while the S-beta-CD was effective only for ions containing one aromatic group. The chiral separation of thiophenium ions was also studied under the conditions established. Chiral separation of sulfonium ions in a binary buffer system containing methanol was explored as well. Separations were achieved for all but one sulfonium ion and one thiophenium ion. The data presented show the effectiveness of the enantiomeric separation using CZE with cyclodextrin-modified buffer for these types of ions. PMID- 21644708 TI - Electrically polarized ion-exchange beds in ion chromatography: eluent generation and recycling. AB - This paper describes how electrically polarized ion-exchange beds pumped with water can produce electrolyte of steady and controllable concentration. Such devices make it possible to use water as the pumped phase in ion chromatography (IC), thus avoiding off-line eluent preparation. Control of the electrical current flowing through the devices allows precise control of the concentration of eluent that they deliver. This provides a new way of performing gradient and isocratic elutions. Using water as the carrier and two small beds of resin, one as a generator the other as a suppressor, and periodically reversing their roles through automatically switched valves, we have developed a form of continuous IC that involves little intervention by the user. The paper presents the principles of the new method and examples of its use in anion analysis. PMID- 21644709 TI - Determination of theophylline in serum by molecularly imprinted solid-phase extraction with pulsed elution. AB - The technique of molecular imprinting is used to produce an extensively cross linked poly(methacrylic acid-co-ethylene dimethacrylate) material that contains theophylline as a print molecule. After Soxhlet extraction of the theophylline, binding sites are formed in the polymer with complementary size, shape, and positioning of chemical functionalities. The molecularly imprinted polymer's (MIP) high theophylline selectivity, chemical stability, and physically robust nature make it an ideal stationary-phase material in columns for HPLC separation of theophylline from other structurally related drug compounds. Mobile-phase tests confirm that a retention mechanism typical of normal-phase chromatography governs the separation, and selectivity of the MIP column can be controlled by a combination of the mobile phase and the sample solvent. Under optimal conditions, the MIP column functions like a solid-phase sorbent for theophylline extraction. Rapid elution of the bound theophylline can be accomplished in a pulsed format through injection of 20 MUL of a solvent that has the proper polarity and protic nature to disrupt the electrostatic interactions and hydrogen bonding between theophylline and binding sites. A concentration detection limit of 120 ng/mL is obtained using direct UV absorption detection at 270 nm, which corresponds to a mass detection limit of 2.4 ng. This new technique, molecularly imprinted solid phase extraction with pulsed elution (MISPE-PE), permits on-line preconcentration of theophylline from a large volume of dilute sample solution. Using a sample volume of 300 MUL, a 40 ng/mL standard solution produces a detectable peak signal. Application of MISPE-PE in serum analysis further demonstrates the high capability of the MIP column to selectively isolate theophylline from other matrix components for fast, accurate determination. PMID- 21644710 TI - Polymer Degradation Mechanism Reselection through Derivatization for Qualitative Pyrolysis Gas Chromatography Analysis. AB - A poly(vinylcarboxylic acid) and its analogue alkyl ester exhibited significantly different thermal degradation mechanisms under pyrolysis conditions. The greatly enhanced monomer production of poly(alkylvinylcarboxylate) during pyrolysis makes possible qualitative analysis of poly(vinylcarboxylic acid) through its derivatized alkyl ester. In this study, copolymers containing poly(methacrylic acid) and methacrylic acid were used to demonstrate the pyrolysis gas chromatography qualitative analysis of derivatized polymer. The thermal decomposition mechanisms of poly(methacrylic acid), and poly(methyl methacrylate) were studied. The advantages and disadvantages of this derivatized process as related to Py-GC analysis are discussed. PMID- 21644711 TI - Determination of Anionic, Neutral, and Cationic Species of Arsenic by Ion Chromatography with ICPMS Detection in Environmental Samples. AB - Ion chromatographic methods developed to separate either cationic, neutral, and anionic arsenic species or soluble and suspended arsenic species were successfully used in DORM-2 standard reference material and in water samples of environmental interest. The most effective separation of the analytes within 10 min was achieved with a nitric acid gradient elution using a strong anion exchange stationary phase with additional capacity for hydrophobic interactions (IonPac AS7). The elemental-specific detection mode allows the sensitive determination of the arsenic species in the submicrogram per liter range. The calibration results were compared with those obtained by an alkaline water methanol mixed eluent combined with a weak anion-exchange column (IonPac AS4A SC). Differences in sensitivities were eclipsed by the low level of the baseline and the noise when using nitric acid. The gradient method was used to determine arsenic species in highly ferrous/ferric-contaminated leachates of lignite spoil. The companion elements underwent parallel screening to explain the interactions of arsenic species with the major elements. PMID- 21644712 TI - Measurement of diffusive flux of ammonia from water. AB - An instrument was developed for the measurement of gaseous ammonia concentration, NH(3(sw,eq)), in equilibrium with surface waters, notably ocean water. The instrument measures the ammonia flux from a flowing water surface under defined conditions and allows the calculation of NH(3(sw,eq)) from the principles of Fickian diffusion. The flux collector resembles a wetted parallel plate denuder previously developed for air sampling. The sample under study runs on one plate of the device; the ammonia released from the sample is collected by a slow flow of a receptor liquid on the other plate. The NH(3) + NH(4)(+) (hereinafter called N(T)) in the effluent receptor liquid is preconcentrated on a silica gel column and subsequently measured by a fluorometric flow injection analysis (FIA) system. With a 6-min cycle (4-min load, 2-min inject), the analytical system can measure down to 0.3 nM N(T) in the receptor liquid. Coupled with the flux collector, it is sufficiently sensitive to measure the ammonia flux from seawater. The instrument design is such that it is little affected by ambient ammonia. In both laboratory (N(T) 0.2-50 MUM), and field investigations (N(T) 0.18-1.7 MUM) good linearity between the ammonia flux and the N(T) concentration in seawater (spiked, synthetic, natural) was observed, although aged seawater, with depleted N(T) content, behaves in an unusual fashion upon N(T) addition, showing the existence of an "ammonia demand". NH(3(sw,eq)) levels from ocean water measured in the Coconut Island Laboratory, HI, ranged from 6.6 to 33 nmol/m(3) with an average of 17.4 +/- 6.9 nmol/m(3), in comparison to 2.8-21 nmol/m(3) (average 10 +/- 7 nmol/m(3)) NH(3(sw,eq)) values previously reported for the Central Pacific Ocean (Quinn, P. K.; et al. J. Geophys. Res. 1990, 95, 16405-16416). PMID- 21644713 TI - Methods for determining the intrinsic and effective charges on spherical macroions. AB - The intrinsic number of charges per particle, Z, on particles in a suspension of monodisperse sulfonated polystyrene latex is found by measuring steady-state voltammetric transport-limited currents for the reduction of hydrogen counterion at a Pt disk microelectrode in a suspension containing excess supporting electrolyte. Limiting currents measured in deionized latex suspensions yield a corresponding effective charge, Z*. Electrostatic binding of an inner layer of counterions to the particle renders Z* < Z. Voltammetrically determined charges agree with the intrinsic and effective Stokes charges as determined by titration and electrophoresis, respectively. zeta- Potentials calculated from the measured electrophoretic mobility of the particles yield the Loeb charge number, which agrees more closely with Z than with Z*. For existing data on spherically charged macroions, Z*/Z decreases with increasing ratio of intrinsic charge to macroion radius, Z/a. This finding is supported by effective charge values calculated from the cell model using the nonlinear Poisson-Boltzmann equation with the convention that Z* is the charge found between the c(r)/ = 1 boundary and the cell wall. PMID- 21644714 TI - Capacitive Approach To Determine Phospholipase A(2) Activity toward Artificial and Natural Substrates. AB - A capacitive approach has been employed to develop a novel method to determine phospholipase activity. The sensing electrodes have a structure like Au/S(CH(2))(17)CH(3)/substrate/electrolyte. Hydrolysis of the substrate, mediated by phospholipase A(2), leads to the formation of water-soluble products from the insoluble substrate. This results in desorption of these products into aqueous phase and corresponding increase of the electrode capacitance. The requirement of high water solubility of the reaction products can be achieved in two ways. In the first, short-chain phospholipids are used as the substrate, in which case, water-soluble products are formed and no additional reagents are required to promote desorption of these products. The sensors prepared by this strategy provide sensitive qualitative detection of phospholipases. The second way is based on the use of a water-soluble acceptor (for example, beta-cyclodextrin) to solubilize the products of hydrolysis. It allows semiquantitative detection of phospholipase activity toward long-chain natural substrates. The reaction kinetics for this case was found to be monoexponential and linearly dependent on the phospholipase concentration. The detection limit of this method, as tested with phospholipase A(2) from bee venom and soy bean lecithin as the substrate, is ~0.5 ng/mL (500 MUunits/mL). PMID- 21644715 TI - Continuous-flow/stopped-flow and rotating bioreactors in the determination of glucose. AB - The high sensitivity that can be attained by enzymatic amplification via substrate cycling has been verified by on-line interfacing of a rotating bioreactor and continuous-flow/stopped-flow/continuous-flow processing [Raba, J.; Mottola, H. A. Anal. Biochem. 1994, 220, 297-302]. The determination of glucose levels was possible with a limit of detection of 0.2 fmol.L(-)(1) in the processing of as many as 30 samples per hour. Determination at such low levels is of interest in several situations encountered in fermentation biotechnology and clinical chemistry, and this determination in culture broths illustrates the capabilities of the proposed approach. The glucose oxidase/glucose dehydrogenase coupled system was used by immobilizing glucose oxidase (EC 1.1.3.4) on the top of a rotating disk while glucose dehydrogenase (EC 1.1.1.47) was immobilized on the top part of the flow-through cell. Substrate cycling was realized via NADH/NAD(+) that, in conjunction with glucose dehydrogenase, regenerates glucose, the substrate in the glucose oxidase-catalyzed reaction. This cycling permits generation of H(2)O(2) (detected at Pt ring electrode concentric to the rotating disk) beyond stoichiometric limitations. This permits a 100-fold increase in the sensitivity for glucose determination when compared with the determination involving no substrate cycling. PMID- 21644716 TI - An Enzyme Switch Employing Direct Electrochemical Communication between Horseradish Peroxidase and a Poly(aniline) Film. AB - An enzyme switch, or microelectrochemical enzyme transistor, responsive to hydrogen peroxide was made by connecting two carbon band electrodes (~10 MUm wide, 4.5 mm long separated by a 20-MUm gap) with an anodically grown film of poly(aniline). Horseradish peroxidase (EC 1.11.1.7) was either adsorbed onto the poly(aniline) film or immobilized in an insulating poly(1,2-diaminobenzene) polymer grown electrochemically on top of the poly(aniline) film to complete the device. In the completed device, the conductivity of the poly(aniline) film changes from conducting (between - 0.05 and + 0.3 V vs SCE at pH 5) to insulating (>+0.3 V vs SCE at pH 5) on addition of hydrogen peroxide. The change in conductivity is brought about by oxidation of the poly(aniline) film by direct electrochemical communication between the enzyme and the conducting polymer. This was confirmed by measuring the potential of the poly(aniline) film during switching of the conductivity in the presence of hydrogen peroxide. The devices can be reused by rereducing the poly(aniline) electrochemically to a potential below +0.3 V vs SCE. A blind test showed that the device can be used to determine unknown concentrations of H(2)O(2) in solution and that, when used with hydrogen peroxide concentrations below 0.5 mmol dm(-)(3), the same device maybe reused several times. The possible development of devices of this type for use in applications requiring the measurement of low levels of hydrogen peroxide or horseradish peroxidase is discussed. PMID- 21644717 TI - Stacking from the Sample Stream in CZE Using a Pneumatically Driven Computerized Sampler. AB - It is demonstrated that a pneumatically driven computerized sampling device for capillary electrophoresis facilitates sample stacking by the head-column field amplification (HCFA) technique. This device utilizes a rapid exchange between buffer and sample in a narrow channel at the separation capillary inlet and makes possible the combination of two classical injection modes [Formula: see text] the electrokinetic and hydrodynamic modes. Detection limits obtained were about 9 nM for alkylbenzylamine cations with common UV detection. PMID- 21644718 TI - Complete elimination of interferences in the organotin determination by oxidation with dimethyldioxirane combined with alumina cleanup. AB - Most of the analytical procedures used in organotin (OT) speciation from sediment involves the Grignard derivatization reaction followed by a cleanup step and a desulfuration reaction since sulfur and/or sulfur species interfere with OT determination by GC/MS or GC-FPD. However, alkyl sulfides are generated from the coextracted elemental sulfur, and they are not removed by conventional desulfurization procedures. We propose here a method based on the oxidation of all the sulfur species with dimethyldioxirane (DMD) to sulfones or sulfur oxides. While sulfones are easily eliminated by alumina adsorption chromatography because they have higher polarity than OTs, the sulfur oxides are spontaneously evaporated. The DMD chemoselectivity favors the oxidation of sulfur compounds to sulfones in a few minutes, whereas OTs remain unreacted. In addition, the excess DMD is easily removed by evaporation under a nitrogen stream before the Al(2)O(3) cleanup step. The effectiveness of the desulfurization reaction combined with the cleanup step is demonstrated for a variety of sediment samples containing up to 3.1% of elemental sulfur, which is completely removed by adding 0.6 molar equiv of DMD. No statistical differences in the OT distribution pattern throughout the DMD intermediate oxidation steps were observed. PMID- 21644719 TI - Selective removal of cesium from Acid solutions with immobilized copper ferrocyanide. AB - Copper ferrocyanide can be immobilized on Chelex-20, a technical grade chelating resin, for selective removal of cesium from neutral to acidic solutions. The immobilized copper ferrocyanide is much easier to handle than the copper ferrocyanide powder in extraction and separation of cesium from aqueous solutions. In the presence of a reducing agent such as hydrazine, the immobilized copper ferrocyanide resin is able to remove 98% of the cesium from a simulated acid waste solution in one cycle of batch experiments. The resin shows promising properties for remediation of acidic nuclear wastes and for concentration of (137)Cs from environmental samples for quantitative analysis. PMID- 21644720 TI - A comparison of gas-hexadecane and gas-apolane partition coefficients. AB - Gas-apolane partition coefficients (L(87)) of 157 nonpolar and polar organic solutes, spanning a wide range of functional groups, dipolarities, and hydrogen bonding capabilities, are measured by open tubular capillary gas chromatography at 40 degrees C. The experimental values compare well (R(2) = 0.999) with literature values of log L(87) from packed-column gas chromatography. The log L(87) values are also compared with gas-hexadecane partition coefficients (log L(16)) at 25 degrees C. A strong linear relationship exists between log L(87) and log L(16) for all solutes (R(2) = 0.994), as well as for chemically relevant subsets of the data. Therefore, unknown L(16) values at 25 degrees C can be predicted from the corresponding L(87) values, which can be measured on open tubular or packed columns at higher temperatures, due to the low volatility of apolane. Predicted values of L(16) would be extremely useful, since log L(16) is often the major explanatory parameter in many linear solvation energy relationships. PMID- 21644721 TI - Molecular Design, Characterization, and Application of Multiinformation Dyes for Multidimensional Optical Chemical Sensings. 2. Preparation of the Optical Sensing Membranes for the Simultaneous Measurements of pH and Water Content in Organic Media. AB - Optical chemical sensing of pH and water content in organic solvents is proposed, using multiinformation dyes (MIDs) based on the support matrixes for the dyes. In this investigation, four kinds of merocyanine-type dyes having a polymerizable olefin unit as the MIDs were synthesized. These dyes were copolymerized with hydrophilic monomer molecules to obtain dye-immobilized optical chemical sensor (optode) membranes. In this case, selection of the monomer molecule gave optode membranes having different color change properties, because different monomer molecules provided different chemical environments around the immobilized dye. These optode membranes were used for the measurement of pH and water content in organic solvents. These membranes offered two-dimensional sensing information in one spectrum when they were employed for water content sensing in organic solvents, in which the maximum wavelength represents the water content and the absorbance at this wavelength represents the pH of the water present. These polymer membranes have a long lifetime, which can be adequate for practical use. PMID- 21644722 TI - Correlation between Extraction Equilibrium of Uranium(VI) and Density of CO(2) Medium in a HNO(3)/Supercritical CO(2)-Tributyl Phosphate System. AB - The extraction equilibrium of U(VI) between a nitric acid solution and a supercritical CO(2) phase containing tributyl phosphate (TBP) is formulated taking into account that (i) a distribution ratio of a metal extracted is a function of a distribution constant of each component involved in the extraction reaction, (ii) the distribution constant is defined as a ratio of solubilities of the component in both phases, and (iii) the solubility in the CO(2) phase is correlated with density of CO(2). A simple linear relationship between the distribution ratio, D(U), of U(VI) and density, rho, of CO(2) is derived; log D(U) = a log rho + A + B, in which a is a proportional constant implying the solvation characteristic of the solute in supercritical CO(2), A is a pressure independent constant, and B is a variable determined by the distribution equilibrium of HNO(3). The equation derived was verified experimentally by the measurement of the distribution ratio of U(VI) and HNO(3) under various conditions of pressure and temperature. A novel concept of selective supercritical fluid extraction of metals by means of pressure-tuning or CO(2) density-tuning was proposed. PMID- 21644723 TI - Highly Sorbent Films Derived from Ni(SCN)(2)(4-picoline)(4) for the Detection of Chlorinated and Aromatic Hydrocarbons with Quartz Crystal Microbalance Sensors. AB - A quartz crystal microbalance (QCM) spray-coated with a Ni(SCN)(2)(4-picoline)(4) film is a sensitive detector for small aromatic (benzene, toluene) and chlorinated (trichloroethylene, perchloroethylene) vapors with a planar molecular geometry. Frequency changes during transient exposures to these vapors are rapid and reversible. In contrast, frequency changes during transient exposures to carbon tetrachloride vapor exhibit a very slow rise and decay. Impedance studies demonstrate that the QCMs are responding only to mass changes in the film. Calibration curves exhibit both linear and near-saturation responses, depending on the vapor and vapor concentration. Partition coefficients obtained from the linear response regimes of the calibration curves are in the 10 000-100 000 range, more than an order of magnitude larger than the partition coefficients for a prototypical soft polymer, poly(isobutylene). Despite the absence of evidence for crystallinity by optical or X-ray diffraction methods, the spray-coated films appear to be forming clathrates with the organic vapors. The Ni(SCN)(2)(4 picoline)(4) film is promising for the development of very sensitive and partially selective piezoelectric sensors for nonpolar or weakly polar organic vapors in air. PMID- 21644724 TI - Validation of analytical methods using a regression procedure. AB - The evaluation and validation of analytical methods and instruments require comparison studies using sample material for testing accuracy and precision. In analytical chemistry, the commonly accepted means of analyzing data from method comparison studies is least-squares regression analysis, a model which has limitations. In this paper, the results from ordinary least-squares and many other regression approaches recommended in the literature were compared with a new regression procedure that takes into account the errors in both variables (methods). After a discussion of the properties of the regression procedure, recommendations are given for carrying out a method comparison study using informational analysis of variance. The efficiency of the regression procedure proposed is demonstrated by applying it to different data sets from published literature. PMID- 21644725 TI - Fluorescence correlation spectroscopy with patterned photoexcitation for measuring solution diffusion coefficients of robust fluorophores. AB - Patterned fluorescence correlation spectroscopy is developed as a new technique for measuring diffusion coefficients of photostable fluorescent probe molecules. In this method, interference between two intersecting, coherent laser beams creates an excitation fringe pattern from which fluorescence emission is monitored. Spontaneous concentration fluctuations of fluorescent molecules within the excitation volume are detected as excess noise on a fluorescence transient; concentration fluctuations are driven primarily by diffusion of these molecules between interference fringes although contributions from photobleaching and diffusion over the entire pattern dimensions can also be observed. Autocorrelation of the fluorescence transient allows analysis of the temporal characteristics of the fluctuations, which were used to determine solution diffusion coefficients; the method was applied to study the diffusion of Rhodamine 6G (R6G) in water/methanol solutions containing added electrolyte and in pure ethanol. The method can be used to characterize the diffusive transport of fluorescently labeled species, which is an important issue in designing small volume detection experiments. PMID- 21644726 TI - Temperature-constrained cascade correlation networks. AB - A novel neural network has been devised that combines the advantages of cascade correlation and computational temperature constraints. The combination of advantages yields a nonlinear calibration method that is easier to use, stable, and faster than back-propagation networks. Cascade correlation networks adjust only a single unit at a time, so they train very rapidly when compared to back propagation networks. Cascade correlation networks determine their topology during training. In addition, the hidden units are not readjusted once they have been trained, so these networks are capable of incremental learning and caching. With the cascade architecture, temperature may be optimized for each hidden unit. Computational temperature is a parameter that controls the fuzziness of a hidden unit's output. The magnitude of the change in covariance with respect to temperature is maximized. This criterion avoids local minima, forces the hidden units to model larger variances in the data, and generates hidden units that furnish fuzzy logic. As a result, models built using temperature-constrained cascade correlation networks are better at interpolation or generalization of the design points. These properties are demonstrated for exemplary linear interpolations, a nonlinear interpolation, and chemical data sets for which the numbers of chlorine atoms in polychlorinated biphenyl molecules are predicted from mass spectra. PMID- 21644727 TI - Microelectrode control of surface-bound enzymatic activity. AB - Microelectrodes have been used to control the microscopically local reaction environment of surface-bound alcohol dehydrogenase (ADH). A surface comprised of agarose beads coated with immobilized ADH was prepared on a microscope slide and exhibited maximum activity toward the oxidation of ethanol, in the presence of nicotinamide adenine dinucleotide (NAD(+)), at a pH of 9.0. Microelectrode control of activity was achieved by submerging the slide in a solution of pH 6.0, well below the optimum value, and generating hydroxide at the microelectrode tip through the reduction of oxygen or water. An alkaline "sphere of influence" was set up around the microelectrode tip that, when positioned in close proximity to the enzyme surface, created a favorable reaction environment. The increased enzymatic activity was monitored by observing fluorescence of the reduced cofactor, NADH, using a fluorescence microscope equipped with an imaging camera. The fluorescent sphere diameter was characterized as a function of time, potential, and solution buffer strength. Optimum spatial resolution for enzymatic control was 7-12 MUm. PMID- 21644728 TI - Electrochemical and spectroscopic investigation of the reduction of dimethylglyoxime at mercury electrodes in the presence of cobalt and nickel. AB - Voltammograms (polarograms) obtained from solutions of cobalt and nickel containing dimethylglyoxime (dmgH(2)) are widely used for the trace determination of these metals. Detailed electrochemical and spectroscopic studies on the reduction process observed in the analytically important ammonia buffer media at mercury dropping, hanging, and pool electrodes are all consistent with an overall 10-electron reduction process, in which both the dmgH(2) ligand and cobalt ions are reduced in the adsorbed state: Co(II) + 2dmgH(2) ? (solution) [Co(II)(dmgH)(2)] + 2H(+); [Co(II)(dmgH)(2)] + Hg ? (electrode) [Co(II)(dmgH)(2)](ads)Hg; and [Co(II)(dmgH)(2)](ads)Hg + 10e(-) + 10H(+) -> Co(Hg) + 2[2,3-bis(hydroxylamino)butane]. The limited solubility of the nickel complex in aqueous media restricts the range of studies that can be undertaken with this system, but an analogous mechanism is believed to occur. Low temperature voltammetric studies in dichloromethane at a frozen hanging mercury drop electrode and in situ electron spin resonance electrochemical measurements on more soluble analogues of the dimethylglyoxime complexes are consistent with an initial one-electron reduction step being available in the absence of water. Deliberate addition of water to acetone solutions enables the influence of the aqueous environment on voltammograms and polarograms to be examined. The results of the present study are compared with the wide range of mechanisms proposed in other studies. PMID- 21644729 TI - Laser-induced fluorescence of se, as, and sb in an electrothermal atomizer. AB - Trace detection of Se, As, and Sb atoms has been performed by electrothermal atomization laser-induced fluorescence (ETA-LIF) approaches. Production of far-UV radiation necessary for excitation of As atoms at 193.696 nm and Se atoms at 196.026 nm was accomplished by stimulated Raman shifting (SRS) of the output of a frequency-doubled dye laser operating near 230 nm. Both wavelengths were obtained as second-order anti-Stokes shifts of the dye laser radiation and provided up to 10 MUJ/pulse, which was shown through power dependence studies to be sufficient for saturation in the ETA. An excited-state direct line fluorescence approach using excitation at 206.279 nm was also investigated for the LIF detection of Se. High-sensitivity LIF of Sb atoms was accomplished using 206.833-nm excitation and detection at 259.805 nm. The accuracy of the ETA-LIF approaches was demonstrated by determining the As and Se content of aqueous reference samples. The limits of detection (absolute mass) were 200 fg by ground-state LIF and 150 fg by excited state direct line fluorescence for Se, 200 fg for As, and 10 fg for Sb; these LODs compare favorably with results reported previously in the literature for ETA LIF, GFAAS, and ICP-MS methods. PMID- 21644730 TI - Measurement of SDS Micelle-Peptide Association Using (1)H NMR Chemical Shift Analysis and Pulsed-Field Gradient NMR Spectroscopy. AB - The binding of two simple tripeptides, glycyl-histidyl-glycine (GHG) and phenylalanyl-histidyl-phenylalanine (FHF) with SDS micelles was examined using (1)H NMR chemical shift analysis and self-diffusion coefficients measured with pulsed-field gradient NMR spectroscopy. The presence of GHG or FHF did not appear to significantly affect the critical micelle concentration (cmc) or the average size of the SDS micelles formed. The chemical shifts of several of the GHG resonances change as a function of SDS concentration, indicating an interaction between the peptide and the micelles. In addition, the concentration-dependent decrease observed for the GHG diffusion coefficients suggests association of the peptide with SDS micelles. The free and micelle-associated GHG are in fast exchange on both the (1)H chemical shift and diffusion time scales. The equilibrium constant for the binding of GHG to SDS micelles was determined from the analysis of the concentration dependence of the histidine C2 and C4 resonances to be 17 +/- 5 and 24 +/- 6 M(-)(1), respectively. The precision of the equilibrium constants obtained by analysis of the chemical shift data is limited by the small chemical shift changes observed. Analysis of the concentration dependence of the diffusion coefficients produced an equilibrium constant of 17 +/- 1 M(-)(1). The more hydrophobic peptide, FHF is strongly associated with the SDS micelles. Because the fraction of free FHF is small in these solutions, it was not possible to determine a formation constant for the interaction of FHF with the SDS micelles by analysis of either the (1)H chemical shift or diffusion coefficient data. The cmc of SDS in 0.10 M Na(2)C(2)O(4) buffer was determined to be 5.4 +/- 0.1 mM by analysis of the SDS diffusion coefficients in the absence of the peptides. The SDS cmc could also be extracted from the GHG and FHF diffusion coefficients measured as a function of the SDS concentration. The cmc determined from the GHG diffusion data, 5.7 +/- 0.2 mM, is in good agreement with the value determined from analysis of the SDS diffusion coefficients in the 5.0 mM GHG solution, 5.2 +/- 0.1 mM. The smaller cmc determined from the FHF diffusion data, 4.1 +/- 0.1 mM, may reflect some association of the SDS with the peptide prior to micelle formation in bulk solution. PMID- 21644731 TI - Order-sorting filters for a grating spectrometer and multichannel detection system: application to real-time spectroscopic ellipsometry. AB - A multichannel detection system with a grating spectrometer was employed for rapid spectroscopic ellipsometry measurements. Although a grating spectrometer is convenient for dispersing light, it suffers from overlapping orders. Here, we developed novel techniques to eliminate overlapping orders of spectra from a reflection grating. First, an optical filter is properly positioned in the measurement system. Second, a numerical filter is developed. A monochromatic source was used to deduce the exact amount of the overlapping orders, which were determined as the irradiance ratio of the second- to the first-order diffraction. The ratios were found to be increased from 5 to 11% in the 1.5-2.5 eV region. We used the values to correct for overlapping irradiance using the numerical filter. Finally, second-order reflection in real-time rotating element spectroellipsograms were corrected using the filters developed in this work. PMID- 21644732 TI - Characterization of a capillary zone electrophoresis/electrospray-mass spectrometry interface. AB - A dependable and stable CZE/ESI-MS interface has been constructed. To avoid instabilities in both, the capillary electrophoretic separation and the electrospray, the second of the three concentric capillaries in the three-layered sprayer has been replaced by an aluminum-coated fused-silica capillary with an inner diameter only slightly greater than the outer diameter of the separation capillary. By this means, the otherwise often observed destruction of the separation capillary ("electrodrilling") can be avoided completely due to the suppression of electrochemical processes leading to gas bubble formation at the tip of the sprayer. With some examples taken from different biochemical areas and by separation of natural compounds, the capability and the reliability of the modified sprayer as the central part of the interface are demonstrated. PMID- 21644733 TI - A micellar electrokinetic chromatographic method for determination of solubilization isotherms in surfactant micelles. AB - We have devised a new micellar electrokinetic chromatographic method for determination of the solubilization equilibrium constants of neutral solutes in surfactant micelles as functions of the intramicellar mole fraction of the solute. In this method, the running solution in a capillary and in electrode vessels contains the neutral solute as well as the surfactant micelles, and the injected solution contains the surfactant micelles and the dilute marker compounds for aqueous and micellar phases but not the solute. The mobility of the solute can be measured from a negative peak recorded on the chromatogram. The solute concentration in the capillary is established, and the mobility of the micellar phase incorporating the solute, which depends on the intramicellar mole fraction of the solute, can be measured from the migration time of the micelle marker. These merits enable precise determination of the solubilization isotherms. This method has been used to study the solubilization equilibria of benzene, phenol, and a series of aliphatic ketones in sodium dodecyl sulfate solution, and the validity and effectiveness of this method for quantifying solubilization have been verified. PMID- 21644734 TI - Electrokinetic chromatography using thermodynamically stable vesicles and mixed micelles formed from oppositely charged surfactants. AB - The electrokinetic chromatography (EKC) of a novel mixed surfactant system consisting of oppositely charged surfactants, sodium dodecyl sulfate (SDS) and n dodecyltrimethylammonium bromide (DTAB), was investigated. The chromatographic characteristics of large liposome-like spontaneous vesicles and rodlike mixed micelles formed from the mixture were explored and compared with those of SDS micelles. Separations of a series of n-alkylphenones showed that the spontaneous vesicles provided about a 2 times wider elution window than SDS micelles. Both vesicle and mixed micelle systems were found to provide larger methylene selectivity than SDS. The different elution order of a group of nitrotoluene geometric isomers with DTAB/SDS spontaneous vesicles and SDS micelles pseudostationary phases suggested the possibility of different separation mechanisms with these two systems. Comparisons of polar group selectivity, retention, and efficiency were made between vesicles, mixed micelles, and SDS micelles. The correlation between the logarithms of the retention factors (log k') and octanol-water partition coefficients (log P(ow)) for a group of 20 neutral compounds was also studied with DTAB/SDS vesicles. Spontaneous vesicles have great potential as a pseudostationary phase in electrokinetic chromatography. PMID- 21644735 TI - Study of interactions in supercritical fluids and supercritical fluid chromatography by solvatochromic linear solvation energy relationships. AB - Linear solvation energy relationships were used to study the retention process in supercritical fluid chromatography (SFC) and to gain a better understanding of intermolecular interactions in supercritical fluids. Correlation of SFC retention data with a set of solute solvatochromic parameters, which are also applicable to gas and liquid chromatography, yields information regarding the relative contributions of dispersion, cavity formation, dipolar, and hydrogen-bonding processes to retention. Dispersion interactions and cavity formation processes dominate retention on an open tubular poly(dimethylsiloxane) stationary phase with pure carbon dioxide as the mobile phase. Dipolar interactions and hydrogen bonding interactions are of decidedly less importance but do contribute significantly to retention. Based on prior solvatochromic studies of poly(dimethylsiloxane) and carbon dioxide, the changes in the regression coefficients with temperature and pressure are interpreted chemically. The relative importance of these contributions changes with temperature and pressure. As pressure increases, the carbon dioxide becomes more dense, and dispersion interactions between the solute and the mobile phase increase. A temperature increase at constant pressure decreases dispersion interactions with the stationary phase, as in gas chromatography, but also decreases dispersion interactions with the mobile phase, due to a decrease in carbon dioxide density. On the basis of the solvatochromic coefficients, carbon dioxide acts as both a Lewis base and a Lewis acid. The quality of fit for these correlations is very high and compares favorably with similar studies in gas chromatography and liquid chromatography, permitting the prediction of retention behavior from a solute's solvatochromic parameters. PMID- 21644736 TI - Capillary affinity gel electrophoresis for combined size- and sequence-dependent separation of oligonucleotides. AB - An interesting new approach to capillary affinity gel electrophoresis (CAGE) has been developed for the selective capture and separation of homopolymer and heteropolymer oligonucleotides. The combination of selectivity of bioaffinity recognition and high-resolution power of capillary gel electrophoresis allows the on-line sequence- and size-specific separation of oligonucleotides. Both rigid gel formulations and viscous replaceable polymer solutions having user-defined, single-stranded oligonucleotides covalently attached as recognition sequences are used. Contrary to most known affinity systems in capillary electrophoresis, which operate in a continuous mode, binding and release are accomplished in two steps, effectively separating the affinity from the separation step. At low temperature, oligonucleotides with complementary sequences in the analyte solution will bind to the immobilized recognition sequence while unrelated oligonucleotides will continue to migrate. This step is a preseparation, removing all nonspecific solutes from the sample. The release of the bound solutes is achieved at elevated temperature, allowing a probe of cross-reactivity for a given biorecognition element. Applications for high-resolution separations of short oligonucleotides and their mismatches are shown, and the potential for on-line preconcentration and separation of dilute analyte solutions, thus effectively enhancing the sensitivity, is demonstrated. PMID- 21644737 TI - Selective extraction of pesticides from lipid-containing matrixes using supercritical binary gas mixtures. AB - When supercritical carbon dioxide is used for extraction of trace compounds from lipid-rich samples, the resulting extract can contain a substantial amount of lipid coextractive which interferes in the subsequent chromatographic analysis. In this case, a cleanup step, which is time-consuming, is required in order to remove or reduce the lipid content in the extract. In this study, we report on a new approach for the extraction of trace compounds which significantly reduces the quantity of lipid coextractives, thereby eliminating the need for a sample cleanup step. An autoclave high-pressure reactor equipped with a mechanical stirring device was used for mixing gases, such as carbon dioxide and nitrogen, to generate the desired fluid mixtures. The composition of the mixtures could be adjusted by two mass flow controllers prior to the autoclave mixing device. The generated carbon dioxide/nitrogen binary mixtures were then employed to facilitate supercritical fluid extraction (SFE) at high pressures and elevated temperatures. The pressurized CO(2)/nitrogen binary fluid mixture demonstrated sufficient solvation power for quantitative recoveries of trace fortified organophosphorus and incurred organochlorine pesticides from poultry fat while significantly reducing lipid solubility in the fluid. This allowed the development of a method that produced an extract with minimal lipid content that could be used directly for gas chromatographic analysis, thereby eliminating the need to clean-up the extract. PMID- 21644738 TI - Selective HPLC Analysis of n-Alkyl Hydroperoxides up to C(18)H(38)O(2). AB - A series of n-alkyl hydroperoxides are separated by HPLC and detected by their postcolumn reaction with horseradish peroxidase and p-hydroxyphenylacetic acid (HPAA) to yield a fluorescent product; several secondary and tertiary hydroperoxides, some 1-hydroxyalkyl hydroperoxides, and a few branched hydroperoxides are also examined. n-Alkyl hydroperoxides up to at least C-18 react with the enzyme with only minimally reduced efficiency at greater alkyl chain length. The effects of the column, the eluent, and the pH of the sample reaching the detector are described. The detection limit with gradient elution ranges from 0.4 MUmol L(-)(1) for n-hexyl hydroperoxide to 1 MUmol L(-)(1) for n octadecyl hydroperoxide. PMID- 21644739 TI - Differential detection of enantiomeric gaseous analytes using carbon black-chiral polymer composite, chemically sensitive resistors. AB - Carbon black-chiral polymer composites were used to provide diagnostic differential resistance responses in the presence of enantiomers of chiral gaseous analytes. Vapors of (+)-2-butanol and (-)-2-butanol, (+)-alpha-pinene and (-)-alpha-pinene, (+)-epichlorohydrin and (-)-epichlorohydrin, and methyl (+)-2 chloropropionate and methyl (-)-2-chloropropionate were generated and passed over a chemically sensitive carbon black-poly((R)-3-hydroxybutyrate-co-(R)-3 hydroxyvalerate) (77% butyrate) composite resistor. Each enantiomer of a pair produced a distinct relative differential resistance change on the chiral detector, whereas both enantiomers of a set produced identical signals on achiral carbon black-poly(ethylene-co-vinyl acetate) (82% ethylene) detectors. PMID- 21644740 TI - Single-molecule identification in flowing sample streams by fluorescence burst size and intraburst fluorescence decay rate. AB - We report a multiplex technique for identification of single fluorescent molecules in a flowing sample stream by correlated measurement of single-molecule fluorescence burst size and intraburst fluorescence decay rate. These quantities were measured simultaneously for single fluorescent molecules in a flowing sample stream containing a dilute mixture of fluorescent species: Rhodamine 6G and tetramethylrhodamine isothiocyanate. Using a detailed Monte Carlo simulation of our experiment, we calculate single-molecule detection efficiencies and confidence levels for identification of these species and identify major sources of error for single-molecule identification. The technique reported here is applicable to distinguishing between fluorophores with similar spectroscopic properties and requires only a single excitation wavelength and single fluorescence emission detection channel. PMID- 21644741 TI - Counting single chromophore molecules for ultrasensitive analysis and separations on microchip devices. AB - Separations of 15 pM rhodamine 6G and 30 pM rhodamine B performed in a micromachined electrophoresis channel were detected by counting fluorescence bursts from individual molecules. The migration times, peak widths, and analyte concentrations were estimated from the number and the migration time distribution of the detected molecules. Concentration detection limits estimated at >99% confidence were 1.7 pM rhodamine 6G and 8.5 pM rhodamine B. The separations required <35 s and the relative migration time uncertainties were less than 2.0%. These are the lowest detection limits reported for microchip separation devices and the first example of single-chromophore molecular counting for detection of chemical separations. PMID- 21644742 TI - A quadrupole ion trap with cylindrical geometry operated in the mass-selective instability mode. AB - A cylindrical geometry ion trap is used to record mass spectra in the mass selective instability mode. The geometry of the cylindrical ion trap has been optimized to maximize the quadrupole field component relative to the higher-order field content through field calculations using the Poisson/Superfish code and through experimental variation of the electrode structure. The results correspond well with predictions of the calculations. The trap has been used to record mass spectra with better than unit mass resolution, high sensitivity, and a mass/charge range of ~600 Th. Multistage (MS(3)) experiments have been performed, and the Mathieu stability region has been experimentally mapped. The performance of this device compares satisfactorily with that of the hyperbolic ion trap. PMID- 21644743 TI - A phase boundary potential model for apparently "twice-nernstian" responses of liquid membrane ion-selective electrodes. AB - A model that describes divalent cation responses of liquid membrane ion-selective electrodes based on acidic ionophores and ionic sites is presented. Response slopes for membranes with ionophore and anionic sites are predicted to change from Nernstian to apparently "twice-Nernstian" and then back to Nernstian again as the pH of the sample solution decreases. A maximum measuring range for apparently "twice-Nernstian" responses is expected for membranes with 50 mol % anionic sites relative to the ionophore. On the other hand, membranes with ionophore and cationic sites are expected to give only Nernstian responses, either to divalent cations at high pH or to H(+) at low pH. The validity of the present model has been confirmed experimentally with the two Ba(2+)-selective carboxylate ionophores monensin and lasalocid and the Ca(2+)-selective organophosphate ionophore bis(2-heptylundecyl) phosphate. Addition of anionic sites gave apparently "twice-Nernstian" slopes for monensin at pH 7.0 (56.6 mV/decade), for lasalocid at pH 4.0 (53.3 mV/decade), and for bis(2 heptylundecyl) phosphate at pH 3.5 (53.6 mV/decade). Membranes with cationic sites showed only pH responses at the respective pH. The apparently "twice Nernstian" responses as discussed here are the first examples of super-Nernstian responses that can be explained with a quantitative model based on thermodynamic equilibria. PMID- 21644744 TI - Fabrication of Boron-Doped CVD Diamond Microelectrodes. AB - Diamond microelectrodes are fabricated using microwave plasma CVD for the growth of electrically conducting single microcrystallite diamonds as well as diamond films on etched tungsten wires which are subsequently sealed in glass. The electroactive diamond is exposed by either mechanical polishing or by chemical etching of the glass. The resulting microelectrodes yield steady-state cyclic voltammograms at low scan rates. PMID- 21644745 TI - A Ring-Disk Study of the Competition between Anodic Oxygen-Transfer and Dioxygen Evolution Reactions. AB - Voltammetric data obtained at a rotated ring-disk electrode for oxidation of dimethyl sulfoxide (DMSO) to dimethyl sulfone support the conclusion that anodic O-transfer and O(2)(aq)-evolution reactions are in competition for a common reaction precursor. That species is concluded to be the adsorbed OH radical generated by anodic discharge of H(2)O. Because of this competition, an increase in the flux of the reactant in the O-transfer reaction causes a decrease in the rate of O(2)(aq) evolution. Therefore, difference voltammetry and hydrodynamically modulated voltammetry cannot be considered reliable for accurate deconvolution of the total electrode current resulting from these concomitant processes. A simple mathematical model is developed to describe the total current resulting from these simultaneous processes, and results of preliminary tests of this model are in good agreement with experimental data for DMSO oxidation. PMID- 21644746 TI - Development and calibration of field-effect transistor-based sensor array for measurement of hydrogen and ammonia gas mixtures in humid air. AB - A sensor array for analyzing hydrogen and ammonia gas mixtures in humid air has been developed, built into a rugged system, and calibrated for laboratory testing. The sensor array is comprised of four chemically sensitive field-effect transistors (CHEMFETs). Chemically sensitive layers for the sensors were developed and tested using a Kelvin probe. A combination of catalytic and noncatalytic thin layers (palladium and polyaniline) was selected for the four sensor array. The work function responses of the CHEMFET sensor array to mixtures of hydrogen, ammonia, and humid air were measured. Chemometric multivariate methods, linear and nonlinear partial least squares, were used for the calibration of the sensor array using gas mixtures in the concentration range from 0 to 10 000 ppm hydrogen and ammonia in humid air. The sensor array for ammonia showed good sensitivity, selectivity, response time, and stability and is recommended for field deployment. In contrast, the sensor array for hydrogen, though highly sensitive to hydrogen, demonstrated inadequate stability, requiring further development before deployment is recommended. PMID- 21644747 TI - A graphite furnace electrothermal vaporization system for inductively coupled plasma atomic emission spectrometry. AB - An improved graphite furnace electrothermal vaporization device equipped with an autosampler for inductively coupled plasma atomic emission spectrometry is presented. The transport losses of eight selected analytes in the individual segments of the device were determined by means of the radiotracer technique by applying amounts traced comparable to those to be determined in real samples. The results obtained from the radiotracer study were the basis for further improvement of the interface design, leading to considerable increase of the total transport efficiency, which finally was found to be between 26 (for Cr) and 57% (for Ga). The whole system consists of a graphite furnace vaporizer, a power supply, a gas flow box, and an autosampler with incorporated microbalance. The temperature program, gas flows, and autosampler functions are controlled by a data station which also provides the data acquisition and processing of the transient signals. The performance parameters of the developed system were evaluated using aqueous standard solutions. Absolute limits of detection for most analytes were between 0.1 and 1 ng, and for As, K, Ni and Pb, they were between 2 and 3.2 ng. PMID- 21644748 TI - Analysis of Carbendazim in Agricultural Samples by Laser Desorption and REMPI Time-of-Flight Mass Spectrometry. AB - A method has been developed for the analysis of carbendazim in agricultural samples. This has been accomplished by the combination of laser desorption with resonance-enhanced multi-photon ionization (REMPI) coupled to time-of-flight mass spectrometry detection. After the optimization of the experimental conditions and the location of the resonant wavelength of the substance, several samples of pepper extract enriched with carbendazim were analyzed, finding a detection limit of the same order of magnitude as that of current GC and HPLC techniques, but with higher sensitivity and faster sample preparation. PMID- 21644749 TI - Microprobe speciation analysis of inorganic solids by fourier transform laser mass spectrometry. AB - Fourier transform (FT) laser microprobe mass spectrometry (LMMS) aims at the characterization of local constituents at the surface of solids. Signals from structural fragments specify the main building blocks of the analyte while adduct ions, consisting of one or two intact analyte molecules and a stable ion, allow specific identification of the molecule. A series of inorganic reference compounds including binary salts, oxides, and oxy salts was analyzed to assess the FT LMMS capabilities for the determination of the inorganic molecular composition. Compounds from different classes can be tentatively identified by deductive reasoning while those with the same elements in different stoichiometries require comparison with reference spectra. PMID- 21644750 TI - Microplasma mass spectrometric detection in capillary gas chromatography. AB - A simple and miniaturized 350-kHz helium discharge for plasma mass spectrometric detection in gas chromatography (GC) has been developed. The plasma was sustained at low pressure within the end of the capillary GC column (0.32-mm i.d.) inside the ion source housing of a quadrupole mass spectrometer. This allowed direct introduction of ions from the plasma to the mass analyzer using only a repeller and electrostatic lenses to focus the ions. The plasma was sustained in only 25 mL min(-)(1) of helium, which was accepted by the mass spectrometer vacuum system. This low gas flow also served to enhance the energy density of the discharge and to produce a narrow spray of ions toward the mass analyzer. Due to the miniaturized nature of the plasma, it was operated at a low power level (2.0 W), and traces of oxygen were added to avoid deposition of carbon. With this new concept for GC plasma mass spectrometric detection, chlorine was successfully monitored down to the 2.2 pg s(-)(1) level without interference from elements like C, S, P, O, F, and N. PMID- 21644751 TI - Large Molecule Characterization Based upon Individual Ion Detection with Electrospray Ionization-FTICR Mass Spectrometry. AB - We report a new method for mass spectrometric measurements of high-molecular weight species based on the summation of sequential Fourier transform ion cyclotron resonance (FTICR) spectra of individual multiply charged ions. This approach produces statistically useful mass spectra for large multiply charged molecular species formed by electrospray ionization and circumvents conventional limitations upon achievable resolving power and precision for high-molecular weight species which arise due to Coulombic constraints. For very large molecules with tens to thousands of charges each, the total number of charges required to define the charge-state distribution, and thus provide accurate mass information, greatly exceeds the useful charge capacity of the FTICR cell. As trapped ion populations approach or exceed this capacity, FTICR performance degrades due to large frequency shifts, peak coalescence phenomena, and rapid loss of ion packet coherence, which effectively precludes high-resolution and precision measurements for molecules above ~80-kDa size for a 7-T magnetic field strength. The present approach is based on the summation of many spectra having moderate populations of individual ions and relies on sensitivity sufficient for individual ion detection. While the number of trapped ions contributing to each mass spectrum may generally be insufficient to define the isotopic or charge-state distributions (and thus produce accurate information on the molecular weight distribution in a conventional fashion), the present data processing and summation approach suppresses the noise component (as well as smaller signals) that would otherwise be problematic. Importantly, this approach circumvents natural limitations for very high molecular weight species due to Coulombic interactions and thus provides a basis for much greater resolution and mass measurement accuracy than otherwise possible. This paper presents the details of this approach and its demonstration for the 66-kDa protein bovine serum albumin (where the conventional approach is also feasible) and discusses important aspects of the data manipulation. PMID- 21644752 TI - Cs(+) Speciation on Soil Particles by TOF-SIMS Imaging. AB - Soil particles exposed to CsI solutions were analyzed by imaging time-of-flight secondary ion mass spectrometry and also by scanning electron microscopy/energy dispersive X-ray spectroscopy (SEM/EDS). The results showed that Cs(+) could be detected and imaged on the surface of the soil particles readily at concentrations down to 160 ppm, which corresponds to 0.04 monolayer. Imaging revealed that most of the soil surface consisted of aluminosilicate material. However, some of the surface was more quartzic in composition, primarily SiO(2) with little Al. It was observed that adsorbed Cs(+) was associated with the presence of Al on the surface of the soil particles. In contrast, in high SiO(2) areas of the soil particle where little Al was observed, little adsorbed Cs(+) was observed on the surface of the soil particle. Using EDS, Cs(+) was observed only in the most concentrated Cs(+)-soil system, and Cs(+) was clearly correlated with the presence of Al and I. These results are interpreted in terms of multiple layers of CsI forming over areas of the soil surface that contain substantial Al. These observations are consistent with the hypothesis that the insertion of Al into the SiO(2) lattice results in the formation of anionic sites, which are then capable of binding cations. PMID- 21644753 TI - Modeling of the impact of ionic strength on the electroosmotic flow in capillary electrophoresis with uniform and discontinuous buffer systems. AB - A new dynamic computer model permitting the combined simulation of the temporal behavior of electroosmosis and electrophoresis under constant voltage or current conditions and in a capillary which exhibits a pH-dependent surface charge has been constructed and applied to the description of capillary zone electrophoresis, isotachophoresis, and isoelectric focusing with electroosmotic zone displacement. Electroosmosis is calculated via use of a normalized wall titration curve (mobility vs pH). Two approaches employed for normalization of the experimentally determined wall titration data are discussed, one that considers the electroosmotic mobility to be inversely proportional to the square root of the ionic strength (method based on the Gouy-Chapman theory with the counterion layer thickness being equal to the Debye-Huckel length) and one that assumes the double-layer thickness to be the sum of a compact layer of fixed charges and the Debye-Huckel thickness and the existence of a wall adsorption equilibrium of the buffer cation other than the proton (method described by Salomon, K.; et al. J. Chromatogr. 1991, 559, 69). The first approach is shown to overestimate the magnitude of electroosmosis, whereas, with the more complex dependence between the electroosmotic mobility and ionic strength, qualitative agreement between experimental and simulation data is obtained. Using one set of electroosmosis input data, the new model is shown to provide detailed insight into the dynamics of electroosmosis in typical discontinuous buffer systems employed in capillary zone electrophoresis (in which the sample matrix provides the discontinuity), in capillary isotachophoresis, and in capillary isoelectric focusing. PMID- 21644754 TI - Contactless conductivity detection for capillary electrophoresis. AB - A contactless capacitively coupled conductivity detector for capillary electrophoresis is introduced. The detector consists of two electrodes which are placed cylindrically around the outer polyimide coating of the fused-silica capillary with a detection gap of 2 mm. The electrodes form a cylindrical capacitor, and the electric conductivity of the solution in the gap between the electrodes is measured. A high audio or low ultrasonic frequency for coupling of the ac voltage is used in order to minimize the influence of reactance of the liquid. For an improved version of the detector, two syringe cannulas are used as the electrodes and the capillary is simply assembled into the tubing. This allows an easy placement of the detector on various positions along the capillary. The limit of detection of inorganic cations and anions is 200 ppb, as determined for sodium and chloride, respectively. PMID- 21644755 TI - High-Resolution Studies of Hyaluronic Acid Mixtures through Capillary Gel Electrophoresis. AB - Hyaluronic acid is a negatively charged polysaccharide with a high degree of polydispersity that makes the separation of its oligomers extremely difficult. Through the use of columns filled with a highly viscous polyacrylamide matrix, the unit resolution of hyaluronate oligomers could be achieved, up to at least 80 kDa of mass, through capillary electrophoresis. As analytical application examples, the fractions of enzymatically or ultrasonically degraded hyaluronates were monitored through this method. Because of the very high resolving power, peaks additional to the regular oligomers can be observed that are assumed to be conformers of this regular, unbranched biopolymer. PMID- 21644756 TI - A Family of Single-Isomer Chiral Resolving Agents for Capillary Electrophoresis. 3. Heptakis(2,3-dimethyl-6-sulfato)-beta-cyclodextrin. AB - The third member of a new family of single-isomer charged cyclodextrins, the sodium salt of heptakis(2,3-dimethyl-6-sulfato)-beta-cycldextrin, has been synthesized, characterized, and used for the capillary electrophoretic separation of the enantiomers of neutral, acidic, basic, and zwitterionic analytes. Though heptakis(2,3-dimethyl-6-sulfato)-beta-cyclodextrin complexes much less strongly with any of the analytes tested here than the previously synthesized heptakis(2,3 diacetyl-6-sulfato)-beta-cyclodextrin and heptakis-6-sulfato-beta-cyclodextrin, it offers excellent enantioselectivities, complementary to those of the other two single-isomer, differently functionalized charged cyclodextrins. Confirming the predictions of the charged resolving agent migration model, heptakis(2,3-dimethyl 6-sulfato)-beta-cyclodextrin allowed for the reversal of the migration order of the enantiomers of neutral analytes as the cyclodextrin concentration was increased. Just as with the previous two single-isomer charged resolving agents, separation selectivity for the acidic, basic, and zwitterionic analytes could increase, decrease, or pass a maximum as the cyclodextrin concentration was increased, depending on the respective binding strength of the enantiomers and the ionic mobilities of both the complexed and noncomplexed forms of the enantiomers. PMID- 21644757 TI - Quantitative Measurements in Continuous-Flow HPLC/NMR. AB - Two methods for the quantitative determination of compounds in continuous-flow HPLC/NMR are described. The first method uses an internal standard (caffeine) of known concentration directly mixed into the mobile phase, while with the second method, a known amount of internal standard is injected onto the column during the chromatographic run. The latter method was validated using several nitroaromatic compounds and explosives. Deviations between the injected and calculated amounts of analytes are usually below 10% while the relative standard deviation ranges from 2% in the upper microgram range to 40% at the limit of detection. PMID- 21644758 TI - Deuterium nuclear magnetic resonance spectroscopy as a probe for reversed-phase liquid chromatographic bonded-phase solvation. 3. Tetrahydrofuran and water binary systems. AB - Deuterium nuclear magnetic resonance ((2)H NMR) spectroscopy is a useful tool in probing molecular association between components in a binary hydro-organic mixture, such as tetrahydrofuran (THF) and water. Measurements of (2)H longitudinal relaxation times (T(1)) for deuterated analogs of THF and water provide this type of information, since the magnitude of T(1) is indicative of molecular association and mobility. This technique is also useful in determining the interactions between the mobile phase and reversed-phase stationary phase. T(1) measurements of slurry samples, consisting of both mobile- and stationary phase components, allow for changes in the longitudinal relaxation time of mobile phase components upon contact with the stationary phase (DeltaT(1)) to be monitored. These DeltaT(1) values are indicative of the degree of association of the THF-water mobile-phase components with the stationary phase. PMID- 21644759 TI - Participation of Cluster Species in the Solvation Mechanism of a Weak Polar Solute in a Methanol/Water Mixture over a 0.2-0.7 Water Fraction Range: High Performance Liquid Chromatography Study. AB - It is known that a methanol/water mixture has a quaternary organization consisting of free water W, free methanol M, and two types of methanol/water clusters. A 1:1 methanol/water cluster has been observed at a high methanol concentration, and a 5:1 cluster has been observed at a low methanol concentration. In the water fraction range used in this paper, 0.2-0.7, the number of MW(5) clusters was always inferior to the number of MW clusters. When a weak polar solute is introduced into such a mixture, it is preferentially solvated by free methanol and the MW water clusters. Using this mixture as the mobile phase, a novel mathematical theory is presented to describe, in this water fraction range, the variations of the retention factor k' of alkyl benzoate esters and benzodiazepines in reversed-phase liquid chromatography. Excellent predictions of k' versus free methanol and methanol/water fractions were obtained. For the first time, using this model, enthalpy, entropy, and the Gibbs free energy of the two solute solvation processes were evaluated. Enthalpy entropy compensation revealed that the main parameters determining retention in the range of the water fraction (0.2-0.7) increased as follows: free methanol < > solute solvation > methanol/water cluster <-> solute solvation > RP18 stationary phase <-> solute interaction. These results agree well with values obtained when ACN was used instead of methanol. PMID- 21644760 TI - Extraction of petroleum hydrocarbons from soil using supercritical argon. AB - The use of supercritical argon is described for the extraction of petroleum hydrocarbons from soil samples. Argon is an attractive solvent because it is inexpensive and inert. Additionally, it has a clear spectral window in the infrared region which makes it useful for on-line (i.e., directly coupled) experiments. Spiking studies conducted with gasoline, no. 1 fuel oil, and no. 5 fuel oil on sand, loam, and clay show that component recovery rates for argon supercritical fluid extraction (SFE) generally increase with increasing pressure and/or temperature. The highest recovery rates (and recoveries) were obtained for Ar SFE at 500 atm and 150 degrees C. Under these conditions, the components of the gasoline and no. 1 fuel oil spikes could be recovered in as little as 12 min. However, the no. 5 fuel oil components could not be quantitatively removed from the loam and clay matrixes even for extraction times as long as 100 min. We also show in this work that Ar SFE performs similarly to CO(2) SFE for petroleum hydrocarbon contamination in real-world soil samples under moderate pressure and temperature conditions. Specifically, Ar SFE and CO(2) SFE have similar recoveries and reproducibilities, but Ar SFE requires a slightly longer extraction time. PMID- 21644761 TI - Assay system for the herbicide 2,4-dichlorophenoxyacetic Acid using a molecularly imprinted polymer as an artificial recognition element. AB - Noncovalent molecular imprinting of a synthetic polymer with the herbicide 2,4 dichlorophenoxyacetic acid has been achieved in the presence of the polar solvents methanol and water. Formation of the prearranged complex relied on hydrophobic and ionic interactions between the template and the functional monomer 4-vinylpyridine. The polymer obtained binds the original template with an appreciable selectivity over structurally related compounds. The potential use of micrometer-sized imprinted polymer particles as the recognition element in a radioligand binding assay for 2,4-dichlorophenoxyacetic acid is demonstrated. PMID- 21644762 TI - Open-ended photoacoustic cells: application to two-layer samples using pulse laser-induced photoacoustics. AB - To measure the difference between samples in situ, two types of open-ended photoacoustic cells were constructed. One type was a differential open-ended photoacoustic cell with a differential microphone, which connected two equivalent open-ended cell chambers through two waveguides. The other design used single open-ended photoacoustic cell chambers, each with an electric condenser microphone. A pair of these was used to suppress background signal by subtraction of the signals. The performance of these cells was compared to that of a conventional photoacoustic apparatus with a chopper. Detection limits of Sudan red for those cells were about 1.29 and 2.35 ng for a differential open-ended cell and a single open-ended cell, respectively. The photoacoustic cells were then applied to the pulse mode operation, using a nitrogen laser as a pulse light source. The laser beams were led to the photoacoustic cells through quartz optical fibers. Using this instrument setup, depth profiling analysis was carried out for two-layer samples made of poly(methyl methacrylate) (PMMA) or polystyrene (PS) films. A linear relationship was obtained between the signal delay time and the film thickness. The delay time of the signal (tau) was expressed in terms of the thermal diffusivity (kappa/cm(2) s(-)(1)) of the film as tau (ms) ? (2.25 * 10(-3)) x (MUm) kappa(-1/2). The method was successful in measuring the thickness of transparent surface layers. We applied this technique to observe the generation of a certain chromophore, thought to be melanin, under the surface of human skin tanned by exposure to long-wavelength UV light. PMID- 21644763 TI - Confinement and manipulation of individual molecules in attoliter volumes. AB - We report observation of fluorescence from individual rhodamine 6G molecules in streams of charged 1-MUm-diameter water droplets. With this approach, illumination volumes comparable to diffraction-limited fluorescence microscopy techniques (<=500 aL) are achieved, resulting in similarly high contrast between single-molecule fluorescence signals and nonfluorescent background. However, since the fluorescent molecules are confined to electrically charged droplets, in situ electrodynamic manipulation (e.g., focusing, switching, or merging) can be accomplished in a straightforward manner, allowing experimental control over both the delivery of molecules of interest to the observation region and the laser molecule interaction time. As illustrated by photocount statistics that are independent of molecular diffusion and spatial characteristics of the excitation field, individual rhodamine 6G molecules in 1-MUm droplets are reproducibly delivered to a target a few micrometers in diameter at a rate of between 10 and 100 Hz, with laser beam transit times more than 1 order of magnitude longer than diffusion-limited laser-molecule interaction times in equivalent volumes of free solution. PMID- 21644764 TI - Templated pores in hydrogels for improved size selectivity in gel permeation chromatography. AB - The pore structures of cross-linked polyacrylamide gels can be altered by polymerizing in the presence of high concentrations of unreactive, micellar surfactant cosolutes which act as "templates". Removal of surfactant after polymerization is expected to leave pores with the approximate shape and dimensions of the surfactant micelles. A simple model was developed to simulate gel permeation chromatography (GPC) separations of globular proteins on templated gels. The model assumes that the partition coefficient for sieving of a protein is equal to the fraction of gel volume accessible to a sphere with a radius equal to the protein Stokes radius. The total gel volume is considered to include a fraction that is a conventional, random gel matrix and a remaining fraction contributed by templated pores. The pore size distribution of the conventional gel was estimated using the Ogston equation, which approximates the matrix as a random collection of long, thin, rigid fibers. Templated pores were assumed to have a Gaussian distribution of radii centered about some mean determined by the micelle radius. In comparison to conventional media, gels with templated pores are predicted to exhibit more sharply defined exclusion limits and improved resolution over a narrow size range centered on the mean templated pore size. Selectivity and resolution are expected to increase as the volume fraction of templated pores is increased and as the dispersion of templated pore radius is decreased. Small changes in template radius lead to large changes in the molecular weight range of optimal separation of globular proteins. It should be possible to create a series of GPC media that collectively offer high resolution over the molecular weight range of most globular proteins of interest. PMID- 21644765 TI - Hydrophilic interaction chromatography-electrospray mass spectrometry analysis of polar compounds for natural product drug discovery. AB - For the drug discovery efforts currently taking place within the pharmaceutical industry, natural product extracts have been found to provide a valuable source of molecular diversity which is complementary to that provided by traditional synthetic organic methods or combinatorial chemistry. However, there exists a need for analytical tools that can facilitate the separation and characterization of components from these sources in a rapid manner. Specifically, the evaluation of highly polar compounds (i.e., compounds that cannot be retained on traditional reversed-phase stationary phases) has been challenging, and a hydrophilic interaction chromatography-electrospray ionization mass spectrometry (HILIC-ESI MS) method was developed to meet this need. In this investigation, amide-, Polyhydroxyethyl Aspartamide-, and cyclodextrin-based packings provided superior performance for the analysis of a set of polar natural product compounds. The properties of the mobile-phase buffers also greatly impacted the separations, and relative to other volatile buffering agents, ammonium acetate at a concentration of approximately 6.5 mM was determined to facilitate optimal HILIC retention, reproducibility, and durability. An optimized HILIC-ESI-MS system was successfully applied for the analysis of complex natural product mixtures. The techniques described in this report should also prove useful for the analysis of polar compounds from synthetic sources of molecular diversity such as combinatorial chemistry. PMID- 21644766 TI - Imaging of Pressure- and Electrokinetically Driven Flows through Open Capillaries. AB - A new tool for imaging both scalar transport and velocity fields in liquid flows through microscale structures is described. The technique employs an ultraviolet laser pulse to write a pattern into the flow by uncaging a fluorescent dye. This is followed, at selected time delays, by flood illumination with a pulse of visible light which excites the uncaged dye. The resulting fluorescence image is collected onto a sensitive CCD camera. The instrument is designed as an oil immersion microscope to minimize beam steering effects. The caged fluorescent dye is seeded in trace quantities throughout the active fluid, thus images with high contrast and minimal distortion due to any molecular diffusion history can be obtained at any point within the microchannel by selectively activating the dye in the immediate region of interest. We report images of pressure- and electrokinetically driven steady flow within round cross section capillaries having micrometer scale inner diameters. We also demonstrate the ability to recover the velocity profile from a time sequence of these scalar images by direct inversion of the conserved scalar advection-convection equation. PMID- 21644767 TI - Transient isotachophoretic-electrophoretic separations of lanthanides with indirect laser-induced fluorescence detection. AB - Indirect laser-induced fluorescence was used for the detection of several lanthanide species separated by capillary electrophoresis. Quinine sulfate was the fluorescent component of the background electrolyte, and alpha hydroxyisobutyric acid was added as a complexing agent to enable the separation of analyte ions that have similar mobilities. The UV lines (333-364 nm) of an argon ion laser were used as the excitation source with a diode array detector for monitoring the fluorescent emission at 442 nm. Electrokinetic injections and transient isotachophoresis were implemented to stack the analyte ions into more concentrated zones. On-line preconcentration factors were determined to be ~700 and resulted in limits of detection for La(3+), Ce(3+), Pr(3+), Nd(3+), Sm(3+), and Eu(3+) in the low-ppb range (6-11 nM). PMID- 21644768 TI - Stepper motor controlled micro gas valve inlet system for gas chromatography. AB - A novel gas valve inlet system for gas chromatography is described. The device incorporates a capillary sample gas delivery tube containing a small orifice in its side from which sample vapor continuously flows. A precision micro stepper motor is used to translate the sample delivery tube parallel to its axis so that the orifice passes by the end of the fused silica separation column. The inlet end of the column and the sample delivery tube are housed in a pressurized injection port which also contains purge flows to vent sample between injections. Two operating modes are described. In the sweep mode, the orifice sweeps past the column end at a constant, adjustable velocity. In the park mode, the orifice is parked in front of the column end for a software selectable time. Injection sample size and bandwidth are adjustable. Bandwidths (sigma) as small as a few milliseconds make the inlet suitable for high-speed gas chromatography as well as conventional GC. PMID- 21644769 TI - Potentiometric detection of alkali and alkaline Earth metal cations in capillary electrophoresis with simplified electrode alignment and enhanced separation and sensitivity. AB - Coated-wire electrodes were employed as potentiometric detectors for the analysis of alkali and alkaline earth metals by capillary electrophoresis. Easy electrode alignment was achieved by a Plexiglas capillary-electrode holder, which makes it unnecessary to employ a microscope and a micromanipulator. The addition of a crown ether to the running electrolyte not only renders possible the complete separation of ammonium and potassium but also improves considerably the sensitivity of the coated-wire electrode. We studied the dependence of the migration times on the crown ether concentration as well as the change in sensitivity by means of calibration curves for potassium and calcium. PMID- 21644770 TI - Multiple labeling of proteins. AB - Fluorescent dyes are often used to label proteins before analysis by capillary electrophoresis. Fluorescent labeling produces spectacular improvements in sensitivity compared with UV absorbance detection of the native protein. However, labeling of the protein can lead to significant band broadening. This band broadening is interpreted as a result of multiple labeling of the protein, wherein one or more fluorescent molecules are bound to the protein. The heterogeneous reaction products, which are presumed to have different mobilities, generate a broad peak during electrophoresis. There has been little direct evidence for multiple labeling as the cause of band broadening of proteins. In this paper, we perform electrophoresis on native green fluorescence protein, along with the reaction products produced by fluorescence labeling. For short incubations, a series of regularly spaced components are resolved by free-zone electrophoresis; upon longer incubation, the product peaks merge together, forming a broad envelope. PMID- 21644771 TI - Cyclic voltammetric detection in capillary electrophoresis with application to metal ions. AB - Fast cyclic voltammetry (CV) was evaluated over sweep rates of 20-1000 V/s at Au disk electrodes (25 and 10 MUm) for end-capillary detection in capillary electrophoresis with metal ions as test analytes; some studies were also done with 25-MUm Pt disk electrodes. The waveform applied to the electrode consisted of a preconcentration period (55-330 ms) followed by cyclic voltammetry (2-100 ms). Maximum signal-to-noise was obtained with the integrated CV current as the analytical signal, and this was linearly proportional to sweep rate; maximum response was obtained at sweep rates of >100 V/s for 10-MUm electrodes and >200 V/s for 25-MUm electrodes; sweep rates of >400 V/s caused peak tailing due to trapping of the analyte at the electrode. With this CV detection approach, comigrating analytes could be identified and determined. Reproducibilities for six analytes over the range 1.0 * 10(-)(7)-1.0 * 10(-)(5) mol/L were 2%-5%, and calibration curves were linear, with response factors in the range of 2%-6%. Detection limits (2 * peak-to-peak baseline noise) were in the range of 5 * 10( )(9)-4 * 10(-)(8) mol/L, which are 1-2 orders of magnitude better than results obtained previously with square-wave pulsed amperometric detection of metal ions. PMID- 21644772 TI - Solid-phase microextraction and headspace solid-phase microextraction for the determination of polychlorinated biphenyls in water samples. AB - A solid-phase microextraction (SPME) method has been developed for the quantification of polychlorinated biphenyls (PCBs) in water samples. Parameters such as sampling time, volume of water, volume of headspace, temperature, addition of salts, and agitation of the sample were studied. Because the time for reaching equilibrium between phases takes several hours or days, depending on the experimental conditions, it was necessary to work in nonequilibrium conditions to keep the sample analysis to a reasonable time. The possibility of sampling the headspace over the water sample (HSSPME), instead of immersing the fiber into the water (SPME), was also investigated, and despite the low partition of PCB into the headspace, HSSPME offered higher sensitivity than SPME at 100 degrees C. The adsorption kinetics for SPME at room temperature, SPME at 100 degrees C, and HSSPME at 100 degrees C were investigated and compared. The proposed HSSPME method exhibits excellent linearity and sensitivity. The detection limit was in the sub-ng/L level. This method has been applied to a real industrial harbor water and compared with liquid-liquid extraction. Both techniques offered similar results, but HSSPME was much more sensitive and considerably faster, by eliminating all the manual process intensive sample workup, and reduces solvent consumption entirely. The only drawback was that matrix effects were observed, but with the addition of deuterated surrogates to the sample or the use of a standard addition calibration, accurate quantification can be achieved. PMID- 21644773 TI - Shape-selective retention of polycyclic aromatic hydrocarbons on metalloprotoporphyrin-silica phases: effect of metal ion center and porphyrin coverage. AB - Various metalloprotoporphyrins (MProP) covalently linked to silica are examined as stationary phases for reversed-phase HPLC separation of polycyclic aromatic hydrocarbons (PAHs). The MProP-silica stationary phases are shown to exhibit extraordinary shape selectivity for planar over nonplanar PAHs, with the selectivity factors for the triphenylene/o-terphenyl solute pair approaching 30 on Cu(II)ProP-silica phases using 100% acetonitrile as the mobile phase. Shape selectivity and solute retention are highly dependent on the metal ion (M) within the center of the immobilized protoporphyrin (ProP) structure in accordance with the following sequence: Cu(II) > Fe(III) > Ni(II) > H(2) > Zn(II) ~ Cd(II). A face-to-face pi-pi interaction is believed to be the major retention mechanism of PAHs on ProP- and MProP-silica phases, with varying metal ion centers affecting the strength of this interaction. Beyond the influence of the central metal, varying surface coverages of metalloporphyrin also lead to significant changes in observed capacity factors and shape selectivities for PAH solutes. The extremely high shape selectivity for planar vs nonplanar PAHs suggests that MProP-silicas could be ideal materials for selectively preconcentrating the more toxic planar PAHs from environmental samples. Preliminary results relevant to this application are reported. PMID- 21644774 TI - Composite Films of Prussian Blue and N-Substituted Polypyrroles: Fabrication and Application to Optical Determination of pH. AB - A new and simple chemical method for deposition of thin blue films composed of Prussian Blue and N-substituted polypyrroles on nonconductive supports is presented. It is found that only pyrroles which are difficult to polymerize can be used for the preparation of such films. The resulting composite films were examined by SEM-EDAX, vis-NIR, and IR spectroscopy. The films are stable, thin, homogeneous, and optically transparent. The absorption maxima are at 720 nm, and spectral changes can be monitored using semiconductor light sources and detectors. The composite films are shown to be suitable for optical determination of pH over the pH 5-9 range because their absorbance strongly depends on pH in the physiological pH range. However, they undergo irreversible spectral changes if exposed to pH's >9. The films represent an alternative to indicator-based pH sensor materials because they do not require a dye to be immobilized. The pH measurements are highly reproducible, reversible in the physiological range, and not affected by ionic strength, alkaline cations, and typical oxidants and reductants. PMID- 21644775 TI - Automated Particle Analysis of Populations of Silver Halide Microcrystals by Electron Probe Microanalysis under Cryogenic Conditions. AB - An automated particle analysis routine is implemented on an electron microprobe for analyzing the chemical composition and projective area of populations of individual silver halide microcrystals. An LN2 cryostage is used to prevent material degradation due to reaction with the impinging electron beam. The background in the EDX spectra is lowered by depositing the microcrystals on a carbon-coated copper grid, mounted in a transmission holder. The ILalpha/AgLalpha net X-ray intensity ratio, obtained from a spectrum-fitting algorithm, is used to determine the crystal composition by means of a standard-based calibration curve. The uncertainty on the concentration measurement of individual microcrystals is calculated using the uncertainties on the net X-ray counts and the uncertainties on the calibration curve. The area measurement is optimized by introducing a gray value histogram correction on each individual measurement. Overlapping microcrystals are scrapped from the analysis by defining a maximum shape factor, against which the shape factor of the microcrystal is tested. To minimize problems with drift of the cryostage, spectrum acquisition is carried out immediately after a single microcrystal has been located, based on the backscattered electron image. Several applications are discussed. PMID- 21644776 TI - Total internal reflection fluorescence correlation spectroscopy for counting molecules at solid/liquid interfaces. AB - Fluorescence correlation spectroscopy, using tota internal reflection excitation (TIRFCS), is developed as a method to allow quantitative determination of molecular populations at solid/liquid interfaces. Population fluctuations of fluorescent molecules at the interface are observed as excess low-frequency noise on a fluorescence signal. Since the noise arises from molecular origins, its magnitude can be evaluated by Poisson statistics to determine the number of molecules in the interface volume. This quantitative information is available without sensitivity calibration or the preparation of standards and without fitting the transients to a kinetic model. Unlike single-molecule counting measurements, TIRFCS can produce these quantitative results even when the number of photoelectrons detected per molecule is small. Surface populations of rhodamine 6G dye molecules were measured at C-18-derivatized, flat silica surfaces in contact with aqueous solutions and compared with predicted values derived from chromatographic retention data. In addition, electrostatic and nonpolar contributions to the free energy of adsorption of the dye to C-18 modified silica surfaces were examined. PMID- 21644777 TI - Time-resolved fluorescence monitoring of aromatic radicals in photoinitiated processes. AB - Photolytic initiation of free radical reactions is important to many areas of technology; time-resolved monitoring of submicromolar concentrations of radicals produced during the course of these reactions is needed to provide information about the rate of initiation and its competition with radical recombination. In this work, time-resolved laser-induced fluorescence is evaluated for monitoring of diphenylketyl radicals produced by photoreduction of the triplet state of benzophenone. Fluorescence from the doublet-doublet transition of the radical is excited with a continuous wave laser and provides a sensitive method to detect these intermediates at nanomolar concentrations and to study their kinetics in solution on time scales from a few microseconds to hundreds of milliseconds. The ketyl radical fluorescence measurements of radical initiation reactions allowed the H atom abstraction rate constant by triplet benzophenone from both 2-propanol and benzhydrol to be determined, where k(H) = (2.1 +/- 0.1) * 10(6) M(-)(1) s( )(1) for 2-propanol and k(H) = (4.4 +/- 0.1) * 10(6) M(-)(1) s(-)(1) for benzhydrol. The diphenylketyl radical recombination rate constant was also determined by time-resolved fluorescence monitoring of the decay of the radical population and found to be k(r) = (1.9 +/- 0.2) * 10(8) M(-)(1) s(-)(1). Formation kinetics could be measured on a microsecond time scale from radical populations as low as 45 nM; decay kinetics could be followed on a millisecond time scale from 20 nM radical concentrations. PMID- 21644778 TI - Viscoelastic properties of low-viscosity liquids studied with thickness-shear mode resonators. AB - The network analysis method was applied to AT cut quartz blanks (f(0) = 10 MHz), which were loaded with liquids of low and medium viscosity (water, methanol, ethanol, 1-propanol, 1-butanol, glycerol solutions). The shift of the resonance frequency Deltaf could be separated into a term due to rigidly coupled mass Deltaf(rig) and a term due to viscous damping Deltaf - Deltaf(rig). From the difference Deltaf - Deltaf(rig) and the broadening of the resonance curve, the complex shear modulus G = G' + iomegaeta(L) was calculated. The viscosity coefficients eta(L) are in good agreement with literature data. As G' > 0, it can be concluded that the examined fluids also reveal elasticity at shear frequencies in the MHz range. For the low-viscosity liquids, elastic contributions resulting from collective interactions of molecules are measurable but small and neglectable in most applications. The medium viscous liquid glycerol (98%) begins to exhibit considerable elasticity, resulting from the relaxation of separate molecules. PMID- 21644779 TI - Characterization of lithium sulfate as an unsymmetrical-valence salt bridge for the minimization of liquid junction potentials in aqueous-organic solvent mixtures. AB - The transference numbers t of lithium sulfate in acetonitrile/water and methanol/water solvent mixtures have been studied by measuring the emfs of such transference cells as Pb(Hg)|PbSO(4)|Li(2)SO(4) (m(2))||Li(2)SO(4) (m(1))|PbSO(4)| Pb(Hg), Hg|Hg(2)SO(4)|Li(2)SO(4) (m(2))||Li(2)SO(4) (m(1))|Hg(2)SO(4)|Hg, and Li(x)Hg(1-)(x)|Li(2)SO(4) (m(1))||Li(2)SO(4) (m(2))|Li(x)Hg(1-)(x), in view of characterizing Li(2)SO(4) as an unsymmetrical salt bridge for the minimization of liquid junction potentials in potentiometric applications. In water, Li(2)SO(4) is nearly as good a salt bridge as the popular KCl one. Its effectiveness has been verified through the operational pH-metric cell using various aqueous pH(S) standards. In acetonitrile/water solvents of 0.09 mass fraction of acetonitrile at 298.15 K, Li(2)SO(4) shows exact ionic equitransference, obeying the general condition t(+)/z(+) = t(-)/|z(-)|. The Li(2)SO(4) salt bridge can be used either for simple insertion between two different electrolyte solutions to minimize the intervening liquid junction potentials, as the outer component of a double-bridge arrangement of any commonly available reference electrode, or, obviously, for structural incorporation in sulfate-reversible reference electrodes as the appropriate supporting and bridge electrolyte. The operational potential of the Hg| Hg(2)SO(4)|2 m Li(2)SO(4) reference electrode in aqueous solutions is 0.6326 V at 298.15 K. PMID- 21644780 TI - XPS Study of Counterion Incorporation into Poly(vinylferrocene). AB - XPS provides answers to two significant issues that arise in studying electroactive polymers. First, what fraction of possible charge centers participates in the redox process? Second, what are the relative partition coefficients of two counterions that compete as charge-compensating counterions? To answer both these questions, we emersed (or immersed) poly(vinylferrocene) films from (or into) supporting electrolytes while maintaining potential control to govern the electrochemical history of the electroactive film. We found that all ferrocene units in the surface of a poly(vinylferrocene) film deposited on gold can be oxidized when the counterion is perchlorate, tetrafluoroborate, hexafluorophosphate, tosylate, or 1,5-naphthalenedisulfonate. In answering the second question, no internal standard was needed: the concentration ratios of the two competing ions in both water and polymer phases were used as the experimental variables. For the completely oxidized ferricinium film, we found that the partition coefficient of perchlorate ions is 67% of that of fluoroborate ions. PMID- 21644781 TI - (13)C dynamic nuclear polarization: an alternative detector for recycled-flow NMR experiments. AB - Static NMR experiments on insensitive nuclei (e.g., (13)C) have been generally difficult because of low magnetogyric ratios and/or poor natural abundance, even at the highest magnetic fields available. One method of overcoming these sensitivity limitations is dynamic nuclear polarization (DNP), which can be utilized to enhance NMR signals relative to those obtained with static NMR detection. In this study, we report a recycled-flow DNP apparatus capable of monitoring (13)C scalar-dominated enhancements 1-2 orders of magnitude greater than those obtainable using the conventional recycled-flow (13)C NMR experiment. Specifically, a mixture of benzene and several chlorocarbons were continuously "recycled" through a (13)C DNP spectrometer. Results indicate successful detection with the (13)C DNP approach in scalar-dominated cases, where analytes could not be observed with conventional static or flow NMR detection in a reasonable length of time. PMID- 21644782 TI - Lowering the UV Absorbance Detection Limit in Capillary Zone Electrophoresis Using a Single Linear Photodiode Array Detector. AB - A new approach for lowering the UV absorbance detection limit in capillary electrophoresis is presented. This approach involves the use of a photodiode array in which each of the diodes in the array is treated as an independent detector. Over the course of a run, therefore, an electropherogram is generated for each diode in the array. Averaging the electropherograms generated from 1500 diodes in a diode array resulted in a signal-to-noise ratio 85 times that of an electropherogram generated from any one diode in the array. These signal-to-noise improvements are discussed, and the detection limits are compared to the detection limits obtained from a commercial single-point detector. The array detector improves the detection limit by a factor of 3.8 (+/-0.4). PMID- 21644783 TI - Simultaneous Determination of Volatile Metal (Pb, Hg, Sn, In, Ga) and Nonmetal Species (Se, P, As) in Different Atmospheres by Cryofocusing and Detection by ICPMS. AB - A sensitive method for multielemental speciation analysis of volatile metal and metalloid compounds in air has been developed. The analytes are sampled simultaneously in the field by cryofocusing on a small glass wool-packed column at -175 degrees C. Detection is performed in the laboratory by low-temperature GC hyphenated with ICPMS. Oxygen addition in the carrier gas was used to reduce interferences originating from the presence of volatile carbon-containing species in the samples. Plasma stability during analysis was monitored continuously by internal standardization (Xe). This system provides routine absolute detection limits of 0.06-0.07 pg (as Pb) for tetraalkyllead species (Me(4)Pb, Et(4)Pb), 0.2 pg (as Sn) for tetraalkyltin species (Me(4)Sn, Et(4)Sn), 0.8 pg (as Hg) for mercury species (Hg(0), Me(2)Hg, Et(2)Hg), and 2.5 pg (as Se) for selenium species (Me(2)Se). This instrumentation makes it possible to collect small air sample volumes and has been successfully applied to the determination of volatile metal and metalloid species in the atmosphere in urban and rural locations. Qualitative application in the semiconductor industry is also reported with regard to the detection of arsenic (ASH(3), tert-butylarsine), phosphorus (PH(3), tert-butylphosphine), alkylindium, and gallium species. PMID- 21644784 TI - Near-Field Surface-Enhanced Raman Imaging of Dye-Labeled DNA with 100-nm Resolution. AB - Raman chemical imaging on a scale of 100 nm is demonstrated for the first time. This is made possible by the combination of scanning near-field optical microscopy (SNOM or NSOM) and surface-enhanced Raman scattering (SERS), using brilliant cresyl blue (BCB)-labeled DNA as a sample. SERS substrates were produced by evaporating silver layers on Teflon nanospheres. The near-field SERS spectra were measured with an exposure time of 60 s and yielded good signal-to noise ratios (25:1). The distinction between reflected light from the excitation laser and Raman scattered light allows the local sample reflectivity to be separated from the signal of the adsorbed DNA molecules. This is of general importance to correct for topographic coupling that often occurs in near-field optical imaging. The presented data show a lateral dependence of the Raman signals that points to special surface sites with particularly high SERS enhancement. PMID- 21644785 TI - Photobleaching of Fluorescent Dyes under Conditions Used for Single-Molecule Detection: Evidence of Two-Step Photolysis. AB - The photostability of fluorescent dyes is of crucial importance for the statistical accuracy of single-molecule detection (SMD) and for the image quality of scanning confocal microscopy. Concurrent results for the photostability were obtained by two different experimental techniques. First, the photostabilities of several coumarin and rhodamine derivatives in aqueous solution were obtained by monitoring the steady-state fluorescence decay in a quartz cell. Furthermore, an epi-illuminated microscope, continuous wave (CW) excitation at 514.5 nm, and fluorescence correlation spectroscopy (FCS) with a newly developed theory were used to study the photobleaching characteristics of rhodamines under conditions used for SMD. Depending on the rhodamine structure, the probability of photobleaching, p(b), is in the order of 10(-)(6)-10(-)(7) for irradiances below 10(3) W/cm(2). However, a considerable increase of p(b) for irradiances above this level was observed which can only be described by photobleaching reactions from higher excited states (two-step photolysis). In view of these observations, the probability of photobleaching, p(b), as well as a closed expression of its dependence on the CW excitation irradiance considering a five-level molecular electronic state model with the possibility of photobleaching from higher excited electronic states, is derived. From this model, optimal conditions for SMD with respect to the number of emitted fluorescence photons and to the signal-to background ratio are discussed, taking into account both saturation and photobleaching. The additional photobleaching due to two-step photolysis limits the applicable irradiance. PMID- 21644786 TI - Laser mass spectrometric analysis of polycyclic aromatic hydrocarbons with wide wavelength range laser multiphoton ionization spectroscopy. AB - In many analytical techniques, 1+1 resonance-enhanced multiphoton ionization (1+1 REMPI) is used because it is an efficient and optically selective soft ionization method. While 1+1 REMPI of jet-cooled molecules has been extensively studied, little has been reported so far about this mechanism as it is used in analytical techniques, that is, in the cases where the molecules are not jet-cooled and where widely varying ionization wavelengths are employed. We used two-step laser mass spectrometry (L2MS) to study the wavelength (238-310 nm) dependence and the laser pulse energy dependence of the ion yield for 17 polycyclic aromatic hydrocarbons (PAHs). We discuss how these data allow prediction of the efficiency of 1+1 REMPI for a given compound. These advances open new perspectives for better understanding the L2MS spectra obtained directly from complex mixtures such as environmental samples. PMID- 21644787 TI - Use of triethylenetetraminehexaacetic Acid combined with field-amplified sample injection in speciation analysis by capillary electrophoresis. AB - Simultaneous speciation of lead, mercury, and selenium was carried out by capillary electrophoresis. The method used a polyaminocarboxylic acid, triethylenetetraminehexaacetic acid (TTHA), as an off-column complexing agent to form UV-absorbing complexes with the analytes for direct UV detection. TTHA was also added to the background electrolyte for the on-column complexations of the analytes, as well as for improving resolution and detection. To describe the migration behavior of the complexes, a theoretical model, considerating pH value and the concentrations of TTHA and SDS, was proposed. The parameters in the model were calculated on the basis of the experimental data, by nonlinear regression. The results were in good agreement with those from the literature. The model can be used for the prediction of migration behavior and for the optimization of the separation conditions. Field-amplified stacking injection was performed because the complexes were charged. Up to 1500-fold on-line enrichment and down to sub nanogram-per-milliliter detection limits were obtained for the analytes under the optimal stacking conditions. Finally, the applicability of the method was evaluated on seawater samples. PMID- 21644788 TI - Determination of aqueous solubility and surface adsorption of polycyclic aromatic hydrocarbons by laser multiphoton ionization. AB - Polycyclic aromatic hydrocarbons (PAHs) are of considerable analytical interest due to their environmental effects. On-line monitoring of these compounds based on microdroplet sampling requires accurate characterization of their solubilities and surface adsorption. A new method to determine aqueous solubility and surface adsorption of PAHs is suggested, using a combination of microdroplet sampling and multiphoton ionization-based fast-conductivity (MPI-FC) techniques. Controlled droplet contamination by PAHs was performed by external deposition upon direct contact with renewable water droplets (?10 MUL). Our approach relies on the finding that, at the onset of aqueous equilibrium solubility, X(E), a sharp increase in the detected photocharges is recorded. We show that this is directly related to enhanced surface adsorption of a particular PAH material. This behavior manifests itself in slope variation of the respective calibration curves. The PAH materials used, which have provided about four decades variation in X(E) figures, were anthracene, perylene, pyrene, and phenanthrene. Applicability of the method for studying the surface excess and adsorption is specified. The MPI-FC technique provides an easy analytical tool for measuring low aqueous solubility of organic compounds and characterization of their adsorption, with no need for laborious sample preparation routines. PMID- 21644789 TI - Mass spectrometry of particles formed in a deuterated ethene diffusion flame. AB - Nanometer-sized spherule soot precursor particles have been collected by thermophoretic sampling from the interior of a laminar diffusion flame and mass analyzed by laser microprobe mass spectrometry. Mass spectra of the precursor particles formed in an ethene diffusion flame have indicated the presence of polycyclic aromatic hydrocarbons (PAHs) in the m/z range of 202-300 and higher mass peaks extending out to m/z 472. The mass resolution of the time-of-flight mass spectrometer used did not provide conclusive identification of PAHs because of ambiguities in assignment for the relative amounts of carbon and hydrogen (C(x)H(y)) for each PAH peak and the possibilities of spectral interferences. To determine the chemical formula that can be assigned to each molecular ion peak, an isotopically pure deuterated ethene (C(2)D(4)) fuel was burned in place of normal ethene (C(2)H(4)) in the diffusion flame. For the normal ethene fuel, mass peaks tentatively identified as C(16)H(10) to C(38)H(16) were obtained. Accordingly, deuterated PAH peaks ranging from C(16)D(10) to C(38)D(16) were found when C(2)D(4) was burned. These m/z values correspond to molecular ion, M*(+), peaks for an array of PAH compounds. The deuterated PAH mass peaks (C(x)D(y)) were entirely consistent with a mass shift of y mass units with respect to the normal PAH mass peaks. The carbonaceous particle aggregates collected from the upper flame region have mass peaks characteristic of C(x)(+) and C(x)H(+), while the deuterated soot has C(x)(+) and C(x)D(+). The deuterated ethene experiment has verified the identities of x and y in the PAH (C(x)H(y)) compounds present in the precursor particle samples. No prior experiment using pure deuterium-based fuel as a combustion diagnostic to form aerosol-containing deuterated PAH compounds has been reported. PMID- 21644790 TI - On-Line Analysis of Stable Isotopes of Nitrogen in NH(3), NO, and NO(2) at Natural Abundance Levels. AB - Methods were developed for the on-line analysis of stable isotopes of nitrogen (at natural abundance levels) in NH(3), NO and NO(2) in order to study the contribution of these trace gases to nitrogen cycling in ecosystems. Standard methods for the combustion of organic substances to N(2) (for the determination of the (15)N/(14)N ratios by mass spectrometry) failed to satisfactorily oxidize or reduce the respective trace gases. The following oxidation or reduction techniques ensured quantitative conversion of the trace gases to N(2) and analytical precision close to the internal precision of the instrument (precision +/-0.150/00): (1) oxidation of NH(3) to N(2) at 1150 degrees C on a NiO surface (which needs a reoxidation before each NH(3) analysis; measuring range 11.7-58.8 nmol of NH(3), precision +/-0.300/00); (2) reduction of NO to N(2) at 1150 degrees C on a Ni surface (measuring range 33.3-133.3 nmol of NO, precision +/ 0.280/00); (3) reduction of NO(2) to form NO at 420 degrees C on a Mo surface followed by further treatment as for NO (measuring range 26.0-129.9 nmol of NO(2), precision +/-0.900/00). This last technique was developed due to the poor chromatographic properties of NO(2). PMID- 21644791 TI - Gas chromatography connected to multiple channel electrospray ionization mass spectrometry for the detection of volatile organic compounds. AB - This work successfully connected gas chromatography (GC) to seven-channel electrospray ionization (ESI) mass spectrometry to separate and detect a mixture of volatile organic compounds. Gaseous analyte was eluted separately from a GC column and directed into the central channel of the ESI source. The analyte was protonated by ion-molecule reactions between the analyte and the ions which were generated by electrospraying the acidic solution through the outside six channels surrounding the central channel. Real-time analysis of the organic reaction involving volatile and thermally unstable compounds (dimethylhydrazine ? azomethane + H(2)) was also achieved by continuously purging the air in the reaction vessel to the seven-channel ESI source. PMID- 21644792 TI - Quantitative determination of bucindolol concentration in intact gel capsules using Raman spectroscopy. AB - The ideal quality control method for pharmaceutical products should be capable of rapid nondestructive testing of intact tablets or capsules. Raman spectroscopy using near-infrared excitation is shown to be capable of obtaining useful spectral data directly from drug formulations in gel capsules and from the gel capsules inside blister packs. The Raman data collected from the capsules inside blister packs containing 0-100 mg of the active ingredient (bucindolol), when coupled with multivariate calibration, resulted in a calibration SEP of 3.36 mg. The largest source of error was found to be due to sample inhomogeneity. Even so, the method is shown to have significant potential as a rapid nondestructive quality control method for pharmaceutical samples. PMID- 21644793 TI - Investigation of Enantiomer Bonding on a Chiral Stationary Phase by FT-Raman Spectrometry. AB - The bonding of serine, phenylalanine, and mandelic acid enantiomers on an N-3,5 dinitrobenzoyl-l-leucine chiral stationary phase (on zeolite A support) has been investigated by FT-Raman spectrometry. It was found that retention is due to hydrogen bonds and pi-stacking interactions between the stationary phase and the analyte. The involvement of the two different amide groups (as donor and/or acceptor) in the complexation reaction can be followed based on spectral data. A correlation was found between the ratio of the amide I and the ring stretching (1532 cm(-)(1)) bands and retention data. PMID- 21644794 TI - Fluorescent Functional Recognition Sites through Molecular Imprinting. A Polymer Based Fluorescent Chemosensor for Aqueous cAMP. PMID- 21644795 TI - Water-soluble iron oxide nanoparticles with high stability and selective surface functionality. AB - The water dispensability and stability of high quality iron oxide nanoparticles synthesized in organic solvents are major issues for biomedical and biological applications. In this paper, a versatile approach for preparing water-soluble iron oxide nanoparticles with great stability and selective surface functionality (-COOH, -NH(2), or -SH) was demonstrated. The hydrophobic nanoparticles were first synthesized by the thermal decomposition of an iron oleate complex in organic solvent. Subsequently, the hydrophobic coatings of nanoparticles were replaced with poly(acrylic acid) , polyethylenimine, or glutathione, yielding charged nanoparticles in aqueous solution. Two parameters were found to be critical for obtaining highly stable nanoparticle dispersions: the original coating and the surfactant-to-nanoparticle ratio. These charged nanoparticles exhibited different stabilities in biological buffers, which were directly influenced by the surface coatings. This report will provide significant practical value in exploring the biological or biomedical applications of iron oxide nanoparticles. PMID- 21644796 TI - In vitro metabonomic study detects increases in UDP-GlcNAc and UDP-GalNAc, as early phase markers of cisplatin treatment response in brain tumor cells. AB - O-linked beta-N-acetylglucosamine glycosylation (O-GlcNAcylation) is important in a number of biological processes and diseases including transcription, cell stress, diabetes, and neurodegeneration and may be a marker of tumor metastasis. Uridine diphospho-N-acetylglucosamine (UDP-GlcNAc), the donor molecule in O GlcNAcylation, can be detected by (1)H nuclear magnetic resonance spectroscopy ((1)H NMR), giving the potential to measure its level noninvasively, providing a novel biomarker of prognosis and treatment monitoring. In this in vitro metabonomic study, four brain cancer cell lines were exposed to cisplatin and studied for metabolic responses using (1)H NMR. The Alamar blue assay and DAPI staining were used to assess cell sensitivity to cisplatin treatment and to confirm cell death. It is shown that in the cisplatin responding cells, UDP GlcNAc and uridine diphospho-N-acetylgalactosamine (UDP-GalNAc), in parallel with (1)H NMR detected lipids, increased with cisplatin exposure before or at the onset of the microscopic signs of evolving cell death. The changes in UDP-GlcNAc and UDP-GalNAc were not detected in the nonresponders. These glycosylated UDP compounds, the key substrates for glycosylation of proteins and lipids, are commonly implicated in cancer proliferation and malignant transformation. However, the present study mechanistically links UDP-GlcNAc and UDP-GalNAc to cancer cell death following chemotherapeutic treatment. PMID- 21644797 TI - Gastroprotective and ulcer-healing mechanisms of ellagic acid in experimental rats. AB - Ellagic acid (EA), a plant-derived polyphenol, exhibits antioxidant, anti inflammatory, and gastroprotective effects. Its gastroprotective mechanisms have not been fully elucidated nor have its effects on chronic ulcer previously been described. Toward these ends, the antiulcer activities of EA were evaluated in acute (ethanol and indomethacin) and chronic (acetic acid) ulcer models in Wistar rats. In this study, oral administration of EA significantly prevented the gastric ulceration caused by ethanol, indomethacin, and acetic acid treatments. Its gastroprotective mechanism in ethanol-induced ulcer were partly due to intensification in the endogenous production of nitric oxide, an antioxidant effect by replenishing depletion of endogenous nonprotein sulfhydryls and attenuation of tumor necrosis factor-alpha increase, whereas in indomethacin ulcer, it is partly due to a reduction in the plasma level of leukotriene B(4). In acetic acid ulcer, promotion of ulcer-healing effects was partly due to attenuation of the elevated levels of the inflammatory cytokines TNF-alpha, interferon-gamma, and interleukins-4 and -6. These findings suggest that ellagic acid exerts its antiulcer activity by strengthening the defensive factors and attenuating the offensive factors. PMID- 21644798 TI - Quasiclassical trajectory study of the C(1D) + H2 reaction and isotopomeric variants: kinetic isotope effect and CD/CH branching ratio. AB - The recently proposed ab initio single-sheeted double many-body expansion potential energy for the methylene molecule has been used to perform quasiclassical trajectory (QCT) calculations for the title reaction. Thermal and initial state-specific (v = 0, j = 0) rate constants for the C((1)D) + H(2)/HD/D(2) reactions have been obtained over a wide range of temperatures. Cross sections for the reaction C((1)D) + H(2) and its deuterated isotopes have also been calculated, as well as the CD/CH branching ratios for the C((1)D) + HD reaction. It is found that the CD + H product channel in the C((1)D) + HD reaction is preferred relative to the CH + D channel. The estimated rate constants are predicted to be in the order k(H2) > k(HD) > k(D2) and the calculated cross sections and rate constants compared with available theoretical and experimental data. PMID- 21644799 TI - Glycyrrhizic acid and 18beta-glycyrrhetinic acid modulate lipopolysaccharide induced inflammatory response by suppression of NF-kappaB through PI3K p110delta and p110gamma inhibitions. AB - The roots and rhizomes of licorice ( Glycyrrhia ) species have been used extensively as natural sweeteners and herbal medicines. The aim of this work was to determine the in vitro anti-inflammatory effects of glycyrrhizic acid (GA) and 18beta-glycyrrhetinic acid (18betaGA) from licorice in a lipopolysaccharide (LPS) stimulated macrophage model. The results showed that treatment with 25-75 MUM GA or 18betaGA did not reduce RAW 264.7 cell viability but did significantly inhibit the production of LPS-induced nitric oxide (NO), prostaglandin E(2) (PGE(2)), and intracellular reactive oxygen species (ROS). Western blotting and reverse transcriptase polymerase chain reaction (RT-PCR) analyses revealed that GA and 18betaGA significantly reduced the protein and mRNA levels of iNOS and COX-2 in LPS-induced macrophages. Both GA and 18betaGA inhibited the activation of NF kappaB and the activities of phosphoinositide-3-kinase (PI3K) p110delta and p110gamma isoforms and then reduced the production of LPS-induced tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-6, and IL-1beta in a dose-dependent manner. In conclusion, these results indicate that GA and 18betaGA may provide an anti-inflammatory effect by attenuating the generation of excessive NO, PGE(2), and ROS and by suppressing the expression of pro-inflammatory genes through the inhibition of NF-kappaB and PI3K activity. Thus, the results suggest that GA and 18betaGA might serve as potential agents for the treatment of inflammatory mediated diseases. PMID- 21644801 TI - Are symptoms of ADHD related to substance use among college students? AB - Attention-deficit/hyperactivity disorder (ADHD) is a common childhood disorder that often persists into adolescence and adulthood and has been associated with an increased risk for substance use. Due to improved treatment and educational policies, more students with high levels of ADHD symptoms are attending college despite continued ADHD symptoms. Little research has examined whether college students with higher levels of ADHD symptoms are at increased risk for heavy substance use compared to college students with few ADHD symptoms. The current study examined the relation of ADHD symptoms to substance use (e.g., cigarette smoking, alcohol use, marijuana use, and cocaine use). We hypothesized that greater ADHD symptomatology (inattentive, hyperactive/impulsive, and total ADHD) would be related to higher rates of cigarette smoking, alcohol use, alcohol problems, and illicit drug use. Participants were 889 college undergraduates who completed an online survey. Results suggest that ADHD symptoms, particularly inattentive symptoms, were positively associated with cigarette smoking and problems associated with alcohol after controlling for conduct disorder (CD) symptoms. ADHD symptoms were not significantly associated with alcohol use or illicit drug use after the effects of CD symptoms were accounted for. Results have important implications for prevention and treatment of college student substance use. Limitations and future directions are discussed. PMID- 21644800 TI - Mechanistic strategies in the HDV ribozyme: chelated and diffuse metal ion interactions and active site protonation. AB - The crystal structure of the precleaved form of the hepatitis delta virus (HDV) ribozyme reveals two G*U wobbles near the active site: a rare reverse G*U wobble involving a syn G base, and a standard G*U wobble at the cleavage site. The catalytic mechanism for this ribozyme has been proposed to involve a Mg(2+) ion bound to the reverse G*U wobble, as well as a protonated C75 base. We carried out molecular dynamics simulations to analyze metal ion interaction with the reverse and standard G*U wobbles and to investigate the impact of C75 protonation on the structure and motions of the ribozyme. We identified two types of Mg(2+) ions associated with the ribozyme, chelated and diffuse, at the reverse and standard G*U wobbles, respectively, which appear to contribute to catalysis and stability, respectively. These two metal ion sites exhibit relatively independent behavior. Protonation of C75 was observed to locally organize the active site in a manner that facilitates the catalytic mechanism, in which C75(+) acts as a general acid and Mg(2+) as a Lewis acid. The simulations also indicated that the overall structure and thermal motions of the ribozyme are not significantly influenced by the catalytic Mg(2+) interaction or C75 protonation. This analysis suggests that the reaction pathway of the ribozyme is dominated by small local motions at the active site rather than large-scale global conformational changes. These results are consistent with a wealth of experimental data. PMID- 21644802 TI - Utilizing Social Action Theory as a framework to determine correlates of illicit drug use among young men who have sex with men. AB - Young men who have sex with men (YMSM) continue to be at elevated risk for substance use; however, models explaining this phenomenon have often focused on a limited array of explanatory constructs. This study utilizes Social Action Theory (SAT) as a framework to address gaps in research by documenting the social, behavioral, and demographic risk factors associated with illicit drug use among YMSM. Structural equation modeling was used to apply SAT to a cross-sectional sample of 526 men from the Healthy Young Men Study, a longitudinal study of substance use and sexual risk behavior among YMSM in Los Angeles. The final model possessed very good fit statistics (Comparative Fit Index (CFI) = 0.936, Tucker Lewis Index (TLI) = 0.925, Root Mean Square Error of Approximation (RMSEA) = 0.040) indicating that SAT is appropriate for use with YMSM. Substance use interventions for YMSM could be enhanced by employing SAT as conceptualized in this study and using a multitargeted strategy for impacting illicit drug use. PMID- 21644803 TI - Adolescent alcohol-related risk cognitions: the roles of social norms and social networking sites. AB - The present study examined the impact of socially based descriptive norms on willingness to drink alcohol, drinker prototype favorability, affective alcohol attitudes, and perceived vulnerability for alcohol-related consequences within the Prototype Willingness model. Descriptive norms were manipulated by having 189 young adolescents view experimenter-created profile pages from the social networking site Facebook, which either showed older peers drinking or not. The results provided evidence that descriptive norms for alcohol use, as portrayed by Facebook profiles, significantly impact willingness to use, prototypes, attitudes toward use, and perceived vulnerability. A multiple mediation analysis indicated that prototypes, attitudes, and perceptions of use mediated the relationship between the content of the Facebook profile and willingness. These results indicate that adolescents who perceive that alcohol use is normative, as evidenced by Facebook profiles, are at higher risk for cognitions shown to predict alcohol use than adolescents who do not see alcohol use portrayed as frequently on Facebook. PMID- 21644804 TI - What do women want? Alcohol treatment choices, treatment entry and retention. AB - Study aims were to assess preferences for individual or conjoint treatment, differences between women with alcohol use disorders (AUDs) selecting each modality, and the impact on treatment entry and retention of providing a choice of treatments. During initial screening, women with AUDs in an intimate relationship with a male partner were given the choice of individual or conjoint treatment. After choosing a treatment modality and completing all assessments they entered one of two randomized trials testing different approaches to each treatment modality. Standardized measures were used to assess drinking quantity, frequency, and problems; motivation to change; and relationship satisfaction. Women's reasons for choice of treatment modality were coded using an iterative coding process. Results showed that women were more likely to select and follow through with individual than conjoint treatment. Women cited a desire to work on individual problems, lack of perceived support from their partner, and logistical issues as reasons for preferring individual treatment. Women in the two choice groups did not differ significantly on individual, partner, or relationship characteristics, but small to medium effect sizes suggested that women choosing individual treatment were more educated and less satisfied with their relationship, had fewer pretreatment heavy drinking days, and heavier drinking partners. Offering women a choice of treatment modality increased the probability of entering treatment, but not treatment attendance. Results suggest that barriers to couple therapy for women with AUDs need to be addressed to facilitate more widespread dissemination. Given women's preferences, it also is important to offer a range of treatments. PMID- 21644805 TI - Prospective evaluation of the effects of anxiety sensitivity and state anxiety in predicting acute nicotine withdrawal symptoms during smoking cessation. AB - The current investigation explored the main and interactive effects of anxiety sensitivity (AS) and state anxiety in predicting acute nicotine withdrawal symptoms experienced during the initial 14 days of smoking cessation. Participants included 123 adult daily smokers (84 women; Mage = 45.93 years, SD = 10.34) undergoing psychosocial-pharmacological cessation treatment. Results indicated that after controlling for the effects of participant sex and nicotine dependence, state anxiety but not AS significantly predicted initial levels of nicotine withdrawal symptoms. Results also demonstrated that both state anxiety and AS were significantly related to the change in nicotine withdrawal symptoms over time. Finally, our results revealed a significant interaction between AS and state anxiety. Specifically, higher levels of AS were associated with a stronger relation between state anxiety and nicotine withdrawal symptoms experienced during the cessation attempt. Results suggest that among high AS persons, state anxiety may be more relevant, compared to those low in AS, in regard to experiencing withdrawal symptoms as more intense during the early phases of quitting. PMID- 21644807 TI - Anticipated regret and organ donor registration--a pilot study. AB - OBJECTIVE: In this pilot study we tested the influence of manipulating anticipated regret on organ donor registration behavior. METHOD: A simple web based experimental trial was conducted. Nonorgan donors were allocated to a simple anticipated regret manipulation versus a theory of planned behavior or a control condition. The main outcome measure was registration on the U.K. organ donor register at 1-month follow-up. RESULTS: A simple anticipated regret manipulation led to a significant increase in organ donor registrations. CONCLUSION: Interventions utilizing anticipated regret may have the potential to significantly increase organ donation rates. PMID- 21644806 TI - Implicit measures of early-life family conditions: relationships to psychosocial characteristics and cardiovascular disease risk in adulthood. AB - OBJECTIVES: An implicit measure of early-life family conditions was created to help address potential biases in responses to self-reported questionnaires of early-life family environments. We investigated whether a computerized affect attribution paradigm designed to capture implicit, affective responses (anger, fear, warmth) regarding early-life family environments was (a) stable over time, (b) associated with self-reports of childhood family environments, (c) able to predict adult psychosocial profiles (perceived social support, heightened vigilance), and (d) able to predict adult cardiovascular risk (blood pressure) either alone or in conjunction with a measure of early-life socioeconomic status. METHOD: Two studies were conducted to examine reliability and validity of the affect attribution paradigm (Study 1, N = 94) and associated adult psychosocial outcomes and cardiovascular risk (Study 2, N = 122). RESULTS: Responses on the affect attribution paradigm showed significant correlations over a 6-month period, and were moderately associated with self-reports of childhood family environments. Greater attributed negative affect about early-life family conditions predicted lower levels of current perceived social support and heightened vigilance in adulthood. Attributed negative affect also interacted with early-life socioeconomic status (SES) to marginally predict resting systolic blood pressure (SBP), such that those individuals high in early-life SES but who had implicit negative affect attributed to early-life family conditions had SBP levels that were as high as individuals low in early-life SES. CONCLUSION: Implicit measures of early-life family conditions are a useful approach for assessing the psychosocial nature of early-life environments and linking them to adult psychosocial and physiological health profiles. PMID- 21644808 TI - Reverse causation in activity-cognitive ability associations: the Lothian Birth Cohort 1936. AB - Active lifestyles might protect cognitive abilities; however, studies rarely consider the reverse causal direction. Activity-cognition associations might reflect stable intelligence differences rather than a protective effect of activity. The Lothian Birth Cohort 1936 (n = 1091) completed cognitive tests aged 70, having taken an intelligence test aged 11. Activity (assessed by participation in 15 activities that produced a socio-intellectual activity factor, and by physical activity) was positively associated with cognition (r = .08 to .32, p <= .05). When age-11 IQ and adult social class were controlled, only physical activity remained significantly associated with general cognitive ability and processing speed. PMID- 21644809 TI - Unconscious vigilance: worldview defense without adaptations for terror, coalition, or uncertainty management. AB - Individuals subtly reminded of death, coalitional challenges, or feelings of uncertainty display exaggerated preferences for affirmations and against criticisms of their cultural in-groups. Terror management, coalitional psychology, and uncertainty management theories postulate this "worldview defense" effect as the output of mechanisms evolved either to allay the fear of death, foster social support, or reduce anxiety by increasing adherence to cultural values. In 4 studies, we report evidence for an alternative perspective. We argue that worldview defense owes to unconscious vigilance, a state of accentuated reactivity to affective targets (which need not relate to cultural worldviews) that follows detection of subtle alarm cues (which need not pertain to death, coalitional challenges, or uncertainty). In Studies 1 and 2, death primed participants produced exaggerated ratings of worldview-neutral affective targets. In Studies 3 and 4, subliminal threat manipulations unrelated to death, coalitional challenges, or uncertainty evoked worldview defense. These results are discussed as they inform evolutionary interpretations of worldview defense and future investigations of the influence of unconscious alarm on judgment. (PsycINFO Database Record (c) 2011 APA, all rights reserved). PMID- 21644811 TI - Post-concussion cognitive declines and symptomatology are not related to concussion biomechanics in high school football players. AB - Concussion is a major public health concern with nearly 4 million injuries occurring each year in the United States. In the acute post-injury stage, concussed individuals demonstrate cognitive function and motor control declines as well as reporting increased symptoms. Researchers have hypothesized that the severity of these impairments is related to impact magnitude. Using the Head Impact Telemetry System (HITS) to record head impact biomechanics, we sought to correlate pre- and post-concussive impact characteristics with declines in cognitive performance and increases in concussion-related symptoms. Over four seasons, 19 high school football athletes wearing instrumented helmets sustained 20 diagnosed concussions. Each athlete completed a baseline computer-based symptom and cognitive assessment during the pre-season and a post-injury assessment within 24 h of injury. Correlational analyses identified no significant relationships between symptoms and cognitive performance change scores and impact biomechanics (i.e., time from session start until injury, time from the previous impact, peak linear acceleration, peak rotational acceleration, and HIT severity profile [HITsp]). Nor were there any significant relationships between change scores and the number of impacts, cumulative linear acceleration, cumulative rotational acceleration, or cumulative HITsp values associated with all impacts prior to or following the injury. This investigation is the first to examine the relationship between concussion impact characteristics, including cumulative impact profiles, and post-morbid outcomes in high school athletes. There appears to be no association between head impact biomechanics and post concussive outcomes. As such, the use of biomechanical variables to predict injury severity does not appear feasible at this time. PMID- 21644810 TI - Recommendations for the use of common outcome measures in pediatric traumatic brain injury research. AB - This article addresses the need for age-relevant outcome measures for traumatic brain injury (TBI) research and summarizes the recommendations by the inter agency Pediatric TBI Outcomes Workgroup. The Pediatric Workgroup's recommendations address primary clinical research objectives including characterizing course of recovery from TBI, prediction of later outcome, measurement of treatment effects, and comparison of outcomes across studies. Consistent with other Common Data Elements (CDE) Workgroups, the Pediatric TBI Outcomes Workgroup adopted the standard three-tier system in its selection of measures. In the first tier, core measures included valid, robust, and widely applicable outcome measures with proven utility in pediatric TBI from each identified domain including academics, adaptive and daily living skills, family and environment, global outcome, health-related quality of life, infant and toddler measures, language and communication, neuropsychological impairment, physical functioning, psychiatric and psychological functioning, recovery of consciousness, social role participation and social competence, social cognition, and TBI-related symptoms. In the second tier, supplemental measures were recommended for consideration in TBI research focusing on specific topics or populations. In the third tier, emerging measures included important instruments currently under development, in the process of validation, or nearing the point of published findings that have significant potential to be superior to measures in the core and supplemental lists and may eventually replace them as evidence for their utility emerges. PMID- 21644813 TI - Traumatic brain injury in adult rats causes progressive nigrostriatal dopaminergic cell loss and enhanced vulnerability to the pesticide paraquat. AB - Parkinson's disease (PD) is a neurodegenerative disorder characterized by the loss of nigrostriatal dopaminergic neurons and the accumulation of alpha synuclein. Both traumatic brain injury (TBI) and pesticides are risk factors for PD, but whether TBI causes nigrostriatal dopaminergic cell loss in experimental models and whether it acts synergistically with pesticides is unknown. We have examined the acute and long-term effects of TBI and exposure to low doses of the pesticide paraquat, separately and in combination, on nigrostriatal dopaminergic neurons in adult male rats. In an acute study, rats received moderate TBI by lateral fluid percussion (LFP) injury, were injected with saline or paraquat (10 mg/kg IP) 3 and 6 days after LFP, were sacrificed 5 days later, and their brains processed for immunohistochemistry. TBI alone increased microglial activation in the substantia nigra, and caused a 15% loss of dopaminergic neurons ipsilaterally. Paraquat increased the TBI effect, causing a 30% bilateral loss of dopaminergic neurons, reduced striatal tyrosine hydroxylase (TH) immunoreactivity more than TBI alone, and induced alpha-synuclein accumulation in the substantia nigra pars compacta. In a long-term study, rats received moderate LFP, were injected with saline or paraquat at 21 and 22 weeks post-injury, and were sacrificed 4 weeks later. At 26 weeks post injury, TBI alone induced a 30% bilateral loss of dopaminergic neurons that was not exacerbated by paraquat. These data suggest that TBI is sufficient to induce a progressive degeneration of nigrostriatal dopaminergic neurons. Furthermore, TBI and pesticide exposure, when occurring within a defined time frame, could combine to increase the PD risk. PMID- 21644814 TI - Severity profile of penetrating ballistic-like brain injury on neurofunctional outcome, blood-brain barrier permeability, and brain edema formation. AB - This study evaluated the injury severity profile of unilateral, frontal penetrating ballistic-like brain injury (PBBI) on neurofunctional outcome, blood brain barrier (BBB) permeability, and brain edema formation. The degree of injury severity was determined by the delivery of a water-pressure pulse designed to produce a temporary cavity by rapid (<40 ms) expansion of the probe's elastic balloon calibrated to equal 5%, 10%, 12.5%, or 15% of total rat brain volume (control groups consisted of sham surgery or insertion of the probe only). Neurofunctional assessments revealed motor and cognitive deficits related to the degree of injury severity, with the most clear-cut profile of PBBI injury severity depicted by the Morris water maze (MWM) results. A biphasic pattern of BBB leakage was detected in the injured hemisphere at all injury severity levels at 4 h post-injury, and again at 48-72 h post-injury, which remained evident out to 7 days post-PBBI in the 10% and 12.5% PBBI groups. Likewise, significant brain edema was detected in the injured hemisphere by 4 h post-injury and remained elevated out to 7 days post-injury in the 10% and 12.5% PBBI groups. However, following 5% PBBI, significant levels of edema were only detected from 24 h to 48h post-injury. These results identify an injury severity profile of BBB permeability, brain edema, and neurofunctional impairment that provides sensitive and clinically relevant outcome metrics for studying potential therapeutics. PMID- 21644815 TI - Disconnecting the yin and yang relation of epidermal growth factor receptor (EGFR)-mediated delivery: a fully synthetic, EGFR-targeted gene transfer system avoiding receptor activation. AB - The epidermal growth factor receptor (EGFR) is upregulated within a high percentage of solid tumors and hence is an attractive target for tumor-targeted therapies including gene therapy. The natural EGFR ligand epidermal growth factor (EGF) has been used for this purpose, despite the risk of mitogenic effects due to EGFR activation. We have developed a fully synthetic, EGFR-targeted gene delivery system based on PEGylated linear polyethylenimine (LPEI), allowing evaluation of different EGFR-binding peptides in terms of transfection efficiency and EGFR activation. Peptide sequences directly derived from the human EGF molecule enhanced transfection efficiency with concomitant EGFR activation. Only the EGFR-binding peptide GE11, which has been identified by phage display technique, showed specific enhancement of transfection on EGFR-overexpressing tumor cells including glioblastoma and hepatoma, but without EGFR activation. EGFR targeting led to high levels of cell association of fluorescently labeled polyplexes after only 30 min of incubation. EGF pretreatment of cells induced enhanced cellular internalization of all polyplex types tested, pointing at generally enhanced macropinocytosis. EGF polyplexes diminished cell surface expression of EGFR for up to 4 hr, whereas GE11 polyplexes did not. In a clinically relevant orthotopic prostate cancer model, intratumorally injected GE11 polyplexes were superior in inducing transgene expression when compared with untargeted polyplexes. PMID- 21644817 TI - Factors associated with the development of severe asthma exacerbations in children. AB - BACKGROUND: Asthma exacerbations are one of the most common causes of hospitalization in children and account for approximately 10,000 intensive care unit (ICU) admissions per year in the United States. Despite the prevalence of this disease in children, the factors associated with the development of these severe exacerbations are largely unknown. METHODS: A retrospective case-control study was conducted involving all eligible children admitted to the hospital with asthma for a 1-year period. Potential associated factors and outcomes of children admitted to the ICU with a severe exacerbation (cases) were compared to those of children with acute asthma admitted to the ward (controls). RESULTS: A total of 188 children were hospitalized with asthma during the study period, 57 (30%) of whom required admission to the ICU. There were no differences in age, gender, or race between cases and controls. Children admitted to the ICU were significantly more likely to have an allergy or irritant-triggered exacerbation than children admitted to the ward (OR 3.9; 95% CI 1.9-8.2; p = .0003). Additionally, children in the ICU had a significantly shorter duration of illness before being admitted to the hospital compared to those admitted to the ward (1.7 +/- 2.3 vs. 3.4 +/- 4.8 days; p = .002). CONCLUSIONS: In this retrospective review, severe asthma exacerbations in children are associated with a more rapid onset of symptoms and are more likely to be associated with allergens or irritants, supporting the importance of atopy in this population. PMID- 21644818 TI - Proinflammatory effects of bare and PEGylated ORMOSIL-, PLGA- and SUV-NPs on monocytes and PMNs and their modulation by f-MLP. AB - AIMS: We wanted to test the proinflammatory effects of vinyltriethoxysilane-based organically modified silica nanoparticles (ORMOSIL-NPs) in vitro on blood leukocytes. MATERIALS & METHODS: Cell selectivity, cytokines/chemokines and O(2) (-) production were analyzed using nonpolyethylene glycol (PEG)ylated and PEGylated ORMOSIL-NPs, poly(lactic-co-glycolic acid) (PLGA)-NPs and small unilamellar vesicles (SUV)-NPs. RESULTS: ORMOSIL-NPs mostly bound to monocytes while other NPs to all leukocyte types similarly. Cell capture of PEGylated-NPs decreased strongly (ORMOSIL), moderately (PLGA) and weakly (SUV). Bare ORMOSIL NPs effectively stimulated the production of IL-1beta/IL-6/TNF-alpha/IL-8 by monocytes and of IL-8 by polymorphonuclear leukocytes (PMNs). NP PEGylation inhibited such effects only partially. Formyl-methionine-leucine phenylalanine (f MLP) further increased the release of cytokines/chemokines by monocytes/PMNs primed with bare and PEGylated ORMOSIL-NPs. PEGylated SUV-NPs, bare and PEGylated ORMOSIL- and PLGA-NPs sensitize PMNs and monocytes to secrete O(2) (-) upon f-MLP stimulation. CONCLUSION: ORMOSIL-NPs are preferentially captured by circulating monocytes but stimulate both monocytes and PMNs per se or by sensitizing them to another agonist (f-MLP). PEG-coating confers stealth effects but does not completely eliminate leukocyte activation. Safe nanomedical applications require the evaluation of both intrinsic and cooperative proinflammatory potential of NPs. PMID- 21644819 TI - Localization of matrix metalloproteinase (MMP)-9 in lung tissue of a murine model of allergic asthma. AB - MMP-9 (gelatinase B) is recognized in chronic obstructive pulmonary disease (COPD) and now asthma as playing a central role in matrix degradation in injury, as well as contributing to the remodeling process. The increasing focus on MMP-9 in human and animal research supports the need for a reliable immunostain in lung tissue. However, MMP-9 immunostaining in murine systems is hampered by several factors. First, many of the anti-human antibodies do not readily cross-react with murine MMP-9 despite the high degree of conservation between human and murine MMP 9. Secondly, the availability of detailed protocols is limited. Lung MMP-9 immunostaining is further complicated by technical issues such as edge effect, availability of positive and negative controls, antigen retrieval, staining specificity, and the need to achieve a delicate balance of primary and secondary antibody concentrations, and colorimetric reagents which will allow visualization of specific cell expression in highly delicate lung tissue, while also demonstrating adequate uptake in (extra-pulmonary) tissue controls. We describe a detailed method for immunostaining MMP-9 in mouse lung paraffin-embedded tissue utilizing human ovary as a control since MMP-9 is known to be over-expressed in human ovarian carcinomas. PMID- 21644820 TI - The risks of subconscious biases in drug-discovery decision making. PMID- 21644821 TI - Antivirulence agents against MRSA. PMID- 21644822 TI - Synthetic protocol published for promising anticancer compound. PMID- 21644823 TI - Bridging gaps in discovery and development: chemical and biological sciences for affordable health, wellness and sustainability. AB - To commemorate 2011 as the International Year of Chemistry, the Indian Society of Chemists and Biologists organized its 15th International Conference on 'Bridging Gaps in Discovery and Development: Chemical and Biological Sciences for Affordable Health, Wellness and Sustainability' at Hotel Grand Bhagwati, in association with Saurashtra University, Rajkot, India. Anamik Shah, President of the Indian Society of Chemists and Biologists, was organizing secretary of the conference. Nicole Moreau, President of the International Union of Pure and Applied Chemistry and Secretary General of the Comite National de la Chimie, National Centre for Scientific Research France, was chief guest of the function. The four-day scientific program included 52 plenary lectures, 24 invited lectures by eminent scientists in the field and 12 oral presentations. A total of 317 posters were presented by young scientists and PhD students in three different poster sessions. Approximately 750 delegates from India, the USA, UK, France, Switzerland, Germany, Austria, Belgium, Sweden, Japan and other countries attended the conference. The majority of the speakers gave presentations related to their current projects and areas of interest and many of the talks covered synthesis, structure-activity relationships, current trends in medicinal chemistry and drug research. PMID- 21644824 TI - The University of Kansas High-Throughput Screening laboratory. Part I: meeting drug-discovery needs in the heartland of America with entrepreneurial flair. AB - The University of Kansas High-Throughput Screening (KU HTS) core is a state-of the-art drug-discovery facility with an entrepreneurial open-service policy, which provides centralized resources supporting public- and private-sector research initiatives. The KU HTS core applies pharmaceutical industry project management principles in an academic setting by bringing together multidisciplinary teams to fill critical scientific and technology gaps, using an experienced team of industry-trained researchers and project managers. The KU HTS proactively engages in supporting grant applications for extramural funding, intellectual-property management and technology transfer. The KU HTS staff further provides educational opportunities for the KU faculty and students to learn cutting-edge technologies in drug-discovery platforms through seminars, workshops, internships and course teaching. This is the first instalment of a two part contribution from the KU HTS laboratory. PMID- 21644825 TI - Growing PAINS in academic drug discovery. AB - In a recent article it was argued that compounds published as drug leads by academic laboratories commonly contain functionality that identifies them as nonspecific 'pan-assay interference compounds' (PAINS). The article raises broad questions about why best practices for hit and lead qualification that are well known in industry are not more widely employed in academia, as well as about the role of journals in publishing manuscripts that report drug leads of little potential value. Barriers to adoption of best practices for some academic drug discovery researchers include knowledge gaps and infrastructure deficiencies, but they also arise from fundamental differences in how academic research is structured and how success is measured. Academic drug discovery should not seek to become identical to commercial pharmaceutical research, but we can do a better job of assessing and communicating the true potential of the drug leads we publish, thereby reducing the wastage of resources on nonviable compounds. PMID- 21644826 TI - Assessing directed evolution methods for the generation of biosynthetic enzymes with potential in drug biosynthesis. AB - To address the synthesis of increasingly structurally diverse small-molecule drugs, methods for the generation of efficient and selective biological catalysts are becoming increasingly important. 'Directed evolution' is an umbrella term referring to a variety of methods for improving or altering the function of enzymes using a nature-inspired twofold strategy of mutagenesis followed by selection. This article provides an objective assessment of the effectiveness of directed evolution campaigns in generating enzymes with improved catalytic parameters for new substrates from the last decade, excluding studies that aimed to select for only improved physical properties and those that lack kinetic characterization. An analysis of the trends of methodologies and their success rates from 81 qualifying examples in the literature reveals the average fold improvement for k (cat) (or V (max)), K (m) and k (cat)/K (m) to be 366-, 12- and 2548-fold, respectively, whereas the median fold improvements are 5.4, 3 and 15.6. Further analysis by enzyme class, library-generation methodology and screening methodology explores relationships between successful campaigns and the methodologies employed. PMID- 21644827 TI - Phosphoprotein phosphatase 2A: a novel druggable target for Alzheimer's disease. AB - Tau hyperphosphorylation is thought to play an important role in the etiology of Alzheimer's disease by facilitating the formation of neurofibrillary tangles. Reducing phosphorylation through kinase inhibition has therefore emerged as a target for drug development, but despite considerable efforts to develop therapeutic kinase inhibitors, success has been limited. An alternative approach is to develop pharmaceuticals to enhance the activity of the principal phospho tau phosphatase, phosphoprotein phosphatase 2A (PP2A). In this article we review evidence that this mechanism is pharmacologically achievable and has promise for delivering the next generation of Alzheimer's disease therapeutics. A number of different chemotypes have been reported to lead to enhanced PP2A activity through a range of proposed mechanisms. Some of these compounds appear to act directly as allosteric activators of PP2A, while others act indirectly by inhibiting the binding of PP2A inhibitors or by altering post-translational modifications that act in turn to regulate PP2A activity towards phospho-tau. These results indicate that PP2A may provide a useful target that can be safely, selectively and effectively modulated through pharmaceutical intervention to treat Alzheimer's disease. PMID- 21644829 TI - Welcome to the animal pharmaceuticals special focus. PMID- 21644828 TI - Towards novel radiosensitizing agents: the role of cytosolic PLA2alpha in combined modality cancer therapy. AB - The radioresistant nature of some tumors serves as an obstacle to curative therapy for several poor-prognosis malignancies. The radiosensitivity of a cancer is dependent not only on the intrinsic ability of tumor cells to recover from radiation-induced damage, but also the ability of stromal elements (e.g., vasculature) in the tumor microenvironment to survive and continue proliferating in the face of ionizing radiation. In this regard, it is important to understand the initial events activating radiation-induced signal transduction pathways. Among these events is the activation of cytosolic phospholipase A2 alpha and the subsequent production of the lipid second messengers. These events occur within minutes following exposure to ionizing radiation, and have been shown to enhance cell viability through a number of prosurvival signaling pathways. Furthermore, inhibition of cytosolic phospholipase A2 alpha has now been shown to reduce the viability of endothelial cells in culture after exposure to ionizing radiation, as well as slowing the growth of tumors in animal models of cancer. PMID- 21644830 TI - Intellectual property issues for veterinary pharmaceuticals in the USA. PMID- 21644831 TI - Animal-health pharmaceuticals: research responsibilities and efforts in target animal safety and laboratory animal welfare. AB - As researchers in animal health, we are charged with numerous responsibilities. Among the greatest of these are ensuring the safety and effectiveness of the products we develop and the appropriate use of animals in our research efforts. The following discussion focuses primarily on the demonstration of drug safety in the species for which the product is to be licensed or registered (target animal safety) in the USA, and on our role as stewards of animal welfare in laboratory research. PMID- 21644832 TI - Role of the cytochrome P450 enzyme system in veterinary pharmacokinetics: where are we now? Where are we going? AB - Drug metabolism is a core determinant of the dose-effectiveness-toxicity relationship of many compounds. It is also critical to the human food safety assessment of drug residues in the edible tissues of food-producing animals. This article describes the current state of knowledge regarding the role of the cytochrome P450 superfamily of enzymes in determining the metabolic profile of compounds administered to companion animals (e.g., dog and cat) and to food producing animal species (e.g., cattle, swine, chickens). In turn, this knowledge reflects the collection of insights derived from the recognized population variability observed in human drug metabolism, our general understanding of the kinetics of various drug-metabolism pathways, emerging tools that enable the role of pharmacogenetics to be studied and the characterization of drug metabolism in individual veterinary species. Ultimately, by increasing our insights with regard to factors that can influence drug metabolism, our knowledge of metabolic pathways, sources of within- and between-species variability in pharmacokinetics and the development of in silico models that can be used to predict pharmacokinetic profiles from these diverse sources of information. We will improve our ability to generate the population inferences needed to insure the target animal safety, product effectiveness and the human food safety of veterinary pharmaceuticals. PMID- 21644833 TI - Overview of the animal health drug development and registration process: an industry perspective. AB - Products for animal health commercialization follow a structured progression from initial concept through to regulatory approval. Typically, products are developed for use in either food animals or companion animals. These can be for the intention of disease intervention, productivity enhancement or improvement in a quality of life capacity. The animal health industry is a regulated industry, meaning that a government agency is responsible for oversight of products, both pre- and post-approval. There are three primary US government agencies that ensure quality, safety and effectiveness for the approval of new products and post-marketing compliance. PMID- 21644834 TI - Discovery and development of veterinary antiparasitic drugs: past, present and future. AB - Despite investment in programs to manage the development of resistance to existing agents, this continues to drive the need for discovery of novel antiparasitic agents for veterinary medicine. Historically, antiparasitic drug discovery was driven by empirical screening, but technological advances have lead to an increased focus on mechanism-based approaches to drug discovery and this is projected to increase as our capabilities advance to improve both the throughput of assays and the quality of data generated. Investment in the development of combination products with novel agents is increasing and, despite regulatory hurdles in some regions, efforts to globally harmonize regulations will aid in delivering safe, efficacious drugs to help in resistance management and integrated parasite control programs. PMID- 21644835 TI - Mitochondria as therapeutic targets for the treatment of malignant disease. AB - SIGNIFICANCE: Mitochondria exert vital functions during normal physiology and are also centrally involved in the regulation of various modes of cell death. Thus, engaging the mitochondrial apoptosis pathway presents an attractive possibility to activate lethal effectors in cancer cells. RECENT ADVANCES: Compounds that directly target mitochondria offer the advantage to initiate mitochondrial outer membrane permeabilization independently of upstream signal transduction elements that are frequently impaired in human cancers. As a consequence, mitochondrion targeted agents may bypass some forms of drug resistance. CRITICAL ISSUES: An ever-increasing number of compounds, including natural compounds and rationally designed drugs, has been shown to directly act on mitochondria. FUTURE DIRECTIONS: Forthcoming insights into the fine regulation of mitochondrial apoptosis will likely open future perspectives for cancer drug development. PMID- 21644836 TI - The Renin-angiotensin system and reactive oxygen species: implications in pancreatitis. AB - SIGNIFICANCE: The renin-angiotensin system (RAS) is a circulating hormonal system involved in the regulation of blood pressure and circulating fluid electrolytes. Recent findings have revealed that locally generated angiotensin (Ang) II plays a pivotal role in normal physiology as well as pathophysiology in various tissues and organs, including the pancreas. This review article summarizes current progress that has been made in elucidating the putative roles of Ang II in both acute and chronic pancreatitis. RECENT ADVANCES: A convergence of evidence suggests that the underlying mechanism may involve reactive oxygen species (ROS) generating systems, such as nicotinamide adenine dinucleotide phosphate oxidase, and subsequent elevation of proinflammatory and profibrogenic gene expression as well as protein activity. More importantly, Ang II-induced ROS interacts with other ROS-generating systems to positively feed-forward the ROS-induced signaling. CRITICAL ISSUES AND FUTURE DIRECTIONS: Advances in basic research indicate that RAS blockers may provide potential therapeutic role for the management of pancreatic inflammation and, more importantly, pancreatitis associated complications. Genetic alterations resulting from a malfunction in the epigenetic control of pancreatic RAS could be a causative factor in the development of pancreatitis. PMID- 21644837 TI - Effectiveness of piroxicam and ibuprofen premedication on orthodontic patients' pain experiences. AB - OBJECTIVE: To compare the effectiveness of preoperative administration of ibuprofen and piroxicam on orthodontic pain experienced after separator placement. MATERIALS AND METHODS: Ninety patients aged between 13 years 9 months and 18 years 2 months who were to undergo fixed appliance orthodontic treatment were enrolled in this double-blind, parallel-arm, prospective study. Patients were evenly and randomly distributed to any of three experimental groups, as follows: (1) administration of placebo, (2) administration of 400 mg ibuprofen, and (3) administration of 20 mg piroxicam; medications were administered 1 hour before separator placement. The pain perceived was recorded by the patients on a linear and graded Visual Analogue Scale at time intervals of 2 hours; 6 hours; nighttime on the day of appointment; 24 hours after the appointment; and 2 days, 3 days, and 7 days after separator placement during each of the four activities (viz, chewing, biting, fitting front teeth, and fitting back teeth). RESULTS: The results revealed that preoperative administration of 20 mg of piroxicam 1 hour prior to separator placement resulted in a significant decrease in pain levels at 2 hours, 6 hours, nighttime, and 24 hours and on the second and third days after separator placement, compared to patients on a placebo or ibuprofen. CONCLUSIONS: Premedication with 20 mg of piroxicam results in significantly decreased pain experienced, compared to premedication with 400 mg of ibuprofen or placebo. Usage of 20 mg of piroxicam 1 hour prior to separator placement is recommended. PMID- 21644838 TI - Scaffold/matrix attached region-based nonviral episomal vectors. AB - Because of their high efficiency, virus-based vectors are currently used in most gene therapy trials. Because such vectors bear some potential safety risks, nonviral expression systems could be an attractive alternative. Ideally, these vectors should be completely based on chromosomal elements and replicate as an autonomous unit in the recipient cell, thus avoiding the risk of insertional mutagenesis or immunological reactions of the recipient organism. Our limited knowledge of the epigenetic regulation of replication in mammalian cells does not yet allow the rational design of such constructs. But in the late 1990s it was shown that scaffold/matrix attached region (S/MAR)-based vectors can promote episomal replication and maintenance in mammalian cells. These vectors have found broad application in basic research but are now improved for their use in the safe and reproducible genetic modification of cells and organisms and in gene therapy. PMID- 21644839 TI - Melampsora larici-populina transcript profiling during germination and timecourse infection of poplar leaves reveals dynamic expression patterns associated with virulence and biotrophy. AB - Melampsora larici-populina is responsible for poplar leaf rust disease and causes severe epidemics in poplar plantations in Europe. The poplar rust genome has been recently sequenced and, in order to find the genetic determinants associated with its biotrophic lifestyle, we generated a whole-genome custom oligoarray and analyzed transcript profiles of M. larici-populina during the infection timecourse in poplar leaves. Different stages were investigated during the asexual development of the rust fungus, including resting and germinating urediniospores and seven in planta stages in the telial host. In total, 76% of the transcripts were detected during leaf infection as well as in urediniospores, whereas 20% were only detected in planta, including several transporters and many small secreted proteins (SSP). We focused our analysis on gene categories known to be related to plant colonization and biotrophic growth in rust pathogens, such as SSP, carbohydrate active enzymes (CAZymes), transporters, lipases, and proteases. Distinct sets of SSP transcripts were expressed all along the infection process, suggesting highly dynamic expression of candidate rust effectors. In contrast, transcripts encoding transporters and proteases were mostly expressed after 48 h postinoculation, when numerous haustoria are already formed in the leaf mesophyll until uredinia formation, supporting their role in nutrient acquisition during biotrophic growth. Finally, CAZymes and lipase transcripts were predominantly expressed at late stages of infection, highlighting their importance during sporulation. PMID- 21644841 TI - The enhanced effect of atenolol on hypertension in metabolic syndrome. AB - BACKGROUND: In metabolic syndrome, hypertension has been noted as one of the most important risk factors that contributes to cardiovascular disease. We have evaluated the effect of a selective adrenergic beta(1)-receptor blocker, atenolol, on blood pressure and pulse rate in metabolic syndrome. METHODS: Metabolic syndrome patients with hypertension (n=28) and a control group (n=20) were given atenolol (50 mg/day) for 4 weeks. Blood pressure, pulse rate, and blood biochemical parameters were monitored. RESULTS: Pulse rate in the metabolic syndrome group before the treatment was significantly higher than in the control group (control 74 +/- 2/min, metabolic syndrome 84 +/- 2/min, P<0.05). Decrease in pulse rate resulting from the treatment was greater in the metabolic syndrome group than in the control group (control 14 +/- 3/min, metabolic syndrome 21 +/- 2/min, P<0.05). Decrease in systolic (SBP) and diastolic (DBP) blood pressure was greater in the metabolic syndrome group than in the control group (SBP, control 8 +/- 3, metabolic syndrome 23 +/- 3 mmHg, P<0.05; DBP, control 6 +/- 3, metabolic syndrome 13 +/- 2 mmHg, P<0.05). There was a positive association between decrease in SBP and the waist measurements of patients, between the decrease in DBP and the waist measurements, and between the decrease in pulse rate and the waist measurements. We also found that there was a positive association between the decrease in SBP and initial SBP, between the decrease in DBP and initial DBP, and between the decrease in pulse rate and initial pulse rate. CONCLUSIONS: These data showed that atenolol has an enhanced effect on blood pressure and on pulse rate in metabolic syndrome. It may be useful in treating severe hypertension with elevated heart rate in metabolic syndrome. PMID- 21644842 TI - Delivery approaches for angiogenic growth factors in the treatment of ischemic conditions. AB - INTRODUCTION: Despite current medical treatments, cardiovascular disease resulting in local ischemia remains a significant clinical problem. Therapeutic angiogenesis, that is, the growth and remodeling of new blood vessels from pre existing blood vessels to the ischemic area, is a promising solution to this problem. AREAS COVERED: Therapeutic angiogenesis can be generated in vivo through the local release of various proangiogenic factors. This review describes the various formulation approaches that have been devised for this purpose, highlighting the advantages and disadvantages of each. EXPERT OPINION: Formulations that release single proangiogenic growth factors have not yet been demonstrated to achieve functional therapeutic angiogenesis. Formulations capable of multiple growth factor delivery are needed; however, the complexity of the physiologic process requires the examination of appropriate growth factor doses, as well as release sequence, to guide effectively new formulation design. Furthermore, new formulation approaches need to be tested in vivo in appropriate animal models over extended time periods to assess clearly the potential of the delivery approach. PMID- 21644843 TI - Varenicline for tobacco dependence: panacea or plight? AB - INTRODUCTION: This review examines the postmarketing experience with varenicline, including case reports, newer clinical trials and secondary analyses of large clinical datasets. AREAS COVERED: Varenicline has been shown to be an effective treatment in a broad range of tobacco users with medical, behavioral and diverse demographic characteristics. Recent studies finding excellent safety and efficacy in groups of smokers with diseases including chronic obstructive pulmonary disease are particularly encouraging and call for increased use of this medication for smoking cessation. Despite case reports of serious neuropsychiatric symptoms in patients taking varenicline, including changes in behavior and mood, causality has not been established. Recent analyses of large datasets from clinical trials have not demonstrated that varenicline is associated with more depression or suicidality than other treatments for smoking cessation. EXPERT OPINION: Now that additional clinical trials in specific populations and observational studies on treatment-seeking smokers outside of clinical trials have been published, we can be confident that varenicline remains the most efficacious monotherapy for smoking cessation and that its side-effect profile remains good. The risk-to-benefit ratio of receiving varenicline to quit smoking must include the increased chances of quitting smoking and avoiding the sizeable risks of smoked-caused disease and death that remain if tobacco addiction is not properly treated. PMID- 21644844 TI - Review of the use of mirtazapine in the treatment of depression. AB - INTRODUCTION: Depression is a heterogeneous illness affecting large numbers of patients with far reaching effects on both psychiatric and medical outcomes as well as quality of life. Knowledge about the different treatment modalities used to treat depression is important to all physicians. AREAS COVERED: This article expands on one such treatment option, mirtazapine. The paper examines this medication's medicinal chemistry, pharmacological properties as well as the efficacy of the medication compared to other commonly used antidepressants. EXPERT OPINION: Mirtazapine is a medication unlike other agents used for depression both in its mechanism of action as well as its side effects. Knowing these distinct characteristics will enable physicians to better utilize this agent for the benefit of their patients. PMID- 21644845 TI - A pilot study of the impact of genotype on nifedipine pharmacokinetics when used as a tocolytic. AB - OBJECTIVE: To characterize the pharmacokinetics of nifedipine when used for tocolysis in preterm labor and to determine the impact of genetics on these parameters. STUDY DESIGN: Pharmacokinetic study performed on women given tocolytic nifedipine. Over one dosing interval, drug concentrations, clinical data, and genotype for Cytochrome P450 (CYP)3A5 polymorphisms were obtained. Non compartmental pharmacokinetic analysis was used to estimate nifedipine exposure at steady state. RESULTS: The mean nifedipine area under the curve in 20 pregnant women was 86.1+/-61.1 ng/ml/h. The mean nifedipine exposure differed by expression of CYP3A5 (expressers [exp]: 139.5+/-97.3 ng/ml/h vs. nonexpressers[non]: 68.3 +/- 31.8 ng/ml/h, p = 0.02). Four women consumed CYP3A inhibitors and this affected the nifedipine concentrations (p < 0.001). CYP3A5 expressers had less improvement in contraction frequency after the loading dose (p = 0.04), at steady state (p = 0.006), and at 0-1 h after the study dose (p < 0.001). CONCLUSIONS: CYP3A5 genotype plays a role in nifedipine concentration when used as a tocolytic. PMID- 21644846 TI - Testicular apoptosis after dietary zinc deficiency: ultrastructural and TUNEL studies. AB - The present study was conducted in Wistar rats to determine whether prepubertal dietary zinc deficiency causes apoptotic changes in testes. Prepubertal male Wistar rats (40-50 gm) were divided into 3 groups: zinc control (ZC), pairfed (PF), and zinc deficient (ZD). Control and pairfed groups were given a 100 ppm zinc diet while the deficient groups received 1 ppm zinc diet for 2 and 4 weeks (w), respectively. Ultrastructural studies revealed several apoptotic features such as wavy basement membrane, displaced nuclei, chromatin condensation, plasma membrane blebbing, nuclear membrane dissolution, loss of inter-Sertoli cell junctional complexes, and intercellular bridges and deformed mitochondria. A variable spectrum of sperm defects had also been visualized e.g., acrosomal deformities such as decapitation and a ring of condensed chromatin around the nuclear periphery, deformed sperm heads with a condensed nucleus, tail-elements with superfluous cytoplasm, and damage to the mitochondrial sheath and aggregation of spermatozoa within the membrane. This was further supported by TUNEL studies. Apoptotic index, epididymal sperm concentration, motility, and fertility index also revealed a significant (P < 0.05) decrease in zinc deficient groups (2 and 4 w) when compared with their respective control and pairfed groups. All the above findings are indicative that changes observed in the testes after dietary zinc deficiency are due to the onset of apoptosis. Increased apoptotic degeneration in testes may cause irreversible changes in the germ cells associated with decreased epididymal sperm concentration, motility, and fertility index which contributes to the low efficiency of spermatogenesis thereby indicating a possible role of zinc in fertility. PMID- 21644847 TI - Evaluation of vitamin D levels among HIV-infected patients in New York City. AB - Few studies have examined the vitamin D status in HIV-infected patients. A cross sectional retrospective chart review of 2992 HIV-infected patients was conducted from 9/2008 to 5/2009. A total of 274 adult patients had 25-hydroxyvitamin D [25(OH)D] obtained by radioimmunoassay. None was receiving vitamin D (vitD) supplements. Vitamin D status was defined as the following: vitD deficiency (vitDd) as 25(OH)D <25 nmol/liter, vitD insufficiency (vitDi) as 25(OH)D 25-74 nmol/liter, and vitD optimal (vitDo) as 25(OH)D >=75 nmol/liter. We analyzed demographic/laboratory data. vitDd, vitDi, and vitDo were 21.2% (58 patients, 58/274), 68.6% (188 patients, 188/274), and 10.2% (28 patients, 28/274), respectively. There were significant racial differences. Blacks were 60.3% (35 patients, 35/58), 40.4% (76 patients, 76/188), and 28.6 % (8 patients, 8/28) in vitDd, vitDi, and vitDo, respectively, p=0.002. CD4 T cell count was not different in these three groups. However, HIV viral load was significantly different. Median log (10) HIV viral load was 2.31 with IQR 1.70-409, 1.70 with IQR 1.70-2.96, and 1.70 with IQR 1.70-2.78 in vitDd, vitDi, and vitDo, respectively, p=0.039. Multivariate logistic regression analysis showed that black race [odd ratio (OR) 4.108, 95% confidence interval (CI) 1.462-11.543, p=0.007] and HIV viral load>50 copies/ml (OR 2.396, 95% CI 1.120-5.127, p=0.024) were significantly associated with vitamin D deficiency. Vitamin D deficiency was highly prevalent in HIV-infected patients. Detectable HIV viremia and dark skin (black ethnicity) were significantly associated with vitamin D deficiency. Evaluation of vitamin D status in HIV-infected patients should be considered and further studies are needed to define the effects of vitamin D. PMID- 21644848 TI - Intramural hematoma of the esophagus as a complication of anticoagulant or antiplatelet therapy. PMID- 21644849 TI - Longitudinal associations between cycling to school and weight status. AB - OBJECTIVE: The objective of the present study was to assess the longitudinal association between cycling to school and weight status in two cities where cycling to school is common - Kristiansand (Norway) and Rotterdam (The Netherlands). METHODS: Data from two studies ENDORSE (Rotterdam) and YOUTH IN BALANCE (Kristiansand) were used. Both studies were longitudinal with two years between time (T) 1 and T2 measurements, and with mean age at T1 of 13.2 and 13.4 years, respectively. The sample was categorized into the following groups according to responses about main mode of commuting to school at the two time points: NO cycling, STARTED cycling, STOPPED cycling and CONTINUED cycling. Measured weight and height were obtained at both time points, and weight status (overweight vs. not overweight) was calculated using international classification criteria for BMI. The two datasets were analyzed separately and together. RESULTS: In multilevel logistic regression models of the combined sample, adjusting for weight status at Time 1, those who stopped cycling had greater odds of being overweight at T2 (OR = 3.19; 95% CI = 1.41?7.24) while those continued cycling had lower odds of being overweight (OR = 0.44; 95% CI = 0.21?0.88), separately compared to the other three groups together. The same trend was observed in both study samples. CONCLUSIONS: This study shows that there are longitudinal associations between cycling to school and weight status in two cities where cycling to school is common, implying that interventions aiming at reducing overweight/obesity among adolescents might consider the promotion of sustained cycling behaviour. PMID- 21644850 TI - The crucial role of early mitochondrial injury in L-lysine-induced acute pancreatitis. AB - AIMS: Large doses of intraperitoneally injected basic amino acids, L-arginine, or L-ornithine, induce acute pancreatitis in rodents, although the mechanisms mediating pancreatic toxicity remain unknown. Another basic amino acid, L-lysine, was also shown to cause pancreatic acinar cell injury. The aim of the study was to get insight into the mechanisms through which L-lysine damages the rat exocrine pancreas, in particular to characterize the kinetics of L-lysine-induced mitochondrial injury, as well as the pathologic responses (including alteration of antioxidant systems) characteristic of acute pancreatitis. RESULTS: We showed that intraperitoneal administration of 2 g/kg L-lysine induced severe acute necrotizing pancreatitis. L-lysine administration caused early pancreatic mitochondrial damage that preceded the activation of trypsinogen and the proinflammatory transcription factor nuclear factor-kappaB (NF-kappaB), which are commonly thought to play an important role in the development of acute pancreatitis. Our data demonstrate that L-lysine impairs adenosine triphosphate synthase activity of isolated pancreatic, but not liver, mitochondria. INNOVATION AND CONCLUSION: Taken together, early mitochondrial injury caused by large doses of L-lysine may lead to the development of acute pancreatitis independently of pancreatic trypsinogen and NF-kappaB activation. PMID- 21644851 TI - Supervision--growing and building a sustainable general practice supervisor system. AB - This article explores various models and ideas for future sustainable general practice vocational training supervision in Australia. The general practitioner supervisor in the clinical practice setting is currently central to training the future general practice workforce. Finding ways to recruit, retain and motivate both new and experienced GP teachers is discussed, as is the creation of career paths for such teachers. Some of the newer methods of practice-based teaching are considered for further development, including vertically integrated teaching, e learning, wave consulting and teaching on the run, teaching teams and remote teaching. Approaches to supporting and resourcing teaching and the required infrastructure are also considered. Further research into sustaining the practice based general practice supervision model will be required. PMID- 21644852 TI - General practice education and training: past experiences, current issues and future challenges. AB - Reflection on past achievements and future challenges 10 years after the establishment of the Australian General Practice Training program. PMID- 21644853 TI - The Australian General Practice Training program--reflections on the past decade. AB - How has general practice vocational training progressed towards the original goals established by the federal government and General Practice Education and Training 10 years ago? PMID- 21644854 TI - The evolution of general practice training in Australia. AB - Training for general practice in Australia has undergone a 60-year evolutionary process punctuated by revolutionary events. The discipline of general practice has also evolved significantly over this period. Today's Australian general practice training program strongly resembles its ancestors, with adaptations that better suit its regionalised environment. General practice training has been affected frequently by political and professional forces. Many of these forces were powered by the government's need for general practice training to deliver immediate workforce solutions, and the profession's struggle to respond. Pressure on general practitioners to train increasing numbers of clinical learners is challenging traditional apprenticeship models. The Australian general practice training program needs to continue to evolve if it is to remain successful within its volatile environment. PMID- 21644855 TI - Australian and overseas models of general practice training. AB - General practice training in Australia continues to evolve. It is now the responsibility of an independent organisation, is delivered by regional training providers, and comprises a structured training program. Overseas, general practice varies in its importance to health care systems, and training models differ considerably. In some cases training is mandatory, in others voluntary, but the aim is always similar--to improve the quality of care delivered to the large majority of populations that access health care through primary care. We review the current status of vocational general practice training in Australia, compare it with selected training programs in international contexts, and describe how the local model is well placed to address future challenges. Challenges include changes in population demographics, increasing comorbidity, increasing costs of technology-based health care, increasing globalisation of health, and workforce shortages. Although general practice training in Australia is strong, it can improve further by learning from other training programs to meet these challengers. PMID- 21644856 TI - General practice training in Aboriginal and Torres Strait Islander health. AB - This article reviews the history of general practice vocational training in Aboriginal and Torres Strait Islander health, identifies current initiatives and recommends future approaches based on recent evidence. General practice vocational training in Aboriginal and Torres Strait Islander health requires ongoing support and investment from governments and training and general practice organisations if the gains made to date are to be consolidated and health outcomes are to improve. In particular, investment in sustained and respectful partnerships with Aboriginal and Torres Strait Islander peoples and organisations will continue to provide the groundwork for effective training of general practitioners in this critical health area, and will also play an important role in capacity-building in Aboriginal and Torres Strait Islander communities. PMID- 21644857 TI - Regionalisation of general practice training--are we meeting the needs of rural Australia? AB - The concept of "social accountability" has underpinned the development of many medical education programs over the past decade. Success of the regionalisation of the general practice training program in Australia will ultimately be measured by the ability of the program to deliver a sufficient rural general practice workforce to meet the health needs of rural communities. Regionalisation of general practice training in Australia arose from the 1998 recommendations of the Ministerial Review of General Practice Training. The resultant competitive structure adopted by government was not the preferred option of the Review Committee, and may be a negative influence on rural workforce, as the competitive corporate structure of regional training providers has created barriers to meaningful vertical integration. Available data suggest that the regionalised training program is not yet providing a sustainable general practice workforce to rural Australia. The current increase in medical student and general practice training places provides an opportunity to address some of these issues. In particular, it is recommended that changes be made to registrar selection processes, the rural pipeline and vertical integration of training, and training for procedural rural practice. To achieve these goals, perhaps it is time for another comprehensive ministerial review of general practice training in Australia. PMID- 21644858 TI - Vertical integration of teaching in Australian general practice--a survey of regional training providers. AB - OBJECTIVE: To examine vertical integration of teaching and clinical training in general practice and describe practical examples being undertaken by Australian general practice regional training providers (RTPs). DESIGN, SETTING AND PARTICIPANTS: A qualitative study of all RTPs in Australia, mid 2010. RESULTS: All 17 RTPs in Australia responded. Eleven had developed some vertical integration initiatives. Several encouraged registrars to teach junior doctors and medical students, others encouraged general practitioner supervisors to run multilevel educational sessions, a few coordinated placements, linkages and support across their region. Three RTPs provided case studies of vertical integration. CONCLUSIONS: Many RTPs in Australia use vertical integration of teaching in their training programs. RTPs with close associations with universities and rural clinical schools seem to be leading these initiatives. PMID- 21644859 TI - Training Australian Defence Force Medical Officers to civilian general practice training standards--reflections on military medicine and its links to general practice education and training. AB - This article examines military medicine and its links to civilian general practice education and training, drawing attention to the variations and difficulties in, and successful approaches for, training Australian Defence Force (ADF) Medical Officers. Military medicine has been an area of change over the 10 years of the Australian General Practice Training (AGPT) program. Crisis situations like those in Timor Leste and Afghanistan have focused attention and recognition on the importance of primary health care in the work of the ADF. To train doctors in military medicine, there are several different models at different locations around Australia, as well as large variations in military course and experience recognition and approvals between AGPT regional training providers. At times, the lack of standardisation in training delays the progress of ADF registrars moving through the AGPT program and becoming independently deployable Medical Officers. PMID- 21644860 TI - Family medicine training--the international experience. AB - Family medicine is undergoing dramatic transformation around the world. Its organisation, delivery, and funding are changing in profound ways. While the specifics of primary care reform vary, a common emerging strategy involves establishment of primary health care teams that provide improved access, use electronic records, are networked with other teams, and are paid using blended payment schemes. More family doctors are needed in all countries. New approaches beyond the traditional apprenticeships or residency programs will be required to meet global demand. Training of family doctors must change to prepare tomorrow's family physician for a different practice reality. Curricula are more competency oriented, rather than time-focused. Today's trainees can anticipate a career that includes periodic reassessment of their knowledge base and competency. This article explores these trends and offers some strategies that have proved effective in various parts of the world for training increased numbers of qualified family doctors. PMID- 21644861 TI - Strategic approaches to the development of Australia's future primary care workforce. AB - Shortages in, and maldistribution of, the primary health care workforce will continue to limit access to health care. The current health reform proposals and policies recognise workforce development as a priority, but only partially address the barriers to improvement. In particular, there will need to be more systematic development of interdisciplinary education within primary health care services, and funding to support this. PMID- 21644862 TI - Getting governance right for a sustainable regionalised business model. AB - The 1998 Ministerial Review of General Practice Training identified several areas for improvement that led to major changes in the provision of general practice training, including the establishment of General Practice Education and Training (GPET) and the regionalisation of training. The regionalised training business model has been in place for nearly 10 years, and several key organisations have been involved in its evolution, including the Australian Government, speciality colleges, GPET and regionalised training providers. Both the college-focused and regionalised-focused models have had some successes. These include recognition and support of general practice as a vocational specialty, increased numbers of junior doctors undertaking placements in general practice, and increased numbers of registrars training in rural areas. This period has also seen changes in the governance and decision-making processes with creation of a new framework that is inclusive of all the key players in the new regionalised training system. The future holds challenges for the regionalised training business model as the general practice education and training landscape becomes more complex. The framework in the current model will provide a base to help meet these challenges and allow for further sustainable expansion. PMID- 21644863 TI - Future models of general practice training in Australia. AB - Current proposals for significant primary health care reform in Australia create a timely opportunity to reflect on the education and training requirements of future general practitioners. Australian general practice will become increasingly team-based, with growing emphasis on coordinated care, chronic disease management, and disease prevention and self-management, while maintaining its focus on delivering high-quality, patient-centred care. This will require cost-effective application of new technologies and information management systems within new models of delivering health care. Future models of general practice training must respond to these new ways of working to ensure general practice remains an attractive career choice and training programs graduate doctors who are equipped to meet the health needs of Australians. This article discusses potential development of new general practice vocational training models in Australia. This includes hospital rotations that are more directly integrated with general practice placements and have greater emphasis on the needs of the future general practice workforce; and an extension of the training program to 4 years with a final year tailored to future career plans including development of expertise in practice management, specific clinical disciplines or academic skills. PMID- 21644864 TI - Time to rethink end-of-life care. PMID- 21644865 TI - "Learning health care" for patients and populations. AB - A patient-focused learning health system, using integrated data sources, will facilitate optimal care of individuals and result in better care of our populations and society. PMID- 21644866 TI - Extensively resistant tuberculosis in the lands Down Under. AB - As tuberculosis resistance increases linearly, the cost and complexity of managing these cases increases exponentially. PMID- 21644867 TI - Saving money on the PBS: ranibizumab or bevacizumab for neovascular macular degeneration? AB - The cost differential between these two drugs is no longer defensible. PMID- 21644868 TI - Hospital and emergency department use in the last year of life: a baseline for future modifications to end-of-life care. AB - OBJECTIVES: To describe hospital and emergency department use in the last year of life by people for whom death from cancer or one of another nine conditions was an expected outcome. DESIGN, PARTICIPANTS AND SETTING: Retrospective cross sectional study based on death registrations and morbidity data for 1071 Western Australians who died between 1 August 2005 and 30 June 2006. Decedents had an informal primary carer, did not live in residential aged care and died of a condition amenable to palliative care. MAIN OUTCOME MEASURES: Total number of hospital admissions; emergency presentations (with and without hospital admission); days spent in hospital by age group at death, sex, metropolitan or rural place of residence and cancer versus non-cancer diagnosis; proportion in hospital on any day in the last 365 days of life; time points of change in the last 365 days of life at which there was an increasing proportion of hospital admissions for those with cancer and non-cancer conditions. RESULTS: All but 4% of the decedents spent time in hospital with a marked increase in hospitalisations in the last 108 days of life for people who died of cancer and the last 83 days of life for people who died of non-cancer conditions. Those with cancer spent less time in hospital than those with other diagnoses. Seventy per cent of the cohort had at least one emergency presentation. On the last day of life, 61.5% of people were in hospital and 4.0% had been seen in emergency departments. CONCLUSIONS: Western Australian hospitals currently provide extensive and progressively greater care at the end of life. Identifying patterns of emergency and inpatient use for various disease trajectories will assist in the planning of appropriate services for people where death is an expected outcome. PMID- 21644869 TI - The challenges of population ageing: accelerating demand for emergency ambulance services by older patients, 1995-2015. AB - OBJECTIVE: To measure the growth in emergency ambulance use across metropolitan Melbourne since 1995, to measure the impact of population growth and ageing on these services, and to forecast demand for these services in 2015. DESIGN AND SETTING: A population-based retrospective analysis of Ambulance Victoria's metropolitan emergency ambulance transportation data for the period from financial year 1994-95 to 2007-08, and modelling of demand in the financial year 2014-15. MAIN OUTCOME MEASURES: Numbers and rates of emergency ambulance transportations. RESULTS: The crude annual rate of emergency transportations across all age groups increased from 32 per 1000 people in 1994-95 to 58 per 1000 people in 2007-08. The rate of transportation for all ages increased by 75% (95% CI, 62%-89%) over the 14-year study period, representing an average annual growth rate of 4.8% (95% CI, 4.3%-5.3%) beyond that explained by demographic changes. Patients aged >= 85 years were eight times (incident rate ratio, 7.9 [95% CI, 7.6 8.3]) as likely to be transported than those aged 45-69 years over this period. Forecast models suggest that the number of transportations will increase by 46% 69% between 2007-08 and 2014-15, disproportionately driven by increasing usage by patients aged >= 85 years. CONCLUSIONS: These findings confirm a dramatic rise in emergency transportations over the study period, beyond that expected from demographic changes. Rates increased across all age groups, but more so in older patients. In the future, such acceleration is likely to have major effects on ambulance services and acute hospital capacity. This calls for further investigation of underlying causes and alternative models of care. PMID- 21644870 TI - Assessment and management of latent tuberculosis infection in a refugee population in the Northern Territory. AB - OBJECTIVES: To assess the prevalence of latent tuberculosis infection (LTBI) in recently arrived refugees in the Northern Territory and to obtain comprehensive data for rates of treatment acceptance and completion for this condition. DESIGN, SETTING AND PARTICIPANTS: Prospective data collection and follow-up of all 471 newly arrived refugees seen at the Centre for Disease Control, NT refugee health clinic from February 2006 to January 2009. MAIN OUTCOME MEASURES: Rates of LTBI determined by tuberculin skin testing; subsequent assessment and treatment compared with local protocols. RESULTS: 458 of 465 eligible refugees were adequately assessed for LTBI, of whom 146 (31.9%) were diagnosed with LTBI. Older age, male sex and World Health Organization Eastern Mediterranean region of birth were associated with increased prevalences of LTBI. Of the refugees diagnosed with LTBI, 10 failed to attend for follow-up and 15 were not offered treatment. Isoniazid therapy was accepted by 93 of 121 refugees (76.9%), and 41 of these (44.1%) completed treatment. The most common reasons for discontinuation of therapy were medication-related side effects (most often gastrointestinal) and loss to follow-up. Increasing age was associated with failure to complete treatment. CONCLUSION: Outcomes of assessment and treatment for LTBI in newly arrived refugees in the NT are comparable to those for other target groups screened in developed countries. Loss to follow-up caused significant attrition in numbers, but complete data were obtained for a large proportion of eligible refugees. Most refugees who are offered treatment for LTBI accept, but less than half complete treatment. PMID- 21644871 TI - Aseptic insertion of central venous lines to reduce bacteraemia. AB - OBJECTIVE: To reduce the rate of central line-associated bacteraemia (CLAB). DESIGN: A collaborative quality improvement project in intensive care units (ICUs) to promote aseptic insertion of central venous lines (CVLs). A checklist was used to record compliance with all aspects of aseptic CVL insertion, with maximal sterile barrier precautions for clinicians ("clinician bundle") and patients ("patient bundle"). CLAB was identified and reported using a standard surveillance definition. PARTICIPANTS AND SETTING: Patients and clinicians in 37 ICUs in New South Wales, July 2007-December 2008. MAIN OUTCOME MEASURES: Compliance with aseptic CVL insertion; rates of CLAB. RESULTS: 10 890 CVL checklists were reviewed for compliance with the clinician and patient bundles: compliance with aseptic CVL insertion improved significantly (P < 0.001). The CLAB rate dropped from 3.0 to 1.2 per 1000 line-days (P < 0.001). Regardless of CVL type, the relative risk (RR) of CLAB in patients with CVLs inserted by clinicians not compliant with the clinician bundle was 1.62 times greater (95% CI, 1.1-2.4; P = 0.018) than the RR with CVLs inserted by clinicians compliant with both bundles. Compliance with both the bundles was associated with a 50% reduction in risk of CLAB (RR, 0.5; 95% CI, 0.4-0.8; P = 0.004). CONCLUSIONS: Compliance with all aspects of aseptic CVL insertion significantly reduces the risk of CLAB. A difficulty we experienced was that most ICUs lacked the organisation and staff to support quality improvement and audit. PMID- 21644872 TI - Multidisciplinary team response to a mass burn casualty event: outcomes and implications. AB - OBJECTIVES: To describe the characteristics of patients with burn injury admitted to a major trauma hospital in Melbourne following the Black Saturday bushfires of 7 February 2009, and to provide a detailed analysis of the hospital's response to the crisis. DESIGN, SETTING AND PARTICIPANTS: A retrospective chart review of ambulance and hospital records of patients admitted to the Victorian Adult Burns Service (VABS) at The Alfred Hospital (The Alfred) following the bushfires. MAIN OUTCOME MEASURES: Patient characteristics and outcomes: age, sex, total and full thickness body surface area burnt, type and site of burn, hospital and intensive care unit length of stay (LOS) and receipt of standard burn care practices. Estimated glomerular filtration rate, theatre time and LOS data for the bushfire cohort compared with corresponding data for historical cohorts from VABS and from a similar institution in New Zealand. RESULTS: Nineteen patients were admitted to VABS over the first 48 hours after the bushfires. Of these, nine patients were subsequently admitted to The Alfred's intensive care unit. Most patients (74%) were men with a mean age of 52.7 years (SD, 12.4 years). Seventeen patients (89%) underwent at least one surgical procedure, which resulted in 4355 minutes of theatre time for the bushfire cohort in the first week. Hospital LOS was similar for the bushfire and New Zealand cohorts. Compared with the VABS historical cohort, there was a higher incidence of abnormal renal function among the bushfire cohort patients. CONCLUSIONS: Although relatively few patients with severe burns were admitted to VABS, significant increases in resource allocation were required to manage them in terms of additional theatre time, consumables and staffing. The experience of VABS may aid planning for future mass burns casualty events. PMID- 21644873 TI - Were the budgetary reforms to the Better Access to Mental Health Care initiative appropriate?--yes. AB - Mental health researchers Jane Pirkis and Meredith Harris say the government got it right. PMID- 21644874 TI - Were the budgetary reforms to the Better Access to Mental Health Care initiative appropriate?--no. AB - Mental health policy expert Sebastian Rosenberg and psychiatrist Ian Hickie want more change. PMID- 21644875 TI - Clinical deterioration in the condition of patients with acute medical illness in Australian hospitals: improving detection and response. AB - Medical Assessment Units (MAUs) provide an opportunity for multidisciplinary staff to manage recently admitted acutely unwell patients with complex medical illnesses. We propose concerted development of robust mechanisms for identifying and managing patients whose condition is unstable as they move through hospital departments. Track, trigger and response (TTR) systems (eg, medical emergency team calls and early warning scores) have been introduced to hospital practice, but evidence for their effectiveness is, so far, incomplete. The current variation in TTR systems within and between hospitals impairs intersite comparisons. A range of outcome measures, including risk of physiological deterioration, mortality and projected hospital length of stay, could be usefully investigated by future intersite collaborative research. More deliberate, systematic, evidence-based design of "response" in TTR systems may help in identifying patients who need early attention from skilled medical staff. We need more uniform TTR systems, more research on TTR systems and more multisite research; MAUs are ideally situated to address this important area. PMID- 21644876 TI - Prepaid coordinated care for patients with diabetes: practices and patients bear the risks. AB - The Australian Government is planning to pilot a model of prepaid funding for coordinated care of patients with diabetes in general practice. Patients will register with a practice that undertakes to coordinate their care, and practices will manage pre-allocated funds to provide services instead of billing Medicare. Systems to manage prepaid funds in Australian general practice have not yet been developed. In the model that has been proposed, practices with a small register of patients will be at risk of overspending, which may threaten practice viability and patient services. If the initiative is to have integrity, all patient services should be paid from the prepaid funds and patients should only attend the practice with which they have registered. Risks should be delineated and contingency plans made explicit before practices and patients commit to the initiative. PMID- 21644877 TI - Extensively drug-resistant tuberculosis: New Zealand's first case and the challenges of management in a low-prevalence country. AB - In 2010, an immigrant from Burma was the first person to be diagnosed in New Zealand with extensively drug-resistant tuberculosis (XDR-TB). The strain of Mycobacterium tuberculosis is the most resistant reported to date in Australasia. Key difficulties of managing this disease in a low-prevalence country were delays from drug-susceptibility testing and in acquiring appropriate medicines, and a lack of evidence-based guidelines. Solutions are needed for New Zealand and the wider region as more cases of XDR-TB are likely to be encountered in the future. PMID- 21644878 TI - A national survey of general practitioners' experiences of patient-initiated aggression in Australia. AB - OBJECTIVE: To determine the prevalence of patient-initiated aggression toward general practitioners in Australia. DESIGN, SETTING AND PARTICIPANTS: A cross sectional national survey, conducted during February-May 2010, of 3090 GPs in 19 Divisions of General Practice, purposively sampled to represent urban, rural and remote areas. MAIN OUTCOME MEASURE: Proportion of GPs experiencing patient initiated aggression. RESULTS: Eight-hundred and four GPs returned completed surveys (response rate, 26.3%). In the previous 12 months, 58% of GPs had experienced verbal abuse and 18% had experienced property damage or theft. Very few GPs had experienced physical abuse (6%), stalking (4%), sexual harassment (6%) or sexual assault (0.1%). After controlling for other demographic variables, GPs with fewer years of experience (P = 0.003), or who worked full-time or in larger practices (both P = 0.03) experienced significantly more verbal abuse than their counterparts, and GPs who worked full-time (P = 0.004) or in metropolitan areas (P = 0.01) experienced significantly more property damage or theft. Female GPs experienced significantly more sexual harassment than male GPs (P < 0.001). CONCLUSIONS: This is the first national evidence of the prevalence of patient aggression toward GPs in Australia, which could inform the development of policies and guidelines that aim to reduce the prevalence of patient aggression toward GPs. PMID- 21644879 TI - Anticonvulsant hypersensitivity syndrome: a rare and serious complication. PMID- 21644880 TI - Death and morbidity from supratherapeutic dosing of colchicine. PMID- 21644881 TI - Increasing prevalence of obesity in Australia and its possible effect on the occurrence of pulmonary thromboembolism. PMID- 21644882 TI - High-dose intravenous flucloxacillin may affect warfarin therapy. PMID- 21644883 TI - Lower-alcohol, lower-calorie wines: harm reduction or harm production? PMID- 21644884 TI - Life-threatening hypokalaemia associated with ibuprofen-induced renal tubular acidosis. PMID- 21644885 TI - Lessons from the 4-hour standard in England for Australia. PMID- 21644886 TI - Increased mortality associated with after-hours and weekend admission to the intensive care unit: a retrospective analysis. PMID- 21644887 TI - Questions and answers in Indigenous health. PMID- 21644888 TI - Our doctors making a difference. AB - Aboriginal and Torres Strait Islander doctors walking in both worlds for the benefit of all Australians. PMID- 21644889 TI - The challenges of remote area medical education. AB - To what extent should we cushion the realities of remote area living for young people who are seeking challenge and inspiration? PMID- 21644890 TI - Safeguard or mollycoddle? An exploratory study describing potentially harmful incidents during medical student placements in Aboriginal communities in Central Australia. AB - OBJECTIVE: To assess the number and characteristics of potentially harmful incidents occurring during placement of medical students in remote Aboriginal communities in the Northern Territory. DESIGN, PARTICIPANTS AND SETTING: A retrospective audit of medical students' files from Northern Territory General Practice Education placements in Central Australia for the period from January 2006 to December 2007. MAIN OUTCOME MEASURES: Number and type of potentially harmful incidents. RESULTS: A total of 163 placements were undertaken. Of these, 98 (60%) had adequate documentation to determine whether an incident had occurred. There were 28 cases (17%) where potentially harmful incidents were judged to have occurred. Most incidents fell under several descriptive categories, but clinical supervision, professional practice and administrative issues were most common. CONCLUSIONS: One in six students experienced a potentially harmful incident during remote area placement in 2006-2007. While acknowledging the exploratory nature of this investigation and the major educational benefits that clearly arise from these placements, our findings indicate problems with clinical supervision and administration. PMID- 21644891 TI - How can Australia do better for Indigenous health? AB - Respect, tolerance and trust in Aboriginal and Torres Strait Islander people are needed from government to improve the health and wellbeing of Indigenous Australians. PMID- 21644892 TI - Increasing alcohol restrictions and rates of serious injury in four remote Australian Indigenous communities. AB - OBJECTIVE: To document rates of serious injuries in relation to government alcohol restrictions in remote Australian Indigenous communities. DESIGN AND SETTING: An ecological study using Royal Flying Doctor Service injury retrieval data, before and after changes in legal access to alcohol in four remote Australian Indigenous communities, Queensland, 1 January 1996-31 July 2010. MAIN OUTCOME MEASURES: Changes in rates of aeromedical retrievals for serious injury, and proportion of retrievals for serious injury, before and after alcohol restrictions. RESULTS: After alcohol restrictions were introduced in 2002-2003, retrieval rates for serious injury dropped initially, and then increased in the 2 years before further restrictions in 2008 (average increase, 2.34 per 1000 per year). This trend reversed in the 2 years after the 2008 restrictions (average decrease, 7.97 per 1000 per year). There was a statistically significant decreasing time trend in serious-injury retrieval rates in each of the four communities for the period 2 years before the 2002-2003 restrictions, 2 years before the 2008 restrictions, and the final 2 years of observations (2009-2010) (P < 0.001 for all four communities combined). Overall, serious-injury retrieval rates dropped from 30 per 1000 in 2008 to 14 per 1000 in 2010, and the proportions of serious-injury retrievals decreased significantly for all four communities. CONCLUSION: The absolute and the proportional rates of serious injury retrievals fell significantly as government restrictions on legal access to alcohol increased; they are now at their lowest recorded level in 15 years. PMID- 21644893 TI - Alcohol restrictions in Indigenous communities: necessary but not sufficient. PMID- 21644894 TI - Alcohol restrictions in Indigenous communities: an effective strategy if Indigenous-led. PMID- 21644895 TI - Research, information and consent for the Australian Health Survey: a separate standard for Indigenous people? AB - In the next Australian Health Survey, Indigenous people under 18 years of age will be excluded from direct clinical measurements and laboratory tests. Indigenous people of all ages were to be excluded from the opportunity, offered to other Australians, to donate blood and urine samples to a national repository. This component has now been abandoned for the whole cohort. This sets perilous precedents of exclusion from opportunities available to all other Australians, and deprives the medical community of information that could inform strategies to improve health profiles and outcomes in this seriously disadvantaged group. PMID- 21644896 TI - Indigenous participation in the Australian Health Survey: a response. AB - The Australian Health Survey will deliver key health measures for all Australian children and adults, and collect information across Australia. The Australian Bureau of Statistics has consulted widely on the development of the Survey, and has shaped the survey according to strong and consistent advice in relation to Aboriginal and Torres Strait Islander peoples. It is hoped that this approach will maximise survey response rates and place us in good trust to embark on the next survey round. PMID- 21644897 TI - Social determinants and the health of Indigenous Australians. AB - Health is dependent on conditions that enable people to live lives they would choose to live. PMID- 21644898 TI - Glycaemia and albuminuria as predictors of coronary heart disease in Aboriginal and Torres Strait Islander adults: a north Queensland cohort. AB - OBJECTIVE: To evaluate the contribution of non-traditional risk factors to coronary heart disease (CHD) incidence in Indigenous adults. DESIGN, SETTING AND PARTICIPANTS: Cohort study of 1706 Aboriginal and Torres Strait Islander adults from 26 remote communities in far north Queensland who were initially free of CHD, with a mean of 7.5 years of follow-up. MAIN OUTCOME MEASURES: CHD-related deaths and hospitalisations obtained by record matching. RESULTS: CHD incidence was similar in men and women and in Aboriginals and Torres Strait Islanders; overall incidence was 12.1 (95% CI, 10.1-14.1) events per 1000 person-years. At baseline, prevalence of diabetes was 12.4% in Aboriginals and 22.3% in Torres Strait Islanders, prevalence of any albuminuria was similarly high (33.5%) in both groups, and participants with diabetes were 5.5 (95% CI, 4.2-7.3) times more likely to have albuminuria than those without diabetes. At follow-up, adjusted hazard ratios for CHD were 1.7 (95% CI, 1.01-2.8) for obesity based on waist circumference; 1.5 (95% CI, 1.01-2.3) for hypertension; 1.4 (95% CI, 0.9-2.2) for previous or current smoking; 1.9 (95% CI, 1.3-2.7) for elevated triglycerides; 1.3 (95% CI, 0.9-1.9) for low high-density lipoprotein cholesterol; 1.3 (95% CI, 0.8-2.2) for impaired fasting glucose; 2.4 (95% CI, 1.7-3.5) for diabetes; and 4.6 (95% CI, 2.9-7.1) for macroalbuminuria. Baseline albuminuria without diabetes increased risk by 50% (adjusted rate ratio, 1.5 [95% CI, 0.9-2.4]) but diabetes with macroalbuminuria amplified risk sixfold (adjusted rate ratio, 5.9 [95% CI, 3.4-10.1]). CONCLUSION: High prevalence of glycaemia and albuminuria in this population, especially when combined, account for much of the excess CHD risk beyond the traditional Framingham risk factors. They can be measured simply, lend themselves to cardioprotective interventions, and should be used routinely to estimate risk and monitor effectiveness of treatment. PMID- 21644899 TI - Sepsis in the tropical Top End of Australia's Northern Territory: disease burden and impact on Indigenous Australians. AB - OBJECTIVE: To describe the clinical and epidemiological features of sepsis and severe sepsis in the population of the tropical Top End of the Northern Territory of Australia and compare these with published estimates for temperate Australia, the United States and Europe. DESIGN, SETTING AND PARTICIPANTS: Prospective cohort study in the major hospital for tropical NT, a region where 27% of the population are Indigenous. We screened all adult (>= 15 years) acute hospital admissions over a 12-month period (6 May 2007-5 May 2008) for sepsis by standard criteria, and collected standardised clinical data. MAIN OUTCOME MEASURES: Population-based incidence of community-onset sepsis and severe sepsis requiring intensive care unit (ICU) admission; 28-day mortality rate and microbial epidemiology. RESULTS: There were 1191 hospital admissions for sepsis in 1090 patients, of which 604 (50.7%) were Indigenous people; the average age was 46.7 years. The age-adjusted annual population-based incidence of sepsis was 11.8 admissions per 1000 (mortality rate, 5.4%), but for Indigenous people it was 40.8 per 1000 (mortality rate, 5.7%). For severe sepsis requiring ICU admission, the incidence was 1.3 per 1000 per year (mortality rate, 21.5%), with an Indigenous rate of 4.7 per 1000 (mortality rate, 19.3%). CONCLUSIONS: The incidence of sepsis in the tropical NT is substantially higher than that for temperate Australia, the United States and Europe, and these differences are mainly accounted for by the high rates of sepsis in Indigenous people. The findings support strategies to improve housing and access to health services, and reduce comorbidities, alcohol and tobacco use in Indigenous Australians. The burden of sepsis in indigenous populations worldwide requires further study to guide appropriate resourcing of health care and preventive strategies. PMID- 21644900 TI - Epidemiology of syphilis in Australia: moving toward elimination of infectious syphilis from remote Aboriginal and Torres Strait Islander communities? AB - OBJECTIVE: To describe the epidemiology of infectious syphilis among Aboriginal and Torres Strait Islander (Indigenous) people in Australia. DESIGN AND SETTING: We assessed trends in national infectious syphilis notification rates from 2005 to 2009 using Poisson regression, with a focus on geographic and demographic differences by Indigenous status. We compared Indigenous and non-Indigenous rate ratios over the study period and summarised the annual changes (summary rate ratio). MAIN OUTCOME MEASURES: Crude notification rates and summary rate ratios by Indigenous status, jurisdiction, sex, age group and area of residence. RESULTS: From 2005 to 2009, in the Indigenous population, there was a substantial decline in the notification rate for infectious syphilis nationally; as well as in the following subgroups: females, 15-29 year olds, and people living in outer regional and remote areas in the Northern Territory and Queensland. In contrast, there was a significant (P < 0.001) upward trend in the notification rate in the non-Indigenous population nationally; as well as in males, in people aged 20 years and over, and in residents of metropolitan and regional areas, New South Wales, Queensland, South Australia, Victoria and Western Australia. The highest summary rate ratios were seen in remote/very remote areas (86.33; 95% CI, 57.45 129.74), in 15-19 year olds (64.65; 95% CI, 51.12-81.78), in females (24.59; 95% CI, 19.73-30.65), and in Western Australia (23.89; 95% CI, 19.82-28.82). CONCLUSION: These data demonstrate that Australia has two distinct patterns of infectious syphilis: a substantially declining occurrence in Indigenous remote communities and an increasing incidence in males residing in urban and regional areas. Given the decline in notification rates in Indigenous remote communities, now might be the right time to move toward eliminating infectious syphilis from Indigenous communities. PMID- 21644901 TI - Reducing the burden of cancer for Aboriginal and Torres Strait Islander Australians: time for a coordinated, collaborative, priority-driven, Indigenous led research program. AB - Australia's efforts to prevent, diagnose and treat cancer are not as successful for Aboriginal and Torres Strait Islander people as they are for other Australians. There is a need for a nationally coordinated, collaborative, priority-driven research effort to better understand what works, and we need to implement that knowledge. All aspects of the process must involve genuine Indigenous leadership and participation. PMID- 21644902 TI - The Lowitja Institute: building a national strategic research agenda to improve the health of Aboriginal and Torres Strait Islander peoples. AB - With community involvement, research can be a powerful tool for closing the gap in Indigenous health disparity. PMID- 21644903 TI - Indigenous child health checks: the view from the city. PMID- 21644904 TI - Use of eye care services by Indigenous Australian adults. PMID- 21644905 TI - Alfie the tooth fairy (an animation). PMID- 21644906 TI - Talking about hepatitis. PMID- 21644907 TI - Strong body, strong mind, strong culture. PMID- 21644908 TI - Mental illness or spiritual illness: what should we call it? PMID- 21644909 TI - Closing gaps, maintaining cadence and removing trampolines: a personal reflection on 20 years in health. PMID- 21644910 TI - Racism as a determinant of social and emotional wellbeing for Aboriginal Australian youth. AB - OBJECTIVE: To explore the associations between self-reported racism and health and wellbeing outcomes for young Aboriginal Australian people. DESIGN, SETTING AND PARTICIPANTS: A cross-sectional study of 345 Aboriginal Australians aged 16 20 years who, as participants in the prospective Aboriginal Birth Cohort Study, were recruited at birth between 1987 and 1990 and followed up between 2006 and 2008. MAIN OUTCOME MEASURES: Self-reported social and emotional wellbeing using a questionnaire validated as culturally appropriate for the study's participants; recorded body mass index and waist-to-hip ratio. RESULTS: Self-reported racism was reported by 32% of study participants. Racism was significantly associated with anxiety (odds ratio [OR], 2.18 [95% CI, 1.37-3.46]); depression (OR, 2.16 [95% CI, 1.33-3.53]); suicide risk (OR, 2.32 [95% CI, 1.25-4.00]); and poor overall mental health (OR, 3.35 [95% CI, 2.04-5.51]). No significant associations were found between self-reported racism and resilience or any anthropometric measures. CONCLUSIONS: Self-reported racism was associated with poor social and emotional wellbeing outcomes, including anxiety, depression, suicide risk and poor overall mental health. PMID- 21644911 TI - The transformative potential of young motherhood for disadvantaged Aboriginal and Torres Strait Islander women in Townsville, Australia. AB - OBJECTIVE: To explore attitudes to pregnancy and parenthood among a group of Indigenous young people in Townsville, Australia. DESIGN AND PARTICIPANTS: Mixed methods and a cross-sectional design involving Indigenous women from a Young Mums Group designing the research instruments and acting as peer interviewers. Data were collected in 2004 from young Indigenous people who had never been pregnant (171 students at three high schools and 15 people at a homeless youth shelter) using a computer-assisted self-administered survey; from 59 of this group who also participated in single sex focus group discussions; and from 10 pregnant and parenting young women in individual semi-structured interviews. MAIN OUTCOME MEASURE: Self-reported attitudes and behaviour about aspirations, pregnancy and parenthood. RESULTS: Only eight of 186 young Indigenous people who had never been pregnant reported wanting to have a child as a teenager. Large proportions of this group of 186 reported idealised views about pregnancy, particularly young men, with 50.5% reporting that being a parent would always be enjoyable, and 62.6% reporting that being a mother or a father would not change their lives. Idealised views were associated with earlier sexual initiation (P = 0.001). Issues identified in the narratives of young mothers related to difficult backgrounds, pregnancy "just happening" to them, and the transformative impact of having a child on their lives and aspirations. CONCLUSIONS: Accurate parenting information may be necessary to address unrealistic views about parenting among Indigenous young people. Young Indigenous parents often come from extremely disadvantaged backgrounds, and becoming a parent may be the impetus for positive change. PMID- 21644912 TI - Maternal smoking and smoking in the household during pregnancy and postpartum: findings from an Indigenous cohort in the Northern Territory. AB - OBJECTIVE: To describe the trends in maternal smoking and smoking in the household for a cohort of Indigenous women followed from late pregnancy to 7 months postpartum. DESIGN AND SETTING: Prospective cohort study embedded within a randomised controlled trial (RCT) performed in the Northern Territory involving participants recruited between 30 June 2006 and 4 May 2010. PARTICIPANTS: 215 Indigenous women aged 17-39 years who had been recruited into the RCT, 162 of whom had completed their last study visit at 7 months postpartum by 1 June 2010. MAIN OUTCOME MEASURES: Smoking status of women, and smoking within their households, in their third trimester, and at 1 month, 2 months and 7 months postpartum. RESULTS: There were complete data on women's smoking status for 121 participants. Among these, the self-reported smoking rate was 45% (95% CI, 36% 55%) during pregnancy, increasing to 63% (95% CI, 54%-71%) at 7 months postpartum. Of the 66 women who were non-smokers at the antenatal visit, 23 (35%; 95% CI, 23%-47%) were smoking by the time their baby reached 7 months of age. Thirty-one per cent (95% CI, 23%-39%) of households included people who smoked inside during the antepartum period, whereas 16% (95% CI, 10%-23%) included people who smoked inside at 7 months postpartum. CONCLUSIONS: While an apparent reduction in indoor exposure to tobacco smoke during the postpartum period is encouraging, this is offset by an increase in the proportion of antenatal non smokers who subsequently reported smoking after the birth of their child. More health care service delivery and research attention needs to be directed to smoking during pregnancy and to postpartum relapse in this population. PMID- 21644913 TI - The evidence for automated grading in diabetic retinopathy screening. AB - Systematic screening for diabetic retinopathy using retinal photography has been shown to reduce the incidence of blindness among people with diabetes. The implementation of diabetic retinopathy screening programmes faces several challenges. Consequently, methods for improving the efficiency of screening are being sought, one of which is the automation of image grading involving detection of images with either disease or of inadequate quality using computer software. This review aims to bring together the available evidence that is suitable for making a judgement about whether automated grading systems could be used effectively in diabetic retinopathy screening. To do this, it focuses on studies made by the few centres who have presented results tests of automated grading software on large sets of patients or screening episodes. It also considers economic model analyses and papers describing the effectiveness of manual grading in order that the effect of replacing stages of manual grading by automated grading can be judged. In conclusion, the review shows that there is sufficient evidence to suggest that automated grading, operating as a disease / no disease grader, is safe and could reduce the workload of manual grading in diabetic retinopathy screening. PMID- 21644914 TI - The role of natural killer T (NKT) cells in the pathogenesis of type 1 diabetes. AB - In type 1 diabetes, a failure in the regulation of either innate or acquired immunity may be the cause of autoimmune response. A cell population that may have a regulatory role of the immune response are the Natural Killer T (NKT) cells, which are a population expressing T lymphocyte antigen receptor (TCR), and are a common marker for NK cells. A distinctive characteristic in NKT cells is their capacity to produce large amounts of immune-modulating cytokines. A decrease in the number and/or functional incapability of NKT cells is associated with progression of type 1 diabetes and with other self-immune diseases. However, the relevance of such findings is not completely understood. Limitations of the current studies include the existing methods to measure NKT activation and the lack of assessment of the expression of genes affected by NKT action. Nevertheless, the study of NKT cells may be a new clinical approach to detect individuals at risk for having type 1 diabetes. Additional studies are needed to evaluate the clinical value of this new predictive tool. PMID- 21644915 TI - Diabetes and cardiovascular disease following kidney transplantation. AB - Kidney transplantation is being performed more frequently for individuals with end stage renal disease (ESRD) due to improved survival and quality of life compared to long-term dialysis. Though rates decrease after transplant, cardiovascular disease (CVD) remains the most common cause of death after kidney transplant. New-onset diabetes after transplant (NODAT), a common complication following kidney transplantation, and pre-transplant diabetes both significantly increase the risk for CVD. Several other risk factors for CVD in kidney transplant recipients have been identified; however, optimal therapy for controlling the risk factors of CVD after kidney transplantation, including NODAT and pre-transplant diabetes, is not well defined. In the following review we will discuss the role of traditional and non-traditional risk factors in CVD after kidney transplant and the mechanisms involved therein. We will also examine the current literature regarding treatment of these risk factors for the prevention of CVD. Finally, we will review the current recommendations for pre- and post transplant cardiovascular evaluation and management. PMID- 21644916 TI - The extra-hematopoietic role of erythropoietin in diabetes mellitus. AB - Diabetes mellitus is a complex metabolic disorder characterized by chronic hyperglycemia and vascular complications, leading to significant morbidity and mortality. All forms of diabetes are ultimately related to insufficient functional pancreatic beta-cell mass to maintain euglycemia. In this context, the promotion of beta-cell survival and function is a fundamental issue of direct relevance to diabetes prevention and treatment. Although first identified as a hematopoietic factor that promotes erythropoiesis and erythrocyte survival, an accumulating body of evidence suggests that erythropoietin (EPO) may also exert cytoprotective effects on non-erythroid tissues, including the brain, kidney, blood vessels, and pancreatic beta cells. Recent reports have demonstrated the biological effects of EPO on the pancreatic beta cells and its potential preventative and therapeutic role in diabetes. This review will focus on the emerging extra-hematopoietic roles of EPO and its potential protective role in diabetes. PMID- 21644917 TI - Adipose triglyceride lipase: a new target in the regulation of lipolysis by insulin. AB - In adipose tissue, the primary physiological function of insulin is the suppression of lipolysis, the hydrolysis of stored fat. Mechanistically, insulin suppresses lipolysis both in transcriptional and post-transcriptional levels. Insulin signaling acutely inhibits beta-adrenergic signaling by decreasing intracellular cyclic AMP levels and the rate of lipolysis. Insulin also suppresses lipolysis by down-regulating the expression of the rate-limiting lipolytic enzyme, adipose triglyceride lipase or ATGL. In insulin resistance and type 2 diabetes, insulin mediated attenuation of lipolysis is impaired leading to an increased rate of lipolysis and increased release of free fatty acids (FFA) in the circulation. This is one of the potential mechanisms behind the development of hyperlipidemia and subsequent metabolic abnormalities in type 2 diabetes. In this article, we focus on the recent findings that highlight distinct molecular mechanisms by which insulin action is mediated and possible implications of the deregulation of these pathways in the pathophysiological context. PMID- 21644918 TI - Anticancer properties of flavonoids: roles in various stages of carcinogenesis. AB - High dietary intake of fruits and vegetables is consistently associated with a reduced risk of common human cancers. The specific mechanisms of action of most phytochemicals in cancer prevention are not yet clear but appear to be varied. One class of compounds currently under investigation is flavonoids, a large group of molecules with similar structure. Although their bioavailability is discussed, numerous in vitro and animal model data suggest that flavonoids modulate important cellular and molecular mechanisms related to carcinogenesis, a multistep process involving the transformation, survival, proliferation, invasion, angiogenesis, and metastasis of the tumor cells. Epidemiological studies confirmed that, among many flavonoids, apigenin, epigallocatechin gallate, delphinidin and genistein appear to be beneficial compounds in various stages of carcinogenesis. Therefore, according to in vitro and in vivo studies, this review attempts to increase our understanding about the preventive and therapeutic effects of these compounds to facilitate extrapolation of results from animal studies to human situations. PMID- 21644919 TI - Antistaphylococcal activity of novel salicylanilide derivatives. AB - This study examined the antibacterial properties of nineteen benzoxazole, isoniazid, ethionamide and salicylanilide derivatives against Staphylococcus aureus (S. aureus). It was found that three salicylanilide-derived compounds demonstrated antistaphylococcal activity: 5-Chloro-2-hydroxy-N-(4 (trifluoromethyl)phenyl)benzamide (5-Cl-4'-CF3- SAL), 4-chloro-2-(3 chlorophenylcarbamyoyl)phenyl)-2-(benzyloxycarbonylamino)propanoate (AIM31) and 4 chloro-2- (4-(trifluoromethyl)phenylcarbamoyl)phenyl acetate (AIM33). Investigation of the chemical structures of these three compounds and comparison with a non-inhibitory salicylanilide compound (i.e. 5,3'-diCl-SAL) illustrated that different combinations of chemical groups at defined positions on the salicylanilide core structure had a marked influence on antistaphylococcal activity. The most effective compound was AIM33 which inhibited staphylococcal growth and displayed an initial MIC value of 3.12 MUg ml(-1) and subsequent investigation revealed that an MIC as low as of 0.5 MUg ml(-1) was achievable. In this case, the dual presence of a trifluoromethyl group and an acetylated phenolic hydroxyl to the salicylanilide core structure led to greatly enhanced activity. PMID- 21644920 TI - Quercetin-phospholipid complex: an amorphous pharmaceutical system in herbal drug delivery. AB - Development of amphiphilic drug-lipid complexes is a potential approach for improving therapeutic efficacy of the drugs by increasing solubility, release profile and oral bioavailability. Quercetin (3, 3', 4', 5, 7 pentahydroxyflavone), a polyphenolic flavonoid, shows several biological effects like anti-inflammatory, anti-cancer, antiproliferative, antimutagenic and apoptosis induction but its use is limited due to its low aqueous solubility. To overcome this limitation, a value added phospholipid complex of quercetin was developed to improve its aqueous solubility for better absorption through the gastrointestinal tract and this might result in improved bioavailability. The quercetin-phospholipid complex thus prepared was evaluated for various physico chemical parameters like infra red spectroscopy (FT-IR), differential scanning calorimetry (DSC), X-ray powder diffractometry (XRPD), scanning electron microscopy (SEM) and solubility study. The In vitro antioxidant activity was also studied. The phospholipid complex of quercetin was found to be fluffy and porous with rough surface in SEM. FTIR, DSC and XRPD data confirmed the formation of phospholipid complex. The water solubility of quercetin was improved by 12 folds (from 3.44 MUg/ ml to 36.81 MUg/ ml) in the prepared complex. There was no statistical difference between the quercetin complex and quercetin in the In vitro anti-oxidant activity, indicating that the process of complexation did not adversely affect the bioactivity of the active ingredient. PMID- 21644921 TI - Lectin-immobilized fluorescent nanospheres for targeting to colorectal cancer from a physicochemical perspective. AB - The goal of this research is to develop an imaging agent that enables real-time and accurate diagnosis of small-sized colorectal cancer. Since colorectal cancer initially develops in the mucous membrane of the large intestine, a nonabsorbable colonoscopic imaging agent capable of being administered intracolonically was designed. The imaging agent is peanut agglutinin (PNA)-immobilized polystyrene nanospheres with surface poly(N-vinylacetamide) (PNVA) chains encapsulating coumarin 6. PNA is a targeting moiety that binds to beta-D-galactosyl-(1-3)-N acetyl-D-galactosamine, which is the terminal sugar of the Thomsen-Friedenreich antigen that is specifically expressed on the mucosal side of colorectal cancer cells. PNVA is immobilized with the aim of reducing nonspecific interactions between the imaging agent and normal tissues, because the initial tumor-derived change is very small throughout the entire large intestine. Coumarin 6 is encapsulated into nanosphere cores to provide endoscopically-detectable fluorescence intensity. It is anticipated that the intracolonically-administered imaging agent recognizes tumor-derived changes in the large intestinal mucosa with high affinity and specificity. Real-time and accurate diagnosis of small sized early colorectal cancer can be achieved through an imaging agent providing clear fluorescence contrast between normal and cancer tissues observed with a florescence endoscope. This review describes the design concept of this nanoprobe from a physicochemical perspective. PMID- 21644922 TI - Applications of polymeric nanocapsules in field of drug delivery systems. AB - Drug-loaded polymeric nanocapsules have exhibited potential applications in the field of drug delivery systems in recent years. This article entails the biodegradable polymers generally used for preparing nanocapsules, which include both natural polymers and synthetic polymers. Furthermore, the article presents a general review of the different preparation methods: nanoprecipitation method, emulsion-diffusion method, double emulsification method, emulsion-coacervation method, layer-by-layer assembly method. In addition, the analysis methods of nanocapsule characteristics, such as mean size, morphology, surface characteristics, shell thickness, encapsulation efficiency, active substance release, dispersion stability, are mentioned. Also, the applications of nanocapsules as carriers for use in drug delivery systems are reviewed, which primarily involve targeting drug delivery, controlled/sustained release drug delivery systems, transdermal drug delivery systems and improving stability and bioavailability of drugs. Nanocapsules, prepared with different biodegradable polymers, have received more and more attention and have been regarded as one of the most promising drug delivery systems. PMID- 21644923 TI - An in situ gelling buccal spray containing platelet lysate for the treatment of oral mucositis. AB - Oral mucositis is an inflammatory disease of the mucosa lining oral cavity which leads to atrophy of the epithelium and to its ulceration to form chronic lesions. Many studies, both in vitro and in vivo, have disclosed the effectiveness of growth factors derived from platelets to enhance cell proliferation, differentiation, chemotaxis, angiogenesis and extracellular matrix synthesis involved in the healing of tissues. Despite potential usefulness of growth factors, animal-derived or genetically engineered ones are currently scarcely available for regenerative therapies. The aim of the present work was to study an in situ gelling formulation to be delivered by a spraying device to the oral cavity affected by mucositis. A vehicle based on Poloxamer 407 (F127) and sodium alginate (LVG) was developed. An extemporaneous loading of the vehicle with PL was achieved. The formulation was able to quickly thermogelify at 34-35 degrees C with a viscosity at 8 degrees C suitable for spraying; moreover it was characterized by good mucoadhesive properties. ELISA assay evidenced that at time zero the growth factor PDGF AB was compatible with the vehicle. The in vitro wound healing test showed that the formulation enhanced cell growth as PL alone even after 72 h of contact without cell apoptosis. The overall results indicate that PL loaded in the in situ gelling F127 and LVG vehicle can be profitably employed to treat buccal mucositis. PMID- 21644924 TI - Inorganic nanoparticles for enhanced photodynamic cancer therapy. AB - Photodynamic therapy (PDT) in cancer treatment uses photosensitizers to generate singlet oxygen followed by photoirradiation. The efficacy of PDT is greatly determined by the dosimetry of activation light and the photosensitizer (PS), modulating the photodynamic reaction at depth in diseased tissue. Development of nano-formulated photosensitizer has emerged as a promising field because of the biocompatibility and the accessibility for multi-functionalization of nanoparticles. In this review, we summarize the contemporary progress in use of inorganic nanoparticles for improvement of PDT in cancer therapeutics. PMID- 21644925 TI - Innovative technologies for drug delivery applications: from biopolymers and nanocapsules to inorganic materials. PMID- 21644926 TI - SNPS of drug metabolic enzymes and personalized medicine: part I. PMID- 21644927 TI - Discovery of cellular substrates for protein kinase A using a peptide array screening protocol. AB - Post-translational modification of proteins is a universal form of cellular regulation. Phosphorylation on serine, threonine, tyrosine or histidine residues by protein kinases is the most widespread and versatile form of covalent modification. Resultant changes in activity, localization or stability of phosphoproteins drives cellular events. MS and bioinformatic analyses estimate that ~30% of intracellular proteins are phosphorylated at any given time. Multiple approaches have been developed to systematically define targets of protein kinases; however, it is likely that we have yet to catalogue the full complement of the phosphoproteome. The amino acids that surround a phosphoacceptor site are substrate determinants for protein kinases. For example, basophilic enzymes such as PKA (protein kinase A), protein kinase C and calmodulin-dependent kinases recognize basic side chains preceding the target serine or threonine residues. In the present paper we describe a strategy using peptide arrays and motif-specific antibodies to identify and characterize previously unrecognized substrate sequences for protein kinase A. We found that the protein kinases PKD (protein kinase D) and MARK3 [MAP (microtubule-associated protein)-regulating kinase 3] can both be phosphorylated by PKA. Furthermore, we show that the adapter protein RIL [a product of PDLIM4 (PDZ and LIM domain protein 4)] is a PKA substrate that is phosphorylated on Ser(119) inside cells and that this mode of regulation may control its ability to affect cell growth. PMID- 21644928 TI - ATF4 deficiency protects mice from high-carbohydrate-diet-induced liver steatosis. AB - Chronic feeding of HCD (high-carbohydrate diet) is one of the major contributors to the prevailing of metabolic diseases. ATF4 (activating transcription factor 4) has been shown to play an important role in the regulation of glucose metabolism and obesity development; however, it is unclear how ATF4(-/-) mice respond to HCD. In the present study, we show that 8 weeks of HCD results in significant higher accumulation of TAGs (triacylglycerols) in livers and impairment in glucose tolerance in ATF4(+/+) mice, but not in ATF4(-/-) mice, compared with those on a normal diet. Meanwhile, energy expenditure is further enhanced by HCD in ATF4(-/-) mice. Moreover, we show that ATF4 deficiency suppresses HCD-induced SCD1 (stearoyl-CoA desaturase 1) expression, furthermore, oral supplementation of the main product of SCD1 oleate (18:1) increases TAG accumulation in livers of ATF4(-/-) mice. Taken together, these results suggest that ATF4 deficiency is protective for HCD-induced hepatic steatosis and impairment of glucose tolerance and insulin sensitivity. Furthermore, the resistance to hepatic steatosis is at least in part due to suppression of SCD1 expression under HCD. PMID- 21644929 TI - Eighteen novel MHC class I A alleles identified in Vietnamese-origin cynomolgus macaques. AB - We report herein the identification of 18 novel Mafa-A alleles in cynomolgus macaques of Vietnamese origin. PMID- 21644930 TI - Identification of a novel allele, MICA*063N, in a Chinese lung cancer patient. AB - MICA*063N has one nucleotide different from MICA*027 at position 184 (C->T) in codon 62 (CAG->TAG), resulting in a premature stop codon in exon 2. PMID- 21644931 TI - Association of major histocompatibility complex class I chain-related gene A microsatellite polymorphism and hepatocellular carcinoma in South China Han population. AB - The major histocompatibility complex (MHC) class I-related gene A (MICA) is a stress-induced antigen and plays a key role in anti-infectious and anti-tumoral immune responses. In this study, we investigated the MICA transmembrane polymorphism in hepatocellular carcinoma (HCC) patients in South China Han population. We found that the MICA-A5.1 polymorphism, which gives rise to a truncated protein, was increased in HCC patients compared with healthy controls (P(c) = 0.036). Furthermore, we found that HCC patients with MICA-A5.1 polymorphism had higher serum levels of soluble MICA (sMICA). These results suggest that MICA-A5.1 polymorphism is associated with HCC patients in South China Han population and the MICA-A5.1 polymorphism may contribute to the development of HCC by promoting the release of sMICA to evade tumor immunosurveillance. PMID- 21644932 TI - A new HLA-DQB1 sequence, DQB1*02:01:04, discovered during an external quality assessment exercise. AB - HLA-DQB1*02:01:04 differs from DQB1*02:01:01 by one nucleotide (G>A) at position 303 in exon 2 resulting in a silent substitution (codon 69 - GAG >GAA), conserved glutamate. PMID- 21644933 TI - HLA-B35 correlates with a favorable outcome following adjuvant administration of an HLA-matched allogeneic melanoma vaccine. AB - This work presents survival data of 42 melanoma patients at high risk for disease recurrence who received an allogeneic melanoma vaccine composed of three cell lines, each matching at least one allele of the recipient's human leukocyte antigen (HLA)-A and -B loci. The 5-year overall survival (OS) rate and disease free survival (DFS) compared favorably with the standard interferon-alpha regimen. Interestingly, patients bearing HLA-B35 had significantly better OS and DFS (OS of 100% and DFS of 90% for HLA-B35 vs 56% and 23%, for the non-B35 patients). In contrast, patients expressing HLA-B07 did not fare well with the vaccine. Although the data include a relatively small cohort of patients, it strongly hints toward a correlation between HLA types and potential benefit from anticancer immunotherapy. PMID- 21644934 TI - Maternal emotional distress in pregnancy and delivery of a small-for-gestational age infant. AB - OBJECTIVE: To assess the association between maternal emotional distress in pregnancy and delivery of a small-for-gestational age offspring. DESIGN: A cohort study in pregnancy. SETTING: Fifty hospitals with a maternity ward in Norway during 1998-2008. POPULATION: Seventy-one thousand eight hundred and ninety-eight women with a singleton pregnancy in the Norwegian Mother and Child Cohort Study were followed from early pregnancy to delivery. METHODS: Information on presence of emotional distress was obtained through self-administered questionnaires in pregnancy weeks 17 and 30 and on birthweight, gestational length at delivery and sex by linkage to the Medical Birth Registry of Norway. Emotional distress was measured by short forms of the Hopkins Symptom Checklist-25. MAIN OUTCOME MEASURE: Small-for-gestational age offspring (birthweight <2.5th percentile). RESULTS: Being emotionally distressed at gestational weeks 17 and 30 was not significantly associated with subsequent delivery of a small-for-gestational age infant (adjusted odds ratio 1.16; 95% confidence interval 0.87-1.54). This estimate was adjusted for smoking in pregnancy, parity, diabetes, pre-eclampsia, body mass index, education and maternal age. CONCLUSIONS: Emotional distress during pregnancy was not associated with subsequent delivery of a small-for gestational age infant. PMID- 21644935 TI - Cervical nitric oxide release in Chlamydia trachomatis and high-risk human papillomavirus infection. AB - OBJECTIVE: To compare cervical fluid nitric oxide release in women with and without Chlamydia trachomatis and high-risk human papillomavirus infection (hrHPV). DESIGN: An open clinical study. SETTING: University Hospital of Helsinki. POPULATION: Thirty-nine women with (n=21) and without C. trachomatis (n=18). METHODS: Chlamydia trachomatis and/or hrHPV were studied by using specific RNA- and DNA-based tests. Levels of cervical fluid nitric oxide metabolite (NOx) were assessed by the Griess reaction. MAIN OUTCOME MEASURES: The difference in cervical fluid NOx between women with and without C. trachomatis and hrHPV. RESULTS: Fourteen (67%) C. trachomatis-infected women and three (17%) noninfected women had concomitant hrHPV. The level of cervical fluid NOx in women with C. trachomatis (median 37.5 MUmol/l, 95% confidence interval 26.1-50.9) was higher (p=0.02) than that in C. trachomatis-noninfected women (median 19.7 MUmol/L, 95% confidence interval 5.6-30.0). The presence of hrHPV did not associate with any difference in NOx levels between C. trachomatis-infected or noninfected women. CONCLUSIONS: Chlamydia trachomatis was associated with increased release of nitric oxide metabolites in the uterine cervix. This stimulus was stronger than that of hrHPV, because no additional rise in NOx was seen in women with concomitant C. trachomatis and hrHPV infection. PMID- 21644936 TI - Expression of matrix metalloproteinases and their inhibitors at the feto-maternal interface in unruptured ectopic tubal pregnancy. AB - OBJECTIVE: To investigate expression of matrix metalloproteinases (MMPs) and the tissue inhibitors of MMPs (TIMPs) at the feto-maternal interface and non implantation sites in unruptured tubal pregnancies. DESIGN: Prospective study. Setting. University teaching hospital. POPULATION: Eighteen patients with unruptured tubal pregnancy undergoing salpingectomy. METHODS: Immunohistochemistry was used to detect MMP-2, -9 and -14, and TIMP-1, -2 and -3 at the feto-maternal interface and non-implantation regions of the Fallopian tube. Serum levels of human chorionic gonadotropin (beta-hCG) were determined, and trophoblastic invasion was histologiclly classified as stage I when limited to the tubal mucosa and stage II when extending to the muscular layer. MAIN OUTCOME MEASURES: The relation between serum beta-hCG concentration with the depth of trophoblastic invasion into the tubal wall and differential expression of MMPs and TIMPs. RESULTS: A significant difference in the beta-hCG concentrations was seen between stage I and II invasion. Immunoreactivity for MMP 2, -9 and -14, and TIMP-1, -2 and -3 was primarily localized to cytotrophoblasts. At the implantation sites, the intensity of MMPs increased along the invasive pathway towards the maternal interstitium. The expression of MMP-2, MMP-14 and TIMP-3 was localized to the epithelium and smooth muscle cells of the Fallopian tube, while expression of MMP-9, TIMP-1 and TIMP-2 was weak or absent. CONCLUSIONS: Human chorionic gonadotropin correlated positively with invasion stage of trophoblasts. At ectopic implantation sites, the expression of MMPs gradually increased with increasing invasion depth of trophoblasts. TIMP-1 and TIMP-2 were only weakly expressed. The imbalance between expression of MMPs and TIMPs at the ectopic implantation sites may lead to the extensive destructive degradation of the extracellular matrix. PMID- 21644937 TI - Pregnancy following robot-assisted laparoscopic myomectomy in women with deep intramural myomas. AB - OBJECTIVE: To describe fecundity after robot-assisted laparoscopic myomectomy for deep intramural myomas. DESIGN: Prospective observational study. SETTING: University hospital. POPULATION: Women undergoing robot-assisted laparoscopic myomectomy. METHODS: Expanding on a previous prospective feasibility study, 31 consecutive women in whom a robot-assisted laparoscopic myomectomy was performed between April 2006 and July 2010 were included. The women, of whom 14 had known infertility, were selected for having symptomatic, deep intramural myomas with a possible impact on fertility. Using a prospective protocol, relevant peri operative and follow-up data were retrieved. MAIN OUTCOME MEASURES: Fertility and pregnancy outcome. RESULTS: The 31 women included had a median age of 35 years (range 28-42 years) and median body mass index of 22.0 kg/m(2) (range 20.1-24.7 kg/m(2) ). Fifteen (68%) of the 22 women with an active wish of conceiving had become pregnant at a median time of 10 months after surgery. A total of 18 pregnancies occurred, resulting in three miscarriages, two terminated pregnancies, 10 successful term deliveries and three ongoing pregnancies. The subgroup of 14 women with a known but otherwise unexplained infertility had a similar pregnancy rate (69%) and of those, 55% conceived naturally. The women who conceived naturally were on average eight years younger than the women becoming pregnant after in vitro fertilization, and all miscarriages occurred in an in vitro fertilization pregnancy. CONCLUSIONS: In women with symptomatic, deep intramural myomas and either otherwise unexplained infertility or myomas with a possible effect on conception, the pregnancy rate following robot-assisted laparoscopic myomectomy was 68%. PMID- 21644939 TI - Smoking cessation for patients on clozapine. PMID- 21644938 TI - Cognitive regulation of emotion in bipolar I disorder and unaffected biological relatives. AB - OBJECTIVE: We examined the use of particular cognitive strategies for regulating negative emotion in relation to mood and temperament in BD-I, unaffected relatives of bipolar patients (UR), and healthy controls (HC). METHOD: Participants were 105 patients with BD-I, 124 UR, and 63 HC; all participants completed the Cognitive Emotion Regulation Questionnaire (CERQ), the Depression Anxiety Stress Scales (DASS), and the Hypomanic Personality Scale (HPS). RESULTS: The BD-I group reported more frequent use of rumination, catastrophizing and self blame, and less frequent use of putting into perspective, in response to negative life events, relative to the UR and HC groups. In BD-I, more frequent use of rumination was associated with increased DASS and HPS scores. By contrast, within the UR group, more frequent use of catastrophizing and self-blame were associated with increased DASS and HPS scores. In all participants, less frequent use of adaptive cognitive reframing strategies (e.g. putting into perspective) were associated with increased DASS scores. CONCLUSION: Both BD-I and UR groups reported more frequent use of maladaptive regulatory strategies previously associated with depression. Emotion regulation strategies of catastrophizing, self-blame, and cognitive reframing techniques may be associated with vulnerability for mood disorders, with the latter active within the general population regardless of biological vulnerability to disorder. PMID- 21644940 TI - Total smoking cessation and clozapine treatment - a diabetogenic combination? PMID- 21644941 TI - The effect of social adjustment and attachment style on suicidal behaviour. AB - OBJECTIVE: Prior studies examining the relationship between social adjustment and suicidal ideation or behaviour have not examined attachment. This study examines the effect of attachment on the association between current social adjustment and suicide attempt risk. METHOD: Attachment, social adjustment, and history of suicide attempt were assessed in patients participating in research on major depressive disorder (N = 524). Suicide attempters and non-attempters were compared with attachment style and social adjustment using hierarchical logistic regression models. The two factor scoring method of the Adult Attachment Scale (secure vs. avoidant) was utilized as each measures unique aspects of attachment. RESULTS: Anxious attachment (OR = 1.33; 95% CI = 1.016-1.728; P = 0.038) but not overall social adjustment (P = 0.14) was associated with a history of a past suicide attempt when both attachment and social adjustment were assessed in the same model. Among subtypes of social adjustment, work adjustment was associated with past history of suicide attempt (OR = 1.25; 95%CI = 1.019-1.540; P = 0.033). As impairment in work adjustment increased by 1 unit, the likelihood of reporting a suicide attempt increased by approximately 25%. There was no interaction between anxious attachment and work adjustment (P = 0.81). CONCLUSION: Anxious attachment and work adjustment warrant further study as potential treatment targets in depressed suicidal patients. PMID- 21644942 TI - Schizophrenia disorders, substance abuse and prior offending in a sequential series of 435 homicides. AB - OBJECTIVE: To examine the relationship between committing homicide, the presence of schizophrenia, substance misuse and past criminality. METHOD: The study employed a data linkage design, using contacts recorded on two statewide databases, one of which recorded public mental health services contacts and the second of which recorded contacts with the police. The estimated rates of schizophrenia disorders, substance abuse and criminal convictions found among a population of 435 homicide offenders were contrasted with estimated rates in two composite comparison samples. RESULTS: Of the 435 offenders, 38 (8.7%) had been diagnosed with a schizophrenia disorder, which was RR 13.11 (95% CI 9.14-18.80) times more likely than a comparison sample. Rates of known substance abuse between homicide offenders with and without schizophrenia and community-dwelling residents with schizophrenia did not differ significantly. However, these rates were higher than those found in the general community. A similar pattern emerged for comparisons regarding offending histories between these same groups. CONCLUSION: The association between homicidal violence and having a schizophrenia disorder cannot be explained away simply on the basis of either comorbid substance abuse or prior criminal offending. PMID- 21644943 TI - Exome sequencing of two patients in a family with atypical X-linked leukodystrophy. AB - We encountered a family with two boys similarly showing brain atrophy with reduced white matter, hypoplasia of the brain stem and corpus callosum, spastic paralysis, and severe growth and mental retardation without speaking a word. The phenotype of these patients was not compatible with any known type of syndromic leukodystrophy. Presuming an X-linked disorder, we performed next-generation sequencing (NGS) of the transcripts of the entire X chromosome. A single lane of exome NGS in each patient was sufficient. Six potential mutations were found in both affected boys. Two missense mutations, including c.92T>C (p.V31A) in L1CAM, were potentially pathogenic, but this remained inconclusive. The other four could be excluded. Because the patients did not show adducted thumbs or hydrocephalus, the L1CAM change in this family can be interpreted as different scenarios. Personal genome analysis using NGS is certainly powerful, but interpretation of the data can be a substantial challenge requiring a lot of tasks. PMID- 21644944 TI - Added value of CAC in risk stratification for cardiovascular events: a systematic review. AB - BACKGROUND: Identification of individuals at high risk for cardiovascular disease (CVD) is important to initiate adequate treatment and to prevent future events. Moreover, identification of low-risk individuals is important to refrain from unneeded therapy. Current risk prediction models do not accurately predict the risk of CVD in individuals, and new markers have been sought to improve the risk assessment in individuals. Coronary artery calcification (CAC) is a marker of atherosclerosis that might improve current risk assessment when added to traditional risk factors. MATERIALS AND METHODS: We performed a systematic review on PubMed search (1 February 2011) on studies reporting on the added value of CAC in risk prediction in asymptomatic individuals. RESULTS: Of 39 publications on CAC and CVD, nine studies were carried out in asymptomatic individuals. All studies showed an increase in area under the curve ranging from 0.05 to 0.20 when CAC was added to the risk model. Four studies reported on improvements of individuals in low-, intermediate-, and high-risk categories. Addition of CAC to the risk model resulted in a net reclassification improvement ranging from 14% to 30%, meaning that CAC measurement reclassified a substantial proportion of individuals into correct risk categories. This improvement was most pronounced in those at intermediate Framingham risk. CONCLUSIONS: The available studies consistently showed that CAC scoring improves risk stratification in CVD risk categories when added to traditional risk factors only, especially among individuals at intermediate risk for CVD. Cost-effectiveness analyses together with a randomized controlled trial are needed before widespread introduction of CAC in clinical care. PMID- 21644945 TI - Eight criticisms not to make about group selection. AB - Group selection, which was once widely rejected as a significant evolutionary force, is now accepted by all who seriously study the subject. There is still widespread confusion about group selection, however, not only among students and the general public, but among professional evolutionists who do not directly study the subject. We list eight criticisms that are frequently invoked against group selection, which can be permanently laid to rest based upon current knowledge. Experts will always find something to critique about group selection, as for any important subject, but these eight criticisms are not among them. Laying them to rest will enable authors to openly use the term group selection without being handicapped during the review process. PMID- 21644946 TI - The concurrent evolution of cooperation and the population structures that support it. AB - The evolution of cooperation often depends upon population structure, yet nearly all models of cooperation implicitly assume that this structure remains static. This is a simplifying assumption, because most organisms possess genetic traits that affect their population structure to some degree. These traits, such as a group size preference, affect the relatedness of interacting individuals and hence the opportunity for kin or group selection. We argue that models that do not explicitly consider their evolution cannot provide a satisfactory account of the origin of cooperation, because they cannot explain how the prerequisite population structures arise. Here, we consider the concurrent evolution of genetic traits that affect population structure, with those that affect social behavior. We show that not only does population structure drive social evolution, as in previous models, but that the opportunity for cooperation can in turn drive the creation of population structures that support it. This occurs through the generation of linkage disequilibrium between socio-behavioral and population structuring traits, such that direct kin selection on social behavior creates indirect selection pressure on population structure. We illustrate our argument with a model of the concurrent evolution of group size preference and social behavior. PMID- 21644947 TI - Visualizing fitness landscapes. AB - Fitness landscapes are a classical concept for thinking about the relationship between genotype and fitness. However, because the space of genotypes is typically high-dimensional, the structure of fitness landscapes can be difficult to understand and the heuristic approach of thinking about fitness landscapes as low-dimensional, continuous surfaces may be misleading. Here, I present a rigorous method for creating low-dimensional representations of fitness landscapes. The basic idea is to plot the genotypes in a manner that reflects the ease or difficulty of evolving from one genotype to another. Such a layout can be constructed using the eigenvectors of the transition matrix describing the evolution of a population on the fitness landscape when mutation is weak. In addition, the eigendecomposition of this transition matrix provides a new, high level view of evolution on a fitness landscape. I demonstrate these techniques by visualizing the fitness landscape for selection for the amino acid serine and by visualizing a neutral network derived from the RNA secondary structure genotype phenotype map. PMID- 21644948 TI - Complexity, pleiotropy, and the fitness effect of mutations. AB - One of the assumptions underlying many theoretical predictions in evolutionary biology concerns the distribution of the fitness effect of mutations. Approximations to this distribution have been derived using various theoretical approaches, of which Fisher's geometrical model is among the most popular ones. Two key concepts in this model are complexity and pleiotropy. Recent studies have proposed different methods for estimating how complexity varies across species, but their results have been contradictory. Here, we show that contradictory results are to be expected when the assumption of universal pleiotropy is violated. We develop a model in which the two key parameters are the total number of traits and the mean number of traits affected by a single mutation. We derive approximations for the distribution of the fitness effect of mutations when populations are either well-adapted or away from the optimum. We also consider drift load in a well-adapted population and show that it is independent of the distribution of the fitness effect of mutations. We show that mutation accumulation experiments can only measure the effect of the mean number of traits affected by mutations, whereas drift load only provides information about the total number of traits. We discuss the plausibility of the model. PMID- 21644949 TI - Fitness cost of pheromone production in signaling female moths. AB - A secondary sexual character may act as an honest signal of the quality of the individual if the trait bears a cost and if its expression is phenotypically condition dependent. The cost of increasing the trait should be tolerable for individuals in good condition but not for those in a poor condition. The trait thus provides an honest signal of quality that enables the receiver to choose higher quality mates. Evidence for sex pheromones, which play a major role in shaping sexual evolution, inflicting a signaling cost is scarce. Here, we demonstrate that the amount of the major component of the pheromone in glands of Lobesia botrana (Lepidoptera) females at signaling time was significantly greater in large than in small females, that male moths preferred larger females as mates when responding to volatile signals, and small virgin females, but not large ones, exposed to conspecific pheromone, produced, when mated, significantly fewer eggs than nonexposed females. The latter indicates a condition-dependent cost of signaling. These results are in accordance with the predictions of condition dependent honest signals. We therefore suggest that female signaling for males using sex pheromones bears a cost and thus calling may serve as honest advertisement for female quality. PMID- 21644951 TI - Male cockroaches prefer a high carbohydrate diet that makes them more attractive to females: implications for the study of condition dependence. AB - Sexual selection is a major force driving the evolution of elaborate male sexual traits. Handicap models of sexual selection predict that male sexual traits should covary positively with condition, making them reliable indicators of male quality. However, most studies have either manipulated condition through varying diet quantity and/or caloric content without knowledge of specific nutrient effects or have correlated proxies of condition with sexual trait expression. We used nutritional geometry to quantify protein and carbohydrate intake by male cockroaches, Nauphoeta cinerea, and related this to sex pheromone expression, attractiveness, and dominance status. We found that carbohydrate, but not protein, intake is related to male sex pheromone expression and attractiveness but not dominance status. Additionally, we related two condition proxies (weight gain and lipid reserves) to protein and carbohydrate acquisition. Weight gain increased with the intake of both nutrients, whereas lipid reserves only increased with carbohydrate intake. Importantly, lipid accumulation was not as responsive to carbohydrate intake as attractiveness and thus was a less-accurate condition proxy. Moreover, males preferentially consumed high carbohydrate diets with little regard for protein content suggesting that they actively increase their carbohydrate intake thereby maximizing their reproductive fitness by being attractive. PMID- 21644950 TI - Gene genealogies reveal differentiation at sex pheromone olfactory receptor loci in pheromone strains of the European corn borer, Ostrinia nubilalis. AB - Males of the E and Z strains of the European corn borer Ostrinia nubilalis (Lepidoptera: Crambidae) are attracted to different blends of the same pheromone components. The difference in male behavioral response is controlled by the sex linked locus Resp. The two types of males have identical neuroanatomy but their physiological specificity is reversed, suggesting that variation at the periphery results in behavioral change. Differences in the olfactory receptors (ORs) could explain the strain-specific antennal response and blend preference. Gene genealogies can provide insights into the processes involved in speciation and allow delineation of genome regions that contribute to reproductive barriers. We used intronic DNA sequences from five OR-encoding genes to investigate whether they exhibit fixed differences between strains and therefore might contribute to reproductive isolation. Although two genealogies revealed shared polymorphism, molecular polymorphism at three genes revealed nearly fixed differences between strains. These three OR genes map to the sex chromosome, but our data indicate that the distance between Resp and the ORs is >20 cM, making it unlikely that variation in pheromone-sensitive OR genes is directly responsible for the difference in behavioral response. However, differences in male antennal response may have their origin in the selection of strain-specific alleles. PMID- 21644952 TI - Growth rate variation among passerine species in tropical and temperate sites: an antagonistic interaction between parental food provisioning and nest predation risk. AB - Causes of interspecific variation in growth rates within and among geographic regions remain poorly understood. Passerine birds represent an intriguing case because differing theories yield the possibility of an antagonistic interaction between nest predation risk and food delivery rates on evolution of growth rates. We test this possibility among 64 Passerine species studied on three continents, including tropical and north and south temperate latitudes. Growth rates increased strongly with nestling predation rates within, but not between, sites. The importance of nest predation was further emphasized by revealing hidden allometric scaling effects. Nestling predation risk also was associated with reduced total feeding rates and per-nestling feeding rates within each site. Consequently, faster growth rates were associated with decreased per-nestling food delivery rates across species, both within and among regions. These relationships suggest that Passerines can evolve growth strategies in response to predation risk whereby food resources are not the primary limit on growth rate differences among species. In contrast, reaction norms of growth rate relative to brood size suggest that food may limit growth rates within species in temperate, but not tropical, regions. Results here provide new insight into evolution of growth strategies relative to predation risk and food within and among species. PMID- 21644953 TI - Divergent patterns of age-dependence in ornamental and reproductive traits in the collared flycatcher. AB - Sexual ornaments are predicted to honestly signal individual condition. We might therefore expect ornament expression to show a senescent decline, in parallel with late-life deterioration of other characters. Conversely, life-history theory predicts the reduced residual reproductive value of older individuals will favor increased investment in sexually attractive traits. Using a 25-year dataset of more than 5000 records of breeding collared flycatchers (Ficedula albicollis) of known age, we quantify cross-sectional patterns of age-dependence in ornamental plumage traits and report long-term declines in expression that mask highly significant positive age-dependency. We partition this population-level age dependency into its between- and within-individual components and show expression of ornamental white plumage patches exhibits within-individual increases with age in both sexes, consistent with life-history theory. For males, ornament expression also covaries with life span, such that, within a cohort, ornamentation indicates survival. Finally, we compared longitudinal age dependency of reproductive traits and ornamental traits in both sexes, to assess whether these two trait types exhibit similar age-dependency. These analyses revealed contrasting patterns: reproductive traits showed within-individual declines in late-life females consistent with senescence; ornamental traits showed the opposite pattern in both males and females. Hence, our results for both sexes suggest that age-dependent ornament expression is consistent with life history models of optimal signaling and, unlike reproductive traits, proof against senescence. PMID- 21644954 TI - Inversely related aposematic traits: reduced conspicuousness evolves with increased toxicity in a polymorphic poison-dart frog. AB - Prevailing theory contends that aposematic coloration evolves in tandem with toxicity so that the evolution of increased toxicity will accompany the evolution of greater conspicuousness. Although variation in aposematic coloration within single species should be selectively constrained, because individuals varying from a predator-recognized warning signal will incur greater risk of predation, several species of poison-dart frogs display remarkable phenotypic variation. This variation may have evolved to match different levels of toxicity, and these species provide excellent opportunities to examine the evolution of aposematic coloration. Here, I test whether increased conspicuousness in the granular poison dart frog evolved in tandem with increased toxicity. Contrary to classical predictions, toxicity assays, spectral reflectance measurements, and phylogenetic reconstruction reveal that the less conspicuous color morphs are actually significantly more toxic than the brightest, most conspicuous phenotypes and that the more toxic, less-conspicuous form evolved from a less toxic, more conspicuous ancestor. Through gas chromatography--mass spectrometry analysis of toxin profiles, I traced the increase in toxicity in the less-conspicuous populations to an acquisition of specific alkaloids, some of which are proven convulsants. These results challenge the tenet that increased conspicuousness always evolves with increased toxicity and support the idea that once aposematism has been established in a species, phenotypic variation may evolve from brightness and toxicity becoming decoupled. PMID- 21644955 TI - Reproductive character displacement and signal ontogeny in a sympatric assemblage of electric fish. AB - The reproductive signals of two or more taxa may diverge in areas of sympatry, due to selection against costly reproductive interference. This divergence, termed reproductive character displacement (RCD), is expected in species-rich assemblages, where interspecific signal partitioning among closely related species is common. However, RCD is usually documented from simple two-taxon cases, via geographical tests for greater divergence of reproductive traits in sympatry than in allopatry. We propose a novel approach to recognizing and understanding RCD in multi-species communities--one that traces the displacement of signals within multivariate signal space during the ontogeny of individual animals. We argue that a case for RCD can be made if the amount of signal displacement between a pair of species after maturation is negatively correlated to distance in signal space before maturation. Our application of this approach, using a dataset of communication signals from a sympatric Amazonian assemblage of the electric fish genus Gymnotus, provides strong evidence for RCD among multiple species. We argue that RCD arose from the costs of heterospecific mismating, but interacted with sexual selection--favoring the evolution of conspicuous male signals that not only serve for mate-choice, but which simultaneously facilitate species recognition. PMID- 21644956 TI - Back-and-forth hermaphroditism: phylogenetic context of reproductive system evolution in subdioecious Daphne laureola. AB - Recent phylogenetic analyses of sexual reproductive systems supported the evolutionary pathway from hermaphroditism to dioecy via gynodioecy in different groups of angiosperms. In this study, we explore the evolution of sexual reproductive systems in Daphne laureola L. (Thymelaeaceae), a species with variation in reproductive system among population. Sequences from the ITS region of the nuclear ribosomal cistron and two plastid markers (psbA-trnH and ndhF) were analyzed and used to map the population reproductive system along the molecular phylogeny. Our results support D. laureola as a monophyletic lineage with three different clades within the Iberian Peninsula. The hermaphroditic populations belong to two different clades, whereas gynodioecy is ubiquitous but characteristic of the third clade, which grouped together all the North-Western Iberian populations sampled, including the apparently oldest haplotype sampled. Gynodioecy appears as the most likely basal condition of the 13 analyzed populations, but different evolutionary transitions in reproductive sexual system were traced within each D. laureola clade. Both ecological conditions and (meta)population dynamics may help explain plant reproductive system evolution at the microevolutionary scale. Phylogenetic studies in which the historical relationships between populations differing in reproductive system can be ascertained will help to clarify the process. PMID- 21644957 TI - The context-dependent effect of multiple paternity on effective population size. AB - Effective population size (N(e)) is important because it describes how evolutionary forces will affect a population. The effect of multiple sires per female on N(e) has been the subject of some debate, at the crux of which is the effects of monandry and multiple-paternity (MP) on male variance in reproductive success. In both mating systems, females mate with several males over their lifetimes, but sire offspring with one male at a time in the former and have several sires per clutch in the latter. First, I theoretically show that whether the annual male variance in reproductive success in an MP population is greater or less than that of a monandrous population depends on the distributions of within-clutch paternity. Then, I simulated different distributions of within clutch paternity under a range of parameters that characterize natural populations to show that an MP population can have an N(e) smaller or larger than that of a monandrous population with otherwise equal dynamics. The N(e(MP)):N(e(Monandry)) ratio increased with mating frequency and female variance in reproductive success, was equalized by long generation times, and was affected by the distribution of within-clutch paternities. The results of this model provide a unifying framework for the debate. PMID- 21644958 TI - Metapopulation structure of a seed-predator weevil and its host plant in arms race coevolution. AB - Although the importance of gene flow in the geographic structuring of host parasite interactions has been well discussed, little is known about how dispersal drives the spatial dynamics of other types of coevolutionary interactions in nature. We evaluated the roles of gene flow in the geographically structured processes of a predator-prey arms race involving a seed-predatory weevil with a long mouthpart and its host camellia plant with a thick fruit coat. Molecular genetic analyses showed that both weevil and camellia populations were structured at a spatial scale of several kilometers. Importantly, the spatial pattern of the migration of weevils, but not that of camellias, imposed significant effects on the geographic configuration of the levels of coevolutionary escalation. This result suggests that even if migration is limited in one species (camellia), local coevolution with the other species that migrates between neighboring localities (weevil) can reduce the interpopulation difference in the local adaptive optima of the former species. Thus, gene flow of a species potentially homogenizes the local biological environments provided by the species and thereby promotes the evolutionary convergence of its coevolving counterparts. Consequently, by focusing on coevolutionary interactions in natural communities, "indirect" effects of gene flow on the adaptive divergence of organisms could be identified. PMID- 21644959 TI - Migration-induced phenotypic divergence: the migration-selection balance of correlated traits. AB - Genetically correlated traits are known to respond to indirect selection pressures caused by directional selection on other traits. It is however unclear how local adaptation in populations diverging along some phenotypic traits but not others is affected by the joint action of gene flow and genetic correlations among traits. This simulation study shows that although gene flow is a potent constraining mechanism of population adaptive divergence, it may induce phenotypic divergence in traits under homogeneous selection among habitats if they are genetically correlated with traits under divergent selection. This correlated phenotypic divergence is a nonmonotonous function of migration and increases with mutational correlation among traits. It also increases with the number of divergently selected traits provided their genetic autonomy relative to the uniformly selected trait is reduced by specific patterns of genetic covariances: populations with lower effective trait dimensionality are more likely to generate very large correlated divergence. The correlated divergence is likely to be picked up by Q(ST)-F(ST) analysis of population genetic differentiation and be erroneously ascribed to adaptive divergence under divergent selection. This study emphasizes the necessity to understand the interaction between selection and the genetic basis of adaptation in a multivariate rather than univariate context. PMID- 21644960 TI - Evolutionary responses of dispersal distance to landscape structure and habitat loss. AB - It is generally well understood that some ecological factors select for increased and others for decreased dispersal. However, it has remained difficult to assess how the evolutionary dynamics are influenced by the spatio-temporal structure of the environment. We address this question with an individual-based model that enables habitat structure to be controlled through variables such as patch size, patch turnover rate, and patch quality. Increasing patch size at the expense of patch density can select for more or less dispersal, depending on the initial configuration. In landscapes consisting of high-quality and long-lived habitat patches, patch degradation selects for increased dispersal, yet patch loss may select for reduced dispersal. These trends do not depend on the component of life history that is affected by habitat quality or the component of life-history through which density-dependence operates. Our results are based on a mathematical method that enables derivation of both the evolutionary stable strategy and the stationary genotype distribution that evolves in a polymorphic population. The two approaches generally lead to similar predictions. However, the evolutionary stable strategy assumes that the ecological and evolutionary time scales can be separated, and we find that violation of this assumption can critically alter the evolutionary outcome. PMID- 21644961 TI - Speciation and dispersal along continental coastlines and island arcs in the Indo West Pacific turbinid gastropod genus Lunella. AB - Species trees were produced for the Indo-West Pacific (IWP) gastropod genus Lunella using MrBayes, BEAST, and *BEAST with sequence data from four genes. Three fossil records were used to calibrate a molecular clock. Eight cryptic species were recognized using statistical methods for species delimitation in combination with morphological differences. However, our results suggest caution in interpreting ESUs defined solely by the general mixed Yule Coalescent model in genera like Lunella, with lower dispersal abilities. Four almost entirely allopatric species groups were recovered that differ in ecology and distribution. Three groups occur predominantly along continental coastlines and one occurs on island arrays. Sympatric species occur only in the torquata and coronata groups along coastlines, whereas species in the cinerea group, distributed in two dimensional island arrays, occur in complete allopatry. Dispersal along island arcs has been important in the maintenance of species distributions and gene flow among populations in the cinerea group. The emergence of new islands and their eventual subsidence over geological time has had important consequences for the isolation of populations and the eventual rise of new species in Lunella. PMID- 21644962 TI - Evolutionary novelties and losses in geometric morphometrics: a practical approach through hominin molar morphology. AB - Geometric morphometric techniques may offer a promising methodological approach to analyze evolutionary novelties in a quantitative framework. Nevertheless, and despite continuous improvements to this methodology, the inclusion of novel features in these studies presents some difficulties. In the present study, different methods to explicitly include novel traits in geometric morphometric analyses are compared, including homology-free approaches, landmark-based approaches, and combinations of both techniques. The two-dimensional occlusal morphology of the lower second molar in multiple hominin species was chosen to evaluate these methods, as an example of an anatomical structure including one novelty: a distal fifth cusp is present in earlier hominins, and notably absent in many later Homo species. Results reveal that different approaches provide different results, highlighting that the design of the conformations of landmarks has a high impact on the inferred conclusions. Among diverse methods, a combined approach including landmarks, sliding semilandmarks, and only one landmark related to the studied novelty (an indicator of its absence or presence and of its size, when present), was able to directly discern structures with and without the novel feature, circumventing some of the methodological difficulties associated with these traits. This study demonstrates the ability of geometric morphometric techniques to investigate evolutionary novelties and explores the implications of different methods, providing a reference context for future studies. PMID- 21644963 TI - Immune defense in leaf-cutting ants: a cross-fostering approach. AB - To ameliorate the impact of disease, social insects combine individual innate immune defenses with collective social defenses. This implies that there are different levels of selection acting on investment in immunity, each with their own trade-offs. We present the results of a cross-fostering experiment designed to address the influences of genotype and social rearing environment upon individual and social immune defenses. We used a multiply mating leaf-cutting ant, enabling us to test for patriline effects within a colony, as well as cross colony matriline effects. The worker's father influenced both individual innate immunity (constitutive antibacterial activity) and the size of the metapleural gland, which secretes antimicrobial compounds and functions in individual and social defense, indicating multiple mating could have important consequences for both defense types. However, the primarily social defense, a Pseudonocardia bacteria that helps to control pathogens in the ants' fungus garden, showed a significant colony of origin by rearing environment interaction, whereby ants that acquired the bacteria of a foster colony obtained a less abundant cover of bacteria: one explanation for this pattern would be co-adaptation between host colonies and their vertically transmitted mutualist. These results illustrate the complexity of the selection pressures that affect the expression of multilevel immune defenses. PMID- 21644964 TI - Global analysis of genes involved in freshwater adaptation in threespine sticklebacks (Gasterosteus aculeatus). AB - Examples of parallel evolution of phenotypic traits have been repeatedly demonstrated in threespine sticklebacks (Gasterosteus aculeatus) across their global distribution. Using these as a model, we performed a targeted genome scan- focusing on physiologically important genes potentially related to freshwater adaptation--to identify genetic signatures of parallel physiological evolution on a global scale. To this end, 50 microsatellite loci, including 26 loci within or close to (<6 kb) physiologically important genes, were screened in paired marine and freshwater populations from six locations across the Northern Hemisphere. Signatures of directional selection were detected in 24 loci, including 17 physiologically important genes, in at least one location. Although no loci showed consistent signatures of selection in all divergent population pairs, several outliers were common in multiple locations. In particular, seven physiologically important genes, as well as reference ectodysplasin gene (EDA), showed signatures of selection in three or more locations. Hence, although these results give some evidence for consistent parallel molecular evolution in response to freshwater colonization, they suggest that different evolutionary pathways may underlie physiological adaptation to freshwater habitats within the global distribution of the threespine stickleback. PMID- 21644965 TI - Chromosome number and sex determination coevolve in turtles. AB - Although much progress has been achieved in understanding the genetic basis of adaptation, the drivers of genome evolution remain obscure. For instance, extensive variation among reptilian genomes continues largely unexplained, yet reptiles hold critical clues about vertebrate evolution. Turtles posses diverse chromosome numbers (2N = 28-66) derived from extensive genomic rearrangements, plus varied sex-determining mechanisms (genotypic and temperature-dependent). Here, we show that rates of evolution in turtle chromosome number are ~20-fold higher along phylogenetic branches where transitions between sex-determining mechanisms also occur, revealing a strong coevolution of these traits and making drift a less likely driver. Directional tests indicate that both traits evolved effectively in synchrony. These events occurred near global extremes in temperature shifts over the last 200 million years, although the role of climate change remains unknown at this point. Two alternative testable explanations for these patterns are proposed. First, selection for sex determination turnover may co-opt mechanisms (e.g., chromatin remodeling) favoring genomic rearrangements. Alternatively, chromosomal rearrangements underlying diploid number evolution may alter gene regulation enabling transitions in sex-determining mechanisms. Our data indicate that the evolution of sex determination is intimately linked to profound genomic changes underlying diploid number evolution, the ecological context of which remains intriguing. PMID- 21644966 TI - Testing Cheverud's conjecture for behavioral correlations and behavioral syndromes. AB - Recent research regarding correlations among behaviors--under the labels of behavioral syndromes and animal personalities--has typically assumed that phenotypic correlations between behaviors are representative of underling genetic correlations. However, for behaviors, the concordance between phenotypic and genetic correlations has not been rigorously examined. I tested this assumption using published estimates and found phenotypic and genetic correlations to be strongly related but found that the average absolute difference between the two was quite high and similar to that observed in other traits. Using absolute differences as the sole criterion, phenotypic correlations do not reliably estimate the magnitude of genetic correlations for behaviors, which is problematic for behavioral syndrome researchers. However, phenotypic correlations explained 75% of the variation in genetic correlations and their sign was typically the same as that of genetic correlations. This suggests that phenotypic correlations between behaviors reliably estimate the direction of underling genetic relationships and provide considerable information regarding the magnitude of genetic correlations. Thus, if researchers are careful about the questions they ask, phenotypic correlations between behaviors can be informative regarding underling genetic correlations and their evolutionary implications. PMID- 21644967 TI - Deviation from the line of least resistance does not exclude genetic constraints: a comment on Berner et al. (2010). AB - Berner et al. (2010) found that freshwater adaptation of three-spined sticklebacks had not followed the direction of maximal evolvability. Based on this, they suggested that ancestral variance structure has not appreciably biased adaptive diversification. We reanalyze their data to show that evolution has happened in directions of much larger than average evolvability, and we conclude that their data are consistent with an influence of ancestral variational constraints. PMID- 21644968 TI - Cost of multiple matings in female adders (Vipera berus). PMID- 21644969 TI - Health expenditure dynamics and years of U.S. residence: analyzing spending disparities among Latinos by citizenship/nativity status. AB - OBJECTIVE: We investigate health expenditure disparities between Latinos and non Latino whites by years of United States residence and citizenship/nativity status. DATA SOURCES: We link the Medical Expenditure Panel Survey and the National Health Interview Survey from 2000 to 2007. The sample consists of 31,514 Latinos and 76,021 white adults (18-64 years). STUDY DESIGN: The likelihood of any health spending, total health expenditure, and the out-of-pocket (OOP) share of health expenditure are our main dependent variables. We use two-part multivariate models to adjust for confounding factors. A stratified analysis by insurance status checks for the results' robustness. The decomposition technique is implemented to estimate the share of disparities that can be explained by observed and unobserved variables. PRINCIPAL FINDINGS: Latinos are much less likely to have any health spending (68 percent), total health expenditure (57 percent), and more likely to pay OOP (6 percent) compared with the white population. Overall, disparities narrow or disappear for naturalized Latinos the longer they stay in the country. Among noncitizen Latinos, disparities remain constant or decline slightly, but they remain large over time. CONCLUSIONS: Low health spending by foreign-born Latinos contributes to health expenditure disparities between Latinos and whites. Our findings provide preliminary evidence on health-spending convergence over time between foreign-born Latinos and that of whites. PMID- 21644970 TI - Understanding variations in Medicare Consumer Assessment of Health Care Providers and Systems scores: California as an example. AB - OBJECTIVE: To understand reasons why California has lower Consumer Assessment of Healthcare Providers and Systems (CAHPS) scores than the rest of the country, including differing patterns of CAHPS scores between Medicare Advantage (MA) and fee-for-service, effects of additional demographic characteristics of beneficiaries, and variation across MA plans within California. STUDY DESIGN/DATA COLLECTION: Using 2008 CAHPS survey data for fee-for-service Medicare beneficiaries and MA members, we compared mean case mix adjusted Medicare CAHPS scores for California and the remainder of the nation. PRINCIPAL FINDINGS: California fee-for-service Medicare had lower scores than non-California fee-for service on 11 of 14 CAHPS measures; California MA had lower scores only for physician services measures and higher scores for other measures. Adding race/ethnicity and urbanity to risk adjustment improved California standing for all measures in both MA and fee-for-service. Within the MA plans, one large plan accounted for the positive performance in California MA; other California plans performed below national averages. CONCLUSIONS: This study shows that the mix of fee-for-service and MA enrollees, demographic characteristics of populations, and plan-specific factors can all play a role in observed regional variations. Anticipating value-based payments, further study of successful MA plans could generate lessons for enhancing patient experience for the Medicare population. PMID- 21644971 TI - Axon-glial interactions in the central nervous system. PMID- 21644973 TI - Landscape nutrition: seeing the forest instead of the trees. AB - Recent theories suggest that herbivores forage across many scales and that foraging decisions are driven by the distribution of nitrogen. However, experimental tests of these predictions across large landscapes are rare and difficult. Pretorius et al. (2011) present an elegant experimental design to test how patch size, local nutrient density and total nutrient load are detected by foraging African elephants (Loxodonta africana) in the Colophospermum mopane shrub veld in South Africa. This experiment should serve as a model for investigations of how herbivores detect and respond to high nutrient patches of different size; it also raises questions for further research, such as the fate of high nutrient patches as elephants disperse nutrients from them in urine, faecal material and carcasses deposited elsewhere in the landscape. PMID- 21644972 TI - Visualization of 3D osteon morphology by synchrotron radiation micro-CT. AB - Cortical bone histology has been the subject of scientific inquiry since the advent of the earliest microscopes. Histology - literally the study of tissue - is a field nearly synonymous with 2D thin sections. That said, progressive developments in high-resolution X-ray imaging are enabling 3D visualization to reach ever smaller structures. Micro-computed tomography (micro-CT), employing conventional X-ray sources, has become the gold standard for 3D analysis of trabecular bone and is capable of detecting the structure of vascular (osteonal) porosity in cortical bone. To date, however, direct 3D visualization of secondary osteons has eluded micro-CT based upon absorption-derived contrast. Synchrotron radiation micro-CT, through greater image quality, resolution and alternative contrast mechanisms (e.g. phase contrast), holds great potential for non destructive 3D visualization of secondary osteons. Our objective was to demonstrate this potential and to discuss areas of bone research that can be advanced through the application of this approach. We imaged human mid-femoral cortical bone specimens derived from a 20-year-old male (Melbourne Femur Collection) at the Advanced Photon Source synchrotron (Chicago, IL, USA) using the 2BM beam line. A 60-mm distance between the target and the detector was employed to enhance visualization of internal structures through propagation phase contrast. Scan times were 1 h and images were acquired with 1.4-MUm nominal isotropic resolution. Computer-aided manual segmentation and volumetric 3D rendering were employed to visualize secondary osteons and porous structures, respectively. Osteonal borders were evident via two contrast mechanisms. First, relatively new (hypomineralized) osteons were evident due to differences in X-ray attenuation relative to the surrounding bone. Second, osteon boundaries (cement lines) were delineated by phase contrast. Phase contrast also enabled the detection of soft tissue remnants within the vascular pores. The ability to discern osteon boundaries in conjunction with vascular and cellular porosity revealed a number of secondary osteon morphologies and provided a unique 3D perspective of the superimposition of secondary osteons on existing structures. Improvements in resolution and optimization of the propagation phase contrast promise to provide further improvements in structural detail in the future. PMID- 21644974 TI - Accommodating environmental variation in population models: metaphysiological biomass loss accounting. AB - 1. There is a pressing need for population models that can reliably predict responses to changing environmental conditions and diagnose the causes of variation in abundance in space as well as through time. In this 'how to' article, it is outlined how standard population models can be modified to accommodate environmental variation in a heuristically conducive way. This approach is based on metaphysiological modelling concepts linking populations within food web contexts and underlying behaviour governing resource selection. Using population biomass as the currency, population changes can be considered at fine temporal scales taking into account seasonal variation. Density feedbacks are generated through the seasonal depression of resources even in the absence of interference competition. 2. Examples described include (i) metaphysiological modifications of Lotka-Volterra equations for coupled consumer-resource dynamics, accommodating seasonal variation in resource quality as well as availability, resource-dependent mortality and additive predation, (ii) spatial variation in habitat suitability evident from the population abundance attained, taking into account resource heterogeneity and consumer choice using empirical data, (iii) accommodating population structure through the variable sensitivity of life history stages to resource deficiencies, affecting susceptibility to oscillatory dynamics and (iv) expansion of density-dependent equations to accommodate various biomass losses reducing population growth rate below its potential, including reductions in reproductive outputs. Supporting computational code and parameter values are provided. 3. The essential features of metaphysiological population models include (i) the biomass currency enabling within-year dynamics to be represented appropriately, (ii) distinguishing various processes reducing population growth below its potential, (iii) structural consistency in the representation of interacting populations and (iv) capacity to accommodate environmental variation in space as well as through time. Biomass dynamics provide a common currency linking behavioural, population and food web ecology. 4. Metaphysiological biomass loss accounting provides a conceptual framework more conducive for projecting and interpreting the population consequences of climatic shifts and human transformations of habitats than standard modelling approaches. PMID- 21644975 TI - Using social networks to deduce whether residents or dispersers spread parasites in a lizard population. AB - 1. Heterogeneity of host behaviour can play an important role in the spread of parasites and pathogens around wildlife populations. Social networks have previously been suggested to represent transmission pathways within a population, but where the dynamics of host-parasite interactions are difficult to observe, networks may also be used to provide insights into transmission processes. 2. Pygmy bluetongue lizards, Tiliqua adelaidensis, occupy individual territories, live exclusively in burrows constructed by spiders in Australian native grasslands and are hosts to a tick, Bothriocroton hydrosauri, and a nematode, Pharyngodon wandillahensis. 3. On five monthly occasions, the locations of all individual lizards in three study plots were used to construct weighted, undirected networks based on proximity of adjacent burrows. 4. The networks were used to explore alternative hypotheses about the spread of each parasite through the population: that stable population members that remained in the same burrow over the study period played a major role in influencing the pattern of infection or that dispersing individuals played a more significant role. 5. For ticks, host individuals that were infected were more connected in the network than uninfected hosts and this relationship remained significant for connections to residents in the population, but not for connections to dispersers. 6. For nematodes, infected and uninfected hosts did not differ in their overall strength of connection in the network, but infected hosts were more connected to dispersers than were uninfected hosts, suggesting that lizards moving across the population are the major agents for the transmission of nematodes. 7. This study shows how network analyses can provide new insights into alternative pathways of parasite spread in wildlife populations, where it is difficult to make direct observations of transmission-related behaviours. PMID- 21644976 TI - The nested assembly of individual-resource networks. AB - 1. Much of the current understanding of ecological systems is based on theory that does not explicitly take into account individual variation within natural populations. However, individuals may show substantial variation in resource use. This variation in turn may be translated into topological properties of networks that depict interactions among individuals and the food resources they consume (individual-resource networks). 2. Different models derived from optimal diet theory (ODT) predict highly distinct patterns of trophic interactions at the individual level that should translate into distinct network topologies. As a consequence, individual-resource networks can be useful tools in revealing the incidence of different patterns of resource use by individuals and suggesting their mechanistic basis. 3. In the present study, using data from several dietary studies, we assembled individual-resource networks of 10 vertebrate species, previously reported to show interindividual diet variation, and used a network based approach to investigate their structure. 4. We found significant nestedness, but no modularity, in all empirical networks, indicating that (i) these populations are composed of both opportunistic and selective individuals and (ii) the diets of the latter are ordered as predictable subsets of the diets of the more opportunistic individuals. 5. Nested patterns are a common feature of species networks, and our results extend its generality to trophic interactions at the individual level. This pattern is consistent with a recently proposed ODT model, in which individuals show similar rank preferences but differ in their acceptance rate for alternative resources. Our findings therefore suggest a common mechanism underlying interindividual variation in resource use in disparate taxa. PMID- 21644977 TI - Spatio-temporal dynamics of bumblebee nest parasites (Bombus subgenus Psythirus ssp.) and their hosts (Bombus spp.). AB - 1. A 39-year bumblebee data base was used to study the codistribution of six cuckoo bumblebees in the subgenus Psythirus of Bombus (hereafter called Psythirus) and their free-living bumblebee hosts in the British Isles. 2. A model of nest parasitism predicted host threshold densities and stable deterministic dynamics, with fluctuations only emerging as a result of environmental or demographic stochasticity. 3. Standardized transects indicated that variation in total number of records could be largely attributed to variation in observer effort; analyses were therefore carried out using relative abundance. 4. Spatially, parasite-free zones were evident in areas of low host abundance, but the host threshold for parasite presence differed among species and locations. 5. Temporally, the relative numbers of the parasite and host species remained relatively constant, except that the nest parasite P. campestris declined significantly since 1990. 6. There were consistent negative effects of the parasitic species on the numbers of hosts in the following year, and this pattern was seen over large geographic areas. 7. The spatio-temporal patterns confirmed a high degree of host specificity, except that P. campestris may be parasitizing not only B. pascuorum but also other species in the subgenus Thoracobombus. PMID- 21644978 TI - Introduced pathogens follow the invasion front of a spreading alien host. AB - 1. When an invasive species first colonizes an area, there is an interval before any host-specific natural enemies arrive at the new location. Population densities of newly invading species are low, and the spatial and temporal interactions between spreading invasive species and specific natural enemies that follow are poorly understood. 2. We measured infection rates of two introduced host-specific pathogens, the entomophthoralean fungus Entomophaga maimaiga and the baculovirus Lymantria dispar nucleopolyhedrovirus (LdNPV), occurring in spreading populations of an invasive forest defoliator, L. dispar (gypsy moth), in central Wisconsin. 3. Over 3 years, we found that host density was closely associated with the presence and prevalence of both pathogens. The fungal and viral pathogens differed in the sensitivity of their response as E. maimaiga was present in lower-density host population than LdNPV. 4. We examined the relationship between weather conditions and infection prevalence and found that activity of both the fungus and virus was strongly seasonally influenced by temperature and rainfall or temperature alone, respectively. 5. Purposeful releases of pathogens (median distances of study sites from release sites were 65.2 km for E. maimaiga and 25.6 km for LdNPV) were not associated with pathogen prevalence. 6. A generalist fly parasitoid, Compsilura concinnata, also killed L. dispar larvae collected from the study sites, and parasitism was greater when infection by pathogens was lower. 7. Our results demonstrated that although infection levels were low in newly established host populations, host-specific pathogens had already moved into host populations close behind advancing populations of an invasive host; thus, spreading hosts were released from these enemies for only a relatively short time. PMID- 21644979 TI - Differential expression and costs between maternally and paternally derived immune priming for offspring in an insect. AB - 1. When parasitized, both vertebrates and invertebrates can enhance the immune defence of their offspring, although this transfer of immunity is achieved by different mechanisms. In some insects, immune-challenged males can also initiate trans-generational immune priming (TGIP), but its expressions appear qualitatively different from the one induced by females similarly challenged. 2. The existence of male TGIP challenges the traditional view of the parental investment theory, which predicts that females should invest more into their progeny than males. However, sexual dimorphism in life-history strategies and the potential costs associated with TGIP may nevertheless lead to dissymmetric investment between males and females into the immune protection of the offspring. 3. Using the yellow mealworm beetle, Tenebrio molitor, we show that after parental exposure to a bacterial-like infection, maternal and paternal TGIP are associated with the enhancement of different immune effectors and different fitness costs in the offspring. While all the offspring produced by challenged mothers had enhanced immune defence, only those from early reproductive episodes were immune primed by challenged fathers. 4. Despite the fact that males and females may share a common interest in providing their offspring with an immune protection from the current pathogenic threat, they seem to have evolved different strategies concerning this investment. PMID- 21644980 TI - Complex interactions between components of individual prey specialization affect mechanisms of niche variation in a grasshopper-hunting wasp. AB - 1. Individual foraging behaviour defines the use of resources by a given population and its variation in different ways such as, for example, unpredictable interactions between taxon-biased and size-biased selection. Here we investigated how the environmental availability of prey and individual specialization, for both prey taxa and prey size, shape niche variation across generations in the grasshopper-hunting digger wasp Stizus continuus. 2. The population of S. continuus expressed selective predation, females mainly hunting species encountered on large bushes; diet changed across generations, due more to size increase in potential prey than to changes in the orthopteran community. 3. Individual females of both generations weakly overlapped the size and taxa of prey, and the niche width of the second generation increased for both prey size and taxa. 4. The greater variance in prey size in the environment accounted for the enlarged prey size niche of the second generation, but the load-lifting constraints of the wasps maintained individual prey size specialization constant. In contrast, the enlarged prey taxon niche paralleled a smaller overlap of diets between wasps in the second generation. 5. Increased niche width in the S. continuus population was thus achieved in two ways. Regarding prey size, all individuals shifted towards the use of the full set of available resources (parallel release). For prey taxa, according to the classical niche variation hypothesis, individuals diverged to minimize resource use overlap and perhaps intraspecific competition. These two mechanisms were observed for the first time simultaneously in a single predator population. PMID- 21644981 TI - Flexible components of functional responses. AB - 1. The functional response of predators describes the rate at which a predator consumes prey and is an important determinant of community dynamics. Despite the importance, most empirical studies have considered a limited number of models of functional response. In addition, the models often make strong assumptions about the pattern of predation processes, even though functional responses can potentially exhibit a wide variety of patterns. 2. In addition to the limited model consideration, model selections of functional response models cannot tease apart the components of predation (i.e. capture rate and handling time) when flexible traits are considered because it is always possible that many different combinations of the capture rate and handling time can lead to the same predation rate. 3. This study directly examined the capture rate and handling time of functional response in a mite community. To avoid the model selection problem, the searching and handling behaviour data were collected. The model selection was applied directly to these two components of predation data. Commonly used functional response models and models that allow for more flexible patterns were compared. 4. The results indicated that assumptions of the commonly used models were not supported by the data, and a flexible model was selected as the best model. These results suggest the need to consider a wider variety of predation patterns when characterizing a functional response. Without making a strong assumption (e.g. static handling time), model selections on functional response models cannot be used to make reliable inferences on the predation mechanisms. PMID- 21644982 TI - Lifetime fitness correlates of natal dispersal distance in a colonial bird. AB - 1. Obtaining empirical evidence of the consequences of dispersal distance on fitness is challenging in wild animals because long-term, unbiased data on reproduction, survival and movement are notoriously difficult to obtain. 2. Lifetime fitness correlates of natal dispersal distance were studied in an isolated population of the facultatively colonial lesser kestrel Falco naumanni (Fleischer) monitored during 8 years at north-eastern Spain, where most birds (83%) dispersed from their natal colony to settle at distances ranging from 112 m to 136.5 km. 3. Neither annual breeding success nor age at recruitment was affected by natal dispersal distance. However, a capture-mark-recapture analysis revealed that survival during the year following recruitment decreased exponentially with dispersal distance, with differences of up to 15% between philopatrics and long-distance dispersers. In subsequent years, it remained similar irrespective of the natal dispersal distance moved. These results did not seem to be biased by long-distance dispersers settling differentially in the periphery of the population (which could emigrate permanently and be considered dead in future occasions) or within-individual consistency in successive dispersal distances, so our results appear to reflect genuine survival differences between dispersal tactics. 4. Average lifetime fledgling production, average lifetime recruitment success and rate-sensitive individual fitness (lambda(ind)) also decreased with the distance from the natal to the first breeding colony, indicating that dispersal decisions early in life affecting immediate survival prospects may translate into long-term fitness costs. 5. Both survival and lifetime fitness models including continuous dispersal distances significantly improved the characterization of the effect on fitness compared with models considering dispersal as a discrete process (i.e. dispersal vs. philopatry at a colony level). 6. Long-distance dispersers were more likely to establish new colonies regardless of whether they recruited in the centre or the periphery of the population, revealing their important role in the colonization of unoccupied patches. Individuals experienced a higher probability of mortality in small and newly funded colonies, so lifetime fitness costs of dispersal seem to be explained by recruitment in sites where average quality is low because of high uncertainty in survival prospects. PMID- 21644983 TI - Editorial: the first cut is the deepest: why do the reported effects of treatments decline over trials? PMID- 21644984 TI - Randomized controlled trial of full and brief cognitive-behaviour therapy and wait-list for paediatric obsessive-compulsive disorder. AB - BACKGROUND: Reviews and practice guidelines for paediatric obsessive-compulsive disorder (OCD) recommend cognitive-behaviour therapy (CBT) as the psychological treatment of choice, but note that it has not been sufficiently evaluated for children and adolescents and that more randomized controlled trials are needed. The aim of this trial was to evaluate effectiveness and optimal delivery of CBT, emphasizing cognitive interventions. METHODS: A total of 96 children and adolescents with OCD were randomly allocated to the three conditions each of approximately 12 weeks duration: full CBT (average therapist contact: 12 sessions) and brief CBT (average contact: 5 sessions, with use of therapist guided workbooks), and wait-list/delayed treatment. The primary outcome measure was the child version of the semi-structured interviewer-based Yale-Brown Obsessive Compulsive Scale. CLINICAL TRIAL REGISTRATION: http://www.controlled trials.com/ISRCTN/; unique identifier: ISRCTN29092580. RESULTS: There was statistically significant symptomatic improvement in both treatment groups compared with the wait-list group, with no significant differences in outcomes between the two treatment groups. Controlled treatment effect sizes in intention to-treat analyses were 2.2 for full CBT and 1.6 for brief CBT. Improvements were maintained at follow-up an average of 14 weeks later. CONCLUSIONS: The findings demonstrate the benefits of CBT emphasizing cognitive interventions for children and adolescents with OCD and suggest that relatively lower therapist intensity delivery with use of therapist-guided workbooks is an efficient mode of delivery. PMID- 21644986 TI - "Grown-up" congenital heart disease and sudden death in a medical examiner's population. AB - Despite advances in the management of congenital heart disease (CHD), children with CHD who survive into adulthood are at increased risk of sudden death. Sudden death may also be the initial presentation of undiagnosed CHD in some adults. This retrospective descriptive study outlines the spectrum of CHD presenting as sudden death in adults in a medical examiner's population. Despite its rarity (0.2% of all cases investigated between 1991 and 2007), CHD remains an important cause of sudden cardiac death to be recognized at adult autopsy. Bicuspid aortic valve and anomalous coronary anomalies were the most common malformations, comprising 36.9% and 26.2% of cases, respectively. However, a wide spectrum of simple to complex malformations can be seen, with or without prior surgery, and over a wide age spectrum. Once solely a pediatric entity, CHD is now "grown-up" and will likely be diagnosed by forensic pathologists with increased frequency in the future. PMID- 21644985 TI - Stress-induced activation of the HPA axis predicts connectivity between subgenual cingulate and salience network during rest in adolescents. AB - BACKGROUND: Responses to stress vary greatly in young adolescents, and little is known about neural correlates of the stress response in youth. The purpose of this study was to examine whether variability in cortisol responsivity following a social stress test in young adolescents is associated with altered neural functional connectivity (FC) of the salience network (SN) measured during resting state functional magnetic resonance imaging (rs-fMRI). METHODS: Forty-nine typically developing young adolescents participated in a social stress test during which they contributed salivary cortisol samples. Following this, they underwent rs-fMRI scanning. We examined the association of FC of the SN [composed of anterior cingulate cortex and bilateral anterior insula regions] with cortisol responsivity. RESULTS: Greater cortisol responsivity was significantly positively correlated with higher FC between subgenual anterior cingulate cortex (Cg25) and the SN, controlling for participant age. There were no regions of the brain that showed an inverse relation. CONCLUSIONS: Brain systems that have been implicated in autonomic arousal and that influence subjective feeling states show altered FC associated with stress responsivity in early life. PMID- 21644987 TI - Algor mortis: an erroneous measurement following postmortem refrigeration. AB - Determination of the time of death is one goal of medicolegal death investigations. Algor mortis has been used as a measure of the postmortem interval (PMI). We prospectively recorded the core temperatures of 19 adult bodies entering our morgue cooler and at 3, 6, and 9 h of refrigeration. We then compared the cooling rate with the calculated body mass index (BMI). For each individual body, the rate of cooling was fairly linear with no evidence of a plateau. There was fair to moderate correlation between the BMI and the cooling rate: cooling rate = -0.052 (BMI) + 3.52. The probability of linearity in any given case was 36%. Variables affecting this correlation included the presence and the layers of clothing and if the clothing was wet. Our data confirm that algor mortis is of very limited utility in determining the PMI in bodies that have been refrigerated. PMID- 21644988 TI - Fatal intoxication because of trihexyphenidyl. AB - Trihexyphenidyl (THP) is an anticholinergic agent with forensic toxicological interest. We present a case of a 59-year-old woman with a history of paranoid disorder, who was found dead in the house where she lived alone. The autopsy findings revealed no marked pathological changes. Toxicological analysis based on gas chromatography-mass spectrometry (GC-MS) analysis revealed THP and its major metabolite (hydroxy-THP) in blood and urine, with THP concentrations of 0.053 and 0.560 mg/L, respectively. The blood and urine ethanol concentrations were low 0.096 and 0.100 g/L, respectively. Based on these results, we determined the cause of death to be THP poisoning. It is suggested that rare case of death associated with THP overdosage should be taken in conjunction with central nervous system depressants (benzodiazepines, ethanol) and/or with other pathological disorders. Thus, our case could not be supportive for this allegation. PMID- 21644989 TI - Occlusion of left and right coronary arteries and coronary sinus following blunt chest trauma. AB - Blunt chest trauma from rapid automobile airbag deployment causing coronary artery occlusion and myocardial infarction is a rare but potentially fatal condition. We present the case of a 37-year-old man who developed extensive anterior and inferior myocardial infarction because of occlusion of both left anterior and right coronary arteries following blunt injury to the chest in a car accident. The patient was scheduled for emergency coronary angiography but left and right coronary ostia were not cannulated because of thrombus formation probably. The patient died, and the autopsy revealed external compression by epicardial hematomas involving separately left and right coronary arteries and the coronary sinus without signs of coronary and/or aortic dissection. To our knowledge, this is the first case presenting occlusion of both coronary arteries secondary to blunt chest trauma causing acute myocardial infarction in a young man without signs of prior coronary artery disease. PMID- 21644990 TI - Field testing of collection cards for Cannabis sativa samples with a single hexanucleotide DNA marker. AB - The validity and feasibility of using DNA collection cards in the field for preservation and analysis of Cannabis sativa genotypes were investigated using a highly specific hexanucleotide marker. Collection cards were submitted to the National Marijuana Initiative, which selectively trained and managed the collection of specific types of samples from a variety of participating agencies. Samples collected at seizure sites included fresh marijuana leaf samples, dried "dispensary" samples, U.S. border seizures, and hashish. Using a standardized PCR kit with custom-labeled oligonucleotide primers specific to marijuana, collection cards produced eight genotypes and 13 different alleles, extremely low baselines, and no cross-reactivity with control plant species. Results were produced from all sample types with the exception of hashish. Plant DNA collection cards represent an easily implementable method for the genetic identification and relatedness of C. sativa street and grow site-seized samples with applications for databasing and market disruption. PMID- 21644991 TI - A comprehensive regression tree to estimate body weight from the skeleton. AB - The purpose of this research is to estimate actual body weight (in particular obesity) from the human skeleton. Known individuals (total n=187) were studied from the Bass Collection from the University of Tennessee, Knoxville. This research combines bone density, cross-sectional geometry of the femur and skeletal pathologies. Bone mineral density was calculated for the proximal femur. Three-dimensional bone surface models were created from computed tomographic scans. Cross-sectional geometry of the femur was calculated at five locations along the diaphysis. The pathologies analyzed were heel spurs, diffuse idiopathic skeletal hyperostosis (DISH), and tibial osteoarthritis. The best regression tree model included only four variables. The first split to estimate body weight was the minimum moment of inertia (I(y) ) at the distal femur, the second was midshaft width, then anteroposterior thickness at the proximal slice and the final split was the degree of DISH (SD 17.1-31.0 kg). The ability to estimate body weight from the skeleton is one more useful tool for the osteologist. PMID- 21644992 TI - A radiographic assessment of pediatric fracture healing and time since injury. AB - Past studies and pediatric bone physiology indicate that younger individuals may heal at a faster rate. Additionally, in adults upper limb fractures heal faster than lower limb fractures; this trend is expected for pediatric fractures. This study aims to evaluate and compare rates of fracture repair in children based on age and skeletal element. Six stages are used to describe the bone repair process in 294 radiographs of tibial and radial fractures from 107 infants and young children. Healing rates are examined using ANOVA and Welch's t-test with a 95% confidence interval. Results indicate that younger individuals spend less time at stage 1, suggesting a delay in the start of healing. Furthermore, forearm fractures heal faster than leg fractures at stages 2 and 3, suggesting a role in the osseous reaction of bone healing. The healing schedule presented may allow the timing of injuries to be estimated from radiographs. PMID- 21644993 TI - Complex suicide: an unusual case with six methods applied. AB - Complex suicides (CSs) are committed by using more than one method. They account for 1.5-5% of all suicides. We present a case of CSs of a 44-year-old man, found dead in the vicinity of his car, in a deserted frozen field. Police investigation excluded homicide, and no medical data confirmed mental illnesses. Autopsy revealed wrist cuts, neck cuts, acid burns in the GI tract, multiple stab wounds to the head by a screwdriver, and several uncertain signs of hypothermia. Toxicology analysis (gas chromatography-mass spectrometry) confirmed ingestion of insecticide. We concluded that stab wounds to the head were the cause of death, while external hemorrhage and hypothermia were contributing factors. This is the first case of CSs reviewed in the literature where six suicide methods were applied. This particular case is interesting because the victim used a screwdriver as a tool for inflicting stab wounds to the head, which is a rare suicidal method. PMID- 21644994 TI - Dopamine release mediated by the dopamine transporter, facts and consequences. AB - Spontaneous and/or stimulated neural activity of the nigrostriatal dopamine (DA) pathway makes amines run out from the neurons. This DA dynamic follows a rather complex path, running in or out the terminals, and flushing or diffusing into the extracellular space. The location of this leakage is not limited to the axon terminals; it also occurs from the cell bodies and dendrites. This molecular release mechanism was, for a long time, considered as being produced, in part, by the exocytosis of previously stored vesicles. The DA carrier protein (DAT, DA transporter) embedded in the DA cell membrane is known to clear previously released amines through an inward DA influx. The DAT also appears to be an active vector of amine release. Particular local conditions and the presence of numerous psychostimulant substances are able to trigger an outward efflux of DA through the DAT. This process, delivering slowly large amounts of amine could play a major regulatory role in extracellular DA homeostasis. PMID- 21644995 TI - Endogenous presynaptic nitric oxide supports an anterograde signaling in the central nervous system. AB - The source size and density determine the extent of nitric oxide (NO) diffusion which critically influences NO signaling. In the brain, NO released from postsynaptic somas following NMDA-mediated activation of neuronal nitric oxide synthase (nNOS) retrogradely affects smaller presynaptic targets. By contrast, in guinea pig trigeminal motor nucleus (TMN), NO is produced presynaptically by tiny and disperse nNOS-containing terminals that innervate large nNOS-negative motoneurons expressing the soluble guanylyl-cyclase (sGC); consequently, it is uncertain whether endogenous NO supports an anterograde signaling between pre motor terminals and postsynaptic trigeminal motoneurons. In retrogradely labeled motoneurons, we indirectly monitored NO using triazolofluorescein (DAF-2T) fluorescence, and evaluated sGC activity by confocal cGMP immunofluorescence. Multiple fibers stimulation enhanced NO content and cGMP immunofluorescence into numerous nNOS-negative motoneurons; NOS inhibitors prevented depolarization induced effects, whereas NO donors mimicked them. Enhance of cGMP immunofluorescence required extracellular Ca(2+), a nNOS-physiological activator, and was prevented by inhibiting sGC, silencing neuronal activity or impeding NO diffusion. In conclusion, NO released presynaptically from multiple cooperative tiny fibers attains concentrations sufficient to activate sGC in many motoneurons despite of the low source/target size ratio and source dispersion; thus, endogenous NO is an effective anterograde neuromodulator. By adjusting nNOS activation, presynaptic Ca(2+) might modulate the NO diffusion field in the TMN. PMID- 21644996 TI - Passive immunization targeting pathological phospho-tau protein in a mouse model reduces functional decline and clears tau aggregates from the brain. AB - Targeting hyperphosphorylated tau by immunotherapy is emerging as a promising approach to treat tauopathies such as Alzheimer's disease and frontotemporal dementia. We have previously reported that active tau immunization clears tau aggregates from the brain and attenuates or prevents functional impairments in two different tangle mouse models. Here, we assessed the efficacy of passive immunization with the PHF1 antibody, which targets a phospho-epitope within one of our active immunogens. Homozygous female tangle mice (JNPL3, 2-3 months) were injected intraperitoneally once per week with PHF1 or pooled mouse IgG (250 MUg/125 MUL; n = 10 per group) for a total of 13 injections. Their behavior was assessed at 5-6 months of age and brain tissue was subsequently harvested for analyses of treatment efficacy. The treated mice performed better than controls on the traverse beam task (p < 0.03), and had 58% less tau pathology in the dentate gyrus of the hippocampus (p = 0.02). As assessed by western blots, the antibody therapy reduced the levels of insoluble pathological tau by 14-27% (PHF1, p < 0.05; PHF1/total tau, p < 0.0001) and 34-45% (CP13 or CP13/total tau, p < 0.05). Levels of soluble tau and sarkosyl soluble tau were unchanged, compared with controls, as well as total tau levels in all the fractions. Plasma levels of PHF1 correlated inversely with tau pathology in the brainstem (p < 0.01), with a strong trend in the motor cortex (p < 0.06) as well as with insoluble total tau levels (p < 0.02), indicating that higher dose of antibodies may have a greater therapeutic effect. Significant correlation was also observed between performance on the traverse beam task and PHF1 immunoreactivity in the dentate gyrus (p < 0.05) as well as with insoluble PHF1/total tau ratio on western blots (p < 0.04). These results show that passive immunization with tau antibodies can decrease tau pathology and functional impairments in the JNPL3 model. Future studies will determine the feasibility of this approach with other monoclonals and in different tangle models in which thorough cognitive assessment can be performed. PMID- 21644998 TI - Ultrasound stimulation attenuates root resorption of rat replanted molars and impairs tumor necrosis factor-alpha signaling in vitro. AB - BACKGROUND AND OBJECTIVE: A therapeutic protocol to minimize root resorption induced by tooth replantation has not yet been universally established. In this context, noninvasive modality such as ultrasound therapy have been a focus of increased interest. This study aimed to evaluate the inhibitory effect of ultrasound therapy on root resorption of replanted rat molars. In addition, the study aimed to promote insights into the mechanism through which ultrasound mediates the metabolism of periodontal cells in vitro. MATERIAL AND METHODS: An experimental model of tooth replantation in rats, involving luxation and immediate replacement of the maxillary first molars, was used to assess the inhibitory effect of an ultrasound-therapy regimen (15 min of exposure to ultrasound, each day for 21 d) on root resorption. Moreover, the effect of ultrasound on osteoclastogenesis/cementoclastogenesis was examined in vitro using a mouse osteoblastic stromal cell line (ST2) and a mouse cementoblastic cell line (OCCM-30). RESULTS: The area of root resorption lacunae was statistically decreased (p < 0.01) in the ultrasound-treated sample. In addition, immunohistochemical staining, using murine TNF-alpha polyclonal antibody, failed to detect tumor necrosis factor-alpha (TNF-alpha) protein in the ultrasound treated sample compared with the control. An in vitro study showed that the lipopolysaccharide (LPS)-induced expression of Tnfalpha mRNA was significantly reduced by ultrasound therapy in both osteoblastic and cementoblastic cells. Moreover, the TNF-alpha-induced up-regulation of Rankl mRNA was also inhibited by ultrasound. CONCLUSION: Ultrasound may contribute to the reduction of the trauma induced inflammatory reaction through impairment of the TNF-alpha signaling pathway. It is therefore suggested that ultrasound shows potential as a therapeutic tool to optimize the regenerative potential of periodontal tissues on replanted teeth. PMID- 21644997 TI - Advances in understanding the peptide neurotransmitter NAAG and appearance of a new member of the NAAG neuropeptide family. AB - A substantial body of data was reported between 1984 and 2000 demonstrating that the neuropeptide N-acetylaspartylglutamate (NAAG) not only functions as a neurotransmitter but also is the third most prevalent transmitter in the mammalian nervous system behind glutamate and GABA. By 2005, this conclusion was validated further through a series of studies in vivo and in vitro. The primary enzyme responsible for the inactivation of NAAG following its synaptic release had been cloned, characterized and knocked out. Potent inhibitors of this enzyme were developed and their efficacy has been extensively studied in a series of animal models of clinical conditions, including stroke, peripheral neuropathy, traumatic brain injury, inflammatory and neuropathic pain, cocaine addiction, and schizophrenia. Considerable progress also has been made in defining further the mechanism of action of these peptidase inhibitors in elevating synaptic levels of NAAG with the consequent inhibition of transmitter release via the activation of pre-synaptic metabotropic glutamate receptor 3 by this peptide. Very recent discoveries include identification of two different nervous system enzymes that mediate the synthesis of NAAG from N-acetylaspartate and glutamate and the finding that one of these enzymes also mediates the synthesis of a second member of the NAAG family of neuropeptides, N-acetylaspartylglutamylglutamate. PMID- 21645000 TI - Cloning and characterization of barley caryopsis FCA. AB - The RNA binding protein, flowering control locus A, (FCA) regulates flowering in rice and Arabidopsis. FCA interacts with FY to auto-regulate its own transcripts as well as to control flowering by downregulating flowering locus C (FLC). We report the cloning and characterization of the gamma (gamma) isoform of FCA from barley (Hordeum vulgare cv. McLeod). The deduced protein contained two RNA recognition motifs (RRMs), a glycine-rich region at the N-terminal end, a polyglutamine region immediately downstream of a WW domain. Barley FCA had greater protein sequence homology to wheat and rice FCA than to its Arabidopsis homolog. In developing barley embryos, FCA transcripts could be detected from 2 days after pollination (DAP) up to 40 DAP. FCA transcript levels in mature barley embryo were more abundant in non-germinated than in germinated seeds, with the levels declining as germination progressed. ABA inhibition of germination inhibited the decline of barley embryo FCA. Transient co-expression of FCA or a truncated FCA (lacking RRM) with maize VP1 promoter or wheat Em gene promoter in barley aleurone protoplasts led to increased VP1 and Em gene promoter activity. Barley FCA or truncated FCA localized in the nucleus suggested its role in gene regulation. PMID- 21644999 TI - Characterization of human-type monoclonal antibodies against reduced form of hemin binding protein 35 from Porphyromonas gingivalis. AB - BACKGROUND AND OBJECTIVE: The gram-negative anaerobe Porphyromonas gingivalis has been implicated as an important pathogen in the development of adult periodontitis, and its colonization of subgingival sites is critical in the pathogenic process. We previously identified a 35 kDa surface protein (hemin binding protein 35; HBP35) from P. gingivalis that exhibited coaggregation activity, while additional analysis suggested that this protein possessed an ability to bind heme molecules. For development of passive immunotherapy for periodontal diseases, human-type monoclonal antibodies have been prepared using HBP35 as an antigen in TransChromo mice. In the present study, we focused on a single antibody, TCmAb-h13, which is known to inhibit heme binding to recombinant HBP35. The aim of our investigation was to clarify the redox-related function of HBP35 and consider the benefits of human-type monoclonal antibodies. MATERIAL AND METHODS: To examine the antigen recognition capability of TCmAbs with immunoblotting and Biacore techniques, we used the native form as well as several Cys-to-Ser variants of recombinant HBP35. RESULTS: We found that the redox state of recombinant HBP35 was dependent on two Cys residues, (48) C and (51) C, in the thioredoxin active center (WCGxCx). Furthermore, TCmAb-h13 recognized the reduced forms of recombinant HBP35, indicating its inhibitory effect on P. gingivalis growth. CONCLUSION: Hemin binding protein 35 appears to be an important molecule involved in recognition of the redox state of environmental conditions. In addition, TCmAb-h13 had an inhibitory effect on heme binding to recombinant HBP35, thereby interfering with P. gingivalis growth. PMID- 21645002 TI - Invasive Eupatorium adenophorum has ecophysiological advantages over native congeners but similar responses to CO(2) enrichment. AB - Both global change and biological invasions threaten biodiversity worldwide. However, their interactions and related mechanisms are still not well elucidated. To elucidate potential traits contributing to invasiveness and whether ongoing increase in CO(2) aggravates invasions, noxious invasive Eupatorium adenophorum and native E. japonicum and E. chinensis were compared under ambient and doubled atmospheric CO(2) concentrations in terms of growth, biomass allocation, morphology, and physiology. The invader had consistently higher leaf mass fraction and specific leaf area than the natives, contributing to a higher leaf area ratio, and therefore to faster growth and invasiveness. The higher leaf mass fraction of the invader was associated with lower total root mass fraction. The invader allocated a higher fraction of leaf nitrogen (N) to photosynthesis, contributing to higher area-based N content in photosynthetic apparatus, photosynthetic rate, nitrogen- and water-use efficiencies, and invasiveness. CO(2) enrichment increased growth of all studied plants by increasing actual photosynthesis, although it decreased photosynthetic capacities due to decreased area-based leaf and photosynthetic N contents. Responses of the invasive and native plants to elevated CO(2) were not significantly different, indicating that the ongoing increase in CO(2) may not exacerbate biological invasions, inconsistent with the prevailing results in references. The difference may be associated with the fact that almost all previous studies compared phylogenetically unrelated invasive and native plants. More comparative studies of sympatric, related invasive and native plants are needed to elucidate whether CO(2) enrichment facilitates invasions. PMID- 21645001 TI - Variations of DNA methylation in Eucalyptus urophylla*Eucalyptus grandis shoot tips and apical meristems of different physiological ages. AB - Global DNA methylation was assessed by high-performance liquid chromatography (HPLC) for the first time in Eucalyptus urophylla*Eucalyptus grandis shoot tips comparing three outdoor and one in vitro sources of related genotypes differing in their physiological age. The DNA methylation levels found were consistent with those reported for other Angiosperms using the same HPLC technology. Notwithstanding noticeable time-related fluctuations within each source of plant material, methylation rate was overall higher for the mature clone (13.7%) than for the rejuvenated line of the same clone (12.6%) and for the juvenile offspring seedlings (11.8%). The in vitro microshoots of the mature clone were less methylated (11.3%) than the other outdoor origins, but the difference with the juvenile seedlings was not significant. Immunofluorescence investigations on shoot apices established that the mature source could be distinguished from the rejuvenated and juvenile origins by a higher density of cells with methylated nuclei in leaf primordia. Shoot apical meristems (SAMs) from the mature clone also showed a greater proportion and more methylated cells than SAMs from the rejuvenated and juvenile origins. The nuclei of these latter were characterized by fewer and more dispersed labeled spots than for the mature source. Our findings establish that physiological ageing induced quantitative and qualitative variations of DNA methylation at shoot tip, SAM and even cellular levels. Overall this DNA methylation increased with maturation and conversely decreased with rejuvenation to reach the lower scores and to show the immunolabeling patterns that characterized juvenile material nuclei. PMID- 21645003 TI - Slowly developing drought stress increases photosynthetic acclimation of Catharanthus roseus. AB - Our understanding of plant responses to drought has improved over the decades. However, the importance of the rate of drought imposition on the response is still poorly understood. To test the importance of the rate at which drought stress develops, whole-plant photosynthesis (P(net) ), respiration (R(dark) ), daily carbon gain (DCG), daily evapotranspiration (DET) and water use efficiency (WUE) of vinca (Catharanthus roseus), subjected to different drought imposition rates, were investigated. We controlled the rate at which the substrate dried out with an automated irrigation system that allowed pot weight to decrease gradually throughout the drying period. Fast, intermediate and slow drying treatments reached their final pot weight [500 g, substrate water content (theta) ~ 0.10 m3 m(-3) ] after 3.1, 6.6 and 10 days, respectively. Although all drying treatments decreased P(net) and R(dark) , slow drying reduced P(net) and R(dark) less than fast drying. At a theta < 0.10 m3 m(-3) , DCG and DET in the slow drying treatment were reduced by ~50%, whereas DCG and DET in the fast drying treatment were reduced by 85 and 70% at a theta of 0.16 m(3) m(-3) . Plants exposed to slow drought imposition maintained a high WUE, even at theta < 0.10 m3 m(-3) . Overall, physiological responses to low theta were less severe in plants subjected to slow drying as compared with fast drying, even though the final theta was lower for plants exposed to slow drying. This suggests that the rate at which drought stress develops has important implications for the level of acclimation that occurs. PMID- 21645004 TI - Quantitation of coated platelet potential during collection, storage, and transfusion of apheresis platelets. AB - BACKGROUND: Coated platelets (PLTs), a subpopulation of PLTs observed upon dual agonist stimulation with collagen and thrombin, are known to retain several procoagulant alpha-granule proteins on their surface. By formation of a highly active membrane-bound prothrombinase complex, these PLTs represent an important step in the coagulation cascade as a consequence of their ability to generate thrombin at the site of vascular injury. Various clinical observations suggest that higher levels of coated PLTs are associated with thrombosis while a deficiency of coated PLTs results in a bleeding diathesis. Current quality control guidelines for in vitro PLT storage measure PLT viability but no routine evaluation of the hemostatic function of stored PLTs and particularly no estimation of coated PLT potential is performed. Our primary objective was to evaluate if the process of apheresis and storage of PLT units alters the levels of coated PLTs. In addition, we sought to determine how transfusion of stored PLTs into patients with thrombocytopenia affects the patient's coated PLT levels. STUDY DESIGN AND METHODS: Coated PLT levels were analyzed in 13 voluntary PLT donors before donation, in the fresh apheresis product (Trima, CaridianBCT) and in the stored apheresis product just before transfusion. In addition, 10 patients with thrombocytopenia were analyzed for coated PLTs before and after transfusion of a stored PLT product. RESULTS: Coated PLT levels were significantly decreased after the process of apheresis (17% relative decline; p < 0.01) and with prolonged storage (1 to 5 days; 53% relative decline; p < 0.001). Transfusion of stored PLT units did not result in significant increment of coated PLT levels in patients with thrombocytopenia as expected considering the low level of coated PLTs in stored PLT units. Furthermore, there was no suggestion of regeneration of coated PLT potential upon reinfusion. CONCLUSIONS: Isolation and storage of apheresis PLTs by standard blood bank procedures results in a significant decline in coated PLT potential. Reinfusion of stored apheresis PLTs into patients with thrombocytopenia resulted in a predictable change in coated PLT potential with no suggestion of regeneration of lost coated PLT potential. PMID- 21645005 TI - Transfusion of fresh murine red blood cells reverses adverse effects of older stored red blood cells. AB - BACKGROUND: Although a subset of recent studies have suggested that red blood cell (RBC) storage length is associated with adverse patient outcomes, others have shown no such relationship. Adults may be transfused with RBC units of different storage lengths, and existing studies do not take into consideration that fresh RBCs may alter responses to concurrently transfused stored RBCs. To test this possibility, we utilized a murine model and investigated transfusion outcomes of fresh, stored, or fresh-plus-stored RBCs. STUDY DESIGN AND METHODS: Fresh, 14-day-stored or fresh plus 14-day-stored leukoreduced RBCs from HOD transgenic donors (with RBC-specific expression of hen egg lysozyme, ovalbumin, and human Duffy(b)) were transfused into naive C57BL/6 recipients. Serum cytokines and anti-HOD alloimmunization were evaluated after transfusion. RESULTS: In six of six experiments (n = 90 mice total), a proinflammatory serum cytokine storm of interleukin-6, keratinocyte-derived chemokine/CXCL1, and monocyte chemoattractant protein-1 was observed in transfusion recipients of stored but not fresh RBCs, along with high degrees of anti-HOD alloimmunization. However, concurrent transfusion of fresh HOD RBCs along with stored HOD RBCs significantly decreased these adverse outcomes (p < 0.05). CONCLUSIONS: These results are consistent with fresh murine HOD RBCs losing protective properties during storage, and introduce a previously unrecognized variable in RBC storage studies. If translatable to humans, uniform "old blood" groups may be needed in future clinical studies to more accurately investigate the biologic effects of older RBC units. PMID- 21645006 TI - Estimation of variant Creutzfeldt-Jakob disease infectivity titers in human blood. AB - BACKGROUND: Blood of individuals with variant Creutzfeldt-Jakob disease (vCJD) is infectious but the titer is unknown. Current estimates of possible vCJD infectivity titers in blood have largely relied on an assumption that the titers of vCJD agent in human blood are likely to be similar to those in blood of rodents infected with model transmissible spongiform encephalopathy agents, assayed by intracerebral inoculations of rodents of the same species. STUDY DESIGN AND METHODS: We analyzed published descriptions of experimental transfusion-transmitted (TT) bovine spongiform encephalopathy and scrapie in sheep and reports of TTvCJD in humans, applying statistical approaches to estimate the probable number of intravenous infectious doses (ID(iv) ) per unit of transfused blood (ID(iv) /unit). For humans, ID(iv) /unit of nonleukoreduced red blood cells (NLR-RBCs) were estimated by two statistical models. RESULTS: Sheep blood collected at or near onset of clinical illness contained a mean of 0.80 ID(iv) /unit. Estimates of infectivity in NLR-RBCs from donors incubating vCJD indicated a probable mean infectivity of 0.29 ID(iv) /unit (Model 1) and 0.75 ID(iv) /unit (Model 2). The analysis predicted a mean of 21 vCJD-infected recipients expected in a cohort transfused with vCJD-implicated NLR-RBCs in the United Kingdom. CONCLUSION: Our analysis suggested that, while less than one ID(iv ) is likely to be present in a given unit of NLR-RBCs collected from a donor incubating vCJD, there is a high probability of TT infection among recipients of vCJD-implicated blood components. The analysis supports continuing measures currently recommended to reduce the risk of TTvCJD. PMID- 21645007 TI - Postoperative complications associated with transfusion of platelets and plasma in cardiac surgery. AB - BACKGROUND: Studies in cardiac surgery have reported increased postoperative morbidity and mortality after allogeneic red blood cell (RBC) transfusions. Whether platelet (PLT) and/or plasma transfusions are a marker for more concomitant RBC transfusions or are independently associated with complications after cardiac surgery is unknown. STUDY DESIGN AND METHODS: Data from two randomized controlled studies were combined to analyze the effects of PLT and/or plasma transfusions on postoperative infections, length of stay in the intensive care unit (ICU), all-cause mortality, and mortality in the presence or absence of infections in the postoperative period. RESULTS: After adjusting for confounding factors, plasma units and not RBC transfusions were associated with all-cause mortality. White blood cell (WBC)-containing RBC transfusions and PLT transfusions were associated with mortality occurring in the presence of or after infections. The number of (WBC-containing) RBC transfusions was also significantly associated with postoperative infections and with ICU stay for 4 or more days. CONCLUSION: Although it is difficult to separate the effects of blood components, we found that in cardiac surgery, perioperative plasma transfusions are independently associated with all-cause mortality. WBC-containing RBC transfusions and PLT transfusions are independently associated with mortality in the presence of infections in the postoperative period. Future transfusion studies in cardiac surgery should concomitantly consider the possible adverse effects of all the various transfused blood components. PMID- 21645008 TI - Physiologic strategies to prevent fainting responses during or after whole blood donation. AB - Vasovagal syncope (VVS) is a consistent, but infrequent (0.1%-0.3%) complication of volunteer, whole blood donation. Given the large number of blood donations, a significant number of donors is involved. Syncope occasionally leads to injury. Recent rigorous data collection and analysis have led to the association of a small number of donor and donation factors with the risk of syncope. An analysis of the time course of syncope reactions among approximately 500,000 whole blood donors suggests that there are three distinct periods of risk for vasovagal reactions before, during, and after phlebotomy. This review examines the physiologic mechanisms that contribute to these periods of increased risk including the direct effects of removal of approximately 500 mL of whole blood, the psychological stress of instrumentation and giving blood (i.e., fear of needles, pain, and the sight of blood), and the orthostatic effects superimposed on a hypovolemic state after the donation. Specifically, we describe interventions that have been useful in controlling VVS in patients with fainting syndromes and we examine the potential of these interventions in the blood donation context, based on the physiologic principles involved. Finally, we propose an intervention (dietary replacement of salt lost with blood donation) that has not been applied in transfusion medicine previously but which has the potential to reduce risk. PMID- 21645009 TI - Transmission of cytomegalovirus (CMV) infection by leukoreduced blood products not tested for CMV antibodies: a single-center prospective study in high-risk patients undergoing allogeneic hematopoietic stem cell transplantation (CME). AB - BACKGROUND: Measures to prevent transfusion-transmitted cytomegalovirus (TT-CMV) infection after hematopoietic stem cell transplantation (HSCT) include transfusion of CMV antibody-negative blood units and/or transfusion of leukoreduced cellular blood products. We assessed the incidence of TT-CMV in CMV seronegative patients receiving CMV-seronegative HSC transplants, who were transfused with leukoreduced cellular blood products not tested for anti-CMV. STUDY DESIGN AND METHODS: In a prospective observational study between 1999 and 2009, all HSCT patients received leukoreduced cellular blood products not tested for anti-CMV. Patients were screened for CMV serostatus and CMV-negative recipients of CMV-negative transplants were systematically monitored for TT-CMV clinically and by CMV nucleic acid testing. Anti-CMV antibodies (immunoglobulin [Ig]G and IgM) were assessed after three time intervals (Interval 1, study inclusion to Day +30 after HSCT; Interval 2, Day +30-Day +100; Interval 3, after Day +100). RESULTS: Among 142 patients treated with allogeneic HSCT, 23 CMV negative donor-patient pairs were identified. These 23 patients received 1847 blood products from 3180 donors. All patients remained negative for CMV DNA and none developed CMV-associated clinical complications. This results in a risk for TT-CMV per donor exposure of 0% (95% confidence interval, 0.0%-0.12%). However, 17 of 23 patients seroconverted for anti-CMV IgG, but none for anti-CMV IgM. CMV IgG seroconverters received significantly more transfusions per week than nonconverters. CONCLUSION: The risk of TT-CMV is low in high-risk CMV(neg/neg) HSCT patients transfused with leukoreduced blood products not tested for anti CMV. The cause of anti-CMV IgG seroconversion is most likely passive antibody transmission by blood products. PMID- 21645010 TI - Can anti-IgE therapy prevent airway remodeling in allergic asthma? AB - Airway remodeling is a central feature of asthma. It is exemplified by thickening of the lamina reticularis and structural changes to the epithelium, submucosa, smooth muscle, and vasculature of the airway wall. Airway remodeling may result from persistent airway inflammation. Immunoglobulin E (IgE) is an important mediator of allergic reactions and has a central role in airway inflammation and asthma-related symptoms. Anti-IgE therapies (such as omalizumab) have the potential to block an early step in the allergic cascade and therefore have the potential to reduce airway remodeling. The reduction in free IgE levels following anti-IgE therapy leads to reductions in high-affinity IgE receptor (FcepsilonRI) expression on mast cells, basophils, and dendritic cells. This combined effect results in attenuation of several markers of inflammation, including peripheral and bronchial tissue eosinophilia and levels of granulocyte macrophage colony stimulating factor, interleukin (IL)-2, IL-4, IL-5, and IL-13. Considering the previously demonstrated anti-inflammatory effects of anti-IgE therapy, along with results from a small study showing continued benefit after discontinuation of long-term treatment, a larger study to assess its effect on markers of airway remodeling is underway. PMID- 21645011 TI - IgG4 antibodies against rodents in laboratory animal workers. PMID- 21645012 TI - Molecular cloning and characterization of Fag t 2: a 16-kDa major allergen from Tartary buckwheat seeds. PMID- 21645013 TI - Drug-induced lichenoid reaction after kyphoplasty. PMID- 21645014 TI - The in vitro diagnosis of drug allergy: status and perspectives. AB - Adverse drug reactions (ADR) can result from immune-mediated (drug allergy) and nonimmune-mediated mechanisms. In both types of reaction, conclusive diagnosis and appropriate management remain major problems in daily clinical practice. This review summarizes the potentials and shortcomings of the currently available in vitro tests in the diagnosis of immediate (mostly IgE mediated) and nonimmediate (mostly T-cell mediated) drug allergy, particularly quantification of specific IgE, flow-assisted analysis of in vitro activated lymphocytes and basophils and the enzyme-linked immunosorbent spot. PMID- 21645015 TI - ABCG2 transporter: therapeutic and physiologic implications in veterinary species. AB - Drug transporters significantly influence drug pharmacokinetics and pharmacodynamics. While P-glycoprotein, the product of the MDR1 (ABCB1) gene, is the most well-characterized ABC transporter, the pharmacological importance of a related transporter, ABCG2, is starting to be realized in veterinary medicine. Based primarily on human and rodent studies, a number of clinically relevant, structurally and functionally unrelated drugs are substrates for ABCG2. ABCG2 is expressed by a variety of normal tissues including the intestines, renal tubular cells, brain and retinal capillary endothelial cells, biliary canalicular cells, and others, where it functions to actively extrude substrate drugs. In this capacity, ABCG2 limits oral absorption of substrate drugs and restricts their distribution to privileged sites such as the brain and retina. ABCG2 is also expressed by tumor cells where it functions to limit the intracellular accumulation of cytotoxic agents, contributing to multidrug resistance. Several ABCG2 polymorphisms have been described in human patients, some of which result in altered drug disposition, increasing susceptibility to adverse drug reactions. Additionally, ABCG2 polymorphisms in humans have been associated with disease states such as gout. Feline ABCG2 has recently been demonstrated to have several amino acid differences at conserved sites compared with 10 other mammalian species. These amino acid differences adversely affect transport function of feline ABCG2 relative to that of human ABCG2. Furthermore, these differences appear to be responsible for fluoroquinolone-induced retinal toxicity in cats and may play a role in acetaminophen toxicity as well. Studies in rodents and sheep have determined that ABCG2 expressed in mammary tissue is responsible for the secretion of many compounds (both therapeutic and toxic) into milk. Finally, data in rodent models suggest that ABCG2 may play an important role in regulating a number of physiologic pathways involved in protecting erythrocytes from oxidative damage. PMID- 21645016 TI - Metabolic regulation of antibiotic resistance. AB - It is generally assumed that antibiotics and resistance determinants are the task forces of a biological warfare in which each resistance determinant counteracts the activity of a specific antibiotic. According to this view, antibiotic resistance might be considered as a specific response to an injury, not necessarily linked to bacterial metabolism, except for the burden that the acquisition of resistance might impose on the bacteria (fitness costs). Nevertheless, it is known that changes in bacterial metabolism, such as those associated with dormancy or biofilm formation, modulate bacterial susceptibility to antibiotics (phenotypic resistance), indicating that there exists a linkage between bacterial metabolism and antibiotic resistance. The analyses of the intrinsic resistomes of bacterial pathogens also demonstrate that the building up of intrinsic resistance requires the concerted action of many elements, several of which play a relevant role in the bacterial metabolism. In this article, we will review the current knowledge on the linkage between bacterial metabolism and antibiotic resistance and will discuss the role of global metabolic regulators such as Crc in bacterial susceptibility to antibiotics. Given that growing into the human host requires a metabolic adaptation, we will discuss whether this adaptation might trigger resistance even in the absence of selective pressure by antibiotics. PMID- 21645017 TI - Selection and identification of autochthonous yeasts in Slovakian wine samples using a rapid and reliable three-step approach. AB - AIMS: The investigation of yeast microflora during the must fermentation of two wine varieties (Frankovka modra - Blaufrankisch and Veltlinske zelene - Gruner Veltliner) from two consecutive vintages was performed using a three-step approach. METHODS AND RESULTS: The investigation strategy consisted of the combination of yeast cultivation, selection of the isolated yeasts based on the amplification of internal transcribed spacer 2 using a fluorescence-labelled primer (f-ITS-PCR) and a final identification step based on amplification and sequencing of the ITS1-5.8S rDNA-ITS2 region of the selected yeasts. By this three-step approach, it was possible to screen 433 yeasts isolates that belonged to 13 different species. CONCLUSIONS: The f-ITS-PCR allowed the unambiguous differentiation of all isolated yeast species that produced their typical f-ITS PCR profile. SIGNIFICANCE AND IMPACT OF THE STUDY: This is one of few reports that treat the yeast diversity in Slovakian wines and in two varieties largely cultivated in Central Europe. The three-step approach permitted the rapid and reliable identification of isolated yeasts. The f-ITS-PCR with its good discrimination power can represent a suitable molecular tool for the selection of yeast members recovered from food or other environments. PMID- 21645018 TI - Adalimumab sustains steroid-free remission after 3 years of therapy for Crohn's disease. AB - BACKGROUND: Treatments that achieve sustainable steroid-free clinical remission in Crohn's disease are needed; however, long-term steroid-sparing efficacy data are limited. AIM: To evaluate steroid-sparing efficacy and the impact of steroid discontinuation on adverse events during treatment of Crohn's disease with adalimumab in the phase III randomised, double-blind 1-year CHARM trial and for an additional 2 years in its open-label extension ADHERE. METHODS: Steroid-free remission and response and steroid-sparing (>=50% steroid dose reduction) remission rates were evaluated over 3 years in patients who were taking corticosteroids at CHARM baseline. RESULTS: Of 778 patients randomised in CHARM (including those who did not achieve clinical response to open-label induction therapy), 313 patients (40%) were on corticosteroids at baseline. In the 206 patients randomised to adalimumab, rates of steroid-free remission at 1 year and 3 years were 26% and 23% respectively; corresponding rates were 29% and 25% for steroid-sparing remission and 32% and 28% for steroid-free response. Although the incidence of serious infections with adalimumab treatment during CHARM was higher in patients taking steroids at baseline than those who were not, the rates of overall adverse events, serious infections and opportunistic infections were lower in patients who were able to discontinue corticosteroids than those who remained on steroids. CONCLUSION: Adalimumab therapy resulted in modest but clinically meaningful rates of steroid-free remission, sustained over 3 years of treatment, in a heavily pretreated population of patients with Crohn's disease receiving steroids at the start of therapy (http://www.clinicaltrials.gov number: NCT00077779). PMID- 21645019 TI - Delayed response of human melanopsin retinal ganglion cells on the pupillary light reflex. AB - PURPOSE: A recent study has shown that retinal ganglion cells containing the photopigment melanopsin, which are intrinsically photosensitive in primates, project to the pupillary control centre in the pretectum. The aim of this study was to investigate how melanopsin retinal ganglion cells (mRGCs) contribute to the pupillary pathway. METHODS: We designed and built a novel multi-primary stimulation system to control stimulation of the three cone types and mRGCs independently in the human eye. We measured the latency and amplitude of transient pupillary responses to three types of test stimuli modulating excitations of mRGCs and cones (mRGC, luminance and the light flux stimuli). RESULTS: It was found that the transient pupillary response to mRGC stimuli has a longer latency than that to luminance and the light flux stimuli when an onset of sinusoidal stimulus was used. CONCLUSIONS: The results indicate that we successfully demonstrated the pupillary response to mRGCs under conditions where mRGCs are isolated in humans. Furthermore, the data confirm that the delayed response disappeared when the stimulus is presented as a square-wave pulse and not weighted by a sinusoid. The similarity of time courses for the earlier phase of pupillary responses to all stimuli suggested that these transient pupillary responses were driven by a single mechanism, which is perhaps associated with cone-mediated signals. PMID- 21645020 TI - Unacylated ghrelin is associated with changes in insulin sensitivity and lipid profile during an exercise intervention. AB - OBJECTIVE: Ghrelin has been implicated in energy homeostasis, body weight regulation and glucose metabolism. Level of unacylated ghrelin (UAG), but not acylated ghrelin (AG), has been suggested to increase during long-term exercise. However, the association of the level of UAG with exercise-induced changes of insulin sensitivity and lipid metabolism has not been previously investigated. We hypothesized that an increase in UAG level in response to a long-term exercise programme improves insulin sensitivity and associated lipid profile, independently of weight loss. DESIGN, PATIENTS AND MEASUREMENTS: A prospective study of 552 young men (mean age 19.3 and range 19-28 years) undergoing military service with structured 6-month exercise training programme. Exercise performance, clinical and biochemical measurements were obtained at baseline and follow-up. Association between UAG level and fasting glucose, insulin, insulin sensitivity and lipid levels were evaluated. RESULTS: An overall increase in the level of UAG was observed during the 6-month follow-up (P < 0.001), which was largest among those with weight loss >= 2.5% or among those whose reduction in waist circumference was largest (P = 0.007 and P < 0.001, respectively). A change in UAG level correlated inversely with a change in fasting glucose and insulin levels, HOMA-IR, total cholesterol and total triglyceride levels (P < 0.001 for all). The association between change in the UAG level and the change in insulin sensitivity was independent of weight loss or reduction in waist circumference. CONCLUSIONS: Increase in UAG level was associated with improved insulin sensitivity via mechanisms independent of weight loss during an intensive, long term exercise intervention in young healthy men. PMID- 21645021 TI - Recurrence of hyperprolactinaemia following discontinuation of dopamine agonist therapy in patients with prolactinoma occurs commonly especially in macroprolactinoma. AB - CONTEXT: The optimal duration of dopamine agonist (DA) therapy in prolactinoma is unknown. There are concerns that despite low recurrence rates in highly selected groups, high recurrence rates after DA withdrawal may occur in routine practice. OBJECTIVE: To explore recurrence of hyperprolactinaemia and predictive factors following DA withdrawal in patients with microprolactinoma and macroprolactinoma. DESIGN: A retrospective study on adult patients with confirmed prolactinoma attending the Oxford Endocrine Department. PATIENTS AND MEASUREMENTS: We identified patients with macroprolactinoma (n = 15) and microprolactinoma (n = 45) treated with DA therapy for >3 years, with a trial off DA therapy. None had other treatments. Measurements included recurrence of hyperprolactinaemia following DA withdrawal, tumour size (macroprolactinomas), duration of DA therapy, prolactin levels (baseline, during DA therapy, recurrence) and time to recurrence. Data were reported as mean (range). RESULTS: During DA therapy, prolactin levels suppressed to normal range in all patients with macroprolactinoma and microprolactinoma, and most macroprolactinomas (n = 14) had substantial tumour shrinkage. Hyperprolactinaemia recurred in 93% of macroprolactinomas (n = 14) at 8.8 months (3-36) and 64% of microprolactinomas (n = 29) at 4.8 months (3-12). Duration of DA therapy was 7.5 years (4-15) for macroprolactinomas and 4.1 years (3-10) for microprolactinomas. Prolactin levels during DA therapy were 144 mU/l (7-336) for macroprolactinomas and 278 mU/l (30 629) for microprolactinomas. For microprolactinomas, prolactin levels during DA therapy were less suppressed in those with recurrence than in those without recurrence (P < 0.05). CONCLUSIONS: In routine practice, hyperprolactinaemia recurs early in most macroprolactinomas (93%) and microprolactinomas (64%) following DA therapy discontinuation. For most macroprolactinomas, cessation of DA cannot be recommended even after 7 years of therapy. PMID- 21645022 TI - Early detection of isolated left ventricular diastolic dysfunction in high-risk differentiated thyroid carcinoma patients on TSH-suppressive therapy. AB - OBJECTIVE: L-Thyroxine-suppressive therapy benefits high-risk differentiated thyroid cancer patients by decreasing recurrence rates and cancer-related mortality. However, fully suppressed serum thyroid-stimulating hormone (TSH) implies a state of subclinical hyperthyroidism (SCH) with associated adverse cardiac effects. Because left ventricular (LV) diastolic dysfunction may be the first manifestation of more severe LV failure, and to balance the risks from thyroid cancer recurrence with risks of cardiac failure, the purpose of this study was to analyse new parameters of LV function in asymptomatic patients with exogenous SCH. DESIGN: Case-control study with 24 patients on TSH-suppressive therapy of short duration (<= 4 years) after thyroid ablative therapy for differentiated thyroid carcinoma and 20 age- and sex-matched subjects. MEASUREMENTS: LV function [LV global strain and strain rate (SR) curves] was assessed by speckle tracking imaging echocardiography in each subject. RESULTS: Patients and controls do not differ in body mass index, systolic blood pressure and heart rate. No significant differences were observed in LV morphology (LV mass and relative wall thickness), cardiac output and parameters of LV systolic function between patients on suppressive therapy and controls. When compared with controls, patients with exogenous SCH had a significantly impaired longitudinal protodiastolic strain, SR and strain diastolic index but preserved radial strain and SR function. CONCLUSIONS: In subjects with SCH at the early phase of TSH suppressive therapy, evidence of isolated longitudinal LV diastolic dysfunction was observed, despite a normal LV morphology. Further prospective studies to clarify the prognosis of picking-up early diastolic dysfunction in asymptomatic patients are needed before serial measurements could be recommended. PMID- 21645023 TI - Hyperinsulinemia acutely increases serum macrophage inhibitory cytokine-1 concentration in anorexia nervosa and obesity. AB - CONTEXT: Macrophage inhibitory cytokine-1 (MIC-1) plays a role in the regulation of cellular responses to stress signals and inflammation. MIC-1 has also been implicated in mediation of tumour-induced anorexia and weight loss. Increased serum concentrations of MIC-1 were found in patients with anorexia nervosa (AN), obesity and type 2 diabetes. OBJECTIVE: To estimate serum MIC-1 concentration in women with AN and obese women, its regulation by hyperinsulinemia and relationship with insulin sensitivity. PATIENTS: We examined 20 women with AN, 28 healthy normal-weight female controls and 28 obese women. MEASUREMENTS: Serum MIC 1 concentration was measured in the fasting state and after 2-h euglycemic hyperinsulinemic clamp. RESULTS: At baseline, serum MIC-1 was higher in AN in comparison with other groups (normal-weight, P = 0.018; obese, P = 0.01). Hyperinsulinemia resulted in a significant increase in serum MIC-1 concentration in the entire study population (P < 0.001) and in AN (P < 0.001), normal-weight (P = 0.002) and obese (P < 0.001) groups analysed separately. Postclamp serum MIC 1 was still higher in AN women in comparison with other groups (normal-weight, P = 0.012; obese, P = 0.023). When normal-weight and obese women were analysed together, with the exclusion of AN group, an inverse correlation between insulin sensitivity and the change in serum MIC-1 during the clamp was observed (r = 0.27, P = 0.042). CONCLUSIONS: Hyperinsulinemia resulted in a significant increase in serum MIC-1 in different states of adiposity. Increased serum MIC-1 in AN women might be an additional factor responsible for weight loss in this group. PMID- 21645024 TI - Coronary artery disease is associated with higher epicardial retinol-binding protein 4 (RBP4) and lower glucose transporter (GLUT) 4 levels in epicardial and subcutaneous adipose tissue. AB - OBJECTIVE: Retinol-binding protein 4 (RBP4), produced by adipocytes and hepatocytes, contributes to an unfavourable lipid profile and insulin resistance, which can contribute to the development of coronary artery disease (CAD). Recently, several studies have shown that epicardial adipose tissue (EAT) differs from subcutaneous adipose tissue (SAT) and plays a role on the physiopathology of CAD because of its proximity to the coronary arteries. We aimed to study the expression and secretion levels of RBP4 in both fat tissues and explore its possible association with CAD. RESEARCH DESIGN AND METHODS: Fifty-eight patients undergoing heart surgery were included in the study. We analysed RBP4 mRNA expression by real-time PCR, protein expression by Western blot and immunohistochemistry, and secretion of EAT and SAT explants from CAD and non-CAD patients by Enzyme Immunoassay. RESULTS: Retinol-binding protein 4 is expressed at similar levels in EAT and SAT, mainly from adipocytes. Protein levels were higher in EAT from CAD than non-CAD patients (0.63 +/- 0.09 arbitrary units (a.u).; n = 10) vs (0.41 +/- 0.04 a.u.; n = 13, P = 0.039). In contrast, GLUT4 mRNA levels were lower in EAT from CAD than non-CAD patients (6.55 +/- 0.16 a.u.; n = 13) vs (7.21 +/- 0.18 a.u.; n = 14, P = 0.012). We also found differential expression in SAT between samples from CAD and non-CAD patients [(6.63 +/- 0.16 a.u.; n = 14) vs (7.21 +/- 0.14 a.u.; n = 14, P = 0.009)]. Besides, EAT releases higher RBP4 levels than SAT after 3, 6, 24 and 48 h of culture. These levels were independent of CAD but significantly higher in diabetic than nondiabetic patients. CONCLUSION: Retinol-binding protein 4 levels behave differently in EAT and SAT with respect to CAD. However, both adipose tissues have lower GLUT4 levels in patients with CAD. These findings suggest a differential regulation of RBP4 production in EAT and SAT that may be influenced by local factors. PMID- 21645025 TI - Activating mutations in the calcium-sensing receptor: genetic and clinical spectrum in 25 patients with autosomal dominant hypocalcaemia - a German survey. AB - OBJECTIVE: Autosomal dominant hypocalcaemia or hypoparathyroidism is caused by activating mutations of the calcium-sensing receptor (CaSR). Treatment with calcium and vitamin D often worsens hypercalciuria and nephrocalcinosis, and renal impairment can result. Our aim was to describe the phenotypic variance of this rare disorder in a large series and to evaluate the outcome after long-term treatment. DESIGN: Nationwide retrospective collaborative study. PATIENTS: We describe 25 patients (14 men and 11 women), 20 belonging to 11 families and five single cases. MEASUREMENTS: Activating CaSR mutations and clinical and biochemical findings were evaluated. RESULTS: Nine different missense mutations of the CaSR, including one novel variant (M734T), were found. Twelve patients (50%) were symptomatic, 9 (36%) had basal ganglia calcifications and 3 (12%) had nephrocalcinosis. Serum calcium was decreased (1.87 +/- 0.13 mm), and PTH was decreased (n = 19) or inappropriately low (n = 4). The occurrence of hypocalcaemic symptoms at diagnosis was related to the degree of hypocalcaemia. The occurrence of features like calcification of basal ganglia or kidney calcification did not correlate with the severity of hypocalcaemia or the age at diagnosis. The most common treatment was calcitriol (median dosage 0.6 MUg/day), and the mean duration of therapy was 7.1 years (max. 26 years). Hypercalcaemic episodes rarely occurred, and the rate of kidney calcifications was remarkably low (12%). CONCLUSION: This series increases the limited knowledge of mutations and phenotypes of this rare disorder. Mutation analysis of the CaSR gene facilitates patient and family management. Low dosages of calcitriol resulted in less frequent renal calcifications. PMID- 21645026 TI - Expression of interleukin-22 in Sjogren's syndrome: significant correlation with disease parameters. AB - Sjogren's syndrome (SS) is an autoimmune disease targeting the exocrine glands resulting in xerostomia/keratoconjunctivitis sicca. Presently, we examined the levels and clinical correlations of IL-22 in SS. Patients with SS together with normal controls were randomly selected. IL-22 was detected at significantly higher levels in sera of patients with SS. The levels of IL-22 present in sera showed statistically significant direct correlations with hyposalivation, anti SSB, anti-SSA/SSB combined, hypergammaglobulinemia and rheumatoid factor. IL-22 showed a direct correlation with major clinical parameters. The data suggest that IL-22 plays a critical role in the development of SS, and further study is needed to examine its function in human SS. PMID- 21645027 TI - Periodontopathogen- and host-derived immune response in acute coronary syndrome. AB - Owing to molecular mimicry, periodontal pathogen carriage may result in a systemic cross-reactive immune response with the host. The analyses were performed to investigate if serum antibody levels to human heat shock protein 60 (HSP60) are associated with the antibody levels and salivary carriage of two periodontal pathogens, Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis, as well as with the dental status in patients with acute coronary syndrome (ACS). ACS patients (n = 141) were monitored at baseline when entering to hospital, and after 1 week, 3 months and 1 year. Periodontal status was recorded by dental radiographs, and A. actinomycetemcomitans and P. gingivalis were detected by PCR from saliva at baseline. Serum IgG and IgA antibody levels were determined at all time points. All antibody levels remained quite stable during the follow-up. Serum IgG-class antibody levels to A. actinomycetemcomitans and HSP60 had a strong positive correlation with each other at all time points (r~0.4, P < 0.05). Mean serum IgG antibody levels to HSP60 were significantly higher in the A. actinomycetemcomitans IgG- and IgA-seropositive than in the seronegative patients, but did not differ between the pathogen carriers compared to the non-carriers. HSP60 antibody levels did not differ significantly between the edentulous, non-periodontitis and periodontitis patients. Despite the observed cross-reactivity in the systemic IgG-class antibody response to HSP60 and A. actinomycetemcomitans, the pathogen carriage in saliva or the periodontal status did not affect the HSP60 antibody levels in ACS patients. PMID- 21645029 TI - Reverse transcription real-time PCR for detection of porcine interferon alpha and beta genes. AB - A few studies provided convincing evidence of constitutive expression of type I interferons (IFNs) in humans and mice, and of the steady-state role of these cytokines under health conditions. These results were later confirmed in pigs, too. In line with this tenet, low levels of IFN-alpha/beta can be detected in swine tissues in the absence of any specific inducer. These studies are compounded by the utmost complexity of type I IFNs (including among others 17 IFN alpha genes in pigs), which demands proper research tools. This prompted us to analyse the available protocols and to develop a relevant, robust, reverse transcription (RT) real-time polymerase chain reaction (PCR) detection system for the amplification of porcine IFN-alpha/beta genes. The adopted test procedure is user-friendly and provides the complete panel of gene expression of one subject in a microtitre plate. Also, a proper use of PCR fluorochromes (SYBR((r)) versus EvaGreen((r)) supermix) enables users to adopt proper test protocols in case of low-expression porcine IFN-alpha genes. This is accounted for by the much higher sensitivity of the test protocol with EvaGreen((r)) supermix. Interestingly, IFN beta showed the highest frequency of constitutive expression, in agreement with its definition of 'immediate early' gene in both humans and mice. Results indicate that the outlined procedure can detect both constitutively expressed and virus-induced IFN-alpha/beta genes, as well as the impact of environmental, non infectious stressors on the previous profile of constitutive expression. PMID- 21645028 TI - Pattern of pre-existing IgG subclass responses to a panel of asexual stage malaria antigens reported during the lengthy dry season in Daraweesh, Sudan. AB - The anti-malarial IgG immune response during the lengthy and dry season in areas of low malaria transmission as in Eastern Sudan is largely unknown. In this study, ELISA was used for the measurement of pre-existing total IgG and IgG subclasses to a panel of malaria antigens, MSP2-3D7, MSP2-FC27, AMA-1 and Pf332 C231. The results showed that the antibody responses were predominantly age dependent, antigen specific, and their lifespan was at least 5-6 month long. Generally, the IgG3 was most abundant IgG subclass, and the most recognized antigen was Pf332-C231. Furthermore, the correlation between the levels of IgG subclasses was strongest between IgG1 and IgG3, which were more predictive to the total IgG levels. Finally, the response pattern of each of the IgG subclasses to the different test antigens that were spanning the dry season and the correlation between these responses were described in details for the first time. PMID- 21645030 TI - Anaphylactic reactions in mice with Fenugreek allergy. AB - Fenugreek is a legume mostly used as a spice in Indian-style cooking. Although it has been used since ancient times, its allergenicity has only been reported in the last two decades. It poses special problems as an emerging and often hidden allergen. Fenugreek exposure may have serious implications also for individuals with peanut allergy because of cross-reactivity. Because a new food requires a model specially designed for that particular food, the aim of our study was to develop a food allergy model of fenugreek in mice with anaphylaxis as the endpoint. Mice were immunized perorally using cholera toxin as adjuvant. A two compartment response surface design with immunoglobulin (Ig)E as the main variable was used to estimate the optimal sensitizing dose of fenugreek, which was further used to evaluate the model. The mice were challenged perorally with a high dose of fenugreek, and signs of anaphylactic reactions were observed. Challenged mice showed high levels of mouse mast cell protease-1, developed specific IgE against several proteins in the fenugreek extract, had elevated levels of IgG1 and IgG2a and showed a general shift towards a Th2 response as determined by ex vivo production of cytokines. Total IgE levels were substantially decreased after challenge. In conclusion, we have established a mouse model of IgE-mediated fenugreek allergy demonstrating anaphylactic reactions upon challenge. There is little information on fenugreek cross-allergy to other legumes like peanut, soy and lupin, and we expect that this model will be a valuable tool in further research on legume allergy. PMID- 21645031 TI - Novel IFNgamma ELISPOT assay for detection of functional carcinoembryonic antigen specific chimeric antigen receptor-redirected T cells. AB - We here describe the development of a novel ELISPOT assay for the detection and enumeration of IFNgamma-secreting functional chimeric antigen receptor (CAR) redirected T cells against carcinoembryonic antigen (CEA). This method is valuable for clinical trials to monitor the presence of functional CEA-specific T cells transduced with a CAR. The same principle should be applicable for the detection of functional CAR-redirected T cells against any other tumour associated antigens by immobilizing a particular biotinylated antigen to streptavidin-coated beads. PMID- 21645032 TI - Lysosomal membrane permeabilization induces cell death in human mast cells. AB - Mast cells (MC) have pathogenic roles in numerous disorders, and strategies that stabilize MC or induce MC apoptosis are therefore emerging as possible therapeutic regimens. A typical feature of MC is their high content of secretory lysosomes (granules), containing numerous components such as biogenic amines, cytokines, serglycin proteoglycan and proteases. Damage to the secretory lysosomes will thus lead to leakage of these compounds, including the proteases, into the cytosol, and this could potentially trigger apoptosis. Here, we evaluated whether MC are sensitive to cell death induced by secretory lysosome destabilization, induced by the lysosomotropic agent Leu-Leu-OMe (LLME). Human MC were sensitive to LLME-induced cell death. In contrast, fibroblasts and HEK-293 cells were largely resistant. As judged by Annexin V/propidium iodide staining, LLME caused apoptotic cell death, and this was supported by induction of caspase 3-like activity, detection of activated caspase-3 by immunoblot analysis and reduced cell death in the presence of a caspase inhibitor. In support of a role for serglycin in regulating LLME-induced cell death, the survival rate of various cell types correlated negatively with the level of serglycin expression. In summary, this study introduces the concept of using lysosomotropic agents to induce cell death of human MC. PMID- 21645033 TI - Spindle cell lipoma with collagen rosettes. PMID- 21645034 TI - Amyloidosis cutis dyschromica associated with atypical Parkinsonism, spasticity and motor weakness in a Pakistani female. AB - Amyloidosis cutis dyschromica is a rare form of cutaneous amyloidosis in which there is deposition of keratinocyte-derived amyloid with involvement of almost the entire integument, leading to diffuse dyschromia without associated systemic abnormalities. We report the case of a 40-year-old female who presented with the onset of diffuse hyperpigmentation shortly after birth, which was followed by the widespread development of numerous 2-5 mm hypopigmented macules. Biopsy of the one of these macules revealed eosinophilic globular material in the papillary dermis with Congo red birefringence which also stained positively for high molecular weight cytokeratin. Electron microscopy confirmed the presence of 11 nm hollow fibrils, consistent with amyloid. Similar clinical changes were noted in a younger male sibling. Both patients also suffered from an unexplained neurological disorder characterized by atypical Parkinsonism, spasticity and motor weakness. This association has not been shown before and may represent a heretofore unreported contiguous gene syndrome. PMID- 21645035 TI - Lymph nodal Merkel cell carcinoma: primary tumor or metastasis from unknown primary site? PMID- 21645036 TI - The frequency of dual TCR-PCR clonality in granulomatous disorders. AB - BACKGROUND: A granulomatous infiltrate in association with cutaneous T-cell lymphoma is uncommon. The diagnosis of mycosis fungoides can be difficult in the setting of an exuberant granulomatous infiltrate that obscures the neoplastic lymphoid infiltrate, thereby mimicking a granulomatous dermatitis. Therefore, the clinical context and supplemental molecular analysis, such as the demonstration of a monoclonal T-cell population, may assist in diagnosis. Monoclonal T-cell populations have been reported in association with inflammatory conditions and serve as a diagnostic pitfall. The frequency of T-cell clonality in association with granulomatous dermatitides has not yet been established. METHODS: We identified 29 patients with granulomatous dermatitis who had biopsies at two distinct body sites. Results were correlated with clinical follow up and with clonal T-cell receptor-gamma chain rearrangement as detected by polymerase chain reaction-based analysis (dual TCR-PCR). RESULTS: Clinical follow up was obtained in 17 of 29 cases (58.6%). Twenty-five of 29 cases of granulomatous dermatitis lacked T-cell monoclonality. Three cases of granuloma annulare contained a T-cell clone in one of the two biopsies. One case of necrobiotic xanthogranuloma showed an identical T-cell clone in multiple biopsies. CONCLUSIONS: The use of dual TCR PCR analysis, that is, T-cell clonality analysis in biopsy specimens from two different sites, serves as an adjunct to assist in distinguishing granulomatous inflammatory reactions from granulomatous T-cell lymphoma, including granulomatous mycosis fungoides. The occasional finding of a T-cell clone in a granulomatous dermatitis underscores the importance of clinicopathological correlation in daily diagnosis. PMID- 21645038 TI - CD117 (c-KIT) staining in desmoplastic melanoma. PMID- 21645037 TI - Polypoid Spitz nevus: two cases evaluated with genetic technique, prolonged follow up, and sentinel lymph node biopsy. AB - Polypoid Spitz nevus represents a spitzoid melanocytic neoplasm that frequently has worrying and challenging histopathological details. Distinction from polypoid melanoma may not be straightforward. Two cases of polypoid Spitz nevus with striking histopathological features were studied. One case had prolonged follow up (Case 1) and one patient had undergone sentinel lymph node biopsy (Case 2), and fluorescence in situ hybridization (FISH) analysis was also completed (both cases). Follow up and genetic analysis of three control cases of polypoid melanoma is also presented. Our clinical and genetic results suggest that both the polypoid Spitz nevi were benign. The patients are alive with no evidence of disease. FISH analysis did not show abnormalities with probes tested. This is in sharp contrast with the control cases of polypoid melanoma, wherein genomic alterations were detectable. Our data indicate that the two polypoid lesions presented here are most probably benign, despite their worrying histopathological features. More cases with long-term follow up and greater numbers of DNA probes are necessary to extend this conclusion to other ambiguous melanocytic tumors. PMID- 21645040 TI - Self-efficacy and its influence on recovery of patients with stroke: a systematic review. AB - AIMS: To provide an overview of the literature focusing on the influence of self efficacy and self-efficacy enhancing interventions on mobility, activities of daily living, depression and quality of life of patients with stroke. BACKGROUND: There is growing evidence for the importance of self-efficacy in the care of people with enduring illness. Therefore, it is important to describe the association of self-efficacy and patient outcomes and the evidence for the effects of self-efficacy interventions for stroke patients. DATA SOURCES: Studies were retrieved from a systematic search of published studies over the period of 1996-2009, indexed in the Cumulative Index to Nursing and Allied Health Literature, Medline, Psychinfo and Embase and focusing on stroke, the influence of self-efficacy and self-efficacy enhancing interventions. METHODS: A systematic review was carried out. Studies were critically appraised and important characteristics and outcomes were extracted and summarized. RESULTS: Seventeen articles were included in the review. Self-efficacy was positively associated with mobility, activities of daily living and quality of life and negatively associated with depression. Four self-efficacy interventions were identified. The evidence for the effects of these interventions was inconclusive. CONCLUSIONS: Patients with high self-efficacy are functioning better in daily activities than patients with low self-efficacy. The evidence concerning the determinants influencing self-efficacy and the self-efficacy interventions makes clear how nurses can develop and tailor self-efficacy interventions for the clinical practice of people with stroke. Therefore, it is necessary to further emphasize the role of self-efficacy in the care for stroke patients in the nursing curriculum. PMID- 21645039 TI - Neuroprotective effects of oestrogen against oxidative toxicity through activation of G-protein-coupled receptor 30 receptor. AB - 1. 17-beta-oestradiol (E2) plays a critical role in neuroprotection through both genomic and non-genomic mechanisms. The aim of the present study was to investigate the role of G-protein-coupled receptor 30 (GPR30), a new kind of oestrogen receptor, in the neuroprotection against oxidative insult. 2. The neuroprotection evoked by GPR30 stimulation was examined in cultured cortical neurons. Hoechst 33258/propidium iodide double staining, flow cytometric analysis and western blotting were applied to assess neuronal apoptosis induced by H(2)O(2) . 3. We found that the GPR30 agonist, G1, and E2 attenuated apoptosis induced by H(2)O(2) exposure. Furthermore, G1 (1 nmol/L) or E2 (1 nmol/L) significantly increased the levels of phosphorylated extracellular signal regulated kinase 1/2 (p-ERK1/2), Bcl-2 and pro-caspase-3. Pretreatment with ICI182780, a highly selective nuclear oestrogen receptor antagonist that is used to block the classical ERalpha and ERbeta receptors, did not totally block the neuroprotective effects of E2 (1 nmol/L) and had no effect on the neuroprotective effects of G1 (1 nmol/L). 4. Our data suggest that GPR30 is involved in the neuroprotection against oxidative insult. The neuroprotection evoked by GPR30 stimulation was associated with the signalling through the ERK1/2 kinase pathway. In addition, the anti-apoptotic activity of GPR30 was dependent on the expression of Bcl-2 and pro-caspase-3. GPR30 might be a potential therapeutic target for neuroprotection and oxidative stress. PMID- 21645041 TI - The effect of music on biochemical markers and self-perceived stress among first line nurses: a randomized controlled crossover trial. AB - AIM: The aim of this study was to examine the effects of music on stress indices and to examine the association between music preference and stress. BACKGROUND: Although clinical studies have demonstrated the effectiveness of music on stress, study results have been inconsistent. At the time of writing, no known publications had investigated the effects of preferred music on workers in high stress professions such as nursing. METHODS: Using a randomized crossover controlled trial, 54 nurses were randomly assigned to a music/chair rest sequence or chair rest/music sequence during the period February to June 2006. Each intervention lasted for 30 minutes. Participants in the music condition listened to self-selected soothing music using headphones for 30 minutes. In the chair rest condition, participants sat quietly for 30 minutes. Serial measurements of participants' heart rate, mean arterial pressure, finger temperature and cortisol levels were taken with a BP monitor and chemillumincent immunoassay every 15 minutes throughout the procedure. FINDINGS: Compared with chair rest, participants had a lower perceived stress level, cortisol, heart rate, mean arterial pressure and higher finger temperature while listening to music (P < 0.05). Significant differences were also found between the two conditions in terms of post-test heart rate, cortisol levels, finger temperature and mean arterial pressure (P < 0.05). Music preference scores ranged between 7 and 10, with a mean score of 8.81 (sd = 1.05), and was significantly associated with mean arterial pressure, cortisol levels, self-perceived stress and finger temperature. CONCLUSION: The findings provided evidence for nurses to use soothing music as a research-based nursing intervention for stress reduction. PMID- 21645042 TI - Associations between self-rated health and psychosocial conditions, lifestyle factors and health resources among hospital nurses in Lithuania. AB - AIM: The aim of our study was to investigate the association between self-rated health and psychosocial factors at work and everyday life (job demands, job control, social support, workplace bullying, life-threatening events); health behaviours (smoking, alcohol, being overweight, obesity, low physical activity); mental distress; job satisfaction; and sense of coherence in a representative sample of Lithuanian hospitals' internal medicine department nurses. BACKGROUND: Recent trends to extend the retirement age in many countries of the European Union challenge future public health. Nurses are exposed to a broad variety of adverse psychosocial factors at work and in every day life that affect their health perception. As the retirement age in Lithuania is to be extended to 65 years for women, research on the associations of poor self-rated health with related factors is important. METHOD: A cross-sectional study was carried out in the period 2005-2006 using the representative sample of Lithuanian hospitals' internal medicine department nurses. Data were collected from 748 nurses using questionnaire (response rate 53.9%). RESULTS: About 60.4% of nurses rated their health negatively. In the fully adjusted model age, high job demands, low job control, low social support at work, life-threatening events, low physical activity, being overweight, obesity, mental distress, job dissatisfaction and weak sense of coherence were associated with negative self-rated health. CONCLUSIONS: Preventive strategies against adverse psychosocial working conditions of nurses should be implemented in the Lithuanian hospitals. PMID- 21645043 TI - Differences in intensive care unit work environments among and within hospitals using subscales and a composite measure of the Revised Nursing Work Index. AB - AIM: To examine variations in nurses' perceptions of their work environments among hospitals and intensive care units, and to compare analytic findings from using subscales and a composite measure of the Revised Nursing Work Index at the hospital and intensive care unit levels. BACKGROUND: A positive relationship has been found between the nurse work environment and outcomes for patients and nurses. Nurses' perceptions of their work environments have been analysed using different analytic approaches. METHODS: A survey was conducted in August-October 2007 that included 817 staff nurses in 39 adult intensive care units of 15 hospitals in South Korea. Seven subscales of the Revised Nursing Work Index were identified from an exploratory factor analysis. The subscales and composite (mean of the seven subscales) for each hospital and intensive care unit were analysed using multilevel regression analyses and classified as good, moderate or poor environments. RESULTS: Considerable variations in the subscales were found among both hospitals and intensive care units. On the composite measure, 2 hospitals were rated as good, 10 moderate and 3 poor; 9 intensive care units were ranked as good, 24 moderate and 6 poor. Even intensive care units within hospitals exhibited variations in the subscales and composite. Most hospitals and intensive care units had mixed (i.e., good, moderate, poor) environments across the seven subscales and thus, subscales were not always congruent with the composite. CONCLUSION: Heterogeneity of the subscales and the composite measure, and the differences among intensive care units within hospitals imply that use of different analytic approaches may reveal different findings and perspectives of nurse work environments. PMID- 21645044 TI - Older adults' perceptions of feeling safe in an intensive care unit. AB - AIM: This paper is a report of a study of older adults' perceptions of feeling safe in an intensive care unit. BACKGROUND: Significant ageing of the world population is expected. Correspondingly, there is a growing global urgency for healthcare systems to prepare for the increased number of older adults who will require intensive care. Although patients have recounted situations in which feeling safe was essential to their experience, there is little empirical evidence about feeling safe for older adults in intensive care. METHOD: In 2008, 10 older adults who suffered unexpected critical health events and who were admitted to intensive care units were recruited for this grounded theory study. Participants were interviewed after transferring to an intermediate care unit and again after discharge. They were asked to reflect upon and discuss their experiences of feeling safe in intensive care. FINDINGS: Participants' actual and expected interaction with intensive care nurses was essential to their perceptions of feeling safe. Four main categories were identified as requisite to interaction with nurses: proximity, oversight, predictability and initiative. A substantive grounded theory of feeling safe for older adults in intensive care is presented. CONCLUSION: Nurses must present themselves as accessible so patients perceive that they can initiate interaction with a qualified nurse. Patients must also see that nurses are monitoring them and believe that the nurses will help them if they experience a critical health event while in the intensive care unit. PMID- 21645045 TI - The impact of psychological capital on job embeddedness and job performance among nurses: a structural equation approach. AB - AIMS: The objective of this study was to provide empirical evidence on the relationships between psychological capital, job embeddedness and performance. This paper also seeks to present the theoretical development of psychological capital and job embeddedness in nursing research and their application to nursing practices. BACKGROUND: Psychological capital was recently identified as a core construct in the literature of positive psychology. However, there is considerably less evidence on its positive effects on job embeddedness and performance among nursing personnel. METHODS: Questionnaires were distributed to approximately 1000 nurses employed in five university hospitals in Heilongjiang province in China. Data were collected in november 2009. the response rate was 73.3%. structural equation modelling was employed to test the proposed relationships. FINDINGS: The results support the hypothesized model. This research outlined a strong relationship between the self-reported psychological capital, job embeddedness and performance of the nurses. The study findings suggest that improving the individual-accumulated psychological state of nurses will have a positive impact on their retention intention and job performance. CONCLUSIONS: These findings suggest that higher psychological capital increases the self-reported job embeddedness and performance of these nurses. PMID- 21645046 TI - Barriers and facilitators to optimize function and prevent disability worsening: a content analysis of a nurse home visit intervention. AB - AIMS: This paper is a report of an analysis of how to better understand the results of the nurse home visit intervention in the Medicare Primary and Consumer Directed Care Demonstration in terms of facilitators and barriers to disability improvement/maintenance as compared with disability worsening. BACKGROUND: There is a lack of literature describing how nurse home visit interventions are able to maintain/improve disability among older persons with disability. The present study is one of only six reporting beneficial disability outcomes. METHODS: Cases were purposefully sampled to represent change in the disability construct leading to selection of ten cases each of disability maintenance/improvement (no change or decrease in total Activities of Daily Living score from baseline) and worsening (an increase in total Activities of Daily Living score from baseline). Data from nurses' progress notes and case studies (collected in March 1998-June 2002) were analysed using qualitative descriptive analysis (May 2009). These results remain relevant because the present study is one of the few studies to identify select nurse activities instrumental in postponing/minimizing disability worsening. RESULTS/FINDINGS: Three primary themes captured the facilitators and barriers to effective disability maintenance/improvement: (1) building and maintaining patient-centred working relationships, (2) negotiating delivery of intervention components and (3) establishing balance between patients' acute and chronic care needs. Sub-themes illustrate nurse, patient and system factors associated with effective disability maintenance/improvement (e.g. nurse caring, communicating, facilitating interdisciplinary communication) and barriers associated with disability worsening (e.g. dementia, depression and recurring acute illnesses). CONCLUSION: This study provides new insights about the facilitators and barriers to effective disability maintenance/improvement experienced by patients receiving home visits. Potential opportunities exist to integrate these insights into best-practice models of nurse home visiting. PMID- 21645047 TI - Qualitative studies using in-depth interviews with older people from multiple language groups: methodological systematic review. AB - AIM: This paper is a report of a methodological review of language appropriate practice in qualitative research, when language groups were not determined prior to participant recruitment. BACKGROUND: When older people from multiple language groups participate in research using in-depth interviews, additional challenges are posed for the trustworthiness of findings. This raises the question of how such challenges are addressed. DATA SOURCES: The Cumulative Index to Nursing and Allied Health Literature, Scopus, Embase, Web of Science, Ageline, PsycINFO, Sociological abstracts, Google Scholar and Allied and Complementary Medicine databases were systematically searched for the period 1840 to September 2009. The combined search terms of 'ethnic', 'cultural', 'aged', 'health' and 'qualitative' were used. REVIEW METHODS: In this methodological review, studies were independently appraised by two authors using a quality appraisal tool developed for the review, based on a protocol from the McMaster University Occupational Therapy Evidence-Based Practice Research Group. RESULTS: Nine studies were included. Consideration of language diversity within research process was poor for all studies. The role of language assistants was largely absent from study methods. Only one study reported using participants' preferred languages for informed consent. CONCLUSION: More examples are needed of how to conduct rigorous in-depth interviews with older people from multiple language groups, when languages are not determined before recruitment. This will require both researchers and funding bodies to recognize the importance to contemporary healthcare of including linguistically diverse people in participant samples. PMID- 21645048 TI - Intensive care nurses' perceptions of their professional competence in the organ donor process: a national survey. AB - AIMS: This paper is a report of a study that explored Norwegian intensive care nurses' perceptions of their professional competence to identify educational needs in the organ donor process. BACKGROUND: Intensive care professionals are requested to consider organ donation each time they care for patients with severe cerebral lesion to ensure donor organs for transplantation. The donor process challenges intensive care nurses' professional competence. Nurses' knowledge and experience may influence their professional competence in caring for organ donors and their relatives. METHODS.: A cross-sectional survey was conducted in all 28 Norwegian donor hospitals between October 2008 and January 2009. Intensive care nurses (N = 801) were invited to participate and the response rate was 71.4%. Dimensions of professional competence, learning needs and contextual and demographic variables were explored. Data were analysed using descriptive and inferential statistics. FINDINGS: Few intensive care nurses had extensive experience of or competence and training in organ donation. Nurses working at university hospitals had more experience, but lesser training than nurses in local hospitals. Experience of donor acquisition had an impact on intensive care nurses' perceptions of their professional competence in the donor process. Discussions on the ward and educational input were seen as important for the further development of professional competence. CONCLUSION: Training provided by experienced colleagues and a culture that encourages discussion about aspects of the donor process can develop nurses' professional competence and communally defined professional practice. Educational input that cultivates various types of knowledge can be beneficial in organ donation. PMID- 21645049 TI - The N-terminal third of the BinB subunit from the Bacillus sphaericus binary toxin is sufficient for its interaction with midgut receptors in Culex quinquefasciatus. AB - Heterodimeric binary (Bin) toxin, the major insecticidal protein from Bacillus sphaericus, acts on Culex quinquefasciatus larvae through specific binding to the midgut receptor Cqm1, a role mediated by its 448-amino-acid-long BinB subunit. The molecular basis for receptor recognition is not well understood and this study attempted to identify protein segments and amino acid motifs within BinB that are required for this event. First, N- and C-terminally truncated constructs were evaluated for their capacity to bind to native Cqm1 through pull-down assays. These showed that residues N33 to L158 of the subunit are required for Cqm1 binding. Nine different full-length mutants were then generated in which selected blocks of three amino acids were replaced by alanines. In new pull-down assays, two mutants, in which residues (85) IRF(87) and (147) FQF(149) were targeted, failed to bind the receptor. Competition binding assays confirmed the requirements for the N-terminal 158 residues, and the (147) FQF(149) epitope, for the mutant proteins to compete with native Bin toxin when binding to membrane fractions from the insect midgut. The data from this work rule out the involvement of C-terminal segments in receptor binding, highlighting the need for multiple elements within the protein's N-terminal third for it to occur. PMID- 21645050 TI - The gyrB gene is a useful phylogenetic marker for exploring the diversity of Flavobacterium strains isolated from terrestrial and aquatic habitats in Antarctica. AB - Within the phylum Bacteroidetes, the gyrB gene, encoding for the B subunit of the DNA gyrase, has been used as a phylogenetic marker for several genera closely related to Flavobacterium. The phylogenies of the complete 16S rRNA gene and the gyrB gene were compared for 33 Antarctic Flavobacterium isolates and 23 type strains from closely related Flavobacterium species. gyrB gene sequences provided a higher discriminatory power to distinguish between different Flavobacterium groups than 16S rRNA gene sequences. The gyrB gene is therefore a promising molecular marker for elucidating the phylogenetic relationships among Flavobacterium species and should be evaluated for all the other type strains of described Flavobacterium species. Combining the phylogeny of both genes, the new Antarctic Flavobacterium strains constitute 15 Flavobacterium groups, including at least 13 potentially new species together with one group of isolates probably belonging to the species Flavobacterium micromati and one group close to Flavobacterium gelidilacus. PMID- 21645051 TI - A unique transcription mode of rabies virus high egg passage-Flury strain detected in infected baby hamster kidney-21 cells. AB - The transcription mode of rabies virus high egg passage-Flury (HEP) strain was examined and compared with that of Evelyn Rokitniki Abelseth (ERA) strain by northern blot analysis using rabies virus gene-specific probes. The ERA strain was shown to exclusively produce monocistronic mRNAs in transcription. All combinations of multicistronic transcripts, including five monocistronic mRNAs, were detected in the viral RNA transcripts of HEP strain. It was concluded that the unique transcription mode is not due to the nucleotide structure of the genome RNA template, but rather to the viral RNA polymerase of HEP strain. The viral polymerase of HEP strain read through the gene junction at a high frequency. The HEP strain has been passaged many times in chick embryo and cultured cells, and has adapted to propagate well in the baby hamster kidney-21 (BHK-21) cells. Through these passages in various hosts, the HEP strain has acquired a unique transcription mode that might have an advantage in amplification of the virus. PMID- 21645052 TI - Possible participation of the Rho/Rho-associated coiled-coil-forming kinase pathway in the cell death of Cryptococcus neoformans caused by Staphylococcus aureus adherence. AB - We here report the apoptotic death of a fungus, Cryptococcus neoformans (C. neoformans), in response to adherence of the pathogenic bacterium Staphylococcus aureus (S. aureus). In co-culture, cryptococcal actin was visibly aggregated. To investigate the mechanism of death, the participation of small GTP(guanosine triphosphate)-binding proteins belonging to the Rho subfamily, which regulate the actin cytoskeleton, was explored. C. neoformans was cultured with S. aureus in the presence of N-(4-pyridyl)-4-(1-aminoethyl)cyclohexanecarboxamide (Y-27632), an inhibitor of Rho-associated coiled-coil forming kinase (ROCK), a downstream effector of Rho. Death of C. neoformans was significantly reduced by the inhibitor. Concomitantly, Y-27632 prevented the aggregation of actin. Therefore, it was concluded that the Rho/ROCK pathway is involved in cell death induced by adherence stress. Increased expression of the voltage-dependent anion channel (VDAC), located in the mitochondrial outer membrane, has previously been observed in the apoptosis-like death of C. neoformans in the presence of hydrogen peroxide. Ruthenium red (RuR), which binds to VDAC and inhibits cytochrome c release, was used to determine the involvement of VDAC following adherence stress caused by S. aureus. RuR treatment increased the viability of C. neoformans co cultured with S. aureus in a dose dependent manner. These findings suggest that Rho-ROCK signaling could be involved, via a mitochondrial pathway, in the apoptosis-like death of C. neoformans induced by the adherence of S. aureus. PMID- 21645053 TI - Strain-specific effects of reducing agents on the cell-free conversion of recombinant prion protein into a protease-resistant form. AB - The pathogenic isoform (PrP(Sc) ) of the host-encoded normal cellular prion protein (PrP(C) ) is believed to be the infectious agent of transmissible spongiform encephalopathies. Spontaneous conversion of alpha-helix-rich recombinant PrP into the PrP(Sc) -like beta-sheet-rich form or aggregation of cytosolic PrP has been found to be accelerated under reducing conditions. However, the effect of reducing conditions on PrP(Sc) -mediated conversion of PrP(C) into PrP(Sc) has remained unknown. In this study, the effect of reducing conditions on the binding of bacterial recombinant mouse PrP (MoPrP) with PrP(Sc) and the conversion of MoPrP into proteinase K-resistant PrP (PrP(res) ) using a cell-free conversion assay was investigated. High concentrations of dithiothreitol did not inhibit either the binding or conversion reactions of PrP(Sc) from five prion strains. Indeed, dithiothreitol significantly accelerated mouse-adapted BSE-seeded conversion. These data suggest that conversion of PrP(Sc) derived from a subset of prion strains is accelerated under reducing conditions, as has previously been shown for spontaneous conversion. Furthermore, the five prion strains used could be classified into three groups according to their efficiency at binding and conversion of MoPrP and cysteine-less mutants under both reducing and nonreducing conditions. The resulting classification is similar to that derived from biological and biochemical strain-specific features. PMID- 21645054 TI - Full sequence analysis of the original Sapporo virus. AB - In this study, the full-length genome sequence of the prototype of sapovirus, namely Sapporo virus (SV82), was identified. Sapporo virus RNA was extracted from a fecal sample, amplified by RT-PCR and the PCR products sequenced directly and analyzed. Sequence analysis showed that Sapporo virus consists of 7433 nucleotides and has three open reading frames. The Sapporo strain shows 91.7% nucleotide sequence identity to the Manchester virus. Phylogenic analysis has also revealed the closeness of Sapporo virus to other sapovirus/genogroup I strains. Basic information on the evolutionary history of sapovirus analysis is provided here. PMID- 21645086 TI - Melatonin's stimulatory effect on adult hippocampal neurogenesis in mice persists after ovariectomy. AB - In this study, we examined whether melatonin treatment would increase new cell formation in the hippocampus in ovariectomized (OVX) mice. Chronic exogenous melatonin administration increased bromodeoxyuridine (BrdU) (OVX-sham 72 +/- 3.2 versus OVX-mel 122 +/- 12.0; P < 0.05) and doublecortin (DCX) (OVX-sham 88 +/- 3.1 versus OVX-mel 176 +/- 9.9; P < 0.05) immunoreactive cells in the hippocampus of ovariectomized mice. This neuronal development was correlated with synaptic plasticity, identified using the Golgi impregnation method to quantify dendritic spines in mouse dentate gyrus (DG). Finally, the antidepressant-like state of the animals was evaluated by the tail suspension test. The results indicate that melatonin acts on birth, survival, and differentiation of new neurons in the hippocampus, stimulates maturation of spines, and exerts an antidepressant-like action under estrogen-deprived conditions, in both a strain- and gender independent manner, suggesting that this indoleamine may be useful in improving brain functions. PMID- 21645087 TI - Warming preterm infants in the delivery room: polyethylene bags, exothermic mattresses or both? AB - AIMS: To compare the admission temperature of infants treated with polyethylene bags alone to infants treated with exothermic mattresses in addition to bags in the delivery room. METHODS: We prospectively studied infants born at <31 weeks' gestation who were placed in bags at birth. Some infants were also placed on mattresses. Admission axillary temperatures were measured in all infants on admission to the neonatal intensive care. We compared the temperatures of infants treated with bags alone to those treated with mattresses and bags. RESULTS: We studied 43 infants: 15 were treated with bags while 28 were treated with a bag and mattress. Mean admission temperature was similar between the groups. Hypothermia and hyperthermia occurred more frequently in infants treated with a bag and mattress, and more infants treated with a bag had admission temperatures 36.5-37.5 degrees C. CONCLUSION: The use of exothermic mattresses in addition to polyethylene bags, particularly in younger, smaller newborns, may result in more hypothermia and hyperthermia on admission. A randomised controlled trial is necessary to determine which strategy results in more infants having admission temperatures in the normal range. PMID- 21645088 TI - Sense of coherence and health behaviour in adolescence. AB - AIM: To explore associations of Sense of coherence (SOC) with health behaviour and social competence among 15-year-old adolescents. METHODS: Study population is a prospective cohort of a randomized cluster sample of families and their first born children from south-western Finland in 1986-1987. In this study, cross sectional data of the 15-year-olds were used. The present data were based on mailed, pretested questionnaires. The outcome variable, SOC, was based on the 13 item scale of Antonovsky's Orientation to Life Questionnaire (OLQ). The principal explanatory variables were health behaviour, including experienced oral health, and social competence. The statistical analysis was performed using linear regression modelling. RESULTS: Strong SOC of adolescents associated significantly with lighter use of alcohol, being a non-smoker, better care of oral health and better social competence compared with the others. CONCLUSION: Sense of coherence is a useful tool for identifying adolescents in need of extra support and motivation for their health behaviour. PMID- 21645089 TI - Saccade, search and orient--the neural control of saccadic eye movements. PMID- 21645090 TI - Full reaction time distributions reveal the complexity of neural decision-making. AB - Measurement of the stochastic distribution of reaction time or latency has become a popular technique that can potentially provide precise, quantitative information about the underlying neural decision mechanisms. However, this approach typically requires data from large numbers of individual trials, in order to enable reliable distinctions to be made between different models of decision. When data are not plentiful, an approximation to full distributional information can be provided by using a small number of quantiles instead of full distributions - often, just five are used. Although this can often be adequate when the proposed underlying model is a relatively simple one, we show here that, with more complex tasks, and correspondingly extended models, this kind of approximation can often be extremely misleading, and may hide important features of the underlying mechanisms that only full distributional analysis can reveal. PMID- 21645091 TI - Contribution of the retino-tectal pathway to visually guided saccades after lesion of the primary visual cortex in monkeys. AB - Previous reports on 'blindsight' have shown that some patients with lesions of the primary visual cortex (V1) could localize visual targets in their scotoma with hand and/or eye movements without visual awareness. A role of the retino tectal pathway on residual vision has been proposed but the direct evidence for this still remains sparse. To examine this possibility, we inactivated the superior colliculus (SC) of unilateral V1-lesioned monkeys using microinjections of muscimol, and analysed the effects on visually guided saccades. Following muscimol injections into the contralesional SC, the monkeys performed the visually guided saccade task with relatively minor deficits. The effects of ipsilesional SC inactivation were more severe. After injections, the monkeys failed to localize the target within the visual field represented at the injection site on the SC map. The effects of ipsilesional SC inactivation may result from sensory deficits, motor deficits or a combination of both. To examine these possibilities, we tested the effects of SC inactivation on the motor system by investigating spontaneous saccades. After inactivation of the ipsilesional SC, spontaneous saccades toward the injection site were not abolished, suggesting that impairment of visually guided saccades following inactivation of the ipsilesional SC could not be explained solely by a motor deficit and was primarily due to a visual deficit, presumably by interfering with processing in the superficial layer. We conclude that the retino-tectal pathway plays an essential role in residual vision after V1 lesion. The results suggest that this pathway may be involved in mediating unconscious vision in blindsight patients. PMID- 21645093 TI - The relationship between spatial attention and saccades in the frontoparietal network of the monkey. AB - Spatial attention bears a remarkable resemblance to saccadic eye movements from both a behavioural and a neurophysiological point of view. In this review, we examine the contributions of two cortical areas, namely the lateral intraparietal area (LIP) and frontal eye field (FEF), to saccades and attention and discuss the possible interactions between these two areas. Based on the results of electrophysiological studies and on inactivation studies performed in the macaque monkey, we propose that LIP is mainly involved in salience representation and an attentional selection mechanism that underlies saccade guidance, at least when two objects or locations are in competition in the visual environment. In contrast, we suggest that FEF is involved in coding and triggering saccadic eye movements, as well as in coding the location of attention or the attentional shifts. However, these two functions subserved by the FEF are dissociable at a neuronal level. Saccade planning and attentional selection are intimately coupled from a behavioural point of view but correspond to distinct functional operations. PMID- 21645092 TI - Control from below: the role of a midbrain network in spatial attention. AB - Spatial attention enables the brain to analyse and evaluate information selectively from a specific location in space, a capacity essential for any animal to behave adaptively in a complex world. We usually think of spatial attention as being controlled by a frontoparietal network in the forebrain. However, emerging evidence shows that a midbrain network also plays a critical role in controlling spatial attention. Moreover, the highly differentiated, retinotopic organization of the midbrain network, especially in birds, makes it amenable to detailed analysis with modern techniques that can elucidate circuit, cellular and synaptic mechanisms of attention. The following review discusses the role of the midbrain network in controlling attention, the neural circuits that support this role and current knowledge about the computations performed by these circuits. PMID- 21645094 TI - The role of the lateral intraparietal area in orienting attention and its implications for visual search. AB - Orienting visual attention is of fundamental importance when viewing a visual scene. One of the areas thought to play a role in the guidance of this process is the posterior parietal cortex. In this review, we will describe the way the lateral intraparietal area (LIP) of the posterior parietal cortex acts as a priority map to help guide the allocation of covert attention and eye movements (overt attention). We will explain the concept of a priority map and then show that LIP activity is biased by both bottom-up stimulus-driven factors and top down cognitive influences, and that this activity can be used to predict the locus of covert attention and initial saccadic latencies in simple visual search tasks. We will then describe evidence for how this system acts during covert visual search and how its activity could be used to optimize overt visual search performance. PMID- 21645096 TI - Investigating the role of the superior colliculus in active vision with the visual search paradigm. AB - We review here both the evidence that the functional visuomotor organization of the optic tectum is conserved in the primate superior colliculus (SC) and the evidence for the linking proposition that SC discriminating activity instantiates saccade target selection. We also present new data in response to questions that arose from recent SC visual search studies. First, we observed that SC discriminating activity predicts saccade initiation when monkeys perform an unconstrained search for a target defined by either a single visual feature or a conjunction of two features. Quantitative differences between the results in these two search tasks suggest, however, that SC discriminating activity does not only reflect saccade programming. This finding concurs with visual search studies conducted in posterior parietal cortex and the idea that, during natural active vision, visual attention is shifted concomitantly with saccade programming. Second, the analysis of a large neuronal sample recorded during feature search revealed that visual neurons in the superficial layers do possess discriminating activity. In addition, the hypotheses that there are distinct types of SC neurons in the deeper layers and that they are differently involved in saccade target selection were not substantiated. Third, we found that the discriminating quality of single-neuron activity substantially surpasses the ability of the monkeys to discriminate the target from distracters, raising the possibility that saccade target selection is a noisy process. We discuss these new findings in light of the visual search literature and the view that the SC is a visual salience map for orienting eye movements. PMID- 21645095 TI - Neural mechanisms of saccade target selection: gated accumulator model of the visual-motor cascade. AB - We review a new computational model developed to understand how evidence about stimulus salience in visual search is translated into a saccade command. The model uses the activity of visually responsive neurons in the frontal eye field as evidence for stimulus salience that is accumulated in a network of stochastic accumulators to produce accurate and timely saccades. We discovered that only when the input to the accumulation process was gated could the model account for the variability in search performance and predict the dynamics of movement neuron discharge rates. This union of cognitive modeling and neurophysiology indicates how the visual-motor transformation can occur, and provides a concrete mapping between neuron function and specific cognitive processes. PMID- 21645097 TI - Parietofrontal circuits in goal-oriented behaviour. AB - Parietal and frontal cortical areas play important roles in the control of goal oriented behaviour. This review examines how signal processing in the parietal and frontal eye fields is involved in coding and storing space, directing attention and processing the sensorimotor transformation for saccades. After a survey of the functional specialization of these areas in monkeys, we discuss homologous regions in the human brain in terms of topographic organization, storage capacity, target selection, spatial remapping, reference frame transformations and effector specificity. The overall picture suggests that bottom-up sensory, top-down cognitive signals and efferent motor signals are integrated in dynamic sensorimotor maps as part of a functionally flexible parietofrontal network. Neuronal synchronization in these maps may be instrumental in amplifying behaviourally relevant representations and setting up a functional pathway to route information in this parietofrontal circuit. PMID- 21645098 TI - The search for the neural mechanisms of the set size effect. AB - The set size effect in visual search refers to the linear increase in response time (RT) or decrease in accuracy as the number of distractors increases. Previous human and monkey studies have reported a correlation between set size and neural activity in the frontal eye field (FEF) and intraparietal sulcus (IPS). In a recent functional magnetic resonance imaging study, we did not observe a set size effect in the superior precentral sulcus (sPCS, thought to be the human homolog of the FEF) and IPS in an oculomotor visual search task (Ikkai et al., 2011). Our task used placeholders in the search array, along with the target and distractors, in order to equate the amount of retinal stimulation for each set size. We here attempted to reconcile these differences with the results from a follow-up experiment in which the same oculomotor visual search task was used, but without placeholders. A strong behavioral set size effect was observed in both studies, with very similar saccadic RTs and slopes between RT and set size. However, a set size effect was now observed in the sPCS and IPS. We comment on this finding and discuss the role of these neural areas in visual search. PMID- 21645099 TI - Split of spatial attention as predicted by a systems-level model of visual attention. AB - Can we attend to multiple distinct spatial locations at the same time? According to a recent psychophysical study [J. Dubois et al. (2009)Journal of Vision, 9, 3.1-11] such a split of spatial attention might be limited to short periods of time. Following N. P. Bichot et al. [(1999)Perception & Psychophysics, 61, 403 423] subjects had to report the identity of multiple letters that were briefly presented at different locations, while two of these locations (targets) were relevant for a concurrent shape comparison task. In addition to the design used by Bichot et al. stimulus onset asynchrony between shape onset and letters was systematically varied. In general, the performance of subjects was superior at target locations. Furthermore, for short stimulus onset asynchronies, performance was simultaneously increasing at both target locations. For longer stimulus onset asynchronies, however, performance deteriorated at one of the target locations while increasing at the other target location. It was hypothesized that this dynamic deployment of attention might be caused by competitive processes in saccade-related structures such as the frontal eye field. Here we simulated the task of Dubois et al. using a systems-level model of attention. Our results are consistent with recent findings in the frontal eye field obtained during covert visual search, and they support the view of a transient deployment of spatial attention to multiple stimuli in the early epoch of target selection. PMID- 21645100 TI - Contribution of the central thalamus to the generation of volitional saccades. AB - Lesions in the motor thalamus can cause deficits in somatic movements. However, the involvement of the thalamus in the generation of eye movements has only recently been elucidated. In this article, we review recent advances into the role of the thalamus in eye movements. Anatomically, the anterior group of the intralaminar nuclei and paralaminar portion of the ventrolateral, ventroanterior and mediodorsal nuclei of the thalamus send massive projections to the frontal eye field and supplementary eye field. In addition, these parts of the thalamus, collectively known as the 'oculomotor thalamus', receive inputs from the cerebellum, the basal ganglia and virtually all stages of the saccade-generating pathways in the brainstem. In their pioneering work in the 1980s, Schlag and Schlag-Rey found a variety of eye movement-related neurons in the oculomotor thalamus, and proposed that this region might constitute a 'central controller' playing a role in monitoring eye movements and generating self-paced saccades. This hypothesis has been evaluated by recent experiments in non-human primates and by clinical observations of subjects with thalamic lesions. In addition, several recent studies have also addressed the involvement of the oculomotor thalamus in the generation of anti-saccades and the selection of targets for saccades. These studies have revealed the impact of subcortical signals on the higher-order cortical processing underlying saccades, and suggest the possibility of future studies using the oculomotor system as a model to explore the neural mechanisms of global cortico-subcortical loops and the neural basis of a local network between the thalamus and cortex. PMID- 21645101 TI - Cortico-basal ganglia mechanisms for overcoming innate, habitual and motivational behaviors. AB - Most of the human behaviors are executed automatically under familiar circumstances. These behaviors are prepotent in that they take precedence over any other potential alternatives. Yet, humans are also capable of engaging cognitive resources to inhibit such a prepotent behavior and replace it with an alternative controlled behavior in response to an unforeseen situation. This remarkable capability to switch behaviors in a short period of time is the hallmark of executive functions. In this article, we first argue that the prepotent automaticity could emerge at least in three different domains - innate, habitual and motivational. We then review neurophysiological findings on how the brain might realize its switching functions in each domain, primarily by focusing on the monkey oculomotor system as the experimental model. Emerging evidence now suggests that multiple neuronal populations in the shared cortico-basal ganglia network contribute to overriding prepotent eye movement, be its origin innate, habitual or motivational. This consideration suggests the general versatility of the cortico-basal ganglia network as the neural mechanism whereby humans and other animals keep themselves from becoming subservient to reflex, habit and motivational impulses. PMID- 21645102 TI - Probing basal ganglia functions by saccade eye movements. AB - The basal ganglia (BG) are a group of subcortical structures involved in diverse functions, such as motor, cognition and emotion. However, the BG do not control these functions directly, but rather modulate functional processes occurring in structures outside the BG. The BG form multiple functional loops, each of which controls different functions with similar architectures. Accordingly, to understand the modulatory role of the BG, it is strategic to uncover the mechanisms of signal processing within specific functional loops that control simple neural circuits outside the BG, and then extend the knowledge to other BG loops. The saccade control system is one of the best-understood neural circuits in the brain. Furthermore, sophisticated saccade paradigms have been used extensively in clinical research in patients with BG disorders as well as in basic research in behaving monkeys. In this review, we describe recent advances of BG research from the viewpoint of saccade control. Specifically, we account for experimental results from neuroimaging and clinical studies in humans based on the updated knowledge of BG functions derived from neurophysiological experiments in behaving monkeys by taking advantage of homologies in saccade behavior. It has become clear that the traditional BG network model for saccade control is too limited to account for recent evidence emerging from the roles of subcortical nuclei not incorporated in the model. Here, we extend the traditional model and propose a new hypothetical framework to facilitate clinical and basic BG research and dialogue in the future. PMID- 21645103 TI - A perceptual discrimination task abnormally facilitates reflexive saccades in Parkinson's disease. AB - Numerous studies have shown that Parkinson's disease (PD) affects the ability to generate voluntary saccades and the ability to suppress reflexive saccades. The effects of PD on the generation of reflexive saccades, however, are not clear. Some studies report impairments, but there are also reports of abnormal facilitation or hyper-reflexivity of the saccade system in PD. Meanwhile, it has been reported that the concurrent performance of a perceptual discrimination task facilitates saccade initiation and reduces saccade latencies in healthy subjects [A. Montagnini & L. Chelazzi (2005)Vis. Res., 45, 3391-3401; L. Trottier & J. Pratt (2005)Vis. Res., 45, 1349-1354]. To investigate the circumstances under which the saccade system may appear hyper-reflexive in PD, we compared reflexive saccades with and without a concurrent perceptual discrimination task in 20 PD patients and 20 controls. Without the discrimination task, the PD group produced reflexive saccades at normal latencies. The discrimination task reduced saccade latencies more in the PD group than in the control group, resulting in abnormally short mean reflexive saccade latencies in the PD group. The discrimination task increased saccade gain in both groups, but saccades in the PD group remained hypometric as compared with saccades in the control group. We conclude that the attentional demands of this paradigm revealed a hypersensitivity to visual inputs in the PD group. PMID- 21645104 TI - Mechanisms for generating and compensating for the smallest possible saccades. AB - Microsaccades are small eye movements that occur during gaze fixation. Although taking place only when we attempt to stabilize gaze position, microsaccades can be understood by relating them to the larger voluntary saccades, which abruptly shift gaze position. Starting from this approach to microsaccade analysis, I show how it can lead to significant insight about the generation and functional role of these eye movements. Like larger saccades, microsaccades are now known to be generated by brainstem structures involved not only in compiling motor commands for eye movements, but also in identifying and selecting salient target locations in the visual environment. In addition, these small eye movements both influence and are influenced by sensory and cognitive processes in various areas of the brain, and in a manner that is similar to the interactions between larger saccades and sensory or cognitive processes. By approaching the study of microsaccades from the perspective of what has been learned about their larger counterparts, we are now in a position to make greater strides in our understanding of the function of the smallest possible saccadic eye movements. PMID- 21645105 TI - The role of the cerebellum in saccadic adaptation as a window into neural mechanisms of motor learning. AB - How does the nervous system guide the muscular periphery during the acquisition of a new motor skill? This is a fundamental question for researchers trying to understand the neural basis of motor learning. Recent advances in studying a valuable example of short-term motor learning, namely the adaptation of saccadic eye movements, have revealed neuronal processes in the cerebellum that underlie the unfolding of the learned behavior. In this review, we describe the latest findings from electrophysiology studies of saccadic adaptation and how they can generalize to more elaborate examples of cerebellum-dependent adaptation of movements. We focus our discussion on the plastic changes that are observed in the firing properties of Purkinje cells during the acquisition of the wanted motor response and describe how the altered activity of these neurons modifies the dynamics of the cerebellar microcircuitry. We finally demonstrate how such task-related modifications in the cerebellum are appropriate to fine-tune extracerebellar pre-motor structures and induce the learned behavior. PMID- 21645106 TI - Tonic and phasic components of eye movements during REM sleep in the rat. AB - Phasic oculomotor activity is one of the features identifying rapid eye movement (REM) sleep. Recently, it has been shown in cats that, despite bursts of complex two-component rapid eye movements, the eyes tend to maintain a nasal and downward rotation during REM sleep. Although the function of eye movements during sleep remains elusive, it is important to know whether the characteristics of eye movements during sleep are species-specific in mammals. In this work, quantitative characteristics of eye movements, recorded by the scleral search coil technique, were studied during wakefulness and sleep in rats. During wakefulness, rats performed conjugated saccades at a very low rate and some eye movements associated to blinking and gnawing. Throughout non-REM sleep, eye movements were slow, mostly unconjugated and the eyes maintained a divergence in the horizontal plane. The beginning of REM sleep was characterized by a convergence and downward rotation of the eyes, which tended to persist until the end of REM sleep. Rapid eye movements, isolated and monocular at the beginning, became complex and organized in high-frequency bursts. These results demonstrate that, despite the difference in extraocular anatomy and visuomotor strategies between frontal- and lateral-eyed species, eye movements during REM sleep in rats are very similar to those described in cats. This suggests that the mechanisms generating eye movements during REM sleep are largely conserved in mammals. PMID- 21645108 TI - The neural control of fast vs. slow vergence eye movements. AB - When looking between targets located in three-dimensional space, information about relative depth is sent from the visual cortex to the motor control centers in the brainstem, which are responsible for generating appropriate motor commands to move the eyes. Surprisingly, how the neurons in the brainstem use the depth information supplied by the visual cortex to precisely aim each eye on a visual target remains highly controversial. This review will consider the results of recent studies that have focused on determining how individual neurons contribute to realigning gaze when we look between objects located at different depths. In particular, the results of new experiments provide compelling evidence that the majority of saccadic neurons dynamically encode the movement of an individual eye, and show that the time-varying discharge of the saccadic neuron population encodes the drive required to account for vergence facilitation during disconjugate saccades. Notably, these results suggest that an additional input (i.e. from a separate vergence subsystem) is not required to shape the activity of motoneurons during disconjugate saccades. Furthermore, whereas motoneurons drive both fast and slow vergence movements, saccadic neurons discharge only during fast vergence movements, emphasizing the existence of distinct premotor pathways for controlling fast vs. slow vergence. Taken together, these recent findings contradict the traditional view that the brain is circuited with independent pathways for conjugate and vergence control, and thus provide an important new insight into how the brain controls three-dimensional gaze shifts. PMID- 21645107 TI - Binocular coordination of eye movements--Hering's Law of equal innervation or uniocular control? AB - The neurophysiological basis for binocular control of eye movements in primates has been characterized by a scientific controversy that has its origin in the historical conflict of Hering and Helmholtz in the 19th century. This review focuses on two hypotheses, linked to that conflict, that seek to account for binocular coordination - Hering's Law vs. uniocular control of each eye. In an effort to manage the length of the review, the focus is on extracellular single unit studies of premotor eye movement cells and extraocular motoneurons. In the latter half of the 20th century, these studies provided a wealth of neurophysiological data pertaining to the control of vergence and conjugate eye movements. The data were initially supportive of Hering's Law. More recent data, however, have provided support for uniocular control of each eye consistent with Helmholtz's original idea. The controversy is far from resolved. New anatomical descriptions of the disparate inputs to multiply and singly innervated extraocular muscle fibers challenge the concept of a 'final common pathway' as they suggest there may be separate groups of motoneurons involved in vergence and conjugate control of eye position. These data provide a new challenge for interpretation of uniocular premotor control networks and how they cooperate to produce coordinated eye movements. PMID- 21645109 TI - Neck muscle responses evoked by transcranial magnetic stimulation of the human frontal eye fields. AB - Transcranial magnetic stimulation (TMS) provides a non-invasive means of investigating brain function. Whereas TMS of the human frontal eye fields (FEFs) does not induce saccades, electrical stimulation of the monkey FEF evokes eye head gaze shifts, with neck muscle responses evoked at stimulation levels insufficient to evoke a saccade. These animal results motivated us to examine whether TMS of the FEF (TMS-FEF) in humans evokes a neck muscle response. Subjects performed memory-guided saccades to the left or right while TMS (two pulses at 20 Hz) was delivered on 30% of trials to the left FEF coincident with saccade instruction. As reported previously, TMS-FEF decreased contralateral saccade reaction times. We simultaneously recorded the activity of splenius capitis (SPL) (an ipsilateral head turner). TMS-FEF evoked a lateralized increase in the activity of the right SPL but not the left SPL, consistent with the recruitment of a contralateral head-turning synergy. In some subjects, the evoked neck muscle response was time-locked to stimulation, whereas in others the evoked response occurred around the time of the saccade. Importantly, evoked responses were greater when TMS was applied to the FEF engaged in contralateral saccade preparation, with even greater evoked responses preceding shorter latency saccades. These results provide new insights into both the nature of TMS and the human oculomotor system, demonstrating that TMS-FEF engages brainstem oculomotor circuits in a manner consistent with a general role in eye-head gaze orienting. Our results also suggest that pairing neck muscle recordings with TMS-FEF provides a novel way of assaying the covert preparation of oculomotor plans. PMID- 21645110 TI - First two decades of paediatric heart transplantation in Sweden - outcome of listing and post-transplant results. AB - AIMS: To evaluate outcome in the first generation of children with end-stage heart disease to whom heart transplantation was available. METHODS: Retrospective review of all 135 Swedish children <18 years old listed for heart transplantation 1989-2009, followed to December 31, 2009, including 74 (55%) with cardiomyopathy and 61 (45%) with congenital heart disease; 34 (25%) were infants (<1 year). Cumulative risk of requiring heart transplantation was 1:17,300 (11 patients who improved were omitted from outcome analysis). RESULTS: Waiting-list mortality was 31% (44% in infants). Median waiting time in 82 transplanted patients was 57 days (0-585 days). Post-transplant follow-up time was median 5.9 years (0.03-20.1 years), and actuarial survival was 92% at 1 year, 82% at 5 years, 76% at 10 years and 58% at 15 years. Survival after listing was 64% at 1 year, 58% at 5 years, 52% at 10 years and 40% at 15 years. Post-transplant complications included rejections (34%), malignancies (12%), renal failure (8%), coronary artery vasculopathy (6%) and re-transplantation (5%). Among 64 survivors, 84% were free of complications affecting prognosis. CONCLUSION: High waiting-list mortality and post-transplant attrition precluded 60% of this pioneer population from reaching adulthood. Functional status in survivors is generally good. PMID- 21645111 TI - A meta-synthesis of women's postincarceration experiences. AB - OBJECTIVE: To integrate the findings of qualitative studies about the experiences of women reentering the community postincarceration. DATA SOURCES: Qualitative studies were identified using library databases from nursing and other disciplines, including Cumulative Index to Nursing and Allied Health Literature (CINAHL), MedLine, JSTOR, SCOPUS, WilsonWeb, and ProQuest Dissertations and Theses. The keywords women, prisoners, incarceration, community re-integration, re-entry, qualitative research, and narratives were searched in all databases in articles published in the past decade. STUDY SELECTION: Qualitative studies involving women reentering the community postincarceration were eligible for inclusion. Only studies published from 2000 through 2009 were selected to reflect contemporary experiences of women. DATA EXTRACTION AND SYNTHESIS: Using Noblit and Hare's approach, the authors conducted a metasynthesis of 10 qualitative studies. Each study was carefully read with attention to the data as well as the metaphors used by the author(s). The relationships between the studies were synthesized and identified by listing key metaphors, concepts, themes, and/or ideas of each study. The study translations were synthesized into a whole, and the synthesis refined leading to a description of the experience of women reentering the community. CONCLUSION: The following four overarching themes emerged: tenuous transitions, "once a criminal, always a criminal," downward spiral, and tipping points. The themes allow advance practice nurses in women's health to identify the health and psychosocial needs of a vulnerable population of women and to develop interventions that address the challenges women face upon release. PMID- 21645112 TI - Addressing incarcerated women's unique and unidentified health care needs. PMID- 21645113 TI - Toward a gender-responsive restorative correctional health care model. AB - Infectious and chronic diseases are disproportionately greater among detained women than among women in the population at large. The overall punitive, male based, one-size-fits-all approach to health care and the variant overall health care standards across correctional systems call for a new model of correctional health care. The assumptions, constructs, strategies, and future research implications of the restorative health care model are described. PMID- 21645114 TI - End-of-life care and barriers for female inmates. AB - The number of female inmates is growing, and their average age is increasing. As a result, end-of-life care is situated in a highly restrictive environment with a focus on security rather than comfort. We describe the need for and potential barriers to humane care and provide care strategies that can be useful in a complex organizational system. Frontline workers such as nurses who understand the balance between care and control must promote change in the women's prison system. PMID- 21645115 TI - Abuse-related post-traumatic stress, coping, and tobacco use in pregnancy. AB - OBJECTIVE: To examine the relationship between trauma history, posttraumatic stress disorder (PTSD), coping, and smoking in a diverse sample of pregnant women, some of whom are active smokers. DESIGN: Secondary analysis from a prospective study on PTSD and pregnancy outcomes. SETTING: Maternity clinics at three health systems in the midwestern United States. PARTICIPANTS: Women age 18 or older (1,547) interviewed at gestational age fewer than 28 weeks. METHODS: Participants were classified at nonsmokers, quitters (stopped smoking during pregnancy), and pregnancy smokers. Demographic, trauma, and pregnancy factors, substance use, and use of tobacco to cope were compared across groups. Logistic regression assessed the influence of these factors on being a smoker versus a nonsmoker and a quitter versus a pregnancy smoker. RESULTS: Smokers differed from nonsmokers on all demographic risk factors (being African American, being pregnant as a teen, having lower income and less education, and living in high crime areas), had higher rates of current and lifetime PTSD, and were more likely to report abuse as their worst trauma. Pregnancy smokers had lower levels of education, were more likely to classify their worst trauma as "extremely troubling," and were more likely to exhibit PTSD hyperarousal symptoms. In regression models, smoking "to cope with emotions and problems" doubled the odds of continuing to smoke while pregnant even after accounting for several relevant risk factors. CONCLUSION: Smoking behavior in pregnancy may be influenced by the need to cope with abuse-related PTSD symptoms. Clinicians should consider using trauma-informed interventions when working with tobacco-using pregnant women. PMID- 21645116 TI - An overview of distribution of births in United States hospitals in 2008 with implications for small volume perinatal units in rural hospitals. AB - OBJECTIVE: To evaluate the distribution of births among United States (U.S.) hospitals in 2008 as part of the background for the Association of Women's Health, Obstetric and Neonatal Nurses' Guidelines for Professional Registered Nurse Staffing for Perinatal Units. DESIGN: Descriptive analysis of birth volumes in U.S. hospitals using American Hospital Association Annual Survey: 2008. METHODS: U.S. hospitals providing obstetric (OB) services were identified based on information in any of three fields: OB services, OB beds, or number of births. Data were verified via telephone and/or website for the top 100 hospitals based on volume, hospitals with "Healthcare System" as part of their names, hospitals reporting births but no OB service, and hospitals reporting <100 births. Hospitals with <100 births were queried regarding nurse staffing. Descriptive statistics were used to analyze data. RESULTS: Approximately 3,265 U.S. hospitals offered OB services in 2008. The top 500 hospitals based on volume (15.3%) accounted for almost one half (47.4%) of births, the top 1,000 for 69.2%, and the remaining 2,265 for 30.8%. Fourteen percent of hospitals with <100 births in 2008 reported discontinuing OB services in 2010, in part due to lack of physician coverage and costs. Most hospitals (n=159, 72.3%) with <100 births routinely maintained two OB-skilled nurses in-house in 2010. CONCLUSIONS: U.S. births are unevenly distributed among hospitals; 15% have a disproportionate share of nearly one half of all births. Most hospitals (69.4%) are operating medium- to small volume OB units. Most hospitals (72.3%) with <100 births annually reported currently meeting minimum staffing guidelines. PMID- 21645117 TI - A nasally applied cellulose powder in seasonal allergic rhinitis (SAR) in children and adolescents; reduction of symptoms and relation to pollen load. AB - BACKGROUND: A nasally applied cellulose powder is increasingly used in many countries as a remedy for allergic rhinitis. The absence of side effects makes the treatment particularly attractive in children. The efficacy in pollen allergic children, however, is not studied, nor is the relation to various pollen exposures. METHODS: During the birch pollen season in 2009, a double blind, placebo-controlled study was conducted in 53 subjects, aged 8-18 yr, with allergic rhinitis attributed to birch pollen. All children were on daily oral antihistamine. Reminders and reporting of symptom scores were made by SMS on mobile phones. Pollen was collected in a volumetric trap from which figures of pollen concentrations from 1979 to 2009 were available. RESULTS: There was a significant reduction in total symptom scores from the nose (Placebo 7.29, Active 6.07, p = 0.033) and specifically for running nose (Placebo 2.56, Active 2.03, p = 0.017). All symptoms from the nose, eyes and lower airways were lower in the active group but reached significance only as earlier. The best effect was seen after days with low or moderate pollen counts (<=100/m(3)), the predominating pollen load over 31 yr in the area. No clinically significant adverse effects were seen. CONCLUSIONS: The product reduces symptoms of SAR in children and adolescents. Original data on pollen concentrations over 31 yr are presented with levels mainly in the low range favouring the observed efficacy profile. SMS communication on mobile phone for reminders and recording symptom scores was an excellent logistics tool. PMID- 21645118 TI - Educational case series: Mechanisms of drug allergy. AB - Once administered, a drug can activate the immune system by various mechanisms and lead to a large range of clinical manifestations closely related to the type of immune reaction elicited. Administration of the drug can classically result in an immunoglobulin E (IgE)-type sensitization, but can also result in more complex activation of the immune system potentially resulting in severe syndromes, such as the drug-induced hypersensitivity syndrome (DIHS). Although there has been a major increase in our knowledge over the last years, the exact mechanisms of drug allergy are not well understood for most clinical manifestations. A complex interaction between individual characteristics, environmental factors, and the drug itself is usually responsible for adverse reactions to drugs. In this educational review series, we described three cases of drug allergy: first, a child with a typical IgE-mediated drug allergy, second, a child with a non immediate reaction to penicillin, and in the third patient, we will discuss the drug-induced hypersensitivity syndrome, which is rare but potentially fatal. These cases are correlated to the immune mechanism potentially involved. PMID- 21645119 TI - Osteoporosis: an ignored complication of CVID. AB - BACKGROUND: Multiple factors in common variable immunodeficiency (CVID) might interfere with optimal growth and maturation and potentially compromise bone health. METHODS: We aimed to evaluate bone mineral density (BMD) of patients with CVID using dual energy X-ray absorptiometry (DEXA) and investigate risk factors associated with decreased bone density. RESULTS: Twenty-two patients were included (M: 16, F: 6) with a mean age of 15.6 +/- 9.0 yr. DEXA revealed osteopenia in 6/22 (27.3%) and osteoporosis in 9/22 (40.9%) at lumbar spine and osteopenia in 7/19 (37%) and osteoporosis in 3/19 (16%) at femoral neck sites. The age of subjects with osteoporosis was significantly higher than those without (21.6 +/- 8.0 vs. 9.0 +/- 5.7 yr; p < 0.0001). BMD z-scores were significantly lower in patients with bronchiectasis compared with those without (p = 0.03). Patients with osteoporosis at femoral neck site had lower forced expiratory volume in 1 s (FEV(1) ) (p = 0.024), FEV(1) /forced vital capacity (FVC) (p < 0.0001), PEF (p = 0.008), and FEF 25-75 (p = 0.013) values compared with the patients with normal BMD z-scores. Low serum 25(OH) vitamin D levels were detected in 13/22 patients and low dietary calcium intake in 17/22 patients. BMD z-scores at femoral neck were lower in patients with low B-cell percentage (p = 0.03). BMD z-score at lumbar spine was correlated with folate (r = +0.63, p = 0.004) and serum immunoglobulin G levels (r = +0.430, p = 0.04). CONCLUSION: Osteoporosis appeared as an emerging health problem of patients with CVID, the risk increasing with older age and poorer lung function. Nutritional, biochemical, and immunologic factors appeared to take part in decreased BMD. Insight into the mechanisms of osteoporosis in CVID is crucial to develop preventive strategies. PMID- 21645120 TI - Flap extension attained by vertical and periosteal-releasing incisions: a prospective cohort study. AB - OBJECTIVES: To evaluate the effect of vertical and periosteal-releasing incisions (PRI) on the extension of the buccal flap in a trapezoidal flap design. MATERIAL AND METHODS: Thirty patients scheduled for implant surgery accompanied by bone augmentation were recruited. The amount of flap extension pulled with a minimal tension of 5 g using a dynamometer was measured before and after the first and the second vertical incisions (VI) and PRI. The results were compared based on gender, surgical site and operator's experience. RESULTS: The first VI extended flap length by 1.1 +/- 0.6 mm, 113.4% more compared with the original flap length. The second VI increased flap length by 1.9 +/- 1 mm (124.2%), and the PRI significantly extended flap length by 5.5 +/- 1.5 mm (171.3%) (P<0.001). The length of the first and the second VI and the amount of flap extension by each incision were compared, and there were no statistically significant differences between gender, surgical site (maxilla vs. mandible) and operator's experience (faculty vs. residents). CONCLUSION: Vertical and PRI in a trapezoidal flap design can be successfully utilized to attain tension-free primary closure during implant or periodontal surgeries. However, the PRI appears to be the only determinant key factor that can significantly extend the length of the flaps, while host- and operator-related factors might not have any significant effect on such flap extension. PMID- 21645121 TI - Hypopyon-like features: new dermoscopic criteria in the differential diagnosis of cutaneous lymphangioma circumscriptum and haemangiomas? AB - BACKGROUND: Lymphangiomas are congenital malformations of the lymphatic system characterized by clusters of translucent vesicles. Dermoscopically, yellow and bluish lacunea according to the lymphatic or haematic content were described previously, but it is difficult to distinguish them from haemangioma. OBSERVATIONS: In nine patients with cutaneous lymphangioma circumscriptum (CLC), hypopyon-like features were observed. None of the 10 patients with haemangioma had this finding. CONCLUSION: Hypopyon-like features can be a useful dermoscopic pattern in differentiating CLC from haemangioma. PMID- 21645122 TI - The diagnosis of early psoriatic arthritis in an outpatient dermatological centre for psoriasis. AB - BACKGROUND: Most of the data currently available on early psoriatic arthritis (EPsA) derive from studies performed in rheumatological settings. However, in recent years, there has been an increase in the amount of data from dermatologic centres. OBJECTIVES: To describe the prevalence, clinical, laboratory and imaging characteristics of psoriatic patients with EPsA seen at a dermatological outpatient psoriasis centre. METHODS: From January 2007 to May 2010, all patients with psoriasis who visited the psoriasis centre were asked about inflammatory joint involvement. A diagnosis of psoriatic arthritis was made on the basis of clinical, laboratory and imaging studies. The patients were diagnosed with early PsA (EPsA) if their inflammatory articular symptoms had been present for <= 1 year. RESULTS: We diagnosed EPsA in 33 patients. Joint involvement was polyarticular (>5 joints involved) in 20 patients (60.6%) and oligoarticular (<=5 joints involved) in the remaining 13 patients. Quality of life due to skin involvement and the degree of functional impairment due to joint inflammation were only mildly affected, as measured by DLQI and HAQ, respectively. A direct correlation between the number of tender joints (ACR 68) and HAQ was found (r = 0.36; P = 0.04). Imaging studies showed that in spite of the absence of radiologic findings of peripheral joint damage, ultrasonography and contrast enhanced ultrasonography showed signs of articular inflammation in all patients. CONCLUSIONS: A diagnosis of EPsA can be correctly performed in a dermatologic outpatient facility. To do so, a close collaboration among dermatologists, rheumatologists and radiologists is necessary. PMID- 21645123 TI - Interest of external photoprotection by means of clothing and sunscreen products in young children. AB - BACKGROUND: Protecting young children is a major public health issue. OBJECTIVE: We tested children's clothing and sunscreen products that are specially designed for children. METHODS: The effectiveness of the different types of clothing and sunscreen products was determined using spectrophotometer equipped with an integrating sphere A measure of transmittance carried out, respectively, between 290 and 400 nm and between 320 and 400 nm, allowed us to assess the photoprotective properties in the UVB and UVA range respectively. RESULTS: Although the photoprotective effect varies according to the type of clothing (UV protection factor [UPF] of approximately 10 for a cotton T-shirt and 500 for a pair of jeans), the effect is constant for any one type of clothing. Placing fabrics in layers is essential and this enables the protective effect to be greatly increased. Moreover, the authors wished to answer the question, 'what makes a good sun cream?' by analysing the formula of each product tested. It turns out that only creams containing both organic filters and inorganic filters have an sun protection factor (SPF) that is the same as the one stated on the product. CONCLUSION: Concerning townwear, the jeans, tracksuits, sweatshirts, pullovers and tights turned out to be very photoprotective. They enable, in effect, to reach a UPF higher than 500. The mere presence of titanium dioxide and/or zinc dioxide does not mean that the product will have a high SPF. Products containing alcohol can be eliminated from the possible choices by carefully reading the label. PMID- 21645124 TI - Management of dermatofibrosarcoma protuberans with fibrosarcomatous transformation: an evidence-based review of the literature. AB - Fibrosarcomatous transformation represents a rare event in dermatofibrosarcoma protuberans (DFSP) with unpredictable biological behaviour. No guidelines for the adequate treatment of patients with this rare neoplasm have been published. Herein, we present a comprehensive review of the literature comprising 157 patients with transformed DFSP focussing on surgical and adjuvant treatment modalities for this tumour. In the cohort examined, local recurrence occurred in 36% of cases and was significantly lower in patients treated by wide excision with margins >=2 cm when compared with those treated with local excision without defined margins (P = 0.01). Consistently, negative margin status was associated with a lower recurrence rate when compared with positive or unknown margin status (P = 0.01). Distant metastases were detected in 13% of patients, which is significantly higher when compared with ordinary dermatofibrosarcoma protuberans. Systemic dissemination was preceded by local recurrence in 81% of cases, and is therefore strongly associated with tumour recurrence (P <= 0.001). The present data confirm that wide excision with margins >= 2 cm represent the gold standard in the treatment of transformed dermatofibrosarcoma protuberans, and prevents recurrence as well as metastasis. When R0-resection is not feasible, adjuvant radiation should be considered for cases with incomplete resection or unknown surgical margins. Irresectable or metastatic transformed DFSP harbouring the COL1A1-PDGFB fusion gene should be treated with imatinib in the palliative setting or as an adjunctive treatment before surgery, although responses may be short-lasting. PMID- 21645125 TI - Tapering cyclosporine after long-term treatment of atopic dermatitis. PMID- 21645126 TI - Canine mammary tumours, an overview. AB - Canine mammary tumours (CMTs) are the most common neoplasms in intact female dogs. Although the prevalence of these tumours decreases in regions where preventive ovari(ohyster)ectomy is performed, it remains an important disease entity in veterinary medicine. Moreover, treatment options are limited in comparison with human breast cancer. Nevertheless, recent human treatment protocols might have potential in bitches suffering from CMTs. PMID- 21645127 TI - Interactions between straw size and thawing rates on the cryopreservation of agouti (Dasyprocta aguti) epididymal sperm. AB - This study verifies the interactions between straw size and thawing rates and their impact on the epididymal sperm from this species. Caudae epididymidum from 10 agoutis were subjected to retrograde washing using a coconut water extender (ACP-109c((r)) ). Epididymal sperm were evaluated and extended in ACP-109c((r)) plus egg yolk (20%) and glycerol (6%). The samples were packaged in 0.25- or 0.50 ml straws, frozen in liquid nitrogen and thawed at 37 degrees C/1 min or 70 degrees C/8 s, followed by a re-evaluation. The use of 0.25-ml straws thawed at 37 degrees C/1 min provided a value of 26.6% for sperm motility. No interactions between straw size and thawing rates were verified on agouti sperm (p > 0.05), but when 0.5-ml straws were thawed at 70 degrees C/8 s, sperm vigour decreased significantly (p < 0.05). It is recommended that the agouti epididymal sperm cryopreserved in ACP-109c((r)) extender should be packaged in 0.25- or 0.50-ml straws and thawed at 37 degrees C/60 s. PMID- 21645128 TI - Semen traits and seminal plasma biochemical parameters in white leghorn layer breeders. AB - This study was designed to determine the semen quality and seminal plasma biochemical parameters in White Leghorn layer breeders in the early phase of maturity. Individual ejaculates from 25 males were analysed for the determination of volume, sperm concentration, dead sperm percentage (DS) and sperm motility. Seminal plasma was separated and analysed for total oxidant status (TOS), total antioxidant capacity (TAC), paraoxonase (PON1), arylesterase, ceruloplasmin, triiodothyronine, thyroxine, aspartate aminotransferase (AST) and alanine aminotransferase (ALT). Our findings showed a significant (p < 0.05) positive correlation between TOS and DS in layer breeders. The values of TAC were significantly negatively correlated with TOS and DS, while positively correlated with PON1. Conversely, AST showed significant negative correlation with motility and DS. Significantly, negative correlation was also observed between ALT and sperm concentration. In conclusion, these parameters provide some valuable basal data that may help better understanding the semen quality characteristics of White Leghorn layer breeders. PMID- 21645129 TI - Exocytosis and growth do not occur only at hyphal tips. AB - Mycologists have put extreme emphasis on hyphal tip growth as the primary mode of growth in filamentous fungi. Much attention has also been focused on the exocytosis of extracellular enzymes from hyphal tips. However, growth and exocytosis commonly occur at hyphal locations other than tips. Here I briefly review our limited understanding of growth and exocytosis during intercalary hyphal extension, subapical branch initiation, septum formation and secondary wall thickening. Secretion of extracellular enzymes and adhesion molecules from subapical hyphal regions is also discussed. Recent research using advanced live cell imaging techniques (e.g. Hayakawa et al., 2011 in this issue) is providing new insights into the mechanistic basis of many of these processes. PMID- 21645130 TI - Functional characterization of the ferroxidase, permease high-affinity iron transport complex from Candida albicans. AB - Saccharomyces cerevisiae expresses two proteins that together support high affinity Fe-uptake. These are a multicopper oxidase, Fet3p, with specificity towards Fe2+ and a ferric iron permease, Ftr1p, which supports Fe-accumulation. Homologues of the genes encoding these two proteins are found in all fungal genomes including those for the pathogens, Candida albicans and Cryptococcus neoformans. At least one of these loci represents a virulence factor for each pathogen suggesting that this complex would be an appropriate pharmacologic target. However, the mechanism by which this protein pair supports Fe-uptake in any fungal pathogen has not been elucidated. Taking advantage of the robust molecular genetics available in S. cerevisiae, we identify the two of five candidate ferroxidases likely involved in high-affinity Fe-uptake in C. albicans, Fet31 and Fet34. Both localize to the yeast plasma membrane and both support Fe uptake along with an Ftr1 protein, either from C. albicans or from S. cerevisiae. We express and characterize Fet34, demonstrating that it is functionally homologous to ScFet3p. Using S. cerevisiae as host for the functional expression of the C. albicans Fe-uptake proteins, we demonstrate that they support a mechanism of Fe-trafficking that involves channelling of the CaFet34-generated Fe3+ directly to CaFtr1 for transport into the cytoplasm. PMID- 21645131 TI - CsgE is a curli secretion specificity factor that prevents amyloid fibre aggregation. AB - Curli are extracellular amyloid fibres produced by Escherichia coli that are critical for biofilm formation and adhesion to biotic and abiotic surfaces. CsgA and CsgB are the major and minor curli subunits, respectively, while CsgE, CsgF and CsgG direct the extracellular localization and assembly of curli subunits into fibres. The secretion and stability of CsgA and CsgB are dependent on the outer membrane lipoprotein CsgG. Here, we identified functional interactions between CsgG and CsgE during curli secretion. We discovered that CsgG overexpression restored curli production to a csgE strain under curli-inducing conditions. In antibiotic sensitivity and protein secretion assays, CsgG expression alone allowed translocation of erythromycin and small periplasmic proteins across the outer membrane. Coexpression of CsgE with CsgG blocked non specific protein and antibiotic passage across the outer membrane. However, CsgE did not block secretion of proteins containing a 22-amino-acid putative outer membrane secretion signal of CsgA (A22). Finally, using purified proteins, we found that CsgE prohibited the self-assembly of CsgA into amyloid fibres. Collectively, these data indicate that CsgE provides substrate specificity to the curli secretion pore CsgG, and acts directly on the secretion substrate CsgA to prevent premature subunit assembly. PMID- 21645133 TI - Impact of expected value on neural activity in rat substantia nigra pars reticulata. AB - The substantia nigra pars reticulata (SNr) is thought to serve as the output of the basal ganglia, whereby associative information from striatum influences behavior via disinhibition of downstream motor areas to motivate behavior. Unfortunately, few studies have examined activity in SNr in rats making decisions based on the value of predicted reward similar to those conducted in primates. To fill this void, we recorded from single neurons in SNr while rats performed a choice task in which different odor cues indicated what reward was available on the left or on the right. The value of reward associated with a leftward or rightward movement was manipulated by varying the size of and delay to reward in separate blocks of trials. Rats were faster or slower depending on whether the expected reward value was high or low, respectively. The number of neurons that increased firing during performance of the task outnumbered those that decreased firing. Both increases and decreases were modulated by expected value and response direction. Neurons that fired more or less strongly for larger reward tended to fire, respectively, more or less strongly for immediate reward, reflecting their common motivational output. Finally, value selectivity was present prior to presentation of cues indicating the nature of the upcoming behavioral response for both increasing- and decreasing-type neurons, reflecting the internal bias or preparatory set of the rat. These results emphasize the importance of increasing-type neurons on behavioral output when animals are making decisions based on predicted reward value. PMID- 21645134 TI - Patterned Purkinje cell loss in the ataxic sticky mouse. AB - The ataxic sticky (sti/sti) mouse is a spontaneous autosomal recessive mutant resulting from a disruption in the editing domain of the alanyl-tRNA synthetase (Aars) gene. The sticky phenotype is characterized by a small body size, a characteristic unkempt coat and neurological manifestations including marked tremor and ataxia starting at 6 weeks of age. The present study was undertaken to examine the spatiotemporal features of Purkinje cell degeneration in the sticky mouse. Purkinje cell loss was found to be both progressive and patterned, with vermal lobules VI, IX and X, crus 1 of the hemisphere, and the flocculus and paraflocculus being differentially resistant to degeneration. The pattern of Purkinje cell degeneration in sticky is not random - in general, the sphingosine kinase 1a-immunonegative Purkinje cell subset is preferentially susceptible to early cell death. In addition, zebrin II/aldolase C expression in the sticky cerebellum is profoundly downregulated, whereas the heat-shock protein 25 is both ectopically expressed in some scattered Purkinje cells and downregulated in other Purkinje cells in which it is normally expressed constitutively. Compared with many mouse mutants with patterned Purkinje cell death, in which successive stripes of cell loss are very clear, Purkinje cell loss in sticky shows a less clear-cut pattern between different Purkinje cell subtypes, with the result that preferential survival is less dramatic. This may represent a secondary consequence of the downregulation of zebrin II expression. PMID- 21645132 TI - Niche-specific contribution to streptococcal virulence of a MalR-regulated carbohydrate binding protein. AB - Low G+C Gram-positive bacteria typically contain multiple LacI/GalR regulator family members, which often have highly similar amino-terminal DNA binding domains, suggesting significant overlap in target DNA sequences. The LacI/GalR family regulator catabolite control protein A (CcpA) is a global regulator of the Group A Streptococcus (GAS) transcriptome and contributes to GAS virulence in diverse infection sites. Herein, we studied the role of the maltose repressor (MalR), another LacI/GalR family member, in GAS global gene expression and virulence. MalR inactivation reduced GAS colonization of the mouse oropharynx but did not detrimentally affect invasive infection. The MalR transcriptome was limited to only 25 genes, and a highly conserved MalR DNA-binding sequence was identified. Variation of the MalR binding sequence significantly reduced MalR binding in vitro. In contrast, CcpA bound to the same DNA sequences as MalR but tolerated variation in the promoter sequences with minimal change in binding affinity. Inactivation of pulA, a MalR regulated gene which encodes a cell surface carbohydrate binding protein, significantly reduced GAS human epithelial cell adhesion and mouse oropharyngeal colonization but did not affect GAS invasive disease. These data delineate a molecular mechanism by which hierarchical regulation of carbon source utilization influences bacterial pathogenesis in a site-specific fashion. PMID- 21645135 TI - Improvements in rate of development and magnitude of force with intense auditory stimuli in patients with Parkinson's disease. AB - Patients with Parkinson's disease can show brief but dramatic normalization of motor activity in highly arousing situations, a phenomenon often termed paradoxical kinesis. We sought to mimic this in a controlled experimental environment. Nine patients with Parkinson's disease and nine age-matched healthy controls were asked to grip a force dynamometer as quickly and strongly as possible in response to a visual cue. A loud (96 dB) auditory stimulus was delivered at the same time as the visual cue in ~50% of randomly selected trials. In patients with Parkinson's disease, the experiment was conducted after overnight withdrawal of antiparkinsonian drugs and again 1 h after patients had taken their usual morning medication. Patients showed improvements in the peak rate of force development and the magnitude of force developed when loud auditory stimuli accompanied visual cues. Equally, they showed improvements in the times taken to reach the peak rate of force development and their maximal force. The paradoxical facilitatory effect of sound was similar whether patients were off or on their usual antiparkinsonian medication, and could be reproduced in age matched healthy controls. We conclude that motor improvement induced by loud auditory stimuli in Parkinson's disease is related to a physiological phenomenon which survives both with and after withdrawal of antiparkinsonian medication. The potential independence of the mediating pathways from the dopaminergic system provides impetus for further investigation as it may yield a novel nondopaminergic target for therapeutic manipulation in Parkinson's disease. PMID- 21645136 TI - Persistent activity in layer 5 pyramidal neurons following cholinergic activation of mouse primary cortices. AB - Persistent spiking activity is thought to be a cellular process involved in working memory. We have been interested in whether persistent activity also exists in cortical areas which are not involved in this memory process. To study the possible presence and the mechanisms of persistent activity in layer 5 pyramidal cells of the mouse primary somatosensory, visual and motor cortices, we used patch-clamp and calcium imaging techniques. A combination of cholinergic receptor activation and suprathreshold depolarization or sufficient extracellular stimulation leads to either a subthreshold afterdepolarization or suprathreshold persistent activity in these cortices. There is a continuum of response amplitudes depending on depolarization size. To initiate persistent activity, spikes have to be induced at a frequency of at least 20 Hz, if tested for 1 s. Acetylcholine muscarinic, but not nicotinic, receptors are important for initiating persistent activity. Persistent activity is an intrinsic cellular, not a network, phenomenon as it persists under blockade of ionotropic glutamate and GABA receptors. A rise in intracellular calcium concentration through voltage gated calcium channels is needed for persistent activity initiation, while intracellular calcium stores are not crucial. The increased intracellular calcium concentration leads to the activation of calcium-sensitive nonspecific cationic channels. This study for the first time describes the presence and the underlying mechanisms of persistent activity in pyramidal cells of three primary sensory and motor cortex areas. These results thereby suggest that persistent activity may be a general capability of deep layer cortical pyramidal cells. PMID- 21645137 TI - Plasma IL-6 concentration correlates with clinical disease activity and serum C reactive protein concentration in chronic urticaria patients. AB - BACKGROUND: Our previous study was the first to demonstrate enhanced plasma IL-6 concentrations in chronic urticaria (CU). It is known that C-reactive protein (CRP) is a sensitive marker of an underlying systemic inflammation, triggered mainly as a response to IL-6. OBJECTIVE: To evaluate plasma IL-6 concentration in CU patients relating to the clinical disease activity and serum CRP concentration. METHODS: Serum CRP and plasma IL-6 concentrations were measured in 58 CU patients and 30 healthy subjects. Ten CU patients were evaluated twice, during the active period as well as upon the spontaneous clinical remission of the disease. CU activity was assessed with the use of the symptom scores recommended by EAACI/GALEN/EDF guidelines. RESULTS: IL-6 and CRP concentrations were significantly increased in CU patients as compared with the healthy subjects, whereas they decreased remarkably upon the spontaneous remission. IL-6 concentration was associated with weekly urticaria activity scores and also significant differences were found between patients showing different degrees of urticarial activity. Significant correlation was observed between IL-6 and CRP concentrations. CONCLUSIONS AND CLINICAL RELEVANCE: This study reinforces evidence that, apart from a local cutaneous inflammation, CU is associated with a systemic inflammatory response. Such acute-phase response is manifested by increased circulating IL-6, which varies along with CRP changes and may be related to the urticarial activity. PMID- 21645139 TI - Oncocytic variant of papillary thyroid carcinoma with lymphocytic stroma (Warthin like variant): report of a case with fine needle aspiration cytology and review of the literature. PMID- 21645140 TI - Persistent, widespread papilloma formation on the penis of a horse: a novel presentation of equine papillomavirus type 2 infection. AB - A 9-year-old gelding presented with approximately 100 papillomas that covered about 75% of the distal penis. Biopsy was performed, and histology showed evidence of viral cytopathic change and koilocytosis. Polymerase chain reaction using DNA extracted from biopsied tissue amplified equine papillomavirus type 2 (EcPV-2) DNA sequences. Sixteen months later, the horse was re-examined and the appearance of the papillomas was unchanged. Equine papillomavirus type 2 DNA sequences were again amplified from both biopsied tissue and swabs of the penis. Papillomavirus was localized to the lesions by immunohistochemistry and in situ hybridization. An examination 2 years after the initial presentation revealed no detectable change in the appearance of the penis. The large number of papillomas and their failure to regress over an extended period support a clinical classification of papillomatosis. To the authors' knowledge, this is the first report of papillomatosis of the equine penis. This novel clinical manifestation suggests that persistent EcPV-2 infection is possible in horses. As there is evidence that EcPV-2 may promote development of equine penile squamous cell carcinoma, understanding the natural history of EcPV-2 infections may be important in preventing equine penile neoplasia. PMID- 21645138 TI - Basic mechanisms of itch. AB - Chronic itch represents a burdensome clinical problem that can originate from a variety of aetiologies. Pruriceptive itch originates following the activation of peripheral sensory nerve endings following damage or exposure to inflammatory mediators and ascends to the brain through the spinal thalamic tract. Much insight has been gained into the understanding of the mechanisms underlying pruriceptive itch through studies using humans and experimental animals. More than one sensory nerve subtype is thought to subserve pruriceptive itch which includes both unmyelinated C-fibres and thinly myelinated Adelta nerve fibres. There are a myriad of mediators capable of stimulating these afferent nerves leading to itch, including biogenic amines, proteases, cytokines, and peptides. Some of these mediators can also evoke sensations of pain and the sensory processing underlying both sensations overlaps in complex ways. Studies have demonstrated that both peripheral and central sensitization to pruritogenic stimuli occur during chronic itch. PMID- 21645141 TI - Bacterial contamination of commercial ear cleaners following routine home use. AB - Ear cleaning solutions are designed for repeated use, which raises the possibility for bacterial contamination leading to recurrent or persistent infectious otitis. The purpose of this study was to investigate the prevalence of bacterial contamination of commercial ear cleaners following routine home use in dogs and to describe the characteristics that are associated with contamination. Used ear cleaner bottles and information regarding their use were obtained from canine owners visiting veterinary dermatologists. Both the bottle applicator tips and the solution contents were cultured for aerobic bacteria. Bacterial contamination was present on 10% of the bottle tips and in 2% of the solutions. Isolated bacteria included Staphylococcus pseudintermedius, Bacillus spp., coagulase-negative Staphylococcus spp., Micrococcus spp. and Burkholderia cepacia. The contamination rate was significantly higher on the applicator tips than in the solutions (P = 0.0076). The applicator tip contamination rate was significantly higher in expired samples (17%) than in-date samples (4%; P = 0.0277). The bottle sizes were significantly larger for the samples with contaminated applicator tips compared with noncontaminated tips (P = 0.0455). The contamination rate was significantly higher when Tris-EDTA was an ingredient. Cleanliness of the bottle, contact with the ear canal and infection status of the ear at time of culture had no bearing on the contamination rate. In summary, with routine home use of commercial ear cleaners, pathogenic bacterial contamination is of minor concern. This concern may increase when expired products or larger bottles of ear cleaner are used and when Tris-EDTA is an ingredient. PMID- 21645142 TI - Guidelines on the use of systemic aminoglycosides in veterinary dermatology. PMID- 21645143 TI - Lipid metabolism abnormalities in alcohol-treated rabbits: a morphometric and haematologic study comparing high and low alcohol doses. AB - The pathogenesis of alcohol-induced osteonecrosis remains unclear. The purpose of the present study was to evaluate the morphological changes in bone marrow fat cells and the changes in the serum lipid levels in alcohol-treated rabbits. Fifteen rabbits were randomly assigned into three groups: Four rabbits intragastrically received low-dose alcohol (LDA) (15 ml/kg per day) containing 15% ethanol for 4 weeks, five rabbits received high-dose alcohol (HDA) (30 ml/kg per day) for 4 weeks and six rabbits received physiologic saline for 4 weeks as a control group. Six weeks after the initial alcohol administration, all rabbits were sacrificed. The mean size of the bone marrow fat cells in rabbits treated with HDA was significantly larger than that in the control group (P = 0.0001). Haematologically, the levels of triglycerides and free fatty acids in the rabbits treated with both low-dose and HDA were significantly higher than those in the control group (P = 0.001 for both comparisons). The results of this study are that there are lipid metabolism abnormalities, both morphologically and haematologically, after alcohol administration. Also these findings were more apparent in rabbits treated with HDA than those treated with LDA. PMID- 21645144 TI - Platelet-derived growth factor and platelet-derived growth factor receptor-alpha expression in the normal human thymus and thymoma. AB - Platelet-derived growth factor (PDGF) and its receptors (PDGFRs) are strongly involved in the normal development of several organs, tumour angiogenesis and malignant progression and metastasis. Few studies concerning their expression, distribution and role in normal and pathological human thymus are available in the literature. The aim of this study has been to analyse the immunohistochemical expression of PDGF and PDGFR-alpha in prenatal and postnatal normal human thymus and thymomal biopsy specimens. The results demonstrated immunoreactivity to both PDGF and PDGFR-alpha in all specimens, but the intensity, distribution and number of positive cells were different in normal thymus and thymomas, and also among different tumour types. PDGF and PDGFR-alpha were weakly expressed in foetal and postnatal humans with a different distribution between cortex and medulla in both blood vessels and epithelial cells, whereas they were overexpressed in thymoma, especially in type B2 and B3, in the tumour epithelial cells. Overall, these data suggest that PDGF and PDGFR-alpha may be involved in the pathophysiology of the human thymus. PMID- 21645145 TI - The occurrence of 'bulbs', a complex configuration of the vacuolar membrane, is affected by mutations of vacuolar SNARE and phospholipase in Arabidopsis. AB - The plant vacuole fulfills a variety of functions, and is essential for plant growth and development. We previously identified complex and mobile structures on the continuous vacuolar membrane, which we refer to as 'bulbs'. To ascertain their biological significance and function, we searched for markers associated with bulbs, and mutants that show abnormalities with respect to bulbs. We observed bulb-like structures after expression of non-membranous proteins as well as the functional soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) molecules VAM3 and VTI11. Bulbs are formed in more tissues than previously reported, including flowering organs, suspension culture cells, endodermal cells in the flowering stem, and at very early stages of seed germination. Using existing and newly developed marker lines, we found that the frequency of bulb occurrence is significantly decreased in multiple shoot gravitropism (sgr) mutants, which are known to have a defect in vacuolar membrane properties in endodermal cells. Based on results with new marker lines, which enabled us to observe the process of bulb biogenesis, and analysis of the phenotypes of these mutants, we propose multiple mechanisms for bulb formation, one of which may be that used for formation of transvacuolar strands. PMID- 21645146 TI - Autoimmune response and repression of mitotic cell division occur in inter specific crosses between tetraploid wheat and Aegilops tauschii Coss. that show low temperature-induced hybrid necrosis. AB - Common wheat is an allohexaploid species originating from a naturally occurring inter-specific cross between tetraploid wheat and the diploid wild wheat Aegilops tauschii Coss. Artificial allopolyploidization can produce synthetic hexaploid wheat. However, synthetic triploid hybrids show four types of hybrid growth abnormalities: type II and III hybrid necrosis, hybrid chlorosis, and severe growth abortion. Of these hybrid abnormalities, type II necrosis is induced by low temperature. Under low temperature, elongation of stems and expansion of new leaves is repressed in type II necrosis lines, which later exhibit necrotic symptoms. Here, we characterize type II necrosis in detail. Comparative transcriptome analysis showed that a number of defense-related genes were highly up-regulated in seedling leaves that showed type II necrosis. Transmission electron microscopy revealed extensive cell death in the leaves under low temperature conditions, accompanied by abundant generation of reactive oxygen species. In addition, down-regulation of cell cycle-related genes was observed in shoot apices of type II necrosis lines under low-temperature conditions. Quantitative RT-PCR and in situ hybridization showed repression of accumulation of histone H4 transcripts in the shoot apical meristem of type II necrosis lines. These results strongly suggest that an autoimmune response-like reaction and repression of cell division in the shoot apical meristem are associated with the abnormal growth phenotype in type II necrosis lines. PMID- 21645147 TI - Type-A response regulators are required for proper root apical meristem function through post-transcriptional regulation of PIN auxin efflux carriers. AB - The phytohormones cytokinin and auxin regulate a diverse array of plant processes, often acting together to modulate growth and development. Although much has been learned with regard to how each of these hormones act individually, we are just beginning to understand how these signals interact to achieve an integrated response. Previous studies indicated that exogenous cytokinin has an effect on the transcription of several PIN efflux carriers. Here we show that disruption of type-A Arabidopsis response regulators (ARRs), which are negative regulators of cytokinin signalling, alters the levels of PIN proteins and results in increased sensitivity to N-1-naphthylphthalamic acid, an inhibitor of polar auxin transport. Disruption of eight of the 10 type-A ARR genes affects root development by altering the size of the apical meristem. Furthermore, we show that the effect of cytokinin on PIN abundance occurs primarily at the post transcriptional level. Alterations of PIN levels in the type-A ARR mutants result in changes in the distribution of auxin in root tips as measured by a DR5::GFP reporter, and an altered pattern of cell division and differentiation in the stem cell niche in the root apical meristem. Together, these data indicate that cytokinin, acting through the type-A ARRs, alters the level of several PIN efflux carriers, and thus regulates the distribution of auxin within the root tip. PMID- 21645148 TI - ATG2, an autophagy-related protein, negatively affects powdery mildew resistance and mildew-induced cell death in Arabidopsis. AB - The molecular interactions between Arabidopsis and the pathogenic powdery mildew Golovinomyces cichoracearum were studied by characterizing a disease-resistant Arabidopsis mutant atg2-2. The atg2-2 mutant showed enhanced resistance to powdery mildew and dramatic mildew-induced cell death as well as early senescence phenotypes in the absence of pathogens. Defense-related genes were constitutively activated in atg2-2. In atg2-2 mutants, spontaneous cell death, early senescence and disease resistance required the salicylic acid (SA) pathway, but interestingly, mildew-induced cell death was not fully suppressed by inactivation of SA signaling. Thus, cell death could be uncoupled from disease resistance, suggesting that cell death is not sufficient for resistance to powdery mildew. ATG2 encodes autophagy-related 2, a protein known to be involved in the early steps of autophagosome biogenesis. The atg2-2 mutant exhibited typical autophagy defects in autophagosome formation. Furthermore, mutations in several other ATG genes, including ATG5, ATG7 and ATG10, exhibited similar powdery mildew resistance and mildew-induced cell death phenotypes. Taken together, our findings provide insights into the role of autophagy in cell death and disease resistance, and may indicate general links between autophagy, senescence, programmed cell death and defense responses in plants. PMID- 21645149 TI - The ethylene response factor AtERF11 that is transcriptionally modulated by the bZIP transcription factor HY5 is a crucial repressor for ethylene biosynthesis in Arabidopsis. AB - The phytohormones abscisic acid (ABA) and ethylene are known to play multiple roles in plant development and stress responses. Ethylene biosynthesis is affected by several factors including drought, cold and the phytohormone auxin, although the role of ABA is unclear. In this work ABA-responsive mutants were screened and a bZIP transcription factor HY5 was identified as a negative regulator of ethylene biosynthesis via modulation of the expression of the ethylene biosynthesis genes ACS2 and ACS5. Members of the ethylene response factor (ERF) family of transcriptional repressors in Arabidopsis have been shown to modulate ABA responses and three ERF members were found to carry putative HY5 binding cis-acting elements. Analyses with biochemical and molecular approaches revealed that HY5 specifically binds to the G-box region of the AtERF11 promoter to activate its transcription. We further demonstrate that AtERF11, which contains a repressor motif at its C-terminal, interacts with the dehydration responsive element in the ACS2/5 promoters, to repress its expression, resulting in decreased ethylene biosynthesis. Moreover, an AtERF11 knockout mutant showed increased levels of ACS2/5 expression and ethylene emission, while treatment with ABA greatly suppressed ACS5 transcripts but not ACS2 expression and the ethylene content, indicating that AtERF11 is a key negative regulator for ABA-mediated control of ethylene synthesis. In addition, in ethylene over-producer mutants, ABA treatment was shown to suppress ACS5 transcripts and ethylene content, thereby affecting growth and development. Based on these data, in this research we present a model suggesting that the HY5-AtERF11 regulon is a key factor modulating ABA-regulated ethylene biosynthesis. PMID- 21645150 TI - CXCL13 mediates prostate cancer cell proliferation through JNK signalling and invasion through ERK activation. AB - OBJECTIVES: The focus of this study was to determine the dedicator of cytokinesis 2 (DOCK2), extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun N-terminal kinase-1 (JNK) and Akt signals involved in CXCL13-mediated prostate cancer (PCa) cell invasion and proliferation. MATERIALS AND METHODS: Androgen sensitive (LNCaP), hormone-refractory (PC3) cells and normal cells (RWPE-1) were used to determine CXCL13-mediated PCa cell invasion and proliferation. Immuno blotting, fast activated cell-based (FACE) ELISA, caspase activity, cell invasion and proliferation assays were performed to ascertain some of the signalling events involved in PCa cell proliferation and invasion. RESULTS: Unlike androgen-sensitive LNCaP cells, we report for the first time that the hormone refractory cell line, PC3, expresses DOCK2. CXCL13-mediated LNCaP and PC3 cell invasion was regulated by Akt and ERK1/2 activation in a DOCK2-independent fashion. CXCL13 also promoted LNCaP cell proliferation in a JNK-dependent fashion even in the absence of DOCK2. In contrast, CXCL13 induced PC3 cell proliferation through JNK activation, which required DOCK2. CONCLUSIONS: Our results show CXCL13-mediated PCa cell invasion requires Akt and ERK1/2 activation and suggests a new role for DOCK2 in proliferation of hormone-refractory CXCR5-positive PCa cells. PMID- 21645151 TI - Cholesterol esters as growth regulators of lymphocytic leukaemia cells. AB - OBJECTIVE: Alterations in plasma lipid profile and in intracellular cholesterol homoeostasis have been described in various malignancies; however, significance of these alterations, if any, in cancer biology is not clear. The aim of the present study was to investigate a possible correlation between alterations in cholesterol metabolism and expansion of leukaemia cell numbers. MATERIALS AND METHODS: Lipid profiles in plasma and in primary leukaemia cells isolated from patients with acute or chronic lymphocytic leukaemia (ALL and CLL) were studied. RESULTS AND CONCLUSIONS: Decreased levels of HDL-C were observed in plasma of leukaemic patients, levels of total cholesterol, LDL-C, triglycerides and phospholipids were unchanged or only slightly increased. As compared to normal lymphocytes, freshly isolated leukaemic cells showed increased levels of cholesterol esters and reduction in free cholesterol. Growth stimulation of ALL and CLL cells with phytohemagglutinin led to further increase in levels of cholesterol esters. Conversely, treatment with an inhibitor of cell proliferation such as the mTOR inhibitor, RAD, caused decline in population growth rate of leukaemia cells, which was preceded by sharp reduction in rate of cholesterol esterification. On the other hand, exposure of leukaemic cells to two inhibitors of cholesterol esterification, progesterone and SaH 58-035, caused 60% reduction in their proliferation rate. In addition to demonstrating tight correlation between cell number expansion and cholesterol esterification in leukaemic cells, these results suggest that pathways that control cholesterol esterification might represent a promising targets for novel anticancer strategies. PMID- 21645152 TI - Size-sieved subpopulations of mesenchymal stem cells from intervascular and perivascular equine umbilical cord matrix. AB - OBJECTIVES: Umbilical cord matrix (UCM) has been recently proposed as an alternative source of mesenchymal stem cells (MSCs). The aim of this study was to isolate and characterize presumptive stem cells from intervascular and perivascular equine UCM and to obtain homogeneous subpopulations from both sites. MATERIALS AND METHODS: Umbilical cords were processed for retrieval of MSCs. Unsieved cells from intervascular and perivascular portions were evaluated for cell cycle analysis and for immunophenotyping by flow cytometry. Cells from each site were separated into larger and smaller sieved populations using multi-dishes with 8-MUm pore transwell inserts. Each cell population was characterized in terms of renewal capability, specific marker expression and differentiation potential. Cryopreservation was performed on sieved cells only. RESULTS: Cells from both areas expressed MSC and pluripotential specific markers and were able to differentiate into mesodermic and ectodermic lineages. The sieving procedure yielded two relatively homogeneous subpopulations with comparable characteristics. Surprisingly, after sieving, large intervascular and small perivascular cells were the most rapidly replicating cells [20.53 and 19.49 cell population doublings (PD) after 31 days respectively] and also showed higher fibroblast colony forming unit frequency. Unsieved cell populations were used as controls, and showed PD of 9.42(intervascular cells) and 8.54 (perivascular cells) after 31 days. CONCLUSIONS: Here, cells from UCM represented an intermediate stage between pluripotent embryonic and adult stem cells. Size sieving can be used to isolate more rapidly proliferating cell populations. PMID- 21645153 TI - Two novel aspirin analogues show selective cytotoxicity in primary chronic lymphocytic leukaemia cells that is associated with dual inhibition of Rel A and COX-2. AB - OBJECTIVES: Non-steroidal anti-inflammatory drugs have been shown to induce apoptosis in primary B-cell chronic lymphocytic leukaemia (CLL) cells, but the molecular mechanisms that underpin this observation have not been fully elucidated. Here, we have analysed the effect two novel aspirin analogues, 2 hydroxy benzoate zinc (2HBZ) and 4-hydroxy benzoate zinc (4HBZ), on primary CLL samples. MATERIALS AND METHODS: Cytotoxic effects of 2HBZ and 4HBZ were analysed in primary CLL cells derived from 52 patients, and normal B- and T-lymphocytes. Mechanisms of action of these agents were also elucidated. RESULTS: Both analogues induced apoptosis in a dose-dependent and time-dependent manner. Apoptosis was associated with activation of caspase-3 that could be partially abrogated by the caspase-9 inhibitor (Z-LEHD.fmk). Importantly, both agents demonstrated preferential cytotoxicity in CLL cells when compared to normal B- and T-lymphocytes. In terms of their molecular mechanisms of action, 4HBZ and 2HBZ inhibited COX-2 transcription and protein expression and this was associated with upstream inhibition of transcription factor Rel A. Co-culture of CLL cells with CD40 ligand-expressing mouse fibroblasts significantly increased COX-2 expression and inhibited spontaneous apoptosis. Importantly, the most potent analogue, 4HBZ, overcame pro-survival effects of the co-culture system and significantly repressed COX-2. Finally, elevated COX-2 expression was associated with poor prognostic subsets and increased sensitivity to 4HBZ. CONCLUSIONS: Our results demonstrate therapeutic potential of 4HBZ and are consistent with a mechanism involving suppression of Rel A nuclear translocation and inhibition of COX-2 transcription. PMID- 21645155 TI - Context is everything: the role of auto-ethnography, reflexivity and self critique in establishing the credibility of qualitative research findings. PMID- 21645154 TI - Serine palmitoyltransferase inhibitor myriocin induces growth inhibition of B16F10 melanoma cells through G(2) /M phase arrest. AB - OBJECTIVES: Melanoma is the most aggressive form of skin cancer, and it resists chemotherapy. Candidate drugs for effective anti-cancer treatment have been sought from natural resources. Here, we have investigated anti-proliferative activity of myriocin, serine palmitoyltransferase inhibitor, in the de novo sphingolipid pathway, and its mechanism in B16F10 melanoma cells. MATERIAL AND METHODS: We assessed cell population growth by measuring cell numbers, DNA synthesis, cell cycle progression, and expression of cell cycle regulatory proteins. Ceramide, sphingomyelin, sphingosine and sphingosine-1-phosphate levels were analysed by HPLC. RESULTS: Myriocin inhibited proliferation of melanoma cells and induced cell cycle arrest in the G(2) /M phase. Expressions of cdc25C, cyclin B1 and cdc2 were decreased in the cells after exposure to myriocin, while expression of p53 and p21(waf1/cip1) was increased. Levels of ceramide, sphingomyelin, sphingosine and sphingosine-1-phosphate in myriocin-treated cells after 24 h were reduced by approximately 86%, 57%, 75% and 38%, respectively, compared to levels in control cells. CONCLUSIONS: Our results suggest that inhibition of sphingolipid synthesis by myriocin in melanoma cells may inhibit expression of cdc25C or activate expression of p53 and p21(waf1/cip1) , followed by inhibition of cyclin B1 and cdc2, resulting in G(2) /M arrest of the cell cycle and cell population growth inhibition. Thus, modulation of sphingolipid metabolism by myriocin may be a potential target of mechanism-based therapy for this type of skin cancer. PMID- 21645156 TI - 'I am going to be a dad': experiences and expectations of adolescent and young adult expectant fathers. AB - AIM: To explore the experiences of prospective adolescent fathers regarding their impending fatherhood. BACKGROUND: Several studies indicate adolescent fatherhood is associated with multiple risk factors. Despite this, it is well documented that these young males have a potentially vital role in the lives of their child/ren and partners. Adolescent pregnancy has often been viewed with a particular emphasis on the mother and relatively little is known about the experiences and expectations of young males facing imminent fatherhood. DESIGN: Narrative methods were used to collect qualitative data. METHOD: Narratives were elicited through in-depth interviews with seven adolescent expectant fathers aged 16-22 years. RESULTS: Impending fatherhood presented these young men with mixed emotions and many challenges. The pregnancies were all unplanned and though participants were all willing to face the responsibilities associated with fatherhood, they also reported feeling ill-prepared for the challenges that lay ahead. Impending fatherhood had caused the young men to reflect on the quality of fathering they had received themselves. The young men were hoping to provide their own infant with a better quality of fathering than they had experienced. CONCLUSIONS: Nurses and midwives are ideally placed to provide support to young men facing impending fatherhood to better prepare them to meet the demands of their forthcoming role. RELEVANCE TO CLINICAL PRACTICE: Prenatal classes should include specific sessions for prospective fathers and provide opportunities to assist young men to discuss their thoughts and concerns about impending fatherhood. It could also be useful to encourage young expectant fathers to engage in discussions with their own fathers and grandfathers. For those young men who do not have effective relationships with their own fathers, it could be useful to organise mentoring with experienced mature men who have successfully engaged in the fatherhood role. PMID- 21645157 TI - Jaatha: a fast composite-likelihood approach to estimate demographic parameters. AB - While information about a species' demography is interesting in its own right, it is an absolute necessity for certain types of population genetic analyses. The most widely used methods to infer a species' demographic history do not take intralocus recombination or recent divergence into account, and some methods take several weeks to converge. Here, we present Jaatha, a new composite-likelihood method that does incorporate recent divergence and is also applicable when intralocus recombination rates are high. This new method estimates four demographic parameters. The accuracy of Jaatha is comparable to that of other currently available methods, although it is superior under certain conditions, especially when divergence is very recent. As a proof of concept, we apply this new method to estimate demographic parameters for two closely related wild tomato species, Solanum chilense and S. peruvianum. Our results indicate that these species likely diverged 1.44.N generations ago, where N is the effective population size of S. chilense, and that some introgression between these species continued after the divergence process initiated. Furthermore, S. peruvianum likely experienced a population expansion following speciation. PMID- 21645158 TI - Variation in repeat length and heteroplasmy of the mitochondrial DNA control region along a core-edge gradient in the eastern spadefoot toad (Pelobates syriacus). AB - Peripheral populations are those situated at the distribution margins of a species and are often subjected to more extreme abiotic and biotic conditions than those at the core. Here, we hypothesized that shorter repeat length and fewer heteroplasmic mitochondrial DNA (mtDNA) copies, which are associated with more efficient mitochondrial function, may be related to improved survival under extreme environmental conditions. We sampled eastern spadefoot toads (mostly as tadpoles) from 43 rain pools distributed along a 300-km gradient from core to edge of the species' distribution. We show that mean pool tandem repeat length and heteroplasmy increase from edge to core, even after controlling for body size. We evaluate several alternative hypotheses and propose the Fisher hypothesis as the most likely explanation. However, additional sequential sampling and experimental studies are required to determine whether selection under extreme conditions, or alternative mechanisms, could account for the gradient in heteroplasmy and repeat length in the mtDNA control region. PMID- 21645159 TI - Reconstructing demographic events from population genetic data: the introduction of bumblebees to New Zealand. AB - Four British bumblebee species (Bombus terrestris, Bombus hortorum, Bombus ruderatus and Bombus subterraneus) became established in New Zealand following their introduction at the turn of the last century. Of these, two remain common in the United Kingdom (B. terrestris and B. hortorum), whilst two (B. ruderatus and B. subterraneus) have undergone marked declines, the latter being declared extinct in 2000. The presence of these bumblebees in New Zealand provides an unique system in which four related species have been isolated from their source population for over 100 years, providing a rare opportunity to examine the impacts of an initial bottleneck and introduction to a novel environment on their population genetics. We used microsatellite markers to compare modern populations of B. terrestris, B. hortorum and B. ruderatus in the United Kingdom and New Zealand and to compare museum specimens of British B. subterraneus with the current New Zealand population. We used approximate Bayesian computation to estimate demographic parameters of the introduction history, notably to estimate the number of founders involved in the initial introduction. Species-specific patterns derived from genetic analysis were consistent with the predictions based on the presumed history of these populations; demographic events have left a marked genetic signature on all four species. Approximate Bayesian analyses suggest that the New Zealand population of B. subterraneus may have been founded by as few as two individuals, giving rise to low genetic diversity and marked genetic divergence from the (now extinct) UK population. PMID- 21645160 TI - Fine-scale population structure, inbreeding risk and avoidance in a wild insect population. AB - The ecological and evolutionary importance of fine-scale genetic structure within populations is increasingly appreciated. However, available data are largely restricted to wild vertebrates and eusocial insects. In addition, there is the expectation that most insects tend to have such large- and high-density populations and are so mobile that they are unlikely to face inbreeding risks through fine-scale population structuring. This has made the growing body of evidence for inbreeding avoidance in insects and its implication in mating systems evolution somewhat enigmatic. We present a 4-year study of a natural population of field crickets. Using detailed video monitoring combined with genotyping, we track the movement of all adults within the population and investigate genetic structure at a fine scale. We find some evidence for relatives being found in closer proximity, both across generations and within a single breeding season. Whilst incestuous matings are not avoided, population inbreeding is low, suggesting that mating is close to random and the limited fine scale structure does not create significant inbreeding risk. Hence, there is little evidence for selective pressures associated with the evolution of inbreeding avoidance mechanisms in a closely related species. PMID- 21645161 TI - Spatial structure and the effects of host and soil environments on communities of ectomycorrhizal fungi in wooded savannas and rain forests of Continental Africa and Madagascar. AB - Mycorrhizal fungi play a key role in mineral nutrition of terrestrial plants, but the factors affecting natural distribution, diversity and community composition of particularly tropical fungi remain poorly understood. This study addresses shifts in community structure and species frequency of ectomycorrhizal (EcM) fungi in relation to host taxa, soil depth and spatial structure in four contrasting African ecosystems. We used the rDNA and plastid trnL intron sequence analysis for identification of fungi and host plants, respectively. By partitioning out spatial autocorrelation in plant and fungal distribution, we suggest that African EcM fungal communities are little structured by soil horizon and host at the plant species and family levels. These findings contrast with patterns of vegetation in these forests and EcM fungal communities in other tropical and temperate ecosystems. The low level of host preference indirectly supports an earlier hypothesis that pioneer Phyllanthaceae may facilitate the establishment of late successional Fabaceae and potentially other EcM host trees by providing compatible fungal inoculum in deforested and naturally disturbed ecosystems of tropical Africa. PMID- 21645163 TI - From the editor: is immune contraception in the future? PMID- 21645162 TI - Mechanism of PrP-amyloid formation in mice without transmissible spongiform encephalopathy. AB - Gerstmann-Straussler-Scheinker (GSS) P102L disease is a familial form of a transmissible spongiform encephalopathy (TSE) that can present with or without vacuolation of neuropil. Inefficient disease transmission into 101LL transgenic mice was previously observed from GSS P102L without vacuolation. However, several aged, healthy mice had large plaques composed of abnormal prion protein (PrP(d)). Here we perform the ultrastructural characterization of such plaques and compare them with PrP(d) aggregates found in TSE caused by an infectious mechanism. PrP(d) plaques in 101LL mice varied in maturity, with some being composed of deposits without visible amyloid fibrils. PrP(d) was present on cell membranes in the vicinity of all types of plaques. In contrast to the unicentric plaques seen in infectious murine scrapie, the plaques seen in the current model were multicentric and were initiated by protofibrillar forms of PrP(d) situated on oligodendroglia, astrocytes and neuritic cell membranes. We speculate that the initial conversion process leading to plaque formation begins with membrane-bound PrP(C) but that subsequent fibrillization does not require membrane attachment. We also observed that the membrane alterations consistently seen in murine scrapie and other infectious TSEs were not present in 101LL mice with plaques, suggesting differences in the pathogenesis of these conditions. PMID- 21645164 TI - Contraceptive vaccines: success, status, and future perspective. PMID- 21645165 TI - Current overview of the management of urogenital atrophy in women with breast cancer. AB - Systemic treatments for women with breast cancer frequently induce urogenital symptoms that can negatively impact a women's quality of life. Urogenital atrophy is frequently undiagnosed and untreated, particularly in breast cancer survivors. Symptoms of urogenital atrophy can usually be relieved with vaginal estrogen preparations, but risk of recurrence and safety is undefined in women with a history of breast cancer. Treatment with nonhormonal modalities including vaginal moisturizers or lubricants and lifestyle modification are the first lines of management. Low-dose vaginal 17 beta-estradiol (vaginal estradiol tablets 10 MUg or vaginal estradiol ring) can be considered for the treatment of symptomatic urogenital atrophy in women with a history of breast cancer after appropriate disclosure to the patient. While effective in treating the urogenital symptoms, the safety of such therapy remains uncertain. The decision to offer vaginal estrogen therapy needs to be individualized and should be made jointly with the oncologist. PMID- 21645166 TI - Fine tuning of the Van Nuys prognostic index (VNPI) 2003 by integrating the genomic grade index (GGI): new tools for ductal carcinoma in situ (DCIS). AB - Ductal carcinoma in situ (DCIS) is considered a heterogeneous premalignant condition of the breast with a certain probability for progressing to malignancy. There is no standard of care. The updated Van Nuys Prognostic Index (VNPI) 2003 is a clinical tool in treatment decision making. This study assessed the prognostic value of the VNPI after integration of proliferative biomarkers (GGI and Ki-67). DCIS samples were divided into three VNPI subgroups (low risk [score 4-6], intermediate risk [score 7-9], high risk [score 10-12]) based on nuclear grade +/- necrosis, tumor size, margin width, and age. Nuclear grade was substituted by the genomic grade index (GGI) to generate the VNPI-GGI and combined with the Ki-67 to generate the VNPI-Ki67. Disease-free survival was calculated by Kaplan-Meier survival plots with log-rank significance. Multiple regression analysis was carried out using Cox proportional hazard regression analysis. A total of 88 cases (median age 54 years) with representative tissue were identified out of 168 DCIS patients. Median follow-up was more than 5 years. Ten patients developed an ipsilateral recurrence of whom nine were invasive: six patients were classified in the VNPI subgroup 2 and three patients in the VNPI subgroup 3. One non-invasive recurrence (DCIS) was classified in the VNPI subgroup III. A statistical association was observed between a high VNPI score and a higher risk of recurrence (HR = 7.72 [95% CI 1.01-58.91], p = 0.049). Ki-67 did not improve the prognostic value of VNPI (HR = 6.5, [95% CI 0.80-53.33], p = 0.08). In contrast, the VNPI-GGI could identify more accurately high-risk DCIS patients with early relapses within 5 years (HR = 18.14 [95% CI 1.75-188], p = 0.015). GGI incorporated into the VNPI improved its prognostic value for DCIS, especially for identifying early relapses. This method should be validated and incorporated in future prospective clinical DCIS trials. PMID- 21645167 TI - Pure primary osteosarcoma of the breast. PMID- 21645168 TI - Primary pulmonary synovial sarcoma presenting as a breast mass. PMID- 21645169 TI - A novel technique for localization of sentinel lymph node in breast cancer patients based on computed tomographic lymphography. PMID- 21645170 TI - Developmental breast asymmetry. AB - Developmental breast asymmetry (DBA) can affect psychosocial well-being in the young female. Correction of breast asymmetry may present a reconstructive challenge, especially in tuberous breasts. Fifty-two cases of DBA treated between January 2002 and January 2006 were reviewed. Preoperative clinical assessment of the specific anatomical deformity, subsequent surgical treatment modalities, esthetic outcome, and patient's satisfaction were evaluated. Surgical modalities used in our series include augmentation mammaplasty with or without tissue expansion, parenchymal scoring, nipple areola complex reduction, glanduloplasty techniques, mastopexy and reduction mammaplasty. The mean age of DBA presentation was 21 years; 69% (36/52) patients had tuberous breasts, of which 67% (24/36) were unilateral and 33% (12/36) were bilateral deformities. Patients with tuberous breast deformity presented consistently under the age of 25 years. Esthetic outcome was rated "good" in 75% (39/52), and symmetry rated as "good" in 58% (30/52) by professional evaluation. Surgical treatment is tailored to the affected esthetic units of the individual breast. In our experience, symmetry is the hardest parameter to achieve, particularly in tuberous breasts. Operative treatment is of great value to the psychosocial well-being of the patient. A conceptual approach in the assessment and treatment of DBA is emphasized by this series. PMID- 21645171 TI - Neglected and emerging diseases in sub-Saharan Africa. PMID- 21645172 TI - Diaphragm and peripheral muscle thickness on ultrasound: intra-rater reliability and variability of a methodology using non-standard recumbent positions. AB - BACKGROUND AND OBJECTIVE: Reliable measurement of diaphragm and peripheral muscle thickness, using diagnostic ultrasound, has only been validated in the erect posture. However, in many clinical populations, including critically ill patients, the erect posture presents logistic difficulties. This study aimed to validate ultrasound measurement of diaphragm and peripheral muscle thickness in the recumbent position. METHODS: An observational methodology of repeated but blind ultrasound and anthropometric measurements was applied, to assess inta rater reliability. Thirteen healthy volunteers (aged 20-73years) participated. A pneumotachograph was used to target lung volume, as diaphragm thickness was measured from ultrasound at end-expiration, and both 25% and 50% of inspiratory capacity, while semi-recumbent. The thicknesses of the mid-upper arm, mid-forearm and mid-thigh musculature were also measured bilaterally while supine. RESULTS: Diaphragm thickness could be reliably measured at end-expiration (intra-class correlation coefficient (ICC)=0.990, 95% confidence interval: 0.918-0.998), 25% of inspiratory capacity (ICC=0.959 (0.870-0.988)) and 50% of inspiratory capacity (ICC=0.994 (0.980-0.998)). Peripheral muscle thickness measurements were also reliable (ICC=0.998-1.0). Supine anthropometric measurements of limb segment lengths and girths were highly reproducible. CONCLUSIONS: This ultrasound technique has good reliability in recumbent positions, making it useful for application to clinical populations when the erect posture is not practical. PMID- 21645173 TI - Rituximab monotherapy with eight weekly infusions for relapsed or refractory patients with indolent B cell non-Hodgkin lymphoma mostly pretreated with rituximab: a multicenter phase II study. AB - Information regarding rituximab monotherapy with eight weekly infusions for relapsed or refractory indolent B cell non-Hodgkin lymphoma (B-NHL), in particular for patients pretreated with rituximab, is limited. To evaluate the efficacy and safety of eight doses of rituximab monotherapy, 52 patients with relapsed or refractory indolent B-NHL were enrolled in the present study. Forty of 45 eligible patients (89%) had follicular lymphoma and 24 (53%) were at intermediate or high risk group according to the Follicular Lymphoma International Prognostic Index. The median number of prior chemotherapy regimens was 1 (range 1-7). At the median follow-up of 12.2 months, the overall response rate (ORR), complete response rate (%CR), and median progression-free survival (PFS) were 69% (95% confidence interval [CI] 53%-82%), 47% (95% CI 32%-62%), and 15.6 months (95% CI 10.6- months), respectively. In the 33 patients pretreated with rituximab, the ORR, %CR, and median PFS were inferior compared with values for the 12 patients who had not received rituximab previously (64%vs 83% for ORR; 39%vs 67% for %CR; and 13.8 vs 17.5 months for median PFS, respectively). All mild-to-moderate infusion-related toxicities were reversible. Grade 3/4 non hematologic adverse events occurred in six of the 52 patients. Two patients developed Grade 4 late-onset neutropenia and a decrease (>50%) in serum immunoglobulin was observed in six patients. In conclusion, rituximab monotherapy with eight weekly infusions is effective in relapsed patients with indolent B NHL, with acceptable toxicities, including in patients pretreated with rituximab; however, careful monitoring is recommended for infections associated with late onset neutropenia and hypogammaglobulinemia. (University Hospital Medical Information Network no. UMIN000002974.) PMID- 21645174 TI - Motor features and response to oral levodopa in patients with Parkinson's disease under continuous dopaminergic infusion or deep brain stimulation. AB - BACKGROUND: Subthalamic nucleus deep brain stimulation (STN DBS) and continuous dopaminergic infusions (jejunal levodopa or subcutaneous apomorphine) are indicated in complicated Parkinson's disease (PD), although it remains unsettled how they compare to each other. METHODS: We investigated the daytime motor condition in patients with advanced PD under monotherapy with jejunal levodopa, subcutaneous apomorphine, or STN DBS and also measured the motor changes produced by an additional standard morning dose of levodopa. Motor performance was assessed with the UPDRS-III, hand taps, the AIMS dyskinesia score and patients' diaries. Outcome measures were time to best motor 'on' after start of morning treatment, daytime variability of motor condition, motor scores. RESULTS: The time to 'on' was longest in the jejunal levodopa group. DBS and jejunal levodopa treatments produced stable motor conditions without appreciable 'off' episodes. Continuous apomorphine infusion was associated with the worst motor scores (UPDRS III and taps) and the most frequent off-states. Jejunal levodopa infusion was associated with the highest AIMS scores. Addition of a levodopa dose produced shortening of time to 'on' and a transient motor improvement in the jejunal levodopa group without increase in dyskinesias; in the DBS and apomorphine groups, there was an increase in dyskinesias without changes in UPDRS-III or taps. CONCLUSIONS: STN DBS provided adequate trade-off between motor improvement and dyskinesia control, although dyskinesias could be elicited by adding oral levodopa. Jejunal levodopa infusion produced adequate motor improvement with slow time to 'on' and moderate dyskinesias. Apomorphine infusion produced insufficient motor control and negligible dyskinesias. PMID- 21645175 TI - Cerebrotendinous xanthomatosis in Spain: clinical, prognostic, and genetic survey. AB - BACKGROUND AND PURPOSE: Cerebrotendinous xanthomatosis (CTX) is a rare autosomal recessive disorder caused by mutations in the CYP27A1 gene resulting in sterol-27 hydroxylase deficiency. Current information about CTX is based mainly on case reports, with only few large series reported. Although perceived as a potentially treatable condition, efficacy of chenodeoxycholic acid plus statin therapy remains unclear. To perform a nationwide survey of confirmed cases, with a thorough analysis of genotype-phenotype data and prognostic factors. METHODS: Retrospective review of the clinical and epidemiological aspects and mutations of all the patients diagnosed since 1992 in the main reference centers for genetic testing of CTX in Spain. RESULTS: Twenty-five patients from 19 families were identified. An average delay of 19 years was observed between symptom onset and clinical diagnosis. Two main clinical subgroups were recognizable: a classic form (cerebellar and other supratentorial symptoms) and a spinal form (chronic myelopathy). Cholestanol levels did not correlate with clinical presentation, severity or response to therapy. Despite treatment, five patients died during follow-up, one to 4 years after diagnosis. Thirteen different mutations were identified, with a higher frequency of p.R395C in Northwestern Spain and p.R405W in Southern Spain. None of the mutations could be associated with a particular clinical feature combination or prognosis. CONCLUSIONS: This is the first nationwide extensive series of CTX reported in Spain. The higher number of cases in some areas suggests a possible founder effect. Spinal forms had a less severe prognosis. A delayed diagnosis could contribute to the lack of significant response to treatment. PMID- 21645176 TI - Periventricular and deep white matter leukoaraiosis have a closer association with cerebral microbleeds than age. AB - BACKGROUND: Taking an advantage of the high sensitivity of 3D T2*-weighted gradient-recalled-echo (GRE) imaging to cerebral microbleeds, we investigated the relationship between cerebral microbleeds and leukoaraiosis. METHODS: Participants aged 40 years or more have been evaluated for the presence of cerebral microbleeds using 3D T2*-GRE sequence since 2006. The severity of periventricular hyperintensity (PVH) and deep white matter hyperintensity (DWMH) on fluid attenuated inversion recovery images was assessed using Fazekas rating scales. Multivariate logistic regression analyses were conducted after adjustment for stroke subtype, age, PVH, DWMH, hypertension, dementia, and use of platelet aggregation inhibitors. Additionally, we examined the association between cerebral microbleeds and other covariates using a Pearson's correlation analysis. RESULTS: Amongst 389 patients, 67 patients had a single microbleed and 93 had multiple microbleeds. The prevalence of microbleeds was 83% amongst 53 patients with intracerebral hemorrhage (ICH), 49% amongst 173 with infarction, and 20% amongst 163 without any type of stroke. In the multivariate analyses, the odds ratio (95% CIs) of microbleed detection was 10.1, (4.12-24.8) for ICH, 2.33 (1.12 4.85) for atherosclerotic infarction, 1.66 (1.10-2.48) for PVH, and 1.49 (1.02 2.19) for DWMH. In the Pearson's correlation analysis, cerebral microbleeds were closely related to PVH (Pearson's correlation coefficient; 0.48) and DWMH (0.37), compared with age (0.16). CONCLUSIONS: High-grade PVH, high-grade DWMH, ICH, and atherosclerotic infarction were significantly independent predictors for cerebral microbleeds. In addition, we found that the grades of PVH and DWMH have a closer association with the number of cerebral microbleeds than age. PMID- 21645177 TI - Low risk of vascular events following urgent treatment of transient ischaemic attack: the Aarhus TIA study. AB - BACKGROUND AND PURPOSE: Patients with TIA have a high short-time risk of stroke and an increased long-term risk of ischaemic vascular events compared with the general population. Urgent intervention may reduce short-time stroke risk, but little is known about the effect beyond 3 months. We examined 1-year outcome and risk factor management in patients with TIA after urgent intervention. METHODS: All patients with TIA referred to Aarhus University Hospital 1 March 2007-28 February 2008 were seen by an acute TIA team (ATT), integrating outpatient care and stroke unit facilities. Preventive treatment was initiated immediately, including fast-track surgery for carotid stenosis. Follow-up including nurse conducted health counseling was carried out after 7, 90, and 365 days. RESULTS: A total of 306 patients were included. Stroke, myocardial infarction, or vascular death occurred in 5.2% during 1 year of follow-up. The cumulated stroke rate was 1.6%, 2.0%, and 4.4% after 7, 90, and 365 days, respectively, compared to expected 4.5% [relative risk (RR) 0.36, 95% CI 0.13-0.98] and 7.5% (RR 0.26, 95% CI 0.11-0.63) after 7 and 90 days using ABCD(2) criteria. Recurrent TIA occurred in 10.2% (n = 32). Secondary prevention targets were attained in 47.6% after 1 year. Carotid surgery was performed in 8.1%; median time to operation was 11 days after contact with the ATT. CONCLUSION: Urgent intervention after TIA by an ATT covering outpatient and stroke unit facilities combined with nurse-conducted health counseling is associated with a low 1-year risk of new vascular events and may improve risk factor control. PMID- 21645178 TI - Pharmacokinetics, pharmacodynamics and safety of recombinant canine FVIIa in a study dosing one haemophilia A and one haemostatically normal dog. AB - Recombinant human FVIIa (rhFVIIa) corrects the coagulopathy in hemophilia A and B as well as FVII deficiency. This is also the case in dogs until canine anti-human FVIIa antibodies develop (~2 weeks). Recombinant canine factor VIIa (rcFVIIa), successfully over-expressed by gene transfer in haemophilia dogs, has provided long-term haemostasis (>2 years). However, pharmacokinetics (PK), pharmacodynamics (PD) and safety of rcFVIIa after pharmacological administration have not been reported. We therefore wanted to explore the safety, PK and PD of rcFVIIa in dogs. A pilot study was set up to evaluate the safety as well as PK and PD of rcFVIIa after a single intravenous dose of 270 MUg kg(-1) to one HA and one haemostatically normal dog and to directly compare rcFVIIa with rhFVIIa in these two dogs. Single doses of rcFVIIa and rhFVIIa were well tolerated. No adverse events were observed. Pharmacokinetic characteristics including half-life (FVIIa activity: 1.2-1.8 h; FVIIa antigen 2.8-3.7 h) and clearance were comparable for rcFVIIa and rhFVIIa. Kaolin-activated thromboelastography approached normal in the HA dog with the improvement being most pronounced after rcFVIIa. This study provided the first evidence that administering rcFVIIa intravenously is feasible, safe, well tolerated and efficacious in correcting the haemophilic coagulopathy in canine HA and that rcFVIIa exhibits pharmacokinetic characteristics comparable to rhFVIIa in haemophilic and haemostatically competent dogs. This strengthens the hypothesis that rcFVIIa can be administered to dogs to mimic the administration of rhFVIIa to humans. PMID- 21645179 TI - Assessment and management of pain in haemophilia patients. AB - Haemophilia patients experience acute pain during joint bleeds and chronic pain from haemophilic arthropathy. More than 50% of haemophilia patients have painful joints that cause disability and impair quality of life. Unfortunately, only a few clinical studies have investigated the non-pharmacological or pharmacological treatments for pain or the adverse effects of pain on the health and quality of life of children and adults with haemophilia. There are no detailed algorithms or guidelines for pain management in haemophilia patients, and treatment is largely empirical. Therefore, a standardized approach to the management of pain in haemophilia patients is needed. This approach should include a close relationship between pain specialists and the staffs at haemophilia treatment centres; validated instruments specific to haemophilia for assessing pain, quality of life and disability; and stepwise algorithms/protocols for treatment of chronic vs. acute pain and prophylactic vs early treatment. A pain treatment protocol should include a definition of the problem of pain and best practices for physicians. A call to action is needed to standardize treatment approaches to pain and to develop algorithms/protocols for the management of pain in haemophilia patients. This review will highlight the prevalence and devastating impact of pain in haemophilia patients, currently available treatment options and identify the unmet needs for pain management. PMID- 21645180 TI - Molecular diagnosis of haemophilia A at Centro Hospitalar de Coimbra in Portugal: study of 103 families--15 new mutations. AB - Haemophilia A (HA), the most commonly inherited bleeding disorder, has well known phenotype heterogeneity, influenced by the type of mutation, modulating factors and development of inhibitors. Nowadays, new technologies in association with bioinformatics tools allow a better genotype/phenotype correlation. With the main objective of identifying familial carrier women and to offer prenatal diagnosis, 141 HA patients belonging to 103 families, followed or referred to the Haemophilia Centre of CHC, E.P.E., were studied. Molecular diagnosis strategy was based on HA severity: IVS22 and IVS1 inversions, direct sequencing and MLPA technique. New missense and splicing mutations were further analyzed using molecular modelling. Genotype/phenotype correlation was assessed taking into account the known modulating factors. During this study, mutations were detected in 102/103 families, carrier status was determined in 83 women and 14 prenatal diagnoses were performed. In a total of 46 different mutations identified, 15 have not been reported previously by the HAMSTeRS and HGMD. Genotype/phenotype correlation revealed two cases with a clinical picture less severe than expected by the type of mutation identified. Six patients developed inhibitors: five severe (IVS22, IVS1, large deletion) and one mild (p. Gln2265Lys). The adopted strategy allowed the identification of 99% of the molecular alterations underlying the HA phenotype (98% detection rate for severe and 100% for moderate and mild). Evaluation of genotype-phenotype correlation was complemented with structural protein modelling of newly identified missense mutations, contributing to better understanding of the disease-causing mechanisms and to deepening knowledge on protein structure-function. PMID- 21645181 TI - Inhibition of growth of Trichophyton tonsurans by Lactobacillus reuteri. AB - AIM: The aims of this study were to identify antifungal lactic acid bacteria (LAB) and characterize their activity against the dermatophyte Trichophyton tonsurans. METHODS AND RESULTS: A total of 165 different LAB were isolated and initially screened for anti-Penicillium expansum activity. Five strains, which exhibited strong inhibitory activity, were then tested against the dermatophyte T. tonsurans DSM12285, where they also caused inhibition as observed by large fungal clearing on agar surface. The strongest inhibition was seen with Lactobacillus reuteri R2. When freeze-dried cell-free supernatant powder from this strain was incorporated in culture medium at concentrations >1%, growth of fungal colony was inhibited. Conidia germination was also inhibited under these conditions as determined by microscopy. The anti-T. tonsurans activity of Lact. reuteri R2 was not affected neither by heat treatment nor by proteolytic treatment using pronase E and proteinase K, indicating that the responsible agent(s) were nonproteinaceous in nature. CONCLUSIONS: Lactobacillus reuteri R2 was identified as having strong inhibitory activity against the dermatophyte T. tonsurans DSMZ12285. SIGNIFICANCE AND IMPACT OF THE STUDY: LAB are naturally associated with many foods and are well recognized for their biopreservative properties. The use of these and/or their products may well provide alternative safe approaches for the inhibition of dermatophytic fungi. PMID- 21645182 TI - A real-time qPCR assay to quantify Fusarium graminearum biomass in wheat kernels. AB - AIMS: To develop a real-time PCR assay to quantify Fusarium graminearum biomass in blighted wheat kernels. METHODS AND RESULTS: Primers designed to amplify a gene in the trichothecene biosynthetic cluster (TRI6) were evaluated for sensitivity and specificity. Primer pair Tri6_10F/Tri6_4R specifically and consistently amplified a 245-bp DNA fragment from F. graminearum. A workflow was developed and validated to extract DNA from infested grain. The assay detected as little as 10 MUg of F. graminearum mycelia in 1 g of ground wheat grain with a high correlation between fungal biomass and cycle threshold values (R(2) = 0.9912; = 0.004). In field-inoculated grain, qPCR measurements of biomass correlated closely with deoxynivalenol levels (R = 0.82, P < 0.0001) and two visual techniques to assess grain quality (R = 0.88, P < 0.0001 and R = 0.81, P < 0.0001). CONCLUSIONS: The qPCR assay provided accurate and precise assessments of the amount of F. graminearum biomass in blighted wheat kernels. This method represents a technical advance over other approaches to quantify kernel colonization and real-time PCR detection methodologies for F. graminearum that do not correlate quantification of fungal genomic DNA to biomass. SIGNIFICANCE AND IMPACT OF THE STUDY: Quantifying F. graminearum biomass, especially low levels of growth associated with kernels that are visually asymptomatic, represents a new approach to screen for resistance to kernel infection, an understudied yet potentially important avenue to reduce the impact of head blight. PMID- 21645183 TI - Prevalence and diversity of class 1 integrons and resistance genes in antimicrobial-resistant Escherichia coli originating from beef cattle administered subtherapeutic antimicrobials. AB - AIMS: To characterize class 1 integrons and resistance genes in tetracycline resistant Escherichia coli originating from beef cattle subtherapeutically administered chlortetracycline (A44), chlortetracycline and sulfamethazine (AS700), or no antimicrobials (control). METHODS AND RESULTS: Tetracycline resistant E. coli (control, n = 111; AS700, n = 53; A44, n = 40) were studied. Class 1 integrons, inserted gene cassettes and the presence of other antimicrobial resistance genes, as well as phylogenetic analysis, were performed by PCR, restriction enzyme analysis and sequencing. Susceptibilities to 11 antimicrobials were conducted on all isolates. Prevalence of class 1 integrase was higher (P < 0.001) in isolates from AS700 (33%) and A44 (28%) steers as compared to control (7%). Most integron gene cassettes belonged to the aad or dfr families. Correlations were found between the tet(A) gene and the genetic elements sul1 (r = 0.44), aadA1 (r = 0.61), cat (r = 0.58) and intI1(r = 0.37). Both closely and distantly related isolates harboured integrons with identical gene cassette arrays. CONCLUSIONS: Subtherapeutic administration of chlorotetracycline alone or in combination with sulfamethazine may select for class 1 integrons in bovine tetracycline-resistant E. coli isolates. Vertical spread and horizontal transfer are responsible for the dissemination of a particular type of class 1 integron, but this study could not differentiate if this phenomenon occurred within or outside of the feedlot. Tetracycline-resistant E. coli strains with sul1 and tet(A) genes were more likely to harbour class 1 integrons. SIGNIFICANCE AND IMPACT OF THE STUDY: Subtherapeutic use of chlortetracycline and sulfamethazine may promote the presence of class 1 integrons in tetracycline-resistant E. coli isolated from feedlot cattle. PMID- 21645184 TI - A 24-week, multicentre, open evaluation of the clinical effectiveness of the rivastigmine patch in patients with probable Alzheimer's disease. AB - BACKGROUND: Cholinesterase inhibitors form the mainstay of treatment for persons with mild-to-moderate Alzheimer's disease (AD). The rivastigmine patch may increase compliance and the proportion of patients maintaining an efficacious dose compared with oral cholinesterase inhibitors. OBJECTIVE: To investigate the proportion of patients who reached and maintained the target rivastigmine patch dose compared with the target rivastigmine capsule dose reported in clinical trials. METHODS: This was a multicentre, 24-week, open-label study in persons with probable AD and a Mini-Mental State Examination (MMSE) score of >= 10 and <= 26. The primary outcome was the proportion of patients (ITT population) treated with 9.5 mg/24 h rivastigmine patch for at least 8 weeks at week 24. Secondary outcomes included week 24 MMSE, Alzheimer's Disease Cooperative Study-Clinical Global Impression of Change (ADCS-CGIC), Trail Making Test Part A (TMT-A) and Alzheimer's Disease Cooperative Study-Activities of Daily Living (ADCS-ADL) scores. RESULTS: Overall, 208 participants received treatment and 155 (74.5%) completed the study. Within the ITT population, 147/182 patients (80.8%; 95% CI 75.0-86.5%) were treated for at least 8 weeks with the 9.5 mg/24 h rivastigmine patch; 135/182 patients (74.2%; 95% CI 67.8-80.5%) were treated for at least 8 weeks and completed the study. The most common adverse events were nausea (10.1% of patients), erythema (8.7%), pruritus (8.2%) and vomiting (7.2%). At week 24, patients treated with the rivastigmine patch showed improvements on MMSE, ADCS ADL, ADCS-CGIC and TMT-A scores. Caregivers reported acceptance, preference and satisfaction with the patch. CONCLUSION: Transdermal delivery may allow more patients to reach and maintain therapeutic doses of rivastigmine compared with oral rivastigmine. PMID- 21645185 TI - Mental health advocacy and African and Caribbean men: good practice principles and organizational models for delivery. AB - BACKGROUND: Advocacy has a critical role to play in addressing concerns about access to appropriate mental health care and treatment for African and Caribbean men. AIM: To investigate good practice principles and organizational models for mental health advocacy provision for African and Caribbean men. STUDY DESIGN: The study consisted of: (i) A systematic literature review. Bibliographic and internet searching was undertaken from 1994 to 2006. The inclusion criteria related to mental health, advocacy provision for African and Caribbean men. (ii) Four focus groups with African and Caribbean men to explore needs for and experiences of mental health advocacy. (iii) An investigation into current advocacy provision through a survey of advocacy provision in England, Wales and Northern Ireland. (iv) Twenty-two qualitative stakeholder interviews to investigate the operation of mental health advocacy for this client group. The study was undertaken in partnership with two service user-led organizations and an African Caribbean mental health service. RESULTS: Primary research in this area is scant. Mainstream mental health advocacy services are often poor at providing appropriate services. Services developed by the Black Community and voluntary sector are grounded in different conceptualizations of advocacy and sharper understanding of the needs of African and Caribbean men. The lack of sustainable funding for these organizations is a major barrier to the development of high-quality advocacy for this group, reflecting a lack of understanding about their distinctive role. CONCLUSIONS: The commissioning and provision of mental health advocacy needs to recognize the distinct experiences of African and Caribbean men and develop capacity in the range of organizations to ensure equitable access. PMID- 21645186 TI - Patients' and professionals' experiences and perspectives of obesity in health care settings: a synthesis of current research. AB - BACKGROUND: Obesity-related stigma likely influences how obese people interact with health-care professionals and access health care. AIM: To undertake a synthesis of studies examining the views and experiences of both obese people in relation to their health-care provision and health-care professionals in providing care to obese patients. SEARCH STRATEGY: A systematic search of key electronic databases relating to professional or patient experiences of, or perspectives on, obesity was performed in 2008 and updated in 2010. Reference lists of article bibliographies were searched, along with hand searches of relevant journals. INCLUSION CRITERIA: Studies were screened against explicit inclusion criteria and published between 1990 and 2010. Findings were examined and organized thematically. DATA EXTRACTION: Data were extracted focusing on obesity, stigma and access to health-care services. All included studies were subject to critical appraisal to assess the quality of the research. FINDINGS: Thirty studies were identified. All the studies reported obesity impacting on health-care interactions. Key themes identified were experiences of stigma and feelings of powerlessness, treatment avoidance, psycho-emotional functioning, professional attitudes, confidence and training, variations in health contact time and finally, differences in treatment options and preventative measures. CONCLUSION: Obesity is a stigmatized condition that impacts negatively on the relationship between patients and health-care providers. Given the increasing prevalence of obesity and the range of therapeutic options available, further work is necessary to understand how the presence of obesity affects health-care interactions and decision making. PMID- 21645187 TI - 'Managing patient involvement': provider perspectives on diabetes decision making. AB - BACKGROUND: Most studies of shared decision-making focus on acute treatment or screening decision-making encounters, yet a significant proportion of primary care is concerned with managing patients with chronic disease. AIM: To investigate provider perspectives on the role of patient involvement in chronic disease decision-making. DESIGN: A qualitative, grounded theory study of patient involvement in diabetes care planning. SETTING AND PARTICIPANTS: Interviews were conducted with 29 providers (19 general practitioners, eight allied health providers, and two endocrinologists) who participated in diabetes care planning. RESULTS: Providers described a conflict between their responsibilities to deliver evidence-based diabetes care and to respect patients' rights to make decisions. While all were concerned with providing best possible diabetes care, they differed in the emphasis they placed on 'treating to target' or practicing 'personalized care'. Those preferring to 'treat to target' were more assertive, while 'personalized care' meant being more accepting of the patient's priorities. Providers sought to manage patient involvement in decision-making according to their objectives. 'Treating to target' meant involving patients where necessary to tailor care to their needs and abilities, but limiting patient involvement in decisions about the overall agenda. 'Personalized care' meant involving patients to tailor care to patient preference. DISCUSSION AND CONCLUSIONS: Respecting a patient's autonomy and delivering high-quality diabetes care are important to providers. At times it may not be possible to do both, so a careful balance is required. Involving patients in decision-making may be a means to this end, rather than an end in itself. PMID- 21645188 TI - Blueprint for a deliberative public forum on biobanking policy: were theoretical principles achievable in practice? AB - BACKGROUND: Public deliberation is recommended for obtaining citizen input to policy development when policies involve contested ethical dimensions, diverse perspectives on how to trade-off competing public interests and low public awareness of these perspectives. Several norms have been proposed for the design of deliberative methods. Evidence is scarce regarding whether such norms are achievable in practice. PURPOSE: This paper refers to principles of deliberative democracy theory to describe a deliberative public forum on biobanking. Practical challenges and contextual facilitators of achieving deliberative ideals are discussed, along with factors that influenced use of the forum output in policy development. METHOD: The forum ran for 4 days over two weekends in Perth, Western Australia. Key methodological features were socio-demographic stratification to randomly recruit a mini-public of citizens for discursive representation, provision of information inclusive of diverse perspectives and framed for difference, provision of a fair way for reasoning and collective decision making and adoption of processes to achieve publicity, accountability and independence from undue institutional influence. RESULTS: Most design principles were achieved in practice, with the fundamental exception of representativeness. Factors influencing these outcomes, and the use of deliberated outputs to develop policy, included institutional characteristics, the design involvement of deliberative experts and quality of the outputs when compared to other consultation methods. CONCLUSIONS: Public deliberations can achieve design ideals and influence (ethics based) public health policy. The representation of 'hard to reach' citizens and their views needs further consideration, particularly as this relates to the procedural legitimacy of ethical analyses and the just inclusion of deliberative citizen advice within the broader policy-making process. PMID- 21645189 TI - Stages of change and health-related quality of life among employees of an institution. AB - BACKGROUND: Transtheoretical Model of change has been used successfully in promoting behaviour change. OBJECTIVE: To examine the relationships between health-related quality of life (HRQoL) scores with the stages of change of adequate physical activity and fruit and vegetables intake. DESIGN: This was a cross-sectional study conducted among employees of the main campus and Engineering campus of Universiti Sains Malaysia (USM) during October 2009 and March 2010. MAIN VARIABLES STUDIED: Data on physical activity and fruit and vegetable intake was collected using the WHO STEPS instrument for chronic disease risk factors surveillance. The Short Form-12 health survey (SF-12) was used to gather information on participants' HRQoL. The current stages of change are measured using the measures developed by the Pro-Change Behaviour Systems Incorporation. STATISTICAL ANALYSIS: One way ANOVA and its non-parametric equivalent Kruskal-Wallis were used to compare the differences between SF-12 scores with the stages of change. RESULTS: A total of 144 employees were included in this analysis. A large proportion of the participants reported inadequate fruits and vegetable intake (92.3%) and physical activity (84.6%). Mean physical and mental component scores of SF-12 were 50.39 (SD = 7.69) and 49.73 (SD = 8.64) respectively. Overall, there was no statistical significant difference in the SF 12 domains scores with regards to the stages of change for both the risk factors. CONCLUSIONS: There were some evidence of positive relationship between stages of change of physical activity and fruit and vegetable intake with SF-12 scores. Further studies need to be conducted to confirm this association. PMID- 21645190 TI - Patients' decision making to accept or decline an implantable cardioverter defibrillator for primary prevention of sudden cardiac death. AB - BACKGROUND: Patients are offered implantable defibrillators (ICDs) for the prevention of sudden cardiac death (SCD). However, patients' decision-making process (DMP) of whether or not to accept an ICD has not been explored. We asked patients about their decision making when offered an ICD. DESIGN/SETTING: A grounded theory methodology was employed. Patients were recruited from three ICD centres. Those who received an ICD underwent interviews the first month after implant. Declining patients had interviews at their convenience. In-depth analysis of transcripts was completed. Identified themes were placed along process pathways in a DMP model and tested. FINDINGS: Forty-four patients consented to participate (25% women). Thirty-four accepted an ICD and 10 (23%) declined. Ages ranged from 26 to 87 (mean = 65; SD = 12.5). Participants were retired (65%), had ischaemic heart disease (64%) and some post-secondary education (52%). The DMP was triggered when patient's risk for SCD was communicated. The physician's recommendation and a new awareness SCD risk were motivators to accept the ICD. Patient's decision-making approaches fell along a continuum, from active and engaged to passive and indifferent. Patient's approaches were influenced most by the following: (i) trust; (ii) social influences and (iii) health state. CONCLUSIONS: Health-care providers need to recognize the DMP pathways in which ICD candidacy and SCD risk are understood. The factors that influence a patient's decision warrant discussion pre-implant. It is imperative that patients comprehend the meaning of ICD candidacy to make an informed decision. Participants did not recall alternatives to receiving ICD therapy. PMID- 21645191 TI - Exosome secretion: molecular mechanisms and roles in immune responses. AB - Exosomes are small membrane vesicles, secreted by most cell types from multivesicular endosomes, and thought to play important roles in intercellular communications. Initially described in 1983, as specifically secreted by reticulocytes, exosomes became of interest for immunologists in 1996, when they were proposed to play a role in antigen presentation. More recently, the finding that exosomes carry genetic materials, mRNA and miRNA, has been a major breakthrough in the field, unveiling their capacity to vehicle genetic messages. It is now clear that not only immune cells but probably all cell types are able to secrete exosomes: their range of possible functions expands well beyond immunology to neurobiology, stem cell and tumor biology, and their use in clinical applications as biomarkers or as therapeutic tools is an extensive area of research. Despite intensive efforts to understand their functions, two issues remain to be solved in the future: (i) what are the physiological function(s) of exosomes in vivo and (ii) what are the relative contributions of exosomes and of other secreted membrane vesicles in these proposed functions? Here, we will focus on the current ideas on exosomes and immune responses, but also on their mechanisms of secretion and the use of this knowledge to elucidate the latter issue. PMID- 21645192 TI - Biochemical characterization of APPL endosomes: the role of annexin A2 in APPL membrane recruitment. AB - APPL endosomes are a recently identified subpopulation of early endosomes characterized by the presence of two homologous Rab5 effector proteins APPL1 and APPL2. They exhibit only limited colocalization with EEA1, another Rab5 effector and a marker of the canonical early endosomes. Although APPL endosomes appear to play important roles in cargo trafficking and signal transduction, their protein composition and biochemical properties remain largely unknown. Here we employed membrane fractionation methods to characterize APPL endosomes biochemically. We demonstrate that they represent heterogeneous membrane structures which can be discriminated from the canonical EEA1-positive early endosomes by their partly different physical properties and a distinct migration pattern in the continuous density gradients. In search for other potential markers of APPL endosomes we identified Annexin A2 as an interacting partner of both APPL1 and APPL2. Annexin A2 is a Ca(2+) and phosphatidylinositol 4,5-bisphosphate binding protein, previously implicated in several endocytic steps. We show that Annexin A2 co fractionates and colocalizes with APPL endosomes. Moreover, silencing of its expression causes solubilization of APPL2 from endosomes. Although Annexin A2 is not an exclusive marker of APPL endosomes, our data suggest that it has an important function in membrane recruitment of APPL proteins, acting in parallel to Rab5. PMID- 21645193 TI - Ice age climate, evolutionary constraints and diversity patterns of European dung beetles. AB - Current climate and Pleistocene climatic changes are both known to be associated with geographical patterns of diversity. We assess their associations with the European Scarabaeinae dung beetles, a group with high dispersal ability and well known adaptations to warm environments. By assessing spatial stationarity in climate variability since the last glacial maximum (LGM), we find that current scarab richness is related to the location of their limits of thermal tolerance during the LGM. These limits mark a strong change in their current species richness-environment relationships. Furthermore, northern scarab assemblages are nested and composed of a phylogenetically clustered subset of large-range sized generalist species, whereas southern ones are diverse and variable in composition. Our results show that species responses to current climate are limited by the evolution of assemblages that occupied relatively climatically stable areas during the Pleistocene, and by post-glacial dispersal in those that were strongly affected by glaciations. PMID- 21645194 TI - Synergies between climate anomalies and hydrological modifications facilitate estuarine biotic invasions. AB - Environmental perturbation, climate change and international commerce are important drivers for biological invasions. Climate anomalies can further increase levels of habitat disturbance and act synergistically to elevate invasion risk. Herein, we use a historical data set from the upper San Francisco Estuary to provide the first empirical evidence for facilitation of invasions by climate extremes. Invasive zooplankton species did not become established in this estuary until the 1970s when increasing propagule pressure from Asia coincided with extended drought periods. Hydrological management exacerbated the effects of post-1960 droughts and reduced freshwater inflow even further, increasing drought severity and allowing unusually extreme salinity intrusions. Native zooplankton experienced unprecedented conditions of high salinity and intensified benthic grazing, and life history attributes of invasive zooplankton were advantageous enough during droughts to outcompete native species and colonise the system. Extreme climatic events can therefore act synergistically with environmental perturbation to facilitate the establishment of invasive species. PMID- 21645195 TI - Increase of body weight during the first year of intensive insulin treatment in type 2 diabetes: systematic review and meta-analysis. AB - AIM: This systematic review and meta-analysis was to evaluate the body weight increase and its clinical correlates, through direct weighted regressions, as well as the effect of different insulin regimens and insulin analogues, through meta-analysis. METHODS: Appropriate methodology according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement was adhered to. Forty-six randomized studies, published as full papers, reporting the effect of insulin treatment on change in body weight were identified, and used to identify predictors of weight change; studies were included in a meta-analysis to study the effect of different insulin regimens or insulin analogues on weight change. RESULTS: Intensity of treatment [aim of study (fasting blood glucose, mg/dl), dose of insulin, final HbA1c, change of HbA1c and frequency of hypoglycaemia] was significantly associated with body weight increase, with small differences between basal versus twice-a-day and prandial regimen. At meta analysis, body weight increase was lower with basal regimen than with twice-a-day regimen and than with a prandial regimen. Within all regimens, body weight increase was lower with detemir than with NPH, with no difference between glargine and NPH; only two studies directly compared detemir and glargine, indicating lower weight gain with the former insulin. Within twice-a-day regimens and within prandial regimens, comparison was between newer analogues and older drugs, with no significant difference in body weight increase. CONCLUSION: Body weight increase during the first year of insulin treatment is associated with the intensity of treatment; body weight increase also depends on the insulin regimen applied. PMID- 21645196 TI - Acute systemic inflammation induced by influenza A (H1N1) vaccination causes a deterioration in endothelial function in HIV-infected patients. AB - OBJECTIVES: Vaccination of HIV-infected patients against the influenza A/H1N1 subtype was proposed as a mandatory precautionary measure during the 2009 pandemic. The immediate cardiovascular effects of the novel vaccine have been largely unexplored. We investigated the impact of vaccination on indices of endothelial function in a cohort of HIV-infected patients. METHODS: We included 24 HIV-infected patients in a study with a randomized, sham procedure-controlled design. A monovalent, adjuvanted vaccine against influenza A/H1N1 was used in the vaccine arm (n=16); patients in the control group (n=8) were subjected to a sham procedure. Endothelial function, as assessed by flow-mediated dilatation (FMD), and inflammatory markers were assessed prior to and 8 and 48 h post vaccination. RESULTS: FMD deteriorated following vaccination (baseline, 6.5 +/- 1.1%; 8 h, 1.1 +/- 1.5%; 48 h, 2.0 +/- 1.4%; P=0.04). The white blood cell count increased at 8 h and remained elevated at 48 h. Soluble intercellular adhesion molecule-1 levels decreased after vaccination; the maximum decrease was noted at 48 h. Conversely, the sham procedure did not induce changes in endothelial function or inflammatory markers, apart from a reduction in the white blood cell count at 48 h. CONCLUSIONS: Acute systemic inflammation induced by vaccination against the influenza A/H1N1 virus resulted in a deterioration in endothelial function in HIV infected patients, and this effect was sustained for at least 48 h. Our findings may have important implications in view of the high cardiovascular risk that HIV infection carries. The effect of the novel vaccine on endothelial function should be weighed against the immunological protection that it confers. PMID- 21645197 TI - The efferent segment in continent cutaneous urinary diversion: a comprehensive review of the literature. AB - OBJECTIVE: To critically assess the biophysical properties and current status of outlet formation in heterotopic intestinal urinary diversion. As despite three decades of clinical experience with continent cutaneous urinary diversion through bowel segments, no consensus has been reached for the optimal efferent segment although its function largely determines patient satisfaction. METHODS: A comprehensive Medline literature search using the Medical Subject Headings database (search terms: continent urinary diversion followed by either efferent segment, nipple, Mitrofanoff, Yang-Monti, Benchekroun, tapered ileum, intussuscepted ileum, Kock pouch, T-valve, or Ghonheim) was conducted to identify all full-length original articles addressing the various principles and techniques of outlet formation as well as their outcomes and complications. Examined series were published in English between 1966 and 2010. All studies were systematically evaluated using a checklist (study design, number of patients, etc.) and rated according to the Oxford Centre for Evidence-Based Medicine Levels of Evidence (LoE). RESULTS: While there was a continuous flow of publications over the last three decades, the vast majority of studies were retrospective case series with numerous confounding factors and poorly defined, non-standardized outcomes (LoE, 3). Only a few investigations compare different efferent segments (LoE, 2a). No randomized studies exist. The major biophysical principles are based on the use of flap, nipple, and hydraulic valves. Vermiform appendix, intussuscepted ileal nipple, and the Yang-Monti tube are the most popular techniques and have well-established data on outcomes, complications, and failure rates. Artificial sphincter systems and tissue engineering have provided disappointing results thus far. Most reconstructive strategies are subject to a process of on-going improvement. CONCLUSIONS: The continuous quest for optimization has not led to a single universally applicable efferent segment in continent cutaneous urinary diversion. While all techniques have their unique set of advantages and disadvantages, they will always remain a compromise. Success depends on selecting the optimal strategy for individual patients. A major change in principles in the near future is unlikely. PMID- 21645198 TI - A randomized, blinded, controlled trial of the antiemetic effect of ondansetron on dexmedetomidine-induced emesis in cats. AB - OBJECTIVE: To determine the effect of ondansetron on the incidence of vomiting in cats pre-medicated with dexmedetomidine and buprenorphine. STUDY DESIGN: Randomized, blinded, controlled trial. ANIMALS: Eighty-nine female domestic shorthair cats, aged 3-60 months (median, 12 months) and weighing 1.2-5.1 kg. METHODS: Each cat received dexmedetomidine (40 MUg kg(-1)) plus buprenorphine (20 MUg kg(-1)), intramuscularly as pre-anesthetic medication. Cats were assigned to three treatment groups: ondansetron (0.22 mg kg(-1), intramuscular [IM]), either 30 minutes before the pre-anesthetic medication (ONDA group, n = 31) or with the pre-anesthetic medication (OPM group, n = 30) mixed with the pre-anesthetic medications in the same syringe, or not to receive the antiemetic (control group, n = 28). Emesis was recorded as an all-or-none response. The number of episodes of emesis and the time until onset of the first emetic episode were recorded for each cat. Clinical signs of nausea were recorded whenever they occurred, and a numerical rating scale was used to quantify these signs. Data were analyzed using Kruskal-Wallis and Chi-square test; a Bonferroni correction was made for six comparisons; thus, the two-sided p for significance was 0.05/6 = 0.008. RESULTS: There was a significant reduction in the number of cats vomiting, in the episodes of vomiting/cat, the time elapsed between the premedication and the first vomiting and the severity of nausea in the OPM group compared to the ONDA and control groups. CONCLUSIONS AND CLINICAL RELEVANCE: In cats, the administration of ondansetron (0.22 mg kg(-1)) ameliorates and reduced the severity of dexmedetomidine-induced nausea and vomiting only when it was administered in association with this drug. PMID- 21645199 TI - Antinociceptive activity of pre- versus post-operative intra-articular bupivacaine in goats undergoing stifle arthrotomy. AB - OBJECTIVE: To evaluate the peri-operative analgesic efficacy of intra-articular bupivacaine administered before or after stifle arthrotomy. STUDY DESIGN: Prospective, randomized, blind, placebo-controlled experimental trial. ANIMALS: Thirty-nine healthy goats. METHODS: The goats were allocated randomly to one of three intra-articular treatment groups: group PRE (bupivacaine before and saline after surgery), group POST (saline before and bupivacaine after surgery) and group CON (saline before and after surgery). Anaesthesia was maintained with a constant end-tidal sevoflurane of 2.5%. Intra-operatively heart rate (HR), respiratory rate and mean arterial blood pressure (MAP) after critical surgical events (CSE) were recorded and compared with pre-incision values. Propofol requirements to maintain surgical anaesthesia were recorded. Flunixin was administered for 5 days. Post-operative pain assessment at 20 minutes, 2 hours, 4 hours after recovery and on day 2 and 3 included a multidimensional pain score (MPS), a lameness score and mechanical nociceptive threshold (MNT) testing. Rescue analgesia consisted of systemic opioids. Data were analysed using Kruskal Wallis, Mann-Whitney, Friedman or chi-square tests as appropriate. RESULTS: Intra operatively, group PRE had lower HR and MAP at several CSEs than groups POST/CON and required less propofol [0 mg kg(-1) (0-0 mg kg(-1))] than group POST/CON [0.3 mg kg(-1) (0-0.6 mg kg(-1))]. Post-operatively, group POST had significantly higher peri-articular MNTs than groups PRE and CON up to 4 hours after recovery. No treatment effect was detected for MPS, lameness scores and rescue analgesic consumption at any time point. CONCLUSIONS AND CLINICAL RELEVANCE: Pre-operative intra-articular bupivacaine provided notable intra-operative analgesia in goats undergoing stifle arthrotomy but did not reduce post-operative pain. Post operative intra-articular bupivacaine provided a short lasting reduction of peri articular hyperalgesia without affecting the requirements for systemic analgesia. Multimodal perioperative pain therapy is recommended to provide adequate analgesia for stifle arthrotomy in goats. PMID- 21645200 TI - Enhancing the expression of starch synthase class IV results in increased levels of both transitory and long-term storage starch. AB - Starch is an important renewable raw material with an increasing number of applications. Several attempts have been made to obtain plants that produce modified versions of starch or higher starch yield. Most of the approaches designed to increase the levels of starch have focused on the increment of the amount of ADP-glucose or ATP available for starch biosynthesis. In this work, we show that the overexpression of starch synthase class IV (SSIV) increases the levels of starch accumulated in the leaves of Arabidopsis by 30%-40%. In addition, SSIV-overexpressing lines display a higher rate of growth. The increase in starch content as a consequence of enhanced SSIV expression is also observed in long-term storage starch organs such as potato tubers. Overexpression of SSIV in potato leads to increased tuber starch content on a dry weight basis and to increased yield of starch production in terms of tons of starch/hectare. These results identify SSIV as one of the regulatory steps involved in the control of the amount of starch accumulated in plastids. PMID- 21645201 TI - Discovery of polymorphisms in starch-related genes in rice germplasm by amplification of pooled DNA and deeply parallel sequencing. AB - High-throughput sequencing of pooled DNA was applied to polymorphism discovery in candidate genes involved in starch synthesis. This approach employed semi- to long-range PCR (LR-PCR) followed by next-generation sequencing technology. A total of 17 rice starch synthesis genes encoding seven classes of enzymes, including ADP-glucose pyrophosphorylase (AGPase), granule starch synthase (GBSS), soluble starch synthase (SS), starch branching enzyme (BE), starch debranching enzyme (DBE) and starch phosphorylase (SPHOL) and phosphate translocator (GPT1) from 233 genotypes were PCR amplified using semi- to long-range PCR. The amplification products were equimolarly pooled and sequenced using massively parallel sequencing technology (MPS). By detecting single nucleotide polymorphism (SNP)/Indels in both coding and noncoding areas of the genes, we identified genetic differences and characterized the SNP/Indel variation and distribution patterns among individual starch candidate genes. Approximately, 60.9 million reads were generated, of which 54.8 million (90%) mapped to the reference sequences. The average coverage rate ranged from 12,708 to 38,300 times for SSIIa and SSIIIb, respectively. SNPs and single/multiple-base Indels were analysed in a total assembled length of 116,403 bp. In total, 501 SNPs and 113 Indels were detected across the 17 starch-related loci. The ratio of synonymous to nonsynonymous SNPs (Ka/Ks) test indicated GBSSI and isoamylase 1 (ISA1) as the least diversified (most purified) and conservative genes as the studied populations have been through cycles of selection. This report demonstrates a useful strategy for screening germplasm by MPS to discover variants in a specific target group of genes. PMID- 21645202 TI - Human alpha-mannosidase produced in transgenic tobacco plants is processed in human alpha-mannosidosis cell lines. AB - Deficiency in human lysosomal alpha-mannosidase (MAN2B1) results in alpha mannosidosis, a lysosomal storage disorder; patients present a wide range of neurological, immunological, and skeletal symptoms caused by a multisystemic accumulation of mannose-containing oligosaccharides. Here, we describe the expression of recombinant MAN2B1 both transiently in Nicotiana benthamiana leaves and in the leaves and seeds of stably transformed N. tabacum plants. After purification from tobacco leaves, the recombinant enzyme was found to be N glycosylated and localized in vacuolar compartments. In the fresh leaves of tobacco transformants, MAN2B1 was measured at 10,200 units/kg, and the purified enzyme from these leaves had a specific activity of 32-45 U/mg. Furthermore, tobacco-produced MAN2B1 was biochemically similar to the enzyme purified from human tissues, and it was internalized and processed by alpha-mannosidosis fibroblast cells. These results strongly indicate that plants can be considered a promising expression system for the production of recombinant MAN2B1 for use in enzyme replacement therapy. PMID- 21645203 TI - Overexpression of Brassica juncea wild-type and mutant HMG-CoA synthase 1 in Arabidopsis up-regulates genes in sterol biosynthesis and enhances sterol production and stress tolerance. AB - Brassica juncea 3-hydroxy-3-methylglutaryl-CoA synthase (HMGS) is encoded by four isogenes (BjHMGS1-BjHMGS4). In vitro enzyme assays had indicated that the recombinant BjHMGS1 H188N mutant lacked substrate inhibition by acetoacetyl-CoA (AcAc-CoA) and showed 8-fold decreased enzyme activity. The S359A mutant demonstrated 10-fold higher activity, while the H188N/S359A double mutant displayed a 10-fold increased enzyme activity and lacked inhibition by AcAc-CoA. Here, wild-type and mutant BjHMGS1 were overexpressed in Arabidopsis to examine their effects in planta. The expression of selected genes in isoprenoid biosynthesis, isoprenoid content, seed germination and stress tolerance was analysed in HMGS overexpressors (OEs). Those mRNAs encoding enzymes 3-hydroxy-3 methylglutaryl-CoA reductase (HMGR), sterol methyltransferase 2 (SMT2), delta-24 sterol reductase (DWF1), C-22 sterol desaturase (CYP710A1) and brassinosteroid-6 oxidase 2 (BR6OX2) were up-regulated in HMGS-OEs. The total sterol content in leaves and seedlings of OE-wtBjHMGS1, OE-S359A and OE-H188N/S359A was significantly higher than OE-H188N. HMGS-OE seeds germinated earlier than wild type and vector-transformed controls. HMGS-OEs further displayed reduced hydrogen peroxide (H(2) O(2) )-induced cell death and constitutive expression of salicylic acid (SA)-dependent pathogenesis-related genes (PR1, PR2 and PR5), resulting in an increased resistance to Botrytis cinerea, with OE-S359A showing the highest and OE-H188N the lowest tolerance. These results suggest that overexpression of HMGS up-regulates HMGR, SMT2, DWF1, CYP710A1 and BR6OX2, leading to enhanced sterol content and stress tolerance in Arabidopsis. PMID- 21645204 TI - Management of hepatitis B virus infection in the underprivileged world. PMID- 21645205 TI - To MMP or not to MMP: a role for matrix metalloproteinase 3 in primary sclerosing cholangitis? PMID- 21645206 TI - Treatment of chronic hepatitis B virus infection in resource-constrained settings: expert panel consensus. AB - Most of the estimated 350 million people with chronic hepatitis B virus (HBV) infection live in resource-constrained settings. Up to 25% of those persons will die prematurely of hepatocellular carcinoma (HCC) or cirrhosis. Universal hepatitis B immunization programmes that target infants will have an impact on HBV-related deaths several decades after their introduction. Antiviral agents active against HBV are available; treatment of HBV infection in those who need it has been shown to reduce the risk of HCC and death. It is estimated that 20-30% of persons with HBV infection could benefit from treatment. However, drugs active against HBV are not widely available or utilized in persons infected with HBV. Currently recommended antiviral agents used for treatment of human immunodeficiency virus (HIV) infection do not adequately suppress HBV, which is of great concern for the estimated 10% of the HIV-infected persons in Africa who are co-infected with HBV. Progressive liver disease has been shown to occur in co infected persons whose HBV infection is not suppressed. In view of these concerns, an informal World Health Organization consultation of experts concluded that: chronic HBV is a major public health problem in emerging nations; all HIV infected persons should be screened for HBV infection; HIV/HBV co-infected persons should be treated with therapies active against both viruses and that reduce the risk of resistance; standards for the management of chronic HBV infection should be adapted to resource-constrained settings. In addition, a research agendum was developed focusing on issues related to prevention and treatment of chronic HBV in resource-constrained settings. PMID- 21645207 TI - 15th International Symposium on Cells of the Hepatic Sinusoid, 2010. AB - This is a meeting report of the presentations given at the 15th International Symposium on Cells of the Hepatic Sinusoid, held in 2010. The areas covered include the contributions of the various liver cell populations to liver disease, molecular and cellular targets involved in steatohepatitis, hepatic fibrosis and cancer and regenerative medicine. In addition to a review of the science presented at the meeting, this report provides references to recent literature on the topics covered at the meeting. PMID- 21645208 TI - Use of hepatocyte and stem cells for treatment of post-resectional liver failure: are we there yet? AB - Post-operative liver failure following extensive resections for liver tumours is a rare but significant complication. The only effective treatment is liver transplantation (LT); however, there is a debate about its use given the high mortality compared with the outcomes of LT for chronic liver diseases. Cell therapy has emerged as a possible alternative to LT especially as endogenous hepatocyte proliferation is likely inhibited in the setting of prior chemo/radiotherapy. Both hepatocyte and stem cell transplantations have shown promising results in the experimental setting; however, there are few reports on their clinical application. This review identifies the potential stem cell sources in the body, and highlights the triggering factors that lead to their mobilization and integration in liver regeneration following major liver resections. PMID- 21645209 TI - Impact of tumour differentiation to select patients before liver transplantation for hepatocellular carcinoma. AB - AIM: To generate a new score with improved accuracy compared with Milan criteria to select patients. PATIENTS: The training cohort comprised 373 patients transplanted for hepatocellular carcinoma (HCC) between 1988 and 1998 (cohort 1). An algorithm was derived from the analysis of patient data by the proportional hazard Cox regression model. The area under the receiver operating characteristic (AUROC) was used to determine a cut-off value. The validation cohort comprised 140 patients transplanted between 1999 and 2001 (cohort 2). RESULTS: Multivariate analysis identified three predictors of 5-year tumour-free survival: tumour differentiation (P=0.02), diameter (P<0.0001) and number of nodules (P=0.04). A cut-off value of 4 was derived from the AUROC of the final score. Five-year tumour-free survival was 60.2 +/- 3.1% in patients with as score <4 and 36.4 +/- 4.7% in individuals with a score >=4, P<0.0001. In the validation cohort, 5-year tumour-free survival was 82.8 +/- 3.6% (score <4) and 50.0 +/- 10.7% (score >=4), P=0.0003. In patients with a score <4, there was no significant difference in 5 year tumour-free survival between Milan+ and Milan- patients, either in cohort 1 or 2. Five-year tumour-free survival of Milan- patients was significantly better in individuals with a score <4 compared with those with a score >=4, both in cohort 1 (61.5 +/- 9.1 vs 31.4 +/- 4.6%, P=0.009) and in cohort 2 (P=0.02). CONCLUSION: A novel score taking into account tumour differentiation shows higher accuracy than Milan criteria in predicting 5-year tumour-free survival following liver transplantation for HCC. Prospective studies should validate these findings. PMID- 21645210 TI - XRCC1 genetic polymorphism Arg399Gln and hepatocellular carcinoma risk: a meta analysis. AB - BACKGROUND: Studies investigating the association between X-ray repair cross complementing group 1 (XRCC1) genetic polymorphism Arg399Gln and hepatocellular carcinoma (HCC) risk report conflicting results. The aim of this study was to quantitatively summarize the evidence for such a relationship. METHODS: Two investigators independently searched the Medline, Embase, CNKI and Chinese Biomedicine Database. Summary odds ratios (ORs) and 95% confidence intervals (95% CIs) for XRCC1 polymorphism and HCC were calculated in a fixed-effects model (the Mantel-Haenszel method) and a random-effects model (the DerSimonian and Laird method) when appropriate. The pooled ORs were performed for a codominant model (Gln/Gln vs. Arg/Arg, Arg/Gln vs. Arg/Arg), a dominant model (Gln/Gln+Arg/Gln vs. Arg/Arg) and a recessive model (Gln/Gln vs. Arg/Gln+Arg/Arg). RESULTS: This meta analysis included 11 case-control studies, which included 2208 HCC cases and 3265 controls. Overall, the variant genotypes (Gln/Gln and Arg/Gln) of Arg399Gln were not associated with HCC risk when compared with the wild-type Arg/Arg homozygote (Gln/Gln vs. Arg/Arg, OR=1.01, 95% CI=0.79-1.28; Arg/Gln vs. Arg/Arg, OR=1.09, 95% CI=0.81-1.45). Similarly, no associations were found in the dominant and recessive models (dominant model, OR=1.12, 95% CI=0.85-1.47; recessive model, OR=0.99, 95% CI=0.79-1.25). Limiting the analysis to the studies within Hardy Weinberg equilibrium, the results were persistent and robust. When stratifying for ethnicity, country/region and source of controls, no evidence of a significant association was observed in any subgroup. No publication bias was found in the present study. CONCLUSION: No association is found between the XRCC1 polymorphism Arg399Gln and the risk of HCC. PMID- 21645211 TI - AKR1B10 expression is associated with less aggressive hepatocellular carcinoma: a clinicopathological study of 168 cases. AB - BACKGROUND/AIMS: The detoxification enzyme AKR1B10, a member of the aldo-keto reductase superfamily, is discussed as a new biomarker candidate for hepatocellular carcinoma (HCC). Only rare clinicopathological data on AKRB1B10 in HCC exist. This retrospective study determines the diagnostic and prognostic relevance of AKR1B10 expression in HCC and its relationship to a series of clinicopathological parameters including underlying aetiological factors. METHODS: A series of 168 patients with HCCs treated either by surgical resection (n=92) or liver transplantation (n=76) were investigated after construction of a tissue micro-array. Immunohistochemically confirmed AKR1B10 expression was correlated with clinicopathologically relevant parameters as well as proliferative activity (indicated by Ki-67 immunostaining) and apoptosis (terminal deoxyribonucleotide transferase-mediated dUTP nick-end labelling). RESULTS: AKR1B10 overexpression is significantly associated with lower pT classification (P=0.030) and highly statistically associated with an underlying viral hepatitis (P<0.001) and the presence of cirrhosis (P<0.001). In addition, loss of AKR1B10 expression correlates with increased proliferative activity (Ki 67, P=0.001). Kaplan-Meier survival analysis of the resection group reveals a poorer prognosis in patients with AKR1B10-negative HCCs compared with patients with strongly positive HCCs (P=0.046). CONCLUSIONS: This study confirms and expands data on the expression of AKR1B10 in HCC, suggesting that this enzyme is a valuable novel biomarker candidate for staging of HCC, especially in patients with underlying virus hepatitis or cirrhosis, and may present a new therapeutic target for multimodal therapy concepts. We confirm its prognostic value and conclude that high expression of AKR1B10 reflects a less aggressive tumour behaviour. PMID- 21645212 TI - The change of the quantitative HBsAg level during the natural course of chronic hepatitis B. AB - BACKGROUND: There is insufficient information about HBsAg levels and their correlation with serum hepatitis B virus (HBV) DNA in chronic hepatitis B (CHB). AIMS: We aimed to describe HBsAg levels during various phases of CHB and to investigate the correlation with serum HBV DNA levels. METHODS: A total of 645 treatment-naive Korean CHB patients were included in this retrospective cross sectional study. They were categorized into immune tolerance (IT, n=56), HBeAg positive hepatitis (EPH, n=150), inactive carrier (IC, n=274) and HBeAg-negative hepatitis (ENH, n=165). The baseline HBsAg and HBV DNA levels were measured. RESULTS: The mean HBsAg titres (log IU/ml) differed (P<0.001): IT 4.29, EPH 3.64, IC 2.05 and ENH 3.23. In 645 patients, HBsAg and HBV DNA showed a significant correlation (r=0.693, P<0.001), and this was also observed in the IT, EPH and IC groups (r=0.664, r=0.541, r=0.505, respectively, all P<0.001), but not in the ENH group (r=0.093, P=0.321). Age had a negative correlation with HBsAg (r=-0.451, P<0.001). The cirrhotic patients had a significantly lower HBsAg level than the non-cirrhotic patients (2.41 +/- 1.36 vs. 3.02 +/- 1.21 log IU/ml, P<0.001). CONCLUSIONS: The HBsAg level varied significantly in different phases of CHB and was correlated with HBV DNA during the IT, EPH and IC phases. These findings can provide additional information to understand the natural course and pathogenesis of CHB. PMID- 21645213 TI - Evaluation of normal or minimally elevated alanine transaminase, age and DNA level in predicting liver histological changes in chronic hepatitis B. AB - BACKGROUND: Serum alanine transaminase (ALT), hepatitis B virus (HBV) DNA level and age are used in the evaluation of chronic hepatitis B (CHB). AIM: We designed this study to evaluate liver histology with ALT and its relation with age and HBV DNA. METHODS: During the period of October 2006 to July 2009, 499 CHB patients were included in this study with detectable HBV DNA at PCR. Of these, 181 had normal ALT, 200 had ALT [>(1 * ULN) < (2 ULN)] and 118 had ALT >= 2 ULN and were labelled as Group 1, 2 and 3 respectively. RESULTS: A strong positive correlation was found between ALT and histological activity index (HAI) and fibrosis. However, 29 (52.7%) and five (9.1%) in Group 1 with positive HBeAg status had HAI >=4 and fibrosis >=2 respectively. Among those with HBeAg-negative status, 66 (23.1%) had HAI >4 and 31 (10.8%) had fibrosis >=2. In Group 2, 14 (15.7%) had moderate-to-severe HAI and 19 (21.2%) had fibrosis >=2 when HBeAg was positive, in those with HBeAg negative 34 (30.6%) had moderate-to-severe HAI and 38 (34.2%) had fibrosis >=2. An ALT value of >=58.5 U/l had higher sensitivity than that of 80 U/l in predicting significant histological changes. Further, HAI and fibrosis were significantly greater in the age of >30 years. CONCLUSIONS: We recommend liver biopsy in HBeAg-negative CHB over 30 years of age regardless of ALT level and starting treatment at ALT 1.5 * ULN instead of 2 * ULN. PMID- 21645214 TI - Distinct clinical courses according to presenting phenotypes and their correlations to ATP7B mutations in a large Wilson's disease cohort. AB - INTRODUCTION AND AIMS: Wide phenotypic and genotypic heterogeneities in Wilson's disease (WD) have been reported, hampering the study of their correlations. The goal of this study was to identify the factors related to these diversities. METHODS: Clinical courses and molecular genetic characteristics were analysed in 237 unrelated Korean WD families. The average follow-up period was 8.2 +/- 5.8 years. RESULTS: Presenting phenotypes were classified as H1 (12.2%), H2 (42.4%), N1 (21.6%), N2 (0.4%), NX (0.4%), presymptomatic (22.4%) and other (0.4%), modifying the guidelines by Ferenci and colleagues. Age at presentation was youngest and cirrhosis was rarest in the presymptomatic group. Decompensated cirrhosis was the highest in the H1 group. Favourable outcome was rarest in the N1 group. Forty-seven (11 novel) ATP7B mutations were identified in 85% of the 474 alleles. Multiplex ligation-dependent probe amplification assays in ATP7B and analyses of ATOX1 and COMMD1 genes identified no additional mutations. Yeast complementation assays demonstrated functional perturbation of the seven novel missense mutants. Five major mutations, p.Arg778Leu, p.Ala874Val, p.Asn1270Ser, p.Lys838SerfsX35 and p.Leu1083Phe, accounted for 63% of the alleles. H1 was more common, age at presentation was younger and N1+N2+NX tended to be less common in patients with nonsense, frame shifting or splicing mutations than in those with missense mutations alone. Patients with both mutations in the transduction (Td) or the ATP hinge domain showed presymptomatic or hepatic manifestations but no neurological manifestation. CONCLUSIONS: The presenting phenotype strongly affects the clinical outcome of WD, and is related to the ATP7B mutation type and location, providing an evidence for genotype-phenotype correlations in WD. PMID- 21645215 TI - Serum chemokine receptor CXCR3 ligands are associated with progression, organ dysfunction and complications of chronic liver diseases. AB - BACKGROUND: Chemokines are chemotactic mediators that are implicated in liver diseases. In viral hepatitis and primary biliary cirrhosis, a predominant chemokine receptor expressed in the liver is CXCR3, suggesting that its specific ligands are important in the progression of chronic liver diseases across different aetiologies. METHODOLOGY/PRINCIPAL FINDINGS: We analysed the serum concentrations of the CXCR3 ligands, CXCL9 (monokine induced by interferon gamma), CXCL10 (interferon-gamma-inducible protein 10) and CXCL11 (interferon inducible T cell alpha chemo-attractant) in healthy controls (n=53), subjects with histologically determined liver fibrosis (n=109) and patients with different stages of cirrhosis (n=153) of various disease aetiologies. Chemokine concentrations were determined by cytometric bead assay or ELISA respectively. Serum concentrations of all three chemokines were significantly increased in patients with chronic liver diseases compared with healthy controls (P<0.001). In the biopsied fibrosis cohort, CXCL9 and CXCL10 were positively associated with the severity of liver fibrosis (histology and serum markers), while CXCL11 was not. In cirrhotic patients, CXCL9 was increased in early Child-Pugh stages, while CXCL11 was elevated only in Child B and C patients and CXCL10 across all stages. Notably, CXCR3 chemokines were also associated with the development of clinical complications of cirrhosis, especially portal hypertension. All chemokines significantly correlated with serum levels of the hepatoprotective cytokines interleukin (IL)-6 and IL-10, suggesting their involvement in a counter regulatory response during the progression of liver disease, shedding new light on their involvement in the pathophysiology of chronic liver diseases. CONCLUSIONS: CXCR3 chemokines are differentially expressed during chronic liver diseases across different disease stages and aetiologies. Their association with portal hypertension and hepatoprotective cytokines implies biological functions beyond immune cell recruitment, thereby provoking new diagnostic and therapeutic concepts. PMID- 21645216 TI - Haemodynamic derangement in human immunodeficiency virus-infected patients with hepatitis C virus-related cirrhosis: the role of bacterial translocation. AB - BACKGROUND AND AIMS: Analysis of the influence of the effects of increased intestinal permeability on haemodynamic alterations in human immunodeficiency virus (HIV)-infected patients with decompensated hepatitis C virus (HCV)-related liver disease. METHODS: Forty HIV/HCV co-infected patients and 40 HCV mono infected patients, 20 of them with compensated cirrhosis and 20 with a previous decompensation, and 20 healthy controls, were studied. Intestinal permeability was determined by serum levels of lipopolysaccharide-binding protein (LBP). Monocyte expression of toll-like receptor 4 (TLR-4), serum levels of interleukin (IL)-6 and soluble receptors of tumour necrosis factor (sTNFRI) were analysed. Cardiac index, systemic vascular resistance (SVR), plasma renin activity (PRA) and aldosterone concentration were also determined in cirrhotic patients. RESULTS: Serum levels of LBP, TLR-4, IL-6 and sTNFRI were significantly higher in HIV-HCV co-infected and HCV mono-infected patients with decompensated cirrhosis compared with those with compensated liver disease. Significantly lower values of SVR and higher values of cardiac index, PRA and aldosterone concentration were observed in patients with decompensated cirrhosis compared with those with compensated liver disease, particularly in those with elevated levels of IL-6. There were no significant differences between HIV/HCV co-infected and HCV mono infected patients. CONCLUSIONS: Higher intestinal permeability and consequent macrophage activation is observed in patients with cirrhosis; this permeability is even higher in those with portal hypertension. Serum values of IL-6 are associated with the characteristic haemodynamic derangement observed in advanced phases of cirrhosis. HIV/HCV co-infected cirrhotic patients present inflammatory and systemic haemodynamic alterations similar to those observed in HCV mono infected patients. PMID- 21645217 TI - 'Cystic' lesion in the porta hepatis after liver transplantation. PMID- 21645218 TI - Role of cannabinoid receptors in alcoholic hepatic injury: steatosis and fibrogenesis are increased in CB2 receptor-deficient mice and decreased in CB1 receptor knockouts. AB - BACKGROUND: Alcohol is a common cause of hepatic liver injury with steatosis and fibrosis. Cannabinoid receptors (CB) modulate steatosis, inflammation and fibrogenesis. To investigate the differences between CB(1) and CB(2) in the hepatic response to chronic alcohol intake, we examined CB knockout mice (CB(1)( /-), CB(2)(-/-)). METHODS: Eight- to 10-week-old CB(1)(-/-), CB(2)(-/-) and wild type mice received 16% ethanol for 35 weeks. Animals receiving water served as controls. We analysed triglyceride and hydroxyproline contents in liver homogenates. mRNA levels of CBs, pro-inflammatory cytokines [tumour necrosis factor (TNF)-alpha, monocyte chemotactic protein (MCP)-1, interleukin (IL)-1beta] and profibrotic factors [alpha-smooth muscle actin (alpha-SMA), procollagen-Ia, platelet-derived growth factor beta receptor (PDGFbeta-R)] were analysed by reverse transcription-polymerase chain reaction (RT-PCR). Histology (hemalaun and eosin, oil-red O, CD3, CD45R, CD45, F4/80, Sirius red) characterized hepatic steatosis, inflammation and fibrosis. Activation of lipogenic pathways, activation and proliferation of hepatic stellate cell (HSC) were assessed by western blot [fatty acid synthase (FAS), sterol regulatory element binding protein 1c (SREBP-1c), alpha-SMA, proliferating cell nuclear antigen (PCNA), cathepsin D]. RESULTS: Hepatic mRNA levels of the respective CBs were increased in wild-type animals and in CB(1)(-/-) mice after ethanol intake. Ethanol intake in CB(2)(-/-) mice induced much higher steatosis (SREBP-1c mediated) and inflammation (B-cell predominant infiltrates) compared with wild-type animals and CB(1)(-/-) mice. HSC activation and collagen production were increased in all groups after forced ethanol intake, being most pronounced in CB(2)(-/-) mice and least pronounced in CB(1)(-/-) mice. DISCUSSION: The fact that CB(2) receptor knockout mice exhibited the most pronounced liver damage after ethanol challenge indicates a protective role of CB(2) receptor expression in chronic ethanol intake. By contrast, in CB(1) knockouts, the effect of ethanol was attenuated, suggesting aggravation of fibrogenesis and SREBP-1c-mediated steatosis via CB(1) receptor expression after ethanol intake. PMID- 21645219 TI - Anti-ulcer agent teprenone inhibits hepatitis C virus replication: potential treatment for hepatitis C. AB - BACKGROUND: Previously we reported that 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, statins, inhibited hepatitis C virus (HCV) RNA replication. Furthermore, recent reports revealed that the statins are associated with a reduced risk of hepatocellular carcinoma and lower portal pressure in patients with cirrhosis. The statins exhibited anti-HCV activity by inhibiting geranylgeranylation of host proteins essential for HCV RNA replication. Geranylgeranyl pyrophosphate (GGPP) is a substrate for geranylgeranyltransferase. Therefore, we examined the potential of geranyl compounds with chemical structures similar to those of GGPP to inhibit HCV RNA replication. METHODS: We tested geranyl compounds [geranylgeraniol, geranylgeranoic acid, vitamin K(2) and teprenone (Selbex)] for their effects on HCV RNA replication using genome-length HCV RNA-replicating cells (the OR6 assay system) and a JFH-1 infection cell culture system. Teprenone is the major component of the anti-ulcer agent, Selbex. We also examined the anti-HCV activities of the geranyl compounds in combination with interferon (IFN)-alpha or statins. RESULTS: Among the geranyl compounds tested, only teprenone exhibited anti-HCV activity at a clinically achievable concentration. However, other anti-ulcer agents tested had no inhibitory effect on HCV RNA replication. The combination of teprenone and IFN-alpha exhibited a strong inhibitory effect on HCV RNA replication. Although teprenone alone did not inhibit geranylgeranylation, surprisingly, statins' inhibitory action against geranylgeranylation was enhanced by cotreatment with teprenone. CONCLUSIONS: The anti-ulcer agent teprenone inhibited HCV RNA replication and enhanced statins' inhibitory action against geranylgeranylation. This newly discovered function of teprenone may improve the treatment of HCV-associated liver diseases as an adjuvant to statins. PMID- 21645220 TI - Increased myoendothelial gap junctions mediate the enhanced response to epoxyeicosatrienoic acid and acetylcholine in mesenteric arterial vessels of cirrhotic rats. AB - BACKGROUND: Cirrhotic portal hypertension is characterized by mesenteric arterial vasodilation and hyporeactivity to vasoconstrictors. AIM: We evaluated the role of epoxyeicosatrienoic acid (EET) and of myoendothelial gap junctions (GJ) in the haemodynamic alterations of experimental cirrhosis. METHODS: Thirty-five control rats and 35 rats with carbon tetrachloride (CCl(4))-induced cirrhosis were studied. Small resistance mesenteric arteries (diameter <350 MUm) were connected to a pressure servo controller in a video-monitored perfusion system. Concentration-response curves to acetylcholine (ACh) were evaluated in mesenteric arteries pre-incubated with indomethacin, N(G)-nitro-L-arginine-methyl-ester and 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one before and after the epoxygenase inhibitor miconazole or 18alpha-glycyrrhetinic acid (18alpha-GA) (GJ inhibitor). EC(50) was calculated. Concentration-response curves to 11,12-EET were also evaluated. mRNA and protein expression of connexins (Cxs) in the mesenteric arteries was evaluated by real-time PCR and immunohistochemistry. RESULTS: The ACh response was increased in cirrhotic rats (EC(50): -6.55+/-0.10 vs. -6.01+/ 0.10 log[M]; P<0.01) and was blunted by miconazole only in cirrhotic animals. 18alpha-GA blunted the response to ACh more in cirrhotic than that in control rats (P<0.05). Concentration-response curves to 11,12-EET showed an increased endothelium-dependent vasodilating response in cirrhotic rats (P<0.05); the BK(Ca) inhibitor Iberiotoxin (25 nM) blocked the response in normal rats but not in cirrhotic rats, while 18alpha-GA blunted the response in cirrhotic rats but not in control rats. An increased mRNA and protein expression of Cx40 and Cx43 in cirrhotic arteries was detected (P<0.05). CONCLUSIONS: The increased nitric oxide/PGI(2)-independent vasodilation of mesenteric arterial circulation in cirrhosis is because of, at least in part, hyperreactivity to 11,12-EET through an increased expression of myoendothelial GJs. PMID- 21645221 TI - S-adenosyl-L-methionine inhibits collagen secretion in hepatic stellate cells via increased ubiquitination. AB - BACKGROUND: Liver fibrosis is the excessive accumulation of extracellular matrix (ECM) components that disrupt normal liver microcirculation and lead to organ injury. Hepatic stellate cells (HSCs), following transdifferentiation, are the central mediators of hepatic fibrosis through increased secretion of ECM components, including type I collagen. AIMS: The mechanism(s) by which the antioxidant S-adenosyl-L-methionine (SAMe) acts to modulate type I collagen secretion in activated HSCs was examined. METHODS: Hepatic stellate cells were culture-activated for 13-15 days and treated with SAMe. Type I collagen, proteasomal activity and resident endoplasmic reticulum (ER) protein [78-kDa glucose-regulated protein (Grp78) and protein disulphide isomerase (PDI)] expression were measured. Nuclear factor-kappaB (NF-kappaB) activity, and its role in SAMe-mediated collagen inhibition, was determined. Type I collagen polyubiquitination was examined. RESULTS: S-adenosyl-L-methionine significantly inhibited type I collagen secretion without significant changes in type I collagen mRNA expression. SAMe also increased NF-kappaB activity, and blocking NF kappaB activity using a dominant-negative IkappaBalpha abolished the SAMe mediated type I collagen secretion. Examination of the post-transcriptional fate of procollagen demonstrated that SAMe treatment led to intracellular type I collagen polyubiquitination accompanied by diminution of proteasomal activity. Expression of Grp78 and PDI (resident ER proteins) were significantly decreased by SAMe treatment. CONCLUSIONS: S-adenosyl-L-methionine inhibits collagen processing leading to increased ubiquitination and decreased secretion. These findings represent a novel mechanism for modulating type I collagen expression in activated HSCs. PMID- 21645222 TI - Abnormal liver chemistry in patients with influenza A H1N1. PMID- 21645223 TI - Response-guided therapy for acute hepatitis C: applying concepts from chronic treatment. PMID- 21645224 TI - Successful immune tolerance induction by FVIII in hemophilia A patients with inhibitor may occur without deletion of FVIII-specific T cells. AB - BACKGROUND: The development of an inhibitor is the major complication facing patients with hemophilia A treated by administration of factor (F) VIII concentrates. Restoration of tolerance to FVIII can be achieved by prolonged administration of FVIII (immune tolerance induction, ITI). Although ITI has been used for more than 30years in patients with hemophilia A and inhibitor, its mechanism of action is still poorly understood. OBJECTIVES: As administration of high doses of antigen can induce the apoptosis of the T cells recognizing the antigen, a potential mechanism of action of ITI may be the deletion of FVIII specific T cells. PATIENTS/METHODS: We studied the CD4+ T-cell response to FVIII in five (one mild, one moderate and three severe) patients successfully desensitized by administration of FVIII and in control subjects. RESULTS: Following repeated stimulation with autologous dendritic cells loaded with FVIII, FVIII-specific T oligoclonal cell lines were expanded from the blood of one of the successfully desensitized patients. The FVIII-specific T cells produced IL-5, IL-13 and IL-2. By contrast, FVIII-specific T-cell lines could not be derived from three patients with mild hemophilia A without inhibitor or from four normal control subjects. CONCLUSIONS: These data represent the first analysis of the cellular mechanisms regulating the induction of tolerance to FVIII. They demonstrate that successful tolerance induction may occur without deletion of FVIII-specific T cells. PMID- 21645225 TI - The thrombomodulin analog Solulin promotes reperfusion and reduces infarct volume in a thrombotic stroke model. AB - BACKGROUND: Currently there is no approved anticoagulant for treating acute stroke. This is largely because of concern for hemorrhagic complications, and suggests a critical need for safer anticoagulants. Solulin is a soluble analog of the endothelial cell receptor thrombomodulin, able to bind free thrombin and convert it to an activator of the anticoagulant, protein C. OBJECTIVE: Solulin was tested for its ability to inhibit middle cerebral artery occlusion (MCAO) induced by photothrombosis, and to restore MCA patency after establishment of stable occlusion. METHODS: Cerebral blood flow (CBF) was monitored by laser Doppler for 1.5 h after occlusion and again 72 h later. RESULTS: Solulin treatment 30 min before thrombosis resulted in an approximately 50% increase in time to form a stable occlusion. When administered 30 or 60 min after MCAO, Solulin significantly improved CBF within 90 min of treatment. In contrast, none of the vehicle-treated mice showed restoration of CBF in the first 90 min and only 17% did so by 72 h. Solulin treatment was associated with a significant reduction in infarct volume, and was well tolerated with no overt hemorrhage observed in any treatment group. Mechanistic studies in mice homozygous for the factor (F)V Leiden mutation, suggest that Solulin's efficacy derives primarily from the anticoagulant activity of the thrombin-Solulin complex and not from direct anti-inflammatory or neuroprotective effects of Solulin or activated protein C. CONCLUSIONS: Our data indicate that Solulin is a safe and effective anticoagulant that is able to antagonize active thrombosis in acute ischemic stroke, and to reduce infarct volume. PMID- 21645227 TI - Vicinal thiols are required for activation of the alphaIIbbeta3 platelet integrin. AB - BACKGROUND: Closely spaced thiols in proteins that interconvert between the dithiol form and disulfide bonds are called vicinal thiols. These thiols provide a mechanism to regulate protein function. We previously found that thiols in both alphaIIb and beta3 of the alphaIIbbeta3 fibrinogen receptor were required for platelet aggregation. METHODS AND RESULTS: Using p-chloromercuribenzene sulfonate (pCMBS) we provide evidence that surface thiols in alphaIIbbeta3 are exposed during platelet activation. Phenylarsine oxide (PAO), a reagent that binds vicinal thiols, inhibits platelet aggregation and labeling of sulfhydryls in both alphaIIb and beta3. For the aggregation and labeling studies, binding of PAO to vicinal thiols was confirmed by reversal of PAO binding with the dithiol reagent 2,3-Dimercapto-1-propanesulfonic acid (DMPS). In contrast, the monothiol beta-mercaptoethanol did not reverse the effects of PAO. Additionally, PAO did not inhibit sulfhydryl labeling of the monothiol protein albumin, confirming the specificity of PAO for vicinal thiols in alphaIIbbeta3. As vicinal thiols represent redox sensitive sites that can be regulated by reducing equivalents from the extracellular or cytoplasmic environment, they are likely to be important in regulating activation of alphaIIbbeta3. Additionally, when the labeled integrin was passed though a lectin column containing wheat germ agglutinin and lentil lectin a substantial amount of non-labeled alphaIIbbeta3 eluted separately from the labeled receptor. This suggests that two populations of integrin exist on platelets that can be distinguished by thiol labeling. CONCLUSION: A vicinal thiol-containing population of alphaIIbbeta3 provides redox sensitive sites for regulation of alphaIIbbeta3. PMID- 21645226 TI - Most factor VIII B domain missense mutations are unlikely to be causative mutations for severe hemophilia A: implications for genotyping. AB - BACKGROUND & OBJECTIVE: The factor VIII (FVIII) B domain shares very little amino acid homology with other known proteins and is not directly necessary for procoagulant activity. Despite this, missense mutations within the B domain have been reported in patients with hemophilia A. Given that the B domain is dispensable for secretion and function of FVIII, we hypothesized that these mutations should not be causative of hemophilia A in these patients. METHODS: Plasmid vectors containing B domain missense mutations that were reported to be associated with moderate/severe hemophilia A (T751S, D826E, V993L, H1047Y, T1353A, N1441K, L1462P, E1579D, A1591S, P1641L and S1669L) were analyzed for their effect on synthesis and secretion compared with FVIII wild-type (WT) following transient transfection into COS-1 and CHO cells in vitro. Further, H1047Y, N1441K and E1579D mutants were expressed in vivo in a hemophilia A mouse model by hydrodynamic tail-vein injection. RESULTS: FVIII activity and antigen levels for all mutants expressed into the conditioned media of COS-1 and CHO cells were similar to FVIII WT. Also, plasma expression of these mutants was similar to FVIII WT in hemophilia A mice. An in vivo tail clip bleeding assay also demonstrated that blood loss from hemophilia A mice expressing FVIII WT, H1047Y, N1441K and E1579D was similar. CONCLUSIONS: We conclude that most missense mutations within the FVIII B domain would be unlikely to lead to severe hemophilia A and that the majority of such missense mutations represent polymorphisms or non-pathologic mutations. PMID- 21645228 TI - ISTH overt disseminated intravascular coagulation score in patients with septic shock: automated immunoturbidimetric soluble fibrin assay vs. D-dimer assay. PMID- 21645229 TI - More on: lupus anticoagulant testing during acute thrombotic events. PMID- 21645230 TI - Is obstetric thromboprophylaxis with low-molecular-weight heparin effective? Yes, if administered properly. PMID- 21645232 TI - Experts' opinion or the serotonin release assay as a gold standard for the diagnosis of heparin-induced thrombocytopenia (HIT)? PMID- 21645233 TI - More on: calibration for the measurement of microparticles: needs, interests, and limitations of calibrated polystyrene beads for flow cytometry-based quantification of biological microparticles. PMID- 21645234 TI - More on: calibration for the measurement of microparticles: value of calibrated polystyrene beads for flow cytometry-based sizing of biological microparticles. PMID- 21645235 TI - Outpatient treatment in patients with acute pulmonary embolism: the Hestia Study. AB - BACKGROUND: Traditionally, patients with pulmonary embolism (PE) are initially treated in the hospital with low molecular weight heparin (LMWH). The results of a few small non-randomized studies suggest that, in selected patients with proven PE, outpatient treatment is potentially feasible and safe. OBJECTIVE: To evaluate the efficacy and safety of outpatient treatment according to predefined criteria in patients with acute PE. PATIENTS AND METHODS: A prospective cohort study of patients with objectively proven acute PE was conducted in 12 hospitals in The Netherlands between 2008 and 2010. Patients with acute PE were triaged with the predefined criteria for eligibility for outpatient treatment, with LMWH (nadroparin) followed by vitamin K antagonists. All patients eligible for outpatient treatment were sent home either immediately or within 24 h after PE was objectively diagnosed. Outpatient treatment was evaluated with respect to recurrent venous thromboembolism (VTE), including PE or deep vein thrombosis (DVT), major hemorrhage and total mortality during 3 months of follow-up. RESULTS: Of 297 included patients, who all completed the follow-up, six (2.0%; 95% confidence interval [CI] 0.8-4.3) had recurrent VTE (five PE [1.7%] and one DVT [0.3%]). Three patients (1.0%, 95% CI 0.2-2.9) died during the 3 months of follow-up, none of fatal PE. Two patients had a major bleeding event, one of which was fatal intracranial bleeding (0.7%, 95% CI 0.08-2.4). CONCLUSION: Patients with PE selected for outpatient treatment with predefined criteria can be treated with anticoagulants on an outpatient basis. (Dutch Trial Register No 1319; http://www.trialregister.nl/trialreg/index.asp). PMID- 21645236 TI - Progranulin contributes to endogenous mechanisms of pain defense after nerve injury in mice. AB - Progranulin haploinsufficiency is associated with frontotemporal dementia in humans. Deficiency of progranulin led to exaggerated inflammation and premature aging in mice. The role of progranulin in adaptations to nerve injury and neuropathic pain are still unknown. Here we found that progranulin is up regulated after injury of the sciatic nerve in the mouse ipsilateral dorsal root ganglia and spinal cord, most prominently in the microglia surrounding injured motor neurons. Progranulin knockdown by continuous intrathecal spinal delivery of small interfering RNA after sciatic nerve injury intensified neuropathic pain like behaviour and delayed the recovery of motor functions. Compared to wild-type mice, progranulin-deficient mice developed more intense nociceptive hypersensitivity after nerve injury. The differences escalated with aging. Knockdown of progranulin reduced the survival of dissociated primary neurons and neurite outgrowth, whereas addition of recombinant progranulin rescued primary dorsal root ganglia neurons from cell death induced by nerve growth factor withdrawal. Thus, up-regulation of progranulin after neuronal injury may reduce neuropathic pain and help motor function recovery, at least in part, by promoting survival of injured neurons and supporting regrowth. A deficiency in this mechanism may increase the risk for injury-associated chronic pain. PMID- 21645237 TI - Sequential use of human-derived medium supplements favours cardiovascular tissue engineering. AB - For clinical application of tissue engineering strategies, the use of animal derived serum in culture medium is not recommended, because it can evoke immune responses in patients. We previously observed that human platelet-lysate (PL) is favourable for cell expansion, but generates weaker tissue as compared to culture in foetal bovine serum (FBS). We investigated if human serum (HS) is a better human supplement to increase tissue strength. Cells were isolated from venous grafts of 10 patients and expanded in media supplemented with PL or HS, to determine proliferation rates and expression of genes related to collagen production and maturation. Zymography was used to assess protease expression. Collagen contraction assays were used as a two-dimensional (2D) model for matrix contraction. As a prove of principle, 3D tissue culture and tensile testing was performed for two patients, to determine tissue strength. Cell proliferation was lower in HS-supplemented medium than in PL medium. The HS cells produced less active matrix metallo-proteinase 2 (MMP2) and showed increased matrix contraction as indicated by gel contraction assays and 3D-tissue culture. Tensile testing showed increased strength for tissues cultured in HS when compared to PL. This effect was more pronounced if cells were sequentially cultured in PL, followed by tissue culture in HS. These data suggest that sequential use of PL and HS as substitutes for FBS in culture medium for cardiovascular tissue engineering results in improved cell proliferation and tissue mechanical properties, as compared to use of PL or HS apart. PMID- 21645238 TI - Role of Slug transcription factor in human mesenchymal stem cells. AB - The pathways that control mesenchymal stem cells (MSCs) differentiation are not well understood, and although some of the involved transcription factors (TFs) have been characterized, the role of others remains unclear. We used human MSCs from tibial plateau (TP) trabecular bone, iliac crest (IC) bone marrow and Wharton's jelly (WJ) umbilical cord demonstrating a variability in their mineral matrix deposition, and in the expression levels of TFs including Runx2, Sox9, Sox5, Sox6, STAT1 and Slug, all involved in the control of osteochondroprogenitors differentiation program. Because we reasoned that the basal expression level of some TFs with crucial role in the control of MSC fate may be correlated with osteogenic potential, we considered the possibility to affect the hMSCs behaviour by using gene silencing approach without exposing cells to induction media. In this study we found that Slug-silenced cells changed in morphology, decreased in their migration ability, increased Sox9 and Sox5 and decreased Sox6 and STAT1 expression. On the contrary, the effect of Slug depletion on Runx2 was influenced by cell type. Interestingly, we demonstrated a direct in vivo regulatory action of Slug by chromatin immunoprecipitation, showing a specific recruitment of this TF in the promoter of Runx2 and Sox9 genes. As a whole, our findings have important potential implication on bone tissue engineering applications, reinforcing the concept that manipulation of specific TF expression levels may elucidate MSC biology and the molecular mechanisms, which promote osteogenic differentiation. PMID- 21645239 TI - Plasma membrane microdomains regulate TACE-dependent TNFR1 shedding in human endothelial cells. AB - Upon stimulation by histamine, human vascular endothelial cells (EC) shed a soluble form of tumour necrosis factor receptor 1 (sTNFR1) that binds up free TNF, dampening the inflammatory response. Shedding occurs through proteolytic cleavage of plasma membrane-expressed TNFR1 catalysed by TNF-alpha converting enzyme (TACE). Surface expressed TNFR1 on EC is largely sequestered into specific plasma membrane microdomains, the lipid rafts/caveolae. The purpose of this study was to determine the role of these domains in TACE-mediated TNFR1 shedding in response to histamine. Human umbilical vein endothelial cells derived EA.hy926 cells respond to histamine via H1 receptors to shed TNFR1. Both depletion of cholesterol by methyl-beta-cyclodextrin and small interfering RNA knockdown of the scaffolding protein caveolin-1 (cav-1), treatments that disrupt caveolae, reduce histamine-induced shedding of membrane-bound TNFR1. Moreover, immunoblotting of discontinuous sucrose gradient fractions show that TACE, such as TNFR1, is present within low-density membrane fractions, concentrated within caveolae, in unstimulated EA.hy926 endothelial cells and co-immunoprecipitates with cav-1. Silencing of cav-1 reduces the levels of both TACE and TNFR1 protein and displaces TACE, from low-density membrane fractions where TNFR1 remains. In summary, we show that endothelial lipid rafts/caveolae co-localize TACE to surface expressed TNFR1, promoting efficient shedding of sTNFR1 in response to histamine. PMID- 21645240 TI - Angiotensin II receptor type 1 blockade decreases CTGF/CCN2-mediated damage and fibrosis in normal and dystrophic skeletal muscles. AB - Connective tissue growth factor (CTGF/CCN-2) is mainly involved in the induction of extracellular matrix (ECM) proteins. The levels of CTGF correlate with the degree and severity of fibrosis in many tissues, including dystrophic skeletal muscle. The CTGF overexpression in tibialis anterior skeletal muscle using an adenoviral vector reproduced many of the features observed in dystrophic muscles including muscle damage and regeneration, fibrotic response and decrease in the skeletal muscle strength. The renin-angiotensin system is involved in the genesis and progression of fibrotic diseases through its main fibrotic components angiotensin-II and its transducer receptor AT-1. The use of AT-1 receptor blockers (ARB) has been shown to decrease fibrosis. In this paper, we show the effect of AT-1 receptor blockade on CTGF-dependent biological activity in skeletal muscle cells as well as the response to CTGF overexpression in normal skeletal muscle. Our results show that in myoblasts ARB decreased CTGF-mediated increase of ECM protein levels, extracellular signal regulated kinases 1/2 (ERK 1/2) phosphorylation and stress fibres formation. In tibialis anterior muscle overexpressing CTGF using an adenovirus, ARB treatment decreased CTGF-mediated increase of ECM molecules, alpha-SMA and ERK-1/2 phosphorylation levels. Quite remarkable, ARB was able to prevent the loss of contractile force of tibialis anterior muscles overexpressing CTGF. Finally, we show that ARB decreased the levels of fibrotic proteins, CTGF and ERK-1/2 phosphorylation augmented in a dystrophic skeletal muscle from mdx mice. We propose that ARB is a novel pharmacological tool that can be used to decrease the fibrosis induced by CTGF in skeletal muscle associated with muscular dystrophies. PMID- 21645241 TI - Management of a permanent tooth after trauma to deciduous predecessor: an evaluation by cone-beam computed tomography. AB - Impaction of permanent teeth represents a clinical challenge with regard to diagnosis, treatment plan, and prognosis. There is a close relationship between deciduous teeth and permanent teeth germ, and any injury in the deciduous dentition may influence the permanent teeth eruption. The extent of the damage caused to the permanent teeth germ depends on the patient age at the time of injury, type of trauma, severity, and direction of the impact. Conventional radiographic images are frequently used for diagnosis; however, recent developments in three-dimensional (3D) imaging systems have enabled dentistry to visualize structural changes effectively, with better contrast and more details, close to the reality. The cone-beam computed tomography (CBCT) has been used in the diagnosis and treatment plan of these impacted teeth. The purpose of the present case report is to describe a successful conservative management of a retained permanent maxillary lateral incisor with delayed root development after a trauma through the deciduous predecessor in a 9 year-old patient. After clinical and radiographic examination, a CBCT examination of the maxilla was requested to complement the diagnosis, providing an accurate 3D position of the retained tooth and its relationship to adjacent structures. The proposed treatment plan was the surgical exposure and orthodontic traction of the retained tooth. The lateral incisor spontaneously erupted after 6 months. Therefore, this case report suggests that permanent teeth with incomplete root formation have a great potential for spontaneous eruption because no tooth malposition or mechanical obstacles are observed. PMID- 21645242 TI - Tooth embedded in tongue following firearm trauma: report of two cases. AB - Injuries caused by projectiles from firearms involve diverse patterns of dentoalveolar trauma due to the different types of wound and extent of tissue damage. This article reports two cases in which tooth fragments were embedded in the tongue following aggression from a firearm projectile in the facial region. Radiographs confirmed the presence of foreign bodies, which were surgically removed under local anesthesia. When dentoalveolar trauma occurs in facial injuries, both hard and soft tissues must be carefully examined to avoid overlooking embedded tooth fragments not located immediately in the soft tissue. PMID- 21645243 TI - Has transplantation lost its luster? AB - Has transplantation, once the innovative darling of medicine, lost its novelty for young surgeons and physicians? "The AJT Report" takes a look at the shortage of trainees in transplant medicine and surgery and possible ways to solve the problem. Also this month, new OPTN/UNOS officers and board members take their posts. PMID- 21645244 TI - Microbial pathophysiology and opportunistic infections: surprises ahead. PMID- 21645245 TI - Evaluating the risk of cancer transmission to optimize organ usage. PMID- 21645246 TI - Feces in our food, viruses in our organs: donor surveillance, organ transplantation and the risk for disease transmission. PMID- 21645247 TI - Creating a safer donor: a quarter century of progress? PMID- 21645248 TI - Miles to go.... PMID- 21645249 TI - Laparoscopic kidney transplantation-novel or novelty? PMID- 21645250 TI - Antibody-mediated rejection--an ounce of prevention is worth a pound of cure. AB - The presence of preformed, donor-specific alloantibodies inpatients undergoing renal transplantation is associated with a high risk of hyperacute and acute antibody-mediated rejection (ABMR), and often limits potential recipients' access to organs from living and deceased donors. Over the last decade, understanding of ABMR has improved markedly and given rise to numerous, diverse strategies for the transplantation of allosensitized recipients. Antibody desensitization programs have been developed to allow renal transplant recipients with a willing but antibody-incompatible living donor to undergo successful transplantation, whereas kidney paired exchange schemes circumvent the antibody incompatibility altogether by finding suitable pairs to donors and recipients. Recognizing the complexity of ABMR and the recent developments that have occurred in this important clinical research field, the Roche Organ Transplantation Research Foundation (ROTRF) organized a symposium during the XXIII Congress of The Transplantation Society in Vancouver, Canada, to discuss current understanding in ABMR and ways to prevent it. This Meeting Report summarizes the presentations of the symposium, which addressed key areas that included the interactions between alloantibodies and the complement system in mediating graft injury, technological advancements for assessing antibody-mediated immune responses to HLA antigens, and the potential benefits and challenges of desensitization and kidney paired donation schemes. PMID- 21645251 TI - Donor-transmitted malignancies in organ transplantation: assessment of clinical risk. AB - The continuing organ shortage requires evaluation of all potential donors, including those with malignant disease. In the United States, no organized approach to assessment of risk of donor tumor transmission exists, and organs from such donors are often discarded. The ad hoc Disease Transmission Advisory Committee (DTAC) of the Organ Procurement and Transplantation Network/United Network for Organ Sharing (OPTN/UNOS) formed an ad hoc Malignancy Subcommittee to advise on this subject. The Subcommittee reviewed the largely anecdotal literature and held discussions to generate a framework to approach risk evaluation in this circumstance. Six levels of risk developed by consensus. Suggested approach to donor utilization is given for each category, recognizing the primacy of individual clinical judgment and often emergent clinical circumstances. Categories are populated with specific tumors based on available data, including active or historical cancer. Benign tumors are considered in relation to risk of malignant transformation. Specific attention is paid to potential use of kidneys harboring small solitary renal cell carcinomas, and to patients with central nervous system tumors. This resource document is tailored to clinical practice in the United States and should aid clinical decision making in the difficult circumstance of an organ donor with potential or proven neoplasia. PMID- 21645252 TI - Characterization of the withdrawal phase in a porcine donation after the cardiac death model. AB - Transplantation of donation after cardiac death (DCD) livers has higher rates of organ failure and complications, specifically ischemic biliary injuries. Reported large animal DCD models all employ active means to halt circulation, contrary to human DCD protocol. We report a DCD porcine model in which the animal passively progresses to cardiac death, thereby more closely mimicking human DCD scenario. Sixteen Yorkshire pigs (10 females, 6 males, 30-45 kg) had a mean time of 26:19 min +/- 14:14 from withdrawal of ventilatory support (WVS) to circulatory arrest and 44:38 min +/- 16:37 from WVS to electrical standstill. Cessation of hepatic flow (HF) occurred well before electrical standstill (22:15 min +/- 10:09), previously not described in human or animal DCD. Histologically comparing livers from our DCD model demonstrated a dramatic increase in hepatocyte vacuolization, disorganization of endoplasmic reticulum, formation of mitochondrial inclusions and apoptosis compared with control specimens. Subtle changes were also evident in biliary epithelial cells (BEC). This results in severe cellular changes before reperfusion. Early histologic evidence suggests that there is severe hepatocyte and biliary cell disruption in our DCD model. Further research using this model may provide a deeper understanding of the pathophysiology of the DCD liver. PMID- 21645253 TI - Estimated risk of human immunodeficiency virus and hepatitis C virus infection among potential organ donors from 17 organ procurement organizations in the United States. AB - To prevent unintentional transmission of bloodborne pathogens through organ transplantation, organ procurement organizations (OPOs) screen potential donors by serologic testing to identify human immunodeficiency virus (HIV) and hepatitis C virus (HCV) infection. Newly acquired infection, however, may be undetectable by serologic testing. Our objective was to estimate the incidence of undetected infection among potential organ donors and to assess the significance of risk reductions conferred by nucleic acid testing (NAT) versus serology alone. We calculated prevalence of HIV and HCV-stratified by OPO risk designation-in 13,667 potential organ donors managed by 17 OPOs from 1/1/2004 to 7/1/2008. We calculated incidence of undetected infection using the incidence-window period approach. The prevalence of HIV was 0.10% for normal risk potential donors and 0.50% for high risk potential donors; HCV prevalence was 3.45% and 18.20%, respectively. For HIV, the estimated incidence of undetected infection by serologic screening was 1 in 50,000 for normal risk potential donors and 1 in 11,000 for high risk potential donors; for HCV, undetected incidence by serologic screening was 1 in 5000 and 1 in 1000, respectively. Projected estimates of undetected infection with NAT screening versus serology alone suggest that NAT screening could significantly reduce the rate of undetected HCV for all donor risk strata. PMID- 21645254 TI - Transmission of human immunodeficiency virus and hepatitis C virus from an organ donor to four transplant recipients. AB - In 2007, a previously uninfected kidney transplant recipient tested positive for human immunodeficiency virus type 1 (HIV) and hepatitis C virus (HCV) infection. Clinical information of the organ donor and the recipients was collected by medical record review. Sera from recipients and donor were tested for serologic and nucleic acid-based markers of HIV and HCV infection, and isolates were compared for genetic relatedness. Routine donor serologic screening for HIV and HCV infection was negative; the donor's only known risk factor for HIV was having sex with another man. Four organs (two kidneys, liver and heart) were transplanted to four recipients. Nucleic acid testing (NAT) of donor sera and posttransplant sera from all recipients were positive for HIV and HCV. HIV nucleotide sequences were indistinguishable between the donor and four recipients, and HCV subgenomic sequences clustered closely together. Two patients subsequently died and the transplanted organs failed in the other two patients. This is the first recognized cotransmission of HIV and HCV from an organ donor to transplant recipients. Routine posttransplant HIV and HCV serological testing and NAT of recipients of organs from donors with suspected risk factors should be considered as routine practice. PMID- 21645255 TI - Mechanisms of donor-specific tolerance in recipients of haploidentical combined bone marrow/kidney transplantation. AB - We recently reported long-term organ allograft survival without ongoing immunosuppression in four of five patients receiving combined kidney and bone marrow transplantation from haploidentical donors following nonmyeloablative conditioning. In vitro assays up to 18 months revealed donor-specific unresponsiveness. We now demonstrate that T cell recovery is gradual and is characterized by memory-type cell predominance and an increased proportion of CD4+ CD25+ CD127- FOXP3+ Treg during the lymphopenic period. Complete donor specific unresponsiveness in proliferative and cytotoxic assays, and in limiting dilution analyses of IL-2-producing and cytotoxic cells, developed and persisted for the 3-year follow-up in all patients, and extended to donor renal tubular epithelial cells. Assays in two of four patients were consistent with a role for a suppressive tolerance mechanism at 6 months to 1 year, but later (>= 18 months) studies on all four patients provided no evidence for a suppressive mechanism. Our studies demonstrate, for the first time, long-term, systemic donor-specific unresponsiveness in patients with HLA-mismatched allograft tolerance. While regulatory cells may play an early role, long-term tolerance appears to be maintained by a deletion or anergy mechanism. PMID- 21645256 TI - IL-17 expression by tubular epithelial cells in renal transplant recipients with acute antibody-mediated rejection. AB - Acute rejection is still a common complication of kidney transplantation. IL-17 is known to be associated with allograft rejection but the cellular source and the role of this cytokine remains unclear. We investigated IL-17 graft expression in renal transplant recipients with acute antibody-mediated rejection (ABMR), acute T-cell-mediated rejection (TCMR), interstitial fibrosis and tubular atrophy (IFTA) and acute tubular damage due to calcineurin-inhibitor toxicity (CNI). In acute ABMR, tubular IL-17 protein expression was significantly increased compared to TCMR, where most of the IL-17+ cells were CD4+ graft infiltrating lymphocytes, IFTA and CNI control groups. The tubular expression of IL-17 in acute ABMR colocalized with JAK2 phosphorylation and peritubular capillaries C4d deposition. In addition, IL-17 tubular expression was directly and significantly correlated with the extension of C4d deposits. In cultured proximal tubular cells, C3a induced IL-17 gene and protein expression along with an increased in JAK2 phosphorylation. The inhibition of JAK2 abolished C3a-induced IL-17 expression. The use of steroids and monoclonal antibodies reduced IL-17 expression, JAK2 phosphorylation and C4d deposition in acute ABMR patients. Our data suggest that tubular cells represent a significant source of IL-17 in ABMR and this event might be mediated by the complement system activation featuring this condition. PMID- 21645257 TI - Disclosing recipient information to potential living donors: preferences of donors and recipients, before and after surgery. AB - Consensus guidelines, while recommending that potential living donors should be given information that could impact their donation decision, are nonspecific about the types of information that should be disclosed. We surveyed potential (n = 36) and past (n = 45) living donors and transplant candidates (n = 45) and recipients (n = 45) about their preferences for sharing or knowing specific information about the recipient, how this information would impact decision making, and who should be responsible for disclosing information. Potential donors were less likely than all others to feel that recipient information should be disclosed to potential donors. Donors and recipients felt most strongly about disclosing if the recipient lost a previously transplanted kidney due to medication nonadherence as well as the likelihood of 1- and 5-year graft survival. Most donors would be less likely to pursue donation if the recipient lost a previously transplanted kidney due to medication nonadherence or generally had problems with taking medications as prescribed. Transplant programs should consider how to best balance the potential donor's right to receive information that could reasonably be expected to affect their decision-making process with the recipient's right to privacy and confidentiality. PMID- 21645258 TI - Early trajectories of depressive symptoms after liver transplantation for alcoholic liver disease predicts long-term survival. AB - Although it is well known that depression is associated with poorer medical outcomes, the association between depression- and liver transplant (LTX)-specific outcomes has not been investigated. We identified three trajectories of depressive symptoms evolving within the first post-LTX year in a cohort of 167 patients transplanted for alcoholic cirrhosis: a group with consistently low depression levels at all time points (group 1, n = 95), a group with initially low depression levels that rose over time (group 2, n = 41), and a group with consistently high depression levels (group 3, n = 31). Controlling for medical factors associated with poorer survival, recipients with increasing depression or persisting depression were more than twice as likely to die (all cause mortality) within the subsequent years. At 10 years post-LTX the survival rate was 66% for the low depression group, but only 46% and 43%, respectively, for the increasing depression and high depression groups. Except for a paradoxically higher percentage of malignancies in the low depression group, the causes of death and other specific LTX outcomes were not different between groups. Whether treatment of depression will improve survival rates is an area for research. PMID- 21645259 TI - The impact of IL28B genetic variants on recurrent hepatitis C in liver transplantation: significant lessons from a dual graft case. AB - IL28B genetic polymorphism is related to interferon-sensitivity in chronic hepatitis C, but the significance of grafts carrying different genotypes from recipients is still unclear in liver transplantation. A 51-year-old Japanese male carrying a minor genotype underwent dual liver transplantation for liver cirrhosis due to hepatitis C virus (HCV). The left lobe graft carried a major genotype, and the right a minor genotype. He achieved virological response during the course of pegylated-interferon and ribavirin therapy against recurrent hepatitis C for 2 years, but HCV relapsed immediately at the end of the therapy. Two years after antiviral therapy, liver biopsy was performed from each graft. The specimens showed A1F0 in the left lobe graft and A2F2 in the right. Moreover, quantitative polymerase chain reaction was performed using RNA extracted from each specimen to see there was no HCV RNA in the left lobe whereas there was in the right. This case provides clear evidence that IL28B genetic variants determine interferon sensitivity in recurrent hepatitis C following liver transplantation, which could result in new strategies for donor selection or for posttransplant antiviral therapy to HCV positive recipients. PMID- 21645260 TI - HIV transmitted from a living organ donor-New York City, 2009. PMID- 21645261 TI - Transmission of donor-specific skin condition from donor to recipient of facial allograft. PMID- 21645262 TI - Novel therapeutics based on recombinant botulinum neurotoxins to normalize the release of transmitters and pain mediators. AB - A major unmet clinical need exists for long-acting neurotherapeutics to alleviate chronic pain in patients unresponsive to available nonaddictive analgesics. Herein, a new strategy is described for the development of potent and specific inhibitors of the neuronal exocytosis of transmitters and pain mediators that exhibit unique antinociceptive activity. This entailed recombinant production in Escherichia coli of two serotypes of botulinum neurotoxin (BoNT) (BoNT(A) and BoNT(E) ), which are proteins that are known to block the release of transmitters by targeting and entering nerve endings, where their proteases cleave and inactivate a protein, synaptosomal protein of M(r) 25 000 (SNAP-25), that is essential for Ca(2+) -regulated exocytosis. Site-directed mutagenesis of Leu428 and Leu429 in BoNT(A) revealed that the remarkable longevity of its neuroparalytic action is attributable to a dileucine-containing motif. BoNT(E) acts transiently, because it lacks these residues, but is a superior inhibitor of transient receptor potential vanilloid type 1-mediated release of pain peptides from sensory nerves. The advantageous features of each serotype were harnessed by attaching the BoNT(E) protease moiety to an enzymically inactive mutant of BoNT(A) . The resultant purified composite protein could target motoneurons by binding to the BoNT(A) ectoacceptor and persistently produce BoNT(E) -truncated SNAP-25. As this enzyme lasted for more than 1 month (as compared with 5 days for BoNT(E) alone), such a dramatic extension in the lifetime of this BoNT(E) protease is attributable to a stabilizing influence of the BoNT(A) mutant. Most importantly, injecting this novel biotherapeutic into the foot pads of rats resulted in extended amelioration of inflammatory pain. Thus, a new generation of biotherapeutics has been created with the potential to give long-term relief of pain. PMID- 21645263 TI - Influence of antiseptics on microcirculation after neuronal and receptor blockade. AB - The topical application of the antiseptics octenidine and polyhexanide on wounds seems to improve microcirculation. These two antiseptics were tested in combination with neuronal inhibition and sympathethic receptor blockade to verify these findings, explore the influence of beta blockers on these microcirculative effects, and find out the principle of operation. Investigations were carried out on a standardised cremaster muscle model in rats (n = 66). The tested antiseptics, octenidine and polyhexanide were investigated alone (n = 12) and in combination with bupivacaine (n = 12), metoprolol (n = 12), phentolamine (n = 12) and surgical denervation (n = 12). Physiological saline was used for control (n = 6). The arteriolar diameter and functional capillary density (FCD) were investigated via trans-illumination microscopy before, as well as 60 and 120 minutes after application. Polyhexanide caused a significant increase in arteriolar diameter (86.5 +/- 3.8 um versus 100.0 +/- 3.6 um) and, like octenidine (7.2 +/- 0.7 n/0.22 mm(2) versus 11.6 +/- 0.6 n/0.22 mm(2) ), in FCD (9.2 +/- 0.5 versus 12.6 +/- 0.9) as well. When the antiseptics are used in combination with bupivacaine, metoprolol, phentolamine or surgical sympathectomy, these effects were eliminated or inverted. Assessing the results of the different blockades in combination with polyhexanide, we surmise that the antiseptic polyhexanide acts on the microcirculation mainly by blocking alpha receptors. This study shows that polyhexanide and octenidine improve muscular perfusion. Interestingly, the benefit of polyhexanide and octenidine on muscular perfusion is eliminated when the antiseptics are combined with other vasoactive agents, especially beta blockers. PMID- 21645264 TI - Diazepam blocks striatal lipid peroxidation and improves stereotyped activity in a rat model of acute stress. AB - In this work, the effect of a single dose of diazepam was tested on different markers of oxidative damage in the striatum of rats in an acute model of immobilization (restraint) stress. In addition, the locomotor activity was measured at the end of the restraint period. Immobilization was induced to animals for 24 hr, and then, lipid peroxidation, superoxide dismutase activity and content, and mitochondrial function were all estimated in striatal tissue samples. Corticosterone levels were measured in serum. Diazepam was given to rats as a pre-treatment (1 mg/kg, i.p.) 20 min. before the initiation of stress. Our results indicate that acute stress produced enhanced striatal levels of lipid peroxidation (73% above the control), decreased superoxide dismutase activity (54% below the control), reduced levels of mitochondrial function (35% below the control) and increased corticosterone serum levels (86% above the control). Pre treatment of stressed rats with diazepam decreased the striatal lipid peroxidation levels (68% below the stress group) and improved mitochondrial function (18% above the stress group), but only mild preservation of superoxide dismutase activity was detected (17% above the stress group). In regard to the motor assessment, only the stereotyped activity was increased in the stress group with respect to control (46% above the control), and this effect was prevented by diazepam administration (30% below the stress group). The preventive actions of diazepam in this acute model of stress suggest that drugs exhibiting anxiolytic and antioxidant properties might be useful for the design of therapies against early acute phases of physic stress. PMID- 21645265 TI - Hydroquinone stimulates inflammatory functions in microvascular endothelial cells via NF-kappaB nuclear activation. AB - Hydroquinone impairs several leucocyte cell functions, which alter the immune response. Although endothelial cell functions are important for the development of immune responses, hydroquinone actions on endothelial cell have not been shown. Therefore, the effect of hydroquinone exposure (10 or 100 MUM for 2 hr) on primary culture of microvascular endothelial cells (PMECs) obtained from the cremaster muscle of Wistar rats incubated in the presence or absence of lipopolysaccharide (LPS, 2 MUg/mL) was investigated. Hydroquinone treatment induced the membrane expression of cell adhesion molecules (CAMs) from the immunoglobulin superfamilies ICAM-1 (intercellular), VCAM-1(vascular) and PECAM-1 (platelet endothelial) and induced the secretion of cytokines interleukin-1beta (IL-1beta) and tumour necrosis factor-alpha (TNF-alpha). The effects were dependent on transcriptional modifications because enhanced CAM mRNA expression as well as both cytokines and nuclear factor kappaB (NF-kappaB) nuclear activation was found. These effects may be due to the direct action of hydroquinone rather than its quinone metabolites, because endothelial cells do not present myeloperoxidase enzyme and hydroquinone incubation did not induce the expression of cytochrome P450 2E1 (CYP2E1) or prostaglandin H synthase 1. In addition, the incubation of endothelial cells with benzoquinone (10 MUM, 2 hr) impaired PECAM-1 expression and did not modify NF-kappaB nuclear activation. Taken together, the data herein presented reveal that hydroquinone evokes pro inflammatory properties in endothelial cells that are triggered by the enhancement of NF-kappaB nuclear translocation-dependent gene transcription. PMID- 21645266 TI - P2X7 receptor gene polymorphism analysis in rheumatoid arthritis. AB - The P2X7 receptor, a member of the P2X family of nucleotide-gated channels, is predominantly expressed by monocytic cells. The activation of this receptor has been associated with downstream-signalling cascades, resulting in the release of a number of inflammatory mediators. There are more than 815 single nucleotide polymorphisms (SNPs) that have been described in the human P2X7R gene, but only few have been functionally characterized. The main aim of this study is to determine whether P2X7R gene polymorphisms confer susceptibility to rheumatoid arthritis (RA). A total of 125 patients with RA and 158 healthy volunteers were enrolled in this study. DNA fragment was PCR amplified and sequenced on the AB 3130 Genetic Analyzer. No significant difference in allele frequencies of 489 C >T, 1096 C->G and 1513 A->C polymorphisms, among sporadic cases of RA and healthy controls was found. However, the 1513A/C genotype was significantly associated with the presence of rheumatoid factor and anti-MCV autoantibody in RA patients. Interestingly, the genotype frequency of 1068 A/A was 0.19 in the RA group and 0.09 in control group (P = 0.025). Consequently, this polymorphism (AA) is two folds greater in the RA group compared to controls. Moreover, this polymorphism was significantly associated with mean concentration of C-reactive protein in RA patients. In contrast, 946G->A and 1729 T->A were not detected in both groups. As a result, these two polymorphisms are uncommon in Omani Arab population. Polymorphism at position 1068 and 1513 in the P2X7R gene might contribute to the pathogenesis of RA. Moreover, the loss-of-function SNP at position 1096 C->G or the gain-of-function SNP at position 489 C->T of the P2X7 gene does not appear to be a susceptibility gene locus for the development of RA. Further studies are required to confirm this finding. PMID- 21645267 TI - N-terminal probrain natriuretic peptide as a biomarker of cardioembolic stroke. AB - BACKGROUND: and purpose N-terminal probrain natriuretic peptide, which is mainly produced by the heart, is increased in acute stroke. We aimed to determine if N terminal probrain natriuretic peptide could be a biomarker for ischemic stroke with a cardioembolic cause. METHODS: Consecutive sample of acute stroke patients admitted to a Stroke Unit. Ischemic stroke subtype was classified using the TOAST classification. Blood samples were drawn within 72 h after stroke onset. Serum N terminal probrain natriuretic peptide concentration was measured using an electrochemiluminescence immunoassay. Mean values of N-terminal probrain natriuretic peptide were compared between patients with hemorrhagic stroke vs. ischemic stroke, cardioembolic stroke vs. noncardioembolic stroke, cardioembolic stroke with atrial fibrillation vs. noncardioembolic stroke using t-test. Receiver operating characteristic curves were used to test the ability of N terminal probrain natriuretic peptide values to identify cardioembolic stroke and cardioembolic stroke with atrial fibrillation. RESULTS: Ninety-two patients were included (66 with ischemic stroke) with a mean age of 58.6 years. Twenty-eight (42.4%) ischemic strokes had a cardioembolic cause. Mean N-terminal probrain natriuretic peptide values for cardioembolic stroke were significantly higher (P<0.001) (491.6; 95% confidence interval 283.7-852.0 pg/ml) than for noncardioembolic ischemic stroke (124.7; 86.3-180.2 pg/ml). The area under the receiver operating characteristic curve for N-terminal probrain natriuretic peptide in cardioembolic stroke was 0.77. The cut-off point with the highest sensitivity and specificity was set at 265.5 pg/ml (71.4% and 73.7% respectively). The area under the curve of N-terminal probrain natriuretic peptide for cardioembolic stroke related to atrial fibrillation was 0.92, cut-off was set at 265.5 pg/ml (sensitivity 94.4%, specificity 72.9%). CONCLUSION: N terminal probrain natriuretic peptide is a biomarker with a good accuracy to predict ischemic stroke of cardioembolic cause, namely associated with atrial fibrillation. PMID- 21645268 TI - Outcomes in acute ischemic strokes presenting with disabling neurologic deficits without intracranial vascular occlusion. AB - BACKGROUND: Patients with moderate to severe acute ischemic stroke without intracranial vessel occlusion are an intriguing subset of stroke patients. They pose diagnostic and therapeutic challenges to the physician. We sought to study these patients with an emphasis on their radiological and clinical outcomes. METHODS: This is a retrospective cohort study of ischemic stroke patients (NIHSS>=6), with no intracranial vessel occlusion on computed tomography angiography within six-hours of symptom onset. Follow-up imaging - either computed tomography brain or magnetic resonance imaging - was performed within one- to seven-days. The primary outcome was modified Rankin Scale score<=2 at three-months. RESULTS: In a database of 1308 patients, we identified 99 (7.6%) patients with NIHSS>=6 and no intracranial vessel occlusion on computed tomography angiography. The mean age was 67.8 +/- 15.4 years and 60 (60.6%) were men. The median baseline NIHSS was nine (6-28). The initial computed tomography head was normal in 79 (79.8%) patients. Dramatic early clinical improvement at 24 h (NIHSS score <=2 at 24 h or change between baseline and 24 h NIHSS score >=15 points) was seen in 38 (38.4%) patients. Follow-up scans showed infarcts in 66 (66.7%) patients. Fifty (50.5%) patients received tissue plasminogen activator; one (2%) tissue plasminogen activator-treated patient developed symptomatic intracranial hemorrhage. At three-months; 59 (59.6%) patients were independent (modified Rankin Scale<=2), 34 (34.3%) patients were dependent (modified Rankin Scale 3-5), and six (6.1%) were dead. The factors associated with the unlikelihood of good outcome were higher initial NIHSS (odds ratio 0.86 per additional point, 95% confidence interval 0.77-0.95, P=0.003), and older age (odds ratio 0.95 per additional year, 95% confidence interval 0.92-0.98, P=0.004). CONCLUSION: Stroke without intracranial occlusions are not a benign entity. Factors that are independently associated with decreased likelihood of a good outcome are higher baseline NIHSS, and older age. Treatment with tissue plasminogen activator is not a predictor of outcome. PMID- 21645269 TI - The Membrane-Activated Chelator Stroke Intervention (MACSI) Trial of DP-b99 in acute ischemic stroke: a randomized, double-blind, placebo-controlled, multinational pivotal phase III study. AB - RATIONALE: Zinc is both a direct neurotoxin and a signaling mediator in multiple early and late detrimental processes following ischemia. DP-b99, a lipophilic moderate-affinity chelator of zinc, is a first-in-class multitargeted neuroprotective agent for ischemic stroke. DP-b99 has completed several Phase I studies and two double-blind placebo-controlled Phase II trials, which supported the safety of DP-b99 and were consistent with a beneficial effect on poststroke recuperation. AIM: Membrane-Activated Chelator Stroke Intervention is a Phase III study. The primary objective is to evaluate the safety and therapeutic effects of intravenous 1.0 mg/kg/day DP-b99, initiated within nine-hours of stroke onset in patients with moderately severe hemispheric acute ischemic stroke, through the analysis across the whole distribution of scores of the primary efficacy endpoint of the modified Rankin Scale, 90 days after the stroke. METHODS: The Membrane Activated Chelator Stroke Intervention study is a randomized, double-blind, placebo-controlled, multicenter, multinational, parallel-arm trial comparing a placebo group to a group treated with intravenous DP-b99 for four consecutive days. Non-rtPA-treated acute ischemic stroke patients--with a baseline NIHSS score of 10-16 and a clinical syndrome that includes language dysfunction, visual field defect and/or neglect--will be stratified on a 1:1 basis to one of the two treatments. Half will be randomized within 0-4.5 h of stroke onset. Follow-up after the four treatment days will occur on days 12, 30 and 90. An interim futility analysis will be performed after primary endpoint data have been collected for 50% of 770 subjects planned to be enrolled. A data and safety monitoring board will assess safety data and will oversee the interim analysis. CONCLUSION: This Phase III Membrane-Activated Chelator Stroke Intervention trial is based on promising data derived from previous Phase I and II DP-b99 trials and capitalizes on lessons learned from failures of past stroke studies in relation to neuroprotection, patient selection and data analysis. PMID- 21645270 TI - Red cell distribution width does not predict stroke severity or functional outcome. AB - INTRODUCTION: Red cell distribution width was recently identified as a predictor of cardiovascular and all-cause mortality in patients with previous stroke. Red cell distribution width is also higher in patients with stroke compared with those without. However, there are no data on the association of red cell distribution width, assessed during the acute phase of ischemic stroke, with stroke severity and functional outcome. In the present study, we sought to investigate this relationship and ascertain the main determinants of red cell distribution width in this population. METHODS: We used data from the Acute Stroke Registry and Analysis of Lausanne for patients between January 2003 and December 2008. Red cell distribution width was generated at admission by the Sysmex XE-2100 automated cell counter from ethylene diamine tetraacetic acid blood samples stored at room temperature until measurement. An chi(2) -test was performed to compare frequencies of categorical variables between different red cell distribution width quartiles, and one-way analysis of variance for continuous variables. The effect of red cell distribution width on severity and functional outcome was investigated in univariate and multivariate robust regression analysis. Level of significance was set at 95%. RESULTS: There were 1504 patients (72+/-15.76 years, 43.9% females) included in the analysis. Red cell distribution width was significantly associated to NIHSS (beta-value=0.24, P=0.01) and functional outcome (odds ratio=10.73 for poor outcome, P<0.001) at univariate analysis but not multivariate. Prehospital Rankin score (beta=0.19, P<0.001), serum creatinine (beta=0.008, P<0.001), hemoglobin (beta=-0.009, P<0.001), mean platelet volume (beta=0.09, P<0.05), age (beta=0.02, P<0.001), low ejection fraction (beta=0.66, P<0.001) and antihypertensive treatment (beta=0.32, P<0.001) were independent determinants of red cell distribution width. CONCLUSIONS: Red cell distribution width, assessed during the early phase of acute ischemic stroke, does not predict severity or functional outcome. PMID- 21645271 TI - Calculation of numbers-needed-to-treat in parallel group trials assessing ordinal outcomes: case examples from acute stroke and stroke prevention. AB - BACKGROUND: Number-needed-to-treat describes the magnitude of the effect of an intervention, underpins health economic analyses, and is typically calculated for binary events. Ordered categorical outcomes provide more clinical information and their analysis using ordinal approaches is usually more efficient statistically. However, to date, techniques to calculate number-needed-to-treat based on ordinal outcomes for parallel group trials have had important limitations. Aims Numbers needed-to-treat may be calculated for ordinal data from parallel group trials by using an unmatched comparison of all subjects or by generating matched pairs of patients nested within the study. METHODS: The above approaches were assessed and compared with numbers-needed-to-treat calculated for binary outcomes using individual patient data from acute and prevention stroke trials testing the effect of interventions of varying utility and efficacy. RESULTS: Numbers-needed to-treat were generally lower numerically for ordinal vs. binary, and matched vs. unmatched analyses, and the lowest in highly efficacious interventions: hemicraniectomy, ordinal matched 2.4 vs. ordinal unmatched 2.5 vs. binary matched 12 vs. binary unmatched 9 (one trial, 12 month outcome); alteplase, 4.5 vs. 6.6 vs. 8.4 vs. 8.4 (one trial with two parts, three-months); aspirin, 42 vs. 58 vs. 76 vs. 80 (one trial, six-months); and stroke units, 3.6-5.3 vs. 6.2 vs. 4.7-5.9 vs. 6.3-7.0 (two trials, three- to 60 months). Similar trends were seen for aspirin/dipyridamole vs. aspirin in secondary prevention, 22 vs. 20 vs. 31 vs. 31 (one trial, 24 months). CONCLUSIONS: Number-needed-to-treat may be calculated for ordinal outcome data derived from parallel group stroke trials; such numbers needed-to-treat are lower than those calculated for binary outcomes. Their use complements the use of ordinal statistical approaches in the analysis of ordered categorical data. PMID- 21645272 TI - An integrative look at frugivory and seed dispersal studies. PMID- 21645273 TI - Effect of logging on rodent scatter-hoarding dynamics in tropical forests: implications for plant recruitment. AB - The present study tested the hypothesis that logging affects the scatter-hoarding behavior of rodents, which, in turn, negatively affects the quantity and quality of Pouteria campechiana (Sapotaceae) seed dispersal. A series of seed stations was established in logged and unlogged forests of ejido Senor, Yucatan Peninsula, and comparisons were made between logged and unlogged forests in terms of: (i) seed removal; (ii) number of seeds hoarded; (iii) hoarding distance; and (iv) the number of recruits and the survival of hoarded seeds. The number of both hoarded and removed seeds was significantly higher in unlogged sites. Furthermore, the mean distance of hoarding was greater in unlogged compared with logged sites. Although recruitment and survival were present in both logged and unlogged sites, there were more surviving seedlings in unlogged sites. The data indicate that both the quantity and quality of seed dispersal are negatively affected by logging because of a change in the rodent scatter-hoarding dynamics. These changes suggest that plant-animal interactions are crucial to the understanding of the ecology and conservation of managed tropical forests. PMID- 21645274 TI - Habitat differences in dung beetle assemblages in an African savanna-forest ecotone: implications for secondary seed dispersal. AB - The probability and pattern of secondary seed dispersal by dung beetles (Scarabaeinae) depend on their community structure and composition at the site of primary deposition, which, in turn, seem to be strongly determined by vegetation. Consequently, we expected pronounced differences in secondary seed dispersal between forest and savanna in the northern Ivory Coast, West Africa. We found 99 dung beetle species at experimentally exposed dung piles of the olive baboon (Papio anubis (Lesson, 1827)), an important primary seed disperser in West Africa. Seventy-six species belonged to the roller and tunneler guilds, which are relevant for secondary seed dispersal. Most species showed a clear habitat preference. Contrary to the Neotropics, species number and abundance were much higher in the savanna than in the forest. Rollers and tunnelers each accounted for approximately 50% of the individuals in the savanna, but in the forest rollers made up only 4%. Seeds deposited into the savanna by an omnivorous primary disperser generally have a higher overall probability of being more rapidly dispersed secondarily by dung beetles than seeds in the forest. Also, rollers disperse seeds over larger distances. In contrast to other studies, small rollers were active in dispersal of large seeds, which were seemingly mistaken for dung balls. Our results suggest that rollers can remove seeds from any plant dispersed in primate dung in this ecosystem. PMID- 21645275 TI - Seed dispersal in Hong Kong, China: past, present and possible futures. AB - In the present article, published and unpublished information regarding seed dispersal in the degraded landscape of Hong Kong, China, is reviewed. Information was available for 1681 native plant species, of which 1165 were assigned to probable seed dispersal modes. Endozoochory accounted for one-third (34.4%) of all species evaluated, half (54.4%) of those from forests, and more than two thirds (69.2%) of all trees and tall shrubs. Wind dispersal (25.9%) and dispersal by an unknown agent (30.7%) accounted for most of the rest, with the unknown species mostly small-seeded herbs. Although the frugivore fauna of the Hong Kong region has been truncated since the late Pleistocene, there are few clear examples of failed mutualisms. The most striking is the absence of scatter hoarding rodents from Hong Kong, despite the presence of forest trees that appear to require them for effective dispersal. There are also some large Lauraceae fruits that appear to be targeted at larger-gaped birds than currently present. Most endozoochorous species are dispersed by 3 small passerine birds (Pycnonotus jocosus, P. sinensis, and Zosterops japonicus), with larger birds, fruit bats (Cynopterus sphinx, Rousettus leschenaulti), rhesus macaques (Macaca mulatta), civets (Paguma larvata, Viverricula indica), and muntjacs (Muntiacus muntjac) accounting for the remainder. The low plant diversity in frugivore droppings, seed traps, and secondary vegetation suggests plant succession may be dispersal limited, although this has not been investigated experimentally. Planting underdispersed species is the simplest solution but, in the longer term, the (re)introduction of a scatter-hoarding rodent should be considered as well as that of other locally and regionally extinct frugivores. PMID- 21645276 TI - Frugivory and seed dispersal in the Galapagos: what is the state of the art? AB - The Galapagos are considered a model oceanic archipelago, with unique flora and fauna currently threatened by alien invasive species. Seed dispersal is an important ecosystem function with consequences for plant population dynamics and vegetation structure. Hence, understanding the seed dispersal abilities of the assemblages of frugivores will inform scientists and managers of the dynamics of plant invasions and improve management planning. Here we provide the first comprehensive review of published information on frugivory and animal seed dispersal in the Galapagos. We collected data from a variety of sources, including notes of the first naturalist expeditions, gray literature available only in Galapagos collections, and peer-reviewed journal articles. Plant-animal frugivorous interactions were retrieved from 43 studies and compiled into an interaction matrix describing 366 unique interactions. Most studies focused on fruit consumption as a driving force for natural selection, but seed fate was seldom considered. Although most (71%) of the interactions involved native plants, more than one-quarter (28%) involved introduced species. Interactions involving birds are considerably more common than those of reptiles and mammals, probably reflecting a research bias towards birds. Despite the historical importance of the archipelago as the laboratory for evolutionary and ecological research, understanding of its seed dispersal systems is limited. We end the review by suggesting 3 priority areas of research on frugivory and seed dispersal in the Galapagos: (i) target research to close knowledge gaps; (ii) the use of a network approach to frame seed dispersal at the community level; and (iii) evaluation of the effect of seed dispersal as a selective pressure acting upon plants and frugivores. Finally, the output of this research has to be properly delivered to the Galapagos National Park Services to help increase management effectiveness. PMID- 21645277 TI - Relationships among rat numbers, abundance of oil palm fruit and damage levels to fruit in an oil palm plantation. AB - The relationships between vertebrate pests and crop damage are often complex and difficult to study. In palm oil plantations rodents remain the major pests, causing substantial monetary losses. The present study examined the numerical and functional responses of rodents to changes in the availability of oil palm fruit and the damage associated with that response. For the study, 200 traps were set in pairs on a 10 * 10 trapping grid for 3 consecutive nights in each of 6 study plots at 8-week intervals in a 2569 ha oil palm plantation at Labu, Negeri Sembilan state in Peninsular Malaysia over 14 months. A total of 1292 individual rats were captured over 25 200 trap-nights. Animals were identified, aged, sexed, weighed and measured. An index of the relative abundance of rats was calculated based on trapping success. Damage to infructescences was assessed at each trap point. Regardless of the age of palms, there were positive and significant relationships between the relative abundance of rats and numbers of infructescences. The levels of damage to infructescences were significantly correlated with the relative abundance of rats. A steep increase in damage was observed with an increase in mature infructescences, indicating a feeding preference of rats for mature infructescences. For both males and females of all rat species, there were weak and non-significant correlations between body condition and infructescence numbers. These results indicated that there was a numerical and a functional response by rats to the availability of palm fruit and a resulting increase in depredation of oil palm fruits. The ways in which this information might aid in future pest control are discussed. PMID- 21645278 TI - Oxidative stress physiology in relation to life history traits of a free-living vertebrate: the spotted snow skink, Niveoscincus ocellatus. AB - Recent research suggests that oxidative stress, via its links to metabolism and senescence, is a key mechanism linking life history traits such as fecundity and growth with survival; however, this has rarely been put under empirical scrutiny within free-living populations. Using a wild population of live-bearing skinks, we explored how plasma antioxidant activity (OXY), reactive oxidative metabolites (ROM), and the estimated oxidative stress index are associated with female and male life history. We found that male skinks have a significantly higher ROM and estimated oxidative stress index than female skinks, but this was not accompanied by a sex difference in mortality. Both sexes showed a non-linear association between OXY and age, indicating that the oldest and youngest individuals had the lowest OXY. Interestingly, female skinks with high OXY showed a decreased probability of survival to the following season. However, we found no significant associations between female reproductive investment (litter size or litter mass) or parturition date (i.e. metabolism) and oxidative status. Combined, our results offer mixed support for a role of oxidative stress in mediating life history traits and suggest that future studies need to explore oxidative stress during vitellogenesis in addition to using an intra-individual approach to understand the cost of reproduction and patterns of aging. PMID- 21645279 TI - Blood electrolytes of the freshwater catfish Heteropneustes fossilis in response to treatment with a botanical pesticide (latex of Euphorbia royleana). AB - The aim of the present study was to determine changes in blood electrolytes (calcium and phosphate) in the freshwater catfish Heteropneustes fossilis following short- and long-term exposure to latex of Euphorbia royleana. To investigate the effects of short-term exposure, fish were exposed to 80% of the 96-h LC(50) of E. royleana latex (2.47 mg/L) for 96 h. To assess the effects of long-term exposure, fish were exposed to 20% of the 96-h LC(50) of E. royleana latex (0.618 mg/L) for 28 days. Fish were killed after 24, 48, 72 and 96 h (short term experiments) or after 7, 14, 21 and 28 days (long-term experiments). Blood samples were collected and sera were analyzed to determine calcium and inorganic phosphate levels. Serum calcium levels decreased progressively in H. fossilis after 48 h exposure to E. royleana latex. This decrease persisted until the end of the experiment (96 h). Serum inorganic phosphate levels in treated fish decreased progressively from 72 h. After 7 days exposure to E. royleana latex, decreased serum calcium levels were noted in H. fossilis that persisted until the end of the experiment. Serum phosphate levels were decreased in H. fossilis on days 21 and 28 of exposure to E. royleana latex. It is concluded that E. royleana latex alters the blood electrolyte content of the fish. The change in these electrolytes, particularly calcium levels, may cause disturbances in the normal vital physiological functions of the fish, its growth rate, and even its ability to survive in nature. PMID- 21645280 TI - Ranging behavior of the Francois' langur (Trachypithecus francoisi) in limestone habitats of Nonggang, China. AB - Ranging behavior is an important aspect of animal behavior that researchers use to investigate ecological influences on individual behavior. We collected data on the ranging behavior of one group of Francois' langurs (Trachypithecus francoisi) between August 2003 and July 2004 in a limestone habitat within Nonggang Nature Reserve, China. We evaluated the influences of food availability, water resources and sleeping sites on ranging behavior. During the study period, the total home range size for the study group was 64.5 or (69.3 ha if the lacunae within the borders in which langurs were not observed were included), and the mean daily path length was 541 m. These are well within the range of variation reported in Trachypithecus species. The monthly range size was between 9.8 and 23.3 ha, and monthly range size correlated negatively with young leaf availability. The langurs tended to use a larger range size during young leaf-lean periods. The langurs did not use their home range uniformly, and 74% of their activities occurred within 35% of their home range. The most heavily used quadrats in the home range were located near the most frequently used sleeping sites, suggesting that sleeping sites have a significant influence on the ranging behavior of Francois' langurs in limestone habitats. PMID- 21645281 TI - Complete blood count using VCS (volume, conductivity, light scatter) technology is affected by hyperlipidemia in a child with acute leukemia. AB - Asparaginase, an effective drug in the treatment of childhood acute lymphoblastic leukemia (ALL), has become an important component of most childhood ALL regimens during the remission induction or intensification phases of treatment. The incidence range of asparaginase-associated lipid abnormalities that are seen in children is 67-72%. Lipemia causes erroneous results, which uses photometric methods to analyze blood samples. We describe a case of l-asparaginase-associated severe hyperlipidemia with complete blood count abnormalities. Complete blood count analysis was performed with Beckman COULTER((r)) GEN.STM system, which uses the Coulter Volume, Conductivity, Scatter technology to probe hydrodynamically focused cells. Although an expected significant inaccuracy in hemoglobin determination occurred starting from a lipid value of 3450 mg/dl, we observed that triglyceride level was 1466 mg/dl. Complete blood count analysis revealed that exceptionally high hemoglobin, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration levels vs. discordant with red blood cell count, mean corpuscular volume, and hematocrit levels. Total leukocyte count altered spontaneously in a wide range, and was checked with blood smear. Platelet count was in expected range (Table 1). Thus, we thought it was a laboratory error, and the patient's follow-up especially for red cell parameters was made by red blood cell and hematocrit values. PMID- 21645282 TI - Serum levels of granulocyte colony-stimulating factor in JAK2 V617F-positive vs. negative erythrocytosis. PMID- 21645283 TI - D-dimer level in pediatric patients with solid and hematologic malignancies, Shiraz, Southern Iran. PMID- 21645284 TI - Neuroprotective effects of tempol acyl esters against retinal ganglion cell death in a rat partial optic nerve crush model. AB - PURPOSE: The aim of this study is to search for more effective derivatives of the superoxide dismutase mimetic tempol (4-hydroxy-2,2,6,6-tetramethylpiperidine-1 oxyl). Although tempol is neuroprotective in a rat partial optic nerve crush (PONC) model, relatively high doses are required to exert this effect. METHODS: Tempol acyl esters with different-length fatty acids (tempol-C4, tempol-C8, tempol-C12 and tempol-C16) were synthesized and the following properties were evaluated: water-octanol partition coefficient, liposome-liposome energy transfer, and electron paramagnetic resonance (EPR). Brown Norway rats underwent PONC and received tempol or acyl esters intraperitoneally once daily for 7 consecutive days. We then compared the effects of tempol and its four esters on retinal ganglion cell (RGC) damage using a retrograde labelling method. RESULTS: The water-octanol partition coefficient increased with increasing length of attached acyl chain. However, the energy of the liposome-liposome transfer seemed to be optimal for tempol-C8 and tempol-C12. The EPR signal was very similar for all tested compounds, suggesting similar efficiency of superoxide scavenging. Partial optic nerve crush in vehicle-treated animals reduced RGC numbers by approx. 59% when compared with sham-operated eyes. Tempol did not affect RGC loss at a dose of 1 mg/kg. In contrast, at molar doses equivalent to 1 mg/kg of tempol, tempol-C8 showed a significant neuroprotective effect, whereas tempol-C4, tempol-C12 and tempol-C16 did not act neuroprotectively. CONCLUSION: Manipulating the hydrophobicity of tempol seems to be a promising tool for developing more potent neuroprotectants in the PONC degeneration model. However, the resulting compounds need further pharmacological evaluation. PMID- 21645286 TI - Risk for highly pathogenic avian influenza H5N1 virus infection in chickens in small-scale commercial farms, in a high-risk area, Bangladesh, 2008. AB - Small-scale commercial chicken farms (FAO-defined system 3) with poor biosecurity predominate in developing countries including Bangladesh. By enroling fifteen highly pathogenic avian influenza (HPAI) cases occurring in such farms - (February - April 2008) and 45 control farms (March-May 2008) with similar set up, we conducted a case-control study to evaluate the risk factors associated with HPAI H5N1 virus infections in chickens reared in small-scale commercial farms in a spatially high-risk area in Bangladesh. Data collected by a questionnaire from the selected farms were analysed by univariable analysis and multivariable conditional logistic regression. The risk factors independently associated were 'dead crow seen at or near farm' [odds ratio (OR) 47.4, 95% confidence interval (CI) 4.7-480.3, P = 0.001], 'exchanging eggtrays with market vendors' (OR 20.4, 95% CI 1.9-225.5, P = 0.014) and 'mortality seen in backyard chicken reared nearby' (OR 19.4, 95% CI 2.8-131.9, P = 0.002). These observations suggest that improved biosecurity might reduce the occurrence of HPAI outbreaks in small-scale commercial farms in Bangladesh. PMID- 21645287 TI - Prevalence of HBV and HBV vaccination coverage in health care workers of tertiary hospitals of Peshawar, Pakistan. AB - BACKGROUND: Hepatitis B Virus (HBV) may progress to serious consequences and increase dramatically beyond endemic dimensions that transmits to or from health care workers (HCWs) during routine investigation in their work places. Basic aim of this study was to canvass the safety of HCWs and determine the prevalence of HBV and its possible association with occupational and non-occupational risk factors. Hepatitis B vaccination coverage level and main barriers to vaccination were also taken in account. RESULTS: A total of 824 health care workers were randomly selected from three major hospitals of Peshawar, Khyber Pakhtunkhwa. Blood samples were analyzed in Department of Zoology, Kohat University of Science and Technology Kohat, and relevant information was obtained by means of preset questionnaire. HCWs in the studied hospitals showed 2.18% prevalence of positive HBV. Nurses and technicians were more prone to occupational exposure and to HBV infection. There was significant difference between vaccinated and non-vaccinated HCWs as well as between the doctors and all other categories. Barriers to complete vaccination, in spite of good knowledge of subjects in this regard were work pressure (39.8%), negligence (38.8%) un-affordability (20.9%), and unavailability (0.5%). CONCLUSIONS: Special preventive measures (universal precaution and vaccination), which are fundamental way to protect HCW against HBV infection should be adopted. PMID- 21645288 TI - Report from the second international symposium on animal genomics for animal health: critical needs, challenges and potential solutions. AB - The second International Symposium on Animal Genomics for Animal Health held in Paris, France 31 May-2 June, 2010, assembled more than 140 participants representing research organizations from 40 countries. The symposium included a roundtable discussion on critical needs, challenges and opportunities, and a forward look at the potential applications of animal genomics in animal health research. The aim of the roundtable discussion was to foster a dialogue between scientists working at the cutting edge of animal genomics research and animal health scientists. Importantly, stakeholders were included to provide input on priorities and the potential value of animal genomics to the animal health community. In an effort to facilitate the roundtable discussion, the organizers identified four priority areas to advance the use of genome-enabled technologies in animal health research. Contributions were obtained through open discussions and a questionnaire distributed at the start of the symposium. This report provides the outcome of the roundtable discussion for each of the four priority areas. For each priority, problems are identified, including potential solutions and recommendations. This report captures key points made by symposium participants during the roundtable discussion and serves as a roadmap to steer future research priorities in animal genomics research. PMID- 21645289 TI - Transcription variants of SLA-7, a swine non classical MHC class I gene. AB - In pig, very little information is available on the non classical class I (Ib) genes of the Major Histocompatibility Complex (MHC) i.e. SLA-6, -7 and -8. Our aim was to focus on the transcription pattern of the SLA-7 gene. RT-PCR experiments were carried out with SLA-7 specific primers targeting either the full coding sequence (CDS) from exon 1 to the 3 prime untranslated region (3UTR) or a partial CDS from exon 4 to the 3UTR. We show that the SLA-7 gene expresses a full length transcript not yet identified that refines annotation of the gene with eight exons instead of seven as initially described from the existing RefSeq RNA. These two RNAs encode molecules that differ in cytoplasmic tail length. In this study, another SLA-7 transcript variant was characterized, which encodes a protein with a shorter alpha 3 domain, as a consequence of a splicing site within exon 4. Surprisingly, a cryptic non canonical GA-AG splicing site is used to generate this transcript variant. An additional SLA-7 variant was also identified in the 3UTR with a splicing site occurring 31 nucleotides downstream to the stop codon. In conclusion, the pig SLA-7 MHC class Ib gene presents a complex transcription pattern with two transcripts encoding various molecules and transcripts that do not alter the CDS and may be subject to post-transcriptional regulation. PMID- 21645290 TI - Gene expression pattern in swine neutrophils after lipopolysaccharide exposure: a time course comparison. AB - BACKGROUND: Experimental exposure of swine neutrophils to bacterial lipopolysaccharide (LPS) represents a model to study the innate immune response during bacterial infection. Neutrophils can effectively limit the infection by secreting lipid mediators, antimicrobial molecules and a combination of reactive oxygen species (ROS) without new synthesis of proteins. However, it is known that neutrophils can modify the gene expression after LPS exposure. We performed microarray gene expression analysis in order to elucidate the less known transcriptional response of neutrophils during infection. METHODS: Blood samples were collected from four healthy Iberian pigs and neutrophils were isolated and incubated during 6, 9 and 18 hrs in presence or absence of lipopolysaccharide (LPS) from Salmonella enterica serovar Typhimurium. RNA was isolated and hybridized to Affymetrix Porcine GeneChip(r). Microarray data were normalized using Robust Microarray Analysis (RMA) and then, differential expression was obtained by an analysis of variance (ANOVA). RESULTS: ANOVA data analysis showed that the number of differentially expressed genes (DEG) after LPS treatment vary with time. The highest transcriptional response occurred at 9 hr post LPS stimulation with 1494 DEG whereas at 6 and 18 hr showed 125 and 108 DEG, respectively. Three different gene expression tendencies were observed: genes in cluster 1 showed a tendency toward up-regulation; cluster 2 genes showing a tendency for down-regulation at 9 hr; and cluster 3 genes were up-regulated at 9 hr post LPS stimulation. Ingenuity Pathway Analysis revealed a delay of neutrophil apoptosis at 9 hr. Many genes controlling biological functions were altered with time including those controlling metabolism and cell organization, ubiquitination, adhesion, movement or inflammatory response. CONCLUSIONS: LPS stimulation alters the transcriptional pattern in neutrophils and the present results show that the robust transcriptional potential of neutrophils under infection conditions, indicating that active regulation of gene expression plays a major role in the neutrophil-mediated- innate immune response. PMID- 21645291 TI - Analysis of global transcriptional responses of chicken following primary and secondary Eimeria acervulina infections. AB - BACKGROUND: Characterization of host transcriptional responses during coccidia infections can provide new clues for the development of alternative disease control strategies against these complex protozoan pathogens. METHODS: In the current study, we compared chicken duodenal transcriptome profiles following primary and secondary infections with Eimeria acervulina using a 9.6K avian intestinal intraepithelial lymphocyte cDNA microarray (AVIELA). RESULTS: Gene Ontology analysis showed that primary infection significantly modulated the levels of mRNAs for genes involved in the metabolism of lipids and carbohydrates as well as those for innate immune-related genes. By contrast, secondary infection increased the levels of transcripts encoded by genes related to humoral immunity and reduced the levels of transcripts for the innate immune-related genes. The observed modulation in transcript levels for gene related to energy metabolism and immunity occurred concurrent with the clinical signs of coccidiosis. CONCLUSIONS: Our results suggest that altered expression of a specific set of host genes induced by Eimeria infection may be responsible, in part, for the observed reduction in body weight gain and inflammatory gut damage that characterizes avian coccidiosis. PMID- 21645292 TI - Cross reactive cellular immune responses in chickens previously exposed to low pathogenic avian influenza. AB - BACKGROUND: Avian influenza (AI) infection in poultry can result in high morbidity and mortality, and negatively affect international trade. Because most AI vaccines used for poultry are inactivated, our knowledge of immunity against AI is based largely on humoral immune responses. In fact, little is known about cellular immunity following a primary AI infection in poultry, especially regarding cytotoxic T lymphocytes (CTL's). METHODS: In these studies, major histocompatibility complex (MHC)-defined (B2/B2) chickens were infected with low pathogenic AI (LPAI) H9N2 and clinical signs of disease were monitored over a two weeks period. Splenic lymphocytes from infected and naive birds were examined for cross reactivity against homologous and heterologous (H7N2) LPAI by ex vivo stimulation. Cellular immunity was determined by cytotoxic lysis of B2/B2 infected lung target cells and proliferation of T cells following exposure to LPAI. RESULTS: Infection with H9N2 resulted in statistically significant weight loss compared to sham-infected birds. Splenic lymphocytes derived from H9N2 infected birds displayed lysis of both homologous (H9N2) and heterologous (H7N2) infected target cells, whereas lymphocytes obtained from sham-infected birds did not. T cell proliferation was determined to be highest when exposed to the homologous virus. CONCLUSIONS: Taken together these data extend the findings that cellular immunity, including CTL's, is cross reactive against heterologous isolates of AI and contribute to protection following infection. PMID- 21645293 TI - Genome-wide identification of allele-specific expression (ASE) in response to Marek's disease virus infection using next generation sequencing. AB - BACKGROUND: Marek's disease (MD), a T cell lymphoma induced by the highly oncogenic alpha-herpesvirus Marek's disease virus (MDV), is the main chronic infectious disease concern threatening the poultry industry. Enhancing genetic resistance to MD in commercial poultry is an attractive method to augment MD vaccines, which is currently the control method of choice. In order to optimally implement this control strategy through marker-assisted selection (MAS) and to gain biological information, it is necessary to identify specific genes that influence MD incidence. METHODS: A genome-wide screen for allele-specific expression (ASE) in response to MDV infection was conducted. The highly inbred ADOL chicken lines 6 (MD resistant) and 7 (MD susceptible) were inter-mated in reciprocal crosses and half of the progeny challenged with MDV. Splenic RNA pools at a single time after infection for each treatment group point were generated, sequenced using a next generation sequencer, then analyzed for allele-specific expression (ASE). To validate and extend the results, Illumina GoldenGate assays for selected cSNPs were developed and used on all RNA samples from all 6 time points following MDV challenge. RESULTS: RNA sequencing resulted in 11-13+ million mappable reads per treatment group, 1.7+ Gb total sequence, and 22,655 high-confidence cSNPs. Analysis of these cSNPs revealed that 5360 cSNPs in 3773 genes exhibited statistically significant allelic imbalance. Of the 1536 GoldenGate assays, 1465 were successfully scored with all but 19 exhibiting evidence for allelic imbalance. CONCLUSIONS: ASE is an efficient method to identify potentially all or most of the genes influencing this complex trait. The identified cSNPs can be further evaluated in resource populations to determine their allelic direction and size of effect on genetic resistance to MD as well as being directly implemented in genomic selection programs. The described method, although demonstrated in inbred chicken lines, is applicable to all traits in any diploid species, and should prove to be a simple method to identify the majority of genes controlling any complex trait. PMID- 21645294 TI - Evidence for genetic variance in resistance to tuberculosis in Great Britain and Irish Holstein-Friesian populations. AB - BACKGROUND: Here, we jointly summarise scientific evidence for genetic variation in resistance to infection with Mycobacterium bovis, the primary agent of bovine tuberculosis (TB), provided by two recent and separate studies of Holstein Friesian dairy cow populations in Great Britain (GB) and Ireland. METHODS: The studies quantified genetic variation within archived data from field and abattoir surveillance control programmes within each country. These data included results from the single intradermal comparative tuberculin test (SICTT), abattoir inspection for TB lesions and laboratory confirmation of disease status. Threshold animal models were used to estimate variance components for responsiveness to the SICTT and abattoir confirmed M. bovis infection. The link functions between the observed 0/1 scale and the liability scale were the complementary log-log in the GB, and logit link function in the Irish population. RESULTS AND DISCUSSION: The estimated heritability of susceptibility to TB, as judged by responsiveness to the SICTT, was 0.16 (0.012) and 0.14 (0.025) in the GB and Irish populations, respectively. For abattoir or laboratory confirmation of infection, estimates were 0.18 (0.044) and 0.18 (0.041) from the GB and the Irish populations, respectively. CONCLUSIONS: Estimates were all significantly different from zero and indicate that exploitable variation exists among GB and Irish Holstein Friesian dairy cows for resistance to TB. Epidemiological analysis suggests that factors such as variation in exposure or imperfect sensitivity and specificity would have resulted in underestimation of the true values. PMID- 21645295 TI - Genome-wide association studies and genetic architecture of common human diseases. AB - Genome-wide association scans provide the first successful method to identify genetic variation contributing to risk for common complex disease. Progress in identifying genes associated with melanoma show complex relationships between genes for pigmentation and the development of melanoma. Novel risk loci account for only a small fraction of the genetic variation contributing to this and many other diseases. Large meta-analyses find additional variants, but there is current debate about the contribution of common polymorphisms, rare polymorphisms or mutations to disease risk. PMID- 21645296 TI - Genome wide scan for somatic cell counts in holstein bulls. AB - BACKGROUND: Mastitis is the most costly disease for dairy production, and control of the disease is often difficult, due to its multi-factorial nature. Susceptibility to mastitis is under partial genetic control and the industry uses indirect selection for decreased concentrations of somatic cells in milk to reduce mastitis. METHODS: A genome-wide scan was performed to identify genomic regions associated with deregressed estimated breeding values (EBVs) for somatic cell counts (SCC) in Holstein bulls. In total 1183 proven bulls of the Italian of Holstein population, were genotyped with the BovineSNP50 BeadChip (Illumina, San Diego, CA) and a whole genome association analysis was performed using the R package GenABEL. RESULTS: Two chromosomal regions showed association with SCC, a region on chromosome 14 with high significance (P < 5x10-6) and a region on chromosome 6 with moderate significance (P < 5x10-5). CONCLUSIONS: Two regions with effects on SCC have been identified with good statistical support. A further study of these candidate regions will be performed to verify the results and identify the causal mutations. PMID- 21645297 TI - Across-line SNP association study for (innate) immune and behavioral traits in laying hens. AB - BACKGROUND: An association study between single nucleotide polymorphism markers (SNP) and (innate and adaptive) immune parameters but also feather condition score on the back, rump and belly of laying hens was performed. The immune parameters measured in blood samples were natural and acquired antibody titers and complement activity. Feather condition score as a measure of feather damage was determined, this parameter is closely related to feather pecking behavior in hens housed in groups.The aim of the study was to detect associations between genetic markers and immune parameters and feather condition score across nine lines of laying hens, focusing on the feather peckers as well as on the victims of feather pecking. METHODS: A novel approach based on across-line analysis and testing of the SNP-by-line interaction was performed. RESULTS: In total 59 significant associations between SNP and immune traits were detected. Previously identified QTL were confirmed and new associations of genes regulating immune function identified. The IL17A gene (chromosome 3) influences natural and acquired antibody titers and activation of classical and alternative complement pathways. The major histocompatibility complex on chromosome 16 showed significant association with natural and acquired antibody titers and classical complement activity. The IL12B and IRF1 genes on chromosome 13 were associated with natural antibody titers.The direct effect of the genotype of an individual on its feather condition and the associative effect of the genotype of the cage mates on the individual's feather condition were analyzed. The direct genetic effect can be described as the susceptibility to be pecked at, and the associative genetic effect as the propensity to perform feather pecking. Eleven significant associations were detected for the direct effect, and 81 for the associative effect. The serotonin receptor 2C (HTR2C) on chromosome 4 was highlighted in both analyses. CONCLUSIONS: Our results confirmed previously identified QTL and identified new associations of genes regulating immune function. The results for feather condition score supports existing evidence of involvement of the serotonergic system in feather pecking in laying hens. Immune regulatory genes were found to be associated to feather condition score, revealing relationships between the immune system and behavior. PMID- 21645298 TI - The three-way relationship of polymorphisms of porcine genes encoding terminal complement components, their differential expression, and health-related phenotypes. AB - BACKGROUND: The complement system is an evolutionary ancient mechanism that plays an essential role in innate immunity and contributes to the acquired immune response. Three modes of activation, known as classical, alternative and lectin pathway, lead to the initiation of a common terminal lytic pathway. The terminal complement components (TCCs: C6, C7, C8A, C8B, and C9) are encoded by the genes C6, C7, C8A, C8B, C8G, and C9. We aimed at experimentally testing the porcine genes encoding TCCs as candidate genes for immune competence and disease resistance by addressing the three-way relationship of genotype, health related phenotype, and mRNA expression. RESULTS: Comparative sequencing of cDNAs of animals of the breeds German Landrace, Pietrain, Hampshire, Duroc, Vietnamese Potbelly Pig, and Berlin Miniature Pig (BMP) revealed 30 SNPs (21 in protein domains, 12 with AA exchange). The promoter regions (each ~1.5 kb upstream the transcription start sites) of C6, C7, C8A, C8G, and C9 exhibited 29 SNPs. Significant effects of the TCC encoding genes on hemolytic complement activity were shown in a cross of Duroc and BMP after vaccination against Mycoplasma hyopneumoniae, Aujeszky disease virus and PRRSV by analysis of variance using repeated measures mixed models. Family based association tests (FBAT) confirmed the associations. The promoter SNPs were associated with the relative abundance of TCC transcripts obtained by real time RT-PCR of 311 liver samples of commercial slaughter pigs. Complement gene expression showed significant relationship with the prevalence of acute and chronic lung lesions. CONCLUSIONS: The analyses point to considerable variation of the porcine TCC genes and promote the genes as candidate genes for disease resistance. PMID- 21645299 TI - Emerging roles of chicken and viral microRNAs in avian disease. AB - BACKGROUND: MicroRNAs are short RNAs (~22 nt) expressed by plants, animals and viruses that regulate gene expression post-transcriptionally, and their importance is highlighted by distinct patterns of expression in many physiological processes, including development, hematopoeisis, stress resistance, and disease. Our group has characterized the microRNAs encoded by the avian herpesviruses; namely, oncogenic Marek's disease (MD) virus (MDV1), non-oncogenic MDV (MDV2) herpesvirus of turkeys (HVT), and infectious laryngotracheitis virus (ILTV). METHODS: MicroRNAs encoded by the avian herpesviruses were identified using next generation sequencing technologies (454, Illumina). RESULTS: The microRNAs of each the avian herpesviruses have unique sequences, but the genomic locations are similar, in that the microRNAs tend to be clustered in the rapidly evolving repeat regions of the viral genomes. For a given viral species the microRNA sequence is highly conserved in different strains with the exception of a virulence-associated polymorphism in the putative promoter of the MDV1 microRNAs upstream of the meq oncogene. These microRNAs are relatively highly expressed in tumors produced by very virulent MDV1 isolates compared to tumors produced by less virulent strains. MDV1 and HVT encode homologs of the host microRNA, miR-221, which targets a gene important in cell cycle regulation. MDV1 encodes a microRNA (mdv1-miR-M4) that shares a seed sequence with miR-155, a microRNA important in immune function. Mdv-miR-M4 is highly expressed in MDV induced tumors, while miR-155 is present at very low levels. CONCLUSIONS: MicroRNAs are highly conserved among different field strains of MDV1, and they are expressed in lytic and latent infections and in MDV1-derived tumors. This suggests that these small molecules are very important to the virus, and roles in immune evasion, anti-apoptosis, or proliferation are likely. PMID- 21645300 TI - Preliminary association analysis of TLR9 gene polymorphisms and immune parameters in an Italian Holstein calves population. AB - BACKGROUND: This preliminary study was aimed at evaluating the association between single nucleotide polymorphisms (SNPs) on Toll like receptor 9 (TLR9) gene and some immunological parameters in a population of Italian Holstein calves. METHODS: The study was carried out in a commercial farm on 68 Holstein calves aging about 6 months. Genomic DNA was extracted from peripheral blood mononuclear cells (PBMC) and genotyped for nine SNPs on TLR9. Immunological parameters considered were the immunoglobulin (Ig) G titers against bovine herpesvirus 1, and the proliferative response of peripheral blood mononuclear cells to mitogens. For the association study, only results relative to the SNP located in the promoter region have been discussed. RESULTS: Among the nine SNPs expected, only eight were detected. Considering the SNP located in the promoter region, all three possible genotypes were observed, and their distribution was as follows: genotype a (n=34), b (n=19), and c (n=8). On the basis of their response to vaccine, calves were categorized as low (L, n=8), medium (M, n=45) and high responders (H, n=8). Although no significant association was found between genotypes and L, M or H categories, the genotype estimated as the less represented within the population (c) had no calves categorized as H, the highest frequency of L (25%), and mean values of IgG lower (P < 0.005) compared to genotype b. Furthermore, IgG titers were positively correlated with responses of PBMC to mitogens. CONCLUSIONS: Genotype c appeared to be "non advantageous" in terms of immune response. It was characterized by the presence of the mutation in homozygosity and, not surprisingly, it was the most rare genotype in the population. Larger studies are necessary in order to confirm these observations. PMID- 21645301 TI - Identification of parental line specific effects of MLF2 on resistance to coccidiosis in chickens. AB - BACKGROUND: MLF2 was the candidate gene associated with coccidiosis resistance in chickens. Although single marker analysis supported the association between MLF2 and coccidiosis resistance, causative mutation relevant to coccidiosis was not identified yet. Thus, this study suggested segregation analysis of MLF2 haplotype and the association test of the other candidate genes using improved data transformation. RESULTS: A haplotype probably originated from one parental line was found out of 4 major haplotypes of MLF2. Frequency of this haplotype was 0.2 in parental chickens and its offspring in 12 families. Allele substitution effect of the MLF2 haplotype originated from a specific line was associated with increased body weight and fecal egg count explaining coccidiosis resistance. Nevertheless Box-Cox transformation was able to improve normality; association test did not produce obvious different results compared with analysis with log transformed phenotype. CONCLUSION: Allele substitution effect analysis and classification of MLF2 haplotype identified the segregation of haplotype associated with coccidiosis resistance. The haplotype originated from a specific parental line was associated with improving disease resistance. Estimating effect of MLF2 haplotype on coccidiosis resistance will provide useful information for selecting animals or lines for future study. PMID- 21645302 TI - Whole genome analysis of infectious bovine keratoconjunctivitis in Angus cattle using Bayesian threshold models. AB - Infectious bovine keratoconjunctivitis (IBK), also known as pinkeye, is characterized by damage to the cornea and is an economically important, lowly heritable, categorical disease trait in beef cattle. Scores of eye damage were collected at weaning on 858 Angus cattle. SNP genotypes for each animal were obtained from BovineSNP50 Infinium-beadchips. Simultaneous associations of all SNP with IBK phenotype were determined using Bayes-C that treats SNP effects as random with equal variance for an assumed fraction (pi=0.999) of SNP having no effect on IBK scores. Bayes-C threshold models were used to estimate SNP effects by classifying IBK into two, three or nine ordered categories. Magnitudes of genetic variances estimated in localized regions across the genome indicated that SNP within the most informative regions accounted for much of the genetic variance of IBK and pointed out some degree of association to IBK. There are many candidate genes in these regions which could include a gene or group of genes associated with bacterial disease in cattle. PMID- 21645303 TI - Expression of trypanotolerance in N'Dama x Boran crosses under field challenge in relation to N'Dama genome content. AB - BACKGROUND: Animal trypanosomosis in sub-Saharan Africa is a major obstacle to livestock based agriculture. Control relies on drugs with increasing incidence of multiple-drug resistance. A previous mapping experiment in an F2 population derived from the indigenous trypanotolerant N'Dama cattle crossed to susceptible (Kenya)-Boran cattle under controlled challenge, uncovered a number of trypanotolerance QTL (T-QTL). The present study was to determine expression of N'Dama trypanotolerance in a backcross to the Boran under conditions of field challenge, and whether chromosomal regions associated with trypanotolerance in the F2 experiment showed similar effects in the BC population. METHODS: 192 backcross animals to the Boran were produced in six batches from June 2001 to December 2006. At one year of age animals were moved to the field and exposed to natural challenge over about one year in Southwest Kenya (Narok). The animals were individually recorded weekly for body weight, packed cell volume, parasitaemia score, and drug treatments, and were genotyped using 35 microsatellite markers spanning 5 chromosomes found in the F2 study to harbour T QTL. RESULTS: The F1 were most trypanotolerant, Boran least, and BC intermediate. Females showed distinctly higher trypanotolerance than males. There was a positive correlation in the BC population between trypanotolerance and number of N'Dama origin marker alleles. QTL mapping revealed T-QTL distributed among all five targeted chromosomes, corresponding in part to the results obtained in the F2 experiment. CONCLUSIONS: N'Dama origin trypanotolerance is expressed in a BC population under field conditions in proportion to N'Dama origin marker alleles. Consequently, marker assisted selection in such populations may be a means of increasing trypanotolerance, while retaining the desirable productive qualities of the recurrent parent. PMID- 21645304 TI - Genomic selection for carrier-state resistance in chicken commercial lines. AB - BACKGROUND: Salmonella propagation by apparently healthy chicken and subsequent food security concerns could be decreased by the selection and use of chicken lines more resistant to carrier-state. In the present study we applied the first steps of the genomic selection methodology to assess the interest of including genetic markers for the genetic evaluation of hen lines infected with Salmonella Enteritidis. METHODS: We studied commercial laying hen lines divergently selected for resistance to Salmonella carrier-state at two different ages. A total of 600 animals were typed with 1536 SNP markers and artificially infected with S. Enteritidis. Phenotypes were collected four weeks (young animals) or five weeks (adults) later. Two types of variance component analyses, including or not including SNP data, were performed and compared. All variance components were estimated by Bayesian methods and Gibbs sampling. RESULTS: The comparison of both genetic analyses shows that SNP are efficient in capturing genetic variation, although none of them captures a large affect on the traits studied. Average accuracies do not change between analyses, showing that using SNP data does not really increase information. CONCLUSIONS: These preliminary results show that genomic selection for Salmonella carrier-state resistance in laying hens is promising, although a denser SNP coverage of the genome on a higher number of animals is needed to assess its feasibility and efficiency compared to classical pedigree evaluation. PMID- 21645305 TI - Genomic study of the response of chicken to highly pathogenic avian influenza virus. AB - BACKGROUND: The host mounts an immune response to pathogens, but few data are currently available on the role of host genetics in variation in response to avian influenza (AI). The study presented here investigated the role of the host genetic background in response to in vivo infection with AI virus (AIV). METHODS: Experimental lines of chicken and commercial crosses were experimentally infected intratracheally with 103 EID50/bird of A/Chicken/Italy/13474/99 H7N1 highly pathogenic avian influenza virus (HPAIV). Chickens were genotyped for the Mx polymorphism causing the S631N mutation, and for the Major Histocompatibility Complex (MHC). Whole-genome genotyping was carried out using 60 k Single Nucleotide Polymorphism (SNP) array developed by the poultry Genome-Wide Marker Assisted Selection Consortium (GWMASC). RESULTS: Variability in response of different chicken lines to the HPAIV infections and some degree of resistance to AI were observed: a statistically significant effect of chicken line on the response to infection was found. There was no association between survival in healthy conditions and polymorphisms at the Mx gene and the MHC-B region. The analysis based on the 60 k SNPs provided a good clustering of the chicken lines, but no specific genetic cluster associated with response to AIV was identified. CONCLUSIONS: Neither the genotype at the Mx gene or MHC-B locus, nor for SNP spanning the whole-genome identified loci involved in variations to response to AIV infection. These results point towards the possibility that either the genetic factors affecting the response of chickens to the H7N1 HPAIV are weak, or relevant alleles were not segregating in the studied populations. PMID- 21645306 TI - Insights into gene expression profiling of natural resistance to coccidiosis in contrasting chicken lines. AB - Coccidiosis is a parasitic disease with major economic impact, one of whose main causative agents is Eimeria tenella. Chicken breeds display variable natural resistance to this disease. Unravelling the genetic bases of such variations could provide new clues for protection strategies. Transcriptomic experiments were conducted comparing resistant (Fayoumi) and susceptible (Leghorn) lines. Caecum and caecal tonsils were analysed. A global increase in differential gene expression following infection was observed for caecum comparisons, whereas a global decrease following infection was observed for caecal tonsils.Gene lists for infected tissues display 40 genes in common across breeds, 20 of which were specific to infected tissues. Among these specific genes, 9 belong to the 100 more differentially expressed genes of the infected caecum comparison. Gene expression networks were constructed in parallel, identifying highly connected genes. Comparing information from differential gene lists and gene network analysis allows one to highlight potential pivotal genes in the infection process, one of which was located in a putative significant QTL region for infection associated lesions. PMID- 21645307 TI - Polymorphisms in pattern recognition receptors and their relationship to infectious disease susceptibility in pigs. AB - BACKGROUND: Pattern recognition receptors (PRRs), including Toll-like receptors (TLRs), are censoring receptors for molecules derived from bacteria, viruses, and fungi. The PRR system is a prerequisite for proper responses to pathogens, for example by cytokine production, resulting in pathogen eradication. Many cases of polymorphisms in PRR genes affecting the immune response and disease susceptibility are known in humans and mice. METHODS: We surveyed polymorphisms in pig genes encoding PRRs and investigated the relationship between some of the detected polymorphisms and molecular function or disease onset. RESULTS: Nonsynonymous polymorphisms abounded in pig TLR genes, particularly in the region corresponding to the ectodomains of TLRs expressed on the cell surface. Intracellular TLRs such as TLR3, TLR7, and TLR8, and other intracellular PRRs, such as the peptidoglycan receptor NOD2 and viral RNA receptors RIG-I and MDA5, also possessed nonsynonymous polymorphisms. Several of the polymorphisms influenced molecular functions such as ligand recognition. Polymorphisms in the PRR genes may be related to disease susceptibility in pigs: pigs with a particular allele of TLR2 showed an increased tendency to contract pneumonia. CONCLUSIONS: We propose the possibility of pig breeding aimed at disease resistance by the selection of PRR gene alleles that affect pathogen recognition. PMID- 21645308 TI - Using SNP array data to test for host genetic and breed effects on Porcine Reproductive and Respiratory Syndrome Viremia. AB - BACKGROUND: The effect of breed on Porcine Reproductive and Respiratory Syndrome Viremia (PRRSV) was tested using data collected in 17 Italian commercial pig farms and 1096 genotypes obtained by the PorcineSNP60 BeadChip. A binomial logistic model was used to investigate the relationship between breed-clusters and PRRSV susceptibility. Breed-clusters were defined using the matrix of genomic kinship between all pairs of piglets. RESULTS: Only the contemporary group effect, defined as all piglets reared in the same herd, in the same year and whose samples were collected in the same season, was significant. Sex, age and breed-cluster showed no statistically significant effect on PRRS viremia, although the Landrace and Cross breed-clusters showed the lowest Odds-Ratio CONCLUSIONS: The model failed to detect a significant breed-cluster effect, highlighting the impact of environment and management on PRRS viremia incidence. Incomplete exposure over the observed period may have masked possible breed differences. PMID- 21645309 TI - Correlation in chicken between the marker LEI0258 alleles and Major Histocompatibility Complex sequences. AB - BACKGROUND: The LEI0258 marker is located within the B region of the chicken Major Histocompatibility Complex (MHC), and is surprisingly well associated with serology. Therefore, the correlation between the LEI0258 alleles and the MHC class I and the class II alleles at the level of sequences is worth investigating in chickens. Here we describe to which extent the LEI0258 alleles are associated with alleles of classical class I genes and non-classical class II genes, in reference animals as well as local breeds with unknown MHC haplotypes. METHODS: For the class I region, in an exploratory project, we studied 10 animals from 3 breeds: Rhode Island Red, White Leghorn and Fayoumi chickens, by cloning and sequencing B-F1 and B-F2 cDNA from exon 1 to 3'UTR. For the class II region, we reconstructed haplotypes of the 8.8 kb genomic region encompassing three non classical class II genes: B-DMA, B-DMB1 and B-DMB2, for 146 animals from more than 50 breeds including wild species of jungle fowls. RESULTS: Overall we found that the LEI0258 marker genotypes gave good indications of the MHC haplotypes, and a very good predictions (>0.95) of the heterozygosity of an animal at the MHC locus. CONCLUSIONS: Our results show that the LEI0258 alleles are strongly associated with haplotypes of classical class I genes and non-classical class II genes, unravelling the reasons why this marker is becoming the reference marker for MHC genotyping in chickens. PMID- 21645310 TI - Comparison of DNA methylation levels of repetitive loci during bovine development. AB - BACKGROUND: DNA methylation of cytosine residues in CpG dinucleotide controls gene expression and dramatically changes during development. Its pattern is disrupted in cloned animals suggesting incomplete reprogramming during somatic cell nuclear transfer (the first reprogramming). However, the second reprogramming occurs in the germ cells and epigenetic errors in somatic cells of cloned animals should be erased. To analyze the DNA methylation changes on the spermatogenesis of bulls, we measured DNA methylation levels of three repetitive elements in blastocysts, blood and sperm. METHODS: DNA from PBLs (peripheral blood leukocytes), sperm and individual IVF (in vitro fertilized) and parthenogenetic blastocysts was isolated and bisulfite converted. Three repetitive elements; Satellite I, Satellite II and art2 sequences were amplified by PCR with specific pairs of primers. The PCR product was then cut by restriction enzymes and analyzed by agarose gel electrophoresis for determining the DNA methylation levels. RESULTS: Both Satellite I and Satellite II sequences were highly methylated in PBLs, whereas hypo-methylated in sperm and blastocysts. The art2 sequence was half methylated both in PBLs and sperm but less methylated in blastocysts. There was no difference in DNA methylation levels between IVF and parthenogenetic blastocysts. CONCLUSIONS: These results suggest that there is a dynamic change of DNA methylation during embryonic development and spermatogenesis in cattle. Satellite I and Satellite II regions are methylated during embryogenesis and then de-methylated during spermatogenesis. However, art2 sequences are not de-methylated during spermatogenesis, suggesting that this region is not reprogrammed during germ cell development. These results show dynamic changes of DNA methylation levels during bovine embryogenesis, especially genome-wide reprogramming in germ cells. PMID- 21645311 TI - Probing genetic control of swine responses to PRRSV infection: current progress of the PRRS host genetics consortium. AB - BACKGROUND: Understanding the role of host genetics in resistance to porcine reproductive and respiratory syndrome virus (PRRSV) infection, and the effects of PRRS on pig health and related growth, are goals of the PRRS Host Genetics Consortium (PHGC). METHODS: The project uses a nursery pig model to assess pig resistance/susceptibility to primary PRRSV infection. To date, 6 groups of 200 crossbred pigs from high health farms were donated by commercial sources. After acclimation, the pigs were infected with PRRSV in a biosecure facility and followed for 42 days post infection (dpi). Blood samples were collected at 0, 4, 7, 10, 14, 21, 28, 35 and 42 dpi for serum and whole blood RNA gene expression analyses; weekly weights were recorded for growth traits. All data have been entered into the PHGC relational database. Genomic DNAs from all PHGC1-6 pigs were prepared and genotyped with the Porcine SNP60 SNPchip. RESULTS: Results have affirmed that all challenged pigs become PRRSV infected with peak viremia being observed between 4-21 dpi. Multivariate statistical analyses of viral load and weight data have identified PHGC pigs in different virus/weight categories. Sera are now being compared for factors involved in recovery from infection, including speed of response and levels of immune cytokines. Genome-wide association studies (GWAS) are underway to identify genes and chromosomal locations that identify PRRS resistant/susceptible pigs and pigs able to maintain growth while infected with PRRSV. CONCLUSIONS: Overall, the PHGC project will enable researchers to discover and verify important genotypes and phenotypes that predict resistance/susceptibility to PRRSV infection. The availability of PHGC samples provides a unique opportunity to continue to develop deeper phenotypes on every PRRSV infected pig. PMID- 21645312 TI - Genetic and cellular aspects of the establishment of histocompatible stem cells: information gained from an animal model. AB - The establishment of patient-specific histocompatible stem cells may be an alternative for overcoming current limitations in stem cell engineering. We are developing an animal model to assist the establishment of histocompatible, autologous stem cells. In this process, we obtained valuable information on establishing and characterizing stem cells. As an initial step, we succeeded in establishing histocompatible stem cells using preantral follicle cultures and subsequent parthenogenetic activation. The gene expression profile of the established stem cells was similar to that of embryonic stem cells (ESCs) derived from normal fertilization. On the other hand, we propose a way to derive histocompatible, ESC-like cells by co-culturing ovarian stromal cells with feeder fibroblasts, which may allow the derivation of stem cells from somatic tissue. However, more progress regarding the establishment and elucidation on origination of established cell lines is necessary to use this genetic manipulation-free procedure. Nevertheless, relevant information on the process will help to stimulate preclinical research on cell transformation into differentiated, undifferentiated, and even cancerous cells, as well as clinical studies on the application of induced pluripotent cells. PMID- 21645313 TI - Deciphering the genetic control of innate and adaptive immune responses in pig: a combined genetic and genomic study. AB - Improving animal robustness and resistance to pathogens by adding health criteria in selection schemes is one of the challenging objectives of the next decade. In order to better understand the genetic control of immunity in French Large White pigs, we have launched a program combining genetic and genomic studies not focussing on any particular pathogen. Animals recorded for production traits were scored for a wide range of immunity parameters three weeks after vaccination against Mycoplasma hyopneumoniae: i) total white blood cells and lymphocyte counts and proportions of various leucocyte subsets including cells harbouring IgM, gammadeltaTCR, CD4/CD8, CD16/CD2 and CD16/CD172a/MHCII, ii) innate immune response parameters (phagocytosis and in vitro production of IL1B, IL6, IL8, TNF, IL12 and IFNalphaafter blood stimulation), iii) adaptive immune response parameters (lymphocyte proliferation, in vitro production of IL2, IL4, IL10 and IFNgamma after blood stimulation, total IgG, IgA, IgM and specific IgG levels) and iv) two acute phase proteins (C-reactive protein and haploglobin). Across traits, heritability estimates reached 0.4 on average (se=0.1) and 42 of the 54 measured parameters showed moderate to high heritabilities (>=0.2), confirming that many parameters are under genetic control and could be included in selection protocols. Functional analyses revealed that the blood transcriptome is informative for part of the immunity traits and should provide relevant phenotypic information to better characterize some immunity traits. PMID- 21645314 TI - Different immune responses to three different vaccines following H6N1 low pathogenic avian influenza virus challenge in Taiwanese local chicken breeds. AB - BACKGROUND: H6N1 low pathogenic avian influenza virus (LPAIV) are frequently isolated in Taiwan and lead to significant economic losses, either directly or indirectly through association with other infectious diseases. This study investigates immune responses to three different vaccines following a H6N1 challenge in different local breeds. METHODS: Experimental animals were sampled from six local chicken breeds maintained at the National Chung-Hsing University, namely Hsin-Yi, Ju-Chi, Hua-Tung (Taiwan), Quemoy (Quemoy Island), Shek-Ki (China), Nagoya (Japan) and a specific pathogen free (SPF) White Leghorn line. A total number of 338 chickens have been distributed between a control and a challenge group, H6N1 challenge was performed at 7 weeks of age; vaccination against Newcastle Disease (ND), Infectious Bursal Disease (IBD) and Infectious Bronchitis (IB) was performed at 11 weeks. The anti-H6N1 LPAIV antibody titers were measured by ELISA at days 0, 7, 14 and 21 after challenge, and the anti-ND, anti-IBD and anti-IB antibody titers were measured by inhibition of hemagglutination test and ELISA at days 0, 14, 28 after vaccination. RESULTS: There was no effect of the H6N1 LPAIV challenge at 7 weeks of age on the subsequent responses to ND and IBD vaccine at 11 weeks of age, but, surprisingly, the H6N1 LPAIV challenge significantly affected antibody levels to IB vaccine in some breeds, since IB0 and IB14 antibody titers were lower in the challenge groups. However, there was no significant difference in IB28 antibody titers among the experimental groups. CONCLUSIONS: Local breeds have different immune response to H6N1 LPAIV challenge and subsequent vaccines. Differences dealt mainly with kinetics of response and with peak values. Quemoy exhibited higher antibody levels to H6N1, ND and IBD. The negative effect of the H6N1 LPAIV challenge on IB vaccine response may be related to the fact that both viruses target the lung tissues, and the type of local immune response induced by LPAIV challenge may not be favourable for birds to make optimum IB-specific antibody response. PMID- 21645315 TI - Effects of dietary plant-derived phytonutrients on the genome-wide profiles and coccidiosis resistance in the broiler chickens. AB - BACKGROUND: The present study was conducted to investigate the effects of dietary plant-derived phytonutrients, carvacrol, cinnamaldehyde and Capsicum oleoresin, on the translational regulation of genes associated with immunology, physiology and metabolism using high-throughput microarray analysis and in vivo disease challenge model of avian coccidiosis. METHODS: In this study, we used nutrigenomics technology to investigate the molecular and genetic mechanisms of dietary modulation of host innate immunity and metabolism by three phytonutrients. To validate their immunomodulatory effects in a disease model, young broiler chickens fed a standard diet supplemented with three phytochemicals (carvacrol, cinnamaldehyde, and Capsicum oleoresin) from one day post-hatch were orally challenged with E. acervulina. The body weight gain and fecal oocyst production were used to evaluate coccidiosis disease parameters. RESULTS: Analysis of global gene expression profiles of intestinal tissues from phytonutrient-fed birds indicated that Capsicum oleoresin induced the most gene changes compared to the control group where many of these genes were associated with those of metabolism and immunity. The most reliable network induced by dietary cinnamaldehyde treatment was related with the functions of antigen presentation, humoral immune response, and inflammatory disease. Furthermore, dietary supplementation with these phytonutrients significantly protected broiler chickens against live coccidiosis challenge infection based on body weight and parasite fecundity. CONCLUSIONS: The results of this study provide clear evidence to support the idea that plant-derived phytochemicals possess immune-enhancing properties in chickens and these new findings create a new possibility to develop effective drug-free alternative strategies for disease control for poultry infectious diseases. PMID- 21645316 TI - Dietary unsaturated fatty acids affect the mammary gland integrity and health in lactating dairy cows. AB - BACKGROUND: Information about the effects of unsaturated fatty acids (UFA) supplementation on the health and integrity of the mammary gland in lactating dairy cows is lacking. Therefore, the aim of this study was to determine the effects of unprotected dietary UFA on the global expression pattern of genes in the mammary gland tissue of grazing dairy cows, and to translate this information into relevant biological knowledge. METHODS: Twenty-eight Holstein-Friesian dairy cows were randomly assigned to 4 different concentrated UFA-sources for 23 days after which all cows were switched to a non-UFA-supplemented concentrate for an additional 28 days. On the last day of both periods, mammary gland biopsies were taken to study genome-wide differences in gene expression on Bovine Genome Arrays. RESULTS: Supplementation with UFA reduced the concentration of short chain fatty acids (FA), C16 FA and saturated FA in the milk, whereas that of trans-FA increased. One major finding was that canonical pathways associated with remodelling and immune functions of the mammary gland were predominantly down regulated during UFA supplementation and negatively correlated with the concentration of milk trans-FA. CONCLUSIONS: Supplementing grazing dairy cows with unprotected dietary UFA can affect the remodelling and immune functions of the mammary gland with potential consequences for its integrity and health, as well as milk quality. PMID- 21645317 TI - In silico analysis of candidate genes associated with humoral innate immune response in chicken. AB - BACKGROUND: Production and function of natural antibodies (NAbs) constitutes an important mechanism of the humoral innate immunity in vertebrates. The level of NAbs in chicken is heritable and the genetic background has been partly investigated. However, to date the genetic determination of humoral innate immune response in avian species has not been fully described. The goal of this study was to propose a new set of candidate genes with a potential effect on the NAb phenotype for further SNP association study. METHODS: In silico analysis of positional and functional candidate genes covered 14 QTL regions associated with LPS, LTA & KLH NAbs and located on six chromosomes: GGA5, GGA6, GGA9, GGA14, GGA18 and GGAZ. The function of the genes was subsequently determined based on the NCBI, KEGG, Gene Ontology and InnateDB databases. RESULTS: As a result, the core panel of 38 genes participating in metabolic pathways of innate immune response was proposed. Most of them were assigned to chromosomes: GGA14, GGA5, GGA6 and GGAZ (13, 9, 8 and 5 genes, respectively). These candidate genes encode proteins predicted to play a role in (i) proliferation, differentiation and function of B lymphocytes; (ii) TLR signalling pathway, and (iii) MAP signalling cascade. CONCLUSIONS: Proposed set of candidate genes is recommended to be included in the follow-up studies to model genetic networks of innate humoral immune response in chicken. PMID- 21645318 TI - Selection of Mx gene genotype as genetic marker for Avian Influenza resistance in Indonesian native chicken. AB - BACKGROUND: In previous studies, the Mx Gene has been demonstrated to confer positive anti viral responses in chicken. The amino acid variation of Asn (allele A) at position 631 was specific to positive antiviral Mx/resistant, while, that of Ser (allele G) was specific to negative Mx/susceptible. This research was aimed at selecting one of the native chicken breeds which was found out to be resistant to avian influenza using molecular technique. The selected breed will then be used as the base population to improve native chicken breed in Indonesia. METHODS: Marker Assisted Selection (MAS) method was used in this research to accelerate the selection process, since the disease resistance had low heritability value. Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) technique used to select the genotype of Mx++, Mx+- and Mx-- that corresponded to the positive antiviral activity (Mx++), or those which had positive or negative activity (Mx+-) and negative antiviral activity (Mx--). There were 200 native hens and 40 cocks used in this experiment. Allele frequency of Mx Gene was calculated. The productivity indicators such as age at first laying, egg weight and hen weight at first laying and egg production were also measured. The chicken that had Mx++ and Mx+- genotypes, were selected to produce offspring. RESULTS: Result showed that the frequency of the resistant allele (Mx+) was 65% and 60% in laying hens and in cocks, respectively, while the frequency of the susceptible allele (Mx-) was 35% and 40% in hens and cocks, resepctively. Age, egg weight and hen weight at first laying and egg production for susceptible genotype were slightly better than for the resistant genotype which were 172,41 VS 178,81 days; 33,94 VS 32,84 g; 1450 VS 1439 g and 54,32 VS 48,30 %, respectively. PMID- 21645319 TI - Jejunal gene expression patterns correlate with severity of systemic infection in chicken. AB - BACKGROUND: Not much is known about the effect of Salmonella enteritidis on changes in the developmental processes occurring in the intestine of young chicken. Therefore we investigated the correlation of intestinal gene expression patterns with the severity of systemic Salmonella infections. METHODS: The number of Salmonella colony forming units (CFUs) in the liver of infected chicken were plotted against the average intestinal expression profiles of previously identified gene expression clusters. The functional properties of all the genes taken together present in 3 clusters exhibiting positive correlation at early time-points were compared with the functional properties of the genes displaying antagonistic correlations in 1 cluster. The top 5 ranking functional groups were analysed in further detail. RESULTS: Three clusters showed gene expression profiles which were positively correlated with the severity of systemic disease as measured by the number of Salmonella colony forming units in the liver. In these clusters, genes involved in morphological processes were predominantly present. One cluster had a profile that was negatively correlated with the severity of systemic disease, as measured by numbers of CFUs in the liver. The genes in the latter cluster were mostly involved in cell turn-over and metabolism. CONCLUSIONS: In the developing jejunum of young chicken, both stimulatory and inhibitory gene expression mechanisms are correlated with the severity of systemic Salmonella infections. PMID- 21645320 TI - Contribution of the genetic background to the immune response of broilers vaccinated or challenged with LPAI H9N2. AB - BACKGROUND: The knowledge on the immune responses to LPAI is limited. The purpose of this study was to investigate the immune responses of two divergently selected lines of broilers, a line responding with high antibody response to antigens (HH), and a line responding with low antibody titers (LL) to antigen. METHODS: Day old chicks from each line were divided in two groups, one vaccinated with inactivated H9N2 vaccine and one non-vaccinated. At 21 days of age all the chicks were challenged with field isolate of H9N2, 1X106.5 ELD50 per chick by drops to the eye, nose and beak. Twenty four hours and 14 days post challenge (PC), the chickens were weighed blood spleen and lungs were taken and leukocytes were isolated. The leukocytes were stained with monoclonal antibodies for surface markers and analyzed by flow cytometry. We used Elispot assay to identify the number of antibody producing cells in each of the organs. mRNA was extracted using TRIsol reagent in order to assess the cytokine production level by qRT-PCR using the SYBR green methods. RESULTS: Our results showed that LL-vaccinated group gained more weight than any of the other group. Using IDEXX kit, no antibody titers could be identified in vaccinated chicks 21 days post vaccination while 14 days PC vaccinated HH chickens demonstrated the highest average antibody titers. LL vaccinated chickens demonstrated higher average antibody titer than non-vaccinated LL. Using the Elispot assay no difference were found between the groups either cells producing IgA, IgM or IgY beside of a high number of IgY producing cells in the lungs of vaccinated HH birds. CONCLUSIONS: Further data on leukocytes subpopulations using flow cytometry, cytokines production (IFNgamma, IL-6, IL-18, IL-2 and IL-4) isotype specific antibody responses and number and functionality of NK cells are in process. PMID- 21645321 TI - Multi-comparative systems biology analysis reveals time-course biosignatures of in vivo bovine pathway responses to B.melitensis, S.enterica Typhimurium and M.avium paratuberculosis. AB - BACKGROUND: To decipher the complexity and improve the understanding of host pathogen interactions, biologists must adopt new system level approaches in which the hierarchy of biological interactions and dynamics can be studied. This paper presents the application of systems biology for the cross-comparative analysis and interactome modeling of three different infectious agents, leading to the identification of novel, unique and common molecular host responses (biosignatures). METHODS: A computational systems biology method was utilized to create interactome models of the host responses to Brucella melitensis (BMEL), Salmonella enterica Typhimurium (STM) and Mycobacterium avium paratuberculosis (MAP). A bovine ligated ileal loop biological model was employed to capture the host gene expression response at four time points post infection. New methods based on Dynamic Bayesian Network (DBN) machine learning were employed to conduct a systematic comparative analysis of pathway and Gene Ontology category perturbations. RESULTS: A cross-comparative assessment of 219 pathways and 1620 gene ontology (GO) categories was performed on each pathogen-host condition. Both unique and common pathway and GO perturbations indicated remarkable temporal differences in pathogen-host response profiles. Highly discriminatory pathways were selected from each pathogen condition to create a common system level interactome model comprised of 622 genes. This model was trained with data from each pathogen condition to capture unique and common gene expression features and relationships leading to the identification of candidate host-pathogen points of interactions and discriminatory biosignatures. CONCLUSIONS: Our results provide deeper understanding of the overall complexity of host defensive and pathogen invasion processes as well as the identification of novel host-pathogen interactions. The application of advanced computational methods for developing interactome models based on DBN has proven to be instrumental in conducting multi conditional cross-comparative analyses. Further, this approach generates a fully simulateable model with capabilities for predictive analysis as well as for diagnostic pattern recognition. The resulting biosignatures may represent future targets for identification of emerging pathogens as well as for development of antimicrobial drugs, immunotherapeutics, or vaccines for prevention and treatment of diseases caused by known, emerging/re-emerging infectious agents. PMID- 21645322 TI - Down-regulation of promoter methylation level of CD4 gene after MDV infection in MD-susceptible chicken line. AB - BACKGROUND: Marek's disease virus (MDV) is an oncovirus that induces lymphoid tumors in susceptible chickens, and may affect the epigenetic stability of the CD4 gene. The purpose of this study was to find the effect of MDV infection on DNA methylation status of the CD4 gene differed between MD-resistant (L63) and susceptible (L72) chicken lines. METHODS: Chickens from each line were divided into two groups with one group infected by MDV and the other group as uninfected controls. Then, promoter DNA methylation levels of the CD4 gene were measured by Pyrosequencing; and gene expression analysis was performed by quantitative PCR. RESULTS: Promoter methylation of the CD4 gene was found to be down-regulated in L72 chickens only after MDV infection. The methylation down-regulation of the CD4 promoter is negatively correlated with up-regulation of CD4 gene expression in the L72 spleen at 21 dpi. CONCLUSIONS: The methylation fluctuation and mRNA expression change of CD4 gene induced by MDV infection suggested a unique epigenetic mechanism existed in MD-susceptible chickens. PMID- 21645323 TI - Porcine type I interferons: polymorphic sequences and activity against PRRSV. AB - BACKGROUND: Type I interferons (IFN) are a heterogeneous group of cytokines central to innate and adaptive antiviral immune responses. We have recently reported that porcine type I IFNs comprise at least 39 functional genes with diverse antiviral activity against porcine reproductive and respiratory syndrome virus (PRRSV). Here we report that potential cytokine polymorphisms exist in several genes of porcine type I IFNs. RESULTS: We have detected more than 100 potential polymorphic mutations, which include nucleotide substitutions and deletions, within the coding regions of porcine type I IFNs. Approximately 50% of the nucleotide changes were mutations that resulted in non-conserved amino acid substitution, as well as deletions that produced frame shifts in the open reading frames (ORFs). We have identified more than 20 polymorphic mutants that showed alterations in anti-PRRSV and anti-vesicular stomatitis virus (VSV) activity in vitro. In particular, some mutations in IFN-alpha2, IFN-alpha3, IFN-alpha8, IFN alpha12 and IFN-omega5 significantly altered the antiviral activity of expressed proteins in comparison to the wild-type or variant with more similarity to the wild-type. CONCLUSIONS: Multiple polymorphic isoforms potentially exist within subtypes of the porcine type I IFN family. Polymorphic mutations are more common in multiple-member subtypes than single-member subtypes, and most are found within the IFN-alpha subclass. Some polymorphic isoforms have altered amino acid composition and shifted ORFs, which show significantly different antiviral activity in vitro. PMID- 21645324 TI - Toll-Like Receptor 6 differential expression in two pig genetic groups vaccinated against Mycoplasma hyopneumoniae. AB - BACKGROUND: Mycoplasma hyopneumoniae is the etiologic agent of enzootic pneumonia, which causes important economic losses to swine industry. The Toll like receptors (TLRs) are pattern-recognition receptors which detect microbial presence and initiate the innate as well as the adaptative immune defense. Toll like receptor 6 is a type I transmembrane protein that recognizes bacterial components. The aim of this study was to compare mRNA expression pattern of TLR6 gene in two genetically distinct groups of pigs vaccinated against Mycoplasma hyopneumoniae. METHODS: For each genetic group, peripheral blood was collected just before and 10 days after vaccination from 10 Naturalized Brazilian Piau breed and 10 Commercial White Line serum-negative female piglets. RNA was extracted from peripheral blood mononuclear cells (PBMCs), reverse transcripted and the qRT-PCR performed using SYBR green fluorescence system, using GAPDH gene as endogenous control. Analyses were performed by UNIVARIATE (Shapiro-Wilk test) and MIXED procedures of SAS software (version 9.0). RESULTS: It was observed significant interaction between breed and vaccination, being the TLR6 mRNA expression higher in the Commercial White line than in the Piau breed after vaccination. Furthermore, there was differential expression before and after vaccination in the Commercial White line. CONCLUSIONS: Analysis of in TLR6 gene expression showed difference between the two distinct genetic groups, however, other TLRs gene expression must be evaluated for a better understanding of innate resistance in the pig concerning Mycoplasma hyopneumoniae infection. PMID- 21645325 TI - Energy and macronutrient intake and dietary pattern among school children in Bahrain: a cross-sectional study. AB - BACKGROUND: Obesity is increasing in Bahrain and there is lack of information on the energy and macronutrient intake of children. The objective of this research was to study the energy and macronutrient intake as well as food frequency pattern of Bahraini school children. METHODS: This is a cross-sectional descriptive study conducted on Bahraini school boys and girls aged 6-18 years from all the 11 populated regions of the country. Data on food intake consisted of a 24-hour dietary recall and was obtained by interviewing a sub-sample of the study population. Information was also obtained through a self-administered questionnaire for the entire sample on the weekly frequency of food items that were grouped into 7 categories based on similarity of nutrient profiles. Dietary analysis was performed using the Nutritionist 5 (First Data Bank Version 1.6 1998). RESULTS: While the average energy intake of students was close to the Estimated Average Requirements of the UK Reference standards, protein intake substantially exceeded the Reference Nutrient Intake values as did daily sugar consumption. Dietary fiber fell short of the Dietary Recommended Values (UK) and 36%-50% students exceeded the Energy % limits for total fat, saturated fat and cholesterol. The Polyunsaturated: Saturated fat ratio remained at an unacceptable level of 0.6 for girls and boys. While sweets, snacks and regular soda drinks were popular, milk, fruits and vegetables were not commonly consumed. CONCLUSIONS: High sugar consumption, low intake of dietary fiber and high energy % of saturated fat and dietary cholesterol by many Bahraini children, is likely to increase their risk of obesity and cardiovascular diseases in later life. Nutrition education programs in schools should emphasize the importance of healthy balanced diets for growth and health maintenance of children as well as dietary prevention of diseases. PMID- 21645326 TI - DJ-1 can inhibit microtubule associated protein 1 B formed aggregates. AB - BACKGROUND: Abnormal accumulation and aggregation of microtubule associated proteins (MAPs) plays an important role in the pathogenesis of neurodegenerative diseases. Loss-of-function mutation of DJ-1/Park7 can cause early onset of PD. DJ 1, a molecular chaperone, can inhibit alpha-synuclein aggregation. Currently, little is known whether or not loss of function of DJ-1 contributes to abnormal MAPs aggregation in neurodegenerative disorders such as PD. RESULTS: We presented evidence that DJ-1 could bind to microtubule associated protein1b Light Chain (MAP1b-LC). Overexpression of DJ-1 prevented MAP1b-LC aggregation in HEK293t and SH-SY5Y cells while DJ-1 knocking down (KD) enhanced MAP1b-LC aggregation in SH SY5Y cells. The increase in insoluble MAP1b-LC was also observed in the DJ-1 null mice brain. Moreover, in the DJ-1 KD SH-SY5Y cells, overexpression of MAP1B-LC led to endoplasmic reticulum (ER) stress-induced apoptosis. CONCLUSION: Our results suggest that DJ-1 acts as a molecular chaperone to inhibit MAP1B aggregation thus leading to neuronal apoptosis. Our study provides a novel insight into the mechanisms that underly the pathogenesis of Parkinson's disease (PD). PMID- 21645327 TI - Gene expression in bryozoan larvae suggest a fundamental importance of pre patterned blastemic cells in the bryozoan life-cycle. AB - BACKGROUND: Bryozoa is a clade of aquatic protostomes. The bryozoan life cycle typically comprises a larval stage, which metamorphoses into a sessile adult that proliferates by asexual budding to form colonies. The homology of bryozoan larvae with other protostome larvae is enigmatic. Bryozoan larvae exhibit blastemic tissues that contribute to build the adult during morphogenesis. However, it remains unclear if the cells of these tissues are pre-determined according to their future fate or if the cells are undifferentiated, pluripotent stem cells. Gene expression studies can help to identify molecular patterning of larval and adult tissues and enlighten the evolution of bryozoan life cycle stages. RESULTS: We investigated the spatial expression of 13 developmental genes in the larval stage of the gymnolaemate bryozoan Bugula neritina. We found most genes expressed in discrete regions in larval blastemic tissues that form definitive components of the adult body plan. Only two of the 13 genes, BnTropomyosin and BnFoxAB, were exclusively expressed in larval tissues that are discarded during metamorphosis. CONCLUSIONS: Our results suggest that the larval blastemas in Bugula are pre patterned according to their future fate in the adult. The gene expression patterns indicate that some of the bryozoan blastemas can be interpreted to correspond to homologous adult tissues of other animals. This study challenges an earlier proposed view that metazoan larvae share homologous undifferentiated "set aside cells", and instead points to an independent origin of the bryozoan larval stage with respect to other lophotrochozoans. PMID- 21645328 TI - A survey of Italian and Spanish neonatologists and paediatricians regarding awareness of the diagnosis of FAS and FASD and maternal ethanol use during pregnancy. AB - BACKGROUND: Ethanol is the most widely used drug in the world and a human teratogen whose consumption among women of childbearing age has been steadily increasing. There are no Italian or Spanish statistics on ethanol consumption during pregnancy nor any information regarding prevalence of fetal alcohol syndrome (FAS) and fetal alcohol spectrum disorders (FASD). There is also a reasonable suspicion that these two diseases are underdiagnosed by professionals from the above-reported countries. The objectives of this study were: 1) to evaluate the experience, knowledge and confidence of Italian and Spanish neonatologists and paediatricians with respect to the diagnosis of FAS and FASD, and 2) to evaluate professionals awareness of maternal drinking patterns during pregnancy. METHODS: A multiple-choice anonymous questionnaire was e-mailed to Italian neonatologists registered in the mailing list of the corresponding Society and administered to Italian and Spanish paediatricians during their National Congress. RESULTS: The response rate was 16% (63/400) for the Italian neonatologists of the National Society while a total of 152 Spanish and 41 Italian paediatricians agreed to complete the questionnaire during National Congress. Over 90% of the surveyed physicians declared that FAS is an identifiable syndrome and over 60% of them identified at least one of the most important features of FAS. Although over 60% Italian responders and around 80% Spanish responders were aware that ethanol use in pregnancy is dangerous, approximately 50% Italian responders and 40% Spanish ones allowed women to drink sometimes a glass of wine or beer during pregnancy.Neonatologists and paediatricians rated confidence in the ability to diagnosis FAS and FASD as low, with over 50% responders feeling they needed more information regarding FAS and FASD identification in newborn and child. CONCLUSIONS: Italian and Spanish neonatologists and paediatricians do not feel confident about diagnosing FAS and FASD. More training is needed in order to accurately diagnose ethanol use during pregnancy and correctly inform pregnant women on the consequences on the newborn. PMID- 21645329 TI - Transcription factors Sp1 and Sp4 regulate TRPV1 gene expression in rat sensory neurons. AB - BACKGROUND: The capsaicin receptor, transient receptor potential vanilloid type 1 (TRPV1) directs complex roles in signal transduction including the detection of noxious stimuli arising from cellular injury and inflammation. Under pathophysiologic conditions, TRPV1 mRNA and receptor protein expression are elevated in dorsal root ganglion (DRG) neurons for weeks to months and is associated with hyperalgesia. Building on our previous isolation of a promoter system for the rat TRPV1 gene, we investigated the proximal TRPV1 P2-promoter by first identifying candidate Sp1-like transcription factors bound in vivo to the P2-promoter using chromatin immunoprecipitation (ChIP) assay. We then performed deletion analysis of GC-box binding sites, and quantified promoter activity under conditions of Sp1 / Sp4 over-expression versus inhibition/knockdown. mRNA encoding Sp1, Sp4 and TRPV1 were quantified by qRT-PCR under conditions of Sp1/Sp4 over-expression or siRNA mediated knockdown in cultured DRG neurons. RESULTS: Using ChIP analysis of DRG tissue, we demonstrated that Sp1 and Sp4 are bound to the candidate GC-box site region within the endogenous TRPV1 P2 promoter. Deletion of GC-box "a" or "a + b" within the P2- promoter resulted in a complete loss of transcriptional activity indicating that GC-box "a" was the critical site for promoter activation. Co-transfection of Sp1 increased P2 promoter activity in cultured DRG neurons whereas mithramycin-a, an inhibitor of Sp1-like function, dose dependently blocked NGF and Sp1-dependent promoter activity in PC12 cells. Co-transfection of siRNA directed against Sp1 or Sp4 decreased promoter activity in DRG neurons and NGF treated PC12 cells. Finally, electroporation of Sp1 or Sp4 cDNA into cultures of DRG neurons directed an increase in Sp1/Sp4 mRNA and importantly an increase in TRPV1 mRNA. Conversely, combined si-RNA directed knockdown of Sp1/Sp4 resulted in a decrease in TRPV1 mRNA. CONCLUSION: Based on these studies, we now propose a model of TRPV1 expression that is dependent on Sp1-like transcription factors with Sp4 playing a predominant role in activating TRPV1 RNA transcription in DRG neurons. Given that increases of TRPV1 expression have been implicated in a wide range of pathophysiologic states including persistent painful conditions, blockade of Sp1 like transcription factors represents a novel direction in therapeutic strategies. PMID- 21645330 TI - Improving delirium care in the intensive care unit: the design of a pragmatic study. AB - BACKGROUND: Delirium prevalence in the intensive care unit (ICU) is high. Numerous psychotropic agents are used to manage delirium in the ICU with limited data regarding their efficacy or harms. METHODS/DESIGN: This is a randomized controlled trial of 428 patients aged 18 and older suffering from delirium and admitted to the ICU of Wishard Memorial Hospital in Indianapolis. Subjects assigned to the intervention group will receive a multicomponent pharmacological management protocol for delirium (PMD) and those assigned to the control group will receive no change in their usual ICU care. The primary outcomes of the trial are (1) delirium severity as measured by the Delirium Rating Scale revised-98 (DRS-R-98) and (2) delirium duration as determined by the Confusion Assessment Method for the ICU (CAM-ICU). The PMD protocol targets the three neurotransmitter systems thought to be compromised in delirious patients: dopamine, acetylcholine, and gamma-aminobutyric acid. The PMD protocol will target the reduction of anticholinergic medications and benzodiazepines, and introduce a low-dose of haloperidol at 0.5-1 mg for 7 days. The protocol will be delivered by a combination of computer (artificial intelligence) and pharmacist (human intelligence) decision support system to increase adherence to the PMD protocol. DISCUSSION: The proposed study will evaluate the content and the delivery process of a multicomponent pharmacological management program for delirium in the ICU. TRIAL REGISTRATION: ClinicalTrials.gov: NCT00842608. PMID- 21645332 TI - Metabolic manipulation in chronic heart failure: study protocol for a randomised controlled trial. AB - BACKGROUND: Heart failure is a major cause of morbidity and mortality in society. Current medical therapy centres on neurohormonal modulation with angiotensin converting enzyme inhibitors and beta-blockers. There is growing evidence for the use of metabolic manipulating agents as adjunctive therapy in patients with heart failure. We aim to determine the effect of perhexiline on cardiac energetics and alterations in substrate utilisation in patients with non-ischaemic dilated cardiomyopathy. METHODS: A multi-centre, prospective, randomised double-blind, placebo-controlled trial of 50 subjects with non-ischaemic dilated cardiomyopathy recruited from University Hospital Birmingham NHS Foundation Trust and Cardiff and Vale NHS Trust. Baseline investigations include magnetic resonance spectroscopy to assess cardiac energetic status, echocardiography to assess left ventricular function and assessment of symptomatic status. Subjects are then randomised to receive 200 mg perhexiline maleate or placebo daily for 4 weeks with serum drug level monitoring. All baseline investigations will be repeated at the end of the treatment period. A subgroup of patients will undergo invasive investigations with right and left heart catheterisation to calculate respiratory quotient, and mechanical efficiency. The primary endpoint is an improvement in the phosphocreatine to adenosine triphosphate ratio at 4 weeks. Secondary end points are: i) respiratory quotient; ii) mechanical efficiency; iii) change in left ventricular (LV) function. TRIAL REGISTRATION: ClinicalTrials.gov: NCT00841139 ISRCTN: ISRCTN72887836. PMID- 21645333 TI - Dysphoric milk ejection reflex: A case report. AB - Dysphoric Milk Ejection Reflex (D-MER) is an abrupt emotional "drop" that occurs in some women just before milk release and continues for not more than a few minutes. The brief negative feelings range in severity from wistfulness to self loathing, and appear to have a physiological cause. The authors suggest that an abrupt drop in dopamine may occur when milk release is triggered, resulting in a real or relative brief dopamine deficit for affected women. Clinicians can support women with D-MER in several ways; often, simply knowing that it is a recognized phenomenon makes the condition tolerable. Further study is needed. PMID- 21645334 TI - Interactions between human immunodeficiency virus (HIV)-1 Vpr expression and innate immunity influence neurovirulence. AB - BACKGROUND: Viral diversity and abundance are defining properties of human immunodeficiency virus (HIV)-1's biology and pathogenicity. Despite the increasing availability of antiretroviral therapy, HIV-associated dementia (HAD) continues to be a devastating consequence of HIV-1 infection of the brain although the underlying disease mechanisms remain uncertain. Herein, molecular diversity within the HIV-1 non-structural gene, Vpr, was examined in RNA sequences derived from brain and blood of HIV/AIDS patients with or without HIV associated dementia (HAD) together with the ensuing pathobiological effects. RESULTS: Cloned brain- and blood-derived full length vpr alleles revealed that amino acid residue 77 within the brain-derived alleles distinguished HAD (77Q) from non-demented (ND) HIV/AIDS patients (77R) (p < 0.05) although vpr transcripts were more frequently detected in HAD brains (p < 0.05). Full length HIV-1 clones encoding the 77R-ND residue induced higher IFN-alpha, MX1 and BST-2 transcript levels in human glia relative to the 77Q-HAD encoding virus (p < 0.05) but both viruses exhibited similar levels of gene expression and replication. Myeloid cells transfected with 77Q-(pVpr77Q-HAD), 77R (pVpr77R-ND) or Vpr null (pVpr(-))-containing vectors showed that the pVpr77R-ND vector induced higher levels of immune gene expression (p < 0.05) and increased neurotoxicity (p < 0.05). Vpr peptides (amino acids 70-96) containing the 77Q-HAD or 77R-ND motifs induced similar levels of cytosolic calcium activation when exposed to human neurons. Human glia exposed to the 77R-ND peptide activated higher transcript levels of IFN-alpha, MX1, PRKRA and BST-2 relative to 77Q-HAD peptide (p < 0.05). The Vpr 77R-ND peptide was also more neurotoxic in a concentration-dependent manner when exposed to human neurons (p < 0.05). Stereotaxic implantation of full length Vpr, 77Q-HAD or 77R-ND peptides into the basal ganglia of mice revealed that full length Vpr and the 77R-ND peptide caused greater neurobehavioral deficits and neuronal injury compared with 77Q-HAD peptide-implanted animals (p < 0.05). CONCLUSIONS: These observations underscored the potent neuropathogenic properties of Vpr but also indicated viral diversity modulates innate neuroimmunity and neurodegeneration. PMID- 21645335 TI - Primary care utilisation patterns among an urban immigrant population in the Spanish National Health System. AB - BACKGROUND: There is evidence suggesting that the use of health services is lower among immigrants after adjusting for age and sex. This study takes a step forward to compare primary care (PC) utilisation patterns between immigrants and the native population with regard to their morbidity burden. METHODS: This retrospective, observational study looked at 69,067 individuals representing the entire population assigned to three urban PC centres in the city of Zaragoza (Aragon, Spain). Poisson models were applied to determine the number of annual PC consultations per individual based on immigration status. All models were first adjusted for age and sex and then for age, sex and case mix (ACG System(r)). RESULTS: The age and sex adjusted mean number of total annual consultations was lower among the immigrant population (children: IRR = 0.79, p < 0.05; adults: IRR = 0.73, p < 0.05). After adjusting for morbidity burden, this difference decreased among children (IRR = 0.94, p < 0.05) and disappeared among adults (IRR = 1.00). Further analysis considering the PC health service and type of visit revealed higher usage of routine diagnostic tests among immigrant children (IRR = 1.77, p < 0.05) and a higher usage of emergency services among the immigrant adult population (IRR = 1.2, p < 0.05) after adjusting for age, sex and case mix. CONCLUSIONS: Although immigrants make lower use of PC services than the native population after adjusting the consultation rate for age and sex, these differences decrease significantly when considering their morbidity burden. These results reinforce the 'healthy migration effect' and discount the existence of differences in PC utilisation patterns between the immigrant and native populations in Spain. PMID- 21645336 TI - Gender bias revisited: new insights on the differential management of chest pain. AB - BACKGROUND: Chest pain is a common complaint and reason for consultation in primary care. Few data exist from a primary care setting whether male patients are treated differently than female patients. We examined whether there are gender differences in general physicians' (GPs) initial assessment and subsequent management of patients with chest pain, and how these differences can be explained METHODS: We conducted a prospective study with 1212 consecutive chest pain patients. The study was conducted in 74 primary care offices in Germany from October 2005 to July 2006. After a follow up period of 6 months, an independent interdisciplinary reference panel reviewed clinical data of every patient and decided about the etiology of chest pain at the time of patient recruitment (delayed type-reference standard). We adjusted gender differences of six process indicators for different models. RESULTS: GPs tended to assume that CHD is the cause of chest pain more often in male patients and referred more men for an exercise test (women 4.1%, men 7.3%, p = 0.02) and to the hospital (women 2.9%, men 6.6%, p < 0.01). These differences remained when adjusting for age and cardiac risk factors but ceased to exist after adjusting for the typicality of chest pain. CONCLUSIONS: While observed gender differences can not be explained by differences in age, CHD prevalence, and underlying risk factors, the less typical symptom presentation in women might be an underlying factor. However this does not seem to result in suboptimal management in women but rather in overuse of services for men. We consider our conclusions rather hypothesis generating and larger studies will be necessary to prove our proposed model. PMID- 21645337 TI - Pulmonary sclerosing hemangioma in a 21-year-old male with metastatic hereditary non-polyposis colorectal cancer: report of a case. AB - BACKGROUND: Pulmonary sclerosing hemangioma (SH) is a rare tumor of the lung predominantly affecting Asian women in their fifth decade of life. SH is thought to evolve from primitive respiratory epithelium and mostly shows benign biological behavior; however, cases of lymph node metastases, local recurrence and multiple lesions have been described. CASE PRESENTATION: We report the case of a 21-year-old Caucasian male with a history of locally advanced and metastatic rectal carcinoma (UICC IV; pT4, pN1, M1(hep)) that was eventually identified as having hereditary non-polyposis colorectal cancer (HNPCC, Lynch syndrome). After neoadjuvant chemotherapy followed by low anterior resection, adjuvant chemotherapy and metachronous partial hepatectomy, he was admitted for treatment of newly diagnosed bilateral pulmonary metastases. Thoracic computed tomography showed a homogenous, sharply marked nodule in the left lower lobe. We decided in favor of atypical resection followed by systematic lymphadenectomy. Histopathological analysis revealed the diagnosis of SH. CONCLUSIONS: Cases have been published with familial adenomatous polyposis (FAP) and simultaneous SH. FAP, Gardner syndrome and Li-Fraumeni syndrome, however, had been ruled out in the present case. To the best of our knowledge, this is the first report describing SH associated with Lynch syndrome. PMID- 21645338 TI - Rapid, progressive neuropathic arthropathy of the hip in a patient co-infected with human immunodeficiency virus, hepatitis C virus and tertiary syphilis: case report. AB - BACKGROUND: Syphilis is a chronic infection that is classified into three stages. In its tertiary stage, syphilis spreads to the brain, heart and other organs; the lesions may involve the skin, mucous membranes and bones. Neuropathic arthropathy associated with tertiary syphilis has rarely been described in Europe and its association with HIV-HCV co-infection has not been reported so far.This article reports the case of a man with tertiary syphilis presenting with rapidly evolving neuropathic arthropathy of the hip and extensive bone destruction. CASE PRESENTATION: On initial presentation, the patient complained of progressively worsening left-sided coxalgia without localized or generalized inflammation. The patient reported to have no history of previous infections, trauma or cancer. Plain x-ray films of the left coxofemoral joint showed marked degeneration with necrosis of the proximal epiphysis of femur and morphological alterations of the acetabulum without protrusion. Primary coxarthrosis was diagnosed and hip arthroplasty was offered, but the patient declined treatment. Three months later, the patient presented a marked deterioration of his general condition. He disclosed that he was seropositive for HCV and HIV, as confirmed by serology. Syphilis serology testing was also positive. A Girdlestone's procedure was performed and samples were collected for routine cultures for bacteria and acid fast bacilli, all resulting negative.Although histological findings were inconclusive, confirmed positive serology for syphilis associated with progressive arthropathy was strongly suggestive of tertiary syphilis, probably exacerbated by HIV-HCV co-infection. The patient partially recovered the ability to walk. CONCLUSIONS: Due to the resurgence of syphilis, this disease should be considered as a possible cause of neuropathic arthropathy when other infectious causes have been ruled out, particularly in patients with HIV and/or HCV co infection. PMID- 21645339 TI - Intra-familial and ethnic effects on attitudinal and perceptual body image: a cohort of South African mother-daughter dyads. AB - BACKGROUND: International studies suggest ethnic differences in obesity prevalence may be due, in part, to differences in body image and body size dissatisfaction between groups. Further, there is evidence to suggest that there is a familial resemblance in body image between mothers and their younger (preadolescent) daughters. This research was therefore conducted to specifically identify the extent to which family status (presented as mother-daughter resemblance) and ethnicity impact on body image attitudes and perceptions of South African mothers and their pre-adolescent daughters. METHODS: Mother daughter dyads (n = 201, 31% black, 37% mixed ancestry and 32% white) answered questions regarding their body image perception (the way they saw their body size status), their body image ideals, and body image attitudes (body size dissatisfaction in particular, presented as the Feel-Ideal Difference [FID] index score). Mothers' and daughters' body image results were compared within dyads and across ethnic groups using repeated measures of ANOVA. RESULTS: Overall, body image resemblances exist between South African mothers and their pre-adolescent daughters. Mothers and daughters chose similarly weighted silhouettes to represent their body size ideals (p = 0.308), regardless of their ethnicity or body mass index (BMI). The FID index scores were similar between mothers and their daughters only after the confounding effects of maternal BMI were removed (p = 0.685). The silhouettes chosen to represent thinness were also similar between mothers and their daughters (p = 0.960) regardless of ethnicity and maternal BMI. On the other hand, the silhouettes chosen to represent fatness were similar (p = 0.342) between mothers and their daughters, only after the confounding effects of maternal BMI were removed. Lastly, mothers and their daughters chose similarly weighted silhouettes as engendering feelings of beauty, respect and happiness (p = 0.813; p = 0.615 and p = 0.693, respectively). In this instance, black mother-daughter dyads chose significantly heavier silhouettes than the other ethnic groups. This implies that black mothers and daughters associate beauty, respect and happiness with a bigger body size. CONCLUSION: Resemblances exist between pre-adolescent girls and their mothers on issues related to ideal and attitudinal body image. In this regard, South African researchers should consider the effects ethnicity and family status on body image of women when developing targeted interventions to prevent or manage obesity. PMID- 21645340 TI - Angioplasty in acute middle cerebral artery stroke due to atrial fibrillation selected by CT perfusion: a case report. AB - We report the experience of a case of acute stroke in a patient affected by Rendu Osler syndrome and atrial fibrillation. The combination of dynamic computerized tomography perfusion scans and the use of a high-compliance balloon allowed increasing the treatment window for intra-arterial recanalization over 6 h after stroke onset in a patient with middle cerebral artery occlusion. PMID- 21645341 TI - siRNAs: potential therapeutic agents against hepatitis C virus. AB - Hepatitis C virus is a major cause of chronic liver diseases which can lead to permanent liver damage, hepatocellular carcinoma and death. The presently available treatment with interferon plus ribavirin, has limited benefits due to adverse side effects such as anemia, depression and "flu-like" symptoms. Needless to mention, the effectiveness of interferon therapy is predominantly, if not exclusively, limited to virus type 3a and 3b whereas in Europe and North America the majority of viral type is 1a and 2a. Due to the limited efficiency of current therapy, RNA interference (RNAi) a novel regulatory and powerful silencing approach for molecular therapeutics through a sequence-specific RNA degradation process represents an alternative option. Several reports have indicated the efficiency and specificity of synthetic and vector based siRNAs inhibiting HCV replication. In the present review, we focused that combination of siRNAs against virus and host genes will be a better option to treat HCV. PMID- 21645342 TI - Toxic marine microalgae and shellfish poisoning in the British isles: history, review of epidemiology, and future implications. AB - The relationship between toxic marine microalgae species and climate change has become a high profile and well discussed topic in recent years, with research focusing on the possible future impacts of changing hydrological conditions on Harmful Algal Bloom (HAB) species around the world. However, there is very little literature concerning the epidemiology of these species on marine organisms and human health. Here, we examine the current state of toxic microalgae species around the UK, in two ways: first we describe the key toxic syndromes and gather together the disparate reported data on their epidemiology from UK records and monitoring procedures. Secondly, using NHS hospital admissions and GP records from Wales, we attempt to quantify the incidence of shellfish poisoning from an independent source. We show that within the UK, outbreaks of shellfish poisoning are rare but occurring on a yearly basis in different regions and affecting a diverse range of molluscan shellfish and other marine organisms. We also show that the abundance of a species does not necessarily correlate to the rate of toxic events. Based on routine hospital records, the numbers of shellfish poisonings in the UK are very low, but the identification of the toxin involved, or even a confirmation of a poisoning event is extremely difficult to diagnose. An effective shellfish monitoring system, which shuts down aquaculture sites when toxins exceed regularity limits, has clearly prevented serious impact to human health, and remains the only viable means of monitoring the potential threat to human health. However, the closure of these sites has an adverse economic impact, and the monitoring system does not include all toxic plankton. The possible geographic spreading of toxic microalgae species is therefore a concern, as warmer waters in the Atlantic could suit several species with southern biogeographical affinities enabling them to occupy the coastal regions of the UK, but which are not yet monitored or considered to be detrimental. PMID- 21645343 TI - Weight Watchers on prescription: an observational study of weight change among adults referred to Weight Watchers by the NHS. AB - BACKGROUND: The scale of overweight and obesity in the UK places a considerable burden on the NHS. In some areas the NHS has formed partnerships with commercial companies to offer weight management services, but there has been little evaluation of these schemes.This study is an independent audit of the Weight Watchers NHS Referral scheme and evaluates the weight change of obese and overweight adults referred to Weight Watchers (WW) by the NHS. METHOD: Data was obtained from the WW NHS Referral Scheme database for 29,326 referral courses started after 2nd April 2007 and ending before 6th October 2009 [90% female; median age 49 years (IQR 38-61 years); median BMI 35.1 kg/m2 (IQR 31.8-39.5 kg/m2). Participants received vouchers (funded by the PCT following referral by a healthcare professional) to attend 12 WW meetings. Body weight was measured at WW meetings and relayed to the central database. RESULTS: Median weight change for all referrals was -2.8 kg [IQR -5.9--0.7 kg] representing -3.1% initial weight. 33% of all courses resulted in loss of >=5% initial weight. 54% of courses were completed. Median weight change for those completing a first course was -5.4 kg [IQR -7.8--3.1 kg] or -5.6% of initial weight. 57% lost >=5% initial weight. CONCLUSIONS: A third of all patients who were referred to WW through the WW NHS Referral Scheme and started a 12 session course achieved >=5% weight loss, which is usually associated with clinical benefits. This is the largest audit of NHS referral to a commercial weight loss programme in the UK and results are comparable with other options for weight loss available through primary care. PMID- 21645344 TI - The role of lifestyle changes in the management of chronic liver disease. AB - The prevalence of obesity worldwide has dramatically increased during the last three decades. With obesity comes a variety of adverse health outcomes which are grouped under the umbrella of metabolic syndrome. The liver in particular seems to be significantly impacted by fat deposition in the presence of obesity. In this article we discuss several liver conditions which are directly affected by overweight and obese status, including non-alcoholic fatty liver disease, chronic infection with hepatitis C virus and post-liver transplant status. The deleterious effects of obesity on liver disease and overall health can be significantly impacted by a culture that fosters sustained nutritional improvement and regular physical activity. Here we summarize the current evidence supporting non-pharmacological, lifestyle interventions that lead to weight reduction, improved physical activity and better nutrition as part of the management and treatment of these liver conditions. PMID- 21645345 TI - Scaling properties of protein family phylogenies. AB - BACKGROUND: One of the classical questions in evolutionary biology is how evolutionary processes are coupled at the gene and species level. With this motivation, we compare the topological properties (mainly the depth scaling, as a characterization of balance) of a large set of protein phylogenies with those of a set of species phylogenies. RESULTS: The comparative analysis between protein and species phylogenies shows that both sets of phylogenies share a remarkably similar scaling behavior, suggesting the universality of branching rules and of the evolutionary processes that drive biological diversification from gene to species level. In order to explain such generality, we propose a simple model which allows us to estimate the proportion of evolvability/robustness needed to approximate the scaling behavior observed in the phylogenies, highlighting the relevance of the robustness of a biological system (species or protein) in the scaling properties of the phylogenetic trees. CONCLUSIONS: The invariance of the scaling properties at levels spanning from genes to species suggests that rules that govern the incapability of a biological system to diversify are equally relevant both at the gene and at the species level. PMID- 21645346 TI - Paula Modersohn-Becker, the challenges of pregnancy and the weight of tradition. AB - Paula Modersohn-Becker, widely considered to have been one of the most important independent Expressionist painters of the early twentieth century, was thirty-one years old when she gave birth to her first child. Following the then-common practice of putting women to bed rest for two-four weeks after delivery, she died of massive pulmonary embolism when she was first allowed to stand, eighteen days after giving birth. Paula had foreseen her death at a young age and was apprehensive about her pregnancy, yet she painted herself as pregnant in her best known self-portrait, thus underlining the importance of the pregnancy in her life. In the light of knowledge available at the time, the authors present a brief discussion of the life and death of Paula Modersohn-Becker as a reflection on the potential dangers of blindly following conventional wisdom in the medical profession. PMID- 21645347 TI - Opportunistic screening for skin cancer using a mobile unit in Brazil. AB - BACKGROUND: Skin cancer is the most common malignancy in the white population worldwide. In Brazil, the National Cancer Institute (INCA) estimates that in 2010 there will be 119,780 and 5,930 new cases of non-melanoma skin cancer and melanoma, respectively. The aim of this study was to evaluate the use of a mobile unit in the diagnosis and treatment of skin cancer in several poor regions of Brazil. METHODS: The diagnosis of skin cancer was accomplished through active medical screening in the prevention Mobile Unit (MU) of Barretos Cancer Hospital (BCH). The study population consisted of patients examined in the MU between 2004 and 2007, and their suspicious lesions were subjected to histopathological evaluation. Data were collected prospectively from standardized forms and analyzed. RESULTS: During the screening, 17,857 consultations were carried out. A total of 2012 (11.2%) cases of skin cancer were diagnosed. The predominant histological type reported was basal cell carcinoma (n = 1,642 or 81.6%), followed by squamous cell carcinoma (n = 303 or 15.1%), Bowen's disease (n = 25 or 1.2%), malignant melanoma (n = 23 or 1.1%), basosquamous cell carcinoma (n = 3 or 0.1%), miscellaneous lesions (12 or 0.6%), and metatypical carcinoma (n = 4 or 0.2%). Only 0.6% of lesions were stage III. There were no stage IV non-melanoma skin lesions, as well as no melanomas stages III and IV, found. CONCLUSIONS: It was observed that the MU can be a useful tool for early skin cancer diagnosis and treatment. This program probably is important, especially in developing countries with inadequate public health systems and social inequality. PMID- 21645348 TI - Comparison of 1.5T and 3T MRI scanners in evaluation of acute bone stress in the foot. AB - BACKGROUND: Bone stress injuries are common in athletes and military recruits. Only a minority of bone stress changes are available on plain radiographs. Acute bone stress is often visible on MRI as bone marrow edema, which is also seen in many other disease processes such as malignancies, inflammatory conditions and infections. The purpose of this study was to investigate the ability of radiographs, 1.5T and 3T MRI to identify acute bone marrow changes in the foot. METHODS: Ten patients with 12 stress fractures seen on plain radiographs underwent MRI using 1.5T and 3T scanners. T1 FSE and STIR axial, sagittal, and coronal view sequences were obtained. Two musculoskeletal radiologists interpreted the images independently and by consensus in case of disagreement. RESULTS: Of the 63 acute bone stress changes seen on 3T images, 61 were also seen on 1.5T images. The sensitivity of 1.5T MRI was 97% (95% CI: 89%-99%) compared with 3T. The 3T MRI images where, therefore, at least equally sensitive to 1.5T scanners in detection of bone marrow edema. On T1-weighted sequences, 3T images were slightly superior to 1.5T images in visualizing the demarcation of the edema and bone trabeculae. The kappa-value for inter-observer variability was 0.86 in the MRI indicating substantial interobserver agreement. CONCLUSIONS: Owing to slightly better resolution of 3T images, edema characterization is easier, which might aid in the differential diagnosis of the bone marrow edema. There was, however, no noteworthy difference in the sensitivity of the 1.5T and 3T images to bone marrow edema. Routine identification of acute bone stress changes and suspected stress injuries can, therefore, be made with 1.5T field strength. PMID- 21645349 TI - Transport processes of radiopharmaceuticals and -modulators. AB - Radiotherapy and radiology have been indispensable components in cancer care for many years. The detection limit of small tumor foci as well as the development of radio-resistance and severe side effects towards normal tissues led to the development of strategies to improve radio-diagnostic and -therapeutic approaches by pharmaceuticals. The term "radiopharmaceutical" has been used for drugs labeled with radioactive tracers for therapy or diagnosis. In addition, drugs have been described to sensitize tumor cells to radiotherapy (radiosensitizers) or to protect normal tissues from detrimental effects of radiation (radioprotectors). The present review summarizes recent concepts on the transport of radiopharmaceuticals, radiosensitizers, and radioprotectors in cells and tissues, e.g. by ATP-binding cassette transporters such as P-glycoprotein. Strengths and weaknesses of current strategies to improve transport-based processes are discussed. PMID- 21645350 TI - Impact of timing of renal replacement therapy initiation on outcome of septic acute kidney injury. AB - INTRODUCTION: Sepsis is the leading cause of acute kidney injury (AKI) in critical patients. The optimal timing of initiating renal replacement therapy (RRT) in septic AKI patients remains controversial. The objective of this study is to determine the impact of early or late initiation of RRT, as defined using the simplified RIFLE (risk, injury, failure, loss of kidney function, and end stage renal failure) classification (sRIFLE), on hospital mortality among septic AKI patients. METHODS: Patient with sepsis and AKI requiring RRT in surgical intensive care units were enrolled between January 2002 and October 2009. The patients were divided into early (sRIFLE-0 or -Risk) or late (sRIFLE-Injury or Failure) initiation of RRT by sRIFLE criteria. Cox proportional hazard ratios for in hospital mortality were determined to assess the impact of timing of RRT. RESULTS: Among the 370 patients, 192 (51.9%) underwent early RRT and 259 (70.0%) died during hospitalization. The mortality rate in early and late RRT groups were 70.8% and 69.7% respectively (P > 0.05). Early dialysis did not relate to hospital mortality by Cox proportional hazard model (P > 0.05). Patients with heart failure, male gender, higher admission creatinine, and operation were more likely to be in the late RRT group. Cox proportional hazard model, after adjustment with propensity score including all patients based on the probability of late RRT, showed early dialysis was not related to hospital mortality. Further model matched patients by 1:1 fashion according to each patient's propensity to late RRT showed no differences in hospital mortality according to head-to-head comparison of demographic data (P > 0.05). CONCLUSIONS: Use of sRIFLE classification as a marker poorly predicted the benefits of early or late RRT in the context of septic AKI. In the future, more physiologically meaningful markers with which to determine the optimal timing of RRT initiation should be identified. PMID- 21645351 TI - Synergistic effects of oncolytic reovirus and docetaxel chemotherapy in prostate cancer. AB - BACKGROUND: Reovirus type 3 Dearing (T3D) has demonstrated oncolytic activity in vitro, in in vivo murine models and in early clinical trials. However the true potential of oncolytic viruses may only be realized fully in combination with other modalities such as chemotherapy, targeted therapy and radiotherapy. In this study, we examine the oncolytic activity of reovirus T3D and chemotherapeutic agents against human prostate cancer cell lines, with particular focus on the highly metastatic cell line PC3 and the chemotherapeutic agent docetaxel. Docetaxel is the standard of care for metastatic prostate cancer and acts by disrupting the normal process of microtubule assembly and disassembly. Reoviruses have been shown to associate with microtubules and may require this association for efficient viral replication. METHODS: The effects of reovirus and chemotherapy on in vitro cytotoxicity were investigated in PC3 and Du 145 cells and the interactions between agents were assessed by combination index analysis. An Annexin V/propidium iodide fluorescence-activated cell sorting-based assay was used to determine mode of cell death. The effects of reovirus and docetaxel administered as single agent or combination therapy were tested in vivo in a murine model. The effects of docetaxel and reovirus, alone and together, on microtubule stabilisation were investigated by Western blot analysis. RESULTS: Variable degrees of synergistic cytotoxicity were observed in PC3 and Du 145 cells exposed to live reovirus and several chemotherapy agents. Combination of reovirus infection with docetaxel exposure led to increased late apoptotic/necrotic cell populations. Reovirus/docetaxel combined therapy led to reduced tumour growth and increased survival in a PC3 tumour bearing mouse model. Microtubule stabilization was enhanced in PC3 cells treated with reovirus/docetaxel combined therapy compared to other reovirus/chemotherapy combinations. CONCLUSIONS: The co-administration of a variety of chemotherapeutic agents with live reovirus was able to enhance cytotoxicity synergistically in vitro. The combination of docetaxel with reovirus also delayed tumour growth and improved survival in vivo. Enhanced microtubule stabilisation following this combination treatment may, in part, explain the mechanism of synergy. These results provide evidence to support the ongoing clinical trials using these agents. PMID- 21645352 TI - Severe acquired cytomegalovirus infection in a full-term, formula-fed infant: case report. AB - BACKGROUND: Cases of cytomegalovirus colitis are exceptionally reported in immuno competent infant. The pathogenesis is uncertain but breast-feeding is considered as a main source of postnatal infection. CASE PRESENTATION: Here we report a full term, formula-fed infant who developed a severe cytomegalovirus anaemia and colitis when aged 2 months. CONCLUSION: Even if the molecular identity between the cytomegalovirus-isolate of the infant and the maternal virus could not be demonstrated, we confirmed through laboratory investigation that cytomegalovirus infection was acquired postnatally. However, the source of cytomegalovirus infection remained unclear. Alternative modes of cytomegalovirus transmission are discussed. PMID- 21645353 TI - Optimum conductive fabric sensor sites for evaluating the status of knee joint movements using bio-impedance. AB - BACKGROUND: There have been many studies that utilize the bio-impedance measurement method to analyze the movements of the upper and lower limbs. A fixed electrical current flows into the limbs through four standard disposable electrodes in this method. The current flows in the muscles and blood vessels, which have relatively low resistivity levels in the human body. This method is used to measure bio-impedance changes following volume changes of muscles and blood vessels around a knee joint. The result of the bio-impedance changes is used to evaluate the movements. However, the method using the standard disposable electrodes has a restriction related to its low bio-impedance changes: the standard disposable electrodes are only able to measure bio-impedance from a limited part of a muscle. Moreover, it is impossible to use continuously, as the electrodes are designed to be disposable. This paper describes a conductive fabric sensor (CFS) using a bio-impedance measurement method and determines the optimum configuration of the sensor for estimating knee joint movements. METHODS: The upper side of subjects' lower limbs was divided into two areas and the lower side of subjects' lower limbs was divided into three areas. The spots were matched and 6 pairs were selected. Subjects were composed of 15 males (age: 30.7 +/- 5.3, weight: 69.8 +/- 4.2 kg, and height: 173.5 +/- 2.8 cm) with no known problems with their knee joints. Bio-impedance changes according to knee joint flexion/extension assessments were calculated and compared with bio-impedance changes by an ankle joint flexion/extension test (SNR I) and a hip joint flexion/extension test (SNR II). RESULTS: The bio-impedance changes of the knee joint flexion/extension assessment were 35.4 +/- 20.0 Omega on the (1, 5) pair. SNR I was 3.8 +/- 8.4 and SNR II was 6.6 +/- 7.9 on the (1, 5) pair. CONCLUSIONS: The optimum conductive fabric sensor configuration for evaluating knee joint movements were represented by the (1, 5) pair. PMID- 21645354 TI - Automated data extraction from general practice records in an Australian setting: trends in influenza-like illness in sentinel general practices and emergency departments. AB - BACKGROUND: Influenza intelligence in New South Wales (NSW), Australia is derived mainly from emergency department (ED) presentations and hospital and intensive care admissions, which represent only a portion of influenza-like illness (ILI) in the population. A substantial amount of the remaining data lies hidden in general practice (GP) records. Previous attempts in Australia to gather ILI data from GPs have given them extra work. We explored the possibility of applying automated data extraction from GP records in sentinel surveillance in an Australian setting.The two research questions asked in designing the study were: Can syndromic ILI data be extracted automatically from routine GP data? How do ILI trends in sentinel general practice compare with ILI trends in EDs? METHODS: We adapted a software program already capable of automated data extraction to identify records of patients with ILI in routine electronic GP records in two of the most commonly used commercial programs. This tool was applied in sentinel sites to gather retrospective data for May-October 2007-2009 and in real-time for the same interval in 2010. The data were compared with that provided by the Public Health Real-time Emergency Department Surveillance System (PHREDSS) and with ED data for the same periods. RESULTS: The GP surveillance tool identified seasonal trends in ILI both retrospectively and in near real-time. The curve of seasonal ILI was more responsive and less volatile than that of PHREDSS on a local area level. The number of weekly ILI presentations ranged from 8 to 128 at GP sites and from 0 to 18 in EDs in non-pandemic years. CONCLUSION: Automated data extraction from routine GP records offers a means to gather data without introducing any additional work for the practitioner. Adding this method to current surveillance programs will enhance their ability to monitor ILI and to detect early warning signals of new ILI events. PMID- 21645355 TI - Measurement properties of translated versions of neck-specific questionnaires: a systematic review. AB - BACKGROUND: Several disease-specific questionnaires to measure pain and disability in patients with neck pain have been translated. However, a simple translation of the original version doesn't guarantee similar measurement properties. The objective of this study is to critically appraise the quality of the translation process, cross-cultural validation and the measurement properties of translated versions of neck-specific questionnaires. METHODS: Bibliographic databases were searched for articles concerning the translation or evaluation of the measurement properties of a translated version of a neck-specific questionnaire. The methodological quality of the selected studies and the results of the measurement properties were critically appraised and rated using the COSMIN checklist and criteria for measurement properties. RESULTS: The search strategy resulted in a total of 3641 unique hits, of which 27 articles, evaluating 6 different questionnaires in 15 different languages, were included in this study. Generally the methodological quality of the translation process is poor and none of the included studies performed a cross-cultural adaptation. A substantial amount of information regarding the measurement properties of translated versions of the different neck-specific questionnaires is lacking. Moreover, the evidence for the quality of measurement properties of the translated versions is mostly limited or assessed in studies of poor methodological quality. CONCLUSIONS: Until results from high quality studies are available, we advise to use the Catalan, Dutch, English, Iranian, Korean, Spanish and Turkish version of the NDI, the Chinese version of the NPQ, and the Finnish, German and Italian version of the NPDS. The Greek NDI needs cross-cultural validation and there is no methodologically sound information for the Swedish NDI. For all other languages we advise to translate the original version of the NDI. PMID- 21645356 TI - The interplay between surfaces and soluble factors define the immunologic and angiogenic properties of myeloid dendritic cells. AB - BACKGROUND: Dendritic cells (DCs) are antigen presenting cells capable of inducing specific immune responses against microbial infections, transplant antigens, or tumors. Interestingly, microenvironment conditions such as those present in tumor settings might induce a DC phenotype that is poorly immunogenic and with the capability of promoting angiogenesis. We hypothesize that this plasticity may be caused not only by the action of specific cytokines or growth factors but also by the properties of the surfaces with which they interact, such as extracellular matrix (ECM) components. RESULTS: Herewith we studied the effect of different surfaces and soluble factors on the biology of DCs. To accomplish this, we cultured murine myeloid(m) DCs on surfaces coated with fibronectin, collagen I, gelatin, and Matrigel using poly-D-lysine and polystyrene as non biological surfaces. Further, we cultured these cells in the presence of regular DC medium (RPMI 10% FBS) or commercially available endothelial medium (EGM-2). We determined that mDCs could be kept in culture up to 3 weeks in these conditions, but only in the presence of GM-CSF. We were able to determine that long-term DC cultures produce an array of angiogenic factors, and that some of these cultures still retain the capability to induce T cell responses. CONCLUSIONS: Altogether these data indicate that in order to design DC-based vaccines or treatments focused on changing the phenotype of DCs associated with diseases such as cancer or atherosclerosis, it becomes necessary to fully investigate the microenvironment in which these cells are present or will be delivered. PMID- 21645357 TI - Analysis of BAC end sequences in oak, a keystone forest tree species, providing insight into the composition of its genome. AB - BACKGROUND: One of the key goals of oak genomics research is to identify genes of adaptive significance. This information may help to improve the conservation of adaptive genetic variation and the management of forests to increase their health and productivity. Deep-coverage large-insert genomic libraries are a crucial tool for attaining this objective. We report herein the construction of a BAC library for Quercus robur, its characterization and an analysis of BAC end sequences. RESULTS: The EcoRI library generated consisted of 92,160 clones, 7% of which had no insert. Levels of chloroplast and mitochondrial contamination were below 3% and 1%, respectively. Mean clone insert size was estimated at 135 kb. The library represents 12 haploid genome equivalents and, the likelihood of finding a particular oak sequence of interest is greater than 99%. Genome coverage was confirmed by PCR screening of the library with 60 unique genetic loci sampled from the genetic linkage map. In total, about 20,000 high-quality BAC end sequences (BESs) were generated by sequencing 15,000 clones. Roughly 5.88% of the combined BAC end sequence length corresponded to known retroelements while ab initio repeat detection methods identified 41 additional repeats. Collectively, characterized and novel repeats account for roughly 8.94% of the genome. Further analysis of the BESs revealed 1,823 putative genes suggesting at least 29,340 genes in the oak genome. BESs were aligned with the genome sequences of Arabidopsis thaliana, Vitis vinifera and Populus trichocarpa. One putative collinear microsyntenic region encoding an alcohol acyl transferase protein was observed between oak and chromosome 2 of V. vinifera. CONCLUSIONS: This BAC library provides a new resource for genomic studies, including SSR marker development, physical mapping, comparative genomics and genome sequencing. BES analysis provided insight into the structure of the oak genome. These sequences will be used in the assembly of a future genome sequence for oak. PMID- 21645358 TI - A step forward for understanding the morbidity burden in Guinea: a national descriptive study. AB - BACKGROUND: Little evidence on the burden of disease has been reported about Guinea. This study was conducted to demonstrate the morbidity burden in Guinea and provide basic evidence for setting health priorities. METHODS: A retrospective descriptive study was designed to present the morbidity burden of Guinea. Morbidity data were extracted from the National Health Statistics Report of Guinea of 2008. The data are collected based on a pyramid of facilities which includes two national hospitals (teaching hospitals), seven regional hospitals, 26 prefectural hospitals, 8 communal medical centers, 390 health centers, and 628 health posts. Morbidity rates were calculated to measure the burden of non-fatal diseases. The contributions of the 10 leading diseases were presented by sex and age group. RESULTS: In 2008, a total of 3,936,599 cases occurred. The morbidity rate for males was higher than for females, 461 versus 332 per 1,000 population. Malaria, respiratory infections, diarrheal diseases, helminthiases, and malnutrition ranked in the first 5 places and accounted for 74% of the total burden, respectively having a rate of 148, 64, 33, 32, and 14 per 1,000 population. The elderly aged 65+ had the highest morbidity rate (611 per 1,000 population) followed by working-age population (458 per 1,000 population) and children (396 per 1,000 population) while the working-age population aged 25-64 contributed the largest part (39%) to total cases. The sex- and age-specific spectrum of morbidity burden showed a similar profile except for small variations. CONCLUSION: Guinea has its unique morbidity burden. The ten leading causes of morbidity burden, especially for malaria, respiratory infections, diarrheal diseases, helminthiases, and malnutrition, need to be prioritized in Guinea. PMID- 21645359 TI - RNA-seq: technical variability and sampling. AB - BACKGROUND: RNA-seq is revolutionizing the way we study transcriptomes. mRNA can be surveyed without prior knowledge of gene transcripts. Alternative splicing of transcript isoforms and the identification of previously unknown exons are being reported. Initial reports of differences in exon usage, and splicing between samples as well as quantitative differences among samples are beginning to surface. Biological variation has been reported to be larger than technical variation. In addition, technical variation has been reported to be in line with expectations due to random sampling. However, strategies for dealing with technical variation will differ depending on the magnitude. The size of technical variance, and the role of sampling are examined in this manuscript. RESULTS: In this study three independent Solexa/Illumina experiments containing technical replicates are analyzed. When coverage is low, large disagreements between technical replicates are apparent. Exon detection between technical replicates is highly variable when the coverage is less than 5 reads per nucleotide and estimates of gene expression are more likely to disagree when coverage is low. Although large disagreements in the estimates of expression are observed at all levels of coverage. CONCLUSIONS: Technical variability is too high to ignore. Technical variability results in inconsistent detection of exons at low levels of coverage. Further, the estimate of the relative abundance of a transcript can substantially disagree, even when coverage levels are high. This may be due to the low sampling fraction and if so, it will persist as an issue needing to be addressed in experimental design even as the next wave of technology produces larger numbers of reads. We provide practical recommendations for dealing with the technical variability, without dramatic cost increases. PMID- 21645360 TI - Cell functional enviromics: unravelling the function of environmental factors. AB - BACKGROUND: While functional genomics, focused on gene functions and gene-gene interactions, has become a very active field of research in molecular biology, equivalent methodologies embracing the environment and gene-environment interactions are relatively less developed. Understanding the function of environmental factors is, however, of paramount importance given the complex, interactive nature of environmental and genetic factors across multiple time scales. RESULTS: Here, we propose a systems biology framework, where the function of environmental factors is set at its core. We set forth a "reverse" functional analysis approach, whereby cellular functions are reconstructed from the analysis of dynamic envirome data. Our results show these data sets can be mapped to less than 20 core cellular functions in a typical mammalian cell culture, while explaining over 90% of flux data variance. A functional enviromics map can be created, which provides a template for manipulating the environmental factors to induce a desired phenotypic trait. CONCLUSION: Our results support the feasibility of cellular function reconstruction guided by the analysis and manipulation of dynamic envirome data. PMID- 21645361 TI - Prevalence and risk factors of childhood allergic diseases in eight metropolitan cities in China: a multicenter study. AB - BACKGROUND: Several studies conducted during the past two decades suggested increasing trend of childhood allergic diseases in China. However, few studies have provided detailed description of geographic variation and explored risk factors of these diseases. This study investigated the pattern and risk factors of asthma, allergic rhinitis and eczema in eight metropolitan cities in China. METHODS: We conducted a cross-sectional survey during November-December 2005 in eight metropolitan cities in China. A total of 23791 children aged 6-13 years participated in this survey. Questions from the standard questionnaire of the International Study of Asthma and Allergies in Children (ISAAC) were used to examine the pattern of current asthma, allergic rhinitis and eczema. Logistic regression analyses were performed to assess the risk factors for childhood allergies. RESULTS: The average prevalence of childhood asthma, allergic rhinitis and eczema across the eight cities was 3.3% (95% Confidence interval (CI): 3.1%, 3.6%), 9.8% (95% CI: 9.4%, 10.2%) and 5.5% (95% CI: 5.2%, 5.8%), respectively. Factors related to lifestyle, mental health and socio-economic status were found to be associated with the prevalence of childhood allergies. These risk factors were unevenly distributed across cities and disproportionately affected the local prevalence. CONCLUSIONS: There was apparent geographic variation of childhood allergies in China. Socio-environmental factors had strong impacts on the prevalence of childhood allergies; but these impacts differed across regions. Thus public health policies should specifically target at the local risk factors for each individual area. PMID- 21645362 TI - Regulation of asymmetric cell division in the epidermis. AB - For proper tissue morphogenesis, cell divisions and cell fate decisions must be tightly and coordinately regulated. One elegant way to accomplish this is to couple them with asymmetric cell divisions. Progenitor cells in the developing epidermis undergo both symmetric and asymmetric cell divisions to balance surface area growth with the generation of differentiated cell layers. Here we review the molecular machinery implicated in controlling asymmetric cell division. In addition, we discuss the ability of epidermal progenitors to choose between symmetric and asymmetric divisions and the key regulatory points that control this decision. PMID- 21645363 TI - Generation of bivalent chromatin domains during cell fate decisions. AB - BACKGROUND: In self-renewing, pluripotent cells, bivalent chromatin modification is thought to silence (H3K27me3) lineage control genes while 'poising' (H3K4me3) them for subsequent activation during differentiation, implying an important role for epigenetic modification in directing cell fate decisions. However, rather than representing an equivalently balanced epigenetic mark, the patterns and levels of histone modifications at bivalent genes can vary widely and the criteria for identifying this chromatin signature are poorly defined. RESULTS: Here, we initially show how chromatin status alters during lineage commitment and differentiation at a single well characterised bivalent locus. In addition we have determined how chromatin modifications at this locus change with gene expression in both ensemble and single cell analyses. We also show, on a global scale, how mRNA expression may be reflected in the ratio of H3K4me3/H3K27me3. CONCLUSIONS: While truly 'poised' bivalently modified genes may exist, the original hypothesis that all bivalent genes are epigenetically premarked for subsequent expression might be oversimplistic. In fact, from the data presented in the present work, it is equally possible that many genes that appear to be bivalent in pluripotent and multipotent cells may simply be stochastically expressed at low levels in the process of multilineage priming. Although both situations could be considered to be forms of 'poising', the underlying mechanisms and the associated implications are clearly different. PMID- 21645364 TI - Loss of function mutations in the progranulin gene are related to pro inflammatory cytokine dysregulation in frontotemporal lobar degeneration patients. AB - The progranulin gene (PGRN) encodes a pleiotropic molecule with anti-inflammatory actions and neuronal protective effects. Accordingly, PGRN-deficient mice have been demonstrated to develop enhanced inflammation and progressive neurodegeneration. Loss of function mutations of the PGRN gene have been also reported to cause frontotemporal lobar degeneration (FTLD), a neurodegenerative disease leading to dementia generally in the presenium. Since neurodegeneration might be negatively impacted by chronic inflammation, the possible influence of PGRN defects on inflammatory pathways appears to be of great relevance for the understanding of neurodegeneration pathogenic processes in those patients. However, no data about the inflammatory profile of PGRN-defective subjects have been so far provided.In this study, we analyzed serum levels of the pro inflammatory mediators IL-6, TNF-alpha and IL-18 in FTLD patients with or without PGRN mutations, at both pre-symptomatic and symptomatic stages. We provide evidence that circulating IL-6 is increased in PGRN-mutated FTLD patients, as compared to both PGRN non-mutated FTLD patients and controls. In contrast, levels of IL-6 were not altered in asymptomatic subjects carrying the PGRN mutations. Finally, TNF-alpha and IL-18 serum levels did not differ among all groups of included subjects.We conclude that the profile of circulating pro-inflammatory cytokines is altered in PGRN-related symptomatic FTLD. Thus, our findings point to IL-6 as a possible specific mediator and a potential therapeutic target in this monogenic disease, suggesting that an enhanced inflammatory response might be indeed involved in its progression. PMID- 21645365 TI - Determination of the relative economic impact of different molecular-based laboratory algorithms for respiratory viral pathogen detection, including Pandemic (H1N1), using a secure web based platform. AB - BACKGROUND: During period of crisis, laboratory planners may be faced with a need to make operational and clinical decisions in the face of limited information. To avoid this dilemma, our laboratory utilizes a secure web based platform, Data Integration for Alberta Laboratories (DIAL) to make near real-time decisions.This manuscript utilizes the data collected by DIAL as well as laboratory test cost modeling to identify the relative economic impact of four proposed scenarios of testing for Pandemic H1N1 (2009) and other respiratory viral pathogens. METHODS: Historical data was collected from the two waves of the pandemic using DIAL. Four proposed molecular testing scenarios were generated: A) Luminex respiratory virus panel (RVP) first with/without US centers for Disease Control Influenza A Matrix gene assay (CDC-M), B) CDC-M first with/without RVP, C) RVP only, and D) CDC-M only. Relative cost estimates of different testing algorithm were generated from a review of historical costs in the lab and were based on 2009 Canadian dollars. RESULTS: Scenarios A and B had similar costs when the rate of influenza A was low (< 10%) with higher relative cost in Scenario A with increasing incidence. Scenario A provided more information about mixed respiratory virus infection as compared with Scenario B. CONCLUSIONS: No one approach is applicable to all conditions. Testing costs will vary depending on the test volume, prevalence of influenza A strains, as well as other circulating viruses and a more costly algorithm involving a combination of different tests may be chosen to ensure that tests results are returned to the clinician in a quicker manner. Costing should not be the only consideration for determination of laboratory algorithms. PMID- 21645366 TI - BMP signaling balances proliferation and differentiation of muscle satellite cell descendants. AB - BACKGROUND: The capacity of muscle to grow or to regenerate after damage is provided by adult stem cells, so called satellite cells, which are located under the basement lamina of each myofiber. Upon activation satellite cells enter the cell cycle, proliferate and differentiate into myoblasts, which fuse to injured myofibers or form new fibers. These processes are tightly controlled by many growth factors. RESULTS: Here we investigate the role of bone morphogenetic proteins (BMPs) during satellite cell differentiation. Unlike the myogenic C2C12 cell line, primary satellite cells do not differentiate into osteoblasts upon BMP signaling. Instead BMP signaling inhibits myogenic differentiation of primary satellite cells ex vivo. In contrast, inhibition of BMP signaling results in cell cycle exit, followed by enhanced myoblast differentiation and myotube formation. Using an in vivo trauma model we demonstrate that satellite cells respond to BMP signals during the regeneration process. Interestingly, we found the BMP inhibitor Chordin upregulated in primary satellite cell cultures and in regenerating muscles. In both systems Chordin expression follows that of Myogenin, a marker for cells committed to differentiation. CONCLUSION: Our data indicate that BMP signaling plays a critical role in balancing proliferation and differentiation of activated satellite cells and their descendants. Initially, BMP signals maintain satellite cells descendants in a proliferating state thereby expanding cell numbers. After cells are committed to differentiate they upregulate the expression of the BMP inhibitor Chordin thereby supporting terminal differentiation and myotube formation in a negative feedback mechanism. PMID- 21645367 TI - CyberKnife stereotactic radiosurgery for recurrent, metastatic, and residual hemangiopericytomas. AB - OBJECTIVE: Hemangiopericytoma is a rare and aggressive meningeal tumor. Although surgical resection is the standard treatment, hemangiopericytomas often recur with high incidences of metastasis. The purpose of this study was to evaluate the role of CyberKnife stereotactic radiosurgery (CK) in the management of recurrent, metastatic, and residual hemangiopericytomas. METHODS: In a review of the Stanford radiosurgery database between 2002 and 2009, the authors found 14 patients who underwent CK therapy for recurrent, metastatic, and residual hemangiopericytomas. A total of 24 tumors were treated and the median patient age was 52 years (range 29-70 years) at the time of initial CK therapy. The median follow-up period was 37 months (10-73 months) and all patients had been previously treated with surgical resection. Mean tumor volume was 9.16 cm3 and the mean marginal and maximum radiosurgical doses to the tumors were 21.2 Gy and 26.8 Gy, respectively. RESULTS: Of the 24 tumors treated, 22 have clinical follow up data at this time. Of those 22 tumors, 12 decreased in size (54.5%), 6 remained unchanged (27.3%), and 4 showed recurrence (18.2%) after CK therapy. Progression-free survival rate was 95%, 71.5%, and 71.5% at 1, 3, and 5 years after multiple CK treatments. The 5-year survival rate after CK was 81%. CONCLUSIONS: CK is an effective and safe management option for hemangiopericytomas. The current series demonstrates a tumor control of 81.8%. Other institutions have demonstrated similar outcomes with stereotactic radiosurgery, with tumor control ranging from 46.4% to 100%. PMID- 21645368 TI - Comparative genomic analysis of Vibrio parahaemolyticus: serotype conversion and virulence. AB - BACKGROUND: Vibrio parahaemolyticus is a common cause of foodborne disease. Beginning in 1996, a more virulent strain having serotype O3:K6 caused major outbreaks in India and other parts of the world, resulting in the emergence of a pandemic. Other serovariants of this strain emerged during its dissemination and together with the original O3:K6 were termed strains of the pandemic clone. Two genomes, one of this virulent strain and one pre-pandemic strain have been sequenced. We sequenced four additional genomes of V. parahaemolyticus in this study that were isolated from different geographical regions and time points. Comparative genomic analyses of six strains of V. parahaemolyticus isolated from Asia and Peru were performed in order to advance knowledge concerning the evolution of V. parahaemolyticus; specifically, the genetic changes contributing to serotype conversion and virulence. Two pre-pandemic strains and three pandemic strains, isolated from different geographical regions, were serotype O3:K6 and either toxin profiles (tdh+, trh-) or (tdh-, trh+). The sixth pandemic strain sequenced in this study was serotype O4:K68. RESULTS: Genomic analyses revealed that the trh+ and tdh+ strains had different types of pathogenicity islands and mobile elements as well as major structural differences between the tdh pathogenicity islands of the pre-pandemic and pandemic strains. In addition, the results of single nucleotide polymorphism (SNP) analysis showed that 94% of the SNPs between O3:K6 and O4:K68 pandemic isolates were within a 141 kb region surrounding the O- and K-antigen-encoding gene clusters. The "core" genes of V. parahaemolyticus were also compared to those of V. cholerae and V. vulnificus, in order to delineate differences between these three pathogenic species. Approximately one-half (49-59%) of each species' core genes were conserved in all three species, and 14-24% of the core genes were species-specific and in different functional categories. CONCLUSIONS: Our data support the idea that the pandemic strains are closely related and that recent South American outbreaks of foodborne disease caused by V. parahaemolyticus are closely linked to outbreaks in India. Serotype conversion from O3:K6 to O4:K68 was likely due to a recombination event involving a region much larger than the O-antigen- and K antigen-encoding gene clusters. Major differences between pathogenicity islands and mobile elements are also likely driving the evolution of V. parahaemolyticus. In addition, our analyses categorized genes that may be useful in differentiating pathogenic Vibrios at the species level. PMID- 21645369 TI - Anti-inflammatory activity of Chios mastic gum is associated with inhibition of TNF-alpha induced oxidative stress. AB - BACKGROUND: Gum of Chios mastic (Pistacia lentiscus var. chia) is a natural antimicrobial agent that has found extensive use in pharmaceutical products and as a nutritional supplement. The molecular mechanisms of its anti-inflammatory activity, however, are not clear. In this work, the potential role of antioxidant activity of Chios mastic gum has been evaluated. METHODS: Scavenging of superoxide radical was investigated by electron spin resonance and spin trapping technique using EMPO spin trap in xanthine oxidase system. Superoxide production in endothelial and smooth muscle cells stimulated with TNF-alpha or angiotensin II and treated with vehicle (DMSO) or mastic gum (0.1-10 MUg/ml) was measured by DHE and HPLC. Cellular H2O2 was measured by Amplex Red. Inhibition of protein kinase C (PKC) with mastic gum was determined by the decrease of purified PKC activity, by inhibition of PKC activity in cellular homogenate and by attenuation of superoxide production in cells treated with PKC activator phorbol 12-myristate 13-acetate (PMA). RESULTS: Spin trapping study did not show significant scavenging of superoxide by mastic gum itself. However, mastic gum inhibited cellular production of superoxide and H2O2 in dose dependent manner in TNF-alpha treated rat aortic smooth muscle cells but did not affect unstimulated cells. TNF alpha significantly increased the cellular superoxide production by NADPH oxidase, while mastic gum completely abolished this stimulation. Mastic gum inhibited the activity of purified PKC, decreased PKC activity in cell homogenate, and attenuated superoxide production in cells stimulated with PKC activator PMA and PKC-dependent angiotensin II in endothelial cells. CONCLUSION: We suggest that mastic gum inhibits PKC which attenuates production of superoxide and H2O2 by NADPH oxidases. This antioxidant property may have direct implication to the anti-inflammatory activity of the Chios mastic gum. PMID- 21645370 TI - Central nervous system exposure of next generation quinoline methanols is reduced relative to mefloquine after intravenous dosing in mice. AB - BACKGROUND: The clinical use of mefloquine (MQ) has declined due to dose-related neurological events. Next generation quinoline methanols (NGQMs) that do not accumulate in the central nervous system (CNS) to the same extent may have utility. In this study, CNS levels of NGQMs relative to MQ were measured and an early lead chemotype was identified for further optimization. EXPERIMENTAL DESIGN: The plasma and brain levels of MQ and twenty five, 4-position modified NGQMs were determined using LCMS/MS at 5 min, 1, 6 and 24 h after IV administration (5 mg/kg) to male FVB mice. Fraction unbound in brain tissue homogenate was assessed in vitro using equilibrium dialysis and this was then used to calculate brain-unbound concentration from the measured brain total concentration. A five-fold reduction CNS levels relative to mefloquine was considered acceptable. Additional pharmacological properties such as permeability and potency were determined. RESULTS: The maximum brain (whole/free) concentrations of MQ were 1807/4.9 ng/g. Maximum whole brain concentrations of NGQMs were 23 - 21546 ng/g. Maximum free brain concentrations were 0.5 to 267 ng/g. Seven (28%) and two (8%) compounds exhibited acceptable whole and free brain concentrations, respectively. Optimization of maximum free brain levels, IC90s (as a measure or potency) and residual plasma concentrations at 24 h (as a surrogate for half-life) in the same molecule may be feasible since they were not correlated. Diamine quinoline methanols were the most promising lead compounds. CONCLUSION: Reduction of CNS levels of NGQMs relative to mefloquine may be feasible. Optimization of this property together with potency and long half-life may be feasible amongst diamine quinoline methanols. PMID- 21645371 TI - Family association study between INSR gene polymorphisms and PCOS in Han Chinese. AB - BACKGROUND: Polycystic ovary syndrome (PCOS) is a complex disease having both genetic and environmental components. Candidate genes with insulin metabolism have been hypothesized to be involved in the etiology of this syndrome. In the present study, we investigated the genetic association between polymorphisms in the insulin receptor (INSR) gene and PCOS. METHODS: A total of 260 family trios were recruited and performed a family-based analysis to assess linkage and association between four single nucleotide polymorphisms (SNPs) (rs1799817, rs2059807, rs8108622 and rs10500204) of INSR gene and PCOS. RESULTS: Using the transmission disequilibrium test (TDT), we failed to find that rs1799817 (p = 0.486), rs2059807 (p = 0.195), rs8108622 (p = 0.866) and rs10500204 (p = 1.0) were significantly overtransmitted to PCOS offspring from their parents. CONCLUSION: No significant evidence of association or linkage was found in the four tested markers, indicating that our family samples did not support susceptibility of the INSR gene to PCOS. PMID- 21645372 TI - Combined low initial DNA damage and high radiation-induced apoptosis confers clinical resistance to long-term toxicity in breast cancer patients treated with high-dose radiotherapy. AB - BACKGROUND: Either higher levels of initial DNA damage or lower levels of radiation-induced apoptosis in peripheral blood lymphocytes have been associated to increased risk for develop late radiation-induced toxicity. It has been recently published that these two predictive tests are inversely related. The aim of the present study was to investigate the combined role of both tests in relation to clinical radiation-induced toxicity in a set of breast cancer patients treated with high dose hyperfractionated radical radiotherapy. METHODS: Peripheral blood lymphocytes were taken from 26 consecutive patients with locally advanced breast carcinoma treated with high-dose hyperfractioned radical radiotherapy. Acute and late cutaneous and subcutaneous toxicity was evaluated using the Radiation Therapy Oncology Group morbidity scoring schema. The mean follow-up of survivors (n = 13) was 197.23 months. Radiosensitivity of lymphocytes was quantified as the initial number of DNA double-strand breaks induced per Gy and per DNA unit (200 Mbp). Radiation-induced apoptosis (RIA) at 1, 2 and 8 Gy was measured by flow cytometry using annexin V/propidium iodide. RESULTS: Mean DSB/Gy/DNA unit obtained was 1.70 +/- 0.83 (range 0.63-4.08; median, 1.46). Radiation-induced apoptosis increased with radiation dose (median 12.36, 17.79 and 24.83 for 1, 2, and 8 Gy respectively). We observed that those "expected resistant patients" (DSB values lower than 1.78 DSB/Gy per 200 Mbp and RIA values over 9.58, 14.40 or 24.83 for 1, 2 and 8 Gy respectively) were at low risk of suffer severe subcutaneous late toxicity (HR 0.223, 95%CI 0.073-0.678, P = 0.008; HR 0.206, 95%CI 0.063-0.677, P = 0.009; HR 0.239, 95%CI 0.062-0.929, P = 0.039, for RIA at 1, 2 and 8 Gy respectively) in multivariate analysis. CONCLUSIONS: A radiation-resistant profile is proposed, where those patients who presented lower levels of initial DNA damage and higher levels of radiation induced apoptosis were at low risk of suffer severe subcutaneous late toxicity after clinical treatment at high radiation doses in our series. However, due to the small sample size, other prospective studies with higher number of patients are needed to validate these results. PMID- 21645373 TI - View the label before you view the movie: a field experiment into the impact of portion size and Guideline Daily Amounts labelling on soft drinks in cinemas. AB - BACKGROUND: Large soft drink sizes increase consumption, and thereby contribute to obesity. Portion size labelling may help consumers to select more appropriate food portions. This study aimed to assess the effectiveness of portion size and caloric Guidelines for Daily Amounts (GDA) labelling on consumers' portion size choices and consumption of regular soft drinks. METHODS: A field experiment that took place on two subsequent evenings in a Dutch cinema. Participants (n = 101) were asked to select one of five different portion sizes of a soft drink. Consumers were provided with either portion size and caloric GDA labelling (experimental condition) or with millilitre information (control condition). RESULTS: Labelling neither stimulated participants to choose small portion sizes (OR = .75, p = .61, CI: .25 - 2.25), nor did labelling dissuade participants to choose large portion sizes (OR = .51, p = .36, CI: .12 - 2.15). CONCLUSIONS: Portion size and caloric GDA labelling were found to have no effect on soft drink intake. Further research among a larger group of participants combined with pricing strategies is required. The results of this study are relevant for the current public health debate on food labelling. PMID- 21645374 TI - Do mitochondria play a role in remodelling lace plant leaves during programmed cell death? AB - BACKGROUND: Programmed cell death (PCD) is the regulated death of cells within an organism. The lace plant (Aponogeton madagascariensis) produces perforations in its leaves through PCD. The leaves of the plant consist of a latticework of longitudinal and transverse veins enclosing areoles. PCD occurs in the cells at the center of these areoles and progresses outwards, stopping approximately five cells from the vasculature. The role of mitochondria during PCD has been recognized in animals; however, it has been less studied during PCD in plants. RESULTS: The following paper elucidates the role of mitochondrial dynamics during developmentally regulated PCD in vivo in A. madagascariensis. A single areole within a window stage leaf (PCD is occurring) was divided into three areas based on the progression of PCD; cells that will not undergo PCD (NPCD), cells in early stages of PCD (EPCD), and cells in late stages of PCD (LPCD). Window stage leaves were stained with the mitochondrial dye MitoTracker Red CMXRos and examined. Mitochondrial dynamics were delineated into four categories (M1-M4) based on characteristics including distribution, motility, and membrane potential (DeltaPsim). A TUNEL assay showed fragmented nDNA in a gradient over these mitochondrial stages. Chloroplasts and transvacuolar strands were also examined using live cell imaging. The possible importance of mitochondrial permeability transition pore (PTP) formation during PCD was indirectly examined via in vivo cyclosporine A (CsA) treatment. This treatment resulted in lace plant leaves with a significantly lower number of perforations compared to controls, and that displayed mitochondrial dynamics similar to that of non-PCD cells. CONCLUSIONS: Results depicted mitochondrial dynamics in vivo as PCD progresses within the lace plant, and highlight the correlation of this organelle with other organelles during developmental PCD. To the best of our knowledge, this is the first report of mitochondria and chloroplasts moving on transvacuolar strands to form a ring structure surrounding the nucleus during developmental PCD. Also, for the first time, we have shown the feasibility for the use of CsA in a whole plant system. Overall, our findings implicate the mitochondria as playing a critical and early role in developmentally regulated PCD in the lace plant. PMID- 21645375 TI - Experimental mutation-accumulation on the X chromosome of Drosophila melanogaster reveals stronger selection on males than females. AB - BACKGROUND: Sex differences in the magnitude or direction of mutational effect may be important to a variety of population processes, shaping the mutation load and affecting the cost of sex itself. These differences are expected to be greatest after sexual maturity. Mutation-accumulation (MA) experiments provide the most direct way to examine the consequences of new mutations, but most studies have focused on juvenile viability without regard to sex, and on autosomes rather than sex chromosomes; both adult fitness and X-linkage have been little studied. We therefore investigated the effects of 50 generations of X chromosome mutation accumulation on the fitness of males and females derived from an outbred population of Drosophila melanogaster. RESULTS: Fitness declined rapidly in both sexes as a result of MA, but adult males showed markedly greater fitness loss relative to their controls compared to females expressing identical genotypes, even when females were made homozygous for the X. We estimate that these mutations are partially additive (h ~ 0.3) in females. In addition, the majority of new mutations appear to harm both males and females. CONCLUSIONS: Our data helps fill a gap in our understanding of the consequences of sexual selection for genetic load, and suggests that stronger selection on males may indeed purge deleterious mutations affecting female fitness. PMID- 21645376 TI - Low avidity of human papillomavirus (HPV) type 16 antibodies is associated with increased risk of low-risk but not high-risk HPV type prevalence. AB - BACKGROUND: Low avidity of antibodies against viral, bacterial and parasitic agents has been used for differential diagnosis of acute versus recent/past infections. The low-avidity antibodies may however, persist for a longer period in some individuals. FINDINGS: We studied the association of human papillomavirus (HPV) type 16 antibody avidity with seroprevalence to HPV types 6/11/18/31/33/45. Antibody avidity was analysed for 365 HPV16 seropositive pregnant Finnish and Ugandan women using a modified ELISA.Low avidity of HPV16 antibodies was found in 15% of Finnish and 26% of Ugandan women. Ugandan women with low-avidity HPV16 antibodies had an increased risk estimate for HPV6/11 (odds ratio, OR 2.9; 95%CI 1.01-8.4) seropositivity but not to high-risk HPV types 18/31/33/45. CONCLUSION: Association of the low avidity HPV16 antibody "phenotype" with possible susceptibility to infections with other HPV types warrants investigation. PMID- 21645377 TI - CloudAligner: A fast and full-featured MapReduce based tool for sequence mapping. AB - BACKGROUND: Research in genetics has developed rapidly recently due to the aid of next generation sequencing (NGS). However, massively-parallel NGS produces enormous amounts of data, which leads to storage, compatibility, scalability, and performance issues. The Cloud Computing and MapReduce framework, which utilizes hundreds or thousands of shared computers to map sequencing reads quickly and efficiently to reference genome sequences, appears to be a very promising solution for these issues. Consequently, it has been adopted by many organizations recently, and the initial results are very promising. However, since these are only initial steps toward this trend, the developed software does not provide adequate primary functions like bisulfite, pair-end mapping, etc., in on-site software such as RMAP or BS Seeker. In addition, existing MapReduce-based applications were not designed to process the long reads produced by the most recent second-generation and third-generation NGS instruments and, therefore, are inefficient. Last, it is difficult for a majority of biologists untrained in programming skills to use these tools because most were developed on Linux with a command line interface. RESULTS: To urge the trend of using Cloud technologies in genomics and prepare for advances in second- and third-generation DNA sequencing, we have built a Hadoop MapReduce-based application, CloudAligner, which achieves higher performance, covers most primary features, is more accurate, and has a user-friendly interface. It was also designed to be able to deal with long sequences. The performance gain of CloudAligner over Cloud-based counterparts (35 to 80%) mainly comes from the omission of the reduce phase. In comparison to local-based approaches, the performance gain of CloudAligner is from the partition and parallel processing of the huge reference genome as well as the reads. The source code of CloudAligner is available at http://cloudaligner.sourceforge.net/ and its web version is at http://mine.cs.wayne.edu:8080/CloudAligner/. CONCLUSIONS: Our results show that CloudAligner is faster than CloudBurst, provides more accurate results than RMAP, and supports various input as well as output formats. In addition, with the web based interface, it is easier to use than its counterparts. PMID- 21645378 TI - Characteristics and predictors of readiness to quit among emergency medical patients presenting with respiratory symptoms. AB - PURPOSE: To examine behavioral factors that lead patients to consider quitting smoking and features associated with readiness to quit among adults who are seeking treatment in the emergency department (ED) for respiratory symptoms. METHODS: A toal of 665 adult smokers seeking treatment in an ED for respiratory symptoms and respiratory illness answered survey questions during the ED visit. RESULTS: Patients self-reported "readiness to quit" was broadly distributed among this patient population. Patients with COPD, pneumonia or asthma perceived higher risks from smoking than other patients with respiratory complaints. Over half of all participants had scores indicative of depression. Regression analysis showed that prior efforts to quit, confidence, perceived importance of quitting and decisional balance were each significantly predictive of readiness to quit, accounting for 40% of the variance. CONCLUSIONS: While many of these patients appear unaware of the connection between their symptoms and their smoking, patients with diagnosed chronic respiratory illness perceived higher risks from their smoking. In patients who do not perceive these risks, physician intervention may increase perceived risk from smoking and perceived importance of quitting. Interventions designed for the ED setting targeting this patient population should consider screening for depressive symptoms and, when appropriate, making referrals for further evaluation and/or treatment. Medications that can help alleviate depression and withdrawal symptoms while quitting smoking, such as bupropion, may be particularly useful for this subset of patients, as depression is a substantial barrier to quitting. PMID- 21645379 TI - Using quantile regression to investigate racial disparities in medication non adherence. AB - BACKGROUND: Many studies have investigated racial/ethnic disparities in medication non-adherence in patients with type 2 diabetes using common measures such as medication possession ratio (MPR) or gaps between refills. All these measures including MPR are quasi-continuous and bounded and their distribution is usually skewed. Analysis of such measures using traditional regression methods that model mean changes in the dependent variable may fail to provide a full picture about differential patterns in non-adherence between groups. METHODS: A retrospective cohort of 11,272 veterans with type 2 diabetes was assembled from Veterans Administration datasets from April 1996 to May 2006. The main outcome measure was MPR with quantile cutoffs Q1-Q4 taking values of 0.4, 0.6, 0.8 and 0.9. Quantile-regression (QReg) was used to model the association between MPR and race/ethnicity after adjusting for covariates. Comparison was made with commonly used ordinary-least-squares (OLS) and generalized linear mixed models (GLMM). RESULTS: Quantile-regression showed that Non-Hispanic-Black (NHB) had statistically significantly lower MPR compared to Non-Hispanic-White (NHW) holding all other variables constant across all quantiles with estimates and p values given as -3.4% (p = 0.11), -5.4% (p = 0.01), -3.1% (p = 0.001), and -2.00% (p = 0.001) for Q1 to Q4, respectively. Other racial/ethnic groups had lower adherence than NHW only in the lowest quantile (Q1) of about -6.3% (p = 0.003). In contrast, OLS and GLMM only showed differences in mean MPR between NHB and NHW while the mean MPR difference between other racial groups and NHW was not significant. CONCLUSION: Quantile regression is recommended for analysis of data that are heterogeneous such that the tails and the central location of the conditional distributions vary differently with the covariates. QReg provides a comprehensive view of the relationships between independent and dependent variables (i.e. not just centrally but also in the tails of the conditional distribution of the dependent variable). Indeed, without performing QReg at different quantiles, an investigator would have no way of assessing whether a difference in these relationships might exist. PMID- 21645380 TI - Variable antibiotic susceptibility patterns among Streptomyces species causing actinomycetoma in man and animals. AB - BACKGROUND: Drug therapy is recommended in conjunction with surgery in treatment of actinomycetoma. The specific prescription depends on the type of bacteria (actinomycetoma) or fungi (eumycetoma) causing the disease and their in vitro antimicrobial susceptibility. OBJECTIVES: To investigate the antimicrobial susceptibility among isolates of Streptomyces spp. isolated from cases of actinomycetoma in man and animals in Sudan. METHODS: Streptomyces strains (n = 18) isolated from cases of actinomycetoma were tested in vitro against 15 commonly prescribed antibacterial agents using MIC agar dilution method as per standard guidelines. RESULTS: Streptomyces strains isolated from actinomycetoma fall into various phenotypic groups. All of the strains were inhibited by novobiocin (8 MUg/mL), gentamycin (8, 32 MUg/mL) and doxycycline (32 MUg/mL). Fusidic acid (64 MUg/mL) inhibited 94.4% of the strains; bacitracin, streptomycin, cephaloridine, clindamycin, ampicillin, rifampicin and tetracycline (64 MUg/mL) inhibited between 61.1 and 77.8% of the strains. All strains were found resistant to amphotericin B (64 MUg/mL), penicillin (20 MUg/mL) and sulphamethoxazole (64 MUg/mL). CONCLUSIONS: Saprophytic Streptomyces spp. cause actinomycetoma in man and animal belong to separate phenotypes and have a wide range of susceptibility patterns to antimicrobial agents, which pose a lot of difficulties in selecting effective in vivo treatment for actinomycetoma. PMID- 21645381 TI - Community-based knowledge translation: unexplored opportunities. AB - BACKGROUND: Knowledge translation is an interactive process of knowledge exchange between health researchers and knowledge users. Given that the health system is broad in scope, it is important to reflect on how definitions and applications of knowledge translation might differ by setting and focus. Community-based organizations and their practitioners share common characteristics related to their setting, the evidence used in this setting, and anticipated outcomes that are not, in our experience, satisfactorily reflected in current knowledge translation approaches, frameworks, or tools. DISCUSSION: Community-based organizations face a distinctive set of challenges and concerns related to engaging in the knowledge translation process, suggesting a unique perspective on knowledge translation in these settings. Specifically, community-based organizations tend to value the process of working in collaboration with multi sector stakeholders in order to achieve an outcome. A feature of such community based collaborations is the way in which 'evidence' is conceptualized or defined by these partners, which may in turn influence the degree to which generalizable research evidence in particular is relevant and useful when balanced against more contextually-informed knowledge, such as tacit knowledge. Related to the issues of evidence and context is the desire for local information. For knowledge translation researchers, developing processes to assist community-based organizations to adapt research findings to local circumstances may be the most helpful way to advance decision making in this area. A final characteristic shared by community-based organizations is involvement in advocacy activities, a function that has been virtually ignored in traditional knowledge translation approaches. SUMMARY: This commentary is intended to stimulate further discussion in the area of community-based knowledge translation. Knowledge translation, and exchange, between communities, community-based organizations, decision makers, and researchers is likely to be beneficial when ensuring that 'evidence' meets the needs of all end users and that decisions are based on both relevant research and community requirements. Further exploratory work is needed to identify alternative methods for evaluating these strategies when applied within community based settings. PMID- 21645382 TI - A note on the use of the generalized odds ratio in meta-analysis of association studies involving bi- and tri-allelic polymorphisms. AB - BACKGROUND: The generalized odds ratio (GOR) was recently suggested as a genetic model-free measure for association studies. However, its properties were not extensively investigated. We used Monte Carlo simulations to investigate type-I error rates, power and bias in both effect size and between-study variance estimates of meta-analyses using the GOR as a summary effect, and compared these results to those obtained by usual approaches of model specification. We further applied the GOR in a real meta-analysis of three genome-wide association studies in Alzheimer's disease. FINDINGS: For bi-allelic polymorphisms, the GOR performs virtually identical to a standard multiplicative model of analysis (e.g. per allele odds ratio) for variants acting multiplicatively, but augments slightly the power to detect variants with a dominant mode of action, while reducing the probability to detect recessive variants. Although there were differences among the GOR and usual approaches in terms of bias and type-I error rates, both simulation- and real data-based results provided little indication that these differences will be substantial in practice for meta-analyses involving bi allelic polymorphisms. However, the use of the GOR may be slightly more powerful for the synthesis of data from tri-allelic variants, particularly when susceptibility alleles are less common in the populations (<=10%). This gain in power may depend on knowledge of the direction of the effects. CONCLUSIONS: For the synthesis of data from bi-allelic variants, the GOR may be regarded as a multiplicative-like model of analysis. The use of the GOR may be slightly more powerful in the tri-allelic case, particularly when susceptibility alleles are less common in the populations. PMID- 21645383 TI - The roles of herbal remedies in survival and quality of life among long-term breast cancer survivors--results of a prospective study. AB - BACKGROUND: Few data exist on survival or health-related quality of life (QOL) related to herbal remedy use among long-term breast cancer survivors. The objective of this report is to examine whether herbal remedy use is associated with survival or the health-related QOL of these long-term breast cancer survivors. METHODS: In 1999-2000, we collected the information of herbal remedy use and QOL during a telephone interview with 371 Los Angeles Non Hispanic/Hispanic white women who had survived more than 10 years after breast cancer diagnosis. QOL was measured using the Medical Outcomes Study Short Form-36 (SF-36) questionnaire. Patients were followed for mortality from the baseline interview through 2007. 299 surviving patients completed a second telephone interview on QOL in 2002-2004. We used multivariable Cox proportional hazards methods to estimate relative risks (RR) and 95% confidence intervals (CI) for mortality and applied multivariable linear regression models to compare average SF-36 change scores (follow-up - baseline) between herbal remedy users and non users. RESULTS: Fifty-nine percent of participants were herbal remedy users at baseline. The most commonly used herbal remedies were echinacea, herbal teas, and ginko biloba. Herbal remedy use was associated with non-statistically significant increases in the risks for all-cause (44 deaths, RR=1.28, 95% CI=0.62-2.64) and breast cancer (33 deaths, RR=1.78, 95% CI=0.72-4.40) mortality. Both herbal remedy users' and non-users' mental component summary scores on the SF-36 increased similarly from the first survey to the second survey (P=0.16), but herbal remedy users' physical component summary scores decreased more than those of non-users (-5.7 vs. -3.2, P=0.02). CONCLUSIONS: Our data provide some evidence that herbal remedy use is associated with poorer survival and a poorer physical component score for health-related QOL among women who have survived breast cancer for at least 10 years. These conclusions are based on exploratory analyses of data from a prospective study using two-sided statistical tests with no correction for multiple testing and are limited by few deaths for mortality analysis and lack of information on when herbal remedy use was initiated or duration of or reasons for use. PMID- 21645384 TI - Relation between mean arterial pressure and renal function in the early phase of shock: a prospective, explorative cohort study. AB - INTRODUCTION: Because of disturbed renal autoregulation, patients experiencing hypotension-induced renal insult might need higher levels of mean arterial pressure (MAP) than the 65 mmHg recommended level in order to avoid the progression of acute kidney insufficiency (AKI). METHODS: In 217 patients with sustained hypotension, enrolled and followed prospectively, we compared the evolution of the mean arterial pressure (MAP) during the first 24 hours between patients who will show AKI 72 hours after inclusion (AKIh72) and patients who will not. AKIh72 was defined as the need of renal replacement therapy or "Injury" or "Failure" classes of the 5-stage RIFLE classification (Risk, Injury, Failure, Loss of kidney function, End-stage renal disease) for acute kidney insufficiency using the creatinine and urine output criteria. This comparison was performed in four different subgroups of patients according to the presence or not of AKI at the sixth hour after inclusion (AKIh6 as defined as a serum creatinine level above 1.5 times baseline value within the first six hours) and the presence or not of septic shock at inclusion.The ability of MAP averaged over H6 to H24 to predict AKIh72 was assessed by the area under the receiver operating characteristic curve (AUC) and compared between groups. RESULTS: The MAP averaged over H6 to H24 or over H12 to H24 was significantly lower in patients who showed AKIh72 than in those who did not, only in septic shock patients with AKIh6, whereas no link was found between MAP and AKIh72 in the three others subgroups of patients. In patients with septic shock plus AKIh6, MAP averaged over H6 to H24 or over H12 to H24 had an AUC of 0.83 (0.72 to 0.92) or 0.84 (0.72 to 0.92), respectively, to predict AKIh72 . In these patients, the best level of MAP to prevent AKIh72 was between 72 and 82 mmHg. CONCLUSIONS: MAP about 72 to 82 mmHg could be necessary to avoid acute kidney insufficiency in patients with septic shock and initial renal function impairment. PMID- 21645385 TI - The contribution of aestivating mosquitoes to the persistence of Anopheles gambiae in the Sahel. AB - BACKGROUND: Persistence of African anophelines throughout the long dry season (4 8 months) when no surface waters are available remains one of the enduring mysteries of medical entomology. Recent studies demonstrated that aestivation (summer diapause) is one mechanism that allows the African malaria mosquito, Anopheles gambiae, to persist in the Sahel. However, migration from distant localities - where reproduction continues year-round - might also be involved. METHODS: To assess the contribution of aestivating adults to the buildup of populations in the subsequent wet season, two villages subjected to weekly pyrethrum sprays throughout the dry season were compared with two nearby villages, which were only monitored. If aestivating adults are the main source of the subsequent wet-season population, then the subsequent wet-season density in the treated villages will be lower than in the control villages. Moreover, since virtually only M-form An. gambiae are found during the dry season, the reduction should be specific to the M form, whereas no such difference is predicted for S form An. gambiae or Anopheles arabiensis. On the other hand, if migrants arriving with the first rain are the main source, no differences between treated and control villages are expected across all members of the An. gambiae complex. RESULTS: The wet-season density of the M form in treated villages was 30% lower than that in the control (P < 10-4, permutation test), whereas no significant differences were detected in the S form or An. arabiensis. CONCLUSIONS: These results support the hypothesis that the M form persist in the arid Sahel primarily by aestivation, whereas the S form and An. arabiensis rely on migration from distant locations. Implications for malaria control are discussed. PMID- 21645386 TI - Improved mapping strategy to better inform policy on the control of schistosomiasis and soil-transmitted helminthiasis in Sierra Leone. AB - BACKGROUND: Schistosomiasis and soil-transmitted helminthiasis (STH) are endemic in Sierra Leone confirmed by national mapping in 2008. To better inform planning of preventive chemotherapy strategy, another survey was conducted before mass drug administration (MDA) in seven districts according to the mapping results or local knowledge. Fifty-nine chiefdoms and one school in every chiefdom were selected. Thirty school children aged 9-14 years from each school (total: 1760) were examined by parasitological methods for infection with Schistosoma mansoni and STHs. RESULTS: The overall prevalence of S. mansoni was 40.2% (95% confidence interval (CI): 37.9-42.5%), particularly in Kailahun (63.3%), Kenema (46.7%), Koinadugu (41.9%) and Kono (71.7%). The results demonstrated the focal distribution of S. mansoni in Bo, Tonkolili and Bombali districts with prevalence ranging from 0.0-63.3%, 3.3-90.0% and 0.0-67.9% respectively. The arithmetic mean intensity of S. mansoni infection was 95.4 epg (95% CI: 61.4-129.5 epg), Heavy mean intensity of infection was found in Kailahun (120.2 epg), Kenema (104.5 epg), Koinadugu (112.3 epg) and Kono (250.3 epg). Heavy or moderate infection with S. mansoni occurred in 20.7% of children examined. Hookworm prevalence was moderate: 31.2% (95% CI: 29.1-33.4%), but high in Bo (50.0%) and Tonkolili (56.7%). Hookworm intensity of infection was light with a mean epg of 53.0 (95% CI: 38.4-67.7 epg). Prevalence and intensity of Ascaris lumbricoides (1.5%, 17.8 epg) and Trichuris trichiura (2.5%, 20.3 epg) was low. CONCLUSIONS: The prediction by previous spatial analysis that S. mansoni was highly endemic across north-eastern Sierra Leone was confirmed with a significant proportion of children heavily or moderately infected. The distribution of S. mansoni in Bo, Tonkolili and Bombali districts ranged widely, highlighting the importance of considering the nature of focal transmission in national mapping exercises. These results were used to refine the MDA for schistosomiasis control to chiefdom implementation units rather than the entire district in these 3 districts. The survey demonstrated that sufficient number of survey sites for schistosomiasis mapping in each district should be used to provide a better national planning of MDA activities, and that it is affordable with the contributions from all parties involved and national resources mobilized. PMID- 21645387 TI - Meningitis in adult patients with a negative direct cerebrospinal fluid examination: value of cytochemical markers for differential diagnosis. AB - INTRODUCTION: The objective of this study was to determine the ability of various parameters commonly used for the diagnosis of acute meningitis to differentiate between bacterial and viral meningitis, in adult patients with a negative direct cerebrospinal fluid (CSF) examination. METHODS: This was a prospective study, started in 1997, including all patients admitted to the emergency unit with acute meningitis and a negative direct CSF examination. Serum and CSF samples were taken immediately on admission. The patients were divided into two groups according to the type of meningitis: bacterial (BM; group I) or viral (VM; group II). The CSF parameters investigated were cytology, protein, glucose, and lactate; the serum parameters evaluated were C-reactive protein and procalcitonin. CSF/serum glucose and lactate ratios were also assessed. RESULTS: Of the 254 patients with meningitis with a negative direct CSF examination, 35 had BM and 181, VM. The most highly discriminative parameters for the differential diagnosis of BM proved to be CSF lactate, with a sensitivity of 94%, a specificity of 92%, a negative predictive value of 99%, a positive predictive value of 82% at a diagnostic cut-off level of 3.8 mmol/L (area under the curve (AUC), 0.96; 95% confidence interval (CI), 0.95 to 1), and serum procalcitonin, with a sensitivity of 95%, a specificity of 100%, a negative predictive value of 100%, and a positive predictive value of 97% at a diagnostic cut-off level of 0.28 ng/ml (AUC, 0.99; 95% CI, 0.99 to 1). CONCLUSIONS: Serum procalcitonin and CSF lactate concentrations appear to be the most highly discriminative parameters for the differential diagnosis of BM and VM. PMID- 21645388 TI - Patient acceptance of universal screening for hepatitis C virus infection. AB - BACKGROUND: In the United States, about 70% of 2.9-3.7 million people with hepatitis C (HCV) are unaware of their infection. Although universal screening might be a cost-effective way to identify infections, prevent morbidity, and reduce transmission, few efforts have been made to determine patient opinions about new approaches to screening. METHODS: We surveyed 200 patients in August 2010 at five outpatient clinics of a major public urban medical center in Seattle, WA, with an 85.8% response rate. RESULTS: The sample was 55.3% women, median 47 years of age, and 56.3% white and 32.7% African or African-American; 9.5% and 2.5% reported testing positive for HCV and HIV, respectively. The vast majority of patients supported universal screening for HCV. When presented with three options for screening, 48% preferred universal testing without being informed that they were being tested or provided with negative results, 37% preferred testing with the chance to "opt-out" of being tested and without being provided with negative results, and 15% preferred testing based on clinician judgment. Results were similar for HIV screening. CONCLUSIONS: Patients support universal screening for HCV, even if that screening involves testing without prior consent or the routine provision of negative test results. Current screening guidelines and procedures should be reconsidered in light of patient priorities. PMID- 21645389 TI - Severe forms of partial androgen insensitivity syndrome due to p.L830F novel mutation in androgen receptor gene in a Brazilian family. AB - BACKGROUND: The androgen insensitivity syndrome may cause developmental failure of normal male external genitalia in individuals with 46,XY karyotype. It results from the diminished or absent biological action of androgens, which is mediated by the androgen receptor in both embryo and secondary sex development. Mutations in the androgen receptor gene, located on the X chromosome, are responsible for the disease. Almost 70% of 46,XY affected individuals inherited mutations from their carrier mothers. FINDINGS: Molecular abnormalities in the androgen receptor gene in individuals of a Brazilian family with clinical features of severe forms of partial androgen insensitivity syndrome were evaluated. Seven members (five 46,XY females and two healthy mothers) of the family were included in the investigation. The coding exons and exon-intron junctions of androgen receptor gene were sequenced. Five 46,XY members of the family have been found to be hemizygous for the c.3015C>T nucleotide change in exon 7 of the androgen receptor gene, whereas the two 46,XX mothers were heterozygote carriers. This nucleotide substitution leads to the p.L830F mutation in the androgen receptor. CONCLUSIONS: The novel p.L830F mutation is responsible for grades 5 and 6 of partial androgen insensitivity syndrome in two generations of a Brazilian family. PMID- 21645390 TI - A comparison of dose-response characteristics of four NTCP models using outcomes of radiation-induced optic neuropathy and retinopathy. AB - BACKGROUND: Biological models are used to relate the outcome of radiation therapy to dose distribution. As use of biological models in treatment planning expands, uncertainties associated with the use of specific models for predicting outcomes should be understood and quantified. In particular, the question to what extent model predictions are data-driven or dependent on the choice of the model has to be explored. METHODS: Four dose-response models--logistic, log-logistic, Poisson based and probit--were tested for their ability and consistency in describing dose-response data for radiation-induced optic neuropathy (RION) and retinopathy (RIRP). Dose to the optic nerves was specified as the minimum dose, Dmin, received by any segment of the organ to which the damage was diagnosed by ophthalmologic evaluation. For retinopathy, the dose to the retina was specified as the highest isodose covering at least 1/3 of the retinal surface (D33%) that geometrically covered the observed retinal damage. Data on both complications were modeled separately for patients treated once daily and twice daily. Model parameters D50 and gamma and corresponding confidence intervals were obtained using maximum-likelihood method. RESULTS: Model parameters were reasonably consistent for RION data for patients treated once daily, D50 ranging from 94.2 to 104.7 Gy and gamma from 0.88 to 1.41. Similar consistency was seen for RIRP data which span a broad range of complication incidence, with D50 from 72.2 to 75.0 Gy and gamma from 1.51 to 2.16 for patients treated twice daily; 72.2-74.0 Gy and 0.84-1.20 for patients treated once daily. However, large variations were observed for RION in patients treated twice daily, D50 from 96.3 to 125.2 Gy and gamma from 0.80 to 1.56. Complication incidence in this dataset in any dose group did not exceed 20%. CONCLUSIONS: For the considered data sets, the log-logistic model tends to lead to larger D50 and lower gamma compared to other models for all datasets. Statements regarding normal tissue radiosensitivity and steepness of dose-response, based on model parameters, should be made with caution as the latter are not only model-dependent but also sensitive to the range of complication incidence exhibited by clinical data. PMID- 21645391 TI - Tau oligomers impair memory and induce synaptic and mitochondrial dysfunction in wild-type mice. AB - BACKGROUND: The correlation between neurofibrillary tangles of tau and disease progression in the brains of Alzheimer's disease (AD) patients remains an area of contention. Innovative data are emerging from biochemical, cell-based and transgenic mouse studies that suggest that tau oligomers, a pre-filament form of tau, may be the most toxic and pathologically significant tau aggregate. RESULTS: Here we report that oligomers of recombinant full-length human tau protein are neurotoxic in vivo after subcortical stereotaxic injection into mice. Tau oligomers impaired memory consolidation, whereas tau fibrils and monomers did not. Additionally, tau oligomers induced synaptic dysfunction by reducing the levels of synaptic vesicle-associated proteins synaptophysin and septin-11. Tau oligomers produced mitochondrial dysfunction by decreasing the levels of NADH ubiquinone oxidoreductase (electron transport chain complex I), and activated caspase-9, which is related to the apoptotic mitochondrial pathway. CONCLUSIONS: This study identifies tau oligomers as an acutely toxic tau species in vivo, and suggests that tau oligomers induce neurodegeneration by affecting mitochondrial and synaptic function, both of which are early hallmarks in AD and other tauopathies. These results open new avenues for neuroprotective intervention strategies of tauopathies by targeting tau oligomers. PMID- 21645392 TI - Prevalence of physical inactivity and barriers to physical activity among obese attendants at a community health-care center in Karachi, Pakistan. AB - BACKGROUND: Overweight and obesity are significant public health problems worldwide with serious health consequences. With increasing urbanization and modernization there has been an increase in prevalence of obesity that is attributed to reduced levels of physical activity (PA). However, little is known about the prevalence of physical inactivity and factors that prohibit physical activity among Pakistani population. This cross-sectional study is aimed at estimating the prevalence of physical inactivity, and determining associated barriers in obese attendants accompanying patients coming to a Community Health Center in Karachi, Pakistan. FINDINGS: PA was assessed by using international physical activity questionnaire (IPAQ). Barriers to PA were also assessed in inactive obese attendants. A pre-tested questionnaire was used to collect data from a total of 350 obese attendants. Among 350 study participants 254 (72.6%) were found to be physically inactive (95% CI: 68.0%, 77.2%). Multivariable logistic regression analysis indicated that age greater than 33 years, BMI greater than 33 kg/m2 and family history of obesity were independently and significantly associated with physical inactivity. Moreover, there was a significant interaction between family structure and gender; females living in extended families were about twice more likely to be inactive, whereas males from extended families were six times more likely to be inactive relative to females from nuclear families. Lack of information, motivation and skills, spouse & family support, accessibility to places for physical activity, cost effective facilities and time were found to be important barriers to PA. CONCLUSIONS: Considering the public health implications of physical inactivity it is essential to promote PA in context of an individual's health and environment. Findings highlight considerable barriers to PA among obese individuals that need to be addressed during counseling sessions with physicians. PMID- 21645393 TI - What do we know about the non-work determinants of workers' mental health? A systematic review of longitudinal studies. AB - BACKGROUND: In the past years, cumulative evidence has convincingly demonstrated that the work environment is a critical determinant of workers' mental health. Nevertheless, much less attention has been dedicated towards understanding the pathways through which other pivotal life environments might also concomitantly intervene, along with the work environment, to bring about mental health outcomes in the workforce. The aim of this study consisted in conducting a systematic review examining the relative contribution of non-work determinants to the prediction of workers' mental health in order to bridge that gap in knowledge. METHODS: We searched electronic databases and bibliographies up to 2008 for observational longitudinal studies jointly investigating work and non-work determinants of workers' mental health. A narrative synthesis (MOOSE) was performed to synthesize data and provide an assessment of study conceptual and methodological quality. RESULTS: Thirteen studies were selected for evaluation. Seven of these were of relatively high methodological quality. Assessment of study conceptual quality yielded modest analytical breadth and depth in the ways studies conceptualized the non-work domain as defined by family, network and community/society-level indicators. We found evidence of moderate strength supporting a causal association between social support from the networks and workers' mental health, but insufficient evidence of specific indicator involvement for other analytical levels considered (i.e., family, community/society). CONCLUSIONS: Largely underinvestigated, non-work determinants are important to the prediction of workers' mental health. More longitudinal studies concomitantly investigating work and non-work determinants of workers' mental health are warranted to better inform healthy workplace research, intervention, and policy. PMID- 21645394 TI - Comparison of early-, late-, and non-participants in a school-based asthma management program for urban high school students. AB - BACKGROUND: To assess bias and generalizability of results in randomized controlled trials (RCT), investigators compare participants to non-participants or early- to late-participants. Comparisons can also inform the recruitment approach, especially when working with challenging populations, such as urban adolescents. In this paper, we describe characteristics by participant status of urban teens eligible to participate in a RCT of a school-based, web-based asthma management program. METHODS: The denominator for this analysis was all students found to be eligible to participate in the RCT. Data were analyzed for participants and non-participants of the RCT, as well as for students that enrolled during the initially scheduled recruitment period (early-participants) and persons that delayed enrollment until the following fall when recruitment was re-opened to increase sample size (late-participants). Full Time Equivalents (FTEs) of staff associated with recruitment were estimated. RESULTS: Of 1668 teens eligible for the RCT, 386 enrolled early, and 36 enrolled late, leaving 1246 non-participants. Participants were younger (p < 0.01), more likely to be diagnosed, use asthma medication, and have moderate-to-severe disease than non participants, odds ratios (95% Confidence Intervals) = 2.1(1.7-2.8), 1.7(1.3 2.1), 1.4(1.0-1.8), respectively. ORs were elevated for the association of late participation with Medicaid enrollment, 1.9(0.7-5.1) and extrinsic motivation to enroll, 1.7(0.6-5.0). Late-participation was inversely related to study compliance for teens and caregivers, ORs ranging from 0.1 to 0.3 (all p-values < 0.01). Early- and late-participants required 0.45 FTEs/100 and 3.3 FTEs/100, respectively. CONCLUSIONS: Recruitment messages attracted youth with moderate-to severe asthma, but extending enrollment was costly, resulting in potentially less motivated, and certainly less compliant, participants. Investigators must balance internal versus external validity in the decision to extend recruitment. Gains in sample size and external validity may be offset by the cost of additional staff time and the threat to internal validity caused by lower participant follow-up. TRIAL REGISTRATION: ClinicalTrials.gov: NCT00201058. PMID- 21645395 TI - Does thermoregulatory behavior maximize reproductive fitness of natural isolates of Caenorhabditis elegans? AB - BACKGROUND: A central premise of physiological ecology is that an animal's preferred body temperature should correspond closely with the temperature maximizing performance and Darwinian fitness. Testing this co-adaptational hypothesis has been problematic for several reasons. First, reproductive fitness is the appropriate measure, but is difficult to measure in most animals. Second, no single fitness measure applies to all demographic situations, complicating interpretations. Here we test the co-adaptation hypothesis by studying an organism (Caenorhabditis elegans) in which both fitness and thermal preference can be reliably measured. RESULTS: We find that natural isolates of C. elegans display a range of mean thermal preferences and also vary in their thermal sensitivities for fitness. Hot-seeking isolates CB4854 and CB4857 prefer temperatures that favor population growth rate (r), whereas the cold-seeking isolate CB4856 prefers temperatures that favor Lifetime Reproductive Success (LRS). CONCLUSIONS: Correlations between fitness and thermal preference in natural isolates of C. elegans are driven primarily by isolate-specific differences in thermal preference. If these differences are the result of natural selection, then this suggests that the appropriate measure of fitness for use in evolutionary ecology studies might differ even within species, depending on the unique ecological and evolutionary history of each population. PMID- 21645396 TI - Telomerase expression and telomere length in breast cancer and their associations with adjuvant treatment and disease outcome. AB - INTRODUCTION: Telomere length plays important roles in maintaining genome stability and regulating cell replication and death. Telomerase has functions not only to extend telomere length but also to repair DNA damage. Studies have shown that telomerase may increase cancer cell resistance to DNA-damaging anticancer agents; tamoxifen may suppress telomerase expression in breast cancer cells. This study aimed to investigate the role of telomere length and telomerase activity in breast cancer prognosis. METHODS: qPCR and qRT-PCR were used to analyze telomere length and telomerase expression, respectively, in tumor samples of 348 breast cancer patients. Cox regression analysis was performed to examine telomere length and telomerase expression in association with disease-free survival and cause specific mortality. RESULTS: Telomere length had no relation to tumor features or disease outcomes. Telomerase expression was detected in 53% of tumors. Larger tumors or aggressive disease were more likely to have telomerase expression. Among patients treated with chemotherapy, high telomerase was found to be associated with increased risk of death (hazard ratio (HR) = 3.15; 95% CI: 1.34 to 7.40) and disease recurrence (HR = 2.04; 95% CI: 0.96 to 4.30) regardless of patient age, disease stage, tumor grade, histological type or hormone receptor status. Patients treated with endocrine therapy had different results regarding telomerase: high telomerase appeared to be associated with better survival outcomes. Telomerase expression made no survival difference in patients who received both chemotherapy and endocrine therapy. CONCLUSIONS: Overall, telomerase expression was not associated with disease outcome, but this finding may be masked by adjuvant treatment. Patients with high telomerase expression responded poorly to chemotherapy in terms of disease-free and overall survival, but fared better if treated with endocrine therapy. PMID- 21645397 TI - SIP1 is downregulated in hepatocellular carcinoma by promoter hypermethylation. AB - BACKGROUND: Smad interacting protein-1 is a transcription factor that is implicated in transforming growth factor-beta/bone morphogenetic protein signaling and a repressor of E-cadherin and human telomerase reverse transcriptase. It is also involved in epithelial-mesenchymal transition and tumorigenesis. However, genetic and epigenetic alterations of SIP1 have not been fully elucidated in cancers. In this study, we investigated mutations and promoter hypermethylation of the SIP1 gene in human hepatocellular carcinomas. METHODS: SIP1 expression was analyzed in HCC cell lines and primary tumors in comparison to normal and non-tumor liver tissues by using semi-quantitative RT PCR, quantitative real-time RT-PCR and immunohistochemistry. Mutation and deletion screening of the SIP1 gene were performed by direct sequencing in HCC derived cells. Restoration of SIP1 expression was sought by treating HCC cell lines with the DNA methyl transferase inhibitor, 5-AzaC, and the histone deacetylase inhibitor, TSA. SIP1 promoter methylation was analyzed by the combined bisulfite restriction analysis assay in in silico-predicted putative promoter and CpG island regions. RESULTS: We found that the expression of SIP1 was completely lost or reduced in five of 14 (36%) HCC cell lines and 17 of 23 (74%) primary HCC tumors. Immunohistochemical analysis confirmed that SIP1 mRNA downregulation was associated with decreased expression of the SIP1 protein in HCC tissues (82.8%). No somatic mutation was observed in SIP1 exons in any of the 14 HCC cell lines. Combined treatment with DNA methyl transferase and histone deacetylase inhibitors synergistically restored SIP1 expression in SIP1-negative cell lines. Analysis of three putative gene regulatory regions revealed tumor specific methylation in more than half of the HCC cases. CONCLUSIONS: Epigenetic mechanisms contribute significantly to the downregulation of SIP1 expression in HCC. This finding adds a new level of complexity to the role of SIP1 in hepatocarcinogenesis. PMID- 21645399 TI - Acute ST segment elevation during exercise stress echocardiography due to severe pulmonary hypertension. AB - A 51-year-old female undergoing an outpatient stress echocardiogram to evaluate atypical chest pain developed acute ST elevation in the anterior precordial leads on electrocardiogram following exercise. Echocardiography revealed a severe rise in pulmonary artery systolic pressure (PASP) with marked right ventricular (RV) enlargement and interventricular septum flattening. Subsequently, cardiac catherization confirmed an exercise-induced elevation in PASP and diagnosed pulmonary arterial hypertension without evidence of coronary artery disease. This case suggests that an acute elevation in pulmonary artery pressure with RV dilation may be a potential cause of acute ST elevation during stress testing. PMID- 21645398 TI - Rotavirus NSP4: Cell type-dependent transport kinetics to the exofacial plasma membrane and release from intact infected cells. AB - BACKGROUND: Rotavirus NSP4 localizes to multiple intracellular sites and is multifunctional, contributing to RV morphogenesis, replication and pathogenesis. One function of NSP4 is the induction of early secretory diarrhea by binding surface receptors to initiate signaling events. The aims of this study were to determine the transport kinetics of NSP4 to the exofacial plasma membrane (PM), the subsequent release from intact infected cells, and rebinding to naive and/or neighboring cells in two cell types. METHODS: Transport kinetics was evaluated using surface-specific biotinylation/streptavidin pull-downs and exofacial exposure of NSP4 was confirmed by antibody binding to intact cells, and fluorescent resonant energy transfer. Transfected cells similarly were monitored to discern NSP4 movement in the absence of infection or other viral proteins. Endoglycosidase H digestions, preparation of CY3- or CY5- labeled F(ab)2 fragments, confocal imaging, and determination of preferential polarized transport employed standard laboratory techniques. Mock-infected, mock biotinylated and non-specific antibodies served as controls. RESULTS: Only full length (FL), endoglycosidase-sensitive NSP4 was detected on the exofacial surface of two cell types, whereas the corresponding cell lysates showed multiple glycosylated forms. The C-terminus of FL NSP4 was detected on exofacial-membrane surfaces at different times in different cell types prior to its release into culture media. Transport to the PM was rapid and distinct yet FL NSP4 was secreted from both cell types at a time similar to the release of virus. NSP4 containing, clarified media from both cells bound surface molecules of naive cells, and imaging showed secreted NSP4 from one or more infected cells bound neighboring cell membranes in culture. Preferential sorting to apical or basolateral membranes also was distinct in different polarized cells. CONCLUSIONS: The intracellular transport of NSP4 to the PM, translocation across the PM, exposure of the C-terminus on the cell surface and subsequent secretion occurs via an unusual, complex and likely cell-dependent process. The exofacial exposure of the C-terminus poses several questions and suggests an atypical mechanism by which NSP4 traverses the PM and interacts with membrane lipids. Mechanistic details of the unconventional trafficking of NSP4, interactions with host-cell specific molecules and subsequent release require additional study. PMID- 21645400 TI - Efficient unfolding pattern recognition in single molecule force spectroscopy data. AB - BACKGROUND: Single-molecule force spectroscopy (SMFS) is a technique that measures the force necessary to unfold a protein. SMFS experiments generate Force Distance (F-D) curves. A statistical analysis of a set of F-D curves reveals different unfolding pathways. Information on protein structure, conformation, functional states, and inter- and intra-molecular interactions can be derived. RESULTS: In the present work, we propose a pattern recognition algorithm and apply our algorithm to datasets from SMFS experiments on the membrane protein bacterioRhodopsin (bR). We discuss the unfolding pathways found in bR, which are characterised by main peaks and side peaks. A main peak is the result of the pairwise unfolding of the transmembrane helices. In contrast, a side peak is an unfolding event in the alpha-helix or other secondary structural element. The algorithm is capable of detecting side peaks along with main peaks.Therefore, we can detect the individual unfolding pathway as the sequence of events labeled with their occurrences and co-occurrences special to bR's unfolding pathway. We find that side peaks do not co-occur with one another in curves as frequently as main peaks do, which may imply a synergistic effect occurring between helices. While main peaks co-occur as pairs in at least 50% of curves, the side peaks co occur with one another in less than 10% of curves. Moreover, the algorithm runtime scales well as the dataset size increases. CONCLUSIONS: Our algorithm satisfies the requirements of an automated methodology that combines high accuracy with efficiency in analyzing SMFS datasets. The algorithm tackles the force spectroscopy analysis bottleneck leading to more consistent and reproducible results. PMID- 21645401 TI - Determining research knowledge infrastructure for healthcare systems: a qualitative study. AB - BACKGROUND: This study examines research knowledge infrastructures (RKIs) found in health systems. An RKI is defined as any instrument (i.e., programs, interventions, tools) implemented in order to facilitate access, dissemination, exchange, and/or use of evidence in healthcare organisations. Based on an environmental scan (17 key informant interviews) and scoping review (26 studies), we found support for a framework that we developed that outlines components that a health system can have in its RKI. The broad domains are climate for research use, research production, activities used to link research to action, and evaluation.The objective of the current study is to profile the RKI of three types of health system organisations--regional health authorities, primary care practices, and hospitals--in two Canadian provinces to determine the current mix of components these organisations have in their RKI, their experience with these components, and their views about future RKI initiatives. METHODS: This study will include semistructured telephone interviews with a purposive sample region of a senior management team member, library/resource centre manager, and a 'knowledge broker' in three regional health authorities, five or six purposively sampled hospitals, and five or six primary care practices in Ontario and Quebec, for a maximum of 71 interviewees. The interviews will explore (a) which RKI components have proven helpful, (b) barriers and facilitators in implementing RKI components, and (c) views about next steps in further development of RKIs. DISCUSSION: This is the first qualitative examination of potential RKI efforts that can increase the use of research evidence in health system decision making. We anticipate being able to identify broadly applicable insights about important next steps in building effective RKIs. Some of the identified RKI components may increase the use of research evidence by decision makers, which may then lead to more informed decisions. PMID- 21645402 TI - Patterns of anti-malarial drug treatment among pregnant women in Uganda. AB - BACKGROUND: Prompt use of an effective anti-malarial drug is essential for controlling malaria and its adverse effects in pregnancy. The World Health Organization recommends an artemisinin-based combination therapy as the first line treatment of uncomplicated malaria in the second and third trimesters of pregnancy. The study objective was to determine the degree to which presumed episodes of uncomplicated symptomatic malaria in pregnancy were treated with a recommended anti-malarial regimen in a region of Uganda. METHODS: Utilizing a population-based random sample, we interviewed women living in Jinja, Uganda who had been pregnant in the past year. RESULTS: Self-reported malaria during the index pregnancy was reported among 67% (n = 334) of the 500 participants. Among the 637 self-reported episodes of malaria, an anti-malarial drug was used for treatment in 85% of the episodes. Use of a currently recommended treatment in the first trimester was uncommon (5.6%). A contraindicated anti-malarial drug (sulphadoxine-pyrimethamine and/or artemether-lumefantrine) was involved in 70% of first trimester episodes. Recommended anti-malarials were used according to the guidelines in only 30.1% of all second and third trimester episodes. CONCLUSIONS: Self-reported malaria was extremely common in this population and adherence to treatment guidelines for the management of malaria in pregnancy was poor. Use of artemether-lumefantrine combined with non-recommended anti-malarials was common practice. Overuse of anti-malarial drugs, especially ones that are no longer recommended, undermines malaria control efforts by fueling the spread of drug resistance and delaying appropriate treatment of non-malarial febrile illnesses. Improved diagnostic capacity is essential to ultimately improving the management of malaria-like symptoms during pregnancy and appropriate use of currently available anti-malarials. PMID- 21645403 TI - Early risk factors for being a bully, victim, or bully/victim in late elementary and early secondary education. The longitudinal TRAILS study. AB - BACKGROUND: Data regarding the impact of early risk factors on later involvement in bullying are scarce. We investigated the impact of preschool behaviors, family characteristics (socio-economic status, family breakup) and parental mental health on bullying and victimization at age 11 (T1) and age 13.5 (T2). METHODS: longitudinal data from a subsample of the TRacking Adolescents' Individual Lives Survey (TRAILS) (T1: N = 982; T2: N = 977). TRAILS is a prospective study of adolescent mental health in a mixed urban and rural region of the Netherlands. At T1 parents reported on family characteristics, parental mental health and retrospectively on children's preschool behavior at age 4-5. Schoolmates reported involvement of adolescents in bullying or victimization at T1 and T2. RESULTS: Children with preschool anxiety were less likely to be bully/victim at T1. Children with preschool aggressiveness were more likely to be bully (T1), bully/victim (T1 and T2) and victim (T2) and children with good preschool motor functioning were more likely to be bully (T1) and less likely to be victim (T1 and T2). Children from low socioeconomic status families were more likely be to be bully, victim, or bully/victim and less likely to be uninvolved both at T1 and T2. Finally, children from intact two parent families were more likely to be uninvolved at T2. CONCLUSION: Preschool behavioral, emotional and motor problems, socioeconomic status, and family breakup are related to involvement in bullying at a later age. Prevention of bullying and its consequences can be enhanced by focusing on risk groups in early life. PMID- 21645404 TI - In vivo UVA irradiation of mouse is more efficient in promoting pulmonary melanoma metastasis than in vitro. AB - BACKGROUND: We have previously shown in vitro that UVA increases the adhesiveness of mouse B16-F1 melanoma cells to endothelium.We have also shown in vivo that UVA exposure of C57BL/6 mice, i.v. injected with B16-F1 cells, increases formation of pulmonary colonies of melanoma. The aim of the present animal study was to confirm the previously observed in vivo UVA effect and to determine whether in vitro UVA-exposure of melanoma cells, prior the i.v. injection, will have an enhancing effect on the pulmonary colonization capacity of melanoma cells. As a second aim, UVA-derived immunosuppression was determined. METHODS: Mice were i.v. injected with B16-F1 cells into the tail vein and then immediately exposed to UVA. Alternatively, to study the effect of UVA-induced adhesiveness on the colonization capacity of B16-F1 melanoma, cells were in vitro exposed prior to i.v. injection. Fourteen days after injection, lungs were collected and the number of pulmonary nodules was determined under dissecting microscope. The UVA derived immunosuppression was measured by standard contact hypersensitivity assay. RESULTS AND DISCUSSION: Obtained results have confirmed that mice, i.v. injected with B16-F1 cells and thereafter exposed to UVA, developed 4-times more of melanoma colonies in lungs as compared with the UVA non-exposed group (p < 0.01). The in vitro exposure of melanoma cells prior to their injection into mice, led only to induction of 1.5-times more of pulmonary tumor nodules, being however a statistically non-significant change. The obtained results postulate that the UVA-induced changes in the adhesive properties of melanoma cells do not alone account for the 4-fold increase in the pulmonary tumor formation. Instead, it suggests that some systemic effect in a mouse might be responsible for the increased metastasis formation. Indeed, UVA was found to induce moderate systemic immunosuppression, which effect might contribute to the UVA-induced melanoma metastasis in mice lungs. PMID- 21645405 TI - Increased expression of Ero1L-alpha in healing fetal wounds. AB - BACKGROUND: Adult mammalian tissues heal injury to the skin with formation of scar; this process quickly seals an injured area, however, excessive scar formation can become a source of persistent pathology, interfering with multiple vital functions. In contrast, mammalian fetal tissue can heal without scar formation. We previously sought to model scarless healing in a rabbit fetal skin wound and identified gene products differentially expressed during fetal wound healing through PCR suppression subtractive hybridization (PCR SSH). One of these transcripts, previously identified simply as clone 11, showed putative increased expression in wounded fetal skin. This study establishes its identity as Ero1L alpha and confirms its elevated expression in healing fetal wounds. FINDINGS: After obtaining further sequence by 5' rapid amplification of cloned ends (RACE) we find that clone 11 is Ero1L-alpha. We determined that clone 11, a differentially expressed transcript in fetal wound healing, comprises the 3' untranslated region (UTR) of an approximately 4 kb transcript in rabbit tissues that corresponds to Ero1L-alpha. We showed that Ero1L-alpha is expressed predominantly as two transcripts in rabbit skin, namely a 1.6 kb transcript and the 4.0 kb transcript recovered in our PCR SSH screen via its 3' UTR sequence. However, a third transcript of 2.9 kb was also detected in Northern blots and was subsequently cloned and confirmed by 3' RACE. Knockdown of the clone 11 sequence in rabbit adult fibroblasts via siRNA resulted in significantly decreased Ero1L alpha message expression. Increased expression of clone 11 (Ero1L-alpha) in a variety of cell types during the wound healing response was also confirmed by in situ hybridization. CONCLUSIONS: Ero1L-alpha is one of the previously unknown clones identified in a PCR SSH screen for genes differentially expressed in fetal wounded tissue. In situ hybridization confirms that Ero1L-alpha shows increased expression in multiple cell types after wounding of the fetal integument. PMID- 21645406 TI - Error-dependent modulation of speech-induced auditory suppression for pitch shifted voice feedback. AB - BACKGROUND: The motor-driven predictions about expected sensory feedback (efference copies) have been proposed to play an important role in recognition of sensory consequences of self-produced motor actions. In the auditory system, this effect was suggested to result in suppression of sensory neural responses to self produced voices that are predicted by the efference copies during vocal production in comparison with passive listening to the playback of the identical self-vocalizations. In the present study, event-related potentials (ERPs) were recorded in response to upward pitch shift stimuli (PSS) with five different magnitudes (0, +50, +100, +200 and +400 cents) at voice onset during active vocal production and passive listening to the playback. RESULTS: Results indicated that the suppression of the N1 component during vocal production was largest for unaltered voice feedback (PSS: 0 cents), became smaller as the magnitude of PSS increased to 200 cents, and was almost completely eliminated in response to 400 cents stimuli. CONCLUSIONS: Findings of the present study suggest that the brain utilizes the motor predictions (efference copies) to determine the source of incoming stimuli and maximally suppresses the auditory responses to unaltered feedback of self-vocalizations. The reduction of suppression for 50, 100 and 200 cents and its elimination for 400 cents pitch-shifted voice auditory feedback support the idea that motor-driven suppression of voice feedback leads to distinctly different sensory neural processing of self vs. non-self vocalizations. This characteristic may enable the audio-vocal system to more effectively detect and correct for unexpected errors in the feedback of self produced voice pitch compared with externally-generated sounds. PMID- 21645407 TI - Adolescents' leisure activities, parental monitoring and cigarette smoking--a cross-sectional study. AB - BACKGROUND: Adolescent participation in leisure activities is developmentally beneficial, but certain activities may increase health compromising behaviours, such as tobacco smoking. A limited range of leisure activities has been studied, with little research on out-of-school settings where parental supervision is a potential protective factor. Tobacco smoking is an important, potentially modifiable health determinant, so understanding associations between adolescent leisure activities, parental monitoring, demographic factors and daily smoking may inform preventive strategies. These associations are reported for a New Zealand adolescent sample. METHODS: Randomly selected schools (n = 145) participated in the 2006 Youth In-depth Survey, a national, biennial study of Year 10 students (predominantly 14-15 years). School classes were randomly selected and students completed a self-report questionnaire in class time. Adjustment for clustering at the school level was included in all analyses. Since parental monitoring and demographic variables potentially confound relations between adolescent leisure activities and smoking, variables were screened before multivariable modelling. Given prior indications of demographic differences, gender and ethnic specific regression models were built. RESULTS AND DISCUSSION: Overall, 8.5% of the 3,161 students were daily smokers, including more females (10.5%) than males (6.5%). In gender and ethnic specific multivariate analysis of associations with daily smoking (adjusted for age, school socioeconomic decile rating, leisure activities and ethnicity or gender, respectively), parental monitoring exhibited a consistently protective, dose response effect, although less strongly among Maori. Attending a place of worship and going to the movies were protective for non-Maori, as was watching sports, whereas playing team sport was protective for all, except males. Attending a skate park was a risk factor for females and Maori which demonstrated a strong dose response effect. CONCLUSIONS: There were significant differences in the risk of daily smoking across leisure activities by gender and ethnicity. This reinforces the need to be alert for, and respond to, gender and ethnic differences in the pattern of risk and protective factors. However, given the consistently protective, dose response effect of parental monitoring, our findings confirm that assisting oversight of adolescent leisure activities may be a key component in public health policy and prevention programmes. PMID- 21645408 TI - Aintegumenta and Aintegumenta-Like6 regulate auxin-mediated flower development in Arabidopsis. AB - BACKGROUND: Two related genes encoding AP2/ERF-type transcription factors, AINTEGUMENTA (ANT) and AINTEGUMENTA-LIKE6 (AIL6), are important regulators of floral growth and patterning in Arabidopsis. Evidence suggests that these genes promote several aspects of flower development in response to auxin. To investigate the interplay of ANT, AIL6 and auxin during floral development, I have examined the phenotypic consequences of disrupting polar auxin transport in ant, ail6 and ant ail6 mutants by either genetic or chemical means. RESULTS: Plants containing mutations in ANT or AIL6 alone or in both genes together exhibit increased sensitivity to disruptions in polar auxin transport. Both genes promote shoot growth, floral meristem initiation and floral meristem patterning in combination with auxin transport. However, differences in the responses of ant and ail6 single mutants to perturbations in auxin transport suggest that these two genes also have non-overlapping activities in each of these developmental processes. CONCLUSIONS: The enhanced sensitivity of ant and ail6 mutants to alterations in polar auxin transport suggests that these mutants have defects in some aspect of auxin physiology. The inability of ant ail6 double mutants to initiate flowers in backgrounds disrupted for auxin transport confirm the proposed roles for these two genes in floral meristem initiation. PMID- 21645409 TI - X-linked inhibitor of apoptosis positive nuclear labeling: a new independent prognostic biomarker of breast invasive ductal carcinoma. AB - BACKGROUND: It's well recognized that X-linked inhibitor of apoptosis (XIAP) was the most potent caspase inhibitor and second mitochondria-derived activator of caspase (Smac) was the antagonist of XIAP. Experiments in vitro identified that down regulation of XIAP expression or applying Smac mimics could sensitize breast cancer cells to chemotherapeutics and promote apoptosis. However, expression status and biologic or prognostic significance of XIAP/Smac in breast invasive ductal carcinoma (IDC) were not clear. The present study aimed to investigate relationship among expression status of XIAP/Smac, apoptosis index (AI), clinicopathologic parameters and prognosis in IDC. METHODS: Immunohistochemistry and TUNEL experiment were performed to detect expression of XIAP, Smac, ER, PR, HER2 and AI in 102 cases of paraffin-embedded IDC samples respectively. Expression of XIAP/Smac were also detected in limited 8 cases of fresh IDC specimens with Western blot. RESULTS: Positive ratio and immunoscore of XIAP was markedly higher than Smac in IDC (P<0.0001). It was noteworthy that 44 cases of IDC were positive in nuclear for XIAP, but none was for Smac. Expression status of Smac was more prevalent in HER2 positive group than negative group (P<0.0001) and AI was positively correlated with HER2 protein expression (rs=0.265, P=0.017). The present study first revealed that XIAP positive nuclear labeling (XIAP-N), but not cytoplasmic staining (XIAP-C), was the apoptotic marker correlated significantly with patients' shortened overall survival (P=0.039). Survival analysis demonstrated that XIAP-N was a new independent prognostic factor except for patient age and lymph node status. CONCLUSION: Disturbed balance of expression between XIAP and Smac probably contributed to carcinogenesis and XIAP positive nuclear labeling was a new independent prognostic biomarker of breast IDC. PMID- 21645410 TI - A comparison of two headless compression screws for operative treatment of scaphoid fractures. AB - PURPOSE: The purpose of this study was to compare the interfragmentary compression force across a simulated scaphoid fracture by two commonly used compression screw systems; the Acutrak 2 Standard and the 3.0 mm Synthes headless compression screw. METHODS: Sixteen (8 pairs; 6 female, 2 male) cadaver scaphoids were randomly assigned to receive either the Acutrak 2 or Synthes screw with the contralateral scaphoid designated to receive the opposite. Guide wires were inserted under fluoroscopic control. Following transverse osteotomy, the distal and proximal fragments were placed on either side of a custom load cell, to measure interfragmentary compression. Screws were placed under fluoroscopic control using the manufacturer's recommended surgical technique. Compressive forces were measured during screw insertion. Recording continued for an additional 60s in order to measure any loss of compression after installation was complete. The peak and final interfragmentary compression were recorded and paired t-tests performed. RESULTS: The mean peak compression generated by the Acutrak 2 Standard was greater than that produced by the Synthes compression screw (103.9 +/- 33.2 N vs. 88.7 +/- 38.6 N respectively, p = 0.13). The mean final interfragmentary compression generated by the Acutrak 2 screw (68.6 +/- 36.4 N) was significantly greater (p = 0.04) than the Synthes screw (37.2 +/- 26.8 N). Specimens typically reached a steady state of compression by 120-150s after final tightening. CONCLUSION: Peak interfragmentary compression observed during screw installation was similar for both screw systems. However, the mean interfragmentary compression generated by the Acutrak 2 Standard was significantly greater. Our study demonstrates that the Synthes headless compression screw experienced a greater loss of interfragmentary compressive force from the time of installation to the final steady state compression level. The higher post-installation compression of the Acutrak 2 Standard may be attributable to the greater number of threads throughout the entire length of the screw. The clinical significance of these results, are, at this point uncertain. We do demonstrate that a fully threaded design offers a more reliable compression that may translate to more predictable bony union. PMID- 21645411 TI - Molecular profile and copy number analysis of sporadic colorectal cancer in Taiwan. AB - BACKGROUND: Colorectal cancer (CRC) is a major health concern worldwide, and recently becomes the most common cancer in Asia. The case collection of this study is one of the largest sets of CRC in Asia, and serves as representative data for investigating genomic differences between ethnic populations. We took comprehensive and high-resolution approaches to compare the clinicopathologic and genomic profiles of microsatellite instability (MSI) vs. microsatellite stability (MSS) in Taiwanese sporadic CRCs. METHODS: 1,173 CRC tumors were collected from the Taiwan population, and sequencing-based microsatellite typing assay was used to determine MSI and MSS. Genome-wide SNP array was used to detect CN alterations in 16 MSI-H and 13 MSS CRCs and CN variations in 424 general controls. Gene expression array was used to evaluate the effects of CN alterations, and quantitative PCR methods were used to replicate the findings in independent clinical samples. RESULTS: These 1,173 CRC tumors can be classified into 75 high frequency MSI (MSI-H) (6.4%), 96 low-frequency MSI (8.2%) and 1,002 MSS (85.4%). Of the 75 MSI-H tumors, 22 had a BRAF mutation and 51 showed MLH1 promoter hypermethylation. There were distinctive differences in the extent of CN alterations between CRC MSS and MSI-H subtypes (300 Mb vs. 42 Mb per genome, p value < 0.001). Also, chr7, 8q, 13 and 20 gains, and 8p and 18 losses were frequently found in MSS but not in MSI-H. Nearly a quarter of CN alterations were smaller than 100 kb, which might have been missed in previous studies due to low resolution technology. 514 expressed genes showed CN differences between subtypes, and 271 of them (52%) were differentially expressed. CONCLUSIONS: Sporadic CRCs with MSI-H displayed distinguishable clinicopathologic features, which differ from those of MSS. Genomic profiling of the two types of sporadic CRCs revealed significant differences in the extent and distribution of CN alterations in the cancer genome. More than half of expressed genes showing CN differences can directly contribute to their expressional diversities, and the biological functions of the genes associated with CN changes in sporadic CRCs warrant further investigation to establish their possible clinical implications. PMID- 21645412 TI - Sphingosine-1-phosphate promotes the differentiation of human umbilical cord mesenchymal stem cells into cardiomyocytes under the designated culturing conditions. AB - BACKGROUND: It is of growing interest to develop novel approaches to initiate differentiation of mesenchymal stem cells (MSCs) into cardiomyocytes. The purpose of this investigation was to determine if Sphingosine-1-phosphate (S1P), a native circulating bioactive lipid metabolite, plays a role in differentiation of human umbilical cord mesenchymal stem cells (HUMSCs) into cardiomyocytes. We also developed an engineered cell sheet from these HUMSCs derived cardiomyocytes by using a temperature-responsive polymer, poly(N-isopropylacrylamide) (PIPAAm) cell sheet technology. METHODS: Cardiomyogenic differentiation of HUMSCs was performed by culturing these cells with either designated cardiomyocytes conditioned medium (CMCM) alone, or with 1 MUM S1P; or DMEM with 10% FBS + 1 MUM S1P. Cardiomyogenic differentiation was determined by immunocytochemical analysis of expression of cardiomyocyte markers and patch clamping recording of the action potential. RESULTS: A cardiomyocyte-like morphology and the expression of alpha-actinin and myosin heavy chain (MHC) proteins can be observed in both CMCM culturing or CMCM+S1P culturing groups after 5 days' culturing, however, only the cells in CMCM+S1P culture condition present cardiomyocyte-like action potential and voltage gated currents. A new approach was used to form PIPAAm based temperature responsive culture surfaces and this successfully produced cell sheets from HUMSCs derived cardiomyocytes. CONCLUSIONS: This study for the first time demonstrates that S1P potentiates differentiation of HUMSCs towards functional cardiomyocytes under the designated culture conditions. Our engineered cell sheets may provide a potential for clinically applicable myocardial tissues should promote cardiac tissue engineering research. PMID- 21645413 TI - Proteomics research on muscle-invasive bladder transitional cell carcinoma. AB - BACKGROUND: Aimed to facilitate candidate biomarkers selection and improve network-based multi-target therapy, we perform comparative proteomics research on muscle-invasive bladder transitional cell carcinoma. Laser capture microdissection was used to harvest purified muscle-invasive bladder cancer cells and normal urothelial cells from 4 paired samples. Two-dimensional liquid chromatography tandem mass spectrometry was used to identify the proteome expression profile. The differential proteins were further analyzed using bioinformatics tools and compared with the published literature. RESULTS: A total of 885/890 proteins commonly appeared in 4 paired samples. 295/337 of the 488/493 proteins that specific expressed in tumor/normal cells own gene ontology (GO) cellular component annotation. Compared with the entire list of the international protein index (IPI), there are 42/45 GO terms exhibited as enriched and 9/5 exhibited as depleted, respectively. Several pathways exhibit significantly changes between cancer and normal cells, mainly including spliceosome, endocytosis, oxidative phosphorylation, etc. Finally, descriptive statistics show that the PI Distribution of candidate biomarkers have certain regularity. CONCLUSIONS: The present study identified the proteome expression profile of muscle-invasive bladder cancer cells and normal urothelial cells, providing information for subcellular pattern research of cancer and offer candidate proteins for biomarker panel and network-based multi-target therapy. PMID- 21645414 TI - Characterization of killer immunoglobulin-like receptor genetics and comprehensive genotyping by pyrosequencing in rhesus macaques. AB - BACKGROUND: Human killer immunoglobulin-like receptors (KIRs) play a critical role in governing the immune response to neoplastic and infectious disease. Rhesus macaques serve as important animal models for many human diseases in which KIRs are implicated; however, the study of KIR activity in this model is hindered by incomplete characterization of KIR genetics. RESULTS: Here we present a characterization of KIR genetics in rhesus macaques (Macaca mulatta). We conducted a survey of KIRs in this species, identifying 47 novel full-length KIR sequences. Using this expanded sequence library to build upon previous work, we present evidence supporting the existence of 22 Mamu-KIR genes, providing a framework within which to describe macaque KIRs. We also developed a novel pyrosequencing-based technique for KIR genotyping. This method provides both comprehensive KIR genotype and frequency estimates of transcript level, with implications for the study of KIRs in all species. CONCLUSIONS: The results of this study significantly improve our understanding of macaque KIR genetic organization and diversity, with implications for the study of many human diseases that use macaques as a model. The ability to obtain comprehensive KIR genotypes is of basic importance for the study of KIRs, and can easily be adapted to other species. Together these findings both advance the field of macaque KIRs and facilitate future research into the role of KIRs in human disease. PMID- 21645415 TI - The analgesic effect of electroacupuncture on acute thermal pain perception--a central neural correlate study with fMRI. AB - BACKGROUND: Electrical acupuncture (EA) has been utilized in acute pain management. However, the neuronal mechanisms that lead to the analgesic effect are still not well defined. The current study assessed the intensity [optimal EA (OI-EA) vs. minimal EA (MI-EA)] effect of non-noxious EA on supraspinal regions related to noxious heat pain (HP) stimulation utilizing an EA treatment protocol for acute pain and functional magnetic resonance imaging (fMRI) with correlation in behavioral changes. Subjects underwent five fMRI scanning paradigms: one with heat pain (HP), two with OI-EA and MI-EA, and two with OI-EA and HP, and MI-EA and HP. RESULTS: While HP resulted in activations (excitatory effect) in supraspinal areas known for pain processing and perception, EA paradigms primarily resulted in deactivations (suppressive effect) in most of these corresponding areas. In addition, OI-EA resulted in a more robust supraspinal sedative effect in comparison to MI-EA. As a result, OI-EA is more effective than MI-EA in suppressing the excitatory effect of HP in supraspinal areas related to both pain processing and perception. CONCLUSION: Intensities of EA plays an important role in modulating central pain perception. PMID- 21645416 TI - Expression of miR-1, miR-133a, miR-133b and miR-206 increases during development of human skeletal muscle. AB - BACKGROUND: MicroRNAs (miRNAs) are small RNA molecules that post transcriptionally regulate gene expression and have been shown to play an important role during development. miR-1, miR-133a, miR-133b and miR-206 are expressed in muscle tissue and induced during muscle cell differentiation, a process that directs myoblasts to differentiate into mature myotubes, which are organized into myofibers. Although miR-1, miR-133a, miR-133b and miR-206 are well studied in muscle, there is no information about their expression and function during human development. The purpose of this study was to determine the profile of these miRNAs in muscle cells isolated from different stages of human development. RESULTS: We examined the levels of miR-1, miR-133a, miR-133b and miR 206 during the development of human foetus. All four miRNA levels were found increased during late stages of human foetal muscle development. Increases in the expression levels of these miRNAs were proportional to the capacity of myoblasts to form myotubes. Changes in miRNA levels during human foetal development were accompanied by endogenous alterations in their known targets and also in their inducer, MyoD. Ectopic MyoD expression caused an induction of muscle cell differentiation in vitro, accompanied by an increase in the levels of miR-1, miR 133a, miR-133b and miR-206. CONCLUSIONS: This study provides data about the profile of four miRNAs in human muscle cells isolated during different stages of foetal development. These results may shed light on the differentiation of muscle cells and regulation of muscle formation through miRNAs, during the development of human foetus. PMID- 21645417 TI - Clinical symptoms and chemotherapy completion in elderly patients with newly diagnosed acute leukemia: a retrospective comparison study with a younger cohort. AB - BACKGROUND: Cancer affects older adults disproportionately. The disease is often difficult to diagnose and treat due to co-morbidities and performance status, and patients tend to discontinue chemotherapy prematurely. There are no systemic studies of the reasons and factors that create a higher withdrawal rate in older acute leukemia patients. This study tried to understand the initial characteristics, blood counts and bone marrow measurements in older acute leukemia patients by comparing them with a younger group to provide information and assistance in early clinical diagnosis, treatment and reasons for treatment withdrawal. METHODS: Using retrospective medical record reviews, we examined clinical characteristics and chemotherapy completion status in the patients of two groups (age >= 60, n = 183 and age <60, n = 183) who were diagnosed with acute leukemia for the first time and were hospitalized in Union Hospital Affiliated with Fujian Medical University from 2004 to 2008. RESULTS: There were no statistical differences in initial presenting symptoms of fatigue (67.2% vs. 57.9%, P>0.05) and pallor (53% vs. 59.6%, P>0.05) between the two groups, but older patients demonstrated more underlying diseases including lung infections (25.7%, P = <0.001), cardiovascular disease (4.4%, P = 0.007), and hypertension (20.8%, P =< 0.001). The complete remission rate after chemotherapy (1 to 2 courses) was 49.5% in the older group and 66.7% in the younger group (chi2 = 6.202, P = 0.013). The percentage of patients age 60 and older who prematurely discontinued chemotherapy (50.3%), mainly due to the influences of traditional Chinese concept of critical illness, financial difficulties, and intolerance to adverse reactions to chemotherapy, was significantly higher than that of younger patients (37.7%) (chi2 = 5.866, P = 0.015). CONCLUSIONS: A comprehensive approach to diagnosis, treatment selection, and toxicity management, and implementing strategies to enhance treatment compliance may improve outcomes in older adults with acute leukemia. PMID- 21645418 TI - Method for Assigning Priority Levels in Acute Care (MAPLe-AC) predicts outcomes of acute hospital care of older persons--a cross-national validation. AB - BACKGROUND: Although numerous risk factors for adverse outcomes for older persons after an acute hospital stay have been : identified, a decision making tool combining all available information in a clinically meaningful way would be helpful for daily hospital practice. The purpose of this study was to evaluate the ability of the Method for Assigning Priority Levels for Acute Care (MAPLe-AC) to predict adverse outcomes in acute care for older people and to assess its usability as a decision making tool for discharge planning. METHODS: Data from a prospective multicenter study in five Nordic acute care hospitals with information from admission to a one year follow-up of older acute care patients were compared with a prospective study of acute care patients from admission to discharge in eight hospitals in Canada. The interRAI Acute Care assessment instrument (v1.1) was used for data collection. Data were collected during the first 24 hours in hospital, including pre-morbid and admission information, and at day 7 or at discharge, whichever came first. Based on this information a crosswalk was developed from the original MAPLe algorithm for home care settings to acute care (MAPLe-AC). The sample included persons 75 years or older who were admitted to acute internal medical services in one hospital in each of the five Nordic countries (n = 763) or to acute hospital care either internal medical or combined medical-surgical services in eight hospitals in Ontario, Canada (n = 393). The outcome measures considered were discharge to home, discharge to institution or death. Outcomes in a 1-year follow-up in the Nordic hospitals were: living at home, living in an institution or death, and survival. Logistic regression with ROC curves and Cox regression analyses were used in the analyses. RESULTS: Low and mild priority levels of MAPLe-AC predicted discharge home and high and very high priority levels predicted adverse outcome at discharge both in the Nordic and Canadian data sets, and one-year outcomes in the Nordic data set. The predictive accuracy (AUC's) of MAPLe-AC's was higher for discharge outcome than one year outcome, and for discharge home in Canadian hospitals but for adverse outcome in Nordic hospitals. High and very high priority levels in MAPLe AC were also predictive of days to death adjusted for diagnoses in survival models. CONCLUSION: MAPLe-AC is a valid algorithm based on risk factors that predict outcomes of acute hospital care. It could be a helpful tool for early discharge planning although further testing for active use in clinical practice is still needed. PMID- 21645419 TI - Willingness to pay for municipality hospital services in rural Japan: a contingent valuation study. AB - BACKGROUND: The Japanese healthcare system has undergone reforms to address the struggles that municipality hospitals face. Reform guidelines clearly define criteria for administrative improvement. However, criteria to evaluate the demand for healthcare provisions in rural Japan, including the needs of rural residents for municipality hospitals in particular have not been specified. The purpose of this paper is to measure residents' willingness to pay (WTP) for municipality hospital services using the contingent valuation method, and to evaluate municipality hospital valuation on the basis of WTP. K town, located in the Hokkaido prefecture of Japan, was selected as the location for this study. Participants were recruited by a town hall healthcare administrator, hospital and clinic staff, and a local dentist. Participants were asked what amount they would be willing to pay as taxes to continue accessing the services of the municipality hospital for one year by using open-ended questions in face-to-face interviews. FINDINGS: Forty-eight residents were initially recruited, and 40 participants were selected for the study (response rate 83%). As compared to K town's population, this data slanted toward the elderly, although there was no significant difference in frequency among the characteristics. The median WTP was estimated at 39,484 yen ($438.71), with a 95% confidence interval 27,806-55,437 yen ($308.95-615.96). Logistic regression revealed no significant factors affecting WTP. CONCLUSIONS: If the total amount of residents' WTP for the municipality hospital were to be estimated by this result, it would calculate with 129,586,000 yen ($1,439,844). This is approximately equal to the amount of money to be transferred from the general account of the government of K town, more than one-half of the town tax of K town, and about two-fold in comparison to Japan as a whole. This showed that K town's residents placed a high valuation on the municipality hospital, which nearly equalled the amount that the K town government provided to the municipality hospital to cover its annual deficit. K town residents had come to expect not only general clinical practice, but also emergency medical services and night practice provided by their own town's municipality hospital. WTP can be used as a measure of hospital evaluation because it reflects the importance of the hospital to the residents in its region. PMID- 21645420 TI - Characteristics and PD-1 expression of peripheral CD4+CD127loCD25hiFoxP3+ Treg cells in chronic HCV infected-patients. AB - BACKGROUND: Both regulatory T cells (Tregs) and PD-1/PD-L1 pathway were critically involved in HCV viral persistence. However, the association between them was not well investigated. Herein, we aimed to investigate the distributional profiles of Tregs subsets and association between PD-1 expression on these subsets and development of HCV long-term persistence. METHODS: CD45RA and CD27 were employed to separate peripheral Tregs as naive/central memory/effector memory/effector subsets. The phenotypic characteristics and PD-1 expression of Tregs were studied by flow cytometry. RESULTS: In the present study, the majority of Tregs was identified as central memory phenotype in chronic hepatitis C patients compared with nearly equal contribution of naive and central memory subsets in healthy individuals. PD-1 expression was elevated in all CD4+ T cell subset in chronic HCV infected patients, including Tregs. Of note, higher level of PD-1 expression was found on TEM- and effector-Treg than naive- and TCM-Tregs subsets. The ratio of TEM-Tregs/naive-Tregs and TEM Tregs/TCM-Tregs regarding to PD-1 MFI were significantly lower in CHC patients compared to controls. CONCLUSIONS: Our study indicated that distinctive characteristics of PD-1 expression on Tregs in HCV infection suggests associated with impaired adaptive immunity as well as viral long-term persistence. The cross talk between Treg cells and PD-1 induced inhibition in chronic HCV infection deserved further exploration for HCV infection associated immune pathogenesis. PMID- 21645421 TI - Molecular epidemiology of novel swine origin influenza virus (S-OIV) from Gwalior, India, 2009. AB - BACKGROUND: The H1N1pandemic virus is a newly emergent human influenza A virus that is closely related to a number of currently circulating pig viruses in the 'classic North American' and 'Eurasian' swine influenza virus lineages and thus referred as S-OIV. Since the first reports of the virus in humans in April 2009, H1N1 virus has spread to 168 countries and overseas territories. India also witnessed severe H1N1 pandemic virus epidemic with considerable morbidity and mortality in different parts starting from May 2009. FINDINGS: The suspected swine flu outbreak from Gwalior India during October- December 2009 was confirmed through S-OIV HA gene specific RT-LAMP and real time RT-PCR. Positive samples through CDC real time and Lamp assay were further processed for isolation of the virus. Full HA gene sequencing of the H1N1 isolates of Gwalior, India revealed 99% homology with California and other circulating novel swine flu viruses. Three major changes were observed at nucleotide level, while two major amino acid shifts were observed at the position C9W and I30M corresponding to the ORF with prototype strain. The HA gene sequence phylogeny revealed the circulation of two genetically distinct lineages belonging to Clade VII and Clade I of S-OIV. CONCLUSIONS: Our findings also supported the earlier report about circulation of mixed genogroups of S-OIV in India. Therefore continuous monitoring of the genetic makeup of this newly emergent virus is essential to understand its evolution within the country. PMID- 21645422 TI - Characterization of a synthetic bacterial self-destruction device for programmed cell death and for recombinant proteins release. AB - BACKGROUND: Bacterial cell lysis is a widely studied mechanism that can be achieved through the intracellular expression of phage native lytic proteins. This mechanism can be exploited for programmed cell death and for gentle cell disruption to release recombinant proteins when in vivo secretion is not feasible. Several genetic parts for cell lysis have been developed and their quantitative characterization is an essential step to enable the engineering of synthetic lytic systems with predictable behavior. RESULTS: Here, a BioBrickTM lysis device present in the Registry of Standard Biological Parts has been quantitatively characterized. Its activity has been measured in E. coli by assembling the device under the control of a well characterized N-3-oxohexanoyl-L homoserine lactone (HSL) -inducible promoter and the transfer function, lysis dynamics, protein release capability and genotypic and phenotypic stability of the device have been evaluated. Finally, its modularity was tested by assembling the device to a different inducible promoter, which can be triggered by heat induction. CONCLUSIONS: The studied device is suitable for recombinant protein release as 96% of the total amount of the intracellular proteins was successfully released into the medium. Furthermore, it has been shown that the device can be assembled to different input devices to trigger cell lysis in response to a user defined signal. For this reason, this lysis device can be a useful tool for the rational design and construction of complex synthetic biological systems composed by biological parts with known and well characterized function. Conversely, the onset of mutants makes this device unsuitable for the programmed cell death of a bacterial population. PMID- 21645423 TI - Undifferentiated connective tissue disease presenting with prevalent interstitial lung disease: case report and review of literature. AB - Undifferentiated connective tissue diseases (UCTDs) are clinical entities characterised by signs and symptoms suggestive of a systemic autoimmune disease, which do not fulfil the diagnostic criteria for a defined connective tissue disease. Lung involvement can complicate the course and management of the disease, often determining a worse outcome. Respiratory dysfunction as the first clinical manifestation has seldom been reported.We describe a case of a female patient who developed significant respiratory dysfunction as the principal clinical sign. Video-assisted thoracoscopy was performed and a histological pattern of nonspecific interstitial pneumonia (NSIP) was found. A pathological diagnosis suggested careful follow-up with extensive immunological screening which then detected Raynaud's phenomenon and positivity of antinuclear antibodies. After a multidisciplinary discussion (pneumologist, radiologist, pathologist and rheumatologist) a final diagnosis of NSIP associated with UCTD was made. The diagnosis of UCTD should be considered when NSIP is diagnosed even in cases with evident first clinical manifestations of severe respiratory dysfunction. A multidisciplinary approach in the field of interstitial lung disease with NSIP, also including rheumatologic expertise, is fundamental to achieve a prompt and correct diagnosis. PMID- 21645424 TI - Relative validity of a food frequency questionnaire for pregnant women. AB - PURPOSE: Pregnant women's diets should be monitored to ensure adequacy, but few studies have assessed the validity of dietary assessment tools among pregnant women. We examined the relative validity of a self-administered, semiquantitative food frequency questionnaire (FFQ) adapted for use in the International Trial of Antioxidants in the Prevention of Preeclampsia for assessing usual diet during pregnancy. METHODS: A subsample (n=107) was recruited for the FFQ validation study, and provided three days of nonconsecutive three-day food records (3D-FRs) following completion of the FFQ. RESULTS: Mean +/- standard deviation (median) energy intakes (kcal/kJ) from the FFQ and mean of 3D-FRs were 1963 +/- 610 (1860)/8219 +/- 2554 (7787) and 2320 +/- 607 (2354)/9713 +/- 2541 (9856), respectively. Spearman correlation coefficients between unadjusted FFQ and 3D-FRs nutrients were positive (rS ranged from 0.17 for iron to 0.49 for folate) and were generally statistically significant (0.05= 40% or to normal) were achieved. In UDCA-monotherapy group, no statistical difference existed in biochemical responses before adding glucocorticoids, whereas the levels of ALT, AST, GLB and IgG decreased significantly when combined with glucocorticoids. No statistical difference of rates of biochemical responses eixted between the two groups, whereas variance could be seen in different pathological stages. Alleviation of inflammatory infiltration after therapy appeared in 3 patients. CONCLUSION: Combination therapy of UDCA with glucocorticoids could be suitable for AIH-PBC overlap syndrome. Early treatment is of benefit for achieving better biochemical response and pathological improvement. PMID- 21645440 TI - [Clinical and pathological features of primary biliary cirrhotic patients with negative anti-mitochondria antibody]. AB - OBJECTIVE: To explore the clinical and pathological features of primary biliary cirrhosis (PBC) patients with negative anti-mitochondria antibody (AMA). METHODS: Two hundreds and eight PBC patients were enrolled. The clinical and histological data of the negative AMA cases were compared with the AMA/AMA-M2 positive cases. RESULTS: 30 out of the 208 cases (14.4%) were AMA negative patients in our study. The general status, biochemical tests and histological findings between the two groups had no significant difference (P > 0.05). The Gamma-globulin, IgG, IgM and IgA levels of AMA/AMA-M2 positive PBC patients were higher than that of the AMA negative cases (P < 0.05). The abnormal rate of cholesterol in AMA negative PBC patients was 65.4% as compared to 50.4% in AMA/AMA-M2 positive cases, no significant difference existed between (P > 0.05). Anti-nuclear antibody (ANA) was observed in 29 (96.7%) AMA negative PBC patients, including 14 (48.3%) with granular pattern, 8 (27.6%) with nuclear membrane pattern, 6 (20.7%) with kinetochore pattern and 1 (3.4%) with homogeneous pattern. AMA negative PBC patients had elevated serum ALP, GGT, IgM and cholesterol levels, and decreased serum AST, IgG and IgA levels as compared with that of autoimmune hepatitis patients (P < 0.05, respectively). CONCLUSION: In cholestatic patients with elevated IgM and cholesterol levels, ANA positive with non-homogeneous pattern, the diagnosis of PBC should be suspected, albeit AMA negative. The clinical, biochemical and histological features of the AMA negative PBC patients were similar to classic PBC patients, but quite different from autoimmune hepatitis. PMID- 21645441 TI - [PD-1/PD-L1 expressions in liver tissues of patients with chronic HBV infection]. AB - OBJECTIVE: To detect and compare the PD-1/PD-L1 (programmed death 1/programmed death 1 ligand) expressions in the liver tissues of chronic HBV infection patients in immune tolerant phase and those in immune clearance phase. METHODS: Liver biopsy samples were divided into two groups: 25 samples from patients in immune clearance phase and 19 samples from patients in immune tolerant phase. PD 1/PD-L1 expressions on T lymphocytes in these liver biopsy specimens were detected by immunohistochemistry method. Percentage of PD-1/PD-L1 positive cells among CD3 positive cells was calculated by semi-quantitative evaluation. Differences between the two groups were statistically analyzed. RESULTS: PD-1/PD L1 expressions were significantly higher in the patients in immune tolerant phase as compared to that in immune active phase (P < 0.05). No statistical difference found between the two groups for PD-L1 expression in Kupffer cells (P > 0.05). CONCLUSION: PD-1/PD-L1 expression level can reflect the immune functions of chronic hepatitis B patients. PMID- 21645442 TI - [Eighty three cases of post transfusion HCV infection: a 10-year follow up]. AB - OBJECTIVE: To find out clinical characteristics and natural history of post transfusion HCV infection. METHODS: 83 subjects who have received the blood from a same blood donor from January 1998 to July 2002 were investigated by the method combining cross-sectional study with retrospective study. HCV-antibody, HCV RNA, liver function, abdomen B-ultrasound and Fibroscan were detected. RESULTS: The HCV-antibody were all positive. The HCV RNA of 56 out of the 83 cases were positive. The chronicity rates of hepatis C were 76.3% (29/38) in male patients and 60.00% (27/45) in female patients respectively, without significant difference (X2 = 2.99, P = 0.11). The average age of the HCV RNA positive patients was (36.54 +/- 14.37) years old. The average age of the HCV RNA negative patients was (27.43 +/- 12.51) years old. A significant difference (T = -2.41, P = 0.018) existed between. The HCV genotype was type1b. Among the HCV RNA positive patients,10 cases were with mild asthenia, anorexia and abdominal distention, 9 cases with increased serum ALT, 12 cases.with chronic hepatitis and 1 case was diagnosed with decompensated liver cirrhosis. CONCLUSION: The clinical manifestations of HCV infection are occult and chronic. The chronicity rate is related to gender and the age when infection was caught. PMID- 21645443 TI - [The clinical study of percutaneous transhepatic radiofrequency ablation combined with tumor edge of percutaneous absolute ethanol injection on liver cancer adjacent to major blood vessels]. AB - OBJECTIVE: To explore the effects of percutaneous transhepatic radiofrequency ablation (PRFA) combined with tumor edge of percutaneous absolute ethanol injection (PEI) on liver cancer adjacent to major blood vessels. METHODS: Seventy five patients with liver cancer adjacent to major blood vessels were randomly divided into two groups: PRFA+PEI therapy group (38 cases) and PRFA control group (37 cases). Tumor necrosis rate, AFP levels, local recurrence rate, median for survival time and cum survival were used as the evaluation index to evaluate the efficacies of the two methods. RESULTS: Tumor necrosis rates of the therapy group and the control group were 84.2% and 54.1% (P < 0.01), respectively; AFP levels of therapy group and control group at 1, 3, 6 and 12 months after treatment were (105.0 +/- 35.5) MUg/L, (28.4 +/- 4.3) MUg/L, (58.6 +/- 6.7) MUg/L, (89.5 +/- 12.5) MUg/L and (137.2 +/- 34.6) MUg/L, (84.2 +/- 18.4) MUg/L, (106.6 +/- 20.3) MUg/L, (173.7 +/- 32.0) MUg/L, respectively. The rates of therapy group was significantly lower than of control group. Local recurrence rates of the therapy group and control group were 2.6%, 7.9%, 13.2% and 31.6% vs 10.8%, 21.6% , 40.5% and 62.1% (P < 0.05) at 3, 6, 12 and 24 months after treatment, respectively. Median for survival time of the therapy group and control group were 28.0 +/- 2.8 months and 19.0 +/- 3.6 months, respectively. Cum survival of the therapy group and control group were 84.2%, 78.9%, 60.5% and 31.6% vs 78.4%, 67.6%, 37.8% and 8.1% (P < 0.05) at 6, 12, 24 and 36 months after treatment, respectively. CONCLUSION: PEI as a supplementary treatment of PRFA can effectively improve the treatment of liver cancer adjacent to major blood vessels and significantly reduce the local recurrence rate and improve long-term survival rates. PMID- 21645444 TI - [Role of mPGES-1 in the occurrence, progression, metastasis and invasion of hepatocellular carcinoma]. AB - OBJECTIVE: To study the expression of mPGES-1 in hepatocellular carcinoma (HCC), observe the effect of MK886 on down-regulation of mPGES-1 gene expression on the biology of human hepatocarcinoma cell line HepG2 and to investigate its significance in the occurrence, progression, metastasis and invasion. METHODS: HCC tissues, para-carcinoma tissues, far-carcinoma tissues and normal liver tissues were collected. The expressions of mPGES-1 were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The proliferation, adherence, migration and invasion abilities of HepG2 cells interfered by MK886 were assessed by MTT and transwell technique respectively. RESULTS: The expression of mPGES-1 in HCCs was higher than that in normal liver tissues (P < 0.01), which increased following histological grade. Furthermore, mPGES-1 expression level was higher in the capsule invasion and metastasis tumor than in primary locus. A significant dose-dependent down-regulation of expressions of mPGES-1 gene mRNA and protein were observed in HepG2 cells when MK886 was given for 48 h (F = 140.402, P < 0.01; a'= 0.00714, P < 0.01). Compared with the control group, the growth inhibitory rate of HepG2 cell was observed significantly time and dose-dependent when MK886 was given. The rate of adhesion cells in experimental groups were 85.3% +/- 1.3%, 70.5% +/- 1.5% and 45.8% +/- 2.4%, respectively, less than that in control group 100.0% +/- 0 (F = 626.313, P < 0.01). The migration cells was 92.47 +/- 1.90, 62.63 +/- 1.96 and 37.33 +/- 0.83 respectively in the experimental groups after 24 h, lower than that in the control group 128.93 +/- 2.60 (F = 1253.805, P < 0.01). The invasion assay revealed that the invading cells were 41.67 +/- 1.30, 25.47 +/- 1.30 and 13.93 +/ 1.66 in the experimental groups, in contrast to 55.67 +/- 2.08 in control group after 24 h. The difference between these groups was significant (F = 372.615, P < 0.01). The numbers of adhesion, migration and invasion of HepG2 cells were dose dependent in MK886 groups. CONCLUSION: Over-expression of mPGES-1 was associated with the tumorigenesis and progression of HCC. The down-regulation of mPGES-1 gene expression might indicated the decrease of the invasion and metastasis of HCC. PMID- 21645445 TI - [General characteristics and clinical practices of Chinese patients with non alcoholic fatty liver disease]. AB - OBJECTIVE: To assess the characteristics and daily treatment compliance of non alcoholic fatty liver disease (NAFLD) patients in China. METHODS: NAFLD adult patients from 21 clinics of 12 cities in China were enrolled in this registry. Physical examination such as demographic characteristics (height, weight, waist circumference measurement), blood pressure and clinical laboratory and ultrasonographic examination of liver were undertaken. Daily practice including life style and medication were recorded and assessed in accordance with 2006 Chinese NAFLD treatment guidelines. RESULTS: A total of 1656 patients were enrolled (1146 male and 510 female), mean of 45.8 +/- 12.6 years old, mean duration of NAFLD history was (47.2 +/- 47.7) months. 44.9% of NAFLD were suffering from metabolic syndromes. Patients with central obesity have higher incidence of hypertension and lower level of high-density lipoprotein cholesterol (HDL-C) than those without central obesity, P < 0.05. Body mass index (BMI), waist circumference, triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) in ALT abnormal group were higher than those in ALT normal group (P < 0.05), HDL-C was lower in ALT abnormal group (P < 0.05). Significant differences existed between the BMI, female waist circumference, TG, fast insulin, HOMA index, ALT, AST and HDL-C among subgroups with mild, moderate and severe steatosis. Majority of the patients did not follow recommendations of NAFLD treatment guidelines. Among targeted population only 15.3% of patients used insulin sensitizers and 23.8% took lipid lowering medicine according to the guideline. CONCLUSION: Data indicated that nearly half of NAFLD patients co morbid with metabolic disorders. Therapy compliance was unsatisfactory and the gap between current practice and Chinese NAFLD treatment guidelines was not optimal. PMID- 21645446 TI - [Effect of ozonized saline on signaling passway of Keap1-Nrf2-ARE in rat hepatocytes]. AB - OBJECTIVE: To study the effect of ozonized saline on the activation of the Keap1 Nrf2-ARE signaling pathway in rat liver cells. METHODS: Twenty male Sprague Dawley rats were randomly divided into ozonized saline (OS) group, model group, ozonized saline control (OSC) group and normal control (NC) group. The rats in OS group and model group were intravenously administered with OS or oxygen saline (5 ml/kg) respectively, once a day for 15 days, and then intraperitoneally injected with CCl4 dissolved in oliver oil. The rats in OSC group were pretreated with OS for 15 days. The rats in NC group were fed normally for 15 days. On the 16th day, the rats in OSC group and NC group were intraperitoneally injected with oliver oil (2 ml/kg) without CCl4. After 24 hours of CCl4 or olive oil intraperitoneal injection, the serum levels of alanine transaminase (ALT) and aspertate aminotransferase (AST) were measured. The liver tissues were also collected for detection of total anti-oxygen capability (TAOC), glutathione (GSH), catalase (CAT), Glutathione peroxidase (GPx). Western Blot was used to detect Nrf2 and immunofluorescence staining assay to display intracelluar distribution of Nrf2. RESULTS: Compared with the rats in model group,the serum ALT and AST levels of rats in OS group were significantly lower (P < 0.01) ,which were (1240.4 +/- 188.2) U/L and (1245.4 +/- 176.9) U/L vs (539.8 +/- 175.3) U/L and (546.0 +/- 130.2) U/L, and the TAOC, CAT, GPx and GSH activity of rats in OS group were significantly higher, which were (0.72 +/- 0.24) U/mg, (1.05 +/- 0.21) mg/g, (676.9 +/- 115.1) U/mg and (45.2 +/- 14.3) U/mg vs (1.37 +/- 0.19) U/mg, (2.23 +/ 0.55) mg/g, (1024.6 +/- 162.9) U/mg and (68.2 +/- 9.9) U/mg, respectively. In contrast with NC group, pretreatment of OS in OSC group elevated TAOC, CAT, GPx and GSH activity (P < 0.01 or P < 0.05). Ozonized saline can strengthen the Nrf2 expression in liver cells. CONCLUSION: Preconditioning injection of ozonized saline can reduce rat's liver injury induced by CCl4. The ozonized saline, as a novel Nrf2 activator, can reduce the oxidative damage of radical oxygen species (ROS) and the deleterious substance by activating the Keap1-Nrf2-ARE signaling pathway and its downstream genes expression. PMID- 21645447 TI - [Effects of Feiyanning on the expression of epithelial-mesenchymal factors in highly metastatic lung cancer cells 95-D]. AB - BACKGROUND AND OBJECTIVE: Our previous in vivo experiment (paper has been published) has confirmed Feiyanning's regulation effect on epithelial cell markers Alpha-catenin, beta-catenin, E-Cadherin and mesenchymal cell markers N cadherin, Fibronectin, vimentin. Based on previous study, the objective of this study is to further investigate the expression of epithelial-mesenchymal cell marker gene and protein intervened by Feiyanning in highly metastatic lung cancer cells 95-D in vitro. METHODS: Human highly metastatic lung cancer 95-D cells were treated with different concentrations of Feiyanning in vitro, and then Real-time PCR and Western blot methods were used to detect mRNA and protein expression of epithelial-mesenchymal cell marker factor Alpha-catenin, beta-catenin, E Cadherin, N-cadherin, Fibronectin and vimentin. RESULTS: Real-time PCR results showed that compared with the control group, Feiyanning at 20% concentration remarkably up-regulated the expression of Alpha-catenin (P<0.05), Feiyanning at 20% and 25% concentrations remarkably up-regulated the expression of E-Cadherin (P<0.05, P<0.01), Feiyanning had no significantly effect on beta-catenin at any concentration, 5% and 10% Feiyanning at 5% and 10% concentrations significantly dow-regulated the expression of mesenchymal cell marker vimentin factor (P<0.01), Feiyanning had no regulatory role on N-cadherin and Fibronectin at any concentration. Western blot results showed that Feiyanning at 5%, 10%, 15%, 20% and 25% concentrations significantly increased E-Cadherin protein expression (P<0.01), but had no regulation effect on Alpha-catenin and beta-catenin protein expression; Feiyanning at 5% and 10% concentrations had a down-regulation effect on N-cadherin and Fibronectin protein expression (P<0.01), and had no regulation effect on vimentin protein expression. CONCLUSION: Feiyanning play a role in inhibiting the adhesion of tumor heterogeneity and the athletic ability by regulating epithelial-mesenchymal cell marker factor expression, and thus inhibit the invasion and metastasis of lung cancer. PMID- 21645448 TI - [The correlation between the up-regulation of Hsp90 and drug resistance to cisplatin in lung cancer cell line]. AB - BACKGROUND AND OBJECTIVE: Hsp90 is a major molecular chaperone, which overexpression is involved in oncogenesis, development and drug resistance in many human cancers. The aim of this study is to investigate the relationship between the GGA-induced overexpression of Hsp90 and chemoresistance to Cisplatin in SPCA-1 and H446 cell line. METHODS: The protein expressions of Hsp90 induced by GGA at different concentrations were analyzed by Immunofluorescence and Western blotting. Cells survival to Cisplatin was determined using the MTT assays. The effect of Hsp90 expression on the drug resistance to Cisplatin in two Cell Lines was analyzed. RESULTS: Compared with the respective control cells, Hsp90 expressions in both experimental cell lines were up-regulated obviously, exhibiting a dose-dependent manner to GGA. MTT assays revealed that the IC50s of cisplatin also showed a substantial elevation for the experimental cells of SPCA 1 and H446, and this elevation was also associated with GGA concerntration. CONCLUSION: GGA is effective for the induction of Hsp90 in the SPCA-1 and H446 cell line. Up-regulated Hsp90 is associated with the chemoresistance to Cisplatin in SPCA-1and H446 cells. PMID- 21645449 TI - [Screening and identification of microRNAs related to acquired gefitinib resistance in lung adenocarcinoma cell lines]. AB - BACKGROUND AND OBJECTIVE: Acquired gefitinib-resistance was closely related to inefficiency of EGFR-TKI treatment in lung adenocarcinoma. However, it was not clear that how microRNAs influenced the acquired gefitinib-resistance in lung adenocarcinoma. The aim of this study is to screen and identify the microRNAs correlated with the acquired gefitinib -resistance in lung adenocarcinoma. METHODS: Morphological difference was observed in gefitinib-sensitive lung adenocarcinoma cell line PC9 cell line and gefitinib-resistance lung adenocarcinoma cell line PC9/AB11 cell line derived from PC9 cell line. Cell cycles and doubling time were detected by flow cytometry, IC50 of gefitinib was evaluated by MTT assay. The differential microRNAs related to acquired gefitinib resistance were screened and identified by microRNA array and real-time PCR. RESULTS: There were obvious morphological differences between PC9 and PC9/AB11 cells. The doubling time, distribution of cell cycle, and the IC50 between PC9 and PC9/AB11 were significantly different. In microarray analysis, compared with PC9 cell line, 4 up-regulated microRNAs were found in PC9/AB11 cells, 9 down regulated microRNAs were found in PC9/AB11 cells. Real-time PCR revealed that miR 138 was significantly down-regulated in PC9/AB11 cells, accord with the microarray. CONCLUSION: MicroRNAs are involved in acquired Gefitinib-resistance of lung adenocarcinoma. Our data presented here to provide an experimental basis and theory thereunder for further study of effect and molecule mechanism underlying the acquired gefitinib-resistance of lung cancer. PMID- 21645450 TI - [Enrichment and function research of large cell lung cancer stem cell-like cells]. AB - BACKGROUND AND OBJECTIVE: There are no universal method to recognize and screen for lung cancer stem cell markers and indicators. Commonly used methods are flow Cytometry and learning from other cancer stem cell sorting tags to sort lung cancer stem cells. But this method has low specificity screening, the workload is huge. In this study, Serum-free suspension culture was used to enrich lung cancer stem cells, and explore method for lung cancer stem cell screening. METHODS: Human large lung cancer cell line-L9981 was cultured in serum-free and growth factors added medium, and spheres were obtained. Then the morphological differences of sphere cells and adherent L9981 cells cultured in serum-containing mediums are observed. Cell proliferation was analyzed by Vi-cell viability analyzer; invasion ability was tested by transwell assay; and in vivo tumorigenicity of the two groups of cells was studied in nude mouse. RESULTS: Compared with adherent L9981 cells cultured in serum-containing mediums, cells cultured in serum-free medium display sphere appearance. Doubling time of adherent cells and sphere cells are (56.05+/-1.95) h and (33.00+/-1.44) h respectively; Spheroid cells had higher invasion and tumorigenicity ability, 5 times and 20 times respectively, than adherent cells. CONCLUSION: Suspension cultured L9981 in Serum-free medium could form spheroid populations. Cells in spheres had higher ability of invasion and Tumorigenicity than adherent L9981 cells. These results indicated spheroid L9981 cells contained enriched lung cancer stem cells, and Serum-free suspension culture can be a candidate method for enriching lung cancer stem cell. PMID- 21645451 TI - [The clinical significance of Claudin-7 and slug expression in lung squamous cell carcinoma and adenocarcinoma]. AB - BACKGROUND AND OBJECTIVE: The aim of this study is to investigate the expression of Claudin-7, Slug and their correltion with clinicopathological characteristics in lung squamous cell carcinoma and adenocarcinoma. We also investigated the relationship between the two factors. METHODS: The expressions of Claudin-7 and Slug in proteins were detected in 101 cases of lung squamous cell carcinoma and adenocarcinoma samples by immunohistochemistry SP method, and Western blot was applied to detect the expressions of Claudin-7 and Slug in 30 specimens of fresh lung cancer and corresponding paracancerous tissues. RESULTS: The expression of Claudin-7 was remarkably decreased in squamous cell carcinoma and adenocarcinoma tissues compared with normal lung tissue, and its expression level was closely correlated with differentiation grade and lymphatic metastasis (P<0.05), whereas the expression of Slug was significantly higher in cancer tissues than that in normal lung tissue. Outside differentiation grade and lymphatic metastasis, the expression of Slug was related to TNM stage (P<0.05). The expression of Claudin-7 was negatively correlated with the expression of Slug in lung squamous cell carcinoma and adenocarcinoma (r=-0.566,8). CONCLUSION: The down-regulation of Claudin-7 and overexpression of Slug might be one of pertinent biological markers for malignant transformation and metastasis of lung squamous cell carcinoma and adenocarcinoma. PMID- 21645452 TI - [The clinical significance of beta-arrestin 2 expression in the serum of non small cell lung cancer patients]. AB - BACKGROUND AND OBJECTIVE: Non-small cell lung cancer (NSCLC) with high morbidity and mortality is the most common types of lung cancer. beta-arrestin 2 is a kind of soluble protein regulating signal transduction mediated by G protein coupling receptor. The aim of this research is to evaluate the clinical significance of beta-arrestin 2 expression in the serum of NSCLC patients. METHODS: The clinical and follow-up data of 20 healthy candidates and 67 patients diagnosed with NSCLC in Sun Yat-sen University Cancer Center from January 2005 to December 2006 was retrospectively analyzed. ELISA was applied to detect the expression of beta arrestin 2. RESULTS: The serum level of beta-arrestin 2 in NSCLC patients were all Significantly lower than those in healthy controls (P<0.001, P<0.001, P<0.001). The serum level of beta-arrestin 2 in stage I NSCLC patients were higher than those in stage III as well as in stage IV (P<0.001, P<0.001). No statistical difference of beta-arrestin 2' serum level was found between with stage III and stage IV patients (P=0.273). Univariate prognostic factor analyzed by Kaplan-Meier method indicated patients' prognosis with high serum level of beta-arrestin 2 was better than patients with low and middle (P<0.001, P<0.001). The serum level of beta-arrestin 2 and the stage of NSCLC signally affected prognosis in COX regression model (P=0.003, P=0.004). CONCLUSION: The serum level of beta-arrestin 2 had significant difference between NSCLC patients and healthy controls, likewise between the early and advanced NSCLC patients. The serum level of beta-arrestin 2 affected NSCLC patients' prognosis. PMID- 21645453 TI - [Prognostic factors in 408 elderly lung cancer patients more than 70 years old]. AB - BACKGROUND AND OBJECTIVE: With the aging of the population, the incidence of lung cancer in elderly rises. It showed that the incidence and mortality of lung cancer in people over 70 years were increased in the past 10 years. We defined age 70 as boundary line of the elderly patients in lung cancer. The aim of this study is to analyze the factors of affecting prognosis. METHODS: A retrospective study had enrolled 408 cases of lung cancer aged over 70 and SPSS 13.0 software was used in univariate analysis and COX regression analysis to analyze factors affecting prognosis, such as gender, age, complications, symptoms, pathological type, clinical stage, effusion, surgery, radiotherapy, chemotherapy and so on. RESULTS: In univariate analysis, symptoms, stage, effusion, surgery, chemotherapy and chemotherapy cycles showed affecting prognosis significantly. In COX regression analysis, it showed that clinical stage (P<0.001), surgery (P=0.013), chemotherapy cycles (P=0.001) were independent prognostic factors. CONCLUSION: Elderly lung cancer patients could be benefit from surgery and adjuvant chemotherapy while early stage. At late stage, their survival time may be prolonged when receive chemotherapy at least 4 cycles. Single-agent chemotherapy would be a good choice for elderly lung cancer. Effusion, particularly, pericardial effusion significantly influenced the prognosis, so that it should be effectively controlled. PMID- 21645454 TI - [Study on point mutations of K-ras gene in non-small cell lung cancer in Guangxi]. AB - BACKGROUND AND OBJECTIVE: Recent studies indicated that non-small cell lung cancer (NSCLC) patients with mutant K-ras were resistant to epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs). The aim of this study is to explore the relationship between the mutation of K-ras gene and NSCLC in Guangxi by detecting the point mutations in codon 12, 13 and 61 of K-ras gene in NSCLC. METHODS: The point mutations in codon 12, 13 and 61 of K-ras gene were detected by single-strand conformation polymorphism (SSCP) analysis of polymerase chain reaction (PCR) products and DNA sequencing analysis in 105 cases of NSCLC tissues and 30 cases of adjacent normal tissues. RESULTS: No point mutation in codon 12, 13 and 61 of K-ras gene was found in 105 cases of NSCLC tissues and 30 cases of adjacent normal tissues. In this study, the mutation frequency of K-ras gene in NSCLC was 0 (0/105). CONCLUSION: The high proportion of K-ras gene in wild-type indicates that patients with NSCLC in Guangxi could take more benefits from the therapy with EGFR-TKIs. PMID- 21645455 TI - [Expressions of c-Cbl, Cbl-b and EGFR and its role of prognosis in NSCLC]. AB - BACKGROUND AND OBJECTIVE: Epidermal growth factor receptor (EGFR) is closely correlated with the progression of lung cancer. Its activity is modulated by Casitas B-lineage lymphoma (Cbl) family. The aim of this study is to investigate the expression and clinical relevance of c-Cbl, Cbl-b and EGFR in non-small cell lung cancer (NSCLC). METHODS: Expressions of c-Cbl, Cbl-b and EGFR protein were detected with tissue microarrays and immunohistochemistry technique in 94 cases of NSCLC. The correlations between the expression of the three proteins and clinicopathological parameters were analyzed. RESULTS: The positive expression rates of EGFR, c-Cbl and Cbl-b were 60.6% (57/94), 30.9% (29/94) and 84.0% (79/94), respectively. The expression of EGFR, c-Cbl and Cbl-b was not associated with age, pathological type, TNM stage, lymph node metastasis, and smoking history. c-Cbl and Cbl-b status was not significantly correlated with overall survival. Subgroup analyses showed that c-Cbl-positive patients had longer survival than c-Cbl-negative patients in EGFR-positive group (P=0.014). CONCLUSION: Detection of c-Cbl protein levels might contribute to the prognosis evaluation of EGFR-positive NSCLC. PMID- 21645456 TI - [Discordance of epidermal growth factor receptor mutations between primary and corresponding metastatic tumors in non-small cell lung cancer]. AB - BACKGROUND AND OBJECTIVE: Epidermal growth factor receptor (EGFR) gene mutation assay has been applied to select the chemosensitive patients for tyrosine kinase inhibitor (TKI) treatment widely. The aim of this study is to determine the discordance of EGFR mutations between primary and corresponding metastatic tumors in non-small cell lung cancer (NSCLC). METHODS: Thirty five paired primary tumors and corresponding metastases from Cancer Center of Sun Yatsen University were evaluated for the EGFR mutations by TaqMan RT-PCR analysis. RESULTS: EGFR mutations were detected in 29 of 35 primary tumors and in 18 of 35 corresponding metastases. 31.43% (11 of 35, P=0.008) showed discordance in EGFR mutations between primary tumors and corresponding metastases. The EGFR mutation status was consistent in 68.57% (24 of 35) patients. CONCLUSION: EGFR mutations were discordant between the primary tumor and the corresponding metastases in NSCLC. PMID- 21645457 TI - [The common causes of conversion of VATS during operation for 248 non-small cell lung cancers]. AB - BACKGROUND AND OBJECTIVE: VATS-lobectomy has been used as regular surgical procedure clinically for non-small cell lung cancer. The aim of this study is to evaluate the surgical emergencies during VATS and the related factors postoperatively. METHODS: Clinical data were reviewed for patients who were performed with pulmonary related surgery between January 2006 and July 2008 in our department. RESULTS: 248 (117 CVATS and 131 AVATS) VATS lobectomy were performed, including 13 cases that were transferred into AVATS or OPEN. The common related reasons were bleeding of pulmonary branches, adhesion, anatomic deformity, bleeding of azygos and bleeding of middle-lobe-vein. 129 thoracotomy cases were enrolled. Compared with OPEN surgery, VATS got the merits of short in hospital duration (20 days vs 27 days, P=0.015), less bleeding (197 mL vs 250 mL, P=0.005) and less pain (4.6 vs 6.2, P=0.003). CONCLUSION: VATS is a safe surgical procedure for early stage NSCLC with merits of lower morbidity and sooner recovery. So it could be concluded that in some circumstances, VATS could be chosen as an alternate of thoractomy. PMID- 21645458 TI - [Role of erlotinib 100 mg/d in the treatment of advanced adenocarcinoma lung cancer failed to previous chemotherapy]. AB - BACKGROUND AND OBJECTIVE: Erlotinib is a epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI). The standard dose is 150 mg/d. But some patients have to reduce the dose or stop treatment due to its side effects. The aim of this study is to evaluate the efficacy and safety of erlotinib 100 mg on patients with advanced aden carcinoma lung cancer previously failed to chemotherapy. METHODS: Forty adenocacinoma patients with histologically confirmed diagnosis were enrolled to be given erlotinib 100 mg/d until disease progression or occurrence of intolerable toxicity. Relationships between therapeutic effects of erlotinib, progression-free survival, side effects and the clinical characters were analyzed. RESULTS: Among 40 patients, 11 got partial response, 14 had a stable disease and 15 had a progressive disease. The total response rate was 27.5%, and the disease control rate was 62.5%. The median progression-free survival was 5.5 months. Fifteen patients were positive for EGFR mutation according to the result of peripheral blood test. The response rate and disease control rate in 15 EGFR-mutation-positive patients were 33.3% and 73.3%, respectively, and the PFS was 8.4 months for 15 patients. The patients with ECOG 0-1 had a significant better PFS than those with ECOG 2 (6.1 vs 3.1 months) (P=0.043). The most frequent toxicities of erlotinib were rash and diarrhea which were easily controlled. CONCLUSION: Erlotinib 100 mg is effective and safe in treatment of patients with advanced aden carcinoma, especially for non-smoking patients, patients with good performance status and EGFR-mutation-positive patients. PMID- 21645459 TI - [Pre-operative plasma D-dimer level may predict the poor prognosis within one year after the surgery for non-small cell lung cancer]. AB - BACKGROUND AND OBJECTIVE: Some operable non-small cell lung cancer (NSCLC) patients have poor prognosis shortly after the surgery. D-dimer (DD) is an independent prognosis factor of lung cancer, especially for inoperable patients. The aim of the study is to investigate whether the pre-operative plasma DD level could predict the poor prognosis shortly after the surgery in operable NSCLC patients. METHODS: The pre-operative plasma DD level of 56 newly diagnosed NSCLC patients without metastasis was examined. All the patients had been followed for one year post-operatively and the end-point was the occurrence of the poor prognosis incident including any sign of the metastasis, local recurrence or death related with the lung cancer. Difference of prognosis according to pre operative plasma DD level was compared by Chi-square test. Diseases progress was analyzed by Kaplan-Meier method. RESULTS: Among 56 NSCLC patients, 91% had received the curative resections (44 lobectomy and 7 pneumonectomy). There were still 2 cases of the wedge resection and 3 cases of the exploration. The median of the pre-operative plasma DD level was 1.05 (0.55) mg/L. The patients were allocated into two subgroups by the median of the DD levels. There were 11 patients with poor prognosis within one year after the resection in the high DD subgroup, while 3 patients in the low DD subgroup (P=0.03, OR=4.89, 95%CI: 1.2 20.1). The diseases progress curves were significantly different between the high and low subgroups (P=0.024). Based on plasma DD level, the poor prognosis incident within one year after the surgery was best predicted in the early stage (I, II) of the NSCLC, especially in adenocarcinoma patients. CONCLUSION: The pre operative plasma DD levels may predict the poor prognosis within one year after the surgery in NSCLC. The measurement of the fibrinolysis marker may help to exclude the unfit patients for the surgery. PMID- 21645460 TI - [A new target in non-small cell lung cancer: EML4-ALK fusion gene]. AB - It was only 3 years ago that the fusion gene between echinoderm microtubule associated protein-like4 (EML4) and anaplastic lymphoma kinase (ALK) has been identified in a subset of non-small cell lung cancer (NSCLC). EML4-ALK is most often detected in never smokers with lung adenocarcinoma and has unique pathologic features. EML4-ALK fusion gene is oncogenic, which could be suppressed by ALK-inhibitor through blocking the downstream signaling passway of EML4-ALK. This review will focus on the molecular structure, function, biology, detection method and the diagnostic and therapeutic meaning of EML4-ALK of lung cancer. PMID- 21645461 TI - [Advance of circulating tumor cell in patients with lung cancer]. PMID- 21645462 TI - [NP protocol taken for pulmonary sarcomatoid carcinoma: a case report]. PMID- 21645463 TI - [Gemcitabine in RRM1-negative advanced refractory non-small cell lung cancer (NSCLC): a case report]. PMID- 21645464 TI - [A case report of primary lung cancer with chief complaint of dysphagia]. PMID- 21645465 TI - [Primary pulmonary non-Hodgkin's lymphoma: a case report and literature review]. AB - Primary pulmonary non-Hodgkin's lymphoma (PPNHL) is a kind of malignant lymphoma originated from lymphoid tissue of the lung. PPNHL is a rare type in extra nodal lymphoma. In this article, we reported one case of 57-year-old woman with PPNHL, whose clinical characteristics, radiological features, diagnosis and treatment was described and related literatures were reviewed. PMID- 21645466 TI - Responding to rural health needs through community participation: addressing the concerns of children and young adults. AB - A small rural health service undertook a major needs analysis in 2008 to identify gaps in service delivery and duplication of services. This exercise was intended to inform strategic direction but the result was consumer and community consultation and outcomes that far exceeded everyone's expectations. Organisations often pay lip service to the concept of community participation and consultation and the importance of consumer involvement. Turning this rhetoric into action is challenging and requires dedicated staff, organisational support and momentum for it to occur. The project described resulted in targeted, purposeful action regarding community engagement, and the findings and outcomes are reflective of this. The unexpected findings required an organisational shift, which was embraced by the health service and resulted in collaborative partnerships with consumers and organisations that are proving beneficial to the entire community and outlying areas. Few organisations would demonstrate the willingness to accommodate such change, or undertake a needs analysis that is chiefly community driven. PMID- 21645467 TI - A local study of costs for private allied health in Australian primary health care: variability and policy implications. AB - Multidisciplinary approaches to primary health care improve outcomes for individuals living with chronic conditions. However, emerging evidence suggests access to allied health professionals in Australia is problematic. This paper reports findings of a telephone survey of allied health professionals' billing practices in one urban area. The survey was undertaken as a quality improvement project in response to the affordability queries raised by patients and carers in the clinical setting. The aim was to determine financial cost of access to allied health professionals in one urban primary health care setting. Participant practices included: physiotherapy (n=21), podiatry (n=8) and dietitians (n=3). Fees were variable, with cost of the initial (assessment) appointment higher than subsequent (follow-up) appointments in 92% of practices. The average out of pocket expenses for assessment and three follow-up appointments ranged from $258 to $302. When available, the Medicare rebate reduced this to $58-106. Bulk billing was not offered. Variable costs, minimal concessions and absence of bulk billing in this confined geographical area creates a cost barrier to access for patients from lower socioeconomic groups and has implications for access to multidisciplinary care in Australian primary health care. PMID- 21645468 TI - Nutrition and physical activity guidance for women in the pre- and post-natal period: a continuing education needs assessment in primary health care. AB - The study objective was to assess primary health care (PHC) providers' exposure to women in the pre- and post-natal period, current nutrition and physical activity guidance practices, confidence and perceived needs for continuing education relevant to nutrition and physical activity guidance in the peri-natal period. A self-administered cross-sectional questionnaire survey amongst a purposively recruited sample of 226 local primary health care providers evenly distributed across general practice, community nursing, pharmacist and pharmacy assistant worker groups. The questionnaire contained 106 items about primary health care providers' exposure to women in the pre- and post-natal life-stage, their current nutrition and physical activity guidance practices, confidence and perceived needs for continuing education relevant to nutrition and physical activity guidance in the peri-natal period. Results indicate that PHC providers across general practice, community nursing and pharmacy service settings are frequently accessed by women during this life-stage, and regularly and variably provide guidance on nutrition and physical activity, and report different continuing education needs. Continuing education interventions need to be tailored to match the needs of each PHC group. Pharmacy-based staff are a priority for PHC continuing education about nutrition and physical activity if the potential of the community-based pharmacy as a primary health setting is to be realised. PMID- 21645469 TI - Integration of complementary and alternative medicine information and advice in chronic disease management guidelines. AB - The growing evidence on the benefits and risks of complementary and alternative medicine (CAM) and its high rate of use (69% of Australians) - particularly for chronic or recurrent conditions - means increasing attention on CAM. However, few people disclose CAM use to their GP, and health professionals tend to inadequately discuss CAM-related issues with their patients, partly due to insufficient knowledge. As clinical and non-clinical chronic condition management guidelines are a means to educate primary health care practitioners, we undertook a content analysis of guidelines relevant to two common chronic conditions - cardiovascular disease (CVD) and type 2 diabetes mellitus (T2DM) - to assess their provision of CAM-related information. Ten current Australian guidelines were reviewed, revealing scant CAM content. When available, the CAM-relevant information was brief, in some cases unclear, inconclusive and lacking in direction to the GP or health care provider. Although we focus on CVD and T2DM, we argue the value of all chronic condition management guidelines integrating relevant evidence-informed information and advice on CAM risks, benefits and referrals, to increase GP awareness and knowledge of appropriate CAM therapies, and potentially to facilitate doctor-client discussion about CAM. PMID- 21645470 TI - Importance of effective collaboration between health professionals for the facilitation of optimal community diabetes care. AB - Diabetes places a significant burden on the individuals concerned, their families and society as a whole. The debilitating sequelae of diabetes can be limited or prevented altogether through strict glycaemic control. Despite the seemingly uncomplicated nature of the disorder, effective management can be elusive, as the impact of having to deal with diabetes on a daily basis can be profound and appropriate professional support is not always readily available. As the roles of general practitioners (GPs) and allied health professionals have evolved, a major issue now facing all is that of developing and maintaining effective collaborative relationships for the facilitation of optimal community diabetes care. Using a simple survey methodology, the present exploratory study investigated the referral patterns of GPs to diabetic educators (DEs) working for a community health service in an Australian town, and reasons for referral and non-referral in order to identify factors that contribute to a sound and sustainable collaborative relationship. The results provide some evidence that GPs and DEs in this town do work collaboratively towards achieving client-centred goals and highlight the need to inform GPs who are new to communities, such as this one, of the available DE services. Most importantly, the study identified that there are many opportunities to strengthen collaboration so as to facilitate optimal community diabetes care. This information is valuable, because there is limited empirical evidence either nationally or internationally about the process of collaboration between health professionals in the management of chronic diseases, such as diabetes. PMID- 21645471 TI - Barriers to recruitment of professionals into a general practice childhood obesity program. AB - Recruiting general practitioners to take part in research in primary care is important and challenging. This paper describes the process, barriers and achievements experienced by a research team whilst recruiting Divisions and general practitioners (GPs)/practices into a project related to management of obesity in children, which used Breakthrough Series methodology in the intervention arm. The research team chose to recruit GPs through Divisions of General Practice, because of the existing positive relationship between the research team and the Divisions, and the Divisions' positive relationships with the GPs/practices in their area. The project aimed to recruit four Divisions of General Practice, who in turn would recruit a total of 24 GPs/practices. Four Divisions (two rural, two metropolitan) were recruited initially, but the two rural Divisions withdrew before the project commenced and were replaced by two metropolitan Divisions. As Divisions were unable to recruit sufficient GPs/practices, two additional strategies were used, but despite all efforts, only 17 practices were recruited. Reflection on recruitment failure focussed on reasons from the perspective of Divisions, GPs, other projects using the Breakthrough Series methodology with a chronic disease focus and other projects conducted in Australia where the focus was childhood obesity. The results showed that even Divisions who were enthusiastic about joining a project may be unable to recruit sufficient GPs and practices, for reasons including staff changeover and GP reluctance. Caution is suggested when setting recruitment targets where the condition is sensitive, is not the presenting problem, is not routinely tackled in general practice, involves minors, where treatment is perceived to be of doubtful effectiveness, or where major government policies may need to be considered. PMID- 21645472 TI - National Health and Hospital Reform Commission final report and patient-centred suggestions for reform. AB - The final report of the National Health and Hospital Reform Commission (NHHRC) called for a strengthened consumer voice and empowerment. This has salience for the development of health policy concerning chronic illnesses. This paper compares the recommendations for chronic illness care made in the NHHRC final report with suggestions made by people with chronic illness and family carers of people with chronic illness in a recent Australian study. Sixty-six participants were interviewed in a qualitative research project of the Serious and Continuing Illness Policy and Practice Study (SCIPPS). Participants were people with type II diabetes mellitus, chronic obstructive pulmonary disease or chronic heart failure. Family carers were also interviewed. Content analysis was undertaken and participants' recommendations for improving care were compared with those proposed in the NHHRC final report. Many suggestions from the participants of the SCIPPS qualitative research project appeared in the NHHRC final report, including the need to improve care coordination, health literacy and the experience of Indigenous Australians. The research project also identified important issues of family carers, immigrants and people with multiple illnesses, which were not addressed in the NHHRC final report. More specific attention is needed in health reform to improve the experience of family carers, Indigenous peoples, immigrants to Australia and people with multiple illnesses. To align more closely with their needs, health reform must be explicitly informed by the voices of people with chronic illness and their family carers. The NHHRC recommendations must be supplemented with proposals that address the needs of these people for support and the problems associated with poor care coordination. PMID- 21645473 TI - Parents's plans to supply their adolescents with alcohol. AB - The aim was to determine the extent to which parent and adolescent characteristics and patterns of alcohol use influence parents' plans to supply their adolescent aged 14-16 years with full serves of alcohol (i.e. not necessarily initiation) in the next 6 months. A cross-sectional sample of parents from Victoria, Australia, completed an online survey. Parents' plans to supply alcohol in the next 6 months was significantly associated with their reports of supplying alcohol in the previous 3 months (beta=0.51, P<0.01), perceptions that their adolescent drinks (beta=0.34, P<0.01), reports of not practising religion (beta=0.13, P<0.01), and Alcohol Use Disorders Identification Test scores (beta=0.09, P=0.04). The total variance explained by the model was 57.4% F (9, 242)=36.2, P<0.01. Parents' plans to supply their adolescent with alcohol might be a reflection of the normalisation of alcohol use in Australia. There is a need to support Australian parents to review their own alcohol use, clarify their views on alcohol use by their adolescent and confidently restrict their child's access to alcohol, irrespective of their own drinking patterns. PMID- 21645474 TI - A novel primary care clinical prediction rule for early detection of osteoporosis. AB - The effects of osteoporosis (OP) can be significantly slowed if disease is detected early. We report on a clinical risk prediction rule developed from patient histories taken in an orthopaedic outpatient clinic, before confirmatory testing for OP. Data were extracted from routine audits of consecutive records of patients with recent wrist fracture, comprising demographic details, medications, past and current disease, and fracture details. Clinical prediction rule elements were tested against clinical suspicion of OP. The clinical prediction elements comprised sex and age risk, medications that predispose patients to OP and/or falls, previous fractures and disease/medical conditions that are known OP risks. The best cut point (6.5) demonstrated 100% sensitivity with clinical suspicion of OP. Patient history information is often available before OP is clinically suspected or a definitive diagnosis is made. Our clinical prediction rule will be useful in primary care settings where objective measures of bone health are not readily available. It will raise OP awareness amongst health care providers and patients, particularly those not previously suspected of having OP. It will assist in identifying at-risk patients early and commencing them on appropriate management, without waiting for definitive bone health tests. PMID- 21645475 TI - 'Perfect skin', the media and patients with skin disease: a qualitative study of patients with acne, psoriasis and atopic eczema. AB - The relationship of skin disease with societal ideals of beauty, and the role of the media in this relationship, has not previously been researched. The overall objective of this study was to explore the psychological effects of skin disease. The theme of the ideal of perfect skin and the role of the media in generating this ideal arose via an inductive study methodology and was explored in the context of respondents' psychological morbidity. A qualitative study, 62 semi structured interviews were conducted with respondents with acne, eczema or psoriasis recruited from both general practice and specialist dermatology practice in an Australian regional city. Interviews were audiotaped, transcribed and subjected to thematic analysis employing a process of constant comparison in which data collection and analysis were cumulative and concurrent. The themes of perfect skin, societal ideals and media influence emerged from this iterative process. Respondents identified a societal ideal of flawless skin, largely mediated by media portrayals of perfection. Failure to meet this ideal precipitated psychological morbidity in female, but not male, respondents. An appreciation of the pervasive pressures of society and media upon females with skin disease may inform management strategies, particularly psychological management strategies, in patients with skin disease. PMID- 21645476 TI - Improving the professional support for parents of young infants. AB - The objective of this study was to discuss ideas for improving child health services on the basis of findings of an observational study that was designed to explore the role of child health nurses in supporting parents during the first 6 months following the birth of an infant. As part of a larger study in a child health service in urban Australia, surveys were used to collect data from two independent samples of both parents and nurses at an 8-month interval. Data were condensed using factor analysis; regression analyses were used to determine which aspects of care were most important for the parents, and importance-performance analysis was used to determine which aspects of care needed improvement. While the majority of parents valued support from child health nurses, a need for improvement was identified in empowering parents to make their own decisions, discussing emotional issues with parents, providing continuity of care and giving consistent advice. Organisations should value and provide support for child health nurses in their invisible, non-quantifiable work of supporting families. The structure of child health services should also provide child health nurses continuity of care with the families they support. PMID- 21645477 TI - Culturally and linguistically diverse peoples' knowledge of accessibility and utilisation of health services: exploring the need for improvement in health service delivery. AB - With 28% of Australia's population having a culturally and linguistically diverse (CALD) background, the health system faces an increasing challenge to provide accessible and culturally competent health care. The view that all CALD communities are homogenous and solutions can be developed for the entire nation is detrimental. Despite available health services, CALD communities are reluctant to use them due to cultural differences, perceived racism and misunderstandings leading to the existing health disparities. Therefore, gathering data from four prominent CALD communities, such as the Sudanese, Afghani, Pacific Islander and Burmese communities in Logan, Queensland, about how they perceive and use health services can provide insightful information towards development of a service model that will better suit these CALD communities. The objective of the study was to examine the extent to which four prominent CALD communities (Sudanese, Afghani, Pacific Islander and Burmese) access and use health services in Logan, Queensland. Six focus group interviews using interpreters were conducted in English with Sudanese, Afghani, Pacific Islander and Burmese people. The results indicated that even long-standing CALD communities, such as the Pacific Islander people, were unfamiliar with health services and experienced difficulties accessing appropriate health care. Most wanted doctors to use traditional healing methods alongside orthodox medicine, but did not feel respected for their beliefs. Language difficulties impeded communication with health professionals who were hindered by ineffective use of interpreters. In conclusion, a clear role for bilingual community-based navigators was identified by CALD participants to address concerns about the health system, and to improve accessibility and health service usage. PMID- 21645478 TI - Nutrition advice in general practice: the role of general practitioners and practice nurses. AB - General practice is an ideal setting to be providing nutrition advice; however, it is important that the role of general practitioners (GPs) and practice nurses in providing nutrition advice is acknowledged and defined. This article aims to discuss the role of GPs and practice nurses in the delivery of nutrition advice. Ten general practitioners and 12 practice nurses from a NSW urban Division of General Practice participated in questionnaires and a Lifescripts implementation study, as well as their consenting patients receiving Lifescripts (n=13). An online survey was conducted with 90 Australian private practice dietitians. Semi structured telephone interviews were conducted with 52 Australian private practice dietitians. The provision of basic nutrition advice is acknowledged to be part of the role of GPs and practice nurses, as they are the first point of contact for patients, allowing them to raise nutrition awareness. However, it is important that this advice is evidence based and able to be delivered in a time efficient manner. Increased nutrition education and the availability of appropriate resources and nutrition-related best practice guidelines will assist in this process. PMID- 21645479 TI - [Blood transfusion to non-bleeding, critically ill patients]. PMID- 21645480 TI - [The evidence for blood cell transfusions to non-bleeding, critically ill patients is sparse]. AB - Critically ill non-bleeding patients often receive red blood cell transfusions in the intensive care unit because of anaemia. The evidence that transfusion leads to improved outcome is limited and the treatment may be harmful to some of these patients. Current recommendations support a restrictive transfusion strategy. Additional randomised clinical trials are needed to elucidate the association between specific haemoglobin levels, transfusion and morbidity and mortality in critically ill patients including those with severe sepsis and acute coronary syndrome. PMID- 21645481 TI - [Oxygen therapy in exercise-induced hypoxemia improves physical capacity and reduces dyspnea in patients with COPD]. AB - Many chronic obstructive pulmonary disease patients suffer from dyspnoea and reduced physical abilities. Some of the patients exhibit desaturation during physical exercise. Few studies have examined if oxygen therapy during physical exercise can relieve symptoms and improve outcomes of pulmonary rehabilitation. These studies show that oxygen therapy during exercise results in significant improvement in exercise capacity. Yet it is not possible to draw a clear conclusion regarding the effects of more prolonged (weeks) treatment. The studies are in general small and of poor methodological quality. PMID- 21645482 TI - [Continuous glucose monitoring in pregnancies complicated by diabetes]. AB - In this review the use of continuous glucose monitoring (CGM) in pregnancy complicated by type 1 and type 2 diabetes is examined. Fourteen relevant articles were identified. Observational studies demonstrated that CGM was feasible during pregnancy without severe side effects. One randomised controlled trial resulted in improved metabolic control and reduced risk of macrosomia in women randomised to CGM. Future studies on the use of CGM in pregnancy are awaited with interest. PMID- 21645483 TI - [Introduction of electronic patient record in an emergency department]. AB - The speciality of emergency medicine is a fairly new concept in the Danish hospital setting, and recent years have seen the emergence of many new emergency departments all over Denmark. We here report the conception and implementation of a novel electronic charting program, comprising a combination of text and check boxes, at Holbaek and Koge emergency departments in Region Zealand. We discuss the advantages and drawbacks of electronic charting compared with common tape dictation and discuss how the electronic charting program may be improved to the benefit of physicians and patients alike. PMID- 21645484 TI - [The effect of calcium channel blockers for hypertension--a survey of a Cochrane review]. AB - The Cochrane analysis comprises data from 18 trials with a total of 140,000 patients. Calcium channel blockers (CCBs) were superior to beta-blockers on stroke reduction. CCBs were also superior to angiotensin-converting enzyme inhibitors and angiotensin receptor blockers on stroke reduction, most likely explained by differences in achieved blood pressure. We question the conclusion, that diuretics are superior to CCBs as agents of first choice, but the analysis supports the view taken by many guidelines, that beta-blockers no longer are drugs of first choice. PMID- 21645485 TI - [Tuberculous coxitis in the hips 55 years after primary tuberculosis]. AB - We describe a case concerning tuberculous coxitis 55 years after primary tuberculosis. A 76-year old man developed hip pain, weight loss and a gluteal tumor. After three months he was diagnosed with tuberculous coxitis by ultrasound assisted aspiration from the joint. Anti-tuberculous chemotherapy was commenced. After three months a Girdlestone operation was performed and seven months later a total hip replacement was performed. Anti-tuberculous chemotherapy was continued for a total of 12 months. Early ultrasound assisted aspiration from the joint is recommended to shorten diagnostic delay and optimise treatment of tuberculous coxitis. PMID- 21645486 TI - [Tibial tuberosity traction may cause fractures]. AB - Today, traction pins are often applied during treatment of corpus femoris fractures. Common complications are infection and cut-out of the pin. Fractures through the traction-pin site are an uncommon complication. We describe the case of a 46 year-old male with a horizontal low-energy fracture through the pin site occurring five and a half months after removal of the traction pin. PMID- 21645487 TI - [Splenosis around the internal genitalia--a rare late complication following trauma to the spleen]. AB - A 40 year-old woman underwent abdominal hysterectomy due to hypermenorrhoea. During the operation, several soft tumorous masses were discovered in the cul-de sac and samples were taken. Histological analyses revealed splenic tissue. The patient had a splenectomy during her childhood due to a traumatic spleen rupture. Clinical check-up one year later showed no progression of the masses. Splenosis is a benign condition that frequently occurs after traumatic spleen rupture. It is often asymptomatic and found by coincidence, but as it can mimic malignancy, it often leads to an extensive workup before the diagnosis is established. PMID- 21645488 TI - [Prepyloric antral web--a rare cause of gastric outlet obstruction]. AB - A case of gastric outlet obstruction secondary to prepyloric antral web in a four year-old boy with cerebral pareses is reported. Routine roentgenographic examination was initially misinterpreted as duodenal obstruction. Prepyloric antral web was suspected by subsequent endoscopy and was confirmed by operation. The patient underwent antropyloroplasty that resulted in excellent recovery. The possibility of a prepyloric antral web should be considered in any infant or child with persistent vomiting when pyloric stenosis is excluded. PMID- 21645489 TI - [Picture of the month: hypertrophic pyloric stenosis]. PMID- 21645490 TI - Advanced procedures for labeling of antibodies with quantum dots. AB - Semiconductor quantum dots (QDs) are proved to be unique fluorescent labels providing excellent possibilities for high-throughput detection and diagnostics. To explore in full QDs' advantages in brightness, photostability, large Stokes shift, and tunability by size fluorescence emission, they should be rendered stable in biological fluids and tagged with the target-specific capture molecules. Ideal QD-based nanoprobes should not exceed 15nm in diameter and should contain on their surface multiple copies of homogeneously oriented highly active affinity molecules, for example, antibodies (Abs). Direct conjugation of QDs with the Abs through cross-linking of QDs' amines with the sulfhydryl groups issued from the reduced Abs' disulfide bonds is the common technique. However, this procedure often generates conjugates in which the number of functionally active Abs on the surface of QDs does not always conform to expectations and is often low. Here we have developed an advanced procedure with the optimized critical steps of Ab reduction, affinity purification, and QD-Ab conjugation. We succeeded in reducing the Abs in such a way that the reduction reaction yields highly functional, partially cleaved, 75-kDa heavy-light Ab fragments. Affinity purification of these Ab fragments followed by their tagging with the QDs generates QD-Ab conjugates with largely improved functionality compared with those produced according to the standard procedures. The developed approach can be extended to conjugation of any type of Ab with different semiconductor, noble metal, or magnetic nanocrystals. PMID- 21645491 TI - Near-infrared-labeled tetracycline derivative is an effective marker of bone deposition in mice. AB - Bone-specific compounds have been used effectively for the detection of bone mineralization, growth, and morphological changes. These agents typically contain iminodiacetic acid groups that can form complexes with apatite and fluoresce in the visible spectrum. We exploited a subset of these chemical chelators to produce a near-infrared (NIR) optical bone marker for preclinical animal imaging. By conjugating target compounds to IRDye 800CW, we extended the effective fluorescence signal detection to the NIR region without affecting the compound's ability to function as a marker of the mineralization process. Calcein and a tetracycline derivative (BoneTag agent [BT]) bound specifically to differentiated mineralized osteoblast cultures, with the latter exhibiting 6-fold higher signal intensities. Subsequent in vivo testing demonstrated effective skeletal labeling with IRDye 800CW BT. We were able to identify a changing mineralization front in bone sections from (i) normal growing mice injected with IRDye 800CW BT 6weeks prior to the administration of IRDye 680 BT and (ii) an osteoporosis mouse model comparing cortical bone in sham-treated and ovariectomized mice. These results provide evidence that the NIR-labeled BT is effective as a general marker of skeletal features and an indicator of the bone mineralization and remodeling processes. PMID- 21645492 TI - Crystal structure of UDP-galactose 4-epimerase from the hyperthermophilic archaeon Pyrobaculum calidifontis. AB - The crystal structure of a highly thermostable UDP-galactose 4-epimerase (GalE) from the hyperthermophilic archaeon Pyrobaculum calidifontis was determined at a resolution of 1.8A. The asymmetric unit contained one subunit, and the functional dimer was generated by a crystallographic two-fold axis. Each monomer consisted of a Rossmann-fold domain with NAD bound and a carboxyl terminal domain. The overall structure of P. calidifontis GalE showed significant similarity to the structures of the GalEs from Escherichia coli, human and Trypanosoma brucei. However, the sizes of several surface loops were markedly smaller in P. calidifontis GalE than the corresponding loops in the other enzymes. Structural comparison revealed that the presence of an extensive hydrophobic interaction at the subunit interface is likely the main factor contributing to the hyperthermostability of the P. calidifontis enzyme. Within the NAD-binding site of P. calidifontis GalE, a loop (NAD-binding loop) tightly holds the adenine ribose moiety of NAD. Moreover, a deletion mutant lacking this loop bound NAD in a loose, reversible manner. Thus the presence of the NAD-binding loop in GalE is largely responsible for preventing the release of the cofactor from the holoenzyme. PMID- 21645493 TI - The roles of chloroplast proteases in the biogenesis and maintenance of photosystem II. AB - Photosystem II (PSII) catalyzes one of the key reactions of photosynthesis, the light-driven conversion of water into oxygen. Although the structure and function of PSII have been well documented, our understanding of the biogenesis and maintenance of PSII protein complexes is still limited. A considerable number of auxiliary and regulatory proteins have been identified to be involved in the regulation of this process. The carboxy-terminal processing protease CtpA, the serine-type protease DegP and the ATP-dependent thylakoid-bound metalloprotease FtsH are critical for the biogenesis and maintenance of PSII. Here, we summarize and discuss the structural and functional aspects of these chloroplast proteases in these processes. This article is part of a Special Issue entitled: SI: Photosystem II. PMID- 21645494 TI - A minimal isoform of the TMEM16A protein associated with chloride channel activity. AB - TMEM16A protein, also known as anoctamin-1, has been recently identified as an essential component of Ca(2+)-activated Cl(-) channels. We previously reported the existence of different TMEM16A isoforms generated by alternative splicing. In the present study, we have determined the functional properties of a minimal TMEM16A protein. This isoform, called TMEM16A(0), has a significantly shortened amino-terminus and lacks three alternative segments localized in the intracellular regions of the protein (total length: 840 amino acids). TMEM16A(0) expression is associated with Ca(2+)-activated Cl(-) channel activity as measured by three different functional assays based on the halide-sensitive yellow fluorescent protein, short-circuit current recordings, and patch-clamp technique. However, compared to a longer isoform, TMEM16(abc) (total length: 982 amino acids), TMEM16A(0) completely lacks voltage-dependent activation. Furthermore, TMEM16A(0) and TMEM16A(abc) have similar but not identical responses to extracellular anion replacement, thus suggesting a difference in ion selectivity and conductance. Our results indicate that TMEM16A(0) has the basic domains required for anion transport and Ca(2+)-sensitivity. However, the absence of alternative segments, which are present in more complex isoforms of TMEM16A, modifies the channel gating and ion transport ability. PMID- 21645495 TI - Regulation of tumor suppressor gene FUS1 expression by the untranslated regions of mRNA in human lung cancer cells. AB - FUS1, also known as tumor suppressor candidate 2 (TUSC2), is a tumor suppressor gene located in the human chromosome 3p21.3 region. FUS1 mRNA transcripts could be detected on Northern blots in both normal lung and some lung cancer cell lines, but no endogenous FUS1 protein could be detected in a majority of lung cancer cell lines and small cell and non-small cell lung tumor tissues. However, mechanisms regulating FUS1 protein expression and its inactivation in primary lung cancer cells are largely unknown. In this study, we investigated the role of the 5'- and 3'-untranslated regions (UTRs) of the FUS1 gene transcript in the regulation of FUS1 protein expression. We identified RNA sequence elements in FUS1 UTRs that regulate FUS1 protein expression. We found that two small upstream open-reading frames in the 5'UTR of FUS1 mRNA could inhibit the translational initiation of FUS1 protein by interfering with the "scanning" of the ribosome initiation complexes. Several secondary RNA structural elements/motifs on the 3'UTR of FUS1 also exhibited a significant inhibitory effect on FUS1 protein expression. The 3'UTR-mediated regulatory effect on FUS1 protein expression was also differentially detected in normal lung epithelial and fibroblast cells compared with lung cancer cells. Our results provide new insight into the molecular mechanisms involved in the regulation of FUS1 expression. PMID- 21645496 TI - Killing of tumor cells: a drama in two acts. AB - Cancer still represents a major health problem worldwide, which urges the development of more effective strategies. Resistance to chemotherapy, a major obstacle for cancer eradication, is mainly related to an intrinsic failure to activate the apoptotic pathways. However, a protective effect of autophagy toward cancer cells has been recently observed, thus adding further complexity to the development of an effective approach counteracting cancer cell growth and improving the response to therapy. PMID- 21645497 TI - Differential dephosphorylation of the protein kinase C-zeta (PKCzeta) in an integrin alphaIIbbeta3-dependent manner in platelets. AB - Protein kinase C-zeta (PKCzeta), an atypical isoform of the PKC family of protein serine/threonine kinases, is expressed in human platelets. However, the mechanisms of its activation and the regulation of its activity in platelets are not known. We have found that under basal resting conditions, PKCzeta has a high phosphorylation status at the activation loop threonine 410 (T410) and the turn motif (autophosphorylation site) threonine 560 (T560), both of which have been shown to be important for its catalytic activity. After stimulation with agonist under stirring conditions, the T410 residue was dephosphorylated in a time- and concentration-dependent manner, while the T560 phosphorylation remained unaffected. The T410 dephosphorylation could be significantly prevented by blocking the binding of fibrinogen to integrin alphaIIbbeta3 with an antagonist, SC-57101; or by okadaic acid used at concentrations that inhibits protein serine/threonine phosphatases PP1 and PP2A in vitro. The dephosphorylation of T410 residue on PKCzeta was also observed in PP1cgamma null murine platelets after agonist stimulation, suggesting that other isoforms of PP1c or another phosphatase could be responsible for this dephosphorylation event. We conclude that human platelets express PKCzeta, and it may be constitutively phosphorylated at the activation loop threonine 410 and the turn motif threonine 560 under basal resting conditions, which are differentially dephosphorylated by outside-in signaling. This differential dephosphorylation of PKCzeta might be an important regulatory mechanism for platelet functional responses. PMID- 21645498 TI - Fragmentation of mitochondrial cardiolipin by copper ions in the Atp7b-/- mouse model of Wilson's disease. AB - Cellular copper overload as found in Wilson's disease may disturb mitochondrial function and integrity. Atp7b(-/-) mice accumulate copper in the liver and serve as an animal model for this inherited disease. The molecular mechanism of copper toxicity in hepatocytes is poorly understood. Total mitochondrial lipids from liver of wild-type mice were subjected to oxidative stress by the Cu(2+)/H(2)O(2)/ascorbate system. Phosphatidic acid (PA) and phosphatidylhydroxyacetone (PHA) were detected as cardiolipin fragmentation products by thin-layer chromatography combined with MALDI-TOF mass spectrometry in oxidized samples, but not in unperturbed ones. The formation of PA and PHA in copper-treated model membrane correlated well with the decrease of cardiolipin. Mitochondrial lipids from Atp7b(-/-) mice of different age were analyzed for the presence of PA. While 32-weeks old wild-type (control) and Atp7b(-/-) mice did not show any PA, there was a steady increase in the amount of this lipid in Atp7b(-/-) mice in contrast to control with increasing age. Hepatocytes from elder Atp7b(-/-)mice contained morphologically changed mitochondria unlike cells from wild-type animals of the same age. We concluded that free-radical fragmentation of cardiolipin with the formation of PA is a likely mechanism that damages mitochondria under conditions of oxidative stress due to copper overload. Our findings are relevant for better understanding of molecular mechanisms for liver damage found in Wilson's disease. PMID- 21645499 TI - Soluble vascular adhesion protein-1: decreased activity in the plasma of trauma victims and predictive marker for severity of traumatic brain injury. AB - BACKGROUND: This study done was to investigate the clinical significance of soluble vascular adhesion protein-1 (sVAP-1) activity in trauma patients with different patterns. METHODS: 96 patients with consecutive trauma >=15 years who were admitted to emergency department of the Second Affiliated Hospital, Shantou University Medical College, China, between January 2007 and December 2009 were enrolled in this study. Plasma was collected at admission. Injury-severity score (ISS) and Glasgow Coma Scale (GCS) were used to determine the patient conditions. sVAP-1 activity was determined by using the high performance liquid chromatographic (HPLC) system. RESULTS: Mean sVAP-1 activity in trauma patients was significantly lower than that of controls (P<0.0001), and the level was negatively correlated with circulating leucocytes and neutrophils (P<0.0001). There was a significant correlation between lower sVAP-1 activity and injury patterns. However, plasma sVAP-1 activity increased significantly in accordance with the severity of traumatic brain injury (TBI), and the patients with sVAP-1 value above 8.61 nmol/ml/h have much higher mortality rate (25.0%) than patients with sVAP-1 value lower than 8.61 nmol/ml/h (0.0%) (P=0.011). CONCLUSIONS: Trauma patients had a decreased sVAP-1 activity. However, isolated TBI patients with higher activity of sVAP-1 at admission were more likely to have a poor outcome. PMID- 21645500 TI - LecT-Hepa: A triplex lectin-antibody sandwich immunoassay for estimating the progression dynamics of liver fibrosis assisted by a bedside clinical chemistry analyzer and an automated pretreatment machine. AB - BACKGROUND: A quantitative analysis of glyco-alteration in serum glycoproteins provides glyco-parameters for estimating the progression of liver fibrosis. In the analysis of glycans, a manual pretreatment process for clinical specimens leads to a complicated manipulation and loss-of-clinical implementation of the assay. METHOD: We evaluated an automated triplex lectin-antibody sandwich immunoassay assisted by an automated protein purification system (ED-01) and a bedside clinical chemistry analyzer (HISCL) for the acquisition of two glyco parameters (AOL/DSA and MAL/DSA) derived from a fibrosis-related glyco-alteration of serum alpha1-acid glycoprotein (AGP). RESULTS: We adjusted the auto-machines with their accuracy set to CV <5.0% (ED-01) and <1.0% (HISCL). AGP samples were enriched from 275 serum specimens. Two glyco-parameters obtained by HISCL showed a linear correlation with that from a reported assay (R>0.90). The formula for monitoring fibrosis (LecT-Hepa) was given by a combination of the glyco parameters. This correlated with the fibrosis stage from biopsy (R=0.68) and diagnosed severe fibrosis and cirrhosis. It was superior to that of FIB-4 index. CONCLUSIONS: We automated a multilectin-assisted immunoassay with an order of magnitude reduction of operation time without any loss-of-accuracy. LecT-Hepa is a reliable method to assess fibrosis-dynamics from moderate fibrosis to cirrhosis. PMID- 21645501 TI - Dopamine improves hypothermic machine preservation of the liver. AB - BACKGROUND: Hypothermic machine preservation (HMP) is currently reconsidered as alternative to standard cold storage of organs from non-heart-beating donors. The present study was aimed at investigating the possible synergistic effect of HMP and the addition of dopamine to the circulating perfusate during preservation. METHODS: Cardiac arrest was induced in male Wistar rats (250-300 g) by phrenotomy. Thirty minutes later livers were flushed via the portal vein and subjected to 20 h of HMP at 5ml/min at 4 degrees C. During HMP the preservation solution was equilibrated with 100% oxygen and dopamine was added at 0, 10, 50 or 100 MUM (D0, D10, D50, D100; n=6 resp.). Graft viability was assessed thereafter upon warm reperfusion in vitro for 2h. RESULTS: During HMP, D50 and D100 significantly reduced hepatic release of ALT to about 50%. No influence of dopamine was found on vascular resistance, oxygen uptake or lactate production at any concentration. D50 significantly reduced enzyme release during reperfusion (~50%), enhanced bile flow and oxygen consumption. D10 was less effective while D100 even rose enzyme release compared with D0. Enhanced oxygen free radical mediated lipid peroxidation (LPO), found in the tissue of D0 livers was significantly reduced by D50; D50 significantly abrogated molecular upregulation of vWillebrand factor upon reperfusion suggesting vascular protection of the endothelial cell. CONCLUSION: Efficiency of HMP might be increased by stimulating livers with dopamine during ex vivo preservation, limiting vascular side effects and improving functional recovery upon early reperfusion. PMID- 21645502 TI - DNA methylation plays an important role in promoter choice and protein production at the mouse Dnmt3L locus. AB - The DNA methyltransferase 3-like (Dnmt3L) protein is a crucial cofactor in the germ line for the de novo methyltransferase Dnmt3a, which establishes imprints and represses transposable elements. We have previously shown that Dnmt3L transcription is regulated via three different promoters in mice, producing transcripts we term Dnmt3L(s) (stem cell), Dnmt3L(o) (oocyte) and Dnmt3L(at) (adult testis). Here we show that both Dnmt3L(s) and Dnmt3L(o) produce full length proteins but that the Dnmt3L(at) transcripts are not translated. Although not a canonical CpG island, the Dnmt3L(s) promoter is silenced by methylation during somatic differentiation in parallel with germ-cell-specific genes. During oocyte growth, Dnmt3L(s) also becomes heavily methylated and silenced and this requires its own gene product, since there is complete loss of methylation and derepression of transcription from this promoter in oocytes derived from Dnmt3L( /-) mice. Methylation of the Dnmt3L(s) promoter is established prior to the completion of imprinting and explains the requirement in mouse oocytes for the Dnmt3L(o) promoter, located in an intron of the neighboring unmethylated Aire gene. Overall these results give insight into how and why promoter switching at the mouse Dnmt3L locus occurs and provide one of the first examples of a non imprinted locus where methylation plays a role in promoter choice. The derepression of the Dnmt3L(s) promoter in the knockout oocytes also suggests that other non-imprinted loci may be dysregulated in these cells and contribute to the phenotype of the resultant mice. PMID- 21645503 TI - Role of hematopoietic prostaglandin D synthase in biphasic nasal obstruction in guinea pig model of experimental allergic rhinitis. AB - We investigated the role of hematopoietic prostaglandin D synthase (H-PGDS) in biphasic nasal obstruction in allergic rhinitis using a new specific inhibitor, (N-methoxy-N-methyl)-4-(5-benzoylbenzimidazole-2-yl)-3,5-dimethylpyrrole-2 carboxamide hydrochloride (TAS-204). First, we developed a novel guinea pig model of allergic rhinitis. Guinea pigs sensitized to ovalbumin without adjuvant were challenged with intranasal exposure to ovalbumin once a week. After the 3rd antigen challenge, they exhibited biphasic nasal obstruction. Additionally, analysis of nasal lavage fluid revealed an increase in the level of prostaglandin D(2) in both early and late phases. Treatment with oral TAS-204 for 15 days during the period of antigen challenges suppressed increases in nasal airway resistance in both phases. It is noteworthy that the late phase nasal obstruction was almost completely abrogated by inhibiting H-PGDS alone. Eosinophil infiltration in nasal lavage fluid and nasal hyperresponsiveness to histamine was also reduced by TAS-204 administration. These findings suggest that H-PGDS plays a critical role in the development of allergic rhinitis, especially in the induction of late phase nasal obstruction. PMID- 21645504 TI - Nicorandil attenuates cyclic strain-induced endothelin-1 expression via the induction of activating transcription factor 3 in human umbilical vein endothelial cells. AB - Nicorandil is an adenosine triphosphate-sensitive potassium channel opener that combines an organic nitrate and a nicotinamide group which respectively confer to nicorandil the additional properties of being a nitric oxide (NO) donor and antioxidant; it also induces vasodilation, decreases the blood pressure, and protects the heart. However, the intracellular mechanism of nicorandil remains to be delineated. The aims of this study were to test the hypothesis that nicorandil alters strain-induced endothelin-1 secretion and NO production, and to identify the putative underlying signaling pathways in human umbilical vein endothelial cells (HUVECs). Cultured HUVECs were exposed to cyclic strain in the presence of nicorandil; endothelin-1 expression was examined by reverse-transcriptase polymerase chain reaction and enzyme-linked immunosorbent assay. Activation of extracellular signal-regulated protein kinase (ERK), endothelial NO synthase (eNOS), and activating transcription factor (ATF)-3 was assessed by Western blot analysis. We show that nicorandil inhibited strain-induced endothelin-1 expression. Nicorandil also inhibited strain-increased reactive oxygen species formation and ERK phosphorylation. On the contrary, NO production, eNOS phosphorylation, and ATF3 expression were enhanced by nicorandil; however, L-NAME (an inhibitor of eNOS) and LY294002 (an inhibitor of phosphatidylinositol 3 kinase) inhibited nicorandil-increased ATF3 expression. Moreover, treatment of HUVECs with either an NO donor (NOC18; 3,3-bis[aminoethyl]-1-hydroxy-2-oxo-1 triazene) or an ATF3 activator (MG-132; carbobenzoxy-L-leucyl-L-leucyl-L leucinal) resulted in repression of strain-induced endothelin-1 expression. Furthermore, L-NAME, and small interfering RNA transfection of eNOS also partially attenuated the inhibitory effect of nicorandil on strain-induced endothelin-1 expression. We demonstrate for the first time that nicorandil inhibits strain-induced endothelin-1 secretion via an increase in NO and upregulation of ATF3 in HUVECs. This study provides important new insights into the molecular pathways that may contribute to the beneficial effects of nicorandil in the cardiovascular system. PMID- 21645505 TI - Ibuprofen hepatic encephalopathy, hepatomegaly, gastric lesion and gastric pentadecapeptide BPC 157 in rats. AB - Chronic ibuprofen (0.4 g/kg intraperitoneally, once daily for 4 weeks) evidenced a series of pathologies, not previously reported in ibuprofen-dosed rats, namely hepatic encephalopathy, gastric lesions, hepatomegaly, increased AST and ALT serum values with prolonged sedation/unconsciousness, and weight loss. In particular, ibuprofen toxicity was brain edema, particularly in the cerebellum, with the white matter being more affected than in gray matter. In addition, damaged and red neurons, in the absence of anti-inflammatory reaction was observed, particularly in the cerebral cortex and cerebellar nuclei, but was also present although to a lesser extent in the hippocampus, dentate nucleus and Purkinje cells. An anti-ulcer peptide shown to have no toxicity, the stable gastric pentadecapeptide BPC 157 (GEPPPGKPADDAGLV, MW 1419, 10 MUg, 10 ng/kg) inhibited the pathology seen with ibuprofen (i) when given intraperitoneally, immediately after ibuprofen daily or (ii) when given in drinking water (0.16 MUg, 0.16 ng/ml). Counteracted were all adverse effects, such as hepatic encephalopathy, the gastric lesions, hepatomegaly, increased liver serum values. In addition, BPC 157 treated rats showed no behavioral disturbances and maintained normal weight gain. Thus, apart from efficacy in inflammatory bowel disease and various wound treatments, BPC 157 was also effective when given after ibuprofen. PMID- 21645506 TI - A study on the mechanisms involving the anti-inflammatory effect of amitriptyline in carrageenan-induced paw edema in rats. AB - Anti-inflammatory effects of antidepressants have been reported in some studies, but the mechanisms underlying these effects remain unknown. Amitriptyline, a tricyclic antidepressant, is widely used in the management of psychological disorders and various types of pain, including neuropathic pain or fibromyalgia. In our previous work, we found the role of supraspinal mechanisms in the anti inflammatory effect of amitriptyline. In the line of the indicated study, we sought to evaluate the effects of intraperitoneal (i.p.) and intracerebroventricular (i.c.v.) application of amitriptyline in the carrageenan induced paw edema in rats in more details. Our findings confirmed that i.p. (40 and 80 mg/kg) and i.c.v. (100 MUg/rat) injection of amitriptyline inhibited carrageenan-induced inflammation at different times. We also found that both i.p. and i.c.v. amitriptyline significantly decreased migration of polymorphonuclear (PMN) leucocytes into the site of inflammation, according to pathological evidence and the activity of myeloperoxidase (MPO). Furthermore, i.p. amitriptyline at the applied doses markedly reduced interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha levels in the paw treated with carrageenan. Our results also showed that i.c.v. amitriptyline noticeably decreased the concentration of IL-1beta in the inflamed paws. The TNF-alpha levels reduced in the i.c.v. group, even though these reductions were not statistically significant. These results confirmed the anti-inflammatory effects of systemic and central amitriptyline in the carrageenan-induced paw edema in rats, and demonstrated that these effects mediated mostly through the inhibition of PMN cells migration and release of IL-1beta and TNF-alpha into the site of inflammation. PMID- 21645507 TI - Exenatide or glimepiride added to metformin on metabolic control and on insulin resistance in type 2 diabetic patients. AB - The aim of this study was to evaluate the effect of exenatide compared to glimepiride on body weight, glycemic control and insulin resistance in type 2 diabetic patients taking metformin. One hundred and eleven patients with uncontrolled type 2 diabetes mellitus and intolerant to metformin at the highest dosages (2500-3000 mg/day) were enrolled in this study. Patients were randomized to receive exenatide 5 MUg twice a day or glimepiride 1mg three times a day and titrated after 1 month to exenatide 10 MUg twice a day or glimepiride 2mg three times a day for 12 months in a randomized, single-blind, controlled study. We evaluated at the baseline and after 3, 6, 9, and 12 months these parameters: body weight, body mass index (BMI), HbA(1c), glycemic control, fasting plasma insulin, homeostasis model assessment insulin resistance index (HOMA-IR) index, adiponectin, tumor necrosis factor-alpha, and high sensitivity-C reactive protein. Both treatments gave a similar improvement of glycemic control, without any differences between the two groups. Only exenatide gave a decrease of BMI, insulin resistance parameters such as fasting plasma insulin, HOMA-IR, and adiponectin and a decrease of inflammatory parameters such as tumor necrosis factor-alpha, and high sensitivity-C reactive protein. Furthermore, the values obtained with exenatide were significantly better than the values recorded with glimepiride. We can conclude that exenatide was better than glimepiride in improving insulin resistance and inflammatory state. Furthermore, adiponectin increase, and tumor necrosis factor-alpha reduction seem to be related to weight loss obtained with exenatide. PMID- 21645508 TI - Role of tachykinin receptors in the modulation of colonic peristaltic activity in mice. AB - Tachykinins are important mediators of neuroneuronal and neuromuscular transmission in the gastrointestinal tract, however their contribution to colonic peristalsis in mice remains unclear. Therefore, our aim was to characterise the functional role of tachykinins in mediating peristalsis by evaluating the effect of selective tachykinin NK(1), NK(2) and NK(3) receptor agonists and antagonists on in vitro colonic peristaltic activity in mice. Using a modified Trendelenburg set-up, gradual distension of proximal and distal colonic segments evoked rhythmic, aborally migrating contractions. Peristaltic activity was assessed by quantifying the amplitude and interval of the corresponding pressure waves. Stimulation of NK(1) receptors showed regional differences as both the pressure amplitude and interval were enhanced in the distal colon without affecting peristalsis proximally. Blockade of NK(1) receptors reduced the peristaltic pressure amplitude in the proximal and distal colon while the interval was not significantly altered. NK(2) receptor stimulation resulted in a modest enhancement of the amplitude in proximal and distal segments and a slightly prolonged interval distally. Blockade of NK(2) receptors reduced the peristaltic pressure amplitude and interval in the distal colon. NK(3) receptor stimulation significantly augmented the amplitude in both segments and prolonged the interval distally. However, NK(3) receptor blockade had no effect on peristaltic activity. In conclusion, tachykinins contribute to colonic peristalsis in mice by acting mainly on NK(1) and NK(2) receptors and their effects show a proximal-to-distal gradient. NK(3) receptors might play a role in conditions of excess tachykinin release but appear not to be involved under the conditions of the present study. PMID- 21645509 TI - The effects of gabapentin in two animal models of co-morbid anxiety and visceral hypersensitivity. AB - Visceral hypersensitivity and an increased response to stress are two of the main symptoms of irritable bowel syndrome. Thus efforts to develop animal models of irritable bowel syndrome have centred on both of these parameters. The anticonvulsant gabapentin, which is widely used as an analgesic agent, also reduces anxiety. No data exists to our knowledge of the effects of gabapentin in animal models of co-morbid exaggerated stress response and visceral pain. Our aim was to assess the effect of gabapentin on stress and visceral hypersensitivity in two different animal models of irritable bowel syndrome. The animal models employed were the genetically susceptible Wistar Kyoto rat and the neonatally stressed maternal separation model. These animals were subjected to the open field paradigm to assess stress-induced defecation rates and colorectal distension to assess the level of visceral sensitivity. Gabapentin (30 mg/kg) prevented the stress-induced increase in faecal pellet output in the maternally separated rat, but not the Wistar Kyoto animals. On the other hand gabapentin (30 mg/kg) reduced the number of pain behaviours in response to colorectal distension in both models. These results show that whilst both models have similar responses to gabapentin in terms of visceral pain they differ in terms of their physiological response to stress. This indicates that the origin of anxiety and perhaps then visceral hypersensitivity differs in these models. Overall, these data suggest that gabapentin may be a useful treatment in disorders of co-morbid pain and an overactive stress system such as irritable bowel syndrome. PMID- 21645510 TI - Conditioned place preference studies with atomoxetine in an animal model of ADHD: effects of previous atomoxetine treatment. AB - To investigate the putative rewarding effects of atomoxetine, a non-stimulant medication for Attention-deficit/hyperactivity disorder (ADHD), we conducted conditioned place preference (CPP) tests in an animal model of ADHD, the spontaneously hypertensive rat (SHR). The effects of drug pre-exposure were also evaluated, thus, parallel experiments were done in rats which have undergone 14 days of atomoxetine treatment. The responses of SHR were compared with the rat strain representing the "normal" heterogeneous population, the Wistar rats. Neither rat strain showed significant CPP to atomoxetine. However, previous atomoxetine treatment produced place preference responses in rats, more profoundly in Wistar rats conditioned with the low and moderate atomoxetine doses. In conclusion, acute exposure to atomoxetine does not have any rewarding effect, however, drug pretreatment produces responses characteristic of reward or psychological dependence, more specifically in the "normal" vs. the ADHD animal model. The present findings call for more studies with atomoxetine, especially those that investigate the effects of long-term or chronic drug treatment. PMID- 21645511 TI - Biochemical characterization and in vitro activity of AZ513, a noncovalent, reversible, and noncompetitive inhibitor of fatty acid amide hydrolase. AB - Fatty acid amide hydrolase (FAAH) hydrolyzes several bioactive lipids including the endocannabinoid anandamide. Synthetic FAAH inhibitors are being generated to help define the biological role(s) of this enzyme, the lipids it degrades in vivo, and the disease states that might benefit from its pharmacological modulation. AZ513 inhibits human FAAH (IC(50)=551 nM), is 20-fold more potent against rat FAAH (IC(50)=27 nM), and is inactive at 10 MUM against the serine hydrolases acetylcholinesterase, thrombin, and trypsin. In contrast to most other potent FAAH inhibitors, AZ513 showed no evidence of covalently modifying the enzyme and displayed reversible inhibition. In an enzyme cross-competition assay, AZ513 did not compete with OL-135, an inhibitor that binds to the catalytic site in FAAH, which indicates that AZ513 does not bind to the catalytic site and is therefore noncompetitive with respect to substrate. AZ513 has good cell penetration as demonstrated by inhibition of anandamide hydrolysis in human FAAH transfected HEK293 cells (IC(50)=360 nM). AZ513 was tested in a rat spinal cord slice preparation where CB(1) activation reduces excitatory post-synaptic currents (EPSCs). In this native tissue assay of synaptic activity, AZ513 reduced EPSCs, which is consistent with inhibiting endogenous FAAH and augmenting endocannabinoid tone. AZ513 has a unique biochemical profile compared with other published FAAH inhibitors and will be a useful tool compound to further explore the role of FAAH in various biological processes. PMID- 21645512 TI - Therapeutic time window for treatment of focal cerebral ischemia reperfusion injury with XQ-1h in rats. AB - Pervious experimental studies have shown that XQ-1h has beneficial neuroprotective effect in the cerebral ischemia reperfusion injury. However, the therapeutic time window for treatment of focal cerebral ischemia reperfusion injury with XQ-1h is not clear. Under chloral hydrate anesthesia, transient focal cerebral ischemia was induced in rats by 2h of middle cerebral artery occlusion (MCAO), followed by 24h of reperfusion. Saline as vehicle or XQ-1h at the doses of 31.2, 15.6 and 7.8 mg/kg i.v. was administered at 0.5, 1, 2, 3h after induction of ischemia. Subsequently, 24h after MCAO brain edema, infarct volume, neurological deficits and cerebral blood flow were evaluated. Administrations of XQ-1h at the doses of 31.2mg/kg at 0.5, 1, and 2h after reperfusion of MCAO significantly reduced infarct rate (%) by 75.6% (5.2 +/- 1.7), 66.2% (7.2 +/- 1.9), and 47.9% (11.1 +/- 1.2), respectively. XQ-1h (31.2mg/kg) treatment, 0.5, 1, and 2h after reperfusion produced significant improvement in neurological score compared to vehicle-treated group (P<0.01). Administrations of XQ-1h at the doses of 31.2mg/kg and 15.6 mg/kg at 0.5, 1, and 2h after reperfusion of MCAO significantly increased cerebral blood flow (mv) by 16.9 +/- 1.9, 11.7 +/- 1.3, 9.5 +/- 1.0, respectively (P<0.01). In conclusion the therapeutic time window of XQ-1h for cerebral ischemia reperfusion injury is within 2h. Interestingly, we also discovered that the therapeutic time window of XQ-1h is deeply related with the activity of scavenging oxidative stress products. Further studies need to be conducted more drug combination therapy programs in order to assess the potential clinical application of XQ-1h. PMID- 21645513 TI - Effects of diphyllin as a novel V-ATPase inhibitor on gastric adenocarcinoma. AB - The natural compound diphyllin, a cytostatic lignan isolated from Cleistanthus collinus, can dramatically inhibit the proliferation and induce the apoptosis of human gastric cancer cells, SGC7901. Our study found that diphyllin can inhibit the expression of V-ATPases in a dose-dependent manner, decrease the internal pH (pHi) and reverse the transmembrane pH gradient in SGC7901 cells. Changes of the pH gradient were positively correlated with diphyllin concentration. Further study found that diphyllin treatment caused a decrease in phospho-LRP6, but not in LRP6. beta-catenin in Wnt/beta-catenin signaling and its target genes, c-myc and cyclin-D1, were also decreased with the inhibition of V-ATPases. Therefore, diphyllin could be characterized as a new V-ATPase inhibitor in treating gastric cancer and inhibiting the phosphorylation of LRP6 in Wnt/beta-catenin signaling. PMID- 21645514 TI - Ameliorative effect of caffeic acid against inflammatory pain in rodents. AB - The present study was designed to evaluate the antinociceptive profile of caffeic acid in mice and rats. Caffeic acid (5-100 mg/kg, p.o.), in a dose dependent manner inhibited acetic acid-induced writhing and late phase of formalin-induced pain in mice, with an ED(50) of 22.38 and 10.92 mg/kg, respectively. However, caffeic acid was ineffective in the hot plate and tail flick tests. Analgesic activity was also examined in carrageenan and lipopolysaccharide (LPS)-induced mechanical hyperalgesia in rats, where locally induced myeloperoxidase (MPO), malondialdehyde (MDA) and nitrite levels in foot pad were estimated by colorimetric assay. Oral administration of caffeic acid (200mg/kg, p.o.) showed analgesic activity similar to nimesulide (4 mg/kg, p.o.) and inhibited MPO, MDA and nitrite generation in the inflamed paw. Histological examination revealed reduction in neutrophil infiltration and protection of tissue damage by caffeic acid. These results suggest that caffeic acid exhibits peripheral analgesic effect in mice and rats and could be further examined for the treatment of chronic painful episodes. PMID- 21645515 TI - Comparative analysis of the role of small G proteins in cell migration and cell death: cytoprotective and promigratory effects of RalA. AB - Small G protein superfamily consists of more than 150 members, and is classified into six families: the Ras, Rho, Rab, Arf, Ran, and RGK families. They regulate a wide variety of cell functions such as cell proliferation/differentiation, cytoskeletal reorganization, vesicle trafficking, nucleocytoplasmic transport and microtubule organization. The small G proteins have also been shown to regulate cell death/survival and cell shape. In this study, we compared the role of representative members of the six families of small G proteins in cell migration and cell death/survival, two cellular phenotypes that are associated with inflammation, tumorigenesis, and metastasis. Our results show that small G proteins of the six families differentially regulate cell death and cell cycle distribution. In particular, our results indicate that Rho family of small G proteins is antiapoptotic. Ras, Rho, and Ran families promoted cell migration. There was no significant correlation between the cell death- and cell migration regulating activities of the small G proteins. Nevertheless, RalA was not only cytoprotective against multiple chemotherapeutic drugs, but also promigratory inducing stress fiber formation, which was accompanied by the activation of Akt and Erk pathways. Our study provides a framework for further systematic investigation of small G proteins in the perspectives of cell death/survival and motility in inflammation and cancer. PMID- 21645516 TI - Sociosexuality moderates the association between testosterone and relationship status in men and women. AB - Single individuals typically have higher testosterone compared to those who are partnered, suggesting that individual differences in testosterone are associated with mating effort, or people's motivation to find a sexual partner. However, there is less consistent evidence for links between testosterone and sociosexuality, or people's orientation toward uncommitted sexual activity. Based on Penke and Asendorpf's (2008) conceptualization, we propose that a more nuanced measure of sociosexuality may reveal more robust associations with testosterone. In the current study, we assessed relations between three components of sociosexuality--desire, behavior, and attitudes--and endogenous testosterone levels in men and women. We found that partnered status was indeed associated with lower testosterone in both men and women, but only among those who reported more restricted sociosexuality. Partnered men who reported greater desire for uncommitted sexual activity had testosterone levels that were comparable to those of single men; partnered women who reported more frequent uncommitted sexual behavior had testosterone levels that were comparable to those of single women. These findings provide new evidence that people's orientations toward sexual relationships, in combination with their relationship status, are associated with individual differences in testosterone. The current results are also among the first to demonstrate sociosexuality-testosterone associations in both men and women, and they reveal that the nature of these associations varies by gender. Together, these findings highlight the utility of a multifaceted conceptualization of sociosexuality and the implications of this conceptualization for neuroendocrine processes. PMID- 21645517 TI - Bi-modal dose-dependent cardiac response to tetrahydrobiopterin in pressure overload induced hypertrophy and heart failure. AB - The exogenous administration of tetrahydrobiopterin (BH4), an essential cofactor of nitric oxide synthase (NOS), has been shown to reduce left ventricular hypertrophy, fibrosis, and cardiac dysfunction in mice with pre-established heart disease induced by pressure-overload. In this setting, BH4 re-coupled endothelial NOS (eNOS), with subsequent reduction of NOS-dependent oxidative stress and reversal of maladaptive remodeling. However, recent studies suggest the effective BH4 dosing may be narrower than previously thought, potentially due to its oxidation upon oral consumption. Accordingly, we assessed the dose response of daily oral synthetic sapropterin dihydrochloride (6-R-l-erythro-5,6,7,8 tetrahydrobiopterin, 6R-BH4) on pre-established pressure-overload cardiac disease. Mice (n=64) were administered 0-400mg/kg/d BH4 by ingesting small pre made pellets (consumed over 15-30 min). In a dose range of 36-200mg/kg/d, 6R-BH4 suppressed cardiac chamber remodeling, hypertrophy, fibrosis, and oxidative stress with pressure-overload. However, at both lower and higher doses, BH4 had less or no ameliorative effects. The effective doses correlated with a higher myocardial BH4/BH2 ratio. However, BH2 rose linearly with dose, and at the 400mg/kg/d, this lowered the BH4/BH2 ratio back toward control. These results expose a potential limitation for the clinical use of BH4, as variability of cellular redox and perhaps heart disease could produce a variable therapeutic window among individuals. This article is part of a special issue entitled ''Key Signaling Molecules in Hypertrophy and Heart Failure.'' PMID- 21645518 TI - Dynamic modulation of Ca2+ sparks by mitochondrial oscillations in isolated guinea pig cardiomyocytes under oxidative stress. AB - Local control of Ca(2+)-induced Ca(2+) release (CICR) depends on the spatial organization of L-type Ca(2+) channels and ryanodine receptors (RyR) in the dyad. Analogously, Ca(2+) uptake by mitochondria is facilitated by their close proximity to the Ca(2+) release sites, a process required for stimulating oxidative phosphorylation during changes in work. Mitochondrial feedback on CICR is less well understood. Since mitochondria are a primary source of reactive oxygen species (ROS), they could potentially influence the cytosolic redox state, in turn altering RyR open probability. We have shown that self-sustained oscillations in mitochondrial inner membrane potential (DeltaPsi(m)), NADH, ROS, and reduced glutathione (GSH) can be triggered by a laser flash in cardiomyocytes. Here, we employ this method to directly examine how acute changes in energy state dynamically influence resting Ca(2+) spark occurrence and properties. Two-photon laser scanning microscopy was used to monitor cytosolic Ca(2+) (or ROS), DeltaPsi(m), and NADH (or GSH) simultaneously in isolated guinea pig cardiomyocytes. Resting Ca(2+) spark frequency increased with each DeltaPsi(m) depolarization and decreased with DeltaPsi(m) repolarization without affecting Ca(2+) spark amplitude or time-to-peak. Stabilization of mitochondrial energetics by pretreatment with the superoxide scavenger TMPyP, or by acute addition of 4'-chlorodiazepam, a mitochondrial benzodiazepine receptor antagonist that blocks the inner membrane anion channel, prevented or reversed, respectively, the increased spark frequency. Cyclosporine A did not block the DeltaPsi(m) oscillations or prevent Ca(2+) spark modulation by DeltaPsi(m). The results support the hypothesis that mitochondria exert an influential role on the redox environment of the Ca(2+) handling subsystem, with mechanistic implications for the pathophysiology of cardiac disease. PMID- 21645519 TI - Pharmacologic and genetic strategies to enhance cell therapy for cardiac regeneration. AB - Cell-based therapy is emerging as an exciting potential therapeutic approach for cardiac regeneration following myocardial infarction (MI). As heart failure (HF) prevalence increases over time, development of new interventions designed to aid cardiac recovery from injury are crucial and should be considered more broadly. In this regard, substantial efforts to enhance the efficacy and safety of cell therapy are continuously growing along several fronts, including modifications to improve the reprogramming efficiency of inducible pluripotent stem cells (iPS), genetic engineering of adult stem cells, and administration of growth factors or small molecules to activate regenerative pathways in the injured heart. These interventions are emerging as potential therapeutic alternatives and/or adjuncts based on their potential to promote stem cell homing, proliferation, differentiation, and/or survival. Given the promise of therapeutic interventions to enhance the regenerative capacity of multipotent stem cells as well as specifically guide endogenous or exogenous stem cells into a cardiac lineage, their application in cardiac regenerative medicine should be the focus of future clinical research. This article is part of a special issue entitled "Key Signaling Molecules in Hypertrophy and Heart Failure." PMID- 21645520 TI - Dynamics may significantly influence the estimation of interatomic distances in biomolecular X-ray structures. AB - Atomic positions obtained by X-ray crystallography are time and space averages over many molecules in the crystal. Importantly, interatomic distances, calculated between such average positions and frequently used in structural and mechanistic analyses, can be substantially different from the more appropriate time-average and ensemble-average interatomic distances. Using crystallographic B factors, one can deduce corrections, which have so far been applied exclusively to small molecules, to obtain correct average distances as a function of the type of atomic motion. Here, using 4774 high-quality protein X-ray structures, we study the significance of such corrections for different types of atomic motion. Importantly, we show that for distances shorter than 5 A, corrections greater than 0.5 A may apply, especially for noncorrelated or anticorrelated motion. For example, 14% of the studied structures have at least one pair of atoms with a correction of >=0.5 A in the case of noncorrelated motion. Using molecular dynamics simulations of villin headpiece, ubiquitin, and SH3 domain unit cells, we demonstrate that the majority of average interatomic distances in these proteins agree with noncorrelated corrections, suggesting that such deviations may be truly relevant. Importantly, we demonstrate that the corrections do not significantly affect stereochemistry and the overall quality of final refined X ray structures, but can provide marked improvements in starting unrefined models obtained from low-resolution X-ray data. Finally, we illustrate the potential mechanistic and biological significance of the calculated corrections for KcsA ion channel and show that they provide indirect evidence that motions in its selectivity filter are highly correlated. PMID- 21645521 TI - Amyloid fibrillation kinetics: insight from atomistic nucleation theory. AB - We consider the nucleation of nanosized amyloid fibrils composed of successively layered beta-sheets at the molecular level when this process takes place by direct polymerization of protein segments (beta-strands) into beta-sheets. Application of the atomistic nucleation theory (ANT) to amyloid nucleation of beta(2)-microglobulin and amyloid beta(40) allows us to predict the fibril nucleus size and the fibril nucleation rate as functions of the supersaturation of the protein solution. The ANT predictions are compared to recent time-resolved optical experiments where they measure the effect of the protein concentration and mutations on the initial lag time before amyloid fibrils form in the protein solution. The presented analysis reveals the general principles underlying the nucleation kinetics of nanosized amyloid fibrils and indicates that it can be treated in the framework of existing general theories of the nucleation of new phases. PMID- 21645522 TI - Structural and computational investigations of VIM-7: insights into the substrate specificity of vim metallo-beta-lactamases. AB - The presence of metallo-beta-lactamases (MBLs) in many clinically important human bacterial pathogens limits treatment options, as these enzymes efficiently hydrolyze nearly all beta-lactam antibiotics. VIM enzymes are among the most widely distributed MBLs, but many of the individual VIM subtypes remain poorly characterized. Pseudomonas aeruginosa VIM-7 is the most divergent among VIM-type MBLs in terms of amino acid sequence. Here we present crystal structures of VIM-7 as the native enzyme, with Cys221 oxidized (VIM-7-Ox), and with a sulfur atom bridging the two active-site zinc ions (VIM-7-S). Comparison with VIM-2 and VIM-4 structures suggests an explanation for the reduced catalytic efficiency of VIM-7 against cephalosporins with a positively charged cyclic substituent at the C3 position (e.g., ceftazidime). Kinetic variations are attributed to substitutions in residues 60-66 (that form a loop adjacent to the active site previously implicated in substrate binding) and to the disruption of two hydrogen-bonding clusters through substitutions at positions 218 and 224. Furthermore, the less negatively charged surface of VIM-7 (compared to VIM-2) may also contribute to the reduced hydrolytic efficiency. Docking of the cephalosporins ceftazidime and cefotaxime into the VIM-2 and VIM-7 structures reveals that amino acid substitutions may cause the mode of substrate binding to differ between the two enzymes. Our structures thus provide new insights into the variation in substrate specificity that is evident across this family of clinically important enzymes. PMID- 21645523 TI - Effect of the endothelial glycocalyx layer on arterial LDL transport under normal and high pressure. AB - To quantitatively investigate the role of the endothelial glycocalyx layer (EGL) in protecting the artery from excessive infiltration of atherogenic lipids such as low density lipoproteins (LDLs), a multilayer model with the EGL of an arterial segment was developed to numerically simulate the flow and the transport of LDLs under normal and high pressure. The transport parameters of the layers of the model were obtained from the hydrodynamic theory, the stochastic theory, and from the literature. The results showed that the increase in the thickness of the EGL could lead to a sharp drop in LDL accumulation in the intima. A partial damage to the EGL could compromise its barrier function, hence leading to enhanced infiltration/accumulation of LDLs within the wall of the arterial model. Without the EGL, hypertension could lead to a significantly enhanced LDL transport into the wall of the model. However, the intact EGL could protect the arterial wall from hypertension so that the LDL concentration in the intima layer was almost the same as that under normal pressure conditions. The results also showed that LDL concentration within the arterial wall increased with Phi (the fraction of leaky junctions) on the intima layer. The increase in LDL concentration with Phi was much more dramatic for the model without the EGL. For instance, without the EGL, a Phi of 0.0005 could lead LDL concentration within the arterial wall to be even higher than that predicted for the EGL intact model with a Phi of 0.002. In conclusion, an intact EGL with a sufficient thickness may act as a barrier to LDL infiltration into the arterial wall and has the potential to suppress the hypertension-driven hike of LDL infiltration/accumulation in the arterial wall. PMID- 21645524 TI - A method to calculate binding equilibrium concentrations in the allosteric ternary complex model that supports ligand depletion. AB - The allosteric ternary complex model is frequently used in pharmacology to represent the interaction of a receptor R with two ligands A and B. Certain well known formulas are routinely used to calculate the fractions of the receptor bound at equilibrium with A only, B only, and both A and B. However, it is often omitted that these classical formulas presume that there is no ligand depletion, i.e. that the equilibrium concentrations [A] and [B] of the ligands are well approximated by their total concentrations [A](T) and [B](T). We present a calculation method which is applicable without this or any restrictions. The equilibrium concentration [R] of the receptor is implicitly characterized by an equation which is solved with a very simple convergent numerical algorithm. The concentrations [A] and [B] are given by explicit formulas in terms of [R]. The required parameters are the equilibrium dissociation constants K(A) and K(B), the cooperativity factor alpha, and the total concentrations [R](T), [A](T) and [B](T). PMID- 21645525 TI - Airborne urban particles (Milan winter-PM2.5) cause mitotic arrest and cell death: Effects on DNA, mitochondria, AhR binding and spindle organization. AB - Airborne particulate matter (PM) is considered to be an important contributor to lung diseases. In the present study we report that Milan winter-PM2.5 inhibited proliferation in human bronchial epithelial cells (BEAS-2B) by inducing mitotic arrest. The cell cycle arrest was followed by an increase in mitotic-apoptotic cells, mitotic slippage and finally an increase in "classical" apoptotic cells. Exposure to winter-PM10 induced only a slight effect which may be due to the presence of PM2.5 in this fraction while pure combustion particles failed to disturb mitosis. Fewer cells expressing the mitosis marker phospho-histone H3 compared to cells with condensed chromosomes, suggest that PM2.5 induced premature mitosis. PM2.5 was internalized into the cells and often localized in laminar organelles, although particles without apparent plasma membrane covering were also seen. In PM-containing cells mitochondria and lysosomes were often damaged, and in mitotic cells fragmented chromosomes often appeared. PM2.5 induced DNA strands breaks and triggered a DNA-damage response characterized by increased phosphorylation of ATM, Chk2 and H2AX; as well as induced a marked increase in expression of the aryl hydrocarbon receptor (AhR)-regulated genes, CYP1A1, CYP1B1 and AhRR. Furthermore, some disturbance of the organization of microtubules was indicated. It is hypothesized that the induced mitotic arrest and following cell death was due to a premature chromosome condensation caused by a combination of DNA, mitochondrial and spindle damage. PMID- 21645526 TI - Radiation-induced bystander effect in healthy G(o) human lymphocytes: biological and clinical significance. AB - To study the bystander effects, G(0) human peripheral blood lymphocytes were X irradiated with 0.1, 0.5 and 3 Gy. After 24h, cell-free conditioned media from irradiated cultures were transferred to unexposed lymphocytes. Following 48 h of medium transfer, viability, induction of apoptosis, telomere shortening, reactive oxygen species (ROS) levels and micronuclei (after stimulation) were analyzed. A statistically significant decrement in cell viability, concomitant with the loss of mitochondrial membrane potential, telomere shortening, increases in hydrogen peroxide (H(2)O(2)) and superoxide anion (O(2)(-)) with depletion of intracellular glutathione (GSH) level, and higher frequencies of micronuclei, were observed in bystander lymphocytes incubated with medium from 0.5 and 3 Gy irradiated samples, compared to lymphocytes unexposed. Furthermore, no statistically significant difference between the response to 0.5 and 3 Gy of irradiation in bystander lymphocytes, was found. However, when lymphocytes were irradiated with 0.1 Gy, no bystander effect with regard to viability, apoptosis, telomere length, and micronuclei was observed, although a high production of ROS level persisted. Radiation in the presence of the radical scavenger dimethyl sulfoxide (DMSO) suppressed oxidative stress induced by 3 Gy of X-rays with the effective elimination of bystander effects, suggesting a correlation between ROS and bystander signal formation in irradiated cells. The data propose that bystander effect might be mostly due to the reactions of radiation induced free radicals on DNA, with the existence of a threshold at which the bystander signal is not operative (0.1 Gy dose of X-rays). Our results may have clinical implications for health risk associated with radiation exposure. PMID- 21645527 TI - Transient receptor potential vanilloid 1 agonists modulate hippocampal CA1 LTP via the GABAergic system. AB - Transient receptor potential vanilloid 1 (TRPV1) was shown to modulate hippocampal CA1 pyramidal cell synaptic plasticity, including long-term potentiation (LTP) and long-term depression (LTD). Synaptic plasticity is the cellular mechanism thought to mediate declarative learning and memory in the hippocampus. Although TRPV1 is involved in modulating hippocampal plasticity, it has yet to be determined how TRPV1 mediates its effects. Using field electrophysiology in hippocampal CA1 stratum radiatum we investigated how TRPV1 agonists modulate LTP, low frequency stimulation-induced LTD, and (RS)-3,5 dihydroxyphenylglycine (DHPG)-induced LTD. First we confirmed that TRPV1 agonists induce enhancement of CA1 pyramidal cell LTP in the absence the GABA(A) receptor antagonist picrotoxin. Because it was recently determined that TRPV1 mediates a novel form of LTD in CA1 inhibitory GABAergic interneurons, which can disinhibit CA1 pyramidal cells, we used picrotoxin to block the effect of the GABAergic circuitry on CA1 LTP. When using picrotoxin, the TRPV1 agonist-induced enhancement of CA1 LTP was eliminated suggesting that the GABAergic circuitry is required for TRPV1 agonist mediated increases. Regarding LTD, in contrast to previously reported data, we did not see TRPV1 agonist-mediated effect on low frequency-induced stimulus LTD. However, during DHPG-induced LTD, TRPV1 was involved in the acute, but not the long-term depression phase of this plasticity. In summary, our findings support TRPV1 agonist involvement in hippocampal synaptic plasticity, including its enhancement of CA1 LTP. We demonstrate that the enhancement mediated by TRPV1 agonists requires GABA input to pyramidal cells thus providing a mechanism for how TRPV1 agonists modulate hippocampal synaptic plasticity. PMID- 21645528 TI - Functional crosstalk and heteromerization of serotonin 5-HT2A and dopamine D2 receptors. AB - The serotonin 5-HT(2A) receptor (5-HT(2A)R) and dopamine D(2) receptor (D(2)R) are high-affinity G protein-coupled receptor targets for two different classes of antipsychotic drugs used to treat schizophrenia. Interestingly, the antipsychotic effects are not based on the regulation of same signaling mediators since activation of the 5-HT(2A)R and of the D(2)R regulate G(q/11) protein and G(i/o) protein, respectively. Here we use radioligand binding and second messenger production assays to provide evidence for a functional crosstalk between 5 HT(2A)R and D(2)R in brain and in HEK293 cells. D(2)R activation increases the hallucinogenic agonist affinity for 5-HT(2A)R and decreases the 5-HT(2A)R induced inositol phosphate production. In vivo, 5-HT(2A)R expression is necessary for the full effects of D(2)R antagonist on MK-801-induced locomotor activity. Co immunoprecipitation studies show that the two receptors can physically interact in HEK293 cells and raise the possibility that a receptor heterocomplex mediates the crosstalk observed. The existence of this 5-HT(2A)R-D(2)R heteromer and crosstalk may have implications for diseases involving alterations of serotonin and dopamine systems and for the development of new classes of therapeutic drugs. PMID- 21645529 TI - Acute morphine administration and withdrawal from chronic morphine increase afterdepolarization amplitude in rat supraoptic nucleus neurons in hypothalamic explants. AB - Supraoptic nucleus (SON) neurons secrete either oxytocin or vasopressin into the bloodstream from their axon terminals in the posterior pituitary gland. SON neurons are powerfully inhibited by the classical MU-opioid receptor agonist, morphine. Oxytocin neurons develop morphine dependence when chronically exposed to this opiate, and undergo robust withdrawal excitation when morphine is subsequently acutely antagonized by naloxone. Morphine withdrawal excitation is evident as an increased firing rate and is associated with an increased post spike excitability that is consistent with the expression of an enhanced post spike afterdepolarization (ADP) during withdrawal. Here, we used sharp electrode recording from SON neurons in hypothalamic explants from morphine naive and morphine treated rats to determine the effects of morphine on the ADP, and to test the hypothesis that morphine withdrawal increases ADP amplitude in SON neurons. Acute morphine administration (0.05-5.0 MUM) caused a dose-dependent hyperpolarization of SON neurons that was reversed by concomitant administration of 10 MUM naloxone, or by washout of morphine; counter-intuitively, acute exposure to 5 MUM morphine increased ADP amplitude by 78 +/- 11% (mean +/- SEM). Naloxone-precipitated morphine withdrawal did not alter baseline membrane potential in SON neurons from morphine treated rats, but increased ADP amplitude by 48 +/- 11%; this represents a hyper-activation of ADPs because the basal amplitude of the ADP was similar in SON neurons recorded from explants prepared from morphine naive and morphine treated rats. Hence, an enhanced ADP might contribute to morphine withdrawal excitation of oxytocin neurons. PMID- 21645530 TI - Chronic exposure to anabolic androgenic steroids alters activity and synaptic function in neuroendocrine control regions of the female mouse. AB - Disruption of reproductive function is a hallmark of abuse of anabolic androgenic steroids (AAS) in female subjects. To understand the central actions of AAS, patch clamp recordings were made in estrous, diestrous and AAS-treated mice from gonadotropin releasing hormone (GnRH) neurons, neurons in the medial preoptic area (mPOA) and neurons in the anteroventroperiventricular nucleus (AVPV); regions known to provide GABAergic and kisspeptin inputs to the GnRH cells. Action potential (AP) frequency was significantly higher in GnRH neurons of estrous mice than in AAS-treated or diestrous animals. No significant differences in AAS-treated, estrous or diestrous mice were evident in the amplitude or kinetics of spontaneous postsynaptic currents (sPSCs), miniature PSCs or tonic currents mediated by GABA(A) receptors or in GABA(A) receptor subunit expression in GnRH neurons. In contrast, the frequency of GABA(A) receptor-mediated sPSCs in GnRH neurons showed an inverse correlation with AP frequency across the three hormonal states. Surprisingly, AP activity in the medial preoptic area (mPOA), a likely source of GABAergic afferents to GnRH cells, did not vary in concert with the sPSCs in the GnRH neurons. Furthermore, pharmacological blockade of GABA(A) receptors did not alter the pattern in which there was lower AP frequency in GnRH neurons of AAS-treated and diestrous versus estrous mice. These data suggest that AAS do not impose their effects either directly on GnRH neurons or on putative GABAergic afferents in the mPOA. AP activity recorded from neurons in kisspeptin rich regions of the AVPV and the expression of kisspeptin mRNA and peptide did vary coordinately with AP activity in GnRH neurons. Our data demonstrate that AAS treatment imposes a "diestrous-like" pattern of activity in GnRH neurons and suggest that this effect may arise from suppression of presynaptic kisspeptin mediated excitatory drive arising from the AVPV. The actions of AAS on neuroendocrine regulatory circuits may contribute the disruption of reproductive function observed in steroid abuse. PMID- 21645531 TI - Cannabinoid receptor antagonists AM251 and AM630 activate TRPA1 in sensory neurons. AB - Cannabinoid receptor antagonists have been utilized extensively in vivo as well as in vitro, but their selectivity has not been fully examined. We investigated activation of sensory neurons by two cannabinoid antagonists - AM251 and AM630. AM251 and AM630 activated trigeminal (TG) sensory neurons in a concentration dependent fashion (threshold 1 MUM). AM251 and AM630 responses are mediated by the TRPA1 channel in a majority (90-95%) of small-to-medium TG sensory neurons. AM630 (1-100 MUM), but not AM251, was a significantly more potent agonist in cells co-expressing both TRPA1 and TRPV1 channels. We next evaluated AM630 and AM251 effects on TRPV1- and TRPA1-mediated responses in TG neurons. Capsaicin (CAP) effects were inhibited by pre-treatment with AM630, but not AM251. Mustard oil (MO) and WIN55,212-2 (WIN) TRPA1 mediated responses were also inhibited by pre-treatment with AM630, but not AM251 (25 uM each). Co-treatment of neurons with WIN and either AM630 or AM251 had opposite effects: AM630 sensitized WIN responses, whereas AM251 inhibited WIN responses. WIN-induced inhibition of CAP responses in sensory neurons was reversed by AM630 pre-treatment and AM251 co treatment (25 MUM each), as these conditions inhibit WIN responses. Hindpaw injections of AM630 and AM251 did not produce nocifensive behaviors. However, both compounds modulated CAP-induced thermal hyperalgesia in wild-type mice and rats, but not TRPA1 null-mutant mice. AMs also partially regulate WIN inhibition of CAP-induced thermal hyperalgesia in a TRPA1-dependent fashion. In summary, these findings demonstrate alternative targets for the cannabinoid antagonists, AM251 and AM630, in peripheral antihyperalgesia which involve certain TRP channels. PMID- 21645532 TI - Peripheral inflammation upregulates P2X receptor expression in satellite glial cells of mouse trigeminal ganglia: a calcium imaging study. AB - Satellite glial cells (SGCs) in sensory ganglia are altered structurally and biochemically as a result of nerve injury. Whereas there is ample evidence that P2 purinergic receptors in central glial cells are altered after injury, there is very little information on similar changes in SGCs, although it is well established that SGCs are endowed with P2 receptors. Using calcium imaging, we characterized changes in P2 receptors in SGCs from mouse trigeminal ganglia in short-term cultures. Seven days after the induction of submandibular inflammation with complete Freund's adjuvant, there was a marked increase in the sensitivity of SGCs to ATP, with the threshold of activation decreasing from 5 MUM to 10 nM. A similar observation was made in the intact trigeminal ganglion after infra orbital nerve axotomy. Using pharmacological tools, we investigated the receptor mechanisms underlying these changes in cultured SGCs. We found that in control tissues response to ATP was mediated by P2Y (metabotropic) receptors, whereas after inflammation the response was mediated predominantly by P2X (ionotropic) receptors. As the contribution of P2X1,3,6 receptors was excluded, and the sensitivity to a P2X7 agonist did not change after inflammation, it appears that after inflammation the responses to ATP are largely due to P2X2 and/or 5 receptors, with a possible contribution of P2X4 receptors. We conclude that inflammation induced a large increase in the sensitivity of SGCs to ATP, which involved a switch from P2Y to P2X receptors. We propose that the over 100-fold augmented sensitivity of SGCs to ATP after injury may contribute to chronic pain states. PMID- 21645533 TI - Effects of high frequency electrical stimulation and R-verapamil on seizure susceptibility and glutamate and GABA release in a model of phenytoin-resistant seizures. AB - The present study was focused to characterize the effects of intrahippocampal application of R-verapamil, a P-glycoprotein blocker, and High Frequency Electrical Stimulation (HFS) at 130 Hz, on seizure susceptibility and extracellular concentrations of glutamate and gamma-aminobutyric acid (GABA) in hippocampus of kindled rats with drug-resistant seizures. Fully kindled rats classified in responsive and non-responsive to phenytoin were used for this purpose. In contrast with responsive animals, non-responsive rats showed lower afterdischarge threshold (ADT) values in pre-kindling conditions and required less number of kindling trials to achieve the kindled state. Once the animals attained the kindled state, both epileptic groups presented high glutamate and low GABA interictal release, effect more evident in non-responsive rats. In hippocampus of responsive animals, GABA levels demonstrated two increases at 120 and 240 min after the ictal event, a situation no detected for non-responsive rats. Kindled animals receiving hippocampal HFS showed augmented ADT, an effect associated with enhanced GABA release in responsive rats. Intrahippocampal perfusion of R-verapamil (5 mM) decreased the seizure susceptibility (high ADT values), enhanced the interictal GABA release and the postictal levels of glutamate and GABA in responsive and non-responsive rats. It is conclude that alterations of glutamate and GABA release in the epileptic hippocampus of non responsive animals resemble those found in hippocampus of patients with refractory TLE. In addition, intrahippocampal application of HFS and R-verapamil modifies the amino acid release and reduces the seizure susceptibility of both, responsive and non-responsive rats. PMID- 21645534 TI - The functional 5-HT1A receptor polymorphism affects response inhibition processes in a context-dependent manner. AB - Cognitive control processes may depend on contextual information, sometimes improving performance, but impairing performance if expectancies about forthcoming events induce pre-potent responses. The neurobiological bases of these effects are not understood. Here, we examine context-dependent variations of response control processes using the AX-CPT task with respect to the relevance of the functional serotonin 1A receptor polymorphism (5-HT1A C(-1019)G) in a sample of healthy subjects (N=90) by means of event-related potentials (ERPs). The results show that, when context information is helpful to drive behavioural performance, carriers of the -1019G allele reveal compromised cognitive control. Yet, they show enhanced task performance when strong context representations would lead to declines in behavioural control. These findings are paralleled by modulations of the (Nogo)-P3 ERP-component. These results show for the first time that, even though the -1019G allele enhances the risk to develop anxiety disorders, it also confers an advantage to its carriers in terms of better cognitive control processes in conditions where contextual information compromises cognitive control. Effects of the 5-HT1A C(-1019)G polymorphism were further modulated by anxiety sensitivity. As the functional effect of the 5-HT1A C(-1019)G polymorphism has previously been shown to be rather specific for serotonergic 1A autoreceptors in the dorsal raphe nucleus (DRN), the results suggest that contextual modulations in cognitive control may be exerted by the DRN. PMID- 21645535 TI - Norrish-Prins reaction as a key step in the synthesis of 14beta-hydroxy-5alpha (or 5beta or Delta(5,6))-pregnane derivatives. AB - Numerous bioactive glycosteroids are characterized by aglycones bearing a 14beta hydroxy pregnane skeleton like boucerin and isoramanone. In general, the syntheses of the latter are achieved by acidic hydrolysis of the corresponding glycosteroids. These aglycones were also obtained by a combined Norrish type I Prins reaction starting from the corresponding 12-keto-pregnane derivatives. However, for the Norrish-Prins reaction, no reports describe the influence of the A/B ring junction (cis or trans or Delta(5,6) double bond) or the influence of the substitution pattern at position 20. Herein, we describe the use of Norrish type I-Prins reactions to synthesize isoramanone and boucerin derivatives and their A/B cis and trans analogs. The influence of the parameters mentioned above is also presented. These studies showed that the A/B ring junction has little influence on the Norrish type I-Prins reaction but that the substitution pattern at position 20 is important. The presence of a dioxolane group induced not only the formation of the desired 14beta-hydroxy pregnane derivatives in the highest yields but also the formation of new spiro derivatives. PMID- 21645536 TI - The physiology of integrated nuclear and extranuclear steroid signaling: introductory comments. PMID- 21645537 TI - Mitochondrial transfection for studying organellar DNA repair, genome maintenance and aging. AB - Maintenance of the mitochondrial genome is a major challenge for cells, particularly as they begin to age. Although it is established that organelles possess regular DNA repair pathways, many aspects of these complex processes and of their regulation remain to be investigated. Mitochondrial transfection of isolated organelles and in whole cells with customized DNA synthesized to contain defined lesions has wide prospects for deciphering repair mechanisms in a physiological context. We document here the strategies currently developed to transfer DNA of interest into mitochondria. Methodologies with isolated mitochondria claim to exploit the protein import pathway or the natural competence of the organelles, to permeate the membranes or to use conjugal transfer from bacteria. Besides biolistics, which remains restricted to yeast and Chlamydomonas reinhardtii, nanocarriers or fusion proteins have been explored as methods to target custom DNA into mitochondria in intact cells. In further approaches, whole mitochondria have been transferred into recipient cells. Repair failure or error-prone repair leads to mutations which potentially could be rescued by allotopic expression of proteins. The relevance of the different approaches for the analysis of mitochondrial DNA repair mechanisms and of aging is discussed. PMID- 21645539 TI - Worldwide prevalence of physical inactivity calls for worldwide actions. AB - OBJECTIVE: This commentary offers a discussion of the paper by Dumith et al. published in this issue of Preventive Medicine. METHODS: We discuss the added value of the manuscript by Dumith et al. while acknowledging some limitations of the analyses. RESULTS: We argue that there is a global and urgent need to take action against physical inactivity. CONCLUSION: The paper by Dumith et al. provides context to help build the case for better public health organization and effort to tackle physical inactivity at a worldwide level. PMID- 21645538 TI - Role of phosphoinositide 3-kinase-Akt signaling pathway in the age-related cytokine dysregulation in splenic macrophages stimulated via TLR-2 or TLR-4 receptors. AB - Age-associated defects in both B-lymphocytes and macrophages in elderly result in a reduction in the efficacy of vaccines to many Gram positive bacteria like Streptococcus pneumoniae. Splenic macrophages from aged mice have been shown to have a defect in production of pro-inflammatory cytokines (IL-6, IL-12, IL-1beta, TNF-alpha) but exhibit increased production of IL-10 upon TLR-4 ligation. Here we showed that aged macrophages demonstrate similar cytokine dysregulation phenotype upon stimulation with TLR-2 ligands, or killed S. pneumoniae. We hypothesized that an age-associated increase in activity of phosphatidyl inositol 3-kinase (PI3K)-Akt signaling pathway may be playing a causal role in the age-associated cytokine dysregulation. We found that gene expression of both the regulatory (p85beta) and the catalytic (p110delta) subunits of Class IA PI3K is higher in aged than in young splenic macrophages. The age-associated increase in the activity of PI3K was also demonstrated by an upregulation of P-Akt and its downstream target, glycogen synthase kinase-3 (GSK-3). Inhibition of PI3K enhanced induction of pro-inflammatory cytokines, by TLR-2/TLR-1, TLR-2/TLR-6 and TLR-4 ligands as well as heat killed S. pneumoniae (HKSP). Therefore, targeting PI3-Kinase could rescue cytokine dysregulation in aged macrophages and enhance the relevant pro-inflammatory cytokines needed to support B-cell activation and differentiation. PMID- 21645540 TI - The interaction between residues 62 and 193 play a key role in activity and structural stability of arginine kinase. AB - The purpose of this study is to clarify that the amino acid residues (Asp62 and Arg193) are responsible for the activity and stability of arginine kinase (AK). The amino acid residues Asp62 (D62) and Arg193 (R193) are strictly conserved in monomeric AKs and form an ion pair in the transition state analogue complex. In this research, we replaced D62 with glutamate (E) or glycine (G) and R193 with lysine (K) or glycine (G). The mutants of D62E and R193K retained almost 90% of the wild-type activity, whereas D62G and R193G had a pronounced loss in activity. A detailed comparison was made between the physic-chemical properties and conformational changes of wild-type AK and the mutants by means of ultraviolet (UV) difference and fluorescence spectra. The results indicated that the conformation of all of the mutants had been changed and the stability in a urea solution was also reduced. We speculated that the hydrogen bond and electrostatic interactions formed between residues 62 and 193 play a key role in stabilizing the structure and mediating the synergism in substrate binding of arginine kinase from greasyback shrimp (Metapenaeus ensis). PMID- 21645541 TI - Auditory frequency-following response: a neurophysiological measure for studying the "cocktail-party problem". AB - How do we recognize what one person is saying when others are speaking at the same time? The "cocktail-party problem" proposed by Cherry (1953) has puzzled scientific societies for half a century. This puzzle will not be solved without using appropriate neurophysiological investigation that should satisfy the following four essential requirements: (1) certain critical speech characteristics related to speech intelligibility are recorded; (2) neural responses to different speech sources are differentiated; (3) neural correlates of bottom-up binaural unmasking of responses to target speech are measurable; (4) neural correlates of attentional top-down unmasking of target speech are measurable. Before speech signals reach the cerebral cortex, some critical acoustic features are represented in subcortical structures by the frequency following responses (FFRs), which are sustained evoked potentials based on precisely phase-locked responses of neuron populations to low-to-middle-frequency periodical acoustical stimuli. This review summarizes previous studies on FFRs associated with each of the four requirements and suggests that FFRs are useful for studying the "cocktail-party problem". PMID- 21645542 TI - Passing the knife edge in adolescence: brain pruning and specification of individual lines of development. PMID- 21645543 TI - Effects of general and local fatigue on postural control: a review. AB - This review addresses the effects of fatiguing general muscular exercise (involving the whole body) and fatiguing local muscular exercise (involving a particular muscular group) on postural control. Short and intensive general exercise increases postural sway when the energy expenditure induced exceeds the lactate accumulation threshold. Exhaustive local exercise affects postural control when it generates a strength loss at least 25-30% of maximal voluntary contraction. Non-intensive general and local exercises can also disturb postural control when the exercise is prolonged. Both general and local exercises contribute to altering the effectiveness of sensory inputs and motor output of postural control. Different compensatory postural strategies are triggered to counteract or limit the disturbance of postural control due to the general and local muscle fatigue. Conceptual models are proposed to explain how the central contribution privileges certain sensory information and neglects and/or compensates other information and improves motor output of postural control by developing motor strategies according to the context of muscle fatigue. PMID- 21645544 TI - Recent progress in cell therapy for basal ganglia disorders with emphasis on menstrual blood transplantation in stroke. AB - Cerebrovascular diseases are the third leading cause of death and the primary cause of long-term disability in the United States. The only approved therapy for stroke is tPA, strongly limited by the short therapeutic window and hemorrhagic complications, therefore excluding most patients from its benefits. Parkinson's and Huntington's disease are the other two most studied basal ganglia diseases and, as stroke, have very limited treatment options. Inflammation is a key feature in central nervous system disorders and it plays a dual role, either improving injury in early phases or impairing neural survival at later stages. Stem cells can be opportunely used to modulate inflammation, abrogate cell death and, therefore, preserve neural function. We here discuss the role of stem cells as restorative treatments for basal ganglia disorders, including Parkinson's disease, Huntington's disease and stroke, with special emphasis to the recently investigated menstrual blood stem cells. We highlight the availability, proliferative capacity, pluripotentiality and angiogenic features of these cells and explore their present and future experimental and clinical applications. PMID- 21645545 TI - Time to blood culture positivity as a predictor of drug resistance in Acinetobacter baumannii complex bacteremia. PMID- 21645546 TI - Upregulation of heme oxygenase-1 via PI3K/Akt and Nrf-2 signaling pathways mediates the anti-inflammatory activity of Schisandrin in Porphyromonas gingivalis LPS-stimulated macrophages. AB - The lipopolysaccharide (LPS) of Porphyromonas gingivalis is thought to induce periodontitis. In this study, we isolated Schisandrin from the dried fruits of Schisandra chinensis and examined the anti-inflammatory effect of Schisandrin in macrophages stimulated with LPS from P. gingivalis. First, Schisandrin inhibited LPS-induced pro-inflammatory cytokines, including TNF-alpha, IL-1beta, and IL-6. And Schisandrin suppressed the nuclear translocation and activity of NF-kappaB and phosphorylation of IkappaBalpha in LPS-stimulated RAW 264.7 cells. Next, the presence of a selective inhibitor of HO-1 (SnPP) and a siRNA specific for HO-1 inhibited Schisandrin-mediated anti-inflammatory activity. Furthermore, Schisandrin induced HO-1 expression of RAW 264.7 cells through Nrf-2, PI3K/Akt, and ERK activation. Therefore, these results suggest that the anti-inflammatory effects of Schisandrin on P. gingivalis LPS-stimulated RAW 264.7 cells may be due to a reduction of NF-kappaB activity and induction of the expression of HO-1, leading to TNF-alpha, IL-1beta, and IL-6 down-regulation. PMID- 21645547 TI - Experimental intracerebral vaccination protects mouse from a neurotropic virus by attracting antibody secreting cells to the CNS. AB - In previous studies, we showed that intracerebrally (IC) immunized mice had antigen-specific antibodies (Abs) in cerebrospinal fluid and could survive lethal doses of transneurally spreading viruses. To better understand the mechanisms behind this, immune responses in both the central nervous system (CNS) and lymphoid organs following intracerebral immunization against pseudorabies virus (PRV) were investigated by focusing on antibody secreting cells (ASCs). IC immunized mice had significantly higher PRV-specific serum Abs and neutralizing Abs titers than SC immunized mice. Spleen and cervical lymph nodes (CLNs) of IC immunized mice produced significantly more PRV-specific Abs than that of SC immunized mice. ASCs, immunoglobulin and mRNAs of IgG, CXCL9, 10, 13 and BAFF were predominantly detected in the brain of IC immunized mice, but not in SC immunized mice. IC immunized mice (86%) survived more than subcutaneously (SC) immunized mice (33%) by suppression of virus propagation, when PRV was inoculated directly into the brain. In conclusion, IC immunization induced more effective immune responses to protect the CNS from PRV infection by attracting ASCs into the CNS and inducing much more PRV-specific serum neutralizing Abs. This approach may have important implications as a novel treatment procedure for neurotropic virus infections in both humans and animals. PMID- 21645548 TI - Application of a split luciferase complementation assay for the detection of viral protein-protein interactions. AB - Intraviral protein-protein interactions are critical for virus survival in the host. Discovery of such interactions is important to understand molecular mechanisms of viral replication and pathogenesis. The development of a cell-based assay that can be employed to examine systematically viral protein interactions is described. The method, known as the split luciferase complementation assay (SLCA), is based on the principle that N- and C-terminal domains of luciferase alone do not emit luminescence; however, if fused to interacting proteins the two non-functional halves can be brought into close enough proximity through a specific protein-protein interaction to restore the functions of the enzyme and emit detectable light. The well-studied influenza B polymerase acidic protein (PA) and basic protein 1 (PB1) interaction was used as a model system to develop the assay. Consistent with previous studies, a strong PA-PB1 interaction was demonstrated in the assay. The PA-PB1 interaction was also disrupted by single amino acid mutations in the N-terminal domain of PB1 that is responsible for binding PA. The described SLCA is highly specific and easy to perform, and thus may be useful for studying protein-protein interactions in viral diseases. PMID- 21645549 TI - An optimized sensitive method for quantitation of DNA/RNA viruses in heparinized and cryopreserved plasma. AB - Sodium heparin, an anticoagulant used widely for blood collection, has been known to inhibit DNA polymerase activity in polymerase chain reaction (PCR) assays. However, all cryopreserved plasma samples collected in the 1980s and early 1990s at the Multicenter AIDS Cohort Study were from heparin-treated blood, which poses a problem in quantifying the target nucleic acids contained in those samples by PCR assay. In this study, a nucleic acid extraction procedure was optimized to remove the heparin from extracted nucleic acids. Using this optimized method, similar human immunodeficiency virus 1 (HIV-1) and cytomegalovirus (CMV) loads of these viruses that were added to normal donor blood from ethylenediaminetetraacetic acid (EDTA), acid citrate dextrose (ACD) or sodium heparin tubes were detected by reverse transcriptase (RT) real-time PCR and real time PCR. Comparable HIV-1 and CMV loads were also detected in the blood of persons with active HIV-1 and CMV infections collected in EDTA-, ACD- or sodium heparin-treated tubes by RT real-time and real-time PCR. The findings showed that the optimized nucleic acid extraction procedure efficiently removes the heparin inhibition effect on the performance of real-time PCR. This method could be used to extract nucleic acids from archived, heparinized plasma for PCR based quantitation of target molecules. PMID- 21645550 TI - Missense mutation of the reticulon-4 receptor alters spatial memory and social interaction in mice. AB - The reticulon-4 receptor, encoded by RTN4R, limits axonal sprouting and neural plasticity by inhibiting the outgrowth of neurites. Human association studies have implicated mutations in RTN4R in the development of schizophrenia, including the identification of several rare nonconservative missense mutations of RTN4R in schizophrenia patients. To investigate the effects of missense mutation of the reticulon-4 receptor on phenotypes relevant to schizophrenia, we behaviourally characterized a novel Rtn4r mutant mouse line with an amino acid substitution (R189H) in the Nogo-66 binding site. Behavioural assays included prepulse inhibition of acoustic startle, locomotor activity, social interaction and spatial cognition. When compared with wildtype littermates, Rtn4r mutant mice exhibited greater social preference, which may reflect a social-anxyolitic effect, and a mild impairment in spatial cognition. Given the mild effect of the R189H mutation of Rtn4r on behavioural phenotypes relevant to schizophrenia, our results do not support missense mutation of RTN4R as a strong risk factor in the pathogenesis of schizophrenia. PMID- 21645551 TI - Behavioral effects of elevated expression of human equilibrative nucleoside transporter 1 in mice. AB - Adenosine concentrations are regulated by purinergic enzymes and nucleoside transporters. Transgenic mice with neuronal expression of human equilibrative nucleoside transporter 1 (hENT1) have been generated (Parkinson et al., 2009 [7]). The present study tested the hypothesis that mice homozygous and heterozygous for the transgene exhibit differences in hENT1 mRNA and protein expression, and in behavioral responses to caffeine and ethanol, two drugs with adenosine-dependent actions. Real time polymerase chain reaction (PCR) was used to identify mice heterozygous and homozygous for the transgene. Gene expression, determined by real time PCR of cDNA reverse transcribed from cerebral cortex RNA, was 3.8-fold greater in homozygous mice. Protein abundance, determined by radioligand binding assays using 0.14nM [(3)H]S-(4-nitrobenzyl)-6-thioinosine ([(3)H]NBTI), was up to 84% greater in cortex synaptosome membranes from homozygous than from heterozygous mice. In western blots with an antibody specific for hENT1, a protein of approximately 40kDa was strongly labelled in cortex samples from homozygous mice, weakly labelled in samples from heterozygous mice and absent from samples from wild type mice. In behavioral assays, transgenic mice showed a greater response to ethanol and a reduced response to caffeine than wild type littermates; however, no significant differences between heterozygous and homozygous mice were detected. These data indicate that the difference in ENT1 function between wild type and heterozygous mice was greater than that between heterozygous and homozygous mice. Therefore, either heterozygous or homozygous hENT1 transgenic mice can be used in studies of ENT1 regulation of adenosine levels and adenosine dependent behaviors. PMID- 21645552 TI - The influence of extrinsic motivation on competition-based selection. AB - The biased competition approach to visuo-spatial attention proposes that the selection of competing information is effected by the saliency of the stimulus as well as by an intention-based bias of attention towards behavioural goals. Wascher and Beste (2010) [32] showed that the detection of relevant information depends on its relative saliency compared to irrelevant conflicting stimuli. Furthermore the N1pc, N2pc and N2 of the EEG varied with the strength of the conflict. However, this system could also be modulated by rather global mechanisms like attentional effort. The present study investigates such modulations by testing the influence of extrinsic motivation on the selection of competing stimuli. Participants had to detect a luminance change in various conditions among others against an irrelevant orientation change. Half of the participants were motivated to maximize their performance by the announcement of a monetary reward for correct responses. Participants who were motivated had lower error rates than participants who were not motivated. The event-related lateralizations of the EEG showed no motivation-related effect on the N1pc, which reflects the initial saliency driven orientation of attention towards the more salient stimulus. The subsequent N2pc was enhanced in the motivation condition. Extrinsic motivation was also accompanied by enhanced fronto-central negativities. Thus, the data provide evidence that the improvement of selection performance when participants were extrinsically motivated by announcing a reward was not due to changes in the initial saliency based processing of information but was foremost mediated by improved higher-level mechanisms. PMID- 21645553 TI - Effects of the 5-HT(6) receptor agonist ST 1936 on depression- and anhedonia-like experimental models. AB - Serotonin 5-HT(6) receptor agonists and antagonists have been proposed as possible useful compounds in the treatment of psychiatric disorders such as depression. This study was aimed at characterizing ST 1936, a new 5-HT(6) receptor agonist, as a possible antidepressant/anti-anhedonic drug by studying its effects on three experimental models of depression. These models are based on the behavioral sequelae induced in rats by unavoidable stressors that result in decreased reactivity to avoidable stressors (escape deficit, ED) and an anhedonia like condition based on the disruptive effect of stress on the competence to acquire an instrumental vanilla sugar-sustained appetitive behavior (VAB). The repeated administration of ST 1936 prevented the development of ED, but did not revert a condition of chronic ED. The protective effect of ST 1936 was antagonized by co-administration of SB 271046, a 5-HT(6) receptor antagonist, indicating that the 5-HT(6) receptor stimulation is crucial for triggering a plasticity process that resulted in the prevention of ED development. ST 1936 administration in rats undergoing VAB training did not interfere with its acquisition, whereas SB 271046 administered in similar conditions prevented VAB acquisition. Moreover, ST 1936 administration in rats trained in the Y-maze while exposed to a chronic stress protocol consistently antagonized the stress disrupting effect, and also this effect was antagonized by SB 271046 coadministration. It was concluded that a tonic 5-HT(6) receptor activity was crucial for VAB acquisition, and that pharmacological stimulation of 5-HT(6) receptors reinstated a stress-reduced hedonic competence with an efficacy similar to that of classical antidepressant drugs. PMID- 21645554 TI - Early life stress decreases hippocampal BDNF content and exacerbates recognition memory deficits induced by repeated D-amphetamine exposure. AB - Adverse experiences early in life may have profound influences on brain development, for example, determining alterations in response to psychostimulant drugs, an increased risk of developing a substance abuse disorder, and individual differences in the vulnerability to neuropsychiatric disorders later in life. Here, we investigated the effects of exposure to an early adverse life event, maternal deprivation, combined with repeated d-amphetamine (AMPH) administration in adulthood, on recognition memory and brain-derived neurotrophic factor (BDNF) levels in rats' brain and serum. Rats were exposed to one of the following maternal rearing conditions from postnatal days 1 to 14: non-deprived (ND) or deprived (D). In adulthood, both groups received injections of saline (SAL) or AMPH (2.0mg/kg, i.p.) for 7 days. In Experiment I (performed 24h after the last AMPH injection), AMPH induced long-term memory (LTM) impairments in ND and D groups. The D+AMPH group also presented short-term memory (STM) impairments, indicating that the effects of AMPH on memory were more pronounced when the animals where maternally deprived. The group exposed to D+SAL (SAL) showed only LTM impairments. In Experiment II (performed 8 days after the last injection), AMPH detrimental effects on memory persisted in ND and D groups. BDNF levels were decreased in the hippocampus of D+SAL rats. In conclusion, AMPH produces severe and persistent recognition memory impairments that were more pronounced when the animals were maternally deprived, suggesting that an early adverse life event may increase the vulnerability of cognitive function to exposure to a psychostimulant later in life. PMID- 21645555 TI - Dysfunctional long-term potentiation-like plasticity in schizophrenia revealed by transcranial direct current stimulation. AB - Neural and cortical plasticity represent the ability of the brain to reorganize its function in response to a challenge. Plasticity involves changing synaptic activity and connectivity. Long-term-potentiation is one important mechanism underlying these synaptic changes. Disturbed neuronal plasticity is considered to be part of the pathophysiology of schizophrenia and has been linked to the different clinical features of this severe illness. The aim of the present study was to investigate nonfocal cortical plasticity and cortical excitability in recent-onset and multi-episode schizophrenia compared with healthy subjects. Nonfocal cortical plasticity can be induced in the motor cortex of healthy subjects with anodal transcranial direct current stimulation. Animal and human research indicates that this long-term-potentiation-like plasticity is glutamate dependent and that these plasticity shifts can last for several hours. Transcranial direct current stimulation-induced plasticity was monitored by transcranial magnetic stimulation-generated motor evoked potentials. Well characterized transcranial magnetic stimulation protocols were applied to determine the physiological basis of plasticity changes. Multi-episode schizophrenia patients showed significantly reduced long-term-potentiation-like plasticity compared to recent-onset schizophrenia patients and healthy controls. All schizophrenia patients demonstrated reduced cortical inhibition. Our results indicate that the long-term-potentiation-like plasticity deficit in schizophrenia patients is related to the disease course. Disturbances of N-methyl-d-aspartate, gamma-aminobutyric acid and dopamine receptors may account for this plasticity deficit. LTP-like plasticity deficits might be related to disturbed information processing in schizophrenia patients. PMID- 21645556 TI - Delta(9)-THC and WIN55,212-2 affect brain tissue levels of excitatory amino acids in a phenotype-, compound-, dose-, and region-specific manner. AB - Endocannabinoids are involved in excitatory neurotransmission initiated by glutamate and aspartate. The aim of the present study was to investigate the effects of the cannabinoid agonists, Delta(9)-THC and WIN55,212-2, on tissue (prefrontal cortex, dorsal striatum, nucleus accumbens, hippocampus, amygdala and hypothalamus) levels of glutamate and aspartate in two rat phenotypes, high responders (HR) and low responders (LR), differentiated according to their response to a novel environment. HR displayed increased motor activity but no difference in basal levels of glutamate and aspartate as compared to LR. Both cannabinoids increased ambulatory activity at the low doses, this effect was observed only in HR following Delta(9)-THC, but in both HR and LR following WIN55,212-2. The cannabinoids primarily increased glutamate levels in the prefrontal cortex, dorsal striatum, nucleus accumbens and hippocampus, while the high dose of WIN55,212-2 decreased glutamate levels in the amygdala and both doses in the hypothalamus; these effects appeared overall more pronounced in HR. In contrast, the cannabinoids primarily decreased aspartate levels in all brain regions, except in the dorsal striatum, where an increase was seen after both doses of Delta(9)-THC and WIN55,212-2 as well as in the nucleus accumbens after the low dose of Delta(9)-THC in HR; these effects also appeared overall more pronounced in HR. Present results show that exogenous cannabinoids affect tissue levels of glutamate and aspartate in a phenotype-, compound-, dose-, and brain region-dependent manner. PMID- 21645557 TI - Does prenatal methamphetamine exposure affect the drug-seeking behavior of adult male rats? AB - Methamphetamine (MA) is one of the most frequently used illicit drugs worldwide and also one of the most common drugs abused by pregnant women. Repeated administration of psychostimulants induces behavioral sensitization in response to treatment of the same or related drugs in rodents. The effect of prenatal MA exposure on sensitivity to drugs in adulthood is not yet fully determined. Because our most recent studies demonstrated that prenatal MA (5mg/kg) exposure makes adult rats more sensitive to acute injection of the same drug, we were interested whether the increased sensitivity corresponds with the increased drug seeking behavior. The aim of the present study was to examine the effect of prenatal MA exposure on drug-seeking behavior of adult male rats tested in the conditioned place preference (CPP). The following psychostimulant drugs were used as a challenge in adulthood: MA (5mg/kg), amphetamine (5mg/kg) and cocaine (10mg/kg). All psychostimulant drugs induced increased drug-seeking behavior in adult male rats. However, while MA and amphetamine-induced increase in drug seeking behavior did not differ based on the prenatal drug exposure, prenatally MA-exposed rats displayed tolerance effect to cocaine in adulthood. In addition, prenatally MA-exposed rats had decreased weight gain after administration of MA or amphetamine, while the weight of prenatally MA-exposed rats stayed unchanged after cocaine administration. Defecation was increased by all the drugs (MA, amphetamine and cocaine), while only amphetamine increased the tail temperature. In conclusion, our results did not confirm our hypothesis that prenatal MA exposure increases drug-seeking behavior in adulthood in the CPP test. PMID- 21645558 TI - Phasic reward responses in the monkey striatum as detected by voltammetry with diamond microelectrodes. AB - Reward-induced burst firing of dopaminergic neurons has mainly been studied in the primate midbrain. Voltammetry allows high-speed detection of dopamine release in the projection area. Although voltammetry has revealed presynaptic modulation of dopamine release in the striatum, to date, reward-induced release in awakened brains has been recorded only in rodents. To make such recordings, it is possible to use conventional carbon fibres in monkey brains but the use of these fibres is limited by their physical fragility. In this study, constant-potential amperometry was applied to novel diamond microelectrodes for high-speed detection of dopamine. In primate brains during Pavlovian cue-reward trials, a sharp response to a reward cue was detected in the caudate of Japanese monkeys. Overall, this method allows measurements of monoamine release in specific target areas of large brains, the findings from which will expand the knowledge of reward responses obtained by unit recordings. PMID- 21645559 TI - TACE cleaves neogenin to desensitize cortical neurons to the repulsive guidance molecule. AB - Neogenin is a receptor for netrins and proteins of the repulsive guidance molecule (RGM) family. It regulates several key developmental processes within the nervous system. The binding of RGMa to neogenin induces the inhibition of neurite outgrowth and the collapse of the growth cone of neurons. Here, we report that a disintegrin and metalloprotease (ADAM) transmembrane protein regulates the sensitivity of neurons to RGMa, by inducing the shedding of the ectodomain of neogenin. The extracellular domain of neogenin is directly associated with and cleaved off by the tumor necrosis factor-alpha converting enzyme (TACE), also called ADAM17. TACE is endogenously expressed in embryonic cortical neurons and regulates the cleavage of neogenin, and the inhibition of TACE in turn enhances RGMa-induced inhibition of neurite outgrowth and collapse of the growth cone. Conversely, exogenous expression of TACE abolishes the effect of RGMa. Therefore, TACE may play a role in modulating the RGM-induced repulsive behavior of neurons by regulating the expression of neogenin on the cell surface. PMID- 21645560 TI - Conditional gene expression in hepatitis C virus transgenic mice without induction of severe liver injury using a non-inflammatory Cre-expressing adenovirus. AB - We previously established inducible-hepatitis C virus (HCV) transgenic mice, which expressed the HCV gene (nucleotides 294-3435) encoding the core, E1, E2, and NS2 proteins. The expression of these proteins is regulated by the Cre/loxP system and an adenovirus vector (AdV) that expresses Cre DNA recombinase (Cre) controlled by the CAG promoter (AxCANCre). Recent studies have demonstrated that AxCANCre injection alone results in severe liver injury by induction of the adenovirus protein IX (Ad-pIX) gene. As a result, HCV protein expression in transgenic mice livers was only short-term. In contrast, the EF1alpha promoter bearing AdV induces slight Ad-pIX gene expression without inducing severe liver injury. Therefore, in the present study, we developed a Cre-expressing AdV that bears the EF1alpha promoter (AxEFCre) to express HCV protein in the transgenic mouse livers. In the non-transgenic mice injected with AxCANCre, alanine aminotransferase (ALT) levels were elevated and severe liver inflammation occurred; this was not observed in AxEFCre-injected mice. In contrast, AxEFCre injected HCV transgenic mice showed milder liver inflammatory responses that were clearly due to HCV protein expression. Moreover, the AxEFCre injection enabled the transgenic mice to persistently express HCV protein. These results indicate that use of AxEFCre efficiently promotes Cre-mediated DNA recombination in vivo without a severe hepatitis response to AdV. This inducible-HCV transgenic mouse model using AxEFCre should be useful for research on HCV pathogenesis. PMID- 21645561 TI - Sequence analysis of a Torque teno canis virus isolated in China. AB - In the present study, a total of 158 fecal samples were collected from diarrheal dogs younger than 1 year old in pet clinic in China. 20 specimens (20/158, 13%) were positive for Torque teno canis virus DNA using detection PCR. One representative positive isolate designated LDL was randomly selected, cloned and sequenced. The complete genome of the LDL Chinese strain was 2799 nucleotides in length and contains three open reading frames (ORFs), which encode 576 (ORF1), 101 (ORF2), and 243 (ORF3) aa. Compared with the human and other animal TTV genomes, the genome of the LDL strain is clearly smaller and shares 95% identity with Japanese cf-TTV10 strain (AB076002). Phylogenetic analysis showed that the present Chinese Torque teno canis virus LDL strain was also closely clustered with the previous Japanese cf-TTV10 strain, and formed a different branch together with Torque teno sus viruses 1 and 2 compared with other Torque teno viruses, Torque teno mini virus, and Torque teno midi virus. Our study demonstrated that Torque teno canis virus is present in China. PMID- 21645562 TI - Development of recombinant rabies viruses vectors with Gaussia luciferase reporter based on Chinese vaccine strain CTN181. AB - The recombinant rabies virus (RV) vectors encoding the secreted gene marker Gaussia luciferase (Gluc) were generated based on Chinese vaccine strain CTN181. Vectors included replication competent CTN-Gluc, CTN/G(Q333R)-Gluc, in which the amino acid in position 333 of glycoprotein was mutated from glutamine (Q) to arginine (R), and replication constrained CTNDeltaG-Gluc, in which the glycoprotein encoding gene (G) was deleted. The growth of recombinant RVs in transfected cells was confirmed through biochemical assays of Gluc activities. Gluc expression in recombinant CTNDeltaG-Gluc virus was highest while that in CTN/G(Q333R)-Gluc virus was lowest. The optimal time to harvest recombinant RVs was determined and the function of pathogenic and nonpathogenic rabies glycoprotein in virus recovery was examined. The addition of glycoprotein was slightly beneficial for virus recovery and the titer of rescued virus was lowered even when the amino acid in G333 position of glycoprotein was mutated from nonpathogenic Gln to pathogenic Arg. CONCLUSIONS: Viral vectors based on a human rabies vaccine strain CTN181 were successful. Gluc was useful as an in vitro gene marker for monitoring the growth of recombinant RVs iteratively in cell culture. PMID- 21645563 TI - Eighth International Nanomedicine and Drug Delivery Symposium (NanoDDS'10). PMID- 21645564 TI - Metal ion release kinetics from nanoparticle silicone composites. AB - Metal ion release kinetics from silver and copper nanoparticle silicone composites generated by laser ablation in liquids are investigated. The metal ion transport mechanism is studied by using different model equations and their fit to experimental data. Results indicate that during the first 30 days of immersion, Fickian diffusion is the dominant transport mechanism. After this time period, the oxidation and dissolution of nanoparticles from the bulk determine the ion release. This second mechanism is very slow since the dissolution of the nanoparticle is found to be anisotropic. Silver ion release profile is best described by pseudo-first order exponential equation. Copper ion release profile is best described by a second order exponential equation. For practical purposes, the in vitro release characteristics of the bioactive metal ions are evaluated as a function of nanoparticle loading density, the chemistry and the texture of the silicone. Based on the proposed two-step release model, a prediction of the release characteristics over a time course of 84 days is possible and a long-term ion release could be demonstrated. PMID- 21645565 TI - In-shell pistachio nuts reduce caloric intake compared to shelled nuts. AB - It was hypothesized that consuming in-shell pistachios, compared to shelled pistachios, causes individuals to consume less. A convenience sample of students at a mid-western university (n=140) was recruited, asking them to evaluate a variety of brands of pistachios. A survey at the end of class determined fullness and satisfaction. Subjects entering the classroom were given a 16-ounce cup and asked to self-select a portion of pistachios. Portion weight was recorded and subjects consumed pistachios at their leisure during class. At class end, pistachios remaining in the cup were weighed and total consumption by weight was determined. The caloric content of each portion was then calculated. In condition one, subjects offered in-shell pistachios consumed an average of 125 calories. In condition two, subjects offered shelled pistachios consumed an average of 211 calories; a difference of 86 calories. Subjects in condition one consumed 41% fewer calories compared to subjects in condition two (p<=.01). Fullness and satisfaction ratings were not significantly different (p>=.01). Caloric intake was influenced by the initial form of the food. The difference in calories consumed may be due to the additional time needed to shell the nuts or the extra volume perceived when consuming in-shell nuts. PMID- 21645566 TI - Cognitive advantages of chewing gum. Now you see them, now you don't. AB - The current series of experiments investigated the effects of the timing of gum chewing on cognitive function, by administering a battery of cognitive tasks to participants who chewed gum either prior to or throughout testing, and comparing their performance to that of controls who did not chew gum. Chewing gum was associated with performance advantages on multiple measures when gum was chewed for 5 min before, but not during, cognitive testing. The benefits, however, persisted only for the first 15-20 min of the testing session, and did not extend to all cognitive domains. To explain this pattern of results, it is proposed that the time-limited nature of performance benefits can be attributed to mastication induced arousal. Furthermore, the lack of improvement in cognitive function when gum is chewed throughout testing may be because of interference effects due to a sharing of resources by cognitive and masticatory processes. This dual-process mechanism is not only consistent with the outcome of present experiments but can potentially account for a wide range of findings reported in the literature. PMID- 21645567 TI - Shared and separate functions of the RAMP-based adrenomedullin receptors. AB - Adrenomedullin (AM) is a novel hypotensive peptide that exerts a variety of strongly protective effects against multiorgan damage. AM-specific receptors were first identified as heterodimers composed of calcitonin-receptor-like receptor (CLR), a G protein coupled receptor, and one of two receptor activity-modifying proteins (RAMP2 or RAMP3), which are accessory proteins containing a single transmembrane domain. RAMPs are required for the surface delivery of CLR and the determination of its phenotype. CLR/RAMP2 (AM1 receptor) is more highly AM specific than CLR/RAMP3 (AM2 receptor). Although there have been no reports showing differences in intracellular signaling via the two AM receptors, in vitro studies have shed light on their distinct trafficking and functionality. In addition, the tissue distributions of RAMP2 and RAMP3 differ, and their gene expression is differentially altered under pathophysiological conditions, which is suggestive of the separate roles played by AM1 and AM2 receptors in vivo. Both AM and the AM1 receptor, but not the AM2 receptor, are crucial for the development of the fetal cardiovascular system and are able to effectively protect against various vascular diseases. However, AM2 receptors reportedly play an important role in maintaining a normal body weight in old age and may be involved in immune function. In this review article, we focus on the shared and separate functions of the AM receptor subtypes and also discuss the potential for related drug discovery. In addition, we mention their possible function as receptors for AM2 (or intermedin), an AM-related peptide whose biological functions are similar to those of AM. PMID- 21645568 TI - RYH: a minimal peptidic sequence obtained from beta-chain hemoglobin exhibiting an antimicrobial activity. AB - Bovine hemoglobin is an animal protein described as source of bioactive peptides. Enzymatic hydrolysis of this protein results into some peptides exhibiting antimicrobial activity against Gram-positive and Gram-negative bacteria. In this study, a family of peptides from the beta chain (beta-114-145 derived peptides) obtained by peptic hydrolysis of bovine hemoglobin, was purified by reverse-phase HPLC and characterized by different analytical techniques (mass spectrometry, circular dichroism). The minimum inhibitory concentration was determined to show the antimicrobial activity of these peptides. Four bacterial strains were used: two Gram-negative (Escherichia coli and Salmonella Enteritidis) and two Gram positive strains (Listeria innocua and Micrococcus luteus). The effect of these peptides on artificial membrane was also measured. Our findings showed that the peptide beta114-145 and its peptic derivatives contain the RYH sequence. The most antimicrobial peptide is the RYH peptide which was the shortest one. PMID- 21645569 TI - A CD40 and an NCOA5 gene polymorphism confer susceptibility to psoriasis in a Southern European population: a case-control study. AB - Recent genome-wide association studies of many complex diseases have successfully identified novel susceptibility loci, with many of them shared by multiple disease-associated pathways. The genes CD40 and nuclear receptor coactivator 5 (NCOA5), located in a 400-kb region surrounding CD40, have been reported to be associated with increased risk for rheumatoid arthritis and other autoimmune diseases. We hypothesized that those genes may also have a role in psoriasis (PS), an autoimmune, chronic inflammatory skin disease. In a case-control study, 198 patients with PS and 400 controls were genotyped for 2 single nucleotide polymorphisms (SNPs) of the CD40 and NCOA5 genes located on chromosome 20q.12 q13.12. Here, we demonstrate for the first time the association of both SNPs with susceptibility to PS, thus suggesting a putative key role of both genes in multiple autoimmune diseases. Alleles G and C of the CD40 rs4810485 and NCOA5 rs2903908 SNPs, respectively, were more common in individuals with PS than in controls (p = 0.03, odds ratio [OR] = 1.42, 95% confidence interval [95% CI] 1.05 1.95 and p = 0.000 003, OR = 1.93, 95% CI 1.47-2.55, respectively). The identification of shared genetic susceptibility loci may provide insight into our understanding of the pathophysiology of autoimmune diseases. PMID- 21645570 TI - Transmission of HLA-DP variants from parents to children with B-cell precursor acute lymphoblastic leukemia: log-linear analysis using the case-parent design. AB - Childhood B-cell precursor acute lymphoblastic leukemia (BCP ALL) is usually initiated in utero and is thought to progress to overt leukemia under the influence of delayed exposure to a common infection. Based on the hypothesis that polymorphic HLA-DP variants can restrict T-cell responses to infection, we previously compared DP supertype frequencies in BCP ALL patients with that of unrelated newborn controls. We reported that the DP2 supertype was associated with susceptibility, whereas DP1 was associated with protection. However, the association of genetic variants in children with early-onset diseases such as ALL may be a proxy for parental effects. Here we examine whether maternal DP1 and DP2 are associated with BCP ALL by fitting log-linear models in a combined series of family triads (both parents and case child) and dyads (1 parent and case child; n = 571) in comparison with similar models in non-BCP leukemia (n = 198). We report no evidence of maternal DP1 or DP2 associations with BCP ALL, but we did identify suggestive evidence of maternal undertransmission of the infrequent supertypes DP11 and DP15. Although these results require confirmation, they suggest that DP11 and DP15 may be protective or that there is transmission ratio distortion of these supertypes in BCP ALL. PMID- 21645571 TI - SLAM and DC-SIGN measles receptor polymorphisms and their impact on antibody and cytokine responses to measles vaccine. AB - BACKGROUND: Despite the use of measles vaccine, measles virus continues to circulate and cause severe disease. Immune responses to the measles vaccine are variable between individuals, with up to 10% failing to produce a sufficient protective response post-vaccination. Signaling lymphocyte activation molecule (SLAM) and dendritic cell-specific intercellular adhesion molecule-3 grabbing nonintegrin (DC-SIGN; CD209) are specific measles receptors: SLAM binds and permits entry of the virus into the cell, DC-SIGN acts as an attachment receptor, increasing viral binding efficiency and transmission. Genetic variations in these receptor genes may alter measles vaccine antibody and cellular responses. METHODS: In 12-month-old infants from Perth, Western Australia after their first measles vaccine dose as part of the combination measles-mumps-rubella (MMR) vaccine, 7 SLAM and DC-SIGN polymorphisms were genotyped and associations were investigated with measles IgG antibody levels and in vitro measles cytokine responses. RESULTS: The DC-SIGN promoter variant -336C/T was associated with overall IFN-gamma responses after measles stimulation (P=0.002) and three DC-SIGN polymorphisms (-336C/T, -139C/T and -871C/T) were associated with the proportion of cytokine non-responders to measles (P=0.001, P=0.021 and P=0.036, respectively). However, no associations were found between the DC-SIGN or SLAM polymorphisms and measles IgG antibody levels. CONCLUSIONS: The results suggest that DC-SIGN -139C/T, -336C/T and -871C/T polymorphisms may modulate cytokine (but not antibody) responses to the measles component of MMR vaccine. Furthermore, contrasting previous studies, SLAM polymorphisms do not appear to affect measles antibody or cytokine responses in this cohort. PMID- 21645572 TI - Escheriosome mediated cytosolic delivery of Candida albicans cytosolic proteins induces enhanced cytotoxic T lymphocyte response and protective immunity. AB - Protection against intracellular fungal infections, in a manner similar to viral challenges necessitates activation of both humoral and cell mediated immune responses in unison. Most of the presently available antigen delivery vehicles including egg phosphatidyl-choline (egg-PC) liposomes, can evoke mainly humoral immune responses in the immunized animals. Keeping this fact into consideration, we earlier developed Escherichia coli membrane lipid vesicles (escheriosomes) and demonstrated that escheriosomes successfully fuse with the plasma membrane of macrophages ensuing in effective cytoplasmic delivery of entrapped antigen, a pre requisite for inducing CD8(+) T cell response against antigens. In the present study, we report the ability of escheriosomes encapsulated Candida albicans (C. albicans) cytosolic antigens (cAg), to generate protective immunity against systemic C. albicans infection in BALB/c mice. The immunization schedule using escheriosome encapsulated cAg induced strong antigen-specific CD8(+) T-cell responses, which were markedly higher than that observed in mice immunized with IFA-antigen emulsion, or antigen encapsulated in egg PC liposomes. Interestingly, immunization with cAg delivered in escheriosomes was also successful in complete elimination of C. albicans infection in Balb/c mice. The study suggests that escheriosomes may function as a novel immunoadjuvants and emerge as an effective tool for generating protective immunity against C. albicans infection. PMID- 21645573 TI - Pneumococcal conjugate vaccination at birth in a high-risk setting: no evidence for neonatal T-cell tolerance. AB - Concerns about the risk of inducing immune deviation-associated "neonatal tolerance" as described in mice have restricted the widespread adoption of neonatal vaccination. The aim of this study was to demonstrate the immunological feasibility of neonatal pneumococcal conjugate vaccination (PCV) which could potentially protect high-risk infants in resource poor countries against severe pneumococcal disease and mortality in the early critical period of life. Papua New Guinean infants were randomized to be vaccinated with the 7-valent PCV (7vPCV) at birth, 1 and 2 months (neonatal group, n=104) or at 1, 2 and 3 months of age (infant group, n=105), or to not receive 7vPCV at all (control group, n=109). Analysis of vaccine responses at 3 and 9 months of age demonstrated persistently higher type-1 (IFN-gamma) and type-2 (IL-5 and IL-13) T-cell responses to the protein carrier CRM(197) and IgG antibody titres to 7vPCV serotypes in children vaccinated with 7vPCV according to either schedule as compared to unvaccinated children. In a comprehensive immuno-phenotypic analysis at 9 months of age, no differences in the quantity or quality of vaccine-specific T cell memory responses were found between neonatal vaccinations versus children given their first PCV dose at one month. Hospitalization rates in the first month of life did not differ between children vaccinated with PCV at birth or not. These findings demonstrate that neonatal 7vPCV vaccination is safe and not associated with immunological tolerance. Neonatal immunisation schedules should therefore be considered in high-risk areas where this may result in improved vaccine coverage and the earliest possible protection against pneumococcal disease and death. PMID- 21645574 TI - Immunisation with the glycolytic enzyme enolase confers effective protection against Candida albicans infection in mice. AB - Candida albicans is an opportunistic human fungal pathogen that continues to be a leading cause of candidal infections in immunocompromised hosts. Enolase, an important glycolytic enzyme located on the cell wall of C. albicans, was cloned, purified, and characterized by molecular cloning, affinity chromatography and Western blotting. C57BL/6J mice were immunized with recombinant enolase subcutaneously every two weeks, and the protective effect against systemic challenge evaluated by fungal burdens in target organs, titres of specific antibodies to enolase, and by levels of Th1/2 cytokines in serum. After challenge with C. albicans strains SC5314 and 3630, fungal burdens in the liver, kidney, brain, spleen and lung were significantly decreased in immunized mice. Histopathological assessment demonstrated that enolase protected the tissue structure, and decreased the infiltration of inflammatory cells. The titres of enolase-specific IgG1 and IgG2a in the immune serum reached up to 1:51200. Furthermore, opsonization with immune serum resulted in enhanced killing of both 3630 and SC5314 by murine neutrophils. Levels of IL-12 and IL-8 in the immune serum increased, whereas the concentration of the Th2 cytokine, IL-10, was significantly higher in immunized mice compared to the control group. It was concluded that recombinant enolase effectively protected mice against disseminated candidiasis, and may be a promising target for vaccination against different strains of C. albicans. PMID- 21645575 TI - Neutralizing antibody but not hemagglutination antibody provides accurate evaluation for protective immune response to H5N1 avian influenza virus in vaccinated rabbits. AB - In order to develop an animal model and an assay method to evaluate protective immune response to H5N1 avian influenza vaccination, H5N1 avian influenza vaccine was prepared. New Zealand rabbits were assigned to receive two doses of vaccine with different hemagglutinin (HA) dosage. The sera from vaccinated rabbits was evaluated to determine antibody titer and specificity using different tested methods including hemagglutination inhibition assay (HI), neutralizing assay (NT), cross-HI assay, cross-single immunodiffusion assay and cross-neutralization assay. The titer of HI antibody from rabbits immunized with different doses of HA were no less than 1:40 among groups 14 days after the first immunization. Whereas the NT antibody titer was less than 1:10 among groups 14 days after the first immunization. NT antibodies can be detected 14 days after the second immunization in rabbits immunized at HA doses higher than 6 MUg, and the NT antibody titers were equal to or higher than 1:40. A good concentration-dependent NT antibody response can be detected in the vaccinated rabbits 14 days after the second immunization, and in contrast, no concentration-dependent relationship can be seen for HA antibody. The cross-HI test showed sera from vaccinated rabbits could cross react with influenza A H5N1 virus with the titers higher than 1:40. No cross reaction among different types (influenza A/H1N1 virus, influenza A/H3N2 virus, influenza B virus and influenza A/H5N1 virus) can be detected in the sera using the single immunodiffusion assay and using NT antibody test. This showed NT antibody test was demonstrated as a more accurate assay method for evaluating vaccination and quality of the vaccine than HI antibody test. PMID- 21645576 TI - Evaluating the sensitization potential of surfactants: integrating data from the local lymph node assay, guinea pig maximization test, and in vitro methods in a weight-of-evidence approach. AB - An integral part of hazard and safety assessments is the estimation of a chemical's potential to cause skin sensitization. Currently, only animal tests (OECD 406 and 429) are accepted in a regulatory context. Nonanimal test methods are being developed and formally validated. In order to gain more insight into the responses induced by eight exemplary surfactants, a battery of in vivo and in vitro tests were conducted using the same batch of chemicals. In general, the surfactants were negative in the GPMT, KeratinoSens and hCLAT assays and none formed covalent adducts with test peptides. In contrast, all but one was positive in the LLNA. Most were rated as being irritants by the EpiSkin assay with the additional endpoint, IL1-alpha. The weight of evidence based on this comprehensive testing indicates that, with one exception, they are non sensitizing skin irritants, confirming that the LLNA tends to overestimate the sensitization potential of surfactants. As results obtained from LLNAs are considered as the gold standard for the development of new nonanimal alternative test methods, results such as these highlight the necessity to carefully evaluate the applicability domains of test methods in order to develop reliable nonanimal alternative testing strategies for sensitization testing. PMID- 21645577 TI - Treatment of motor symptoms in advanced Parkinson's disease: a practical approach. AB - Patients with advanced Parkinson's disease (PD) are known to develop motor complications after a few years of levodopa (L-dopa) therapy. Motor fluctuations develop with increasing severity of the disease, owing to loss of dopaminergic neurons and loss of the buffering capacity of the neurons to fluctuating dopamine levels. Dyskinesias develop as a result of pulsatile stimulation of the receptors and alterations in neuronal firing patterns. L-dopa remains the gold standard medication for the treatment of patients with advanced PD. However, once motor complications on L-dopa therapy emerge, clinicians may add on other classes of antiparkinsonian drugs such as dopamine agonists, catechol-O-methyl transferase inhibitors (COMTIs) or monoamine oxidase type B inhibitors (MAOBIs). The individualisation of the treatment seems to be the key for the best approach of advanced PD patients. The present review provides the most important current clinical data in the pharmacological treatment of motor symptoms in advanced PD and provides the clinician a simple algorithm in order to determine the best suitable treatment to advanced parkinsonian patients. PMID- 21645578 TI - Toxicological studies on 2,4,6-tribromoanisole. AB - TBA, or 2,4,6-tribromoanisole, is a musty-smelling metabolite of 2,4,6 tribromophenol that is used as a flame retardant and an antifungal agent for wooden pallets and packaging materials. The compound can impart its peculiar, often offensive, odor on product packaging to the concern of consumers for the safety of the package contents. These studies were conducted to evaluate the safety of TBA to humans ingesting products tainted with TBA. In addition to the 28-day oral study, a bacterial reverse mutation study was conducted, and confirmed that TBA was not mutagenic. To evaluate oral safety, TBA was evaluated in single dose and 5-day and 4-week repeated dose oral toxicity studies in rats. The test article, administered in single gavage doses of 2000, 5000 and 7500 mg/kg body weight (bw), in 5 daily repeated doses of 1000, 2000 or 3000 mg/kg bw/day or in 28 daily oral gavage doses of 0 001, 0.01, 100, and 1000 mg/kg bw/day did not result in any deaths. Also, the single and repeat dose studies resulted in no significant differences between control and treated groups on body weight gain, food consumption, clinical observations, blood biochemical values, and hematology findings. Treatment-related adverse findings were only detected in male rats during repeated dose studies and were associated with high plasma concentrations of TBA. The test article-related finding of hyaline droplets in the cortical tubular epithelium of kidneys was associated with increases in alpha(2 MU)-globulin content in the kidneys as indicated by the intensity of immunohistochemical staining. These findings were correlated with an increased weight of kidneys in males administered 1000mg/kgbw/day for 28days. Chemical induction of hyaline droplets containing alpha(2MU)-globulin in the renal proximal tubule is a process unique to the male rat and is not relevant for human risk assessment. Findings of increased liver weight with minimal centrilobular to diffuse hepatocellular hypertrophy in males treated with TBA at 1000mg/kg bw/day for 28days were considered to be an adaptive metabolic response to xenobiotic administration. The increased volume of urine, noted in both males and females treated with 1000mg/kg bw/day was considered adaptive and necessary to excrete the high xenobiotic burden resulting from TBA administration. TBA appeared to be highly bioavailable since high concentrations of TBA were detected in plasma, at 1, 4 and 8h after administration of TBA at 100 and 1000mg/kg bw for 1 and 28days. Levels were dose-related but did not clarify the course of TBA elimination with time after administration. These studies indicate that TBA, administered orally to rats, produced male rat-specific, treatment-related toxicity at the highest orally administered dose in repeated dose (5-day at 3000mg/kg bw and 28-day at 1000mg/kg bw) studies. Therefore, the NOAEL for the 28-day oral study was determined to be 1000mg/kg bw/day for the rat. PMID- 21645579 TI - Fucoidan, a sulfated polysaccharide from brown algae, against myocardial ischemia reperfusion injury in rats via regulating the inflammation response. AB - The aim of the study was to determine the effects of fucoidan on rat myocardial ischemia-reperfusion (I/R) model and elucidate the potential mechanisms. Myocardial I/R injury was induced by the occlusion of left anterior descending coronary artery for 30 min followed by reperfusion for 2h. After 2h reperfusion, hemodynamics parameters were detected. Blood samples were collected to determine serum levels of tumor necrosis factor-alpha (TNF-alpha) and interleukin 6, 10 (IL 6, 10). Hearts were harvested to assess histopathological changes, infarct size (IS), and the content of myeloperoxidase (MPO). The expression of high-mobility group box 1 (HMGB1), phosphor-IkappaB-alpha and phosphor-nuclear factor kappa B (NF-kappaB) were assayed by western blot. Compared with control group, treatment with fucoidan improved left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP) and the contractility index (P<0.05, P<0.01). Fucoidan reduced the myocardial IS, the levels of TNF-alpha and IL-6, and the activity of MPO (P<0.05, P<0.01). Fucoidan down-regulated the expression of HMGB1, phosphor-IkappaB-alpha and NF-kappaB, but increased the content of IL 10 when compared with control (P<0.05, P<0.01). Besides, the infiltration of polymorph nuclear leukocytes (PMNs) and histopathological damages in myocardium were decreased in fucoidan treated groups (PMNs, P<0.05, P<0.01). These findings revealed that the administration of fucoidan could regulate the inflammation response via HMGB1 and NF-kappaB inactivation in I/R-induced myocardial damage. PMID- 21645580 TI - Oral subchronic and genotoxicity studies conducted with the amino acid, L glutamine. AB - L-Glutamine is an abundantly occurring amino acid that serves numerous nutritional and physiological functions. It has current and potential applications as a therapeutic agent, dietary supplement, food ingredient, and in animal nutrition. To assess the safety of supplemental L-glutamine, a bacterial reverse mutation assay, in vitro chromosomal aberration assay, and a 13-week toxicity study were conducted. L-Glutamine showed no mutagenic activity in the bacterial reverse mutation assay, and did not induce chromosomal aberrations in Chinese hamster lung fibroblast cells in the in vitro chromosomal aberration assay. In the 13-week toxicity study, Sprague-Dawley rats (10/sex/group) were fed diets containing 0, 0.5, 2.5, or 5.0% L-glutamine. No deaths occurred, and no significant differences in body weights, body weight gains, ophthalmological findings, urinalysis parameters, or organ weights were observed between L glutamine-fed rats and their respective controls. No toxicologically relevant effects on hematological or blood biochemical parameters were observed. Macroscopic and microscopic effects occurred at low frequency but were not associated with a dose-response relationship. Based on the results of the study, the no-observed-adverse-effect-level was determined to be 5.0% L-glutamine in the diet, the highest concentration tested (equivalent to 3832 and 4515 mg/kg body weight/day in male and female rats, respectively). PMID- 21645581 TI - Designing and surface modification of zinc oxide nanoparticles for biomedical applications. AB - The present study aimed to work out a simple and high yield procedure for the immobilization of beta galactosidase on bioaffinity support, concanavalin A (Con A) layered zinc oxide nanoparticles (ZnO-NP). Thermogravimetric analysis of bioaffinity support revealed 4% loss in weight at 600 degrees C whereas its thermal decomposition was observed at 530 degrees C by differential thermal analysis. No significant change was noticed in the band intensity of pUC19 plasmid after its treatment with Con A layered ZnO-NP. Comet assay further exhibited negligible change in tail length of comet after treating the lymphocytes by bioaffinity matrix. The bioaffinity matrix binds 89% of the enzyme activity. Atomic force microscopy analysis showed that the prepared matrix has an advantageous microenvironment and large surface area for binding significant amount of the enzyme. The functional groups present in native and parent compound were monitored by Fourier transform-infrared spectroscopy. Michaelis constant, K(m) was 2.38 and 5.88 mM for free and immobilized beta galactosidase, respectively. V(max) for the soluble and immobilized enzyme was 0.520 mM/min and 0.460 mM/min, respectively. Concanavalin A layered ZnO-NP bound beta galactosidase exhibited a shift in the temperature-optima and retained nearly 86% activity even after its 6th repeated use. PMID- 21645582 TI - Influences of methamphetamine-induced acute intoxication on urinary and plasma metabolic profiles in the rat. AB - Methamphetamine (MA) is an illicit psychostimulant, and its abuse has become an international public health problem. MA intoxication can cause life-threatening hyperthermia, renal and liver failure, cardiac arrhythmias, and neurological damage. To investigate the relationship between the underlying mechanism of such intoxication and metabolic networks, mass spectrometry-based metabolomics experiments were performed on Sprague-Dawley rats treated with MA at 10mgkg(-1)h( 1) for 4h. Using a combination of gas chromatography-time-of-flight mass spectrometry and capillary electrophoresis-tandem mass spectrometry, global and targeted analyses were performed on biological samples collected during 0-24 and 72-96h (for urine), and at 24 and 96h (for plasma) after the last drug administration. Body temperature and plasma biochemical parameters were also measured to detect abnormal reactions in neuronal and other several tissues. 5 Oxoproline, saccharic acid, uracil, 3-hydroxybutyrate (3-HB), adipic acid, glucose, glucose 6-phosphate, fructose 1,6-bisphosphate, and tricarboxylic acid (TCA) cycle intermediates, such as fumarate, were proposed as potential biomarkers related to MA-induced intoxications. In particular, the observation of decreased TCA cycle intermediates and 3-HB and increased glucose suggested that high doses of MA inhibit biogenic energy production by glycolysis, oxidative phosphorylation via the TCA cycle, and the beta-oxidation of fatty acids. These results may provide not only a clue to clarify the underlying mechanism of diverse intoxication effects, but also biological fluid-based diagnostic and forensic methods with which to objectively demonstrate intoxication without directly determining the drug. PMID- 21645583 TI - Protective effects of ion-imprinted chitooligosaccharides as uranium-specific chelating agents against the cytotoxicity of depleted uranium in human kidney cells. AB - Occupational internal contamination with depleted uranium (DU) compounds can induce radiological and chemical toxicity, and an effective and specific uranium chelating agent for clinical use is urgently needed. The purpose of this study was to investigate whether a series of synthesized water-soluble metal-ion imprinted chitooligosaccharides can be used as uranium-specific chelating agents, because the chitooligosaccharides have excellent heavy metal ion chelation property and the ion-imprinting technology can improve the selective recognition of template ions. DU-poisoned human renal proximal tubule epithelium cells (human kidney 2 cells, HK-2) were used to assess the detoxification of these chitooligosaccharides. The DU-chelating capacity and selectivity of the chitooligosaccharides were determined by inductively coupled plasma-mass spectrometry (ICP-MS). Cell viability, cellular accumulation of DU, membrane damage, DNA damage, and morphological changes in the cellular ultrastructure were examined to assess the detoxification of these chitooligosaccharides. The results showed that the Cu2+-imprinted chitooligosaccharides, especially the Cu2+ imprinted glutaraldehyde-crosslinked carboxymethyl chitooligosaccharide (Cu-Glu CMC), chelated DU effectively and specifically, and significantly reduced the loss of cell viability induced by DU and reduced cellular accumulation of DU in a dose-dependent manner, owing to their chelation of DU outside cells and their prevention of DU internalization. The ultrastructure observation clearly showed that Cu-Glu-CMC-chelated-DU precipitates, mostly outside cells, were grouped in significantly larger clusters, and they barely entered the cells by endocytosis or in any other way. Treatment with Cu-Glu-CMC also increased the activity of antioxidant enzymes, and reduced membrane damage and DNA damage induced by DU oxidant injury. Cu-Glu-CMC was more effective than the positive control drug, diethylenetriaminepentaacetic acid (DTPA), in protection of HK-2 cells against DU cytotoxicity, as a result of its chelation of UO22+ to prevent the DU internalization and its antioxidant activity. PMID- 21645584 TI - Dehydroepiandrosterone in nails of infants: a potential biomarker of intrauterine responses to maternal stress. AB - Easily accessible biomarkers for fetal stress biology are lacking. We here explore whether quantification of major fetal steroids, dehydroepiandrosterone (DHEA) or DHEA sulfate (DHEAS), with liquid chromatography/tandem mass spectrometry in infant nails is a tool to assess fetal stress biology in response to maternal stressful life events during pregnancy. Sufficient nail (>= 1 mg) was available from 80 infants (93% of those providing samples). The concentration of DHEA, but not DHEAS, was increased in infants of mothers with stressful life events during pregnancy (DHEA: F1,41=6.105, P=0.018; DHEAS: F1,77=0.767, P=0.384). DHEA concentrations were not related to maternal stress before pregnancy (F1,41=0.010, P=0.922). Infant nail DHEA may be a fetal biological correlate of intrauterine exposure to maternal stress. The method promises the first non-invasive retrospective biomarker for intrauterine stress biology, opening new ways for research and clinical applications in fetal medicine, endocrinology, obstetrics, gynecology, and for understanding the developmental origins of health and disease. PMID- 21645585 TI - Resting posterior minus frontal EEG slow oscillations is associated with extraversion and DRD2 genotype. AB - The agency facet of extraversion has been hypothesized to be based on individual differences in dopamine activity. Recent work suggests that resting posterior minus frontal electroencephalographic (EEG) slow oscillations (delta, theta) is both consistently associated with extraversion and sensitive to dopamine D2 receptor antagonist-induced changes in dopaminergic activity. Here we examine for the first time the interrelations between polymorphisms of the dopamine D2 receptor (DRD2) gene (rs1800497 [previously termed TAQ1A], rs1076560, rs1799732 [ 141C Ins/Del]), extraversion and resting posterior minus frontal (Pz-Fz) slow oscillations. As predicted, we found an association between DRD2 and resting Pz Fz slow oscillations in a sample of 141 individuals participating in an eyes closed resting EEG session. Moreover, we replicated the association between extraversion and Pz-Fz slow oscillations. Our findings strongly suggest that the posterior-frontal distribution of slow oscillations constitutes a useful brain based intermediate phenotype for investigating the dopaminergic basis of extraversion. PMID- 21645586 TI - Prediction of metabolic pathways from genome-scale metabolic networks. AB - The analysis of a variety of data sets (transcriptome arrays, phylogenetic profiles, etc.) yields groups of functionally related genes. In order to determine their biological function, associated gene groups are often projected onto known pathways or tested for enrichment of known functions. However, these approaches are not flexible enough to deal with variations or novel pathways. During the last decade, we developed and refined an approach that predicts metabolic pathways from a global metabolic network encompassing all known reactions and their substrates/products, by extracting a subgraph connecting at best a set of seed nodes (compounds, reactions, enzymes or enzyme-coding genes). In this review, we summarize this work, while discussing the problems and pitfalls but also the advantages and applications of network-based metabolic pathway prediction. PMID- 21645587 TI - GnRH receptor mutations in isolated gonadotropic deficiency. AB - GnRH and its receptor GnRHR are key regulators of the hypothalamo-pituitary axis. They modulate the secretion of LH and FSH gonadotropins and therefore, the development and maturation of gonads in fetal life as well as after birth. Congenital functional defect of this axis results in isolated hypogonadotropic hypogonadism (IHH). Several natural mutations causing IHH without anosmia have now been identified in GnRHR or GnRH genes. These mutations inactivate GnRHR or its ligand function and cause highly variable phenotypes, ranging from partial to complete gonadotropic deficiencies. The present review describes the published natural GnRHR mutations and tries to correlate them with the corresponding phenotypes according to the different steps of the GnRH system development. PMID- 21645588 TI - Male Internet addicts show impaired executive control ability: evidence from a color-word Stroop task. AB - This study investigated the executive control ability of male students with Internet addiction disorder (IAD) by recording event-related brain potentials (ERP) during a color-word Stroop task. Seventeen IAD and 17 male normal university students participated. Behavior results showed that IAD students were associated with longer reaction time and more response errors in incongruent conditions than the control group. ERP results revealed that participants with IAD showed reduced medial frontal negativity (MFN) deflection in incongruent conditions than the control group. Both of the behavioral performance and ERP results indicate that people with IAD show impaired executive control ability than the normal group. PMID- 21645589 TI - Neurological Soft Signs and Corpus Callosum morphology in schizophrenia. AB - OBJECTIVE: Neurological Soft Signs (NSS) have been found to be more prevalent in schizophrenic patients. A breakdown in intracortical functional connectivity, including interhemispheric communication, has been suggested in the pathogenesis of schizophrenia. Indeed, problems with interhemispheric information transfer via the Corpus Callosum (CC) have been documented in schizophrenics. Our study goal was to relate NSS to CC morphology. METHODS: CC Magnetic Resonance Imaging (MRI) measurements were collected from 29 right-handed male schizophrenia inpatients. NSS were evaluated employing the Neurological Evaluation Scale (NES). We examined the scores obtained from the NES total and the three NES subscales: Integrative Sensory Function, Motor Coordination, and Sequencing Of Complex Motor Acts. We compared CC morphology of patients with "high" NSS with that of patients with "low" NSS. Correlation analyses were performed to further clarify the relationship between CC size, NSS, and total lifetime antipsychotic consumption. RESULTS: Patients with "high" scores at the Sequencing Of Complex Motor Acts subscale showed a smaller CC rostral body, whereas patients with "high" scores at the Integrative Sensory Function subscale showed a smaller CC splenium. For both the NES total and the Sequencing Of Complex Motor Acts subscale, "high" scores were accompanied by an increase of the CC genu. Correlation analyses revealed a significant inverse correlation between the CC rostral body size and the Sequencing Of Complex Motor Acts subscale score. In addition, a significant positive correlation was shown between the CC genu size and both the NES total and the Sequencing Of Complex Motor Acts subscale scores. The presence of NSS and the accompanying CC structural abnormalities were independent on antipsychotic treatment. CONCLUSIONS: Our data provide evidence for an association between NSS and CC morphology and further support the hypothesis of a disturbed interhemispheric functional connectivity in schizophrenia. PMID- 21645590 TI - Cell survival or cell death: differential vulnerability of long descending and thoracic propriospinal neurons to low thoracic axotomy in the adult rat. AB - Previous studies show that most short thoracic propriospinal (TPS; T5-T7) and long descending propriospinal tract (LDPT; C4-C6) neurons are lost following low thoracic spinal cord contusion injury (cSCI), as assessed by retrograde labeling with fluorogold (FG). Gene microarray and terminal deoxynucleotidyl transferase dUTP nick end (TUNEL)/caspase-3 immunolabeling indicate that post-axotomy cell death may be responsible for the observed decrease in number of labeled TPS neurons post cSCI. Yet, no indications of post-axotomy cell death are evident within LDPT neurons following the same injury. The present experiments were devised to understand this difference. We assessed the number and size of LDPT and TPS neurons at different time points, retrogradely labeling these neurons with FG prior to delivering a moderate low-thoracic cSCI or after they were axotomized by a complete low-thoracic spinal transection. Counts of FG-filled TPS and LDPT cells indicate a large loss of both neuronal populations 2 weeks post cSCI. Propriospinal neurons in other animals were retrogradely labeled with dextran tetramethyl rhodamine prior to cSCI and tissue was processed for detection of TUNEL- or caspase-3-positive profiles at chronic times post injury. Our overall findings confirm that cell death post injury is the major factor responsible for the loss of TPS neurons during the acute period post cSCI, and that little post-axotomy cell death occurs in LDPT neurons during the first 2 months after the same injury. After chronic axotomy retrograde transport is impaired in LDPT neurons, but can be reinitiated by re-axotomy. Our results also indicate that FG is metabolized/lost from retrogradely labeled neurons at increasing survival times, and that this process appears to be accelerated by injury. PMID- 21645591 TI - Positive emotional learning is regulated in the medial prefrontal cortex by GluN2B-containing NMDA receptors. AB - In rats, hedonic ultrasonic vocalizations (USVs) is a validated model of positive affect and is best elicited by rough-and-tumble play. Here we report that modulation of GluN2B-containing NMDA receptors (NMDAR) in the medial prefrontal cortex (MPFC) is involved in positive emotional learning. Rough and tumble play increased both GluN1 and GluN2B NMDAR subunit mRNA and protein levels in the frontal cortex. GLYX-13, a GluN2B-preferring, NMDAR glycine-site partial agonist (1 mg/kg, i.v.) significantly increased positive emotional learning whereas the GluN2B receptor-specific antagonist, ifenprodil (10 mg/kg, i.p.), inhibited positive emotional learning. Animals selectively bred for low rates of hedonic USVs were returned to wild-type levels of positive emotional learning following GLYX-13 treatment. MPFC microinjections of GLYX-13 (0.1-10 MUg/side) significantly increased rates of positive emotional learning. Thus GluN2B containing NMDARs may be involved in positive emotional learning in the MPFC by similar mechanisms as spatial/temporal learning in the hippocampus. PMID- 21645592 TI - Song sharing and singing strategies in the winter wren Troglodytes troglodytes. AB - The winter wren is a common forest bird living in groups of few adjacent neighbours during the breeding season. Inside each group, males vocally interact in the context of both territorial holding and sexual competition, forming a complex communication network. To study this network, we first analysed song type and syllable repertoires within and between distinct groups. We found a limited number of song types highly stereotyped in length, syntax and syllable composition, frequently shared among neighbours. Between groups, song type and syllable repertoires sharing decreased with increasing distance at a higher rate for song types than for syllables. Then, with continuous recordings, we focused on the dynamics of acoustic interactions between neighbours. We showed that male winter wrens can differentially use their song type repertoire (non-matching strategy), overlap their neighbours and modulate their singing rhythm producing longer inter-song intervals with no change in song length during acoustic interactions. PMID- 21645593 TI - Transdermal delivery of selegiline from alginate-Pluronic composite thermogels. AB - The present work was carried out to design a practical, controlled-release transdermal system for selegiline using thermosensitive hydrogels. The copolymers of alginate and Pluronic F127 (PF127) were used to design thermogels by either physical blending (A+P) or chemical grafting (AP). The thermogels were characterized in terms of the sol-gel temperature, scanning electron microscopy (SEM), degradation ratio, and skin permeation behavior. The chemical grafting of alginate to PF127 could delay the sol-gel temperature from 24.1 to 30.4 degrees C, which is near the temperature of the skin surface. The gelling temperature of the physical mixture of alginate and PF127 (A+P) did not significantly differ. The porosity of the A+P structure was greater compared to that of the AP structure. AP thermogels were regularly degraded, with 60% of the gel matrix remaining after a 48-h incubation. PF127 and A+P hydrogels showed almost no degradation. The results of skin permeation across porcine skin and nude mouse skin suggested that the thermogels could produce sustained selegiline release, with AP showing the most-sustained permeation. AP hydrogels exhibited linear permeation properties for the transdermal delivery of selegiline. Inter-subject variations in skin permeation were reduced by incorporation of the thermogel. Such a thermosensitive hydrogel can be advantageous as a topical therapeutic formulation for selegiline. PMID- 21645594 TI - Enhanced anti-tumor effect of 9-nitro-camptothecin complexed by hydroxypropyl beta-cyclodextrin and safety evaluation. AB - The aim of this study was to evaluate the safety and anti-tumor effect of 9-nitro camptothecin/hydroxypropyl-beta-cyclodextrin (9-NC/HP-beta-CD) complex on tumor bearing mice. The in vitro anti-tumor activity was tested by MTT assay. Our study revealed that the 9-NC/HP-beta-CD complex showed significant anti-tumor activity towards Skov-3, MCF-7, HeLa and S180 cell lines with IC(50) values of 0.24 +/- 0.09, 0.59 +/- 0.20, 0.83 +/- 0.11, and 6.30 +/- 2.42 MUg/ml, respectively, significantly superior to the free 9-NC. The in vivo therapeutic efficacy was investigated in ICR mice bearing mouse sarcoma S180. Both the high (3mg/kg) and low (1mg/kg) doses of 9-NC/HP-beta-CD complex demonstrated high inhibition ratio of tumor growth (>75%). The subacute toxicity test was performed by measuring the body weight, histopathology, blood cell counts and clinical chemistry parameters (total bilirubin, alanine transferase, aspartate transferase, blood urea nitrogen and creatinine), and the results indicated the good safety profile of the complex. Taken together, the results suggested that the 9-NC complexed in HP-beta CD, instead of dissolved in the organic solvent, presented significant anti-tumor activity and low toxicity for the treatment of cancer. PMID- 21645595 TI - Characterisation of the interactive properties of microcrystalline cellulose carboxymethyl cellulose hydrogels. AB - Combinations of microcrystalline cellulose (MCC) and sodium carboxymethyl cellulose (Na-CMC) are commonly used as stabilising agents and suspending agents in pharmaceutical formulations. This paper is based on a study of the interactions that take place during the process of hydrogel formation, break down, and recovery. Also considered is the binding that occurs between the MCC and the Na-CMC. Avicel RC 591, a processed mixture of MCC and Na-CMC, is one of the more commonly used commercial suspending agents for aqueous compositions. Avicel RC 591 is used as an effective, blended stabilising agent. In this study, the contributions made by each of the components of Avicel RC 591 have been rationalised by monitoring the behaviour of the individual components in Avicel RC 591 suspensions or solutions. The hydrogels that are formed by Avicel RC 591 and by their laboratory formulated equivalent, which is spray dried (MCC+Na-CMC), have been characterised by confocal microscopy scanning electron microscopy and by dynamic light scattering. A 3D network structure that is formed by the MCC, in Avicel RC 591 is visualised. This network is supported by hydrogen bonding and by ionic interactions among and between the MCC, the Na-CMC and water. The strength of the network determines the physical properties of the hydrogel system, as seen in the rheological behaviour. PMID- 21645596 TI - Enhanced dissolution and bioavailability of biochanin A via the preparation of solid dispersion: in vitro and in vivo evaluation. AB - The present study aimed to improve the bioavailability of biochanin A, a poorly soluble bioflavonoid, via the preparation of solid dispersion (SD) using Solutol HS15 and HPMC 2910. Solubility of biochanin A was enhanced by 8-60 folds as the drug-carrier ratio was increased in SDs. Furthermore, compared to pure biochanin A or physical mixture (PM), SDs significantly improved the dissolution rate and the extent of drug release. Particularly, SDs (Drug:Solutol HS15:HPMC 2910=1:5:5 or 1:10:10) achieved the rapid and complete drug release (approximately 100% within 1h) at pH 6.8. The XRD patterns indicated that SDs might enhance the solubility of biochanin A by changing the drug crystallinity to amorphous state in addition to the solubilizing effect of hydrophilic carriers. The improved dissolution of biochanin A via SD formulation appeared to be well correlated with the enhanced oral exposure of biochanin A in rats. After an oral administration of SD (Drug:Solutol HS15:HPMC 2910=1:10:10), C(max) and AUC of biochanin A were increased by approximately 13 and 5 folds, respectively, implying that SDs could be effective to improve the bioavailability of biochanin A. In conclusion, solid dispersion with Solutol HS15 and HPMC 2910 appeared to be promising to improve the dissolution and oral exposure of biochanin A. PMID- 21645597 TI - Chitosan-modified poly(D,L-lactide-co-glycolide) nanospheres for plasmid DNA delivery and HBV gene-silencing. AB - Gene silencing using small interfering RNA (siRNA) has several potential therapeutic applications. In the present study, we investigated nanoparticles (NS) formulated using the biodegradable polymer, poly(D,L-lactide-co-glycolide) (PLGA) for plasmid DNA (pDNA) delivery. A cationic polymer, Chitosan (CHS), was incorporated in the PLGA matrix to improve pDNA loading efficiency and cellular uptake ability. PLGA-CHS NS were prepared by a spontaneous emulsion diffusion (SED) method, and various formulation factors were investigated. Spherical nanoparticles with particle size of around 60 nm were obtained under optimum formulation condition. The effectiveness of pDNA-loaded PLGA-CHS nanoparticles in expressing the indicative enhanced Green Fluorescent Protein (eGFP) and in slicing Hepatitis B virus (HBV) gene were examined in HepG2.2.15 cells. CHS modified PLGA NS exhibited much higher loading efficiency than unmodified PLGA NS. CHS-PLGA NS showed a positive zeta potential, while plain-PLGA NS were negatively charged. EGFP expression studies by observation with confocal leaser scanning microscopy (CLSM) indicated that pDNA-loaded CHS-PLGA NS were more effectively taken up by the cells than plain-PLGA NS. The corresponding results showed that the HBV gene-silencing efficiency of CHS-PLGA NS was higher than those of plain-PLGA NS and naked pDNA. Thus, CHS-PLGA NS containing pDNA could provide an effective pDNA delivery system in vitro, showing that such an approach could be useful in the treatment of viral diseases in vivo. PMID- 21645598 TI - Encapsulation enhancement and stabilization of insulin in cationic liposomes. AB - The purpose of this study was to enhance encapsulation efficiency and sustained release delivery for parenteral administration of a protein drug. To reduce the administration frequency of protein drugs, it is necessary to develop sustained delivery systems. In this study, protein drug-loaded cationic liposomes were formulated with dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), dioleoyl-3 trimethylammonium-propane (DOTAP), and cholesterol (CH) at a molar ratio of DOPE/DOTAP/CH of 2/1.5/2. Five mol% of distearoylphosphatidyl ethanolamine polyethylene glycol (DSPE-PEG) was added prior to encapsulation of the drug into liposomes. Insulin was chosen as a model protein drug and encapsulation efficiency was evaluated in various liposomes with and without DSPE-PEG. Scanning electron microscopy was used to examine the insulin-loaded cationic liposomes. Structural analysis was performed using spectropolarimetry. Additionally, the stability and cytotoxicity of insulin-loaded cationic liposomes were evaluated. Liposomes coated with DSPE-PEG showed higher insulin encapsulation efficiency than did those without DSPE-PEG, but not significantly. Moreover, among the liposomes coated with DSPE-PEG, those hydrated with 10% sucrose showed higher encapsulation efficiency than did liposomes hydrated in either phosphate-buffered saline or 5% dextrose. In vitro release of insulin was prolonged by cationic liposomes. Our findings suggest that cationic liposomes may be a potential sustained-release delivery system for parenteral administration of protein and peptide drugs to prolong efficacy and improve bioavailability. PMID- 21645599 TI - New clopidogrel napadisilate salt and its solid dispersion with improved stability and bioequivalence to the commercial clopidogrel bisulphate salt in beagle dogs. AB - The purpose of this study was to develop a novel clopidogrel napadisilate-loaded solid dispersion with improved stability and bioequivalence to the clopidogrel bisulphate-loaded commercial product. Clopidogrel napadisilate prepared in this study appeared as a white crystalline powder unlike clopidogrel base. However, this salt did not improve the solubility of clopidogrel, even with improved stability compared to clopidogrel bisulphate. To improve the solubility of clopidogrel napadisilate, a novel clopidogrel napadisilate-loaded solid dispersion was prepared by the spray-drying technique using HPMC and colloidal silica, and the physicochemical properties, dissolution and bioavailability in beagle dogs were evaluated compared to the clopidogrel bisulphate-loaded commercial product. The solid dispersion composed of clopidogrel napadisilate, HPMC and colloidal silica at a weight ratio of 11.069/3/3.5 improved solubility by 6.5-fold compared to clopidogrel napadisilate, even if it did not improve drug solubility compared to clopidogrel bisulphate. However, unlike clopidogrel bisulphate, this formulation improved the stability of clopidogrel. Furthermore, the clopidogrel napadisilate solid dispersion-loaded tablet showed similar dissolution to the clopidogrel bisulphate-loaded commercial product and was bioequivalent to the commercial product in beagle dogs. Thus, this clopidogrel napadisilate-loaded solid dispersion could be a promising candidate for improving the stability and bioavailability of clopidogrel. PMID- 21645600 TI - Near-infrared spectroscopy (NIRS) and chemometric analysis of Malaysian and UK paracetamol tablets: a spectral database study. AB - The influx of medicines from different sources into healthcare systems of developing countries presents a challenge to monitor their origin and quality. The absence of a repository of reference samples or spectra prevents the analysis of tablets by direct comparison. A set of paracetamol tablets purchased in Malaysian pharmacies were compared to a similar set of sample purchased in the UK using near-infrared spectroscopy (NIRS). Additional samples of products containing ibuprofen or paracetamol in combination with other actives were added to the study as negative controls. NIR spectra of the samples were acquired and compared by using multivariate modeling and classification algorithms (PCA/SIMCA) and stored in a spectral database. All analysed paracetamol samples contained the purported active ingredient with only 1 out of 20 batches excluded from the 95% confidence interval, while the negative controls were clearly classified as outliers of the set. Although the substandard products were not detected in the purchased sample set, our results indicated variability in the quality of the Malaysian tablets. A database of spectra was created and search methods were evaluated for correct identification of tablets. The approach presented here can be further developed as a method for identifying substandard pharmaceutical products. PMID- 21645601 TI - Quantification of polymorphic impurity in an enantiotropic polymorph system using differential scanning calorimetry, X-ray powder diffraction and Raman spectroscopy. AB - The ability to detect and quantify polymorphism of pharmaceuticals is critically important in ensuring that the formulated product delivers the desired therapeutic properties because different polymorphic forms of a drug exhibit different solubilities, stabilities and bioavailabilities. The purpose of this study is to develop an effective method for quantitative analysis of a small amount of one polymorph within a binary polymorphic mixture. Sulfamerazine (SMZ), an antibacterial drug, was chosen as the model compound. The effectiveness and accuracy of powder X-ray diffraction (PXRD), Raman microscopy and differential scanning calorimetry (DSC) for the quantification of SMZ polymorphs were studied and compared. Low heating rate in DSC allowed complete transformation from Form I to Form II to take place, resulting in a highly linear calibration curve. Our results showed that DSC and PXRD are capable in providing accurate measurement of polymorphic content in the SMZ binary mixtures while Raman is the least accurate technique for the system studied. DSC provides a rapid and accurate method for offline quantification of SMZ polymorphs, and PXRD provides a non-destructive, non-contact analysis. PMID- 21645602 TI - Protection by low-dose kanamycin against noise-induced hearing loss in mice: dependence on dosing regimen and genetic background. AB - We recently demonstrated that sub-chronic low-dose kanamycin (KM, 300 mg/kg sc, 2*/day, 10 days) dramatically reduces permanent noise-induced hearing loss (NIHL) and hair cell loss in 1 month old CBA/J mice (Fernandez et al., 2010, J. Assoc. Res. Otolaryngol. 11, 235-244). Protection by KM remained for at least 48 h after the last dose, and appeared to involve a cumulative effect of multiple doses as part of a preconditioning process. The first month of life lies within the early 'sensitive period' for both cochlear noise and ototoxic injury in mice, and CBA/J mice appear exquisitely vulnerable to noise during this period (Ohlemiller et al., 2011; Hearing Res. 272, 13-20). From our initial data, we could not rule out 1) that less rigorous treatment protocols than the intensive one we applied may be equally-or more-protective; 2) that protection by KM is tightly linked to processes unique to the sensitive period for noise or ototoxins; or 3) that protection by KM is exclusive to CBA/J mice. The present experiments address these questions by varying the number and timing of fixed doses (300 mg/kg sc) of KM, as well as the age at treatment in CBA/J mice. We also tested for protection in young C57BL/6J (B6) mice. We find that nearly complete protection against at least 2 h of intense (110 dB SPL) broadband noise can be observed in CBA/J mice at least for ages up to 1 year. Reducing dosing frequency to as little as once every other day (a four-fold decrease in dosing frequency) appeared as protective as twice per day. However, reducing the number of doses to just 1 or 2, followed by noise 24 or 48 h later greatly reduced protection. Notably, hearing thresholds and hair cells in young B6 mice appeared completely unprotected by the same regimen that dramatically protects CBA/J mice. We conclude that protective effects of KM against NIHL in CBA/J mice can be engaged by a wide range of dosing regimens, and are not exclusive to the sensitive period for noise or ototoxins. While we cannot presently judge the generality of protection across genetic backgrounds, it appears not to be universal, since B6 showed no benefit. Classical genetic approaches based on CBA/J * B6 crosses may reveal loci critical to protective cascades engaged by kanamycin and perhaps other preconditioners. Their human analogs may partly determine who is at elevated risk of acquired hearing loss. PMID- 21645603 TI - Ethnopharmacology, food production, nutrition and biodiversity conservation: towards a sustainable future for indigenous peoples. AB - ETHNOPHARMACOLOGICAL RELEVANCE: It is becoming increasingly clear that ethnopharmacology cannot be disassociated from food production, human nutrition and the conservation of the biodiversity that constitutes its resource base. AIM OF THE STUDY: This paper aims to provide a perspective of ethnopharmacology that explicitly extends the range of disciplines it covers so as to embrace food and nutrition and the biodiversity basis, both wild and agricultural, and also places it in the context of the dramatic changes to our planet that we are experiencing during a period of rapid global change and the impacts that these changes are having on human health and nutrition and on its resource base. MATERIALS AND METHODS: A review is made of recent initiatives and developments that show linkages between ethnopharmacology, agriculture, food production, nutrition and biodiversity conservation. RESULTS: Ethnopharmacology, biodiversity, agriculture, food and nutrition are inextricably linked but suffer from compartmentalization and a lack of communication which have to be overcome if progress is to be made. Fortunately, a convergence of interest between the agricultural biodiversity and the biodiversity conservation sectors has emerged in recent years and there is an increased appreciation of the need to adopt a wider approach to human nutrition than the conventional agricultural model allows; there is also a greater awareness of the important role played by diversity of crops, especially local species, and consumption of wild species in achieving balanced nutrition. An increased recognition of the key role of local communities in managing agricultural biodiversity is evident. While ethnopharmacologists have expressed concern at the relentless loss of biodiversity, there has been little direct involvement but it is perhaps now time to consider a more proactive role. Attention is also drawn to the need to assess the implications of global change for ethnopharmacology. CONCLUSIONS: Ethnopharmacologists need to take much more cognizance of the fate of the resource base - the plants, animals and microorganisms - and of the actions being undertaken under the auspices of treaties, such as the Conservation on Biological Diversity and its Global Strategy for Plant Conservation, and the International Treaty on Plant Genetic Resources for Food and Agriculture, to counter its degradation and loss. In addition, they should play a more active role in determining the actions needed to assess, monitor and conserve the resource base. Although it has been suggested that the 'golden days' of ethnopharmacology may be over, it is proposed that by embracing the challenges of broadening the remit so as to include the health aspects of wild biodiversity employed in nutrition, a new 'golden age' beckons. The paper concludes with some suggestions for action. PMID- 21645604 TI - Acute and sub-acute toxicity of the methanolic extract of Pteleopsis hylodendron stem bark. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Pteleopsis hylodendron is one of the most popular medicinal plants in Cameroon where it is used to treat measles, chickenpox, sexually transmitted diseases, female sterility, liver and kidney disorders as well as dropsy. To date there is no documented evidence corroborating its safety. This study thus aimed to determine the toxicity profile of the methanolic extract of Pteleopsis hylodendron. MATERIALS AND METHODS: The acute and sub-chronic toxicity of the methanolic extract of Pteleopsis hylodendron were investigated by employing established methods. The acute toxicity study was done by administering single doses (2-8 g/kg body weight) of plant extract to adult mice. For the sub chronic toxicity study, doses (85-680 mg/kg bw) of plant extract were administered daily to adult rats during 28 days after which the effect on organs, the hematological and biochemical parameters was assessed. RESULTS: In mice, single oral administrations of the methanolic extract of Pteleopsis hylodendron caused dose-dependent general behaviour adverse effects and mortality. The LD50 values were 3.00 and 3.60 g/kg bw for males and females respectively. In rats, daily single oral doses of the methanolic extract of Pteleopsis hylodendron provoked significant (p < 0.05) growth retardation in rats at all tested doses after 28 days of dosing. Haematological parameters showed a significant decrease in white blood cells count and significant increases red blood cells count; irrespective of the sex, all biochemical parameters studied, except triglycerides significantly (p < 0.001) increased with dose. However, a dose-dependent significant (p < 0.007) increase in HDL was observed only in male rats. Increases in liver enzymes (ALT and AST), proteins and creatinine levels correlate the observed histopathological damages (i.e. inflammation and vascular congestions) in the liver and kidneys. CONCLUSIONS: The overall results of this study indicate that the methanolic extract of Pteleopsis hylodendron stem bark possesses hepatotoxic and nephrotoxic effects at doses >= 85 mg/kg bw, suggesting that this plant should be used with caution. PMID- 21645605 TI - Uterine contraction induced by Tanzanian plants used to induce abortion. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Women in Tanzania use plants to induce abortion. It is not known whether the plants have an effect. AIMS OF STUDY: Collect data on plant use in relation to induced abortion and test the effect of plant extracts on uterine contraction. MATERIALS AND METHODS: During interviews with traditional birth attendants and nurses, plants were identified. Cumulative doses of plant extracts were added to rat uterine tissue in an organ bath, and the force and frequency of contractions recorded. Acetylcholine was used as positive control. RESULTS: 21 plant species were tested for effect on uterine contraction. 11 species increased the force of contraction, and 12 species the frequency of contractions. The strongest contractions comparable to the maximum response obtained with acetylcholine were obtained with extracts of Bidens pilosa, Commelina africana, Desmodium barbatum, Manihot esculenta, Ocimum suave, Oldenlandia corymbosa and Sphaerogyne latifolia. 7 species increased both the force and frequency of contractions. CONCLUSION: Several of the plant species induced strong and frequent contractions of the uterus, and can be used to induce an abortion. PMID- 21645606 TI - Neuroprotective effects of Eleutherococcus senticosus bark on transient global cerebral ischemia in rats. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Eleutherococcus senticosus Maxim., classified into the family of Araliaceae, is used in a variety of diseases in traditional Korean medicine including ischemic heart disease. AIM OF THE STUDY: To determine the neuroprotective effects of Eleutherococcus senticosus on global cerebral ischemia. MATERIALS AND METHODS: A four-vessel occlusion (4-VO) rat model was used to evaluate the potential protective effects against transient global cerebral ischemia ethanol extracts of Eleutherococcus senticosus was orally administered at doses of 3, 30, and 300 mg/kg twice at times of 0 and 90 min after reperfusion. The effects on memory deficit were investigated by using a Y maze neurobehavioral test after brain ischemia, and the effects on hippocampal neuronal damage were measured 7 days after ischemia. The expressions of glial fibrillary acid protein (GFAP), CD11b antibody (OX-42), and cyclooxygenase-2 (COX 2) were investigated by immunohistochemistry. RESULTS: Oral administration of Eleutherococcus seticosus at 30, 100 and 300 mg/kg significantly reduced hippocampal CA1 neuronal death by 3.5%, 25.9% and 53.1%, respectively, compared with a vehicle-treated group. Oral administration of Eleutherococcus senticosus at 300 mg/kg inhibited 81.9% of the decrease in spontaneous alternation induced by 4-VOin the Y-maze test, and also attenuated ischemia-induced activation of COX 2, GFAP and OX-42 in the hippocampal CA1 region. CONCLUSION: Eleutherococcus senticosus protects delayed neuronal death in the CA1 region of the hippocampus against global cerebral ischemia in rats with the recovery of spatial memory, which can be considered as the normal functioning of the hippocampus. Regarding the immunohistochemical study, the effect of Eleutherococcus senticosus may be attributable to its anti-inflammatory properties through the inhibition of COX-2 expression, microglia and astrocyte expression. PMID- 21645607 TI - The prevalence of herbal medicine home use and concomitant use with pharmaceutical medicines in Jamaica. AB - ETHNOPHARMACOLOGICAL RELEVANCE: The work described in this paper aimed to study the prevalence of herbal medicine use in treating illness and concomitant use with pharmaceutical medicines in Jamaica. MATERIALS AND METHODS: A survey using a structured questionnaire was administered by a trained interviewer to randomly selected adults in systematically selected households within randomly selected urban and rural clusters. Categorical data analysis was performed using Stata version 10 software. RESULTS: 91.4%(372/407) of selected people agreed to participate. 72.6%(270/372) self-medicated with herbs within the previous year. Commonly treated were illnesses of the respiratory system (RS, 77.8%(210/270)), gastro-intestinal tract (GIT, 53.3%(144/270)) and health maintenance using tonics (29.6%(80/270)). 26.7%(72/270) of respondents used pharmaceuticals concomitantly with medicinal plants. Commonly treated were illnesses of the RS (20.4%(55/270)), GIT (13.7%(37/270)) and hypertension (10.0%(27/270)). 19.4% (14/72) of physicians knew of such practices. There was significant association of herb use with/without drugs with age (p<0.001), employment status (p<0.001), religion (p=0.004), gender (p=0.02) and educational level (p=0.031). Thus prevalence of herb use alone was greatest amongst people aged 35-44 and 45-54 years; those employed; Rastafarians; those without health insurance; males and people who had completed secondary education. Whilst prevalence of concomitant herb-drug use was greater amongst people aged 65 years and older; those retired; those of religions other than Rastafarians and Christians, females and people who had attained primary education and below. CONCLUSIONS: Self-medication with herbs in Jamaica is highly prevalent and highest for self-limiting conditions of the RS, GIT and health maintenance with tonics. Concomitant herb and drug use is highest for self limiting conditions of the RS, GIT and hypertension, and the use of combined therapy highlights the need for investigations on potential drug-herb interactions. Physicians have limited awareness and knowledge of such concomitant usage, further highlighting the need for increased dialogue with patients, knowledge of medicinal plants and their uses and a heightened pharmacovigilance to avoid adversities that may arise from potential drug-herb interactions. PMID- 21645608 TI - The efficacy of Prosopis glandulosa as antidiabetic treatment in rat models of diabetes and insulin resistance. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Diabetes mellitus is rampantly increasing and the need for therapeutics is crucial. In recognition of this, untested antidiabetic agents are flooding the market. DiaviteTM which is a product consisting solely of the dried and ground pods of Prosopis glandulosa (Torr.) [Fabaceae] is currently marketed as a food supplement with glucose stabilizing properties. However, these are anecdotal claims lacking scientific evidence. The aim of this study was to determine the efficacy of Prosopis glandulosa as an antidiabetic agent. MATERIALS AND METHODS: Male Wistar rats were rendered (a) type 1 diabetic after an intraperitoneal injection of STZ (40 mg/kg) and (b) insulin resistant after a 16 week high caloric diet (DIO). Zucker fa/fa ZDF rats were used in a pilot study. Half of each group of animals was placed on Prosopis glandulosa treatment (100mg/kg/day) for 8 weeks and the remaining animals served as age-matched controls. At the time of sacrifice, blood was collected for glucose and insulin level determination, the pancreata of the STZ rats were harvested for histological analysis and cardiomyocytes prepared from the DIO and Zucker fa/fa hearts for determination of insulin sensitivity. RESULTS: Type 1 diabetic model: Prosopis glandulosa treatment resulted in significant increased insulin levels (p<0.001), which was accompanied by a significant decrease in blood glucose levels (p<0.05). Additionally, Prosopis glandulosa treatment resulted in increased small beta-cells (p<0.001) in the pancreata. The body weight of the STZ animals decreased significantly after STZ injection, with Prosopis glandulosa treatment partially preventing this. Zucker fa/fa rats: Prosopis glandulosa treatment significantly reduced fasting glucose levels (p<0.01) and improved IPGTT, when comparing treated to untreated animals. DIO insulin resistant model: Prosopis glandulosa treatment resulted in an increased basal (p<0.01) and insulin stimulated (p<0.05) glucose uptake by cardiomyocytes prepared from this group. CONCLUSIONS: The present study showed that Prosopis glandulosa treatment moderately lowers glucose levels in different animal models of diabetes, stimulates insulin secretion, leads to the formation of small beta-cells and improves insulin sensitivity of isolated cardiomyocytes. PMID- 21645609 TI - In vitro screening of synergistic ascorbate-drug combinations for the treatment of malignant mesothelioma. AB - Malignant mesothelioma (MMe) is a lethal tumor arising from the mesothelium of serous cavities as a result of exposure to asbestos. Current clinical standards consist of combined treatments, but an effective therapy has not been established yet and there is an urgent need for new curative approaches. Ascorbate is a nutrient that is also known as a remedy in the treatment of cancer. In the present study, we have tested the cytotoxicity of ascorbate to MMe cells in combination with drugs used in MMe therapy, such as cisplatin, etoposide, gemcitabine, imatinib, paclitaxel, and raltitrexed, as well as with promising antitumor compounds like taurolidine, alpha-tocopherol succinate, and epigallocatechin-3-gallate (EGCG). Dose-response curves obtained for each compound by applying the neutral red uptake (NRU) assay to MMe cells growing in vitro, allowed to obtain IC50 values for each compound used singularly. Thereafter, NRU data obtained from each ascorbate/drug combination were analyzed through Tallarida's isobolograms at the IC50 level (Tallarida, 2000), revealing synergistic interactions for ascorbate/gemcitabine and ascorbate/EGCG. These results were further confirmed through comparisons between theoretical additivity IC50 and observed IC50 from fixed-ratio dose-response curves, and over a broad range of IC levels, by using Chou and Talalay's combination index (Chou and Talalay, 1984). Synergistic interactions were also shown by examining apoptosis and necrosis rates, using the caspase 3 and lactic dehydrogenase assays, respectively. Hence, data indicate that ascorbate/gemcitabine and ascorbate/EGCG affect synergistically the viability of MMe cells and suggest their possible use in the clinical treatment of this problematic cancer. PMID- 21645610 TI - Evaluation of the multipotent character of human adipose tissue-derived stem cells isolated by Ficoll gradient centrifugation and red blood cell lysis treatment. AB - In the present study, the multipotent potential of two differential isolated human adipose-derived stem cell (hADSC) populations was evaluated. More specifically, hADSC isolated by means of classical Ficoll (F) gradient centrifugation were compared to hADSC isolated by means of red blood cell (RBC) lysis treatment and subsequent cultivation as 3D spheres. No significant difference in the genotypic expression of the multipotent markers Oct-4, Sox-2, Nanog, Klf-4 and cMyc could be observed between both isolation methods. Upon adipogenic and osteogenic differentiation, both hADSC populations showed lipid droplet accumulation and mineral deposition, respectively. Although, a more pronounced mineral deposition was observed in hADSC-RBC, suggesting a higher osteogenic potential. Upon exposure to keratinogenic media, both hADSC populations expressed the keratinocyte markers filaggrin and involucrin, evidencing a successful keratinogenic differentiation. Yet, no differences in expression were observed between the distinctive isolation procedures. Finally, upon exposure to neurogenic differentiation media, a significant difference in marker expression was observed. Indeed, hADSC-RBC only expressed vimentin and nestin, whereas hADSC-F expressed vimentin, nestin, NF-200, MBP and TH, suggesting a higher neurogenic potential. In summary, our data suggest that the choice of the most efficient isolation procedure of hADSC depends on the differentiated cell type ultimately required. PMID- 21645611 TI - Nimesulide interaction with membrane model systems: are membrane physical effects involved in nimesulide mitochondrial toxicity? AB - Nimesulide (NIM), a widely used nonsteroidal anti-inflammatory drug (NSAID), is known to interfere with mitochondrial physiology and to cause idiosyncratic hepatotoxicity. In this study, we characterized the effects of NIM on the physical properties of membrane models containing the main phospholipid classes of the inner mitochondrial membrane: phosphatidylcholine (PC), phosphatidylethanolamine (PE) and cardiolipin (CL). NIM binding/incorporation was observed with the mitochondrial membrane mimicking model composed of dioleoyl PC (DOPC), dioleoyl PE (DOPE) and tetraoleoyl CL (TOCL) at a 1:1:1M ratio, as well as an increase of membrane permeability, monitored by calcein release, and an increase of lipid disorder, evaluated by fluorescence anisotropy of DPH-PA. Consistently, DSC thermograms of dipalmitoyl PC (DPPC) and a mixture of dipalmitoyl PE (DPPE) and TOCL (7:3 M ratio) showed a NIM-induced decrease of the cooperativity of the phase transition and a shift of the DPPC endotherm to lower temperatures. On the other hand, (31)P NMR studies with the ternary lipid model indicated a stabilizing effect of NIM on the lipid bilayer structure. Quenching of the fluorescent probes DPH and DPH-PA revealed a peripheral insertion of NIM in the hydrophobic portion of the bilayer. Our data indicate that NIM may influence mitochondria physiological processes by interfering with membrane structure and dynamics. The relevance of these findings will be discussed in terms of the reported NIM effects on mitochondria transmembrane potential, protonophoresis, and induction of the permeability transition pore. PMID- 21645612 TI - Time-dependent modulation of cyp1a gene transcription and EROD activity by musk xylene in PLHC-1 and RTG-2 fish cell lines. AB - Musk xylene (MX) is a common synthetic nitromusk fragrance. Its high release in aquatic environments and evidence of bioaccumulation in biota suggest that it could have serious toxicological consequences for aquatic ecosystems. However, not much data is available on cellular pathways and mechanisms of toxicity in aquatic organisms. The aim of the present study was to investigate the interaction of MX with CYP1A by looking at gene transcription and EROD activity in two fish cell lines: PLHC-1 and RTG-2. Time-dependent (6 and 24 h) exposure experiments with three doses of MX (2, 4 and 20 MUM) were performed also with co exposure to B(a)P. Low cytotoxicity was observed in both cell lines. Reduction of cyp1a gene transcription was observed after 6 h with full dose-dependent recovery in 24 h in RTG-2 and partial recovery in PLHC-1. EROD activity was inhibited after 6 and 24 h of exposure except in PLHC-1 at 6 h at the two higher doses. MX did not alter CYP1A induction by B(a)P at gene transcription. A dose and time dependent induction of GST activity was observed in PLHC-1 cells exposed to MX. These findings suggest that distinct signalling pathways not mediated by AhR and distinct regulatory mechanisms by CYP1A inducers are likely. PMID- 21645613 TI - Repeated recall of learned immunosuppression: evidence from rats and men. AB - Akin to other physiological responses, the immune system can be modified, via Pavlovian or behavioral conditioning. It is unknown, however, whether and to what extent learned immune responses can be repeatedly recalled over time. Here we demonstrate in both rats and humans that repeated contingent pairing of a novel taste (conditioned stimulus, CS) together with the immunosuppressive drug cyclosporine A as unconditioned stimulus (US) leads to the acquisition of a learned immunosuppression. Sole presentation of the CS caused a significant inhibition of interleukin (IL)-2 and interferon (IFN)-gamma production by rat splenic T cells and human peripheral T lymphocytes, closely mimicking the effect of the drug. More importantly, a comparable suppression of cytokine production was also observed after a second, unreinforced exposure to the CS that was separated from the first evocation by an interval of 6 (rats) or 11 (humans)days, respectively. Together, our findings demonstrate that a learned immunosuppression can be repeatedly recalled in both animals and humans, which is an important prerequisite for the implementation of conditioning paradigms as supportive therapy. PMID- 21645614 TI - Industrial and technical workers are not the only workers exposed to solvents. PMID- 21645616 TI - Dynamic mechanical thermal analysis of hypromellose 2910 free films. AB - It is common practice to coat oral solid dosage forms with polymeric materials for controlled release purposes or for practical and aesthetic reasons. Good knowledge of thermo-mechanical film properties or their variation as a function of polymer grade, type and amount of additives or preparation method is of prime importance in developing solid dosage forms. This work focused on the dynamic mechanical thermal characteristics of free films of hypromellose 2910 (also known as HPMC), prepared using three grades of this polymer from two different manufacturers, in order to assess whether polymer chain length or origin affects the mechanical or thermo-mechanical properties of the final films. Hypromellose free films were obtained by casting their aqueous solutions prepared at a specific concentrations in order to obtain the same viscosity for each. The films were stored at room temperature until dried and then examined using a dynamic mechanical analyser. The results of the frequency scans showed no significant differences in the mechanical moduli E' and E" of the different samples when analysed at room temperature; however, the grade of the polymer affected material transitions during the heating process. Glass transition temperature, apparent activation energy and fragility parameters depended on polymer chain length, while the material brand showed little impact on film performance. PMID- 21645617 TI - Human down syndrome cell adhesion molecules (DSCAMs) are functionally conserved with Drosophila Dscam[TM1] isoforms in controlling neurodevelopment. AB - Drosophila Down syndrome cell adhesion molecule (Dscam) potentially produces more than 150,000 cell adhesion molecules that share two alternative transmembrane/juxtamembrane (TM) domains, which dictate the dendrite versus axon subcellular distribution and function of different Dscam isoforms. Vertebrate genomes contain two closely related genes, DSCAM and DSCAM-Like1 (DSCAML1), which do not have extensive alternative splicing. We investigated the functional conservation between invertebrate Dscams and vertebrate DSCAMs by cross-species rescue assays and found that human DSCAM and DSCAML1 partially, but substantially, rescued the larval lethality of Drosophila Dscam mutants. Interestingly, both human DSCAM and DSCAML1 were targeted to the dendrites in Drosophila neurons, had synergistic rescue effects with Drosophila Dscam[TM2], and preferentially rescued the dendrite defects of Drosophila Dscam mutant neurons. Therefore, human DSCAM and DSCAML1 are functionally conserved with Drosophila Dscam[TM1] isoforms. PMID- 21645618 TI - Identification and characterization of two arylalkylamine N-acetyltransferases in the yellow fever mosquito, Aedes aegypti. AB - In this study we provide a molecular and biochemical identification of two arylalkylamine N-acetyltransferases (aaNAT) from Aedes aegypti mosquitoes. N acetyldopamine, the enzyme product of aaNAT, was detected in Ae. aegypti, indicating the presence of an aaNAT in this mosquito. A BLAST search of the Ae. aegypti genome, using sequence information from an activity-verified Drosophila aaNAT, identified thirteen putative aaNAT sequences sharing 13-48% sequence identity with the Drosophila enzyme. Eight of the thirteen putative aaNAT proteins were expressed using a bacterial expression system. Screening of purified recombinant proteins against 5-hydroxytryptamine, dopamine, methoxytryptamine, norepinephrine, octopamine, tryptamine, and tyramine substrates, established that two of the putative aaNATs are active to the tested arylalkylamines. We therefore named them aaNAT1 and 2, respectively. Analysis of the transcriptional profiles of the two aaNAT genes from Ae. aegypti revealed that aaNAT1 is more abundant in the whole body of larvae and pupae, and aaNAT2 is more abundant in the head of adult mosquitoes. Based on their substrate and transcriptional profiles, together with previous reports from other insects, we suggest that the two aaNATs play diverse roles in Ae. aegypti, with aaNAT1 primarily involved in sclerotization and aaNAT2 mainly in neurotransmitter inactivation. Our data provide a beginning to a more comprehensive understanding of the biochemistry and physiology of aaNATs from the Ae. aegypti and serve as a reference for studying the aaNAT family of proteins from other insect species. PMID- 21645619 TI - IL-1beta is induced in reactive astrocytes in the somatosensory cortex of rats with genetic absence epilepsy at the onset of spike-and-wave discharges, and contributes to their occurrence. AB - Interleukin (IL)-1beta plays a crucial role in the mechanisms of limbic seizures in rodent models of temporal lobe epilepsy. We addressed whether activation of the IL-1beta signaling occurs in rats with genetic absence epilepsy (GAERS) during the development of spike-and-wave discharges (SWDs). Moreover, we studied whether inhibition of IL-1beta biosynthesis in GAERS could affect SWD activity. IL-1beta expression and glia activation were studied by immunocytochemistry in the forebrain of GAERS at postnatal days (PN)14, PN20, and PN90 and in age matched non-epileptic control Wistar rats. In PN14 GAERS, when no SWDs have developed yet, IL-1beta immunostaining was undetectable, and astrocytes and microglia showed a resting phenotype similar to control Wistar rats. In 3 out of 9 PN20 GAERS, IL-1beta was observed in activated astrocytes of the somatosensory cortex; the cytokine expression was associated with the occurrence of immature type of SWDs. In all adult PN90 GAERS, when mature SWDs are established, IL-1beta was observed in reactive astrocytes of the somatosensory cortex but not in adjacent cortical areas or in extra-cortical regions. An age-dependent c-fos activation was found in the somatosensory cortex of GAERS with maximal levels reached in PN90 rats; c-fos was also induced in some thalamic nuclei in PN20 and PN90 GAERS. Inhibition of IL-1beta biosynthesis in PN90 GAERS by 4-day systemic administration of a specific ICE/Caspase-1 blocker, significantly reduced both SWD number and duration. These results show that IL-1beta is induced in reactive astrocytes of the somatosensory cortex of GAERS at the onset of SWDs. IL-1beta has pro-ictogenic properties in this model, and thus it may play a contributing role in the mechanisms underlying the occurrence of absence seizures. PMID- 21645620 TI - DJ-1 associates with synaptic membranes. AB - Parkinson's disease (PD) is a neurodegenerative disorder caused by loss of dopaminergic neurons. Although many reports have suggested that genetic factors are implicated in the pathogenesis of PD, molecular mechanisms underlying selective dopaminergic neuronal degeneration remain unknown. DJ-1 is a causative gene for autosomal recessive form of PARK7-linked early-onset PD. A number of studies have demonstrated that exogenous DJ-1 localizes within mitochondria and the cytosol, and functions as a molecular chaperone, as a transcriptional regulator, and as a cell protective factor against oxidative stress. However, the precise subcellular localization and function of endogenous DJ-1 are not well known. The mechanisms by which mutations in DJ-1 contributes to neuronal degeneration also remain poorly understood. Here we show by immunocytochemistry that DJ-1 distributes to the cytosol and membranous structures in a punctate appearance in cultured cells and in primary neurons obtained from mouse brain. Interestingly, DJ-1 colocalizes with the Golgi apparatus proteins GM130 and the synaptic vesicle proteins such as synaptophysin and Rab3A. Forster resonance energy transfer analysis revealed that a small portion of DJ-1 interacts with synaptophysin in living cells. Although the wild-type DJ-1 protein directly associates with membranes without an intermediary protein, the pathogenic L166P mutation of DJ-1 exhibits less binding to synaptic vesicles. These results indicate that DJ-1 associates with membranous organelles including synaptic membranes to exhibit its normal function. PMID- 21645621 TI - High levels of human recombinant cyclooxygenase-1 expression in mammalian cells using a novel gene amplification method. AB - We report the expression of a high level of human cyclooxygenase-1 (hCOX-1) in mammalian cells using a novel gene amplification method known as the IR/MAR gene amplification system. IR/MAR-plasmids contain a mammalian replication initiation region (IR) and a nuclear matrix attachment region (MAR) and amplify autonomously without a specific induction process. In this study, the IR/MAR-plasmid pDeltaBN.AR1 was cotransfected with pCAG-COX1, which expresses hCOX-1, into human HEK293T cells, and G418 and blasticidin S double-resistant cells were obtained in about 1month. Real-time PCR and Western blotting revealed that the expressions of hCOX-1 mRNA and protein in both polyclonal and monoclonal cells were remarkably higher than those in only pCAG-COX1-transfected control cells. Southern blotting demonstrated the amplification of the hCOX-1 gene, and the copy number of clone #43 obtained by the cotransfection of pDeltaBN.AR1 and pCAG-COX1 was more than 20 copies per cell, though that of clone #14 obtained without using the IR/MAR plasmid pDeltaBN.AR1 was only two copies. These results indicate that a high level of hCOX-1 expression was achieved as a result of hCOX-1 gene amplification. Furthermore, the crude extract from clone #43 showed a strong COX-1 activity, and the activity was inhibited by the representative COX-1 inhibitor indomethacin, with an IC(50) value of 36nM. These results demonstrate that the IR/MAR gene amplification system is a simple but useful method for generating highly productive mammalian cells. PMID- 21645622 TI - Francisella noatunensis in Atlantic cod (Gadus morhua L.); waterborne transmission and immune responses. AB - This is the first report that confirms waterborne transmission of francisellosis in Atlantic cod. To investigate the transmission of disease, particle reduced water was transferred from a tank with intraperitoneally infected cod to a tank with healthy cod. Waterborne transmission of Francisella noatunensis was confirmed in the effluent group using immunohistochemistry and real-time quantitative PCR (RT-qPCR). The bacteria were located inside the accumulated macrophage-like cells. Specific and high antibody responses against live and inactivated bacteria were observed. Oil adjuvant had no effect on the antibody responses against inactivated F. noatunensis compared to saline formulation. The antigen epitope was a 20-25 kDa component of F. noatunensis suggested to be lipopolysaccharide detected by Western blot, Sypro Ruby and Silver staining. Systemic immune reactions were investigated by measuring the expression of IFN gamma, IL-1beta and IL-10 genes with RT-qPCR. After i.p. injection of live bacteria, a significant up-regulation of IFN-gamma and IL-1beta expression was observed from 15 to 60 days post infection in spleen and head kidney. In intestine, IFN-gamma was significantly up-regulated after 30 days whereas rectum showed no significant differences in expression. Elevated expression of IL-10 was observed in all the organs tested but was only significantly up-regulated at 60 days post infection in intestine from i.p. infected fish. For the cohabitant group, IL-1beta and IFN-gamma was up-regulated in spleen whereas intestine and rectum showed a down-regulation after 60 days. IL-10 was up-regulated in intestine of cohabitant fish from day 30 to day 60. These results indicate that F. noatunensis infection provokes both specific antibody responses and long term inflammatory responses in cod. The present study provides new knowledge about infection routes and shows that both humoral and cellular defence mechanisms are triggered by F. noatunensis in cod. PMID- 21645623 TI - Granger causality with signal-dependent noise. AB - It is generally believed that the noise variance in in vivo neuronal data exhibits time-varying volatility, particularly signal-dependent noise. Despite a widely used and powerful tool to detect causal influences in various data sources, Granger causality has not been well tailored for time-varying volatility models. In this technical note, a unified treatment of the causal influences in both mean and variance is naturally proposed on models with signal-dependent noise in both time and frequency domains. The approach is first systematically validated on toy models, and then applied to the physiological data collected from Parkinson patients, where a clear advantage over the classical Granger causality is demonstrated. PMID- 21645624 TI - Induced gamma band responses in human EEG after the control of miniature saccadic artifacts. AB - Induced gamma band responses (iGBRs) in the human electroencephalogram (EEG) have been ascribed to the activation of cortical object representations. Recently, this claim was challenged and it was stated that iGBRs occurring in the time window between 200 and 350 ms after stimulus onset are, to a great extent, generated by an electromyogenic artifact caused by miniature saccades (MS). In the present paper we focus on the characterization of iGBRs during the activation of cortical object representations, when recordings have been controlled for saccade-related transient potentials. For this we present an algorithm for the correction of saccade-related transient potentials (COSTRAP) which identifies and notably suppresses transient spike potentials (TSPs) that are likely to be linked to MSs. Furthermore, we conducted an EEG study to demonstrate (1) the feasibility of the algorithm, (2) the cortical origin iGBRs and (3) their relation to cortical object representations. Our results revealed that (i) it is possible to isolate TSPs, (ii) the morphology of the cleansed iGBR cannot be explained by an underlying myogenic artifact and (iii) the remaining iGBRs are sensitive to object recognition. Therefore we conclude that, with saccadic artifacts being controlled, high-frequency oscillations in human EEG are reliable electrophysiological correlates of cognitive processes. PMID- 21645625 TI - Dissociable systems of working memory for rhythm and melody. AB - Specialized neural systems are engaged by the rhythmic and melodic components of music. Here, we used PET to measure regional cerebral blood flow (rCBF) in a working memory task for sequences of rhythms and melodies, which were presented in separate blocks. Healthy subjects, without musical training, judged whether a target rhythm or melody was identical to a series of subsequently presented rhythms or melodies. When contrasted with passive listening to rhythms, working memory for rhythm activated the cerebellar hemispheres and vermis, right anterior insular cortex, and left anterior cingulate gyrus. These areas were not activated in a contrast between passive listening to rhythms and a non-auditory control, indicating their role in the temporal processing that was specific to working memory for rhythm. The contrast between working memory for melody and passive listening to melodies activated mainly a right-hemisphere network of frontal, parietal, and temporal cortices: areas involved in pitch processing and auditory working memory. Overall, these results demonstrate that rhythm and melody have unique neural signatures not only in the early stages of auditory processing, but also at the higher cognitive level of working memory. PMID- 21645626 TI - Phylogeographic structure and demographic patterns of brown trout in North-West Africa. AB - The objectives of the study were to determine the phylogeographic structure of brown trout (Salmo trutta) in Morocco, elucidate their colonization patterns in North-West Africa and identify the mtDNA lineages involved in this process. We also aimed to resolve whether certain brown trout entities are also genetically distinct. Sixty-two brown trout from eleven locations across the Mediterranean and the Atlantic drainages in Morocco were surveyed using sequence analysis of the mtDNA control region and nuclear gene LDH, and by genotyping twelve microsatellite loci. Our study confirms that in Morocco both the Atlantic and Mediterranean basins are populated by Atlantic mtDNA lineage brown trout only, demonstrating that the Atlantic lineage (especially its southern clade) invaded initially not only the western part of the Mediterranean basin in Morocco but also expanded deep into the central area. Atlantic haplotypes identified here sort into three distinct groups suggesting Morocco was colonized in at least three successive waves (1.2, 0.4 and 0.2-0.1 MY ago). This notion becomes more pronounced with the finding of a distinct haplotype in the Dades river system, whose origin appears to coalesce with the nascent stage of the basal mtDNA evolutionary lineages of brown trout. According to our results, Salmo akairos, Salmo pellegrini and "green trout" from Lake Isli do not exhibited any character states that distinctively separate them from the other brown trout populations studied. Therefore, their status as distinct species was not confirmed. PMID- 21645627 TI - Evolution of semi-quantitative whole joint assessment of knee OA: MOAKS (MRI Osteoarthritis Knee Score). AB - OBJECTIVE: In an effort to evolve semi-quantitative scoring methods based upon limitations identified in existing tools, integrating expert readers' experience with all available scoring tools and the published data comparing the different scoring systems, we iteratively developed the magnetic resonance imaging (MRI) Osteoarthritis Knee Score (MOAKS). The purpose of this report is to describe the instrument and its reliability. METHODS: The MOAKS instrument refines the scoring of bone marrow lesions (BMLs) (providing regional delineation and scoring across regions), cartilage (sub-regional assessment), and refines the elements of meniscal morphology (adding meniscal hypertrophy, partial maceration and progressive partial maceration) scoring. After a training and calibration session two expert readers read MRIs of 20 knees separately. In addition, one reader re read the same 20 MRIs 4 weeks later presented in random order to assess intra rater reliability. The analyses presented here are for both intra- and inter rater reliability (calculated using the linear weighted kappa and overall percent agreement). RESULTS: With the exception of inter-rater reliability for tibial cartilage area (kappa=0.36) and tibial osteophytes (kappa=0.49); and intra-rater reliability for tibial BML number of lesions (kappa=0.54), Hoffa-synovitis (kappa=0.42) all measures of reliability using kappa statistics were very good (0.61-0.8) or reached near-perfect agreement (0.81-1.0). Only intra-rater reliability for Hoffa-synovitis, and inter-rater reliability for tibial and patellar osteophytes showed overall percent agreement <75%. CONCLUSION: MOAKS scoring shows very good to excellent reliability for the large majority of features assessed. Further iterative development and research will include assessment of its validation and responsiveness. PMID- 21645628 TI - Nitrones are able to release nitric oxide in aqueous environment under hydroxyl free radical attack. AB - Importance of a nitric oxide donor that can act as a spin trap might bring some new therapeutic possibilities regarding the treatment of ischemic diseases by reducing the intensity of free radical produced reperfusion lesions. These substances might be also used as a new type of photo protectors since they can absorb UV radiation, capture free radicals formed by interaction of UV radiation with tissue constituents, and tanning of the skin will be permitted due to nitric oxide release. The purpose of this work was to measure the ability of nitrones to release nitric oxide and how different factors (temperature, nitrone concentration, and free radicals) influence the releasing ability. Mostly, indirect determination of nitric oxide was carried out, by measuring nitrite and nitrate amounts (as decomposition products of nitric oxide), all nitrones proved to release significant amounts of nitric oxide. Nitrite measurements were made based on an HPLC-VIS method that uses pre-column derivatization of nitrite by forming an azo dye (limit of quantification: 5ng/ml). No good correlation was found between the amount of nitric oxide and temperature for most studied nitrones but between the formation of nitric oxide and nitrone concentration an asymptotic correlation was found. Fenton reagent also yielded formation of nitric oxide from nitrones and formed amounts were not different from those recorded for UV irradiation. Most of the nitrones effectively released about 0.5% of the maximum amount of nitric oxide that is chemically possible and estimated concentrations of 0.1MUM were present in the solutions during decomposition. PMID- 21645629 TI - Extracellular phospholipase activity of Malassezia strains isolated from individuals with and without dermatological disease. AB - BACKGROUND: The Malassezia genus includes mainly lipophilic yeasts belonging to the cutaneous microbiota of man and other mammals. Some Malassezia species have been associated with various dermatological diseases. The factors permitting the transformation of yeasts of the Malassezia genus from a commensal organism to a pathogenic agent are still little known but the production of various enzymes such as lipase, phospholipase and lipoxygenase could contribute to the pathogenic activity of these yeasts. AIMS: Here we have determined and compared the extracellular phospholipase activity of sixty human isolates of Malassezia so as to relate this feature to the species of Malassezia and to the origin (from dermatological diseases or not) of the strains examined. METHODS: Phospholipase production was determined using the semi-quantitative egg-yolk plate method. RESULTS AND CONCLUSIONS: Malassezia obtusa, Malassezia slooffiae, Malassezia globosa, Malassezia restricta had difficulty developing in the chosen culture medium so that it was not possible to measure phospholipasic activity. Malassezia pachydermatis showed the highest phospholipase activity. Twenty-nine Malassezia sympodialis strains produced phospholipase; the isolates from patients with pityriasis versicolor had significantly higher phospholipasic activity than those isolated from healthy individuals. This observation suggests that the phospholipasic activity of Malassezia may play a role in the onset of skin lesions, especially in the case of pityriasis versicolor. PMID- 21645630 TI - Towards a more reliable comet assay: optimising agarose concentration, unwinding time and electrophoresis conditions. AB - The comet assay is now the method of choice for measuring most kinds of DNA damage in cells. However, due to the lack of a standardised protocol inter laboratory comparisons are of limited value. The aim of this paper is to demonstrate how small changes in comet-assay variables may significantly affect the results. We examined the effect of varying agarose concentrations, alkaline unwinding time, electrophoresis time, voltage and current, by use of two cell types, viz. human peripheral blood lymphocytes and the lymphoblastoid cell line TK-6. All these variables have marked effects on assay performance and, therefore, on the determination of DNA damage. Here we identify factors of particular importance. PMID- 21645631 TI - Genotoxicity testing of two lead-compounds in somatic cells of Drosophila melanogaster. AB - The in vivo genotoxic activity of two inorganic lead compounds was studied in Drosophila melanogaster by measurement of two different genetic endpoints. We used the wing-spot test and the comet assay. The comet assay was conducted with larval haemocytes. The results from the wing-spot test showed that neither lead chloride, PbCl(2), nor lead nitrate, Pb(NO(3))(2), were able to induce significant increases in the frequency of mutant spots. In addition, the combined treatments with gamma-radiation and PbCl(2) or Pb(NO(3))(2) did not show significant variations in the frequency of the three categories of mutant spots recorded, compared with the frequency induced by gamma-radiation alone. This seems to indicate that the lead compounds tested do not interact with the repair of the genetic damage induced by ionizing radiation. When the lead compounds were evaluated in the in vivo comet assay with haemocytes, Pb(NO(3))(2) was effective in inducing significant increases of DNA damage with a direct dose-response pattern. These results confirm the usefulness of the comet assay with haemocytes as an in vivo model and support the assumption that there is a genotoxic risk associated with lead exposure. PMID- 21645632 TI - Boronic acids-a novel class of bacterial mutagen. AB - Boronic acids and their esters are important building blocks in organic syntheses including those for drug substances and for which, as far as it can be determined, there are no published reports of testing for genotoxicity. A number of boronic acids have now been tested in this laboratory using Salmonella typhimurium strains TA1535, TA1537, TA98 and TA100 and Escherichia coli strain WP2uvrA(pKM101). Twelve of the 13 structures presented here were found to be mutagenic. All the compounds except one were active only in TA100 and/or WP2uvrA(pKM101), did not require S9 activation and produced relatively weak responses, i.e. no more than seven times the concurrent solvent-control values at >1000MUg/plate. The single exception was also weakly mutagenic for TA1537 in the presence of S9. Results with two compounds mutagenic for both TA100 and WP2uvrA(pKM101) showed no evidence of DNA-adduct formation detectable by (32)P postlabelling. It appears that boronic acids represent a novel class of bacterial mutagen that may not act by direct covalent binding to DNA. However, their mechanism of action remains to be elucidated and it cannot yet be determined whether or not they present a real genotoxic hazard. PMID- 21645633 TI - Chromosomal instability at the 7q11.23 region impacts on DNA-damage response in lymphocytes from Williams-Beuren syndrome patients. AB - Williams-Beuren syndrome (WBS) is the chromosomal disorder arising from a hemizygous microdeletion at 7q11.23. The present study was focused on a comparative investigation of genomic integrity in WBS patients by use of cytogenetic methods and the alkaline comet assay. Lymphocytes of whole peripheral blood were cultured and metaphases were examined for frequency and spectrum of chromosome aberrations. A WBS-related microdeletion was detected by means of the FISH (fluorescence in situ hybridization) technique. The blood samples from patients who were carriers of this microdeletion, were tested in the comet assay. For this purpose, freshly collected lymphocytes were exposed to hydrogen peroxide (100MUM, 1min, 4 degrees C). The frequencies of endogenous and exogenous DNA damage, and the kinetics and efficiency of DNA repair were measured during three subsequent hours of incubation. Comparison of the two data sets in this group of patients demonstrated a slightly elevated average frequency of chromosome aberrations, significantly increased levels of endogenous and H(2)O(2)-induced DNA damage, and somewhat impaired DNA repair. The relationship between an abnormal DNA-damage response and the 7q11.23 hemizygous microdeletion was confirmed experimentally when comparing the comet assay data in FISH-positive and FISH-negative lymphocytes from WBS-suspected patients. Briefly, our results indicate the impact of chromosomal instability within this region on susceptibility towards DNA damage, which may contribute to pathogenesis of this disease. It was shown also that the comet assay, as well as an experimental design proposed here, seem to be useful tools for estimating genome integrity in WBS patients. PMID- 21645634 TI - Efficacy of chloroquine plus primaquine treatment and pfcrt mutation in uncomplicated falciparum malaria patients in Rangamati, Bangladesh. AB - A combination of chloroquine (CQ) and primaquine (PQ) had been used as the first line treatment of uncomplicated Plasmodium falciparum malaria in Rangamati, Bangladesh until the end of 2004. Doctors or medical staffs had felt that CQ plus PQ had become less effective against uncomplicated falciparum malaria patients, but that it was more effective against the minority-indigenous patients than the Bengali patients. The efficacy of CQ plus PQ and the mutation status of the CQ resistance transporter (pfcrt) gene of infecting P. falciparum were, thus, investigated for 45 uncomplicated falciparum malaria patients in Rangamati in 2004. The total failure rate was 57.8%. One or two pfcrt sequences (CIETH and SMNTH at positions 72, 74-76, and 97, mutation underlined) with K76T mutation known to be related to CQ-resistant phenotype were detected in 38 patients' blood samples. Of the 38 patients, in total 15 patients (14/25 minority-indigenous and 1/13 Bengali patients) resulted in adequate clinical and parasitological response (ACPR). There was a statistically significant difference in ACPR rate between the minority-indigenous patients and the Bengali patients. P. falciparum with mutant or resistant pfcrt (pfcrt-resistant) was detected by PCR in blood samples on day 28 for 10 ACPR minority-indigenous patients but not for the only one Bengali ACPR patient, who all were infected with pfcrt-resistant P. falciparum on day 0. The minority-indigenous patients, but not Bengalis, are suggested to be often cured by CQ plus PQ, leaving a very few parasites detectable only by PCR, even when they are infected with pfcrt-resistant P. falciparum. PMID- 21645636 TI - Participatory design for computerization of clinical practice guidelines. AB - There have been made many attempts on computerization of clinical practice guidelines (CPGs), none have, however achieved any general application in clinical work practice. The objective of this paper is: (1) to raise awareness about the impact the design method used for computerization of CPGs have on the final solution and (2) to explore the potential benefits--and disadvantages--of participatory design (PD) as an approach to design. However, rather than attempting to comprehensively cover the whole field of PD pertinent to healthcare, we focus on providing details on three aspects of PD: PD as a design philosophy, PD as a toolbox and PD as a way to create a shared realm of understanding among IT-designers and health professionals as these are areas of utmost relevance for the design of computerized CPGs. Additionally, the application of PD for computerization of CPGs is illustrated by two cases. We conclude that PD is a beneficial approach for design of computerized CPGs. PMID- 21645635 TI - Desorption electrospray ionization mass spectrometry for lipid characterization and biological tissue imaging. AB - Desorption electrospray ionization mass spectrometry (DESI-MS) imaging of biological samples allows untargeted analysis and structural characterization of lipids ionized from the near-surface region of a sample under ambient conditions. DESI is a powerful and sensitive MS ionization method for 2D and 3D imaging of lipids from direct and unmodified complex biological samples. This review describes the strengths and limitations of DESI-MS for lipid characterization and imaging together with the technical workflow and a survey of applications. Included are discussions of lipid mapping and biomarker discovery as well as a perspective on the future of DESI imaging. PMID- 21645637 TI - Clinical data interoperability based on archetype transformation. AB - The semantic interoperability between health information systems is a major challenge to improve the quality of clinical practice and patient safety. In recent years many projects have faced this problem and provided solutions based on specific standards and technologies in order to satisfy the needs of a particular scenario. Most of such solutions cannot be easily adapted to new scenarios, thus more global solutions are needed. In this work, we have focused on the semantic interoperability of electronic healthcare records standards based on the dual model architecture and we have developed a solution that has been applied to ISO 13606 and openEHR. The technological infrastructure combines reference models, archetypes and ontologies, with the support of Model-driven Engineering techniques. For this purpose, the interoperability infrastructure developed in previous work by our group has been reused and extended to cover the requirements of data transformation. PMID- 21645638 TI - Automatic figure classification in bioscience literature. AB - Millions of figures appear in biomedical articles, and it is important to develop an intelligent figure search engine to return relevant figures based on user entries. In this study we report a figure classifier that automatically classifies biomedical figures into five predefined figure types: Gel-image, Image of-thing, Graph, Model, and Mix. The classifier explored rich image features and integrated them with text features. We performed feature selection and explored different classification models, including a rule-based figure classifier, a supervised machine-learning classifier, and a multi-model classifier, the latter of which integrated the first two classifiers. Our results show that feature selection improved figure classification and the novel image features we explored were the best among image features that we have examined. Our results also show that integrating text and image features achieved better performance than using either of them individually. The best system is a multi-model classifier which combines the rule-based hierarchical classifier and a support vector machine (SVM) based classifier, achieving a 76.7% F1-score for five-type classification. We demonstrated our system at http://figureclassification.askhermes.org/. PMID- 21645639 TI - Lactated Ringer's solution reduces systemic inflammation compared with saline in patients with acute pancreatitis. AB - BACKGROUND & AIMS: Aggressive fluid resuscitation is recommended for initial management of acute pancreatitis. We performed a randomized controlled trial to evaluate the impact of a goal-directed fluid resuscitation protocol on systemic inflammation in patients with acute pancreatitis. We then determined the impact of resuscitation with lactated Ringer's solution, compared with normal saline. METHODS: We performed a randomized controlled trial of 40 patients with acute pancreatitis at 3 New England hospitals from May 2009-February 2010. Patients received goal-directed fluid resuscitation with lactated Ringer's solution, goal directed fluid resuscitation with normal saline, standard fluid resuscitation with lactated Ringer's solution, or standard fluid resuscitation with normal saline. Systemic inflammation was measured on the basis of levels of systemic inflammatory response syndrome (SIRS) and C-reactive protein (CRP) level after 24 hours. RESULTS: The volumes of fluid administered during a 24-hour period were similar among patients given goal-directed or standard fluid resuscitation (mean, 4300 vs 4600 mL, respectively; P = .87). Goal-directed resuscitation did not significantly reduce incidence of SIRS, compared with standard resuscitation (11.8% vs 13.0%, respectively; P = .85) or levels of CRP after 24 hours (87.1 vs 69.2 mg/dL, respectively; P = .75). By contrast, there was a significant reduction in SIRS after 24 hours among subjects resuscitated with lactated Ringer's solution, compared with normal saline (84% reduction vs 0%, respectively; P = .035); administration of lactated Ringer's solution also reduced levels of CRP, compared with normal saline (51.5 vs 104 mg/dL, respectively; P = .02). CONCLUSIONS: Patients with acute pancreatitis who were resuscitated with lactated Ringer's solution had reduced systemic inflammation compared with those who received saline. PMID- 21645640 TI - Common symptoms and stressors among individuals with inflammatory bowel diseases. AB - BACKGROUND & AIMS: We evaluated symptoms and stressful life events over a 1-year period in a population-based sample of persons with inflammatory bowel disease (IBD). METHODS: Participants from the University of Manitoba IBD Research Registry (n = 704) completed 5 surveys, given every 3 months for 1 year (552 completed all the surveys). Respondents were asked to indicate the specific gastrointestinal and other symptoms, if any, they had experienced in the previous 3-month period and to document any significant stressors experienced. The Manitoba IBD Index was used to categorize active versus inactive disease. RESULTS: In any 3-month period, participants with Crohn's disease, compared with those with ulcerative colitis (UC), reported more diarrhea (63% vs 38%), fatigue (54% vs 33%), abdominal pain (47% vs 32%), aching joints (42% vs 29%), painful joints (24% vs 16%), fever or night sweats (24% vs 15%), nausea/vomiting (18% vs 7%), and reductions in appetite (19% vs 11%) (P < .001 for each symptom). Individuals with ulcerative colitis complained more of stool mucous or blood than those with Crohn's disease (27% vs 17%; P < .001). In periods of inactive disease, participants still experienced symptoms such as aching joints (17%), fatigue (15%), diarrhea (13%), or abdominal pain (9%). In any 3-month period, approximately 50% experienced some type of stress; family stress was the most commonly reported form, followed by work or school and financial stress. CONCLUSIONS: Diarrhea and fatigue are the 2 most common symptoms of individuals with IBD. Those with inactive disease still report symptoms. Almost 50% of participants reported significant stress in any 3-month period, but the primary types were everyday life stressors more so than health-related stress. PMID- 21645641 TI - Targeting properties of peptide-modified radiolabeled liposomal nanoparticles. AB - Radiolabeled PEGylated liposomal nanoparticles (NPs) open new possibilities for a variety of applications including diagnosis, drug delivery, targeted therapy, and monitoring treatment effects. Here we describe the characterization of liposomal NPs (liposomes and micelles) derivatized with the somatostatin analogue tyrosine 3-octreotide as a proof of concept for tumor targeting. NPs were radiolabeled with indium-111, and targeting properties were evaluated in vitro on rat pancreatic tumor cells (AR42J), demonstrating specific binding and IC(50) values in the low nanomolar range. Biodistribution studies were performed in Lewis rats and compared to single-photon emission computed tomography images. Moderate tumor uptake was found in xenografted nude mice (<2.5% ID/g tissue) as compared to control. Micelles and liposomes revealed comparable pharmacokinetics and targeting properties. This study provides insight into tumor-targeting characteristics of peptide-derivatized liposomal NPs and can serve as a basis for further improvement of these constructs. FROM THE CLINICAL EDITOR: The authors investigated tumor-targeting characteristics of peptide-derivatized liposomal NPs. Similar radiolabeled PEGylated liposomal NPs open new possibilities for a variety of applications including diagnosis, drug delivery, targeted therapy, and treatment monitoring. PMID- 21645642 TI - A comprehensive biological evaluation of ceramic nanoparticles as wear debris. AB - Patients are exposed internally to nanoparticles (NPs) by wear mechanisms associated with total joint arthroplasty. This tissue-specific retention implies that the biological evaluation of NPs shall be integrative and niche targeting. Here, we report that ceramic zirconia and silicon nitride NPs interfere with MG63 cells' function and remarkably stimulate the secretion of TNF-alpha in RAW264.7 cells. However, alumina NPs significantly promote the alkaline phosphatase (ALP) activity of MG63 cells at low concentration and do not show irritation to macrophages. In this study, we prove that ceramic materials at nanoscale are bioactive to cells. These findings also suggest that a more rational paradigm for the biosafety evaluation of NPs than is currently in place is needed before their clinical applications. FROM THE CLINICAL EDITOR: In this study, the authors demonstrate that ceramic nano materials associated with normal wear-and-tear of joint prostheses at nanoscale are bioactive to cells. PMID- 21645643 TI - Intraperitoneal photodynamic therapy mediated by a fullerene in a mouse model of abdominal dissemination of colon adenocarcinoma. AB - Functionalized fullerenes represent a new class of photosensitizer (PS) that is being investigated for photodynamic therapy (PDT) of various diseases, including cancer. We tested the hypothesis that fullerenes could be used to mediate PDT of intraperitoneal (IP) carcinomatosis in a mouse model. In humans this form of cancer responds poorly to standard treatment and manifests as a thin covering of tumor nodules on intestines and on other abdominal organs. We used a colon adenocarcinoma cell line (CT26) stably expressing luciferase to allow monitoring of IP tumor burden in BALB/c mice by noninvasive real-time optical imaging using a sensitive low-light camera. IP injection of a preparation of N methylpyrrolidinium-fullerene formulated in Cremophor-EL micelles, followed by white-light illumination delivered through the peritoneal wall (after creation of a skin flap), produced a statistically significant reduction in bioluminescence and a survival advantage in mice. FROM THE CLINICAL EDITOR: This team of investigators report on functionalized fullerenes, to be used as photosensitizer for photodynamic therapy and demonstrate the efficacy of this method in an intraperitoneal carcinomatosis mouse model. PMID- 21645644 TI - A hybrid approach to predicting events in clinical trials with time-to-event outcomes. AB - In many clinical trials with time-to-event outcomes there are interim analyses planned at pre-specified event counts. It is of great value to predict when the pre-specified event milestones can be reached based on the available data as the timeline for a study is essential to the study sponsors and data monitoring committees for logistic planning purposes. Both parametric and non-parametric approaches exist in the literature for estimating the underlining survival function, based on which the predictions of future event times can be determined. The parametric approaches assume that the survival function is smooth, which is often not the case as the survival function usually has one or multiple change points and the hazard functions can differ significantly before and after a change point. The existing non-parametric method bases predictions on the Kaplan Meier survival curve appended with a parametric tail to the largest observation, and all of the available data is used to estimate the parametric tail. This approach also requires a smooth survival function in order to achieve an accurate estimate of the tail distribution. In this article, we propose a hybrid parametric, non-parametric approach to predicting events in clinical trials with time-to-event outcomes. The change points in the survival function are first detected, and the survival function before the last change point is estimated non parametrically and the tail distribution beyond the last change point is estimated parametrically. Numerical results show that the proposed approach provides accurate predictions for future event times and outperforms the existing approaches. PMID- 21645645 TI - Coding and non-coding polymorphisms in VDR gene and susceptibility to pulmonary tuberculosis in tribes, castes and Muslims of Central India. AB - Vitamin D receptor (VDR) plays an important role in activating immune response against various infectious agents. This study was aimed to investigate the association between VDR gene polymorphisms and different clinical forms of pulmonary tuberculosis (TB) in different population groups. Four common polymorphisms (TaqI, ApaI, BsmI and FokI) of VDR gene were studied in clinically diagnosed TB patients and healthy controls from Sahariya tribe (n=377), Bhil tribe (n=95), Chhattisgarh tribe (n=33), general population from North-Central (NC) (n=1021) and South-Eastern (SE) region (n=646) and Muslims (n=217). Genotyping was carried out using PCR-RFLP method and re-confirmed by direct sequencing. The haplotype analysis was performed on Haploview 4.1 and statistical analysis was done using SPSS 13.0 software. We found that bb genotype of BsmI polymorphism conferred significant risk to smear positive and multiple drug resistant (MDR) TB in tribes [OR (CI)=3.7 (1.5-9.2), p=0.002], SE population [OR (CI)=2.1 (1.4-3.3), p=0.0004] and Muslims [OR (CI)=6.7 (1.1-39), p=0.01]. The subjects with FF genotype of FokI polymorphism appeared less likely (p=0.004) to develop MDR TB in NC population, whereas, those with Ff [OR (CI)=2.5 (1.3-5.0), p=0.004] and ff [OR (CI)=3.4 (1.2-9.3), p=0.01] genotypes were at high risk of MDR and smear positive disease, respectively. Similarly, tt genotype of TaqI polymorphism was found associated with high risk of smear positive TB in NC [OR (CI)=3.6 (0.9-14.2), p=0.05] as well as in SE [OR (CI)=4.7 (1.8-12.3), p=0.00003] population. Interestingly, tt genotype appeared strongly associated [OR (CI)=8.9 (2.7-29), p=0.00001] with high bacillary load outcome. In conclusion, genetic polymorphisms in VDR gene, alone or in combination (haplotypes) are associated with different clinical outcomes in pulmonary TB. PMID- 21645646 TI - High genetic diversity of HIV-1 was found in men who have sex with men in Shijiazhuang, China. AB - Men who have sex with men (MSM) have become one of the populations with severely HIV prevalence in China. However, very few genetic studies have been done on HIV 1 spreading in this population. In this study, the genetic characterization of HIV-1 strains prevalent in the MSM in Shijiazhuang, China, was analyzed basing on the HIV-1 full-length gag, pol, and partial env gene. 21 drug-naive HIV-1 sero positive patients were enrolled into the study. Full length gag, pol, partial env genes and some near full length genomes were amplified with nest RT-PCR followed by sequencing. Multiple subtypes, including CRF01_AE (52.9%), subtype B (35.3%) and CRF07_BC (11.8%), were found in the population. Phylogenetic analysis showed close relationship between our strains with those from Beijing MSM but not those from paid blood donors of local area. Drug resistance surveillance evaluation showed no transmitted drug resistance (TDR). However, some drug-resistance related polymorphism sites (V181I and V106I) were popularly found in B subtype strains. The findings will contribute to understanding the distribution and evolution of HIV-1 in MSM and also help the behavior change intervention. PMID- 21645647 TI - Dynamics of hepatitis D (delta) virus genotype 3 in the Amazon region of South America. AB - Hepatitis delta virus (HDV) is widely distributed and associated with fulminant hepatitis epidemics in areas with high prevalence of HBV. Several studies performed in the 1980s showed data on HDV infection in South America, but there are no studies on the viral dynamics of this virus. The aim of this study was to conduct an evolutionary analysis of hepatitis delta genotype 3 (HDV/3) prevalent in South America: estimate its nucleotide substitution rate, determine the time of most recent ancestor (TMRCA) and characterize the epidemic history and evolutionary dynamics. Furthermore, we characterized the presence of HBV/HDV infection in seven samples collected from patients who died due to fulminant hepatitis from Amazon region in Colombia and included them in the evolutionary analysis. This is the first study reporting HBV and HDV sequences from the Amazon region of Colombia. Of the seven Colombian patients, five were positive for HBV DNA and HDV-RNA. Of them, two samples were successfully sequenced for HBV (subgenotypes F3 and F1b) and the five samples HDV positive were classified as HDV/3. By using all HDV/3 available reference sequences with sampling dates (n=36), we estimated the HDV/3 substitution rate in 1.07 * 10(-3) substitutions per site per year (s/s/y), which resulted in a time to the most recent common ancestor (TMRCA) of 85 years. Also, it was determined that HDV/3 spread exponentially from early 1950s to the 1970s in South America. This work discusses for the first time the viral dynamics for the HDV/3 circulating in South America. We suggest that the measures implemented to control HBV transmission resulted in the control of HDV/3 spreading in South America, especially after the important raise in this infection associated with a huge mortality during the 1950s up to the 1970s. The differences found among HDV/3 and the other HDV genotypes concerning its diversity raises the hypothesis of a different origin and/or a different transmission route. PMID- 21645648 TI - Inherited mitochondrial variants are not a major cause of age-related hearing impairment in the European population. AB - Mitochondrial DNA (mtDNA) mutations have been implicated in various age-related diseases. To further clarify the role of mtDNA variants in age-related hearing impairment (ARHI), we determined the DNA sequence of the entire mitochondrial genome of 400 individuals using the Affymetrix Human Mitochondrial Resequencing Array. These were the 200 worst hearing and the 200 best hearing from a collection of 947 Belgian samples. We performed association tests with individual mitochondrial variants, comparison of the mutation load, and association with European haplogroups and their interaction with environmental risk factors. We also tested the influence of rare variants on ARHI. None of these tests showed any association with ARHI. PMID- 21645649 TI - In vitro interaction between the ammonium transport protein AmtB and partially uridylylated forms of the P(II) protein GlnZ. AB - The ammonium transport family Amt/Rh comprises ubiquitous integral membrane proteins that facilitate ammonium movement across biological membranes. Besides their role in transport, Amt proteins also play a role in sensing the levels of ammonium in the environment, a process that depends on complex formation with cytosolic proteins of the P(II) family. Trimeric P(II) proteins from a variety of organisms undergo a cycle of reversible posttranslational modification according to the prevailing nitrogen supply. In proteobacteria, P(II) proteins are subjected to reversible uridylylation of each monomer. In this study we used the purified proteins from Azospirillum brasilense to analyze the effect of P(II) uridylylation on the protein's ability to engage complex formation with AmtB in vitro. Our results show that partially uridylylated P(II) trimers can interact with AmtB in vitro, the implication of this finding in the regulation of nitrogen metabolism is discussed. We also report an improved expression and purification protocol for the A. brasilense AmtB protein that might be applicable to AmtB proteins from other organisms. PMID- 21645650 TI - Poppea's bath liquor: the secret proteome of she-donkey's milk. AB - Donkey's milk is today categorized among the best mother's milk substitute for allergic newborns, due to its much reduced or absent allergenicity, coupled to excellent palatability and nutritional value. However, up to the present, only a handful of proteins had been characterized, just about the standard eight to ten major ones known in all types of milk. By exploiting the combinatorial peptide ligand library technology, and treating large volumes (up to 300 mL) of defatted, de-caseinized (whey) milk, we have been able to identify 106 unique gene products, by far the largest description so far of this precious nutrient. Due to poor knowledge of the donkey's genetic asset, only 10% of the proteins could be identified by consulting the data base of Equus asinus; the largest proportion (70%) could be identified by homology with the proteins of Equus caballus. PMID- 21645651 TI - LRP4 association to bone properties and fracture and interaction with genes in the Wnt- and BMP signaling pathways. AB - Osteoporosis is a common complex disorder in postmenopausal women leading to changes in the micro-architecture of bone and increased risk of fracture. Members of the low-density lipoprotein receptor-related protein (LRP) gene family regulates the development and physiology of bone through the Wnt/beta-catenin (Wnt) pathway that in turn cross-talks with the bone morphogenetic protein (BMP) pathway. In two cohorts of Swedish women: OPRA (n=1002; age 75 years) and PEAK-25 (n=1005; age 25 years), eleven single nucleotide polymorphisms (SNPs) from Wnt pathway genes (LRP4; LRP5; G protein-coupled receptor 177, GPR177) were analyzed for association with Bone Mineral Density (BMD), rate of bone loss, hip geometry, quantitative ultrasound and fracture. Additionally, interaction of LRP4 with LRP5, GPR177 and BMP2 were analyzed. LRP4 (rs6485702) was associated with higher total body (TB) and lumbar spine (LS) BMD in the PEAK-25 cohort (p=0.006 and 0.005 respectively), and interaction was observed with LRP5 (p=0.007) and BMP2 (p=0.004) for TB BMD. LRP4 also showed significant interaction with LRP5 for femoral neck (FN) and LS BMD in this cohort. In the OPRA cohort, LRP4 polymorphisms were associated with significantly lower fracture incidence overall (p=0.008-0.001) and fewer hip fractures (rs3816614, p=0.006). Significant interaction in the OPRA cohort was observed for LRP4 with BMP2 and GPR177 for FN BMD as well as for rate of bone loss at TB and FN (p=0.007-0.0001). In conclusion, LRP4 and interaction between LRP4 and genes in the Wnt and BMP signaling pathways modulate bone phenotypes including peak bone mass and fracture, the clinical endpoint of osteoporosis. PMID- 21645652 TI - Rapid monitoring of mono- and disaccharides in drinks, foodstuffs and foodstuff additives by capillary electrophoresis with contactless conductivity detection. AB - A capillary electrophoresis (CE) procedure with contactless conductivity detection (C(4)D) has been developed for monitoring of neutral mono- and disaccharides in drinks and foodstuffs. The separation of a mixture of seven neutral saccharides (glucose, fructose, galactose, mannose, ribose, sucrose and lactose) employed a quartz capillary, 5 MUm i.d., with an effective length of 18.3 cm, and 75 mM NaOH (pH 12.8) as the background electrolyte (BGE). The limit of detection (LOD) values obtained lied within a range from 0.4 MUmol L(-1) for lactose to 0.9 MUmol L(-1) for ribose, with a separation time shorter than 140 s. The procedure was successfully applied to determinations of saccharides in fruit juices, Coca-Cola, milk, red and white wines, yoghurts, honey and a foodstuff additive. PMID- 21645653 TI - Dynamic reaction cell ICP-MS for determination of total As, Cr, Se and V in complex matrices: still a challenge? A review. AB - Mass interferences, caused by atomic or polyatomic species and having the same mass/charge ratio of the analyte, can be a severe limit for a reliable assay of trace and ultratrace elements by ICP-MS. The DRCTM technology uses a reaction gas to overcome these interferences. Reactions of charge exchange, atom transfer, adduct formation, condensation and analyte association/condensation are the main mechanisms. Interfering ions tend to react with the gas exothermally, while, the analyte reacts endothermally. Selecting the most appropriate reaction gas in DRC ICP-MS is the very critical point for the determination of strongly interfered elements. A careful evaluation of the reaction mechanisms and the chemistry involved are required. The DRC allows the use of different gases, among them, ammonia (NH(3)), methane (CH(4)), hydrogen (H(2)) and oxygen (O(2)) are the most known, but there are other potentially useful gases like nitrous oxide (N(2)O), nitrogen oxide (NO), carbon dioxide (CO(2)), fluoromethane (CH(3)F), sulphur hexafluoride (SF(6)) and carbon disulfide (CS(2)). This paper provides a review on the analytical challenges for a reliable assay of As, Cr, Se and V by DRC-ICP MS and illustrates different approaches and mechanisms involved in the analysis of polymers, biological fluids (serum, urine and whole blood), rock, soil and particulate matter. PMID- 21645654 TI - Platinum-gold alloy nanoparticles and horseradish peroxidase functionalized nanocomposite as a trace label for ultrasensitive electrochemical detection of thrombin. AB - A novel tracer, platinum-gold alloy nanoparticles (Pt-AuNPs) and horseradish peroxidase (HRP) functionalized single-walled carbon nanotubes (SWCNTs) composite, is employed to label the secondary thrombin aptamer for constructing an ultrasensitive electrochemical aptasensor. Thionine, immobilized on functionalized SWCNTs, provides a pair of distinguished redox peak for electrochemical detection. Both the high-content Pt-AuNPs and HRP on SWCNTs amplify the electrochemical signal of thionine through electrocatalytic reduction of H(2)O(2). Differential pulse voltammetry (DPV) is employed to detect thrombin with different concentrations. The reduction peak current is logarithmically related to the concentration of thrombin in an extremely wide range from 10 fM to 5 nM with a detection limit of 3.6 fM. The dual signal amplification of Pt-AuNPs and HRP functionalized nanocomposite provides a promising way for ultrasensitive assay in electrochemical aptasensors. PMID- 21645655 TI - Comparing dissolved reactive phosphorus measured by DGT with ferrihydrite and titanium dioxide adsorbents: anionic interferences, adsorbent capacity and deployment time. AB - Two adsorbents (Metsorb and ferrihydrite) used in binding layers with the diffusive gradients in a thin film technique were evaluated for the measurement of dissolved reactive phosphorous (DRP) in synthetic and natural waters. Possible interferences were investigated with Cl(-) (up to 1.35 mol L(-1)) and SO(4)(2-) (up to 0.056 mol L(-1)) having no affect on either DGT binding layer, and HCO(3)( ) (up to 5.7 mmol L(-1)) having no effect on Metsorb-DGT, over 4 days. However, HCO(3)(-) interfered with the ferrihydrite-DGT measurement at concentrations typical of many natural waters (>=0.7 mmol L(-1)) after a deployment period of 1 2 days. The capacity of the Metsorb binding phase for DGT response was ~37,000 ng P, whereas the capacities of a low-mass (17.8 mg of adsorbent per DGT sampler) and high-mass (29.2mg of adsorbent per DGT sampler) ferrihydrite binding phase were substantially lower (~15,000 ng P and ~25,000 ng P, low-mass and high-mass, respectively). Increasing the capacity of the ferrihydrite adsorbent allowed the ferrihydrite-DGT to be utilized for up to 3 days before interference by HCO(3)(-) was observed. Seawater deployments demonstrated that even high-capacity ferrihydrite-DGT devices underestimated the DRP concentration by 37%, whereas Metsorb-DGT measurements were accurate. The Metsorb-DGT is superior to the ferrihydrite-DGT for determining DRP over deployment times greater than 1 day and in waters with >=0.7 mmol L(-1) HCO(3)(-). Based on the experience obtained from this detailed validation process, the authors propose a number of key requirements that need to be considered when developing new DGT binding layers, with testing the performance over longer deployment times being critical. PMID- 21645656 TI - Effective concentration difference model to study the effect of various factors on the effective diffusion coefficient in the dialysis membrane. AB - Cellulose acetate dialysis membrane (CDM) has been used in the diffusive gradients in thin films (DGT) technique, where accurate diffusion coefficients are essential for the assessment of the concentrations of labile metal in solution. Effective concentration difference model (ECDM), based on the assumption that the effective diffusion coefficient of metal ion in the dialysis membrane is determined by the effective concentration difference (DeltaC(e)) across the dialysis membrane, is proposed and applied to study the effect of ionic strength, binding agent, ligands and Donnan potential on the effective diffusion coefficient. The effective diffusion coefficients of Cd(2+) through the dialysis membrane immersed in receptor solutions with binding agent were almost the same as those in receptor solutions without binding agent at higher ionic strengths (0.01-1 M) but much higher than those at lower ionic strengths (0.001 0.0001 M). The effective diffusion coefficients of Cd(2+) through the dialysis membrane immersed in deionized water receptor solutions with binding agent were not significantly different from those in synthetic receptor solutions (receptor solutions with various ionic strengths) with binding agent. The DGT-labile fractions were measured in synthetic solutions and natural waters, which indicated that the effective diffusion coefficients, through the dialysis membrane immersed in the deionized water solution with binding agent as receptor solution and in the spiked natural water as source solution, were more suitable for DGT application. PMID- 21645657 TI - Comparison of surfactant-assisted shotgun methods using acid-labile surfactants and sodium dodecyl sulfate for membrane proteome analysis. AB - Three surfactant-assisted shotgun methods using acid labile surfactants, sodium-3 [(2-methyl-2-undecyl-1,3-dioxolan-4-yl)-methoxyl]-1-propanesulfonate (RapiGest) and 3-[3-(1,1-bisalkyloxyethyl)pyridin-1-yl]propane-1-sulfonate (PPS), and sodium dodecyl sulfate (SDS) were investigated for their applicability to membrane proteome analysis. It is shown that RapiGest is a preferred reagent for handling membrane proteomes of Escherichia coli and MCF7 cells for liquid chromatography tandem mass spectrometry (LC MS/MS) analysis of tryptic digests. The RapiGest method allowed identification of more peptides and proteins than the SDS and PPS methods and there was no apparent bias for the type of peptides and proteins identified by the RapiGest and SDS methods, while a slightly higher proportion of hydrophilic peptides and proteins were identified by the PPS method. The performance of the SDS and PPS methods is similar in terms of the numbers of peptides and proteins identified. Since the SDS method required the removal of SDS using a technique such as strong-cation exchange (SCX), we further investigated the effect of SCX on sample loss through analyzing the digest of an enriched E. coli membrane fraction as well as a standard protein, bovine serum albumin (BSA). The results showed that extensive sample loss (as much as 62%) was encountered during the SCX cleaning step. We then applied the RapiGest method in combination with two-dimensional LC MS/MS to characterize the E. coli membrane proteome. In total, 1626 unique proteins (5799 unique peptides) were identified with a peptide false discovery rate of 2.4%. About 60% of the identified proteins with known cellular locations were found to be membrane proteins. Among them, about 75% were integral membrane proteins. This work represents one of the most comprehensive profiles of E. coli membrane proteome generated by a proteomic technique. PMID- 21645658 TI - Cross-talk-free simultaneous fluoroimmunoassay of two biomarkers based on dual color quantum dots. AB - In this article, we demonstrate the fabrication and simultaneous fluorescent detection of two biomarkers related to lung cancer. Polystyrene microspheres (PSM) were introduced as biomolecular immobilizing carriers and a 96-well filter plate was used as the separation platform. The whole experiment could be effectively carried out in a homogeneous system, as exemplified by the detection of carcinoembryonic antigen (CEA) and neuron specific enolase (NSE). First, two capture antibodies for CEA and NSE were immobilized on the PSM surface. Next, they reacted successively with two antigens and two modified detection antibodies. Finally, these two biomarkers could be recognized by streptavidin conjugated quantum dots (QD) and goat-anti-FITC conjugated QD with a detection limit of 0.625 ng mL(-1), which was lower than the clinical cut-off level. The protocol showed good precision within 6.36% and good recovery in the range of 90.86-105.02%. Compared with several other assay formats reported previously, our new technique is competitive or even better. Furthermore, the immunosensor was successfully illustrated in 20 serum samples. Overall, this new immunoassay offers a promising alternative for the detection of biomarkers related to cancer diseases, taking advantage of simplicity, specificity, sensitivity and cost efficiency. PMID- 21645659 TI - Development and validation of a fast monoclonal based disequilibrium enzyme linked immunosorbent assay for the detection of triphenylmethane dyes and their metabolites in fish. AB - Malachite Green (MG), Crystal Violet (CV) and Brilliant Green (BG) are antibacterial, antifungal and antiparasitic agents that have been used for treatment and prevention of diseases in fish. These dyes are metabolized into reduced leuco forms (LMG, LCV, LBG) that can be present in fish muscles for a long period. Due to the carcinogenic properties they are banned for use in fish for human consumption in many countries including the European Union and the United States. HPLC and LC-MS techniques are generally used for the detection of these compounds and their metabolites in fish. This study presents the development of a fast enzyme-linked immunosorbent assay (ELISA) method as an alternative for screening purposes. A first monoclonal cell line producing antibodies to MG was generated using a hybridoma technique. The antibody had good cross-reactivates with related chromatic forms of triphenylmethane dyes such as CV, BG, Methyl Green, Methyl Violet and Victoria Blue R. The monoclonal antibody (mAb) was used to develop a fast (20 min) disequilibrium ELISA screening method for the detection of triphenylmethanes in fish. By introducing an oxidation step with 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) during sample extraction the assay was also used to detect the presence of the reduced metabolites of triphenylmethanes. The detection capability of the assay was 1 ng g(-1) for MG, LMG, CV, LCV and BG which was below the minimum required performance limit (MRPL) for the detection method of total MG (sum of MG and LMG) set by the Commission Decision 2004/25/EC (2 ng g(-1)). The mean recoveries for fish samples spiked at 0.5 MRPL and MRPL levels with MG and LMG were between 74.9 and 117.0% and inter- and intra-assay coefficients of variation between 4.7 and 25.7%. The validated method allows the analysis of a batch of 20 samples in two to three hours. Additionally, this procedure is substantially faster than other ELISA methods developed for MG/LMG thus far. The stable and efficient monoclonal cell line obtained is an unlimited source of sensitive and specific antibody to MG and other triphenylmethanes. PMID- 21645660 TI - Imprinted sol-gel electrochemical sensor for the determination of benzylpenicillin based on Fe3O4@SiO2/multi-walled carbon nanotubes-chitosans nanocomposite film modified carbon electrode. AB - Herein, a novel imprinted sol-gel electrochemical sensor based on multi-walled carbon nanotubes (MWNTs) doped with chitosan film on a carbon electrode has been developed. Prior to doped, the MWNTs have been decorated with Fe(3)O(4) nanoparticles which have been coated uniformly with SiO(2) layer. The characterization of imprinted sensor has been carried out by X-ray diffraction and scanning electron microscopy. The performance of the proposed imprinted sensor has been investigated using cyclic voltammetry and differential pulse voltammetry. The imprinted sensor offers a fast response and sensitive benzylpenicillin quantification. The fabricated benzylpenicillin imprinted sensor exhibits a linear response from 5.0*10(-8) to 1.0*10(-3) mol L(-1) with a detection limit of 1.5*10(-9) mol L(-1). For samples analysis, perfect recoveries of the imprinted sensor for benzylpenicillin indicated that the imprinted sensor was able to detect benzylpenicillin in real samples successfully. PMID- 21645661 TI - High temperature liquid chromatography hyphenated with ESI-MS and ICP-MS detection for the structural characterization and quantification of halogen containing drug metabolites. AB - In this paper we describe the hyphenation of high temperature liquid chromatography with ICP-MS and ESI-MS for the characterization of halogen containing drug metabolites. The use of temperature gradients up to 200 degrees C enabled the separation of metabolites with low organic modifier content. This specific property allowed the use of detection methods that suffer from (significant) changes in analyte response factors as a function of the organic modifier content such as ICP-MS. Metabolites of two kinase inhibitors (SB-203580 Iodo and MAPK inhibitor VIII) produced by bacterial cytochrome P450 BM3 mutants and human liver microsomes were identified based on high resolution MS(n) data. Quantification was done using their normalized and elemental specific response in the ICP-MS. The importance of these kinds of quantification strategies is stressed by the observation that the difference of the position of one oxygen atom in a structure can greatly affect its response in ESI-MS and UV detection. PMID- 21645662 TI - Characterization of drug-lysozyme conjugates by sheathless capillary electrophoresis-time-of-flight mass spectrometry. AB - Drug-protein conjugates have been widely used for the cell-specific targeting of drugs to cells that can bind and internalize the proteinaceous carrier. For renal drug targeting, lysozyme (LZM) can be used as an effective carrier that accumulates in proximal tubular cells. We used capillary electrophoresis-time-of flight mass spectrometry (CE-TOF-MS) for the characterization of different drug LZM conjugates. A recently developed prototype porous tip sprayer was employed for sheathless electrospray ionization (ESI) CE-MS interfacing. In order to prevent adsorption of LZM conjugates to the capillary wall, a positively charged polyethylenimine capillary coating was used in combination with a low-pH background electrolyte. Drug-LZM products had been prepared by first coupling BOC l-methionine hydroxysuccinimide ester (BOCmet) to lysine residues of LZM followed by conjugation with the kinase inhibitors LY364947, erlotinib, or Y27632 via a platinum(II)-based linker. CE-TOF-MS of each preparation showed narrow symmetrical peaks for the various reaction products demonstrating that drug-LZM conjugates remained stable during the CE analysis and subsequent ESI. Components observed in the drug-LZM products were assigned based on their relative migration times and on molecular mass as obtained by TOF-MS. The TOF-MS data obtained for the individual components revealed that the preparations contained LZM carrying one or two drug molecules, next to unmodified and BOCmet-modified LZM. Based on relative peak areas (assuming an equimolar response for each component) a quantitative conjugate profile could be derived for every preparation leading to drug loading values of 0.4-0.6 mol drug per mole protein. PMID- 21645663 TI - Determination of parabens in shampoo using high performance liquid chromatography with amperometric detection on a boron-doped diamond electrode. AB - Methylparaben (MePa), ethylparaben (EtPa) and propylparaben (PrPa) have been widely used, among others, as chemical preservatives in cosmetics, drugs and foods. As these compounds are linked with allergies, dermatitis and estrogenic properties, it is necessary to control the concentration of these substances in different matrices. The aim of this paper are: to evaluate the electrochemical behavior of parabens on the boron-doped diamond (BDD) electrode and the development of a chromatographic method, with electrochemical detection (HPLC ED), for determination of parabens in shampoo. A BDD (8000 ppm) electrode was adapted in a thin layer mode analytical cell consisting of a stainless steel and a platinum wire as reference and auxiliary electrodes, respectively. Chromatographic separations were obtained with a reversed phase C8 analytical column and a mobile phase of 0.025 molL(-1) disodium phosphate, pH 7.0, and acetonitrile (40:60, v/v), delivered at a flow rate of 1.0 mL min(-1). Sample preparation was performed by solid phase extraction using C18 cartridges and acetonitrile for elution. Benzylparaben was employed as internal standard. The HPLC-ED method developed, using the BDD electrode, was validated for the determination of parabens in shampoos and presented adequate linearity (>0.999), in the range of 0.0125-0.500% (w/w), detectability 0.01% (w/w), precision (RSD of 2.3-9.8%) and accuracy (93.1-104.4%) and could be applied for routine quality control of shampoos containing MePa, EtPa and PrPa. PMID- 21645664 TI - Review of the applications of different analytical techniques for coxibs research. AB - An extensive survey of the literature published in analytical and pharmaceutical chemistry journals has been conducted and analytical methods which were developed and used for the determination of some of the COX-2 inhibitors, a subclass of non steroidal anti-inflammatory drugs (NSAIDs) in bulk drugs, formulations, and biological fluids have been reviewed. This review covers the time period from 1999 to present, during which over 140 analytical procedures including chromatographic, spectrometric, electrophoretic and voltammetric techniques were reported. Presented applications concern analysis of coxibs from pharmaceutical formulations and biological samples. PMID- 21645665 TI - Integrating amperometric detection with electrophoresis microchip devices for biochemical assays: recent developments. AB - Recent advances in microfluidic systems, particularly in the Micro Total Analysis System (MUTAS) or Lab On a Chip (LOC), drive the current analysis tools and equipment towards miniaturization, rapid at-line testing and mobility. The state of-the-art microfluidic technology targets a wider range but smaller volumes of analytes, making the analytical procedure relatively easier and faster. This trend together with faster electronics and modern instrumentation systems will make real-time and in situ analysis a definite possibility. This review focuses on microchip capillary electrophoresis with amperometric detection (MCE-AD) for the detection of DNA and other electroactive analytes. The problems associated with the microchip design, in particular the choice of materials and the configuration of electrodes are discussed thoroughly and solutions are proposed. Significant developments in the related areas are also covered and reviewed critically. PMID- 21645666 TI - Biosensors elaborated on gold nanoparticles, a PM-IRRAS characterisation of the IgG binding efficiency. AB - This work is focused on studying the grafting of gold nanoparticles (Np) on a cystamine self-assembled monolayer on gold, in order to build sensitive immunosensors. The synthesis and deposition of gold nanoparticles, 13 and 55 nm sizes, were characterised by combining Polarisation Modulation Infrared Reflection-Absorption Spectroscopy (PM-IRRAS), X-ray Photoelectron Spectroscopy (XPS) Surface Enhanced Raman Scattering (SERS), and Atomic Force Microscopy (AFM) which all indicated the formation of a dispersed layer of nanoparticles. This observation is explained by the compromise between the high reactivity of amine terminated layers towards gold, and interparticle repulsions. Nps were then functionalised with antibody probes, and the recognition by an anti-rIgG was assayed both on planar and Np gold surfaces. The important result is that nanoparticles of 55 nm are preferable for the following reasons: they enable to build a denser and well dispersed layer and they increase both the number of receptors (IgGs) and their accessibility. Beside these geometric improvements, a net enhancement of the Raman signal was observed on the 55 nm nanoparticle layer, making this new platform promising for optical detection based biosensors. PMID- 21645667 TI - Bilayer lipid membrane biosensor with enhanced stability for amperometric determination of hydrogen peroxide. AB - In this paper, a polydopamine (PDA) film is electropolymerized on the surface of bilayer lipid membrane (BLM) which is immobilized with horseradish peroxidase (HRP). The coverage of the PDA film on HRP/BLM electrode is monitored by electrochemical impedance spectroscopy (EIS). The electrocatalytic reduction of H(2)O(2) at the PDA/HRP/BLM electrode is studied by means of cyclic voltammetry (CV). The biosensor has a fast response to H(2)O(2) of less than 5s and an excellent linear relationship is obtained in the concentration range from 2.5*10( 7) to 3.1*10(-3) molL(-1), with a detection limit of 1.0*10(-7) molL(-1) (S/N=3). The response current of BLM/HRP/PDA biosensor retains 84% of its original response after being stored in 0.1 molL(-1) pH 7.0 PBS at 4 degrees C for 3 weeks. The selectivity, repeatability, and storage stability of PDA/HRP/BLM biosensor are greatly enhanced by the coverage of polydopamine film on BLM. PMID- 21645668 TI - Measurement of methyl mercury (I) and mercury (II) in fish tissues and sediments by HPLC-ICPMS and HPLC-HGAAS. AB - A procedure for the extraction and determination of methyl mercury and mercury (II) in fish muscle tissues and sediment samples is presented. The procedure involves extraction with 5% (v/v) 2-mercaptoethanol, separation and determination of mercury species by HPLC-ICPMS using a Perkin-Elmer 3 MUm C8 (33 mm*3 mm) column and a mobile phase 3 containing 0.5% (v/v) 2-mercaptoethanol and 5% (v/v) CH(3)OH (pH 5.5) at a flow rate 1.5 ml min(-1) and a temperature of 25 degrees C. Calibration curves for methyl mercury (I) and mercury (II) standards were linear in the range of 0-100 MUgl(-1) (r(2)=0.9990 and r(2)=0.9995 respectively). The lowest measurable mercury was 0.4 MUgl(-1) which corresponds to 0.01 MUgg(-1) in fish tissues and sediments. Methyl mercury concentrations measured in biological certified reference materials, NRCC DORM - 2 Dogfish muscle (4.4+/-0.8 MUgg(-1)), NRCC Dolt - 3 Dogfish liver (1.55+/-0.09 MUgg(-1)), NIST RM 50 Albacore Tuna (0.89+/-0.08 MUgg(-1)) and IRMM IMEP-20 Tuna fish (3.6+/-0.6 MUgg(-1)) were in agreement with the certified value (4.47+/-0.32MUgg(-1), 1.59+/-0.12 MUgg(-1), 0.87+/-0.03 MUgg(-1), 4.24+/-0.27 MUgg(-1) respectively). For the sediment reference material ERM CC 580, a methyl mercury concentration of 0.070+/-0.002 MUgg(-1) was measured which corresponds to an extraction efficiency of 92+/-3% of certified values (0.076+/-0.04 MUgg(-1)) but within the range of published values (0.040-0.084 MUgg(-1); mean+/-s.d.: 0.073+/-0.05 MUgg(-1), n=40) for this material. The extraction procedure for the fish tissues was also compared against an enzymatic extraction using Protease type XIV that has been previously published and similar results were obtained. The use of HPLC-HGAAS with a Phenomenox 5 MUm Luna C18 (250 mm*4.6 mm) column and a mobile phase containing 0.06 moll(-1) ammonium acetate (Merck Pty Limited, Australia) in 5% (v/v) methanol and 0.1% (w/v) l-cysteine at 25 degrees C was evaluated as a complementary alternative to HPLC-ICPMS for the measurement of mercury species in fish tissues. The lowest measurable mercury concentration was 2 MUgl(-1) and this corresponds to 0.1 MUgg(-1) in fish tissues. Analysis of enzymatic extracts analysed by HPLC-HGAAS and HPLC-ICPMS gave equivalent results. PMID- 21645669 TI - Electrogenerated chemiluminescence of anatase TiO2 nanotubes film. AB - Highly ordered titanium dioxide (TiO(2)) nanotubes film was successfully synthesized via anodic oxidation of a Ti foil in an ammonium fluoride-based ethylene glycol solution. The electrogenerated chemiluminescence (ECL) behavior of the resulting TiO(2) nanotubes film was subsequently studied. Strong ECL emission was observed at -1.40V (vs. Ag/AgCl) and the ECL spectrum displayed three emission peaks which were bathochromatically shifted by ca. 140nm as compared to its corresponding photoluminescence (PL) emission peaks, indicating that the surface state plays an important role in the emission process. The ECL emission can also occur in a deareated solution attributing to the surface adsorbed O(2) molecules. The ECL emission intensity was quenched by dopamine and greatly enhanced in the presence of dissolved O(2) and H(2)O(2), making it possible to detect these analytes. The TiO(2) nanotubes film has been successfully applied to determine the dissolved O(2) content in river and pond water samples, the H(2)O(2) concentration in commercial disinfectant samples and the dopamine concentration in commercial dopamine injections with satisfactory results. The plausible ECL mechanisms of TiO(2) nanotubes film in aqueous solution are discussed. PMID- 21645670 TI - Gondola-shaped tetra-rhenium metallacycles modified evanescent wave infrared chemical sensors for selective determination of volatile organic compounds. AB - Water-stable and cavity-contained rhenium metallacycles were synthesized, and their ability to selectively interact with volatile organic compounds (VOCs) systematically studied using attenuated total reflection infrared (ATR-IR) spectroscopy. Integrating the unique properties of rhenium metallacycles into optical sensing technologies significantly improves selectivity in detecting aromatic compounds. To explore the interaction of rhenium metallacycles with VOCs, the surface of ATR sensing elements was modified with the synthesized rhenium metallacycles and used to detect VOCs. The results indicate that rhenium metallacycles have crown ether-like recognition sites, which can selectively interact with aromatic compounds, especially those bearing polar functional groups. The IR absorption bands of rhenium metallacycles shift significantly upon adsorption of aromatic VOCs, revealing a strong interaction between the tetra rhenium metallacycles and guest aromatic compounds. Optimizing the thickness of the metallacycles coated on the surface of the sensing element led to rapid response in detection. The dynamic range of response was generally up to 30 mg/L with detection limits ca. 30 MUg/L. Further studies of the effect of interferences indicate that recovery can be higher than 95% for most of the compounds tested. The results on the flow-cell device indicated that the performances were similar to a static detection system but the detection of VOCs can be largely simplified. PMID- 21645671 TI - Two-step protease digestion and glycopeptide capture approach for accurate glycosite identification and glycoprotein sequence coverage improvement. AB - A novel two-step protease digestion and glycopeptide capture approach has been developed. It is different from traditional tryptic digestion, glycopeptide enriching and identification approach in glycoproteomics. Here, proteins were first digested by Lys-C into relatively large peptides. Glycopeptides among them were selectively captured by hydrazide resin through oxidized glycans. After thorough washing steps, trypsin was used as a second protease to in situ release non-glycosylated part (named as LT-peptides) from glycopeptides. Subsequently, the remaining part of glycopeptides on resin was de-glycosylated by peptide-N glycosidase F, and collected as DG-peptides. Finally, both LT- and DG-peptides could be analyzed by mass spectrometer, achieving glycoprotein and glycosite identification. The approach was applied to cell lysate after positive validation by a model glycoprotein: 143 N-glycoproteins identified from DG- and LT-fraction both. In those glycoproteins, 189 DG-peptide-revealed N-glycosites got further confirmation by neighboring LT-peptides, which, in the meantime, made 109 glycoproteins get improved sequence coverage with increase even up to 350% (averagely 79.4%). Through controllable release, separate identification and combined interpretation of non-glycopeptides (newly introduced LT-peptides here) and traditional de-glycopeptides, the approach could not only achieve routine N glycosite identification, but also provide further proofs of N-glycosites and increase glycoprotein sequence coverage. PMID- 21645672 TI - Functionalization of graphene with electrodeposited Prussian blue towards amperometric sensing application. AB - Prussian blue (PB) was grown compactly on graphene matrix by electrochemical deposition. The as-prepared PB-graphene modified glassy carbon electrode (PB graphene/GCE) showed excellent electrocatalytic activity towards both the reduction of hydrogen peroxide and the oxidation of hydrazine, which could be attributed to the remarkable synergistic effect of graphene and PB. The PB graphene/GCE showed sensitive response to H(2)O(2) with a wide linear range of 10 1440 MUM at 0.0V, and to hydrazine with a wide linear range of 10-3000 MUM at 0.35 V. The detection limit was 3 MUM and 7 MUM, respectively, and both of them had rapid response within 5s to reach 95% steady state response. The wide linear range, good selectivity and long-time stability of the PB-graphene/GCE make it possible for the practical amperometric detection of hydrogen peroxide and hydrazine. PMID- 21645673 TI - Highly efficient rapid ethanol sensing based on Co-doped In2O3 nanowires. AB - Pristine and Co-doped In(2)O(3) nanowires were synthesized via electrospinning with subsequent calcination. Scanning electron microscopy, X-ray diffraction, and energy-dispersive X-ray spectroscopy were used to characterize nanowire morphology and structure. Ethanol sensing performance analyzed in the range of temperatures and concentrations showed that Co-doped In(2)O(3) nanowires exhibited significantly enhanced sensitivity and rate of performance with the response and recovery times of 2s and 3s, respectively. Combined with excellent selectivity and linearity, these properties make the fabricated nanowires a good candidate for practical ethanol sensing. Further performance improvements are possible with utilization of nanofiber continuity intrinsic of the used top-down nanowire nanomanufacturing process. PMID- 21645674 TI - Sweeping-micellar electrokinetic chromatography for the simultaneous analysis of tricyclic antidepressant and beta-blocker drugs in wastewater. AB - The simultaneous analysis of tricyclic antidepressant (amitriptyline, clomipramine, doxepin and nortriptyline) and beta-blocker (alprenolol, labetalol and propranolol) drugs in wastewater was developed via sweeping-micellar electrokinetic chromatography (MEKC) together with a simple liquid-liquid extraction step. For sweeping-MEKC, the amount of organic modifier in the separation electrolyte, the concentration of phosphoric acid in the sample matrix and the injection time of the sample were optimized. Sensitivity enhancements of up to 305-fold were achieved via sweeping. This allowed limits of detection (LOD) from 7 to 27 ng/mL. The relative standard deviations of migration time, corrected peak area and peak height were less than 3.2%, 7.8% and 4.5%, respectively. Liquid-liquid extraction using dichloromethane as solvent afforded up to 21-fold enrichment of the drugs from spiked wastewater. No interference of the sample matrix was observed and recoveries were obtained in the range of 77-113% for all analytes except labetalol at three spiking levels of 16, 80 and 160 ng/mL. Detection at the ng/mL level makes this simple, environmentally friendly and cost effective method competitive against recently reported methods using advanced liquid-phase separation techniques for monitoring similar drugs in wastewater. PMID- 21645675 TI - High sensitive and label-free colorimetric DNA detection based on nicking endonuclease-assisted activation of DNAzymes. AB - Horseradish peroxidase mimicking DNAzyme (HRP-DNAzyme) attracts growing interest as an amplifying label for biorecognition and biosensing events, especially for DNA detection. However, in the traditional designs, one target molecule can only generate one HRP-DNAzyme, which limits the signal enhancement and thus its sensitivity. In this article, we propose an amplified and label-free colorimetric DNA detection strategy based on nicking endonuclease (NEase)-assisted activation of HRP-DNAzymes (NEAA-DNAzymes). This new strategy relies on the hairpin-DNAzyme probe and NEase-assisted target recycling. In the hairpin-DNAzyme probe, the HRP DNAzyme sequence is protected in a "caged" inactive structure, whereas the loop region includes the target complementary sequence. Upon hybridization with target, the beacon is opened, resulting in the activation of the HRP-DNAzyme. Meanwhile, upon formation of the duplex, the NEase recognizes a specific nucleotide sequence and cleaves the hairpin-DNAzyme probe into two fragments. After nicking, the fragments of the hairpin-DNAzyme probe spontaneously dissociate from the target DNA. Amplification is accomplished by another hairpin DNAzyme probe hybridizing to the released intact target to continue the strand scission cycle, which results in activation of numerous DNAzymes. The activated HRP-DNAzymes generate colorimetric or chemiluminescence readout signals, thus providing the amplified detection of DNA. The detection limit of the colorimetric method is 10 pmol/L, which are three orders of magnitude lower than that without NEase. In addition, the detection limit of the chemiluminescence method is 0.2 pmol/L. Meanwhile, this strategy also exhibits high discrimination ability even against single-base mismatch. PMID- 21645676 TI - Molecularly imprinted stir bar sorptive extraction coupled with high performance liquid chromatography for trace analysis of sulfa drugs in complex samples. AB - A novel sulfamethazine molecularly imprinted polymer (MIP)-coated stir bar for sorptive extraction of eight sulfa drugs from biological samples was prepared. The MIP-coating was about 20 MUm thickness with the relative standard deviation (RSD) of 6.7% (n=10). It was characterized by scanning electron microscope, infrared spectrum, thermogravimetric analysis, and solvent-resistant investigation, respectively. The non-imprinted polymer (NIP)-coating was used for comparison. The adsorptive capacity and selectivity of MIP-coating were evaluated in detail. The MIP-coating showed higher adsorption capability and selectivity than the NIP-coating. The saturated adsorption amount of the MIP-coating was 4.6 times over that of the NIP-coating in toluene. Sulfamethazine could be detected after the MIP-coated stir bar sorptive extraction even at a low concentration of 0.2 MUg/L. The MIP-coating also exhibited selective adsorption ability to analogues of the template. A method for the determination of eight sulfa drugs in biological samples by MIP coated stir bar sorptive extraction coupled with high performance liquid chromatography (HPLC) was developed. The extraction conditions, including extraction solvent, extraction time, desorption solvent, desorption time and stirring speed, were optimized. The linear ranges were 1.0 100 MUg/L and 2.0-100 MUg/L for eight sulfonamides, respectively. The detection limits were within the range of 0.20-0.72 MUg/L. The method was successfully applied to simultaneous multi-residue analysis of eight sulfonamides in spiked pork, liver and chicken samples with the satisfactory recoveries. PMID- 21645677 TI - In situ chemo-synthesized multi-wall carbon nanotube-conductive polyaniline nanocomposites: characterization and application for a glucose amperometric biosensor. AB - A new glucose amperometric biosensor, based on electrodeposition of platinum nanoparticles onto the surface of multi-wall carbon nanotube (MWNT)-polyaniline (PANI) nanocomposites, and then immobilizing glucose oxidase (GOD) with covalent interaction and adsorption effect, was constructed in this paper. Firstly, the MWNT-PANI nanocomposites had been synthesized by in situ polymerization and were characterized through transmission electron microscopy (TEM), Fourier transform infrared (FT-IR) spectroscopy, and ultraviolet and visible (UV-vis) absorption spectra. The assembled process of the modified electrode was probed by scanning electron microscopy (SEM) and cyclic voltammetry (CV). Chronoamperometry was used to study the electrochemical performance of the resulting biosensor. The glucose biosensor exhibited a linear calibration curve over the range from 3.0 MUM to 8.2mM, with a detection limit of 1.0 MUM and a high sensitivity of 16.1 MUA mM( 1). The biosensor also showed a short response time (within 5s). Furthermore, the reproducibility, stability and interferences of the biosensor were also investigated. PMID- 21645678 TI - A polar-copolymerized method to prepare silica-based anion exchanger for ion chromatography. AB - A novel silica-based strong anion exchanger was developed for ion chromatography by copolymerizing methyltrichlorosilane and 3-chloropropyltrichlorosilane. The method allows the column capacity to be easy control simply by adjusting the ratio of silanes. The unwanted residual silanol groups onto the surface of silica gel could also be greatly reduced by this strategy. The effective column capacity of the column used was measured to be 50.8 MUequiv/column (2.03 MUequiv/cm). The exchanger was characterized by solid state CP/MAS (13)C NMR and elemental analysis and its separation performance was evaluated for the separation of common inorganic anions. The results showed that the column had good separation efficiency (e.g. the plate number of nitrite is 80,000/m) and the separation mechanism was observed to be dominantly governed by ion exchange mechanism. The utility of the column was demonstrated for the determination of nitrite and nitrate in saliva sample. PMID- 21645679 TI - A highly sensitive electrochemical immunosensor based on coral-shaped AuNPs with CHITs inorganic-organic hybrid film. AB - A highly sensitive electrochemical immunosensor based on combination of chitosan (CHIT) and coral-shaped AuNPs (C-AuNPs) to form an immobilization matrix has been developed using human IgG as a model analyte. The inorganic-organic hybrid film with abundant adsorbing sites and large surface area can reserve the biocompatibility of the biomaterials which greatly increase loading amounts of assembling, thus, significantly improves the performance of biosensing. The morphology is studied by scanning electron microscopy (SEM). Under the optimized experimental conditions, the immunosensor exhibits excellent performance (e.g., a detection limit of 5 pmol L(-1), a linear dynamic range of 3 orders of magnitude, high specificity). This possibly makes it an attractive platform for the direct immunoassay of human IgG or other biomolecules. PMID- 21645680 TI - Fast gradient high performance liquid chromatography method with UV detection for simultaneous determination of seven angiotensin converting enzyme inhibitors together with hydrochlorothiazide in pharmaceutical dosage forms and spiked human plasma and urine. AB - The development of a reversed phase liquid chromatographic method for the simultaneous determination of seven angiotensin converting enzyme (ACE) inhibitors; five drugs namely benazepril HCl (BZL), enalapril maleate (ENL), fosinopril sodium (FSP), lisinopril (LSP) and ramipril (RMP) and two metabolites captopril disulfide (CPD) and enalaprilat (ENT) together with hydrochlorothiazide (HCT) is described. The method can serve as a substitute for many published papers for the analysis of the targeted compounds with or without hydrochloothiazide in pharmaceutical formulations as well as in spiked human plasma and urine samples. The method utilizes a simple gradient procedure for the separation in a 11 min run time using acetonitrile aqueous ammonia buffer (pH 9) solution and an Extend RP-C18 (25 MUm particle size, 4.6 mm*250 mm, Agilent) HPLC column. The effluent was monitored on a UV detector at 215 nm. The effect of pH, solvent strength and analysis time on the peak shape and quantification were carefully studied in order to optimize the method. Adopting the proposed procedure, the analytes produce well-shaped peaks with good linear relationship over the investigated concentration ranges. The limits of detection (LOD) and limits of quantification (LOQ) from standard drug solutions lie in the range of 17-64 and 56-212 ng mL(-1), respectively. Correlation coefficient values (r) higher than 0.997 were obtained for all the studied drugs in spiked human plasma and urine samples. The intra-day and inter-day precision of the method was evaluated with relative standard deviation values being satisfactory for their purposed analysis. The method was validated with respect to specificity, recovery, accuracy, precision and linearity. PMID- 21645681 TI - Highly conducting gold nanoparticles-graphene nanohybrid films for ultrasensitive detection of carcinoembryonic antigen. AB - A new label-free amperometric immunosensor was developed for detection of carcinoembryonic antigen (CEA) based on chitosan-ferrocene (CS-Fc) and nano TiO(2) (CS-Fc+TiO(2)) complex film and gold nanoparticles-graphene (Au-Gra) nanohybrid. CS-Fc+TiO(2) composite membrane was first modified on a bare glass carbon electrode. Then Au-Gra nanohybrid was formed on the CS-Fc+TiO(2) membrane by self-assembly strategy. Next, further immobilization of anti-CEA was constructed according to the strong interaction between Au-Gra and the amido groups of anti-CEA. Since Au-Gra nanohybrid films provided a congenial microenvironment for the immobilization of biomolecules, the surface coverage of antibody protein could be enhanced and the sensitivity of the immunosensor has been improved. The good electronic conductive characteristic might be attributed to the synergistic effect of graphene nanosheets and Au NPs. The modified process was characterized by scanning electron microscope (SEM) and cyclic voltammetry (CV). Under optimized conditions, the resulting biosensor displayed good amperometric response to CEA with linear range from 0.01 to 80 ng/mL and a detection limit of 3.4 pg/mL (signal/noise=3). The results demonstrated that the immunosensor has advantages of high conduction, sensitivity, and long life time. This assay approach showed a great potential in clinical applications and detection of low level proteins. PMID- 21645682 TI - In situ spectral imaging of marker proteins in gastric cancer with near-infrared and visible quantum dots probes. AB - This study presents the investigation of bioconjugating ability of near-infrared (NIR) CdSeTe/ZnS quantum dots (QDs) (710 nm) and visible CdSe QDs (595 nm) in immunofluorescent staining for cancer biomarkers in gastric cancer tissues probed with the homemade Hadamard transform (HT) spectral imaging microscope and a commercial multispectral imaging system. The results show that imunostaining ability of NIR QDs probes is stronger than that of visible QDs when the two kinds of QDs are simultaneously used to probe the cancer biomarkers such as cytokeratin 20 (CK20) and proliferating cell nuclear antigen (PCNA) in gastric cancer tissues. Moreover, when the two QDs probes are used for immunostaining successively for the same target molecules, staining order has great influences on the final results due to their different conjugating ability to the marker proteins. The results imply that NIR QDs hold more promise for real-time imaging of tumor tissues due to its higher sensitivity and contrast. In addition, the results also demonstrate the potential of Hadamard transform spectral imaging as a useful tool in biomedical analysis and quantitative evaluation for tumor tissues. PMID- 21645683 TI - A novel ion-pairing chromatographic method for the simultaneous determination of both nicarbazin components in feed additives: chemometric tools for improving the optimization and validation. AB - The development, optimization and validation of an ion-pairing high performance liquid chromatography method for the simultaneous determination of both nicarbazin (NIC) components: 4,4'-dinitrocarbanilide (DNC) and 2-hydroxy-4,6 dimethylpyrimidine (HDP) in bulk materials and feed additives are described. An experimental design was used for the optimization of the chromatographic system. Four variables, including mobile phase composition and oven temperature, were analyzed through a central composite design exploring their contribution to analyte separation. Five responses: peak resolutions, HDP capacity factor, HDP tailing and analysis time, were modelled by using the response surface methodology and were optimized simultaneously by implementing the desirability function. The optimum conditions resulted in a mobile phase consisting of 10.0 mmol L(-1) of 1-heptanesulfonate, 20.0 mmol L(-1) of sodium acetate, pH=3.30 buffer and acetonitrile in a gradient system at a flow rate of 1.00 mL min(-1). Column was an INERSTIL ODS-3 (4.6 mm*150 mm, 5 MUm particle size) at 40.0 degrees C. Detection was performed at 300 nm by a diode array detector. The validation results of the method indicated a high selectivity and good precision characteristics, with RSD less than 1.0% for both components, both in intra and inter-assay precision studies. Linearity was proved for a range of 32.0-50.0 MUg mL(-1) of NIC in sample solution. The recovery, studied at three different fortification levels, varied from 98.0 to 101.4 for HDP and from 99.1 to 100.2 for DNC. The applicability of the method was demonstrated by determining DNC and HDP content in raw materials and commercial formulations used for coccidiosis prevention. Assays results on real samples showed that considerable differences in molecular ratio DNC:HDP exist among them. PMID- 21645684 TI - New approach to quantitative analysis of benzo[a]pyrene in food supplements by an immunochemical column test. AB - A quantitative immunochemical rapid test for sensitive determination of benzo[a]pyrene (BAP) as a model analyte was developed making use of a handheld reader for results evaluation. The covalent immobilization of antibodies to different Sepharose gels, i.e., CNBr-activated Sepharose 4B and CNBr-activated Sepharose 4 Fast Flow was compared with adsorption to a polyethylene support. The lowest limits of detection (LOD) were 4 ng L(-1) and 40 ng L(-1), respectively, using optimized assay conditions. The developed test was applied to food supplements (garlic, black radish and maca), including a pretreatment procedure. LOD of 9 ng kg(-1) and linear range of 13-80 ng kg(-1) were obtained. Results of BAP determination in naturally contaminated samples were confirmed by high performance liquid chromatography coupled to fluorescence detection and a good correlation was achieved. We suggest that the developed test format can be used to quantitative detection of the low molecular weight analytes, such as mycotoxins, pesticides, other pollutants in food and environmental samples. PMID- 21645685 TI - Development of a solvent-free method for the simultaneous identification/quantification of drugs of abuse and their metabolites in environmental water by LC-MS/MS. AB - This work details a rapid analytical method using direct sample injection for the simultaneous identification/quantification of 22 drugs of abuse, including some of their major metabolites, in environmental samples. This has been developed using a hybrid triple quadrupole-linear ion trap-mass spectrometer (QqLIT). With the increasing sensitivity of today's tandem mass spectrometers, direct injection analysis of water samples has become an attractive alternative to traditional analytical protocols, which often include a preliminary pre-concentration step. What's more, this kind of analysis is in accordance with many of the main objectives of so-called green analytical chemistry, or environmentally friendly practice. The analytical performance of the LC-MS/MS method was evaluated in three different water matrices (surface water, influent and effluent wastewater). Data acquisition was carried out in selected reaction monitoring (SRM) mode under time-scheduled conditions, monitoring two SRM transitions for simultaneous identification/quantification of all target compounds in the samples. Additionally, an experiment was performed using the information-dependent acquisition (IDA) scan to carry out the identification of those analytes for which the second transition was present at a low intensity. Finally, the two methodologies developed were applied to real samples for evaluation. PMID- 21645686 TI - Storage method, drying processes and extraction procedures strongly affect the phenolic fraction of rosemary leaves: an HPLC/DAD/MS study. AB - The Rosmarinus officinalis L. is widely known for its numerous applications in the food field but also for the increasing interest in its pharmaceutical properties. Two groups of compounds are mainly responsible for the biological activities of the plant: the volatile fraction and the phenolic constituents. The latter group is mainly constituted by rosmarinic acid, by a flavonoidic fraction and by some diterpenoid compounds structurally derived from the carnosic acid. The aim of our work was to optimize the extractive and analytical procedure for the determination of all the phenolic constituents. Moreover the chemical stability of the main phenols, depending on the storage condition, the different drying procedures and the extraction solvent, have been evaluated. This method allowed to detect up to 29 different constituents at the same time in a relatively short time. The described procedure has the advantage to being able to detect and quantify several classes of compounds, among them numerous minor flavonoids, thus contributing to improving knowledge of the plant. The findings from this study have demonstrated that storing the raw fresh material in the freezer is not appropriate for rosemary, mainly due to the rapid disappearing of the rosmarinic acid during the freezing/thawing process. Regarding the flavonoidic fraction, consistent decrements, were highlighted in the dried samples at room temperature if compared with the fresh leaf. Rosmarinic acid, appeared very sensitive also to mild drying processes. The total diterpenoidic content undergoes to little changes when the leaves are freeze dried or frozen and limited losses are observed working on dried leaves at room temperature. Nevertheless it can be taken in account that this fraction is very sensitive to the water presence during the extraction that favors the conversion of carnosic acid toward it oxidized form carnosol. From our findings, it appear evident that when evaluating the phenolic content in rosemary leaves, several factors, mainly the type of storage, the drying process and the extraction methods, should be carefully taken into account because they can induce partial losses of the antioxidant components. PMID- 21645687 TI - Olive oil quantification of edible vegetable oil blends using triacylglycerols chromatographic fingerprints and chemometric tools. AB - The present work studies the effectiveness of the use of triacylglycerols (TAGs) for the quantification of olive oil in blends with vegetable oils. The determinations were obtained using high-performance liquid chromatography (HPLC) coupled to a Charged Aerosol Detector (CAD), in combination with Partial Least Squares (PLS) regression and using interval PLS (iPLS) for variable selection. Results revealed that PLS models can predict olive oil concentrations with reasonable errors. Variable selection through iPLS did not improve predictions significantly, but revealed the chemical information important in the chromatogram to quantify olive oil in vegetable oil blends. PMID- 21645688 TI - Single solid phase extraction method for the simultaneous analysis of polar and non-polar pesticides in urine samples by gas chromatography and ultra high pressure liquid chromatography coupled to tandem mass spectrometry. AB - A new multiresidue method has been developed and validated for the simultaneous extraction of more than two hundred pesticides, including non-polar and polar pesticides (carbamates, organochlorine, organophosphorous, pyrethroids, herbicides and insecticides) in urine at trace levels by gas and ultra high pressure liquid chromatography coupled to ion trap and triple quadrupole mass spectrometry, respectively (GC-IT-MS/MS, UHPLC-QqQ-MS/MS). Non-polar and polar pesticides were simultaneously extracted from urine samples by a simple and fast solid phase extraction (SPE) procedure using C(18) cartridges as sorbent, and dichloromethane as elution solvent. Recovery was in the range of 60-120%. Precision values expressed as relative standard deviation (RSD) were lower than 25%. Identification and confirmation of the compounds were performed by the use of retention time windows, comparison of spectra (GC-amenable compounds) or the estimation of the ion ratio (LC-amenable compounds). For GC-amenable pesticides, limits of detection (LODs) ranged from 0.001 to 0.436 MUg L(-1) and limits of quantification (LOQs) from 0.003 to 1.452 MUg L(-1). For LC-amenable pesticides, LODs ranged from 0.003 to 1.048 MUg L(-1) and LOQs ranged from 0.011 to 3.494 MUg L(-1). Finally, the optimized method was applied to the analysis of fourteen real samples of infants from agricultural population. Some pesticides such as methoxyfenozide, tebufenozide, piperonyl butoxide and propoxur were found at concentrations ranged from 1.61 to 24.4 MUg L(-1), whereas methiocarb sulfoxide was detected at trace levels in two samples. PMID- 21645689 TI - Production of monoclonal antibodies against hop-derived (Humulus lupulus L.) prenylflavonoids and the development of immunoassays. AB - Monoclonal antibodies against the hop-derived prenylated chalcone xanthohumol (X) and the prenylated flavonoids isoxanthohumol (IX) and 8-prenylnaringenin (8-PN) were developed. Carboxylic acid haptens of X, IX and 8-PN were synthesized by linking a spacer to their C4'-OH group followed by subsequent coupling to bovine serum albumin (BSA) to form conjugates that were employed as immunogens in BALB/c mice to raise antibodies. The monoclonal antibodies that were secreted from the established hybridoma cell lines proved, in cross-reactivity studies, to possess highly specific binding capacities in an optimized competitive indirect ELISA. The immunoassays make use of immunogen-coated microtiterplates and a peroxidase labeled anti-mouse IgG(1) secondary antibody with ABTS as a chromogenic substrate. For X the IC(50) value derived from the standard curve was 62.91 ng mL(-1), and for both IX and 8-PN 37.15 ng mL(-1). The assay was validated for the quantitative analysis of X, IX and 8-PN in urine and serum. A simple sample pretreatment procedure using a diethyl ether extraction was optimized and the recoveries and matrix effects were assessed. The validity of the established assay was tested and mean inter- and intra-assay variations in urine were 2.32% and 1.91%, respectively for X, 6.24% and 2.39%, respectively for IX and 7.18% and 0.74%, respectively for 8-PN. In serum, the mean inter- and intra-assay variations were 8.90% and 1.37%, respectively for X, 6.13% and 1.57%, respectively for IX and 6.13% and 2.43%, respectively for 8-PN. Furthermore, the method demonstrated excellent accuracy and significant correlation with measurements by an established and validated HPLC-MS method. PMID- 21645690 TI - Evaluation of matrix solid-phase dispersion (MSPD) extraction for multi-mycotoxin determination in different flours using LC-MS/MS. AB - An existing matrix solid-phase dispersion (MSPD) method for aflatoxins (AFs) and ochratoxin A (OTA) extraction was extended by further 14 mycotoxins. After it careful optimization, this method was applied to determine the occurrence of these mycotoxins on commercial flour samples (with different cereals composition) collected from local markets. In a total of 49 samples investigated, 9 mycotoxins were identified. Nivalenol (NIV) and Beauvericin (BEA) were the mycotoxins found most frequently. The samples that presented major contamination were wheat flours and bakery preparations. Despite of the great number of positives finding, only one wheat flour sample exceeded the maximum limits (ML) for OTA established by the European Union (EU). However, it would be interesting to calculate the total ingest of these mycotoxins along the years. PMID- 21645691 TI - Acetylcholinesterase biosensor based on single-walled carbon nanotubes--Co phtalocyanine for organophosphorus pesticides detection. AB - A simple and reliable technique has been developed for the construction of an amperometric acetylcholinesterase biosensor based on screen-printed carbon electrodes. For the first time, one-step modification using single-walled carbon nanotubes and Co phtalocyanine has been proposed to decrease the working potential and to increase the signal of thiocholine oxidation. The biosensor developed made it possible to detect 5-50 ppb of paraoxon and 2-50 ppb of malaoxon with detection limits of 3 and 2 ppb, respectively (incubation 15 min). The biosensor showed high reproducibility when measurements of the substrate and inhibitor were performed (R.S.D. about 1% and 2.5%, respectively). The reliability of the inhibition measurements was confirmed by testing spiked samples of sparkling and tape waters. PMID- 21645692 TI - Gas chromatography - optical fiber detector for assessment of fatty acids in urban soils. AB - Fatty acids have been used as biomarkers of the microbial community composition of soils and they are usually separated and quantified by gas-chromatography coupled to a flame ionization detector (GC-FID). The aim of this study was to develop, validate and apply a methodology based on gas chromatography coupled to optical fiber detection (GC-OF) for screening five fatty acids used as indicators of fungal and bacterial communities in urban soils. The performance of the GC-OF methodology (optical fiber detector at 1,550 nm) was evaluated by comparison with the GC-FID methodology and it was found that they were comparable in terms of linear range, detection limit and analytical errors. Besides these similar analytical characteristics, the GC-OF is much cheaper than the GC-FID methodology. Different concentrations were determined for each fatty acid indicator which in turn varied significantly between the soil samples analyzed from Lisbon ornamental gardens. Additionally, the GC-OF showed a great potential as alternative for determination of eleven or more fatty acids in urban soils. PMID- 21645693 TI - Selective chemiluminescence method for monitoring of vitamin K homologues in rheumatoid arthritis patients. AB - Vitamin K is a fat-soluble vitamin involved in blood coagulation and bone metabolism. The detection and monitoring of vitamin K homologues in rheumatoid arthritis (RA) patients is a challenging problem due to the smaller concentrations of vitamin K and the presence of several interfering medications. Therefore, this study aimed to develop a new highly sensitive and selective chemiluminescence (CL) method designated to quantify vitamin K homologues in plasma of RA patients including phylloquinone (PK, vitamin K(1)), menaquinone-4 (MK-4, vitamin K(2)) and menaquinone-7 (MK-7, vitamin K(2)). The method was based on the unique photochemical properties of vitamin K homologues that were exploited for selective luminol CL reaction. The correlation coefficients of 0.998 or more were obtained in the concentration ranges of 0.1-100 ng mL(-1) vitamin K homologues. The detection limits were 0.03-0.1 ng mL(-1) in human plasma for vitamin K homologues. The developed HPLC-CL system was successfully applied for selective determination of vitamin K homologues in plasma of RA patients. The developed method may provide a useful tool for monitoring vitamin K homologues in different clinical studies such as RA, osteoporosis and hepatocellular carcinoma in which vitamin K is intervented. PMID- 21645694 TI - Chemometrics optimization of carbohydrate separations in six food matrices by micellar electrokinetic chromatography with anionic surfactant. AB - Multivariate statistical design modeling and the Derringer-Suich desirability function analysis were applied to micellar electrokinetic chromatography (MEKC) results with anionic surfactant to separate carbohydrates (CHOs) in different food matrices. This strategy has been studied with success to analyze compounds of difficult separation, but has not been explored for carbohydrates. Six procedures for the analysis of different sets of CHOs present in six food matrices were developed. The effects of pH, electrolyte and surfactant concentrations on the separation of the compounds were investigated using a central composite design requiring 17 experiments. The simultaneous optimization of the responses for separation of six sets of CHOs was performed employing empirical models for prediction of optimal resolution conditions in six matrices, condensed milk, orange juices, rice bran, red wine, roasted and ground coffee and breakfast cereal samples. The results indicate good separation for the samples, with appropriate detectability and selectivity, short analysis time, low reagent cost and little waste generation, demonstrating that the proposed technique is a viable alternative for carbohydrate analysis in foods. PMID- 21645695 TI - On-line solid phase extraction of Ni and Pb using carbon nanotubes and modified carbon nanotubes coupled to ETAAS. AB - A study about the capabilities of three kinds of nanomaterials namely, carbon nanotubes (CNT), oxidized carbon nanotubes (ox-CNT) and l-alanine immobilized on carbon nanotubes (ala-CNT) to serve as sorbents for preconcentrating Ni and Pb using an on-line system coupled to electrothermal atomic absorption spectroscopy (ETAAS) technique, was accomplished. The solid phase extraction was performed in a conical minicolumn used as sorbent holder. After loading a fixed volume of the analytes, they were eluted with a discrete volume (50 MUL) of nitric acid, placed directly into the platform of a L'Vov tube. After that, each analyte was determined individually. Ni and Pb retention was strongly influenced by pH but exhibiting different behaviors. The study demonstrated that the sorbent based on ox-CNTs was the one that exhibited the highest capacity and linearity for both analytes when compared with CNT or ala-CNT. The analytical performance was evaluated for the three sorbents to establish the best conditions regarding sensitivity, reproducibility and accuracy. The precision expressed as relative standard deviations (n=6) were 3.9 and 0.5% for Ni(2+) and Pb(2+), respectively The limit of detection (LOD), calculated as the concentration required to yield a net peak equal to three times the standard deviation of the background signal (3sigma) was 30 and 10 ng L(-1) for Ni(2+) and Pb(2+) respectively. Alternatively, the limit of quantification (10sigma) was calculated and resulted to be 0.79 and 0.07 MUg L(-1) for Ni(2+) and Pb(2+) respectively. After optimization, the method that involved the use of ox-CNT associated to an on-line preconcentration was tested in samples of relevant environmental importance. Accuracy was evaluated analyzing a certified reference material namely, Municipal Sludge (QC MUNICIPAL SLUDGE A) and a reference sample of Lake Sediment (TRAP-LRM from IJS). PMID- 21645696 TI - Determination of elaidic and vaccenic acids in foods using GC*GC-FID and GC*GC TOFMS. AB - Trans fatty acids (TFAs) are present in meat and dairy products as m ruminant animals and in vegetable fats due to partial hydrogenation. This study aimed to discriminate between natural (N-TFA) and hydrogenated trans fatty (H-TFA) acids by GC*GC-flame ionization detection (GC*GC-FID) and comprehensive GC*GC-time-of flight mass spectrometry (GC*GC-TOFMS). The separation of two kinds of trans fats, vaccenic acid (18:1 trans-11) and elaidic acid (18:1 trans-9), was performed using GC*GC-FID and GC*GC-TOFMS. A 100 m*0.25 mm I.D.*0.2 MUm (film thickness) SP-2560 (bis-cyanopropyl polysiloxane) fused capillary column (first separation dimension, 1D) was coupled to a 1.5 m*0.18 mm I.D.*0.18 MUm (film thickness) RTX-5 (5% diphenyl/95% dimethyl polysiloxane) fused capillary column (second separation dimension, 2D). The RSD of the intra-day repeatability by both GC*GC-FID and GC*GC-TOFMS for elaidic and vaccenic acids was <=9.56% and <=9.97%, and the RSD of the inter-day repeatability was <=8.49 and <=9.06%, respectively. It was found that the V/E value (vaccenic acid to elaidic acid ratio) could be used to distinguish H-TFA from N-TFA and to evaluate the quality of the fatty foods. PMID- 21645697 TI - Zone trapping/merging zones in flow analysis: a novel approach for rapid assays involving relatively slow chemical reactions. AB - A novel strategy for accomplishing zone trapping in flow analysis is proposed. The sample and the reagent solutions are simultaneously inserted into convergent carrier streams and the established zones merge together before reaching the detector, where the most concentrated portion of the entire sample zone is trapped. The main characteristics, potentialities and limitations of the strategy were critically evaluated in relation to an analogous flow system with zone stopping. When applied to the spectrophotometric determination of nitrite in river waters, the main figures of merit were maintained, exception made for the sampling frequency which was calculated as 189 h(-1), about 32% higher relatively to the analogous system with zone stopping. The sample inserted volume can be increased up to 1.0 mL without affecting sampling frequency and no problems with pump heating or malfunctions were noted after 8-h operation of the system. In contrast to zone stopping, only a small portion of the sample zone is halted with zone trapping, leading to these beneficial effects. PMID- 21645698 TI - Multivariate curve resolution modeling of liquid chromatography-mass spectrometry data in a comparative study of the different endogenous metabolites behavior in two tomato cultivars treated with carbofuran pesticide. AB - A metabonomic study based on the application of multivariate curve resolution and alternating least squares (MCR-ALS) to three-way data sets obtained by liquid chromatography coupled to mass spectrometry detection (LC-MS) was carried out for Rambo and Raf tomato cultivars treated with carbofuran pesticide. Samples were picked up during a 21 days period after treatment and analyzed by LC-MS in scan mode, along with the corresponding blank samples. Then, MCR-ALS was applied to the three-way data sets using column wise augmented matrices, and the evolutionary profiles as a function of the time after treatment were estimated for the metabolites present in both cultivars, as well as their corresponding pure spectra estimations. A comparative study using those estimations showed that some of these metabolites followed different behavior for the different cultivars after treatment. Since all treated and untreated Rambo and Raf samples were picked up according to the same sampling protocol and in a similar state of maturation, any difference in the behavior between profiles can be interpreted as an effect due to the presence of pesticide and to the kind of cultivar. Based on this hypothesis, several PLS-DA approaches were tested to check if it would be possible to classify samples by using the metabolites MCR estimations. Results showed that PLS-DA models for classification of treated or non-treated (blank) samples were the best ones obtained (98.44% of correct classifications for the validation set), which supports the stress effects related to carbofuran treatment. In addition, excellent discrimination among the four groups could be attained (89.06% of correct classifications for the validation set). PMID- 21645699 TI - Rapid detection of Escherichia coli contamination in packaged fresh spinach using hyperspectral imaging. AB - A rapid method based on hyperspectral imaging for detection of Escherichia coli contamination in fresh vegetable was developed. E. coli K12 was inoculated into spinach with different initial concentrations. Samples were analyzed using a colony count and a hyperspectroscopic technique. A hyperspectral camera of 400 1000 nm, with a spectral resolution of 5 nm was employed to acquire hyperspectral images of packaged spinach. Reflectance spectra were obtained from various positions on the sample surface and pretreated using Sawitzky-Golay. Chemometrics including principal component analysis (PCA) and artificial neural network (ANN) were then used to analyze the pre-processed data. The PCA was implemented to remove redundant information of the hyperspectral data. The ANN was trained using Bayesian regularization and was capable of correlating hyperspectral data with number of E. coli. Once trained, the ANN was also used to construct a prediction map of all pixel spectra of an image to display the number of E. coli in the sample. The prediction map allowed a rapid and easy interpretation of the hyperspectral data. The results suggested that incorporation of hyperspectral imaging with chemometrics provided a rapid and innovative approach for the detection of E. coli contamination in packaged fresh spinach. PMID- 21645700 TI - Molecularly imprinted polymers by reversible addition-fragmentation chain transfer precipitation polymerization for preconcentration of atrazine in food matrices. AB - Controlled/living free radical polymerization (CLRP) has been accepted as an effective technique in preparation of polymers because of its inherent advantages over traditional free radical polymerization. In this work, reversible addition fragmentation chain transfer (RAFT) polymerization, the ideal candidate for CLRP, was applied to prepare atrazine molecularly imprinted polymers (MIPs) by precipitation polymerization. The resultant RAFT-MIPs demonstrated uniform spherical shape with rough surface containing significant amounts of micropores, leading to an improvement in imprinting efficiency compared with that of the MIPs prepared by traditional precipitation polymerization (TR-MIPs). The maximum binding capacities of the RAFT-MIPs and TR-MIPs were 2.89 mg g(-1) and 1.53 mg g( 1), respectively. The recoveries ranging from 81.5% to 100.9% were achieved by one-step extraction by using RAFT-MIPs for preconcentration and selective separation of atrazine in spiked lettuce and corn samples. These results provided the possibility for the separation and enrichment of atrazine from complicated matrices by RAFT-MIPs. PMID- 21645701 TI - Hg(II) immobilized MWCNT graphite electrode for the anodic stripping voltammetric determination of lead and cadmium. AB - The preparation of Hg(II)-modified multi walled carbon nanotube (MWCNT) by reaction of oxidized MWCNT with aqueous HgCl(2) was carried out. The Hg(II) modified multi walled carbon nanotube (Hg(II)/MWCNT) dispersed in Nafion solution was used to coat the polished graphite electrode surface. The Hg(II)/MWCNT modified graphite electrode was held at a cathodic potential (-1.0 V) to reduce the coordinated Hg(II) to Hg forming nanodroplets of Hg. The modified electrode was characterized by FESEM/EDAX which provided useful insights on the morphology of the electrode. The SEM images showed droplets of Hg in the size of around 260 nm uniformly distributed on the MWCNT. Differential pulse anodic stripping voltammetry (DPASV) and electrochemical impedance spectroscopy were used to study the Hg(II) binding with MWCNT. Differential pulse anodic stripping voltammetry of ppb levels of cadmium and lead using the modified electrode yielded well-defined peaks with low background current under a short deposition time. Detection limit of 0.94 and 1.8 ng L(-1) were obtained following a 3 min deposition for Pb(II) and Cd(II), respectively. Various experimental parameters were characterized and optimized. High reproducibility was observed from the RSD values for 20 repetitive measurements of Pb(II) and Cd(II) (1.7 and 1.9%, respectively). The determination of Pb(II) and Cd(II) in tap water and Pb(II) in human hair samples was carried out. The above method of fabrication of Hg(II)/MWCNT modified graphite electrode clearly suggests a safe route for preparing Hg immobilized electrode for stripping analysis. PMID- 21645702 TI - UV-ablation nanochannels in micro/nanofluidics devices for biochemical analysis. AB - This paper presents a simple and cost-effective UV-ablation technique for fabrication of size-tunable nanofluidics devices via photochemical decomposition reaction. UV-irradiation through a PET photomask results in continuous decomposition of poly(carbonate) (PC), forming nanochannel and carboxyl groups on the surface of the etched PC. This photochemical decomposition process occurs at molecular scale, therefore, the depth of nanochannels can be controlled at nanometer level. The etching rate is estimated to be ca. 0.015 nms(-1). To demonstrate the potential application of the present UV-ablation technique, a nanochannel was fabricated and integrated with microchannels to form a micro/nanofluidics chip for protein concentration. Using this device, about 10(3) 10(5) fold protein concentration can be achieved within 10 min. The present approach offers a simple and practical solution to fabricate nanofluidics devices at low-cost, and the resulting device could provide ideal platforms for MUTAS towards various applications in biology and chemistry. PMID- 21645703 TI - Integration of carboxyl modified magnetic particles and aqueous two-phase extraction for selective separation of proteins. AB - Both of the magnetic particle adsorption and aqueous two-phase extraction (ATPE) were simple, fast and low-cost method for protein separation. Selective proteins adsorption by carboxyl modified magnetic particles was investigated according to protein isoelectric point, solution pH and ionic strength. Aqueous two-phase system of PEG/sulphate exhibited selective separation and extraction for proteins before and after magnetic adsorption. The two combination ways, magnetic adsorption followed by ATPE and ATPE followed by magnetic adsorption, for the separation of proteins mixture of lysozyme, bovine serum albumin, trypsin, cytochrome C and myloglobin were discussed and compared. The way of magnetic adsorption followed by ATPE was also applied to human serum separation. PMID- 21645704 TI - A disposable electrochemical sensor for the determination of indole-3-acetic acid based on poly(safranine T)-reduced graphene oxide nanocomposite. AB - A disposable electrochemical sensor for the determination of indole-3-acetic acid (IAA) based on nanocomposites of reduced graphene oxide (rGO) and poly(safranine T) (PST) was reported. The sensor was prepared by coating a rGO film on a pre anodized graphite electrode (AGE) through dipping-drying and electrodepositing a uniform PST layer on the rGO film. Scanning electron microscopic (SEM) and infrared spectroscopic (IR) characterizations indicated that PST-rGO formed a rough and crumpled composite film on AGE, which exhibited high sensitive response for the oxidation of IAA with 147-fold enhancement of the current signal compared with bare AGE. The voltammetric current has a good linear relationship with IAA concentration in the range 1.0*10(-7)-7.0*10(-6)M, with a low detection limit of 5.0*10(-8)M. This sensor has been applied to the determination of IAA in the extract samples of several plant leaves and the recoveries varied in the range of 97.71-103.43%. PMID- 21645705 TI - Investigation on the preparation and chromatographic behavior of a new para-tert butylcalix[4]arene-1,2-crown-4 stationary phase for high performance liquid chromatography. AB - In the present work, a new para-tert-butylcalix[4]arene-1,2-crown-4 bonded silica stationary phase (CBS4-4) was synthesized, structurally characterized, and employed to separate polycyclic aromatic hydrocarbons (PAHs), phenols, aromatic amines, benzoic acid and its derivatives. The chromatographic behaviors of the prepared stationary phase were investigated and compared with ODS. The effects of methanol concentrations on the retention index show that CBS4-4 exhibits high selectivity for the above analytes. The separation mechanisms based on the different interactions between calixarene and the analytes were discussed. With the assistance of quantum chemistry calculation, the interaction Gibbs free energy change DeltaG(solv) (in the mobile phase) of p, m and o-phenylenediamine positional isomers and para-tert-butylcalix[4]arene-1,2-crown-4 were obtained. The DeltaG(solv) values were consistent with the retention behavior of p, m and o phenylenediamine on the CBS4-4. According to the chromatographic data, it can be concluded that the selectivity of CBS4-4 for analytes is mainly ascribed to hydrophobic interaction, accompanied by other effects such as hydrogen bonding interaction, pi-pi and inclusion interaction. The CBS4-4 column has been successfully employed for the analysis of benzoic acid in Sprite drink. PMID- 21645706 TI - The maintenance of the second-order advantage: second-order calibration of excitation-emission matrix fluorescence for quantitative analysis of herbicide napropamide in various environmental samples. AB - A rapid non-separative spectrofluorometric method based on the second-order calibration of excitation-emission matrix (EEM) fluorescence was proposed for the determination of napropamide (NAP) in soil, river sediment, and wastewater as well as river water samples. With 0.10 mol L(-1) sodium citrate-hydrochloric acid (HCl) buffer solution of pH 2.2, the system of NAP has a large increase in fluorescence intensity. To handle the intrinsic fluorescence interferences of environmental samples, the alternating penalty trilinear decomposition (APTLD) algorithm as an efficient second-order calibration method was employed. Satisfactory results have been achieved for NAP in complex environmental samples. The limit of detection obtained for NAP in soil, river sediment, wastewater and river water samples were 0.80, 0.24, 0.12, 0.071 ng mL(-1), respectively. Furthermore, in order to fully investigate the performance of second-order calibration method, we test the second-order calibration method using different calibration approaches including the single matrix model, the intra-day various matrices model and the global model based on the APTLD algorithm with nature environmental datasets. The results showed the second-order calibration methods also enable one or more analyte(s) of interest to be determined simultaneously in the samples with various types of matrices. The maintenance of second-order advantage has been demonstrated in simultaneous determinations of the analyte of interests in the environmental samples of various matrices. PMID- 21645707 TI - Fast haptoglobin phenotyping based on microchip electrophoresis. AB - A new and fast method for haptoglobin phenotyping was developed based on microchip electrophoresis with laser induced fluorescence detection. Haptoglobin phenotypes 1-1 and 2-2 were labeled with fluorescein isothiocyanate. The analyses were performed on glass microchip which was simply treated with sodium dodecyl sulfate. After the optimization of the separation conditions, Hp 1-1 and Hp 2-2 could be differentiated in 150s and the detection limits for Hp 1-1 and Hp 2-2 were 0.39 and 0.62 MUg/mL, respectively. Finally, the method was applied to human serum samples from healthy people and liver cancer patients. A decrease in Hp concentration for liver cancer patients was confirmed. Featuring high efficiency, speed, simplicity, the method reveals great potentials for the diagnosis of diseases and proteome research. PMID- 21645708 TI - Mechanism study on inorganic oxidants induced inhibition of Ru(bpy)32+ electrochemiluminescence and its application for sensitive determination of some inorganic oxidants. AB - Inhibited Ru(bpy)(3)(2+) electrochemiluminescence by inorganic oxidants is investigated. Results showed that a number of inorganic oxidants can quench the ECL of Ru(bpy)(3)(2+)/tri-n-propylamine (TPrA) system, and the logarithm of the decrease in ECL intensity (DeltaI) was proportional to the logarithm of analyte concentrations. Based on which, a sensitive approach for detection of these inorganic oxidants was established, e.g. the log-log plots of DeltaI versus the concentration of MnO(4)(-), Cr(2)O(7)(2-) and Fe(CN)(6)(3-) are linear in the range of 1*10(-7) to 3*10(-4)M for MnO(4)(-) and Cr(2)O(7)(2-), and 1*10(-7) to 1*10(-4)M for Fe(CN)(6)(3-), with the limit of detection (LOD) of 8.0*10(-8)M, 2*10(-8)M, and 1*10(-8)M, respectively. A series of experiments such as a comparison of the inhibitory effect of different compounds on Ru(bpy)(3)(2+)/TPrA ECL, ECL emission spectra, UV-Vis absorption spectra etc. were investigated in order to discover how these inorganic analytes quench the ECL of Ru(bpy)(3)(2+)/TPrA system. A mechanism based on consumption of TPrA intermediate (TPrA(.)) by inorganic oxidants was proposed. PMID- 21645709 TI - Quantitative detection of trace perfluorinated compounds in environmental water samples by Matrix-assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry with 1,8-bis(tetramethylguanidino)-naphthalene as matrix. AB - Determination of perfluorinated compounds (PFCs) is very important because of their potential hazards to the environment and human health. In present work, 1,8 bis (tetramethylguanidino)-naphthalene (TMGN), a superbasic proton sponge, was firstly employed as the matrix for quantitative detection of acidic PFCs in environmental water samples by Matrix-assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry (MALDI-TOF-MS). Several acidic PFCs, such as perfluorooctanesulfonate (PFOS) and perfluorooctanoic acid (PFOA), were selected as model analytes for demonstrating the feasibility of the detection method. The results showed that deprotonated ions of these PFCs were detected without any other matrix ions interference. The achieved sensitivity with TMGN for PFOS detection was ten-fold higher than that with 1,8-bis (dimethyl-amino)-naphthalene (DMAN) which was used for the detection of fatty acid by MALDI-TOF-MS. The high sensitivity of this method made it feasible to monitor and quantify acidic PFCs in complicated environmental water samples. Furthermore, a novel combined strategy of solid phase extraction (SPE) followed by MALDI-TOF-MS detection was developed for quantifying PFCs in environmental water samples. The calibration curves with a wide linear dynamic range (0.1-10 ng L(-1) for PFOS, PFHxS, and PFBS, and 0.5-50 ng L(-1) for PFOA, PFNA, and PFDA.) were obtained. The limit of detection (LOD) for PFOS of this method was 0.015 ng L(-1) (a signal-to-noise ratio of 3), which was lower than the LOD (0.036 ng L(-1)) obtained by high pressure liquid chromatography/tandem mass spectrometry (LC-MS/MS) method. Moreover, the strategy was used to detect the selected PFCs in water samples collected from Xiaoqinghe river and Gaobeidian wastewater. The achieved concentrations of PFCs were closed to those obtained by LC-MS/MS method. It is indicated that the proposed MALDI-TOF-MS method with TMGN as the matrix is much reliable and can be used as an alternative method to detect trace PFCs in environmental water samples. PMID- 21645710 TI - Optimization of flavanones extraction by modulating differential solvent densities and centrifuge temperatures. AB - Understanding the factors influencing flavonone extraction is critical for the knowledge in sample preparation. The present study was focused on the extraction parameters such as solvent, heat, centrifugal speed, centrifuge temperature, sample to solvent ratio, extraction cycles, sonication time, microwave time and their interactions on sample preparation. Flavanones were analyzed in a high performance liquid chromatography (HPLC) and later identified by liquid chromatography and mass spectrometry (LC-MS). The five flavanones were eluted by a binary mobile phase with 0.03% phosphoric acid and acetonitrile in 20 min and detected at 280 nm, and later identified by mass spectral analysis. Dimethylsulfoxide (DMSO) and dimethyl formamide (DMF) had optimum extraction levels of narirutin, naringin, neohesperidin, didymin and poncirin compared to methanol (MeOH), ethanol (EtOH) and acetonitrile (ACN). Centrifuge temperature had a significant effect on flavanone distribution in the extracts. The DMSO and DMF extracts had homogeneous distribution of flavanones compared to MeOH, EtOH and ACN after centrifugation. Furthermore, ACN showed clear phase separation due to differential densities in the extracts after centrifugation. The number of extraction cycles significantly increased the flavanone levels during extraction. Modulating the sample to solvent ratio increased naringin quantity in the extracts. Current research provides critical information on the role of centrifuge temperature, extraction solvent and their interactions on flavanone distribution in extracts. PMID- 21645711 TI - A new method for the determination of biogenic amines in cheese by LC with evaporative light scattering detector. AB - This paper presents a new LC method with evaporative light scattering detection (ELSD), for the separation and determination of the biogenic amines (histamine, spermidine, spermine, tyramine, putrescine and beta-phenylethylamine) which are commonly present in cheese, as their presence and relative amounts give useful information about freshness, level of maturing, quality of storage and cheese authentication. The LC-ELSD method is validated by comparison of the results with those obtained through LC-UV determination, based on a pre-column dansyl chloride derivatisation step. The obtained data demonstrate that both methods can be interchangeably used for biogenic amines determination in cheese. The new LC-ELSD method shows good precision and permits to achieve, for standard solutions, limit of detection (LOD) values ranging from 1.4 to 3.6 mg L(-1) and limit of quantitation (LOQ) values ranging from 3.6 to 9.3 mg L(-1). The whole methodology, comprehensive of the homogenization-extraction process and LC-ELSD analysis, has been applied in the analysis of a typical Calabria (Southern Italy) POD cheese, known as Caciocavallo Silano. The most aboundant amine found was histamine, followed, in decreasing order, by tyramine, spermine, putrescine, beta phenylethylamine and spermidine, for a total amount of 127 mg kg(-1). This value does not represent a possible risk for consumer health, according to the toxicity levels reported in literature and regarded as acceptable. PMID- 21645712 TI - Multicomponent analytical methodology to control phthalates, synthetic musks, fragrance allergens and preservatives in perfumes. AB - A simple, fast, robust and reliable multicomponent analytical method applicable in control laboratories with a high throughput level has been developed to analyze commercial brands of perfumes. Contents of 52 cosmetic ingredients belonging to different chemical families can be determined in a single run. Instrumental linearity, precision of the method and recovery studies in real samples showed excellent results, so that quantification by external calibration can be effectively applied. Relevant limits of detection and quantification were obtained for all the targets considered, far below the legal requirements and amply adequate for its accurate analytical control. A survey of 70 commercial perfumes and colognes has been performed, in order to verify whether these products complied with the recent changes in European legislation: regarding the maxima allowed concentrations of the ingredients and/or ingredient labelling. All samples contained some of the target ingredients. Several samples do not comply with the regulations concerning the presence of phthalates. Musks data confirmed the trend about the replacement of nitromusks by polycyclic musks; as well as the noticeable introduction of macrocyclic musks in the perfumes composition. The prohibited musk moskene has been detected in one sample in an appreciable concentration. The average number of fragrance allergens is twelve per sample; their presence must be indicated in the list of ingredients when its concentration exceeds the 0.001%, but values higher than 1% have been found in some samples. Preservatives data show that parabens, although ubiquitous in other cosmetic products, are not widely used in perfumery. In contrast, the presence of BHT is indeed widespread. The degree of compliance with the European Regulation on the labelling has been evaluated in a subset of samples, and only about the 38% of the perfumes were properly labelled for the allergens tested. PMID- 21645713 TI - A miniature and field-applicable multipumping flow analyzer for ammonium monitoring in seawater with fluorescence detection. AB - In this article, a simple, economic, and miniature flow analyzer for ammonium in seawater based on the solenoid micropumps is presented. A single reagent of sodium tetraborate, ortho-phthaldialdehyde (OPA), and sodium sulfite was used and optimized applying the modified SIMPLEX method. A special-made detection cell for fluorescence detection of the reaction product isoindol-1-sulfonat was made and combined with a commercial photomultiplier tube, a long-pass optical filter, and an UV-LED as excitation light source. A LOD down to 13 nmol/L was achieved. The fabrication and application of a miniature reaction coil heating device for reaction rate enhancement is further described. The system featured an injection frequency of 32 h(-1) at average standard deviation of 3%. PMID- 21645714 TI - Rapid characterization of dry cured ham produced following different PDOs by proton transfer reaction time of flight mass spectrometry (PTR-ToF-MS). AB - In the present study, the recently developed proton transfer reaction time of flight mass spectrometry (PTR-ToF-MS) technique was used for the rapid characterization of dry cured hams produced according to 4 of the most important Protected Designations of Origin (PDOs): an Iberian one (Dehesa de Extremadura) and three Italian ones (Prosciutto di San Daniele, Prosciutto di Parma and Prosciutto Toscano). In total, the headspace composition and respective concentration for nine Spanish and 37 Italian dry cured ham samples were analyzed by direct injection without any pre-treatment or pre-concentration. Firstly, we show that the rapid PTR-ToF-MS fingerprinting in conjunction with chemometrics (Principal Components Analysis) indicates a good separation of the dry cured ham samples according to their production process and that it is possible to set up, using data mining methods, classification models with a high success rate in cross validation. Secondly, we exploited the higher mass resolution of the new PTR-ToF-MS, as compared with standard quadrupole based versions, for the identification of the exact sum formula of the mass spectrometric peaks providing analytical information on the observed differences. The work indicates that PTR ToF-MS can be used as a rapid method for the identification of differences among dry cured hams produced following the indications of different PDOs and that it provides information on some of the major volatile compounds and their link with the implemented manufacturing practices such as rearing system, salting and curing process, manufacturing practices that seem to strongly affect the final volatile organic profile and thus the perceived quality of dry cured ham. PMID- 21645715 TI - Electromembrane extraction (EME) and HPLC determination of non-steroidal anti inflammatory drugs (NSAIDs) in wastewater samples. AB - In this paper, an electromembrane extraction (EME) combined with a HPLC procedure using diode array (DAD) and fluorescence detection (FLD) has been developed for the determination of six widely used non-steroidal anti-inflammatory drugs (NSAIDs): salicylic acid (SAC), ketorolac (KTR), ketoprofen (KTP), naproxen (NAX), diclofenac (DIC) and ibuprofen (IBU). The drugs were extracted from basic aqueous sample solutions, through a supported liquid membrane (SLM) consisting of 1-octanol impregnated in the walls of a S6/2 Accurel((r)) polypropylene hollow fiber, and into a basic aqueous acceptor solution resent inside the lumen of the hollow fiber with a potential difference of 10 V applied over the SLM. Extractions that were carried out in 10 min using a potential of 10 V from pH 12 NaOH aqueous solutions shown concentration enrichments factors of 28-49 in a pH 12 NaOH aqueous acceptor solution. The proposed method was successfully applied to urban wastewaters. Excellent selectivity was demonstrated as no interfering peaks were detected. The procedure allows very low detection and quantitation limits of 0.0009-9.0 and 0.003-11.1 MUg L(-1), respectively. PMID- 21645716 TI - Specific O2- generation in corona discharge for ion mobility spectrometry. AB - This study deals with O(2)(-) generation in corona discharge (CD) in point to plane geometry for single flow ion mobility spectrometry (IMS) with gas outlet located behind the ionization source. We have designed CD of special geometry in order to achieve the high O(2)(-) yield. Using this ion source we have achieved in zero air conditions that up to 74% all negative ions were O(2)(-) or O(2)( )(H(2)O). It has been demonstrated that the non-electronegative nitrogen positively influences the efficiency of O(2)(-) generation in O(2)/N(2) mixtures. The reduced ion mobility of 2.27 cm(2)V(-1)s(-1) has been measured for O(2)( )/O(2)(-)(H(2)O) ions in zero air. Additional ions detected in zero air (less than 200 ppb CO(2)) using the mass spectrometric and IMS technique were, NO(2)( ), N(2)O(2)(-) (2.37 cm(2)V(-1)s(-1)), NO(3)(-), N(2)O(3)(-) and N(2)O(3)( )(H(2)O). The CO(3)(-) and CO(4)(-) ions have been detected after the introduction of 5 ppm CO(2) into zero air. PMID- 21645717 TI - Determination of airborne carbonyls via pentafluorophenylhydrazine derivatisation by GC-MS and its comparison with HPLC method. AB - The classical analytical method for gaseous carbonyl measurements based on solid sorbent coated with 2,4-dinitrophenylhydrazine (DNPH) and analysis by HPLC/UV suffers from limited resolution of carbonyls with similar molecular structures and high molecular weights. In this paper, we report the development of a sensitive and reliable analytical method for simultaneous determination of 21 airborne carbonyls within the C(1)-C(9) range. Carbonyls were collected on a sampling tube filled with 100mg Tenax TA (60-80 mesh) sorbent coated with 1 MUmol pentafluorophenyl hydrazine (PFPH), followed by solvent desorption and analysis by gas chromatography (GC)/mass spectrometry (MS). Common carbonyl gases including formaldehyde, acetaldehyde, butyraldehyde, hexaldehyde and benzaldehyde at ppbv levels were collected with efficiency greater than 90% onto sampling tubes at a flow rate of 100 mL min(-1). The limits of detection (LODs, signal/noise=3) of the tested carbonyls were in the range of 0.08-0.20 ppbv for a sampled volume of 24.0 L. These limits are less than or comparable with those that can be obtained using the DNPH-HPLC method. The method has been field-tested both in ambient air of York and in diluted cigarette smoke. Comparing field tests with the classical DNPH-HPLC method, good agreement was displayed between the two methods for the same carbonyls, but with more carbonyl species detected by the PFPH-GC/MS method. The PFPH-GC/MS method provides better molecular separation for carbonyls with similar structures, is highly sensitivity and gives confirmation of identification by structures when detected using MS. PMID- 21645718 TI - Direct determination of manganese in produced waters from petroleum exploration by Electrothermal Atomic Absorption Spectrometry using Ir-W as permanent modifier. AB - This present work reports the development and evaluation of a method for the direct determination of manganese in waters extracted during petroleum exploitation by Electrothermal Atomic Absorption Spectrometry (ET AAS) using Ir-W as permanent modifier. These waters, usually called produced waters, contain a wide range of organic and inorganic substances and are characterized by their high salinity. In order to achieve suitable experimental conditions for the method application, studies about the effect of operational variables (chemical modifier, pyrolysis and atomization temperatures) were performed, as well as the establishment of convenient calibration strategy. The best results were verified when the temperatures of pyrolysis and atomization were 1000 degrees C and 2300 degrees C, respectively, and using Ir-W as permanent modifier. The results showed that manganese can be determined by the standard addition method or employing external calibration with standard solutions prepared in the same salinity of the samples (with NaCl). Three real samples with salinities varying between 74 and 840/00 were successfully analyzed by the developed procedure. The limits of detection and quantification were 0.24 and 0.80 MUg L(-1), respectively, in purified water, and 0.34 and 1.1 MUg L(-1), respectively, in 0.4 mol L(-1) NaCl medium (approximately 230/00 salinity). PMID- 21645719 TI - Quantitative determination by temperature dependent near-infrared spectra: a further study. AB - Quantitative spectra-temperature relationship (QSTR) between near-infrared (NIR) spectra and temperature has been studied in our previous work (Talanta, 2010, 82, 1017-1021). In this study, applicability of the QSTR model for quantitative determination is further studied using the spectra of aqueous ethanol samples in the temperature range of 31-40 degrees C and the concentration range of 1-99%. The results show that QSTR model can be built by using the spectra in a small temperature range and the quantitative analysis can be achieved by only two spectra at different temperatures. Moreover, calibration curves for different concentration ranges (1-5%, 20-70%, 95-99%, v/v) are investigated by using linear and nonlinear curve fitting, respectively. Both of the linear and nonlinear curves are found to be applicable within these concentration ranges. Therefore, the temperature dependent NIR spectra may provide a new way for quantitative determination and may have high potential in bio-fluids analysis or industrial practices. PMID- 21645720 TI - Electrochemically monitoring the binding of concanavalin A and ovalbumin. AB - To evaluate protein-protein interactions, a new voltammetric method was developed using a protein labeled with an electroactive compound. Concanavalin A (ConA), which is a lectin, recognizes alpha-mannose residues. Because the ConA was to be bound to ovalbumin (OVA), which has a high-mannose sugar chain, ConA labeled with daunomycin was prepared as the probe to monitor the binding. The binding to OVA was caused by the label modification of the ConA. As a result, the electrode response of the labeled ConA decreased as the OVA concentration increased. The electrode response of the labeled ConA was linearly over the range of 1.5*10(-10) and 1.5*10(-9)M OVA. The relative standard deviation of 1.5*10(-8)M labeled ConA and 1.5*10(-10)M OVA was 6.9% (n=5). The labeled ConA-OVA binding could then be conveniently monitored based on the change in response. In contrast, interactions between the labeled ConA and a protein with no specific sugar chain also were investigated. Incubation scarcely influenced the peak current of the labeled ConA. When several concentrations of OVA were added to a serum, good recovery determined it. Consequently, this method could be applied to the measurement of protein-protein interactions. PMID- 21645721 TI - Determination of the oxidation stability of biodiesel and oils by spectrofluorimetry and multivariate calibration. AB - Oxidation stability is an important quality parameter for biodiesel. In general, the methods used to evaluate the oxidation stability of oils and biodiesels are time-consuming. This work reports the use of spectrofluorimetry, a fast analytical technique, associated with multivariate data analysis as a powerful analytical tool to prediction of the oxidation stability. The prediction of the oxidation stability showed a good agreement with the results obtained by the EN14112 reference method Rancimat. The models presented high correlation (0.99276 and 0.97951) between real and predicted values. The R(2) values of 0.98557 and 0.95943 indicated the accuracy of the models to predict the oxidation stability of soy oil and soy biodiesel, respectively. The residual distribution does not follow a trend with respect to the predicted variables indicating the good quality of the fits. PMID- 21645722 TI - Evaluation of laser induced breakdown spectroscopy for multielemental determination in soils under sewage sludge application. AB - Laser induced breakdown spectroscopy (LIBS) is an atomic emission spectroscopy technique for simple, direct and clean analysis, with great application potential in environmental sustainability studies. In a single LIBS spectrum it is possible to obtain qualitative information on the sample composition. However, quantitative analysis requires a reliable model for analytical calibration. Multilayer perceptron (MLP), an artificial neural network, is a multivariate technique that is capable of learning to recognize features from examples. Therefore MLP can be used as a calibration model for analytical determinations. Accordingly, the present study proposes to evaluate the traditional linear fit and MLP models for LIBS calibration, in order to attain a quantitative multielemental method for contaminant determination in soil under sewage sludge application. Two sets of samples, both composed of two kinds of soils were used for calibration and validation, respectively. The analyte concentrations in these samples, used as reference, were determined by a reference analytical method using inductively coupled plasma optical emission spectrometry (ICP OES). The LIBS-MLP was compared to a LIBS-linear fit method. The values determined by LIBS MLP showed lower prediction errors, correlation above 98% with values determined by ICP OES, higher accuracy and precision, lower limits of detection and great application potential in the analysis of different kinds of soils. PMID- 21645723 TI - Voltammetric behavior of benzo[a]pyrene at boron-doped diamond electrode: a study of its determination by adsorptive transfer stripping voltammetry based on the enhancement effect of anionic surfactant, sodium dodecylsulfate. AB - Benzo[a]pyrene (BaP), a member of the polycyclic aromatic hydrocarbon (PAH) class, is one of the most potent PAH carcinogens. The electrochemical oxidation of BaP was first studied by cyclic voltammetry at the boron-doped diamond electrode in non-aqueous solvent (dimethylsulphoxide with lithium perchlorate). The compound was irreversibly oxidized in a single step at high positive potential, resulting in the well-resolved formation of a couple with a reduction and re-oxidation wave at much lower potentials. Special attention was given to the use of adsorptive stripping voltammetry together with a medium exchange procedure in aqueous and aqueous/surfactant solutions over the pH range of 2.0 8.0. The technique in aqueous solutions had little value in practice because of too small oxidation peak current. This problem was solved when surfactants were added into the sample solution, by which the oxidation peak currents of BaP were found enhanced dramatically. The employed surfactants were sodium dodecylsulfate (anionic, SDS), cetyltrimethylammonium bromide (cationic, CTAB) and Tween 80 (non ionic). Using square-wave stripping mode, the compound yielded a well-defined voltammetric response in Britton-Robinson buffer, pH 2.0 containing 2.5*10(-4)M SDS at +1.07 V (vs. Ag/AgCl) (after 120s accumulation at +0.10 V). The process could be used to determine BaP in the concentration range of 16-200 nM (4.04 50.46 ng mL(-1)), with a detection limit of 2.86 nM (0.72 ng mL(-1)). This method was also applied to determine BaP in model water sample prepared by adding its different concentrations into tap water. PMID- 21645724 TI - Improved solid-phase extraction/micellar procedure for the derivatization/preconcentration of benzaldehyde and methyl derivatives from water samples. AB - A simple, rapid and sensitive method has been developed for the determination of aromatic low-molecular mass aldehydes (LMMAs) such as benzaldehyde (BA) and methyl derivatives in water samples through the use of liquid chromatography diode array detection (LC-DAD). The method is based on the continuous in situ derivatization of the aldehydes with 2,4-dinitrophenylhydrazine (DNPH) on a LiChrolut EN solid-phase extraction (SPE) column in the presence of sodium dodecyl sulfate (SDS) micelles. After elution, hydrazones were successfully separated on a RP-C(18) column using a linear gradient mobile phase of acetonitrile (ACN)-water at 75-95% ACN for 10 min. Linearity was established over the concentration range 0.4-200 MUg L(-1) and limits of detection (LODs) from 120 to 200 ng L(-1); the inter-day precision expressed as the relative standard deviation (RSD) of the aldehydes ranged from 3.0% to 3.5%. The method was successfully applied to the analysis of aromatic and aliphatic LMMAs in water samples with average recoveries ranging between 93.6% and 99.5%. The proposed method surpasses other chromatographic alternatives in terms of LODs, sample requirements for analysis and cost. PMID- 21645725 TI - Assessing the effect of oxygen and microbial inhibitors to optimize ferricyanide mediated BOD assay. AB - Methods for short-term BOD analysis (BOD(st)) based on ferricyanide mediator reduction have succeeded in overcoming some problems associated with the standard BOD test analysis (BOD(5)) such as long-term incubations (5 days), the need to dilute samples and low reproducibility. Here we present a bioassay where a Klebsiella pneumoniae environmental strain successfully reduces ferricyanide without de-aeration of the samples with linear BOD(5) ranges between 30 and 500 mg L(-1) or 30 and 200 mg L(-1), using glucose-glutamic acid solution (GGA) or OECD standards respectively. We further propose a new assay termination solution that allows higher reproducibility and standardization of the cell-based assay, employing formaldehyde (22.7 g L(-1)) or other compounds in order to stop ferricyanide reduction without affecting the amperometric detection and therefore replace the centrifugation step normally used to stop microbial-driven reactions in ferricyanide-mediated bioassays. These improvements led to an accurate determination of real municipal wastewater samples. PMID- 21645726 TI - Sensing properties of polyethylenimine coated carbon nanotubes in oxidized oil. AB - Chemical detection is still a continuous challenge when it comes to designing single-walled carbon nanotube (SWCNT) sensors with high selectivity, especially in complex chemical environments. A perfect example of such an environment would be in thermally oxidized soybean oil. At elevated temperatures, oil oxidizes through a series of chemical reactions that results in the formation of monoacylglycerols, diacylglycerols, oxidized triacylglycerols, dimers, trimers, polymers, free fatty acids, ketones, aldehydes, alcohols, esters, and other minor products. In order to detect the rancidity of oxidized soybean oil, carbon nanotube chemiresistor sensors have been coated with polyethylenimine (PEI) to enhance the sensitivity and selectivity. PEI functionalized SWCNTs are known to have a high selectivity towards strong electron withdrawing molecules. The sensors were very responsive to different oil oxidation levels and furthermore, displayed a rapid recovery of more than 90% in ambient air without the need of heating or UV exposure. PMID- 21645727 TI - 3-Aminopropyltriethoxysilane-functionalized manganese doped ZnS quantum dots for room-temperature phosphorescence sensing ultratrace 2,4,6-trinitrotoluene in aqueous solution. AB - New strategies for silica coating of inorganic nanoparticles became a research hotspot for enhancing the mechanical stability of colloidal particles and protecting colloidal particles against oxidation and agglomeration, and so on. In this paper, 3-aminopropyltriethoxysilane (APTES)-functionalized Mn doped (AF MnD) ZnS QDs was prepared to be firsyly through the use of silane coupling agents to form an active layer of silica, then sol-gel reaction of TEOS co-deposited with APTES on the surface of resultant active layer of silica. The emitted long lifetime room-temperature phosphorescence (RTP) of the resultant nanomaterials allows an appropriate delay time so that any fluorescent emission and scattering light can be easily avoided. The APTES anchored on the layer of silica can bind 2,4,6-trinitrotoluene (TNT) species to form TNT anion through acid-base pairing interaction, the TNT anion species may increase the charge-transfer pathways from the nanocrystals to nitroaromatic analytes, therefore further enhance the quenching efficiency of RTP. Moreover, APTES as capped reagents can enlarge the spectral sensitivity and enhance RTP response of nanocrystals to the electron deficient nitroaromatic and nitrophenol species. Meanwhile, AF MnD ZnS QDs also exhibited a highly selective response toward TNT analyte through significant color change and quenching of (4)T(1) to (6)A(1) transition emission. This AF MnD ZnS QDs based sensor showed a very good linearity in the range of 0.05-1.8MUM with detection limit down to 50 nM (quenching percentage of phosphorescence intensity of 8%) and RSD of 3.5% (n=5). The reported QDs-based chemosensors here open up a promising prospect for the sensitive and convenient sensing of TNT explosive. PMID- 21645728 TI - Gold nanorod-catalyzed luminol chemiluminescence and its selective determination of glutathione in the cell extracts of Saccharomyces cerevisiae. AB - In this study, gold nanorods were firstly found to exhibit a tremendously higher catalytic activity towards luminol chemiluminescence (CL) than spherical gold nanoparticles. More importantly, ultra-trace aminothiols can cause a great CL decrease in the gold nanorod-catalyzed luminol system by the formation of Au-S covalent bonds on the ends of gold nanorods. Aminothiols can occupy the active sites of gold nanorods, and further interrupt the generation of the active oxygen intermediates. Other biomolecules including 19 standard amino acids, alcohols, organic acids and saccharides have no effect on gold nanorod-catalyzed luminol CL signals. Moreover, in order to evaluate the applicability and reliability of the proposed method, it was applied to the determination of glutathione in the cell extracts of Saccharomyces cerevisiae. Good agreements were obtained for the determination of glutathione in the cell extracts of S. cerevisiae between the present approach and a standard Alloxan method. The recoveries of glutathione were found to fall in the range between 96 and 105%. The calibration curve for glutathione was found to be linear from 0.05 to 100 nM, and the detection limit (S/N=3) was 0.01 nM. The relative standard deviation (RSD) for five repeated measurements of 5.0 nM glutathione was 2.1%. PMID- 21645729 TI - Preparation of poly(vinyltetrazole) chain-grafted poly(glycidymethacrylate-co ethylenedimethacrylate) beads by surface-initiated atom transfer radical polymerization for the use in weak cation exchange and hydrophilic interaction chromatography. AB - A novel stationary phase for weak cation exchange (WCX) and hydrophilic interaction chromatography (HILIC) was prepared with surface-initiated atom transfer radical polymerization (SI-ATRP). Vinyltetrazole was grafted onto the surface of the beads in water medium with the polyglycidylmethacrylate beads (P(GMA/EDMA)) previously modified with 2-bromoisobutryl bromide as the macromolecule initiators and CuCl as catalyst. The poly(vinyltetrazole)-grafted beads obtained with different atom transfer radical polymerization (ATRP) formulations were tried as chromatographic packings in ion-exchange chromatography. The results showed that the prepared columns could separate the tested proteins with high efficiency and high capacity, and the retention time of protein had a positive relationship with increasing the chain lengths of the grafted poly(vinyltetrazole) (PVT). The prepared column was also found to be able to separate nucleosides by hydrophilic interaction chromatographic mode. PMID- 21645730 TI - Determination of bisphenol A and naphthols in river water samples by capillary zone electrophoresis after cloud point extraction. AB - As a first attempt, cloud point extraction (CPE) was developed to preconcentrate bisphenol A (BPA), alpha-naphthol and beta-naphthol prior to performing capillary zone electrophoresis (CZE) analysis. The parameters influencing the CPE efficiency, such as Triton X-114 concentrations, pH value, extraction time and temperature were systematically evaluated. After diluting with acetonitrile, the surfactant-rich phase of CPE can be injected directly into the CE instrument. The CZE baseline separation was achieved with running buffer (pH 9.5) composed of 50mM sodium tetraborate in 30% (v/v) methanol, and an applied voltage of 25 kV. Under the optimized CPE and CZE conditions, an preconcentration factor of 50 times could be obtained and the limit of quantification for the three analytes were found to be 1.67 MUg L(-1), 0.80 MUg L(-1) and 0.67 MUg L(-1) for BPA, alpha naphthol and beta-naphthol, respectively. The proposed methods have shown to be a green, rapid and effective approach for determination of three analytes present in river water samples. PMID- 21645731 TI - Layer-by-layer assembly of electroactive dye/inorganic matrix film and its application as sensor for ascorbic acid. AB - A novel inorganic-organic composite ultrathin film was fabricated by layer-by layer assembly of naphthol green B (NGB) and layered double hydroxides (LDHs) nanoplatelets, which shows remarkable electrocatalytic behavior for oxidation of ascorbic acid. LDHs nanoplatelets were prepared using a method involving separate nucleation and aging steps (particle size: 25+/-5 nm; aspect ratio: 2-4) and used as building blocks for alternate deposition with NGB on indium tin oxide (ITO) substrates. UV-vis absorption spectroscopy and XRD display regular and uniform growth of the NGB/LDHs ultrathin film with extremely c-orientation of LDHs nanoplatelets (ab plane of microcrystals parallel to substrates). A continuous and uniform surface morphology was observed by SEM and AFM image. The film modified electrode displays a couple of well-defined reversible redox peaks attributed to Fe(2+)/Fe(3+) in NGB (DeltaE(p)=68 mV and I(a)/I(c)=1.1). Moreover, the modified electrode shows a high electrocatalytic activity towards ascorbic acid in the range 1.2-55.2 MUM with a detection limit of 0.51 MUM (S/N=3). The Michaelis-Menten constant was calculated to be K(M)(app)=67.5 MUM. PMID- 21645732 TI - Large-scale assignment of N-glycosylation sites using complementary enzymatic deglycosylation. AB - Endoglycosidase is a class of glycosidases that specifically cleaves the glycosidic bond between two proximal residues of GlcNAc in the pentasaccharide core of N-glycan, leaving the innermost GlcNAc still attached to its parent protein, which provides a different diagnostic maker for N-glycosylation site assignment. This study aims to validate the use of endoglycosidase for high throughput N-glycosylation analysis. An endoglycosidase of Endo H and the conventional PNGase F were employed, with a similar accessible procedure, for large-scale assignment of N-glycosylation sites and then N-glycoproteome for rat liver tissue. ConA affinity chromatography was used to enrich selectively high mannose and hybrid glycopeptides before enzymatic deglycosylation. As a result, a total of 1063 unique N-glycosites were identified by nano liquid chromatography tandem mass spectrometry, of which 53.0% were unknown in the Swiss-Prot database and 47.1% could be assigned only by either of the methods, confirmed the possibility of large-scale glycoproteomics by use of endoglycosidase. In addition, 11 glycosites were assigned with core-fucosylation by Endo H. A comparison between the two enzymatic deglycosylation methods was also investigated. Briefly, Endo H provides a more confident assignment but a smaller dataset compared with PNGase F, showing the complementary nature of the two N glycosite assignment methods. PMID- 21645733 TI - Direct simultaneous determination of Co, Cu, Fe, Ni and V in pore waters by means of adsorptive cathodic stripping voltammetry with mixed ligands. AB - An analytical procedure is proposed for the direct simultaneous determination in a single scan of Co, Cu, Fe, Ni and V in sediment pore waters by means of adsorptive cathodic stripping voltammetry (ACSV) with mixed ligands (DMG and catechol). Optimum conditions for the determination of these five elements were studied. Detection limits of the technique depended upon the reproducibility of the procedure blank, and were found to be 0.04 nM Co, 0.09 nM Cu, 1.29 nM Fe, 0.46 nM Ni and 2,52 nMV making the method suitable for the direct simultaneous determination of these five metals in pore waters, estuarine waters and probably coastal waters. PMID- 21645734 TI - An electrochemical immunosensor based on covalent immobilization of okadaic acid onto screen printed carbon electrode via diazotization-coupling reaction. AB - In this work, an electrochemical method based on the diazonium-coupling reaction mechanism for the immobilization of okadaic acid (OA) on screen printed carbon electrode was developed. At first, 4-carboxyphenyl film was grafted by electrochemical reduction of 4-carboxyphenyl diazonium salt, followed by terminal carboxylic group activation by N-hydroxysuccinimide (NHS), N-(3 dimethylaminopropyle)-N'-ethyle-carbodiimide hydrochloride (EDC). Hexamethyldiamine was then covalently bound by one of its terminal amine group to the activated carboxylic group. The carboxyl group of okadaic acid was activated by EDC/NHS and then conjugated to the second terminal amine group on other side of the hexamethyldiamine through amide bond formation. After immobilization of OA, an indirect competitive immunoassay format was employed to detect OA. The immunosensor obtained using this novel approach allowed detection limit of 1.44 ng/L of OA, and was also validated with certified reference mussel samples. PMID- 21645735 TI - Comparison of biosensor platforms for surface plasmon resonance based detection of paralytic shellfish toxins. AB - Paralytic shellfish poisoning (PSP) toxins are produced by certain marine dinoflagellates and may accumulate in bivalve molluscs through filter feeding. The Mouse Bioassay (MBA) is the internationally recognised reference method of analysis, but it is prone to technical difficulties and regarded with increasing disapproval due to ethical reasons. As such, alternative methods are required. A rapid surface plasmon resonance (SPR) biosensor inhibition assay was developed to detect PSP toxins in shellfish by employing a saxitoxin polyclonal antibody (R895). Using an assay developed for and validated on the Biacore Q biosensor system, this project focused on transferring the assay to a high-throughput, Biacore T100 biosensor in another laboratory. This was achieved using a prototype PSP toxin kit and recommended assay parameters based on the Biacore Q method. A monoclonal antibody (GT13A) was also assessed. Even though these two instruments are based on SPR principles, they vary widely in their mode of operation including differences in the integrated MU-fluidic cartridges, autosampler system, and sensor chip compatibilities. Shellfish samples (n=60), extracted using a simple, rapid procedure, were analysed using each platform, and results were compared to AOAC high performance liquid chromatography (HPLC) and MBA methods. The overall agreement, based on statistical 2*2 comparison tables, between each method ranged from 85% to 94.4% using R895 and 77.8% to 100% using GT13A. The results demonstrated that the antibody based assays with high sensitivity and broad specificity to PSP toxins can be applied to different biosensor platforms. PMID- 21645736 TI - Improvement of the analysis of the biochemical oxygen demand (BOD) of Mediterranean seawater by seeding control. AB - Biochemical oxygen demand (BOD) is a useful parameter for assessing the biodegradability of dissolved organic matter in water. At the same time, this parameter is used to evaluate the efficiency with which certain processes remove biodegradable natural organic matter (NOM). However, the values of BOD in seawater are very low (around 2 mgO(2)L(-1)) and the methods used for its analysis are poorly developed. The increasing attention given to seawater desalination in the Mediterranean environment, and related phenomena such as reverse osmosis membrane biofouling, have stimulated interest in seawater BOD close to the Spanish coast. In this study the BOD analysis protocol was refined by introduction of a new step in which a critical quantity of autochthonous microorganisms, measured as adenosine triphosphate, is added. For the samples analyzed, this improvement allowed us to obtain reliable and replicable BOD measurements, standardized with solutions of glucose-glutamic acid and acetate. After 7 days of analysis duration, more than 80% of ultimate BOD is achieved, which in the case of easily biodegradable compounds represents nearly a 60% of the theoretical oxygen demand. BOD(7) obtained from the Mediterranean Sea found to be 2.0+/-0.3 mgO(2)L(-1) but this value decreased with seawater storage time due to the rapid consumption of labile compounds. No significant differences were found between two samples points located on the Spanish coast, since their organic matter content was similar. Finally, the determination of seawater BOD without the use of inoculum may lead to an underestimation of BOD. PMID- 21645737 TI - Selective colorimetric sensing of cysteine in aqueous solutions using silver nanoparticles in the presence of Cr3+. AB - We here in report an extensive study on the development of a highly facile, selective and sensitive colorimetric probe for cysteine detection using silver nanoparticles (Ag NPs). The efficacy of the process relies upon the surface plasmon resonance properties of Ag NPs and the interaction of Ag-cysteine complex with chromium ions (Cr(3+)) in a ratio of 2:1. In the presence of Cr(3+), cysteine was able to induce the aggregation of Ag NPs thereby resulting in a change in yellow colour of the Ag colloid to purple. The reported probe has a limit of detection down to 1 nM which is to the best of our knowledge the lowest ever reported for the colorimetric detection of cysteine. Furthermore, a remarkable feature of this method is that it involves a simple technique exhibiting high selectivity to cysteine over other tested amino acids. PMID- 21645738 TI - A dispersive liquid-liquid microextraction procedure for determination of boron in water after ultrasound-assisted conversion to tetrafluoroborate. AB - A novel, simple and green procedure is presented for the determination of boron. The method is based on ultrasound-assisted conversion of boron to tetrafluoroborate anion and the formation of an ion pair between BF(4)(-) and Astra Phloxine reagent (R), followed by dispersive liquid-liquid microextraction of the ion pair formed and subsequent UV-vis spectrophotometric detection. The conversion of boron to tetrafluoroborate anion is performed in an acidic medium of 0.9 mol L(-1) H(2)SO(4) in the presence of 0.1 mol L(-1)F(-) by means of 10 min of ultrasonication. The extraction of the ion pair formed between BF(4)(-) and R (1*10(-4)mol L(-1)R) is carried out by dispersive liquid-liquid microextraction using 0.5 mL of amyl acetate (as extraction solvent), tetrachloromethane (as auxiliary solvent) and acetonitrile (as dispersive solvent) in a ratio of 1:1:2. The absorbance of the coloured extracts obeys Beer's law in the range 0.22-18.7 mg L(-1) of B(III) at 553 nm wavelength. The limit of detection calculated from a blank test (n=10) based on 3s is 0.015 mg L( 1) of B(III). The method was applied to the determination of boron in mineral waters. PMID- 21645739 TI - Catalytic-adsorptive stripping voltammetric determination of ultra-trace iridium(III). Application to fresh- and sea-water. AB - An extremely sensitive stripping voltammetric procedure for ultra-trace determination of iridium(III) is reported. The method is based on the interfacial accumulation of the iridium(III)-CTAB complex onto the glassy carbon electrode, followed by the catalytic reduction of the adsorbed complex in the presence of bromate. 0.3 mol L(-1) acetate buffer pH 4.7+6.9*10(-2) mol L(-1) NaBrO(3)+2.7*10(-5)mol L(-1) cetyltrimethylammonium bromide (CTAB)+0.2 mol L(-1) KCl was employed as the supporting electrolyte. The analytical procedure was verified by the analysis of the standard reference materials: Sea Water BCR-CRM 403 and Fresh Water NIST-SRM 1643d. The accuracy, expressed as relative error e%, was satisfactory, being lower than 6%, while precision as repeatability, expressed as relative standard deviation s(r)%, was generally lower than 5%. The limit of detection was of the order of 2-3 ng L(-1). Once set up on the standard reference materials, the analytical procedure was transferred and applied to superficial water sampled in proximity to superhighway and in the Po river mouth area. PMID- 21645740 TI - Solid phase extractive preconcentration coupled to gas chromatography-atomic emission detection for the determination of chlorophenols in water samples. AB - Solid-phase extraction (SPE) followed by derivatization and gas chromatography atomic emission detection (GC-AED) was evaluated for the determination of five chlorophenols (CPs) in water samples. The derivatization was based on the esterification of phenolic compounds with ferrocenecarboxylic acid. The determination of the derivatized phenols was performed by GC-AED in the iron selective detection mode at 302 nm. The described method was tested on spiked water samples.The overall method gave detection limits of 1.6-3.7 ng L(-1) and recoveries of 90.9-104.5% for the examined mono- to trichlorophenols in 10 mL water samples. The CPs extracted from a 10 mL water sample with SPE were concentrated into 100 MUL of organic solvent, a preconcentration factor of 100. The method was applied to lake and tap water samples, and CP contents between 6 and 51 ng L(-1) in lake water and between below the detection limit and 8 ng L( 1) in tap water were found for different CPs. The method is quick, simple and gives excellent recoveries, limits of detection and standard deviations. PMID- 21645741 TI - Confocal micro-X-ray fluorescence analysis as a new tool for the non-destructive study of the elemental distributions in pharmaceutical tablets. AB - Chemical imaging studies of pharmaceutical tablets are currently an important emerging field in the pharmaceutical industry. Finding the distribution of the different compounds inside the tablet is an important issue for production quality control but also for counterfeit detection. Most of the currently used techniques are limited to the study of the surface of the compacts, whereas the study of the bulk requires a time-consuming sample preparation. In this paper, we present the use of 3D micro-X-ray fluorescence analysis (3D MUXRF) for the non destructive study of pharmaceutical tablets. Based on two different examples, it was shown that it was possible to measure the distribution of several inorganic elements (Zn, Fe, Ti, Mn, Cu) from the surface to a depth of several hundred microns under the surface. The X-ray absorption, depending on both matrix composition and energy, is one of the most critical factors of this analytical method while performing depth profiling or mapping. Therefore, an original method to correct the absorption, in order to accurately measure the true elemental distribution, was proposed. Moreover, by using the presence of titanium dioxide in a pharmaceutical coating, we proved that this technique is also suited to the non-destructive measurement of coating thickness. PMID- 21645742 TI - Melamine detection by mid- and near-infrared (MIR/NIR) spectroscopy: a quick and sensitive method for dairy products analysis including liquid milk, infant formula, and milk powder. AB - Melamine (2,4,6-triamino-1,3,5-triazine) is a nitrogen-rich chemical implicated in the pet and human food recalls and in the global food safety scares involving milk products. Due to the serious health concerns associated with melamine consumption and the extensive scope of affected products, rapid and sensitive methods to detect melamine's presence are essential. We propose the use of spectroscopy data-produced by near-infrared (near-IR/NIR) and mid-infrared (mid IR/MIR) spectroscopies, in particular-for melamine detection in complex dairy matrixes. None of the up-to-date reported IR-based methods for melamine detection has unambiguously shown its wide applicability to different dairy products as well as limit of detection (LOD) below 1 ppm on independent sample set. It was found that infrared spectroscopy is an effective tool to detect melamine in dairy products, such as infant formula, milk powder, or liquid milk. ALOD below 1 ppm (0.76+/-0.11 ppm) can be reached if a correct spectrum preprocessing (pretreatment) technique and a correct multivariate (MDA) algorithm-partial least squares regression (PLS), polynomial PLS (Poly-PLS), artificial neural network (ANN), support vector regression (SVR), or least squares support vector machine (LS-SVM)-are used for spectrum analysis. The relationship between MIR/NIR spectrum of milk products and melamine content is nonlinear. Thus, nonlinear regression methods are needed to correctly predict the triazine-derivative content of milk products. It can be concluded that mid- and near-infrared spectroscopy can be regarded as a quick, sensitive, robust, and low-cost method for liquid milk, infant formula, and milk powder analysis. PMID- 21645743 TI - Rapid honey characterization and botanical classification by an electronic tongue. AB - In this paper a commercial electronic tongue (alphaAstree, Alpha M.O.S.) was applied for botanical classification and physicochemical characterization of honey samples. The electronic tongue was comprised of seven potentiometric sensors coupled with an Ag/AgCl reference electrode. Botanical classification was performed by PCA, CCA and ANN modeling on 12 samples of acacia, chestnut and honeydew honey. The physicochemical characterization of honey was obtained by ANN modeling and the parameters included were electrical conductivity, acidity, water content, invert sugar and total sugar. The initial reference values for the physicochemical parameters observed were determined by traditional methods. Botanical classification of honey samples obtained by ANN was 100% accurate while the highest correlation between observed and predicted values was obtained for electrical conductivity (0.999), followed by acidity (0.997), water content (0.994), invert sugar content (0.988) and total sugar content (0.979). All developed ANN models for rapid honey characterization and botanical classification performed excellently showing the potential of the electronic tongue as a tool in rapid honey analysis and characterization. The advantage of using such a technique is a simple sample preparation procedure, there are no chemicals involved and there are no additional costs except the initial measurements required for ANN model development. PMID- 21645744 TI - Stabilization of sensing performance for mixed-potential-type zirconia-based hydrocarbon sensor. AB - The recently reported sensing characteristics of the mixed-potential-type yttria stabilized zirconia (YSZ)-based hydrocarbon (HC) sensor attached with ZnCr(2)O(4) sensing electrode (SE) were found to be changed after the 10-day operation at 550 degrees C under the wet condition (5 vol.% water vapor). To improve the stability of the present sensor, the several modifications of the SE material by adding YSZ powder were examined. As a result, the sensor using the laminated (ZnCr(2)O(4)/YSZ)-SE gave the stable electromotive force (emf) response against 100 ppm C(3)H(6) at 550 degrees C for about one month examined. Based on the scanning electron microscopy (SEM) observation and the AC complex-impedance measurements, it was concluded that the stable behavior of the sensor using the laminated (ZnCr(2)O(4)/YSZ)-SE was provided by the stabilization of the interface between ZnCr(2)O(4) grains and YSZ particles. The fabricated sensor exhibited the linear dependence of sensitivity on the logarithm of either C(3)H(6) concentration (in the range of 20-800 ppm) or mixtures of various hydrocarbons (HCs) (in the range of 90-2600 ppmC). In addition, the emf response was not altered by the change of O(2) (2-20 vol.%), H(2)O (0-10.8 vol.%) and CO(2) (0-20 vol.%) concentrations, and no interference of other gases (CO, NO, NO(2), H(2), and CH(4)) was observed. PMID- 21645745 TI - Determination of rare earth elements and other trace elements (Y, Mn, Co, Cr) in seawater using Tm addition and Mg(OH)2 co-precipitation. AB - This paper reports on a novel procedure for determining trace element abundances (REE and Y, Cr, Mn, Co) in seawater by inductively coupled plasma sector field mass spectrometry (ICP-SFMS). The procedure uses a combination of pre concentration using co-precipitation onto magnesium hydroxides and addition of thulium spike. The validity of the method was assessed onto 25 ml volumes of certified reference materials (NASS- and CASS-4) and in house seawater standard. Procedural blanks were determined by applying the same procedure to aliquots of seawater previously depleted in trace elements by successive Mg(OH)(2) co precipitations, yielding estimated contributions to the studied samples better than 1.1% for all elements, with the exception of Cr (<3.3%) and Co (up to 8%). The reproducibility of the method over the six month duration of the study was smaller than 11% RSD for all the studied elements. Results obtained for NASS-5 and CASS-4 agree well with published working values for trace elements. PMID- 21645746 TI - Improved spectrophotometric determination of paraquat in drinking waters exploiting a Multisyringe liquid core waveguide system. AB - A novel Multisyringe flow injection analysis (MSFIA) system combined with a 200 cm long pathlength liquid core waveguide (LCW) has been developed enabling for the first time the sensitive spectrophotometric determination at MUg L(-1) levels of the herbicide paraquat (Pq(2+)) in drinking waters. The proposed system is a simple, economic and fast alternative for obtaining the first evidence of paraquat pollution prior the use of more complex instrumental techniques. The proposed methodology is based on the production of a blue free radical by reaction of Pq(2+) with ascorbic acid (partially oxidized with potassium iodate) in basic medium. Limits of detection and quantification as low as 0.7 and 2.3 MUg L(-1), were obtained respectively. The working range is linear up to a concentration of 250 MUg L(-1) of Pq(2+). The injection throughput of the proposed method is 34 h(-1). The results obtained with the LCW are compared with those using a conventional 1cm flow cell. The automation of standard addition procedures has been studied and implemented for samples causing matrix effects. Finally the proposed system has been applied to the determination of paraquat in drinking water samples. PMID- 21645747 TI - Validation of a method for the analysis of nine quinolones in eggs by pressurized liquid extraction and liquid chromatography with fluorescence detection. AB - A multiresidue method for the analysis, in egg matrices, of residues of nine quinolones used in veterinary medicine, has been developed and validated according to the provisions of Council Decision 2002/657. Compounds were extracted by a pressurized liquid extraction (PLE) technique using a 1:1 mixture of acetonitrile and a phosphoric acid buffer (pH 3.0) at 70 degrees C. The obtained extract was clear enough, so that no further clean-up was necessary. Analytes were determined by liquid chromatography (LC) with fluorescence detection (FL). Two chromatographic columns were compared: a high-purity silica Inertsil C(8) column and a newly developed Kinetex C(18) core-shell technology column. Validation was carried out at four concentration levels, using both chromatographic columns. Precision in terms of reproducibility standard deviation was between 7% and 23%, and good recoveries were obtained. Decision limit (CCalpha) and detection capability (CCbeta) values obtained with the Inertsil and Kinetex columns were in the 0.2-19.8 MUg kg(-1) and 0.4-33.5 MUg kg(-1) concentration ranges, respectively. The proposed method allows residues of quinolones banned for use with laying hens to be detected and quantified in eggs. About twenty-four samples per day can be processed. PMID- 21645748 TI - Comparison of different extraction methods for the determination of statin drugs in wastewater and river water by HPLC/Q-TOF-MS. AB - Three preconcentration techniques including solid phase extraction (SPE), dispersive liquid-liquid microextraction (DLLME) and stir-bar sorptive extraction (SBSE) have been optimized and compared for the analysis of six hypolipidaemic statin drugs (atorvastatin, fluvastatin, lovastatin, pravastatin, rosuvastatin and simvastatin) in wastewater and river water samples by high performance liquid chromatography coupled to quadrupole-time-of-flight mass spectrometry (HPLC/Q-TOF MS). Parameters that affect the efficiency of the different extraction methods such as solid phase material, sample pH and elution solvent in the case of SPE; the type and volume of the extracting and dispersive solvent, pH of sample, salt addition and number of extraction steps in the case of DLLME; and the stirring time, pH of sample, sample volume and salt addition for SBSE were evaluated. SPE allowed the best recoveries for most of the analytes. Pravastatin was poorly extracted by DLLME and could not be determined. SBSE was only applicable for lovastatin and simvastatin. However, despite the limitations of having poorer recovery than SPE, DLLME and SBSE offered some advantages because they are simple, require low organic solvent volumes and present low matrix effects. DLLME required less time of analysis, and for SBSE the stir-bar was re-usable. SPE, DLLME and SBSE provided method detection limits in the range of 0.04-11.2 ng L( 1), 0.10-17.0 ng L(-1) for 0.52-2.00 ng L(-1), respectively, in real samples. To investigate and compare their applicability, SPE, DLLME and SBSE procedures were applied to the detection of statin drugs in effluent wastewater and river samples. PMID- 21645749 TI - Novel extraction supports based on immobilised aptamers: evaluation for the selective extraction of cocaine. AB - A new kind of selective sorbent based on the use of aptamers and dedicated to the selective solid phase extraction was developed. Cocaine aptamer was chosen as model aptamer to demonstrate the feasibility of this material and to provide a complete evaluation of the synthesized sorbent. The effect of different parameters such as the nature of the immobilisation support (silica, agarose), the type of immobilisation (covalent or non-covalent) and the length of the spacer arm (C(6) or C(12)) were studied. Therefore, various oligosorbents based on different immobilisation strategies were synthesized and characterised by estimating the extraction recovery and the capacity of cocaine and the binding efficiency of aptamers. Control supports without immobilised aptamers were simultaneously studied in parallel to assess the selectivity brought by the oligosorbents. The oligosorbent based on CNBr-activated sepharose showed the best performances with an extraction recovery for cocaine of 90% while 6% was obtained on the control sorbent. The high selectivity brought by the oligosorbent was then illustrated by applying the oligoextraction followed by LC/MS analysis to a post mortem blood (cocaine overdose). Results were compared to those resulting from a conventional protein precipitation procedure. The presence of co-extracted interfering compounds was strongly reduced with the treatment by oligoextraction. A limit of quantification of 0.5 ng/mL was obtained that is largely lower than the concentration found after a single intake of cocaine. PMID- 21645750 TI - Dithizone immobilized silica gel on-line preconcentration of trace copper with detection by flame atomic absorption spectrometry. AB - A novel adsorbent-silica gel bound dithizone (H(2)Dz-SG) was prepared and used as solid-phase extraction of copper from complex matrix. The H(2)Dz-SG is investigated by means of FT-IR spectra and the SEM images, demonstrating the bonding of dithizone. The H(2)Dz-SG quantitatively adsorb copper ions, and the retained copper is afterwards collected by elution of 10% (v/v) nitric acid. An on-line flow injection solid-phase extraction procedure was developed for trace copper separation and preconcentration with detection by flame atomic spectrometry. By loading 5.4 mL of sample solution, a liner range of 0.5-120 MUg L(-1), an enrichment factor of 42.6, a detection limit of 0.2 MUg L(-1) and a precision of 1.7% RSD at the 40 MUg L(-1) level (n=11) were obtained, along with a sampling frequency of 47 h(-1). The dynamic sorption capacity of H(2)Dz-SG to Cu(2+) was 0.76 mg g(-1). The accuracy of the proposed procedure was evaluated by determination of copper in reference water sample. The potential applications of the procedure for extraction of trace copper were successfully accomplished in water samples (tap, rain, snow, sea and river). The spiking recoveries within 91 107% are achieved. PMID- 21645751 TI - A novel polybenzimidazole-modified gold electrode for the analytical determination of hydrogen peroxide. AB - The imine of polybenzimidazole (PBI) is chemically oxidized by hydrogen peroxide (H(2)O(2)) in the presence of acetic acid (AcOH). Fourier transform infrared (FT IR) and X-ray photoelectron spectroscopies (XPS) showed that when the AcOH concentration remained constant, the degree of oxidation increased with increasing H(2)O(2) levels. Moreover, the imine also exhibited electrochemical redox behavior. Based on these properties, a PBI-modified Au (PBI/Au) electrode was developed as an enzyme-free H(2)O(2) sensor. At an applied potential of -0.5V vs. Ag/AgCl, the current response of the PBI/Au electrode was linear with H(2)O(2) concentration over a range from 0.075 to 1.5mM, with a sensitivity of 55.0 MUA mM(-1)cm(-2). The probe had excellent stability, with <5% variation from its initial response current after storage at 50 degrees C for 10 days. Potentially interfering species such as ascorbic or uric acid had no effect on sensitivity. Sensitivity improved dramatically when multiwalled carbon nanotubes (MWCNT) were incorporated in the probe. Under optimal conditions, the detection of H(2)O(2) using a MWCNT-PBI/Au electrode was linear from 1.56 MUM to 2.5mM, with a sensitivity of 928.6 MUA mM(-1)cm(-2). Analysis of H(2)O(2) concentrations in urine samples using a MWCNT-PBI/Au electrode produced accurate real-time results comparable to those of traditional HPLC methods. PMID- 21645752 TI - Analysis of beryllium to biomolecule binding using a metal specific fluorescent probe and competitive assay. AB - Studying metal-biomolecule interactions is critical to the elucidation of the molecular basis of the biological functions and toxicity of metals. In the present study, a competitive fluorimetric approach has been developed to measure the apparent affinity of biomolecules for Be(2+) by using a Be(2+)-specific fluorigenic probe (10-hydroxybenzo[h]quinoline-7-sulfonate, HBQS). Under physiological conditions, HBQS coordinates with Be(2+) in a molar ratio of 1:1 and results in a fluorescence shift from 580 nm for HBQS to 480 nm for the Be HBQS complex associated with significant fluorescence enhancement. When a beryllium ligand is present in the mixture of Be(2+) and HBQS, the competition of ligand against HBQS for beryllium ion binding results in dissociation and thus a fluorescence decrease of the Be-HBQS complex. By titrating ligand and monitoring the dose-dependent decrease of Be-HBQS complex fluorescence at 480 nm, the apparent affinity between ligand and Be(2+) can be derived. Applying this simple approach, the apparent affinities of various nucleotides and the iron-storage protein ferritin for beryllium ion have been determined. In particular, the apparent dissociation constant of Be(2+) and adenosine 5'-triphosphate (ATP) was also validated by an electrospray ionization mass spectrometric (ESI-MS) method. The general applicability of the proposed competition assay was further demonstrated using FluoZin-1, a zinc fluorescent indicator, in a binding study for Zn(2+) and bovine serum albumin. This newly developed competitive fluorimetric assay provides a sensitive, simple, and generic approach for affinity estimation of metal and biomolecule binding. PMID- 21645753 TI - Combined application of alpha-track and fission-track techniques for detection of plutonium particles in environmental samples prior to isotopic measurement using thermo-ionization mass spectrometry. AB - The fission track technique is a sensitive detection method for particles which contain radio-nuclides like (235)U or (239)Pu. However, when the sample is a mixture of plutonium and uranium, discrimination between uranium particles and plutonium particles is difficult using this technique. In this study, we developed a method for detecting plutonium particles in a sample mixture of plutonium and uranium particles using alpha track and fission track techniques. The specific radioactivity (Bq/g) for alpha decay of plutonium is several orders of magnitude higher than that of uranium, indicating that the formation of the alpha track due to alpha decay of uranium can be disregarded under suitable conditions. While alpha tracks in addition to fission tracks were detected in a plutonium particle, only fission tracks were detected in a uranium particle, thereby making the alpha tracks an indicator for detecting particles containing plutonium. In addition, it was confirmed that there is a linear relationship between the numbers of alpha tracks produced by plutonium particles made of plutonium certified standard material and the ion intensities of the various plutonium isotopes measured by thermo-ionization mass spectrometry. Using this correlation, the accuracy in isotope ratios, signal intensity and measurement errors is presumable from the number of alpha tracks prior to the isotope ratio measurements by thermal ionization mass spectrometry. It is expected that this method will become an effective tool for plutonium particle analysis. The particles used in this study had sizes between 0.3 and 2.0 MUm. PMID- 21645754 TI - Standardization of fluorescence excitation-emission-matrices in aquatic milieu. AB - Fluorescence excitation-emission-matrices (EEM) are a useful tool for water quality monitoring. Recent publications show the potential of the method for real time drinking water control. However, in fluorescence measurements there is still a need for standardization to make data interpretation comparable. In this work a standardization procedure based on excitation and emission correction as well as normalization and optional inner filter effect correction is presented. By measurements of humic acid and tryptophan standards with two different spectrometers (LS 50 and LS 55 by PerkinElmer) the procedure application leads to comparable fluorescence intensities with relative standard deviations (median) of 6.6-8.4% and 10.6-12.0%, respectively. These small differences are not avoidable even if all possible correction methods are implemented and constant measurement conditions are given. The used BAM kit for emission correction induced good agreement in peak shape not only for single wavelengths but also for the whole EEM. As a consequence it is necessary to use identical equipment and identical experimental conditions in order to apply this method in fields of water quality control if small changes of fluorescence intensities are relevant for data assessment. PMID- 21645755 TI - Ligandless-solidified floating organic drop microextraction method for the preconcentration of trace amount of cadmium in water samples. AB - In this article, a new ligandless solidified floating organic drop microextraction (LL-SFODME) method has been developed for preconcentration of trace amount of cadmium as a prior step to its determination by flow injection flame atomic absorption spectrometry (FI-FAAS). The methodology is based on the SFODME of cadmium with 1-dodecanol in the absence of chelating agent. Several factors affecting the microextraction efficiency, such as, pH, sodium dodecylbenzenesulfonate (SDBS) concentration, extraction time, stirring rate and temperature were investigated and optimized. Under optimized experimental conditions an enhancement factor of 205 was obtained for 100mL of sample solution. The calibration graph was linear in the range of 1.0-25.0 ng mL(-1), the limit of detection (3s) was 0.21 ng mL(-1) and the limit of quantification (10s) was 0.62 ng mL(-1). The relative standard deviation (RSD) for 10 replicate measurements of 10 ng mL(-1) cadmium was 4.7%. The developed method was successfully applied to the extraction and determination of cadmium in standard and several water samples and satisfactory results were obtained. PMID- 21645756 TI - Comparative study of the adsorption performance of an active multi-sorbent bed tube (Carbotrap, Carbopack X, Carboxen 569) and a Radiello((r)) diffusive sampler for the analysis of VOCs. AB - A simple comparison is made to evaluate the relative performance of active and passive sampling methods for the analysis of volatile organic compounds (VOCs) in ambient air. The active sampling is done through a multi-sorbent bed tube (Carbotrap, Carbopack X, Carboxen 569) created in our laboratory and the passive sampling through the Radiello((r)) diffusive sampler specified for thermal desorption (filled with Carbograph 4). Daily duplicate samples of multi-sorbent bed tubes were taken during a period of 14 days. During the same period of time, quadruplicate samples of Radiello((r)) tubes were taken during 3 days, 4 days, 7 days and 14 days. The sampling was carried out indoors during the months of February and March 2010 and outdoors during the month of July 2010 in La Canonja (Tarragona, Spain). The analysis was performed by automatic thermal desorption (ATD) coupled with capillary gas chromatography (GC)/mass spectrometry detector (MSD). The analytical performance of the two sampling approaches was evaluated by describing several quality assurance parameters. The results show that the analytical performances of the methodologies studied are quite similar. They display low limits of detection, good precision, accuracy and desorption efficiency, and low levels of breakthrough for multi-sorbent bed tubes. However, the two monitoring methods produced varying air-borne concentration data for most of the studied compounds, and the Radiello((r)) samplers generally gave higher results. Sampling rates (Q(k)) were determined experimentally, and their values were higher than those supplied by the producer. As the experimental calculation of Q(k) values is generally carried out by the suppliers in exposure chambers with only the target compounds present in the air samples, as well as in concentrations dissimilar to those found in ambient air, the use of constant settled Q(k) can lead to inaccurate results in complex samples. PMID- 21645757 TI - Liquid chromatographic determination of sitagliptin either alone or in ternary mixture with metformin and sitagliptin degradation product. AB - Two reversed-phase liquid chromatographic (RP-LC) methods have been developed for the determination of sitagliptin phosphate monohydrate (STG). The first method comprised the determination of STG alone in bulk and plasma; and in its pharmaceutical preparation. This method was based on isocratic elution of STG using a mobile phase consisting of potassium dihydrogen phosphate buffer pH (7.8) acetonitrile (70:30, v/v) at a flow rate of 1 mL min(-1) with flourometric detection. The flourometric detector was operated at 267 nm for excitation and 575 nm for emission. In the second method, the simultaneous determination of STG and metformin (MET) in the presence of sitagliptin alkaline degradation product (SDP) has been developed. In this method, the ternary mixture of STG, MET and SDP was separated using a mobile phase consisting of potassium dihydrogen phosphate buffer pH (4.6)-acetonitrile-methanol (30:50:20, v/v/v) at a flow rate of 1 mL min(-1) with UV detection at 220 nm. Chromatographic separation in the two methods was achieved on a Symmetry((r)) Waters C18 column (150 mm*4.6 mm, 5 MUm). Linearity, accuracy and precision were found to be acceptable over the concentration ranges of 0.25-200 MUg mL(-1) for STG with the first method and 5 160 MUg mL(-1), 25-800 MUg mL(-1) for STG and MET, respectively with the second method. The optimized methods were validated and proved to be specific, robust and accurate for the quality control of the cited drugs in pharmaceutical preparations. PMID- 21645758 TI - Determination of cadmium and lead in beverages after leaching from pewter cups using graphite furnace atomic absorption spectrometry. AB - Two simple methods have been developed to determine cadmium and lead in different kinds of beverages and vinegar leached from pewter cups produced in Brazil. Leaching experiments have been carried out with different solutions: beer, sugar cane spirit, red and white wine, vinegar and a 3% acetic acid solution. The solutions were kept in cups with and without solder for 24h. Lead and cadmium have been determined using graphite furnace atomic absorption spectrometry with deuterium background correction. The limits of detection were 0.05 and 1.4 MUg L( 1), and the characteristic mass was 1.0 pg and 19 pg for Cd and Pb, respectively. With the developed methods it was possible to determine accurately cadmium and lead by direct analysis in these liquids and to evaluate the leaching of these metals from pewter cups. The results presented in this work show that pewter cups are not cadmium- and lead-free; this point goes against the manufacturers' declaration that their products are lead-free. PMID- 21645759 TI - Development of a novel flow injection liquid-liquid microextraction method for the on-line separation and preconcentration for determination of zinc(II) using 5 (8-hydroxy-2-quinolinylmethyl)-2,8-dithia-5-aza-2,6-pyridinophane as a sensitive and selective fluorescent chemosensor. AB - A novel flow injection analysis (FIA) system based on liquid-liquid microextraction and fluorimetric determination was developed for the determination of traces of the Zn(2+) ion using 5-(8-hydroxy-2-quinolinylmethyl) 2,8-dithia-5-aza-2,6-pyridinophane (L) as a sensitive and selective fluorimetric sensor, with lambda(ex)=373 nm and lambda(em)=530 nm, and hexanol as the extracting organic solvent. In the designed FIA system, the phase separation takes place via gravitation forces in the absence of any segmenter. The influence of pH and ionic strength of the solution, amount of ligand, nature of counter ion, volume of organic solvent, extraction time and coil length was investigated. Under optimized experimental conditions, the calibration curve found to be liner over a concentration range of 0.025-4.53 MUg mL(-1) (R(2)=0.9951) with a limit of detection of 2.3 ng mL(-1). The enrichment factor was 45 and relative standard deviation for 7 replicate determinations was 2.43%. The method is very fast and uses low levels of organic solvents. The proposed method was applied successfully to the determination of zinc(II) in human hair, human serum and two inorganic sludge samples. PMID- 21645760 TI - Na+-sensing quantum dots for cell-based screening of intracellular Na+ concentrations ([Na+]i). AB - We have developed a Na-quantum dot (QD) nanosensor for [Na(+)]i measurements. Using this Na-QD, we determined the dynamic physiological responses of [Na(+)]i in nonexcitable human HEK-293F cells and excitable primary rat cardiac myocytes by pharmacologically manipulating the membrane permeability to Na(+), the Na-K 2Cl cotransporter, and the Na(+)/H(+) antiporter. These data suggest that the mechanisms of [Na(+)]i homeostasis can now be elucidated with this novel Na-QD nanosensor. This could have a broad impact on Na(+) channel drug discovery. PMID- 21645761 TI - Ultrasonication-assisted extraction and preconcentration of medicinal products from herb by ionic liquids. AB - Ionic liquid-based extraction of medicinal or useful compounds from plants was investigated as an alternative to supercritical fluid, cloud point and conventional organic solvent extractions. The method integrated extraction and preconcentration. Medicinal products were first extracted by an ionic liquid solution, part of which was then converted to a hydrophobic form by anion metathesis for preconcentration. The remaining soluble ionic liquid acted as a dispersive agent to enhance the efficiency of preconcentration. Protein in the extract was precipitated spontaneously without addition of further solvents. Ultrasonication assisted this method for extraction and preconcentration of cryptotanshinone, tanshinone I and tanshinone II A from Salvia Miltiorrhiza Bunge. 0.233 mg g(-1), 0.695 mg g(-1) and 0.682 mg g(-1) of each, respectively, were extracted using [OMIM][Cl], and preconcentrated in a [OMIM][PF(6)] phase at respective concentrations of 148.1, 507.1 and 486.1 MUg mL(-1). The method exhibited potential applicability with other medicinal products. PMID- 21645762 TI - Fabrication of column chip made of PMMA for MUFIA. AB - We proposed a low cost fabrication procedure of a poly(methylmethacrylate) (PMMA) column chip. 3D microchannel structure consisting of four columns in a chip for a mother die was fabricated using dry film photoresist and photolithography technique. Electroforming was applied to the mother die in order to obtain a Ni mold, then, the pattern was transferred to PMMA by hot press. The column had a dam structure to keep enzyme-immobilized microbeads with volume of 640 nL. The column chip was applied for a micro flow injection analysis (MUFIA) system. For a demonstration, we measured lactose using two columns in series. One column was set on upper stream and filled with chitosan microbeads immobilized with beta galactosidase, the other was on downstream and filled with the beads immobilized with glucose oxidase. The lactose detection was accomplished less than 90s after the sample injection. The biosensing system also showed a high performance for lactose detection in wide range of 1 MUM to 1mM. These results show that the column chip and our microfluidic biosensing system have the potential to assist minuaturization with small sample volume and short determination time for a sequential analysis. PMID- 21645763 TI - A simple and sensitive fluorescent sensing platform for Hg2+ ions assay based on G-quenching. AB - In this work, a novel fluorescence biosensor was demonstrated for detection of Hg(2+) ions with relatively high selectivity and sensitivity. The sensing scheme was based on G-quenching induced by Hg(2+) ions. In the presence of Hg(2+) ions, the single-stranded signal probe which has carboxylfluorescein (FAM) and guanine segment at its 5' and 3' ends, respectively, folded into duplex-like structure via the Hg(2+)-mediated coordination of T-Hg(2+)-T base pairs. It brought guannine segment close to the dye and caused a remarkable decrease of fluorescence signal. The sensor showed a sensitive response to Hg(2+) ions in a concentration range from 0.5 to 10 MUM, and a detection limit of 0.5 nM was given. This homogeneous system required only a single-labeled oligonucleotide, operated by concise procedures, and possessed comparable sensitivity as previous approaches. Furthermore, the sensor exhibits a great perspective for future practical applications. PMID- 21645764 TI - Metabolomic analysis of normal (C57BL/6J, 129S1/SvImJ) mice by gas chromatography mass spectrometry: detection of strain and gender differences. AB - Previous studies have shown that the C57 and 129 strains of mice display marked differences in behavioural performance, neuroanatomy, neurochemistry and synaptic plasticity. However, few metabolomic studies of their biofluids have been performed. As part of a series of metabolic phenotyping, the effects of gender and strain upon serum metabolite composition and variation are examined in this study using gas chromatography-mass spectrometry (GC-MS) in normal C57BL/6J and 129S1/SvImJ strains of mice. The 129S1/SvImJ strain is phenotypically distinct from the C57BL/6J strain and characteristic metabotypes are produced for both male and female mice of each strain. These data demonstrate that the C57BL/6J and 129S1/SvImJ strains of mice show a wide range of metabolic differences across glycine, serine and threonine metabolism; valine, leucine and isoleucine biosynthesis; and tricarboxylic acid cycle pathways. Remarkably, the concentration of glyceric acid in the 129S1/SvImJ strain is significantly increased compared to the C57BL/6J mouse strain, reflecting important considerations for studies that use the 129S1/SvImJ mouse as the human d glycericaciduria model. We infer that a deficiency of d-glycerate kinase would explain such a glyceric acid accumulation in the 129S1/SvImJ strain. More importantly, this differential metabolite level data provide insight into specific metabolic pathways and lay the groundwork for integrated studies of the mouse models. PMID- 21645765 TI - Label free DNA detection based on gold nanoparticles quenching fluorescence of Rhodamine B. AB - A novel and sensitive label free DNA detection method using gold nanoparticles (GNPs) and Rhodamine B (RB) has been developed. The assay is based on the following two properties. One is the different adsorption properties of single stranded and double-stranded DNA on GNPs in colloidal solution. The other is the different quenching ability of aggregated GNPs and dispersed GNPs on RB. Un aggregated GNPs could effectively quench the fluorescence of RB. However, the quenching ability greatly decreases after GNPs aggregated. The hybridization of probe DNA and target DNA is monitored by the fluorescence detection after the RB is added to the solution. Under the optimal experimental conditions, the detection limit of this assay is 2.9*10(-13) mol L(-1). PMID- 21645766 TI - Electrochemical aptasensor for detection of copper based on a reagentless signal on architecture and amplification by gold nanoparticles. AB - A highly sensitive and specific electrochemical aptasensor for Cu(2+) detection based on gold nanoparticles (AuNPs) is presented. In this work, AuNPs offered a big surface area to immobilize a large number of aptamers and excellent electrochemical signal transduction. Its high sensitivity, low detection limit, and wide detection range are the main advantages over our former copper aptasensor. The peak current increased proportionally to the Cu(2+) concentration over the range from 0.1 nM to 10 MUM with a detection limit of 0.1 pM. The presence of other divalent metal ions did not affect the detection of Cu(2+), which indicates a high specificity of Cu(2+) detection could be detected. Rapidity, simplicity, and excellent selectivity make it suitable for practical use in determination of Cu(2+) from lake samples. PMID- 21645767 TI - A high selective immunochromatographic assay for rapid detection of aflatoxin B1. AB - To solve the problem of low selectivity of current immunochromatographic assay (ICA) for aflatoxin B(1) (AFB(1)) alone detection, a novel selective ICA was developed here. With very high selectivity, a new AFB(1) monoclonal antibody (MAb) 3G1 was prepared by immunizing Balb/c mice with aflatoxin B(2a)-BSA (AFB(2a)-BSA) rather than AFB(1)-BSA used in other reports and 3G1 possessed the highest selectivity than those used in published ICAs. The ICA with visual detection limit (VDL) of 1 ng mL(-1) showed no cross-reactivity with other aflatoxins. Comparing with previous reports, the ICA here provided the most powerful guarantee for avoiding false positive results leaded by coexistence of other aflatoxins in samples. For validation, naturally contaminated samples including peanut, puer-tea, vegetable oil and feedstuff were respectively assayed by ICA and a standard high performance liquid chromatography (HPLC), and good agreement of results was obtained between two methods. Therefore, the developed ICA could well meet the selective detection of AFB(1) in agro-products. PMID- 21645768 TI - High-throughput dynamic microwave-assisted extraction on-line coupled with solid phase extraction for analysis of nicotine in mushroom. AB - A simple and low-cost high-throughput dynamic microwave-assisted extraction (HTDMAE) device was firstly assembled and validated by the extraction of nicotine in mushroom samples. In this device, a household microwave oven was applied to provide the microwave energy; a vacuum pump was used to deliver the solvent. Compared with traditional dynamic microwave-assisted extraction method, the sample throughput and microwave energy utilization were improved by the HTDMAE, up to 20 samples could be treated simultaneously in 9 min. Taking extraction of nicotine in mushroom sample as an example, a method was established with extraction, separation and enrichment of nicotine in a single step by the device on-line coupled with solid-phase extraction (SPE). Nicotine was first extracted from the mushroom samples with water under the action of microwave energy, and then directly introduced into the SPE column which was packed with cation exchange resins. Subsequently, the nicotine trapped on the resins was eluted with methanol-ammonia (95:5, v/v) and determined by high-performance liquid chromatography. The limit of detection of nicotine obtained is 5.6 MUg kg(-1) in fresh mushroom sample. The recovery of nicotine in mushroom samples is in the range of 87.4-104.0%. The proposed method which significantly reduced the overall analysis time and increased sample throughput should be favored for routine analyse of complex solid sample. PMID- 21645769 TI - Diffuse reflectance determination of Malachite Green using polyurethane foam as solid support and sodium dodecylsulfate as counter ion. AB - This paper reports the use of polyurethane foam (PUF) as solid support for diffuse reflectance spectrophotometric determination of Malachite Green (MG), a well known cationic dye used as biocide in the aquaculture industry, using sodium dodecylsulfate (SDS) as a counter ion. The method was based on the formation of an ionic-pair between the dye and the anionic surfactant SDS, which was sorbed onto PUF surface, where the diffuse reflectance was measured at 635 nm. Several parameters that could affect the performance of the system were evaluated. As expected, the SDS concentration presented strong influence on the analytical signal because the PUF was able to retain only the ionic-pair. The pH influenced the analytical signal, which was more intense in the acidic/neutral range, while the ionic strength only influenced the kinetic of the MG (as MG-SDS ionic-pair) sorption. The methodology was employed in the determination of MG in river waters and a recovery test was performed to test the accuracy of the procedure. Recovery percentages between 98.7 and 107% were observed when 60 or 80 MUg L(-1) of MG were added to the samples. PMID- 21645770 TI - Electrochemical response of ascorbic and uric acids at organoclay film modified glassy carbon electrodes and sensing applications. AB - A naturally occurring Cameroonian smectite clay has been grafted with trimethylpropylammonium groups and the resulting organoclay deposited as thin film onto a glassy carbon electrode (GCE) surface. It was then exploited as a suitable matrix for the accumulation and the electrochemical detection of ascorbic acid (AA) and uric acid (UA). Cyclic voltammetry revealed an increase in oxidation peak responses along with a negative shift of the corresponding anodic peak potential for both AA and UA species when using the organoclay coated GCE in comparison with the bare electrode. The electroanalytical response was improved by coating the electrode surface with a first layer of sublimed ferrocene (FC(s)), and then overcoating with the organoclay film to avoid the mediator leaching. The resulting bilayer film exhibited good characteristics such as extended linear range and high sensitivities for AA and UA, in cyclic voltammetry and amperometry. Interestingly, the redox mediator FC(s) was likely to lower overpotentials for AA oxidation (but not for UA), making possible the selective detection of these species in a mixture. The developed method could be used for the determination of AA in a pharmaceutical preparation and for UA in urine. PMID- 21645771 TI - Sulfur-nanoparticle-based method for separation and preconcentration of some heavy metals in marine samples prior to flame atomic absorption spectrometry determination. AB - The application of sulfur-nanoparticle-loaded alumina as an efficient adsorbent for the solid-phase extraction (SPE) and determination of trace amounts of Cd, Cu, Zn, and Pb ions was investigated in marine samples using flame atomic absorption spectrometry (FAAS). The nanometer-sized sulfur particles were synthesized in situ, physically loaded onto alumina microparticles, and the parameters influencing the preconcentration of the analytes, such as the pH, solution flow rate and volume, eluent solution, and interfering ions, were examined. The results showed that the optimal conditions for quantitative recovery of the metal ions by adsorption and elution on the sulfur nanoparticles (SNPs) was achieved by employing a flow rate of 15 mL min(-1), a pH of 8.5 for the sample solutions, and an eluent composed of 3.0 mol L(-1) HNO(3) in methanol. The detection limits of this method for Cd, Zn, Cu, and Pb ions were 0.30, 0.21, 0.24, and 0.63 MUg L(-1) (n=10), respectively. Application of the proposed method to the analysis of fish certified reference material (DORM-3) produced results that were in good agreement with the certified values. The proposed method was also successfully applied to the determination of analytes in marine samples, including seawater, fish, and oysters. PMID- 21645772 TI - Electrochemical analysis of HIV-1 reverse transcriptase serum level: exploiting protein binding to a functionalized nanostructured surface. AB - This manuscript describes an electrochemical approach to the detection of the reverse transcriptase of the human immunodeficiency virus type-1 (HIV-1 RT) in serum exploiting an organometallic peptide conjugate that is chemically linked to a nanostructured gold surface. The assay format is based on the formation of a thin film of a ferrocene-labeled lipoic acid (Fc-LA) onto a gold nanoparticles functionalized screen-printed carbon electrode (GNPs-SPCE). Time-of-Flight secondary ion mass spectrometry (TOF-SIMS) and X-ray photoelectron spectroscopy were employed to confirm the binding of the Fc-LA to the electrode surface via formation of a gold-thiol bond. The RT biosensor was developed by covalent attachment of the peptide VEAIIRILQQLLFIH to the carboxylic acid group of Fc-LA. Square wave voltammetry offered a two-dimensional measurement of RT based on the anodic shift and reduction of current density of the Fc redox signal upon binding of RT to its specific peptide. This allowed a linear quantification of the target RT in the range of 1-500 pg mL(-1) equivalent to 0.9-427 fM, with a detection limit of 0.8 pg mL(-1) (0.7 fM) with a short response time. PMID- 21645773 TI - Sensitive and selective off-on rhodamine hydrazide fluorescent chemosensor for hypochlorous acid detection and bioimaging. AB - A rhodamine 6G hydrazide fluorescent chemosensor was prepared for the rapid HOCl detection in aqueous media. The system makes good use of the irreversible HOCl mediated selective oxidation reaction to generate fluorescent response proportional to the amount of HOCl in neutral buffer. This probe exhibits great photostability, high sensitivity, and good selectivity for HOCl over other reactive species and most of the common metal ions. Furthermore, the probe is cell membrane permeable, and its applicability has been successfully demonstrated for fluorescence imaging of both exogenous and endogenous HOCl within living cells. Cytotoxicity assays prove that this probe is almost nontoxic to the cultured cell lines under the experimental conditions. PMID- 21645774 TI - Ionic liquid sensitized fluorescence determination of four isoquinoline alkaloids. AB - The fluorescence spectra of berberine, palmatine, jatrorrhizine, and coptisine in ionic liquids were studied and found to increase significantly in ionic liquids, with [C(8)MIM][PF(6)] having the greatest increase. Further studies showed that these drugs could be extracted from an aqueous solution by [C(8)MIM][PF(6)] using the temperature-assisted ionic liquid dispersive liquid phase microextraction method. The enrichment factors were 81.8-82.3, and the extraction recovery was 98.5%, 98.1%, 98.3%, and 98.8% for berberine, palmatine, jatrorrhizine, and coptisine, respectively. Based on the [C(8)MIM][PF(6)] preconcentration, separation, and sensitized fluorescence for these drugs, a new selective and sensitive method for the determination of concentration of these four drugs in aqueous samples was presented. At optimum conditions, the linear relationship was obtained in the ranges of 0.8-130 ng mL(-1), 0.9-160 ng mL(-1), 0.7-140 ng mL( 1), and 0.6-110 ng mL(-1), respectively. The proposed method was successfully applied for the determination of the drugs in pharmaceutical preparations, urine, and plasma samples. PMID- 21645775 TI - A simple and demountable capillary microflow nebulizer with a tapered tip for inductively coupled plasma mass spectrometry. AB - A simple and demountable capillary microflow nebulizer (d-CMN) was developed for inductively coupled plasma mass spectrometry (ICP-MS). It consisted of a nebulizer body, a fused-silica capillary with a tapered tip and a polytetrafluoroethylene (PTFE) adapter. The gas orifice i.d., the solution capillary tip i.d. and its wall thickness were 200, 30, and 5 MUm, respectively. The sensitivities, detection limits, precisions and the long-term stability with the d-CMN were evaluated. The experimental results indicated that its performances at low uptake rates were similar or better than those obtained with the conventional concentric nebulizer at 820 MUL/min and the micromist nebulizer at 200 MUL/min. The demountable construction of the d-CMN permitted that the blocked or broken solution capillary could be conveniently renewed. The low self aspiration rate (4.77 MUL/min) and the analytical characteristics comparable to commercial microflow nebulizers made the d-CMN a good choice for coupling capillary electrophoresis and microbore high-performance liquid chromatograph to ICP-MS. The proposed d-CMN was successfully applied in the iodine speciation by coupling microchip capillary electrophoresis (MCE) to ICP-MS. The absolute detection limits for iodide and iodate of 0.20 and 0.29 fg were achieved with satisfactory resolution. PMID- 21645776 TI - Alert-type biological dosimeter based on enzyme logic system. AB - A cooperative effect of two biomarkers, alpha-amylase and lactate dehydrogenase, was used to analyze radiation-caused tissue damage in vitro in model solutions of human serum. The analytical system was based on the recently emerged biocomputing concept applying biocatalytic cascades for logic processing of biochemical input signals. The studied system resembled a Boolean NAND logic gate in which the change of the optical output signal from a high level (logic value 1) to a low level (logic value 0) confirmed the presence of both biomarkers at pathological concentrations (1,1 input signals), thus yielding the conclusion about radiation tissue damage. The system operates in a digital YES/NO format as an alert-type biosensor with a built-in Boolean logic. PMID- 21645777 TI - Sequential injection Lab-at-valve (SI-LAV) segmented flow system for kinetic study of an enzyme. AB - A sequential injection-Lab-at-valve (SI-LAV) segmented flow system for kinetic study of an enzyme was developed. Air segments were introduced for separation of enzyme and substrate zones and separation of the stacked zones from the carrier solution which ensure the measurement of the initial rate and minimize the dilution/dispersion effect. The open- ended mixing chamber makes it possible to use air segments in the flow system without the need for additional air segment discarding steps. The enzyme horseradish peroxidase (HRP) kinetic parameters based on initial rate was used as a model study. The operation of the system is virtually the same as that of the conventional batch-wise process. The kinetic parameters (i.e. K(m) and V(max)) of HRP obtained using the proposed system agree well with those obtained using the batch-wise process as well. The proposed system offers additional benefits of volume down scaling, improved rapidity and automatic features that does not require a skillful operator. PMID- 21645778 TI - 4-Carbomethoxybenzaldehyde as a highly sensitive pre-column fluorescence derivatization reagent for 9,10-phenanthrenequinone. AB - 9,10-Phenanthrenequinone (PQ) is harmful environmental pollutant that is detected in airborne particulates. The measurement of PQ in the air should be necessary to evaluate the potential adverse effects of PQ on human health. We have recently developed a determination method for PQ based on the fluorescence derivatization of PQ using benzaldehyde and ammonium acetate as a reagent. In this study, in order to obtain more sensitive and selective fluorescence derivatization reaction, we measured the fluorescence of the reaction mixture of PQ with 21 kinds of aromatic aldehydes in the presence of ammonium acetate. Among the tested aldehydes, 4-carbomethoxybenzaldehyde was found to be the best reagent in regard to fluorescence intensity and emission wavelength maximum. Based on the fluorescence derivatization with 4-carbomethoxybenzaldehyde, a highly sensitive chromatographic method was developed for the determination of PQ with the detection limit (S/N=3) of 1.2 fmol/injection. PMID- 21645779 TI - Determination of dissociation constants of aristolochic acid I and II by capillary electrophoresis with carboxymethyl chitosan-coated capillary. AB - Aristolochic acid-I and aristolochic acid-II have been proved to be the main bioactive and toxic component in Aristolochia plants. As a result, the determination of their dissociation constants, which are important property parameters for weak acids, is highly desired for related pharmacological and toxicological studies. In this work, the dissociation constant values of aristolochic acid-I and aristolochic acid-II were determined by capillary electrophoresis using carboxymethyl chitosan-coated capillary, based on their electrophoretic mobilities by using nonlinear regression as well as linear regression, showing that the two models give comparable results. The data were also compared with those obtained by capillary electrophoresis with polybrene coated capillary, and no conspicuous difference was observed. The correlation coefficients were all higher than 0.998 for both linear and nonlinear regression model. The pKa values were found to be 3.3+/-0.1 for aristolochic acid-I and 3.2+/-0.1 for aristolochic acid-II. PMID- 21645780 TI - Label-free detection of HIV-2 antibodies in serum with an ultra-high frequency acoustic wave sensor. AB - Herein is described a label-free immunosensor dedicated to the detection of HIV 2. The biosensor platform is constructed as a mixed self-assembled monolayer coated quartz wafer onto which HIV-2 immunodominant epitopes are immobilized. The biosensing properties, in terms of specific vs. non-specific antigen-antibody interactions, are evaluated with the electromagnetic piezoelectric acoustic sensor (EMPAS) using equimolar serum solutions of HIV-2 or HIV-1 monoclonal antibodies, respectively. This immunosensor constitutes the first real-world application of the EMPAS technology in the bioanalytical field. PMID- 21645781 TI - Fishing potential antitumor agents from natural plant extracts pool by dialysis and G-quadruplex recognition. AB - Screening G-quadruplex ligands from natural plants is important because the ligands may be potential antitumor drugs. A new screening strategy is proposed based on the combination of dialysis and G-quadruplex recognition technique which could separates G-quadruplex ligand from natural extracts and elucidate the structure of this ligand. This result offers a novel approach to obtain active antitumor compounds. PMID- 21645782 TI - A minimal core based fluorophore for selective detection of Zn(II) ions in aqueous solution and living cells. AB - A minimal core based fluorophore was introduced as a selectively fluorescent "turn on" sensor for Zn(2+) ions in aqueous solution. Addition of Zn(2+) ions to the fluorophore generates a significant emission through a 1:1 ligand-to-metal complex. The fluorescence titration experiment of the minimal core based fluorophore with various metal ions shows that the pyromellitic diimide derivative also has the advantage of a high selectivity to Zn(2+) ions over other metals such as Ni(2+), or Co(2+), Cu(2+), Fe(3+), Fe(2+). More than 8 fold increase in the intensity of fluorescence was observed for the Zn(2+)-bound fluorophore compared to Zn-free fluorophore. Due to its small molecular size, the fluorophore was cell-permeable and successfully applied to the detection of Zn(2+) in living cells. With its relatively high sensitivity to Zn(2+) in living cells, the synthesized new fluorophore will be very useful in the studies on various biological functions of Zn(2+). PMID- 21645783 TI - Multicolor saccharide-sensing chips based on boronic acid-containing thin films showing stepwise release and binding of dyes. AB - A novel saccharide sensor that shows a distinct color change resembling a "traffic signal" was developed. By copolymerizing a boronic acid monomer and an amine monomer on a glass plate, a boronic acid-containing thin film was obtained. After adsorbing anionic blue and yellow dyes, the thin film was immersed in aqueous saccharide solutions containing a cationic red dye. As saccharide concentration increased, the thin film changed its color from green to red via yellow. Origin of the distinct color change was attributed to a stepwise release and binding of dyes. PMID- 21645785 TI - Physical activity or academic achievement? Both! PMID- 21645786 TI - Foreword: the tug of war between vaginal and cesarean births. PMID- 21645787 TI - Delivery after previous cesarean. Preface. PMID- 21645788 TI - Recent trends and patterns in cesarean and vaginal birth after cesarean (VBAC) deliveries in the United States. AB - Cesarean delivery is the most common major surgical procedure for women in the United States, with 1.4 million surgeries annually. In 2008, nearly one-third (32.3%) of US births were by cesarean delivery. Cesarean delivery rates have increased rapidly in the United States in recent years because of an increasing primary cesarean delivery rate and a declining vaginal birth after cesarean (VBAC) rate. In 2007, the VBAC rate was 8.3% in a 22-state reporting area. The US VBAC rate was lowest among 14 industrialized countries; 3 countries had VBAC rates greater than 50%. PMID- 21645789 TI - Nonclinical factors affecting women's access to trial of labor after cesarean delivery. AB - The use of trial of labor after cesarean (TOLAC) has declined in the last decade, and the clinical risks of TOLAC remain low. Nonclinical factors continue to affect women's access to TOLAC. This article considers 5 categories of factors that seem to be influencing rates of TOLAC and vaginal birth after cesarean: opinion leaders and professional guidelines, hospital facilities and cesarean availability, reimbursement for providing TOLAC, medical liability, and patient level factors. An evidence base and strategies to provide guidance to create a safe environment for vaginal birth after cesarean are needed. Obstetric information systems are critical to this effort. PMID- 21645790 TI - VBAC: a medicolegal perspective. AB - History has always been a series of pendulum swings, and there is perhaps no better example in obstetrics than that of vaginal birth after cesarean. Vaginal birth after cesarean (VBAC) rates rose steadily in the early 1990s. However, VBAC rates have declined dramatically over recent years, while the cesarean delivery rate has continued to rise unabated. Many physicians and hospitals are no longer offering trial of labor after cesarean, largely because of medicolegal concerns. This article explores the medical and legal risks of trial of labor after cesarean. PMID- 21645791 TI - An ethical framework for the informed consent process for trial of labor after cesarean delivery. AB - In 2010, a National Institutes of Health Consensus Panel and the American College of Obstetricians and Gynecologists issued updated statements on trial of labor after cesarean delivery (TOLAC). This article presents an ethical framework for the informed consent process for TOLAC. Three conclusions are reached. For women with one previous low transverse incision, TOLAC and elective repeat cesarean delivery should be offered. Obstetricians should recommend against TOLAC when a pregnant woman has had a previous classical incision. TOLAC after two previous low transverse incisions may be offered provided that the informed consent process presents the uncertainties of the evidence. PMID- 21645792 TI - Delivery after prior cesarean: success rate and factors. AB - Cesarean delivery rates in the United States have reached an all-time high. The current rate of 31% is 6 times higher than the 1970s rate. Many factors including physician preference and hospital accessibility account for this trend. A decreased vaginal birth after cesarean (VBAC) rate and an increased repeat cesarean rate have important consequences for women in future pregnancies. Because of these considerations, VBAC has been an important issue within the obstetric community for over 3 decades. Identifying the best candidates for VBAC using factors available to the obstetrician can increase the VBAC success rate while minimizing maternal morbidity. PMID- 21645793 TI - Can a vaginal birth after cesarean delivery be a normal labor and birth? Lessons from midwifery applied to trial of labor after a previous cesarean delivery. AB - Women who undergo a trial of labor after a previous cesarean delivery (TOLAC) have special needs prenatally and during the intrapartum period. Counseling about the choice of TOLAC versus an elective repeat cesarean delivery involves complex statistical concepts. Prenatal counseling that is patient centered, individualized, and presented in a way that addresses the health literacy and health numeracy of the recipient encompasses best practices that support patient decision making. Evidence-based practices during labor that support vaginal birth and increase patient satisfaction are of special value for this population. PMID- 21645794 TI - The influence of intrapartum factors on risk of uterine rupture and successful vaginal birth after cesarean delivery. AB - Cesarean delivery is common and increasing over time. A prior cesarean birth increases the risk of both elective and emergency cesarean births and uterine rupture in a subsequent pregnancy. A range of factors, including labor characteristics, may influence the risk of these outcomes in the next pregnancy. Intrapartum factors associated with successful vaginal birth and lower risk of uterine rupture include the spontaneous onset of labor and advanced cervical dilatation. In contrast, need for induction and augmentation of labor are both factors associated with an increased likelihood of unsuccessful vaginal birth and risk of uterine rupture. PMID- 21645795 TI - Uterine rupture during a trial of labor after previous cesarean delivery. AB - Uterine rupture, which involves complete separation of the uterine wall, occurs in about 1% of those attempting vaginal birth after cesarean. Because uterine rupture is one of the most significant complications of a trial of labor (TOL) after previous cesarean, identifying those at increased risk of uterine rupture is paramount to the safety of a TOL after previous cesarean birth. It seems that both antepartum demographic characteristics and intrapartum factors modify the risk of uterine rupture. The ability to reliably predict an individual's a priori risk for intrapartum uterine rupture remains a major area of investigation. PMID- 21645796 TI - Multiple repeat cesareans and the threat of placenta accreta: incidence, diagnosis, management. AB - Placenta accreta is a significant source of obstetric morbidity and mortality. Its incidence is increasing as a direct consequence of the increasing cesarean section rate, which reflects increased rates of maternal obesity, increased numbers of multiple gestations secondary to assisted reproductive technology, physician concern about litigation for adverse obstetric outcome, and a decline in the use of operative vaginal delivery for both cephalic and breech presentations. Optimum management for most cases requires elective cesarean hysterectomy, ideally performed at about 34 weeks' gestation. A multidisciplinary approach produces the best outcomes. PMID- 21645797 TI - Delivery after prior cesarean: maternal morbidity and mortality. AB - Nearly 1 in 3 pregnant women in the United States undergo cesarean. This trend is contrary to the national goal of decreasing cesarean delivery in low-risk women. The decline in vaginal birth after cesarean (VBAC) contributes to the continual increase in cesarean deliveries. Prior cesarean delivery is the most common indication for cesarean and accounts for more than one-third of all cesareans. The appropriate use and safety of cesarean and VBAC are of concern not only at the individual patient and clinician level but they also have far-reaching public health and policy implications at the national level. PMID- 21645798 TI - Fetal and neonatal morbidity and mortality following delivery after previous cesarean. AB - This article examines data from a recent systematic evidence review on term deliveries conducted for the National Institutes of Health Consensus Conference sponsored by the Agency for Healthcare Research and Quality on vaginal birth after caesarean, from a meta-analysis of associated perinatal outcomes, and subsequent publications that meet stringent quality review standards. We present a summary of fetal and neonatal outcomes emphasizing information that clinicians and patients need to make decisions regarding mode of delivery after prior cesarean and look for areas where future studies may provide important insights. PMID- 21645800 TI - Geriatrics: why do we do it? PMID- 21645799 TI - Cesarean versus vaginal delivery: long-term infant outcomes and the hygiene hypothesis. AB - Concurrent with the trend of increasing cesarean delivery numbers, there has been an epidemic of both autoimmune diseases and allergic diseases. Several theories have emerged suggesting that environmental influences are contributing to this phenomenon, most notably, the hygiene hypothesis. This article provides background about the human microbiota and its relationship to the developing immune system as well as the relationship of mode of delivery on the colonization of the infant intestine, development of the immune system, and subsequent childhood allergies, asthma, and autoimmune diseases. PMID- 21645801 TI - Anticoagulant polypharmacy. PMID- 21645802 TI - Incentive payments: extra payments for Medicare services. PMID- 21645803 TI - Falls: why they matter and what you can do. PMID- 21645804 TI - Recognizing and referring suspected elder mistreatment. PMID- 21645805 TI - Adiposity and adiponectin in dogs: investigation of causes of discrepant results between two studies. AB - Although one study showed lower adiponectin concentrations in obese dogs, other recent studies indicate that adiponectin might not be decreased in obese dogs, raising the possibility that the physiology of adiponectin is different in dogs than in humans. The aim of this study was to investigate possible causes of the discrepancy between the two largest studies to date that assessed the association between adiposity and adiponectin concentration in dogs, including the validity of the assay, laboratory error, and the effects of breed, sex, and neuter status on the relationship between adiposity and adiponectin concentrations. Adiponectin concentrations measured with a previously validated adiponectin ELISA were compared with those estimated by Western blotting analysis of reduced and denatured plasma samples. The possibility of laboratory error and the effect of EDTA anticoagulant and aprotinin were tested. Adiponectin concentration was measured by ELISA in 20 lean dogs (10 male and 10 female, 5 neutered in each sex). There was close correlation between adiponectin concentrations measured by ELISA and those estimated by Western blotting analysis (r = 0.90; P < 0.001). There was no substantial effect of EDTA, aprotinin, or laboratory error on the results. There was confounding by neuter status of the relationship between adiposity and adiponectin concentrations, but adiponectin concentrations were not significantly lower in male than in female lean dogs (females, 36 mg/L; males, 26 mg/L; P > 0.20) and were not significantly lower in intact than in neutered lean male dogs (intact, 28 mg/L; neutered, 23 mg/L; P = 0.49). We conclude that the adiponectin ELISA previously validated for use in dogs appears to be suitable for determination of canine adiponectin concentrations and that testosterone does not appear to have a strong effect on plasma adiponectin concentrations in dogs. Obesity might decrease adiponectin concentrations in intact but not in neutered dogs. PMID- 21645806 TI - The GLP-1 mimetic exenatide potentiates insulin secretion in healthy cats. AB - The glucagon-like peptide-1 mimetic exenatide has a glucose-dependent insulinotropic effect, and it is effective in controlling blood glucose (BG) with minimal side effects in people with type 2 diabetes. Exenatide also delays gastric emptying, increases satiety, and improves beta-cell function. We studied the effect of exenatide on insulin secretion during euglycemia and hyperglycemia in cats. Nine young, healthy, neutered, purpose-bred cats were used in a randomized, cross-over design. BG concentrations during an oral glucose tolerance test were determined in these cats previously. Two isoglycemic glucose clamps (mimicking the BG concentration during the oral glucose tolerance test) were performed in each cat on separate days, one without prior treatment (IGC) and the second with exenatide (1 MUg/kg) injected subcutaneously 2 h before (ExIGC). BG, insulin, and exenatide concentrations were measured, and glucose infusion rates were recorded and compared in paired tests between the two experiments. After exenatide injection, insulin serum concentrations increased significantly (2.4 fold; range 1.0- to 9.2-fold; P = 0.004) within 15 min. This was followed by a mild decrease in BG concentration and a return of insulin concentration to baseline despite a continuous increase in serum exenatide concentrations. Insulin area under the curve (AUC) during ExIGC was significantly higher than insulin AUC during IGC (AUC ratio, 2.0 +/- 0.4; P = 0.03). Total glucose infused was not significantly different between IGC and ExIGC. Exenatide was detectable in plasma at 15 min after injection. The mean exenatide concentration peaked at 45 min and then returned to baseline by 75 min. Exenatide was still detectable in the serum of three of five cats 8 h after injection. No adverse reactions to exenatide were observed. In conclusion, exenatide affects insulin secretion in cats in a glucose dependent manner, similar to its effect in other species. Although this effect was not accompanied by a greater ability to dispose of an intravenous glucose infusion, other potentially beneficial effects of exenatide on pancreatic beta cells, mainly increasing their proliferation and survival, should be investigated in cats. PMID- 21645807 TI - Plasma ghrelin concentration is decreased by short chain fatty acids in wethers. AB - To examine the effects of short chain fatty acids (SCFAs) on plasma ghrelin concentration, 4 wethers were injected intravenously with SCFA solutions [acetate (ACE), propionate (PRO), and butyrate (BUT) (0.8 mmol/kg BW)] and saline. The experiment was conducted after a 4 * 4 Latin square design. Each solution was injected into the jugular vein catheter with blood samples taken at -10, 0, 5, 10, 15, 20, 25, 30, 40, 50, and 60 min relative to the injection time also from this catheter. Plasma ghrelin concentrations decreased after injection with ACE, PRO, and BUT. Although plasma glucose concentrations increased after injection with PRO and BUT (P < 0.05), the increment areas were greater with BUT than with PRO. Plasma insulin concentrations increased after injection with PRO and BUT (P < 0.05). The decrement areas in plasma ghrelin concentrations were equal in ACE, PRO, and BUT. These data suggest that SCFAs inhibit ghrelin secretion in wethers and not through increased circulating glucose and insulin as previously proposed. PMID- 21645808 TI - Salting of dry-cured meat - A potential cause of contamination with the ochratoxin A-producing species Penicillium nordicum. AB - Penicillium nordicum is a known contaminant of protein-rich foods and is primarily found on dry-cured meat products. It is an important producer of the mycotoxin ochratoxin A, which has nephrotoxic and cancerogenic activities. Recently a high number of P. nordicum strains was isolated from different dry cured meat products from one of the Slovenian meat-processing plants. Since we have isolated P. nordicum in high counts also from Artic habitats, such as sea water and sea ice and due to its ability to grow well at low temperatures and at increased salinity, sea salt was suspected as the possible source of P. nordicum. In the present study contamination of meat products, air in the meat-processing plant and sea salt used for salting were analysed. When 50 g of salt sample from a sealed package was dissolved in sterile water and filtered, 12 colonies of P. nordicum were obtained on solid medium incubated at 15 degrees C, while a salt sample from an open vessel in the meat-processing area developed high, uncountable number of colonies. Amplified fragment length polymorphism analyses of P. nordicum isolates from different sources showed that contamination of meat products via salt was possible. Three selected isolates examined for extrolites all produced ochratoxin A. As contamination of dry-cured meat products with P. nordicum represents a potential health risk for consumers and workers in the meat processing plants, salt should be taken into account as a potential cause of such contaminations. PMID- 21645809 TI - Evaluation of ISO 10272:2006 standard versus alternative enrichment and plating combinations for enumeration and detection of Campylobacter in chicken meat. AB - In the present study, we evaluate the recommended ISO 10272:2006 versus alternative procedures for Campylobacter enumeration and enrichment in naturally contaminated chicken meat samples (n = 49). Three enrichment media were evaluated; Bolton broth, Preston broth and CampyFood broth((r)) (bioMerieux SA, Marcy l'Etoile, France). In addition, three selective plating agars were compared; modified charcoal cefoperazone deoxycholate agar (mCCDA), CampyFood agar((r)) (CFA; bioMerieux SA) and Brilliance CampyCount agar((r)) (BCC; Oxoid, Basingstoke, England). Direct plating on CFA provided the highest number of Campylobacter positive samples (17/49); however this was not statistically different (P > 0.05) from numbers of positive samples recovered by direct plating on mCCDA (15/49) or BCC agars (14/49). Also, there was no significant difference between Campylobacter counts on the three compared media (P > 0.05). The coloured colonies of Campylobacter on CFA and BCC were easier to record and count than those on mCCDA. Enrichment of chicken meat samples in Bolton broth for 48 h and subsequent plating on CFA provided significantly higher (P < 0.05) Campylobacter detection compared to the other broth-agar combinations. Enrichment in Preston broth for 24 h followed by plating on mCCDA gave a higher number of positive samples (20/49) than 48 h enrichment in Bolton broth and plating on mCCDA (15/49). Enrichment in Bolton broth for 48 h followed by plating on CFA recovered 35% of samples below the limit for quantifications (<10 CFU/g, n = 34), as identified by direct plating on mCCDA. Compared to the current ISO method, some alternative combinations of enrichment and agar media could provide significantly better detection and enumeration of Campylobacter in chicken meat. PMID- 21645810 TI - Molecular characterization of Salmonella enterica serovar Saintpaul isolated from imported seafood, pepper, environmental and clinical samples. AB - A total of 39 Salmonella enterica serovar Saintpaul strains from imported seafood, pepper and from environmental and clinical samples were analyzed for the presence of virulence genes, antibiotic resistance, plasmid and plasmid replicon types. Pulsed-field gel electrophoresis (PFGE) fingerprinting using the XbaI restriction enzyme and plasmid profiling were performed to assess genetic diversity. None of the isolates showed resistance to ampicillin, chloramphenicol, gentamicin, kanamycin, streptomycin, sulfisoxazole, and tetracycline. Seventeen virulence genes were screened for by PCR. All strains were positive for 14 genes (spiA, sifA, invA, spaN, sopE, sipB, iroN, msgA, pagC, orgA, prgH, lpfC, sitC, and tolC) and negative for three genes (spvB, pefA, and cdtB). Twelve strains, including six from clinical samples and six from seafood, carried one or more plasmids. Large plasmids, sized greater than 50 kb were detected in one clinical and three food isolates. One plasmid was able to be typed as IncI1 by PCR-based replicon typing. There were 25 distinct PFGE-XbaI patterns, clustered to two groups. Cluster A, with 68.5% similarity mainly consists of clinical isolates, while Cluster C, with 67.6% similarity, mainly consisted of shrimp isolates from India. Our findings indicated the genetic diversity of S. Saintpaul in clinical samples, imported seafood, and the environment and that this serotype possesses several virulent genes and plasmids which can cause salmonellosis. PMID- 21645811 TI - Prevalence and impact of single-strain starter cultures of lactic acid bacteria on metabolite formation in sourdough. AB - Flavour of type II sourdoughs is influenced by the ingredients, processing conditions, and starter culture composition. It is, however, not fully clear to what extent different sourdough lactic acid bacteria (LAB) contribute to flavour. Therefore, two types of flour (rye and wheat) and different LAB starter culture strains were used to prepare sourdoughs, thereby leaving the yeast microbiota uncontrolled. All LAB starter culture strains tested were shown to be prevalent and to acidify the flour/water mixture to pH values between 3.1 and 3.9 after 24h of fermentation. Multiple aldehydes, alcohols, ketones, and carboxylic acids were produced by the sourdough-associated microbiota throughout the fermentation period. Based on the organoleptic evaluation of breads produced with these sourdoughs, five LAB strains were selected to perform prolonged wheat and rye fermentations as to their capacity to result in an acidic (Lactobacillus fermentum IMDO 130101, Lactobacillus plantarum IMDO 130201, and Lactobacillus crustorum LMG 23699), buttermilk-like (Lactobacillus amylovorus DCE 471), or fruity flavour (Lactobacillus sakei CG1). Upon prolonged fermentation, higher metabolite concentrations were produced. For instance, L. sakei CG1 produced the highest amounts of 3-methyl-1-butanol, which was further converted into 3 methylbutyl acetate. The latter compound resulted in a fruity banana flavour after 48h of fermentation, probably due to yeast interference. Rye fermentations resulted in sourdoughs richer in volatiles than wheat, including 3-methyl-1 butanol, 2-phenylethanol, and ethyl acetate. PMID- 21645812 TI - The determinacy of reproducibility assessments of qualitative microbial food borne pathogen methods detecting a few microbes per analytical portion. AB - A summary of an examination of studies of interlaboratory reproducibility of measurements for detecting the presence of microorganisms in food products is presented. In such studies multiple laboratories, 10 or more, compare the performances of reference and test methods at the limit of detection of the methods that is at spiking levels around 1 cfu per analytical portion. A laboratory's performance is expressed as the number of positive replicates detected per set of six. The data only imply the presence or absence of significant between-laboratory effects with the test method relative to the reference method. It is difficult to parse the observed variability into the contributions of the sample variability and between-laboratory effects. This is because at spiking levels close to 1 cfu per portion it cannot be assured that laboratories are examining portions with equivalent numbers of the target microbe. In this study published data are reformulated to the number of laboratories observing a given number of positive results per replicate set in order to reflect the inhomogeneity of the spike distribution in the replicate portions. A mean spiking level that is less uncertain than the reported 3-tube reference method MPN value is estimated from the pooled proportions of positives that the laboratories obtained with the reference method. The expected distribution of the spike was calculated from its mean value using the binomial equation. The numerical distributions of the laboratories among the 7 possible positive categories (0-6 positives per replicate set) were statistically indistinguishable from the expected binomial distribution thus corroborating this approach. Probable instances of interlaboratory performance differences were detected by further statistical analysis. This analytical approach, as well as transparently reflecting the sampling variability, also suggested ways of improving and simplifying such studies. PMID- 21645813 TI - PCR ITS-RFLP: A useful method for identifying filamentous fungi isolates on grapes. AB - Restriction digestion analysis of the ITS products was tested as an easy method to identify isolates of filamentous fungi on grapes. Endonucleases SduI, HinfI, MseI, HaeIII were used. Endonucleases BfmI, Cfr9I, Hpy188I, MaeII or PspGI were used as necessary to complete discrimination. The 43 species studied generated 42 different composite profiles. Only the species P. thomii and P. glabrum gave the same composite profile. 96.3% strains tested could be differentiated to the species level with only four enzymes. Hundred ninety nine strains of filamentous fungi were isolated from various vineyards in Burgundy and identified by this method. Penicillium (58.5%) was the genus the most frequently isolated and no strains of the genus Aspergillus was isolated. P. spinolusum was the most isolated species of Penicillium (22.70%). The species C. cladiosporioides, B. cinerea, E. nigrum, A. alternata, T. koningiopsis, P. diplodiella, C. herbarum, A. alternatum, T. cucumeris and F. oxysporum were also isolated. This technique is a rapid and reliable method appropriate for routine identification of filamentous fungi. This can be used to screen large numbers of isolates from various environments in a short time. This is the first exhaustive study of fungal diversity at species level in vineyard. PMID- 21645814 TI - Quantifying the individual effects of ethanol and temperature on the fitness advantage of Saccharomyces cerevisiae. AB - The presence of Saccharomyces cerevisiae in grape berries and fresh musts is usually very low. However, as fermentation progresses, the population levels of this species considerably increase. In this study, we use the concept of fitness advantage to measure how increasing ethanol concentrations (0-25%) and temperature values (4-46 degrees C) in wine fermentations affects competition between S. cerevisiae and several non-Saccharomyces yeasts (Hanseniaspora uvarum, Torulaspora delbrueckii, Candida zemplinina, Pichia fermentans and Kluyveromyces marxianus). We used a mathematical approach to model the hypothetical time needed for S. cerevisiae to impose itself on a mixed population of the non-Saccharomyces species described above. This approach also took into consideration the influence of environmental factors and the initial population levels of S. cerevisiae (0.1, 1.0 and 10.0%). Our results suggest that Saccharomyces niche construction via ethanol production does not provide a clear ecological advantage (at least not until the ethanol concentration exceeds 9%), whereas a temperature rise (above 15 degrees C) does give S. cerevisiae a considerable advantage. The initial frequency of S. cerevisiae considerably influences the time it needs to impose itself (until it reaches a final frequency of 99% in the mixed culture), the lowest time values being found at the highest initial frequency. In light of these results, the application of low temperatures in the wine industry could favor the growth and survival of non-Saccharomyces species for a longer period of time. PMID- 21645815 TI - Seafood quality analysis: Molecular identification of dominant microbiota after ice storage on several general growth media. AB - This study points out the limitations of several general growth media frequently used in seafood research by a systematic identification of the microorganisms on fish samples during ice storage unable to grow on those media. Aerobic psychrotrophic count (APC), replication on various general media and total cultivable microbial community denaturing gradient gel electrophoresis (DGGE) analysis revealed that many potential spoilage microorganisms were overlooked. Those microorganisms overlooked by using only one single growth medium were identified by partial 16S rRNA gene and gyrB gene sequencing. Members of the genera Shewanella, Vibrio, Aliivibrio, Photobacterium, Pseudoalteromonas and Psychrobacter, including Photobacterium phosphoreum, Shewanella baltica and Pseudomonas fluorescens are unable to grow on PCA. APC analysis also confirmed that on plate count agar (PCA) the enumeration of the microbiota was underestimated. Although Long and Hammer agar (LH) and marine agar (MA) obtained the best quantitative (APC analysis) and qualitative (replication and DGGE analyses) results for fish quality analysis, analysts have to keep in mind that some species were also unable to grow on those media, such as Pseudomonas fragi and Acinetobacter sp. PMID- 21645816 TI - Modeling the pulsed light inactivation of microorganisms naturally occurring on vegetable substrates. AB - Pulsed light (PL) is a fast non-thermal method for microbial inactivation. This research studied the kinetics of PL inactivation of microorganisms naturally occurring in some vegetables. Iceberg lettuce, white cabbage and Julienne-style cut carrots were subjected to increasing PL fluences up to 12J/cm(2) in order to study its effect on aerobic mesophilic bacteria determined by plate count. Also, sample temperature increase was determined by infrared thermometry. Survivors' curves were adjusted to several models. No shoulder but tail was observed. The Weibull model showed good fitting performance of data. Results for lettuce were: goodness-of-fit parameter RMSE=0.2289, fluence for the first decimal reduction delta=0.98+/-0.80J/cm(2) and concavity parameter p=0.33+/-0.08. Results for cabbage were: RMSE=0.0725, delta=0.81+/-0.23J/cm(2) and p=0.30+/-0.02; and for carrot: RMSE=0.1235, delta=0.39+/-0.24J/cm(2) and p=0.23+/-0.03. For lettuce, a log-linear and tail model was also suitable. Validation of the Weibull model produced determination coefficients of 0.88-0.96 and slopes of 0.78-0.99. Heating was too low to contribute to inactivation. A single low-energy pulse was enough to achieve one log reduction, with an ultrafast treatment time of 0.5ms. While PL efficacy was found to be limited to high residual counts, the achievable inactivation level may be considered useful for shelf-life extension. PMID- 21645817 TI - Monitoring the microbial community during solid-state acetic acid fermentation of Zhenjiang aromatic vinegar. AB - Zhenjiang aromatic vinegar is one of the most famous Chinese traditional vinegars. In this study, change of the microbial community during its fermentation process was investigated. DGGE results showed that microbial community was comparatively stable, and the diversity has a disciplinary series of changes during the fermentation process. It was suggested that domestication of microbes and unique cycle-inoculation style used in the fermentation of Zhenjiang aromatic vinegar were responsible for comparatively stable of the microbial community. Furthermore, two clone libraries were constructed. The results showed that bacteria presented in the fermentation belonged to genus Lactobacillus, Acetobacter, Gluconacetobacter, Staphylococcus, Enterobacter, Pseudomonas, Flavobacterium and Sinorhizobium, while the fungi were genus Saccharomyces. DGGE combined with clone library analysis was an effective and credible technique for analyzing the microbial community during the fermentation process of Zhenjiang aromatic vinegar. Real-time PCR results suggested that the biomass showed a "system microbes self-domestication" process in the first 5 days, then reached a higher level at the 7th day before gradually decreasing until the fermentation ended at the 20th day. This is the first report to study the changes of microbial community during fermentation process of Chinese traditional solid-state fermentation of vinegar. PMID- 21645818 TI - Diversity of Salmonella isolates using serotyping and multilocus sequence typing. AB - One hundred and twenty-one Salmonella isolates were obtained from food, feed, and live chicken samples derived from 13 countries or regions. In this study, their subtypes were evaluated by serotyping and multilocus sequence typing (MLST), and their genetic profiles were also characterized. It was demonstrated by serotyping on these isolates that 36 various serovars were obtained in this study, of which three serotypes S. Babelsberg, S. Fresno, and S. II were first found in mainland China. Based on Simpson's index of diversity, the serotyping method had a 0.943 discriminatory power. Meanwhile, there were a total of 42 unique sequence types (STs) characterized by MLST, and the discriminatory power of MLST (D = 0.947) was close to that of the serotyping method. In MLST, hisD revealed the highest levels of nucleotide diversity. In addition, ST-92 was the most common ST represented by 16 Salmonella isolates, followed by ST-367 which was represented by 14 isolates. Seven new alleles were identified, which were associated with other alleles and resulted in the assignment of nine new STs. It was concluded from the results that MLST was generally associated with serotype, but not associated with the epidemiological source of the samples, and antimicrobial resistance patterns. PMID- 21645819 TI - Development of real-time PCR methods to quantify patulin-producing molds in food products. AB - Patulin is a mycotoxin produced by different Penicillium and Aspergillus strains isolated from food products. To improve food safety, the presence of patulin producing molds in foods should be quantified. In the present work, two real-time (RTi) PCR protocols based on SYBR Green and TaqMan were developed. Thirty four patulin producers and 28 non-producers strains belonging to different species usually reported in food products were used. The patulin production was tested by mycellar electrokinetic capillary electrophoresis (MECE) and high-pressure liquid chromatography-mass spectrometry (HPLC-MS). A primer pair F-idhtrb/R-idhtrb and the probe IDHprobe were designed from the isoepoxydon dehydrogenase (idh) gene, involved in patulin biosynthesis. The functionality of the developed method was demonstrated by the high linear relationship of the standard curves constructed with the idh gene copy number and Ct values for the different patulin producers tested. The ability to quantify patulin producers of the developed SYBR Green and TaqMan assays in artificially inoculated food samples was successful, with a minimum threshold of 10 conidia g(-1) per reaction. The developed methods quantified with high efficiency fungal load in foods. These RTi-PCR protocols, are proposed to be used to quantify patulin-producing molds in food products and to prevent patulin from entering the food chain. PMID- 21645820 TI - Combinations of High Intensity Light Pulses and Thermosonication for the inactivation of Escherichia coli in orange juice. AB - The non-thermal technologies High Intensity Light Pulses (HILP) and Thermosonication (TS) were applied alone and in combination to study their effect on Escherichia coli inactivation in orange juice. Two different energy settings were chosen in the current study, 'Low' (L) and 'High' (H), being the combinations applied: HILP(L) (4.03 J/cm(2)), HILP(H) (5.1 J/cm(2)), TS(L) (2.8 min residence time at 40 degrees C) and TS(H) (5 min residence time at 50 degrees C). Both the individual technologies and their combinations (HILP&TS and TS&HILP) were studied. Results showed inactivation ranging from 1.10 (TS(H)) to 2.42 (HILP(H)) log cfu/ml for the hurdles when applied individually and from 2.5 (HILP(L)&TS(H)) to 3.93 (HILP(H)&TS(L)) log cfu/ml for the combined treatments. Similar reductions in E. coli populations were achieved in orange juice by all treatment combinations irrespective of the sequence in which they were applied. PMID- 21645821 TI - Use of recA gene sequence analysis for the identification of Staphylococcus equorum strains predominant on dry-cured hams. AB - Spanish dry-cured ham is an uncooked meat product highly appreciated due to its characteristics flavour. In this study, we examined the accuracy of biochemical tests and 16S rDNA sequencing in the identification of 56 staphylococcal strains isolated during industrial Spanish dry-cured ham processes. Important differences were observed comparing genotypic and phenotypic data. Staphylococcus xylosus was the prevalent species identified by biochemical methods (87.5%), however, sequencing of the 16S rDNA resulted in an unambiguous identification of Staphylococcus equorum (73.2%) and Staphylococcus vitulinus (8.9%) strains. Reliable identification of meat staphylococci, mainly among S. xylosus and S. equorum strains could be also achieved by means of recA gene sequence comparison. Two degenerate primers previously described for lactic acid bacteria were used to amplify an internal fragment of the recA gene. This fragment was amplified from twelve staphylococcal type strains representing frequent meat species. The results indicated that recA sequencing is an adequate method to discriminate among meat staphylococci. In addition, S. xylosus and S. equorum strains could be more accurately discriminated by recA sequencing than 16S rDNA or sodA sequencing. The S. equorum sequence diversity showed at the intra-species level by recA gene sequencing confirmed the high heterogeneity described among S. equorum strains. PMID- 21645822 TI - The effect of the number of sample units tested on the precision of microbial colony counts. AB - Studies on the precision of chemical methods of analysis, and the associated 'sampling uncertainty', suggest that analysis of eight replicate sample units (the sample size) is required to ensure adequate analytical precision. The primary purpose of this work was to assess whether these findings are equally applicable in microbiological examination of foods. We examined the effect of sample size on the analytical precision of microbiological data by iteratively 're-sampling without replacement' (SNR). Using both theoretical data sets and colony counts from foods we demonstrate that SNR provides an effective and efficient guide to (a) choosing the number of samples to be examined in order to optimise precision and (b) deciding whether logarithmic transformation of the raw data is appropriate. We also discuss theoretical aspects of the procedure and their impact on the results obtained. PMID- 21645823 TI - Surfome analysis of a wild-type wine Saccharomyces cerevisiae strain. AB - The yeast Saccharomyces cerevisiae, besides being an eukaryotic cell model, plays a fundamental role in the production of fermented foods. In the winemaking industry, yeast cell walls may be involved in numerous processes and contribute substantially to the final chemical and sensorial profiles of wines. Nonetheless, apart from mannoproteins, little is known on the protein components of the yeast cell wall and their changes during the fermentation of must into wine. In this work, we performed a dynamic analysis of the cell surface proteome (surfome) of an autochthonous wine yeast strain (previously selected as a wine fermentation starter) by shaving intact cells with trypsin and identifying tryptic peptides by means of nLC-ESI-LIT-MS/MS. Out of the 42 identified proteins, 16 and 14 were found to be specifically expressed in wine yeast surfome at the beginning and at the end of fermentation, respectively. The molecular functions of these specifically expressed proteins might help in explaining their roles in the cell wall as a response to the alcoholic fermentation-related stresses. Additionally, we provided the identification of 20 new potential cell wall related proteins. Globally, our results might provide new useful data for the selection and characterization of yeast strains to be used in the winemaking industry. PMID- 21645824 TI - Lysozyme as a barrier to growth of Bacillus anthracis strain Sterne in liquid egg white, milk and beef. AB - In this study, we investigated the role of lysozyme on the viability of Bacillus cereus, Bacillus subtilis, Bacillus pumilus and Bacillus anthracis (Sterne) in egg white (EW), ground beef and milk. At 35 degrees C in EW, growth rates (GR) for B. cereus, B. subtilis, B. pumilus and B. anthracis were 0.005, -0.018, 0.028 and -0.029 OD(600)/h, respectively. Heat-treating EW at 55 and 60 degrees C reduced the inactivating effect of EW by 3.1 and 10.5-fold, respectively. Addition of lysozyme (2 mg/ml) to 60 degrees C-treated EW increased the inactivation rate 5.76-fold, indicating involvement of lysozyme in B. anthracis inactivation. B. anthracis inactivation was influenced by pH, as shown by a progressive increase in inactivation rate from 0.25 to -4.42 logs CFU/h over a pH range of 6.0-8.5. Adding 2 mg/ml lysozyme to milk and ground beef also suppressed the growth of B. anthracis 3.3 and 6.5-fold, respectively. These data indicate that lysozyme, as a natural component of EW or potential additive in other foods, could reduce biothreat risks presented by bioterror agents. PMID- 21645825 TI - Prevalence and counts of Salmonella spp. in minimally processed vegetables in Sao Paulo, Brazil. AB - Minimally processed vegetables (MPV) may be important vehicles of Salmonella spp. and cause disease. This study aimed at detecting and enumerating Salmonella spp. in MPV marketed in the city of Sao Paulo, Brazil. A total of 512 samples of MPV packages collected in retail stores were tested for Salmonella spp. and total coliforms and Escherichia coli as indication of the hygienic status. Salmonella spp. was detected in four samples, two using the detection method and two using the counting method, where the results were 8.8 * 10(2) CFU/g and 2.4 * 10(2) CFU/g. The serovars were Salmonella Typhimurium (three samples) and Salmonella enterica subsp. enterica O:47:z4,z23:- (one sample). Fourteen samples (2.7%) presented counts of E. coli above the maximum limit established by the Brazilian regulation for MPV (10(2) CFU/g). Therefore, tightened surveillance and effective intervention strategies are necessary in order to address consumers and governments concerns on safety of MPV. PMID- 21645826 TI - Evaluation of a loop-mediated isothermal amplification assay for rapid and simple detection of Vibrio parahaemolyticus in naturally contaminated seafood samples. AB - We investigated the efficacy of a loop-mediated isothermal amplification (LAMP) assay for rapid screening of seafood samples naturally contaminated with Vibrio parahaemolyticus. A total of 171 seafood samples enriched in alkaline peptone water (APW) were assessed by LAMP assay and conventional culture methods, which consist of a combination of APW enrichment culture and plating onto CHROMagar Vibrio and TCBS agars. Compared with V. parahaemolyticus isolation using the conventional culture test, LAMP results showed 100% (30/30) and 90.8% (128/141) sensitivity and specificity, respectively. The conventional culture test required more than 3 days to isolate and identify V. parahaemolyticus in the APW enrichment culture. In contrast, the LAMP assay was markedly faster, requiring less than 60 min from the beginning of DNA extraction to final detection of V. parahaemolyticus. In total, the LAMP assay required 17-19 h from the beginning of enrichment culture to final determination. This is the first report of the LAMP assay for rapid screening of seafood samples naturally contaminated by V. parahaemolyticus. PMID- 21645827 TI - [Imputability of death to a respiratory occupational disease: a meaningful notion]. PMID- 21645828 TI - [The inspiratory muscle training for all?]. PMID- 21645829 TI - [Medical expert reports in chest disease; the question of imputability of death]. AB - In the course of an investigation, judicial or not, the expert opinion encompasses several questions of a different nature, including the following one " did the patient die of a disease he/she was supposed to suffer from at time of death? " Based on a personal experience over one year in 2008, the goal of this paper is to tackle this question of imputability, which was asked in respect of 12 investigations, including ten of occupational diseases, one of nosocomial infection and one iatrogenic accident. Only two autopsies were carried out; one autopsy refusal was reported. In five out of 12 cases, the imputability of death related to an occupational disease or an iatrogenic accident was considered by the expert to be certain in one case, very probable in two cases, and possible in two cases; in seven out to 12 cases, imputability of death was unlikely, since the cause of death was unknown in two cases, or was not the suggested cause in five cases. The discussion considers several arguments that can help answer this question: evaluation of the vital prognosis of the disease, the importance of the quality of medical records, the contributions and limits of autopsy findings, deaths that result from multiple causes, and the concept of aggravating circumstances. PMID- 21645830 TI - [Inspiratory muscles strength training in recreational athletes]. AB - INTRODUCTION: Respiratory muscles strength and endurance influence athletic performance. Besides conventional spirometry, sniff test, inspiratory and expiratory maximal pressures can directly assess respiratory muscle strength. Respiratory muscles can be train through a device offering inspiratory and expiratory resistance. METHODS: Nineteen subjects aged 18 to 30 years and practicing leisure sport trained inspiratory muscles on Powerbreathe((r)) for eight weeks. Resistance was set at 85% of maximal inspiratory pressure determined during a preliminary session. Evaluation was made trough voluntary and non invasive methods on Macro 5000((r)) (PI max, PE max and sniff test). RESULTS: An increase of 21.77% of the maximum inspiratory pressure, 17% of the maximum expiratory pressure and 18% of the sniff test are recorded after eight weeks of training. CONCLUSIONS: A specific training of inspiratory muscles (Powerbreathe((r)) Sports performance) increases the power of these muscles (voluntary and non-invasive tests). PMID- 21645831 TI - [Value of tomotherapy in malignant pleural mesothelioma: first clinical results]. AB - INTRODUCTION: There is little clinical data about the place of helicoidal tomotherapy (HT) in the treatment of malignant pleural mesothelioma (MPM). This new form of intensity modulated radiotherapy (IMRT) has great theoretical advantages in large and complex volumes when compared to "traditional" forms of radiotherapy. PATIENTS AND METHODS: Fourteen patients diagnosed with MPM received adjuvant radiotherapy by HT. The patients were treated at the Curie Institute and the Rene Gauducheau Centre, starting in August 12007. All patients had a complete initial staging, an extrapleural pneumonectomy (EPP), and a minimum follow-up of six months. The median dose prescribed to the surgical cavity was 50 Gy (48-54 Gy) in 2 Gy (1.80-2.07) fractions. High dose regions received concomitant 57 Gy (54-69 Gy) in 2.16 Gy (2.00-2.30 Gy) fractions. RESULTS: Median follow-up was 12.6 months after ending HT. Seven patients received neoadjuvant chemotherapy (cisplatin or carboplatin, and pemetrexed). Eight patients were staged pT3 and five were staged pN1-2. HT was well tolerated. Two patients had suspected G5 radiation pneumonitis within 6 months of ending HT. Of the 12 patients who survived treatment, six relapsed (in average 5.1 months after HT): distant. Four relapses were distant; two relapses were both local and distant. Three patients died after their initial relapse. After initial diagnosis, the median survival was 18.4 months. A learning curve was observed in the optimization of the dosimetric parameters. CONCLUSION: Helicoidal tomotherapy is a reliable, quite well tolerated, and efficient way of treating MPM patients after an EPP. PMID- 21645832 TI - [Positron emission tomography with 18F-FDG and cancer response to chemotherapy]. AB - INTRODUCTION: Few data are available about the predictive value of FDG-PET-CT in the evaluation of the response to chemotherapy of patients with advanced NSCLC and its impact on subsequent survival. METHODS: A retrospective study of patients with advanced NSCLC who underwent a FDG-PET-CT before treatment and after three cycles of first-line chemotherapy. Morphological and metabolic responses were assessed respectively using RECIST/OMS and EORTC criteria. The relation between response and survival was analysed through Cox regression models. RESULTS: Fifty nine patients were included in the study (stage IIIA/IIIB/IV: 9/11/39). Median survival was 40 weeks (44 deaths observed). The evaluation of treatment response (morphological or metabolic, taken alone or combined) in terms of survival failed to identify any difference between responders and patients with stable disease. Only patients with progressive disease had a significantly shorter survival. The negative predictive value of the metabolic response for the morphological response is 0.90 and that of metabolic progression for morphological progression is 0.98. CONCLUSION: Only progressive disease differs significantly from other types of response with a more sensitive and earlier detection by PET-CT. PMID- 21645833 TI - [Patients'illness perceptions and adherence to treatment with inhaled corticosteroids in asthma]. AB - INTRODUCTION: Regular use of inhaled corticosteroids as preventive treatment of asthma is an integral part of management but remains inadequate among adults. Studying the perceptions of illness and treatment beliefs is one way to understand the patient's adherence to medication. METHOD: A systematic review was performed of empirical studies in adults published between 1999 and 2009, and indexed in the Pubmed, PsycInfo and Scopus databases. We investigated the associations between (1) perceptions of asthma and treatment beliefs and (2) adherence to inhaled corticosteroids. Eighteen articles meet these criteria. RESULTS: Perception of the chronicity of asthma and its consequences on daily life, as well as the concept that it is necessary to continue treatment in the absence of symptoms, are associated with better adherence. On the contrary, fear of side effects and the belief that treatment is ineffective in controlling symptoms, are associated with poor adherence. CONCLUSION: Patients' perceptions of asthma and inhaled corticosteroids are predictors of adherence to treatment. The identification and discussion of these issues is an essential part of building a therapeutic relationship that facilitates adherence. PMID- 21645834 TI - [Tobacco and marijuana use in adolescent students in Noumea]. AB - INTRODUCTION: New Caledonia is situated in the western South Pacific 20000 km from France. In this Overseas Territory (pays d'outre-mer [POM]) the prevalence of tobacco smoking is very high and estimated at 30% among men and 34% among women. Experimenting with cannabis is also very widespread. The incidence of chronic respiratory illness and lung cancer remains high in the Territory. Modern laws protecting non-smokers still do not exist. This study aims at describing the behaviour of young people from 14 to 18 years old, at school in the public sector of Noumea, with respect to tobacco and marijuana consumption. This survey examines the way this behaviour varies according to age, sex, community, place of residence and socioeconomic conditions, in order to identify the most exposed groups. Finally, we study the effect of this consumption on the respiratory health of young school people. METHODS: The enquiry was undertaken from May 4th to 15th 2009 in 16 classes drawn randomly from the colleges and secondary schools of Noumea (439 pupils). The survey consisted of an anonymous questionnaire containing 48 questions grouped into five subjects: a sociodemographic description of the subject and his/her family, the family lifestyle, tobacco consumption, marijuana consumption and a respiratory questionnaire. The completion of the questionnaires took, on average, 25 minutes and took place in class in the presence of the doctor undertaking the survey. The data were analysed with Ethnos-4 software. The analysis was made in two stages: a descriptive study after uni- and bivariate analysis and an analytical study to identify the risk factors of the addictive practices. The statistical tests used were Pearson's chi(2) test and the Hosmer-Lemeshow test for calculation of odds ratio. RESULTS: The participation rate was 95.2% (n = 415). The sample was predominantly female (56.6%) and 31% of the pupils were less than 16 years old. The number of tobacco smokers was considerable at 41.1%, 27.3% were regular daily smokers including 38.5% who smoked more than six cigarettes a day. The results were dependent on sex (female predominence) but independent of ethnic origin and socioeconomic factors. The number of cannabis smokers was 48% among whom 32% were regular consumers. The daily smokers were uniquely boys and the custom was more frequent among the Melanesian population. Only 11% of the pupils were worried about their consumption and, among them, only the regular smokers were more at risk of bronchial infection. CONCLUSIONS: In New Caledonia, tobacco consumption levels are higher than those in developed countries. We did not find a falling trend compared with previous data. The use of cannabis remains very widespread at an early age, with a male predominance for daily consumption. For both, these addictions the perception of the risk and the desire to stop are weak. The population studied is representative of teenagers in the public sector schools in Grand-Noumea but probably does not reflect the situation on the whole territory. The results obtained suggest the value of a study of the whole of New Caledonia, the eventual purpose being to guide the public health authorities towards policies that help the young people of the country. PMID- 21645835 TI - [Clinical characteristics of resected bronchial hamartoma. Study of seven cases]. AB - INTRODUCTION: Endobronchial hamartoma is a benign tumour derived from peribronchial mesenchymal tissue. It is a form of intrapulmonary hamartoma and can cause irreversible pulmonary destruction due to bronchial obstruction. Early diagnosis and treatment is very important and endoscopic treatment is usually the first choice. In cases in which prolonged bronchial obstruction has produced irreversible lung destruction surgical, resection of the tumour is necessary. The aim of this study is to describe the clinicopathological characteristics of resected endobronchial hamartoma. PATIENTS AND METHODS: Seven cases of endobronchial hamartoma, diagnosed in our institution between January 1995 and December 2009, were reviewed retrospectively. RESULTS: The patient population consisted of four males and three females with mean age of 53.7 years (41-68 years). The most frequent clinical presentation was obstructive pneumonia. Endoscopy and imaging showed an endobronchial mass in most cases. Surgical treatment consisted of atypical resection in one case, lobectomy in five cases and pneumonectomy in one case. Tumour size ranged between 0.5 and 3.5 cm. The diagnosis was confirmed in all cases by histological examination. The outcome was favorable in all cases. CONCLUSION: Endoscopic techniques are effective for the diagnosis and treatment of endobronchial hamartochondroma. Treatment should be prompt to prevent irreversible lung damage due to chronic obstruction and suppuration, in which case, pulmonary resection may be necessary. PMID- 21645836 TI - [Effect of tinzaparin on survival in non-small-cell lung cancer after surgery. TILT: tinzaparin in lung tumours]. AB - BACKGROUND: Experimental and clinical findings suggest that low molecular-weight heparins may improve overall survival in patients with cancer. The evidence is still limited and additional studies are needed to confirm these preliminary findings. METHODS: Patients with completely resected stage I, II or IIIA (T3N1) histologically confirmed non-small-cell lung cancer will be included in a prospective, controlled, randomized, multicenter open trial. Patients in the control group will receive usual postoperative care including chemotherapy when indicated. Patients in the experimental group will receive tinzaparin given subcutaneously as a daily 100 IU/kg dose for 90 days along with usual postoperative care. Patients will be followed-up for three to eight years. Main end-point is the overall survival. Five hundred and fifty patients are needed to demonstrate a 10% absolute increase in survival in the experimental group. EXPECTED RESULTS: A 10% absolute increase in the survival rate is expected in the patients receiving tinzaparin. PMID- 21645837 TI - [A rare cause of miliary]. PMID- 21645838 TI - [Atypical unilateral insterstitial disease]. PMID- 21645839 TI - ["On and off" hypoxaemia]. AB - INTRODUCTION: A right to left shunt caused by a patent foramen ovale (PFO) must be considered when patients present with symptoms of platypnoea-orthodeoxia. The most useful investigation is saline contrast transthoracic or transoesophageal echocardiography. CASE REPORT: We report a case of an eighty-year-old woman with the platypnoea-orthodeoxia syndrome, but without signs of a right to left shunt caused by a PFO. (99m)Tc-macroaggregated albumin lung scintigraphy and saline contrast transthoracic echocardiography were considered normal in the supine position. The clinical suspicion of PFO was so strong that the examinations were repeated in the upright position. This revealed a systemic uptake of the isotope on lung scintigraphy, confirmed by saline contrast echocardiography. The atrial septal defect was due to displacement of the interatrial septum by an aneurysm of the ascending aortic. In the upright position blood flowed directly from the inferior cava vena through a PFO into the left atrium. CONCLUSION: The diagnosis of PFO may be difficult and it is important to repeat saline contrast echocardiography in the upright position when it is negative supine. It is possible to confirm the diagnosis by contrast infusion through the femoral veins. PMID- 21645840 TI - [Congenital bronchial atresia in adults]. AB - Congenital bronchial atresia is a rare congenital obliteration of a segmental or lobar bronchus resulting in distension of the corresponding parenchyma. It is seldom diagnosed in the adult. It may lead to infectious complications and, in the long term, to damage to the adjacent lung parenchyma. A surgical resection is necessary and it can be achieved by thoracoscopy. We report a recent series of six patients. PMID- 21645841 TI - [Pulmonary hamartoma with lipomatous form]. AB - We report of the case of a 41-year-old patient, who had previously undergone thoracic surgery at the age of 16 for a single giant emphysematous bulla. The CT scan showed an abnormal middle mediastinal lesion containing an aerated cystic areas and areas of fat density. The patient underwent surgery and a well-defined mass was found between the mediastinum and the pulmonary hilum, which was able to be completely resected. Microscopic examination disclosed a composite tumour containing a bronchogenic cyst, a benign lipoma and two hamartochondromas. As no similar case had been reported previously, it was difficult to assert the nature of the lesion, which has been labelled as a "hamartoma". The main diagnoses which should be considered when an intra pulmonary fat density mass is disclosed, are discussed. PMID- 21645842 TI - [Rare primary chest wall sarcoma: the synovialosarcoma]. AB - INTRODUCTION: Malignant primary tumours occurring in the thorax encompass a large group of tumours which may arise from the lung, mediastinal structures, the pleura or the chest wall. OBSERVATION: We report the case of a 37 year old patient, who presented with left sided chest pain. On clinical examination a right sided chest wall mass was identified. Chest X Ray showed a left sided upper mediastinal opacity, associated with a left sided pleural opacity. Thoracic CT scan revealed a large mass arising from the chest wall and infiltrating the mediastinum associated with a second chest wall mass at the level of the 8(th) and 9(th) right ribs. The biopsy of the chest wall mass revealed it to be a parietal synovialosarcoma. The patient responded to chemotherapy based on ifosfamid and doxorubicin as well as mediastino-pulmonary radiotherapy. There was an improvement in the patient's clinical and radiological state but the patient died by pulmonary embolism after the 3(rd) cause of treatment. CONCLUSION: Chest wall synovialosarcoma has a poor prognosis, however, its chemosensitivity means that treatment may initially be effective. PMID- 21645843 TI - [Multiple pulmonary hydatid disease with cardiovascular localisation]. AB - Multiple thoracic hydatid disease is rare. Cardiovascular localisation is exceptional. Clinical, radiological and even electrocardiographic signs are not specific. The diagnosis is often difficult. We report a case of multiple pulmonary hydatid disease associated to a hydatid cyst of the infundibulum of the pulmonary artery. This is presented in a young man, aged 21 years, in good physical health, with recurrent minimal haemoptysis dating back two years ago. The authors emphasise, throughout this case history, the difficulties of diagnosis and treatment of multiple hydatid cysts especially with cardiovascular localisation. PMID- 21645844 TI - [Enigmatic evolution of an association of pulmonary tuberculosis and amyloidosis]. AB - INTRODUCTION: Amyloidosis is characterized by tissue deposits of amyloid material. Secondary amyloidosis can occur as a sequel to pulmonary tuberculosis over a relatively long period. However, this was not the case with our patient. Subsequently we conducted a literature review to try to explain the unusual course of AL amyloidosis in our patient. CASE REPORT: A 36- year-old patient was admitted to our department for investigation of haemoptysis. A diagnosis of primary pulmonary tuberculosis was made and antituberculous treatment was started. On the second day of treatment, a haematoma appeared on the sole of the right foot, which spread down to the toes during the following days. Renal investigations showed a 24h proteinuria of 9 g/L and serum protein electrophoresis revealed an albumin level of 11.8 g/L. A diagnosis of nephrotic syndrome was made. A renal biopsy was indicated but this was not possible on account of a marked worsening of the patient's condition after 14 days of treatment. The patient's level of consciousness deteriorated and he was transferred to the intensive care unit for ventilation. He died 48 hours later. Post-mortem histological examination of pulmonary and cutaneous tissue revealed AL amyloid deposits. CONCLUSION: In view of the association of active pulmonary tuberculosis and a pulmonary localisation of amyloidosis, a causal relationship is not definite. Coexistence of active pulmonary tuberculosis and primary amyloidosis must also be considered, particularly as the immunohistochemical characterisation revealed AL amyloidosis. PMID- 21645845 TI - [A conus medullaris tuberculoma: a case report and literature review]. AB - BACKGROUND: Tuberculosis affecting the central nervous system is well recognized, but only rarely localizes to the medullary conus. OBSERVATION: We report the case of a 69 year old man who was admitted to our unit with cauda equina syndrome. The MRI demonstrated ring-enhanced necrotizing lesions involving the medullary conus, the cervical cord and the brain. His chest CT scan showed a miliary infiltrate. The clinical presentation was associated with an inappropriate secretion of antidiuretic hormone. Quadruple antituberculous therapy was initiated, with corticosteroids in the initial phase of the treatment. Evolution was favorable, and follow-up MRI imaging demonstrated complete resolution of the cervical cord and brain lesions. PMID- 21645846 TI - Unveiled alpha-neurexins take center stage. AB - Neurexins are presynaptic transmembrane proteins that play an essential role in synapse function. The crystal structures of alpha-neurexin extracellular domains (Chen et al., 2011; Miller et al., 2011) provide important insights into their conformational freedom and their putative spatial arrangement with binding partners in the synaptic cleft. PMID- 21645847 TI - Recombining DNA by protein swivels. AB - Two new reports on serine recombinases, one of a crystal snapshot in an alternate rotational conformer poised for DNA cleavage (Keenholtz et al., 2011), and a second employing single-DNA molecule approaches (Bai et al., 2011), provide strong support for the subunit rotation model for exchanging DNA strands. PMID- 21645848 TI - Structural changes underlying allostery in group II chaperonins. AB - The ATP-dependence of folding chamber closure in the 16-subunit homo-oligomeric chaperonin from archaea Methanococcus maripaludis (Mm-cpn) has been studied by single particle cryo-electron microscopy (Zhang et al., 2011). ATP binding alone causes a rigid body rotation of ~45 degrees and slight closure of the cavity, but full closure requires ATP hydrolysis. PMID- 21645849 TI - Improved technologies now routinely provide protein NMR structures useful for molecular replacement. AB - Molecular replacement (MR) is widely used for addressing the phase problem in X ray crystallography. Historically, crystallographers have had limited success using NMR structures as MR search models. Here, we report a comprehensive investigation of the utility of protein NMR ensembles as MR search models, using data for 25 pairs of X-ray and NMR structures solved and refined using modern NMR methods. Starting from NMR ensembles prepared by an improved protocol, FindCore, correct MR solutions were obtained for 22 targets. Based on these solutions, automatic model rebuilding could be done successfully. Rosetta refinement of NMR structures provided MR solutions for another two proteins. We also demonstrate that such properly prepared NMR ensembles and X-ray crystal structures have similar performance when used as MR search models for homologous structures, particularly for targets with sequence identity >40%. PMID- 21645850 TI - The deletion of exon 3 in the cardiac ryanodine receptor is rescued by beta strand switching. AB - Mutations in the cardiac Ryanodine Receptor (RYR2) are linked to triggered arrhythmias. Removal of exon 3 results in a severe form of catecholaminergic polymorphic ventricular tachycardia (CPVT). This exon encodes secondary structure elements that are crucial for folding of the N-terminal domain (NTD), raising the question of why the deletion is neither lethal nor confers a loss of function. We determined the 2.3 A crystal structure of the NTD lacking exon 3. The removal causes a structural rescue whereby a flexible loop inserts itself into the beta trefoil domain and increases thermal stability. The exon 3 deletion is not tolerated in the corresponding RYR1 domain. The rescue shows a novel mechanism by which RYR2 channels can adjust their Ca2+ release properties through altering the structure of the NTD. Despite the rescue, the deletion affects interfaces with other RYR2 domains. We propose that relative movement of the NTD is allosterically coupled to the pore region. PMID- 21645851 TI - Structural basis for catalytic activation of a serine recombinase. AB - Sin resolvase is a site-specific serine recombinase that is normally controlled by a complex regulatory mechanism. A single mutation, Q115R, allows the enzyme to bypass the entire regulatory apparatus, such that no accessory proteins or DNA sites are required. Here, we present a 1.86 A crystal structure of the Sin Q115R catalytic domain, in a tetrameric arrangement stabilized by an interaction between Arg115 residues on neighboring subunits. The subunits have undergone significant conformational changes from the inactive dimeric state previously reported. The structure provides a new high-resolution view of a serine recombinase active site that is apparently fully assembled, suggesting roles for the conserved active site residues. The structure also suggests how the dimer tetramer transition is coupled to assembly of the active site. The tetramer is captured in a different rotational substate than that seen in previous hyperactive serine recombinase structures, and unbroken crossover site DNA can be readily modeled into its active sites. PMID- 21645852 TI - Domain orientation in the N-Terminal PDZ tandem from PSD-95 is maintained in the full-length protein. AB - Tandem PDZ domains have been suggested to form structurally independent supramodules. However, dissimilarity between crystallography and NMR models emphasize their malleable conformation. Studies in full-length scaffold proteins are needed to examine the effect of tertiary interactions within their native context. Using single-molecule fluorescence to characterize the N-terminal PDZ tandem in PSD-95, we provide the first direct evidence that PDZ tandems can be structurally independent within a full-length scaffold protein. Molecular refinement using our data converged on a single structure with an antiparallel alignment of the ligand-binding sites. Devoid of interaction partners, single molecule conditions captured PSD-95 in its unbound, ground state. Interactions between PDZ domains could not be detected while fluctuation correlation spectroscopy showed that other conformations are dynamically sampled. We conclude that ultra-weak interactions stabilize the conformation providing a "low-relief" energy landscape that allows the domain orientation to be flipped by environmental interactions. PMID- 21645853 TI - Implications for damage recognition during Dpo4-mediated mutagenic bypass of m1G and m3C lesions. AB - DNA is susceptible to alkylation damage by a number of environmental agents that modify the Watson-Crick edge of the bases. Such lesions, if not repaired, may be bypassed by Y-family DNA polymerases. The bypass polymerase Dpo4 is strongly inhibited by 1-methylguanine (m1G) and 3-methylcytosine (m3C), with nucleotide incorporation opposite these lesions being predominantly mutagenic. Further, extension after insertion of both correct and incorrect bases, introduces additional base substitution and deletion errors. Crystal structures of the Dpo4 ternary extension complexes with correct and mismatched 3'-terminal primer bases opposite the lesions reveal that both m1G and m3C remain positioned within the DNA template/primer helix. However, both correct and incorrect pairing partners exhibit pronounced primer terminal nucleotide distortion, being primarily evicted from the DNA helix when opposite m1G or misaligned when pairing with m3C. Our studies provide insights into mechanisms related to hindered and mutagenic bypass of methylated lesions and models associated with damage recognition by repair demethylases. PMID- 21645854 TI - Hierarchical binding of cofactors to the AAA ATPase p97. AB - The hexameric AAA ATPase p97 is involved in several human proteinopathies and mediates ubiquitin-dependent protein degradation among other essential cellular processes. Via its N-terminal domain (N domain), p97 interacts with multiple regulatory cofactors including the UFD1/NPL4 heterodimer and members of the "ubiquitin regulatory X" (UBX) domain protein family; however, the principles governing cofactor selectivity remain to be deciphered. Our crystal structure of the FAS-associated factor 1 (FAF1)UBX domain in complex with the p97N domain reveals that the signature Phe-Pro-Arg motif known to be crucial for interactions of UBX domains with p97 adopts a cis-proline configuration, in contrast to a cis trans mixture we derive for the isolated FAF1UBX domain. Biochemical studies confirm that binding critically depends on a proline at this position. Furthermore, we observe that the UBX proteins FAF1 and UBXD7 only bind to p97 UFD1/NPL4, but not free p97, thus demonstrating for the first time a hierarchy in p97-cofactor interactions. PMID- 21645855 TI - A smoothed backbone-dependent rotamer library for proteins derived from adaptive kernel density estimates and regressions. AB - Rotamer libraries are used in protein structure determination, prediction, and design. The backbone-dependent rotamer library consists of rotamer frequencies, mean dihedral angles, and variances as a function of the backbone dihedral angles. Structure prediction and design methods that employ backbone flexibility would strongly benefit from smoothly varying probabilities and angles. A new version of the backbone-dependent rotamer library has been developed using adaptive kernel density estimates for the rotamer frequencies and adaptive kernel regression for the mean dihedral angles and variances. This formulation allows for evaluation of the rotamer probabilities, mean angles, and variances as a smooth and continuous function of phi and psi. Continuous probability density estimates for the nonrotameric degrees of freedom of amides, carboxylates, and aromatic side chains have been modeled as a function of the backbone dihedrals and rotamers of the remaining degrees of freedom. New backbone-dependent rotamer libraries at varying levels of smoothing are available from http://dunbrack.fccc.edu. PMID- 21645856 TI - Relative solvent accessible surface area predicts protein conformational changes upon binding. AB - Protein interactions are often accompanied by significant changes in conformation. We have analyzed the relationships between protein structures and the conformational changes they undergo upon binding. Based upon this, we introduce a simple measure, the relative solvent accessible surface area, which can be used to predict the magnitude of binding-induced conformational changes from the structures of either monomeric proteins or bound subunits. Applying this to a large set of protein complexes suggests that large conformational changes upon binding are common. In addition, we observe considerable enrichment of intrinsically disordered sequences in proteins predicted to undergo large conformational changes. Finally, we demonstrate that the relative solvent accessible surface area of monomeric proteins can be used as a simple proxy for protein flexibility. This reveals a powerful connection between the flexibility of unbound proteins and their binding-induced conformational changes, consistent with the conformational selection model of molecular recognition. PMID- 21645857 TI - The cap-binding translation initiation factor, eIF4E, binds a pseudoknot in a viral cap-independent translation element. AB - Eukaryotic initiation factor eIF4E performs a key early step in translation by specifically recognizing the m7GpppN cap structure at the 5' end of cellular mRNAs. Many viral mRNAs lack a 5' cap and thus bypass eIF4E. In contrast, we reported a cap-independent translation element (PTE) in Pea enation mosaic virus RNA2 that binds and requires eIF4E for translation initiation. To understand how this uncapped RNA is bound tightly by eIF4E, we employ SHAPE probing, phylogenetic comparisons with new PTEs discovered in panico- and carmoviruses, footprinting of the eIF4E binding site, and 3D RNA modeling using NAST, MC-Fold, and MC-Sym to predict a compact, 3D structure of the RNA. We propose that the cap binding pocket of eIF4E clamps around a pseudoknot, placing a highly SHAPE reactive guanosine in the pocket in place of the normal m7GpppN cap. This reveals a new mechanism of mRNA recognition by eIF4E. PMID- 21645858 TI - A systematic study of the energetics involved in structural changes upon association and connectivity in protein interaction networks. AB - The study of protein binding mechanisms is a major topic of research in structural biology. Here, we implement a combination of metrics to systematically assess the cost of backbone conformational changes that protein domains undergo upon association. Through the analyses of 2090 unique unbound -> bound transitions, from over 12,000 structures, we show that two-thirds of these proteins do not suffer significant structural changes upon binding, and could thus fit the lock-and-key model well. Among the remaining proteins, one-third explores the bound conformation in the unbound state (conformational selection model) and, while most transitions are possible from an energetic perspective, a few do require external help to break the thermodynamic barrier (induced fit model). We also analyze the relationship between conformational transitions and protein connectivity, finding that, in general, domains interacting with many partners undergo smaller changes upon association, and are less likely to freely explore larger conformational changes. PMID- 21645859 TI - Fine details of IGF-1R activation, inhibition, and asymmetry determined by associated hydrogen /deuterium-exchange and peptide mass mapping. AB - The structural features of the asymmetric activated states of the insulin receptor family are still poorly understood. We investigated hydrogen/deuterium (H/D)-exchange within the extracellular domain of the type-I insulin-like growth factor receptor (IGF-1R) in the absence and presence of IGF-1 (active state) and in the presence of antibody inhibitors (inactive state). Near complete coverage of the 210 kDa receptor sequence was obtained by mass mapping of proteolytically derived peptides at all H/D-exchange time points. The data provide details regarding solvent exposure and dynamics across the extracellular region as well as conformational changes induced by activation or inactivation. Multiple peptides, distant in structure, individually demonstrated two distinct H/D exchange rates, suggesting that each of these peptides exists in two separate environments in IGF-1R. The dual-exchange behavior of these peptides was enhanced on ligand binding and eliminated on inhibitor binding, clearly associating these regions with active state asymmetry and enabling them to serve as reporters of receptor activity. PMID- 21645860 TI - Introduction: current controversies in rectal cancer. PMID- 21645861 TI - Rectal cancer: primary staging and assessment after chemoradiotherapy. AB - Rectal cancer staging is based on 2 principles. The first is an anatomic definition of the tumor allowing for surgical planning. The second is prognostic stage grouping. A given prognostic stage carries different risks of both local and distant recurrence, a selective and tailored approach to preoperative therapy is appropriate. Increasingly, selective approaches enable an overall reduction in morbidity from overtreatment, while allowing aggressive treatment of high-risk patients. Therefore, the aim of preoperative staging is to accurately and reproducibly differentiate between good and poor prognosis tumors. In the preoperative setting, superficial and flat rectal cancers are probably best initially staged using endoscopic ultrasound, and where available magnetic resonance imaging is used for all other rectal cancers because of its proven high sensitivity and specificity in identifying poor-risk patients based on circumferential margin status, the depth of extramural spread, extramural venous invasion, and nodal status. Restaging after neoadjuvant therapy is a challenge to all modalities because of radiation-induced changes, namely fibrosis, edema, inflammation, and necrosis. However, emerging data suggest that reassessment using a combination of high-resolution magnetic resonance imaging, diffusion weighted imaging, and positron emission tomography/computed tomography scanning may help to provide valuable prognostic information before definitive surgery. PMID- 21645862 TI - Local excision: is it an adequate substitute for radical resection in T1/T2 patients? AB - Local excision (LE) was historically developed to palliate patients with rectal adenocarcinoma who either are medically unfit or have adamantly refused to undergo transabdominal standard resection (SR) procedures. Over the years, the tradeoffs between the oncologic benefit and adverse functional sequelae associated with SR procedures have been increasingly recognized. In parallel, there has been growing interest in considering LE as an alternative to SR in select patients with early-stage disease. However, concerns regarding its oncologic adequacy remain. These concerns relate to the adequacy of tumor resection, the removal of mesorectal disease, the accuracy of preoperative selection, and the use of adjunctive treatment modalities. Evolving strategies that aim at improving the oncologic outcomes of LE for stage I T1/T2 rectal cancers include adoption of transanal endoscopic microsurgery and the addition of non-surgical modalities. Current evidence surrounding these approaches is examined to provide a basis for an informed discussion with patients. Key factors to be considered in formulating the treatment plan for an individual patient with T1/T2 rectal cancer are summarized. PMID- 21645863 TI - How low is low? Evolving approaches to sphincter-sparing resection techniques. AB - Although advances in rectal cancer staging may ultimately be accurate enough to reliably exclude disease outside the rectal wall (thereby allowing local approaches to be more widely and safely applied) and advances in the use of neoadjuvant chemo- and radiation therapy may ultimately produce more "complete responders" that can be accurately identified and spared surgery altogether, as it stands, radical resection forms the basis of curative treatment for rectal cancer. However, the concepts that guide the surgeon in choosing the optimal approach in radical resection are changing. In the past, the decision as to how to proceed surgically with radical resection was based primarily on the level of the tumor above the anal verge or anorectal ring. The issue was primarily "How low is the tumor?" and "Is the distal margin safe?" A more modern approach focuses attention on achieving a negative circumferential margin despite what historically may seem to be a very minimal distal margin, the current issue is not "How low is the tumor?" so much as it is "How deep does the tumor go?". This shift in focus has been a major impetus in the evolution of sphincter sparing resection techniques. PMID- 21645864 TI - Neoadjuvant chemotherapy alone for early-stage rectal cancer: an evolving paradigm? AB - Current management of early-stage rectal cancer comprises combinations of surgery, radiotherapy, and chemotherapy, with the presence or absence of several validated high-risk features determining which treatment modalities will be used and the order of administration. In high-risk individuals, most centers have adopted neoadjuvant combined chemotherapy and radiotherapy followed by surgery as the initial approach. However, long-term toxicity, limited survival gains, and high rates of distant failure have called this approach into question, with early data suggesting that neoadjuvant chemotherapy alone may be feasible in selected patient groups. This review discusses the current data and feasibility of managing early stage rectal cancer with neoadjuvant chemotherapy before surgical resection. PMID- 21645865 TI - Beyond 5-fluorouracil: the emerging role of newer chemotherapeutics and targeted agents with radiation therapy. AB - The management of rectal cancer has undergone significant evolution with advances in surgery, radiation therapy, and chemotherapy. These advances have translated into improved rates of local control, survival, and quality of life. More recently, the integration of newer chemotherapeutic and targeted agents in patients with advanced colorectal cancer have led to further improvements in disease-free and overall survival. These agents are now being studied with radiation therapy in the neoadjuvant therapy of rectal cancer. PMID- 21645866 TI - T3N0 rectal cancer: radiation for all? AB - The optimal oncologic management for patients with T3N0 rectal cancer is currently controversial. Patients with pathologic T3N0 disease may have an "intermediate" risk of disease recurrence, suggesting that perhaps trimodality therapy may not be indicated for all patients. Adverse prognostic features, including a greater depth of perirectal fat invasion, poor tumor differentiation, the presence of lymphovascular invasion, abnormally elevated pretreatment carcinoembryonic antigen levels (>5 ng/mL), circumferential margin involvement, and a low-lying position may identify T3N0 patients at high risk for local recurrence who may benefit from the addition of radiation therapy. However, recent randomized data suggest an improvement in local control and disease-free survival with preoperative radiation therapy compared with selective postoperative radiation therapy in all patient subgroups, arguing in favor of routine preoperative therapy. Additionally, rates of clinical understaging may exceed 20%, representing the percentage of patients who would require the delivery of postoperative radiotherapy with its associated sequelae. Future prospective randomized studies of T3N0 patients with upfront stratification by known prognostic factors and studies evaluating the molecular profile of rectal cancers hold the promise of better classifying patients at high risk of local and systemic recurrence, and thus, in need of adjuvant radiation and chemotherapy. PMID- 21645867 TI - Point: short-course radiation therapy is preferable in the neoadjuvant treatment of rectal cancer. AB - There are 2 types of neoadjuvant radiation regimens accepted as standard for resectable rectal cancer: short-course (5 * 5 Gy) radiation therapy alone with immediate surgery and long-course combined chemoradiation therapy with delayed surgery. A Polish randomized study (n = 312) and an Australian randomized study (n = 326) compared these 2 schedules. Both trials showed a lower rate of early adverse effects using a short-course radiation regimen and no differences in long term oncologic outcomes and late toxicity rates between groups. The small number of fractions makes short-course radiation less expensive and more convenient than chemoradiation therapy. These facts indicate that short-course radiation is preferable to chemoradiation for resectable cancers. Additionally, short-course preoperative radiation with a long interval to surgery is a valuable option for patients unfit for chemotherapy, with unresectable cancer or with a small tumor that is amenable to local excision. Moreover, short-course radiation enables the intensification of both radiotherapy and chemotherapy in patients with metastatic rectal cancer with potentially resectable synchronous metastatic disease. PMID- 21645868 TI - Counterpoint: long-course chemoradiation is preferable in the neoadjuvant treatment of rectal cancer. AB - There are 2 approaches to preoperative therapy. Short-course (25 Gy in 5 fractions) radiation and long-course (50.4 Gy in 28 fractions) radiation combined with chemotherapy (CMT). Although short-course radiation therapy is used in some European countries, it is not favored in all European countries or North America. Unlike long-course CMT, it cannot be safely combined with adequate doses of systemic concurrent chemotherapy, and, as currently designed, it does not increase sphincter preservation. Long-course CMT remains the preferred regimen for cT3 and/or node-positive disease. With parallel advances in staging, surgery, systemic therapy, and molecular markers, more selective approaches are being investigated. PMID- 21645869 TI - Nonoperative approaches to rectal cancer: a critical evaluation. AB - A neoadjuvant multimodality approach with chemoradiation therapy (CRT) is the preferred treatment strategy for most distal rectal cancers. Significant downstaging and complete pathologic response may develop after this strategy, and there is still controversy regarding the management of these patients. In this setting, a nonoperative approach has been suggested in select patients with complete clinical response after thorough clinical, endoscopic, and radiologic assessment. However, the assessment of these patients is not straightforward and remains complex. Available data regarding this approach are limited to a single institution's experience from retrospective analyses. Standardization of the assessment of tumor response and the development of radiological/molecular tools may clarify the role of no immediate surgery in patients with complete clinical response after neoadjuvant CRT. Advances in radiation and medical oncology could potentially lead to significant improvements in complete tumor regression rates, leading to an increase in importance of a minimally invasive approach in patients with rectal cancer. PMID- 21645870 TI - beta-Adrenergic signaling and response to pressure overload in transgenic mice with cardiac-specific overexpression of inducible NO synthase. AB - The role of iNOS induction in the context of cardiac hypertrophy and heart failure is still not fully understood. We have used transgenic mice with cardiac specific overexpression of iNOS (tg-iNOS) to investigate the consequences of high level NO formation on cardiac function in vivo and the response to chronic pressure overload. Conductance manometry was used to analyze cardiac function of wild type (WT) and tg-iNOS mice under basal conditions and beta-adrenergic stimulation. To investigate the influence of iNOS on cardiac function in hypertrophied hearts, transversal aortic constriction was performed. Despite a high level of cardiac NO formation tg-iNOS mice showed almost normal LV function under basal conditions. The cardiac response to beta-adrenergic stimulation, however, was completely abolished. Acute NOS inhibition led to an instantaneous recovery of the inotropic response to catecholamines in tg-iNOS mice. Chronic pressure overload induced a similar extent of cardiac hypertrophy in WT and tg iNOS hearts. LV function, however, was more compromised in tg-iNOS hearts as revealed by a decreased contractility and cardiac output. IN CONCLUSION: a high level of cardiac NO formation does not induce heart failure per se but severely enhances the functional depression in response to pressure overload. This effect could be due to the tonic impairment of the cardiac beta-adrenergic response. PMID- 21645871 TI - Compromised sleep quality and low GPA among college students who use prescription stimulants nonmedically. PMID- 21645872 TI - Arm/shoulder problems and insomnia symptoms in breast cancer survivors: cross sectional, controlled and longitudinal observations. AB - OBJECTIVE: In breast cancer survivors (BCSs) the relation between insomnia symptoms and arm/shoulder problems has hardly been investigated. In cross sectional and longitudinal designs we examined this association in BCSs and in comparison to age-matched controls from the general population. METHODS: Our cross-sectional sample consisted of 337 BCSs stage II/III studied in 2004 at a median of 3.9 years after surgery combined with adjuvant radiotherapy and cytostatics/hormones. In 2007 248 (74%) BCSs were re-examined (median 2.5 years later). The responses of the 2004 sample were compared to those of 1685 controls. RESULTS: Thirty percent of BCSs reported insomnia symptoms in 2004, and arm/shoulder problems were significantly associated with insomnia, as were established variables in bivariate analyses. In 2004 only regular use of hypnotics remained associated with insomnia in multivariate analysis. In bivariate analysis arm/shoulder pain and restricted mobility in 2004 were significant predictors of insomnia in 2007. Only insomnia in 2004 was a significant predictor in multivariate analysis. In bivariate analyses BCSs and controls had several common factors associated with insomnia, but only regular use of hypnotics was common in multivariate analysis. CONCLUSIONS: Arm/shoulder problems are factors to consider in BCSs with insomnia, particularly arm/shoulder pain. Factors associated with insomnia in BCSs and general population controls are partially overlapping. PMID- 21645873 TI - The impact of atypical antipsychotic use on obstructive sleep apnea: a pilot study and literature review. AB - BACKGROUND: Limited evidence links atypical antipsychotics (AAs) use to sleep related respiratory dysfunction and greater severity of obstructive sleep apnea (OSA). The present paper reviews the published evidence and examines the impact of AA use on the presence and severity of OSA among subjects with clinically suspected OSA after adjusting for several confounds. METHODS: Archives of the University of Iowa Sleep Laboratory from 2005 to 2009 were searched for patients using AAs at the time of diagnostic polysomnogram (PSG). PSG data of the 84 AA users with heterogeneous psychiatric disorders (of these 20 diagnosed only with depression) were subsequently compared to PSG data of two randomly selected, non AA user groups from the same patient pool: (i) 200 subjects with a depressive disorder as the only psychiatric diagnosis, and (ii) 331 mentally healthy controls. PSG data were analyzed adjusting for known demographic, medical, and psychiatric risk factors for OSA. RESULTS: Prevalence and severity of OSA did not differ significantly across three groups. Sex, age, body mass index (BMI), and neck circumference (NC) independently predicted OSA. Odds ratio for OSA in the subset of AA users carrying the diagnosis of depression (n=20) compared with subjects without mental illness was 4.53 (p<.05). By contrast, AA users without depression or those with multiple psychiatric diagnoses including depression did not show a statistically significantly elevated OSA risk. CONCLUSIONS: AA use in subjects with depression appears to increase the risk of OSA after controlling for known predisposing factors. PMID- 21645874 TI - Sleep quality and characteristics of college students who use prescription psychostimulants nonmedically. AB - OBJECTIVE: Although psychostimulants have been shown to affect sleep in experimental studies, the relation between nonmedical psychostimulant use and sleep has not been examined. Our goal was to describe the sleep quality and characteristics of college students who use psychostimulant medications nonmedically. METHODS: We surveyed 492 college students about their sleep quality and psychostimulant use characteristics. Sleep quality was assessed using the Pittsburgh Sleep Quality Index (PSQI); psychostimulant use and characteristics were measured via a survey developed for this study. RESULTS: College students who self-reported current or a history of nonmedical psychostimulant use reported worse subjective sleep quality, sleep disturbance, and global PSQI scores than nonusers. The most commonly reported reason for nonmedical psychostimulant use was to improve work performance and concentration. CONCLUSIONS: These data demonstrate worse sleep quality among nonmedical psychostimulant users. The likely consequence of increased daytime sleepiness helps to confirm the known public health concern of nonmedical prescription psychostimulant use among college students. PMID- 21645875 TI - Interaction between stimulus-driven orienting and top-down modulation in attentional capture. AB - The issue whether attentional capture is determined by top-down factors or can be stimulus-driven remains unsolved. To examine this, we used a spatial cueing paradigm with set size four and eight in which a color target is preceded by an uninformative cue (color or onset) that either matches or does not match the target-defining feature. The critical manipulation lies in adding no-cue trials to make the subsequent first with-cue trial unexpected, which reveals the stimulus-driven component. For the onset cue, the first-trial analysis indicated attentional capture at set size four and eight, whereas results from the average data indicated attentional capture at set size eight but not at set size four. For the color cue, attentional capture was found in the average data but not in the first trial, regardless of set size. These results can be explained by the interactive processing of early stimulus-driven activation followed by top-down modulation. PMID- 21645876 TI - Aggregation in colloidal suspensions: effect of colloidal forces and hydrodynamic interactions. AB - The forces acting in colloidal suspensions and affecting their stability and aggregation kinetics are considered. The approximations used for these forces in numerical simulations and the importance of the balanced account for both colloidal forces and hydrodynamic interactions are discussed. As an example the results of direct numerical simulations of kinetics of aggregation either with account for hydrodynamic interaction between particles or without it are compared by varying the parameters of the interaction potential between particles and fraction of solid. Simulations are based on the Langevin equations with pairwise interaction between particles and take into account Brownian, hydrodynamic and colloidal forces. It is confirmed that the neglecting of hydrodynamic interaction results in an accelerated growth of aggregates. The results of numerical simulations of aggregation kinetics are compared with well known analytical solutions. PMID- 21645878 TI - Increases in extracellular serotonin and dopamine metabolite levels in the basal forebrain during sleep deprivation. AB - The basal forebrain (BF) is an important mediator of cortical arousal, which is innervated by all ascending arousal systems. During sleep deprivation (SD) a site specific accumulation of sleep factors in the BF results in increased sleep pressure (Kalinchuk et al., 2006; Porkka-Heiskanen et al., 1997; Porkka-Heiskanen et al., 2000). However, animals are able to stay awake and even increase their neuronal activity in the BF and cortex during SD, suggesting increased activity of the ascending arousal systems to counteract the effect of sleep pressure. This study used in vivo microdialysis to measure the effect of a 6h SD, by "gentle handling" in freely moving rats, on the extracellular levels of serotonin and dopamine metabolites (5-HIAA, and DOPAC and HVA respectively) in the BF. Additionally, because glucocorticoids can interact with monoaminergic neurotransmission, and SD could be stressful, corticosterone levels were measured. We found an increase in extracellular serotonin and dopamine metabolite levels (n=8, p<=0.05). No interaction between corticosterone and the monoaminergic systems was apparent. Extracellular corticosterone levels showed no increase during the first 3h of SD, and the subsequent increase (n=8, p<=0.05) did not result in values exceeding the normal diurnal maximum, indicating that no substantial stress was induced. The results demonstrate that SD increases extracellular dopamine and serotonin metabolites in the BF, suggesting increased activity of the ascending arousal systems. It remains to be investigated what the specific roles of the dopaminergic and serotonergic ascending arousal systems are in BF-mediated cortical arousal. PMID- 21645877 TI - Modulation of dendritic spines and synaptic function by Rac1: a possible link to Fragile X syndrome pathology. AB - Rac1, a protein of the Rho GTPase subfamily, has been implicated in neuronal and spine development as well as the formation of synapses with appropriate partners. Dendrite and spine abnormalities have been implicated in several psychiatric disorders such as Fragile X syndrome, where neurons show a high density of long, thin, and immature dendritic spines. Although abnormalities in dendrites and spines have been correlated with impaired cognitive abilities in mental retardation, the causes of these malformations are not yet well understood. Fragile X syndrome is the most common type of inherited mental retardation caused by the absence of FMRP protein, a RNA-binding protein implicated in the regulation of mRNA translation and transport, leading to protein synthesis. We suggest that FMRP might act as a negative regulator on the synthesis of Rac1. Maintaining an optimal level of Rac1 and facilitating the reorganization of the cytoskeleton likely leads to normal neuronal morphology during activity-dependent plasticity. In our study, we first demonstrated that Rac1 is not only associated but necessary for normal spine development and long-term synaptic plasticity. We further showed that, in Fmr1 knockout mice, lack of FMRP induces an overactivation of Rac1 in the mouse brain and other organs that have been shown to be altered in Fragile X syndrome. In those animals, pharmacological manipulation of Rac1 partially reverses their altered long-term plasticity. Thus, regulation of Rac1 may provide a functional link among deficient neuronal morphology, aberrant synaptic plasticity and cognition impairment in Fragile X syndrome. PMID- 21645879 TI - Individual differences in working memory capacity determine the effects of oculomotor task load on concurrent word recall performance. AB - In this study, the interaction between individual differences in working memory capacity, which were assessed by the Korean version of the California Verbal Learning Test (K-CVLT), and the effects of oculomotor task load on word recall performance are examined in a dual-task experiment. We hypothesized that varying levels of oculomotor task load should result in different demands on cognitive resources. The verbal working memory task used in this study involved a brief exposure to seven words to be remembered, followed by a 30-second delay during which the subject carried out an oculomotor task. Then, memory performance was assessed by having the subjects recall as many words as possible. Forty healthy normal subjects with no vision-related problems carried out four separate dual tasks over four consecutive days of participation, wherein word recall performances were tested under unpredictable random SPEM (smooth pursuit eye movement), predictive SPEM, fixation, and eyes-closed conditions. The word recall performance of subjects with low K-CVLT scores was significantly enhanced under predictive SPEM conditions as opposed to the fixation and eyes-closed conditions, but performance was reduced under the random SPEM condition, thus reflecting an inverted-U relationship between the oculomotor task load and word recall performance. Subjects with high K-CVLT scores evidenced steady word recall performances, regardless of the type of oculomotor task performed. The concurrent oculomotor performance measured by velocity error did not differ significantly among the K-CVLT groups. However, the high-scoring subjects evidenced smaller phase errors under predictive SPEM conditions than did the low-scoring subjects; this suggests that different resource allocation strategies may be adopted, depending on individuals' working memory capacity. PMID- 21645880 TI - Neural correlates of evaluating hazards of high risk. AB - In personal and in society related context, people often evaluate the risk of environmental and technological hazards. Previous research addressing neuroscience of risk evaluation assessed particularly the direct personal risk of presented stimuli, which may have comprised for instance aspects of fear. Further, risk evaluation primarily was compared to tasks of other cognitive domains serving as control conditions, thus revealing general risk related brain activity, but not such specifically associated with estimating a higher level of risk. We here investigated the neural basis on which lay-persons individually evaluated the risk of different potential hazards for the society. Twenty healthy subjects underwent functional magnetic resonance imaging while evaluating the risk of fifty more or less risky conditions presented as written terms. Brain activations during the individual estimations of 'high' against 'low' risk, and of negative versus neutral and positive emotional valences were analyzed. Estimating hazards to be of high risk was associated with activation in medial thalamus, anterior insula, caudate nucleus, cingulate cortex and further prefrontal and temporo-occipital areas. These areas were not involved according to an analysis of the emotion ratings. In conclusion, we emphasize a contribution of the mentioned brain areas involved to signal high risk, here not primarily associated with the emotional valence of the risk items. These areas have earlier been reported to be associated with, beside emotional, viscerosensitive and implicit processing. This leads to assumptions of an intuitive contribution, or a "gut-feeling", not necessarily dependent of the subjective emotional valence, when estimating a high risk of environmental hazards. PMID- 21645881 TI - Bi- and trivalent glycopeptide mannopyranosides as inhibitors of type 1 fimbriae mediated bacterial adhesion: variation of valency, aglycon and scaffolding. AB - In order to test relevant structural parameters for effective inhibition of mannose-specific bacterial adhesion, bi- and trivalent glycopeptide alpha-D mannopyranosides were synthesized that differ in their conformational properties as well as in the spatial arrangement of attached mannosyl residues. They were tested in an inhibition adhesion assay with fluorescent Escherichia coli bacteria and testing results were referenced to the inhibitory potency of methyl alpha-D mannopyranoside. It was shown, that besides the nature of the mannoside aglycon moiety, scaffolding of alpha-D-mannopyranosides on a peptide backbone was important for the performance of the synthesized glycopeptides as inhibitors of bacterial adhesion. PMID- 21645882 TI - Trans-sialidase and mucins of Trypanosoma cruzi: an important interplay for the parasite. AB - A dense glycocalix covers the surface of Trypanosoma cruzi, the agent of Chagas disease. Sialic acid in the surface of the parasite plays an important role in the infectious process, however, T. cruzi is unable to synthesize sialic acid or the usual donor CMP-sialic acid. Instead, T. cruzi expresses a unique enzyme, the trans-sialidase (TcTS) involved in the transfer of sialic acid from host glycoconjugates to mucins of the parasite. The mucins are the major glycoproteins in the insect stage epimastigotes and in the infective trypomastigotes. Both, the mucins and the TcTS are anchored to the plasma membrane by a glycosylphosphatidylinositol anchor. Thus, TcTS may be shed into the bloodstream of the mammal host by the action of a parasite phosphatidylinositol-phospholipase C, affecting the immune system. The composition and structure of the sugars in the parasite mucins is characteristic of each differentiation stage, also, interstrain variations were described for epimastigote mucins. This review focus on the characteristics of the interplay between the trans-sialidase and the mucins of T. cruzi and summarizes the known carbohydrate structures of the mucins. PMID- 21645883 TI - Enzymatic synthesis and properties of uridine-5'-O-(2-thiodiphospho)-N acetylglucosamine. AB - This paper describes an enzymatic approach to obtain a thio-containing UDP-GlcNAc analog. We use an assay based on capture of the carbohydrate and analysis by mass spectrometry to quantitatively characterize the activity of this unnatural sugar donor in a LgtA-mediated glycosylation reaction. PMID- 21645884 TI - Synthesis and characterization of poly-O-methyl-[n]-polyurethane from a d glucamine-based monomer. AB - Aminoalditol 1-amino-1-deoxy-D-sorbitol (1) was readily converted into 2,3,4,5 tetra-O-methyl derivative 5, a key precursor of a sugar-based [n]-polyurethane. For the polymerization, the free amino or primary hydroxyl groups of 5 were selectively activated and employed as starting monomers in two alternative procedures. Thus, the amino function of 5 was converted into the isocyanate derivative by treatment with di-tert-butyltricarbonate, and polymerized in situ in the presence of Zr(IV) acetylacetonate. The resulting poly(1-amino-1-deoxy 2,3,4,5-tetra-O-methyl-D-sorbitol)urethane (8) had a moderate molecular weight and showed the presence of urea units. The alternative synthesis of 8 involved the activation of the free hydroxyl group of 5 as the corresponding phenylcarbonate. The polymerization of this alpha-amino-omega-phenylcarbonate alditol monomer does not require a metal catalyst. The resulting material exhibited an improved molecular weight and higher purity than that obtained via the isocyanate. [n]-polyurethane 8 was highly soluble in water as well as in common organic solvents (chloroform, acetone, ethyl acetate, etc) and was obtained as an amorphous material which was characterized thermally and spectroscopically. PMID- 21645885 TI - 1-Deoxy-D-galactonojirimycins with dansyl capped N-substituents as beta galactosidase inhibitors and potential probes for GM1 gangliosidosis affected cell lines. AB - Two simple and reliably accessible intermediates, N-carboxypentyl- and N aminohexyl-1-deoxy-D-galactonojirimycin were employed for the synthesis of a set of terminally N-dansyl substituted derivatives. Reaction of the terminal carboxylic acid of N-carboxypentyl-1-deoxy-D-galactonojirimycin with N-dansyl-1,6 diaminohexane provided the chain-extended fluorescent derivative. Employing bis(6 dansylaminohexyl)amine, the corresponding branched di-N-dansyl compound was obtained. Partially protected N-aminohexyl-1-deoxy-D-galactonojirimycin served as intermediate for two additional chain-extended fluorescent 1-deoxy-D galactonojirimycin (1-DGJ) derivatives featuring terminal dansyl groups in the N alkyl substituent. These new compounds are strong inhibitors of d-galactosidases and may serve as leads en route to pharmacological chaperones for GM1 gangliosidosis. PMID- 21645886 TI - Experimental observations on the regioselectivity of glycosylation of a 4,6-diol system in the beta-D-mannopyranosyl unit of a N-glycan pentasaccharide core structure. AB - The regioselectivity of glycosylation of a 4,6-diol system in the beta mannopyranosyl unit of a N-glycan pentasaccharide core structure is found to be strongly dependent on the structure of the glycosyl donor. While glycosylation with a 2-O-acetyl-D-mannopyranosyl trichloroacetimidate and with a d mannopyranosyl (alpha1->3) 2-O-acetyl mannopyranosyl trichoroacetimidate regioselectively occurs at the primary OH-6 position, reaction with d mannopyranosyl (alpha1->6) mannopyranosyl 2-O-benzoyl, 2-O-acetyl and 2-O pivaloyl trichloroacetimidate results in approximately 1:1 mixture of regioisomers at primary OH-6 and secondary OH-4 positions. PMID- 21645887 TI - Influence of the solvent in low temperature glycosylations with O-(2,3,5,6-tetra O-benzyl-beta-D-galactofuranosyl) trichloroacetimidate for 1,2-cis alpha-D galactofuranosylation. AB - Glycosylation studies for the construction of 1,2-cis alpha-linkages with O (2,3,5,6-tetra-O-benzyl-beta-D-galactofuranosyl) trichloroacetimidate (1) and several acceptors, including D-mannosyl and l-rhamnosyl derivatives were performed. The reactions were conducted at low temperatures using CH(2)Cl(2), Et(2)O, and acetonitrile as solvents. A non-participating solvent such as CH(2)Cl(2) at -78 degrees C, favored the alpha-D-configuration. In contrast, acetonitrile strongly favored the beta-D-configuration, whereas no selectivities were observed with Et(2)O. The use of thiophene as an additive did not enhance the alpha-D-selectivity as in the pyranose counterpart. Although selectivities strongly depended on the acceptor, trichloroacetimidate 1 constitutes a valuable donor for the synthesis of alpha-D-Galf-(1->2)-l-Rha and alpha-D-Galf-(1->6)-D Man. As these motifs are present in pathogenic microorganisms, these procedures described here are useful for the straightforward synthesis of natural oligosaccharides. PMID- 21645888 TI - Sodium halide complexes of ribose derivatives and their unusual crystal structures. AB - Crystalline complexes of D-ribose, D-ribono-1,4-lactone and methyl beta-D ribopyranoside with sodium halides were synthesized and some of their crystal structures determined. Crystal structures of two lactone complexes and a methyl beta-D-ribopyranoside reveal the mode of the salt binding and the intricate interplay of cation coordination and hydrogen bonding in these complexes. When complexed with NaBr, the ribopyranoside is in the (1)C(4) shape whereas ribose with no salt present has the (4)C(1) shape. It is also demonstrated that such complexes can be easily prepared in solid state reaction using a ball mill. PMID- 21645890 TI - Optimal timing of oocyte maturation and its relationship with the spindle assembly and developmental competence of in vitro matured human oocytes. AB - OBJECTIVE: To develop and test a simple, feasible approach to improve spindle assembly and developmental competence of human in vitro matured oocytes by parthenogenetic activation in maturation medium. DESIGN: Prospective, randomized study. SETTING: University research laboratory-based assisted reproductive technology laboratory. PATIENT(S): Four hundred thirty-two patients with male factor infertility undergoing intracytoplasmic sperm injection (ICSI) cycles. INTERVENTION(S): In vivo failed-to-mature oocytes from the ICSI cycles were divided into four groups according to differing duration after the extrusion of first polar body (0-1, 2-3, 4-5, and 8-9 hours). Oocytes spindles in each group were immunostained for alpha-tubulin and chromosomes, and observed by confocal microscopy. MAIN OUTCOME MEASURE(S): Rate from pronuclear stage to blastocyst, embryo grading at the eight-cell and blastocyst stages, and spindle assembly. RESULT(S): There was a statistically significantly higher rate of development at the eight-cell and blastocyst stages in the 2- to 3-hour and 4- to 5-hour groups. The grading results at the eight-cell and blastocyst stages also showed that the proportion of embryos of high quality was similar among the 2- to 3-hour and 4- to 5-hour groups, and in these groups it was statistically significantly higher than the 0- to 1-hour and 8- to 9-hour groups. The results of immunofluorescence demonstrated that there was a statistically significantly higher rate of normal spindle assembly in the 2- to 3-hour and 4- to 5-hour groups. CONCLUSION(S): Optimal timing of maturation benefits the development of competence of in vitro matured oocytes by promoting normal spindle assembly. PMID- 21645891 TI - Reduced neurotrophin receptor tropomyosin-related kinase A expression in human granulosa cells: a novel marker of diminishing ovarian reserve. AB - OBJECTIVE: To determine whether human mural and cumulus granulosa cell neurotrophin and neurotrophin receptor content correlate to ovarian reserve markers. DESIGN: Prospective, laboratory-based study. SETTING: Academic assisted reproductive technology (ART) program. PATIENT(S): Twenty-three women undergoing ART. INTERVENTION(S): Mural and cumulus granulosa cells were collected from women undergoing oocyte retrieval during ART cycles. Relative messenger RNA (mRNA) levels of neurotrophins and their receptors were measured by quantitative reverse transcription-polymerase chain reaction and correlated to serum antimullerian hormone (AMH) levels and the number of oocytes retrieved. MAIN OUTCOME MEASURE(S): Number of oocytes retrieved. RESULT(S): Mural and cumulus granulosa cell nerve growth factor receptor tropomyosin-related kinase A (TrkA) mRNA correlated strongly to the number of oocytes retrieved. Similarly, higher serum AMH was associated with higher cumulus granulosa cell TrkA mRNA. Both mural and cumulus granulosa cell p75(NTR)/TrkA ratios were lower in women with higher serum AMH, and the number of oocytes retrieved was greater among women with low p75(NTR)/TrkA ratio. No significant associations were found between brain-derived neurotrophic factor (BDNF) and its specific receptor tropomyosin-related kinase B (TrkB) and ovarian reserve markers. Although BDNF and TrkB expression were higher in cumulus compared with mural granulosa cells, no such association was found between TrkA and granulosa cells. Antimullerian hormone and cumulus TrkA mRNA, in a model incorporating both, correlated strongly to the number of oocytes retrieved (R(2) = 0.84). CONCLUSION(S): Cumulus TrkA and p75(NTR) mRNA correlate to ovarian reserve, whereas BDNF and TrkB are associated with the type of granulosa cell. PMID- 21645892 TI - Does meiotic spindle normality predict improved blastocyst development, implantation and live birth rates? AB - OBJECTIVE: To investigate the relationship between spindle normality and: 1) blastocyst formation; 2) implantation rates; and 3) live birth rates. DESIGN: Prospective observational study. SETTING: A private fertility clinic. PATIENT(S): One hundred patients <=40 years old undergoing intracytoplasmic sperm injection (ICSI). INTERVENTION(S): Meiotic spindles were imaged before ICSI with the use of Oosight microscopy. With the use of specific criteria, spindle normality was independently assessed by two researchers. Blastocysts were chosen for transfer by standard light microscopic morphologic criteria by researchers who were blinded to the spindle assessment. MAIN OUTCOME MEASURE(S): 1) Blastocyst formation; 2) implantation; and 3) live birth rates. RESULT(S): A total of 808/920 oocytes were metaphase II. Of those, 711 (88%) had a visible spindle: 205 (29%) were normal spindles (NS) and 506 (71%) abnormal spindles (AS). Fertilization rates were significantly higher in NS oocytes. Although NS and AS oocytes both formed morphologically good-quality blastocysts, implantation and live birth rates were higher from NS oocytes. All ongoing pregnancies resulted from NS oocytes. CONCLUSION(S): Spindle assessment with polarized light microscopy provides an early predictor of the pregnancy potential of that oocyte. AS oocytes can form good-quality blastocysts, but these appear to have little chance of live birth. Therefore, spindle assessment should improve the selection of the best embryo for single embryo transfer. PMID- 21645893 TI - Primate preimplantation embryo is a target for relaxin during early pregnancy. AB - OBJECTIVE: To determine whether preimplantation embryos are targets for relaxin secreted from the corpus luteum of the menstrual cycle. DESIGN: Rhesus monkey oocytes obtained from females undergoing controlled ovarian hyperstimulation were inseminated, and the resulting embryos were cultured in medium with or without recombinant human relaxin (20 ng/mL) for 8 days. SETTING: Research laboratory. ANIMAL(S): Rhesus monkey. INTERVENTION(S): Controlled ovarian stimulation to obtain oocytes for in vitro-produced embryos that were cultured with or without human recombinant relaxin. MAIN OUTCOME MEASURE(S): Rate of blastocyst development, percentage of blastocysts, and inner cell mass/trophectoderm cell ratio were measured on day 8 of culture. The presence of relaxin receptor (RXFP1) messenger RNA in eight-cell embryos was observed by array hybridization. RESULT(S): RXFP1 receptor expression was localized to the inner cell mass of blastocysts, as shown by immunohistochemistry. The percentage of embryos that developed to blastocyst and the inner cell mass/trophectoderm cell ratio was unchanged with relaxin supplementation; however, the relaxin-treated embryos developed into blastocysts significantly sooner than untreated embryos. CONCLUSION(S): These results are the first evidence that the preimplantation primate embryo is a target for relaxin and that the addition of relaxin to in vitro culture medium enhances rhesus monkey embryo development. PMID- 21645894 TI - Increasing burden of institutional review in multicenter clinical trials of infertility: the Reproductive Medicine Network experience with the Pregnancy in Polycystic Ovary Syndrome (PPCOS) I and II studies. AB - Many clinical investigators think that the burden of Institutional Review Board (IRB) requirements has been consistently increasing over recent years, although there are few objective data describing these trends. Over a period of 7 years, the Reproductive Medicine Network observed a significant increase in the size and requirements of IRB submissions and significant variability of IRB performance in reviewing multicenter trials. These additional regulatory and administrative demands represent substantial burdens to researchers and to the IRBs themselves. It is timely to consider whether these changes better protect the interests and safety of human research participants. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov NCT00068861 and NCT00719186. PMID- 21645895 TI - Plasma leptin and adiponectin levels in hormone replacement therapy and contraception: effects of different progestogens. AB - OBJECTIVE: To evaluate the impact of hormone replacement therapy and oral contraceptives containing different progestogens on adiponectin and leptin serum levels. DESIGN: Prospective, longitudinal, semi-randomized study. SETTING: General gynecology clinic of a university hospital. PATIENT(S): Forty-five healthy postmenopausal and 45 healthy premenopausal women. INTERVENTION(S): Thirty postmenopausal women were randomized to receive either drospirenone (DRSP) 2 mg + E(2) 1 mg (n = 15) or norethisterone acetate (NETA) 0.5 mg + E(2) 1 mg (n = 15). Thirty premenopausal women were randomized to receive either ethinilestradiol (EE) 0.020 mg + DRSP 3 mg (n = 15) or EE 0.020 mg + desogestrel 0.15 mg (n = 15). Furthermore, 15 postmenopausal and 15 premenopausal untreated women served as controls. MAIN OUTCOME MEASURE(S): Adiponectin and leptin serum levels before and after 6 months of hormonal therapy. RESULT(S): No significant differences in leptin levels were detected in any group after 6 months. Adiponectin levels were significantly reduced in the NETA + E(2) group and increased in the EE + DRSP group, while remaining unmodified in all other groups. CONCLUSION(S): Hormone replacement therapy with NETA, but not with DRSP, decreases serum adiponectin levels. Oral contraceptives containing DRSP increase serum adiponectin levels. PMID- 21645896 TI - Inter- and intraobserver agreement on the Load Sharing Classification of thoracolumbar spine fractures. AB - The Load Sharing Classification (LSC) allocates one to three points to each of three different radiological characteristics of traumatic thoracolumbar fractures: the vertebral body involved in the fracture, the displacement of the fracture parts and the kyphotic deformity. Added up, a minimal score of three and a maximal score of nine can be obtained. When the LSC score is three to six, a short segment pedicle screw fixation suffices. When the LSC score is seven to nine, a high rate of failure in patients with a short segment pedicle screw fixation exists. In these cases an anterior stabilising procedure of the spine is advised. The LSC has been examined by Dai and Jin, who claim an almost perfect inter- and intraobserver agreement, according to the Landis and Koch criteria. Dai and Jin only present results for the separate three items of the LSC and for the total LSC scores. Observer agreement for the two LSC score categories (three to six and seven to nine) have not been studied. The aim of this study is to study the inter- and intraobserver agreement of the LSC for the total score, the three separate items and also for the two LSC score categories. Three observers determine twice the LSC scores of forty traumatic thoracolumbar fractures. The average standard Cohen's kappa values for the separate LSC items range between 0.06 and 0.48. For the total LSC score the average standard Cohen's kappa and weighted kappa values are 0.22 and 0.67 respectively. For the two LSC score categories, there is unanimous agreement in 55% of the cases and a majority agreement in 40%. In the remaining 5% of the fractures there is a split decision. Standard Cohen's kappa value for the two LSC score categories is 0.53. The standard Cohen's kappa values can be rated as fair to moderate. From these data it can be concluded that the inter- and intraobserver reliability of the Load Sharing Classification of Spinal fractures can be rated as fair. PMID- 21645897 TI - Validation and clinical impact of paediatric pressure ulcer risk assessment scales: A systematic review. AB - BACKGROUND: Pressure ulcer risk assessment using an age-appropriate, valid and reliable tool is recommended for clinical paediatric practice. OBJECTIVES: (1) What PU risk scales for children currently exist? (2) What is the diagnostic accuracy of their scores? (3) Are the scores reliable and what is the degree of agreement? (4) What is the clinical impact of risk scale scores in paediatric practice? DESIGN: Systematic review. DATA SOURCES: MEDLINE (1950 to December 2010), EMBASE (1989 to December 2010), CINAHL (1982 to December 2010), reference lists. REVIEW METHODS: Two reviewers independently screened databases, selected and evaluated articles and studies. Diagnostic accuracy, reliability/agreement, and experimental studies investigating the performance and clinical impact of PU risk scale scores in the paediatric population (0-18 years) were included. PU development was used as reference standard for diagnostic accuracy studies. Methodological quality of the validity and reliability studies was assessed based on the QUADAS and QAREL checklists. RESULTS: The search yielded 1141 hints. Finally, 15 publications describing or applying 12 paediatric pressure ulcer risk scales were included. Three of these scales (Neonatal Skin Risk Assessment Scale for Predicting Skin Breakdown, Braden Q Scale, Burn Pressure Skin Risk Assessment Scale) were investigated in prospective validation studies. Empirical evidence about interrater reliability and agreement is available for four instruments (Neonatal Skin Risk Assessment Scale for Predicting Skin Breakdown, Starkid Skin Scale, Glamorgan Scale, Burn Pressure Ulcer Risk Assessment Scale). No studies were identified investigating the clinical impact. CONCLUSIONS: Sound empirical evidence about the performance of paediatric pressure ulcer risk assessment scales is lacking. Based on the few results of this review no instrument can be regarded as superior to the others. Whether the application of pressure ulcer risk assessment scales reduces the pressure ulcer incidence in paediatric practice is unknown. Maybe clinical judgement is more efficient in evaluating pressure ulcer risk than the application of risk scale scores. PMID- 21645898 TI - Identification of a gene variant in the master regulator of lipid metabolism SREBP-1 in a family with a novel form of severe combined hypolipidemia. AB - OBJECTIVE: Alterations of lipid metabolism play a pivotal role in the development of atherosclerosis and its complications, today's major mortality risks. The predominant regulators controlling cholesterol- and fatty acids synthesis in liver are the sterol regulatory element-binding proteins (SREBPs), a family of transcription factors that were formerly identified as cholesterol sensor for LDLR gene expression. Variation of gene structure in these genes might therefore indicate a predisposition to develop complications like myocardial infarction and stroke. METHODS: We investigated 190 unrelated German subjects, including 69 subjects with LDL-cholesterol <55mg/dl, for mutations in SREBP genes SREBF-1 and SREBF-2 by direct sequencing. The impact on SREBP functionality was analyzed by protein biochemical analyses, promoter reporter gene assays and gene expression studies. RESULTS: A missense mutation in SREBF-1 (c.332 C>T; P111L) was identified in a subject with LDL-cholesterol <5mg/dl. Examination of the subject's family confirmed the mutation in two of three siblings. Detailed clinical evaluation of these subjects disclose a novel form of primary combined hypolipidemia only in SREBP-1a P111L carriers, characterized by low levels of apoB and apoA1, low triglyceride, LDL-cholesterol and HDL-cholesterol levels. Functional analyses indicated that the mutation abolishes phosphorylation of SREBP-1. As a consequence transcriptional activation of classical target genes, i.e. LDLR, HMG-CoAR, FAS, ABCA1, but also MTTP, was dramatically reduced. CONCLUSIONS: Phosphorylation of SREBP-1, the master regulator of genes for central rate limiting enzymes of cholesterol and lipid metabolism, appears to be a biological principle with clinical implications. PMID- 21645899 TI - Large-artery stiffness: a reversible marker of cardiovascular risk in primary hyperparathyroidism. AB - OBJECTIVE: Patients with primary hyperparathyroidism (pHPT) are at increased risk of cardiovascular mortality. We investigated whether aortic stiffness, an early marker of arteriosclerosis and a strong predictor of cardiovascular risk, is increased in pHPT, and whether it improves after parathyroidectomy. METHODS: Twenty-four patients with mild pHPT (age 56 +/- 10 years, blood pressure 136/85 mmHg, serum calcium 2.55-3.00 mmol/L) and 48 control subjects individually matched with cases by age, sex and blood pressure underwent aortic (carotid femoral) and upper-limb (carotid-radial) pulse wave velocity (PWV) determination by applanation tonometry in a case-control study. Subjects with renal disease, diabetes, treated hypertension or overt cardiovascular disease were excluded from the study. Seventeen of the patients with pHPT were re-examined 4 weeks after surgical parathyroidectomy. RESULTS: Aortic PWV was significantly higher among pHTP patients (11.4 +/- 2 vs 9.6 +/- 2 m/s, p<0.001). In a conditional logistic regression analysis, pHPT was independently associated with an increased risk of having an aortic PWV >12 m/s (odds ratio 3.28, 95% confidence interval 1.21 8.93). As expected, surgery was accompanied by a reduction in serum calcium (from 2.77 +/- 0.2 to 2.25 +/- 0.1 mmol/L, p<0.001) and parathyroid hormone (from 29.6 +/- 10 to 3.3 +/- 2 pmol/L, p<0.001). Aortic PWV decreased after surgery (from 10.9 +/- 2 to 9.8 +/- 2 m/s, p=0.003). The change in aortic PWV remained significant also after adjustment for changes in blood pressure (p<0.01). Changes in upper-limb PWV generally paralleled those in aortic PWV. CONCLUSION: pHPT is associated with increased aortic stiffness, which improves after parathyroidectomy. Our data demonstrate that aortic stiffness may improve upon removal of hyperparathyroid stimuli. PMID- 21645900 TI - Enantioselective analysis of ibuprofen, ketoprofen and naproxen in wastewater and environmental water samples. AB - A highly sensitive and reliable method for the enantioselective analysis of ibuprofen, ketoprofen and naproxen in wastewater and environmental water samples has been developed. These three pharmaceuticals are chiral molecules and the variable presence of their individual (R)- and (S)-enantiomers is of increasing interest for environmental analysis. An indirect method for enantioseparation was achieved by the derivatization of the (R)- and (S)-enantiomers to amide diastereomers using (R)-1-phenylethylamine ((R)-1-PEA). After initial solid phase extraction from aqueous samples, derivatization was undertaken at room temperature in less than 5 min. Optimum recovery and clean-up of the amide diastereomers from the derivatization solution was achieved by a second solid phase extraction step. Separation and detection of the individual diastereomers was undertaken by gas chromatography-tandem mass spectrometry (GC-MS/MS). Excellent analyte separation and peak shapes were achieved for the derivatized (R)- and (S)-enantiomers for all three pharmaceuticals with peak resolution, R(s) is in the range of 2.87-4.02 for all diastereomer pairs. Furthermore, the calibration curves developed for the (S)-enantiomers revealed excellent linearity (r(2) >= 0.99) for all three compounds. Method detection limits were shown to be within the range of 0.2-3.3 ng L(-1) for individual enantiomers in ultrapure water, drinking water, surface water and a synthetic wastewater. Finally, the method was shown to perform well on a real tertiary treated wastewater sample, revealing measurable concentrations of both (R)- and (S)-enantiomers of ibuprofen, naproxen and ketoprofen. Isotope dilution using racemic D(3) ibuprofen, racemic D(3)-ketoprofen and racemic D(3)-naproxen was shown to be an essential aspect of this method for accurate quantification and enantiomeric fraction (EF) determination. This approach produced excellent reproducibility for EF determination of triplicate tertiary treated wastewater samples. PMID- 21645901 TI - Assessment of the degradation of polyurethane foams after artificial and natural ageing by using pyrolysis-gas chromatography/mass spectrometry and headspace solid phase microextraction-gas chromatography/mass spectrometry. AB - Polyurethane foams are widely present in museum collections either as part of the artefacts, or as a material for their conservation. Unfortunately many of PU foam artefacts are in poor condition and often exhibit specific conservation issues. Their fast thermal and photochemical degradations have been the aim of previous researches. It is now accepted that hydrolysis predominates for polyester-based polyurethane PU(ES) whereas oxidation is the principal cause of degradation for polyether-based polyurethane PU(ET) variety. Only a few studies have been devoted to volatile organic compounds (VOCs) emitted by polyurethanes and, to our knowledge, none were performed on polyurethane foams by using headspace-solid phase microextraction (HS-SPME). The objective of the work described here is to assess the impact of some environmental factors (humidity, temperature and daylight) on the degradation of PU foams by evaluating their volatile fractions. We investigated morphological changes, polymerized fractions and volatile fractions of (i) one modern produced PU(ES) foam and one modern PU(ET) foam artificially aged in different conditions as well as (ii) four naturally aged foams collected from various daily life objects and selected for the representativeness of their analytical data. Characterization procedure used was based on attenuated total reflection Fourier transform infrared spectroscopy (ATR FTIR), pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS) and non-invasive headspace solid-phase microextraction coupled with gas chromatography and mass spectrometry (HS-SPME-GC/MS). In this paper, the formation of alcohol and acid raw products for PU(ES) and glycol derivatives for PU(ET) during natural and artificial ageing is confirmed. These main products can be considered as degradation markers for PU foams. Results show that artificial and natural ageing provide similar analytical results, and confirm that the dominant degradation paths for PU(ES) and for PU(ET) are hydrolysis and photo-oxidation, respectively. Lastly, we highlight that non-invasive HS-SPME-GC/MS analysis allows to distinguish between PU(ES) and PU(ET) at any point of their degradations. PMID- 21645902 TI - Molecularly imprinted polymer with high-fidelity binding sites for the selective extraction of barbiturates from human urine. AB - In this paper we describe the synthesis of a molecularly imprinted polymer (MIP) by precipitation polymerisation, with barbital as the template molecule, and the application of the barbital MIP as a molecularly selective sorbent in the solid phase extraction (SPE) of barbiturates from human urine samples. The MIP was synthesised by precipitation polymerisation using 2,6-bis-acrylamidopyridine as the functional monomer and DVB-80 as the cross-linking agent. The spherical MIP particles produced were 4.2 +/- 0.4 MUm in diameter; a non-imprinted control polymer (NIP) in bead form was 4.8 +/- 0.4 MUm (mean+/-standard deviation) in diameter. The particles were packed into a solid-phase extraction cartridge and employed as a novel sorbent in a molecularly imprinted solid-phase extraction (MISPE) protocol. The MIP showed high selectivity for the template molecule, barbital, a feature which can be ascribed to the high-fidelity binding sites present in the MIP which arose from the use of 2,6-bis-acrylamidopyridine as the functional monomer. However, the MIP also displayed useful cross-selectivity for other barbiturates besides barbital. For real samples, the MIP was applied for the extraction of four barbiturates from human urine. However, due to the high urea concentration in this sample which interfere the proper interaction of barbiturates onto the MIP, a tandem system using a commercially available sorbent was developed. PMID- 21645903 TI - Controllable preparation of high-yield magnetic polymer latex. AB - In order to overcome the low conversion and complex post-treatment, four different polymerization procedures were adopted to prepare the magnetic polymer latexes. The results clearly show that the strategy using magnetic emulsion template-dosage is the most effective and feasible. Based on the optimized procedure, various factors including the type of initiators such as oil soluble initiator, water soluble initiator, redox initiator system, crosslinking agent, functional monomers etc. were systematically studied. Magnetic polymer latex with high monomer conversion of 83% and high magnet content of 31.8% was successfully obtained. Besides, core-shell structured magnetic polymer latex with good film forming property was also prepared, which is promising for potential applications such as magnetic coatings and modification of cementitious materials with controlled polymer location. PMID- 21645904 TI - Selective adsorption of protein molecules on phase-separated sapphire surfaces. AB - Site-selective adsorption of protein molecules was found on sapphire surfaces that exhibit a phase separation into two domains: weakly charged hydrophobic domain and negatively charged hydrophilic one. Ferritin and bovine serum albumin molecules, which are negatively charged in a buffer solution, are adsorbed to the hydrophobic domains. Avidin molecules, which are positively charged, are adsorbed to the other domain. Fibrinogen molecules, which consist of both negative and positive modules, are adsorbed to the whole sapphire surface. Hemoglobin molecules, whose net charge is almost zero, are also adsorbed to the whole surfaces. These results indicate that electrostatic double layer interaction is the primary origin of the observed selectivity. Dependence of protein adsorption or desorption behaviors on the pH value can also be interpreted by the proposed model. PMID- 21645905 TI - Solid-state radical grafting reaction of glycidyl methacrylate and poly(4-methyl 1-pentene) in supercritical carbon dioxide: surface morphology and adhesion. AB - Solid-state radical grafting of glycidyl methacrylate (GMA) onto poly(4-methyl-1 pentene) (PMP) was performed using supercritical carbon dioxide (scCO(2)) impregnation technology. The polymer films were firstly impregnated in the scCO(2) phase with the GMA using benzoyl peroxide as thermal initiator. The grafting degree and surface morphology of the samples may be controlled by the following factors: time, temperature, and pressure of impregnation. A 2(3) factorial design to evaluate the main and interaction effects of such factors on the grafting of the PMP by GMA (grafting response) was elaborated from FTIR data. The superior and inferior limits of the levels were defined on basis of a P-x-y diagram for binary system CO(2)+GMA that provided the location of the transition curves of such a system. Better grafting response was obtained for pressure of 130 bar, temperature of 70 degrees C and time of 7h. The PMP-g-GMA films exhibited a thermal profile similar to that of the unmodified polymer. Adhesion characteristics of polymer films are dependent on grafting degree of GMA. PMID- 21645906 TI - Pointing gestures as a cognitive tool in young children: experimental evidence. AB - This article explores the possible cognitive function associated with pointing gestures from a Vygotskian perspective. In Study 1, 39 children who were 2-4years of age were observed in a solitary condition while solving a mnemonic task with or without an explicit memory demand. A discriminant analysis showed that children used noncommunicative pointing gestures only in the task with an explicit memory demand. In Study 2, 39 children who were 4-6years of age completed an attentional task with and without the possibility of pointing. An analysis of variance showed that only those children who spontaneously pointed to solve the first task performed significantly worse in a second condition where pointing was impossible. These results suggest that besides its social interactive functions, pointing may also subserve private cognitive functions for children. PMID- 21645907 TI - The role of inhibitory control in children's cooperative behaviors during a structured puzzle task. AB - This study examined the role of inhibitory control (measured by Stroop interference) in children's cooperative behaviors during a structured puzzle task. The sample consisted of 250 8-, 10-, and 12-year-olds (117 girls and 133 boys) attending classrooms in three primary schools in Northern Italy. Children individually completed an elaborated Stroop task, were paired with classmates into 125 dyads, and were observed during a 10-min puzzle task. Results confirmed that interaction partners exhibited similar levels of cooperative behaviors, and the cooperative behaviors of children predicted changes in the cooperative behaviors of their partners throughout the puzzle task. Cooperative behaviors of each interaction partner were predicted by the child's own inhibitory control as well as the inhibitory control of the partner. Findings are discussed within a developmental contextual framework. PMID- 21645908 TI - Comparison of biomarkers and parent report of tobacco exposure to predict wheeze. AB - OBJECTIVE: To identify the optimal measure of active and passive prenatal tobacco exposure to predict wheeze in early life. STUDY DESIGN: We conducted a birth cohort study of 398 mother-infant dyads enrolled during the second trimester of pregnancy and followed through age 2 years. We measured tobacco exposure with maternal report, serum cotinine level, and meconium cotinine level. We assessed wheeze with parent report every 6 months. We used a repeated measures logistic regression model. RESULTS: Of 367 children with respiratory data, 26% percent had parent reported active or passive prenatal maternal tobacco exposure, but cotinine was detected in 61% of mothers during pregnancy. Compared with children of mothers in the fifth percentile of tobacco exposure, children of mothers in the 95th percentile had increased odds of wheeze when exposure was measured with maternal serum cotinine level (adjusted OR, 2.6; 95% CI, 1.3-5.2; P < .006) versus meconium cotinine level (adjusted OR, 2.0; 95% CI, 1.0-4.0; P = .04) and total parent-reported exposure (adjusted OR, 1.7; 95% CI, 1.1-2.7; P = .01). CONCLUSIONS: Serum cotinine, a biomarker of tobacco exposure, was more strongly associated with wheeze than parent-reported exposure. Studies that rely on parent report of prenatal tobacco exposure may underestimate risk of wheeze. PMID- 21645909 TI - Use of C-reactive protein in children with newly diagnosed inflammatory bowel disease. AB - C-reactive protein (CRP), a marker for inflammation, was evaluated with other routine blood tests in children with newly diagnosed inflammatory bowel disease. Evaluation of CRP level helped identify additional patients found to have inflammatory bowel disease at endoscopy, although a sizeable number of patients with mild ulcerative colitis had a normal CRP level. PMID- 21645910 TI - Two lumens, one diagnosis. PMID- 21645912 TI - Oxidant/antioxidant balance, minerals status and apoptosis in peripheral blood of dogs naturally infected with Dirofilaria immitis. AB - The present study was aimed to examine lipid peroxides, antioxidants, zinc, copper, iron panels and apoptosis in peripheral blood of dogs with clinical dirofilariosis. In comparison to healthy control, erythrocytic lipid peroxides level and superoxide dismutase activity were significantly (P<0.01) higher in diseased dogs. However, level of reduced glutathione and activities of glutathione peroxidase, glutathione-S-transferase and catalase were significantly (P<0.01) lower in diseased dogs. Blood zinc, copper and iron levels in dogs with dirofilariosis were significantly (P<0.01) lower than the healthy control. The percent apoptotic peripheral leukocytes were significantly higher in dogs with dirofilariosis in comparison with healthy control. From the present study, it is concluded that significant alterations in oxidant/antioxidant balance, mineral status and rate of apoptosis in peripheral blood may be implicated in the pathogenesis of clinical canine dirofilariosis. PMID- 21645911 TI - Screening for sleep-disordered breathing and excessive daytime sleepiness in adolescent girls with polycystic ovarian syndrome. AB - OBJECTIVE: To determine the prevalence and clinical and metabolic correlates of sleep-disordered breathing (SDB) and excessive daytime sleepiness (EDS) in adolescent girls with polycystic ovarian syndrome (PCOS). STUDY DESIGN: Standardized questionnaires were administered to participants with PCOS and age-, sex-, ethnicity-, and body mass index (BMI) z score-matched controls. Medical records were reviewed for anthropometric and metabolic data. RESULTS: We studied 103 participants with PCOS (16.9 +/- 1.5 years) and 90 controls (16.8 +/- 1.7 years). Compared with controls, girls with PCOS had a higher prevalence of SDB (45.6% vs 27.8%; P = .01) and EDS (54.4% vs 35.6%; P < .01). Within PCOS, those with SDB had higher BMI z score (2.1 +/- 0.5 vs 1.7 +/- 0.6; P < .01), higher homeostatic model assessment index (5.1 +/- 2.3 vs 4.1 +/- 3.5; P < .01), and higher prevalence of the metabolic syndrome (MetS) (42.6% vs 16.1%; P = .003) compared with those without SDB. Similarly, participants with PCOS and EDS had a higher BMI z score (2.0 +/- 0.6 vs 1.7 +/- 0.6; P = .03), higher homeostatic model assessment index (5.1 +/- 2.9 vs 3.8 +/- 3.1; P = .01), and higher rate of MetS (39.3% vs. 14.9%; P < .01) compared with those without EDS. MetS was independently associated with SDB (OR, 3.2; 95% CI, 1.0-10.1) and EDS (OR, 4.5; 95% CI, 1.2-16). CONCLUSIONS: SDB and EDS are highly prevalent in adolescent girls with PCOS compared with matched controls. The MetS is independently associated with SDB and EDS in this group. PMID- 21645913 TI - High seroprevalence of Toxoplasma gondii and Neospora caninum in the Common raven (Corvus corax) in the Northeast of Spain. AB - In recent years, multiple cases of aggressive behavior of Common ravens (Corvus corax) have been reported by farmers in Catalonia (NE Spain), including attacking of newborn animals and consumption of dead foetuses. In the present study, seroprevalence of Toxoplasma gondii and Neospora caninum was determined from 113 legally trapped and released Common ravens. T. gondii antibodies were found in 91 (80.5%; CI 95%:72-87) of 113 sera tested by the modified agglutination test. Antibodies to N. caninum were found in 24 (35.8%; IC 95%: 24.5-48.5) of 67 Common ravens tested by an indirect fluorescence antibody test with titers ranging from 1:50 (n=18) to >=1:100 (n=6). To the author's knowledge, this is the first report of the presence of T. gondii and N. caninum antibodies in C. corax. The seroprevalence detected is one of the highest reported worldwide in wild birds, suggesting an important role for this species in the epidemiology of both parasites. PMID- 21645914 TI - Assessment of tumor vasculature for diagnostic and therapeutic applications in a mouse model in vivo using 25-MHz power Doppler imaging. AB - OBJECTIVE: The blood flow rate in the microcirculation associated with angiogenesis plays an important role in the progression and treatment of cancer. Since the microvascular status of tumor vessels can yield useful clinical information, assessing changes in the tumor microcirculation could be particularly helpful for tumor evaluation and treatment planning. METHODS: In this study we used a self-developed 25-MHz ultrasound imaging system with a spatial resolution of 150 MUm for assessing tumor-microcirculation development and the pattern of the vasculature in three tumor-bearing mice in vivo based on power Doppler images. The total Doppler power (DP) and color pixel density (CPD) revealed the presence of functional vessels distributed throughout a tumor volume. The vasculature distributions in the core and periphery were compared to the regulation of vasculature function, which facilitated determination of when the tumor grew rapidly. RESULTS: The data obtained from a quantified analysis of power Doppler images indicated that the tumor vascularity initially increased throughout the tumor. Both DP and CPD increased rapidly in the tumor periphery when the tumor volume exceeded 10mm(3). CONCLUSION: Our preclinical findings suggest that power Doppler imaging could be useful for detecting the changes in tumor vascular perfusion and for determining the optimal treatment timing when a tumor begins its rapid volumetric growth. PMID- 21645915 TI - Mutational analysis of three predicted 5'-proximal stem-loop structures in the genome of tick-borne encephalitis virus indicates different roles in RNA replication and translation. AB - Flavivirus gene expression is modulated by RNA secondary structure elements at the terminal ends of the viral RNA molecule. For tick-borne encephalitis virus (TBEV), four stem-loop (SL) elements have been predicted in the first 180 nucleotides of the viral genome: 5'-SL1, 5'-SL2, 5'-SL3 and 5'-SL4. The last three of these appear to be unique to tick-borne flaviviruses. Here, we report their characterization by mutagenesis in a TBEV luciferase reporter system. By manipulating their thermodynamic properties, we found that an optimal stability of the 5'-SL2 is required for efficient RNA replication. 5'-SL3 formation is also important for viral RNA replication, but although it contains the viral start codon, its formation is dispensable for RNA translation. 5'-SL4 appears to facilitate both RNA translation and replication. Our data suggest that maintenance of the balanced thermodynamic stability of these SL elements is important for temporal regulation of its different functions. PMID- 21645916 TI - Efficiency and energy requirements for the transformation of organic micropollutants by ozone, O3/H2O2 and UV/H2O2. AB - The energy consumptions of conventional ozonation and the AOPs O(3)/H(2)O(2) and UV/H(2)O(2) for transformation of organic micropollutants, namely atrazine (ATR), sulfamethoxazole (SMX) and N-nitrosodimethylamine (NDMA) were compared. Three lake waters and a wastewater were assessed. With p-chlorobenzoic acid (pCBA) as a hydroxyl radical ((*)OH) probe compound, we experimentally determined the rate constants of organic matter of the selected waters for their reaction with (*)OH (k(OH,DOM)), which varied from 2.0 * 10(4) to 3.5 * 10(4) L mgC(-1) s(-1). Based on these data we calculated (*)OH scavenging rates of the various water matrices, which were in the range 6.1-20 * 10(4) s(-1). The varying scavenging rates influenced the required oxidant dose for the same degree of micropollutant transformation. In ozonation, for 90% pCBA transformation in the water with the lowest scavenging rate (lake Zurich water) the required O(3) dose was roughly 2.3 mg/L, and in the water with the highest scavenging rate (Dubendorf wastewater) it was 13.2 mg/L, corresponding to an energy consumption of 0.035 and 0.2 kWh/m(3), respectively. The use of O(3)/H(2)O(2) increased the rate of micropollutant transformation and reduced bromate formation by 70%, but the H(2)O(2) production increased the energy requirements by 20-25%. UV/H(2)O(2) efficiently oxidized all examined micropollutants but energy requirements were substantially higher (For 90% pCBA conversion in lake Zurich water, 0.17-0.75 kWh/m(3) were required, depending on the optical path length). Energy requirements between ozonation and UV/H(2)O(2) were similar only in the case of NDMA, a compound that reacts slowly with ozone and (*)OH but is transformed efficiently by direct photolysis. PMID- 21645917 TI - Adsorption removal of boron in aqueous solutions by amine-modified tannin gel. AB - A tannin gel (TG) synthesized from condensed tannin molecules has a remarkable ability to adsorb various metal ions in aqueous solutions. In the present study, the adsorption removal of boron in solutions at various pHs and temperatures has been examined using the TG and the amine-modified tannin gel (ATG) prepared with ammonia treatment of the TG. The adsorption amounts of boron for the TG and the ATG were relatively small and almost constant below pH 7, whereas the boron adsorption amounts increased with increasing pH in the range of pH above 7. Considering that in aqueous solutions above pH 7, the mole fraction of boric acid decreases while that of tetrahydroxyborate ion increases with increasing pH, the boron adsorption onto both gels takes place probably through the chelate formation of tetrahydroxyborate ion with the hydroxy and the amino groups in the gels. Besides, the adsorbability of the ATG for boron was higher than that of the TG due to the stable coordination bond between boron and nitrogen of the amino group in the ATG. The adsorption kinetics were adequately described by the pseudo second order kinetic equation while the adsorption isotherms followed both the Langmuir and the Freundlich equations. The boron adsorbability of both the TG and the ATG at low boron concentration were comparable or fairly good compared with other adsorbents. PMID- 21645918 TI - Changes in D-dimer levels after cesarean section in women with singleton and twin pregnancies. AB - OBJECTIVE: To determine how the D-dimer level changes after cesarean section, a topic that has not been studied extensively. METHODS: The D-dimer level (MUg/mL) was measured in 792 and 119 women with singleton and twin pregnancies, respectively, on the day or one day before cesarean section and on postpartum days 1, 3, and 7. None of the women developed venous thromboembolism with clinical symptoms. The data for other two women who underwent a cesarean section and developed pulmonary thromboembolism on postpartum days 0 to 1 were also presented. RESULTS: The preoperative D-dimer level (median, MUg/mL) of 2.4 increased on postpartum day 1 to 6.0, then decreased to 2.8 on postpartum day 3, and again increased to 4.5 on postpartum day 7 in the singleton pregnancies. A similar pattern, but with slightly higher values, was seen in the twin pregnancies. The 95 percentile value of D-dimer for singleton and twin pregnancies was 6.9 and 10.5 on days -1/0, 19.7 and 25.7 on day 1, 9.7 and 13.5 on day 3, and 15.7 and 17.7 on day 7, respectively. The D-dimer level after pulmonary thromboembolism was greater than the 99 percentile value and the 98 percentile value in the two women, respectively. CONCLUSION: Our data regarding the D-dimer level may be helpful when considering the normal range of D-dimer for postpartum women with cesarean delivery. PMID- 21645919 TI - Tunable dual growth factor delivery from polyelectrolyte multilayer films. AB - A promising strategy to accelerate joint implant integration and reduce recovery time and failure rates is to deliver a combination of certain growth factors to the integration site. There is a need to control the quantity of growth factors delivered at different times during the healing process to maximize efficacy. Polyelectrolyte multilayer (PEM) films, built using the layer-by-layer (LbL) technique, are attractive for releasing controlled amounts of potent growth factors over a sustained period. Here, we present PEM films that sequester physiological amounts of osteogenic rhBMP-2 (recombinant human bone morphogenetic protein-2) and angiogenic rhVEGF165 (recombinant human vascular endothelial growth factor) in different ratios in a degradable [poly(beta-amino ester)/polyanion/growth factor/polyanion] LbL tetralayer repeat architecture where the biologic load scaled linearly with the number of tetralayers. No burst release of either growth factor was observed as the films degraded. The release of rhBMP-2 was sustained over a period of 2 weeks, while rhVEGF165 eluted from the film over the first 8 days. Both growth factors retained their efficacy, as quantified with relevant in vitro assays. rhBMP-2 initiated a dose dependent differentiation cascade in MC3T3-E1S4 pre-osteoblasts while rhVEGF165 upregulated HUVEC proliferation, and accelerated closure of a scratch in HUVEC cell cultures in a dose dependent manner. In vivo, the mineral density of ectopic bone formed de novo by rhBMP-2/rhVEGF165 PEM films was approximately 33% higher than when only rhBMP-2 was introduced, with a higher trabecular thickness, which would indicate a decrease in the risk of osteoporotic fracture. Bone formed throughout the scaffold when both growth factors were released, which suggests more complete remodeling due to an increased local vascular network. This study demonstrates a promising approach to delivering precise doses of multiple growth factors for a variety of implant applications where control over spatial and temporal release profile of the biologic is desired. PMID- 21645920 TI - Self-protecting core-shell magnetic nanoparticles for targeted, traceable, long half-life delivery of BCNU to gliomas. AB - The successful delivery of anti-cancer drugs relies on the simultaneous capability to actively target a specific location, a sufficient lifetime in the active form in the circulation, and traceability and quantification of drug distribution via in vivo medical imaging. Herein, a highly magnetic nanocarrier (HMNC) composed of an Fe(3)O(4) core and an aqueous-stable, self-doped poly[N-(1 one-butyric acid)]aniline (SPAnH) shell was chemically synthesized. This nanocarrier exhibited a high capacity for 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) drug loading. BCNU and o-(2-aminoethyl)polyethylene glycol (EPEG) were covalently immobilized on the surface of the HMNC to form a self-protecting magnetic nanomedicine (i.e., SPMNM) that could simultaneously provide low reticuloendothelial system uptake, high active-targeting, and in vivo magnetic resonance imaging (MRI) traceability. Meanwhile, the SPMNM was found to reduce the phagocytosis by macrophages and reduce the hydrolysis rate of BCNU. The high magnetization (approximately 1.2-fold higher than Resovist) of the HMNC allowed efficient magnetic targeting to the tumor. The synergetic drug delivery approach provided approximately a 3.4-fold improvement of the drug's half-life (from 18 h to 62 h) and significantly prolonged the median survival rate in animals that received a low dose of BCNU, compared with those that received a high dose of free BCNU (63 days for those that received 4.5 mg BCNU/kg carried by the nanocarrier versus 50 days for those that received 13.5 mg of free-BCNU). This improvement could enhance the potential of magnetic targeting therapy in clinical applications of cancer treatments. PMID- 21645921 TI - Involvement of osteopontin and its signaling molecule CD44 in clinicopathological features of adult T cell leukemia. AB - We previously reported that the osteopontin (OPN) gene as well as CD44 is trans activated by the Tax protein of HTLV-1, however the synthesis of both in adult T cell leukemia (ATL) has not been described yet. Here we showed the expression of these molecules in plasma and tissue of ATL. Significant differences were found among the normal and four subtypes of 27 ATL patients in plasma levels of OPN (p=3.6*10(-6)) and soluble CD44 (p<0.001) and they were significantly related to each other (p<0.002). Also they were significantly associated with the performance status, total number of involved lesions, and lactic dehydrogenase, and inversely with lymphocyte count (p<0.01). Immunohistochemical staining of lymph-nodes and skin from 7 ATL patients using anti-OPN and anti-CD44 antibodies demonstrated that both expressions were weak/moderate in ATL cells but moderate/strong in infiltrated macrophages in 6 patients. These results demonstrate that OPN and CD44 play important roles in tumor formations and their products in plasma could be markers of the severity in ATL. PMID- 21645923 TI - Being practical, being safe: doing evaluations in contested spaces. AB - This final article summarizes and synthesizes the full special edition. This volume questioned evaluation as philosophy, enterprise, and practice from the point of view of doing evaluation under conditions of social violence, disruption, and division. In this final article, we clarify the issues and problems which challenge the professional practice of evaluation and propose guiding questions for evaluators designing and doing evaluation in worlds like these. We attend to the consequences for evaluation and evaluator of choosing to inhabit these violent worlds as an evaluator. PMID- 21645922 TI - Factors associated with caregiver stability in permanent placements: a classification tree approach. AB - OBJECTIVE: Identify individual and environmental variables associated with caregiver stability and instability for children in diverse permanent placement types (i.e., reunification, adoption, and long-term foster care/guardianship with relatives or non-relatives), following 5 or more months in out-of-home care prior to age 4 due to substantiated maltreatment. METHODS: Participants were 285 children from the Southwestern site of Longitudinal Studies of Child Abuse and Neglect (LONGSCAN). Caregiver instability was defined as a change in primary caregiver between ages 6 and 8 years. Classification and regression tree (CART) analysis was used to identify the strongest predictors of instability from multiple variables assessed at age 6 with caregiver and child reports within the domains of neighborhood/community characteristics, caregiving environment, caregiver characteristics, and child characteristics. RESULTS: One out of 7, or 14% of the 285 children experienced caregiver instability in their permanent placement between ages 6 and 8. The strongest predictor of stability was whether the child had been placed in adoptive care. However, for children who were not adopted, a number of contextual factors (e.g., father involvement, expressiveness within the family) and child characteristics (e.g., intellectual functioning, externalizing problem behaviors) predicted stability and instability of permanent placements. CONCLUSIONS: Current findings suggest that a number of factors should be considered, in addition to placement type, if we are to understand what predicts caregiver stability and find stable permanent placements for children who have entered foster care. These factors include involvement of a father figure, family functioning, and child functioning. PRACTICE IMPLICATIONS: Adoption was supported as a desired permanent placement in terms of stability, but results suggest that other placement types can also lead to stability. In fact, with attention to providing biological parents, relative, and non-relative caregivers with support and resources (e.g., emotional, financial, and optimizing father involvement or providing a stable adult figure) the likelihood that a child will have a stable caregiver may be increased. PMID- 21645924 TI - Interleukin(IL)-4 promotion of CXCL-8 gene transcription is mediated by ERK1/2 pathway in human pulmonary artery endothelial cells. AB - Interleukin-4 is central to allergic pulmonary inflammatory responses, but its contribution to airway neutrophilia remains controversial. The endothelium plays a critical role in regulating leukocyte recruitment and migration during inflammation. However, its response to IL-4 is reported to either increase or decrease the production of neutrophil chemotactic factors. We hypothesized that these conflicting findings may be due to the tissue and the size of the vessels from which endothelial cells have been derived. The expression of CXCL-8 by human primary culture umbilical veins endothelial cells (HUVECs), human pulmonary artery endothelial cells (HPAECs), and human pulmonary microvascular endothelial cells (HPMECs) when stimulated with recombinant human IL-4 (rhIL-4) was studied. The chemoattractant property of the cells' supernatants for neutrophils was evaluated using Boyden chambers. The role of the nuclear factor-kappaB (NF kappaB), and mitogen-activated protein kinases (MAPK) in IL-4-induced HPAECs was studied using Western blotting and electrophoretic mobility shift assay (EMSA). We demonstrated that IL-4 increased the mRNA expression and the protein production of CXCL-8 in HPAECs, but not in HUVECs and HPMECs. The supernatants of HAPECs stimulated by IL-4 significantly promoted neutrophils migration in a dose dependent manner, and was significantly attenuated by an inhibitor of CXCL-8. We also found that extracellular-regulated protein kinase1/2 (ERK1/2) is activated by IL-4 in HPAECs, but not JUN-N-terminal protein kinase (JNK) or p38 MAPK pathway. Furthermore, NF-kappaB-DNA binding activity, phosphorylation of IkappaBalpha and p65 levels were not affected by rhIL-4 in HAPECs. These findings indicate marked functional differences in the response of micro and macro-ECs to IL-4. ERK1/2, rather than NF-kappaB, JNK and p38 MAPK signaling, plays a role in IL-4 induced chemokine activation. Our results suggest that inhibition of ERK1/2 may be a possible target for airway neutrophilia in allergic lung diseases. PMID- 21645925 TI - Plasticity of human CD8alphaalpha binding to peptide-HLA-A*2402. AB - The human CD8 functions as a co-receptor for specific T cell recognition, and only one complex structure of human CD8alphaalpha binding to HLA-A*0201 has been solved, revealing the molecular basis of CD8 interacting with its ligand pHLA. Here, we present the complex structures of human CD8alphaalpha bound to HLA A*2402, which demonstrate two opposite alpha3 domain CD loop shifts (either pull or push) in the HLA heavy chain upon CD8 engagement. Taking the previously reported mouse CD8-pMHC complex structures into account, from the structural view, all of the data indicate the plasticity of CD8 binding to pMHC/HLA, which facilitates its co-receptor function for T cells. The plasticity of CD8 binding appears not to affect the specificity of TCR recognition, as no peptide conformation change extends to the pMHC interface for TCR contacting. PMID- 21645926 TI - Novel method for differentiation between Trastuzumab and host adaptive response. AB - Humoral immune response to human epidermal growth factor receptor 2 (HER-2/neu or ErbB-2) has been detected in sera of breast cancer patients and shown to be an appropriate prognostic marker (Taylor et al., 2007). However, since Trastuzumab (Herceptin) is a widely used monoclonal antibody as cancer therapy agent for tumors over-expressing HER-2, there is a need for an efficient way to detect host generated antibodies against HER-2 without the confounding effect of Herceptin. Here we describe a screening method developed to decipher between host antibodies against HER-2 and that of Herceptin. By producing a series of truncation mutants within the epitope of Herceptin, we were able to inhibit this binding. We demonstrated also that by a three amino acid substitution (PPF->SSS) we were able to abrogate Herceptin binding while generating a highly conserved HER-2 extracellular domain (ECD). By producing a stable cell line that expresses this mutated form of the human HER-2 ECD, we have a source of this protein to probe patient sera. Our method represents a proof of principle that mutated HER-2 which we constructed could be used to distinguish between a host response against HER-2 and the monoclonal antibody Herceptin targeting the same protein. PMID- 21645927 TI - Blastocyst-mediated induction of endometrial connexins: an inflammatory response? AB - As a prerequisite for successful embryo implantation in mammals, before implantation ovarian hormones regulate the transformation of the endometrium into the receptive phase. During the implantation process, gene expression in the receptive endometrium is additionally modulated by the presence of a blastocyst. During this complex differentiation process, in humans as well as in rodents, gap junction connexin 26 (Cx26) is suppressed in the uterine epithelium and Cx43 is suppressed in the endometrial stromal cells during the receptive phase. In rodents, a blastocyst-mediated induction of Cx26 takes place locally in the uterine epithelial cells of the implantation chamber surrounding the blastocyst, followed by an increase in Cx43 in the cells of the developing decidua. The Cx26 induction is dependent on the presence of a blastocyst and occurs even before adhesion and invasion of the trophoblast takes place. The signal cascades involved in this blastocyst-mediated connexin induction are still elusive. The process of implantation is considered as a proinflammatory response, and inflammatory factors have been shown to be involved in the implantation process. In fact, Cx26 expression can be induced in the receptive rat endometrium by mediators of the inflammatory cascade including prostaglandin-F2alpha and IL1beta by an ER-independent pathway similar to the blastocyst-mediated connexin induction at the time of implantation. Thus, in the receptive endometrium induction of connexin expression may also be induced by mediators of the inflammatory signaling cascade, and the implantation-related induction of intercellular communication may in part be due to an inflammatory response. PMID- 21645928 TI - Future directions of studies for recurrent miscarriage associated with immune etiologies. AB - A significant proportion of recurrent pregnancy loss (RPL) is associated with immune etiologies. The immunological environment is different between decidua basalis and decidua parietalis, and also different between RPL cases with normal fetal chromosomes and those with abnormal fetal chromosomes. Recent data show that the immune system in a late-stage abortion is completely different from that in an early-stage abortion. If immunocompetent cells can cause RPL, the immunological environment may be a causative factor, especially in an early-stage abortion, and/or at decidua basalis and/or in the cases of RPL with a normal embryo. Careful examination of the immune system at the decidua basalis in an early-stage abortion in RPL cases with normal fetal chromosome may reveal useful information. PMID- 21645929 TI - Approach and avoidance motivation in eating disorders. AB - It has been proposed that approach and avoidance processes may be critically involved in the development and maintenance of eating disorders (EDs), including anorexia nervosa (AN) and bulimia nervosa (BN). The Behavioural Inhibition System and Behavioural Activation System Scales (BIS/BAS) and Appetitive Motivation Scale (AMS) questionnaires were administered to 286 participants: 91 healthy controls (HCs), 121 participants with a current ED, either AN (restrictive and binge purge subtypes), or BN and 74 participants recovered from an ED. Individuals with EDs had higher levels of sensitivity to punishment and lower levels of reward reactivity than controls. Individuals in recovery from an ED scored the same as those in the acute group, with the exception of BAS fun seeking, for which they scored significantly higher than those with restricting AN. Discriminant analysis revealed that HCs were maximally separated from those in the acute and recovered ED groups along a dimension reflecting high punishment sensitivity and low reward sensitivity. Classification analysis demonstrated that ED and HC group membership was predicted from reward and punishment sensitivity measures; however recovered participants tended to be misclassified as ED. This study suggests high punishment sensitivity and low reward reactivity/sensitivity might form a personality cluster associated with the risk of developing an ED. PMID- 21645930 TI - Recombinant canine granulocyte colony-stimulating factor accelerates recovery from cyclophosphamide-induced neutropenia in dogs. AB - In dogs injected intravenously with 400mg/m(2) cyclophosphamide (CPA), the peripheral neutrophil count decreased to less than 1000 cells/MUL in 5-9 days. Treatment with purified recombinant canine granulocyte colony-stimulating factor (rcG-CSF), produced by brevibacillus expression system, at the nadir of the granulocyte count accelerated recovery from the CPA-induced neutropenia by 1-3 days. Therapeutic administration of rcG-CSF at doses of 2.5-10 MUg/kg did not show any significant difference on the severity of neutropenia (the period that granulocyte counts were less than 2000 cells/MUL). Administration of 2.5 MUg/kg rcG-CSF 3 times per day 2-4 days or 3-5 days after CPA treatment not only accelerated recovery but also decreased the severity of neutropenia. No clinical signs of the rcG-CSF were observed. These results showed that the rcG-CSF is effective for treatment of neutropenia in dogs. PMID- 21645931 TI - Heterophil functional responses to dietary immunomodulators vary in genetically distinct chicken lines. AB - The effect of dietary supplementation of immunomodulators on in vitro chicken heterophil function was investigated using three diverse genetic lines of chickens (broiler, Fayoumi, and Leghorn). Dietary supplementation with beta glucan, ascorbic acid, and corticosterone was fed from 8 to 11 weeks of age. Heterophil function was evaluated weekly during supplementation using phagocytosis, bacterial killing, and heterophil extracellular traps (HETs)-DNA release. Fayoumis fed the basal diet had significantly higher HETs-DNA release (P=0.002) than Leghorns and broilers. Both genetic line and immunomodulator diet supplementation had significant effects on bacterial killing (line and diet effect: P<0.001) and HETs-DNA release (line: P<0.001; diet: P=0.043). Dietary supplementation with immunomodulators, therefore, shows potential to affect and augment heterophil function in chickens. The current results also suggest the important role of genetics in innate immune responses. PMID- 21645932 TI - Seizure-induced Takotsubo syndrome is more frequent than reported. PMID- 21645933 TI - Blood pressure-dependent and independent effects of amlodipine and angiotensin receptor blockers on the risk of stroke. PMID- 21645934 TI - Post-mortem examination and laboratory-based analysis for the diagnosis of bovine tuberculosis among dairy cattle in Ecuador. AB - Veterinary inspection in slaughterhouses allows for the detection of macroscopic lesions reminiscent of bovine tuberculosis, but the presence of Mycobacterium bovis must be confirmed by laboratory methods. This study aimed at comparing the performances of the standard diagnostic tools used to identify M. bovis in tissue specimens sampled from suspicious animals. During a two years period, 1390 cattle were inspected at the Machachi abattoir in the Mejia canton - Ecuador. A total of 33 animals with granulomatous lesions were detected, representing 2.33% (16/687) and 2.42% (17/703) animals examined in 2007 and 2008, respectively. Ninety-four tissue specimens were sampled and screened for the presence of mycobacteria. Acid fast bacilli were identified in one third of the suspicious cattle (11/33) and suggestive microscopic lesions in 27.3% (9/33) of the samples examined by direct microscopy and histopathology, respectively. Culturing on Stonebrink medium and 16S-rRNA-based polymerase chain reaction (PCR) yielded 36.4% (12/33) and 27.3% (9/33) of positives, respectively. Compared to culture, other diagnostic procedures displayed a lower sensitivity, with 56.5% for PCR, and 43.5% for direct microscopy and histopathology; however, the specificity was higher (94.4% for PCR and microscopy, and 97.2% for histopathology). We conclude that reliable post-mortem laboratory testing either requires the combination of a set of available diagnostic tools or necessitates the development of improved new generation tools with better sensitivity and specificity characteristics. PMID- 21645935 TI - Effect of breeding strategy and feeding system on the within-herd variation of lean meat percents in Danish slaughter pigs. AB - In several countries slaughter pigs are paid for individually, according to slaughter weight and lean meat percent (LMP). Production of uniform batches of pigs within the optimal weight and LMP limits will obtain the best price. Therefore, all pigs should have a similar growth rate (average daily gain, ADG) and reach an appropriate slaughter weight within the same time period. LMP may serve as a proxy for ADG since pigs with low LMP have significantly higher ADG than pigs with high LMP and vice versa. Both breeding strategy and feeding system may influence the range of variation among pigs. The aim of this study was to test the two following hypotheses: (1) Herds purchasing breeding gilts have a higher mean value and a lower variation (standard deviation) in LMP than herds producing their own breeding gilts and (2) Herds using restricted feeding of finishers have a higher mean value and a lower variation (standard deviation) in LMP than herds with ad libitum feeding of finishers. The study included 72 herds and a total of 345,132 pigs slaughtered during one year. Among the 72 herds, 40 were home-breeders and 32 purchased breeding gilts from a breeding company. Nineteen herds used restricted feeding, of which 8 (42%) were home-breeders. Fifty-three herds used ad libitum feeding, of which 32 (60%) were home-breeders. Breeding strategy had a significant effect on SDLMP (p=0.003), where purchase of breeding gilts resulted in a significantly lower standard deviation of the monthly LMP compared to home-bred gilts (a difference in median SDLMP of 0.2 percentage points or 8% difference between groups). Feeding system had a significant effect on the meanLMP (p<0.001), with a significantly higher meanLMP in herds using restrictive feeding compared to ad libitum feeding (60.7% versus 60.0%). Restrictive feeding also resulted in a significantly lower SDLMP (p<0.001) compared to ad libitum feeding (2.2% versus 2.5% or a 12% difference between groups). PMID- 21645936 TI - Seroprevalence of Q fever in naturally infected dairy cattle herds. AB - Coxiella burnetii is the causal agent of Q fever, a worldwide spread zoonosis. Prevention of C. burnetii shedding in cattle is critical to control the spread of the pathogen between animals, and from animals to humans. Vaccination with a phase 1 vaccine has been shown to be effective in preventing shedding when implemented in still susceptible animals, even in infected cattle herds. The identification of these animals (dairy cows and nulliparous females) as targets for vaccination consequently is crucial. Hygiene measures conventionally also are implemented, but their relative impact on C. burnetii diffusion remains unknown. The objectives of this study therefore were to (i) describe the distribution of the within-herd apparent seroprevalence among cows and nulliparous females and (ii) to explore the association between management practices and herd characteristics on the one hand, and these seroprevalences on the other. In a sample of 100 naturally and clinically infected dairy herds, blood samples were taken systematically from all nulliparous females (older than 12 months) and cows, and serologically tested. Information on herd characteristics and management practices were collected through a questionnaire filled in by each farmer. The variation in within-herd seroprevalence among cows and the risk for a herd of having at least one seropositive nulliparous female were investigated using multivariate (linear and logistic respectively) regression models. Median within-herd seroprevalence was 0.32 (Q1=0.22; Q3=0.43). We observed a low to null (median=0.01; Q1=0; Q3=0.10) within-herd seroprevalence in nulliparous females contrary to a high value (median=0.42) and variability (Q1=0.28; Q3=0.56) in cows. Only a few herd characteristics and management practices were found to be related to seroprevalence. Within-herd seroprevalence in cows was found to be significantly (P<0.10) higher in herds (i) with a number of cows<46, (ii) with seasonal calving, and (iii) with grazing or contact through the fence with other ruminant herds. The risk of having at least one seropositive nulliparous female was increased in herds (i) with seasonal calving and (ii) where the foetus and/or the placenta of aborted cows were not systematically removed. Our findings support, in addition to the implementation of high level of hygiene measures, the relevance of vaccination (at least in nulliparous females) as a method to control the spread of C. burnetii within an infected herd, as vaccination is effective in susceptible animals and given that nulliparous females are mostly not infected even in infected herds. PMID- 21645937 TI - Anti-Neospora caninum antibodies in milk in relation to production losses in dairy cattle. AB - A comprehensive field study was carried out with the following objectives: (a) to assess the usefulness of individual and bulk tank milk analysis for determining Neospora caninum serostatus in individual cows and herds, and (b) to study the associations between N. caninum infection status (based on milk testing), and several productive and reproductive parameters in the animals. Antibodies were detected with a commercially available ELISA test (Bio K 192/5). Analysis of paired serum and milk samples from 1134 lactating cows on 38 farms revealed that 97.6% of the ELISA results were coincident, irrespective of whether serum or milk samples were used. Moreover, multiple linear regression analysis revealed that 86.0% of the variations in ELISA values in milk were due to variations in the serum. The measurement of antibodies in bulk tank milk was a good estimator of the herd level status of N. caninum infection, and enabled detection of infection in 94.7% herds with >=10.0% seropositive cows and/or in all herds with >4% highly seropositive cows. The odds ratio for abortion in seropositive animals was 9.1 times higher than in seronegative animals. The infection serostatus was also a significant risk factor, as the odds ratio for abortion was even higher (12.0 times) in cows categorized as highly seropositive. ELISA values for the bulk milk from 387 randomly selected herds were negatively associated with average milk production. Moreover, milk production losses mainly occurred on farms categorized as highly positive (i.e. herds with >=20.0% seropositive cows). PMID- 21645938 TI - Mimicking natural bio-mineralization processes: a new tool for osteochondral scaffold development. AB - In recent years, the concept of regenerative medicine has gained great importance, particularly in the field of orthopaedics, in which current solutions are based mainly on the replacement of damaged tissues with devices that function only as structural replacements with limited regenerative capacity. New regenerative solutions can be obtained by taking inspiration from nature, which surrounds us with a multitude of organisms endowed with extraordinary performance. In particular, bio-mineralization, which is the basis of the formation of load-bearing structures in vertebrate and invertebrate organisms, can be exploited to achieve innovative devices for the repair and reconstruction of bone and osteo-cartilaginous tissues. PMID- 21645939 TI - Metabolomes of the psychrotolerant bacterium Listeria monocytogenes 10403S grown at 37 degrees C and 8 degrees C. AB - Listeria monocytogenes is a food-borne pathogen with the ability to grow at refrigeration temperatures. Knowledge of the mechanisms involved in low temperature growth is incomplete and here we report the results of a metabolomics investigation of this. The small molecule contents of L. monocytogenes 10403S grown at 37 degrees C and 8 degrees C were compared by gas chromatography/mass spectrometry (GC/MS). Over 500 peaks were detected in both 37 degrees C and 8 degrees C-grown cells, and 103 were identified. Of the identified metabolites, the concentrations of 56 metabolites were increased (P<0.05), while the concentrations of 8 metabolites were decreased at low temperature. Metabolites increasing in concentration at 8 degrees C included amino acids, sugars, organic acids, urea cycle intermediates, polyamines, and different compatible solutes. A principal component analysis (PCA) was used to visualize and compare the matrix containing the data in 6 samples, and this clearly identified the 37 degrees C and 8 degrees C metabolomes as different. The results indicated that an increase in solute concentrations in the cytoplasm was associated with low temperature adaptation, which may be a response to chill stress with the effect of lowering the freezing point of intracellular water and decreasing ice crystal formation. PMID- 21645940 TI - The rs1801278 G>A polymorphism of IRS-1 is associated with metabolic syndrome in healthy nondiabetic men. Modulation by cigarette smoking status. AB - AIMS: To explore associations between IRS-1 rs1801278 G>A polymorphism and metabolic syndrome (MS). METHODS: rs1801278 G>A was genotyped in 610 healthy Argentinian men. RESULTS: GA carriers had lower risk of MS (OR=0.52, P=0.045), particularly among smokers (OR=0.10, P=0.006). CONCLUSIONS: rs1801278 GA carriers had lower risk of MS, especially among smokers. PMID- 21645941 TI - Averaging in vitro cardiac field potential recordings obtained with microelectrode arrays. AB - Extracellular field potential (FP) recordings with microelectrode arrays (MEAs) from cardiomyocyte cultures offer a non-invasive way of studying the electrophysiological properties of these cells at the population level. Several studies have examined the FP properties of cardiomyocytes of various origins, including stem cell-derived cardiomyocytes. This focus reflects growing importance and interest in the field of MEA. High-quality cardiac FP signals are often difficult to obtain, especially from stem cell-derived cardiomyocyte cultures, which represent an important new field in cardiac electrophysiology. One way to improve the quality of these recordings is to average the cardiac FP signals. To date, however, no studies have examined the effect of averaging on cardiac FP signals. We report here that cardiac FP averaging can yield higher quality signals than original individual FPs, and therefore promise more accurate detection of different phases and analysis of the cardiac FP signal. Averaged signals improved the signal-to-noise ratio (SNR), and obtaining reliable averages required approximately 50 cardiac cycles. We therefore propose that routine cardiac FP averaging can serve as a tool to compare the effects of different experimental conditions or stimuli on the properties of cardiac FPs. PMID- 21645942 TI - Association of smoking with tumor size at diagnosis in non-small cell lung cancer. AB - Tumor size at diagnosis (TSD) indirectly reflects tumor growth rate. The relationship between TSD and smoking is poorly understood. The aim of the study was to determine the relationship between smoking and TSD. We reviewed 1712 newly diagnosed and previously untreated non-small cell lung cancer (NSCLC) patients' electronic medical records and collected tumor characteristics. Demographic and epidemiologic characteristics were derived from questionnaires administered during personal interviews. Univariate and multivariate linear regression models were used to evaluate the relationship between TSD and smoking controlling for demographic and clinical factors. We also investigated the relationship between the rs1051730 SNP in an intron of the CHRNA3 gene (the polymorphism most significantly associated with lung cancer risk and smoking behavior) and TSD. We found a strong dose dependent relationship between TSD and smoking. Current smokers had largest and never smokers smallest TSD with former smokers having intermediate TSD. In the multivariate linear regression model, smoking status (never, former, and current), histological type (adenocarcinoma versus SqCC), and gender were significant predictors of TSD. Smoking duration and intensity may explain the gender effect in predicting TSD. We found that the variant allele of rs1051730 in CHRNA3 gene was associated with larger TSD of squamous cell carcinoma. In the multivariate linear regression model, both rs1051730 and smoking were significant predictors for the size of squamous carcinomas. We conclude that smoking is positively associated with lung tumor size at the moment of diagnosis. PMID- 21645943 TI - A systematic review of morphea treatments and therapeutic algorithm. AB - BACKGROUND: Morphea (localized scleroderma) is a skin disorder with significant morbidity. No consistent recommendations exist for therapy, impeding patient care. OBJECTIVE: We sought to create an evidence-based therapeutic algorithm. METHODS: We reviewed English-language literature using search engines and hand searches for therapeutic interventions in morphea. Results were summarized. RESULTS: Narrowband ultraviolet B is appropriate for progressive or widespread superficial dermal lesions; broadband ultraviolet A/ultraviolet A-1 is appropriate for widespread or progressive deeper dermal lesions. Systemic treatment with methotrexate, corticosteroids, or both is indicated for deep or function-impairing lesions and rapidly progressive or widespread (severe) disease. Topical treatment with calcipotriene or tacrolimus is supported for limited, superficial, inflammatory lesions. Use of oral calcipotriol, D penicillamine, interferon gamma, and antimalarials is not supported. LIMITATIONS: Limitations are publication bias; lack of adequately powered, controlled trials; and no validated outcome measures. CONCLUSION: Phototherapy, methotrexate/systemic corticosteroids, calcipotriene, and topical tacrolimus have the most evidence for efficacy in morphea. Treatment works best in inflammatory disease. Disease activity, severity, progression, and depth should play a role in therapeutic decision making. PMID- 21645944 TI - [Spontaneous bacterial empyema: a note on terminology]. PMID- 21645945 TI - Can an intervention on clinical inertia have an impact on the perception of pain, functionality and quality of life in patients with hip and/or knee osteoarthritis? Results from a cluster randomised trial. AB - OBJECTIVES: Evaluate whether an intervention applied to general practitioners to prevent clinical inertia had an impact on pain, functionality, and health-related quality of life (HRQoL) of patients with hip and/or knee osteoarthritis. DESIGN: This was a cluster-based, multicentre, prospective, randomized, parallel-group study. Clusters of physicians working were assigned to one of two study groups. Physicians in Group 1 received a training session while those in Group 2 did not. SETTING: Primary Care Health centers representative of the entire Spanish territory. PARTICIPANTS: 329 general practitioners of primary healthcare centre. INTERVENTIONS: The intervention consists of a motivational session to propose a proactive care, based on current recommendations. MEASUREMENTS: Visual analogue scale (VAS); functionality (WOMAC scale) and global perception of health by SF 12. Effects were measured in two visits six months apart. RESULTS: A total of 1361 physicians, and 4076 patients participated in the study. No significant differences were observed in the clinical benefit obtained between patients assigned to Group 1 and Group 2. Nevertheless, a significant improvement was observed in the combined population (Groups 1 + 2) in the VAS (p<0.001), WOMAC (p<0.0001) and SF-12v2 (p<0.001) questionnaires in Visit 2 compared to Visit 1. CONCLUSIONS: The results indicate that, although this specific intervention carried out on physicians did not provide an additional clinical benefit to patients with knee and/or hip osteoarthritis, an increased awareness of the patient's disease through the use of functionality indexes, as well as the mere fact of being observed, seem to improve patient-reported pain, functionality and HRQoL. PMID- 21645946 TI - Synthesis and anticancer activity of (RS)-9-(2,3-dihydro-1,4-benzoxaheteroin-2 ylmethyl)-9H-purines. AB - Herein are reported the synthesis and anticancer activity against the human breast cancer cell line MCF-7 of a series of substituted (RS)-9-(2,3-dihydro-1,4 benzoxathiin-2-ylmethyl)-9H-purine derivatives and (RS)-9-(2,3-dihydro-1,4 benzodioxin-2-ylmethyl)-9H-purine derivatives. When the Mitsunobu reaction was carried out between (RS)-2,3-dihydro-1,4-benzoxathiin-3-methanol and the heterocyclic bases 6-chloro-, 2,6-dichloro, and 6-bromo-purines under microwave assisted conditions, a formal 1,4-sulfur migration takes place through two consecutive oxyranium and episulfonium rings, giving rise to the corresponding (RS)-9-(2,3-dihydro-1,4-benzodioxin-3-ylmethyl)-9H-purine derivatives, previously reported by us. The most active compound (RS)-2,6-dichloro-9-(2,3-dihydro-1,4 benzoxathiin-2-ylmethyl)-9H-purine shows an IC(50) = 2.75 +/- 0.02 MUM. When the cancerous cells were treated with this compound, a significant increase of apoptotic cells (70.08 +/- 0.33%) was obtained in relation to the control ones. The induction of the G(2)/M cell cycle arrest and apoptosis by the three most active compounds is associated with increased phosphorylation of eIF2alpha in human breast cancer cells. PMID- 21645947 TI - Students' experiences of blended learning across a range of postgraduate programmes. AB - The article describes the students' experiences of taking a blended learning postgraduate programme in a school of nursing and midwifery. The indications to date are that blended learning as a pedagogical tool has the potential to contribute and improve nursing and midwifery practice and enhance student learning. Little is reported about the students' experiences to date. Focus groups were conducted with students in the first year of introducing blended learning. The two main themes that were identified from the data were (1) the benefits of blended learning and (2) the challenges to blended learning. The blended learning experience was received positively by the students. A significant finding that was not reported in previous research was that the online component meant little time away from study for the students suggesting that it was more invasive on their everyday life. It is envisaged that the outcomes of the study will assist educators who are considering delivering programmes through blended learning. It should provide guidance for further developments and improvements in using Virtual Learning Environment (VLE) and blended learning in nurse education. PMID- 21645948 TI - Temporal genetic variability and host sources of Escherichia coli associated with fecal pollution from domesticated animals in the shellfish culture environment of Xiangshan Bay, East China Sea. AB - This study was conducted to analyze the genetic variability of Escherichia coli from domesticated animal wastes for microbial source tracking (MST) application in fecal contaminated shellfish growing waters of Xiangshan Bay, East China Sea. (GTG)(5) primer was used to generate 1363 fingerprints from E. coli isolated from feces of known 9 domesticated animal sources around this shellfish culture area. Jackknife analysis of the complete (GTG)(5)-PCR DNA fingerprint library indicated that isolates were assigned to the correct source groups with an 84.28% average rate of correct classification. Based on one-year source tracking data, the dominant sources of E. coli were swine, chickens, ducks and cows in this water area. Moreover, annual and spatial changes of E. coli concentrations and host sources may affect the level and distribution of zoonotic pathogen species in waters. Our findings will further contribute to preventing fecal pollution in aquatic environments and quality control of shellfish. PMID- 21645949 TI - Atmospheric phenanthrene pollution modulates carbon allocation in red clover (Trifolium pratense L.). AB - The influence of atmospheric phenanthrene (PHE) exposure (160 MUg m(-3)) during one month on carbon allocation in clover was investigated by integrative (plant growth analysis) and instantaneous (13)CO(2) pulse-labelling approaches. PHE exposure diminished plant growth parameters (relative growth rate and net assimilation rate) and disturbed photosynthesis (carbon assimilation rate and chlorophyll content), leading to a 25% decrease in clover biomass. The root-shoot ratio was significantly enhanced (from 0.32 to 0.44). Photosynthates were identically allocated to leaves while less allocated to stems and roots. PHE exposure had a significant overall effect on the (13)C partitioning among clover organs as more carbon was retained in leaves at the expense of roots and stems. The findings indicate that PHE decreases root exudation or transfer to symbionts and in leaves, retains carbon in a non-structural form diverting photosynthates away from growth and respiration (emergence of an additional C loss process). PMID- 21645950 TI - Distribution and bioaccumulation of steroidal and phenolic endocrine disrupting chemicals in wild fish species from Dianchi Lake, China. AB - The distribution and bioaccumulation of steroidal and phenolic endocrine disrupting chemicals (EDCs) were studied in various tissues of wild fish species from Dianchi Lake, China. In muscle tissue, 4-tert-octylphenol, 4-cumylphenol, 4 nonlyphenol and bisphenol A were detected in fish from each sampling site, with maximal concentrations of 4.6, 4.4, 18.9 and 83.5 ng/g dry weight (dw), respectively. Steroids (estrone, 17beta-estradiol 17alpha-ethynylestradiol and estriol) were found at lower levels (<11.3 ng/g dw) and less frequently in muscle samples. The highest concentrations of steroids and phenols were found in liver, followed by those in gill and the lowest concentration was found in muscle. The field bioconcentration factors (BCFs) of phenols were calculated in fish species ranged from 18 to 97. Moreover, the measured tissue concentrations were utilized in order to estimate water concentration of steroids (4.4-18.0 ng/L). These results showed that steroidal and phenolic EDCs were likely ubiquitous contaminants in wild fish. PMID- 21645951 TI - Inherent mineralization of 2,6-dichlorobenzamide (BAM) in unsaturated zone and aquifers--effect of initial concentrations and adaptation. AB - The dichlobenil metabolite BAM (2,6-dichlorobenzamide) is frequently detected in aquifers e.g. in Denmark despite the mother compound dichlobenil was banned here since 1997. BAM mineralization was investigated at environmentally relevant concentrations in sediment samples. Undisturbed sediment cores with known dichlobenil application were collected from topsoil to 8.5 m below surface resulting in 57 samples hereof 4 aquifer samples. Mineralization was only substantial (>10%) in the uppermost meter of the unsaturated zone. Microbial adaptation, observed as faster mineralization in pre-exposed than in pristine sediments from the same location, was only evident in sandy sediment where dichlobenil was still present, but not in clayey sediments. Higher initial concentrations (1-5000 MUg/kg) did not stimulate mineralization in pristine clayey or sandy sediments, or in pre-exposed sand. However, in pre-exposed clay mineralization was stimulated at high concentrations. Furthermore BAM was for the first time mineralized in aerobic aquifer sediments from different BAM contaminated groundwater locations. PMID- 21645952 TI - Substituting missing data in compositional analysis. AB - Multivariate analysis of environmental data sets requires the absence of missing values or their substitution by small values. However, if the data is transformed logarithmically prior to the analysis, this solution cannot be applied because the logarithm of a small value might become an outlier. Several methods for substituting the missing values can be found in the literature although none of them guarantees that no distortion of the structure of the data set is produced. We propose a method for the assessment of these distortions which can be used for deciding whether to retain or not the samples or variables containing missing values and for the investigation of the performance of different substitution techniques. The method analyzes the structure of the distances among samples using Mantel tests. We present an application of the method to PCDD/F data measured in samples of terrestrial moss as part of a biomonitoring study. PMID- 21645953 TI - Dispersal networks for enhancing bacterial degradation in heterogeneous environments. AB - Successful biodegradation of organic soil pollutants depends on their bioavailability to catabolically active microorganisms. In particular, environmental heterogeneities often limit bacterial access to pollutants. Experimental and modelling studies revealed that fungal networks can facilitate bacterial dispersal and may thereby improve pollutant bioavailability. Here, we investigate the influence of such bacterial dispersal networks on biodegradation performance under spatially heterogeneous abiotic conditions using a process based simulation model. To match typical situations in polluted soils, two types of abiotic conditions are studied: heterogeneous bacterial dispersal conditions and heterogeneous initial resource distributions. The model predicts that networks facilitating bacterial dispersal can enhance biodegradation performance for a wide range of these conditions. Additionally, the time horizon over which this performance is assessed and the network's spatial configuration are key factors determining the degree of biodegradation improvement. Our results support the idea of stimulating the establishment of fungal mycelia for enhanced bioremediation of polluted soils. PMID- 21645954 TI - Infertility treatment and fertility-specific distress: A longitudinal analysis of a population-based sample of U.S. women. AB - Because research on infertile women usually uses clinic-based samples of treatment seekers, it is difficult to sort out to what extent distress is the result of the condition of infertility itself and to what extent it is a consequence of the experience of infertility treatment. We use the National Survey of Fertility Barriers, a two-wave national probability sample of U.S. women, to disentangle the effects of infertility and infertility treatment on fertility-specific distress. Using a series of ANOVAs, we examine 266 infertile women who experienced infertility both at Wave 1 and at Wave 2, three years later. We compare eight groups of infertile women based on whether or not they have received treatment and on whether or not they have had a live birth. At Wave 1, infertile women who did not receive treatment and who had no live birth reported lower distress levels than women who received treatment at Wave 1 only, regardless of whether their infertility episode was followed by a live birth. At Wave 2, women who received no treatment have significantly lower fertility specific distress than women who were treated at Wave 1 or at Waves 1 and 2, regardless of whether there was a subsequent live birth. Furthermore, fertility specific distress did not increase over time among infertile women who did not receive treatment. The increase infertility-specific distress was significantly higher for women who received treatment at Wave 2 that was not followed by a live birth than for women who received no treatment or for women who received treatment at Wave 1 only. These patterns suggest that infertility treatment is associated with levels of distress over and above those associated with the state of being infertile in and of itself. PMID- 21645955 TI - Full neurological recovery following cardiac arrest during percutaneous coronary intervention due to accidentally intracoronary administration of ajmaline. PMID- 21645957 TI - Severe dermatitis caused by diltiazem. PMID- 21645958 TI - Neutrophil elastase and interleukin-6 in amniotic fluid as indicators of chorioamnionitis and funisitis. AB - OBJECTIVE: We wished to assess the diagnostic value of amniotic fluid concentrations of neutrophil elastase and interleukin-6 concentrations for the rapid detection of chorioamnionitis and funisitis. STUDY DESIGN: A retrospective study of 56 women who underwent transabdominal amniocentesis within 48 h before preterm delivery or spontaneous abortion. Statistical analyses included Student's t-test and Tukey-Kamer's HSD test. Receiver operating characteristics (ROC) curves were drawn to assess the predictive performance of the two markers. RESULTS: Neutrophil elastase concentrations differed significantly between patients with chorioamnionitis and other stages of chorioamnionitis, while interleukin-6 concentrations did not. To predict chorioamnionitis, concentrations of 3563 ng/ml for neutrophil elastase and 11,279 pg/ml for interleukin-6 were optimal. Comparison of ROC curves showed that neutrophil elastase was a significantly more sensitive predictor of funisitis than interleukin-6. CONCLUSION: Amniotic fluid concentrations of neutrophil elastase are more sensitive than interleukin-6 for the rapid detection of chorioamnionitis and funisitis. PMID- 21645959 TI - A bifunctional allosteric site in the dimer interface of procaspase-3. AB - The dimer interface of caspase-3 contains a bifunctional allosteric site in which the enzyme can be activated or inactivated, depending on the context of the protein. In the mature caspase-3, the binding of allosteric inhibitors to the interface results in an order-to-disorder transition in the active site loops. In procaspase-3, by contrast, the binding of allosteric activators to the interface results in a disorder-to-order transition in the active site. We have utilized the allosteric site to identify a small molecule activator of procaspase and to characterize its binding to the protease. The data suggest that an efficient activator must stabilize the active conformer of the zymogen by expelling the intersubunit linker from the interface, and it must interact with active site residues found in the allosteric site. Small molecule activators that fulfill the two requirements should provide scaffolds for drug candidates as a therapeutic strategy for directly promoting procaspase-3 activation in cancer cells. PMID- 21645961 TI - Differential diagnosis between pre- and postganglionic adult traumatic brachial plexus lesions by ultrasonography. AB - The goal of this study was to prospectively investigate the feasibility of preoperative ultrasonography evaluation in the differentiation between pre- and postganglionic traumatic brachial plexus lesions. Two expert radiologists employed ultrasonography to observe the morphology of the brachial plexus in 23 patients with suspected traumatic brachial plexus lesions and 40 healthy volunteers. The detection rate was 100% (126/126) for the C5 through C7 nerve roots and upper and middle trunks and three fascicles, 84% (106/126) for the C8 roots and the lower trunks and 64% (81/126) for T1 roots in all subjects. Surgical inspections found 58 lesions in 23 patients (40 preganglionic lesions and 18 postganglionic lesions). Ultrasonography detected most of the brachial plexus lesions (56/58) but misjudged two preganglionic and two postganglionic lesions. The rate of differentiation was 93% (52/56). This study demonstrated that ultrasonography is a useful but experience-dependent supplemental imaging technique for preoperative diagnosis of brachial plexus lesions and differentiation between pre- and postganglionic brachial plexus lesions. PMID- 21645962 TI - Emerging role of ultrasonography in rheumatoid arthritis: optimizing diagnosis, measuring disease activity and identifying prognostic factors. AB - Ultrasonography is a sensitive imaging modality that provides valuable information regarding early inflammatory changes that are not detected by clinical examination or X-rays, such as subclinical synovitis and erosions. This information may improve the management of rheumatoid arthritis by providing a more timely and accurate diagnosis, identifying poor prognostic factors, more accurately monitoring response to therapeutic intervention, improving treatment decisions and more accurately assessing remission. Ultrasonography could play a critical role in minimizing disease activity through strict monitoring and aggressive therapeutic adjustment, which has emerged as an approach to improve long-term outcomes for patients with rheumatoid arthritis. PMID- 21645960 TI - PPARgamma: a molecular link between systemic metabolic disease and benign prostate hyperplasia. AB - The emergent epidemic of metabolic syndrome and its complex list of sequelae mandate a more thorough understanding of benign prostatic hyperplasia and lower urinary tract symptoms (BPH/LUTS) in the context of systemic metabolic disease. Here we discuss the nature and origins of BPH, examine its role as a component of LUTS and review retrospective clinical studies that have drawn associations between BPH/LUTS and type II diabetes, inflammation and dyslipidemia. PPARgamma signaling, which sits at the nexus of systemic metabolic disease and BPH/LUTS through its regulation of inflammation and insulin resistance, is proposed as a candidate for molecular manipulation in regard to BPH/LUTS. Finally, we introduce new cell and animal models that are being used to study the consequences of obesity, diabetes and inflammation on benign prostatic growth. PMID- 21645963 TI - Treatment of small hepatocellular carcinomas with US-guided high-intensity focused ultrasound. AB - High-intensity focused ultrasound (HIFU) is a noninvasive method that can cause complete coagulation necrosis without requiring the insertion of any instruments. The purpose of this study was to evaluate the safety and efficacy of HIFU treatment for small liver cancers without performing transcatheter arterial chemoembolization (TACE) or rib resection. HIFU ablation was performed without rib resection or the aid of TACE or percutaneous ethanol injection (PEI) in 12 patients with hepatocelullar carcinoma. The HIFU system (Chongqing Haifu Tech, Chongqing, China) was used under ultrasound guidance. All 12 patients completed the treatment without experiencing any adverse events. Complete coagulation was achieved by applying the sonications from the intercostal space when the tumor was located in the right lobe. After treatment, serum alanine aminotransferase (ALT) and serum aspartate aminotransferase (AST) levels were significantly higher than the baseline values; these levels recovered within 1 week. C-reactive protein (CRP) levels increased 1 week after treatment but decreased within 1 month. An epidural anesthetic provided sufficient pain suppression during the procedure. Edema of the subcutaneous tissue was detected in five cases, but the edema disappeared within 1 month. None of the patients developed acute hepatic failure, liver abscess or renal dysfunction. In conclusion, HIFU is effective for the treatment of patients with small liver cancer. PMID- 21645964 TI - High-frequency ultrasound imaging for longitudinal evaluation of non-alcoholic fatty liver disease progression in mice. AB - Nonalcoholic fatty liver disease (NAFLD) is one of the most common causes of hepatic damage in developed countries. For this reason, mouse models of NAFLD have been developed to show progression of the disease because it perfectly resembles the human pathology. Here we show that diagnostic high-frequency ultrasound imaging (US) may be used as an effective method for monitoring the progression of liver disease, from steatosis to hepatocellular carcinoma in the methionine adenosyl transferase and glycine N-methyltransferase-deficient mice models. US reliably detected murine liver lesions associated with NAFLD in the two mice strains tested, with excellent agreement among US images, gross pathology and histological sections. Our results suggest US as a relevant approach for the study of NAFLD in mice, with interesting technical and therapeutic implications. PMID- 21645965 TI - Blood-brain barrier disruption and vascular damage induced by ultrasound bursts combined with microbubbles can be influenced by choice of anesthesia protocol. AB - Numerous animal studies have demonstrated that ultrasound bursts combined with a microbubble-based ultrasound contrast agent can temporarily disrupt the blood brain barrier (BBB) with little or no other apparent effects to the brain. As the BBB is a primary limitation to the use of most drugs in the brain, this method could enable a noninvasive means for targeted drug delivery in the brain. This work investigated whether BBB disruption and vessel damage when overexposure occurs can be influenced by choice of anesthesia protocol, which have different vasoactive effects. Four locations were sonicated transcranially in each brain of 16 rats using an unfocused 532 kHz piston transducer. Burst sonications (10 ms bursts applied at 1 Hz for 60 s) were combined with intravenous Definity (10 MUl/kg) injections. BBB disruption was evaluated using contrast-enhanced MRI. Half of the animals were anesthetized with i.p. ketamine and xylazine, and the other half with inhaled isoflurane and oxygen. Over the range of exposure levels tested, MRI contrast enhancement was significantly higher (p < 0.05) for animals anesthetized with ketamine/xylazine. Furthermore, the threshold for extensive erythrocyte extravasation was lower with ketamine/xylazine. These results suggest that BBB disruption and/or vascular damage can be affected by vascular or other factors that are influenced by different anesthesia protocol. These experiments may also have been influenced by the recently reported findings that the circulation time for perfluorocarbon microbubbles is substantially reduced when oxygen is used as the carrier gas. PMID- 21645966 TI - Acoustic radiation force-driven assessment of myocardial elasticity using the displacement ratio rate (DRR) method. AB - A noninvasive method of characterizing myocardial stiffness could have significant implications in diagnosing cardiac disease. Acoustic radiation force (ARF)-driven techniques have demonstrated their ability to discern elastic properties of soft tissue. For the purpose of myocardial elasticity imaging, a novel ARF-based imaging technique, the displacement ratio rate (DRR) method, was developed to rank the relative stiffnesses of dynamically varying tissue. The basis and performance of this technique was demonstrated through numerical and phantom imaging results. This new method requires a relatively small temporal (<1 ms) and spatial (tenths of mm(2)) sampling window and appears to be independent of applied ARF magnitude. The DRR method was implemented in two in vivo canine studies, during which data were acquired through the full cardiac cycle by imaging directly on the exposed epicardium. These data were then compared with results obtained by acoustic radiation force impulse (ARFI) imaging and shear wave velocimetry, with the latter being used as the gold standard. Through the cardiac cycle, velocimetry results portray a range of shear wave velocities from 0.76-1.97 m/s, with the highest velocities observed during systole and the lowest observed during diastole. If a basic shear wave elasticity model is assumed, such a velocity result would suggest a period of increased stiffness during systole (when compared with diastole). Despite drawbacks of the DRR method (i.e., sensitivity to noise and limited stiffness range), its results predicted a similar cyclic stiffness variation to that offered by velocimetry while being insensitive to variations in applied radiation force. PMID- 21645967 TI - Sunitinib in urothelial cancer: clinical, pharmacokinetic, and immunohistochemical study of predictors of response. AB - BACKGROUND: Sunitinib has activity in patients with metastatic urothelial cancer (UC), but most patients do not respond. OBJECTIVE: To identify predictors of response to sunitinib. DESIGN, SETTING, AND PARTICIPANTS: Seventy-seven patients with advanced UC received sunitinib on one of two schedules at a single institution. Blood pressure (BP), immunohistochemistry (IHC), and pharmacokinetic (PK) results were correlated with response to sunitinib. MEASUREMENTS: BP was assessed on day 1 and 28 of each cycle and on day 14 of cycle 1. IHC was performed on 55 samples from 38 cases using mammalian target of rapamycin and hypoxia-inducible factor (HIF) pathway marker antibodies. Blood samples for PK analysis were collected from 15 patients at three time points. Response was assessed using Response Evaluation Criteria in Solid Tumors criteria. RESULTS AND LIMITATIONS: Sunitinib-induced hypertension predicted improved response when hypertension was categorized as a discrete (p = 0.02) or continuous variable (p = 0.005 [systolic BP] and p = 0.007 [diastolic BP]). The odds ratio of response was 12.5 (95% confidence interval, 1.95-246.8) for grade 3/4 hypertension compared with grade 0. Response was associated with low HIF-1alpha expression in primary (p = 0.07) tissue. A nonstatistically significant trend was seen for an association between greater drug concentration and best response. A correlation between expression markers within the same pathways was identified, phosphorylated-4EBP1 and phosphorylated-S6 (p = 6.5 * 10(-9)), and vascular endothelial growth factor receptor 2 and HIF-1alpha (p = 0.008). Results are limited by small numbers. CONCLUSIONS: Clinical and molecular biomarkers of response to sunitinib may have clinical relevance and require prospective validation. There is an urgent need for predictive biomarkers to guide the management of UC. PMID- 21645968 TI - Removal of reactive red-120 and 4-(2-pyridylazo) resorcinol from aqueous samples by Fe3O4 magnetic nanoparticles using ionic liquid as modifier. AB - The nanoparticles of Fe(3)O(4) as well as the binary nanoparticles of ionic liquid and Fe(3)O(4) (IL-Fe(3)O(4)) were synthesized for removal of reactive red 120 (RR-120) and 4-(2-pyridylazo) resorcinol (PAR) as model azo dyes from aqueous solutions. The mean size and the surface morphology of the nanoparticles were characterized by TEM, DLS, XRD, FTIR and TGA techniques. Adsorption of RR-120 and PAR was studied in a batch reactor at different experimental conditions such as nanoparticle dosage, dye concentration, pH of the solution, ionic strength, and contact time. Experimental results indicated that the IL-Fe(3)O(4) nanoparticles had removed more than 98% of both dyes under the optimum operational conditions of a dosage of 60mg, a pH of 2.5, and a contact time of 2min when initial dyes concentrations of 10-200mg L(-1) were used. The maximum adsorption capacity of IL Fe(3)O(4) was 166.67 and 49.26mg g(-1) for RR-120 and PAR, respectively. The isotherm experiments revealed that the Langmuir model attained better fits to the equilibrium data than the Freundlich model. The Langmuir adsorption constants were 5.99 and 3.62L mg(-1) for adsorptions of RR-120 and PAR, respectively. Both adsorption processes were endothermic and dyes could be desorbed from IL Fe(3)O(4) by using a mixed NaCl-acetone solution and adsorbent was reusable. PMID- 21645969 TI - Kinetics of 2,6-dimethylaniline oxidation by various Fenton processes. AB - The kinetics of 2,6-dimethylaniline degradation by Fenton process, electro-Fenton process and photoelectro-Fenton process was investigated. This study attempted to eliminate the potential interferences from intermediates by making a kinetics comparison of Fenton, electro-Fenton and photoelectro-Fenton methods through use initial rate techniques during the first 10 min of the reaction. Exactly how the initial concentration of 2,6-dimethylaniline, ferrous ions and hydrogen peroxide affects 2,6-dimethylaniline degradation was also examined. Experimental results indicate that the 2,6-dimethylaniline degradation in the photoelectro-Fenton process is superior to the ordinary Fenton and electro-Fenton processes. Additionally, for 100% removal of 1mM 2,6-dimethylaniline, the supplementation of 1mM of ferrous ion, 20mM of hydrogen peroxide, current density at 15.89 A m(-2) and 12 UVA lamps at pH 2 was necessary. The overall rate equations for 2,6 dimethylaniline degradation by Fenton, electro-Fenton and photoelectro-Fenton processes were proposed as well. PMID- 21645970 TI - Relating organic fouling of reverse osmosis membranes to adsorption during the reclamation of secondary effluents containing methylene blue and rhodamine B. AB - Dyes fouling of reverse osmosis (RO) membranes and its relation to adsorption had been investigated by using a crossflow RO filtration setup. Methylene blue (MB) and rhodamine B (RB) were used as model organic foulants. The calculated amount of the irreversible sorption was related to the irreversible flux decline. The characteristic fouling kinetics was accounted by Langmuir-Hinshelwood (L-H) kinetics model for initial fouling, with the fouling rate constant k=0.0556MUm s( 1)min(-1) and k=0.0181MUm s(-1)min(-1) for MB and RB fouling RO membrane CPA2, respectively. And the subsequent fouling was attributed to the growth of a dye cake. A remarkable correlation was obtained between the quantified irreversible sorption and irreversible flux decline under the solution chemistries investigated. In the presence of divalent cation, the extent of flux decline was related to the competition model. PMID- 21645971 TI - Cystic echinococcosis in Sardinia: farmers' knowledge and dog infection in sheep farms. AB - Cystic Echinococcosis (CE) is one of the most widespread parasitic diseases in Sardinia, the second largest Mediterranean island where almost 3,558,000 milk sheep were raised extensively. The aim of this survey was to evaluate the level of farmers' knowledge on CE transmission, focusing on the role of human to facilitate the persistence of this zoonosis in Sardinia after 14 years after the last campaign against CE. The other goal of the survey is to update on presence of Echinococcus granulosus in its definitive hosts through three ELISA coproantigen tests. An interview was carried out with 172 farmers. The questionnaire was designed to include possible factors associated with the transmission of Echinococcosis: ownership and number of dogs, the use of anthelmintic drugs against dog cestode, frequency of anthelmintic treatment in dogs, home slaughtering and offal disposal. Individual faecal samples were retrieved from 300 dogs, and after a preliminary macroscopic examination to discover adult worms and/or proglottids, was submitted to copromicroscopic examination. Coproantigens were then extracted according to the protocol described by Allan et al. (1992), and subsequently stored at -20 degrees C until use. Faecal soluble antigens from E. granulosus were detected using three different ELISA coproantigen assays: (a) the commercially produced Chekit Echinotest (Bommeli, Bern, CH) based on polyclonal antibodies against adult excretory/secretory (E/S) antigens; (b) a sandwich ELISA that uses rabbit polyclonal antibodies against adult E/S antigens and biotinylated monoclonal antibody EmA9 produced against adult Echinococcus multilocularis somatic extract (Malgor et al., 1997); and (c) a sandwich assay that uses monoclonal antibody EgC3 produced by immunization with adult E. granulosus E/S products (Casaravilla et al., 2005). Questionnaire results reveal that on all farms home-slaughtering was done, and offal was used as dog meal raw (17%) or after boiling (37%), discarded in the trash (23%), or buried superficially (15%). Most farmers (69%) declared to deworm their dogs, but only 10% used cestodicidal drugs. The coprological survey of 300 farm dogs using sedimentation, flotation and three different coproantigen (CA) ELISAs resulted in a faecal prevalence of 8.3% for taeniid eggs, while the CA tests gave prevalences of 3% (Chekit Echinotest, Bommeli), 6% (EmA9 sandwich ELISA) and 10% (EgC3 sandwich ELISA). Our results show that this is not only an educational problem, but also an economic one, stressing the need that future control plans should follow an integrative approach including veterinary and medical services, farmers, breeders' associations and the Government. PMID- 21645972 TI - Immune modulatory agents in hematopoietic malignancies. AB - Immune modulatory drugs have been successfully used to treat patients with multiple myeloma (MM), myelodysplastic syndromes displaying loss of 5q (del5q MDS) and chronic lymphocytic leukemia (CLL). Immune modulatory drugs are used in first-line therapy in combination with functionally complementary compounds, but have also shown efficacy in refractory disease. However, their exact mode of action remains unclear. Here we describe the clinical impact of these compounds on MM, del5q MDS and CLL, discuss their mode of action with respect to intracellular targets, focus on the phenotypic changes that immune modulatory compounds induce in the tumor microenvironment and how they modulate the immune response. PMID- 21645973 TI - Glutamatergic hyperfunctioning during alcohol withdrawal syndrome: therapeutic perspective with zinc and magnesium. AB - It is known that the glutamatergic pathways are hyperfunctioning during alcohol withdrawal syndrome. It has been demonstrated that hyperfunctioning of this system causes a great damage to the superior cortical activity, the ability to concentrate and the control of impulses. Recent studies show that the cations zinc and magnesium modulate the glutamatergic function, reducing it to non-toxic levels, yet not reducing it to the point of depriving this neurotransmitter of its normal activity. New perspectives of treatment focus on the modulation of this system, having, as a result, reestablishment of impulse control abilities, damage prevention to the hippocampus and the amygdala and prevention of future relapses. PMID- 21645974 TI - The effects of feeding flaxseed to beef cows given forage based diets on fatty acids of longissimus thoracis muscle and backfat. AB - This study was conducted to investigate changes in fatty acid profiles of beef cows fed grass hay or barley silage based diets, with or without flaxseed supplementation. Both flaxseed and hay feeding increased levels of alpha linolenic acid (LNA; 18:3n-3) in longissimus thoracis and backfat (P<0.001). A forage type by flaxseed level interaction was observed for most LNA biohydrogenation intermediates (P<0.05) that indicated feeding hay combined with flaxseed led to the greatest levels of total conjugated linolenic acid, total conjugated linoleic acid, total non-conjugated dienes and total trans-18:1. Predominant biohydrogenation intermediates included t11,c15-18:2, rumenic acid (c9,t11-18:2) and vaccenic acid (t11-18:1). PMID- 21645975 TI - Physico-chemical and sensory properties of reduced-fat mortadella prepared with blends of calcium, magnesium and potassium chloride as partial substitutes for sodium chloride. AB - Blends of calcium, magnesium and potassium chloride were used to partially replace sodium chloride (50-75%) in reduced-fat mortadella formulations. The presence of calcium chloride reduced the emulsion stability, cooking yield, elasticity and cohesiveness and increased hardness; however, it yielded the best sensory acceptance when 50% NaCl was replaced by 25% CaCl(2) and 25% KCl. There was no effect of the salt substitutes on mortadella color, appearance and aroma. All salt combinations studied showed stable lipid oxidation during its shelf life. The use of a blend with 1% NaCl, 0.5% KCl and 0.5% MgCl(2) resulted in the best emulsion stability, but the worst scores for flavor. This study suggests that it is possible to reduce the sodium chloride concentration by 50% in reduced fat mortadella using the studied salt combinations with necessary adjustments to optimize the sensory properties (MgCl(2) 25%; KCl 25%) or emulsion stability (CaCl(2) 25%; KCl 25%). PMID- 21645976 TI - Continued benefit to androgen deprivation therapy for prostate cancer patients treated with dose-escalated radiation therapy across multiple definitions of high risk disease. AB - PURPOSE: To analyze prognostic factors in patients with high-risk prostate cancer treated with dose-escalated external-beam radiation therapy (EBRT) and androgen deprivation (ADT). METHODS AND MATERIALS: Between 1998 and 2008 at the University of Michigan Medical Center, 718 men were consecutively treated with EBRT to at least 75 Gy. Seven definitions of high-risk prostate cancer, applying to 11-33% of patients, were evaluated. Biochemical failure (BF), salvage ADT use, metastatic progression, and prostate cancer-specific mortality (PCSM) were estimated by the Kaplan-Meier method and Cox proportional hazards regression. RESULTS: Each high-risk definition was associated with increased BF (hazard ratio [HR] 2.8-3.9, p < 0.0001), salvage ADT use (HR 3.9-6.3, p < 0.0001), metastasis (HR 3.7-6.6, p < 0.0001), and PCSM (HR 3.7-16.2, p < 0.0001). Furthermore, an increasing number of high-risk features predicted worse outcome. Adjuvant ADT yielded significant reductions in both metastases (HR 0.19-0.38, p < 0.001) and PCSM (HR 0.38-0.50, p < 0.05) for all high-risk definitions (with the exception of clinical Stage T3-4 disease) but improved BF only for those with elevated Gleason scores (p < 0.03, HR 0.25-0.48). When treated with ADT and dose-escalated EBRT, patients with Gleason scores 8 to 10, without other high-risk features, had 8-year freedom from BF of 74%, freedom from distant metastases of 93%, and cause specific survival of 92%, with salvage ADT used in 16% of patients. CONCLUSION: Adjuvant ADT results in a significant improvement in clinical progression and PCSM across multiple definitions of high-risk disease even with dose-escalated EBRT. There is a subset of patients, characterized by multiple high-risk features or the presence of Gleason Pattern 5, who remain at significant risk for metastasis and PCSM despite current treatment. PMID- 21645977 TI - Stereotactic body radiotherapy for primary hepatocellular carcinoma. AB - PURPOSE: To evaluate the safety and efficacy of stereotactic body radiotherapy (SBRT) for the treatment of primary hepatocellular carcinoma (HCC). METHODS AND MATERIALS: From 2005 to 2009, 60 patients with liver-confined HCC were treated with SBRT at the Indiana University Simon Cancer Center: 36 Child-Turcotte-Pugh (CTP) Class A and 24 CTP Class B. The median number of fractions, dose per fraction, and total dose, was 3, 14 Gy, and 44 Gy, respectively, for those with CTP Class A cirrhosis and 5, 8 Gy, and 40 Gy, respectively, for those with CTP Class B. Treatment was delivered via 6 to 12 beams and in nearly all cases was prescribed to the 80% isodose line. The records of all patients were reviewed, and treatment response was scored according to Response Evaluation Criteria in Solid Tumors v1.1. Toxicity was graded according to the Common Terminology Criteria for Adverse Events v4.0. Local control (LC), time to progression (TTP), progression-free survival (PFS), and overall survival (OS) were calculated according to the method of Kaplan and Meier. RESULTS: The median follow-up time was 27 months, and the median tumor diameter was 3.2 cm. The 2-year LC, PFS, and OS were 90%, 48%, and 67%, respectively, with median TTP of 47.8 months. Subsequently, 23 patients underwent transplant, with a median time to transplant of 7 months. There were no >=Grade 3 nonhematologic toxicities. Thirteen percent of patients experienced an increase in hematologic/hepatic dysfunction greater than 1 grade, and 20% experienced progression in CTP class within 3 months of treatment. CONCLUSIONS: SBRT is a safe, effective, noninvasive option for patients with HCC <=6 cm. As such, SBRT should be considered when bridging to transplant or as definitive therapy for those ineligible for transplant. PMID- 21645978 TI - The role of RTX toxins in host specificity of animal pathogenic Pasteurellaceae. AB - RTX toxins are bacterial pore-forming toxins that are particularly abundant among pathogenic species of Pasteurellaceae, in which they play a major role in virulence. RTX toxins of several primary pathogens of the family of Pasteurellaceae are directly involved in causing necrotic lesions in the target organs. Many RTX toxins are known as haemolysins because they lyse erythrocytes in vitro, an effect that is non-specific, but which serves as a useful marker in bacteriological identification and as an easily measurable signal in vitro in experimental studies. More recent studies have shown that the specific targets of most RTX toxins are leukocytes, with RTX toxins binding to the corresponding beta subunit (CD18) of beta2 integrins and then exerting cytotoxic activity. After uptake by the target cell, at sub-lytic concentrations, some RTX toxins are transported to mitochondria and induce apoptosis. For several RTX toxins the binding to CD18 has been shown to be host specific and this seems to be the basis for the host range specificity of these RTX toxins. Observations on two very closely related species of the Pasteurellaceae family, Actinobacillus suis, a porcine pathogen particularly affecting suckling pigs, and Actinobacillus equuli subsp. haemolytica, which causes pyosepticaemia in new-born foals (sleepy foal disease), have revealed that they express different RTX toxins, named ApxI/II and Aqx, respectively. These RTX toxins are specifically cytotoxic for porcine and equine leukocytes, respectively. Furthermore, the ApxI and Aqx toxins of these species, when expressed in an isogenetic background in Escherichia coli, are specifically cytotoxic for leukocytes of their respective hosts. These data indicate the determinative role of RTX toxins in host specificity of pathogenic species of Pasteurellaceae. PMID- 21645979 TI - Hair testing and self-report of cocaine use. AB - Hair analysis is a useful tool in both clinical and forensic fields: it allows information about drugs of abuse (DOA) consumption to be obtained. However, in spite of analytical results, sometimes patients continue to deny using drugs or, on the contrary, insist on describing themselves as severe drug addicts; indeed there are often considerable difficulties in getting truthful statements about the real amount of drugs used. In this study we have tried to compare cocaine concentration in hair samples with self-reported drug intake. We enrolled 113 subjects (61 Africans, 52 Caucasians) who had been recently sent to jail. They were asked to tell about their use of illicit drugs during the last three months and then submitted to hair analysis. Hair segments (3 cm) were analyzed by GC-MS for amphetamines, cocaine and opiates. Useful data was obtained from 82 subjects, separated into two main groups on account of ethnic origin (African or Caucasian) and divided further into daily, weekly and monthly users. The results showed qualitative results and self-reported consumption to be in good agreement, although the correlation between frequency of consumption and concentration in hair revealed sometimes higher concentrations in contrast with the admission of low consumption. There was a definite separation between occasional and daily use (especially in Caucasian people), while concentrations found where weekly use was reported were more variable. Concentrations of cocaine measured in Africans' hair were much higher than in Caucasians'. Even if this study is exclusively based on self-report, it provides some interesting information in order to differentiate the frequency of consumption, and especially underlines the great importance of ethnic bias on hair analysis. PMID- 21645980 TI - Clinical investigation on application of water swallowing to MR esophagography. AB - OBJECTIVE: To verify the clinical outcomes of applying water swallowing to MR esophagography. METHODS: Thirty patients confirmed postoperatively or histopathologically with thoracic esophageal carcinoma by endoscopic biopsy and 10 healthy volunteers with normal esophagus underwent respectively conventional magnetic resonance imaging (MRI) detection and water swallowing MR esophagography. Of those patients, 4 underwent second examination after radiotherapy. Assessment on imaging effects of MR esophagography was performed. Assessment on definition on MR esophagography of the tumor in both upper and lower ends, specific localization, tumor size finally measured, coincidence with the gross pathologic types and tumor staging were respectively performed by comparison with conventional MRI. Additionally, we evaluated the outcomes of radiotherapy by comparing the previous MR esophagography with the second one with interventional technique. RESULTS: Of the total 44 images of MR esophagography, 97.7% (43/44) were in high resolution by sagittal view and 81.8% (36/44) by cross section. 93.3% (56/60) of the MR esophagography were clearly defined with the neoplastic lesion ends in the 30 patients with thoracic esophageal carcinoma, compared with 11.7% (7/60) by conventional MRI. The results were totally different in statistics (P<0.005). Preoperative conventional MRI detection of the 22 cases in 25 undergone radical resection suggested vague diameter of the primary tumor and impossibly identified it at middle-lower thoracic esophagus in 5, and even failed to confirm gross pathologic types in 19 cases. Yet, MR esophagography with water swallowing represented accurate tumor length (graded as excellent) in 88% (22/25), localization in 100% (25/25), exact gross pathologic types in 88% (22/25), and accuracy for tumor staging in 80.8% (21/26) compared to 92.3% (24/26) by conventional MRI. Therapeutic effects achieved in 4 patients with radiotherapy. CONCLUSIONS: MR esophagography with water swallowing makes optimal esophagram and is of great value in the preoperative diagnosis of thoracic esophageal cancer and assessment of the radiotherapy effects for patients with such neoplasm, which may serve as an alternative for conventional MRI. PMID- 21645981 TI - Estimated effective dose of CT-guided percutaneous cryoablation of liver tumors. AB - PURPOSE: To estimate effective dose during CT-guided cryoablation of liver tumors, and to assess which procedural factors contribute most to dose. MATERIALS AND METHODS: Our institutional review board approved this retrospective, HIPAA compliant study. A total of 20 CT-guided percutaneous liver tumor cryoablation procedures were performed in 18 patients. Effective dose was determined by multiplying the dose length product for each CT scan obtained during the procedure by a conversion factor (0.015mSv/mGy-cm), and calculating the sum for each phase of the procedure: planning, targeting, monitoring, and post-ablation survey. Effective dose of each phase was compared using a repeated measures analysis. Using Spearman correlation coefficients, effective doses were correlated with procedural factors including number of scans, ratio of targeting distance to tumor size, anesthesia type, number of applicators, performance of ancillary procedures (hydrodissection and biopsy), and use of CT fluoroscopy. RESULTS: Effective dose per procedure was 72+/-18mSv. The effective dose of targeting (37.5+/-12.5mSv) was the largest component compared to the effective dose of the planning phase (4.8+/-2.2mSv), the monitoring phase (25.5+/-6.8mSv), and the post-ablation survey (4.1+/-1.9mSv) phase (p<0.05). Effective dose correlated positively only with the number of scans (p<0.01). CONCLUSIONS: The effective dose of CT-guided percutaneous cryoablation of liver tumors can be substantial. Reducing the number of scans during the procedure is likely to have the greatest effect on lowering dose. PMID- 21645982 TI - Detection of Crohn's disease: Comparison of CT and MR enterography without anti peristaltic agents performed on the same day. AB - OBJECTIVE: To directly compare CT enterography (CTE) and MR enterography (MRE) without antiperistaltic agents. MATERIALS/METHODS: 26 patients referred for CTE underwent CTE immediately followed by MRE without use of an anti-peristaltic agent. Each study was evaluated on a 10 point scale for exam quality, level of diagnostic confidence, and presence of Crohn's disease. Kappa analysis was performed to determine the degree of agreement between the CTE and MRE of each patient. RESULTS: 25 patients completed the MRE. The quality of the CTEs was judged as excellent by both readers (reader 1=average 9.5/10, reader 2=average 9.1/10). The quality of the MREs was ranked lower than the CTEs by both readers (reader 1=average 8.9/10, reader 2=average 7.2/10), which was statistically significant (p<0.05). The level of confidence in interpretation was not significantly different between CTE and MRE for reader 1 or 2 (p=0.3). There was substantial agreement between readers for the presence or absence of Crohn's disease on both CTE (kappa=0.75) and MRE (kappa=0.67). CONCLUSION: MR enterography without anti-peristaltic agents results in high diagnostic confidence and excellent agreement for the presence of Crohn's disease. PMID- 21645983 TI - Geobacillus galactosidasius sp. nov., a new thermophilic galactosidase-producing bacterium isolated from compost. AB - Two thermophilic spore-forming strains, with optimum growth temperature at 70 degrees C, were isolated from compost of the "Experimental System of Composting" (Teora, Avellino, Italy). A phylogenetic analysis based on 16S rRNA gene sequences showed that these organisms represented a new species of the genus Geobacillus. Based on polyphasic taxonomic data the strains represented a novel species for which the name Geobacillus galactosidasius sp. nov. is proposed. The type strain is CF1B(T) (=ATCC BAA-1450(T)=DSM 18751(T)). PMID- 21645984 TI - Penetrating atherosclerotic ulcerative disease of the aorta: do emergency physicians need to worry? AB - BACKGROUND: In recent years, different aortic pathologies have been grouped together and described in terms of a spectrum of disease referred to as Acute Aortic Syndrome (AAS). Overlapping of these conditions has traditionally introduced discrepancy to the understanding of the pathophysiology and definitive care. Penetrating atherosclerotic ulcer (PAU), an ulceration of an atherosclerotic plaque, has been increasingly recognized as the individual cause of the acute aortic pathology. OBJECTIVE: The natural evolution and complications of PAU are described, as well as the current diagnostic strategies, definitive management options, and initial care in the emergency department. DISCUSSION: The natural history of PAU is variable but is generally considered to be an AAS with a high incidence of complications. The clinical differentiation of symptomatic PAU from other causes of AAS is very challenging. Computed tomography imaging has provided additional accuracy with the use of the retrospective electrocardiographic gating technique. Despite traditional surgical therapies for aortic pathology, later studies have demonstrated the success of non-operative management, such as endoluminal grafting. CONCLUSION: The relative rarity of this disease, the difficulties in its detection, and the lack of precise guidelines about its management may result in diagnostic delays. Asymptomatic patients with well-controlled blood pressure are safely managed as outpatients in close consultation with vascular surgery. PMID- 21645985 TI - Factors influencing administration of hepatitis B vaccine to community-dwelling teenagers aged 12-18 with an intellectual disability. AB - The study aims to determine hepatitis B vaccination coverage rates among community-dwelling teenagers with an intellectual disability in Taiwan and to identify the possible influencing factors of their vaccination. The present paper was part of the results of the "2007 National Survey on Healthy Behaviors and Preventive Health Utilizations of People with Intellectual Disabilities in Taiwan," which was a cross-sectional survey of 1111 intellectually disabled (ID) teenagers ranging from 12 to 18 years of age. The results showed that the completed hepatitis B vaccination rate was 72.9%, a rate lower than that in the general population of Taiwan considering the same age group. There was no gender difference between each age group in the vaccination rate in this population. Multilevel logistic regression analyses revealed that those ID individuals whose primary caregivers were parents or siblings (OR = 2.45, 95% CI = 1.29-4.64), whose household monthly income was 20,000-59,999 NTD vs. less than 20,000 NTD (OR = 2.47, 95% CI = 1.00-6.12), and who had ever undergone an oral health exam (OR = 2.29, 95% CI = 1.24-4.01) were more likely to receive a complete hepatitis B vaccination than their counterparts. The study highlighted that most teenagers had received complete hepatitis B vaccination. Nonetheless, better public health strategies may be needed to deliver the hepatitis B vaccine to those who do not comply with the vaccination schedule in the community. PMID- 21645986 TI - Reduced sensitivity to slow-rate dynamic auditory information in children with dyslexia. AB - The etiology of developmental dyslexia remains widely debated. An appealing theory postulates that the reading and spelling problems in individuals with dyslexia originate from reduced sensitivity to slow-rate dynamic auditory cues. This low-level auditory deficit is thought to provoke a cascade of effects, including inaccurate speech perception and eventually unspecified phoneme representations. The present study investigated sensitivity to frequency modulation and amplitude rise time, speech-in-noise perception and phonological awareness in 11-year-old children with dyslexia and a matched normal-reading control children. Group comparisons demonstrated that children with dyslexia were less sensitive than normal-reading children to slow-rate dynamic auditory processing, speech-in-noise perception, phonological awareness and literacy abilities. Correlations were found between slow-rate dynamic auditory processing and phonological awareness, and speech-in-noise perception and reading. Yet, no significant correlation between slow-rate dynamic auditory processing and speech in-noise perception was obtained. Together, these results indicate that children with dyslexia have difficulties with slow-rate dynamic auditory processing and speech-in-noise perception and that these problems persist until sixth grade. PMID- 21645987 TI - Assisting people with multiple disabilities and minimal motor behavior to improve computer Drag-and-Drop efficiency through a mouse wheel. AB - This study evaluated whether two people with multiple disabilities and minimal motor behavior would be able to improve their Drag-and-Drop (DnD) performance using their finger/thumb poke ability with a mouse scroll wheel through a Dynamic Drag-and-Drop Assistive Program (DDnDAP). A multiple probe design across participants was used in this study to assess the effects of using DDnDAP in enhancing participants' DnD ability. Both participants: (a) improved their DnD efficiency with the use of DDnDAP and (b) remained highly successful through the maintenance phase. The implications of the findings are discussed. PMID- 21645988 TI - Use of activity schedule to promote independent performance of individuals with autism and other intellectual disabilities: a review. AB - A literature review was conducted on the effectiveness of activity schedules. Twenty three studies that a) were peer-reviewed, b) were experimental, c) implemented activity schedule as a primary intervention, d) incorporated multiple activities, and e) aimed to teach learners to self-manage individual schedules were included in the review. The results demonstrated the effectiveness of activity schedules for promoting independence and self-management skills for a broad range of individuals with intellectual disabilities. An increase in engagement and on-task behavior was the most frequently cited outcome, followed by independent task initiation or transition and self-scheduling. Failure to include social validity measures and caregivers as interventionists were discussed. A corpus of the reviewed studies supports applications of activity schedule in school and (group) home settings. PMID- 21645989 TI - Teaching students with developmental disabilities to operate an iPod Touch((r)) to listen to music. AB - We evaluated an intervention procedure for teaching three students with developmental disabilities to independently operate a portable multimedia device (i.e., an iPod Touch((r))) to listen to music. The intervention procedure included the use of video modeling, which was presented on the same iPod Touch((r)) that the students were taught to operate to listen to music. Four phases (i.e., baseline, intervention, fading, and follow-up) were arranged in accordance with a delayed multiple-probe across participants design. During baseline, the students performed from 25 to 62.5% of the task analyzed steps correctly. With intervention, all three students correctly performed 80-100% of the steps and maintained this level of performance when video modeling was removed and during follow-up. The findings suggest that the video modeling procedure was effective for teaching the students to independently operate a portable multimedia device to access age-appropriate leisure content. PMID- 21645990 TI - Numerical magnitude processing in children with mild intellectual disabilities. AB - The present study investigated numerical magnitude processing in children with mild intellectual disabilities (MID) and examined whether these children have difficulties in the ability to represent numerical magnitudes and/or difficulties in the ability to access numerical magnitudes from formal symbols. We compared the performance of 26 children with MID on a symbolic (digits) and a non-symbolic (dot-arrays) comparison task with the performance of two control groups of typically developing children: one group matched on chronological age and one group matched on mathematical ability level. Findings revealed that children with MID performed more poorly than their typically developing chronological age matched peers on both the symbolic and non-symbolic comparison tasks, while their performance did not substantially differ from the ability-matched control group. These findings suggest that the development of numerical magnitude representation in children with MID is marked by a delay. This performance pattern was observed for both symbolic and non-symbolic comparison tasks, although difficulties on the former task were more prominent. Interventions in children with MID should therefore foster both the development of magnitude representations and the connections between symbols and the magnitudes they represent. PMID- 21645991 TI - Reproducibility and inter-vendor variability of left ventricular deformation measurements by three-dimensional speckle-tracking echocardiography. AB - BACKGROUND: Myocardial deformation measurements using two-dimensional speckle tracking echocardiography (STE) are known to vary among vendors. The intervendor agreement of three-dimensional (3D) deformation indices has not been studied. The goals of this study were to determine the intervendor agreement of 3D STE-based measurements of left ventricular (LV) deformation parameters to investigate the intrinsic variability of these measurements and identify the sources of intervendor differences. METHODS: Real-time full-volume images obtained in 30 subjects with normal LV systolic function using two vendors' equipment (V1 and V2) on the same day were analyzed by two independent observers using two software packages (S1 and S2). Agreement between three technique combinations (V1/S1, V2/S2, and V1/S2) and their intrinsic reproducibility (interobserver and intraobserver agreement) were assessed using intraclass correlation coefficients. Parameters of LV deformation included global longitudinal strain, twist, 3D displacement, and 3D strain and its radial, longitudinal, and circumferential components. RESULTS: For all three combinations, intertechnique agreement was poor (intraclass correlation coefficient < 0.4), always beyond the intrinsic variability. For all measured parameters, the intertechnique agreement was better when the same software package was used with images from different vendors (V2/S2 vs V1/S2) than when images from same vendor were analyzed using different software (V1/S2 vs V1/S1). CONCLUSIONS: Three-dimensional STE-derived LV deformation parameters are highly vendor dependent, and the discordance levels are beyond intrinsic measurement variability of any of the tested combinations of imaging equipment and analysis software. This intervendor discordance must be taken into account when interpreting 3D deformation data. PMID- 21645992 TI - Feasibility of single-beat full-volume capture real-time three-dimensional echocardiography and auto-contouring algorithm for quantification of left ventricular volume: validation with cardiac magnetic resonance imaging. AB - BACKGROUND: With recent developments in echocardiographic technology, a new system using real-time three-dimensional echocardiography (RT3DE) that allows single-beat acquisition of the entire volume of the left ventricle and incorporates algorithms for automated border detection has been introduced. Provided that these techniques are acceptably reliable, three-dimensional echocardiography may be much more useful for clinical practice. The aim of this study was to evaluate the feasibility and accuracy of left ventricular (LV) volume measurements by RT3DE using the single-beat full-volume capture technique. METHODS: One hundred nine consecutive patients scheduled for cardiac magnetic resonance imaging and RT3DE using the single-beat full-volume capture technique on the same day were recruited. LV end-systolic volume, end-diastolic volume, and ejection fraction were measured using an auto-contouring algorithm from data acquired on RT3DE. The data were compared with the same measurements obtained using cardiac magnetic resonance imaging. RESULTS: Volume measurements on RT3DE with single-beat full-volume capture were feasible in 84% of patients. Both interobserver and intraobserver variability of three-dimensional measurements of end-systolic and end-diastolic volumes showed excellent agreement. Pearson's correlation analysis showed a close correlation of end-systolic and end-diastolic volumes between RT3DE and cardiac magnetic resonance imaging (r = 0.94 and r = 0.91, respectively, P < .0001 for both). Bland-Altman analysis showed reasonable limits of agreement. After application of the auto-contouring algorithm, the rate of successful auto-contouring (cases requiring minimal manual corrections) was <50%. CONCLUSIONS: RT3DE using single-beat full-volume capture is an easy and reliable technique to assess LV volume and systolic function in clinical practice. However, the image quality and low frame rate still limit its application for dilated left ventricles, and the automated volume analysis program needs more development to make it clinically efficacious. PMID- 21645993 TI - Shrinkage based assessment of surgical margins: a need of the hour in oral cancer. PMID- 21645995 TI - Focal epilepsy of probable temporal lobe origin in a Gypsy family showing linkage to a novel locus on 7p21.3. AB - We aimed to characterise the phenotype and perform genetic studies in a family of Roma/Gypsy ethnicity, affected by epilepsy. The mean age at onset of epilepsy was 9 years and seizures persisted into adulthood. Antecedent febrile convulsions were rare. Seizure semiology and EEG findings suggested mesial temporal lobe origin with no evidence of hippocampal sclerosis. Seizures frequently generalised. Family structure suggested autosomal-dominant inheritance with incomplete penetrance. Linkage analysis identified a single novel locus on 7p21.3, corresponding to the expected maximum in the family. Previously reported temporal lobe epilepsy (TLE) loci were definitely excluded. The minimal shared haplotype of 2.4cM (1.3Mb) was not observed in other affected families or controls from the same population. Three brain-expressed validated genes in the critical region represent potential candidates. We have identified an epilepsy syndrome with temporal lobe seizures commonly evolving to generalised convulsions. Linkage to 7p21.3 adds up to a total of five currently known FTLE loci. PMID- 21645996 TI - Evidence for association of hyperprolinemia with schizophrenia and a measure of clinical outcome. AB - There are multiple genetic links between schizophrenia and a deficit of proline dehydrogenase (PRODH) enzyme activity. However, reports testing for an association of schizophrenia with the resulting proline elevation have been conflicting. The objectives of this study were to investigate whether hyperprolinemia is associated with schizophrenia, and to measure the relationship between plasma proline, and clinical features and symptoms of schizophrenia. We performed a cross-sectional case-control study, comparing fasting plasma proline in 90 control subjects and 64 schizophrenic patients and testing for association of mild to moderate hyperprolinemia with schizophrenia. As secondary analyses, the relationship between hyperprolinemia and five measures of clinical onset, symptoms and outcome were investigated. Patients had significantly higher plasma proline than matched controls (p<0.0001), and categorical analysis of gender adjusted hyperprolinemia showed a significant association with schizophrenia (OR 6.15, p=0.0003). Hyperprolinemic patients were significantly older at their first hospitalization (p=0.015 following correction for multiple testing). While plasma proline level was not related to total, positive or negative symptoms, hyperprolinemic status had a significant effect on length of hospital stay (p=0.005), following adjustment for race, BPRS score, and cross-sectional time from admission to proline measurement. Mild to moderate hyperprolinemia is a significant risk factor for schizophrenia, and may represent an intermediate phenotype in the disease. Hyperprolinemic patients have a significantly later age of first psychiatric hospitalization, suggestive of later onset, and hospital stays 46% longer than non-hyperprolinemic subjects. These findings have implications in the etiology of schizophrenia, and for the clinical management of these patients. PMID- 21645997 TI - A broad cortical reserve accelerates response to cognitive enhancement therapy in early course schizophrenia. AB - OBJECTIVES: Cognitive rehabilitation can improve cognition in schizophrenia and prevent disability. It is unknown, however, whether a greater neurobiologic reserve, as measured by cortical volumes, will predict a favorable response to rehabilitation. We investigated this question in early course schizophrenia patients treated with Cognitive Enhancement Therapy (CET). METHODS: Outpatients in the early course of schizophrenia or schizoaffective disorder were randomly assigned to CET (n=29) or an Enriched Supportive Therapy control (n=21) and treated for two years. Cortical surface area and gray matter volume data were collected before treatment using structural magnetic resonance imaging. Neurocognition and social cognition were assessed before, and after one and two years of treatment. Moderator analyses examined the impact of pre-treatment cortical surface area and gray matter volume on differential neurocognitive and social-cognitive response to CET. RESULTS: Pre-treatment, whole brain cortical surface area and gray matter volume significantly moderated the effects of CET on social cognition, but not neurocognition. Greater neurobiologic reserve predicted a rapid social-cognitive response to CET in the first year of treatment; patients with less neurobiologic reserve achieved a comparable social-cognitive response by the second year. While nearly every regional measurement significantly contributed to this accelerated social-cognitive treatment response, effects were the strongest in the temporal cortex. CONCLUSIONS: A broad cortical surface area and gray matter reserve is associated with an accelerated social-cognitive response to CET in early schizophrenia, yet the benefits of cognitive rehabilitation are achieved in those with less initial cognitive resources after a longer duration of treatment. PMID- 21645999 TI - Artificial Intelligence in Medicine AIME 2009. PMID- 21645998 TI - Are we studying and treating schizophrenia correctly? AB - New findings are rapidly revealing an increasingly detailed image of neural- and molecular-level dysfunction in schizophrenia, distributed throughout interconnected cortico-striato-pallido-thalamic circuitry. Some disturbances appear to reflect failures of early brain maturation, that become codified into dysfunctional circuit properties, resulting in a substantial loss of, or failure to develop, both cells and/or appropriate connectivity across widely dispersed brain regions. These circuit disturbances are variable across individuals with schizophrenia, perhaps reflecting the interaction of multiple different risk genes and epigenetic events. Given these complex and variable hard-wired circuit disturbances, it is worth considering how new and emerging findings can be integrated into actionable treatment models. This paper suggests that future efforts towards developing more effective therapeutic approaches for the schizophrenias should diverge from prevailing models in genetics and molecular neuroscience, and focus instead on a more practical three-part treatment strategy: 1) systematic rehabilitative psychotherapies designed to engage healthy neural systems to compensate for and replace dysfunctional higher circuit elements, used in concert with 2) medications that specifically target cognitive mechanisms engaged by these rehabilitative psychotherapies, and 3) antipsychotic medications that target nodal or convergent circuit points within the limbic motor interface, to constrain the scope and severity of psychotic exacerbations and thereby facilitate engagement in cognitive rehabilitation. The use of targeted cognitive rehabilitative psychotherapy plus synergistic medication has both common sense and time-tested efficacy with numerous other neuropsychiatric disorders. PMID- 21646000 TI - Modern parameterization and explanation techniques in diagnostic decision support system: a case study in diagnostics of coronary artery disease. AB - OBJECTIVE: Coronary artery disease has been described as one of the curses of the western world, as it is one of its most important causes of mortality. Therefore, clinicians seek to improve diagnostic procedures, especially those that allow them to reach reliable early diagnoses. In the clinical setting, coronary artery disease diagnostics are typically performed in a sequential manner. The four diagnostic levels consist of evaluation of (1) signs and symptoms of the disease and electrocardiogram at rest, (2) sequential electrocardiogram testing during the controlled exercise, (3) myocardial perfusion scintigraphy, and (4) finally coronary angiography, that is considered as the "gold standard" reference method. Our study focuses on improving diagnostic performance of the third, virtually non invasive, diagnostic level. METHODS AND MATERIALS: Myocardial scintigraphy results in a series of medical images that are obtained by relatively inexpensive means. In clinical practice, these images are manually described (parameterized) by expert physicians. In the paper we present an innovative alternative to manual image evaluation-an automatic image parameterization on multiple resolutions, based on texture description with specialized association rules. Extracted image parameters are combined into more informative composite parameters by means of principal component analysis, and finally used to build automatic classifiers with machine learning methods. RESULTS: Our experiments with synthetic datasets show that association-rule-based multi-resolution image parameterization works very well for scintigraphic images of the heart. In coronary artery disease diagnostics we confirm these results as our approach significantly improves on clinical results in terms of diagnostic performance. We improve diagnostic accuracy by 17%, specificity by 12% and sensitivity by 22%. We also significantly improve the number of reliably diagnosed patients by 19% for positive diagnoses, and 16% for negative diagnoses, so that no costly further tests are necessary for them. CONCLUSIONS: Multi-resolution image parameterization equals or even betters that of the physicians in terms of the diagnostic quality of image parameters. By using these parameters for building machine learning classifiers, we can significantly improve diagnostic performance with respect to the results of clinical practice, affect process rationalization, as well as possibly provide novel insights into the diagnostic problems, features and/or processes. PMID- 21646001 TI - Resolution of redundant semantic type assignments for organic chemicals in the UMLS. AB - OBJECTIVE: The Unified Medical Language System (UMLS) integrates terms from different sources into concepts and supplements these with the assignment of one or more high-level semantic types (STs) from its Semantic Network (SN). For a composite organic chemical concept, multiple assignments of organic chemical STs often serve to enumerate the types of the composite's underlying chemical constituents. This practice sometimes leads to the introduction of a forbidden redundant ST assignment, where both an ST and one of its descendants are assigned to the same concept. A methodology for resolving redundant ST assignments for organic chemicals, better capturing the essence of such composite chemicals than the typical omission of the more general ST, is presented. MATERIALS AND METHODS: The typical SN resolution of a redundant ST assignment is to retain only the more specific ST assignment and omit the more general one. However, with organic chemicals, that is not always the correct strategy. A methodology for properly dealing with the redundancy based on the relative sizes of the chemical components is presented. It is more accurate to use the ST of the larger chemical component for capturing the category of the concept, even if that means using the more general ST. RESULTS: A sample of 254 chemical concepts having redundant ST assignments in older UMLS releases was audited to analyze the accuracy of current ST assignments. For 81 (32%) of them, our chemical analysis-based approach yielded a different recommendation from the UMLS (2009AA). New UMLS usage notes capturing rules of this methodology are proffered. CONCLUSIONS: Redundant ST assignments have typically arisen for organic composite chemical concepts. A methodology for dealing with this kind of erroneous configuration, capturing the proper category for a composite chemical, is presented and demonstrated. PMID- 21646002 TI - The Cambridge Breast Intensity-modulated Radiotherapy Trial: patient- and treatment-related factors that influence late toxicity. AB - AIMS: The effect of patient- and treatment-related factors in the development of late normal tissue toxicity after radiotherapy is not yet fully established. The aim of this study was to elucidate the relative importance of such factors in the development of late toxicity after breast-conserving surgery and adjuvant breast radiotherapy. MATERIALS AND METHODS: Patient- and treatment-related factors were analysed in 1014 patients who had received adjuvant radiotherapy to the breast in the Cambridge Breast Intensity-modulated Radiotherapy (IMRT) Trial. Late toxicity data were collected using photographic and clinical assessments and patient reported questionnaires at 2 years after radiotherapy. RESULTS: On multivariate analysis, a larger breast volume was statistically significantly associated with the development of breast shrinkage assessed by serial photographs (odds ratio per litre increase in breast volume = 1.98, 95% confidence interval 1.41, 2.78; P < 0.0005), telangiectasia (odds ratio = 3.94, 95% confidence interval 2.49, 6.24; P < 0.0005), breast oedema (odds ratio = 3.65, 95% confidence interval 2.54, 5.24; P < 0.0005) and pigmentation (odds ratio = 1.75, 95% confidence interval 1.21, 2.51; P = 0.003). Current smokers had an increased risk of developing pigmentation (odds ratio = 2.09, 95% confidence interval 1.23, 3.54; P = 0.006). Patients with a moderate or poor post-surgical cosmesis had a greatly increased risk of moderate or poor overall cosmesis (odds ratio = 38.19; 95% confidence interval 21.9, 66.7; P < 0.0005). Postoperative infection requiring antibiotics was associated with increased risk of telangiectasia (odds ratio = 3.39, 95% confidence interval 1.94, 5.91; P < 0.0005) and breast oversensitivity (odds ratio = 1.78, 95% confidence interval 1.27, 2.49; P = 0.001). CONCLUSIONS: In this study, the greatest risk factors for the development of late toxicity 2 years after breast-conserving surgery and adjuvant radiotherapy were larger breast volume, baseline pre-radiotherapy surgical cosmesis, postoperative infection and possibly smoking. These factors seem to be more important than relatively small differences in dose inhomogeneity and the addition of boost radiotherapy at 2 years after the completion of radiotherapy. The modification of potentially preventable risk factors, such as postoperative infection and smoking, may limit the development of late toxicity after breast radiotherapy. PMID- 21646003 TI - A population-based study of cervix cancer: incidence, management and outcome in the Canadian province of Saskatchewan. AB - AIMS: To document the incidence, management and outcome of uterine cervix cancer in the Canadian province of Saskatchewan. To compare provincial results of low dose rate (LDR) and high dose rate (HDR) brachytherapy in this population. MATERIALS AND METHODS: We carried out a retrospective population-based cohort study of all uterine cervix cancer cases in Saskatchewan diagnosed between 1985 and 2001. We used data from the provincial cancer registry for incident cases, the provincial death registry for vital status information and chart reviews for treatment and toxicity information. RESULTS: In total, 714 cases of uterine cervix cancer were identified. Stage distribution was IA 22.7%, IB 31.8%, II 19.1%, III 18.4%, IV 6.8%; 81.2% were squamous cell carcinoma, 13.4% adenocarcinoma, 1.5% adenosquamous, 0.9% small cell and 3.1% other. The annual crude incidence rate of uterine cervix cancer in Saskatchewan ranged from 6.5 to 12.3% between 1985 and 2001. The 5-year cause-specific survival rate ranged from 100% for stage IA1 to 22% for stage IV. One hundred and seven patients were treated with LDR and 37 with HDR with similar stage distribution. The 5-year cause-specific survival rate was 56% for HDR and 67% for LDR (P = 0.72). For 43 patients managed with external beam radiotherapy alone, outcome was poor for stage IIB to IIIB. The most common acute toxicities of radiation treatment were diarrhoea (60%) and abdominal cramps (12.5%). The common chronic toxicities were vaginal stenosis (5.5%) and small bowel obstruction (4%). CONCLUSIONS: Our population-level outcomes are in keeping with published observed results and provide some of the first Canadian population-level data on HDR and LDR outcomes. We found no significant different in cause-specific survival between patients managed with HDR and LDR, although interpretation is limited by patient numbers. Our results for external beam radiotherapy alone emphasise the vital role brachytherapy plays in the management of cervical cancer. PMID- 21646004 TI - The association between anterior crossbite, deep bite and temporomandibular joint morphology validated by magnetic resonance imaging in an adult non-patient group. AB - The aim of this study was to determine whether associations exist between anterior edge-to-edge bite, anterior crossbite, deep bite and morphology of the temporomandibular joint (TMJ) in the vertical dimension, evaluated by magnetic resonance imaging (MRI) in adults. A total of 148 subjects (90 females, 58 males) were selected from the cross-sectional epidemiological population-based study "Study of Health in Pomerania" (SHIP) for this evaluation. In each person a clinical orthodontical examination was performed. Four linear measurements and one ratio from MRI scans of the TMJs were evaluated. To estimate the effects of anterior edge-to-edge bite, anterior crossbite and deep bite on TMJ variables, linear regression was applied. (Analyses were adjusted for sex, age, school education, and orthodontic treatment.) Anterior edge-to-edge-bite/crossbite had shorter mean eminence heights (eh) on both joint sides (p<0.01). In deep bite without gingival contact the postglenoid process height (pgph) decreased significantly in left joints (p=0.018) and there was a tendency to a decrease in right joints (p=0.059). The ratio between eminence height and postglenoid process height (eh/pgph) was increased on both left (p<0.001) and right (p=0.002) joints. In conclusion, anterior edge-to-edge-bite/crossbite is connected to a reduced eminence height and deep bite without gingival contact with an increased ratio between eminence height and postglenoid process height. Therefore the condylar path in the anterior edge-to-edge bite and crossbite cases can be interpreted to be reduced and in the deep bite cases without gingival contact to be steepened. PMID- 21646005 TI - Enhanced gene replacement frequency in KU70 disruption strain of Stagonospora nodorum. AB - To improve the efficiency of gene disruption in Stagonospora nodorum, the putative KU70 gene encoding the Ku70 protein involved in the nonhomologous end joining double DNA break repair pathway was identified and deleted. The KU70 disruption strain showed no apparent defect in vegetable growth, conidiation and pathogenicity on wheat and barley compared with the wild-type strain. The effect of the absence of KU70 on gene replacement frequency was tested by disruption of TOXA encoding toxin A and LIP2 encoding a putative lipase. Frequency of gene replacement for both genes was dramatically increased in the KU70 disruption strain, compared with the low frequency in the wild-type recipient. PMID- 21646006 TI - Effects of obstructive sleep apnea and its treatment on cardiovascular risk in CAD patients. AB - This study, in optimally treated CAD patients with newly diagnosed OSA, focused on (1) The relationships between OSA and serum biomarkers of four potential pathways of cardiovascular injury in OSA: high-sensitivity C-reactive protein (hs CRP), endothelin-1 (ET-1), N terminal pro B type natriuretic peptide (NT-proBNP) and fibrinogen; and (2) The effect of continuous positive airway pressure (CPAP) therapy on these markers. 151 Chinese patients with proven CAD and standard medication were enrolled. After polysomnography, patients were classified into four groups according to apnea-hypopnea index (AHI): no OSA (n = 25); mild OSA (n = 50); moderate OSA (n = 43); severe OSA (n = 33). Morning levels of hs-CRP, ET 1, NT-proBNP and fibrinogen were assayed and repeated in severe OSA patients after 3-months CPAP treatment. Hs-CRP was greater in patients with severe OSA than those with no OSA or mild OSA (P = 0.001, P = 0.003; respectively). After adjustment for confounders, the hs-CRP levels correlated most strongly with AHI and oxygen desturation index (ODI) (r = 0.439, P < 0.001; r = 0.445, P < 0.001; respectively). In stepwise multiple linear regressions, the strongest predictor of hs-CRP levels was ODI (P < 0.001). After 3 months of CPAP treatment, the hs CRP levels deceased (P = 0.005) in CAD patients with severe OSA. In CAD patients on current optimal medications, hs-CRP is significantly correlated with the severity of OSA, and the elevated hs-CRP levels can be decreased by CPAP. This suggests that OSA could activate vascular inflammation in CAD patients despite current best practice medications. PMID- 21646007 TI - Comparison of airway remodelling assessed by computed tomography in asthma and COPD. AB - BACKGROUND: Few studies have directly compared airway remodelling assessed by computed tomography (CT) between asthma and chronic obstructive pulmonary disease (COPD). The present study was conducted to determine whether there are any differences between the two diseases with similar levels of airflow limitation under clinically stable conditions. METHODS: Subjects included older male asthmatic patients (n = 19) showing FEV(1)/FVC <70% with smoking history less than 5-pack/year. Age- and sex-matched COPD patients (n = 28) who demonstrated similar airflow limitation as asthmatic patients and age-matched healthy non smokers (n = 13) were recruited. Using proprietary software, eight airways were selected in the right lung, and wall area percent (WA%) and airway luminal area (Ai) were measured at the mid-portion of the 3rd to 6th generation of each airway. For comparison, the average of eight measurements per generation was recorded. RESULTS: FEV(1)% predicted and FEV(1)/FVC was similar between asthma and COPD (82.3 +/- 3.3% vs. 77.6 +/- 1.8% and 57.7 +/- 1.6% vs. 57.9 +/- 1.4%). At any generation, WA% was larger and Ai was smaller in asthma, both followed by COPD and then controls. Significant differences were observed between asthma and controls in WA% of the 3rd to 5th generation and Ai of any generation, while no differences were seen between COPD and controls. There were significant differences in Ai of any generation between asthma and COPD. CONCLUSIONS: Airway remodelling assessed by CT is more prominent in asthma compared with age- and sex matched COPD subjects in the 3rd- to 6th generation airways when airflow limitations were similar under stable clinical conditions. PMID- 21646008 TI - A nanoprobe for nonprotein thiols based on assembling of QDs and 4-amino-2,2,6,6 tetramethylpiperidine oxide. AB - A new fluorescent nanoprobe, 4-amino-2,2,6,6-tetramethylpiperidine oxide (AT) functionalized CdTe quantum dots (QDs-AT), was synthesized, for selective detection of nonprotein thiols based on electron transfer (ET). In the presence of nonprotein thiols, the nitroxide radicals in QDs-AT were converted to hydroxylamines, resulting in the fluorescence recovery of the quenched QDs. The detection mechanism of the probe was investigated using Rh-Se-2 probe. The nanoprobe has high sensitivity toward glutathione (GSH) with a detection limit of 7.1 * 10-8 M. The fluorescent imaging of living cells showed that QDs-AT could distinguish the concentration differences of GSH in HL-7702 and HepG2 cells. PMID- 21646009 TI - High-performance electrochemical biosensor for the detection of total cholesterol. AB - We report on a highly sensitive electrochemical biosensor for the determination of total cholesterol. The novel biosensor was fabricated by co-immobilizing three enzymes, cholesterol oxidase (ChO(x)), cholesterol esterase (ChE) and horseradish peroxidase (HRP), on nanoporous gold networks directly grown on a titanium substrate (Ti/NPAu/ChO(x)-HRP-ChE). The morphology and composition of the fabricated nanoporous gold were characterized by scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS) and X-ray diffraction spectroscopy (XRD). The electrochemical behaviour of the Ti/NPAu/ChO(x)-HRP-ChE biosensor was studied using cyclic voltammetry (CV), showing that the developed biosensor possessed high selectivity and high sensitivity (29.33 MUA mM-1 cm-2). The apparent Michaelis-Menten constant, K(M)(app) of this biosensor was very low (0.64 mM), originating from the effective immobilization process and the nanoporous structure of the substrate. The biosensor exhibited a wide linear range up to 300 mg dL-1 in a physiological condition (pH 7.4), which makes it very promising for the clinical determination of cholesterol. The fabricated biosensor was further tested using real food samples margarine, butter and fish oil, showing that the biosensor has the potential to be used as a facile cholesterol detection tool in food and supplement quality control. PMID- 21646010 TI - Microbial production of diols as platform chemicals: recent progresses. AB - Diols are chemicals with two hydroxyl groups which have a wide range of appealing applications as chemicals and fuels. In particular, four diol compounds, namely 1,3-propanediol (1,3-PDO), 1,2-propanediol (1,2-PDO), 2,3-butanediol (2,3-BDO) and 1,4-butanediol (1,4-BDO) can be biotechnologically produced by direct microbial bioconversion of renewable materials. These diols are considered as platform green chemicals. We review and discuss here the recent development in the microbial production of these diols, especially regarding the engineering of production strains and optimization of the fermentation processes. PMID- 21646011 TI - In vivo tissue engineering of musculoskeletal tissues. AB - Tissue engineering of musculoskeletal tissues often involves the in vitro manipulation and culture of progenitor cells, growth factors and biomaterial scaffolds. Though in vitro tissue engineering has greatly increased our understanding of cellular behavior and cell-material interactions, this methodology is often unable to recreate tissue with the hierarchical organization and vascularization found within native tissues. Accordingly, investigators have focused on alternative in vivo tissue engineering strategies, whereby the traditional triad (cells, growth factors, scaffolds) or a combination thereof are directly implanted at the damaged tissue site or within ectopic sites capable of supporting neo-tissue formation. In vivo tissue engineering may offer a preferential route for regeneration of musculoskeletal and other tissues with distinct advantages over in vitro methods based on the specific location of endogenous cultivation, recruitment of autologous cells, and patient-specific regenerated tissues. PMID- 21646012 TI - Sound localization: Jeffress and beyond. AB - Many animals use the interaural time differences (ITDs) to locate the source of low frequency sounds. The place coding theory proposed by Jeffress has long been a dominant model to account for the neural mechanisms of ITD detection. Recent research, however, suggests a wider range of strategies for ITD coding in the binaural auditory brainstem. We discuss how ITD is coded in avian, mammalian, and reptilian nervous systems, and review underlying synaptic and cellular properties that enable precise temporal computation. The latest advances in recording and analysis techniques provide powerful tools for both overcoming and utilizing the large field potentials in these nuclei. PMID- 21646013 TI - Neuromodulation and flexibility in Central Pattern Generator networks. AB - Central Pattern Generator (CPG) networks, which organize rhythmic movements, have long served as models for neural network organization. Modulatory inputs are essential components of CPG function: neuromodulators set the parameters of CPG neurons and synapses to render the networks functional. Each modulator acts on the network by many effects which may oppose one another; this may serve to stabilize the modulated state. Neuromodulators also determine the active neuronal composition in the CPG, which varies with state changes such as locomotor speed. The pattern of gene expression which determines the electrophysiological personality of each CPG neuron is also under modulatory control. It is not possible to model the function of neural networks without including the actions of neuromodulators. PMID- 21646014 TI - Visual perception and saccadic eye movements. AB - We use saccades several times per second to move the fovea between points of interest and build an understanding of our visual environment. Recent behavioral experiments show evidence for the integration of pre- and postsaccadic information (even subliminally), the modulation of visual sensitivity, and the rapid reallocation of attention. The recent physiological literature has identified a characteristic modulation of neural responsiveness-perisaccadic reduction followed by a postsaccadic increase-that is found in many visual areas, but whose source is as yet unknown. This modulation seems optimal for reducing sensitivity during and boosting sensitivity between saccades, but no study has yet established a direct causal link between neural and behavioral changes. PMID- 21646015 TI - Adenoviral vector expressing short hairpin RNA targeting Wnt2B has an effective antitumour activity against Wnt2B2-overexpressing tumours. AB - BACKGROUND: The Wnt family encodes multi-functional signalling glycoproteins regulating various normal and pathological processes including tumourigenesis. Wnt2B overexpression is thought to affect tumour progression through the activation of the canonical Wnt pathway. METHOD: Experimental studies were conducted using a Wnt2B-inhibiting vector to establish gene therapy against Wnt2B2-overexpressing tumours. A replication-deficient recombinant adenoviral vector expressing short hairpin RNA targeting Wnt2B (Ad-shWnt2B) was constructed. Three Wnt2B2-overexpressing human tumour cells, including A549 cells, Hela cells and PANC1 cells, were used. Thereafter, cell viability was evaluated using 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Next, a human tumour xenograft model in nude mice was prepared by subcutaneously implanting tumours derived from A549 cells. Ad-shWnt2B was administered via intratumoural injection every 4days. RESULTS: First, immunohistochemical studies revealed that high levels of Wnt2B expression appeared in proliferative normal tissues and many human tumour tissues. Furthermore, the Wnt2B2 gene expression was associated with c-Myc and survivin expressions in human lung cancer. Transduction with Ad-shWnt2B effectively downregulated the Wnt2B2 expression in all the three Wnt2B2 overexpressing tumour cells (p<0.0001). The transduction with Ad-shWnt2B significantly reduced the percentage of viable cells in all the Wnt2B2 overexpressing tumour cells (p<0.005). In addition, transduction with Ad-shWnt2B significantly downregulated c-Myc and survivin in A549 cells (p<0.005). Furthermore, the treatment with Ad-shWnt2B exerted a significant antitumour effect against the Wnt2B2-overexpressing A549 xenografts by inducing apoptosis (p<0.01). CONCLUSIONS: Cancer gene therapy using an adenoviral vector expressing short hairpin RNA (shRNA) against Wnt2B was, therefore, found to have a strong antitumour effect against Wnt2B2-overexpressing tumours. PMID- 21646016 TI - Development of low-concentration mercury adsorbents from biohydrogen-generation agricultural residues using sulfur impregnation. AB - Mercury adsorbents were derived from waste biohydrogen-generation barley husk and rice husk via carbonization, steam activation, and sulfur impregnation at 300-650 degrees C. The samples derived from agricultural residues showed a greater Hg(0) adsorption than that of a coal-based activated carbon, confirming the feasibility of resource recovery of these agricultural residuals for low-concentration gaseous Hg adsorption. Sulfur impregnation reduced both the surface area and pore volume of the samples, with lower temperature causing a greater decrease. Elevating the impregnation temperature increased the organic sulfur contents, suggesting that in addition to elemental sulfur, organic sulfur may also act as active sites to improve Hg(0) adsorption. Oxygen and sulfur functional groups accompanying the microporous structures may account for the enhancing Hg(0) adsorption of the raw and sulfur-treated samples, respectively. The pseudo-second order model can best describe the chemisorption characteristics, implying that Hg(0) adsorption on the samples was in a bimolecular reaction form. PMID- 21646017 TI - Heat of reaction measurements for hydrothermal carbonization of biomass. AB - This paper presents a set of calorimetric measurements with the aim of better understanding the calorific nature of hydrothermal carbonization. Presented values so far show an inadequately high scatter to do so, preventing a well funded assessment of the energetic feasibility of this process. The heat released during hydrothermal carbonization at 240 degrees C measured with the applied differential calorimetry setup is -1.06MJ/kg(glucose,daf) with a standard deviation of 14%, -1.07MJ/kg(cellulose,daf) with a standard deviation of 9%, and 0.76MJ/kg(wood,daf) with a standard deviation of 32%. These results are in good agreement with the theoretically derived maximum heat release. Despite the comparably high experimental standard deviation of these results, their accuracy is considerably higher than previously published results. PMID- 21646018 TI - Fungal pretreatment: An alternative in second-generation ethanol from wheat straw. AB - The potential of a fungal pretreatment combined with a mild alkali treatment to replace or complement current physico-chemical methods for ethanol production from wheat straw has been investigated. Changes in substrate composition, secretion of ligninolytic enzymes, enzymatic hydrolysis efficiency and ethanol yield after 7, 14 and 21 days of solid-state fermentation were evaluated. Most fungi degraded lignin with variable selectivity degrees, although only eight of them improved sugar recovery compared to untreated samples. Glucose yield after 21 days of pretreatment with Poria subvermispora and Irpex lacteus reached 69% and 66% of cellulose available in the wheat straw, respectively, with an ethanol yield of 62% in both cases. Conversions from glucose to ethanol reached around 90%, showing that no inhibitors were generated during this pretreatment. No close correlations were found between ligninolytic enzymes production and sugar yields. PMID- 21646019 TI - Studying thermal characteristics of seating materials by recording temperature from 3 positions at the seat-subject interface. AB - AIM OF THE STUDY: To determine whether 3 fixed positions of seat-subject interface temperature measurement offer more information than a single point of measurement. MATERIALS AND METHODS: Temperature data was simultaneously acquired (sampling frequency 1 Hz/sensor) from each of three sensor positions (right & left mid-thigh and coccyx), from the subject-seat interface. The data was acquired whilst subjects (6 males, 5 females: 21-40 yrs: BMI 19.3-26.4) sat for 20 min on each of three types of seat material (foam, gel mould and solid wood). Data collection was performed at the same time of day for each subject: ambient temperature between 21.1 and 21.2 degrees C, ambient relative humidity 50.9%. RESULTS: Analysis of data from the sensors, post mathematical smoothing, for each subject (n = 11; ANOVA, followed by post-hoc t-tests) revealed each of the measurement positions to have a significantly different recorded temperature (p < 0.01). However, profile of temperature change at the same measurement position using the same seating material during the 20 min sitting period, was highly correlated (r > 0.99) between subjects, a consistent finding across all 11 subjects regardless of seat material selected. CONCLUSION: Use of 3 positions of measurement (3 sensors) appears necessary when performing detailed studies of temperature change at the seat-subject interface. The high level of comparability of results between subjects supports potential of this method to resolve quantitative components of qualitative measurements, e.g., thermal comfort. PMID- 21646020 TI - Impairment in postural control is greater when ankle plantarflexors and dorsiflexors are fatigued simultaneously than when fatigued separately. AB - The aim of the present study was to compare the effects of ankle muscle fatigue on postural control when plantarflexors (PFs) and dorsiflexors (DFs) are fatigued simultaneously compared with separately. This study also investigated the recovery of postural control after fatigue. Sixteen adults (eight women and eight men) performed postural trials before and after an isokinetic fatigue task involving either (i) only PFs (30 degrees s(-1)), (ii) only DFs (120 degrees s( 1)), or (iii) both PFs and DFs simultaneously. The fatigue task involved maximal contractions repeated until the torque produced decreased below 50% of the maximal torque. Postural trials lasted 30s and were performed on one leg with eyes open (EO) or eyes closed (EC). Sway area, medio-lateral (ML) and antero posterior (AP) positions and velocities were calculated from the center of pressure displacements. With EO, no effect of fatigue was found on postural variables. With EC, sway area and AP velocity increased only when both PFs and DFs were fatigued simultaneously. An effect of fatigue present only when both muscle groups are fatigued simultaneously could be due to impairment in the compensatory activity between agonist and antagonist muscles and/or a greater decrease in proprioception due to a greater number of fatigued muscles. In addition, when PFs and DFs were fatigued simultaneously, sway area and AP velocity returned to pre-fatigue values within 2min, whereas a posterior shift in AP position persisted for 10min. This last result may suggest a longer-lasting change in postural strategy needed for optimal postural control. PMID- 21646021 TI - Postural control after traumatic brain injury in patients with neuro-ophthalmic deficits. AB - Postural instability is a common and devastating consequence of traumatic brain injury (TBI). The majority of TBI patients also suffer from neuro-ophthalmic deficits that can be an important contributing element to their sensation of vertigo and dizziness. Static posturography aims at the objective evaluation of patient balance impairment, but is usually affected by large inter- and intra subject variability. Here we propose a protocol based on 10 randomized trials stimulating in different ways the visual and vestibular systems. Due to its completeness, our protocol highlights the specific residual difficulties of each patient in the various conditions. In this way, it was possible to evidence significant balance abnormalities in TBI patients with respect to controls. Moreover, by means of a multivariate analysis we were able to discriminate different levels of residual neuro-ophthalmic impairment. PMID- 21646022 TI - Efficacy of clinical gait analysis: A systematic review. AB - The aim of this systematic review was to evaluate and summarize the current evidence base related to the clinical efficacy of gait analysis. A literature review was conducted to identify references related to human gait analysis published between January 2000 and September 2009 plus relevant older references. The references were assessed independently by four reviewers using a hierarchical model of efficacy adapted for gait analysis, and final scores were agreed upon by at least three of the four reviewers. 1528 references were identified relating to human instrumented gait analysis. Of these, 116 original articles addressed technical accuracy efficacy, 89 addressed diagnostic accuracy efficacy, 11 addressed diagnostic thinking and treatment efficacy, seven addressed patient outcomes efficacy, and one addressed societal efficacy, with some of the articles addressing multiple levels of efficacy. This body of literature provides strong evidence for the technical, diagnostic accuracy, diagnostic thinking and treatment efficacy of gait analysis. The existing evidence also indicates efficacy at the higher levels of patient outcomes and societal cost effectiveness, but this evidence is more sparse and does not include any randomized controlled trials. Thus, the current evidence supports the clinical efficacy of gait analysis, particularly at the lower levels of efficacy, but additional research is needed to strengthen the evidence base at the higher levels of efficacy. PMID- 21646023 TI - Force probing cell shape changes to molecular resolution. AB - Atomic force microscopy (AFM) is a force sensing nanoscopic tool that can be used to undertake a multiscale approach to understand the mechanisms that underlie cell shape change, ranging from the cellular to molecular scale. In this review paper, we discuss the use of AFM to characterize the dramatic shape changes of mitotic cells. AFM-based mechanical assays can be applied to measure the considerable rounding force and hydrostatic pressure generated by mitotic cells. A complementary AFM technique, single-molecule force spectroscopy, is able to quantify the interactions and mechanisms that functionally regulate individual proteins. Future developments of these nanomechanical methods, together with advances in light microscopy imaging and cell biological and genetic tools, should provide further insight into the biochemical, cellular and mechanical processes that govern mitosis and other cell shape change phenomena. PMID- 21646024 TI - Survivorship and patient satisfaction of a fixed bearing unicompartmental knee arthroplasty incorporating an all-polyethylene tibial component. AB - We report the survivorship of 91 fixed bearing unicompartmental arthroplasties with all-polyethylene tibial components (Preservation DePuy UK), which were used for medial compartment osteoarthritis in 79 patients between 2004 and 2007. The satisfaction level of patients who had not undergone revision of the implant was also recorded. For comparison, we reviewed 49 mobile bearing unicompartmental arthroplasties (Oxford UKA Biomet UK Ltd), which had been used in 44 patients between 1998 and 2007. Mean length of follow-up of patients with the fixed bearing implant was 44.7 months (range 24-74 months) and for the mobile bearing replacement, the mean follow-up was 67.6 months (24-119). In the fixed bearing design, at maximum follow-up period of 74 months, eight implants (8.8%) had been revised (or were listed for revision) to Total Knee Replacement and in the mobile bearing design over the maximum follow-up period of 119 months there had been only one revision (2.0%). Patients who had not undergone revision were asked if they were satisfied with their knee following the unicompartmental arthroplasty. In the fixed bearing design, 83.5% said that they were satisfied with the outcome of the operation compared to 93.9% of the patients receiving the mobile bearing design. We conclude that there is a higher incidence of revision of this fixed bearing design using an all-polyethylene tibial component compared to the mobile bearing design. We found that those patients who had not required revision had a lower rate of satisfaction with the fixed bearing compared to the mobile bearing design. PMID- 21646025 TI - The use of standard operating procedures in day case anterior cruciate ligament reconstruction. AB - The current rate of day-case anterior cruciate ligament reconstruction (ACLR) in the UK remains low. Although specialised care pathways with standard operating procedures (SOPs) have been effective in reducing length of stay following some surgical procedures, this has not been previously reported for ACLR. We evaluate the effectiveness of SOPs for establishing day-case ACLR in a specialist unit. Fifty patients undergoing ACLR between May and September 2010 were studied prospectively ("study group"). SOPs were designed for pre-operative assessment, anaesthesia, surgical procedure, mobilisation and discharge. We evaluated length of stay, readmission rates, patient satisfaction and compliance to SOPs. A retrospective analysis of 50 patients who underwent ACLR prior to implementation of the day-case pathway was performed ("standard practice group"). Eighty percent of patients in the study group were discharged on the day of surgery (mean length of stay=5.3h) compared to 16% in the standard practice group (mean length of stay=21.6h). This difference was statistically significant (p<0.05, Mann-Whitney U test). All patients were satisfied with the day case pathway. Ninety-two percent of the study group were discharged on the day of surgery when all SOPs were followed and 46% where they were not. High rates of day-case ACLR with excellent patient satisfaction can be achieved with the use of a specialised patient pathway with SOPs. PMID- 21646026 TI - Neutron flux measurements at the TRIGA reactor in Vienna for the prediction of the activation of the biological shield. AB - The activation of the biological shield is an important process for waste management considerations of nuclear facilities. The final activity can be estimated by modeling using the neutron flux density rather than the radiometric approach of activity measurements. Measurement series at the TRIGA reactor Vienna reveal that the flux density next to the biological shield is in the order of 10(9)cm(-2)s(-1) at maximum power; but it is strongly influenced by reactor installations. The data allow the estimation of the final waste categorization of the concrete according to the Austrian legislation. PMID- 21646027 TI - Production and separation of no-carrier-added thallium isotopes from proton irradiated (nat)Hg2Cl2 matrix. AB - For the first time, (nat)Hg2Cl2 target has been used to produce no-carrier-added NCA (197,198,198m,199,200,201)Tl radionuclides using (nat)Hg(p,xn) reaction. Liquid-liquid extraction technique was employed in order to separate radiothallium from the bulk mercury matrix using liquid anion exchanger trioctylamine (TOA) dissolved in cyclohexane. In order to verify the presence of stable Hg in Tl fraction, the entire process was repeated with stable salts of Hg and Tl and the extent of separation was examined by Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES). High separation factors were observed both by radiometric and ICP-OES technique when 0.1 M HNO3 and 0.1M TOA were used as aqueous and organic phase, respectively. The Hg contamination was less than 0.3 ppm in the aqueous phase containing Tl after three times of extraction at the optimal condition. PMID- 21646028 TI - Metabolic nuclear receptor signaling and the inflammatory acute phase response. AB - The acute phase response (APR) classically refers to the rapid reprogramming of gene expression and metabolism in response to inflammatory cytokine signaling. As components of the innate immune system, hepatocyte-derived acute phase proteins (APPs) play a central role in restoring tissue homeostasis. Recently, an intriguing 'metaflammatory' facet of the APR became evident with chronically elevated APP levels being connected to metabolic syndrome disorders. The causality of these connections is unclear but could relate to adverse metabolic and inflammatory disturbances, particularly those affecting lipoprotein properties, cholesterol metabolism and atherogenesis. Here we review these aspects with an emphasis on the emerging importance of lipid-sensing nuclear receptors (LXRs, LRH-1, PPARs), in conjunction with anti-inflammatory transrepression pathways, as physiological and pharmacological relevant modulators of the APR. PMID- 21646029 TI - Endovascular stenting of extracranial carotid artery aneurysm: a systematic review. AB - OBJECTIVE: The objective of this study was to systematically review all available published data about endovascular stenting for patients with extracranial carotid artery aneurysm (ECAA). METHODS: All clinical reports in the English language on endovascular stenting for ECAA from 1995 through 2010 were identified. General clinical information, aetiology and parameter of carotid artery aneurysm, information regarding devices, indications for endovascular stenting and in hospital and follow-up data of each patient were extracted individually. RESULTS: A total of 113 studies, involving a total of 224 patients, were included. Procedure success was reported in 92.8% of patients. Postoperative endoleak was reported in 8.1% of patients. The incidence of stroke was 1.8%. Cranial nerve injury occurred in 0.5%. Overall in-hospital mortality was 4.1%. The mean follow up was 15.4 +/- 15.3 months. Stent-graft patency rate was 93.2%. CONCLUSION: Endovascular stenting is technically feasible with high procedure success and relatively low complication rate in patients with ECAA. Both short- and mid-term outcomes appear to be favourable. PMID- 21646030 TI - Self reported pain severity among multiethnic older Singaporeans: does adjusting for reporting heterogeneity matter? AB - The objective of this paper is to test and correct for systematic differences in reporting of pain severity among older adults by age, gender, ethnic group and socio-economic status using anchoring vignettes. Data from a national survey of community-dwelling older Singaporeans (aged 60 years and over) conducted in 2009 was used. Respondents were asked to rate the severity of their own pain as well as that of others described in the vignettes on a five-point scale ranging from none to extreme. An ordered probit model was used to estimate the coefficients of the independent variables (age, gender, ethnic group, education, housing type) on self-reported pain. Reporting heterogeneity in pain severity was then corrected using a Hierarchical Ordered Probit model. The results showed that before correcting for reporting heterogeneity, women, those older, and those of Malay ethnicity reported greater severity of pain, while there was no association of reported pain severity with housing type and education. However, after correcting for reporting heterogeneity, while women and those older were found to have an even greater severity of pain than what they had reported, Malays were found to have a lower severity of pain than what they had reported. We conclude that there are systematic differences in reporting pain severity by age, gender and ethnic group. We propose that pain management may be improved if medical professionals take into account reporting heterogeneity for pain severity among various population sub-groups in Singapore. PMID- 21646031 TI - Liver glycogen storage diseases due to phosphorylase system deficiencies: diagnosis thanks to non invasive blood enzymatic and molecular studies. AB - Glycogen storage disease (GSD) due to a deficient hepatic phosphorylase system defines a genetically heterogeneous group of disorders that mainly manifests in children. We investigated 45 unrelated children in whom a liver GSD VI or IX was suspected on the basis of clinical symptoms including hepatomegaly, increased serum transaminases, postprandial lactatemia and/or mild fasting hypoglycemia. Liver phosphorylase and phosphorylase b kinase activities studied in peripheral blood cells allowed to suspect diagnosis in 37 cases but was uninformative in 5. Sequencing of liver phosphorylase genes was useful to establish an accurate diagnosis. Causative mutations were found either in the PYGL (11 patients), PHKA2 (26 patients), PHKG2 (three patients) or in the PHKB (three patients) genes. Eleven novel disease causative mutations, five missense (p.N188K, p.D228Y, p.P382L, p.R491H, p.L500R) and six truncating mutations (c.501_502ins361pb, c.528+2T>C, c.856-29_c.1518+614del, c.1620+1G>C, p.E703del and c.2313-1G>T) were identified in the PYGL gene. Seventeen novel disease causative mutations, ten missense (p.A42P, p.Q95R, p.G131D, p.G131V, p.Q134R, p.G187R, p.G300V, p.G300A, p.C326Y, p.W820G) and seven truncating (c.537+5G>A, p.G396DfsX28, p.Q404X, p.N653X, p.L855PfsX87, and two large deletions) were identified in the PHKA2 gene. Four novel truncating mutations (p.R168X, p.Q287X, p.I268PfsX12 and c.272 1G>C) were identified in the PHKG2 gene and three (c.573_577del, p.R364X, c.2427+3A>G) in the PHKB gene. Patients with PHKG2 mutations evolved towards cirrhosis. Molecular analysis of GSD VI or IX genes allows to confirm diagnosis suspected on the basis of enzymatic analysis and to establish diagnosis and avoid liver biopsy when enzymatic studies are not informative in blood cells. PMID- 21646032 TI - Safety of extended treatment with sapropterin dihydrochloride in patients with phenylketonuria: results of a phase 3b study. AB - BACKGROUND: Phenylketonuria (PKU) results from impaired breakdown of phenylalanine (Phe) due to deficient phenylalanine hydroxylase (PAH) activity. Sapropterin dihydrochloride (sapropterin, Kuvan(r)) is the only US- and EU approved pharmaceutical version of naturally occurring 6R-BH(4), the cofactor required for PAH activity. Sapropterin enhances residual PAH activity in sapropterin-responsive PKU patients and, in conjunction with dietary management, helps reduce blood Phe concentrations for optimal control. Approval was based on the positive safety and efficacy results of four international clinical studies, the longest of which was 22 weeks in duration. OBJECTIVE: To evaluate the safety of long-term treatment with sapropterin in PKU subjects who participated in previous Phase 3 sapropterin trials. METHODS: PKU-008 was designed as a Phase 3b, multicenter, multinational, open-label, 3-year extension trial to evaluate the long-term safety of sapropterin in patients with PKU who were classified as sapropterin responders and participated in prior Phase 3 sapropterin studies: 111 subjects aged 4-50 years completed prior studies and were subsequently enrolled in study PKU-008. Routine safety monitoring was performed at 3-month intervals and included adverse event reporting, blood Phe monitoring, clinical laboratory evaluations, physical examinations and vital sign measurements. RESULTS: Average exposure during PKU-008 was 658.7+/-221.3 days (range, 56-953; median, 595). The average total duration of participation in multiple studies (PKU-001, PKU-003, PKU-004, and PKU-008; or PKU-006 and PKU-008) was 799.0+/-237.5 days (range, 135 1151). The mean sapropterin dose was 16.2+/-4.7 mg/kg/day. Most adverse events were considered unrelated to treatment, were mild or moderate in severity, and were consistent with prior studies of sapropterin. No age-specific differences were observed in adverse event reporting. Three subjects discontinued treatment due to adverse events that were considered possibly or probably related to study treatment (one each of difficulty concentrating, decreased platelet count, and intermittent diarrhea). No deaths were reported. Of seven reported serious adverse events, one was considered possibly related to study treatment (gastroesophageal reflux). There were no laboratory or physical examination abnormalities requiring medical interventions. For most subjects, blood Phe concentrations were consistently within target range, confirming the durability of response in subjects undergoing extended treatment with sapropterin. CONCLUSION: Sapropterin treatment was found to be safe and well tolerated at doses of 5 to 20mg/kg/day for an average exposure of 659 days. This study supports the safety and tolerability of sapropterin as long-term treatment for patients with PKU. PMID- 21646034 TI - Iron removal from kaolin using thiourea assisted by ultrasonic wave. AB - In the present study a bleaching process of a kaolinite was carried out using thiourea as the leachant agent in the iron removal process, in the absence and presence of ultrasound. The effect of thiourea was investigated together with other factors, such as thiourea concentration, temperature, treatment time, and ultrasonic parameters. The optimum conditions for the maximum whiteness of 89% with ultrasound were determined as follows: reaction temperature, 20 degrees C; ultrasound frequency, 80 kHz; ultrasound power, 500 W; thiourea concentration, 0.4 wt.%; pH, 3.0; reaction time, 20 min. The assistance of ultrasound led to a remarkable acceleration for the iron leaching process, and dramatic reduction in the concentration of leach reagent, irradiation time, and reaction temperature, when compared with the conventional bleaching method using thiourea in the absence of ultrasound. PMID- 21646035 TI - Names-based classification of accident and emergency department users. AB - This paper studies differential healthcare utilisation by ethnic group. Administrative records of nearly 100,000 users of an Inner London accident and emergency (A&E) facility were analysed using an innovative names-based ethnicity classification. Adult repeated 'light' usage (i.e. with no hospitalisation or follow-up) did not differ according to ethnic group. Users from ethnic minorities had lower GP registration rates than the majority reference group. However, lack of GP registration was not associated with repeated light use of A&E, overall. Therefore, these results challenge common perceptions of differential A&E access rates by ethnicity. PMID- 21646036 TI - The double crush syndrome revisited--a Delphi study to reveal current expert views on mechanisms underlying dual nerve disorders. AB - A high prevalence of dual nerve disorders is frequently reported. How a secondary nerve disorder may develop following a primary nerve disorder remains largely unknown. Although still frequently cited, most explanatory theories were formulated many years ago. Considering recent advances in neuroscience, it is uncertain whether these theories still reflect current expert opinion. A Delphi study was conducted to update views on potential mechanisms underlying dual nerve disorders. In three rounds, seventeen international experts in the field of peripheral nerve disorders were asked to list possible mechanisms and rate their plausibility. Mechanisms with a median plausibility rating of >=7 out of 10 were considered highly plausible. The experts identified fourteen mechanisms associated with a first nerve disorder that may predispose to the development of another nerve disorder. Of these fourteen mechanisms, nine have not previously been linked to double crush. Four mechanisms were considered highly plausible (impaired axonal transport, ion channel up or downregulation, inflammation in the dorsal root ganglia and neuroma-in-continuity). Eight additional mechanisms were listed which are not triggered by a primary nerve disorder, but may render the nervous system more vulnerable to multiple nerve disorders, such as systemic diseases and neurotoxic medication. Even though many mechanisms were classified as plausible or highly plausible, overall plausibility ratings varied widely. Experts indicated that a wide range of mechanisms has to be considered to better understand dual nerve disorders. Previously listed theories cannot be discarded, but may be insufficient to explain the high prevalence of dual nerve disorders. PMID- 21646037 TI - Response to Keeley et al.: Fire as an evolutionary pressure shaping plant traits. PMID- 21646038 TI - An integrated approach to segmentation and nonrigid registration for application in image-guided pelvic radiotherapy. AB - External beam radiotherapy (EBRT) has become the preferred options for nonsurgical treatment of prostate cancer and cervix cancer. In order to deliver higher doses to cancerous regions within these pelvic structures (i.e. prostate or cervix) while maintaining or lowering the doses to surrounding non-cancerous regions, it is critical to account for setup variation, organ motion, anatomical changes due to treatment and intra-fraction motion. In previous work, manual segmentation of the soft tissues is performed and then images are registered based on the manual segmentation. In this paper, we present an integrated automatic approach to multiple organ segmentation and nonrigid constrained registration, which can achieve these two aims simultaneously. The segmentation and registration steps are both formulated using a Bayesian framework, and they constrain each other using an iterative conditional model strategy. We also propose a new strategy to assess cumulative actual dose for this novel integrated algorithm, in order to both determine whether the intended treatment is being delivered and, potentially, whether or not a plan should be adjusted for future treatment fractions. Quantitative results show that the automatic segmentation produced results that have an accuracy comparable to manual segmentation, while the registration part significantly outperforms both rigid and nonrigid registration. Clinical application and evaluation of dose delivery show the superiority of proposed method to the procedure currently used in clinical practice, i.e. manual segmentation followed by rigid registration. PMID- 21646039 TI - Active deformation fields: dense deformation field estimation for atlas-based segmentation using the active contour framework. AB - This paper presents a new and original variational framework for atlas-based segmentation. The proposed framework integrates both the active contour framework, and the dense deformation fields of optical flow framework. This framework is quite general and encompasses many of the state-of-the-art atlas based segmentation methods. It also allows to perform the registration of atlas and target images based on only selected structures of interest. The versatility and potentiality of the proposed framework are demonstrated by presenting three diverse applications: In the first application, we show how the proposed framework can be used to simulate the growth of inconsistent structures like a tumor in an atlas. In the second application, we estimate the position of nonvisible brain structures based on the surrounding structures and validate the results by comparing with other methods. In the final application, we present the segmentation of lymph nodes in the Head and Neck CT images, and demonstrate how multiple registration forces can be used in this framework in an hierarchical manner. PMID- 21646040 TI - Mouth self-examination to improve oral cancer awareness and early detection in a high-risk population. AB - Oral cancer is a potentially preventable disease due to its association with well known risk factors and easy detectability. There is a significant deficiency in the awareness of oral cancer and its risk factors among the public. Raising public awareness could effectively contribute to achieving a significant reduction in the incidence of oral cancer. The objective of this study was to evaluate the effectiveness of mouth self-examination (MSE) in improving the awareness of oral cancer and its risk factors as well as test its feasibility as an oral cancer-screening tool. The study was carried out in a high-risk population of 57,704 from India, of which, 34,766 individuals who have met the eligibility criteria formed the study population. MSE brochures and trained health workers were employed for the purpose of health education and cancer screening. The present study compared their efficacy to detect oral lesions. Subjects with suspicious lesions were referred to the trained oral cancer specialist for confirmation. A questionnaire to assess the awareness of oral cancer and its risk factors was developed and validated. SPSS (v.11.0) was used for data analysis. The program identified 216 cases of potentially malignant lesions as well as three cases of oral cancer. The findings of MSE and health workers showed 72% concordance, while that of health workers and oral cancer specialist showed 100% concordance. MSE had a low sensitivity of 18%, while the specificity was 99.9%. Though the technique identified high-risk lesions such as red patches (66.7%) and non-healing ulcers (42.9%), the detection rate of white patches was low (12.7%). Overall awareness of oral cancer and its risk factors after introduction of MSE program was over 80%; but the compliance to seek treatment was poor (32%). Mouth self-examination may be used as an effective tool to improve the awareness of oral cancer and for the early detection of lesions. PMID- 21646041 TI - Vibrational spectroscopic studies and DFT calculations on tribromoacetate and tribromoacetic acid in aqueous solution. AB - Aqueous solutions of sodium tribromoacetate (NaCBr3CO2) and its corresponding acid (CBr3COOH) have been studied using Raman and infrared spectroscopy. The spectra of the species in solution were assigned according to symmetry Cs. Characteristic bands of CBr3CO2-(aq) and the tribromoacetic acid, CBr3COOH(aq), are discussed. For the hydrated anion, the CO2 group, the symmetric CO2 stretching mode at 1332 cm(-1) and the asymmetric stretching mode at 1651 cm(-1) are characteristic while the CO mode at 1730 cm(-1) is characteristic for the spectra of the acid. The stretching mode, nuC-C at 912cm(-1) for CBr3CO2-(aq) is 10 cm(-1) lower in the anion compared with that of the acid. These characteristic modes are compared to those in acetate, CH3CO2-(aq). Coupling of the modes are fairly extensive and therefore DFT calculations have been carried out in order to compare the measured spectra with the calculated ones. The geometrical parameters such as bond length and bond angles of the tribromoacetate, and tribromoacetic acid have been obtained and may be compared with the ones published for other acetates and their conjugated acids. CBr3COOH(aq) is a moderately strong acid and the pKa value derived from quantitative Raman measurements is equal to -0.23 at 23 degrees C. The deuterated acid CBr3COOD in heavy water has been measured as well and the assignments were given. PMID- 21646042 TI - A vibrational spectroscopic study of the mixed anion mineral sanjuanite Al2(PO4)(SO4)(OH).9H2O. AB - The mineral sanjuanite Al2(PO4)(SO4)(OH).9H2O has been characterised by Raman spectroscopy complimented by infrared spectroscopy. The mineral is characterised by an intense Raman band at 984 cm(-1), assigned to the (PO4)3- nu1 symmetric stretching mode. A shoulder band at 1037 cm(-1) is attributed to the (SO4)2- nu1 symmetric stretching mode. Two Raman bands observed at 1102 and 1148 cm(-1) are assigned to (PO4)3- and (SO4)2- nu3 antisymmetric stretching modes. Multiple bands provide evidence for the reduction in symmetry of both anions. This concept is supported by the multiple sulphate and phosphate bending modes. Raman spectroscopy shows that there are more than one non-equivalent water molecules in the sanjuanite structure. There is evidence that structural disorder exists, shown by the complex set of overlapping bands in the Raman and infrared spectra. At least two types of water are identified with different hydrogen bond strengths. The involvement of water in the sanjuanite structure is essential for the mineral stability. PMID- 21646043 TI - Study on the toxic interaction of methanol, ethanol and propanol against the bovine hemoglobin (BHb) on molecular level. AB - The toxic interaction of methanol, ethanol and propanol with bovine hemoglobin (BHb) at protein molecular level was studied by resonance light scattering (RLS), fluorescence, ultraviolet-visible absorption (UV-vis) and circular dichroism (CD) techniques. The experimental results showed that the three alcohols all had toxic effects on BHb and the effects increased along with the increasing alcohol dose. The results of RLS and fluorescence spectroscopy showed that alcohols can denature BHb. They changed the microenvironment of amino acid residues and led to molecular aggregation. The decreasing order of the influence is propanol, ethanol and methanol. The results of UV-vis and CD spectra revealed that alcohols led to conformational changes of BHb, including the loosening of the skeleton structure and the decreasing of alpha-helix in the second structure. The changes generated by propanol were much larger than those by methanol and ethanol. PMID- 21646044 TI - Structural and spectroscopic characterization of 2,3-difluorobenzoic acid and 2,4 difluorobenzoic acid with experimental techniques and quantum chemical calculations. AB - In this study, the molecular conformation, vibrational and electronic transition analysis of 2,3-difluorobenzoic acid and 2,4-difluorobenzoic acid (C7H4F2O2) were presented using experimental techniques (FT-IR, FT-Raman and UV) and quantum chemical calculations. FT-IR and FT-Raman spectra in solid state were recorded in the region 4000-400 cm(-1) and 4000-5 cm(-1), respectively. The UV absorption spectra of the compounds that dissolved in ethanol were recorded in the range of 200-800 nm. The structural properties of the molecules in the ground state were calculated using density functional theory (DFT) and second order Moller-Plesset perturbation theory (MP2) employing 6-311++G(d,p) basis set. Optimized structure of compounds was interpreted and compared with the earlier reported experimental values. The scaled vibrational wavenumbers were compared with experimental results. The complete assignments were performed on the basis of the experimental data and total energy distribution (TED) of the vibrational modes, calculated with scaled quantum mechanics (SQM) method. A study on the electronic properties, such as absorption wavelength, excitation energy, dipole moment and frontier molecular orbital energy, were performed by time dependent DFT (TD-DFT) approach. Based on the UV spectra and TD-DFT calculations, the electronic structure and the assignments of the absorption bands of steady compounds were discussed. The calculated HOMO and LUMO energies show that charge transfer occurs within the molecules. PMID- 21646045 TI - Enhanced contractile force generation by artificial skeletal muscle tissues using IGF-I gene-engineered myoblast cells. AB - The aim of this study was to investigate whether insulin-like growth factor (IGF) I gene delivery to myoblast cells promotes the contractile force generated by hydrogel-based tissue-engineered skeletal muscles in vitro. Two retroviral vectors allowing doxycycline (Dox)-inducible expression of the IGF-I gene were transduced into mouse myoblast C2C12 cells to evaluate the effects of IGF-I gene expression on these cells. IGF-I gene expression stimulated the proliferation of C2C12 cells, and a significant increase in the growth rate was observed for IGF-I transduced C2C12 cells with Dox addition, designated C2C12/IGF (Dox+) cells. Quantitative morphometric analyses showed that the myotubes induced from C2C12/IGF (Dox+) cells had a larger area and a greater width than control myotubes induced from normal C2C12 cells. Artificial skeletal muscle tissues were prepared from the respective cells using hydrogels composed of type I collagen and Matrigel. Western blot analyses revealed that the C2C12/IGF (Dox+) tissue constructs showed activation of a skeletal muscle hypertrophy marker (Akt) and enhanced expression of muscle-specific markers (myogenin, myosin heavy chain and tropomyosin). Moreover, the creatine kinase activity was increased in the C2C12/IGF (Dox+) tissue constructs. The C2C12/IGF (Dox+) tissue constructs contracted in response to electrical pulses, and generated a significantly higher physical force than the control C2C12 tissue constructs. These findings indicate that IGF-I gene transfer has the potential to yield functional skeletal muscle substitutes that are capable of in vivo restoration of the load-bearing function of injured muscle or acting as in vitro electrically-controlled bio-actuators. PMID- 21646046 TI - Biofilm formation as a function of adhesin, growth medium, substratum and strain type. AB - Biofilm formation is involved in the majority of bacterial infections. Comparing six Escherichia coli and Klebsiella pneumoniae isolates revealed significant differences in biofilm formation depending on the growth medium. Fimbriae are known to be involved in biofilm formation, and type 1, F1C and P fimbriae were seen to influence biofilm formation significantly different depending on strain background, growth media and aeration as well as surface material. Altogether, this report clearly demonstrates that biofilm formation of a given strain is highly dependent on experimental design and that specific mechanisms involved in biofilm formation such as fimbrial expression only play a role under certain environmental conditions. This study underscores the importance of careful selection of experimental conditions when investigating bacterial biofilm formation and to take great precaution/care when comparing results from different biofilm studies. PMID- 21646047 TI - FKBPs: opportunistic modifiers or active players in cancer? PMID- 21646048 TI - Neuroprotective, immunosuppressant and antineoplastic properties of mTOR inhibitors: current and emerging therapeutic options. AB - The acronym mTOR defines a family of serine-threonine protein kinase called mammalian target of rapamycin. The major role of these kinases in the cell is to merge extracellular instructions with information about cellular metabolic resources and to control the rate of anabolic and catabolic processes accordingly. In mammalian cells mTOR is present in two distinct heteromeric protein complexes commonly referred to as mTOR complex 1 (mTORC1) and mTOR complex 2 (mTORC2), involved in the control of a wide variety of cellular processes. It has been recently reported that compounds acting modulating mTOR activity, beside mediating the well recognized processes exploited in the anticancer and immunosuppressant effects, are provided with neuroprotective properties. In fact, mTOR is involved in the mechanism of PI3K/Akt-induced upregulation of glutamate transporter 1, GLT1, that is linked to several neuronal disorders such as stroke, Alzheimer's disease, and amyotrophic lateral sclerosis. Furthermore, in adult brain mTOR is crucial for numerous physiological processes such as synaptic plasticity, learning, memory, and brain control of food uptake. Moreover, the activation of mTOR pathway is involved in neuronal development, dendrite development and spine morphogenesis. PMID- 21646049 TI - Prepuce preserving versus conventional Mathieu urethroplasty for distal hypospadias - a prospective randomized study. AB - OBJECTIVES: To evaluate the impact of prepuce preservation during Mathieu repair for distal hypospadias. METHODS: This prospective randomized comparative study, carried out in Assiut University Hospital, between 2006 and 2009, included 200 patients with distal penile or coronal hypospadias. They underwent either Mathieu repair with prepuce preservation (group I, n = 100) or traditional Mathieu repair with circumcision (group II, n = 100). Follow up was at 2 weeks, and 1 and 3 months postoperatively, which only 153 patients completed (I: n = 86 and II: n = 67). RESULTS: The mean operative time was 64.5 min in group I and 121.7 min in group II (p < 0.001). All patients in group I developed postoperative preputial edema. Patients from group I who had a successful operation underwent circumcision at least 3 months later. Urethrocutaneous fistulae developed in 7 and 6 patients in group I and II respectively (p = 0.967). Fistula closure was done at least 3 months postoperatively, and there was no significant difference in success between the two groups. Two cases of glanular dehiscence were detected (one in each group); the patient from group I had a successful onlay island flap repair. CONCLUSIONS: Preputial preservation during Mathieu repair is a time saving procedure with similar complication rate to traditional repair. Valuable local tissue is preserved to deal with complications that may occur. PMID- 21646051 TI - Magnetic resonance imaging and neuropsychological results from a trial of memantine in Alzheimer's disease. AB - BACKGROUND: This study was designed to assess changes in brain volume and cognitive abilities in memantine-treated patients with Alzheimer's disease (AD) by using an exploratory, single-arm, delayed-start design. METHODS: Cholinesterase inhibitor-treated patients with AD (N = 47; Mini-Mental State Examination score range: 15-23) were enrolled in an observational lead-in period (weeks: 1-24), followed by an open-label period of add-on memantine treatment (weeks: 25-48). The patients underwent magnetic resonance imaging at weeks 0 (baseline), 24 (immediately before memantine initiation), and 48 (endpoint), and a battery of neuropsychological tests at weeks 0, 24, 28, 36, and 48. The primary outcome measure was the annualized rate of change (%) in total brain volume (TBV) between the two study periods. Data were analyzed using paired t-tests. RESULTS: There were no statistically significant differences in the rates of change in TBV, ventricular volume, or left hippocampal volume between the study periods; however, the memantine treatment period was associated with a significantly slower right hippocampal atrophy (-5.5% +/- 12.0% vs -10.8% +/- 7.2%; P = .038). Memantine treatment was also associated with superior performances on the Boston Naming Test (P = .034) and the Trail Making Test, Part B (P = .001), but also with a higher number of errors (i.e., repetitions and intrusions) on the California Verbal Learning Test. Memantine was found to be safe and well tolerated. CONCLUSIONS: In this study, no difference in the rates of TBV change between the two periods was observed; however, memantine treatment was found to be associated with slowing of right hippocampal atrophy, and with improvement on one test of executive functioning as well as a test of confrontation naming ability. Trials using structural magnetic resonance imaging and a delayed-start design may be a feasible option for the assessment of treatments for AD. PMID- 21646050 TI - Developing understanding of the roles of CD1d-restricted T cell subsets in cancer: reversing tumor-induced defects. AB - Invariant natural killer T-cells ('iNKT') are the best-known CD1d-restricted T cells, with recently-defined roles in controlling adaptive immunity. CD1d restricted T-cells can rapidly produce large amounts of Th1 and/or Th2//Treg/Th17 type cytokines, thereby regulating immunity. iNKT can stimulate potent anti-tumor immune responses via production of Th1 cytokines, direct cytotoxicity, and activation of effectors. However, Th2//Treg-type iNKT can inhibit anti-tumor activity. Furthermore, iNKT are decreased and/or reversibly functionally impaired in many advanced cancers. In some cases, CD1d-restricted T-cell cancer defects can be traced to CD1d(+) tumor interactions, since hematopoietic, prostate, and some other tumors can express CD1d. Ligand and IL-12 can reverse iNKT defects and therapeutic opportunities exist in correcting such defects alone and in combination. Early stage clinical trials have shown potential for reconstitution of iNKT IFN-gamma responses and evidence of activity in a subset of patients, with rational new approaches to capitalize on this progress ongoing, as will be discussed here. PMID- 21646052 TI - Altered norepinephrine content and ventricular function in p75NTR-/- mice after myocardial infarction. AB - Cardiac sympathetic neurons stimulate heart rate and the force of contraction through release of norepinephrine. Nerve growth factor modulates sympathetic transmission through activation of TrkA and p75NTR. Nerve growth factor plays an important role in post-infarct sympathetic remodeling. We used mice lacking p75NTR to examine the effect of altered nerve growth factor signaling on sympathetic neuropeptide expression, cardiac norepinephrine, and ventricular function after myocardial infarction. Infarct size was similar in wildtype and p75NTR-/- mice after ischemia-reperfusion surgery. Likewise, mRNAs encoding vasoactive intestinal peptide, galanin, and pituitary adenylate cyclase activating peptides were identical in wildtype and p75NTR-/- cardiac sympathetic neurons, as was expression of the TrkA neurotrophin receptor. Norepinephrine content was elevated in the base of the p75NTR-/- ventricle compared to wildtype, but levels were identical below the site of occlusion. Left ventricular pressure, dP/dt(MAX), and dP/dt(MIN) were measured under isoflurane anesthesia 3 and 7 days after surgery. Ventricular pressure decreased significantly 3 days after infarction, and deficits in dP/dt(MAX) were revealed by stimulating beta receptors with dobutamine and release of endogenous norepinephrine with tyramine. dP/dt(MIN) was not altered by genotype or surgical group. Few differences were observed between genotypes 3 days after surgery, in contrast to low pressure and dP/dt(MAX) previously reported in control p75NTR-/- animals. Seven days after surgery ventricular pressure and dP/dt(MAX) were significantly lower in p75NTR-/- hearts compared to WT hearts. Thus, the lack of p75NTR did not enhance cardiac function after myocardial infarction. PMID- 21646053 TI - Enantiomeric separation of mirtazapine and its metabolite in rat plasma by reverse polar ionic liquid chromatography using fluorescence and polarimetric detectors connected in series. AB - A simple and rapid reverse polar ionic LC method was developed and validated for simultaneous separation and determination of mirtazapine, an antidepressant drug, and its main metabolite N-desmethyl mirtazapine using fluorescence and polarimetric detectors connected in series. The chromatographic separation was achieved on Chirobiotic V column packed with vancomycin as a stationary phase in an isocratic mode of elution of methanol:glacial acetic acid:anhydrous triethyl amine (100:0.2:0.1, v/v/v) as a mobile phase. The compounds were detected by their excitation at 290nm and emission at 370nm using fluorescence detector while the optical rotation (+/-) of the enantiomers was identified by polarimetric detector. The analytes were extracted from rat plasma by precipitation of proteins and the average yield was 88-111% for mirtazapine and 85-123% for N desmethyl mirtazapine. The method was linear over the concentration range of 20 5000ng/mL. The method was successfully applied on rat plasma spiked with the enantiomers of mirtazapine and N-desmethyl mirtazapine. PMID- 21646055 TI - Correction of the severely inverted nipple: areola- based dermoglandular rhomboid advancement. AB - Inverted nipple is a relatively common aesthetic problem presenting to a plastic surgeon. Along with the functional problems; recurrent inflammation/infection and an inability to nurse, most patients seek intervention because of the abnormal appearance. Many different surgical techniques have been described, suggesting that no one technique is universally successful. Most techniques employ, either individually or in combination, methods to tighten the nipple base or the use of areolar dermal flaps to support the nipple. We propose two modifications to the dermal flap technique. Firstly, the rhomboid dermal flaps are designed solely on the areola, the bases directed peripherally, improving vasularity and allowing true medial advancement. Secondly, the lactiferous ducts and tethering fibrous bands are released in a conical fashion allowing closure with a vertical V-Y advancement. This gives better projection and support to the nipple proper with medial recruitment of glandular tissue closing the dead space created. This technique has been used successfully by the senior author in 20 patients over a 16 year period, with a high rate of preserved nipple evertion and patient satisfaction. The technique is simple, reliable and provides sustained results over long term follow-up for the correction of the more severely inverted nipple. PMID- 21646054 TI - Centrosome clustering and chromosomal (in)stability: a matter of life and death. AB - Centrosome abnormalities occur commonly in cancer, and contribute to chromosomal instability and tumorigenesis. New evidence on a phylogenetically conserved mechanism termed 'centrosomal clustering' provides exciting insights into how cells with supernumerary centrosomes adapt to avoid lethal multipolar divisions. Here, we highlight the emerging molecular basis of centrosome clustering, and its impact on asymmetric divisions of stem cells, chromosomal (in)stability and malignant transformation. Finally, pharmacological inhibition of centrosome clustering promises to selectively target tumor cells. PMID- 21646056 TI - MI sensor-aided screening system for assessing swallowing dysfunction: application to the repetitive saliva-swallowing test. AB - PURPOSE: Assessment of swallowing dysfunction, particularly the risk of aspiration, is extremely important to clinicians because it provides crucial information for preventing morbidity and mortality from dysphagia. The purpose of this study was to describe a magneto-impedance sensor-aided screening system (MISS) for assessing swallowing function and clarify its effectiveness with the repetitive saliva-swallowing test (RSST). METHODS: An MI sensor attached to the skin over the sternum detected the distance from a magnet attached to the skin over the thyroid cartilage during swallowing as the change of magnetic fields. The MISS was validated by videofluoroscopic and videoendoscopic observations. Further, the swallowing behavior of 93 individuals was assessed by using the MISS and then analyzed with the RSST. Swallowing behavior can be recorded and investigated objectively by signal inspection in the MISS compared with the conventional methods. The MISS is a simple and straightforward method for recording and safe because of dry swallow. The MISS system was validated by using simultaneous recordings with videofluoroscopic or videoendoscopic examinations, and compared with the conventional RSST method. CONCLUSIONS: The MISS combined with the RSST is an effective screening test for swallowing function. PMID- 21646057 TI - EuroPCR 2011: the ongoing challenge of initiatives and innovations. PMID- 21646058 TI - Peripheral interventions: how long will they remain a missed opportunity? PMID- 21646059 TI - Identification of high-risk aortic valve patients. PMID- 21646060 TI - Prevalence and prognostic implications of baseline anaemia in patients undergoing transcatheter aortic valve implantation. AB - AIMS: TAVI is a minimally invasive alternative to surgical aortic valve replacement or medical therapy in patients with a high or prohibitive operative risk. The clinical significance of baseline anemia and prognostic implications in this patient cohort are unknown. We sought to evaluate the prevalence and prognostic implications of baseline anaemia in patients undergoing transcatheter aortic valve implantation (TAVI) at our institution. METHODS AND RESULTS: One hundred and eighteen consecutive patients who underwent TAVI with the Medtronic Corevalve System (Medtronic Corp., Minneapolis, MN, USA) were included in the analysis. Clinical and biochemical data were prospectively collected before, during and after the procedure. Clinical follow-up was set at one month, one year and yearly thereafter. Anaemia was defined as a haemoglobin level <13 g/dL in men and <12 g/dL in women. Mortality was confirmed by consultation of the civil registry. The prevalence of baseline anaemia was 49%. Anaemic patients undergoing TAVI required more RBC transfusions (3.3 +/- 3.1 versus 1.5 +/- 2.3; p<0.001) and more frequently experienced a prolonged index hospitalisation exceeding two weeks. For patients with at least 1-year follow up (N=74), mortality at 30 days was no different; however 1-year mortality was significantly higher in the anaemic cohort (44 versus 15%, p=0.006). In a multivariable analysis, baseline anaemia emerged as an independent predictor of 1- year mortality (HR 2.10 [1.06 4.18]). CONCLUSIONS: In our series, baseline anaemia is common in patients undergoing TAVI, forecasts a need for more red blood cell transfusions and is associated with increased 1-year mortality. PMID- 21646061 TI - Multicentre experience with the BridgePoint devices to facilitate recanalisation of chronic total coronary occlusions through controlled subintimal re-entry. AB - AIMS: The major challenge for the interventional treatment of chronic total coronary occlusion (CTO) is a low primary success rate. A common problem is the passage of the recanalisation wire into a subintimal position. New devices, which were evaluated in the first multicentre study in CTOs resistant to a conventional wire approach, may help to facilitate a controlled re-entry into the true lumen. The aim of this study was to assess the safety and efficacy of this approach, with successful true lumen distal wire passage as the primary endpoint. METHODS AND RESULTS: Forty-two patients were enrolled in four centres with high expertise in PCI for CTOs. All CTOs were of at least three months duration, and were initially attempted with dedicated recanalisation wires. After failure to pass or creation of a subintimal dissection, the BridgePoint devices were applied, consisting of a ball-tipped catheter (CrossBoss) to pass the proximal occlusion cap, and a flat-shaped balloon catheter (Stingray catheter) to be inflated within the subintimal space to guide the re-entry into the true vessel lumen with a special wire (Stingray guidewire). The primary endpoint was met in 67% of all patients. A higher success rate seemed to be possible when all devices were used in sequenced beginning with the CrossBoss, and in the case of a subintimal passage, followed by the Stingray. True lumen re-entry failed because of the loss of distally contrast filling and thus loss of a target for re-entry, and by a failure to advance the Stingray balloon far enough distal and parallel to the distal lumen. There were no severe device related complications. CONCLUSIONS: In patients with complex CTOs referred to dedicated centres with high experience in CTOs, these results demonstrate the potential of a guided re-entry from a subintimal wire position by use of the BridgePoint devices. PMID- 21646062 TI - Clinical outcome of percutaneous treatment of in-stent restenosis with drug eluting stents: results from the first phase of the prospective multicentre German DES.DE registry. AB - AIMS: Treatment of in-stent restenosis (ISR) was historically considered the Achilles heel of percutaneous coronary intervention (PCI) and has been associated with worse clinical outcome than PCI of de novo lesions. However, comparative data on ISR and de novo lesions using drug-eluting stents (DES) are scarce. Therefore, we aimed to assess the impact of ISR on procedural and long-term outcome in patients treated with DES. METHODS AND RESULTS: We analysed data from 5,144 patients enrolled in the prospective multicentre German Drug-Eluting Stent Registry (DES.DE). The registry included 872 patients (17%) treated for ISR with follow-up data (median 12.4 months) available for 817 patients (94%). Of the ISR patients, 37.1% (n=323) presented with acute coronary syndromes. In total, 1,027 DES were used (528 sirolimus-eluting stents and 499 paclitaxel-eluting stents), with successful implantation in 97.7% of patients. In the ISR cohort, myocardial infarction (MI) during hospitalisation was observed in 1.6% of patients (n=14) and in-hospital mortality was only 0.3% (n=3). Major adverse cardiac and cerebrovascular events (MACCE) rate at follow-up (defined as a composite of death, MI and stroke) was 8.7% (n=71) versus 8.2% (n=325) in patients treated for de novo lesions (p=0.63). Target vessel revascularisation (TVR) rate was 12.7% (n=100), numerically higher than in patients with de novo lesions (10.5%, p=0.07). Ten patients (1.3%) suffered from ARC definite stent thrombosis versus 0.7% observed in patients with de novo lesions (p=0.13). After adjustment for differences in baseline characteristics, TVR rates were statistically higher in the ISR cohort (OR 1.27, 95%CI 1.01-1.61, p=0.04), while MACCE rates remained comparable (OR 1.10, 95%CI 0.83-1.44, p=0.51). The type of stent used (sirolimus vs. paclitaxel-eluting stent) did not impact the rate of MACCE, TVR or definite stent thrombosis at one year. CONCLUSIONS: Results from this large prospective multicentre registry confirm that treatment of ISR with DES is effective and safe, with similar procedural outcome but slightly higher revascularisation rates at one year compared to patients treated for de novo lesions, with no differences in outcome between sirolimus- and paclitaxel-eluting stents. PMID- 21646063 TI - Comparison of outcome after patent foramen ovale closure in older versus younger patients. AB - AIMS: Percutaneous patent foramen ovale (PFO) closure seems to be effective for secondary prevention of cryptogenic stroke in patients younger than 55 years of age. The efficacy in older patients remains uncertain. We compared the efficacy of PFO closure between patients younger and older than 55 years. METHODS AND RESULTS: All 335 patients (mean age 50.2 +/- 12.6 years; 205 men) with cryptogenic thromboembolism who underwent PFO closure in our centres between 1998 and 2008 were included. Mean follow-up period was 4.2 +/- 1.9 years in the elderly (n=120) and 3.8 +/- 2.4 years in the younger patients (n=215) (p=0.15). Prevalence of hypertension, diabetes, hyperlipidaemia and coronary and peripheral artery disease was higher in the elderly (p<0.05 for all). Re-occurrence of stroke or TIA was higher in the elderly compared to the younger (annual event rate 2.4% versus 0.6%; log rank, p=0.005). Re-occurrence of stroke alone was higher in the elderly (annual event rate 1.2% versus 0.1%; log rank, p=0.01). Multivariate analysis showed that an age of >55 years was an independent predictor of recurrent stroke or TIA (HR 3.2, p=0.03). CONCLUSIONS: Percutaneous PFO closure appears to be effective for secondary prevention of cryptogenic stroke in younger patients but seems to be related with less beneficial outcome in elderly. Randomised controlled trials are needed to confirm our findings. PMID- 21646064 TI - Clinical and procedural evaluation of the Nile Croco(r) dedicated stent for bifurcations: a single centre experience with the first 151 consecutive non selected patients. AB - AIMS: To evaluate the feasibility and safety of the Nile Croco(r) coronary bifurcation stent system (Minvasys, Gennevilliers, France). METHODS AND RESULTS: The primary endpoint was to assess the acute device success and angiographic success with the use of the Nile Croco(r) stent system. Secondary endpoints included in-hospital and six month major cardiac events (MACE).There were 151 consecutive patients enrolled in the Nile Croco Study at Vall Hebron Hospital. The Nile Croco(r) stent was successfully implanted in 144 patients (95.4%) and final angiographic success was obtained in 100% of the patients. 138 out of the 151 (91%) patients included have accomplished the six month follow-up. There was one in-hospital MACE in the 151 recruited patients. The MACE rate at six months in the 138 patients with follow-up was 14% and the ischaemia-driven TLR rate was 7.2 %. CONCLUSIONS: The results of our Nile Croco(r) Study are the first to demonstrate the safety and high performance of this dedicated stent system for the treatment of bifurcation lesions. The device can be successfully implanted in more than 95% of all cases, with a high procedural success rate and low in hospital and six month MACE rates. PMID- 21646065 TI - Correlation between fractional flow reserve and intravascular ultrasound lumen area in intermediate coronary artery stenosis. AB - AIMS: Fractional flow reserve (FFR) of <0.8 or 0.75 is currently used to guide revascularisation in lesions with intermediate coronary stenosis. We assessed whether there is an intravascular ultrasound (IVUS) measurement that can reliably be used to predict when patients should undergo intervention. METHODS AND RESULTS: The analysis included 92 intermediate lesions (84 patients) located in vessel diameters >2.5 mm. Positive FFR was considered present at <0.8 and 0.75. IVUS minimum lumen area (MLA) was correlated to the FFR findings in intermediate lesions with 40-70% stenosis. The mean FFR value was 0.89 +/- 0.08. Twenty-four patients (26.1%) had FFR <0.8; 17 (18.5%) <0.75. Positive correlations between FFR and IVUS measurements included MLA (r = 0.34, p<0.001), minimum lumen diameter (MLD) (r=0.31, p=0.004), lesion length (r=-0.5, p<0.001), and area stenosis (r=-0.31, p=0.01). There was no significant correlation between FFR and quantitative coronary angiography in MLD (r=0.19, p=0.06), diameter stenosis (r=0.08, p=0.4), or lesion length (r=-0.14, p=0.17). A receiver operating characteristic curve identified MLA <2.8 mm2 (sensitivity 79.7%, specificity 80.3%) as the best threshold value for FFR <0.75; and MLA <3.2 mm2 as best for FFR <0.8 (sensitivity 69.2%, specificity 68.3%). CONCLUSIONS: Anatomic measurements of intermediate coronary lesions obtained by IVUS show a moderate correlation to FFR values, although they differ according to vessel size. IVUS MLA may be used as an alternative to FFR when assessing the need for intervention in intermediate coronary lesion. Vessel size, however, should always be taken into account. PMID- 21646066 TI - The Angio-SealTM femoral closure device allows immediate ambulation after coronary angiography and percutaneous coronary intervention. AB - AIMS: To test the safety of immediate mobilisation of patients undergoing coronary angiography and percutaneous coronary intervention (PCI) closed with Angio-SealTM -a femoral vascular closure device. METHODS AND RESULTS: First, a randomised controlled trial of immediate mobilisation vs. delayed ambulation was performed followed by a prospective validation registry to test the obtained results in a real-world situation. The randomised trial comprised 300 patients; the validation registry comprised 1,097 patients. Primary endpoints were complications defined as: small haematoma <5 cm and/or minor bleeding/oozing from the puncture site, haematomas >= 5 cm, bleeding needing transfusion, bleeding needing surgical attention, pseudoaneurysm and vasovagal reaction. In the randomised trial, overall complications were similar in both groups (16.0%vs.18.8%; p=0.53). Small haematomas/small bleedings/oozing were the most frequent (12.2% vs.15.3; p=0.44). There were no bleedings needing transfusion or surgical attention, and no pseudoaneurysms occurred. The prospective registry showed similar results. In the standard-care cohort, complications were similar to those in the implementation cohort (9.6% vs.11.3%; p=0.41), mainly consisting of small haematomas/minor bleedings/oozing (6.1% vs.7.3%; p=0.49). No bleedings needed transfusion or surgical attention. Pseudoaneurysms occurred in 1 (0.34%) vs. 3 (0.37%; p=0.94) and vasovagal reactions in three (1.0%) vs. four (0.5%; p=0.33) patients. It was possible to mobilise 87% of patients in the implementation cohort. CONCLUSIONS: In patients undergoing coronary angiography or PCI, the use of immediate mobilisation after Angio-SealTM deployment is safe. With routine use of a femoral vascular closure device, approximately 87% of patients are suitable for immediate mobilisation. PMID- 21646067 TI - Cardio-respiratory exercise testing early after the use of the Angio-Seal system for arterial puncture site closure after coronary angioplasty. AB - AIMS: The vascular closure device (VCD) Angio-Seal is an easy-to-use system for the closure of arterial puncture sites after percutaneous coronary intervention (PCI), and allows for early mobilisation of the patient. However, little data are available about exercising early after the use of VCD's in PCI patients. METHODS AND RESULTS: A total of 230 consecutive patients were screened. Of these, 45 (20%) were excluded due to the inability to perform exercise testing, or anatomical conditions which prevented the insertion of a VCD. The 185 remaining patients (139 male, mean age 68 +/- 12 years) received Angio-Seal after PCI. After four hours, 30 patients (16%) showed a small local haematoma, 11 patients (6%) complained about minor -and one patient (0.5%) about strong- groin pain. There were no major bleeding complications, six pseudo-aneurysmata, and one arterio-venous fistula. Overall, nine patients (6%) showed moderate to severe groin problems. Patients without major complications underwent bicycle cardiopulmonary exercise testing the subsequent day. Exercise testing was performed up to 136 +/- 60 W in 176 patients (94%). Maximum workload was 104 +/- 33 W, peak oxygen consumption 17.6 +/- 5.1 ml/min/kg, and oxygen consumption at the anaerobic threshold 15.4 +/- 4.2 ml/min/kg. After exercise testing there were no cardiovascular complications noted. CONCLUSIONS: In patients receiving VCD after PCI, exercise testing above the anaerobic threshold was feasible after Angio-Seal deployment in those patients with no complications after the use of the device. PMID- 21646068 TI - A simple nomogram for early prediction of myocardial reperfusion after pre hospital thrombolysis. AB - AIMS: To discriminate early ST-segment elevation myocardial infarction (STEMI) presenters at a high probability of successful pre-hospital thrombolysis (PHT) using a simple nomogram based on independent predictors of complete ST resolution. METHODS AND RESULTS: OPTIMAL was an observational, prospective study undertaken at 79 medical centres in France in patients with STEMI undergoing pre hospital thrombolysis (PHT) within six hours of symptom onset and coronary angiography within six hours of thrombolysis. The baseline and pre-coronary angiography ECGs of 800 patients were analysed. The main outcome measure was ST segment resolution >= 70%. ST resolution was associated with a significant reduction in mortality (1.8% vs. 4.3%; p=0.05). After multivariable logistic regression analysis, five independent predictors of successful myocardial reperfusion were identified: <= 1 h between pain onset and thrombolysis (odds ratio [OR] 1.76, 95% confidence interval [CI] 1.18-2.62); body mass index (BMI) <30 kg/m2 (OR 1.74, CI 1.12-2.72); current/previous smoking (OR 1.71, CI 1.21 2.43); non-anterior infarct territory (OR 1.75, CI 1.27-2.41); and maximum amplitude of ST elevation <3 mm (OR 1.83, CI 1.32-2.54). The C-statistic of the model was 0.71 (95% CI 0.67-0.74). Using these five independent factors, a simple nomogram was developed to assess the probability of ST resolution after PHT. This nomogram allows discrimination of probabilities ranging from 13 to 72%. CONCLUSIONS: This simple nomogram can predict the probability of successful myocardial reperfusion after thrombolysis. This may be useful in the triage of STEMI presenters. PMID- 21646069 TI - Major stent deformation/pseudofracture of 7 Crown Endeavor/Micro Driver stent platform: incidence and causative factors. AB - AIMS: This is the initial report of stent deformation/pseudofracture of the 7 crown Endeavor/Micro Driver stent platform (2.25-2.75 mm), whereby the post dilation balloon catches and causes major stent deformation angiographically appearing as a large stent fracture. We sought to determine frequency and cause. METHODS AND RESULTS: Analysis of 1,000 consecutive Endeavor/Micro Driver stents (7 crown) deployed at our institution. Bench testing was also performed by deploying 10 stents in 2.5 mm tubing and then attempting re-crossing with non compliant balloons. CLINICAL RESULTS: There were 14 cases of major stent deformation/pseudofracture representing an incidence of 1.4% or 1.8% of 775 stents that were post-dilated. Of the 14 deformed stents, re-stenting was required in nine and MACE occurred in five (36%.). Benchtop results: Balloon "catch" was reproducible at an initial frequency of 9% and affected three of the 10 stents. With the use of provocative measures stent deformation occurred in 50% of test stents. CONCLUSIONS: Stent deformation/pseudofracture of the 7 crown Endeavor/Micro Driver platform occurred in 1.8% of cases that were post-dilated. Of these, 36% experience MACE. Deformation can be reproduced on the benchtop. Operators should be aware of the complication, and develop strategies to deal with it. PMID- 21646070 TI - Looking back into the future: desirudin in acute coronary syndromes and coronary stenting. AB - Although percutaneous coronary intervention (PCI) is a highly effective modality for the management of acute coronary syndromes, it can potentiate the existing prothrombotic state around lesion areas and lead to ischaemic complications. Adjunctive pharmacologic treatment with heparin reduces the risk of ischaemic events, but the utility of heparin is limited by its unpredictable pharmacodynamic effects and its inability to modulate fibrin-bound thrombin. Additionally, a potential risk of heparin-induced thrombocytopenia is associated with heparin use. Direct thrombin inhibitors (DTIs) have emerged as potential alternatives to heparin in patients undergoing PCI. Bivalirudin is a DTI indicated for use in PCI. Results from various studies have suggested clinical benefit associated with the use of bivalirudin, driven primarily by the reduction in bleeding risks compared with the standard treatment regimens. Of concern, however, is a significant increase in acute stent thrombosis with bivalirudin monotherapy compared with heparin plus GPIIb/IIIa inhibitors following primary PCI for ST-segment elevation myocardial infarction (STEMI). Desirudin is a highly potent DTI with greater binding affinity than bivalirudin for thrombin. This report provides a comparative overview of the pharmacology and clinical utility of desirudin and bivalirudin in the setting of PCI. PMID- 21646071 TI - Coronary dissection healing patterns: from complete resolution to restenosis, insights from optical coherence tomography. AB - A 68-year-old patient presented a long, type C coronary dissection after balloon angioplasty of a focal lesion in a distal marginal branch. Due to the small vessel size the dissection was left untreated. At angiographic follow-up the dissection had completely disappeared, but a localised restenosis was found in the proximal vessel. Optical coherence tomography (OCT) confirmed the complete healing of the dissection and revealed a normal vessel wall (restitutio ad integrum). This technique also provided unique insights into the underlying substrate of the proximal restenosis unravelling a complicated plaque with associated thrombus. PMID- 21646072 TI - Moxy(r) drug-coated balloon: a novel device for the treatment of coronary and peripheral vascular disease. PMID- 21646073 TI - How should I treat a huge intra-mural haematoma of the ascending aorta after coronary angioplasty? PMID- 21646074 TI - Tools & Techniques: CTO--the retrograde approach. PMID- 21646075 TI - Healing of a coronary artery dissection detected by intravascular ultrasound and optical coherence tomography. PMID- 21646076 TI - [Contribution of FDG-PET in the management of pediatric sarcomas in 2011]. AB - FDG-PET has boomed in recent years for diagnosis, staging and the search for recurrence of a large number of tumors. This is particularly true for soft tissue sarcomas and musculoskeletal sarcomas, for which the first publications on the potential role of FDG-PET dating back to the early 1990s. The majority of published studies on adult sarcomas confer, possibly a mixed population. Studies dedicated to pediatrics population are much rarer. The "Standards, Options and Recommendations" of the French Federation of Anticancer Centers published in 2003 on "The use of FDG-PET in oncology" and make recommendations and expert advices as part sarcomas of adult patients. After a first part dedicated to the particular interpretation of FDG PET in children, the purpose of this paper is to review the potential contribution of this exam in the treatment of pediatric sarcomas. PMID- 21646077 TI - Stomatal architecture and evolution in basal angiosperms. AB - Stomatal architecture-the number, form, and arrangement of specialized epidermal cells associated with stomatal guard cells-of 46 species of basal angiosperms representing all ANITA grade families and Chloranthaceae was investigated. Leaf clearings and cuticular preparations were examined with light microscopy, and a sample of 100 stomata from each specimen was coded for stomatal type and five other characters contributing to stomatal architecture. New stomatal types were defined, and many species were examined and illustrated for the first time. Character evolution was examined in light of the ANITA hypothesis using MacClade software. Analysis of character evolution, along with other evidence from this study and evidence from the literature on fossil angiosperms and other seed plant lineages, suggests that the ancestral condition of angiosperms can be described as anomo-stephanocytic, a system in which complexes lacking subdidiaries (anomocytic) intergrade with those having weakly differentiated subsidiaries arranged in a rosette (stephanocytic). From this ancestral condition, tangential divisions of contact cells led to the profusion of different types seen in early fossil angiosperms and Amborellaceae, Austrobaileyales, and derived Chloranthaceae, while the state in Nymphaeales is little modified. Formation of new, derived types by tangential division appears to be a recurrent theme in seed plant evolution. PMID- 21646078 TI - Correlated evolution of leaf shape and trichomes in Begonia dregei (Begoniaceae). AB - Structural features of leaves, including size, shape, and surfaces, vary greatly throughout the plant kingdom. In both functional and phylogenetic analyses of leaves, the various morphological aspects are often considered independently of each other, although it is likely that many combinations of features do not occur at random due to either functional constraint or genetic correlation. The distribution of variation in leaf morphology in the highly variable Begonia dregei species complex was examined in natural populations and in F(2) offspring from a cross between plants from two populations. Leaf shape was quantified using several morphometric measures, and trichomes on leaves were counted and measured. Correlations between leaf shape and the numbers and size of trichomes were examined. There were significant correlations between the shapes of leaves and the presence, number, and size of trichomes among populations and in hybrid plants. Deeply incised leaves had larger numbers of longer trichomes at the sinuses. Higher numbers of trichomes on upper leaf surfaces occurred together with trichomes at the petiole and on the abaxial surface. The potential for independent evolution of leaf shape and trichomes in this group is limited. Hypotheses to explain the correlated development of leaf shape and trichomes are discussed. PMID- 21646079 TI - Chemistry and geographic variation of floral scent in Yucca filamentosa (Agavaceae). AB - We identified volatiles from the floral headspace of Yucca filamentosa using gas chromatography and mass spectrometry and analyzed floral scent composition and variation among populations pollinated by different yucca moth species. Twenty one scent compounds were repeatedly identified and most could be categorized into two major classes: (1) homoterpenes derived from the sesquiterpene alcohol nerolidol and (2) long chain aliphatic hydrocarbons. Two biosynthetic pathways are thus responsible for the majority of floral volatiles in Y. filamentosa. The homoterpene E-4,8-dimethylnona-1,3,7-triene, which is released systemically by higher plants upon herbivory, was the most abundant compound. Two di-oxygenated compounds not previously reported as floral compounds also were detected. No differentiation in floral scent was observed between populations pollinated by different yucca moths, nor was there any correlation between chemical distance and geographic distance among populations. The total release rate of volatiles differed significantly among populations, but not between populations with different pollinators. The combination of unique compounds and low variation in the fragrance blend may reflect highly selective attraction of obligate pollinators to flowers. The observed lack of differentiation in floral scent can putatively explain high moth-mediated gene flow among sites, but it does not explain conservation of odor composition across populations with different pollinators. PMID- 21646080 TI - Influence of plant ontogeny on compensation to leaf damage. AB - Ontogenetic changes in architecture, carbohydrate reserves, and resource allocation can constrain the ability of plants to compensate for herbivore damage. To evaluate ontogenetic changes in compensation, saplings and reproductive individuals of the tropical tree Casearia nitida were subjected to three levels of defoliation (0, 25, and 75% leaf area removed) and regrowth was quantified. The impact of defoliation on fruit production was evaluated in reproductive trees. In addition, the influence of defoliation on carbohydrate reserves and on the production of phenolic compounds was assessed. Plants at both stages were able to compensate for 25% leaf area loss, but only saplings were able to compensate at the 75% defoliation level. Negative impacts of defoliation on reproductive trees were also suggested by their tendency to produce fewer fruits when defoliated. The concentration of nonstructural carbohydrates decreased with damage in saplings but not in reproductive trees, suggesting an ontogenetic stage-dependent impact of defoliation on carbohydrate reserves. The concentration of phenolic compounds in leaves decreased with increasing leaf damage in both ontogenetic stages. This suggests a resource based trade-off between defense and compensation. The results from this study suggest that ontogeny needs to be considered when assessing plant responses to herbivore damage. PMID- 21646081 TI - Effects of herbivory and inbreeding on the pollinators and mating system of Mimulus guttatus (Phrymaceae). AB - Most models of mating system evolution predict mixed mating to be unstable, although it is commonly reported from nature. Ecological interactions with mutualistic pollinators can help account for this discrepancy, but antagonists such as herbivores are also likely to play a role. In addition, inbreeding can alter ecological interactions and directly affect selfing rates, which may also contribute to maintaining mating system variation. We explored herbivore and inbreeding effects on pollinator behavior and selfing rates in Mimulus guttatus. First, individual spittlebug (Philaenus spumarius) herbivores were applied to native plants in two populations. Spittlebugs reduced flower size, increased anther-stigma distance, and increased selfing rates. A second experiment factorially crossed spittlebug treatment with inbreeding history (self- vs. cross fertilized), using potted plants in arrays. Spittlebugs did not affect pollinator behavior, but they reduced flower size and nearly doubled the selfing rate. Inbreeding reduced the frequency of pollinator visits and increased flower handling time, and this may be the first report that inbreeding affects pollinator behavior. Selfing rates of inbred plants were reduced by one half, which may reflect early inbreeding depression or altered pollinator behavior. The contrasting effects of herbivory and inbreeding on selfing rates may help maintain mating system variation in M. guttatus. PMID- 21646082 TI - Paleoactaea gen. nov. (Ranunculaceae) fruits from the Paleogene of North Dakota and the London Clay. AB - Paleoactea nagelii Pigg & DeVore gen. et sp. nov. is described for a small, ovoid ranunculaceous fossil fruit from the Late Paleocene Almont and Beicegel Creek floras of North Dakota, USA. Fruits are 5-7 mm wide, 4.5-6 mm high, 10-13 mm long, and bilaterally symmetrical, containing 10-17 seeds attached on the upper margin in 2-3 rows. A distinctive honeycomb pattern is formed where adjacent seeds with prominent palisade outer cell layers abut. Seeds are flattened, ovoid, and triangular. To the inside of the palisade cells, the seed coat has a region of isodiametric cells that become more tangentially elongate toward the center. The embryo cavity is replaced by an opaline cast. This fruit bears a striking resemblance to extant Actaea, the baneberry (Ranunculaceae), an herbaceous spring wildflower of North Temperate regions. A second species, Paleoactaea bowerbanki (Reid & Chandler) Pigg & DeVore nov. comb., is recognized from the Early Eocene London Clay flora, based on a single fruit. This fruit shares most of the organization and structure of P. nagelii but is larger and has a thicker pericarp. This study documents a rare Paleocene occurrence of a member of the buttercup family, a family that is today primarily herbaceous, and demonstrates a North Atlantic connection for an Actaea-like genus in the Paleogene. PMID- 21646083 TI - (1->3),(1->4)-{beta}-d-Glucans in the cell walls of the Poales (sensu lato): an immunogold labeling study using a monoclonal antibody. AB - (1->3),(1->4)-beta-Glucans had previously been detected in nonlignified cell wall preparations of only the Poaceae and five other families in the graminoid clade of the Poales (s.l.). Cell walls of vegetative organs of 12 species in nine families of the Poales (s.l.) were examined by immunogold labeling using a monoclonal antibody to (1->3),(1->4)-beta-glucans. Three types of wall-labeling patterns were identified depending on the density of labeling of the nonlignified walls of epidermal and parenchyma cells and the lignified walls of sclerenchyma fibers and xylem tracheary elements: type 1 in Poaceae and Flagellariaceae, type 2 in Restionaceae and Xyridaceae, and type 3 in Cyperaceae and Juncaceae. Type 1 had the heaviest labeling of nonlignified walls and type 2 the heaviest labeling of lignified walls. Type 3 had the least wall labeling, with only very light labeling of nonlignified and lignified walls. No labeling was found over walls of Typhaceae, Sparganiaceae, or Bromeliaceae. The results are discussed in relation to Poales phylogeny. PMID- 21646084 TI - Response of root branching to abscisic acid is correlated with nodule formation both in legumes and nonlegumes. AB - Legumes are unique among higher plants in forming a symbiosis with Rhizobium. Phylogenetic studies indicate this symbiosis may have evolved as many as three times within the Fabaceae; alternatively, a predisposition for nodulation evolved early in the history of the legume lineage. We have identified a physiological trait-increased lateral root formation in response to abscisic acid (ABA)- that marks all nodulating and non-nodulating legume species in our study set with the exception of Chamaecrista fasciculata and Cercis occidentalis. In contrast, nonlegume species tested decrease lateral root formation in response to ABA. Cercis is not a descendant of any common ancestor hypothesized to have evolved Rhizobium nodulation and has an intermediate response to ABA, partway between that of nonlegumes and legumes. We suggest that acquisition of altered responsiveness of roots to ABA is coincident with the appearance of a predisposition for nodulation within the legumes, followed by a loss in Chamaecrista. In addition, we demonstrate that altered ABA responsiveness of lateral root formation characterizes roots of the actinorhizal nodulator, Casuarina glauca, but not the closely related, nonactinorhizal species, Betula papyrifera. Thus our data provide evidence for a physiological root trait associated with nodulation both in legumes and in an actinorhizal plant. PMID- 21646085 TI - Genetic variation and structure in the expanding moss Pogonatum dentatum (Polytrichaceae) in its area of origin and in a recently colonized area. AB - Genetic variation in the expanding moss species Pogonatum dentatum was studied using intersimple sequence repeat (ISSR) markers. The genetic consequences of range expansion were studied by comparing source populations in a mountain area with populations from a recently colonized lowland area in Sweden. Indices of genetic variation show slightly lower number of alleles per locus in the lowlands and a similar gene diversity in both areas. Three of four lowland populations had evidence of a recently passed bottleneck. Considerably higher haplotype diversity was found in the recently colonized lowlands compared to source populations in the mountains. Patterns of allelic diversity suggest that P. dentatum experiences loss of genetic variation through founder effects and genetic drift when expanding its distribution range. Higher haplotypic diversity, less linkage disequilibrium, and fewer compatible loci indicate that sexual recombination is relatively more important in the lowlands compared to the mountains. A likely explanation is higher success of establishment from spores in the lowlands, while clonal propagation predominates in the mountains. Less genetic differentiation among lowland populations indicates more gene flow in the lowland area, involving more spores and/or fragments moving among populations. PMID- 21646086 TI - Outbreeding, seedling establishment, and maladaptation in natural and reintroduced populations of rare and common Silene douglasii (Caryophyllaceae). AB - Reintroductions are increasingly used to enhance declining populations, yet comparative data for critical germination and establishment phases are seldom available for both rare and common herbaceous perennials. After introducing a total of >1800 seeds, we compared experimentally manipulated and natural populations of widespread Silene douglasii var. douglasii relative to rare S. douglasii var. oraria, known in only three coastal headlands. Despite equivalent ex situ germination, oraria field plots produced significantly fewer juveniles than douglasii plots indicating that seedling survival limits plant establishment. We also evaluated transplant vs. seed reintroductions as restoration tools, the effect of inbreeding on fitness, and the potential importance of buried seed pools. Germination declined rapidly for seeds over 1-2 years old, and only 2.2% of newly collected seeds of oraria survived as seedlings. Transplant survival over 5 years was greatest for outbred progeny; furthermore, 75% of the new seedlings emerged near outbred progeny from the original reintroduction. Despite similar ovule numbers and pollinator visitation, transplants exhibited 49-179% maladaptation in the formerly grazed site, with significantly lower fruit and seed set than adults in more diverse natural populations. This study experimentally identifies several key factors affecting plant reintroductions, facilitating effective development of large-scale reintroduction strategies for native perennials. PMID- 21646087 TI - Vernalization sensitivity in Arabidopsis thaliana (Brassicaceae): the effects of latitude and FLC variation. AB - Latitudinal variation in climate is predicted to select for latitudinal differentiation in sensitivity to the environmental cues that signal plants to flower at the appropriate time for a given climate. In Arabidopsis thaliana, flowering is promoted by exposure to cold temperatures (vernalization), and several vernalization pathway loci are known. To test whether natural variation in vernalization sensitivity could account for a previously observed latitudinal cline in flowering time in A. thaliana, we exposed 21 European accessions to 0, 10, 20, or 30 d of vernalization and observed leaf number at flowering under short days in a growth chamber. We observed a significant latitudinal cline in vernalization sensitivity: southern accessions were more sensitive to vernalization than northern accessions. In addition, accessions that were late flowering in the absence of vernalization were more sensitive to vernalization cues. Allelic variation at the flowering time regulatory gene FLC was not associated with mean vernalization sensitivity, but one allele class exhibited greater variance in vernalization sensitivity. PMID- 21646088 TI - Sexes show differential tolerance to spittlebug damage and consequences of damage for multi-species interactions. AB - Antagonists can play a role in sexual system evolution if tolerance or resistance is sex-dependent. Our understanding of this role will be enhanced by consideration of the effects of antagonists on other plant-animal interactions. This study determined whether the sex morphs of a gynodioecious Fragaria virginiana differ in their susceptibility and response to damage by spittlebugs and whether damage altered pollinator attraction traits or interactions with other antagonists. Tolerance, but not resistance, to spittlebugs differed between the sexes. Generally, spittlebugs were more damaging to hermaphrodites than females, a finding in accord with the hypothesis that the pollen-bearing morph is less tolerant of source-damage than the pollen-sterile morph when damage is incurred during flowering. In both sex morphs, spittlebugs reduced inflorescence height, increased petal size, but did not affect the number of open flowers per day, suggesting that the net effect of damage may be to increase pollinator attraction. Spittlebug infestation modified interactions with other antagonists in a sex-dependent manner: spittlebugs reduced attack by bud-clipping weevils in hermaphrodites but increased infection by leaf fungi in females. The complex interactions between plant sex, antagonists, and pollinator attraction documented here emphasize the importance of considering sex-differential multi-species interactions in plant sexual evolution. PMID- 21646089 TI - Variation in floral longevity between populations of Campanula rotundifolia (Campanulaceae) in response to fitness accrual rate manipulation. AB - Floral longevity, the time between corolla expansion and senescence, contributes directly and indirectly to a plant's overall fitness. Though mating opportunities for insect-pollinated species often differ among populations, few empirical studies have addressed whether floral longevity varies in a manner consistent with these differences. I conducted experiments at thermally distinct sites to examine whether the prevailing floral longevity model predicted such variation between a montane and an alpine population of Campanula rotundifolia. Staminate phase duration was significantly shorter for montane vs. alpine C. rotundifolia flowers in the presence of pollinators, but significantly longer when pollinators were excluded. Montane flowers had a significantly higher female fitness accrual rate, significantly shorter total longevities, and, unlike alpine flowers, were not pollen-limited. Delaying pollinator access to pistillate phase flowers significantly increased total longevity in alpine flowers only. Significant differences in total longevity between populations resulted from an extended pistillate phase in alpine flowers. Overall, the prevailing model accurately predicted the total floral longevity trends found here. However, I provide novel evidence for geographic and gender-specific plasticity in the floral longevity response to fitness accrual rate variation, suggesting C. rotundifolia populations in this study may be attuned to local schedules of pollinator activity. PMID- 21646090 TI - Phylogenetic analysis of Pinguicula (Lentibulariaceae): chloroplast DNA sequences and morphology support several geographically distinct radiations. AB - The genus Pinguicula is one of the three genera of the carnivorous Lentibulariaceae, comprising approximately 80 species. Phylogeny inference using nucleotide sequences of the chloroplast gene matK and the trnK group II intron, as well as a set of 32 morphological characters revealed five well-supported, major lineages within the genus. These lineages largely reflect radiations in clearly defined geographic regions, whereas most previously recognized sections of the genus are shown to be para- or polyphyletic. A species-rich Mexican Central American-Caribbean clade has the Eurasian P. alpina and an East Asian clade as successive sisters. All three are characterized by a production of flower buds on winter-resting plants, a specific corolla hair structure and a very large corolla lower central lobe. Another diverse clade is composed of species with primarily European distribution including the widespread type species P. vulgaris. For this clade, vegetative reproduction during dormancy is synapomorphic. Species native to SE North America and the South American Andes and a group of Mediterranean and NE Atlantic coast species together appear in a fifth well-supported clade, that is characterized by a tropical-type growth habit. It is the only clade that has reached temperate zones of the southern hemisphere. PMID- 21646091 TI - Dating phylogenetically basal eudicots using rbcL sequences and multiple fossil reference points. AB - A molecular dating of the phylogenetically basal eudicots (Ranunculales, Proteales, Sabiales, Buxales and Trochodendrales sensu Angiosperm Phylogeny Group II) has been performed using several fossils as minimum age constraints. All rbcL sequences available in GenBank were sampled for the taxa in focus. Dating was performed using penalized likelihood, and results were compared with nonparametric rate smoothing. Fourteen eudicot fossils, all with a Cretaceous record, were included in this study for age constraints. Nine of these are assigned to basal eudicots and the remaining five taxa represent core eudicots. Our study shows that the choice of methods and fossil constraints has a great impact on the age estimates, and that removing one single fossil change the results in the magnitude of tens of million years. The use of several fossil constraints increase the probability of approaching the true ages. Our results suggest a rapid diversification during the late Early Cretaceous, with all the lineages of basal eudicots emerging during the latest part of the Early Cretaceous. The age of Ranunculales was estimated to 120 my, Proteales to 119 my, Sabiales to 118 my, Buxales to 117 my, and Trochodendrales to 116 my. PMID- 21646092 TI - Phylogenetic relationships, chromosome and breeding system evolution in Turnera (Turneraceae): inferences from its sequence data. AB - Turnera provides a useful system for exploring two significant evolutionary phenomena-shifts in breeding system (distyly vs. homostyly) and the evolution of polyploids. To explore these, the first molecular phylogeny of Turnera was constructed using sequences of the internal transcribed spacer region (ITS) of nuclear ribosomal DNA for 37 taxa. We attempted to resolve the origins of allopolyploid species using single-strand conformation polymorphism and sequencing of homeologous copies of ITS. Two allohexaploid species possessed putative ITS homeologues (T. velutina and T. orientalis). A phylogenetic analysis to identify progenitors contributing to the origins of these polyploids was unsuccessful, possibly as a result of concerted evolution of ITS. Breeding system evolution was mapped onto the phylogeny assuming distyly to be ancestral in Turnera. Self-compatible homostyly appears to have arisen independently at least three times in Turnera; however, we were not able to determine whether there have been independent origins of homostyly among hexaploid species in series Turnera. Our phylogenetic analyses suggest that series Turnera is monophyletic. Neither series Microphyllae nor Anomalae, however, appear to be monophyletic. Future taxonomic revisions may require new circumscriptions of these latter series. PMID- 21646093 TI - Timing of seed dispersal generates a bimodal seed bank depth distribution. AB - The density of soil seed banks is normally highest at the soil surface and declines monotonically with depth. Sometimes, for a variety of reasons, peak density occurs below the surface but, except in severely disturbed soils, it is generally true that deeper seeds are older. In seasonally dry habitats that develop deep soil cracks during the dry season, it is possible that some seeds fall down cracks and rapidly become deeply buried. We investigated this possibility for three dominant clonal perennials (Scirpus maritimus, S. litoralis, and Juncus subulatus) in the Donana salt marsh, a nontidal marsh with a Mediterranean climate located in southwest Spain. Two species, which shed most of their seed during the dry season and have seeds with low buoyancy, had bimodal viable seed depth distributions, with peak densities at the surface and at 16-20 cm. A third species, which shed most seeds after soil cracks had closed and had seeds with high buoyancy, had viable seeds only in surface soil. Bimodal seed bank depth distributions may be relatively common in seasonally dry habitats with fine-textured soils, but their ecological significance has not been investigated. PMID- 21646094 TI - Development of woody branch attachments in Schefflera (Araliaceae or Apiaceae). AB - Attachment of branches in Schefflera is unusual in that it involves fingerlike woody extensions that originate in the cortex and pass gradually into the woody cylinder of the parent shoot. We tested the hypothesis that these structures could be roots since Schefflera is a hemi-epiphyte with aerial roots. These branch traces originate by secondary development in the many leaf traces (LTs) of the multilacunar node together with associated accessory traces. In the primary condition, the LTs may be described as cortical bundles. Leaves are long persistent and can maintain a primary stem connection across a broad cylinder of secondary xylem. Under the stimulus of branch development, the LTs form a template for secondary vascular development. Because the LT system is broad, with many traces, the branch attachment is also broad. The fingerlike extensions are attached to the surface of the woody cylinder of the parent stem but are progressively obscured as a continuous cambium is formed. Bark tissues are included within the branch axil because of the extended cortical origin of the initial attachment. The results are discussed in the context of branch-trunk unions in tropical plants, an important component of canopy development. PMID- 21646095 TI - Variation in "biodiversity value" of peatmoss species in Sphagnum section Acutifolia (Sphagnaceae). AB - Species are the most common currency by which biodiversity is measured, but species are not equivalent in "biodiversity value" because of differences in phylogenetic history and current population processes. Morphologically defined species in Sphagnum section Acutifolia (Bryophyta) were compared with regard to how phylogenetically distinct each is from its sister species and how much nucleotide variation each encompasses. Comparisons were based on sequence variation at seven nuclear and chloroplast loci. Assignment of collections to morphospecies accounted for about 57-75% of the overall nucleotide variation at the seven loci, but morphospecies differed greatly in how much nucleotide diversity they encompass. In addition, morphospecies varied widely in their genetic distinctiveness, estimated as the length of the stem branch from the most recent common ancestor and numbers of differentially fixed and shared polymorphic nucleotides among taxa. Levels of molecular diversity within morphospecies were not correlated with their degree of isolation. Factors that affected the biodiversity value of species include age, mode of origin, demographic history, and reproductive biology. PMID- 21646096 TI - Differentiation of cellular and biochemical features of the single-cell C4 syndrome during leaf development in Bienertia cycloptera (Chenopodiaceae). AB - The terrestrial plant Bienertia cycloptera has been shown to accomplish C(4) photosynthesis within individual chlorenchyma cells by spatially separating the phases of carbon assimilation into distinct peripheral and central compartments. In this study, anatomical, physiological, and biochemical techniques were used to determine how this unique compartmentation develops. Western blots show ribulose 1,5-bisphosphate carboxylase (Rubisco) (chloroplastic) is present in the youngest leaves and increases during development, while levels of C(4) enzymes-pyruvate,Pi dikinase (chloroplastic), phosphoenolpyruvate carboxylase (PEPC) (cytosol), and NAD-malic enzyme (mitochondrial)-increase later in development. Immunolocalization confirmed this for Rubisco and PEPC. The youngest chlorenchyma cells have a central nucleus surrounded by monomorphic granal chloroplasts containing Rubisco. Later stages show progressive development of a central cytoplasmic compartment enriched with chloroplasts and mitochondria and of a peripheral cytoplasm with chloroplasts. A complex reticulum of connections between the compartments also developed and was characterized. delta(13)C isotope analyses show mature leaves have distinct C(4)-type isotope composition, while the composition in younger leaves is "C(4)-like." Based on the results, this form of single-cell C(4) photosynthesis develops from a common pool of organelles through partitioning to separate compartments, and the development of biochemically and ultrastructurally dimorphic chloroplasts. PMID- 21646097 TI - Sexual reproduction, hybridization, apomixis, and polyploidization in the genus Boechera (Brassicaceae). AB - Of the 340 genera in the Brassicaceae, apomictic reproduction is found only in the North American genus Boechera. We investigated phylogenetic relationships, ability to hybridize, mating system, and ploidy levels of 92 lines sampled from 85 populations and representing 19 Boechera species. Phylogenetic analyses based on chloroplast DNA sequences identified three lineages in the genus. Reciprocal crosses of each line were made to a common sexual diploid B. stricta tester. The resulting F(1) progeny were analyzed for the inheritance of polymorphic microsatellite loci, genome size, and seed production. Intraspecific B. stricta crosses confirmed that this species is mostly diploid and sexual. Interspecific crosses revealed many other species were diploid and sexual and could be successfully hybridized with the tester. We also found obligate and facultative apomictic diploid and triploid lines. De novo F(1) polyploids (either triploids or tetraploids) were derived from the union of nonreduced (from an apomictic parent) and reduced (from the tester) gametes. However, seed production of these F(1) plants was generally low, suggesting a failure in the transmission of apomixis. The creation of a wide array of segregating genetic populations will facilitate future research on the evolution and inheritance of quantitative variation in Boechera. PMID- 21646098 TI - Growth irradiance effects on photosynthesis and growth in two co-occurring shade tolerant neotropical perennials of contrasting photosynthetic pathways. AB - Dieffenbachia longispatha (C3) and Aechmea magdalenae (Crassulacean acid metabolism, CAM) are syntopic, neotropical forest perennials in central Panama that are restricted to shaded habitats. This is of particular interest for A. magdalenae because, like other understory CAM bromeliad species, it appears functionally and structurally to be better suited to life in full sun. Growth irradiance (GI) effects on photosynthesis and growth in both species were explored in the context of sun/shade trade-off concepts largely derived from studies of C3 plants. Potted plants were grown outdoors in 1, 55, and 100% full sun for 5 mo under well-watered conditions. While both species grew faster in high compared to low light, maximum relative growth rates (RGR) in full sun were still extremely slow with A. magdalenae showing a RGR approximately half that of D. longispatha. Photosynthetic capacity increased with GI in D. longispatha but not in A. magdalenae. Aechmea magdalenae responded to GI with shifts in the activity of the different CAM phases. Both species were photoinhibited in full sun, but more so in A. magdalenae. Despite possessing many traits considered adaptive in high light, these results suggest that A. magdalenae is unlikely to attain sufficient growth rates to thrive in productive, high-light habitats. PMID- 21646099 TI - The importance of frictional interactions in maintaining the stability of the twining habit. AB - The stability of twining vines under gravitational loads suggests an important role for friction. The coefficient of friction, MU, between vine stems and wood is high, often five times greater than between leather and wood, as determined by slip tests on an inclined plane. Stem trichomes function like ratchets to facilitate climbing upward (or to facilitate slipping if the stem is inverted). A mathematical model predicts large masses (kg) must be applied to the base of a twining vine to cause slipping. Vines slip as predicted when MU is low and arc length on the pole is short, and they break before slipping when MU is large or arc length is long. In contrast, twining vines are unstable in compression, collapsing when small masses (<10 g) are hung from the top of the vine. However, if the loads are applied below the uppermost gyre, the stabilizing tensional effect dominates. Therefore, in nature vines twining on a cylindrical support are stable under gravitational loads, unless these loads occur near the apex. A corollary is that a short apical coil can hold up large masses of maturing shoot. PMID- 21646100 TI - Ecological factors influencing tetraploid establishment in snow buttercups (Ranunculus adoneus , Ranunculaceae): minority cytotype exclusion and barriers to triploid formation. AB - Polyploid speciation is an ongoing, important source of angiosperm diversity. However, the barriers to polyploid speciation and mechanisms of establishment remain poorly understood for all but a few species. Several factors likely to have influenced tetraploid establishment, including barriers to triploid formation, consequences of mixed-ploidy pollen loads, and the reproductive success of the minority cytotype, were examined in snow buttercups (Ranunculus adoneus) through a series of pollination and transplant experiments. Tetraploid snow buttercups do not have stigmatic barriers to pollen from diploid plants, nor does pollen from tetraploid plants have an advantage over pollen from diploids when on tetraploid stigmas. Tetraploid plants transplanted into a diploid population produced 50% fewer seeds than tetraploid plants in a tetraploid population. Intrinsic barriers to triploid formation were relatively weak, but few triploid seeds formed when mixed-ploidy pollen was present. Fecundity of triploid plants was very low, and no tetraploid offspring resulted. These results indicate that in snow buttercups, a triploid plant will contribute 0.8% of the tetraploid seeds of a minority tetraploid plant making it a minor contributor to the demographics of tetraploid establishment. The reproductive costs facing minority cytotype plants may explain the previously observed spatial segregation in snow buttercups. PMID- 21646101 TI - Reproductive developmental complexity in the African oil palm (Elaeis guineensis, Arecaceae). AB - Species of the palm family (Arecaceae) are remarkably diverse in their inflorescence and floral morphologies, which make them a particularly interesting group for studies of reproductive development and its evolution. Using light and scanning electron microscopy, we describe inflorescence and flower development in the African oil palm Elaeis guineensis from the initiation of the inflorescence meristem to flower maturity. In mature palms, the inflorescence develops over 2-3 years and is characterized by individual stages within which differentiation may be either relatively slow, as in the case of early inflorescence meristem development, or rapid, as in the case of flower organogenesis. The female inflorescence bears floral triads composed of single pistillate flowers flanked by two abortive staminate flowers, whereas the male inflorescence contains single functional staminate flowers. This suggests a possible evolutionary movement from an ancestral hermaphrodite inflorescence form containing fully functional floral triads to the situation of temporal dioecy observed at present. Wild type flowers are compared to those bearing an epigenetic homeotic abnormality, known as mantled, involving an alteration of the identity of the organs in the fertile and sterile androecium. PMID- 21646102 TI - Inbreeding depression in perennial Lychnis viscaria (Caryophyllaceae): effects of population mating history and nutrient availability. AB - We studied inbreeding depression in a perennial plant, Lychnis viscaria, in three populations differing in their inbreeding history and population size by measuring several traits at two nutrient levels over the plant's life cycle. The observed levels of inbreeding depression (cumulative inbreeding depression, from 0.057 to 0.629) were high for a plant with a mixed mating system. As expected, the population with a low level of isozyme variation expressed the least inbreeding depression for seed germination. Highest inbreeding depression for germination was found in the largest and genetically most variable population. No clear differences between populations in expression of inbreeding depression in the later life stages were found. The population level inbreeding depression varied with the nutrient conditions and among populations and life stages, but we found no evidence that inbreeding depression increased with lower nutrient availability. These results emphasize the importance of measuring inbreeding depression under several environmental conditions and over life stages. PMID- 21646103 TI - Functional associations of floret and inflorescence traits among grass species. AB - The aerodynamics of wind pollination selects for an intimate relation between form and function in anemophilous plants. Inflorescence architecture and floral morphology vary extensively within the Poaceae, but the functional implication of this variation remains largely unknown. Here we quantify associations between floret, culm, and inflorescence characteristics for 25 grass species in Kananaskis, Alberta, Canada, and consider whether different architectures may implement unique mechanisms to aid pollination. The species cluster into four categories defined by all combinations of floret size (small vs. large) and inflorescence architecture (compact vs. diffuse). Species differed significantly for all 18 traits that we measured, with 12 traits differing only between floret size classes, three differing only between inflorescence types, and three differing among both (independently or by an interaction). Based on these morphological associations, we discuss the aerodynamic and functional consequences of each category for wind pollination. The independence of inflorescence and floral traits has probably allowed exploration of all possible combinations of inflorescence architecture and floret size during the evolution of the Poaceae. PMID- 21646104 TI - Inbreeding depression and selfing rate of Ipomoea hederacea var. integriuscula (Convolvulaceae). AB - Inbreeding depression and selfing rate were investigated in the self-compatible vine Ipomoea hederacea to assess the variability of the breeding system. Inbreeding depression differed between populations and the magnitude varied at germination, growth (as measured by aboveground biomass), and reproductive potential. Plants from Macon County, Alabama, USA, had significant inbreeding depression (31%) at germination, but no significant inbreeding depression for aboveground biomass or number of reproductive structures (buds and flowers) at 45 d post germination in the greenhouse or in the field. Plants from Morgan County, Alabama, however, had significant inbreeding depression (>50%) for all three stages in the greenhouse. In allozyme comparisons, five of the 11 I. hederacea populations surveyed had high selfing rates (66.66-92.53%) and high levels of homozygosity (F(IS) = 0.500-0.861) in 2003, and three of four populations surveyed in 2004 had selfing rates that exceeded 50%. High selfing rates, high levels of homozygosity, and low levels of inbreeding depression suggest that inbreeding depression may not present a significant barrier to the transmission of selfing alleles in some populations of I. hederacea, but does not account for the maintenance of a mixed mating system in other populations. PMID- 21646105 TI - Inversostyly: a new stylar polymorphism in an oil-secreting plant, Hemimeris racemosa (Scrophulariaceae). AB - A new kind of stylar polymorphism, provisionally called inversostyly, is described. The polymorphism occurs in Hemimeris racemosa (Scrophulariaceae), a common annual herb of the Cape region of South Africa. Most populations are dimorphic for style orientation: the style alternates with the two stamens and is deflected either upwards or downwards. Thus, there is reciprocal placement of the style and stamens in a vertical plane in zygomorphic flowers. The flowers are symmetrical, and the floral parts do not vary in length. All flowers on a given plant are of the same stylar orientation. Pollination is by specialized oil collecting bees (Rediviva spp.), which carry the pollen of the two morphs separately in discrete anterior or posterior locations on the underside of the body. Most inversostylous populations have a slightly higher proportion of the style-down morph, and this bias increases with decreasing pollinator abundance. In contrast with inversostylous populations, all individuals in homostylous populations of H. racemosa have the style and the stamens clustered together in the down position and high levels of autogamous seed set. Homostylous populations of H. racemosa, as well as the homostylous species Hemimeris sabulosa, occur where oil-collecting bees are less abundant. PMID- 21646106 TI - Intra- and interspecific phylogenetic relationships among diploid Triticum Aegilops species (Poaceae) based on base-pair substitutions, indels, and microsatellites in chloroplast noncoding sequences. AB - This study analyzes intra- and interspecific variation in chloroplast DNA (cpDNA) in diploid Triticum-Aegilops species. This analysis focused on DNA sequence variation in noncoding regions of cpDNA, which included base-pair substitutions, insertion/deletions (indels, 50 loci pooled), microsatellites (7 loci pooled), and inversions. Nine of 13 Triticum-Aegilops species were successfully identified and genotyped using these data. Sixty-two haplotypes were detected in 115 accessions of 13 diploid species. Because of the large number of characters examined, novel deep relationships within and among Triticum-Aegilops species could be identified and evaluated. Phylogenetic trees for the genus Triticum Aegilops were constructed with Hordeum vulgare and Dasypyrum villosum as outgroups, and the results were compared to previous studies. These data support the following inferences: (1) Aegilops species should be included in Triticum; (2) groups D, T, M, N, U, and section Sitopsis (except Ae. speltoides) underwent speciation concurrently, but most diploid species evolved independently; (3) Ae. mutica does not occupy a basal position in Triticum-Aegilops; (4) Ae. speltoides is in a basal position and differs significantly from other Sitopsis species; (5) Ae. caudata is polyphyletic in all trees; (6) the genus Aegilops is paraphyletic with Secale. PMID- 21646107 TI - Rapid speciation and the evolution of hummingbird pollination in neotropical Costus subgenus Costus (Costaceae): evidence from nrDNA ITS and ETS sequences. AB - We estimate phylogenetic relationships and the biogeographic and pollination history of Costus subgenus Costus (Costaceae) using sequence data from the internal and external transcribed spacer (ITS and ETS) regions of 18S-26S nuclear ribosomal DNA. The African members of the subgenus form a series of lineages basal to a monophyletic neotropical species radiation. The neotropical species have large, showy flowers visited by either euglossine bees or hummingbirds. The hummingbird pollination syndrome is supported as a derived character state from the bee pollination syndrome, and we estimate that it has evolved independently seven or more times in the neotropics. A molecular clock approach suggests that diversification of the neotropical clade has been recent and rapid and that it coincides with dramatic climatic and geologic changes, Andean orogeny, and the closing of the Panama isthmus that occurred in the Pliocene and Pleistocene epochs. We propose a scenario for the diversification of Costus, in which rapid floral adaptation in geographic isolation and range shifts in response to environmental changes contribute to reproductive isolation among close relatives. We suggest that these processes may be common in other recently diversified plant lineages centered in Central America or the Northern Andean phytogeographic region. PMID- 21646108 TI - Phylogeny of Flaveria (Asteraceae) and inference of C4 photosynthesis evolution. AB - A well-resolved phylogeny of Flaveria is used to infer evolutionary relationships among species, biogeographical distributions, and C(4) photosynthetic evolution. Data on morphology, life history, and DNA sequences (chloroplastic trnL-F, nuclear ITS and ETS) for 21 of 23 known species were collected. Each data set was analyzed separately and in combination using maximum parsimony and Bayesian analyses. The phylogeny of Flaveria is based on the combined analysis of all data. Our phylogenetic evidence indicates that C(3) Flaveria are all basal to intermediate (C(3)-C(4) and C(4)-like) and fully expressed C(4) Flaveria species. Two strongly supported clades (A and B) are present. Using this phylogeny, we evaluate the current systematics of the genus and suggest the removal and reevaluation of certain taxa. We also infer the center of origin and dispersal of Flaveria species. Multiple origins of photosynthetic pathway intermediacy in Flaveria are recognized. C(3)-C(4) intermediacy has evolved twice in the genus and is found to be evolutionarily intermediate in clade A, but not necessarily in clade B. C(4)-like photosynthesis is also derived once in each clade. In addition, fully expressed C(4) photosynthesis may have evolved up to three times within clade A. PMID- 21646109 TI - Problems in measuring among-family variation in inbreeding depression. AB - Understanding the sources of variation in inbreeding depression within populations is important for understanding the evolution of selfing rates. At the population level, inbreeding depression is due to decreased heterozygosity caused by inbreeding, which decreases overdominance and increases the frequency of expression of recessive deleterious alleles. However, within individual families inbreeding has two distinct consequences: it reduces heterozygosity and it restricts the alleles present in offspring to those present in the parent. Outcrossing both increases heterozygosity and brings new alleles into a family (compared to the alleles present if the plant is self-pollinated). Both consequences of inbreeding affect offspring fitness, but the most common experimental design used to measure among-family variation in inbreeding depression cannot distinguish them. The result is that variance in inbreeding depression among families is confounded by genetic variation in the traits being measured. Also, correlations (among families) between measures of inbreeding depression or between inbreeding depression and mean trait values are confounded by genetic variation in the traits being measured. I conclude that more complex crossing designs that allow estimation of breeding values for individual families are required to accurately detect and measure among-family variation in inbreeding depression. PMID- 21646110 TI - Development of the calcium oxalate crystal macropattern in pomegranate (Punica granatum, Punicaceae). AB - Oxalate crystals are very common in angiosperms, but few descriptions of their macropattern (crystal types, their tissue distribution, and development) exist. Because unusually large prismatic crystals and druses, are known from pomegranate, we traced the development of crystal macropattern in various-aged leaf samples from a living plant and from herbarium specimens using unstained whole mounts (some bleached and cleared), stained leaf samples, and leaf and stem cross sections. Preparations were viewed with bright-field light microscopy and with crossed polarizers. Prismatics appear first in the subapical mid-mesophyll layer of a leaf 650 MUm long. Additional prismatics form basipetally in the enlarging lamina. A preemptive wave of small prismatics appears basipetally in the midrib. Druses form secondarily acropetally in petiole and midrib, while existing lamina prismatics enlarge and new ones develop among them in mid mesophyll. Prismatics produced early expand vertically, and many eventually extend from epidermis to epidermis. Later-formed prismatics attain intermediate sizes. No crystals form along lamina veins, but in older leaves, druses occur in spongy mesophyll, mostly near major vein junctions. In the stem, druses are restricted to phloem fibers. No phloem fibers occur in the leaf trace or midvein; therefore, petiolar and midrib druses are only in parenchyma, not in phloem. PMID- 21646111 TI - DNA content and expression of genes related to cell cycling in developing Gossypium hirsutum (Malvaceae) fibers. AB - The cell cycle in cotton (Gossypium hirsutum) fibers is poorly understood. The objective of this study was to evaluate the cell cycle status and DNA content in developing cotton fibers. The DNA content and cell cycle distribution in fiber and hypocotyl cells were determined by flow cytometry. Expression levels of minichrosomal maintenance protein (mcm), cyclin B, and a retinoblastoma-like protein (rb) genes were determined with real-time PCR in fibers and dividing and nondividing tissues. No endoreduplication occurred, nor did genome size or percentage of G1-phase nuclei differ between hypocotyls and fibers. Approximately 13 and 17% of fiber nuclei were in the S phase in 14 days after anthesis (d) fibers and 25 d fibers, respectively. The mcm and cyclin B were expressed at higher levels in fibers than in mature leaves, but expression levels in fibers were less than 15% of meristematic tissues. Rb was expressed in fibers at levels less than 50% of mature leaves or meristematic tissues. Based on an apparent increase in S-phase cells as fibers mature and the low level of expression of genes associated with cell cycle progression, we conclude that S-phase arrest occurs in developing cotton fiber. PMID- 21646112 TI - Vegetation patterns 25 years after the eruption of Mount St. Helens, Washington, USA. AB - In 2004, we surveyed the vegetation on Mount St. Helens to document changes since 1992. We asked how communities differentiate and if they develop predictable relationships with local environments. We sought evidence from links between species and environment and changes in community structure in 271 250-m(2) plots. The habitats of the seven community types (CTs) overlapped broadly. Ordination methods demonstrated weak correlations among species distributions and location, elevation, and surface variables. Comparisons to 1992 by habitat demonstrated a large increase in plant cover and substantial development of vegetation structure. Pioneer species declined while mosses increased proportionately leading to more pronounced dominance hierarchies in most habitats. In Lupinus colonies, dominance declined, and diversity increased due to the increased abundance of formerly rare species. On once barren sites, dominance increased, but diversity changed slightly, which suggested the incipient development of competitive hierarchies. Weak correlations between vegetation and the environment suggested that initially stochastic establishment patterns had not yet been erased by deterministic factors. A vegetation mosaic that is loosely controlled by environmental factors may produce different successional trajectories that lead to alternative stable communities in similar habitats. This result has implications for restoration planning. PMID- 21646113 TI - Erratum. PMID- 21646114 TI - A new Late Cretaceous ginkgoalean reproductive structure Nehvizdyella gen. nov. from the Czech Republic and its whole-plant reconstruction. AB - During the Mesozoic Era, gingkoaleans comprised a diverse and widespread group. Here we describe ginkgoalean fossils in their facies context from the Late Cretaceous (Cenomanian) Peruc-Korycany Formation of the Czech Republic and present a reconstruction of tree architecture and ecology. Newly described in this study is the ovuliferous reproductive structure, Nehvizdyella bipartita gen. et sp. nov. (Ginkgoales). This ovuliferous organ consists of a bifurcating axis, terminated by large cupule-like structures, probably homologous to the collar of the recent Ginkgo. Each cupule encloses an orthotropous ovule. In specimens with the early developmental stages preserved, the entire ovule and young seed, with the exception of the micropylar area, is embedded in the cupule. Mature seeds consist of sclerotesta and sarcotesta. Monosulcate pollen grains of Cycadopites type are found adhering to the seeds. Although similar to Ginkgo in terms of its large size and reduced number of seeds, N. bipartita differs from the extant genus in having ovules completely enclosed in a cupule-like structure. The co occurrence of N. bipartita with ginkgoalean leaves of Eretmophyllum obtusum (Velenovsky) Kvacek, J., ginkgoalean short shoots of Pecinovicladus kvacekii Falcon-Lang, and ginkgoalean trunk wood of Ginkgoxylon gruettii Pons and Vozenin Serra in monodominant taphocoenoses at four geographically distant localities suggests that these remains all belong to one plant. This is supported by the close morphological and anatomical similarity between the different organs. Facies analysis of plant assemblages indicates that our Cretaceous tree occupied a water-stressed coastal salt marsh environment. It therefore represents the first unequivocal halophyte among the Ginkgoales. PMID- 21646115 TI - Embolized conduits of rice (Oryza sativa, Poaceae) refill despite negative xylem pressure. AB - Embolism reversal in rice plants was studied by testing the plant's ability to refill embolized conduits while xylem pressures were substantially negative. Intact, potted plants were water-stressed to a xylem pressure of -1.88 +/- 0.1 MPa and a 66.3 +/- 3.8% loss of xylem conductivity (PLC) by cavitation. Stressed plants were carefully rewatered, allowing xylem pressure to rise, but not above the theoretical threshold of c. -0.15 MPa for embolism collapse. Despite xylem pressures being more negative than this threshold, the PLC fell significantly (28.5 +/- 5.6%), indicating the refilling of vessels. Above c. -1.0 MPa, almost all plants regained their maximum hydraulic conductivity. Dye uptake experiments showed the same pattern of embolism refilling despite negative pressure. Refilling was prevented in plants that were light-starved for 5 d, suggesting the unknown mechanism is dependent on metabolic energy. Results are among the first showing that herbaceous plants can reverse embolism without bulk xylem pressures rising near or above atmospheric. PMID- 21646116 TI - Genic diversity, genetic structure, and mating system of Brewer spruce (Pinaceae), a relict of the Arcto-Tertiary forest. AB - Brewer spruce (Picea breweriana), a relict of the widespread Arcto-Tertiary forests, is now restricted to a highly fragmented range in the Klamath Region of California and Oregon. Expected heterozygosity for 26 isozyme loci, averaged over 10 populations, was 0.121. More notable than the relatively high level of diversity when compared to other woody endemics was the strong decrease in expected heterozygosity with latitude. Differentiation (F(ST)) was 0.152, higher than values for many north temperate conifers with larger distributions. The number of migrants per generation (Nm) was 1.34 or 2.70, depending on the method of estimation. Inbreeding appeared low; F(IS) was only 0.003, in agreement with multilocus population outcrossing rates (t(m)), which were generally well above 0.90. No difference in t(m) was found between isolated vs. clustered trees. However, the number of seeds per cone was greatest in the densest populations; t(m) is a measure of effective outcrossing after mortality in the embryonic stage, whereas a reduced number of seeds per cone indicates self pollination. Selfing increased after logging; outcrossing rate before logging was 0.961 and after logging, 0.756. Despite Brewer spruce's narrow, fragmented distribution, the outlook for its conservation was good, with the exception of possible negative effects of logging. PMID- 21646117 TI - Determinants of species rarity: population growth rates of species sharing the same habitat. AB - Determining differences between common and rare species is commonly used to identify factors responsible for rarity. Existing studies, however, suffer from two important drawbacks. First, studies compare species that are closely related phylogenetically but occupy different habitats. Second, these studies concentrate on single life history traits, with unknown relevance for population growth rates. Complete life cycles of one rare and one common Cirsium species sharing the same habitat were compared. Population growth rate was slightly lower in the rare species, translating into a large difference in local extinction probability. Seed predation intensity did not differ between species. However, it can be demonstrated that in connection with the data on complete demography, seed predation is the key factor causing a lower population growth rate in the rare species. These results are the first estimation of factors responsible for commonness or rarity of plants in terms of population growth rate without confounding differences in ecology. They demonstrate that conclusions based on single traits may be misleading and that only a comparison based on a complete life cycle can provide unequivocal evidence for concluding which factors are really those responsible for species commonness or rarity. PMID- 21646118 TI - The role of pollination level on the reproduction of females and hermaphrodites in the gynodioecious plant Gypsophila repens (Caryophyllaceae). AB - In gynodioecious plant species, females are expected to have more resources available for maturing seeds because pistillate flowers are smaller, do not produce pollen, and are thus less costly that perfect flowers. The potential female advantage arising from more abundant resources is, however, likely to vary depending on whether seed production is limited by resource or pollen availability. Here we experimentally investigated the influence of pollen and resource limitation on female advantage in a gynodioecious species using two levels of pollination. Total seed production of females was always greater than that of hermaphrodites: females produced more flowers and more fruits that contained similar numbers of seeds of similar mass. Under low pollination, female and hermaphrodite plants allocated resources to increased flower production rather than to increased seed size or quality. We did not detect any influence of pollen or resource limitation on female advantage, which remained similar under low (= abundant resources) and full pollination. Outcrossed fruits performed better than selfed fruits when the same plant received both selfed and outcrossed pollen on different flowers. These differences were not greater under high pollination, possibly because resources available for each fruit did not differ between our pollen intensity treatments. PMID- 21646119 TI - Reproductive investment in male and female Eurya japonica (Theaceae) at tree and branch levels. AB - Intersexual differences in reproductive investments (RIs) (dry mass of reproductive tissue) at tree and branch levels in Eurya japonica were examined during two successive years. Mean total RIs per tree for males and females (adjusted for the mean trunk diameter in the combined sample trees, 25.7 mm) were 3.47 and 5.67 g dry mass. Females generally allocated about 1.6 times more biomass per tree to reproduction than males. On the other hand, male total RI per terminal branch averaged 51.6 mg dry mass and female averaged 226.6 mg. Females generally allocated about 4.4 times more biomass per terminal branch to reproduction than males. Thus, the magnitude of sexual difference at the tree level was much lower than that at the branch level. There were negative correlations between interyear fluctuation of total RI and stem diameter for both sexes. Interyear fluctuation of RI was greater for females than males in all size categories. This study revealed that conclusions from tree measures of RI differed from branch measures and suggested the importance of evaluating the average RI at the tree level for woody plants. I discussed the importance in adopting an effective sampling strategy for evaluating the RI at the tree level. PMID- 21646120 TI - Chloroplast DNA phylogeny and phylogeography of the North American plums (Prunus subgenus Prunus section Prunocerasus, Rosaceae). AB - The North American plums are a closely related group that are not easily circumscribed, have overlapping morphologies, and are known to hybridize. We previously showed that the North American plums are a closely related, monophyletic group of taxa with little to no cpDNA sequence divergence between taxa. In that study, we came to the unanticipated conclusion that relationships inferred among the taxa contrast sharply with previously defined groups based on morphological characters. Here the aim was to determine if the primary cpDNA haplotypes identified in our earlier study are confined to the taxa in which they were initially observed. The cpDNA rpL16 intron was sequenced for 207 accessions of the 17 North American plum taxa plus Prunus texana. The results show that many taxa contain more than one of the three primary cpDNA haplotypes. Aside from the results found in sect. Prunocerasus, this study has broader implications for phylogenetics in general. The common practice of choosing a single exemplar to represent a taxon can be profoundly misleading in closely related groups. In hindsight, the possibility existed in our earlier study that we could have chosen a different combination of exemplars, which could have resulted in a different inferred phylogeny. PMID- 21646121 TI - Genetic variation and relationships among Ulex (Fabaceae) species in southern Spain and northern Morocco assessed by chloroplast microsatellite (cpSSR) markers. AB - Genetic variation in 27 populations of Ulex species from southern Spain and northern Morocco (Betic-Rif arc) was assessed using 11 chloroplast microsatellite (cpSSR) markers, which revealed 47 different haplotypes. These nonrecombinant, haploid markers allow measurement of genetic variation in closely related species of Ulex where molecular phylogenetic analyses have not provided a clear view of interspecific relationships. Discriminant analysis indicates that the haplotypes are useful to differentiate among species, and analysis of molecular variance (AMOVA) shows high levels of differentiation among populations. The minimum spanning tree (MST), that represents the connections between the haplotypes, suggests that the eastern Rifean U. africanus haplotypes are more genetically related than those from southern Spain. The latter may have lost genetic diversity while colonizing new habitats, eventually differentiating into U. baeticus and U. scaber. Hybridization between these populations, followed by polyploidization, may have originated the tetraploids (U. congestus and U. borgiae) that colonized new habitats associated with acidic rocks. Separate groupings of U. scaber populations may indicate multiple origins from different stocks. Diversification in this group of Ulex species could be related to the opening of the Alboran Sea by Middle Miocene, when the populations from Morocco and Spain became isolated from each other. PMID- 21646122 TI - Relationships within tribe Lycieae (Solanaceae): paraphyly of Lycium and multiple origins of gender dimorphism. AB - We infer phylogenetic relationships among Lycium, Grabowskia, and the monotypic Phrodus microphyllus, using DNA sequence data from the nuclear granule-bound starch synthase gene (GBSSI, waxy) and the chloroplast region trnT-trnF. This is the first comprehensive molecular phylogenetic study of tribe Lycieae (Solanaceae). In addition to providing an understanding of evolutionary relationships, we use the phylogenetic hypotheses to frame our studies of breeding system transitions, floral and fruit evolution, and biogeographical patterns within Lycieae. Whereas Lycium is distributed worldwide, Phrodus and the majority of Grabowskia species are restricted to South America. Tribe Lycieae is strongly supported as monophyletic, but Lycium likely includes both Grabowskia and Phrodus. Results also suggest a single dispersal event from the Americas to the Old World, and frequent dispersal between North and South America. The diversity of fruit types in Lycieae is discussed in light of dispersal patterns and recent work on fruit evolution across Solanaceae. Dimorphic gender expression has been studied previously within Lycium, and results indicate that transitions in sexual expression are convergent, occurring multiple times in North America (a revised estimate from previous studies) and southern Africa. PMID- 21646123 TI - Molecular systematics of the trans-Pacific alpine genus Oreomyrrhis (Apiaceae): phylogenetic affinities and biogeographic implications. AB - The alpine ecosystem is the only terrestrial biogeographic unit that is distributed globally. Studying phylogenetics of the plant species in this widespread ecosystem can provide insights into the historical biogeographic processes that have shaped the global biodiversity. The trans-Pacific disjunct alpine genus Oreomyrrhis (Apiaceae) was investigated using nrDNA ITS sequences to test the taxonomic and biogeographic hypotheses. Phylogenetic analyses using maximum parsimony, maximum likelihood, and Bayesian inference revealed that species of Oreomyrrhis form a weakly supported monophyletic clade that is nested within Chaerophyllum sect. Chaerophyllum (subtribe Scandicinae, tribe Scandiceae). The optimal solutions of dispersal-vicariance analysis indicate that the ancestor of Chaerophyllum sect. Chaerophyllum (including Oreomyrrhis) was distributed in Eurasia and subsequently dispersed to North America and southern Pacific Rim. Based on dating using ITS sequence variation, these dispersal events were most likely recent, probably during late Tertiary to Quaternary. The structure of the ITS haplotype network suggests that a rapid range expansion via long-distance dispersal had been crucial in generating the trans-Pacific disjunction of Oreomyrrhis. Furthermore, evolution toward smaller mericarp size and a transition from outcrossing to selfing during Oreomyrrhis's evolution might have increased the chances for long-distance dispersal, facilitating its range expansion and occupation on alpine environments. PMID- 21646124 TI - Chloroplast DNA phylogeny of the woody Sonchus alliance (Asteraceae: Sonchinae) in the Macaronesian Islands. AB - Determining accurate phylogenetic relationships among the members of the woody Sonchus alliance presents challenges because of an insufficient level of molecular variation and the convergent evolution of similar morphological traits in island settings. To obtain a better resolved phylogeny and to test the potential role of hybridization and introgression, we sequenced all members of the alliance with multiple populations for the ITS of nrDNA and over 4000 base pairs of coding and noncoding regions of cpDNA. The cpDNA phylogeny is not well resolved in the core members of the alliance (i.e., subg. Dendrosonchus and genus Taeckholmia), but like the ITS tree, it has identified basal lineages of monotypic genera. The cpDNA data set was not significantly different from that of ITS, and subsequent combined analysis provided a better resolved and supported phylogeny within the alliance. The combined ML tree identified the same basal lineages, suggested nonmonophyly of Dendrosonchus and Taeckholmia, and did not support either Boulos' or Aldridge's infrasubgeneric classification system. Assessment of the role of hybridization and introgression was limited due to poor resolution in the cpDNA phylogeny. The combined analysis supports a Gran Canaria origin for the alliance and two subsequent long distance dispersal events to Madeira and Cape Verde islands. PMID- 21646125 TI - Evolutionary relationships among Pinus (Pinaceae) subsections inferred from multiple low-copy nuclear loci. AB - Sequence data from nrITS and cpDNA have failed to fully resolve phylogenetic relationships among Pinus species. Four low-copy nuclear genes, developed from the screening of 73 mapped conifer anchor loci, were sequenced from 12 species representing all subsections. Individual loci do not uniformly support either the nrITS or cpDNA hypotheses and in some cases produce unique topologies. Combined analysis of low-copy nuclear loci produces a well-supported subsectional topology of two subgenera, each divided into two sections, congruent with prior hypotheses of deep divergence in Pinus. The placements of P. nelsonii, P. krempfii, and P. contorta have been of continued systematic interest. Results strongly support the placement of P. nelsonii as sister to the remaining members of sect. Parrya, suggest a moderately well-supported and consistent position of P. krempfii as sister to the remaining members of sect. Quinquefoliae, and are ambiguous about the placement of P. contorta. A successful phylogenetic strategy in Pinus will require many low-copy nuclear loci that include a high proportion of silent sites and derive from independent linkage groups. The locus screening and evaluation strategy presented here can be broadly applied to facilitate the development of phylogenetic markers from the increasing number of available genomic resources. PMID- 21646126 TI - The compensatory responses of an understory herb to experimental damage are habitat-dependent. AB - Canopy gap formation strongly influences the diversity and dynamics of both tropical and temperate forests. It is often viewed as inherently beneficial for understory plants, primarily because growth and flowering are enhanced when light is no longer a limiting resource. It can also be detrimental, however, because plants can be damaged by falling crowns or branches. To elucidate the responses of the Amazonian understory herb Heliconia acuminata to damage sustained during gap formation, we transplanted both experimentally damaged and control plants to canopy gaps and the forest understory. We then measured their patterns of growth and biomass allocation 10 mo later. Despite losing approximately 50% of their leaf area, all damaged plants survived the duration of our experiment. Furthermore, damaged plants transplanted to gaps had relative growth rates that far exceeded those of undamaged plants in both gaps and the forest understory. There were also significant interactions between damage and destination habitat type on root to shoot ratios and leaf-area ratios. Our results suggest the ability of herbaceous plants to recover from damage, as well as patterns of post damage biomass allocation, may be habitat-dependent in ways that have previously remained unexplored. PMID- 21646127 TI - Comparative structure of the labellum in Ophrys fusca and O. lutea (Orchidaceae). AB - The morphology and anatomy of the labellar epidermal cells and the way in which they are arranged are described in an attempt to locate and characterize the osmophore in Ophrys fusca and O. lutea. The micromorphology of the labellum of these two species is similar. Four types of epidermal cells are present on the adaxial surface of the labellum. Long unicellular trichomes with straight tips cover the basal region of the labellum, whereas short unicellular trichomes with polygonal flattened bases form the reflective median speculum. The apical region of the labellum possesses a villous indumentum of long acuminate trichomes with bent or sinuate tips. Large smooth-walled, dome-shaped papillae occur on the margins and on the distal region of the abaxial surface of the labellum. These remarkable papillae have high polarity; the protoplasm at the apex of each cell contains several small vacuoles, while a prominent nucleus surrounded by numerous hypertrophied amyloplasts occurs at the opposite end of the cell. Positive reactions to Vogel's staining test and to Sudan black B enabled us to conclude that the osmophores of both species are composed of these peculiar secretory epidermal cells and by two or three subsecretory layers of parenchyma cells. PMID- 21646128 TI - The evolution of unisexual flowers: morphological and functional convergence results from diverse developmental transitions. AB - Unisexual flower morphology was examined within a phylogenetic context in order to identify developmental transitions associated with the multiple origins of dioecy in flowering plants. Historically, two categories of unisexual flowers have been recognized: type I flowers exhibit rudiments of the nonfunctional organ type, while type II flowers bear no vestigial sexual organs. Mapping of these flower types onto a composite phylogeny shows that type II morphology is homoplasious and has resulted from at least four distinct evolutionary developmental pathways. The historical assignment of unisexual flowers into only two morphological types has masked important developmental and evolutionary dynamics. PMID- 21646129 TI - Inter- and intraspecific structural variations among intervascular pit membranes, as revealed by field-emission scanning electron microscopy. AB - The structure of the intervascular pit membranes of four dicotyledonous species (Salix sachalinensis, Betula platyphylla var. japonica, Acer mono, and Fraxinus mandshurica var. japonica) was examined by field-emission scanning electron microscopy. The intervascular pit membranes of F. mandshurica var. japonica had thin surface layers and a dense middle layer, while no similar middle layer was detectable in the other three species. In F. mandshurica var. japonica, the entire area of each pit membrane was densely covered with microfibrils. In the other three species, by contrast, openings were found in the pit membranes. In some of the intervascular pit membranes of S. sachalinensis, B. platyphylla var. japonica, and A. mono, microfibrils were sparsely interwoven in small areas of the pit membranes and openings of up to several hundred nanometers in diameter were present in such regions. These porous regions tended to be located in peripheral areas of pit membranes. In S. sachalinensis and B. platyphylla var. japonica, ethanol-soluble extracts, whose chemical nature and function remain unknown, were heavily distributed over the intervascular pit membranes. Our observations suggest that the structure of intervascular pit membranes is more complicated than has previously been acknowledged. PMID- 21646130 TI - The cost of carnivory for Darlingtonia californica (Sarraceniaceae): evidence from relationships among leaf traits. AB - Scaling relationships among photosynthetic rate, foliar nutrient concentration, and leaf mass per unit area (LMA) have been observed for a broad range of plants. Leaf traits of the carnivorous pitcher plant Darlingtonia californica, endemic to southern Oregon and northern California, USA, differ substantially from the predictions of these general scaling relationships; net photosynthetic rates of Darlingtonia are much lower than predicted by general scaling relationships given observed foliar nitrogen (N) and phosphorus (P) concentrations and LMA. At five sites in the center of its range, leaf traits of Darlingtonia were strongly correlated with elevation and differed with soil calcium availability and bedrock type. The mean foliar N : P of 25.2 +/- 15.4 of Darlingtonia suggested that these plants were P-limited, although N concentration in the substrate also was extremely low and prey capture was uncommon. Foliar N : P stoichiometry and the observed deviation of Darlingtonia leaf traits from predictions of general scaling relationships permit an initial assessment of the "cost of carnivory" in this species. Carnivory in plants is thought to have evolved in response to N limitation, but for Darlingtonia, carnivory is an evolutionary last resort when both N and P are severely limiting and photosynthesis is greatly reduced. PMID- 21646131 TI - Seed storage conditions change the germination pattern of clonal growth plants in Mediterranean salt marshes. AB - The effect of salinity level and extended exposure to different salinity and flooding conditions on germination patterns of three salt-marsh clonal growth plants (Juncus subulatus, Scirpus litoralis, and S. maritimus) was studied. Seed exposure to extended flooding and saline conditions significantly affected the outcome of the germination process in a different, though predictable, way for each species, after favorable conditions for germination were restored. Tolerance of the germination process was related to the average salinity level measured during the growth/germination season at sites where established individuals of each species dominated the species cover. No relationship was found between salinity tolerance of the germination process and seed response to extended exposure to flooding and salinity conditions. The salinity response was significantly related to the conditions prevailing in the habitats of the respective species during the unfavorable (nongrowth/nongermination) season. Our results indicate that changes in salinity and hydrology while seeds are dormant affect the outcome of the seed-bank response, even when conditions at germination are identical. Because these environmental-history-dependent responses differentially affect seed germination, seedling density, and probably sexual recruitment in the studied and related species, these influences should be considered for wetland restoration and management. PMID- 21646132 TI - Mechanisms for tolerating freeze-thaw stress of two evergreen chaparral species: Rhus ovata and Malosma laurina (Anacardiaceae). AB - The response to freeze-thaw stress was examined for two co-occurring evergreen species, Malosma laurina and Rhus ovata. Laboratory and field experiments on adults and seedlings were made in the spring and winter in 1996 and again on adults in 2003 and 2004. Laboratory and field results indicated that the stem xylem for adults of M. laurina and R. ovata were similarly susceptible to freezing-induced cavitation (percentage loss of conductivity = 92 +/- 2.6% for R. ovata and 90 +/- 4.2% for M. laurina at <= -6 degrees C). In contrast, leaves of M. laurina were more susceptible to freezing injury than leaves of R. ovata. Among seedlings in the field, leaves of M. laurina exhibited freezing injury at 4 degrees C and total shoot mortality at -7.2 degrees C, whereas co-occurring seedlings of R. ovata were uninjured. Surprisingly, R. ovata tolerates high levels of freezing-induced xylem embolism in the field, an apparently rare condition among evergreen plants. Rhus ovata avoids desiccation when xylem embolism is high by exhibiting low minimum leaf conductance compared to M. laurina. These results suggest a link between minimum leaf conductance and stem hydraulics as a mechanism permitting the persistence of an evergreen leaf habit in freezing environments. PMID- 21646133 TI - Pollen movement in a natural population of Arisaema serratum (Araceae), a plant with a pitfall-trap flower pollination system. AB - Arisaema serratum possesses a pitfall-trap flower pollination system. However, little is known about the efficiency and pattern of pollen movement in A. serratum. Thus, the aims of this study are to (1) determine the paternal parents of the seeds and (2) elucidate pollen movement in a natural population. Paternity analysis using microsatellite markers was performed. Seeds were collected from a natural population of A. serratum in 2001 at Horigane, Japan. Small midges became trapped in female spathe tubes during the flowering period. We found that (1) seeds in a fruit were fertilized by multiple sires; (2) seeds sired by a paternal parent were either clumped, exclusively, or randomly distributed on the spadix, depending on the parent; (3) to a great extent, a few males contributed as sires; (4) distance from a female was not a factor in the inequality of reproductive success among males; (5) male reproductive success was not correlated with its size. We conclude that pollen carryover and the trap-flower pollination system are likely to result in multiple paternity and inequality in male success. PMID- 21646134 TI - Testing for ecological and genetic Allee effects in the invasive shrub Senna didymobotrya (Fabaceae). AB - For an introduced plant species to become invasive, it must be able to reproduce even in initially small populations. We tested for Allee effects (reduced reproductive performance of individuals in small populations) in the nonclonal, buzz-pollinated shrub Senna didymobotrya in its invasive range in South Africa. The species is self-compatible, but we found that in its invasive range in South Africa it requires pollinators to set seed. Nearly all stigmas (90%) received pollen, but natural fruit set was very low (3-20%). Pollen receipt and fruit set were not significantly correlated with population size. We thus found no evidence for an ecological Allee effect arising from pollen limitation in small populations. Offspring seedling performance, measured in terms of stem volume and leaf area, was also not significantly correlated with the number of plants in the source population, indicating that genetic Allee effects, such as inbreeding depression, are either absent or of such a small magnitude that they would be unlikely to limit further spread of S. didymobotrya in South Africa. PMID- 21646135 TI - A Late Devonian isoetalean lycopsid, Otzinachsonia Beerboweri, gen. et sp. nov., from north-central Pennsylvania, USA. AB - Compressions and impressions of an isoetalean lycopsid, comprising lower portions of stems, lobed bases, attached rootlets, and rounded rootlet scars, discovered in Late Devonian (Famennian) rocks of Clinton County, north-central Pennsylvania, Appalachian Basin, USA, are here described as Otzinachsonia beerboweri, gen. et sp. nov. These specimens demonstrate unequivocally the existence of the isoetalean lobe-and-furrow rhizomorphic growth pattern as early as the Late Devonian. They were found in an Archaeopteris- and Rhacophyton-dominated flora at Red Hill, an outcrop of the Duncannon Member of the Catskill Formation. The fossils were found in a dark-gray to greenish-gray lenticular siltstone layer that has an average thickness of 1.0 m. This deposit is interpreted as a floodplain pond. The low-energy nature of the deposit and the fine preservation of the intact rootlets of the specimens imply little or no transport. The plants were probably growing along the edge of the floodplain pond with their lower portions submerged for at least part of the year. PMID- 21646136 TI - Correlations of climate and plant ecology to leaf size and shape: potential proxies for the fossil record. AB - The sizes and shapes (physiognomy) of fossil leaves are widely applied as proxies for paleoclimatic and paleoecological variables. However, significant improvements to leaf-margin analysis, used for nearly a century to reconstruct mean annual temperature (MAT), have been elusive; also, relationships between physiognomy and many leaf ecological variables have not been quantified. Using the recently developed technique of digital leaf physiognomy, correlations of leaf physiognomy to MAT, leaf mass per area, and nitrogen content are quantified for a set of test sites from North and Central America. Many physiognomic variables correlate significantly with MAT, indicating a coordinated, convergent evolutionary response of fewer teeth, smaller tooth area, and lower degree of blade dissection in warmer environments. In addition, tooth area correlates negatively with leaf mass per area and positively with nitrogen content. Multiple linear regressions based on a subset of variables produce more accurate MAT estimates than leaf-margin analysis (standard errors of +/-2 vs. +/-3 degrees C); improvements are greatest at sites with shallow water tables that are analogous to many fossil sites. The multivariate regressions remain robust even when based on one leaf per species, and the model most applicable to fossils shows no more signal degradation from leaf fragmentation than leaf-margin analysis. PMID- 21646137 TI - New insights into Paleozoic charophyte morphology and phylogeny. AB - Examination of Paleozoic charophyte fructifications using microscopy and high resolution x-ray synchrotron microtomography has revealed that most of them have a utricle that forms a supplementary calcified cover around the gyrogonite. All Paleozoic families with utricles have been assigned to the Sycidiales. We consider the Moellerinaceae to occupy a central position in the phylogeny of the Charophyta. From these, one group of descendants constitutes the gyrogonites inside the utricles of the Sycidiales; a second descendant goup is thought to be the utricle-free ancestors of the Charales prior to inversion of spiralization and reduction in cell number. The Sycidiales have a multilayered wall and an internal vesicle, but their families are distinguished by diversity in orientation of external cells, complexity of the utricle wall, and in presence or absence of antheridia. The solidly packed structure of the utricle is believed to be an organ protecting the zygote against dessication. We interpret the morphological similarities between Paleozoic Sycidiales and Mesozoic Clavatoraceae, both with a utricle, as homoplasous rather than expressing a true phylogenetic relationship. We confirm that some umbellids might correspond to utricles of charophytes. PMID- 21646138 TI - Genetic and clonal diversity of two cattail species, Typha latifolia and T. angustifolia (Typhaceae), from Ukraine. AB - Genetic and clonal diversity vary between two closely related cattail species (Typha angustifolia and T. latifolia) from Ukraine. This diversity was calculated from microsatellite data. Forty-eight percent of the total variation was partitioned between species, which formed distinct clusters in a dendrogram with no indication of hybrid populations. Typha angustifolia had higher heterozygosity at the species (H(es) = 0.66) and population (H(ep) = 0.49) levels than did T. latifolia (H(es) = 0.37 and H(ep) = 0.29, respectively). The higher number of alleles in T. angustifolia may be indicative of larger effective population sizes due to its higher seed production. Clonal diversity of T. angustifolia was lower than that of T. latifolia (N(g)/N(r) = 0.40 and 0.61, Simpson's D = 0.82 and 0.94, respectively). Correlations between clonal and genetic diversity were higher for T. latifolia than T. angustifolia, suggesting that the importance of factors and their interactions affecting this relationship are different for the two species. Latitudinal and longitudinal trends were not observed in either species despite the large sampling area. Population differentiation was relatively high with F(ST) of 0.24 and 0.29 for T. angustifolia and T. latifolia, respectively. Weak isolation by distance was observed for T. latifolia but not for T. angustifolia. PMID- 21646139 TI - Phylogenetics and diversification of Cotyledon (Crassulaceae) inferred from nuclear and chloroplast DNA sequence data. AB - Crassulaceae includes approximately 35 genera and 1500 species of leaf and stem succulent flowering plants. The family is nearly cosmopolitan in distribution, but is particularly diverse in southern Africa, where five genera comprising approximately 325 species are found. One of these genera, Cotyledon, includes 10 species that are largely confined to South Africa, where they are commonly found on rocky hillsides, coastal flats, and cliff faces. Species of Cotyledon are characterized by five-parted, pendulous, sympetalous flowers, but the genus is highly diverse in growth form, flower color and size, and leaf morphology. One particularly variable species, C. orbiculata, has been divided into five varieties based on leaf morphology and biogeography; however, the monophyly of this species as well as the relationships among the varieties have not previously been investigated. Parsimony analyses of a combined data set of DNA sequences from chloroplast and nuclear genome provided the first estimate of phylogeny for Cotyledon, and resulted in two minimum-length trees and a fully resolved phylogeny for the genus. Results indicate that C. orbiculata is not monophyletic and suggest the need for additional studies and a revised classification within the genus. PMID- 21646140 TI - Basal cactus phylogeny: implications of Pereskia (Cactaceae) paraphyly for the transition to the cactus life form. AB - The cacti are well-known desert plants, widely recognized by their specialized growth form and essentially leafless condition. Pereskia, a group of 17 species with regular leaf development and function, is generally viewed as representing the "ancestral cactus," although its placement within Cactaceae has remained uncertain. Here we present a new hypothesis of phylogenetic relationships at the base of the Cactaceae, inferred from DNA sequence data from five gene regions representing all three plant genomes. Our data support a basal split in Cactaceae between a clade of eight Pereskia species, centered around the Caribbean basin, and all other cacti. Two other Pereskia clades, distributed mostly in the southern half of South America, are part of a major clade comprising Maihuenia plus Cactoideae, and Opuntioideae. This result highlights several events in the early evolution of the cacti. First, during the transition to stem-based photosynthesis, the evolution of stem stomata and delayed bark formation preceded the evolution of the stem cortex into a specialized photosynthetic tissue system. Second, the basal split in cacti separates a northern from an initially southern cactus clade, and the major cactus lineages probably originated in southern or west-central South America. PMID- 21646141 TI - Molecular phylogenetics of Limonium and related genera (Plumbaginaceae): biogeographical and systematic implications. AB - Phylogenetic relationships within Limonium (Plumbaginaceae) are evaluated using sequence data from three plastid regions (rbcL, the trnL intron, and the trnL trnF intergenic spacer). Sixty-six species representing the major genera of Staticoideae, including representatives of all sections and genera formerly included in Limonium, have been analyzed using four species of Plumbaginoideae as an outgroup. Analyses of each separate and combined data set yield similar results. Afrolimon is embedded in Limonium and related to L. vulgare, the type of Limonium. Limonium is split into two major clades corresponding to subgenera, but otherwise the current infrageneric classification proved to be artificial. Some groups restricted to particular areas can be recognized, and their synapomorphies are discussed. The presence of an isolated taxon in the Canary Islands is used as a calibration point for age estimates of the major events in the genus, including migrations to the Southern Hemisphere, the Canary Islands, and Asia. The rapid radiation of Limonium in the Mediterranean basin appears to coincide with the desiccation of the Mediterranean Sea in the Messinian (late Miocene). PMID- 21646142 TI - Systematics and biogeography of Lathyrus (Leguminosae) based on internal transcribed spacer and cpDNA sequence data. AB - Lathyrus (Leguminosae; Papilionoideae) is the largest genus in tribe Fabeae and exhibits an intriguing extratropical distribution. We studied the systematics and biogeography of Lathyrus using sequence data, from accessions representing 53 species, for the internal transcribed spacer plus 5.8S-coding region of nuclear ribosomal DNA as well as the trnL-F and trnS-G regions of chloroplast DNA. Our results generally supported recent morphology-based classifications, resolving clades corresponding to sections Lathyrus and Lathyrostylis, but question the monophyly of the large, widespread section Orobus sensu Asmussen and Liston. Sections Orobus, Aphaca, and Pratensis form a predominantly northern Eurasian-New World clade. Within this clade, the North American and eastern Eurasian species, including both Holarctic species (L. palustris and L. japonicus), form a transberingian clade of relatively recent origin and diversification. The South American Notolathyrus group is distant from this transberingian lineage and should be reinstated as a distinct section within the northern Eurasian-New World clade. The Notolathyrus lineage reached the New World most probably through long distance dispersal from Eurasia. The remaining sections in the genus are centered on the Mediterranean region. PMID- 21646143 TI - Within-crown variation in the timing of leaf emergence and fall of Malaysian trees in association with crown development patterns. AB - In aseasonal tropics, timing of leaf emergence and leaf fall may differ between the shoots of different crown parts within a tree. This is important for the efficient development of crowns because leaves should be produced as soon as enough carbohydrates are accumulated. This hypothesis was tested by investigating leaf demography over a 44-mo period for 17 Malaysian trees and comparing the timings of leaf emergence and fall between the upper and lower crowns. The timings of leaf emergence were synchronized between the upper and lower crowns, but those of leaf fall were less synchronized in most trees. Greater rates of leaf production in the upper than in the lower crowns were attributable to the differences in the number of leaves that emerged per leaf emergence event, rather than differences in frequency of leaf emergence per year. Timings of leaf emergence and leaf fall were mainly simultaneous in the upper and lower crowns, but unsynchronized leaf production and leaf fall also occurred. Such limited plasticity of leaf demography within crowns may be the result of physiological integration of branches or the compromise between the advantages of satiating herbivores and effective crown development in the trees of aseasonal tropics. PMID- 21646144 TI - Codon usage patterns distort phylogenies from or of DNA sequences. AB - Papers reporting phylogenetic reconstructions often include discussion of the nature of third position substitutions and have often treated third position data differently from other data. This paper extends such considerations. Plant biotechnologists interested in high levels of expression of foreign proteins have accumulated information on preferences for otherwise synonymous codons. This paper presents a simple analysis for codon bias. Not only is bias frequent, but bias also varies between cohorts of proteins, both by amino acid and by taxon. Analysis of codon usage in the parallel divergence of phytochromes in three model plants finds identical bias for all family members within each taxon and increasingly divergent patterns of bias between increasingly divergent taxa. The molecular constraint of taxon-specific pools of tRNA molecules means individual triplets in a coding sequence are often not independent; algorithms designed to analyze independent characters are inappropriate for such data. Although a misestimate of the number of differences between taxa and groups of taxa can still generate an accurate description of the nesting of clades, other phylogenetic parameters will be strongly affected. Importantly, since codon bias produces smaller-than-expected within-taxon variance (common use of favored triplets) and larger-than-expected between-taxa variance (different favorites in different taxa), statistical support for nodes is certain to be wrong. The translational control of gene expression mediated by codon bias has implications for modern molecular systematics. PMID- 21646145 TI - Comparative infructescence morphology in Liquidambar (Altingiaceae) and its evolutionary significance. AB - The sweet gum genus Liquidambar (Altingiaceae) has two species in eastern Asia, one in eastern North America, and one in western Asia. Mature infructescences are studied to provide anatomical, morphological, and micromorphological details, some of which are newly recognized. Homology is suggested between extrafloral spinose processes of L. formosana and L. acalycina, braid-like ornamentation of L. styraciflua, and broad intercarpellate areas of L. orientalis. Morphology, position, number, and the presence of similar structures in the closely related Hamamelidaceae s.s. support their derivation from sterile flowers. Morphological cladistic analysis using 43 characters supports the monophyly of Liquidambar with Altingia as its sister. The matK analysis contrastingly places Altingia sister to the L. acalycina-L. formosana clade, rendering Liquidambar paraphyletic. Discordance between morphological and matK data sets may result from both different rates of morphological evolution and convergence. Several similarities between Altingia and L. acalycina are symplesiomorphic in the morphological cladistic analysis. Microaltingia apocarpela, from the Cretaceous of eastern North America, documents the earliest known fossil divergence within Altingiaceae. The Miocene Liquidambar changii of western North America is sister to a clade of extant Liquidambar species. Consideration of this fossil evidence reveals complex intercontinental biogeographic disjunctions in Altingiaceae. PMID- 21646146 TI - N, P, and C stoichiometry of Eranthis hyemalis (Ranunculaceae) and the allometry of plant growth. AB - We report the nitrogen (N), phosphorus (P), and carbon (C) stoichiometry for each of the five organ-types (leaves, aerial stems, reproductive organs, roots, and tubers) of 17 actively growing Eranthis hyemalis plants differing in size (as measured in g C). We also report the N, P, and C stoichiometry of 20 winterized tubers, which are the only perennial organs of this species. Comparisons between whole-plant and winterized N/C and P/C levels indicate that N was resorbed from aerial organs and stored in tubers by the end of the growing season. Leaves were substantial reservoirs for N and P. With few exceptions, N scaled isometrically with respect to C for each organ-type, whereas P scaled as the 3/4 power of C. Thus, N is proportional to P(3/4), which is proportional to C regardless of organ type. Additionally, annual growth rate G of shoots (leaves and aerial stems) scaled as the -3 power of leaf N/P quotients such that G was proportional to the 3/4 power of leaf P. We suggest that these scaling relationships (together with previously reported allometric trends across herbaceous species) show that growth is constrained by organ-specific N and P allocation patterns (presumably to proteins and ribosomes, respectively). PMID- 21646147 TI - Pollination success across an elevation and sex ratio gradient in gynodioecious Daphne laureola. AB - Gynodioecy is a dimorphic breeding system in which hermaphrodite and female individuals coexist in populations. Sex ratio and gender-relative lifetime seed production determine the stability of gynodioecy, and both genetic and ecological factors may influence these parameters. I analyzed the consequences of variation in population sex ratio and site elevation for the relative pollination success of female and hermaphrodite individuals of Daphne laureola in southern Spain, where previous studies failed to detect female fecundity advantages at two mid elevation sites. Pollination success, estimated as stigmatic pollen loads, number of pollen tubes per style, and percentage of fertilized flowers, was higher for hermaphrodites than females in populations with 20-56% females. Furthermore, female quantitative disadvantage in pollination success increased with elevation, suggesting that the higher availability of pollen due to the increased proportion of hermaphrodites could not mitigate the negative effect that other factors associated with elevation apparently had on pollination. Supplemental hand pollinations showed that female seed production was pollen limited in populations with a proportion of females >50%, although both pollination success and natural fruit set of females in these sites were the highest recorded. PMID- 21646148 TI - Nectar traits in Nicotiana section Alatae (Solanaceae) in relation to floral traits, pollinators, and mating system. AB - Nicotiana section Alatae exhibits great diversity among species in floral morphology, mating system, and predominant pollinators. As a first step towards estimating nectar's role in floral evolution, we studied nectar traits to determine whether they vary in association with predominant pollinators and mating system. Daily phenology determines when nectar becomes available to pollinators and differed between hummingbird- and moth-pollinated species. Nectar volume and concentration varied significantly among most species and pollinator groups, but were inversely correlated, so that total energy was similar among most species. In general, nectar volume was positively correlated with corolla length. The autogamous species, N. plumbaginifolia, had a nectar volume that matched expectations based on corolla length, but with lower concentration and total energy than predicted by corolla length, while nectar volume was lower than predicted by corolla length in the autogamous population of N. longiflora. Sugar and amino acid components (determined through HPLC) were similar among species, although differences did exist. The nectar of most species was sucrose-dominant, but the autogamous N. plumbaginifolia had nectar that contained similar proportions of sucrose, glucose, and fructose. Many nectar traits varied in association with the predominant pollinators and, in some cases, with the mating system. PMID- 21646149 TI - Duplication of floral regulatory genes in the Lamiales. AB - Duplication of some floral regulatory genes has occurred repeatedly in angiosperms, whereas others are thought to be single-copy in most lineages. We selected three genes that interact in a pathway regulating floral development conserved among higher tricolpates (LFY/FLO, UFO/FIM, and AP3/DEF) and screened for copy number among families of Lamiales that are closely related to the model species Antirrhinum majus. We show that two of three genes have duplicated at least twice in the Lamiales. Phylogenetic analyses of paralogs suggest that an ancient whole genome duplication shared among many families of Lamiales occurred after the ancestor of these families diverged from the lineage leading to Veronicaceae (including the single-copy species A. majus). Duplication is consistent with previous patterns among angiosperm lineages for AP3/DEF, but this is the first report of functional duplicate copies of LFY/FLO outside of tetraploid species. We propose Lamiales taxa will be good models for understanding mechanisms of duplicate gene preservation and how floral regulatory genes may contribute to morphological diversity. PMID- 21646150 TI - Welwitschiaceae from the Lower Cretaceous of northeastern Brazil. AB - Welwitschiaceae, a family in the Gnetales, is known today from only one extant species, Welwitschia mirabilis. This species is distributed in the Namibian desert, along the western coast of southern Africa, about 10 km inland from the coast. Very little is known about the fossil record of this family. Lower Cretaceous megafossils of various organs, assigned to Welwitschiaceae, are presented here. These fossils include young stems with paired cotyledons attached (Welwitschiella austroamericana n. gen. et sp.), isolated leaves (Welwitschiophyllum brasiliense n. gen. et sp.), and axes bearing male cones (Welwitschiostrobus murili n. gen. et sp.). They were collected in the Crato Formation, which is dated by palynomorphs and ostracods as Late Aptian (114 to 112 million years ago). These sediments are exposed in the Araripe Basin of northeastern Brazil. This study brings together new information of the megafossil record of Welwitschia-like plants and also reports of pollen said to be similar to that of Welwitschia from Lower Cretaceous sediments. PMID- 21646151 TI - Drought tolerance in the alpine dandelion, Taraxacum ceratophorum (Asteraceae), its exotic congener T. officinale, and interspecific hybrids under natural and experimental conditions. AB - We compared water relations and adaptations to drought stress in native and invasive exotic dandelions, Taraxacum ceratophorum and T. officinale. Photosynthesis (A), transpiration (E), and water use efficiency (WUE; carbon gained/water lost) were measured for the two species under extreme drought in the alpine tundra of Colorado, USA. We also subjected both species and F(1) hybrids to a dry-down experiment to determine how relative physiological performance varied with water availability. Photosynthesis and transpiration in the field were low and did not differ between Taraxacum congeners; however, native T. ceratophorum had higher WUE than T. officinale. After 6 days of greenhouse drought, photosynthesis and transpiration were reduced in T. officinale compared to T. ceratophorum. Taraxacum ceratophorum maintained high WUE under control and drought treatments. Conversely, WUE in T. officinale was highly plastic between watered (low WUE) and dry-down (high WUE) treatments. Hybrids did not exhibit heterosis; instead, they were similar to T. officinale in A and E and intermediate to the parental species in WUE. Overall, results suggest that native dandelions are more drought tolerant than invasive congeners or their hybrids, but have less plasticity in WUE. Arid habitats and occasional drought in mesic sites may provide native dandelions with refugia from negative interactions with invasives. PMID- 21646152 TI - Testing for stress-dependent inbreeding depression in Impatiens capensis (Balsaminaceae). AB - The relevance of inbreeding depression to the persistence of plant populations can depend upon whether stress magnifies inbreeding depression for fitness related traits. To examine whether drought stress exacerbates inbreeding depression in gas exchange traits and biomass, we grew selfed and outcrossed progeny of inbred lines from two populations of Impatiens capensis in a greenhouse experiment under water-limited and moist soil conditions. Drought stress did not magnify the degree of inbreeding depression for any of the traits measured. In fact, in one population there was a trend for stronger inbreeding depression under well-watered, benign conditions. Furthermore, significant inbreeding depression for carbon assimilation rate and stomatal conductance was only detected in the lines from one population. In contrast, inbreeding depression for biomass was detected within both populations and differed among lines. Drought stress exerted significant selection on physiological traits, favoring increased carbon assimilation rates and decreased stomatal conductance in drought-stressed plants. Patterns of selection did not differ between inbred and outcrossed plants but did differ marginally between populations. Thus, estimates of selection were not biased by the mixed mating system per se, but may be biased by combining individuals from populations with different histories of selection and inbreeding. PMID- 21646153 TI - Reproductive ecology of five sympatric Agave Littaea (Agavaceae) species in central Mexico. AB - We studied a community of Agave species that coexist in the Metztitlan Canyon in Central Mexico. During 2001, 2002, and 2003, we analyzed floral traits and rosette allometry in five species belonging to the subgenus Littaea: A. celsii albicans, A. xylonacantha, A. difformis, A. striata, and Agave sp.; and observed floral visitors for each species. We report the first evidence of bat visitation in the subgenus Littaea and find that bats (Leptonycteris curasoae, Choeronycteris mexicana, and Glossophaga sp.) are the primary pollinators in four species. Honeybees, hummingbirds, bumblebees and hawkmoths were also common visitors. We propose that the presence of diurnal pollinators may increase the reproductive success of the plant when offering pollinator services additional to the coadapted pollinator. We also found evidence of selection pressures toward semelparity because pollinators are selecting for taller inflorescences in three of the five species. There is phenological complementarity in this community because the flowering periods of the five species span the entire year, although there are some periods when pairs of species overlap. Additionally, we found evidence for character displacement in rosette sizes and separation of spatial and temporal resource use in pollinator composition among species. PMID- 21646154 TI - Exploitation of a specialized mutualism by a deceptive orchid. AB - Plants that lack floral rewards may nevertheless attract pollinators through mimetic resemblance to the flowers of co-occurring rewarding plants. We show how a deceptive orchid (Disa nivea) successfully exploits a reciprocally specialized mutualism between a nectar-producing plant (Zaluzianskya microsiphon) and its long-proboscid fly pollinator (Prosoeca ganglbaueri). Disa nivea is a rare southern African orchid known only from habitats that support large populations of Z. microsiphon, which it closely resembles in both general morphology and floral spectral reflectance. Significant covariation in floral traits of Z. microsiphon and D. nivea was detected among populations. Where mimics are uncommon, flies do not appear to discriminate between the flowers of the two species. Pollination success in D. nivea was much higher at a site with abundant Z. microsiphon plants than at a site where Z. microsiphon was rare. Exploitation of a highly specialized mutualism appears to demand a high degree of phenotypic resemblance to a rewarding model by a deceptive mimic, as exemplified by D. nivea. The majority of deceptive orchids, on the other hand, exploit relatively generalized pollination systems and thus require only a vague resemblance to rewarding plants in the community in order to attract pollinators. PMID- 21646155 TI - Development and structure of trichotomous branching in Edgeworthia chrysantha (Thymelaeaceae). AB - We studied the development and structure of the unusual trichotomous branching of Edgeworthia chrysantha. Three "branch primordia" are formed sequentially on the shoot apex of a main axis and develop into trichotomous branching. The branch primordia are clearly distinguishable from the typical axillary buds of other angiosperms; they develop much more rapidly than axillary buds, and the borders between the branch primordia and shoot apex of the main axis are anatomically unclear. Furthermore, at a later stage, leaves subtending the branch primordia produce typical axillary buds. These results suggest that the trichotomous branching in this species involves the division of the shoot apical meristem. Expression analysis of genes involved in branching or maintenance of the shoot apical meristem in this species should clarify the control mechanism of this novel branching pattern in angiosperms. We also observed the phyllotactic patterns in trichotomous branching and have related these patterns to the shoot system as a whole. PMID- 21646156 TI - A phylogenetic reappraisal of the Peltophorum group (Caesalpinieae: Leguminosae) based on the chloroplast trnL-F, rbcL and rps16 sequence data. AB - The monophyly of the Peltophorum group, one of nine informal groups recognized by Polhill in the Caesalpinieae, was tested using sequence data from the trnL-F, rbcL, and rps16 regions of the chloroplast genome. Exemplars were included from all 16 genera of the Peltophorum group, and from 15 genera representing seven of the other eight informal groups in the tribe. The data were analyzed separately and in combined analyses using parsimony and Bayesian methods. The analysis method had little effect on the topology of well-supported relationships. The molecular data recovered a generally well-supported phylogeny with many intergeneric relationships resolved. Results show that the Peltophorum group as currently delimited is polyphyletic, but that eight genera plus one undescribed genus form a core Peltophorum group, which is referred to here as the Peltophorum group sensu stricto. These genera are Bussea, Conzattia, Colvillea, Delonix, Heteroflorum (inedit.), Lemuropisum, Parkinsonia, Peltophorum, and Schizolobium. The remaining eight genera of the Peltophorum group s.l. are distributed across the Caesalpinieae. Morphological support for the redelimited Peltophorum group and the other recovered clades was assessed, and no unique synapomorphy was found for the Peltophorum group s.s. A proposal for the reclassification of the Peltophorum group s.l. is presented. PMID- 21646157 TI - Speciation and hybridization among Houstonia (Rubiaceae) species: the influence of polyploidy on reticulate evolution. AB - Chloroplast and nuclear DNA sequence variation among populations and species was used to examine the phylogenetic history and hybridization of the North American Houstonia lineage. The ancestral species in the lineage do not show evidence of hybridization; however, the more recently derived species in eastern North America contain a wide degree of morphological and genetic variation both within and among species. Furthermore, there is a clear association between hybridization and polyploidy in the Houstonia lineage, with all potential hybrids occurring among species that contain polyploid populations. This suggests that polyploidy may break down species barriers and allow hybridization among lineages. These results indirectly support speciation models that involve genetic incompatibilities among species arising from gene silencing or genomic reorganization. PMID- 21646158 TI - Evolution of shrub-like growth forms in the lianoid subfamily Secamonoideae (Apocynaceae s.l.) of Madagascar: phylogeny, biomechanics, and development. AB - Lianas are common in the Apocynaceae s.l. and are predominant in the subfamily Secamonoideae. Shrub-like taxa are rare within this subfamily but occur in Malagasy genera such as Secamone, Secamonopsis, and Pervillaea. We explored the evolutionary appearance of shrub-like growth forms in Malagasy Secamonoideae through a molecular phylogeny using chloroplastic sequences. The phylogeny revealed several independent appearances of shrub-like growth forms within the Secamonoideae. Biomechanics and development of the shrub-like growth form were detailed in one species, Secamone sparsiflora, which has upright and self supporting young stems that become procumbent in older stages of development. Biomechanical investigations revealed characteristics atypical of both lianas and self-supporting shrubs. Anatomical development in S. sparsiflora is initially similar to lianas in the same clade but shows potentially neotenic retention of juvenile wood development for most of the growth trajectory. The results suggest that evolution of lianescence can carry a high degree of specialization and developmental burden that might limit evolution back to self-supporting growth forms. Under certain geographic and ecological conditions, such as geographic isolation, xeric conditions and/or reduced biotic competition, escapes from lianescence to other growth forms can occur in some angiosperm groups via relatively simple heterochronic shifts of mechanically significant growth processes. PMID- 21646159 TI - Molecular phylogenetic analysis of uniovulate Euphorbiaceae (Euphorbiaceae sensu stricto) using plastid RBCL and TRNL-F DNA sequences. AB - Parsimony and Bayesian analyses of plastid rbcL and trnL-F DNA sequence data of the pantropical family Euphorbiaceae sensu stricto (s.s.) are presented. Sampling includes representatives of all three subfamilies (Acalyphoideae, Crotonoideae, and Euphorbioideae), 35 of 37 tribes and 179 of the 247 genera of uniovulate Euphorbiaceae sensu lato (s.l.). Euphorbiaceae s.s. were recovered as a monophyletic group with no new adjustments in circumscription. Two clades containing taxa previously placed in Acalyphoideae are found to be successive sisters to all other Euphorbiaceae s.s. and are proposed here at subfamilial rank as Peroideae and Cheilosoideae. The remainder of the family fall into seven major lineages including Erismantheae and Acalyphoideae s.s. (parts of Acalyphoideae), Adenoclineae s.l., Gelonieae, articulated crotonoids and inaperturate crotonoids (parts of Crotonoideae), and Euphorbioideae. Potential synapomorphies and biogeographical trends are suggested for these clades. Acalyphoideae s.s., inaperturate crotonoids, and Euphorbioideae tribe Hippomaneae each have two major subclades that represent novel groupings without apparent morphological synapomorphies. Two subfamilies, 14 tribes, and 10 genera were found to be para- or polyphyletic. Noteworthy among these, Omphaleae are embedded in Adenoclineae, Hureae + Pachystromateae in Hippomaneae, Ditta in Tetrorchidium, and Sapium s.s. in Stillingia. PMID- 21646160 TI - Leaf architecture and direction of incident light influence mesophyll fluorescence profiles. AB - Light propagation and distribution inside leaves have been recognized as important processes influencing photosynthesis. Monochromatic light absorption across the mesophyll was measured using chlorophyll fluorescence generated from illumination of the cut edge (epi-illumination), as well as the adaxial or abaxial surfaces of the leaf. Species were selected that had basic leaf types: laminar leaf with adaxial palisade layer (Rhododendron catawbiense), needle with palisade (Abies fraseri), and needle without palisade (Picea rubens). Fluorescence was more evenly distributed across the mesophyll for adaxially illuminated leaves with a palisade cell layer, as well as for the needles (cylindrical) without palisade, when compared to fluorescence generated by abaxial illumination. Moreover, fluorescence from green light illumination remained high across the mesophyll of adaxially illuminated R. catawbiense, indicating a possible influence of mesophyll structure on internal light distribution beyond that of chlorophyll levels. These data support the idea that light propagation within the mesophyll is associated with asymmetric mesophyll structure, in particular the presence of palisade cell layers. In addition, we propose that the evolution of a more cylindrical leaf form, such as found in conifer species, may be a structural solution to excessive sunlight that replaces the highly differentiated mesophyll found in most laminar-leaved species. PMID- 21646161 TI - Evolution of reproductive structures in grasses (Poaceae) inferred by sister group comparison with their putative closest living relatives, Ecdeiocoleaceae. AB - Despite much recent activity in the phylogeny and developmental genetics of grasses, the enigmatic homologies of their reproductive structures remain largely unresolved, partly because their highly derived morphology has resulted in a unique associated terminology. Outstanding questions include whether grass lodicules and stamens are derived from a single perianth or stamen whorl, respectively, whether the grass caryopsis is homologous with a nut, and how the scutellum evolved. We investigated the reproductive structures of the putative sister group of grasses, the southwestern Australian family Ecdeiocoleaceae, which includes two genera, Ecdeiocolea and Georgeantha. The zygomorphic arrangement of the four (rather than six) stamens in male flowers of Ecdeiocolea indicates that they may represent three outer stamens plus the adaxial inner stamen. Within Ecdeiocoleaceae, characters such as the highly unusual nucellus structure of Ecdeiocolea are autapomorphic. Sister-group comparisons indicate that some characteristic grass features, notably the scutellum, do not occur in their putative closest relatives and that more data are needed on early-diverging grass genera to resolve these issues. The grass caryopsis could represent one end of a transformation series embodied by the reduced gynoecial structure and indehiscent fruit of other Poales such as Flagellaria, Joinvillea, and Ecdeiocolea. PMID- 21646162 TI - Biomechanics and development of the climbing habit in two species of the South American palm genus Desmoncus (Arecaceae). AB - Mechanical properties are investigated in Desmoncus orthacanthos and D. polyacanthos from French Guiana, South America. Differences in size and axis stiffness are related to different trellis requirements and habitats. The leaf sheath surrounds the stem, increasing stiffness of young self-supporting stages and apical parts of older climbing plants. Senescence of the leaf sheath reduces stiffness of older climbing axes of both species. Its eventual loss in D. orthacanthos facilitates deformation into coils and loops when plants slip from their supports following senescence of leaves bearing attachment organs. In smaller climbing axes of D. polyacanthos, the senescent leaf sheath remains attached and axes rarely form loops and coils below attachment. An increase in stiff mechanical properties toward the base of both species is radically different from that of many dicotyledonous lianas. Besides the presence of attachment organs, stem mechanical properties of Desmoncus are similar to those of erect though not fully self-supporting stems of Bactris major, a sympatric species of the sister group genus to Desmoncus. The climbing habit in Desmoncus may have evolved via (1) heterochronic processes including early elongation of internodes relative to increase in stem diameter (reduction of the establishment phase) and (2) increased persistence of leaf sheaths. PMID- 21646163 TI - Growth and nutrient responses of Eloecharis cellulosa (Cyperaceae) to phosphate level and redox intensity. AB - Phosphorus (P) availability limits plant growth in many ecosystems. The ability of plants to explore for soil P is often impaired by nonresource stressors. Understanding the effects of these stressors on P acquisition in oligotrophic environments is critical in predicting species dominance. Growth and nutrient responses of Eleocharis cellulosa to redox intensity and phosphate level were evaluated under three redox potentials (Eh) and three phosphate (PO(4)) levels (P). Although low Eh (-150 mV) decreased root length at low P, Eh did not affect shoot height, relative growth rate (RGR), shoot elongation, photosynthesis, or biomass of E. cellulosa. Low PO(4) (10 MUg P . L(-1)) strongly inhibited growth. Shoot height, RGR, elongation, photosynthesis, and biomass were lower at 10 MUg P . L(-1) than at 80 or 500 MUg P . L(-1). None of the growth variables, except the ratio of root-supported biomass to root biomass, significantly differed between the 80 and 500 MUg P . L(-1) treatments. At low P, plants allocated relatively more biomass to roots than to shoots, compared to the medium and high P levels. Eleocharis cellulosa is well adapted to flooded conditions that lower soil Eh, and elevated PO(4) levels further promote its growth potential. PMID- 21646164 TI - Reproductive fitness of hybrids between Senecio jacobaea and S. aquaticus (Asteraceae). AB - Natural hybridization is increasingly recognized as an important process for the ecology and evolution of natural plant populations and species. There is a great need to initiate more studies based on natural populations in order to elucidate the possible role of hybrids in nature. The reproductive success of early generation hybrids can make or break hybrid lineages and may determine the genetic structure of hybrid swarms or the potential for gene flow through future generations, but studies of hybrid reproductive success are lacking. Here we measured components of male and female reproductive success in Senecio jacobaea and S. aquaticus (Asteraceae) species and F(1) hybrids between these species under laboratory conditions, and we measured reproductive output from crosses producing F(1), F(2), and backcross (BC) generation hybrids. F(1) hybrids were readily produced, and on average, the success of crosses producing subsequent generations (F(2), BC) decreased (though remained substantial), but the success of crosses was highly dependent on the genotypes involved. Also, F(1) hybrids were bigger, produced more flowers, and therefore produced more pollen than parental plants. Finally, crosses between parents were asymmetrical, such that S. aquaticus produced more and larger F(1) seeds than did S. jacobaea. PMID- 21646165 TI - An extinct calycanthoid taxon, Jerseyanthus calycanthoides , from the Late Cretaceous of New Jersey. AB - A new fossil genus and species, Jerseyanthus calycanthoides, is described from the Late Cretaceous (Turonian, ~90 MYBP) Raritan Formation of New Jersey. Flowers have cupulate receptacles bearing imbricately arranged tepals that subtend a series of recurved tepals near the cup margin. Recurved tepal subtends a "stamen staminode" pair, that includes a laminar stamen with ramified connective extensions, and an outer staminode. Outer staminodes are geniculate and incurved, and in aggregate their inner extremities define a circular area above the carpels and carpellodes. Each "stamen-staminode" pair apparently subtends (is opposite to) an inner tepal. Pollen is rounded and disulculate, with tectate columellate wall structure. Carpels are located at the center of the receptacle and have elongate styles that extend to and beyond the opening defined by the staminodal organs. Carpels are surrounded by tomentose carpellodes. Carpels include one marginally ridged seed. While these fossils do not match exactly any living species in morphology, they share numerous characters with extant members of Calycanthaceae and can be unequivocally placed within that family. Affinities of Jerseyanthus and Virginianthus were evaluated by including them in a combined analysis for the Laurales. Jerseyanthus is placed within Calycanthaceae as a sister taxon to the modern genus Calycanthus. PMID- 21646166 TI - Post-floral perianth functionality: contribution of persistent sepals to seed development in Helleborus foetidus (Ranunculaceae). AB - The perianth persists after flowering in many plants, yet its post-floral functionality has been little investigated, and the few studies available provide ambiguous evidence. This paper tests the hypothesis that the green persistent sepals of the perennial herb, Helleborus foetidus, contribute to the plant's fitness by enhancing seed number and/or size. Post-floral contribution of the calyx to fruit and seed development was evaluated by manipulating sepal number and measuring the effect on follicle size, seed set, seeds per follicle, and mean seed mass. The allometric relationship between calyx size and follicle mass was examined for flower buds, open flowers, and immature fruits differing in follicle number. Calyx manipulation had no significant effect on follicle size, seed set, or number of seeds per follicle, but it did have a significant influence on the size of individual seeds. Calyx size and mean seed mass were positively, linearly related. The calyx mass/seed number ratio declined with increasing number of follicles per fruit. The persistent sepals of Helleborus do contribute resources to the development of seeds, although there is not evidence of post-floral allometric adjustment of the calyx mass/follicle number ratio that could compensate for variations in seed number per fruit. PMID- 21646167 TI - Wind pollination, sexual dimorphism, and changes in floral traits of Schiedea (Caryophyllaceae). AB - Both changes in sex allocation and pollination mode may promote the separation of sexes in plant populations. Simultaneous evolution of wind pollination and dimorphism has occurred in Schiedea, where species with different female frequencies provide an opportunity to observe the effect of wind pollination on sex allocation and floral morphology. Differences among species in the ratio of anther to ovary volume were not the result of sex allocation trade-offs, but instead resulted from production of vestigial stamens in females; there were no changes in ovary volume in males and hermaphrodites (MH) of dimorphic species. Relative to hermaphroditic species, dimorphic species had more condensed inflorescences, a pattern often associated with wind pollination. Within dimorphic species, MH had longer filament lengths than females, and females had longer stigmas than MHs. These traits are characteristic of wind pollination, but there was no relationship between the degree of sexual dimorphism and female frequency. Ovary volume and ovule number and size had positive phenotypic correlations between females and MH of dimorphic species, making sex specialization more difficult. In dimorphic Schiedea species, selection for wind pollination may have a greater effect on floral traits than trade-offs in allocation between male and female function. PMID- 21646168 TI - The evolution of self-compatibility in geographically peripheral populations of Leavenworthia alabamica (Brassicaceae). AB - Self-compatibility and adaptations to self-fertilization are often found in plant populations at the periphery of species' ranges or on islands. Self-compatibility may predominate in these environments because it provides reproductive assurance when pollinators or availability of mates limits seed production. This possibility was studied in Leavenworthia alabamica, a flowering plant endemic to the southeastern United States. Populations at the center of the species' range retain sporophytic self-incompatibility, but peripheral populations are smaller, self-compatible, and have adaptations for self-fertilization. A reciprocal transplant experiment was designed to test whether there is pollen limitation of seed set and to examine its strength in central and peripheral populations. Self compatible genotypes produced more fruit and 17-22% more seed than self incompatible genotypes in all environments, suggesting that the transition to self-compatibility may be favored by natural selection in all populations inhabited by L. alabamica. Sequence analyses demonstrated that two peripheral populations have 90-100% reductions in genetic variation, consistent with the effects of small population size or historical bottlenecks. Although pollen limitation of seed set occurs in all environments, self-compatibility may evolve at the periphery in L. alabamica because the benefits of reproductive assurance are influenced by population size or bottlenecks following extinction and colonization. PMID- 21646169 TI - High abundance of dioecious plants in a tropical coastal vegetation. AB - We examined the frequency of hermaphroditic, monoecious, and dioecious species of angiosperms in restinga (sandy coastal plain) vegetation in SE Brazil. The study site was a vegetation mosaic comprising nine plant formations, ranging from open types to forest. Dioecy (14% of 566 species) was similar to other tropical vegetations and strongly associated with woodiness and entomophily. However, more interestingly, there was an exceptionally high percentage (35%) of dioecious species among the dominant woody plants. This pattern has not been previously reported, and we discuss the extent to which it is ecologically driven. We argue that the high abundance of dioecious plants in this resource-poor environment can be attributed to ecological traits related to long-distance dispersal, ecological vigor, and possibly, vegetative reproduction. PMID- 21646170 TI - Molecular phylogenetics of the giant genus Croton and tribe Crotoneae (Euphorbiaceae sensu stricto) using ITS and TRNL-TRNF DNA sequence data. AB - Parsimony, likelihood, and Bayesian analyses of nuclear ITS and plastid trnL-F DNA sequence data are presented for the giant genus Croton (Euphorbiaceae s.s.) and related taxa. Sampling comprises 88 taxa, including 78 of the estimated 1223 species and 29 of the 40 sections previously recognized of Croton. It also includes the satellite genus Moacroton and genera formerly placed in tribe Crotoneae. Croton and all sampled segregate genera form a monophyletic group sister to Brasiliocroton, with the exception of Croton sect. Astraea, which is reinstated to the genus Astraea. A small clade including Moacroton, Croton alabamensis, and C. olivaceus is sister to all other Croton species sampled. The remaining Croton species fall into three major clades. One of these is entirely New World, corresponding to sections Cyclostigma, Cascarilla, and Velamea sensu Webster. The second is entirely Old World and is sister to a third, also entirely New World clade, which is composed of at least 13 of Webster's sections of Croton. This study establishes a phylogenetic framework for future studies in the hyper-diverse genus Croton, indicates a New World origin for the genus, and will soon be used to evaluate wood anatomical, cytological, and morphological data in the Crotoneae tribe. PMID- 21646171 TI - Nuclear DNA, chloroplast DNA, and ploidy analysis clarified biological complexity of the Vandenboschia radicans complex (Hymenophyllaceae) in Japan and adjacent areas. AB - Species complexes consisting of ill-defined "species" are widely known among ferns, and their involvement with reticulate evolution is expected. Nevertheless approaches to reticulation history with DNA markers are not yet commonly adopted. We have successfully elucidated the biological status of the Vandenboschia radicans complex in East Asian islands by combining analyses of ploidy level, a cpDNA marker (rbcL), and a nuclear DNA marker (GapCp). The results based on 266 individuals collected from 174 localities throughout Japan and Taiwan suggest that complicated hybridizations have occurred involving at least three parental diploid species from within the V. radicans complex and Vandenboschia liukiuensis, which was formerly considered to be distinct from this complex. Triploids are the most common cytotype, but they show no evidence of apogamous reproduction, while all nonhybrid diploids are rare and have very limited distribution. Possible accounts of this phenomenon will be briefly discussed including the possibility of relict distribution and occasional apogamous reproduction. PMID- 21646172 TI - Molecular phylogeny of Oryzeae (Poaceae) based on DNA sequences from chloroplast, mitochondrial, and nuclear genomes. AB - The phylogeny and evolutionary history of the rice tribe (Oryzeae) were explored using sequences of five DNA fragments (matK, trnL, nad1, Adh2, and GPA1) from chloroplast, mitochondrial, and nuclear genomes. Results indicate that (1) Oryzeae is monophyletic and falls into two main clades corresponding to the traditionally recognized subtribes; (2) previous recognition of three monotypic genera (Hydrochloa, Porteresia, and Prosphytochloa) is not justified; and (3) close affinities of the monoecious genera are not supported, suggesting the possibility of multiple origins of unisexual florets. Based on the magnitude of matK and GPA1 sequence divergence, we suggest that Oryza and Leersia branched off from the remaining genera of Oryzeae ~20 million years ago (mya), and separated from each other ~14 mya. A divergence time of ~9 mya is obtained for the most basal split within Oryza. These estimates suggest that Oryzeae diverged during the Miocene, and thus imply that long-distance dispersal appears to be one of the important factors in the diversification of the tribe. PMID- 21646173 TI - Genetic variation is not concordant with morphological variation in the fern Asplenium hookerianum sensu lato (Aspleniaceae). AB - Species status cannot be adequately determined when partitions are based on only a single morphological character. For instance, the sympatry of plants with broad and narrow pinnules in the fern Asplenium hookerianum sensu lato from New Zealand creates the impression that two entities are present. The narrow-pinnuled plants are sometimes segregated as a distinct species, A. colensoi. However, this variation in pinnule morphology could equally be infraspecific, and only additional data can resolve this uncertainty. Analyses using AFLP DNA fingerprinting and DNA sequencing of the chloroplast trnL-trnF region indicate that genetic variation in A. hookerianum sensu lato is not concordant with pinnule morphology. Consequently, the recognition of A. colensoi is not supported. PMID- 21646174 TI - Life history and systematics of the aquatic discomycete Mitrula (Helotiales, Ascomycota) based on cultural, morphological, and molecular studies. AB - Mitrula species represent a group of aquatic discomycetes with uncertain position in the Helotiales and an unknown life history. Mitrula species were studied using a combination of cultural, morphological, and molecular techniques. Pure colonies were isolated from Mitrula elegans, and conidia were induced in vitro. Herbarium materials from Europe, Asia, and North America were studied. Sequences of rDNA, including partial small subunit rDNA, large subunit DNA and ITS, were used to infer phylogenetic relationships both within Mitrula and between Mitrula and other inoperculate discomycetes, with special attention to fungi that resemble Mitrula in morphology or ecology. Equally weighted parsimony analyses, likelihood analyses, constrained parsimony analyses, and Bayesian analyses were performed. Results suggest that (1) the anamorph of M. elegans produces brown bicellular conidia, (2) a new subalpine species M. brevispora is distinct, (3) more than six lineages and clades can be recognized in Mitrula, (4) the morphological species M. elegans is not monophyletic, (5) a close relationship between Mitrula and either Geoglossaceae or Sclerotiniaceae is not supported, (6) the Helotiaceae is paraphyletic, and (7) Mitrula belongs to a clade within the Helotiales that also includes other aero-aquatic genera, Cudoniella, Hydrocina, Vibrissea, Ombrophila, and Hymenoscyphus. PMID- 21646175 TI - The evolution and expression of RBCL in holoparasitic sister-genera Harveya and Hyobanche (Orobanchaceae). AB - The evolution of holoparasitism decreases the adaptive value of genes maintaining the photosynthetic apparatus. These may become pseudogenes through insertion or deletion events resulting in frameshift mutations, or by the evolution of premature stop codons. The holoparasitic sister genera Harveya and Hyobanche have undergone alternate pathways of evolution and expression at the plastid locus rbcL. An open reading frame in all but a single species of Harveya is maintained by purifying selection and is expressed. However, the function of Rubisco in this putative holoparasite is unknown. Conversely, Hyobanche has undergone rbcL pseudogene formation, and comparison of synonymous and nonsynonymous rates of evolution indicates that selection has not played a role in its evolution. This is complicated by the following findings: multiple pseudogene copies of rbcL exist in tissues of Hyobanche, rbcL transcripts also encode pseudogenes, and the large subunit is present in some tissues of Hyobanche. We hypothesize that the rbcL operon is in a state of degradation as may be expected in a holoparasite and is not endogenously expressed. Rather, the large subunit may be taken up from the host plants, and accumulate in tissues as a result of transpiration. PMID- 21646176 TI - Juvenile growth and palatability in co-occurring, congeneric British herbs. AB - Differential sensitivity (DS) storage dynamics describe a temporal niche axis that determines coexistence of competing taxa through a trade-off between environmental insensitivity and competitive ability at the recruitment stage. In DS storage dynamics, when the relevant environmental factor is low, the more sensitive, better competitor preferentially recruits; when the environmental factor is high, the environmentally sensitive species suffers high mortality and the environmentally insensitive taxon preferentially recruits. A herbivore defense/growth rate trade-off at the seedling/juvenile stage could support this dynamic. We therefore compared juvenile palatability, a measure of anti-herbivore defense, and early growth rate for five congeneric pairs of native British herbs. All five comparisons showed a positive association between average individual growth rate and average palatability to a native slug species. We observed no evidence of associations between early growth rate and adult palatability or between early growth rate and life history strategy (annual vs. perennial). Seed mass was not associated with either early growth rate or with life history strategy whether or not relatedness was taken into account. We offer two explanations as to why we found statistically significant support for a growth rate- defense trade-off when within-species studies so often produce only equivocal results. PMID- 21646177 TI - Developmental programmed cell death in primary roots of Sonoran Desert Cactaceae. AB - Primary roots of two species of Sonoran Desert Cactaceae, Stenocereus gummosus and Pachycereus pringlei, have a determinate pattern of growth: meristematic cells divide only for a limited time and then differentiate. Detecting DNA fragmentation by terminal deoxynucleotide transferase-mediated dUTP nick-end labeling (TUNEL), we have shown that programmed cell death (PCD) was not involved in meristem exhaustion. However, we found TUNEL-positive nuclei in the root hair and root cap cells of both species. Programmed cell death in root hair cells has not been previously reported, and the pattern of PCD events in the root cap differed from that described earlier. These data suggest that in the studied Cactaceae, PCD is involved in developmental adaptations related to the formation of a compact root system important for rapid seedling establishment in a desert environment. Participation of PCD in developmental loss of the root cap and in root hair renovation proposed in the current study implicates an evolutionary conserved link between PCD and differentiation processes in plants. PMID- 21646178 TI - A synchronous increase in hydraulic conductive capacity and mechanical support in conifers with relatively uniform xylem structure. AB - The dual function provided by longitudinal tracheids in conifers has led to a generally held trade-off concept that increasing wall thickness and/or volume of latewood tracheids improves mechanical support, while increasing cell diameter and/or volume of earlywood tracheids enhances conductive potential. Yet, some conifers have either uniform cell structure across the growth ring or, at most, a small amount of latewood. How do these trees accomplish the needs for increasing support and conduction with height growth? We examined Metasequoia glyptostroboides, a species that we previously demonstrated improves its mechanical properties with increasing age without a change in specific gravity or secondary wall microfibril angle. In this paper, we showed that lignin and extractive contents are not contributing factors, and through composite structure analysis, we eliminated a role for tracheid length. Using micromorphometric analysis, we demonstrated that as cell diameter increases, total primary wall decreases, secondary wall increases, and strength and conductive capacity increase with no change in specific gravity. Meta-analysis using other species of Cupressaceae, Podocarpaceae, and Araucariaceae provided strong corroborative evidence for this design strategy. PMID- 21646179 TI - Traits of invasives reconsidered: phenotypic comparisons of introduced invasive and introduced noninvasive plant species within two closely related clades. AB - In attempting to determine the traits associated with invasive plant species, ecologists have often used species native to the invaded range as "control species." Because many native species themselves are aggressive colonizers, comparisons using this type of control do not necessarily yield information relevant to distinctions between invasive and noninvasive species. Here we implement an alternative study design that compares phenological, architectural, size, and fitness traits of several introduced invasive species to introduced noninvasive species within two genera of Asteraceae (Crepis and Centaurea). While there were many significant differences between the genera, there were few shared attributes among invasive or noninvasive congeners, even for traits as seemingly important as the number of inflorescences produced and the size of seed heads. Instead, the results suggest that differences in invasiveness between closely related species is better explained as the result of complex trait interactions and specific introduction histories. PMID- 21646180 TI - Pinus baileyi (section Pinus, Pinaceae) from the Paleogene of Idaho, USA. AB - Pinus baileyi from the Paleogene of Idaho was initially related to the bristlecone pine P. longaeva (subgen. Strobus, sect. Parrya, subsect. Balfourianae) from western North America. Unlike the centromucronate condition in P. longaeva, P. baileyi cones have raised umbos that are excentromucronate, i.e., the mucro positioned in the upper umbo field above the keel. Cone size and scale morphology shows that P. baileyi more closely resembles excentromucronate pines of subsects. Halepenses and Pinus sensu Gernandt et al. (2005, Taxon 54: 29-42), but is most similar to P. resinosa, P. kesiya, and P. massoniana of subsect. Pinus. Morphologically, P. baileyi resembles the fossil species P. princetonensis and P. arnoldii from the Eocene Princeton Chert, British Columbia, Canada. Pinus baileyi extends the western North American range of ovulate cones resembling subsect. Pinus from the middle Eocene of British Columbia, Canada and Washington, USA to the Oligocene of Idaho, USA. Pinus baileyi, and possibly P. princetonensis and P. arnoldii, indicates the presence of early populations of subsect. Pinus type pines in the western cordillera of North America, raising the possibility that P. resinosa and P. tropicalis may have evolved from this group. PMID- 21646181 TI - High interpopulation genetic differentiation and unidirectional linear migration patterns in Myricaria laxiflora (Tamaricaceae), an endemic riparian plant in the Three Gorges Valley of the Yangtze River. AB - Myricaria laxiflora is restricted to the riverbanks of the Yangtze River valley and will be completely lost owing to the construction of the Three Gorges Dam. Genetic diversity and structure of nine natural and one ex situ populations were investigated using amplified fragment length polymorphisms (AFLPs). A moderate level of gene diversity was found in natural populations, while the ex situ population had the highest. The F statistics calculated by different approaches consistently revealed a high genetic differentiation among natural populations, contributing >45% of the total gene diversity. The Bayesian-based analysis differentiated nine independent populations in accordance with the sites sampled. Estimates of gene flow by F(ST) and coalescent-based simulation analysis indicated a restricted recurrent gene exchange among populations (Nm = 0.290 0.401), whereas genetic distance-based clustering and coalescent-based assignment analyses revealed significant genetic isolation among populations. The migration pattern in M. laxiflora is best explained by a classical metapopulation model, but with a unique unidirectional direction underlined by hydrochoric force that drove dispersal of seeds and propagules from upstream toward downstream populations. Previous efforts in preserving genomic integrity in ex situ conservation were evaluated, and the results provide valuable information to formulate conservation guidelines for successfully reintroducing M. laxiflora to the wild. PMID- 21646182 TI - Phylogeny and biogeography of the staghorn fern genus Platycerium (Polypodiaceae, Polypodiidae). AB - The genus Platycerium is one of the few pantropical epiphytic fern genera with six species in Afro-Madagascar, 8-11 Australasian species, and a single species in tropical South America. Nucleotide sequences of four chloroplast DNA markers are employed to reconstruct the phylogeny of these ferns and to explore their historical biogeography. The data set was designed to resolve conflicting hypotheses on the relationships within the genus that were based on previous phylogenetic studies exploring morphological evidence. Our results suggest a basal split of Platycerium into two well-supported clades. One clade comprises species occurring in Africa, Madagascar, and South America, whereas the second clade contains exclusively Australasian species. The latter clade is further divided into a clade corresponding to P. bifurcatum and its putative segregates and a clade of seven species occurring from Indochina throughout the Malesian region to New Guinea and Australia. The Afro-Madagascan clade includes a clade of two species found in tropical Africa and a clade of four species that includes three species endemic to Madagascar. The single neotropical species of this genus, P. andinum, is nested within the Afro-Madagascan clade but is not closely related to any extant species. PMID- 21646183 TI - Effect of flowering phenology on pollen flow distance and the consequences for spatial genetic structure within a population of Primula sieboldii (Primulaceae). AB - To evaluate the effects of flowering phenology on pollen flow distance and spatial genetic structure in a population of a bumblebee-pollinated herb, Primula sieboldii, we investigated the flowering phenology of 1712 flowers of 97 genets in a population in Nagano Prefecture, Japan, and constructed a mating model based on the observed mating pattern, which was revealed by paternity analysis using 11 microsatellite markers. The effects of flowering phenology were inferred by comparing estimated pollen flow distance and the level of heterozygosity in the next generation between two scenarios. In the first scenario, both the intergenet distance and flowering phenology influenced mating opportunity, while in the second scenario only intergenet distance influenced mating opportunity. Although the frequency distribution of pollen flow distance at the population level did not differ significantly between the two scenarios, the mean pollen flow distance of several flowers increased by more than 10 m as a result of variation in flowering phenology. Furthermore, accounting for flowering phenology predicted change in heterozygosity in the next generation from -0.04 to 0.07. The results showed that flowering phenology can affect pollen flow distance and spatial genetic structure. PMID- 21646184 TI - Self-incompatibility and floral parameters in Hypochaeris sect. Hypochaeris (Asteraceae). AB - We studied the relationships between self-incompatibility mechanisms and floral parameters in the genus Hypochaeris L. sect. Hypochaeris (consisting of H. glabra, H. radicata, H. arachnoidea, and H. salzmanniana). We assessed at intra- and interspecific levels (1) the self-incompatibility (SI) mechanism and its distribution among populations, (2) the relationship between SI and floral parameters, and (3) the relationship of SI to reproductive success. Hypochaeris salzmanniana is semi-incompatible, H. glabra is self-compatible, and H. arachnoidea and H. radicata are self-incompatible. Floral parameters differed among populations of H. salzmanniana: plants in self-compatible populations had fewer flowers per head, a smaller head diameter on the flower, and a shorter period of anthesis than self-incompatible populations. We also detected this pattern within a semi-compatible population of H. salzmanniana, and these differences were also found between species with different breeding mechanisms. Fruit to flower ratio in natural populations was generally high (>60%) in all species, regardless of breeding system. It is hypothesized that self compatibility may have arisen through loss of allelic diversity at the S locus due to bottleneck events and genetic drift. PMID- 21646185 TI - Failure of reproductive assurance in the chasmogamous flowers of Polygala lewtonii (Polygalaceae), an endangered sandhill herb. AB - Hypothetically, a species with both cleistogamous (CL) flowers and delayed selfing chasmogamous (CH) flowers should display high levels of reproductive assurance because, over time, obligate selfing by CL flowers should reduce inbreeding depression and delayed selfing in CH flowers should compensate for the absence of outcross pollen. We used pollinator-exclusion experiments to investigate reproductive assurance in the CH flowers of Polygala lewtonii, an herb with a mixed mating system. We followed CH flowers from bud-break to flower/fruit abscission to quantify fruit initiation and maturation and rates of floral development. We also evaluated the efficacy of the selfing mechanism, conducted pollinator watches to assess the likelihood of pollinator limitation, and performed regression analysis to determine the effect of flower position on fruit production. Pollinator exclusion significantly reduced fruit initiation and maturation. Investigation of floral development demonstrated that the selfing mechanism is largely dysfunctional in CH flowers, indicating the failure of reproductive assurance. Low observed rates of insect visitation appear to contradict high rates of CH fruit production in open-pollinated plants, particularly given the rarity of delayed selfing. In both treatments, flower position significantly affected fruit initiation, suggesting a role for resource limitation in both pollinator-excluded and open-pollinated flowers. PMID- 21646186 TI - Self-sterility in Ipomopsis aggregata (Polemoniaceae) is due to prezygotic ovule degeneration. AB - Based on previous studies, extreme (>99%) self-sterility in scarlet gilia (Ipomopsis aggregata) appears to be involved in late-acting ovarian self incompatibility (OSI). Here, we confirm this suggestion by comparing structural events that follow from cross- vs. self-pollinations of I. aggregata. Growth of cross- and self-pollen tubes in the style at 11 h and growth in the ovary at 24 h was equivalent. Nonetheless, by 24 h, cross-pollen effected a significantly higher percentage of both ovule penetration and fertilization. Ovules in self pollinated flowers showed pronounced changes, including an absence of embryo sac expansion and reduced starch in the integument, by 11 h post-pollination, well before pollen tube entry into the ovary. In addition, the integumentary tapetum and adjacent 1-3 cell layers exhibited abnormal cell division, pronounced deposition of thick, pectin-rich cell walls, and cellular collapse. Ovules and embryo sacs from cross-pollinated flowers rarely showed such features. Developmental changes in ovules from self-pollinated flowers eventually resulted in integument and embryo sac collapse, a process not observed in ovules of unpollinated flowers. We suggest that OSI involves long-distance signaling between self-pollen or self-pollen tubes and carpel tissue that reduces availability of receptive ovules for fertilization before pollen tubes arrive in the ovary. PMID- 21646187 TI - Female compensation through the quantity and quality of progeny in a gynodioecious plant, Geranium maculatum (Geraniaceae). AB - One of the major evolutionary trends in flowering plants is the evolution of unisexual flowers (male or female) from perfect flowers. This transition has occurred repeatedly in many taxa and has generated a wonderful array of variation in sexual expression among species. Theoretical studies have proposed a number of mechanisms to explain how this level of variation could be maintained in natural systems. One possible mechanism is the female compensation hypothesis, which predicts that female mutants require an increase in their seed fitness in order to invade a hermaphroditic system. Using Geranium maculatum, I tested this hypothesis and showed that female mothers produced more and larger seeds than hermaphroditic mothers even though they were indistinguishable in their vegetative traits and the flower production. Seeds from females were also more likely to germinate and produced seedlings with larger above- and belowground biomass. These seedlings were more likely to flower than those from hermaphrodites in at least one of the two populations studied. Combined, these results indicated that females in G. maculatum did compensate for their loss of male function by producing more and better seeds than hermaphrodites. This provides a mechanism for the maintenance of female plants in this species. PMID- 21646188 TI - A quantitative synthesis of pollen supplementation experiments highlights the contribution of resource reallocation to estimates of pollen limitation. AB - Our understanding of pollen limitation depends on a realistic view of its magnitude. Previous reviews of pollen supplementation experiments concluded that a majority of plant species suffers from pollen limitation and that its magnitude is high. Here, we perform a meta-analysis and find evidence that publication bias, experimental design, and the response variable chosen all influence the magnitude of pollen limitation. Fail-safe numbers indicate that publication bias exists for some measures of pollen limitation; significant results are more likely to be published and therefore available for review. Moreover, experiments conducted on only a fraction of a plant's flowers and reproductive episodes report ~8-fold higher effect sizes than those on all flowers produced over the entire lifetime, likely because resource reallocation among flowers and across years contributes to estimates of pollen limitation. Studies measuring percentage fruit set report higher values of pollen limitation than those measuring other response variables, such as seeds per fruit, perhaps because many plant species will not produce fruits unless adequate pollen receipt occurs to fertilize most ovules. We offer suggestions for reducing the bias introduced by methodology in pollen supplementation experiments and discuss our results in the context of optimality theory. PMID- 21646189 TI - Phylogenetic analysis of the grape family (Vitaceae) based on three chloroplast markers. AB - Seventy-nine species representing 12 genera of Vitaceae were sequenced for the trnL-F spacer, 37 of which were subsequently sequenced for the atpB-rbcL spacer and the rps16 intron. Phylogenetic analysis of the combined data provided a fairly robust phylogeny for Vitaceae. Cayratia, Tetrastigma, and Cyphostemma form a clade. Cyphostemma and Tetrastigma are each monophyletic, and Cayratia may be paraphyletic. Ampelopsis is paraphyletic with the African Rhoicissus and the South American Cissus striata nested within it. The pinnately leaved Ampelopsis form a subclade, and the simple and palmately leaved Ameplopsis constitutes another with both subclades containing Asian and American species. Species of Cissus from Asia and Central America are monophyletic, but the South American C. striata does not group with other Cissus species. The Asian endemic Nothocissus and Pterisanthes form a clade with Asian Ampelocissus, and A. javalensis from Central America is sister to this clade. Vitis is monophyletic and forms a larger clade with Ampelocissus, Pterisanthes, and Nothocissus. The eastern Asian and North American disjunct Parthenocissus forms a clade with Yua austro-orientalis, a species of a small newly recognized genus from China to eastern Himalaya. Vitaceae show complex multiple intercontinental relationships within the northern hemisphere and between northern and southern hemispheres. PMID- 21646190 TI - Phylogenetic relationships within Senna (Leguminosae, Cassiinae) based on three chloroplast DNA regions: patterns in the evolution of floral symmetry and extrafloral nectaries. AB - Senna (Leguminosae) is a large, widespread genus that includes species with enantiostylous, asymmetric flowers and species with extrafloral nectaries. Clarification of phylogenetic relationships within Senna based on parsimony analyses of three chloroplast regions (rpS16, rpL16, and matK) provides new insights on the evolution of floral symmetry and extrafloral nectaries. Our results support the monophyly of only one (Psilorhegma) of the six currently recognized sections, while Chamaefistula, Peiranisia, and Senna are paraphyletic, and monotypic Astroites and Paradictyon are nested within two of the seven major clades identified by our molecular phylogeny. Two clades (I, VII) include only species with monosymmetric flowers, while the remaining clades (II-VI) contain species with asymmetric, enantiostylous flowers, in which either the gynoecium alone or, in addition, corolla and androecium variously contribute to the asymmetry. Our results further suggest that flowers were ancestrally monosymmetric with seven fertile stamens and three adaxial staminodes, switched to asymmetry later, and reverted to monosymmetry in clade VII. Fertility of all 10 stamens is a derived state, characterizing the Psilorhegma subclade. Extrafloral nectaries evolved once and constitute a synapomorphy for clades IV VII ("EFN clade"). These nectaries may represent a key innovation in plant defense strategies that enabled Senna to undergo large-scale diversification. PMID- 21646191 TI - Untangling the phylogeny of neotropical lianas (Bignonieae, Bignoniaceae). AB - The tribe Bignonieae (Bignoniaceae) is a large and morphologically diverse clade of neotropical lianas. Despite being a conspicuous component of the neotropical flora, the systematics of the tribe has remained uncertain due to confusing patterns of morphological variation within the group. Chloroplast (ndhF) and nuclear (PepC) DNA sequences were used here to reconstruct the phylogeny of Bignonieae. Individual analyses of ndhF and PepC were highly similar to one another, yet localized differences in the placement of six species suggests some conflict between data sets. Combined analyses result in trees that are consistent with those from the individual analyses and provide greater support for the suggested relationships. This phylogeny provides important new insights into the systematics of the tribe. It identifies 21 strongly supported species groups, eight of which broadly correspond to currently recognized genera. In addition, each of these 21 species groups is supported by morphological synapomorphies. The consistency between morphological and molecular data suggests that the current phylogeny provides a solid framework for a formal revision of the generic-level classification and for addressing other aspects of the biology of Bignonieae. PMID- 21646193 TI - Errata. PMID- 21646192 TI - Molecular confirmation of the hybrid origin of Eupatorium godfreyanum (Asteraceae). AB - Analysis of nuclear ribosomal ITS sequence data was used to assess the relationships of Eupatorium godfreyanum, an agamospermous polyploid species of putative hybrid origin. A data set of ITS sequences that included representatives of all but two of the North American species of Eupatorium was compiled from a combination of previously published and newly obtained results. Assessment of the data showed that each species was relatively distinctive, although the results from parsimony analysis suggested that there was little phylogenetic structure within the data beyond a basal split between members of the dog fennel group ("Traganthes") and the remainder of the genus. Cloning was required to obtain readable ITS sequence from E. godfreyanum, and analysis of individual clones produced sequences that matched closely those of either E. rotundifolium or E. sessilifolium. The ITS sequence data thus supported the hypothesis that Eupatorium godfreyanum is of hybrid origin from a combination of E. rotundifolium and E. sessilifolium. PMID- 21646194 TI - Calceolariaceae: floral development and systematic implications. AB - The recent establishment of the new family Calceolariaceae, separated from Scrophulariaceae on the basis of molecular evidence, is complemented here by a scanning electron microscopy study of floral morphology and development of 12 species encompassing all genera (Calceolaria, Jovellana, and Stemotria [= Porodittia]). All species showed a similar pattern of organ initiation. The slightly zygomorphic, four-merous calyx is the first floral organ series initiated, with the primordia emerging consecutively in a unidirectional (dorso ventral) succession. The two entire corolla lips in Calceolaria and Jovellana arise as uniform meristematic ridges (sometimes with a central emargination, especially in Jovellana), kept apart by two lateral stamen primordia. Later the margins of the lips fuse across the backs of the young stamens, giving rise to the short corolla tube (late sympetaly). Stemotria stands out by having three stamens instead of two and a bilobed lower lip, resulting in a trimerous corolla. Similar architecture was found in teratological flowers of Calceolaria. The perianth of Calceolariaceae is shown to be derived from a tetramerous condition, not from pentamery as traditionally believed. This is in agreement with the separation of Calceolariaceae from Scrophulariaceae and with their placement in succession of Oleaceae and Tetrachondraceae in the basal Lamiales. The hitherto puzzling molecular evidence is thus supported by morphological-developmental features of the flower. PMID- 21646195 TI - Pollen ontogeny in Brasenia (Cabombaceae, Nymphaeales). AB - Brasenia is a monotypic genus sporadically distributed throughout the Americas, Asia, Australia, and Africa. It is one of eight genera that comprise the two families of Nymphaeales, or water lilies: Cabombaceae (Brasenia, Cabomba) and Nymphaeaceae (Victoria, Euryale, Nymphaea, Ondinea, Barclaya, Nuphar). Evidence from a range of studies indicates that Nymphaeales are among the most primitive angiosperms. Despite their phylogenetic utility, pollen developmental characters are not well known in Brasenia. This paper is the first to describe the complete pollen developmental sequence in Brasenia schreberi. Anthers at the microspore mother cell, tetrad, free microspore, and mature pollen grain stages were studied using combined scanning electron, transmission electron, and light microscopy. Both tetragonal and decussate tetrads have been identified in Brasenia, indicating successive microsporogenesis. The exine is tectate-columellate. The tetrad stage proceeds rapidly, and the infratectal columellae are the first exine elements to form. Development of the tectum and the foot layer is initiated later during the tetrad stage, with the tectum forming discontinuously. The endexine lamellae form during the free microspore stage, and their development varies in the apertural and non-apertural regions of the pollen wall. Degradation of the secretory tapetum also occurs during the free microspore stage. Unlike other water lilies, Brasenia is wind-pollinated, and several pollen characters appear to be correlated with this pollination syndrome. The adaptive significance of these characters, in contrast to those of the fly-pollinated genus Cabomba, has been considered. Brasenia does not produce pollenkitt nor develop tectal microchannels as does Cabomba. Instead, the discontinuity of the tectum reduces the amount of sporopollenin in the wall, which may allow for more effective wind dispersal. The importance of reassessing palynological characters in light of new ontogenetic data and the phylogenetic implications of this reevaluation are also discussed. PMID- 21646196 TI - Architecture and size relations: an essay on the apple (Malus x domestica, Rosaceae) tree. AB - The influence of tree size independent of age on some architectural features (annual shoot length, lateral branching, flowering) was investigated on 4-yr-old apple (Malus * domestica) trees either own-rooted or grafted on the dwarfing rootstock M.9, giving rise to large and small trees, respectively. Tree size significantly affected the length of the first annual shoot of bottom branches with a lesser effect on the subsequent annual shoots of the same branches and on branches situated higher in the tree canopy. The linear regression parameters, i.e., slopes and intercepts, between annual shoot length and number of growing laterals were affected by the genotype and, depending on genotype, by tree size. Flowering was generally lower, delayed, and more irregular on large trees compared to small trees, with on average similar ranking of genotypes regardless of tree size. This study provides evidence for a specific effect of tree size, as affected by the root system, on architectural development of the apple tree regardless of the genotype. From an architectural viewpoint, the dwarfing mechanism could be interpreted as a faster physiological aging essentially related to the reduction in length of the first annual shoot of bottom branches and the high flowering on this shoot. PMID- 21646197 TI - Simulated seed predation reveals a variety of germination responses of neotropical rain forest species. AB - Seed predation, an omnipresent phenomenon in tropical rain forests, is an important determinant of plant recruitment and forest regeneration. Although seed predation destroys large amounts of the seed crop of numerous tropical species, in many cases individual seed damage is only partial. The extent to which partial seed predation affects the recruitment of new individuals in the population depends on the type and magnitude of alteration of the germination behavior of the damaged seeds. We analyzed the germination dynamics of 11 tropical woody species subject to increasing levels of simulated seed predation (0-10% seed mass removal). Germination response to seed damage varied considerably among species but could be grouped into four distinct types: (1) complete inability to germinate under damage >=1%, (2) no effect on germination dynamics, (3) reduced germination with increasing damage, and (4) reduced final germination but faster germination with increasing damage. We conclude that partial seed predation is often nonlethal and argue that different responses to predation may represent different proximal mechanisms for coping with partial damage, with potential to shape, in the long run, morphological and physiological adaptations in tropical, large-seeded species. PMID- 21646198 TI - Ancestral chloroplast polymorphism and historical secondary contact in a broad hybrid zone of Aesculus (Sapindaceae). AB - Knowledge regarding the origin and maintenance of hybrid zones is critical for understanding the evolutionary outcomes of natural hybridization. To evaluate the contribution of historical contact vs. long-distance gene flow in the formation of a broad hybrid zone in central and northern Georgia that involves Aesculus pavia, A. sylvatica, and A. flava, three cpDNA regions (matK, trnD-trnT, and trnH trnK) were analyzed. The maternal inheritance of cpDNA in Aesculus was confirmed via sequencing of matK from progeny of controlled crosses. Restriction site analyses identified 21 unique haplotypes among 248 individuals representing 29 populations from parental species and hybrids. Haplotypes were sequenced for all cpDNA regions. Restriction site and sequence data were subjected to phylogeographic and population genetic analyses. Considerable cpDNA variation was detected in the hybrid zone, as well as ancestral cpDNA polymorphism; furthermore, the distribution of haplotypes indicates limited interpopulation gene flow via seeds. The genealogy and structure of genetic variation further support the historical presence of A. pavia in the Piedmont, although they are at present locally extinct. In conjunction with previous allozyme studies, the cpDNA data suggest that the hybrid zone originated through historical local gene flow, yet is maintained by periodic long-distance pollen dispersal. PMID- 21646199 TI - The role of fungal pathogens in flower size and seed mass variation in three species of Hydrophyllum (Hydrophyllaceae). AB - Identifying ecological factors that affect seed number and seed size is key to understanding the persistence of large seed mass variation in some plant species. Pathogens may increase seed mass variation by increasing resource demand over the growing season such that late fruits experience higher resource competition than early fruits. We tested whether Fusarium sp. and Rhizoctonia sp., soil fungi that cause wilt, contributed to seasonal decline in flower size, seed number, or seed mass in Hydrophyllum appendiculatum and H. canadense. A third species not infected by these soil fungi, H. virginianum, was studied to determine how seasonal decline in floral traits and seed mass variation varies within this genus. Flower size declined seasonally for all species, but was greatest for H. appendiculatum, a monocarpic biennial with indeterminate inflorescences. Seed number decreased between first and last inflorescences in H. appendiculatum, but not in H. canadense or H. virginianum, perennials with determinate inflorescences. Seed mass varied most in H. appendiculatum and H. canadense (4-20 fold in 50% of individuals) and least in H. virginianum (4-8-fold in >30% of individuals). Fungal infection increased seed mass variation among diseased plants in H. canadense and H. appendiculatum. However, within plants fungal infection only increased seasonal decline in flower size, seed number, and seed mass in H. appendiculatum when flowers received supplemental pollination. PMID- 21646200 TI - Molecular phylogeny of the Caryophyllaceae (Caryophyllales) inferred from chloroplast matK and nuclear rDNA ITS sequences. AB - Caryophyllaceae is a principally holarctic family including around 2200 species often classified into the three subfamilies Alsinoideae, Caryophylloideae, and Paronychioideae. Complex and possibly homoplasious morphological characters within the family make taxa difficult to delimit and diagnose. To explore part of the morphological evolution within the family, we investigated the phylogeny of the Caryophyllaceae by means of analyzing plastid and nuclear sequence data with parsimony and Bayesian methods. We describe a mode of tracing a stable phylogenetic signal in ITS sequences, and a significant common signal is shared with the plastid data. Parsimony and Bayesian analyses yield some differences in tree resolution. None of the subfamilies appear monophyletic, but the monophyly of the Caryophylloideae is not contradicted. Alsinoideae are paraphyletic, with Arenaria subg. Eremogone and Minuartia subg. Spergella more closely related to the Caryophylloideae. There is strong support for the inclusion of Spergula Spergularia in an Alsinoideae-Caryophylloideae clade. Putative synapomorphies for these groupings are twice as many stamens as number of sepals and a caryophyllad type of embryogeny. Paronychioideae form a basal grade, where tribe Corrigioleae are sister to the rest of the family. Free styles and capsules with simple teeth are possibly plesiomorphic for the family. PMID- 21646201 TI - Polyploid and hybrid evolution in roses east of the Rocky Mountains. AB - This study investigates the impact of hybridization and polyploidy in the evolution of eastern North American roses. We explore these processes in the Rosa carolina complex (section Cinnamomeae), which consists of five diploid and three tetraploid species. To clarify the status and origins of polyploids, a haplotype network (statistical parsimony) of the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) nuclear gene was estimated for polyploids of the complex and for diploids of section Cinnamomeae in North America. A genealogical approach helped to decipher the evolutionary history of polyploids from noise created by hybridization, incomplete lineage sorting, and allelic segregation. At the diploid level, species west of the Rocky Mountains are distinct from eastern species. In the east, two groups of diploids were found: one consists of R. blanda and R. woodsii and the other of R. foliolosa, R. nitida, and R. palustris. Only eastern diploids are involved in the origins of the polyploids. Rosa arkansana is derived from the blanda-woodsii group, R. virginiana originated from the foliolosa-nitida-palustris group, and R. carolina is derived from a hybrid between the two diploid groups. The distinct origins of these polyploid taxa support the hypothesis that the three polyploids are separate species. PMID- 21646202 TI - The revision of species-rich genera: a phylogenetic framework for the strategic revision of Pilea (Urticaceae) based on cpDNA, nrDNA, and morphology. AB - The revision of species-rich genera underpins research and supports the sustainable use and monitoring of biological diversity. One fifth to one quarter of the diversity of all seed plant species occurs in such genera, but difficulties with the revision of species-rich genera has resulted in many of them being ignored since the late 1800s. Pilea, with 600-715 species is in need of revision. The only realistic approach is in manageable subunits, which requires confirmation of monophyly and identification of monophyletic subdivisions. Parsimony analyses of trnL-F, ITS, and morphology data were used to test the monophyly of, and explore intrageneric relationships within, Pilea. Analysis of trnL-F data confirms and recovers two morphologically diagnosable monophyletic clades that include all of the taxa within Pilea. Overlaying geographic distribution on a most parsimonious tree indicates a strong association between geography and phylogenetic relatedness. It is suggested that a strategic revision within the framework of morphologically and geographically diagnosable units might enable the revision of the group using an iterative approach. Analysis of the outgroup taxa supports the inclusion of Poikilospermum within the Urticaceae and suggests that the Urticaceae tribes could be placed into two clades that are supported by floral morphology. PMID- 21646203 TI - Chloroplast DNA variation and geographical structure of the Aristolochia kaempferi group (Aristolochiaceae). AB - The present study documents cpDNA variation in the Aristolochia kaempferi group (Aristolochiaceae), which consists of one Chinese and all Japanese and Taiwanese species of the subgenus Siphisia. In a phylogenetic analysis based on the nucleotide sequences of the matK gene, and the atpB-rbcL and trnS-trnG intergenic spacer regions, 38 haplotypes were recognized in the A. kaempferi group and as many as 24 within A. kaempferi. This is the most haplotypes reported for a single species to date. Although six highly significant major clades were identified in the phylogenetic analysis, they were not congruent with previous classifications. This might be attributed to the specific speciation process, such as convergent evolution, incomplete lineage sorting, and/or reticulate evolution. The six major clades had a clear geographical distribution pattern and were significantly associated with geographical distribution of haplotypes in a nested clade analysis and AMOVA. The results allow us to deduce a scenario in which multiple contractions and expansions of the geographical ranges brought about by Quaternary climatic oscillations affected the patterns of genetic diversity. The present geographic patterns of haplotype distribution within the A. kaempferi group can be explained by the last postglacial range expansion from different refugia, and the boundaries may be suture zones. PMID- 21646204 TI - Rare plants are common where you find them. AB - Broad patterns in distribution and abundance can elucidate processes of evolution. A positive association between local abundance and the size of the geographic range has been demonstrated for closely related species across many taxa. This pattern is usually explained by assuming that species with smaller ranges are ecologically inferior (e.g., poor competitors or dispersers). Many areas of high endemism support local species that have evolved recently. The distribution of these neoendemics may reflect historical processes not accounted for by ecological, equilibrium hypotheses. We asked whether such traditional macroecological hypotheses also applied to the local abundance of seven narrowly endemic species and ecologically similar widespread congeners in the northern Rocky Mountains. For each of the 14 species, we estimated abundance of five randomly chosen populations by counting plants in 10 randomly located plots. The association between range size and local abundance was not positive. Instead, all seven narrow endemics were more abundant than their widespread congeners. Ecological specialization or differences in dispersal ability are not likely explanations for our results. We believe the local abundance of narrowly endemic species may be a sign of recent speciation. Most or all of our narrowly distributed species have probably not yet had time to spread to their full potential. Furthermore, theory predicts that speciation is more likely to occur in locally abundant populations. Our results suggest that strictly ecological mechanisms cannot explain abundance and distribution in regions with high neoendemism. PMID- 21646205 TI - Molecular phylogeography and hybridization in members of the circumpolar Potentilla sect. Niveae (Rosaceae). AB - Glacial events and the formation of ice-free areas serving as refugia for plants and animals are important in shaping present patterns of genetic diversity in arctic areas. Beringia, situated in northeastern Russia and Alaska, has been pointed out as a major refugium. This study focuses on the historical biogeography of the circumpolar taxon Potentilla sect. Niveae. The taxonomy of the group is complex, most likely highly influenced by hybridization and apomixis. cpDNA microsatellites together with AFLP fragments were used to map the genetic variability in the section, from Beringia across the Canadian Arctic to Greenland. The data support the hypothesis that Beringia, as well as parts of adjacent arctic Canada, served as refugia during the Wisconsinan glaciation, and there is some evidence for a northern and a southern migration route out of Beringia. The hair type groups within sect. Niveae are more or less genetically distinct, and hybridization, especially with sect. Multifida, takes place. Haplotype diversity as well as frequency is at its maximum close to the Last Glacial Maximum ice cap edge. This pattern can be explained by merging of previously isolated refugia, by repeated extinction/colonization events close to the ice edge, and by hybridization among sympatric taxonomical lineages. PMID- 21646206 TI - Intraspecific nuclear ribosomal DNA divergence and reticulation in sexual diploid Erigeron strigosus (Asteraceae). AB - Apomictic complexes in flowering plants often harbor multiple sexual taxa, which must be studied to interpret accurately the origin and diversity of apomictic derivatives. The daisy fleabane group in North America (Erigeron sect. Phalacroloma) includes the widespread polyploid apomictic taxa E. annuus, E. strigosus, and E. tenuis, as well as recently discovered sexual diploid populations in the southeastern United States (E. strigosus var. calcicola, E. strigosus var. dolomiticola, sexual E. strigosus var. strigosus). Phylogenetic analysis for 35 sexual populations was conducted using sequences from the internal and external transcribed spacers (ITS and ETS) of ribosomal DNA. Results indicate that the three groups of sexual plants form separate monophyletic clades and that edaphic specialization is ancestral in the group. Sequence analysis for sexual E. strigosus var. strigosus was confounded by the fact that 13 of 31 samples (42%) possessed nucleotide polymorphisms at 1.0-1.8% of sites. However, analysis of the genomic sequence for plants with low polymorphism levels in conjunction with analysis of cloned PCR products for plants with high levels of intra-individual polymorphism indicated that three combinations of haplotypes (N I + N II, two plants; N I + N III, five plants; N II + N III, six plants) accounted for the majority of sequence polymorphisms encountered. These data are consistent with historical diversification and subsequent reticulate evolution at the diploid level in sexual E. strigosus var. strigosus. PMID- 21646207 TI - Species divergence and relationships in Stephanomeria (Compositae): PgiC phylogeny compared to prior biosystematic studies. AB - We present a maximum likelihood tree of 41 PgiC sequences for the monophyletic Stephanomeria, with 10 perennial and six annual species, widely distributed in western North America and exemplary of different speciation processes. The phylogenetic analysis represents the first use of PgiC sequences for Compositae. The annual species were originally delimited by biosystematic studies that provided evidence of their reproductive compatibility and chromosome structural homology. The perennial species are highly distinctive in morphology and have not been examined similarly. The PgiC tree provides more resolution than our previous ITS/ETS tree and reflects both past and ongoing hybridization and/or incomplete lineage sorting. Two major PgiC clades were resolved in Stephanomeria. One clade contains the genes from the annual species plus the perennial, insular endemic S. guadalupensis, which appears closely related to a monophyletic S. virgata. Stephanomeria exigua is not monophyletic. The second clade includes the genes from all other sampled perennial species and a monophyletic subclade of four genes from two annual species. The results are compared to previous studies, also using PgiC, of Clarkia (Onagraceae). Both molecular systematic and biosystematic approaches are essential to discern the very different courses of evolution in these two, well-studied genera of western North America. PMID- 21646208 TI - Loss of extrafloral nectary on an oceanic island plant and its consequences for herbivory. AB - Two Hibiscus (Malvaceae) species coexist on the oceanic Bonin (Ogasawara) Islands: Hibiscus glaber (an endemic species) and H. tiliaceus (the ancestral non endemic species). Hibiscus tiliaceus produces extrafloral nectar from the sepals, while H. glaber does not. To clarify the effects of extrafloral nectar loss on Hibiscus-insect relationships, we examined herbivory and insect communities on flower buds of H. glaber and H. tiliaceus. Larvae of the endemic moth Rehimena variegata (Lepidoptera: Pyralidae) attacked 20% of the flower buds on H. glaber, while less than 0.2% of buds on H. tiliaceus were attacked. Introduced species of ants frequently visited the flower buds of H. tiliaceus to collect extrafloral nectar from the sepal, while they rarely visited those of H. glaber. Therefore, extrafloral nectar on H. tiliaceus sepals may function as a facultative defense against flower bud herbivory. The loss of extrafloral nectaries of H. glaber sepals may be related to the original paucity of native herbivores and ants on the Bonin Islands. PMID- 21646209 TI - The structure of xylem vessels in grapevine (Vitaceae) and a possible passive mechanism for the systemic spread of bacterial disease. AB - Xylem-dwelling pathogens become systemic, suggesting that microorganisms move efficiently in the xylem. To better understand xylem pathways and how bacteria move within the xylem, vessel connectivity between stems and leaves of Vitis vinifera cv. Chardonnay and Muscadinia rotundifolia cv. Cowart was studied. Three methods were used: (1) the light-producing bacterium, Yersinia enterocolitica, (Ye) strain GY5232 was loaded into petioles and followed using X-ray film, (2) fluorescent beads were loaded and followed by microscopy, and (3) low-pressure air was pumped into leaves and extruded bubbles from cuts in submerged leaves were followed. Bacteria, beads, and air moved through long and branched xylem vessels from the petiole into the veins in leaves of both varieties. From the stem, bacteria and air traveled into primary and secondary veins of leaves one, two, and three nodes above the loading point of the bacteria or air. Particles and air could move unimpeded through single xylem vessels or multiple vessels (conduits) connected possibly through broken pit membranes from within the stem axis into leaf blades. Bacteria were also able to move long distances within minutes from stem to leaf passively without having to cross pit membranes. Such complex, open xylem conduits have not been well documented before; these findings will help elucidate mechanisms involved in the systemic spread of pathogens. PMID- 21646210 TI - Aborted fruits of Opuntia microdasys (Cactaceae): insurance against reproductive failure. AB - New individuals in clonal populations arise through the recruitment of sexual or clonal offspring. The predominance of one type of regeneration over the other has been correlated with different selective environmental pressures. We compared the reproductive mode (sexual through seeds and vegetative through plantlets or detached cladodes) of Opuntia microdasys from three desert habitats of the Chihuahuan Desert: bajada (BH), hill-piedmont (HPH), and an interdune (IDH). Successful establishment and growth of plantlets were determined in two experiments: (1) the effect of light (three levels of photosynthetically active radiation [PAR]: full, low, and medium) and two levels of watering and (2) maternal effects and provenance of plantlets. Adult plant densities did not differ among habitats (639 individuals/ha), but the number of offspring and fruit production increased significantly at BH. Plantlets (94.3%) dominated the form of recruitment for all habitats, followed by cladodes (3.1%) and seedlings (2.6%). A higher proportion of plantlets established in the low and medium PAR treatments (76%) in comparison to full exposure (39%). Maternal factors affected survival and growth, but plantlet provenance did not. The high fruit abortion rate resulting from environmental and maternal effects provided suitable conditions for establishment of plantlets. PMID- 21646211 TI - A new herbarium-based method for reconstructing the phenology of plant species across large areas. AB - Phenological data have recently emerged as particularly effective tools for studying the impact of climate change on plants, but long phenological records are rare. The lack of phenological observations can nevertheless be filled by herbarium specimens as long as some correction procedures are applied to take into account the different climatic conditions associated with sampling locations. In this study, we propose a new herbarium-based method for reconstructing the flowering dates of plant species that have been collected across large areas. Coltsfoot (Tussilago farfara L.) specimens from southern Quebec were used to test the method. Flowering dates for coltsfoot herbarium specimens were adjusted according to the date of disappearance of snow cover in the region where they were collected and compared using a reference point (the date of earliest snowmelt). In southern Quebec, coltsfoot blooms earlier at present (15-31 d) than during the first part of the 20th century. This phenomenon is likely associated with the climate warming trends recorded in this region in the last century, especially during the last three decades when the month of April became warmer, thereby favoring very early-flowering cases. The earlier flowering of coltsfoot is, however, only noticeable in large urban areas (Montreal, Quebec City), suggesting a strong urban heat island effect on the flowering of this plant. Herbarium specimens are useful phenological indicators; however, the databases should be carefully examined prior to analysis to detect biases or trends associated with sampling locations. PMID- 21646212 TI - Variability in leaf optical properties of Mesoamerican trees and the potential for species classification. AB - Leaf traits and physiological performance govern the amount of light reflected from leaves at visible and infrared wavebands. Information on leaf optical properties of tropical trees is scarce. Here, we examine leaf reflectance of Mesoamerican trees for three applications: (1) to compare the magnitude of within and between-species variability in leaf reflectance, (2) to determine the potential for species identification based on leaf reflectance, and (3) to test the strength of relationships between leaf traits (chlorophyll content, mesophyll attributes, thickness) and leaf spectral reflectance. Within species, shape and amplitude differences between spectra were compared within single leaves, between leaves of a single tree, and between trees. We also investigated the variation in a species' leaf reflectance across sites and seasons. Using forward feature selection and pattern recognition tools, species classification within a single site and season was successful, while classification between sites or seasons was not. The implications of variability in leaf spectral reflectance were considered in light of potential tree crown classifications from remote airborne or satellite-borne sensors. Species classification is an emerging field with broad applications to tropical biologists and ecologists, including tree demographic studies and habitat diversity assessments. PMID- 21646213 TI - Plasticity of inflorescence traits in Lobelia siphilitica (Lobeliaceae) in response to soil water availability. AB - Many workers have demonstrated a genetic basis for variation in inflorescence traits, but this variation can also have an environmental component. Because flowering can incur significant water costs, I estimated plasticity of inflorescence traits of three populations of Lobelia siphilitica in response to drought. I manipulated soil water availability in the greenhouse and measured seven inflorescence traits. Under drought conditions, plants from one population flowered later and produced fewer flowers with shorter corollas and narrower landing pads. In contrast, the height of the flowering stalk decreased in response to drought in all three populations. Consequently, pollinator-mediated natural selection on these plastic traits may depend on soil water availability. Plastic responses differed between genotypes only for the height of the flowering stalk and the length of the corolla tube and only in one or two populations. This suggests that genotype * environment interactions would not limit the evolution of inflorescence traits in L. siphilitica. The strength and sign of phenotypic correlations among inflorescence traits did not respond plastically to drought, suggesting that indirect selection on inflorescence traits of L. siphilitica will not vary strongly with water availability. My results suggest that plasticity of inflorescence traits may influence their evolution, but the effects are population- and trait-specific. PMID- 21646214 TI - A modular approach to biomass allocation in an invasive annual (Microstegium vimineum; Poaceae). AB - The invasive grass Microstegium vimineum grows in low light beneath the canopy of eastern forests in North America by reiteration of modules (phytomers) along a tiller. Basal phytomers are vegetative; terminal phytomers produce a raceme of chasmogamous (CH) spikelets plus an axillary raceme of cleistogamous (CL) spikelets. Additional subterminal phytomers with CL racemes mature basipetally. Allocation to culms, leaves, and CH and CL within phytomers was examined in relation to light conditions for a population in New Jersey, USA. Plants were reared in a greenhouse from seed families of parents in deep shade (2-8% full sun) or sunny, edge habitats. Primary tillers were subdivided into phytomers, dried, and weighed. Tillers from field habitats were similarly treated. For vegetative and subterminal phytomers, allocation to leaves and CH was greatest for the shady habitat. CL allocation decreased from terminal to reproductively immature subterminal phytomers. CH and CL mass was positively correlated with leaf mass, suggesting that reproduction is determined by available photosynthate. CH mass showed a genetic correlation with leaf mass. Developmental plasticity in modular allocation allows Microstegium to maximize fitness when conditions are favorable (e.g., high light along forest edges) by continual maturation of CL caryopses on axillary racemes. PMID- 21646215 TI - Evolutionary relationships of Mycaureola dilseae (Agaricales), a basidiomycete pathogen of a subtidal rhodophyte. AB - Mushroom-forming fungi (homobasidiomycetes) are major examples of morphological and ecological diversification in terrestrial habitats. Homobasidiomycetes includes only nine described species that are known from marine environments. Morphological traits that have concealed the ancestry of these fungi include reduced fruiting bodies with hairy surfaces and extremely modified spores, both of which may function as floating devices to aid successful dispersal and adhesion to various substrates such as driftwood. Our previous results suggested that all marine forms as yet investigated are placed in the Nia clade (euagarics) and that they have primarily evolved from cypelloid forms (minute, cup-shaped, terrestrial saprotrophs) via transitions through mangroves to fully marine habitats. We show here that Mycaureola dilseae, which parasitizes the red alga Dilsea carnosa, is a second independent lineage of marine fungi in the euagarics clade that is not related to cyphelloid forms. Phylogenetic reconstructions were based on two data sets: a partial four-region rDNA data set (nuc-ssu, nuc-lsu, mt ssu, and mt-lsu) with inclusive sampling of 249 taxa and a densely sampled ITS data set including 32 taxa, which formed a clade with Mycaureola in the four region rDNA analyses. Inferences using constrained and unconstrained six parameter weighted parsimony, Bayesian Markov chain Monte Carlo methods, and maximum likelihood approaches place M. dilseae in the morphologically diverse /physalacriaceae clade next to Gloiocephala spp., a group of highly reduced stipitate-pileate saprotrophs. PMID- 21646216 TI - Beardia vancouverensis gen. et sp. nov. (Juglandaceae): permineralized fruits from the Eocene of British Columbia. AB - Large numbers of permineralized juglandaceous fruits were identified in calcareous nodules from the Eocene Appian Way locality on Vancouver Island, British Columbia, Canada. The fruits, small dorsiventrally flattened nutlets, 4.5 7.0 mm long and 5.5-9.0 * 3-5 mm in diameter, were studied using cellulose acetate peels. They are wingless, ribbed, and have a lobed epicarp that surrounds the nutlet. Cells of the inner epicarp are thin-walled and traversed by a system of branching vascular strands. The stony nutlet wall is composed of fibers, with an outer layer of distinctive idioblasts. The fruits have a symmetry like that in Juglandaceae, subfamily Juglandoideae, tribe Platycaryeae, while the fibrous nut walls are like those of subfamily Engelhardioideae. This unique combination of characters indicates that these fruits represent a new genus and species of Juglandaceae: Beardia vancouverensis gen. et sp. nov. The excellent preservation of the Appian Way specimens has allowed a unique view of the internal fruit anatomy and external morphology. As the only wingless, flattened nuts known in the family, they further extend the range of morphological variation in fruits in the family. These fossils further support the hypothesis that North America was an important center of generic diversity for Juglandaceae during the early Tertiary. PMID- 21646217 TI - Development of a comprehensive detection method for medicinal and toxic plant species. AB - Pharmacologically active ingredients in plants can cause significant morbidity through their increasingly common use in herbal alternative medicines and dietary supplements. Monitoring consumer products for the presence of toxic plants is encumbered by the lack of rapid and specific assays. To create a sensitive, reliable, fast, and broad-spectrum assay for medicinal or toxic plant species, we tested multiplexed ligation-dependent probe amplification (MLPA), which requires partial genomic DNA sequences from species of plants that are not well represented in currently available genetic databases. Genomic DNA was obtained from 21 species of medicinal and/or toxic plants. The PCR products were amplified from these plants and cloned for sequencing. The MLPA method was successful with DNA samples from many different species. The use of a microarray to facilitate screening of potentially thousands of plants in a single assay also was successful. The combination of the specificity of the MLPA assay with the broad scale capabilities of microarray technology should make this an especially useful tool in screening in foods and commercial herbal preparations to identify the plant compounds actually present. Other applications could potentially extend to the identification of any plant species in samples for academic botanical studies and for biodefense and forensics applications. PMID- 21646218 TI - Extreme intraplant variation in nectar sugar composition in an insect-pollinated perennial herb. AB - Variation in nectar chemistry among plants, flowers, or individual nectaries of a given species has been only rarely explored, yet it is an essential aspect to our understanding of how pollinator-mediated selection might act on nectar traits. This paper describes variation in nectar sugar composition in a population of the perennial herb Helleborus foetidus (Ranunculaceae) and dissects it into components due to variation among plants, flowers of the same plant, and nectaries of the same flower. The proportions of sucrose, glucose, and fructose in single-nectary nectar samples collected at two times in the flowering season were determined using high performance liquid chromatography (HPLC). Sugar composition varied extensively among nectaries, and nearly all combinations of individual sugars were recorded. Population-wide variance was mainly accounted for by variation among flowers of the same plant (56% of total), nectaries of the same flower (30%), and only minimally by differences among plants (14%). In absolute terms, intraplant variation was similar to or greater than that ordinarily reported in interspecific comparisons. Results suggest that the prevailing notion of intraspecific constancy in nectar sugar composition may be unwarranted for some species and that more elaborate nectar sampling designs are required to detect and appropriately account for extensive within-plant variance. Within-plant variation in nectar sugar composition will limit the ability of pollinators to exert selection on nectar chemistry in H. foetidus and may be advantageous to plants by reducing the number of flowers visited per foraging bout by variance-sensitive, risk-averse pollinators. PMID- 21646219 TI - Genetic variation and reproductive system among North American species of Nuttallanthus (Plantaginaceae). AB - We examined the effect of reproductive and life history strategies on the amount and partitioning of genetic variation in three annual species of Nuttallanthus. The North American species N. canadensis, N. floridanus, and N. texanus have regional to widespread ranges that overlap in the southeastern USA, are characterized by homogeneous populations and high fecundity, and possess showy, fragrant flowers seemingly adapted for insect pollination and outbreeding. Field and greenhouse studies on plants from 25 populations indicated that reproductive strategies were similar among species and showed predominant self-fertilization via cleistogamy and self-pollination prior to anthesis in chasmogamous flowers. Species were reproductively isolated and demonstrated complete cross incompatibility after experimental crosses and no evidence for hybridization in mixed populations. Genetic variation was assessed using starch gel electrophoresis to resolve 15 isozyme loci in 50 populations. Conspecific genetic identity (I) values were high (0.819-0.936), but interspecific comparisons indicated many qualitative allelic differences and correspondingly low I values (0.516-0.623). Low levels of polymorphism and observed heterozygosity within populations and the disproportionate amount of gene diversity distributed among populations were concordant with reproductive data. The pattern of genetic differentiation was most similar to that observed in species with a predominantly inbreeding mating system. PMID- 21646220 TI - Variation in the self-incompatibility response within and among populations of the tropical shrub Witheringia solanacea (Solanaceae). AB - Breakdown of genetically enforced self-incompatibility (SI), an extremely common and important evolutionary transition in plants, has conventionally been conceived as a qualitative rather than a quantitative change. We evaluated qualitative and quantitative variation in SI for four populations of Witheringia solanacea in Costa Rica, examining growth of self-pollen tubes in pollinations of buds and mature flowers. We also measured levels of RNase production in styles to determine whether enzyme production was correlated with differences in self rejection. The two small populations contained both self-compatible (SC) individuals and obligate outcrossers (female or SI). Plants in the two large populations were uniformly SI as revealed by pollen tube growth, although several of these individuals sporadically set seed autogamously. Stylar RNase activity did not differ significantly between bud and mature flowers, but self-pollen tube growth did differ, suggesting that a gene product in addition to S-RNase is responsible for developmental onset of SI. Population-level differences in RNase activity were consistent with differences in the strength of the rejection response in bud pollinations, suggesting that a threshold level of S-RNase, in combination with other factors, is necessary for SI. Our results support a growing body of evidence that not only qualitative variation in SI, but also quantitative variation may be functionally significant. PMID- 21646221 TI - A multi-year study of factors affecting fruit production in Aristolochia paucinervis (Aristolochiaceae). AB - Pollen limitation, resource limitation, fruit abortion, and predation have all been proposed as factors explaining low fruit set in hermaphroditic plants. We conducted a 5-year study combining field observations and pollination experiments to determine the causes of the low fruit set in Aristolochia paucinervis, a Mediterranean species with a specialized pollination system in two populations in SW Spain. Fruit initiation was markedly low, and between 28.6 and 75.0% of the flowering stems did not initiate any fruit. In most flowers, the number of germinated pollen grains was less than the number of ovules, and supplemental pollination significantly increased fruiting, indicating deficient pollination. In A. paucinervis, autonomous self-pollination seems to be a decisive factor in fruit production because the number of germinated pollen and the fruit set from flowers bagged before anthesis were similar to those in free-pollinated flowers. Only in 2005 did flowers that were successfully pollinated outnumber ripened fruits, suggesting that other factors limit fruiting. We found a significant positive correlation between tuber mass and fruit set. Deficient pollination and lack of resources could explain the low fruit set, but the relative consequences seem to vary spatially and temporarily. PMID- 21646222 TI - Brassicaceae phylogeny and trichome evolution. AB - To estimate the evolutionary history of the mustard family (Brassicaceae or Cruciferae), we sampled 113 species, representing 101 of the roughly 350 genera and 17 of the 19 tribes of the family, for the chloroplast gene ndhF. The included accessions increase the number of genera sampled over previous phylogenetic studies by four-fold. Using parsimony, likelihood, and Bayesian methods, we reconstructed the phylogeny of the gene and used the Shimodaira Hasegawa test (S-H test) to compare the phylogenetic results with the most recent tribal classification for the family. The resultant phylogeny allowed a critical assessment of variations in fruit morphology and seed anatomy, upon which the current classification is based. We also used the S-H test to examine the utility of trichome branching patterns for describing monophyletic groups in the ndhF phylogeny. Our phylogenetic results indicate that 97 of 114 ingroup accessions fall into one of 21 strongly supported clades. Some of these clades can themselves be grouped into strongly to moderately supported monophyletic groups. One of these lineages is a novel grouping overlooked in previous phylogenetic studies. Results comparing 30 different scenarios of evolution by the S-H test indicate that five of 12 tribes represented by two or more genera in the study are clearly polyphyletic, although a few tribes are not sampled well enough to establish para- or polyphyly. In addition, branched trichomes likely evolved independently several times in the Brassicaceae, although malpighiaceous and stellate trichomes may each have a single origin. PMID- 21646223 TI - Patterns of anomalous floral development in the Asian Passiflora (subgenus Decaloba: supersection Disemma). AB - Approximately 22 species of Passiflora are native to the Old World. All of these species are placed in subgenus Decaloba, supersection Disemma. Within Disemma, three species vary in stamen and carpel number (<= eight stamens and five carpels). The mode of development was determined for two of the anomalous species, P. moluccana var. glaberrima and P. siamica. Ontogenetic patterns were compared to normal development in P. perakensis and P. holosericea. Passiflora siamica develops additional stamens through dedoublement of a single widened stamen primordium, while P. moluccana var. glaberrima exhibits congenital dedoublement where stamens emerge already doubled. Phylogenetic analysis using ITS and the trnL-F intron and spacer resolve the anomalous species as monophyletic and sister to P. perakensis. This signifies a single loss of genetic regulation in stamen and carpel number within Disemma. Floral whorls were examined across the Passifloraceae, Malesherbiaceae, Turneraceae, and Flacourtiaceae s.l.. Similar doubling in these families suggests that this Eurosid lineage may have a genetic propensity for variability in floral whorl number. PMID- 21646224 TI - Phylogenetics of tribe Phyllantheae (Phyllanthaceae; Euphorbiaceae sensu lato) based on nrITS and plastid matK DNA sequence data. AB - Phylogenetic relationships within tribe Phyllantheae, the largest tribe of the family Phyllanthaceae, were examined with special emphasis on the large genus Phyllanthus. Nuclear ribosomal ITS and plastid matK DNA sequence data for 95 species of tribe Phyllantheae, including representatives of all subgenera of Phyllanthus (except Cyclanthera) and several hitherto unplaced infrageneric groups, were analyzed. Results for ITS and matK are generally concordant, although some species are placed differently in the plastid and ITS trees, indicating that hybridization/paralogy is involved. Results confirm paraphyly of Phyllanthus in its traditional circumscription with embedded Breynia, Glochidion, Reverchonia, and Sauropus. We favor the inclusion of the embedded taxa in Phyllanthus over further generic segregation. Monophyletic Phyllanthus comprises an estimated 1269 species, making it one of the "giant" genera. Phyllanthus maderaspatensis is sister to all other species of Phyllanthus, and the genus appears to be of paleotropical origin. Subgenera Isocladus, Kirganelia, and Phyllanthus are polyphyletic, whereas other subgenera appear to be monophyletic. Monotypic Reverchonia is sister to P. abnormis, arborescent section Emblica to herbaceous Urinaria, free-floating aquatic P. fluitans to the weed P. caroliniensis, and the phyllocladous section Choretropsis to the delicate leafy P. claussenii. The unique branching architecture known as "phyllanthoid branching" found in most Phyllanthus taxa has been lost (and/or has been derived) repeatedly. Taxonomic divisions within Phyllantheae based on similar pollen morphology are confirmed, and related taxa share similar distributions. We recommend recognition of six clades at generic level: Flueggea s.l. (including Richeriella), Lingelsheimia, Margaritaria, Phyllanthus s.l. (including Breynia, Glochidion, Reverchonia, and Sauropus), P. diandrus, and Savia section Heterosavia. PMID- 21646225 TI - Allozyme diversity within and divergence among species ofTolpis(Asteraceae Lactuceae) in the Canary Islands: systematic, evolutionary, and biogeographical implications. AB - Plants endemic to oceanic islands represent some of the most unusual and rare taxa in the world. Enzyme electrophoresis was used to assess genetic diversity within and divergence among all endemic species of a small genus of plants on the Canary Islands. Our results show that the genus Tolpis is similar to many other island groups in having generally low allozyme divergence among species, with the highest divergence found among four groups of endemics. The two rare and highly localized species T. glabrescens and T. crassiuscula are each divergent from all other species in the Canaries. Tolpis coronopifolia is also divergent at allozyme loci; this is the only endemic species that is a self-compatible annual (or weak biennial). A large, morphologically variable species complex consisting of T. laciniata and T. lagopoda together with several named and unnamed morphological variants shows low allozyme divergence among its elements. The evolution of polyploidy from diploid ancestors in situ in oceanic archipelagos is uncommon, but the tetraploid T. glabrescens is an exception. Allozyme data do not implicate any extant diploid Tolpis species as parents of the polyploid. It is possible that T. glabrescens originated early in the evolution of Tolpis in the Canary Islands and that its parents are now extinct. The nonendemic T. barbata shows no greater divergence from the Canary Island endemics than some endemics exhibit among themselves. Both changes in allele frequencies and unique alleles are responsible for genetic divergence among species of Tolpis. PMID- 21646229 TI - Driving restrictions after implantable cardioverter defibrillator implantation: an evidence-based approach. AB - AIMS: Little evidence is available regarding restrictions from driving following implantable cardioverter defibrillator (ICD) implantation or following first appropriate or inappropriate shock. The purpose of the current analysis was to provide evidence for driving restrictions based on real-world incidences of shocks (appropriate and inappropriate). METHODS AND RESULTS: A total of 2786 primary and secondary prevention ICD patients were included. The occurrence of shocks was noted during a median follow-up of 996 days (inter-quartile range, 428 1833 days). With the risk of harm (RH) formula, using the incidence of sudden cardiac incapacitation, the annual RH to others posed by a driver with an ICD was calculated. Based on Canadian data, the annual RH to others of 5 in 100 000 (0.005%) was used as a cut-off value. In both primary and secondary prevention ICD patients with private driving habits, no restrictions to drive directly following implantation, or an inappropriate shock are warranted. However, following an appropriate shock, these patients are at an increased risk to cause harm to other road users and therefore should be restricted to drive for a period of 2 and 4 months, respectively. In addition, all ICD patients with professional driving habits have a substantial elevated risk to cause harm to other road users during the complete follow-up after both implantation and shock and should therefore be restricted to drive permanently. CONCLUSION: The current analysis provides a clinically applicable tool for guideline committees to establish evidence-based driving restrictions. PMID- 21646230 TI - Efficacy of radiofrequency catheter ablation in athletes with atrial fibrillation. AB - AIMS: Endurance sports activities have been associated with the development of atrial fibrillation (AF). Pulmonary vein isolation (PVI) by means of radiofrequency catheter ablation has been established as an effective treatment for AF. The aim of the present study was to analyse the efficacy of AF ablation in athletes. METHODS AND RESULTS: We compared procedural outcome and median term follow-up in 94 consecutive athletes (>3 h of sports/week for >= 10 years or >= 1500 h lifetime) who underwent PVI (94% men, 51 +/- 8 years, 87% paroxysmal AF, left atrial (LA) diameter 40 +/- 8 mm, mean follow-up 41 months), and 41 contemporary controls. Sixty-three per cent of athletes performed endurance sports (running, cycling, swimming, and rowing). Documented focal induction of AF and failed treatment with >= 1 anti-arrhythmic drug were pre-requisites for selection of ablation treatment. Patients with long-standing persistent or permanent AF or an LA diameter >= 55 mm were not considered for ablation. Median lifetime cumulative hours of sports was 8638 (4175-13 688) in athletes vs. 450 (280-600) in controls (P < 0.001). Other baseline characteristics except for gender (94 vs. 66% men, respectively, P < 0.001) were comparable between both groups, as was the total number of ablation procedures per patient (1.2 +/- 0.5, P = 0.62). Survival analysis showed similar AF recurrence rate after a first ablation for controls and endurance athletes, though non-endurance athletes had a significantly higher AF recurrence rate (48 vs. 46 vs. 34% freedom from AF at 3 year follow-up after a single ablation, P= 0.04). Final outcome after all ablations was similar (87 vs. 84 vs. 85% freedom from AF at 3-year follow-up, P = 0.88). No other independent predictor for AF recurrence was identified. CONCLUSION: In patients with documented focal induction of non-permanent AF and absence of structural heart disease, PVI is as effective in endurance athletes as in other patients. PMID- 21646231 TI - Intensity of continuous renal replacement therapy in acute kidney injury in the intensive care unit: a systematic review and meta-analysis. AB - BACKGROUND AND OBJECTIVE: Acute kidney injury is a common finding among patients in the intensive care unit (ICU) and is an independent predictor of mortality. The optimal intensity and timing of continuous renal replacement therapy (CRRT), in critically ill patients remain unclear. The purpose of this study was to conduct a systematic review and meta-analysis of all prospective randomized controlled trials (RCTs) to determine the effect of intensity of CRRT on the survival of patients with acute renal failure (ARF) in ICU setting. METHODS: Search strategy and data source. Electronic databases were searched on MEDLINE (through February 2010), ISIWeb of Science, and Cochrane Central Register of Controlled Trials (2010); Pub Med ''Related articles.'' Trial authors were also contacted for additional information. Study selection and data abstraction. All prospective clinical trials comparing the intensity of CRRT in adult patients with ARF and with explicit reporting of mortality were included. Three authors independently evaluated articles for eligibility and extracted data on study quality and outcomes. Meta-analysis used a random-effects model. RESULT: Of the 322 citations, 5 trials (n = 2402) were included in the meta-analysis, which met all the inclusion and exclusion criteria. Meta-analysis showed that in critically ill patients with acute kidney injury, the high-dose CRRT did not reduce mortality at 28 days. (risk ratio [RR], 0.88; 95% confidence interval [CI], 0.70 1.11; P = 0.28). CONCLUSION: In critically ill patients with acute kidney injury, the high-dose CRRT did not reduce mortality at 28 days. PMID- 21646232 TI - The use of the StarClose device for obtaining femoral artery hemostasis. AB - BACKGROUND: Following percutaneous intervention, obtaining femoral artery hemostasis quickly and reliably with few complications is of considerable importance. While the StarClose device has been studied in patients undergoing cardiac catheterization, its use in patients undergoing treatment for peripheral vascular disease has rarely been reported. It is the purpose of this report to determine whether the Starclose is able to deliver safe and effective femoral artery closure in this group of patients. METHODS: The records of all patients undergoing StarClose closure of the femoral artery by or under the supervision of a single vascular surgeon were reviewed. Indication, type of intervention, and size of the vascular sheath employed were noted. Times to mobilization and discharge were tabulated. Complications including hemorrhage, pseudoaneurysm, infection, and vessel occlusion were recorded. RESULTS: Between February 2006 and September 2009, 603 StarClose nitinol clip closure devices were deployed in 478 patients. In all, 97 procedures were diagnostic and 506 included therapeutic interventions. A total of 97 5F sheaths, 465 6F sheaths, and 41 7F sheaths were employed. Time to ambulation and discharge was 44.2 +/- 13.2 minutes and 119.3 +/ 22.6 minutes in patients in whom a 5F sheath was used, 112.5 +/- 13.5 minutes and 157.5 +/- 20.6 minutes when a 6F sheath was used, and 121.9+/-38.8 minutes and 160.2+/-43.2 minutes when a 7F sheath was employed. The clip could not be successfully deployed in 21 arteries (3.5%) and manual compression was successful in achieving hemostasis in 17 patients without complication. Three patients (0.5%) developed major hematomas requiring transfusion; 1 patient developed a pseudoaneurysm (0.17%) requiring thrombin injection. A single patient (0.17%) occluded his common femoral artery following StarClose deployment and 1 patient (0.17%) developed a femoral artery stenosis requiring balloon dilatation. No patient developed a groin infection. CONCLUSIONS: The StarClose provides a safe and reliable method of achieving femoral artery closure following percutaneous intervention for peripheral arterial disease. When successfully deployed, it allows for early ambulation and discharge. Since it remains entirely extraluminal, it offers advantages over other closure devices and can be safely used in the vast majority of patients with peripheral vascular disease. PMID- 21646233 TI - Alternative technique for endovascular treatment of an abdominal aortic aneurysm with a severely angulated neck. AB - An 86-year-old female was incidentally discovered with an asymptomatic 5.3 cm infrarenal aortic aneurysm. Preoperative imaging demonstrated the aortic neck to have severe suprarenal and infrarenal angulation. The patient was deemed unfit for open aortic repair due to multiple medical comorbidities. Endovascular treatment was performed with a modified Zenith bifurcated graft using a Zenith TX2 endograft as a proximal aortic cuff. There were no perioperative complications, and postoperative imaging demonstrated aneurysm exclusion with no migration or major endoleaks. The patient died of unrelated causes at 21 months. The use of a thoracic endograft as a proximal cuff is an alternative technique in the endovascular treatment of abdominal aortic aneurysms with a severely angulated neck. PMID- 21646234 TI - Treatment of pulmonary embolism using ultrasound-accelerated thrombolysis directly into pulmonary arteries. AB - Traditional therapy for pulmonary embolism includes systemic anticoagulation, systemic thrombolysis, catheter-directed thrombolysis / suction catheter thrombectomy, and surgical thromboembolectomy. Currently, the standard treatment for submassive and massive pulmonary embolism involves the use of systemic anticoagulation. However, unlike systemic anticoagulation there is no standard treatment algorithm for the use of thrombolytics to aggressively treat pulmonary embolism and its sequelae. This case report discusses the successful use of thrombolytics using the EKOS EkoSonic Ultrasound-Accelerated Thrombolysis System in the treatment of bilateral submassive pulmonary emboli along with a saddle pulmonary embolus. The EKOS ultrasound-accelerated thrombolysis procedure resulted in rapid substantial clinical improvement, resolution of bilateral pulmonary emboli along with resolution of the saddle pulmonary embolus, restoration of pulmonary blood flow with resolution of pulmonary hypertension, and normalization of pulmonary embolism-related cardiac dysfunction. This novel application of ultrasound-accelerated thrombolytic infusion directly into the pulmonary arteries for pulmonary embolism provides a potential new treatment option and a valuable addition to the treatment algorithm for the management of both submassive and massive pulmonary embolism. PMID- 21646235 TI - Remote ischemic preconditioning for cardiovascular surgery: an updated meta analysis of randomized trials. PMID- 21646236 TI - Carotid endarterectomy is superior to carotid angioplasty and stenting for perioperative and long-term results. AB - OBJECTIVE: Carotid angioplasty and stenting (CAS) has challenged carotid endarterectomy (CEA) as the therapy of choice for carotid disease. This meta analysis aims at summarizing the most current body of evidence. METHODS: All prospective, controlled clinical trials comparing CEA versus CAS were included. The outcome measures of interest were relative risk (RR) of 30-day stroke, 30-day stroke/death, long-term risk of stroke, and risk of restenosis. RESULTS: The RR of 30-day stroke for CAS was 1.6 times that of CEA (RR 1.6; 95%CI 1.2-2.0, P = .001). The 30-day RR of stroke/death was 1.5 times higher for CAS (RR 1.5; 95%CI 1.1-2.1, P = .008). There was a higher risk of long-term stroke (RR 1.2; 95%CI 1.0-1.5, P = .043). The risk of restenosis was twice for CAS (RR 1.8; 95%CI 1.1 3.1, P = .04). CONCLUSION: The 30-day RR of stroke, stroke/death, long-term risk of stroke, and risk of restenosis are consistently higher for carotid artery stenting (CAS). PMID- 21646237 TI - Carotid tortuosity in patients with prior cervical radiation: increased technical challenge during carotid stenting. AB - INTRODUCTION: Anatomic distortion associated with radiation-induced tissue changes may pose challenges for patients with prior cervical irradiation undergoing carotid stenting. We sought to evaluate the effect of these changes on carotid intervention. METHODS: Carotid angioplasty and stenting (CAS) for high grade stenosis was performed in 203 patients. In all, 12 consecutive patients with prior ipsilateral cervical irradiation were age-/sex-matched to 24 controls. Degree of internal carotid (IC) tortuosity was assessed by 4 methods: (a) deviation of IC from common carotid (CCA) axis, (b) number of intersections between this axis and the course of the IC, (c) total degrees of angulation along the course of the extracranial IC, and (d) the IC length to straight-line distance ratio. RESULTS: Carotid angioplasty and stenting was successful in all patients. Mean age was 72.8 +/- 10 years; 58.4% were male. Twenty-nine percent were symptomatic (14.4% transient ischemic attack [TIA], 8.5% cardiovascular accident [CVA], and 6.5% amaurosis). Comorbidities were similar between the entire cohort and the subgroups of irradiated/control patients. The IC revealed a higher degree of deviation from the axis of the CCA in the previously irradiated patients compared to those without radiation (29.2 degrees +/- 4.5 degrees vs 13.0 degrees +/- 2.0 degrees , P = .001) and was more likely to intersect this axis in those with a history of cervical irradiation (83.3% vs 14.3%, P < .05). Irradiated patients also exhibited a significantly greater degree of tortuosity versus nonirradiated patients when assessed by total angulation along the course of the carotid (171.8 degrees +/- 26.0 degrees vs 74.2 degrees +/- 20.2 degrees , P = .014) and by the IC length:distance ratio (1.14 +/- 0.05 vs 1.04 +/ 0.03, P = .020). Despite increased IC tortuosity in patients with prior irradiation, all procedures were successfully completed and there did not appear to be a predilection for a specific filter type. CONCLUSIONS: A history of cervical irradiation is associated with increased tortuosity of the IC, leading to potential challenges for filter and stent deployment. However, this increased procedural complexity did not affect technical success rate or device selection in this series. PMID- 21646238 TI - Continuous surveillance of lower limb perfusion during aortic surgery with near infrared spectroscopy: a pilot study. AB - BACKGROUND: Near-infrared spectroscopy (NIRS) is a noninvasive technique that allows continuous monitoring of the regional hemoglobin oxygen saturation (rSO(2)) index. We evaluated its application to survey perioperative lower limb perfusion. METHODS: A total of 10 patients (7 men, aged 71) were monitored during abdominal surgery for aortic aneurysms. The rSO(2) index was measured at the M gastrocnemius (optode 1) and at the dorsum of the foot (optode 2). RESULTS: Mean baseline rSO(2) values for optodes 1 and 2 were 67 and 66, respectively. After clamping the aorta or iliacofemoral arteries, rSO(2) dropped to 32 for optode 1 (P<.0001) and to 27 for optode 2 (P<.0001). After declamping, rSO(2) increased to 74 for optode 1 (P=.0012 vs baseline) and also to 74 for optode 2 (P=.0018 vs baseline). CONCLUSION: Near-infrared spectroscopy is an easily applicable, noninvasive tool for continuous surveillance of lower extremity perfusion during aortic reconstruction. PMID- 21646239 TI - Case report of a hybrid endovascular approach to an abdominal aortic dissection with retrograde thoracic extension and infrarenal aneurysm. AB - Spontaneous abdominal aortic dissection (AAD) with retrograde thoracic extension is an extremely rare occurrence with a high mortality. Abdominal aortic dissection can be associated with an abdominal aortic aneurysm (AAA) and the presence of an AAD with an AAA mandates surgical intervention because of a high rate of rupture. We present the case of a 53-year-old woman with a spontaneous AAD that extended retrograde into the thoracic aorta with a concomitant supraceliac intimal tear and an infrarenal AAA repaired electively with a hybrid approach using a supraceliac stent graft and an open infrarenal aortobiiliac graft. This hybrid approach provided an excellent outcome of this rare and complex vascular pathology. PMID- 21646240 TI - Comprehensive characterization of secreted aspartic proteases encoded by a virulence gene family in Candida albicans. AB - Candida albicans is a commensal organism, but causes life-threatening infections in immunocompromised patients. Certain factors such as yeast-hyphae transition and hydrolytic enzymes are suggested as virulence attributes of C. albicans. Among them, 10 types of secreted aspartic protease (SAP) genes have received particular attention as a major virulence gene family. However, their full functional repertoire, including its biochemical properties, remains to be elucidated. Hence, we purified all Sap isozymes using Pichia pastoris and comprehensively determined and compared their biochemical properties. While optimum pH of Sap7 was 6.5 and that of Sap8 was 2.5, presence of other Sap isozymes functioning within a broad range of optimum pH could allow C. albicans to survive and cause infections in various tissues. The substrate specificities of Sap isozymes were analysed by using FRETS-25Xaa libraries. Sap7 and Sap10 showed high substrate specificity, while other Sap isozymes had broad substrate specificities. Principal component analysis revealed that the 10 Sap isozymes were clustered into 3 distinct groups in terms of their substrate specificities. Interestingly, Sap4-6, which are coproduced in the hyphal form, were clustered as the same group, indicating that they may target similar host proteins. These results will lead to further understanding of C. albicans pathogenicity. PMID- 21646241 TI - The relationship between quilting and wellbeing. AB - BACKGROUND: Within public health, activities that promote and maintain wellbeing remain limited and relatively unexplored. In particular, little is known about the relationship between creative craft hobbies and wellbeing in the general population. METHODS: Twenty-nine semi-structured interviews were conducted with women from a local quilting group. Ideas and experiences of quilting in relation to wellbeing were explored with participants. Interviews were audio-recorded, transcribed verbatim and thematically analysed. RESULTS: Cognitive, emotional and social processes were uncovered, which participants identified as important for their wellbeing. Participants found quilting to be a productive use of time and an accessible means of engaging in free creativity. Colour was psychologically uplifting. Quilting was challenging, demanded concentration and participants maintained and learned new skills. Participants experienced 'flow' while quilting. A strong social network fostered the formation of strong friendships. Affirmation from others boosted self-esteem and increased motivation for skill development. Quilts were often given altruistically and gave quilting added purpose. CONCLUSIONS: The findings illustrate how creative craft hobbies such as quilting can be a meaningful vehicle for enhancing wellbeing. This study sets the foundation for further research into creativity, creative hobbies and hobbies in general. PMID- 21646242 TI - Is ischaemia-modified albumin a test for venous thromboembolism? AB - OBJECTIVE: Patients with symptoms of deep vein thrombosis (DVT) and pulmonary embolism (PE) commonly present to the emergency department (ED). The aim of this study was to assess the role of ischaemia-modified albumin (IMA) testing in the diagnosis of venous thromboembolism (VTE). METHODS: This was a prospective diagnostic cohort study. Inpatients and ED patients >16 years of age investigated for PE or DVT at a single hospital were eligible for study consent. Blinded IMA analysis was performed on the first blood sample taken from each patient. Patients underwent reference standard investigation for PE or DVT, including 3 month follow-up. Receiver operating characteristic (ROC) curves were constructed for IMA and the IMA:albumin ratio in the diagnosis of all VTE, PE and DVT. A sensitivity analysis was performed. RESULTS: 452 patients were consented and investigated for DVT, and 354 patients were consented and investigated for PE (806 in total). 348 patients investigated for PE had IMA testing as did 195 of the first 199 DVT patients. VTE prevalence was 19.7%. The IMA:albumin ratio performed better than IMA alone. The area under the ROC curve (AUC) for IMA:albumin in all VTE was 0.60 (95% CI 0.54 to 0.66), in DVT 0.56 (95% CI 0.46 to 0.65) and in PE 0.63 (95% CI 0.56 to 0.71). In ED patients with symptoms of PE, the AUC for IMA:albumin was 0.69 (95% CI 0.60 to 0.78). CONCLUSIONS: IMA testing cannot be used alone to diagnose DVT or PE, although there is a moderate association with PE in ED patients. PMID- 21646243 TI - Capturing paediatric injury in Ontario: differences in injury incidence using self-reported survey and health service utilisation data. AB - OBJECTIVE: Population-based health surveys are increasingly popular sources of data on injury occurrence. Self-reported surveys can yield estimates of the total incidence of non-fatal injuries while simultaneously capturing a rich repository of contextual data that may be informative for exploring determinants of injury risk. Although survey data are rarely recognised as complete, several researchers have expressed concerns about the sensitivity and validity of self-reported injury data, questioning whether captured cases are representative of the population experience of injury, particularly among children and youth. The present study sought to compare the population incidence of paediatric injury estimated from self-reported survey responses to those documented by a complete capture health service utilisation database among Ontario children. METHODS: Injury incidence rates documented from the National Longitudinal Survey of Children and Youth and the National Population Health Survey were compared with those reported in Canada's National Ambulatory Care Reporting System for Ontario youth aged 0-19 years for fiscal year 2002/3, stratified by the child's age and geographical location of residence. RESULTS: The two self-reported health surveys underestimated the population incidence of injury among Ontario children by at least 49% and 53%, respectively. Systematic errors exist in survey data capture such that injuries in infants and preschoolers (<4 years of age) and urban residents were most likely to be missed by the population health surveys. CONCLUSION: Injury incidence estimated through self-report is not representative of the population burden and experience of paediatric injury for Ontario children, and may produce biased estimates of risk when analysed as independent sources of data. PMID- 21646244 TI - Assessing the concordance of health and child protection data for 'maltreated' and 'unintentionally injured' children. AB - OBJECTIVES: To quantify the concordance of hospital child maltreatment data with child protection service (CPS) records, and identify factors associated with linkage. METHODS: Multivariable logistic regression analysis was conducted following retrospective medical record review and database linkage of 884 child records from 20 hospitals and the CPS in Queensland, Australia. RESULTS: Nearly all children with hospital assigned maltreatment codes (93.1%) had a CPS record. Of these, 85.1% had a recent notification. 29% of the linked maltreatment group (n=113) were not known to the CPS prior to the hospital presentation. Almost one third of children with unintentional injury hospital codes were known to the CPS. Just over 24% of the linked unintentional injury group (n=34) were not known to the CPS prior to the hospital presentation but became known during or after discharge from hospital. These estimates are higher than the 2006/2007 annual rate of 2.39% of children being notified to the CPS. Rural children were more likely to link to the CPS, and children were over three times more likely to link if the index injury documentation included additional diagnoses or factors affecting their health. CONCLUSIONS: The system for referring maltreatment cases to the CPS is generally efficient, although up to 1 in 15 children had codes for maltreatment but could not be linked to CPS data. The high proportion of children with unintentional injury codes who linked to CPS suggests that clinicians and hospital-based child protection staff should be supported by further education and training to ensure children at risk are being detected by the child protection system. PMID- 21646245 TI - Acute effects of the ACE inhibitor enalaprilat on the pulmonary, cerebral and systemic blood flow and resistance after the bidirectional cavopulmonary connection. AB - BACKGROUND: The bidirectional cavopulmonary connection (BCPC) is used in the staged palliation of univentricular hearts and places the cerebral and pulmonary vascular beds in series. Angiotensin-converting enzyme inhibitors (ACEI) are often used in this complex circulation, but the effects of their vasodilation are unclear. OBJECTIVE: Assessment of the acute response of perfusion pressure, flow and resistance across the systemic, cerebral and pulmonary vascular beds to ACEI in patients with a BCPC. DESIGN: Prospective interventional study. SETTING: Single tertiary care centre. PATIENTS: 12 patients with a BCPC (median age 28 months, weight 11.8 kg) undergoing a pre-Fontan catheterisation with MRI measurement of flows. INTERVENTION: Intravenous enalaprilat 0.005 or 0.01 mg/kg. RESULTS: Enalaprilat increased descending aorta flow (median 21.6%, p=0.0005), decreased total pulmonary vein flow (median 10.6%, p=0.025), and both superior caval vein flow (median 8.6%, p=0.065) and aortopulmonary collateral flow (median 15.5%, p=0.077) tended to decrease. Total cardiac output was unchanged (p=0.57). Systemic vascular resistance (median 41.9%, p=0.0005) and cerebral vascular resistance (median 23.4%, p=0.0005) decreased, but pulmonary vascular resistance (p=0.73) showed little change. There was evidence of autoregulation of cerebral blood flow. The proportion of descending aortic flow to total cardiac output increased (median 27 to 35%, p=0.001). Systemic oxygen saturation decreased from 87% to 83% (p=0.02). CONCLUSION: Enalaprilat did not increase total cardiac output but redistributed flow to the lower body, with a concomitant decrease in arterial oxygen saturation. It is difficult to increase cardiac output in patients with a BCPC and ACEI should be used with caution in those with borderline aortic saturations. PMID- 21646246 TI - Development of the first disability index for inflammatory bowel disease based on the international classification of functioning, disability and health. AB - OBJECTIVE: The impact of inflammatory bowel disease (IBD) on disability remains poorly understood. The World Health Organization's integrative model of human functioning and disability in the International Classification of Functioning, Disability and Health (ICF) makes disability assessment possible. The ICF is a hierarchical coding system with four levels of details that includes over 1400 categories. The aim of this study was to develop the first disability index for IBD by selecting most relevant ICF categories that are affected by IBD. METHODS: Relevant ICF categories were identified through four preparatory studies (systematic literature review, qualitative study, expert survey and cross sectional study), which were presented at a consensus conference. Based on the identified ICF categories, a questionnaire to be filled in by clinicians, called the 'IBD disability index', was developed. RESULTS: The four preparatory studies identified 138 second-level categories: 75 for systematic literature review (153 studies), 38 for qualitative studies (six focus groups; 27 patients), 108 for expert survey (125 experts; 37 countries; seven occupations) and 98 for cross sectional study (192 patients; three centres). The consensus conference (20 experts; 17 countries) led to the selection of 19 ICF core set categories that were used to develop the IBD disability index: seven on body functions, two on body structures, five on activities and participation and five on environmental factors. CONCLUSIONS: The IBD disability index is now available. It will be used in studies to evaluate the long-term effect of IBD on patient functional status and will serve as a new endpoint in disease-modification trials. PMID- 21646247 TI - Interleukin-1beta (IL-1beta) promotes susceptibility of Toll-like receptor 5 (TLR5) deficient mice to colitis. AB - BACKGROUND: The extent to which numerous strains of genetically engineered mice, including mice lacking Toll-like receptor 5 (T5KO), display colitis is environment dependent. Gut microbiota underlie much of the variation in phenotype. Accordingly, embryonic rederivation of T5KO mice ameliorated their spontaneous colitis despite only partially correcting elevated proinflammatory gene expression. It was postulated that endogenous anti-inflammatory pathways mediated the absence of overt inflammation in these mice when their gut microbiota were reset. Consequently, it was hypothesised that neutralisation of the anti-inflammatory cytokine interleukin 10 (IL-10) might induce uniform colitis in T5KO mice, and thus provide a practical means to study mechanisms underlying their inflammation. METHODS: Two distinct strains of non-colitic T5KO mice, as well as mice lacking MyD88, Toll-like receptor 4 (TLR4), IL-1 receptor (IL-1R) and various double knockouts (DKOs) were treated weekly for 4 weeks with 1 mg/mouse of IL-10 receptor neutralising antibody (IL-10R mAb) and colitis assayed 1 week later. The composition of the caecal microbiota was determined by 454 pyrosequencing of 16S rRNA genes. RESULTS: Anti-IL-10R mAb treatment led to severe uniform intestinal inflammation in both strains of T5KO mice. Such neutralisation of IL-10 signalling did not cause colitis in wild-type littermates nor mice lacking TLR4, MyD88 or IL-1R. The susceptibility of T5KO mice to this colitis model was not rescued by absence of TLR4 in that T4/T5 DKO mice displayed severe colitis in response to anti-IL-10R mAb treatment. IL-1beta signalling was crucial for this colitis model in that IL-1R/T5 DKOs were completely protected from colitis in response to IL-10R mAb treatment. Lastly, it was observed that blockade of IL-10R function was associated with changes in the composition of gut microbiota, which were observed in mice that were susceptible and resistant to IL 10R mAb-induced colitis. CONCLUSION: Regardless of whether they harbour a colitogenic microbiota, loss of TLR5 predisposes mice to colitis triggered by immune dysregulation via an IL-1beta-dependent pathway. PMID- 21646248 TI - Environmental transportation of tritium and estimation of site-specific model parameters for Kaiga site, India. AB - Tritium content in air moisture, soil water, rain water and plant water samples collected around the Kaiga site, India was estimated and the scavenging ratio, wet deposition velocity and ratio of specific activities of tritium between soil water and air moisture were calculated and the results are interpreted. Scavenging ratio was found to vary from 0.06 to 1.04 with a mean of 0.46. The wet deposition velocity of tritium observed in the present study was in the range of 3.3E-03 to 1.1E-02 m s(-1) with a mean of 6.6E-03 m s(-1). The ratio of specific activity of tritium in soil moisture to that in air moisture ranged from 0.17 to 0.95 with a mean of 0.49. The specific activity of tritium in plant water in this study varied from 73 to 310 Bq l(-1). The present study is very useful for understanding the process and modelling of transfer of tritium through air/soil/plant system at the Kaiga site. PMID- 21646249 TI - Adolescent pharyngitis: a review of bacterial causes. PMID- 21646250 TI - No excuse. PMID- 21646251 TI - Physician concussion knowledge and the effect of mailing the CDC's "Heads Up" toolkit. AB - BACKGROUND: The Centers for Disease Control and Prevention's (CDC) "Heads Up" toolkit was designed to educate physicians about concussion, but it has not been well studied. This study proposed to evaluate the effect of receiving the toolkit on physician concussion knowledge. METHODS: The authors obtained a sample of physicians from the American Medical Association masterfile and randomly selected half to be mailed the CDC's "Heads Up" toolkit. All physicians were then sent a survey on concussion knowledge. Data were analyzed to evaluate the effect of the toolkit on concussion knowledge. RESULTS: The survey was completed by 414 physicians (183 intervention, 231 control). There were no differences in general concussion knowledge between intervention and control groups, but physicians in the intervention group were significantly less likely to recommend next day return to play after a concussion (adjusted odds ratio = 0.31, 95% confidence interval = 0.12-0.76). CONCLUSIONS: Mailing the CDC's "Heads Up" toolkit appears to affect physicians' recommendations regarding returning to play after a concussion. PMID- 21646252 TI - Can we predict hypertension among preterm children? AB - Prematurity and low birth weight (LBW) cause hypertension (HTN), ischemic heart disease, and obesity in young adults. The objective of the study was to identify risk factors for the development of HTN in children born preterm or at a LBW and to assess pediatricians' awareness of the problem. A retrospective review of 160 cases was conducted. In total, 22% of babies born preterm/LBW developed HTN by age 15 years. The odds of developing HTN were 1.6 times greater for every one standard deviation increase in body mass index. Higher risk posses for those born small for gestational age and under 1000 g. Of the 35 cases of HTN identified, only 31% were recognized as abnormal by the primary care providers. The development of obesity and HTN appear related in preterm/LBW children. Awareness of prematurity or LBW as a risk factor for HTN should be raised among pediatric primary care providers. PMID- 21646253 TI - Treating childhood obesity in primary care. AB - The objective of this prospective, cohort study was to assess an intervention for obese children (9-12 years of age) and their families delivered in primary care. A family-based, behavioral weight management program consisted of 11 sessions. The treatment consisted of a calorie goal, self-monitoring of daily food intake, physical activity and sedentary behavior, and other behavior change skills. A total of 78 children and families entered treatment; 23 children served as quasi controls. The mean weight loss at 15 weeks among 55 children (71%) who completed the program was 2.4 lbs (SD = 5.24, range of -16.7 to +8.4 lbs) compared with a mean weight gain of 3.45 lbs (SD = 4.31, range of -5.0 to +12.0 lbs) among 23 control children. The mean change in body mass index z score from baseline to month 24 was -0.17 +/- 0.32 (P < .001). Primary care is an appropriate place to identify and treat children with obesity. PMID- 21646254 TI - Percutaneous thoracic duct embolization as a treatment for intrathoracic chyle leaks in infants. AB - Chylothorax is an uncommon complication of cardiothoracic surgery in children that is traditionally treated with either conservative (diet modification, octreotide administration, and percutaneous drainage) or surgical (thoracic duct ligation, pleurodesis, and pleuroperitoneal shunt) approaches. We report here the cases of 2 children (a 6-month-old and a 1-month-old) with postoperative chylous leaks who were treated successfully by percutaneous thoracic duct embolization. PMID- 21646255 TI - Child abuse pediatrics: new specialty, renewed mission. PMID- 21646256 TI - New insights about infant and toddler skin: implications for sun protection. AB - The skin is increasingly recognized as a component of the innate immune response, in addition to its role as a physical barrier. Although the deleterious effects of solar ultraviolet radiation (UVR), including immunosuppression and cutaneous tumorigenesis, are widely acknowledged, most studies to date have concentrated on adult skin. Despite the more sensitive nature of infant and toddler skin, little is known about its responses to UVR exposure, whether acute or long-term. Accumulating evidence suggests not only that the skin's barrier protection remains immature throughout at least the first 2 years of life but also that accumulation of UVR-induced changes in the skin may begin as early as the first summer of life. Such evidence not only affirms the importance of sun protection during the infant and toddler years but underscores the need for more research to establish evidence-based standards of care in this area. In this article we review recent studies in which differences between the skin properties of infants and young children and those of adults were compared, and we discuss the implications of these differences for sun-protection practices. PMID- 21646257 TI - Children with asthma hospitalized with seasonal or pandemic influenza, 2003-2009. AB - OBJECTIVE: To describe the characteristics and clinical courses of asthmatic children hospitalized with seasonal or 2009 pandemic H1N1 influenza and compare complications by influenza type. METHODS: During the 2003-2009 influenza seasons and the 2009 pandemic, we conducted surveillance of 5.3 million children aged 17 years or younger for hospitalization with laboratory-confirmed influenza and identified those with asthma (defined as those aged 2-17 years with a history of asthma in their medical record or a discharge code for acute asthma exacerbation or status asthmaticus). We collected data from medical records on medical history and clinical course; data on asthma severity and control were not routinely collected. RESULTS: During the 2003-2009 influenza seasons, 701 (32%) of 2165 children hospitalized with influenza had asthma; during the 2009 pandemic, 733 (44%) of 1660 children had asthma. The median age of the asthmatic children was 7 years, and 73% had no additional medical conditions. Compared with asthmatic children with seasonal influenza, a higher proportion with 2009 pandemic H1N1 influenza required intensive care (16% vs 22%; P=.01) and were diagnosed with pneumonia (40% vs 46%; P=.04), whereas equal proportions had respiratory failure (5% vs 5%; P=.8) and died (1% vs 1%; P=.4). More asthmatic children with influenza A (seasonal or pandemic) had diagnoses of asthma exacerbations compared with those with influenza B (51% vs 29%; P<.01). CONCLUSIONS: The majority of asthmatic children hospitalized with influenza have no additional medical conditions. Complications such as pneumonia and need for intensive care occur in a substantial proportion, highlighting the importance of influenza prevention through vaccination among asthmatic children. PMID- 21646258 TI - Hemophagocytic lymphohistiocytosis with Munc13-4 mutation: a cause of recurrent fatal hydrops fetalis. AB - Familial hemophagocytic lymphohistiocytosis (FHL) is an autosomal recessive disorder of immune regulation that leads to a hyperinflammatory syndrome responsible for fever, hepatosplenomegaly, cytopenia, and coagulopathy. Although presentation usually occurs in early infancy, antenatal presentation is extremely rare. To our knowledge, we are first to report genetically confirmed FHL in 2 consecutive siblings who presented with hydrops fetalis that led to spontaneous intrauterine death at 38 and 30 weeks of gestation. Because the diagnosis of FHL has important implications for genetic counseling, we suggest that FHL be considered in the differential diagnosis of nonimmune hydrops fetalis. PMID- 21646259 TI - Instruments to detect alcohol and other drug misuse in the emergency department: a systematic review. AB - CONTEXT: Alcohol and other drug (AOD) misuse by youth is a significant public health concern. Unanticipated treatment for AOD-related morbidities is often sought in hospital emergency departments (EDs). Screening instruments that rapidly identify patients who require further diagnostic evaluation and/or brief intervention are critically important. OBJECTIVE: To summarize evidence on screening instruments that can assist emergency care clinicians in identifying AOD misuse in pediatric patients. METHODS: Fourteen electronic databases (including Medline, Embase, and PsycINFO) and reference screening were used. Psychometric and prospective diagnostic studies were selected if the instrument focused on detecting AOD misuse in patients aged 21 years or younger in the ED. Two reviewers independently assessed quality and extracted data. Validity and reliability data were collected for psychometric studies. Instrument performance was assessed by using sensitivity, specificity, and positive (LR(+)) and negative (LR(-)) likelihood ratios. Meta-analysis was not possible because of clinical and measurement heterogeneity. RESULTS: Of the 1545 references initially identified, 6 studies met inclusion criteria; these studies evaluated 11 instruments for universal or targeted screening of alcohol misuse. Instruments based on diagnostic criteria for AOD disorders were effective in detecting alcohol abuse and dependence (sensitivity: 0.88; specificity: 0.90; LR(+): 8.80) and cannabis use disorder (sensitivity: 0.96; specificity: 0.86; LR(+): 6.83). CONCLUSIONS: On the basis of the current evidence, we recommend that emergency care clinicians use a 2-question instrument for detecting youth alcohol misuse and a 1-question instrument for detecting cannabis misuse. Additional research is required to definitively answer whether these tools should be used as targeted or universal screening approaches in the ED. PMID- 21646260 TI - Sexual history documentation in adolescent emergency department patients. AB - OBJECTIVES: To determine the frequency of physician-documented sexual histories in female adolescents who presented to an emergency department (ED) with symptoms suggestive of a sexually transmitted infection (STI). Our secondary objectives were to determine if physician-documented sexual history is associated with increased STI testing and to compare the concordance of physician-elicited sexual histories with patient-documented sexual histories by using confidential questionnaires. METHODS: We conducted a secondary analysis of a prospective study of female adolescents who presented to a pediatric ED with chief complaints of lower abdominal pain and/or genitourinary complaints. Patient charts were abstracted for the presence or absence of documented sexual histories and demographics. A subset of patients completed a questionnaire pertaining to sexual health. RESULTS: The study population comprised 327 patients; 269 (82%) patients had a physician-documented sexual history, of which 204 (76%) reported being sexually active to the physician. Patient age (odds ratio [OR]: 2.6 [95% confidence interval (CI): 1.3-5.3]) and black race (OR: 2.0 [95% CI: 1.1-3.7]) were associated with physician-documented sexual history. The documentation of a sexual history was associated with increased STI testing (OR: 3.9 [95% CI: 2.0 7.6]). In the patients (n=109) who completed the questionnaire, physician elicited sexual histories were highly concordant with patient-documented sexual histories on questionnaire (Spearman r=0.90; P<.001). CONCLUSION: These results indicate that ED physicians should obtain sexual histories from symptomatic patients, because it may increase STI testing and subsequent detection. In future studies factors should be evaluated that affect physicians' willingness to assess sexual history in the ED patient. PMID- 21646261 TI - Behavior outbursts, orofacial dyskinesias, and CSF pleocytosis in a healthy child. AB - Anti-N-methyl-D-aspartate (NMDA) receptor encephalitis represents a new category of immune-mediated neurologic disorders. Viral encephalitis is often the presumptive diagnosis because of the acute neurologic changes, cerebrospinal fluid lymphocytic pleocytosis, and occasional hyperthermia. We report here the case of a previously healthy 7-year-old boy with new-onset aggressive behavior, seizure activity, and orofacial dyskinesias with cerebrospinal fluid and serum that tested positive for anti-NMDA receptors. PMID- 21646262 TI - Low-dose, high-frequency CPR training improves skill retention of in-hospital pediatric providers. AB - OBJECTIVE: To investigate the effectiveness of brief bedside cardiopulmonary resuscitation (CPR) training to improve the skill retention of hospital-based pediatric providers. We hypothesized that a low-dose, high-frequency training program (booster training) would improve CPR skill retention. PATIENTS AND METHODS: CPR recording/feedback defibrillators were used to evaluate CPR quality during simulated arrest. Basic life support-certified, hospital-based providers were randomly assigned to 1 of 4 study arms: (1) instructor-only training; (2) automated defibrillator feedback only; (3) instructor training combined with automated feedback; and (4) control (no structured training). Each session (time: 0, 1, 3, and 6 months after training) consisted of a pretraining evaluation (60 seconds), booster training (120 seconds), and a posttraining evaluation (60 seconds). Excellent CPR was defined as chest compression (CC) depth >= one-third anterior-posterior chest depth, rate >=90 and <=120 CC per minute, <=20% of CCs with incomplete release (>2500 g), and no flow fraction <= 0.30. MEASUREMENTS AND MAIN RESULTS: Eighty-nine providers were randomly assigned; 74 (83%) completed all sessions. Retention of CPR skills was 2.3 times (95% confidence interval [CI]: 1.1-4.5; P=.02) more likely after 2 trainings and 2.9 times (95% CI: 1.4 6.2; P=.005) more likely after 3 trainings. The automated defibrillator feedback only group had lower retention rates compared with the instructor-only training group (odds ratio: 0.41 [95% CI: 0.17-0.97]; P = .043). CONCLUSIONS: Brief bedside booster CPR training improves CPR skill retention. Our data reveal that instructor-led training improves retention compared with automated feedback training alone. Future studies should investigate whether bedside training improves CPR quality during actual pediatric arrests. PMID- 21646263 TI - Neonatal Graves' disease associated with severe metabolic abnormalities. AB - Neonatal Graves' disease is a rare condition that is sometimes associated with multisystem abnormalities that can mimic infection or inborn errors of metabolism. Here we describe the cases of 2 infants who had serious metabolic derangements including conjugated hyperbilirubinemia and hyperammonemia. PMID- 21646265 TI - Academy of pediatric education and leadership: preparing leaders for educational innovation. AB - Faculty development has been identified as a critical need if we hope to advance competency-based education. Despite the small number of participants at a single institution, the model presented here has the possibility for making a significant contribution to future faculty development initiatives for two reasons. First, by supporting the time of the scholars, the program makes a statement about the value of medical education and the recognition that it requires a skill set to become an educator. While the need for requisite skill sets has long been recognized for training sub-specialists, this has not been the case for those taking on major roles in education and training. Second, despite a rich literature, little has been published about the effects on the learners of those who participated in the faculty development programs. By supporting a cadre of individuals to acquire the skills needed to be an educator as well as the skills needed to perform educational research to study the impact of applying those skills, this project serves as a model for developing a much needed community of medical education leaders. PMID- 21646266 TI - Rates of parent-centered developmental screening: disparities and links to services access. AB - BACKGROUND: In 2006, the American Academy of Pediatrics recommended developmental screening of young children with a standardized screening tool as a routine component of well-child care. OBJECTIVES: To assess the national and state prevalence of standardized, parent-completed developmental screening (DS-PC) in the previous 12 months and evaluate associations between screening and receipt of an early-intervention plan or mental health services for children at higher risk. METHODS: Data from the 2007 National Survey of Children's Health were used. Nested t tests were used to compare each state to national prevalence. Logistic and multilevel regression models evaluated variations and associations with DS PC. RESULTS: Nationally, 19.5% of children received a DS-PC in the previous 12 months, although the figure varied from 10.7% to 47% across the United States. Prevalence did not rise above 26.7% for any socioeconomic subgroup of children and was highest for younger, black, and publicly insured children and lowest for uninsured children and children with gaps in insurance coverage. Equally high risk children varied twofold in their probability of receiving early intervention or needed mental health services according to whether they had received a DS-PC. CONCLUSIONS: There is a significant gap between the developmental screening that is recommended and what is reported nationally. When children are not screened consistently, opportunities for early identification, intervention, and treatment may be delayed. Gaps in screening and wide variations across states present considerable opportunities for cross-state learning to improve quality on this critical component of preventive pediatric care. Measurement systems for assessing prevalence and impact of screening require continued evaluation and development. PMID- 21646264 TI - Targeting inflammation to prevent bronchopulmonary dysplasia: can new insights be translated into therapies? AB - Bronchopulmonary dysplasia (BPD) frequently complicates preterm birth and leads to significant long-term morbidity. Unfortunately, few therapies are known to effectively prevent or treat BPD. Ongoing research has been focusing on potential therapies to limit inflammation in the preterm lung. In this review we highlight recent bench and clinical research aimed at understanding the role of inflammation in the pathogenesis of BPD. We also critically assess currently used therapies and promising developments in the field. PMID- 21646268 TI - New applications of the renormalization group method in physics: a brief introduction. AB - The renormalization group (RG) method developed by Ken Wilson more than four decades ago has revolutionized the way we think about problems involving a broad range of energy scales such as phase transitions, turbulence, continuum limits and bifurcations in dynamical systems. The Theme Issue provides articles reviewing recent progress made using the RG method in atomic, condensed matter, nuclear and particle physics. In the following, we introduce these articles in a way that emphasizes common themes and the universal aspects of the method. PMID- 21646269 TI - Renormalization group for non-relativistic fermions. AB - A brief introduction is given to the renormalization group for non-relativistic fermions at finite density. It is shown that Landau's theory of the Fermi liquid arises as a fixed point (with the Landau parameters as marginal couplings) and its instabilities as relevant perturbations. Applications to related areas, nuclear matter, quark matter and quantum dots, are briefly discussed. The focus will be on explaining the main ideas to people in related fields, rather than addressing the experts. PMID- 21646270 TI - Renormalization group aspects of graphene. AB - Graphene is a two-dimensional crystal of carbon atoms with fascinating electronic and morphological properties. The low-energy excitations of the neutral, clean system are described by a massless Dirac Hamiltonian in (2+1) dimensions, which also captures the main electronic and transport properties. A renormalization group analysis sheds light on the success of the free model: owing to the special form of the Fermi surface that reduces to two single points in momentum space, short-range interactions are irrelevant and only gauge interactions such as long range Coulomb or effective disorder can play a role in the low-energy physics. We review these features and briefly discuss other aspects related to disorder and to the bilayer material along the same lines. PMID- 21646271 TI - The density-matrix renormalization group: a short introduction. AB - The density-matrix renormalization group (DMRG) method has established itself over the last decade as the leading method for the simulation of the statics and dynamics of one-dimensional strongly correlated quantum lattice systems. The DMRG is a method that shares features of a renormalization group procedure (which here generates a flow in the space of reduced density operators) and of a variational method that operates on a highly interesting class of quantum states, so-called matrix product states (MPSs). The DMRG method is presented here entirely in the MPS language. While the DMRG generally fails in larger two-dimensional systems, the MPS picture suggests a straightforward generalization to higher dimensions in the framework of tensor network states. The resulting algorithms, however, suffer from difficulties absent in one dimension, apart from a much more unfavourable efficiency, such that their ultimate success remains far from clear at the moment. PMID- 21646272 TI - The renormalization group and nuclear forces. AB - I give an outline of recent applications of the renormalization group to effective theories of nuclear forces, focusing on the use of a Wilsonian approach to analyse systems of two or three non-relativistic particles. PMID- 21646273 TI - Efimov physics from a renormalization group perspective. AB - We discuss the physics of the Efimov effect from a renormalization group viewpoint using the concept of limit cycles. Furthermore, we discuss recent experiments providing evidence for the Efimov effect in ultracold gases and its relevance for nuclear systems. PMID- 21646274 TI - Lattice studies of quantum chromodynamics-like theories with many fermionic degrees of freedom. AB - I give an elementary introduction to the study of gauge theories coupled to fermions with many degrees of freedom. Besides their intrinsic interest, these theories are candidates for non-perturbative extensions of the Higgs sector of the standard model. While related to quantum chromodynamics, these systems can exhibit very different behaviour from it: they can possess a running gauge coupling with an infrared-attractive fixed point. I briefly survey recent lattice work in this area. PMID- 21646275 TI - Quark confinement and the renormalization group. AB - Recent approaches to quark confinement are reviewed, with an emphasis on their connection to renormalization group (RG) methods. Basic concepts related to confinement are introduced: the string tension, Wilson loops and Polyakov lines, string breaking, string tension scaling laws, centre symmetry breaking and the deconfinement transition at non-zero temperature. Current topics discussed include confinement on R(3)*S(1), the real-space RG, the functional RG and the Schwinger-Dyson equation approach to confinement. PMID- 21646276 TI - Renormalization group flow equations with full momentum dependence. AB - After a short elementary introduction to the exact renormalization group for the effective action, I discuss a particular truncation of the hierarchy of flow equations that allows for the determination of the full momentum of the n-point functions. Applications are then briefly presented, to critical O(N) models, to Bose-Einstein condensation and to finite-temperature field theory. PMID- 21646277 TI - Renormalization group and the Planck scale. AB - I discuss the renormalization group approach to gravity, and its link to Weinberg's asymptotic safety scenario, and give an overview of results with applications to particle physics and cosmology. PMID- 21646278 TI - Functional renormalization for the Bardeen-Cooper-Schrieffer to Bose-Einstein condensation crossover. AB - We review the functional renormalization group (RG) approach to the Bardeen Cooper-Schrieffer to Bose-Einstein condensation (BCS-BEC) crossover for an ultracold gas of fermionic atoms. Formulated in terms of a scale-dependent effective action, the functional RG interpolates continuously between the atomic or molecular microphysics and the macroscopic physics on large length scales. We concentrate on the discussion of the phase diagram as a function of the scattering length and the temperature, which is a paradigm example for the non perturbative power of the functional RG. A systematic derivative expansion provides for both a description of the many-body physics and its expected universal features as well as an accurate account of the few-body physics and the associated BEC and BCS limits. PMID- 21646279 TI - Pregnancy outcome after oocyte donation in patients with Turner's syndrome and partial X monosomy. AB - BACKGROUND: Fertility expectations for patients with Turner's syndrome (TS) have clearly changed in the last three decades. However, medical risks during pregnancy are supposed to be highly increased. The aim of the study was to assess clinical outcome and obstetrical complications in a series of patients with TS in an oocyte donor programme. METHODS: A retrospective study was carried out on 24 women with TS seeking a pregnancy in the Fertility Clinic of the Erasme Hospital from 1992 up until March 2011. RESULTS: Twenty-three patients with TS were included in an oocyte donation cycle. Forty-nine oocyte donation cycles were performed, which led to 45 fresh and 10 frozen-thawed embryo transfers. Altogether, 18 pregnancies were obtained, 10 deliveries (9 singletons and 1 pair of twins), 3 miscarriages and 5 biochemical pregnancies. The clinical pregnancy rate per transfer was 24.4% in fresh cycles and 20% in frozen replacement cycles. Complications of pregnancy occurred in 5 of 10 pregnancies (50%), which led to three premature deliveries because of pregnancy-induced hypertensive disorders. The mean birthweight (g) (+/-SD) for singletons and twins was 2728 +/- 577 and 2335 +/- 318, respectively. Four babies were below the 10th percentile. No cardiac complications were observed in any of the pregnant women. CONCLUSIONS: Pregnancy rates after oocyte donation in patients with TS are comparable with those previously published but a high risk of pregnancy hypertensive disorders and a high risk of low birthweight can be highlighted from our study. Strict inclusion criteria and single embryo transfer are necessary to minimize complications during pregnancy in this high-risk group. PMID- 21646280 TI - Increased frequency of occult fragile X-associated primary ovarian insufficiency in infertile women with evidence of impaired ovarian function. AB - BACKGROUND: The FMR1 premutation is associated with overt primary ovarian insufficiency (POI). However, its prevalence in women with occult POI (i.e. menstrual cycles, but impaired ovarian response) has not been examined. We hypothesized that both the FMR1 premutation and intermediate allele is more frequent in infertile women with occult POI than in controls, and that a repeat length cutoff might predict occult POI. METHODS: All subjects were menstruating women <42 years old and with no family history of unexplained mental retardation, autism or fragile X syndrome. Cases had occult POI defined by elevated FSH or poor response to gonadotrophin therapy (n = 535). Control subjects (n = 521) had infertility from other causes or were oocyte donors. Prevalence of the FMR1 premutation and intermediate alleles was examined and allele length was compared between controls and women with occult POI. RESULTS: The frequency of the premutation (7/535 versus 1/521; P< 0.05) and intermediate alleles (17/535 versus 7/521; P< 0.05) was higher in women with occult POI than in controls. The allele with the greatest number of CGG repeats was longer in women with occult POI compared with controls (32.7 +/- 7.1 versus 31.6 +/- 4.3; P < 0.01). A receiver operating characteristic curve examining repeat length as a test for occult POI had an area of 0.56 +/- 0.02 (P < 0.01). A repeat cutoff of 45 had a specificity of 98%, but a sensitivity of only 5% to identify occult POI. The positive predictive value was only 21% for a fertility population that has ~ 22% of its patients with occult POI. CONCLUSIONS: The data suggest that FMR1 premutations and intermediate alleles are increased in women with occult POI. Thus, FMR1 testing should be performed in these women as some will have fragileX-associated POI. Although the FMR1 repeat lengths were longer in women with occult POI, the data do not support the use of a repeat length cutoff to predict occult POI. PMID- 21646281 TI - Leaf-trait variation explained by the hypothesis that plants maximize their canopy carbon export over the lifespan of leaves. AB - Measured values of four key leaf traits (leaf area per unit mass, nitrogen concentration, photosynthetic capacity, leaf lifespan) co-vary consistently within and among diverse biomes, suggesting convergent evolution across species. The same leaf traits co-vary consistently with the environmental conditions (light intensity, carbon-dioxide concentration, nitrogen supply) prevailing during leaf development. No existing theory satisfactorily explains all of these trends. Here, using a simple model of the carbon-nitrogen economy of trees, we show that global leaf-trait relationships and leaf responses to environmental conditions can be explained by the optimization hypothesis (MAXX) that plants maximize the total amount of carbon exported from their canopies over the lifespan of leaves. Incorporating MAXX into larger-scale vegetation models may improve their consistency with global leaf-trait relationships, and enhance their ability to predict how global terrestrial productivity and carbon sequestration respond to environmental change. PMID- 21646282 TI - Free radical scavenging (DPPH) potential in nine Mentha species. AB - Mentha species are used in every day life in various food items. These species produce valuable secondary metabolites that scavenge toxic free radicals. Toxic free radicals can cause different diseases in the human body. In the present study free radical scavenging potential (1,1-diphenyl-2-picrylhydrazyl scavenging activity) in nine Mentha species were investigated to evaluate and explore new potential sources for natural antioxidants. The activity was performed after different time intervals with incubation period of 30 minutes. The methanolic extracts revealed that significantly higher activity (82%) was observed in Mentha suaveolens, followed by Mentha longifolia (79%), Mentha officinalis (76%) and Mentha piperita, Mentha pulegium, Mentha royleana (75%), respectively. Significantly same activity was observed in Mentha arvensis and Mentha spicata. Lower activity was observed in Mentha citrata (64%). The present study revealed that these species can be used as natural antioxidants. PMID- 21646283 TI - Liraglutide as additional treatment for type 1 diabetes. AB - OBJECTIVE: To determine whether the addition of liraglutide to insulin to treat patients with type 1 diabetes leads to an improvement in glycemic control and diminish glycemic variability. SUBJECTS AND METHODS: In this study, 14 patients with well-controlled type 1 diabetes on continuous glucose monitoring and intensive insulin therapy were treated with liraglutide for 1 week. Of the 14 patients, eight continued therapy for 24 weeks. RESULTS: In all the 14 patients, mean fasting and mean weekly glucose concentrations significantly decreased after 1 week from 130+/-10 to 110+/-8 mg/dl (P<0.01) and from 137.5+/-20 to 115+/-12 mg/dl (P<0.01) respectively. Glycemic excursions significantly improved at 1 week. The mean s.d. of glucose concentrations decreased from 56+/-10 to 26+/-6 mg/dl (P<0.01) and the coefficient of variation decreased from 39.6+/-10 to 22.6+/-7 (P<0.01). There was a concomitant fall in the basal insulin from 24.5+/ 6 to 16.5+/-6 units (P<0.01) and bolus insulin from 22.5+/-4 to 15.5+/-4 units (P<0.01). In patients who continued therapy with liraglutide for 24 weeks, mean fasting, mean weekly glucose concentrations, glycemic excursions, and basal and bolus insulin dose also significantly decreased (P<0.01). HbA1c decreased significantly at 24 weeks from 6.5 to 6.1% (P=0.02), as did the body weight by 4.5+/-1.5 kg (P=0.02). CONCLUSION: Liraglutide treatment provides an additional strategy for improving glycemic control in type 1 diabetes. It also leads to weight loss. PMID- 21646284 TI - Relationship of CYP21A2 genotype and serum 17-hydroxyprogesterone and cortisol levels in a large cohort of Italian children with premature pubarche. AB - OBJECTIVE: Premature pubarche (PP) is the most frequent sign of nonclassic congenital adrenal hyperplasia (NCCAH) due to 21-hydroxylase deficiency in childhood. The aim of this study was to assess the relationship between the CYP21A2 genotype and baseline and ACTH-stimulated 17-hydroxyprogesterone (17-OHP) and cortisol serum levels in patients presenting with PP. PATIENTS AND METHODS: A total of 152 Italian children with PP were studied. Baseline and ACTH-stimulated 17-OHP and cortisol serum levels were measured and CYP21A2 gene was genotyped in all subjects. RESULTS: Baseline and ACTH-stimulated serum 17-OHP levels were significantly higher in NCCAH patients than in both heterozygotes and children with idiopathic PP (IPP). Of the patient population, four NCCAH patients (7.3%) exhibited baseline 17-OHP values <2 ng/ml (6 nmol/l). An ACTH-stimulated 17-OHP cutoff level of 14 ng/ml (42 nmol/l) identified by the receiver-operating characteristics curves showed the best sensitivity (90.9%) and specificity (100%) in distinguishing NCCAH patients. This value, while correctly identifying all unaffected children, missed 9% of affected individuals. Cortisol response to ACTH stimulation was <18.2 MUg/dl (500 nmol/l) in 14 NCCAH patients (28%) and none of the heterozygotes or IPP children. Among the 55 NCCAH patients, 54.5% were homozygous for mild CYP21A2 mutations, 41.8% were compound heterozygotes for one mild and one severe CYP21A2 gene mutations, and 3.6% had two severe CYP21A2 gene mutations. CONCLUSION: In children with PP, baseline 17-OHP levels are not useful to rule out the diagnosis of NCCAH, which is accomplished by means of ACTH testing only. The different percentages of severe and mild CYP21A2 gene mutations found in PP children compared with adult NCCAH patients is an indirect evidence that the enzyme defect is under-diagnosed in childhood, and it might not lead to the development of hyperandrogenic symptoms in adulthood. Stress-dose glucocorticoids should be considered in patients with suboptimal cortisol response to ACTH stimulation. PMID- 21646285 TI - Assessment of primary cancers in GH-treated adult hypopituitary patients: an analysis from the Hypopituitary Control and Complications Study. AB - OBJECTIVE: GH and IGFs have mitogenic properties, causing speculation that GH treatment could increase risk of malignancy. While studies in GH-treated childhood cancer survivors have suggested a slight increase in second neoplasms, studies in GH-treated adults have been equivocal. Design Incidence of de novo and second cancers was evaluated in 6840 GH-treated and 940 non GH-treated adult patients in the Hypopituitary Control and Complications Study pharmacoepidemiological database. METHODS: Evident cancer cases were evaluated in the main analysis, with sensitivity analyses including probable and possible cancers. Standardized incidence ratios (SIRs) for cancers were calculated using Surveillance, Epidemiology and End Results for the USA and GLOBOCAN for all other countries. RESULTS: During the mean follow-up of 3.7 years/GH-treated patient, 142 evident cancer cases were identified, giving an overall SIR of 0.88 (95% confidence interval (CI) 0.74-1.04); 95% CIs included the value of 1.0 for each country examined. The SIR for GH-treated patients from the USA (71 cases) was 0.94 (95% CI 0.73-1.18), and for non GH-treated patients from the USA (27 cases) was 1.16 (95% CI 0.76-1.69). For GH-treated patients from the USA aged <35 years, the SIR (six cases) was 3.79 (1.39-8.26), with SIR not elevated for all other age categories; SIR for patients from the USA with childhood onset (CO) GH deficiency (GHD) was 2.74 (95% CI 1.18-5.41). The SIR for colorectal cancer in GH-treated patients (11 cases) was 0.60 (95% CI 0.30-1.08). CONCLUSIONS: With relatively short follow-up, the overall primary cancer risk in 6840 patients receiving GH as adults was not increased. Elevated SIRs were found for subgroups in the USA cohort defined by age <35 years or CO GHD. PMID- 21646286 TI - Low prevalence of hypopituitarism after traumatic brain injury: a multicenter study. AB - OBJECTIVE: Hypopituitarism after traumatic brain injury (TBI) is considered to be a prevalent condition. However, prevalence rates differ considerably among reported studies, due to differences in definitions, endocrine assessments of hypopituitarism, and confounding factors, such as timing of evaluation and the severity of the trauma. Aim To evaluate the prevalence of hypopituitarism in a large cohort of TBI patients after long-term follow-up using a standardized endocrine evaluation. Study design Cross-sectional study. PATIENTS AND METHODS: We included 112 patients with TBI, hospitalized for at least 3 days and duration of follow-up >1 year after TBI from five (neurosurgical) referral centers. Evaluation of pituitary function included fasting morning hormone measurements and insulin tolerance test (n=90) or, when contraindicated, ACTH stimulation and/or CRH stimulation tests and a GH releasing hormone-arginine test (n=22). Clinical evaluation included quality of life questionnaires. RESULTS: We studied 112 patients (75 males), with median age 48 years and mean body mass index (BMI) 26.7+/-4.8 kg/m(2). Mean duration of hospitalization was 11 (3-105), and 33% of the patients had a severe trauma (Glasgow Coma Scale <9) after TBI. The mean duration of follow-up was 4 (1-12) years. Hypopituitarism was diagnosed in 5.4% (6/112) of patients: severe GH deficiency (n=3), hypogonadism (n=1), adrenal insufficiency (n=2). Patients diagnosed with pituitary insufficiency had significantly higher BMI (P=0.002). CONCLUSION: In this study, the prevalence of hypopituitarism during long-term follow-up after TBI was low. Prospective studies are urgently needed to find reliable predictive tools for the identification of patients with a significant pre-test likelihood for hypopituitarism after TBI. PMID- 21646287 TI - Liver iron overload is associated with elevated SHBG concentration and moderate hypogonadotrophic hypogonadism in dysmetabolic men without genetic haemochromatosis. AB - AIMS: To assess the relation between moderate iron overload on sex hormone binding globulin (SHBG) levels and gonadotroph function in men with dysmetabolic iron overload syndrome and the effects of phlebotomy. METHODS: The relationship between magnetic resonance imaging assessed liver iron concentration (LIC) and plasma ferritin levels with total testosterone, bioavailable testosterone (BT), SHBG and LH levels, were studied in 50 men with moderate dysmetabolic iron excess, in the absence of genetic haemochromatosis, who were randomised to phlebotomy therapy or to normal care. RESULTS: Four patients (8%) had low total testosterone (<10.4 nmol/l) and 13 patients (26%) had low BT (<2.5 nmol/l). In the entire population, those with LIC above the median (90 MUmol/l) had a higher mean SHBG (P=0.028), lower LH (P=0.039) than those with LIC below the median. In multivariable analysis (adjusted for age, and fasting insulin) LIC was significantly associated with SHBG (positively) and LH (negatively). Patients in the highest quartile of SHBG had higher LIC (P=0.010) and higher ferritinaemia (P=0.012) than those in the three other quartiles. Iron depletion by venesection did not significantly improve any hormonal levels. CONCLUSIONS: Hypogonadism is not infrequent in men with dysmetabolic iron overload syndrome. Liver iron excess is associated with increased plasma SHBG and moderate hypogonadotrophic hypogonadism. Phlebotomy therapy needs further investigation in symptomatic hypogonadal men with dysmetabolic iron excess. PMID- 21646288 TI - Unacylated ghrelin is associated with changes in body composition and body fat distribution during long-term exercise intervention. AB - OBJECTIVE: Ghrelin, a gut-brain peptide involved in energy homeostasis, circulates predominantly (>90%) in unacylated form. Previous studies, however, have focused on total and acylated ghrelin, and the role of unacylated ghrelin (UAG) is not well understood. Particularly, the association of UAG with weight loss and changes in body composition in adults remains unclear. We hypothesized that exercise-associated increase in UAG level is associated with weight loss, favorable changes in body composition, and body fat distribution. DESIGN AND METHODS: A prospective study of 552 young men (mean age 19.3 and range 19-28 years) undergoing military service with structured 6-month exercise training program. Exercise performance, body composition, and biochemical measurements were obtained at baseline and follow-up. Association between changes in UAG levels and body composition and body fat distribution were evaluated. RESULTS: An increase in UAG level during the exercise intervention was associated with reduced weight, fat mass (FM), fat percentage (fat %), and waist circumference, but not with fat-free mass. Inverse associations of changes in UAG level with changes in waist circumference and fat % were independent of weight at baseline, and changes in weight and exercise performance. Associations of changes in UAG level with waist circumference were significantly stronger than with fat % after the adjustment for confounding variables. CONCLUSION: UAG is associated with changes in body weight and body composition during an intensive long-term exercise intervention in young men. The association of UAG levels with changes in central obesity was stronger than with total FM. PMID- 21646289 TI - Point of controversy: perioperative care of patients undergoing pheochromocytoma removal-time for a reappraisal? AB - Adrenalectomy for pheochromocytoma is reported with a mortality close to zero in recent studies. The dogma of preoperative fluid and hypotensive drug administrations is widely applied in patients scheduled for pheochromocytoma removal and is assumed to have a beneficial effect on operative outcomes. This paradigm is only based on historical studies of non-standardized practices and criteria for efficacy, with no control group. Pre- and intraoperative hypovolemia have never been demonstrated in patients scheduled for pheochromocytoma removal. Recent improvements in outcome of patients undergoing adrenalectomy for pheochromocytoma could also be the result of improvement in surgical techniques and refinement in anesthetic practices. Whether better knowledge of the disease, efficiency of available intravenous short-acting vasoactive drugs, and careful intraoperative handling of the tumor make it possible to omit preoperative preparation in most patients scheduled for pheochromocytoma removal is presently questionable. We reviewed available literature in this respect. PMID- 21646290 TI - Novel inactivating mutations in the GH secretagogue receptor gene in patients with constitutional delay of growth and puberty. AB - BACKGROUND: A limited number of mutations in the GH secretagogue receptor gene (GHSR) have been described in patients with short stature. Objective To analyze GHSR in idiopathic short stature (ISS) children including a subgroup of constitutional delay of growth and puberty (CDGP) patients. SUBJECTS AND METHODS: The GHSR coding region was directly sequenced in 96 independent patients with ISS, 31 of them with CDGP, in 150 adults, and in 197 children with normal stature. The pharmacological consequences of GHSR non-synonymous variations were established using in vitro cell-based assays. RESULTS: Five different heterozygous point variations in GHSR were identified (c.-6 G>C, c.251G>T (p.Ser84Ile), c.505G>A (p.Ala169Thr), c.545 T>C (p.Val182Ala), and c.1072G>A (p.Ala358Thr)), all in patients with CDGP. Neither these allelic variants nor any other mutations were found in 694 alleles from controls. Functional studies revealed that two of these variations (p.Ser84Ile and p.Val182Ala) result in a decrease in basal activity that was in part explained by a reduction in cell surface expression. The p.Ser84Ile mutation was also associated with a defect in ghrelin potency. These mutations were identified in two female patients with CDGP (at the age of 13 years, their height SDS were -2.4 and -2.3). Both patients had normal progression of puberty and reached normal adult height (height SDS of -0.7 and -1.4) without treatment. CONCLUSION: This is the first report of GHSR mutations in patients with CDGP. Our data raise the intriguing possibility that abnormalities in ghrelin receptor function may influence the phenotype of individuals with CDGP. PMID- 21646291 TI - First impressions count: perceptions of surface-level and deep-level similarity within postnatal exercise classes and implications for program adherence. AB - The purpose of this study was to examine the extent to which perceptions of surface-level (i.e. age, ethnicity, physical condition) and deep-level (i.e. attitudes, beliefs, values) similarity, as assessed shortly after group formation, predict participants' subsequent attendance behaviors within postnatal physical activity classes. Women (N = 136) enrolled in 21 postnatal exercise classes participated in this prospective observational study. Perceptions of surface-level similarity, in particular with regard to other group members' ages, predicted women's subsequent adherence to their respective exercise classes. The results of this study suggest that perceptual similarity may have noteworthy implications for mothers' involvement within postnatal physical activity classes. PMID- 21646292 TI - Effects of adverts from a drug and alcohol prevention campaign on willingness to engage in alcohol-related risky behaviors. AB - Behavioral willingness is conceptualized as a pathway to behavior that is non deliberative, yet traditional measures require thoughtful deliberation to complete. This study explored non-deliberative measures of alcohol-related willingness to complement recent work on marijuana-related willingness. The study also examined whether adverts from a field-tested drug and alcohol prevention campaign may have operated by influencing alcohol-related willingness. Participants viewed campaign adverts or consumer adverts (control). Outcomes were reaction times to make speeded judgments about whether one would engage in risky alcohol-related behaviors. Results showed that campaign advertisements lowered willingness to play drinking games and (for males) to drive while intoxicated. PMID- 21646293 TI - Kit (W-sh) mice develop earlier and more severe experimental autoimmune encephalomyelitis due to absence of immune suppression. AB - Mast cells (MCs) have been thought to play a pathogenic role in the development of autoimmune diseases, including experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis. However, an immunoregulatory function of these cells has recently been suggested. We investigated the role of MCs in EAE using the W(-sh) mouse strain, which is MC deficient. W(-sh) mice developed earlier and more severe clinical and pathological disease with extensive demyelination and inflammation in the CNS. The inflammatory cells were mainly composed of CD4(+) T cells, monocyte/macrophages, neutrophils, and dendritic cells. Compared with wild-type mice, MC-deficient mice exhibited an increased level of MCP-1/CCR2 and CD44 expression on CD4(+) T cells in addition to decreased production of regulatory T cells, IL-4, IL-5, IL-27, and IL-10. We also found that levels of IL-17, IFN-gamma, and GM-CSF were significantly increased in peripheral lymphocytes from immunized W(-sh) mice compared with those in peripheral lymphocytes from wild-type mice. Reconstitution of W(-sh) mice downregulated susceptibility to EAE, which correlated with MC recruitment and regulatory T cell activation in the CNS. These findings indicate that responsiveness is not required in the pathogenesis of inflammatory demyelination in the CNS and that, in the absence of MCs, increased MCP-1, CCR2, IL-17, IFN gamma, CD44, and other inflammatory molecules may be responsible for increased severity of EAE. PMID- 21646294 TI - MyD88-dependent TLR1/2 signals educate dendritic cells with gut-specific imprinting properties. AB - Gut-associated dendritic cells (DC) synthesize all-trans retinoic acid, which is required for inducing gut-tropic lymphocytes. Gut-associated DC from MyD88(-/-) mice, which lack most TLR signals, expressed low levels of retinal dehydrogenases (critical enzymes for all-trans retinoic acid biosynthesis) and were significantly impaired in their ability to induce gut-homing T cells. Pretreatment of extraintestinal DC with a TLR1/2 agonist was sufficient to induce retinal dehydrogenases and to confer these DC with the capacity to induce gut homing lymphocytes via a mechanism dependent on MyD88 and JNK/MAPK. Moreover, gut associated DC from TLR2(-/-) mice, or from mice in which JNK was pharmacologically blocked, were impaired in their education to imprint gut-homing T cells, which correlated with a decreased induction of gut-tropic T cells in TLR2(-/-) mice upon immunization. Thus, MyD88-dependent TLR2 signals are necessary and sufficient to educate DC with gut-specific imprinting properties and contribute in vivo to the generation of gut-tropic T cells. PMID- 21646296 TI - FcR-independent effects of IgE and IgG autoantibodies in bullous pemphigoid. AB - Bullous pemphigoid (BP) is a subepidermal blistering disease characterized by IgE and IgG class autoantibodies specific for 180-kDa BP Ag 2 (BP180), a protein involved in cell-substrate attachment. Although some direct effects of BP IgG have been observed on keratinocytes, no study to date has examined direct effects of BP IgE. In this study, we use primary cultures of human keratinocytes to demonstrate Ag-specific binding and internalization of BP IgE. Moreover, when BP IgE and BP IgG were compared, both isotypes stimulated FcR- independent production of IL-6 and IL-8, cytokines critical for BP pathology, and elicited changes in culture confluence and viability. We then used a human skin organ culture model to test the direct effects of these Abs on the skin, whereas excluding the immune inflammatory processes that are triggered by these Abs. In these experiments, physiologic concentrations of BP IgE and BP IgG exerted similar effects on human skin by stimulating IL-6 and IL-8 production and decreasing the number of hemidesmosomes localized at the basement membrane zone. We propose that the Ab-mediated loss of hemidesmosomes could weaken attachment of basal keratinocytes to the basement membrane zone of affected skin, thereby contributing to blister formation. In this article, we identify a novel role for IgE class autoantibodies in BP mediated through an interaction with BP180 on the keratinocyte surface. In addition, we provide evidence for an FcR-independent mechanism for both IgE and IgG class autoantibodies that could contribute to BP pathogenesis. PMID- 21646295 TI - A role for p120 RasGAP in thymocyte positive selection and survival of naive T cells. AB - Activation of the Ras small GTP-binding protein is necessary for normal T cell development and function. However, it is unknown which Ras GTPase-activating proteins (RasGAPs) inactivate Ras in T cells. We used a T cell-specific RASA1 deficient mouse model to investigate the role of the p120 RasGAP (RASA1) in T cells. Death of CD4(+)CD8(+) double-positive thymocytes was increased in RASA1 deficient mice. Despite this finding, on an MHC class II-restricted TCR transgenic background, evidence was obtained for increased positive selection of thymocytes associated with augmented activation of the Ras-MAPK pathway. In the periphery, RASA1 was found to be dispensable as a regulator of Ras-MAPK activation and T cell functional responses induced by full agonist peptides. However, numbers of naive T cells were substantially reduced in RASA1-deficient mice. Loss of naive T cells in the absence of RASA1 could be attributed in part to impaired responsiveness to the IL-7 prosurvival cytokine. These findings reveal an important role for RASA1 as a regulator of double-positive survival and positive selection in the thymus as well as naive T cell survival in the periphery. PMID- 21646297 TI - The NLRP3 inflammasome is differentially activated by pneumolysin variants and contributes to host defense in pneumococcal pneumonia. AB - Streptococcus pneumoniae is a leading cause of pneumonia, meningitis, and sepsis. Pneumococci can be divided into >90 serotypes that show differences in the pathogenicity and invasiveness. We tested the hypotheses that the innate immune inflammasome pathway is involved in fighting pneumococcal pneumonia and that some invasive pneumococcal types are not recognized by this pathway. We show that human and murine mononuclear cells responded to S. pneumoniae expressing hemolytic pneumolysin by producing IL-1beta. This IL-1beta production depended on the NOD-like receptor family, pyrin domain containing 3 (NLRP3) inflammasome. Some serotype 1, serotype 8, and serotype 7F bacteria, which have previously been associated with increased invasiveness and with production of toxins with reduced hemolytic activity, or bacterial mutants lacking pneumolysin did not stimulate notable IL-1beta production. We further found that NLRP3 was beneficial for mice during pneumonia caused by pneumococci expressing hemolytic pneumolysin and was involved in cytokine production and maintenance of the pulmonary microvascular barrier. Overall, the inflammasome pathway is protective in pneumonia caused by pneumococci expressing hemolytic toxin but is not activated by clinically important pneumococcal sequence types causing invasive disease. The study indicates that a virulence factor polymorphism may substantially affect the recognition of bacteria by the innate immune system. PMID- 21646298 TI - Antibodies to the envelope glycoprotein of human T cell leukemia virus type 1 robustly activate cell-mediated cytotoxic responses and directly neutralize viral infectivity at multiple steps of the entry process. AB - Infection of human cells by human T cell leukemia virus type 1 (HTLV-1) is mediated by the viral envelope glycoproteins. The gp46 surface glycoprotein binds to cell surface receptors, including heparan sulfate proteoglycans, neuropilin 1, and glucose transporter 1, allowing the transmembrane glycoprotein to initiate fusion of the viral and cellular membranes. The envelope glycoproteins are recognized by neutralizing Abs and CTL following a protective immune response, and therefore, represent attractive components for a HTLV-1 vaccine. To begin to explore the immunological properties of potential envelope-based subunit vaccine candidates, we have used a soluble recombinant surface glycoprotein (gp46, SU) fused to the Fc region of human IgG (sRgp46-Fc) as an immunogen to vaccinate mice. The recombinant SU protein is highly immunogenic and induces high titer Ab responses, facilitating selection of hybridomas that secrete mAbs targeting SU. Many of these mAbs recognize envelope displayed on the surface of HTLV-1-infected cells and virions and several of the mAbs robustly antagonize envelope-mediated membrane fusion and neutralize pseudovirus infectivity. The most potently neutralizing mAbs recognize the N-terminal receptor-binding domain of SU, though there is considerable variation in neutralizing proficiency of the receptor binding domain-targeted mAbs. By contrast, Abs targeting the C-terminal domain of SU tend to lack robust neutralizing activity. Importantly, we find that both neutralizing and poorly neutralizing Abs strongly stimulate neutrophil-mediated cytotoxic responses to HTLV-1-infected cells. Our data demonstrate that recombinant forms of SU possess immunological features that are of significant utility to subunit vaccine design. PMID- 21646299 TI - Insulin-dependent phosphatidylinositol 3-kinase/Akt and ERK signaling pathways inhibit TLR3-mediated human bronchial epithelial cell apoptosis. AB - TLR3, one of the TLRs involved in the recognition of infectious pathogens for innate and adaptive immunity, primarily recognizes viral-associated dsRNA. Recognition of dsRNA byproducts released from apoptotic and necrotic cells is a recently proposed mechanism for the amplification of toxicity, suggesting a pivotal participation of TLR3 in viral infection, as well as in lung diseases where apoptosis plays a critical role, such as asthma and chronic obstructive pulmonary disease. In addition to metabolic control, insulin signaling was postulated to be protective by inhibiting apoptosis. Therefore, we explored the role of insulin signaling in protecting against TLR3-mediated apoptosis of human bronchial epithelial cells. Significant TLR3-mediated apoptosis was induced by polyinosinic-polycytidylic acid, a dsRNA analog, via caspase-8-dependent mechanisms. However, insulin efficiently inhibited TLR3/polyinosinic polycytidylic acid-induced human bronchial epithelial cell apoptosis via PI3K/Akt and ERK pathways, at least in part, via upregulation of cellular FLIPs and through protein synthesis-independent mechanisms. These results indicate the significance of TLR3-mediated dsRNA-induced apoptosis in the pathogenesis of apoptosis-driven lung disease and provide evidence for a novel protective role of insulin. PMID- 21646300 TI - Comment on "Acid-sensing ion channels contribute to the effect of acidosis on the function of dendritic cells". PMID- 21646301 TI - T cell lineage commitment: identity and renunciation. AB - Precursors undertaking T cell development shed their access to other pathways in a sequential process that begins before entry into the thymus and continues through many cell cycles afterward. This process involves three levels of regulatory change, in which the cells' intrinsic transcriptional regulatory factors, expression of signaling receptors (e.g., Notch1), and expression of distinct homing receptors separately contribute to confirmation of T cell identity. Each alternative potential has a different underlying molecular basis that is neutralized and then permanently silenced through different mechanisms in early T cell precursors. This regulatory mosaic has notable implications for the hierarchy of relationships linking T lymphocytes to other hematopoietic fates. PMID- 21646302 TI - Mayo Genome Consortia: a genotype-phenotype resource for genome-wide association studies with an application to the analysis of circulating bilirubin levels. AB - OBJECTIVE: To create a cohort for cost-effective genetic research, the Mayo Genome Consortia (MayoGC) has been assembled with participants from research studies across Mayo Clinic with high-throughput genetic data and electronic medical record (EMR) data for phenotype extraction. PARTICIPANTS AND METHODS: Eligible participants include those who gave general research consent in the contributing studies to share high-throughput genotyping data with other investigators. Herein, we describe the design of the MayoGC, including the current participating cohorts, expansion efforts, data processing, and study management and organization. A genome-wide association study to identify genetic variants associated with total bilirubin levels was conducted to test the genetic research capability of the MayoGC. RESULTS: Genome-wide significant results were observed on 2q37 (top single nucleotide polymorphism, rs4148325; P=5.0 * 10(-62)) and 12p12 (top single nucleotide polymorphism, rs4363657; P=5.1 * 10(-8)) corresponding to a gene cluster of uridine 5'-diphospho-glucuronosyltransferases (the UGT1A cluster) and solute carrier organic anion transporter family, member 1B1 (SLCO1B1), respectively. CONCLUSION: Genome-wide association studies have identified genetic variants associated with numerous phenotypes but have been historically limited by inadequate sample size due to costly genotyping and phenotyping. Large consortia with harmonized genotype data have been assembled to attain sufficient statistical power, but phenotyping remains a rate-limiting factor in gene discovery research efforts. The EMR consists of an abundance of phenotype data that can be extracted in a relatively quick and systematic manner. The MayoGC provides a model of a unique collaborative effort in the environment of a common EMR for the investigation of genetic determinants of diseases. PMID- 21646303 TI - Deep brain stimulation: current and future clinical applications. AB - Deep brain stimulation (DBS) has developed during the past 20 years as a remarkable treatment option for several different disorders. Advances in technology and surgical techniques have essentially replaced ablative procedures for most of these conditions. Stimulation of the ventralis intermedius nucleus of the thalamus has clearly been shown to markedly improve tremor control in patients with essential tremor and tremor related to Parkinson disease. Symptoms of bradykinesia, tremor, gait disturbance, and rigidity can be significantly improved in patients with Parkinson disease. Because of these improvements, a decrease in medication can be instrumental in reducing the disabling features of dyskinesias in such patients. Primary dystonia has been shown to respond well to DBS of the globus pallidus internus. The success of these procedures has led to application of these techniques to multiple other debilitating conditions such as neuropsychiatric disorders, intractable pain, epilepsy, camptocormia, headache, restless legs syndrome, and Alzheimer disease. The literature analysis was performed using a MEDLINE search from 1980 through 2010 with the term deep brain stimulation, and several double-blind and larger case series were chosen for inclusion in this review. The exact mechanism of DBS is not fully understood. This review summarizes many of the current and potential future clinical applications of this technology. PMID- 21646304 TI - Evidence that venous hypertension causes stasis dermatitis. AB - A clinical model to examine the hypothesis that venous hypertension of the lower leg per se can cause lower leg stasis dermatitis is described. To prove this concept, we retrospectively studied a consecutive series of 38 patients with lower leg dermatitis who underwent phlebological examination at our consultation over a period of four years. Among those patients who had an insufficiency of the superficial veins only, without insufficiency of the deep veins, 22 had undergone patch testing to common allergens in phlebology. We found 10 patients with a stasis dermatitis of the lower leg and an incompetent great saphenous vein, six of whom had no detectable contact sensitization at all and another four exclusively to phlebologically irrelevant substances, e.g. nickel, cobalt, chromate or epoxid resin. All these 10 patients showed long saphenous vein incompetence from the groin to the medial aspect of the leg. All were operated by classical flush ligation and saphenectomy. Lower leg dermatitis healed in all 10 patients within 8-12 weeks and no recurrence was observed (1 year follow-up). These results support clinical experience that venous hypertension alone indeed can cause lower leg dermatitis. PMID- 21646305 TI - Cortactin and focal adhesion kinase as predictors of cancer risk in patients with laryngeal premalignancy. AB - Novel markers are needed to accurately predict the risk of malignant transformation in laryngeal premalignancies. We therefore investigated the clinical significance of cortactin (CTTN) and focal adhesion kinase (FAK) during laryngeal tumorigenesis and their potential utility as cancer risk markers. CTTN and FAK protein expression and gene amplification were assessed in 82 patients with laryngeal dysplasia and correlated with clinicopathologic parameters and laryngeal cancer risk. Increased CTTN and FAK expression was found respectively in 41 (50%) and 40 (49%) of 82 laryngeal dysplasias; protein expression was maintained or further augmented in the corresponding patient-matched invasive tumors subsequently developed. CTTN and FAK/PTK2 gene amplifications were respectively detected in 10 (12%) and 26 (32%) laryngeal dysplasias. Both CTTN and FAK protein expression increased with the grade of dysplasia; however, CTTN and FAK expression but not histology correlated significantly with increased laryngeal cancer risk (P = 0.009 and P = 0.002, respectively). Patients carrying strong CTTN- or FAK-expressing dysplastic lesions experienced a significantly higher cancer incidence (P = 0.006 and P = 0.001, respectively; log-rank test). Furthermore, FAK expression was an independent predictor of laryngeal cancer development (HR = 3.706, 95% CI: 1.735-7.916; P = 0.001) and the combination of FAK and CTTN showed superior predictive value (HR = 5.042, 95% CI: 2.255-11.274; P < 0.001). Taken together, our findings support the involvement of CTTN and FAK in malignant transformation and provide original evidence for their potential clinical utility as biomarkers for the risk of developing laryngeal cancer. PMID- 21646306 TI - Unwilling executioners? PMID- 21646307 TI - Science asks researchers to withdraw paper on chronic fatigue syndrome and infectious retrovirus. PMID- 21646308 TI - Palliative care is "neglected" worldwide, report says. PMID- 21646309 TI - NHS trust admits negligent incontinence care harmed many women. PMID- 21646310 TI - Global leaders call for end to war on drugs. PMID- 21646311 TI - A record 6.6 million patients in poor countries received antiretroviral treatment in 2010. PMID- 21646312 TI - London vaccination summit offers "unprecedented opportunity to save millions of lives". PMID- 21646313 TI - Liverpool University reinstates course on social exclusion after protests. PMID- 21646314 TI - Subclinical hypothyroidism and its association with lupus nephritis: a case control study in a large cohort of Chinese systemic lupus erythematosus patients. AB - The aim of this study was to evaluate the prevalence of thyroid diseases in Chinese systemic lupus erythematosus (SLE) patients and the relevance of subclinical hypothyroidism (SCH) with lupus nephritis (LN). A large cohort of 1006 SLE patients was retrospectively analyzed. The prevalence of autoimmune thyroid disease was 2.78%, clinical hypothyroidism 1.69%, subclinical hypothyroidism 10.04%, central hypothyroidism 1.29%, hyperthyroidism 1.19%, euthyroid sick syndrome (ESS) 9.54%, and nodules 1.09%, respectively. Compared with the prevalence of thyroid abnormalities in the general Chinese population (0.91-6.05%), SCH was much higher (10.04%) in this study. In addition, SCH was more frequent in patients with LN (13.4%) than those without LN (7.3%, p = 0.001). Case control study was performed to explore the relative risk factors of SCH. In multiple logistic regression models, 24 h urine protein and estimated glomerular filtration rate (eGFR) were retained as independent correlates of SCH after adjusting for demographic variables, risk factors, and other potential confounders. The results of the present study suggest that SCH is a common complication in SLE patients, and closely related with LN. PMID- 21646315 TI - Vitamin D insufficiency in Brazilian patients with SLE: prevalence, associated factors, and relationship with activity. AB - The influence of vitamin D on the severity of systemic lupus erythematosus (SLE) has been studied. Our objectives were to determine the prevalence of vitamin D insufficiency in Brazilian lupus patients and its relationship with demographic, clinical, and laboratory variables and to study the relationship between vitamin D insufficiency and disease activity. This is a cross-sectional study of 159 SLE patients. Levels of serum 25-hydroxyvitamin D (25(OH)D) <30 ng/ml were defined as vitamin D insufficiency and <20 ng/ml as deficiency. Data collected included sex, age, ethnicity, postmenopausal status, disease duration, cumulative amount of oral glucocorticoids (GC) in the last six months, duration of GC usage, calcium and vitamin D supplements, photosensitivity, sunscreen usage, sun exposure, diabetes mellitus, creatinine clearance, lifestyle habits, and smoking. Disease activity was measured by the SLE Disease Activity Index 2000 (SLEDAI-2K). Bivariate statistical analysis was performed using the Mann-Whitney test, Spearman's correlation coefficient, Chi-square test, and Fisher's exact test. Multivariate analysis was performed with multiple linear regression. The prevalence of vitamin D insufficiency and deficiency were 37.7% and 8.2%, respectively. Levels of 25(OH) D were not associated with lupus activity score, disease duration, sun exposure, vitamin D supplementation, or use of corticosteroids. PMID- 21646316 TI - Valsartan is more effective than placebo in reducing the incidence of diabetes in people with impaired glucose tolerance and cardiovascular disease or risk factors but has no effect on cardiovascular outcomes. PMID- 21646317 TI - People aged above 65 treated for latent tuberculosis are at increased risk of admission for a hepatic event compared with untreated controls. PMID- 21646320 TI - Barriers and facilitators to consulting hospital clinical ethics committees. AB - Hospitals in many countries have had clinical ethics committees for over 20 years. Despite this, there has been little research to evaluate these committees and growing evidence that they are underutilized. To address this gap, we investigated the question 'What are the barriers and facilitators nurses and physicians perceive in consulting their hospital ethics committee?' Thirty-four nurses, 10 nurse managers and 31 physicians working at four Canadian hospitals were interviewed using a semi-structured interview guide as part of a larger investigation. We used content analysis of the interview data related to barriers and facilitators to use of hospital ethics committees to identify nine categories of barriers and nine categories of facilitators. These categories as well as their subcategories are discussed and those specific to nurses or physicians are identified. The need to increase health professionals' use of clinical ethics committees through reducing barriers and maximizing facilitators is discussed. PMID- 21646321 TI - Engaging with children in research: Theoretical and practical implications of negotiating informed consent/assent. AB - At the outset of an ethnographic inquiry, we navigated national and international resources to search for theoretical and practical guidance on obtaining parents and children's informed consent/assent. While much theoretical guidance debating ethical issues to children's participation in research was found, a paucity of published papers offering practical guidance on assent processes and/or visual representations of child assent forms and information sheets was discovered. The purpose of this article is to describe our experiences, both theoretically and practically, of negotiating the process of obtaining informed consent and assent with parents and children for a non-therapeutic qualitative research study. We hope this article instigates a platform for others to explicate their experience and contributes to the construction of a coherent body of knowledge which will clearly delineate important elements that must be considered when negotiating children's agreement to participate in research. PMID- 21646322 TI - The ethical component of professional competence in nursing: an analysis. AB - The purpose of this article is to initiate a philosophical discussion about the ethical component of professional competence in nursing from the perspective of Brazilian nurses. Specifically, this article discusses professional competence in nursing practice in the Brazilian health context, based on two different conceptual frameworks. The first framework is derived from the idealistic and traditional approach while the second views professional competence through the lens of historical and dialectical materialism theory. The philosophical analyses show that the idealistic view of professional competence differs greatly from practice. Combining nursing professional competence with philosophical perspectives becomes a challenge when ideals are opposed by the reality and implications of everyday nursing practice. PMID- 21646323 TI - Nursing strikes: an ethical perspective on the US healthcare community. AB - Recent labor disputes between registered nurses and hospitals in Minnesota, California, and Pennsylvania raise moral questions about nurses' professional obligations, nurses' right to collectively bargain to preserve or improve wages, benefits, and working conditions, and patients' right to medical care. Deontology and consequentialism focus too narrowly on nurses and patients, and thus ignore the nature of the healthcare community as a system of competing interests. When considered in this context, nurses' strikes are shown to be consistent with this system of competing interests, and thus are morally permissible. PMID- 21646324 TI - Nursing under the influence: a relational ethics perspective. AB - When nurses have active and untreated addictions, patient safety may be compromised and nurse-health endangered. Genuine responses are required to fulfil nurses' moral obligations to their patients as well as to their nurse-colleagues. Guided by core elements of relational ethics, the influences of nursing organizational responses along with the practice environment in shaping the situation are contemplated. This approach identifies the importance of consistency with nursing values, acknowledges nurses interdependence, and addresses the role of nursing organization as moral agent. By examining the relational space, the tension between what appears to be opposing moral responsibilities may be healed. Ongoing discourse to identify authentic actions for the professional practice issue of nursing under the influence is called upon. PMID- 21646325 TI - Factors influencing preferences of Korean people toward advance directives. AB - Although Korean society has begun to seek a way of utilizing advance directives, there is not much known about the factors influencing the average Korean person's preference toward advance directives. The purpose of this study was to examine factors, in addition to demographic variables, influencing preferences regarding advance directives. These include: to what extent people's awareness of advance directives, preferences of extending their life at the end of life, experience of illness and medical care, and family functioning independently influence the preferences toward advance directives. The participants were 382 community dwelling Korean people. The data analysis was performed using hierarchical multiple logistic regression analysis. The findings showed that a majority of Korean people had a positive preference on advance directives and the factors influencing their preferences for advance directives were the preferences against the use of life-sustaining treatment at the end of life, a good self-rated heath status, and an unsatisfactory family functioning. PMID- 21646326 TI - Academic dishonesty among nursing students: a descriptive study. AB - This descriptive and cross-sectional study aims to evaluate academic dishonesty among university nursing students in Turkey. The study's sample included 196 students. Two instruments were used for gathering data. The first instrument, a questionnaire, which included some socio-demographic variables (age, class, gender, education, family structure, parents' attitude and educators' attitude) formed the first part. The second part included the Academic Dishonesty Tendency Scale developed by Eminoglu and Nartgun. The data were analyzed using descriptive statistics and Kruskall Wallis, One-way Anova, t- test and Mann-Whitney U test. It was found that academic dishonesty was at medium-level (2.60-3.39) in nursing students. PMID- 21646327 TI - Protecting vulnerable research participants: a Foucault-inspired analysis of ethics committees. AB - History has demonstrated the necessity of protecting research participants. Research ethics are based on a concept of asymmetry of power, viewing the researcher as powerful and potentially dangerous and establishing ethics committees as external agencies in the field of research. We argue in favour of expanding this perspective on relationships of power to encompass the ethics committees as one among several actors that exert power and that act in a relational interplay with researchers and participants. We employ Michel Foucault's ideas of power as an omnipresent force which is dynamic and unstable, as well as the notion that knowledge and power are inextricably intertwined. The article discusses how research ethics committees may affect academic freedom. In addition it is pointed out that research participants could be harmed - not only by unfortunate research practices, but also by being subjected to the protective efforts of ethics monitoring bodies. PMID- 21646328 TI - Paediatrics: the etymology of a name. AB - Within the history of paediatrics is the history of the name used to describe it. The etymology of the word 'paediatrics' dates from its first written use, recorded as 'padiatrik' in the German literature and as 'paediatric', later 'pediatric' in the USA, both first in 1850. Professor Robley Dunglison (1788 1869), the British and American medical lexicographer, first defined 'paediatria' as 'the treatment of the diseases of children' in 1855. 'Pediatric medicine' was promoted as a specialty in the USA in 1880. The oldest monumental inscription defining the specialty of 'paediatrics' in the UK is to be found on a plaque added (in 1950) to the memorial to Dr George Armstrong (1719-1789), a founder of the specialty of paediatrics, in Castleton Cemetery, Scottish Borders, Roxburghshire. 'Paediatrics' and 'child health', with subtle semantic distinctions, had become well established in the English-speaking world by the middle of the 20th century. This paper presents an interpretative chronology of the etymology of the descriptors of the specialty that enjoins all who care for children. PMID- 21646329 TI - Unilateral subdural motor cortex stimulation improves essential tremor but not Parkinson's disease. AB - Epidural motor cortex stimulation has been reported to be effective in treating some movement disorders. Nevertheless, clinical results have been variable and no double-blinded evaluations have been reported. The aim of this study was to investigate efficacy and safety of unilateral subdural motor cortex stimulation in patients with essential tremor and Parkinson's disease. Six patients with essential tremor and five parkinsonian patients were selected. Craniotomy was performed under local anaesthesia with conscious sedation. A four contact electrode (Resume II model 3587, Medtronic, Inc) was positioned on the motor cortex, after identification of the area with direct monopolar cortical stimulation. Soon after surgery, a variety of different settings of stimulation were assessed using standard rating scales to select the optimal stimulation parameters. The effects of chronic stimulation were evaluated in both groups of patients after 3 months (double-blinded fashion) and 1 year (open fashion). In essential tremor, contralateral hand tremor scores significantly improved (P = 0.04) with stimulation during the double-blinded study, whereas in Parkinson's disease, there were no changes in the OFF medication/on stimulation motor scores compared with off stimulation. At 1 year, tremor was improved by stimulation in two out of three patients with essential tremor available at follow-up, whereas no improvement was observed in the five parkinsonian patients. One parkinsonian patient had a cortical venous infarct. Three other patients had self-limiting seizures with aggressive trials of stimulation in the period of dosage selection. These findings suggest that unilateral subdural motor cortex stimulation may be useful for contralateral hand tremor in selected patients with essential tremor but was not effective in improving parkinsonian signs in our series. PMID- 21646330 TI - Spastic paraplegia in 'dominant optic atrophy plus' phenotype due to OPA1 mutation. PMID- 21646331 TI - Atrophy, hypometabolism and white matter abnormalities in semantic dementia tell a coherent story. AB - Semantic dementia, in which there is progressive deterioration of semantic knowledge, is associated with focal, typically asymmetric, temporal lobe degeneration. The ventrorostral temporal lobe is most severely affected and there is concordance between atrophy and reduced metabolic activity. In this study, we confirmed the veracity of this claim using 18F-fluorodeoxyglucose positron emission tomography and anatomical magnetic resonance images. The principal aim, however, was to understand the impact on neuronal projections from the ventrorostral temporal cortex lesion by studying the full extent of white matter changes, with no a priori assumptions about the nature or spatial location of the tracts involved. Using an unbiased voxel-wise approach known as tract-based spatial statistics, we compared results of whole-brain diffusion tensor imaging- absolute metrics of axial, radial and mean diffusion as well as fractional anisotropy--from 10 patients with mild/moderate semantic dementia and 21 matched controls. Distributions of increased absolute diffusivity and reduced fractional anisotropy for patients with semantic dementia were spatially concordant with each other. Abnormalities in all metrics were highly statistically significant in ventrorostral temporal white matter, more extreme on the left side, thus closely matching results from structural and functional imaging of grey matter. The most sensitive marker of change was radial diffusion. Local white matter tract abnormalities extended rostrally towards the frontal lobe and dorsocaudally towards the superior temporal and supramarginal gyri. To examine more remote changes, we performed a skeletonized probabilistic tractography analysis- 'seeding' the rostral temporal voxels identified as abnormal in the patient group -in a healthy control group. Three major neural pathways were found to emanate from this 'seed region': uncinate, arcuate and inferior longitudinal fasciculi. At a less conservative threshold, tensor abnormalities in the semantic dementia group mapped onto the tractographies for the uncinate and arcuate bundles well beyond the rostral temporal lobe; this was not the case for the inferior longitudinal bundle, where abnormalities in semantic dementia did not extend caudal to the atrophic/hypometabolic zone. The results offer direct evidence for how the ventrorostral temporal lesion, proposed to be responsible for deteriorating semantic knowledge in semantic dementia and separate from 'classic' language areas, is associated with degeneration of efferent white matter projections to such language areas. PMID- 21646332 TI - Multiple imputation models should incorporate the outcome in the model of interest. PMID- 21646333 TI - Differentially regulated splice variants and systems biology analysis of Kaposi's sarcoma-associated herpesvirus-infected lymphatic endothelial cells. AB - Alternative RNA splicing greatly increases proteome diversity, and the possibility of studying genome-wide alternative splicing (AS) events becomes available with the advent of high-throughput genomics tools devoted to this issue. Kaposi's sarcoma associated herpesvirus (KSHV) is the etiological agent of KS, a tumor of lymphatic endothelial cell (LEC) lineage, but little is known about the AS variations induced by KSHV. We analyzed KSHV-controlled AS using high-density microarrays capable of detecting all exons in the human genome. Splicing variants and altered exon-intron usage in infected LEC were found, and these correlated with protein domain modification. The different 3'-UTR used in new transcripts also help isoforms to escape microRNA-mediated surveillance. Exome-level analysis further revealed information that cannot be disclosed using classical gene-level profiling: a significant exon usage difference existed between LEC and CD34(+) precursor cells, and KSHV infection resulted in LEC-to precursor, dedifferentiation-like exon level reprogramming. Our results demonstrate the application of exon arrays in systems biology research, and suggest the regulatory effects of AS in endothelial cells are far more complex than previously observed. This extra layer of molecular diversity helps to account for various aspects of endothelial biology, KSHV life cycle and disease pathogenesis that until now have been unexplored. PMID- 21646334 TI - A multifunctional bioconjugate module for versatile photoaffinity labeling and click chemistry of RNA. AB - A multifunctional reagent based on a coumarin scaffold was developed for derivatization of naive RNA. The alkylating agent N3BC [7-azido-4 (bromomethyl)coumarin], obtained by Pechmann condensation, is selective for uridine. N3BC and its RNA conjugates are pre-fluorophores which permits controlled modular and stepwise RNA derivatization. The success of RNA alkylation by N3BC can be monitored by photolysis of the azido moiety, which generates a coumarin fluorophore that can be excited with UV light of 320 nm. The azidocoumarin-modified RNA can be flexibly employed in structure-function studies. Versatile applications include direct use in photo-crosslinking studies to cognate proteins, as demonstrated with tRNA and RNA fragments from the MS2 phage and the HIV genome. Alternatively, the azide function can be used for further derivatization by click-chemistry. This allows e.g. the introduction of an additional fluorophore for excitation with visible light. PMID- 21646335 TI - The Gene3D Web Services: a platform for identifying, annotating and comparing structural domains in protein sequences. AB - The Gene3D structural domain database provides domain annotations for 7 million proteins, based on the manually curated structural domain superfamilies in CATH. These annotations are integrated with functional, genomic and molecular information from external resources, such as GO, EC, UniProt and the NCBI Taxonomy database. We have constructed a set of web services that provide programmatic access to this integrated database, as well as the Gene3D domain recognition tool (Gene3DScan) and protein sequence annotation pipeline for analysing novel protein sequences. Example queries include retrieving all curated GO terms for a domain superfamily or all the multi-domain architectures for the human genome. The services can be accessed using simple HTTP calls and are able to return results in a range of formats for quick downloading and easy parsing, graphical rendering and data storage. Hence, they provide a simple, but flexible means of integrating domain annotations and associated data sets into locally run pipelines and analysis software. The services can be found at http://gene3d.biochem.ucl.ac.uk/WebServices/. PMID- 21646336 TI - Phylemon 2.0: a suite of web-tools for molecular evolution, phylogenetics, phylogenomics and hypotheses testing. AB - Phylemon 2.0 is a new release of the suite of web tools for molecular evolution, phylogenetics, phylogenomics and hypotheses testing. It has been designed as a response to the increasing demand of molecular sequence analyses for experts and non-expert users. Phylemon 2.0 has several unique features that differentiates it from other similar web resources: (i) it offers an integrated environment that enables evolutionary analyses, format conversion, file storage and edition of results; (ii) it suggests further analyses, thereby guiding the users through the web server; and (iii) it allows users to design and save phylogenetic pipelines to be used over multiple genes (phylogenomics). Altogether, Phylemon 2.0 integrates a suite of 30 tools covering sequence alignment reconstruction and trimming; tree reconstruction, visualization and manipulation; and evolutionary hypotheses testing. PMID- 21646337 TI - In vivo functional analysis of the Dicistroviridae intergenic region internal ribosome entry sites. AB - Some viral and cellular messages use an alternative mechanism to initiate protein synthesis that involves internal recruitment of the ribosome to an internal ribosome entry site (IRES). The Dicistroviridae intergenic regions (IGR) have been studied as model IRESs to understand the mechanism of IRES-mediated translation. In this study, the in vivo activity of IGR IRESs were compared. Our analysis demonstrates that Class I and II IGR IRESs have comparable translation efficiency in yeast and that Class II is significantly more active in mammalian cells. Furthermore, while Class II IGR IRES activity was enhanced in yeast grown at a higher temperature, temperature did not affect IGR IRES activity in mammalian cells. This suggests that Class II IRESs may not function optimally with yeast ribosomes. Examination of chimeric IGR IRESs, established that the IRES strength and temperature sensitivity are mediated by the ribosome binding domain. In addition, the sequence of the first translated codon is also an important determinant of IRES activity. Our findings provide us with a comprehensive overview of IGR IRES activities and allow us to begin to understand the differences between Classes I and II IGR IRESs. PMID- 21646338 TI - Homogeneous antibody-based proximity extension assays provide sensitive and specific detection of low-abundant proteins in human blood. AB - Convenient and well-performing protein detection methods for a wide range of targets are in great demand for biomedical research and future diagnostics. Assays without the need for washing steps while still unaffected when analyzing complex biological samples are difficult to develop. Herein, we report a well characterized nucleic acid proximity-based assay using antibodies, called Proximity Extension Assay (PEA), showing good performance in plasma samples. Target-specific antibody pairs are linked to DNA strands that upon simultaneous binding to the target analyte create a real-time PCR amplicon in a proximity dependent manner enabled by the action of a DNA polymerase. 3'Exonuclease-capable polymerases were found to be clearly superior in sensitivity over non 3'exonuclease ones. A PEA was set up for IL-8 and GDNF in a user-friendly, homogenous assay displaying femtomolar detection sensitivity, good recovery in human plasma, high specificity and up to 5-log dynamic range in 1 MUL samples. Furthermore, we have illustrated the use of a macro-molecular crowding matrix in combination with this homogeneous assay to drive target binding for low-affinity antibodies, thereby improving the sensitivity and increasing affinity reagent availability by lowering assay development dependency on high-affinity antibodies. Assay performance was also confirmed for a multiplex version of PEA. PMID- 21646339 TI - PileLineGUI: a desktop environment for handling genome position files in next generation sequencing studies. AB - Next-generation sequencing (NGS) technologies are making sequence data available on an unprecedented scale. In this context, new catalogs of Single Nucleotide Polymorphism and mutations generated by resequencing studies are usually stored in genome position files (e.g. Variant Call Format, SAMTools pileup, BED, GFF) comprising of large lists of genomic positions, which are difficult to handle by researchers. Here, we present PileLineGUI, a novel desktop application primarily designed for manipulating, browsing and analysing genome position files (GPF), with specific support to somatic mutation finding studies. The developed tool also integrates a new genome browser module specially designed for inspecting GPFs. PileLineGUI is free, multiplatform and designed to be intuitively used by biomedical researchers. PileLineGUI is available at: http://sing.ei.uvigo.es/pileline/pilelinegui.html. PMID- 21646340 TI - UV stalled replication forks restart by re-priming in human fibroblasts. AB - Restarting stalled replication forks is vital to avoid fatal replication errors. Previously, it was demonstrated that hydroxyurea-stalled replication forks rescue replication either by an active restart mechanism or by new origin firing. To our surprise, using the DNA fibre assay, we only detect a slightly reduced fork speed on a UV-damaged template during the first hour after UV exposure, and no evidence for persistent replication fork arrest. Interestingly, no evidence for persistent UV-induced fork stalling was observed even in translesion synthesis defective, Poleta(mut) cells. In contrast, using an assay to measure DNA molecule elongation at the fork, we observe that continuous DNA elongation is severely blocked by UV irradiation, particularly in UV-damaged Poleta(mut) cells. In conclusion, our data suggest that UV-blocked replication forks restart effectively through re priming past the lesion, leaving only a small gap opposite the lesion. This allows continuation of replication on damaged DNA. If left unfilled, the gaps may collapse into DNA double-strand breaks that are repaired by a recombination pathway, similar to the fate of replication forks collapsed after hydroxyurea treatment. PMID- 21646341 TI - An alternative approach to multiple genome comparison. AB - Genome comparison is now a crucial step for genome annotation and identification of regulatory motifs. Genome comparison aims for instance at finding genomic regions either specific to or in one-to-one correspondence between individuals/strains/species. It serves e.g. to pre-annotate a new genome by automatically transferring annotations from a known one. However, efficiency, flexibility and objectives of current methods do not suit the whole spectrum of applications, genome sizes and organizations. Innovative approaches are still needed. Hence, we propose an alternative way of comparing multiple genomes based on segmentation by similarity. In this framework, rather than being formulated as a complex optimization problem, genome comparison is seen as a segmentation question for which a single optimal solution can be found in almost linear time. We apply our method to analyse three strains of a virulent pathogenic bacteria, Ehrlichia ruminantium, and identify 92 new genes. We also find out that a substantial number of genes thought to be strain specific have potential orthologs in the other strains. Our solution is implemented in an efficient program, qod, equipped with a user-friendly interface, and enables the automatic transfer of annotations between compared genomes or contigs (Video in Supplementary Data). Because it somehow disregards the relative order of genomic blocks, qod can handle unfinished genomes, which due to the difficulty of sequencing completion may become an interesting characteristic for the future. Availabilty: http://www.atgc-montpellier.fr/qod. PMID- 21646342 TI - CSpritz: accurate prediction of protein disorder segments with annotation for homology, secondary structure and linear motifs. AB - CSpritz is a web server for the prediction of intrinsic protein disorder. It is a combination of previous Spritz with two novel orthogonal systems developed by our group (Punch and ESpritz). Punch is based on sequence and structural templates trained with support vector machines. ESpritz is an efficient single sequence method based on bidirectional recursive neural networks. Spritz was extended to filter predictions based on structural homologues. After extensive testing, predictions are combined by averaging their probabilities. The CSpritz website can elaborate single or multiple predictions for either short or long disorder. The server provides a global output page, for download and simultaneous statistics of all predictions. Links are provided to each individual protein where the amino acid sequence and disorder prediction are displayed along with statistics for the individual protein. As a novel feature, CSpritz provides information about structural homologues as well as secondary structure and short functional linear motifs in each disordered segment. Benchmarking was performed on the very recent CASP9 data, where CSpritz would have ranked consistently well with a Sw measure of 49.27 and AUC of 0.828. The server, together with help and methods pages including examples, are freely available at URL: http://protein.bio.unipd.it/cspritz/. PMID- 21646343 TI - g:Profiler--a web server for functional interpretation of gene lists (2011 update). AB - Functional interpretation of candidate gene lists is an essential task in modern biomedical research. Here, we present the 2011 update of g:Profiler (http://biit.cs.ut.ee/gprofiler/), a popular collection of web tools for functional analysis. g:GOSt and g:Cocoa combine comprehensive methods for interpreting gene lists, ordered lists and list collections in the context of biomedical ontologies, pathways, transcription factor and microRNA regulatory motifs and protein-protein interactions. Additional tools, namely the biomolecule ID mapping service (g:Convert), gene expression similarity searcher (g:Sorter) and gene homology searcher (g:Orth) provide numerous ways for further analysis and interpretation. In this update, we have implemented several features of interest to the community: (i) functional analysis of single nucleotide polymorphisms and other DNA polymorphisms is supported by chromosomal queries; (ii) network analysis identifies enriched protein-protein interaction modules in gene lists; (iii) functional analysis covers human disease genes; and (iv) improved statistics and filtering provide more concise results. g:Profiler is a regularly updated resource that is available for a wide range of species, including mammals, plants, fungi and insects. PMID- 21646344 TI - Systematic bias in high-throughput sequencing data and its correction by BEADS. AB - Genomic sequences obtained through high-throughput sequencing are not uniformly distributed across the genome. For example, sequencing data of total genomic DNA show significant, yet unexpected enrichments on promoters and exons. This systematic bias is a particular problem for techniques such as chromatin immunoprecipitation, where the signal for a target factor is plotted across genomic features. We have focused on data obtained from Illumina's Genome Analyser platform, where at least three factors contribute to sequence bias: GC content, mappability of sequencing reads, and regional biases that might be generated by local structure. We show that relying on input control as a normalizer is not generally appropriate due to sample to sample variation in bias. To correct sequence bias, we present BEADS (bias elimination algorithm for deep sequencing), a simple three-step normalization scheme that successfully unmasks real binding patterns in ChIP-seq data. We suggest that this procedure be done routinely prior to data interpretation and downstream analyses. PMID- 21646345 TI - Phosphorylation of histone H2B serine 32 is linked to cell transformation. AB - Various types of post-translational modifications of the histone tails have been revealed, but a few modifications have been found within the histone core sequences. Histone core post-translational modifications have the potential to modulate nucleosome structure and DNA accessibility. Here, we studied the histone H2B core domain and found that phosphorylation of H2B serine 32 occurs in normal cycling and mitogen-stimulated cells. Notably, this phosphorylation is elevated in skin cancer cell lines and tissues compared with normal counterparts. The JB6 Cl41 mouse skin epidermal cell line is a well established model for tumor promoter-induced cell transformation and was used to study the function of H2B during EGF-induced carcinogenesis. Remarkably, cells overexpressing a nonphosphorylatable H2BS32A mutant exhibited suppressed growth and EGF-induced cell transformation, possibly because of decreased activation of activator protein-1, compared with control cells overexpressing wild type H2B. We identified ribosomal S6 kinase 2 (RSK2) as the kinase responsible for H2BS32 phosphorylation. Serum-starved JB6 cells contain very little endogenous H2BS32 phosphorylation, and EGF treatment induced this phosphorylation. The phosphorylation was attenuated in RSK2 knock-out MEFs and RSK2 knockdown JB6 cells. Taken together, our results demonstrate a novel role for H2B phosphorylation in cell transformation and show that H2BS32 phosphorylation is critical for controlling activator protein-1 activity, which is a major driver in cell transformation. PMID- 21646346 TI - MicroRNA expression profiles of human blood monocyte-derived dendritic cells and macrophages reveal miR-511 as putative positive regulator of Toll-like receptor 4. AB - Dendritic cells (DCs) and macrophages (MFs) are important multifunctional immune cells. Like other cell types, they express hundreds of different microRNAs (miRNAs) that are recently discovered post-transcriptional regulators of gene expression. Here we present updated miRNA expression profiles of monocytes, DCs and MFs. Compared with monocytes, ~50 miRNAs were found to be differentially expressed in immature and mature DCs or MFs, with major expression changes occurring during the differentiation. Knockdown of DICER1, a protein needed for miRNA biosynthesis, led to lower DC-specific intercellular adhesion molecule-3 grabbing non-integrin (DC-SIGN) and enhanced CD14 protein levels, confirming the importance of miRNAs in DC differentiation in general. Inhibition of the two most highly up-regulated miRNAs, miR-511 and miR-99b, also resulted in reduced DC-SIGN level. Prediction of miRNA-511 targets revealed a number of genes with known immune functions, of which TLR4 and CD80 were validated using inhibition of miR 511 in DCs and luciferase assays in HEK293 cells. Interestingly, under the cell cycle arrest conditions, miR-511 seems to function as a positive regulator of TLR4. In conclusion, we have identified miR-511 as a novel potent modulator of human immune response. In addition, our data highlight that miRNA influence on gene expression is dependent on the cellular environment. PMID- 21646347 TI - Identification of a replication-independent replacement histone H3 in the basidiomycete Ustilago maydis. AB - Ustilago maydis is a haploid basidiomycete with single genes for two distinct histone H3 variants. The solitary U1 gene codes for H3.1, predicted to be a replication-independent replacement histone. The U2 gene is paired with histone H4 and produces a putative replication-coupled H3.2 variant. These predictions were evaluated experimentally. U2 was confirmed to be highly expressed in the S phase and had reduced expression in hydroxyurea, and H3.2 protein was not incorporated into transcribed chromatin of stationary phase cells. Constitutive expression of U1 during growth produced ~25% of H3 as H3.1 protein, more highly acetylated than H3.2. The level of H3.1 increased when cell proliferation slowed, a hallmark of replacement histones. Half of new H3.1 incorporated into highly acetylated chromatin was lost with a half-life of 2.5 h, the fastest rate of replacement H3 turnover reported to date. This response reflects the characteristic incorporation of replacement H3 into transcribed chromatin, subject to continued nucleosome displacement and a loss of H3 as in animals and plants. Although the two H3 variants are functionally distinct, neither appears to be essential for vegetative growth. KO gene disruption transformants of the U1 and U2 loci produced viable cell lines. The structural and functional similarities of the Ustilago replication-coupled and replication-independent H3 variants with those in animals, in plants, and in ciliates are remarkable because these distinct histone H3 pairs of variants arose independently in each of these clades and in basidiomycetes. PMID- 21646348 TI - Reactive oxygen species mediate bactericidal killing elicited by carbon monoxide releasing molecules. AB - CO-releasing molecules (CO-RMs) were previously shown by us to be more potent bactericides than CO gas. This suggests a mechanism of action for CO-RM, which either potentiates the activity of CO or uses another CO-RM-specific effect. We have also reported that CORM-2 induces the expression of genes related to oxidative stress. In the present study we intend to establish whether the generation of reactive oxygen species by CO-RMs may indeed result in the inhibition of bacterial cellular function. We now report that two CO-RMs (CORM-2 and ALF062) stimulate the production of ROS in Escherichia coli, an effect that is abolished by addition of antioxidants. Furthermore, deletion of genes encoding E. coli systems involved in reactive oxygen species scavenging, namely catalases and superoxide dismutases, potentiates the lethality of CORM-2 due to an increase of intracellular ROS content. CORM-2 also induces the expression of the E. coli DNA repair/SOS system recA, and its inactivation enhances toxicity of CORM-2. Moreover, fluorescence microscopy images reveal that CORM-2 causes DNA lesions to bacterial cells. We also demonstrate that cells treated with CORM-2 contain higher levels of free iron arising from destruction of iron-sulfur proteins. Importantly, we show that CO-RMs generate hydroxyl radicals in a cell-free solution, a process that is abolished by scavenging CO. Altogether, we provide a novel insight into the molecular basis of CO-RMs action by showing that their bactericidal properties are linked to cell damage inflicted by the oxidative stress that they are able to generate. PMID- 21646349 TI - Tumor necrosis factor inhibits glucocorticoid receptor function in mice: a strong signal toward lethal shock. AB - As glucocorticoid resistance (GCR) and the concomitant burden pose a worldwide problem, there is an urgent need for a more effective glucocorticoid therapy, for which insights into the molecular mechanisms of GCR are essential. In this study, we addressed the hypothesis that TNFalpha, a strong pro-inflammatory mediator in numerous inflammatory diseases, compromises the protective function of the glucocorticoid receptor (GR) against TNFalpha-induced lethal inflammation. Indeed, protection of mice by dexamethasone against TNFalpha lethality was completely abolished when it was administered after TNFalpha stimulation, indicating compromised GR function upon TNFalpha challenge. TNFalpha-induced GCR was further demonstrated by impaired GR-dependent gene expression in the liver. Furthermore, TNFalpha down-regulates the levels of both GR mRNA and protein. However, this down-regulation seems to occur independently of GC production, as TNFalpha also resulted in down-regulation of GR levels in adrenalectomized mice. These findings suggest that the decreased amount of GR determines the GR response and outcome of TNFalpha-induced shock, as supported by our studies with GR heterozygous mice. We propose that by inducing GCR, TNFalpha inhibits a major brake on inflammation and thereby amplifies the pro-inflammatory response. Our findings might prove helpful in understanding GCR in inflammatory diseases in which TNFalpha is intimately involved. PMID- 21646350 TI - p62 and NDP52 proteins target intracytosolic Shigella and Listeria to different autophagy pathways. AB - Autophagy is an important mechanism of innate immune defense. We have recently shown that autophagy components are recruited with septins, a new and increasingly characterized cytoskeleton component, to intracytosolic Shigella that have started to polymerize actin. On the other hand, intracytosolic Listeria avoids autophagy recognition by expressing ActA, a bacterial effector required for actin polymerization. Here, we exploit Shigella and Listeria as intracytosolic tools to characterize different pathways of selective autophagy. We show that the ubiquitin-binding adaptor proteins p62 and NDP52 target Shigella to an autophagy pathway dependent upon septin and actin. In contrast, p62 or NDP52 targets the Listeria ActA mutant to an autophagy pathway independent of septin or actin. TNF-alpha, a host cytokine produced upon bacterial infection, stimulates p62-mediated autophagic activity and restricts the survival of Shigella and the Listeria ActA mutant. These data provide a new molecular framework to understand the emerging complexity of autophagy and its ability to achieve specific clearance of intracytosolic bacteria. PMID- 21646351 TI - MicroRNA-372 is down-regulated and targets cyclin-dependent kinase 2 (CDK2) and cyclin A1 in human cervical cancer, which may contribute to tumorigenesis. AB - MicroRNAs are a class of noncoding RNAs that are ~22 nucleotides in length. MicroRNAs have been shown to play important roles in cell differentiation and in cancer. Recently, studies have shown that miR-372 is tumorigenic in human reproductive system cancers. However, we provide evidence that miR-372 acts as a tumor suppressor gene in cervical carcinoma. miR-372 was found down-regulated in cervical carcinoma tissues as compared with adjacent normal cervical tissues. Growth curve and FACS assays indicated that ectopic expression of miR-372 suppressed cell growth and induced arrest in the S/G2 phases of cell cycle in HeLa cells. We used bioinformatic predictions to determine that CDK2 and cyclin A1 were possible targets of miR-372 and confirmed this prediction using a fluorescent reporter assay. Taken together, these findings indicate that an anti oncogenic role of miR-372 may be through control of cell growth and cell cycle progression by down-regulating the cell cycle genes CDK2 and cyclin A1. PMID- 21646352 TI - Apolipoprotein L6, induced in atherosclerotic lesions, promotes apoptosis and blocks Beclin 1-dependent autophagy in atherosclerotic cells. AB - Inflammatory cytokine-regulated apoptosis and autophagy play pivotal roles in plaque rupture and thrombosis of atherosclerotic lesions. However, the molecular interplay between apoptosis and autophagy in vascular cells has not been investigated. Our prior study showed that human apolipoprotein L6 (ApoL6), a pro apoptotic BH3-only member of the Bcl-2 family, was one of the downstream targets of interferon-gamma (INFgamma), which sensitizes atherosclerotic lesion-derived cells (LDCs) to Fas-induced apoptosis. To investigate whether ApoL6 plays a causal role in atherosclerotic apoptosis and autophagy, in this study, we demonstrate that IFNgamma treatment itself strongly induces ApoL6, and ApoL6 is highly expressed and partially co-localized with activated caspase 3 in activated smooth muscle cells in atherosclerotic lesions. In addition, overexpression of ApoL6 promotes reactive oxygen species (ROS) generation, caspase activation, and subsequent apoptosis, which can be blocked by pan caspase inhibitor and ROS scavenger. Knockdown of ApoL6 expression by siApoL6 suppresses INFgamma- and Fas mediated apoptosis. Further, ApoL6 binds Bcl-X(L), one of the most abundant anti death proteins in LDCs. Interestingly, forced ApoL6 expression in LDCs induces degradation of Beclin 1, accumulation of p62, and subsequent attenuation of LC3 II formation and translocation and thus autophagy, whereas siApoL6 treatment reverts the phenotype. Taken together, our results suggest that ApoL6 regulates both apoptosis and autophagy in SMCs. IFNgamma-initiated, ApoL6-induced apoptosis in vascular cells may be an important factor causing plaque instability and a potential therapeutic target for treating atherosclerosis and cardiovascular disease. PMID- 21646353 TI - The lumenal loop Met672-Pro707 of copper-transporting ATPase ATP7A binds metals and facilitates copper release from the intramembrane sites. AB - The copper-transporting ATPase ATP7A has an essential role in human physiology. ATP7A transfers the copper cofactor to metalloenzymes within the secretory pathway; inactivation of ATP7A results in an untreatable neurodegenerative disorder, Menkes disease. Presently, the mechanism of ATP7A-mediated copper release into the secretory pathway is not understood. We demonstrate that the characteristic His/Met-rich segment Met(672)-Pro(707) (HM-loop) that connects the first two transmembrane segments of ATP7A is important for copper release. Mutations within this loop do not prevent the ability of ATP7A to form a phosphorylated intermediate during ATP hydrolysis but inhibit subsequent dephosphorylation, a step associated with copper release. The HM-loop inserted into a scaffold protein forms two structurally distinct binding sites and coordinates copper in a mixed His-Met environment with an ~2:1 stoichiometry. Binding of either copper or silver, a Cu(I) analog, induces structural changes in the loop. Mutations of 4 Met residues to Ile or two His-His pairs to Ala-Gly decrease affinity for copper. Altogether, the data suggest a two-step process, where copper released from the transport sites binds to the first His(Met)(2) site, triggering a structural change and binding to a second 2-coordinate His-His or His-Met site. We also show that copper binding within the HM-loop stabilizes Cu(I) and protects it from oxidation, which may further aid the transfer of copper from ATP7A to acceptor proteins. The mechanism of copper entry into the secretory pathway is discussed. PMID- 21646354 TI - Transportin 2 regulates apoptosis through the RNA-binding protein HuR. AB - In response to severe stress, apoptotic cell death is engaged. Apoptosis is a well orchestrated process that involves the activation and implication of many factors. In this study, we identified a role for the nuclear trafficking factor TRN2 (transportin 2) in cell death. TRN2 is normally responsible for the nuclear import of the RNA-binding protein HuR. During apoptosis, however, HuR accumulates in the cytoplasm. This is due to the caspase-mediated cleavage of the cytoplasmic fraction of HuR. One of the cleavage fragments generated by this processing of HuR interacts with TRN2 and thus blocks the re-import of HuR into the nucleus. This concentrates HuR in the cytoplasm, advancing apoptosis. Therefore, increasing or decreasing the levels of TRN2 has an inverse consequential effect on cell death, demonstrating for the first time the role of a nucleocytoplasmic transport factor in apoptosis. PMID- 21646355 TI - Cell type-specific target selection by combinatorial binding of Smad2/3 proteins and hepatocyte nuclear factor 4alpha in HepG2 cells. AB - Specific regulation of target genes by transforming growth factor-beta (TGF-beta) in a given cellular context is determined in part by transcription factors and cofactors that interact with the Smad complex. In this study, we determined Smad2 and Smad3 (Smad2/3) binding regions in the promoters of known genes in HepG2 hepatoblastoma cells, and we compared them with those in HaCaT epidermal keratinocytes to elucidate the mechanisms of cell type- and context-dependent regulation of transcription induced by TGF-beta. Our results show that 81% of the Smad2/3 binding regions in HepG2 cells were not shared with those found in HaCaT cells. Hepatocyte nuclear factor 4alpha (HNF4alpha) is expressed in HepG2 cells but not in HaCaT cells, and the HNF4alpha-binding motif was identified as an enriched motif in the HepG2-specific Smad2/3 binding regions. Chromatin immunoprecipitation sequencing analysis of HNF4alpha binding regions under TGF beta stimulation revealed that 32.5% of the Smad2/3 binding regions overlapped HNF4alpha bindings. MIXL1 was identified as a new combinatorial target of HNF4alpha and Smad2/3, and both the HNF4alpha protein and its binding motif were required for the induction of MIXL1 by TGF-beta in HepG2 cells. These findings generalize the importance of binding of HNF4alpha on Smad2/3 binding genomic regions for HepG2-specific regulation of transcription by TGF-beta and suggest that certain transcription factors expressed in a cell type-specific manner play important roles in the transcription regulated by the TGF-beta-Smad signaling pathway. PMID- 21646356 TI - Metalloprotease meprin beta generates nontoxic N-terminal amyloid precursor protein fragments in vivo. AB - Identification of physiologically relevant substrates is still the most challenging part in protease research for understanding the biological activity of these enzymes. The zinc-dependent metalloprotease meprin beta is known to be expressed in many tissues with functions in health and disease. Here, we demonstrate unique interactions between meprin beta and the amyloid precursor protein (APP). Although APP is intensively studied as a ubiquitously expressed cell surface protein, which is involved in Alzheimer disease, its precise physiological role and relevance remain elusive. Based on a novel proteomics technique termed terminal amine isotopic labeling of substrates (TAILS), APP was identified as a substrate for meprin beta. Processing of APP by meprin beta was subsequently validated using in vitro and in vivo approaches. N-terminal APP fragments of about 11 and 20 kDa were found in human and mouse brain lysates but not in meprin beta(-/-) mouse brain lysates. Although these APP fragments were in the range of those responsible for caspase-induced neurodegeneration, we did not detect cytotoxicity to primary neurons treated by these fragments. Our data demonstrate that meprin beta is a physiologically relevant enzyme in APP processing. PMID- 21646357 TI - Transmembrane helix 1 contributes to substrate translocation and protein stability of bile acid transporter SLC10A2. AB - The human apical sodium-dependent bile acid transporter (hASBT, SLC10A2) plays a critical role in the enterohepatic circulation of bile acids, as well as in cholesterol homeostasis. ASBT reclaims bile acids from the distal ileum via active sodium co-transport, in a multistep process, orchestrated by key residues in exofacial loop regions, as well as in membrane-spanning helices. Here, we unravel the functional contribution of highly conserved transmembrane helix 1 (TM1) on the hASBT transport cycle. Consecutive cysteine substitution of individual residues along the TM1 helix (Ile(29)-Gly(50)), as well as exofacial Asn(27) and Asn(28), resulted in functional impairment of ~70% of mutants, despite appreciable cell surface expression for all but G50C. Cell surface expression of G50C and G50A was rescued upon MG132 treatment as well as cyclosporine A, but not by FK506 or bile acids, suggesting that Gly(50) is involved in hASBT folding. TM1 accessibility to membrane-impermeant MTSET remains confined to the exofacial half of the helix along a single, discrete face. Substrate protection from MTSET labeling was temperature-dependent for L34C, T36C, and L38C, consistent with conformational changes playing a role in solvent accessibility for these mutants. Residue Leu(30) was shown to be critical for both bile acid and sodium affinity, while Asn(27), Leu(38), Thr(39), and Met(46) participate in sodium co-transport. Combined, our data demonstrate that TM1 plays a pivotal role in ASBT function and stability, thereby providing further insight in its dynamic transport mechanism. PMID- 21646358 TI - Distal end of carboxyl terminus is not essential for the assembly of rat Eag1 potassium channels. AB - The assembly of four pore-forming alpha-subunits into tetramers is a prerequisite for the formation of functional K(+) channels. A short carboxyl assembly domain (CAD) in the distal end of the cytoplasmic carboxyl terminus has been implicated in the assembly of Eag alpha-subunits, a subfamily of the ether-a-go-go K(+) channel family. The precise role of CAD in the formation of Eag tetrameric channels, however, remains unclear. Moreover, it has not been determined whether other protein regions also contribute to the assembly of Eag subunits. We addressed these questions by studying the biophysical properties of a series of different rat Eag1 (rEag1) truncation mutants. Two truncation mutants without CAD (K848X and W823X) yielded functional phenotypes similar to those for wild-type (WT) rEag1 channels. Furthermore, nonfunctional rEag1 truncation mutants lacking the distal region of the carboxyl terminus displayed substantial dominant negative effects on the functional expression of WT as well as K848X and W823X channels. Our co-immunoprecipitation studies further revealed that truncation mutants containing no CAD indeed displayed significant association with rEag1-WT subunits. Finally, surface biotinylation and protein glycosylation analyses demonstrated that progressive truncations of the carboxyl terminus resulted in aggravating disruptions of membrane trafficking and glycosylation of rEag1 proteins. Overall, our data suggest that the distal carboxyl terminus, including CAD, is dispensable for the assembly of rEag1 K(+) channels but may instead be essential for ensuring proper protein biosynthesis. We propose that the S6 segment and the proximal carboxyl terminus may constitute the principal subunit recognition site for the assembly of rEag1 channels. PMID- 21646359 TI - The styryl dye FM1-43 suppresses odorant responses in a subset of olfactory neurons by blocking cyclic nucleotide-gated (CNG) channels. AB - Many olfactory receptor neurons use a cAMP-dependent transduction mechanism to transduce odorants into depolarizations. This signaling cascade is characterized by a sequence of two currents: a cation current through cyclic nucleotide-gated channels followed by a chloride current through calcium-activated chloride channels. To date, it is not possible to interfere with these generator channels under physiological conditions with potent and specific blockers. In this study we identified the styryl dye FM1-43 as a potent blocker of native olfactory cyclic nucleotide-gated channels. Furthermore, we characterized this substance to stain olfactory receptor neurons that are endowed with cAMP-dependent transduction. This allows optical differentiation and pharmacological interference with olfactory receptor neurons at the level of the signal transduction. PMID- 21646360 TI - Photoprotection in plants involves a change in lutein 1 binding domain in the major light-harvesting complex of photosystem II. AB - Nonphotochemical quenching (NPQ) is the fundamental process by which plants exposed to high light intensities dissipate the potentially harmful excess energy as heat. Recently, it has been shown that efficient energy dissipation can be induced in the major light-harvesting complexes of photosystem II (LHCII) in the absence of protein-protein interactions. Spectroscopic measurements on these samples (LHCII gels) in the quenched state revealed specific alterations in the absorption and circular dichroism bands assigned to neoxanthin and lutein 1 molecules. In this work, we investigate the changes in conformation of the pigments involved in NPQ using resonance Raman spectroscopy. By selective excitation we show that, as well as the twisting of neoxanthin that has been reported previously, the lutein 1 pigment also undergoes a significant change in conformation when LHCII switches to the energy dissipative state. Selective two photon excitation of carotenoid (Car) dark states (Car S(1)) performed on LHCII gels shows that the extent of electronic interactions between Car S(1) and chlorophyll states correlates linearly with chlorophyll fluorescence quenching, as observed previously for isolated LHCII (aggregated versus trimeric) and whole plants (with versus without NPQ). PMID- 21646362 TI - Material vs. psychosocial explanations of old-age educational differences in physical and mental functioning. AB - BACKGROUND: Taking into account our rapidly ageing population, older people are of particular interest in studying health inequalities. The aim of the present study is to examine the relation between socio-economic status and health-related functioning in older people and to find out how material factors (e.g. the lack of basic goods) and psychosocial factors (e.g. low self-efficacy) compare regarding the explanation of these socio-economic differences. METHODS: Data came from 5061 Dutch men and women aged >= 55 years who participated in the longitudinal Study on Medical Information and Lifestyles Eindhoven (SMILE) study. Baseline data were collected between November 2002 and May 2004 and respondents were followed until May 2009 (follow-up range: 0-5 years). Multilevel analyses were used to study the association between educational level and longitudinal changes in physical and mental functioning (i.e. two subscales of the SF-36) and to study the relative contribution of material and psychosocial factors to this relation. RESULTS: Low educational level was associated with poor initial physical and mental functioning. However, no further widening of these gradients was found during follow-up. Material factors reduced the initial educational differences by an average of 29%, whereas psychosocial factors, mastery and self efficacy in particular, reduced these differences by an average of 60%. CONCLUSION: More than material factors, psychosocial factors, mastery and self efficacy in particular, explained a large part of the educational differences in physical and mental functioning in older people. Further research is recommended to explore the amenability to change of characteristics that hamper people from taking control over their lives. PMID- 21646361 TI - Ultraviolet irradiation can induce evasion of colon cancer cells from stimulation of epidermal growth factor. AB - Receptor down-regulation is the most prominent regulatory system of EGF receptor (EGFR) signal attenuation and a critical target for therapy against colon cancer, which is highly dependent on the function of the EGFR. In this study, we investigated the effect of ultraviolet-C (UV-C) on down-regulation of EGFR in human colon cancer cells (SW480, HT29, and DLD-1). UV-C caused inhibition of cell survival and proliferation, concurrently inducing the decrease in cell surface EGFR and subsequently its degradation. UV-C, as well as EGFR kinase inhibitors, decreased the expression level of cyclin D1 and the phosphorylated level of retinoblastoma, indicating that EGFR down-regulation is correlated to cell cycle arrest. Although UV-C caused a marked phosphorylation of EGFR at Ser-1046/1047, UV-C also induced activation of p38 MAPK, a stress-inducible kinase believed to negatively regulate tumorigenesis, and the inhibition of p38 MAPK canceled EGFR phosphorylation at Ser-1046/1047, as well as subsequent internalization and degradation, suggesting that p38 MAPK mediates EGFR down-regulation by UV-C. In addition, phosphorylation of p38 MAPK induced by UV-C was mediated through transforming growth factor-beta-activated kinase-1. Moreover, pretreatment of the cells with UV-C suppressed EGF-induced phosphorylation of EGFR at tyrosine residues in addition to cell survival signal, Akt. Together, these results suggest that UV-C irradiation induces the removal of EGFRs from the cell surface that can protect colon cancer cells from oncogenic stimulation of EGF, resulting in cell cycle arrest. Hence, UV-C might be applied for clinical strategy against human colon cancers. PMID- 21646363 TI - Social inequalities in obesity and overweight in 11 OECD countries. AB - BACKGROUND: Evidence of inequalities in obesity and overweight is available mostly from national studies. This article provides a broad international comparison of inequalities by education level and socio-economic status, in men and women and over time. METHODS: Data from national health surveys of 11 OECD countries were used. The size of inequalities was assessed on the basis of absolute and relative inequality indexes. A regression-analysis approach was used to assess differences between social groups in trends over time. RESULTS: Of the countries examined, USA and England had the highest rates of obesity and overweight. Large social inequalities were consistently detected in all countries, especially in women. Absolute inequalities were largest in Hungary and Spain with a difference of 11.6 and 10% in obesity rates in men, and 18.3 and 18.9% in women, respectively, across the education spectrum. Relative inequalities were largest in France and Sweden with poorly educated men 3.2 and 2.8 times as likely to be obese as men with the highest education (18 and 17 times for women in Spain and Korea, respectively). Pro-poor inequalities in overweight were observed for men in USA, Canada, Korea, Hungary, Australia and England. Inequalities remained virtually stable during the last 15 years, with only small variations in England, Korea, Italy and France. CONCLUSION: Large and persistent social inequalities in obesity and overweight by education level and socio-economic status exist in OECD countries. These are consistently larger in women than in men. PMID- 21646364 TI - Subjective social status: its determinants and association with health in the Swedish working population (the SLOSH study). AB - BACKGROUND: This study aims to confirm associations between subjective social status (SSS) and health in the Swedish working population, to investigate if SSS is related to health over and above conventional measures of socio-economic status and to identify factors related to perception of SSS. METHODS: The study includes 2358 men and 2665 women in the age group of 19-68 years who participated in the Swedish Longitudinal Occupational Survey of Health (SLOSH). SSS was measured using the MacArthur scale, a 10-rung ladder on which individuals indicate where they think they stand in the social hierarchy. Health, health behaviors and social situation were also measured in 2006, while more objective measures of socio-economic position were derived from registry data and interviews in 2003. RESULTS: Perception of SSS was cross-sectionally related to age-adjusted prevalence of suboptimal perceived general health, sleep disturbances, musculoskeletal symptoms, depressive symptoms and emotional exhaustion. Relative Indices of Inequalities (RII) showed significant relationships between SSS and health, which were attenuated by adjustment for general life satisfaction. Adjustment for occupational grade, education and income further attenuated the relationship. The main predictors of SSS in 2006 were occupational grade, personal income and education in 2003; household financial situation, general life satisfaction and job control in 2006. When ranking their SSS, women put more weight on household financial situation and men on their personal income. CONCLUSION: SSS is associated with prevalence of several health complaints in the Swedish working population over and above conventional SES, indicating that SSS is a valid measure of social inequality in health. PMID- 21646365 TI - Evaluation of the impact of a smoke-free home initiative in Rotherham, a deprived district in Northern England. AB - BACKGROUND: An evaluation of a smoke-free home initiative launched in Rotherham, northern England, in July 2009. METHODS: Two approaches were used: (i) a postal survey of participants 4 months after signing up as a SFH and (ii) a telephone consultation. The survey was sent to 620 households (of 654 who signed up to the scheme); 289 (46.6%) were returned. The telephone consultation involved 20 households before and 20 after signing up to the scheme. RESULTS: Of the households that permitted some smoking at home before the initiative, ~78% became smoke free after signing up (uncertainty due to missing replies). A high number of participants (169, 60.8%) were already informally smoke free. The most common reasons for participation concerned health, environment, and fire safety. Participants were motivated by, amongst other things, information given in a booklet and by the offer of a fire-safety referral. The most immediate benefits noted by participants were improvements in house hygiene. The most important hindrance to success seemed to be a lack of power to enforce the ban at home, particularly on the part of those living in smokers' homes. CONCLUSION: The Rotherham initiative succeeded in creating smoke-free homes. The results should help those planning similar initiatives. Important points include that: many participants had already instituted some rules regarding smoking at home; whether and how to include households that are already smoke-free; risk of fire and concern with house hygiene are important motivations; those living in smokers' homes may lack power to initiate smoke-free rules. PMID- 21646366 TI - A-Z of nutritional supplements: dietary supplements, sports nutrition foods and ergogenic aids for health and performance--Part 22. PMID- 21646367 TI - Evidences for the existence of a low dopaminergic tone in polycystic ovarian syndrome: implications for OHSS development and treatment. AB - CONTEXT: The dopamine/dopamine receptor 2 (D2/Drd2) pathway modulates vascular endothelial growth factor (VEGF)-dependent vascular permeability and angiogenesis in the ovary. Deregulation of the VEGF/VEGF receptor (VEGFR)-2 pathway leading to increased risk of ovarian hyperstimulation syndrome has been described in the ovary of patients suffering from polycystic ovarian syndrome (PCOS). OBJECTIVE: The objective of the study was to ascertain whether deregulation of the VEGF/VEGFR-2 might a least be partially due to abnormalities of the D2/Drd2 pathway in PCOS women. DESIGN: Dated, archived ovaries from PCOs and control group patients as well as human chorionic gonadotropin-stimulated luteinized granulosa cells form PCOS and non-PCOS oocyte patients were used. SETTING: The study was conducted at a private research center. PATIENTS OR OTHER PARTICIPANTS: PCOS and nonpolycystic ovarian patients and oocyte patients participated in the study. INTERVENTION(S): Human ovarian sections were stained against the Drd2 antibody. Human chorionic gonadotropin-stimulated luteinized granulosa cells (LGC) were cultured in the presence/absence and the Drd2 agonist cabergoline. MAIN OUTCOME MEASURE(S): Drd2 and vascularized stained area in the theca layer of antral (< 8 mm) and luteinized follicles was quantified. VEGF, D2, and its related metabolites were measured in the supernatant of cultured LGC by ELISA and HPLC, respectively. VEGFR-2 and Drd2 expressed by LGC was quantified through an In-Cell ELISA. RESULTS: Decreased Drd2 expression and increased vascularization in the theca layer of antral and luteinized follicles of PCOS ovaries was observed. A lower dopamine production and reduced efficacy of cabergoline in inhibiting VEGF secretion was uncovered in LGC from PCOS. CONCLUSIONS: Decreased dopaminergic tone as well as deregulated Drd2 signaling might explain higher VEGF and vascularization leading to increased ovarian hyperstimulation syndrome risk in PCOS. PMID- 21646368 TI - Evaluation, treatment, and prevention of vitamin D deficiency: an Endocrine Society clinical practice guideline. AB - OBJECTIVE: The objective was to provide guidelines to clinicians for the evaluation, treatment, and prevention of vitamin D deficiency with an emphasis on the care of patients who are at risk for deficiency. PARTICIPANTS: The Task Force was composed of a Chair, six additional experts, and a methodologist. The Task Force received no corporate funding or remuneration. CONSENSUS PROCESS: Consensus was guided by systematic reviews of evidence and discussions during several conference calls and e-mail communications. The draft prepared by the Task Force was reviewed successively by The Endocrine Society's Clinical Guidelines Subcommittee, Clinical Affairs Core Committee, and cosponsoring associations, and it was posted on The Endocrine Society web site for member review. At each stage of review, the Task Force received written comments and incorporated needed changes. CONCLUSIONS: Considering that vitamin D deficiency is very common in all age groups and that few foods contain vitamin D, the Task Force recommended supplementation at suggested daily intake and tolerable upper limit levels, depending on age and clinical circumstances. The Task Force also suggested the measurement of serum 25-hydroxyvitamin D level by a reliable assay as the initial diagnostic test in patients at risk for deficiency. Treatment with either vitamin D(2) or vitamin D(3) was recommended for deficient patients. At the present time, there is not sufficient evidence to recommend screening individuals who are not at risk for deficiency or to prescribe vitamin D to attain the noncalcemic benefit for cardiovascular protection. PMID- 21646369 TI - Persistence of sleep-associated decrease in GnRH pulse frequency in the absence of gonadal steroids. AB - CONTEXT: There is dramatic slowing of GnRH pulse frequency during sleep in the early follicular phase of the menstrual cycle, but it is unknown whether this represents a primary effect of sleep or is dependent upon the sex steroid environment. OBJECTIVES: Our objective was to determine 1) whether sleep affects GnRH pulse frequency in postmenopausal women (PMW) in whom gonadal hormones are low and 2) whether this relationship changes with aging. DESIGN AND SETTING: Studies were performed in the Clinical Research Center of an academic medical center. SUBJECTS: Subjects included healthy PMW, 45-55 (n = 8) and 70-80 (n = 6) years old. INTERVENTIONS: Subjects were studied during one night of polysomnographic-recorded sleep and one night of monitored wake during which blood was sampled every 5 min for 8 h. MAIN OUTCOME MEASURES: Pulsatile secretion of free alpha-subunit (FAS), a marker of GnRH secretion, was assessed. RESULTS: There were no differences in sleep macroarchitecture or sleep efficiency [75 +/- 12% (mean +/- sd)] between older and younger PMW. The FAS interpulse interval was longer during sleep than nighttime wake in all women (60.5 +/- 4.3 vs. 52.0 +/- 2.8 min, P = 0.03) with a similar effect in the two groups. FAS pulse amplitude did not differ between sleep and wake periods (474.8 +/- 36.7 vs. 478.2 +/- 36.5 ng/liter, P = 0.9). CONCLUSIONS: Sleep is associated with a significant decline in GnRH pulse frequency in both older and younger PMW. Its persistence in PMW reinforces the important connection between sleep and GnRH secretion. PMID- 21646370 TI - Role of glycated hemoglobin in the prediction of future risk of T2DM. AB - AIM: The aim of this study was to assess the predictive power of glycated hemoglobin (HbA1c) for future type 2 diabetes risk. RESEARCH DESIGN AND METHODS: Six hundred eighty-seven subjects who were free of type 2 diabetes mellitus (T2DM) participated in the study. Each subject received a 75-g oral glucose tolerance test at baseline and 624 received a repeat oral glucose tolerance test after 3.5 +/- 0.1 yr of follow-up. Anthropometric measurements, lipid profile, and HbA1c were measured during the baseline visit. Logistic multivariate models were created with T2DM status at follow-up as the dependent variable and other parameters as the independent variables. The receiver-operating characteristic (ROC) was used to assess the predictive discrimination of the various models. RESULTS: HbA1c was a significant predictor of future T2DM risk (area under the ROC curve = 0.73, P < 0.0001). A HbA1c cut point of 5.65% had the maximal sum of sensitivity and specificity. Although the area under the ROC curve of HbA1c was smaller than the area under the ROC curve of both the 1-h plasma glucose concentration and a multivariate logistic model (including anthropometric parameters, lipid profile, and fasting plasma glucose), the addition of HbA1c to both the 1-h plasma glucose and the multivariate logistic model significantly increased their predictive power. CONCLUSION: Although HbA1c alone is a weaker predictor of future T2DM risk compared with the 1-h plasma glucose, it provides additive information about future T2DM risk when added to previously published prediction models. PMID- 21646371 TI - Nephrolithiasis and renal calcifications in primary hyperparathyroidism. AB - CONTEXT: Renal complications in terms of hypercalciuria, nephrolithiasis, and nephrocalcinosis are well-known risks in primary hyperparathyroidism (PHPT) and may lead to impaired renal function. EVIDENCE ACQUISITION: We reviewed published evidence on the occurrence, pathophysiology, and consequences of renal complications in PHPT and highlighted areas of uncertainty that should be investigated further. EVIDENCE SYNTHESIS: In asymptomatic PHPT, renal stones are present in approximately 7% of the patients, which is a significantly higher prevalence than among patients without PHPT (1.6%). Also, before diagnosis of PHPT, risk of hospital admissions due to renal stones is increased compared with the background population, and the risk remains increased for at least 10 yr after surgical cure from PHPT. However, shortly after parathyroidectomy, risk of recurrent stone episodes is reduced to the recurrence rate among patients with idiopathic renal stone disease. In general, patients with PHPT who develop nephrolithiasis are of younger age and more often are males, compared with those who do not form renal calcifications. Although 24-h urinary calcium is decreased after parathyroidectomy, studies have shown a higher renal calcium excretion and lower serum phosphate levels in former PHPT patients compared with healthy controls, suggesting that these patients have some additional mineral disorder. CONCLUSION: All patients with a diagnosis of PHPT should initially be evaluated for renal calcifications by unenhanced helical computed tomography. If calcifications are present, parathyroidectomy is recommended. If symptoms develop after parathyroidectomy, patients should be evaluated and treated similar to other patients with renal stones. PMID- 21646372 TI - Low testosterone in men with type 2 diabetes: significance and treatment. AB - CONTEXT: The relationship between testosterone and diabetes in men is an important issue, given that one third of U.S. men aged 65 yr or older have diabetes, with a similar percentage having low testosterone levels. EVIDENCE ACQUISITION: The medical literature from 1970 to March 2011 was reviewed for key articles. EVIDENCE SYNTHESIS: In population-based studies, low testosterone is commonly associated with type 2 diabetes and the metabolic syndrome, and it identifies men with an adverse metabolic profile. The difference in testosterone levels between men with diabetes compared to men without diabetes is moderate and comparable in magnitude to the effects of other chronic diseases, suggesting that low testosterone may be a marker of poor health. Although the inverse association of testosterone with diabetes is partially mediated by SHBG, low testosterone is linked to diabetes via a bidirectional relationship with visceral fat, muscle, and possibly bone. There is consistent evidence from randomized trials that testosterone therapy alters body composition in a metabolically favorable manner, but changes are modest and have not consistently translated into reductions in insulin resistance or improvements in glucose metabolism. CONCLUSIONS: The key response to the aging, overweight man with type 2 diabetes and subnormal testosterone levels should be implementation of lifestyle measures such as weight loss and exercise, which, if successful, raise testosterone and provide multiple health benefits. Although approved therapy for diabetes should be used, testosterone therapy should not be given to such men until benefits and risks are clarified by adequately powered clinical trials. PMID- 21646373 TI - Senescent cells develop a PARP-1 and nuclear factor-{kappa}B-associated secretome (PNAS). AB - Melanoma cells can enter the process of senescence, but whether they express a secretory phenotype, as reported for other cells, is undetermined. This is of paramount importance, because this secretome can alter the tumor microenvironment and the response to chemotherapeutic drugs. More generally, the molecular events involved in formation of the senescent-associated secretome have yet to be determined. We reveal here that melanoma cells experiencing senescence in response to diverse stimuli, including anti-melanoma drugs, produce an inflammatory secretory profile, where the chemokine ligand-2 (CCL2) acts as a critical effector. Thus, we reveal how senescence induction might be involved in therapeutic failure in melanoma. We further provide a molecular relationship between senescence induction and secretome formation by revealing that the poly(ADP-ribose) polymerase-1 (PARP-1)/nuclear factor-kappaB (NF-kappaB) signaling cascade, activated during senescence, drives the formation of a secretome endowed with protumoral and prometastatic properties. Our findings also point to the existence of the PARP-1 and NF-kappaB-associated secretome, termed the PNAS, in nonmelanoma cells. Most importantly, inhibition of PARP-1 or NF kappaB prevents the proinvasive properties of the secretome. Collectively, identification of the PARP-1/NF-kappaB axis in secretome formation opens new avenues for therapeutic intervention against cancers. PMID- 21646374 TI - Separation of the gluconeogenic and mitochondrial functions of PGC-1{alpha} through S6 kinase. AB - PGC-1alpha is a transcriptional coactivator that powerfully regulates many pathways linked to energy homeostasis. Specifically, PGC-1alpha controls mitochondrial biogenesis in most tissues but also initiates important tissue specific functions, including fiber type switching in skeletal muscle and gluconeogenesis and fatty acid oxidation in the liver. We show here that S6 kinase, activated in the liver upon feeding, can phosphorylate PGC-1alpha directly on two sites within its arginine/serine-rich (RS) domain. This phosphorylation significantly attenuates the ability of PGC-1alpha to turn on genes of gluconeogenesis in cultured hepatocytes and in vivo, while leaving the functions of PGC-1alpha as an activator of mitochondrial and fatty acid oxidation genes completely intact. These phosphorylations interfere with the ability of PGC 1alpha to bind to HNF4alpha, a transcription factor required for gluconeogenesis, while leaving undisturbed the interactions of PGC-1alpha with ERRalpha and PPARalpha, factors important for mitochondrial biogenesis and fatty acid oxidation. These data illustrate that S6 kinase can modify PGC-1alpha and thus allow molecular dissection of its functions, providing metabolic flexibility needed for dietary adaptation. PMID- 21646375 TI - Anatomic localization and autonomic modulation of atrioventricular junctional rhythm in failing human hearts. AB - BACKGROUND: The structure-function relationship in the atrioventricular junction (AVJ) of various animal species has been investigated in detail; however, less is known about the human AVJ. In this study, we performed high-resolution optical mapping of the human AVJ (n = 6) to define its pacemaker properties and response to autonomic stimulation. METHODS AND RESULTS: Isolated, coronary-perfused AVJ preparations from failing human hearts (n = 6, 53 +/- 6 years) were optically mapped using the near-infrared, voltage-sensitive dye, di-4-ANBDQBS, with isoproterenol (1 MUmol/L) and acetylcholine (1 MUmol/L). An algorithm detecting multiple components of optical action potentials was used to reconstruct multilayered intramural AVJ activation and to identify specialized slow and fast conduction pathways (SP and FP). The anatomic origin and propagation of pacemaker activity was verified by histology. Spontaneous AVJ rhythms of 29 +/- 11 bpm (n = 6) originated in the nodal-His region (n = 3) and/or the proximal His bundle (n = 4). Isoproterenol accelerated the AVJ rhythm to 69 +/- 12 bpm (n = 5); shifted the leading pacemaker to the transitional cell regions near the FP and SP (n = 4) and/or coronary sinus (n = 2); and triggered reentrant arrhythmias (n = 2). Acetylcholine (n = 4) decreased the AVJ rhythm to 18 +/- 4 bpm; slowed FP/SP conduction leading to block between the AVJ and atrium; and shifted the pacemaker to either the transitional cell region or the nodal-His region (bifocal activation). CONCLUSIONS: We have demonstrated that the AVJ pacemaker in failing human hearts is located in the nodal-His region or His bundle regions and can be modified with autonomic stimulation. Moreover, we found that both the FP and SP are involved in anterograde and retrograde conduction. PMID- 21646376 TI - Choosing a mouse model: experimental biology in context--the utility and limitations of mouse models of breast cancer. AB - Genetically engineered mice are critical experimental models for the study of breast cancer biology. Transgenic mice, employing strong mammary epithelial promoters to drive oncogenes, develop carcinomas with phenotypes corresponding to the molecular pathway activated. Gene-targeted (knockout) mice, in which tumor suppressors are deleted, develop mammary neoplasms with phenotypes primarily including patterns seen in spontaneous mouse mammary tumors, albeit at higher rates. Improved genetic engineering, using inducible gene expression, somatic gene transduction, conditional alleles, and crossbreeding for combined/compound genetic engineering yields precise molecular models with exquisite experimental control and phenotypes with comparative pathologic validity. Mammary gland transplantation technology adds a practical and validated method for assessing biologic behavior of selected mammary tissues. Overall, the many mouse models available are a rich resource for experimental biology with phenocopies of breast cancer subtypes, and a variety of practical advantages. The challenge is matching the model to the experimental question. PMID- 21646377 TI - Molecular regulation of the mitosis/meiosis decision in multicellular organisms. AB - A major step in the journey from germline stem cell to differentiated gamete is the decision to leave the mitotic cell cycle and begin progression through the meiotic cell cycle. Over the past decade, molecular regulators of the mitosis/meiosis decision have been discovered in most of the major model multicellular organisms. Historically, the mitosis/meiosis decision has been closely linked with controls of germline self-renewal and the sperm/egg decision, especially in nematodes and mice. Molecular explanations of those linkages clarify our understanding of this fundamental germ cell decision, and unifying themes have begun to emerge. Although the complete circuitry of the decision is not known in any organism, the recent advances promise to impact key issues in human reproduction and agriculture. PMID- 21646378 TI - Lipid polymorphisms and membrane shape. AB - Morphological plasticity of biological membrane is critical for cellular life, as cells need to quickly rearrange their membranes. Yet, these rearrangements are constrained in two ways. First, membrane transformations may not lead to undesirable mixing of, or leakage from, the participating cellular compartments. Second, membrane systems should be metastable at large length scales, ensuring the correct function of the particular organelle and its turnover during cellular division. Lipids, through their ability to exist with many shapes (polymorphism), provide an adequate construction material for cellular membranes. They can self assemble into shells that are very flexible, albeit hardly stretchable, which allows for their far-reaching morphological and topological behaviors. In this article, we will discuss the importance of lipid polymorphisms in the shaping of membranes and its role in controlling cellular membrane morphology. PMID- 21646380 TI - Impact of the genetic screening revolution: understanding and meeting the needs of previvors with a known family mutation in BRCA/BRCA2. PMID- 21646379 TI - Evolution and diversity of the Golgi. AB - The Golgi is an ancient and fundamental eukaryotic organelle. Evolutionary cell biological studies have begun establishing the repertoire, processes, and level of complexity of membrane-trafficking machinery present in early eukaryotic cells. This article serves as a review of the literature on the topic of Golgi evolution and diversity and reports a novel comparative genomic survey addressing Golgi machinery in the widest taxonomic diversity of eukaryotes sampled to date. Finally, the article is meant to serve as a primer on the rationale and design of evolutionary cell biological studies, hopefully encouraging readers to consider this approach as an addition to their cell biological toolbox. It is clear that the major machinery involved in vesicle trafficking to and from the Golgi was already in place by the time of the divergence of the major eukaryotic lineages, nearly 2 billion years ago. Much of this complexity was likely generated by an evolutionary process involving gene duplication and coevolution of specificity encoding membrane-trafficking proteins. There have also been clear cases of loss of Golgi machinery in some lineages as well as innovation of novel machinery. The Golgi is a wonderfully complex and diverse organelle and its continued exploration promises insight into the evolutionary history of the eukaryotic cell. PMID- 21646381 TI - A characterisation of factors determining postoperative ileus after laparoscopic colectomy enables the generation of a novel predictive score. PMID- 21646382 TI - Mixed methods evaluation of in-hospital nurse prescribing finds similar care standards and provision between nurses and doctors. PMID- 21646383 TI - Skin protection wheelchair cushions for older nursing home residents reduce 6 month incidence of ischial tuberosity pressure ulcers compared with segmented foam cushions. PMID- 21646384 TI - GnRH antagonists are safer than agonists: an update of a Cochrane review. PMID- 21646385 TI - Proteasome activator PA28{gamma} stimulates degradation of GSK3-phosphorylated insulin transcription activator MAFA. AB - MAFA is a member of the MAF family of basic leucine zipper transcription factors and is a critical regulator of insulin gene expression and islet beta-cell function. To be degraded by the proteasome, MAFA must be phosphorylated by GSK3 and MAP kinases at multiple serine and threonine residues (Ser49, Thr53, Thr57, Ser61, and Ser65) within its amino-terminal domain. In this study, we report that MAFA degradation is stimulated by PA28gamma (REGgamma and PSME3), a member of a family of proteasome activators that bind and activate the 20S proteasome. To date, only a few PA28gamma-proteasome pathway substrates have been identified, including steroid receptor coactivator 3 (SRC3) and the cell cycle inhibitor p21 (CIP1). PA28gamma binds to MAFA, induces its proteasomal degradation, and thereby attenuates MAFA-driven transcriptional activation of the insulin promoter. Co expression of GSK3 enhanced the PA28gamma-mediated degradation of MAFA, but mutants that contained alanine substitutions at the MAFA phosphorylation sites did not bind PA28gamma and were resistant to degradation. We also found that a PA28gamma mutant (N151Y) that did not stimulate p21 degradation enhanced MAFA degradation, and another mutant (K188D) that promoted greater p21 degradation did not enhance MAFA degradation. These results suggest that PA28gamma stimulates MAFA degradation through a novel molecular mechanism that is distinct from that for the degradation of p21. PMID- 21646386 TI - The effect of 17{beta}-estradiol on cholesterol content in human macrophages is influenced by the lipoprotein milieu. AB - Estrogen and testosterone are thought to modulate coronary heart disease (CHD) risk. To examine how these hormones affect human macrophage cholesterol transport, a key factor in atherogenesis, we obtained monocytes from healthy male and postmenopausal female donors (age 50-70 years). Cells were allowed to differentiate in autologous serum. Human monocyte-derived macrophages (HMDMs) were exposed to estrogen, testosterone, or vehicle, during differentiation. Cells were cholesterol enriched with oxidized low-density lipoprotein (oxLDL) in the presence of treatment. Cell cholesterol mass, efflux, and the expression of proteins involved in HMDM cholesterol transport were examined. Estrogen significantly reduced cholesteryl ester (CE) content in both female and male HMDMs while having no measurable effect on cholesterol efflux. Testosterone did not affect cholesterol content or efflux. Both hormones significantly but modestly affected the gene expression of several proteins involved in HMDM transport, yet these effects did not translate into significant changes in protein expression. In THP-1 macrophages, the effect of estrogen on CE content was more potent in unloaded macrophages and was estrogen receptor dependent. A trend for a reduction in non-oxLDL uptake by estrogen was observed and was also found to be dependent upon estrogen receptor activation. Our data indicate that estrogen, but not testosterone, reduces CE accumulation in HMDMs obtained from a CHD age relevant population, independent of changes in the expression of proteins important to macrophage cholesterol transport. In THP-1 cells, this effect is reduced in the presence of oxLDL, indicating that a pro-atherogenic lipoprotein milieu is an important variable in sex hormone modulation of CHD. PMID- 21646387 TI - Oxidative stress mediates through apoptosis the anticancer effect of phospho nonsteroidal anti-inflammatory drugs: implications for the role of oxidative stress in the action of anticancer agents. AB - We assessed the relationship between oxidative stress, cytokinetic parameters, and tumor growth in response to novel phospho-nonsteroidal anti-inflammatory drugs (NSAIDs), agents with significant anticancer effects in preclinical models. Compared with controls, in SW480 colon and MCF-7 breast cancer cells, phospho sulindac, phospho-aspirin, phospho-flurbiprofen, and phospho-ibuprofen (P-I) increased the levels of reactive oxygen and nitrogen species (RONS) and decreased GSH levels and thioredoxin reductase activity, whereas the conventional chemotherapeutic drugs (CCDs), 5-fluorouracil (5-FU), irinotecan, oxaliplatin, chlorambucil, paclitaxel, and vincristine, did not. In both cell lines, phospho NSAIDs induced apoptosis and inhibited cell proliferation much more potently than CCDs. We then treated nude mice bearing SW480 xenografts with P-I or 5-FU that had an opposite effect on RONS in vitro. Compared with controls, P-I markedly suppressed xenograft growth, induced apoptosis in the xenografts (8.9 +/- 2.7 versus 19.5 +/- 3.0), inhibited cell proliferation (52.6 +/- 5.58 versus 25.8 +/- 7.71), and increased urinary F2-isoprostane levels (10.7 +/- 3.3 versus 17.9 +/- 2.2 ng/mg creatinine, a marker of oxidative stress); all differences were statistically significant. 5-FU's effects on tumor growth, apoptosis, proliferation, and F2-isoprostane were not statistically significant. F2 isoprostane levels correlated with the induction of apoptosis and the inhibition of cell growth. P-I induced oxidative stress only in the tumors, and its apoptotic effect was restricted to xenografts. Our data show that phospho-NSAIDs act against cancer through a mechanism distinct from that of various CCDs, underscore the critical role of oxidative stress in their effect, and indicate that pathways leading to oxidative stress may be useful targets for anticancer strategies. PMID- 21646388 TI - Macrophage deletion of SOCS1 increases sensitivity to LPS and palmitic acid and results in systemic inflammation and hepatic insulin resistance. AB - OBJECTIVE: Macrophage secretion of proinflammatory cytokines contributes to the pathogenesis of obesity-related insulin resistance. An important regulator of inflammation is the suppressor of cytokine signaling-1 (SOCS1), which inhibits the JAK-STAT and toll-like receptor-4 (TLR4) pathways. Despite the reported role of SOCS1 in inhibiting insulin signaling, it is surprising that a SOCS1 polymorphism that increases SOCS1 promoter activity is associated with enhanced insulin sensitivity despite obesity. In the current study, we investigated the physiological role of myeloid and lymphoid cell SOCS1 in regulating inflammation and insulin sensitivity. RESEARCH DESIGN AND METHODS: We used mice generated by crossing SOCS1 floxed mice with mice expressing Cre recombinase under the control of the LysM-Cre promoter (SOCS1 LysM-Cre). These mice have deletion of SOCS1 in macrophages and lymphocytes. We assessed macrophage inflammation using flow cytometry and serum cytokine levels using Bioplex assays. We then measured insulin sensitivity using glucose tolerance tests and the euglycemic hyperinsulinemic clamp. Using bone marrow-derived macrophages, we tested the effects of SOCS1 deletion in regulating responses to the TLR4 ligands: lipopolysaccharide (LPS) and palmitic acid. RESULTS: SOCS1 LysM-Cre mice had increased macrophage expression of CD11c, enhanced sensitivity to LPS, and palmitic acid and increased serum concentrations of tumor necrosis factor-alpha, interleukin-6, and monocyte chemoattractant protein. Increased inflammation was associated with impaired glucose tolerance and hyperinsulinemia as a result of reduced hepatic but not skeletal muscle insulin sensitivity. CONCLUSIONS: The expression of SOCS1 in hematopoietic cells protects mice against systemic inflammation and hepatic insulin resistance potentially by inhibiting LPS and palmitate-induced TLR4 signaling in macrophages. PMID- 21646389 TI - Severity of diabetes governs vascular lipoprotein lipase by affecting enzyme dimerization and disassembly. AB - OBJECTIVE: In diabetes, when glucose consumption is restricted, the heart adapts to use fatty acid (FA) exclusively. The majority of FA provided to the heart comes from the breakdown of circulating triglyceride (TG), a process catalyzed by lipoprotein lipase (LPL) located at the vascular lumen. The objective of the current study was to determine the mechanisms behind LPL processing and breakdown after moderate and severe diabetes. RESEARCH DESIGN AND METHODS: To induce acute hyperglycemia, diazoxide, a selective, ATP-sensitive K(+) channel opener was used. For chronic diabetes, streptozotocin, a beta-cell-specific toxin was administered at doses of 55 or 100 mg/kg to generate moderate and severe diabetes, respectively. Cardiac LPL processing into active dimers and breakdown at the vascular lumen was investigated. RESULTS: After acute hyperglycemia and moderate diabetes, more LPL is processed into an active dimeric form, which involves the endoplasmic reticulum chaperone calnexin. Severe diabetes results in increased conversion of LPL into inactive monomers at the vascular lumen, a process mediated by FA-induced expression of angiopoietin-like protein 4 (Angptl 4). CONCLUSIONS: In acute hyperglycemia and moderate diabetes, exaggerated LPL processing to dimeric, catalytically active enzyme increases coronary LPL, delivering more FA to the heart when glucose utilization is compromised. In severe chronic diabetes, to avoid lipid oversupply, FA-induced expression of Angptl-4 leads to conversion of LPL to inactive monomers at the coronary lumen to impede TG hydrolysis. Results from this study advance our understanding of how diabetes changes coronary LPL, which could contribute to cardiovascular complications seen with this disease. PMID- 21646390 TI - Defective differentiation of regulatory FoxP3+ T cells by small-intestinal dendritic cells in patients with type 1 diabetes. AB - OBJECTIVE: The gut environment modulates the pathogenesis of type 1 diabetes (T1D), but how it affects autoimmunity toward pancreatic beta-cells, a self tissue located outside the intestine, is still unclear. In the small intestine, lamina propria dendritic cells (LPDCs) induce peripheral differentiation of FoxP3(+) regulatory T (Treg) cells. We tested the hypothesis that the intestinal milieu impinges on human T1D by affecting differentiation of FoxP3(+) Treg cells. RESEARCH DESIGN AND METHODS: We collected duodenal biopsies of 10 T1D patients, 16 healthy subjects, and 20 celiac individuals and performed a fluorescent activated cell sorter analysis to measure percentages of various immune cell subsets, including CD4(+) and CD8(+) T cells, NK cells, gammadelta T cells, CD103(+)CD11c(+) LPDCs, and CD4(+)CD25(+)FoxP3(+)CD127(-) Treg cells. In parallel, we assessed the tolerogenic function (i.e., capacity to induce differentiation of FoxP3(+) Treg cells) by LPDCs of T1D patients and control subjects. RESULTS: Our analysis revealed a significant reduction in the percentage of intestinal CD4(+)CD25(+)FoxP3(+)CD127(-) Treg cells in T1D patients compared with healthy subjects (P = 0.03) and celiac individuals (P = 0.003). In addition, we found that LPDCs from T1D patients completely lacked their tolerogenic function; they were unable to convert CD4(+)CD25(-) T cells into CD4(+)CD25(+)FoxP3(+)CD127(-) Treg cells. CONCLUSIONS: Our data indicate that T1D patients have a reduced number of intestinal FoxP3(+) Treg cells as a result of their defective differentiation in the gut. These findings suggest that intestinal immune regulation is not only calibrated to tolerate commensal bacteria and food components but also is instrumental in maintaining immune tolerance toward pancreatic beta-cells and preventing T1D. PMID- 21646391 TI - Excessive ovarian production of nerve growth factor elicits granulosa cell apoptosis by setting in motion a tumor necrosis factor alpha/stathmin-mediated death signaling pathway. AB - Excessive nerve growth factor (NGF) production by the ovary, achieved via a transgenic approach, results in arrested antral follicle growth, reduced ovulatory capacity, and a predisposition to cyst formation in response to mildly elevated LH levels. Two salient features in these mutant mice (termed 17NF) are an elevated production of 17alpha-hydroxyprogesterone (17-OHP(4)), testosterone, and estradiol (E(2)) in response to gonadotropins, and an increased frequency of granulosa cell (GC) apoptosis. In this study, we show that the increase in steroidal response is associated with enhanced expression of Cyp17a1, Hsd17b, and Cyp19a1, which encode the enzymes catalyzing the synthesis of 17-OHP(4), testosterone, and E(2) respectively. Using a proteomic approach, we identified stathmin (STMN1), as a protein that is overproduced in 17NF ovaries. In its phosphorylated state, STMN1 mediates a cell death signal initiated by tumor necrosis factor alpha (TNF). STMN1 is expressed in GCs and excessive NGF increases its abundance as well as that of its forms phosphorylated at serine (Ser) 16, 25, and 38. TNF synthesis is also increased in 17NF ovaries, and this change is abolished by blocking neurotrophic tyrosine kinase receptors. Inhibiting TNF actions in vivo by administering a soluble TNF receptor prevented the increase in total and phosphorylated STMN1 production, as well as GC apoptosis in NGF-overproducing ovaries. These results indicate that an excess of NGF in the ovary promotes steroidogenesis by enhancing the expression of enzyme genes involved in 17-OHP(4), testosterone, and E(2) synthesis, and causes GC apoptosis by activating a TNF/ STMN1-mediated cell death pathway. PMID- 21646392 TI - Prostaglandin E2-EP4 signaling suppresses adipocyte differentiation in mouse embryonic fibroblasts via an autocrine mechanism. AB - The prostaglandin (PG) receptors EP4 and FP have the potential to exert negative effects on adipogenesis, but the exact contribution of endogenous PG-driven receptor signaling to this process is not fully understood. In this study, we employed an adipocyte differentiation system from mouse embryonic fibroblasts (MEF) and compared the effects of each PG receptor-deficiency on adipocyte differentiation. In wild-type (WT) MEF cells, inhibition of endogenous PG synthesis by indomethacin augmented the differentiation, whereas exogenous PGE2, as well as an FP agonist, reversed the effect of indomethacin. In EP4-deficient cells, basal differentiation was upregulated to the levels in indomethacin treated WT cells, and indomethacin did not further enhance differentiation. Differentiation in FP-deficient cells was equivalent to WT and was still sensitive to indomethacin. PGE2 or indomethacin treatment of WT MEF cells for the first two days was enough to suppress or enhance transcription of the Pparg2 gene as well as the subsequent differentiation, respectively. Differentiation stimuli induced COX-2 gene and protein expression, as well as PGE2 production, in WT MEF cells. These results suggest that PGE2-EP4 signaling suppresses adipocyte differentiation by affecting Pparg2 expression in an autocrine manner and that FP mediated inhibition is not directly involved in adipocyte differentiation in the MEF system. PMID- 21646394 TI - SOCS2 regulates T helper type 2 differentiation and the generation of type 2 allergic responses. AB - The incidence of allergy and asthma in developed countries is on the increase and this trend looks likely to continue. CD4(+) T helper 2 (Th2) cells are major drivers of these diseases and their commitment is controlled by cytokines such as interleukin 4, which are in turn regulated by the suppressor of cytokine signaling (SOCS) proteins. We report that SOCS2(-/-) CD4(+) T cells show markedly enhanced Th2 differentiation. SOCS2(-/-) mice, as well as RAG-1(-/-) mice transferred with SOCS2(-/-) CD4(+) T cells, exhibit elevated type 2 responses after helminth antigen challenge. Moreover, in in vivo models of atopic dermatitis and allergen-induced airway inflammation, SOCS2(-/-) mice show significantly elevated IgE, eosinophilia, type 2 responses, and inflammatory pathology relative to wild-type mice. Finally, after T cell activation, markedly enhanced STAT6 and STAT5 phosphorylation is observed in SOCS2(-/-) T cells, whereas STAT3 phosphorylation is blunted. Thus, we provide the first evidence that SOCS2 plays an important role in regulating Th2 cell expansion and development of the type 2 allergic responses. PMID- 21646395 TI - Destruction of tumor vasculature and abated tumor growth upon VEGF blockade is driven by proapoptotic protein Bim in endothelial cells. AB - For malignant growth, solid cancers must stimulate the formation of new blood vessels by producing vascular endothelial growth factor (VEGF-A), which is required for the survival of tumor-associated vessels. Novel anticancer agents that block VEGF-A signaling trigger endothelial cell (EC) apoptosis and vascular regression preferentially within tumors, but how the ECs die is not understood. In this study, we demonstrate that VEGF-A deprivation, provoked either by drug induced tumor shrinkage or direct VEGF-A blockade, up-regulates the proapoptotic BH3 (Bcl-2 homology 3)-only Bcl-2 family member Bim in ECs. Importantly, the tumor growth inhibitory activity of a VEGF-A antagonist required Bim-induced apoptosis of ECs. These findings thus reveal the mechanism by which VEGF-A blockade induces EC apoptosis and impairs tumor growth. They also indicate that drugs mimicking BH3-only proteins may be exploited to kill tumor cells not only directly but also indirectly by ablating the tumor vasculature. PMID- 21646396 TI - Interaction of the gp120 V1V2 loop with a neighboring gp120 unit shields the HIV envelope trimer against cross-neutralizing antibodies. AB - The HIV-1 envelope trimer adopts a quaternary conformation that effectively shields neutralization-sensitive domains and thus represents a major obstacle for natural and vaccine-elicited antibody responses. By using a structure-function analysis based on a specifically devised mathematical model, we demonstrate in this study that protection from neutralization is enforced by intersubunit contact between the variable loops 1 and 2 (V1V2) and domains of neighboring gp120 subunits in the trimer encompassing the V3 loop. Our data are consistent with an interaction of the V1V2 and V3 loop at the spike apex as proposed by cryoelectron tomography experiments. By defining the orientation of the V1V2 loop within the trimer toward the neighboring gp120 subunit's V3 loop, our data close an important gap in the understanding of the architecture of the trimeric spike. Knowledge on how the V1V2 barrier functions in the context of the trimer to mask conserved epitopes on gp120 may aid future vaccine design. PMID- 21646397 TI - Combined inadequacies of multiple B vitamins amplify colonic Wnt signaling and promote intestinal tumorigenesis in BAT-LacZxApc1638N mice. AB - The Wnt pathway is a pivotal signaling cascade in colorectal carcinogenesis. The purpose of this work is to determine whether depletion of folate and other metabolically related B vitamins induces in vivo activation of intestinal Wnt signaling and whether this occurs in parallel with increased tumorigenesis. A hybrid mouse was created by crossing a Wnt-reporter animal (BAT-LacZ) with a model of colorectal cancer (Apc1638N). A mild depletion of folate and vitamins B2, B6, and B12 was induced over 16 wk, and the control animals in each instance were pair fed a diet containing the basal requirement of these nutrients. The multiplicity of macroscopic tumors and aberrant crypt foci both increased by ~50% in the hybrid mice fed the depletion diet (P<0.05). A 4-fold elevation in Wnt signaling was produced by the depletion diet (P<0.05) and was accompanied by significant changes in the expression of a number of Wnt-related genes in a pattern consistent with its activation. Proliferation and apoptosis of the colonic mucosa both changed in a protransformational direction (P<0.05). In summary, mild depletion of multiple B vitamins produces in vivo activation of colonic Wnt signaling, implicating it as a key pathway by which B-vitamin inadequacies enhance intestinal tumorigenesis. PMID- 21646398 TI - Normobaric hypoxia impairs human cardiac energetics. AB - Hypoxia causes left ventricular dysfunction in the human heart, but the biochemical mechanism is poorly understood. Here, we tested whether short-term normobaric hypoxia leads to changes in cardiac energetics and early cardiac dysfunction. Healthy male volunteers (n=12, age 24 +/- 2 yr) were exposed to normobaric hypoxia in a purpose-built hypoxic chamber. The partial pressure of oxygen during end-tidal expiration (P(ET)o2) was kept between 50 and 60 mmHg, and peripheral oxygen saturation (Sao2) was kept above 80%. Cardiac morphology and function were assessed using magnetic resonance imaging and echocardiography, both before and after 20 h of hypoxic exposure, and high-energy phosphate metabolism [measured as the phosphocreatine (PCr)/ATP ratio] was measured using 31P magnetic resonance spectroscopy. During hypoxia, P(ET)o2 and Sao2 averaged 55 +/- 1 mmHg and 83.6 +/- 0.4%, respectively. Hypoxia caused a 15% reduction in cardiac PCr/ATP (from 2.0 +/- 0.1 to 1.7 +/- 0.1, P<0.01) and reduced diastolic function (measured as E/E', rising from 6.1 +/- 0.4 to 7.5 +/- 0.7, P<0.01). Normobaric hypoxia causes a rapid decrease in high-energy phosphate metabolism in the human cardiac left ventricle, which may lead to a decline in diastolic function. These findings are important in understanding the response of normal individuals to environmental hypoxia, and to situations in which disease reduces cardiac oxygen delivery. PMID- 21646399 TI - Mitochondrial superoxide flashes: metabolic biomarkers of skeletal muscle activity and disease. AB - Mitochondrial superoxide flashes (mSOFs) are stochastic events of quantal mitochondrial superoxide generation. Here, we used flexor digitorum brevis muscle fibers from transgenic mice with muscle-specific expression of a novel mitochondrial-targeted superoxide biosensor (mt-cpYFP) to characterize mSOF activity in skeletal muscle at rest, following intense activity, and under pathological conditions. Results demonstrate that mSOF activity in muscle depended on electron transport chain and adenine nucleotide translocase functionality, but it was independent of cyclophilin-D-mediated mitochondrial permeability transition pore activity. The diverse spatial dimensions of individual mSOF events were found to reflect a complex underlying morphology of the mitochondrial network, as examined by electron microscopy. Muscle activity regulated mSOF activity in a biphasic manner. Specifically, mSOF frequency was significantly increased following brief tetanic stimulation (18.1 +/- 1.6 to 22.3 +/- 2.0 flashes/1000 MUm2.100 s before and after 5 tetani) and markedly decreased (to 7.7 +/- 1.6 flashes/1000 MUm2.100 s) following prolonged tetanic stimulation (40 tetani). A significant temperature-dependent increase in mSOF frequency (11.9 +/- 0.8 and 19.8 +/- 2.6 flashes/1000 MUm2.100 s at 23 degrees C and 37 degrees C) was observed in fibers from RYR1(Y522S/WT) mice, a mouse model of malignant hyperthermia and heat-induced hypermetabolism. Together, these results demonstrate that mSOF activity is a highly sensitive biomarker of mitochondrial respiration and the cellular metabolic state of muscle during physiological activity and pathological oxidative stress PMID- 21646400 TI - New ways to turn on NKT cells. AB - Natural killer T (NKT) cells are CD1d-restricted, lipid antigen-reactive T cells with powerful immunoregulatory potential. The prototypic antigen for NKT cells is a marine sponge-derived glycolipid, alpha-galactosylceramide (alpha-GalCer), but this is not normally encountered in the mammalian environment. Thus, there is great interest in the identification of more physiological stimuli for NKT cells, and numerous studies have shown that NKT cells are capable of responding to a range of microbial lipid-based antigens. Two new studies expand our understanding of environmental NKT cell stimuli, with one showing that CD1d-restricted NKT cell antigens are present within common house dust extract (HDE), whereas the other shows that NKT cells can respond to innate stimuli irrespective of the presence of foreign microbial antigens. Collectively, these two investigations indicate that NKT cells are far more likely to encounter foreign antigens, or innate activating signals, than previously recognized, suggesting a more central role for these cells in the immune system. PMID- 21646401 TI - EB1 promotes microtubule dynamics by recruiting Sentin in Drosophila cells. AB - Highly conserved EB1 family proteins bind to the growing ends of microtubules, recruit multiple cargo proteins, and are critical for making dynamic microtubules in vivo. However, it is unclear how these master regulators of microtubule plus ends promote microtubule dynamics. In this paper, we identify a novel EB1 cargo protein, Sentin. Sentin depletion in Drosophila melanogaster S2 cells, similar to EB1 depletion, resulted in an increase in microtubule pausing and led to the formation of shorter spindles, without displacing EB1 from growing microtubules. We demonstrate that Sentin's association with EB1 was critical for its plus end localization and function. Furthermore, the EB1 phenotype was rescued by expressing an EBN-Sentin fusion protein in which the C-terminal cargo-binding region of EB1 is replaced with Sentin. Knockdown of Sentin attenuated plus end accumulation of Msps (mini spindles), the orthologue of XMAP215 microtubule polymerase. These results indicate that EB1 promotes dynamic microtubule behavior by recruiting the cargo protein Sentin and possibly also a microtubule polymerase to the microtubule tip. PMID- 21646402 TI - Direct regulation of Treslin by cyclin-dependent kinase is essential for the onset of DNA replication. AB - Treslin, a TopBP1-interacting protein, is necessary for deoxyribonucleic acid (DNA) replication in vertebrates. Association between Treslin and TopBP1 requires cyclin-dependent kinase (Cdk) activity in Xenopus laevis egg extracts. We investigated the mechanism and functional importance of Cdk for this interaction using both X. laevis egg extracts and human cells. We found that Treslin also associated with TopBP1 in a Cdk-regulated manner in human cells and that Treslin was phosphorylated within a conserved Cdk consensus target sequence (on S976 in X. laevis and S1000 in humans). Recombinant human Cdk2-cyclin E also phosphorylated this residue of Treslin in vitro very effectively. Moreover, a mutant of Treslin that cannot undergo phosphorylation on this site showed significantly diminished binding to TopBP1. Finally, human cells harboring this mutant were severely deficient in DNA replication. Collectively, these results indicate that Cdk-mediated phosphorylation of Treslin during S phase is necessary for both its effective association with TopBP1 and its ability to promote DNA replication in human cells. PMID- 21646403 TI - Bub1 overexpression induces aneuploidy and tumor formation through Aurora B kinase hyperactivation. AB - High expression of the protein kinase Bub1 has been observed in a variety of human tumors and often correlates with poor clinical prognosis, but its molecular and cellular consequences and role in tumorigenesis are unknown. Here, we demonstrate that overexpression of Bub1 in mice leads to near-diploid aneuploidies and tumor formation. We found that chromosome misalignment and lagging are the primary mitotic errors responsible for the observed aneuploidization. High Bub1 levels resulted in aberrant Bub1 kinase activity and hyperactivation of Aurora B kinase. When Aurora B activity is suppressed, pharmacologically or via BubR1 overexpression, chromosome segregation errors caused by Bub1 overexpression are largely corrected. Importantly, Bub1 transgenic mice overexpressing Bub1 developed various kinds of spontaneous tumors and showed accelerated Myc-induced lymphomagenesis. Our results establish that Bub1 has oncogenic properties and suggest that Aurora B is a critical target through which overexpressed Bub1 drives aneuploidization and tumorigenesis. PMID- 21646404 TI - SLAIN2 links microtubule plus end-tracking proteins and controls microtubule growth in interphase. AB - The ends of growing microtubules (MTs) accumulate a set of diverse factors known as MT plus end-tracking proteins (+TIPs), which control microtubule dynamics and organization. In this paper, we identify SLAIN2 as a key component of +TIP interaction networks. We showed that the C-terminal part of SLAIN2 bound to end binding proteins (EBs), cytoplasmic linker proteins (CLIPs), and CLIP-associated proteins and characterized in detail the interaction of SLAIN2 with EB1 and CLIP 170. Furthermore, we found that the N-terminal part of SLAIN2 interacted with ch TOG, the mammalian homologue of the MT polymerase XMAP215. Through its multiple interactions, SLAIN2 enhanced ch-TOG accumulation at MT plus ends and, as a consequence, strongly stimulated processive MT polymerization in interphase cells. Depletion or disruption of the SLAIN2-ch-TOG complex led to disorganization of the radial MT array. During mitosis, SLAIN2 became highly phosphorylated, and its interaction with EBs and ch-TOG was inhibited. Our study provides new insights into the molecular mechanisms underlying cell cycle specific regulation of MT polymerization and the organization of the MT network. PMID- 21646405 TI - The reduction in human motoneurone responsiveness during muscle fatigue is not prevented by increased muscle spindle discharge. AB - Motoneurone excitability is rapidly and profoundly reduced during a sustained maximal voluntary contraction (MVC) when tested in the transient silent period which follows transcranial magnetic stimulation (TMS) of the motor cortex. One possible cause of this reduction in excitability is a fatigue-induced withdrawal of excitatory input to motoneurones from muscle spindle afferents. We aimed to test if muscle spindle input produced by tendon vibration would ameliorate suppression of the cervicomedullary motor-evoked potential (CMEP) in the silent period during a sustained MVC. Seven subjects performed a 2 min MVC of the elbow flexors. Stimulation of the corticospinal tract at the level of the mastoids was preceded 100 ms earlier by TMS. These stimulus pairs were delivered every 10 s during the 2 min MVC. Stimulus pairs at 30, 50, 70, 90 and 110 s were delivered while vibration (-80 Hz) was applied to the distal tendon of biceps. On a separate day, the protocol was repeated with both stimuli delivered to the motor cortex. The CMEP in the silent period decreased rapidly with fatigue (to -9% of control) and was not affected by tendon vibration (P = 0.766). The motor-evoked potential in the silent period also declined rapidly (to -5% of control) and was similarly unaffected by tendon vibration (P = 0.075). These data suggest motoneurone disfacilitation due to a fatigue-related decrease of muscle spindle discharge does not contribute significantly to the profound suppression of motoneurone excitability during the silent period. Therefore, a change to intrinsic motoneurone properties caused by repetitive discharge is most probably responsible. PMID- 21646406 TI - Diversity of neuropsin (KLK8)-dependent synaptic associativity in the hippocampal pyramidal neuron. AB - Hippocampal early (E-) long-term potentiation (LTP) and long-term depression (LTD) elicited by a weak stimulus normally fades within 90 min. Late (L-) LTP and LTD elicited by strong stimuli continue for >180 min and require new protein synthesis to persist. If a strong tetanus is applied once to synaptic inputs, even a weak tetanus applied to another synaptic input can evoke persistent LTP. A synaptic tag is hypothesized to enable the capture of newly synthesized synaptic molecules. This process, referred to as synaptic tagging, is found between not only the same processes (i.e. E- and L-LTP; E- and L-LTD) but also between different processes (i.e. E-LTP and L-LTD; E-LTD and L-LTP) induced at two independent synaptic inputs (cross-tagging). However, the mechanisms of synaptic tag setting remain unclear. In our previous study, we found that synaptic associativity in the hippocampal Schaffer collateral pathway depended on neuropsin (kallikrein-related peptidase 8 or KLK8), a plasticity-related extracellular protease. In the present study, we investigated how neuropsin participates in synaptic tagging and cross-tagging. We report that neuropsin is involved in synaptic tagging during LTP at basal and apical dendritic inputs. Moreover, neuropsin is involved in synaptic tagging and cross-tagging during LTP at apical dendritic inputs via integrin beta1 and calcium/calmodulin-dependent protein kinase II signalling. Thus, neuropsin is a candidate molecule for the LTP specific tag setting and regulates the transformation of E- to L-LTP during both synaptic tagging and cross-tagging. PMID- 21646407 TI - On the contribution of group III and IV muscle afferents to the circulatory response to rhythmic exercise in humans. AB - We investigated the role of skeletal muscle afferent feedback in circulatory control during rhythmic exercise in humans. Nine healthy males performed single leg knee-extensor exercise (15/30/45 watts, 3 min each) under both control conditions (Ctrl) and with lumbar intrathecal fentanyl impairing MU-opioid receptor-sensitive muscle afferents. Cardiac output and femoral blood flow were determined, and femoral arterial/venous blood samples were collected during the final minute of each workload. To rule out cephalad migration of fentanyl to the brainstem,we documented unchanged resting ventilatory responses to different levels of hypercapnia. There were no haemodynamic differences between conditions at rest. However, during exercise cardiac output was -2 % lower with fentanyl blockade compared to control (P < 0.05), secondary to a 6% and 13% reduction in heart rate and stroke volume, respectively. Throughout exercise mean arterial pressure (MAP) was reduced by 7% (P < 0.01) which is likely to have contributed to the 15% fall in femoral blood flow. However, MAP was not completely responsible for this peripheral haemodynamic change as vascular conductance was also attenuated (-9%). Evidence of increasing noradrenaline spillover (P = 0.09) implicated an elevation in sympathetic outflow in this response. The attenuated femoral blood flow during exercise with fentanyl was associated with a 17%reduction in leg O2 delivery (P < 0.01) and a concomitant rise in the arteriovenous O2 difference (4-9%), but leg O2 consumption remained 7-13% lower than control (P < 0.05). Our findings reveal an essential contribution of continuous muscle afferent feedback to ensure the appropriate haemodynamic and ultimately metabolic response to rhythmic exercise in humans PMID- 21646408 TI - Maternal dexamethasone and EEG hyperactivity in preterm fetal sheep. AB - Maternal treatment with synthetic corticosteroids such as dexamethasone (DEX)significantly reduces neonatal morbidity and mortality, but its effects on the fetal brain remain unclear. In this study we evaluated the effects of DEX on EEG activity in preterm fetal sheep. Ewes at 103 days gestation received two intramuscular injections of DEX (12 mg, n = 8) or saline vehicle (n = 7) 24 h apart. Fetal EEG activity was recorded from 6 h before until 120 h after the first injection (DEX-1). DEX-1 was associated with a marked transient rise in total EEG power, maximal at 12 h (P < 0.001), with a relative increase in delta and reduced theta, alpha and beta activity, resolving by 24 h. Continuous EEG records showed a shift to larger but less frequent transient waveforms (P < 0.001). Unexpectedly, evolving epileptiform activity, consistent with electrographic and clinical seizures, developed from 178 +/- 44 min after DEX 1.Similar but smaller changes were seen after the second injection. Following the injections, total power returned to control values, but the proportion of alpha activity progressively increased vs. controls (P < 0.001), with reduced interburst interval duration and number (P < 0.001). No histological neural injury or microglial activation was seen. In summary, exposure to maternal dexamethasone was associated with dramatic, evolving low-frequency hyperactivity on fetal cortical EEG recordings, followed by sustained changes consistent with maturation of fetal sleep architecture. We postulate that these effects may contribute to improved neonatal outcomes. PMID- 21646409 TI - Distinct cellular properties of identified dopaminergic and GABAergic neurons in the mouse ventral tegmental area. AB - The midbrain ventral tegmental area (VTA) contains neurons largely with either a dopaminergic (DAergic) or GABAergic phenotype. Physiological and pharmacological properties of DAergic neurons have been determined using tyrosine hydroxylase (TH) immunohistochemistry but many properties overlap with non-DAergic neurons presumed to be GABAergic. This study examined properties of GABAergic neurons, non-GABAergic neurons and TH-immunopositive neurons in VTA of GAD67-GFP knock-in mice. Ninety-eight per cent of VTA neurons were either GAD-GFP or TH positive,with the latter being five times more abundant. During cell-attached patch-clamp recordings, GAD-GFP neurons fired brief action potentials that could be completely distinguished from those of non-GFP neurons. Pharmacologically, the MU-opioid agonist DAMGO inhibited firing of action potentials in 92% of GAD-GFP neurons but had no effect in non-GFP neurons. By contrast, dopamine invariably inhibited action potentials in non-GFP neurons but only did so in 8% of GAD-GFP neurons. During whole-cell recordings, the narrower width of action potential in GAD-GFP neurons was also evident but there was considerable overlap with non-GFP neurons. GAD-GFP neurons invariably failed to exhibit the potassium-mediated slow depolarizing potential during injection of positive current that was present in all non-GFP neurons. Under voltage-clamp the cationic current, I(h), was found in both types of neurons with considerable overlap in both amplitude and kinetics. These distinct cellular properties may thus be used to confidently discriminate GABAergic and DAergic neurons in VTA during in vitro electrophysiological recordings. PMID- 21646410 TI - Targeting the Nrf2-Keap1 antioxidant defence pathway for neurovascular protection in stroke. AB - Endogenous defence mechanisms by which the brain protects itself against noxious stimuli and recovers from ischaemic damage are a key target of stroke research. The loss of viable brain tissue in the ischaemic core region after stroke is associated with damage to the surrounding area known as the penumbra. Activation of the redox-sensitive transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2) plays a pivotal role in the cellular defence against oxidative stress via transcriptional upregulation of phase II defence enzymes and antioxidant stress proteins. Although recent evidence implicates Nrf2 in neuroprotection, it is not known whether activation of this pathway within the neurovascular unit protects the brain against blood-brain barrier breakdown and cerebrovascular inflammation. Targeting the neurovascular unit should provide novel insights for effective treatment strategies and facilitate translation of experimental findings into clinical therapy. This review focuses on the cytoprotective role of Nrf2 in stroke and examines the evidence that the Nrf2 Keap1 defence pathway may serve as a therapeutic target for neurovascular protection. PMID- 21646411 TI - Two distinct vesicle pools for depolarization-induced exocytosis in somata of dorsal root ganglion neurons. AB - The somata of dorsal root ganglion (DRG) neurons release neurotransmitters and neuropeptides. In addition to the conventional Ca2+-dependent secretion (CDS), Ca2+-independent but voltage-dependent secretion (CIVDS) also occurs in the somata of DRG neurons. Electrical stimulation induces both CDS and CIVDS, which differ in size and are coupled with different types of endocytosis contributed by CIVDS and CDS, respectively. However, it is unclear whether they use a common vesicle pool, so we investigated the relationship between the vesicle pools of CDS and CIVDS. Membrane capacitance recording and photolysis of a caged-Ca2+ compound showed that, in low external Ca2+ solutions, the depolarization-induced exocytosis contained two (fast and slow) phases, which were contributed by CIVDS and CDS, respectively. Depletion of the CDS readily releasable pool using photolysis did not affect the CIVDS. When the CIVDS and CDS vesicle pools were depleted by electrical stimulation, the pools had different sizes. Their kinetics of exocytosis-coupled endocytosis were also different. Thus, CIVDS and CDS used different vesicle pools in DRG neurons. PMID- 21646413 TI - Retraction: Exercise induces interleukin-8 expression in human skeletal muscle. PMID- 21646412 TI - Cannabinoid receptor agonists potentiate action potential-independent release of GABA in the dentate gyrus through a CB1 receptor-independent mechanism. AB - We report a novel excitatory effect of cannabinoid agonists on action potential independent GABAergic transmission in the rat dentate gyrus. Specifically, we find that both WIN55,212-2 and anandamide increase the frequency of miniature IPSCs (mIPSCs)recorded from hilar mossy cells without altering event amplitude, area, rise time, or decay. The effect of WIN55,212-2 on mIPSCs is insensitive to AM251 and preserved in CB1 -/- animals,indicating that it does not depend on activation of CB1 receptors. It is also insensitive to AM630 and unaffected by capsazepine suggesting that neither CB2 nor TRPV1 receptors are involved. Further, it is blocked by pre-incubation in suramin and by a selective protein kinase A inhibitor (H-89), and is mimicked (and occluded) by bath application of forskolin. Similar CB1 receptor-independent facilitation of exocytosis is not apparent when recording evoked IPSCs in the presence of AM251, suggesting that the exocytotic mechanism that produces WIN55,212-2 sensitive mIPSCs is distinct from that which produces CB1 sensitive and action potential-dependent release. Despite clear independence from action potentials, WIN55,212-2 mediated facilitation of mIPSCs requires calcium, and yet is insensitive to chelation of calcium in the postsynaptic cell. Finally, we demonstrate that both bath application of 2-arachidonoylglycerol(2-AG) and depolarization-induced release of endogenous cannabinoids have minimal effect on mIPSC frequency. Cumulatively, our results indicate that cannabinoid ligands can selectively facilitate action potential-independent exocytosis of GABA in the rat dentate gyrus, and further emphasize that this new cannabinoid sensitive signalling system is distinct from previously described CB1 receptor-dependent systems in numerous respects. PMID- 21646414 TI - Automated measurement of zebrafish larval movement. AB - The zebrafish is a powerful vertebrate model that is readily amenable to genetic, pharmacological and environmental manipulations to elucidate the molecular and cellular basis of movement and behaviour. We report software enabling automated analysis of zebrafish movement from video recordings captured with cameras ranging from a basic camcorder to more specialized equipment. The software, which is provided as open-source MATLAB functions, can be freely modified and distributed, and is compatible with multiwell plates under a wide range of experimental conditions. Automated measurement of zebrafish movement using this technique will be useful for multiple applications in neuroscience, pharmacology and neuropsychiatry. PMID- 21646415 TI - IgG4 multiorgan lymphoproliferative syndrome as a differential diagnosis of primary Sjogren's syndrome in men? PMID- 21646416 TI - Lumbar disc degeneration and genetic factors are the main risk factors for low back pain in women: the UK Twin Spine Study. AB - OBJECTIVE: Low back pain (LBP) is a common musculoskeletal disorder, but it is still unclear which individuals develop it. The authors examined the contribution of genetic factors, lumbar disc degeneration (LDD) and other risk factors in a female sample of the general population. MATERIAL AND METHODS: A cross-sectional study was conducted among 2256 women (371 and 698 monozygotic and dizygotic twin pairs and 29 sibling pairs and 60 singletons) with a mean age of 50 years (18 84). A self-reported validated questionnaire was used to collect back pain data. Risk factors including body weight, smoking, occupation, physical exercise and MRI assessed LDD were measured. Data analysis included logistic regression and variance decomposition. RESULTS: The major factors associated with LBP included genetic background, with OR approximately 6 if the monozygotic co-twin had LBP, or 2.2 if she was a dizygotic co-twin. In addition, LDD and overweight were highly significantly (p<0.001) associated with non-specific LBP. The single most important risk factor was the amount of LDD. After adjustment for other risk factors, the individuals who exhibited advanced LDD (90% vs 10%) had 3.2 higher odds of manifesting LBP. The data also showed a significant (p<0.001) genetic correlation between the LBP and LDD measurements, suggesting that approximately 11-13% of the genetic effects are shared by LDD and LBP. CONCLUSIONS: The main risk factors for reported episodes of severe and disabling LBP in UK women include the degree of LDD as assessed by MRI, being overweight and genetic heritability. PMID- 21646418 TI - Evaluation of a telephone speech-enhancement algorithm among older adults with hearing loss. AB - PURPOSE: In this study, the authors evaluated a processing algorithm aimed at improving speech recognition via the telephone among older adults with sensorineural hearing loss (SNHL). METHOD: Thirty older adults with SNHL participated. Speech recognition was measured in quiet using the Modified Rhyme Test (MRT; Kreul et al., 1968) and the Speech Perception in Noise (SPIN; Bilger et al., 1984) sentences, and in noise using the Quick Speech in Noise (QSIN; Killion et al., 2004) test. Each test was presented via the telephone with and without processing. RESULTS: Significant improvements in recognition performance due to processing were observed for the SPIN and QSIN. The improvement on the QSIN was significantly greater than on the MRT and SPIN, likely because the MRT and SPIN sentences were presented in quiet, whereas the QSIN was presented in noise. Significant improvements in recognition performance were observed for both an offline version and a real-time version of the algorithm relative to the unprocessed condition, although no difference was noted between the 2 versions. CONCLUSIONS: Results indicate that preprocessing the acoustic signal is a viable method of improving speech recognition via the telephone. The algorithm has the potential to benefit older adults with SNHL who struggle to communicate via the telephone with or without hearing aids. PMID- 21646417 TI - Risk factors for medial meniscal pathology on knee MRI in older US adults: a multicentre prospective cohort study. AB - OBJECTIVES: Meniscal pathology in which the aetiology is often unclear is a frequent finding on knee MRI. This study investigates potential risk factors for medial meniscal lesions or extrusion in middle-aged and elderly persons. METHODS: Prospective cohort study using population-based subjects from Birmingham, Alabama and Iowa City, Iowa, USA (the Multicenter Osteoarthritis Study). 644 men and women aged 50-79 years with or at high risk of knee osteoarthritis (Kellgren and Lawrence grade 0-2) but with normal medial meniscal status at baseline were studied. Paired baseline and 30-month 1.0 T knee MRI were scored for meniscal lesions and extrusion (pathology) and the following systemic, knee-specific and compartment-specific potential risk factors were evaluated: age, sex, body mass index, bony enlargement of finger joints, knee trauma, leg-length inequality and knee alignment. RESULTS: Of 791 knees, 77 (9.7%) had medial meniscal pathology at 30 months follow-up. 61 of the 77 (81%) had no report of trauma during follow-up. Including all potential risk factors in the multivariable model, the adjusted OR for medial meniscal pathology was 4.14 (95% CI 2.06 to 8.31) for knee trauma during follow-up, 1.64 (1.00 to 2.70) for five or more bony enlargements of finger joints (vs <= 4) and 2.00 (1.18 to 3.40) for varus alignment (vs not varus) at baseline examination. Obesity was a risk factor for the development of meniscal extrusion, OR 3.04 (1.04 to 8.93) but not for meniscal lesions, OR 1.15 (0.52 to 2.54). CONCLUSIONS: Apart from knee trauma, possible generalised osteoarthritis, expressed as multiple bony enlargements of finger joints, varus alignment and obesity are risk factors for medial meniscal pathology. PMID- 21646419 TI - Sustained attention in children with primary language impairment: a meta analysis. AB - PURPOSE: This study provides a meta-analysis of the difference between children with primary or specific language impairment (LI) and their typically developing peers on tasks of sustained attention. The meta-analysis seeks to determine whether children with LI demonstrate subclinical deficits in sustained attention and, if so, under what conditions. METHOD: Articles that reported empirical data from the performance of children with LI, in comparison to typically developing peers, on a task assessing sustained attention were considered for inclusion. Twenty-eight effect sizes were included in the meta-analysis. Two moderator analyses addressed the effects of stimulus modality and attention deficit/hypereactivity disorder exclusion. In addition, reaction time outcomes and the effects of task variables were summarized qualitatively. RESULTS: The meta-analysis supports the existence of sustained attention deficits in children with LI in both auditory and visual modalities, as demonstrated by reduced accuracy compared with typically developing peers. Larger effect sizes are found in tasks that use auditory-linguistic stimuli than in studies that use visual stimuli. CONCLUSIONS: Future research should consider the role that sustained attention weaknesses play in LI as well as the implications for clinical and research assessment tasks. Methodological recommendations are summarized. PMID- 21646420 TI - Enhancing speech discrimination through stimulus repetition. AB - PURPOSE: To evaluate the effects of sequential and alternating repetition on speech-sound discrimination. METHOD: Typically hearing adults' discrimination of 3 pairs of speech-sound contrasts was assessed at 3 signal-to-noise ratios using the change/no-change procedure. On change trials, the standard and comparison stimuli differ; on no-change trials, they are identical. Listeners were presented with 5 repetition conditions: 2 and 4 sequential repetitions of the standard followed by sequential repetitions of the comparison; 2 and 4 alternating presentations of the standard and comparison; and 1 repetition of the standard and comparison. RESULTS: Both sequential and alternating repetition improved discrimination of the fricative and liquid contrasts, but neither was clearly superior to the other across the conditions. CONCLUSIONS: The results support previous findings that increasing the number of fricative and liquid stimulus presentations improves discriminability and extends the findings to natural speech stimuli. Further, the effect of repetition is robust: Both sequential and alternating repetitions improve speech-sound discrimination, and few differences emerge between the two types of stimulus repetitions. The results have implications for evaluating the strength of the internal representation of speech stimuli in clinical populations believed to have a core deficit in phonological encoding, such as children with hearing loss. PMID- 21646421 TI - Classifications of vocalic segments from articulatory kinematics: healthy controls and speakers with dysarthria. AB - PURPOSE: In this study, the authors classified vocalic segments produced by control speakers (C) and speakers with dysarthria due to amyotrophic lateral sclerosis (ALS) or Parkinson's disease (PD); classification was based on movement measures. The researchers asked the following questions: (a) Can vowels be classified on the basis of selected measures of articulatory motions? and (b) Can classification models that are constructed from control productions classify vowels produced by speakers with dysarthria that is related to ALS and PD? METHOD: Nineteen C, 7 PD, and 8 ALS speakers participated in this study. The severity of dysarthria varied across individuals and between the 2 disorder groups. The stimuli were 6 vowels produced in 10 words embedded into sentences read at a comfortable reading rate. Movement data were collected using the x-ray microbeam. Movement measures included distances traveled, durations, and average speeds of vowel-related movement strokes. Vowels and words were classified by linear discriminant analysis with measures of articulatory motion as input variables. RESULTS: The study showed that vocalic segments could be classified using articulatory movement characteristics with up to 80% accuracy. The classification accuracy of the movement-based models depended largely on the number of articulators involved and, to a lesser extent, on the movement measure (e.g., distance, duration, speed). Classification of PD vowels was similar to that of the C group, suggesting a simple scaling of gestures as an explanation of the movement deficit in this disease. Classification performance for ALS vowels appeared to be different from that of C and PD productions. CONCLUSION: Classification of vowels was possible on the basis of their articulatory motions. ALS vowels appeared categorically different from those of C and PD speakers. PMID- 21646422 TI - Taxometric analyses of specific language impairment in 6-year-old children. AB - PURPOSE: To determine whether language scores at age 6 years suggest that specific language impairments (SLIs) distribute in a categorical or in a dimensional fashion. METHOD: A taxometric analysis of language scores from 601 six-year-old children who were free of neonatal risk factors was performed. From among 4 candidate indicators of SLI, 2 were eligible for the mean above minus below a cut (MAMBAC) procedure (Meehl & Yonce, 1994): number of different words (NDW) produced in a language sample and average percentage phonemes repeated correctly in 3- and 4-syllable nonwords (3-4 PPC). Graphs of these MAMBAC variables were inspected to determine whether they patterned in a manner suggesting the presence of a discrete category, having either a central peak or a steep curve peaking near the final interval. RESULTS: MAMBAC curves for NDW and 3 4 PPC did not suggest the presence of a taxon, and results did not vary during consistency checks in which MAMBAC parameters and cutoff values for language deficits were manipulated. CONCLUSIONS: Results suggest that for these measures, children with specific language deficits are those falling at the lower end of a continuous distribution of language skills rather than a qualitatively distinct group. PMID- 21646423 TI - Age-related benefits of digital noise reduction for short-term word learning in children with hearing loss. AB - PURPOSE: To determine the rate of word learning for children with hearing loss (HL) in quiet and in noise compared to normal-hearing (NH) peers. The effects of digital noise reduction (DNR) were examined for children with HL. METHOD: Forty one children with NH and 26 children with HL were grouped by age (8-9 years and 11-12 years). The children learned novel words associated with novel objects through a process of trial and error. Functions relating performance across trials were calculated for each child in each listening condition and were compared. RESULTS: Significant effects were observed for age (older > younger) in the children with NH and listening condition (quiet > noise) in the children with HL. Significant effects of hearing status were also observed across groups (NH > HL), indicating that the children with HL required more trials to learn the new words. However, word learning improved significantly in noise with the use of DNR for the older but not for the younger children with HL. Hearing aid history and signal-to-noise ratio did not contribute to performance. CONCLUSION: Word learning was significantly reduced in younger children, in noise, and in the presence of hearing loss. Age-related benefits of DNR were apparent for children over 10 years of age. PMID- 21646424 TI - Histone H3 lysine 4 hypermethylation prevents aberrant nucleosome remodeling at the PHO5 promoter. AB - Recent studies have highlighted the histone H3K4 methylation (H3K4me)-dependent transcriptional repression in Saccharomyces cerevisiae; however, the underlying mechanism remains inexplicit. Here, we report that H3K4me inhibits the basal PHO5 transcription under high-phosphate conditions by suppressing nucleosome disassembly at the promoter. We found that derepression of the PHO5 promoter by SET1 deletion resulted in a labile chromatin structure, allowing more binding of RNA polymerase II (Pol II) but not the transactivators Pho2 and Pho4. We further showed that Pho23 and Cti6, two plant homeodomain (PHD)-containing proteins, cooperatively anchored the large Rpd3 (Rpd3L) complex to the H3K4-methylated PHO5 promoter. The deacetylation activity of Rpd3 on histone H3 was required for the function of Set1 at the PHO5 promoter. Taken together, our data suggest that Set1 mediated H3K4me suppresses nucleosome remodeling at the PHO5 promoter so as to reduce basal transcription of PHO5 under repressive conditions. We propose that the restriction of aberrant nucleosome remodeling contributes to strict control of gene transcription by the transactivators. PMID- 21646425 TI - miR-29a and miR-29b contribute to pancreatic beta-cell-specific silencing of monocarboxylate transporter 1 (Mct1). AB - In pancreatic beta cells, elevated glucose concentrations stimulate mitochondrial oxidative metabolism to raise intracellular ATP/ADP levels, prompting insulin secretion. Unusually low levels of expression of genes encoding the plasma membrane monocarboxylate transporter, MCT1 (SLC16A1), as well as lactate dehydrogenase A (LDHA) ensure that glucose-derived pyruvate is efficiently metabolized by mitochondria, while exogenous lactate or pyruvate is unable to stimulate metabolism and hence insulin secretion inappropriately. We show here that whereas DNA methylation at the Mct1 promoter is unlikely to be involved in cell-type-specific transcriptional repression, three microRNAs (miRNAs), miR-29a, miR-29b, and miR-124, selectively target both human and mouse MCT1 3' untranslated regions. Mutation of the cognate miR-29 or miR-124 binding sites abolishes the effects of the corresponding miRNAs, demonstrating a direct action of these miRNAs on the MCT1 message. However, despite reports of its expression in the mouse beta-cell line MIN6, miR-124 was not detectably expressed in mature mouse islets. In contrast, the three isoforms of miR-29 are highly expressed and enriched in mouse islets. We show that inhibition of miR-29a in primary mouse islets increases Mct1 mRNA levels, demonstrating that miR-29 isoforms contribute to the beta-cell-specific silencing of the MCT1 transporter and may thus affect insulin release. PMID- 21646426 TI - The glucocorticoid receptor and the coregulator Brm selectively modulate each other's occupancy and activity in a gene-specific manner. AB - The diverse transcriptional patterns that distinguish metazoan cells are specified by multifactor regulatory complexes containing distinct combinations of factors that assemble at genomic response elements. To investigate combinatorial control, we examined a set of glucocorticoid receptor (GR)-regulated genes bearing nearby regulatory complexes that include both GR and the coregulator Brm, an ATPase subunit of the Swi/Snf chromatin remodeler. We analyzed how GR and Brm affect each other's occupancy and activity by utilizing glucocorticoid treatment and Brm knockdown to modulate GR-mediated transcriptional regulation and Brm mediated chromatin remodeling, respectively. GR occupancy and activity were altered differentially by Brm knockdown at specific activated and repressed primary GR target genes. Brm knockdown decreased GR occupancy at activated Brm dependent genes, whereas we identified two classes of repressed genes, at which Brm knockdown either increased or decreased GR occupancy. Glucocorticoid treatment increased both Brm occupancy and chromatin accessibility at Brm dependent and Brm-independent GR-regulated genes. However, chromatin remodeling activity decreased after Brm knockdown only at genes with Brm-dependent transcription. Our study revealed multiple distinct patterns of GR and Brm interdependence. Thus, monitoring as few as two factors within regulatory complexes is sufficient to reveal functionally distinct assemblies, providing an analytical method for gaining insights into combinatorial regulation. PMID- 21646427 TI - Importin alpha-mediated nuclear import of cytoplasmic poly(A) binding protein occurs as a direct consequence of cytoplasmic mRNA depletion. AB - Recent studies have found the cytoplasmic poly(A) binding protein (PABPC) to have opposing effects on gene expression when concentrated in the cytoplasm versus in the nucleus. PABPC is predominantly cytoplasmic at steady state, where it enhances protein synthesis through simultaneous interactions with mRNA and translation factors. However, it accumulates dramatically within the nucleus in response to various pathogenic and nonpathogenic stresses, leading to an inhibition of mRNA export. The molecular events that trigger relocalization of PABPC and the mechanisms by which it translocates into the nucleus to block gene expression are not understood. Here, we reveal an RNA-based mechanism of retaining PABPC in the cytoplasm. Expression either of viral proteins that promote mRNA turnover or of a cytoplasmic deadenylase drives nuclear relocalization of PABPC in a manner dependent on the PABPC RNA recognition motifs (RRMs). Using multiple independent binding sites within its RRMs, PABPC interacts with importin alpha, a component of the classical import pathway. Finally, we demonstrate that the direct association of PABPC with importin alpha is antagonized by the presence of poly(A) RNA, supporting a model in which RNA binding masks nuclear import signals within the PABPC RRMs, thereby ensuring efficient cytoplasmic retention of this protein in normal cells. These findings further suggest that cells must carefully calibrate the ratio of PABPC to mRNA, as events that offset this balance can dramatically influence gene expression. PMID- 21646429 TI - Atomic force microscopy in imaging of viruses and virus-infected cells. AB - Atomic force microscopy (AFM) can visualize almost everything pertinent to structural virology and at resolutions that approach those for electron microscopy (EM). Membranes have been identified, RNA and DNA have been visualized, and large protein assemblies have been resolved into component substructures. Capsids of icosahedral viruses and the icosahedral capsids of enveloped viruses have been seen at high resolution, in some cases sufficiently high to deduce the arrangement of proteins in the capsomeres as well as the triangulation number (T). Viruses have been recorded budding from infected cells and suffering the consequences of a variety of stresses. Mutant viruses have been examined and phenotypes described. Unusual structural features have appeared, and the unexpectedly great amount of structural nonconformity within populations of particles has been documented. Samples may be imaged in air or in fluids (including culture medium or buffer), in situ on cell surfaces, or after histological procedures. AFM is nonintrusive and nondestructive, and it can be applied to soft biological samples, particularly when the tapping mode is employed. In principle, only a single cell or virion need be imaged to learn of its structure, though normally images of as many as is practical are collected. While lateral resolution, limited by the width of the cantilever tip, is a few nanometers, height resolution is exceptional, at approximately 0.5 nm. AFM produces three-dimensional, topological images that accurately depict the surface features of the virus or cell under study. The images resemble common light photographic images and require little interpretation. The structures of viruses observed by AFM are consistent with models derived by X-ray crystallography and cryo-EM. PMID- 21646430 TI - cis-antisense RNA, another level of gene regulation in bacteria. AB - A substantial amount of antisense transcription is a hallmark of gene expression in eukaryotes. However, antisense transcription was first demonstrated in bacteria almost 50 years ago. The transcriptomes of bacteria as different as Helicobacter pylori, Bacillus subtilis, Escherichia coli, Synechocystis sp. strain PCC6803, Mycoplasma pneumoniae, Sinorhizobium meliloti, Geobacter sulfurreducens, Vibrio cholerae, Chlamydia trachomatis, Pseudomonas syringae, and Staphylococcus aureus have now been reported to contain antisense RNA (asRNA) transcripts for a high percentage of genes. Bacterial asRNAs share functional similarities with trans-acting regulatory RNAs, but in addition, they use their own distinct mechanisms. Among their confirmed functional roles are transcription termination, codegradation, control of translation, transcriptional interference, and enhanced stability of their respective target transcripts. Here, we review recent publications indicating that asRNAs occur as frequently in simple unicellular bacteria as they do in higher organisms, and we provide a comprehensive overview of the experimentally confirmed characteristics of asRNA actions and intimately linked quantitative aspects. Emerging functional data suggest that asRNAs in bacteria mediate a plethora of effects and are involved in far more processes than were previously anticipated. Thus, the functional impact of asRNAs should be considered when developing new strategies against pathogenic bacteria and when optimizing bacterial strains for biotechnology. PMID- 21646428 TI - Regulatory circuitry governing fungal development, drug resistance, and disease. AB - Pathogenic fungi have become a leading cause of human mortality due to the increasing frequency of fungal infections in immunocompromised populations and the limited armamentarium of clinically useful antifungal drugs. Candida albicans, Cryptococcus neoformans, and Aspergillus fumigatus are the leading causes of opportunistic fungal infections. In these diverse pathogenic fungi, complex signal transduction cascades are critical for sensing environmental changes and mediating appropriate cellular responses. For C. albicans, several environmental cues regulate a morphogenetic switch from yeast to filamentous growth, a reversible transition important for virulence. Many of the signaling cascades regulating morphogenesis are also required for cells to adapt and survive the cellular stresses imposed by antifungal drugs. Many of these signaling networks are conserved in C. neoformans and A. fumigatus, which undergo distinct morphogenetic programs during specific phases of their life cycles. Furthermore, the key mechanisms of fungal drug resistance, including alterations of the drug target, overexpression of drug efflux transporters, and alteration of cellular stress responses, are conserved between these species. This review focuses on the circuitry regulating fungal morphogenesis and drug resistance and the impact of these pathways on virulence. Although the three human-pathogenic fungi highlighted in this review are those most frequently encountered in the clinic, they represent a minute fraction of fungal diversity. Exploration of the conservation and divergence of core signal transduction pathways across C. albicans, C. neoformans, and A. fumigatus provides a foundation for the study of a broader diversity of pathogenic fungi and a platform for the development of new therapeutic strategies for fungal disease. PMID- 21646431 TI - Nucleosome positioning in Saccharomyces cerevisiae. AB - The DNA of eukaryotic cells is spooled around large histone protein complexes, forming nucleosomes that make up the basis for a high-order packaging structure called chromatin. Compared to naked DNA, nucleosomal DNA is less accessible to regulatory proteins and regulatory processes. The exact positions of nucleosomes therefore influence several cellular processes, including gene expression, chromosome segregation, recombination, replication, and DNA repair. Here, we review recent technological advances enabling the genome-wide mapping of nucleosome positions in the model eukaryote Saccharomyces cerevisiae. We discuss the various parameters that determine nucleosome positioning in vivo, including cis factors like AT content, variable tandem repeats, and poly(dA:dT) tracts that function as chromatin barriers and trans factors such as chromatin remodeling complexes, transcription factors, histone-modifying enzymes, and RNA polymerases. In the last section, we review the biological role of chromatin in gene transcription, the evolution of gene regulation, and epigenetic phenomena. PMID- 21646434 TI - Intra-articular findings in primary and revision anterior cruciate ligament reconstruction surgery: a comparison of the MOON and MARS study groups. AB - BACKGROUND: At the time of anterior cruciate ligament (ACL) reconstruction, there are usually concurrent meniscal and articular cartilage injuries. It is unclear if there is a significant difference between intra-articular injuries at the time of a primary ACL reconstruction compared with revision ACL reconstruction. PURPOSE: To compare the meniscal and articular cartilage injuries found at the time of primary and revision ACL reconstruction surgery and to determine associations between primary and revision surgery and specific intra-articular findings. STUDY DESIGN: Cohort study (prevalence); Level of evidence, 2. METHODS: Primary and revision ACL surgeries were identified from the Multicenter Orthopedic Outcomes Network (MOON) and Multicenter ACL Revision Study (MARS) study groups, respectively, from January 1, 2007 to November 1, 2008. Demographic data on individual patients were analyzed including age, body mass index (BMI), and gender. Intra-articular findings including the presence of medial or lateral meniscal tears and chondral damage to articular surfaces were analyzed for each patient. Comparisons of intra-articular findings at the time of surgery for the 2 groups were analyzed. Chondral damage in the medial and lateral compartments was analyzed considering previous meniscal tear as a possible confounder. RESULTS: There were 508 patients undergoing primary ACL reconstruction and 281 patients undergoing revision ACL reconstruction who were identified for inclusion. There were no differences in the mean age, BMI, and gender in the 2 study groups. There was a decreased odds ratio (OR) of new untreated lateral meniscal tears (OR, 0.54; P < .01) but not of medial meniscal tears (OR, 0.86; P = .39) in revision compared with primary ACL reconstruction. There was an increased OR of Outerbridge grade 3 and 4 articular cartilage injury in revision compared with primary ACL reconstruction in the lateral compartment (OR, 1.73; P = .04) and in the patellar-trochlear compartment (OR, 1.70; P = .04) but not in the medial compartment (OR, 1.33; P = .23). There was an increased OR of Outerbridge grade 3 and 4 articular cartilage injury in patients from both groups having a prior medial meniscectomy on the medial femoral condyle (OR, 1.44; P < .01) and on the medial tibial plateau (OR, 1.63; P < .01). There was an increased OR of Outerbridge grade 3 and 4 articular cartilage injury in patients from both groups having a prior lateral meniscectomy on the lateral femoral condyle (OR, 1.65; P < .01) and on the lateral tibial plateau (OR, 1.56; P < .01). CONCLUSION: Meniscal tears are a common finding in both primary and revision ACL reconstruction. These results show a decreased OR of new untreated lateral meniscal tears in revision compared with primary ACL reconstruction. A previous medial or lateral meniscectomy increases the OR of articular cartilage damage in the medial or lateral compartments, respectively. Even when controlling for meniscus status, there is an increased OR in revision compared with primary ACL reconstruction of significant lateral compartment and patellar-trochlear chondral damage but not medial compartment chondral damage. PMID- 21646435 TI - Impact of intestinal glucuronidation on the pharmacokinetics of raloxifene. AB - Raloxifene is extensively glucuronidated in humans, effectively reducing its oral bioavailability (2%). It was also reported to be glucuronidated in preclinical animals, but its effects on the oral bioavailability have not been fully elucidated. In the present study, raloxifene and its glucuronides in the portal and systemic blood were monitored in Gunn rats deficient in UDP glucuronosyltransferase (UGT) 1A, Eisai hyperbilirubinemic rats (EHBRs), which hereditarily lack multidrug resistance-associated protein (MRP) 2, and wild-type rats after oral administration. The in vitro-in vivo correlation (IVIVC) of four UGT substrates (raloxifene, biochanin A, gemfibrozil, and mycophenolic acid) in rats was also evaluated. In Gunn rats, the product of fraction absorbed and intestinal availability and hepatic availability of raloxifene were 0.63 and 0.43, respectively; these values were twice those observed in wild-type Wistar rats, indicating that raloxifene was glucuronidated in both the liver and intestine. The ratio of glucuronides to unchanged drug in systemic blood was substantially higher in EHBRs (129-fold) than in the wild-type Sprague-Dawley rats (10-fold), suggesting the excretion of raloxifene glucuronides caused by MRP2. The IVIVC of the other UGT substrates in rats displayed a good relationship, but the oral clearance values of raloxifene and biochanin A, which were extensively glucuronidated by rat intestinal microsomes, were higher than the predicted clearances using rat liver microsomes, suggesting that intestinal metabolism may be a great contributor to the first-pass effect. Therefore, evaluation of intestinal and hepatic glucuronidation for new chemical entities is important to improve their pharmacokinetic profiles. PMID- 21646433 TI - Microbial ecology of the dark ocean above, at, and below the seafloor. AB - The majority of life on Earth--notably, microbial life--occurs in places that do not receive sunlight, with the habitats of the oceans being the largest of these reservoirs. Sunlight penetrates only a few tens to hundreds of meters into the ocean, resulting in large-scale microbial ecosystems that function in the dark. Our knowledge of microbial processes in the dark ocean-the aphotic pelagic ocean, sediments, oceanic crust, hydrothermal vents, etc.-has increased substantially in recent decades. Studies that try to decipher the activity of microorganisms in the dark ocean, where we cannot easily observe them, are yielding paradigm shifting discoveries that are fundamentally changing our understanding of the role of the dark ocean in the global Earth system and its biogeochemical cycles. New generations of researchers and experimental tools have emerged, in the last decade in particular, owing to dedicated research programs to explore the dark ocean biosphere. This review focuses on our current understanding of microbiology in the dark ocean, outlining salient features of various habitats and discussing known and still unexplored types of microbial metabolism and their consequences in global biogeochemical cycling. We also focus on patterns of microbial diversity in the dark ocean and on processes and communities that are characteristic of the different habitats. PMID- 21646436 TI - Characterization of the inhibitory effects of N-butylpyridinium chloride and structurally related ionic liquids on organic cation transporters 1/2 and human toxic extrusion transporters 1/2-k in vitro and in vivo. AB - Ionic liquids (ILs) are a class of salts that are expected to be used as a new source of solvents and for many other applications. Our previous studies revealed that selected ILs, structurally related organic cations, are eliminated exclusively in urine as the parent compound, partially mediated by renal transporters. This study investigated the inhibitory effects of N-butylpyridinium chloride (NBuPy-Cl) and structurally related ILs on organic cation transporters (OCTs) and multidrug and toxic extrusion transporters (MATEs) in vitro and in vivo. After Chinese hamster ovary cells expressing rat (r) OCT1, rOCT2, human (h) OCT2, hMATE1, or hMATE2-K were constructed, the ability of NBuPy-Cl, 1-methyl-3 butylimidazolium chloride (Bmim-Cl), N-butyl-N-methylpyrrolidinium chloride (BmPy Cl), and alkyl substituted pyridinium ILs to inhibit these transporters was determined in vitro. NBuPy-Cl (0, 0.5, or 2 mg/kg per hour) was also infused into rats to assess its effect on the pharmacokinetics of metformin, a substrate of OCTs and MATEs. NBuPy-Cl, Bmim-Cl, and BmPy-Cl displayed strong inhibitory effects on these transporters (IC(50) = 0.2-8.5 MUM). In addition, the inhibitory effects of alkyl-substituted pyridinium ILs on OCTs increased dramatically as the length of the alkyl chain increased. The IC(50) values were 0.1, 3.8, 14, and 671 MUM (hexyl-, butyl-, and ethyl-pyridinium and pyridinium chloride) for rOCT2 mediated metformin transport. Similar structurally related inhibitory kinetics were also observed for rOCT1 and hOCT2. The in vivo coadministration study revealed that NBuPy-Cl reduced the renal clearance of metformin in rats. These results demonstrate that ILs compete with other substrates of OCTs and MATEs and could alter the in vivo pharmacokinetics of such substrates. PMID- 21646432 TI - Genetic control of biosynthesis and transport of riboflavin and flavin nucleotides and construction of robust biotechnological producers. AB - Riboflavin [7,8-dimethyl-10-(1'-d-ribityl)isoalloxazine, vitamin B2] is an obligatory component of human and animal diets, as it serves as the precursor of flavin coenzymes, flavin mononucleotide, and flavin adenine dinucleotide, which are involved in oxidative metabolism and other processes. Commercially produced riboflavin is used in agriculture, medicine, and the food industry. Riboflavin synthesis starts from GTP and ribulose-5-phosphate and proceeds through pyrimidine and pteridine intermediates. Flavin nucleotides are synthesized in two consecutive reactions from riboflavin. Some microorganisms and all animal cells are capable of riboflavin uptake, whereas many microorganisms have distinct systems for riboflavin excretion to the medium. Regulation of riboflavin synthesis in bacteria occurs by repression at the transcriptional level by flavin mononucleotide, which binds to nascent noncoding mRNA and blocks further transcription (named the riboswitch). In flavinogenic molds, riboflavin overproduction starts at the stationary phase and is accompanied by derepression of enzymes involved in riboflavin synthesis, sporulation, and mycelial lysis. In flavinogenic yeasts, transcriptional repression of riboflavin synthesis is exerted by iron ions and not by flavins. The putative transcription factor encoded by SEF1 is somehow involved in this regulation. Most commercial riboflavin is currently produced or was produced earlier by microbial synthesis using special selected strains of Bacillus subtilis, Ashbya gossypii, and Candida famata. Whereas earlier RF overproducers were isolated by classical selection, current producers of riboflavin and flavin nucleotides have been developed using modern approaches of metabolic engineering that involve overexpression of structural and regulatory genes of the RF biosynthetic pathway as well as genes involved in the overproduction of the purine precursor of riboflavin, GTP. PMID- 21646437 TI - Metabolism and disposition of eltrombopag, an oral, nonpeptide thrombopoietin receptor agonist, in healthy human subjects. AB - The metabolism and disposition of eltrombopag, the first-in-class small molecule human thrombopoietin receptor agonist, were studied in six healthy men after a single oral administration of a solution dose of [(14)C]eltrombopag (75 mg, 100 MUCi). Eltrombopag was well tolerated. The drug was quickly absorbed and was the predominant circulating component in plasma (accounting for 63% of the total plasma radioactivity). A mono-oxygenation metabolite (M1) and acyl glucuronides (M2) of eltrombopag were minor circulating components. The predominant route of elimination of radioactivity was fecal (58.9%). Feces contained approximately 20% of dose as glutathione-related conjugates (M5, M6, and M7) and another 20% as unchanged eltrombopag. The glutathione conjugates were probably detoxification products of a p-imine methide intermediate formed by metabolism of M1, which arises through cytochrome P450-dependent processes. Low levels of covalently bound drug-related intermediates to plasma proteins, which could result from the reaction of the imine methide or acyl glucuronide conjugates with proteins, were detected. The bound material contributes to the longer plasma elimination half life of radioactivity. Renal elimination of conjugates of hydrazine cleavage metabolites (mostly as M3 and M4) accounted for 31% of the radiodose, with no unchanged eltrombopag detected in urine. PMID- 21646438 TI - Investigations of hydrazine cleavage of eltrombopag in humans. AB - After oral administration to humans, eltrombopag undergoes extensive cleavage of its hydrazine linkage to metabolites, which are exclusively eliminated in urine. In vitro, the cleavage pathway was not detected in systems using cytochrome P450 enzymes, renal or hepatic microsomes, or hepatocytes but was readily evident after anaerobic incubation with rodent cecal contents or human fecal homogenate. Antibiotic treatment in vitro and in vivo inhibited eltrombopag cleavage, further indicating that cleavage is via gut microbes. Antibiotic treatment did not alter the systemic exposure of eltrombopag in mice. Oral and intravenous pharmacokinetic characterization in the mice with one of the cleavage products indicated that it was readily absorbed, conjugated, and eliminated in urine, consistent with its fate after oral administration of eltrombopag. Variation in this microbial pathway, for example by antibiotic cotherapy, is unlikely to be clinically significant. PMID- 21646439 TI - From single compounds to herbal extract: a strategy to systematically characterize the metabolites of licorice in rats. AB - Because of the complicated chemical composition of traditional Chinese medicines (TCMs), their metabolic study has been a great challenge, especially when they are used in the traditional way, i.e., clinical oral dose of water decoction. Poor understanding of metabolic pathways and too low metabolite concentrations to be detected in biosamples are the major hurdles. In the present work, a three step strategy was proposed to systematically characterize in vivo metabolites of TCMs at a normal clinical dosage. Licorice, one of the most popular TCMs, was studied as a model. First, 10 representative compounds of licorice were administered to rats separately. A total of 68 metabolites were characterized by high-performance liquid chromatography coupled with diode-array detection and electrospray ionization tandem mass spectrometry and liquid chromatography (LC)/quadrupole time-of-flight-mass spectrometry (MS) analyses, together with enzyme hydrolysis. Among these, 13 compounds were confirmed by comparison with reference standards, including the 10 administered licorice compounds. Second, a high dose (equivalent to 20-fold clinical dosage) of licorice water extract was administered, and 22 more metabolites were characterized. Finally, these metabolites (including constituents of licorice) were determined by a highly sensitive and selective LC/selected reaction monitoring-MS method when the licorice water decoction was orally administered to rats at a clinical dosage (0.9 g crude drug/kg). A total of 42 metabolites in plasma and 62 metabolites in urine were detected. This is the first attempt to fully profile the in vivo metabolites of licorice at a normal clinical dosage. PMID- 21646440 TI - Influence of Panax ginseng on cytochrome P450 (CYP)3A and P-glycoprotein (P-gp) activity in healthy participants. AB - A number of herbal preparations have been shown to interact with prescription medications secondary to modulation of cytochrome P450 (CYP) and/or P glycoprotein (P-gp). The purpose of this study was to determine the influence of Panax ginseng on CYP3A and P-gp function using the probe substrates midazolam and fexofenadine, respectively. Twelve healthy participants (8 men) completed this open-label, single-sequence pharmacokinetic study. Healthy volunteers received single oral doses of midazolam 8 mg and fexofenadine 120 mg, before and after 28 days of P ginseng 500 mg twice daily. Midazolam and fexofenadine pharmacokinetic parameter values were calculated and compared before and after P ginseng administration. Geometric mean ratios (postginseng/preginseng) for midazolam area under the concentration-time curve from zero to infinity (AUC(0-infinity)), half life (t(1/2)), and maximum concentration (C(max)) were significantly reduced at 0.66 (0.55-0.78), 0.71 (0.53-0.90), and 0.74 (0.56-0.93), respectively. Conversely, fexofenadine pharmacokinetics were unaltered by P ginseng administration. Based on these results, P ginseng appeared to induce CYP3A activity in the liver and possibly the gastrointestinal tract. Patients taking P ginseng in combination with CYP3A substrates with narrow therapeutic ranges should be monitored closely for adequate therapeutic response to the substrate medication. PMID- 21646441 TI - Modeling longitudinal daily seizure frequency data from pregabalin add-on treatment. AB - The purpose of this study was to describe longitudinal daily seizure count data with respect to the effects of time and pregabalin add-on therapy. Models were developed in a stepwise manner: base model, time effect model, and time and drug effect (final) model, using a negative binomial distribution with Markovian features. Mean daily seizure count (lambda) was estimated to be 0.385 (relative standard error [RSE] 3.09%) and was further increased depending on the seizure count on the previous day. An overdispersion parameter (OVDP), representing extra Poisson variation, was estimated to be 0.330 (RSE 11.7%). Interindividual variances on lambda and OVDP were 84.7% and 210%, respectively. Over time, lambda tended to increase exponentially with a rate constant of 0.272 year-1 (RSE 26.8%). A mixture model was applied to classify responders/nonresponders to pregabalin treatment. Within the responders, lambda decreased exponentially with respect to dose with a constant of 0.00108 mg-1 (RSE 11.9%). The estimated responder rate was 66% (RSE 27.6%). Simulation-based diagnostics showed the model reasonably reproduced the characteristics of observed data. Highly variable daily seizure frequency was successfully characterized incorporating baseline characteristics, time effect, and the effect of pregabalin with classification of responders/nonresponders, all of which are necessary to adequately assess the efficacy of antiepileptic drugs. PMID- 21646442 TI - Assessment of the relative potency of fentanyl buccal tablet to intravenous morphine in healthy volunteers using a thermally induced hyperalgesia pain model. AB - This exploratory randomized, double-blind, placebo-controlled, 5-treatment, 5 period crossover study was conducted using a thermally induced hyperalgesia pain model in 51 healthy volunteers (33 evaluable) to characterize the relative potency of fentanyl buccal tablet (FBT) versus intravenous morphine. Relative potency was assessed using the sum of pain intensity differences over 60 minutes after the application of a 43 degrees C, 46 degrees C, and 49 degrees C painful stimulus following thermally induced hyperalgesia. Relative potency was also assessed by pupil diameter and responses to subjective questionnaires. The relative potency of FBT was 46.2 times that of intravenous morphine (95% confidence interval [CI], 17.6-575.3) based on the 49 degrees C stimulus. The relative potency of FBT based on opiate-induced miosis was 44.6 (95% CI, 29.7 77.0) at 60 minutes. These results are an initial relative potency assessment and should not be considered guidance for dose-equivalent switching between agents in clinical practice. PMID- 21646443 TI - Hereditary cerebellar degenerative disease (cerebellar cortical abiotrophy) in rabbits. AB - A pair of rabbits gave birth to a set of littermates (F1) with symptoms of early onset ataxia. Microscopic examination revealed cerebellar degenerative disease in 5 of 6 littermates. Light microscopy was used to compare the thickness of each cerebellar layer in affected animals in contrast to a normal control. Affected animals showed narrowing of the molecular layer of the vermis, reduced density of Purkinje cell dendrites and irregular thickness in their branchlets, and reduced density of granular cells and scattered pyknotic cells in the granular layer. Pyknotic cells were apoptotic granular cells, confirmed by positive staining using the TUNEL method. Electron microscopy confirmed the thinning of the molecular layer seen by light microscopy and also showed a reduced number of parallel fibers, which indicate granular cells axons, and a reduced number of synaptic junctions between Purkinje and granular cells. Purkinje cells had electron-dense, irregularly shaped cytoplasm with irregularly shaped nuclei, and some of these cells had a central chromatolysis-like region. These findings support a diagnosis of cerebellar cortical abiotrophy, a hereditary condition that causes nerve function impairment leading to early-onset progressive degeneration of the cerebellar cortex. PMID- 21646444 TI - Motor practice promotes increased activity in brain regions structurally disconnected after subcortical stroke. AB - BACKGROUND: Motor practice is an important component of neurorehabilitation. Imaging studies in healthy individuals show that dynamic brain activation changes with practice. Defining patterns of functional brain plasticity associated with motor practice following stroke could guide rehabilitation. OBJECTIVE: The authors aimed to test whether practice-related changes in brain activity differ after stroke and to explore spatial relationships between activity changes and patterns of structural degeneration. METHODS: They studied 10 patients at least 6 months after left-hemisphere subcortical strokes and 18 healthy controls. Diffusion-weighted magnetic resonance imaging (MRI) was acquired at baseline, and functional MRI (fMRI) was acquired during performance of a visuomotor tracking task before and after a 15-day period of practice of the same task. RESULTS: Smaller short-term practice effects at baseline correlated with lower fractional anisotropy in the posterior limbs of the internal capsule (PLIC) bilaterally in patients (t > 3; cluster P < .05). After 15 days of motor practice a Group * Time interaction (z > 2.3; cluster P < .05) was found in the basal ganglia, thalamus, inferior frontal gyrus, superior temporal gyrus, and insula. In these regions, healthy controls showed decreases and patients showed increases in activity with practice. Some regions of interest had a loss of white matter connectivity at baseline. CONCLUSIONS: Performance gains with motor practice can be associated with increased activity in regions that have been either directly or indirectly impaired by loss of connectivity. These results suggest that neurorehabilitation interventions may be associated with compensatory adaptation of intact brain regions as well as enhanced activity in regions with impaired structural connectivity. PMID- 21646445 TI - Greater sparing of visual search abilities in children after congenital rather than acquired focal brain damage. AB - BACKGROUND: Visual search refers to the capacity of an individual to find a target among simultaneously presented distracters and is based on visual abilities such as a fast visual processing and an accurate control of ballistic eye movements (saccades) that guide the fovea to the target location. OBJECTIVE: In adults, visual field defects caused by brain damage are often associated with visual search disorders; in children, little is known about the effects of early brain lesions on visual search abilities. METHODS: To test the presence of visual search defects and to investigate the role of cortical plasticity after early brain lesions, 29 children with congenital or acquired cerebral lesions, with and without visual field defects, underwent a visual search test battery. RESULTS: The children with acquired lesions and visual field defects had longer reaction times (RTs) in the contralesional visual field compared with the ipsilesional, whereas those with congenital lesions and visual field defects did not have differences in RTs between the contralateral and ipsilateral visual fields and had a visual search pattern similar to children without a visual field defect. CONCLUSIONS: These findings support the hypothesis of more effective mechanisms of functional compensation and reorganization of the visual system in children with very early brain lesions, as opposed to those with later damage. PMID- 21646446 TI - Serine proteases degrade airway mucins in cystic fibrosis. AB - Airway mucins are the major molecular constituents of mucus. Mucus forms the first barrier to invading organisms in the airways and is an important defense mechanism of the lung. We confirm that mucin concentrations are significantly decreased in airway secretions of subjects with cystic fibrosis (CF) who have chronic Pseudomonas aeruginosa infection. In sputum from CF subjects without a history of P. aeruginosa, we found no significant difference in the mucin concentration compared to mucus from normal controls. We demonstrate that mucins can be degraded by synthetic human neutrophil elastase (HNE) and P. aeruginosa elastase B (pseudolysin) and that degradation was inhibited by serine proteases inhibitors (diisopropyl fluorophosphates [DFP], phenylmethylsulfonyl fluoride [PMSF], and 1-chloro-3-tosylamido-7-amino-2-heptanone HCl [TLCK]). The mucin concentration in airway secretions from CF subjects is similar to that for normal subjects until there is infection by P. aeruginosa, and after that, the mucin concentration decreases dramatically. This is most likely due to degradation by serine proteases. The loss of this mucin barrier may contribute to chronic airway infection in the CF airway. PMID- 21646447 TI - Activation of adenosine A2A receptors inhibits neutrophil transuroepithelial migration. AB - Adenosine has been identified as a significant inhibitor of inflammation by acting on adenosine A(2A) receptors. In this study, we examined the role of adenosine and A(2A) receptors in the transmigration of human neutrophils across an in vitro model of the transitional bladder urothelium. Human uroepithelial cells (UROtsa) were grown on transwell inserts; uropathogenic Escherichia coli (UPEC) and neutrophils were added to the transwell system; and the number of migrating neutrophils was evaluated. Reverse transcription-PCR (RT-PCR), immunohistochemistry, and flow cytometry were used to investigate the expression of adenosine receptors, the epithelial adhesion molecule ICAM-1, and the neutrophil integrin CD11b. Levels of proinflammatory interleukin-8 (IL-8) and phosphorylated IkappaBalpha were measured by enzyme-linked immunosorbent assays (ELISA) and Luminex assays, respectively. The neutrophils expressed all four adenosine receptor subtypes (A(1), A(2A), A(2B), and A(3) receptors), but A(3) receptors were not expressed by UROtsa cells. UPEC stimulated neutrophil transuroepithelial migration, which was significantly decreased in response to the specific A(2A) receptor agonist CGS 21680. The inhibitory effect of CGS 21680 on neutrophil migration was reversed by the A(2A) receptor antagonist SCH 58261. The production of chemotactic IL-8 and the expression of the adhesion molecule ICAM-1 or CD11b were not significantly affected by CGS 21680. However, a significant decrease in the level of phosporylated IkappaBalpha was revealed in response to CGS 21680. In conclusion, UPEC infection in vitro evoked neutrophil migration through a multilayered human uroepithelium. The UPEC-evoked neutrophil transmigration decreased in response to A(2A) receptor activation, possibly through inhibition of NF-kappaB signaling pathways. PMID- 21646448 TI - Relative efficacy of uptake and presentation of Mycobacterium bovis BCG antigens by type I mouse lung epithelial cells and peritoneal macrophages. AB - Flow cytometric studies indicated that both peritoneal macrophages (PMs) and primary lung epithelial (PLE) cells isolated from mouse lungs could take up fluorescence-tagged Mycobacterium bovis BCG. BCG uptake in both cases was significantly inhibited by cytochalasin D, indicating active internalization of BCG by these cells. Confocal microscopy data further confirmed that BCG was internalized by PLE cells. BCG sonicate antigen (sBCG) had marked toxicity toward PMs but was relatively nontoxic to PLE cells. Accordingly, BCG sonicate antigen induced a significantly higher apoptotic and necrotic response in PMs compared to that in PLE cells. Both PMs and PLE cells exposed to BCG antigens and fixed thereafter could efficiently present antigens to purified BCG-sensitized T helper cells, as assessed by the release of interleukin-2 (IL-2) and gamma interferon (IFN-gamma). If, however, PLE cells were fixed before exposure to BCG, antigen presentation was abrogated, indicating that the PLE cells may in some way process the BCG antigen. A comparison of efficacies of BCG-pulsed PLE cells and PMs to present antigen at various antigen-presenting cell (APC)/T cell ratios indicated that PMs had only marginally greater APC function than that of PLE cells. Staining with specific monoclonal antibodies indicated that the cultured PLE cells used for antigen presentation essentially comprised type I epithelial cells. Our results suggest that type I lung epithelial cells may present BCG antigens to sensitized T helper cells and that their performance as APCs is comparable with that of PMs. PMID- 21646449 TI - Activation of the contact system at the surface of Fusobacterium necrophorum represents a possible virulence mechanism in Lemierre's syndrome. AB - Fusobacterium necrophorum causes Lemierre's syndrome, a serious disease with septic thrombophlebitis of the internal jugular vein, pulmonary involvement, and systemic inflammation. The contact system is a link between inflammation and coagulation, and contact activation by the bacteria could therefore contribute to the abnormal coagulation and inflammation seen in patients with Lemierre's syndrome. In this study, F. necrophorum was found to bind radiolabeled high molecular-weight kininogen (HK), a central component of the contact system. Binding was inhibited by the addition of unlabeled HK and domain D5 of HK but not other components of the contact system, indicating a specific interaction mediated through the D5 region. Binding of HK was significantly reduced after pretreatment of the bacteria with trypsin, suggesting that surface proteins are involved in HK binding. Incubation of the bacteria with human plasma resulted in an HK breakdown pattern suggestive of bradykinin release, and bradykinin was also detected in the supernatant. In addition, we show that factor XI (FXI), another component of the contact system, binds to F. necrophorum and that the bound FXI reconstitutes the activated partial thromboplastin time of FXI-deficient plasma. Thrombin activity was detected at the surface of the bacteria following incubation with plasma, indicating that the intrinsic pathway of coagulation is activated at the surface. This activity was completely blocked by inhibitors of the contact system. The combined results show that the contact system is activated at the surface of F. necrophorum, suggesting a pathogenic role for this system in Lemierre's syndrome. PMID- 21646450 TI - Expression of a Clostridium perfringens type IV pilin by Neisseria gonorrhoeae mediates adherence to muscle cells. AB - Clostridium perfringens is an anaerobic, Gram-positive bacterium that causes a range of diseases in humans, including lethal gas gangrene. We have recently shown that strains of C. perfringens move across the surface of agar plates by a unique type IV pilus (TFP)-mediated social motility that had not been previously described. Based on sequence homology to pilins in Gram-negative bacteria, C. perfringens appears to have two pilin subunits, PilA1 and PilA2. Structural prediction analysis indicated PilA1 is similar to the pseudopilin found in Klebsiella oxytoca, while PilA2 is more similar to true pilins found in the Gram negative pathogens Pseudomonas aeruginosa and Neisseria gonorrhoeae. Strains of N. gonorrhoeae that were genetically deficient in the native pilin, PilE, but supplemented with inducible expression of PilA1 and PilA2 of C. perfringens were constructed. Genetic competence, wild-type twitching motility, and attachment to human urogenital epithelial cells were not restored by expression of either pilin. However, attachment to mouse and rat myoblast (muscle) cell lines was observed with the N. gonorrhoeae strain expressing PilA2. Significantly, wild type C. perfringens cells adhered to mouse myoblasts under anaerobic conditions, and adherence was 10-fold lower in a pilT mutant that lacked functional TFP. These findings implicate C. perfringens TFP in the ability of C. perfringens to adhere to and move along muscle fibers in vivo, which may provide a therapeutic approach to limiting this rapidly spreading and highly lethal infection. PMID- 21646451 TI - Passive immunization with a polyclonal antiserum to the hemoglobin receptor of Haemophilus ducreyi confers protection against a homologous challenge in the experimental swine model of chancroid. AB - Haemophilus ducreyi, the etiologic agent of chancroid, has an obligate requirement for heme. Heme is acquired by H. ducreyi from its human host via TonB dependent transporters expressed at its bacterial surface. Of 3 TonB-dependent transporters encoded in the genome of H. ducreyi, only the hemoglobin receptor, HgbA, is required to establish infection during the early stages of the experimental human model of chancroid. Active immunization with a native preparation of HgbA (nHgbA) confers complete protection in the experimental swine model of chancroid, using either Freund's or monophosphoryl lipid A as adjuvants. To determine if transfer of anti-nHgbA serum is sufficient to confer protection, a passive immunization experiment using pooled nHgbA antiserum was conducted in the experimental swine model of chancroid. Pigs receiving this pooled nHgbA antiserum were protected from a homologous, but not a heterologous, challenge. Passively transferred polyclonal antibodies elicited to nHgbA bound the surface of H. ducreyi and partially blocked hemoglobin binding by nHgbA, but were not bactericidal. Taken together, these data suggest that the humoral immune response to the HgbA vaccine is protective against an H. ducreyi infection, possibly by preventing acquisition of the essential nutrient heme. PMID- 21646452 TI - Structure/function analysis of Neisseria meningitidis PilW, a conserved protein that plays multiple roles in type IV pilus biology. AB - Type IV pili (Tfp) are widespread filamentous bacterial organelles that mediate multiple functions and play a key role in pathogenesis in several important human pathogens, including Neisseria meningitidis. Tfp biology remains poorly understood at a molecular level because the roles of the numerous proteins that are involved remain mostly obscure. Guided by the high-resolution crystal structure we recently reported for N. meningitidis PilW, a widely conserved protein essential for Tfp biogenesis, we have performed a structure/function analysis by targeting a series of key residues through site-directed mutagenesis and analyzing the corresponding variants using an array of phenotypic assays. Here we show that PilW's involvement in the functionality of Tfp can be genetically uncoupled from its concurrent role in the assembly/stabilization of the secretin channels through which Tfp emerge on the bacterial surface. These findings suggest that PilW is a multifunctional protein. PMID- 21646453 TI - The Toll-like receptor 4 agonist monophosphoryl lipid a augments innate host resistance to systemic bacterial infection. AB - Monophosphoryl lipid A (MPLA) is a Toll-like receptor 4 (TLR4) agonist that is currently used as a vaccine adjuvant in humans. In this study, we evaluated the effect of MPLA treatment on the innate immune response to systemic bacterial infections in mice. Mice treated with MPLA after burn injury showed improved survival and less local and systemic dissemination of bacteria in a model of Pseudomonas aeruginosa burn wound infection. Prophylactic treatment with MPLA significantly enhanced bacterial clearance at the site of infection and reduced systemic dissemination of bacteria despite causing attenuation of proinflammatory cytokine production during acute intra-abdominal infection caused by cecal ligation and puncture. Administration of MPLA at 1 h after CLP also improved bacterial clearance but did not alter cytokine production. MPLA treatment increased the numbers of granulocytes, double-positive myeloid cells, and macrophages at sites of infection and increased the percentage and total numbers of myeloid cells mediating phagocytosis of bacteria. Depletion of Ly6G(+) neutrophils, but not macrophages, eliminated the ability of MPLA treatment to improve bacterial clearance. The immunomodulatory effects of MPLA were absent in TLR4-deficient mice. In conclusion, these studies show that MPLA treatment significantly augments the innate immune response to bacterial infection by enhancing bacterial clearance despite the attenuation of proinflammatory cytokine production. The enhanced bacterial clearance is mediated, in part, by increased numbers of myeloid cells with effective phagocytic functions at sites of infection and is TLR4 dependent. PMID- 21646454 TI - Direct evaluation of Pseudomonas aeruginosa biofilm mediators in a chronic infection model. AB - Biofilms contribute to Pseudomonas aeruginosa persistence in a variety of diseases, including cystic fibrosis, burn wounds, and chronic suppurative otitis media. However, few studies have directly addressed P. aeruginosa biofilms in vivo. We used a chinchilla model of otitis media, which has previously been used to study persistent Streptococcus pneumoniae and Haemophilus influenzae infections, to show that structures formed in vivo are biofilms of bacterial and host origin within a matrix that includes Psl, a P. aeruginosa biofilm polysaccharide. We evaluated three biofilm and/or virulence mediators of P. aeruginosa known to affect biofilm formation in vitro and pathogenesis in vivo- bis-(3',5')-cyclic dimeric GMP (c-di-GMP), flagella, and quorum sensing--in a chinchilla model. We show that c-di-GMP overproduction has a positive impact on bacterial persistence, while quorum sensing increases virulence. We found no difference in persistence attributed to flagella. We conclude from these studies that a chinchilla otitis media model provides a means to evaluate pathogenic mediators of P. aeruginosa and that in vitro phenotypes should be examined in multiple infection systems to fully understand their role in disease. PMID- 21646456 TI - Bacteriophage-encoding cytolethal distending toxin type V gene induced from nonclinical Escherichia coli isolates. AB - Cytolethal distending toxin (Cdt) is produced by a variety of pathogenic bacteria, including pathogenic serotypes of Shiga toxin-producing Escherichia coli (STEC). The Cdt family comprises five variants (Cdt-I to Cdt-V) encoded by three genes located within the chromosome or plasmids or, in the case of Cdt-I, within bacteriophages. In this study, we evaluated the occurrence of the cdt gene in a collection of 140 environmental STEC isolates. cdt was detected in 12.1% of strains, of which five strains carried inducible bacteriophages containing the Cdt-V variant. Two Cdt-V phages of the Siphoviridae morphology lysogenized Shigella sonnei, generating two lysogens: a single Cdt phage lysogen and a double lysogen, containing a Cdt phage and an Stx phage, both from the wild-type strain. The rates of induction of Cdt phages were evaluated by quantitative PCR, and spontaneous induction of Cdt-V phage was observed, whereas induction of Stx phage in the double lysogen was mitomycin C dependent. The Cdt distending effect was observed in HeLa cells inoculated with the supernatant of the Cdt-V phage lysogen. A ClaI fragment containing the cdt-V gene of one phage was cloned, and sequencing confirmed the presence of Cdt-V, as well as a fragment downstream from the cdt homolog to gpA, encoding a replication protein of bacteriophage P2. Evaluation of Cdt-V phages in nonclinical water samples showed densities of 10(2) to 10(9) gene copies in 100 ml, suggesting the high prevalence of Cdt phages in nonclinical environments. PMID- 21646458 TI - Steps forward no guarantee that health targets will be met, council says. PMID- 21646459 TI - Legal ambiguities surround authority to make end-of-life decisions. PMID- 21646455 TI - Bacterial virulence in the moonlight: multitasking bacterial moonlighting proteins are virulence determinants in infectious disease. AB - Men may not be able to multitask, but it is emerging that proteins can. This capacity of proteins to exhibit more than one function is termed protein moonlighting, and, surprisingly, many highly conserved proteins involved in metabolic regulation or the cell stress response have a range of additional biological actions which are involved in bacterial virulence. This review highlights the multiple roles exhibited by a range of bacterial proteins, such as glycolytic and other metabolic enzymes and molecular chaperones, and the role that such moonlighting activity plays in the virulence characteristics of a number of important human pathogens, including Staphylococcus aureus, Streptococcus pyogenes, Streptococcus pneumoniae, Helicobacter pylori, and Mycobacterium tuberculosis. PMID- 21646461 TI - Small step forward in global food labelling. PMID- 21646463 TI - Candida parapsilosis in the blood smear of an injection drug user. PMID- 21646462 TI - Prospective validation of the ABCD2 score for patients in the emergency department with transient ischemic attack. AB - BACKGROUND: The ABCD2 score (Age, Blood pressure, Clinical features, Duration of symptoms and Diabetes) is used to identify patients having a transient ischemic attack who are at high risk for imminent stroke. However, despite its widespread implementation, the ABCD2 score has not yet been prospectively validated. We assessed the accuracy of the ABCD2 score for predicting stroke at 7 (primary outcome) and 90 days. METHODS: This prospective cohort study enrolled adults from eight Canadian emergency departments who had received a diagnosis of transient ischemic attack. Physicians completed data forms with the ABCD2 score before disposition. The outcome criterion, stroke, was established by a treating neurologist or by an Adjudication Committee. We calculated the sensitivity and specificity for predicting stroke 7 and 90 days after visiting the emergency department using the original "high-risk" cutpoint of an ABCD2 score of more than 5, and the American Heart Association recommendation of a score of more than 2. RESULTS: We enrolled 2056 patients (mean age 68.0 yr, 1046 (50.9%) women) who had a rate of stroke of 1.8% at 7 days and 3.2% at 90 days. An ABCD2 score of more than 5 had a sensitivity of 31.6% (95% confidence interval [CI] 19.1-47.5) for stroke at 7 days and 29.2% (95% CI 19.6-41.2) for stroke at 90 days. An ABCD2 score of more than 2 resulted in sensitivity of 94.7% (95% CI 82.7-98.5) for stroke at 7 days with a specificity of 12.5% (95% CI 11.2-14.1). The accuracy of the ABCD2 score as calculated by either the enrolling physician (area under the curve 0.56; 95% CI 0.47-0.65) or the coordinating centre (area under the curve 0.65; 95% CI 0.57-0.73) was poor. INTERPRETATION: This multicentre prospective study involving patients in emergency departments with transient ischemic attack found the ABCD2 score to be inaccurate, at any cut-point, as a predictor of imminent stroke. Furthermore, the ABCD2 score of more than 2 that is recommended by the American Heart Association is nonspecific. PMID- 21646464 TI - Administrator urges penalties for doctors who don't use electronic medical records. PMID- 21646465 TI - Acute primary HIV infection. PMID- 21646466 TI - National organ registry and allocation system remains remote. PMID- 21646467 TI - Preventing stroke after transient ischemic attack. PMID- 21646468 TI - Effect of ON 01910.Na, an anticancer mitotic inhibitor, on cell-cycle progression correlates with RanGAP1 hyperphosphorylation. AB - The benzyl styryl sulfone, ON 01910.Na, is a novel anticancer agent that inhibits mitotic progression and induces apoptosis in most cancer cell lines. We examined the effect of ON 01910.Na on DNA damage-signaling molecules upstream of Cdc25C (Chk1, Chk2, and H2AX), as well as on Ran GTPase-activating protein 1 conjugated to small ubiquitin-related modifier 1 (RanGAP1.SUMO1), a mitosis coordinator. Prostate cancer, lymphoma, and leukemic cells were incubated with the drug for 4, 16, or 24 hours. Cell lysates were resolved on SDS-PAGE and analyzed by Western blot. Camptothecin and doxorubicin treatment caused activation/phosphorylation of DNA damage-responsive molecules by 4 hours, whereas ON 01910.Na did not do so. ON 01910.Na caused hyperphosphorylation of RanGAP1.SUMO1 within 4 hours that was sustained for more than 24 hours. Mild phosphorylation of Chk2 was observed only after 24-hour exposure, indicating that DNA damage response was not an initial effect of ON 01910.Na. MOLT-3 cells, synchronized by double-thymidine block, when released into a medium containing ON 01910.Na, accumulated mitotic cell number with a peak from 10 to 14 hours and remained near plateau for 20 hours, which corresponded with the time of RanGAP phosphorylation. ON 01910.Na had minimal effects on tubulin polymerization. These findings imply that ON 01910.Na neither induces DNA damage directly nor acts as a tubulin toxin. Its biological activity appears to rely on prolonged phosphorylation/hyperphosphorylation of RanGAP1.SUMO1. M-phase arrest and the consequent induction of apoptosis that follows could possibly be attributed to it. ON 01910.Na may act as an inhibitor of a RanGAP1.SUMO1 phosphatase or a stimulant of a new kinase. RanGAP1.SUMO1 appears to be a new target pathway for cancer chemotherapy. PMID- 21646469 TI - Dissecting the oncogenic and tumorigenic potential of differentiated human induced pluripotent stem cells and human embryonic stem cells. AB - Pluripotent stem cells, both human embryonic stem cells (hESC) and human-induced pluripotent stem cells (hiPSC), can give rise to multiple cell types and hence have tremendous potential for regenerative therapies. However, the tumorigenic potential of these cells remains a great concern, as reflected in the formation of teratomas by transplanted pluripotent cells. In clinical practice, most pluripotent cells will be differentiated into useful therapeutic cell types such as neuronal, cardiac, or endothelial cells prior to human transplantation, drastically reducing their tumorigenic potential. Our work investigated the extent to which these differentiated stem cell derivatives are truly devoid of oncogenic potential. In this study, we analyzed the gene expression patterns from three sets of hiPSC- and hESC-derivatives and the corresponding primary cells, and compared their transcriptomes with those of five different types of cancer. Our analysis revealed a significant gene expression overlap of the hiPSC- and hESC-derivatives with cancer, whereas the corresponding primary cells showed minimum overlap. Real-time quantitative PCR analysis of a set of cancer-related genes (selected on the basis of rigorous functional and pathway analyses) confirmed our results. Overall, our findings suggested that pluripotent stem cell derivatives may still bear oncogenic properties even after differentiation, and additional stringent functional assays to purify these cells should be done before they can be used for regenerative therapy. PMID- 21646470 TI - Notch signaling in CD66+ cells drives the progression of human cervical cancers. AB - Human epithelial tumor progression and metastasis involve cellular invasion, dissemination in the vasculature, and regrowth at metastatic sites. Notch signaling has been implicated in metastatic progression but its roles have yet to be fully understood. Here we report the important role of Notch signaling in maintaining cells expressing the carcinoembryonic antigen cell adhesion molecule CEACAM (CD66), a known mediator of metastasis. CD66 and Notch1 were studied in clinical specimens and explants of human cervical cancer, including specimens grown in a pathophysiologically relevant murine model. Gene expression profiling of CD66(+) cells from primary tumors showed enhanced features of Notch signaling, metastasis, and stemness. Significant differences were also seen in invasion, colony formation, and tumor forming efficiency between CD66(+) and CD66(-) cancer cells. Notably, CD66(+) cells showed a marked sensitivity to a Notch small molecule inhibitor. In support of studies in established cell lines, we documented the emergence of a tumorigenic CD66(+) cell subset within a metastatic lesion-derived cervical-cancer cell line. Similar to primary cancers, CD66 expression in the cell line was blocked by chemical and genetic inhibitors of ligand-dependent nuclear Notch signaling. Collectively, our work on the oncogenic properties of CD66(+) cells in epithelial cancers provides insights into the nature of tumor progression and offers a mechanistic rationale to inhibit the Notch signaling pathway as a generalized therapeutic strategy to treat metastatic cancers. PMID- 21646471 TI - Small-molecule anticancer compounds selectively target the hemopexin domain of matrix metalloproteinase-9. AB - Lack of target specificity by existing matrix metalloproteinase (MMP) inhibitors has hindered antimetastatic cancer drug discovery. Inhibitors that bind to noncatalytic sites of MMPs and disrupt protease signaling function have the potential to be more specific and selective. In this work, compounds that target the hemopexin (PEX) domain of MMP-9 were identified using an in silico docking approach and evaluated using biochemical and biological approaches. Two of the selected compounds interfere with MMP-9-mediated cancer cell migration and proliferation in cells expressing exogenous or endogenous MMP-9. Furthermore, these inhibitors do not modulate MMP-9 catalytic activity. The lead compound, N [4-(difluoromethoxy)phenyl]-2-[(4-oxo-6-propyl-1H-pyrimidin-2-yl)sulfanyl] acetamide, specifically binds to the PEX domain of MMP-9, but not other MMPs. This interaction between the compound and the PEX domain results in the abrogation of MMP-9 homodimerization and leads to blockage of a downstream signaling pathway required for MMP-9-mediated cell migration. In a tumor xenograft model, this pyrimidinone retarded MDA-MB-435 tumor growth and inhibited lung metastasis. Thus, we have shown for the first time that a novel small molecule interacts specifically with the PEX domain of MMP-9 and inhibits tumor growth and metastasis by reducing cell migration and proliferation. PMID- 21646472 TI - Overcoming hypoxia-induced apoptotic resistance through combinatorial inhibition of GSK-3beta and CDK1. AB - Tumor hypoxia is an inherent impediment to cancer treatment that is both clinically significant and problematic. In this study, we conducted a cell-based screen to identify small molecules that could reverse the apoptotic resistance of hypoxic cancer cells. Among the compounds, we identified were a structurally related group that sensitized hypoxic cancer cells to apoptosis by inhibiting the kinases GSK-3beta and cyclin-dependent kinase (CDK) 1. Combinatorial inhibition of these proteins in hypoxic cancer cells and tumors increased levels of c-Myc and decreased expression of c-IAP2 and the central hypoxia response regulator hypoxia-inducible factor (HIF) 1alpha. In mice, these compounds augmented the hypoxic tumor cell death induced by cytotoxic chemotherapy, blocking angiogenesis and tumor growth. Taken together, our findings suggest that combinatorial inhibition of GSK-3beta and CDK1 augment the apoptotic sensitivity of hypoxic tumors, and they offer preclinical validation of a novel and readily translatable strategy to improve cancer therapy. PMID- 21646473 TI - High phosphoantigen levels in bisphosphonate-treated human breast tumors promote Vgamma9Vdelta2 T-cell chemotaxis and cytotoxicity in vivo. AB - The nitrogen-containing bisphosphonate zoledronic acid (ZOL), a potent inhibitor of farnesyl pyrophosphate synthase, blocks the mevalonate pathway, leading to intracellular accumulation of isopentenyl pyrophosphate/triphosphoric acid I adenosin-5'-yl ester 3-(3-methylbut-3-enyl) ester (IPP/ApppI) mevalonate metabolites. IPP/ApppI accumulation in ZOL-treated cancer cells may be recognized by Vgamma9Vdelta2 T cells as tumor phosphoantigens in vitro. However, the significance of these findings in vivo remains largely unknown. In this study, we investigated the correlation between the anticancer activities of Vgamma9Vdelta2 T cells and the intracellular IPP/ApppI levels in ZOL-treated breast cancer cells in vitro and in vivo. We found marked differences in IPP/ApppI production among different human breast cancer cell lines post-ZOL treatment. Coculture with purified human Vgamma9Vdelta2 T cells led to IPP/ApppI-dependent near-complete killing of ZOL-treated breast cancer cells. In ZOL-treated mice bearing subcutaneous breast cancer xenografts, Vgamma9Vdelta2 T cells infiltrated and inhibited growth of tumors that produced high IPP/ApppI levels, but not those expressing low IPP/ApppI levels. Moreover, IPP/ApppI not only accumulated in cancer cells but it was also secreted, promoting Vgamma9Vdelta2 T-cell chemotaxis to the tumor. Without Vgamma9Vdelta2 T-cell expansion, ZOL did not inhibit tumor growth. These findings suggest that cancers-producing high IPP/ApppI levels after ZOL treatment are most likely to benefit from Vgamma9Vdelta2 T-cell-mediated immunotherapy. PMID- 21646474 TI - Pivotal role of innate and adaptive immunity in anthracycline chemotherapy of established tumors. AB - We show, in a series of established experimental breast adenocarcinomas and fibrosarcomas induced by carcinogen de novo in mice, that the therapeutic efficacy of doxorubicin treatment is dependent on CD8 T cells and IFN-gamma production. Doxorubicin treatment enhances tumor antigen-specific proliferation of CD8 T cells in tumor-draining lymph nodes and promotes tumor infiltration of activated, IFN-gamma-producing CD8 T cells. Optimal doxorubicin treatment outcome also requires both interleukin (IL)-1beta and IL-17 cytokines, as blockade of IL 1beta/IL-1R or IL-17A/IL-17Ralpha signaling abrogated the therapeutic effect. IL 23p19 had no observed role. The presence of gammadelta T cells, but not Jalpha18(+) natural killer T cells, at the time of doxorubicin treatment was also important. In tumor samples taken from breast cancer patients prior to treatment with anthracycline chemotherapy, a correlation between CD8alpha, CD8beta, and IFN gamma gene expression levels and clinical response was observed, supporting their role in the therapeutic efficacy of anthracyclines in humans. Overall, these data strongly support the pivotal contribution of both innate and adaptive immunity in treatment outcomes of anthracycline chemotherapy. PMID- 21646475 TI - 18F-fluorodeoxy-glucose positron emission tomography marks MYC-overexpressing human basal-like breast cancers. AB - In contrast to normal cells, cancer cells avidly take up glucose and metabolize it to lactate even when oxygen is abundant, a phenomenon referred to as the Warburg effect. This fundamental alteration in glucose metabolism in cancer cells enables their specific detection by positron emission tomography (PET) following i.v. injection of the glucose analogue (18)F-fluorodeoxy-glucose ((18)FDG). However, this useful imaging technique is limited by the fact that not all cancers avidly take up FDG. To identify molecular determinants of (18)FDG retention, we interrogated the transcriptomes of human-cancer cell lines and primary tumors for metabolic pathways associated with (18)FDG radiotracer uptake. From ninety-five metabolic pathways that were interrogated, the glycolysis, and several glycolysis-related pathways (pentose phosphate, carbon fixation, aminoacyl-tRNA biosynthesis, one-carbon-pool by folate) showed the greatest transcriptional enrichment. This "FDG signature" predicted FDG uptake in breast cancer cell lines and overlapped with established gene expression signatures for the "basal-like" breast cancer subtype and MYC-induced tumorigenesis in mice. Human breast cancers with nuclear MYC staining and high RNA expression of MYC target genes showed high (18)FDG-PET uptake (P < 0.005). Presence of the FDG signature was similarly associated with MYC gene copy gain, increased MYC transcript levels, and elevated expression of metabolic MYC target genes in a human breast cancer genomic dataset. Together, our findings link clinical observations of glucose uptake with a pathologic and molecular subtype of human breast cancer. Furthermore, they suggest related approaches to derive molecular determinants of radiotracer retention for other PET-imaging probes. PMID- 21646476 TI - Memory type 2 helper T cells induce long-lasting antitumor immunity by activating natural killer cells. AB - Functionally polarized helper T cells (Th cells) play crucial roles in the induction of tumor immunity. There is considerable knowledge about the contributions of IFN-producing Th1 cells that supports the role of cytotoxic cluster of differentiation (CD8) T cells and natural killer (NK) cells, but much less is known about how IL-4-producing Th2 cells contribute to tumor immunity. In this study, we investigated the cellular and molecular mechanisms employed by memory Th2 cells in sustaining tumor immunity by using a mouse model system wherein ovalbumin (OVA) is used as a specific tumor antigen. In this model, we found that OVA-specific memory Th2 cells exerted potent and long-lasting antitumor effects against NK-sensitive OVA-expressing tumor cells, wherein antitumor effects were mediated by NK cells. Specifically, NK cell cytotoxic activity and expression of perforin and granzyme B were dramatically enhanced by the activation of memory Th2 cells. Interleukin 4 (IL-4) produced by memory Th2 cells in vivo was critical for the antitumor effects of the NK cells, which IL-4 directly stimulated to induce their perforin- and granzyme-B-dependent cytotoxic activity. Our findings show that memory Th2 cells can induce potent antitumor immunity through IL-4-induced activation of NK cells, suggesting potential applications in cellular therapy for cancer patients. PMID- 21646477 TI - Ex vivo graft purging and expansion of autologous blood progenitor cell products from patients with multiple myeloma. AB - Autologous peripheral blood progenitor cell (PBPC) transplantation is the treatment of choice for selected myeloma patients. However, tumor cells contaminating the apheresis product are a potential source of relapse. Here we report a sequential purging strategy targeting mature and immature clonogenic myeloma cell populations in the autograft. Thawed PBPC products of myeloma patients were treated with rituximab to kill CD138(-)20(+) B cells (highly clonogenic immature cells), and bortezomib to target CD138(+) cells (normal and differentiated myeloma plasma cells), followed by coculture with allogeneic mesenchymal stem cells (MSC) from normal donors. After 7 days of coculture, nonadherent cells were removed and cultured in the absence of MSC for an additional 7 days. Then, efficacy of purging (removal of CD138(-)20(+) and CD138(+) cells) was assessed by flow cytometry and PCR. We used our ex vivo purging strategy to treat frozen aphereses from 16 patients. CD138(+) and CD138( )20(+)(19(+)) cells present in the initial products were depleted more than 3 and 4 logs, respectively based on 10(6) flow-acquisition events, and to levels below the limit of detection by PCR. In contrast, total nucleated cell (TNC), CD34(+) cell, and colony-forming cell numbers were increased by approximately 12 to 20, 8 , and 23-fold, respectively. Overall, ex vivo treatment of apheresis products with rituximab, bortezomib, and coculture with normal donor MSC depleted mature and immature myeloma cells from clinical aphereses while expanding the normal hematopoietic progenitor cell compartment. PMID- 21646478 TI - Sequential activation of Snail1 and N-Myc modulates sonic hedgehog-induced transformation of neural cells. AB - Activation of the Sonic hedgehog (Shh) pathway and increased expression of Gli1 play an important role in proliferation and transformation of granule cell progenitors (GCP) in the developing cerebellum. Medulloblastomas arising from cerebellar GCPs are frequently driven by Shh pathway-activating mutations; however, molecular mechanisms of Shh pathway dysregulation and transformation of neural progenitors remain poorly defined. We report that the transcription factor and oncogene Snail1 (Sna1) is directly induced by Shh pathway activity in GCPs, murine medulloblastomas, and human medulloblastoma cells. Enforced expression of Sna1 was sufficient to induce GCPs and medulloblastoma cell proliferation in the absence of Shh/Gli1 exposure. In addition, enforced expression of Sna1 increased transformation of medulloblastoma cells in vitro and in vivo. Analysis of potential Sna1 targets in neural cells revealed a novel Sna1 target, N-Myc, a transcription factor known to play a role in Shh-mediated GCP proliferation and medulloblastoma formation. We found that Sna1 directly induced transcription of N Myc in human medulloblastoma cells and that depletion of N-Myc ablated the Sna1 induced proliferation and transformation. Taken together, these results provide further insight into the mechanism of Shh-induced transformation of neural progenitor cells and suggest that induction of Sna1 may serve to amplify the oncogenic potential of Shh pathway activation through N-Myc induction. PMID- 21646479 TI - Sitamaquine overcomes ABC-mediated resistance to miltefosine and antimony in Leishmania. AB - Although oral miltefosine represented an important therapeutic advance in the treatment of leishmaniasis, the appearance of resistance remains a serious threat. LMDR1/LABCB4, a P-glycoprotein-like transporter included in the Leishmania ABC (ATP-binding cassette) family, was the first molecule shown to be involved in experimental miltefosine resistance. LMDR1 pumps drugs out of the parasite, thereby decreasing their intracellular accumulation. Sitamaquine, another promising oral drug for leishmaniasis, is currently in phase 2b clinical trials. The physicochemical features of this drug suggested to us that it could be considered for use as an LMDR1 inhibitor. Indeed, we report herein that nonleishmanicidal concentrations of sitamaquine reverse miltefosine resistance in a multidrug resistance Leishmania tropica line that overexpresses LMDR1. This reversal effect is due to modulation of the LMDR1-mediated efflux of miltefosine. In addition, sitamaquine is not a substrate of LMDR1, as this transporter does not affect sitamaquine accumulation or sensitivity in the parasite. Likewise, we show that ketoconazole, another oral leishmanicidal drug known to interact with ABC transporters, is also able to reverse LMDR1-mediated miltefosine resistance, although with a lower efficiency than sitamaquine. Molecular docking on a three dimensional homology model of LMDR1 showed different preferential binding sites for each substrate-inhibitor pair, thus explaining this different behavior. Finally, we show that sitamaquine is also able to modulate the antimony resistance mediated by MRPA/LABCC3, another ABC transporter involved in experimental and clinical antimony resistance in this parasite. Taken together, these data suggest that the combination of sitamaquine with miltefosine or antimony could avoid the appearance of resistance mediated by these membrane transporters in Leishmania. PMID- 21646480 TI - The base component of 3'-azido-2',3'-dideoxynucleosides influences resistance mutations selected in HIV-1 reverse transcriptase. AB - We recently reported that HIV-1 resistant to 3'-azido-3'-deoxythymidine (AZT) is not cross-resistant to 3'-azido-2',3'-dideoxypurines. This finding suggested that the nucleoside base is a major determinant of HIV-1 resistance to nucleoside analogs. To further explore this hypothesis, we conducted in vitro selection experiments by serial passage of HIV-1(LAI) in MT-2 cells in increasing concentrations of 3'-azido-2',3'-dideoxyguanosine (3'-azido-ddG), 3'-azido-2',3' dideoxycytidine (3'-azido-ddC), or 3'-azido-2',3'-dideoxyadenosine (3'-azido ddA). 3'-Azido-ddG selected for virus that was 5.3-fold resistant to 3'-azido-ddG compared to wild-type HIV-1(LAI) passaged in the absence of drug. Population sequencing of the entire reverse transcriptase (RT) gene identified L74V, F77L, and L214F mutations in the polymerase domain and K476N and V518I mutations in the RNase H domain. However, when introduced into HIV-1 by site-directed mutagenesis, these 5 mutations only conferred ~2.0-fold resistance. Single-genome sequencing analyses of the selected virus revealed a complex population of mutants that all contained L74V and L214F linked to other mutations, including ones not identified during population sequencing. Recombinant HIV-1 clones containing RT derived from single sequences exhibited 3.2- to 4.0-fold 3'-azido-ddG resistance. In contrast to 3'-azido-ddG, 3'-azido-ddC selected for the V75I mutation in HIV-1 RT that conferred 5.9-fold resistance, compared to the wild-type virus. Interestingly, we were unable to select HIV-1 that was resistant to 3'-azido-ddA, even at concentrations of 3'-azido-ddA that yielded high intracellular levels of 3'-azido ddA-5'-triphosphate. Taken together, these findings show that the nucleoside base is a major determinant of HIV-1 resistance mechanisms that can be exploited in the design of novel nucleoside RT inhibitors. PMID- 21646481 TI - Novel pentadecenyl tetrazole enhances susceptibility of methicillin-resistant Staphylococcus aureus biofilms to gentamicin. AB - One method that bacteria employ to reduce their susceptibility to antibiotics is the formation of biofilms. We developed a robust 6-well plate biofilm assay to evaluate early-stage discovery compounds against methicillin-resistant Staphylococcus aureus (MRSA). Tissue culture-treated 6-well plates were selected for this assay because they facilitate the adherence of MRSA and enable accurate determination of the number of CFU in each well. The MRSA biofilms formed in this assay exhibit increased tolerances to clinically used antibiotics. Using this biofilm assay, we identified a novel potentiator of gentamicin against MRSA biofilms. The combination of gentamicin and pentadecenyl tetrazole is superior to clinically used MRSA antibiotics against these MRSA biofilms. This novel combination also exhibits synergistic effects on MRSA planktonic cells. This plant-derived compound reveals promise for its effectiveness and warrants further lead optimization as an antibiotic and aminoglycoside potentiator. PMID- 21646482 TI - The pmrCAB operon mediates polymyxin resistance in Acinetobacter baumannii ATCC 17978 and clinical isolates through phosphoethanolamine modification of lipid A. AB - The emergence of multidrug resistance among Acinetobacter baumannii is leading to an increasing dependence on the use of polymyxins as last-hope antibiotics. Here, we utilized genetic and biochemical methods to define the involvement of the pmrCAB operon in polymyxin resistance in this organism. Sequence analysis of 16 polymyxin B-resistant strains, including 6 spontaneous mutants derived from strain ATCC 17978 and 10 clinical isolates from diverse sources, revealed that they had independent mutations in the pmrB gene, encoding a sensor kinase, or in the response regulator PmrA. Knockout of the pmrB gene in two mutants and two clinical isolates led to a decrease in the polymyxin B susceptibility of these strains, which could be restored with the cloned pmrAB genes from the mutants but not from the wild type. Reverse transcription-quantitative PCR (RT-qPCR) analysis also showed a correlation between the expression of pmrC and polymyxin B resistance. Characterization of lipid A species from the mutant strains, by thin layer chromatography and mass spectrometry, indicated that the addition of phosphoethanolamine to lipid A correlated with resistance. This addition is performed in Salmonella enterica serovar Typhimurium by the product of the pmrC gene, which is a homolog of the pmrC gene from Acinetobacter. Knockout of this gene in the mutant R2 [pmrB(T235I)] reversed resistance as well as phosphoethanolamine modification of lipid A. These results demonstrate that specific alterations in the sequence of the pmrCAB operon are responsible for resistance to polymyxins in A. baumannii. PMID- 21646483 TI - Low fitness cost of the multidrug resistance gene cfr. AB - The recently described rRNA methyltransferase Cfr that methylates the conserved 23S rRNA residue A2503, located in a functionally critical region of the ribosome, confers resistance to an array of ribosomal antibiotics, including linezolid. A number of reports of linezolid-resistant cfr-positive clinical strains indicate the possible rapid spread of this resistance mechanism. Since the rate of dissemination and the efficiency of maintenance of a resistance gene depend on the fitness cost associated with its acquisition, we investigated the fitness cost of cfr expression in a laboratory Staphylococcus aureus strain. We found that acquisition of the cfr gene does not produce any appreciable reduction in the cell growth rate. Only in a cogrowth competition experiment was some loss of fitness observed because Cfr-expressing cells slowly lose to the cfr-negative control strain. Interestingly, cells expressing wild-type and catalytically inactive Cfr had very similar growth characteristics, indicating that the slight fitness cost associated with cfr acquisition stems from expression of the Cfr polypeptide rather than from the modification of the conserved rRNA residue. In some clinical isolates, cfr is coexpressed with the erm gene, which encodes a methyltransferase targeting another 23S rRNA residue, A2058. Dimethylation of A2058 by Erm notably increases the fitness cost associated with the Cfr-mediated methylation of A2503. The generally low fitness cost of cfr acquisition observed in our experiments with the laboratory S. aureus strain offers a microbiological explanation for the apparent spread of the cfr gene among pathogens. PMID- 21646484 TI - Antiplasmodial properties of acyl-lysyl oligomers in culture and animal models of malaria. AB - Our previous analysis of antiplasmodial properties exhibited by dodecanoyl-based oligo-acyl-lysyls (OAKs) has outlined basic attributes implicated in potent inhibition of parasite growth and underlined the critical role of excess hydrophobicity in hemotoxicity. To dissociate hemolysis from antiplasmodial effect, we screened >50 OAKs for in vitro growth inhibition of Plasmodium falciparum strains, thus revealing the minimal requirements for antiplasmodial potency in terms of sequence and composition, as confirmed by efficacy studies in vivo. The most active sequence, dodecanoyllysyl-bis(aminooctanoyllysyl)-amide (C(12)K-2alpha(8)), inhibited parasite growth at submicromolar concentrations (50% inhibitory concentration [IC(50)], 0.3 +/- 0.1 MUM) and was devoid of hemolytic activity (<0.4% hemolysis at 150 MUM). Unlike the case of dodecanoyl based analogs, which equally affect ring and trophozoite stages of the parasite developmental cycle, the ability of various octanoyl-based OAKs to distinctively affect these stages (rings were 4- to 5-fold more sensitive) suggests a distinct antiplasmodial mechanism, nonmembranolytic to host red blood cells (RBCs). Upon intraperitoneal administration to mice, C(12)K-2alpha(8) demonstrated sustainable high concentrations in blood (e.g., 0.1 mM at 25 mg/kg of body weight). In Plasmodium vinckei-infected mice, C(12)K-2alpha(8) significantly affected parasite growth (50% effective dose [ED(50)], 22 mg/kg) but also caused mortality in 2/3 mice at high doses (50 mg/kg/day * 4). PMID- 21646485 TI - Impact of the E540V amino acid substitution in GyrB of Mycobacterium tuberculosis on quinolone resistance. AB - Amino acid substitutions conferring resistance to quinolones in Mycobacterium tuberculosis have generally been found within the quinolone resistance determining regions (QRDRs) in the A subunit of DNA gyrase (GyrA) rather than the B subunit of DNA gyrase (GyrB). To clarify the contribution of an amino acid substitution, E540V, in GyrB to quinolone resistance in M. tuberculosis, we expressed recombinant DNA gyrases in Escherichia coli and characterized them in vitro. Wild-type and GyrB-E540V DNA gyrases were reconstituted in vitro by mixing recombinant GyrA and GyrB. Correlation between the amino acid substitution and quinolone resistance was assessed by the ATP-dependent DNA supercoiling assay, quinolone-inhibited supercoiling assay, and DNA cleavage assay. The 50% inhibitory concentrations of eight quinolones against DNA gyrases bearing the E540V amino acid substitution in GyrB were 2.5- to 36-fold higher than those against the wild-type enzyme. Similarly, the 25% maximum DNA cleavage concentrations were 1.5- to 14-fold higher for the E540V gyrase than for the wild type enzyme. We further demonstrated that the E540V amino acid substitution influenced the interaction between DNA gyrase and the substituent(s) at R-7, R-8, or both in quinolone structures. This is the first detailed study of the contribution of the E540V amino acid substitution in GyrB to quinolone resistance in M. tuberculosis. PMID- 21646486 TI - Commonality among fluoroquinolone-resistant sequence type ST131 extraintestinal Escherichia coli isolates from humans and companion animals in Australia. AB - Escherichia coli sequence type 131 (ST131), an emergent multidrug-resistant extraintestinal pathogen, has spread epidemically among humans and was recently isolated from companion animals. To assess for human-companion animal commonality among ST131 isolates, 214 fluoroquinolone-resistant extraintestinal E. coli isolates (205 from humans, 9 from companion animals) from diagnostic laboratories in Australia, provisionally identified as ST131 by PCR, selectively underwent PCR based O typing and bla(CTX-M-15) detection. A subset then underwent multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE) analysis, extended virulence genotyping, antimicrobial susceptibility testing, and fluoroquinolone resistance genotyping. All isolates were O25b positive, except for two O16 isolates and one O157 isolate, which (along with six O25b-positive isolates) were confirmed by MLST to be ST131. Only 12% of isolates (25 human, 1 canine) exhibited bla(CTX-M-15). PFGE analysis of 20 randomly selected human and all 9 companion animal isolates showed multiple instances of >=94% profile similarity across host species; 12 isolates (6 human, 6 companion animal) represented pulsotype 968, the most prevalent ST131 pulsotype in North America (representing 23% of a large ST131 reference collection). Virulence gene and antimicrobial resistance profiles differed minimally, without host species specificity. The analyzed ST131 isolates also exhibited a conserved, host species independent pattern of chromosomal fluoroquinolone resistance mutations. However, eight (89%) companion animal isolates, versus two (10%) human isolates, possessed the plasmid-borne qnrB gene (P < 0.001). This extensive across-species strain commonality, plus the similarities between Australian and non-Australian ST131 isolates, suggest that ST131 isolates are exchanged between humans and companion animals both within Australia and intercontinentally. PMID- 21646487 TI - Pharmacokinetics, pharmacodynamics, and allometric scaling of chloroquine in a murine malaria model. AB - Chloroquine (CQ) is an important antimalarial drug for the treatment of special patient groups and as a comparator for preclinical testing of new drugs. Pharmacokinetic data for CQ in animal models are limited; thus, we conducted a three-part investigation, comprising (i) pharmacodynamic studies of CQ and CQ plus dihydroartemisinin (DHA) in Plasmodium berghei-infected mice, (ii) pharmacokinetic studies of CQ in healthy and malaria-infected mice, and (iii) interspecies allometric scaling for CQ from 6 animal and 12 human studies. The single-dose pharmacodynamic study (10 to 50 mg CQ/kg of body weight) showed dose related reduction in parasitemia (5- to >500-fold) and a nadir 2 days after the dose. Multiple-dose regimens (total dose, 50 mg/kg CQ) demonstrated a lower nadir and longer survival time than did the same single dose. The CQ-DHA combination provided an additive effect compared to each drug alone. The elimination half life (t(1/2)), clearance (CL), and volume of distribution (V) of CQ were 46.6 h, 9.9 liters/h/kg, and 667 liters/kg, respectively, in healthy mice and 99.3 h, 7.9 liters/h/kg, and 1,122 liters/kg, respectively, in malaria-infected mice. The allometric equations for CQ in healthy mammals (CL = 3.86 * W(0.56), V = 230 * W(0.94), and t(1/2) = 123 * W(0.2)) were similar to those for malaria-infected groups. CQ showed a delayed dose-response relationship in the murine malaria model and additive efficacy when combined with DHA. The biphasic pharmacokinetic profiles of CQ are similar across mammalian species, and scaling of specific parameters is plausible for preclinical investigations. PMID- 21646488 TI - Structural variations of staphylococcal cassette chromosome mec type IVa in Staphylococcus aureus clonal complex 8 and unrelated lineages. AB - PCR mapping of staphylococcal cassette chromosome mec type IVa and adjacent mobile elements in 94 methicillin-resistant Staphylococcus aureus (MRSA) strains identified two primary structures (A and B) that could be further classified into two (A1 and A2) and five (B1 to B5) variants, primarily based on structural differences in the orfX-J3 region. While spa type t008 (USA300) invariably contained the A variants, other spa types belonging to clonal complex 8 and unrelated lineages generally contained B variants. These findings have important implications for the typing and identification of MRSA strains containing B variants. PMID- 21646489 TI - Susceptibility of vertilmicin to modifications by three types of recombinant aminoglycoside-modifying enzymes. AB - The susceptibilities of vertilmicin and seven reference aminoglycosides to modifications by six recombinant aminoglycoside-modifying enzymes, AAC(6')-Ie, APH(2'')-Ia, AAC(6')-Ie-APH(2'')-Ia, ANT(2'')-Ia, AAC(6')-Ib, and AAC(6')-Ib-cr, were studied by coupled spectrophotometric assays in microtiter plates. In comparison to other aminoglycosides, the susceptibility of vertilmicin was 45.8- to 250.0-fold lower for AAC(6')-Ie acetylation, 39.2- to 116.7-fold lower for AAC(6')-Ie-APH(2'')-Ia acetylation, and 1.8- to 7.5-fold lower for ANT(2'')-Ia adenylation (except that shown by amikacin) while relatively comparable for AAC(6')-Ib acetylation, AAC(6')-Ib-cr acetylation, APH(2'')-Ia phosphorylation, and AAC(6')-Ie-APH(2'')-Ia phosphorylation. PMID- 21646490 TI - Emergence of NDM-1 metallo-beta-lactamase in Pseudomonas aeruginosa clinical isolates from Serbia. AB - This work reports, for the first time, the presence of New Delhi metallo-beta lactamase 1 (NDM-1) in Pseudomonas aeruginosa. Moreover, this is the first report of the NDM-1 presence in the Balkan region. Cosmid gene libraries of carbapenem nonsusceptible Pseudomonas aeruginosa clinical isolates MMA83 and MMA533 were screened for the presence of metallo-beta-lactamases. Accordingly, both MMA83 and MMA533 carried the bla(NDM-1) gene. Pulsed-field gel electrophoresis (PFGE) analysis indicated that strains MMA83 and MMA533 belonged to different clonal groups. Five additional isolates from different patients clonally related to either MMA83 or MMA533 were found to be NDM-1 positive. PMID- 21646491 TI - The expression levels of outer membrane proteins STM1530 and OmpD, which are influenced by the CpxAR and BaeSR two-component systems, play important roles in the ceftriaxone resistance of Salmonella enterica serovar Typhimurium. AB - Significant increases in STM3031, STM1530, and AcrD protein levels and significant decreases in OmpC and OmpD protein levels are present when the ceftriaxone-resistant Salmonella enterica serovar Typhimurium R200 strain is compared with the ceftriaxone-susceptible strain 01-4. AcrD is known to be involved in drug export, and STM3031 seems to play a key role in ceftriaxone resistance. Here, we examine the roles of STM1530, OmpC, and OmpD in ceftriaxone resistance. An ompD gene deletion mutant showed 4-fold higher ceftriaxone resistance than 01-4. An ompC gene deletion mutant showed 4-fold higher cephalothin and erythromycin resistance than 01-4, but there was no effect on ceftriaxone resistance. However, a stm1530 deletion mutant did show >64-fold lower ceftriaxone resistance than R200. Moreover, the STM3031 protein was significantly decreased in R200(Deltastm1530) compared to R200. STM3031 expression has been shown to be influenced by the two-component system regulator gene baeR. CpxR seems to modulate BaeR. A cpxA-cpxR gene deletion mutant showed >2,048-fold lower ceftriaxone resistance than R200. The outer membrane protein profile of R200(DeltacpxAR) showed significant decreases in STM3031 and STM1530 compared to R200, while OmpD had returned to the level found in 01-4. Furthermore, the stm3031, stm1530, and ompD mRNA levels were correlated with their protein expression levels in these strains, while decreases in the mRNA levels of the efflux pump acrB, acrD, and acrF genes were found in R200(DeltacpxAR). Findings similar to those for R200(DeltacpxAR) were found for R200(DeltabaeSR). These results, together with those for STM3031 and the fact that STM1530 is an outer membrane protein, suggest that STM1530 and OmpD are influenced by the CpxAR and BaeSR two-component systems and that this contributes to S. enterica serovar Typhimurium ceftriaxone resistance. PMID- 21646492 TI - Effect of ciprofloxacin concentration on the frequency and nature of resistant mutants selected from Pseudomonas aeruginosa mutS and mutT hypermutators. AB - The rapid emergence of drug resistance upon treatment of Pseudomonas aeruginosa infections with fluoroquinolones is a serious concern. In this study, we report the effect of hypermutability on the mutant selection window for ciprofloxacin (CIP) by comparing the hypermutator MPAO1 mutS and mutT strains with the wild type strain. The mutant selection window was shifted to higher CIP concentrations for both hypermutators, presenting the mutS strain with a broader selection window in comparison to the wild-type strain. The mutation prevention concentrations (MPC) determined for mutT and mutS strains were increased 2- and 4 fold over the wild-type level, respectively. In addition, we analyzed the molecular bases for resistance in the bacterial subpopulations selected at different points in the window. At the top of the window, the resistant clones isolated were mainly mutated in GyrA and ParC topoisomerase subunits, while at the bottom of the window, resistance was associated with the overexpression of MexCD-OprJ and MexAB-OprM efflux pumps. Accordingly, a greater proportion of multidrug-resistant clones were found among the subpopulations isolated at the lower CIP concentrations. Furthermore, we found that the exposure to CIP subinhibitory concentrations favors the accumulation of cells overexpressing MexCD-OprJ (due to mutations in the transcriptional repressor NfxB) and MexAB OprM efflux pumps. We discuss these results in the context of the possible participation of this antibiotic in a mutagenic process. PMID- 21646493 TI - ACCF/AHA 2011 key data elements and definitions of a base cardiovascular vocabulary for electronic health records: a report of the American College of Cardiology Foundation/American Heart Association Task Force on Clinical Data Standards. PMID- 21646494 TI - Optical coherence tomographic analysis of in-stent neoatherosclerosis after drug eluting stent implantation. AB - BACKGROUND: We report findings from optical coherence tomography (OCT) of in stent neoatherosclerosis as a cause of drug-eluting stent (DES) failure. METHODS AND RESULTS: Optical coherence tomography and grayscale and virtual histology intravascular ultrasound were performed in 50 patients (30 stable, 20 unstable angina) with 50 DES in-stent restenosis lesions and intimal hyperplasia >50% of stent area. Median follow-up time was 32.2 months. Overall, 26 lesions (52%) had at least 1 OCT-defined in-stent thin-cap fibroatheroma (TCFA)-containing neointima and 29 (58%) had at least 1 in-stent neointimal rupture. Patients presenting with unstable angina showed a thinner fibrous cap (55 MUm [interquartile range 42 to 105 MUm] versus 100 MUm [interquartile range 60 to 205 MUm], P=0.006) and higher incidence of OCT-defined TCFA-containing neointima (75% versus 37%, P=0.008), intimal rupture (75% versus 47%, P=0.044), thrombi (80% versus 43%, P=0.010), and red thrombi (30% versus 3%, P=0.012) than stable patients. Fibrous cap thickness negatively correlated with follow-up time (r= 0.318, P=0.024). Compared with DES <20 months after implantation (the best cut off to predict TCFA-containing neointima), DES >=20 months after implantation had a higher incidence of TCFA-containing neointima (69% versus 33%, P=0.012) and red thrombi (27% versus 0%, P=0.007). Patients with unstable (versus stable) angina had an increasing number of unstable OCT findings including TCFA-containing neointima, neointima rupture, and thrombus (P=0.027). The rate of agreement between grayscale intravascular ultrasound and OCT for detecting intimal rupture was 50% and for detecting thrombus was 44%. The agreement between virtual histology intravascular ultrasound and OCT for identifying TCFA-containing neointima was 78%. CONCLUSIONS: In-stent neoatherosclerosis may be an important mechanism of DES failure, especially late after implantation. PMID- 21646495 TI - Incidence and prognostic value of early repolarization pattern in the 12-lead electrocardiogram. AB - BACKGROUND: Early repolarization pattern is a common ECG finding characterized by J-point elevation and QRS notching or slurring in the inferior and/or lateral leads, yet little is known about its incidence and long-term prognosis in Asian populations. METHODS AND RESULTS: We reviewed all the ECG records of the 5976 atomic-bomb survivors who were examined at least once during our biennial health examination in Nagasaki, Japan, between July 1958 and December 2004. We defined early repolarization pattern as >=0.1-mV elevation of the J point or ST segment, with notching or slurring in at least 2 inferior and/or lateral leads. We assessed unexpected, cardiac, and all-cause death risk by Cox analysis. We identified 1429 early repolarization pattern cases (779 incident cases) during follow-up, yielding a positive rate of 23.9% and an incidence rate of 715 per 100 000 person-years. Early repolarization pattern had an elevated risk of unexpected death (hazard ratio, 1.83; 95% confidence interval, 1.12 to 2.97; P=0.02) and a decreased risk of cardiac (hazard ratio, 0.75; 95% confidence interval, 0.60 to 0.93; P<0.01) and all-cause (hazard ratio, 0.85; 95% confidence interval, 0.78 to 0.93; P<0.01) death. In addition, both slurring and notching were related to higher risk of unexpected death (hazard ratio, 2.09; 95% confidence interval, 1.06 to 4.12; P=0.03), as was early repolarization pattern manifestation in both inferior and lateral leads (hazard ratio, 2.50; 95% confidence interval, 1.29 to 4.83; P<0.01). CONCLUSIONS: Early repolarization pattern is associated with an elevated risk of unexpected death and a decreased risk of cardiac and all-cause death. Specific early repolarization pattern morphologies and location are associated with an adverse prognosis. PMID- 21646496 TI - Chronic kidney disease is associated with the incidence of atrial fibrillation: the Atherosclerosis Risk in Communities (ARIC) study. AB - BACKGROUND: Chronic kidney disease is associated with the incidence of cardiovascular disease. Chronic kidney disease may also increase the risk of atrial fibrillation (AF), but existing studies have reported inconsistent results. METHODS AND RESULTS: We estimated cystatin C-based glomerular filtration rate (eGFR(cys)) and measured urinary albumin-to-creatinine ratio (ACR) in 10 328 men and women free of AF from the Atherosclerosis Risk in Communities (ARIC) Study in 1996 to 1998. Incidence of AF was ascertained through the end of 2007. During a median follow-up of 10.1 years, we identified 788 incident AF cases. Compared with individuals with eGFR(cys) >=90 mL . min(-1) . 1.73 m(-2), multivariable hazard ratios and 95% confidence intervals (CIs) of AF were 1.3 (95% CI, 1.1 to 1.6), 1.6 (95% CI, 1.3 to 2.1), and 3.2 (95% CI, 2.0 to 5.0; P for trend <0.0001) in those with eGFR(cys) of 60 to 89, 30 to 59, and 15 to 29 mL . min(-1) . 1.73 m(-2), respectively. Similarly, the presence of macroalbuminuria (ACR >=300 mg/g; hazard ratio, 3.2; 95% CI, 2.3 to 4.5) and microalbuminuria (ACR, 30 to 299 mg/g; hazard ratio, 2.0; 95% CI, 1.6 to 2.4) was associated with higher AF risk compared with those with ACR <30 mg/g. Risk of AF was particularly elevated in those with both low eGFR(cys) and macroalbuminuria (hazard ratio, 13.1; 95% CI, 6.0 to 28.6, comparing individuals with ACR >=300 mg/g and eGFR(cys) of 15 to 29 mL . min(-1) . 1.73 m(-2) and those with ACR <30 mg/g and eGFR(cys) >=90 mL . min(-1) . 1.73 m(-2)). CONCLUSION: In this large population based study, reduced kidney function and presence of albuminuria were strongly associated with the incidence of AF independently of other risk factors. PMID- 21646497 TI - Five-year outcomes of sirolimus-eluting and paclitaxel-eluting stents in routine clinical practice: are they equally good (or bad)? PMID- 21646498 TI - Repolarization alternans reveals vulnerability to human atrial fibrillation. AB - BACKGROUND: The substrates for human atrial fibrillation (AF) are poorly understood, but involve abnormal repolarization (action potential duration [APD]). We hypothesized that beat-to-beat oscillations in APD may explain AF substrates, and why vulnerability to AF forms a spectrum from control subjects without AF to patients with paroxysmal then persistent AF. METHODS AND RESULTS: In 33 subjects (12 with persistent AF, 13 with paroxysmal AF, and 8 controls without AF), we recorded left (n=33) and right (n=6) atrial APD on pacing from cycle lengths 600 to 500 ms (100 to 120 bpm) up to the point where AF initiated. Action potential duration alternans required progressively faster rates for patients with persistent AF, patients with paroxysmal AF, and controls (cycle length 411+/-94 versus 372+/-72 versus 218+/-33 ms; P<0.01). In AF patients, APD alternans occurred at rates as slow as 100 to 120 bpm, unrelated to APD restitution (P>0.10). In this milieu, spontaneous ectopy initiated AF. At fast rates, APD alternans disorganized to complex oscillations en route to AF. Complex oscillations also arose at progressively faster rates for persistent AF, paroxysmal AF, and controls (cycle length: 316+/-99 versus 266+/-19 versus 177+/ 16 ms; P=0.02). In paroxysmal AF, APD oscillations amplified before AF (P<0.001). In controls, APD alternans arose only at very fast rates (cycle length <250 ms; P<0.001 versus AF groups) just preceding AF. In 4 AF patients in whom rapid pacing did not initiate AF, APD alternans arose transiently then extinguished. CONCLUSIONS: Atrial APD alternans reveals dynamic substrates for AF, arising most readily (at lower rates and higher magnitudes) in persistent AF then paroxysmal AF, and least readily in controls. APD alternans preceded all AF episodes and was absent when AF did not initiate. The cellular mechanisms for APD alternans near resting heart rates require definition. PMID- 21646499 TI - Late outcomes of a single-center experience of 400 consecutive thoracic endovascular aortic repairs. AB - BACKGROUND: In this study, we report the late outcomes of a large, decade-long single-center thoracic endovascular aortic repair experience. METHODS AND RESULTS: A prospectively maintained registry and the electronic medical records of 400 consecutive thoracic endovascular aortic repair performed at a tertiary care center were reviewed. The distribution of pathologies treated included aneurysms (198, 49%), dissections (100, 25%), penetrating ulcers (54, 14%), traumatic transections (25, 6%), and other pathologies (23, 6%). Spinal drains were placed prophylactically in 127 cases (32%) of planned extended aortic coverage. There were no acute surgical conversions. Adjunctive surgical procedures were performed on 94 patients (24%). Subclavian revascularizations were performed selectively in only 15% of zone 0 to 2 deployments. The median length of stay was 5 days (limits, 1 and 79 days). Overall 30-day mortality was 6.5% (elective, 2.6%; urgent, 9.5%; and emergent, 20%). Permanent spinal cord ischemia occurred in 4.5% and stroke in 3%. Kaplan-Meier estimates of survival were 82%, 76%, 68%, and 60% and freedom from secondary intervention was 90%, 86%, 81%, and 78% at 6, 12, 24, and 36 months, respectively. Risk factors for mortality included stroke, urgent/emergent repair, age >=80 years, general anesthesia, and dissection pathology. CONCLUSIONS: Thoracic endovascular aortic repair may be used to treat a variety of thoracic aortic pathologies with a very low risk of intraoperative conversion. Overall rates of mortality and neurological complications were relatively low but significantly increased in emergent repairs. There appeared to be a substantial number of late deaths, which may represent a combination of poor patient selection and treatment failures. PMID- 21646500 TI - Five-year clinical and angiographic outcomes of a randomized comparison of sirolimus-eluting and paclitaxel-eluting stents: results of the Sirolimus-Eluting Versus Paclitaxel-Eluting Stents for Coronary Revascularization LATE trial. AB - BACKGROUND: Long-term comparative data of first-generation drug-eluting stents are scarce. We investigated clinical and angiographic outcomes of sirolimus eluting (SES) and paclitaxel-eluting stents (PES) at 5 years as part of the Sirolimus-Eluting Versus Paclitaxel-Eluting Stents for Coronary Revascularization (SIRTAX) LATE study. METHODS AND RESULTS: A total of 1012 patients were randomly assigned to SES or PES. Repeat angiography was completed in 444 of 1012 patients (43.8%) at 5 years. Major adverse cardiac events occurred in 19.7% of SES- and 21.4% of PES-treated patients (hazard ratio, 0.89; 95% confidence interval, 0.68 to 1.17; P=0.39) at 5 years. There were no differences between SES and PES in terms of cardiac death (5.8% versus 5.7%; P=0.35), myocardial infarction (6.6% versus 6.9%; P=0.51), and target lesion revascularization (13.1% versus 15.1%; P=0.29). Between 1 and 5 years, the annual rate of target lesion revascularization was 2.0% (95% confidence interval, 1.4% to 2.6%) for SES and 1.4% (95% confidence interval, 0.9% to 2.0%) for PES. Among patients undergoing paired angiography at 8 months and 5 years, delayed lumen loss amounted to 0.37 +/- 0.73 mm for SES and 0.29 +/- 0.59 mm for PES (P=0.32). The overall rate of definite stent thrombosis was 4.6% for SES and 4.1% for PES (P=0.74), and very late definite stent thrombosis occurred at an annual rate of 0.65% (95% confidence interval, 0.40% to 0.90%). CONCLUSIONS: Long-term follow-up of first generation drug-eluting stents shows no significant differences in clinical and angiographic outcomes between SES and PES. The continuous increase in late lumen loss in conjunction with the ongoing risk of very late stent thrombosis suggests that vascular healing remains incomplete up to 5 years after implantation of first-generation drug-eluting stents. PMID- 21646501 TI - Targeting viable myocardium in cardiac resynchronization therapy using a multipolar left ventricular lead. PMID- 21646502 TI - Letter by Murzi and Glauber regarding article, "Extensive primary repair of the thoracic aorta in acute type A aortic dissection by means of ascending aorta replacement combined with open placement of triple-branched stent graft: early results". PMID- 21646503 TI - Letter by Ferdinand et al regarding article, "Does black ethnicity influence the development of stent thrombosis in the drug-eluting stent era?". PMID- 21646504 TI - Letter by Smith et al regarding article, "Does black ethnicity influence the development of stent thrombosis in the drug-eluting stent era?". PMID- 21646505 TI - Sex and race differences in right ventricular structure and function: the multi ethnic study of atherosclerosis-right ventricle study. AB - BACKGROUND: Right ventricular (RV) morphology is an important predictor of outcomes in heart and lung disease; however, determinants of RV anatomy have not been well studied. We examined the demographic factors associated with RV morphology and function in a population-based multiethnic sample free of clinical cardiovascular disease. METHODS AND RESULTS: In the Multi-Ethnic Study of Atherosclerosis (MESA), cardiac magnetic resonance imaging was performed on 5098 participants. Right ventricular volumes and mass were available for 4204 participants. Normative equations for RV parameters were derived with an allometric approach. The study sample (n=4123) was 61.5+/-10.1 years of age and 47.5% men. Older age was associated with lower RV mass (~5% lower mass per decade), with larger age-related decrements in men than in women (P<0.05 for interaction). Older age was also associated with higher RV ejection fraction, an association that differed between races/ethnicities (P<=0.01 for interaction). Overall, men had greater RV mass (~8%) and larger RV volumes than women, but had lower RV ejection fraction (4% in absolute terms; P<0.001). Blacks had lower RV mass than whites (P<=0.002), whereas Hispanics had higher RV mass (P<=0.02). When the derived normative equations were used, 7.3% (95% confidence interval, 6.5 to 8.1) met the criteria for RV hypertrophy, and 5.9% (95% confidence interval, 5.2 to 6.6) had RV dysfunction. CONCLUSION: Age, sex, and race are associated with significant differences in RV mass, RV volumes, and RV ejection fraction, potentially explaining distinct responses of the RV to cardiopulmonary disease. PMID- 21646506 TI - Current status of endovascular stroke treatment. AB - The management of acute ischemic stroke is rapidly developing.Although acute ischemic stroke is a major cause of adult disability and death, the number of patients requiring emergency endovascular intervention remains unknown, but is a fraction of the overall stroke population. Public health initiatives endeavor to raise public awareness about acute stroke to improve triage for emergency treatment, and the medical community is working to develop stroke services at community and academic medical centers throughout the United States. There is an Accreditation Council for Graduate Medical Education-approved pathway for training in endovascular surgical neuroradiology, the specialty designed to train physicians specifically to treat cerebrovascular diseases. Primary and comprehensive stroke center designations have been defined, yet questions remain about the best delivery model. Telemedicine is available to help community medical centers cope with the complexity of stroke triage and treatment. Should comprehensive care be provided at every community center, or should patients with complex medical needs be triaged to major stroke centers with high-level surgical,intensive care, and endovascular capabilities? Although the answers to these and other questions about stroke care delivery remain unanswered owing to the paucity of empirical data, we are convinced that stroke care regionalization is crucial for delivery of high-quality comprehensive ischemic stroke treatment. A stroke team available 24 hours per day, 7 days per week requires specialty skills in stroke neurology, endovascular surgical neuroradiology, neurosurgery, neurointensive care, anesthesiology, nursing, and technical support for optimal success. Several physician groups with divergent training backgrounds (i.e., interventional neuroradiology, neurosurgery,neurology, peripheral interventional radiology, and cardiology) lay claim to the treatment of stroke patients,particularly the endovascular or interventional methods. Few would challenge neurologists over the responsibility for emergency evaluation and triage of stroke victims for intra intravenous fibrinolysis, even though emergency physicians are most commonly the first to evaluate these patients. There are many unanswered questions about the role of imaging in defining best treatment. Perfusion imaging with CT or MRI appears to have relevance even though its role remains undefined and is the subject of ongoing research. Meanwhile, investigators are exploring new, and perhaps more specific,imaging methods with cerebral metabolic rate of oxygen and cellular acid-base imbalance. There are currently 6 ongoing trials of stroke intervention, many with proprietary technologies and private funding, competing for the same patient population as multicenter trials funded by the NIH. At the same time, much of the interventional stroke treatment currently occurs outside of trials in the community and academic settings without the collection of much-needed data. Market forces will certainly shape future stroke therapy, but it is unclear whether the current combination of private and public funding for these endeavors is the best method of development. PMID- 21646507 TI - Prosthetic heart valve. PMID- 21646508 TI - A painful fistula. PMID- 21646509 TI - Achieving ideal cardiovascular and brain health: opportunity amid crisis: Presidential Address at the American Heart Association 2010 Scientific Sessions. PMID- 21646511 TI - Sequential and spatially restricted interactions of assembly factors with an autotransporter beta domain. AB - Autotransporters are bacterial virulence factors that consist of an N-terminal extracellular ("passenger") domain and a C-terminal beta barrel domain ("beta domain") that resides in the outer membrane. Here we used an in vivo site specific photocrosslinking approach to gain insight into the mechanism by which the beta domain is integrated into the outer membrane and the relationship between beta domain assembly and passenger domain secretion. We found that periplasmic chaperones and specific components of the beta barrel assembly machinery (Bam) complex interact with the beta domain of the Escherichia coli O157:H7 autotransporter extracellular serine protease P (EspP) in a temporally and spatially regulated fashion. Although the chaperone Skp initially interacted with the entire beta domain, BamA, BamB, and BamD subsequently interacted with discrete beta domain regions. BamB and BamD remained bound to the beta domain longer than BamA and therefore appeared to function at a later stage of assembly. Interestingly, we obtained evidence that the completion of beta domain assembly is regulated by an intrinsic checkpoint mechanism that requires the completion of passenger domain secretion. In addition to leading to a detailed model of autotransporter biogenesis, our results suggest that the lipoprotein components of the Bam complex play a direct role in the membrane integration of beta barrel proteins. PMID- 21646512 TI - Loss of Bardet-Biedl syndrome protein-8 (BBS8) perturbs olfactory function, protein localization, and axon targeting. AB - Bardet-Biedl syndrome (BBS) is a pleiotropic, heterogeneous human disease whose etiology lies primarily in dysfunctional basal bodies and/or cilia. Both BBS patients and several BBS mouse models exhibit impaired olfactory function. To explore the nature of olfactory defects in BBS, a genetic ablation of the mouse Bbs8 gene that incorporates a fluorescent reporter protein was created. The endogenous BBS8 protein and reporter are particularly abundant in olfactory sensory neurons (OSNs), and specific BBS8 antibodies reveal staining in the dendritic knob in a shell-like structure that surrounds the basal bodies. Bbs8 null mice have reduced olfactory responses to a number of odorants, and immunohistochemical analyses reveal a near-complete loss of cilia from OSNs and mislocalization of proteins normally enriched in cilia. To visualize altered protein localization in OSNs, we generated a SLP3(eGFP) knock-in mouse and imaged the apical epithelium, including dendritic knobs and proximal cilia, in ex vivo tissue preparations. Additionally, protein reagents that reflect the characteristic neuronal activity of each OSN revealed altered activity in Bbs8 null cells. In addition to previously known defects at the ciliary border, we also observed aberrant targeting of OSN axons to the olfactory bulb; axons expressing the same receptor display reduced fasciculation and project to multiple targets in the olfactory bulb. We suggest that loss of BBS8 leads to a dramatic and variable reduction in cilia, the essential signaling platform for olfaction, which alters the uniformity of responses in populations of OSNs expressing the same receptor, thereby contributing to the observed axon-targeting defects. PMID- 21646513 TI - Human cystic fibrosis airway epithelia have reduced Cl- conductance but not increased Na+ conductance. AB - Loss of cystic fibrosis transmembrane conductance regulator (CFTR) anion channel function causes cystic fibrosis (CF) lung disease. CFTR is expressed in airway epithelia, but how CF alters electrolyte transport across airway epithelia has remained uncertain. Recent studies of a porcine model showed that in vivo, excised, and cultured CFTR(-/-) and CFTR(DeltaF508/DeltaF508) airway epithelia lacked anion conductance, and they did not hyperabsorb Na(+). Therefore, we asked whether Cl(-) and Na(+) conductances were altered in human CF airway epithelia. We studied differentiated primary cultures of tracheal/bronchial epithelia and found that transepithelial conductance (Gt) under basal conditions and the cAMP stimulated increase in Gt were markedly attenuated in CF epithelia compared with non-CF epithelia. These data reflect loss of the CFTR anion conductance. In CF and non-CF epithelia, the Na(+) channel inhibitor amiloride produced similar reductions in Gt and Na(+) absorption, indicating that Na(+) conductance in CF epithelia did not exceed that in non-CF epithelia. Consistent with previous reports, adding amiloride caused greater reductions in transepithelial voltage and short-circuit current in CF epithelia than in non-CF epithelia; these changes are attributed to loss of a Cl(-) conductance. These results indicate that Na(+) conductance was not increased in these cultured CF tracheal/bronchial epithelia and point to loss of anion transport as key to airway epithelial dysfunction in CF. PMID- 21646514 TI - Explaining complex codon usage patterns with selection for translational efficiency, mutation bias, and genetic drift. AB - The genetic code is redundant with most amino acids using multiple codons. In many organisms, codon usage is biased toward particular codons. Understanding the adaptive and nonadaptive forces driving the evolution of codon usage bias (CUB) has been an area of intense focus and debate in the fields of molecular and evolutionary biology. However, their relative importance in shaping genomic patterns of CUB remains unsolved. Using a nested model of protein translation and population genetics, we show that observed gene level variation of CUB in Saccharomyces cerevisiae can be explained almost entirely by selection for efficient ribosomal usage, genetic drift, and biased mutation. The correlation between observed codon counts within individual genes and our model predictions is 0.96. Although a variety of factors shape patterns of CUB at the level of individual sites within genes, our results suggest that selection for efficient ribosome usage is a central force in shaping codon usage at the genomic scale. In addition, our model allows direct estimation of codon-specific mutation rates and elongation times and can be readily applied to any organism with high-throughput expression datasets. More generally, we have developed a natural framework for integrating models of molecular processes to population genetics models to quantitatively estimate parameters underlying fundamental biological processes, such a protein translation. PMID- 21646515 TI - Direct conversion of human fibroblasts to dopaminergic neurons. AB - Recent reports demonstrate that somatic mouse cells can be directly converted to other mature cell types by using combined expression of defined factors. Here we show that the same strategy can be applied to human embryonic and postnatal fibroblasts. By overexpression of the transcription factors Ascl1, Brn2, and Myt1l, human fibroblasts were efficiently converted to functional neurons. We also demonstrate that the converted neurons can be directed toward distinct functional neurotransmitter phenotypes when the appropriate transcriptional cues are provided together with the three conversion factors. By combining expression of the three conversion factors with expression of two genes involved in dopamine neuron generation, Lmx1a and FoxA2, we could direct the phenotype of the converted cells toward dopaminergic neurons. Such subtype-specific induced neurons derived from human somatic cells could be valuable for disease modeling and cell replacement therapy. PMID- 21646516 TI - Host and viral ecology determine bat rabies seasonality and maintenance. AB - Rabies is an acute viral infection that is typically fatal. Most rabies modeling has focused on disease dynamics and control within terrestrial mammals (e.g., raccoons and foxes). As such, rabies in bats has been largely neglected until recently. Because bats have been implicated as natural reservoirs for several emerging zoonotic viruses, including SARS-like corona viruses, henipaviruses, and lyssaviruses, understanding how pathogens are maintained within a population becomes vital. Unfortunately, little is known about maintenance mechanisms for any pathogen in bat populations. We present a mathematical model parameterized with unique data from an extensive study of rabies in a Colorado population of big brown bats (Eptesicus fuscus) to elucidate general maintenance mechanisms. We propose that life history patterns of many species of temperate-zone bats, coupled with sufficiently long incubation periods, allows for rabies virus maintenance. Seasonal variability in bat mortality rates, specifically low mortality during hibernation, allows long-term bat population viability. Within viable bat populations, sufficiently long incubation periods allow enough infected individuals to enter hibernation and survive until the following year, and hence avoid an epizootic fadeout of rabies virus. We hypothesize that the slowing effects of hibernation on metabolic and viral activity maintains infected individuals and their pathogens until susceptibles from the annual birth pulse become infected and continue the cycle. This research provides a context to explore similar host ecology and viral dynamics that may explain seasonal patterns and maintenance of other bat-borne diseases. PMID- 21646517 TI - Maturation time of new granule cells in the dentate gyrus of adult macaque monkeys exceeds six months. AB - We studied two groups of adult macaque monkeys to determine the time course of adult neurogenesis in the dentate gyrus of the hippocampus. In the first group, six adult monkeys (Macaca mulatta) received a single injection of the thymidine analog BrdU (75 mg/kg), which is incorporated into replicating DNA and serves as a marker for new cell birth. Brain tissue was collected 48 h, 2 wk, and 6 wk after BrdU injection to examine the initial stages of neurogenesis. Because mature neurons were not evident at 6 wk, we examined tissue collected over a longer time course in a second study. In this study, eight monkeys (Macaca fascicularis) who were subjects in a separate exercise study received 10 weekly injections of BrdU (75 mg/kg), and brain tissue was collected at 16 and 28 wk from the first injection. Based on the timing of expression of neuronal cell markers (betaIII-tubulin, doublecortin, NeuN), the extent of dendritic arborization, and acquisition of mature cell body morphology, we show that granule cell maturation in the dentate gyrus of a nonhuman primate is protracted over a minimum of a 6-mo time period, more than 6 times longer than in rodents. The lengthened time course for new cell maturation in nonhuman primates may be appropriate for preservation of neural plasticity over their longer life span and is relevant to our understanding of antidepressant and other therapies that have been linked to neurogenesis in humans. PMID- 21646518 TI - Generation of a unique small molecule peptidomimetic that neutralizes lupus autoantibody activity. AB - Systemic lupus erythematosus (SLE) is a chronic autoimmune disease characterized by the presence of pathogenic autoantibodies, many of which are directed against nuclear antigens, in particular double-stranded (ds) DNA. Both clinical studies and animal models have shown that anti-dsDNA antibodies contribute to kidney disease, which is present in 50% of lupus patients and is a major cause of mortality. We previously demonstrated that a subset of nephrotoxic anti-dsDNA antibodies also recognizes the pentapeptide consensus sequence D/E W D/E Y S/G (DWEYS) present in the NR2A and NR2B subunits of the N-methyl-d-aspartate receptor (NMDAR). Autoantibodies with this specificity are present in ~40% of lupus patient sera and are both nephrotoxic and neurotoxic. Elevated titers are present in cerebrospinal fluid of patients with central nervous system manifestations of SLE. Administration of the nonnaturally occurring D form of the DWEYS pentapeptide prevents these antibodies from depositing in glomeruli and from mediating neuronal excitotoxicity. To craft a more useful therapeutic, we used the structural features of the DWEYS peptide to design a unique, selective, and potent small molecule peptidomimetic, FISLE-412, which neutralizes anti dsDNA/NMDAR lupus autoantibodies and prevents their pathogenic interaction with tissue antigens. This compound, or others derived from it, may provide a unique strategy for the development of lupus therapeutics. PMID- 21646519 TI - Preexisting drug-resistance mutations reveal unique barriers to resistance for distinct antivirals. AB - Clinical trials of direct-acting antiviral agents in patients chronically infected with hepatitis C virus (HCV) have demonstrated that viral resistance is detected rapidly during monotherapy. In patients, HCV does not exist as a single, genetically homogenous virus but rather as a population of variants termed "quasispecies." Preexisting variants resistant to specific antiviral drugs, overlooked in traditional hit-to-lead discovery efforts, may be responsible for these poor clinical outcomes. To enable real-time studies of resistance emergence in live cells, we established fluorescent protein-labeled HCV replicon cell lines. We validated these cell lines by demonstrating that antiviral susceptibility and the selection of signature resistance mutations for various drug classes are similar to traditional replicon cell lines. By quantifying the kinetics and uniformity of replication within colonies of drug-resistant fluorescent replicon cells, we showed that resistance emerged from a single cell and preexisted in a treatment-naive replicon population. Within this population, we determined the relative frequency of preexisting replicons capable of establishing foci during treatment with distinct antivirals. By measuring relative frequency as a function of dose, we quantitatively ranked distinct antiviral molecules on the basis of their distinct barriers to resistance. These insights into RNA virus quasispecies structure provide guidance for selecting clinical drug concentrations and selecting antiviral drug combinations most likely to suppress resistance. PMID- 21646520 TI - Reference-guided assembly of four diverse Arabidopsis thaliana genomes. AB - We present whole-genome assemblies of four divergent Arabidopsis thaliana strains that complement the 125-Mb reference genome sequence released a decade ago. Using a newly developed reference-guided approach, we assembled large contigs from 9 to 42 Gb of Illumina short-read data from the Landsberg erecta (Ler-1), C24, Bur-0, and Kro-0 strains, which have been sequenced as part of the 1,001 Genomes Project for this species. Using alignments against the reference sequence, we first reduced the complexity of the de novo assembly and later integrated reads without similarity to the reference sequence. As an example, half of the noncentromeric C24 genome was covered by scaffolds that are longer than 260 kb, with a maximum of 2.2 Mb. Moreover, over 96% of the reference genome was covered by the reference-guided assembly, compared with only 87% with a complete de novo assembly. Comparisons with 2 Mb of dideoxy sequence reveal that the per-base error rate of the reference-guided assemblies was below 1 in 10,000. Our assemblies provide a detailed, genomewide picture of large-scale differences between A. thaliana individuals, most of which are difficult to access with alignment-consensus methods only. We demonstrate their practical relevance in studying the expression differences of polymorphic genes and show how the analysis of sRNA sequencing data can lead to erroneous conclusions if aligned against the reference genome alone. Genome assemblies, raw reads, and further information are accessible through http://1001genomes.org/projects/assemblies.html. PMID- 21646521 TI - Earliest human occupations at Dmanisi (Georgian Caucasus) dated to 1.85-1.78 Ma. AB - The early Pleistocene colonization of temperate Eurasia by Homo erectus was not only a significant biogeographic event but also a major evolutionary threshold. Dmanisi's rich collection of hominin fossils, revealing a population that was small-brained with both primitive and derived skeletal traits, has been dated to the earliest Upper Matuyama chron (ca. 1.77 Ma). Here we present archaeological and geologic evidence that push back Dmanisi's first occupations to shortly after 1.85 Ma and document repeated use of the site over the last half of the Olduvai subchron, 1.85-1.78 Ma. These discoveries show that the southern Caucasus was occupied repeatedly before Dmanisi's hominin fossil assemblage accumulated, strengthening the probability that this was part of a core area for the colonization of Eurasia. The secure age for Dmanisi's first occupations reveals that Eurasia was probably occupied before Homo erectus appears in the East African fossil record. PMID- 21646522 TI - Constraints to addressing food insecurity in protracted crises. AB - A substantial portion of the world's people have not made adequate progress toward overcoming hunger or achieving sustainable livelihoods. The classic approach to addressing chronic food insecurity has been a strategy of agricultural development, supplemented by humanitarian assistance in the event of a shock or crisis--an approach predicated on assumptions that do not fit the context of protracted crises. This article describes protracted crises and argues that they are sufficiently different to warrant special consideration, but there are unique constraints to engagement in protracted crises. The article explores the constraints promoting sustainable livelihoods in these contexts and proposes elements of an alternative approach. It evaluates the limited evidence available about such an approach and outlines important questions for further research. PMID- 21646523 TI - Nonlinear effect of climate on plague during the third pandemic in China. AB - Over the years, plague has caused a large number of deaths worldwide and subsequently changed history, not the least during the period of the Black Death. Of the three plague pandemics, the third is believed to have originated in China. Using the spatial and temporal human plague records in China from 1850 to 1964, we investigated the association of human plague intensity (plague cases per year) with proxy data on climate condition (specifically an index for dryness/wetness). Our modeling analysis demonstrates that the responses of plague intensity to dry/wet conditions were different in northern and southern China. In northern China, plague intensity generally increased when wetness increased, for both the current and the previous year, except for low intensity during extremely wet conditions in the current year (reflecting a dome-shaped response to current-year dryness/wetness). In southern China, plague intensity generally decreased when wetness increased, except for high intensity during extremely wet conditions of the current year. These opposite effects are likely related to the different climates and rodent communities in the two parts of China: In northern China (arid climate), rodents are expected to respond positively to high precipitation, whereas in southern China (humid climate), high precipitation is likely to have a negative effect. Our results suggest that associations between human plague intensity and precipitation are nonlinear: positive in dry conditions, but negative in wet conditions. PMID- 21646524 TI - Value-driven attentional capture. AB - Attention selects which aspects of sensory input are brought to awareness. To promote survival and well-being, attention prioritizes stimuli both voluntarily, according to context-specific goals (e.g., searching for car keys), and involuntarily, through attentional capture driven by physical salience (e.g., looking toward a sudden noise). Valuable stimuli strongly modulate voluntary attention allocation, but there is little evidence that high-value but contextually irrelevant stimuli capture attention as a consequence of reward learning. Here we show that visual search for a salient target is slowed by the presence of an inconspicuous, task-irrelevant item that was previously associated with monetary reward during a brief training session. Thus, arbitrary and otherwise neutral stimuli imbued with value via associative learning capture attention powerfully and persistently during extinction, independently of goals and salience. Vulnerability to such value-driven attentional capture covaries across individuals with working memory capacity and trait impulsivity. This unique form of attentional capture may provide a useful model for investigating failures of cognitive control in clinical syndromes in which value assigned to stimuli conflicts with behavioral goals (e.g., addiction, obesity). PMID- 21646525 TI - On-off system for PI3-kinase-Akt signaling through S-nitrosylation of phosphatase with sequence homology to tensin (PTEN). AB - Nitric oxide (NO) physiologically regulates numerous cellular responses through S nitrosylation of protein cysteine residues. We performed antibody-array screening in conjunction with biotin-switch assays to look for S-nitrosylated proteins. Using this combination of techniques, we found that phosphatase with sequence homology to tensin (PTEN) is selectively S-nitrosylated by low concentrations of NO at a specific cysteine residue (Cys-83). S-nitrosylation of PTEN (forming SNO PTEN) inhibits enzymatic activity and consequently stimulates the downstream Akt cascade, indicating that Cys-83 is a critical site for redox regulation of PTEN function. In ischemic mouse brain, we observed SNO-PTEN in the core and penumbra regions but found SNO-Akt, which is known to inhibit Akt activity, only in the ischemic core. These findings suggest that low concentrations of NO, as found in the penumbra, preferentially S-nitrosylate PTEN, whereas higher concentrations of NO, known to exist in the ischemic core, also S-nitrosylate Akt. In the penumbra, inhibition of PTEN (but not Akt) activity by S-nitrosylation would be expected to contribute to cell survival by means of enhanced Akt signaling. In contrast, in the ischemic core, SNO-Akt formation would inhibit this neuroprotective pathway. In vitro model systems support this notion. Thus, we identify unique sites of PTEN and Akt regulation by means of S-nitrosylation, resulting in an "on-off" pattern of control of Akt signaling. PMID- 21646526 TI - Domain expertise insulates against judgment bias by monetary favors through a modulation of ventromedial prefrontal cortex. AB - Recent work using an art-viewing paradigm shows that monetary sponsorship of the experiment by a company (a favor) increases the valuation of paintings placed next to the sponsoring corporate logo, an effect that correlates with modulation of the ventromedial prefrontal cortex (VMPFC). We used the same art-viewing paradigm to test a prevailing idea in the domain of conflict-of-interest: that expertise in a domain insulates against judgment bias even in the presence of a monetary favor. Using a cohort of art experts, we show that monetary favors do not bias the experts' valuation of art, an effect that correlates with a lack of modulation of the VMPFC across sponsorship conditions. The lack of sponsorship effect in the VMPFC suggests the hypothesis that their brains remove the behavioral sponsorship effect by censoring sponsorship-dependent modulation of VMPFC activity. We tested the hypothesis that prefrontal regions play a regulatory role in mediating the sponsorship effect. We show that the dorsolateral prefrontal cortex (DLPFC) is recruited in the expert group. Furthermore, we tested the hypothesis in nonexpert controls by contrasting brain responses in controls who did not show a sponsorship effect to controls who did. Changes in effective connectivity between the DLPFC and VMPFC were greater in nonexpert controls, with an absence of the sponsorship effect relative to those with a presence of the sponsorship effect. The role of the DLPFC in cognitive control and emotion regulation suggests that it removes the influence of a monetary favor by controlling responses in known valuation regions of the brain including the the VMPFC. PMID- 21646527 TI - Tubular network formation protects mitochondria from autophagosomal degradation during nutrient starvation. AB - Mitochondria are highly dynamic organelles that mediate essential cell functions such as apoptosis and cell-cycle control in addition to their role as efficient ATP generators. Mitochondrial morphology changes are tightly regulated, and their shape can shift between small, fragmented units and larger networks of elongated mitochondria. We demonstrate that mitochondrial elements become significantly elongated and interconnected shortly after nutrient depletion. This mitochondrial morphological shift depends on the type of starvation, with an additive effect observed when multiple nutrients are depleted simultaneously. We further show that starvation-induced mitochondrial elongation is mediated by down-regulation of dynamin-related protein 1 (Drp1) through modulation of two Drp1 phosphorylation sites, leading to unopposed mitochondrial fusion. Finally, we establish that mitochondrial tubulation upon nutrient deprivation protects mitochondria from autophagosomal degradation, which could permit mitochondria to maximize energy production and supply autophagosomal membranes during starvation. PMID- 21646528 TI - Morphology, body proportions, and postcranial hypertrophy of a female Neandertal from the Sima de las Palomas, southeastern Spain. AB - Considerations of Neandertal geographical variation have been hampered by the dearth of remains from Mediterranean Europe and the absence there of sufficiently complete associated postcrania. The 2006 and 2007 excavation of an articulated partial skeleton of a small adult female Neandertal at the Sima de las Palomas, Murcia, southeastern Spain (Sima de las Palomas 96) provides substantial and secure information on body proportions among southern European Neandertals, as well as further documenting the nature of Neandertal biology in southern Iberia. The remains exhibit a suite of cranial, mandibular, dental, and postcranial features, of both Neandertals and archaic Homo generally, that distinguish them from contemporary and subsequent early modern humans. Its lower limbs exhibit the robustness of later Pleistocene Homo generally, and its upper limbs conform to the pattern of elevated robustness of the Neandertals. Its body proportions, including relative clavicular length, distal limb segment lengths, and body mass to stature indicators, conform to the "cold-adapted" pattern of more northern Neandertals. Palomas 96 therefore documents the presence of a suite of "Neandertal" characteristics in southern Iberia and, along with its small body size, the more "Arctic" body proportions of other European Neandertals despite the warmer climate of southern Iberia during marine isotope stage 3. PMID- 21646529 TI - Evolution of the mitochondrial fusion-fission cycle and its role in aging. AB - Mitochondria are organelles of eukaryotic cells that contain their own genetic material and evolved from prokaryotic ancestors some 2 billion years ago. They are the main source of the cell's energy supply and are involved in such important processes as apoptosis, mitochondrial diseases, and aging. During recent years it also became apparent that mitochondria display a complex dynamical behavior of fission and fusion, the function of which is as yet unknown. In this paper we develop a concise theory that explains why fusion and fission have evolved, how these processes are related to the accumulation of mitochondrial mutants during aging, why the mitochondrial DNA has to be located close to the respiration complexes where most radicals are generated, and what selection pressures shaped the slightly different structure of animal and plant mitochondria. We believe that this "organelle control" theory will help in understanding key processes involved in the evolution of the mitochondrial genome and the aging process. PMID- 21646531 TI - Jellyfish blooms result in a major microbial respiratory sink of carbon in marine systems. AB - Jellyfish blooms occur in many estuarine and coastal regions and may be increasing in their magnitude and extent worldwide. Voracious jellyfish predation impacts food webs by converting large quantities of carbon (C), fixed by primary producers and consumed by secondary producers, into gelatinous biomass, which restricts C transfer to higher trophic levels because jellyfish are not readily consumed by other predators. In addition, jellyfish release colloidal and dissolved organic matter (jelly-DOM), and could further influence the functioning of coastal systems by altering microbial nutrient and DOM pathways, yet the links between jellyfish and bacterioplankton metabolism and community structure are unknown. Here we report that jellyfish released substantial quantities of extremely labile C-rich DOM, relative to nitrogen (25.6 +/- 31.6 C:1N), which was quickly metabolized by bacterioplankton at uptake rates two to six times that of bulk DOM pools. When jelly-DOM was consumed it was shunted toward bacterial respiration rather than production, significantly reducing bacterial growth efficiencies by 10% to 15%. Jelly-DOM also favored the rapid growth and dominance of specific bacterial phylogenetic groups (primarily gamma-proteobacteria) that were rare in ambient waters, implying that jelly-DOM was channeled through a small component of the in situ microbial assemblage and thus induced large changes in community composition. Our findings suggest major shifts in microbial structure and function associated with jellyfish blooms, and a large detour of C toward bacterial CO(2) production and away from higher trophic levels. These results further suggest fundamental transformations in the biogeochemical functioning and biological structure of food webs associated with jellyfish blooms. PMID- 21646532 TI - Aphids alter host-plant nitrogen isotope fractionation. AB - Plant sap-feeding insects and blood-feeding parasites are frequently depleted in (15)N relative to their diet. Unfortunately, most fluid-feeder/host nitrogen stable-isotope studies simply report stable-isotope signatures, but few attempt to elucidate the mechanism of isotopic trophic depletion. Here we address this deficit by investigating the nitrogen stable-isotope dynamics of a fluid-feeding herbivore-host plant system: the green peach aphid, Myzus persicae, feeding on multiple brassicaceous host plants. M. persicae was consistently more than 60/00 depleted in (15)N relative to their hosts, although aphid colonized plants were 1.50/00 to 2.00/00 enriched in (15)N relative to uncolonized control plants. Isotopic depletion of aphids relative to hosts was strongly related to host nitrogen content. We tested whether the concomitant aphid (15)N depletion and host (15)N enrichment was coupled by isotopic mass balance and determined that aphid (15)N depletion and host (15)N enrichment are uncoupled processes. We hypothesized that colonized plants would have higher nitrate reductase activity than uncolonized plants because previous studies had demonstrated that high nitrate reductase activity under substrate-limiting conditions can result in increased plant delta(15)N values. Consistent with our hypothesis, nitrate reductase activity in colonized plants was twice that of uncolonized plants. This study offers two important insights that are likely applicable to understanding nitrogen dynamics in fluid-feeder/host systems. First, isotopic separation of aphid and host depends on nitrogen availability. Second, aphid colonization alters host nitrogen metabolism and subsequently host nitrogen stable-isotope signature. Notably, this work establishes a metabolic framework for future hypothesis-driven studies focused on aphid manipulation of host nitrogen metabolism. PMID- 21646530 TI - Artificial selection for a green revolution gene during japonica rice domestication. AB - The semidwarf phenotype has been extensively selected during modern crop breeding as an agronomically important trait. Introduction of the semidwarf gene, semi dwarf1 (sd1), which encodes a gibberellin biosynthesis enzyme, made significant contributions to the "green revolution" in rice (Oryza sativa L.). Here we report that SD1 was involved not only in modern breeding including the green revolution, but also in early steps of rice domestication. We identified two SNPs in O. sativa subspecies (ssp.) japonica SD1 as functional nucleotide polymorphisms (FNPs) responsible for shorter culm length and low gibberellin biosynthetic activity. Genetic diversity analysis among O. sativa ssp. japonica and indica, along with their wild ancestor O. rufipogon Griff, revealed that these FNPs clearly differentiate the japonica landrace and O. rufipogon. We also found a dramatic reduction in nucleotide diversity around SD1 only in the japonica landrace, not in the indica landrace or O. rufipogon. These findings indicate that SD1 has been subjected to artificial selection in rice evolution and that the FNPs participated in japonica domestication, suggesting that ancient humans already used the green revolution gene. PMID- 21646534 TI - Improving model fidelity and sensitivity for complex systems through empirical information theory. AB - In many situations in contemporary science and engineering, the analysis and prediction of crucial phenomena occur often through complex dynamical equations that have significant model errors compared with the true signal in nature. Here, a systematic information theoretic framework is developed to improve model fidelity and sensitivity for complex systems including perturbation formulas and multimodel ensembles that can be utilized to improve both aspects of model error simultaneously. A suite of unambiguous test models is utilized to demonstrate facets of the proposed framework. These results include simple examples of imperfect models with perfect equilibrium statistical fidelity where there are intrinsic natural barriers to improving imperfect model sensitivity. Linear stochastic models with multiple spatiotemporal scales are utilized to demonstrate this information theoretic approach to equilibrium sensitivity, the role of increasing spatial resolution in the information metric for model error, and the ability of imperfect models to capture the true sensitivity. Finally, an instructive statistically nonlinear model with many degrees of freedom, mimicking the observed non-Gaussian statistical behavior of tracers in the atmosphere, with corresponding imperfect eddy-diffusivity parameterization models are utilized here. They demonstrate the important role of additional stochastic forcing of imperfect models in order to systematically improve the information theoretic measures of fidelity and sensitivity developed here. PMID- 21646533 TI - Mimivirus shows dramatic genome reduction after intraamoebal culture. AB - Most phagocytic protist viruses have large particles and genomes as well as many laterally acquired genes that may be associated with a sympatric intracellular life (a community-associated lifestyle with viruses, bacteria, and eukaryotes) and the presence of virophages. By subculturing Mimivirus 150 times in a germ free amoebal host, we observed the emergence of a bald form of the virus that lacked surface fibers and replicated in a morphologically different type of viral factory. When studying a 0.40-MUm filtered cloned particle, we found that its genome size shifted from 1.2 (M1) to 0.993 Mb (M4), mainly due to large deletions occurring at both ends of the genome. Some of the lost genes are encoding enzymes required for posttranslational modification of the structural viral proteins, such as glycosyltransferases and ankyrin repeat proteins. Proteomic analysis allowed identification of three proteins, probably required for the assembly of virus fibers. The genes for two of these were found to be deleted from the M4 virus genome. The proteins associated with fibers are highly antigenic and can be recognized by mouse and human antimimivirus antibodies. In addition, the bald strain (M4) was not able to propagate the sputnik virophage. Overall, the Mimivirus transition from a sympatric to an allopatric lifestyle was associated with a stepwise genome reduction and the production of a predominantly bald virophage resistant strain. The new axenic ecosystem allowed the allopatric Mimivirus to lose unnecessary genes that might be involved in the control of competitors. PMID- 21646535 TI - Isolation of an activator-dependent, promoter-specific chromatin remodeling factor. AB - Repressed PHO5 gene chromatin, isolated from yeast in the native state, was remodeled by yeast extract in a gene activator-dependent, ATP-dependent manner. The product of the reaction bore the hallmark of the process in vivo, the selective removal of promoter nucleosomes, without effect on open reading frame nucleosomes. Fractionation of the extract identified a single protein, chromodomain helicase DNA binding protein 1 (Chd1), capable of the remodeling activity. Deletion of the CHD1 gene in an isw1Delta pho80Delta strain abolished PHO5 gene expression, demonstrating the relevance of the remodeling reaction in vitro to the process in vivo. PMID- 21646536 TI - Light-induced water oxidation at silicon electrodes functionalized with a cobalt oxygen-evolving catalyst. AB - Integrating a silicon solar cell with a recently developed cobalt-based water splitting catalyst (Co-Pi) yields a robust, monolithic, photo-assisted anode for the solar fuels process of water splitting to O(2) at neutral pH. Deposition of the Co-Pi catalyst on the Indium Tin Oxide (ITO)-passivated p-side of a np-Si junction enables the majority of the voltage generated by the solar cell to be utilized for driving the water-splitting reaction. Operation under neutral pH conditions fosters enhanced stability of the anode as compared to operation under alkaline conditions (pH 14) for which long-term stability is much more problematic. This demonstration of a simple, robust construct for photo-assisted water splitting is an important step towards the development of inexpensive direct solar-to-fuel energy conversion technologies. PMID- 21646537 TI - Complete reconstruction of an enzyme-inhibitor binding process by molecular dynamics simulations. AB - The understanding of protein-ligand binding is of critical importance for biomedical research, yet the process itself has been very difficult to study because of its intrinsically dynamic character. Here, we have been able to quantitatively reconstruct the complete binding process of the enzyme-inhibitor complex trypsin-benzamidine by performing 495 molecular dynamics simulations of free ligand binding of 100 ns each, 187 of which produced binding events with an rmsd less than 2 A compared to the crystal structure. The binding paths obtained are able to capture the kinetic pathway of the inhibitor diffusing from solvent (S0) to the bound (S4) state passing through two metastable intermediate states S2 and S3. Rather than directly entering the binding pocket the inhibitor appears to roll on the surface of the protein in its transition between S3 and the final binding pocket, whereas the transition between S2 and the bound pose requires rediffusion to S3. An estimation of the standard free energy of binding gives DeltaG degrees = -5.2 +/- 0.4 kcal/mol (cf. the experimental value -6.2 kcal/mol), and a two-states kinetic model k(on) = (1.5 +/- 0.2) * 10(8) M(-1) s( 1) and k(off) = (9.5 +/- 3.3) * 10(4) s(-1) for unbound to bound transitions. The ability to reconstruct by simple diffusion the binding pathway of an enzyme inhibitor binding process demonstrates the predictive power of unconventional high-throughput molecular simulations. Moreover, the methodology is directly applicable to other molecular systems and thus of general interest in biomedical and pharmaceutical research. PMID- 21646538 TI - The Era GTPase recognizes the GAUCACCUCC sequence and binds helix 45 near the 3' end of 16S rRNA. AB - Era, composed of a GTPase domain and a K homology domain, is essential for bacterial cell viability. It is required for the maturation of 16S rRNA and assembly of the 30S ribosomal subunit. We showed previously that the protein recognizes nine nucleotides ( ) near the 3' end of 16S rRNA, and that this recognition stimulates GTP-hydrolyzing activity of Era. In all three kingdoms of life, the sequence and helix 45 (h45) (nucleotides 1506-1529) are highly conserved. It has been shown that the to double mutation severely affects the viability of bacteria. However, whether Era interacts with G1530 and/or h45 and whether such interactions (if any) contribute to the stimulation of Era's GTPase activity were not known. Here, we report two RNA structures that contain nucleotides 1506-1542 (RNA301), one in complex with Era and GDPNP (GNP), a nonhydrolysable GTP-analogue, and the other in complex with Era, GNP, and the KsgA methyltransferase. The structures show that Era recognizes 10 nucleotides, including G1530, and that Era also binds h45. Moreover, GTPase assay experiments show that G1530 does not stimulate Era's GTPase activity. Rather, A1531 and A1534 are most important for stimulation and h45 further contributes to the stimulation. Although G1530 does not contribute to the intrinsic GTPase activity of Era, its interaction with Era is important for binding and is essential for the protein to function, leading to the discovery of a new cold-sensitive phenotype of Era. PMID- 21646539 TI - Interference by clustered regularly interspaced short palindromic repeat (CRISPR) RNA is governed by a seed sequence. AB - Prokaryotic clustered regularly interspaced short palindromic repeat (CRISPR)/Cas (CRISPR-associated sequences) systems provide adaptive immunity against viruses when a spacer sequence of small CRISPR RNA (crRNA) matches a protospacer sequence in the viral genome. Viruses that escape CRISPR/Cas resistance carry point mutations in protospacers, though not all protospacer mutations lead to escape. Here, we show that in the case of Escherichia coli subtype CRISPR/Cas system, the requirements for crRNA matching are strict only for a seven-nucleotide seed region of a protospacer immediately following the essential protospacer-adjacent motif. Mutations in the seed region abolish CRISPR/Cas mediated immunity by reducing the binding affinity of the crRNA-guided Cascade complex to protospacer DNA. We propose that the crRNA seed sequence plays a role in the initial scanning of invader DNA for a match, before base pairing of the full-length spacer occurs, which may enhance the protospacer locating efficiency of the E. coli Cascade complex. In agreement with this proposal, single or multiple mutations within the protospacer but outside the seed region do not lead to escape. The relaxed specificity of the CRISPR/Cas system limits escape possibilities and allows a single crRNA to effectively target numerous related viruses. PMID- 21646540 TI - 2D NMR-spectroscopic screening reveals polyketides in ladybugs. AB - Small molecules of biological origin continue to yield the most promising leads for drug design, but systematic approaches for exploring nature's cache of structural diversity are lacking. Here, we demonstrate the use of 2D NMR spectroscopy to screen a library of biorationally selected insect metabolite samples for partial structures indicating the presence of new chemical entities. This NMR-spectroscopic survey enabled detection of novel compounds in complex metabolite mixtures without prior fractionation or isolation. Our screen led to discovery and subsequent isolation of two families of tricyclic pyrones in Delphastus catalinae, a tiny ladybird beetle that is employed commercially as a biological pest control agent. The D. catalinae pyrones are based on 23-carbon polyketide chains forming 1,11-dioxo-2,6,10-trioxaanthracene and 4,8-dioxo-1,9,13 trioxaanthracene derivatives, representing ring systems not previously found in nature. This study highlights the utility of 2D NMR-spectroscopic screening for exploring nature's structure space and suggests that insect metabolomes remain vastly underexplored. PMID- 21646541 TI - Loss of the miR-21 allele elevates the expression of its target genes and reduces tumorigenesis. AB - MicroRNA 21 (miR-21) is overexpressed in virtually all types of carcinomas and various types of hematological malignancies. To determine whether miR-21 promotes tumor development in vivo, we knocked out the miR-21 allele in mice. In response to the 7,12-dimethylbenz[a]anthracene (DMBA)/12-O-tetradecanoylphorbol-13-acetate mouse skin carcinogenesis protocol, miR-21-null mice showed a significant reduction in papilloma formation compared with wild-type mice. We revealed that cellular apoptosis was elevated and cell proliferation was decreased in mice deficient of miR-21 compared to wild-type animals. In addition, we found that a large number of validated or predicted miR-21 target genes were up-regulated in miR-21-null keratinocytes, which are precursor cells to skin papillomas. Specifically, up-regulation of Spry1, Pten, and Pdcd4 when miR-21 was ablated coincided with reduced phosphorylation of ERK, AKT, and JNK, three major downstream effectors of Ras activation that plays a predominant role in DMBA initiated skin carcinogenesis. These results provide in vivo evidence that miR-21 exerts its oncogenic function through negatively regulating its target genes. PMID- 21646542 TI - Hund's rule in superatoms with transition metal impurities. AB - The quantum states in metal clusters bunch into supershells with associated orbitals having shapes resembling those in atoms, giving rise to the concept that selected clusters could mimic the characteristics of atoms and be classified as superatoms. Unlike atoms, the superatom orbitals span over multiple atoms and the filling of orbitals does not usually exhibit Hund's rule seen in atoms. Here, we demonstrate the possibility of enhancing exchange splitting in superatom shells via a composite cluster of a central transition metal and surrounding nearly free electron metal atoms. The transition metal d states hybridize with superatom D states and result in enhanced splitting between the majority and minority sets where the moment and the splitting can be controlled by the nature of the central atom. We demonstrate these findings through studies on TMMg(n) clusters where TM is a 3d atom. The clusters exhibit Hund's filling, opening the pathway to superatoms with magnetic shells. PMID- 21646543 TI - Realization of a Knill-Laflamme-Milburn controlled-NOT photonic quantum circuit combining effective optical nonlinearities. AB - Quantum information science addresses how uniquely quantum mechanical phenomena such as superposition and entanglement can enhance communication, information processing, and precision measurement. Photons are appealing for their low-noise, light-speed transmission and ease of manipulation using conventional optical components. However, the lack of highly efficient optical Kerr nonlinearities at the single photon level was a major obstacle. In a breakthrough, Knill, Laflamme, and Milburn (KLM) showed that such an efficient nonlinearity can be achieved using only linear optical elements, auxiliary photons, and measurement [Knill E, Laflamme R, Milburn GJ (2001) Nature 409:46-52]. KLM proposed a heralded controlled-NOT (CNOT) gate for scalable quantum computation using a photonic quantum circuit to combine two such nonlinear elements. Here we experimentally demonstrate a KLM CNOT gate. We developed a stable architecture to realize the required four-photon network of nested multiple interferometers based on a displaced-Sagnac interferometer and several partially polarizing beamsplitters. This result confirms the first step in the original KLM "recipe" for all-optical quantum computation, and should be useful for on-demand entanglement generation and purification. Optical quantum circuits combining giant optical nonlinearities may find wide applications in quantum information processing, communication, and sensing. PMID- 21646544 TI - Hyperinsulinemia leads to uncoupled insulin regulation of the GLUT4 glucose transporter and the FoxO1 transcription factor. AB - Insulin resistance is a component of the metabolic syndrome and Type 2 diabetes. It has been recently shown that in liver insulin resistance is not complete. This so-called selective insulin resistance is characterized by defective insulin inhibition of hepatic glucose output while insulin-induced lipogenesis is maintained. How this occurs and whether uncoupled insulin action develops in other tissues is unknown. Here we show in a model of chronic hyperinsulinemia that adipocytes develop selective insulin resistance in which translocation of the GLUT4 glucose transporter to the cell surface is blunted yet nuclear exclusion of the FoxO1 transcription factor is preserved, rendering uncoupled insulin-controlled carbohydrate and lipid metabolisms. We found that in adipocytes FoxO1 nuclear exclusion has a lower half-maximal insulin dose than GLUT4 translocation, and it is because of this inherent greater sensitivity that control of FoxO1 by physiological insulin concentrations is maintained in adipocytes with compromised insulin signaling. Pharmacological and genetic interventions revealed that insulin regulates GLUT4 and FoxO1 through the PI3 kinase isoform p110alpha, although FoxO1 showed higher sensitivity to p110alpha activity than GLUT4. Transient down-regulation and overexpression of Akt isoforms in adipocytes demonstrated that insulin-activated PI3-kinase signals to GLUT4 primarily through Akt2 kinase, whereas Akt1 and Akt2 signal to FoxO1. We propose that the lower threshold of insulin activity for FoxO1's nuclear exclusion is in part due to its regulation by both Akt isoforms. Identification of uncoupled insulin action in adipocytes suggests this condition might be a general phenomenon of insulin target tissues contributing to insulin resistance's pathophysiology. PMID- 21646545 TI - Assembly mechanism is the key determinant of the dosage sensitivity of a phage structural protein. AB - Altering the expression level of proteins that are subunits of complexes has been proposed to be particularly detrimental because the resulting stoichiometric imbalance among components would lead to misassembly of the complex. Here we show that assembly of the phage HK97 connector complex is severely inhibited by the overexpression of one of its component proteins, gp6. However, this effect is a result of the unusual mechanism by which the oligomerization and assembly of gp6 are controlled. Alteration of this mechanism by single amino acid substitutions leads to a reversal of the response to gp6 overexpression. Surprisingly, the binding partner of gp6 within the phage particle is expressed at a 500-fold higher concentration despite their identical stoichiometry within the complex. Our data emphasize that a generalized prediction of the effects of changes in the expression level of protein complex subunits is very difficult because these effects are dependent upon assembly mechanism. PMID- 21646546 TI - Crystal structure of the MID-PIWI lobe of a eukaryotic Argonaute protein. AB - Argonaute proteins (AGOs) are essential effectors in RNA-mediated gene silencing pathways. They are characterized by a bilobal architecture, in which one lobe contains the N-terminal and PAZ domains and the other contains the MID and PIWI domains. Here, we present the first crystal structure of the MID-PIWI lobe from a eukaryotic AGO, the Neurospora crassa QDE-2 protein. Compared to prokaryotic AGOs, the domain orientation is conserved, indicating a conserved mode of nucleic acid binding. The PIWI domain shows an adaptable surface loop next to a eukaryote specific alpha-helical insertion, which are both likely to contact the PAZ domain in a conformation-dependent manner to sense the functional state of the protein. The MID-PIWI interface is hydrophilic and buries residues that were previously thought to participate directly in the allosteric regulation of guide RNA binding. The interface includes the binding pocket for the guide RNA 5' end, and residues from both domains contribute to binding. Accordingly, micro-RNA (miRNA) binding is particularly sensitive to alteration in the MID-PIWI interface in Drosophila melanogaster AGO1 in vivo. The structure of the QDE-2 MID-PIWI lobe provides molecular and mechanistic insight into eukaryotic AGOs and has significant implications for understanding the role of these proteins in silencing. PMID- 21646547 TI - Levels of p27 sensitize to dual PI3K/mTOR inhibition. AB - Constitutive activation of the phosphatidylinositol 3-kinase (PI3K)/AKT/mTOR signaling cascade occurs in a variety of human malignancies, where it sustains tumor cell proliferation and survival. Pharmacologic blockade of this pathway exerts antineoplastic activity by triggering apoptosis and/or cell-cycle arrest. Pituitary adenomas show activation of the PI3K/AKT/mTOR pathway, but only a fraction of them respond in vitro to the antiproliferative action of rapamycin and RAD001 (mTOR inhibitors), possibly because of the described negative feedback loop on AKT which reactivates the signaling cascade. Rats affected by the multiple endocrine neoplasia-like syndrome (MENX) develop pituitary adenomas showing increased activated AKT. In this study, we comparatively investigated the antitumor potential of the novel dual PI3K/mTOR inhibitor NVP-BEZ235 and the single mTOR inhibitor RAD001 on rat pituitary adenoma cells in primary culture. NVP-BEZ235 inhibits the PI3K pathway both upstream and downstream of AKT, thereby preventing the negative feedback loop. NVP-BEZ235 was more effective than RAD001 in reducing cell viability of pituitary adenomas. Consistently, NVP-BEZ235 treatment decreased Akt and S6 phosphorylation and triggered apoptosis. Because MENX is caused by a germline loss-of-function mutation in the cell-cycle inhibitor p27Kip1, we investigated the relationship between this defect and response to NVP-BEZ235 treatment. The levels of p27Kip1 positively correlate with the response to NVP-BEZ235 treatment. Combined treatment with NVP-BEZ235 and the proteasome inhibitor bortezomib, which increases p27Kip1 amount, shows synergistic antiproliferative effects on pituitary adenoma cells. Our data suggest that NVP-BEZ235 may represent an effective therapeutic modality for pituitary adenomas and that p27Kip1 levels represent a potential predictor of response to dual PI3K/mTOR inhibition. PMID- 21646548 TI - Infant distress: moving toward concept clarity. AB - Infants' inability to articulate their pain and distress and the diverse range of behavioral responses evoked by painful stimuli may partially explain the challenges associated with the treatment of pain in neonates and infants. To assist nurses in distinguishing nonspecific distress responses from specific pain responses, a concept analysis using Wilson's method was preformed. An evaluation of published, peer reviewed literature referencing distress in neonates and infants was conducted to extract meaningful information related to distress. This article examines the essential features, antecedents, and consequences of infant distress to generate an evidence-based definition that has relevance for neonatal and pediatric research and clinical practice. PMID- 21646549 TI - Stereotype threat and female communication styles. AB - A large body of research has documented the performance-debilitating effects of stereotype threat for individuals, but there is a paucity of research exploring interpersonal consequences of stereotype threat. Two experiments tested the hypothesis that stereotype threat would change the style in which women communicate. Results indicate that women who experience stereotype threat regarding leadership abilities react against the stereotype by adopting a more masculine communication style. Study 2 provides evidence that self-affirmation eliminates this effect of stereotype threat on women's communication styles. A third study demonstrates an ironic consequence of this effect of stereotype threat on women's communication--when women under stereotype threat adopt a more masculine communication style, they are rated as less warm and likeable, and evaluators indicate less willingness to comply with their requests. Theoretical and practical implications of these findings are discussed. PMID- 21646550 TI - Comparative evaluation of radiation treatments for clinically localized prostate cancer: an updated systematic review. AB - BACKGROUND: Radiation therapy is one of many treatment options for patients with prostate cancer. PURPOSE: To update findings on the clinical and biochemical outcomes of radiation therapies for localized prostate cancer. DATA SOURCES: MEDLINE (2007 through March 2011) and the Cochrane Central Register of Controlled Trials (2007 through March 2011). STUDY SELECTION: Published English-language comparative studies involving adults with localized prostate cancer who either had first-line radiation therapy or received no initial treatment. DATA EXTRACTION: 6 researchers extracted information on study design, potential bias, sample characteristics, interventions, and outcomes and rated the strength of overall evidence. Data for each study were extracted by 1 reviewer and confirmed by another. DATA SYNTHESIS: 75 studies (10 randomized, controlled trials [RCTs] and 65 nonrandomized studies) met the inclusion criteria. No RCTs compared radiation therapy with no treatment or no initial treatment. Among the 10 RCTs, 2 compared combinations of radiation therapies, 7 compared doses and fraction sizes of external-beam radiation therapy (EBRT), and 1 compared forms of low-dose rate radiation therapy. Heterogeneous outcomes were analyzed. Overall, moderate strength evidence consistently showed that a higher EBRT dose was associated with increased rates of long-term biochemical control compared with lower EBRT dose. The body of evidence was rated as insufficient for all other comparisons. LIMITATIONS: Studies inconsistently defined and reported outcomes. Much of the available evidence comes from observational studies with treatment selection biases. CONCLUSION: A lack of high-quality comparative evidence precludes conclusions about the efficacy of radiation treatments compared with no treatments for localized prostate cancer. PRIMARY FUNDING SOURCE: Agency for Healthcare Research and Quality. PMID- 21646551 TI - Effects of electronic health information technology implementation on nursing home resident outcomes. AB - OBJECTIVE: To examine the effects of electronic health information technology (HIT) on nursing home residents. METHODS: The study evaluated the impact of implementing a comprehensive HIT system on resident clinical, functional, and quality of care outcome indicators as well as measures of resident awareness of and satisfaction with the technology. The study used a prospective, quasi experimental design, directly assessing 761 nursing home residents in 10 urban and suburban nursing homes in the greater New York City area. RESULTS: No statistically significant impact of the introduction of HIT on residents was found on any outcomes, with the exception of a significant negative effect on behavioral symptoms. Residents' subjective assessment of the HIT intervention were generally positive. DISCUSSION: The absence of effects on most indicators is encouraging for the future development of HIT in nursing homes. The single negative finding suggests that further investigation is needed on possible impact on resident behavior. PMID- 21646552 TI - Summaries for patients: smoking and peripheral artery disease in women. PMID- 21646553 TI - In the clinic. Delirium. AB - This issue provides a clinical overview of delirium focusing on prevention, diagnosis, treatment, practice improvement, and patient information. Readers can complete the accompanying CME quiz for 1.5 credits. Only ACP members and individual subscribers can access the electronic features of In the Clinic. Non subscribers who wish to access this issue of In the Clinic can elect "Pay for View." Subscribers can receive 1.5 category 1 CME credits by completing the CME quiz that accompanies this issue of In the Clinic. The content of In the Clinic is drawn from the clinical information and education resources of the American College of Physicians (ACP), including PIER (Physicians' Information and Education Resource) and MKSAP (Medical Knowledge and Self Assessment Program). Annals of Internal Medicine editors develop In the Clinic from these primary sources in collaboration with the ACP's Medical Education and Publishing division and with assistance of science writers and physician writers. Editorial consultants from PIER and MKSAP provide expert review of the content. Readers who are interested in these primary resources for more detail can consult www.acponline.org, http://pier.acponline.org, and other resources referenced within each issue of In the Clinic. PMID- 21646554 TI - Performance of 4 clinical decision rules in the diagnostic management of acute pulmonary embolism: a prospective cohort study. AB - BACKGROUND: Several clinical decision rules (CDRs) are available to exclude acute pulmonary embolism (PE), but they have not been directly compared. OBJECTIVE: To directly compare the performance of 4 CDRs (Wells rule, revised Geneva score, simplified Wells rule, and simplified revised Geneva score) in combination with d dimer testing to exclude PE. DESIGN: Prospective cohort study. SETTING: 7 hospitals in the Netherlands. PATIENTS: 807 consecutive patients with suspected acute PE. INTERVENTION: The clinical probability of PE was assessed by using a computer program that calculated all CDRs and indicated the next diagnostic step. Results of the CDRs and d-dimer tests guided clinical care. MEASUREMENTS: Results of the CDRs were compared with the prevalence of PE identified by computed tomography or venous thromboembolism at 3-month follow-up. RESULTS: Prevalence of PE was 23%. The proportion of patients categorized as PE-unlikely ranged from 62% (simplified Wells rule) to 72% (Wells rule). Combined with a normal d-dimer result, the CDRs excluded PE in 22% to 24% of patients. The total failure rates of the CDR and d-dimer combinations were similar (1 failure, 0.5% to 0.6% [upper limit 95% CI, 2.9% to 3.1%]). Even though 30% of patients had discordant CDR outcomes, PE was not detected in any patient with discordant CDRs and a normal d dimer result. LIMITATION: Management was based on a combination of decision rules and d-dimer testing rather than only 1 CDR combined with d-dimer testing. CONCLUSION: All 4 CDRs show similar performance for exclusion of acute PE in combination with a normal d-dimer result. This prospective validation indicates that the simplified scores may be used in clinical practice. PRIMARY FUNDING SOURCE: Academic Medical Center, VU University Medical Center, Rijnstate Hospital, Leiden University Medical Center, Maastricht University Medical Center, Erasmus Medical Center, and Maasstad Hospital. PMID- 21646555 TI - Smoking, smoking cessation, [corrected] and risk for symptomatic peripheral artery disease in women: a cohort study. AB - BACKGROUND: Smoking has a well-documented detrimental effect on risk for myocardial infarction and stroke, but less information is available regarding peripheral artery disease (PAD), particularly among women. OBJECTIVE: To prospectively assess the association of current smoking status, cumulative smoking exposure, and smoking cessation with incident symptomatic PAD in women. DESIGN: Prospective cohort study. SETTING: U.S. female health care professionals in the Women's Health Study. PARTICIPANTS: 39,825 women with no cardiovascular disease who were prospectively followed for a median of 12.7 years. MEASUREMENTS: Incidence of symptomatic PAD. Cox proportional hazards models were used to compare PAD risk across smoking categories. RESULTS: 178 confirmed PAD events occurred. Across the 4 smoking categories (never, former, <15 cigarettes/d, and >=15 cigarettes/d), age-adjusted incidence rates were 0.12, 0.34, 0.95, and 1.63 per 1000 person-years of follow-up, respectively. Multivariate adjustment had little effect on this relationship (adjusted hazard ratios [HRs], 3.14 [95% CI, 2.01 to 4.90], 8.93 [CI, 5.02 to 15.89], and 16.95 [CI, 10.77 to 26.67], respectively, vs. women who never smoked). Additional adjustment for high sensitivity C-reactive protein and soluble intercellular adhesion molecule-1 levels among women with available blood samples (28,314 participants, 117 events) attenuated risk estimates (HR, 5.58 [CI, 2.61 to 11.93] for smoking <15 cigarettes/d and 9.52 [CI, 5.17 to 17.53] for smoking >=15 cigarettes/d). Lifetime exposure showed a strong dose-response relationship; fully adjusted HRs for smoking abstinence of fewer than 10, 10 to 29, and 30 or more pack-years were 2.52 (CI, 1.49 to 4.25), 6.75 (CI, 4.33 to 10.52), and 11.09 (CI, 6.94 to 17.72), respectively. Compared with current smokers, the adjusted HRs for fewer than 10 years, 10 to 20 years, more than 20 years, or lifelong abstinence were 0.39 (CI, 0.24 to 0.66), 0.28 (CI, 0.17 to 0.46), 0.16 (CI, 0.10 to 0.26), and 0.08 (CI, 0.05 to 0.12), respectively. LIMITATION: The use of symptomatic PAD as the a priori primary end point excludes asymptomatic disease. CONCLUSION: Among initially healthy women, smoking is a potent risk factor for symptomatic PAD and was associated with subclinical inflammation. Smoking cessation substantially reduces risk for PAD, but an increased occurrence of PAD persists even among former smokers who maintain abstinence. PRIMARY FUNDING SOURCE: The National Heart, Lung, and Blood Institute and National Cancer Institute. PMID- 21646556 TI - Quality of care for older patients with cancer in the Veterans Health Administration versus the private sector: a cohort study. AB - BACKGROUND: The Veterans Health Administration (VHA) is the largest integrated health care system in the United States. Studies suggest that the VHA provides better preventive care and care for some chronic illnesses than does the private sector. OBJECTIVE: To assess the quality of cancer care for older patients provided by the VHA versus fee-for-service Medicare. DESIGN: Observational study of patients with cancer that was diagnosed between 2001 and 2004 who were followed through 2005. SETTING: VHA and non-VHA hospitals and office-based practices. PATIENTS: Men older than 65 years with incident colorectal, lung, or prostate cancer; lymphoma; or multiple myeloma. MEASUREMENTS: Rates of processes of care for colorectal, lung, or prostate cancer; lymphoma; or multiple myeloma. Rates were adjusted by using propensity score weighting. RESULTS: Compared with the fee-for-service Medicare population, the VHA population received diagnoses of colon (P < 0.001) and rectal (P = 0.007) cancer at earlier stages and had higher adjusted rates of curative surgery for colon cancer (92.7% vs. 90.5%; P < 0.010), standard chemotherapy for diffuse large B-cell non-Hodgkin lymphoma (71.1% vs. 59.3%; P < 0.001), and bisphosphonate therapy for multiple myeloma (62.1% vs. 50.4%; P < 0.001). The VHA population had lower adjusted rates of 3-dimensional conformal or intensity-modulated radiation therapy for prostate cancer treated with external-beam radiation therapy (61.6% vs. 86.0%; P < 0.001). Adjusted rates were similar for 9 other measures. Sensitivity analyses suggest that if patients with cancer in the VHA system have more severe comorbid illness than other patients, rates for most indicators would be higher in the VHA population than in the fee-for-service Medicare population. LIMITATION: This study included only older men and did not include information about performance status, severity of comorbid illness, or patient preferences. CONCLUSION: Care for older men with cancer in the VHA system was generally similar to or better than care for fee-for service Medicare beneficiaries, although adoption of some expensive new technologies may be delayed in the VHA system. PRIMARY FUNDING SOURCE: Department of Veterans Affairs. PMID- 21646557 TI - Synopsis of the National Institute for Health and Clinical Excellence guideline for prevention of delirium. AB - DESCRIPTION: Delirium is common, is often underrecognized, and is associated with poor outcomes and high costs. In July 2010, the National Institute for Health and Clinical Excellence released a guideline that addressed diagnosis, prevention, and management of delirium. This synopsis focuses on the main recommendations about prevention of delirium. METHODS: The National Clinical Guideline Centre developed these guidelines by using standard methodology of the National Institute for Health and Clinical Excellence. A multidisciplinary guideline development group posed review questions, discussed evidence, and formulated the recommendations. To underpin the guideline, a technical team from the National Clinical Guideline Centre systematically reviewed and graded pertinent evidence identified from literature searches of studies published in English to August 2009 and performed health economic modeling. Stakeholder and public comment informed guideline development and modifications. RECOMMENDATIONS: Considering prevention a feasible and cost-effective health strategy, the guideline development group made 13 specific recommendations that addressed the stability of the care environment (both the care team and location) and the provision of a multicomponent intervention package tailored for persons at risk for delirium. The multicomponent intervention package included assessment and modification of key clinical factors that may precipitate delirium, including cognitive impairment or disorientation, dehydration or constipation, hypoxia, infection, immobility or limited mobility, several medications, pain, poor nutrition, sensory impairment, and sleep disturbance. PMID- 21646558 TI - A summary of the methods that the National Clinical Guideline Centre uses to produce clinical guidelines for the National Institute for Health and Clinical Excellence. AB - The National Clinical Guideline Centre develops evidence-based clinical guidelines on behalf of the National Institute for Health and Clinical Excellence in the United Kingdom. These guidelines are developed for the National Health Service in England, Wales, and Northern Ireland and establish recommendations on best practice. The authors summarize the main methods used in development, how evidence from systematic reviews is interpreted to form recommendations, who is involved in the process, and the main outputs that are published. PMID- 21646559 TI - National Clinical Guideline Centre cost-effectiveness assessment for the National Institute for Health and Clinical Excellence. AB - The National Clinical Guideline Centre (NCGC) develops evidence-based clinical guidelines on behalf of the National Institute for Health and Clinical Excellence (NICE) in the United Kingdom. The U.K. Department of Health has commissioned NICE to make recommendations on the basis of both clinical effectiveness and cost effectiveness. This article describes how cost-effectiveness is evaluated and accounted for in NCGC guidelines. Six recent case studies are presented, in which consideration of cost-effectiveness has informed recommendations in various ways for clinical guidelines on alcohol use disorders, chronic obstructive pulmonary disease, glaucoma, lower urinary tract symptoms, non-ST-segment elevation myocardial infarction and unstable angina, and venous thromboembolism prophylaxis. Some of the challenges faced in trying to account for cost effectiveness in clinical guidelines are outlined, as well as some of the difficulties in adapting cost-effectiveness guidelines for other settings. PMID- 21646560 TI - Reconsidering the Veterans Health Administration: a model and a moment for publicly funded health care delivery. PMID- 21646561 TI - Trustworthy clinical guidelines. PMID- 21646562 TI - Trained to avoid primary care. PMID- 21646563 TI - Rates of kidney transplantation in nations with presumed consent. PMID- 21646564 TI - The patient-centered medical home neighbor. PMID- 21646565 TI - Residency training at a crossroads. PMID- 21646566 TI - Residency training at a crossroads. PMID- 21646567 TI - Declining autopsy rates and suicide misclassification: a cross-national analysis of 35 countries. AB - CONTEXT: Suicides are prone to misclassification during death ascertainment procedures. This problem has generated frequent criticism of the validity of suicide mortality statistics. OBJECTIVE: To employ an external measure of the validity of cause-of-death statistics (ie, national autopsy rates) and to examine potential misclassification of suicide across countries from Europe to Central and Northern Asia. DESIGN: Cross-national analysis. SETTING: Thirty-five countries. PARTICIPANTS: Aggregated mortality data. MAIN OUTCOME MEASURES: Data from 35 countries during the period from 1979 to 2007 were used to analyze the association of suicide rates with autopsy rates and death rates of undetermined and ill-defined causes, respectively. Analyses were cross-sectional and longitudinal. RESULTS: Cross-sectionally, a 1% difference in autopsy rates among nations was associated with a suicide rate difference of 0.49 per 100,000 population. Longitudinally, a 1% decrease in the autopsy rate aligned with a decrease of 0.42 per 100,000 population in the suicide rate. These cross sectional and longitudinal associations were robust after adjustment for unemployment, degree of urbanization, and prevalence of undetermined or ill defined deaths. Associations strengthened when analyses were confined to 19 European Union member countries. CONCLUSION: Autopsy rates may spatially and temporally affect the validity of suicide mortality statistics. Caution should be exercised in comparing international suicide rates and evaluating interventions that target suicide rate reduction. PMID- 21646568 TI - Testing the reliability and validity of DSM-IV-TR and ICSD-2 insomnia diagnoses. Results of a multitrait-multimethod analysis. AB - CONTEXT: Distinctive diagnostic classification schemes for insomnia diagnoses are available, but the optimal insomnia nosology has yet to be determined. OBJECTIVES: To test the reliability and validity of insomnia diagnoses listed in the American Psychiatric Association's DSM-IV-TR and the International Classification of Sleep Disorders, second edition (ICSD-2). DESIGN: Multitrait multimethod correlation design. SETTING: Two collaborating university medical centers, with recruitment from January 2004 to February 2009. PARTICIPANTS: A total of 352 adult volunteers (235 of whom were women) who met research diagnostic criteria for insomnia disorder. MAIN OUTCOME MEASURES: Goodness-of-fit ratings of 10 DSM-IV-TR and 37 ICSD-2 insomnia diagnoses for each patient. Ratings were provided by 3 clinician pairs who used distinctive assessment methods to derive diagnostic impressions. Correlations computed within and across clinician pairs were used to test reliability and validity of diagnoses. RESULTS: Findings suggested that the best-supported DSM-IV-TR insomnia categories were insomnia related to another mental disorder, insomnia due to a general medical condition, breathing-related sleep disorder, and circadian rhythm sleep disorder. The category of primary insomnia appeared to have marginal reliability and validity. The best-supported ICSD-2 categories were the insomnias due to a mental disorder and due to a medical condition, obstructive sleep apnea, restless legs syndrome, idiopathic insomnia, and circadian rhythm sleep disorder-delayed sleep phase type. Psychophysiological insomnia and inadequate sleep hygiene received much more variable support across sites, whereas the diagnosis of paradoxical insomnia was poorly supported. CONCLUSIONS: Both the DSM-IV-TR and ICSD-2 provide viable insomnia diagnoses, but findings support selected subtypes from each of the 2 nosologies. Nonetheless, findings regarding the frequently used DSM-IV-TR diagnosis of primary insomnia and its related ICSD-2 subtypes suggest that their poor reliability and validity are perhaps due to significant overlap with comorbid insomnia subtypes. Therefore, alternate diagnostic paradigms should be considered for insomnia classification. PMID- 21646569 TI - Disentangling structural brain alterations associated with violent behavior from those associated with substance use disorders. AB - CONTEXT: Studies aimed at identifying structural brain alterations associated with persistent violent behavior or psychopathy have not adequately accounted for a lifetime history of substance misuse. Thus, alterations in gray matter (GM) volume that have been reported to be correlates of violent behavior and/or psychopathy may instead be related to lifelong substance use disorders (SUDs). OBJECTIVE: To identify alterations in GM volume associated with violent behavior and those associated with lifelong SUDs. DESIGN: Cross-sectional study. SETTING: Participants were recruited from penitentiaries, forensic hospitals, psychiatric outpatient services, and communities in Germany. Structural magnetic resonance imaging was performed at a university hospital. PARTICIPANTS: Four groups of men were compared: 12 men with SUDs who exhibited violent behavior (hereafter referred to as violent offenders), 12 violent offenders without SUDs, 13 men with SUDs who did not exhibit violent behavior (hereafter referred to as nonoffenders), and 14 nonoffenders without SUDs. MAIN OUTCOME MEASURES: Voxel based morphometry was used to analyze high-resolution magnetic resonance imaging scans. Assessments of mental disorders, psychopathy (using the Psychopathy Checklist-Screening Version), aggressive behavior, and impulsivity were conducted by trained clinicians. RESULTS: Compared with nonoffenders, violent offenders presented with a larger GM volume in the amygdala bilaterally, the left nucleus accumbens, and the right caudate head and with less GM volume in the left insula. Men with SUDs exhibited a smaller GM volume in the orbitofrontal cortex, ventromedial prefrontal cortex, and premotor cortex than did men without SUDs. Regression analyses indicated that the alterations in GM volume that distinguished the violent offenders from nonoffenders were associated with psychopathy scores and scores for lifelong aggressive behavior. The GM volumes of the orbitofrontal cortex and prefrontal cortex that distinguished the men with SUDs from the men without SUDs were correlated with scores for response inhibition. CONCLUSIONS: These findings suggest that a greater GM volume in the mesolimbic reward system may be associated with violent behavior and that reduced GM volumes in the prefrontal cortex, orbitofrontal cortex, and premotor area characterize men with SUDs. PMID- 21646570 TI - School bullying perpetration and other childhood risk factors as predictors of adult intimate partner violence perpetration. AB - OBJECTIVE: To assess the relationship between bullying peers as a child and adult intimate partner violence perpetration in a clinic-based sample of adult men. School bullying perpetration and intimate partner violence perpetration are both thought to stem from desire for power and control over others. DESIGN: A cross sectional survey was conducted between January 2005 and December 2006. SETTING: Three urban community health centers in Boston, Massachusetts. PARTICIPANTS: Men aged 18 to 35 years (n = 1491) seeking services at participating community health centers. MAIN EXPOSURE: School bullying perpetration. OUTCOME MEASURE: Past-year physical or sexual violence perpetration against a female partner (intimate partner violence [IPV]). RESULTS: Two-fifths of men reported perpetrating school bullying as a child (n = 610; 40.9%). Men who rarely bullied in school were 1.53 times more likely to perpetrate past-year IPV than men who did not bully (95% confidence interval [CI], 1.02-2.29); this risk was elevated to 3.82 times more likely to perpetrate any past-year IPV for those men who bullied peers frequently (95% CI, 2.55-5.73). CONCLUSIONS: The present study indicates that bullying peers in school as a child, especially frequent bullying perpetration, is associated with increased risk for men's perpetration of IPV as an adult. The effect remains strong after controlling for common prior risk factors for both bullying and IPV perpetration. Future research is needed to discern the mechanisms and underlying root causes of abusive behavior, such as power and control, as a means to prevent violence perpetration across settings and life stages. PMID- 21646571 TI - The introduction of allergenic foods and the development of reported wheezing and eczema in childhood: the Generation R study. AB - OBJECTIVE: To examine whether the timing of introduction of the allergenic foods cow's milk, hen's egg, peanuts, tree nuts, soy, and gluten is associated with eczema and wheezing in children 4 years of age or younger. DESIGN: Population based prospective cohort study from fetal life until young adulthood. SETTING: Rotterdam, the Netherlands, from April 2002 through January 2006. PARTICIPANTS: A total of 6905 preschool children participating in the Generation R study. MAIN EXPOSURE: Timing of introduction of cow's milk, hen's egg, peanuts, tree nuts, soy, and gluten collected by questionnaires at 6 and 12 months of age. MAIN OUTCOME MEASURES: Information on the outcomes eczema and wheezing were obtained by questions from the age-adapted version of the "International Study of Asthma and Allergies in Childhood" core questionnaire and questionnaire data on parentally reported physician diagnosis for eczema. RESULTS: Of 6905 children, wheezing was reported in 31% at age 2 years and in 14% at ages 3 and 4 years. Eczema was reported in 38%, 20%, and 18% of children at the ages of 2, 3, and 4 years, respectively. The introduction of cow's milk, hen's egg, peanuts, tree nuts, soy, and gluten before the age of 6 months was not significantly associated with eczema or wheezing at any age after adjustment for potential confounders (P > .10 for all comparisons). The results did not alter after stratification according to the child's history of cow's milk allergy and parental history of atopy. CONCLUSION: This study does not support the recommendation for delayed introduction of allergenic foods after age 6 months for the prevention of eczema and wheezing. PMID- 21646572 TI - Perceptions about water and increased use of bottled water in minority children. AB - OBJECTIVE: To describe bottled water use and beliefs and attitudes about water among parents of children from different racial/ethnic groups. DESIGN: Cross sectional survey. SETTING: Urban/suburban emergency department. PARTICIPANTS: Parents of children treated between September 2009 and March 2010. MAIN OUTCOME MEASURES: The respondents completed a questionnaire in English or Spanish, describing their use of bottled water and tap water for their children and rating their agreement with a series of belief statements about bottled water and tap water. Logistic regression was used to evaluate the association between bottled water use and beliefs and demographic characteristics with odds ratios (ORs). RESULTS: A total of 632 surveys were completed (35% white, 33% African American, and 32% Latino respondents). African American and Latino parents were more likely to give their children mostly bottled water; minority children were exclusively given bottled water 3 times more often than non-Latino white children (24% vs 8%, P < .01). In logistic regression analysis, the following factors were independently associated with mostly bottled water use: belief that bottled water is safer (OR, 2.4), cleaner (OR, 2.0), better tasting (OR, 2.8), or more convenient (OR, 1.7). After other factors were adjusted for, race/ethnicity, household income, and prior residence outside the United States were not associated with bottled water use. CONCLUSIONS: Minority parents are more likely to exclusively give bottled water to their children. Disparities in bottled water use are driven largely by differences in beliefs and perceptions about water. Interventions to reduce bottled water use among minority families should be based on knowledge of the factors that are related to water use in these communities. PMID- 21646573 TI - Parental and environmental correlates of physical activity of children attending preschool. AB - OBJECTIVE: To determine, using a social-cognitive framework and structural equation modeling, if parent-reported family physical activity (PA) variables are related to PA of young children. DESIGN: Cross-sectional study. SETTING: Children attending 23 preschools in and around Columbia, South Carolina. PARTICIPANTS: Three hundred sixty-nine children (48.0% male and 50.4% black) and their parents. MAIN EXPOSURES: Family variables were reported by parents and included parent PA, parent enjoyment of PA, importance to adults of child playing sports and being active, and family support. MAIN OUTCOME MEASURES: Moderate to vigorous physical activity (MVPA) of children was modeled as a latent variable using PA from direct observation, accelerometers, and parent's perception of the child's athletic coordination. RESULTS: A model of direct and indirect relations of family variables, preschool quality, home PA equipment, and child's enjoyment of PA had acceptable fit (root mean square error of approximation, 0.053; comparative fit index, 0.90). Parent PA, parent enjoyment of PA, and importance of child's PA were significantly related to family support. Family support, quality of preschool attended, home PA equipment, and child's enjoyment of PA were positively related to child's PA. However, there was no direct relationship between parent PA and the child's PA. CONCLUSION: Although parent PA was not directly related to children's MVPA, results showed that parent PA indirectly affects preschool children's MVPA via its influence on family support for children's PA. PMID- 21646574 TI - Pieter Bruegel's Dulle Griet. PMID- 21646575 TI - In vivo imaging of cerebral serotonin transporter and serotonin(2A) receptor binding in 3,4-methylenedioxymethamphetamine (MDMA or "ecstasy") and hallucinogen users. AB - CONTEXT: Both hallucinogens and 3,4-methylenedioxymethamphetamine (MDMA or "ecstasy") have direct agonistic effects on postsynaptic serotonin(2A) receptors, the key site for hallucinogenic actions. In addition, MDMA is a potent releaser and reuptake inhibitor of presynaptic serotonin. OBJECTIVE: To assess the differential effects of MDMA and hallucinogen use on cerebral serotonin transporter (SERT) and serotonin(2A) receptor binding. DESIGN: A positron emission tomography study of 24 young adult drug users and 21 nonusing control participants performed with carbon 11 ((11)C)-labeled 3-amino-4-[2 [(di(methyl)amino)methyl]phenyl]sulfanylbenzonitrile (DASB) and fluorine 18 ((18)F)-labeled altanserin, respectively. Scans were performed in the user group after a minimum drug abstinence period of 11 days, and the group was subdivided into hallucinogen-preferring users (n = 10) and MDMA-preferring users (n = 14). PARTICIPANTS: Twenty-four young adult users of MDMA and/or hallucinogenic drugs and 21 nonusing controls. MAIN OUTCOME MEASURES: In vivo cerebral SERT and serotonin(2A) receptor binding. RESULTS: Compared with nonusers, MDMA-preferring users showed significant decreases in SERT nondisplaceable binding potential (neocortex, -56%; pallidostriatum, -19%; and amygdala, -32%); no significant changes were seen in hallucinogen-preferring users. Both cortical and pallidostriatal SERT nondisplaceable binding potential was negatively correlated with the number of lifetime MDMA exposures, and the time of abstinence from MDMA was positively correlated with subcortical, but not cortical, SERT binding. A small decrease in neocortical serotonin(2A) receptor binding in the serotonin(2A) receptor agonist users (both user groups) was also detected. CONCLUSIONS: We found evidence that MDMA but not hallucinogen use is associated with changes in the cerebral presynaptic serotonergic transmitter system. Because hallucinogenic drugs primarily have serotonin(2A) receptor agonistic actions, we conclude that the negative association between MDMA use and cerebral SERT binding is mediated through a direct presynaptic MDMA effect rather than by the serotonin(2A) agonistic effects of MDMA. Our cross-sectional data suggest that subcortical, but not cortical, recovery of SERT binding might take place after several months of MDMA abstinence. PMID- 21646576 TI - An effectiveness trial of group cognitive behavioral therapy for patients with persistent depressive symptoms in substance abuse treatment. AB - CONTEXT: Although depression frequently co-occurs with substance abuse, few individuals entering substance abuse treatment have access to effective depression treatment. OBJECTIVE: The Building Recovery by Improving Goals, Habits, and Thoughts (BRIGHT) study is a community-based effectiveness trial that compared residential substance abuse treatment with residential treatment plus group cognitive behavioral therapy for depression delivered by substance abuse treatment counselors. We hypothesized that intervention clients would have improved depression and substance use outcomes compared with those of clients receiving usual care. DESIGN: A nonrandomized controlled trial using a quasi experimental intent-to-treat design in which 4 sites were assigned to alternate between the intervention and usual care conditions every 4 months for 21/2 years. SETTING: Four treatment programs in Los Angeles County. PARTICIPANTS: We screened 1262 clients for persistent depressive symptoms (Beck Depression Inventory-II score >17). We assigned 299 clients to receive either usual care (n = 159) or usual care plus the intervention (n = 140). Follow-up rates at 3 and 6 months after the baseline interview were 88.1% and 86.2%, respectively, for usual care and 85.7% and 85.0%, respectively, for the intervention group. INTERVENTION: Sixteen 2-hour group sessions of cognitive behavioral therapy for depression. MAIN OUTCOME MEASURES: Change in depression symptoms, mental health functioning, and days of alcohol and problem substance use. RESULTS: Intervention clients reported significantly fewer depressive symptoms (P < .001 at 3 and 6 months) and had improved mental health functioning (P < .001 at 3 months and P < .01 at 6 months). At 6 months, intervention clients reported fewer drinking days (P < .05) and fewer days of problem substance use (P < .05) on days available. CONCLUSIONS: Providing group cognitive behavioral therapy for depression to clients with persistent depressive symptoms receiving residential substance abuse treatment is associated with improved depression and substance use outcomes. These results provide support for a new model of integrated care. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT01191788. PMID- 21646577 TI - Translational epidemiology in psychiatry: linking population to clinical and basic sciences. AB - Translational research generally refers to the application of knowledge generated by advances in basic sciences research translated into new approaches for diagnosis, prevention, and treatment of disease. This direction is called bench to-bedside. Psychiatry has similarly emphasized the basic sciences as the starting point of translational research. This article introduces the term translational epidemiology for psychiatry research as a bidirectional concept in which the knowledge generated from the bedside or the population can also be translated to the benches of laboratory science. Epidemiologic studies are primarily observational but can generate representative samples, novel designs, and hypotheses that can be translated into more tractable experimental approaches in the clinical and basic sciences. This bedside-to-bench concept has not been explicated in psychiatry, although there are an increasing number of examples in the research literature. This article describes selected epidemiologic designs, providing examples and opportunities for translational research from community surveys and prospective, birth cohort, and family-based designs. Rapid developments in informatics, emphases on large sample collection for genetic and biomarker studies, and interest in personalized medicine--which requires information on relative and absolute risk factors--make this topic timely. The approach described has implications for providing fresh metaphors to communicate complex issues in interdisciplinary collaborations and for training in epidemiology and other sciences in psychiatry. PMID- 21646578 TI - Functional impairment in elderly patients with mild cognitive impairment and mild Alzheimer disease. AB - CONTEXT: The original mild cognitive impairment (MCI) criteria exclude substantial functional deficits, but recent reports suggest otherwise. Identifying the extent, severity, type, and correlates of functional deficits that occur in MCI and mild Alzheimer disease (AD) can aid in early detection of incipient dementia and can identify potential mechanistic pathways to disrupted instrumental activities of daily living (IADLs). OBJECTIVES: To examine the number, type, and severity of functional impairments and to identify the clinical characteristics associated with functional impairment across patients with amnestic MCI (aMCI) and those with mild AD. DESIGN: Study using baseline data from the Alzheimer's Disease Neuroimaging Initiative. SETTING: Multiple research sites in the United States and Canada. Patients Samples included 229 control individuals, 394 patients with aMCI, and 193 patients with AD. MAIN OUTCOME MEASURE: The 10-item Pfeffer Functional Activities Questionnaire (FAQ) assessed function. RESULTS: Informant-reported FAQ deficits were common in patients with aMCI (72.3%) and AD (97.4%) but were rarely self-reported by controls (7.9%). The average severity per FAQ deficit did not differ between patients with aMCI and controls; both were less impaired than patients with AD (P < .001). Two FAQ items (remembering appointments, family occasions, holidays, and medications and assembling tax records, business affairs, or other papers) were specific (specificity estimate, 0.95) in differentiating the control group from the combined aMCI and AD groups (only 34.0% of patients with aMCI and 3.6% of patients with AD had no difficulty with these 2 items). The severity of FAQ deficits in the combined aMCI and AD group was associated with worse Trail Making Test, part A scores and smaller hippocampal volumes (P < .001 for both). Within the aMCI group, functionally intact individuals had greater hippocampal volumes and better Auditory Verbal Learning Test 30-minute delay and Trail Making Test, part A (P < .001 for each) scores compared with individuals with moderate or severe FAQ deficits. Patients with a high number of deficits were more likely to express the apolipoprotein epsilon4 allele (63.8%) compared with patients with no (46.8%) or few (48.4%) functional deficits. CONCLUSIONS: Mild IADL deficits are common in individuals with aMCI and should be incorporated into MCI criteria. Two IADLs--remembering appointments, family occasions, holidays, and medications and assembling tax records, business affairs, or other papers--appear to be characteristic of clinically significant cognitive impairment. In patients with aMCI, impairment in memory and processing speed and greater medial temporal atrophy were associated with greater IADL deficits. PMID- 21646580 TI - A multivariate twin study of obsessive-compulsive symptom dimensions. AB - CONTEXT: Obsessive-compulsive disorder (OCD) is clinically heterogeneous, but it is unclear whether this phenotypic heterogeneity reflects distinct, or partially distinct, etiologic mechanisms. OBJECTIVE: To clarify the structure of the genetic and environmental risk factors for the major symptom dimensions of OCD. DESIGN: Self-report questionnaires and multivariate twin model fitting. SETTING: General community. PARTICIPANTS: A total of 4355 female members of the TwinsUK adult twin register. MAIN OUTCOME MEASURES: Scores on the Obsessive-Compulsive Inventory-Revised and 5 of its subscales (checking, hoarding, obsessing, ordering, and washing). RESULTS: A common pathway model did not fit the data well, indicating that no single latent factor can explain the heterogeneity of OCD. The best-fit multivariate twin model was an independent pathway model, whereby both common and unique genetic and/or environmental factors contribute to the etiology of each symptom dimension. The hoarding dimension had the lowest loading on the common factor and was more influenced by specific genetic effects (54.5% specific). With the exception of hoarding, most of the genetic variance was due to shared genetic factors (ranging from 62.5% to 100%), whereas most of the nonshared environmental variance was due to dimension-specific factors. CONCLUSIONS: Obsessive-compulsive disorder is unlikely to be an etiologically homogeneous condition. There is substantial etiologic overlap across the different OC symptom dimensions, but dimension-specific genetic, and particularly nonshared environmental, factors are at least as important. Hoarding shares the least amount of genetic liability with the remaining symptom dimensions. The results have implications for the current deliberations regarding OCD and the inclusion of a putative hoarding disorder in DSM-5. PMID- 21646579 TI - Collaborative depression care management and disparities in depression treatment and outcomes. AB - CONTEXT: Collaborative depression care management (DCM), by addressing barriers disproportionately affecting patients of racial/ethnic minority and low education, may reduce disparities in depression treatment and outcomes. OBJECTIVE: To examine the effects of DCM on treatment disparities by education and race/ethnicity in older depressed primary care patients. DESIGN: Analysis of data from the randomized controlled trial Prevention of Suicide in Primary Care Elderly: Collaborative Trial (PROSPECT). SETTING: Twenty primary care practices. PARTICIPANTS: A total of 396 individuals 60 years or older with major depression. We conducted model-based analysis to estimate potentially differential intervention effects by education, independent of those by race/ethnicity (and vice versa). INTERVENTION: Algorithm-based recommendations to physicians and care management by care managers. MAIN OUTCOME MEASURES: Antidepressant use, depressive symptoms, and intensity of DCM over 2 years. RESULTS: The PROSPECT intervention had a larger and more lasting effect in less-educated patients. At month 12, the intervention increased the rate of adequate antidepressant use by 14.2 percentage points (pps) (95% confidence interval [CI], 1.7 to 26.4 pps) in the no-college group compared with a null effect in the college-educated group ( 9.2 pps [95% CI, -25.0 to 2.7 pps]); at month 24, the intervention reduced depressive symptoms by 2.6 pps on the Hamilton Depression Rating Scale (95% CI, 4.6 to -0.4 pps) in no-college patients, 3.8 pps (95% CI, -6.8 to -0.4) more than in the college group. The intervention benefitted non-Hispanic white patients more than minority patients. Intensity of DCM received by minorities was 60% to 70% of that received by white patients after the initial phase but did not differ by education. CONCLUSIONS: The PROSPECT intervention substantially reduced disparities by patient education but did not mitigate racial/ethnic disparities in depression treatment and outcomes. Incorporation of culturally tailored strategies in DCM models may be needed to extend their benefits to minorities. TRIAL REGISTRATION: clinicaltrials.gov Identifier for PROSPECT: NCT00279682. PMID- 21646581 TI - Partnering leadership. PMID- 21646582 TI - Poliovirus vaccines: past, present, and future. PMID- 21646583 TI - Effects of a peer-led asthma self-management program for adolescents. AB - OBJECTIVE: To evaluate the effectiveness of a peer-led asthma self-management program for adolescents. DESIGN: Randomized controlled trial comparing a peer-led asthma program (intervention group) and a conventional adult-led asthma program (control group). Each program was implemented at a full-day camp. SETTING: A city and adjacent suburbs in upstate New York. PARTICIPANTS: A total of 112 adolescents aged 13 to 17 years with persistent asthma. INTERVENTION: A peer-led asthma self-management program implemented at a day camp. MAIN OUTCOME MEASURES: The Child Attitude Toward Illness Scale and the Paediatric Asthma Quality of Life Questionnaire were administered at baseline and immediately and 3, 6, and 9 months after the intervention. Spirometry was conducted twice: before and 9 months after the intervention. RESULTS: The intervention group reported more positive attitudes at 6 months (mean difference, 4.11; 95% confidence interval [CI], 0.65-7.56) and higher quality of life at 6 months (difference, 11.38; 95% CI, 0.96-21.79) and 9 months (difference, 12.97; 95% CI, 3.46-22.48) than the control group. The intervention was found to be more beneficial to adolescents of male gender or low family income, as shown by greater improvement in positive attitudes toward asthma and quality of life than their counterparts. CONCLUSION: An asthma self-management program led by peer leaders is a developmentally appropriate approach that can be effective in assisting adolescents with asthma in improving their attitudes and quality of life, particularly for males and those of low socioeconomic status. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT01161225. PMID- 21646585 TI - Community-level determinants of tobacco use disparities in lesbian, gay, and bisexual youth: results from a population-based study. AB - OBJECTIVE: To determine whether characteristics of the social environment surrounding lesbian, gay, and bisexual (LGB) youth contribute to their rates of tobacco use after controlling for established community-level risk factors. DESIGN: Cross-sectional. SETTING: Population-based study of youth. PARTICIPANTS: A total of 31,852 eleventh-grade students (1413 LGB individuals [4.44%]) in Oregon completed the Oregon Healthy Teens survey in 2006-2008. MAIN EXPOSURES: We created a composite index of the social environment in 34 Oregon counties. This measure included the proportion of same-sex couples, the presence of gay-straight alliances in schools, and school policies (nondiscrimination and antibullying) that specifically protected LGB students. MAIN OUTCOME MEASURES: Any tobacco use in the past 30 days. RESULTS: A more supportive social environment for LGB youth was significantly associated with reduced tobacco use (odds ratio, 0.92; 95% confidence interval, 0.90-0.94). This effect remained robust after controlling for sociodemographic variables and multiple community-level risk factors for tobacco use, including median county-level income, exposure to cigarette advertisements, exposure to teacher and peer smoking in schools, and school smoking rules. CONCLUSION: This study documents an association between an objective measure of the social environment and sexual orientation-related disparities in tobacco use. These results highlight the need for structural-level interventions that reduce smoking behaviors in LGB youth. PMID- 21646584 TI - A randomized controlled trial of parental asthma coaching to improve outcomes among urban minority children. AB - OBJECTIVES: To investigate whether asthma coaching decreases emergency department (ED) visits and hospitalizations and increases outpatient asthma monitoring visits. DESIGN: Randomized controlled trial. SETTING: Urban tertiary care children's hospital. PARTICIPANTS: Primary caregivers (hereafter referred to as parents) of children aged 2 to 10 years with asthma who have Medicaid insurance coverage and are urban residents who were attending the ED for acute asthma care. INTERVENTION: Eighteen months of participating in usual care (control group) vs receiving coaching focused on asthma home management, completion of periodic outpatient asthma monitoring visits, and development of a collaborative relationship with a primary care provider (intervention group). MAIN OUTCOME MEASURES: The primary outcome was ED visits. Secondary outcomes were hospitalizations and asthma monitoring visits (nonacute visits focused on asthma care). Outcomes were measured during the year before and 2 years after enrollment. RESULTS: We included 120 intervention parents and 121 control parents. More children of coached parents had at least 1 asthma monitoring visit after enrollment (relative risk [RR], 1.21; 95% confidence interval [CI], 1.04 1.41), but proportions with at least 4 asthma monitoring visits during 2 years were low (20.0% in the intervention group vs 9.9% in the control group). Similar proportions of children in both study groups had at least 1 ED visit (59.2% in the intervention group vs 62.8% in the control group; RR, 0.94; 95% CI, 0.77 1.15) and at least 1 hospitalization (24.2% in the intervention group vs 26.4% in the control group; 0.91; 0.59-1.41) after enrollment. An ED visit after enrollment was more likely if an ED visit had occurred before enrollment (RR, 1.46; 95% CI, 1.16-1.86; adjusted for study group), but risk was similar in both study groups when adjusted for previous ED visits (1.02; 0.82-1.27). CONCLUSION: This parental asthma coaching intervention increased outpatient asthma monitoring visits (although infrequent) but did not decrease ED visits. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00149500. PMID- 21646586 TI - Adolescents' understanding of research concepts: a focus group study. AB - OBJECTIVE: To identify ways to improve adolescents' understanding of informed assent by exploring adolescent comprehension of concepts common to all clinical trials as well as those specific to a human immunodeficiency virus vaccine trial. DESIGN: Qualitative descriptive study. SETTING: Community-based organizations. PARTICIPANTS: Healthy adolescents aged 15 to 17 years in 8 focus groups. INTERVENTION: Focus groups were conducted using a semistructured interview guide. Digital recordings of the groups were transcribed verbatim. OUTCOME MEASURE: Textual data were categorized by 2 investigators using directed qualitative content analysis techniques. Major themes and subthemes were identified, and representative quotes were selected. RESULTS: The general research concepts that were most difficult for teens to understand were placebo and randomization. The most difficult vaccine trial concepts were how a vaccine works and that a vaccine is used for prevention rather than treatment. The most difficult human immunodeficiency virus vaccine-specific trial concept was that standard human immunodeficiency virus antibody tests might provide a false-positive result for participants receiving the test vaccine. Focus group participants wanted to be informed about adverse effects, trial procedures, and whether previous research had been performed before making a decision about trial participation. CONCLUSIONS: Many clinical trial concepts were difficult for teens to understand. Attention needs to be directed toward developing effective ways to explain these concepts to adolescents participating in future human immunodeficiency virus vaccine and other clinical trials. PMID- 21646587 TI - Use of a DASH food group score to predict excess weight gain in adolescent girls in the National Growth and Health Study. AB - OBJECTIVE: To study the effects of selected dietary patterns, particularly a DASH (Dietary Approach to Stop Hypertension) eating pattern, on body mass index (BMI) throughout adolescence. DESIGN: Prospective National Growth and Health Study. SETTING: Washington, DC; Cincinnati, Ohio; and Berkeley, California. PARTICIPANTS: A total of 2327 girls with 10 annual visits starting at age 9 years. MAIN EXPOSURES: Individual DASH-related food groups and a modified DASH adherence score. MAIN OUTCOME MEASURE: The BMI value from measured yearly height and weight over 10 years. RESULTS: Longitudinal mixed modeling methods were used to assess the effects of individual DASH food groups and a DASH adherence score on BMI during 10 years of follow-up, adjusting for race, height, socioeconomic status, television viewing and video game playing hours, physical activity level, and total energy intake. Girls in the highest vs lowest quintile of the DASH score had an adjusted mean BMI of 24.4 vs 26.3 (calculated as weight in kilograms divided by height in meters squared) (P < .05). The strongest individual food group predictors of BMI were total fruit (mean BMI, 26.0 vs 23.6 for <1 vs >=2 servings per day; P < .001) and low-fat dairy (mean BMI, 25.7 vs 23.2 for <1 vs >=2 servings per day; P < .001). Whole grain consumption was more weakly but beneficially associated with BMI. CONCLUSIONS: Adolescent girls whose diet more closely resembled the DASH eating pattern had smaller gains in BMI over 10 years. Such an eating pattern may help prevent excess weight gain during adolescence. PMID- 21646588 TI - Effectiveness of a citywide patient immunization navigator program on improving adolescent immunizations and preventive care visit rates. AB - OBJECTIVE: To assess the impact of a tiered patient immunization navigator intervention (immunization tracking, reminder/recall, and outreach) on improving immunization and preventive care visit rates in urban adolescents. DESIGN: Randomized clinical trial allocating adolescents (aged 11-15 years) to intervention vs standard of care control. SETTING: Eight primary care practices. PARTICIPANTS: Population-based sample of adolescents (N = 7546). INTERVENTION: Immunization navigators at each practice implemented a tiered protocol: immunization tracking, telephone or mail reminder/recall, and home visits if participants remained unimmunized or behind on preventive care visits. MAIN OUTCOME MEASURES: Immunization rates at study end. Secondary outcomes were preventive care visit rates during the previous 12 months and costs. RESULTS: The intervention and control groups were similar at baseline for demographics (mean age, 13.5 years; 63% black, 14% white, and 23% Hispanic adolescents; and 74% receiving Medicaid), immunization rates, and preventive care visit rates. Immunization rates at the end of the study were 44.7% for the intervention group and 32.4% for the control group (adjusted risk ratio, 1.4; 95% confidence interval, 1.3-1.5); preventive care visit rates were 68.0% for the intervention group and 55.2% for the control group (1.2; 1.2-1.3). Findings were similar across practices, sexes, ages, and insurance providers. The number needed to treat for immunizations and preventive care visits was 9. The intervention cost was $3.81 per adolescent per month; the cost per additional adolescent fully vaccinated was $465, and the cost per additional adolescent receiving a preventive care visit was $417. CONCLUSION: A tiered tracking, reminder/recall, and outreach intervention improved immunization and preventive care visit rates in urban adolescents. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00581347. PMID- 21646590 TI - Picture of the month. Subcutaneous fat necrosis of the newborn. PMID- 21646589 TI - Hospital-based comprehensive care programs for children with special health care needs: a systematic review. AB - OBJECTIVE: To examine the effectiveness of hospital-based comprehensive care programs in improving the quality of care for children with special health care needs. DATA SOURCES: A systematic review was conducted using Ovid MEDLINE, CINAHL, EMBASE, PsycINFO, Sociological Abstracts SocioFile, and Web of Science. STUDY SELECTION: Evaluations of comprehensive care programs for categorical (those with single disease) and noncategorical groups of children with special health care needs were included. Selected articles were reviewed independently by 2 raters. DATA EXTRACTION: Models of care focused on comprehensive care based at least partially in a hospital setting. The main outcome measures were the proportions of studies demonstrating improvement in the Institute of Medicine's quality-of-care domains (effectiveness of care, efficiency of care, patient or family centeredness, patient safety, timeliness of care, and equity of care). DATA SYNTHESIS: Thirty-three unique programs were included, 13 (39%) of which were randomized controlled trials. Improved outcomes most commonly reported were efficiency of care (64% [49 of 76 outcomes]), effectiveness of care (60% [57 of 95 outcomes]), and patient or family centeredness (53% [10 of 19 outcomes). Outcomes less commonly evaluated were patient safety (9% [3 of 33 programs]), timeliness of care (6% [2 of 33 programs]), and equity of care (0%). Randomized controlled trials occurred more frequently in studies evaluating categorical vs noncategorical disease populations (11 of 17 [65%] vs 2 of 16 [17%], P = .008). CONCLUSIONS: Although positive, the evidence supporting comprehensive hospital based programs for children with special health care needs is restricted primarily to nonexperimental studies of children with categorical diseases and is limited by inadequate outcome measures. Additional high-quality evidence with appropriate comparative groups and broad outcomes is necessary to justify continued development and growth of programs for broad groups of children with special health care needs. PMID- 21646591 TI - The DASH diet predicts weight gain: what does this mean for my pediatric practice? PMID- 21646592 TI - Increasing adolescent vaccination coverage: the challenges that remain. PMID- 21646593 TI - Hospital-based programs for children with special health care needs: implications for health care reform. PMID- 21646594 TI - Spirometry in biracial children: how adequate are race-based reference equations? PMID- 21646595 TI - Sometimes zero risk is not an option. PMID- 21646596 TI - Advice for patients. Diets to prevent or treat obesity. PMID- 21646597 TI - IkappaBbeta attenuates angiotensin II-induced cardiovascular inflammation and fibrosis in mice. AB - The development of cardiovascular fibrosis is associated with chronic inflammation, where activation of nuclear factor kappaB (NF-kappaB) signaling may play a critical role. NF-kappaB activation is tightly regulated by the cellular inhibitor of kappaB (IkappaB) family of proteins, such as IkappaBalpha and IkappaBbeta. IkappaBalpha and IkappaBbeta display different regulation kinetics in response to inflammatory stimulation. The present study tested the hypothesis that IkappaBalpha and IkappaBbeta may have different roles in modulating cardiovascular inflammation and fibrosis, using a model of angiotensin II infusion-induced hypertension in wild-type mice and IkappaBbeta knock-in mice, in which the IkappaBalpha gene is replaced by IkappaBbeta cDNA (AKBI). In WT mice, subcutaneous angiotensin II infusion for 7 days induced increased perivascular and interstitial collagen deposition and fibrotic lesions, associated with myocardial interstitial hemosiderin accumulation and extensive macrophage infiltration. These effects of angiotensin II were dramatically limited in AKBI mice. Replacement of IkappaBalpha with IkappaBbeta significantly attenuated angiotensin II infusion-induced expression of interleukin 1beta, interleukin 6, monocyte chemotactic protein 1, collagen I and III, fibronectin, and tissue inhibitor of metalloproteinase 1 in the hearts. Furthermore, using cultured vascular smooth muscle cells, we demonstrated that interleukin 1beta-induced NF kappaB activation and monocyte chemotactic protein 1, vascular cell adhesion molecule 1, and tissue inhibitor of metalloproteinase 1 expressions were suppressed in the AKBI cells because of the replacement of IkappaBalpha with IkappaBbeta. These results indicate that NF-kappaB has an essential role in mediating the cardiovascular inflammatory response to angiotensin II and suggest that targeting the balance of IkappaBalpha and IkappaBbeta expression might be a novel therapeutic modality in preventing fibrosis in hypertensive cardiovascular disease. PMID- 21646598 TI - Nongenomic response to aldosterone. PMID- 21646599 TI - beta-Blockers in patients with intermittent claudication and arterial hypertension: results from the nebivolol or metoprolol in arterial occlusive disease trial. AB - The use of beta-receptor blockers in peripheral arterial disease is controversial for their impact on vasomotor tone. The beta-blocker nebivolol possesses vasodilating, endothelium-dependent, NO-releasing properties that might be beneficial in peripheral arterial disease. The aim of the study was to evaluate the effects and tolerability of nebivolol in comparison with metoprolol in these patients. A total of 128 patients with intermittent claudication and essential hypertension were included and double-blind randomized to receive 5 mg of nebivolol (N=65) or 95 mg of metoprolol (N=63) once daily. End points were changes in ankle-brachial index, initial and absolute claudication distance, endothelial function assessed by flow-mediated dilatation of the brachial artery, blood pressure, and quality of life using the claudication scale questionnaire. End point analysis was possible in 109 patients (85.2%). After the 48-week treatment period, ankle-brachial index and absolute claudication distance improved significantly in both patient groups (P<0.05 for both), with no difference across treatments. A significant increase of initial claudication distance was found in the nebivolol group. Adjusted mean change of initial claudication distance was 33.9% after nebivolol (P=0.003) and 16.6% after metoprolol (P=0.12) treatment. Quality of life was not influenced by either treatment, and there was no relevant change in flow-mediated dilatation in patients treated with nebivolol or metoprolol (P=0.16). Both drugs were equally effective in lowering blood pressure. In conclusion, beta-blocker therapy was well tolerated in patients with intermittent claudication and arterial hypertension during a treatment period of ~1 year. In the direct comparison, there was no significant difference between nebivolol and metoprolol. PMID- 21646600 TI - Can we use beta-blocking agents in patients with peripheral artery disease? PMID- 21646601 TI - Sex differences in renal medullary endothelin receptor function in angiotensin II hypertensive rats. AB - We hypothesized that angiotensin (Ang) II hypertensive rats have impaired natriuresis after renal medullary endothelin (ET) B receptor stimulation that would be more evident in male versus female rats. Acute intramedullary infusion of the ET(B) agonist sarafotoxin 6c in normotensive male rats increased sodium excretion from 0.51+/-0.11 MUmol/min during baseline to 1.64+/-0.19 MUmol/min (P<0.05) after S6c. After 2 weeks of Ang II infusion (260 ng/kg per minute SC), male rats had an attenuated natriuretic response to S6c of 0.62+/-0.16 MUmol/min during baseline versus 0.95+/-0.07 MUmol/min after S6c. In contrast, ET(B) dependent natriuresis was similar in female hypertensive rats (0.48+/-0.07 versus 1.5+/-0.18 MUmol/min; P<0.05) compared with normotensive controls (1.05+/-0.07 versus 2.14+/-0.24 MUmol/min; P<0.05). Because ET(A) receptors also mediate natriuresis in normotensive female rats, we examined ET(A) receptor function in female Ang II hypertensive rats. Intramedullary infusion of ET-1 increased sodium excretion in both hypertensive and normotensive female rats, which was partially blocked by the ET(A) antagonist BQ-123. Maximum ET(B) receptor binding in inner medullary membrane preparations was comparable between vehicle and Ang II hypertensive females; however, maximum ET(B) binding was significantly lower in male hypertensive rats (1952+/-251 versus 985+/-176 fmol/mg; P<0.05). These results indicate that renal ET(B) function is impaired in male Ang II hypertension attributed, at least in part, to a reduced number of ET(B) binding sites. Furthermore, renal ET receptor function is preserved in female rats during chronic Ang II infusion, suggesting that renal ET receptor function could serve to limit hypertension in females compared with males. PMID- 21646602 TI - First-line treatment for primary testicular diffuse large B-cell lymphoma with rituximab-CHOP, CNS prophylaxis, and contralateral testis irradiation: final results of an international phase II trial. AB - PURPOSE: Primary testicular lymphoma (PTL) has poor prognosis with failures in contralateral testis, CNS, and extranodal sites. To prevent these events, we designed an international phase II trial (International Extranodal Lymphoma Study Group 10 [IELSG-10]) that addressed feasibility and activity of conventional chemoimmunotherapy associated with CNS prophylaxis and contralateral testis irradiation. The trial was conducted by the IELSG and the Italian Lymphoma Foundation. PATIENTS AND METHODS: Fifty-three patients (age 22 to 79 years) with untreated stage I or II PTL were treated with six to eight courses of rituximab added to cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) every 21 days (R-CHOP21); four doses of intrathecal methotrexate (IT-MTX) and radiotherapy (RT) to the contralateral testis (30 Gy) for all patients and to regional lymph nodes (30 to 36 Gy) for stage II disease. RESULTS: All patients received R-CHOP21, 50 received CNS prophylaxis, and 47 received testicular RT. With a median follow-up of 65 months, 5-year progression-free survival and overall survival rates were 74% (95% CI, 59% to 84%) and 85% (95% CI, 71% to 92%), respectively. Ten patients relapsed or progressed: two in lymph nodes, five in extranodal organs, and three in the CNS. The 5-year cumulative incidence of CNS relapse was 6% (95% CI, 0% to 12%). No contralateral testis relapses occurred. Ten patients died: lymphoma (n = 6), secondary leukemia (n = 2), heart failure (n = 1), and gastric cancer (n = 1). Grade 3 to 4 toxicities were neutropenia, 28%; infections, 4%; and neurologic, 13%. No deaths occurred as a result of toxicity. CONCLUSION: This international prospective trial shows that combined treatment with R-CHOP21, IT-MTX, and testicular RT was associated with a good outcome in patients with PTL. RT avoided contralateral testis relapses, but CNS prophylaxis deserves further investigation. PMID- 21646603 TI - Symptom scores and delirium in advanced cancer. PMID- 21646605 TI - Impact of KRAS and BRAF gene mutations on targeted therapies in colorectal cancer. PMID- 21646604 TI - Impact of young age on treatment efficacy and safety in advanced colorectal cancer: a pooled analysis of patients from nine first-line phase III chemotherapy trials. AB - PURPOSE: Colorectal cancer predominantly occurs in the elderly, but approximately 5% of patients are 50 years old or younger. We sought to determine whether young age is prognostic, or whether it influences efficacy/toxicity of chemotherapy, in patients with advanced disease. METHODS: We analyzed individual data on 6,284 patients from nine phase III trials of advanced colorectal cancer (aCRC) that used fluorouracil-based single-agent and combination chemotherapy. End points included progression-free survival (PFS), overall survival (OS), response rate (RR), and grade 3 or worse adverse events. Stratified Cox and adjusted logistic regression models were used to test for age effects and age-treatment interactions. RESULTS: A total of 793 patients (13%) were younger than 50 years old; 188 of these patients (3% of total patients) were younger than 40 years old. Grade 3 or worse nausea (10% v 7%; P = .01) was more common, and severe diarrhea (11% v 14%; P = .001) and neutropenia (23% v 26%; P < .001) were less common in young (younger than 50 years) than in older (older than 50 years) patients. Age was prognostic for PFS, with poorer outcomes occurring in those younger than 50 years (median, 6.0 v 7.5 months; hazard ratio, 1.10; P = .02), but it did not affect RR or OS. In the subset of monotherapy versus combination chemotherapy trials, the relative benefits of multiagent chemotherapy were similar for young and older patients. Results were comparable when utilizing an age cut point of 40 years. CONCLUSION: Young age is modestly associated with poorer PFS but not OS or RR in treated patients with aCRC, and young patients have more nausea but less diarrhea and neutropenia with chemotherapy in general. Young versus older patients derive the same benefits from combination chemotherapy. Absent results of a clinical trial, standard combination chemotherapy approaches are appropriate for young patients with aCRC. PMID- 21646606 TI - Nicotinic acetylcholine receptor polymorphism, smoking behavior, and tobacco related cancer and lung and cardiovascular diseases: a cohort study. AB - PURPOSE: We examined the associations between the nicotinic acetylcholine receptor polymorphism (rs1051730) on chromosome 15q25 marking the gene cluster CHRNA3-CHRNB4-CHRNA5, smoking behavior, and tobacco-related cancer and lung and cardiovascular diseases in the general population. METHODS: Ten thousand three hundred thirty participants from the Copenhagen City Heart Study were genotyped and observed prospectively with up to 18 years of 100% complete follow-up. Smoking behavior was measured at baseline. End points were lung cancer, bladder cancer, chronic obstructive pulmonary disease, ischemic heart disease, and ischemic stroke. RESULTS: Multifactorially adjusted and genotype-adjusted subhazard ratios for a cumulative tobacco consumption above 40 pack-years versus 0 pack-years were 32.5 (95% CI, 12.0 to 87.7) for lung cancer, 2.2 (95% CI, 1.1 to 4.5) for bladder cancer, 9.4 (95% CI, 6.9 to 12.7) for chronic obstructive pulmonary disease, 1.5 (95% CI, 1.3 to 1.8) for ischemic heart disease, and 1.1 (95% CI, 0.8 to 1.4) for ischemic stroke. Among smoking noncarriers and homozygotes, daily tobacco consumption was 16 and 18 g/d (P < .001), cumulative tobacco consumption was 28 and 31 pack-years (P = .003), and smoking inhalation was 71.9% and 78.1% (P < .001), respectively. Multifactorially adjusted and smoking behavior-adjusted subhazard ratios for homozygotes versus noncarriers were 1.6 (95% CI, 1.1 to 2.2) for lung cancer, 1.7 (95% CI, 1.0 to 3.0) for bladder cancer, 1.3 (95% CI, 1.1 to 1.6) for chronic obstructive pulmonary disease, 0.9 (95% CI, 0.7 to 1.0) for ischemic heart disease, and 1.1 (95% CI, 0.8 to 1.4) for ischemic stroke. CONCLUSION: Although smoking is associated with major tobacco-related diseases in the general population, the nicotinic acetylcholine receptor polymorphism is associated with additional increased risk of lung cancer, bladder cancer, and chronic obstructive pulmonary disease after adjustment for smoking. PMID- 21646607 TI - Splenic gas as a result of a non-Hodgkin's lymphoma in a patient with common variable immunodeficiency. PMID- 21646608 TI - New initiatives at Journal of Clinical Oncology. PMID- 21646609 TI - Isolated CNS relapse in human epidermal growth factor receptor 2-positive esophagogastric cancer: effective treatment with trastuzumab after failure of surgery and radiotherapy. PMID- 21646610 TI - Newly diagnosed acute promyelocytic leukemia: arsenic moves front and center. PMID- 21646611 TI - Radiation recall 5 years after whole-breast irradiation for early-stage breast cancer secondary to initiation of rosuvastatin and amlodipine. PMID- 21646612 TI - Ras inhibition and the survival benefit favoring zoledronic acid compared with denosumab in patients with multiple myeloma. PMID- 21646613 TI - Successful treatment of a patient with Epstein-Barr virus-positive B-cell lymphoproliferative disorder resembling post-transplant lymphoproliferative disorder using single-agent rituximab. PMID- 21646614 TI - Trustworthiness. PMID- 21646615 TI - Phase II study of single-agent arsenic trioxide for the front-line therapy of acute promyelocytic leukemia. AB - PURPOSE: The long-term follow-up results of patients with acute promyelocytic leukemia (APL) treated with all-trans retinoic acid and chemotherapy show high cure rates. Several studies have shown high efficacy of single-agent arsenic trioxide in newly diagnosed APL. However, long-term follow-up results are needed. PATIENTS AND METHODS: One hundred ninety-seven patients with newly diagnosed APL were treated with arsenic trioxide 0.15 mg/kg daily intravenous infusion until complete remission (CR). After achieving CR, the patients received one to four more courses of therapy with arsenic trioxide as consolidation and were observed with reverse-transcriptase polymerase chain reaction studies from peripheral blood (to detect of minimal residual disease) every 3 months or until relapse or death. RESULTS: The morphologic CR rate was 85.8%. The most common cause of remission failure was early death owing to APL differentiation syndrome (13.2%). The most important prognostic factor for early mortality was a high WBC count at presentation. The 5-year disease-free survival (DFS) rate was 66.7% +/- 4% (SE). Relapse after 5 years in CR was rare. The 5-year overall survival (OS) rate by intention-to-treat analysis was 64.4% +/- 4%. In patients who achieved CR, OS and DFS were identical. CONCLUSION: The long-term follow-up of newly diagnosed patients with APL treated with single-agent arsenic trioxide shows high rates of DFS and OS. PMID- 21646616 TI - Impact of KRAS and BRAF Gene Mutation Status on Outcomes From the Phase III AGITG MAX Trial of Capecitabine Alone or in Combination With Bevacizumab and Mitomycin in Advanced Colorectal Cancer. AB - PURPOSE: Mutations affecting the KRAS gene are established predictive markers of outcome with anti-epithelial growth factor receptor (EGFR) antibodies in advanced colorectal cancer (CRC). The relevance of these markers for anti-vascular endothelial growth factor (VEGF) therapy is controversial. This analysis was performed to assess the predictive and prognostic impact of KRAS and BRAF gene mutation status in patients receiving capecitabine with bevacizumab (CG) or capecitabine without bevacizumab in the phase III AGITG MAX (Australasian Gastrointestinal Trials Group MAX) study. PATIENTS AND METHODS: Mutation status was determined for 315 (66.9%) of the original 471 patients. Mutation status was correlated with efficacy outcomes (response rate, progression-free survival [PFS], and overall survival [OS]), and a predictive analyses was undertaken. RESULTS: Mutations in KRAS and BRAF genes were observed in 28.8% and 10.6% of patients, respectively. KRAS gene mutation status (wild type [WT] v mutated [MT]) had no prognostic impact for PFS (hazard ratio [HR], 0.89; CI, 0.69 to 1.14) or OS (HR, 0.97; CI, 0.73 to 1.28). BRAF mutation status (WT v MT) was not prognostic for PFS (HR, 0.80; CI, 0.54 to 1.18) but was prognostic for OS (HR, 0.49; CI, 0.33 to 0.73; P = .001). By using the comparison of capecitabine versus capecitabine and bevacizumab (CB) and CB plus mitomycin (CBM), KRAS gene mutation status was not predictive of the effectiveness of bevacizumab for PFS or OS (test for interaction P = .95 and 0.43, respectively). Similarly, BRAF gene mutation status was not predictive of the effectiveness of bevacizumab for PFS or OS (test for interaction P = .46 and 0.32, respectively). CONCLUSION: KRAS gene mutation status was neither prognostic for OS nor predictive of bevacizumab outcome in patients with advanced CRC. BRAF gene mutation status was prognostic for OS but was not predictive of outcome with bevacizumab. PMID- 21646617 TI - Granular cell tumor of the breast: clinical and pathologic characteristics of a rare case in a 14-year-old girl. PMID- 21646618 TI - Case of rhabdomyosarcoma presenting with myasthenia gravis. PMID- 21646619 TI - Teaching Video NeuroImages: epilepsy with myoclonic absences: a distinct electroclinical syndrome. PMID- 21646620 TI - On the 60th anniversary of Neurology(r). PMID- 21646621 TI - Fluctuating concepts of childhood absence epilepsy. PMID- 21646622 TI - Resting functional connectivity between the hemispheres in childhood absence epilepsy. AB - OBJECTIVE: The fundamental mechanisms by which childhood absence epilepsy (CAE) changes neural networks even between seizures remain poorly understood. During seizures, cortical and subcortical networks exhibit bihemspheric synchronous activity based on prior EEG-fMRI studies. Our aim was to investigate whether this abnormal bisynchrony may extend to the interictal period, using a blood oxygen level-dependent (BOLD) resting functional connectivity approach. METHODS: EEG fMRI data were recorded from 16 patients with CAE and 16 age- and gender-matched controls. Three analyses were performed. 1) Using 16 pairs of seizure-related regions of interest (ROI), we compared the between-hemisphere interictal resting functional connectivity of patients and controls. 2) For regions showing significantly increased interhemispheric connectivity in CAE, we then calculated connectivity to the entire brain. 3) A paired-voxel approach was performed to calculate resting functional connectivity between hemispheres without the constraint of predefined ROIs. RESULTS: We found significantly increased resting functional connectivity between hemispheres in the lateral orbitofrontal cortex of patients with CAE compared to normal controls. Enhanced between-hemisphere connectivity localized to the lateral orbitofrontal cortex was confirmed by all 3 analysis methods. CONCLUSIONS: Our results demonstrate abnormal increased connectivity between the hemispheres in patients with CAE in seizure-related regions, even when seizures were not occurring. These findings suggest that the lateral orbitofrontal cortex may play an important role in CAE pathophysiology, warranting further investigation. In addition, resting functional connectivity analysis may provide a promising biomarker to improve our understanding of altered brain function in CAE during the interictal period. PMID- 21646623 TI - Etiologic investigation of ischemic stroke in young adults. AB - OBJECTIVES: We attempted to classify causes of ischemic stroke in young adults using a progressive diagnostic algorithm and the ASCO (atherosclerosis, small vessel disease, cardiac source, other cause) classification system. METHODS: Patients aged 16-54 years consecutively treated for acute ischemic stroke in a tertiary stroke unit were included in this retrospective analysis. Causes of stroke were classified using the ASCO system, which assigns a graded level of likelihood to each potential cause in individual patients. The initial etiologic workup included brain imaging, magnetic resonance or CT angiography of cerebral and cervical vessels, EKG, and routine blood studies. Patients without a definite cause of ischemic stroke after initial evaluation underwent transesophageal echocardiography. RESULTS: We included 318 patients (195 men and 123 women); 131 patients were aged 16-44 years, and 187 were aged 45-54 years. A definite cause of stroke (ASCO grade 1) could be identified in 145 patients (45.5%). An uncertain cause of stroke (ASCO grade 2) was found in 59 (18.5%) further patients. Most (130 of 145) definite causes were identified by initial evaluation. The 2 major definite or uncertain causes of stroke were patent foramen ovale associated with atrial septal aneurysm (PFO-ASA) (20 of 131 [15.3%]) and dissection of the cervical or cerebral artery (19 of 131 [14.5%]) in patients aged 16-44 years and large-vessel atherosclerosis (37 of 187 [19.8%]) and PFO-ASA (23 of 187 [12.3%]) in patients aged 45-54 years. CONCLUSIONS: Our findings suggest that PFO-ASA may be a major cause of ischemic stroke in young adults. PMID- 21646624 TI - Common viruses associated with lower pediatric multiple sclerosis risk. AB - BACKGROUND: Because common viruses are encountered during childhood, pediatric multiple sclerosis (MS) offers a unique opportunity to investigate the influence of these viruses on disease susceptibility and the interactions between seroprevalence and select HLA genotypes. We studied seroprevalence for Epstein Barr virus (EBV), cytomegalovirus (CMV), and herpes simplex virus (HSV) type 1 and HLA-DRB1*1501/1503 status as predictors of pediatric MS. METHODS: This was a retrospective analysis of prospectively collected demographic, clinical, and biologic data in subjects up to 18 years of age with early MS, control subjects seen at the same regional referral pediatric MS clinics, and additional healthy pediatric control subjects. RESULTS: Patients with early pediatric MS (n=189) and pediatric control subjects (n=66) were tested. Epstein-Barr nuclear antigen-1 seropositivity was associated with an increased odds of MS (odds ratio [OR] 3.78, 95% confidence interval [CI] 1.52-9.38, p=0.004) in analyses adjusted for age, sex, race, ethnicity, and HLA-DRB1*1501/1503 status. In multivariate analyses including EBV status, a remote infection with CMV (OR 0.27, 95% CI 0.11-0.67, p=0.004) was associated with a lower risk of developing MS. Although a remote infection with HSV-1 was not associated with an increased odds of MS, a strong interaction was found between HSV-1 status and HLA-DRB1 in predicting MS (p<0.001). HSV-1 was associated with an increased risk of MS in those without a DRB1*15 allele (OR 4.11, 95% CI 1.17-14.37, p=0.03), whereas the effect was reversed in those who were DRB1*15-positive (OR 0.07, 95% CI 0.02-0.32, p=0.001). CONCLUSIONS: These findings suggest that some infections with common viruses may in fact lower MS susceptibility. If this is confirmed, the pathways for risk modification remain to be elucidated. PMID- 21646626 TI - Demographic and clinical characteristics of malignant multiple sclerosis. AB - OBJECTIVE: Multiple sclerosis (MS) that causes patients to require assistance for ambulation (Expanded Disability Status Scale [EDSS] >=6) within 5 years from symptom onset is generally termed malignant. Malignant status can be transient (TM) or sustained until year 5 (SM). We studied the incidence, predictors, and demographic and clinical characteristics of malignant MS. METHODS: Patients with symptom onset in 2002-2005 and 5-year follow-up were selected from the Partners Multiple Sclerosis Center database. Patients with TM were further grouped into TM and SM. The mechanism of reaching EDSS 6 (relapse- vs progression-related) was determined. RESULTS: A total of 487 patients were included (17 TM, 42 SM). The incidence proportion of ever malignant (EM=SM+TM) was estimated as 12.11% and SM as 8.62%. Patients with older age at onset, male gender, and positive smoking history were more likely to become SM. Compared to nonmalignant patients, the proportion of progressive-onset MS in the SM group was significantly higher, but not different in TM. Within relapsing-onset patients, most of TM, and a smaller proportion of the SM group had a relapse-related as opposed to progression related mechanism. The final model predictors for EM vs nonmalignant were older age at onset, motor symptoms at onset, and progressive disease onset. Within the malignant patients, predictors of TM vs SM were younger age and brainstem symptoms at onset. CONCLUSIONS: Over 10% of patients with MS experience a malignant course as defined above. Some demographic and clinical factors are found to predict a malignant outcome. MS in patients who reach a high EDSS based on disease progression is more likely to remain malignant. PMID- 21646627 TI - Meta-analysis of the relationship between Parkinson disease and melanoma. AB - OBJECTIVE: To assess the epidemiologic evidence on melanoma in relation to Parkinson disease (PD) via systematic review and meta-analysis. METHODS: Epidemiologic studies on melanoma and PD were searched using PubMed, Web of Science, Scoups, and Embase (1965 through June 2010). Eligible studies were those that reported risk estimates of melanoma among patients with PD or vice versa. Pooled odds ratios (ORs) with 95% confidence intervals (CIs) were calculated using random-effects models. RESULTS: We identified 12 eligible publications on melanoma and PD: 8 had fewer than 10 cases with both PD and melanoma, and 7 provided gender-specific results. The pooled OR was 2.11 (95% CI 1.26-3.54) overall, 2.04 (1.55-2.69) for men, and 1.52 (0.85-2.75) for women. Analyses by temporal relationship found that melanoma occurrence was significantly higher after the diagnosis of PD (OR 3.61, 95% CI 1.49-8.77), but not before PD diagnosis (OR 1.07, 95% CI 0.62-1.84). Further analyses revealed that the lack of significance in the latter analysis was due to one study, which when excluded resulted in a significant association (OR 1.44, 95% CI 1.06-1.96). We also analyzed nonmelanoma skin cancers in relation to PD and found no significant relationship (OR 1.11, 95% CI 0.94-1.30). CONCLUSIONS: Collective epidemiologic evidence supports an association of PD with melanoma. Further research is needed to examine the nature and mechanisms of this relationship. PMID- 21646629 TI - Assessment of sympathetic index from the Valsalva maneuver. AB - OBJECTIVE: Blood pressure (BP) decline and recovery during the Valsalva maneuver (VM) are used to evaluate the degree of sympathetic failure (SF) but a reliable sympathetic index (SI) derived from VM is lacking. METHODS: Patients with mild (n=20), moderate (n=65), and severe (n=60) SF and 23 healthy controls were evaluated using a standardized battery of autonomic tests. SF was defined as mild (associated with reduced sudomotor volumes at distal leg); moderate (associated with a fall in systolic BP >=10< 30 mm Hg during the tilt test); and severe (associated with a fall in systolic BP >=30 mm Hg during the tilt test). Six SIs were compared: SI1 (BP fall during phase 2), SI2 (BP recovery during phase 2), SI3 (the difference in BP between baseline and the end of phase 2), SI4 (the magnitude of phase 4), SI5 (BP recovery time), and SI6 (baroreflex sensitivity index). RESULTS: All indexes showed overall significant differences among tested groups (p<0.05). Only SI3 differentiated all subject groups. Compared to other SIs, SI3 correlated the most with orthostatic hypotension (OH; r=0.62, p < 0.05) during the tilt. CONCLUSIONS: SI3 is the optimal method for calculation of SI since it 1) easily differentiates between healthy controls and those with SF; 2) correlates with the OH, a proxy for a sympathetic failure; 3) tracks the full spectrum of SF (mild-moderate-severe). SI3 expands the utility of quantitative autonomic testing. PMID- 21646630 TI - Small-fiber neuropathy in patients with ALS. AB - OBJECTIVE: To investigate the involvement of the epidermal small sensory fibers in the neurodegenerative process in amyotrophic lateral sclerosis (ALS). METHODS: In the present study, skin biopsies of 28 patients with ALS were obtained at an average of 34 months after disease onset by history. Protein gene product 9.5 (PGP9.5) immunohistochemistry findings were compared to 17 age-matched controls. The primary endpoint of the study was to evaluate the decrease in the density of small intraepidermal nerve fibers and to compare the prevalence of small-fiber neuropathy in patients with ALS and in controls. RESULTS: We found a significant reduction in epidermal nerve fiber density in the distal calf of patients with ALS (4.8 +/- 3.7 fibers/mm vs 12.2 +/- 4.6 in age-matched controls, p<0.0001). The extent of fiber loss was age-dependent. Also, the number of subjects with small-fiber neuropathy was significantly higher in the ALS group than in the controls (79% vs 12%). Correspondingly, mild sensory symptoms including diffuse dysesthesias, paresthesias, and hypesthesia were found in 7 patients. In 17 biopsies of patients with ALS, but only in 2 controls, we saw larger (>1.5 MUm in diameter) focal swellings of epidermal axons resembling spheroids, suggesting trafficking defects. CONCLUSIONS: These results indicate that small, distal epidermal nerve fibers are involved in this disease, supporting the concept of distal axonopathy in ALS. PMID- 21646631 TI - ALS risk but not phenotype is affected by ataxin-2 intermediate length polyglutamine expansion. PMID- 21646632 TI - RRM2B mutations are frequent in familial PEO with multiple mtDNA deletions. PMID- 21646633 TI - Whole-body imaging in schwannomatosis. PMID- 21646634 TI - Evidence-based guideline: the role of diffusion and perfusion MRI for the diagnosis of acute ischemic stroke: report of the Therapeutics and Technology Subcommittee of the American Academy of Neurology. PMID- 21646636 TI - Development and psychometric testing of the colonoscopy embarrassment scale. AB - Colorectal cancer, the third leading cause of cancer-related death in the United States, could largely be prevented if more people had polyps removed via colonoscopies. Embarrassment is one important barrier to colonoscopy, but little is known about embarrassment in this context, and there were no reliable and valid measures of this construct. The purpose of this study was to develop a reliable and valid instrument to measure colonoscopy-related embarrassment. Transtheoretical Model of Behavior Change and Health Belief Model provided the theoretical basis for this study. Participants were health maintenance organization members aged 50 to 65 years (N=234). Using a cross-sectional, descriptive research design, data were collected using a mailed survey. Internal consistency (Cronbach's alpha=.96) and construct validity of the 13-item instrument were demonstrated. This unidimensional scale shows promise as a valid and reliable instrument to measure colonoscopy-related embarrassment and to inform development of interventions to reduce embarrassment, leading to higher colonoscopy completion rates and lower mortality. PMID- 21646637 TI - Treating persistent distress and anxiety in parents of children with cancer: an initial feasibility trial. AB - Persistent anxiety is common among parents of children with cancer and may affect the family's well-being and adjustment. The goals of this pilot study are to determine the feasibility and potential efficacy of a brief cognitive-behavioral parent intervention aimed at reducing parental distress and anxiety related to their child's cancer diagnosis. Parents of children with cancer, at least 1 month postdiagnosis, were screened at an outpatient oncology clinic, and those reporting elevated levels of distress were offered a 4-session cognitive behavioral intervention based on a modified version of the Surviving Cancer Competently Intervention Program-Newly Diagnosed. Five parents reporting persistent distress received the intervention. Results revealed decreases in parents' distress, state anxiety, and depressive symptoms, as well as in parents' feelings of burden associated with their children's cancer. This initial study suggests that identification of parents with prolonged heightened psychological distress is feasible and acceptable and that offering them a brief intervention within a pediatric oncology setting may be beneficial. PMID- 21646638 TI - Cultural beliefs and coping strategies related to childhood cancer: the perceptions of South Asian immigrant parents in Canada. AB - The purpose of this study is to describe cultural beliefs and coping strategies related to dealing with childhood cancer identified through a qualitative study of the caregiving experiences of first-generation South Asian immigrant parents of children with cancer. A constructivist grounded theory approach was employed. Families with a child at least 6 months postdiagnosis were recruited from 5 Canadian pediatric oncology centers. In-depth semistructured interviews were conducted in English, Hindi, Punjabi, or Urdu with a sample of 25 South Asian parents. Analysis of interviews involved line-by-line coding and using the constant comparison method. The following 2 central themes related to culture and coping emerged: (a) cultural beliefs about childhood cancer being incurable, rare, unspeakable, and understood through religion and (b) parental coping strategies included gaining information about the child's cancer, practicing religious rituals and prayers, trusting the health care professionals, and obtaining mutual support from other South Asian parents. These cultural beliefs and coping strategies have important implications for health care providers to understand the variations in the perceptions of childhood cancer and coping in order to implement culturally sensitive health care services. PMID- 21646639 TI - The camp experience for siblings of pediatric cancer patients. AB - Siblings of pediatric cancer patients experience difficulties coping and adapting to the experience of a cancer diagnosis. A variety of emotional and behavioral changes as well as somatic complaints have been reported. Children describe many negative changes after their sibling is diagnosed with cancer. Many social supports and therapeutic interventions have been proposed for siblings, one of which is a camp experience. The literature has demonstrated that camps have a positive impact and offer siblings of children with cancer a supportive peer environment. Camp encourages discussion with peers and health care providers and facilitates participation in activities that improve knowledge, social confidence, and self-esteem. Nurses can help siblings by recommending camp experiences, volunteering at camps, and adding a camp experiences to existing sibling support programs. PMID- 21646640 TI - Tp63 in oral development, neoplasia, and autoimmunity. AB - The Tp63 gene encodes for multiple isoforms of the p63 transcription factor, a member of the p53 family of proteins. Much like its more famous sibling, the biological role of p63 is quite complex, with wide-ranging effects on development, differentiation, and cellular lineage choices. The crucial function of p63 is epitomized by the striking phenotype of p63 knockout mice. These animals have a profound block in the development of stratified epithelia and aplasia of multiple ectodermal appendages, as well as orofacial clefting and limb defects. Remarkably, a similar spectrum of phenotypic alterations is observed in human syndromes resulting from Tp63 gene mutations. p63 is an important hub in the transcriptional and signaling networks of epithelial cells; thus, it is not surprising that dysregulation of this transcription factor is associated with squamous cell carcinoma. Finally, as a testament to the growing repertoire of p63 associated diseases, autoantibodies to p63 are associated with chronic ulcerative stomatitis, an oral immunologically mediated disease. Over the past decade, our understanding of the broad biologic and pathophysiological roles of p63 has grown significantly. In this review, we discuss the molecular attributes of Tp63 and the clinical consequences of Tp63 dysregulation, particularly as it pertains to oral tissues. PMID- 21646644 TI - Recurrent cardiovascular events with paclitaxel-eluting versus bare-metal stents in saphenous vein graft lesions: insights from the SOS (Stenting of Saphenous Vein Grafts) trial. AB - The Stenting of Saphenous Vein Grafts (SOS) trial demonstrated a reduction in clinical and angiographic adverse events with paclitaxel-eluting stents (PES) compared to bare-metal stents (BMS) in saphenous vein graft (SVG) lesions, but the rate of recurrent adverse events has not been described. METHODS: We performed a post hoc, landmark analysis to evaluate the risk of event recurrence following a non-fatal initial event among the SOS trial patients (pts). RESULTS: During a median follow-up of 35 months, the 80 pts enrolled in SOS experienced a total of 78 major cardiovascular events (MACE): 51 in the BMS group and 27 in PES group. No MACE were found in 28 pts (35%) while 52 pts (65%) had at least one event. The initial event was death in 13 pts (16%). Among the 39 pts whose initial event was not fatal, 12 (31%) had one or more subsequent MACE (50% of which were definitely related to the study SVG). The mean and median number of MACE per patient was significantly higher in patients receiving BMS versus PES (1.3 +/- 1.2 and 1 +/- 1.26 versus 0.6 +/- 0.7 and 1 +/- 0.825, p = 0.005 and p = 0.008, respectively). The rate of a second MACE following an initial event was 17% in the PES group and 37% in the BMS group (p = 0.24). Ten of 12 pts with recurrent events had received a BMS (83%). CONCLUSION: Pts undergoing SVG stenting had a high rate of recurrent events after an initial non-fatal event. These events were often related to the target vessel and most occurred in pts who had received a BMS, further supporting the benefit of PES over BMS in SVG lesions. PMID- 21646642 TI - Unraveling lactococcal phage baseplate assembly by mass spectrometry. AB - Bacteriophages belonging to the Caudovirales order possess a tail acting as a molecular machine used during infection to recognize the host and ensure high efficiency genome delivery to the cell cytoplasm. They bear a large and sophisticated multiprotein organelle at their distal tail end, either a baseplate or a tail-tip, which is the control center for infectivity. We report here insights into the baseplate assembly pathways of two lactoccocal phages (p2 and TP901-1) using electrospray ionization-mass spectrometry. Based on our "block cloning" strategy we have expressed large complexes of their baseplates as well as several significant structural subcomplexes. Previous biophysical characterization using size-exclusion chromatography coupled with on-line light scattering and refractometry demonstrated that the overproduced recombinant proteins interact with each other to form large (up to 1.9 MDa) and stable assemblies. The structures of several of these complexes have been determined by x-ray diffraction or by electron microscopy. In this contribution, we demonstrate that electrospray ionization-mass spectrometry yields accurate mass measurements for the different baseplate complexes studied from which their stoichiometries can be discerned, and that the subspecies observed in the spectra provide valuable information on the assembly mechanisms of these large organelles. PMID- 21646645 TI - Stenting of saphenous vein grafts--a treacherous road to travel. PMID- 21646646 TI - Prospective, randomized, placebo-controlled evaluation of esomeprazole in coronary artery disease patients. EPAC: esomeprazole prevention of atypical chest pains. AB - BACKGROUND: Coronary artery disease (CAD) patients experience chest pain (CP) from cardiac and non-cardiac etiologies. OBJECTIVE: Proton pump inhibitor (PPI) treatment of CAD patients with atypical CP would result in less CP, as well as fewer emergency room (ER) visits for CP, medical evaluations, and hospitalizations. METHODS: A randomized, prospective, placebo-controlled trial in patients with a history of severe CAD, without current ischemia, and not receiving acid reducing therapy. Patients received esomeprazole 40 mg or placebo daily added to standard cardiac medications for 6 months. The primary endpoint was percentage of patients experiencing CP. RESULTS: In 162 patients randomized, 78 received esomeprazole and 84 received placebo. Esomeprazole significantly reduced the primary endpoint of patients experiencing CP (24.4% versus 54.8%; p < 0.001). The absolute number of CP episodes and CP days was also significantly reduced. Fewer patients experienced the combined endpoint of doctor office/ER visits for CP (19.2% versus 48.8%; p < 0.001), mainly due to significantly fewer office visits for CP (14.1% versus 39.3%; p < 0.001). ER visits for CP trended less with esomeprazole (12.8% versus 22.6%; p = 0.10), as did hospitalizations for CP (14.1% versus 20.2%; p = 0.30). However, the study lacked statistical power to reach these secondary endpoints. CONCLUSIONS: Concomitant esomeprazole therapy in CAD patients without active ischemia but with a history of atypical CP reduces number of patients with further CP symptoms and medical office visits for CP. Larger studies are needed to further evaluate PPI treatment on ER CP visits and hospitalizations in this patient population. PMID- 21646647 TI - Renal artery plaque composition is associated with changes in renal frame count following renal artery stenting. AB - BACKGROUND: Benefit of percutaneous revascularization for atherosclerotic renal artery stenosis (RAS) may be attenuated by distal embolization of atheroemboli. The purpose of this study was to characterize RAS plaque composition with intravascular ultrasound virtual histology (IVUS-VH) and to explore the relationship between plaque components and renal frame count (RFC) after renal revascularization. METHODS: Seventeen patients (75 +/- 7.5 years; 18 lesions) undergoing RAS revascularization were included. Before stenting, automated IVUS VH pullback (0.5 mm/sec) with analysis of the minimal luminal diameter (MLD) frame and entire atherosclerotic segment was performed. RFC was also determined before and after stenting. RESULTS: The VH component analysis of the segment demonstrated predominantly fibrous tissue (56.3 +/- 11.4%), followed by necrotic core (21.8 +/- 8.6%), dense calcification (13.2 +/- 6.6%) and fibrofatty tissue (8.7 +/- 4.0%). Analysis of the MLD frame also demonstrated mostly fibrous tissue (62.1 +/- 11.1%), with smaller amounts of necrotic core (15.6 +/- 7.3%), fibrofatty (13.9 +/- 9.6%), and dense calcification (8.4 +/- 6.0%). A trend toward more fibrous tissue (p = 0.074), less necrotic core (p = 0.095) and less dense calcification (p = 0.075) at the MLD compared to the segment was observed. Analysis of the entire atherosclerotic segment revealed a positive correlation between % necrotic core and change in RFC (r = 0.582; p = 0.029), with increasing necrotic core associated with an increase in RFC after revascularization. CONCLUSION: Both the MLD frame and segmental analysis of atherosclerotic RAS lesions demonstrate predominantly fibrous tissue with smaller amounts of necrotic core, fibrofatty tissue, and dense calcification. Increased necrotic core correlates with a lack of improvement in RFC after stenting. PMID- 21646648 TI - Renal arterial ultrasound--predictor of things to come? PMID- 21646649 TI - Pericardiocentesis in contemporary practice. AB - OBJECTIVE: Pericardiocentesis is a life-saving procedure associated with a small, but significant, risk of major complication. An apical or subcostal approach may be used, although the relative complication rates are not reported. In modern practice, an increasing proportion of pericardial effusions occur as a result of catheter-laboratory related complications. This study examines current practice and analyzes the complications of pericardial drainage according to the route of approach. DESIGN: Historical cohort study. SETTING: Four Oxfordshire hospitals, including the John Radcliffe Hospital, a tertiary referral center. PATIENTS: Local databases were searched to identify percutaneous pericardiocenteses carried out between November 2002 and October 2009. RESULTS: A total of 188 pericardiocenteses were performed in 163 patients. Malignancy (55; 33.7%) and catheter-based cardiac procedures (45; 23.9%) were the most common causes of pericardial effusions requiring drainage. 50.0% of all pericardiocenteses were performed in patients who had received anticoagulant or antiplatelet agents the same day. This rose to 93.7% in patients whose effusions occurred as a complication of a catheter-based procedure. Nine complications occurred during the study period, giving an overall complication rate of 4.8%. Six of the complications occurred via the subcostal route and all 4 complications requiring surgery occurred via the subcostal route. CONCLUSION: The numbers of iatrogenic pericardial effusions occurring as a complication of catheter-based procedures mean that a significant proportion of pericardiocenteses are being performed in anticoagulated patients. This may alter the risk profile. Although complication rates were low for both routes, all major complications requiring surgery occurred via the subcostal approach. These data suggest an apical approach may be preferable where practical. PMID- 21646650 TI - Elevated B-type natriuretic peptide levels in patients undergoing coronary stenting. AB - BACKGROUND: Release of B-type natriuretic peptide (BNP) may be triggered by tissue hypoxia even in the absence of left ventricular (LV) dysfunction, generating interest in studying changes in levels following percutaneous coronary intervention (PCI). Though the prognostic role of natriuretic peptides following elective PCI has been documented, most studies only assessed single pre procedural levels. Previous studies assessing BNP rise following balloon angioplasty or coronary stenting have reported conflicting results; most of these studies excluded patients with recent acute coronary syndrome (ACS). RESULTS: We studied the changes in BNP following coronary stenting in 100 patients across the entire spectrum of ACS and observed a significant rise in BNP following stenting. Baseline BNP levels > 100 pg/ml were observed in 31% of patients; following PCI, 45% of patients were noted to have BNP > 100 pg/ml. In patients with baseline BNP < 100 pg/ml, 20% had post-procedure BNP levels > 100 pg/ml. Post-PCI BNP levels were significantly higher in patients with recent ACS (versus those with stable angina), those with LV dysfunction, high baseline troponin I, visible angiographic thrombus and those with post-PCI TIMI 1-2 flow. Patients with post PCI BNP levels > 100 pg/ml had a trend toward more frequent occurrence of TIMI no reflow following PCI (9.3 versus 1.7%; p = 0.07). CONCLUSION: Recent ACS, raised basal troponin, LV dysfunction, and presence of angiographic thrombus were all associated with high baseline and post-PCI BNP levels. Though recent ACS was the strongest predictor of elevated BNP levels, BNP levels rose following PCI even in patients with chronic stable angina. PMID- 21646651 TI - Subacute left ventricular rupture supported with a percutaneous left ventricular assist device. AB - Cardiac rupture is a fatal complication of transmural myocardial infarction that is associated with high mortality. We describe the successful management of a case of subacute cardiac rupture and cardiogenic shock supported by a percutaneous left ventricular assist device (LVAD) as a bridge to surgery. PMID- 21646652 TI - Multivessel intervention and placement of a percutaneous right ventricular assist device in a patient with acute myocardial infarction complicated by cardiac arrest. AB - We present a case of a 66-year-old male who presented with ST elevation myocardial infarction and complicated by cardiac arrest. The patient underwent emergent multivessel revascularization. However, the patient developed right ventricular failure with persistent and recalcitrant hemodynamic instability. Placement of a TandemHeartTM right ventricular assist deviceTM (Cardiac Assist, Pittsburgh, Pennsylvania) was undertaken to allow for right ventricular recovery. The device was removed after several days and the patient has had no further events one year after arrest. PMID- 21646653 TI - Right ventricular assist devices in right ventricular infarction: do they augment right ventricular function sufficiently to improve prognosis? PMID- 21646654 TI - Isolated single coronary artery arising from the right sinus of Valsalva. PMID- 21646655 TI - Single coronary artery anomaly: right coronary artery originating from the mid left anterior descending artery. AB - We present three cases of a single coronary artery that is the anomalous RCA originating from the mid LAD artery. These cases are rare. We discuss how to make accurate diagnosis and select appropriate treatment. PMID- 21646656 TI - Simultaneous two-vessel very late stent thrombosis and coronary aneurysm formation after sirolimus-eluting stent implantation: an intravascular ultrasound evaluation. AB - Very late stent thrombosis is a rare but devastating complication. Several factors are known to contribute to its occurrence, and are related to the patient, procedure, lesion and premature discontinuation of antiplatelet therapy. This report describes the case of a 49-year-old patient with simultaneous two vessel stent thrombosis (left anterior descending and circumflex arteries) 24 months after sirolimus-eluting stent implantation. The importance of intravascular ultrasound is enhanced. The potential contributing factors to the stent thrombosis are analyzed. PMID- 21646657 TI - Successful stenting of total left subclavian artery occlusion post-coronary artery bypass graft surgery using dual left vertebral artery and left internal mammary artery protection. AB - We report on a 68-year-old male patient (smoker, chronic obstructive pulmonary disease, diabetic for 25 years) who was admitted with acute pulmonary edema 5 months after undergoing coronary artery bypass grafting (left internal mammary artery to left anterior descending artery, saphenous vein graft to posterior descending artery, and sequential saphenous vein graft to obtuse marginal artery). He had no cardiac enzyme leakage and his left ventricular ejection fraction was > 50% on 2-dimensional echocardiography. He proved to have total calcific left subclavian artery occlusion and retrograde flow in both his left internal mammary artery and his left vertebral artery upon left subclavian angiography performed through the left brachial approach. Angiographic vertebral and coronary-subclavian steals were both supported with clinical manifestations. The patient underwent successful stenting to his left subclavian artery using dual protection to his left vertebral artery with filter protection device and to his left internal mammary artery using simple balloon inflation at its mouth before and after each subclavian artery angioplasty step. Three months later, the patient was free from any cardiac or neurologic complaints. PMID- 21646658 TI - Prasugrel as a safe alternative for clopidogrel-associated arthritis. AB - Clopidogrel is a thienopyridine derivative antiplatelet compound. The antiplatelet effects of clopidogrel originate through noncompetitive antagonism of the platelet ADP receptor, P2Y12, resulting in inhibition of platelet activation. Clopidogrel is now widely used in acute coronary syndromes and after percutaneous coronary interventions to reduce the risk of subsequent cardiovascular events. We report a case of acute migratory polyarthritis associated with the use of clopidogrel. This serves as only the second documented case of clopidogrel-associated arthritis in the United States, and the first to show that prasugrel may be considered as an alternative agent without short-term reoccurrence. PMID- 21646659 TI - Late cardiac perforation following percutaneous atrial septal defect closure using the Solysafe device. AB - Percutaneous atrial septal defect closure has been demonstrated to be a safe and effective procedure. Many different devices have been utilized during the last decade with a low complication rate. We report a patient who presented with cardiac tamponade 6 months after ASD closure using the Solysafe device. Emergency surgery demonstrated a disarticulated device as well as a broken wire migrated into the pericardial space causing cardiac perforation and tamponade. To the best of our knowledge, this is the first report of late cardiac perforation following ASD closure using the Solysafe device. PMID- 21646660 TI - A common electrocardiographic finding in an uncommon presentation. AB - Anterior ST-segment elevation is the hallmark electrocardiographic finding of acute anterior left ventricular infarction that is caused by occlusion in one of the branches of the left coronary artery. We report a case of marked ST-segment elevation in the precordial leads, with concomitant inferior ST-segment elevation that was caused by acute occlusion of the marginal branches of the right coronary artery (RCA) causing an isolated right ventricular myocardial infarction (RVMI) during coronary angioplasty. Isolated occlusion of the marginal branches of the RCA appears to be extremely rare. In the case presented, this was caused by an iatrogenic spiral dissection with subsequent stenting of the RCA. This case illustrates that diffuse ST-segment elevation in the precordial leads may occur due to the occlusion of the RCA or its branches. It is important to recognize this scenario, since the treatment of left ventricular myocardial infarction differs from that of RVMI, where maintaining adequate preload and avoiding vasodilators to preserve right ventricular stroke volume is crucial. PMID- 21646661 TI - Percutaneous transvenous mitral commissurotomy via left femoral vein approach- exploring an unusual approach for left atrial entry. AB - Percutaneous transvenous mitral commissurotomy (PTMC) using Inoue technique through a right femoral vein approach (RFVA) is well established. Left femoral vein approach (LFVA) is traditionally contraindicated because of certain technical issues. We report a case of PTMC successfully done through this unusual approach, which should be reserved exclusively for cases with right femoral vein occlusion. It also opens up the avenue for other interventions requiring a septal puncture, left atrial entry and use of bulky devices. PMID- 21646662 TI - Microcoil embolization in post-traumatic high-flow priapism. AB - An 18-year-old male developed post-traumatic priapism subsequent to a fall causing blunt perineal trauma. Color Doppler ultrasound showed a high-flow arterio-venous fistula with feeders from branches of the left internal iliac artery. Selective arteriography of the left internal pudendal artery demonstrated an arterio-venous fistula. A curved-tip 2.3 French microcatheter (Terumo Medical Corporation, Tokyo, Japan) was advanced proximal to the fistula over a 0.014" flexible guidewire. The fistula was then embolized with 2 platinum 18-2-2 Hilal Embolization Microcoils (Cook, Bloomington, Indiana). Improvement was noted, with closure of the fistula at check angiography and significant detumescence on table. This was later confirmed on repeat color Doppler imaging. At follow-up 6 and 12 weeks later, he had normal erectile function. Transarterial embolization appears to be a safe and effective treatment for managing patients with high-flow priapism. PMID- 21646663 TI - Metal fatigue in myocardial bridges: stent fracture limits the efficacy of drug eluting stents. AB - Myocardial bridging (MB) is a common anatomical varient in which a segment of coronary artery takes an intramural path. Occasionally, it can result in symptomatic ischemia. We present four cases in which MB was treated with drug eluting stents that subsequently fractured, leading to recurrent symptoms. PMID- 21646664 TI - Successful transradial intervention by switching from 6 French to 5 French guiding catheter. AB - We report a case of percutaneous coronary intervention (PCI) where a 6 French (Fr) guiding catheter could not be advanced through extremely tortuous subclavian and brachiocephalic arteries with a right transradial approach. Downsizing from a 6 Fr to 5 Fr guiding catheter was effective to reach to the coronary cusp, and we successfully performed transradial PCI without access-site crossover. When a catheter cannot be advanced in a tortuous vessel during transradial intervention, downsizing the catheter is an option to reduce resistance and enable successful PCI without access-site crossover. PMID- 21646665 TI - An unusual coronary trinity: single left coronary artery with the right coronary artery originating from the left main coronary artery and following a retro aortic course. AB - All three coronary arteries originating from a single coronary ostium is a rarity. Single coronary artery (CA) arising from the left aortic sinus is far more uncommon than one arising from the right sinus. Usually in such cases, the right coronary artery (RCA) arises as a distal continuation of the left circumflex artery. We describe an extremely uncommon anatomic variant, where the RCA arose from the single undivided CA as a branch of the left main coronary artery and followed a retro-aortic course before reaching its designated anatomic territory. PMID- 21646666 TI - Hepatitis C: many small steps. PMID- 21646667 TI - Malignancies associated with chronic hepatitis C: case report and review of the literature. AB - Hepatocellular carcinoma (HCC) is a well-known consequence of hepatitis C virus (HCV) infection mainly in cirrhotic patients. Associations of other malignancies such as cholangiocellular carcinoma and B-cell malignancies with HCV are less well known. Here we review pathophysiological aspects of malignancies associated with HCV infection. A case report of HCV-related HCC and B-cell lymphoma illustrates the increased risk for HCV-infected patients to develop other malignancies besides HCC. PMID- 21646668 TI - Treatment extension benefits HCV genotype 1 patients without rapid virological response: a systematic review. AB - BACKGROUND: Current guidelines recommend 48 weeks of treatment with pegylated interferon and ribavirin for patients infected with chronic hepatitis C virus (HCV) genotype 1. Several clinical trials have investigated the efficacy of treatment duration longer than 48 weeks, but yielded discordant results. METHODS: We performed a structured search of PubMed, Web of Science and the Cochrane library to identify randomised clinical trials in HCV genotype 1 patients who were treated either for 48 or 72 weeks. Sustained viral response (SVR) data were pooled and a sample size weighted pooled proportion was calculated. RESULTS: We identified five studies matching our criteria. Studies randomised at baseline (n=1), at absence of rapid virological response (RVR) at week 4 (n=1), at early virological response at week 12 (EVR) (n=1) or at slow response at week 24 (n=2). In the RCT that randomised at absence of RVR, SVR was significantly higher in the extended treatment arm (57 vs 42%, p=0.02) with an RR of 1.35 (95% CI 1.04 to 1.75). This tendency was also observed in the studies that randomised at slow response (44 vs 35%), although no longer statistically significantly different. CONCLUSION: Prolonged 72-week treatment should be considered in HCV genotype 1 patients without RVR at week 4, as this increased SVR. PMID- 21646669 TI - Episodes of abdominal pain. PMID- 21646670 TI - H1N1 vaccination: expect the unexpected. PMID- 21646671 TI - Illness-induced changes in thyroid hormone metabolism: focus on the tissue level. AB - During illness changes in thyroid hormone metabolism occur, collectively known as the non-thyroidal illness syndrome (NTIS). NTIS is characterised by low serum thyroid hormone levels without the expected rise in serum thyroid-stimulating hormone, indicating a major change in thyroid hormone feedback regulation. Recent studies have made clear that during NTIS differential changes in thyroid hormone metabolism occur in various tissues, the net effect of which may be either activation or inhibition of thyroid hormone action. In this review we discuss systemic and local changes in thyroid hormone metabolism during illness, highlighting their physiological implications in terms of disease course. PMID- 21646672 TI - Hospital specific factors affect quality of blood pressure treatment in chronic kidney disease. AB - BACKGROUND: Blood pressure (BP) is the most important modifiable risk factor for cardiovascular (CV) disease and progression of kidney dysfunction in patients with chronic kidney disease. Despite extensive antihypertensive treatment possibilities, adequate control is notoriously hard to achieve. Several determinants have been identified which affect BP control. In the current analysis we evaluated differences in achieved BP and achievement of the BP goal between hospitals and explored possible explanations. METHODS: At baseline, BP was measured in a supine position with an oscillometric device in 788 patients participating in the MASTER PLAN study. We also retrieved the last measured office BP from the patient records. Additional baseline characteristics were derived from the study database. Univariate and multivariate analyses were performed with general linear modelling using hospital as a random factor. RESULTS: In univariate analysis, hospital was a determinant of the level of systolic and diastolic BP at baseline. Adjustment for patient, kidney disease, treatment or hospital characteristics affected the relation. Yet, in a fully adjusted model, differences between centres persisted with a range of 15 mmHg for systolic BP and 11 mmHg for diastolic BP. CONCLUSION: Despite extensive adjustments, a clinically relevant, statistically significant difference between hospitals was found in standardised BP measurements at baseline of a randomised controlled study. We hypothesise that differences in the approach towards BP control exist at the physician level and that these explain the differences between hospitals. PMID- 21646673 TI - Episodes of shortness of breath induced by prednisone. AB - Although anecdotal reports pointing to the occurrence of episodes of shortness of breath due to prednisone use have been published, systematic evidence is lacking. In this manuscript we report on an n=1 trial in a patient using prednisone for polymyalgia rheumatica. With this approach we can confirm that prednisone may cause episodes of dyspnoea and we provide potential explanations for this side effect. PMID- 21646674 TI - A round air configuration in the lower abdomen. PMID- 21646675 TI - Dutch guideline for the management of hypertensive crisis -- 2010 revision. AB - Hypertensive crises are divided into hypertensive urgencies and emergencies. Together they form a heterogeneous group of acute hypertensive disorders depending on the presence or type of target organs involved. Despite better treatment options for hypertension, hypertensive crisis and its associated complications remain relatively common. In the Netherlands the number of patients starting renal replacement therapy because of 'malignant hypertension' has increased in the past two decades. In 2003, the first Dutch guideline on hypertensive crisis was released to allow a standardised evidence-based approach for patients presenting with a hypertensive crisis. In this paper we give an overview of the current management of hypertensive crisis and discuss several important changes incorporated in the 2010 revision. These changes include a modification in terminology replacing 'malignant hypertension' with 'hypertensive crisis with retinopathy and reclassification of hypertensive crisis with retinopathy under hypertensive emergencies instead of urgencies. With regard to the treatment of hypertensive emergencies, nicardipine instead of nitroprusside or labetalol is favoured for the management of perioperative hypertension, whereas labetalol has become the drug of choice for the treatment of hypertension associated with pre-eclampsia. For the treatment of hypertensive urgencies, oral administration of nifedipine retard instead of captopril is recommended as first line therapy. In addition, a section on the management of hypertensive emergencies according to the type of target organ involved has been added. Efforts to increase the awareness and treatment of hypertension in the population at large may lower the incidence of hypertensive crisis and its complications. PMID- 21646676 TI - Comment to case report on eosinophilic gastroenteritis. PMID- 21646677 TI - A Tensor-Based Algorithm for High-Order Graph Matching. AB - This paper addresses the problem of establishing correspondences between two sets of visual features using higher order constraints instead of the unary or pairwise ones used in classical methods. Concretely, the corresponding hypergraph matching problem is formulated as the maximization of a multilinear objective function over all permutations of the features. This function is defined by a tensor representing the affinity between feature tuples. It is maximized using a generalization of spectral techniques where a relaxed problem is first solved by a multidimensional power method and the solution is then projected onto the closest assignment matrix. The proposed approach has been implemented, and it is compared to state-of-the-art algorithms on both synthetic and real data. PMID- 21646678 TI - Scalable Face Image Retrieval with Identity-Based Quantization and Multireference Reranking. AB - State-of-the-art image retrieval systems achieve scalability by using a bag-of words representation and textual retrieval methods, but their performance degrades quickly in the face image domain, mainly because they produce visual words with low discriminative power for face images and ignore the special properties of faces. The leading features for face recognition can achieve good retrieval performance, but these features are not suitable for inverted indexing as they are high-dimensional and global and thus not scalable in either computational or storage cost. In this paper, we aim to build a scalable face image retrieval system. For this purpose, we develop a new scalable face representation using both local and global features. In the indexing stage, we exploit special properties of faces to design new component-based local features, which are subsequently quantized into visual words using a novel identity-based quantization scheme. We also use a very small Hamming signature (40 bytes) to encode the discriminative global feature for each face. In the retrieval stage, candidate images are first retrieved from the inverted index of visual words. We then use a new multireference distance to rerank the candidate images using the Hamming signature. On a one millon face database, we show that our local features and global Hamming signatures are complementary--the inverted index based on local features provides candidate images with good recall, while the multireference reranking with global Hamming signature leads to good precision. As a result, our system is not only scalable but also outperforms the linear scan retrieval system using the state-of-the-art face recognition feature in term of the quality. PMID- 21646679 TI - Domain transfer multiple kernel learning. AB - Cross-domain learning methods have shown promising results by leveraging labeled patterns from the auxiliary domain to learn a robust classifier for the target domain which has only a limited number of labeled samples. To cope with the considerable change between feature distributions of different domains, we propose a new cross-domain kernel learning framework into which many existing kernel methods can be readily incorporated. Our framework, referred to as Domain Transfer Multiple Kernel Learning (DTMKL), simultaneously learns a kernel function and a robust classifier by minimizing both the structural risk functional and the distribution mismatch between the labeled and unlabeled samples from the auxiliary and target domains. Under the DTMKL framework, we also propose two novel methods by using SVM and prelearned classifiers, respectively. Comprehensive experiments on three domain adaptation data sets (i.e., TRECVID, 20 Newsgroups, and email spam data sets) demonstrate that DTMKL-based methods outperform existing cross-domain learning and multiple kernel learning methods. PMID- 21646680 TI - Toward a practical face recognition system: robust alignment and illumination by sparse representation. AB - Many classic and contemporary face recognition algorithms work well on public data sets, but degrade sharply when they are used in a real recognition system. This is mostly due to the difficulty of simultaneously handling variations in illumination, image misalignment, and occlusion in the test image. We consider a scenario where the training images are well controlled and test images are only loosely controlled. We propose a conceptually simple face recognition system that achieves a high degree of robustness and stability to illumination variation, image misalignment, and partial occlusion. The system uses tools from sparse representation to align a test face image to a set of frontal training images. The region of attraction of our alignment algorithm is computed empirically for public face data sets such as Multi-PIE. We demonstrate how to capture a set of training images with enough illumination variation that they span test images taken under uncontrolled illumination. In order to evaluate how our algorithms work under practical testing conditions, we have implemented a complete face recognition system, including a projector-based training acquisition system. Our system can efficiently and effectively recognize faces under a variety of realistic conditions, using only frontal images under the proposed illuminations as training. PMID- 21646681 TI - A novel word spotting method based on recurrent neural networks. AB - Keyword spotting refers to the process of retrieving all instances of a given keyword from a document. In the present paper, a novel keyword spotting method for handwritten documents is described. It is derived from a neural network-based system for unconstrained handwriting recognition. As such it performs template free spotting, i.e., it is not necessary for a keyword to appear in the training set. The keyword spotting is done using a modification of the CTC Token Passing algorithm in conjunction with a recurrent neural network. We demonstrate that the proposed systems outperform not only a classical dynamic time warping-based approach but also a modern keyword spotting system, based on hidden Markov models. Furthermore, we analyze the performance of the underlying neural networks when using them in a recognition task followed by keyword spotting on the produced transcription. We point out the advantages of keyword spotting when compared to classic text line recognition. PMID- 21646682 TI - Retraction: Towards prediction and prioritization of disease genes by the modularity of human phenome-genome assembled network. PMID- 21646683 TI - Myeloproliferative neoplasms: from JAK2 mutations discovery to JAK2 inhibitor therapies. AB - Most BCR-ABL1-negative myeloproliferative neoplasms (MPN) carry an activating JAK2 mutation. Approximately 96% of patients with polycythemia vera (PV) harbors the V617F mutation in JAK2 exon 14, whereas the minority of JAK2 (V617F)-negative subjects shows several mutations in exon 12. Other mutation events as MPL, TET2, LNK, EZH2 have been described in chronic phase, while NF1, IDH1, IDH2, ASX1, CBL and Ikaros in blast phase of MPN. The specific pathogenic implication of these mutations is under investigation, but they may have a role in refinement of diagnostic criteria and in development of new prognostic models. Several trials with targeted therapy (JAK inhibitors) are ongoing mostly involving patients with PMF, post-PV MF and post-essential thrombocythemia (ET) MF. Treatment with ruxolitinib and TG101348 has shown clinically significant benefits, particularly in improvement of splenomegaly and constitutional symptoms in MF patients. On the other hand, JAK inhibitors have not thus far shown disease-modifying activity therefore any other deduction on these new drugs seems premature. PMID- 21646684 TI - Molecular signature induced by RNASET2, a tumor antagonizing gene, in ovarian cancer cells. AB - Using the Hey3Met2 human ovarian cancer cell line, we previously found the RNASET2 gene to possess a remarkable in vivo tumor suppressor activity, although no in vitro features such as inhibition of cell proliferation, clonogenic potential, impaired growth in soft agar and increase in apoptotic rate could be detected. This is reminiscent of the behavior of genes belonging to the class of tumor antagonizing genes (TAG) which act mainly within the context of the microenvironment. Here we present transcriptional profiles analysis which indicates that investigations of the mechanisms of TAG biological functions require a comparison between the in vitro and in vivo expression patterns. Indeed several genes displaying a biological function potentially related to tumor suppression could not be validated by subsequent in vivo expression analysis. On the other hand the fact that we could find congruency for three genes both in vivo and in vitro adds a warning to a too much stringent categorization of this class of genes which relies on the sensitivity of the methodological approaches. PMID- 21646686 TI - Strong inhibition of xenografted tumor growth by low-level doses of [(32)P]ATP. AB - The ability of a potential human anti-cancer therapeutic agent to inhibit the growth of xenografted tumors in nude mice has been an established and accepted testing method for several decades. The current report shows that a single, low level intravenous dose of [(32)P]ATP significantly inhibits the growth of established xenografted tumors in nude mice. This inhibitory effect becomes appreciable very rapidly, within only five days post-injection and the low dose demonstrates little or no toxicity in the mice. Surprisingly, a narrow dose window of optimum effectiveness is seen, whereby either decreasing or increasing the [(32)P]ATP dose results in far less growth inhibition. Thus, the intravenous systemic injection of [(32)P]ATP may represent a simple, potent method to target and inhibit primary human tumors and malignant lesions. PMID- 21646685 TI - Elevated PI3K signaling drives multiple breast cancer subtypes. AB - Most human breast tumors have mutations that elevate signaling through a key metabolic pathway that is induced by insulin and a number of growth factors. This pathway serves to activate an enzyme known as phosphatidylinositol 3' kinase (PI3K) as well as to regulate proteins that signal in response to lipid products of PI3K. The specific mutations that activate this pathway in breast cancer can occur in genes coding for tyrosine kinase receptors, adaptor proteins linked to PI3K, catalytic and regulatory subunits of PI3K, serine/threonine kinases that function downstream of PI3K, and also phosphatidylinositol 3' phosphatase tumor suppressors that function to antagonize this pathway. While each genetic change results in net elevation of PI3K pathway signaling, and all major breast cancer subtypes show pathway activation, the specific mutation(s) involved in any one tumor may play an important role in defining tumor subtype, prognosis and even sensitivity to therapy. Here, we describe mouse models of breast cancer with elevated PI3K signaling, and how they may be used to guide development of novel therapeutics. PMID- 21646688 TI - Fractional exhaled nitric oxide measurement with a handheld device. AB - A sensing system for fractional exhaled nitric oxide (FeNO) measurement is presented, which is characterized by a compact setup and a cost potential to be made available for the patient at home. The sensing is based on the work function measurement of a phthalocyanine-type sensing material, which is shown to be sufficiently sensitive for NO(2) in the ppb range. The transducer used to measure the work function is a field effect transistor with a suspended gate electrode. Selectivity is given with respect to other breath components including typically metabolic by-products. The measurement system includes breath treatments in a simple setup, which essentially are dehumidification and a quantitative conversion of NO to NO(2) with a conversion rate of approx. 95%, using a disposable oxidation catalyst. The accomplishment of the correct exhalation maneuver and feeding of the suited portion of exhaled air to the sensor is provided by breath sampling means. The sensor is not gas consuming. This allows us to fill the measurement chamber once, instead of establishing a gas flow for the measurement. This feature simplifies the device architecture. In this paper, we report on sensor characteristics, system architecture and measurement with artificial breath-gas as well as with human breath with the device. PMID- 21646687 TI - The direct Myc target Pim3 cooperates with other Pim kinases in supporting viability of Myc-induced B-cell lymphomas. AB - The Pim kinases are weak oncogenes. However, when co-expressed with a strong oncogene, such as c-Myc, Pim kinases potentiate the oncogenic effect resulting in an acceleration of tumorigenesis. In this study we show that the least studied Pim kinase, Pim-3, is encoded by a gene directly regulated by c-Myc via binding to one of the conserved E-boxes within the Pim3 gene. Accordingly, lymphomas arising in Myc-transgenic mice and Burkitt lymphoma cell lines exhibit elevated levels of Pim-3. Interestingly, inhibition of Pim kinases by a novel pan-Pim kinase inhibitor, Pimi, in Myc-induced lymphoma results in cell death that appears independent of caspases. The data indicate that Pim kinase inhibition could be a viable treatment strategy in certain human lymphomas that rely on Pim 3 kinase expression. PMID- 21646689 TI - Background and survey of bioreplication techniques. AB - Bioreplication is the direct reproduction of a biological structure in order to realize at least one specific functionality. Current bioreplication techniques include the sol-gel technique, atomic layer deposition, physical vapor deposition, and imprint lithography and casting. The combined use of a focused ion beam and a scanning electron microscope could develop into a bioreplication technique as well. Some of these techniques are more suitable for reproducing surface features, others for bulk three-dimensional structures. Industrial upscaling appears possible only for imprint lithography and casting (which can be replaced by stamping). PMID- 21646690 TI - Cathodoluminescence of rare earth implanted Ga2O3 and GeO2 nanostructures. AB - Rare earth (RE) doped gallium oxide and germanium oxide micro- and nanostructures, mostly nanowires, have been obtained and their morphological and optical properties have been characterized. Undoped oxide micro- and nanostructures were grown by a thermal evaporation method and were subsequently doped with gadolinium or europium ions by ion implantation. No significant changes in the morphologies of the nanostructures were observed after ion implantation and thermal annealing. The luminescence emission properties have been studied with cathodoluminescence (CL) in a scanning electron microscope (SEM). Both beta-Ga(2)O(3) and GeO(2) structures implanted with Eu show the characteristic red luminescence peak centered at around 610 nm, due to the (5)D(0)-(7)F(2) Eu(3+) intraionic transition. Sharpening of the luminescence peaks after thermal annealing is observed in Eu implanted beta-Ga(2)O(3), which is assigned to the lattice recovery. Gd(3+) as-implanted samples do not show rare earth related luminescence. After annealing, optical activation of Gd(3+) is obtained in both matrices and a sharp ultraviolet peak centered at around 315 nm, associated with the Gd(3+) (6)P(7/2)-(8)S(7/2) intraionic transition, is observed. The influence of the Gd ion implantation and the annealing temperature on the gallium oxide broad intrinsic defect band has been analyzed. PMID- 21646691 TI - Temperature-dependent evolution of the wetting layer thickness during Ge deposition on Si(001). AB - The evolution of the wetting layer (WL) thickness during Ge deposition on Si(001) is analyzed with the help of a rate-equation approach. The combined role of thickness, island volume and shape-dependent chemical potentials is considered. Several experimental observations, such as WL thinning following the pyramid-to dome transformation, are captured by the model, as directly demonstrated by a close comparison with photoluminescence measurements (PL) on samples grown at three different temperatures. The limitations of the model in describing late stages of growth are critically addressed. PMID- 21646692 TI - A novel hybrid organic/inorganic photonic crystal slab showing a resonance action at the band edge. AB - Here we propose and experimentally demonstrate a hybrid photonic crystal (PC) slab consisting of air rods in a nanocomposite prepared by incorporating CdSe/CdS core/shell NRs (NR) in a polymer. Since the styrene methyl acrylate based polymer (ZEP) is transparent in the visible spectral range and is an electron-sensitive material, it was chosen as the embedding matrix for the NRs. Scanning electron microscopy and luminance measurements were used to characterize the experimental structure. The vertical extraction of the light, by the coupling of the modes guided by the PC slab to the free radiation via Bragg scattering, consists of a narrow orange emission band at 592 nm with a full width at half-maximum (FWHM) of 17 nm. The original characteristics of hybrid materials based on polymers and colloidal NRs, able to combine the unique optical properties of the inorganic moiety with the processability of the host matrix, are extremely appealing in view of their technological impact on the development of new high performing optical devices such as organic light-emitting diodes, ultra-low threshold lasers and non-linear devices. PMID- 21646693 TI - Surface imaging using holographic optical tweezers. AB - We present an imaging technique using an optically trapped cigar-shaped probe controlled using holographic optical tweezers. The probe is raster scanned over a surface, allowing an image to be taken in a manner analogous to scanning probe microscopy (SPM), with automatic closed loop feedback control provided by analysis of the probe position recorded using a high speed CMOS camera. The probe is held using two optical traps centred at least 10 um from the ends, minimizing laser illumination of the tip, so reducing the chance of optical damage to delicate samples. The technique imparts less force on samples than contact SPM techniques, and allows highly curved and strongly scattering samples to be imaged, which present difficulties for imaging using photonic force microscopy. To calibrate our technique, we first image a known sample--the interface between two 8 um polystyrene beads. We then demonstrate the advantages of this technique by imaging the surface of the soft alga Pseudopediastrum. The scattering force of our laser applied directly onto this sample is enough to remove it from the surface, but we can use our technique to image the algal surface with minimal disruption while it is alive, not adhered and in physiological conditions. The resolution is currently equivalent to confocal microscopy, but as our technique is not diffraction limited, there is scope for significant improvement by reducing the tip diameter and limiting the thermal motion of the probe. PMID- 21646694 TI - Numerical study of the lateral resolution in electrostatic force microscopy for dielectric samples. AB - We present a study of the lateral resolution in electrostatic force microscopy for dielectric samples in both force and gradient modes. Whereas previous studies have reported expressions for metallic surfaces having potential heterogeneities (Kelvin probe force microscopy), in this work we take into account the presence of a dielectric medium. We introduce a definition of the lateral resolution based on the force due to a test particle being either a point charge or a polarizable particle on the dielectric surface. The behaviour has been studied over a wide range of typical experimental parameters: tip-sample distance (1-20) nm, sample thickness (0-5) um and dielectric constant (1-20), using the numerical simulation of the equivalent charge method. For potential heterogeneities on metallic surfaces expressions are in agreement with the bibliography. The lateral resolution of samples having a dielectric constant of more than 10 tends to metallic behaviour. We found a characteristic thickness of 100 nm, above which the lateral resolution measured on the dielectric surface is close to that of an infinite medium. As previously reported, the lateral resolution is better in the gradient mode than in the force mode. Finally, we showed that for the same experimental conditions, the lateral resolution is better for a polarizable particle than for a charge, i.e. dielectric heterogeneities should always look 'sharper' (better resolved) than inhomogeneous charge distributions. This fact should be taken into account when interpreting images of heterogeneous samples. PMID- 21646695 TI - Vanishing stick-slip friction in few-layer graphenes: the thickness effect. AB - We report the thickness dependence of intrinsic friction in few-layer graphenes, adopting molecular dynamics simulations. The friction force drops dramatically with decreasing number of layers and finally approaches zero with two or three layers. The results, which are robust over a wide range of temperature, shear velocity, and pressure are quantitatively explained by a theoretical model with regard to lateral stiffness, slip length, and maximum lateral force, which could provide a new conceptual framework for understanding stick-slip friction. The results reveal the crucial role of the dimensional effect in nanoscale friction, and could be helpful in the design of graphene-based nanodevices. PMID- 21646696 TI - Large-area regular nanodomain patterning in He-irradiated lithium niobate crystals. AB - Large-area ferroelectric nanodomain patterns, which are desirable for nonlinear optical applications, were generated in previously He-implanted lithium niobate crystals by applying voltage pulses to the tip of a scanning force microscope. The individual nanodomains were found to be of uniform size, which depended only on the inter-domain spacing and the pulse amplitude. We explain this behavior by the electrostatic repulsion of poling-induced buried charges between adjacent domains. The domain patterns were imaged by piezoresponse force microscopy and investigated by domain-selective etching in conjunction with focused ion beam etching followed by scanning electron microscopy imaging. In order to optimize the He-irradiation parameters for easy and reliable nanodomain patterning a series of samples subjected to various irradiation fluences and energies was prepared. The different samples were characterized by investigating nanodomains generated with a wide range of pulse parameters (amplitude and duration). In addition, these experiments clarified the physical mechanism behind the facile poling measured in He-irradiated lithium niobate crystals: the damage caused by the energy loss that takes place via electronic excitations appears to act to stabilize the domains, whereas the nuclear-collision damage degrades the crystal quality, and thus impedes reliable nanodomain generation. PMID- 21646697 TI - Raman scattering and growth disorders in single as-grown TiO2 nanowires. AB - An oxidation procedure has been developed to grow single-crystalline TiO(2) nanowires of the pure rutile phase, allowing subsequent characterizations of SEM, XRD, Raman, and TEM without any post-growth preparations. TEM observations support that the 1D anisotropic growth is dominated by oriented attachment processes, leading to typical growth-induced defects in the nanowires. Spatial variations of the rutile E(g) and A(1g) Raman modes were unambiguously revealed on single nanowires while scanned along the growth direction parallel to the rutile [110]. Symmetry-sensitive deviations were identified by comparing the Raman data with the spatial correlation model calculations based on realistic dispersion relations of the rutile, reflecting morphology-correlated defect distributions along single nanowires. This work provides an efficient, non destructive in situ characterization approach for guiding growth design in future nanotechnology. PMID- 21646698 TI - Manipulation of individual double-walled carbon nanotubes packed in a casing shell. AB - Controlled placement of carbon nanotubes is important for carbon-based nanodevice assembly. However, it is difficult to manipulate individual nanotubes because of their extremely small dimensions. Ultra-fine tubes are often in the form of bundles and are hard to efficiently move on a surface due to the strong adhesion among themselves and between the tubes and the substrate. This paper presents a novel manipulation approach of individual double-walled carbon nanotubes encased in a thick amorphous carbon shell. With an atomic force microscope, we are able to freely displace the nanotubes within a casing shell, and unpack it from the shell on a silicon surface. The theoretical analysis demonstrates that the unpacking process is determined by the difference of the static friction between the shell and the substrate and the resistance force between the shell and the embedded nanotube. PMID- 21646699 TI - Data-driven classification of ventilated lung tissues using electrical impedance tomography. AB - Current methods for identifying ventilated lung regions utilizing electrical impedance tomography images rely on dividing the image into arbitrary regions of interest (ROI), manually delineating ROI, or forming ROI with pixels whose signal properties surpass an arbitrary threshold. In this paper, we propose a novel application of a data-driven classification method to identify ventilated lung ROI based on forming k clusters from pixels with correlated signals. A standard first-order model for lung mechanics is then applied to determine which ROI correspond to ventilated lung tissue. We applied the method in an experimental study of 16 mechanically ventilated swine in the supine position, which underwent changes in positive end-expiratory pressure (PEEP) and fraction of inspired oxygen (F(I)O(2)). In each stage of the experimental protocol, the method performed best with k = 4 and consistently identified 3 lung tissue ROI and 1 boundary tissue ROI in 15 of the 16 subjects. When testing for changes from baseline in lung position, tidal volume, and respiratory system compliance, we found that PEEP displaced the ventilated lung region dorsally by 2 cm, decreased tidal volume by 1.3%, and increased the respiratory system compliance time constant by 0.3 s. F(I)O(2) decreased tidal volume by 0.7%. All effects were tested at p < 0.05 with n = 16. These findings suggest that the proposed ROI detection method is robust and sensitive to ventilation dynamics in the experimental setting. PMID- 21646700 TI - The feasibility of transoesophageal bioimpedance measurements for the detection of heart graft rejection. AB - Previous studies demonstrate that it is possible to evaluate a heart graft rejection condition using a bioimpedance technique by means of an intracavitary catheter. We propose to use a less invasive technique consisting in the use of a transoesophageal catheter and two standard ECG electrodes on the thorax. The aim of this work is to evaluate, using the finite element method, several parameters affecting the transoesophageal impedance measurement, including sensitivity to electrical conductivity and permittivity of different organs in the thorax, changes in magnitude and phase due to a lesion producing a scar, a global ischaemia of the heart, pleural effusion in the lungs, fat thickness increase, displacement of the catheter inside the oesophagus and movement of one electrode on the thorax surface. From these results, we deduce the best estimator for cardiac rejection detection and obtain the tools to identify eventual cases of false positives due to other factors. To achieve these objectives we have created a thoracic model and we have simulated different situations at the frequencies of 13, 30, 100, 300 and 1000 kHz. Our simulation demonstrates that the phase, at 100 and 300 kHz, would be a better estimator than the magnitude to evaluate a heart rejection condition. PMID- 21646701 TI - A highly phase-stable differential detector amplifier for magnetic induction tomography. AB - Magnetic induction tomography (MIT) has been proposed for the detection of cerebral oedema and haemorrhagic stroke. Achieving the required phase measurement precision for these applications is however a major technical challenge. A critical component within an MIT system is the detector amplifier and for this role an ultra-phase-stable, low noise instrumentation amplifier has been developed. The design of the amplifier is described and (i) the results of simulations and measurements of the amplifiers phase stability versus temperature and (ii) measurements of the phase noise and drift performance of the amplifier within a single-channel magnetic induction spectroscopy system are provided and discussed. For a 10 MHz signal the amplifier, with a gain of 21, displayed an average change in the measured phase of its output of just -0.1 +/- 0.6 m degrees degrees C(-1) as the ambient temperature was varied between 35 and 50 degrees C, demonstrating a level of phase stability approaching that required for potential biomedical applications such as the detection of cerebral haemorrhage. PMID- 21646702 TI - Quantification of intraventricular hemorrhage with electrical impedance tomography using a spherical model. AB - We have developed a robust EEG-based current pattern which shows promise for the detection of intraventricular hemorrhage (IVH) in neonates. Our reconstructions to date are based on a layered spherical head model. In this study, the current pattern was used to gather data from three realistic-shaped neonatal head models and a physical phantom based on one of these models. We found that a sensitivity matrix calculated from a spherical model gave us satisfactory reconstructions in terms of both image quality and quantification. Incorporating correct geometry information into the forward model improved image quality. However, it did not improve quantification accuracy. The results indicate that using a spherical matrix may be a more practical choice for monitoring IVH volumes in neonates for whom patient-specific models are not available. PMID- 21646703 TI - Broadband spectroscopy of dynamic impedances with short chirp pulses. AB - An impedance spectrum of dynamic systems is time dependent. Fast impedance changes take place, for example, in high throughput microfluidic devices and in operating cardiovascular systems. Measurements must be as short as possible to avoid significant impedance changes during the spectrum analysis, and as long as possible for enlarging the excitation energy and obtaining a better signal-to noise ratio (SNR). The authors propose to use specific short chirp pulses for excitation. Thanks to the specific properties of the chirp function, it is possible to meet the needs for a spectrum bandwidth, measurement time and SNR so that the most accurate impedance spectrogram can be obtained. The chirp wave excitation can include thousands of cycles when the impedance changes slowly, but in the case of very high speed changes it can be shorter than a single cycle, preserving the same excitation bandwidth. For example, a 100 kHz bandwidth can be covered by the chirp pulse with durations from 10 us to 1 s; only its excitation energy differs also 10(5) times. After discussing theoretical short chirp properties in detail, the authors show how to generate short chirps in the microsecond range with a bandwidth up to a few MHz by using digital synthesis architectures developed inside a low-cost standard field programmable gate array. PMID- 21646704 TI - Impedance sensor technology for cell-based assays in the framework of a high content screening system. AB - Living cultured cells react to external influences, such as pharmaceutical agents, in an intricate manner due to their complex internal signal processing. Impedance sensing of cells on microelectrodes is a favored label-free technology to indicate cellular events, usually ascribed to morphologic alteration or changes in cellular adhesion, which is usually found in stand-alone systems that do not incorporate life support or additional sensor systems. However, only in symbiosis with metabolic activity sensing and picture documentation may a complete insight into cellular vitality be provided. This complement was created within the framework of an automated high-content screening system previously developed by our group, monitoring 24 cell culture chambers in parallel. The objective of this paper is the development of miniaturized electronics for impedance measurements and its system integration as a modular unit. In addition, it is shown how sensor electrodes were optimized by impedance matching such that spectroscopy and raw data analysis become feasible for every culture well. Undesired mechanical stress on cultured cells may arise from the medium and agent support system of the autonomous screening apparatus. This paper demonstrates how this hazard is treated with the simulation of microfluidics and impedance measurements. Physiological data are subsequently derived from the exemplary tumor cell line MCF-7 both during treatment with the agent doxorubicin and through the impact of natural killer cells. This correlates the information content of complex impedance spectra with cellular respiration as well as data from microscopy. PMID- 21646705 TI - Estimation of normal hydration in dialysis patients using whole body and calf bioimpedance analysis. AB - Prescription of an appropriate dialysis target weight (dry weight) requires accurate evaluation of the degree of hydration. The aim of this study was to investigate whether a state of normal hydration (DW(cBIS)) as defined by calf bioimpedance spectroscopy (cBIS) and conventional whole body bioimpedance spectroscopy (wBIS) could be characterized in hemodialysis (HD) patients and normal subjects (NS). wBIS and cBIS were performed in 62 NS (33 m/29 f) and 30 HD patients (16 m/14 f) pre- and post-dialysis treatments to measure extracellular resistance and fluid volume (ECV) by the whole body and calf bioimpedance methods. Normalized calf resistivity (rho(N)(,5)) was defined as resistivity at 5 kHz divided by the body mass index. The ratio of wECV to total body water (wECV/TBW) was calculated. Measurements were made at baseline (BL) and at DW(cBIS) following the progressive reduction of post-HD weight over successive dialysis treatments until the curve of calf extracellular resistance is flattened (stabilization) and the rho(N)(,5) was in the range of NS. Blood pressures were measured pre- and post-HD treatment. rho(N)(,5) in males and females differed significantly in NS. In patients, rho(N)(,5) notably increased with progressive decrease in body weight, and systolic blood pressure significantly decreased pre- and post-HD between BL and DW(cBIS) respectively. Although wECV/TBW decreased between BL and DW(cBIS), the percentage of change in wECV/TBW was significantly less than that in rho(N)(,5) (-5.21 +/- 3.2% versus 28 +/- 27%, p < 0.001). This establishes the use of rho(N)(,5) as a new comparator allowing a clinician to incrementally monitor removal of extracellular fluid from patients over the course of dialysis treatments. The conventional whole body technique using wECV/TBW was less sensitive than the use of rho(N)(,5) to measure differences in body hydration between BL and DW(cBIS). PMID- 21646706 TI - A fully parallel multi-frequency EIT system with flexible electrode configuration: KHU Mark2. AB - We report the development of a new multi-frequency electrical impedance tomography (EIT) system called the KHU Mark2. It is descended from the KHU Mark1 in terms of technical details such as digital waveform generation, Howland current source with multiple generalized impedance converters and digital phase sensitive demodulators. New features include flexible electrode configurations to accommodate application-specific requirements, multiple independent current sources and voltmeters for fully parallel operations, improved data acquisition speeds for faster frame rates and compact mechanical design. Given an electrode configuration, we can design an analog backplane in such a way that both current injections and voltage measurements can be done without using any switch. The KHU Mark2 is based on an impedance measurement module (IMM) comprising a current source and a voltmeter. Using multiple IMMs, we can construct a multi-channel system with 16, 32 or 64 channels, for example. Adopting a pipeline structure, it has the maximum data acquisition speed of 100 scans s(-1) with the potential to detect fast physiological changes during respiration and cardiac activity. Measuring both in-phase and quadrature components of trans-impedances at multiple frequencies simultaneously, the KHU Mark2 is apt at spectroscopic EIT imaging. In this paper, we describe its design, construction, calibration and performance evaluation. It has about 84 dB signal-to-noise ratio and 0.5% reciprocity error. Time-difference images of an admittivity phantom are presented showing spectroscopic admittivity images. Future application studies using the KHU Mark2 are briefly discussed. PMID- 21646707 TI - Evaluation of algorithms for calculating bioimpedance phase angle values from measured whole-body impedance modulus. AB - This paper addresses the problem of calculating the bioimpedance phase angle from measurements of impedance modulus. A complete impedance measurement was performed on altogether 20 healthy persons using a Solatron 1260/1294 system. The obtained impedance modulus (absolute impedance value) values were used to calculate the Cole parameters and from them the phase angles. In addition, the phase angles were also calculated using a Kramers-Kronig approach. A correlation analysis for all subjects at each frequency (5, 50, 100 and 200 kHz) for both methods gave R(2) values ranging from 0.7 to 0.96 for the Cole approach and from 0.83 to 0.96 for the Kramers-Kronig approach; thus, both methods gave good results compared with the complete measurement results. From further statistical significance testing of the absolute value of the difference between measured and calculated phase angles, it was found that the Cole equation method gave significantly better agreement for the 50 and 100 kHz frequencies. In addition, the Cole equation method gives the four Cole parameters (R(0), R(infinity), tau(z) and alpha) using measurements at frequencies up to 200 kHz while the Kramers-Kronig method used frequencies up to 500 kHz to reduce the effect of truncation on the calculated results. Both methods gave results that can be used for further bioimpedance calculations, thus improving the application potential of bioimpedance measurement results obtained using relatively inexpensive and portable measurement equipment. PMID- 21646708 TI - An oppositional biogeography-based optimization technique to reconstruct organ boundaries in the human thorax using electrical impedance tomography. AB - Electrical impedance tomography (EIT) is a non-invasive imaging modality which has been actively studied for its industrial as well as medical applications. However, the performance of the inverse algorithms to reconstruct the conductivity images using EIT is often sub-optimal. Several factors contribute to this poor performance, including high sensitivity of EIT to the measurement noise, the rounding-off errors, the inherent ill-posed nature of the problem and the convergence to a local minimum instead of the global minimum. Moreover, the performance of many of these inverse algorithms heavily relies on the selection of initial guess as well as the accurate calculation of a gradient matrix. Considering these facts, the need for an efficient optimization algorithm to reach the correct solution cannot be overstated. This paper presents an oppositional biogeography-based optimization (OBBO) algorithm to estimate the shape, size and location of organ boundaries in a human thorax using 2D EIT. The organ boundaries are expressed as coefficients of truncated Fourier series, while the conductivities of the tissues inside the thorax region are assumed to be known a priori. The proposed method is tested with the use of a realistic chest shaped mesh structure. The robustness of the algorithm has been verified, first through repetitive numerical simulations by adding randomly generated measurement noise to the simulated voltage data, and then with the help of an experimental setup resembling the human chest. An extensive statistical analysis of the estimated parameters using OBBO and its comparison with the traditional modified Newton-Raphson (mNR) method are presented. The results demonstrate that OBBO has significantly better estimation performance compared to mNR. Furthermore, it has been found that OBBO is robust to the initial guess of the size and location of the boundaries as well as offering a reasonable solution when the a priori knowledge of the conductivity of the organs is not very accurate. PMID- 21646709 TI - Adjacent stimulation and measurement patterns considered harmful. AB - We characterize the ability of electrical impedance tomography (EIT) to distinguish changes in internal conductivity distributions, and analyze it as a function of stimulation and measurement patterns. A distinguishability measure, z, is proposed which is related to the signal-to-noise ratio of a medium and to the probability of detection of conductivity changes in a region of interest. z is a function of the number of electrodes, the EIT stimulation and measurement protocol, the stimulation amplitude, the measurement noise, and the size and location of the contrasts. Using this measure we analyze various choices of stimulation and measurement patterns under the constraint of medical electrical safety limits (maximum current into the body). Analysis is performed for a planar placement of 16 electrodes for simulated 3D tank and chest shapes, and measurements in a saline tank. Results show that the traditional (and still most common) adjacent stimulation and measurement patterns have by far the poorest performance (by 6.9 *). Good results are obtained for trigonometric patterns and for pair drive and measurement patterns separated by over 90 degrees . Since the possible improvement over adjacent patterns is so large, we present this result as a call to action: adjacent patterns are harmful, and should be abandoned. We recommend using pair drive and measurement patterns separated by one electrode less than 180 degrees . We describe an approach to modify an adjacent pattern EIT system by adjusting electrode placement. PMID- 21646710 TI - The impact of electrode area, contact impedance and boundary shape on EIT images. AB - Electrical impedance tomography (EIT) measures the conductivity distribution within an object based on the current applied and voltage measured at surface electrodes. Thus, EIT images are sensitive to electrode properties (i.e. contact impedance, electrode area and boundary shape under the electrode). While some of these electrode properties have been investigated individually, this paper investigates these properties and their interaction using finite element method simulations and the complete electrode model (CEM). The effect of conformal deformations on image reconstruction when using the CEM was of specific interest. Observed artefacts were quantified using a measure that compared an ideal image to the reconstructed image, in this case a no-noise reconstruction that isolated the electrodes' effects. For electrode contact impedance and electrode area, uniform reductions to all electrodes resulted in ringing artefacts in the reconstructed images when the CEM was used, while parameter variations that were not correlated amongst electrodes resulted in artefacts distributed throughout the image. When the boundary shape changed under the electrode, as with non symmetric conformal deformations, using the CEM resulted in structured distortions within the reconstructed image. Mean electrode contact impedance increases, independent of inter-electrode variation, did not result in artefacts in the reconstructed image. PMID- 21646711 TI - Optical breast shape capture and finite-element mesh generation for electrical impedance tomography. AB - X-ray mammography is the standard for breast cancer screening. The development of alternative imaging modalities is desirable because mammograms expose patients to ionizing radiation. Electrical impedance tomography (EIT) may be used to determine tissue conductivity, a property which is an indicator of cancer presence. EIT is also a low-cost imaging solution and does not involve ionizing radiation. In breast EIT, impedance measurements are made using electrodes placed on the surface of the patient's breast. The complex conductivity of the volume of the breast is estimated by a reconstruction algorithm. EIT reconstruction is a severely ill-posed inverse problem. As a result, noisy instrumentation and incorrect modelling of the electrodes and domain shape produce significant image artefacts. In this paper, we propose a method that has the potential to reduce these errors by accurately modelling the patient breast shape. A 3D hand-held optical scanner is used to acquire the breast geometry and electrode positions. We develop methods for processing the data from the scanner and producing volume meshes accurately matching the breast surface and electrode locations, which can be used for image reconstruction. We demonstrate this method for a plaster breast phantom and a human subject. Using this approach will allow patient-specific finite-element meshes to be generated which has the potential to improve the clinical value of EIT for breast cancer diagnosis. PMID- 21646712 TI - Minimizing EIT image artefacts from mesh variability in finite element models. AB - Electrical impedance tomography (EIT) solves an inverse problem to estimate the conductivity distribution within a body from electrical simulation and measurements at the body surface, where the inverse problem is based on a solution of Laplace's equation in the body. Most commonly, a finite element model (FEM) is used, largely because of its ability to describe irregular body shapes. In this paper, we show that simulated variations in the positions of internal nodes within a FEM can result in serious image artefacts in the reconstructed images. Such variations occur when designing FEM meshes to conform to conductivity targets, but the effects may also be seen in other applications of absolute and difference EIT. We explore the hypothesis that these artefacts result from changes in the projection of the anisotropic conductivity tensor onto the FEM system matrix, which introduces anisotropic components into the simulated voltages, which cannot be reconstructed onto an isotropic image, and appear as artefacts. The magnitude of the anisotropic effect is analysed for a small regular FEM, and shown to be proportional to the relative node movement as a fraction of element size. In order to address this problem, we show that it is possible to incorporate a FEM node movement component into the formulation of the inverse problem. These results suggest that it is important to consider artefacts due to FEM mesh geometry in EIT image reconstruction. PMID- 21646713 TI - Evaluation of EIT system performance. AB - An electrical impedance tomography (EIT) system images internal conductivity from surface electrical stimulation and measurement. Such systems necessarily comprise multiple design choices from cables and hardware design to calibration and image reconstruction. In order to compare EIT systems and study the consequences of changes in system performance, this paper describes a systematic approach to evaluate the performance of the EIT systems. The system to be tested is connected to a saline phantom in which calibrated contrasting test objects are systematically positioned using a position controller. A set of evaluation parameters are proposed which characterize (i) data and image noise, (ii) data accuracy, (iii) detectability of single contrasts and distinguishability of multiple contrasts, and (iv) accuracy of reconstructed image (amplitude, resolution, position and ringing). Using this approach, we evaluate three different EIT systems and illustrate the use of these tools to evaluate and compare performance. In order to facilitate the use of this approach, all details of the phantom, test objects and position controller design are made publicly available including the source code of the evaluation and reporting software. PMID- 21646714 TI - Measurement of regional pulmonary oxygen uptake--a novel approach using electrical impedance tomography. AB - Pulmonary oxygen (O(2)) uptake during apnoea results in a fall in lung volume. Given that electrical impedance tomography (EIT) provides reliable data on regional lung volume changes we hypothesized that EIT could be used to measure regional O(2) uptake. A total of 12 lung healthy supine patients were studied. EIT measurements were performed during volume-controlled mechanical ventilation followed by apnoea with the endotracheal tube clamped at end-expiration. Lung function parameters were assessed by spirometry. A device for breath-by-breath monitoring metabolic gas exchange was used to measure global O(2) uptake. Relative impedance changes during ventilation and apnoea were related to the corresponding tidal volumes. Regional O(2) uptake was analysed as absolute values and as a ratio to regional ventilation in two regions of interest (ventral and dorsal). The global O(2) uptake measured by EIT was 208 +/- 79 ml min(-1) corresponding to the values obtained by metabolic gas exchange (259 +/- 73 ml min(-1); Spearman correlation coefficient: 0.81, p = 0.02). Regional O(2) uptake was significantly higher in the ventral lung region, while the regional O(2) uptake/ventilation ratio showed no significant difference between the regions. In conclusion, our pilot study indicates that EIT holds substantial potential to detect global and regional pulmonary O(2) uptake concordant with a linear lung volume decrease during apnoea. PMID- 21646715 TI - Front end with offset-free symmetrical current source optimized for time domain impedance spectroscopy. AB - Fast impedance measurements are often performed in time domain utilizing broad bandwidth excitation signals. Other than in frequency domain measurements harmonic distortion cannot be compensated which requires careful design of the analog front end. In order to minimize the influence of electrode polarization and noise, especially in low-frequency measurements, current injection shows several advantages compared to voltage application. Here, we show an active front end based on a voltage-controlled current source for a wide range of impedances. Using proper feedback, the majority of the parasitic capacitances are compensated. The bandwidth ranges from dc to 20 MHz for impedance magnitude below 5 kOmega. The output is a symmetric signal without dc-offset which is accomplished by combination of a current conveyor and a voltage inverter. An independent feedback loop compensates the offset arising from asymmetries within the circuitry. We focused especially on the stability of the current source for usage with small metal electrodes in aqueous solutions. At the monitor side two identical, high input impedance difference amplifiers convert the net current through the object and the voltage dropping across into a 50 Omega symmetric output. The entire circuitry is optimized for step response making it suitable for fast time domain measurements. PMID- 21646716 TI - Bioimpedance harmonic analysis as a tool to simultaneously assess circulation and nervous control. AB - Multicycle harmonic (Fourier) analysis of bioimpedance was employed to simultaneously assess circulation and neural activity in visceral (rat urinary bladder) and somatic (human finger) organs. The informative value of the first cardiac harmonic of the bladder impedance as an index of bladder circulation is demonstrated. The individual reactions of normal and obstructive bladders in response to infusion cystometry were recorded. The potency of multicycle harmonic analysis of bioimpedance to assess sympathetic and parasympathetic neural control in urinary bladder is discussed. In the human finger, bioimpedance harmonic analysis revealed three periodic components at the rate of the heart beat, respiration and Mayer wave (0.1 Hz), which were observed under normal conditions and during blood flow arrest in the hand. The revealed spectrum peaks were explained by the changes in systemic blood pressure and in regional vascular tone resulting from neural vasomotor control. During normal respiration and circulation, two side cardiac peaks were revealed in a bioimpedance amplitude spectrum, whose amplitude reflected the depth of amplitude respiratory modulation of the cardiac output. During normal breathing, the peaks corresponding to the second and third cardiac harmonics were split, reflecting frequency respiratory modulation of the heart rate. Multicycle harmonic analysis of bioimpedance is a novel potent tool to examine the interaction between the respiratory and cardiovascular system and to simultaneously assess regional circulation and neural influences in visceral and somatic organs. PMID- 21646717 TI - Loss of H3K4 methylation destabilizes gene expression patterns and physiological functions in adult murine cardiomyocytes. AB - Histone H3 lysine 4 (H3K4me) methyltransferases and their cofactors are essential for embryonic development and the establishment of gene expression patterns in a cell-specific and heritable manner. However, the importance of such epigenetic marks in maintaining gene expression in adults and in initiating human disease is unclear. Here, we addressed this question using a mouse model in which we could inducibly ablate PAX interacting (with transcription-activation domain) protein 1 (PTIP), a key component of the H3K4me complex, in cardiac cells. Reducing H3K4me3 marks in differentiated cardiomyocytes was sufficient to alter gene expression profiles. One gene regulated by H3K4me3 was Kv channel-interacting protein 2 (Kcnip2), which regulates a cardiac repolarization current that is downregulated in heart failure and functions in arrhythmogenesis. This regulation led to a decreased sodium current and action potential upstroke velocity and significantly prolonged action potential duration (APD). The prolonged APD augmented intracellular calcium and in vivo systolic heart function. Treatment with isoproterenol and caffeine in this mouse model resulted in the generation of premature ventricular beats, a harbinger of lethal ventricular arrhythmias. These results suggest that the maintenance of H3K4me3 marks is necessary for the stability of a transcriptional program in differentiated cells and point to an essential function for H3K4me3 epigenetic marks in cellular homeostasis. PMID- 21646718 TI - Mouse and human lung fibroblasts regulate dendritic cell trafficking, airway inflammation, and fibrosis through integrin alphavbeta8-mediated activation of TGF-beta. AB - The airway is a primary portal of entry for noxious environmental stimuli that can trigger airway remodeling, which contributes significantly to airway obstruction in chronic obstructive pulmonary disease (COPD) and chronic asthma. Important pathologic components of airway remodeling include fibrosis and abnormal innate and adaptive immune responses. The positioning of fibroblasts in interstitial spaces suggests that they could participate in both fibrosis and chemokine regulation of the trafficking of immune cells such as dendritic cells, which are crucial antigen-presenting cells. However, physiological evidence for this dual role for fibroblasts is lacking. Here, in two physiologically relevant models - conditional deletion in mouse fibroblasts of the TGF-beta-activating integrin alphavbeta8 and neutralization of alphavbeta8 in human COPD fibroblasts we have elucidated a mechanism whereby lung fibroblast chemokine secretion directs dendritic cell trafficking, in a manner that is critically dependent on alphavbeta8-mediated activation of TGF-beta by fibroblasts. Our data therefore indicate that fibroblasts have a crucial role in regulating both fibrotic and immune responses in the lung. PMID- 21646719 TI - Myeloid-derived suppressor cells are implicated in regulating permissiveness for tumor metastasis during mouse gestation. AB - Metastasis depends on the ability of tumor cells to establish a relationship with the newly seeded tissue that is conducive to their survival and proliferation. However, the factors that render tissues permissive for metastatic tumor growth have yet to be fully elucidated. Breast tumors arising during pregnancy display early metastatic proclivity, raising the possibility that pregnancy may constitute a physiological condition of permissiveness for tumor dissemination. Here we have shown that during murine gestation, metastasis is enhanced regardless of tumor type, and that decreased NK cell activity is responsible for the observed increase in experimental metastasis. Gene expression changes in pregnant mouse lung and liver were shown to be similar to those detected in premetastatic sites and indicative of myeloid cell infiltration. Indeed, myeloid derived suppressor cells (MDSCs) accumulated in pregnant mice and exerted an inhibitory effect on NK cell activity, providing a candidate mechanism for the enhanced metastatic tumor growth observed in gestant mice. Although the functions of MDSCs are not yet understood in the context of pregnancy, our observations suggest that they may represent a shared mechanism of immune suppression occurring during gestation and tumor growth. PMID- 21646720 TI - The LRF transcription factor regulates mature B cell development and the germinal center response in mice. AB - B cells play a central role in immune system function. Deregulation of normal B cell maturation can lead to the development of autoimmune syndromes as well as B cell malignancies. Elucidation of the molecular features of normal B cell development is important for the development of new target therapies for autoimmune diseases and B cell malignancies. Employing B cell-specific conditional knockout mice, we have demonstrated here that the transcription factor leukemia/lymphoma-related factor (LRF) forms an obligate dimer in B cells and regulates mature B cell lineage fate and humoral immune responses via distinctive mechanisms. Moreover, LRF inactivation in transformed B cells attenuated their growth rate. These studies identify what we believe to be a new key factor for mature B cell development and provide a rationale for targeting LRF dimers for the treatment of autoimmune diseases and B cell malignancies. PMID- 21646721 TI - Antagonism of miR-33 in mice promotes reverse cholesterol transport and regression of atherosclerosis. AB - Plasma HDL levels have a protective role in atherosclerosis, yet clinical therapies to raise HDL levels have remained elusive. Recent advances in the understanding of lipid metabolism have revealed that miR-33, an intronic microRNA located within the SREBF2 gene, suppresses expression of the cholesterol transporter ABC transporter A1 (ABCA1) and lowers HDL levels. Conversely, mechanisms that inhibit miR-33 increase ABCA1 and circulating HDL levels, suggesting that antagonism of miR-33 may be atheroprotective. As the regression of atherosclerosis is clinically desirable, we assessed the impact of miR-33 inhibition in mice deficient for the LDL receptor (Ldlr-/- mice), with established atherosclerotic plaques. Mice treated with anti-miR33 for 4 weeks showed an increase in circulating HDL levels and enhanced reverse cholesterol transport to the plasma, liver, and feces. Consistent with this, anti-miR33 treated mice showed reductions in plaque size and lipid content, increased markers of plaque stability, and decreased inflammatory gene expression. Notably, in addition to raising ABCA1 levels in the liver, anti-miR33 oligonucleotides directly targeted the plaque macrophages, in which they enhanced ABCA1 expression and cholesterol removal. These studies establish that raising HDL levels by anti miR33 oligonucleotide treatment promotes reverse cholesterol transport and atherosclerosis regression and suggest that it may be a promising strategy to treat atherosclerotic vascular disease. PMID- 21646722 TI - Virus-induced tumor inflammation facilitates effective DC cancer immunotherapy in a Treg-dependent manner in mice. AB - Vaccination using DCs pulsed with tumor lysates or specific tumor-associated peptides has so far yielded limited clinical success for cancer treatment, due mainly to the low immunogenicity of tumor-associated antigens. In this study, we have identified intratumoral virus-induced inflammation as a precondition for effective antitumor DC vaccination in mice. Administration of a tumor-targeted DC vaccine during ongoing virus-induced tumor inflammation, a regimen referred to as oncolysis-assisted DC vaccination (ODC), elicited potent antitumoral CD8+ T cell responses. This potent effect was not replicated by TLR activation outside the context of viral infection. ODC-elicited immune responses mediated marked tumor regression and successful eradication of preestablished lung colonies, an essential prerequisite for potentially treating metastatic cancers. Unexpectedly, depletion of Tregs during ODC did not enhance therapeutic efficacy; rather, it abrogated antitumor cytotoxicity. This phenomenon could be attributed to a compensatory induction of myeloid-derived suppressor cells in Treg-depleted and thus vigorously inflamed tumors, which prevented ODC-mediated immune responses. Consequently, Tregs are not only general suppressors of immune responses, but are essential for the therapeutic success of multimodal and temporally fine-adjusted vaccination strategies. Our results highlight tumor-targeting, replication competent viruses as attractive tools for eliciting effective antitumor responses upon DC vaccination. PMID- 21646724 TI - Is the port-access technique really safe as conventional perfusion extracorporeal circulation for all candidates of valve surgery? PMID- 21646723 TI - Absence of IFN-gamma accelerates thrombus resolution through enhanced MMP-9 and VEGF expression in mice. AB - Deep vein thrombosis (DVT) is a major cause of pulmonary thromboembolism, a leading cause of death in individuals with DVT. Several lines of evidence indicate proinflammatory cytokines such as TNF-alpha are involved in thrombus formation and resolution, but the roles of IFN-gamma remain unclear. To address this issue, we performed ligation of the inferior vena cava to induce DVT in WT and IFN-gamma-deficient (Ifng-/-) mice. In WT mice, intrathrombotic IFN-gamma levels were elevated progressively as the postligation interval was extended. Thrombus size was substantially smaller at 10 and 14 days in Ifng-/- mice than in WT mice. Intrathrombotic collagen content was remarkably reduced at more than 10 days after the ligation in Ifng-/- mice compared with WT mice. The expression and activity of MMP-9, but not MMP-2, was higher at the late phase in Ifng-/- mice than in WT mice. Moreover, intrathrombotic recanalization was increased in Ifng-/ mice, with enhanced Vegf gene expression, compared with that in WT mice. Activation of the IFN-gamma/Stat1 signal pathway suppressed PMA-induced Mmp9 and Vegf gene expression in peritoneal macrophages. Furthermore, administration of anti-IFN-gamma mAbs accelerated thrombus resolution in WT mice. Collectively, these findings indicate that IFN-gamma can have detrimental roles in thrombus resolution and may be a good molecular target for the acceleration of thrombus resolution in individuals with DVT. PMID- 21646725 TI - Left atrial distensibility and E/e' for estimating left ventricular filling pressure in patients with stable angina. -A comparative echocardiography and catheterization study-. AB - BACKGROUND: Although E/e' (the ratio of early diastolic mitral inflow velocity to early diastolic mitral annular velocity) is widely used to measure left ventricular filling pressure (LVFP), its accuracy is questionable in coronary artery disease patients. METHODS AND RESULTS: Echocardiograms and LVFP were obtained from 174 patients with stable angina (Canadian Cardiovascular Society angina grade I-II) who had received interventions for angiography-confirmed coronary stenosis. Compared with single-vessel groups, the multiple-vessel group exhibited lower mitral annular velocities, higher LVFP, and stronger correlations between E/regional e' and LVFP. Additionally, stronger correlations between E/regional e' and LVFP existed in patients with systolic dysfunction or lower variation of myocardial performance index (MPI) among anterior, inferior and lateral borders of mitral annulus. Average e' was not superior to any regional e' for assessing LVFP by the E/e' method. E/e' and left atrial (LA) ejection fraction (EF) correlated linearly with LVFP, but the correlation between LA distensibility and LVFP was logarithmical. Compared with E/e', LA distensibility and LAEF were superior for identifying high LVFP. CONCLUSIONS: E/e' is not completely satisfactory for assessing LVFP in patients with stable angina, especially those with single-vessel disease, preserved systolic function or high MPI variation. For identifying high LVFP, LA distensibility and LAEF are better than E/e'. PMID- 21646726 TI - Impact of the low- to high-density lipoprotein cholesterol ratio on composition of angiographically ambiguous left main coronary artery plaque. AB - BACKGROUND: A high low-density lipoprotein cholesterol (LDL-C) to a high-density lipoprotein cholesterol (HDL-C) ratio is associated with cardiac events, while the left main coronary artery (LMCA) is considered to be an important target of atherosclerotic plaque accumulation. This aim of the present study was to investigate the relationship between a LDL-C/HDL-C ratio and the characteristics of tissue components of LMCA plaque. METHODS AND RESULTS: One-hundred-twenty consecutive patients with stable angina pectoris who received chronic statin treatment underwent percutaneous coronary intervention for the left coronary artery. We prospectively performed integrated backscatter (IB) intravascular ultrasound (IVUS) to their LMCAs and evaluated the tissue characteristics. According to the median value of their LDL-C/HDL-C ratios (2.4), they were divided into 2 groups [high LDL-C/HDL-C ratio (>2.4) (n=60) or low LDL-C/HDL-C ratio (<= 2.4) (n=60)]. There was no significant difference in the data analyzed using conventional IVUS between the 2 groups. In the IB-IVUS analysis, patients with a high LDL-C/HDL-C ratio had a larger lipid volume and a smaller fibrous volume compared to patients with a low LDL-C/HDL-C ratio (52 +/- 10% vs. 48 +/- 10%, P=0.014 and 45 +/- 9% vs. 50 +/- 10%, P=0.010). CONCLUSIONS: A high LDL C/HDL-C ratio was associated with a high percentage of lipid volume and a low percentage of fibrous volume in LMCA lesions. Our findings might well suggest the increased risk of cardiovascular events in patients with a high LDL-C/HDL-C ratio. PMID- 21646727 TI - Possible targets of therapy for catecholaminergic polymorphic ventricular tachycardia. - Insight from a theoretical model -. AB - BACKGROUND: Catecholaminergic polymorphic ventricular tachycardia (CPVT) is a serious disease with a high mortality but its management is limited. The aim of this study was to investigate specific target sites for therapy in order to find potential management strategies for CPVT. METHODS AND RESULTS: The mutant Ryanodine receptor 2 (RyR2) with reduced stored-overloaded-induced Ca2+ release (SOICR) threshold was incorporated into the Luo-Rudy dynamic (LRd) cell model to elucidate the underlying pathologies of CPVT. The simulations reveal that beta adrenergic stimulation increased the Ca2+ load in cardiac myocyte, which facilitates spontaneous SR Ca2+ leakage, resulting in triggered arrhythmias. Varied blockade (from 0% to 90%) in specific ion channels, including the Na+/Ca2+ exchanger (I(NaCa)), fast Na+ channel (I(Na)), RyR2 receptor (I(rel)), Ca2+ ATPase (SERCA) (I(up)) or L-type Ca2+channel (I(Ca(L))),was performed to simulate the action of specific drugs on target sites. Blockade of the I(NaCa) (<= 10% blockade), in contrast to the I(up) (<= 30% blockade), I(Ca(L)) and I(Na) (<=40% blockade), and followed by I(rel) (<= 80% blockade), was most effective in suppressing the triggered arrhythmias in CPVT. Specifically, dual blockade of I(Ca(L))/I(up), I(Na)/I(rel) or I(Ca(L))/I(rel) had a synergistic effect in CPVT management. CONCLUSIONS: Blockade of I(NaCa) appears to be the most efficacious target for CPVT management. Dual blockade of I(Ca(L))/I(up), I(Na)/I(rel) or I(Ca(L))/I(rel) has a synergistic effect in CPVT treatment. PMID- 21646728 TI - Human cytochrome P450 1A1 is a novel target gene of liver X receptor alpha. AB - Cytochrome P450 (CYP) 1A1 is involved in the metabolic activation of polycyclic aromatic hydrocarbons (PAHs) and is induced by several compounds, including PAHs. The induction of CYP1A1 mediated by the aryl hydrocarbon receptor (AhR) has been well investigated; however, little has been reported on the mechanisms of CYP1A1 induction mediated by factors other than AhR. In this study, we investigated the involvement of liver X receptor alpha (LXRalpha) in the induction of CYP1A1. TO 901317, an LXRalpha ligand, induced CYP1A1 mRNA in a dose-dependent fashion. Luciferase reporter assays using HepG2 cells showed that TO-901317 was capable of activating the promoter of the CYP1A1 gene and that a direct repeat 4 (DR4) motif located in a region from -452 to -467 was required for the induction of CYP1A1 through LXRalpha. Specific binding of LXRalpha to this DR4 motif was confirmed by gel shift and chromatin immunoprecipitation assays. Co-treatment of HepG2 cells with TO-901317 and 2,3,7,8-tetrachlorodibenzo-p-dioxin, a typical AhR ligand, caused the synergistic induction of CYP1A1 mRNA. Thus, we propose that the expression of CYP1A1 is regulated by LXRalpha as well as by AhR, suggesting that exposure to both LXRalpha and AhR ligands can result in the alteration of individual susceptibility to environmental carcinogens metabolically activated by CYP1A1. PMID- 21646729 TI - The expression of ghrelin O-acyltransferase (GOAT) in human tissues. AB - Ghrelin is a circulating growth hormone-releasing and appetite-inducing brain-gut peptide. It needs to be acylated on its serine-3 with octanoate for its endocrine actions. The acyl-transferase that catalyses ghrelin octanoylation has recently been identified and named as GOAT (ghrelin O-acyltransferase); GOAT enzyme is coded by the MBOAT4 gene. This study aimed to investigate GOAT expression in the human. The distribution of GOAT mRNA expression was studied in various human tissues using classical and real-time reverse transcription and polymerase chain reaction. GOAT expression was found in all tissues studied (stomach, adrenal cortex, breast, right and left colon, duodenum, jejunum, ileum, fat, Fallopian tube, gallbladder, lymph node, lymphocyte cell line, kidney, liver, lung, muscle, myocardium, pituitary, oesophagus, pancreas, ovary, placenta, prostate, testis, spleen and thyroid). The widespread expression of GOAT corresponds to the widespread distribution of ghrelin expression. GOAT expression was high in stomach and gut, the major ghrelin-secreting tissues, and in the pituitary, in which ghrelin is known to show autocrine and paracrine effects. Identification of GOAT expression in various tissues support the concept that in addition to the important endocrine effect of acylated ghrelin, the paracrine effects of locally synthetised and acylated ghrelin may be important. PMID- 21646730 TI - Cushing's disease in a patient with steroid 21-hydroxylase deficiency. AB - Cushing's disease rarely appears as a consequence of hereditary disease. However, familial diseases with diminished glucocorticoid feedback are associated with secondary hypercorticotropinism and have been shown to give rise to pituitary adenomas. We here describe the rare case of a 30-year old female patient with congenital adrenal hyperplasia who also showed clinical signs and a typical history of hypercortisolism that was specified as Cushing's disease. After removal of a pituitary microadenoma, serum-cortisol levels fell below normal and the symptoms improved. However, after four years the menstrual cycle was irregular again and ACTH levels were in the upper range of normal. A corticotropin challenge showed a minor cortisol response but a marked increase in 17-hydroxyprogesterone serum concentrations. Genetic analysis revealed a homozygous mutation in exon 7 of the CYP21A2 gene (CTG>TTG, p.V281L). We conclude that a marked ACTH drive was able to override insufficient 21-hydroxylation and even to cause hypercortisolism. Although we describe a rare case, the impairment of the glucocorticoid feedback system in the context of congenital adrenal hyperplasia and other diseases may contribute to the development of secondary hypercorticotropinism as well as corticotropin producing adenomas. PMID- 21646731 TI - Relationship between vaspin gene expression and abdominal fat distribution of Korean women. AB - Visceral adipose tissue-derived serpin (vaspin) is a novel adipokine that is thought to have insulin-sensitizing effects. We investigated vaspin mRNA expression in abdominal adipose tissue and examined how gene expression related to abdominal fat distribution and metabolic parameters in Korean women. We measured anthropometric variables, metabolic parameters, serum vaspin concentration, and vaspin mRNA expression in abdominal adipose tissue obtained from women who underwent abdominal gynecological surgery and were aged 18-67 years (n = 85). Visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) area were measured in 40 subjects using computed tomography (CT). Vaspin expression was analyzed by real-time quantitative RT-PCR according to abdominal fat distribution. Vaspin mRNA expression was greater in adipocytes than in stroma/vascular cells. In the total subjects, vaspin expression was significantly higher in SAT than in VAT. Vaspin expression in SAT in subcutaneous fat type (VSR <= 0.3) was significantly higher than in visceral fat type (VSR > 0.3), although vaspin expression in VAT was similar between subcutaneous and visceral fat type. There was a significant negative correlation between vaspin expression in SAT and VAT area (r = -0.55, p = 0.001). Serum vaspin concentration was significantly correlated with fasting insulin (r = 0.30, p = 0.02), HOMA-IR (r = 0.29, p = 0.02), and the ratio of vaspin expression in VAT to vaspin expression in SAT (r = 0.41, p = 0.04). Vaspin expression in abdominal adipose tissue was adipocyte specific and vaspin expression in SAT decreased as VAT area increased. PMID- 21646732 TI - Effects of cardiologist experience on outcomes of patients with ST-elevated myocardial infarction treated with primary PCI in a local area in Japan. AB - Primary percutaneous coronary intervention (PCI) for ST-elevated myocardial infarction (STEMI) results in dramatically improved clinical outcomes when performed in a timely manner. Although guidelines for STEMI patients recommend PCI should be performed by experienced operators with acceptable PCI volume, cardiologists in a local area must perform primary PCI at their own hospitals. This study evaluated the effects of cardiologist experience on outcomes for STEMI patients in a local area in Japan.Between April 2007 and March 2010, 140 consecutive STEMI patients were admitted to our hospital and 121 of these patients received primary PCI. STEMI patients undergoing primary PCI were divided into two groups according to the operator's experience as a cardiologist. We retrospectively analyzed their clinical backgrounds, PCI findings, in-hospital outcomes, and drug administration at discharge.There were no significant differences in any clinical characteristics, angiographic findings, or PCI procedures between the two groups. Clinical outcomes of the two groups were similar, except for the length of hospital stay (21.1 +/- 5.8 versus 15.5 +/- 9.7; P = 0.0255). The frequency of administration of drugs such as beta-blockers (59.1% versus 34.0%; P = 0.0086), aldosterone blockade (10.4% versus 25.5%; P = 0.0334), and nicorandil (76.1% versus 25.5%; P = < 0.0001) was different between the two groups.The clinical outcomes of STEMI patients in this study were satisfactory and almost equivalent when compared according to the experience of the attending cardiologist. The present findings suggest the important clinical implication that younger cardiologists who have experienced PCI procedures to a certain extent can safely perform primary PCI and contribute to better prognoses of STEMI patients. PMID- 21646733 TI - The safety and feasibility of bilateral radial approach in chronic total occlusion percutaneous coronary intervention. AB - Few studies have reported results for transradial (TR) percutaneous coronary intervention (PCI) for chronic total occlusion (CTO) lesions. The purpose of this study was to evaluate the feasibility and safety of bilateral radial PCI for CTO lesions.Eighty-five consecutive patients with CTO lesions received PCI via a bilateral TR approach. A high radial artery puncture (10-15 cm above styloid process) accommodating a 7 Fr catheter (85 cm long) was used for a retrograde approach, and a 6 Fr catheter was used in the other radial artery for an antegrade approach. Retrograde wiring was conducted primarily or after failure of antegrade wiring. Mean duration of CTO was 42.8 +/- 54.9 months. Vessels with occlusions attempted were the left anterior descending artery (40.0%; 34/85), right coronary artery (58.8%; 50/85), and left circumflex artery (1/85). PCI re attempts were made in 41.2% of the cases. The overall success rate was 87.1%. Retrograde wiring was successful in 61/85 cases (71.8%), via septal collaterals followed by epicardial collaterals and saphenous vein graft. There were no major complications (30 day in-hospital death, Q wave myocardial infarction, or emergency bypass surgery), or serious access site complications.For experienced TR-PCI operators who are already doing complex TR coronary interventions, the bilateral radial approach for CTO lesions appears feasible and safe. PMID- 21646734 TI - Enhanced inflammation in epicardial fat in patients with coronary artery disease. AB - It has been hypothesized that epicardial fat, a local visceral fat depot with close proximity to coronary arteries, may serve as a source of inflammatory cytokines and cells in coronary atherosclerotic lesions. Here, we characterized infiltration of inflammatory cells and expression of adipocytokines in epicardial adipose tissue in patients with and without coronary artery disease (CAD). Pare samples were obtained from epicardial and subcutaneous adipose tissue during elective cardiac surgery (CAD, n = 8; non-CAD, n = 9). Inflammatory cell infiltration was investigated by immunohistochemical staining using antibodies against CD3, CD4, CD8 and CD68. Expression of adipocytokines was evaluated by real-time quantitative reverse transcription-polymerase chain reaction. Infiltration of macrophages and CD8-positive T cells in the epicardial adipose tissue in the CAD group was greater than that in the non-CAD group. In contrast, there was no significant difference between the two groups in the number of inflammatory cells in subcutaneous adipose tissue. No statistical difference could be found between the CAD group and the non-CAD group in the expression levels of adiponectin and inflammatory cytokines in epicardial adipose tissue. Our findings suggest that inflammatory cell infiltration is enhanced in epicardial adipose tissue, but not in subcutaneous fat, in patients with coronary artery disease. Chronic inflammation in epicardial fat may influence the pathogenesis of coronary atherosclerosis. PMID- 21646735 TI - Heart type fatty acid binding protein is more sensitive than troponin I and creatine kinase myocardial band at early stage in determining myocardial injury caused by percutaneous coronary intervention. AB - Measurement of circulating cardiac biomarkers has enabled early diagnosis and risk assessment of acute coronary syndrome. Heart type fatty acid binding protein (H-FABP) is a relatively novel marker for the diagnosis of myocardial injury. The purpose of the present study was to compare H-FABP with Troponin I (cTnI) and creatine kinase myocardial band (CK-MB) in determining myocardial injury in patients with early stage of percutaneous coronary intervention (PCI). Blood was withdrawn one hour before and 3 hours after PCI from 40 patients to measure H FABP, cTnI and CKMB. H-FABP was measured qualitatively. CK-MB and cTnI were measured by a sandwich enzyme-linked immunosorbent assay. Before PCI, H-FABP was found to be negative, while cTnI and CK-MB were found to be in normal ranges. Statistical analysis of measurements 3 hours after PCI revealed that H-FABP was significantly positive in 15 (37%) patients, while cTnI was elevated in 11 (27%) patients and CKMB was elevated in 8 (20%) patients. H-FABP is statistically more sensitive than cTnI and CK-MB at detecting myocardial injury after PCI.H-FABP can be used in early stages to detect myocardial injury caused by PCI. H-FABP is more sensitive than cTnI and CK-MB in determining myocardial injury due to PCI within 3 hours. H-FABP may help us stratify a patient's risk in early stages after PCI. PMID- 21646736 TI - Localisation of SCN10A gene product Na(v)1.8 and novel pain-related ion channels in human heart. AB - We have shown that the gene SCN10A encoding the sodium channel Na(v)1.8 is a susceptibility factor for heart block and serious ventricular arrhythmia. Since Na(v)1.8 is known to be present in nerve fibres that mediate pain, it may be related to both cardiac pain and dysrhythmia. The localisation of Na(v)1.8 and other key nociceptive ion channels, including Na(v)1.7, Na(v)1.9, capsaicin receptor TRPV1, and purinergic receptor P2X(3), have not been reported in human heart. The aim of this study was to determine the distribution of Na(v)1.8, related sodium and other sensory channels in human cardiac tissue, and correlate their density with sympathetic nerves, regenerating nerves (GAP-43), and vascularity. Human heart atrial appendage tissues (n = 13) were collected during surgery for valve disease. Tissues were investigated by immunohistology using specific antibodies to Na(v)1.8 and other markers. Na(v)1.8 immunoreactivity was detected in nerve fibres and fascicles in the myocardium, often closely associated with small capillaries. Na(v)1.8 nerve fibres per mm(2) correlated significantly with vascular markers. Na(v)1.8-immunoreactivity was present also in cardiomyocytes with a similar distribution pattern to that seen with connexins, the specialised gap junction proteins of myocardial intercalated discs. Na(v)1.5-immunoreactivity was detected in cardiomyocytes but not in nerve fibres. Na(v)1.7, Na(v)1.9, TRPV1, P2X(3)/P2X(2), and GAP43 positive nerve fibres were relatively sparse, whereas sympathetic innervation and connexin43 were abundant. We conclude that sodium channel Na(v)1.8 is present in sensory nerves and cardiomyocytes of human heart. Na(v)1.8 and other pain channels provide new targets for the understanding and treatment of cardiac pain and dysrhythmia. PMID- 21646737 TI - The prognostic value of atrial fibrillation on 30-day clinical outcome in patients with ST-segment elevation myocardial infarction undergoing primary percutaneous coronary intervention. AB - This study evaluated the association between atrial fibrillation (AF) and 30-day clinical outcome in patients with acute ST-segment elevation myocardial infarction (STEMI) undergoing primary percutaneous coronary intervention (PCI). Between January 2005 and October 2009, 783 consecutive patients with acute STEMI undergoing primary PCI were enrolled. Of these patients, 85 (10.9%) with AF during admission were categorized into group 1, while the remaining 698 (89.1%) with sinus rhythm during admission served as group 2. The results demonstrated that the incidence of advanced Killip score (defined as >= score 3) and advanced congestive heart failure (defined as >= NYHA class 3) were significantly higher, whereas the left ventricular ejection fraction (LVEF) was notably lower in group 1 than in group 2 (all P < 0.003). Additionally, the normal blood flow in the infarct-related artery was notably lower in group 1 than in group 2 (P = 0.003). Moreover, the incidences of new-onset stroke and 30-day mortality were remarkably higher in group 1 than in group 2 (all P < 0.003). Furthermore, Kaplan-Meier analysis demonstrated that the 30-day survival rate was markedly lower in AF patients than in those with sinus rhythm. However, multivariate stepwise Cox regression analysis demonstrated that the advanced Killip score and low LVEF were significantly and independently predictive of 30-day mortality (all P < 0.004). In conclusion, AF was significantly associated with 30-day mortality. PMID- 21646738 TI - A new criteria differentiating type 2 and 3 Brugada patterns from ordinary incomplete right bundle branch block. AB - The type 1 (coved) ECG pattern is diagnostic for Brugada syndrome; types 2 and 3 require antiarrhythmic drug challenge to confirm its presence. We evaluated a 12 lead ECG-based criterion to differentiate between ordinary incomplete right bundle branch block (iRBBB) and true type 2 and 3 patterns that evolve toward type 1 during drug challenge. The subjects were 22 patients (21 men, 1 woman; mean age, 46.8 +/- 13.2 years) referred for drug challenge (1 mg/kg pilsicainide, iv). In magnified ECG lead V1 and/or V2 with an iRBBB pattern, the baseline angle defined as the cross section of the upslope of the r' wave with the downslope of the r' wave was measured and compared between patients responding negatively versus positively to drug challenge, and was found to be significantly smaller in patients responding negatively (20.9 +/- 12.9 degrees , n = 6, versus 38.7 +/- 16.5 degrees , n = 13; P = 0.009). This ECG-based method successfully discriminates between the ordinary iRBBB pattern and drug-induced evolution toward a type 1 Brugada ECG. PMID- 21646739 TI - Elevated cardiac troponin T predicts adverse outcomes in hypertensive patients. AB - Ongoing myocardial damage detected as elevated serum cardiac troponin T (cTnT) indicates increased risk for future cardiac events in patients with chronic heart failure. Whether elevated cTnT is associated with adverse outcomes in patients with hypertension (HT) without left ventricular (LV) systolic dysfunction is unknown.We measured cTnT levels in 176 patients with essential HT without LV systolic dysfunction (LV ejection fraction <= 55%), renal failure, and prior cardiovascular or cerebrovascular diseases and 39 normal controls. Levels of cTnT were elevated (>= 0.02 ng/mL) in 15 (9%) of the 176 patients and in 0 (0%) of the 39 normal controls (P = 0.04). The rate of diabetes mellitus (DM), the cardiothoracic ratio, plasma B-type natriuretic peptide (BNP) value, and LV mass index were significantly higher in patients with than without elevated cTnT (DM, 8/15 versus 29/161, P = 0.004; cardiothoracic ratio, 54.5 +/- 4.5 versus 51.6 +/- 5.2%, P = 0.04; BNP, 103.3 +/- 142.3 versus 36.9 +/- 50.7 pg/mL, P = 0.04; LV mass index, 227 +/- 87 versus 152 +/- 57 g/m(2), P = 0.0001). Kaplan-Meier analysis demonstrated that significantly fewer (P < 0.000001) patients with, than without elevated cTnT remained free of events (hospitalization due to cardiovascular or cerebrovascular disease, n = 34). Stepwise Cox multivariate analysis revealed that elevated cTnT (hazard ratio, 6.58; P = 0.000001) and smoking (hazard ratio, 2.24; P = 0.04) were independent predictors of events.The present findings indicate that cTnT is a novel and useful predictor of future cardiovascular or cerebrovascular events in hypertensive patients. PMID- 21646740 TI - High mobility group box 1 levels in on and off-pump cardiac surgery patients. AB - High mobility group box 1 (HMGB1), which has properties similar to those of proinflammatory cytokines, is released from activated immune cells and necrotic cells. It is known that cardiopulmonary bypass (CPB) induces systemic inflammation and aortic cross-clamping induces myocardial ischemia. This study was conducted to clarify whether HMGB1 is released in CPB-supported cardiac surgery in comparison to off-pump coronary artery bypass grafting (OPCAB) where CPB is not used.Nineteen adult patients undergoing cardiac surgery involving CPB (CPB group) and 5 OPCAB patients (OPCAB group) were included in this study. Plasma concentrations of proinflammatory cytokines including HMGB1 were measured before, during, and after cardiac surgery. The plasma HMGB1 level was significantly increased at one hour after aortic declamping in the CPB group and at 30 minutes after revascularization in the OPCAB group. The peak HMGB1 level was slightly higher in the CPB group than that in the OPCAB group. These values decreased toward baseline value after surgery in both groups. TNF-alpha and IL 1beta were not detectable throughout the study period in either group. IL-6 and IL-10 increased after aortic declamping in the CPB group and after coronary revascularizations in the OPCAB group.Based on these results, we conclude that the major factor involved in the increase in HMGB1 level might be myocardial ischemia/reperfusion during cardiac surgery. Activation of immune cells, altered tissue perfusion, and pulmonary ischemia and reperfusion could be additional factors that increase the HMGB1 level in CPB-supported cardiac surgery. PMID- 21646741 TI - Virtual histology intravascular ultrasound compared with optical coherence tomography for identification of thin-cap fibroatheroma. AB - Virtual histology intravascular ultrasound (VH-IVUS) allows detailed assessment of plaque composition in the clinical setting. Optical coherence tomography (OCT) has been developed as a high-resolution imaging method, which might be a promising technique to identify thin-cap fibroatheroma (TCFA) in vivo. The purpose of the present study was to evaluate the diagnostic accuracy of VH-IVUS to identify TCFA as determined by OCT.We examined 96 target lesions in patients with stable angina pectoris by using VH-IVUS and OCT. VH-IVUS derived TCFA was defined as a focal necrotic core-rich lesion without evident overlying fibrous tissue. OCT derived TCFA was defined as a plaque with a fibrous cap of < 65 um. VH-IVUS correctly identified 16 TCFA and 67 non-TCFA but misclassified 2 TCFA and 11 non-TCFA as determined by OCT. The sensitivity, specificity, and positive and negative predictive values of VH-IVUS to identify TCFA as determined by OCT were 89%, 86%, 59%, and 97%, respectively.VH-IVUS showed an acceptable sensitivity and specificity to identify TCFA as determined by OCT. Although the positive predictive value was low reflecting a high number of false positives, the negative predictive value was notably high. Our results suggest a potential role for VH-IVUS to exclude high risk lesions for future coronary events. PMID- 21646742 TI - Serum C-reactive protein levels and death and cardiovascular events in mild to moderate chronic kidney disease. AB - Chronic kidney disease (CKD) is associated with an increased risk of cardiovascular (CV) disease. Elevated circulating levels of high sensitivity C reactive protein (hsCRP) have been suggested to be associated with high risk of CV disease. It is uncertain whether the CV risk in CKD can be stratified by hsCRP levels in the Japanese population. Baseline data including serum hsCRP and creatinine levels were determined in the general population. Estimated glomerular filtration rate (eGFR) was calculated using a modified MDRD equation, and CKD was defined as eGFR below 60 mL/minute/1.73m(2). We analyzed 1,074 male subjects with mild to moderate CKD (mean age, 70.4 years). CV events (stroke and myocardial infarction) and all-cause death were surveyed prospectively. The CKD subjects were followed for 5.1 years, and 72 CV events and 115 all-cause deaths were found (composite endpoint). After adjustment for established CV risk factors, hazard ratios (HRs) for the endpoint were significantly increased according to the hsCRP quintile (P < 0.001), and HR for the highest (versus the lowest) quintile was 2.77 (95% CI; 1.61-4.77). These results suggest that serum hsCRP measurement is a useful tool for the risk stratification of CV events and death in CKD male subjects selected from the general population. PMID- 21646743 TI - Effects of serum amyloid a and lysophosphatidylcholine on intracellular calcium concentration in human coronary artery smooth muscle cells. AB - Serum amyloid A (SAA), an acute-phase protein, and lysophosphatidylcholine (LPC), an oxidized LDL component, contribute to the physiological processes of atherosclerosis and cardiovascular disease. However, the effects of SAA/LPC on human coronary artery smooth muscle cells (hCASMCs) have not been fully investigated. Therefore, we examined the effects of SAA/LPC on Ca(2+)/Mg(2+) mobilization and its underlying mechanisms in hCASMCs. Intracellular Ca(2+)/Mg(2+) concentration ([Ca(2+)](i) / [Mg(2+)](i)) was measured with fura-2 AM/mag-fura-2 AM. Conventional RT-PCR analysis was also performed. Both SAA and LPC increased [Ca(2+)](i) by Ca(2+) entry. The SAA-induced Ca(2+) entry was inhibited by Gd(3+), SKF96365, and 2-aminoethoxydiphenyl borate (2-APB), a nonselective transient receptor potential (TRP) channel blocker, but not nifedipine. The LPC-induced Ca(2+) entry was blocked by Gd(3+), but not nifedipine, SKF96365 and 2-APB. U-73122 and PTX prevented the activation of SAA-, but not LPC-induced Ca(2+) influx. LPC, but not SAA, increased [Mg(2+)](i) as well as [Ca(2+)](i). The RT-PCR analysis revealed the expression of TRPC1/4, TRPV1/2/4, and TRPM7/8 mRNA. These results suggest that SAA/LPC activate Ca(2+) influx in hCASMCs; SAA activates it via PTX-sensitive G-protein, PLC and TRPC pathways, while LPC activates it independently of these pathways, where TRPM7 may be partly involved. Thus, TRP protein appears to be a target molecule of Ca(2+) signaling in hCASMCs elicited by SAA/LPC, which may play roles in coronary muscle dysfunction under pathophysiological and inflammatory conditions such as atherosclerosis. PMID- 21646744 TI - Heart failure mimicking prior myocardial infarction in a patient with idiopathic hypereosinophilic syndrome. AB - Idiopathic hypereosinophilic syndrome (IHES) is characterized by sustained, nonreactive hypereosinophilia with eosinophilia-associated organ damage. Cardiac involvement occurs in about 60% of patients with HES and it is the major cause of mortality in these patients. Cardiac dysfunction is reversible only after early corticosteroid (CS) initiation.Herein we report a 33-year old male who was referred to our Cardiology Department with electrocardiographic and echocardiographic abnormalities suggesting myocardial infarction. At presentation he complained of dyspnea, cough and persistent fever. His white blood cell (WBC) count was elevated, with eosinophil predominance in the differential. After cardiological and haematological work-up, the final diagnosis of HES-associated cardiac involvement was established. Early treatment with CS led to eosinophil count normalization with only moderate cardiac function improvement. Currently, the patient is in good condition overall and is in NYHA class II while still on prednisone. PMID- 21646745 TI - Prevalence of tinnitus in community-dwelling Japanese adults. AB - BACKGROUND: Several studies have reported the prevalence of tinnitus among general populations; however, most of these studies were conducted in Europe or the United States. We estimated the prevalence of tinnitus among the general adult population in Japan. METHODS: The subjects were participants in the Takayama Study, a population-based cohort study. In 2002, a total of 14 423 adults (6450 men and 7973 women) aged 45 to 79 years responded to a self administered questionnaire that inquired about history of tinnitus, which was defined as episodes lasting longer than 5 minutes, excluding those occurring immediately after noise exposure. Respondents were also asked about the loudness and severity of tinnitus. RESULTS: Overall, 11.9% of the subjects reported having tinnitus; the percentage was somewhat higher among men (13.2%) than women (10.8%). The prevalence of tinnitus increased with age in both sexes. Approximately 0.4% of the overall population reported that tinnitus had a severe effect on their ability to lead a normal life. Medical history of hypertension or ischemic heart diseases, use of steroid or antihypertensive medication, and employment as a factory worker or machine operator were associated with tinnitus status in both men and women. CONCLUSIONS: Tinnitus is relatively common in Japan. Although the use of various definitions of tinnitus in different studies makes it difficult to compare prevalence among populations, the present prevalence estimate was similar to those in studies in Europe and the United States. PMID- 21646747 TI - Hyperbaric air therapy in dogs for clinical veterinary medicine: a basic study. AB - Hyperbaric air therapy (HBA) is a treatment in which an animal is exposed to air pressurized to about 1.3 atmosphere absolute (ATA). Although HBA has already been administered to humans in medical applications, it has not been reported in clinical veterinary medicine. Therefore, we aimed to determine a safe protocol for dogs. To elucidate oxygen dynamics during HBA, we measured partial pressure of arterial oxygen, oxygen saturation of tissue, and partial pressure of transcutaneous oxygen in dogs. HBA could be performed safely with a protocol of pressurizing speed up to 0.1 ATA/min, maximum chamber pressure of up to 1.3 ATA, and pressure duration of around 45 min per treatment. Under these conditions, tissue was adequately oxygenated during and after treatment. PMID- 21646746 TI - Transmission of pandemic influenza A (H1N1) virus in a train in China. AB - BACKGROUND: Pandemic influenza A (H1N1) virus emerged in North America in April 2009 and spread globally. We describe the epidemiology and public health response to the first known outbreak of 2009 H1N1 in a train, which occurred in June 2009 in China. METHODS: After 2 provinces provided initial reports of 2009 H1N1 infection in 2 persons who had travelled on the same train, we conducted a retrospective epidemiologic investigation to collect information from the passengers, crew members, contacts, and health care providers. We explored the source of infection and possible routes of transmission in the train. All cases were confirmed by real-time reverse transcription polymerase chain reaction testing. RESULTS: Train #1223 traveled 40 hours, made 28 stops in 4 Chinese provinces, and boarded 2,555 passengers, who logged a total of 59,144 person hours of travel time. Nineteen confirmed 2009 H1N1 cases were identified. Of these, 13 were infected and developed symptoms on the train and 6 occurred among contacts who developed illness during medical monitoring. In addition, 3 asymptomatic cases were identified based on RT-PCR testing of respiratory swabs from contacts. The attack rate among contacts of confirmed cases in the same car was higher than that among contacts in other cars (3.15% vs. 0%, P < 0.001). Attack rates increased with exposure time. CONCLUSIONS: Close contact and long exposure may have contributed to the transmission of 2009 H1N1 virus in the train. Trains may have played an important role in the 2009 influenza pandemic. PMID- 21646748 TI - Cerebral vascular hamartoma with thrombosis in a dog. AB - A cerebral vascular hamartoma was identified in the frontal lobe, striatum and thalamus of the right side of the brain of a male, 7-year-old Shih Tzu. Histologically, the lesion consisted of thin-walled vessels, which showed various sizes and occasionally contained fibrin thrombi. These vascular walls were composed of a single layer of fibromuscular tissue lined by flat endothelium with various amount of collagen, but devoid of large coat of smooth muscles and elastic tissue. Immunohistochemically, the lining endothelial cells were positive for von Willebrand Factor antibody. Neuropil between the vessels was stained with Kluver-Barrera stain, and positive for synaptophysin and GFAP antibodies. Based on these findings, the lesion was diagnosed as vascular hamartoma, which might resemble venous malformation in humans. PMID- 21646749 TI - Possible origin of CSF antibodies induced by intrathecal immunization and prophylactic effects against intracerebral rabies virus infection. AB - Intrathecal (IT) immunization involves injecting antigens directly into the intraventricular or subarachnoid spaces, or brain, to induce antigen-specific antibodies (Ab) in the cerebrospinal fluid (CSF). In the present study, rabbits were immunized IT with inactivated rabies virus to investigate the origins of CSF Ab. The time course of Ab induction and tumor necrosis factor-alpha expression suggested the possibility that the CSF Ab originated in the serum. In addition, Ab-producing cells infiltrated around the blood vessels of the brain, suggesting local production of Ab within the central nervous system (CNS). Furthermore, subcutaneous (SC) immunization prior to IT immunization induced a rapid and magnified Ab response in the CSF compared with IT immunization alone. These results were confirmed by the fact that mice immunized SC prior to IT were more resistant to intracerebral challenge with rabies virus than mice immunized via the IT route alone. Taken together, these results suggest that combined SC and IT immunization is a more effective vaccination protocol for prophylaxis and treatment of rabies. PMID- 21646750 TI - Amino acid substitution at position 95 in rabies virus matrix protein affects viral pathogenicity. AB - We previously reported that rabies virus strain CE(NiM), but not the parental Ni CE strain, killed mice after intracerebral inoculation. CE(NiM) and Ni-CE are genetically identical except for two amino acids at positions 29 and 95 in the M protein. In this study, to identify which residue determines the pathogenicity, we examined pathogenicities of two Ni-CE mutants, CE(NiM29) and CE(NiM95), which were established by replacement of an amino acid residue at position 29 or 95 in the Ni-CE M protein with the corresponding residue of CE(NiM), respectively. We found that CE(NiM95), but not CE(NiM29), killed mice, indicating that the amino acid at position 95 in the M protein is the pathogenic determinant. PMID- 21646751 TI - Screening of therapeutic targets for canine mast cell tumors from a variety of kinase molecules. AB - Options of systemic treatment for canine MCT have been still limited and most canine cases with MCTs eventually undergo relapses even after achievement of a remission. Thus additional therapies are required to establish for the tumor. To identify the novel candidate therapeutic targets for canine MCT, the mRNA expression and phosphorylation statuses of several receptor or non-receptor kinases as well as the inhibitory effect of 95 specific inhibitors on the growth were assessed in three canine MCT cell lines (HRMC, VIMC1 and CMMC1). Among the 14 targets, the mRNAs of 11, 7 and 7 kinases were amplified in HRMC, VIMC1 and CMMC1, respectively. The mRNAs of VEGFR3, PDGFRalpha, SRC, YES, LCK and FYN were detected in all cell lines. The phosphorylation of 12, 8 and 7 kinases was observed by using specific antibody arrays in HRMC, VIMC1 and CMMC1, respectively. DTK, EPHB6, AMPKalpha1, CREB, STAT5a and STAT5b were phosphorylated in all cell lines. The 10, 9 and 17 inhibitors exhibited the biological activity against the growth of HRMC, VIMC1 and CMMC1, respectively. Only three inhibitors such as SB218078 (for Chk1), PDGF RTK inhibitor IV (for PDGFR) and radicicol (for Hsp90) suppressed the growth of all three cell lines. The present study indicated that several kinases, such as Chk1, PDGFR and Hsp90, could be used as therapeutic targets in the treatment for canine MCT. Further studies and clinical trials are warranted to apply the inhibitors for the treatment of the tumor. PMID- 21646752 TI - Molecular phylogenetic analysis of Theileria species detected from Japanese serows (Capricornis crispus) in Iwate Prefecture, Japan. AB - In the present study, we tried to detect DNA for ribosomal RNA genes of piroplasma parasites from the liver or blood of 43 Japanese serows (Capricornis crispus) in Iwate Prefecture of Japan by polymerase chain reaction. Approximately 500-bp amplicons were obtained in 35 (81.4%) of the 43 samples by amplification for V4 hyper-variable regions of the 18S rRNA gene, and the amplicons were considered to be DNA of Theileria species. The complete nucleotide sequences (1,700 or 1709 bp) of the 18S rRNA gene were determined in 20 samples and were divided into 5 genotypes that were phylogenetically separated into two different lineages showing a polyphyletic relation. The Theileria DNAs of the two different lineages were considered to be those of distinct species. PMID- 21646753 TI - Polled Intersex Syndrome with urethral atresia in a goat. AB - A 5-day-old hornless goat was referred with dysuria since birth. The scrotum was absent, and a small penis-like structure was seen below the perineal raphe. On the laparotomy, the testicles were found near the inguinal ring- and attached to a uterus-like structure. On histological analysis, the uterus-like structure was blind-end. Germ cells were absent in the testis. The karyotype of this goat was 60, XX and the SRY gene was absent. The goat was homozygous for a DNA deletion responsible for the Polled Intersex Syndrome (PIS). To the authors' knowledge, this is the first report as the clinical case of the PIS-/- goat with urethral atresia. PMID- 21646754 TI - Epidemiological and morphological studies of double-chambered right ventricle in dogs. AB - The double-chambered right ventricle (DCRV) is a rare congenital cardiac disease in dogs, and its detailed epidemiological and morphological features are not clearly understood. By investigating the profile, clinical signs, and characteristics of examination findings of eleven dogs with DCRV by means of a retrospective study, we attempted to clarify the epidemiology and morphology of the condition. The study group consisted of nine males and two females. Breeds included Pug (n=3), Miniature Dachshund (n=1), French Bull-dog (n=1), Shiba (n=1), and Retrievers (n=5). The attachment site of the anomalous muscular bundle was continuous with the cardiac apex in nine dogs, and it was attached to the right ventricle free wall in the other two dogs. In dogs with DCRV, at least one of the following conditions was present concurrently: congenital or acquired tricuspid valve regurgitation (TR), ventricular septal defect, and atrial septal defect. Also, the pressure difference between the two chambers increased over time, and progressive right-sided heart failure was observed. In summary, DCRV occurs in small breeds of dog as well as in large breeds of dog and it may be more prevalent in males. The existence of two types of DCRV in dogs was established. Dog with DCRVs will have a high incidence of concurrent cardiac abnormalities. Concurrent TR may be either congenital or acquired. DCRV is a congenital disorder, but the clinical condition progresses as the dog develops. PMID- 21646755 TI - Diffuse alveolar damage in a young cat. AB - A 10-month-old cat was diagnosed with congenital subvalvular aortic stenosis. To resolve its hypoxia, oxygen therapy was administered a couple of times a week during two months. The oxygen partial pressure in the chamber was maintained between 30 and 35%, and the time for one procedure was 12-24 hr. The animal died due to severe respiratory failure. At necropsy, the lungs were voluminous and had a rubbery texture. Histologically, large type II pneumocytes with occasional atypia had diffusely proliferated within the lungs. Interstitial fibrosis was not observed, although some alveolar septa were thickened along with fibrinous exudates and neutrophilic infiltration. The histology of these lesions was consistent with diffuse alveolar damage (DAD), which might have been partially due to oxygen toxicity. PMID- 21646756 TI - [Clinical applications of optical equipment-based image enhanced endoscopy within the upper gastrointestinal tract]. PMID- 21646757 TI - [Current status and future prospect of three-dimensional computed tomographic (CT) imaging for gastrointestinal tract diagnoses]. PMID- 21646758 TI - [Noninvasive assessment of liver fibrosis by measurement of stiffness in patients with chronic liver diseases]. PMID- 21646759 TI - [Advances in imaging modalities of hepatocellular carcinoma]. PMID- 21646760 TI - [A study of the usefulness of pancreatic juice cytology obtained via an endoscopic nasal pancreatic drainage (ENPD) tube]. AB - We studied the usefulness of pancreatic juice cytology obtained via an indwelling endoscopic nasal pancreatic drainage (ENPD) tube. In general, cytology was performed three times. The sensitivity was 0.35 on the first time and 0.59 after three times (p<0.01). The sensitivity in relation to tumor size of pancreatic cancer was 0.77 for Tis (3 cases) and TS1 (10 cases), 0.76 for TS2 (29 cases), 0.56 for TS3 (9 cases) and 0 for TS4 (4 cases). A significant difference of p=0.01 was recognized among the 4 groups, and the sensitivity for small tumors was higher than that for large tumors. The pancreatic juice can be obtained repeatedly via the ENPD tube and that contributes to improving the diagnostic accuracy. It is useful as a definitive diagnosis method in early stage pancreatic cancer because it is easier to detect positive results in smaller tumor, furthermore, it is possible to diagnose carcinoma in situ. PMID- 21646761 TI - [Two cases of early gastric cancer arising in gastric hyperplastic polyps]. AB - We report 2 cases of early gastric cancer arising in hyperplastic polyps. The first case was 6mm in diameter, tub1, in an intestinal phenotype. The second case was 10mm in diameter, pap, in a gastric phenotype. Before resection, their biopsies had been diagnosed as atypical hyperplastic polyps. These cases suggest that small gastric hyperplastic polyps possess a certain malignant potential. If a hyperplastic polyp shows atypia in the biopsy, a complete resection should be considered. PMID- 21646762 TI - [A family with attenuated familial adenomatous polyposis identified because of fundic gland polyposis]. AB - We report a 41-year-old woman who had fundic gland polyposis without a colorectal polyp. Because her father had colon cancer, multiple colorectal polyps and the gastric polyposis, we suspected AFAP and researched the APC gene. There was 1 base pair deletion of guanine at codon 99-100 in exon 3 of the APC gene, and its frame shift mutation made stop codon at codon 124. It was proved that the gene carrier of AFAP can be discovered based on the family-history and the presence of fundic gland polyposis, even when no colorectal polyps exist. PMID- 21646763 TI - [Significant regression of a cavernous hepatic hemangioma to a sclerosed hemangioma over 12 years: a case study]. AB - A sixties woman was found to have diagnosed by abdominal ultrasonography with a tumor in the left lobe of the liver and was referred to our institution in 1998. Abdominal magnetic resonance imaging (MRI) showed a typical, 70*45 mm cavernous hemangioma, which was followed up by annual MRI. In 2006, 8 years after the initial diagnosis, the MRI showed that the tumor had reduced to 30*15 mm. Although atypical of hemangioma, review of the annual observations indicated a diagnosis of regressive hemangioma, which also accorded with clinical observations. In 2009, a liver biopsy was performed by laparotomy during gastrectomy for gastric cancer. Pathological examination of the biopsy revealed sclerosed hemangioma tissue, confirming the diagnosis of regression of a cavernous hemangioma to a sclerosed hemangioma over 12 years. PMID- 21646764 TI - [A case of serous cystadenoma of the pancreas associated with obstructive jaundice during long-term follow up]. AB - A 68-year-old man had been followed up since March, 1997 because of a cystic tumor of the pancreas head. The patient developed obstructive jaundice and was admitted to our hospital in June, 2007. The tumor size on CT scan had increased from 3.6 cm to 5.9 cm during the 10-year period. After endoscopic biliary drainage, pancreatoduodenectomy was performed. Pathological diagnosis of the resected specimen was serous cystadenoma. Serous cystadenoma of the pancreas is known as a benign tumor with indolent progression and is likely to be symptomatic if the tumor size exceeds 4 cm. However, biliary obstruction is a rare complication of serous cystadenoma. We report this rare case here with references to the literature. PMID- 21646765 TI - [A case of IgG4-related sclerosing disorders involving the mesentery and the pancreas]. AB - A 78-year-old man presented with upper abdominal pain. He underwent an abdominal computed tomography scan which revealed irregularly shaped mass lesions in the mesentery and in the pancreatic head and body. The mass lesions were suspected to be pancreatic cancer with peritoneal dissemination and a surgical biopsy was performed. Histologic studies revealed lymphoplasmacytic sclerosing pancreatitis with significant infiltration of IgG4-positive plasma cells. His serum IgG4 level was 223 mg/dl. Findings from histologic and serum studies were compatible with IgG4-related sclerosing disorders. The mass lesions responded to steroid therapy and disappeared. The difficulty in making a definitive diagnosis is discussed. PMID- 21646766 TI - [Ruptured aneurysm of a posterior inferior pancreaticoduodenal artery associated with duodenal stenosis after transcatheter arterial embolization]. AB - We describe a 72-year-old man admitted to hospital as an emergency case of epigastric abdominal pain. CT scan visualized massive hemorrhage around the pancreatic head. Computed tomographic angiography showed stenosis at the origin of the celiac artery and a 10 mm aneurysm of the posterior inferior pancreaticoduodenal artery (PIPDA). An emergency angiogram revealed a long aneurysm in the PIPDA. The aneurysm had irregular width and was 75 mm in length. A gastroduodenal artery and the PIPDA were supplied from the superior mesenteric artery. A transcatheter arterial embolization (TAE) was performed. We reviewed 45 cases of pancreaticoduodenal aneurysms after 2000 and cases of the pancreaticoduodenal false aneurysms after 1972. As a result, we inferred that this case without pancreatitis or pancreas surgery was a true aneurysm made by the bloodstream changes caused by the celiac artery stenosis. PMID- 21646767 TI - [Metastasis of uterine leiomyosarcoma to the pancreas--usefulness and limitations of EUS-FNA]. AB - A woman in her 60's presented with a tumor of the pancreatic body. Pan hysterectomy had been performed under a diagnosis of uterine leiomyoma 11 years previously. A sample obtained by endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) revealed the histopathological proliferation of spindle shaped bundles of atypical cells, and immunohistochemical staining demonstrated that these cells were positive for KIT. Therefore, distal pancreatectomy was performed under a diagnosis of pancreatic gastrointestinal stromal tumor (GIST). Immunohistochemical staining of surgical specimens demonstrated that the tumor cells were positive for desmin and negative for KIT and CD34. The low-grade leiomyosarcoma in pathological specimens of the uterine myoma obtained 11 years previously histologically resembled the pathological findings of the pancreatic specimens except for atypical nuclei and mitotic cells. Therefore, the final diagnosis was extremely rare metastatic leiomyosarcoma of the pancreas. Herein, we report metastasis of uterine leiomyosarcoma to the pancreas and discuss the usefulness and limitations of EUS-FNA. PMID- 21646768 TI - [WT1 peptide-based immunotherapy for the treatment of malignancies: focusing on hematological neoplasms]. PMID- 21646769 TI - [Cellular immunotherapy with genetically engineered T-cell therapy for malignancies]. PMID- 21646770 TI - [Molecular pathogenesis by human T-cell leukemia virus type 1]. PMID- 21646771 TI - [Aberrant expression of microRNA and its role in malignant lymphoma]. PMID- 21646772 TI - [Remission induced by dose-reduction of immunosuppressants alone in a patient with post-transplant lymphoproliferative disorder of central nervous system origin]. AB - A 43-year-old male renal transplant recipient, who received a living related renal transplant 7 years ago and had been maintained with tacrolimus, mycophenolate mofetil (MMF), and prednisolone, was admitted to our hospital complaining of headache and nausea. MRI showed a large mass in the right hemisphere with ring-enhancement indicating brain abscess, tumor or lymphoma. Open biopsy was performed and pathological examination demonstrated diffuse proliferation of polymorphic cells, positive for CD20, bcl-2, EBER, and LMP-1. Based on these findings, primary central nervous system post-transplant lymphoproliferative disorder (PCNS-PTLD) was diagnosed. MMF was discontinued and tacrolimus was tapered. After 2 weeks, MRI showed regression of the tumor size and after 9 months, the tumor had disappeared. Though many reports have shown the severity of PCNS-PTLD, and recommend aggressive treatments such as chemotherapy and/or radiotherapy, our case shows that reduction of immunosuppressant alone with close observation could be a choice of treatment. PMID- 21646773 TI - [Chronic myelogenous leukemia complicated by drug-induced agranulocytosis]. AB - We describe a patient with chronic myelogenous leukemia (CML) who developed drug induced agranulocytosis. A 75-year-old female was diagnosed with CML in December 2001. She had been receiving imatinib therapy for more than five years. In August 2007, she was hospitalized due to a severe neutropenia 10 days after colonoscopy. She was diagnosed as having agranulocytosis induced by colonoscopy premedication including scopolamine butylbromide and flumazenil. Severe neutropenia was resolved by G-CSF treatment without CML progression. Agranulocytosis in patients with CML is rare, but potentially lethal. Here, we report the clinical course in this patient. PMID- 21646774 TI - [Hemorrhagic colitis caused by dasatinib following cytomegalovirus enterocolitis in a patient with chronic myelogenous leukemia in the second chronic phase]. AB - A 26-year-old female progressed to blastic crisis (BC) after three months administration of imatinib for chronic myelogenous leukemia (CML) chronic phase (CP) and was treated with a dasatinib containing chemotherapy regimen. After remission to second CP, she was hospitalized because of fever and hemorrhagic diarrhea during dasatinib maintenance therapy. She was diagnosed as having cytomegalovirus (CMV) colitis because CMV antigen in blood leukocytes was positive and CMV-positive cells were also detected on staining of an ileocecal mucosal biopsy specimen with an anti-CMV antibody. Although blood leukocyte CMV antigen and CMV staining in colonic mucosa became negative after ganciclovir treatment, hemorrhagic diarrhea did not improve. However, after discontinuance of dasatinib, hemorrhagic colitis drastically improved and did not recur after administration of nilotinib. It is possible that hemorrhagic diarrhea occurred due to dasatinib-related hemorrhagic colitis. Previous case reports have indicated that CD8-positive T-lymphocytes infiltrate the colonic mucosa in dasatinib-related hemorrhagic colitis, and the same pathological findings were seen in our case. Dasatinib may cause hemorrhagic colitis via immunological mechanisms in CML. Dasatinib-related gastrointestinal bleeding is less frequent in Japan compared to that in western countries, and Japanese cases diagnosed as having hemorrhagic colitis are extremely rare. PMID- 21646775 TI - [Acute lymphoblastic leukemia complicated with varicella zoster virus meningoencephalitis and visceral dissemination after related bone marrow transplantation]. AB - Meningitis or encephalitis by varicella-zoster virus (VZV) after hematopoietic stem cell transplantation (HSCT) is rarely reported. We encountered a case of meningoencephalitis with VZV re-activation 18 months after related bone marrow transplantation for recurrent acute lymphoblastic leukemia. The patient had been administered steroid and cyclosporine for chronic graft-versus-host disease. A high DNA copy number of VZV, 4.9*10(7) copies was detected in the cerebrospinal fluid. VZV also caused severe pneumonia and acute renal failure soon after the onset of meningoencephalitis. The patient was successfully treated with acyclovir, although he was left with persistent neurological sequelae. Both prompt diagnosis and early treatment of VZV reactivation are important to avoid a fatal outcome. PMID- 21646776 TI - [Successful treatment with immunosuppressive therapy for aplastic anemia developed after resection of thymoma]. AB - We report here a very severe case of thymoma-related aplastic anemia that developed after thymectomy. The patient was a 50-year-old man diagnosed with myasthenia gravis. Chest CT showed thymoma measuring 7.5 cm in diameter, and extended thymectomy was performed. Irradiation to the anterior mediastinum was added postoperatively. Thirteen months after surgery, hemogram showed severe neutropenia: leukocyte count 0.32*10(9)/l with 11% neutrophils; Hb 10.7 g/dl; and platelet count 100*10(9)/l. Although cyclosporine (CSP, 5 mg/kg/day) was administered, dose reduction was necessary because of renal damage. Cytopenia deteriorated to a leukocyte count of 0.71*10(9)/l with 21% neutrophils; Hb 5.9 g/dl; and platelet count 24*10(9)/l. However, addition of antithymocyte globulin (ATG, 15 mg/kg) led to hematopoietic recovery of all three lineages within one month. He is clinically stable with no transfusion requirement after 22 months with CSP maintenance therapy. Although thymoma-related aplastic anemia has been reported to have an extremely poor prognosis, high efficacy of CSP has been reported recently. In our case, ATG in combination with CSP was efficient. Adequate immunosuppressive therapy seems to be important for the clinical management of these patients. PMID- 21646777 TI - Isolation and characterization of murine mandibular condylar cartilage cell populations. AB - OBJECTIVES: The mandibular condylar cartilage is a heterogeneous tissue containing cells at various stages of chondrocyte maturation organized into 4 zones: superficial, polymorphic, flattened, and hypertrophic. The goal of this study was to use transgenic mice containing chondrocyte maturation markers fused to fluorescent protein transgenes to isolate and characterize homogenous cell populations of the mandibular condylar cartilage. METHODS: Fluorescent reporter expression in the mandibular condylar cartilage of transgenic mice containing the 3.6-kb fragment of the rat collagen type 1 promoter fused to a topaz-fluorescent protein (Col3.6-tpz), collagen type 2 promoter fused to a cyan-fluorescent protein (Col2-cyan), and/or collagen type 10 promoter fused to cherry-fluorescent protein (Col10-cherry) was examined. Mandibular condylar cartilage cells were analyzed by fluorescence-activated cell sorting (FACS) and either used for gene expression analysis or plated in cell cultures and exposed to adipogenic, osteogenic, or chondrogenic conditions. To determine cell fate, transgenic mice containing the Col3.6-cre recombinase were bred with cre reporter mice. RESULTS: Localization and analysis of gene expression revealed that Col3.6-tpz-positive cells corresponded to the polymorphic/flattened zones and Col2-cyan-positive cells corresponded to the flattened/hypertrophic zones of the mandibular condylar cartilage. Mandibular condylar cartilage FACS-sorted Col3.6-tpz-positive cells have the potential to differentiate into bone, cartilage, and fat. Cell fate mapping revealed that Col3.6 cells are precursors of some of the hypertrophic chondrocytes in the mandibular condylar cartilage. CONCLUSION: Col3.6-tpz cells represent an earlier stage of the mandibular condylar cartilage maturation pathway. PMID- 21646778 TI - Hypoplastic myelodysplastic syndrome associated with der(1;7)(q10;p10) presenting as bone marrow failure. PMID- 21646779 TI - Elevated levels of triglyceride and triglyceride-rich lipoprotein triglyceride induced by a high-carbohydrate diet is associated with polymorphisms of APOA5 1131T>C and APOC3-482C>T in Chinese healthy young adults. AB - BACKGROUND/AIMS: Changes in lipid profiles have been shown to be associated with diet and apolipoprotein (APO) polymorphisms. Therefore, 2 polymorphisms, i.e. APOA5-1131T>C and APOC3-482C>T, and serum lipids were examined in a Chinese healthy young population with high-carbohydrate/low-fat (HC/LF) diet intervention. METHODS: After a wash-out diet for 7 days, 56 young adults (22.89 +/- 1.80 years) received the HC/LF diet for 6 days. Body mass index (BMI) and fasting serum lipid profiles at baseline, after the wash-out diet, and after the HC/LF diet were measured. RESULTS: APOA5-1131C carriers had higher triglyceride (TG) and TG-rich lipoprotein TG (TRL-TG) levels at baseline and after the HC/LF diet, though this mainly corresponded to the female cohort. APOC3-482T carriers had higher TRL-TG levels following the wash-out and HC/LF diets, but these were not directly attributable to a single gender. CONCLUSIONS: Both polymorphisms may play an important role in the elevated TG and TRL-TG levels induced by the HC/LF diet, especially in females, thus indicating a potential dietary prevention of coronary heart disease in this Chinese cohort. PMID- 21646780 TI - Premature atherosclerosis in non-transfusion-dependent beta-thalassemia intermedia. AB - OBJECTIVES: beta-Thalassemias are inherited hemolytic anemias with a broad phenotype and an increased rate of vascular complications despite a near absence of coronary artery disease. METHODS: We investigated the presence of endothelial dysfunction and subclinical atherosclerosis in non-transfusion-dependent patients with beta-thalassemia intermedia (beta-TI) by means of flow-mediated (FMD) and flow-independent (FID) brachial artery dilatation and carotid artery intima-media thickness. RESULTS: In 20 nondiabetic young adults with beta-TI, FMD (6.6 +/- 3.7 vs. 10.3 +/- 3.1%; p = 0.002) and FID (14.0 +/- 4.7 vs. 18.0 +/- 5.6%; p = 0.02) were both lower relative to the values in 20 matched control subjects, whereas the intima-media thickness was increased (0.51 +/- 0.09 vs. 0.46 +/- 0.07 mm; p = 0.049). Fibrin generation, soluble endothelial activation markers, and proinflammatory proteins were higher in the patient group, while the plasma cholesterol level was lower. CONCLUSIONS: These findings indicate premature atherosclerosis among patients with beta-TI; this is in accord with the high incidence of noncoronary vascular episodes in beta-TI. PMID- 21646781 TI - Positron emission tomography for risk assessment of women investigated for coronary artery disease. PMID- 21646782 TI - Low risk of Clostridium difficile infections in hospitalized patients with inflammatory bowel disease in a German tertiary referral center. AB - INTRODUCTION: Many reports, mainly from the US and Canada but also a recent report from a center in Europe, have documented the increasing impact of Clostridium difficile infections in patients with inflammatory bowel disease (IBD) during the last years. To determine the prevalence of C. difficile infections in hospitalized IBD patients in a tertiary referral center in Germany, we conducted this retrospective analysis. METHODS: Data of all IBD in-patients treated due to an acute flare of their IBD at the Department of Internal Medicine I of the University of Regensburg between January 1, 2001, and June 30, 2008, were analyzed. In patients with a concomitant diagnosis of C. difficile infection, further variables such as IBD-related treatment at the time of infection or outcome were examined. RESULTS: In total, 995 in-patients with IBD were treated in this hospital [638 patients with Crohn's disease (CD), 357 with ulcerative colitis (UC)] during the study period. Of these, 279 patients with CD and 242 patients with UC were admitted with an acute flare and suffering from diarrhea and abdominal pain. Only 10 of those were diagnosed as having a concomitant infection with C. difficile. Six patients were female and the median age was 49 years (range: 15-80). Six patients with C. difficile infections suffered from UC and 4 patients from CD, all with previous colonic involvement. Eight patients used immunosuppressive therapies; only 2 patients were treated with antibiotics before infection. CONCLUSION: In contrast to recent reports from other countries, only a low percentage of hospitalized patients with acute flares of their IBD were identified as having an underlying C. difficile infection in this German tertiary referral center. However, in IBD patients with an acute flare, a concomitant C. difficile infection should be excluded, especially in patients with immunosuppressive treatment and colonic involvement of their disease. Further research is needed to evaluate if regions with different risks of C. difficile infections exist and to find out more about potential reasons for this observation. PMID- 21646783 TI - Electrophysiological assessment of the autonomic nervous system in male patients with acromegaly. AB - BACKGROUND/AIMS: We aimed to electrophysiologically evaluate the autonomic function in acromegalic patients using sympathetic skin response (SSR) as a reflection of the sympathetic sudomotor activity and RR interval variation (RRIV) as an indicator of the cardiovagal autonomic function. METHODS: The study group consisted of 18 male acromegalics, and the control group was composed of 18 age- and sex-matched healthy subjects. Participants underwent SSR and RRIV tests. Beginning latencies and amplitudes of the median and tibial SSRs were compared among the groups. The RRIV values recorded at rest and during hyperventilation were compared among the patients and controls. RESULTS: Latencies of SSRs recorded from the palms (median) and soles (tibial) of acromegalics were significantly longer than in healthy subjects (p = 0.004, p < 0.001). The amplitude of SSR recorded from the sole (tibial) was significantly decreased (p = 0.028). The RRIVs obtained from acromegalics at rest and during hyperventilation were significantly decreased compared with those of controls (p < 0.001). The RRIVs obtained from controls prolonged significantly during hyperventilation (p < 0.001); however, in the acromegaly group, hyperventilation did not cause a significant change in RRIV (p = 0.983). CONCLUSIONS: The present study suggests that an autonomic dysfunction exists in patients with acromegaly. Dysautonomia in acromegalics may be documented by means of SSR and RRIV. PMID- 21646784 TI - Simultanagnosia. PMID- 21646785 TI - A novel end-to-side anastomosis technique for hepatic rearterialization in rat orthotopic liver transplantation to accommodate size mismatches between vessels. AB - BACKGROUND/AIMS: We present our modification of a sutured arterial anastomosis in orthotopic rat liver transplantation as well as a literature survey and analysis of the existing techniques of rearterialization with regard to technical difficulties and potential limitations. METHODS: The donor common hepatic artery (CHA) was anastomosed to the enlarged lumen of the recipient proper hepatic artery (PHA), tailored to match the size of the donor CHA, with an end-to-side interrupted suture technique. Vascular patency of hepatic rearterialization was assessed both intraoperatively and at the time the liver grafts were harvested (postoperative days 2 and 28). The effect of arterialization on hepatic morphology was confirmed by histological examination and compared to nonarterialized rat orthotopic liver transplantation. RESULTS: The CHAs had a significantly larger diameter (up to 3-fold) compared to the PHAs, which represents a considerable size mismatch. The anastomosis procedure including the size adaptation required 15-25 min. All anastomoses were patent immediately, 5 min after rearterialization and at both harvest time points. The liver lobular architecture was intact in the rearterialized group, whereas a moderate degree of bile duct proliferation and portal/lobular lymphocytic infiltration were observed in the nonarterialized group. CONCLUSION: The new technique is a time-consuming and microsurgically challenging but universally applicable and robust procedure accommodating even a substantial mismatch in vessel diameter. PMID- 21646786 TI - A rapid quantitative real-time PCR-based DNA quantification assay coupled with species--assignment capabilities for two hybridizing Macaca species. AB - Regional populations of rhesus and long-tailed macaques exhibit fundamental differences in mitochondrial DNA, short tandem repeat and single nucleotide polymorphism variation between mainland and insular Southeast Asian populations. Some studies have revealed genetic admixture between these species due to natural hybridization and human-assisted intercrosses. A quantitative real-time PCR (qPCR) assay was developed to efficiently determine the species of origin of a macaque biological sample, and to quantify the species-specific template DNA. Prior knowledge of species identity and DNA concentrations are crucial for maintaining cost-effective methods and accurate DNA analysis. DNA from 109 regionally representative rhesus and long-tailed macaques was qPCR amplified to determine the species and template quantities. Of the 19 Vietnamese long-tailed macaques, 3 samples were discovered to be hybrids. PMID- 21646787 TI - Eosinophils in allergy and related diseases. Proceedings of a Workshop, Tokyo, Japan, June 19, 2010. Preface. PMID- 21646788 TI - Qualitative evaluation of adjuvant activities and its application to Th2/17 diseases. AB - Dendritic cells (DCs) are antigen-presenting cells specialized to activate naive T lymphocytes and initiate primary immune responses. The different classes of specific immune responses are driven by the biased development of antigen specific helper T cell subsets - that is, Th1, Th2, and Th17 cells - that activate different components of cellular and humoral immunity. DCs reside in an immature state in many nonlymphoid tissues such as the skin or airway mucosa which are highly exposed to allergens, pathogens, and chemicals. T cell receptor stimulation with costimulation allows naive Th cells to develop into effector cells, normally accompanied by high-level expression of selective sets of cytokines. The balance of these cytokines and the resulting class of immune responses depend on the conditions under which DCs are primed. Immunomodulators such as lipopolysaccharides/forskolin/curdlan change the nature of DCs to induce Th1/Th2/Th17 cells thereby designated Th1/Th2/Th17 adjuvants. We have recently found that such activities can be scrutinized by using mixed lymphocyte reaction, cAMP, and differential expression of Notch ligand isoforms. Application of these methods for the analyses of atopic dermatitis and experimental autoimmune encephalomyelitis will be discussed. PMID- 21646789 TI - Role of prostaglandin D2 receptor CRTH2 in sustained eosinophil accumulation in the airways of mice with chronic asthma. AB - The prostaglandin D(2) (PGD(2))/CRTH2 pathway is important for eosinophil trafficking in vitro; however, genetic deficiency of CRTH2 does not suppress in vivo eosinophilic airway inflammation in acute models of asthma, and the role of CRTH2 in the pathogenesis of asthma is still ambiguous. Therefore, in the present study we explored whether the PGD(2)/CRTH2 pathway could affect the phenotypes of chronic asthma. Either CRTH2-deficient (CRTH2-/-) or wild-type mice were sensitized and exposed to ovalbumin (OVA) for 3 days (acute model) or 6 weeks (chronic model). While the magnitude of the acute eosinophilic inflammation was equivalent between CRTH2-/- and wild-type mice, the number of inflammatory cells and eosinophils in bronchoalveolar lavage fluid after chronic OVA exposure was significantly reduced in CRTH2-/- mice (18.0 +/- 2.6 * 10(4) cells and 2.0 +/- 0.5 * 10(4) cells) compared to wild-type mice (27.9 +/- 2.5 * 10(4) cells and 6.8 +/- 1.1 * 10(4) cells, p < 0.001). On the contrary, no difference was observed between CRTH2-/- and wild-type mice in terms of airway hyperresponsiveness or remodeling (goblet cell hyperplasia) in the chronic model of asthma. In conclusion, CRTH2 that mediates PGD(2) activity is essential for sustained eosinophilic inflammation in the airways, and its antagonists could exert an anti inflammatory effect in chronic asthma. PMID- 21646790 TI - Expression and effects of IL-33 and ST2 in allergic bronchial asthma: IL-33 induces eotaxin production in lung fibroblasts. AB - BACKGROUND: Interleukin (IL)-33, a new member of the IL-1 cytokine family, has been recognized as a key cytokine that enhances T helper 2-balanced immune regulation through its receptor ST2; however, the function and relationship of the IL-33 and ST2 pathways in bronchial asthma are still unclear. We investigated the cellular origin and regulation of IL-33 and ST2 in allergic bronchial asthma in vivo and in vitro. METHODS: BALB/c mice were sensitized by intraperitoneal injections of ovalbumin (OVA) with alum. Mice were exposed to aerosolized 1% OVA for 30 min a day for 7 days. These mice were then challenged with aerosolized 1% OVA 2 days after the last day of exposure. After the OVA challenge, the mice were sacrificed and their lung tissues were obtained. Mouse lung fibroblasts were cultured and treated with IL-33 or IL-13. RESULTS: The levels of IL-33 mRNA and IL-33 protein in lung tissue increased after the OVA challenge. Most IL-33 expressing cells were CD11c+ cells and epithelial cells, and many ST2-expressing cells were stained lung fibroblasts and inflammatory cells. IL-33 induced eotaxin/CCL11 production in lung fibroblasts. IL-33 and IL-13 synergistically induced eotaxin expression. CONCLUSIONS: IL-33 may contribute to the induction and maintenance of eosinophilic inflammation in the airways by acting on lung fibroblasts. IL-33 and ST2 may play important roles in allergic bronchial asthma. PMID- 21646791 TI - T cell subsets related with a sex difference in IL-5 production. AB - BACKGROUND: Before puberty, the prevalence and severity of asthma are higher in boys than in girls, but this pattern is reversed after puberty. The underlying mechanisms of these gender differences in asthma are not fully understood. Using murine models of allergic asthma, a sex difference in Th2 cytokine production has been suggested to contribute to the gender differences in asthma. Therefore, we determined which subsets of T cells are involved in the sex difference in Th2 cytokine production. METHODS: Splenocytes from wild-type mice and CD4+ T cell-, CD8+ T cell-, and iNKT cell-deficient mice were stimulated with anti-CD3/CD28 antibodies for 3 days, and the concentrations of IL-4, IL-5, IL-13, and IFN-gamma in the cultures were measured by ELISA. RESULTS: IL-5, but not IL-4 and IL-13, concentrations in culture derived from female wild-type mice were significantly higher than those in male wild-type mice. The sex difference in IL-5 concentrations was not observed in the cultures of splenocytes from CD4+ and CD8+ T cell-deficient mice. The disappearance of the sex differences in CD4+ and CD8+ T cell-deficient mice was attributable to a decrease in IL-5 concentration in female mice and an increase in IL-5 concentration in male mice. In iNKT cell deficient mice, the sex difference was still observed. There was no significant difference between the sexes in any type of mice with respect to IFN-gamma production. CONCLUSIONS: There was a sex difference in IL-5 production by splenocytes stimulated by TCR activation. The difference might be attributable to sex differences in CD4+ and CD8+ T cell functions. PMID- 21646792 TI - Peripheral blood mononuclear cells from patients with bronchial asthma show impaired innate immune responses to rhinovirus in vitro. AB - BACKGROUND: Asthmatic patients have a higher susceptibility to rhinovirus (RV) infection, and impaired IFN-beta and IFN-lambda production has been demonstrated in bronchial epithelial cells from asthmatic adults upon exposure to RV. However, the mechanisms underlying the increased susceptibility of asthmatic patients to RV infection remain poorly understood. The present study aimed to elucidate the characteristics of the immune responses of asthmatic patients' peripheral blood mononuclear cells (PBMCs) to RV exposure. METHODS: PBMCs obtained from 3 different age groups (2-6 years: young-children group; 7-19 years: youth group; >=20 years: adult group) of asthmatic patients and nonasthmatic control subjects were stimulated with RV-14 for 72 h. Healthy adults with a history of childhood asthma were also enrolled. The concentrations of IFN-alpha, IL-6, TNF-alpha, IL 10, and soluble Fas ligand (sFasL) in the culture supernatants were measured by ELISA. RESULTS: When compared with age-matched control subjects, IFN-alpha production was significantly lower in the asthmatic youth group. IL-6, TNF-alpha, IL-10, and sFasL productions were significantly lower in both the asthmatic youth group and the adult group. Such impaired responses were not found in healthy adults with a history of childhood asthma. No significantly different responses were found between the asthmatics and controls in the young-children group, whereas young asthmatic children with persistent wheeze during a 2-year follow-up showed significantly lower IL-10 production than those without wheeze. CONCLUSIONS: These results imply the involvement of impaired production of both IFN-alpha and inflammatory cytokines seen in asthmatic patients' PBMCs upon exposure to RV in the higher susceptibility of those patients to RV infection. PMID- 21646793 TI - Human eosinophils produce and release a novel chemokine, CCL23, in vitro. AB - BACKGROUND: CCL23 (MPIF1/CK-BETA-8) is a novel CC chemokine that plays important roles in the inhibition of myeloid progenitor cell development, the selective recruitment of resting T lymphocytes and monocytes, and the potentiation of VEGF induced proliferation and migration of human endothelial cells. Since eosinophils participate in the pathogenesis of airway remodeling, we examined CCL23 production and release by human eosinophils in vitro. METHODS: Using Ficoll and antibody-coated immunomagnetic beads, eosinophils and other blood cells were purified from peripheral blood samples obtained from normal subjects and mildly allergic patients. Eosinophils were cultured in the presence of 10 ng/ml granulocyte-macrophage colony-stimulating factor (GM-CSF), 10 ng/ml IL-5, 100 ng/ml IFN-gamma, 100 ng/ml IFN-alpha, or immobilized secretory IgA (sIgA). Total mRNA was extracted after 6 h of culture, and mRNA expression was measured using a microarray and RT-PCR. The CCL23 concentrations in the supernatants and cell lysates after 24 and 48 h of culture were measured by ELISA. RESULTS: CCL23 mRNAs (both CK-beta8-1 and CK-beta8) were constitutively expressed in fresh eosinophils, and their expression levels were higher than in other types of blood cells. CCL23 mRNAs were significantly increased by stimulation with GM-CSF and IL 5 and slightly by IFN-alpha and immobilized sIgA. Fresh eosinophils contained trace amounts of CCL23 protein. CCL23 was significantly released into the supernatant when the eosinophils were stimulated with GM-CSF or IL-5 but not with IFN-gamma or immobilized sIgA. CONCLUSION: Our data suggest that eosinophils produce and release CCL23 and may be involved in some in vivo physiological and pathological conditions. PMID- 21646794 TI - Eosinophilic gastrointestinal disorders in infants: a Japanese case series. AB - BACKGROUND: Eosinophilic gastrointestinal disorders (EGIDs) are disorders characterized by primary eosinophil inflammation in the gastrointestinal tract. There are a small number of reports of eosinophil infiltration in gastrointestinal tracts presenting as EGIDs in infants. In this study, we present Japanese cases of EGIDs in infants. METHODS: Five patients diagnosed with or strongly suspected to have EGIDs in our hospital from 2008 to 2010 were reviewed. Radiographic contrast enema examinations and/or endoscopies were performed in 4 and 3 patients, respectively. RESULTS: There were patients with eosinophilic colitis (1 suspected and 2 biopsy-proven), a patient who was suspected of having allergic eosinophilic enterocolitis, and a patient with eosinophilic gastroenteritis associated with pediatric hypereosinophilic syndrome. CONCLUSIONS: The causes and clinical findings of patients with intestinal eosinophil inflammation vary. Therefore, deliberate examination and observation are important for patients with infantile EGID. PMID- 21646795 TI - The proportion of regulatory T cells in the peripheral blood reflects the relapse or remission status of patients with Churg-Strauss syndrome. AB - BACKGROUND: Churg-Strauss syndrome (CSS) is a rare, systemic, necrotizing vasculitis that develops in some asthma patients. We previously reported that maintenance of the proportion of type 1 regulatory T (Tr1) cells in patients with chronic eosinophilic pneumonia and asthma might inhibit the development of CSS through the action of cytokines, such as interleukin (IL)-10 and IL-2, produced by Tr1 and responder T cells. We also reported that IL-17-producing CD4+ helper T cells (Th17 cells) are involved in the pathogenesis of CSS because a higher proportion of Th17 cells was observed in CSS patients during relapses than during remissions. However, few studies have addressed the role of both Tr1 cells and Th17 cells in the status of CSS. METHODS: We recruited 40 patients (25 in remission and 15 in relapse) for participation in this study. CSS was diagnosed on the basis of American College of Rheumatology criteria. Remission was defined as the absence of any clinical symptoms of active vasculitis. Tr1 cells were defined as CD4+CD25+ T cells that predominantly produce IL-10 when costimulated with phorbol myristate acetate (PMA) and ionomycin. Naturally occurring Treg (nTreg) cells were defined as CD4+CD25+ T cells that expressed Forkhead box P3 (FOXP3) and cytotoxic T-lymphocyte antigen 4 (CTLA-4). Th17 cells were identified as CD4+ T cells that mainly produced IL-17 and IL-22. Peripheral blood mononuclear cells (PBMCs) obtained from the subjects were costimulated with PMA and ionomycin, and intracellular cytokines were detected after fixing and permeabilizing the cells. Indoleamine 2,3-dioxygenase (IDO) expression was measured in PBMCs that had been treated with IFN-gamma and then stimulated overnight with lipopolysaccharide (LPS) or lipopeptide Pam3CSK. RESULTS: Lower expression of CTLA-4 was observed on the surface of CD4+CD25+ T cells obtained from patients with relapsed CSS versus patients in remission. Both FOXP3 expressing nTreg cells and IL-10-producing Tr1 cells were detected in a lower proportion in patients with a relapse compared to patients in remission, but the proportion of CD4+ T cells producing IL-17 was higher during relapse than during remission. In addition, the proportion of CD4+ T cells that produced IL-25, which promotes Th2 inflammation, was also higher in the relapsed patients. We observed a lower percentage of CD14+ monocytes expressing both TLR2 and TLR4 obtained from patients with a relapse of CSS versus patients in remission. Stimulation of CD14+ monocytes with LPS or Pam3CSK reduced IDO expression by the cells from patients with relapsed CSS. The level of IDO expression was positively correlated with the proportion of Tr1 cells in the peripheral blood and inversely correlated with the percentage of Th17 cells. CONCLUSION: CSS relapse may be linked to increased numbers of CD4+ T cells producing IL-25, which promotes Th2 inflammation, and to a decline in the Tr1 cell subpopulation resulting from lower IDO expression in monocytes. Thus, the proportions of Tr1 cells and Th17 cells reflect the status of CSS. PMID- 21646796 TI - Zinc finger protein, multitype 1, suppresses human Th2 development via downregulation of IL-4. AB - BACKGROUND: Among several GATA family transcription factor-associating proteins, zinc finger protein, multitype 1 (ZFPM1), at least that of murine origin, has been shown to modulate the activity of GATA-3. However, the functional role of human ZFPM1 in the immune system has not been elucidated. Therefore, we here investigated the contribution of ZFPM1 to human Th1/Th2 differentiation. METHODS: The cDNA of ZFPM1 was cloned and introduced into human cord blood CD4+ T cells by a lentiviral transduction system. Then, the expression of IL-4 and IFN-gamma mRNA was determined by quantitative real-time RT-PCR. The effect of ZFPM1 on the promoter activity of IL-4 and IFN-gamma in Jurkat cells was also investigated. RESULTS: Stimulation-induced expression of IL-4 and IFN-gamma in human CD4+ T cells was suppressed and enhanced, respectively, by the introduction of ZFPM1. The transcriptional activity of IL-4 was also diminished by ZFPM1, whereas that of IFN-gamma was not affected. CONCLUSION: ZFPM1 that facilitates human Th1 differentiation via the downregulation of IL-4 is a potential target for the treatment of allergic diseases. PMID- 21646797 TI - Peroxisome proliferator-activated receptor-gamma is expressed in eosinophils in nasal polyps. AB - BACKGROUND: Peroxisome proliferator-activated receptor-gamma (PPARgamma) is a member of the nuclear receptors, which regulate fatty acid metabolites. One of the natural ligands for PPARgamma is 15-deoxy-Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)), a major metabolite of prostaglandin D(2) (PGD(2)). Recently, PPARgamma has been shown to play an important role in anti-inflammatory reactions, including within-airway allergic diseases, in a mouse model. Our aim was to clarify the expression and localization of PPARgamma and PGD(2) synthase, which produces ligands of PPARgamma, in nasal polyps by immunohistochemical analysis. METHODS: Nasal polyps of chronic rhinosinusitis patients (6 asthmatic patients and 6 nonasthmatic patients) were obtained during surgery. May-Grunwald Giemsa staining was performed to evaluate the eosinophil infiltration of the polyps. To identify the cells expressing PPARgamma protein and PGD(2) synthase, double immunostaining was performed using anti-PPARgamma antibody, monoclonal antileukocyte antibodies, and PGD(2) synthase antibody. RESULTS: The number of eosinophils and the number of PPARgamma-positive cells in the nasal polyps of the asthmatic patients were significantly higher than those in the nonasthmatic patients. PPARgamma was expressed on eosinophils and T cells of the infiltrating cells in the nasal polyps. PGD(2) synthase was also expressed on PPARgamma positive cells. CONCLUSION: PPARgamma is involved in nasal polyposis pathogenesis, acting on eosinophils and T cells. PMID- 21646798 TI - The in vitro effects of advanced glycation end products on basophil functions. AB - BACKGROUND: Basophils are thought to play pivotal roles in the pathogenesis of allergic reactions, but their roles in inflammation associated with systemic abnormalities such as metabolic disorders remain largely unknown. Advanced glycation end products (AGEs) are potentially important substances produced in high-glucose disease conditions. In this in vitro study, we investigated whether the biological functions of human basophils can be influenced by AGEs. METHODS: We analyzed the effects of AGEs on various functions and markers of human basophils, including CD11b expression, apoptosis, degranulation, and cytokine production. RESULTS: Flow cytometric analysis indicated that the level of the receptor for AGEs (RAGE) on the surface of freshly isolated basophils was very low but was clearly upregulated by IL-3. Apoptosis of basophils was induced by high concentrations of glycated albumin. Although glycated albumin failed to affect the level of surface CD11b expression or to trigger degranulation or production of IL-4 and IL-13 in basophils, it dose-dependently induced IL-6 and IL-8 secretion. CONCLUSIONS: AGEs seem to act on human basophils; they suppress the cells' longevity but elicit secretion of inflammatory cytokines. Through these biological changes, basophils might play some roles in inflammatory conditions associated with metabolic disorders presenting elevated levels of AGEs. PMID- 21646799 TI - STAT4 is required for IFN-beta-induced MCP-1 mRNA expression in murine mast cells. AB - BACKGROUND: Mast cells are immunocompetent cells that are found in almost all tissues and function as sentinels of immune responses. Recently, it has been shown that mast cells play significant roles in innate immune responses. However, it is still largely unknown whether signal transducers and activators of transcription 4 (STAT4), one of the STAT proteins under type I IFN signaling, is involved in type I IFN-mediated gene expression in mast cells. METHODS: We investigated the role of STAT4 in IFN-beta-induced gene expression in mast cells by using STAT4-deficient (STAT4(-/-)) bone marrow-derived mast cells (BMMCs). RESULTS: STAT4 was expressed in BMMCs and activated in response to IFN-beta but not to IL-12 or IL-23. The development of BMMCs as well as IgE-induced degranulation of BMMCs was normal in STAT4(-/-) mice. On the other hand, while IFN-beta-induced mRNA expression of interferon-induced protein with tetratricopeptide repeats 1 (IFIT-1), protein kinase interferon-inducible double stranded RNA dependent (PKR), and myxovirus resistance 1 (Mx1) was similar between STAT4(-/-) BMMCs and wild-type (WT) BMMCs, IFN-beta-induced MCP-1 mRNA expression was severely diminished in STAT4(-/-) BMMCs as compared with WT BMMCs. CONCLUSIONS: STAT4 plays an essential role in IFN-beta-induced MCP-1 mRNA expression in mast cells. PMID- 21646801 TI - Inhibition of eosinophil activation mediated by a Toll-like receptor 7 ligand with a combination of procaterol and budesonide. AB - BACKGROUND: Viral respiratory tract infections play an important role in the inception and exacerbation of asthma. Eosinophils, major effector cells in asthma, often accumulate in the airways during viral infections and are possibly activated by respiratory RNA viruses through Toll-like receptor (TLR) 7. We investigated the effect of a beta(2)-agonist, i.e. procaterol, and a corticosteroid, i.e. budesonide, that are commonly used for viral-induced asthma, on TLR7 ligand-induced activation of eosinophils in vitro. METHODS: Purified peripheral blood eosinophils were incubated with procaterol and/or budesonide and stimulated with a TLR7 ligand, i.e. R-837. Expression of CD11b was analyzed by flow cytometry. Superoxide generation was measured via the cytochrome C reduction method. IL-8 in the supernatants was assayed by ELISA. RESULTS: Although procaterol or budesonide alone did not inhibit R-837-induced CD11b expression, combinations of the 2 drugs significantly inhibited CD11b. Likewise, the combinations significantly inhibited O(2)(-) generation at low concentrations. Budesonide significantly inhibited R-837-induced IL-8 production in a concentration-dependent manner, and procaterol potentiated inhibition by budesonide although single-agent procaterol had no effect. CONCLUSION: A combination of procaterol and budesonide inhibits the TLR7-mediated effector function of eosinophils, indicating their possible anti-inflammatory effect for virus-induced asthma. PMID- 21646800 TI - Differential effects of corticosteroids on serum eosinophil cationic protein and cytokine production in rhinovirus- and respiratory syncytial virus-induced acute exacerbation of childhood asthma. AB - BACKGROUND: Little information is available on eosinophil activation and the cytokine profile in virus-induced acute exacerbation of bronchial asthma; therefore, we examined the effects of treatments that included systemic corticosteroids on serum eosinophil cationic protein (ECP) and 17 cytokines/chemokines in rhinovirus- and respiratory syncytial (RS) virus-induced acute exacerbation of childhood asthma. METHODS: We measured the peripheral eosinophil count, as well as the serum levels of ECP and 17 types of cytokines/chemokines (IL-1beta, 2, 4, 5, 6, 7, 8, 10, 12, 13, and 17 and IFN gamma, TNF-alpha, GM-CSF, G-CSF, MCP-1, and MIP-1beta), using a multiplex bead based assay in 21 cases of rhinovirus- and 12 cases of RS virus-induced acute exacerbation of childhood asthma and 13 controls. We also compared the clinical data and the effects of systemic corticosteroids on these responses between rhinovirus and RS virus groups. RESULTS: The serum levels of ECP, IL-5, and IL-6 were significantly elevated in patients with rhinovirus-induced acute exacerbation of asthma compared with controls, while serum IL-1beta and IFN-gamma were significantly lower in patients with rhinovirus-induced acute exacerbation of asthma than in controls. On the other hand, in RS virus-induced acute exacerbation of asthma, only the peripheral eosinophil count was significantly decreased compared with that in rhinovirus-induced acute exacerbation of asthma and controls. Furthermore, the serum levels of ECP, IL-5, and IL-6 in rhinovirus induced acute exacerbation of asthma and levels of G-CSF in RS virus-induced acute exacerbation of asthma were significantly reduced after treatments that included systemic corticosteroids, respectively. CONCLUSION: These results suggest that the effects of systemic corticosteroids on serum ECP and the cytokine profile are different between rhinovirus- and RS virus-induced acute exacerbation of childhood asthma. PMID- 21646802 TI - Effects of a cysteinyl leukotriene dual 1/2 receptor antagonist on antigen induced airway hypersensitivity and airway inflammation in a guinea pig asthma model. AB - BACKGROUND: Little is known about the role of the cysteinyl leukotriene (cysLT) 2 receptor in the pathophysiology of asthma. The aim of this study is to investigate the effects of a cysLT1 receptor antagonist (montelukast) and a dual cysLT1/2 receptor antagonist (BAY-u9773) on airway hypersensitivity and airway inflammation induced by antigen challenge in ovalbumin (OVA)-sensitized guinea pigs. METHODS: Male Hartley guinea pigs sensitized with OVA were intraperitoneally administered 0.1, 1, or 10 mg/kg of montelukast or 0.1 mg/kg of BAY-u9773 and then challenged with inhaled OVA. Airway reactivity to acetylcholine, inflammatory cells in bronchoalveolar lavage (BAL) fluid, and eosinophil infiltration in airway walls after OVA challenge were evaluated. RESULTS: Pretreatment with 1 or 10 mg/kg, but not 0.1 mg/kg, of montelukast significantly suppressed airway hypersensitivity and eosinophil infiltration into the BAL fluid. Moreover, 0.1 mg/kg of BAY-u9773 significantly suppressed the development of these markers. The suppressive effects of BAY-u9773, although not significantly different, trended toward being greater than those of montelukast. Although all of the doses of montelukast tested and 0.1 mg/kg of BAY-u9773 significantly suppressed eosinophil infiltration in airway walls, the suppressive effect of BAY-u9773 was significantly greater than that of 0.1 mg/kg of montelukast. CONCLUSION: Signaling may contribute to the pathophysiology of asthma via the cysLT1/2 receptor. PMID- 21646803 TI - Utility of the peripheral blood basophil histamine release test in the diagnosis of hen's egg, cow's milk, and wheat allergy in children. AB - BACKGROUND: The diagnosis of food allergy (FA) is made by oral food challenge tests (OFCs) that occasionally produce serious symptoms in patients; therefore, whether to perform OFCs should be carefully considered. The utility of the histamine release test (HRT) in the diagnosis of childhood FA has not been fully examined. METHODS: Sixty-four subjects with suspected hen's egg allergy, cow's milk allergy (CMA), and wheat allergy (WA) were enrolled. The diagnosis of FA was made based on the outcomes of OFCs or a convincing history of symptoms after food ingestion within 6 months before or after sample collection. HRT was performed using an HRT Shionogi kit. The threshold of histamine release (HRT threshold), which was defined as the minimum concentration of food antigen to induce a 10% net histamine release, was analyzed in association with FA diagnosis. RESULTS: Receiver operating characteristic analysis showed that the HRT threshold was useful in the diagnosis of heated egg allergy (HEA), raw egg allergy (REA), CMA, and WA. We were able to determine the cutoff value for the HRT threshold in relation to outcomes of OFCs. The cutoff value was 6 ng/ml of egg white antigen in HEA and REA (p < 0.01), 40 ng/ml of milk antigen in CMA (p < 0.01), and 500 ng/ml of wheat antigen in WA (p < 0.05). The efficiency was 70.3% for HEA, 78.0% for REA, 77.6% for CMA, and 70.7% for WA. CONCLUSIONS: We conclude that the HRT threshold measurement for egg white, milk, and wheat antigen is related to outcomes of OFCs and is useful in determining when OFCs should be performed. PMID- 21646804 TI - Protease-mediated house dust mite allergen-induced reactive oxygen species production by neutrophils. AB - A growing body of evidence indicates that neutrophils may play an important role in the pathogenesis of asthma. However, the involvement of the house dust mite (HDM) in neutrophil activation associated with the pathogenesis of asthma is not fully understood yet. To address this situation, we harvested neutrophils isolated from 15 HDM-sensitized asthmatic subjects and 18 HDM-sensitized nonasthmatic subjects and measured the amounts of neutrophil reactive oxygen species (ROS) production in response to the major HDM allergens Der-f and Der-f1. Der-f and Der-f1 significantly increased ROS production in neutrophils isolated from asthmatic subjects versus nonasthmatic subjects. To assess the involvement of Der-f-specific IgE antibodies binding to their receptors in HDM allergen induced ROS production, we examined whether neutrophils produce ROS by cross linking of cell-bound IgE antibodies with anti-IgE. Treatment with anti-IgE antibodies did not induce ROS production by neutrophils isolated from 6 asthmatic subjects. On the other hand, pretreatment of Der-f with E-64, a cysteine protease inhibitor, eliminated Der-f-induced ROS production. These results suggest that HDM-allergen exposure may result in greater production of ROS in asthmatic patients and may be involved in the pathogenesis of asthma. PMID- 21646805 TI - Comparative analysis of steroid sensitivity of T helper cells in vitro and in vivo. AB - BACKGROUND: Glucocorticoid (GC) action on asthma has been partly explained by the inhibition of T cell activation. We analyzed the steroid sensitivity of ovalbumin (OVA) reactive helper T (Th) cell clones both in vitro and in vivo. METHOD: For in vitro experiments, Th clones were cultured with antigen-presenting cells, OVA, and various concentrations of dexamethasone (DEX). The proliferative response of each Th clone was measured by (3)H-thymidine uptake. For in vivo experiments, unprimed BALB/c mice were transferred with Th clones, challenged with OVA, and administered DEX subcutaneously. The number of infiltrating cells in bronchoalveolar lavage fluid (BALF) was measured. RESULTS: Six Th clones were classified as steroid-sensitive or steroid-resistant clones in terms of the effects of GC on the proliferative responses analyzedin vitro. Airway infiltration of eosinophils and lymphocytes of mice transferred with steroid sensitive clones were effectively inhibited by the administration of DEX. In contrast, those of mice transferred with steroid-resistant clones were not significantly inhibited by DEX; the number of eosinophils in the BALF of mice transferred with 1 steroid-resistant clone, i.e. T5-1, was only partially reduced. CONCLUSION: The steroid sensitivity of Th clones measured in vitro was consistent with that of an adoptively transferred asthma model measuredin vivo. Steroid-sensitive and resistant asthma models seem valuable for understanding the mechanisms of steroid resistance in severe asthma. PMID- 21646806 TI - An 8-year-old boy with hypereosinophilic syndrome. AB - Hypereosinophilic syndrome (HES) is a heterogeneous group of uncommon disorders characterized by the presence of marked peripheral blood eosinophilia and tissue eosinophilia, resulting in a wide variety of clinical manifestations. We present the case of an 8-year-old boy with HES. He complained of recurrent abdominal pain, general fatigue, and diarrhea. Laboratory data showed marked eosinophilia, elevated total IgE with positive specific IgE antibodies to common inhalant and food allergens, and elevated serum CCL17/TARC. A chest CT scan revealed central bronchiectasis, bronchial wall thickening, a mosaic attenuation pattern, and multiple small nodules in lung parenchyma; abdominal CT showed a thickened bladder wall. Gastrointestinal endoscopy revealed scarring in the gastric mucosa and mucosal erosion in the duodenum. Immunohistochemical examination demonstrated numerous eosinophil infiltrations with extensive extracellular eosinophil major basic protein deposition in the gastric mucosa. Only high-dose oral steroid was effective and cyclosporine appeared to have a steroid-sparing effect. HES is extraordinary rare in children and the long-term prognosis in pediatric HES is not well known. Comprehensive diagnostic procedures are vital for the early detection and management of complications in pediatric HES. PMID- 21646807 TI - Human eosinophil innate response to Alternaria fungus through protease-activated receptor-2. AB - Eosinophils are multifunctional leukocytes implicated in the pathogenesis of allergic diseases. An association between eosinophilic inflammation and infection or colonization by fungi has also been long recognized. However, the mechanisms underlying how eosinophils are activated and how they release proinflammatory and immunomodulatory mediators such as major basic protein (MBP) and eosinophil derived neurotoxin remain largely unknown. We used a fungus, i.e. Alternaria, as a model microbe relevant to human asthma and chronic rhinosinusitis (CRS) to investigate the molecular mechanisms involved in the immune recognition of ubiquitous environmental allergens. Human eosinophils are activated by live Alternaria alternata organisms, release their granule proteins, and kill the fungi. Eosinophils, but not neutrophils, respond to secreted products from A. alternata. We found that eosinophils are equipped with innate cellular activation machinery that responds to an extracellular aspartate protease secreted by Alternaria. Aspartate protease activation of protease-activated receptor (PAR)-2 probably involves a novel mechanism different from that for serine protease activation of PAR-2. Thus, human eosinophils may recognize certain danger signals or virulence factors produced by fungi and provoke inflammatory responses against these organisms. Dysregulation of such an innate immune mechanism may be involved in the pathophysiology of certain human inflammatory diseases, including asthma and CRS. PMID- 21646808 TI - Role of the arylhydrocarbon receptor in lung disease. AB - Ubiquitous environmental contaminants such as dioxins have long been implicated in cellular toxicity, but only recently have various biological effects been linked to immune regulation. These plentiful noxious agents exert their effects through the arylhydrocarbon receptor (AhR) recognized as a ligand-activated transcription factor. AhR activation mediates gene alteration, cell-cell adhesion interaction, cytokine expression, and mucin production, which are involved in the induction of cancer or inflammation. We have reported that human bronchial epithelial cells express AhR, and AhR activation induces mucin production through reactive oxygen species. This review discusses the role of AhR in lung disease, focusing in particular on airway epithelial cells. In addition, although it is not yet clear how the activation of AhR modifies carcinogenesis or airway inflammation, we mention a potent therapeutic target for AhR activation in the prevention/treatment of lung cancer, allergic asthma, and chronic obstructive pulmonary disease. PMID- 21646809 TI - Cockroach induces inflammatory responses through protease-dependent pathways. AB - Exposure to cockroaches is a major risk factor for asthma. Products from cockroaches may contain proteases and ligands for pattern recognition receptors. These molecules may activate airway inflammatory cells, such as eosinophils, that are involved in asthma. Among inner-city children, cockroach allergens play an especially important role in increasing asthma morbidity. The molecular mechanism for this association between cockroach exposure and asthma is not fully understood. Enzymatic activities from cockroaches activate inflammatory cells in the airways and may also exacerbate certain human airway diseases, such as asthma. We recently reported that cockroach extracts contain pepstatin A sensitive proteases that activate PAR-2 and induce activation and degranulation of human eosinophils. This review focuses on the effects of cockroach on various inflammatory cells, including eosinophils, epithelial cells, fibroblasts, dendritic cells, and T cells, in allergic reactions. PMID- 21646810 TI - Thioredoxin in allergic inflammation. AB - Thioredoxin (TRX) is a redox-active protein that regulates reactive oxidative metabolism and plays a crucial role in the antioxidant system in regulating the reduction/oxidation balance by scavenging reactive oxygen species, which is implicated in the mechanism of asthma. As for the mechanisms by which TRX exerts its beneficial effects, some studies have shown that TRX suppresses allergic inflammation in animal models of asthma. Recently, we reported that TRX directly modulated the chemotaxis of eosinophils, which have been shown to play a pivotal role in the mechanism of allergic airway inflammation, in the absence of T helper (Th)1 or Th2 cytokines. Further, serum TRX levels in patients with asthma were significantly increased in patients with attacks compared with those in the asymptomatic period. This review focuses on TRX in allergic reactions and discusses the physiological role of TRX. PMID- 21646812 TI - Nutrigenetics and prostate cancer: 2011 and beyond. AB - BACKGROUND/AIMS: Prostate cancer runs in families and shows a clear dietary involvement. Until recently, the key risk gene(s) have proved elusive. We summarise current understandings of nutrient-gene interactions in prostate cancer risk and progression. METHODS: A MEDLINE-based literature search was conducted. RESULTS: Hypothesis-directed candidate gene approaches provide plausible, albeit statistically weak, nutrient-gene interactions. These are based on early understandings of factors likely to impact on carcinogenesis, including both nutrient and genetic effects on androgen biosynthesis and action, xenobiotic metabolism, DNA damage and DNA repair. Non-hypothesis-directed genome-wide association studies provide much stronger evidence for other genes, not hitherto suspected for involvement. Although only a few of these have been formally tested for dietary associations in well-designed epidemiologic studies, the nature of many of the genes suggests that their activity may be regulated by nutrients. These effects may not only be relevant to prostate cancer susceptibility, but also to disease progression. CONCLUSIONS: It will be important to move beyond studying single nucleotide polymorphisms, into more complex chromosomal rearrangements and to epigenetic changes. For future progress, large international cohorts will not only need to provide proof of individual nutrient gene interactions, but also to relate these to more complex nutrient-gene-gene interactions, as parts of pathways. Bioinformatics and biostatistics will be increasingly important tools in nutrigenetic studies beyond 2011. PMID- 21646813 TI - Interactions between dietary fat intake and FASN genetic variation influence LDL peak particle diameter. AB - BACKGROUND: The small, dense LDL phenotype is associated with an increased cardiovascular disease risk. A genome-wide scan performed on the Quebec Family Study (QFS) revealed a quantitative trait locus for LDL peak particle diameter (LDL-PPD) on the 17q21 region. A positional candidate gene - the fatty acid synthase gene (FASN) - encodes a key enzyme in the biogenesis of membrane lipids. FASN may play a role in the regulation of feeding and may be a potential therapeutic target for obesity and insulin resistance. METHODS: Analyses were performed on 592 subjects of the QFS. Dietary fat was estimated by a 3-day food record. LDL-PPD was measured by gradient gel electrophoresis on polyacrylamide gradient gels. RESULTS: Five single nucleotide polymorphisms were genotyped in FASN gene. FASN rs4246444 was associated with LDL-PPD, but only when fat intake was taken into account (p = 0.001). High and low lipid consumers were defined using a cutoff of 35% of dietary fat intake. Carriers of the variant allele showed smaller LDL-PPD only when consuming a high amount of fat. This association remained significant after adjustments for age, sex, body mass index and plasma triglyceride levels. CONCLUSION: The results suggest that dietary fat intake may modify the effect of the FASN rs4246444 polymorphism on LDL-PPD. PMID- 21646814 TI - Higher serum levels of free K plus lambda immunoglobulin light chains ameliorate survival of hemodialysis patients. AB - BACKGROUND/AIMS: Impaired immune function is common in patients with chronic renal failure. Now, we determined whether serum levels of free immunoglobulin light chains predict mortality in patients with chronic kidney disease stage 5 on hemodialysis. METHODS: We performed a prospective cohort study of 160 hemodialysis patients with a median follow-up of 15 months (interquartile range, 3-44 months). Serum levels of free kappa and lambda immunoglobulin light chains were measured at the start of the study. The primary end point was mortality from any cause. RESULTS: In survivors, median serum levels of free kappa plus lambda immunoglobulin light chains were significantly higher compared with nonsurvivors (p < 0.05). Survival was significantly longer in those patients who had serum levels of free kappa plus lambda immunoglobulin light chains above the median compared with patients with serum levels below the median of 210 mg/l (chi(2) = 5.91; p = 0.015 by log-rank, Mantel-Cox, test). We performed univariate and multivariate regression analysis showing that older age and lower serum levels of free kappa plus lambda immunoglobulin light chains predicted mortality in hemodialysis patients. CONCLUSION: Higher serum levels of free kappa plus lambda immunoglobulin light chains ameliorate survival in hemodialysis patients. PMID- 21646815 TI - Fosinopril and losartan regulate klotho gene and nicotinamide adenine dinucleotide phosphate oxidase expression in kidneys of spontaneously hypertensive rats. AB - BACKGROUND/AIMS: Klotho, a newly identified antiaging gene, predominantly detected in the kidney, has pleiotropic protective effects on kidney diseases. Several studies have confirmed the association between Klotho and oxidative stress. The present studies were performed to explore effects of fosinopril (Fos) and losartan (Los) on Klotho and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase expression in kidneys of spontaneously hypertensive rats (SHR). METHODS: Twenty-four male 22-week-old SHR were randomly divided into three groups: model group, Fos group and Los group. Wistar-Kyoto rats were taken as control. After 8 weeks, urinary N-acetyl-beta-D-glucosaminidase (NAGase), 24 h urinary protein (Upro), serum creatinine (Scr), blood urea nitrogen (BUN) and renal pathological changes were detected. Renal mRNA and protein expression of Klotho and three subunits of NADPH oxidase protein expression were evaluated. RESULTS: As compared to the model group, NAGase, Upro, Scr and BUN were decreased; the typical renal pathological damage was relieved in the Fos or Los group. Fos or Los inhibited the reduction of Klotho expression, and reduced the elevation of NADPH oxidase expression. CONCLUSION: Abnormal expression of Klotho and NADPH oxidase plays important roles in progression of hypertensive renal damage. Fos and Los can increase Klotho expression, and inhibit NADPH oxidase expression, which may be one of the mechanisms of their protective effects in hypertensive renal damage. PMID- 21646816 TI - Long-term antihypertensive effect of angiotensin-converting enzyme inhibitory peptide LAP. AB - OBJECTIVE: To study the antihypertensive effects of the angiotensin-converting enzyme (ACE)-inhibitory peptide LAP on blood pressure in spontaneously hypertensive rats (SHRs). METHODS: A cohort of 12-week-old SHRs was randomly divided into 2 distinct groups, and ACE-inhibitory peptide LAP (experimental group) or physiological saline (controls) were administered. Caudal arterial blood pressure was then measured at specific time points (0, 4, 8 and 12 weeks). RESULTS: Systolic blood pressure of the SHRs showed a significant decrease after intraperitoneal injection with the ACE-inhibitory peptide LAP. Moreover, this depressurization effect lasted for over 1 month. CONCLUSION: Systolic blood pressure of SHRs could effectively be depressed in the long term by antihypertensive activity mediated by the ACE-inhibitory peptide LAP. PMID- 21646817 TI - Correlation of clinical changes with regard to thyroxine replacement therapy in hypothyroid patients: focusing on the change of renal function. AB - Thyroid dysfunction has an important role in renal insufficiency. The aim of the study was to correlate the change of renal function with other clinical factors after thyroxine therapy in hypothyroid patients. A prospective study was designed and 30 hypothyroid patients were included. All study subjects received 0.15-0.2 mg/day thyroxine for 12 weeks. Diastolic blood pressure and serum levels of creatine phosphokinase (CPK) and myoglobulin decreased significantly after thyroxine therapy. Serum creatinine decreased (0.87 +/- 0.22 vs. 0.70 +/- 0.17 mg/dl, p < 0.001) and estimated glomerular filtration rate (eGFR) increased significantly (82.06 +/- 31.08 vs. 100.31 +/- 31.79 ml/min/1.73 m(2); p < 0.001) after thyroxine replacement. Left ventricular ejection fraction (LVEF) was significantly increased after thyroxine replacement (64.47 +/- 11.94 vs. 72.40 +/ 13.89%, p = 0.026). No significant vascular functional changes of peripheral (pulse wave velocity) and renal interlobar arteries (pulsatility index and resistance index) were noted. The change of eGFR significantly correlated with the changes of serum-free T(4) (fT(4)), CPK, myoglobulin and LVEF. The correlation between the change of eGFR and thyroid-stimulating hormone (TSH) level was not significant. In conclusion, the GFR of hypothyroid patients increased significantly after thyroxine replacement. The change of GFR was significantly correlated with the changes of fT(4), CPK, myoglobulin and LVEF, but not with TSH. PMID- 21646818 TI - 4-Chloro-7-trifluoromethyl-10H- benzo[4,5]furo[3,2-b]indole-1-carboxylic acid (TBIC), a putative BK(Ca) channel opener with uterine relaxant activities. AB - In the present study, we examined the uterine relaxant activity of 4-chloro-7 trifluoromethyl-10H-benzo[4,5]furo[3,2-b]indole-1-carboxylic acid (TBIC), a putative opener of the large conductance Ca(2+)-activated K(+) (BK(Ca)) channel. TBIC concentration-dependently inhibited spontaneous uterine contractions (EC(50) = 4.63 MUmol/l; E(max) = 94.85 +/- 1.85%; 100 MUmol/l, n = 6). It also reduced contractions induced by oxytocin (EC(50) = 4.10 MUmol/l; E(max) = 84.3 +/- 3.83%; 100 MUmol/l, n = 6), prostaglandin F(2)(alpha) (EC(50) = 2.14 MUmol/l; E(max) = 73.70 +/- 5.21%; 100 MUmol/l, n = 6) and acetylcholine (EC(50) = 4.37 MUmol/l; E(max) = 83.67 +/- 4.82; 100 MUmol/l, n = 6). TBIC decreased KCl (20 mmol/l) induced contractions (EC(50) = 3.04 MUmol/l; E(max) = 94.0 +/- 3.12%; 100 MUmol/l, n = 6) indicating its K(+) channel opening activity. BK(Ca) channel blockers penitrem A (100 nmol/l) and tetraethylammonium chloride (1 mmol/l) attenuated the inhibitory activities of TBIC (p < 0.001) but not other K(+) channel blockers such as barium chloride and glibenclamide (K(IR) and K(ATP) channel blockers, respectively). These results demonstrate the uterine relaxant effects of TBIC in a mechanism of action largely referable to the potentiation of the BK(Ca) channels. We have provided evidence for the potential use of TBIC as a tocolytic agent and support for the utility of BK(Ca) channel openers in pathophysiologic conditions involving smooth muscle hyperactivity. PMID- 21646819 TI - Heme oxygenase-1 overexpression fails to attenuate hypertension when the nitric oxide synthase system is not fully operative. AB - Heme oxygenase (HO) is an enzyme that is involved in numerous secondary actions. One of its products, CO, seems to have an important but unclear role in blood pressure regulation. CO exhibits a vasodilator action through the activation of soluble guanylate cyclase and the subsequent production of cyclic guanosine monophosphate (cGMP). The aim of the present study was to determine whether pathological and pharmacological HO-1 overexpression has any regulatory role on blood pressure in a renovascular model of hypertension. We examined the effect of zinc protoporyphyrin IX (ZnPP-IX) administration, an inhibitor of HO activity, on mean arterial pressure (MAP) and heart rate in sham-operated and aorta-coarcted (AC) rats and its interaction with the nitric oxide synthase (NOS) pathway. Inhibition of HO increased MAP in normotensive rats with and without hemin pretreatment but not in hypertensive rats. Pretreatment with NG-nitro-L-arginine methyl ester blocked the pressor response to ZnPP-IX, suggesting a key role of NOS in the cardiovascular action of HO inhibition. In the same way, AC rats, an experimental model of hypertension with impaired function and low expression of endothelial NOS (eNOS), did not show any cardiovascular response to inhibition or induction of HO. This finding suggests that eNOS was necessary for modulating the CO response in the hypertensive group. In conclusion, the present study suggests that HO regulates blood pressure through CO only when the NOS pathway is fully operative. In addition, chronic HO induction fails to attenuate the hypertensive stage induced by coarctation as a consequence of the impairment of the NOS pathway. PMID- 21646820 TI - Effects of beraprost sodium, a prostaglandin I(2) analog, on high glucose-induced proliferation and oxidative stress in a rat glomerular mesangial cell line. AB - To investigate the effects of beraprost sodium on the proliferation and oxidative stress of glomerular mesangial cells under high glucose conditions, a rat mesangial cell line (rat mesangial cells; RMCs) was treated with beraprost sodium in the presence of high glucose concentrations. Proliferation rates of mesangial cells were detected by MTT assays and BrdU incorporation analyses. Levels of reactive oxygen species (ROS) were detected by DCFH-DA probes. The mRNA expression levels of CuZnSOD, MnSOD, catalase (CAT), glutathione peroxidase (Gpx), and collagen IV were detected by RT-PCR, and the protein levels of antioxidants (i.e. CuZnSOD, CAT, and MnSOD) and collagen IV were detected by Western blot. Beraprost sodium treatment significantly decreased the proliferation and ROS levels of RMCs cultured in high glucose conditions in a dose-dependent manner (p < 0.05). Beraprost sodium treatment decreased the mRNA and protein levels of CuZnSOD, CAT, and collagen IV in cells under high glucose conditions, while it increased MnSOD protein levels in cells under normal glucose conditions. Therefore, beraprost sodium inhibits high glucose-induced cellular proliferation and the generation of ROS, and it improves the antioxidant capacities of rat glomerular mesangial cells. PMID- 21646821 TI - Unipolar depression and the progression of coronary artery disease: toward an integrative model. AB - BACKGROUND: Despite extensive research on the relationship between depression and coronary artery disease (CAD) after an acute coronary syndrome (ACS), causal interpretations are still difficult. This uncertainty has led to much confusion regarding screening and treatment for depression in CAD patients. METHOD: A critical and conceptual analysis of the pertinent literature, which elaborates the implications of the heterogeneity in symptom pattern, etiology, and course of depression in CAD patients. RESULTS: We propose an integrative dynamic model of the depression-CAD relationship. The model rests on three core hypotheses: (1) Depression in CAD patients consists of mixtures of two types of depression, denoted as 'cognitive/affective' and 'somatic' depression, each having a somewhat characteristic symptom expression and etiology. (2) Effects of depression on CAD depend on the type and duration of depression. The dynamic aspect of the model indicates that post-ACS depression shifts, when it persists, from a marker of the severity (somatic type) and meaning (cognitive/affective type) of the ACS to a largely indirect causal factor in the progression of CAD. (3) The most plausible pathways mediating the effects of persistent/recurrent depression, irrespective of type, on cardiac prognosis are behavioral and act by making depressed CAD patients more susceptible to other CAD risks. The model offers testable predictions and explanations for a variety of apparently unrelated or inconsistent findings. CONCLUSION: The proposed model may have potential for integrating findings regarding the depression-CAD relationship, contributing to the clarification of discords on screening and treatment of depression, and guiding future research. PMID- 21646822 TI - Quality of life in medically ill persons with comorbid mental disorders: a systematic review and meta-analysis. AB - BACKGROUND: This systematic review aims to investigate the association between comorbid mental disorders and quality of life (QoL) in patients with chronic medical diseases. METHODS: Studies investigating adults with diabetes mellitus, coronary artery disease, asthma, chronic back pain and colorectal cancer were included. Two reviewers independently extracted data and assessed methodological criteria. Effect sizes for QoL scores were analyzed in random-effects meta analyses. Subgroup and sensitivity analyses were conducted. RESULTS: The database search identified 7,291 references and 65 primary studies were included. Medically ill persons with comorbid mental disorders showed a significantly decreased overall (d = -1.10; 95% CI = -1.34 to -0.86), physical (d = -0.64; 95% CI = -0.74 to -0.53) and psychosocial (d = -1.18; 95% CI = -1.42 to -0.95) QoL compared to persons without mental disorders. Subgroup analyses did not reveal significant differences between the examined medical diseases or mental disorders. CONCLUSION: The review provides evidence of a substantially reduced psychosocial and physical QoL in medically ill patients with comorbid mental disorders. This patient-reported outcome highlights the importance of recognizing and treating comorbid mental disorders in the medically ill. PMID- 21646823 TI - Motivational intervention to reduce cannabis use in young people with psychosis: a randomized controlled trial. AB - BACKGROUND: Cannabis use has a negative impact on psychosis. Studies are needed to explore the efficacy of psychological interventions to reduce cannabis use in psychosis. Our aim is to study the efficacy of a specific motivational intervention on young cannabis users suffering from psychosis. METHODS: Participants (aged less than 35 years) were randomly assigned to treatment as usual (TAU) alone, or treatment as usual plus motivational intervention (MI + TAU). TAU was comprehensive and included case management, early intervention and mobile team when needed. Assessments were completed at baseline and at 3, 6 and 12 months follow-up. RESULTS: Sixty-two participants (32 TAU and 30 MI + TAU) were included in the study. Cannabis use decreased in both groups at follow-up. Participants who received MI in addition to TAU displayed both a greater reduction in number of joints smoked per week and greater confidence to change cannabis use at 3 and 6 months follow-up, but differences between groups were nonsignificant at 12 months. CONCLUSIONS: MI is well accepted by patients suffering from psychosis and has a short-term impact on cannabis use when added to standard care. However, the differential effect was not maintained at 1-year follow-up. MI appears to be a useful active component to reduce cannabis use which should be integrated in routine clinical practice. PMID- 21646824 TI - Computerized testing of neurocognitive function in euthymic bipolar patients compared to those with mild cognitive impairment and cognitively healthy controls. AB - OBJECTIVES: While neuropsychological impairment in bipolar disorder is well documented, the effect size of this impairment is rarely compared directly to that in other clinically familiar cognitive disorders. This study compares neuropsychological functioning of euthymic bipolar patients to those with mild cognitive impairment (MCI) as well as healthy controls. METHODS: Following evaluation during regular follow-up in a mood disorders clinic, 58 euthymic adult bipolar subjects were administered a validated and fully computerized cognitive assessment (Mindstreams; NeuroTrax Corp., N.Y., USA). Study data were compared to existing data for MCI and cognitively healthy individuals tested with the same assessment. RESULTS: Final analyses were based on 51 bipolar patients, 162 MCI patients and 495 healthy comparison subjects. Significant (p < 0.001) group effects were found for every parameter. Post hoc analysis revealed that the bipolar and MCI groups showed statistically equivalent functioning in memory, executive function, verbal function, and information processing speed. In the domains of visual-spatial processing, attention, and motor skills, the MCI group outperformed the bipolar group. In every domain, the healthy control group outperformed both the bipolar and the MCI groups. CONCLUSIONS: The cognitive function of euthymic bipolar patients and those diagnosed with MCI was found to be similar in most but not all domains. Both groups performed significantly less well than the comparison group of healthy subjects. It may be helpful for clinicians to conceptualize the overall level of cognitive impairment in bipolar patients as similar to that in MCI. PMID- 21646825 TI - Uptake of antioxidants by natural nutrition and supplementation: pros and cons from the dermatological point of view. AB - The pros and cons of the systemic and topical application of antioxidant substances are a subject of intense discussion among experts, with resulting confusion for consumers and producers. The objective of the present article is to clarify the various uncertainties relating to the use of antioxidant substances in dermatology. Whereas inappropriate application of antioxidant substances (concerning their concentration and composition) might induce harmful effects, the consumer will definitively benefit from physiological concentrations and compositions of antioxidants. The most suitable method is the consumption of natural antioxidants in the form of fruit and vegetables, for example. In addition, the skin, which also accumulates antioxidant substances, may profit from a sufficient antioxidative level, as damage induced by sun radiation in addition to skin aging is reduced. PMID- 21646826 TI - [Regional distribution of all-cause mortality and coronary disease incidence in Turkey: findings of Turkish Adult Risk Factor survey 2010]. AB - OBJECTIVES: We analyzed the distribution of cumulative all-cause and cardiovascular mortality and incident coronary heart disease (CHD) across the seven geographic regions of Turkey and presented overall and coronary mortality findings of the 2010 survey of the Turkish Adult Risk Factor Study. STUDY DESIGN: A total of 1406 participants were surveyed. Information on the mode of death was obtained from first-degree relatives and/or health personnel of local heath offices. Information on survivors was obtained from history, physical examination, and 12-lead electrocardiography. RESULTS: Of the surveyed participants, 686 were examined; information on health status was obtained in 577 subjects, and 32 participants (14 women, 18 men; mean age 72.3+/-15.6 years) were ascertained to have died. The total duration of follow-up was 2,520 person-years. Nineteen deaths were of coronary (n=16) or cerebrovascular (n=3) origin. Cumulative 20-year assessment of the entire cohort for the age bracket of 45-74 years disclosed a high coronary mortality rate, being 7.4 and 4.1 per 1000 person years in men and women, respectively, and representing a limited decline after year 2000. Age-adjusted Cox regression analysis comprising 433 deaths and 506 incident CHD cases over a 7.3-year follow-up showed similar mortality rates across the regions, and a significantly high CHD incidence in males of the Black Sea and Marmara regions and in females of the Southeast Anatolia. Currently, 480,000 incident CHD cases are estimated yearly in Turkey. CONCLUSION: The high age-adjusted overall mortality in Turkey shows nonsignificant differences across geographic regions, whereas the age-adjusted CHD incidence is high in the Black Sea and Marmara regions. PMID- 21646827 TI - [The relation of serum monocyte chemoattractant protein-1 level with coronary atherosclerotic burden and collateral degree in stable coronary artery disease]. AB - OBJECTIVES: We investigated whether serum monocyte chemoattractant protein-1 (MCP 1) level predicted coronary atherosclerotic burden in patients with stable coronary artery disease and its relationship with coronary collateral grade. STUDY DESIGN: We prospectively included 196 patients (103 males, 93 females; mean age 59 +/- 11 years) who underwent coronary angiography for stable angina pectoris. Serum MCP-1 levels were determined before coronary angiography. Coronary atherosclerotic burden was measured by the Gensini score, and coronary collateral development was assessed by the Rentrop classification. The patients were divided into four groups: those with normal coronary arteries (NCA); those with coronary lesions of <70% luminal obstruction; and those with coronary lesions of >= 70% luminal obstruction accompanied by a good or poor collateral grade. RESULTS: The mean serum MCP-1 level was higher in patients with coronary lesions compared to patients with NCA (129 +/- 130 vs. 102 +/- 55 pg/ml, p=0.048). Although there were no significant differences in the MCP-1 levels of patients with NCA, with <70% luminal obstruction, and those with a significant luminal obstruction and a poor collateral grade, patients with significant luminal obstruction and a good collateral grade had significantly higher MCP-1 levels compared to the remaining groups (p=0.016). However, in multivariate regression analysis, MCP-1 level was not independently associated with the Gensini score. CONCLUSION: Our findings suggest that serum MCP-1 level is higher in patients with coronary atherosclerosis, without a significant and independent association with coronary atherosclerotic burden. Significantly increased serum MCP-1 levels in patients with a good collateral grade may be an interesting issue of investigation. PMID- 21646828 TI - [The impact of chronic kidney disease on in-hospital clinical outcomes in patients undergoing primary percutaneous angioplasty for ST-segment elevation myocardial infarction]. AB - OBJECTIVES: We investigated the effect of chronic kidney disease (CKD) on in hospital results in patients undergoing primary percutaneous angioplasty for ST segment elevation myocardial infarction (STEMI). STUDY DESIGN: The study included 2,486 patients (2,070 men, 416 women) who were treated with primary angioplasty for STEMI. Of these, 273 patients (11%) were found to have CKD (glomerular filtration rate <60 ml/min/1.73 m2) before the procedure. Patients with and without CKD were evaluated with respect to demographic and clinical features, primary angioplasty findings, and in-hospital clinical results. RESULTS: Patients with CKD exhibited a higher mean age, Killip class, and higher frequencies of female gender, diabetes, hypertension, anemia, and previous myocardial infarction (p<0.05). Angioplasty showed higher rates of right coronary artery lesion, multivessel disease, contrast nephropathy, unsuccessful procedure, and increased stenosis rate and stent length in CKD patients (p<0.05). Cardiovascular mortality occurred in 11.7% and 1.4% of patients with and without CKD, respectively (p<0.001). Patients with CKD had significantly higher incidences of target vessel revascularization, major cardiac events, stroke, cardiopulmonary resuscitation, hemodialysis, ventricular tachycardia/fibrillation, severe heart failure, cardiogenic shock, and significant hemorrhage (p<0.05). Multivariate analysis showed that CKD was an independent predictor of mortality (OR=4.1, 95% CI 1.83 9.17; p=0.001). CONCLUSION: Our findings show that CKD patients undergoing primary angioplasty for STEMI have an increased risk profile and poorer in hospital results, and that CKD represents an independent risk factor for mortality. PMID- 21646829 TI - [The incidence of thrombocytopenia after cardiopulmonary bypass in children with Down syndrome]. AB - OBJECTIVES: We investigated the incidence of thrombocytopenia in pediatric patients with Down syndrome following cardiac surgery for congenital heart disease. STUDY DESIGN: We retrospectively evaluated 162 patients (81 girls, 81 boys; mean age 26.1 +/- 39.5 months) who underwent total surgical correction for congenital heart disease. The patients were divided into two groups with respect to the presence (n=118) or absence (n=44, controls) of Down syndrome. Platelet counts were performed preoperatively and on days 1 to 7 after surgery and thrombocytopenia was defined as a platelet count of less than 100,000/mm3. RESULTS: The incidence of thrombocytopenia was significantly higher in patients with Down syndrome compared to controls (61.9% vs. 34.1%, p=0.002). Severe thrombocytopenia was observed in 22% and 4.6% of cases with and without Down syndrome, respectively. Postoperative platelet counts showed sharp decreases in both groups, bottoming out on day 3 and with more significant decreases in patients with Down syndrome; they started to rise on day 4, but remained lower than baseline levels on day 7. The only significant differences between the two groups in preoperative and postoperative variables were higher incidences of reintubation (26.3% vs. 9.1%, p=0.003) and complications during intensive care (50.9% vs. 27.3%, p=0.007) in cases with Down syndrome. Comparison of patients with and without thrombocytopenia independent of Down syndrome yielded significant differences for thrombocytopenic patients with respect to age, operation age, the presence of cyanosis, type of surgery and operation time, aortic clamp and bypass times, lengths of intensive care and hospital stay, intubation and chest tube drainage times, and the incidence of postoperative complications (p<0.05). CONCLUSION: Despite higher incidence of thrombocytopenia, the presence of Down syndrome was not associated with significant differences other than increased reintubation requirement and higher complication rate during intensive care. PMID- 21646830 TI - [Cardiac autonomic function and cardiac arrhythmias in patients with obstructive sleep apnea]. AB - OBJECTIVES: We assessed the effect of obstructive sleep apnea syndrome (OSAS) on cardiac autonomic function with the parameters of heart rate variability (HRV) and heart rate turbulence (HRT) and investigated the frequency of cardiac arrhythmias in patients with OSAS. STUDY DESIGN: Seventy-six patients (27 women, 49 men; mean age 50.6+/-10.7 years; range 33 to 75 years) with an initial diagnosis of OSAS underwent polysomnography and simultaneous Holter monitoring. The diagnosis of OSAS was based on an apnea-hypopnea index (AHI) of >=5. The patients were evaluated with time-and frequency-based parameters of HRV, and turbulence onset (TO) and turbulence slope (TS) parameters of HRT. Cardiac arrhythmias were also assessed on Holter monitoring. RESULTS: Fifty-three patients (69.7%) were diagnosed with OSAS, being mild in 25 (47.2%), moderate in 23 (43.4%), and severe in five (9.4%) patients based on the AHI. The patients were divided into two groups as those having an AHI of <5 (n=23) and >=5 (n=53). There were no significant differences in HRV parameters between the two groups, but patients with OSAS exhibited a significantly higher TO (p=0.02) and a significantly lower TS (p<0.001). In bivariate correlation analysis, AHI showed a significant negative correlation with TS (r=-0.37, p=0.009) and a significant positive correlation with TO (r=0.36, p=0.01). The frequencies of short-sequence premature atrial and ventricular contractions were significantly higher in patients with OSAS (p<0.001). CONCLUSION: Our findings suggest that blunting of HRT is more prominent than changes in HRV parameters in patients with OSAS, accompanied by increased frequency of cardiac arrhythmias. PMID- 21646831 TI - Primary angioplasty in a high-volume tertiary center in Turkey: in-hospital clinical outcomes of 1625 patients. AB - OBJECTIVES: We evaluated in-hospital results of primary percutaneous coronary intervention (PCI) in a high-volume tertiary center. STUDY DESIGN: We retrospectively evaluated 1625 patients (1323 males, 302 females; mean age 56.0 +/- 11.6 years) who underwent primary PCI for acute ST-elevation myocardial infarction between January 2006 and April 2008. All coronary angiography procedures were performed using the femoral artery route. In-hospital clinical and angiographic results were recorded. RESULTS: On admission, 23% of the patients had diabetes mellitus, 49.6% had anterior myocardial infarction, and 4.9% had cardiogenic shock. The mean duration of pain was 171.2 +/- 121.2 minutes, and the mean door-to-balloon time was 31.6 +/- 7.2 minutes. Infarct related artery was the left anterior descending artery in 49.7%, multivessel disease was present in 40.9%, TIMI 2/3 flow was present in 23.6%, and high-grade thrombus was observed in 66.8%. Primary PCI involved balloon dilatation (5.7%) and stent implantation (94.3%). The incidence of angiographic no-reflow was 11.9%. The mean hospital stay was 5.2 +/- 3.3 days. All-cause mortality occurred in 71 patients (4.4%). Other in-hospital events were reinfarction (1.4%), target vessel revascularization (1.9%), hemorrhagic/ischemic stroke (0.6%), stent thrombosis (1.2%), major bleeding (3.8%), blood transfusion (4.8%), heart failure (10.5%), atrial fibrillation (4%), and ventricular tachycardia (3.9%). CONCLUSION: Primary PCI is an effective method in achieving complete revascularization of the infarct-related artery. Successful in-hospital results not only depend on the experience and equipment of the center, but also on how rapidly reperfusion is achieved. PMID- 21646832 TI - ST-elevation myocardial infarction due to a spontaneous thrombus in the left anterior descending artery in a young HIV-infected patient. AB - With increasing life expectancy due to highly active antiretroviral therapy (HAART), the spectrum of human immunodeficiency virus (HIV)-associated morbidity and mortality has shifted from opportunistic infections toward associated chronic medical conditions. We report on a 26-year-old female patient receiving HAART for HIV infection, who developed spontaneous thrombosis of the proximal left anterior descending (LAD) artery, resulting in acute ST-elevation myocardial infarction. She had none of the conventional risk factors for the development of coronary artery disease. Following diagnostic coronary angiography that showed a large (16x3.4 mm) spontaneous thrombus in the proximal LAD artery, percutaneous coronary intervention was performed with prior aspiration of the occluding thrombus and implantation of a bare-metal stent. The patient was discharged with instruction of appropriate medical therapy. This case highlights the association between immunosuppression with HAART, particularly protease inhibitors, and the development of accelerated atherosclerosis in patients with HIV infection. PMID- 21646833 TI - A very rare combination of four coronary artery anomalies in a patient with acute inferior myocardial infarction. AB - We present a combination of four rarely seen coronary anomalies: double right coronary artery originating from the right coronary sinus (RCS) and left main coronary artery, respectively, and separate origination of the left anterior descending (LAD) artery, circumflex artery (Cx), and septal perforator artery from the RCS. These anomalies were encountered in a 46-year-old male patient who had a previous diagnosis of spina bifida occulta and renal pelvis and presented with the complaint of chest pain of two-hour onset. He had no conventional coronary risk factors and no history of chest pain or syncope. Electrocardiography showed ST-segment elevation and cardiac enzyme levels were elevated. Coronary angiography was performed with the diagnosis of acute inferior myocardial infarction, which showed a severe stenosis in the mid portion of the LAD and total occlusion in the proximal part of the Cx. Balloon dilatation and stent implantation were performed for the Cx lesion and TIMI 3 flow was achieved. One month after the procedure, percutaneous coronary intervention was repeated for the LAD lesion and patency was achieved with balloon dilatation and stenting. Since visualization of all the coronary anomalies mentioned above posed some difficulties during coronary angiography, cardiac computed tomography angiography was also used to reveal the ostia and the courses of coronary arteries. This combination of four rare coronary anomalies has not been reported before. PMID- 21646834 TI - An unusual microorganism, Aerococcus viridans, causing endocarditis and aortic valvular obstruction due to a huge vegetation. AB - Aerococcus viridans is not a common pathogen, and endocarditis due to A. viridans is very rare. A 44-year-old woman with persistent atrial fibrillation and rheumatic valvular heart disease was admitted with fever, sweating, weakness, and progressive shortness of breath. Transthoracic echocardiography (TTE) demonstrated a 8x9-mm vegetation attached to the right coronary cusp of the aortic valve, causing aortic obstruction. Blood cultures yielded A. viridans susceptible to penicillin. Despite optimal antibiotherapy, subsequent TTE controls revealed enlargement of the vegetation, reaching a size of 21x10 mm, and an increasing gradient across the aortic valve. The patient underwent successful aortic and mitral valve replacement and was stable in the postoperative period without any problem. This represents the first reported case of A. viridans endocarditis in which the size and location of vegetation caused obstruction to blood flow, indicating surgery. PMID- 21646835 TI - Right atrial myxoma associated with portal and splenic vein thrombosis in a patient with Budd-Chiari syndrome. AB - Budd-Chiari syndrome (BCS) is a rare disorder characterized by hepatic venous obstruction. A 41-year-old male patient presented with right upper quadrant pain, abdominal distension, and dyspnea. He had a history of BCS that was associated with polycythemia vera. Abdominal computed tomography showed hepatomegaly and a hypodense filling defect suggestive of thrombus formation in the hepatic, splenic, and portal veins, and suprahepatic part of the inferior vena cava. Transthoracic echocardiography performed to assess the extension of this pathological process and its relation with intracardiac structures showed a mass lesion in the right atrium, about 4 x 3 cm in diameter. The lesion manifested as an intracardiac thrombus extending from the inferior vena cava. The patient underwent surgical treatment to remove the atrial mass. At surgery, the lesion turned out to be an atrial tumor, which was diagnosed as atrial myxoma in histopathologic examination. The symptoms of the patient resolved after surgery. PMID- 21646836 TI - [Right coronary artery arising from the distal left circumflex artery]. AB - Origination of the right coronary artery from the distal left circumflex artery is a rare anomaly. A 63-year-old woman was admitted with subacute anteroseptal myocardial infarction. Electrocardiography showed a QS pattern in V1-V3 precordial leads without ST elevation. Cardiac enzyme levels were elevated (CK-MB 186 ng/ml, troponin I 27.1 ng/ml). Echocardiography showed hypokinesia of the anterior and lateral walls without valvular pathology. Coronary angiography revealed origination of the right coronary artery from the circumflex artery. The right coronary artery had a normal flow pattern and there were atherosclerotic plaques in the circumflex artery without a significant stenosis. Distal to the first diagonal branch of the left anterior descending artery, a 95% stenotic lesion was detected, which was treated with balloon dilatation followed by implantation of a bare metal stent. The patient was discharged with near-complete patency and without any complication. PMID- 21646837 TI - [Dilated cardiomyopathy associated with dystrophic epidermolysis bullosa: role of micronutrient deficiency?]. AB - Autosomal recessive dystrophic epidermolysis bullosa (DEB) is a chronic skin disorder characterized by widespread bullous formation, erosions, and scar formation. There have been reports of dilated cardiomyopathy and death in patients with DEB. The pathogenesis of cardiomyopathy in DEB remains uncertain, but some drugs, viral infections, iron loading, micronutrient deficiencies such as selenium and carnitine have been implicated. A 16-year-old boy who was followed-up from birth with the diagnosis of DEB presented with respiratory distress and heart failure symptoms of two-week history and early fatigue within the past year. Etiological evaluation showed a low plasma selenium level. Echocardiographic examination yielded the diagnosis of dilated cardiomyopathy. Findings of viral serology tests and metabolic screening were normal. Selenium replacement and anticongestive treatment were initiated, which led to partial improvement in cardiac functions. The authors draw attention to the possible role of micronutrient deficiency in the development of cardiomyopathy in patients with DEB. PMID- 21646838 TI - Transradial and transulnar access for percutaneous coronary interventions. AB - Periprocedural bleeding and vascular complications after percutaneous coronary intervention (PCI) are associated with worse clinical outcomes and increased short- and long-term mortality. Vascular access-related bleeding accounts for more than 80% of all major bleeding events in PCI performed by the transfemoral approach. Transradial approach (TRA), on the other hand, virtually eliminates access site bleeding and vascular complications. Although clinical trials have mostly evaluated different pharmacological strategies for reducing bleeding risk, adoption of a radial rather than a femoral access may allow greater reductions in bleeding complications than pharmacological strategies alone. High-risk patients such as those with acute coronary syndrome and ST-segment elevation myocardial infarction, women, obese patients, and elderly subjects who are at increased risk for vascular complications and bleeding might particularly benefit from the radial approach. Besides increased patient safety, the TRA is associated with improved patient satisfaction, reduced cost, and length of hospital stay, thus allowing outpatient performance of uncomplicated PCI. PMID- 21646839 TI - Congenital left ventricular diverticulum. PMID- 21646840 TI - Angiographic catheter-induced subclavian artery spasm. PMID- 21646841 TI - Surgical treatment of an anomalous left coronary artery from the pulmonary artery. PMID- 21646842 TI - Unidirectional intercoronary communication: a very rare coronary anomaly and cause of ischemia. PMID- 21646843 TI - [Calcific constrictive pericarditis]. PMID- 21646844 TI - A case of myocarditis mimicking acute coronary syndrome associated with H1N1 influenza A virus infection. PMID- 21646845 TI - A case of twin circumflex arteries associated with acute myocardial infarction. PMID- 21646846 TI - Discovery of the serum biomarker proteins in severe preeclampsia by proteomic analysis. AB - Preeclapsia (PE) is a severe disorder that occurs during pregnancy, leading to maternal and fetal morbidity and mortality. PE affects about 3-8% of all pregnancies. In this study, we conducted liquid chromatography- mass spectrometry/mass spectrometry (LC-MS/MS) to analyze serum samples depleted of the six most abundant proteins from normal and PE-affected pregnancies to profile serum proteins. A total of 237 proteins were confidently identified with <1% false discovery rate from the two groups of duplicate analysis. The expression levels of those identified proteins were compared semiquantitatively by spectral counting. To further validate the candidate proteins with a quantitative mass spectrometric method, selective reaction monitoring (SRM) and enzyme linked immune assay (ELISA) of serum samples collected from pregnant women with severe PE (n = 8) or normal pregnant women (n = 5) was conducted. alpha2- HS glycoprotein (AHSG), retinol binding protein 4 (RBP4) and alpha-1 microglobulin/bikunin (AMBP) and Insulin like growth factor binding protein, acid labile subunit (IGFBP-ALS) were confirmed to be differentially expressed in PE using SRM (P<0.05). Among these proteins, AHSG was verified by ELISA and showed a statistically significant increase in PE samples when compared to controls. PMID- 21646847 TI - [Changes in serum prostate specific antigen and testosterone levels after chlormadinone acetate treatment in patients with benign prostatic hyperplasia : a prospective multicenter clinical study]. AB - In this prospective multicenter study, we investigated the changes in serum prostate-specific antigen (PSA) and testosterone levels after treatment with antiandrogen chlormadinone acetate (CMA) in patients with benign prostatic hyperplasia (BPH). The inclusion criteria for the patients were as follows : PSA value of C10 ng/ml, maximum urine flow rate of <15 ml/s, estimated prostate volume of B20 ml, International Prostate System Score (IPSS) of B8, and IPSS quality of life (QOL) index of B2. Of the 115 patients who registered, 114 qualified for this study. The patients were treated with CMA (50 mg/day) for 16 weeks ; this was followed by a no-CMA phase of 32 weeks. When compared with the baseline PSA level, the levels at 8 and 16 weeks of treatment had decreased by 56.4% (95% confidence interval [CI], 51.1-1.2) and 57.6% (95% CI, 52.3-62.4), respectively. Similarly, when compared with the baseline testosterone level, the levels at 8 and 16 weeks of treatment had decreased by 90.1% (95% CI, 87.8-91.9) and 84.4% (95% CI, 80.7-87.4), respectively. After treatment discontinuation, the PSA levels gradually increased and returned to baseline in 32 weeks. However, the testosterone levels returned to baseline in only 8 weeks. Although patients over 80 years of age showed a gradual decrease in these levels when compared with younger patients, the changes in the levels of PSA and testosterone were not affected by age. Thus, in order to use antiandrogen agents including CMA for treating BPH, we need to determine the PSA value that converted it into double. PMID- 21646848 TI - [A case of amyloidosis of the ureter caused by primary macroglobulinemia]. AB - A 78-year-old woman was referred to our hospital with chief complaint of colicky right flank pain. Computed tomography (CT) demonstrated right hydronephrosis and a wall thickening in the right ureteropelvic junction. Right nephroureterectomy was performed under a clinical diagnosis of invasive ureteral cancer. Histological examination revealed immunoglobulin light chain (AL) amyloidosis of the ureter. Although amyloid deposition seemed to be localized to the ureter alone, bone marrow biopsy revealed primary macroglobulinemia. After 23 months, no signs or symptoms suggesting the progression of amyloidosis or macroglobulinemia have been pointed out. AL amyloidosis arising secondarily to plasma cell disorder is called immunocytic amyloidosis. In general, immunocytic amyloidosis accompanies deposition of the amyloid protein to various organs of the whole body, and causes multiple clinical symptoms related to amyloid deposition. Even though the clinical manifestation indicates the localized disease like the presented case, close examination of immunocytic amyloidosis is important. PMID- 21646849 TI - [Successful treatment of a case of advanced adult Wilms' tumor with pre-surgical chemotherapy, irradiation and radical nephrectomy]. AB - A 37-year-old male was referred with a huge renal mass and multiple lung lesions. Abdominal and chest computed tomographic scan revealed a 11*15*17. 5 cm right renal tumor and multiple lung metastases. Neither tumor was enhanced, and no lymph adenopathy was detected. Percutaneous needle core biopsy was performed and the tumors were diagnosed as Wilms' tumor histologically supported by immunohistological positive staining to WT-1 and CD56. Since lung metastases were detected and the renal tumor reached the abdominal aorta on the left side, presurgical systemic chemotherapy (ifosphamide+ carboplatin+etoposide ; ICEx 4 courses) was performed. The renal tumor and the lung metastases were markedly decreased in size and subjected to a radical nephrectomy. Histological examination confirmed the diagnosis of Wilms' tumor without anaplastic change categorized as mesenchymal type. As an adjuvant therapy, irradiation to renal bed (40 Gy/20 fr) and bilateral lung (12 Gy/8 fr) were performed. Six months after the end of the therapy, no evidence of recurrence was detected. PMID- 21646850 TI - [Pseudohypoaldosteronism type 1 in an infant with bilateral primary obstructive megaureter : a case report]. AB - A 35-day-old male infant was referred to our hospital for bilateral hydroureteronephrosis on postnatal ultrasonography. He was born at 37 weeks and 2 days of gestation, weighing 2,765 g. He was diagnosed with bilateral primary obstructive megaureter. During observation, at 3 months and 2 weeks of age, he presented with dehydration and showed hyponatremia and hyperkalemia. An endocrinological examination led to a diagnosis of pseudohypoaldosteronism type 1 (PHA1). After bilateral percutaneous nephrostomy, PHA1 resolved. He underwent bilateral ureteral tapering and ureteroneocystostomy at 5 months and 3 weeks of age. Since PHA1 recurred after removal of urethral catheter, urethral catheter and bilateral double-J-stent was kept until 8 months old. He was followed-up until 3 years old with no signs of PHA1 recurrence. For an infant with obstructive uropathy or vesicoureteral reflux showing hyponatremia and hyperkalemia, PHA1 should be considered. PMID- 21646851 TI - [A case of carcinosarcoma of the urinary bladder]. AB - A 66-year-old woman visited our hospital complaining of painful, irritative urinary symptoms and macroscopic hematuria. Cystoscopy revealed a non-papillary tumor covered with necrotic tissue on the right side of the posterior wall of the bladder. Transurethral resection was performed ; histologically, the tumor was found to be composed of carcinomatous and sarcomatous elements. The carcinomatous element consisted of urothelial and squamous cell carcinomas. The sarcomatous element was composed of osteosarcoma, chondrosarcoma and spindle cell sarcoma. Immunohistochemical examination showed that the carcinomatous component was positive for cytokeratin and the sarcomatous component was positive for S-100 protein. The patient underwent total cystectomy with ileal conduit under the diagnosis of carcinosarcoma. Pathological examination showed no residual tumor. She was followed up with no signs of recurrence or metastasis. Computed tomography (CT) at nine months following surgery showed no evidence of recurrence. However, thirteen months after the operation, she complained of lower abdominal pain, and CT demonstrated a bulky intrapelvic tumor and right hydronephrosis. Her condition worsened rapidly and she died one month later. PMID- 21646852 TI - [Long-term disease stabilization by docetaxel plus estramustine for castration resistant prostate cancer : report of a case]. AB - Chemotherapy with docetaxcel (DTX) plus estramustine (EMP) for castration resistant prostate cancer (CRPC) was started 30 months after the patient, a 65 year-old man, was diagnosed as having advanced prostate cancer cT3aN1M1 (OSS) with an initial PSA of 490 ng/ml. Prostate biopsy specimens revealed moderately differentiated adenocarcinoma, Gleason's sum 4+5. He was treated with DTX 30 mg/m2 on day 2 and oral EMP 560 mg/day days 1-3 weekly for 3 out of 4 weeks. PSA at start of DTX plus EMP was 81.7 ng/ml, and that after 59 months was 66.6 ng/ml. No objective change in computed tomography and bone scan were observed. He also had no cancer-related symptoms and activity of daily life was good. Chemotherapy was interrupted twice because of pleural effusion and dyspnea by DTX, at 3 and 4 months, respectively, long-term disease stabilization was obtained by this treatment. Other adverse events including interstitial pneumonia, cardiovascular disorders and myelosuppression were not observed. He was maintained on the same chemotherapy. DTX plus EMP chemotherapy is an effective treatment for CRPC patients. Continuing this therapy it is important to survey and control adverse events caused by DTX and EMP carefully. PMID- 21646853 TI - [Successful combined treatment with maximal androgen blockade (MAB), intra arterial chemotherapy and radiation for advanced prostatic ductal adenocarcinoma : report of two cases]. AB - The first case was in a 73-year-old man with macrohematuria. The second case was in a 59-year-old man with pollakiuria. Their serum prostate specific antigen levels were slightly elevated and urinary cytology was negative. Histological examination by prostatic needle biopsy and biopsy from bladder neck showed prostatic ductal adenocarcinoma. Clinical stage on computed tomography and magnetic resonance imaging was T4N0M0 in both cases. After 10-month maximal androgen blockade(MAB) and arterial chemotherapy using reservoir system, radiation therapy was performed. After that, low dose FP-chemotherapy(5 fluorouracil 600 mg/day, cisplatinum 10 mg/day) was performed for 28 days in the first case. At present, there are no signs of recurrence or metastasis in either case. PMID- 21646854 TI - [Renal autotransplantation for the treatment of nutcracker phenomenon which caused varicocele rupture : a case report]. AB - A 31-year-old man visited our hospital complaining of swelling in the left scrotum. Five days previously, he had felt sudden pain in the left lower abdomen and noticed swelling in the left scrotum. He had been suffering from intermittent gross hematuria and left flank pain for 1 year. An elastic hard mass was palpable in the left scrotum. Scrotal hematoma and marked dilation of left renal vein and left gonadal vein were revealed by computed tomography. A diagnosis of varicocele rupture secondary to nutcracker phenomenon was made. One month later, he underwent retroperitoneal laparoscopic donor nephrectomy and subsequent renal autotransplantation into the left iliac fossa. The post-operative course was uneventful. Gross hematuria was resolved 3 days after the operation, and swelling in the left scrotum was resolved within 6 months of the operation. Surgical intervention to treat nutcracker phenomenon is considered controversial. We believe that our procedure is a reliable option for surgical treatment for nutcracker phenomenon. PMID- 21646855 TI - Prognostic value of post-thyroidectomy thyroglobulin levels in patients with differentiated thyroid cancer. AB - BACKGROUND: Thyroglobulin is an excellent biological marker of persistent or recurrent thyroid cancer during long-term follow-up. Most studies investigated its diagnostic value but not its prognostic value over time. We aim to study the prognostic value of thyroglobulin levels early after total thyroidectomy, before iodine ablation. METHODS: The study was based on the Rabin Medical Center registry of patients with non-medullary thyroid carcinoma. Data were collected on the clinical, laboratory, and outcome characteristics of 420 consecutive patients followed at our institution for whom early post-operative pre-ablation thyroglobulin values (baseline thyroglobulin) were available. RESULTS: Patients were classified into 4 groups by baseline thyroglobulin level: 0-2, 2-10, 10-100, and >100 ng/ml. Higher levels were associated with a shift toward male gender (p=0.01), larger tumor size (p=0.02), and a more extensive disease (p<0.0001). They were also related to disease persistence and evidence of disease at last follow-up (p<0.0001). The 10 ng/ml cut-off level identified patients with persistent disease with a sensitivity and specificity of 73%, positive predictive value of 43%, and negative predictive value of 89%. On multivariate analysis, the following variables were predictive of persistent disease: baseline thyroglobulin level, male gender, lymph-node involvement, distant metastases, higher tumor invasiveness, and larger tumor size. However, the predictive power of baseline thyroglobulin level was relatively weak (odds ratio 1.002, 95% confidence interval 1.00-1.04). CONCLUSIONS: In patients with well-differentiated thyroid cancer, a post-thyroidectomy thyroglobulin level <10 ng/ml is associated with a low probability of having persistent disease and can be used combined with other disease characteristics for decisions regarding treatment and follow-up. PMID- 21646856 TI - Association among serum uric acid, cardiovascular risk, and arterial stiffness: a cross-sectional study in She ethnic minority group of Fujian Province in China. AB - OBJECTIVES: To investigate the association between serum uric acid levels, cardiovascular risk, and arterial stiffness in She ethnic minority in South China's Fujian Province. METHODS: 5109 participants aged 20-79 yr were enrolled in analysis. Tpeak-Tend interval (Tp-e), QT interval, and height of the R wave in lead aVL (aVLR) were measured on 12-lead electrocardiogram (ECG), and Minnesota code-indicated major abnormal ECG MA-ECG was used as a risk indicator of cardiovascular disease. Arterial stiffness was assessed by brachial-ankle pulse wave velocity (baPWV). RESULTS: Longer Tp-e interval, greater Tp-e/QT ratio, and higher aVLR were observed in the highest quartile of uric acid level. The incidence of MA-ECG was gradually increased from lowest to highest quartile of serum uric acid in males (p for trend <0.01). After adjusting traditional cardiovascular risk factors, multivariate analysis revealed that the fourth quartile of serum uric acid level was independently associated with MA-ECG in males [odds ratios (OR) (95% confidence interval): 2.129 (1.376-3.295)] but not in females. Serum uric acid was also associated with abnormal baPWV, when adjusted for atherogenic confounders. Compared with the lowest serum uric acid quartile, the OR (95% confidence interval) of the second, third, and fourth quartile were 1.920 (1.246-2.957), 1.650 (1.064-2.558), and 2.501 (1.600-3.908) in males. CONCLUSION: Among China's She ethnic minority, uric acid level was independently related to arterial stiffness assessed by baPWV in both genders. The evaluation of uric acid level was related to higher cardiovascular risk in males but not in females. PMID- 21646857 TI - Detection of glucose abnormalities in patients with acute coronary heart disease: study of reliable tools in clinical practice. AB - AIM: To investigate the prevalence of glucose abnormalities in patients with acute coronary syndrome and to assess the reliability of certain clinical or analytical variables to predict a pathologic result of oral glucose tolerance test (OGTT) at 3 months from discharge. SUBJECTS AND METHODS: Prospective study of 102 patients admitted to the coronary care units. Patients were classified according to the American Diabetes Association criteria. Three months after discharge, an OGTT was performed to non-diabetic patients. RESULTS: Forty-six (45.1%) patients were identified as diabetic (5 previously undiagnosed) and 56 (54.9%) as non-diabetic. OGTT identified 22% of diabetes, 33% of impaired glucose tolerance, and 45% of normal glucose tolerance. Fasting glucose (r=0.55, p<0.001), glycated hemoglobin (HbA1c) (r=0.46, p<0.001), low HDL cholesterol (HDLc) levels (r=-0.34, p<0.02), waist-hip ratio (r=0.45, p<0.01), high systolic blood pressure (r=0.5, p<0.01), and presence of acute myocardial infarction (r=0.46, p<0.001) at admission resulted significant to predict a pathologic result of OGTT. CONCLUSIONS: Glucose abnormalities are frequent in acute coronary syndrome patients. Certain clinical and analytical markers at admission such as fasting glucose, HbA1c, HDL-c<40 mg/dl, waist-hip ratio, and systolic blood pressure, are useful to recognize patients with a higher predisposition to present a pathologic result in OGTT at 3 months from discharge. PMID- 21646858 TI - Improved glucose-stimulated insulin secretion by intra-islet inhibition of protein-tyrosine phosphatase 1B expression in rats fed a high-fat diet. AB - BACKGROUND: Insulin resistance of pancreatic beta-cell itself may be a potential link between systemic insulin resistance and impaired insulin secretion in Type 2 diabetes. Protein tyrosine phosphatase 1B (PTP1B) dephosphorylates tyrosine residues in insulin receptors (IR) and IR substrate (IRS) proteins, and thereby inhibits insulin signaling. Thus the impact of PTP1B expression on beta-cell insulin pathway may affect insulin secretory function. AIM: The aim of the present study was to investigate the effects of intra-islet inhibition of PTP1B expression on glucose-stimulated insulin secretion and potential mechanisms in rats fed a high-fat diet (HFD). MATERIALS AND METHODS: Twenty 10-week-old Sprague Dawley rats were randomly assigned to a regular diet (RD) or a HFD for 8 weeks. At the end of the 8th week, fasting glucose, fasting insulin concentration and lipid profile were measured and an oral glucose tolerance test was done after 12 h fast. Then islet isolation was performed for static incubation and perifusion. Recombinant adenoviruses containing siPTP1B (Ad-siPTP1B), or siControl (Ad siControl) sequences were constructed using AdEasyTM system. Islets were transfected and then assigned to the Ad-siPTP1B group, the Ad-siControl group, and mock control group. Real-time RT-PCR and Western blot were used to evaluate the expression level of PTP1B. Western blot of glucose transporter 2 (GLUT-2) and glucokinsase were also done to investigate the beta-cell glucose-sensing apparatus. Islets were incubated with Krebs-Ringer bicarbonate containing 2.8 mmol/l glucose then 16.7 mmol/l glucose to evaluate glucose-stimulated insulin secretion (GSIS). Islet perifusion was also performed to evaluate kinetics of insulin release in vitro. RESULTS: HFD rats manifested modest glucose intolerance compared with RD group. And PTP1B expression in isolated islets of rats in the HFD group was higher than that of the RD group. GSIS was impaired in islets of HFD rats (2.3+/-0.5-fold as basal for HFD vs 8.1+/-1.3-fold for RD; p<0.05). Ad siPTP1B treatment resulted in 73% decrease in PTP1B mRNA levels and 61% decrease in PTP1B protein compared with islets treated with Ad-siControl (p<0.05). Simultaneously, PTP1B inhibition resulted in 4.7+/-0.8-fold increase of GSIS from basal (vs 1.9+/-0.1-fold for Ad-siControl, p<0.05). Perifusion showed notable improvement of first-phase insulin secretion by AdsiPTP1B treatment. Significant decrease of both GLUT-2 (by 49.8%) and glucokinase (GCK, by 43.7%) were found in the HFD group when compared with the RD group, while up-regulation of both GLUT-2 (by 98%) and GCK (by 62%) was achieved after PTP1B inhibiton by Ad-siPTP1B. CONCLUSIONS: Intra-islet PTP1B is an important physiological regulator of glucose induced insulin release and the characteristics of PTP1B inhibitors in insulin secretion could make it a potential novel therapeutics for protection of beta cell secretory function in Type 2 diabetes. PMID- 21646859 TI - Effect of thyroxine on SNARE complex and synaptotagmin-1 expression in the prefrontal cortex of rats with adult-onset hypothyroidism. AB - Thyroid hormone insufficiency in adulthood causes a wide range of brain impairments, including altered synaptic proteins in the prefrontal cortex (PFC). The present study investigated whether adult-onset hypothyroidism altered the expression of soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexes and synaptotagmin-1 (syt-1) in the PFC of rats. Sprague-Dawley rats were randomly divided into 4 groups: control, hypothyroid, and hypothyroid treated with T(4) [5 or 20 MUg/100 g body weight (BW)]. Adult onset hypothyroidism was induced in rats with the antithyroid drug 6-n-propyl-2 thiouracil (ip injection). PFC levels of synaptosomal-associated protein of 25 kDa (SNAP-25), syntaxin-1, vesicle-associated membrane protein 2 (VAMP-2) and syt 1 were determined by immunohistochemistry and western blot analyses. The results showed that syntaxin-1 and syt-1 were expressed at significantly lower levels in hypothyroid rats, VAMP-2 levels were not altered, and SNAP-25 levels were much higher compared to controls. A 2-week treatment with 5 MUg T(4)/100 g BW partially normalized levels of SNARE complex and syt-1, and 20 MUg T(4)/100 g BW restored these proteins closer to normal levels. Our findings indicate that dysregulation of SNARE complex and syt-1 in PFC of adult-onset hypothyroidism can be restored by T(4) treatment. PMID- 21646860 TI - Remodeling is at the heart of chromatin: the heartaches of chromatin. AB - Chromatin modifications are integral elements of chromosome structure and its function and the vasculature depends on tissue-specific genome regulation for its development. A general concept for the de-regulation of chromatin modifications in cardiac and vascular disease is also emerging. The recognition that metabolic memory contributes to disease persistence highlights the benefit of early and aggressive treatment. As for the importance of memory, we do know that good metabolic control delays the onset of long-term diabetic complications. There are striking parallels between the timing of disease and the development of complications. Landmark multicenter clinical trials on diabetes patients have popularized the concept that glucose is also a demonstrable determinant for the development of complications, indicating the prolonged benefit of intensive therapy and the lasting damage of conventional therapy. Each cell type experiences thousands of modifications to the epigenome in response to environmental changes it is exposed to. Therefore, history is neither lost nor forgotten and previous experiences and exposure may form future memories. There is now a strong resurgence in research trying to understand gene-environment interactions and to determine what commits specific vascular cell types to specific memories. Recent insights show that cardiac gene expression is distinguished by specific chromatin remodeling events and histone modifications that are associated with heart disease. PMID- 21646861 TI - Photodegradation illuminates the role of polyanions in prion infectivity. AB - Understanding the mechanism by which prion infectivity is encoded by the misfolded protein PrP (Sc ) remains a high priority within the prion field. Work from several groups has indicated cellular cofactors may be necessary to form infectious prions in vitro. The identity of endogenous prion conversion cofactors is currently unknown, but may include polyanions and/or lipid molecules. In a recent study, we manufactured infectious hamster prions containing purified PrP (Sc) , co-purified lipid, and a synthetic photocleavable polyanion. The polyanion was incorporated into infectious PrP (Sc) complexes, and then specifically degraded by exposure to ultraviolet light. Light-induced in situ degradation of the incorporated polyanion had no effect on the specific infectivity of the samples as determined by end-point dilution sPMCA and scrapie incubation time assays. Furthermore, prion strain properties were not changed by polyanion degradation, suggesting that intact polyanions are not required to maintain the infectious properties of hamster prions. Here, we review these results and discuss the potential roles cofactors might play in encoding prion infectivity and/or strain properties. PMID- 21646862 TI - Beclin 1-independent autophagy contributes to apoptosis in cortical neurons. AB - Neuronal autophagy is enhanced in many neurological conditions, such as cerebral ischemia and traumatic brain injury, but its role in associated neuronal death is controversial, especially under conditions of apoptosis. We therefore investigated the role of autophagy in the apoptosis of primary cortical neurons treated with the widely used and potent pro-apoptotic agent, staurosporine (STS). Even before apoptosis, STS enhanced autophagic flux, as shown by increases in autophagosomal (LC3-II level, LC3 punctate labeling) and lysosomal (cathepsin D, LAMP1, acid phosphatase, beta-hexasominidase) markers. Inhibition of autophagy by 3-methyladenine, or by lentivirally-delivered shRNAs against Atg5 and Atg7, strongly reduced the STS-induced activation of caspase-3 and nuclear translocation of AIF, and gave partial protection against neuronal death. Pan caspase inhibition with Q-VD-OPH likewise protected partially against neuronal death, but failed to affect autophagy. Combined inhibition of both autophagy and caspases gave strong synergistic neuroprotection. The autophagy contributing to apoptosis was Beclin 1-independent, as shown by the fact that Beclin 1 knockdown failed to reduce it but efficiently reduced rapamycin-induced autophagy. Moreover the Beclin 1 knockdown sensitized neurons to STS-induced apoptosis, indicating a cytoprotective role of Beclin 1 in cortical neurons. Caspase-3 activation and pyknosis induced by two other pro-apoptotic stimuli, MK801 and etoposide, were likewise found to be associated with Beclin 1-independent autophagy and reduced by the knockdown of Atg7 but not Beclin 1. In conclusion, Beclin 1-independent autophagy is an important contributor to both the caspase-dependent and independent components of neuronal apoptosis and may be considered as an important therapeutic target in neural conditions involving apoptosis. PMID- 21646864 TI - Autophagy protects breast cancer cells from epirubicin-induced apoptosis and facilitates epirubicin-resistance development. AB - Epirubicin (EPI) is one of the most effective drugs against cancer. But the acquired resistance of cancer cells to EPI is becoming a major obstacle for successful cancer therapy. Recently, some studies have revealed that macroautophagy (here referred to as autophagy) may protect the cancer cell from anticancer drug-induced death, so autophagy might be related to the development of drug resistance to these reagents. However, the relationship between autophagy and drug resistance has yet to be defined. Our study showed that EPI induced autophagy in human breast cancer MCF-7 cells. And the EPI-induced autophagy protected MCF-7 cells from EPI-induced apoptosis. Furthermore, autophagy was elevated in EPI-resistant MCF-7 cells (MCF-7er cells), and inhibition of autophagy restored the sensitivity of MCF-7er cells to EPI. Therefore, autophagy is a prosurvival factor and has a role in the development of EPI-acquired resistance in EPI-treated MCF-7 cells. Also, this finding indicates that the use of clinically applicable autophagy inhibitors might be one of the important strategies for breast cancer therapy. PMID- 21646863 TI - Src family kinases and paclitaxel sensitivity. AB - Src-family Kinases (SFKs) participate in the regulation of proliferation, differentiation, apoptosis, autophagy, adhesion, migration, invasion and angiogenesis in normal and cancer cells. Abnormal expression of SFKs has been documented in cancers that arise in breast, colon, ovary, melanocyte, gastric mucosa, head and neck, pancreas, lung, and brain. Targeting SFKs in cancer cells has been shown to be a promising therapeutic strategy in solid tumors, particularly in ovarian, colon and breast cancers. Paclitaxel is one of most widely used chemotherapeutic agents for the management of ovarian, breast, lung and head/neck cancers. As a microtubule-stabilizing agent, paclitaxel possesses both mitosis-dependent and mitosis-independent activities against cancer cells. A variety of mechanisms such as deregulation of P-glycoprotein, alteration of tubulin isotypes, alteration of microtubule-regulatory proteins, deregulation of apoptotic signaling pathways, mutation of tubulins and overexpression of copper transporters have been implicated in the development of primary or secondary resistance to paclitaxel. By affecting cancer cell survival, proliferation, autophagy, microtubule stability, motility, and/or angiogenesis, SFKs interact with mechanisms that regulate paclitaxel sensitivity. Inhibition of SFKs can potentiate the anti-tumor activity of paclitaxel by enhancing apoptosis, autophagy and microtubule stability. Based on pre-clinical observations, administration of SFK inhibitors in combination with paclitaxel could improve treatment for ovarian, breast, lung and head/neck cancers. Identification and validation of predictive biomarkers could also permit personalization of the therapy. PMID- 21646865 TI - Health claims substantiation for probiotic and prebiotic products. AB - The topic of "Health Claims Substantiation for Probiotic and Prebiotic Products" was discussed at the 8 (th) annual International Scientific Association for Probiotics and Prebiotics (ISAPP) meeting. The topic is especially timely considering that the regulatory review process for health benefit claims on probiotic and prebiotic products in Europe has not resulted in a single claim being approved (120 negative opinions on probiotic claims and 19 negative opinions on prebiotic claims through February 2011). This situation in Europe and elsewhere has driven companies to seek clarity on a research path that would stand up to scientific scrutiny as well as satisfy regulatory demands for health claim substantiation. It can be challenging to negotiate rigid regulatory distinctions, such as between health and disease, when these states are more realistically represented by continua. One research approach focused on improved homeostasis is explored as a statistically robust approach to measuring physiological parameters in healthy populations, which are the required target for food and supplement claims. Diverse global regulatory frameworks complicate this issue, and harmonization of different approaches globally would simplify requirements for industry, decrease consumer confusion and improve the scientific framework for the research community to set up appropriate research pathways. This report highlights key points from this discussion. PMID- 21646866 TI - Microautophagy in mammalian cells: revisiting a 40-year-old conundrum. AB - The term microautophagy was first used in 1966 by de Duve and Wattiaux and subsequently applied, over the following two decades, to processes described in mammalian cells and involving the presence of lysosome-like organelles having multiple vesicles trapped in their lumen ("multivesicular lysosomes"). Concurrently, many studies suggested a view of microautophagy where the lysosomal membrane was either invaginated or projected arm-like protrusions to sequester cytosolic constituents into intralysosomal vesicles. Although microautophagy in mammalian cells has been traditionally considered as a form of autophagy constitutively active in the turnover of long-lived proteins, little is known about the mechanism and regulation of cargo selection. The lack of specific approaches to directly detect microautophagy in mammalian systems, aside from electron microscopy, is the major current limitation to addressing its physiological role(s) and possible contribution to particular disease states. In this review we consider the current state of knowledge about microautophagic processes. We examine some of the main characteristics of microautophagy in yeast with a view to assessing their relevance for our understanding of microautophagy in mammalian cells. PMID- 21646867 TI - The remarkable capacity for gut microbial and host interactions. AB - The stunning complexity of the resident microbiota and the intricate pathways of microbial and host interactions provide a massive adaptive capacity for mammals. In this addendum we reflect on our recent publication on Toll-like receptor 2 deficiency related colonic mucosal epigenetic, immunologic and microbiomic changes. Our findings underscored the tremendous flexibility of the gut and its microbiota. This flexibility can provide means to overcome significant environmental or genetic challenges. In the meantime, the challenged intestinal system may become vulnerable to otherwise tolerable insults. In such instances, the fine-tuned mutualistic balance between the gut and its microflora may collapse leading to dysbiosis and disease. The ultimate challenge for biomedical research in these cases is to find optimal means for the restoration and maintenance of healthy host physiology. PMID- 21646869 TI - Induction of karyopherin alpha1 expression by indole-3-acetic acid in auxin treated or overproducing tobacco plants. AB - Macromolecules may transfer between the cytoplasm and the nucleus only through specific gates - the nuclear pore complexes (NPCs). Translocation of nucleic acids and large proteins requires the presence of a nuclear localization signal (NLS) within the transported molecule. This NLS is recognized by a class of soluble transport receptors termed karyopherins alpha and beta. We previously characterized the expression pattern of the tomato karyopherin alpha 1 (LeKAPalpha1) promoter in transformed tobacco plants. Expression of LeKAPalpha1 was mainly observed in growing tissues where cell division and extension is rapid. The expression pattern of LeKAPalpha1 resembled that of auxin-responsive genes. This led us to suggest that auxin participates in the regulation of LeKAPalpha1 expression. Here we characterized the correlation between auxin level and the activity of the LeKAPalpha1 promoter. To this end, transgenic tobacco plants carrying the GUS reporter gene under the control of the LeKAPalpha1 promoter were treated with various levels of exogenous auxin. We also studied transgenic plants in which we increased the endogenous levels of auxin. For this, we expressed in plants both the LeKAPalpha1 promoter-GUS reporter and the Agrobacterium tumefaciens iaaM gene, which increases the endogenous levels of auxin. The results indicate that the auxin indole-3-acetic acid (IAA) can induce LeKAPalpha1 expression. We also identified that the sites and levels of LeKAPalpha1 expression correlated with the endogenous pathways of polar auxin transport. PMID- 21646868 TI - Matrix remodeling stimulates stromal autophagy, "fueling" cancer cell mitochondrial metabolism and metastasis. AB - We have previously demonstrated that loss of stromal caveolin-1 (Cav-1) in cancer associated fibroblasts is a strong and independent predictor of poor clinical outcome in human breast cancer patients. However, the signaling mechanism(s) by which Cav-1 downregulation leads to this tumor-promoting microenvironment are not well understood. To address this issue, we performed an unbiased comparative proteomic analysis of wild-type (WT) and Cav-1(-/-) null mammary stromal fibroblasts (MSFs). Our results show that plasminogen activator inhibitor type 1 and type 2 (PAI-1 and PAI-2) expression is significantly increased in Cav-1(-/-) MSFs. To establish a direct cause-effect relationship, we next generated immortalized human fibroblast lines stably overexpressing either PAI-1 or PAI-2. Importantly, PAI-1/2(+) fibroblasts promote the growth of MDA-MB-231 tumors (a human breast cancer cell line) in a murine xenograft model, without any increases in angiogenesis. Similarly, PAI-1/2(+) fibroblasts stimulate experimental metastasis of MDA-MB-231 cells using an in vivo lung colonization assay. Further mechanistic studies revealed that fibroblasts overexpressing PAI-1 or PAI-2 display increased autophagy ("self-eating") and are sufficient to induce mitochondrial biogenesis/activity in adjacent cancer cells, in co-culture experiments. In xenografts, PAI-1/2(+) fibroblasts significantly reduce the apoptosis of MDA-MB-231 tumor cells. The current study provides further support for the "Autophagic Tumor Stroma Model of Cancer" and identifies a novel "extracellular matrix"-based signaling mechanism, by which a loss of stromal Cav 1 generates a metastatic phenotype. Thus, the secretion and remodeling of extracellular matrix components (such as PAI-1/2) can directly regulate both (1) autophagy in stromal fibroblasts and (2) epithelial tumor cell mitochondrial metabolism. PMID- 21646870 TI - Optimization of radiation dose reduction in cardiac computed tomographic angiography. AB - Cardiac computed tomographic angiography (CCTA) has evolved at an unprecedented pace over the past decade, during which time it has proven to be an accurate and effective tool for imaging of the heart in a growing list of clinical applications. However, the rapid growth in the use of CT imaging in general has prompted appropriate concerns regarding increasing medical radiation exposure to patients, particularly with regard to potential long-term risks of radiation induced malignancy on both individual and population levels. As with all medical imaging modalities, imaging the heart with CCTA should be performed in a manner that achieves diagnostic image quality while maintaining patient radiation exposure as low as reasonably achievable (As Low As Reasonably Achievable [ALARA] principle). The goal of this article is to provide the reader with a wide-ranging review of both primary and secondary techniques that are currently available to minimize patient radiation exposure. Some of the techniques described in this article are universal, whereas others may be scanner specific. By gaining a thorough understanding of the various tools and methodologies employed for reduction of radiation exposure, the cardiac imager should be able to formulate CCTA protocols appropriate for their equipment and their clinical applications, in a manner that optimally preserves diagnostic image quality and minimizes patient radiation dose. PMID- 21646871 TI - Revascularization of left main coronary artery disease. AB - Coronary artery disease with left main stenosis is associated with the highest mortality of any coronary lesion. Studies in the 1970s and 1980s comparing coronary artery bypass grafting (CABG) and medical therapy showed a significant survival benefit with revascularization. In the angioplasty era, initial experience with percutaneous intervention was associated with poor clinical outcomes. As a result, percutaneous coronary intervention (PCI) was restricted to patients who were considered inoperable, or those with prior CABG with a functional graft to the left anterior descending or circumflex artery ("protected left main disease"). With the introduction of drug-eluting stents, there are new studies demonstrating comparable survival in patients who were revascularized using PCI and CABG, although percutaneous revascularization is associated with a higher rate of repeat revascularization. In the SYNTAX (Synergy between PCI with Taxus and Cardiac Surgery) trial, the combined incidence of death, myocardial infarction, and stroke was similar between the CABG and PCI groups; however, the stroke rate was higher in the CABG group. The degree and extent of disease as defined by the SYNTAX scoring system has allowed for stratification of risk and improved assignment of patients with left main stenosis to either PCI or CABG. PMID- 21646872 TI - The effects of nonsteroidal anti-inflammatory drugs on blood pressure in hypertensive patients. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are ubiquitous medications used by a wide range of people from otherwise healthy normotensive patients to hypertensive patients with many significant comorbidities. Through a variety of mechanisms related to prostaglandin inhibition, including sodium retention and vasoconstriction, these agents may increase blood pressure. This leads to potentially detrimental effects. A review of the current literature regarding this topic yielded 2 meta-analyses and 10 randomized controlled trials. There is evidence of small blood pressure increases in normotensive patients taking NSAIDs approximating +1.1 mm Hg. Patients with treated hypertension show variable increases with NSAID treatment, ranging up to +14.3 mm Hg for systolic pressure and +2.3 mm Hg for diastolic blood pressure. Most antihypertensive medications seem to have decreased effects with concomitant NSAID administration, with the exception of calcium channel blockers. Given the current literature, it appears that NSAIDs increase blood pressure in patients with controlled-hypertension, but the quantity of this increase is variable. If possible, patients who have hypertension should avoid taking NSAIDs. PMID- 21646873 TI - Vitamin D status and risk of cardiovascular events: lessons learned via systematic review and meta-analysis. AB - Accumulating data linking hypovitaminosis D to cardiovascular (CV) events has contributed to large increases in vitamin D testing and supplementation. To evaluate the merits of this practice, we conducted a systematic review with meta analysis providing a framework for interpreting the literature associating hypovitaminosis D with increased CV events. Prospective studies were identified by search of MEDLINE and EMBASE from inception to January 2010, restricted to English language publications. Two authors independently extracted data and graded study quality. Pooled relative risks (RR) were calculated using a random effects model. Ten studies met criteria for review and 7 were included in meta analysis. Pooled RR for CV events using FAIR and GOOD quality studies was 1.67 (95% confidence interval, 1.23-2.28) during an average follow-up of 11.8 years. There was evidence of significant heterogeneity across studies (Q statistics = 16.6, P = 0.01, I = 63.8%), which was eliminated after omitting 2 studies identified by sensitivity analysis (RR, 1.34 [1.08-1.67]; P for heterogeneity =0.33). When restricting analysis to GOOD quality studies (RR, 1.27 [1.04-1.56]), no significant heterogeneity was found (P = 0.602). Systematic review identified significant shortcomings in the literature, including variability in defining vitamin D status, seasonal adjustments, defining and determining CV outcomes, and the use of baseline vitamin D levels. In conclusion, a modest increased risk of CV events associated with hypovitaminosis D is tempered by significant limitations within the current literature. These findings underscore the importance of critical appraisal of the literature, looking beyond reported risk estimates before translating results into clinical practice. PMID- 21646874 TI - Cardiovascular disease in US firefighters: a systematic review. AB - Cardiovascular disease (CVD) is the leading cause of on-duty death among firefighters (45% of on-duty fatalities) and a major cause of morbidity. CVD in the fire service also has adverse public safety implications as well as significant cost impacts on government agencies. Over the last decade, our understanding of CVD among firefighters has significantly improved and provides insight into potential preventive strategies. The physiology of cardiovascular arousal and other changes that occur in association with acute firefighting activities have been well-characterized. However, despite the strenuous nature of emergency duty, firefighters' prevalence of low fitness, obesity, and other CVD risk factors are high. Unique statistical approaches have documented that on-duty CVD events do not occur at random in the fire service. They are more frequent at certain times of day, certain periods of the year, and are overwhelmingly more frequent during strenuous duties compared with nonemergency situations. Moreover, as expected on-duty CVD events occur almost exclusively among susceptible firefighters with underlying CVD. These findings suggest that preventive measures with proven benefits be applied aggressively to firefighters. Furthermore, all fire departments should have entry-level medical evaluations, institute periodic medical and fitness evaluations, and require rigorous return to work evaluations after any significant illness. Finally, on the basis of the overwhelming evidence supporting markedly higher relative risks of on-duty death and disability among firefighters with established coronary heart disease, most firefighters with clinically significant coronary heart disease should be restricted from participating in strenuous emergency duties. PMID- 21646875 TI - 3-methylglutaconic aciduria type IV: a syndrome with an evolving phenotype. PMID- 21646877 TI - Genotypic resistance testing in routine clinical care. AB - PURPOSE OF REVIEW: Genotypic resistance testing has become part of routine clinical management of HIV-infected patients. Focussing on observational studies, this review looks at recent advances in this area. RECENT FINDINGS: Translation of the nucleotide sequence generated by the resistance test into clinically useful information remains a major challenge. A recent key development is the availability of therapy optimization tools to predict regimens that are most likely to achieve virological suppression. Standard genotypic resistance testing only examines protease and part of reverse transcriptase; as drugs are licensed to further targets, it has become necessary to expand the repertoire for testing. Traditionally, genotypic testing has not been attempted at viral loads less than 1000 copies/ml, but recent studies indicate that major mutations are often detected at much lower levels. Similarly, various methods have been developed for the detection of minority variants including allele-specific PCR, single-genome sequencing, and ultra-deep sequencing. SUMMARY: The technology and interpretation of genotypic resistance tests is in a phase of rapid development. It remains uncertain which of these developments will become part of routine clinical practice. PMID- 21646879 TI - Bibliography. Cohort analysis of clinical and treatment outcomes. Current world literature. PMID- 21646878 TI - HIV-1 treatment as prevention: the good, the bad, and the challenges. AB - PURPOSE OF REVIEW: This work focuses on the use of antiretroviral agents to prevent the sexual transmission of HIV-1. RECENT FINDINGS: Two randomized clinical trials demonstrated that antiretroviral agents provided before exposure to HIV-1 offer substantial protection, ostensibly directly proportional to the concentration of antiretroviral therapy (ART) in the genital secretions. Intense focus on the use of HIV treatment as prevention has led to publication of modeling exercises, ecological studies, and observational studies, most of which support the potential benefits of ART. However, the logistical requirements for successful use of ART for prevention are considerable. SUMMARY: ART will serve as a cornerstone of combination prevention of HIV-1. Continued research will be essential to measure anticipated benefits and to detect implementation barriers and untoward consequences of such a program, especially increases in primary ART resistance. PMID- 21646880 TI - High-performance work systems in health care management, part 1: development of an evidence-informed model. AB - BACKGROUND: : Although management practices are recognized as important factors in improving health care quality and efficiency, most research thus far has focused on individual practices, ignoring or underspecifying the contexts within which these practices are operating. Research from other industries, which has increasingly focused on systems rather than individual practices, has yielded results that may benefit health services management. PURPOSE: : Our goal was to develop a conceptual model on the basis of prior research from health care as well as other industries that could be used to inform important contextual considerations within health care. METHODOLOGY/APPROACH: : Using theoretical frameworks from A. Donabedian (1966), P. M. Wright, T. M. Gardner, and L. M. Moynihan (2003), and B. Schneider, D. B. Smith, and H. W. Goldstein (2000) and review methods adapted from R. Pawson (2006b), we reviewed relevant research from peer-reviewed and other industry-relevant sources to inform our model. The model we developed was then reviewed with a panel of practitioners, including experts in quality and human resource management, to assess the applicability of the model to health care settings. FINDINGS: : The resulting conceptual model identified four practice bundles, comprising 14 management practices as well as nine factors influencing adoption and perceived sustainability of these practices. The mechanisms by which these practices influence care outcomes are illustrated using the example of hospital-acquired infections. In addition, limitations of the current evidence base are discussed, and an agenda for future research in health care settings is outlined. PRACTICE IMPLICATIONS: : Results may help practitioners better conceptualize management practices as part of a broader system of work practices. This may, in turn, help practitioners to prioritize management improvement efforts more systematically. PMID- 21646881 TI - High-performance work systems in health care management, part 2: qualitative evidence from five case studies. AB - BACKGROUND: : A capable workforce is central to the delivery of high-quality care. Research from other industries suggests that the methodical use of evidence based management practices (also known as high-performance work practices [HPWPs]), such as systematic personnel selection and incentive compensation, serves to attract and retain well-qualified health care staff and that HPWPs may represent an important and underutilized strategy for improving quality of care and patient safety. PURPOSE: : The aims of this study were to improve our understanding about the use of HPWPs in health care organizations and to learn about their contribution to quality of care and patient safety improvements. METHODOLOGY/APPROACH: : Guided by a model of HPWPs developed through an extensive literature review and synthesis, we conducted a series of interviews with key informants from five U.S. health care organizations that had been identified based on their exemplary use of HPWPs. We sought to explore the applicability of our model and learn whether and how HPWPs were related to quality and safety. All interviews were recorded, transcribed, and subjected to qualitative analysis. FINDINGS: : In each of the five organizations, we found emphasis on all four HPWP subsystems in our conceptual model-engagement, staff acquisition/development, frontline empowerment, and leadership alignment/development. Although some HPWPs were common, there were also practices that were distinctive to a single organization. Our informants reported links between HPWPs and employee outcomes (e.g., turnover and higher satisfaction/engagement) and indicated that HPWPs made important contributions to system- and organization-level outcomes (e.g., improved recruitment, improved ability to address safety concerns, and lower turnover). PRACTICE IMPLICATIONS: : These case studies suggest that the systematic use of HPWPs may improve performance in health care organizations and provide examples of how HPWPs can impact quality and safety in health care. Further research is needed to specify which HPWPs and systems are of greatest potential for health care management. PMID- 21646882 TI - Talent management best practices: how exemplary health care organizations create value in a down economy. AB - BACKGROUND: : Difficult economic conditions and powerful workforce trends pose significant challenges to managing talent in health care organizations. Although robust research evidence supports the many benefits of maintaining a strong commitment to talent management practices despite these challenges, many organizations compound the problem by resorting to workforce reductions and limiting or eliminating investments in talent management. PURPOSE: : This study examines how nationwide health care systems address these challenges through best practice talent management systems. Addressing important gaps in talent management theory and practice, this study develops a best practice model of talent management that is grounded in the contextual challenges facing health care practitioners. METHODOLOGY: : Utilizing a qualitative case study that examined 15 nationwide health care systems, data were collected through semistructured interviews with 30 executives and document analysis of talent management program materials submitted by each organization. FINDINGS: : Exemplary health care organizations employ a multiphased talent management system composed of six sequential phases and associated success factors that drive effective implementation. Based on these findings, a model of talent management best practices in health care organizations is presented. PRACTICE IMPLICATIONS: : Health care practitioners may utilize the best practice model to assess and enhance their respective talent management systems by establishing the business case for talent management, defining, identifying, and developing high-potential leaders, carefully communicating high-potential designations, and evaluating talent management outcomes. PMID- 21646883 TI - Forcing the system: a configuration analysis of a regionalized neonatal-perinatal health network. AB - BACKGROUND: : Health care transformations often involve the development of networks to ensure smooth and safe patient flows throughout the care continuum. However, more empirical information is needed on the workings of health networks and on how their structures, processes, and systems influence access to high quality patient care. PURPOSE: : Using Miller's concept of configuration, we describe the workings of a health network specialized in the provision of neonatal-perinatal care, a specialty where accessibility issues are quite problematic. We aimed to generate evidence that will assist policy makers, network managers, and clinicians in facilitating access to high-quality neonatal perinatal care. METHODOLOGY/APPROACH: : From late 2007 to early 2008, we conducted a case study of all (N = 7) neonatal intensive care units (NICUs) in the province of Quebec (Canada). We performed field work into two purposefully selected NICUs. This involved 450 hours of nonparticipant observation and 56 semistructured interviews with various actors. Data from these sources were triangulated with data collected during informal interviews with key actors from the other five NICUs in the province and from administrative databases. FINDINGS: : We found that the elements of this health network are pulled together by a core orchestrating theme: "Forcing the system." Indeed, in attempting to fulfill the network mission of providing access to high-quality neonatal-perinatal care, clinicians and managers must implement various strategies to compensate for the misfit of the configuration. Although these strategies are successful in providing access to neonatal-perinatal care, they, however, have adverse effects that are paradoxically in contradiction with the network's core mission. PRACTICE IMPLICATIONS: : This configuration analysis enabled us to identify a set of modifiable elements that contribute to the misfit of the configuration and its suboptimal functioning. The comprehensiveness of the configuration approach was proven useful for the analysis of such a complex organizational form. PMID- 21646884 TI - Staff, space, and time as dimensions of organizational slack: a psychometric assessment. AB - BACKGROUND: : In the theoretical and research literature, organizational slack has been largely described in terms of financial resources and its impact on organizational outcomes. However, empirical research is limited by unclear definitions and lack of standardized measures. PURPOSE: : The aim of this study was to assess the psychometric properties of a new organizational slack measure in health care settings. METHODS: : A total of 752 nurses and 197 allied health care professionals (AHCPs) employed in seven pediatric Canadian hospitals completed the Alberta Context Tool, an instrument measuring organizational context, which includes the newly developed organizational slack measure. The nine-item, 5-point Likert organizational slack measure includes items assessing staff perceptions of available human resources (staffing), time, and space. We report psychometric assessments, bivariate analyses, and data aggregation indices for the measure. FINDINGS: : The findings indicate that the measure has three subscales (staff, space, and time) with acceptable internal consistency reliability (alphas for staff, space, and time, respectively:.83,.63, and.74 for nurses;.81,.52, and.76 for AHCPs), links theory and hypotheses (construct validity), and is related to other relevant variables. Within-group reliability measures indicate stronger agreement among nurses than AHCPs, more reliable aggregation results in all three subscales at the unit versus facility level, and higher explained variance and validity of aggregated scores at the unit level. PRACTICE IMPLICATIONS: : The proposed organizational slack measure assesses modifiable organizational factors in hospitals and has the potential to explain variance in important health care system outcomes. Further assessments of the psychometric properties of the organizational slack measure in acute and long term care facilities are underway. PMID- 21646885 TI - Experiences with electronic health records: early adopters in long-term care facilities. AB - BACKGROUND: : Electronic health records (EHRs) are becoming a required technology across the health care sector. Long-term care (LTC) facilities have lagged other settings in adopting health information technologies but represent an area where significant care coordination benefits might be realized. Nevertheless, managers face many of the same challenges implementing EHRs that exist in other environments when implementing enterprise-wide systems. PURPOSES: : This study was conducted to provide a description of the early users' experiences with EHRs in LTC facilities. METHODOLOGY: : Semistructured interviews were conducted. The 10 sites were all the "freestanding" LTC facilities using an EHR as of July 2008 in Texas. The interview respondents included administrators, nursing managers, nurses, certified nurse aides, and other system users. Semistructured interviews across multiple stakeholders were used to assess constructs critical to EHR adoption and implementation. FINDINGS: : The LTC facility employees who work with EHR systems on a daily basis were positive about their experiences. In particular, operational improvements were achieved through increased access to resident information, cost avoidance, increased documentation accuracy, and implementation of evidence-based practices. PRACTICE IMPLICATIONS: : Overall, administrators believed that the systems improved care quality and employee satisfaction and were cost effective and that the EHR made a positive return on investment. Electronic documentation led to both increases in charge capture related to resource utilization group documentation, significant savings in pharmacy waste, and reductions in nursing overtime as medical record management became more automated. Quality improvement came from computer-aided monitoring of the certified nurse aide's attendance to residents' activities of daily living. PMID- 21646886 TI - Environmental factors and health information technology management strategy. AB - BACKGROUND: : Previous studies have provided theoretical and empirical evidence that environmental forces influence hospital strategy. PURPOSES: : Rooted in resource dependence theory and the information uncertainty perspective, this study examined the relationship between environmental market characteristics and hospitals' selection of a health information technology (HIT) management strategy. METHODOLOGY/APPROACH: : A cross-sectional design is used to analyze secondary data from the American Hospital Association Annual Survey, the Healthcare Information and Management Systems Society Analytics Database, and the Area Resource File. Univariate and multinomial logistic regression analyses are used. FINDINGS: : Overall, 3,221 hospitals were studied, of which 60.9% pursed a single-vendor HIT management strategy, 28.9% pursued a best-of-suite strategy, and 10.2% used a best-of-breed strategy. Multivariate analyses controlling for hospital characteristics found that measures of environmental factors representing munificence, dynamism, and/or complexity were systematically associated with various hospital HIT management strategy use. Specifically, the number of generalist physicians per capita was positively associated with the single-vendor strategy (B = -5.64, p = .10). Hospitals in urban markets were more likely to pursue the best-of-suite strategy (B = 0.622, p < .001). Dynamism, measured as the number of managed care contracts for a given hospital, was negatively associated with the single-vendor strategy (B = 0.004, p = .049). Lastly, complexity, measured as market competition, was positively associated with the best-of-breed strategy (B = 0.623, p = .042). PRACTICE IMPLICATIONS: : By and large, environmental factors are associated with hospital HIT management strategies in mostly theoretically supported ways. Hospital leaders and policy makers interested in influencing the adoption of hospital HIT should consider how market conditions influence HIT management decisions as part of programs to promote meaningful use. PMID- 21646888 TI - Nothing is permanent but change (Heraclitus). PMID- 21646889 TI - Benchmarks of case management: ingenuity, creativity, and tenacity--stories from the British Columbia Interior, Canada. PMID- 21646890 TI - Understanding a new order for care delivery. PMID- 21646891 TI - Reflective practice: a framework for case manager development. AB - PURPOSE/OBJECTIVES: The role of a nurse case manager (NCM) incorporates practice that is built upon knowledge gained in other roles as well as components unique to case management. The concept of reflective practice was used in creating a framework to recognize the developmental stages that occur within community based case management practice. The formation of this framework and its uses are described in this article. PRIMARY PRACTICE SETTING: The practice setting is a community based case management department in a large midwestern metropolitan health care system with Magnet recognition. Advanced practice nurses provide care for clients with chronic health conditions. FINDINGS/CONCLUSIONS: Twenty-four narratives were used to identify behaviors of community based case managers and to distinguish stages of practice. The behaviors of advanced practice found within the narratives were labeled and analyzed for similarities. Related behaviors were grouped and descriptor statements were written. These statements grouped into 3 domains of practice: relationship/partnership, coordination/collaboration, and clinical knowledge/decision making. The statements in each domain showed practice variations from competent to expert, and 3 stages were determined. Reliability and validity of the framework involved analysis of additional narratives. The reflective practice process, used for monthly case review presentations, provides opportunity for professional development and group learning focused on improving case manager practice. The framework is also being used in orientation as new case managers acclimate to the role. IMPLICATIONS FOR CASE MANAGEMENT PRACTICE: Reflective writing has unveiled the richness and depth of nurse case manager practice. The depth of knowledge and skills involved in community-based case management is captured within this reflective practice framework. This framework provides a format for describing community based case manager practice development over the course of time and has been used as a tool for orientation and peer review. PMID- 21646893 TI - Role ambiguity and role conflict in nurse case managers: an integrative review. AB - PURPOSE/OBJECTIVES: The purpose of this integrative review is to critically examine the relationship between the transition from a direct caregiver to a nurse case manager role and the perceived levels of role ambiguity and role conflict. PRIMARY PRACTICE SETTING(S): Nurse case managers in acute care, postacute care, and managed care settings. FINDINGS/CONCLUSIONS: Nurses can expect to experience substantial role ambiguity and role conflict as they take on the case manager role, primarily because of inadequate role definition, unexpected ethical challenges, and lack of prior insight into the case manager role. Role ambiguity and role conflict may impact nurse case managers' job satisfaction and job performance. IMPLICATIONS FOR CASE MANAGEMENT PRACTICE: Well designed broad-scale descriptive studies are needed to further explicate the relationship between transition from direct caregiver to nurse case manager roles and the experience of role ambiguity and role conflict. Nurses transitioning from direct caregiver to nurse case manager roles should be systematically prepared to identify and manage specific ethical challenges commonly encountered by case managers. Further development of a nursing theoretical foundation for case management would be very useful in helping caregivers understand that as case managers, they remain practicing nurses with all the inherent rights and responsibilities of professional nursing. PMID- 21646895 TI - Case management redesign in an urban facility. AB - PURPOSE OF THE STUDY: To explore strategies for improving patient throughput and to redesign case management processes to facilitate level of care transitions and safe discharges. PRIMARY PRACTICE SETTING: Large Urban Medical Center in South Los Angeles County, with 384 licensed beds that services poor, underserved communities. METHODOLOGY AND SAMPLE: Both qualitative and quantitative methods were applied. Combined theoretical frameworks were used for needs assessment, intervention strategies, and change management. Observations, interviews, surveys, and database extraction methods were used. The sample consisted of case management staff members and several other staff from nursing, social work, and emergency department staff. RESULTS: Postintervention measures indicated improvement in reimbursements for services, reduction in length of stay, increased productivity, improved patients' access to care, and avoiding unnecessary readmission or emergency department visits. IMPLICATIONS FOR CASE MANAGEMENT PRACTICE: Effective change management strategies must consider multiple factors that influence daily operations and service delivery. Creating accountability by using performance measures associated with patient transitions is highlighted by the case study results. The authors developed a process model to assist in identifying and tracking outcome measures related to patient throughput, front-end assessments, and effective patient care transitions. This model can be used in future research to further investigate best case management practices. PMID- 21646896 TI - The silence is deafening. PMID- 21646897 TI - To chat or not to chat? That is the question. PMID- 21646898 TI - 2011 presidential initiative: strengthening our military families. PMID- 21646899 TI - Case management down under. PMID- 21646900 TI - Addressing the "aging out" problem in the work place. PMID- 21646901 TI - Addressing the language barrier with non-English speaking claimants. PMID- 21646902 TI - Relationship between renal dysfunction, nephrotoxicity and death among HIV adults on tenofovir. AB - OBJECTIVE: In April 2010, the South African government added tenofovir disoproxil fumarate to its first-line antiretroviral therapy (ART) for HIV patients. We analyzed the relationship between renal dysfunction at tenofovir initiation, nephrotoxicity and mortality. DESIGN: Cohort analysis of HIV-infected adults who received tenofovir and had a creatinine clearance done at initiation at the Themba Lethu Clinic, Johannesburg, South Africa, between April 2004 and September 2009. METHODS: We estimated the relationship between renal dysfunction, nephrotoxicity [any decline in kidney function from baseline (acute or chronic) that is secondary to a toxin (including drugs)] and mortality for patients initiated onto tenofovir-containing regimens using marginal structural models and inverse probability of treatment weights to correct estimates for lost to follow up and confounding. RESULTS: Of 890 patients initiated onto tenofovir, 573 (64.4%) had normal renal function (>=90 ml/min), 271 (30.4%) had mild renal dysfunction (60-89 ml/min) and 46 (5.2%) had moderate renal dysfunction (30-59 ml/min). A total of 2.4% experienced nephrotoxicity, 7.8% died and 9.7% were lost during 48 months of follow-up. Patients with mild [hazard ratio 4.8; 95% confidence interval (CI) 1.5-15.2] or moderate (hazard ratio 15.0; 95% CI 3.4 66.5) renal dysfunction were at greatest risk of nephrotoxicity, whereas those with mild (hazard ratio 1.2; 95% CI 0.7-2.3) or moderate (hazard ratio 3.2; 95% CI 1.3-7.8) renal dysfunction vs. normal renal function were at highest risk of death by 48 months. CONCLUSION: Much of the incident renal dysfunction in tenofovir patients is likely related to preexisting renal disorder, which may be exacerbated by tenofovir. With expanded use of tenofovir, screening for renal dysfunction prior to initiation and dose adjustment is necessary to help improve ART outcomes. PMID- 21646903 TI - Risk of cerebrovascular events in persons with and without HIV: a Danish nationwide population-based cohort study. AB - OBJECTIVE: To assess the risk of cerebrovascular events (CVEs) in HIV-infected individuals and evaluate the impact of proven risk factors, injection drug abuse (IDU), immunodeficiency, HAART and family-related risk factors. DESIGN: Nationwide, population-based cohort study. METHODS: The study population included all Danish HIV-infected individuals, a population-based comparison cohort and parents of both cohorts - all with no prior cerebral comorbidity. We computed incidence rate ratios (IRRs) of overall CVEs and CVEs with and without proven risk factors, stratifying the analyses on IDU. Impact of immunodeficiency, HAART, protease inhibitors, indinavir, didanosin, tenofovir and abacavir on risk of CVEs was analyzed using time-dependent Cox regression analyses. RESULTS: HIV-infected individuals had an increased risk of CVEs compared with the comparison cohorts [(non-IDU HIV adjusted IRR 1.60; 95% confidence interval [CI] 1.32-1.94), (IDU HIV adjusted IRR 3.94; 95% CI 2.16-7.16)]. The risk was increased with and without proven risk factors. A CD4 cell count of 200 cells/MUl or less before the start of HAART and exposure to abacavir increased the risk of CVE [(adjusted IRR 2.26; 95% CI 1.05-4.86) and (adjusted IRR 1.66; 95% CI 1.03-2.68)]. Protease inhibitors, indinavir, didanosin, tenofovir and HAART in general had no impact. Risk of CVEs was only increased in the parents of IDU HIV-infected individuals. CONCLUSION: HIV-infected individuals have an increased risk of CVEs with and without proven risk factors. The risk is associated with IDU, low CD4 cell count and exposure to abacavir, but not with HAART. An association with family-related risk factors seems vague except for parents of IDU individuals. PMID- 21646904 TI - Homicide due to intravenous metallic mercury injection followed by sodium cyanide injection. AB - We report a case of homicide due to intravenous mercury injection followed by meperidine and sodium cyanide injection. A 35-year-old woman was found dead in bed at home by her husband. Reportedly, she had been sick for more than 5 months. Initial death investigation revealed no evidence of foul play. Her death was believed to be natural. Therefore, her body was buried without an autopsy. Two months after death, her family requested an autopsy because they suspected her physician husband killed her. Her body was exhumed, and an autopsy was performed. Postmortem examination revealed numerous metallic mercury globules in the pulmonary arteries. Toxicological analysis revealed a high concentration of mercury in the tissue samples of the lungs, liver, heart, and kidney. In addition, cyanide and meperidine were also found in the heart and liver. The detailed case history and postmortem examination findings are described. PMID- 21646905 TI - Tattoo frequency and types among homicides and other deaths, 2007-2008: a matched case-control study. AB - An estimated 25% of the US population aged 18 to 50 years has a tattoo, which have been associated with markers of high-risk behaviors including alcohol and drug use, violence, carrying weapons, sexual activity, eating disorders, and suicide. This study compares tattoo prevalence and type in a homicide population to those of an age-, race-, and sex-matched control group of nonhomicide deaths. The data for this study were abstracted from autopsy records maintained by the Jefferson County Alabama Coroner/Medical Examiner's Office for the years 2007 and 2008. Odds ratios and 95% confidence intervals for the association between homicide and tattoo presence and characteristics were calculated using conditional logistic regression. There was no association between tattoo presence and death by homicide; however, among blacks, memorial tattoos were significantly more common among homicides compared with other types of deaths (odds ratio, 2.50; 95% confidence interval, 1.10-5.68). The results of the current study suggest that specific types of tattoos, but not all tattoos, may be risk factors for homicide. Other factors, such as race and lifestyle, along with tattoos may need to be considered. PMID- 21646906 TI - Bioimpedance and the duration of the hemodialysis session. AB - Bioelectrical impedance analysis (BIA) is composed of resistance (R) and reactance (Xc). The aim of this study was to investigate whether BIA may be influenced by the duration of hemodialysis (HD) sessions. Eleven uremic patients underwent one 4-hour and one 8-hour bicarbonate HD session. Volume of blood and dialysate processed, volume of ultrafiltration (V(UF)), and dialysate electrolyte concentrations were prescribed to be the same. R and Xc were determined at the start and the end of each session, injecting 800 MUA at 50 kHz alternating sinusoidal current (BIA 101; Akern, Italy). Mean pre- and postdialysis body weights and V(UF) were not significantly different in the 4-hour and 8-hour treatments. Postdialysis R, DeltaR (the difference between post- and predialysis R values), and percent increase of R values were significantly higher in the 8 hour sessions, when compared with the corresponding values of the 4-hour sessions (p < 0.0001, 0.02, and 0.02, respectively). In conclusion, this study shows that 8-hour HD sessions were associated with postdialysis R, DeltaR, and percent increase of R values significantly higher than the corresponding ones of 4-hour sessions. If higher R values may represent a proxy of a correct dry body weight, it remains a matter of future research. PMID- 21646907 TI - Effect of tapentadol extended release on productivity: results from an analysis combining evidence from multiple sources. AB - OBJECTIVE: To compare the effects of tapentadol-extended release versus oxycodone controlled release for pain relief on productivity by combining evidence from different sources. METHODS: Multiparameter evidence synthesis. Three sources were used. The first consisted of 3 randomized double-blind controlled trials that evaluated the efficacy and safety of tapentadol and oxycodone for the management of chronic pain. The second was published data on the incidence of constipation in patients exposed to opioids, and the third was a published survey that evaluated the effect of opioid-induced constipation on productivity. In the trials, a patient was classified as constipated if constipation was reported at any time during the 15 weeks of double-blinded assessment after randomization. In the survey, the effect of constipation on productivity was measured using the Work Productivity and Activity Impairment Questionnaire. All analyses were performed using Bayesian Markov chain Monte Carlo simulations in WinBUGS. RESULTS: The odds of developing constipation were 60% lower with tapentadol than with oxycodone (odds ratio=0.40, 95% credible interval, 0.32-0.50). Tapentadol was associated with less time missed from work, less impairment while working, and a lower overall loss in work productivity compared with oxycodone. The gain in overall work productivity with tapentadol was 1.92% compared with oxycodone (95% credible interval, 1.32-2.59), which translates to a gain of almost 1 hour per week worked. DISCUSSION: Tapentadol was associated with increases in all productivity dimensions compared with oxycodone. Multiparameter evidence synthesis capitalizes on available evidence, so that better informed medical decisions can be made. PMID- 21646908 TI - Graft rejection after femtosecond laser--assisted deep anterior lamellar keratoplasty: report of 3 cases. AB - PURPOSE: To report 3 cases of stromal rejection after deep anterior lamellar keratoplasty (DALK) assisted by a femtosecond laser. METHODS: Three keratoconus eyes of 2 men (22 and 30 years old) and 1 woman (24 years old) who had DALK with a 60-kHz femtosecond laser developed intrastromal graft rejection with superficial and deep neovascularization after 6, 15, and 12 months, respectively. All patients underwent confocal microscopy and were treated with topical steroid therapy. RESULTS: In vivo confocal microscopy revealed cellular inflammatory infiltrates in the subepithelial and middle stroma of the donor lamella without involvement of the endothelium. Graft rejection was rapidly reversed with topical steroid therapy, and a clear cornea was restored in all cases. CONCLUSIONS: Graft rejection remains a significant complication of lamellar surgery but is associated with good tissue restoration and complete visual recovery. PMID- 21646909 TI - Collaboration between prehospital emergency medical teams and palliative care networks allows a better respect of a patient's will. AB - This study aimed to evaluate whether patient's wishes were respected by prehospital emergency medical teams after implementing collaboration and a standardized process between a community-based palliative network and the Emergency Medical Service system. Forty patients were included. In 75% of cases, the doctor of the prehospital emergency team decided in collaboration with the network's doctor according to the established procedure. This has enabled a respect of the care plan in 83% of cases. Without collaboration with the palliative care network, through the ignorance of its existence or through the wish of the prehospital emergency medical teams for taking decisions alone, the care plan was only respected in 40% of cases, P=0.025. Collaboration between prehospital emergency medical teams and community-based palliative care networks seems to enable a better respect of the care plan in the event of emergency situations affecting the patient in a palliative situation. PMID- 21646910 TI - A randomized controlled trial comparing lactulose, probiotics, and L-ornithine L aspartate in treatment of minimal hepatic encephalopathy. AB - BACKGROUND: Minimal hepatic encephalopathy (MHE) impairs daily functioning and health-related quality of life (HRQoL). The modalities of treatment of MHE have not been adequately studied. AIMS: To compare lactulose, probiotics, and L ornithine L-aspartate (LOLA) in treatment of MHE and effect on HRQoL by Sickness Impact Profile questionnaire. METHODS: Consecutive patients with cirrhosis were screened for MHE. MHE was diagnosed by two or more abnormal psychometric tests (number/figure connection tests A and B, block design test, picture completion test). Patients were randomized to no treatment (GpA), lactulose 30-60 ml/twice per day (GpB), probiotics 110 billion colony forming units twice in a day (GpC), LOLA 6 g three times per day (GpD) for 3 months. Arterial ammonia and HRQoL assessment using SIP questionnaire was done at baseline and at 3 months. RESULTS: One hundred and sixty (49.69%) of 322 patients with cirrhosis had MHE. After 3 months, MHE recovered in GpA four (10%), GpB 19 (47.5%), GpC 14 (35%), and GpD 14 (35%). MHE improved significantly in all three treatment groups (GpB, GpC, GpD) compared with no treatment (GpA) (P=0.006). Overt hepatic encephalopathy developed in nine (5.6%) of 160 patients; GpA four (10%), GpB one (2.5%), GpC two (5%), and GpD two (5%), respectively. There was significant improvement in SIP score in GpB (6.98+/-4.1), GpC (6.24+/-3.4), and GpD (7.33+/-3.8) versus GpA (1.05+/-2.6), P value of less than 0.001. The decrease in SIP score correlated with an improvement in MHE on multivariate analysis but there was no correlation with the type of intervention offered. There was no significant change in arterial ammonia level after therapy in GpA (-0.52+/-7.8 MUmol/l). Arterial ammonia level in GpB (-8.47+/-5.8 MUmol/l), GpC (-7.31+/-7.9 MUmol/l), and GpD ( 9.61+/-9.3 MUmol/l) were significantly more than GpA (P<0.0001). CONCLUSION: Lactulose, probiotics, and LOLA significantly improve MHE and HRQoL in patients with chronic liver disease. PMID- 21646911 TI - HIV-1 infection impairs HSV-specific CD4(+) and CD8(+) T-cell response by reducing Th1 cytokines and CCR5 ligand secretion. AB - BACKGROUND: The concept of HIV-1/HSV-negative immunosynergy has recently come to light, which leads us to explore the impact of HIV-1 infection on HSV-specific T cell immunity. METHODS: : A combination of interferon (IFN)-gamma ELISpot and Luminex-based multicytokine profiling assays was used to compare, in a cross sectional study, the HSV-specific CD4 and CD8 T-cell responses between 20 HIV 1/HSV-coinfected and 12 HIV-1-uninfected/HSV-infected individuals after in vitro restimulation with HSV glycoprotein D (gD) peptide epitopes. RESULTS: In response to CD4 and CD8 gD peptide epitopes, mean value (+/-standard errors of the mean) of the different IFN-gamma-secreting T cells (ISC) means was significantly reduced in HIV-1/HSV-coinfected individuals (70 ISC +/- 10 and 60 ISC +/- 8/10 cells) compared with HIV-1-uninfected/HSV-infected individuals (280 ISC +/- 25 and 234 ISC +/- 23/10 cells, both P < 0.001). After stimulation with the immunodominant CD4 gD and CD8 gD peptide epitopes, the Th1 cytokine and CCR5 ligand secretions were decreased in the HIV-1-infected group although Th17 cytokines increased. The mean concentration of interleukin (IL)-2, IFN-gamma, the IFN-gamma-induced protein 10 kDa, and the monokine induced by IFN-gamma was correlated to the mean concentration of macrophage inflammatory proteins (MIP 1alpha, MIP-1beta), RANTES and Eotaxin (rho = 0.56, P = 0.02 and rho = 0.52, P = 0.03). CONCLUSIONS: HIV-1 infection impairs both the number and function of HSV specific T cells. The downregulation of Th1 cytokines and CCR5 ligands in HIV 1/HSV-coinfected individuals may further facilitate both HSV reactivations and HIV-1 replication. PMID- 21646912 TI - Plasma sCD14 is a biomarker associated with impaired neurocognitive test performance in attention and learning domains in HIV infection. AB - OBJECTIVE: Mild forms of HIV-associated neurocognitive disorders (HAND) remain prevalent in the era of combination antiretroviral therapy (cART). Although elevated lipopolysaccharide (LPS) and immune activation are implicated in HAND pathogenesis, relationships of LPS and inflammatory markers to mild forms of HAND or impairment in specific cognitive domains are unknown. To examine these relationships, we compared plasma soluble CD14 (sCD14), CCL2, and LPS levels with neurocognitive test scores in a cART era cohort. METHODS: We analyzed plasma from HIV+ subjects (n = 97) with nadir CD4 counts <300 and high frequency of hepatitis C virus coinfection and illicit drug use for relationships between sCD14, CCL2, and LPS levels and neurocognitive test scores. RESULTS: Plasma sCD14 levels were higher in subjects with test scores indicating global impairment (P = 0.007), particularly in attention and learning domains (P = 0.015 and P = 0.03, respectively), regardless of HAND diagnosis. Plasma sCD14 levels correlated inversely with global, attention, and learning T scores (P = 0.036, 0.047, and 0.007, respectively) and yielded higher area under receiver operating characteristic values for predicting impaired scores than single-marker models based on plasma or cerebrospinal fluid viral load or CD4 count (area under receiver operating characteristic values = 0.71, 0.81, and 0.71, respectively) and in 4-marker models based on plasma sCD14 and 3 conventional markers compared with the 3-marker models. CONCLUSIONS: Plasma sCD14 is a biomarker associated with impaired neurocognitive testing in attention and learning domains in HIV infected individuals with advanced disease, suggesting involvement of cortical and limbic pathways by inflammatory processes in the cART era. Plasma sCD14 is a potential biomarker to monitor HAND progression and therapeutic responses. PMID- 21646913 TI - Chronic kidney disease and estimates of kidney function in HIV infection: a cross sectional study in the multicenter AIDS cohort study. AB - BACKGROUND: Cystatin C has been proposed as an alternative marker of kidney function among HIV-infected persons in whom serum creatinine is affected by extrarenal factors. METHODS: In this cross-sectional study, we compared estimated glomerular filtration rates (eGFR) using serum creatinine versus cystatin C between 150 HIV-uninfected and 783 HIV-infected men. We evaluated the prevalence of chronic kidney disease (CKD; eGFR less than 60 mL/min/1.73 m) and examined the influence of extrarenal factors on GFR estimates among HIV-infected men. RESULTS: Estimated GFRSCR was similar by HIV serostatus, but eGFRCYSC was lower in HIV infected men. A higher proportion of HIV-infected men were classified as having CKD when using eGFRCYSC versus eGFRSCR (7% vs 5%, P < 0.01). In HIV-infected individuals without CKD, eGFRSCR was higher than eGFRCYSC, whereas it was lower than eGFRCYSC in persons with CKD. In HIV-infected men, older age, proteinuria, and prior clinical AIDS were inversely associated with both GFR estimates. Higher serum albumin levels and angiotensin-converting enzyme inhibitor/angiotensin receptor blocker use were associated with lower eGFRSCR. HIV viral load, hepatitis C coinfection, and serum alkaline phosphatase were inversely associated with eGFRCYSC. CONCLUSION: Among HIV-uninfected and HIV-infected men of similar social risk behaviors, GFR estimates differed by biomarker and kidney function level. Estimated GFRCYSC classified a larger proportion of HIV-infected men with CKD compared with eGFRSCR. Differences between these GFR-estimating methods may be the result of the effects of extrarenal factors on serum creatinine and cystatin C. Until GFR-estimating equations are validated among HIV-infected individuals, current GFR estimates based on these biomarkers should be interpreted with care in this patient population. PMID- 21646914 TI - Reconstruction of the burnt perionychium: literature review and treatment algorithm. AB - Nail apparatus deformities associated with burn injuries can cause significant functional and aesthetic concerns. The majority of perionychial deformities result from eponychial retraction and scarring, whereas a minority are due to direct damage to the regenerative part of the nail complex. A variety of different techniques have been described for reconstruction including the use of local flaps, skin/composite grafts, and microvascular transfer. The authors review the different strategies available in the current burns literature and present an algorithm for the management of perionychial burn deformities. PMID- 21646915 TI - Simultaneous diagnosis of acute lymphoblastic leukemia and peripheral neuroblastic tumor in a child. AB - We report the case of a 3-year-old girl with a mediastinal mass, severe anemia, leukocytosis and neutropenia, in whom, after initial suspicion of metastatic neuroblastoma, a final diagnosis of concurrent ganglioneuroblastoma and acute lymphoblastic leukemia was made. The mediastinal tumor was surgically excised and the child subsequently underwent chemotherapy for acute lymphoblastic leukemia. The patient remains in complete remission from both diseases 4 years after the diagnosis and 24 months after completion of all treatment. The simultaneous occurrence of 2 different neoplasms in a child is very infrequent, and no comparable cases are reported in the literature. PMID- 21646916 TI - A home-based maintenance therapy program for acute lymphoblastic leukemia practical and safe? AB - The maintenance phase of treatment for childhood acute lymphoblastic leukemia is characterized by daily oral chemotherapy dose-adjusted on the basis of toxicity, monitored by regular (1 to 2 weekly) blood counts. A traditional approach is undertaking this at out-patient clinics. A home maintenance program was commenced to reduce visits to hospital and associated family disruption. The program organizes blood tests arranged to be taken at or near the patients' home. The results are examined by a pharmacist and specialist nurse; changes in therapy are communicated by telephone call and written confirmation. Hospital attendance is reduced to monthly visits. To assess the program, tablet counting and before-and after audits of parental satisfaction were undertaken. Results of the first 2 years are presented. Preliminary analysis to identify predictors of nonadherence was performed. Fifty families were included in the evaluation. There were no critical incidents. Poor adherence rates in the initial 3-month period (overall 24%) improved after increased support and advice were offered to 78%. Increasing age was correlated with good adherence (r=0.37, P=0.02). Partnership status of the child's caretakers was strongly associated with adherence (14% of poor adhering patients had caretakers in stable partnerships, compared with 87% of good adhering patients, P<0.01). PMID- 21646917 TI - Long-term neurocognitive outcomes in young adult survivors of childhood acute lymphoblastic leukemia. AB - Five-year survival rates of childhood acute lymphoblastic leukemia (ALL) exceed 80% due to central nervous system-directed treatment including cranial radiation (CRT) and chemotherapy. However, these treatments are associated with neurocognitive compromise, the extent of which is correlated with higher dose and younger age at treatment. The aims of this study were to explore long-term neurocognitive outcomes in adult survivors of childhood ALL, and to identify measures sensitive to neurotoxicity in long-term survivors. We examined 24 adults who received 18 Gy CRT and chemotherapy for treatment of ALL between ages 2 and 15 years (median, 5.5). Time since diagnosis ranged from 6 to 26 years (median, 16.6). Younger age at diagnosis and longer time since diagnosis were associated with lower scores on a computerized battery that requires speed and accuracy across a number of domains (MicroCog), and other standardized neurocognitive tests. When compared with population norms, MicroCog indices were below average in survivors diagnosed with ALL before age 5, but only the reasoning/calculation index was below average in survivors diagnosed with ALL after age 5. In contrast, intelligence quotient (IQ) scores were average. In addition to confirming earlier studies showing that younger children are more vulnerable to treatment-related neurotoxicity, here we show that deficits exist many years post treatment even with a relatively lower dose of CRT, and that these deficits are especially evident on tasks involving rapid processing of information. PMID- 21646918 TI - Survival to leukemia and lymphomas in children at a University Hospital in Cali, Colombia. AB - INTRODUCTION: Leukemia and lymphomas are the 2 most frequent malignancies among children in Cali, Colombia, although survival information for these malignancies remains limited in both Cali and throughout all of Colombia. OBJECTIVE: To estimate the 5-year survival rate in children diagnosed with leukemia and lymphomas in the University Hospital of Valle, at Cali, Colombia. METHODS: Three hundred and twenty-four patients younger than 15 years of age were included, diagnosed with either leukemia or lymphomas from 1998 to 2006. Active follow-up was conducted by clinical records reviewing and phone contacting. The cumulative survival rate was estimated to be from 1 to 5 years from diagnosis using the actuarial method. The Cox regression model was used to determine some of the factors associated with the prognosis. RESULTS: Of all cases, 61.8% were male patients, 75.3% corresponded to leukemia, and the rest to lymphomas. The global cumulated survival rate at 1 and 5 years were 71% and 50%, respectively. The risk of death from lymphomas was lower compared with leukemia, hazard ratio=0.36 (P<0.01). The highest cumulated survival rate was found in the group 5 to 9 years old (61%), followed by the group 0 to 4 years old (48%) and the group 10 to 14 years old (41%), all results reaching statistical significance (P<0.01), and showing lower survival rates than the data from international literature. CONCLUSIONS: The survival rates for leukemia and lymphomas in Cali are lower compared with the data of developed countries, suggesting that there are deficiencies in early diagnosis and in access to medication and opportune treatment. PMID- 21646919 TI - Severe allergic reactions to thiol-based cytoprotective agents mesna and amifostine in a child with a supratentorial primitive neuroectodermal tumor. AB - Both 2-mercaptoethane sulfonate sodium (mesna) and amifostine's active metabolite WR-1065 are thiol-based cytoprotective agents that are critical components of high-dose chemotherapy regimens used to treat various cancers in both adults and children. This case report describes a patient with a supratentorial primitive neuroectodermal tumor who developed severe drug reactions to both mesna and amifostine/WR-1065, suggesting that the thiol component of these agents triggered the adverse reactions. This report highlights the clinical presentation of drug induced hypersensitivity syndrome in the context of pediatric oncology and the supportive care measures that, if implemented rapidly, may diminish the reaction severity and allow successful completion of chemotherapy. PMID- 21646920 TI - Mitochondrial D-loop polymorphisms and mitochondrial DNA content in childhood acute lymphoblastic leukemia. AB - The mitochondrial displacement loop (D-loop) controls mitochondrial expression, with mutations and mitochondrial DNA (mtDNA) content linked to oncogenesis. We investigated D-loop polymorphisms and mtDNA content in childhood acute lymphoblastic leukemia (ALL). The D-loop was sequenced in 251 children: precursor B ALL (n=114), with 76 paired remission/relapse samples; T-ALL (n=24); cord blood controls (n=113). The mtDNA copy number was analyzed using real-time quantitative polymerase chain reaction for 92 controls and 54 ALL patients at diagnosis and remission. Polymorphisms around H-strand replication origin (nucleotides 150 to 199) and conserved sequence block II (nucleotides 299 to 317) were associated with leukemia biology and treatment response. T-ALL patients were more likely to have longer nt303 poly-C tract. T199C polymorphism was associated with increased risk of ALL in Malays; T152C was more frequent in good responders. There was no difference in mtDNA content between diagnostic ALL samples and controls; however, there was significant decrease in mtDNA content after treatment, especially in samples with OH polymorphisms. Somatic mutations were found in 13% (9 of 76) of patients, suggesting a link to leukemogenesis. Our results suggest that polymorphisms impacting transcriptional control could affect mtDNA replication. Decrease in mtDNA content after treatment may confer susceptibility to chemotherapy and be a clue to the good prognosis of childhood ALL. PMID- 21646921 TI - Adiposity, estrogen, and voice: the opera has just begun. PMID- 21646922 TI - From menarche to menopause: the fertile life span of celiac women. AB - OBJECTIVE: We evaluated menopause-associated disorders and fertile life span in women with celiac disease (CD) under untreated conditions and after long-term treatment with a gluten-free diet. METHODS: The participants were 33 women with CD after menopause (untreated CD group), 25 celiac women consuming a gluten-free diet at least 10 years before menopause (treated CD group), and 45 healthy volunteers (control group). The Menopause Rating Scale questionnaire was used to gather information on menopause-associated disorders. The International Physical Activity Questionnaire was used to acquire information on physical activity. RESULTS: Untreated celiac women had a shorter duration of fertile life span than did the control women because of an older age of menarche and a younger age of menopause (P < 0.01). The scores for hot flushes, muscle/joint problems, and irritability were higher in untreated celiac women than in the control women (higher by 49.4%, 121.4%, and 58.6%, respectively; P < 0.05). In comparison with untreated CD, long-lasting treatment of CD was not associated with a significant difference in the duration of fertile life span, but was only associated with a significant reduction in muscle/joint problems (a reduction of 47.1%; P < 0.05). CONCLUSIONS: Late menarche and early menopause causes a shorter fertile period in untreated celiac women compared with control women. A gluten-free diet that started at least 10 years before menopause prolongs the fertile life span of celiac women. The perception of intensity of hot flushes and irritability is more severe in untreated celiac women than in controls. Low physical exercise and/or poorer quality of life frequently reported by untreated celiac women might be the cause of reduced discomfort tolerance, thus increasing the subjective perception of menopausal symptoms. PMID- 21646923 TI - A prospective analysis of the association between cardiovascular disease and depression in middle-aged women. AB - OBJECTIVE: Experimental and clinical data demonstrate a close association between depression and coronary heart disease (CHD). Because no simple depression instrument for use by general practitioners has been shown to predict CHD, the objective of this study was to evaluate whether such a questionnaire could predict CHD. METHODS: The prevalence of CHD and CHD risk factors was assessed in women with depression, measured by a validated three-question depression screening instrument. Among 1,919 participants in the breast arterial calcification and CHD 5-year prospective study, 1,454 women (75.8%) completed a baseline depression inventory. RESULTS: Among the 1,454 women, 72.2% were postmenopausal, and the mean (SD) age at the conclusion of a 5-year prospective study was 61.3 (12.1) years. Among the women with no CHD risk factors at baseline and with one or less positive depression responses compared with those with two or more, 1.6% versus 3.8%, respectively, had at least one CHD risk factor (P = 0.004) by the 5th year of the study. In addition, 2.3% versus 6.0%, respectively, developed CHD (P = 0.005) by the 5th year of the study. Among all women with no positive depression responses compared with those with any positive depression responses, 2.1% versus 5.6%, respectively, had developed CHD by the 5th year of the study (P = 0.002). Finally, more positive depression responses were associated with a greater prevalence of CHD. CONCLUSIONS: In this prospective study, depression-detected by a validated three-item questionnaire-was able to predict those women more likely to develop CHD. PMID- 21646924 TI - Gene expression signatures differ with extent of atherosclerosis in monkey iliac artery. AB - OBJECTIVE: The aim of this study was to evaluate global gene expression patterns in the common iliac arteries of monkeys with a varied extent of atherosclerosis. METHODS: The left common iliac artery was removed from ovariectomized cynomolgus monkeys (n = 12) after 6.5 years of consuming a diet containing fat and cholesterol at levels comparable with those consumed in Western populations. Arterial gene expression was analyzed using DNA microarray and real-time reverse transcription-polymerase chain reaction. RESULTS: Significant differential expression of 986 genes was observed in iliac arteries containing moderate to large atherosclerotic plaques compared with normal/minimally affected reference group arteries. Atherosclerosis-associated genes included cytokines, chemokines, components of signal transduction pathways, and transcriptional activators and repressors, as well as other functional categories. Real-time reverse transcription-polymerase chain reaction confirmed a differential expression of genes chosen from a variety of functional categories. Specifically, the expression of genes for estrogen receptor-1, claudin 11, and brain heart protocadherin 7 was reduced, whereas the expression of genes for apolipoprotein E, growth differentiation factor 15, superoxide dismutase-2, SET domain bifurcated 2, phospholipase A2 group IIA, phospholipase A2 group VII, and ring finger protein 149 was increased in atherosclerotic arteries. CONCLUSIONS: The gene expression environment in arteries containing atherosclerotic plaques is profoundly different from that of relatively unaffected arteries and reflects the cellular and molecular complexity of atherosclerosis and associated arterial remodeling processes. PMID- 21646925 TI - Physical activity and sex hormone levels in estradiol- and placebo-treated postmenopausal women. AB - OBJECTIVE: Postmenopausal changes in the hormonal milieu in women with or without hormone therapy are hypothesized to be the pathway for a number of menopause associated modifications in physiology and disease risk. Physical activity may modify these changes in women's hormone profiles. The crucial yet complex relationship between physical activity and physiologic and pharmacologic sex hormone levels in postmenopausal women has not been investigated sufficiently. METHODS: Using structured recall, physical activity was assessed longitudinally during a period of 2 years in 194 postmenopausal women (90 randomized to 1 mg 17beta-estradiol treatment daily and 104 randomized to placebo) in the Estrogen in the Prevention of Atherosclerosis Trial. The levels of physical activity were correlated with the serum sex hormone and the serum hormone-binding globulin levels in each treatment group. RESULTS: Among the placebo-treated women, total energy expenditure was positively associated with sex hormone-binding globulin (SHBG; P < 0.001) and inversely associated with testosterones (total, bioavailable, or free) and androstenedione (P < 0.001 for all), as well as with estradiol (P = 0.02). In estradiol-treated women, estradiol levels were inversely associated with total energy expenditure (P = 0.002) and weekly hours spent in moderate or more vigorous physical activity (P = 0.001). CONCLUSIONS: Physical activity is associated with lower serum levels of estradiol in both hormone therapy-treated and untreated women. In placebo-treated women only, physical activity is associated with reduced androgen levels and elevated SHBG levels. PMID- 21646926 TI - Interactions of osteoporosis candidate genes for age at menarche, age at natural menopause, and maximal height in Han Chinese women. AB - OBJECTIVE: Age at menarche (AAM), age at natural menopause (ANM), and maximal height are closely related to bone mineral densities and osteoporosis. It is still unclear whether osteoporosis susceptibility genes are also associated with AAM, ANM, and maximal height in Chinese women. METHODS: In this relatively large cross-sectional sample of 722 Han Chinese postmenopausal women, 22 single nucleotide polymorphisms (SNPs) within 12 osteoporosis candidate genes that were identified from genome-wide association studies and replicated in our previous study were studied. The effects of a single gene on the AAM, ANM, and maximal height were investigated by linear regression analysis, whereas the gene-gene interactions were determined by a generalized multifactor dimensionality reduction method. RESULTS: It was revealed that the major histocompatibility complex (MHC) gene (rs3130340) was associated with ANM even after Bonferroni correction (P = 0.001). A significant gene-gene interaction for ANM involving rs3130340 in MHC, rs1038304 and rs4870044 in estrogen receptor-alpha gene (ESR1), and a significant three-SNP interaction model (SNP rs2273061 in jagged1, SNP rs6929137 in ESR1, and SNP rs2306033 in low-density lipoprotein receptor-related protein 4) for maximal height were identified. No single or combined effect of tested SNPs on AAM was discovered. CONCLUSIONS: Our study indicates that osteoporosis susceptibility SNPs, such as ESR1 (rs1038304, rs4870044, rs6929137), MHC (rs3130340), low-density lipoprotein receptor-related protein 4 (rs2306033), and jagged1 (rs2273061), might independently and/or in an interactive manner influence ANM and maximal height. All the SNPs tested had no association with AAM. PMID- 21646927 TI - Selective serotonin reuptake inhibitors and risk for major congenital anomalies. AB - OBJECTIVE: To estimate the risk of major congenital anomalies after exposure to selective serotonin reuptake inhibitors during pregnancy. METHODS: A retrospective cohort study based on national population-based registers (years 1996-2006) of births, congenital anomalies, and terminations of pregnancy because of severe fetal anomalies (maintained by National Institute for Health and Welfare, source offspring population n=635,583) and drug reimbursements (Social Insurance Institution) linked by a personal identification number. Offspring exposed to selective serotonin reuptake inhibitors during the first trimester (n=6,976) were compared with unexposed referent offspring. RESULTS: Overall major congenital anomalies were not more common in selective serotonin reuptake inhibitor-exposed offspring compared with unexposed referent offspring (adjusted odds ratio [OR] 1.08, 95% confidence interval [CI] 0.96-1.22). Fluoxetine was associated with an increased risk of isolated ventricular septal defects (adjusted OR 2.03, 95% CI 1.28-3.21) and paroxetine was associated with an increased risk of right ventricular outflow tract defects (adjusted OR 4.68, 95% CI 1.48-14.74). Citalopram use was associated with neural tube defects (adjusted OR 2.46, 95% CI 1.20-5.07). Fetal alcohol spectrum disorders were 10-times more common in the selective serotonin reuptake inhibitor-exposed offspring than in unexposed referent offspring. CONCLUSION: Fluoxetine use is associated with an increased risk of isolated ventricular septal defects and paroxetine is associated with right ventricular outflow tract defects. The absolute risk for these specific cardiac anomalies is small but should guide clinicians not to consider fluoxetine or paroxetine the first option when prescribing selective serotonin reuptake inhibitors to women planning pregnancy. Special attention should be given to alcohol use in pregnant women using selective serotonin reuptake inhibitors. PMID- 21646928 TI - Indications contributing to the increasing cesarean delivery rate. AB - OBJECTIVE: To examine physician-documented indications for cesarean delivery in order to investigate the specific factors contributing to the increasing cesarean delivery rate. METHODS: We analyzed rates of primary and repeat cesarean delivery, including indications for the procedure, among 32,443 live births at a major academic hospital between 2003 and 2009. Time trends for each indication were modeled to estimate the absolute and cumulative annualized relative risk of cesarean by indication over time and the relative contribution of each indication to the overall increase in primary cesarean delivery rate. RESULTS: The cesarean delivery rate increased from 26% to 36.5% between 2003 and 2009; 50.0% of the increase was attributable to an increase in primary cesarean delivery. Among the documented indications, nonreassuring fetal status, arrest of dilation, multiple gestation, preeclampsia, suspected macrosomia, and maternal request increased over time, whereas arrest of descent, malpresentation, maternal-fetal indications, and other obstetric indications (eg, cord prolapse, placenta previa) did not increase. The relative contributions of each indication to the total increase in primary cesarean rate were: nonreassuring fetal status (32%), labor arrest disorders (18%), multiple gestation (16%), suspected macrosomia (10%), preeclampsia (10%), maternal request (8%), maternal-fetal conditions (5%), and other obstetric conditions (1%). CONCLUSION: Primary cesarean births accounted for 50% of the increasing cesarean rate. Among primary cesarean deliveries, more subjective indications (nonreassuring fetal status and arrest of dilation) contributed larger proportions than more objective indications (malpresentation, maternal-fetal, and obstetric conditions). PMID- 21646929 TI - Intratympanic steroid injections as a salvage treatment for sudden sensorineural hearing loss: a randomized, double-blind, placebo-controlled study. AB - OBJECTIVE: The purpose of this study was to determine, through a randomized, double-blind, placebo-controlled trial, whether intratympanic steroid injections (ITSIs) could improve hearing recovery in patients with sudden sensorineural hearing loss (SSHL) who did not respond to initial systemic steroid therapy. STUDY DESIGN: This was a prospective, randomized, double-blind, placebo controlled study. SETTING: The study was conducted in 2 tertiary referral centers. PATIENTS: A total of 60 patients with idiopathic SSHL who did not respond to an initial round of systemic steroid therapy were included in this study. The subjects were randomized into an ITSI group and an intratympanic normal saline injection (ITNI) group, which were matched by age and sex. A total of 55 subjects completed the study. INTERVENTION: Participants received either ITSIs or ITNIs. Both groups received 4 injections within a 2-week period. MAIN OUTCOME MEASURES: Pure-tone thresholds were compared between the 2 groups 1 month after injection therapy. RESULTS: In the ITNI group, the pure-tone threshold was 69.9 +/- 18.5 dB before intratympanic injection therapy. After therapy, the hearing threshold improved by an average of 4.5 +/- 6.5 dB, and 10.7% of subjects improved by 10 dB or more. In the ITSI group, the pure-tone threshold was 64.6 +/ 17.7 dB before intratympanic injection therapy. After the therapy, the hearing threshold improved by an average of 9.8 +/- 8.5 dB, and 44.4% of subjects improved by 10 dB or more. Both the response rate and the level of hearing improvement were significantly greater in the ITSI group than in the ITNI group. CONCLUSION: These results demonstrate that ITSIs are beneficial as a salvage therapy for the treatment of patients with idiopathic SSHL who fail to respond to initial systemic steroid therapy. PMID- 21646930 TI - Neurotrophic receptors as potential therapy targets in postnatal development, in adult, and in hearing loss-affected inner ear. AB - HYPOTHESIS: The aim of this investigation was to define the expression of neurotrophic receptors within the developing inner ear of different postnatal ages. BACKGROUND: Pattern of differential expression of neurotrophic receptors provide molecular target sites for multifunctional nanoparticle-based cell specific therapeutics delivery to treat hearing diseases. METHODS: Protein expression of neurotrophic receptors was studied by immune-histochemistry, quantitative polymerase chain reaction, in situ hybridization, Western blot, in early and late postnatal, adult, and aging mice. RESULTS: There was a high correlation between results obtained at ribonucleic acid and protein levels. TrkB and TrkC gene expression increased during the first 2 weeks and also after the onset of hearing in adult mice. At the onset of hearing, TrkB-immunopositive staining occurred in inner hair cells and in cell bodies of spiral ganglion neurons. TrkC was detected in nerve endings beneath inner and outer hair cells and in supporting cells. Root cells within the spiral ligament and spiral ganglion neurons in the Rosenthal's canal showed high level of TrkC expression. p75NTR was found in organ of Corti similar to TrkC, and scattered neurons showed strong immunoreactivity in the Rosenthal's canal. PD540 mice, a model of age related hearing loss, showed a complete spiral ganglion cell loss in the basal turn. Although TrkB and TrkC were completely lacking in this region of the Rosenthal's canal, remaining nerve fibers were p75NTR immunopositive. CONCLUSION: We found differential expression pattern of TrkB, TrkC, and p75NTR receptors in the inner ear and could make a receptor expression data base. These findings, in turn, will help to design a study on receptor-specific drug targeting of the mice model of auditory development and aging. PMID- 21646931 TI - Clinical assessment of pitch perception. AB - OBJECTIVE: The perception of pitch has recently gained attention. At present, clinical audiologic tests to assess this are hardly available. This article reports on the development of a clinical test using harmonic intonation (HI) and disharmonic intonation (DI). STUDY DESIGN: Prospective collection of normative data and pilot study in hearing-impaired subjects. SETTING: Tertiary referral center. PATIENTS: Normative data were collected from 90 normal-hearing subjects recruited from 3 different language backgrounds. The pilot study was conducted on 18 hearing-impaired individuals who were selected into 3 pathologic groups: high frequency hearing loss (HF), low-frequency hearing loss (LF), and cochlear implant users (CI). INTERVENTION(S): Normative data collection and exploratory diagnostics by means of the newly constructed HI/DI tests using intonation patterns to find the just noticeable difference (JND) for pitch discrimination in low-frequency harmonic complex sounds presented in a same-different task. MAIN OUTCOME MEASURE(S): JND for pitch discrimination using HI/DI tests in the hearing population and pathologic groups. RESULTS: Normative data are presented in 5 parameter statistics and box-and-whisker plots showing median JNDs of 2 (HI) and 3 Hz (DI). The results on both tests are statistically abnormal in LF and CI subjects, whereas they are not significantly abnormal in the HF group. CONCLUSION: The HI and DI tests allow the clinical assessment of low-frequency pitch perception. The data obtained in this study define the normal zone for both tests. Preliminary results indicate possible abnormal TFS perception in some hearing-impaired subjects. PMID- 21646932 TI - Stapedectomy in sheep: an animal model for surgical training. AB - HYPOTHESIS AND BACKGROUND: Stapedectomy is a surgical technique that requires progressive training. The external and middle ear of sheep have a close resemblance to the human and have been previously used as surgical training models. In our project we describe the anatomy of the middle and external ear in sheep focusing on surgical landmarks and technique in order to determine whether the sheep's ear is an adequate model for stapedectomy training. MATERIALS AND METHODS: We reviewed the literature on sheep anatomy and use of sheep as an otologic surgical model. Macroscopic sections as well as temporal bone computed tomography were obtained. Stapedectomy was performed on 40 sheep, using 4 mm platinum piston prosthesis, by first year residents. RESULTS: Most of the structures in the sheep's middle ear are similar to those in humans although their size is about two thirds smaller. Incus long process is shorter, thicker, and closer to the malleus body, making the piston insertion and adjustment more difficult. The median surgical time of stapedectomy was reduced from 70 (52.5 100.3) minutes to 39.5 (35.5-48) minutes after completing training. There was also a reduction in rate of complications (flap disruption, incus dislocation, and footplate mobilization). CONCLUSION: Sheep ears constitute a cheap, easy to obtain and anatomically adequate model for stapedectomy training. PMID- 21646933 TI - Intratympanic dexamethasone/hyaluronic acid mix as an adjunct to intravenous steroid and vasoactive treatment in patients with severe idiopathic sudden sensorineural hearing loss. AB - OBJECTIVE: To evaluate differences in effectiveness (hearing recovery rates) between idiopathic sudden sensorineural hearing loss (ISSNHL) patients treated with intravenous therapy alone and patients treated with a combination of intravenous and intratympanic therapy. STUDY DESIGN: Retrospective case review. SETTING: Tertiary referral hospital center. PATIENTS AND INTERVENTIONS: Ninety four patients with moderate ISSNHL treated with an intravenous steroid and vasoactive regimen (duration of therapy, 9 +/- 2.76 d) and 76 patients with severe ISSNHL treated with a combination regimen of intravenous and intratympanic therapy (duration of therapy, 10 +/- 2.71 d) were reviewed. In the latter patients' group, a series of 3 intratympanic injections of a dexamethasone/hyaluronic acid mix solution were applied every 2 days. MAIN OUTCOME MEASURE: Pure-tone audiometric thresholds at 0.5, 1, 2, 4, and 8 kHz were compared between groups using the Wilcoxon test. RESULTS: Combination therapy in severe ISSNHL did not show any statistically significant difference in effectiveness to intravenous therapy in moderate ISSNHL (p > 0.05). CONCLUSION: In patients with severe ISSNHL, starting intratympanic steroid therapy as an adjunct early in the course of intravenous steroid and vasoactive therapy improves hearing to a level which is obtained in patients with less severe (moderate) ISSNHL treated with intravenous therapy alone. PMID- 21646934 TI - Comparisons of sound processors based on osseointegrated implants in patients with conductive or mixed hearing loss. AB - OBJECTIVES: To compare the patient benefit of the Bone-Anchored Hearing Aid (Baha) Divino and the Baha BP100 sound processors. STUDY DESIGN: Prospective experimental study. SETTING: Tertiary referral center. PATIENTS: Twenty experienced Baha Divino users with conductive or mixed hearing loss. INTERVENTIONS: Speech understanding in quiet and in noise was measured in unaided conditions and with the subjects' own Divino sound processors. Speech in noise was tested with the noise arriving either from the front or from the back of the subjects. Then, participants were fitted with BP100 sound processors. After 3 months of use, all speech tests were repeated with the new sound processor along with a subset of the tests from the first session to assess learning or training effects. The Abbreviated Profile of Hearing Aid Benefit (APHAB) questionnaires were administered at 0 and at 3 months. MAIN OUTCOME MEASURE: Aided speech understanding in quiet and in noise. RESULTS: No significant differences were found between the two devices regarding speech understanding in quiet. Speech understanding in noise was improved, on average, by +1.0 to +2.3 dB with the BP100 when compared to the Divino. No significant learning effects were found. Subjective ratings assessed by the Abbreviated Profile of Hearing Aid Benefit (APHAB) questionnaire showed statistically significantly better scores for the BP100 for noisy or reverberant environments, when compared with the Divino. CONCLUSION: The performance of both devices is similar for speech understanding in quiet. Speech understanding in noise is significantly better with the Baha BP100 than with the Baha Divino, presumably as a result of the improved signal processing. PMID- 21646935 TI - A randomized, controlled study comparing the effects of vestipitant or vestipitant and paroxetine combination in subjects with tinnitus. AB - OBJECTIVE: Tinnitus is a common symptom that demonstrates a significant comorbidity with anxiety and depression. The novel neurokinin-1 receptor antagonist, vestipitant, has anxiolytic properties and a good safety profile. Vestipitant was investigated for potential effect against chronic tinnitus as a stand-alone treatment and in conjunction with a selective serotonin reuptake inhibitor, paroxetine. STUDY DESIGN: Randomized, double-blind, crossover study. SETTING: Tertiary neurotologic and audiologic center with additional referrals from a secondary university hospital center. PATIENTS: Twenty-four adult patients with tinnitus were randomized into the study. MAIN OUTCOME MEASURES: Visual analogue scale (VAS) measurements of tinnitus loudness (intensity), pitch and distress, VAS measurements of arousal/anxiety, Tinnitus Handicap Inventory, Quick Inventory of Depressive Symptomatology, and plasma concentrations of trial drugs. RESULTS: No statistically significant treatment benefit effect was detected for tinnitus (intensity, pitch, and distress) VAS scores, arousal-anxiety VAS scores, Tinnitus Handicap Inventory, or tinnitus aggravation scores assessed on Days 1 and 14. However, a statistically significant worsening of tinnitus intensity and distress scores was observed after vestipitant compared with placebo for the mean data collected over the treatment period. No relevant differences in vestipitant plasma concentrations were observed between the subjects given the combination with paroxetine and those receiving vestipitant alone. No specific relationships were observed between tinnitus intensity and vestipitant plasma concentrations. CONCLUSION: Although well-tolerated vestipitant, alone or in combination with paroxetine, was not effective in ameliorating tinnitus in this patient group. PMID- 21646936 TI - Cochlear implantation after bacterial meningitis in children: outcomes in ossified and nonossified cochleas. AB - OBJECTIVE: To investigate the outcome of cochlear implantation in children with postmeningitic deafness and to compare the outcomes in ossified and nonossified cochleas. STUDY DESIGN: A retrospective case note review of children with postmeningitic deafness requiring cochlear implantation. SETTING: Manchester Auditory Implant Centre--a tertiary referral center. PATIENTS: Children younger than 16 years with postmeningitic deafness. INTERVENTION: Cochlear implantation to aid auditory habilitation. MAIN OUTCOME MEASURE: Outcomes were measured using Categories of Auditory Performance Score and the Manchester Speech and Language Development Scale. RESULTS: Fifty-two children underwent cochlear implantation. Patients were allocated to 2 groups--Group 1 with nonossified cochleas and Group 2 with cochlear ossification. Group 1 comprised 25 children, of whom, 22 had Categories of Auditory Performance Score of 5 or higher. Nineteen of them were able to use simple phrases of 3 words or more on the Manchester Speech and Language Development Scale. Group 2 comprised 27 children. Fourteen had partial ossification with complete standard electrode insertion in all instances. Of the 13 children with gross ossification, 3 had scala vestibuli insertion, 7 had split electrode insertion and 3 had partial electrode insertion. In this group, 20 children had Categories of Auditory Performance Score of 5 or higher. Nineteen children were able to use simple phrases of 3 words or more on Manchester Speech and Language Development Scale. CONCLUSION: Children with postmeningitic deafness benefit significantly from cochlear implantation. However, the audiologic outcomes are difficult to predict, especially in the presence of cochlear ossification. Acquisition of speech and language in these children after cochlear implantation is possible even in the presence of gross cochlear ossification. PMID- 21646937 TI - Soft tissue complications after pediatric cochlear implantation in children younger than 12 months. AB - OBJECTIVES: Children are undergoing cochlear implantation younger than ever before. There has been some concern that young children may have an increased risk of soft tissue complications than older age groups. We aim to review the major and minor soft tissue complications after pediatric cochlear implantation in the age group of younger than 12 months. STUDY DESIGN: Retrospective case review. METHODS: Patients were identified from the cochlear implant program database of more than 1,000 children at the Hospital for Sick Children, Toronto, Canada. Demographic data, cause of hearing loss, and time of the onset of hearing loss were recorded. RESULTS: A total of 66 patients were identified (94 implants) in the age group of younger than 12 months. Of these, there was 1 minor complication (implanted at 8 mo)-skin infection around implant 14 days later treated with antibiotics. There were no major complications. CONCLUSION: Cochlear implantation in children younger than 12 months does not increase the risk of soft tissue complications. PMID- 21646938 TI - Rotavirus diarrhea among children less than 5 years of age in urban Ghana. AB - We collected clinical and morphologic data from children with diarrhea attending 3 diverse hospitals/clinics in Accra. Stool samples were tested for rotavirus and Cryptosporidium spp. In all, 58% of the children with diarrhea had rotavirus infections, 25% of which were of the G3 sero/genotype. The most common strains were G3P [6] (18.8%) and G2P [6] (12.5%). Cryptosporidium spp. infections were uncommon (3/143, 2.0%). PMID- 21646939 TI - Intrathoracic pressure regulation improves 24-hour survival in a pediatric porcine model of hemorrhagic shock. AB - Hemorrhagic shock is a common cause of mortality and morbidity in the pediatric population. Intrathoracic pressure regulation (IPR) lowers intrathoracic pressure, thereby decreasing intracranial pressure and increasing venous return, cardiac output, and cerebral perfusion without the need for immediate fluid resuscitation. We hypothesized that IPR would improve hemodynamics and 24-h survival in a pediatric porcine model of hemorrhagic shock. Twenty piglets were subjected to a 50% total blood volume hemorrhage over 15 min and then randomized to treatment with either IPR or no treatment. After 60 min, survivors were autotransfused, weaned from the ventilator, and assessed and autopsied at 24 h. Mean arterial pressures (MAPs), cardiac index (CI), and arterial blood gases were recorded. MAP (mm Hg) was significantly higher in the IPR group (60.8 +/- 3.7) versus controls (41.2 +/- 4.6, p < 0.01). Mean CI (L/min/m2) was significantly higher with IPR (3.9 +/- 0.24) versus controls (2.5 +/- 0.39, p < 0.01). IPR survival rates were significantly improved with IPR [9/9 (IPR) versus 5/11 (controls); p < 0.02]. In this piglet model of hemorrhagic shock, IPR treatment safely and significantly improved MAP, CI, and 24-h survival rates. PMID- 21646940 TI - Baseline values of candidate urine acute kidney injury biomarkers vary by gestational age in premature infants. AB - Acute kidney injury (AKI) is common in premature infants and is associated with poor outcomes. Novel biomarkers can detect AKI promptly. Because premature infants are born with underdeveloped kidneys, baseline biomarker values may differ. We describe baseline values of urinary neutrophil gelatinase-associated lipocalin (NGAL), IL-18, kidney injury molecule-1 (KIM-1), osteopontin (OPN), beta-2 microglobulin (B2mG), and Cystatin-C (Cys-C). Next, we test the hypothesis that these biomarkers are inversely related to GA. Candidate markers were compared according to GA categories in 123 infants. Mixed linear regression models were performed to determine the independent association between demographics/interventions and baseline biomarker values. We found that urine NGAL, KIM-1, Cys-C, and B2mG decreased with increasing GA. With correction for urine creatinine (cr), these markers and OPN/cr decreased with increasing GA. IL 18 (with or without correction for urine creatinine) did not differ across GA categories. Controlling for other potential clinical and demographic confounders with regression analysis shows that NGAL/cr, OPN/cr, and B2mG/cr are independently associated with GA. We conclude that urine values of candidate AKI biomarkers are higher in the most premature infants. These findings should be considered when designing and analyzing biomarker studies in newborn with AKI. PMID- 21646941 TI - Systematic evaluation of exertional hyperthermia in children and adolescents with hypohidrotic ectodermal dysplasia: an observational study. AB - To evaluate exertional overheating and the impact of physical exercise on individuals with hypohidrotic ectodermal dysplasia (HED) and to assess protective effects of cooling devices, 13 boys and male adolescents with X-linked HED (XLHED) and age-matched healthy male controls were studied during standardized exercise on a bicycle ergometer at ambient temperatures of 25 and 30 degrees C, without cooling and with evaporative skin cooling devices at 30 degrees C. Body core temperature during and after exercise, heart rate, performance, endurance, and serum lactate were investigated. XLHED subjects experienced a significantly greater rise in body temperature after cycling than healthy controls, and their body temperature remained elevated longer. Maximum heart rates and lactate values did not differ significantly between XLHED and control groups. Application of skin cooling devices led to a clinically relevant attenuation of exertional hyperthermia in XLHED patients, and a previous tendency toward lower performance disappeared. This first systematic study of the effects of physical exercise on HED patients demonstrates a rapid and lasting body temperature increase in XLHED subjects after cycling, posing them at risk of exercise-induced hyperthermia. External evaporative skin cooling attenuates exertional overheating in HED patients and may facilitate their participation in athletic activities and professional life. PMID- 21646942 TI - Patterns of blood protein concentrations of ELGANs classified by three patterns of respiratory disease in the first 2 postnatal weeks. AB - We examined the association between elevated concentrations of 25 blood proteins in blood spots collected on postnatal d 1, 7, and 14 from infants <28 wk gestation who survived to 24 mo and the risk of two patterns of early lung disease i.e. early and persistent pulmonary dysfunction (EPPD) and normal early pulmonary function followed by pulmonary deterioration (PD). Thirty-eight percent (n = 347) of our cohort had PD, and 43% (n = 383) had EPPD. On postnatal d 14, elevated concentrations of two proteins (RANTES and VEGF) were associated with reduced risk of PD. Similarly, the risk of EPPD was also reduced if three proteins had elevated concentrations on postnatal d 14 (RANTES, MMP-1, and VEGF). In contrast, the risk of EPPD was increased if on d 14 two proteins had elevated concentrations (IL-8 and ICAM-1). Inflammation might influence the risk of EPPD and PD or be a consequence of lung damage or therapies to minimize lung dysfunction. PMID- 21646943 TI - Insulin administration for treatment of pediatric diabetic ketoacidosis: are lower rates of infusion beneficial? PMID- 21646944 TI - The practice of mechanical ventilation in pediatrics: science, art, or a simple routine? PMID- 21646945 TI - A framework for healing. PMID- 21646946 TI - Renal function and extracorporeal membrane oxygenation: the crossroads of concurrent multiple organ support. PMID- 21646947 TI - Extracorporeal membrane oxygenation survivors and pulmonary function: encouraging outcomes early in life. PMID- 21646948 TI - Biomarkers for sepsis: PERSEVERE to the bitter end. PMID- 21646949 TI - Red blood cell transfusions in children: treat the child, not the number! PMID- 21646950 TI - It is what it was. PMID- 21646951 TI - Cortisol-binding globulin: more than just a carrier? PMID- 21646952 TI - Leaks cause problems not only in Washington politics! Has the time come for cuffed endotracheal tubes for newborn ventilation? PMID- 21646953 TI - Development of an accurate score to predict early-onset neonatal sepsis. PMID- 21646954 TI - Acute kidney injury treatment and the optimization of diuretics in newborns. PMID- 21646955 TI - Global versus local bioethics. PMID- 21646956 TI - Let the pediatric intensive care unit's door open: but not to all winds! PMID- 21646957 TI - Imaging in severe malaria. PMID- 21646958 TI - What does it take to get some peace and quiet in the pediatric intensive care unit? PMID- 21646959 TI - Early exchange and pheresis therapies in critical pertussis. PMID- 21646961 TI - Factors that influence stabilization times in children requiring transport. PMID- 21646962 TI - Prediction of serotypes causing invasive pneumococcal disease in unvaccinated and vaccinated populations. AB - INTRODUCTION: Before the introduction of the heptavalent pneumococcal conjugate vaccine (Prevnar-7), the relative prevalence of serotypes of Streptococcus pneumoniae was fairly stable worldwide. We sought to develop a statistical tool to predict the relative frequency of different serotypes among disease isolates in the pre- and post-Prevnar-7 eras using the limited amount of data that is widely available. METHODS: We initially used pre-Prevnar-7 carriage prevalence and estimates of invasiveness derived from case-fatality data as predictors for the relative abundance of serotypes causing invasive pneumococcal disease during the pre- and post-Prevnar-7 eras, using negative binomial regression. We fit the model to pre-Prevnar-7 invasive pneumococcal disease data from England and Wales and used these data to (1) evaluate the performance of the model using several datasets and (2) evaluate the utility of the country-specific carriage data. We then fit an alternative model that used polysaccharide structure, a correlate of prevalence that does not require country-specific information and could be useful in determining the postvaccine population structure, as a predictor. RESULTS: Predictions from the initial model fit data from several pediatric populations in the pre-Prevnar-7 era. After the introduction of Prevnar-7, the model still had a good negative predictive value, though substantial unexplained variation remained. The alternative model had a good negative predictive value but poor positive predictive value. Both models demonstrate that the pneumococcal population follows a somewhat predictable pattern even after vaccination. CONCLUSIONS: This approach provides a preliminary framework to evaluate the potential patterns and impact of serotypes causing invasive pneumococcal disease. PMID- 21646963 TI - Above all, it must be consulted. PMID- 21646964 TI - Teaching physician numeracy. PMID- 21646966 TI - We need a new model for obtaining students' consent to conduct peer physical examinations. PMID- 21646968 TI - Commentary: improving the supply and distribution of primary care physicians. AB - The current medical education system and reimbursement policies in the United States have contributed to a maldistribution of physicians by specialty and geography. The causes of this maldistribution include financial barriers that prevent the individuals who would be the most likely to serve in primary care and underserved areas from entering the profession, large taxpayer subsidies to teaching hospitals that provide incentives to act in ways that are not in the best interest of society, and reimbursement policies that discourage physicians from providing primary care. The authors propose that the maldistribution of physicians can be addressed successfully by reducing the financial barriers to becoming a primary care physician, aligning subsidies with societal interests, and providing financial incentives that target primary care. They suggest that the Patient Protection and Affordable Care Act of 2010 takes steps in the right direction but that more financially prudent measures should be taken as politicians revisit health care reform with heightened financial scrutiny. PMID- 21646969 TI - Commentary: charting a course for success: excellence in clinical care and discovery in academic departments. AB - The current shifts in academics not only invite new challenges but create previously unexplored opportunities for unique discoveries in health. Leaders in academic departments must consider changes in academic medicine as new courses to be charted rather than an inevitable shifting of the ground beneath them. Under this model, clinical excellence is coupled with discovery, where trainees, faculty, and patients and families are continually exposed to asking questions and identifying ways to move science forward to improve health. Academic pediatrics remains today a vibrant and exciting discipline with extraordinary leaders and committed trainees. We must continue to inspire on the voyage to excellence, keeping our eyes on the horizon and not the gathering storms. PMID- 21646970 TI - Commentary: concentrating the mind. AB - Among numerous threats to the academic mission, diminishing funding for biomedical research attracts particular attention because it is so tangible and is likely to slow scientific progress in the near term. More dangerous still may be long-term adverse effects of reduced resources on the pipeline of physician scientists. For pediatric departments, the impact may be accentuated by an already disproportionately low share of research funding and by well-recognized challenges in meeting parallel clinical demand for subspecialty services. In the absence of an imminent prospect of increased federal or foundational support, solutions to these workforce and economic issues will need to come from a reassessment of priorities within departments and institutions. Leaders in academic pediatrics must recognize the importance of all aspects of the academic mission but also recognize that within a single institution, investments must be tied to people and programs with a high likelihood of success. Proper alignment of investments and capabilities at both the individual and institutional levels will give academic health centers the chance to redefine themselves and succeed in a threatening environment. PMID- 21646971 TI - Commentary: professionalism, unionization, and physicians' strikes. AB - Professionalism is the basis of medicine's social contract with society. The details of that contract are influenced by the presence or absence of a national health plan. In countries with such a plan, unlike in the United States, negotiations are dictated by the nature of medicine's contract with society and take place between the medical profession and society directly. This system has required that medicine be represented at the negotiating table, and, in most instances, it has resulted in the unionization of physicians. To influence these negotiations, the medical profession has used various forms of collective action, including strikes. As the United States continues on the path toward health care reform, it seems likely that the American medical profession will also require an organization to represent it at the negotiating table and will be under the same pressures to strike as are physicians in other countries. Because both unionization and strikes pose potential threats to the professionalism of students, residents and practicing physicians, such issues should be a part of the medical education curriculum at both the undergraduate and postgraduate levels. The authors briefly review the literature on strikes and job actions and share personal experiences to support this discussion. Students and residents should have an opportunity to consider these issues in a safe environment, both to understand the potential impact of a strike on patients and the profession and to determine their own personal course of action should such a situation arise. PMID- 21646972 TI - Commentary: experience with resident unions at one institution and implications for the future of practicing physicians. AB - This commentary discusses the forces behind the formation of a resident union at the University of New Mexico School of Medicine and the union's evolution over its first three years. Because unions exist primarily to provide an avenue for advocacy to their members, they could have a negative impact on resident professionalism and on the faculty-resident mentor relationship. Resident unionization could also adversely impact the perceived balance between education and clinical service, to the detriment of the professional identity development of resident physicians. Despite this concern, the authors express their initial, cautious optimism that the union is instead currently promoting resident professionalism. The resident union has provided a forum for a unified resident voice, the engagement of the residents in safety and quality improvement activities, and advocacy for, and direction of, additional patient care funds, all of which has encouraged resident professionalism. Residents who have been active in the union also seem to have maintained altruistic professional attitudes as well as engagement in their educational activities. However, as the environment changes from one of increasing resources to one of stagnant or decreasing institutional resources, inevitable conflicts will arise between advocacy for resident salaries and benefits and patient care needs, and the manner in which the resident union will balance these conflicting needs and what impact it will have on the residents' professional identity development is unclear. PMID- 21646973 TI - What I know about excruciating physical pain. PMID- 21646974 TI - Artist's statement: fifteen words by a 13th-century philosopher Saint. PMID- 21646975 TI - Medicine and the arts. Big Bambu: by Doug Starn and Mike Starn. Commentary. PMID- 21646977 TI - AM last page. Patient-centered medical home. PMID- 21646978 TI - Ultraviolet absorption by contact lenses and the significance on the ocular anterior segment. AB - It is well established that both acute and chronic ultraviolet (UV) exposure can lead to various ophthalmic pathologic conditions in the anterior segment. Several scientific studies have demonstrated that after UV exposure, the unprotected cornea is vulnerable to damage in the epithelial, stromal, and endothelial cellular layers. DNA damage, apoptosis, and altered protease expression are all examples of harmful changes that can occur within the cornea after irradiation. Beyond the cornea, damage associated with UV exposure, such as decreased antioxidant levels and increased reactive oxygen species production, has been noted in the aqueous humor and crystalline lens. Ultraviolet-blocking contact lenses have the potential to provide protection against such exposure to harmful UV radiation. Experimental use of UV-absorbing contact lenses prevented detrimental cellular changes to the cornea and maintained corneal clarity after UV exposure. Additionally, studies suggest that shielding the aqueous humor and crystalline lens from irradiation with UV-absorbing contact lenses aids in protection against precataractous changes. Despite ongoing research, to date, neither chronic nor clinical studies have been performed in humans to demonstrate that wearing UV-blocking contact lenses reduces the risk of developing cataracts or other ocular disorders within the anterior segment. This article will discuss the impact of UV exposure on ocular tissue and the need for adequate UV protection, with particular emphasis on UV-blocking contact lenses. PMID- 21646979 TI - A review: role of ultraviolet radiation in age-related macular degeneration. AB - Age-related macular degeneration (AMD) is a leading cause of blindness in the western world. The retina is highly susceptible to photochemical damage from continuous exposure of light and oxygen. The cornea and the lens block a major portion of the ultraviolet (UV) radiation from reaching the retina (<295 nm). The relationship between UV light exposure and AMD is unclear, although short wavelength radiation and the blue light induce significant oxidative stress to the retinal pigment epithelium. Epidemiologic evidence indicates a trend toward association between severity of light exposure and AMD. In this review, we discuss type 1 and type 2 photochemical damage that occurs in response to UV exposure. We examine the impact of different doses of exposure to UV radiation and the subsequent production of oxidative stress in AMD. Local and systemic protective mechanisms of the retina including antioxidant enzymes and macular pigments are reviewed. This article provides a review of possible cellular and molecular effects of UV radiation exposure in AMD and potential therapies that may prevent blindness resulting from this disease. PMID- 21646980 TI - Ultraviolet phototoxicity to the retina. AB - OBJECTIVE: This overview of ultraviolet (UV) phototoxicity considers the interaction of UVA and short-wavelength VIS light with the retina and retinal pigment epithelium. METHODS: The damage mechanisms underlying UV retinal phototoxicity are illustrated with a literature survey and presentation of experimental results. RESULTS: Depending on the wavelength and exposure duration, light interacts with tissue by three general mechanisms: thermal, mechanical, or photochemical. Although the anterior structures of the eye absorb much of the UV component of the optical radiation spectrum, a portion of the UVA band (315-400 nm) penetrates into the retina. Natural sources, such as the sun, emit energetic UV photons in relatively long durations, which typically do not result in energy confinement in the retina, and thus do not produce thermal or mechanical damage but are capable of inducing photochemical damage. Photochemical damage in the retina proceeds through Type 1 (direct reactions involving proton or electron transfers) and Type 2 (reactions involving reactive oxygen species) mechanisms. Commonly used drugs, such as certain antibiotics, nonsteroidal anti-inflammatory drugs, psychotherapeutic agents, and even herbal medicines, may act as photosensitizers that promote retinal UV damage, if they are excited by UVA or visible light and have sufficient retinal penetration. CONCLUSIONS: Although the anterior portion of the eye is the most susceptible to UV damage, the retina is at risk to the longer UV wavelengths that propagate through the ocular media. Some phototoxicity may be counteracted or reduced by dietary intake of antioxidants and protective phytonutrients. PMID- 21646981 TI - The effect of triceps repair techniques following olecranon excision on elbow stability and extension strength: an in vitro biomechanical study. AB - OBJECTIVES: To determine the effect of two types of triceps repair techniques on elbow stability and extension strength in the setting of olecranon deficiency using a cadaveric model. METHODS: Eight fresh-frozen cadaveric arms were tested in an elbow motion simulator, which produced active elbow extension by applying physiological loads to the tendons. Computed tomography-based surface models were used to determine cutting planes corresponding to sequential levels of olecranon resection. Both anterior and posterior triceps repairs were simulated for each level. RESULTS: Progressive sectioning of the olecranon increased elbow laxity for both active and passive extension (P < 0.001). Although the posterior repair resulted in greater laxity than the anterior repair for all but the 50% resection, this difference was small (less than 3 degrees ) and not statistically significant for either active (P = 0.2) or passive (P = 0.1) extension. Active extension produced less joint laxity than passive extension for both the anterior (P = 0.007) and posterior (P = 0.001) repairs. The posterior repair provided greater extension strength than the anterior repair at all applied triceps tensions and for all olecranon resections (P = 0.01). Both repairs reduced extension strength relative to the intact state (P < 0.01). Sequential olecranon excision decreased extension strength (P = 0.04); however, there were no differences between resection levels (P > 0.05). CONCLUSIONS: On average, there was a loss of extension strength of 24% and 30% for the posterior and anterior repairs, respectively. There was no significant difference in stability between repair techniques. Posterior repair of the triceps after olecranon excision would thus appear to be efficacious as a result of its higher triceps extension strength. However, clinical studies are needed to confirm these in vitro observations. PMID- 21646982 TI - End-user evaluations of a personal computer-based pediatric advanced life support simulator. AB - INTRODUCTION: : To inform further development of a personal computer-based Pediatric Advanced Life Support (PALS) simulator, we wished to understand users' perceptions of this new technology. Specifically, we sought to determine whether the simulator was perceived as an effective training tool, whether it filled a gap in the users' current training regimen, and whether these perceptions were impacted by professional affiliation and PALS training history. METHODS: : We surveyed multidisciplinary health care workers in a tertiary care pediatric hospital who used our simulator. RESULTS: : A total of 789 users completed an evaluation of the simulator. Ninety-five percent of respondents agreed that the PALS simulator is an effective educational tool. Eighty-nine percent agreed that the simulator filled a gap in their training, although physicians agreed with this statement more strongly than nurses (P = 0.001). Prior resuscitation training history did not impact whether users perceived that the simulator filled a curricular gap. Users most commonly cited the simulator's realism, its capacity to facilitate practice, and its help feature as the top three qualities they most appreciated. Users' top three suggestions for improving the simulator included provision of a structured tutorial, specific user interface improvements, and encouragement of more widespread access to the simulator. CONCLUSIONS: : Hospital based pediatric providers are open to using personal computer-based simulation to provide on-demand refresher training in the cognitive aspects of PALS. Through its capacity to reach a large number of health care workers without the need for instructor presence, this technology could be used to help develop a more targeted role for mannequin simulation. PMID- 21646983 TI - Team communication with patient actors: findings from a multisite simulation study. AB - INTRODUCTION: Patient satisfaction is an important healthcare outcome and communication with clinical staff is an important determinant. Simulation could identify problems and inform corrective action to improve patient experience. METHODS: One hundred eight randomly selected maternity professionals in 18 teams were videoed managing a patient-actor with a simulated emergency. The trained patient-actor assessed the quality of staff-patient interaction. Clinicians scored teams for their teamwork skills and behaviors. RESULTS: There was significant variation in staff-patient interaction, with some teams not having exchanged a single word and others striving to interact with the patient-actor in the heat of the emergency. There was significant correlation between patient actor perceptions of communication, respect, and safety and individual and team behaviors: number, duration, and content of communication episodes, as well as generic teamwork skills and teamwork behaviors. The patient-actor perception of safety was better when the content of the communication episodes with them included certain items of information, but most teams failed to communicate these to the patient-actor. CONCLUSION: Some aspects of staff-patient interaction and teamwork during management of a simulated emergency varied significantly and were often inadequate in this study, indicating a need for better training of individuals and teams. PMID- 21646984 TI - Relationship between standardized patient checklist item accuracy and performing arts experience. AB - INTRODUCTION: : It is not known whether a Standardized Patient's (SP's) performing arts background could affect his or her accuracy in recording candidate performance on a high-stakes clinical skills examination, such as the Comprehensive Osteopathic Medical Licensing Examination Level 2 Performance Evaluation. The purpose of this study is to investigate the differences in recording accuracy of history and physical checklist items between SPs who identify themselves as performing artists and SPs with no performance arts experience. METHODS: : Forty SPs identified themselves as being performing artists or nonperforming artists. A sample of SP live examination ratings were compared with a second set of ratings obtained after video review (N = 1972 SP encounters) over 40 cases from the 2008-2009 testing cycle. Differences in SP checklist recording accuracy were tested as a function of performing arts experience. RESULTS: : Mean overall agreement rates, both uncorrected and corrected for chance agreement, were very high (0.94 and 0.79, respectively, at the overall examination level). There was no statistically significant difference between the two groups with respect to any of the mean accuracy measures: history taking (z = -0.422, P = 0.678), physical examination (z = -1.453, P = 0.072), and overall data gathering (z = -0.812, P = 0.417) checklist items. CONCLUSION: : Results suggest that SPs with or without a performing arts background complete history taking and physical examination checklist items with high levels of precision. Therefore, SPs with and without performing arts experience can be recruited for high-stakes SP-based clinical skills examinations without sacrificing examination integrity or scoring accuracy. PMID- 21646985 TI - Eco-friendly laparoscopic home trainer. AB - INTRODUCTION: : Laparoscopic surgery is becoming the main surgical technique in use today. Surgical trainees have to be able to practice these skills in a safe environment. This article describes the design of a novel cheap home laparoscopic trainer using recycled and reusable items. METHODS: : This novel home laparoscopic trainer is designed using a mobile phone, torch, and shoe box. Fifteen surgical trainees with variable laparoscopic experience used the device and provided feedback by filling in a Likert scale questionnaire. RESULTS: : This is a device that is easy to make and reuse with equipment that is easily accessible in most environments. All the trainees who used the device found it easy to use and helpful for practicing hand-eye coordination. CONCLUSIONS: : This is simple and low-cost device allows trainees to practice laparoscopic skills in a safe environment. It provides a design that is accessible and recyclable, hence useful as a low-technology device in places where finances are limited. PMID- 21646986 TI - Anaphylaxis simulation session: seeing is believing. AB - INTRODUCTION: : Massive anaphylactic reactions are rare but potentially life threatening events. It is recognized that there are concerns in both the immediate management and recognition of the condition. The report describes an innovative approach to develop a simple, low-cost device to simulate a rash on a mannequin as part of an education session. METHODS: : A system was developed to allow a plastic sheet with surface-mounted light-emitting diodes to be placed under the mannequin's chest skin. The light-emitting diodes presence and intensity could be controlled remotely via a potentiometer. RESULTS: : Staff were initially surprised by the presence of a rash on the mannequin but very quickly used the presence of the rash to support a diagnosis of severe anaphylaxis. Participants appeared to have responded positively to these simulation- reinforced courses on acute anaphylaxis. CONCLUSIONS: : The development of a method to visually demonstrate a rash on a mannequin facilitated some of the educational goals of the course: the importance of skin changes in aiding the diagnosis of acute anaphylaxis and the importance of exposure of the skin of the patient. PMID- 21646987 TI - Right ventricular infarction. PMID- 21646988 TI - Big society: real opportunity for integrated patient care. PMID- 21646989 TI - Lubrication and female catheterization. PMID- 21646990 TI - Fred's furious: social care must not be disregarded. PMID- 21646991 TI - Prisoner health: HIV infection and other blood-borne viral infections. AB - All prisoners should be entitled to the same standards of health care as the general population. This is the principle of equivalence. Inmates who are physically or mentally ill, addicted to substances, or disabled should receive equivalent care to what is expected of the NHS. There are a number of variations and inequalities in the health services that prisoners receive across the UK. A number of prisons have no policies in place for HIV, hepatitis C or sexual health. The prevalence of HIV is significantly higher in the prison population than it is in the wider community. This article considers and outlines the important principle of equivalence and the needs of the prison population in relation to HIV infection and other blood-borne virus infections. Methods used to minimize risk are described and the importance of development, implementation and evaluation of harm minimization strategies are discussed. The term 'prison' in this article is used to identify all places of detention in England and the term 'prisoner' is used to describe all adults who are held in such places. Prison services and healthcare provision in other UK countries are also outlined. PMID- 21646992 TI - Prisoner health: assessing a nurse-led hepatitis C testing clinic. AB - Prisons provide an ideal setting in which to carry out blood-borne virus testing among a high-risk population. This paper describes a nurse-led testing programme offered to all prisoners within the substance misuse unit at a local prison in South Wales over a 4-year period. Challenges to organizing such a clinic within the prison setting are discussed. Between April 2006 and April 2010, approximately 4500 prisoners were admitted through the substance misuse unit. Blood-borne virus testing took place among 176 prisoners, of which 24 were hepatitis C seropositive, 19 were confirmed as virus-positive, and eight were referred to specialist teams to consider treatment. Among prison populations blood-borne virus testing needs to be encouraged and improved. Offering testing provides opportunities for harm minimization, education and referrals for treatment. Such provisions are needed to benefit the health of individuals, their families and the wider public health. PMID- 21646993 TI - Knowledge, attitudes and practices in the provision of nutritional care. AB - The nutritional care of patients is one of the primary responsibilities of all registered nurses (Persenius et al, 2008). A poor nutritional status can lead to malnutrition, which can have serious consequences for an individual's quality of life (Field and Smith, 2008). This paper commences with an introduction to the concept of nutrition, provides an overview of nutritional guidelines and nutritional screening tools which identify those at risk of malnutrition. It reviews the literature on nurses' knowledge, attitudes and practices in the provision of nutritional care and debates challenges and opportunities encountered to help nurses ensure adequate patient nutrition. PMID- 21646994 TI - Reducing haemodialysis access infection rates. AB - Infections are the second most common cause of vascular access loss in the long term haemodialysis patient, and recent years have seen an increase in healthcare associated infections (HCAIs) associated with vascular access (Suhail, 2009). There have been a number of drivers including publication guidelines (Department of Health, 2006; 2007) and local protocols providing evidence-based recommendations that, when implemented, can reduce the risk of these infections. In England, the selection of bloodstream infections caused by methicillin resistant staphylococcus aureus (MRSA) as a significant clinical outcome has led to a vast amount of work in this area. Root cause analysis of individual infections (by the clinical teams when these occur) in many specialities identified areas where practice could be improved, including practice relating to vascular access within the renal setting. Manufacturers have also supported this work by focusing on developing products that are designed to reduce the likelihood of infections occurring. One product identified and used within the NHS is Chloraprep. PMID- 21646995 TI - Simulated practice--a panacea for health education? AB - A variety of approaches to teaching and learning are required to encompass the complexity of nursing. Simulated practice involves computer-based simulated environments, human patient simulation and role play, enabling multiple learning objectives to be taught in a realistic clinical environment without harming patients. The Nursing and Midwifery Council recognizes the use of simulated practice and has enabled higher education pre-registration nurse programme providers to incorporate a maximum of 300 hours of simulated practice as part of the 2300-hour practice hours component. However, not all students respond positively to simulated activity, which should be used to augment clinical practice and not replace it. It is important to ensure the quality of the simulation enables the educational experience to be positive for students. PMID- 21646996 TI - Reforming health research regulation: an incomplete solution. AB - Following the 2011 Budget, the UK Government has announced that as part of its reform of the NHS it will be establishing a new special health authority which will have the task of regulating research. This article considers these proposals and suggests that while a strong health regulator may be in itself a praiseworthy initiative, the current proposals simply do not go far enough and there is a need for radical reconceptualization of research regulation as a whole. PMID- 21646997 TI - COPD: criteria to assist in the identification of the palliative phase. AB - For people with chronic obstructive pulmonary disease (COPD), hospital admission can be associated with a poor prognosis. Consequently, the end stage of the illness needs to be recognized for timely palliative care to be initiated. Tools to enhance the palliative phase such as the Gold Standards Framework and the Liverpool Care Pathway rely on the recognition of the final phase of a person's life. The illness trajectory of cancer makes this recognition far easier than for COPD, as a result many patients and their families manage at home with limited support. Lincolnshire Respiratory Network has developed criteria to help recognize the end-stage COPD, which correlate well with recent recommendations from the Consultation on a Strategy for Services for COPD in England (Department of Health, 2010). However, there needs to be appropriate training to assist practitioners in their confidence to refer patients with end-stage COPD to palliative care providers. PMID- 21646998 TI - Nurses and midwives at the frontline of care. PMID- 21646999 TI - The global problem of adverse patient safety incidents in health care. AB - John Tingle discusses work carried out by the World Health Organization in patient safety and a report from the European Commission. PMID- 21647000 TI - Supporting self and others: from staff nurse to nurse consultant. Part 2: learning from experience. AB - This series of articles explores various ways of supporting staff who work in the fast-moving and ever-changing health service. In the first article, John Fowler, an experienced nursing lecturer, author and consultant examined the importance of developing a supportive working culture. Here in the second part of the series, he looks at the supportive effects of learning from experience. PMID- 21647001 TI - Nursing history: ships and sweets. AB - The following is a personal account of a nurse who escorted a patient with malaria home to England. It is an interesting story recounting the problems of visiting certain countries 100 years ago and the dangers it held at the time. PMID- 21647002 TI - The impact of the Bible on nursing. PMID- 21647003 TI - Cancer care: tackling the side effects. PMID- 21647004 TI - Chemotherapy-induced alopecia: advice and support for hair loss. AB - This article provides insight into the growth cycle of a hair follicle and the potential impact chemotherapy agents can have on this process, which often results in hair loss (alopecia). It explores the psychological consequences of chemotherapy-induced alopecia for an individual as a result of the perceptions of others as well as an individual's perception of his or her self-image. Despite the development of various forms of scalp cooling, chemotherapy-induced alopecia remains a major side effect for patients receiving chemotherapy; however, there have been improvements in wig provision and changing public opinion relating to baldness. Although chemotherapy-induced alopecia affects both males and females and all age groups, this article focuses on the potential impact for patients receiving chemotherapy as a form of treatment for breast cancer. As professionals we need to understand the social significance of hair in relation to a person's outward presentation and social interactions, along with the possible psychological implications of a person losing his or her bodily hair, and not just the head hair. We must aim to minimize the distress alopecia can cause by: ensuring we provide patients with up-to-date verbal and written information to enable them to prepare for losing their hair; helping them to preserve their self image and minimize the psychological consequences of hair loss while receiving chemotherapy; and preparing them for their hair re-growth following completion of chemotherapy. PMID- 21647005 TI - Managing renal cell carcinoma: the nurse's role. AB - Diagnosing renal cell carcinoma (RCC) can be difficult. Symptoms include a palpable mass in the abdomen, haematuria and hydronephrosis, but diagnosis can be delayed because of symptom similarities with other conditions, and often occurs incidentally during radiographic imaging for other complaints. This aside, more than 6600 people are diagnosed with kidney cancer annually in the UK (Patient UK, 2011). Although numerous improvements and greater use of screening measures have occurred, screening for this disease is limited to those patients identified as having a known genetic lineage linked to RCC (Pirass and Dahlstrom, 2010). PMID- 21647006 TI - Assessing the impact of breakthrough cancer pain. AB - Breakthrough pain is a transient exacerbation of pain that occurs either spontaneously or in relation to a specific predictable or unpredictable trigger despite relative stable and adequately controlled background pain. Breakthrough pain is a common and distinct component of cancer pain and is typically of rapid onset, severe in intensity, and generally self-limiting with an average duration of 30-60minutes. Despite the self-limiting nature of breakthrough pain, it can place significant physical, psychological, and economic burdens on both patients and their carers. Patients with breakthrough pain are often less satisfied with their analgesic therapy, they have decreased functioning because of their pain, and may also experience social and psychosocial consequences, such as increased levels of anxiety and depression. Successful management of breakthrough pain is best achieved by a thorough assessment which includes determining the severity, pathophysiology, and aetiology of the pain and takes into account both background and breakthrough pains while considering whether the underlying disease, co morbidities or precipitating events are amenable to interventions. The features of breakthrough pain and the challenges it presents to patients, their carers, and health professionals are illustrated with a case study. PMID- 21647007 TI - Should we recycle more than our plastic? PMID- 21647008 TI - Chemotherapy induced nausea and vomiting: a European perspective. AB - International guidelines have been available for the successful management of chemotherapy-induced nausea and vomiting (CINV) for several years, yet the experience of patients receiving chemotherapy continues to reflect variable symptom control. This article explores the clinical awareness and application of the guidelines across several European countries from the perspective of the nurses working in oncology units and centres who met at an expert European forum in London in June 2010. It will also identify the existing barriers to the use of best practice guidelines and some of the reasons why clinicians may not be aware of this guidance. There will be some discussion around the use of individualized risk assessments and the prescribing of the most appropriate antiemetic to meet the needs of the individual patient. It will also consider the hospital-produced algorithms and local drug protocols and the limitations on being able to deliver optimal symptom control from day one of cycle one. The role of the nurse in providing holistic care is key to making the experience for chemotherapy patients one to remember, not one they will never forget. PMID- 21647009 TI - CINV in the UK: a prospective audit of patient experience. AB - Chemotherapy induces nausea and vomiting in a significant number of cancer patients across the world each day. Unfortunately patients tend to be at home and away from the specialist oncology centres when the symptoms are at their worst. This leads to a disconnect between what health professionals predict will happen to patients and what patients actually experience, especially in the delayed phase of chemotherapy-induced nausea and vomiting (CINV) (Grunberg, 2004). Following feedback from a European forum (see preceding article, p. 22-8) that CINV is still poorly managed, the UK Oncology Nursing Society (UKONS) is proposing a nation-wide, multi-site audit of the patient-reported CINV experience by reviewing specially-designed CINV diaries. Each cycle of treatment will be recorded and side effects closely monitored for several days after treatment. Furthermore, the audit will assess the clinical prescribing habits, use of international CINV guidelines and variations in local protocols for these patients. An online survey will be undertaken by the clinical teams on what they predict a group of chemotherapy patients will experience by way of CINV. This audit will then be compared to the reported side effects experienced by the patients. It is proposed that this audit will provide evidence of how NHS and private oncology teams manage CINV within the UK. PMID- 21647010 TI - NHS reforms: cutting where it counts. PMID- 21647011 TI - Stoma appliances: it's all about the bag! PMID- 21647012 TI - Renal cell carcinoma: treatment options. PMID- 21647013 TI - Isolation prioritization tool: revision, adaptation and application. AB - The provision of single rooms for the care of patients who require isolation may not match the number required. Placing patients in isolation facilities may have an effect on their psychological wellbeing and the quality of care delivered. To ensure a rational and consistent approach to the prioritization of single room usage, an existing prioritization system was revised. This was validated by a group of experts and tested in an acute hospital. A simple short cut guide was developed and used. PMID- 21647014 TI - An aging renal population-is dialysis always the answer? AB - As people age and live for longer they are more likely to develop comorbid conditions including chronic kidney disease (CKD). This paper discusses the treatment options for stage 5 CKD including haemodialysis, peritoneal dialysis, transplantation or conservative management, also known as supportive care, for those who decide not to undertake dialysis. It also highlights the complexity of offering a treatment such as dialysis, viewed as a requirement to prolong life, without which people will die, which is unable to restore the kidneys to normal function, only substitute for. Dialysis is also an arduous therapy known to shorten life. In the past refusal of dialysis was viewed as akin to suicide and it is not until more recently that the needs and experiences of those who decide not to embark on dialysis have started to be recognized. Clearly dialysis is not suitable for all, particularly those who are frail with multiple comorbidities and so supportive and palliative care may be a more suitable option for some. PMID- 21647015 TI - Parkinson's: symptoms, treatments and research. AB - Parkinson's is a incurable progressive neurological condition caused by a degeneration of dopamine-producing cells. The most common symptoms of the condition include slowness of movement, tremor and muscle stiffness; other symptoms include sleep difficulties, depression and anxiety. Parkinson's is usually controlled by medication but other treatments and therapies, such as occupational therapy, physiotherapy and speech and language therapy, can also be effective in controlling and managing symptoms. The timing of this medication is key to the successful management of Parkinson's. Delays to initiating medication or missed doses can have serious consequences, and it can take some time for symptoms to stabilize. Research into treatments for Parkinson's and, ultimately, finding a cure for the condition, is ongoing. Parkinson's UK researchers are working on projects to identify the causes of cell death and tests to detect the condition at its earliest possible stage. Current treatments can help to ease the symptoms but none can repair the damage in the brain or slow the progress of the condition; now, Parkinson's UK researchers are working to develop new treatments that can. PMID- 21647016 TI - Childhood sexual abuse: help make it 'Safe to Say'. AB - Childhood sexual abuse remains a taboo subject and requires sensitive handling by committed and caring services (Scottish Executive, 2004). By introducing a training programme, Safe to Say, the Scottish Association of Mental Health aims to improve the confidence and competence of healthcare workers when faced with the sensitive disclosure of childhood abuse. This article aims to explore the effects of childhood sexual abuse and the attitudes of health professionals when faced with adult survivors disclosing such abuse. Despite Government strategies, there still appears to be a lack of suitable services for adult survivors of childhood sexual abuse, even though health professionals have an essential role in supporting disclosure and promoting recovery (Read et al, 2005). PMID- 21647017 TI - Effects of patient death on nursing staff: a literature review. AB - There were 509090 deaths recorded in England and Wales for 2008 (Office for National Statistics, 2010); of these, over 56% (260000) occurred in NHS hospitals. The death of a patient is an event that most, if not all, nursing staff will encounter during their work. This experience can elicit physical, cognitive, behavioural, spiritual and emotional responses (Parkes, 1998). AIM: The aim of this literature review is to explore how the death of patients in a hospital setting impact on nursing staff. METHOD: A review of the literature was undertaken using the online databases CINAHL, Medline and PsychInfo. The search was limited to articles in the English language and those from peer-reviewed journals. RESULTS: Themes arising from the literature review included: the theoretical context; the emotional impact; the culture of the healthcare setting; staff's previous life experiences; and support available for healthcare staff. CONCLUSIONS: The death of patients does have an impact on nurses. This can affect them both in their work environment and outside of work. Education around grief theory and support from others are helpful for staff in developing strategies for coping with patient deaths. PMID- 21647018 TI - Indwelling urinary catheters: accurate monitoring of urine output. AB - Caring for patients who have an indwelling urinary catheter is a common feature of clinical nursing practice in the acute care environment, which for some patients is essential to their medical management. However, evidence suggests that in many cases catheterization may be unjustified, exposing the patient to a significant risk of acquiring a urinary tract infection, which extends length of stay and imposes an extra burden of care and cost. Improving the care for these patients depends on good education, implementation of best practice, adequate resources, and effective devices and treatment. This article focuses on a clinical, unbiased review of a new product for the accurate monitoring of urine output. PMID- 21647019 TI - Middle-aged women and everyday life: implications for health. AB - The aim of this study was to describe middle-aged women's perceptions of lifestyle and how it was experienced in their daily lives. Interviews with 15 women selected using network sampling were analysed using a phenomenographic approach. Lifestyle was perceived as either 'handle life', describing a framework of how everyday life should manifest itself in different areas of life, or 'relate to life' implying accepting life as it is and making the best of it based on previous experiences. In terms of providing preventive and health-promoting guiding, it is essential that nurses have an insight into people's reasoning to support them to improve health or deal with illness. PMID- 21647020 TI - What do NHS staff think about working in the NHS? AB - John Tingle discusses some issues highlighted by the 2010 NHS Staff Survey in England. PMID- 21647021 TI - Supporting self and others: from staff nurse to nurse consultant. Part 1: chocolate and Marmite! AB - This series of articles explores various ways of supporting staff who work in the fast-moving and ever-changing health service. In this first article, John Fowler, an experienced nursing lecturer, author and consultant, examines the importance of developing an informal working culture of support from both the individual's perspective and from the perspective of the health service management. PMID- 21647022 TI - Improving the NHS in a climate of austerity. PMID- 21647023 TI - Why are nurse mentors failing to fail student nurses who do not meet clinical performance standards? AB - This article reports findings from a qualitative study exploring mentors' reluctance to refer students who did not perform adequately in clinical settings. A focus group and individual semi-structured interviews were undertaken with experienced stage two mentors in an NHS trust. Mentors spoke of having to balance objectivity and intuition when dealing with upset or angry students, those who were popular or those who tried hard but did not perform adequately. They describe a number of factors that led them to pass a student who was less than satisfactory. In particular the findings highlight difficulties with measuring and recording attitudes. The data demonstrate the willingness and motivation of mentors to keep up to date and learn from their students. However, findings also suggest the need for mentors to have greater confidence in their own clinical abilities. Recommendations are made for training strategies to address these barriers in order to ensure that only competent, caring nurses are allowed to register. PMID- 21647024 TI - The keys to success as a nurse. PMID- 21647025 TI - The relevance of Nightingale today. PMID- 21647032 TI - How to write a surgical operation note. PMID- 21647033 TI - Imaging the liver: use of different modalities. PMID- 21647034 TI - Management of acute epiglottitis in adults. PMID- 21647035 TI - Abstracts from the UK Medical Students' Association conference. PMID- 21647036 TI - Importance of the hospital autopsy in clinical governance, teaching and research. PMID- 21647037 TI - Supporting tomorrow's doctors. PMID- 21647039 TI - Acute aortic syndrome: surgical, endovascular or medical treatment. PMID- 21647040 TI - Do not attempt resuscitation orders, ethics and the Mental Capacity Act. PMID- 21647041 TI - Non-traumatic neurological emergencies presenting to the intensive care unit. PMID- 21647042 TI - Teleradiology: 21st century communication in surgery. PMID- 21647043 TI - The athlete's heart. PMID- 21647044 TI - Medical leadership: from policy to practice. PMID- 21647045 TI - August Bier: father of spinal and regional anaesthetic blocks. PMID- 21647046 TI - Endovascular treatment of a secondary aortoenteric fistula: a temporary solution? PMID- 21647047 TI - Caecal perforation following a caesarean section. PMID- 21647048 TI - Intrahepatic abscess secondary to acute calculous cholecystitis. PMID- 21647049 TI - Amnesia as a sex-related adverse event. PMID- 21647050 TI - Role of computed tomography in upper gastrointestinal bleeding. PMID- 21647051 TI - Entitlement to hospital treatment in the UK. PMID- 21647054 TI - Role of prone ventilation in the management of patients with acute respiratory distress syndrome. PMID- 21647064 TI - Petition for change. PMID- 21647065 TI - An exploration of nursing documentation of pressure ulcer care in an acute setting in Ireland. AB - OBJECTIVE: To explore the nature and quality of documented care planning for pressure ulcers in a large teaching hospital in the Republic of Ireland. METHOD: A mixed method design was used; this encompassed a descriptive survey that retrospectively evaluated nursing records (n=85) in two wards (orthopaedic and care of the older adult) and a focus group (n=13) that explored nurses' perspectives of the factors influencing concordance and the quality of nursing documentation. Only records of at-risk patients (Waterlow score of >10) were included. RESULTS: It was identified that 47% (n=40) were assessed as at high or very high risk of developing a pressure ulcer. Fifty-two patients (61%) had a weekly risk assessment, but 25% (n=21) had only one follow-up assessment. Only 45% (n=38) of charts had some evidence of documented care planning, and of those 53% (n=20) had no evidence of implementation of the care plan and 66% (n=25) had no evidence of outcome evaluation. Only 48% (n=41) of this at-risk population was nutritionally assessed. Of patients admitted with and without a pressure ulcer, there was no record of regular positioning in 70% (n=59) and 60% (n=51) respectively. CONCLUSION: Documentation on pressure ulcer care is not standardised and requires development. CONFLICT OF INTEREST: None. PMID- 21647066 TI - Use of granulated sugar therapy in the management of sloughy or necrotic wounds: a pilot study. AB - OBJECTIVE: To determine the in vitro antimicrobial efficacy of three types of sugar and conduct a pilot clinical study with a view to developing a protocol for a randomised controlled trial (RCT). METHOD: In the in vitro studies three types of granulated sugar (Demerara, granulated beet sugar and granulated cane sugar) were tested to determine their minimum inhibitory concentrations (MICs) against 18 Gram-negative and Gram-positive bacteria in a micro-titre broth dilution assay; growth inhibition of Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa in different concentrations of sugar (0.38-25%) was also tested over 12-hours in an agar diffusion assay. The pilot clinical study selected patients from a vascular surgical ward and a vascular outpatient department. All had acute or chronic exuding wounds, some of which were infected. White granulated sugar was applied to the wounds. The following parameters were assessed: surface area; wound characteristics including pain, malodour, appearance (slough/granulation); exudate level; pain level and bacterial load. Patients with diabetes had their blood sugar levels checked daily. All patients completed a short health questionnaire at the start and end of the study. Staff completed a satisfaction questionnaire at the end of the study. The study period was 21 days. RESULTS: In vitro tests demonstrated that sugar inhibits bacterial growth. All three types of sugars had MICs ranging from 6-25% in the bacterial strains tested. The diffusion tests showed that strains were able to grow well in low concentrations of sugar but were completely inhibited in higher concentrations. The two granulated sugars were found to be slightly more effective than Demerara sugar, so the latter was excluded from the clinical pilot study. Twenty-two patients (20 inpatients and two outpatients) with sloughy or necrotic wounds were recruited into the clinical study. Two patients had MRSA and two had Staphylococcus colonisation at baseline. Blood sugar levels remained stable in the seven patients with insulin-dependent diabetes mellitus. All wounds were clean/debrided in a mean of 11.13 days. Pain and malodour reduced markedly. Patient and staff surveys revealed overwhelming support for the sugar therapy. CONCLUSION: The pilot study achieved its aim of developing a protocol for a RCT. Preliminary data suggest that sugar is an effective wound cleansing and is safe to use in patients with insulin-dependent diabetes. In vitro studies demonstrate that sugar inhibits bacterial growth. CONFLICT OF INTEREST: None. PMID- 21647067 TI - Biofilms and their management: from concept to clinical reality. AB - How can a better understanding of wound biofilms change your clinical practice? And what is the likely impact on clinical outcomes? At this year's JWC lecture, sponsored by sorbion, two of the world's leading microbiologists, Steve Percival and Randy Wolcott, answered these questions. Steve Percival explained what a wound biofilm is, and what diagnostic and management techniques can be used to overcome it. Randy Wolcott described the astonishing success this can bring, with healing rates of up 90% being recorded in patients with recalcitrant chronic wounds. PMID- 21647068 TI - Molecular diagnostics and personalised medicine in wound care: assessment of outcomes. AB - OBJECTIVE: This large, level A, retrospective cohort study set out to compare healing outcomes in three large cohorts of wound patients managed universally for bioburden: standard of care group, who were prescribed systemic antibiotics on the basis of empiric and traditional culture-based methodologies; treatment group 1, who were prescribed an improved selection of systemic antibiotics based on the results of molecular diagnostics; treatment group 2 who received personalised topical therapeutics (including antibiotics) based on the results of molecular diagnostics. METHOD: Apart from the differences in diagnostic methods and antibiotic treatments described above, all three cohorts were subjected to the same biofilm-based wound care protocol, which included evaluation of the host and bioburden, frequent sharp debridement, use of wound dressings and comprehensive standard care (reperfusion therapy, nutritional support, offloading, compression and management of comorbidities). RESULTS: In all, 1378 patients were recruited into the study. In the standard of care group 48.5% of patients (244/503) healed completely during the 7-month study period. This increased to 62.4% (298/479) in treatment group 1 and 90.4% (358/396) in treatment group 2. Cox proportional hazards analysis revealed the time to complete closure decreased by 26% in treatment group 1 (p<0.001) and 45.9% in treatment group 2 (p<0.001) compared with the standard of care group. Patients in treatment group 2 had >200% better odds of healing at any given time point compared with the other cohorts. CONCLUSION: Implementation of personalised topical therapeutics guided by molecular diagnosis resulted in statistically and clinically significant improvements in outcome. The integration of molecular diagnostics and personalised medicine provides a directed and targeted approach to wound care. CONFLICT OF INTEREST: SED and RDW are owners of PathoGenius Laboratories, a clinical diagnostic laboratory. SED and RDW are owners of Research and Testing Laboratory, which develops molecular diagnostics. CJ and JK are clinical advisors for PathoGenius. CJ and JK are owners of Southeastern Medical Compounding, Savannah, GA and Southeastern Medical Technologies, Savannah, GA. PMID- 21647069 TI - Clinical efficacy of a new monofilament fibre-containing wound debridement product. AB - OBJECTIVE: To evaluate the wound debridement efficacy (that is, achievement of 100% granulation tissue on the wound bed) of a new monofilament fibre product (Debrisoft). METHOD: This multicentre, prospective, observational evaluation assessed the debridement efficacy, safety, patient comfort and user satisfaction of this new product. Time taken to perform the debridement procedure was also recorded. The new product was wetted with either saline or polihexanide and applied for 2-4 minutes, following which the usual dressing regimen was applied. Clinical outcome was scored by a trained clinician. Additionally, before and after photographs were assessed by one and the same clinician, who was blinded to the treatment given. The debridement outcomes achieved with the test product were compared with results obtained using other methods of debridement, both non surgical and surgical, taken from an electronic database but using the same scoring systems as here. RESULTS: Sixty patients with chronic wounds requiring debridement were recruited, of whom 57 were included in the analysis. Debridement was effective in 93.4% (142/152) of the sessions, and the product remained intact in 95.4% (145/152). The average time for each debridement session was 2.51 minutes, markedly less than for the current debridement methods at the evaluation centres. Visible debris and slough were successfully removed with the test product. Patients reported no pain during the procedure in 45% of cases and slight discomfort for a short duration (2.0 minutes on average) in 55% of cases. CONCLUSION: The results indicate the potential for this monofilament fibre product to replace several modes of debridement, based on its efficacy, short procedure, ease of use and patient comfort. CONFLICT OF INTEREST: The evaluation protocol was proposed and supported by Lohmann & Rauscher GmbH, who provided the evaluation products. MS and MA are employees of Lohmann & Rauscher. The other authors declare to have no relevant financial interest in the evaluation. Apart from input to the protocol, the sponsors had no role in the conduct of the study, such as data collection, analysis, or preparation, review, or approval of the manuscript. PMID- 21647070 TI - Defining a holistic pain-relieving approach to wound care via a drug free polymeric membrane dressing. AB - Wound care practice continuously demonstrates that healing cannot be adequately controlled if a patient's experience of pain is not managed effectively. Current pain management guidelines do not account for the holistic treatment of pain emanating from a wound-an environment of uncontrolled or rogue inflammation, neuropathy and neuroischaemia. This article investigates how polymeric membrane dressings can interact with the pathology of wounds to correct abnormalities in pain pathways of the nervous system and dampen problematic ongoing pain to enhance the clinical picture of wound healing. PMID- 21647073 TI - Bringing death further into the open. AB - This month, BBC1 screened a television programme, Inside the Human Body: First to Last, in which a man's death was shown. This event was apportioned only a fraction of the total running time, but it garnered a lot of attention in the media. What did the programme, and the reactions of the national press and general public, tell us about current attitudes towards death and dying? PMID- 21647074 TI - The pursuit of cultural competence: service accessibility and acceptability. AB - Inequalities in end-of-life care provision are well documented and are a priority for organizations both nationally and internationally. It is well recognized that end-of-life care should be provided not just for patients with cancer, and that access to services should be based on need rather than on disease. Of even greater importance, particularly in multicultural societies, is the realization that such services should also be accessible to all, regardless of gender, culture, religion, ethnicity, etc. To achieve this, a degree of cultural sensitivity across care provision sectors is a prerequisite. Services must be acceptable as well as accessible to all, regardless of ethnic or cultural background. PMID- 21647075 TI - Principles of nutrition in the palliation of long-term conditions. AB - Palliative care, directed toward alleviating symptoms and improving quality of life, should be available to all those with chronic or incurable illness. Good nutrition is central to such care meeting patients' physical requirements and psychosocial needs, thus helping them to maintain a sense of autonomy and normality while also promoting psychological comfort, communication, and social interaction. Many patients receiving palliative care experience difficulties in maintaining an appropriate food intake, which presents many challenges for practitioners and requires an understanding of both nutrition and patient needs. This article describes the types of nutritional support that are available in the palliative care setting, as well as important ethical factors relevant to considerations of starting and withdrawing artificial nutrition. PMID- 21647076 TI - Palliative care clinical trials: how nurses are contributing to integrated, evidence-based care. AB - The aim of this paper is to describe the emerging role of the palliative care clinical trials nurse in an era of evidence-based practice and increasing clinical trial activity in palliative care settings across Australia. An overview of the current clinical trials work is provided, with a focus on three aspects of clinical trials nursing practice that have significant implications for patients: managing the consent process, integrating clinical trials into multidisciplinary care, and establishing and building the evidence base to inform practice in palliative care settings. Clinical trials roles provide palliative care nurses with an opportunity to contribute to clinical research, help expand palliative care's evidence base, and develop their own research capabilities. PMID- 21647077 TI - Collaborative lymphoedema management: developing a clinical protocol. AB - Lymphoedema is a very distressing chronic condition prevalent in some metastatic cancers. Conservative treatment of lymphoedema in palliative care involves complete/complex decongestive therapy (CDT) using manual lymphatic drainage (MLD), compression therapy (bandaging and/or garments), skincare, and remedial exercises, adapted to the needs of the patient. The aim of this service development project was to identify current practice in a hospice palliative care service, develop new assessment tools, and implement a collaborative clinical protocol to improve access to lymphoedema management for patients in the hospice. Two audits provided new evidence about patient profiles, patient assessment, and treatment outcomes for cancer- and non-cancer-related lymphoedema. This project had a quality-improvement effect on service delivery and developed an effective partnership approach to lymphoedema management between local district nursing services and the specialist lymphoedema physiotherapist. PMID- 21647078 TI - Nurses' views on using the Liverpool care pathway in an acute hospital setting. AB - The Liverpool Care Pathway (LCP) is one of three major recent UK initiatives to improve the care of people dying with malignant and non-malignant disease. This focus group research aimed to explore nurses' perceptions of using the LCP in an acute hospital setting, their perceptions of its impact on patient care, and their confidence in care at the end of life, as well as to generate new ideas among peers. Six themes were identified: use of the LCP as a care tool; improved confidence, training, and support; the nursing-medical relationship; dying in hospital; and end-of-life issues. The nurses reported that the LCP has increased their confidence and resulted in improved patient care. Exploring the effect of the LCP on patient care may provide valuable data that may inform how the LCP is integrated into practice in the future. PMID- 21647079 TI - The constraints of the ordinary: 'being with' in the context of end-of-life nursing care. AB - The concept of 'being with' was an essential part of the hospice philosophy developed by Cicely Saunders, and one that she believed was a key element in the care of the dying. In Saunders' philosophy, 'being with' is a specific moment of engagement between a health professional and a patient whereby the patient has the opportunity to examine and express their thoughts and feelings relating to their imminent death. This study examined how 'being with' was implemented as a part of ordinary day-to-day practice in a hospice in Scotland. The study was based on the principles of ethnomethodology, which aims to reveal embedded and taken-for-granted rules and behaviours that determine practice. The data collection and analysis revealed that the nursing practice was constructed in such a way that 'being with', as Saunders understood it, could not be initiated either by a member of the nursing staff or by the patients. By adopting the philosophical framework of Heidegger, the study offers an understanding of how the nursing staff had developed 'being with' as provision of comfort in order to protect both themselves and the patients from engaging with painful emotions associated with facing death. PMID- 21647080 TI - Preferences for end of life: views of hospice patients, family carers, and community nurse specialists. AB - An exploratory qualitative study was undertaken with the aim of identifying issues around discussing and recording preferences on place of death from the perspective of hospice patients, carers, and hospice community nurses. A purposive sample was selected from three hospices in the south east of England of six community nurse specialists (CNSs), five patients, five carers, and five bereaved carers. The patients and carers said it was important to them to discuss their preferences, but they acknowledged that discussing dying was difficult. The CNSs stated that there were several barriers that inhibited end-of-life discussions and that discussing preferences was not always a primary goal of care planning. It was felt among the service users, particularly the bereaved carers, that having more knowledge about what to expect of the dying process, knowing their relative's wishes, and understanding the role of hospice and palliative care could improve the experience of events leading up to death. PMID- 21647082 TI - Politics and palliative care: Ireland. AB - Eire, or in its English language form, Ireland, is a constitutional republic island state bordered by the Atlantic Ocean on its rugged north and rocky west coasts and the rough Irish Sea on the east. After a period of calm, financial promise, and prosperity, the so called Celtic tiger's roar is less pronounced at the moment. A recent change in the Irish government heralds a period of austerity after a decade or more of almost double-digit growth. PMID- 21647087 TI - Social inequalities in alcohol-related adult mortality by National Statistics Socio-economic Classification, England and Wales, 2001-03. AB - BACKGROUND: This article is the first analysis of the social inequalities in adult alcohol-related mortality in England and Wales at the start of the 21st century, using the National Statistics Socio-economic Classification (NS-SEC). It presents the socio-economic patterns of alcohol-related mortality by gender, age and region, for England and Wales as a whole, Wales and the regions of England. METHODS: Death registrations provided the number of deaths for working age adults, using the National Statistics definition of alcohol-related mortality. Population estimates for England and Wales in 2001-03 were used to estimate alcohol-related mortality rates by sex, five-year age group, NS-SEC and region. Inequalities were measured using ratios of alcohol-related mortality rates between the least and most advantaged classes. RESULTS: There were substantial socio-economic variations in adult alcohol-related mortality, with the inequalities being greater for women than for men. The mortality rate of men in the Routine class was 3.5 times those of men in Higher and Managerial occupations, while for women the corresponding figure was 5.7 times. Greater socio-economic inequalities in mortality were observed for men aged 25-49 than for men aged 50-64; however the highest mortality rate of men occurred for Routine workers aged 50-54. Women in the Routine class experienced mortality rates markedly higher than other classes. The highest mortality rate of women also occurred for Routine workers, but at a younger age than for men (45-49). Within England, the North-West showed the largest inequalities, with particularly high rates in the Routine class for both sexes. In general, there was no association between levels of mortality and socio-economic gradients in mortality across the English regions and Wales. CONCLUSIONS: Rates of alcohol-related mortality in England and Wales increased significantly for people between the early 1990s and early 21st century, and were substantially greater for those in more disadvantaged socio-economic classes. There is also evidence that these socio-economic differences were greater at younger ages, especially for men at ages 25-49. PMID- 21647088 TI - Disability-free life expectancy: comparison of sources and small area estimates in England, 2006-08. AB - BACKGROUND: Health expectancies (HEs) at sub-national geographies or by clusters of areas defined by relative deprivation are important tools to monitor inequalities in health. Previously, analyses have had limited usefulness due to a lack of timeliness or local relevance caused by limitations on the frequency or coverage of survey data. Here we explore the potential of the Annual Population Survey (APS) to provide robust estimates of disability-free life expectancy (DFLE) for men and women by clusters of area deprivation, English regions and local authority districts (LAs) in the period 2006-08. METHODS: DFLE estimates for the UK were compared using the prevalence of limiting long-standing illness (LLSI) calculated using data from the APS and from the General Lifestyle Survey (GLF) covering Great Britain and equivalent data from the Continuous Household Survey (CHS) covering Northern Ireland, aggregated over the period 2006-08. The further use of APS data for England enabled the calculation of estimates of DFLE at age 16 and at age 65 for men and women by area deprivation quintiles (each quintile comprises a fifth of areas ranked according to their relative deprivation), English regions and LAs in order to measure inequality in DFLE between these population groupings. RESULTS: The prevalence of LLSI and estimates of DFLE at national level were broadly comparable using APS and GLF/CHS data. Substantial inequality in DFLE was present between clusters of areas defined by relative deprivation and between English regions and LAs. The scale of inequality increased markedly with each finer geographical scale analysed. CONCLUSION: The APS is a viable data source to provide LLSI data for use in DFLE estimation across a range of areas and clusters of area deprivation. While increasingly fine grained analysis decreases statistical precision, it is possible to detect clear differences between areas within regions and in making comparisons with the England average. The estimates presented here provide scope to set benchmarks for assessing the impact of interventions designed to reduce inequality in DFLE beyond the period 2006-08. PMID- 21647089 TI - An analysis of the socio-demographic characteristics of sole registered births and infant deaths. AB - BACKGROUND: Differences in birth characteristics and infant mortality rates by marital status and birth registration type reflect complex underlying factors. In particular, births registered solely by the mother are seen as a disadvantaged group. This article analyses the socio-demographic characteristics of births by registration type and parents' marital status and explores these differences for health outcomes. METHODS: Birth notifications data from the NHS Numbers for Babies system was linked to birth registration data held by the Office for National Statistics (ONS) for births occurring in 2007 and 2008 in England and Wales. This dataset was further linked to death registration data to identify infants who died before their first birthday.Regression analysis was used to compare factors and health outcomes across marital and registration status groups. Regression models were calculated to determine the main risk factors for poor outcomes. RESULTS: The registration groups differed in the age of the mother at birth, the proportion of young mothers, ethnic group distribution and measures of deprived circumstances. The joint registered-different address and sole registered groups were similar in the proportion of young mothers and the deprivation indices. The groups also differed in the proportion of low birthweight and premature babies. The joint registered-different address and sole registered groups both had higher percentages of 'small for gestational age' babies compared with the within-marriage and joint registered-same address groups. The stillbirth rate was highest in the sole registered group. Both the joint registered-different address and sole registered groups had higher infant mortality rates compared with the within-marriage and joint registered-same address groups. Multivariate analysis indicated that low birthweight was a key factor in infant mortality. CONCLUSIONS: Births registered solely by the mother were found to be a disadvantaged group but were also similar to the joint registered group living at different addresses in their main socio-demographic characteristics and health outcomes. The joint-registered group living at the same address was similar to the within-marriage group across the same measures. This argues that, in the 21st century, the distinction between infants with 'resident' and 'nonresident' fathers is more meaningful for health outcomes than that between births inside and outside marriage. PMID- 21647090 TI - Diabetes causes multiple genetic alterations and downregulates expression of DNA repair genes in the prostate. AB - The molecular impact of diabetes mellitus on prostate gland has not been elucidated. In this study, we performed a whole-genome cDNA microarray analysis using a streptozotocin-induced diabetic rat model to identify the effects of diabetes on the gene expression profiles in prostate. Our study shows that diabetes causes changes in the expression of multiple genes, particularly those related to cell proliferation and differentiation, oxidative stress, DNA damage repair, cell cycle checkpoints, angiogenesis and apoptosis. These findings were confirmed by real-time polymerase chain reaction and immunohistochemical staining using rat and human prostate tissue. We also used a cell culture model (human normal prostatic RWPE-1 cell line) to study the direct effect of high glucose. We found that high glucose caused increased intracellular oxidative stress and DNA damage, as well as downregulation of anti-oxidative enzymes and DNA damage repair genes MRE11 and XRCC3. Our findings provide important insights into understanding the pathogenesis of the diabetes-induced changes in prostate as well as identifying potential therapeutic targets for future studies. PMID- 21647092 TI - Inhibition of active autophagy induces apoptosis and increases chemosensitivity in cholangiocarcinoma. AB - Intrahepatic cholangiocellular carcinomas (ICCs) are usually fatal neoplasms originating from bile duct epithelia. However, many cholangiocarcinoma cells are shown to be resistant to chemotherapeutic drugs, which induce cell apoptosis. The role of autophagy and the therapeutic value of autophagy-associated genes are largely unknown in ICC. Here, we showed that autophagy was activated in nutrient starvation and xenograft cholangiocarcinoma cells. Furthermore, expression of autophagic genes and their autophagic activity were higher in clinical ICC specimens than that in normal cholangiocytes separated by laser capture microdissection. Inhibition of autophagy by autophagy inhibitors or siRNA, cholangiocarcinoma cells showed detention of proliferation and increase of apoptosis during nutrient starvation. In addition, autophagy inhibitor treatment or knockdown of beclin 1 suppressed tumor growth and sensitized ICC cells to chemotherapeutic agent-induced cell death. In conclusion, our data showed that autophagy is activated in ICC, and inactivation of autophagy may lead to cell apoptosis and enhance chemotherapy sensitivity. PMID- 21647121 TI - When off label is off target. PMID- 21647122 TI - Market overlap points to irresponsible use of tuberculosis drugs. PMID- 21647123 TI - Disputed EU herbal medicine rules take force. PMID- 21647091 TI - The mitochondrial paradigm for cardiovascular disease susceptibility and cellular function: a complementary concept to Mendelian genetics. AB - While there is general agreement that cardiovascular disease (CVD) development is influenced by a combination of genetic, environmental, and behavioral contributors, the actual mechanistic basis of how these factors initiate or promote CVD development in some individuals while others with identical risk profiles do not, is not clearly understood. This review considers the potential role for mitochondrial genetics and function in determining CVD susceptibility from the standpoint that the original features that molded cellular function were based upon mitochondrial-nuclear relationships established millions of years ago and were likely refined during prehistoric environmental selection events that today, are largely absent. Consequently, contemporary risk factors that influence our susceptibility to a variety of age-related diseases, including CVD were probably not part of the dynamics that defined the processes of mitochondrial nuclear interaction, and thus, cell function. In this regard, the selective conditions that contributed to cellular functionality and evolution should be given more consideration when interpreting and designing experimental data and strategies. Finally, future studies that probe beyond epidemiologic associations are required. These studies will serve as the initial steps for addressing the provocative concept that contemporary human disease susceptibility is the result of selection events for mitochondrial function that increased chances for prehistoric human survival and reproductive success. PMID- 21647124 TI - New diabetes drugs go beyond insulin to flush out excess sugar. PMID- 21647125 TI - NIH funding rates drop to record lows. PMID- 21647126 TI - HPV protection in older groups may be in the eye of the beholder. PMID- 21647127 TI - Gene tests for brain injury still far from the football field. PMID- 21647128 TI - Jewish genetic screening grows despite questions about breadth. PMID- 21647129 TI - Attending conferences... under the influence. PMID- 21647130 TI - Chile plans new agency to boost pharma oversight. PMID- 21647131 TI - Drug companies hope to breathe life into asthma pipeline. PMID- 21647132 TI - On thirtieth anniversary, calls for HIV cure research intensify. PMID- 21647134 TI - Heat shock and awe. PMID- 21647135 TI - Caught on film. PMID- 21647136 TI - Persistence may pay off for antibiotics innovators. PMID- 21647137 TI - Straight talk with...Joel Kupersmith. Interview by Elie Dolgin. PMID- 21647138 TI - China needs to boost funding for graduate students to stay competitive. PMID- 21647140 TI - The holy grail of high bone mass. PMID- 21647141 TI - A CUE hints at tumor resistance. PMID- 21647142 TI - ERasing breast cancer resistance through the kinome. PMID- 21647143 TI - Imaging transplant rejection: a new view. PMID- 21647144 TI - Wormless without wingless. PMID- 21647145 TI - PGRPs kill with an ancient weapon. PMID- 21647146 TI - Tracing the roots of squamous cell carcinomas. PMID- 21647148 TI - Risk variant for schizophrenia in the neurogranin gene impacts on hippocampus activation during contextual fear conditioning. PMID- 21647149 TI - A multi-tissue analysis identifies HLA complex group 9 gene methylation differences in bipolar disorder. AB - Epigenetic studies of DNA and histone modifications represent a new and important activity in molecular investigations of human disease. Our previous epigenome wide scan identified numerous DNA methylation differences in post-mortem brain samples from individuals affected with major psychosis. In this article, we present the results of fine mapping DNA methylation differences at the human leukocyte antigen (HLA) complex group 9 gene (HCG9) in bipolar disorder (BPD). Sodium bisulfite conversion coupled with pyrosequencing was used to interrogate 28 CpGs spanning ~700 bp region of HCG9 in 1402 DNA samples from post-mortem brains, peripheral blood cells and germline (sperm) of bipolar disease patients and controls. The analysis of nearly 40 000 CpGs revealed complex relationships between DNA methylation and age, medication as well as DNA sequence variation (rs1128306). Two brain tissue cohorts exhibited lower DNA methylation in bipolar disease patients compared with controls at an extended HCG9 region (P=0.026). Logistic regression modeling of BPD as a function of rs1128306 genotype, age and DNA methylation uncovered an independent effect of DNA methylation in white blood cells (odds ratio (OR)=1.08, P=0.0077) and the overall sample (OR=1.24, P=0.0011). Receiver operating characteristic curve A prime statistics estimated a 69-72% probability of correct BPD prediction from a case vs control pool. Finally, sperm DNA demonstrated a significant association (P=0.018) with BPD at one of the regions demonstrating epigenetic changes in the post-mortem brain and peripheral blood samples. The consistent multi-tissue epigenetic differences at HCG9 argue for a causal association with BPD. PMID- 21647150 TI - Modeling the functional genomics of autism using human neurons. AB - Human neural progenitors from a variety of sources present new opportunities to model aspects of human neuropsychiatric disease in vitro. Such in vitro models provide the advantages of a human genetic background combined with rapid and easy manipulation, making them highly useful adjuncts to animal models. Here, we examined whether a human neuronal culture system could be utilized to assess the transcriptional program involved in human neural differentiation and to model some of the molecular features of a neurodevelopmental disorder, such as autism. Primary normal human neuronal progenitors (NHNPs) were differentiated into a post mitotic neuronal state through addition of specific growth factors and whole genome gene expression was examined throughout a time course of neuronal differentiation. After 4 weeks of differentiation, a significant number of genes associated with autism spectrum disorders (ASDs) are either induced or repressed. This includes the ASD susceptibility gene neurexin 1, which showed a distinct pattern from neurexin 3 in vitro, and which we validated in vivo in fetal human brain. Using weighted gene co-expression network analysis, we visualized the network structure of transcriptional regulation, demonstrating via this unbiased analysis that a significant number of ASD candidate genes are coordinately regulated during the differentiation process. As NHNPs are genetically tractable and manipulable, they can be used to study both the effects of mutations in multiple ASD candidate genes on neuronal differentiation and gene expression in combination with the effects of potential therapeutic molecules. These data also provide a step towards better understanding of the signaling pathways disrupted in ASD. PMID- 21647151 TI - Genomic profiling in Down syndrome acute lymphoblastic leukemia identifies histone gene deletions associated with altered methylation profiles. AB - Patients with Down syndrome (DS) and acute lymphoblastic leukemia (ALL) have distinct clinical and biological features. Whereas most DS-ALL cases lack the sentinel cytogenetic lesions that guide risk assignment in childhood ALL, JAK2 mutations and CRLF2 overexpression are highly enriched. To further characterize the unique biology of DS-ALL, we performed genome-wide profiling of 58 DS-ALL and 68 non-DS (NDS) ALL cases by DNA copy number, loss of heterozygosity, gene expression and methylation analyses. We report a novel deletion within the 6p22 histone gene cluster as significantly more frequent in DS-ALL, occurring in 11 DS (22%) and only 2 NDS cases (3.1%) (Fisher's exact P=0.002). Homozygous deletions yielded significantly lower histone expression levels, and were associated with higher methylation levels, distinct spatial localization of methylated promoters and enrichment of highly methylated genes for specific pathways and transcription factor-binding motifs. Gene expression profiling demonstrated heterogeneity of DS ALL cases overall, with supervised analysis defining a 45-transcript signature associated with CRLF2 overexpression. Further characterization of pathways associated with histone deletions may identify opportunities for novel targeted interventions. PMID- 21647152 TI - Prevalence and prognostic value of IDH1 and IDH2 mutations in childhood AML: a study of the AML-BFM and DCOG study groups. AB - Mutations in the NADP(+)-dependent isocitrate dehydrogenase genes 1 and 2 (IDH1 and IDH2) have recently been found in adult acute myeloid leukemia (AML) patients with a prevalence rising up to 33%. To investigate the frequency of IDH1/2 mutations in pediatric AML, we characterized the mutational hotspot (exon 4) of these genes in diagnostic samples from 460 pediatric AML patients. Our analysis identified somatic IDH1/2 mutations in 4% of cases (IDH1 R132 n=8; IDH2 R140 n=10) and the minor allele of single-nucleotide polymorphism (SNP) rs11554137 in 47 children (10.2%). IDH mutations were associated with an intermediate age (P=0.008), FAB M1/M2 (P=0.013) and nucleophosmin1 mutations (P=0.001). In univariate analysis, IDH(mutated) compared with IDH(wildtype) patients showed a significantly improved overall survival (OS; P=0.032) but not event-free survival (EFS; P=0.14). However, multivariate analysis did not show independent prognostic significance. Children with at least one minor allele of IDH1 SNP rs11554137 had similar EFS (P=0.27) and OS (P=0.62) compared with major allele patients. Gene expression profiles of 12 IDH(mutated) were compared with 201 IDH(wildtype) patients to identify differentially expressed genes and pathways. Although only a small number of discriminating genes were identified, analysis revealed a deregulated tryptophan metabolism, and a significant downregulation of KYNU expression in IDH(mutated) cases. PMID- 21647153 TI - TAL1/SCL is downregulated upon histone deacetylase inhibition in T-cell acute lymphoblastic leukemia cells. AB - The transcription factor T-cell acute lymphocytic leukemia (TAL)-1 is a major T cell oncogene associated with poor prognosis in T-cell acute lymphoblastic leukemia (T-ALL). TAL1 binds histone deacetylase 1 and incubation with histone deacetylase inhibitors (HDACis) promotes apoptosis of leukemia cells obtained from TAL1 transgenic mice. Here, we show for the first time that TAL1 protein expression is strikingly downregulated upon histone deacetylase inhibition in T ALL cells. This is due to decreased TAL1 gene transcription in cells with native TAL1 promoter, and due to impaired TAL1 mRNA translation in cells that harbor the TAL1(d) microdeletion and consequently express TAL1 under the control of the SCL/TAL1 interrupting locus (SIL) promoter. Notably, HDACi-triggered apoptosis of T-ALL cells is significantly reversed by TAL1 forced overexpression. Our results indicate that the HDACi-mediated apoptotic program in T-ALL cells is partially dependent on their capacity to downregulate TAL1 and provide support for the therapeutic use of HDACi in T-ALL. PMID- 21647154 TI - IDH1 and IDH2 mutations in pediatric acute leukemia. AB - To investigate the frequency of isocitrate dehydrogenase 1 (IDH1) and 2 (IDH2) mutations in pediatric acute myeloid leukemia (AML) and acute lymphoid leukemia (ALL), we sequenced these genes in diagnostic samples from 515 patients (227 AMLs and 288 ALLs). Somatic IDH1/IDH2 mutations were rare in ALL (N=1), but were more common in AML, occurring in 3.5% (IDH1 N=3 and IDH2 N=5), with the frequency higher in AMLs with a normal karyotype (9.8%). The identified IDH1 mutations occurred in codon 132 resulting in replacement of arginine with either cysteine (N=3) or histidine (N=1). By contrast, mutations in IDH2 did not affect the homologous residue but instead altered codon 140, resulting in replacement of arginine with either glutamine (N=4) or tryptophan (N=1). Structural modeling of IDH2 suggested that codon 140 mutations disrupt the enzyme's ability to bind its substrate isocitrate. Accordingly, recombinant IDH2 R140Q/W were unable to carry out the decarboxylation of isocitrate to alpha-ketoglutarate (alpha-KG), but instead gained the neomorphic activity to reduce alpha-KG to R(-)-2 hydroxyglutarete (2-HG). Analysis of primary leukemic blasts confirmed high levels of 2-HG in AMLs with IDH1/IDH2 mutations. Interestingly, 3/5 AMLs with IDH2 mutations had FLT3-activating mutations, raising the possibility that these mutations cooperate in leukemogenesis. PMID- 21647155 TI - Donor KIR haplotype B improves progression-free and overall survival after allogeneic hematopoietic stem cell transplantation for multiple myeloma. PMID- 21647156 TI - Expansion of highly differentiated CD8+ T-cells or NK-cells in patients treated with dasatinib is associated with cytomegalovirus reactivation. AB - The tyrosine kinase inhibitor dasatinib exerts immunosuppressive effects on T cells and NK-cells in vitro. However, in some dasatinib-treated leukemia patients, clonal lymphocytosis with large granular lymphocyte (LGL) morphology develops, and this is associated with enhanced therapeutic responses. To elucidate the mechanistic basis for this paradoxical observation, we conducted detailed phenotypic and functional analyses of T-cell and NK-cell populations from 25 dasatinib-treated leukemia patients. All tested patients with LGL expansions (15/16) were cytomegalovirus (CMV) immunoglobulin (IgG) seropositive with high frequencies of CMV-specific CD8(+) T-cells; 5/16 LGL patients also experienced symptomatic CMV reactivation during dasatinib therapy. Expanded T cell and NK-cell populations exhibited late differentiated (CD27(-)CD57(+)) phenotypes; this was associated with a predisposition to apoptosis within the T cell compartment and impaired NK-cell cytotoxicity. Only 3/9 non-LGL patients were CMV IgG seropositive. Dasatinib inhibited in vitro lymphocyte functions, similarly in LGL patients and controls. Notably, distinct CD8(high) and CD8(low) T-cell subsets were observed in LGL patients; this phenotypic dichotomy was also apparent in CMV-specific CD8(+) T-cell populations, and exhibited features consistent with antigen-driven activation. In addition, plasma levels of IP-10, IL-6, monokine induced by interferon-gamma and interleukin-2R were significantly increased in LGL patients. These data provide evidence that dasatinib-associated LGL expansion is linked to CMV reactivation and suggest a potential mechanism for this phenomenon. PMID- 21647157 TI - Critical features of FAK-expressing AML bone marrow microenvironment through leukemia stem cell hijacking of mesenchymal stromal cells. PMID- 21647158 TI - Novel non-TCR chromosome translocations t(3;11)(q25;p13) and t(X;11)(q25;p13) activating LMO2 by juxtaposition with MBNL1 and STAG2. PMID- 21647160 TI - Intensified chemotherapy inspired by a pediatric regimen combined with allogeneic transplantation in adult patients with acute lymphoblastic leukemia up to the age of 40. AB - Event-free survival (EFS) at 5 years in pediatric acute lymphoblastic leukemia (ALL) is >80%. Outcome in adult ALL is still unsatisfactory, which is due to less cumulative dosing of chemotherapy and less strict adherence to timing of successive cycles. In the present phase II trial, we evaluated a pediatric regimen in adult patients with ALL under the age of 40. Treatment was according to the pediatric FRALLE approach for high-risk ALL patients and characterized by increased dosages of asparaginase, steroids, methotrexate and vincristin. However, allogeneic stem cell transplantation was offered to standard risk patients with a sibling donor and to all high-risk patients in contrast to the pediatric protocol. Feasibility was defined by achieving complete remission (CR) and completion of treatment within a strict timeframe in at least 60% of patients. In all, 54 patients were included with a median age of 26. CR was achieved in 49 patients (91%), of whom 33 completed treatment as scheduled (61%). Side effects primarily consisted of infections and occurred in 40% of patients. With a median follow-up of 32 months, EFS estimated 66% at 24 months and overall survival 72%. These data show that a dose-intensive pediatric regimen is feasible in adult ALL patients up to the age of 40. PMID- 21647159 TI - Notch signaling in mammalian hematopoietic stem cells. AB - Notch is a crucial cell signaling pathway in metazoan development. By means of cell-cell interactions, Notch signaling regulates cellular identity, proliferation, differentiation and apoptosis. Within the last decade, numerous studies have shown an important role for this pathway in the development and homeostasis of mammalian stem cell populations. Hematopoietic stem cells (HSCs) constitute a well-defined population that shows self-renewal and multi-lineage differentiation potential, with the clinically relevant capacity to repopulate the hematopoietic system of an adult organism. Here, we review the emergence, development and maintenance of HSCs during mammalian embryogenesis and adulthood, with respect to the role of Notch signaling in hematopoietic biology. PMID- 21647161 TI - Stable and reproducible engraftment of primary adult and pediatric acute myeloid leukemia in NSG mice. PMID- 21647162 TI - ZRF4, a combi-molecule with increased efficacy as compared with the individual components in chronic lymphocytic leukemia lymphocytes in vitro. PMID- 21647163 TI - The effects of exercise training on physical capacity, strength, body composition and functional performance among adults with spinal cord injury: a systematic review. AB - STUDY DESIGN: Systematic review. OBJECTIVES: To conduct a systematic review of evidence surrounding the effects of exercise on physical fitness in people with spinal cord injury (SCI). SETTING: Canada. METHODS: The review was limited to English-language studies (published prior to March 2010) of people with SCI that evaluated the effects of an exercise intervention on at least one of the four main components of physical fitness (physical capacity, muscular strength, body composition and functional performance). Studies reported at least one of the following outcomes: oxygen uptake/consumption, power output, peak work capacity, muscle strength, body composition, exercise performance or functional performance. A total of 166 studies were identified. After screening, 82 studies (69 chronic SCI; 13 acute SCI) were included in the review. The quality of evidence derived from each study was evaluated using established procedures. RESULTS: Most studies were of low quality; however, the evidence was consistent that exercise is effective in improving aspects of fitness. There is strong evidence that exercise, performed 2-3 times per week at moderate-to-vigorous intensity, increases physical capacity and muscular strength in the chronic SCI population; the evidence is not strong with respect to the effects of exercise on body composition or functional performance. There were insufficient high-quality studies in the acute SCI population to draw any conclusions. CONCLUSIONS: In the chronic SCI population, there is good evidence that exercise is effective in improving both physical capacity and muscular strength, but insufficient quality evidence to draw meaningful conclusions on its effect on body composition or functional capacity. PMID- 21647164 TI - The development of evidence-informed physical activity guidelines for adults with spinal cord injury. AB - OBJECTIVES: To systematically develop evidence-informed physical activity guidelines to improve physical fitness in people with spinal cord injury (SCI). SETTING: This study was conducted in Canada. METHODS: The Appraisal of Guidelines, Research and Evaluation II guideline development protocol was used to develop exercise guidelines to improve physical capacity and muscular strength. The evidence base for the guideline development process consisted of a systematic review and quality appraisal of research examining the effects of exercise on physical fitness among people with SCI. A multidisciplinary expert panel deliberated the evidence and generated the guidelines. Pilot testing led to refinement of the wording and presentation of the guidelines. RESULTS: The expert panel generated the following guidelines: for important fitness benefits, adults with a SCI should engage in (a) at least 20 min of moderate to vigorous intensity aerobic activity two times per week and (b) strength training exercises two times per week, consisting of three sets of 8-10 repetitions of each exercise for each major muscle group. CONCLUSION: People with SCI, clinicians, researchers and fitness programmers are encouraged to adopt these rigorously developed guidelines. PMID- 21647165 TI - Is the outcome in acute spinal cord ischaemia different from that in traumatic spinal cord injury? A cross-sectional analysis of the neurological and functional outcome in a cohort of 93 paraplegics. PMID- 21647166 TI - Evaluation of a task-oriented client-centered upper extremity skilled performance training module in persons with tetraplegia. AB - STUDY DESIGN: Clinical intervention study. OBJECTIVE: Arm hand skilled performance (AHSP) has a major role in the rehabilitation of persons with cervical spinal cord injury (C-SCI). A task-oriented client-centered upper extremity skilled performance-training (ToCUEST) module was developed. The present study aims to evaluate effects of ToCUEST on specific and general AHSP in C-SCI, during and after rehabilitation. SETTING: Adelante Rehabilitation Centre (The Netherlands). METHODS: C-SCI persons, either during or after rehabilitation, completed the ToCUEST training; a control group of patients during rehabilitation received standard rehabilitation. The training module was focused on patient's individual needs, combining principles of motor learning and training physiology. Three self-selected goals were trained 3 days per week, in three sessions of 30 min per day for 8 weeks. Measures were taken at 3 months after the start of the active rehabilitation (T0), before training (T1), after training (T2), at follow up (3 months post-training; T3) and at discharge. RESULTS: For both ToCUEST groups during (n=11) and after (n=12) rehabilitation, an improvement (P<0.001) on specific AHSP (using the Goal Attainment Scale and the Canadian Occupational Performance Measure (satisfaction and performance)) was found between T1-T2 and T1-T3. Also an improvement (P<0.02) in general AHSP (using the Van Lieshout Test, the QIF and the motor FIM) was found in the total ToCUEST group between T1-T2 and T1-T3. No significant difference in improvement of general AHSP was found between the ToCUEST rehab and control group. CONCLUSION: The ToCUEST module leads to improvement in AHSP, not only persons with C-SCI during rehabilitation, but also after finishing rehabilitation. These effects remain at follow-up. PMID- 21647167 TI - Myxopapillary ependymoma: correlation of clinical and imaging features with surgical resectability in a series with long-term follow-up. AB - STUDY DESIGN: Retrospective case series. OBJECTIVES: The objective of this study is to identify imaging and intraoperative characteristics that may predict surgical resection for myxopapillary ependymoma (MPE). The diffuse involvement in the conus-filum region makes complete resection challenging. The preoperative characteristics that may estimate the extent of resection has not been reported. SETTING: Toronto, Canada. METHODS: All MPE cases between 1972 and 2005 at a single institution were identified and reexamined by a neuropathologist. Neurological outcomes (Frankel scale), clinical features, operative findings, pre and postoperative imaging results were reviewed. RESULTS: A total of 18 operations were performed on 15 MPE patients (8 females/7 males; age range: 18-71 years). Median postoperative follow-up was 56 months. Three patients (17%) developed tumor regrowth requiring reoperations. Preoperative magnetic resonance imaging (MRI; in 14/18 procedures) determined that tumors involved the conus in 70% of cases, which was significantly associated with intraoperative findings (P=0.02). Complete microsurgical resection was accomplished in 4 out of 7 cases where conus was not involved, but in only 1 out of 10 cases with conus involvement (P=0.056). The degree of conus involvement in one case was unclear. None of patients with total surgical resection developed recurrence. All patients survived at long-term follow-up. CONCLUSION: Our series is the first to correlate MPE involvement to conus medullaris on preoperative MRI with intraoperative findings, and examine its significance on surgical resectability. This information could guide clinicians in preoperative planning and advising patients on treatment options and potential risks/benefits. MRI is very sensitive (100%) and moderately specific (67%) in detecting direct anatomical contact between conus and MPE tumors. PMID- 21647168 TI - Priapism in acute spinal cord injury. AB - STUDY DESIGN: Prospective literature review; search via Oldmedline (1958-1965), Medline (1966-2005) and Pubmed. OBJECTIVES: The objective of this study is to review the pathophysiology of priapism in acute traumatic spinal cord injury (SCI); to determine the incidence of priapism in traumatic SCI, whether or not priapism is associated with incomplete or only complete SCI and whether and what treatment might be required. METHODS: This is a review article based upon the available literature in this area. RESULTS AND CONCLUSIONS: Priapism that follows acute traumatic SCI is high-flow (non-ischaemic) priapism, that is, the blood within the corpus is arterial in nature. Priapism does not occur in all patients with acute SCI. The literature does not allow us to determine in what proportion of patients priapism occurs. Priapism has been reported following a wide variety of spinal cord pathologies including acute SCI, transverse myelitis and postoperative extradural haematoma. In all patients, priapism is associated with complete motor and sensory (American Spinal Injury Association A) paraplegia. Priapism has been reported following spinal shock. Following traumatic SCI, priapism usually settles rapidly without specific treatment being required. Priapism occurs at the moment of complete motor and sensory paraplegia, it does not occur following a delay. There are medicolegal implications: the presence/absence of priapism assists in determining when the complete spinal cord lesion occurred. PMID- 21647169 TI - Modulation of soluble and particulate antigen transport in afferent lymph by monophosphoryl lipid A. AB - Vaccine adjuvants stimulate the innate immune system and determine the outcome of the immune response induced. A better understanding of their action is therefore crucial to the development of new and safer vaccines. Monophosphoryl lipid A (MPL), a 'detoxified' version of lipolysaccharide, is a promising new adjuvant component in human vaccines. The present study uses an ovine lymphatic cannulation model to study cell recruitment and antigen transport from the injection site into the afferent lymph, and how this is modulated by co-injection with MPL. Compared with saline, MPL injections caused only minor variations in lymph flow and no difference in cell number migrating into the lymph. MPL did, however, cause a significantly increased recruitment of neutrophils and monocytes, but not dendritic cells (DC) into the lymph for the first 12 h. Soluble ovalbumin (OVA) antigen flowed freely into the lymph over a 24-h period and was slightly reduced at 6-9 h in the MPL-injected sites. OVA-coated fluorescent 1-MU beads were initially transported predominantly by neutrophils and, from 24 to 72 h, by DC. MPL induced an increased and more sustained transport of beads by neutrophils and monocytes although it did not increase the phagocytic capacity of these cells. In contrast to aluminium adjuvant, MPL did not increase bead transport by DC at the later time point. These studies provide important new insights in the in vivo action of different adjuvants and the initial events that set up an immune response after vaccination. PMID- 21647170 TI - Seasoned adaptive antibody immunity for highly pathogenic pandemic influenza in humans. AB - Fundamentally new approaches are required for the development of vaccines to pre empt and protect against emerging and pandemic influenzas. Current strategies involve post-emergent homotypic vaccines that are modelled upon select circulating 'seasonal' influenzas, but cannot induce cross-strain protection against newly evolved or zoonotically introduced highly pathogenic influenza (HPI). Avian H5N1 and the less-lethal 2009 H1N1 and their reassortants loom as candidates to seed a future HPI pandemic. Therefore, more universal 'seasoned' vaccine approaches are urgently needed for heterotypic protection ahead of time. Pivotal to this is the need to understand mechanisms that can deliver broad strain protection. Heterotypic and heterosubtypic humoral immunities have largely been overlooked for influenza cross-protection, with most 'seasoned' vaccine efforts for humans focussed on heterotypic cellular immunity. However, 5 years ago we began to identify direct and indirect indicators of humoral-herd immunity to protein sites preserved among H1N1, H3N2 and H5N1 influenzas. Since then the evidence for cross-protective antibodies in humans has been accumulating. Now proposed is a rationale to stimulate and enhance pre-existing heterotypic humoral responses that, together with cell-mediated initiatives, will deliver pre-emptive and universal human protection against emerging epidemic and pandemic influenzas. PMID- 21647171 TI - CD4 T cells play important roles in maintaining IL-17-producing gammadelta T-cell subsets in naive animals. AB - A proportional balance between alphabeta and gammadelta T-cell subsets in the periphery is exceedingly well maintained by a homeostatic mechanism. However, a cellular mechanism underlying the regulation remains undefined. We recently reported that a subset of developing gammadelta T cells spontaneously acquires interleukin (IL)-17-producing capacity even within naive animals through a transforming growth factor (TGF)beta1-dependent mechanism, thus considered 'innate' IL-17-producing cells. Here, we report that gammadelta T cells generated within alphabeta T cell (or CD4 T cell)-deficient environments displayed altered cytokine profiles; particularly, 'innate' IL-17 expression was significantly impaired compared with those in wild-type mice. Impaired IL-17 production in gammadelta T cells was directly related to CD4 T-cell deficiency, because depletion of CD4 T cells in wild-type mice diminished and adoptive CD4 T-cell transfer into T-cell receptor beta-/- mice restored IL-17 expression in gammadelta T cells. CD4 T cell-mediated IL-17 expression required TGFbeta1. Moreover, Th17 but not Th1 or Th2 effector CD4 T cells were highly efficient in enhancing gammadelta T-cell IL-17 expression. Taken together, our results highlight a novel CD4 T cell-dependent mechanism that shapes the generation of IL 17+ gammadelta T cells in naive settings. PMID- 21647172 TI - SAP suppresses the development of experimental autoimmune encephalomyelitis in C57BL/6 mice. AB - Experimental autoimmune encephalomyelitis (EAE) is a CD4(+) T cell-mediated disease of the central nervous system. Serum amyloid P component (SAP) is a highly conserved plasma protein named for its universal presence in amyloid deposits. Here we report that SAP-transgenic mice had unexpectedly attenuated EAE due to impaired encephalitogenic responses. Following induction with myelin oligodendroglial glycoprotein (MOG) peptide 35-55 in complete Freund's adjuvant, SAP-transgenic mice showed reduced spinal cord inflammation with lower severity of EAE attacks as compared with control C57BL/6 mice. However, in SAP-Knockout mice, the severity of EAE is enhanced. Adoptive transfer of Ag-restimulated T cells from wild type to SAP-transgenic mice, or transfer of SAP-transgenic Ag restimulated T cells to control mice, induced milder EAE. T cells from MOG-primed SAP-transgenic mice showed weak proliferative responses. Furthermore, in SAP transgenic mice, there is little infiltration of CD45-positive cells in the spinal cord. In vitro, SAP suppressed the secretion of interleukin-2 stimulated by P-selectin and blocked P-selectin binding to T cells. Moreover, SAP could change the affinity between alpha4-integrin and T cells. These data suggested that SAP could antagonize the development of the acute phase of inflammation accompanying EAE by modulating the function of P-selectin. PMID- 21647173 TI - Toll-like receptor 2 siRNA suppresses corneal inflammation and attenuates Aspergillus fumigatus keratitis in rats. AB - Toll-like receptors (TLRs) are key components of innate immunity that detect microbial infection and trigger host defense responses. However, they are capable of initiating both protective and damaging immune responses, as exaggerated expression of inflammatory components can have devastating effects on the host. We previously reported that TLR2 in corneal epithelium has an important role in the pathogenesis of fungal keratitis, however, how the corneal inflammation is modulated remains to be elucidated. This study aims to investigate the effect of targeting TLR2 on Aspergillus fumigatus keratitis in rats. The control or TLR2 small interfering RNA (siRNA) was applied sub-conjunctively and topically to the cornea. TLR2 immunostaining was performed to determine the feasibility of TLR2 siRNA delivery. Production of inflammatory cytokines and chemokines were determined by real-time quantitative PCR. Polymorphonuclear leukocyte (PMN) infiltration was assessed by myeloperoxidase activity. It was found that rat corneas treated with TLR2 siRNA showed a significant reduction of TLR2 expression in corneal epithelium. TLR2 siRNA treatment improved the outcome of keratitis, which was characterized by decreased corneal opacity, less corneal perforation, suppressed PMN infiltration, reduced production of inflammatory cytokines and chemokines, and less fungal burden. In conclusion, TLR2 siRNA treatment attenuated A. fumigatus keratitis by suppressing corneal inflammation and preventing fungal invasion, suggesting a novel avenue to control fungal infection and avert damage caused by excessive inflammation. PMID- 21647174 TI - Berberine and evodiamine influence serotonin transporter (5-HTT) expression via the 5-HTT-linked polymorphic region. AB - The aim of this study was to investigate the effect of berberine and evodiamine on serotonin transporter (5-HTT) expression and then test how allelic variations previously identified in the promoter region could modulate that effect in the serotonergic neuronal cell line RN46A. Both berberine and evodiamine, alone and in combination, increased 5-HTT mRNA and protein expression significantly across the various alleles. When tested against the S, XS(11), L(G), L(A), XL(17), and XL(18) alleles, respectively, 100 MUM berberine increased 5-HTT promoter activities by 67%, 128.7%, 106.9%, 100.4%, 26.2% and 82%, 2 MUM evodiamine increased 5-HTT promoter activities by 216.7%, 81.6%, 305.6%, 181.5%, 175.3% and 102.2%. Berberine and evodiamine increased 5-HTT promoter activity differently depending on the genetic variation of the 5-HTTLPR polymorphism. This study has provided a convincing example of how herbal compounds influence the expression of one of the most intensively studied psychiatric candidate genes, the serotonin transporter. PMID- 21647175 TI - Gene expression in blood is associated with risperidone response in children with autism spectrum disorders. AB - Children with autism spectrum disorders (ASDs) often have severe behavioral problems. Not all children with these problems respond to atypical antipsychotic medications; therefore, we investigated whether peripheral blood gene expression before treatment with risperidone, an atypical antipsychotic, was associated with improvements in severe behavioral disturbances 8 weeks following risperidone treatment in 42 ASD subjects (age 112.7+/-51.2 months). Exon expression levels in blood before risperidone treatment were compared with pre-post risperidone change in Aberrant Behavior Checklist-Irritability (ABC-I) scores. Expression of exons within five genes was correlated with change in ABC-I scores across all risperidone-treated subjects: GBP6, RABL5, RNF213, NFKBID and RNF40 (alpha<0.001). RNF40 is located at 16p11.2, a region implicated in autism and schizophrenia. Thus, these genes expressed before treatment were associated with subsequent clinical response. Future studies will be needed to confirm these results and determine whether this expression profile is associated with risperidone response in other disorders, or alternative antipsychotic response within ASD. PMID- 21647176 TI - XRCC1 Arg399Gln and Arg194Trp polymorphisms in prostate cancer risk: a meta analysis. AB - Epidemiological studies have evaluated the association between X-ray repair cross complementing group 1 gene (XRCC1) Arg399Gln and Arg194Trp polymorphisms and risk of prostate cancer (PCa). However, the results from the published studies on the association between these two XRCC1 polymorphisms and PCa risk are conflicting. To derive a more precise estimation of association between the XRCC1 polymorphisms and risk of PCa, we performed a meta-analysis. A comprehensive search was conducted to identify all case-control studies of XRCC1 polymorphisms and PCa risk. We used odds ratios (ORs) with 95% confidence intervals (CIs) to assess the strength of the association. Overall, we found that both Arg399Gln and Arg194Trp polymorphisms were not significantly associated with PCa risk. However, in stratified analysis by ethnicity, we found that the Arg399Gln polymorphism was significantly associated with PCa risk in Asian population (Gln/Gln vs Arg/Arg: OR=1.46, 95% CI: 1.05-2.03, P=0.03; Gln/Gln vs Arg/Gln+Arg/Arg: OR=1.48, 95% CI: 1.12-1.95, P=0.01). In this meta-analysis, we found that both Arg399Gln and Arg194Trp polymorphisms were not related to overall PCa risk. However, in subgroup analysis we found a suggestion that XRCC1 399Gln allele might be a low penetrent risk factor for PCa only in Asian men. PMID- 21647177 TI - To go or not to go: the "itchy" effect on the destiny of hematopoietic stem cells. PMID- 21647179 TI - Therapy: antisense inhibitor of apo B has potent lipoprotein effects. PMID- 21647180 TI - Surgery: predicting acute type B aortic dissection-size does not matter. PMID- 21647181 TI - Venous thromboembolism: optimal therapy duration assessed. PMID- 21647182 TI - Atrial fibrillation: new-onset AF in women--an independent risk factor or a 'canary in a coal mine'? PMID- 21647183 TI - Electrical nerve stimulation for overactive bladder in children. AB - Overactive bladder (OAB) is a urological condition that is frequently observed in children and requires treatment. Standard urotherapy is usually the first line of treatment; however, children with severe OAB do not respond to this therapy. Antimuscarinic drugs may be used, but complete resolution of symptoms with this medication is low. Studies, including two randomized clinical trials, have demonstrated that electrical nerve stimulation (ENS) can be used successfully for OAB in children. Electrodes have been placed in the genitals, anus, and intravesical area, and on the tibial posterior nerve, as well as used transcutaneously or implanted during a surgical procedure. This Review will discuss the use of ENS in children with OAB, with particular focus on the putative mechanisms of action, the different ENS techniques available, and clinical results for this therapy. PMID- 21647184 TI - Pediatrics: Repeated botulinum toxin-A injections safe in children. PMID- 21647185 TI - Urinary tract obstruction: Renal function reserve predicts long-term renal deterioration in children with posterior urethral valve. PMID- 21647187 TI - Thyroid gland: TSH mediates increased risk of PTC in Hashimoto thyroiditis. PMID- 21647188 TI - Reproductive endocrinology: Benefits of metformin in PCOS. PMID- 21647189 TI - GLP-1 and energy balance: an integrated model of short-term and long-term control. AB - Glucagon-like peptide 1 (GLP-1), a peptide secreted from the intestine in response to nutrient ingestion, is perhaps best known for its effect on glucose stimulated insulin secretion. GLP-1 is also secreted from neurons in the caudal brainstem, and it is well-established that, in rodents, central administration of GLP-1 potently reduces food intake. Over the past decade, GLP-1 has emerged not only as an essential component of the system that regulates blood glucose levels but also as a viable therapeutic target for the treatment of type 2 diabetes mellitus. However, although GLP-1 receptor agonists are known to produce modest but statistically significant weight loss in patients with diabetes mellitus, our knowledge of how endogenous GLP-1 regulates food intake and body weight remains limited. The purpose of this Review is to discuss the evolution of our understanding of how endogenous GLP-1 modulates energy balance. Specifically, we consider contributions of both central and peripheral GLP-1 and propose an integrated model of short-term and long-term control of energy balance. Finally, we discuss this model with respect to current GLP-1-based therapies and suggest ongoing research in order to maximize the effectiveness of GLP-1-based treatment of obesity. PMID- 21647190 TI - Genetics: the crystal ball clears for breast cancer therapy? PMID- 21647191 TI - Treatment modalities: HIFU is effective for unresectable HCC. PMID- 21647192 TI - Chemotherapy: Perioperative therapy improves gastroesophageal cancer survival. PMID- 21647193 TI - Local treatment of metastatic cancer--killing the seed or disturbing the soil? AB - The fact that local therapy of the primary tumor is futile in the presence of metastatic disease is almost considered axiomatic among oncologists. However, this perception is now being challenged by new laboratory and clinical data. Results from animal models have demonstrated that some primary tumors release factors that enter the circulation which, by mobilizing cells from bone marrow, render distant organs more receptive to metastasis. Clinical observations in renal, breast, and prostate cancer are all consistent with the hypothesis that treatment directed against the primary tumor might retard progression of existing metastases. This hypothesis is amenable to testing by randomized trials of local therapy to the primary site in patients with metastatic cancer. PMID- 21647194 TI - Health services: How can we address cancer care after a natural disaster? PMID- 21647196 TI - Targeted therapies: Radiopeptide therapy improves outcomes for neuroendocrine cancers. PMID- 21647195 TI - MicroRNAs in body fluids--the mix of hormones and biomarkers. AB - Since the discovery of microRNAs (miRNAs), the study of these small noncoding RNAs has steadily increased and more than 10,000 papers have already been published. The great interest in miRNAs reflects their central role in gene expression regulation and the implication of miRNA-specific aberrant expression in the pathogenesis of cancer, cardiac, immune-related and other diseases. Another avenue of current research is the study of circulating miRNAs in serum, plasma, and other body fluids--miRNAs may act not only within cells, but also at other sites within the body. The presence of miRNAs in body fluids may represent a gold mine of noninvasive biomarkers in cancer. Since deregulated miRNA expression is an early event in tumorigenesis, measuring circulating miRNA levels may also be useful for early cancer detection, which can contribute greatly to the success of treatment. In this Review, we discuss the role of fluid-expressed miRNAs as reliable cancer biomarkers and treatment-response predictors as well as potential new patient selection criteria for clinical trials. In addition, we explore the concept that miRNAs could function as hormones. PMID- 21647197 TI - Surgical oncology: Good things don't come to those who watchfully wait. PMID- 21647198 TI - Gastrointestinal effects of aspirin. AB - Aspirin is being used as an effective analgesic and anti-inflammatory agent at doses >325 mg daily. At low doses (75-325 mg daily), aspirin is the key antiplatelet drug in the pharmacological prevention of cardiovascular diseases. Topical and systemic effects of aspirin in the gastrointestinal mucosa are associated with mucosal damage in the upper and lower gastrointestinal tract. The risk of upper gastrointestinal bleeding with aspirin is increased with old age, male sex, ulcer history and concomitant medication with NSAIDs, cyclooxygenase 2 selective inhibitors, corticosteroids or other antithrombotic agents. In some patients, the cardiovascular benefits of low-dose aspirin might be overcome by the risk of gastrointestinal complications, but withdrawal of aspirin therapy can precipitate a cardiovascular event. These patients will need concomitant therapy with antisecretory agents, especially PPIs, to reduce the gastrointestinal risk. Eradication of Helicobacter pylori infection might be an additional option in patients with a history of ulcer. Furthermore, there is growing evidence that long-term use of aspirin decreases the risk of colorectal cancer, even at low doses. As aspirin is one of the most prescribed drugs worldwide and its clinical impact is huge, physicians need to consider the benefits and harms for each individual patient in order to maximize the benefits of aspirin. PMID- 21647199 TI - Anti-HER agents in gastric cancer: from bench to bedside. AB - Despite some advances in the past few years, the search for effective treatment modalities for advanced gastric and gastro-esophageal junction cancer is far from over. Available data clearly demonstrate that the development of new drugs will have little, if any, chance of success if it is not guided by in-depth knowledge of disease biology. However, using biologic agents to target key molecular pathways, such as those regulated by human epidermal growth factor receptor (HER) family members, may be effective. Indeed, the positive results achieved by the anti-HER2 agent trastuzumab in a phase III trial in HER2-positive patients support this approach. Many new anti-HER molecules are now under evaluation for the treatment of gastric and gastro-esophageal junction cancer, but so far attempts to identify reliable predictive factors from phase I and II trials have produced inconclusive results. In addition, large phase III trials are still being conducted in molecularly unselected populations. Refining patient selection is essential to maximize the benefit of targeted agents, to avoid significant toxicities and for the development of alternative therapeutic approaches in patients who have nonresponsive disease. PMID- 21647201 TI - Epilepsy: HLA alleles linked to carbamazepine hypersensitivity. PMID- 21647200 TI - Pathogenesis and biomarkers of carcinogenesis in ulcerative colitis. AB - One of the most serious complications of ulcerative colitis is the development of colorectal cancer. Screening patients with ulcerative colitis by standard histological examination of random intestinal biopsy samples might be inefficient as a method of cancer surveillance. This Review focuses on the current understanding of the pathogenesis of ulcerative colitis-associated colorectal cancer and how this knowledge can be transferred into patient management to assist clinicians and pathologists in identifying patients with ulcerative colitis who have an increased risk of colorectal cancer. Inflammation-driven mechanisms of DNA damage, including the generation and effects of reactive oxygen species, microsatellite instability, telomere shortening and chromosomal instability, are reviewed, as are the molecular responses to genomic stress. We also discuss how these mechanisms can be translated into usable biomarkers. Although progress has been made in the understanding of inflammation-driven carcinogenesis, markers based on these findings possess insufficient sensitivity or specificity to be usable as reliable biomarkers for risk of colorectal cancer development in patients with ulcerative colitis. However, screening for mutations in p53 could be relevant in the surveillance of patients with ulcerative colitis. Several other new biomarkers, including senescence markers and alpha-methylacyl CoA-racemase, might be future candidates for preneoplastic markers in ulcerative colitis. PMID- 21647202 TI - Emerging therapeutic targets in schwannomas and other merlin-deficient tumors. AB - Deficiency of the tumor suppressor protein merlin leads to the development of benign tumors of the nervous system such as schwannomas, ependymomas and meningiomas. These tumors can occur spontaneously or as part of a tumor predisposition syndrome called neurofibromatosis type 2 (NF2), which involves multiple tumors. Schwannomas are the hallmark tumors of NF2 and are the most frequent and well-characterized of the merlin-deficient tumors. Surgery or radiotherapy are used to treat single tumors and can leave the patient with substantial morbidity. Limitations of other treatment options for merlin deficient tumors, such as the lack of effectiveness of chemotherapy, have led to an urgent requirement for new pharmaceutical therapies. Merlin-deficient tumors are genetically well-defined, which allows rational testing of new molecular therapies that have been developed and successfully used to treat various cancers in the past few years. This Review centers on four key families of receptor tyrosine kinases-the ErbB family, platelet-derived growth factor receptor beta, insulin-like growth factor 1 receptor, and vascular endothelial growth factor receptors-focusing on their role in schwannoma pathobiology and the therapeutic potential of targeting these receptors and their downstream signaling pathways. PMID- 21647203 TI - Anti-CCP antibodies: the past, the present and the future. AB - Rheumatoid arthritis (RA) is an autoimmune disease characterized by autoantibodies against citrullinated antigens. The importance of citrulline for the epitopes bound by these autoantibodies, referred to as ACPA (anti citrullinated peptide/protein antibodies), was first described in 1998. In addition to citrullinated proteins, cyclic citrullinated peptides (CCP) can also be used as test substrates for detecting ACPA. The standard test for these antibodies is the second-generation CCP (CCP2) test, which is one of the best in terms of sensitivity and specificity. The generation of ACPA is an early event in the disease course, and is dependent on the presence of certain MHC class II alleles. ACPA in the inflamed synovium have been shown to associate with citrullinated antigens to form immune complexes, resulting in progression of the inflammatory process. The involvement of ACPA in the chronicity of RA is probably the reason why ACPA-positive patients have a more erosive disease course than ACPA-negative patients. The presence of ACPA has been included in the 2010 RA classification criteria. Thus, it is important to further standardize ACPA testing, for example by including an internal serum standard, which may lead to a better distinction between low and high ACPA levels. PMID- 21647205 TI - Connective tissue diseases: Swallowing tolerance in a tablet might be effective treatment strategy for SLE. PMID- 21647204 TI - Pathogenesis of systemic juvenile idiopathic arthritis: some answers, more questions. AB - Systemic juvenile idiopathic arthritis (sJIA) has long been recognized as unique among childhood arthritides, because of its distinctive clinical and epidemiological features, including an association with macrophage activation syndrome. Here, we summarize research into sJIA pathogenesis. The triggers of disease are unknown, although infections are suspects. Once initiated, sJIA seems to be driven by innate proinflammatory cytokines. Endogenous Toll-like receptor ligands, including S100 proteins, probably synergize with cytokines to perpetuate inflammation. These and other findings support the hypothesis that sJIA is an autoinflammatory condition. Indeed, IL-1 is implicated as a pivotal cytokine, but the source of excess IL-1 activity remains obscure and the role of IL-1 in chronic arthritis is less clear. Another hypothesis is that a form of hemophagocytic lymphohistiocytosis underlies sJIA, with varying degrees of its expression across the spectrum of disease. Alternatively, sJIA with MAS might be a genetically distinct subtype. Yet another hypothesis proposes that inadequate downregulation of immune activation is central to sJIA, supporting evidence for which includes 'alternative activation' of monocyte and macrophages and possible deficiencies in IL-10 and T regulatory cells. Some altered immune phenotypes persist during clinically inactive disease, which suggests that this stage might represent compensated inflammation. Despite much progress being made, many questions remain, providing fertile ground for future research. PMID- 21647206 TI - Characterization of episodes of irritable bowel syndrome using ecological momentary assessment. AB - OBJECTIVES: Patients with irritable bowel syndrome (IBS) report that symptoms occur as episodes. The nature and frequency of episodes have not been well studied. METHODS: Using modified ecological momentary assessment (EMA), we examined clinical factors attributed to IBS symptom episodes and compared them with nonsymptom episode periods in patients with IBS-D (N=21), IBS-C (N=18), or IBS-M (N=19), and healthy controls (N=19). Symptoms were rated over 14 days on a visual ordinal scale (VOS: 0-10) randomly in morning, midday, and evening, and at wake up, bedtime, prebowel movement, and postbowel movement. Scores were evaluated for total group and across subgroups and between EMA and daily diary cards on the same day. RESULTS: Subjects (n=57/59) reported symptom episodes 34% of the time. Episodes showed significantly higher pain levels (3.6 vs. 1.64, P<0.0001), bloating (4.57 vs. 3.02, P<0.0001), stress (3.54 vs. 2.59, P<0.0001), and decreased well-being (5.29 vs. 6.16, P<0.0001). Episode frequency/2 weeks was greatest for IBS-D (10.7+/-7.05) than IBS-C (8.4+/-5.76) and IBS-M (7.1+/-4.45) (P=nonsignificant). IBS-D also had shorter episodes (9 h 23 min) compared with IBS-M (15 h 01 min) and IBS-C (15 h 25 min) (P<0.04). Stool frequency and looser consistency were greater with IBS-D and similar between IBS-C and IBS-M. Abdominal pain was the greatest predictor of episode status. Diary card ratings of pain and stool frequency overestimate levels reported by EMA. CONCLUSIONS: Episodes of IBS are associated with greater pain (strongest relationship), bloating, and stress scores, and poorer global well-being. Compared with IBS-D, IBS-C and IBS-M are similar in clinical features. Patients overreport pain and stool frequency by diary compared with EMA. PMID- 21647207 TI - Mucin expression pattern in pancreatic diseases: findings from EUS-guided fine needle aspiration biopsies. AB - OBJECTIVES: Alterations in mucin (MUC) glycosylation and expression have been described in cancer. Endoscopic ultrasound-guided fine-needle aspiration (EUS FNA) can provide material for molecular biology analysis. This study assessed the feasibility of evaluating MUC expression from material obtained by EUS-FNA and studied the profile of MUC expression in benign and malignant pancreatic lesions. METHODS: A total of 90 patients with solid or cystic pancreatic lesions underwent FNA. The aspirated material was used for cytological analysis and RNA extraction to assess the expression pattern of MUCs by reverse transcription-PCR with primers specific for the MUC1, MUC2, MUC3, MUC4, MUC5A, MUC5B, MUC6, and MUC7 genes. RESULTS: RNA extraction was successful in 81% of the biopsies. The prevalences of MUC1, MUC2, MUC4, and MUC7 in ductal adenocarcinoma were 57.7, 51.4, 18.9, and 73.0%, respectively. Fifty percent of benign lesions and neuroendocrine tumors (NETs), and 63% of intraductal papillary mucinous neoplasms (IPMNs) were positive for MUC1. Twenty-five percent of benign lesions, 86% of NETs, and 47% of IPMNs were positive for MUC2. Of NETs, 50% were positive for MUC1, and 14% were positive for MUC7. None of the benign lesions or NETs expressed MUC4. MUC7 expression was highly significant for adenocarcinoma (P=0.007) and borderline for IPMN (P=0.05). MUC7 was expressed in 37.5% of chronic pancreatitis cases. CONCLUSIONS: RNA can be extracted from samples obtained under EUS-FNA. MUC7 could serve as a potential biological marker to identify malignant lesions, especially pancreatic adenocarcinoma. PMID- 21647208 TI - Persistency of priors-induced bias in decision behavior and the FMRI signal. AB - It is well known that people take advantage of prior knowledge to bias decisions. To investigate this phenomenon behaviorally and in the brain, we acquired fMRI data while human subjects viewed ambiguous abstract shapes and decided whether a shape was of Category A (smoother) or B (bumpier). The decision was made in the context of one of two prior knowledge cues, 80/20 and 50/50. The 80/20 cue indicated that upcoming shapes had an 80% probability of being of one category, e.g., B, and a 20% probability of being of the other. The 50/50 cue indicated that upcoming shapes had an equal probability of being of either category. The ideal observer would bias decisions in favor of the indicated alternative at 80/20 and show zero bias at 50/50. We found that subjects did bias their decisions in the predicted direction at 80/20 but did not show zero bias at 50/50. Instead, at 50/50 the subjects retained biases of the same sign as their 80/20 biases, though of diminished magnitude. The signature of a persistent though diminished bias at 50/50 was also evident in fMRI data from frontal and parietal regions previously implicated in decision-making. As a control, we acquired fMRI data from naive subjects who experienced only the 50/50 stimulus distributions during both the pre-scan training and the fMRI experiment. The behavioral and fMRI data from the naive subjects reflected decision biases closer to those of the ideal observer than those of the prior knowledge subjects at 50/50. The results indicate that practice making decisions in the context of non equal prior probabilities biases decisions made later when prior probabilities are equal. This finding may be related to the "anchoring and adjustment" strategy described in the psychology, economics, and marketing literatures, in which subjects adjust a first approximation response - the "anchor" - based on additional information, typically applying insufficient adjustment relative to the ideal observer. PMID- 21647209 TI - HumMod: A Modeling Environment for the Simulation of Integrative Human Physiology. AB - Mathematical models and simulations are important tools in discovering key causal relationships governing physiological processes. Simulations guide and improve outcomes of medical interventions involving complex physiology. We developed HumMod, a Windows-based model of integrative human physiology. HumMod consists of 5000 variables describing cardiovascular, respiratory, renal, neural, endocrine, skeletal muscle, and metabolic physiology. The model is constructed from empirical data obtained from peer-reviewed physiological literature. All model details, including variables, parameters, and quantitative relationships, are described in Extensible Markup Language (XML) files. The executable (HumMod.exe) parses the XML and displays the results of the physiological simulations. The XML description of physiology in HumMod's modeling environment allows investigators to add detailed descriptions of human physiology to test new concepts. Additional or revised XML content is parsed and incorporated into the model. The model accurately predicts both qualitative and quantitative changes in clinical and experimental responses. The model is useful in understanding proposed physiological mechanisms and physiological interactions that are not evident, allowing one to observe higher level emergent properties of the complex physiological systems. HumMod has many uses, for instance, analysis of renal control of blood pressure, central role of the liver in creating and maintaining insulin resistance, and mechanisms causing orthostatic hypotension in astronauts. Users simulate different physiological and pathophysiological situations by interactively altering numerical parameters and viewing time-dependent responses. HumMod provides a modeling environment to understand the complex interactions of integrative physiology. HumMod can be downloaded at http://hummod.org. PMID- 21647210 TI - Gelsemine and Gelsemium sempervirens L. Extracts in Animal Behavioral Test: Comments and Related Biases. PMID- 21647212 TI - The evolution of the brain, the human nature of cortical circuits, and intellectual creativity. AB - The tremendous expansion and the differentiation of the neocortex constitute two major events in the evolution of the mammalian brain. The increase in size and complexity of our brains opened the way to a spectacular development of cognitive and mental skills. This expansion during evolution facilitated the addition of microcircuits with a similar basic structure, which increased the complexity of the human brain and contributed to its uniqueness. However, fundamental differences even exist between distinct mammalian species. Here, we shall discuss the issue of our humanity from a neurobiological and historical perspective. PMID- 21647211 TI - The timing and strength of regional brain activation associated with word recognition in children with reading difficulties. AB - The study investigates the relative degree and timing of cortical activation across parietal, temporal, and frontal regions during performance of a continuous visual-word recognition task in children who experience reading difficulties (N = 44, RD) and typical readers (N = 40, NI). Minimum norm estimates of regional neurophysiological activity were obtained from magnetoencephalographic recordings. Children with RD showed bilaterally reduced neurophysiological activity in the superior and middle temporal gyri, and increased activity in rostral middle frontal and ventral occipitotemporal cortices, bilaterally. The temporal profile of activity in the RD group, featured near-simultaneous activity peaks in temporal, inferior parietal, and prefrontal regions, in contrast to a clear temporal progression of activity among these areas in the NI group. These results replicate and extend previous MEG and fMRI results demonstrating atypical, latency-dependent attributes of the brain circuit involved in word reading in children with reading difficulties. PMID- 21647213 TI - Feasibility of Repurposing the Polyanionic Microbicide, PPCM, for Prophylaxis against HIV Transmission during ART. AB - HIV-serodiscordant couples wishing to conceive often seek assisted reproduction, during which spermatozoa from infected men are washed to minimize the risk of HIV transmission to partner and fetus. We sought to improve this method by adding a microbicide, PPCM, as an HIV prophylactic. HIV-1 (BaL) inhibition by PPCM appears irreversible and independent of added Ca(2+). Without added Ca(2+), PPCM (<=10 mg/mL, <=90 min), a stimulus of Ca(2+)-dependent acrosomal loss, has no effect on sperm motility, forward progression, or acrosomal status. PPCM-treated (10 mg/mL) sperm retain their ability to acrosome react when Ca(2+) is added. Sperm DNA integrity/function is unaffected by PPCM (<=10 mg/mL). Adding PPCM (5 mg/mL, 30 min) to washing media reduces infectivity (viral antigen p24 and RNA) of ex-vivo HIV-infected semen by 3-4 Logs compared with washing alone. Sperm washing with appropriate extracellular Ca(2+) levels and PPCM is significantly more effective than washing alone at reducing HIV infectivity. PMID- 21647214 TI - Quantitative CT: Associations between Emphysema, Airway Wall Thickness and Body Composition in COPD. AB - The objective of the present study was to determine the association between CT phenotypes-emphysema by low attenuation area and bronchitis by airway wall thickness-and body composition parameters in a large cohort of subjects with and without COPD. In 452 COPD subjects and 459 subjects without COPD, CT scans were performed to determine emphysema (%LAA), airway wall thickness (AWT-Pi10), and lung mass. Muscle wasting based on FFMI was assessed by bioelectrical impedance. In both the men and women with COPD, FFMI was negatively associated with %LAA. FMI was positively associated with AWT-Pi10 in both subjects with and without COPD. Among the subjects with muscle wasting, the percentage emphysema was high, but the predictive value was moderate. In conclusion, the present study strengthens the hypothesis that the subgroup of COPD cases with muscle wasting have emphysema. Airway wall thickness is positively associated with fat mass index in both subjects with and without COPD. PMID- 21647215 TI - Analgesic efficacy of CR4056, a novel imidazoline-2 receptor ligand, in rat models of inflammatory and neuropathic pain. AB - Two decades of investigations have failed to unequivocally clarify the functions and the molecular nature of imidazoline-2 receptors (I2R). However, there is robust pharmacological evidence for the functional modulation of monoamino oxidase (MAO) and other important enzyme activities by I2 site ligands. Some compounds of this class proved to be active experimental tools in preventing both experimental pain and opioid tolerance and dependence. Unfortunately, even though these compounds bind with high potency to central I2 sites, they fail to represent a valid clinical opportunity due to their pharmacokinetic, selectivity or side-effects profile. This paper presents the preclinical profile of a novel I2 ligand (2-phenyl-6-(1H-imidazol-1yl) quinazoline; [CR4056]) that selectively inhibits the activity of human recombinant MAO-A in a concentration-dependent manner. A sub-chronic four day oral treatment of CR4056 increased norepinephrine (NE) tissue levels both in the rat cerebral cortex (63.1% +/-4.2%; P < 0.05) and lumbar spinal cord (51.3% +/- 6.7%; P < 0.05). In the complete Freund's adjuvant (CFA) rat model of inflammatory pain, CR4056 was found to be orally active (ED50 = 5.8 mg/kg, by mouth [p.o.]). In the acute capsaicin model, CR4056 completely blocked mechanical hyperalgesia in the injured hind paw (ED50 = 4.1 mg/kg, p.o.; ED100 = 17.9 mg/kg, p.o.). This effect was dose-dependently antagonized by the non-selective imidazoline I2/alpha2 antagonist idazoxan. In rat models of neuropathic pain, oral administration of CR4056 significantly attenuated mechanical hyperalgesia and allodynia. In summary, the present study suggests a novel pharmacological opportunity for inflammatory and/or neuropathic pain treatment based on selective interaction with central imidazoline-2 receptors. PMID- 21647216 TI - Evaluation of a local ICU sedation guideline on goal-directed administration of sedatives and analgesics. AB - PURPOSE: Sedatives and analgesics are commonly used in mechanically ventilated patients in the intensive care unit. Sedation guidelines have been shown to improve sedation management as well as various patient outcomes. The main objective was to evaluate adherence to a sedation guideline with both sedative prescribing and documentation of Richmond Agitation-Sedation Scale (RASS) scores. METHODS: In a retrospective chart review, data was collected on 111 medical intensive care unit patients mechanically ventilated via endotracheal tube for 12 hours or greater at Brigham and Women's Hospital. Fifty-seven patients were evaluated pre-guideline implementation and 54 patients were evaluated post guideline. RESULTS: Significant increases were seen in the post-guideline group in goal-directed sedation with a patient-specific RASS goal in the sedation order: 21.3 vs 85.4% (P < 0.001), and mean number of sedation assessments per 24 hours using the RASS: 4.7 vs 11.4 (P < 0.001). Similarly, this group experienced a higher percentage of RASS scores at their sedation goal: 31.4 vs 44.1% (P < 0.001). No difference was seen in other clinical endpoints. CONCLUSION: Implementation and routine application of a hospital pain and sedation guideline was associated with significantly improved sedation metrics, such as goal directed sedation, as well as frequency of sedation level assessment and documentation. An increase was observed in the time that post-guideline patients spent at or near their RASS goal. PMID- 21647217 TI - Computer-aided surface estimation of pain drawings - intra- and inter-rater reliability. AB - Pain drawings are often utilized in the documentation of pain conditions. The aim here was to investigate intra- and inter-rater reliability of area measurements performed on pain drawings consecutively, using the computer program Quantify One. Forty-eight patients with chronic nonmalignant pain had shaded in their experienced pain on the front and back views of a pain drawing. The templates were scanned and displayed on a 17-inch computer screen. Two independent examiners systematically encircled the shaded-in areas of the pain drawings with help of a computer mouse, twice each on two separate days, respectively. With this method it is possible to encircle each marked area and to obtain immediate details of its size. The total surface area (mm(2)) was calculated for each pain drawing measurement. Each examiner measured about 2400 areas, and as a whole, the number of areas measured varied only by 3%. The intra-rater reliability was high with intraclass correlation coefficients 0.992 in Examiner A and 0.998 in Examiner B. The intra-individual absolute differences were small within patients within one examiner as well as between the two examiners. The inter-rater reliability was also high. Still, significant differences in the absolute mean areas (13%) were seen between the two examiners in the second to fourth measurement sessions, indicating that one of the examiners measured systematically less. The measurement error was <=10%, indicating that use of the program would be advantageous both in clinical practice and in research, but if repeated, preferably with the same examiner. Since pain drawings with this method are digitized, high quality data without loss of information is possible to store in electronic medical records for later analysis, both regarding precise location and size of pain area. We conclude that the computer program Quantify One is a reliable method to calculate the areas of pain drawings. PMID- 21647218 TI - Primary somatosensory cortex in chronic low back pain - a H-MRS study. AB - The goal of this study was to investigate whether certain metabolites, specific to neurons, glial cells, and the neuronal-glial neurotransmission system, in the primary somatosensory cortex (SSC), are altered and correlated with clinical characteristics of pain in patients with chronic low back pain (LBP). Eleven LBP patients and eleven age-matched healthy controls were included. N-acetylaspartate (NAA), choline (Cho), myo-inositol (mI), and glutamine/glutamate (Glx) were measured with proton magnetic resonance spectroscopy ((1)H-MRS) in left and right SSC. Differences in metabolite concentrations relative to those of controls were evaluated as well as analyses of metabolite correlations within and between SSCs. Relationships between metabolite concentrations and pain characteristics were also evaluated. We found decreased NAA in the left SSC (P = 0.001) and decreased Cho (P = 0.04) along with lower correlations between all metabolites in right SSC (P = 0.007) in LBP compared to controls. In addition, we found higher and significant correlations between left and right mI (P < 0.001 in LBP vs P = 0.1 in controls) and between left mI and right Cho (P = 0.048 vs P = 0.6). Left and right NAA levels were negatively correlated with pain duration (P = 0.04 and P = 0.02 respectively) while right Glx was positively correlated with pain severity (P = 0.04). Our preliminary results demonstrated significant altered neuronal glial interactions in SSC, with left neural alterations related to pain duration and right neuronal-glial alterations to pain severity. Thus, the (1)H-MRS approach proposed here can be used to quantify relevant cerebral metabolite changes in chronic pain, and consequently increase our knowledge of the factors leading from these changes to clinical outcomes. PMID- 21647219 TI - Successful MRI-Guided Focused Ultrasound Uterine Fibroid Treatment Despite an Ostomy and Significant Abdominal Wall Scarring. AB - We present a case of successful magnetic resonance imaging-guided focused ultrasound surgery (MRgFUS) of a uterine fibroid in a patient with extensive anterior abdominal wall surgical scars from two longitudinal laparotomies, a total colectomy and ileostomy. This case demonstrates that MRgFUS can be safely used in patients with an ostomy and significant abdominal wall scarring, but careful pretreatment planning and positioning during treatment is needed. PMID- 21647220 TI - Knowledge, Attitudes, and Practice among Women and Doctors Concerning the Use of Folic Acid. AB - Background and Objective. Daily folic acid intake, prior to conception and in early pregnancy, significantly reduces neural tube defects (NTDs). We compared folic acid consumption among Jewish and Bedouin women and the recommendations of family physicians and gynecologists. Methods. We compared 64 Muslim Bedouin women and 65 Jewish women. We also compared 39 gynecologists and 60 family physicians. Results. Fifty-one Jewish women (78.5%) took folic acid during pregnancy, but only seven (10.8%) before conception. Sixty Bedouin women (93.75%) took folic acid during pregnancy, but only four (6.25%) before conception (P < .05). Five Jewish women (7.7%) and two Bedouin women (3.1%) took folic acid three months before conception. Thirty-three gynecologists (87%) recommend preconception folic acid compared with thirty-six family physicians (60%) (P < .05). Conclusions. The majority of women use folic acid during pregnancy, but only few do so to prevent NTDs. There is a significant difference between doctors' recommendations and actual practice. PMID- 21647221 TI - Gender difference of unconscious attentional bias in high trait anxiety individuals. AB - By combining binocular suppression technique and a probe detection paradigm, we investigated attentional bias to invisible stimuli and its gender difference in both high trait anxiety (HTA) and low trait anxiety (LTA) individuals. As an attentional cue, happy or fearful face pictures were presented to HTAs and LTAs for 800 ms either consciously or unconsciously (through binocular suppression). Participants were asked to judge the orientation of a gabor patch following the face pictures. Their performance was used to measure attentional effect induced by the cue. We found gender differences of attentional effect only in the unconscious condition with HTAs. Female HTAs exhibited difficulty in disengaging attention from the location where fearful faces were presented, while male HTAs showed attentional avoidance of it. Our results suggested that the failure to find attentional avoidance of threatening stimuli in many previous studies might be attributed to consciously presented stimuli and data analysis regardless of participants' gender. These findings also contributed to our understanding of gender difference in anxiety disorder. PMID- 21647222 TI - Prophylactic effect of a therapeutic vaccine against TB based on fragments of Mycobacterium tuberculosis. AB - The prophylactic capacity of the RUTI(r) vaccine, based on fragmented cells of Mycobacterium tuberculosis, has been evaluated in respect to aerosol challenge with virulent bacilli. Subcutaneous vaccination significantly reduced viable bacterial counts in both lungs and spleens of C57Bl mice, when challenged 4 weeks after vaccination. RUTI(r) protected the spleen less than BCG. Following a 9 month vaccination-challenge interval, protection was observed for the lungs, but not for the spleen. Survival of infected guinea pigs was prolonged by vaccination given 5 weeks before challenge. Inoculations of RUTI(r) shortly after infection significantly reduced the viable bacterial counts in the lungs, when compared with infected control mice. Thus, vaccination by RUTI(r) has potential for both the prophylaxis and immunotherapy of tuberculosis. PMID- 21647223 TI - The anti-inflammatory and antibacterial basis of human omental defense: selective expression of cytokines and antimicrobial peptides. AB - BACKGROUND: The wound healing properties of the human omentum are well known and have extensively been exploited clinically. However, the underlying mechanisms of these effects are not well understood. We hypothesize that the omentum tissue promotes wound healing via modulation of anti-inflammatory pathways, and because the omentum is rich in adipocytes, the adipocytes may modulate the anti inflammatory response. Factors released by human omentum may affect healing, inflammation and immune defense. METHODOLOGY: Six human omentum tissues (non obese, free from malignancy, and any other systemic disorder) were obtained during diagnostic laparoscopies having a negative outcome. Healthy oral mucosa (obtained from routine oral biopsies) was used as control. Cultured adipocytes derived from human omentum were exposed to lipopolysaccharide (LPS) (1-50 ng/mL) for 12-72 hours to identify the non-cytotoxic doses. Levels of expression (mRNA and protein) were carried out for genes associated with pro- and anti inflammatory cytokine responses and antibacterial/antimicrobial activity using qRT-PCR, western blotting, and cell-based ELISA assays. RESULTS: The study shows significant higher levels of expression (mRNA and protein) of several specific cytokines, and antibacterial peptides in the omentum tissues when compared to oral sub-mucosal tissues. In the validation studies, primary cultures of adipocytes, derived from human omentum were exposed to LPS (5 and 10 ng/mL) for 24 and 48 h. The altered expressions were more pronounced in cultured adipocytes cells when exposed to LPS as compared to the omentum tissue. CONCLUSIONS/SIGNIFICANCE: Perhaps, this is the first report that provides evidence of expressional changes in pro- and anti-inflammatory cytokines and antibacterial peptides in the normal human omentum tissue as well as adipocytes cultured from this tissue. The study provides new insights on the molecular and cellular mechanisms of healing and defense by the omentum, and suggests the potential applicability of cultured adipocytes derived from the omentum for future therapeutic applications. PMID- 21647224 TI - AC-NP: a novel chimeric peptide with natriuretic and vasorelaxing actions. AB - The aim of this study was to evaluate the cardiovascular and renal activities of a newly designed natriuretic peptide (NP). Here, we engineered a novel 28-amino acid chimeric peptide, termed AC-NP that combined the 17-amino acid ring of C type natriuretic peptide (CNP) with the 6-amino acid N-terminus and 5-amino acid C-terminus of atrial natriuretic peptide (ANP). Both in vitro and in vivo experiments were performed to determine the actions of AC-NP. In normal rats, AC NP proved to be more potentially diuretic, natriuretic and hypotensive compared with other NPs, such as ANP, CNP and vasonatrin peptide (VNP), which is another man-made NP. In relaxation of isolated abdominal aorta from rat, AC-NP was equally effective to ANP, CNP and VNP. Elevated levels of 3',5'-guanosine monophosphate (cGMP) in plasma and urine cGMP excretion indicated the participation of cGMP in the functions of AC-NP. Taken together, innovative designed AD-NP might be a new candidate therapeutic peptide against cardiorenal disorders. PMID- 21647225 TI - The mouse primary visual cortex is a site of production and sensitivity to estrogens. AB - The classic female estrogen, 17beta-estradiol (E2), has been repeatedly shown to affect the perceptual processing of visual cues. Although gonadal E2 has often been thought to influence these processes, the possibility that central visual processing may be modulated by brain-generated hormone has not been explored. Here we show that estrogen-associated circuits are highly prevalent in the mouse primary visual cortex (V1). Specifically, we cloned aromatase, a marker for estrogen-producing neurons, and the classic estrogen receptors (ERs) ERalpha and ERbeta, as markers for estrogen-responsive neurons, and conducted a detailed expression analysis via in-situ hybridization. We found that both monocular and binocular V1 are highly enriched in aromatase- and ER-positive neurons, indicating that V1 is a site of production and sensitivity to estrogens. Using double-fluorescence in-situ hybridization, we reveal the neurochemical identity of estrogen-producing and -sensitive cells in V1, and demonstrate that they constitute a heterogeneous neuronal population. We further show that visual experience engages a large population of aromatase-positive neurons and, to a lesser extent, ER-expressing neurons, suggesting that E2 levels may be locally regulated by visual input in V1. Interestingly, acute episodes of visual experience do not affect the density or distribution of estrogen-associated circuits. Finally, we show that adult mice dark-reared from birth also exhibit normal distribution of aromatase and ERs throughout V1, suggesting that the implementation and maintenance of estrogen-associated circuits is independent of visual experience. Our findings demonstrate that the adult V1 is a site of production and sensitivity to estrogens, and suggest that locally-produced E2 may shape visual cortical processing. PMID- 21647226 TI - The endoplasmic reticulum chaperone protein GRP94 is required for maintaining hematopoietic stem cell interactions with the adult bone marrow niche. AB - Hematopoietic stem cell (HSC) homeostasis in the adult bone marrow (BM) is regulated by both intrinsic gene expression products and interactions with extrinsic factors in the HSC niche. GRP94, an endoplasmic reticulum chaperone, has been reported to be essential for the expression of specific integrins and to selectively regulate early T and B lymphopoiesis. In GRP94 deficient BM chimeras, multipotent hematopoietic progenitors persisted and even increased, however, the mechanism is not well understood. Here we employed a conditional knockout (KO) strategy to acutely eliminate GRP94 in the hematopoietic system. We observed an increase in HSCs and granulocyte-monocyte progenitors in the Grp94 KO BM, correlating with an increased number of colony forming units. Cell cycle analysis revealed that a loss of quiescence and an increase in proliferation led to an increase in Grp94 KO HSCs. This expansion of the HSC pool can be attributed to the impaired interaction of HSCs with the niche, evidenced by enhanced HSC mobilization and severely compromised homing and lodging ability of primitive hematopoietic cells. Transplanting wild-type (WT) hematopoietic cells into a GRP94 null microenvironment yielded a normal hematology profile and comparable numbers of HSCs as compared to WT control, suggesting that GRP94 in HSCs, but not niche cells, is required for maintaining HSC homeostasis. Investigating this, we further determined that there was a near complete loss of integrin alpha4 expression on the cell surface of Grp94 KO HSCs, which showed impaired binding with fibronectin, an extracellular matrix molecule known to play a role in mediating HSC-niche interactions. Furthermore, the Grp94 KO mice displayed altered myeloid and lymphoid differentiation. Collectively, our studies establish GRP94 as a novel cell intrinsic factor required to maintain the interaction of HSCs with their niche, and thus regulate their physiology. PMID- 21647227 TI - Towards the human colorectal cancer microbiome. AB - Multiple factors drive the progression from healthy mucosa towards sporadic colorectal carcinomas and accumulating evidence associates intestinal bacteria with disease initiation and progression. Therefore, the aim of this study was to provide a first high-resolution map of colonic dysbiosis that is associated with human colorectal cancer (CRC). To this purpose, the microbiomes colonizing colon tumor tissue and adjacent non-malignant mucosa were compared by deep rRNA sequencing. The results revealed striking differences in microbial colonization patterns between these two sites. Although inter-individual colonization in CRC patients was variable, tumors consistently formed a niche for Coriobacteria and other proposed probiotic bacterial species, while potentially pathogenic Enterobacteria were underrepresented in tumor tissue. As the intestinal microbiota is generally stable during adult life, these findings suggest that CRC associated physiological and metabolic changes recruit tumor-foraging commensal like bacteria. These microbes thus have an apparent competitive advantage in the tumor microenvironment and thereby seem to replace pathogenic bacteria that may be implicated in CRC etiology. This first glimpse of the CRC microbiome provides an important step towards full understanding of the dynamic interplay between intestinal microbial ecology and sporadic CRC, which may provide important leads towards novel microbiome-related diagnostic tools and therapeutic interventions. PMID- 21647228 TI - Unusual contents of the femoral hernia. AB - Different contents in the femoral hernia have been reported in the literature, but herniation of the fallopian tube in a femoral hernia is very rare due to its normal anatomical position. Case Presentation. A female patient was admitted to the surgical ward for a lump in the right groin. Clinical examination confirmed a right femoral hernia. The patient underwent surgery to repair the hernia. Intraoperatively, the right uterine tube was found in the hernia. The tube was reduced back into the pelvic cavity and the hernia was repaired. After making good recovery, the patient was referred to the gynaecologist for further assessment. Conclusion. This case is educational as it highlights the importance of managing women with femoral masses with care. PMID- 21647229 TI - Laparoscopic approach of a unicornuate uterus with noncommunicating rudimentary horns. AB - Background. Mullerian duct malformations delineate a miscellaneous group of congenital anomalies that result from arrested development, abnormal formation, or incomplete fusion of the mesonephric ducts. Case. This paper describes the diagnosis and management of a noncommunicating rudimentary horn complicated by severe pelvic pain and associated endometriosis. Conclusion. This condition was diagnosed by laparoscopy and hysteroscopy examination. Operative videolaparoscopy proved to be a successful approach for the treatment of this congenital Mullerian anomaly. PMID- 21647230 TI - Effect of pregnancy and childbirth on sexuality of women in ibadan, Nigeria. AB - A study of 375 antenatal attendees to assess women's views and experience in sexual matters during pregnancy and following childbirth. Explanatory variables included the perception women had of sex during pregnancy and after childbirth. Outcome variables were frequency and satisfaction of sexual activity. The commonest reasons for having coitus in pregnancy were marital harmony and facilitation of delivery. Libido rose throughout pregnancy but orgasms were less often experienced. The man-on-top position became less practised. Vaginal intercourse remained the commonest type. Masturbation and anal intercourse increased, while oral sex declined throughout. Marriage (OR 9.0, 95% CI 1.0-79.5) and current cohabitation (OR 13.6, 95% CI 1.6-113.4) were predictors of sex in pregnancy. Dyspareunia and partners' extramarital affairs were deterrent. Vaginal delivery and episiotomy were not significant predictors of postnatal sex. The respondents and their partners seem able to adapt to pregnancy changes and enhance their marital bonds. Anticipatory guidance and informed counselling may encourage this. PMID- 21647231 TI - Asymptomatic bacteriuria and antibacterial susceptibility patterns in an obstetric population. AB - Introduction. Asymptomatic bacteriuria (ASB), occurring in 2-11% of pregnancies, is a major predisposition to the development of pyelonephritis, which is associated with obstetrical complications, such as preterm labor and low birth weight infants. The aim of this study was to determine the prevalence of ASB, the antibacterial susceptibilities of the isolated microorganisms and the associated risk factors in an outpatient clinical setting in Zekai Tahir Burak Women's Health Education and Research Hospital in Ankara, Turkey. Material and Methods. Between December 2009 and May 2010, pregnant women admitted to the antenatal outpatient clinic were included in this study. The results of a complete urine analysis, midstream urine culture and antibacterial susceptibility were evaluated. Results. Of the 2011 pregnant women included, 171 had ASB (8.5%). E. coli was the most frequently isolated microorganism (76.6%), followed by Klebsiella pneumonia (14.6%). Both microorganisms were highly sensitive to fosfomycin, sensivity being 99.2% for E. coli and 88% for Klebsiella pneumonia. Conclusions. In this certain geographical region, we found E. coli as the most common causative agent of ASB in the obstetric population and it is very sensitive to fosfomycin. We recommend fosfomycin for ASB in pregnant women due to its high sensitivity, ease of administration and safety for use in pregnancy. PMID- 21647232 TI - Regulatory Effect of Peptidoglycan on the Expression of Toll-Like Receptor 2 mRNA and Proteins in Trophoblast Cell Line TEV-1 Cells. AB - Objective. To investigate the regulatory effect of peptidoglycan on the expression of human Toll-like receptors 2 (TLR2) mRNA and proteins in the human extravillous trophoblast cell line (TEV-1). Methods. TEV-1 cells were incubated with different doses of peptidoglycan. The expression of TLR2 mRNA and protein was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunocytochemistry SP staining. Results. TLR2 was expressed in TEV-1 cells and localized to both the cytoplasm and plasma membrane. Compared with the untreated control, TEV-1 cells incubated with 30 MUg/ml peptidoglycan significantly upregulated the expression of TLR2 mRNA and protein after 12 hours of treatment (P < .01). However, the expression of TLR2 mRNA and protein was decreased but had no significant difference compared with the control (P > .05) after 24 hours of treatment. On the other hand, 10 MUg/ml peptidoglycan did not seem to have regulatory effect on mRNA and protein expression of TLR2 (P > .05). Conclusion. Peptidoglycan has a role in regulating the expression of TLR2 mRNA and protein in TEV-1 cells. It suggests that the trophoblast cells may play important role in the immune response at the fetal-maternal interface and affect the result of pregnancy by expressing TLR2. PMID- 21647233 TI - Disseminated Intravascular Coagulopathy following Induced Second Trimester Curettage Abortion. AB - Background. This is a case of 18-year-old adolescent girl admitted with profuse vaginal bleeding following induced second trimester curettage abortion at 13 weeks of gestation. Case. Her transvaginal sonogram detected retained products of conceptus, and her blood reports revealed a full blown picture of DIC. Dilatation and evacuation was done after initial resuscitation with packed RBCs and platelet concentrates. She had an uneventful recovery period. Conclusion. DIC is an extremely life-threatening condition which can occur as a very infrequent complication of second trimester abortion. PMID- 21647234 TI - Clinicopathological characteristics and her-2/neu status in chinese patients with uterine papillary serous carcinoma. AB - Objective. To analyze clinico-pathological features of Chinese patients with UPSC, and investigate roles of Her-2/neu protein expression and gene amplification in UPSC prognosis. Methods. Thirty-six patients with UPSC treated in Cancer Hospital of Fudan University from 1996 to 2006 were analysed retrospectively. Chromogenic in situ hybridization (CISH) and immunohistochemistry (IHC) were performed to evaluate Her-2/neu gene amplification and protein expression respectively. Results. The median age was 63 years, and 61% (22/36) were late stages (stage III/IV). The 1-year, 3-year, and 5 year overall survival (OS) was 73.1%, 51.9% and 43.9%, respectively. Advanced stages (P = .0006) and deep myometrial invasion (P = .0138) were significantly associtated with a shorter OS. In 36 cases, 27.8% (10/36) showed 2+ staining and 8.3% (3/36) showed 3+ by IHC. Amplification of the Her-2/neu gene was observed in 11.1% (4/36) cases. The 5-year overall survival rate in Her-2/neu IHC 2 + ~3+ and 0 ~ 1+ cases was 12.9% and 68.6% respectively. Her-2/neu protein expression 2 + ~3+ was significantly associated with advanced surgical stage and worse overall survival (P = .03 and P = .0023, resp.). Conclusion. Chinese patients with UPSC showed characteristics of deep myometrial invasion, advanced stages and poor overall survival. Her-2/neu protein overexpression is associated with advanced stage and poor survival outcome. PMID- 21647236 TI - Highly diverse, poorly studied and uniquely threatened by climate change: an assessment of marine biodiversity on South Georgia's continental shelf. AB - We attempt to quantify how significant the polar archipelago of South Georgia is as a source of regional and global marine biodiversity. We evaluate numbers of rare, endemic and range-edge species and how the faunal structure of South Georgia may respond to some of the fastest warming waters on the planet. Biodiversity data was collated from a comprehensive review of reports, papers and databases, collectively representing over 125 years of polar exploration. Classification of each specimen was recorded to species level and fully geo referenced by depth, latitude and longitude. This information was integrated with physical data layers (e.g. temperature, salinity and flow) providing a visualisation of South Georgia's biogeography across spatial, temporal and taxonomic scales, placing it in the wider context of the Southern Hemisphere. This study marks the first attempt to map the biogeography of an archipelago south of the Polar Front. Through it we identify the South Georgian shelf as the most speciose region of the Southern Ocean recorded to date. Marine biodiversity was recorded as rich across taxonomic levels with 17,732 records yielding 1,445 species from 436 families, 51 classes and 22 phyla. Most species recorded were rare, with 35% recorded only once and 86% recorded <10 times. Its marine fauna is marked by the cumulative dominance of endemic and range-edge species, potentially at their thermal tolerance limits. Consequently, our data suggests the ecological implications of environmental change to the South Georgian marine ecosystem could be severe. If sea temperatures continue to rise, we suggest that changes will include depth profile shifts of some fauna towards cooler Antarctic Winter Water (90-150 m), the loss of some range-edge species from regional waters, and the wholesale extinction at a global scale of some of South Georgia's endemic species. PMID- 21647237 TI - Erratum: "Temperature dependence of 1/f noise mechanisms in silicon nanowire biochemical field effect transistors" [Appl. Phys. Lett. 97, 243501 (2010)]. AB - [This corrects the article on p. 243501 in vol. 97, PMID: 21221250.][This corrects the article on p. 243501 in vol. 97, PMID: 21221250.]. PMID- 21647238 TI - Tripeptidyl Peptidase II Is Required for c-MYC-Induced Centriole Overduplication and a Novel Therapeutic Target in c-MYC-Associated Neoplasms. AB - Centrosome aberrations are frequently detected in c-MYC-associated human malignancies. Here, we show that c-MYC-induced centrosome and centriole overduplication critically depend on the protease tripeptidyl peptidase II (TPPII). We found that TPPII localizes to centrosomes and that overexpression of TPPII, similar to c-MYC, can disrupt centriole duplication control and cause centriole multiplication, a process during which maternal centrioles nucleate the formation of more than a single daughter centriole. We report that inactivation of TPPII using chemical inhibitors or siRNA-mediated protein knockdown effectively reduced c-MYC-induced centriole overduplication. Remarkably, the potent and selective TPPII inhibitor butabindide not only potently suppressed centriole aberrations but also caused significant cell death and growth suppression in aggressive human Burkitt lymphoma cells with c-MYC overexpression. Taken together, these results highlight the role of TPPII in c-MYC-induced centriole overduplication and encourage further studies to explore TPPII as a novel antineoplastic drug target. PMID- 21647239 TI - Transnational Home Engagement among Latino and Asian Americans: Resources and Motivation. AB - Is immigrant groups' assimilation to host society at odds with their engagement with the country of ancestral origin? This study divides the concept of assimilation into socioeconomic resources and attachment to host society, and argues that assimilation and transnational perspectives are coexisting paradigms. Analyses using the nationally representative samples of Latino and Asian Americans indicate that 1) higher-order generations reduce the odds of home country engagement, i.e. frequent return visits, 2) attachment to American society does not discourage return visits, 3) socioeconomic resources increase frequent visits, and 4) the country of origin is a significant predictor of home country visits. PMID- 21647240 TI - Attachment Anxiety, Verbal Immediacy, and Blood Pressure: Results from a Laboratory-Analogue Study Following Marital Separation. AB - Marital separation and divorce increase risk for all-cause morbidity and mortality. Using a laboratory analogue paradigm, the present study examined attachment anxiety, language use, and blood pressure (BP) reactivity among 119 (n = 43 men, 76 women) recently separated adults who were asked to mentally reflect on their relationship history and separation experience. We created a language use composite of verbal immediacy from participants' stream-of-consciousness recordings about their separation experience as a behavioral index of attachment related hyperactivation. Verbal immediacy moderated the association between attachment anxiety and BP at the beginning of a divorce-specific activation task. Participants reporting high attachment anxiety who discussed their separation in a first-person, present-oriented and highly engaged manner evidenced the highest levels of BP at the start of the divorce-specific task. Results provide a deeper understanding of the association between marital dissolution and health and suggest that verbal immediacy may be a useful behavioral index of hyperactivating coping strategies. PMID- 21647241 TI - Understanding links between punitive parenting and adolescent adjustment: The relevance of context and reciprocal associations. AB - There is considerable debate regarding the extent to which punitive parenting adversely impacts youth well being. Using an ecological-transactional model of human development, we examined reciprocity and contextual variability in associations between maternal punitive discipline and adolescent adjustment among 1,147 low-income, urban youth followed through adolescence. Longitudinal SEM results indicated that delinquency and depressive symptoms during pre- and early adolescence (Time 1) were associated with increased punitive discipline about a year later (Time 2). When mothers reported less Time 2 neighborhood disorder, punitive discipline at Time 2 was associated with increased delinquency (for boys) and depressive symptoms (for girls) during mid- to late adolescence (Time 3). The costs of punitive discipline for adolescent adjustment are best understood considering the dynamic, transactional, and contextual nature of development. PMID- 21647242 TI - An Entropy Approach to Disclosure Risk Assessment: Lessons from Real Applications and Simulated Domains. AB - We live in an increasingly mobile world, which leads to the duplication of information across domains. Though organizations attempt to obscure the identities of their constituents when sharing information for worthwhile purposes, such as basic research, the uncoordinated nature of such environment can lead to privacy vulnerabilities. For instance, disparate healthcare providers can collect information on the same patient. Federal policy requires that such providers share "de-identified" sensitive data, such as biomedical (e.g., clinical and genomic) records. But at the same time, such providers can share identified information, devoid of sensitive biomedical data, for administrative functions. On a provider-by-provider basis, the biomedical and identified records appear unrelated, however, links can be established when multiple providers' databases are studied jointly. The problem, known as trail disclosure, is a generalized phenomenon and occurs because an individual's location access pattern can be matched across the shared databases. Due to technical and legal constraints, it is often difficult to coordinate between providers and thus it is critical to assess the disclosure risk in distributed environments, so that we can develop techniques to mitigate such risks. Research on privacy protection has so far focused on developing technologies to suppress or encrypt identifiers associated with sensitive information. There is growing body of work on the formal assessment of the disclosure risk of database entries in publicly shared databases, but a less attention has been paid to the distributed setting. In this research, we review the trail disclosure problem in several domains with known vulnerabilities and show that disclosure risk is influenced by the distribution of how people visit service providers. Based on empirical evidence, we propose an entropy metric for assessing such risk in shared databases prior to their release. This metric assesses risk by leveraging the statistical characteristics of a visit distribution, as opposed to person-level data. It is computationally efficient and superior to existing risk assessment methods, which rely on ad hoc assessment that are often computationally expensive and unreliable. We evaluate our approach on a range of location access patterns in simulated environments. Our results demonstrate the approach is effective at estimating trail disclosure risks and the amount of self-information contained in a distributed system is one of the main driving factors. PMID- 21647243 TI - Water Permeation Through DMPC Lipid Bilayers using Polarizable Charge Equilibration Force Fields. AB - We investigate permeation energetics of water entering a model dimyristoylphosphatidylcholine (DMPC) bilayer via molecular dynamics simulations using polarizable Charge Equilibration (CHEQ) models. Potentials of mean force show 4.5-5.5 kcal/mol barriers for water permeation into bilayers. Barriers are highest when water coordination within the bilayer is prevented, and also when using force fields that accurately reproduce experimental alkane hydration free energies. The magnitude of the average water dipole moment decreases from 2.6 Debye (in bulk) to 1.88 Debye (in membrane interior). This variation correlates with the change in a water molecule's coordination number. PMID- 21647244 TI - Design issues for population growth models. AB - We briefly review and discuss design issues for population growth and decline models. We then use a flexible growth and decline model as an illustrative example and apply optimal design theory to find optimal sampling times for estimating model parameters, specific parameters and interesting functions of the model parameters for the model with two real applications. Robustness properties of the optimal designs are investigated when nominal values or the model is mis specified, and also under a different optimality criterion. To facilitate use of optimal design ideas in practice, we also introduce a website for generating a variety of optimal designs for popular models from different disciplines. PMID- 21647245 TI - ASSESSMENT OF DISTRESS IN YOUNG CHILDREN: A COMPARISON OF AUTISTIC DISORDER, DEVELOPMENTAL DELAY, AND TYPICAL DEVELOPMENT. AB - Distress emotions in very young children are manifest in vocal, facial, and bodily cues. Moreover, children with different developmental conditions (i.e. Autistic Disorder- AD, Developmental Delay- DD, Typically Developing- TD) appear to manifest their distress emotions via different channels. To decompose channel of emotional distress display by group, we conducted a study in which video clips of crying of 18 children 18 months of age belonging to three groups (AD, DD, TD) were modified to isolate vocal, facial, or bodily cues, and 42 female adults were asked to judge the distress and typicality (expected normality) of the different stimuli. We find variation in adult judgements of distress and typicality by child group (AD, DD, TD) and by isolated cues (vocal, facial, or body). Although there is some overlap between responses to episodes of crying of children with AD and those with DD, the different cues of crying of children with AD tend to be considered more atypical and distressed than those of the other two groups (DD and TD). Early assessment of different cues of the expression of distress, and more generally of emotional expressivity in a child, may provide useful information for pediatricians and practitioners who are in contact with young children and must make clinical screening decisions. The findings also alert parents of children with AD to important aspects of their cries. PMID- 21647246 TI - An adaptive Dynamic Relaxation method for solving nonlinear finite element problems. Application to brain shift estimation. AB - Dynamic Relaxation is an explicit method that can be used for computing the steady state solution for a discretised continuum mechanics problem. The convergence speed of the method depends on the accurate estimation of the parameters involved, which is especially difficult for nonlinear problems. In this paper we propose a completely adaptive Dynamic Relaxation method in which the parameters are updated during the iteration process, converging to their optimal values. We use the proposed method for computing intra-operative organ deformations using non-linear finite element models involving large deformations, nonlinear materials and contacts. The simulation results prove the accuracy and computational efficiency of the method. The proposed method is also very well suited for GPU implementation. PMID- 21647247 TI - Disparities in the moral intuitions of criminal offenders: The role of psychopathy. AB - The present study examined whether and in what ways psychopathy is associated with abnormal moral intuitions among criminal offenders. Using Haidt et al.'s Moral Foundations Questionnaire, 222 adult male offenders assessed for clinical psychopathy reported their degree of support for five moral domains: Harm Prevention, Fairness, Respect for Authority, Ingroup Loyalty, and Purity/Sanctity. As predicted, psychopathy total score explained a substantial proportion of the variance in reduced support for Harm Prevention and Fairness, but not the other domains. These results confirm that psychopathy entails a discrete set of moral abnormalities and suggest that these abnormalities could potentially help to explain the characteristic antisocial behavior of individuals with psychopathy. PMID- 21647248 TI - Ultracompact beam splitters based on plasmonic nanoslits. AB - An ultracompact plasmonic beam splitter is theoretically and numerically investigated. The splitter consists of a V-shaped nanoslit in metal films. Two groups of nanoscale metallic grooves inside the slit (A) and at the small slit opening (B) are investigated. We show that there are two energy channels guiding light out by the splitter: the optical and the plasmonic channels. Groove A is used to couple incident light into the plasmonic channel. Groove B functions as a plasmonic scatter. We demonstrate that the energy transfer through plasmonic path is dominant in the beam splitter. We find that more than four times the energy is transferred by the plasmonic channel using structures A and B. We show that the plasmonic waves scattered by B can be converted into light waves. These light waves redistribute the transmitted energy through interference with the field transmitted from the nanoslit. Therefore, different beam splitting effects are achieved by simply changing the interference conditions between the scattered waves and the transmitted waves. The impact of the width and height of groove B are also investigated. It is found that the plasmonic scattering of B is changed into light scattering with increase of the width and the height of B. These devices have potential applications in optical sampling, signal processing, and integrated optical circuits. PMID- 21647249 TI - SRC-3 has a role in cancer other than as a nuclear receptor coactivator. AB - Steroid receptor coactivator-3 (SRC-3), also known as AIB1, is a member of the p160 steroid receptor coactivator family. Since SRC-3 was found to be amplified in breast cancer in 1997, the role of SRC-3 in cancer has been broadly investigated. SRC-3 initially was identified as a transcriptional coactivator for nuclear receptors such as the estrogen receptor (ER), involved in the proliferation of hormone-dependent cancers. However, increasing clinical evidence shows that dysregulation of SRC-3 expression in several human hormone-independent cancers is correlated with pathological factors and clinical prognosis. Recently, both in vivo and in vitro studies demonstrate that SRC-3 may influence a number of cancer cellular processes in several ways independent of nuclear receptor signaling. In addition, an SRC-3 transgenic mice model shows that SRC-3 induces tumors in several mouse tissues. These results indicate that the role of SRC-3 in cancer is not just as a nuclear receptor coactivator. The focus of this review is to examine possible SRC-3 roles in cancer, other than as a nuclear receptor coactivator. PMID- 21647250 TI - Berberine regulated Gck, G6pc, Pck1 and Srebp-1c expression and activated AMP activated protein kinase in primary rat hepatocytes. AB - The effects of hormonal and dietary stimuli on hepatic glucose and lipid homeostasis include regulation of gene expression. Berberine, an effective compound in certain Chinese medicinal herbs, has been reported to lower plasma glucose and lipid levels in diabetic and hypercholesterolemic patients. We hypothesized that it may affect the expression of hepatic genes involved in glucose and lipid metabolism. The effects of berberine hydrochloride on viability, gene expression, and activation of AMP activated protein kinase (AMPK) in primary hepatocytes from Sprague-Dawley (SD), Zucker lean (ZL) or fatty (ZF) rats were examined with MTT assay, real-time PCR, and western blotting, respectively. Berberine hydochloride at 50 uM or higher caused cytotoxic effects on hepatocytes. In SD and ZL hepatocytes, it induced Gck and suppressed G6pc expression at 10 and 25 uM, but not as potent as 1 nM insulin. Its effects on Pck1, and insulin-regulated Gck and G6pc expression depended on the hepatocyte sources and the dosage used. In ZF hepatocytes, it increased Gck, and suppressed Pck1 and G6pc expression without insulin. Its effects on Gck and G6pc, but not Pck1 expression, were additive with insulin. Berberine hydrochloride at 25 uM attenuated insulin-suppressed Pck1 (ZL/ZF cells), and insulin-induced Srebp-1c expression (SD/ZL/ZF cells), suggesting modulation of insulin action. Berberine hydrochloride did not alter these genes' mRNA stability. Its treatment caused a dose-dependent increase of phosphorylation of AMPKalpha, and its substrate, acetyl-CoA carboxylase, in primary hepatocytes. We conclude that berberine hydrochloride regulated the transcription of hepatic genes involved in glucose and fatty acid metabolism. PMID- 21647252 TI - Gold catalysis for organic synthesis. PMID- 21647251 TI - miR-21 promotes keratinocyte migration and re-epithelialization during wound healing. AB - MicroRNAs involved in keratinocyte migration and wound healing are largely unknown. Here, we revealed the indispensable role of miR-21 in keratinocyte migration and in re-epithelialization during wound healing in mice. In HaCaT cell, miR-21 could be upregulated by TGF-beta1. Similar to the effect of TGF beta1, miR-21 overexpression promoted keratinocyte migration. Conversely, miR-21 knockdown attenuated TGF-beta1-induced keratinocyte migration, suggesting that miR-21 was essential for TGF-beta-driven keratinocyte migration. Furthermore, we found that miR-21 was upregulated during wound healing, coincident with the temporal expression pattern of TGF-beta1. Consistently, knockdown of endogenous miR-21 using a specific antagomir dramatically delayed re-epithelialization possibly due to the reduced keratinocyte migration. TIMP3 and TIAM1, direct targets of miR-21, were verified to be regulated by miR-21 in vitro and in vivo, indicating that these two molecules might contribute to miR-21-induced keratinocyte migration. Taken together, our results demonstrate that miR-21 promotes keratinocyte migration and boosts re-epithelialization during skin wound healing. PMID- 21647253 TI - Novel carbazole-pyridine copolymers by an economical method: synthesis, spectroscopic and thermochemical studies. AB - The synthesis, as well as spectroscopic and thermochemical studies of a novel class of carbazole-4-phenylpyridine co-polymers are described. The synthesis was carried out by a simple and cheaper method compared to the lengthy methods usually adopted for the preparation of carbazole-pyridine copolymers which involve costly catalysts. Thus, two series of polymers were synthesized by a modified Chichibabin reaction, i.e., by the condensation of diacetylated N alkylcarbazoles with 3-substituted benzaldehydes in the presence of ammonium acetate in refluxing acetic acid. All the polymers were characterized by FTIR, (1)H NMR, (13)C NMR, UV-vis spectroscopy, fluorimetry, TGA and DSC. The weight average molecular masses (M(w)) of the polymers were estimated by the laser light scattering (LLS) technique. PMID- 21647254 TI - Sequential Au(I)-catalyzed reaction of water with o-acetylenyl-substituted phenyldiazoacetates. AB - The gold(I)-catalyzed reaction of water with o-acetylenyl-substituted phenyldiazoacetates provides 1H-isochromene derivatives in good yields. The reaction follows a catalytic sequence of gold carbene formation/water O-H insertion/alcohol-alkyne cyclization. The gold(I) complex is the only catalyst in each of these steps. PMID- 21647255 TI - Kinetic evaluation of the solvolysis of isobutyl chloro- and chlorothioformate esters. AB - The specific rates of solvolysis of isobutyl chloroformate (1) are reported at 40.0 degrees C and those for isobutyl chlorothioformate (2) are reported at 25.0 degrees C, in a variety of pure and binary aqueous organic mixtures with wide ranging nucleophilicity and ionizing power. For 1, we also report the first-order rate constants determined at different temperatures in pure ethanol (EtOH), methanol (MeOH), 80% EtOH, and in both 97% and 70% 2,2,2-trifluoroethanol (TFE). The enthalpy (DeltaH(?)) and entropy (DeltaS(?)) of activation values obtained from Arrhenius plots for 1 in these five solvents are reported. The specific rates of solvolysis were analyzed using the extended Grunwald-Winstein equation. Results obtained from correlation analysis using this linear free energy relationship (LFER) reinforce our previous suggestion that side-by-side addition elimination and ionization mechanisms operate, and the relative importance is dependent on the type of chloro- or chlorothioformate substrate and the solvent. PMID- 21647256 TI - Asymmetric synthesis of tertiary thiols and thioethers. AB - Enantiomerically pure tertiary thiols provide a major synthetic challenge, and despite the importance of chiral sulfur-containing compounds in biological and medicinal chemistry, surprisingly few effective methods are suitable for the asymmetric synthesis of tertiary thiols. This review details the most practical of the methods available. PMID- 21647257 TI - Predicting the UV-vis spectra of oxazine dyes. AB - In the current work we have investigated the ability of time-dependent density functional theory (TD-DFT) to predict the absorption spectra of a series of oxazine dyes and the effect of solvent on the accuracy of these predictions. Based on the results of this study, it is clear that for the series of oxazine dyes an accurate prediction of the excitation energy requires the inclusion of solvent. Implicit solvent included via a polarizable continuum approach was found to be sufficient in reproducing the excitation energies accurately in the majority of cases. Moreover, we found that the SMD solvent model, which is dependent on the full electron density of the solute without partitioning into partial charges, gave more reliable results for our systems relative to the conductor-like polarizable continuum model (CPCM), as implemented in Gaussian 09. In all cases the inclusion of solvent reduces the error in the predicted excitation energy to <0.3 eV and in the majority of cases to <0.1 eV. PMID- 21647258 TI - Intraannular photoreactions in pseudo-geminally substituted [2.2]paracyclophanes. AB - The photoisomerization of the pseudo-geminal tetraene 11 furnishes the cyclooctadiene derivatives 13 and 15 with a completely new type of molecular bridge for a [2.2]paracyclophane which promise many interesting novel applications; the same is true for the photoisomerization of 22 to 23 and 24. The structures of these new hydrocarbons were established by X-ray crystallography and spectroscopic analysis; among the noteworthy structural features of 13 and 15 are unusually long carbon-carbon single bonds (>1.64 A). PMID- 21647259 TI - Gold-catalyzed regioselective oxidation of terminal allenes: formation of alpha methanesulfonyloxy methyl ketones. AB - Synthetically useful alpha-methanesulfonyloxy methyl ketones are readily prepared in one-step from terminal allenes in fair to good yields. The chemistry relies on a gold-catalyzed intermolecular oxidation of the 1,2-diene unit using 3,5 dichloropyridine N-oxide as the oxidant. The reaction tolerates a range of functional groups and shows excellent regioselectivity. PMID- 21647260 TI - Synthesis of 5-(2-methoxy-1-naphthyl)- and 5-[2-(methoxymethyl)-1-naphthyl]-11H benzo[b]fluorene as 2,2'-disubstituted 1,1'-binaphthyls via benzannulated enyne allenes. AB - 5-(2-Methoxy-1-naphthyl)- and 5-[2-(methoxymethyl)-1-naphthyl]-11H benzo[b]fluorene were synthesized by treatment of the corresponding benzannulated enediynes with potassium tert-butoxide in refluxing toluene to give benzannulated enyne-allenes for the subsequent Schmittel cascade cyclization reactions. The structures of these two 5-(1-naphthyl)-11H-benzo[b]fluorenes could be regarded as 2,2'-disubstituted 1,1'-binaphthyls with the newly constructed benzofluorenyl group serving as a naphthyl moiety. PMID- 21647261 TI - Complete transfer of chirality in an intramolecular, thermal [2 + 2] cycloaddition of allene-ynes to form non-racemic spirooxindoles. AB - A thermal [2 + 2] cycloaddition reaction of allene-ynes has been used to transform chiral non-racemic allenyl oxindoles into chiral non-racemic spirooxindoles containing an alkylidene cyclobutene moiety. The enantiomeric excesses were determined by chiral lanthanide shift NMR analysis and the transfer of chiral information from the allene to the spirooxindole was found to be greater than 95%. PMID- 21647262 TI - An overview of the key routes to the best selling 5-membered ring heterocyclic pharmaceuticals. AB - This review presents a comprehensive overview on selected synthetic routes towards commercial drug compounds as published in both journal and patent literature. Owing to the vast number of potential structures, we have concentrated only on those drugs containing five-membered heterocycles and focused principally on the assembly of the heterocyclic core. In order to target the most representative chemical entities the examples discussed have been selected from the top 200 best selling drugs of recent years. PMID- 21647263 TI - The arene-alkene photocycloaddition. AB - In the presence of an alkene, three different modes of photocycloaddition with benzene derivatives can occur; the [2 + 2] or ortho, the [3 + 2] or meta, and the [4 + 2] or para photocycloaddition. This short review aims to demonstrate the synthetic power of these photocycloadditions. PMID- 21647264 TI - One-pot gold-catalyzed synthesis of 3-silylethynyl indoles from unprotected o alkynylanilines. AB - The Au(III)-catalyzed cyclization of 2-alkynylanilines was combined in a one-pot procedure with the Au(I)-catalyzed C3-selective direct alkynylation of indoles using the benziodoxolone reagent TIPS-EBX to give a mild, easy and straightforward entry to 2-substituted-3-alkynylindoles. The reaction can be applied to unprotected anilines, was tolerant to functional groups and easy to carry out (RT, and requires neither an inert atmosphere nor special solvents). PMID- 21647265 TI - Construction of cyclic enones via gold-catalyzed oxygen transfer reactions. AB - During the last decade, gold-catalyzed reactions have become a tour de force in organic synthesis. Recently, the gold-, Bronsted acid- or Lewis acid-catalyzed oxygen transfer from carbonyl to carbon-carbon triple bond, the so-called alkyne carbonyl metathesis, has attracted much attention because this atom economical transformation generates alpha,beta-unsaturated carbonyl derivatives which are of great interest in synthetic organic chemistry. This mini-review focuses on the most recent achievements on gold-catalyzed oxygen transfer reactions of tethered alkynones, diynes or alkynyl epoxides to cyclic enones. The corresponding mechanisms for the transformations are also discussed. PMID- 21647266 TI - Synthesis of cross-conjugated trienes by rhodium-catalyzed dimerization of monosubstituted allenes. AB - A rhodium(I)/dppe catalyst promoted dimerization of monosubstituted allenes in a stereoselective manner to give cross-conjugated trienes, which are different from those obtained by a palladium catalyst. PMID- 21647267 TI - Stereogenic boron in 2-amino-1,1-diphenylethanol-based boronate-imine and amine complexes. AB - Racemic boronate-imine and boronate-amine complexes 8 and 10, both featuring a stereogenic boron atom were synthesized from 2-amino-1,1-diphenylethanol (5) and characterized by crystal structure analyses. Proof of enantiomerism at the boron center for the novel boronate-amine complex 10 was established by separation of the enantiomers. Racemization barriers were found to be in the same range for both amine and imine complexes (100-110 kJ/mol). PMID- 21647268 TI - Identification of differentially expressed genes in uveal melanoma using suppressive subtractive hybridization. AB - PURPOSE: Uveal melanoma (UM) is the most common primary cancer of the eye, resulting not only in vision loss, but also in metastatic death. This study attempts to identify changes in the patterns of gene expression that lead to malignant transformation and proliferation of normal uveal melanocytes (UVM) using the Suppressive Subtractive Hybridization (SSH) technique. METHODS: The SSH technique was used to isolate genes that are differentially expressed in the TP31 cell line derived from a primary UM compared to UVM. The expression level of selected genes was further validated by microarray, semi-quantitative RT-PCR and western blot analyses. RESULTS: Analysis of the subtracted libraries revealed that 37 and 36 genes were, respectively, up- and downregulated in TP31 cells compared to UVM. Differential expression of the majority of these genes was confirmed by comparing UM cells with UVM by microarray. The expression pattern of selected genes was analyzed by semi-quantitative RT-PCR and western blot, and was found to be consistent with the SSH findings. CONCLUSIONS: We demonstrated that the SSH technique is efficient to detect differentially expressed genes in UM. The genes identified in this study represent valuable candidates for further functional analysis in UM and should be informative in studying the biology of this tumor. PMID- 21647269 TI - A novel mutation in the connexin 46 (GJA3) gene associated with congenital cataract in a Chinese pedigree. AB - PURPOSE: To identify the potential pathogenic mutation in a three-generation Chinese family with congenital nuclear pulverulent cataracts. METHODS: A three generation pedigree was recruited for our study. Three patients and four healthy members of the family underwent a comprehensive clinical examination. Genomic DNA extracted from peripheral blood was amplified using the polymerase chain reaction (PCR) method and the exons of all candidate genes were sequenced. RESULTS: When sequencing the encoding regions of the candidate genes, a novel mutation (c.559C>T) was identified in the gap junction protein alpha 3 (GJA3) gene, which resulted in the substitution of highly conserved proline by serine at codon 187 (P187S). There was no noticeable nucleotide polymorphism in other candidate genes. The mutation co-segregated with all patients, but was absent in the healthy members and 100 normal individuals. CONCLUSIONS: The present study identified a novel mutation (c.559C>T) in the GJA3 gene associated with autosomal dominant pulverulent cataracts in a Chinese family. As the first report to relate p.P187S mutation in GJA3, it expands the mutation spectrum of GJA3 in association with congenital cataracts. PMID- 21647270 TI - A novel mutation in the MIP gene is associated with autosomal dominant congenital nuclear cataract in a Chinese family. AB - PURPOSE: Congenital cataracts are a clinically and genetically heterogeneous lens disorder. The purpose of this study was to identify the genetic mutation and the molecular phenotype responsible for the presence of autosomal dominant congenital nuclear cataract disease in a Chinese family. METHODS: Patients were given physical examinations and their blood samples were collected for DNA extraction. Genotyping was performed by microsatellite markers and logarithm-of-odds (LOD) scores were calculated using the LINKAGE programs. Mutation detection was performed by direct sequencing. RESULTS: Linkage to the major intrinsic protein (MIP) locus was identified. Sequencing MIP revealed an A->G transition at nucleotide position c.530, which caused a conservative substitution of Tyr to Cys at codon 177 (P.Y177C). The Y177C mutation is located in the fifth transmembrane sequence. This mutation was identified in all affected individuals but is not found in any of the 100 control chromosomes. CONCLUSIONS: Our results identify that the c.530 (A->G) mutation in MIP is responsible for the Chinese pedigree. Our results further identify that the mutation in MIP is responsible for congenital cataract. The mutation found in our study broadens the spectrum of MIP mutations. PMID- 21647271 TI - The crucial role of collagen-binding integrins in maintaining the mechanical properties of human scleral fibroblasts-seeded collagen matrix. AB - PURPOSE: The aim of this study was to identify the presence of collagen-binding integrin subunits in human scleral fibroblasts (HSFs) and investigate their actual functions in maintaining the mechanical creep properties of the HSFs seeded collagen matrix. METHODS: Primary HSFs were cultured in vitro. Reverse- transcription PCR was used to detect mRNA expression of integrin alpha1, alpha2, and beta1 subunits in HSFs. In addition, western blot analysis and immunofluorescence were used to detect their protein in HSFs. Monoclonal antibodies were applied directly against the extracellular domains of integrin subunits in HSFs cultured in the three-dimensional collagen gels to block the interaction between HSFs and the extracellular collagen matrix. The effects of anti-integrin antibodies on HSFs morphology in collagen gel were observed. The effects of the added antibodies on fibroblast-mediated collagen gels' contraction were evaluated. Furthermore, the changes in mechanical creep properties of collagen gel were measured by a biomechanics test instrument. RESULTS: The mRNA and protein expressions of collagen-binding integrin alpha1, alpha2, and beta1 subunits were present in HSFs. The elongated bipolar cells converted to spherical shapes after 6 h after the addition of integrin alpha1beta1 and alpha2beta1 antibody. The blocking of integrin alpha1beta1 and alpha2beta1 subunits noticeably decreased the contraction in the collagen gels. In addition, all samples were subjected to a constantly applied load of 0.03 N for 600 s. The blocking of integrin alpha1beta1 and alpha2beta1 subunits also induced increases in the values of final extension, creep extension, and creep rate, compared to those of the controls (p<0.01). Furthermore, the creep elements were significantly increased with the augmentation of the integrin antibody dose (p<0.01). The final extension of the integrin alpha2beta1 antibody (1 MUg/ml or 4 MUg/ml) group was significantly higher compared to that of the integrin alpha1beta1 antibody (1 MUg/ml or 4 MUg/ml) group (p<0.01). However, the creep extension and creep rate of the integrin alpha2beta1 antibody (1 MUg/ml or 4 MUg/ml) group were not significantly different from those in the integrin alpha1beta1 antibody (1 MUg/ml or 4 MUg/ml) group (p>0.05). CONCLUSIONS: Our findings suggested that HSF integrin alpha1beta1 and alpha2beta1 participated in maintaining the mechanical creep properties of the HSFs-seeded collagen matrix. Furthermore, integrin alpha2beta1 might play a more crucial role in maintaining the mechanical creep properties of the collagen matrix than does integrin alpha1beta1. PMID- 21647272 TI - Current Status and Evidence Base of Training for Foster and Treatment Foster Parents. PMID- 21647273 TI - A GCG expansion (GCG)11 in polyadenylate-binding protein nuclear 1 gene caused oculopharyngeal muscular dystrophy in a Chinese family. AB - PURPOSE: To identify the mutation in polyadenylate-binding protein nuclear 1 gene (PABPN1, previously termed PABP2) in a Chinese family with autosomal, dominantly inherited oculopharyngeal muscular dystrophy (OPMD). METHODS: Clinical and ophthalmologic examinations were conducted on available living family members from three generations. Genomic DNA was extracted from peripheral blood leukocytes of every available family member, and the fragment flanking the (GCG)(n) of the PABPN1 gene was amplified by PCR. Mutations were screened by DNA sequencing. Photographs of deceased family members were examined for signs of OPMD. RESULTS: Clinical features of OPMD were found in all patients in generation II except the youngest sister, and no clinical manifestations were found in generation III. Mutation sequencing demonstrated that (GCG)6 in the wild PABPN1 gene was expanded to heterozygous (GCG)11 in all affected family members and in some but not all unaffected members. CONCLUSIONS: In a Chinese family with autosomal dominantly inherited OPMD, a heterozygous (GCG)11 expansion was identified in all affected family members and in several young unaffected members. PMID- 21647274 TI - Laboratory aid to the diagnosis and therapy of infection in the neonate. AB - Despite the advances in perinatal and neonatal care and use of newer potent antibiotics, the incidence of neonatal sepsis remains high and the outcome is still severe. For years, investigators have sought a test or panel of tests able to identify septic neonates accurately and rapidly in order to obtain an early diagnosis and develop a specific effective treatment for a successful outcome. In addition to the standard procedures (blood, CSF, and urine cultures), such panels have included a combination of haematological investigations (total, differential and immature cell counts), and levels of acute-phase reactants (principally CRP and procalcitonin), and cytokines (such as IL-6 or neutrophil CD64). Furthermore, the science of proteomics and genomics has been applied to the search for bio markers, production of protein profiles and genetic polymorphisms that can rapidly help the prediction, early diagnosis, and treatment of human diseases, but, for now, data are as yet insufficient to confirm their validity. PMID- 21647275 TI - Association of environment and place of birth with asthma in Chinese immigrant children. AB - Despite the advances in perinatal and neonatal care and use of newer potent antibiotics, the incidence of neonatal sepsis remains high and the outcome is still severe. For years, investigators have sought a test or panel of tests able to identify septic neonates accurately and rapidly in order to obtain an early diagnosis and develop a specific effective treatment for a successful outcome. In addition to the standard procedures (blood, CSF, and urine cultures, chest x ray), such panels have included a combination of total and differential cell counts, total immature neutrophil counts, immature to total neutrophil ratio, platelet counts, and levels of acute-phase reactants and cytokines. Furthermore, the science of proteomics and genomics has been applied to the search for biomarkers, production of protein profiles and genetic polymorphisms that can rapidly help the prediction, early diagnosis, and treatment of human diseases, but, for now, data are as yet insufficient to confirm their validity. PMID- 21647276 TI - Antibacterial prophylaxis in neutropenic children with cancer. AB - During the period of neutropenia induced by chemotherapy, patients have a high risk of infection. The use of antibiotic prophylaxis to reduce neutropenia related complications in patients with cancer is still disputed. Recent meta analysis and clinical trials demonstrated that antibiotic prophylaxis with quinolones reduces febrile episodes, bacterial infections and mortality in adult oncological patients with neutropenia induced by chemotherapy in acute leukaemia. In paediatric patients, the only randomized, double-blind, prospective study until now suggests that amoxicillin/clavulanate may represent an effective prophylactic treatment in reducing fever and infections in oncological children with neutropenia, with an efficacy that is statistically demonstrated only in patients with acute leukaemia. Considering the risk of resistances, antibiotic prophylaxis should be used only in selected patients. PMID- 21647277 TI - Consensus on diagnosis and empiric antibiotic therapy of febrile neutropenia. AB - Controversial issues on the management of empiric therapy and diagnosis of febrile neutropenia (FN) were faced by a Consensus Group of the Italian Association of Pediatric Hematology-Oncology (AIEOP). In this paper we report the suggestions of the consensus process regarding the role of aminoglycosides, glycopeptides and oral antibiotics in empiric therapy of FN, the rules for changing or discontinuing the therapy as well as the timing of the blood cultures. PMID- 21647278 TI - Prophylaxis and therapy of viral infections in pediatric patients treated for malignancy. AB - Infections are still an important cause of mortality and morbidity in pediatric cancer patients. Most of the febrile episodes in immunocompromised patients are classified as a fever of unknown origin (FUO) while bacteria are the more frequent causes of documented infections. Viral infections are also feared during chemotherapy but less data are available on their incidence and morbidity. We reviewed the literature on incidence, morbidity, and mortality of viral infections in children undergoing chemotherapy and discussed the evidence concerning the prophylaxis and the therapy. PMID- 21647279 TI - Current evidence of antifungal prophylaxis and therapy in pediatric patients. AB - Invasive fungal infections (IFI) are an important complication in pediatric haematological and oncological patients who undergo intensive chemotherapy for leukemia, solid tumour at advanced stage or relapsed, and hematopoietic stem cell transplantation. The incidence of IFI is lower than bacterial infection but mortality rate remains high. This review is designed to help paediatric oncologists in choosing the appropriate anti-fungal strategy and agents for prophylaxis, empirical, pre-emptive and specific therapy on the basis of published evidence. PMID- 21647280 TI - Guidelines for the management of bacterial and fungal infections during chemotherapy for pediatric acute leukemia or solid tumors: what is available in 2010? AB - Febrile episodes and infections represent important complications during antineoplastic chemotherapy for pediatric neoplastic diseases. In the last years many international association published guidelines for the management of these complications in adults, but no document of this type was prepared for children. One of the major causes of this situation is probably the very low number of pediatric clinical trials with adequate power and design. The paper summarizes guidelines provided for the management of infectious complications in adults with cancer by different international and will comment on how much they may be translated in the management of pediatric patients. PMID- 21647281 TI - Outpatient management of febrile neutropenia in children with cancer. AB - Optimizing the therapeutic strategies based on the results of randomized studies comparing different regimens led to a better prognosis of nearly all pediatric malignancies during the past four decades. Fever and neutropenia (FN) is a common complication in patients undergoing chemotherapy to treat cancer. There is no consensus on when standard therapy can be safely reduced; this lack of consensus leads to important variations in management of FN between different institutions, usually conducted according to local attitudes. To address this issue, the Infection working group of the Italian association for pediatric hematology oncology (AIEOP) organized a consensus meeting. This paper reports the agreement derived from this meeting. PMID- 21647282 TI - Preventing transmission of infectious agents in the pediatric in-patients hematology-oncology setting: what is the role for non-pharmacological prophylaxis? AB - The most intensive chemotherapy regimens were used in the past for leukemia patients who were the main focus of trials on infections; today there are increasing numbers of children with solid cancer and considerable risk of infection who do receive intensive standard-dose chemotherapy. Despite a continuous will to protect the immune-compromised child from infections, evidence based indications for intervention by non-pharmacological tools is still lacking in the pediatric hematology-oncology literature. Guidelines on standard precautions as well as precautions to avoid transmission of specific infectious agents are available. As a result of a consensus discussion, the Italian Association for Pediatric Hematology-Oncology (AIEOP) Cooperative Group centers agree that for children treated with chemotherapy both of these approaches should be implemented and vigorously enforced, while additional policies, including strict environmental isolation, should be restricted to patients with selected clinical conditions or complications. We present here a study by the working group on infectious diseases of AIEOP. PMID- 21647283 TI - Spectrophotometric determination of N-acetyl-L-cysteine and N-(2 mercaptopropionyl)-glycine in pharmaceutical preparations. AB - A simple spectrophotometric method for the determination of N-acetyl-L-cysteine (NAC) and N-(2-mercaptopropionyl)glycine (MPG) in pharmaceutical preparations was developed, validated, and used. The proposed equilibrium method is based on a coupled two-step redox and complexation reaction. In the first step, Fe(III) is reduced to Fe(II) by NAC or MPG. Subsequently, Fe(II) is complexed with 2,4,6 tripyridyl-s-triazine (TPTZ). Several analytical parameters of the method were optimized for NAC and MPG analysis in the concentration range from 1.0 MUM to 100.0 MUM. Regression analysis of the calibration data showed a good correlation coefficient (0.9999). The detection limit of the method was 0.14 MUM for NAC and 0.13 MUM for MPG. The method was successfully applied to quantify NAC and MPG in pharmaceutical preparations. No interferences were observed from common pharmaceutical excipients. PMID- 21647284 TI - Comparison of two methods for assaying reducing sugars in the determination of carbohydrase activities. AB - The Nelson-Somogyi (NS) and 3,5-dinitrosalicylic acid (DNS) assays for reducing sugars are widely used in measurements of carbohydrase activities against different polysaccharides. Using twelve commercial enzyme preparations, the comparison of the NS and DNS assays in determination of cellulase, beta glucanase, xylanase, and beta-mannanase activities was carried out. When cellulase activities against CMC were measured, the DNS assay gave activity values, which were typically 40-50% higher than those obtained with the NS assay. In the analysis of the xylanase, beta-mannanase, and beta-glucanase activities, the overestimations by the DNS assay were much more pronounced (the observed differences in the activities were 3- to 13-fold). Reasons for preferential use of the NS assay for measuring activities of carbohydrases other than cellulases are discussed. PMID- 21647285 TI - Determination of diethyl phthalate and polyhexamethylene guanidine in surrogate alcohol from Russia. AB - Analytical methods based on spectroscopic techniques were developed and validated for the determination of diethyl phthalate (DEP) and polyhexamethylene guanidine (PHMG), which may occur in unrecorded alcohol. Analysis for PHMG was based on UV VIS spectrophotometry after derivatization with Eosin Y and (1)H NMR spectroscopy of the DMSO extract. Analysis of DEP was performed with direct UV-VIS and (1)H NMR methods. Multivariate curve resolution and spectra computation methods were used to confirm the presence of PHMG and DEP in the investigated beverages. Of 22 analysed alcohol samples, two contained DEP or PHMG. (1)H NMR analysis also revealed the presence of signals of hawthorn extract in three medicinal alcohols used as surrogate alcohol. The simple and cheap UV-VIS methods can be used for rapid screening of surrogate alcohol samples for impurities, while (1)H NMR is recommended for specific confirmatory analysis if required. PMID- 21647286 TI - A Comparative Electrochemical Behaviour Study and Analytical Detection of the p Nitrophenol Using Silver Solid Amalgam, Mercury, and Silver Electrodes. AB - This work reports a comparative electrochemical behaviour study and p-nitrophenol analytical detection using silver solid amalgam, hanging dropping mercury, and silver electrodes. For this, square wave voltammetry was employed, where the analytical responses and the redox mechanisms could be compared for reduction processes of 4-nitrophenol by analysis of the voltammetric responses. The analytical performance of the electrode was evaluated and detection and quantification limits, recovery percentages, repeatability, and reproducibility for the silver solid amalgam and hanging dropping mercury electrodes presented similar values; the results presented for the silver electrode indicated worse analytical parameters than the other electrodes. The results indicate that the silver solid amalgam electrode can be considered a suitable tool and an interesting alternative for the analytical determination of 4-nitrophenol, as well as for the determination of other biological and environmentally interesting compounds that present analytical responses on mercury surfaces. PMID- 21647287 TI - A new spectrophotometric method for determination of selenium in cosmetic and pharmaceutical preparations after preconcentration with cloud point extraction. AB - A simple, rapid, and sensitive spectrophotometric method for the determination of trace amounts of selenium (IV) was described. In this method, all selenium spices reduced to selenium (IV) using 6 M HCl. Cloud point extraction was applied as a preconcentration method for spectrophotometric determination of selenium (IV) in aqueous solution. The proposed method is based on the complexation of Selenium (IV) with dithizone at pH < 1 in micellar medium (Triton X-100). After complexation with dithizone, the analyte was quantitatively extracted to the surfactant-rich phase by centrifugation and diluted to 5 mL with methanol. Since the absorption maxima of the complex (424 nm) and dithizone (434 nm) overlap, hence, the corrected absorbance, Acorr, was used to overcome the problem. With regard to the preconcentration, the tested parameters were the pH of the extraction, the concentration of the surfactant, the concentration of dithizone, and equilibration temperature and time. The detection limit is 4.4 ng mL(-1); the relative standard deviation for six replicate measurements is 2.18% for 50 ng mL( 1) of selenium. The procedure was applied successfully to the determination of selenium in two kinds of pharmaceutical samples. PMID- 21647288 TI - Fluorescence Spectrometric Determination of Drugs Containing alpha-Methylene Sulfone/Sulfonamide Functional Groups Using N-Methylnicotinamide Chloride as a Fluorogenic Agent. AB - A simple spectrofluorometric method has been developed, adapted, and validated for the quantitative estimation of drugs containing alpha-methylene sulfone/sulfonamide functional groups using N(1)-methylnicotinamide chloride (NMNCl) as fluorogenic agent. The proposed method has been applied successfully to the determination of methyl sulfonyl methane (MSM) (1), tinidazole (2), rofecoxib (3), and nimesulide (4) in pure forms, laboratory-prepared mixtures, pharmaceutical dosage forms, spiked human plasma samples, and in volunteer's blood. The method showed linearity over concentration ranging from 1 to 150 MUg/mL, 10 to 1000 ng/mL, 1 to 1800 ng/mL, and 30 to 2100 ng/mL for standard solutions of 1, 2, 3, and 4, respectively, and over concentration ranging from 5 to 150 MUg/mL, 10 to 1000 ng/mL, 10 to 1700 ng/mL, and 30 to 2350 ng/mL in spiked human plasma samples of 1, 2, 3, and 4, respectively. The method showed good accuracy, specificity, and precision in both laboratory-prepared mixtures and in spiked human plasma samples. The proposed method is simple, does not need sophisticated instruments, and is suitable for quality control application, bioavailability, and bioequivalency studies. Besides, its detection limits are comparable to other sophisticated chromatographic methods. PMID- 21647289 TI - Evaluation of acetylcholinesterase biosensor based on carbon nanotube paste in the determination of chlorphenvinphos. AB - An amperometric biosensor for chlorphenvinphos (organophosphorus pesticide) based on carbon nanotube paste and acetylcholinesterase enzyme (CNTs-AChE biosensor) is described herein. This CNTs-AChE biosensor was characterized by scanning electron microscopy (SEM) and electrochemical impedance spectroscopy (EIS). The SEM result shows the presence of CNTs and small lumps, due to the enzyme AChE, which has a type of cauliflower formation. From EIS analysis is possible to observe increased R(tc) for CNTs-AChE biosensor when compared to the carbon nanotube paste electrode for the reaction [Fe(CN)(6)](4-/3-). Using a chronoamperometric procedure, a linear analytical curve was observed in the 4.90 * 10(-7)-7.46 * 10( 6) M range with limit of detection of 1.15 * 10(-7) M. The determination of chlorphenvinphos in the insecticide sample proved to be in agreement with the standard spectrophotometric method, with a 95% confidence level and with a relative error lower than 3%. In this way, the CNTs-AChE biosensor presented easy preparation, fast response, sensitivity, durability, good repeatability, and reproducibility. PMID- 21647290 TI - Day and Night GSH and MDA Levels in Healthy Adults and Effects of Different Doses of Melatonin on These Parameters. AB - The pineal secretory product melatonin (chemically, N-acetyl-5-methoxytryptamine) acts as an effective antioxidant and free-radical scavenger and plays an important role in several physiological functions such as sleep induction, immunomodulation, cardiovascular protection, thermoregulation, neuroprotection, tumor-suppression and oncostasis. Membrane lipid-peroxidation in terms of malondialdehyde (MDA) and intracellular glutathione (GSH) is considered to be a reliable marker of oxidative stress. The present work was undertaken to study the modulating effect of melatonin on MDA and GSH in human erythrocytes during day and night. Our observation shows the modulation of these two biomarkers by melatonin, and this may have important therapeutic implications. In vitro dose dependent effect of melatonin also showed variation during day and night. We explain our observations on the basis of melatonin's antioxidative function and its effect on the fluidity of plasma membrane of red blood cells. Rhythmic modulation of MDA and GSH contents emphasized the role of melatonin as an antioxidant and its function against oxidative stress. PMID- 21647291 TI - The mammary gland microenvironment directs progenitor cell fate in vivo. AB - The mammary gland is a unique organ that continually undergoes postnatal developmental changes. In mice, the mammary gland is formed via signals from terminal end buds, which direct ductal growth and elongation. Intriguingly, it is likely that the entire cellular repertoire of the mammary gland is formed from a single antecedent cell. Furthermore, in order to produce progeny of varied lineages (e.g., luminal and myoepithelial cells), signals from the local tissue microenvironment influence mammary stem/progenitor cell fate. Data have shown that cells from the mammary gland microenvironment reprogram adult somatic cells from other organs (testes, nerve) into cells that produce milk and express mammary epithelial cell proteins. Similar results were found for human tumorigenic epithelial carcinoma cells. Presently, it is unclear how the deterministic power of the mammary gland microenvironment controls epithelial cell fate. Regardless, signals generated by the microenvironment have a profound influence on progenitor cell differentiation in vivo. PMID- 21647292 TI - Effector caspases and leukemia. AB - Caspases, a family of aspartate-specific cysteine proteases, play a major role in apoptosis and a variety of physiological and pathological processes. Fourteen mammalian caspases have been identified and can be divided into two groups: inflammatory caspases and apoptotic caspases. Based on the structure and function, the apoptotic caspases are further grouped into initiator/apical caspases (caspase-2, -8, -9, and -10) and effector/executioner caspases (caspase 3, -6, and -7). In this paper, we discuss what we have learned about the role of individual effector caspase in mediating both apoptotic and nonapoptotic events, with special emphasis on leukemia-specific oncoproteins in relation to effector caspases. PMID- 21647293 TI - Investigating the purpose of prelamin A processing. AB - Lmna yields two major protein products in somatic cells, lamin C and prelamin A. Mature lamin A is produced from prelamin A by four posttranslational processing steps-farnesylation of a carboxyl-terminal cysteine, release of the last three amino acids of the protein, methylation of the farnesylcysteine, and the endoproteolytic release of the carboxyl-terminal 15 amino acids of the protein (including the farnesylcysteine methyl ester). Although the posttranslational processing of prelamin A has been conserved in vertebrate evolution, its physiologic significance remains unclear. Here we review recent studies in which we investigated prelamin A processing with Lmna knock-in mice that produce exclusively prelamin A (Lmna(PLAO)), mature lamin A (Lmna(LAO)) or nonfarnesylated prelamin A (Lmna(nPLAO)). We found that the synthesis of lamin C is dispensable in laboratory mice, that the direct production of mature lamin A (completely bypassing all prelamin A processing) causes no discernable pathology in mice, and that exclusive production of nonfarnesylated prelamin A leads to cardiomyopathy. PMID- 21647294 TI - 'Natively unfolded' nucleoporins in nucleocytoplasmic transport: clustered or evenly distributed? AB - The nuclear pore complex (NPC) acts as a selective gate that mediates the bidirectional transport of macromolecules between the cytoplasm and the nucleus of eukaryotic cells. 'Natively unfolded' nucleoporins (Nups) with domains rich in phenylalanine-glycine (FG) repeats form the selective permeability barrier and provide binding sites for mobile transport receptors in the NPC. Understanding the structure and function of the FG-Nups barrier under real-time trafficking conditions is still a formidable challenge due to the dynamic nature of a channeled membranous environment. Recently, we have shown that three-dimensional (3D) density maps of transient interactions between the FG-Nups barrier and a cargo-free or a cargo-bound transport receptor in native NPCs can be obtained by an advanced single-molecule fluorescence microscopy approach. Moreover, we found that these interaction sites are spatially clustered into distinct groups in the periphery around a central axial channel with a diameter of approximately 10-20 nm in the NPC. The 3D distribution of interaction sites may indicate some native properties of the FG-Nups barrier. Here we speculate that the selective permeability barrier in the NPC could be formed by clustered FG-Nups. PMID- 21647295 TI - Determining nuclear shape: the role of farnesylated nuclear membrane proteins. AB - Changes in nuclear morphology are observed in diverse developmental processes as well as in pathological conditions. Modification of nuclear membrane and nuclear lamina protein levels results in altered nuclear shapes, as it has been demonstrated in experimental systems ranging from yeast to human cells. The important role of nuclear membrane components in regulating nuclear morphology is additionally highlighted by the abnormally shaped nuclei observed in diseases where nuclear lamina proteins are mutated. Even though the effect of nuclear envelope components on nuclear shape has been thoroughly described, not much is known about the molecular mechanisms that govern these events. In addition to the known role of intermediate filament formation by lamins, here we discuss several mechanisms that might alone or in combination participate in the regulation of nuclear shape observed upon modification of the levels of nuclear membrane and lamina proteins. Based on recent work with the two farnesylated nuclear membrane Drosophila proteins, kugelkern and lamin Dm0, we propose that the direct interaction of farnesylated nuclear membrane proteins with the phospholipid bilayer leads to nuclear envelope deformation. In addition to this mechanism, we suggest that the interaction of nuclear membrane and lamina proteins with cytoskeletal elements and chromatin, and modifications in lipid biosynthesis might also be involved in the formation of abnormally shaped nuclei. PMID- 21647296 TI - Telomere dysfunction in human bone marrow failure syndromes. AB - Approximately 90% of all human cancers, in which some deregulation of cell cycle arrest or programmed cell death has occurred, express telomerase, a ribonucleoprotein whose activity is normally turned off in healthy somatic tissues. Additionally, small populations of self-renewing stem cells, such as hematopoietic stem cells, skin and hair follicle basal layer cells and intestinal basal crypt cells, have been shown to retain telomerase activity. Conversely, hereditary defects that result in shortened telomeres in humans have been shown to manifest most often as bone marrow failure or pulmonary fibrosis, along with a myriad of other symptoms, likely due to the loss of the stem and/or progenitor cells of affected tissues. The aim of this review is to highlight our knowledge of the mechanisms of telomere maintenance that contribute to the pathology of human disease caused by dysfunctional telomere homeostasis. Specifically, a new role for the SNM1B/Apollo nuclease in the pathologies of Hoyeraal-Hreidarsson syndrome will be discussed. PMID- 21647297 TI - A role for SUMOylation in snoRNP biogenesis revealed by quantitative proteomics. AB - A role for SUMOylation in the biogenesis and/or function of Box C/D snoRNPs has been reported, mediated via SUMO2 conjugation to the core snoRNP protein, Nop58. A quantitative proteomics screen, based on SILAC (stable-isotope labeling by amino acids in cell culture) and mass spectrometry using extracts prepared from cultured mammalian cells expressing either 6His-SUMO1 or -SUMO2, revealed that the snoRNP-related proteins Nop58, Nhp2, DKC1 and NOLC1 are amongst the main SUMO modified proteins in the nucleolus. SUMOylation of Nhp2 and endogenous Nop58 was confirmed using a combination of in vitro and cell-based assays and the modified lysines identified by site-directed mutagenesis. SUMOylation of Nop58 was found to be important for high-affinity Box C/D snoRNA binding and the localization of newly transcribed snoRNAs to the nucleolus. Here, these findings are reviewed and a model for understanding Nop58 SUMOylation in the context of Box C/D snoRNP biogenesis is presented. Given the essential role of snoRNPs in the modification of pre-ribosomal RNA, this work suggests that SUMO, snoRNPs and ribosome assembly, and thus cellular translation, growth and proliferation, may be linked via novel mechanisms which warrant further investigation. PMID- 21647298 TI - Actin-related proteins localized in the nucleus: from discovery to novel roles in nuclear organization. AB - The actin family consists of conventional actin and actin-related proteins (ARPs), and the members show moderate similarity and share the same basal structure. Following the finding of various ARPs in the cytoplasm in the 1990s, multiple subfamilies that are localized predominantly in the nucleus were identified. Consistent with these cytological observations, subsequent biochemical analyses revealed the involvement of the nuclear ARPs in ATP dependent chromatin-remodeling and histone acetyltransferase complexes. In addition to their contribution to chromatin remodeling, recent studies have shown that nuclear ARPs have roles in the organization of the nucleus that are independent of the activity of the above-mentioned complexes. Therefore, nuclear ARPs are recognized as novel key regulators of genome function, and affect not only the remodeling of chromatin but also the spatial arrangement and dynamics of chromatin within the nucleus. PMID- 21647299 TI - An epichromatin epitope: persistence in the cell cycle and conservation in evolution. AB - Interphase nuclear architecture is disrupted and rapidly reformed with each cell division cycle. Successive cell generations exhibit a "memory" of this nuclear architecture, as well as for gene expression. Furthermore, many features of nuclear and mitotic chromosome structure are recognizably species and tissue specific. We wish to know what properties of the underlying chromatin structure may determine these conserved features of nuclear architecture. Employing a particular mouse autoimmune anti-nucleosome monoclonal antibody (PL2-6), combined with deconvolution immunofluorescence microscopy, we present evidence for a unique epitope (involving a ternary complex of histones H2A and H2B and DNA) which is localized only at the exterior chromatin surface of interphase nuclei and mitotic chromosomes in mammalian, invertebrate and plant systems. As only the surface chromatin region is identified with antibody PL2-6, we have assigned it the name "epichromatin". We describe an "epichromatin hypothesis", suggesting that epichromatin may have a unique evolutionary conserved conformation which facilitates interaction with the reforming post-mitotic nuclear envelope and a rapid return of interphase nuclear architecture. PMID- 21647300 TI - Histone deacetylase inhibition redistributes topoisomerase IIbeta from heterochromatin to euchromatin. AB - The genome is organized into large scale structures in the interphase nucleus. Pericentromeric heterochromatin represents one such compartment characterized by histones H3 and H4 tri-methylated at K9 and K20 respectively and with a correspondingly low level of histone acetylation. HP1 proteins are concentrated in pericentric heterochromatin and histone deacetylase inhibitors such as trichostatin A (TSA) promote hyperacetylation of heterochromatic nucleosomes and the dispersal of HP1 proteins. We observed that in mouse cells, which contain prominent heterochromatin, DNA topoisomerase IIbeta (topoIIbeta) is also concentrated in heterochromatic regions. Similarly, a detergent-resistant fraction of topoIIbeta is associated with heterochromatin in human cell lines. Treatment with TSA displaced topoIIbeta from the heterochromatin with similar kinetics to the displacement of HP1beta. Topoisomerase II is the cellular target for a number of clinically important cytotoxic anti-cancer agents known collectively as topoisomerase poisons, and it has been previously reported that histone deacetylase inhibitors can sensitize cells to these drugs. While topoIIalpha appears to be the major target for most topoisomerase poisons, histone deacetylase-mediated potentiation of these drugs is dependent on topoIIbeta. We find that while prior treatment with TSA did not increase the quantity of etoposide-mediated topoIIbeta-DNA covalent complexes, it did result in a shift in their distribution from a largely heterochromatin-associated to a pannuclear pattern. We suggest that this redistribution of topoIIbeta converts this isoform of topoII to a effective relevant target for topoisomerase poisons. PMID- 21647301 TI - The F-actin severing protein cofilin-1 is required for RNA polymerase II transcription elongation. AB - In mammals actin contributes to transcription elongation by facilitating establishment of permissive chromatin. Here we report that the F-actin severing protein cofilin-1 is part of the same complex with actin and phosphorylated RNA polymerase (pol) II. In chromatin immunoprecipitation assays cofilin-1 was found selectively associated with transcribed regions of active genes, its occupancy being influenced by the polymerization state of actin. Cofilin-1 gene silencing led to a drop in FUrd incorporation into nascent transcripts. In cofilin-1 silenced cells chromatin immunoprecipitations showed that active genes were devoid of actin, phosphorylated pol II and displayed low histone H3 acetylation levels on K9. These findings suggest that cofilin-1 plays a major role in pol II transcription, facilitating association of elongating pol II and actin with active genes. We speculate that cofilin-1 performs its function in pol II transcription by regulating polymerization of gene-associated actin. PMID- 21647302 TI - The DNA binding and bending activities of truncated tail-less HMGB1 protein are differentially affected by Lys-2 and Lys-81 residues and their acetylation. AB - The role of lysines 2 and 81 as target sites for acetylation in full-length HMGB1 and truncated tail-less protein, respectively, has been studied by mutation analysis for the abilities of these proteins to bind and bend DNA. The DNA bending ability of truncated tail-less HMGB1 containing Lys-2 mutated to alanine does not differ from that of the wild-type protein, while the same mutation of Lys-81 reduced the bending capacity of the mutant protein. These data demonstrate that Lys-81 is critical for the DNA bending ability of truncated HMGB1. Such a conclusion is further confirmed by the experiments carried out with CBP acetylated proteins: acetylation of Lys-2 in mutant protein K81/A81 alleviated DNA bending and induced DNA end-joining. On the contrary, the acetylation of Lys 81 in the mutant K2/A2 enhanced the bending potential of HMGB1?C. Regarding the ability of HMGB1 to specifically bind bent DNA, the individual mutations of either K2 or K81 as well as the double mutation of both residues to alanine were found to completely abolish binding of truncated tail-less HMGB1 to cisplatin modified DNA. We conclude that unlike the case with the bending ability of truncated HMGB1, where Lys-81 has a primary function, Lys-2 and Lys-81 are both critical for the protein's binding to cisplatin-modified DNA. The mutation K2/A2 in full-length HMGB1 and acidic tail removal induce the same conformational changes. Any further substitutions at the acetylable lysines in the truncated form of HMGB1 do not have an additional effect. PMID- 21647303 TI - Modeling the effects of a simple immune system and immunodeficiency on the dynamics of conjointly growing tumor and normal cells. AB - In this paper, we develop a theoretical contribution towards the understanding of the complex behavior of conjoint tumor-normal cell growth under the influence of immuno-chemotherapeutic agents under simple immune system response. In particular, we consider a core model for the interaction of tumor cells with the surrounding normal cells. We then add the effects of a simple immune system, and both immune-suppression factors and immuno-chemotherapeutic agents as well. Through a series of numerical simulations, we illustrate that the interdependency of tumor-normal cells, together with choice of drug and the nature of the immunodeficiency, leads to a variety of interesting patterns in the evolution of both the tumor and the normal cell populations. PMID- 21647304 TI - Difference of sodium currents between pediatric and adult human atrial myocytes: evidence for developmental changes of sodium channels. AB - Voltage-gated calcium currents and potassium currents were shown to undergo developmental changes in postnatal human and animal cardiomocytes. However, so far, there is no evidence whether sodium currents also presented the developmental changes in postnatal human atrial cells. The aim of this study was to observe age-related changes of sodium currents between pediatric and adult atrial myocytes. Human atrial myocytes were acutely isolated and the whole-cell patch clamp technique was used to record sodium currents isolated from pediatric and adult atrial cardiomocytes. The peak amplitude of sodium currents recorded in adult atrial cells was significantly larger than that in pediatric atrial myocytes. However, there was no significant difference of the activation voltage for peak sodium currents between two kinds of atrial myocytes. The time constants for the activation and inactivation of sodium currents were smaller in adult atria than pediatric atria. The further study revealed that the voltage-dependent inactivation of sodium currents were more slow in adult atrial cardiomyocytes than pediatric atrial cells. A significant difference was also observed in the recovery process of sodium channel from inactivation. In summary, a few significant differences were demonstrated in sodium currents characteristics between pediatric and adult atrial myocytes, which indicates that sodium currents in human atria also undergo developmental changes. PMID- 21647306 TI - Rational decision-making in inhibitory control. AB - An important aspect of cognitive flexibility is inhibitory control, the ability to dynamically modify or cancel planned actions in response to changes in the sensory environment or task demands. We formulate a probabilistic, rational decision-making framework for inhibitory control in the stop signal paradigm. Our model posits that subjects maintain a Bayes-optimal, continually updated representation of sensory inputs, and repeatedly assess the relative value of stopping and going on a fine temporal scale, in order to make an optimal decision on when and whether to go on each trial. We further posit that they implement this continual evaluation with respect to a global objective function capturing the various reward and penalties associated with different behavioral outcomes, such as speed and accuracy, or the relative costs of stop errors and go errors. We demonstrate that our rational decision-making model naturally gives rise to basic behavioral characteristics consistently observed for this paradigm, as well as more subtle effects due to contextual factors such as reward contingencies or motivational factors. Furthermore, we show that the classical race model can be seen as a computationally simpler, perhaps neurally plausible, approximation to optimal decision-making. This conceptual link allows us to predict how the parameters of the race model, such as the stopping latency, should change with task parameters and individual experiences/ability. PMID- 21647305 TI - Age sensitivity of behavioral tests and brain substrates of normal aging in mice. AB - Knowledge of age sensitivity, the capacity of a behavioral test to reliably detect age-related changes, has utility in the design of experiments to elucidate processes of normal aging. We review the application of these tests in studies of normal aging and compare and contrast the age sensitivity of the Barnes maze, eyeblink classical conditioning, fear conditioning, Morris water maze, and rotorod. These tests have all been implemented to assess normal age-related changes in learning and memory in rodents, which generalize in many cases to age related changes in learning and memory in all mammals, including humans. Behavioral assessments are a valuable means to measure functional outcomes of neuroscientific studies of aging. Highlighted in this review are the attributes and limitations of these measures in mice in the context of age sensitivity and processes of brain aging. Attributes of these tests include reliability and validity as assessments of learning and memory, well-defined neural substrates, and sensitivity to neural and pharmacological manipulations and disruptions. These tests engage the hippocampus and/or the cerebellum, two structures centrally involved in learning and memory that undergo functional and anatomical changes in normal aging. A test that is less well represented in studies of normal aging, the context pre-exposure facilitation effect (CPFE) in fear conditioning, is described as a method to increase sensitivity of contextual fear conditioning to changes in the hippocampus. Recommendations for increasing the age sensitivity of all measures of normal aging in mice are included, as well as a discussion of the potential of the under-studied CPFE to advance understanding of subtle hippocampus-mediated phenomena. PMID- 21647307 TI - Connexin 43 is a potential prognostic biomarker for ewing sarcoma/primitive neuroectodermal tumor. AB - Connexins (Cxs) are building unit proteins of gap junctions (GJs) that are prognostic markers in carcinomas. To investigate the role of Cx in Ewing sarcoma (EWS)/primitive neuroectodermal tumor (PNET), we examined the expression of Cx43 and Cx26 in 36 EWS/PNETs and found (1) cytoplasmic Cx43 reactivity in 28/36 (78%) cases. (2) Cx43 score was significantly correlated with overall survival (P = .025). The average scores for patients alive and dead at 3 years are 46.08 and 96.98 (P = .004) at 5 years are 46.06 and 96.42 (P = .002). (3) Metastasis had a significant effect on the overall survival (P = .003). (4) Cytoplasmic Cx26 reactivity was detected in 2 of 36 (6%) patients who died with metastasis. Our results suggest a possible oncogenic and prognostic role for Cx43 and Cx26 in EWS/PNET. The lack of membranous immunoreactivity suggests that the effect of Cx in EWS/PNET is via a GJ function-independent mechanism. PMID- 21647308 TI - Using web search query data to monitor dengue epidemics: a new model for neglected tropical disease surveillance. AB - BACKGROUND: A variety of obstacles including bureaucracy and lack of resources have interfered with timely detection and reporting of dengue cases in many endemic countries. Surveillance efforts have turned to modern data sources, such as Internet search queries, which have been shown to be effective for monitoring influenza-like illnesses. However, few have evaluated the utility of web search query data for other diseases, especially those of high morbidity and mortality or where a vaccine may not exist. In this study, we aimed to assess whether web search queries are a viable data source for the early detection and monitoring of dengue epidemics. METHODOLOGY/PRINCIPAL FINDINGS: Bolivia, Brazil, India, Indonesia and Singapore were chosen for analysis based on available data and adequate search volume. For each country, a univariate linear model was then built by fitting a time series of the fraction of Google search query volume for specific dengue-related queries from that country against a time series of official dengue case counts for a time-frame within 2003-2010. The specific combination of queries used was chosen to maximize model fit. Spurious spikes in the data were also removed prior to model fitting. The final models, fit using a training subset of the data, were cross-validated against both the overall dataset and a holdout subset of the data. All models were found to fit the data quite well, with validation correlations ranging from 0.82 to 0.99. CONCLUSIONS/SIGNIFICANCE: Web search query data were found to be capable of tracking dengue activity in Bolivia, Brazil, India, Indonesia and Singapore. Whereas traditional dengue data from official sources are often not available until after some substantial delay, web search query data are available in near real-time. These data represent valuable complement to assist with traditional dengue surveillance. PMID- 21647309 TI - A new approach to monitoring dengue activity. PMID- 21647310 TI - Once-daily tacrolimus extended-release formulation: 1 year after conversion in stable pediatric kidney transplant recipients. AB - It is speculated that a once-daily dosage of immunosuppression can increase adherence and thereby graft survival. Until now, there have been no studies on once-daily use of Tacrolimus extended-release formulation (TAC-ER) in children following pediatric kidney transplantation. In 11 stable pediatric kidney recipients >10 years, efficacy, safety, and tolerability of a switch to TAC-ER were observed over one year. Adherence was determined by use of the BAASIS-Scale Interview and comparison of individual variability of Tacrolimus trough levels. Over the observation period, two acute rejections were observed in one girl with nonadherence and repeated Tacrolimus trough levels of 0 ng/m. Beside this, there were no acute rejections in this trial. TAC dose was increased in 3/11 patients and decreased in 2/11 patients within the course of the study. Six patients did not require a dose adjustment. All but one patient had a maximum of 1 dose change during therapy. Mean Tacrolimus dose, trough levels, and Glomerular filtration rates were also stable. Adherence, as measured by BAASIS-Scale Interview and coefficient of variation of Tacrolimus trough levels, was good at all times. It is concluded that conversion to Tac-ER is safe in low-risk children following pediatric kidney transplantation. PMID- 21647311 TI - Hepatoprotective Effects of Orthosiphon stamineus Extract on Thioacetamide Induced Liver Cirrhosis in Rats. AB - Orthosiphon stamineus as medicinal plant is commonly used in Malaysia for treatment of hepatitis and jaundice; in this study, the ethanol extracts were applied to evaluate the hepatoprotective effects in a thioacetamide-induced hepatotoxic model in Sprague Dawley rats. Five groups of adult rats were arranged as follows: Group 1 (normal control group), Group 2 Thioacetamide (TAA) as positive control (hepatotoxic group), Group 3 Silymarin as a well-known standard drug (hepatoprotective group), and Groups 4 and 5 as high and low dose (treatment groups). After 60-day treatment, all rats were sacrificed. The hepatotoxic group showed a coarse granulation on the liver surface when compared to the smooth aspect observed on the liver surface of the other groups. Histopathological study confirmed the result; moreover, there was a significant increase in serum liver biochemical parameters (ALT, AST, ALP, and Bilirubin) and the level of liver Malondialdehyde (MDA), accompanied by a significant decrease in the level of total protein and Albumin in the TAA control group when compared with that of the normal group. The high-dose treatment group (200 mg/kg) significantly restored the elevated liver function enzymes near to normal. This study revealed that 200 mg/kg extracts of O. stamineus exerted a hepatoprotective effect. PMID- 21647312 TI - Resident stem cells and renal carcinoma. AB - According to the cancer stem cell hypothesis tumors are maintained by a cancer stem cell population which is able to initiate and maintain tumors. Tumor initiating stem cells display stem or progenitor cell properties such as self renewal and capacity to re-establish tumors that recapitulate the tumor of origin. In this paper, we discuss data relative to the presence of cancer stem cells in human renal carcinoma and their possible origin from normal resident stem cells. The cancer stem cells identified in human renal carcinomas are not derived from the normal CD133(+) progenitors of the kidney, but rather from a more undifferentiated population that retains a mesenchymal phenotype. This population is able to self-renewal, clonogenicity, and in vivo tumor initiation. Moreover, they retain pluripotent differentiation capability, as they can generate not only the epithelial component of the tumor, but also tumor endothelial cells. This suggests that renal cancer stem cells may contribute to the intratumor vasculogenesis. PMID- 21647313 TI - Resequencing DCDC5 in the Flanking Region of an LD-SNP Derived from a Kidney-Yang Deficiency Syndrome Family. AB - Objective. To explore the genetic traits of Kidney-yang deficiency syndrome (KDS). Design. Twelve KDS subjects and three spouses from a typical KDS family were recruited. Their genomic DNA samples were genotyped by Affymetrix 100K single-nucleotide polymorphism (SNP) arrays. The linkage disequilibrium (LD) SNPs were generated using GeneChip DNA analysis software (GDAS, Affymetrix). Genes located within 100 bp of the flanks of LD SNPs were mined via GeneView. 29 exons of the doublecortin domain containing 5 (DCDC5), a representative gene within the flank of an LD SNP, were resequenced. Results. Five LD SNPs display midrange linkage with KDS. Two genes with established functions, DCDC5 and Leucyl-tRNA synthetase, were mined in the flanks of LD SNPs. Resequencing of DCDC5 revealed a nonsynonymous variation, in which 3764T/A was replaced by C/G. Accordingly, the Ser(1172) was substituted by Pro(1172). The S1172P substitution effect was evaluated as "possibly damaging" by PolyPhen. Conclusion. We have identified a genomic variation of DCDC5 based on the LD SNPs derived from a KDS family. DCDC5 and other genes surrounding these SNPs display some relationships with key symptoms of KDS. PMID- 21647314 TI - Metabolic Profile of Offspring from Diabetic Wistar Rats Treated with Mentha piperita (Peppermint). AB - This study aimed at evaluating glycemia and lipid profile of offspring from diabetic Wistar rats treated with Mentha piperita (peppermint) juice. Male offspring from nondiabetic dams (control group: 10 animals treated with water and 10 treated with peppermint juice) and from dams with streptozotocin-induced severe diabetes (diabetic group: 10 animals treated with water and 10 treated with peppermint juice) were used. They were treated during 30 days, and, after the treatment period, levels of glycemia, triglycerides, total cholesterol, and fractions were analyzed in the adult phase. The offspring from diabetic dams treated with peppermint showed significantly reduced levels of glucose, cholesterol, LDL-c, and triglycerides and significant increase in HDL-c levels. The use of the M. piperita juice has potential as culturally appropriate strategy to aid in the prevention of DM, dyslipidemia, and its complications. PMID- 21647315 TI - Synergistic antibacterial effects of polyphenolic compounds from olive mill wastewater. AB - Polyphenols or phenolic compounds are groups of secondary metabolites widely distributed in plants and found in olive mill wastewater (OMW). Phenolic compounds as well as OMW extracts were evaluated in vitro for their antimicrobial activity against Gram-positive (Streptococcus pyogenes and Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli and Klebsiella pneumoniae). Most of the tested phenols were not effective against the four bacterial strains when tested as single compounds at concentrations of up to 1000 MUg mL(-1). Hydroxytyrosol at 400 MUg mL(-1) caused complete growth inhibition of the four strains. Gallic acid was effective at 200, and 400 MUg mL(-1) against S. aureus, and S. pyogenes, respectively, but not against the gram negative bacteria. An OMW fraction called AntiSolvent was obtained after the addition of ethanol to the crude OMW. HPLC analysis of AntiSolvent fraction revealed that this fraction contains mainly hydroxytyrosol (10.3%), verbascoside (7.4%), and tyrosol (2.6%). The combinations of AntiSolvent/gallic acid were tested using the low minimal inhibitory concentrations which revealed that 50/100-100/100 MUg mL(-1) caused complete growth inhibition of the four strains. These results suggest that OMW specific fractions augmented with natural phenolic ingredients may be utilized as a source of bioactive compounds to control pathogenic bacteria. PMID- 21647316 TI - Prosthetic aortic valve stenosis in end-stage renal failure. AB - Although renal failure is one of the known comorbidities associated with rapid progression of aortic stenosis, it is unclear whether hemodialysis alters the progression of prosthetic aortic valve stenosis. We describe a 79-year-old female who underwent bioprosthetic aortic valve replacement 8 years ago with stable prosthetic valve area for the initial 6 years. In the last two years, coinciding with the initiation of maintenance hemodialysis, she developed progressive prosthetic valve stenosis to the point of clinical decompensation. She underwent a second prosthetic aortic valve replacement with symptom resolution. This case suggests that circulating milieu in end-stage renal failure and dialysis can accelerate the progression of prosthetic aortic valve stenosis. More frequent clinical followup and surveillance echocardiogram for dialysis patients with bioprosthetic aortic valve may facilitate timely management of valvular stenosis. PMID- 21647317 TI - Thujone-Rich Fraction of Thuja occidentalis Demonstrates Major Anti-Cancer Potentials: Evidences from In Vitro Studies on A375 Cells. AB - CRUDE ETHANOLIC EXTRACT OF THUJA OCCIDENTALIS (FAM: Cupressaceae) is used as homeopathic mother tincture (TOPhi) to treat various ailments, particularly moles and tumors, and also used in various other systems of traditional medicine. Anti proliferative and apoptosis-inducing properties of TOPhi and the thujone-rich fraction (TRF) separated from it have been evaluated for their possible anti cancer potentials in the malignant melanoma cell line A375. On initial trial by S diphenyltetrazolium bromide assay, both TOPhi and TRF showed maximum cytotoxic effect on A375 cell line while the other three principal fractions separated by chromatography had negligible or no such effect, because of which only TRF was further characterized and subjected to certain other assays for determining its precise anti-proliferative and apoptotic potentials. TRF was reported to have a molecular formula of C(10)H(16)O with a molecular weight of 152. Exposure of TRF of Thuja occidentalis to A375 cells in vitro showed more cytotoxic, anti proliferative and apoptotic effects as compared with TOPhi, but had minimal growth inhibitory responses when exposed to normal cells (peripheral blood mononuclear cell). Furthermore, both TOPhi and TRF also caused a significant decrease in cell viability, induced inter-nucleosomal DNA fragmentation, mitochondrial transmembrane potential collapse, increase in ROS generation, and release of cytochrome c and caspase-3 activation, all of which are closely related to the induction of apoptosis in A375 cells. Thus, TRF showed and matched all the anti-cancer responses of TOPhi and could be the main bio-active fraction. The use of TOPhi in traditional medicines against tumors has, therefore, a scientific basis. PMID- 21647318 TI - Acute childhood cardiorenal syndrome and impact of cardiovascular morbidity on survival. AB - Cardiorenal syndrome (CRS) clinical types, prevalence, aetiology, and acute cardiovascular morbidity impact on the outcome of acute kidney function perturbation were determined. Forty-seven of 101 (46.53%) patients with perturbed kidney function had CRS. Types 3 and 5 CRS were found in 10 and 37 patients, respectively. Type 3 CRS was due to acute glomerulonephritis (AGN; n = 7), captopril (n = 1), frusemide (n = 1), and hypovolaemia (n = 1). Malaria associated haemoglobinuria (n = 20), septicaemia (n = 11), lupus nephritis (n = 3), tumour lysis syndrome (n = 2), and acute lymphoblastic leukaemia (n = 1) caused Type 5 CRS. The cumulative mortality in hypertensive CRS was similar to nonhypertensive CRS (51.4% versus 40.9%; P = .119). Mortality in CRS and non-CRS was similar (45.7% versus 24.5%; P = .053). Type 5 survived better than type 3 CRS (66.7% versus 12.5%; P = .001). Risk factors for mortality were Type 3 CRS (P = .001), AGN-associated CRS (P = .023), dialysis requiring CRS (P = .008), and heart failure due to causes other than anaemia (P = .003). All-cause-mortality was 34.2%. Preventive measures aimed at the preventable CRS aetiologies might be critical to reducing its prevalence. PMID- 21647319 TI - Vitamin d receptor activators and clinical outcomes in chronic kidney disease. AB - Vitamin D deficiency appears to be an underestimated risk factor for cardiovascular disease in patients with chronic kidney disease. Evidence from both basic science and clinical studies supports the possible protective role of vitamin D beyond its effect on mineral metabolism. Toxicity of pharmacologic doses of active vitamin D metabolites, in particular calcitriol, is mainly due to the possibility of positive calcium and phosphorus balance. Therefore, vitamin D analogs have been developed, which suppress PTH secretion and synthesis with reduced calcemic and phosphatemic effects. Observational studies suggest that in hemodialysis patients the use of a vitamin D receptor (VDR) activator, such as calcitriol, doxercalciferol, paricalcitol, or alfacalcidol, is associated with a reduced mortality when compared with nonusers of any VDR activator. In this article the existing literature on the topic is reviewed, although a more robust answer to the question of whether or not VDR activators have beneficial effects in hemodialysis patients will hopefully come from a randomized controlled trial. PMID- 21647320 TI - Gold-catalyzed naphthalene functionalization. AB - The complexes IPrMCl (IPr = 1,3-bis(diisopropylphenyl)imidazol-2-ylidene, M = Cu, 1a; M = Au, 1b), in the presence of one equiv of NaBAr'(4) (Ar' = 3,5 bis(trifluoromethyl)phenyl), catalyze the transfer of carbene groups: C(R)CO(2)Et (R = H, Me) from N(2)C(R)CO(2)Et to afford products that depend on the nature of the metal center. The copper-based catalyst yields exclusively a cycloheptatriene derivative from the Buchner reaction, whereas the gold analog affords a mixture of products derived either from the formal insertion of the carbene unit into the aromatic C-H bond or from its addition to a double bond. In addition, no byproducts derived from carbene coupling were observed. PMID- 21647321 TI - Synthesis of chiral mono(N-heterocyclic carbene) palladium and gold complexes with a 1,1'-biphenyl scaffold and their applications in catalysis. AB - Axially chiral mono(NHC)-Pd(II) and mono(NHC)-Au(I) complexes with one side shaped 1,1'-biphenyl backbone have been prepared from chiral 6,6' dimethoxybiphenyl-2,2'-diamine. The complexes were characterized by X-ray crystal structure diffraction. The Pd(II) complex showed good catalytic activities in the Suzuki-Miyaura and Heck-Mizoroki coupling reactions, and the (S)-Au(I) complexes also showed good catalytic activities in the asymmetric intramolecular hydroamination reaction to give the corresponding product in moderate ee. PMID- 21647322 TI - A gold-catalyzed alkyne-diol cycloisomerization for the synthesis of oxygenated 5,5-spiroketals. AB - A highly efficient synthesis of oxygenated 5,5-spiroketals was performed towards the synthesis of the cephalosporolides. Gold(I) chloride in methanol induced the cycloisomerization of a protected alkyne triol with concomitant deprotection to give a strategically hydroxylated 5,5-spiroketal, despite the potential for regiochemical complications and elimination to furan. Other late transition metal Lewis acids were less effective. The use of methanol as solvent helped suppress the formation of the undesired furan by-product. This study provides yet another example of the advantages of gold catalysis in the activation of alkyne pi systems. PMID- 21647323 TI - Gold-catalyzed heterocyclizations in alkynyl- and allenyl-beta-lactams. AB - New gold-catalyzed methods using the beta-lactam scaffold have been recently developed for the synthesis of different sized heterocycles. This overview focuses on heterocyclization reactions of allenic and alkynic beta-lactams which rely on the activation of the allene and alkyne component. The mechanism as well as the regio- and stereoselectivity of the cyclizations are also discussed. PMID- 21647324 TI - Unusual behavior in the reactivity of 5-substituted-1H-tetrazoles in a resistively heated microreactor. AB - The decomposition of 5-benzhydryl-1H-tetrazole in an N-methyl-2 pyrrolidone/acetic acid/water mixture was investigated under a variety of high temperature reaction conditions. Employing a sealed Pyrex glass vial and batch microwave conditions at 240 degrees C, the tetrazole is comparatively stable and complete decomposition to diphenylmethane requires more than 8 h. Similar kinetic data were obtained in conductively heated flow devices with either stainless steel or Hastelloy coils in the same temperature region. In contrast, in a flow instrument that utilizes direct electric resistance heating of the reactor coil, tetrazole decomposition was dramatically accelerated with rate constants increased by two orders of magnitude. When 5-benzhydryl-1H-tetrazole was exposed to 220 degrees C in this type of flow reactor, decomposition to diphenylmethane was complete within 10 min. The mechanism and kinetic parameters of tetrazole decomposition under a variety of reaction conditions were investigated. A number of possible explanations for these highly unusual rate accelerations are presented. In addition, general aspects of reactor degradation, corrosion and contamination effects of importance to continuous flow chemistry are discussed. PMID- 21647325 TI - Photoinduced electron-transfer chemistry of the bielectrophoric N-phthaloyl derivatives of the amino acids tyrosine, histidine and tryptophan. AB - The photochemistry of phthalimide derivatives of the electron-rich amino acids tyrosine, histidine and tryptophan 8-10 was studied with respect to photoinduced electron-transfer (PET) induced decarboxylation and Norrish II bond cleavage. Whereas exclusive photodecarboxylation of the tyrosine substrate 8 was observed, the histidine compound 9 resulted in a mixture of histamine and preferential Norrish cleavage. The tryptophan derivative 10 is photochemically inert and shows preferential decarboxylation only when induced by intermolecular PET. PMID- 21647326 TI - The impact of a nationwide antibiotic restriction program on antibiotic usage and resistance against nosocomial pathogens in Turkey. AB - PURPOSE: Antimicrobial resistance among microorganisms is a global concern. In 2003, a nationwide antibiotic restriction program (NARP) was released in Turkey. In this study we evaluated the effect of NARP on antibiotic consumption, antimicrobial resistance, and cost. MATERIALS AND METHODS: The data obtained from all of the four university hospitals, and one referral tertiary-care educational state hospital in Ankara. Antimicrobial resistance profiles of 14,233 selected microorganisms all grown in blood cultures and antibiotic consumption from 2001 to 2005 were analyzed retrospectively. RESULTS: A negative correlation was observed between the ceftriaxone consumption and the prevalence of ceftriaxone resistant E.coli and Klebsiella spp. (rho:-0.395, p:0.332 and rho:-0.627, p:0.037, respectively). The decreased usage of carbapenems was correlated with decreased carbapenems-resistant Pseudomonas spp. and Acinetobacter spp (rho:0.155, p:0.712 and rho:0.180, p:0.668, respectively for imipenem). Methicillin resistance rates of S.aureus were decreased from 44% to 41%. After two years of NARP 5,389,155.82 USD saving occurred. CONCLUSION: NARP is effective in lowering the costs and antibiotic resistance. PMID- 21647327 TI - Foramen magnum arachnoid cyst induces compression of the spinal cord and syringomyelia: case report and literature review. AB - It is very rare that a foramen magnum arachnoid cyst induces compression of the spinal cord and syringomyelia, and currently there are few treatment experiences available. Here we reported the case of a 43-year-old male patient who admitted to the hospital due to weakness and numbness of all 4 limbs, with difficulty in urination and bowel movement. MRI revealed a foramen magnum arachnoid cyst with associated syringomyelia. Posterior fossa decompression and arachnoid cyst excision were performed. Decompression was fully undertaken during surgery; however, only the posterior wall of the arachnoid cyst was excised, because it was almost impossible to remove the whole arachnoid cyst due to toughness of the cyst and tight adhesion to the spinal cord. Three months after the surgery, MRI showed a reduction in the size of the arachnoid cyst but syrinx still remained. Despite this, the symptoms of the patient were obviously improved compared to before surgery. Thus, for the treatment of foramen magnum arachnoid cyst with compression of the spinal cord and syringomyelia, if the arachnoid cyst could not be completely excised, excision should be performed as much as possible with complete decompression of the posterior fossa, which could result in a satisfying outcome. PMID- 21647328 TI - Medical decision and patient's preference: 'much ethics' and more trust always needed. AB - There is much discussion on medical ethics literature regarding the importance of the patients' right for self-determination. We discuss some of the limitations of patient's autonomy with the aim to draw attention to the ethical complexity of medical decision making in the everyday clinical practice. PMID- 21647329 TI - Gold-catalyzed alkylation of silyl enol ethers with ortho-alkynylbenzoic acid esters. AB - Unprecedented alkylation of silyl enol ethers has been developed by the use of ortho-alkynylbenzoic acid alkyl esters as alkylating agents in the presence of a gold catalyst. The reaction probably proceeds through the gold-induced in situ construction of leaving groups and subsequent nucleophilic attack on the silyl enol ethers. The generated leaving compound abstracts a proton to regenerate the silyl enol ether structure. PMID- 21647330 TI - Expression, purification and characterization of a novel soluble human thymosin alpha1 concatemer exhibited a stronger stimulation on mice lymphocytes proliferation and higher anti-tumor activity. AB - Thymosin alpha 1 (Talpha1) has immunomodulatory and anti-tumor effects in patients and has been commercialized in worldwide. An innovative technique is therefore impending to achieve high-yield expression and purification of Talpha1 to meet the increasing requirements for clinical applications. Talpha1 can enhance T cells, dendritic cells and antibody responses, and also augment an anti tumor immune response. In the current study, we developed a novel technique to produce Talpha1 concatemer and investigated its capability in anti-tumor immunotherapy. We expressed the recombinant 2*Talpha1 concatemer protein (Talpha12 protein) in Escherichia coli. The purity of Talpha12 was higher than 95% as assessed by HPLC analysis. In vitro, Talpha12 could stimulate the proliferation of mouse splenic lymphocyte, and increase the apoptosis of tumor cell lines. In vivo, Talpha12 significantly inhibited the tumor growth in B16 tumor-bearing mice. Compared with Talpha1, the Talpha12 is of more effective bioactivity than Talpha1. The purified Talpha12 is a promising substitute for synthetic Talpha1 because of its potent anti-tumor effects. We concluded that the expression system for Talpha1 concatemer was constructed successfully, which could serves as a highly efficient tool for the production of large quantities of the highly active protein. PMID- 21647331 TI - Coupling in vitro and in vivo paradigm reveals a dose dependent inhibition of angiogenesis followed by initiation of autophagy by C6-ceramide. AB - The activity of N-hexanoyl-D-erythro-sphingosine, a C6-ceramide against angiogenesis was tested in vitro and in vivo. The effect of ceramide in inhibiting MCF-7 cancer cells was also determined. The aim of this study was to potentiate the effect of ceramide as anti-angiogenic compound that can regulate tumor induced angiogenesis.C6-ceramide inhibited vascular endothelial growth factor (VEGF)-induced human umbilical vein endothelial cells (HUVEC) tube formation in a dose-dependent manner within 24 hours. Ceramide at concentrations between 12.5 and 25 MUM inhibited the viability of MCF-7 cells and reduced VEGF induced cell migration in 24 hours. At 50 MUM, ceramide induced MCF-7 cell death via autophagy as demonstrated by accumulation of MDC in ceramide-treated MCF-7 vacuoles. The expression of VEGF was reduced and the levels of cathepsin D in MCF 7 increased. In vivo, 50 MUM ceramide caused a 40% reduction of new vessel formation in the CAM assay within 24 hours. Zebrafish exposed to 100 - 400 MUM ceramide had a distinct disruption of blood vessel development at 48 hours post fertilization. Ceramide-exposed embryos also had primary motoneurons exhibiting abnormal axonal trajectories and ectopic branching. Ceramide induced cell-death was not detected in the zebrafish assay. Collectively, these data indicate that ceramide is a potent anti-angiogenic compound and that the mechanism underlying its anti-angiogenic capabilities does not rely upon the induction of apoptosis. PMID- 21647332 TI - Mechanism of AMPK suppression of LXR-dependent Srebp-1c transcription. AB - Activation of AMP-activated protein kinase (AMPK) inhibits hepatic fatty acid synthesis by suppressing sterol regulatory element-binding protein (SREBP)-1c, a master regulator of hepatic lipogenic gene expression. Using a model cell line rat hepatoma McA-RH7777 (CRL-1601) that mimics the behavior of the intact liver by producing high levels of SREPB-1c mRNA and protein, we previously showed that AMPK suppresses hepatic Srebp-1c transcription by inhibiting endogenous liver X receptor (LXR) ligand production and SREBP-1c processing. However, whether AMPK directly inhibits ligand-induced LXR activity remained undetermined. In this study we used a series of mutant Srebp-1c promoter linked to a luciferase reporter to determine the inhibitory mechanism in rat hepatoma McA-RH7777 cells. AMPK activation by either AICAR or metformin decreases Srebp-1c promoter activity by about 75%. Normally, the synthetic LXR ligand T0901317 compound increases the wild-type Srebp-1c promoter activity by about 3-fold, which is similar to that observed in the presence of AICAR or metformin. When endogenous LXR ligand production was blocked by the potent HMG CoA reductase inhibitor compactin, T0901317-induced Srebp-1c promoter activity was decreased by AICAR or metformin treatment. In the mutant Srebp-1c promoter in which two LXR elements are intact but the sterol regulatory element (SRE) is disrupted, the fold inductions of the promoter activity by T0901317 without AMPK activators are significantly higher than those with AMPK activators. Furthermore, AMPK activation attenuates induction of endogenous SREBP-1c mRNA by T0901317. These results indicate that AMPK directly inhibits ligand-induced LXR activity in addition to blocking production of endogenous LXR ligands. PMID- 21647333 TI - Dual roles of immune cells and their factors in cancer development and progression. AB - Traditional wisdom holds that intact immune responses, such as immune surveillance or immunoediting, are required for preventing and inhibiting tumor development; but recent evidence has also indicated that unresolved immune responses, such as chronic inflammation, can promote the growth and progression of cancer. Within the immune system, cytotoxic CD8(+) and CD4(+) Th1 T cells, along with their characteristically produced cytokine IFN-gamma, function as the major anti-tumor immune effector cells, whereas tumor associated macrophages (TAM) or myeloid-derived suppressive cells (MDSC) and their derived cytokines IL 6, TNF, IL-1beta and IL-23 are generally recognized as dominant tumor-promoting forces. However, the roles played by Th17 cells, CD4(+) CD25(+) Foxp3(+) regulatory T lymphocytes and immunoregulatory cytokines such as TGF-beta in tumor development and survival remain elusive. These immune cells and the cellular factors produced from them, including both immunosuppressive and inflammatory cytokines, play dual roles in promoting or discouraging cancer development, and their ultimate role in cancer progression may rely heavily on the tumor microenvironment and the events leading to initial propagation of carcinogenesis. PMID- 21647334 TI - The heparan and heparin metabolism pathway is involved in regulation of fatty acid composition. AB - Six genes involved in the heparan sulfate and heparin metabolism pathway, DSEL (dermatan sulfate epimerase-like), EXTL1 (exostoses (multiple)-like 1), HS6ST1 (heparan sulfate 6-O-sulfotransferase 1), HS6ST3 (heparan sulfate 6-O sulfotransferase 3), NDST3 (N-deacetylase/N-sulfotransferase (heparan glucosaminyl) 3), and SULT1A1 (sulfotransferase family, cytosolic, 1A, phenol preferring, member 1), were investigated for their associations with muscle lipid composition using cattle as a model organism. Nineteen single nucleotide polymorphisms (SNPs)/multiple nucleotide length polymorphisms (MNLPs) were identified in five of these six genes. Six of these mutations were then genotyped on 246 Wagyu x Limousin F(2) animals, which were measured for 5 carcass, 6 eating quality and 8 fatty acid composition traits. Association analysis revealed that DSEL, EXTL1 and HS6ST1 significantly affected two stearoyl-CoA desaturase activity indices, the amount of conjugated linoleic acid (CLA), and the relative amount of saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA) in skeletal muscle (P<0.05). In particular, HS6ST1 joined our previously reported SCD1 and UQCRC1 genes to form a three gene network for one of the stearoyl-CoA desaturase activity indices. These results provide evidence that genes involved in heparan sulfate and heparin metabolism are also involved in regulation of lipid metabolism in bovine muscle. Whether the SNPs affected heparan sulfate proteoglycan structure is unknown and warrants further investigation. PMID- 21647335 TI - Preventing the establishment of a wildlife disease reservoir: a case study of bovine tuberculosis in wild deer in Minnesota, USA. AB - Bovine tuberculosis (bTB) has been found in 12 cattle operations and 27 free ranging white-tailed deer (Odocoileus virginianus) in northwestern Minnesota, following the state's most recent outbreak of the disease in 2005 in the northwest part of the state. Both deer and cattle have the same strain of bTB. The Minnesota Board of Animal Health has been leading efforts to eradicate the disease in Minnesota's cattle, which have included the depopulation of all infected herds, a cattle buy-out program, and mandatory fencing of stored feeds. The Minnesota Department of Natural Resources began surveillance efforts in free ranging white-tailed deer in fall 2005. All bTB-infected deer have been found within a 16 km(2) area in direct association with infected cattle farms. Aggressive efforts to reduce deer densities through liberalized hunting and sharpshooting have resulted in a 55% decline in deer densities. Also, recreational feeding of wild deer has been banned. Disease prevalence in deer has decreased from 1.2% in 2005 to an undetectable level in 2010. PMID- 21647336 TI - The First Report of Mycobacterium celatum Isolation from Domestic Pig (Sus scrofa domestica) and Roe Deer (Capreolus capreolus) and an Overview of Human Infections in Slovenia. AB - Mycobacterium celatum, a slowly growing potentially pathogenic mycobacterium first described in humans, is regarded as an uncommon cause of human infection, though capable of inducing invasive disease in immunocompromised hosts. According to some reports, a serious disease due to M. celatum may also occur in individuals with no apparent immunodeficiency. In animals, an M. celatum-related disease has been described in three cases only: twice in a domestic ferret (Mustela putorius furo) and once in a white-tailed trogon (Trogon viridis). In this paper, we report the first detection of M. celatum in a domestic pig (Sus scrofa domestica) and roe deer (Capreolus capreolus). A nation-wide overview of human M. celatum infections recorded in Slovenia between 2000 and 2010 is also given. Pulmonary disease due to M. celatum was recognized in one patient with a history of a preexisting lung disease. PMID- 21647337 TI - Ultrasonographic imaging of normal and impacted omasum in Indian crossbred cows. AB - Omasal impaction is a serious disease problem in cattle in India, but it is difficult to diagnose clinically. Ultrasonography has been proposed for the noninvasive evaluation of omasal disease. The objectives of this study were to compare the in vitro and in vivo ultrasonographic appearance of the omasum and to compare omasal appearance, limits, and size in clinically healthy cows with those in cows having confirmed omasal impaction. A 3.5 MHz curvilinear transducer was used to image and record the appearance of the omasum in vitro in a water bath, and its appearance, dorsal and ventral limits, and size in 10 healthy Indian Jersey/Red Sindhi crossbred cows. The results were compared with the ultrasonographic data collected from 5 cows with omasal impaction, as confirmed at necropsy. On moving the transducer dorsoventrally in each intercostal space and below the costal arch, the wall of omasum could be seen as an echogenic arc like structure. The difference between mean dorsoventral extents of the normal and impacted omasums was statistically insignificant. These results suggest that ultrasonographic imaging may not be useful in the diagnosis of omasal impaction in Indian crossbred cows, however, additional studies may be warranted. PMID- 21647338 TI - An investigation comparing the efficacy of topical ocular application of tacrolimus and cyclosporine in dogs. AB - The purpose of this paper was to determine the efficacy and safety of topical tacrolimus, compared to cyclosporine, for treating keratoconjunctivitis sicca (KCS) in dogs. This study was a two-phase, randomized, controlled, masked clinical trial. Phase 1 evaluated ophthalmic 0.03% tacrolimus in normal dogs. Ocular examinations were performed daily. Phase 2 evaluated the efficacy of tacrolimus in treating KCS. Half the dogs received 2% cyclosporine A; the others received 0.03% tacrolimus, both diluted in olive oil. Four ophthalmic examinations were done over 12 weeks. There was no significant difference between groups in phase I. In phase 2, there was no significant difference in Schirmer tear test I (STT) results between the two groups, and both groups had a significant increase in STT over time. Both drugs were effective in increasing the STT in dogs naive to lacrimostimulants. Tacrolimus was effective in increasing the STT in 4 dogs currently nonresponsive to cyclosporine. PMID- 21647339 TI - Clinical Evaluation of Xylazine-Butorphanol-Guaifenesin-Ketamine as Short-Term TIVA in Equines. AB - Xylazine (1.1 mg/kg), butorphanol (0.02 mg/kg), guaifenesin 5% (20 mg/kg), and ketamine (2.2 mg/kg) combinations were able to induce short-term surgical anaesthesia for 23.33 +/- 2.57 min in Spiti ponies with excellent to good muscle relaxation and analgesia. Urination and neighing was a constant feature during recovery period. Biphasic P wave, biphasic T wave, and depressed PR segment were common electrocardiographic findings. Biochemical attributes were within physiological limits except a significant increase in ALT values during anaesthesia. The values returned to normal during recovery ruling out any renal or hepatic toxicity. Occasionally, negative T wave, notched P wave, and sinus block were noticed. It is recommended that the anaesthetic combinations xylazine butorphanol-guaifenesin-ketamine can be safely used for short-term total intravenous anaesthesia (TIVA) in equines under field conditions. PMID- 21647340 TI - Ram Sperm Motility Parameters under The Influence of Epidermal Growth Factor. AB - Epidermal growth factor (EGF) is one of the important cytokines that play a role in fertility. It is known that EGF affects both male and female reproduction, but its effect on sperm parameters is not fully understood. Up to the present, the effect of EGF on ram sperm motility parameters has not been published. We analyzed motility parameters of ejaculates after 24, 48, and 72 hours from the EGF addition. EGF was added to chilled ram sperm at concentrations of 0, 100, 200, and 400 ng.ml(-1). Analyses were realized using computer, assisted semen analyzer (CASA)-Hamilton Thorn motility analyzer (version 7). The effect of EGF was already visible after 30 min of incubation. Significant effect on ram sperm total motility and progressive movement was observed at higher EGF concentrations after 48 h of incubation. Our results show that EGF affects sperm motility parameters depending on concentration and time of exposure. PMID- 21647341 TI - Bovine tuberculosis and the establishment of an eradication program in the United States: role of veterinarians. AB - The significance of the identification of Mycobacterium bovis as a zoonotic pathogen in 1882 was not initially recognized. After years of research by veterinarians, and other scientists, the importance of M. bovis as a pathogen and the public health ramifications, were appreciated. Veterinarians played pivotal roles in the creation of improved meat and milk inspection, diagnosis of M. bovis infected cattle, and in time, a bovine tuberculosis eradication program that would impact every cattle producer in the country. After overcoming many challenges, the 93-year-long program has decreased disease prevalence from 5% to <0.001%. Today, years of hard work by practitioners, researchers and regulatory officials alike, have yielded a program with a net benefit of almost $160 million per year. PMID- 21647342 TI - Ovarian Follicular Dynamics during the Estrous Cycle in Jennies in Upper Egypt. AB - The objective of the current study was to describe follicular dynamics in Egyptian Jennies throughout the estrous cycle. In this experiment, 8 estrus cycles in 8 cyclic Jennies were studied from February to June using ultrasonography. The result revealed that one follicular wave per cycle was recorded throughout the studied period. Dominant follicle (DF) was firstly detected at -0.80 +/- 0.84 day in Jennies. The growth rate of DF was 2.32 +/- 0.18 mm/day. Left ovulations were nonsignificantly (P = .07) more than right ovulations (55.6% versus 44.6%). The CL was firstly detected at D 2.58 +/- 1.2, developed in a rate of 1.19 +/- 0.07 mm/day, reached a maximum diameter of 30.77 +/- 1.28 mm at D 13.0 +/- 0.70, and started to regress on D 17.05 +/- 0.64 with a mean regression rate of 1.75 +/- 0.17 mm d(-1). Results of the present study indicated that Jennies had one follicular wave per cycle. The Day of the cycle has a significant effect on the number of different classes of the ovarian follicles, but not large ones. Ultrasonographic characteristics of the preovulatory follicles could be useful to predict ovulation. CL developed and regressed in a slow rate. PMID- 21647343 TI - Overt Mycobacterium avium subsp. paratuberculosis Infection: An Infrequent Occurrence in Archived Tissue from False TB Reactor Cattle in Michigan, USA. AB - The objective of this study was to retrospectively determine whether or not cattle from the state of Michigan which were classified as bovine tuberculosis reactors, based on currently approved field and laboratory testing methods, were overtly infected with Mycobacterium avium subsp. paratuberculosis (MAP). Included in this study were 384 adult cattle submitted to the Diagnostic Center for Population and Animal Health over a seven-year period. Cattle were tested utilizing standard methods to confirm that all cattle were lesion and culture negative for infection with Mycobacterium bovis at postmortem examination. Retrospective analysis of formalin-fixed, paraffin-embedded sections of ileum and ileocecal lymph node were evaluated by histopathology, acid-fast staining, and PCR assays to detect MAP. Overall, only 1.04 percent of cattle showed overt infection with MAP on visual examination of sections of ileum and/or ileo-cecal lymph node. This increased slightly to 2.1 percent of cattle likely infected with MAP after additional testing using a PCR assay. Based on these results, we found no evidence that overt infection with MAP plays a major role in the false tuberculosis reactor test results for cattle examined in this study. PMID- 21647344 TI - Isolation and genetic analysis of bovine viral diarrhea virus from infected cattle in indiana. AB - Species and biotype distribution was determined in 44 bovine viral diarrhea virus (BVDV-) positive samples submitted to the Animal Disease Diagnostic Laboratory (ADDL) in Indiana during 2006-2008. BVDV RNA was detected in the 5'-untranslated region and N(pro) region using reverse transcriptase PCR followed by sequencing analysis of the PCR product. Additionally, cases were classified into one of six categories according to history and/or lesions: acute symptomatic, hemorrhagic, respiratory distress, reproductive, persistent infection (PI), and mucosal disease (MD). Of 44 BVDV-positive samples, 33 were noncytopathic (ncp), 10 were cytopathic (cp), and one presented both ncp and cp biotypes. Sequencing analysis demonstrated that all samples belonged to BVDV-1a, BVDV-1b, or BVDV-2. The most common isolate was ncp BVDV-1b, (44%) followed by ncp BVDV-2a (24%). Among the six categories, respiratory clinical signs were the most common (36%) followed by PI (25%) and MD (16%). PMID- 21647345 TI - A Dry EEG-System for Scientific Research and Brain-Computer Interfaces. AB - Although it ranks among the oldest tools in neuroscientific research, electroencephalography (EEG) still forms the method of choice in a wide variety of clinical and research applications. In the context of brain-computer interfacing (BCI), EEG recently has become a tool to enhance human-machine interaction. EEG could be employed in a wider range of environments, especially for the use of BCI systems in a clinical context or at the homes of patients. However, the application of EEG in these contexts is impeded by the cumbersome preparation of the electrodes with conductive gel that is necessary to lower the impedance between electrodes and scalp. Dry electrodes could provide a solution to this barrier and allow for EEG applications outside the laboratory. In addition, dry electrodes may reduce the time needed for neurological exams in clinical practice. This study evaluates a prototype of a three-channel dry electrode EEG system, comparing it to state-of-the-art conventional EEG electrodes. Two experimental paradigms were used: first, event-related potentials (ERP) were investigated with a variant of the oddball paradigm. Second, features of the frequency domain were compared by a paradigm inducing occipital alpha. Furthermore, both paradigms were used to evaluate BCI classification accuracies of both EEG systems. Amplitude and temporal structure of ERPs as well as features in the frequency domain did not differ significantly between the EEG systems. BCI classification accuracies were equally high in both systems when the frequency domain was considered. With respect to the oddball classification accuracy, there were slight differences between the wet and dry electrode systems. We conclude that the tested dry electrodes were capable to detect EEG signals with good quality and that these signals can be used for research or BCI applications. Easy to handle electrodes may help to foster the use of EEG among a wider range of potential users. PMID- 21647346 TI - Administrative censoring in ecological analyses of autism and a Bayesian solution. AB - Widely cited ecological analyses of autism have reported associations with mercury emissions, with precipitation, and race at the level of counties or school districts. However, state educational agencies often suppress any low numerical autism counts before releasing data--a phenomenon known as "administrative censoring." Previous analyses did not describe appropriate methods for censored data analysis; common substitution or exclusion methods are known to introduce bias and produce artificially narrow confidence intervals. We apply a Bayesian censored random effects Poisson model to reanalyze associations between 2001 Toxic Release Inventory reported mercury emissions and 2000-2001 autism counts in Texas. Relative risk estimates for autism decreased from 4.44 (95% CI: 4.16, 4.74) per thousand lbs. of air mercury emissions using a naive zero-substitution approach to 1.42 (95% CI: 1.09, 1.78) using the Bayesian approach. Inadequate attention to censoring poses a serious threat to the validity of ecological analyses of autism and other health outcomes. PMID- 21647347 TI - Liver transplantation in an adult with citrullinaemia type 2. AB - Citrullinaemia is a urea cycle defect that results from a deficiency of the enzyme arginosuccinate synthetase. Type 1 disease is diagnosed in childhood, whereas Type 2 disease is adult onset. We report the outcome of a patient with citrullinemia Type 2 who received a liver transplant at our center and the implications of this diagnosis in liver transplantation. PMID- 21647348 TI - Hemotoxicity induced by chronic chlorpyrifos exposure in wistar rats: mitigating effect of vitamin C. AB - The study evaluated the ameliorative effect of vitamin C on chronic chlorpyrifos induced hematological alterations in Wistar rats. Twenty adult male rats divided into 4 groups of 5 animals each were exposed to the following regimens: group I (S/oil) was administered soya oil (2 mL/kg b.w.), while group II (VC) was given vitamin C (100 mg/kg b.w.); group III was dosed with CPF (10.6 mg/kg b.w.); group IV was pretreated with vitamin C (100 mg/kg) and then exposed to CPF (10.6 mg/kg b.w.), 30 minutes later. The regimens were administered by oral gavage once daily for a period of 17 weeks. Blood samples collected at the end of the study revealed reduction in the levels of pack cell volume, hemoglobin, red blood cells, leukocytes (attributed to neutropenia, lymphopenia, and monocytopenia), and platelets in the CPF group, which were ameliorated in the vitamin C- pretreated group. The elevated values of malonaldehyde, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, and neutrophil/lymphocyte ratio in the CPF group were restored in those pretreated with vitamin C. The study has shown that chronic CPF-induced adversity on hematological parameters of Wistar rats was mitigated by pretreatment with vitamin C. PMID- 21647349 TI - Impact of acute rejection on kidney allograft outcomes in recipients on rapid steroid withdrawal. AB - Background. Our aim was to study the impact of clinical acute rejection (CR) and subclinical rejection (SR) on outcomes in kidney transplant recipients treated with rapid steroid withdrawal (RSW). Methods. All patients who received a living or deceased donor kidney transplant and were treated with RSW were included. The primary outcome was death-censored graft survival. Biopsies with Banff borderline changes were included with the rejection groups. Results. 457 kidney transplant recipients treated with RSW were included; 46 (10%) experienced SR, and 36 (7.8%) had CR. Mean HLA mismatch was significantly higher in the CR group. The Banff grade of rejection was higher in the CR group. There was a larger proportion of patients in both rejection groups with the combination of IFTA and persistent inflammation on the follow-up protocol biopsy done at 1 year. The estimated 5 year death-censored graft survival was 81% in SR, 78% in CR, and 97% in the control group (P < .0001). Significant differences were observed in allograft survival between the CR and control group (HR 9.06, 95% CI 3.39-24.2) and between the SR and control group (HR 4.22, 95% CI 1.30-13.7). Conclusion. Both SR and CR are associated with an inferior graft survival in recipients on RSW. PMID- 21647350 TI - Persistent organic pollutants in serum and several different fat compartments in humans. AB - BACKGROUND: Chemicals that store in lipid-rich compartments have the potential for long-term disruption of metabolic and endocrine processes. Given the evidence that persistent organic pollutants (POPs) also alter systemic metabolic, endocrine, and immune system functions, it follows that elevated chemical concentrations in intra-abdominal fat may alter function, through local chemical signaling, of visceral organs. Despite this potential, there has been little study defining POP concentrations in live human intra-abdominal fat. It is at present uncertain whether POPs distribute equally to all fat compartments, including fat in serum. METHODS: Seven human subjects scheduled for elective surgery for benign lesions or cancer provided consent for removal of samples of subcutaneous and intra-abdominal fat and/or cancerous tissue. These samples were analyzed for 22 chlorinated pesticides and 10 polychlorinated biphenyl (PCB) congeners by GC/ECD plus GC/MS. RESULTS: In only two subjects were the patterns and relative concentrations of PCBs and pesticides about the same in all fat compartments. In the other subjects, there were major differences in levels in subcutaneous as compared to other compartments, but with some higher and some lower. While the pattern of PCBs in the various compartments matched that of the pesticides in some, it was opposite in others. INTERPRETATION: These results demonstrate a complicated distribution of PCB congeners and pesticides in various lipid compartments. The difference may reflect various K(ow)s, different rates of metabolism, and/or different lengths of exposure. But the results suggest that contaminant levels in serum or even subcutaneous fat do not necessarily indicate concentrations and patterns in other kinds of adipose tissue. PMID- 21647351 TI - The use of mixed models for the analysis of mediated data with time-dependent predictors. AB - Linear mixed models (LMMs) are frequently used to analyze longitudinal data. Although these models can be used to evaluate mediation, they do not directly model causal pathways. Structural equation models (SEMs) are an alternative technique that allows explicit modeling of mediation. The goal of this paper is to evaluate the performance of LMMs relative to SEMs in the analysis of mediated longitudinal data with time-dependent predictors and mediators. We simulated mediated longitudinal data from an SEM and specified delayed effects of the predictor. A variety of model specifications were assessed, and the LMMs and SEMs were evaluated with respect to bias, coverage probability, power, and Type I error. Models evaluated in the simulation were also applied to data from an observational cohort of HIV-infected individuals. We found that when carefully constructed, the LMM adequately models mediated exposure effects that change over time in the presence of mediation, even when the data arise from an SEM. PMID- 21647352 TI - Inward leakage in tight-fitting PAPRs. AB - A combination of local flow measurement techniques and fog flow visualization was used to determine the inward leakage for two tight-fitting powered air-purifying respirators (PAPRs), the 3M Breathe-Easy PAPR and the SE 400 breathing demand PAPR. The PAPRs were mounted on a breathing machine head form, and flows were measured from the blower and into the breathing machine. Both respirators leaked a little at the beginning of inhalation, probably through their exhalation valves. In both cases, the leakage was not enough for fog to appear at the mouth of the head form. PMID- 21647353 TI - Dynamics of networks of excitatory and inhibitory neurons in response to time dependent inputs. AB - We investigate the dynamics of recurrent networks of excitatory (E) and inhibitory (I) neurons in the presence of time-dependent inputs. The dynamics is characterized by the network dynamical transfer function, i.e., how the population firing rate is modulated by sinusoidal inputs at arbitrary frequencies. Two types of networks are studied and compared: (i) a Wilson-Cowan type firing rate model; and (ii) a fully connected network of leaky integrate-and fire (LIF) neurons, in a strong noise regime. We first characterize the region of stability of the "asynchronous state" (a state in which population activity is constant in time when external inputs are constant) in the space of parameters characterizing the connectivity of the network. We then systematically characterize the qualitative behaviors of the dynamical transfer function, as a function of the connectivity. We find that the transfer function can be either low-pass, or with a single or double resonance, depending on the connection strengths and synaptic time constants. Resonances appear when the system is close to Hopf bifurcations, that can be induced by two separate mechanisms: the I-I connectivity and the E-I connectivity. Double resonances can appear when excitatory delays are larger than inhibitory delays, due to the fact that two distinct instabilities exist with a finite gap between the corresponding frequencies. In networks of LIF neurons, changes in external inputs and external noise are shown to be able to change qualitatively the network transfer function. Firing rate models are shown to exhibit the same diversity of transfer functions as the LIF network, provided delays are present. They can also exhibit input dependent changes of the transfer function, provided a suitable static non linearity is incorporated. PMID- 21647354 TI - Alcohol marketing, drunkenness, and problem drinking among Zambian youth: findings from the 2004 Global School-Based Student Health Survey. AB - This study examines the associations between alcohol marketing strategies, alcohol education including knowledge about dangers of alcohol and refusal of alcohol, and drinking prevalence, problem drinking, and drunkenness. Analyses are based on the Global School-Based Student Health Survey (GSHS) conducted in Zambia (2004) of students primarily 11 to 16 years of age (N = 2257). Four statistical models were computed to test the associations between alcohol marketing and education and alcohol use, while controlling for possible confounding factors. Alcohol marketing, specifically through providing free alcohol through a company representative, was associated with drunkenness (AOR = 1.49; 95% CI: 1.09-2.02) and problem drinking (AOR = 1.41; 95% CI: 1.06-1.87) among youth after controlling for demographic characteristics, risky behaviors, and alcohol education. However, alcohol education was not associated with drunkenness or problem drinking. These findings underscore the importance of restricting alcohol marketing practices as an important policy strategy for reducing alcohol use and its dire consequences among vulnerable youth. PMID- 21647355 TI - Modeling and syndromic surveillance for estimating weather-induced heat-related illness. AB - This paper compares syndromic surveillance and predictive weather-based models for estimating emergency department (ED) visits for Heat-Related Illness (HRI). A retrospective time-series analysis of weather station observations and ICD-coded HRI ED visits to ten hospitals in south eastern Ontario, Canada, was performed from April 2003 to December 2008 using hospital data from the National Ambulatory Care Reporting System (NACRS) database, ED patient chief complaint data collected by a syndromic surveillance system, and weather data from Environment Canada. Poisson regression and Fast Orthogonal Search (FOS), a nonlinear time series modeling technique, were used to construct models for the expected number of HRI ED visits using weather predictor variables (temperature, humidity, and wind speed). Estimates of HRI visits from regression models using both weather variables and visit counts captured by syndromic surveillance as predictors were slightly more highly correlated with NACRS HRI ED visits than either regression models using only weather predictors or syndromic surveillance counts. PMID- 21647356 TI - Worksite health promotion in six varied US sites: beta testing as a needed translational step. AB - BACKGROUND: Dissemination of health promotion interventions generally has followed an efficacy, effectiveness to full scale paradigm, and most programs have failed to traverse that sequence. OBJECTIVE: Report national dissemination of a health promotion program and juxtapose sequential case study observations with the current technology transfer literature. DESIGN: Multiple department level case studies using contact logs, transcribed interactions, augmented with field notes and validated by respondent review; at least two investigators independently generated site summaries, which were compared to formulate a final report. RESULTS: Adoption was facilitated with national partners and designing branded materials. Critical site influences included departmental features, local champions, and liaison relationships. Achieving distal reach and fidelity required sequential process and program revisions based on new findings at each site. CONCLUSIONS: Beta testing to redesign program elements and modify process steps appears to be a needed and often ignored translational step between efficacy and more widespread dissemination. PMID- 21647357 TI - Scalp Angiosarcoma Remission with Bevacizumab and Radiotherapy without Surgery: A Case Report and Review of the Literature. AB - Angiosarcoma (AS) is a rare and aggressive vascular neoplasm with very poor prognosis. Patients with extensive cutaneous AS who are not surgical candidates have very limited options since there is no standard treatment. Treatment options include radiation, chemotherapy, and angiogenesis inhibitor with varying success rates. Here, we report a case an 88 year old patient with extensive scalp angiosarcoma having biopsy proven remission with bevacizumab and radiotherapy without undergoing surgery. PMID- 21647358 TI - Tyrosine Phosphorylation in the C-Terminal Nuclear Localization and Retention Signal (C-NLS) of the EWS Protein. AB - Ewing sarcoma (EWS) proto-oncoprotein, an RNA-binding protein, is involved in DNA recombination and repair, gene expression, RNA processing and transport, as well as cell signalling. Chimeric EWS oncoproteins generated by chromosomal translocations between EWSR1 and the genes of transcription factors cause malignant tumors. To understand the loss of function by these translocations, the role of the intact EWS protein has to be investigated. The predominantly nuclear localization of the EWS protein via a transportin-1-mediated mechanism is dependent on the recently identified C-NLS (also known as PY-NLS). Among other residues in the C-NLS, Y656 interacts with transportin-1 and is essential for its nuclear localization. Here, we show that Y656 is phosphorylated, which seems to be a critical factor for transportin-1-mediated nuclear import. If Y656 was mutated cytosolic aggregates of the EWS protein, colocalized with transportin-1, were observed, similar to those described with mutants of the closely related FUS/TLS protein that had amino acid substitutions in the PY-NLS causing familial amyothrophic lateral sclerosis. PMID- 21647360 TI - Chondrosarcoma of the thorax. AB - Although a rare entity, chondrosarcoma is the most common malignant tumor of the chest wall. Most patients present with an enlarging, painful anterior chest wall mass arising from the costochondrosternal junction. CT scan with intravenous contrast is the gold standard radiographic study for diagnosis and operative planning. Contrary to previous dictum, resection may be performed in an appropriate surgical candidate based on imaging characteristics or image-guided percutaneous biopsy results; incisional biopsy is rarely required. The keys to successful treatment are early recognition and radical excision with adequate margins, as chondrosarcoma is relatively resistant to radiotherapy and conventional cytotoxic chemotherapy. Overall survival is excellent in most surgical series from experienced centers. Complete excision with widely negative microscopic margins at the initial operation is of the utmost importance, as local recurrence portends systemic metastasis and eventual tumor-related mortality. This paper summarizes data from relevant surgical series and thereupon draws conclusions regarding preoperative, intraoperative, and postoperative management of thoracic chondrosarcoma. PMID- 21647359 TI - Noradrenaline and Parkinson's disease. AB - Parkinson's disease (PD) is characterized by the degeneration of dopamine (DA) neurons in the substantia nigra pars compacta, and motor symptoms including bradykinesia, rigidity, and tremor at rest. These symptoms are exhibited when striatal dopamine concentration has decreased by around 70%. In addition to motor deficits, PD is also characterized by the non-motor symptoms. However, depletion of DA alone in animal models has failed to simultaneously elicit both the motor and non-motor deficits of PD, possibly because the disease is a multi-system disorder that features a profound loss in other neurotransmitter systems. There is growing evidence that additional loss of noradrenaline (NA) neurons of the locus coeruleus, the principal source of NA in the brain, could be involved in the clinical expression of motor as well as in non-motor deficits. In the present review, we analyze the latest evidence for the implication of NA in the pathophysiology of PD obtained from animal models of parkinsonism and from parkinsonian patients. Recent studies have shown that NA depletion alone, or combined with DA depletion, results in motor as well as in non-motor dysfunctions. In addition, by using selective agonists and antagonists of noradrenaline alpha receptors we, and others, have shown that alpha2 receptors are implicated in the control of motor activity and that alpha2 receptor antagonists can improve PD motor symptoms as well as l-Dopa-induced dyskinesia. In this review we argue that the loss of NA neurons in PD has an impact on all PD symptoms and that the addition of NAergic agents to dopaminergic medication could be beneficial in the treatment of the disease. PMID- 21647361 TI - Targeting the Insulin-Like Growth Factor 1 Receptor in Ewing's Sarcoma: Reality and Expectations. AB - Ewing's sarcoma family of tumours comprises a group of very aggressive diseases that are potentially curable with multimodality treatment. Despite the undoubted success of current treatment, approximately 30% of patients will relapse and ultimately die of disease. The insulin-like growth factor 1 receptor (IGF-1R) has been implicated in the genesis, growth, proliferation, and the development of metastatic disease in Ewing's sarcoma. In addition, IGF1-R has been validated, both in vitro and in vivo, as a potential therapeutic target in Ewing's sarcoma. Phase I studies of IGF-1R monoclonal antibodies reported several radiological and clinical responses in Ewing's sarcoma patients, and initial reports of several Phase II studies suggest that about a fourth of the patients would benefit from IGF-1R monoclonal antibodies as single therapy, with approximately 10% of patients achieving objective responses. Furthermore, these therapies are well tolerated, and thus far severe toxicity has been rare. Other studies assessing IGF-1R monoclonal antibodies in combination with traditional cytotoxics or other targeted therapies are expected. Despite, the initial promising results, not all patients benefit from IGF-1R inhibition, and consequently, there is an urgent need for the identification of predictive markers of response. PMID- 21647362 TI - The prevalence of pain in patients attending sarcoma outpatient clinics. AB - The prevalence of pain in patients with sarcoma is not well documented. We investigated this in outpatients at a tertiary cancer referral centre, assessing the adequacy of pain control and for risk factors leading to higher prevalence and severity of pain. 149 patients were surveyed. Patients with pain within the previous 7 days completed pain assessment tools (BPI, S-LANSS, PMI). 53% of patients had pain within the previous 7 days, and 25% had significant pain. Of those with pain, 63% was inadequately controlled and neuropathic pain was identified in 36%. Age, gender, tumour type, and the type of cancer treatment were not significant predictors of the prevalence or severity of the pain. Based on our results, patients with sarcoma should be actively screened for pain and have regular reviews of their analgesic requirements. PMID- 21647363 TI - The Bone Niche of Chondrosarcoma: A Sanctuary for Drug Resistance, Tumour Growth and also a Source of New Therapeutic Targets. AB - Chondrosarcomas are malignant cartilage-forming tumours representing around 20% of malignant primary tumours of bone and affect mainly adults in the third to sixth decade of life. Unfortunately, the molecular pathways controlling the genesis and the growth of chondrosarcoma cells are still not fully defined. It is well admitted that the invasion of bone by tumour cells affects the balance between early bone resorption and formation and induces an "inflammatory-like" environment which establishes a dialogue between tumour cells and their environment. The bone tumour microenvironment is then described as a sanctuary that contributes to the drug resistance patterns and may control at least in part the tumour growth. The concept of "niche" defined as a specialized microenvironment that can promote the emergence of tumour stem cells and provide all the required factors for their development recently emerges in the literature. The present paper aims to summarize the main evidence sustaining the existence of a specific bone niche in the pathogenesis of chondrosarcomas. PMID- 21647364 TI - Medio-lateral postural instability in subjects with tinnitus. AB - BACKGROUND: Many patients show modulation of tinnitus by gaze, jaw or neck movements, reflecting abnormal sensorimotor integration, and interaction between various inputs. Postural control is based on multi-sensory integration (visual, vestibular, somatosensory, and oculomotor) and indeed there is now evidence that posture can also be influenced by sound. Perhaps tinnitus influences posture similarly to external sound. This study examines the quality of postural performance in quiet stance in patients with modulated tinnitus. METHODS: Twenty three patients with highly modulated tinnitus were selected in the ENT service. Twelve reported exclusively or predominately left tinnitus, eight right, and three bilateral. Eighteen control subjects were also tested. Subjects were asked to fixate a target at 40 cm for 51 s; posturography was performed with the platform (Technoconcept, 40 Hz) for both the eyes open and eyes closed conditions. RESULTS: For both conditions, tinnitus subjects showed abnormally high lateral body sway (SDx). This was corroborated by fast Fourrier Transformation (FFTx) and wavelet analysis. For patients with left tinnitus only, medio-lateral sway increased significantly when looking away from the center. CONCLUSION: Similarly to external sound stimulation, tinnitus could influence lateral sway by activating attention shift, and perhaps vestibular responses. Poor integration of sensorimotor signals is another possibility. Such abnormalities would be accentuated in left tinnitus because of the importance of the right cerebral cortex in processing both auditory-tinnitus eye position and attention. PMID- 21647365 TI - Circular permutation of red fluorescent proteins. AB - Circular permutation of fluorescent proteins provides a substrate for the design of molecular sensors. Here we describe a systematic exploration of permutation sites for mCherry and mKate using a tandem fusion template approach. Circular permutants retaining more than 60% (mCherry) and 90% (mKate) brightness of the parent molecules are reported, as well as a quantitative evaluation of the fluorescence from neighboring mutations. Truncations of circular permutants indicated essential N- and C-terminal segments and substantial flexibility in the use of these molecules. Structural evaluation of two cp-mKate variants indicated no major conformational changes from the previously reported wild-type structure, and cis conformation of the chromophores. Four cp-mKates were identified with over 80% of native fluorescence, providing important new building blocks for sensor and complementation experiments. PMID- 21647366 TI - In silico and in vitro investigations of the mutability of disease-causing missense mutation sites in spermine synthase. AB - BACKGROUND: Spermine synthase (SMS) is a key enzyme controlling the concentration of spermidine and spermine in the cell. The importance of SMS is manifested by the fact that single missense mutations were found to cause Snyder-Robinson Syndrome (SRS). At the same time, currently there are no non-synonymous single nucleoside polymorphisms, nsSNPs (harmless mutations), found in SMS, which may imply that the SMS does not tolerate amino acid substitutions, i.e. is not mutable. METHODOLOGY/PRINCIPAL FINDINGS: To investigate the mutability of the SMS, we carried out in silico analysis and in vitro experiments of the effects of amino acid substitutions at the missense mutation sites (G56, V132 and I150) that have been shown to cause SRS. Our investigation showed that the mutation sites have different degree of mutability depending on their structural micro environment and involvement in the function and structural integrity of the SMS. It was found that the I150 site does not tolerate any mutation, while V132, despite its key position at the interface of SMS dimer, is quite mutable. The G56 site is in the middle of the spectra, but still quite sensitive to charge residue replacement. CONCLUSIONS/SIGNIFICANCE: The performed analysis showed that mutability depends on the detail of the structural and functional factors and cannot be predicted based on conservation of wild type properties alone. Also, harmless nsSNPs can be expected to occur even at sites at which missense mutations were found to cause diseases. PMID- 21647367 TI - Role of Kv1 potassium channels in regulating dopamine release and presynaptic D2 receptor function. AB - Dopamine (DA) release in the CNS is critical for motor control and motivated behaviors. Dysfunction of its regulation is thought to be implicated in drug abuse and in diseases such as schizophrenia and Parkinson's. Although various potassium channels located in the somatodendritic compartment of DA neurons such as G-protein-gated inward rectifying potassium channels (GIRK) have been shown to regulate cell firing and DA release, little is presently known about the role of potassium channels localized in the axon terminals of these neurons. Here we used fast-scan cyclic voltammetry to study electrically-evoked DA release in rat dorsal striatal brain slices. We find that although G-protein-gated inward rectifying (GIRK) and ATP-gated (K(ATP)) potassium channels play only a minor role, voltage-gated potassium channels of the Kv1 family play a major role in regulating DA release. The use of Kv subtype-selective blockers confirmed a role for Kv1.2, 1.3 and 1.6, but not Kv1.1, 3.1, 3.2, 3.4 and 4.2. Interestingly, Kv1 blockers also reduced the ability of quinpirole, a D2 receptor agonist, to inhibit evoked DA overflow, thus suggesting that Kv1 channels also regulate presynaptic D2 receptor function. Our work identifies Kv1 potassium channels as key regulators of DA release in the striatum. PMID- 21647368 TI - Quantitative detection and biological propagation of scrapie seeding activity in vitro facilitate use of prions as model pathogens for disinfection. AB - Prions are pathogens with an unusually high tolerance to inactivation and constitute a complex challenge to the re-processing of surgical instruments. On the other hand, however, they provide an informative paradigm which has been exploited successfully for the development of novel broad-range disinfectants simultaneously active also against bacteria, viruses and fungi. Here we report on the development of a methodological platform that further facilitates the use of scrapie prions as model pathogens for disinfection. We used specifically adapted serial protein misfolding cyclic amplification (PMCA) for the quantitative detection, on steel wires providing model carriers for decontamination, of 263K scrapie seeding activity converting normal protease-sensitive into abnormal protease-resistant prion protein. Reference steel wires carrying defined amounts of scrapie infectivity were used for assay calibration, while scrapie contaminated test steel wires were subjected to fifteen different procedures for disinfection that yielded scrapie titre reductions of <=10(1)- to >=10(5.5)-fold. As confirmed by titration in hamsters the residual scrapie infectivity on test wires could be reliably deduced for all examined disinfection procedures, from our quantitative seeding activity assay. Furthermore, we found that scrapie seeding activity present in 263K hamster brain homogenate or multiplied by PMCA of scrapie-contaminated steel wires both triggered accumulation of protease resistant prion protein and was further propagated in a novel cell assay for 263K scrapie prions, i.e., cerebral glial cell cultures from hamsters. The findings from our PMCA- and glial cell culture assays revealed scrapie seeding activity as a biochemically and biologically replicative principle in vitro, with the former being quantitatively linked to prion infectivity detected on steel wires in vivo. When combined, our in vitro assays provide an alternative to titrations of biological scrapie infectivity in animals that substantially facilitates the use of prions as potentially highly indicative test agents in the search for novel broad-range disinfectants. PMID- 21647369 TI - Reelin controls progenitor cell migration in the healthy and pathological adult mouse brain. AB - Understanding the signals that control migration of neural progenitor cells in the adult brain may provide new therapeutic opportunities. Reelin is best known for its role in regulating cell migration during brain development, but we now demonstrate a novel function for reelin in the injured adult brain. First, we show that Reelin is upregulated around lesions. Second, experimentally increasing Reelin expression levels in healthy mouse brain leads to a change in the migratory behavior of subventricular zone-derived progenitors, triggering them to leave the rostral migratory stream (RMS) to which they are normally restricted during their migration to the olfactory bulb. Third, we reveal that Reelin increases endogenous progenitor cell dispersal in periventricular structures independently of any chemoattraction but via cell detachment and chemokinetic action, and thereby potentiates spontaneous cell recruitment to demyelination lesions in the corpus callosum. Conversely, animals lacking Reelin signaling exhibit reduced endogenous progenitor recruitment at the lesion site. Altogether, these results demonstrate that beyond its known role during brain development, Reelin is a key player in post-lesional cell migration in the adult brain. Finally our findings provide proof of concept that allowing progenitors to escape from the RMS is a potential therapeutic approach to promote myelin repair. PMID- 21647370 TI - SNP-based typing: a useful tool to study Bordetella pertussis populations. AB - To monitor changes in Bordetella pertussis populations, mainly two typing methods are used; Pulsed-Field Gel Electrophoresis (PFGE) and Multiple-Locus Variable Number Tandem Repeat Analysis (MLVA). In this study, a single nucleotide polymorphism (SNP) typing method, based on 87 SNPs, was developed and compared with PFGE and MLVA. The discriminatory indices of SNP typing, PFGE and MLVA were found to be 0.85, 0.95 and 0.83, respectively. Phylogenetic analysis, using SNP typing as Gold Standard, revealed false homoplasies in the PFGE and MLVA trees. Further, in contrast to the SNP-based tree, the PFGE- and MLVA-based trees did not reveal a positive correlation between root-to-tip distance and the isolation year of strains. Thus PFGE and MLVA do not allow an estimation of the relative age of the selected strains. In conclusion, SNP typing was found to be phylogenetically more informative than PFGE and more discriminative than MLVA. Further, in contrast to PFGE, it is readily standardized allowing interlaboratory comparisons. We applied SNP typing to study strains with a novel allele for the pertussis toxin promoter, ptxP3, which have a worldwide distribution and which have replaced the resident ptxP1 strains in the last 20 years. Previously, we showed that ptxP3 strains showed increased pertussis toxin expression and that their emergence was associated with increased notification in The Netherlands. SNP typing showed that the ptxP3 strains isolated in the Americas, Asia, Australia and Europe formed a monophyletic branch which recently diverged from ptxP1 strains. Two predominant ptxP3 SNP types were identified which spread worldwide. The widespread use of SNP typing will enhance our understanding of the evolution and global epidemiology of B. pertussis. PMID- 21647371 TI - Cancer cell migration: integrated roles of matrix mechanics and transforming potential. AB - Significant progress has been achieved toward elucidating the molecular mechanisms that underlie breast cancer progression; yet, much less is known about the associated cellular biophysical traits. To this end, we use time-lapsed confocal microscopy to investigate the interplay among cell motility, three dimensional (3D) matrix stiffness, matrix architecture, and transforming potential in a mammary epithelial cell (MEC) cancer progression series. We use a well characterized breast cancer progression model where human-derived MCF10A MECs overexpress either ErbB2, 14-3-3zeta, or both ErbB2 and 14-3-3zeta, with empty vector as a control. Cell motility assays showed that MECs overexpressing ErbB2 alone exhibited notably high migration speeds when cultured atop two dimensional (2D) matrices, while overexpression of 14-3-3zeta alone most suppressed migration atop 2D matrices (as compared to non-transformed MECs). Our results also suggest that co-overexpression of the 14-3-3zeta and ErbB2 proteins facilitates cell migratory capacity in 3D matrices, as reflected in cell migration speed. Additionally, 3D matrices of sufficient stiffness can significantly hinder the migratory ability of partially transformed cells, but increased 3D matrix stiffness has a lesser effect on the aggressive migratory behavior exhibited by fully transformed cells that co-overexpress both ErbB2 and 14-3-3zeta. Finally, this study shows that for MECs possessing partial or full transforming potential, those overexpressing ErbB2 alone show the greatest sensitivity of cell migration speed to matrix architecture, while those overexpressing 14-3-3zeta alone exhibit the least sensitivity to matrix architecture. Given the current knowledge of breast cancer mechanobiology, these findings overall suggest that cell motility is governed by a complex interplay between matrix mechanics and transforming potential. PMID- 21647372 TI - Pharmacogenetic modulation of orexin neurons alters sleep/wakefulness states in mice. AB - Hypothalamic neurons expressing neuropeptide orexins are critically involved in the control of sleep and wakefulness. Although the activity of orexin neurons is thought to be influenced by various neuronal input as well as humoral factors, the direct consequences of changes in the activity of these neurons in an intact animal are largely unknown. We therefore examined the effects of orexin neuron specific pharmacogenetic modulation in vivo by a new method called the Designer Receptors Exclusively Activated by Designer Drugs approach (DREADD). Using this system, we successfully activated and suppressed orexin neurons as measured by Fos staining. EEG and EMG recordings suggested that excitation of orexin neurons significantly increased the amount of time spent in wakefulness and decreased both non-rapid eye movement (NREM) and rapid eye movement (REM) sleep times. Inhibition of orexin neurons decreased wakefulness time and increased NREM sleep time. These findings clearly show that changes in the activity of orexin neurons can alter the behavioral state of animals and also validate this novel approach for manipulating neuronal activity in awake, freely-moving animals. PMID- 21647373 TI - Rapid reactivation of extralymphoid CD4 T cells during secondary infection. AB - After infection, extralymphoid tissues are enriched with effector and memory T cells of a highly activated phenotype. The capacity for rapid effector cytokine response from extralymphoid tissue-memory T cells suggests these cells may perform a 'sentinel' function in the tissue. While it has been demonstrated that extralymphoid CD4+ T cells can directly respond to secondary infection, little is known about how rapidly this response is initiated, and how early activation of T cells in the tissue may affect the innate response to infection. Here we use a mouse model of secondary heterosubtypic influenza infection to show that CD4(+) T cells in the lung airways are reactivated within 24 hours of secondary challenge. Airway CD4(+) T cells initiate an inflammatory cytokine and chemokine program that both alters the composition of the early innate response and contributes to the reduction of viral titers in the lung. These results show that, unlike a primary infection, extralymphoid tissue-memory CD4(+) T cells respond alongside the innate response during secondary infection, thereby shaping the overall immune profile in the airways. These data provide new insights into the role of extralymphoid CD4(+) T cells during secondary immune responses. PMID- 21647374 TI - Markov models of amino acid substitution to study proteins with intrinsically disordered regions. AB - BACKGROUND: Intrinsically disordered proteins (IDPs) or proteins with disordered regions (IDRs) do not have a well-defined tertiary structure, but perform a multitude of functions, often relying on their native disorder to achieve the binding flexibility through changing to alternative conformations. Intrinsic disorder is frequently found in all three kingdoms of life, and may occur in short stretches or span whole proteins. To date most studies contrasting the differences between ordered and disordered proteins focused on simple summary statistics. Here, we propose an evolutionary approach to study IDPs, and contrast patterns specific to ordered protein regions and the corresponding IDRs. RESULTS: Two empirical Markov models of amino acid substitutions were estimated, based on a large set of multiple sequence alignments with experimentally verified annotations of disordered regions from the DisProt database of IDPs. We applied new methods to detect differences in Markovian evolution and evolutionary rates between IDRs and the corresponding ordered protein regions. Further, we investigated the distribution of IDPs among functional categories, biochemical pathways and their preponderance to contain tandem repeats. CONCLUSIONS: We find significant differences in the evolution between ordered and disordered regions of proteins. Most importantly we find that disorder promoting amino acids are more conserved in IDRs, indicating that in some cases not only amino acid composition but the specific sequence is important for function. This conjecture is also reinforced by the observation that for of our data set IDRs evolve more slowly than the ordered parts of the proteins, while we still support the common view that IDRs in general evolve more quickly. The improvement in model fit indicates a possible improvement for various types of analyses e.g. de novo disorder prediction using a phylogenetic Hidden Markov Model based on our matrices showed a performance similar to other disorder predictors. PMID- 21647375 TI - Plasma cytokine profiles in subjects with high-functioning autism spectrum disorders. AB - BACKGROUND: Accumulating evidence suggests that dysregulation of the immune system is involved in the pathophysiology of autism spectrum disorders (ASD). The aim of the study was to explore immunological markers in peripheral plasma samples from non-medicated subjects with high-functioning ASD. METHODOLOGY/PRINCIPAL FINDINGS: A multiplex assay for cytokines and chemokines was applied to plasma samples from male subjects with high-functioning ASD (n = 28) and matched controls (n = 28). Among a total of 48 analytes examined, the plasma concentrations of IL-1beta, IL-1RA, IL-5, IL-8, IL-12(p70), IL-13, IL-17 and GRO-alpha were significantly higher in subjects with ASD compared with the corresponding values of matched controls after correction for multiple comparisons. CONCLUSION/SIGNIFICANCE: The results suggest that abnormal immune responses as assessed by multiplex analysis of cytokines may serve as one of the biological trait markers for ASD. PMID- 21647376 TI - Growth-rate dependence reveals design principles of plasmid copy number control. AB - Genetic circuits in bacteria are intimately coupled to the cellular growth rate as many parameters of gene expression are growth-rate dependent. Growth-rate dependence can be particularly pronounced for genes on plasmids; therefore the native regulatory systems of a plasmid such as its replication control system are characterized by growth-rate dependent parameters and regulator concentrations. This natural growth-rate dependent variation of regulator concentrations can be used for a quantitative analysis of the design of such regulatory systems. Here we analyze the growth-rate dependence of parameters of the copy number control system of ColE1-type plasmids in E. coli. This analysis allows us to infer the form of the control function and suggests that the Rom protein increases the sensitivity of control. PMID- 21647377 TI - Identification of miRs-143 and -145 that is associated with bone metastasis of prostate cancer and involved in the regulation of EMT. AB - The principal problem arising from prostate cancer (PCa) is its propensity to metastasize to bone. MicroRNAs (miRNAs) play a crucial role in many tumor metastases. The importance of miRNAs in bone metastasis of PCa has not been elucidated to date. We investigated whether the expression of certain miRNAs was associated with bone metastasis of PCa. We examined the miRNA expression profiles of 6 primary and 7 bone metastatic PCa samples by miRNA microarray analysis. The expression of 5 miRNAs significantly decreased in bone metastasis compared with primary PCa, including miRs-508-5p, -145, -143, -33a and -100. We further examined other samples of 16 primary PCa and 13 bone metastases using real-time PCR analysis. The expressions of miRs-143 and -145 were verified to down-regulate significantly in metastasis samples. By investigating relationship of the levels of miRs-143 and -145 with clinicopathological features of PCa patients, we found down-regulations of miRs-143 and -145 were negatively correlated to bone metastasis, the Gleason score and level of free PSA in primary PCa. Over expression miR-143 and -145 by retrovirus transfection reduced the ability of migration and invasion in vitro, and tumor development and bone invasion in vivo of PC-3 cells, a human PCa cell line originated from a bone metastatic PCa specimen. Their upregulation also increased E-cadherin expression and reduced fibronectin expression of PC-3 cells which revealed a less invasive morphologic phenotype. These findings indicate that miRs-143 and -145 are associated with bone metastasis of PCa and suggest that they may play important roles in the bone metastasis and be involved in the regulation of EMT Both of them may also be clinically used as novel biomarkers in discriminating different stages of human PCa and predicting bone metastasis. PMID- 21647378 TI - Common peptides study of aminoacyl-tRNA synthetases. AB - BACKGROUND: Aminoacyl tRNA synthetases (aaRSs) constitute an essential enzyme super-family, providing fidelity of the translation process of mRNA to proteins in living cells. They are common to all kingdoms and are of utmost importance to all organisms. It is thus of great interest to understand the evolutionary relationships among them and underline signature motifs defining their common domains. RESULTS: We utilized the Common Peptides (CPs) framework, based on extracted deterministic motifs from all aaRSs, to study family-specific properties. We identified novel aaRS-class related signatures that may supplement the current classification methods and provide a basis for identifying functional regions specific to each aaRS class. We exploited the space spanned by the CPs in order to identify similarities between aaRS families that are not observed using sequence alignment methods, identifying different inter-aaRS associations across different kingdom of life. We explored the evolutionary history of the aaRS families and evolutionary origins of the mitochondrial aaRSs. Lastly, we showed that prevalent CPs significantly overlap known catalytic and binding sites, suggesting that they have meaningful functional roles, as well as identifying a motif shared between aaRSs and a the Biotin-[acetyl-CoA carboxylase] synthetase (birA) enzyme overlapping binding sites in both families. CONCLUSIONS: The study presents the multitude of ways to exploit the CP framework in order to extract meaningful patterns from the aaRS super-family. Specific CPs, discovered in this study, may play important roles in the functionality of these enzymes. We explored the evolutionary patterns in each aaRS family and tracked remote evolutionary links between these families. PMID- 21647379 TI - Mitotic spindle proteomics in Chinese hamster ovary cells. AB - Mitosis is a fundamental process in the development of all organisms. The mitotic spindle guides the cell through mitosis as it mediates the segregation of chromosomes, the orientation of the cleavage furrow, and the progression of cell division. Birth defects and tissue-specific cancers often result from abnormalities in mitotic events. Here, we report a proteomic study of the mitotic spindle from Chinese Hamster Ovary (CHO) cells. Four different isolations of metaphase spindles were subjected to Multi-dimensional Protein Identification Technology (MudPIT) analysis and tandem mass spectrometry. We identified 1155 proteins and used Gene Ontology (GO) analysis to categorize proteins into cellular component groups. We then compared our data to the previously published CHO midbody proteome and identified proteins that are unique to the CHO spindle. Our data represent the first mitotic spindle proteome in CHO cells, which augments the list of mitotic spindle components from mammalian cells. PMID- 21647380 TI - Solution structure of LC4 transmembrane segment of CCR5. AB - CC-chemokine receptor 5 (CCR5) is a specific co-receptor allowing the entry of human immunodeficiency virus type 1 (HIV-1). The LC4 region in CCR5 is required for HIV-1 entry into the cells. In this study, the solution structure of LC4 in SDS micelles was elucidated by using standard 1H two-dimensional NMR spectroscopy, circular dichroism, and fluorescence quenching. The LC4 structure adopts two helical structures, whereas the C-terminal part remains unstructured. The positions in which LC4 binds to the HIV-1 inhibitory peptide LC5 were determined by docking calculations in addition to NMR data. The poses showed the importance of the hydrophobic interface of the assembled structures. The solution structure of LC4 elucidated in the present work provides a structural basis for further studies on the HIV-1 inhibitory function of the LC4 region. PMID- 21647381 TI - siRNA-like double-stranded RNAs are specifically protected against degradation in human cell extract. AB - RNA interference (RNAi) is a set of intracellular pathways in eukaryotes that controls both exogenous and endogenous gene expression. The power of RNAi to knock down (silence) any gene of interest by the introduction of synthetic small interfering (si)RNAs has afforded powerful insight into biological function through reverse genetic approaches and has borne a new field of gene therapeutics. A number of questions are outstanding concerning the potency of siRNAs, necessitating an understanding of how short double-stranded RNAs are processed by the cell. Recent work suggests unmodified siRNAs are protected in the intracellular environment, although the mechanism of protection still remains unclear. We have developed a set of doubly-fluorophore labeled RNAs (more precisely, RNA/DNA chimeras) to probe in real-time the stability of siRNAs and related molecules by fluorescence resonance energy transfer (FRET). We find that these RNA probes are substrates for relevant cellular degradative processes, including the RNase H1 mediated degradation of an DNA/RNA hybrid and Dicer mediated cleavage of a 24-nucleotide (per strand) double-stranded RNA. In addition, we find that 21- and 24-nucleotide double-stranded RNAs are relatively protected in human cytosolic cell extract, but less so in blood serum, whereas an 18-nucleotide double-stranded RNA is less protected in both fluids. These results suggest that RNAi effector RNAs are specifically protected in the cellular environment and may provide an explanation for recent results showing that unmodified siRNAs in cells persist intact for extended periods of time. PMID- 21647382 TI - An autotetraploid linkage map of rose (Rosa hybrida) validated using the strawberry (Fragaria vesca) genome sequence. AB - Polyploidy is a pivotal process in plant evolution as it increase gene redundancy and morphological intricacy but due to the complexity of polysomic inheritance we have only few genetic maps of autopolyploid organisms. A robust mapping framework is particularly important in polyploid crop species, rose included (2n = 4x = 28), where the objective is to study multiallelic interactions that control traits of value for plant breeding. From a cross between the garden, peach red and fragrant cultivar Fragrant Cloud (FC) and a cut-rose yellow cultivar Golden Gate (GG), we generated an autotetraploid GGFC mapping population consisting of 132 individuals. For the map we used 128 sequence-based markers, 141 AFLP, 86 SSR and three morphological markers. Seven linkage groups were resolved for FC (Total 632 cM) and GG (616 cM) which were validated by markers that segregated in both parents as well as the diploid integrated consensus map.The release of the Fragaria vesca genome, which also belongs to the Rosoideae, allowed us to place 70 rose sequenced markers on the seven strawberry pseudo-chromosomes. Synteny between Rosa and Fragaria was high with an estimated four major translocations and six inversions required to place the 17 non-collinear markers in the same order. Based on a verified linear order of the rose markers, we could further partition each of the parents into its four homologous groups, thus providing an essential framework to aid the sequencing of an autotetraploid genome. PMID- 21647383 TI - A simple index for the high-citation tail of citation distribution to quantify research performance in countries and institutions. AB - BACKGROUND: Conventional scientometric predictors of research performance such as the number of papers, citations, and papers in the top 1% of highly cited papers cannot be validated in terms of the number of Nobel Prize achievements across countries and institutions. The purpose of this paper is to find a bibliometric indicator that correlates with the number of Nobel Prize achievements. METHODOLOGY/PRINCIPAL FINDINGS: This study assumes that the high-citation tail of citation distribution holds most of the information about high scientific performance. Here I propose the x-index, which is calculated from the number of national articles in the top 1% and 0.1% of highly cited papers and has a subtractive term to discount highly cited papers that are not scientific breakthroughs. The x-index, the number of Nobel Prize achievements, and the number of national articles in Nature or Science are highly correlated. The high correlations among these independent parameters demonstrate that they are good measures of high scientific performance because scientific excellence is their only common characteristic. However, the x-index has superior features as compared to the other two parameters. Nobel Prize achievements are low frequency events and their number is an imprecise indicator, which in addition is zero in most institutions; the evaluation of research making use of the number of publications in prestigious journals is not advised. CONCLUSION: The x-index is a simple and precise indicator for high research performance. PMID- 21647384 TI - Asthma pregnancy alters postnatal development of chromaffin cells in the rat adrenal medulla. AB - BACKGROUND: Adrenal neuroendocrine plays an important role in asthma. The activity of the sympathoadrenal system could be altered by early life events. The effects of maternal asthma during pregnancy on the adrenal medulla of offspring remain unknown. METHODOLOGY/PRINCIPAL FINDINGS: This study aims to explore the influence of maternal asthma during pregnancy on the development and function of adrenal medulla in offspring from postnatal day 3 (P3) to postnatal day 60 (P60). Asthmatic pregnant rats (AP), nerve growth factor (NGF)-treated pregnant rats (NP) and NGF antibody-treated pregnant rats (ANP) were sensitized and challenged with ovalbumin (OVA); NP and ANP were treated with NGF and NGF antibody respectively. Offspring rats from the maternal group were divided into four groups: offspring from control pregnant rats (OCP), offspring from AP (OAP), offspring from NP (ONP), and offspring from ANP (OANP). The expressions of phenylethanolamine N-methyltransferase (PNMT) protein in adrenal medulla were analyzed. The concentrations of epinephrine (EPI), corticosterone and NGF in serum were measured. Adrenal medulla chromaffin cells (AMCC) were prone to differentiate into sympathetic nerve cells in OAP and ONP. Both EPI and PNMT were decreased in OAP from P3 to P14, and then reached normal level gradually from P30 to P60, which were lower from birth to adulthood in ONP. Corticosterone concentration increased significantly in OAP and ONP. CONCLUSION/SIGNIFICANCE: Asthma pregnancy may promote AMCC to differentiate into sympathetic neurons in offspring rats and inhibit the synthesis of EPI, resulting in dysfunction of bronchial relaxation. PMID- 21647385 TI - Control of mitochondrial morphology through differential interactions of mitochondrial fusion and fission proteins. AB - Mitochondria in mammals are organized into tubular networks that undergo frequent shape change. Mitochondrial fission and fusion are the main components mediating the mitochondrial shape change. Perturbation of the fission/fusion balance is associated with many disease conditions. However, underlying mechanisms of the fission/fusion balance are not well understood. Mitochondrial fission in mammals requires the dynamin-like protein DLP1/Drp1 that is recruited to the mitochondrial surface, possibly through the membrane-anchored protein Fis1 or Mff. Additional dynamin-related GTPases, mitofusin (Mfn) and OPA1, are associated with the outer and inner mitochondrial membranes, respectively, and mediate fusion of the respective membranes. In this study, we found that two heptad repeat regions (HR1 and HR2) of Mfn2 interact with each other, and that Mfn2 also interacts with the fission protein DLP1. The association of the two heptad repeats of Mfn2 is fusion inhibitory whereas a positive role of the Mfn2/DLP1 interaction in mitochondrial fusion is suggested. Our results imply that the differential binding of Mfn2-HR1 to HR2 and DLP1 regulates mitochondrial fusion and that DLP1 may act as a regulatory factor for efficient execution of both fusion and fission of mitochondria. PMID- 21647386 TI - Lysophosphatidate induces chemo-resistance by releasing breast cancer cells from taxol-induced mitotic arrest. AB - BACKGROUND: Taxol is a microtubule stabilizing agent that arrests cells in mitosis leading to cell death. Taxol is widely used to treat breast cancer, but resistance occurs in 25-69% of patients and it is vital to understand how Taxol resistance develops to improve chemotherapy. The effects of chemotherapeutic agents are overcome by survival signals that cancer cells receive. We focused our studies on autotaxin, which is a secreted protein that increases tumor growth, aggressiveness, angiogenesis and metastasis. We discovered that autotaxin strongly antagonizes the Taxol-induced killing of breast cancer and melanoma cells by converting the abundant extra-cellular lipid, lysophosphatidylcholine, into lysophosphatidate. This lipid stimulates specific G-protein coupled receptors that activate survival signals. METHODOLOGY/PRINCIPAL FINDINGS: In this study we determined the basis of these antagonistic actions of lysophosphatidate towards Taxol-induced G2/M arrest and cell death using cultured breast cancer cells. Lysophosphatidate does not antagonize Taxol action in MCF-7 cells by increasing Taxol metabolism or its expulsion through multi-drug resistance transporters. Lysophosphatidate does not lower the percentage of cells accumulating in G2/M by decreasing exit from S-phase or selective stimulation of cell death in G2/M. Instead, LPA had an unexpected and remarkable action in enabling MCF-7 and MDA-MB-468 cells, which had been arrested in G2/M by Taxol, to normalize spindle structure and divide, thus avoiding cell death. This action involves displacement of Taxol from the tubulin polymer fraction, which based on inhibitor studies, depends on activation of LPA receptors and phosphatidylinositol 3-kinase. CONCLUSIONS/SIGNIFICANCE: This work demonstrates a previously unknown consequence of lysophosphatidate action that explains why autotaxin and lysophosphatidate protect against Taxol-induced cell death and promote resistance to the action of this important therapeutic agent. PMID- 21647387 TI - Bicistronic lentiviruses containing a viral 2A cleavage sequence reliably co express two proteins and restore vision to an animal model of LCA1. AB - The disease processes underlying inherited retinal disease are complex and are not completely understood. Many of the corrective gene therapies designed to treat diseases linked to mutations in genes specifically expressed in photoreceptor cells restore function to these cells but fail to stop progression of the disease. There is growing consensus that effective treatments for these diseases will require delivery of multiple therapeutic proteins that will be selected to treat specific aspects of the disease process. The purpose of this study was to design a lentiviral transgene that reliably expresses all of the proteins it encodes and does so in a consistent manner among infected cells. We show, using both in vitro and in vivo analyses, that bicistronic lentiviral transgenes encoding two fluorescent proteins fused to a viral 2A-like cleavage peptide meet these expression criteria. To determine if this transgene design is suitable for therapeutic applications, we replaced one of the fluorescent protein genes with the gene encoding guanylate cyclase-1 (GC1) and delivered lentivirus carrying this transgene to the retinas of the GUCY1*B avian model of Leber congenital amaurosis-1 (LCA1). GUCY1*B chickens carry a null mutation in the GC1 gene that disrupts photoreceptor function and causes blindness at hatching, a phenotype that closely matches that observed in humans with LCA1. We found that treatment of these animals with the 2A lentivector encoding GC1 restored vision to these animals as evidenced by the presence of optokinetic reflexes. We conclude that 2A-like peptides, with proper optimization, can be successfully incorporated into therapeutic vectors designed to deliver multiple proteins to neural retinal. These results highlight the potential of this vector design to serve as a platform for the development of combination therapies designed to enhance or prolong the benefits of corrective gene therapies. PMID- 21647389 TI - Correlation and cause when inferring attentional guidance in the rainforest and beyond. PMID- 21647388 TI - Estimating the threshold surface density of Gp120-CCR5 complexes necessary for HIV-1 envelope-mediated cell-cell fusion. AB - Reduced expression of CCR5 on target CD4(+) cells lowers their susceptibility to infection by R5-tropic HIV-1, potentially preventing transmission of infection and delaying disease progression. Binding of the HIV-1 envelope (Env) protein gp120 with CCR5 is essential for the entry of R5 viruses into target cells. The threshold surface density of gp120-CCR5 complexes that enables HIV-1 entry remains poorly estimated. We constructed a mathematical model that mimics Env mediated cell-cell fusion assays, where target CD4(+)CCR5(+) cells are exposed to effector cells expressing Env in the presence of a coreceptor antagonist and the fraction of target cells fused with effector cells is measured. Our model employs a reaction network-based approach to describe protein interactions that precede viral entry coupled with the ternary complex model to quantify the allosteric interactions of the coreceptor antagonist and predicts the fraction of target cells fused. By fitting model predictions to published data of cell-cell fusion in the presence of the CCR5 antagonist vicriviroc, we estimated the threshold surface density of gp120-CCR5 complexes for cell-cell fusion as ~20 um(-2). Model predictions with this threshold captured data from independent cell-cell fusion assays in the presence of vicriviroc and rapamycin, a drug that modulates CCR5 expression, as well as assays in the presence of maraviroc, another CCR5 antagonist, using sixteen different Env clones derived from transmitted or early founder viruses. Our estimate of the threshold surface density of gp120-CCR5 complexes necessary for HIV-1 entry thus appears robust and may have implications for optimizing treatment with coreceptor antagonists, understanding the non pathogenic infection of non-human primates, and designing vaccines that suppress the availability of target CD4(+)CCR5(+) cells. PMID- 21647390 TI - Laparoscopic-Assisted Cryptorchidectomy in an Adult Reindeer (Rangifer tarandus). AB - A successful laparoscopic-assisted cryptorchidectomy is reported in a novel species, the reindeer (Rangifer tarandus). The procedure was performed in an 8 year-old adult positioned in dorsolateral oblique recumbency, with an open approach midline subumbilical placement of the primary 10 mm optical port and carbon dioxide insufflation at 12 mmHg. Three 5 mm instrument ports were inserted under visualization in the left caudal abdomen as the retained testicle was localized to the internal inguinal ring. A 5 mm flexible organ retractor was used to assist in localizing the retained testicle. This procedure provided a less invasive alternative to open laparotomy. The authors are unaware of any published reports of laparoscopy in reindeer, or of laparoscopic assisted cryptorchidectomy in deer species. PMID- 21647391 TI - Comparative gamma delta T cell immunology: a focus on mycobacterial disease in cattle. AB - A theme among many pathogenic mycobacterial species affecting both humans and animals is a prolonged asymptomatic or latent period that can last years to decades. The mechanisms that favor progression to active disease are not well understood. Pathogen containment is often associated with an effective cell mediated or T-helper 1 immune profile. With certain pathogenic mycobacteria, such as Mycobacterium avium subspecies paratuberculosis, a shift to active clinical disease is associated with loss of T-helper 1 immunity and development of an ineffective humoral or T-helper 2 immune response. Recently gammadelta T cells have been shown to play a role early in mycobacterial infections and have been hypothesized to influence disease outcome. The purpose of this paper is to compare recent advancements in our understanding of gammadelta T cells in humans, cattle, and mice and to discuss roles of gammadelta T cells in host response to mycobacterial infection. PMID- 21647392 TI - Intrasulcal electrocorticography in macaque monkeys with minimally invasive neurosurgical protocols. AB - Electrocorticography (ECoG), multichannel brain-surface recording and stimulation with probe electrode arrays, has become a potent methodology not only for clinical neurosurgery but also for basic neuroscience using animal models. The highly evolved primate's brain has deep cerebral sulci, and both gyral and intrasulcal cortical regions have been implicated in important functional processes. However, direct experimental access is typically limited to gyral regions, since placing probes into sulci is difficult without damaging the surrounding tissues. Here we describe a novel methodology for intrasulcal ECoG in macaque monkeys. We designed and fabricated ultra-thin flexible probes for macaques with micro-electro-mechanical systems technology. We developed minimally invasive operative protocols to implant the probes by introducing cutting-edge devices for human neurosurgery. To evaluate the feasibility of intrasulcal ECoG, we conducted electrophysiological recording and stimulation experiments. First, we inserted parts of the Parylene-C-based probe into the superior temporal sulcus to compare visually evoked ECoG responses from the ventral bank of the sulcus with those from the surface of the inferior temporal cortex. Analyses of power spectral density and signal-to-noise ratio revealed that the quality of the ECoG signal was comparable inside and outside of the sulcus. Histological examination revealed no obvious physical damage in the implanted areas. Second, we placed a modified silicone ECoG probe into the central sulcus and also on the surface of the precentral gyrus for stimulation. Thresholds for muscle twitching were significantly lower during intrasulcal stimulation compared to gyral stimulation. These results demonstrate the feasibility of intrasulcal ECoG in macaques. The novel methodology proposed here opens up a new frontier in neuroscience research, enabling the direct measurement and manipulation of electrical activity in the whole brain. PMID- 21647393 TI - Predictors of clients' satisfaction with delivery of animal health care services in periurban ghana. AB - The study used logistic regression modelling to determine predictors of satisfaction with delivery of animal health care services for 889 clients (livestock and poultry keepers) in periurban Ghana. Of the 15 indicators tested as predictors of satisfaction in this study, 8 were included in the best fit model. These were accessibility, availability of services, service charge, effectiveness, efficiency, quality of services, meeting client needs, and getting help. Efficiency and effectiveness were perceived by the respondents to be synonymous, as were service quality and effectiveness, as suggested by ORs > 10 when cross tabulated. Therefore, one or the other could be used in future studies but not both to avoid collinearity. The identified predictors could be targeted for improvement in quality of service delivery to livestock and poultry keepers in Ghana. PMID- 21647394 TI - Illthrift in suckling lambs secondary to umbilical infections and possible implication of cryptosporidiosis as a risk factor. AB - Illthrift was observed in 20/60 lambs aged 40-45 days in a dairy sheep flock in Greece. Cryptosporidiosis had been diagnosed and successfully treated with halofuginone lactate a month earlier. Parasitological examinations were negative for endoparasites while hematology and biochemistry were unremarkable. Necropsy of 5 lambs revealed lung and liver abscessation, presumably secondary to umbilical infections due to poor farm hygiene, though umbilical lesions were not observed. No new cases were observed following treatment of the umbilicus of newborn lambs with chlorexidine. Although umbilical infections are common, this is the first reported case of illthrift in lambs attributed to umbilical infection; illthrift may be the only clinical manifestation of such infections. The prior presence of cryptosporidiosis may have contributed to the severity of the infection through the reduction of local immunity. Recognition of this possibly underdiagnosed or underappreciated condition may improve medical, production, and welfare standards in the sheep industry. PMID- 21647395 TI - Regulation of the Fear Network by Mediators of Stress: Norepinephrine Alters the Balance between Cortical and Subcortical Afferent Excitation of the Lateral Amygdala. AB - Pavlovian auditory fear conditioning involves the integration of information about an acoustic conditioned stimulus (CS) and an aversive unconditioned stimulus in the lateral nucleus of the amygdala (LA). The auditory CS reaches the LA subcortically via a direct connection from the auditory thalamus and also from the auditory association cortex itself. How neural modulators, especially those activated during stress, such as norepinephrine (NE), regulate synaptic transmission and plasticity in this network is poorly understood. Here we show that NE inhibits synaptic transmission in both the subcortical and cortical input pathway but that sensory processing is biased toward the subcortical pathway. In addition binding of NE to beta-adrenergic receptors further dissociates sensory processing in the LA. These findings suggest a network mechanism that shifts sensory balance toward the faster but more primitive subcortical input. PMID- 21647397 TI - The microcircuit concept applied to cortical evolution: from three-layer to six layer cortex. AB - Understanding the principles of organization of the cerebral cortex requires insight into its evolutionary history. This has traditionally been the province of anatomists, but evidence regarding the microcircuit organization of different cortical areas is providing new approaches to this problem. Here we use the microcircuit concept to focus first on the principles of microcircuit organization of three-layer cortex in the olfactory cortex, hippocampus, and turtle general cortex, and compare it with six-layer neocortex. From this perspective it is possible to identify basic circuit elements for recurrent excitation and lateral inhibition that are common across all the cortical regions. Special properties of the apical dendrites of pyramidal cells are reviewed that reflect the specific adaptations that characterize the functional operations in the different regions. These principles of microcircuit function provide a new approach to understanding the expanded functional capabilities elaborated by the evolution of the neocortex. PMID- 21647398 TI - Authenticity, depression, and deep brain stimulation. PMID- 21647396 TI - Long-Term Potentiation at CA3-CA1 Hippocampal Synapses with Special Emphasis on Aging, Disease, and Stress. AB - Synaptic plasticity in the mammalian central nervous system has been the subject of intense investigation for the past four decades. Long-term potentiation (LTP), a major reflection of synaptic plasticity, is an activity-driven long-lasting increase in the efficacy of excitatory synaptic transmission following the delivery of a brief, high-frequency train of electrical stimulation. LTP is regarded as a principal candidate for the cellular mechanisms involved in learning and offers an attractive hypothesis of how memories are constructed. There are a number of exceptional full-length reviews published on LTP; the current review intends to present an overview of the research findings regarding hippocampal LTP with special emphasis on aging, diseases, and psychological insults. PMID- 21647399 TI - The hippocampal neuroproteome with aging and cognitive decline: past progress and future directions. AB - Although steady progress on understanding brain aging has been made over recent decades through standard anatomical, immunohistochemical, and biochemical techniques, the biological basis of non-neurodegenerative cognitive decline with aging remains to be determined. This is due in part to technical limitations of traditional approaches, in which only a small fraction of neurobiologically relevant proteins, mRNAs or metabolites can be assessed at a time. With the development and refinement of proteomic technologies that enable simultaneous quantitative assessment of hundreds to thousands of proteins, neuroproteomic studies of brain aging and cognitive decline are becoming more widespread. This review focuses on the contributions of neuroproteomic investigations to advances in our understanding of age-related deficits of hippocampus-dependent spatial learning and memory. Accumulating neuroproteomic data demonstrate that hippocampal aging involves common themes of dysregulated metabolism, increased oxidative stress, altered protein processing, and decreased synaptic function. Additionally, growing evidence suggests that cognitive decline does not represent a "more aged" phenotype, but rather is associated with specific neuroproteomic changes that occur in addition to age-related alterations. Understanding if and how age-related changes in the hippocampal neuroproteome contribute to cognitive decline and elucidating the pathways and processes that lead to cognitive decline are critical objectives that remain to be achieved. Progress in the field and challenges that remain to be addressed with regard to animal models, behavioral testing, and proteomic reporting are also discussed. PMID- 21647401 TI - Antigen-based immune therapeutics for type 1 diabetes: magic bullets or ordinary blanks? AB - The ideal drug of modern medicine is the one that achieves its therapeutic target with minimal adverse effects. Immune therapy of Type 1 diabetes (T1D) is no exception, and knowledge of the antigens targeted by pathogenic T cells offers a unique opportunity towards this goal. Different antigen formulations are being considered, such as proteins or peptides, either in their native form or modified ad hoc, DNA plasmids, and cell-based agents. Translation from mouse to human should take into account important differences, particularly in the time scale of autoimmune progression, and intervention. Critical parameters such as administration route, dosing and interval remain largely empirical and need to be further dissected. T1D staging through immune surrogate markers before and after treatment will be key in understanding therapeutic actions and to finally turn ordinary blanks into magic bullets. PMID- 21647402 TI - Epigenetics of early child development. AB - Comprehensive clinical studies show that adverse conditions in early life can severely impact the developing brain and increase vulnerability to mood disorders later in life. During early postnatal life the brain exhibits high plasticity which allows environmental signals to alter the trajectories of rapidly developing circuits. Adversity in early life is able to shape the experience dependent maturation of stress-regulating pathways underlying emotional functions and endocrine responses to stress, such as the hypothalamo-pituitary-adrenal (HPA) system, leading to long-lasting altered stress responsivity during adulthood. To date, the study of gene-environment interactions in the human population has been dominated by epidemiology. However, recent research in the neuroscience field is now advancing clinical studies by addressing specifically the mechanisms by which gene-environment interactions can predispose individuals toward psychopathology. To this end, appropriate animal models are being developed in which early environmental factors can be manipulated in a controlled manner. Here we will review recent studies performed with the common aim of understanding the effects of the early environment in shaping brain development and discuss the newly developing role of epigenetic mechanisms in translating early life conditions into long-lasting changes in gene expression underpinning brain functions. Particularly, we argue that epigenetic mechanisms can mediate the gene-environment dialog in early life and give rise to persistent epigenetic programming of adult physiology and dysfunction eventually resulting in disease. Understanding how early life experiences can give rise to lasting epigenetic marks conferring increased risk for mental disorders, how they are maintained and how they could be reversed, is increasingly becoming a focus of modern psychiatry and should pave new guidelines for timely therapeutic interventions. PMID- 21647403 TI - Type I diabetes-associated tolerogenic properties of interleukin-2. AB - Type 1 Diabetes (T1D) results from insulin-producing beta cells destruction by diabetogenic T lymphocytes in humans and nonobese diabetic (NOD) mice. The breakdown of tolerance has been associated with a defect in the number and the function of naturally occurring regulatory T cells (nTreg) that are the master player in peripheral tolerance. Gene knockout experiments in mouse models have shown a nonredundant activity of IL-2 related to its critical role in inducing nTreg and controlling peripheral T cell tolerance. Whereas strong evidence has suggested that IL-2 is critically required for nTreg-mediated T1D control, several fundamental questions remain to be addressed. In this paper, we highlight the recent findings and controversies regarding the tolerogenic properties of IL 2 mediated through nTreg. We further discuss a potential link between the immunomodulatory role of interleukin-2 and the pathogenesis of type 1 diabetes. PMID- 21647404 TI - Possible effects of synaptic imbalances on oligodendrocyte-axonic interactions in schizophrenia: a hypothetical model. AB - A model of glial-neuronal interactions is proposed that could be explanatory for the demyelination identified in brains with schizophrenia. It is based on two hypotheses: (1) that glia-neuron systems are functionally viable and important for normal brain function, and (2) that disruption of this postulated function disturbs the glial categorization function, as shown by formal analysis. According to this model, in schizophrenia receptors on astrocytes in glial neuronal synaptic units are not functional, loosing their modulatory influence on synaptic neurotransmission. Hence, an unconstrained neurotransmission flux occurs that hyperactivates the axon and floods the cognate receptors of neurotransmitters on oligodendrocytes. The excess of neurotransmitters may have a toxic effect on oligodendrocytes and myelin, causing demyelination. In parallel, an increasing impairment of axons may disconnect neuronal networks. It is formally shown how oligodendrocytes normally categorize axonic information processing via their processes. Demyelination decomposes the oligodendrocyte axonic system making it incapable to generate categories of information. This incoherence may be responsible for symptoms of disorganization in schizophrenia, such as thought disorder, inappropriate affect and incommunicable motor behavior. In parallel, the loss of oligodendrocytes affects gap junctions in the panglial syncytium, presumably responsible for memory impairment in schizophrenia. PMID- 21647405 TI - Thymic self-antigen expression for the design of a negative/tolerogenic self vaccine against type 1 diabetes. AB - Before being able to react against infectious non-self-antigens, the immune system has to be educated in the recognition and tolerance of neuroendocrine proteins, and this critical process essentially takes place in the thymus. The development of the autoimmune diabetogenic response results from a thymus dysfunction in programming central self-tolerance to pancreatic insulin-secreting islet beta cells, leading to the breakdown of immune homeostasis with an enrichment of islet beta cell reactive effector T cells and a deficiency of beta cell-specific natural regulatory T cells (nTreg) in the peripheral T-lymphocyte repertoire. Insulin-like growth factor 2 (IGF-2) is the dominant member of the insulin family expressed during fetal life by the thymic epithelium under the control of the autoimmune regulator (AIRE) gene/protein. Based on the close homology and cross-tolerance between insulin, the primary T1D autoantigen, and IGF-2, the dominant self-antigen of the insulin family, a novel type of vaccination, so-called "negative/tolerogenic self-vaccination", is currently developed for prevention and cure of T1D. If this approach were found to be effective for reprogramming immunological tolerance in T1D, it could pave the way for the design of negative self-vaccines against autoimmune endocrine diseases, as well as other organ-specific autoimmune diseases. PMID- 21647406 TI - IFN regulatory factors 4 and 8 expression in the NOD mouse. AB - Dendritic cells (DCs) contribute to islet inflammation and its progression to diabetes in NOD mouse model and human. DCs play a crucial role in the presentation of autoantigen and activation of diabetogenic T cells, and IRF4 and IRF8 are crucial genes involved in the development of DCs. We have therefore investigated the expression of these genes in splenic DCs during diabetes progression in NOD mice. We found that IRF4 expression was upregulated in splenocytes and in splenic CD11c(+) DCs of NOD mice as compared to BALB/c mice. In contrast, IRF8 gene expression was higher in splenocytes of NOD mice whereas its expression was similar in splenic CD11c(+) DCs of NOD and BALB/c mice. Importantly, levels of IRF4 and IRF8 expression were lower in tolerogenic bone marrow derived DCs (BMDCs) generated with GM-CSF as compared to immunogenic BMDCs generated with GM-CSF and IL-4. Analysis of splenic DCs subsets indicated that high expression of IRF4 was associated with increased levels of CD4(+)CD8alpha( )IRF4(+)CD11c(+) DCs but not CD4(-)CD8alpha(+)IRF8(+)CD11c(+) DCs in NOD mice. Our results showed that IRF4 expression was up-regulated in NOD mice and correlated with the increased levels of CD4(+)CD8alpha(-) DCs, suggesting that IRF4 may be involved in abnormal DC functions in type 1 diabetes in NOD mice. PMID- 21647407 TI - The role of CXCR3 in the induction of primary biliary cirrhosis. AB - OBJECTIVE: Investigate whether CXCR3 and its ligands were involved in the pathogenesis of primary biliary cirrhosis (PBC) in an autoimmune cholangitis animal model. METHODS: Female C57BL/6 mice were injected with 5 mg/kg of poly I:C intraperitoneally twice a week for 24 weeks. PBC model was confirmed by liver function, serum autoantibodies and liver biopsy. Lymphocytes subsets in liver and spleen and CXCL10 serum level were tested by flow cytometry and ELISA. Liver specimens were collected to evaluate the differences in pathology between WT and CXCR3(-/-) mice. RESULTS: Antimitochondrial antibody was detected in all PBC model. Numbers of infiltrates were detected in the portal areas 8 weeks after poly I:C injection, which progressed up to 24 weeks. Compared to control mice, CXCL10 serum level increased in PBC mice and the proportion of CXCR3(+) cells increased in the intrahepatic infiltrates of PBC mice, chiefly on CD8(+) cells, whereas the expression of CXCR3 on CD3(+) and CD8(+) splenocytes decreased in PBC model. Compared with WT mice, CXCR3(-/-) mice developed delayed and milder progression of cellular inflammation. CONCLUSIONS: CXCR3 might contribute to the development of PBC in murine model. Knockout of CXCR3 might delay and alleviate the PBC disease progression, but could not entirely block the disease development. PMID- 21647408 TI - Innate immune sensors and gastrointestinal bacterial infections. AB - The gastrointestinal microbiota is a major source of immune stimulation. The interaction between host pattern-recognition receptors and conserved microbial ligands profoundly influences infection dynamics. Identifying and understanding the nature of these interactions is a key step towards obtaining a clearer picture of microbial pathogenesis. These interactions underpin a complex interplay between microbe and host that has far reaching consequences for both. Here, we review the role of pattern recognition receptors in three prototype diseases affecting the stomach, the small intestine, and large intestine, respectively (Helicobacter pylori infection, Salmonella infection, and inflammatory bowel disease). Specifically, we review the nature and impact of pathogen:receptor interactions, their impact upon pathogenesis, and address the relevance of pattern recognition receptors in the development of therapies for gastrointestinal diseases. PMID- 21647409 TI - Oxidative stress and redox modulation potential in type 1 diabetes. AB - Redox reactions are imperative to preserving cellular metabolism yet must be strictly regulated. Imbalances between reactive oxygen species (ROS) and antioxidants can initiate oxidative stress, which without proper resolve, can manifest into disease. In type 1 diabetes (T1D), T-cell-mediated autoimmune destruction of pancreatic beta-cells is secondary to the primary invasion of macrophages and dendritic cells (DCs) into the islets. Macrophages/DCs, however, are activated by intercellular ROS from resident pancreatic phagocytes and intracellular ROS formed after receptor-ligand interactions via redox-dependent transcription factors such as NF-kappaB. Activated macrophages/DCs ferry beta cell antigens specifically to pancreatic lymph nodes, where they trigger reactive T cells through synapse formation and secretion of proinflammatory cytokines and more ROS. ROS generation, therefore, is pivotal in formulating both innate and adaptive immune responses accountable for islet cell autoimmunity. The importance of ROS/oxidative stress as well as potential for redox modulation in the context of T1D will be discussed. PMID- 21647410 TI - Recombinant HBHA boosting effect on BCG-induced immunity against Mycobacterium tuberculosis infection. AB - Heterologous prime-boost regimens are effective strategies to promote long-term memory and strong cellular Th1 responses to Mycobacterium tuberculosis, when BCG is used in the priming step. Subcutaneous or intranasal boosting of BCG vaccinated newborn mice with native heparin-binding haemagglutinin (nHBHA) significantly enhances protection against M. tuberculosis. However, nHBHA is characterized by a complex methylation pattern in its C-terminal domain, which is important for protective immunogenicity in primary vaccination. In this study we addressed the question whether boosting with recombinant, non-methylated HBHA (rHBHA) produced in Escherichia coli may enhance protection of BCG-primed newborn mice. We found that while subcutaneous rHBHA boosting enhanced protection of BCG primed mice against intranasal M. tuberculosis infection both in spleen and lungs, enhanced protection against aerosol infection was only seen in the spleen (0.72 logs; P < 0.05) but not in the lungs. Thus, in BCG-primed mice the methylation of the C-terminal domain of HBHA is dispensable for the induction of enhanced protection in the lungs against intranasal but not aerosol infection, whereas it enhances protection in the spleen in both challenge models. This report thus provides evidence that rHBHA may be considered as a booster vaccine against disseminated tuberculosis. PMID- 21647411 TI - Interferon alpha on expression of hTERT mRNA in peripheral blood mononuclear cells of patients with chronic hepatitis B. AB - Cell division is closely related to telomerase activity (hTERT mRNA). Lower expression of lymphocitic hTERT mRNA may easily cause cell aging, which is not beneficial to maintaining a durable lymphocyte division. To date, there is no study to investigate IFNalpha therapy on hTERT mRNA expression in PBMCs of patients with chronic hepatitis B (CHB). We quantitatively detected hTERT mRNA from study subjects and made each hTERT mRNA normalized (NhTERT mRNA). Mean NhTERT mRNA level was lower in either CHB group, but it significantly increased in IFNalpha-treated group compared with CHB control group, and a longer duration of IFNalpha therapy could increase the level. Moreover, the mean NhTERT mRNA in subgroup with HBeAg loss was significantly higher than that in subgroup without. NhTERT mRNA was markedly correlated with CD3(+) T lymphocyte count and CD4(+)/CD8(+) ratio. The results showed that IFNalpha therapy could upregulate the expression of hTERT mRNA in PBMCs. PMID- 21647412 TI - Deep brain stimulation for Parkinson's disease: where do we stand? PMID- 21647413 TI - The Smell of Blue Light: A New Approach toward Understanding an Olfactory Neuronal Network. AB - Olfaction is one of the most important senses throughout the animal kingdom. It enables animals to discriminate between a wide variety of attractive and repulsive odorants and often plays a decisive role in species specific communication. In recent years the analysis of olfactory systems both invertebrates and invertebrates has attracted much scientific interest. In this context a pivotal question is how the properties and connectivities of individual neurons contribute to a functioning neuronal network that mediates odor-guided behavior. As a novel approach to analyze the role of individual neurons within a circuitry, techniques have been established that make use of light-sensitive proteins. In this review we introduce a non-invasive, optogenetic technique which was used to manipulate the activity of individual neurons in the olfactory system of Drosophila melanogaster larvae. Both channelrhodopsin-2 and the photosensitive adenylyl cyclase PAC alpha in individual olfactory receptor neurons (ORNs) of the olfactory system of Drosophila larvae allows stimulating individual receptor neurons by light. Depending on which particular ORN is optogenetically activated, repulsion or attraction behavior can be induced, indicating which sensory neurons underlie which type of behavior. PMID- 21647414 TI - Relational Dynamics in Perception: Impacts on Trial-to-trial Variation. AB - We show that trial-to-trial variability in sensory detection of a weak visual stimulus is dramatically diminished when rather than presenting a fixed stimulus contrast, fluctuations in a subject's judgment are matched by fluctuations in stimulus contrast. This attenuation of fluctuations does not involve a change in the subject's psychometric function. The result is consistent with the interpretation of trial-to-trial variability in this sensory detection task being a high-level meta-cognitive control process that explores for something that our brains are so used to: subject-object relational dynamics. PMID- 21647415 TI - Crosstalk between medulloblastoma cells and endothelium triggers a strong chemotactic signal recruiting T lymphocytes to the tumor microenvironment. AB - Cancer cells can live and grow if they succeed in creating a favorable niche that often includes elements from the immune system. While T lymphocytes play an important role in the host response to tumor growth, the mechanism of their trafficking to the tumor remains poorly understood. We show here that T lymphocytes consistently infiltrate the primary brain cancer, medulloblastoma. We demonstrate, both in vitro and in vivo, that these T lymphocytes are attracted to tumor deposits only after the tumor cells have interacted with tumor vascular endothelium. Macrophage Migration Inhibitory Factor (MIF)" is the key chemokine molecule secreted by tumor cells which induces the tumor vascular endothelial cells to secrete the potent T lymphocyte attractant "Regulated upon Activation, Normal T-cell Expressed, and Secreted (RANTES)." This in turn creates a chemotactic gradient for RANTES-receptor bearing T lymphocytes. Manipulation of this pathway could have important therapeutic implications. PMID- 21647417 TI - Spontaneous firings of carnivorous aquatic Utricularia traps: temporal patterns and mechanical oscillations. AB - Aquatic species of Utricularia are carnivorous plants living in environments poor in nutrients. Their trapping mechanism has fascinated generations of scientists and is still debated today. It was reported recently that Utricularia traps can fire spontaneously. We show here that these spontaneous firings follow an unexpected diversity of temporal patterns, from "metronomic" traps which fire at fixed time intervals to "random" patterns, displaying more scattered firing times. Some "bursting" traps even combine both aspects, with groups of fast regular firings separated by a variable amount of time. We propose a physical model to understand these very particular behaviors, showing that a trap of Utricularia accomplishes mechanical oscillations, based on continuous pumping and sudden opening of the trap door (buckling). We isolate the key parameters governing these oscillations and discuss the effect of their fluctuations. PMID- 21647416 TI - Induction of macrophage chemotaxis by aortic extracts from patients with Marfan syndrome is related to elastin binding protein. AB - Marfan syndrome is an autosomal dominantly inherited disorder of connective tissue with prominent skeletal, ocular, and cardiovascular manifestations. Aortic aneurysm and dissection are the major determinants of premature death in untreated patients. In previous work, we showed that extracts of aortic tissues from the mgR mouse model of Marfan syndrome showed increased chemotactic stimulatory activity related to the elastin-binding protein. Aortic samples were collected from 6 patients with Marfan syndrome and 8 with isolated aneurysms of the ascending aorta. Control samples were obtained from 11 organ donors without known vascular or connective tissue diseases. Soluble proteins extracted from the aortic samples of the two patient groups were compared against buffer controls and against the aortic samples from controls with respect to the ability to induce macrophage chemotaxis as measured using a modified Boyden chamber, as well as the reactivity to a monoclonal antibody BA4 against bioactive elastin peptides using ELISA. Samples from Marfan patients displayed a statistically significant increase in chemotactic inductive activity compared to control samples. Additionally, reactivity to BA4 was significantly increased. Similar statistically significant increases were identified for the samples from patients with idiopathic thoracic aortic aneurysm. There was a significant correlation between the chemotactic index and BA4 reactivity, and the increases in chemotactic activity of extracts from Marfan patients could be inhibited by pretreatment with lactose, VGVAPG peptides, or BA4, which indicates the involvement of EBP in mediating the effects. Our results demonstrate that aortic extracts of patients with Marfan syndrome can elicit macrophage chemotaxis, similar to our previous study on aortic extracts of the mgR mouse model of Marfan syndrome (Guo et al., Circulation 2006; 114:1855-62). PMID- 21647418 TI - Multiple pH regime molecular dynamics simulation for pK calculations. AB - Ionisation equilibria in proteins are influenced by conformational flexibility, which can in principle be accounted for by molecular dynamics simulation. One problem in this method is the bias arising from the fixed protonation state during the simulation. Its effect is mostly exhibited when the ionisation behaviour of the titratable groups is extrapolated to pH regions where the predetermined protonation state of the protein may not be statistically relevant, leading to conformational sampling that is not representative of the true state. In this work we consider a simple approach which can essentially reduce this problem. Three molecular dynamics structure sets are generated, each with a different protonation state of the protein molecule expected to be relevant at three pH regions, and pK calculations from the three sets are combined to predict pK over the entire pH range of interest. This multiple pH molecular dynamics approach was tested on the GCN4 leucine zipper, a protein for which a full data set of experimental data is available. The pK values were predicted with a mean deviation from the experimental data of 0.29 pH units, and with a precision of 0.13 pH units, evaluated on the basis of equivalent sites in the dimeric GCN4 leucine zipper. PMID- 21647419 TI - Characterization of novel StAR (steroidogenic acute regulatory protein) mutations causing non-classic lipoid adrenal hyperplasia. AB - CONTEXT: Steroidogenic acute regulatory protein (StAR) is crucial for transport of cholesterol to mitochondria where biosynthesis of steroids is initiated. Loss of StAR function causes lipoid congenital adrenal hyperplasia (LCAH). OBJECTIVE: StAR gene mutations causing partial loss of function manifest atypical and may be mistaken as familial glucocorticoid deficiency. Only a few mutations have been reported. DESIGN: To report clinical, biochemical, genetic, protein structure and functional data on two novel StAR mutations, and to compare them with published literature. SETTING: Collaboration between the University Children's Hospital Bern, Switzerland, and the CIBERER, Hospital Vall d'Hebron, Autonomous University, Barcelona, Spain. PATIENTS: Two subjects of a non-consanguineous Caucasian family were studied. The 46,XX phenotypic normal female was diagnosed with adrenal insufficiency at the age of 10 months, had normal pubertal development and still has no signs of hypergonodatropic hypogonadism at 32 years of age. Her 46,XY brother was born with normal male external genitalia and was diagnosed with adrenal insufficiency at 14 months. Puberty was normal and no signs of hypergonadotropic hypogonadism are present at 29 years of age. RESULTS: StAR gene analysis revealed two novel compound heterozygote mutations T44HfsX3 and G221S. T44HfsX3 is a loss-of-function StAR mutation. G221S retains partial activity (~30%) and is therefore responsible for a milder, non-classic phenotype. G221S is located in the cholesterol binding pocket and seems to alter binding/release of cholesterol. CONCLUSIONS: StAR mutations located in the cholesterol binding pocket (V187M, R188C, R192C, G221D/S) seem to cause non classic lipoid CAH. Accuracy of genotype-phenotype prediction by in vitro testing may vary with the assays employed. PMID- 21647420 TI - Long-term continuous corticosterone treatment decreases VEGF receptor-2 expression in frontal cortex. AB - OBJECTIVE: Stress and increased glucocorticoid levels are associated with many neuropsychiatric disorders including schizophrenia and depression. Recently, the role of vascular endothelial factor receptor-2 (VEGFR2/Flk1) signaling has been implicated in stress-mediated neuroplasticity. However, the mechanism of regulation of VEGF/Flk1 signaling under long-term continuous glucocorticoid exposure has not been elucidated. MATERIAL AND METHODS: We examined the possible effects of long-term continuous glucocorticoid exposure on VEGF/Flk1 signaling in cultured cortical neurons in vitro, mouse frontal cortex in vivo, and in post mortem human prefrontal cortex of both control and schizophrenia subjects. RESULTS: We found that long-term continuous exposure to corticosterone (CORT, a natural glucocorticoid) reduced Flk1 protein levels both in vitro and in vivo. CORT treatment resulted in alterations in signaling molecules downstream to Flk1 such as PTEN, Akt and mTOR. We demonstrated that CORT-induced changes in Flk1 levels are mediated through glucocorticoid receptor (GR) and calcium. A significant reduction in Flk1-GR interaction was observed following CORT exposure. Interestingly, VEGF levels were increased in cortex, but decreased in serum following CORT treatment. Moreover, significant reductions in Flk1 and GR protein levels were found in postmortem prefrontal cortex samples from schizophrenia subjects. CONCLUSIONS: The alterations in VEGF/Flk1 signaling following long-term continuous CORT exposure represents a molecular mechanism of the neurobiological effects of chronic stress. PMID- 21647421 TI - IL-17RA is required for CCL2 expression, macrophage recruitment, and emphysema in response to cigarette smoke. AB - Chronic Obstructive Pulmonary Disease (COPD) is characterized by airspace enlargement and peribronchial lymphoid follicles; however, the immunological mechanisms leading to these pathologic changes remain undefined. Here we show that cigarette smoke is a selective adjuvant that augments in vitro and in vivo Th17, but not Th1, cell differentiation via the aryl hydrocarbon receptor. Smoke exposed IL-17RA(-/-) mice failed to induce CCL2 and MMP12 compared to WT mice. Remarkably, in contrast to WT mice, IL-17RA(-/-) mice failed to develop emphysema after 6 months of cigarette smoke exposure. Taken together, these data demonstrate that cigarette smoke is a potent Th17 adjuvant and that IL-17RA signaling is required for chemokine expression necessary for MMP12 induction and tissue emphysema. PMID- 21647422 TI - Biological surface coating and molting inhibition as mechanisms of TiO2 nanoparticle toxicity in Daphnia magna. AB - The production and use of nanoparticles (NP) has steadily increased within the last decade; however, knowledge about risks of NP to human health and ecosystems is still scarce. Common knowledge concerning NP effects on freshwater organisms is largely limited to standard short-term (<=48 h) toxicity tests, which lack both NP fate characterization and an understanding of the mechanisms underlying toxicity. Employing slightly longer exposure times (72 to 96 h), we found that suspensions of nanosized (~100 nm initial mean diameter) titanium dioxide (nTiO(2)) led to toxicity in Daphnia magna at nominal concentrations of 3.8 (72-h EC(50)) and 0.73 mg/L (96-h EC(50)). However, nTiO(2) disappeared quickly from the ISO-medium water phase, resulting in toxicity levels as low as 0.24 mg/L (96 h EC(50)) based on measured concentrations. Moreover, we showed that nTiO(2) (~100 nm) is significantly more toxic than non-nanosized TiO(2) (~200 nm) prepared from the same stock suspension. Most importantly, we hypothesized a mechanistic chain of events for nTiO(2) toxicity in D. magna that involves the coating of the organism surface with nTiO(2) combined with a molting disruption. Neonate D. magna (<=6 h) exposed to 2 mg/L nTiO(2) exhibited a "biological surface coating" that disappeared within 36 h, during which the first molting was successfully managed by 100% of the exposed organisms. Continued exposure up to 96 h led to a renewed formation of the surface coating and significantly reduced the molting rate to 10%, resulting in 90% mortality. Because coating of aquatic organisms by manmade NP might be ubiquitous in nature, this form of physical NP toxicity might result in widespread negative impacts on environmental health. PMID- 21647423 TI - Development of an orthotopic human pancreatic cancer xenograft model using ultrasound guided injection of cells. AB - Mice have been employed as models of cancer for over a century, providing significant advances in our understanding of this multifaceted family of diseases. In particular, orthotopic tumor xenograft mouse models are emerging as the preference for cancer research due to increased clinical relevance over subcutaneous mouse models. In the current study, we developed orthotopic pancreatic cancer xenograft models in mice by a minimally invasive method, ultrasound guided injection (USGI) comparable to highly invasive surgical orthotopic injection (SOI) methods. This optimized method prevented injection complications such as recoil of cells through the injection canal or leakage of cells out of the pancreas into the peritoneal cavity. Tumor growth was monitored in vivo and quantified by ultrasound imaging weekly, tumors were also detected by in vivo fluorescence imaging using a tumor targeted molecular probe. The mean tumor volumes for the USGI and SOI models after 2 weeks of tumor growth were 205 mm(3) and 178 mm(3) respectively. By USGI of human pancreatic cancer cell lines, human orthotopic pancreatic cancer xenografts were established. Based on ultrasound imaging, the orthotopic human pancreatic cancer xenograft take rate was 100% for both human pancreatic cancer cell lines used, MiaPaCa-2 and Su86.86, with mean tumor volumes of 28 mm(3)and 30 mm(3). We demonstrated that this USGI method is feasible, reproducible, facile, minimally invasive and improved compared to the highly-invasive SOI method for establishing orthotopic pancreatic tumor xenograft models suitable for molecular imaging. PMID- 21647424 TI - Insulin-like peptides and the target of rapamycin pathway coordinately regulate blood digestion and egg maturation in the mosquito Aedes aegypti. AB - BACKGROUND: Mosquitoes are insects that vector many serious pathogens to humans and other vertebrates. Most mosquitoes must feed on the blood of a vertebrate host to produce eggs. In turn, multiple cycles of blood feeding promote frequent contacts with hosts and make mosquitoes ideal disease vectors. Both hormonal and nutritional factors are involved in regulating egg development in the mosquito, Aedes aegypti. However, the processes that regulate digestion of the blood meal remain unclear. METHODOLOGY/PRINCIPAL FINDINGS: Here we report that insulin peptide 3 (ILP3) directly stimulated late phase trypsin-like gene expression in blood fed females. In vivo knockdown of the mosquito insulin receptor (MIR) by RNA interference (RNAi) delayed but did not fully inhibit trypsin-like gene expression in the midgut, ecdysteroid (ECD) production by ovaries, and vitellogenin (Vg) expression by the fat body. In contrast, in vivo treatment with double-stranded MIR RNA and rapamycin completely blocked egg production. In vitro experiments showed that amino acids did not simulate late phase trypsin-like gene expression in the midgut or ECD production by the ovaries. However, amino acids did enhance ILP3-mediated stimulation of trypsin-like gene expression and ECD production. CONCLUSIONS/SIGNIFICANCE: Overall, our results indicate that ILPs from the brain synchronize blood meal digestion and amino acid availability with ovarian ECD production to maximize Vg expression by the fat body. The activation of digestion by ILPs may also underlie the growth promoting effects of insulin and TOR signaling in other species. PMID- 21647426 TI - A propaganda index for reviewing problem framing in articles and manuscripts: an exploratory study. AB - OBJECTIVE: To determine the effectiveness of an index in increasing recognition of misleading problem framing in articles and manuscripts. DESIGN: A propaganda index consisting of 32 items was developed drawing on related literature. Seventeen subjects who review manuscripts for possible publication were requested to read five recent published reports of randomized controlled trials concerning social anxiety and to identify indicators of propaganda (defined as encouraging beliefs and actions with the least thought possible). They then re-read the same five articles using a propaganda index to note instances of propaganda. DATA SOURCE: Convenience sample of individuals who review manuscripts for possible publication and sample of recent published reports of randomized controlled trials regarding social anxiety in five different journals by different authors, blinded by author and journal. RESULTS: Data showed that there was a high rate of propagandistic problem framing in reports of RCTs regarding social anxiety such as hiding well argued alternative views and vagueness. This occurred in 117 out of 160 opportunities over five research reports. A convenience sample of 17 academics spotted only 4.5 percent of propaganda indicators. This increased to 64 percent with use of the 32 item propaganda index. Use of a propaganda index increased recognition of related indicators. However many instances remained undetected. CONCLUSION: This propaganda index warrants further exploration as a complement to reporting guidelines such as CONSORT and PRISMA. PMID- 21647425 TI - An international randomised placebo-controlled trial of a four-component combination pill ("polypill") in people with raised cardiovascular risk. AB - BACKGROUND: There has been widespread interest in the potential of combination cardiovascular medications containing aspirin and agents to lower blood pressure and cholesterol ('polypills') to reduce cardiovascular disease. However, no reliable placebo-controlled data are available on both efficacy and tolerability. METHODS: We conducted a randomised, double-blind placebo-controlled trial of a polypill (containing aspirin 75 mg, lisinopril 10 mg, hydrochlorothiazide 12.5 mg and simvastatin 20 mg) in 378 individuals without an indication for any component of the polypill, but who had an estimated 5-year cardiovascular disease risk over 7.5%. The primary outcomes were systolic blood pressure (SBP), LDL-cholesterol and tolerability (proportion discontinued randomised therapy) at 12 weeks follow up. FINDINGS: At baseline, mean BP was 134/81 mmHg and mean LDL-cholesterol was 3.7 mmol/L. Over 12 weeks, polypill treatment reduced SBP by 9.9 (95% CI: 7.7 to 12.1) mmHg and LDL-cholesterol by 0.8 (95% CI 0.6 to 0.9) mmol/L. The discontinuation rates in the polypill group compared to placebo were 23% vs 18% (RR 1.33, 95% CI 0.89 to 2.00, p = 0.2). There was an excess of side effects known to the component medicines (58% vs 42%, p = 0.001), which was mostly apparent within a few weeks, and usually did not warrant cessation of trial treatment. CONCLUSIONS: This polypill achieved sizeable reductions in SBP and LDL cholesterol but caused side effects in about 1 in 6 people. The halving in predicted cardiovascular risk is moderately lower than previous estimates and the side effect rate is moderately higher. Nonetheless, substantial net benefits would be expected among patients at high risk. TRIAL REGISTRATION: Australian New Zealand Clinical Trials Registry ACTRN12607000099426. PMID- 21647427 TI - Trends over 5 decades in U.S. occupation-related physical activity and their associations with obesity. AB - BACKGROUND: The true causes of the obesity epidemic are not well understood and there are few longitudinal population-based data published examining this issue. The objective of this analysis was to examine trends in occupational physical activity during the past 5 decades and explore how these trends relate to concurrent changes in body weight in the U.S. METHODOLOGY/PRINCIPAL FINDINGS: Analysis of energy expenditure for occupations in U.S. private industry since 1960 using data from the U.S. Bureau of Labor Statistics. Mean body weight was derived from the U.S. National Health and Nutrition Examination Surveys (NHANES). In the early 1960's almost half the jobs in private industry in the U.S. required at least moderate intensity physical activity whereas now less than 20% demand this level of energy expenditure. Since 1960 the estimated mean daily energy expenditure due to work related physical activity has dropped by more than 100 calories in both women and men. Energy balance model predicted weights based on change in occupation-related daily energy expenditure since 1960 for each NHANES examination period closely matched the actual change in weight for 40-50 year old men and women. For example from 1960-62 to 2003-06 we estimated that the occupation-related daily energy expenditure decreased by 142 calories in men. Given a baseline weight of 76.9 kg in 1960-02, we estimated that a 142 calories reduction would result in an increase in mean weight to 89.7 kg, which closely matched the mean NHANES weight of 91.8 kg in 2003-06. The results were similar for women. CONCLUSION: Over the last 50 years in the U.S. we estimate that daily occupation-related energy expenditure has decreased by more than 100 calories, and this reduction in energy expenditure accounts for a significant portion of the increase in mean U.S. body weights for women and men. PMID- 21647428 TI - Bioassays to monitor Taspase1 function for the identification of pharmacogenetic inhibitors. AB - BACKGROUND: Threonine Aspartase 1 (Taspase1) mediates cleavage of the mixed lineage leukemia (MLL) protein and leukemia provoking MLL-fusions. In contrast to other proteases, the understanding of Taspase1's (patho)biological relevance and function is limited, since neither small molecule inhibitors nor cell based functional assays for Taspase1 are currently available. METHODOLOGY/FINDINGS: Efficient cell-based assays to probe Taspase1 function in vivo are presented here. These are composed of glutathione S-transferase, autofluorescent protein variants, Taspase1 cleavage sites and rational combinations of nuclear import and export signals. The biosensors localize predominantly to the cytoplasm, whereas expression of biologically active Taspase1 but not of inactive Taspase1 mutants or of the protease Caspase3 triggers their proteolytic cleavage and nuclear accumulation. Compared to in vitro assays using recombinant components the in vivo assay was highly efficient. Employing an optimized nuclear translocation algorithm, the triple-color assay could be adapted to a high-throughput microscopy platform (Z'factor = 0.63). Automated high-content data analysis was used to screen a focused compound library, selected by an in silico pharmacophor screening approach, as well as a collection of fungal extracts. Screening identified two compounds, N-[2-[(4-amino-6-oxo-3H-pyrimidin-2 yl)sulfanyl]ethyl]benzenesulfonamide and 2-benzyltriazole-4,5-dicarboxylic acid, which partially inhibited Taspase1 cleavage in living cells. Additionally, the assay was exploited to probe endogenous Taspase1 in solid tumor cell models and to identify an improved consensus sequence for efficient Taspase1 cleavage. This allowed the in silico identification of novel putative Taspase1 targets. Those include the FERM Domain-Containing Protein 4B, the Tyrosine-Protein Phosphatase Zeta, and DNA Polymerase Zeta. Cleavage site recognition and proteolytic processing of these substrates were verified in the context of the biosensor. CONCLUSIONS: The assay not only allows to genetically probe Taspase1 structure function in vivo, but is also applicable for high-content screening to identify Taspase1 inhibitors. Such tools will provide novel insights into Taspase1's function and its potential therapeutic relevance. PMID- 21647429 TI - Behavior, color change and time for sexual inversion in the protogynous grouper (Epinephelus adscensionis). AB - Hermaphroditism, associated with territoriality and dominance behavior, is common in the marine environment. While male sex-specific coloration patterns have been documented in groupers, particularly during the spawning season, few data regarding social structure and the context for these color displays are available. In the present study, we define the social structure and male typical behavior of rock hind (Epinephelus adscensionis) in the wild. In addition, we detail the captive conditions and time period necessary to induce the onset of the sex-specific coloration and sexual change. At six oil production platform locations in the Gulf of Mexico, rock hind social group size and typical male rock hind social behavior were documented. We observed a rapid temporary color display in rock hind that could be turned on and off within three seconds and was used for confronting territory intruders and displays of aggression towards females. The male-specific "tuxedo" pattern consists of a bright yellow tail, a body with alternating dark brown and white patches and a dark bar extending from the upper mandible to the operculum. Identification and size ranges of male, female and intersex fish collected from oil platforms were determined in conjunction with gonadal histology. Rock hind social order is haremic with one dominant male defending a territory and a linear dominance hierarchy among individuals. In five captive experiments, the largest remaining female rock hind displayed the male specific color pattern within 32d after dominant male removal from the social group. To our knowledge, this is the first evidence in a grouper species of color patterning used to display territoriality and dominance outside of spawning aggregations. The behavioral paradigm described here is a key advance that will enable mechanistic studies of this complex sex change process. PMID- 21647430 TI - Eyes are windows to the Chinese soul: evidence from the detection of real and fake smiles. AB - How do people interpret the meaning of a smile? Previous studies with Westerners have found that both the eyes and the mouth are crucial in identifying and interpreting smiles, yet less is known about Easterners. Here we reported that when asking the Chinese to judge the Duchenne and non-Duchenne smiles as either real or fake, their accuracy and sensitivity were negatively correlated with their individualism scores but positively correlated with their collectivism scores. However, such correlations were found only for participants who stated the eyes to be the most useful references, but not for those who favored the mouth. Moreover, participants who favored the eyes were more accurate and sensitive than those who favored the mouth. Our results thus indicate that Chinese who follow the typical Eastern decoding process of using the eyes as diagnostic cues to identify and interpret others' facial expressions and social intentions, are particularly accurate and sensitive, the more they self-report greater collectivistic and lower individualistic values. PMID- 21647431 TI - Skin barrier homeostasis in atopic dermatitis: feedback regulation of kallikrein activity. AB - Atopic dermatitis (AD) is a widely spread cutaneous chronic disease characterised by sensitive reactions (eg. eczema) to normally innocuous elements. Although relatively little is understood about its underlying mechanisms due to its complexity, skin barrier dysfunction has been recognised as a key factor in the development of AD. Skin barrier homeostasis requires tight control of the activity of proteases, called kallikreins (KLKs), whose activity is regulated by a complex network of protein interactions that remains poorly understood despite its pathological importance. Characteristic symptoms of AD include the outbreak of inflammation triggered by external (eg. mechanical and chemical) stimulus and the persistence and aggravation of inflammation even if the initial stimulus disappears. These characteristic symptoms, together with some experimental data, suggest the presence of positive feedback regulation for KLK activity by inflammatory signals. We developed simple mathematical models for the KLK activation system to study the effects of feedback loops and carried out bifurcation analysis to investigate the model behaviours corresponding to inflammation caused by external stimulus. The model analysis confirmed that the hypothesised core model mechanisms capture the essence of inflammation outbreak by a defective skin barrier. Our models predicted the outbreaks of inflammation at weaker stimulus and its longer persistence in AD patients compared to healthy control. We also proposed a novel quantitative indicator for inflammation level by applying principal component analysis to microarray data. The model analysis reproduced qualitative AD characteristics revealed by this indicator. Our results strongly implicate the presence and importance of feedback mechanisms in KLK activity regulation. We further proposed future experiments that may provide informative data to enhance the system-level understanding on the regulatory mechanisms of skin barrier in AD and healthy individuals. PMID- 21647432 TI - Analysing dynamical behavior of cellular networks via stochastic bifurcations. AB - The dynamical structure of genetic networks determines the occurrence of various biological mechanisms, such as cellular differentiation. However, the question of how cellular diversity evolves in relation to the inherent stochasticity and intercellular communication remains still to be understood. Here, we define a concept of stochastic bifurcations suitable to investigate the dynamical structure of genetic networks, and show that under stochastic influence, the expression of given proteins of interest is defined via the probability distribution of the phase variable, representing one of the genes constituting the system. Moreover, we show that under changing stochastic conditions, the probabilities of expressing certain concentration values are different, leading to different functionality of the cells, and thus to differentiation of the cells in the various types. PMID- 21647433 TI - Trees wanted--dead or alive! Host selection and population dynamics in tree killing bark beetles. AB - Bark beetles (Coleoptera: Curculionidae, Scolytinae) feed and breed in dead or severely weakened host trees. When their population densities are high, some species aggregate on healthy host trees so that their defences may be exhausted and the inner bark successfully colonized, killing the tree in the process. Here we investigate under what conditions participating with unrelated conspecifics in risky mass attacks on living trees is an adaptive strategy, and what this can tell us about bark beetle outbreak dynamics. We find that the outcome of individual host selection may deviate from the ideal free distribution in a way that facilitates the emergence of tree-killing (aggressive) behavior, and that any heritability on traits governing aggressiveness seems likely to exist in a state of flux or cycles consistent with variability observed in natural populations. This may have implications for how economically and ecologically important species respond to environmental changes in climate and landscape (forest) structure. The population dynamics emerging from individual behavior are complex, capable of switching between "endemic" and "epidemic" regimes spontaneously or following changes in host availability or resistance. Model predictions are compared to empirical observations, and we identify some factors determining the occurrence and self-limitation of epidemics. PMID- 21647434 TI - Role of mitochondrial electron transport chain complexes in capsaicin mediated oxidative stress leading to apoptosis in pancreatic cancer cells. AB - We evaluated the mechanism of capsaicin-mediated ROS generation in pancreatic cancer cells. The generation of ROS was about 4-6 fold more as compared to control and as early as 1 h after capsaicin treatment in BxPC-3 and AsPC-1 cells but not in normal HPDE-6 cells. The generation of ROS was inhibited by catalase and EUK-134. To delineate the mechanism of ROS generation, enzymatic activities of mitochondrial complex-I and complex-III were determined in the pure mitochondria. Our results shows that capsaicin inhibits about 2.5-9% and 5-20% of complex-I activity and 8-75% of complex-III activity in BxPC-3 and AsPC-1 cells respectively, which was attenuable by SOD, catalase and EUK-134. On the other hand, capsaicin treatment failed to inhibit complex-I or complex-III activities in normal HPDE-6 cells. The ATP levels were drastically suppressed by capsaicin treatment in both BxPC-3 and AsPC-1 cells and attenuated by catalase or EUK-134. Oxidation of mitochondria-specific cardiolipin was substantially higher in capsaicin treated cells. BxPC-3 derived rho(0) cells, which lack mitochondrial DNA, were completely resistant to capsaicin mediated ROS generation and apoptosis. Our results reveal that the release of cytochrome c and cleavage of both caspase-9 and caspase-3 due to disruption of mitochondrial membrane potential were significantly blocked by catalase and EUK-134 in BxPC-3 cells. Our results further demonstrate that capsaicin treatment not only inhibit the enzymatic activity and expression of SOD, catalase and glutathione peroxidase but also reduce glutathione level. Over-expression of catalase by transient transfection protected the cells from capsaicin-mediated ROS generation and apoptosis. Furthermore, tumors from mice orally fed with 2.5 mg/kg capsaicin show decreased SOD activity and an increase in GSSG/GSH levels as compared to controls. Taken together, our results suggest the involvement of mitochondrial complex-I and III in capsaicin-mediated ROS generation and decrease in antioxidant levels resulting in severe mitochondrial damage leading to apoptosis in pancreatic cancer cells. PMID- 21647435 TI - Melatonin induces follicle maturation in Danio rerio. AB - Most organisms modulate their reproductive activity responding to day length by the nocturnal release of melatonin by the pineal gland. This hormone is also responsible for synchronizing reproduction with specific external environment stimuli in order to optimize reproductive success.The aim of this study was to establish the effect of melatonin on zebrafish reproduction.Adult females were daily exposed, via water, to two different doses (100 nM and 1 uM) of melatonin. Melatonin led to an increase of the Gonado Somatic Index (GSI) associated with the increase of eggs production, and the raise of gene and protein levels of vitellogenin (VTG) and estradiol receptor alpha (ERalpha) in the liver. The ability of melatonin to increase fecundity was consistent with a significant increase of gene transcription of kiss 1, kiss 2, gnrh3, in the brain, and lh in the pituitary, while in the ovary (in class IIIB follicles), with a significant decrease of two genes codifying for intra-ovarian regulators of premature oocyte maturation, the tgfbeta1 and the bmp15. The reduction in the expression of these two genes was concomitant with the increase of lhr and a modulation of mpralpha and mprbeta gene transcription, whose proteins are involved in oocyte maturation. Melatonin also exerted a direct action on follicles as shown by the increase of the oocytes undergoing to germinal vesicle break down (GVBD) and modulated mpr alpha and beta gene expression in the in vitro exposure.These data highlight the effects of melatonin in promoting zebrafish reproduction exerting its effects either in the brain-pituitary and in the gonads. PMID- 21647436 TI - The hexameric structures of human heat shock protein 90. AB - BACKGROUND: The human 90-kDa heat shock protein (HSP90) functions as a dimeric molecular chaperone. HSP90 identified on the cell surface has been found to play a crucial role in cancer invasion and metastasis, and has become a validated anti cancer target for drug development. It has been shown to self-assemble into oligomers upon heat shock or divalent cations treatment, but the functional role of the oligomeric states in the chaperone cycle is not fully understood. PRINCIPAL FINDINGS: Here we report the crystal structure of a truncated HSP90 that contains the middle segment and the carboxy-terminal domain, termed MC HSP90. The structure reveals an architecture with triangular bipyramid geometry, in which the building block of the hexameric assembly is a dimer. In solution, MC HSP90 exists in three major oligomer states, namely dimer, tetramer and hexamer, which were elucidated by size exclusion chromatography and analytical ultracentrifugation. The newly discovered HSP90 isoform HSP90N that lacks the N terminal ATPase domain also exhibited similar oligomerization states as did MC HSP90. CONCLUSIONS: While lacking the ATPase domain, both MC-HSP90 and HSP90N can self-assemble into a hexameric structure, spontaneously. The crystal structure of MC-HSP90 reveals that, in addition to the C-terminal dimerization domain, the residue W320 in the M domain plays a critical role in its oligomerization. This study not only demonstrates how the human MC-HSP90 forms a hexamer, but also justifies the similar formation of HSP90N by using 3D modeling analysis. PMID- 21647437 TI - Television viewing and incident cardiovascular disease: prospective associations and mediation analysis in the EPIC Norfolk Study. AB - BACKGROUND: Although television viewing time is detrimentally associated with intermediate cardiovascular risk factors, the relationship with incident total (i.e. combined fatal and non-fatal) cardiovascular disease (CVD), non-fatal CVD and coronary heart disease is largely unknown. This study examined whether television viewing time is associated with these three outcomes, independently of physical activity energy expenditure and other confounding variables. METHODOLOGY/PRINCIPAL FINDINGS: A population-based cohort of 12,608 men and women (aged 61.4+/-9.0), free from stroke, myocardial infarction and cancer at baseline in 1998-2000 were followed up until 2007 (6.9+/-1.9 years). Participants self reported education, smoking, alcohol use, antihypertensive, lipid lowering and antidepressant medication, disease history, total energy intake, sleep duration, physical activity and television viewing. BMI, waist circumference, blood pressure, triglycerides, HDL cholesterol and glycated haemoglobin (HbA(1c)) were measured by standardized procedures; a clustered metabolic risk score was constructed. Every one hour/day increase in television viewing was associated with an increased hazard for total (HR = 1.06, 95%CI = 1.03-1.08; 2,620 cases), non-fatal CVD (HR = 1.06, 95%CI = 1.03-1.09; 2,134 cases), and coronary heart disease (HR = 1.08, 95%CI = 1.03-1.13; 940 cases), independent of gender, age, education, smoking, alcohol, medication, diabetes status, CVD family history, sleep duration and physical activity energy expenditure. Energy intake, BMI, waist circumference, blood pressure, triglycerides, HDL cholesterol, HbA(1c) and the clustered metabolic risk score only partially mediated these associations. CONCLUSIONS: These results indicate that the most prevalent leisure time (sedentary) behaviour, television viewing, independently contributes to increased CVD risk. Recommendations on reducing television viewing time should be considered. PMID- 21647438 TI - Nitric oxide synthase inhibition enhances the antitumor effect of radiation in the treatment of squamous carcinoma xenografts. AB - This study tests whether the nitric oxide synthase (NOS) inhibitor, N(G)-nitro-L arginine (L-NNA), combines favorably with ionizing radiation (IR) in controlling squamous carcinoma tumor growth. Animals bearing FaDu and A431 xenografts were treated with L-NNA in the drinking water. IR exposure was 10 Gy for tumor growth and survival studies and 4 Gy for ex vivo clonogenic assays. Cryosections were examined immunohistochemically for markers of apoptosis and hypoxia. Blood flow was assayed by fluorescent microscopy of tissue cryosections after i.v. injection of fluorospheres. Orally administered L-NNA for 24 hrs reduces tumor blood flow by 80% (p<0.01). Within 24 hrs L-NNA treatment stopped tumor growth for at least 10 days before tumor growth again ensued. The growth arrest was in part due to increased cell killing since a combination of L-NNA and a single 4 Gy IR caused 82% tumor cell killing measured by an ex vivo clonogenic assay compared to 49% by L-NNA or 29% by IR alone. A Kaplan-Meyer analysis of animal survival revealed a distinct survival advantage for the combined treatment. Combining L-NNA and IR was also found to be at least as effective as a single i.p. dose of cisplatin plus IR. In contrast to the in vivo studies, exposure of cells to L-NNA in vitro was without effect on clonogenicity with or without IR. Western and immunochemical analysis of expression of a number of proteins involved in NO signaling indicated that L-NNA treatment enhanced arginase-2 expression and that this may represent vasculature remodeling and escape from NOS inhibition. For tumors such as head and neck squamous carcinomas that show only modest responses to inhibitors of specific angiogenic pathways, targeting NO-dependent pro survival and angiogenic mechanisms in both tumor and supporting stromal cells may present a potential new strategy for tumor control. PMID- 21647439 TI - AP-1 is a component of the transcriptional network regulated by GSK-3 in quiescent cells. AB - BACKGROUND: The protein kinase GSK-3 is constitutively active in quiescent cells in the absence of growth factor signaling. Previously, we identified a set of genes that required GSK-3 to maintain their repression during quiescence. Computational analysis of the upstream sequences of these genes predicted transcription factor binding sites for CREB, NFkappaB and AP-1. In our previous work, contributions of CREB and NFkappaB were examined. In the current study, the AP-1 component of the signaling network in quiescent cells was explored. METHODOLOGY/PRINCIPAL FINDINGS: Using chromatin immunoprecipitation analysis, two AP-1 family members, c-Jun and JunD, bound to predicted upstream regulatory sequences in 8 of the 12 GSK-3-regulated genes. c-Jun was phosphorylated on threonine 239 by GSK-3 in quiescent cells, consistent with previous studies demonstrating inhibition of c-Jun by GSK-3. Inhibition of GSK-3 attenuated this phosphorylation, resulting in the stabilization of c-Jun. The association of c Jun with its target sequences was increased by growth factor stimulation as well as by direct GSK-3 inhibition. The physiological role for c-Jun was also confirmed by siRNA inhibition of gene induction. CONCLUSIONS/SIGNIFICANCE: These results indicate that inhibition of c-Jun by GSK-3 contributes to the repression of growth factor-inducible genes in quiescent cells. Together, AP-1, CREB and NFkappaB form an integrated transcriptional network that is largely responsible for maintaining repression of target genes downstream of GSK-3 signaling. PMID- 21647440 TI - Charge isomers of myelin basic protein: structure and interactions with membranes, nucleotide analogues, and calmodulin. AB - As an essential structural protein required for tight compaction of the central nervous system myelin sheath, myelin basic protein (MBP) is one of the candidate autoantigens of the human inflammatory demyelinating disease multiple sclerosis, which is characterized by the active degradation of the myelin sheath. In this work, recombinant murine analogues of the natural C1 and C8 charge components (rmC1 and rmC8), two isoforms of the classic 18.5-kDa MBP, were used as model proteins to get insights into the structure and function of the charge isomers. Various biochemical and biophysical methods such as size exclusion chromatography, calorimetry, surface plasmon resonance, small angle X-ray and neutron scattering, Raman and fluorescence spectroscopy, and conventional as well as synchrotron radiation circular dichroism were used to investigate differences between these two isoforms, both from the structural point of view, and regarding interactions with ligands, including calmodulin (CaM), various detergents, nucleotide analogues, and lipids. Overall, our results provide further proof that rmC8 is deficient both in structure and especially in function, when compared to rmC1. While the CaM binding properties of the two forms are very similar, their interactions with membrane mimics are different. CaM can be used to remove MBP from immobilized lipid monolayers made of synthetic lipids--a phenomenon, which may be of relevance for MBP function and its regulation. Furthermore, using fluorescently labelled nucleotides, we observed binding of ATP and GTP, but not AMP, by MBP; the binding of nucleoside triphosphates was inhibited by the presence of CaM. Together, our results provide important further data on the interactions between MBP and its ligands, and on the differences in the structure and function between MBP charge isomers. PMID- 21647441 TI - Avian ultraviolet/violet cones identified as probable magnetoreceptors. AB - BACKGROUND: The Radical-Pair-Model postulates that the reception of magnetic compass directions in birds is based on spin-chemical reactions in specialized photopigments in the eye, with cryptochromes discussed as candidate molecules. But so far, the exact subcellular characterization of these molecules in the retina remained unknown. METHODOLOGY/PRINCIPAL FINDINGS: We here describe the localization of cryptochrome 1a (Cry1a) in the retina of European robins, Erithacus rubecula, and domestic chickens, Gallus gallus, two species that have been shown to use the magnetic field for compass orientation. In both species, Cry1a is present exclusively in the ultraviolet/violet (UV/V) cones that are distributed across the entire retina. Electron microscopy shows Cry1a in ordered bands along the membrane discs of the outer segment, and cell fractionation reveals Cry1a in the membrane fraction, suggesting the possibility that Cry1a is anchored along membranes. CONCLUSIONS/SIGNIFICANCE: We provide first structural evidence that Cry1a occurs within a sensory structure arranged in a way that fulfils essential requirements of the Radical-Pair-Model. Our findings, identifying the UV/V-cones as probable magnetoreceptors, support the assumption that Cry1a is indeed the receptor molecule mediating information on magnetic directions, and thus provide the Radical-Pair-Model with a profound histological background. PMID- 21647442 TI - Polymorphisms of homologous recombination genes and clinical outcomes of non small cell lung cancer patients treated with definitive radiotherapy. AB - The repair of DNA double-strand breaks (DSBs) is the major mechanism to maintain genomic stability in response to irradiation. We hypothesized that genetic polymorphisms in DSB repair genes may affect clinical outcomes among non-small cell lung cancer (NSCLC) patients treated with definitive radio(chemo)therapy. We genotyped six potentially functional single nucleotide polymorphisms (SNPs) (i.e., RAD51 -135G>C/rs1801320 and -172G>T/rs1801321, XRCC2 4234G>C/rs3218384 and R188H/rs3218536 G>A, XRCC3 T241M/rs861539 and NBN E185Q/rs1805794) and estimated their associations with overall survival (OS) and radiation pneumonitis (RP) in 228 NSCLC patients. We found a predictive role of RAD51 -135G>C SNP in RP development (adjusted hazard ratio [HR] = 0.52, 95% confidence interval [CI], 0.31-0.86, P = 0.010 for CG/CC vs. GG). We also found that RAD51 -135G>C and XRCC2 R188H SNPs were independent prognostic factors for overall survival (adjusted HR = 1.70, 95% CI, 1.14-2.62, P = 0.009 for CG/CC vs. GG; and adjusted HR = 1.70; 95% CI, 1.02-2.85, P = 0.043 for AG vs. GG, respectively) and that the SNP-survival association was most pronounced in the presence of RP. Our study suggests that HR genetic polymorphisms, particularly RAD51 -135G>C, may influence overall survival and radiation pneumonitis in NSCLC patients treated with definitive radio(chemo)therapy. Large studies are needed to confirm our findings. PMID- 21647443 TI - A hierarchical probabilistic model for rapid object categorization in natural scenes. AB - Humans can categorize objects in complex natural scenes within 100-150 ms. This amazing ability of rapid categorization has motivated many computational models. Most of these models require extensive training to obtain a decision boundary in a very high dimensional (e.g., ~6,000 in a leading model) feature space and often categorize objects in natural scenes by categorizing the context that co-occurs with objects when objects do not occupy large portions of the scenes. It is thus unclear how humans achieve rapid scene categorization.To address this issue, we developed a hierarchical probabilistic model for rapid object categorization in natural scenes. In this model, a natural object category is represented by a coarse hierarchical probability distribution (PD), which includes PDs of object geometry and spatial configuration of object parts. Object parts are encoded by PDs of a set of natural object structures, each of which is a concatenation of local object features. Rapid categorization is performed as statistical inference. Since the model uses a very small number (~100) of structures for even complex object categories such as animals and cars, it requires little training and is robust in the presence of large variations within object categories and in their occurrences in natural scenes. Remarkably, we found that the model categorized animals in natural scenes and cars in street scenes with a near human level performance. We also found that the model located animals and cars in natural scenes, thus overcoming a flaw in many other models which is to categorize objects in natural context by categorizing contextual features. These results suggest that coarse PDs of object categories based on natural object structures and statistical operations on these PDs may underlie the human ability to rapidly categorize scenes. PMID- 21647444 TI - Patterns of GPS tracks suggest nocturnal foraging by incubating Peruvian pelicans (Pelecanus thagus). AB - Most seabirds are diurnal foragers, but some species may also feed at night. In Peruvian pelicans (Pelecanus thagus), the evidence for nocturnal foraging is sparse and anecdotal. We used GPS-dataloggers on five incubating Peruvian pelicans from Isla Lobos de Tierra, Peru, to examine their nocturnality, foraging movements and activities patterns at sea. All instrumented pelicans undertook nocturnal trips during a 5-7 day tracking period. Eighty-seven percent of these trips (n = 13) were strictly nocturnal, whereas the remaining occurred during the day and night. Most birds departed from the island after sunset and returned a few hours after sunrise. Birds traveled south of the island for single-day trips at a maximum range of 82.8 km. Overall, 22% of the tracking period was spent at sea, whereas the remaining time was spent on the island. In the intermediate section of the trip (between inbound and outbound commutes), birds spent 77% of the trip time in floating bouts interspersed by short flying bouts, the former being on average three times longer than the latter. Taken together, the high sinuosity of the bird's tracks during floating bouts, the exclusively nocturnal trips of most individuals, and the fact that all birds returned to the island within a few hours after sunrise suggest that pelicans were actively feeding at night. The nocturnal foraging strategy of Peruvian pelicans may reduce food competition with the sympatric and strictly diurnal Guanay cormorants (Phalacrocorax bougainvillii), Peruvian boobies (Sula variegata) and Blue-footed boobies (S. nebouxii), which were present on the island in large numbers. Likewise, plankton bioluminescence might be used by pelicans as indirect cues to locate anchovies during their upward migration at night. The foraging success of pelicans at night may be enhanced by seizing prey close to the sea surface using a sit-and-wait strategy. PMID- 21647445 TI - HIVToolbox, an integrated web application for investigating HIV. AB - Many bioinformatic databases and applications focus on a limited domain of knowledge federating links to information in other databases. This segregated data structure likely limits our ability to investigate and understand complex biological systems. To facilitate research, therefore, we have built HIVToolbox, which integrates much of the knowledge about HIV proteins and allows virologists and structural biologists to access sequence, structure, and functional relationships in an intuitive web application. HIV-1 integrase protein was used as a case study to show the utility of this application. We show how data integration facilitates identification of new questions and hypotheses much more rapid and convenient than current approaches using isolated repositories. Several new hypotheses for integrase were created as an example, and we experimentally confirmed a predicted CK2 phosphorylation site. Weblink: [http://hivtoolbox.bio toolkit.com]. PMID- 21647446 TI - Simultaneous visualization of both signaling cascade activity and end-point gene expression in single cells. AB - We have developed an approach for simultaneous detection of individual endogenous protein modifications and mRNA molecules in single cells in situ. For this purpose we combined two methods previously developed in our lab: in situ proximity ligation assay for the detection of individual protein interactions and -modifications and in situ detection of single mRNA molecules using padlock probes. As proof-of-principle, we demonstrated the utility of the method for simultaneous detection of phosphorylated PDGFRbeta and DUSP6/MKP-3 mRNA molecules in individual human fibroblasts upon PDGF-BB stimulation. Further we applied drugs disrupting the PDGFRbeta signaling pathway at various sites to show that this combined method can concurrently monitor the molecular effect of the drugs, i.e. inhibition of downstream signaling from the targeted node in the signaling pathway. Due to its ability to detect different types of molecules in single cells in situ the method presented here can contribute to a deeper understanding of cell-to-cell variations and can be applied to e.g. pinpoint effector sites of drugs in a signaling pathway. PMID- 21647447 TI - PCR master mixes harbour murine DNA sequences. Caveat emptor! AB - BACKGROUND: XMRV is the most recently described retrovirus to be found in Man, firstly in patients with prostate cancer (PC) and secondly in 67% of patients with chronic fatigue syndrome (CFS) and 3.7% of controls. Both disease associations remain contentious. Indeed, a recent publication has concluded that "XMRV is unlikely to be a human pathogen". Subsequently related but different polytropic MLV (pMLV) sequences were also reported from the blood of 86.5% of patients with CFS. and 6.8% of controls. Consequently we decided to investigate blood donors for evidence of XMRV/pMLV. METHODOLOGY/PRINCIPAL FINDINGS: Testing of cDNA prepared from the whole blood of 80 random blood donors, generated gag PCR signals from two samples (7C and 9C). These had previously tested negative for XMRV by two other PCR based techniques. To test whether the PCR mix was the source of these sequences 88 replicates of water were amplified using Invitrogen Platinum Taq (IPT) and Applied Biosystems Taq Gold LD (ABTG). Four gag sequences (2D, 3F, 7H, 12C) were generated with the IPT, a further sequence (12D) by ABTG re-amplification of an IPT first round product. Sequence comparisons revealed remarkable similarities between these sequences, endogeous MLVs and the pMLV sequences reported in patients with CFS. CONCLUSIONS/SIGNIFICANCE: Methodologies for the detection of viruses highly homologous to endogenous murine viruses require special caution as the very reagents used in the detection process can be a source of contamination and at a level where it is not immediately apparent. It is suggested that such contamination is likely to explain the apparent presence of pMLV in CFS. PMID- 21647448 TI - OrthoList: a compendium of C. elegans genes with human orthologs. AB - BACKGROUND: C. elegans is an important model for genetic studies relevant to human biology and disease. We sought to assess the orthology between C. elegans and human genes to understand better the relationship between their genomes and to generate a compelling list of candidates to streamline RNAi-based screens in this model. RESULTS: We performed a meta-analysis of results from four orthology prediction programs and generated a compendium, "OrthoList", containing 7,663 C. elegans protein-coding genes. Various assessments indicate that OrthoList has extensive coverage with low false-positive and false-negative rates. Part of this evaluation examined the conservation of components of the receptor tyrosine kinase, Notch, Wnt, TGF-beta and insulin signaling pathways, and led us to update compendia of conserved C. elegans kinases, nuclear hormone receptors, F-box proteins, and transcription factors. Comparison with two published genome-wide RNAi screens indicated that virtually all of the conserved hits would have been obtained had just the OrthoList set (~38% of the genome) been targeted. We compiled Ortholist by InterPro domains and Gene Ontology annotation, making it easy to identify C. elegans orthologs of human disease genes for potential functional analysis. CONCLUSIONS: We anticipate that OrthoList will be of considerable utility to C. elegans researchers for streamlining RNAi screens, by focusing on genes with apparent human orthologs, thus reducing screening effort by ~60%. Moreover, we find that OrthoList provides a useful basis for annotating orthology and reveals more C. elegans orthologs of human genes in various functional groups, such as transcription factors, than previously described. PMID- 21647449 TI - Distinct CD4+ CD8+ double-positive T cells in the blood and liver of patients during chronic hepatitis B and C. AB - CD4(+) and CD8(+) T cells, the main effectors of adaptive cellular immune responses, differentiate from immature, non-functional CD4(+)CD8(+) double positive T (DPT) cells in the thymus. Increased proportions of circulating DPT lymphocytes have been observed during acute viral infections; in chronic viral diseases, the role and repartition of extra-thymic DPT cells remain largely uncharacterized. We performed a phenotypic analysis of DPT cells in blood and liver from patients chronically infected by hepatitis C (HCV) or B (HBV) viruses. The highest percentages of DPT cells, predominantly CD4(high)CD8(low), were observed in patients infected by HCV, while HBV-infected patients mostly displayed CD4(low)CD8(high) and CD4(high)CD8(high) DPT cells. All proportions of DPT cells were higher in liver than in blood with, for each subpopulation referred to above, a correlation between their frequencies in these two compartments. In HCV patients, intra-hepatic DPT cells displayed more heterogeneous activation, differentiation and memory phenotypes than in the blood; most of them expressed CD1a, a marker of T cell development in the thymus. Ex vivo, the inoculation of liver slices with HCV produced in cell culture was accompanied by a disappearance of CD8(high) cells, suggesting a direct effect of the virus on the phenotype of DPT cells in the liver. Our results suggest that, in half of the patients, chronic HCV infection promotes the production of DPT cells, perhaps by their re-induction in the thymus and selection in the liver. PMID- 21647450 TI - Exponential random graph modeling for complex brain networks. AB - Exponential random graph models (ERGMs), also known as p* models, have been utilized extensively in the social science literature to study complex networks and how their global structure depends on underlying structural components. However, the literature on their use in biological networks (especially brain networks) has remained sparse. Descriptive models based on a specific feature of the graph (clustering coefficient, degree distribution, etc.) have dominated connectivity research in neuroscience. Corresponding generative models have been developed to reproduce one of these features. However, the complexity inherent in whole-brain network data necessitates the development and use of tools that allow the systematic exploration of several features simultaneously and how they interact to form the global network architecture. ERGMs provide a statistically principled approach to the assessment of how a set of interacting local brain network features gives rise to the global structure. We illustrate the utility of ERGMs for modeling, analyzing, and simulating complex whole-brain networks with network data from normal subjects. We also provide a foundation for the selection of important local features through the implementation and assessment of three selection approaches: a traditional p-value based backward selection approach, an information criterion approach (AIC), and a graphical goodness of fit (GOF) approach. The graphical GOF approach serves as the best method given the scientific interest in being able to capture and reproduce the structure of fitted brain networks. PMID- 21647451 TI - Discovery of protein phosphorylation motifs through exploratory data analysis. AB - BACKGROUND: The need for efficient algorithms to uncover biologically relevant phosphorylation motifs has become very important with rapid expansion of the proteomic sequence database along with a plethora of new information on phosphorylation sites. Here we present a novel unsupervised method, called Motif Finder (in short, F-Motif) for identification of phosphorylation motifs. F-Motif uses clustering of sequence information represented by numerical features that exploit the statistical information hidden in some foreground data. Furthermore, these identified motifs are then filtered to find "actual" motifs with statistically significant motif scores. RESULTS AND DISCUSSION: We have applied F Motif to several new and existing data sets and compared its performance with two well known state-of-the-art methods. In almost all cases F-Motif could identify all statistically significant motifs extracted by the state-of-the-art methods. More importantly, in addition to this, F-Motif uncovers several novel motifs. We have demonstrated using clues from the literature that most of these new motifs discovered by F-Motif are indeed novel. We have also found some interesting phenomena. For example, for CK2 kinase, the conserved sites appear only on the right side of S. However, for CDK kinase, the adjacent site on the right of S is conserved with residue P. In addition, three different encoding methods, including a novel position contrast matrix (PCM) and the simplest binary coding, are used and the ability of F-motif to discover motifs remains quite robust with respect to encoding schemes. CONCLUSIONS: An iterative algorithm proposed here uses exploratory data analysis to discover motifs from phosphorylated data. The effectiveness of F-Motif has been demonstrated using several real data sets as well as using a synthetic data set. The method is quite general in nature and can be used to find other types of motifs also. We have also provided a server for F Motif at http://f-motif.classcloud.org/, http://bio.classcloud.org/f-motif/ or http://ymu.classcloud.org/f-motif/. PMID- 21647452 TI - Identification of B cell epitopes of alcohol dehydrogenase allergen of Curvularia lunata. AB - BACKGROUND/OBJECTIVE: Epitope identification assists in developing molecules for clinical applications and is useful in defining molecular features of allergens for understanding structure/function relationship. The present study was aimed to identify the B cell epitopes of alcohol dehydrogenase (ADH) allergen from Curvularia lunata using in-silico methods and immunoassay. METHOD: B cell epitopes of ADH were predicted by sequence and structure based methods and protein-protein interaction tools while T cell epitopes by inhibitory concentration and binding score methods. The epitopes were superimposed on a three dimensional model of ADH generated by homology modeling and analyzed for antigenic characteristics. Peptides corresponding to predicted epitopes were synthesized and immunoreactivity assessed by ELISA using individual and pooled patients' sera. RESULT: The homology model showed GroES like catalytic domain joined to Rossmann superfamily domain by an alpha helix. Stereochemical quality was confirmed by Procheck which showed 90% residues in most favorable region of Ramachandran plot while Errat gave a quality score of 92.733%. Six B cell (P1-P6) and four T cell (P7-P10) epitopes were predicted by a combination of methods. Peptide P2 (epitope P2) showed E(X)(2)GGP(X)(3)KKI conserved pattern among allergens of pathogenesis related family. It was predicted as high affinity binder based on electronegativity and low hydrophobicity. The computational methods employed were validated using Bet v 1 and Der p 2 allergens where 67% and 60% of the epitope residues were predicted correctly. Among B cell epitopes, Peptide P2 showed maximum IgE binding with individual and pooled patients' sera (mean OD 0.604+/-0.059 and 0.506+/-0.0035, respectively) followed by P1, P4 and P3 epitopes. All T cell epitopes showed lower IgE binding. CONCLUSION: Four B cell epitopes of C. lunata ADH were identified. Peptide P2 can serve as a potential candidate for diagnosis of allergic diseases. PMID- 21647453 TI - Rtt107 phosphorylation promotes localisation to DNA double-stranded breaks (DSBs) and recombinational repair between sister chromatids. AB - Efficient repair of DNA double-stranded breaks (DSB) requires a coordinated response at the site of lesion. Nucleolytic resection commits repair towards homologous recombination, which preferentially occurs between sister chromatids. DSB resection promotes recruitment of the Mec1 checkpoint kinase to the break. Rtt107 is a target of Mec1 and serves as a scaffold during repair. Rtt107 plays an important role during rescue of damaged replication forks, however whether Rtt107 contributes to the repair of DSBs is unknown. Here we show that Rtt107 is recruited to DSBs induced by the HO endonuclease. Rtt107 phosphorylation by Mec1 and its interaction with the Smc5-Smc6 complex are both required for Rtt107 loading to breaks, while Rtt107 regulators Slx4 and Rtt101 are not. We demonstrate that Rtt107 has an effect on the efficiency of sister chromatid recombination (SCR) and propose that its recruitment to DSBs, together with the Smc5-Smc6 complex is important for repair through the SCR pathway. PMID- 21647454 TI - Intercellular bridges in vertebrate gastrulation. AB - The developing zebrafish embryo has been the subject of many studies of regional patterning, stereotypical cell movements and changes in cell shape. To better study the morphological features of cells during gastrulation, we generated mosaic embryos expressing membrane attached Dendra2 to highlight cellular boundaries. We find that intercellular bridges join a significant fraction of epiblast cells in the zebrafish embryo, reaching several cell diameters in length and spanning across different regions of the developing embryos. These intercellular bridges are distinct from the cellular protrusions previously reported as extending from hypoblast cells (1-2 cellular diameters in length) or epiblast cells (which were shorter). Most of the intercellular bridges were formed at pre-gastrula stages by the daughters of a dividing cell maintaining a membrane tether as they move apart after mitosis. These intercellular bridges persist during gastrulation and can mediate the transfer of proteins between distant cells. These findings reveal a surprising feature of the cellular landscape in zebrafish embryos and open new possibilities for cell-cell communication during gastrulation, with implications for modeling, cellular mechanics, and morphogenetic signaling. PMID- 21647455 TI - 2011 Canadian Association of Gastroenterology educational needs assessment report. PMID- 21647456 TI - Fecal occult blood testing while waiting for screening colonoscopy in average risk individuals: durable option or short-term solution? PMID- 21647457 TI - Reactions to a targeted intervention to increase fecal occult blood testing among average-risk adults waiting for screening colonoscopy. AB - BACKGROUND: Increasing demand combined with limited capacity has resulted in long wait times for average-risk adults referred for screening colonoscopy for colorectal cancer. Management of patients on these growing wait lists is an emerging clinical issue. OBJECTIVE: To inform the content and design of a mailed targeted invitation for patients to undergo annual fecal occult blood testing (FOBT) while awaiting colonoscopy. METHODS: Focus groups (FGs) with average-risk patients on a wait list for screening colonoscopy at a high-throughput academic outpatient colonoscopy facility were conducted. During each FG session, feedback regarding a range of materials under consideration for the planned intervention was elicited using a semistructured facilitator guide. The FG sessions were recorded and transcribed verbatim, and analyzed using the constant comparative method to identify key themes. RESULTS: Findings from the three FGs (n=28) suggested that average risk patients on a wait list for screening colonoscopy would be receptive to a targeted intervention recommending they undergo FOBT while waiting. Participants indicated that the invitation to undergo FOBT was an important acknowledgement that they were on an actively managed list, and that a mechanism to ensure that they were correctly triaged while waiting was in place. Several specific suggestions to improve the design of the targeted intervention were obtained. CONCLUSIONS: Results of the present study provide useful information for developing effective strategies to manage average-risk individuals facing long wait times for screening colonoscopy. PMID- 21647458 TI - Sedation practices in Canada: a propos de propofol. PMID- 21647459 TI - A survey of sedation practices for colonoscopy in Canada. AB - BACKGROUND: There are limited data regarding the use of sedation for colonoscopy and concomitant monitoring practices in different countries. METHODS: A survey was mailed to 445 clinician members of the Canadian Association of Gastroenterology and 80 members of the Canadian Society of Colon and Rectal Surgeons in May and June 2009. RESULTS: Sixty-five per cent of Canadian Association of Gastroenterology members and 69% of Canadian Society of Colon and Rectal Surgeons members responded with the full survey. Most endoscopists reported using sedation for more than 90% of colonoscopies. The most common sedation regimen was a combination of midazolam and fentanyl. Propofol, either alone or with another drug, was used in 12% of cases. A higher proportion (94%) of adult gastroenterologists who routinely used propofol were highly satisfied compared with those using other sedative agents (45%; P<0.001). Fifty per cent of adult gastroenterologists and 29% of surgeons who were not currently using propofol expressed interest in starting to use it for routine colonoscopies. Only a single nurse was present in the endoscopy room during colonoscopy performed by two-thirds of the endoscopists. CONCLUSIONS: Results of the present survey suggest that gastroenterologists in Canada use sedation for colonoscopy in more than 90% of their patients. There was higher satisfaction among gastroenterologists who used propofol routinely for all colonoscopies. Most endoscopy rooms were staffed by a single nurse, which may limit further increases in the use of propofol. Further studies are needed to determine optimal staffing of endoscopy units with and without the use of propofol. Sedation practices of general surgery endoscopists need to be evaluated. PMID- 21647460 TI - Conventional versus Rosemont endoscopic ultrasound criteria for chronic pancreatitis: comparing interobserver reliability and intertest agreement. AB - BACKGROUND: The Rosemont criteria (RC) were recently proposed by expert consensus to standardize endoscopic ultrasound (EUS) features and thresholds for diagnosing chronic pancreatitis (CP); however, they are cumbersome and are not validated. OBJECTIVE: To determine interobserver agreement between RC and conventional criteria (CC), and to assess intertest agreement in the diagnosis of CP. METHODS: Thirty-six consecutive patients who underwent EUS for abdominal pain or pancreatitis were retrospectively reviewed. Anonymized images were independently chosen as best representations of the pancreatic body and reviewed by three experts who recorded the presence of CC and RC features. Agreement (proportion and kappa statistic) between CC and RC was calculated. Interobserver agreement within the CC and RC was assessed. Secondary comparisons with endoscopic retrograde cholangiopancreatography were made where available. RESULTS: Using CC, 60 readings (83.3%) were negative for CP, while 12 readings (16.7%) were positive. Using RC, 59 readings (81.9%) were negative for CP, while 13 (18.1%) were positive. The weighted kappa for interobserver agreement for CC (four categories: normal/low probability, indeterminate, high probability or calcific) was 0.50, with 80.0% overall agreement, versus 0.27 and 68.1% for the four RC categories (normal, indeterminate, suggestive of and consistent with). Agreement on a positive diagnosis with CC was 86.1% (P=0.38 [McNemar's exact test]), with a kappa of 0.47; for RC, agreement was lower at 80.6% (P=0.016 [McNemar's exact test]), with a kappa of 0.38. For patients who underwent endoscopic retrograde cholangiopancreatography (n=12), false-negative and false-positive rates between CC and RC did not appear to be different. CONCLUSIONS: The RC do not appear to achieve the goals of improving accuracy and interobserver agreement for diagnosing CP. PMID- 21647461 TI - Gastrostomy tube insertion in children: the Edmonton experience. AB - BACKGROUND: Although gastrostomy tube insertion - whether endoscopic or open - is generally safe, procedure-related complications have been reported. OBJECTIVE: To compare gastrostomy tube insertion-related complications between percutaneous endoscopic gastrostomy and open gastrostomy at a single pediatric centre. METHODS: The charts of children (younger than 17 years of age at the time of tube insertion) who underwent endoscopic or open gastrostomy tube insertion from January 2005 to December 2007 at the Stollery Children's Hospital (Edmonton, Alberta) were examined. RESULTS: A total of 298 children underwent gastrostomy tube insertion over a period of three years. After excluding patients with incomplete charts, 160 children (91 boys, mean [+/- SD] age 3.18 +/- 4.73 years) were included. Eighty-five children (mean age 4.50 +/- 5.40 years) had their gastrostomy tube inserted endoscopically, while the remaining 75 (mean age 1.68 +/- 3.27 years; P<0.001) underwent an open procedure. The overall rate of major complications was 10.2% for the endoscopic technique and 8.6% for the open technique (P=0.1). Major infections were higher in the endoscopic technique group, while persistent gastrocutaneous fistulas after tube removal were more common in the open technique group. CONCLUSION: Although the rate of major complications was similar between the endoscopic and open tube insertion groups, major infections were more common among children who underwent endoscopic gastrostomy. The decision for gastrostomy tube insertion was primarily based on clinical background. PMID- 21647462 TI - Assessing inflammatory bowel disease-associated antibodies in Caucasian and First Nations cohorts. AB - BACKGROUND: First Nation populations in Canada have a very low incidence of inflammatory bowel disease (IBD). Based on typical infections in this population, it is plausible that the First Nations react differently to microbial antigens with a different antibody response pattern, which may shed some light as to why they experience a low rate of IBD. OBJECTIVE: To compare the positivity rates of antibodies known to be associated with IBD in Canadian First Nations compared with a Canadian Caucasian population. METHODS: Subjects with Crohn's disease, ulcerative colitis (UC), rheumatoid arthritis (RA) (as an immune disease control) and healthy controls without a personal or family history of chronic immune diseases, were enrolled in a cohort study aimed to determine differences between First Nations and Caucasians with IBD or RA. Serum from a random sample of these subjects (n=50 for each of First Nations with RA, First Nations controls, Caucasians with RA, Caucasians with Crohn's disease, Caucasians with UC and Caucasians controls, and as many First Nations with either Crohn's disease or UC as could be enrolled) was analyzed in the laboratory for the following antibodies: perinuclear antineutrophil cytoplasmic antibody (pANCA), and four Crohn's disease-associated antibodies including anti-Saccharomyces cerevisiae, the outer membrane porin C of Escherichia coli, I2 - a fragment of bacterial DNA associated with Pseudomonas fluorescens, and the bacterial flagellin CBir-1. The rates of positive antibody responses and mean titres among positive results were compared. RESULTS: For pANCA, First Nations had a positivity rate of 55% in those with UC, 32% in healthy controls and 48% in those with RA. The pANCA positivity rate was 32% among Caucasians with RA. The rates of the Crohn's disease associated antibodies for the First Nations and Caucasians were comparable. Among First Nations, up to one in four healthy controls were positive for any one of the Crohn's disease-associated antibodies. First Nations had significantly higher pANCA titres in both the UC and RA groups than Caucasians. DISCUSSION: Although First Nation populations experience a low rate of IBD, they are relatively responsive to this particular antibody panel. CONCLUSIONS: The positivity rates of these antibodies in First Nations, despite the low incidence of IBD in this population, suggest that these antibodies are unlikely to be of pathogenetic significance. PMID- 21647463 TI - The metabolic syndrome is associated with complicated gallstone disease. AB - BACKGROUND: Gallstone disease (GD) is a common condition worldwide. Several studies demonstrated that the presence of gallstones is strongly associated with cardiovascular disease. The metabolic syndrome is a highly prevalent cardiovascular condition. OBJECTIVE: To examine the relationship between complicated GD (CGD) and the metabolic syndrome or its components. METHODS: Two hundred seventeen patients with gallstones were examined. All patients underwent biliary ultrasonography after a complete medical history and laboratory examination. Data collection for the diagnosis of metabolic syndrome included measurements of waist circumference, blood pressure and lipids, and biochemical tests. RESULTS: Of the 217 patients examined, 115 patients (53%) had CGD and 102 patients (47%) had uncomplicated GD (UCGD). There was a significant difference between the number of patients with large gallstones in the CGD and UCGD groups (n=14 [12%] versus n=2 [2%], respectively; P=0.004). Metabolic syndrome, diabetes mellitus and large waist circumference were more prevalent in the CGD group than in the UCGD group. Homeostatic model assessment of insulin resistance scores were higher in the CGD group than in UCGD group (2.51 [95% CI 0.57 to 23.90] versus 2.20 [95% CI 0.09 to 8.87], respectively; P=0.032). Logistic regression analysis revealed that the presence of metabolic syndrome (OR 1.434; 95% CI 1.222 to 1.846, P=0.014), diabetes mellitus (OR 1.493; 95% CI 1.255 to 1.953; P=0.035) and large gallstones (OR 1.153; 95% CI 1.033 to 1.714; P=0.017) were independent predictors of CGD. CONCLUSION: Results of the present study demonstrated that metabolic syndrome, diabetes and gallstone size were associated with CGD. Further prospective studies are needed to understand the clinical importance of this association. PMID- 21647464 TI - Interstitial cells of Cajal: a novel hypothesis for the pathophysiology of irritable bowel syndrome. AB - Irritable bowl syndrome (IBS) affects a large proportion of the world's population, and accounts for a considerable number of visits to gastroenterologists and general practitioners. Despite its high prevalence, the precise mechanism of IBS has not been identified to date. The interstitial cells of Cajal (ICC) participate in the production of slow waves and the regulation of their propagation through the gastrointestinal system; thus, they are important components of gastrointestinal motility. The present review proposes that ICC play a central role in the pathophysiology of IBS. This hypothesis is based on many links between ICC and currently proposed mechanisms of IBS pathophysiology. It appears that ICC may be involved in almost all of the previously explained pathogenic mechanisms of IBS. If proven, this hypothesis may provide a key to solving the IBS mystery. PMID- 21647465 TI - Acoustic neuroma: what the evidence says about evaluation and treatment. AB - Tumor size and a patient's age, condition, and personal preference are key factors in choosing among watchful waiting, microsurgery, or stereotactic radiosurgery. PMID- 21647466 TI - The perils of PSA screening. PMID- 21647467 TI - You wrote the prescription, but will it get filled? AB - PURPOSE: Despite numerous studies on adherence, there is little research on the first-fill rate of antihypertensive prescriptions. Our study took advantage of the recent increase in electronic prescribing (e-prescribing) and used data from e-prescribing physicians to determine the first-fill failure rate of antihypertensive prescriptions and to assess which factors predict first-fill failure. METHODS: This retrospective study reviewed claims from a Mid-Atlantic managed care organization (MCO). We included adult members with continuous medical and pharmacy coverage who were prescribed an antihypertensive in 2008 by an e-prescribing physician. First-fill failure occurred when the patient did not obtain the antihypertensive medication due to either a denial by the MCO or reversal by the dispensing pharmacist. (Pharmacists reverse claims when a patient fails to pick up a medication.) Multivariate regression analysis determined the clinical and demographic factors associated with failure to fill. RESULTS: The cohort consisted of 14,693 antihypertensive prescriptions, prescribed by 164 e prescribing physicians for 7061 unique members. There were 2289 out of 14,693 prescriptions (15.6%) that went unfilled, affecting 24.3% of patients. Of the prescriptions not obtained, 1466 (64%) were denied by the MCO and 823 (36%) were reversed. Significant factors associated with first-fill failure were new diagnosis of hypertension, new antihypertensive agent, higher co-payment, and enrollment in a health maintenance organization or preferred provider organization. CONCLUSIONS: Patients newly diagnosed with hypertension and those prescribed a new antihypertensive were at particularly high risk for not obtaining their medication. Because nearly a quarter of patients did not obtain their initial fill of an antihypertensive prescription, future research should determine efficient and cost-effective systems to address first-fill failure in primary care. PMID- 21647468 TI - "Doctor, my thumb hurts". AB - How should you treat these common causes of nontraumatic thumb pain? Use this evidence-based review as a clinical decision-making guide. PMID- 21647469 TI - Strategies for managing hot flashes. AB - Hormone therapy--at the lowest possible dose for the shortest period of time- remains the best option for menopausal women with moderate to severe vasomotor symptoms. PMID- 21647470 TI - When to suspect interstitial cystitis. AB - The symptom profile and comorbidities associated with this painful condition can make it difficult to diagnose--unless you know what to look for. PMID- 21647471 TI - PURLs: PSA testing: when it's useful, when it's not. AB - Routine PSA testing leads to more diagnoses of prostate cancer, but does not save lives. At least one group of men, however, may reap a small benefit. PMID- 21647472 TI - Unexpected skin necrosis of the thighs. AB - The patient was initially told she had cellulitis and sent home with a combination antibiotic. Three weeks later, she returned with necrotic lesions. PMID- 21647473 TI - Clinical Inquiry: What is the long-term educational outlook for youngsters with ADHD? AB - A prospective cohort study compared educational and employment outcomes among 91 middle-class white boys, 6 to 12 years of age, with ADHD who were referred to a psychiatric clinic with outcomes for 96 matched controls. Investigators used multiple educational achievement tests to evaluate participants when they enrolled in the study, then administered educational and occupational questionnaires 16 years later. Boys with ADHD completed 2.5 fewer years of school than controls (P=.001). Although rates of employment for the 2 groups were the same at 90%, those with ADHD had a significantly poorer occupational ranking than controls using the Hollingshead and Redlich system, which rates occupations on 7 point scale, with 1 representing top-ranked occupations. Individuals with ADHD scored 4.4 points compared with 3.5 points for the control group (P<.001). However, by the end of the study, more individuals with ADHD owned and operated their own businesses compared with controls (18% vs 5%; P<.01). PMID- 21647474 TI - Clinical Inquiry: How should we use the coronary artery calcium score to predict cardiovascular risk? AB - Most atherosclerotic lesions are calcified. The degree of calcification is proportional to the severity of atherosclerosis and can be quantified by the CAC score as measured by electron beam computed tomography (EBCT). PMID- 21647475 TI - An ab initio study of cooperative effects in ternary complexes X:CNH:Z with X, Z = CNH, FH, ClH, FCl, and HLi: structures, binding energies, and spin-spin coupling constants across intermolecular bonds. AB - A systematic ab initio investigation has been carried out to determine the structures, binding energies, and spin-spin coupling constants of ternary complexes X:CNH:Z and corresponding binary complexes X:CNH and CNH:Z, for X, Z = CNH, FH, ClH, FCl, and HLi. The enhanced binding energies of ternary complexes X:CNH:Z for fixed X as a function of Z decrease in the same order as the binding energies of the binary complexes CNH:Z. In contrast, the enhanced binding energies of the ternary complexes for fixed Z as a function of X do not decrease in the same order as the binding energies of the binary complexes X:CNH, a consequence of the increased stabilities of ternary complexes FCl:CNH:Z due to very strong chlorine-shared halogen bonds. For complexes in which the X...CNH interaction is a D-H...C hydrogen bond for D-H the proton-donor group (N-H, F-H, or Cl-H), spin-spin coupling constants (1)J(D-H) and (2h)J(D-C) in ternary complexes X:CNH:Z decrease in absolute value as the binding energies of binary complexes CNH:Z and the enhanced binding energies of the ternary complexes for fixed X as a function of Z also decrease. However, (2X)J(F-C) increases as the enhanced binding energies of the ternary complexes FCl:CNH:Z decrease, a consequence of the nature of the chlorine-shared halogen bond. The one-bond coupling constants (1)J(N-H) for the CNH...Z interaction in ternary complexes vary significantly, depending on the nature of the X...CNH interaction. The largest values of (1)J(N-H) are found for ternary complexes with FCl as X. Two bond coupling constants (2h)J(N-A) for A the proton-acceptor atom of Z, and (2d)J(N-H) decrease in absolute value in the order of decreasing enhancement energies of ternary complexes X:CNH:Z for fixed Z as a function of X. PMID- 21647476 TI - Pyrrolidinyl-sulfamide derivatives as a new class of bifunctional organocatalysts for direct asymmetric Michael addition of cyclohexanone to nitroalkenes. AB - A series of chiral pyrrolidinyl-sulfamide derivatives have been identified as efficient bifunctional organocatalysts for the direct Michael addition of cyclohexanone to a wide range of nitroalkenes. The desired Michael adducts were obtained in high chemical yields and excellent stereoselectivities (up to 99/1 dr and 95% ee). PMID- 21647477 TI - Radiolysis of astrophysical ice analogs by energetic ions: the effect of projectile mass and ice temperature. AB - An experimental study of the interaction of highly charged, energetic ions (52 MeV (58)Ni(13+) and 15.7 MeV (16)O(5+)) with mixed H(2)O : C(18)O(2) astrophysical ice analogs at two different temperatures is presented. This analysis aims to simulate the chemical and the physicochemical interactions induced by cosmic rays inside dense, cold astrophysical environments, such as molecular clouds or protostellar clouds as well at the surface of outer solar system bodies. The measurements were performed at the heavy ion accelerator GANIL (Grand Accelerateur National d'Ions Lourds) in Caen, France. The gas samples were deposited onto a CsI substrate at 13 K and 80 K. In situ analysis was performed by a Fourier transform infrared (FTIR) spectrometer at different fluences. Radiolysis yields of the produced species were quantified. The dissociation cross section at 13 K of both H(2)O and CO(2) is about 3-4 times smaller when O ions are employed. The ice temperature seems to affect differently each species when the same projectile was employed. The formation cross section at 13 K of molecules such as C(18)O, CO (with oxygen from water), and H(2)O(2) increases when Ni ions are employed. The formation of organic compounds seems to be enhanced by the oxygen projectiles and at lower temperatures. In addition, because the organic production at 13 K is at least 4 times higher than the value at 80 K, we also expect that interstellar ices are more organic-rich than the surfaces of outer solar system bodies. PMID- 21647478 TI - Large ferroelectric polarization in the new double perovskite NaLaMnWO6 induced by non-polar instabilities. AB - Based on density functional theory calculations and group theoretical analysis, we have studied NaLaMnWO(6) compound which has been recently synthesized [G. King, A. Wills and P. M. Woodward, Phys. Rev. B: Condens. Matter, 2009, 79, 224428] and belongs to the AA'BB'O(6) family of double perovskites. At low temperature, the structure has monoclinic P2(1) symmetry, with layered ordering of the Na and La ions and rocksalt ordering of Mn and W ions. The Mn atoms show an antiferromagnetic collinear spin ordering, and the compound has been reported as a potential multiferroic. By comparing the low symmetry structure with a parent phase of P4/nmm symmetry, two distortion modes are found dominant. They correspond to MnO(6) and WO(6) octahedron tilt modes, often found in many simple perovskites. While in the latter these common tilting instabilities yield non polar phases, in NaLaMnWO(6) the additional presence of the A-A' cation ordering is sufficient to make these rigid unit modes a source of the ferroelectricity. Through a trilinear coupling with the two unstable tilting modes, a polar distortion is induced, although the system has no intrinsic polar instability. The calculated electric polarization resulting from this polar distortion is as large as ~16 MUC cm(-2). Despite its secondary character, this polarization is coupled with the dominant tilting modes and its switching is bound to produce the switching of one of two tilts, enhancing in this way a possible interaction with the magnetic ordering. The transformation of common non-polar purely steric instabilities into sources of ferroelectricity through a controlled modification of the parent structure, as done here by the cation ordering, is a phenomenon to be further explored. PMID- 21647479 TI - Dimers of cyclic carbonates: chirality recognition in battery solvents and energy storage. AB - Dimers of ethylene carbonate and propylene carbonate are created in supersonic jet expansions and characterized by FTIR spectroscopy. Fermi resonances are switched on and off by dimerization. There is a unique centrosymmetric dimer of ethylene carbonate in a pronounced case of complementary chirality synchronization, contributing to its energy storage capacity at melting. Two chiral propylene carbonate molecules combine in more intricate ways. If they have the same handedness, one of them is forced into an axial conformation and the binding partner stays in the more stable equatorial structure. If they have opposite handedness, centrosymmetric dimers of either axial or equatorial conformations are formed. This suggests the usefulness of chirality control in elucidating ionic transport mechanisms in battery solvents and asymmetric catalysis in such solvents. PMID- 21647481 TI - para-Mercaptobenzoic acid-modified silver nanoparticles as sensing media for the detection of ammonia in air based on infrared surface enhancement effect. AB - To utilize the large signals provided by surface-enhanced infrared absorption (SEIRA) measurements for chemical sensing, a new sensing scheme was proposed and demonstrated for detection of ammonia in air samples. To increase the SEIRA effect, a sensing phase composed of multi-layers of silver nanoparticles (AgNPs) was prepared using a chemically controlled electroless deposition method. para Mercaptobenzoic acid (pMBA) served as the controlling agent in formation of AgNPs, a surface modification agent of AgNPs for sensing, and a stabilizer to protect the AgNPs from coagulation and oxidation. The sensing approach utilized the interaction between pMBA and ammonia, which involves the formation of carboxylate-ammonium complex. After interaction, the enhanced IR absorption bands of pMBA on AgNPs were significantly changed and able to provide quantitative information on the ammonia concentrations. To optimize the conditions for preparing sensing elements, parameters used to form multi-layers of AgNPs were systematically varied and their corresponding sensitivities in detection of ammonia were recorded. The results indicate that AgNPs with diameters in the range of 100 nm provided the best performance in terms of detecting ammonia via the SEIRA effect. Also, the analytical signal generally increased as the number of layers of AgNPs increased, but was limited to certain layers, depending on the reaction conditions used in preparation of AgNPs. The sensing elements were found to be highly selective to ammonia and the detection limit approached 150 ppb with a linear range up to 25 ppm. PMID- 21647480 TI - Metal centers in the anaerobic microbial metabolism of CO and CO2. AB - Carbon dioxide and carbon monoxide are important components of the carbon cycle. Major research efforts are underway to develop better technologies to utilize the abundant greenhouse gas, CO(2), for harnessing 'green' energy and producing biofuels. One strategy is to convert CO(2) into CO, which has been valued for many years as a synthetic feedstock for major industrial processes. Living organisms are masters of CO(2) and CO chemistry and, here, we review the elegant ways that metalloenzymes catalyze reactions involving these simple compounds. After describing the chemical and physical properties of CO and CO(2), we shift focus to the enzymes and the metal clusters in their active sites that catalyze transformations of these two molecules. We cover how the metal centers on CO dehydrogenase catalyze the interconversion of CO and CO(2) and how pyruvate oxidoreductase, which contains thiamin pyrophosphate and multiple Fe(4)S(4) clusters, catalyzes the addition and elimination of CO(2) during intermediary metabolism. We also describe how the nickel center at the active site of acetyl CoA synthase utilizes CO to generate the central metabolite, acetyl-CoA, as part of the Wood-Ljungdahl pathway, and how CO is channelled from the CO dehydrogenase to the acetyl-CoA synthase active site. We cover how the corrinoid iron-sulfur protein interacts with acetyl-CoA synthase. This protein uses vitamin B(12) and a Fe(4)S(4) cluster to catalyze a key methyltransferase reaction involving an organometallic methyl-Co(3+) intermediate. Studies of CO and CO(2) enzymology are of practical significance, and offer fundamental insights into important biochemical reactions involving metallocenters that act as nucleophiles to form organometallic intermediates and catalyze C-C and C-S bond formations. PMID- 21647486 TI - Quantification of changes in oxygen release from red blood cells as a function of age based on magnetic susceptibility measurements. AB - This study extends the in vitro understanding of the RBC storage lesion by serially analyzing the RBC's magneophoretic mobility, a property dependent on the content and oxygenation or oxidation state of hemoglobin (Hb) iron, during storage. Four prestorage leukoreduced, AS-5 preserved RBC units were stored between 1 and 6 degrees C for 42 days. Weekly starting on storage day 7, each unit was sampled, divided into 3 aliquots, each subjected to different reactions: one aliquot was exposed to room air to produce oxyhemoglobin (oxyHb), another aliquot was mixed with sodium nitrite to produce methemoglobin (metHb), while the third aliquot was desaturated of oxygen (deoxyhemoglobin, deoxyHb) using nitrogen gas. These aliquots were placed into a cell tracking velocimetry (CTV) apparatus which measured both the settling velocity (u(s)) of the RBCs as well as their magnetically induced velocity (u(m)). The u(m)/u(s) ratio depends on the oxygenation state of the hemoglobin and the quantity of iron within the RBC. The RBC density was measured by percoll centrifugation. There was a significant reduction in the u(m)/u(s) ratio for the deoxyHb RBC aliquot as storage time elapsed, with a smaller but still significant reduction in the u(m)/u(s) ratio for the metHb aliquot. The average RBC density decreased very slightly during storage, as determined by the percoll centrifugation technique, although the average settling velocity (another measure of cell density) seemed to fluctuate during storage. The decrease in magnetophoretic mobility of the deoxyHb portion is explicable either by Hb's increased affinity for oxygen during storage, or else a loss of iron from the cells. PMID- 21647487 TI - Entrapment of [Ru(bpy)3]2+ in the anionic metal-organic framework: novel photoluminescence behavior exhibiting dual emission at room temperature. AB - [Ru(bpy)(3)](2+) (bpy = 2,2'-bipyridine) ions were entrapped into the cavities of two-dimensional anionic sheet-like coordination polymeric networks of [M(dca)(3)](-) (dca = dicyanamide; M = Mn(II) and Fe(II)). The prepared compounds, {[Ru(bpy)(3)][Mn(dca)(3)](2)}(n) (1) and {[Ru(bpy)(3)][Fe(dca)(3)](2)}(n) (2), were structurally characterized by X-ray single crystal analysis. The spectroscopic properties of the [Ru(bpy)(3)](2+) ion dramatically changed on its entrapment in [M(dca)(3)](-). The [Ru(bpy)(3)](2+) moiety present in 1 and 2 exhibits novel dual photo-emission at room temperature. PMID- 21647488 TI - Fabricating protein immunoassay arrays on nitrocellulose using dip-pen lithography techniques. AB - Advancements in lithography methods for printing biomolecules on surfaces are proving to be potentially beneficial for disease screening and biological research. Dip-pen nanolithography (DPN) is a versatile micro and nanofabrication technique that has the ability to produce functional biomolecule arrays. The greatest advantage, with respect to the printing mechanism, is that DPN adheres to the sensitive mild conditions required for biomolecules such as proteins. We have developed an optimised, high-throughput printing technique for fabricating protein arrays using DPN. This study highlights the fabrication of a prostate specific antigen (PSA) immunoassay detectable by fluorescence. Spot sizes are typically no larger than 8 MUm in diameter and limits of detection for PSA are comparable with a commercially available ELISA kit. Furthermore, atomic force microscopy (AFM) analysis of the array surface gives great insight into how the nitrocellulose substrate functions to retain protein integrity. This is the first report of protein arrays being printed on nitrocellulose using the DPN technique and the smallest feature size yet to be achieved on this type of surface. This method offers a significant advance in the ability to produce dense protein arrays on nitrocellulose which are suitable for disease screening using standard fluorescence detection. PMID- 21647489 TI - Photocatalytic hydrogen production. AB - The efficient storage of solar energy in chemical fuels, such as hydrogen, is essential for the large-scale utilisation of solar energy systems. Recent advances in the photocatalytic production of H(2) are highlighted. Two general approaches for the photocatalytic hydrogen generation by homogeneous catalysts are considered: HX (X = Cl, Br) splitting involving both proton reduction and halide oxidation via an inner-sphere mechanism with a single-component catalyst; and sensitized H(2) production, employing sacrificial electron donors to regenerate the active catalyst. Future directions and challenges in photocatalytic H(2) generation are enumerated. PMID- 21647490 TI - Physical analysis of the through-ligand long-distance magnetic coupling: spin polarization versus Anderson mechanism. AB - The physical factors governing the magnetic coupling between two magnetic sites are analyzed and quantified as functions of the length of the bridging conjugated ligand. Using wave-function-theory based ab initio calculations, it has been possible to separate and calculate the various contributions to the magnetic coupling, i.e. the direct exchange, the spin polarization and the kinetic exchange. It is shown in model systems that while the Anderson mechanism brings the leading contribution for short-length ligands, the spin polarization dominates the through-long-ligand couplings. Since the spin polarization decreases more slowly than the kinetic exchange, highly spin polarizable bridging ligands would generate a good magneto-communication between interacting magnetic units. PMID- 21647491 TI - Hydrogen bond formation of formamide and N-methylformamide in aqueous solution studied by quantum mechanical charge field-molecular dynamics (QMCF-MD). AB - The formation of hydrogen-bonds of formamide and "cis"-N-methylformamide in aqueous solution was examined using double zeta HF level Quantum Mechanical Charge Field-Molecular Dynamics (QMCF-MD) simulations. Basic attributes such as structure and dynamics of the solvates and hydrogen-bonds were studied in particular by means of coordination number distributions, mean residence times and radial distribution functions, on which spatial restrictions in the form of planes and cones were applied. Advanced methods of analysis gave detailed information about the sterical environment and the dynamic behavior of strong and weak hydrogen-bonds formed by the residues. The comparison of both molecules over a sampling period of 12 ps provided information on the influence of methylation of the amide function on molecular and hydration properties. PMID- 21647492 TI - Dissociative electron attachment to gas-phase formamide. AB - Dissociative electron attachment (DEA) to gaseous formamide, HCONH(2), has been investigated in the energy range between 0 eV and 18 eV using a crossed electron/molecule beam technique. The negative ion fragments have been comprehensively monitored and assigned to molecular structures by comparison with the results for two differently deuterated derivatives, namely 1D-formamide, DCONH(2), and N,N,D-formamide, HCOND(2). The following products were observed: HCONH(-), CONH(2)(-), HCON(-), OCN(-), HCNH(-), CN(-), NH(2)(-)/O(-), NH(-), and H(-). NH(2)(-) was also separated from O(-) by using high-resolution negative ion mass spectrometry. Four resonant dissociation channels can be resolved, the strongest ones being located between 2.0 and 2.7 eV and between 6.0 and 7.0 eV. CN(-) as the most abundant fragment and HCONH(-) are the dominant products of the first of these two resonances. The most important products of the latter resonance are NH(2)(-), CN(-), H(-), CONH(2)(-), and OCN(-). It is thus found that the loss of neutral H is a site-selective process, dissociation from the N site taking place between 2.0 and 2.7 eV while dissociation from the C site occurs between 6.0 and 7.0 eV. The suitability of these reactions and thus of formamide as an agent for electron-induced surface functionalisation is discussed. PMID- 21647493 TI - The oligodeoxynucleotide probes for the site-specific modification of RNA. AB - As the knowledge of the biological functions of RNA expands, the demand for research tools to investigate intracellular RNA is increasing. Oligonucleotides can be rationally designed for the target RNA sequence, and therefore, have become a reliable platform for the development of specific molecules for RNA. The chemical modification of RNA has a strong impact on RNA research; the fluorescent labeling of RNA is useful to monitor RNA production, processing, relocation in the cell, interaction with other intracellular components and degradation, etc. Chemical modification may affect the RNA function through a variety of pathways, and therefore, would be potentially useful for biological research, therapeutic approach and artificial manipulation of the RNA function. This tutorial review starts with an introduction of the biological relevance of modified RNA, and focuses on the recent progress of the oligodeoxynucleotide probes for the covalent modifications of RNA. The prospects of this new technology are also discussed. PMID- 21647494 TI - Formation of PbS materials from lead xanthate precursors. AB - Six lead xanthate adducts Pb(S(2)COR)(2).L [R = Et, (n)Bu, L = bipy, TMEDA (tetramethylethylenediamine), PMDETA (pentamethyldiethylenetriamine)] have been synthesised and the structures of all, save Pb(S(2)COBu(n))(2).TMEDA (4) which is an oil, determined. Pb(S(2)COEt)(2).TMEDA (3) is seven-coordinate at lead through three chelating ligands and one weak intermolecular Pb..S interaction. Both Pb(S(2)COR)(2).bipy [R = Et (1), (n)Bu (2)] are dimers in which one xanthate is terminal and the other MU(2) bridging at each sulphur, generating an eight coordinate lead when the bipy donor is included. Both Pb(S(2)COR)(2).PMDETA [R = Et (5), (n)Bu (6)] are seven-coordinate at lead by virtue of two bidentate chelating xanthate ligands and a tridentate PMDETA; there are no intermolecular interactions. Trends in the (207)Pb NMR chemical shifts mirror the changes in the intramolecular coordination number across the series. Pb(S(2)COEt)(2).TMEDA (3) has been used to deposit PbS films on glass, Mo-coated glass and Si by AACVD. Pb(S(2)COEt)(2) also generated PbS nanocubes when decomposed under an autogenerated pressure. PMID- 21647495 TI - Tri- and tetracoordinate copper(I) complexes bearing bidentate soft/hard SN and SeN ligands based on 2-aminopyridine. AB - The coordination properties of the EN ligands N-(2-pyridinyl)amino diphenylphosphine sulfide, N-(2-pyridinyl)amino-diisopropylphosphine sulfide, N (2-pyridinyl)amino-diphenylphosphine selenide, N-(2-pyridinyl)amino diisopropylphosphine selenide towards copper(I) precursors CuX (X = Br, I), [Cu(IPr)Cl] (IPr = 1,3-bis(2,6-diisopropylphenyl)imidazol-2-ylidene), and [Cu(CH(3)CN)(4)]PF(6) were studied. Treatment of CuX with EN ligands resulted in the formation of tricoordinate complexes of the type [Cu(kappa(2)(E,N)-EN)X]. The reaction of [Cu(IPr)Cl] with EN ligands, followed by halide abstraction with AgSbF(6), afforded cationic tricoordinate complexes [Cu(kappa(2)(S,N) EN)(IPr)](+), while the reaction of [Cu(CH(3)CN)(4)](+) with two equivalents of EN ligands yielded tetrahedral complexes [Cu(kappa(2)(E,N)-EN)(2)](+). Halide removal from [Cu(kappa(2)(S,N)-SN)I] with silver salts in the presence of L = CH(3)CN and CNtBu afforded dinuclear complexes of the type [Cu(kappa(2)(S,N),MU(S)-SN)(L)](2)(2+) containing bridging SN ligands. With the terminal alkynes HC=CC(6)H(4)Me and HC=CC(6)H(4)OMe, complexes of the formula [Cu(kappa(2)(S,N)-SN-iPr)(eta(2)-HC=CC(6)H(4)Me)](+) and [Cu(kappa(2)(S,N)-SN iPr)(eta(2)-HC=CC(6)H(4)OMe)](+) were obtained. The mononuclear nature of these compounds was supported by DFT calculations. Most complexes were also characterized by X-ray crystallography. PMID- 21647496 TI - Size-controlled synthesis of monodispersed mesoporous alpha-alumina spheres by a template-free forced hydrolysis method. AB - We demonstrated the formation of monodispersed spherical aluminum hydrous oxide precursors with tunable sizes by controlling the variables of a forced hydrolysis method. The particle sizes of aluminum hydrous oxide precursors were strongly dependent on the molar ratio of the Al(3+) reactants (sulfates and nitrates). In addition, the systematic phase and morphological evolutions from aluminum hydrous oxide to gamma-alumina (Al(2)O(3)) and finally to alpha-Al(2)O(3) through thermal dehydrogenation were characterized by X-ray powder diffraction (XRD), field emission scanning electron microscopy (FESEM), and transmission electron microscopy (TEM). After annealing the amorphous aluminum hydrous oxide in air at 900 degrees C and 1100 degrees C for 1 h, we observed complete conversion to phase-pure gamma- and alpha-Al(2)O(3), respectively, while maintaining monodispersity (125 nm, 195 nm, 320 nm, and 430 nm diameters were observed). Furthermore, both gamma- and alpha-Al(2)O(3) were found to be mesoporous in structure, providing enhanced specific surface areas of 102 and 76 m(2) g(-1), respectively, based on the Brunauer-Emmett-Teller (BET) measurement. PMID- 21647497 TI - A microfluidic platform for high-sensitivity, real-time drug screening on C. elegans and parasitic nematodes. AB - This paper describes a new microfluidic platform for screening drugs and their dose response on the locomotion behavior of free living nematodes and parasitic nematodes. The system offers a higher sensitivity drug screening chip which employs a combination of existing and newly developed methods. Real-time observation of the entire drug application process (i.e. the innate pre-exposure locomotion, the transient response during drug exposure and the time-resolved, post-exposure behavior) at a single worm resolution is made possible. The chip enables the monitoring of four nematode parameters (number of worms responsive, number of worms leaving the drug well, average worm velocity and time until unresponsiveness). Each parameter generates an inherently different dose response; allowing for a higher resolution when screening for resistance. We expect this worm chip could be used as a robust cross species, cross drug platform. Existing nematode motility and migration assays do not offer this level of sophistication. The device comprises two principal components: behavioral microchannels to study nematode motility and a drug well for administering the dose and observing drug effects as a function of exposure time. The drug screening experiment can be described by three main steps: (i) 'pre-exposure study'- worms are inserted into the behavioral channels and their locomotion is characterized, (ii) 'dose exposure'- worms are guided from the behavioral microchannels into the drug well and held for a predefined time, during which time their transient response to the dose is characterized and (iii) 'post exposure study'- worms are guided back into the behavioral microchannels where their locomotion (i.e. their time-resolved response to the dose) is characterized and compared to pre-exposure motility. The direction of nematodes' movement is reliably controlled by the application of an electric field within a defined range. Control experiments (e.g. in the absence of any drug) confirm that the applied electric fields do not affect the worms' motility or viability. We demonstrate the workability of the microfluidic platform on free living Caenorhabditis elegans (wild-type N2 and levamisole resistant ZZ15 lev-8) and parasitic Oesophagotomum dentatum (levamisole-sensitive, SENS and levamisole resistant, LEVR) using levamisole (a well-studied anthelmintic) as the test drug. The proposed scheme of drug screening on a microfluidic device is expected to significantly improve the resolution, sensitivity and data throughput of in vivo testing, while offering new details on the transient and time-resolved exposure effects of new and existing anthelmintics. PMID- 21647498 TI - Distinguishing drug-induced minor morphological changes from major cellular damage via label-free impedimetric toxicity screening. AB - We present a novel perfusion-based microfluidic platform for label-free drug toxicity screening which can single out non-lethal morphological changes from cellular death using electrical impedance spectroscopy. Minor cellular changes such as cell-cell contacts and major cell injury were identified via impedance phase angle analysis and follow-up of impedance magnitude at different frequencies. Having exposed HepG2/C3A cells to acetaminophen (AP), we showed that continuous drug perfusion caused a time and concentration-dependent impedance decrease. Moreover, perfusion of repeated doses revealed altered dielectric properties of the cell culture after recovery from AP exposure. This study highlights the possibility to sense cellular changes long before cellular death takes place, pointing out the remarkable sensitivity advantage of this technique over standard endpoint viability tests and its interest for toxicology. PMID- 21647499 TI - Low temperature solution-phase growth of ZnSe and ZnSe/CdSe core/shell nanowires. AB - High quality ZnSe nanowires (NWs) and complementary ZnSe/CdSe core/shell species have been synthesized using a recently developed solution-liquid-solid (SLS) growth technique. In particular, bismuth salts as opposed to pre-synthesized Bi or Au/Bi nanoparticles have been used to grow NWs at low temperatures in solution. Resulting wires are characterized using transmission electron microscopy and possess mean ensemble diameters between 15 and 28 nm with accompanying lengths ranging from 4-10 MUm. Subsequent solution-based overcoating chemistry results in ZnSe wires covered with CdSe nanocrystals. By varying the shell's growth time, different thicknesses can be obtained and range from 8 to 21 nm. More interestingly, the mean constituent CdSe nanocrystal diameter can be varied and results in size-dependent shell emission spectra. PMID- 21647500 TI - Stable dye-labelled oligonucleotide-nanoparticle conjugates for nucleic acid detection. AB - Metallic nanoparticles functionalized with oligonucleotides are used for a number of nucleic acid detection strategies. However, oligonucleotide-nanoparticle conjugates suffer from a lack of stability when exposed to certain conditions associated with DNA detection assays. In this study, we report the synthesis of thiol and thioctic acid-modified oligonucleotide gold nanoparticle (OGNs) conjugates functionalized with a dye label and varying spacer groups. The thioctic acid-modified conjugates exhibit increased stability when treated with dithiothreitol (DTT) compared to the more commonly used thiol modification. When the dye labelled oligonucleotide nanoparticle conjugates are exposed to the same conditions there is a pronounced increase in the stability for both thioctic acid and thiol modified sequences. These results open up the possibility of simply using a dye label to enhance the stability of oligonucleotide-nanoparticle conjugates in DNA detection assays where the enhanced stability of the conjugate system can be advantageous in more complex biological environments. PMID- 21647501 TI - Palladium-catalyzed three-component domino reaction for the preparation of benzo[b]thiophene and related compounds. AB - A simple and efficient palladium-catalyzed three-component domino reaction of bromothiophenes with internal alkynes has been developed to produce benzo[b]thiophenes in moderate to good yields. PMID- 21647502 TI - Exploring the dark side of MTT viability assay of cells cultured onto electrospun PLGA-based composite nanofibrous scaffolding materials. AB - One major method used to evaluate the biocompatibility of porous tissue engineering scaffolding materials is MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide) assay. The MTT cell viability assay is based on the absorbance of the dissolved MTT formazan crystals formed in living cells, which is proportional to the number of viable cells. Due to the strong dye sorption capability of porous scaffolding materials, we propose that the cell viability determined from the MTT assay is likely to give a false negative result. In this study, we aim to explore the effect of the adsorption of MTT formazan on the accuracy of the viability assay of cells cultured onto porous electrospun poly(lactic-co-glycolic acid) (PLGA) nanofibers, HNTs (halloysite nanotubes)/PLGA, and CNTs (multiwalled carbon nanotubes)/PLGA composite nanofibrous mats. The morphology of electrospun nanofibers and L929 mouse fibroblasts cultured onto the nanofibrous scaffolds were observed using scanning electron microscopy. The viability of cells proliferated for 3 days was evaluated through the MTT assay. In the meantime, the adsorption of MTT formazan onto the same electrospun nanofibers was evaluated and the standard concentration absorbance curve was obtained in order to quantify the contribution of the adsorbed MTT formazan during the MTT cell viability assay. We show that the PLGA, and the HNTs- or CNTs-doped PLGA nanofibers display appreciable MTT formazan dye sorption, corresponding to 35.6-50.2% deviation from the real cell viability assay data. The better dye sorption capability of the nanofibers leads to further deviation from the real cell viability. Our study gives a general insight into accurate MTT cytotoxicity assessment of various porous tissue engineering scaffolding materials, and may be applicable to other colorimetric assays for analyzing the biological properties of porous scaffolding materials. PMID- 21647503 TI - Electrochemical immunoassay platform for high sensitivity protein detection based on redox-modified carbon nanotube labels. AB - We report a highly sensitive immunoassay protocol based on the use of redox modified multi-walled carbon nanotubes (MWNTs) as electrochemical labels. The MWNTs were coated with methylene blue (MB) at an optically-determined loading of 3.41 * 10(-3) mol g(-1), and were then attached to secondary antibodies (Ab(2)) by adsorption. As a model analyte mouse IgG was collected by primary antibody (Ab(1))-coated magnetic beads. Following binding of the MB-MWNT-Ab(2) conjugates, IgG could be measured by MB reduction. Using differential pulse voltammetry for quantification, IgG was calibrated with a dynamic range of 0.1 pg mL(-1) to 100 pg mL(-1). Given the different possible Ab(1)-MB-MWNT-Ab(2) orientations on the magnetic beads, it was likely that not all the MB communicated with the electrode. A greater quantity of MB could be accessed by using the Fe(CN)(6)(3-/4 ) redox couple as a solution phase mediator. This enabled us to lower the dynamic range down to 5 fg mL(-1) to 100 fg mL(-1). PMID- 21647504 TI - Evaluation of coated QCM for the detection of atmospheric ozone. AB - A coated acoustic wave sensor has been developed to selectively detect atmospheric ozone. The selective detection has been assessed using a variety of coatings: beeswax, gallic acid, indigo carmine, polybutadiene, potassium iodide and sodium nitrite. Polybutadiene was the most sensitive with a limit of detection of 55 ppb. The sensitivity was improved by operating at higher harmonics and was shown to increase linearly with harmonic up to the 11th harmonic. This novel work shows that ozone detection can be improved by operating at the crystal's harmonic frequencies and in conjunction with a suitable flow rate, a potentially highly sensitive and fast response sensor can be created based on acoustic wave technology. PMID- 21647505 TI - Subcellular localization of early biochemical transformations in cancer-activated fibroblasts using infrared spectroscopic imaging. AB - The tumor microenvironment, or stroma, is chemically and morphologically modified during carcinoma progression. The predominant cell type in the stroma, the fibroblast, maintains collagen properties in normal tissue and often transformed during tumor progression. Biochemical changes within fibroblasts upon initial cancer activation, however, are relatively poorly defined. Here, we hypothesized that Fourier transform infrared (FT-IR) spectroscopic imaging could potentially be employed to examine these early transformations. Further, we employ attenuated total reflectance (ATR) microscopy to characterize subcellular spectra and their changes upon transformation. We characterized fibroblast transitions upon stimulation with both a molecular agent and a carcinoma-mimicking cellular co culture system. Changes were predominantly observed in the 1080 cm(-1) and 1224 cm(-1) peak absorbance, commonly associated with nucleic acids, as well as in the band at 2930 cm(-1) associated with the C-H stretching of proteins in the cytoplasmic compartment. In conclusion, biochemical changes in cancer-associated fibroblasts that express alpha-SMA are dominated by the cytoplasm, rather than the nucleus. This ensures that spectral changes are not associated with proliferation or cell cycle processes of the cells and the cells are undergoing a true phenotypic change denoted by protein modifications in the cell body. PMID- 21647506 TI - A magnetically active microfluidic device for chemiluminescence bioassays. AB - Highly active horseradish peroxidase functionalized magnetic nanoparticles were prepared and packed into a microfluidic channel, producing an in-line bioreactor that enabled a sensitive chemiluminescence assay of H(2)O(2). The proposed magnetically active microfluidic device proved useful for chemiluminescence assays of biomedically interesting compounds. PMID- 21647507 TI - Synthesis, characterization, reactivity and catalytic activity of oxidovanadium(IV), oxidovanadium(V) and dioxidovanadium(V) complexes of benzimidazole modified ligands. AB - The reaction between [V(IV)O(acac)(2)] and the ONN donor Schiff base obtained by the condensation of pyridoxal and 2-aminoethylbenzimidazole (Hpydx-aebmz, I) or 2 aminomethylbenzimidazole (Hpydx-ambmz, II) in equimolar amounts results in the formation of [V(IV)O(acac)(pydx-aebmz)] 1 and [V(IV)O(acac)(pydx-ambmz)] 2, respectively. The aerobic oxidation of the methanolic solution of 1 yielded [V(V)O(2)(pydx-aebmz)] 3 and its reaction with aqueous H(2)O(2) gave the oxidoperoxidovanadium(v) complex, [V(V)O(O(2))(pydx-aebmz)] 4. The formation of 4 in solution is also established by titrations of methanolic solutions of 1 with H(2)O(2). By titrating solutions of 3 and of 4 with aqueous H(2)O(2) several distinct V(V)-pydx-aebmz species also containing the peroxido ligand are detected. The full geometry optimization of all species envisaged was done using DFT methods for suitable model complexes. The (51)V NMR chemical shifts (delta(V)) have also been calculated, the theoretical data being used to support assignments of the experimental chemical shifts. The (51)V hyperfine coupling constants are calculated for 1, the obtained values being in good agreement with the experimental EPR data. Reaction between the V(IV)O(2+) exchanged zeolite-Y and Hpydx-aebmz and Hpydx-ambmz in refluxing methanol, followed by aerial oxidation results in the formation of the encapsulated V(V)O(2)-complexes, abbreviated herein as [V(V)O(2)(pydx-aebmz)]-Y 5 and [V(V)O(2)(pydx-ambmz)]-Y 6. The molecular structure of 1, determined by single crystal X-ray diffraction, confirms its distorted octahedral geometry with the ONN binding mode of the tridentate ligand, with one acetylacetonato group remaining bound to the V(IV)O centre. Oxidation of styrene is investigated using some of these complexes as catalyst precursors with H(2)O(2) as oxidant. Under optimised reaction conditions for the conversion of styrene in acetonitrile, a maximum of 68% conversion of styrene (with [V(V)O(2)(pydx-aebmz)]-Y) and 65% (with [V(V)O(2)(pydx-ambmz)]-Y) is achieved in 6 h of reaction time. The selectivity of the various products is similar for both catalysts and follows the order: benzaldehyde (ca. 55%) > 1 phenylethane-1,2-diol > benzoic acid > styrene oxide > phenyl acetaldehyde. Speciation of the systems and plausible intermediates involved in the catalytic oxidation processes are established by UV-Vis, EPR, (51)V NMR and DFT studies. Both non-radical (Sharpless) and radical mechanisms of the olefin oxidations were theoretically studied, and the radical pathway was found to be even more favorable than the Sharpless mechanism. PMID- 21647508 TI - Equilibrium study on the interaction of phytic acid with polyamines and metal ions. AB - Interaction of phytic acid (myo-inositolhexakisphosphoric acid, IP) and polyamines (A = en, tn, Put, dien, 2,3-tri, 3,3-tri, Spd, 3,3,3-tet, spermine(Spm)) have been studied by potentiometric and (31)P-NMR techniques. The non-covalent interactions have led to the formation of stable molecular complexes of (IP)H(n)(A) type at the 1:1 molar ratio of the ligands, but of different numbers of protons. The IP protonation constants, stability constants of the molecular complexes and metal (Mg(2+)) complexes have been determined. The structural and pH dependences of stability constants showed the interactions between IP and A have the acid-base character determining their effectiveness, although the IP structure (5ax1eq, 5eq1ax) in molecular complexes should be also taken into account. (31)P NMR study showed in the presence of Spm (31)P highfield shifts and high pH shift of signal broadening due to chemical exchange between 5ax1eq and 5eq1ax. The preferable binding of Spm to IP over Mg(2+) in neutral pH indicated the importance of polyamine as a stabilizer of phosphate compounds. PMID- 21647509 TI - Stamping oriented molecular monolayers using liquid crystal inks. AB - Thermotropic liquid crystal (LC)-based inks are combined with patterned anchoring stamps to deposit organic monolayer films with simultaneous control over positional and molecular orientational order in a single step. PMID- 21647510 TI - Quantification of surface functional groups on polymer microspheres by supramolecular host-guest interactions. AB - We introduce a method to determine the number of accessible functional groups on a polymer microsphere surface based on the interaction between the macrocyclic host cucurbit[7]uril (CB7) and a guest reacted to the microsphere surface. After centrifugation, CB7 in the supernatant is quantified by addition of a fluorescent dye. The difference between added and detected CB7 affords the number of accessible surface functional groups. PMID- 21647511 TI - A platinum Chugaev carbene complex as a potent anticancer agent. AB - A platinum Chugaev complex was synthesised and fully characterized by multinuclear NMR spectroscopy and X-ray crystallography. This cis bis acyclic diamino carbene complex acts as a cytotoxic compound and behaves as a cisplatin equivalent by interacting with supercoiled DNA and thiols. Stability of the ligand is also discussed. PMID- 21647512 TI - Efficient electrocatalytic hydrogen production from H+ ions using specially designed boron-capped cobalt clathrochelates. AB - Specially designed hexachlorine-containing cobalt(II) tris-dioximate clathrochelates were found to efficiently electrocatalyze the production of molecular hydrogen from H(+) ions without the overpotential of this process. PMID- 21647513 TI - Lanthanide discs chill well and relax slowly. AB - The synthesis, structure and magnetic properties of two isostructural heptametallic lanthanide discs are reported, showing single molecule magnet (SMM) behaviour with a large energy barrier for the dysprosium analogue and a large magnetocaloric effect (MCE) for the gadolinium analogue. PMID- 21647514 TI - C. elegans metallothioneins: response to and defence against ROS toxicity. AB - The genome of the nematode Caenorhabditis elegans encodes for two multifunctional metal binding metallothioneins (MTs), CeMT-1 and CeMT-2. Here we applied qPCR to identify a transcriptional up-regulation following the exposure to free radical generators (ROS) paraquat or hydrogen peroxide, a trend that was confirmed with Pmtl::GFP expressing alleles. The deletion of the MT loci resulted in an elevation of in vivo levels of hydrogen peroxide and exposure to ROS caused a reduction in total egg production, growth and life span in wild type nematodes, effects that were particularly pronounced in the CeMT-2 and double knockout. Moreover, in vitro incubation of recombinant MTs with hydrogen peroxide demonstrated the presence of direct oxidation of the CeMTs, with zinc released from both isoforms and concomitant formation of intra-molecular disulfides. Finally, metabolic profiling (metabolomic) analysis of wild type and MT knockouts in the presence/absence of oxidative stressors, confirmed the overall trend described by the other experiments, and identified 2-aminoadipate as a potential novel small-molecule marker of oxidative stress. In summary, this study highlights that C. elegans metallothioneins scavenge and protect against reactive oxygen species and potentially against oxidative stress, with CeMT-2 being more effective than CeMT-1. PMID- 21647515 TI - A Bowman-Birk inhibitor with anti-elastase activity from Lathyrus sativus L. seeds. AB - Four Bowman-Birk inhibitors, named LSI-1/4, were isolated and purified from Lathyrus sativus L. seeds. The purification procedure consisted of two cation exchange chromatography steps, followed by gel-filtration and RP-HPLC. Mass spectrometry analysis of LSI-1/4 inhibitors yielded relative molecular masses of 7914.41 for LSI-1, 6867.67 for LSI-2, 7341.24 for LSI-3 and 7460.01 for LSI-4. N terminal sequences (up to 30 residues) of LSI-1/4 inhibitors were identical with the exception of sequence positions 21, 27 and 28 and highly similar to those of other Bowman-Birk inhibitors isolated from Leguminosae plants. Inhibitors LSI-1/4 were active towards trypsin and alpha-chymotrypsin, with IC(50) values for 12.6 nM of trypsin ranging from 4.9 to 24.3 nM. A lower activity was observed against bovine alpha-chymotrypsin (IC(50) values ranging from 0.5 to 3.4 MUM for 15.0 nM of alpha-chymotrypsin). Peptide mapping of the LSI-1 sequence showed the presence of an Ala residue in the second reactive site, thus explaining the low anti chymotrypsin activity of this inhibitor. In addition, LSI-1 was endowed with anti elastase activity, being able to inhibit human leukocyte elastase. PMID- 21647516 TI - Theoretical studies on the inactivation mechanism of gamma-aminobutyric acid aminotransferase. AB - The inactivation mechanism of gamma-aminobutyric acid aminotransferase (GABA-AT) in the presence of gamma-vinyl-aminobutyric acid, an anti-epilepsy drug, has been studied by means of theoretical calculations. Density functional theory methods have been applied to compare the three experimentally proposed inactivation mechanisms (Silverman et al., J. Biol. Chem., 2004, 279, 363). All the calculations were performed at the B3LYP/6-31+G(d,p) level of theory. Single point solvent calculations were carried out in water, by means of an integral equation formalism-polarizable continuum model (IEFPCM) at the B3LYP/6-31+G(d,p) level of theory. The present calculations provide an insight into the mechanistic preferences of the inactivation reaction of GABA-AT. The results also allow us to elucidate the key factors behind the mechanistic preferences. The computations also confirm the importance of explicit water molecules around the reacting center in the proton transfer steps. PMID- 21647517 TI - Ligand-free highly effective iron/copper co-catalyzed formation of dimeric aryl ethers or sulfides. AB - Highly selective coupling of diiodoarenes with phenols or phenthiols can be performed by using a low-cost, benign character and readily available Fe/Cu catalytic system in the absence of ligands. It is noteworthy that the desired dimeric aryl ethers or sulfides could be obtained in high yields by coupling between diiodoarenes and phenols, or diphenols with aryl iodides. PMID- 21647518 TI - Chlorination and ortho-acetoxylation of 2-arylbenzoxazoles. AB - Efficient and facile catalytic protocols for chlorination and ligand-directed ortho-acetoxylation of 2-arylbenzoxazoles have been developed. The chlorination is not a ligand-directed ortho-functionalization, but an electrophilic substitution process in the benzo ring of the benzoxazole moiety. Meanwhile, the acetoxylation exhibited high regioselectivity for the substrates containing a meta-substituent and occurred at the less sterically hindered ortho-C-H bond of the directing group. PMID- 21647519 TI - Copper-catalyzed direct thiolation of azoles with aliphatic thiols. AB - Cu(II)-catalyzed direct thiolation of azoles with thiols is described via intermolecular C-S bond formation/C-H functionalization under oxidative conditions. Both aryl thiols and aliphatic thiols are used as coupling partners, and furnished the thiolation products in moderate to good yields. The reaction is compatible with a wide range of heterocycles including oxazole, thiazole, imidazole and oxadiazole. PMID- 21647520 TI - Absolute stereochemistry and preferred conformations of urate degradation intermediates from computed and experimental circular dichroism spectra. AB - The enzymatic oxidation of urate leads to the sequential formation of optically active intermediates with unknown stereochemistry: (-)-5-hydroxyisourate (HIU) and (-)-2-oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline (OHCU). In accordance with the observation that a defect in HIU hydrolase causes hepatocarcinoma in mouse, a detoxification role has been proposed for the enzymes accelerating the conversion of HIU and OHCU into optically active (+)-allantoin. The enzymatic products of urate oxidation are normally not present in humans, but are formed in patients treated with urate oxidase. We used time-dependent density functional theory (TDDFT) to compute the electronic circular dichroism (ECD) spectra of the chiral compounds of urate degradation (HIU, OHCU, allantoin) and we compared the results with experimentally measured ECD spectra. The calculated ECD spectra for (S)-HIU and (S)-OHCU reproduced well the experimental spectra obtained through the enzymatic degradation of urate. Less conclusive results were obtained with allantoin, although the computed optical rotations in the transparent region supported the original assignment of the (+)-S configuration. These absolute configuration assignments can facilitate the study of the enzymes involved in urate metabolism and help us to understand the mechanism leading to the toxicity of urate oxidation products. PMID- 21647521 TI - Chiral indane skeleton based thiourea catalyzed highly stereoselective cascade Michael-enolation-cyclization reaction. AB - An efficient asymmetric cascade reaction catalyzed by a chiral bifunctional indane amine-thiourea catalyst has been developed. From a broad substrate scope, chiral dihydro-2H-pyran complexes that contained two stereogenic centers were obtained in a one-pot manner in good to excellent yields (72-97%) and high to excellent stereoselectivities (92-97% ee). PMID- 21647522 TI - Synthesis and characterization of [PW11O39Ir(H2O)]4-: successful incorporation of Ir into polyoxometalate framework and study of the substitutional lability at the Ir(III) site. AB - The first Keggin-type heteropolyanion, [PW(11)O(39)Ir(H(2)O)](4-) (1), was synthesized by hydrothermal reaction from two different polytungstate precursors and [IrF(6)](2-). It was isolated as (Bu(4)N)(4)[PW(11)O(39)Ir(H(2)O)] (1a), which was completely characterized by multinuclear (31)P and (183)W NMR, ESI-mass spectrometry and cyclic voltammetry. A rapid screening methodology to ascertain the intrinsic substitutional lability at the Ir site is also presented, based on ESI-MS. PMID- 21647523 TI - Differential interference contrast polarization anisotropy for tracking rotational dynamics of gold nanorods. AB - We describe differential interference contrast (DIC) polarization anisotropy for tracking rotational dynamics of gold nanorod (AuNR) probes. DIC polarization anisotropy enabled us to reveal the unidirectional clockwise circular translocation of an AuNR attached to a kinesin-driven microtubule and to precisely determine the real-time orientation of the AuNR during the dynamic process. PMID- 21647524 TI - Vertically aligned mixed V2O5-TiO2 nanotube arrays for supercapacitor applications. AB - Highly ordered mixed V(2)O(5)-TiO(2) nanotubes can be formed by self-organizing anodization of Ti-V alloys with vanadium content of up to 18 at%. In the resulting oxide nanotube arrays, the vanadium is electrochemically switchable leading to a specific capacitance that can reach up to 220 F g(-1) and an energy density of 19.56 Wh kg(-1) with perfect reversibility and long-term stability. Thus these mixed oxide nanotubes may be considered as a promising candidate for supercapacitors. PMID- 21647525 TI - beta-Cyclodextrin conjugated magnetic nanoparticles for diazepam removal from blood. AB - beta-CD conjugated magnetic nanoparticles that serve as a hemoadsorbent for diazepam removal are fabricated. The diazepam is arrested by the conjugated beta CD and then the adsorbed diazepam is efficiently removed by an external magnetic field. These particles have potential applications in hemoperfusion or separation of other toxins and drugs. PMID- 21647526 TI - A novel (3,36)-connected and self-interpenetrated metal-organic framework with high thermal stability and gas-sorption capabilities. AB - By using a ligand containing pyridyl and carboxylate groups as units, a novel (3,36)-connected and self-interpenetrated metal-organic framework was constructed, which exhibited high thermal stability and gas sorption capabilities. PMID- 21647527 TI - Microwave-assisted polyol synthesis of copper nanocrystals without using additional protective agents. AB - We report the synthesis of 2 nm copper nanocrystals (Cu NCs) via a microwave assisted polyol method without using additional protective and reducing agents. The Cu NCs are oxidation resistant and exhibit photoluminescence and highly stable properties in a colloidal dispersion. PMID- 21647528 TI - Insulating diamond particles as substrate for Pd electrocatalysts. AB - The suitability of insulating highly crystalline diamond particles as support for Pd based electrocatalysts is explored for the first time by evaluating the electrochemical stripping of CO and oxidation of formic acid in acid solutions. PMID- 21647529 TI - 9-Aryl-1,2-dihydropyrrolo[3,4-b]indolizin-3-one (Seoul-Fluor) as a smart platform for colorful ratiometric fluorescent pH sensors. AB - In this communication, we report that 9-aryl-1,2-dihydropyrrolo[3,4-b]indolizin-3 one (Seoul-Fluor) can serve as a potential platform for colorful ratiometric fluorescent pH sensors by simple incorporation of pH responsive elements on Seoul Fluor. Seoul-Fluor-based fluorescent pH sensors allow the emission- and pH-tuning ability upon protonation by varying their pK(a) values and electronic characteristics of substituents by a rational design. PMID- 21647530 TI - Guidelines for visualizing and annotating rule-based models. AB - Rule-based modeling provides a means to represent cell signaling systems in a way that captures site-specific details of molecular interactions. For rule-based models to be more widely understood and (re)used, conventions for model visualization and annotation are needed. We have developed the concepts of an extended contact map and a model guide for illustrating and annotating rule-based models. An extended contact map represents the scope of a model by providing an illustration of each molecule, molecular component, direct physical interaction, post-translational modification, and enzyme-substrate relationship considered in a model. A map can also illustrate allosteric effects, structural relationships among molecular components, and compartmental locations of molecules. A model guide associates elements of a contact map with annotation and elements of an underlying model, which may be fully or partially specified. A guide can also serve to document the biological knowledge upon which a model is based. We provide examples of a map and guide for a published rule-based model that characterizes early events in IgE receptor (FcepsilonRI) signaling. We also provide examples of how to visualize a variety of processes that are common in cell signaling systems but not considered in the example model, such as ubiquitination. An extended contact map and an associated guide can document knowledge of a cell signaling system in a form that is visual as well as executable. As a tool for model annotation, a map and guide can communicate the content of a model clearly and with precision, even for large models. PMID- 21647531 TI - Using logic programming for modeling the one-carbon metabolism network to study the impact of folate deficiency on methylation processes. AB - Dynamical modeling is an accurate tool for describing the dynamic regulation of one-carbon metabolism (1CM) with emphasis on the alteration of DNA methylation and/or dUMP methylation into dTMP. Using logic programming we present a comprehensive and adaptative mathematical model to study the impact of folate deficiency, including folate transport and enzymes activities. 5 Methyltetrahydrofolate (5mTHF) uptake and DNA and dUMP methylation were studied by simulating nutritional 5mTHF deficiency and methylenetetrahydrofolate reductase (MTHFR) gene defects. Both conditions had distinct effects on 1CM metabolite synthesis. Simulating severe 5mTHF deficiency (25% of normal levels) modulated 11 metabolites. However, simulating a severe decrease in MTHFR activity (25% of normal activity) modulated another set of metabolites. Two oscillations of varying amplitude were observed at the steady state for DNA methylation with severe 5mTHF deficiency, and the dUMP/dTMP ratio reached a steady state after 2 h, compared to 2.5 h for 100% 5mTHF. MTHFR activity with 25% of V(max) resulted in an increased methylated DNA pool after half an hour. We observed a deviation earlier in the profile compared to 50% and 100% V(max). For dUMP methylation, the highest level was observed with 25%, suggesting a low rate of dUMP methylation into dTMP with 25% of MTHFR activity. In conclusion, using logic programming we were able to construct the 1CM for analyzing the dynamic system behavior. This model may be used to refine biological interpretations of data or as a tool that can provide new hypotheses for pathogenesis. PMID- 21647532 TI - Fishing for function: zebrafish BAC transgenics for functional genomics. AB - Transgenics using bacterial artificial chromosomes (BACs) offers a great opportunity to look at gene regulation in a developing embryo. The modified BAC containing a reporter inserted just before the translational start site of the gene of interest allows for the visualization of spatio-temporal gene expression. Though this method has been used in the mouse model extensively, its utility in zebrafish studies is relatively new. This review aims to look at the utility of making BAC transgenics in zebrafish and its applications in functional genomics. We look at the various methods to modify the BAC, some limitations and what the future holds. PMID- 21647533 TI - Can recombinant thrombomodulin play a preventive role for veno-occlusive disease after haematopoietic stem cell transplantation? PMID- 21647534 TI - Molecular basis of hereditary factor V deficiency in India: identification of four novel mutations and their genotype-phenotype correlation. PMID- 21647535 TI - Evaluation of the PC-1 K121Q and G2906C variants as independent risk factors for ischaemic stroke. AB - Overexpression of plasma cell membrane glycoprotein-1 (PC-1) inhibits insulin receptor tyrosine kinase activity and thus favours insulin resistance and atherosclerotic vascular disease. Recent findings indicate that the minor variant K121Q in the PC-1 gene confers an increased risk for early myocardial infarction independent of other established risk factors. We hypothesized that genetic variants in PC-1 may also influence the risk for cerebrovascular disease. AIM: Therefore, we assessed the association of the PC-1 K121Q variant in the coding region and a polymorphism (G2906C) in the 3' untranslated region of the PC-1 gene with the risk of stroke. PATIENTS: We analyzed 1014 patients with a history of ischaemic stroke from the Vienna stroke registry and 1001 control individuals without vascular disease. RESULTS, CONCLUSION: Genotype frequencies of both genetic variants were similar in patients and controls in the total study population. By multivariate analysis, no interactions were observed between the PC-1 genotype and established vascular risk factors. However, the PC-1 2906C allele was significantly more frequent in patients who suffered from stroke before the age of 40 years. In these patients the risk for ischaemic stroke was increased four-fold. PMID- 21647536 TI - Small-molecule activators of AMP-activated protein kinase (AMPK), RSVA314 and RSVA405, inhibit adipogenesis. AB - AMP-activated protein kinase (AMPK) is a sensor and regulator of cellular energy metabolism potentially implicated in a broad range of conditions, including obesity and Alzheimer's disease. Its role in the control of key metabolic enzymes makes this kinase a central player in glucose and lipid homeostasis. Recently, by screening a library of synthetic small molecules selected for their structural similarity with the natural polyphenol resveratrol, we identified RSVA314 and RSVA405 as potent indirect activators of AMPK (half-maximal effective concentration [EC50] = 1 MUmol/L in cell-based assays). Here we show that RSVA314 and RSVA405 can significantly activate AMPK and inhibit acetyl-CoA carboxylase (ACC), one target of AMPK and a key regulator of fatty acid biogenesis, in nondifferentiated and proliferating 3T3-L1 adipocytes. We found that RSVA314 and RSVA405 treatments inhibited 3T3-L1 adipocyte differentiation by interfering with mitotic clonal expansion during preadipocyte proliferation (half-maximal inhibitory concentration [IC50] = 0.5 MUmol/L). RSVA314 and RSVA405 prevented the adipogenesis-dependent transcriptional changes of multiple gene products involved in the adipogenic process, including peroxisome proliferator-activated receptor (PPAR)-gamma, CCAAT/enhancer-binding protein alpha (C/EBPalpha), fatty acid synthase, fatty acid binding protein 4 (aP2), RANTES or resistin. Furthermore, orally administered RSVA405 at 20 and 100 mg/kg/d significantly reduced the body weight gain of mice fed a high-fat diet. This work shows that the novel small molecule activators of AMPK (RSVA314 and RSVA405) are potent inhibitors of adipogenesis and thus may have therapeutic potential against obesity. PMID- 21647538 TI - Environmental drug prevention in the EU. Why is it so unpopular? AB - Adolescents go through changes in their neurobehavioural and psychosocial functioning that can result in their desire to conform to peer norms taking predominance over cognitive impulse control mechanisms. This can help explain why, when peers are present, adolescent behaviour may not be significantly modified by what they know about risks and consequences. This finding has implications for prevention as it points to the importance of responses that actively target the environment in which substance use takes place, and where social norms are formed and supported. In practice however, across Europe information-only prevention approaches - despite a weak evidence for their effectiveness - continue to be most commonly used. Why this is the case is the question addressed by this editorial. A definition for environmental prevention is also provided: as strategies that aim to alter physical, social and economic environment without relying on persuasion. Boundaries of the definition with health promotion are discussed and the available evidence for the efficacy of this perspective reviewed, as is the information on the availability of environmental prevention within Europe. These data do not support the contention made by some member states that their prevention strategies are comprehensive and cover all addictive substances. Overall, although environmental approaches are becoming more common, they are disproportionately found in the North of Europe, and are most noticeable in three areas: at the macro-level in tobacco bans and alcohol policies, in strategies to improve the school environment and ethos, and in local level policies to regulate recreational settings. PMID- 21647537 TI - Effect of oxygen levels on the physiology of dendritic cells: implications for adoptive cell therapy. AB - Dendritic cell (DC)-based adoptive tumor immunotherapy approaches have shown promising results, but the incidence of tumor regression is low and there is an evident call for identifying culture conditions that produce DCs with a more potent Th1 potential. Routinely, DCs are differentiated in CO(2) incubators under atmospheric oxygen conditions (21% O(2)), which differ from physiological oxygen levels of only 3-5% in tissue, where most DCs reside. We investigated whether differentiation and maturation of DCs under physiological oxygen levels could produce more potent T-cell stimulatory DCs for use in adoptive immunotherapy. We found that immature DCs differentiated under physiological oxygen levels showed a small but significant reduction in their endocytic capacity. The different oxygen levels did not influence their stimuli-induced upregulation of cluster of differentiation 54 (CD54), CD40, CD83, CD86, C-C chemokine receptor type 7 (CCR7), C-X-C chemokine receptor type 4 (CXCR4) and human leukocyte antigen (HLA) DR or the secretion of interleukin (IL)-6, tumor necrosis factor (TNF)-alpha and IL-10 in response to lipopolysaccharide (LPS) or a cytokine cocktail. However, DCs differentiated under physiological oxygen level secreted higher levels of IL 12(p70) after exposure to LPS or CD40 ligand. Immature DCs differentiated at physiological oxygen levels caused increased T-cell proliferation, but no differences were observed for mature DCs with regard to T-cell activation. In conclusion, we show that although DCs generated under atmospheric or physiological oxygen conditions are mostly similar in function and phenotype, DCs differentiated under physiological oxygen secrete larger amounts of IL-12(p70). This result could have implications for the use of ex vivo-generated DCs for clinical studies, since DCs differentiated at physiological oxygen could induce increased Th1 responses in vivo. PMID- 21647539 TI - Reaction to the article: environmental drug prevention in the EU - why is it so unpopular? by Gregor Burkhart. PMID- 21647540 TI - Quality of illegally and informally produced alcohol in Europe: Results from the AMPHORA project. AB - BACKGROUND: In the WHO region Europe, the average unrecorded adult per capita alcohol consumption was 2.67 L pure ethanol in 2005, which is 22% of the total consumption of 12.20 L. Despite concerns about potential health harms from the chemical composition of unrecorded alcohol, there are surprisingly few data on the problem in the European Region. This study reports the results from the Alcohol Measures for Public Health Research Alliance (AMPHORA) project, which assessed the quality of unrecorded alcohol in a Europe-wide study. METHODS: Samples of unrecorded alcohol were collected in 16 European countries and chemically analyzed for potentially health-relevant parameters. Thresholds for parameters were defined based on potential health hazards of daily drinking. RESULTS: The average alcoholic strength of unrecorded wine products was 14.9% vol, and 47.8% vol in unrecorded spirits. One half of the samples (n=57) showed acceptable alcohol quality. The other half (n=58) showed one or several deficits with the most prevalent problem being ethyl carbamate contamination (n=29). Other problems included copper (n=20), manganese (n=16) and acetaldehyde (n=12). All other parameters (including methanol, higher alcohols, phthalates) were only seldom problematic (limit exceedance in less than 10 samples). The price of unrecorded alcohol was approximately 45% of the price of recorded alcohol. CONCLUSIONS: The major problem regarding unrecorded alcohol appears to be ethanol itself, as it is often higher in strength and its lower price may further contribute to higher drinking amounts. Compared to the health effects of ethanol, the contamination problems detected may be of minor importance as exposure will only in worst-case scenarios reach tolerable daily intakes of these substances. PMID- 21647541 TI - [Nicotine dependence and readiness to quit smoking in the Spanish population]. AB - OBJECTIVE: To describe the nicotine dependence and readiness to quit smoking in the smoker population. METHODS: Cross-sectional study on a representative sample of the Spanish population of >=18 years old. We gathered information by means of telephone interviews conducted between June and July of 2006. We studied the nicotine dependence with the Fagerstrom Test for Nicotine Dependence (FTND) and the readiness to quit according to the stages of change from the Transtheoretical Model in a sample of cigarette smokers. RESULTS: 22.5% of participants (95% CI: 20.9-24.2%) smokers cigarettes. They smoked an average of 14.4 cigarettes per day (standard deviation 9.15) and the mean FTND score was 2.8, with no differences by the stages of change. 64.3% (95% CI: 60.3-68.2%) of smokers were in the precontemplation stage, 25.4% (95% CI: 21.8-28.9%) in contemplation, and 10.4% (95% CI: 7.9-12.9%) in preparation, with no differences by sex. The most nicotine dependent smokers (FTND>=6) had mainly primary studies, started to smoke at earlier ages, and smoked more cigarettes per day. CONCLUSIONS: Most Spanish smokers have low nicotine dependence and are in precontemplation stage. Smoking cessation programmes should be addressed to reduce dependence, help smokers to progress through the stages of change, and, consequently, reduce the prevalence of smokers in the population. PMID- 21647542 TI - [Neuropsychological aspects of nicotine craving]. AB - Craving has been defined as the motivation to self-administer a substance previously consumed. It has been hypothesized that craving contributes significantly to compulsive drug use and relapse after a period of abstinence in humans. Neuropsychological and brain-imaging studies have identified numerous brain regions that may be involved in craving. In this paper, the neuropsychological mechanisms of craving for nicotine are reviewed, focusing on three systems that appear to be involved in craving states. First of all, the reward system, responsible for the development of dependence and craving. Secondly, the emotional and associative system, which is related to conditioned craving. And third, the system involved in the neural basis of cognitive and decision making processes. The most influential theoretical models on craving are also reviewed, including those based on conditioning mechanisms, on cognitive mechanisms and on cognitive-behavioral mechanisms, as well as the neurobiological model. Factors related to the evaluation and treatment of craving are also discussed, with particular emphasis on clinical aspects. Finally, we stress the importance of a multidisciplinary approach for achieving a common model on craving and improving the diagnostic tools and treatment strategies. PMID- 21647543 TI - [Relation between personality traits and personal values in cocaine-dependent patients]. AB - OBJECTIVE: To describe the relationship between personal values and personality traits in cocaine-using patients and analyze their specificity in the explanation of different types of constructs. METHOD: A study was carried out to explore the association between these variables in a group of 230 patients receiving treatment for cocaine dependence. The Portrait Values Questionnaire was used for measuring personal values, while the Big-Five Factors Questionnaire was used to measure personality traits. In addition, we explored the relationship of values and traits with the variables "degree of satisfaction with life" (life satisfaction) and "belonging to a religious association" (religiosity). RESULTS: A significant association was found between personal values and personality traits. At the same time, their conceptual and empirical differences were revealed, as it was demonstrated that personal values better explain "belonging to a religious association", whilst personality traits better explain "degree of satisfaction with life". Thus, it was found that personal values better explain behaviours that depend on greater cognitive control, while personality traits would have more influence on tendencies and behaviours that are subject to lower cognitive control levels. CONCLUSIONS: Considering the relationship between the two constructs, and given that cocaine use is associated with both high and low cognitive control, in explanations of cocaine use it would be appropriate to take into account the explanatory contribution of personal values and personality traits in a complementary way. PMID- 21647544 TI - [Addiction, impulsivity and temporal curves of desire]. AB - This study addresses, from theories of associative learning, the impulsivity and craving in patients with a diagnosis of substance dependence in abstinence, habitual use and relapse. We present a new method for the assessment of desire based on discount curve self description, in which patients describe how they perceive the temporal evolution and intensity of their desire. At the same time, through a brief questionnaire about emotion and desire -developed by the authors and the Plutchik Impulsivity Scale, we set out to check whether positive or negative emotions increase, decrease, or are uninvolved in changes of desire and impulsivity. Perception of the evolution of desire in patients from the Relapse and Habitual Use groups is similar and presents a hyperbolic curve, but the Relapse group presented greater impulsivity. Moreover, a relationship is found in patients from the Relapse group between the hyperbolic increase of desire, the pleasurable states and impulsivity. The higher scores on impulsivity and the hyperbolic evolution of desire in patients who have recently suffered a relapse indicate the need to consider and specifically address these aspects as relevant to psychotherapy and/or pharmacological therapy. PMID- 21647545 TI - [Amisulpride for the treatment of alcohol dependence]. AB - BACKGROUND: 6-month naturalistic, open-label trial to compare amisulpride versus topiramate and naltrexone as a treatment for patients with alcohol dependence, with assessments at enrolment and after 3 and 6 months of treatment. METHODS: 274 alcohol-dependent patients who had been drinking heavily during the past month were included. Once detoxified, patients were assigned to one of three treatment groups (naltrexone 50 mgr per day, topiramate 200 mgr per day or amisulpride 100 mgr per day). Patients were assessed at baseline and after 3 and 6 months of follow-up. Outcome was measured using tools that assessed alcohol intake (EuropASI and Alcohol Timeline Followback), craving (OCDS), disability (WHO/DAS), and quality of life (EQ-5D); changes in biomarkers of alcohol intake were also noted. RESULTS: at the 6-month follow-up patients taking amisulpride had poorer results than those taking topiramate in direct measures of alcohol intake (OCDS, alcohol intake, number of drinks per day and heavy drinking days), but no significant differences were found in these measures on comparing the amisulpride patients with those taking naltrexone. CONCLUSIONS: in this study, amisulpride, at a dose of 100 mgr per day, was less effective than topiramate, at a dose of 200 mg per day, but as effective as naltrexone, at a dose of 50 mg per day, for reducing alcohol intake and craving over the period of the study. PMID- 21647546 TI - [Multidimensional alcohol craving scale and [123I] iodobenzamide SPECT as predictors of early relapse in alcohol-dependent patients]. AB - BACKGROUND: The Multidimensional Alcohol Craving Scale (MACS) and Single Photon Emission Computerized Tomography (SPECT) with (123)I-iodobenzamide ((123)I-IBZM) can be useful tools for assessing relapse risk in early recovery from alcohol dependency. The aim of this study was to assess possible relationships between MACS score, (123)I-IBZM binding and time to first heavy drinking day (TFHD) after detoxification treatment. METHODS: Nineteen alcohol-dependent in-patients were evaluated by MACS scale and an 123I-IBZM-SPECT, performed following alcohol detoxification treatment. At discharge, participants were advised to take naltrexone 50 mg/day for relapse prevention. TFHD was assessed over a 12-week follow up. RESULTS: The MACS score at the beginning of the detoxification process and naltrexone treatment after detoxification were independent predictive factors for TFHD. CONCLUSIONS: The MACS scale is a better predictor of TFHD than IBZM binding. It is simple, non-invasive and inexpensive and appears to be a useful instrument both for clinical practice and for research. PMID- 21647547 TI - [Epidemiological and diagnostic axis I gender differences in dual diagnosis patients]. AB - INTRODUCTION: Dual diagnosis is the co-occurrence of a substance abuse disorder and a psychiatric condition. Gender has been found to be associated with differences in prevalence of mental disorders as well as outcome, prognosis and treatment-seeking. MATERIAL AND METHODS: Articles published in Medline, Web of Science and Journal Citation Reports up to December 2009 that examined gender, prevalence and clinical characteristics of dual-diagnosis patients aged over 18 were reviewed. CONCLUSIONS: The distribution of Axis I disorders by gender is similar for dually diagnosed patients and single-disorder patients. The prevalence of psychotic and bipolar disorders is higher in men, whereas anxiety and affective disorders are more prevalent in women. Dually diagnosed females with psychotic disorders do not show better prognosis than men. Finally, polydrug use among dual-diagnosis individuals is more prevalent in males. PMID- 21647548 TI - Distribution and expression characteristics of triterpenoids and OSC genes in white birch (Betula platyphylla suk.). AB - Betulin and oleanolic acids (pentacyclic triterpenoid secondary metabolites) have broad pharmacological activities and can be potentially used for the development of anti-cancer and anti-AIDS drugs. In this study, we detected the accumulation and the distribution characteristics of betulin and oleanolic acid in various organs of white birch at different ages. We also determined the expression of 4 OSC genes (LUS, beta-AS, CAS1 and CAS2) involved in the triterpenoid synthesis pathways by real time RT-PCR. The result showed that the 1-year old birch can synthesize betulin and oleanolic acid. In addition, betulin and oleanolic acids were mainly distributed in the bark, while the content in the root skin and leaf was very low. The content of betulin and oleanolic acid in birch varied in different seasons. The content of betulin and oleanolic acid and their corresponding LUS and beta-AS gene expression were very low in 1-year old birch. With increasing age of birch, betulin content was increased, while oleanolic acid was decreased. Similar changes were also observed for their corresponding synthesis genes LUS and beta-AS. In the leaf of 1-year old plant, the highest expression of CAS1 and CAS2 occurred at end of September, while expression of LUS and the beta-AS was low from June to October. In the stem skin,high expression of beta-AS and the LUS genes occurred from the end of July to September. In the root, high expression of the beta-AS gene was observed at the end of October. These results indicated that triterpenoid gene expression was similar to the triterpene accumulation. Expression of LUS gene and beta-AS gene in birch with different ages were corresponding to the betulinic and oleanolic acid accumulation. Expression of CAS1 and CAS2 genes were elevated with increasing age of birch. This study provides molecular mechanisms of triterpenes synthesis in birch plants. PMID- 21647549 TI - Molecular cloning, characterization and expression of a chalcone reductase gene from Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao. AB - A chalcone reductase (CHR) gene was isolated from Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao (A. mongholicus). The full-length cDNA of A. mongholicus CHR, designated as Amchr (GenBank accession No. HM357239), was 1196 bp long. It had a 957 bp open reading frame encoding a 318-amino acid protein of 35 kDa, a 67 bp 5' non-coding region and a 172 bp 3'-untranslated region. The putative AmCHR protein showed striking similarity to CHR from other leguminous species. Two-dimensional structure modeling showed that AmCHR consisted of 45.28% alpha-helix, 10.38% extended strand and 44.34% random coil. Prediction showed that three-dimensional AmCHR was a global protein containing an aldo-ket-red domain, with a putative Asp-Tyr-Lys-His catalytic tetrad in the center. The AmCHR gene was 1251 bp long, consisting of three exons and two introns. Intron I was 125 bp and intron II was 169 bp long. Southern blot analysis indicated that Amchr belonged to a small multigene family. Under natural conditions, Amchr was expressed differentially in the root, stem and leaf tissues of A. mongholicus, with a preferential expression in the root. The recombinant AmCHR protein was successfully expressed in Escherichia coli strain BL21 with pET42a vector. The result showed that the expressed AmCHR protein had molecular weight of about 35 kDa, which matched the size of the predicted protein by bioinformatic analysis. This study opened avenues towards understanding of the function of AmCHR protein and the role of the Amchr gene in the calycosin-7-O-beta-D: -glucoside branch pathway in A. mongholicus. PMID- 21647550 TI - An association between polymorphism of the heme oxygenase-1 and -2 genes and age related macular degeneration. AB - Iron may be implicated in the generation of oxidative stress by the catalyzing the Haber-Weiss or Fenton reaction. On the other hand, oxidative stress has been implicated in the pathogenesis of age-related macular degeneration (AMD) and heme oxygenase-1 (HO-1), encoded by the HMOX1 gene and heme oxygenase-2 (HO-2), encoded by the HMOX2 gene are important markers of iron-related oxidative stress and its consequences. Therefore, variability of the HMOX1 and HMOX2 genes might be implicated in the pathogenesis of AMD through the modulation of the cellular reaction to oxidative stress. In the present work, we investigated the association between AMD and a G -> C transversion at the 19 position in the HMOX1 gene (the 19G>C-HMOX1 polymorphism, rs2071747) and a A -> G transition at the -42 + 1444 position in the HMOX2 gene (the -42 + 1444A>G-HMOX2 polymorphism, rs2270363) and its modulation by some environmental factors. 279 patients with AMD and 105 controls were recruited in this study and the polymorphisms were typed by restriction fragment length polymorphism and allele-specific polymerase chain reaction (PCR). We observed an association between the occurrence of dry AMD and the G/A genotype of the -42 + 1444A>G-HMOX2 polymorphism (odds ratio (OR) 2.72), whereas the G/G genotype reduced the risk of dry AMD (OR 0.41). The G/C genotype and the C allele of the 19 G>C-HMOX1 polymorphism and the G/G genotype and the G allele of the -42 + 1444A>G-HMOX2 polymorphism were associated with progression of AMD from dry to wet form (OR 4.83, 5.20, 2.55, 1.69, respectively). On the other hand, the G/G genotype and the G allele of the 19 G>C HMOX1 polymorphism and the A/G genotype and the A allele of the -42 + 1444A>G HMOX2 polymorphism protected against AMD progression (OR 0.19, 0.19, 0.34, 0.59, respectively). Therefore, the 19G>C-HMOX1 and the -42 + 1444A>G-HMOX2 polymorphisms may be associated with the occurrence and progression of AMD. PMID- 21647551 TI - The expression of HIF-1alpha in primary hepatocellular carcinoma and its correlation with radiotherapy response and clinical outcome. AB - The purpose of this study was to evaluate the relationship between hypoxia inducible factor-1alpha (HIF-1alpha) protein expression in hepatocellular carcinoma (HCC), and responses of abdominal metastatic lymph nodes (LNs) from HCC patients treated with external beam radiotherapy (EBRT). HIF-1alpha immunohistochemical staining was performed on tissue microarrays (TMAs) of primary HCC specimens from 69 HCC patients with abdominal LN metastases. All patients received abdominal metastatic LN EBRT at the Department of Radiation Oncology at Zhongshan Hospital. A receiver-operating characteristic (ROC)-based approach and logistical regression analysis were used to determine the predictive value of HIF-1alpha expression in primary tumors with HCC metastatic LN EBRT response. Kaplan-Meier curves and log-rank tests were used to analyze patient survival. Cox proportional hazards regression model was used to analyze independent prognostic factors. HIF-1alpha expression was correlated with blood hemoglobin (Hb: r = -0.280, P = 0.020), response of abdominal metastatic LNs to EBRT (r = 0.286, P = 0.017), locoregional recurrence (r = 0.278, P = 0.021), and cancer-specific deaths (r = 0.298, P = 0.013). HIF-1alpha expression was predictive of EBRT response of metastatic LNs [area under the curve (AUC): 0.646; 95% confidence interval (CI): 0.499-0.793; P = 0.047], locoregional recurrence (AUC: 0.657; 95% CI: 0.509-0.805; P = 0.049) and cancer-specific deaths (AUC: 0.671; 95% CI: 0.531-0.812; P = 0.035). Patients with tumors exhibiting high HIF 1alpha expression had significantly poorer overall survival (OS) than those with low tumor expression of HIF-1alpha (P = 0.016). Multivariate analysis showed that Hb (P = 0.035), vascular invasion (P = 0.026), Child-Pugh score (P < 0.001), intrahepatic tumor control (P < 0.001), and HIF-1alpha (P = 0.020) were independent prognosis factors for OS of HCC patients after receiving abdominal metastatic LN EBRT. HIF-1alpha expression in primary HCCs was associated with EBRT response of abdominal metastatic LNs and poor prognosis. PMID- 21647552 TI - Stem cell course in the Middle East: science diplomacy and international collaborations during the Arab spring. AB - In April 2011, an international advanced course and workshop entitled "Frontiers in Human Pluripotent Stem Cells" and an International Congress on Fertility and Genetics ( http://www.fertigen.com.jo/ConferenceDetails.aspx ) was held in Amman Jordan hosted by the Jordanian Society of Fertility and Genetics under the auspices of the International Cell Research Organization (ICRO), a UNESCO associated NGO. The Congress President Dr. Zaid Kilani, with Dr. Abdel Latif Abu Khadra, President of the Jordanian society for Fertility and Genetics, Dr. Rana Dajani of the Hashemite University of Jordan, and their Organizing Committee proved to be an excellent organizers and dedicated physician-scientists and, focusing on fertility, genetics and stem cells in a wide range of advanced therapeutic applications. Brilliant course participants included trainees, scientists and clinicians from the Greater Middle East. The lectures and practical sessions, presented by internationally acknowledged scientists, included overviews of recent achievements in pluripotent stem cell research, emphasizing the role of both the embryonic (ES) and induced pluripotent stem (iPS) cells. A major emphasis was placed on the clinical achievements in germ cell and umbilical cord stem cell transplantation issues, and on the potential of fast and successful prenatal and pre-implantation molecular genetics diagnostics. The organization of the stem cell course in the Holy Land especially emphasized that issues of "eternal life" and "rejuvenation" are already at hand--at least in the pluripotent stem cell research field. In the lively atmosphere of the course about 60 participants had heated discussions on the possibility and ethics of advanced prenatal diagnostics, and on regulatory issues reflecting the need of separation of clinically effective versus unapproved, unwarranted stem cell treatments. An open discussion of many ethical issues, reflecting profound differences in religion and medical tradition in the different countries, made this course exceptionally interesting for both teachers and trainees. PMID- 21647553 TI - Mental health matters in elementary school: first-grade screening predicts fourth grade achievement test scores. AB - The objective of the study was to evaluate whether mental health problems identified through screens administered in first grade are related to poorer academic achievement test scores in the fourth grade. The government of Chile uses brief teacher- and parent-completed measures [Teacher Observation of Classroom Adaptation-Revised (TOCA-RR) and Pediatric Symptom Checklist (PSC-Cl)] to screen for mental health problems in about one-fifth of the country's elementary schools. In fourth grade, students take the national achievement tests (SIMCE) of language, mathematics and science. This study examined whether mental health problems identified through either or both screens predicted achievement test scores after controlling for student and family risk factors. A total of 17,252 students had complete first grade teacher forms and these were matched with fourth grade SIMCE data for 11,185 students, 7,903 of whom also had complete parent form data from the first grade. Students at risk on either the TOCA-RR or the PSC-Cl or both performed significantly worse on all SIMCE subtests. Even after controlling for covariates and adjusting for missing data, students with mental health problems on one screen in first grade had fourth grade achievement scores that were 14-18 points (~1/3 SD) lower than students screened as not at risk. Students at risk on both screens had scores that were on average 33 points lower than students at risk on either screen. Mental health problems in first grade were one of the strongest predictors of lower achievement test scores 3 years later, supporting the premise that for children mental health matters in the real world. PMID- 21647554 TI - Supporting researchers: mentoring and awards of the Journal. PMID- 21647555 TI - Vitamin D in HIV-Infected Patients. AB - Observational studies have noted very high rates of low 25(OH)D (vitamin D) levels in both the general and HIV-infected populations. In HIV-infected patients, low 25(OH)D levels are likely a combination of both traditional risk factors and HIV-specific and antiretroviral therapy-specific contributors. Because of this unique risk profile, HIV-infected persons may be at greater risk for low 25(OH)D levels and frank deficiency and/or may respond to standard repletion regimens differently than HIV-uninfected patients. Currently, the optimal repletion and maintenance dosing regimens for HIV-infected patients remain unknown, as do potential benefits of supplementation that may be unique to the HIV-infected population. This paper reviews data published on HIV infection and vitamin D health in adults over the last year. PMID- 21647556 TI - Multiple phenotypes in genome-wide genetic mapping studies. AB - For many psychiatric and other traits, diagnoses are based on a number of different criteria or phenotypes. Rather than carrying out genetic analyses on the final diagnosis, it has been suggested that relevant phenotypes should be analyzed directly. We provide an overview of statistical methods for the joint analysis of multiple phenotypes in case-control association studies. PMID- 21647558 TI - Comparison of axial force and cell width of self-expandable metallic stents: which type of stent is better suited for hilar biliary strictures? AB - BACKGROUND: Various types of self-expandable metallic stents (SEMSs) are commercially available. However, few reports have compared the performance of the various SEMSs. In addition to long-term patency, maneuverability during initial placement and feasibility of re-intervention for the stent occlusion are also very important. METHODS: In this retrospective analysis, we compared the duration of stent patency and frequency of re-interventions in 96 patients diagnosed with unresectable hilar biliary strictures in whom initial 10-mm SEMSs were inserted between June 1999 and November 2008. RESULTS: Based on Kaplan-Meier curves, significantly shorter patency duration was noted for SEMSs with moderate axial force (AF) than for other groups of SEMSs with low AF. Endoscopic re interventions for SEMS occlusion were easier in a group of SEMSs with low AF and large cell width than in other groups of SEMSs. CONCLUSIONS: With respect to patency duration and the frequency of re-intervention required, we consider that SEMSs with low AF and large cell width are favorable for hilar biliary strictures. PMID- 21647559 TI - Ischemic preconditioning prior to intermittent Pringle maneuver in liver resections. AB - BACKGROUND/PURPOSE: Continuous inflow vascular occlusion during liver resections causes less severe ischemia and reperfusion injury (IRI) if it is preceded by ischemic preconditioning (IP) or if intermittent inflow occlusion is used during the resection. No previous clinical trial has studied the effects of adding IP to intermittent inflow occlusion. METHODS: Consecutive patients (n = 32) with suspicion of malignant liver disease had liver resections (minimum 2 segments) performed with inflow occlusion (intermittent clamping in a manner of 15 min of ischemia and 5 min of reperfusion repetitively; 15/5). Half of the patients were randomized to receive IP (10 min of ischemia and 10 min of reperfusion before parenchymal transection; 10/10). The patients were stratified according to volume of resection and none had chronic liver disease. The patients were followed for 5 days with microdialysis (MUD). RESULTS: All patients completed the study and there were no deaths. No differences were seen between the groups regarding demographics or perioperative parameters (bleeding, duration of ischemia, resection volume, complications, and serum laboratory tests). There were no differences in alanine aminotransferase (ALT), aspartate aminotransferase (AST), bilirubin, or prothrombin time (PT)-INR levels, but MUD revealed lower levels of lactate, pyruvate, and glucose in the IP group having major liver resections (analysis of variance; ANOVA). Nitrite and nitrate levels in MUD decreased postoperatively, but no differences were seen between the groups. In one patient an elevated MUD-glycerol curve was seen before the diagnosis of a stroke was made. CONCLUSIONS: IP before intermittent vascular occlusion does not reduce the serum parameters used to assess IRI. IP seems to improve aerobic glucose metabolism, as the levels of glucose, pyruvate, and lactate locally in the liver were reduced, compared to controls, in patients having >3 segments resected. MUD may be used to monitor metabolism locally. PMID- 21647560 TI - A phase II study of oral S-1 with concurrent radiotherapy followed by chemotherapy with S-1 alone for locally advanced pancreatic cancer. AB - BACKGROUND/PURPOSE: S-1 is a new oral fluoropyrimidine anticancer agent shown to be effective for pancreatic cancer. In a previous phase I trial, we evaluated the safety of S-1 combined with radiotherapy to determine the maximum tolerated dose and dose-limiting toxicity in patients with unresectable pancreatic cancer. The recommended dose of S-1 for phase II trials of chemoradiotherapy was determined as 80 mg/m(2)/day given on days 1-21 of a 28-day cycle. This phase II study was conducted to further evaluate the efficacy and toxicity of radiotherapy combined with S-1 (UMIN000004794). METHODS: Eligible patients had locally advanced and unresectable pancreatic cancer without distant metastases, an Eastern Cooperative Oncology Group performance status of 0-1, adequate organ and marrow functions, and no prior anticancer therapy. Patients initially received 4 weeks of chemoradiotherapy. S-1 was given orally at a dose of 80 mg/m(2)/day twice daily on days 1-21. Radiotherapy was delivered in fractions of 1.25 Gy twice daily, 5 days per week for 4 weeks (total dose: 50 Gy in 40 fractions). One month after the completion of chemoradiotherapy, S-1 was administered for 14 days followed by a 14-day rest period. This cycle was repeated as maintenance therapy until disease progression or unacceptable toxicity. RESULTS: Fifty patients were enrolled in this phase II study. Median follow-up was 14.6 months (range 5.4-58.9 months). Forty-three patients (86%) completed the scheduled course of chemoradiotherapy. There was no treatment-related death or grade 4 toxicity. The major toxic effects were leukopenia and nausea. The objective tumor response according to the Response Evaluation Criteria in Solid Tumours criteria was partial response in 15 patients (30%) (95% confidence interval (CI), 18-45%), stable disease in 23 (46%), and progressive disease in 12 (24%). Median progression-free survival and median overall survival were 6.7 months (95% CI, 4.7-11.2 months) and 14.3 months (95% CI, 10.8-20.8 months), respectively. Survival rates at 1 and 2 years were 62 and 27%, respectively. CONCLUSIONS: Combination therapy with S-1 and radiation in patients with locally advanced and unresectable pancreatic cancer is considered a promising, well-tolerated regimen that can be recommended as an effective treatment for locally advanced pancreatic cancer. PMID- 21647561 TI - Immunohistological markers for proliferative events, gliogenesis, and neurogenesis within the adult hippocampus. AB - Biologists long believed that, once development is completed, no new neurons are produced in the forebrain. However, as is now firmly established, new neurons can be produced at least in two specific forebrain areas: the subventricular zone (SVZ) and the dentate gyrus (DG) of the hippocampal formation. Neurogenesis within the adult DG occurs constitutively throughout postnatal life, and the rate of neurogenesis within the DG can be altered under various physiological and pathophysiological conditions. The process of adult neurogenesis within the DG is a multi-step process (proliferation, differentiation, migration, targeting, and synaptic integration) that ends with the formation of a post-mitotic functionally integrated new neuron. Various markers are expressed during specific stages of adult neurogenesis. The availability of such markers allows the time-course and fate of newly born cells to be followed within the DG in a detailed and precise fashion. Several of the available markers (e.g., PCNA, Ki-67, PH3, MCM2) are markers for proliferative events, whereas others are more specific for early phases of neurogenesis and gliogenesis within the adult DG (e.g., nestin, GFAP, Sox2, Pax6). In addition, markers are available allowing events to be distinguished that are related to later steps of gliogenesis (e.g., vimentin, BLBP, S100beta) or neurogenesis (e.g., NeuroD, PSA-NCAM, DCX). PMID- 21647562 TI - Vav1 is a crucial molecule in monocytic/macrophagic differentiation of myeloid leukemia-derived cells. AB - Vav1 is a critical signal transducer for both the development and function of normal hematopoietic cells, in which it regulates the acquisition of maturation related properties, including adhesion, motility, and phagocytosis. Vav1 is also important for the agonist-induced maturation of acute promyelocytic leukemia (APL)-derived promyelocytes, in which it promotes the acquisition of a mature phenotype by playing multiple functions at both cytoplasmic and nuclear levels. We investigated the possible role of Vav1 in the differentiation of leukemic precursors to monocytes/macrophages. Tumoral promyelocytes in which Vav1 was negatively modulated were induced to differentiate into monocytes/macrophages with phorbol-12-myristate-13-acetate (PMA) and monitored for their maturation related properties. We found that Vav1 was crucial for the phenotypical differentiation of tumoral myeloid precursors to monocytes/macrophages, in terms of CD11b expression, adhesion capability and cell morphology. Confocal analysis revealed that Vav1 may synergize with actin in modulating nuclear morphology of PMA-treated adherent cells. Our data indicate that, in tumoral promyelocytes, Vav1 is a component of lineage-specific transduction machineries that can be recruited by various differentiating agents. Since Vav1 plays a central role in the completion of the differentiation program of leukemic promyelocytes along diverse hematopoietic lineages, it can be considered a common target for developing new therapeutic strategies for the various subtypes of myeloid leukemias. PMID- 21647563 TI - Dynamics of expression of ARID1A and ARID1B subunits in mouse embryos and in cells during the cell cycle. AB - The mammalian SWI/SNF chromatin remodeling complexes play essential roles in cell cycle control through the transcriptional regulation of cell-cycle-specific genes. These complexes depend on the energy of ATP hydrolysis provided by the BRG1 or BRM catalytic subunit. They contain seven or more noncatalytic subunits, some being constitutive components, with others having paralogs that assemble in a combinatory manner producing different SWI/SNF-related complexes with specific functions. ARID1A and ARID1B are mutually exclusive subunits of the BAF complex. The specific presence of these subunits in the complex has been demonstrated to determine whether SWI/SNF functions as a corepressor (ARID1A) or as a coactivator (ARID1B) of the cell cycle genes. Our aim has been to analyze the relevance of the ARID1 subunits in development. We have compared the patterns of expression of these two genes through various mouse embryonic stages. Arid1a is expressed widely and intensively, whereas Arid1b is poorly transcribed and expressed in selected regions. Moreover, ARID1A and ARID1B present different kinetics of expression in the cell cycle. ARID1A accumulates in G0 and is downregulated throughout the cell cycle phases but is completely eliminated during mitosis, whereas ARID1B is expressed at comparable levels at all phases, even during mitosis. These kinetics probably affect the incorporation patterns of the ARID1 proteins to the complex and hence modulate SWI/SNF activity during proliferation and arrest. PMID- 21647564 TI - c-Ski in health and disease. AB - c-Ski is an evolutionary conserved protein that is involved in diverse cellular processes such as proliferation, differentiation, transformation, and tumor progression. A large range of cellular partners of c-Ski, including transcription factors, chromatin-remodeling molecules, tumor suppressors, and nuclear hormone receptors, has been identified. Moreover, numerous mechanisms have been described by which c-Ski regulates essential signaling pathways, e.g., the TGFbeta pathway. In this review, we summarize the diverse roles attributed to c-Ski during normal development and in cancer progression and discuss future strategies to unravel further the complex nature of c-Ski actions in a context-dependent manner. PMID- 21647565 TI - Bevacizumab for serous changes originating from a persistent branching vascular network following photodynamic therapy for polypoidal choroidal vasculopathy. AB - PURPOSE: To report the efficacy of intravitreal bevacizumab for either recurrent or persistent serous changes originating from a persistent branching vascular network or secondary choroidal neovascularization after photodynamic therapy for polypoidal choroidal vasculopathy, despite regression of polypoidal lesions. METHODS: Twenty eyes of 20 patients with at least 12 months of regular follow-up were retrospectively reviewed. Intravitreal bevacizumab was administered on an as needed basis, guided by optical coherence tomography (OCT), after the first injection. RESULTS: Seventeen (85%) of 20 eyes showed resolution of macular fluid after a mean of 1.9 (range 1-3) consecutive injections; however, 15 (88%) of them had relapsing episodes of fluid collection. The mean number of injections needed was 4.2 (range 1-6) over a period of 12 months. At 12 months, 10 eyes (50%) had no fluid accumulation on OCT, while 10 eyes (50%) had some residual fluid. The mean central foveal thickness improved significantly from 280 +/- 37 to 226 +/- 62 MUm (P = 0.002). Visual acuity was maintained or improved in 16 eyes (80%). CONCLUSIONS: Intravitreal bevacizumab appears to be effective in resolving intraretinal and subretinal fluid originating from these lesions. However, the favorable effect was maintained for only a limited period of time and required repeated injections. PMID- 21647566 TI - Influence of timing of initial surgery for infantile esotropia on the severity of dissociated vertical deviation. AB - PURPOSE: The purpose of this study was to clarify whether the timing of initial surgery for infantile esotropia contributes to better sensory outcomes and to the severity of dissociated vertical deviation (DVD). METHODS: This retrospective study examined 55 children, aged 8 years and younger, who underwent esotropia surgery. Based on age at the time of the surgery, patients were divided into the very early surgery group (birth to 8 months, N = 14), early surgery group (9-24 months, N = 23) and late surgery group (25 months and older, N = 18). Sensory and motor outcomes of the three groups were statistically evaluated. RESULTS: No significant differences in the incidences of DVD were noted among the three groups at the final visit. However, all DVDs of the very early surgery group were latent, whereas 38.9% of the DVDs for both near and distance of the late surgery group were manifest. Significant differences were found among the three groups for the proportion of patients with manifest DVD, latent DVD and without DVD, and for the incidence of additional surgery for manifest DVD (p < 0.05, G-test). CONCLUSION: Early surgery for infantile esotropia decreases the severity of DVD and lowers the need for an additional operation for DVD. PMID- 21647567 TI - Long-term therapeutic efficacy of the subthreshold micropulse diode laser photocoagulation for diabetic macular edema. AB - PURPOSE: To investigate the long-term efficacy of subthreshold micropulse diode laser photocoagulation (SMDLP) on diabetic macular edema (DME). METHODS: The effects of SMDLP (810 nm) were studied on 56 eyes of 44 patients with DME. Optical coherence tomography-determined foveal thickness and best-corrected visual acuity (BCVA) were evaluated at 1, 3, 6, and 12 months after the SMDLP. RESULTS: The mean foveal thickness was 504 MUm before the SMDLP, and it was significantly reduced to 439 MUm at 1 month (P = 0.001), 409 MUm at 3 months (P < 0.0001), 358 MUm at 6 months (P < 0.0001), and 320 MUm at 12 months (P < 0.0001). The mean BCVA at baseline was 0.47 logarithm of the minimal angle of resolution (logMAR) units, and it was not changed significantly at all of the postoperative examinations. Among the 56 eyes, 10 (17.8%) had an improvement of BCVA of >0.2 logMAR units, 36 (64.2%) remained the same, and 10 eyes (17.8%) had a reduction of >0.2 logMAR units at 12 months postoperatively. CONCLUSION: Our results indicate that SMDLP has a beneficial effect on DME. However, prospective studies are needed to evaluate this technique in more detail. PMID- 21647568 TI - Tissue-type plasminogen activator-assisted drainage of suprachoroidal hemorrhage showing a kissing configuration. PMID- 21647569 TI - Eye patching as a treatment for amblyopia in children aged 10-16 years. AB - PURPOSE: To investigate the effects of full-time patching regimen on the treatment of amblyopia in children aged 10-16 years. METHODS: Forty-seven patients with a mean age of 12.09 +/- 1.65 years were included in this study. All of the patients received eye patching for the entire day, 6 days a week, during the first 3 months. The patients who achieved visual acuity of 0.00 logMAR at the third month were provided with additional patching treatment (4-6 h/day). On the other hand, the patients who showed no change in their visual acuity or an increase of less than 0.00 logMAR at the third month had 3 more months of eye patching for the entire day, 6 days a week. RESULTS: Prior to treatment, the best mean visual acuity of the amblyopic eyes was 0.48 +/- 0.25 (range 1.00-0.15) logMAR. After follow-up, the visual acuity of the amblyopic eyes was 0.20 +/- 0.22 (range 1.00-0.00) logMAR. Thus, visual acuity in the amblyopic eyes improved by 0.2 log unit or more in 38 of 47 patients (81%). CONCLUSIONS: The present results show that patching in older children with amblyopia improves visual acuity with no serious complications. The use of patching in children to improve amblyopia seems promising. PMID- 21647570 TI - Classification of acute encephalopathy in respiratory syncytial virus infection. AB - Infection with respiratory syncytial virus (RSV) is known to be associated with central nervous system symptoms such as convulsions. We investigated cytokines, nitrogen oxide (NO)( x ), and the viral genome in cerebrospinal fluid (CSF) obtained from children with RSV infection-related convulsions or central nervous symptoms and compared the data with type of encephalopathy. Of nine patients enrolled (six boys and three girls; aged 10 days-3 years), one metabolic error, five excitotoxicity, one cytokine storm, and two hypoxia cases were found. The patients presented with unilateral convulsions, generalized convulsions, and convulsions following cardiopulmonary arrest, apnea, and nuchal rigidity. In all patients, a rapid check for RSV of nasal fluid was positive. The RSV genome (subgroup A) was detected in the CSF of five of the nine patients; two patients with hypoxic encephalopathy were negative for the RSV genome. The CSF interleukin (IL)-6 levels were high only in patients with the excitotoxicity and cytokine storm type of encephalopathy. NO( x ) levels were high in all the subject cases. In the excitotoxicity type, NO( x ) levels were significantly higher than those in the control and other groups. NO( x ) level may become an important parameter for the diagnosis and classification of acute encephalopathy in RSV. Strategies to treat each type of encephalopathy, targeting cytokines and free radicals, should be established. PMID- 21647571 TI - MEM spectral analysis for predicting influenza epidemics in Japan. AB - OBJECTIVES: The prediction of influenza epidemics has long been the focus of attention in epidemiology and mathematical biology. In this study, we tested whether time series analysis was useful for predicting the incidence of influenza in Japan. METHODS: The method of time series analysis we used consists of spectral analysis based on the maximum entropy method (MEM) in the frequency domain and the nonlinear least squares method in the time domain. Using this time series analysis, we analyzed the incidence data of influenza in Japan from January 1948 to December 1998; these data are unique in that they covered the periods of pandemics in Japan in 1957, 1968, and 1977. RESULTS: On the basis of the MEM spectral analysis, we identified the periodic modes explaining the underlying variations of the incidence data. The optimum least squares fitting (LSF) curve calculated with the periodic modes reproduced the underlying variation of the incidence data. An extension of the LSF curve could be used to predict the incidence of influenza quantitatively. CONCLUSIONS: Our study suggested that MEM spectral analysis would allow us to model temporal variations of influenza epidemics with multiple periodic modes much more effectively than by using the method of conventional time series analysis, which has been used previously to investigate the behavior of temporal variations in influenza data. PMID- 21647572 TI - Stopping, goal-conflict, trait anxiety and frontal rhythmic power in the stop signal task. AB - The medial right frontal cortex is implicated in fast stopping of an initiated motor action in the stop-signal task (SST). To assess whether this region is also involved in the slower behavioural inhibition induced by goal conflict, we tested for effects of goal conflict (when stop and go tendencies are balanced) on low frequency rhythms in the SST. Stop trials were divided, according to the delays at which the stop signal occurred, into short-, intermediate-, and long-delay trials. Consistent with goal-conflict processing, intermediate-delay trials were associated with greater 7-8 Hz EEG power than short- or long-delay trials at medial right frontal sites (Fz, F4, and F8). At F8, 7-8 Hz power was linked to high trait anxiety and neuroticism. A separate 4-7 Hz power increase was also seen in stop, relative to go, trials, but this was independent of delay, was maximal at the central midline site Cz, and predicted faster stopping. Together with previous data on the SST, these results suggest that the right frontal region could be involved in multiple inhibition mechanisms. We propose a hierarchical model of the control of stopping that integrates the literature on the neural control of fast motor stopping with that on slower, motive-directed behavioural inhibition. PMID- 21647573 TI - Osteoporosis: a paradox in ankylosing spondylitis. AB - Ankylosing spondylitis is a chronic and severe inflammatory disease of the axial skeleton and the joints. Inflammation is associated with trabecular bone loss leading to osteoporosis but also with corcal new bone formation leading to progressive ankylosis of the spine and sacroiliac joints. This results in an apparent paradox of bone formation and loss taking place at sites closesly located to each other. Osteoporosis can be explained by the impact of inflammation of the bone remodeling cycle. In contrast, new bone formation has been linked to aberrant acvaon of bone morphogenec protein and Wnt signaling. In this commentary, we review recent data on this bone paradox and highlight recent advances including the effect of current drug therapies and the idenfication of new therapeutic targets. PMID- 21647574 TI - Prediction of stroke risk in atrial fibrillation, prevention of stroke in atrial fibrillation, and the impact of long-term monitoring for detecting atrial fibrillation. AB - Atrial fibrillation (AF) is a large public health problem that affects about 1% of the population in the United States. It confers an increased risk for stroke and thromboembolism, but the stroke risk is not equal in all patients. Further refinement in stratifying stroke risk in patients with AF will help in properly directing therapy for AF patients while minimizing adverse events. Warfarin is the first-line treatment for stroke reduction in patients with AF, but many new drugs are on the horizon that will significantly change practice. New and improved cardiac monitoring techniques and devices will help with detection of AF in those at risk for stroke and will assist in assessing which patients will most benefit from anticoagulation. PMID- 21647575 TI - Role of coronary artery calcium score and coronary CT angiography in the diagnosis and risk stratification of individuals with suspected coronary artery disease. AB - Coronary heart disease is the leading cause of death worldwide, and has traditionally been assessed through a patient's cardiovascular risk profile that is comprised of a combination of genetic, social, physiologic, and environmental factors. A growing discordance is being recognized in the ability of current risk estimation tools to predict outcomes versus that of actual measured outcomes. Exciting new improvements in technology have made noninvasive imaging modalities of the heart--in particular, coronary artery calcium score (CACS) and coronary computed tomography (CT) angiography--an increasingly important component in the diagnosis of ischemic heart disease. The CACS has been found to be a marker of vascular injury that correlates closely with overall atherosclerotic burden, whereas coronary CT angiography permits detection of noncalcified plaque coronary artery stenosis severity. A growing body of literature has developed detailing the valuable prognostic utility of these tests in the management of patients and how they may 1 day be used to complement current risk prediction models. PMID- 21647576 TI - Perioperative care of children with tetralogy of fallot. AB - OPINION STATEMENT: Complete surgical repair of tetralogy of Fallot (TOF) around 3 months of age is the commonly undertaken management strategy in many centers and has excellent outcomes. Intervention at an earlier age, including the newborn period, may be required for children with symptoms. Early extubation from mechanical ventilation where possible may help improve outcomes for children undergoing complete repair of TOF. PMID- 21647577 TI - Psychoactive drugs and false memory: comparison of dextroamphetamine and delta-9 tetrahydrocannabinol on false recognition. AB - RATIONALE: Several psychoactive drugs are known to influence episodic memory. However, these drugs' effects on false memory, or the tendency to incorrectly remember nonstudied information, remain poorly understood. OBJECTIVES: Here, we examined the effects of two commonly used psychoactive drugs, one with memory enhancing properties (dextroamphetamine; AMP), and another with memory-impairing properties (Delta(9)-tetrahydrocannabinol; THC), on false memory using the Deese/Roediger-McDermott (DRM) illusion. METHODS: Two parallel studies were conducted in which healthy volunteers received either AMP (0, 10, and 20 mg) or THC (0, 7.5, and 15 mg) in within-subjects, randomized, double-blind designs. Participants studied DRM word lists under the influence of the drugs, and their recognition memory for the studied words was tested 2 days later, under sober conditions. RESULTS: As expected, AMP increased memory of studied words relative to placebo, and THC reduced memory of studied words. Although neither drug significantly affected false memory relative to placebo, AMP increased false memory relative to THC. Across participants, both drugs' effects on true memory were positively correlated with their effects on false memory. CONCLUSIONS: Our results indicate that AMP and THC have opposing effects on true memory, and these effects appear to correspond to similar, albeit more subtle, effects on false memory. These findings are consistent with previous research using the DRM illusion and provide further evidence that psychoactive drugs can affect the encoding processes that ultimately result in the creation of false memories. PMID- 21647578 TI - Translational PK-PD modelling of molecular target modulation for the AMPA receptor positive allosteric modulator Org 26576. AB - INTRODUCTION: The alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor potentiator Org 26576 represents an interesting pharmacological tool to evaluate the utility of glutamatergic enhancement towards the treatment of psychiatric disorders. In this study, a rat-human translational pharmacokinetic pharmacodynamic (PK-PD) model of AMPA receptor modulation was used to predict human target engagement and inform dose selection in efficacy clinical trials. METHODS: Modelling and simulation was applied to rat plasma and cerebrospinal fluid (CSF) pharmacokinetic and pharmacodynamic measurements to identify a target concentration (EC(80)) for AMPA receptor modulation. Human plasma pharmacokinetics was determined from 33 healthy volunteers and eight major depressive disorder patients. From four out of these eight patients, CSF PK was also determined. Simulations of human CSF levels were performed for several doses of Org 26576. RESULTS: Org 26576 (0.1-10 mg/kg, i.v.) potentiated rat hippocampal AMPA receptor responses in an exposure-dependant manner. The rat plasma and CSF PK data were fitted by one-compartment model each. The rat CSF PK-PD model yielded an EC(80) value of 593 ng/ml (90% confidence interval 406.8, 1,264.1). The human plasma and CSF PK data were simultaneously well described by a two compartment model. Simulations showed that in humans at 100 mg QD, CSF levels of Org 26576 would exceed the EC(80) target concentration for about 2 h and that 400 mg BID would engage AMPA receptors for 24 h. CONCLUSION: The modelling approach provided useful insight on the likely human dose-molecular target engagement relationship for Org 26576. Based on the current analysis, 100 and 400 mg BID would be suitable to provide 'phasic' and 'continuous' AMPA receptor engagement, respectively. PMID- 21647579 TI - Antidepressant-like properties of sarizotan in experimental Parkinsonism. AB - RATIONALE: Depression and anxiety are common symptoms in Parkinson's disease for which there are no optimal treatments. Sarizotan, an agonist at serotonin receptors and partial agonist at dopamine D2-like receptors, has shown antidyskinetic effects in Parkinson's disease. Based on its pharmacological profile, we hypothesized that sarizotan could also have antidepressant-like properties. OBJECTIVES: Examine effects of sarizotan on behavioral and histological measures known to be regulated by established antidepressants in normal and unilaterally 6-hydroxydopamine-lesioned rats. RESULTS: Sarizotan was found to significantly reduce immobility in the modified forced swim test, a measure of antidepressant-like activity, but had no effects on thigmotaxis or corner time, measures of anxiety-like behavior, in the unilaterally 6 hydroxydopamine-lesioned rats. At the same dose, sarizotan counteracted L: DOPA/benserazide-induced supersentitized rotational behavior and dyskinesias without significantly affecting L: -DOPA/benserazide-induced locomotion. At the histological level, sarizotan alone or in combination with L: -DOPA/benserazide stimulated cell proliferation, measured by bromodeoxyuridine incorporation or Ki 67 staining, both in the subgranular zone of the dentate gyrus and in the subventricular zone of the striatum in the 6-hydroxydopamine-lesioned hemisphere. Likewise, combined sarizotan and L: -DOPA/benserazide treatment stimulated doublecortin levels in the subgranular zone of the dentate gyrus. CONCLUSIONS: These significant effects of sarizotan in the modified forced swim test and on cell proliferation are reminiscent of those found after various antidepressant therapies. These data suggest that sarizotan may have some antidepressant-like and restorative properties in Parkinsonism. PMID- 21647580 TI - Anti-ribosomal P antibodies and lupus nephritis. AB - Increasing attention has been paid to the relationship of autoantibodies to ribosomal P proteins (anti-P) with lupus nephritis. Several mechanisms of involvement of anti-P in lupus nephritis have been proposed, including cross reactivity with anti-dsDNA and anti-endothelial cell antibodies (AECA). In addition, it is also suggested that anti-P might play a role in the development of lupus nephritis through induction of T helper 1 responses. Of note, recent studies have disclosed that the presence of isolated anti-P antibodies may discriminate patients with pure class V lupus nephritis, whereas the simultaneous presence of anti-dsDNA antibodies suggests class V disease with concomitant proliferative lesions. These observations lead to the hypothesis that anti-P and anti-dsDNA might result in membraneous changes and proliferative changes, respectively, although further investigation is required for confirmation. PMID- 21647581 TI - Lenalidomide is active for extramedullary disease in refractory multiple myeloma. PMID- 21647582 TI - Mimicry of hydroxyurea-induced leg ulcer by distal vena saphena parva insufficiency. PMID- 21647583 TI - Low absolute lymphocyte count and addition of rituximab confer high risk for interstitial pneumonia in patients with diffuse large B-cell lymphoma. AB - Several small-scale studies have reported pulmonary toxicity among patients with diffuse large B-cell lymphoma (DLBCL) receiving rituximab-containing chemotherapy, though whether the use of rituximab predisposes to interstitial pneumonia (IP) remains unclear. This retrospective study was intended to identify the characteristics and risk factors of IP in patients with DLBCL. Between 2000 and 2009, 529 consecutive patients with DLBCL receiving first-line tri-weekly COP or CHOP-based chemotherapy with or without rituximab were enrolled as subjects. IP was defined as diffuse pulmonary interstitial infiltrates found on computed tomography scans in conjunction with respiratory symptoms. IP was observed in 26 patients (4.9%), six of whom were confirmed with Pneumocystis jirovecii pneumonia. The median number of chemotherapy courses before IP was four cycles. Using multivariate analysis, absolute lymphocyte count less than 1*10(9)/l at diagnosis [odds ratio (OR) 2.75, p=0.014] and the addition of rituximab to chemotherapy (OR 4.56, p=0.003) were identified as independent risk factors for IP. In conclusion, the incidence of IP is increased in patients with DLBCL receiving rituximab-containing chemotherapy. Specific subgroups with lymphopenia at diagnosis may justify close scrutiny to detect pulmonary complications. PMID- 21647584 TI - Lung injury in a leukemia patient during mobilization of peripheral blood stem cells using granulocyte colony-stimulating factor alone. PMID- 21647585 TI - Promoting the effect of chemical constituents from the flowers of Poacynum hendersonii on adipogenesis in 3T3-L1 cells. AB - A novel sugar ester poacynose (1) was isolated from the flowers of Poacynum hendersonii together with 31 known compounds. The structure of 1 was established mainly on the basis of 1D and 2D NMR spectral data. Among the isolates, several constituents, such as kaempferol 3-O-sophoroside (5), picein (16), and 4 hydroxybenzoic acid 4-O-beta-D-glucopyranoside (17) moderately promoted adipogenesis of 3T3-L1 cells. In addition, simultaneous quantitative analysis of eight flavonoid constituents from the flower and leaf parts of P. hendersonii was developed. PMID- 21647586 TI - Identification of potential microRNAs and their targets in Brassica rapa L. AB - MicroRNAs (miRNAs) are recently discovered, noncoding, small regulatory RNA molecules that negatively regulate gene expression. Although many miRNAs are identified and validated in many plant species, they remain largely unknown in Brassica rapa (AA 2n =, 20). B. rapa is an important Brassica crop with wide genetic and morphological diversity resulting in several subspecies that are largely grown for vegetables, oilseeds, and fodder crop production. In this study, we identified 186 miRNAs belonging to 55 families in B. rapa by using comparative genomics. The lengths of identified mature and pre-miRNAs ranged from 18 to 22 and 66 to 305 nucleotides, respectively. Comparison of 4 nucleotides revealed that uracil is the predominant base in the first position of B. rapa miRNA, suggesting that it plays an important role in miRNA-mediated gene regulation. Overall, adenine and guanine were predominant in mature miRNAs, while adenine and uracil were predominant in pre-miRNA sequences. One DNA sequence producing both sense and antisense mature miRNAs belonging to the BrMiR 399 family, which differs by 1 nucleotide at the, 20(th) position, was identified. In silico analyses, using previously established methods, predicted 66 miRNA target mRNAs for 33 miRNA families. The majority of the target genes were transcription factors that regulate plant growth and development, followed by a few target genes that are involved in fatty acid metabolism, glycolysis, biotic and abiotic stresses, and other cellular processes. Northern blot and qRT-PCR analyses of RNA samples prepared from different B. rapa tissues for 17 miRNA families revealed that miRNAs are differentially expressed both quantitatively and qualitatively in different tissues of B. rapa. PMID- 21647587 TI - Assessment of pituitary adenoma volumetric change using longitudinal MR image registration. AB - INTRODUCTION: Change detection is a crucial factor in monitoring of slowly evolving pathologies. The objective of the study was to test a semi-automatic method applied on longitudinal MRI monitoring of volume change in pituitary macroadenomas. METHODS: The proposed method is based on a visual comparison of geometrically corrected, co-registered, intensity-normalized contrast-enhanced (CE) 3D GRE T1-weighted images. Qualitative volume changes based on this applied method were compared with experts' readings of conventional pre- and post-CE 2D T1-weighted images. Magnetic resonance (MR) imaging was performed two to four times in 13 patients with a total combination of 29 time points. RESULTS: Compared to conventional 2D MR readings, a diagnosis of tumor growth (yes/no) was changed in 5 of 13 patients (38%) at 9 of the 29 combinations of time points (31%) using the 3D-based semi-automatic method. With manual tumor tracings as reference, McNemar's test showed a significant difference between the two methods. CONCLUSION: Visual comparison of geometrically corrected, intensity normalized, and affine-aligned longitudinal 3D images may enable more accurate assessment of qualitative volumetric change in pituitary adenomas than conventional reading of 2D images. PMID- 21647588 TI - Correlation between maxillofacial radiographic features and systemic severity as sickle cell disease severity predictor. AB - This study was conducted to investigate the relationship among radiographic features observed on panoramic radiographs of sickle cell disease patients and analyze their relationship with history of systemic severity of the disease. Panoramic radiographs of 71 subjects with sickle cell disease were evaluated for the presence of the following radiographic bony alterations: radiopaque areas, increased spacing of bony trabeculae, horizontal arrangement of bony trabeculae and corticalization of mandibular canal. History of clinical systemic severity was assessed through direct questioning about the frequency of vaso-occlusive crisis, history of stroke, clinical jaundice, femur head necrosis, and leg ulceration. Chi-square or Fisher's exact test were applied in order to analyze possible associations between radiographic features and history of complications, with p < 0.05 significance level. Increased spacing of bony trabeculae was statistically associated with absence of corticalization of mandibular canal (p < 0.01) and horizontal arrangement of bony trabeculae (p = 0.04). Statistically significant associations were demonstrated between history of clinical jaundice and presence of increased spacing of bony trabeculae (p = 0.02) and between history of stroke and presence of horizontal arrangement of bony trabeculae (p = 0.04). Based on the results of the current study, maxillofacial radiographic features may be associated with clinical parameters of systemic complications in sickle cell disease patients. The relationship between radiographic features and history of complications associated with clinical severity of sickle cell disease has not been demonstrated in the literature. Acknowledgment of such possible association may help establish prognosis and influence clinical treatment of systemic and oral complications. PMID- 21647589 TI - Comparison of the push-out strength of two fiber post systems dependent on different types of resin cements. AB - The purpose of this study was to compare the push-out strength of glass fiber posts dependent on the resin cement. One hundred human teeth were divided into five groups (n = 20). Two glass fiber post systems (DT Light SL (DTSL) and RelyX Fiber Post (RF)) were used. DTSL posts were cemented with one "etch & rinse" system (ER) or one of three self-adhesive resin cements (SA). The RF posts were cemented with RelyX Unicem. Afterwards, half of the specimens were thermocycled (TC; 5 degrees C/55 degrees C, 5,000 cycles). All specimens were cut into disks (thickness, 2 mm). The push-out test was performed (crosshead speed, 1 mm/min), fracture types were determined (*25 and *40 magnification), and statistical analysis was performed (one-way analysis of variance (ANOVA), Scheffe test, p < 0.05). One-way ANOVA showed a significant influence of the resin cement on the push-out strength of the glass fiber posts before thermocycling (p < 0.001). After TC, no significant differences were detected. Microscopic evaluation showed mainly adhesive failures between post and cement for ER or mixed fractures for SA. The bond strength of adhesively cemented glass fiber posts is not dependent on the type of resin cement after TC. The use of SA can lead to bond strength values comparable to ER. Self-adhesive resin cements could be used just as well as resin cements with "etch & rinse" adhesive systems for the cementation of glass fiber posts. PMID- 21647590 TI - Human periodontal ligament fibroblasts stimulated by nanocrystalline hydroxyapatite paste or enamel matrix derivative. An in vitro assessment of PDL attachment, migration, and proliferation. AB - We determined the effects of soluble or coated nanocrystalline hydroxyapatite paste (nano-HA) and enamel matrix derivative (EMD) on proliferation, adhesion, and migration of periodontal ligament fibroblasts (PDLs). Cultured PDLs were stimulated with nano-HA paste or EMD in a soluble form or were coated to the surface of cell culture dishes. Proliferation of PDLs on coated nano-HA and EMD was quantified by various methods including bromodeoxyuridine (BrdU) incorporation and Western blot. Cell migration was investigated in a modified Boyden chamber. The surface integrin profile of PDLs was determined using an integrin-specific ELISA, and integrin-specific signaling was measured by immunoblotting of phosphorylated focal adhesion kinase (FAK). Coated nano-HA stimulated PDL proliferation to a larger extent as compared with coated EMD. PDL migration towards a nano-HA or EMD gradient was more efficiently mediated by soluble EMD as compared with nano-HA but vice versa, adhesion of PDLs to compound coated dishes was more effectively mediated by nano-HA as compared with EMD. Mechanistically, majorly integrin alpha5beta1-mediated adhesion of PDL and both coated compounds mediated a significant increase in FAK activation though to a different extent. Current findings offer two different modes of action for EMD and nano-HA paste. EMD efficiently acts as a chemoattractant in its soluble form, while nano-HA paste effectively serves as a synthetic extracellular matrix component in its coated form. Our findings suggest that EMD and nano-HA paste display different molecular characteristics and apply alternative routes to mediate their beneficial effects on periodontal tissues. PMID- 21647591 TI - Digital evaluation of the reproducibility of implant scanbody fit--an in vitro study. AB - Dental restorations are increasingly manufactured by CAD/CAM systems. Currently, there are two alternatives for digitizing dental implants: direct intra-oral data capturing or indirect from a master cast, both with transfer caps (scanbodies). The aim of this study was the evaluation of the fit of the scanbodies and their ability of reposition. At the site of the first molars and canines, implants were placed bilaterally in a polymer lower arch model (original model), and an impression was taken for fabricating a stone cast (stone model). Ten white-light scans were obtained from the original and the stone model with the scanbodies in place. The scanbodies were retrieved after each scan and re-attached to the same implant or lab analogue. The first scan of the series served as control in both groups. The subsequent nine scans and control were superimposed using inspection software to identify the discrepancies of the four scanbodies in both experimental groups. The systematic error of digitizing the models was 13 MUm for the polymer and 5 MUm for the stone model. The mean discrepancy of the scanbodies was 39 MUm (+/-58 MUm) on the original implants versus 11 MUm (+/-17 MUm) on the lab analogues. The difference in scanbody discrepancy between original implants and lab analogues was statistically significant (p < 0.05, Mann-Whitney U test). Scanbody discrepancy was higher on original implants than on lab analogues. Fit and reproducibility of the scanbodies on original implants should be improved to achieve higher accuracy of implant-supported CAD/CAM fabricated restorations. PMID- 21647592 TI - CFTR induces extracellular acid sensing in Xenopus oocytes which activates endogenous Ca2+-activated Cl- conductance. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) produces a cyclic adenosine monophosphate (cAMP)-dependent Cl- conductance of distinct properties that is essential for electrolyte secretion in human epithelial tissues. However, the functional consequences of CFTR expression are multifaceted, encompassing much more than simply supplying a cellular cAMP-regulated Cl- conductance. When we expressed CFTR in Xenopus oocytes, we found that extracellular acidic pH activates a Ca2+-dependent outwardly rectifying Cl- conductance that does not reflect CFTR activity. The proton-activated Cl- conductance showed biophysical and pharmacological features of a Ca2+-dependent Cl- conductance, most likely mediated by Xenopus TMEM16A. In contrast to the effects of extracellular acidification, intracellular acidification did not activate an endogenous Cl- conductance. Proton/CFTR-mediated activation of human TMEM16A was also detected in HEK293 cells. The gating mutant G551D-CFTR conferred proton sensitivity, while deltaF508-CFTR enabled proton activation of TMEM16A only in Xenopus oocytes, which, unlike HEK293 cells, allow deltaF508-CFTR to be trafficked to the cell membrane. Activation of TMEM16A by lysophosphatidic acid was enhanced in the presence of CFTR but was additive with activation by extracellular protons. Because expression of CFTR-E1474X did not confer proton sensitivity, we propose that CFTR translocates a proton receptor to the plasma membrane via its PDZ binding domain. PMID- 21647593 TI - Epithelial-to-mesenchymal transition in podocytes mediated by activation of NADPH oxidase in hyperhomocysteinemia. AB - The present study tested the hypothesis that hyperhomocysteinemia (hHcys) induces podocytes to undergo epithelial-to-mesenchymal transition (EMT) through the activation of NADPH oxidase (Nox). It was found that increased homocysteine (Hcys) level suppressed the expression of slit diaphragm-associated proteins, P cadherin and zonula occludens-1 (ZO-1), in conditionally immortalized mouse podocytes, indicating the loss of their epithelial features. Meanwhile, Hcys remarkably increased the abundance of mesenchymal markers, such as fibroblast specific protein-1 (FSP-1) and alpha-smooth muscle actin (alpha-SMA). These phenotype changes in podocytes induced by Hcys were accompanied by enhanced superoxide (O-2) production, which was substantially suppressed by inhibition of Nox activity. Functionally, Hcys significantly enhanced the permeability of the podocyte monolayer coupled with increased EMT, and this EMT-related increase in cell permeability could be restored by Nox inhibitors. In mice lacking gp91( phox ) (gp91(-/-)), an essential Nox subunit gene, hHcys-enhanced podocyte EMT and consequent glomerular injury were examined. In wild-type (gp91(+/+)) mice, hHcys induced by a folate-free diet markedly enhanced expression of mesenchymal markers (FSP-1 and alpha-SMA) but decreased expression of epithelial markers of podocytes in glomeruli, which were not observed in gp91(-/-) mouse glomeruli. Podocyte injury, glomerular sclerotic pathology, and marked albuminuria observed in gp91(+/+) mice with hHcys were all significantly attenuated in gp91(-/-) mice. These results suggest that hHcys induces EMT of podocytes through activation of Nox, which represents a novel mechanism of hHcys-associated podocyte injury. PMID- 21647594 TI - Authorship and publication practices in the social sciences: historical reflections on current practices. AB - An historical review of authorship definitions and publication practices that are embedded in directions to authors and in the codes of ethics in the fields of psychology, sociology, and education illuminates reasonable agreement and consistency across the fields with regard to (a) originality of the work submitted, (b) data sharing, (c) human participants' protection, and (d) conflict of interest disclosure. However, the role of the professional association in addressing violations of research or publication practices varies among these fields. Psychology and sociology provide active oversight with sanction authority. In education, the association assumes a more limited role: to develop and communicate standards to evoke voluntary compliance. With respect to authorship credit, each association's standards focus on criteria for inclusion as an author, other than on the author's ability to defend and willingness to take responsibility for the entire work. Discussions across a broad range of research disciplines beyond the social sciences would likely be beneficial. Whether improved standards will reduce either misattribution or perceptions of inappropriate attribution of credit within social science disciplines will likely depend on how well authorship issues are addressed in responsible conduct of research education (RCR), in research practice, and in each association's ongoing efforts to influence normative practice by specifying and clarifying best practices. PMID- 21647595 TI - Assessing the preparedness of research integrity officers (RIOs) to appropriately handle possible research misconduct cases. AB - Institutions receiving federal funding for research from the U.S.Public Health Service need to have policies and procedures to both prevent research misconduct and to adjudicate it when it occurs. The person who is designated to handle research misconduct is typically referred to as the research integrity officer (RIO). In this interview study we report on 79 RIOs who describe how they would handle allegations of research misconduct. Their responses were compared to two expert RIOs. The responses to the allegations in the scenarios demonstrated that RIOs are not uniformly well prepared to handle activities associated with reported allegations of research misconduct. We recommend greater preparation through directed training, use of check lists of possible behaviors necessary to consider when situations arise, being involved in a network of RIOs so one can discuss options, and the possible need to certify RIOs. PMID- 21647597 TI - Uterine packing during cesarean section in the management of intractable hemorrhage in central placenta previa. AB - OBJECTIVE: To determine the safety and effectiveness of uterine packing in the management of intractable hemorrhage during cesarean section for central placenta previa. METHODS: This retrospective study was conducted on 70 pregnant women with central placenta previa from May 2005 to March 2010. Patients with uterine packing in the control of massive hemorrhage during cesarean section were identified. The indications, uterine packing procedures, estimated blood loss, postoperative complications, and packing material used were reviewed. RESULTS: A total of 70 patients were identified among 1,121 women with placenta previa during the study period. Sixty-five cases were successful in the control of intraoperative bleeding using uterine packing. Two patients with severe placenta accreta had hemorrhage during cesarean section, and packing with gauze in the uterine cavity was not able to control the bleeding, thereby resulting in cesarean hysterectomy. One case demonstrated failure in packing because of disseminated intravascular coagulation occurring before hospital admission. The remaining two patients had massive vaginal bleeding after uterine packing in cesarean section and underwent laparotomy or hysterectomy 4 h postoperative. CONCLUSION: Uterine packing is a safe and effective technique in the control of intractable hemorrhage in cesarean section. It is a reasonable alternative to further surgical intervention in patients with intractable obstetric hemorrhage, especially in developing countries. PMID- 21647596 TI - Benign external hydrocephalus: a review, with emphasis on management. AB - Benign external hydrocephalus in infants, characterized by macrocephaly and typical neuroimaging findings, is considered as a self-limiting condition and is therefore rarely treated. This review concerns all aspects of this condition: etiology, neuroimaging, symptoms and clinical findings, treatment, and outcome, with emphasis on management. The review is based on a systematic search in the Pubmed and Web of Science databases. The search covered various forms of hydrocephalus, extracerebral fluid, and macrocephaly. Studies reporting small children with idiopathic external hydrocephalus were included, mostly focusing on the studies reporting a long-term outcome. A total of 147 studies are included, the majority however with a limited methodological quality. Several theories regarding pathophysiology and various symptoms, signs, and clinical findings underscore the heterogeneity of the condition. Neuroimaging is important in the differentiation between external hydrocephalus and similar conditions. A transient delay of psychomotor development is commonly seen during childhood. A long-term outcome is scarcely reported, and the results are varying. Although most children with external hydrocephalus seem to do well both initially and in the long term, a substantial number of patients show temporary or permanent psychomotor delay. To verify that this truly is a benign condition, we suggest that future research on external hydrocephalus should focus on the long-term effects of surgical treatment as opposed to conservative management. PMID- 21647598 TI - Increased expression of GAPDH protein is not indicative of nitrosative stress or apoptosis in liver of starved rainbow trout (Oncorhynchus mykiss). AB - Short-term starvation has been linked to in vivo protein degradation in liver of rainbow trout (Oncorhynchus mykiss). However, it is unclear whether this proposed increase in protein degradation is followed by programmed cell death (apoptosis) in liver of starved trout. A preliminary study in our laboratory revealed an isoform of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein that increased 4.5-fold in liver of starved trout. GAPDH is a glycolytic enzyme involved in other cellular functions, including apoptosis. Increased intracellular nitric oxide (NO) promotes nuclear translocation of GAPDH that is associated with increased apoptosis in mammals. If GAPDH protein is associated with apoptosis in rainbow trout, it could potentially be used as a biomarker of cellular stress in liver of teleost fish species. The purpose of this study was to determine whether increased GAPDH protein expression in liver of starved rainbow trout is associated with NO-induced apoptosis. Targeted proteomic analysis using multiple reaction monitoring (MRM) was used to determine the level of GAPDH in nuclear and cytoplasmic fractions and inducible nitric oxide synthase (iNOS) in cell lysates. Dot blot and DNA fragmentation analyses were conducted to evaluate protein S-nitrosylation and apoptosis, respectively. Results showed that cytoplasmic GAPDH was 3.4-fold higher in liver of starved versus fed rainbow trout but could not be detected in nuclear fractions. Starvation significantly reduced hepato-somatic index but had no effect on iNOS protein expression, protein S-nitrosylation, or apoptosis. Our results indicate that starvation promoted significant reduction in liver mass that was not associated with increased apoptosis or NO-induced stress and that greater GAPDH concentration in liver of starved rainbow trout was located primarily in the cytoplasm. PMID- 21647599 TI - Migration and chemokine receptor pattern of colitis-preventing DX5+NKT cells. AB - PURPOSE: DX5(+)NKT cells are a subpopulation of NKT cells expressing both T cell receptor and NK cell markers that show an immune-regulating function. Transferred DX5(+)NKT cells from immune competent Balb/c mice can prevent or reduce induced colitis in severe combined immunodeficient (SCID) mice. Here, we investigated the in vivo migration of DX5(+)NKT cells and their corresponding chemokine receptor patterns. METHODS: DX5(+)NKT cells were isolated from spleens of Balb/c mice and transferred into Balb/c SCID mice. After 2 and 8 days, in vivo migration was examined using in vivo microscopy. In addition, the chemokine receptor pattern was analyzed with fluorescence-activated cell sorting (FACS) and the migration assay was performed. RESULTS: Our results show that labeled DX5(+)NKT cells were primarily detectable in mesenteric lymph nodes and spleen after transfer. After 8 days, DX5(+)NKT cells were observed in the colonic tissues, especially the appendix. FACS analysis of chemokine receptors in DX5(+)NKT cells revealed expression of CCR3, CCR6, CCR9, CXCR3, CXCR4, and CXCR6, but no CCR5, CXCR5, or the lymphoid homing receptor CCR7. Stimulation upregulated especially CCR7 expression, and chemokine receptor patterns were different between splenic and liver DX5(+)NKT cells. CONCLUSIONS: These data indicate that colitis-preventing DX5(+)NKT cells need to traffic through lymphoid organs to execute their immunological function at the site of inflammation. Furthermore, DX5(+)NKT cells express a specific chemokine receptor pattern with an upregulation of the lymphoid homing receptor CCR7 after activation. PMID- 21647600 TI - Stress generation and adolescent depression: contribution of interpersonal stress responses. AB - This research examined the proposal that ineffective responses to common interpersonal problems disrupt youths' relationships, which, in turn, contributes to depression during adolescence. Youth (86 girls, 81 boys; M age = 12.41, SD = 1.19) and their primary female caregivers participated in a three-wave longitudinal study. Youth completed a measure assessing interpersonal stress responses; youth and caregivers completed semi-structured interviews assessing youths' life stress and psychopathology. Consistent with the hypothesized model, ineffective stress responses (low levels of effortful engagement, high levels of involuntary engagement and disengagement) predicted the generation of subsequent interpersonal stress, which partially accounted for the association between stress responses and depression over time. Moreover, results revealed that self generated interpersonal, but not noninterpersonal stress, predicted depression, and that this explanatory model was specific to the prediction of depression but not anxiety. This research builds on interpersonal stress generation models of depression, and highlights the importance of implementing depression-focused intervention programs that promote effective stress responses and adaptive interpersonal relationships during adolescence. PMID- 21647601 TI - Chronic stress in pregnant guinea pigs (Cavia aperea f. porcellus) attenuates long-term stress hormone levels and body weight gain, but not reproductive output. AB - Stress, when extreme or chronic, can have a negative impact on health and survival of mammals. This is especially true for females during reproduction when self-maintenance and investment in offspring simultaneously challenge energy turnover. Therefore, we investigated the effects of repeated stress during early- and mid-gestation on the maternal stress axis, body weight gain and reproductive output. Female guinea pigs (Cavia aperea f. porcellus, n = 14) were either stressed (treatment: exposure to strobe light in an unfamiliar environment on gestational day -7, 0, 7, 14, 21, 28, 35, 42) or left completely undisturbed (control) throughout pregnancy. Females of both groups received the same respective diets, and reproductive parameters were evaluated upon parturition. Additionally, hormonal data were obtained from blood and feces. The stress exposure induced a significant increase in plasma cortisol concentrations during the afternoon. In contrast to this short-term response in plasma cortisol concentrations, we found no significant differences in the levels of cortisol metabolites in feces collected after stress exposure between groups and even significantly decreased levels of fecal cortisol metabolites on non-stress days over time in treatment females. Among treatment females, gain in body weight was attenuated over gestation and body weight was lower compared to control females during lactation, especially in cases of large litter sizes. No differences could be seen in the reproductive parameters. We conclude that repeated stress exposure with strobe light during early- and mid-gestation results in a down-regulation of the hypothalamic-pituitary-adrenal axis and lower weight gain in treatment females, but has no effect on reproductive output. PMID- 21647602 TI - Type 2 diabetes and obesity: genomics and the clinic. AB - Type 2 diabetes (T2D) and obesity represent major challenges for global public health. They are at the forefront of international efforts to identify the genetic variation contributing to complex disease susceptibility, and recent years have seen considerable success in identifying common risk-variants. Given the clinical impact of molecular diagnostics in rarer monogenic forms of these diseases, expectations have been high that genetic discoveries will transform the prospects for risk stratification, development of novel therapeutics and personalised medicine. However, so far, clinical translation has been limited. Difficulties in defining the alleles and transcripts mediating association effects have frustrated efforts to gain early biological insights, whilst the fact that variants identified account for only a modest proportion of observed familiarity has limited their value in guiding treatment of individual patients. Ongoing efforts to track causal variants through fine-mapping and to illuminate the biological mechanisms through which they act, as well as sequence-based discovery of lower-frequency alleles (of potentially larger effect), should provide welcome acceleration in the capacity for clinical translation. This review will summarise recent advances in identifying risk alleles for T2D and obesity, and existing contributions to understanding disease pathology. It will consider the progress made in translating genetic knowledge into clinical utility, the challenges remaining, and the realistic potential for further progress. PMID- 21647604 TI - Characterization of PSI recovery after chilling-induced photoinhibition in cucumber (Cucumis sativus L.) leaves. AB - By simultaneously analyzing the chlorophyll a fluorescence transient and light absorbance at 820 nm as well as chlorophyll fluorescence quenching, we investigated the effects of different photon flux densities (0, 15, 200 MUmol m( 2) s(-1)) with or without 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) on the repair process of cucumber (Cucumis sativus L.) leaves after treatment with low temperature (6 degrees C) combined with moderate photon flux density (200 MUmol m(-2 )s(-1)) for 6 h. Both the maximal photochemical efficiency of Photosystem II (PSII) (F (v)/F (m)) and the content of active P700 (DeltaI/I (o)) significantly decreased after chilling treatment under 200 MUmol m(-2 )s(-1) light. After the leaves were transferred to 25 degrees C, F (v)/F (m) recovered quickly under both 200 and 15 MUmol m(-2 )s(-1) light. DeltaI/I (o) recovered quickly under 15 MUmol m(-2) s(-1) light, but the recovery rate of DeltaI/I (o) was slower than that of F (v)/F (m). The cyclic electron transport was inhibited by chilling-light treatment obviously. The recovery of DeltaI/I (o) was severely suppressed by 200 MUmol m(-2) s(-1) light, whereas a pretreatment with DCMU effectively relieved this suppression. The cyclic electron transport around PSI recovered in a similar way as the active P700 content did, and the recovery of them was both accelerated by pretreatment with DCMU. The results indicate that limiting electron transport from PSII to PSI protected PSI from further photoinhibition, accelerating the recovery of PSI. Under a given photon flux density, faster recovery of PSII compared to PSI was detrimental to the recovery of PSI or even to the whole photosystem. PMID- 21647603 TI - XCI in preimplantation mouse and human embryos: first there is remodelling.... AB - Female eutherians silence one of their X chromosomes to accomplish an equal dose of X-linked gene expression compared with males. The mouse is the most widely used animal model in XCI research and has proven to be of great significance for understanding the complex mechanism of X-linked dosage compensation. Although the basic principles of XCI are similar in mouse and humans, differences exist in the timing of XCI initiation, the genetic elements involved in XCI regulation and the form of XCI in specific tissues. Therefore, the mouse has its limitations as a model to understand early human XCI and analysis of human tissues is required. In this review, we describe these differences with respect to initiation of XCI in human and mouse preimplantation embryos, the extra-embryonic tissues and the in vitro model of the epiblast: the embryonic stem cells. PMID- 21647605 TI - Association mapping of dynamic developmental plant height in common wheat. AB - Drought as a major abiotic stress often occurs from stem elongation to the grain filling stage of wheat in northern China. Plant height (PH) is a suitable trait to model the dissection of drought tolerance. The purposes of the present study were to validate molecular markers for PH developmental behavior and identify elite alleles of molecular markers. After the phenotyping of 154 accessions for PH dynamic development under well-watered (WW) and drought stressed (DS) conditions, and the genotyping of 60 SSR markers from six candidate chromosome regions related to PH found in our previous linkage mapping studies, both parameters PH and drought tolerance coefficient (DTC) calculated by the conditional analysis were used for association mapping. A total of 46 significant association signals (P < 0.01) were identified in 23 markers, and phenotypic variation ranged from 7 to 50%. Among them, four markers Xgwm261-2D, Xgwm495-4B, Xbarc109-4B and Xcfd23-4D were detected under both water regimes. Furthermore, 10 markers were associated with DTC, and four with both parameters PH and DTC at the same plant development stage. The results revealed different allelic effects of associated markers; for example, the 155 bp Xgwm495-4B allele was associated with a reduced height of -11.2 cm under DS and -15.3 cm under WW, whereas the 167 bp allele exhibited increased height effects of 3.9 and 8.1 cm, respectively. This study demonstrates a strong power of joint association analysis and linkage mapping for the identification of important genes in wheat. PMID- 21647606 TI - Enhanced fluorescence anisotropy assay for human cardiac troponin I and T detection. AB - Human cardiac troponin I (hcTnI) and troponin T (hcTnT) are the biomarkers of choice for the diagnosis of cardiac diseases. In an effort to improve assay sensitivity, in this study we developed a novel approach to simultaneously detect hcTnI and hcTnT in homogenous solutions by monitoring enhanced-fluorescence anisotropy changes. Specifically, our design was based on a competition assay by measuring anisotropy change of fluorophore-labeled peptides bound to primary monoclonal antibodies in the presence of nano-gold-modified secondary antibody in response to the presence of target proteins. Enhanced-fluorescence-anisotropy resulted from interaction between the primary antibody and the nano-gold-labeled secondary antibody, which significantly increased the size and decreased tumbling motion of the complex of peptide-antibodies. The measurements were performed to detect hcTnI and hcTnT either individually or simultaneously in a homogenous buffer solution and in the solutions containing human plasma. Our results showed that when fluorescence emission was monitored at a single wavelength selected by a monochromator the assay at all experimental conditions had excellent linear response to the target proteins within the concentration range of 0.5-40 nM. The detection limit is 0.5 nM for both hcTnI and hcTnT in the presence of human plasma. However, when fluorescence emission was monitored using a cutoff filter, the linear response of the assay to the target proteins is within 15-500 pM. The detection limit is 15 pM which is close to the recommended 99th percentile cutoff point for concentrations of hcTnI and hcTnT tests to discriminate healthy and diseased conditions. Homogenous nature, rapid response time, and easy implementation of our assay design make it a useful tool for disease biomarker and protein sensing. PMID- 21647607 TI - Determination of ubiquinone in blood by high-performance liquid chromatography with post-column fluorescence derivatization using 2-cyanoacetamide. AB - It was shown that ubiquinone (CoQ(10)) and ubiquinol (CoQ(10)H(2)) produce fluorescence products under alkaline conditions when reacted with 2 cyanoacetamide. The reaction mixture from CoQ(10) gave fluorescence with excitation and emission maximum wavelengths at 442 nm and 549 nm, respectively. This reaction was considered to proceed via Craven's reaction. Moreover, 2 cyanoacetamide was shown to be a useful reagent for high-performance liquid chromatography (HPLC) with post-column fluorescence derivatization of CoQ(10) and CoQ(10)H(2) in blood. CoQ(10) showed a linear response in the range of 0.32-1276 ng, and the detection limit (S/N = 3) was 0.16 ng. Moreover, the sample pretreatment by deproteinization and extraction of CoQ(10) and CoQ(10)H(2) from plasma using 1-propanol with potassium formate was effective for excellent separation of CoQ(10) and CoQ(10)H(2) from other fluorescent substances in the blood. This simple and rapid pretreatment was considered to minimize the oxidation of CoQ(10)H(2). On the other hand, CoQ(10) and CoQ(10)H(2) in plasma samples obtained by finger prick were detected, as in venous blood obtained by venipuncture. Our method involving the simple and rapid collection of plasma by finger prick and sample pretreatment is thought to be applicable for the determination of CoQ(10)H(2)/total CoQ(10) ratio as a biomarker of oxidative stress. PMID- 21647608 TI - Need for early recognition and therapeutic guidelines of congenital sideroblastic anaemia. AB - We present a patient with iron overload, who was initially diagnosed with hereditary haemochromatosis. Family analysis, however, established that the iron overload was secondary to congenital sideroblastic anaemia. The patient died of a hepatocellular carcinoma, likely a complication of iron overload, despite phlebotomies. Increased awareness, as well as development of evidence-based clinical guidelines, is required for timely diagnosis and adequate treatment. PMID- 21647609 TI - The additive effects of combined murine nuclear migration protein with murine thrombopoietin in vitro and in vivo on normal and myelosuppressed mice. AB - The human homolog of a fungal nuclear migration protein (hNUDC) has recently been shown in in vitro studies to overlap in function with the cytokine factor thrombopoietin (TPO) in its ability to induce human megakaryocyte development. In the present study, we sought to confirm the hypothesis that combination of TPO and NUDC may result in additive or synergistic effects on megakaryocyte maturation in bone marrow. For this purpose, murine bone marrow cells were stimulated with murine NUDC (mNUDC), either alone or in conjunction with murine thrombopoietin (mTPO), in serum-free medium. Studies showed that mNUDC + mTPO treatments resulted in the greatest number of colony-forming unit of megakaryocytes. Concomitantly, mNUDC + mTPO enhanced expression of CD61 and elicited the largest number of megakaryocytes, with higher ploidy and larger size. In addition, in vivo experiments revealed an elevation of platelet levels in normal and thrombocytopenic mice that had been administered mTPO + mNUDC. Moreover, mRNA levels of mNUDC and murine thrombopoietin receptor were much higher than the levels of mTPO mRNA at two different phases of normal murine megakaryocyte maturation. Furthermore, levels of ERK1/2 and Akt phosphorylation are higher with mNUDC in combination with mTPO. This study demonstrates that the combination of mTPO and mNUDC provides additive induction of megakaryocyte maturation in vitro and platelet production in vivo. PMID- 21647610 TI - Backbone and side-chain 1H, 15N, and 13C resonance assignments of Norwalk virus protease. AB - Norovirus protease cleaves the virus-encoded polyprotein into six mature nonstructural proteins, presenting itself as an essential enzyme for the viral replication as well as an attractive target for the antiviral drug development. A deeper understanding of the structural mechanism of the protease substrates/inhibitors interactions by means of solution NMR methods would facilitate a rational design of the virus protease inhibitor. We here report the backbone and side-chain resonance assignment of the protease from Norwalk virus, which is the prototype strain of norovirus. The assignment data has been deposited in the BMRB database under the accession number 17523. PMID- 21647611 TI - Backbone and side chain NMR assignments for the intrinsically disordered cytoplasmic domain of human neuroligin-3. AB - Neuroligins act as heterophilic adhesion molecules at neuronal synapses. Their cytoplasmic domains interact with synaptic scaffolding proteins, and have been shown to be intrinsically disordered. Here we report the backbone and side chain (1)H, (13)C and (15)N resonance assignments for the cytoplasmic domain of human neuroligin 3. PMID- 21647612 TI - Molecular characterization of the modular chitin binding protein Cbp50 from Bacillus thuringiensis serovar konkukian. AB - Bacillus thuringiensis is an insecticidal bacterium whose chitinolytic system may be exploited to improve the insecticidal system of Bt-crops. A nucleotide fragment of 1368 bp from B. thuringiensis serovar konkukian S4, containing the complete coding sequence of the chitin binding protein Cbp50, was cloned and sequenced. Analyses have shown the protein to contain a modular structure consisting of an N-terminal CBM33 domain, two copies of a fibronectin-like domain and a C-terminal chitin binding domain classified as CBM5. The Cbp50 protein was heterologously expressed in Escherichia coli, purified and assessed for chitin binding activity. A deletion mutant (CBD-N; containing only the N-terminal CBM33 domain) of Cbp50 was produced to determine the role of C-terminal domains in the binding activity of the protein. The full-length Cbp50 was shown to bind beta chitin most efficiently followed by alpha-chitin, colloidal chitin and cellulose. The polysaccharide binding activity of CBD-N was drastically decreased. The data demonstrate that both the N-terminal and C-terminal domains of Cbp50 are essential for the efficient binding of chitin. The purified Cbp50 showed antifungal activity against the phytopathogenic fungus Fusarium oxysporum and the opportunistic human pathogen Aspergillus niger. This is the first report of a modular chitin binding protein in bacteria. PMID- 21647613 TI - Molecular cloning, sequence characterization, and tissue expression analysis of chicken sphingomyelin synthase 1 (SMS1). AB - Sphingomyelin synthase 1 (SMS1) is an important cytoplasmic protein which may have functions that go beyond housekeeping function of sphingomyelin (SM) synthesis. In this study, a Hi-Line Brown chicken SMS1 gene was cloned, sequenced, and characterized. The SMS1 full-length coding sequence (CDS) consisted of 1242 nt and encoded 413 amino acids with a molecular weight of 48.54 kDa. The N-terminal sterile alpha motif (SAM) was well conserved between chicken and other animals. The 3D structure of the SMS1 (1-78AA) by homology modeling was similar to that of mouse phosphatidyl ceramide cholinephosphotransferase 1 (2d8c Chain: A (1-97)). The phylogenetic tree analysis revealed that chicken SMS1 had closer genetic relationship with that of land mammals. RT-PCR analysis showed that the SMS1 transcripts were constitutively expressed in 11 tissues tested. Several microRNA target sites were predicted in the CDS of chicken SMS1 mRNA. These data serve as a foundation for further insight into the chicken SMS1 gene. PMID- 21647614 TI - Attenuation of oxidative stress, inflammation and early markers of tumor promotion by caffeic acid in Fe-NTA exposed kidneys of Wistar rats. AB - Iron nitrilotriacetate (Fe-NTA), a chief environmental pollutant, is known for its extensive toxic manifestations on renal system. In the present study, caffeic acid, one of the most frequently occurring phenolic acids in fruits, grains, and dietary supplements was evaluated for its shielding effect against the Fe-NTA induced oxidative, inflammatory, and pathological damage in kidney. Fe-NTA was administered (9 mg Fe/kg body weight) intraperitoneally to the Wistar male rats on 20th day while caffeic acid was administered orally (20 and 40 mg/kg body weight) before administration of Fe-NTA. The intraperitoneal administration of Fe NTA-enhanced lipid peroxidation, xanthine oxidase, and hydrogen peroxide generation with reduction in renal glutathione content, antioxidant enzymes, viz., catalase, glutathione peroxidase, and glutathione reductase. A sharp elevation in the levels of myloperoxidase, blood urea nitrogen (BUN), and serum creatinine has also been observed. Tumor promotion markers viz., ornithine decarboxylase (ODC) and [(3)H] thymidine incorporation into renal DNA were also significantly increased. Treatment of rats orally with caffeic acid (20 and 40 mg/kg body weight) resulted in a significant decrease in xanthine oxidase (P < 0.001), lipid peroxidation (P < 0.001), gamma-glutamyl transpeptidase (P < 0.01), and H(2)O(2) (P < 0.01). There was significant recovery of renal glutathione content (P < 0.001) and antioxidant enzymes (P < 0.001). There was also a reversal in the enhancement of renal ODC activity, DNA synthesis, BUN, and serum creatinine (P < 0.001). All these changes were supported by histological observations. The results indicate that caffeic acid may be beneficial in ameliorating the Fe-NTA-induced oxidative damage and tumor promotion in the kidney of rats. PMID- 21647615 TI - Cholesterol-rich low density lipoproteins are also more oxidized. AB - Cardiovascular diseases are accompanied by active oxygen species and organic free radical generation. The aim of this study was to examine the possibility of using oxidized low-density lipoprotein (oxLDL) as a new diagnostic biomarker. Epidemiological study in populations of Estonia (782 subjects) and Russia (1433 subjects) was carried out in 2007-2009. The screening procedure included standard epidemiological methods. Oxidative stress was assessed by measuring the level of oxLDL using immunoassay method. Positive correlation between the levels of oxLDL and LDL cholesterol was indicated in blood of patients from estonian (r = 0.61; P < 0.05) and russian (r = 0.56; P < 0.05) populations. In russian population oxLDL/HDL cholesterol ratio was higher in the groups with highest risk of atherosclerosis development or manifest coronary artery disease (CAD). Cholesterol-rich low density lipoproteins are also more oxidized. Estimation of oxLDL/HDL ratio may be used as an independent biochemical marker for atherosclerosis. PMID- 21647616 TI - Viral agents causing lower respiratory tract infections in hospitalized children: evaluation of the Speed-Oligo(r) RSV assay for the detection of respiratory syncytial virus. AB - Respiratory syncytial virus (RSV) is the viral agent which is more frequently involved in lower respiratory tract infections (LRTIs) in infants under 1 year of age in developed countries. A new oligochromatographic assay, Speed-Oligo(r) RSV, was designed and optimized for the specific detection and identification of RSV subtypes A and B. The test was evaluated in 289 clinical samples from 169 hospitalized children using an immunochromatography (IC) test, virus isolation by culture, and an in-house real-time polymerase chain reaction (RT-PCR). Other viruses causing LRTIs were investigated by cell culture or PCR-based tests. Sixty two patients were infected by RSV (36.7%). In addition, adenovirus, influenza B, parainfluenza 2, and human metapneumovirus were detected in rates ranging from 5 to 8%. A proportion of 10.1% of the patients had mixed infections. The sensitivity, specificity, and positive and negative predictive values were, respectively, 94.9, 99.4, 98.9, and 97.4% for Speed-Oligo(r) RSV, 92.9, 96.3, 92.9, and 96.3% for RT-PCR/RSV, and 58.4, 98.1, 93.3, and 82.6% for IC. Our rates of viral detection and co-infection were similar to those of previously reported series. Finally, we find that Speed-Oligo(r) RSV is a rapid and easy-to-perform technique for the detection of RSV and the identification of subtypes A and B. PMID- 21647617 TI - Effects of downregulation of aquaporin1 by peptidoglycan and lipopolysaccharide via MAPK pathways in MeT-5A cells. AB - This study was designed to investigate the signaling pathway involved in aquaporin1 (AQP1) expression caused by peptidoglycan (PGN) from Staphylococcus aureus and lipopolysaccharide (LPS) in human pleural mesothelial cell lines (MeT 5A) in vitro. RT-PCR, immunoblot analysis, and immunofluorescence assay were used to determine the relative mRNA and protein levels of AQP1 caused by PGN and LPS in MeT-5A cells. Activation of MAPKs by PGN and LPS was reflected by detecting the phosphorylation constituents of extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun N-terminal kinase (JNK), and p38 using immunoblot. MAPKs inhibitors were used to determine the effects of PGN- and LPS-induced AQP1 expression by immunoblot. AQP1 transcription and protein expression were decreased by PGN and LPS in dose- and time-dependent manners in MeT-5A cells. Both PGN and LPS activated p38/ERK/JNK pathways in MeT-5A cells. Furthermore, downregulation of AQP1 expression by LPS was blocked by SB203580, SP600125, and PD98059, which are inhibitors of p38, JNK, and ERK1/2, respectively. In contrast, downregulation of AQP1 expression by PGN was blocked only by SB203580, not by SP600125 or PD98059, underlying the importance of p38 MAPK in the downregulation of AQP1 expression by PGN in MeT-5A cells. AQP1 expression was decreased by both PGN and LPS in dose- and time-dependent manners in MeT-5A cells. AQP1 expression was down-regulated by PGN via p38 MAPK pathway, while AQP1 expression was down regulated by LPS via p38/JNK/ERK pathways. PMID- 21647618 TI - An extracellular S1-type nuclease of marine fungus Penicillium melinii. AB - An extracellular nuclease was purified 165-fold with a specific activity of 41,250 U/mg poly(U) by chromatography with modified chitosan from the culture of marine fungus Penicillium melinii isolated from colonial ascidium collected near Shikotan Island, Sea of Okhotsk, at a depth of 123 m. The purified nuclease is a monomer with the molecular weight of 35 kDa. The enzyme exhibits maximum activity at pH 3.7 for DNA and RNA. The enzyme is stable until 75 degrees C and in the pH range of 2.5-8.0. The enzyme endonucleolytically degrades ssDNA and RNA by 3'-5' mode to produce 5'-oligonucleotides and 5'-mononucleotides; however, it preferentially degrades poly(U). The enzyme can digest dsDNA in the presence of pregnancy-specific beta-1-glycoprotein-1. The nuclease acts on closed circular double-stranded DNA to produce opened circular DNA and then the linear form DNA by single-strand scission. DNA sequence encoding the marine fungus P. melinii endonuclease revealed homology to S1-type nucleases. The tight correlation found between the extracellular endonuclease activity and the rate of H3-thymidine uptake by actively growing P. melinii cells suggests that this nuclease is required for fulfilling the nucleotide pool of precursors of DNA biosynthesis during the transformation of hyphae into the aerial mycelium and conidia in stressful environmental conditions. PMID- 21647619 TI - Erratum to: When the treatment goal is not cure: are patients informed adequately? PMID- 21647620 TI - Androgens are fundamental in the maintenance of male sexual health. AB - The presence of a sufficient amount of androgens is essential for adequate sexual function in men. Gonadal hormones profoundly affect cognitive functions and, therefore, are fundamental in maintaining sexual desire. The earliest manifestation of hypogonadism is diminished libido. Furthermore, androgens also act peripherally, maintaining the integrity of penile structures and facilitating the erectile mechanisms. Severe androgen deficiency results in anatomical and hemodynamic changes leading to erectile dysfunction that can be corrected with testosterone supplementation. PMID- 21647621 TI - Penile fracture and penile reconstruction. AB - The past decade has seen a significant development in penile reconstruction techniques, and the management of penile fracture has progressively shifted from a conservative approach to early surgical repair. The radial artery-based free flap phalloplasty now represents the gold-standard procedure for total phallic reconstruction both in men and in female-to-male transsexuals. PMID- 21647622 TI - Symptoms suggestive of dumping syndrome after provocation in patients after laparoscopic sleeve gastrectomy. AB - BACKGROUND: Dumping syndrome is a well-known complication after upper gastrointestinal (GI) surgery. There are scarce data in the literature about the incidence of dumping after bariatric operations but, certainly no relation between this syndrome and laparoscopic sleeve gastrectomy (LSG) has been attempted. METHODS: We conducted a prospective clinical study in order to evaluate the potential presence, incidence and severity of Dumping syndrome after LSG. Thirty one non-diabetic morbidly obese patients (eight male, 23 female) eligible for LSG were evaluated. Median age was 38 (22-58 years) and mean body mass index (BMI) was 45.55 (+/- 5.37). The diagnosis of dumping syndrome was based on clinical provocation of signs and symptoms using an oral glucose challenge before and 6 weeks after the operation. The Sigstad's dumping score was estimated in order to separate dumpers from non-dumpers, and the Arts questionnaire was completed to distinguish between early and late dumping. Moreover, blood glucose levels during the oral glucose challenge were measured. RESULTS: No patient had symptoms of dumping after provocation preoperatively, whereas after LSG 9 patients (29%) experienced definite dumping and other 5 patients (16%) symptoms suggestive of dumping syndrome. Arts' questionnaire demonstrated that dumping occurrence after LSG was associated with early symptoms. Late hypoglycaemia occurred in one patient. CONCLUSION: A significant proportion of patients after LSG experienced dumping syndrome upon provocation. It seems that LSG should no longer be considered as a pure restrictive procedure, and it might be an option for heavy sweeters by changing their food tolerance patterns. PMID- 21647623 TI - Health is wealth: methods to improve attendance in a lifestyle intervention for a largely immigrant Filipino-American sample. AB - Diabetes prevention requires lifestyle changes, and traditional educational programs for lifestyle changes have had low attendance rates in ethnic populations. This article describes the development and implementation of an educational program, emphasizing retention strategies, cultural tailoring and community participation. Community-based participatory research approaches were used to adapt and test the feasibility of a culturally tailored lifestyle intervention (named Health is Wealth) for Filipino-American adults at risk for diabetes (n = 40) in order to increase program attendance. A unique feature of this program was the flexibility of scheduling the eight classes, and inclusion of activities, foods and proverbs consistent with Filipino culture. We found that with this approach, overall program attendance for the experimental and wait listed control groups was 88% and participant satisfaction was high with 93% very satisfied. Flexible scheduling, a bilingual facilitator for the classes, and the community-academic partnership contributed to the high attendance for this lifestyle intervention. PMID- 21647624 TI - Comparison of exposures among Arab American and non-Hispanic White female thyroid cancer cases in metropolitan Detroit. AB - Arab American (ArA) women may be at greater risk for thyroid cancer (TC) than White women. This case-case comparison explored differences in known and proposed risk factors of TC among ArA and non-Hispanic White (NHW) female TC cases in metropolitan Detroit. Cases of invasive TC identified from a population-based registry responded to a telephone survey regarding potential TC risk factors. Thirty ArA women (response rate 52%) and 70 NHW women (67%) participated. NHW women reported significantly more prior thyroid disease (TD), family history of TD, hormone use, cumulative years of hormone use, cigarette and alcohol consumption. In adjusted logistic regression analysis, ArA women had significantly higher odds of exposure to dental x-rays (OR = 3.48, CI 1.01-12.00) and medical radiation (OR = 13.58, CI 1.49-124.04) than NHW women. Risk factors for TC may differ among ArA women and their NHW counterparts. PMID- 21647625 TI - A community intervention: AMBER: Arab American breast cancer education and referral program. AB - Although the number of Arab Americans is growing in the United States, there is very little data available on this population's cancer incidence and screening practices. Moreover, there are few interventions addressing their unique needs. This study aims to determine effective strategies for increasing breast cancer screening in at-risk underserved Arab American women. AMBER utilizes a community based participatory approach to conduct formative research and program interventions, including culturally appropriate Arabic language breast cancer education, screening coordination, and cultural competency training for healthcare professionals in New York City. In 2 years, 597 women were educated, 189 underserved women were identified as being in need of assistance, 68 were screened, one new case of breast cancer was detected, and four active cases in need of follow-up reconnected with care. The AMBER model is an important intervention for breast cancer screening and care in the underserved Arab American community. PMID- 21647626 TI - Cognitive and neurochemical alterations in hyperhomocysteinemic rat. AB - Accumulating data have shown that the level of serum homocysteine in patients with mild cognitive impairment, vascular dementia and Alzheimer's disease is higher than normal while the underlying mechanism is not fully understood. Here, a hyperhomocysteinemic rat model was made by maintaining rats on a diet high in methionine. The cognitive behavior, level of monoamine neurotransmitters in brain homogenates and brain-derived neurotrophic factor (BDNF) in cerebral spinal fluid (CSF) were compared between high-methionine diet and control group. The high methionine diet group presented longer mean latency of escape and lesser time in target quadrant in morris maze test, lower level of serotonin and dopamine in cortex homogenates and lower level of BDNF in CSF. Together, our findings provide evidence that hyperhomocysteinemia could cause alterations of monoamine and neurotrophic factor, which might be further pathogenetic mechanisms underlying the cognitive deterioration. PMID- 21647627 TI - Sleep paralysis in narcolepsy: more than just a motor dissociative phenomenon? AB - Sleep paralyses are viewed as pure motor phenomena featured by a dissociated state in which REM-related muscle atonia coexists with a wakefulness state of full consciousness. We present a 59-year-old man diagnosed with narcolepsy experiencing sleep paralysis, who failed to establish the boundaries between real experience and dream mentation during the paralysis: the patient's recall was indeed featured by uncertainty between real/unreal and awaken/dreaming. Hereby, we suggest that sleep paralysis may represent a more complex condition encompassing a dissociated state of mind together with the dissociative motor component. Neurophysiological data (spectral EEG analysis corroborated by cross correlation analysis) reinforce the idea that the patient was in an intermediate state of mind between wake and REM sleep during the paralysis. The persistence of local impaired activity proper of REM sleep in cortical circuits necessary for self-reflective awareness and insight, in conflict with wakefulness-related activation of the remaining brain areas, could account for disrupted processing of afferent inputs in our patient, representing the underlying pathophysiologic substrate for patient's failure to establish the boundaries between real experience and dream mentation. PMID- 21647628 TI - Intractable focal epilepsy contralateral to the side of facial atrophy in Parry Romberg syndrome. AB - We report an unusual case of Parry-Romberg syndrome in which medically refractory focal epilepsy with ongoing epilepsia partialis continua (EPC) arose from the hemisphere contralateral to the side of facial atrophy. Unilateral cerebral involvement was confirmed by multi-modal brain imaging, as well as by electroencephalography (EEG) and magnetoencephalography (MEG). While in many cases of Parry-Romberg syndrome, the side of cerebral involvement is ipsilateral to that of the cutaneous lesion, these "discordant" exceptions imply that other yet undefined mechanisms may be responsible for the distribution of the cutaneous and cerebral pathologies. PMID- 21647629 TI - Early TCD monitoring in brain death: what may be relevant? PMID- 21647630 TI - The appropriate neuroimaging study in persons with epilepsy. AB - This article presents the appropriate neuroimaging for persons with epilepsy (PWE) in resource-limited facilities. PWE from the Epilepsy Clinic, Srinagarind Hospital between November 1, 2003 and January 30, 2005 were enrolled. The inclusion criteria were PWE aged more than 15 years who underwent computed tomography (CT) or magnetic resonance imaging (MRI) of the brain. We compared the abnormal neuroimaging findings by both imaging modalities. A total of 180 out of 370 PWE met the inclusion criteria, comprising 101 men (56.1%) and 79 women (43.9%). There were 75 PWE who underwent only CT imagings, 85 PWE who underwent only MRI and 20 PWE who underwent both CT and MRI studies. CT scan significantly detected brain abnormalities more than MRI in PWE (P = 0.0131). It was also found that CT scan was superior than MRI in detecting stroke and cysticercosis in PWE, whereas MRI was significantly better in the diagnosis of hippocampal sclerosis. Clinicians should be able to choose appropriate brain imaging for PWE, particularly in resource-limited countries. MRI should be preserved for particular brain lesion owing to availability and cost effectiveness. PMID- 21647631 TI - Spatial patterns of TDP-43 neuronal cytoplasmic inclusions (NCI) in fifteen cases of frontotemporal lobar degeneration with TDP-43 proteinopathy (FTLD-TDP). AB - Neuronal cytoplasmic inclusions (NCI) immunoreactive for transactive response DNA binding protein (TDP-43) are the pathological hallmark of frontotemporal lobar degeneration with TDP-43 proteinopathy (FTLD-TDP). We studied the spatial patterns of the TDP-43 immunoreactive NCI in the frontal and temporal cortex of 15 cases of FTLD-TDP. The NCI were distributed parallel to the tissue boundary predominantly in regular clusters 50-400 MUm in diameter. In five cortical areas, the size of the clusters approximated to the cells of the cortico-cortical pathways. In most regions, cluster size was smaller than 400 MUm. There were no significant differences in spatial patterns between familial and sporadic cases. Cluster size of the NCI was not correlated with disease duration, brain weight, Braak stage, or disease subtype. The spatial pattern of the NCI was similar to that of neuronal inclusions in other neurodegenerative diseases and may reflect a common pattern of degeneration involving the cortico-cortical projections. PMID- 21647632 TI - MELAS/SANDO overlap syndrome associated with POLG1 mutations. PMID- 21647633 TI - HIF-1alpha ODD polypeptides increased the expression of HIF1 and VEGF in hypoxic rat cortical neuron. AB - To investigate the effect of HIF-1alpha ODD polypeptides on the expression of HIF 1alpha and VEGF in primary rat cortical neuron in hypoxia. Primary cultured neurons were exposed to hypoxia for 2, 4, and 8 h. Tat-ODD fusion polypeptide was added before hypoxia in the experimental groups. The expression of HIF-1alpha and VEGF was detected by Western blot and real time PCR. The levels of HIF-1alpha and VEGF peaked at 4 h then declined in hypoxia. The expression of HIF-1alpha and VEGF was increased by tat-ODD fusion polypeptide. tat-ODD fusion polypeptide might improve HIF1 adaptation to hypoxia by increased expression of HIF-1alpha and VEGF. PMID- 21647634 TI - Monoclonal antibody to six transmembrane epithelial antigen of prostate-4 influences insulin sensitivity by attenuating phosphorylation of P13K (P85) and Akt: possible mitochondrial mechanism. AB - We examined the effects of anti-six-transmembrane epithelial antigen of the prostate-4 (STEAP4) antibodies on glucose transport in mature adipocytes and determined the mechanism of insulin resistance in obesity. Western blotting was performed to determine STEAP4 expression, to assess translocation of insulin sensitive glucose transporter 4 (GLUT4), and to measure phosphorylation and total protein content of insulin-signaling proteins. Confocal laser microscopy and flow cytometry were used to detect intracellular reactive oxygen species (ROS) and fluctuations in mitochondrial membrane potential (DeltaPsi). ATP production was measured by using a luciferase-based luminescence assay kit. After the application of anti-STEAP4 antibodies at 0.002 mg/mL, adipocytes exhibited reduced insulin-stimulated glucose transport by attenuating the phosphorylation of IRS-1, PI3K (p85), and Akt. The antibodies also potentially increase the level of ROS and decrease cellular ATP production and DeltaPsi. In conclusion, (i) STEAP4 regulates the function of IRS-1, PI3K, and Akt and decreases insulin induced GLUT4 translocation and glucose uptake; (ii) ROS-related mitochondrial dysfunction may be related to a reduced IRS-1 correlation with the PI3K signaling pathway, leading to insulin resistance. These observations highlight the potential role of STEAP4 in glucose homeostasis and possibly in the pathophysiology of type 2 diabetes related to obesity and may provide new insights into the mechanisms of insulin resistance in obesity. PMID- 21647635 TI - A role for anions in ATP synthesis and its molecular mechanistic interpretation. AB - ATP, the 'universal biological energy currency', is synthesized by utilizing energy either from oxidation of fuels or from light, via the process of oxidative and photo-phosphorylation respectively. The process is mediated by the enzyme F(1)F(0)-ATP synthase, using the free energy of ion gradients in the final energy catalyzing step, i.e., the synthesis of ATP from ADP and inorganic phosphate (P(i)). The details of the molecular mechanism of ATP synthesis are among the most important fundamental issues in biology and hence need to be properly understood. In this work, a role for anions in making ATP has been found. New experimental data has been reported on the inhibition of ATP synthesis at nanomolar concentrations by the potent, specific anion channel blockers 4,4' diisothiocyanostilbene-2, 2'-disulphonic acid (DIDS) and tributyltin chloride (TBTCl). Based on these inhibition studies, attention has been drawn to anion translocation (in addition to proton translocation) as a requirement for ATP synthesis. The type of inhibition has been quantified and an overall kinetic scheme for mixed inhibition that explains the data has been evolved. The experimental data and the type of inhibition found have been interpreted in the light of the torsional mechanism of energy transduction and ATP synthesis (Nath J Bioenerg Biomembr 42:293-300, 2010a; J Bioenerg Biomembr 42:301-309, 2010b). This detailed and unified mechanism resolves long-standing problems and inconsistencies in the first theories (Slater Nature 172:975-978, 1953; Williams J Theor Biol 1:1-17, 1961; Mitchell Nature 191:144-148, 1961; Mitchell Biol Rev 41:445-502, 1966), makes several novel predictions that are experimentally verifiable (Nath Biophys J 90:8-21, 2006a; Process Biochem 41:2218-2235, 2006b), and provides us with a new and fruitful paradigm in bioenergetics. The interpretation presented here provides intelligent answers to the unexplained existing results in the literature. It is shown that mechanistic interpretation of the experimental data requires substantial addition to available conceptual foundations such that present concepts, theories, and mechanisms must be revised. PMID- 21647637 TI - Identification of functional BrFAD2-1 gene encoding microsomal delta-12 fatty acid desaturase from Brassica rapa and development of Brassica napus containing high oleic acid contents. AB - Microsomal delta-12 fatty acid desaturase (FAD2) functions in the first committed step of the biosynthesis of polyunsaturated fatty acids via the desaturation of oleic acid to linoleic acid. In this study, two FAD2 genes were identified through genome-wide analysis of Brassica rapa. One BrFAD2-1 gene harbors functional sequence information, but another BrFAD2-2 gene has mutations that generated a premature stop codon, rendering it nonfunctional. From a database of 120,000 B. rapa expressed sequence tags, we determined that all sequences coding for FAD2 corresponded to the BrFAD2-1 gene. The BrFAD2-1 protein was shown to share high sequence homology (71-99%) with FAD2 proteins from other plant species. An intron in the 5'-untranslated region and three histidine boxes in the protein, which are characteristic of plant FAD2 genes, have been well-conserved. BrFAD2-1 transcripts were detected in various organs of B. rapa. When a pBrFAD2 1:mRFP construct was introduced into tobacco epidermal cells, the fluorescent signal was noted in the endoplasmic reticulum. Ectopic expression of BrFAD2 1:mRFP complemented the Arabidopsis fad2-2 mutant. Finally, transgenic Korean rapeseed Tammi containing high oleic acid contents (78 mol%) was developed via the expression of the BrFAD2-1 gene in an antisense orientation. The data demonstrate that B. rapa harbors only one functional FAD2 that can be utilized for the development of the high-oleic acid Korean rapeseed cultivar Tammi, which might be useful for both human consumption and industrial applications. PMID- 21647638 TI - An insertional mutagenesis programme with an enhancer trap for the identification and tagging of genes involved in abiotic stress tolerance in the tomato wild related species Solanum pennellii. AB - Salinity and drought have a huge impact on agriculture since there are few areas free of these abiotic stresses and the problem continues to increase. In tomato, the most important horticultural crop worldwide, there are accessions of wild related species with a high degree of tolerance to salinity and drought. Thus, the finding of insertional mutants with other tolerance levels could lead to the identification and tagging of key genes responsible for abiotic stress tolerance. To this end, we are performing an insertional mutagenesis programme with an enhancer trap in the tomato wild-related species Solanum pennellii. First, we developed an efficient transformation method which has allowed us to generate more than 2,000 T-DNA lines. Next, the collection of S. pennelli T(0) lines has been screened in saline or drought conditions and several presumptive mutants have been selected for their salt and drought sensitivity. Moreover, T-DNA lines with expression of the reporter uidA gene in specific organs, such as vascular bundles, trichomes and stomata, which may play key roles in processes related to abiotic stress tolerance, have been identified. Finally, the growth of T-DNA lines in control conditions allowed us the identification of different development mutants. Taking into account that progenies from the lines are being obtained and that the collection of T-DNA lines is going to enlarge progressively due to the high transformation efficiency achieved, there are great possibilities for identifying key genes involved in different tolerance mechanisms to salinity and drought. PMID- 21647639 TI - IVM is an alternative for patients with PCO after failed conventional IVF attempt. AB - PURPOSE: To determine if IVM of oocytes from unstimulated cycle is a treatment option for patients who did not deliver after standard IVF-ET. METHOD: Twenty three women with PCO, thirteen of them with normal cycles and all <35 years old, who failed IVF served as their own control. During the control IVF cycle patients were stimulated with 1730.7 +/- 639.5 IU recombinant FSH, a long Buserelin acetate protocol was used and embryo transfer was performed on day 2 or 3 after ICSI. After failed IVF immature oocytes were aspirated transvaginally from antral follicles during spontaneous menstrual cycle. Embryo transfer was performed 2 or 3 days later. RESULT: 11.4 +/- 4.8 mature oocytes and 6.7 +/- 3.2 embryos were produced with IVF, which served as the control, compared to 9.7 +/- 4.5 mature oocytes and 6.2 +/- 3.2 embryos with IVM. There was one clinical pregnancy in the IVF group which did not result in a live birth where as five singleton and one pair of twins with healthy live births and one miscarriage in the IVM group. CONCLUSION: IVM does not involve ovarian stimulation with possible financial and health consequences. It may be an useful treatment after unsuccessful IVF. PMID- 21647640 TI - Differentially expressed micoRNAs in human oocytes. AB - PURPOSE: To identify differentially expressed microRNAs (miRNAs) and expression patterns of specific miRNAs during meiosis in human oocytes. MATERIALS AND METHODS: To identify differentially expressed miRNAs, GV oocytes and MII oocytes matured at conventional FSH levels (5.5 ng/ml) were analyzed by miRNA microarray. Real-time RT-PCR was used to confirm the changed miRNAs. To validate the dynamic changes of miRNAs from GV to MII stages, oocytes were divided into four groups (#1-4), corresponding to GV oocytes, MI oocytes, MII oocytes matured in conventional FSH level and MII oocytes matured in high FSH level (2,000 ng/ml) respectively. RESULTS: Compared with GV oocytes, MII oocytes exhibited up regulation of 4 miRNAs (hsa-miR-193a-5p, hsa-miR-297, hsa-miR-625 and hsa-miR 602), and down-regulation of 11 miRNAs (hsa-miR-888*, hsa-miR-212, hsa-miR-662, hsa-miR-299-5p, hsa-miR-339-5p, hsa-miR-20a, hsa-miR-486-5p, hsa-miR-141*, hsa miR-768-5p, hsa-miR-376a and hsa-miR-15a). RT-PCR analysis of hsa-miR-15a and hsa miR-20a expression revealed concordant dynamic changes in oocytes from group 1 to group 4. CONCLUSION(S): Specific miRNAs in human oocytes had dynamic changes during meiosis. High-concentration FSH in IVM medium led to reverse effect on the expression of hsa-miR-15a and hsa-miR-20a. PMID- 21647641 TI - Feasibility of multiple short, 40-s, intra-procedural ECG recordings to detect immediate changes in heart rate variability during catheter ablation for arrhythmias. AB - PURPOSE: This study aims to evaluate a method to detect heart rate variability (HRV) changes using short ECG segments during ablation for arrhythmias. METHODS: HRV was averaged from sequentially shorter time windows from 5-min ECG recordings in 15 healthy volunteers. The 40-s window was identified as the shortest duration that yielded reproducible values in high frequency (HF) and low frequency (LF) HRV. This method was validated in patients undergoing tilt table testing to see if the expected modulation in HRV that occurs prior to syncope could be detected from multiple 40-s recordings. Lastly, this method was used to assess HRV changes in 75 patients undergoing ablation for atrial fibrillation (AF) and other arrhythmias, to see if autonomic modulation as a result of ablation could be detected. A further 14 patients had stepwise HRV measurements at different stages of the AF ablation procedure to determine whether intra-procedural HRV changes could be detected. RESULTS: HRV, averaged from multiple 40-s recordings, demonstrated the expected increase immediately preceding syncope compared with baseline (LF: 341 +/- 311-1,536 +/- 1,368 ms(2), p< 0.05; HF: 342 +/- 339-1,628 +/- 1,755 ms(2), p < 0.05). AF ablation, particularly following right pulmonary vein circumferential ablation, produced immediately detectable reductions in LF (153 +/- 251-50 + 116 ms(2), p < 0.001) and HF (86 +/- 195-33 +/- 83 ms(2), p < 0.001) without any change in RR interval (877 +/- 191-843 +/- 220 ms, p = 0.261). Ablation for atrial flutter did not change the mean RR interval, LF or HF HRV. CONCLUSION: Averaging multiple 40-s windows give valid HF and LF HRV measurements that enable detection of intra-procedural changes. Left atrial ablation around the right-sided pulmonary veins is unique in producing reductions in HRV. This method has the potential for use as an endpoint marker for adjunctive autonomic ablation procedures. PMID- 21647642 TI - Advances in syncope: with emphasis on reflex syncope. AB - Syncope is a sudden transient loss of consciousness associated with loss of postural tone, caused by bradycardia, tachycardia, or abnormal autonomic reflexes. Pacing and implantable defibrillators treat but do not cure arrhythmias. Reflex syncope has remained a therapeutic challenge. Remarkably, recent advances in the treatment of the reflex syncopal syndromes are (1) the recognition that many pharmacologic and device-based treatments do not really work, (2) complex subdivisions among the reflex syncope types can be simplified, and (3) effective therapies are often common sense applications of behavioral modifications and physical maneuvers. Many of the treatments have been available for decades and have been supplemented by a renewed interest in the physiologic approach while maintaining a role for evidence-based pharmacologic and device based therapy. Streamlined workups in dedicated syncope centers may be effective, both economically and therapeutically. In the long term, important advances in syncope are likely to come from the still embryonic field of genetics. Beyond genetic counseling, the greatest benefits lie in the future. PMID- 21647643 TI - Duration of the A(H)-A(Md) interval predicts occurrence of AV-block after radiofrequency ablation of the slow pathway. AB - PURPOSE: Modification of the slow pathway (SP) of the atrio-ventricular node by radiofrequency ablation is the most effective treatment to cure AV nodal reentry tachycardia (AVNRT). However, this therapy may be complicated by AV-block (AVB). We sought to evaluate the predictive value of the A(H)-A(Md) interval-the electrical delay between atrial signals on the His- and the ablation-catheter upon development of AVB during SP ablation. METHODS: The associations between A(H)-A(Md) interval, occurrence of ventriculo-atrial block (VAB) during junctional activity (JA) and transient or permanent AVB were analyzed retrospectively for 1585 RF applications at the SP in 393 patients diagnosed with AVNRT. The value of A(H)-A(Md) was further tested prospectively in 118 AVNRT patients, who were only ablated at targets with intervals >20 ms. RESULTS: Forty six RF deliveries resulted in transient or permanent AV-conduction disturbances. Shorter A(H)-A(Md) intervals were associated with the occurrence of VAB during JA (p < 0.001) and AVB (p < 0.001). A(H)-A(Md) was the strongest predictor for VAB or AVB in multivariate regression analyses, followed by the radiological distance between the catheters. In the prospective study, permanent high-degree AVB was not observed when the A(H)-A(Md) at the ablation site was >20 ms. CONCLUSION: The A(H)-A(Md) interval is a better predictor for occurrence of conduction block during ablation for AVNRT than the radiological distance between the His- and the ablation-catheter. The risk of permanent AVB can be minimized, if only sites with an A(H)-A(Md) longer than 20 ms are targeted for ablation. PMID- 21647644 TI - To publish or not to publish....... PMID- 21647645 TI - Optimizing treatment strategies in paediatric atypical hemolytic uremic syndrome. PMID- 21647646 TI - Long-term renal function under plasma exchange in atypical hemolytic uremic syndrome. PMID- 21647647 TI - Species difference in the timing of gaze movement between chimpanzees and humans. AB - How do humans and their closest relatives, chimpanzees, differ in their fundamental abilities for seeing the visual world? In this study, we directly compared the gaze movements of humans and the closest species, chimpanzees, using an eye-tracking system. During free viewing of a naturalistic scene, chimpanzees made more fixations per second (up to four) than did humans (up to three). This species difference was independent of the semantic variability of the presented scenes. The gap-overlap paradigm revealed that, rather than resulting from the sensitivity to the peripherally presented stimuli per se, the species difference reflected the particular strategy each species employed to solve the rivalry between central (fixated) and peripheral stimuli in their visual fields. Finally, when presented with a movie in which small images successively appeared/disappeared at random positions at the chosen presentation rate, chimpanzees tracked those images at the point of fixation for a longer time than did humans, outperforming humans in their speed of scanning. Our results suggest that chimpanzees and humans differ quantitatively in their visual strategies involving the timing of gaze movement. We discuss the functional reasons for each species' employing such specific strategies. PMID- 21647648 TI - Chimpanzee problem-solving: contrasting the use of causal and arbitrary cues. AB - Humans are able to benefit from a causally structured problem-solving context rather than arbitrarily structured situations. In order to better understand nonhuman causal cognition, it is therefore important to isolate crucial factors that might differentiate between events that follow a purely spatial and temporal contingency and those that hold a "true" causal relationship. In the first of two experiments, chimpanzee subjects were required to detect a bottle containing juice from five opaque bottles of equal shape and size. In the causal condition, the juice bottle looked identical to the other four bottles, only it was much heavier than the others. In the arbitrary condition, the weight of all five bottles was identical, but the juice bottle was color-marked differently. Since bottle opening was made difficult (and therefore costly), the question was whether subject's manipulative behavior would be random or somehow influenced by the nature of the provided information. Our results show that subjects detected and opened the juice bottle significantly faster when weight was the discriminating feature (causal condition) compared to situations in which the discrimination was necessarily based on a color-cue (arbitrary condition). Experiment 2 ruled out the possibility of a general learning bias toward tactile rather than visual information in chimpanzees. When tested in a simple exchange paradigm that prevented any use of causal information, no predominance of a tactile cue (weight) over a visual cue (color) could be found. Furthermore--and in contrast to the causal condition in Experiment 1--no learning occurred during the course of Experiment 2, neither in the weight nor in the color condition. We therefore conclude that chimpanzees can more easily determine the content of an object based on its causal properties compared to situations in which the only available information is a pure arbitrary regularity. This supports the view that chimpanzees' causal cognition does not rely on mere perceptual information but also on structural abstraction about their physical environment. PMID- 21647649 TI - Pigeons discriminate between human feeders. AB - Considered as plague in many cities, pigeons in urban areas live close to human activities and exploit this proximity to find food which is often directly delivered by people. In this study, we explored the capacity of feral pigeons to take advantage of this human-based food resource and discriminate between friendly and hostile people. Our study was conducted in an urban park. Pigeons were fed by two experimenters of approximately the same age and skin colour but wearing coats of different colours. During the training sessions, the two human feeders displayed different attitudes: one of the feeders was neutral and the second was hostile and chased away the pigeons. During the two test phases subsequent to the training phase, both feeders became neutral. Two experiments were conducted, one with one male and one female feeder and the second with two female feeders. In both experiments, the pigeons learned to quickly (six to nine sessions) discriminate between the feeders and maintained this discrimination during the test phases. The pigeons avoided the hostile feeder even when the two feeders exchanged their coats, suggesting that they used stable individual characteristics to differentiate between the experimenter feeders. Thus, pigeons are able to learn quickly from their interactions with human feeders and use this knowledge to maximize the profitability of the urban environment. This study provides the first experimental evidence in feral pigeons for this level of human discrimination. PMID- 21647650 TI - Federal disaster mental health response and compliance with best practices. AB - This study investigated the comprehensiveness of disaster mental health state plans and their adherence to published best practices in three states that experienced post-9/11 federally-declared disasters. There were 59 disaster mental health best practices used in this study to assess each state disaster mental plan's compliance with best practices; the states demonstrated a range of adherence to the best practices. This research may serve as a guide for those developing disaster mental health plans and encourage further considerations in disaster mental health response. PMID- 21647652 TI - The impact of post-infectious functional gastrointestinal disorders and symptoms on the health-related quality of life of US military personnel returning from deployment to the Middle East. AB - BACKGROUND AND AIM: Mental and physical health-related quality of life (HRQOL) are important health impact measures following military deployment. While conditions such as post-traumatic stress disorder (PTSD) are known to adversely affect QOL, little is known about the effect of post-infectious functional gastrointestinal disorders (PI-FGID). Our aim was to evaluate the risk of PI-FGID and its impact on HRQOL among military personnel returning from deployment. METHODS: A cross-sectional cohort of active-duty military deployed to Egypt or Turkey between 2004 and 2005 was asked to complete a questionnaire (Rome II and SF-36 instruments) on travelers' diarrhea (TD) during deployment and FGID symptoms and HRQOL 6 months after returning from deployment. RESULTS: A total of 121 military personnel returning from Egypt (n = 33) and Turkey (n = 88) completed the post-deployment questionnaire. Nearly half (48.3%) met the definition for an FGID at the time of the survey, and 53% of individuals reporting one or more episodes of TD during deployment developed an FGID, compared to 33% of those not reporting TD (odds ratio [OR] 2.2, P = 0.08). Compared to those not meeting the FGID criteria, those with post-deployment FGID had lower mean mental HRQOL scores (-13.4%, P < 0.0001) and lower physical HRQOL scores (-7.2%, P = 0.004). CONCLUSIONS: There was a high prevalence of FGID symptoms in military personnel returning from deployment, and TD was a noted risk factor. FGID and symptoms decreased QOL, with mental HRQOL being affected more than physical HRQOL. These findings require further research in order to assess the long-term impact of these and other post-infectious sequela related to TD during deployments among returning veterans. PMID- 21647653 TI - Hepatobiliary disease associated with hypereosinophilia: a report of two patients and a review of the literature. PMID- 21647651 TI - Cryptogenic chronic hepatitis and its changing guise in adults. AB - Cryptogenic chronic hepatitis is a disease that is unexplained by conventional clinical, laboratory and histological findings, and it can progress to cirrhosis, develop hepatocellular carcinoma, and require liver transplantation. The goals of this review are to describe the changing phenotype of cryptogenic chronic hepatitis in adults, develop a diagnostic algorithm appropriate to current practice, and suggest treatment options. The frequency of cryptogenic hepatitis is estimated at 5.4%. Cryptogenic cirrhosis is diagnosed in 5-30% of patients with cirrhosis, and it is present in 3-14% of adults awaiting liver transplantation. Nonalcoholic fatty liver disease has been implicated in 21-63% of patients, and autoimmune hepatitis is a likely diagnosis in 10-54% of individuals. Viral infections, hereditary liver diseases, celiac disease, and unsuspected alcohol or drug-induced liver injury are recognized infrequently in the current cryptogenic population. Manifestations of the metabolic syndrome heighten the suspicion of nonalcoholic fatty liver disease, and the absence of hepatic steatosis does not discount this possibility. The diagnostic scoring system of the International Autoimmune Hepatitis Group can support the diagnosis of autoimmune hepatitis in some patients. Certain genetic mutations may have disease-specificity, and they suggest that some patients may have an independent and uncharacterized disease. Corticosteroid therapy is effective in patients with autoimmune features, and life-style changes and specific therapies for manifestations of the metabolic syndrome are appropriate for all obese patients. The 1- and 5-year survivals after liver transplantation have ranged from 72-85% to 58-73%, respectively. PMID- 21647654 TI - Double-blind randomized controlled trial of rifaximin for persistent symptoms in patients with celiac disease. AB - BACKGROUND: Small intestinal bacterial overgrowth (SIBO) is one cause of a poor response to a gluten-free diet (GFD) and persistent symptoms in celiac disease. Rifaximin has been reported to improve symptoms in non-controlled trials. AIMS: To determine the effect of rifaximin on gastrointestinal symptoms and lactulose hydrogen breath tests in patients with poorly responsive celiac disease. METHODS: A single-center, double-blind, randomized, controlled trial of patients with biopsy-proven celiac disease and persistent gastrointestinal symptoms despite a GFD was conducted. Patients were randomized to placebo (n = 25) or rifaximin (n = 25) 1,200 mg daily for 10 days. They completed the Gastrointestinal Symptom Rating Scale (GSRS) and underwent lactulose-hydrogen breath tests at weeks 0, 2, and 12. An abnormal breath test was defined as: (1) a rise in hydrogen of >=20 parts per million (ppm) within 100 min, or (2) two peaks >=20 ppm over baseline. RESULTS: GSRS scores were unaffected by treatment with rifaximin, regardless of baseline breath tests. In a multivariable regression model, the duration of patients' gastrointestinal symptoms significantly predicted their overall GSRS scores (estimate 0.029, p < 0.006). According to criteria 1 and 2, respectively, SIBO was present in 55 and 8% of patients at baseline, intermittently present in 28 and 20% given placebo, and 28 and 12% given rifaximin. There was no difference in the prevalence of SIBO between placebo and treatment groups at weeks 2 and 12. CONCLUSIONS: Rifaximin does not improve patients' reporting of gastrointestinal symptoms and hydrogen breath tests do not reliably identify who will respond to antibiotic therapy. PMID- 21647655 TI - Enhanced expression of CXCL13 in human Helicobacter pylori-associated gastritis. AB - BACKGROUND AND AIMS: Chemokine CXC ligand 13 (CXCL13) and CXC receptor type 5 (CXCR5) are constitutively expressed in tertiary lymphoid follicles where the CXCL13/CXCR5 system regulates B lymphocytes homing. In this study, we sought to examine CXCL13 expression in the H. pylori-infected and -uninfected gastric mucosa and to elucidate the implication in the pathogenesis of HAG in humans. METHODS: Using endoscopic biopsies taken from the gastric antrum of 29 subjects infected with Helicobacter pylori and 22 uninfected subjects, mucosal CXCL13 mRNA and protein levels were measured by real-time polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. RESULTS: The CXCL13 expression levels were significantly more elevated in H. pylori-positive patients than uninfected ones. The CXCL13 expression levels correlated with the degree of chronic gastritis and bacterial colonization. Immunohistochemistry and in vitro infection assay showed that CXCL13 was not produced by the gastric epithelium, but the alpha-smooth muscle antigen expressing mesenchymal cells were the possible source of CXCL13 within H. pylori-infected gastric mucosa. CXCR5 immunostaining was seen in the CD20-positive lymphoid aggregates. CONCLUSIONS: The enhanced induction of CXCL13 may be involved in the pathogenesis of H. pylori associated gastritis. PMID- 21647656 TI - Acute effects of Ginkgo biloba extract on vascular function and blood pressure. AB - We investigated whether a single dose of standardized Ginkgo biloba extract (GBE) can improve vascular function. A randomised controlled crossover trial was conducted on 14 young healthy men, who received GBE or placebo. The digital volume pulse was monitored to measure reflection index (DVP-RI) and stiffness index (DVP-SI) and peripheral augmentation index (pAIx) was assessed using radial pulse wave analysis at baseline and 2, 4 and 6 h after treatment. DVP-SI was slightly higher 2 h following GBE compared to placebo (P < 0.05); other outcome variables were unaffected by treatment. PMID- 21647657 TI - The effect of gender on force, muscle activity, and frontal plane knee alignment during maximum eccentric leg-press exercise. AB - PURPOSE: To investigate for gender differences during eccentric leg-press exercise. Tears of the anterior cruciate ligament (ACL) are considered to be related to eccentric tasks, altered neuromuscular control (e.g., reduced co contraction of hamstrings), and increased knee abduction (valgus alignment). Based on these observations and the fact that ACL tears are more common in women, it was hypothesized that men and women differ significantly with regard to key parameters of force, knee stabilization, and muscle activity when exposed to maximum eccentric leg extension. METHODS: Thirteen women and thirteen men were matched for age and physical activity. They performed maximum isokinetic eccentric leg-pressing against footplates of varied stability. The latter was done because earlier studies had shown that perturbational test conditions might be relevant in respect of ACL injuries. Key parameters of force, frontal plane knee stabilization, and muscle recruitment of significant muscles crossing the knee were recorded. RESULTS: The 'force stabilization deficit' (difference between maximum forces under normal and perturbed leg-pressing) did not differ significantly between genders. Likewise, parameters of muscle activity and frontal plane leg stabilization revealed no significant differences between men and women. CONCLUSION: This study is novel, in that gender differences in parameters of force, muscle activity, and leg kinematic were investigated during functional conditions of eccentric leg-pressing. No gender differences were observed in the measured parameters. However, the conclusion should be viewed with caution because the findings concurred with, but also contrasted, previous research in this field. LEVEL OF EVIDENCE: Diagnostic study, Level III. PMID- 21647658 TI - Bupivacaine, ropivacaine, and morphine: comparison of toxicity on human hamstring derived stem/progenitor cells. AB - PURPOSE: Bupivacaine, ropivacaine, and morphine are commonly administered intraarticularly after anterior cruciate ligament (ACL) reconstruction. However, their effects on human tendon stem/progenitor cells (TSPC) have not been studied. Therefore, this study investigates the cytotoxicity of these analgetics on TSPC. METHODS: Cells were isolated from human hamstring grafts of 3 female (age 15, 16 and 59) and 2 male patients (age 16 and 47). Cells were incubated using 0.5% bupivacaine, 0.5/0.75% ropivacaine, and 0.025% morphine. Cell viability was assessed after 0.5, 2, and 6 h using live/dead assay. Metabolic activity and apoptosis were measured by WST- and Annexin-V-FACS-assay after 2 h. RESULTS: Cell viability remained unchanged after 0.5 h in all groups, while treatment with bupivacaine and 0.5/0.75% ropivacaine resulted in a complete cell loss after 6 h. Contrarily, morphine showed no cytotoxic effect. Cell viability and metabolism were significantly reduced after treatment with bupivacaine (22.1; 8.3%) and 0.75% ropivacaine (56.5; 23.8%), while 0.5% ropivacaine and morphine showed no significant difference compared with controls. Apoptosis was significantly induced after incubation with bupivacaine (58.1%) and 0.75% ropivacaine (26.2%), whereas 0.5% ropivacaine only led to a slight induction compared with morphine and controls. CONCLUSIONS: Clinically administered concentrations of bupivacaine (0.5%) and ropivacaine (0.75%) have a significant cytotoxic effect on human TSPC in vitro, while ropivacaine in a concentration of 0.5% has a mild but not significant effect on apoptosis and cell metabolism. In contrast, morphine does not affect cell survival, metabolism, or apoptosis. Knowing that morphine provides comparable to even prolonged pain reduction after ACL reconstruction, the presented in vitro study suggests morphine as a potentially less toxic analgetic drug for intraarticular application in clinical practice. PMID- 21647659 TI - Necrotizing fasciitis of the knee following primary total knee arthroplasty. AB - Necrotizing fasciitis is a rare, life-threatening and rapidly spreading soft tissue infection that results in necrosis of the muscle, fascia and surrounding tissue. It can be result of a polymicrobial synergistic infection or a streptococcal infection. The authors report a case of necrotizing fasciitis occurring in the knee of a 65-year-old woman following an uneventful primary total knee arthroplasty and resulting in above-the-knee amputation. Having in mind severe infections like necrotising fasciitis, one should be aware of the possibility of such postoperative complications especially in patients with risk factors even in routine procedures like a total knee arthroplasty. PMID- 21647660 TI - Can femoroacetabular impingement and hip dysplasia be distinguished by clinical presentation and patient history? AB - PURPOSE: Hip dysplasia and femoroacetabular impingement may both lead to disability and hip osteoarthritis. The purpose of the current study was to compare the two entities in order to define differences in their clinical presentation. METHODS: The clinical presentation and WOMAC items, subscales and overall sum score were compared between 37 patients with femoroacetabular impingement and 37 patients with hip dysplasia. RESULTS: The average duration of symptoms was 33.3 +/- 31.6 months in patients with femoroacetabular impingement and 34.5 +/- 39.0 months in patients with dysplasia (p = 0.885). The anterosuperior impingement test was positive in all patients with femoroacetabular impingement and in 92% of hip dysplasia patients (p = 0.061). Mean internal rotation and abduction was significantly less in patients with femoroacetabular impingement (p = 0.001 and 0.007). The WOMAC subscales for pain, stiffness, and functionality as well as the overall sum score were not significantly different between patients with femoroacetabular impingement and patients with hip dysplasia. The qualitative analysis of WOMAC items revealed that symptoms related to sitting were significantly more often rated to be present in femoroacetabular impingement than in hip dysplasia patients. In the quantitative analysis, only the item 'getting in or out of a car' was rated significantly more severe in patients with femoroacetabular impingement. CONCLUSION: In addition to obtaining a detailed history and examination, radiographic studies are needed for differentiation of the two entities and for the decision on treatment strategy. PMID- 21647661 TI - Olfactory sensitivity for six amino acids: a comparative study in CD-1 mice and spider monkeys. AB - Using a conditioning paradigm, the olfactory sensitivity of five CD-1 mice for the L- and D-forms of cysteine, methionine, and proline was investigated. With all six stimuli, the animals discriminated concentrations <= 0.1 ppm (parts per million) from the odorless solvent, and with three of the six stimuli the best scoring animals were even able to detect concentrations <0.1 ppb (parts per billion). Three spider monkeys tested in parallel were found to detect the same six stimuli at concentrations <1 ppm, and with four of the six stimuli the best scoring animals detected concentrations <= 1 ppb. Both CD-1 mice and spider monkeys displayed a higher olfactory sensitivity with the L- and D-forms of cysteine and methionine than with the prolines, suggesting an important role of the sulfur-containing functional groups for detectability. Accordingly, the across-odorant patterns of detection thresholds obtained with mice and spider monkeys showed a significant positive correlation. A comparison of the detection thresholds between the two species tested here and those obtained in human subjects suggests that neither the number of functional olfactory receptor genes nor the absolute or the relative size of the olfactory bulbs reliably predicts a species' olfactory sensitivity for amino acids. PMID- 21647662 TI - Pharmacokinetics and cerebral distribution of glycine administered to rats. AB - High doses of glycine have been reported to improve negative schizophrenic symptoms, suggesting that ingested glycine activates glutamatergic transmission via N-methyl-D-aspartate (NMDA) receptors. However, the pharmacokinetics of administered glycine in the brain has not been evaluated. In the present study, the time- and dose-dependent distributions of administered glycine were investigated from a pharmacokinetic viewpoint. Whole-body autoradiography of radiolabeled glycine was performed, and time-concentration curves for glycine and serine in plasma, cerebrospinal fluid (CSF), and brain tissues were obtained. Furthermore, pharmacokinetic parameters were calculated. For a more detailed analysis, the amount of glycine uptake in the brain was evaluated using the brain uptake index method. Radiolabeled glycine was distributed among periventricular organs in the brain. Oral administration of 2 g/kg of glycine significantly elevated the CSF glycine concentration above the ED50 value for NMDA receptors. The glycine levels in CSF were 100 times lower than those in plasma. Glycine levels were elevated in brain tissue, but with a slower time-course than in CSF. Serine, a major metabolite of glycine, was elevated in plasma, CSF, and brain tissue. Glycine uptake in brain tissue increased in a dose-dependent manner. Time concentration curves revealed that glycine was most likely transported via the blood-CSF barrier and activated NMDA receptors adjacent to the ventricles. The pharmacokinetic analysis and the brain uptake index for glycine suggested that glycine was transported into brain tissue by passive diffusion. These results provide further insight into the potential therapeutic applications of glycine. PMID- 21647663 TI - Editorial to the special issue foldamers. PMID- 21647664 TI - Studies of H4R antagonists using 3D-QSAR, molecular docking and molecular dynamics. AB - Three-dimensional quantitative structure-activity relationship studies were performed on a series of 88 histamine receptor 4 (H4R) antagonists in an attempt to elucidate the 3D structural features required for activity. Several in silico modeling approaches, including comparative molecular field analysis (CoMFA), comparative similarity indices analysis (CoMSIA), molecular docking, and molecular dynamics (MD), were carried out. The results show that both the ligand based CoMFA model (Q (2) = 0.548, R (ncv) (2) = 0.870, R (pre) (2) = 0.879, SEE = 0.410, SEP = 0.386) and the CoMSIA model (Q (2) = 0.526, R (ncv) (2) =0.866, R (pre) (2) = 0.848, SEE = 0.416, SEP = 0.413) are acceptable, as they show good predictive capabilities. Furthermore, a combined analysis incorporating CoMFA, CoMSIA contour maps and MD results shows that (1) compounds with bulky or hydrophobic substituents at positions 4-6 in ring A (R2 substituent), positively charged or hydrogen-bonding (HB) donor groups in the R1 substituent, and hydrophilic or HB acceptor groups in ring C show enhanced biological activities, and (2) the key amino acids in the binding pocket are TRP67, LEU71, ASP94, TYR95, PHE263 and GLN266. To our best knowledge, this work is the first to report the 3D QSAR modeling of these H4R antagonists. The conclusions of this work may lead to a better understanding of the mechanism of antagonism and aid in the design of new, more potent H4R antagonists. PMID- 21647665 TI - Analysis of the classical, alternative, and mannose binding lectin pathway of the complement system in the pathogenesis of oligoarticular juvenile idiopathic arthritis. AB - The complement system plays a role in the pathogenesis of some autoimmunopathies. This longitudinal study evaluates the contribution of the complement system in the pathogenesis of oligoarticular juvenile idiopathic arthritis (JIA). Serum of the peripheral blood and the synovial fluid were investigated for the activity of the classical (CP), the mannose binding lectin (MBL), and the alternative pathway (AP). A total of 12 samples from peripheral blood (PB) and two samples from synovial fluid (SF) of girls with oligoarticular JIA were investigated in a longitudinal observation from the time point of the diagnosis of JIA. The differences between the complement activity in the PB and in the SF were extremely statistically significant (CP and MBL: P < 0.0001; AP: < 0.0087). The activity of the CP and the MBL pathway was reduced. The AP is the main contributor in the pathogenesis of oligoarticular JIA. Anti-C5 therapy may be an option to avoid the creation of the membrane attack complex. PMID- 21647666 TI - [Long-term consequences of postoperative delirium]. AB - A patient reported anxiety and sleeping problems 9 months after reconstruction of the anterior floor of the mouth following tumor surgery. These symptoms had been initiated by a postoperative delirium with hallucinations, which had not been detected during its occurrence. One session of psychotherapy 9 months later reduced the symptoms. Patients in intensive care units should be asked and informed about delirium symptoms. This might prevent long-term psychological distress. PMID- 21647667 TI - Severe nutritional iron-deficiency anaemia has a negative effect on some bone turnover biomarkers in rats. AB - BACKGROUND: The role of iron (Fe) in bone formation and disease have not received much attention, a fact that is interesting given the known biochemical role that this mineral has upon collagen maturation together with the high prevalence of Fe deficiency anaemia worldwide. AIM: To investigate the changes in bone formation, resorption and mineral content in developing rats with induced nutritional Fe deficiency anaemia. METHODS: Thirty male Wistar rats were divided into two groups, a control group receiving AIN-93G diet with normal-Fe content and an anaemic group receiving AIN-93G diet with low-Fe content for 40 days. Both diets were prepared with an adequate calcium (Ca) and phosphorus (P) content. The most representative serum bone turnover biomarkers and femur and sternum calcium and phosphorus content, together with sternum Fe content were determined in both experimental groups. RESULTS: In anaemic rats, bone matrix formation diminished as revealed by the lower amount of procollagen type I N-terminal propeptide. Bone resorption process increased in Fe deficiency as shown by the increase of serum parathyroid hormone, tartrate-resistant acid phosphatase and levels of degradation products from C-terminal telopeptides of type I collagen released to the serum. In addition, mineralization process was affected by Fe deficiency, because Ca and P content in femur decreased markedly. CONCLUSIONS: Fe-deficiency anaemia had a significant impact upon bone, affecting bone mineralization, decreasing the matrix formation and increasing bone resorption, therefore it is of great interest to assess bone status in situation of Fe-deficiency anaemia. PMID- 21647668 TI - Effect of the consumption of a fruit and vegetable soup with high in vitro carotenoid bioaccessibility on serum carotenoid concentrations and markers of oxidative stress in young men. AB - AIM: To evaluate the effect of the daily intake of a fruit & vegetable soup with high in vitro bioaccessibility of carotenoids on beta-carotene and lycopene serum concentrations. METHODS: Fourteen healthy young men (24 +/- 1 years) received 300 mL/day of a carrot, tomato, and broccoli soup, containing 3.9 mg beta-carotene and 4 mg lycopene, for 4 weeks followed by a 4-week washout period. The serum carotenoid response and oxidative markers were analyzed after 3 and 4 weeks of soup consumption and after a 4-week washout. RESULTS: The in vitro bioaccessibility of beta-carotene and lycopene was 55 and 43%, respectively, in the soup. Serum beta-carotene concentrations were significantly higher than baseline (0.33 +/- 0.05 MUmol/L) after 3 weeks (0.69 +/- 0.06 MUmol/L) and 4 weeks (0.78 +/- 0.10 MUmol/L) of soup consumption (P < 0.001). Serum lycopene was also significantly higher compared with baseline levels (0.26 +/- 0.08-0.56 +/- 0.04 MUmol/L and 0.60 +/- 0.04 MUmol/L, after 3 and 4 weeks, respectively) (P < 0.001). Although the highest concentration of both carotenoids was found after 4 weeks, the levels were not statistically different from the levels at 3 weeks. A 4-week washout significantly decreased serum carotenoid concentrations, although only beta-carotene returned to baseline. Glutathione peroxidase (GPx) increased significantly after soup supplementation compared with baseline, while superoxide dismutase was significantly lower only after 3 weeks. Glutathione reductase, lipid, protein, and DNA oxidative markers remained unchanged. CONCLUSIONS: The soup contributed to increasing the concentration of each carotenoid by more than 100% after 3 and 4 weeks of consumption, the maximum increase being observed after 4 weeks. Oxidative markers did not show any variation except for GPx. Serum lycopene half-life was longer than that of beta-carotene, which may be important for studies evaluating both carotenoids. PMID- 21647669 TI - Influence of gender on DHA synthesis: the response of rat liver to low dietary alpha-linolenic acid evidences higher omega3 ?4-desaturation index in females. AB - PURPOSE: The conversion rate of alpha-linolenic acid (ALA) into docosahexaenoic acid (DHA) is determined by dietary and non-dietary factors. Higher capacity of DHA synthesis has been evidenced in females, indicating that sex factors influence the conversion pathway. To evaluate the extent to which sexual dimorphism of DHA synthesis is subordinated to nutritional handling, we measured the omega3 ?4-desaturation index in male and female rats receiving adequate or inadequate amounts of ALA. The omega3 ?4-desaturation index was drawn from the DHA to docosapentaenoic acid (omega3DPA) ratio in liver phospholipids. METHODS: Male and female rats born to omega3-deficient dams were fed a supplemented diet supplying low, inadequate, intermediate, or adequate ALA (5, 20, 100, or 300 mg ALA/100 g diet, respectively). Control rats from both gender received the adequate diet from fetal life. RESULTS: Compared with control, low ALA feeding induced the omega3 ?4-desaturation index to increase by 38 and 70% in the phosphatidylethanolamine fraction of males and females, respectively, and by 67% in phosphatidylcholine in females only. Supplementations with increased doses of ALA progressively smoothed this gender effect. Moreover, the analysis of our data from a previous study shows that ovariectomy decreased, whereas estradiol treatment increased the omega3 index to values comparable with those of diet matched males and intact females, respectively. CONCLUSION: Females are more prone than males to increase their index of omega3 ?4-desaturation, especially in response to low supplies in ALA. Estradiol supports the omega3 index, suggesting that this hormone plays a role in the effect of gender on DHA synthesis. PMID- 21647670 TI - Evaluation of culture filtrates of Culicinomyces clavisporus: Mycoadulticide for Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi. AB - The Culicinomyces clavisporus is a fungal pathogen of a wide range of mosquito larvae. The C. clavisporus was isolated from the larvae of Culiseta inornata. We have investigated into potential pathogenicity against the adults of Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi. The culture filtrates released from the strain of C. clavisporus 46258 were grown in the EmYPss broth, were filtered and used for the bioassays after a growth of 15 days. The results demonstrated these metabolites with LC(50), LC(90) and LC(99) values of C. quinquefasciatus, 5.62, 8.71 and 12.59, A. aegypti, 3.0, 7.0 and 9.3, and A. stephensi 2.69, 6.0 and 7.24 MUl/cm(2), respectively after exposure for 24 h. These results compared favorably with the commercial adulticide Gokilaht((r))-S 5EC (d,d-trans-cyphenothrin) that showed 100% mortality at the same concentration. This study successfully identified that the metabolites of C. clavisporus can be used as mosquitoes adulticide as safer alternative to modern synthetic chemical insecticide against mosquito vector of diseases. Further purification can lead to biotechnological exploitation. PMID- 21647671 TI - Coccidiosis of domestic rabbit (Oryctolagus cuniculus) in Egypt: light microscopic study. AB - In the present study, the incidence and prevalence of coccidian infection among domestic rabbits in Egypt were investigated. Severe overall prevalence reaching 70% (70/100) was recorded. Eight species of Eimeria were detected. Mixed infection with three different species occurred most frequently. Eimeria intestinalis and Eimeria coecicola were generally the most predominant species. The complete life cycle of E. intestinalis was investigated. This study is the first to report coccidia in domestic rabbits in Egypt. Six species of Eimeria were reported for the first time. PMID- 21647672 TI - Morphobiometrical and molecular study of two populations of Demodex folliculorum from humans. AB - A morphobiometrical and molecular study of two populations of Demodex folliculorum from humans isolated from different habitats, skin and eyelashes follicles, were carried out. Morphological and biometrical studies revealed two closely related populations with any distinctive characteristics. For molecular study, a 436-bp region of the 16S rDNA gene and a 453-bp region of the cytochrome oxidase I (COI) gene from individual mites of each population considered were sequenced. Intraindividual and interindividual sequence variation was studied in both populations. Our data show that 16S rDNA is not a useful marker to discriminate between populations; however, COI gene sequences can help to identify the two populations considered, which are morphologically very close and difficult to separate by classic methods. These results are in agreement with the morphological and biometrical differences detected between D. folliculorum from eyelashes and human skin. This study appeals for the revision of the taxonomic status of the D. folliculorum populations, as well as for the species included within genus Demodex. PMID- 21647673 TI - The antiplasmodial activity of Anogeissus leiocarpus and its effect on oxidative stress and lipid profile in mice infected with Plasmodium bergheii. AB - Methanolic extracts of Anogeissus leiocarpus has been considered locally to have the same antimalarial activities as artemisinin derivatives. This work studied the in vivo antiplasmodial activity of methanolic extracts of A. leiocarpus and its effect on oxidative stress and lipid profile in mice infected with Plasmodium bergheii. Mice used for this study were divided into five groups; four of the groups were infected with P. bergheii. The first group was not infected with the parasite. The second group was infected with parasite but not treated with antimalarial drugs (negative control). The third group was infected and treated with artesunat at 5 mg/kg body weight (positive control), while the fourth and fifth groups were infected and treated with 100 and 200 mg/kg body weight of extract of stem bark of A. leiocarpus, respectively. The rate of parasite clearance was higher in the group treated with 200 mg/kg body weight of extract of A. leiocarpus when compared with the groups treated with artesunat. Malondialdehyde (MDA) level was significantly higher (P < 0.05) in the serum of negative control as compared with other groups which have received treatment. MDA level was moderately higher in the liver homogenates of infected mice treated with artesunat than in other groups. There were significant increases (P < 0.05) in the levels of serum and liver superoxide dismutase of infected mice treated with 200 mg/kg body weight of A. leiocarpus when compared with other groups. Serum low density lipoprotein, total triglyceride, and total cholesterol were moderately higher in the group treated with artesunat than other groups, while high density lipoprotein (HDL) level was higher in the two groups treated with A. leiocarpus as compared with the group treated with artesunat. This study shows that the methanolic extract of A. leiocarpus has high antimalarial activities, high antioxidant property, and capable of boosting HDL level in malaria-infected organisms. PMID- 21647674 TI - Efficacy of fungus mediated silver and gold nanoparticles against Aedes aegypti larvae. AB - Chrysosporium tropicum is a pathogenic fungus. It is known to be an effective mosquito control agent. In the present study, we have synthesized the silver and gold nanoparticles using C. tropicum. These nanoparticles have been characterized through Microscan reader, X-ray diffractometer, transmission electron microscopy, and further confirmed by scanning electron microscopy. The characterization study confirmed the spherical shape and size (2-15 and 20-50 nm) of gold and silver nanoparticles. These silver and gold nanoparticles have been tested as a larvicide against the Aedes aegypti larvae. The larvicidal efficacy was noted when performed against all instars of A. aegypti at six different log concentrations, and significant results could be observed. The gold nanoparticles used as an efficacy enhancer have shown mortality at three times higher concentration than the silver nanoparticles. The larval mortality was observed after different time of exposures. The mortality values were obtained using the probit analysis. The larvae of A. aegypti were found to be highly susceptible for the silver nanoparticles. The second instar larvae have shown 100% mortality against the silver nanoparticles after 1 h, whereas the first, third, and fourth instars have shown efficacy (LC(50) = 3.47, 4, and 2; LC(90) = 12.30, 8.91, and 4; LC(99) = 13.18, 13.18, and 7.58, respectively) after 1 h. The results could suggest that the use of fungus C. tropicum, silver, and gold nanoparticles is a rapid, environmentally safer, and greener approach for mosquito control. This could lead us to a new possibility in vector control strategy. PMID- 21647675 TI - Surface features of the monogenean gill parasites Pseudodactylogyrus anguillae and Pseudodactylogyrus bini from the European eel Anguilla anguilla in Egypt. AB - In the present study, scanning electron microscopy is used to demonstrate the surface features of Pseudodactylogyrus anguillae Yin and Sproston (1948) Gussev, 1965 and Pseudodactylogyrus bini Kikuchi (1929) Gussev, 1965 from the European eel Anguilla anguilla. Specimens of the eel A. anguilla were collected from the River Nile, near Mansoura, Dakahlia Province, Egypt. Morphologically, P. anguillae could be distinguished from P. bini based on the total length, the shape and size of the haptor, and the constriction between the haptor and body proper. The general body tegument and the haptoral tegument of both species are microvillous. Possible functions of the microvilli associated with the tegument covering the anterior adhesive areas are discussed. The region of the body proper just anterior to the haptor of P. anguillae, appears to be highly folded and provided with many projections forming scale-like structures. Many surface ciliary sensilla presumed to be sensory structures are found on the tegument of the anterior adhesive area of P. anguillae. Secretory bodies and mucus-like substances are recorded on the tegument covering the anterior adhesive areas of P. anguillae and the haptor of P. bini. The possible role of these secretions in the attachment of the anterior adhesive areas and the posterior attachment haptor to the host tissues are also discussed. PMID- 21647676 TI - Androgens and musculoskeletal symptoms among breast cancer patients on aromatase inhibitor therapy. AB - Aromatase inhibitors (AIs), the adjuvant hormonal treatment of choice for postmenopausal estrogen receptor-positive breast cancer, are associated with an increased risk of musculoskeletal symptoms. The underlying cause of the symptoms is often attributed to estrogen depletion, yet all women treated with AIs have low estrogen levels and only a subset develop symptoms. Concentrations of circulating androgens may be mediating factors contributing to these side effects. The purpose of this study was to examine changes in androgen concentrations among women initiating AI therapy and to determine if concentrations are associated with musculoskeletal symptoms. Data were analyzed from a cohort study of 74 breast cancer patients for whom AI therapy was planned. Questionnaire data on symptoms were collected and blood was drawn prior to AI therapy (baseline) and then again at 3 and 6 months after baseline. Blood was assayed for testosterone, androstenedione, dehydroepiandrosterone-sulfate (DHEAS), and sex hormone-binding globulin (SHBG). Free testosterone index (FTI) values were calculated using testosterone and SHBG measurements. The results showed that concentrations of all of the androgens increased over the study period, with statistically significant differences from baseline concentrations observed for the FTI at 3 and 6 months and for DHEAS at 6 months. Additionally, breast cancer patients with new onset or worsening of pain over the study period had a significantly smaller change in mean DHEAS concentration from baseline to 3 months (P = 0.04) and a marginally significant smaller change in mean DHEAS concentration from baseline to 6 months (P = 0.1) compared to those who reported no pain at all time points or no worsening of pain across the study period. Changes in testosterone, androstenedione, and the FTI were not associated with the onset or worsening of pain during the study period. Findings from this study suggest that higher DHEAS concentrations are associated with less AI-associated pain and should be further investigated. PMID- 21647677 TI - Pilot and feasibility study: prospective proteomic profiling of mammary epithelial cells from high-risk women provides evidence of activation of pro survival pathways. AB - Normal mammary gland homeostasis requires the coordinated regulation of protein signaling networks. However, we have little prospective information on whether activation of protein signaling occurs in premalignant mammary epithelial cells, as represented by cells with cytological atypia from women who are at high risk for breast cancer. This information is critical for understanding the role of deregulated signaling pathways in the initiation of breast cancer and for developing targeted prevention and/or treatment strategies for breast cancer in the future. In this pilot and feasibility study, we examined the expression of 52 phosphorylated, total, and cleaved proteins in 31 microdissected Random Periareolar Fine Needle Aspiration (RPFNA) samples by high-throughput Reverse Phase Protein Microarray. Unsupervised hierarchical clustering analysis indicated the presence of four clusters of proteins that represent the following signaling pathways: (1) receptor tyrosine kinase/Akt/mammalian target of rapamycin (RTK/Akt/mTOR), (2) RTK/Akt/extracellular signal-regulated kinase (RTK/Akt/ERK), (3) mitochondrial apoptosis, and (4) indeterminate. Clusters 1 through 3 comprised moderately to highly expressed proteins, while Cluster 4 comprised proteins that are lowly expressed in a majority of RPFNA samples. Our exploratory study showed that the interlinked components of mitochondrial apoptosis pathway are highly expressed in all mammary epithelial cells obtained from high-risk women. In particular, the expression levels of anti-apoptotic Bcl-xL and pro apoptotic Bad are positively correlated in both non-atypical and atypical samples (unadjusted P < 0.0001), suggesting a delicate balance between the pro-apoptotic and anti-apoptotic regulation of cell proliferation during the early steps of mammary carcinogenesis. Our feasibility study suggests that the activation of key proteins along the RTK/Akt pathway may tip this balance to cell survival. Taken together, our results demonstrate the feasibility of mapping proteomic signaling networks in limited RPFNA samples obtained from high-risk women and the promise of developing rational drug targets or preventative strategies for breast cancer in future proteomic studies with a larger cohort of high-risk women. PMID- 21647678 TI - Expression of claudin 10 protein in hepatocellular carcinoma: impact on survival. AB - PURPOSE: To investigate the expression of claudin 10 in hepatocellular carcinoma (HCC) and the impact on angiogenesis and the postoperative survival of HCC patients. METHODS: The expression of claudin 10 protein was analyzed on samples from 99 HCC patients undergoing hepatectomy with immunohistochemistry and 31 fresh specimens with Western blotting. We examined the relationship between claudin 10 expression with clinicopathological factors, microvessel density (MVD), and postoperative survival. RESULTS: Western blotting and immunohistochemical staining showed that claudin 10 protein was highly expressed in HCC, compared with paraneoplastic liver tissue and normal liver tissue (P < 0.01). Claudin 10 protein expression levels were significantly higher in HCC specimens with microscopic venous invasion (P < 0.01). MVD in HCC increased with enhanced claudin 10 expression (P < 0.01). Kaplan-Meier survival analysis revealed that HCC patients with high claudin 10 expression had significantly shorter overall survival (P = 0.01). Multivariate analysis showed that claudin 10 expression was an independent prognostic indicator for postoperative overall survival of HCC patients (P < 0.01). CONCLUSIONS: Claudin 10 protein is highly expressed in HCC tissue and is closely related to angiogenesis. Claudin 10 protein could be a useful marker to predict poor prognosis of HCC patients after hepatectomy. PMID- 21647679 TI - Folding dynamics of phenylalanine hydroxylase depends on the enzyme's metallation state: the native metal, iron, protects against aggregate intermediates. AB - Phenylalanine hydroxylase (PAH), a non-heme iron enzyme, is responsible for the phenylalanine conversion to tyrosine. Its malfunction causes phenylketonuria (PKU). To better understand how protein structure and folding profiles are affected by the metal cofactor, we investigated the chemical (un)folding of apo- and holo-PAH from Chromobacterium violaceum (cPAH) using circular dichroism (CD) and analytical ultracentrifugation (AUC). Holo-cPAH shows a two-state unfolding transition. In contrast, the unfolding profile for apo-cPAH reveals a three-state (un)folding pathway and accumulation of an intermediate (apo-cPAH(I)). This intermediate is also observed in refolding experiments. Fluorescence studies are consistent with the CD findings. The intermediate apo-cPAH(I) and unfolded state(s) of apo- and holo-cPAH(U) have been characterized by analytical ultracentrifugation (AUC). At 2.4 and 2.8 M GuHCl, 90% of the signal for apo-cPAH has a weight average sedimentation coefficient in water at 20 degrees C (s20,w) of about 48 S, representing multiple aggregate species made of multiple monomers of cPAH. Aggregate formation for apo-cPAH is also confirmed by dynamic light scattering and electron microscopy giving a hydrodynamic radius (R(H)) of 41 nm for apo-cPAH(I) versus 3.5 nm for the native protein. PMID- 21647680 TI - Real-time marker-free patient registration for electromagnetic navigated bronchoscopy: a phantom study. AB - PURPOSE: To perform bronchoscopy safely and smoothly, it is very important to develop a bronchoscopic guidance system. Transbronchial lung biopsy (TBLB) with a bronchoscopic guidance system especially should permit safe image-guided procedures. Recently, electromagnetic tracking (EMT) is utilized to track the tip of the bronchoscope camera in real time. For most tracking methods using position sensors, registration between tracking data and previously acquired reference image data, such as CT image, is performed using natural landmarks of the patient or fiducial markers attached to the patient, whose positions need to be measured manually by the physician before the actual bronchoscopy. Therefore, this paper proposes a marker-free CT-to-patient registration method utilizing bronchoscope's position and orientation obtained by the EMT. METHODS: We developed a guidance system that is able to track the tip of the bronchoscope camera in real time. In the case of a guidance system that uses position sensors, natural landmarks of the patient or fiducial markers attached to the patient are needed to obtain the correspondence between EMT outputs and previously acquired reference image data, such as CT image. This paper proposes a registration method without landmarks or fiducials by estimating the transformation matrix between the patient and the CT image taken prior to the bronchoscopic examination. This estimation is performed by computing correspondences between the outputs of the EMT sensor and airways extracted from the CT image. As ambiguities between EMT measurements and their corresponding airway branches may arise at airway bifurcations, we introduce a stable airway branch selection mechanism for improving the robustness of the estimation of the transformation matrix. To evaluate the performance of the proposed method, we applied the method to a rubber bronchial phantom and added virtual breathing motion to the sensor output. RESULTS: Experimental results show that the accuracy of our proposed method is within 2.0-3.0 mm (without breathing motion) and 2.5-3.5 mm (with breathing motion). The proposed method could also track a bronchoscope camera in real time. CONCLUSIONS: We developed a method for CT-to-patient registration using a position sensor without fiducial markers and natural landmarks. Endoscopic guided biopsy of lung lesions is feasible using a marker-free CT-to-patient registration method. PMID- 21647681 TI - Optimization of fibrolytic enzyme production by Aspergillus japonicus C03 with potential application in ruminant feed and their effects on tropical forages hydrolysis. AB - Fibrolytic enzyme production by Aspergillus japonicus C03 was optimized in a medium containing agro-industrial wastes, supplemented with peptone and yeast extract. A 2(3) full factorial composite and response surface methodology were used to design the experiments and analysis of results. Tropical forages were hydrolyzed by A. japonicus C03 enzymatic extract in different levels, and they were also tested as enzymatic substrate. Optimal production to xylanase was obtained with soybean bran added to crushed corncob (1:3), 0.01% peptone, and 0.2% yeast extract, initial pH 5.0, at 30 degrees C under static conditions for 5 days of incubation. Optimal endoglucanase production was obtained with wheat bran added to sugarcane bagasse (3:1), 0.01% peptone, and 0.2% yeast extract, initial pH 4.0, at 30 degrees C, for 6 days, under static conditions. Addition of nitrogen sources as ammonium salts either inhibited or did not influence xylanase production. This enzymatic extract had a good result on tropical forage hydrolyzes and showed better performance in the Brachiaria genera, due to their low cell wall lignin quantity. These results represent a step forward toward the use of low-cost agricultural residues for the production of valuable enzymes with potential application in animal feed, using fermentation conditions. PMID- 21647682 TI - Alzheimer disease: are we intervening too late? Pro. AB - The affirmative position is argued in response to the question of whether intervention in the disease course of Alzheimer disease (AD) occurs too late. AD is not a singular, homogeneous disease, but rather a final common pathway or end point that can be arrived at through multiple routes. As part of the affirmative argument, there is a delineation of two long-term trajectories leading to AD: (1) normal elderly progression to AD, and (2) depressed elderly progression to AD. In documenting normal elderly devolution into AD, two "normal" elderly pre-AD or prodromal stages are discussed: age-associated memory impairment (AAMI) and mild cognitive impairment (MCI). Data are provided evidencing significantly high conversion rates from these pre-AD stages to actual AD. Using the same paradigmatic approach that is used in documenting normal elderly decline into AAMI and MCI with eventual conversion to AD; there is explication of depressed elderly conversion to AD. The long-term, multiphasic disease progression of major depression without dementia to depressive dementia to final conversion to AD is brought into focus as another example of why intervention must occur prior to actual conversion to AD. Depression is defined as a cognitive syndrome and risk factor for AD requiring aggressive targeted intervention. AD does not just come suddenly out of nowhere. First intervention must occur during the pre-AD phases in an attempt to prevent, delay, and interrupt long-term neurodegenerative processes involved in both normal elderly and depressed elderly conversion to AD. A primary strategy proposed is to delay onset of AD. Population statistics indicate that if AD is delayed by a modest 1 year, there would be 9.5 million fewer cases by 2050, resulting in significant reduction in burden of disease. Data show early intervention with cognitive stimulation (mental exercise), physical exercise, aggressive treatment of AD risk factors and excess disability, psychotherapy, and other nonpharmacological interventions in combination with each other and/or with medications can result in delay of onset of AD. First intervention at time of diagnosis of AD is too late, when by definition, final conversion to AD has already occurred. When we have knowledge to successfully intervene earlier, why would we not want to do so. PMID- 21647683 TI - Normal sensory and absent cognitive electrophysiological responses in functional visual loss following chemical eye burn. AB - OBJECTIVE: To present a unique case of a 34-year-old patient with unilateral functional visual loss after chemical burn with normal visual evoked potentials (VEPs) and absent cognitive response (P300 wave). METHODS: Visual functions, complete ophthalmic and neurologic examinations including computed tomography of the brain, electrophysiological testing of the visual pathway up to the cognitive brain cortex were evaluated. Data were collected prospectively during 1-year follow-up and compared with data from published case series and a literature review. RESULTS: No abnormalities were found that could account for such a rapid monocular loss of vision with exception of absence of the P300 wave in the affected eye during cognitive tasks. Vision slowly improved during 1 year without any treatment. CONCLUSIONS: Functional vision loss is a diagnosis of exclusion. In the event of reduced vision in the context of a normal ocular health examination, all other pathology must be ruled out before the diagnosis of functional visual loss is established. Complex visual electrophysiological testing is the preferred tool for objective examination of such disorders. PMID- 21647684 TI - Reduction of enzyme dosage by oxygen delignification and mechanical refining for enzymatic hydrolysis of green liquor-pretreated hardwood. AB - In this study, a strategy to reduce enzyme dosage is evaluated by applying two post-treatments, oxygen delignification and mechanical refining. The sugar conversion for GL12 substrates was increased from 51.5% to 77.9% with post treatments at the enzyme dosage of 10 FPU. When the amount of enzyme was reduced to 5 FPU with post-treatments, the conversion of 71.8% was obtained, which was significant higher than the conversion without any post-treatment using 10 FPU (51.5%). This clearly demonstrates the benefit of post-treatments that allows more than 50% of enzyme reduction at the same level of enzymatic conversion. Enzyme-accessible surface area and pore volume were evaluated by Simons' staining and DSC thermoporometry methods, and strong correlations were found with the sugar conversion. PMID- 21647685 TI - An improved procedure for the purification of catalytically active alkane hydroxylase from Pseudomonas putida GPo1. AB - Bacterial alkane hydroxylases are of high interest for bioremediation applications as they allow some bacteria to grow in oil-contaminated environments. Furthermore, they have tremendous biotechnological potential as they catalyse the stereo- and regio-specific hydroxylation of chemically inert alkanes, which can then be used in the synthesis of pharmaceuticals and other high-cost chemicals. Despite their potential, progress on the detailed characterization of these systems has so far been slow mainly due to the lack of a robust procedure to purify its membrane protein component, monooxygenase AlkB, in a stable and active form. This study reports a new method for isolating milligramme amounts of recombinant Pseudomonas putida GPo1 AlkB in a folded, catalytically active form to purity levels above 90%. AlkB solubilised and purified in the detergent lauryldimethylamine oxide was demonstrated to be active in catalysing the epoxidation reaction of 1-octene with an estimated K (m) value of 0.2 mM. PMID- 21647686 TI - Analysis of the amino acids of soy globulins by AOT reverse micelles and aqueous buffer. AB - The 7S and 11S globulins from soybean proteins using reverse micelle and aqueous buffer extraction methods were characterized by using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and scanning electron microscope (SEM), and their amino acid compositions were also evaluated. SDS-PAGE did not show electrophoretic differences between 7S and 11S globulin subunits with two extraction methods. SEM analysis showed that the AOT reverse micelle processing of 7S and 11S globulins induced a reduction of droplet size. Some individual amino acid contents of 7S and 11S globulins using two extraction methods were different, some were similar. In all the samples, the glutamic acid, aspartic acid, and leucine were the dominant amino acids while the cystine and methionine were the first-limiting amino acids. The proportion of essential amino acids to the total amino acids (E/T) of the 7S globulin from aqueous buffer and reverse micelles was similar. While significant differences were obtained in the proportion of E/T of the 11S globulin. PMID- 21647687 TI - Investigation of the effective action distance between hematopoietic stem/progenitor cells and human adipose-derived stem cells during their in vitro co-culture. AB - The in vitro suitable action distance between umbilical cord blood-derived hematopoietic stem/progenitor cells and its feeder cell, human adipose-derived stem cells, during their co-culture, was investigated through a novel transwell co-culture protocol, in which the distance between the two culture chambers where each cell type is growing can be adjusted from 10 to 450 MUm. The total cell number was determined with a hemacytometer, and the cell morphology was observed under an inverted microscope each day. After 7 days of co-culture, the fold expansion, surface antigen expression of CD34(+) and CFU-GM assay of the hematopoietic mononuclear cells (MNCs) were analyzed. The results showed that there was an optimal communication distance at around 350 MUm between both types of stem cells during their in vitro co-culture. By using this distance, the UCB MNCs and CD34(+) cells were expanded by 15.1 +/- 0.2 and 5.0 +/- 0.1-fold, respectively. It can therefore be concluded that the optimal action distance between stem cells and their supportive cells, when cultured together for 7 days, is of around 350 MUm. PMID- 21647688 TI - Co-cultured production of lignin-modifying enzymes with white-rot fungi. AB - Co-cultivation was a potential strategy in lignocellulolytic biodegradation with producing high activity enzymes due to their synergistic action. The objective of this study was to investigate the rarely understood effects of co-culturing of two white-rot fungi on lignin-modifying enzymes (LMEs) production. Six species, Bjerkandera adusta, Phlebia radiata, Pleurotus ostreatus, Dichomitus squalens, Hypoxylon fragiforme and Pleurotus eryngii, were cultured in pairs to study the production of LMEs. The paired hyphal interaction observed showed that P. eryngii is not suitable for co-growth. The use of agar plates containing dye RBBR showed elevated decolourisation at the confrontation zone between mycelia. Laccase was significantly stimulated only in the co-culture of P. radiata with D. squalens under submerged cultivation; the highest value was measured after 4 days of incubation (120 U mg(-1)). The improved productions of MnP and LiP were simultaneously observed at the co-culture of P. ostreatus and P. radiata (MnP = 800 nkat L(-1) after 4 days of incubation; LiP = 60 nkat L(-1) after 7 days of incubation), though it was not a good producer of laccase. P. ostreatus appeared to possess specific potential to be used in co-cultured production of LMEs. The phenotype of LMEs production was not only dependent on the species used but also regulated by different nutritions available in the culture medium. The present data will provide evidence for illustrating the regulatory roles of C/N on LMEs production under the co-cultures' circumstances. PMID- 21647689 TI - The health of irregular and illegal immigrants: analysis of day-hospital admissions in a department of migration medicine. AB - It is difficult to trace full details of the path which irregular or illegal immigrants follow when seeking assistance in the network of the various hospital departments and health structures. The aim of this work was to analyze the health needs of immigrant people by reviewing the types of treatment given to them in the day-hospital of our Department of Migration Medicine. Our study analyzed day hospital admissions between 2003 and 2009. The patient charts used for managing day-hospital activity were adopted in 2002 in conformity with the "OSI project". From these it is possible to draw up a scale picture of the distribution of each pathology in the immigrant population. The sample population consisted of 1,758 subjects, representing 7.4% of potential users. More than half came from Africa, followed by Asia, and then Europe. Gastroenterological diseases ranked first, with dyspeptic syndromes most frequently diagnosed. Infections and parasitic diseases ranked second, and the most frequent diagnoses were sexually transmitted diseases. Third were diseases of the genitourinary system. Metabolic disorders ranked fourth, among them, more than half of the cases were of diabetes mellitus, in patients from south-east Asia. Diseases of the circulatory system were sixth, with hypertension the most frequent pathology. Our data confirm a marked persistence of the phenomenon known as the "healthy immigrant effect" in these types of patients, as well as the prominent role played by "social determinants" in conditioning the health of immigrants, particularly in the case of some infectious diseases. PMID- 21647690 TI - Early invasive strategy and outcomes of non-ST-elevation acute coronary syndrome patients: is time really the major determinant? AB - In non-ST-elevation acute coronary syndromes (ACS), an early invasive strategy is recommended for middle/high-risk patients; however, the optimal timing for coronary angiography is still debated. The aim of this study was to evaluate the prognostic implications of the time of angiography in ACS patients treated in accord with an early invasive strategy. We analyzed the relationship between the time of angiography and outcomes at follow-up in 517 ACS patients, of whom 482 were revascularized with percutaneous coronary intervention (PCI) (86.9%) or coronary artery by-pass graft (13.1%). We also evaluated the influence of clinical, biohumoral and angiographic variables on the patients' outcomes at follow-up. Among patients submitted to angiography at different time intervals from both hospital admission and symptom onset, significant differences neither in mortality nor in cardiac ischemic events at follow-up were observed. At univariate analysis, complete versus partial revascularization, longer hospital stay, higher TIMI risk score, diabetes mellitus, higher discharge creatinine and admission anemia were associated with mortality and cardiac ischemic events at follow-up; a lower left ventricular ejection fraction was associated with mortality; higher peak troponin I and previous PCI were associated with cardiac ischemic events at follow-up. At multivariate analysis longer hospital stay, higher discharge creatinine levels, and previous PCI were independent predictors of cardiac ischemic events at follow-up. Our evaluation in ACS patients treated with an early invasive strategy does not support the concept that angiography should be performed as soon as possible after symptom onset or hospital admission. Rather, an unfavorable long-term outcome is influenced principally by the clinical complexity of patients. PMID- 21647691 TI - Pilot trial of interleukin-2 and zoledronic acid to augment gammadelta T cells as treatment for patients with refractory renal cell carcinoma. AB - Prior to the advent of VEGF-targeted therapies, renal cell carcinoma (RCC) was among the few solid tumors shown to respond to cytokine-based therapies such as interleukin-2 (IL-2) and interferon alpha. Previous work has shown that aminobisphosphonates, including zoledronic acid (ZA), are capable of activating human Vgamma9 Vdelta2 T cells in vitro, and these cells can be further expanded with IL-2. Moreover, these Vgamma9 Vdelta2 T cells have cytolytic activity in vitro to multiple human tumor cell lines. In the current report, we have conducted a pilot trial in patients with metastatic RCC, evaluating different doses of ZA in combination with low-dose IL-2 to determine whether combining these agents can promote in vivo proliferation of Vgamma9 Vdelta2 T cells and elicit an antitumor response. In 12 patients evaluated, no objective clinical responses were observed by RECIST criteria; however, two patients experienced prolonged stable disease. A modest increase in Vgamma9 Vdelta2 T-cell frequency could be detected by Day 8 of therapy in four of the nine patients who received at least one cycle of therapy, but not to the magnitude anticipated from preclinical models. Repeated administration of IL-2 and ZA resulted in both a diminished in vivo percentage of Vgamma9 Vdelta2 T cells as well as impaired expansion in vitro after the first cycle of therapy. These results suggest that repeated administration of IL-2 and ZA, at the doses and schedules used in this trial, may actually inhibit the proliferative capacity of Vgamma9 Vdelta2 T cell in patients with metastatic RCC. PMID- 21647692 TI - Optical coherence tomography in dermatology: technical and clinical aspects. AB - Optical coherence tomography (OCT), a fairly new non-invasive optical real-time imaging modality, is an emergent in vivo technique, based on the interference (Michelson interferometry) of infrared radiation and living tissues, that allows high-resolution, 2- or 3-dimensional, cross-sectional visualisation of microstructural morphology of tissues. OCT provides depth-resolved images of tissues with resolution up to a few micrometers and depth up to several millimetres depending on tissue type. The investigations using OCT to assess skin structure in clinical settings started in the past decade and consequently proved that this imaging method is useful in visualizing subsurface structures of normal skin, including the epidermis, dermoepidermal junction, dermis, hair follicles, blood vessels and sweat ducts. An increasing number of papers brought evidence of the utility and the precision of OCT technology, in its different technical variants, in diagnosing and monitoring skin disorders, including malignancies and inflammatory conditions, respectively. The present comprehensive review describes and illustrates technical aspects and clinical applications of OCT methods in dermatology. PMID- 21647693 TI - Combined biodegradation and ozonation for removal of tannins and dyes for the reduction of pollution loads. AB - PURPOSE: Tannins and dyes pose major threat to the environment by generating huge pollution problem. Biodegradation of wattle extract, chrome tannin and dye compounds using suitable fungal culture namely Aspergillus niger, Penicillium sp. were carried out. In addition to these, ozone treatment was carried out to get higher degradation rate. RESULTS: The results were monitored by carrying out chemical oxygen demand (COD), total organic carbon (TOC), and UV-Vis analysis. The results showed that wattle extract (vegetable tannin) gave better biodegradation rate than dye and chromium compounds. Biodegradation plus ozone showed degradation rates of 92-95%, 94-95%, and 85-87% for the wattle extract, dyes, chromium compounds, respectively. UV-Vis showed that there were no peaks observed for biodegraded samples indicating better degradation rates as compared to the control samples. FT-IR spectra analysis suggested that the formation of flavanoid derivatives, chromic oxide and NH(2) compounds during degradation of wattle extract, chromium and dye compounds, respectively, at the peaks of 1,601 1,629 cm(-1), 1,647 cm(-1), and 1,610-1,680 cm(-1). CONCLUSION: The present investigation shows that combination of biodegradation with ozone is the effective method for the removal of dyes and tannins. The biodegradation of the said compounds in combination with ozonation showed better rate of degradation than by chemical methods. The combination of biodegradation with ozone helps to reduce pollution problems in terms of COD, TOC, total dissolved solids and total suspended solids. PMID- 21647694 TI - Aliphatic and polycyclic aromatic hydrocarbons in the surface sediments from the Eastern Aegean: assessment and source recognition of petroleum hydrocarbons. AB - MATERIALS AND METHODS: Aliphatic and polycyclic aromatic hydrocarbons (PAHs) were determined in surficial sediments from the Aegean Sea in the Eastern Mediterranean in 2008. RESULTS: Total aliphatic hydrocarbons (n-C12 to n-C35) ranged from 330 to 2,660 ng g(-1) dry weight (dwt), while aromatics (19 PAHs) varied between 73.5 and 2,170 ng g(-1) dwt. Total concentrations of both aliphatic hydrocarbons and PAHs ranged from a relatively low to a moderate PAHs pollution compared to other urbanized coastal areas worldwide. PAH consisted mainly of pyrolytic four- to five-ring compounds. Both pyrolytic and petrogenic PAHs are present in most samples, although petroleum-derived PAH are dominant at Izmir Inner Bay (IIB) and Dardanelles Strait, and pyrolytic sources are prevalent in other sampling sites. A high contribution of perylene, a diagenetic originated PAH, to the total penta PAHs was found greater than 70% in Meric River Estuary, Dikili Bay, Candarli Bay, and Gokova Bay sites. CONCLUSION: The spatial distributions of aliphatic hydrocarbons and PAHs indicated that urban runoff and transport from the continental shelf is the major input pathway of anthropogenic and biogenic hydrocarbons from terrestrial sources in the near-shore area. PAH levels at all sites were below the effects range-low (ERL) and effects range median (ERM) values except fluorene. The average and maximum fluorene concentrations exceeded ERL, but below ERM, in the IIB. Meanwhile, the concentration levels of naphthalene, fluorene, phenanthrene, fluoranthene, pyrene, benzo[a]anthracene, and chrysene were higher than threshold effect level values at the same site, but all these compounds were significantly lower than the probable effect level values. The results indicated that the sediments should have potential biological impact. PMID- 21647695 TI - Functionally redundant peg sensilla on the scorpion pecten. AB - All scorpions have two mid-ventral organs called pectines. Each pecten has thousands of pore-tipped sensilla sensitive to a variety of volatile organic and water-based stimulants. However, it was previously unknown whether individual sensilla were functionally identical or different. The information enhancement hypothesis predicts that all sensilla have similar chemosensitivities such that each is a unit of a parallel processing system. The information segmentation hypothesis states that sensilla differ in their chemosensitivities, a functional arrangement akin to the glomeruli-specific chemical detection system in the moth or human olfactory sense. In this study, we tested these hypotheses by extracellularly tip-recording sensillar responses to three aqueous tastants: 0.01 M KCl, 0.1 M citric acid, and 40% ethanol by volume. We isolated stimulation to one sensillum at a time and compared the chemoresponses. Sensilla appeared to respond similarly to the same stimulant (i.e., sensillar tip-recordings revealed activity of the same cell types), although sometimes a few sensilla responded with higher spike rates than the others. We conclude that our data primarily support the information enhancement hypothesis but for future tests of sensillar function we suggest a new hybrid model, which proposes that a few specialized sensilla exist among a mostly uniform field of identical sensilla. PMID- 21647696 TI - PCR detection of novel non-ribosomal peptide synthetase genes in lipopeptide producing Pseudomonas. AB - Bacterial lipopeptides (LPs) are a diverse group of secondary metabolites synthesized through one or more non-ribosomal peptide synthetases (NRPSs). In certain genera, such as Pseudomonas and Bacillus, these enzyme systems are often involved in synthesizing biosurfactants or antimicrobial compounds. Several different types of LPs have been reported for non-pathogenic plant-associated Pseudomonas. Focusing on this group of bacteria, we devised and validated a PCR method to detect novel LP-synthesizing NRPS genes by targeting their lipoinitiation and tandem thioesterase domains, thus avoiding amplification of genes for non-LP metabolites, such as the pyoverdine siderophores present in all fluorescent Pseudomonas. This approach enabled detection of as yet unknown NRPS genes in strains producing viscosin, viscosinamide, WLIP, or lokisin. Furthermore, it proved valuable to identify novel candidate LP producers among Pseudomonas rhizosphere isolates. By phylogenetic analysis of these amplicons, several of the corresponding NRPS genes can be tentatively assigned to the viscosin, amphisin, or entolysin biosynthetic groups, while some others may represent novel NRPS systems. PMID- 21647699 TI - Short-term oxygen administration restores blunted baroreflex sensitivity in patients with type 1 diabetes. AB - AIMS/HYPOTHESIS: We hypothesised that the blunted baroreflex sensitivity (BRS) typical of type 1 diabetes is caused by a higher degree of tissue hypoxia in diabetes, and tested whether oxygen increased BRS and ventilation less, equally or more than in healthy control participants (the latter suggesting higher tissue hypoxia). In addition, we also considered the possible interference between oxygen and breathing pattern. METHODS: In 96 participants with type 1 diabetes and 40 age-matched healthy controls, we measured BRS (average of six different standard methods), oxygen saturation, end-tidal carbon dioxide and ventilation changes during spontaneous and controlled breathing at 15 and six breaths/min, in normoxia and during 5 l/min oxygen administration. RESULTS: BRS was blunted and blood pressure higher in diabetic participants during spontaneous breathing (p < 0.05). BRS increased with oxygen during spontaneous breathing in diabetic (p < 0.001) but not in control participants, and with oxygen the difference in BRS was no longer significant. Slow breathing in normoxia restored BRS to a similar extent to giving oxygen. Oxygen increased systolic and diastolic blood pressure, RR interval, heart rate variability, minute ventilation and tidal volume to a greater extent in diabetic patients than in controls, and decreased carbon dioxide similarly to controls. CONCLUSIONS/INTERPRETATION: The increased response to hyperoxia suggests a pre-existing condition of tissue hypoxia that functionally restrains parasympathetic activity in patients with type 1 diabetes. Autonomic abnormalities can be partially and temporarily reversed by functional manoeuvres such as slow breathing or oxygen administration through enhancement of parasympathetic activity and/or correction of tissue hypoxia. PMID- 21647700 TI - Genome-wide association and meta-analysis in populations from Starr County, Texas, and Mexico City identify type 2 diabetes susceptibility loci and enrichment for expression quantitative trait loci in top signals. AB - AIMS/HYPOTHESIS: We conducted genome-wide association studies (GWASs) and expression quantitative trait loci (eQTL) analyses to identify and characterise risk loci for type 2 diabetes in Mexican-Americans from Starr County, TX, USA. METHOD: Using 1.8 million directly interrogated and imputed genotypes in 837 unrelated type 2 diabetes cases and 436 normoglycaemic controls, we conducted Armitage trend tests. To improve power in this population with high disease rates, we also performed ordinal regression including an intermediate class with impaired fasting glucose and/or glucose tolerance. These analyses were followed by meta-analysis with a study of 967 type 2 diabetes cases and 343 normoglycaemic controls from Mexico City, Mexico. RESULT: The top signals (unadjusted p value <1 * 10(-5)) included 49 single nucleotide polymorphisms (SNPs) in eight gene regions (PER3, PARD3B, EPHA4, TOMM7, PTPRD, HNT [also known as RREB1], LOC729993 and IL34) and six intergenic regions. Among these was a missense polymorphism (rs10462020; Gly639Val) in the clock gene PER3, a system recently implicated in diabetes. We also report a second signal (minimum p value 1.52 * 10(-6)) within PTPRD, independent of the previously implicated SNP, in a population of Han Chinese. Top meta-analysis signals included known regions HNF1A and KCNQ1. Annotation of top association signals in both studies revealed a marked excess of trans-acting eQTL in both adipose and muscle tissues. CONCLUSIONS/INTERPRETATION: In the largest study of type 2 diabetes in Mexican populations to date, we identified modest associations of novel and previously reported SNPs. In addition, in our top signals we report significant excess of SNPs that predict transcript levels in muscle and adipose tissues. PMID- 21647701 TI - Review of the European Congress of Radiology musculoskeletal scientific program. PMID- 21647702 TI - Report on the special scientific session held at the ISS Congress, Athens, Greece, 2010. PMID- 21647703 TI - Effect of pan retinal photocoagulation on the serum levels of vascular endothelial growth factor in diabetic patients. AB - This study tests the hypothesis that subjects with proliferative diabetic retinopathy (PDR) have a detectable rise in levels of serum vascular endothelial growth factor (VEGF), which is an important regulator of angiogenesis. Our investigation aims to evaluate plasma VEGF changes after pan-retinal photocoagulation (PRP) in diabetic patients. Twenty-nine type two diabetic patients (17 male, 12 female: mean age 53.13+/-12.22 years) with PDR secondary to diabetes were studied. Blood samples were obtained before and at 2 months after the last PRP session. Serum VEGF levels were measured by ELISA. After PRP, the mean serum VEGF decreased, but this reduction was not remarkable (88.68+/-71.09 vs. 77.01+/-60.33 ng/ml) (P=0.18). There was a statistically significant difference in serum VEGF changes between patients who had regressed PDR with patients who had progressed PDR (-25.98+/-47.37 vs. 56.44+/-31.7 ng/ml) (P=0.003). Our results showed a significant reduction in levels of serum VEGF in the patients who had successful laser treatment. Our findings suggest that serum VEGF levels could be used for monitoring diabetic retinopathy outcome. PMID- 21647704 TI - clinical risk factors for osteoporosis in Ireland and the UK: a comparison of FRAX and QFractureScores. AB - Recently two algorithms have become available to estimate the 10-year probability of fracture in patients suspected to have osteoporosis on the basis of clinical risk factors: the FRAX algorithm and QFractureScores algorithm (QFracture). The aim of this study was to compare the performance of these algorithms in a study of fracture patients and controls recruited from six centers in the United Kingdom and Ireland. A total of 246 postmenopausal women aged 50-85 years who had recently suffered a low-trauma fracture were enrolled and their characteristics were compared with 338 female controls who had never suffered a fracture. Femoral bone mineral density was measured by dual-energy X-ray absorptiometry, and fracture risk was calculated using the FRAX and QFracture algorithms. The FRAX algorithm yielded higher scores for fracture risk than the QFracture algorithm. Accordingly, the risk of major fracture in the overall study group was 9.5% for QFracture compared with 15.2% for FRAX. For hip fracture risk the values were 2.9% and 4.7%, respectively. The correlation between FRAX and QFracture was R = 0.803 for major fracture and R = 0.857 for hip fracture (P <= 0.0001). Both algorithms yielded high specificity but poor sensitivity for prediction of osteoporosis. We conclude that the FRAX and QFracture algorithms yield similar results in the estimation of fracture risk. Both of these tools could be of value in primary care to identify patients in the community at risk of osteoporosis and fragility fractures for further investigation and therapeutic intervention. PMID- 21647705 TI - Adsorption and degradation of zearalenone by bacillus strains. AB - Two Bacillus strains; Bacillus subtilis 168 and Bacillus natto CICC 24640 separately adsorbed and degraded zearalenone in liquid media, in vitro. Viable, autoclaved (121 degrees C, 20 min) and acid-treated cells of both strains separately bound more than 55% of zearalenone (ZEN, 20 MUg/L) after 30 min and 1 h incubation at 37 degrees C under aerobic conditions, and the amount of ZEN adsorbed was dependent on initial cell volume. In addition, ZEN was degraded by the culture extract of both strains. Degradation by B. subtilis 168 and B. natto CICC 24640 culture extract after 24-h aerobic incubation at 30 degrees C was 81% and 100%, respectively. B. natto CICC 24640 culture extract comprehensively degraded ZEN and, for both strains, no oestrogenic ZEN analogues were present. ZEN degradation was accompanied by carbondioxide emission indicating a decarboxylation reaction. ZEN degradation by the salient B. natto CICC 24640 culture extract varied with initial ZEN concentration, incubation time, temperature and pH. Degradation was enhanced by Mn(2+), Zn(2+), Ca(2+) and Mg(2+) but impeded by Hg(2+), Cu(2+), Pb(2+), ethylenediaminetetraacetic acid and 1,10 phenanthroline. The degradation reaction is associated with a metalloproteinase of molar mass in the range 31-43 kDa. Overall, the two generally recognised as safe Bacillus strains can, potentially, be utilised for detoxification of zearalenone in food. PMID- 21647706 TI - Blockade of P2 nucleotide receptors after spinal cord injury reduced the gliotic response and spared tissue. AB - Spinal cord injury (SCI) triggers a sequel of events commonly associated with cell death and dysfunction of glias and neurons surrounding the lesion. Although astrogliosis and glial scar formation have been involved in both damage and repair processes after SCI, their role remains controversial. Our goal was to investigate the effects of the P2 receptors antagonists, PPADS and suramin, in the establishment of the reactive gliosis and the formation of the glial scar. Molecular biology, immunohistochemistry, spared tissue, and locomotor behavioral studies were used to evaluate astrogliosis, in adult female Sprague-Dawley rats treated with P2 antagonists after moderate injury with the NYU impactor device. Semi-quantitative RT-PCR confirmed the presence of P2Y(1,) P2Y(2,) P2Y(4,) P2Y(6,) P2Y(12), and P2X(2) receptors in the adult spinal cord. Immunohistochemistry studies confirmed a significant decrease in GFAP-labeled cells at the injury epicenter as well as a decrease in spared tissue after treatment with the antagonists. Functional open field testing revealed no significant locomotor score differences between treated and control animals. Our work is consistent with studies suggesting that astrogliosis is an important event after SCI that limits tissue damage and lesion spreading. PMID- 21647707 TI - The birth and early evolution of the frontotemporal dementia (FTD) concept. AB - An historical overview of the development of the concept of frontotemporal dementia is presented, regarding the last 30 years, using as a backbone the conferences held on this theme, with a start in 1986 in Lund, Sweden. Since then, a dramatic increase in research activities and publications has rapidly expanded our knowledge in this field, a step necessary for the ultimate goal to find an effective treatment of this devastating disorder. PMID- 21647708 TI - Role of RhoA/ROCK signaling in endothelial-monocyte-activating polypeptide II opening of the blood-tumor barrier: role of RhoA/ROCK signaling in EMAP II opening of the BTB. AB - The purpose of the present study was to determine the potential for RhoA/ROCK signaling to play a role in endothelial-monocyte-activating polypeptide (EMAP) II induced increase in blood-tumor barrier (BTB) permeability in rat brain microvascular endothelial cells (RBMECs). In the present study, we used an in vitro BTB model, a RhoA inhibitor (C3 exoenzyme) and a ROCK inhibitor (Y27632) to determine whether RhoA/ROCK pathway play a role in the process of TJ disassembly, stress fiber formation, MLC and cofilin phosphorylation, as well as increase of BTB permeability induced by EMAP II. The results revealed that BTB permeability was increased by EMAP II induction, and C3 exoenzyme or Y27632 could partially inhibit the EMAP II-induced increase of BTB permeability. The significant down regulations in tight junction (TJ)-associated proteins occludin, claudin-5 and ZO 1 and stress fiber formation by EMAP II administration were observed, which were partly prevented by C3 exoenzyme or Y27632 pretreatment. Moreover, the significant increases in RhoA activity, myosin light chain (MLC) and cofilin phosphorylation by EMAP II administration were observed, MLC and cofilin phosphorylation were partly inhibited by C3 exoenzyme or Y27632 pretreatment. The present study demonstrates that the activation of RhoA/ROCK signaling in RBMECs was required for the increase of BTB permeability and these effects are related with the ability for RhoA/ROCK to mediate TJ disassembly and stress fiber formation by phosphorylating cofilin and MLC. PMID- 21647709 TI - Activity-dependent neuroprotective protein modulates its own gene expression. AB - We investigated whether activity-dependent neuroprotective protein (ADNP) could autoregulate its own expression. Both the endogenous ADNP gene and reporter gene constructs were analysed in response to overexpression of ADNP, supplied either as wild-type ADNP or a mutant form lacking the NAP motif, a motif which has neuroprotective properties. Overexpression of these two forms of ADNP resulted in both decreased endogenous ADNP expression and repressed ADNP promoter-directed reporter gene activity. Chromatin immunoprecipitation demonstrated the ability of ADNP to bind to its own promoter which is consistent with its action as a repressor of both promoter-supported and endogenous ADNP expression. PMID- 21647710 TI - Gastrointestinal dysfunction in a Parkinson's disease rat model and the changes of dopaminergic, nitric oxidergic, and cholinergic neurotransmitters in myenteric plexus. AB - This study aims to explore the gastrointestinal dysfunction and the changes of dopaminergic, nitric oxidergic, and cholinergic neurons in the myenteric plexus of a Parkinson's disease (PD) rat model. A PD rat model was induced through unilateral substantia nigra administration of 6-hydroxydopamine. Four weeks later, the feces in 1 h and residual solid food in stomach at 2 h after feeding were measured. Changes in tyrosine hydroxylase (TH) in substantial nigra, TH, choline acetyltransferase (ChAT), and neuronal nitric oxide synthase (nNOS) in gastric antrum and colon tissue were examined by immunohistochemistry. Reverse transcription (RT) polymerase chain reaction (PCR) and Western blot were used to evaluate and compare the levels of messenger RNA (mRNA) and protein expression of TH, ChAT, and nNOS in the GI tract between normal and 6-hydroxydopamine-lesioned rats. Compared with control samples, the number of TH+ cells in the damaged side of substantia nigra of 6-hydroxydopamine-lesioned rats decreased significantly (P < 0.01). The weight and water content of the fecal matter decreased (P < 0.01), and the percentage of residual solid food increased (P < 0.01). The average integrated optical densities of TH-positive areas in the gastric antrum and colon tissue increased significantly (P < 0.01), nNOS decreased significantly (P < 0.01), and there were no significant changes in ChAT (P > 0.05). TH and nNOS mRNA levels in the gastric antrum and proximal colon decreased (P < 0.01), there were no significant changes in ChAT mRNA levels (P > 0.05). The protein levels of TH in the GI tract were significantly increased (P < 0.01), nNOS significantly decreased (P < 0.01), and ChAT had no significant changes (P > 0.05). 6 Hydroxydopamine-lesioned rats had delayed gastric emptying and constipation that might be related to the gastrointestinal TH increase and nNOS decrease. These symptoms were not related to changes in cholinergic transmitters. PMID- 21647711 TI - Striatal microinjection of Sydenham chorea antibodies: using a rat model to examine the dopamine hypothesis. AB - We hypothesized that injection of anti-basal antibodies from patients with Sydenham's chorea into rats' striatum will induce behavioral and histological changes. Antibodies from eight Sydenham's chorea patients and eight age-matched controls were injected into the left caudate of 16 rats. Apomorphine- and amphetamine-induced rotations were performed on days 10 and 17, respectively, followed by immunohistochemical studies. Antibodies from patients with Sydenham's chorea, but not controls, bound to a ~50-kDa molecule in the striatum extract; immunohistology staining demonstrated specific binding to cellular component(s) in rats' striatum. Contrary to our hypothesis, we could not detect in the rats injected with Sydenham's chorea antibodies changes in rotational behavior or immunohistochemistry staining for dopaminergic or GABAergic markers. Injection of small quantities of anti-neuronal antibodies present in patients with Sydenham's chorea into rat striatum is insufficient to alter motor behavior or cause detectable cellular changes. PMID- 21647712 TI - Medical management of frontotemporal dementias: the importance of the caregiver in symptom assessment and guidance of treatment strategies. AB - There are no currently Food and Drug Administration-approved or proven off-label treatments for the frontotemporal dementias (FTD). Clinicians, caregivers, and patients struggle regularly to find therapeutic regimens that can alleviate the problematic behavioral and cognitive symptoms associated with these devastating conditions. Success is "hit or miss" and the lessons learned are largely anecdotal to date. Drug discovery in this area has been largely hampered by the heterogeneous clinical presentations and pathological phenotypes of disease that represent significant obstacles to progress in this area. Biologically, plausible treatment strategies include the use of antidepressants (selective serotonin reuptake inhibitors or serotonin-specific reuptake inhibitor and monoamine oxidase inhibitors), acetylcholinesterase inhibitors, N-methyl-D-aspartic acid antagonists, mood stabilizers, antipsychotics, stimulants, antihypertensives, and agents that may ameliorate the symptoms of parkinsonism, pseudobulbar affect, and motor neuron disease that can often coexist with FTD. These medications all carry potential risks as well as possible benefits for the person suffering from FTD, and a clear understanding of these factors is critical in selecting an appropriate therapeutic regimen to maximize cognition and daily functions, reduce behavioral symptoms, and alleviate caregiver burden in an individual patient. The role of the caregiver in tracking and reporting of symptoms and the effects of individual therapeutic interventions is pivotal in this process. This manuscript highlights the importance of establishing an effective therapeutic partnership between the physician and caregiver in the medical management of the person suffering from FTD. PMID- 21647713 TI - Distribution of alarin immunoreactivity in the mouse brain. AB - Alarin is a 25 amino acid peptide that belongs to the galanin peptide family. It is derived from the galanin-like peptide gene by a splice variant, which excludes exon 3. Alarin was first identified in gangliocytes of neuroblastic tumors and later shown to have a vasoactive function in the skin. Recently, alarin was demonstrated to stimulate food intake as well as the hypothalamic-pituitary gonadal axis in rodents, suggesting that it might be a neuromodulatory peptide in the brain. However, the individual neurons in the central nervous system that express alarin have not been identified. Here, we determined the distribution of alarin-like immunoreactivity (alarin-LI) in the adult murine brain. The specificity of the antibody against alarin was demonstrated by the absence of labeling after pre-absorption of the antiserum with synthetic alarin peptide and in transgenic mouse brains lacking neurons expressing the GALP gene. Alarin-LI was observed in different areas of the murine brain. A high intensity of alarin LI was detected in the accessory olfactory bulb, the medial preoptic area, the amygdala, different nuclei of the hypothalamus such as the arcuate nucleus and the ventromedial hypothalamic nucleus, the trigeminal complex, the locus coeruleus, the ventral chochlear nucleus, the facial nucleus, and the epithelial layer of the plexus choroideus. The distinct expression pattern of alarin in the adult mouse brain suggests potential functions in reproduction and metabolism. PMID- 21647715 TI - Drug use patterns for the prevention of paediatric low cardiac output syndrome in Europe. PMID- 21647716 TI - Prognostic models for predicting mortality in elderly ICU patients: a systematic review. AB - PURPOSE: To systematically review prognostic research literature on development and/or validation of mortality predictive models in elderly patients. METHODS: We searched the Scopus database until June 2010 for articles aimed at validating prognostic models for survival or mortality in elderly intensive care unit (ICU) patients. We assessed the models' fitness for their intended purpose on the basis of barriers for use reported in the literature, using the following categories: (1) clinical credibility, (2) methodological quality (based on an existing quality assessment framework), (3) external validity, (4) model performance, and (5) clinical effectiveness. RESULTS: Seven studies were identified which met our inclusion criteria, one of which was an external validation study. In total, 17 models were found of which six were developed for the general adult ICU population and eleven specifically for elderly patients. Cohorts ranged from 148 to 12,993 patients and only smaller ones were obtained prospectively. The area under the receiver operating characteristic curve (AUC) was most commonly used to measure performance (range 0.71-0.88). The median number of criteria met for clinical credibility was 4.5 out of 7 (range 2.5-5.5) and 17 out of 20 for methodological quality (range 15-20). CONCLUSIONS: Although the models scored relatively well on methodological quality, none of them can be currently considered sufficiently credible or valid to be applicable in clinical practice for elderly patients. Future research should focus on external validation, addressing performance measures relevant for their intended use, and on clinical credibility including the incorporation of factors specific for the elderly population. PMID- 21647717 TI - Brain temperature in volunteers subjected to intranasal cooling. AB - PURPOSE: Intranasal cooling can be used to initiate therapeutic hypothermia. However, direct measurement of brain temperature is difficult and the intra cerebral distribution of temperature changes with cooling is unknown. The purpose of this study was to measure the brain temperature of human volunteers subjected to intranasal cooling using non-invasive magnetic resonance (MR) methods. METHODS: Intranasal balloons catheters circulated with saline at 20 degrees C were applied for 60 min in ten awake volunteers. No sedation was used. Brain temperature changes were measured and mapped using MR spectroscopic imaging (MRSI) and phase-mapping techniques. Heart rate and blood pressure were monitored throughout the experiment. Rectal temperature was measured before and after the cooling. Mini Mental State Examination (MMSE) test and nasal inspection were done before and after the cooling. Questionnaires about the subjects' personal experience were completed after the experiment. RESULTS: Brain temperature decrease measured by MRSI was -1.7 +/- 0.8 degrees C and by phase-mapping -1.8 +/ 0.9 degrees C (n = 9) at the end of cooling. Spatial distribution of temperature changes was relatively uniform. Rectal temperature decreased by -0.5 +/- 0.3 degrees C (n = 5). The physiological parameters were stable and no shivering was reported. The volunteers remained alert during cooling and no cognitive dysfunctions were apparent in the MMSE test. Postcooling nasal examination detected increased nasal secretion in nine of the ten volunteers. Volunteers' acceptance of the method was good. CONCLUSION: Both MR techniques revealed brain temperature reductions after 60 min of intranasal cooling with balloons circulated with saline at 20 degrees C in awake, unsedated volunteers. PMID- 21647718 TI - End-inspiratory rebreathing reduces the end-tidal to arterial PCO2 gradient in mechanically ventilated pigs. AB - PURPOSE: Noninvasive monitoring of the arterial partial pressures of CO(2) (PaCO(2)) of critically ill patients by measuring their end-tidal partial pressures of CO(2) (PETCO(2)) would be of great clinical value. However, the gradient between PETCO(2) and PaCO(2) (PET-aCO(2)) in such patients typically varies over a wide range. A reduction of the PET-aCO(2) gradient can be achieved in spontaneously breathing healthy humans using an end-inspiratory rebreathing technique. We investigated whether this method would be effective in reducing the PET-aCO(2) gradient in a ventilated animal model. METHODS: Six anesthetized pigs were ventilated mechanically. End-tidal gases were systematically adjusted over a wide range of PETCO(2) (30-55 mmHg) and PETO(2) (35-500 mmHg) while employing the end-inspiratory rebreathing technique and measuring the PET-aCO(2) gradient. Duplicate arterial blood samples were taken for blood gas analysis at each set of gas tensions. RESULTS: PETCO(2) and PaCO(2) remained equal within the error of measurement at all gas tension combinations. The mean +/- SD PET-aCO(2) gradient (0.13 +/- 0.12 mmHg, 95% CI -0.36, 0.10) was the same (p = 0.66) as that between duplicate PaCO(2) measurements at all PETCO(2) and PETO(2) combinations (0.19 +/- 0.06, 95% CI -0.32, -0.06). CONCLUSIONS: The end-inspiratory rebreathing technique is capable of reducing the PET-aCO(2) gradient sufficiently to make the noninvasive measurement of PETCO(2) a useful clinical surrogate for PaCO(2) over a wide range of PETCO(2) and PETO(2) combinations in mechanically ventilated pigs. Further studies in the presence of severe ventilation-perfusion (V/Q) mismatching will be required to identify the limitations of the method. PMID- 21647719 TI - Rule of rescue or the good of the many? An analysis of physicians' and nurses' preferences for allocating ICU beds. AB - PURPOSE: To examine intensive care unit (ICU) clinicians' willingness to trade off societal benefits in favor of a small chance of rescuing an identifiable critically ill patient. METHODS: We sent mixed-methods questionnaires to national samples of US ICU clinicians, soliciting their preferences for allocating their last bed to a gravely ill patient with little chance to survive, versus a deceased or dying patient for whom aggressive management could help others through organ donation. RESULTS: Complete responses were obtained from 684 of 2,206 physicians (31.0%) and 438 of 988 nurses (44.3%); there was no evidence of non-response bias. Physicians were more likely than nurses to adhere to the "rule of rescue" by allocating the last bed to the gravely ill patient (45.9 vs. 32.6%, difference = 13.2%; 95% CI 9.1-17.3%). The magnitude of the social benefit to be obtained through organ donor management (5 or 30 life-years added for transplant recipients) had small and inconsistent effects on clinicians' willingness to prioritize the donor. In qualitative analyses, the most common reason for allocating the last bed to an identifiable patient (identified by 65% of physicians and 75% of nurses) was that clinicians perceived strong obligations to identifiable living patients. CONCLUSIONS: More than one-third of ICU clinicians forewent substantial social benefits so as to devote resources to an individual patient unlikely to benefit from them. Such allegiance to the rule of rescue suggests challenges for efforts to reform ICU triage practices. PMID- 21647720 TI - Impact of hypoxic hepatitis on mortality in the intensive care unit. AB - PURPOSE: Hypoxic hepatitis (HH) is a form of hepatic injury following arterial hypoxemia, ischemia, and passive congestion of the liver. We investigated the incidence and the prognostic implications of HH in the medical intensive care unit (ICU). METHODS: A total of 1,066 consecutive ICU admissions at three medical ICUs of a university hospital were included in this prospective cohort study. All patients were screened prospectively for the presence of HH according to established criteria. Independent risk factors of mortality in this cohort of critically ill patients were identified by a multivariate Poisson regression model. RESULTS: A total of 118 admissions (11%) had HH during their ICU stay. These patients had different baseline characteristics, longer median ICU stay (8 vs. 6 days, p < 0.001), and decreased ICU survival (43 vs. 83%, p < 0.001). The crude mortality rate ratio of admissions with HH was 4.62 (95% CI 3.63-5.86, p < 0.001). Regression analysis demonstrated strong mortality risk for admissions with HH requiring vasopressor therapy (adjusted rate ratio 4.91; 95% CI 2.51 9.60, p < 0.001), whereas HH was not significantly associated with mortality in admissions without vasopressor therapy (adjusted rate ratio 1.79, 95% CI 0.52 6.23, p = 0.359). CONCLUSIONS: Hypoxic hepatitis (HH) occurs frequently in the medical ICU. The presence of HH is a strong risk factor for mortality in the ICU in patients requiring vasopressor therapy. PMID- 21647721 TI - Adult patients are more catabolic than children during acute phase after burn injury: a retrospective analysis on muscle protein kinetics. AB - PURPOSE: This study was performed to determine if there is an age-related specificity in the response of muscle protein metabolism to severe burn injury during acute hospitalization. This is a retrospective analysis of previously published data. METHODS: Nineteen adult and 58 pediatric burn-injured patients (age 43.3 +/- 14.3 vs. 7.2 +/- 5.3 years, adult vs. children) participated in stable isotope [ring-(2)H(5)]phenylalanine (Phe) infusion studies. Femoral arterial and venous blood samples and muscle biopsy samples were collected throughout the study. Data are presented as means +/- standard deviation (SD). A p value less than 0.05 was considered statistically significant. RESULTS: Muscle net protein balance (NB) was higher in children (adult vs. children, -43 +/- 61 vs. 8 +/- 68 nmol Phe/min/100 ml leg volume, p < 0.05). Muscle protein fractional synthesis rate (FSR) was higher in children (adult vs. children, 0.11 +/- 0.05 vs. 0.16 +/- 0.10 %/h, p < 0.05). Leg muscle protein breakdown was not different between the groups (adult vs. children, 179 +/- 115 vs. 184 +/- 124 nmol Phe/min/100 ml leg volume, p > 0.05); synthesis rate was 134 +/- 96 and 192 +/- 128 nmol Phe/min/100 ml leg volume in adults and children, respectively (p = 0.07). Age significantly correlated with muscle protein NB (p = 0.01) and FSR (p = 0.02); but not with breakdown (p = 0.67) and synthesis (p = 0.07) rates measured by using a three-pool model. CONCLUSION: In burn injury, the muscle protein breakdown may be affected to the same extent in adults and children, whereas synthesis may have age-related specificities, resulting in a better but still low NB in children. PMID- 21647722 TI - Polar metabolites synergize the activity of prostaglandin F2alpha in a species specific hormonal sex pheromone released by ovulated common carp. AB - Many species of teleost fish detect and release F prostaglandins (PGFs), but the specific identities of these compounds and how they function as species-specific pheromones have yet to be resolved. This study addressed these questions in the common carp. An initial set of experiments established that mature male common carp were attracted to chemicals released by ovulated conspecifics, whereas the odor of female goldfish, a close relative, was less attractive. Tests of fractionated holding water from ovulated carp revealed that only the non-polar fraction was attractive on its own. Mass spectrometry and immunoassay next demonstrated that the non-polar fraction contained large quantities of prostaglandin F(2alpha) (PGF(2alpha)), 15keto-prostaglandinF(2alpha), and 13,14 dihydro-15keto-prostaglandin F(2alpha) (100 g fish released over 1 MUg of all 3 PGFs per h at a ratio of 1.0: 1.7: 0.7). Ovulated goldfish released the same three PGFs but at a slightly greater rate and in a different ratio. Tests of synthetic mixtures of these PGFs revealed that the carp-specific mixture attracted male carp but was no better than the goldfish-specific mixture or PGF(2alpha) alone and that PGF(2alpha) was just as attractive as mixture of all three PGFs. A final set of attraction tests revealed that although PGF(2alpha) could explain all of the activity of the non-polar portion of female carp holding water, it could not explain the entire activity of female water but that a mixture of PGFs and the polar fraction could. We conclude that ovulated female carp release a multi-component sex pheromone complex that is comprised of PGF(2alpha) and unknown species-specific polar compound(s) that synergize the activity of the former. The pheromone also might be useful in controlling this invasive species. The observation that a fish hormonal sex pheromone incorporates bodily metabolites in addition to relatively common hormonal products demonstrates a mechanism by which species specificity may be conferred to this common type of sex pheromone. PMID- 21647723 TI - Searching for major urinary proteins (MUPs) as chemosignals in urine of subterranean rodents. AB - Chemosensory information mediates behavior in many rodent genera. Major Urinary Proteins (MUPs) facilitate chemical communication in some species of mice. We sought to demonstrate the importance of MUPs in chemosignaling across a range of rodent genera that live in different habitats and social structures. We analyzed urine from three subterranean rodent genera from different continents, and with diverse social systems: eusocial Zambian mole-rats (Fukomys), solitary Israeli blind mole rats (Spalax), and social Chilean coruros (Spalacopus). 2D gel electrophoresis revealed low levels of protein, with sequences similar to aphrodisin, in Fukomys mole-rat urine, but no MUPs in urine of any of the studied species. Previous research demonstrated that subjects from the tested genera responded differentially to odors indicating transmission of individuality, family/colony or population, species, and reproductive state in secretions and excretions of conspecifics. This extends, to subterranean rodents, the evidence that rodent species can successfully transmit and receive chemosignals without the necessity of MUPs. PMID- 21647724 TI - Long-term effectiveness of a back education programme in elementary schoolchildren: an 8-year follow-up study. AB - The purpose of this study was to investigate the long-term effectiveness of a spine care education programme conducted in 9- to 11-year-old schoolchildren. The study sample included 96 intervention subjects and 98 controls (9- to 11-year olds at baseline). Intervention consisted of a 6-week school-based back education programme (predominantly biomechanically oriented) and was implemented by a physical therapist. Self-reported outcomes on back care knowledge, spinal care behaviour, self-efficacy towards favourable back care behaviour, prevalence of back and neck pain during the week and fear-avoidance beliefs were evaluated by the use of questionnaires. Post-tests were performed within 1 week after programme completion, after 1 year and after 8 years. Whereas the educational back care programme resulted in increased back care knowledge up to adulthood (P < 0.001), intervention did not change spinal care behaviour or self-efficacy. Pain prevalence figures increased less in the experimental group compared to the controls over the 8-year time span, yet statistical significance was not reached. Dropout analysis revealed spinal pain prevalence rates to be different in both groups throughout the study, including at baseline. Back education at young age did not reinforce fear-avoidance beliefs up to adulthood. Predominantly biomechanical oriented back education in elementary schoolchildren is effective in improving the cognitive aspect of back care up to adulthood, yet not in changing actual behaviour or self-efficacy. The current study does not provide evidence that educational back care programmes have any impact on spinal pain in adulthood. The true long-term impact of school-based spinal health interventions on clinically relevant outcome measures merits further attention. PMID- 21647726 TI - Dramatic response to zonisamide of post-subarachnoid hemorrhage Holmes' tremor. PMID- 21647725 TI - Dysphagia in patients with acute striatocapsular hemorrhage. AB - Dysphagia is found in up to 80% of acute stroke patients. To date most studies have focused on ischemic stroke only. Little is known about the incidence and pattern of dysphagia in hemorrhagic stroke. Here we describe the characteristics of dysphagia in patients with striatocapsular hemorrhage. Fiberoptic Endoscopic Evaluation of Swallowing (FEES) was carried out in 30 patients with acute striatocapsular hemorrhage. Dysphagia was classified according to the six-point Fiberoptic Endoscopic Dysphagia Severity Scale (FEDSS) within 72 h after admission. Lesion volume, hemisphere and occurrence of ventricular rupture were determined from computer tomography scans. Data on initial NIH-SS, clinical symptoms, need for endotracheal intubation, diagnosis of pneumonia and feeding status on discharge were recorded. Swallowing impairment was observed in 76.7% of patients (n = 23). Mean FEDSS score was 3.1 +/- 1.5. Main findings were penetration or aspiration of liquids as well as leakage to valleculae and piriform sinus. Incidence of pneumonia was 30.0% (n = 9). Age, NIH-SS and hematoma volume did not correlate with dysphagia severity. None of the clinical characteristics was predictive for dysphagia. On discharge after 12.9 +/- 5.3 days, a two-point improvement on the FEDSS was seen in seven patients, (30.4%) and five patients (21.7%) had gained at least one point. In striatocapsular hemorrhage, dysphagia is a common and so far underrecognized symptom. FEES results indicate predominant impairment of oral motor control. Swallowing impairment is not related to other clinical deficits, stroke severity or lesion characteristics. Thus, detailed dysphagia assessment is indicated in all cases. PMID- 21647727 TI - Stiff person syndrome (SPS) complicated by respiratory failure: successful treatment with rituximab. PMID- 21647728 TI - A novel protein S gene mutation combined with protein S Tokushima mutation in a patient with superior sagittal sinus thrombosis. PMID- 21647729 TI - Levodopa-induced breathing, cognitive and behavioral changes in Parkinson's disease. PMID- 21647730 TI - Progressive multifocal leukoencephalopathy and natalizumab. AB - Natalizumab (TYSABRI((r))), a specific alpha4-integrin antagonist, is approved as a second-line treatment of relapsing-remitting MS (RRMS) patients who fail therapy with interferons or as first-line treatment of patients with highly active relapsing-remitting disease. Since the market introduction of natalizumab as a monotherapy in July of 2006, 111 cases of PML have been reported in natalizumab-treated MS patients as of April 2011. This review focuses on the available data regarding risk stratification for PML under long-term natalizumab therapy, and summarizes the current approach for PML management, as a natalizumab treatment complication is not necessarily associated with a fatal outcome. There is a need for development of surrogate markers that would help to better define the risk of PML in individual patients. PMID- 21647731 TI - Diversity and bioethics. PMID- 21647732 TI - The current status of decision-making procedures and quality assurance in Europe: an overview. AB - The 2005 Report on Social Responsibility and Health of the UNESCO International Bioethics Committee (Ibc) proposes a new approach to implementing the right to healthcare and suggests a number of Courses of Action to be followed in various fields. Based on the latest available data, we intend to present an overview of the current state of European health systems in two of those fields-decision making procedures and quality assurance in health care-and to attempt a comparison of the situation with the Report's provisions, in order to pave the way for the identification of what still has to be done to bridge international recommendations and the reality of policy and practice in Europe's health care. PMID- 21647734 TI - Prevalence of dysplasia as the source of worse outcome in young female patients after hip resurfacing arthroplasty. AB - PURPOSE: Smaller femoral component size has been implicated as underlying the risk factor that explains the higher failure rate in women who have a hip resurfacing arthroplasty (HRA). We suspect that the diagnosis of dysplasia may be a more important causative risk factor than either small component size or female gender. METHODS: From January 2002 to July 2008, a total of 1,216 HRA cases, 1,082 with the primary diagnosis of osteoarthritis and 134 with dysplasia, were included in this study. Of them, 867 cases were performed in men and 349 performed in women. The average femoral component size was 51 +/- 4 mm. Cox proportional hazard regression models were used to evaluate the significance of each variable and determine the causative risk factors for failure. RESULTS: The average follow-up was 5 +/- 2 years. Thirty-nine cases failed (20 in men vs. 19 in women). The failure rate for the whole group was 3.2% (2.3% in men vs. 5.4% in women; P = 0.01). Dysplasia (P = 0.05) was identified as the only significant risk factor in our multi-variable analysis; small femoral component size (P = 0.09) and gender (P = 0.76) were not significant risk factors. Women with the primary diagnosis of dysplasia had a survivorship rate of only 75% compared to 93% for the entire group at eight-year follow-up post-operatively. CONCLUSIONS: In our study, we found that the high incidence of dysplasia in young women undergoing HRA was the reason that women had a higher failure rate after HRA. In dysplasia, 70% of failures were due to acetabular problems, of which 50% were due to failure of fixation and 20% due to adverse wear. PMID- 21647735 TI - Influence of total knee arthroplasty on patellar kinematics and contact characteristics. AB - PURPOSE: Femoro-patellar complications are one of the most common problems after total knee arthroplasty (TKA). TKA components that reduce patellar loads and preserve physiological patellar kinematics should reduce these problems. Therefore, we evaluated the patellar kinematics and the retro-patellar contact characteristics in both the intact knee and in the TKA-knee. METHODS: Eight Thiel embalmed cadaver knees were tested first intact and then after TKA using rotating as well as gliding inlay and with additional patellar resurfacing while flexing the knee from 0 degrees to 100 degrees . During the examination quadriceps and hamstring forces were applied. RESULTS: TKA with additional patellar resurfacing led to an increased retro-patellar pressure, a decreased contact area and an increased lateral movement. Although patellar kinematics could not be changed by using a gliding inlay compared to a rotating inlay, the gliding inlay improved retro-patellar contact characteristics by reducing the pressure and increasing the contact area, especially in higher flexion. CONCLUSIONS: The increased retro patellar pressure together with the increased lateral movement of the patella after TKA may be one important cause for anterior knee pain appearing after TKA. In view of the improved retro-patellar contact characteristics using a gliding inlay this inlay should be preferred, providing that the posterior cruciate ligament is intact. PMID- 21647736 TI - Tibial lengthening over an intramedullary nail in patients with short stature or leg-length discrepancy: a comparative study. AB - PURPOSE: The aim of this study was to review our experiences with tibial lengthening over an intramedullary nail in comparison to the conventional Ilizarov method. METHODS: We performed a retrospective comparison of tibial lengthening using the conventional Ilizarov method (group A: 23 limbs in 13 patients) versus over a nail (group B: 51 limbs in 26 patients). The percentage increase in tibial length, lengthening index, external fixation index, consolidation index and complications were assessed. RESULTS: The mean gain in tibial length was 7.4 cm, which represents a mean increase of 26.0%. There was no difference in lengthening index or consolidation index; however, the patients in group A wore the external fixator longer than those in group B (281.5 versus 129.0 days), which represents a larger external fixation index (40.0 versus 17.4 day/cm). Group A had a higher complication rate (1.0 versus 0.47 per tibia) than group B. CONCLUSIONS: Tibial lengthening over an intramedullary nail confers advantages over the conventional Ilizarov method, including shorter time needed for external fixation and lower complication rates. PMID- 21647737 TI - iScreen: world's first cloud-computing web server for virtual screening and de novo drug design based on TCM database@Taiwan. AB - The rapidly advancing researches on traditional Chinese medicine (TCM) have greatly intrigued pharmaceutical industries worldwide. To take initiative in the next generation of drug development, we constructed a cloud-computing system for TCM intelligent screening system (iScreen) based on TCM Database@Taiwan. iScreen is compacted web server for TCM docking and followed by customized de novo drug design. We further implemented a protein preparation tool that both extract protein of interest from a raw input file and estimate the size of ligand bind site. In addition, iScreen is designed in user-friendly graphic interface for users who have less experience with the command line systems. For customized docking, multiple docking services, including standard, in-water, pH environment, and flexible docking modes are implemented. Users can download first 200 TCM compounds of best docking results. For TCM de novo drug design, iScreen provides multiple molecular descriptors for a user's interest. iScreen is the world's first web server that employs world's largest TCM database for virtual screening and de novo drug design. We believe our web server can lead TCM research to a new era of drug development. The TCM docking and screening server is available at http://iScreen.cmu.edu.tw/. PMID- 21647738 TI - Genome-wide association with C-reactive protein levels in CLHNS: evidence for the CRP and HNF1A loci and their interaction with exposure to a pathogenic environment. AB - Recent genome-wide association studies have related several genetic loci, including C-reactive protein (CRP), hepatocyte nuclear factor 1 homeobox (HNF1A), and genetic variations in the leptin receptor (LEPR), to circulating CRP levels in populations of European ancestry. The genetic effects in other populations and across varying levels of exposure to a pathogenic environment, an important environmental factor associated with CRP, remain to be determined. We tested 2,073,674 single-nucleotide polymorphisms (SNPs) for association with plasma CRP (limited to <=10 mg/L) in 1,709 unrelated Filipino women from the Cebu Longitudinal Health and Nutrition Survey. The strongest evidence of association was observed with variants at CRP (rs876537, P = 1.4 * 10(-9)) and HNF1A (rs7305618, P = 1.0 * 10(-8)). Among other previously reported CRP-associated loci, the apolipoprotein E epsilon4 haplotype was associated with decreased CRP level (P = 7.1 * 10(-4)), and modest association was observed with LEPR (rs1892534, P = 0.076), with direction of effects consistent with previous studies. The strongest signal at a locus not previously reported mapped to a gene desert region on chromosome 6q16.1 (rs1408282, P = 2.9 * 10(-6)). Finally, we observed nominal evidence of interaction with exposure to a pathogenic environment for top main effect SNPs at HNF1A (rs7305618, P = 0.031), LEPR (rs1892535, P = 0.030) and 6q16.1 (rs1408282, P = 0.046). Our findings demonstrate convincing evidence that genetic variants in CRP and HNF1A contribute to plasma CRP in Filipino women and provide the first evidence that exposure to a pathogenic environment may modify the genetic influence at the HNF1A, LEPR, and 6q16.1 loci on plasma CRP level. PMID- 21647739 TI - Relapse of posttraumatic osteomyelitis due to Clostridium celerecrescens. PMID- 21647740 TI - Temporal dynamics in the evolution of the sunflower genome as revealed by sequencing and annotation of three large genomic regions. AB - Improved knowledge of genome composition, especially of its repetitive component, generates important informations in both theoretical and applied research. In this study, we provide the first insight into the local organization of the sunflower genome by sequencing and annotating 349,380 bp from 3 BAC clones, each including one single-copy gene. These analyses resulted in the identification of 11 putative gene sequences, 18 full-length LTR retrotransposons, 6 incomplete LTR retrotransposons, 2 non-autonomous LTR-retroelements (LINEs), 2 putative DNA transposons fragments and one putative helitron. Among LTR-retrotransposons, non autonomous elements (the so-called LARDs), which do not carry any protein encoding sequence, were discovered for the first time in the sunflower. The insertion time of intact retroelements was measured, based on sister LTRs divergence. All isolated elements were inserted relatively recently, especially those belonging to the Gypsy superfamily. Retrotransposon families related to those identified in the BAC clones are present also in other species of Helianthus, both annual and perennial, and even in other Asteraceae. In one of the three BAC clones, we found five copies of a lipid transfer protein (LTP) encoding gene within less than 100,000 bp, four of which are potentially functional. Two of these are interrupted by LTR retrotransposons, in the intron and in the coding sequence, respectively. The divergence between sister LTRs of the retrotransposons inserted within the genes indicates that LTP gene duplication started earlier than 1.749 MYRS ago. On the whole, the results reported in this study confirm that the sunflower is an excellent system to study transposons dynamics and evolution. PMID- 21647741 TI - Improved accuracy in measuring one-bond and two-bond (15)N, (13)C (alpha) coupling constants in proteins by double-inphase/antiphase (DIPAP) spectroscopy. AB - An extension to HN(CO-alpha/beta-N,C(alpha)-J)-TROSY (Permi and Annila in J Biomol NMR 16:221-227, 2000) is proposed that permits the simultaneous determination of the four coupling constants (1) J (N'(i)Calpha(i)), (2) J (HN(i)Calpha(i)), (2) J (Calpha(i-1)N'(i)), and (3) J (Calpha(i-1)HN(i)) in (15)N,(13)C-labeled proteins. Contrasting the original scheme, in which two separate subspectra exhibit the (2) J (CalphaN') coupling as inphase and antiphase splitting (IPAP), we here record four subspectra that exhibit all combinations of inphase and antiphase splittings possible with respect to both (2) J (CalphaN') and (1) J (N'Calpha) (DIPAP). Complementary sign patterns in the different spectrum constituents overdetermine the coupling constants which can thus be extracted at higher accuracy than is possible with the original experiment. Fully exploiting data redundance, simultaneous 2D lineshape fitting of the E.COSY multiplet tilts in all four subspectra provides all coupling constants at ultimate precision. Cross-correlation and differential-relaxation effects were taken into account in the evaluation procedure. By applying a four point Fourier transform, the set of spectra is reversibly interconverted between DIPAP and spin-state representations. Methods are exemplified using proteins of various size. PMID- 21647742 TI - The effect of chemotherapy on expression of folate receptor-alpha in ovarian cancer. AB - BACKGROUND: Folate receptor alpha (FR-alpha) has been identified as a potential target in ovarian cancer for diagnostic and therapeutic purposes, based on its overexpression in serous epithelial ovarian carcinoma. The effect of chemotherapy on FR-alpha expression may be important in the applicability of FR-alpha directed agents in the case of residual tumor tissue. The objective of this study was to assess FR-alpha expression in ovarian carcinoma and to evaluate whether FR-alpha expression is altered by chemotherapy. MATERIALS & METHODS: FR-alpha expression was analyzed by semi-quantitative scoring of immunohistochemical staining on tissue microarrays (TMAs) from a database containing 361 ovarian cancer tissue samples, of which 210 serous and 116 non-serous carcinoma (35 missing). Serous carcinoma samples included 28 matched samples with tissue from both primary surgery and interval debulking surgery, and 12 matched samples with tissue from both primary surgery and surgery for recurrent disease. RESULTS: FR-alpha expression was seen in 81.8% of serous ovarian cancers versus 39.9% of non-serous carcinomas (p < 0.001). In matched serous carcinoma samples, no significant change in FR-alpha expression in vital tumor tissue after chemotherapy was observed (p = 0.1). FR-alpha expression was not a prognostic marker of progression free survival (p = 0.8) or overall survival (p = 0.7). CONCLUSION: FR alpha was expressed in the majority of serous ovarian tumors, although >50% of cases showed only weak expression. Chemotherapy did not alter expression rates in remaining vital tumor tissue, indicating that folate-targeted agents may have a place in the treatment for ovarian cancer, before as well as after chemotherapy. Furthermore, FR-alpha status did not influence survival. PMID- 21647743 TI - [Pregabalin and gabapentin in multiple sclerosis: clinical experiences and therapeutic implications]. AB - BACKGROUND: Due to a plethora of additional symptoms patients with multiple sclerosis (MS) receive symptomatic treatment besides disease-modifying therapies. Among the substances which are commonly used are ion channel modulators (e. g. pregabalin, gabapentin, carbamazepine). The aim of this study was to investigate the use of these drugs in clinical practice in a larger patient cohort. PATIENTS: Data from 533 MS patients [439 without and 94 patients with add-on therapy (treatment group)] were evaluated retrospectively. All patients received a detailed neurological examination including evaluation of EDSS scores. RESULTS: Pregabalin and gabapentin are used most commonly. Abnormal sensations are the most frequent reason for therapy initiation. Patients with higher EDSS values and/or under mitoxantrone treatment most frequently receive additional therapy. CONCLUSION: So far, it is not known whether the investigated agents exert a beneficial influence on the disease course of MS itself beyond a mere symptomatic treatment. Further research efforts and clinical studies are necessary to address this question. PMID- 21647744 TI - [Hirayama disease in Germany: case reports and review of the literature]. AB - Hirayama disease is a juvenile benign distal upper limb muscular atrophy rarely observed in Europe, usually monomelic involving C7-Th1 innervated muscles. It is characterized by insidious onset and a self-limited course within a few years. The pathogenesis of this mostly sporadic disease is not fully clarified. Cervical flexion myelopathy with mechanical ischemic damage of spinal motoneurons is the best established pathogenetic hypothesis, but neurodegenerative and autoimmune causes are also debated. Typically, young men of Asian origin are affected. Here we describe three German Caucasian patients with Hirayama disease and provide an up-to-date review of the literature. PMID- 21647745 TI - Optimizing diabetes self-care in low literacy and minority populations--problem solving, empowerment, peer support and technology-based approaches. PMID- 21647746 TI - Effect of aspirin dose on mortality and cardiovascular events in people with diabetes: a meta-analysis. AB - OBJECTIVES: Pharmacologic evidence suggests adequate antiplatelet activity in diabetic patients requires >100 mg aspirin daily, yet recent trials have used <=100 mg daily. This meta-analysis explored the relationship between aspirin dose and prevention of cardiovascular events. DATA SOURCES: Six electronic databases were searched using database-appropriate terms for aspirin, diabetes, and comparative study from inception until February 2010. REVIEW METHODS: Randomized controlled trials and cohort studies comparing aspirin to no antiplatelet therapy were included if they reported cardiovascular events as pre-specified outcomes, aspirin dose, and number of diabetic patients. Studies were stratified by daily aspirin dose (<=100 mg; 101-325 mg; >325 mg) and pooled risk ratios (RR) were calculated using random effects models. All-cause mortality was the primary outcome of interest. Cardiovascular-related mortality, myocardial infarction, and stroke were secondary outcomes. RESULTS: Data for diabetic patients were available from 21 studies (n = 17,522). Overall, 1,172 (15.4%) of 7,592 aspirin users and 1,520 (18.4%) of 8,269 controls died (p = 0.31). The pooled RRs were 0.89 (95% CI: 0.72-1.10; p = 0.27) from 13 studies using <=100 mg (I(2) = 64%); 0.89 (95% CI: 0.61-1.30; p = 0.55) from four studies using 101-325 mg (I(2) = 83%); and 0.96 (95% CI: 0.85-1.08; p = 0.50) from eight studies using >325 mg (I(2) = 0%). Aspirin use was associated with a significantly lower risk of mortality (RR: 0.82; 95% CI: 0.69-0.98; p = 0.03) in 13 secondary prevention studies (I(2) = 27%), whereas aspirin use in seven primary prevention studies (I(2) = 0%) was not (RR: 1.01; 95% CI 0.85-1.19; p = 0.94). A substantial amount of heterogeneity was observed amongst studies in all outcomes. Although inclusion of cohort studies was a major source of heterogeneity, stratification by study design did not reveal a significant dose-response relationship. CONCLUSIONS/INTERPRETATION: This summary of available data does not support an aspirin dose-response effect for prevention of cardiovascular events in diabetic patients. However, the systematic review identified an important gap in randomized controlled trial evidence for using 101-325 mg aspirin daily in diabetes. PMID- 21647747 TI - From the editors' desk: physician heal thyself. PMID- 21647748 TI - Use of an electronic patient portal among disadvantaged populations. AB - BACKGROUND: Electronic patient portals give patients access to information from their electronic health record and the ability to message their providers. These tools are becoming more widely used and are expected to promote patient engagement with health care. OBJECTIVE: To quantify portal usage and explore potential differences in adoption and use according to patients' socioeconomic and clinical characteristics in a network of federally qualified health centers serving New York City and neighboring counties. DESIGN: Retrospective analysis of data from portal and electronic health records. PARTICIPANTS: 74,368 adult patients seen between April 2008 and April 2010. MAIN MEASURES: Odds of receiving an access code to the portal, activating the account, and using the portal more than once KEY RESULTS: Over the 2 years of the study, 16% of patients (n = 11,903) received an access code. Of these, 60% (n = 7138) activated the account, and 49% (n = 5791) used the account two or more times. Patients with chronic conditions were more likely to receive an access code and to become repeat users. In addition, the odds of receiving an access code were significantly higher for whites, women, younger patients, English speakers, and the insured. The odds of repeat portal use, among those with activated accounts, increased with white race, English language, and private insurance or Medicaid compared to no insurance. Racial disparities were small but persisted in models that controlled for language, insurance, and health status. CONCLUSIONS: We found good early rates of adoption and use of an electronic patient portal during the first 2 years of its deployment among a predominantly low-income population, especially among patients with chronic diseases. Disparities in access to and usage of the portal were evident but were smaller than those reported recently in other populations. Continued efforts will be needed to ensure that portals are usable for and used by disadvantaged groups so that all patients benefit equally from these technologies. PMID- 21647749 TI - Addressing racial healthcare disparities: how can we shift the focus from patients to providers? PMID- 21647750 TI - Reducing barriers to mental health and social services for Iraq and Afghanistan veterans: outcomes of an integrated primary care clinic. AB - BACKGROUND: Despite high rates of post-deployment psychosocial problems in Iraq and Afghanistan veterans, mental health and social services are under-utilized. OBJECTIVE: To evaluate whether a Department of Veterans Affairs (VA) integrated care (IC) clinic (established in April 2007), offering an initial three-part primary care, mental health and social services visit, improved psychosocial services utilization in Iraq and Afghanistan veterans compared to usual care (UC), a standard primary care visit with referral for psychosocial services as needed. DESIGN: Retrospective cohort study using VA administrative data. POPULATION: Five hundred and twenty-six Iraq and Afghanistan veterans initiating primary care at a VA medical center between April 1, 2005 and April 31, 2009. MAIN MEASURES: Multivariable models compared the independent effects of primary care clinic type (IC versus UC) on mental health and social services utilization outcomes. KEY RESULTS: After 2007, compared to UC, veterans presenting to the IC primary care clinic were significantly more likely to have had a within-30-day mental health evaluation (92% versus 59%, p < 0.001) and social services evaluation [77% (IC) versus 56% (UC), p < 0.001]. This exceeded background system wide increases in mental health services utilization that occurred in the UC Clinic after 2007 compared to before 2007. In particular, female veterans, younger veterans, and those with positive mental health screens were independently more likely to have had mental health and social service evaluations if seen in the IC versus UC clinic. Among veterans who screened positive for >= 1 mental health disorder(s), there was a median of 1 follow-up specialty mental health visit within the first year in both clinics. CONCLUSIONS: Among Iraq and Afghanistan veterans new to primary care, an integrated primary care visit further improved the likelihood of an initial mental health and social services evaluation over background increases, but did not improve retention in specialty mental health services. PMID- 21647751 TI - Dermatologic manifestations as indicators of immune status in HIV/AIDS. PMID- 21647752 TI - The correlation of serum trace elements and heavy metals with carotid artery atherosclerosis in maintenance hemodialysis patients. AB - Changes in essential trace elements and heavy metals may affect the atherosclerotic state of patients on maintenance hemodialysis (HD). The aim of the study was to evaluate the relation between the serum levels of some trace elements and heavy metals (iron, zinc, manganese, copper, magnesium, cobalt, cadmium, lead, and copper/zinc ratio) and carotid artery intima-media thickness (CIMT) in HD patients. Fifty chronic HD patients without known atherosclerotic disease and 48 age- and sex-matched healthy individuals were included in the study. The serum levels of trace elements (iron, zinc, manganese, copper, and magnesium) and heavy metals (cobalt, cadmium, and lead) were measured by Atomic Adsorption Spectrophotometer (UNICAM-929). CIMT was assessed by carotid artery ultrasonography. The serum levels of iron, zinc, and manganese were lower; levels of copper, magnesium, cobalt, cadmium, lead, and copper/zinc ratio were higher in HD patients compared to controls. CIMT in HD patients were higher than the control group (0.64 +/- 0.11 vs 0.42 +/- 0.05, p < 0.001). There was a significant negative correlation between CIMT and serum levels of zinc (r = 0.70, p < 0.01), iron (r = -0.71, p < 0.01), and manganese (r = -0.47, p < 0.01), while there was a significant positive correlation between CIMT and serum levels of copper (r = 0.63, p < 0.01), magnesium (r = 0.77, p < 0.01), cobalt (r = 0.63, p < 0.01), cadmium (r = 0.48, p < 0.01), lead (r = 0.38, p < 0.01), and copper/zinc ratio (r = 0.68, p < 0.01). A linear regression analysis showed that serum levels of magnesium, cadmium, lead, and copper/zinc ratio were still significantly and positively correlated with CIMT. We propose that copper/zinc ratio, magnesium and toxic metals cadmium and lead are independent determinants of CIMT in maintenance HD patients without known atherosclerotic disease. PMID- 21647753 TI - Altered uptake and biological half-lives of 65Zn on arsenic exposure--modulation by zinc treatment. AB - The present study revealed the effects of zinc on the biokinetics of (65)Zn in rats following arsenic intoxication. The animals were segregated into four groups: group I--untreated controls, group II--arsenic treated (100 ppm as NaAsO(2) in drinking water), group III--zinc treated (227 mg ZnSO(4) per liter drinking water), and group IV--arsenic + zinc treated. Each rat was injected intraperitoneally with 1.85 MBq radioactivity of (65)Zn following 3 months of different treatments, and the radioactivity was determined using a suitably shielded scintillation counter. Arsenic treatment showed a significant increase in the fast component (Tb(1)) of the biological half-life of (65)Zn in liver, which remained unaltered in the whole body. Furthermore, arsenic treatment decreased significantly the slow component (Tb(2)) in the whole body, which remained unchanged in the liver. However, zinc supplementation to arsenic-treated rats normalized Tb(1) in the liver, but caused no change in Tb(2) in the whole body. Furthermore, the uptake values of (65)Zn were significantly increased in the liver, brain, kidney, and intestine following arsenic treatment, and the values in the liver and brain were decreased by zinc. Hence, zinc plays a significant role in regulating the biokinetics of (65)Zn in the liver and the whole body of arsenic-intoxicated rats. PMID- 21647754 TI - Micromineral source affects intestinal morphology but not feather formation in pigeons. AB - During 7 weeks, two groups of eight pigeons received Zn, Cu, Mn and Se either as inorganic salts or organically bound to investigate the effect of micromineral source on two fast processes in avian metabolism: feather regrowth and intestinal renewal. Increased plasma Zn with reduced hepatic Cu in the organically bound minerals group suggests improved Zn bio-availability with a net antagonistic effect on Cu. Micromineral source did not affect feather regrowth, organ weight or intestinal pH, but the increased villus height with similar crypt depth pointed to increased absorptive surface. PMID- 21647755 TI - Lead (Pb)-induced regulation of growth, photosynthesis, and mineral nutrition in maize (Zea mays L.) plants at early growth stages. AB - The phytotoxic effects of lead (Pb) on seed germinability, seedling growth, photosynthetic performance, and nutrient accumulation (K(+) and Cu(2+)) in two maize genotypes (EV-1098 and EV-77) treated with varying levels of PbSO(4) (0.01, 0.1, and 1.0 mg L(-1)) were appraised in this study. In the seed germination experiment, lead stress significantly reduced seed germination percentage and index, plumule and radicle lengths as well as fresh and dry weights in both genotypes. In the second experiment, lengths and fresh and dry weights of shoots and roots decreased due to Pb in both genotypes with increase in plant age. Higher Pb levels also decreased photosynthetic rate (A), water use efficiency (A/E), and intrinsic water use efficiency (A/g(s)), but increased transpiration rate (E) and C(i)/C(a) ratio as a result of increase in stomatal conductance (g(s)). The concentrations of K(+) and Cu(2+) decreased in root, stem, and leaves of both genotypes, which could be a direct consequence of multifold increase in Pb concentration in these tissues. Overall, cv. EV-1098 had better Pb tolerance potential than EV-77 because the former genotype showed less reduction in seed germinability parameters, photosynthetic performance, and K(+) and Cu(2+) accumulation in shoot and root under lead stress. PMID- 21647756 TI - Effects of aluminum exposure on serum sex hormones and androgen receptor expression in male rats. AB - The effects of aluminum (Al) exposure on reproductive functions of male rats were investigated. Forty male Wistar rats (4 weeks old) weighing 75-95 g were randomly divided into four groups and orally exposed to 0 (control group GC), 64.18 (low dose group GL), 128.36 (middle-dose group GM), and 256.72 (high-dose group GH) mg/kg aluminum trichloride in drinking water for 120 days. The levels of testosterone (T), follicle-stimulating hormone (FSH), and luteinizing hormone (LH) were determined by radioimmunoassay. The androgen receptor (AR) expressions in testes were detected respectively by immunohistochemistry and time quantitative PCR. Results showed that the levels of T and LH in GM and GH were lower than those in GC (P < 0.05), but there were no significant changes in FSH level in all Al-treated groups (P > 0.05). AR protein expressions in GM and GH were lower than those in GC (P < 0.05), and there was a dose-response relationship between Al-exposure doses and AR protein expressions. The levels of AR mRNA expressions were lower in all Al-treated groups than those of GC (P < 0.05). The results indicate that Al can cause endocrinal disorders and interfere with AR expression, which suppresses development and functional maintenance of the testes. PMID- 21647757 TI - Determination of trace element level in different tissues of the leaping mullet (Liza saliens, Mugilidae) collected from Caspian Sea. AB - The concentrations of Cr, Cu, Fe, Mn, Ni, Pb, Cd, and Zn were determined in the brain, heart, liver, gill, gonad, spleen, kidney, and red and white muscles of Liza saliens (leaping mullet). Trace element levels in fish samples were analyzed by flame atomic absorption spectrometry. Among the non-essential metals, the levels of Ni and Pb in the tissues were higher than limits for fish proposed by FAO/WHO, EU, and TFC. Generally, the levels of the non-essential metals were much higher than those of manganese in the red and white muscles. Fe distribution pattern in tissues was in order of spleen > liver > heart > gill > brain > kidney > gonad > red muscle > white muscle. Red muscle was not within the safe limits for human consumption because non-essential metal (Ni, Pb) contents were higher than standard limits. PMID- 21647758 TI - Intervention of selenium on chronic fluorosis-induced injury of blood antioxidant capacity in rats. AB - This study was conducted to further explore the effects of selenium on the blood antioxidant capacity in rats exposed to fluoride to find out the optimal dosage level of selenium. Animals were divided into prevention sequence (Selenium -> NaF, water -> NaF) and treatment sequence (NaF -> Selenium, NaF -> water) (sodium fluoride 50 mg/L; sodium selenite 0.375, 0.75, 1.5 mg/L). The exposure time was 12 months. Then, the fluidity of erythrocyte membrane by electron spin resonance was analyzed, and the blood was collected for GSH-Px and SOD activity, total antioxidant capacity (T-AOC) and uric acid assay, sialic acid and MDA content. The results showed that, compared with control group, GSH-Px activity and T-AOC level increased significantly (P < 0.05), and SOD activity was raised in varying degrees in prevention and treatment groups, respectively. Uric acid level was up regulated, but no significant differences were observed (P > 0.05). The fluidity of erythrocyte membrane showed significant increase (P < 0.05). As evident in this study, when the dose of selenium was 0.75 mg/L, all the activities of antioxidant enzymes increased significantly in prevention sequence; but in treatment sequence, the optimum intervention concentration was 1.5 mg/L. On the basis of results, the preventive effect of selenium was superior to treatment effect on the oxidative stress induced by an overdose of fluoride. PMID- 21647759 TI - Aortic intimal fold in Takayasu arteritis causing obstruction of left coronary ostium. AB - An 11-year-old girl with Takayasu arteritis was presented with recurrent chest pain. A transthoracic echocardiogram showed moderate to severe aortic regurgitation with impaired cardiac function. Cardiac catheterization revealed the total occlusion of the small right coronary artery and left main stenosis of 40%. An intraoperative transesophageal echocardiogram showed a redundant intimal fold partially covering the left coronary ostium and this was further confirmed from the intraoperative finding. This is a case report of recurrent angina associated with obstruction of left main coronary artery orifice by an intimal fold secondary to Takayasu arteritis. PMID- 21647760 TI - Insertion-deletion polymorphisms--utilization on forensic analysis. AB - Insertion-deletion (INDEL) markers are very frequent in the human genome and present several advantages for population and forensic studies, such as low mutation rates, easy interpretation, small amplicons, easy genotyping, and the possibility of using multiplex PCR. The great adaptability of INDELs for amplification of low copy number or degraded DNA allows its using as an interesting platform of genetic identity by DNA in forensic cases. In the present study, we tested the ability of 48 diallelic INDEL markers on genotyping forensic samples collected from different biological samples related to criminal cases. Moreover, we evaluated the lowest DNA concentration with which there was amplification of all markers from each one of three indel-plex panels. When comparing the performances obtained by the indel-plex panels described in this study with results obtained using Identifiler(r) kit (Applied Biosystems) related to forensic samples, as well as to control samples with different concentrations of DNA, we observed superior efficiency on samples with low copy number or in the presence of inhibitors. PMID- 21647761 TI - Factors predictive of the status of sentinel lymph nodes in melanoma patients from a large multicenter database. AB - BACKGROUND: Numerous predictive factors for cutaneous melanoma metastases to sentinel lymph nodes have been identified; however, few have been found to be reproducibly significant. This study investigated the significance of factors for predicting regional nodal disease in cutaneous melanoma using a large multicenter database. METHODS: Seventeen institutions submitted retrospective and prospective data on 3463 patients undergoing sentinel lymph node (SLN) biopsy for primary melanoma. Multiple demographic and tumor factors were analyzed for correlation with a positive SLN. Univariate and multivariate statistical analyses were performed. RESULTS: Of 3445 analyzable patients, 561 (16.3%) had a positive SLN biopsy. In multivariate analysis of 1526 patients with complete records for 10 variables, increasing Breslow thickness, lymphovascular invasion, ulceration, younger age, the absence of regression, and tumor location on the trunk were statistically significant predictors of a positive SLN. CONCLUSIONS: These results confirm the predictive significance of the well-established variables of Breslow thickness, ulceration, age, and location, as well as consistently reported but less well-established variables such as lymphovascular invasion. In addition, the presence of regression was associated with a lower likelihood of a positive SLN. Consideration of multiple tumor parameters should influence the decision for SLN biopsy and the estimation of nodal metastatic disease risk. PMID- 21647762 TI - Oncologic outcomes and survival in pT0 tumors after radical cystectomy in patients without neoadjuvant chemotherapy: results from a large multicentre collaborative study. AB - PURPOSE: To assess the postsurgical survival of patients with urothelial carcinoma of the bladder with pT0 tumor at pathologic examination of cystectomy specimens. METHODS: A multi-institutional, retrospective database was analyzed with data from 4758 radical cystectomy (RC) patients who underwent RC without neoadjuvant chemotherapy and who were diagnosed with pT0 on the basis of the pathologic specimen. Survival curves were estimated. A multivariate Cox model was used to evaluate the association between prognosis factors and disease recurrence or survival. RESULTS: Overall, 258 patients (5.4%) were included in the study. The median age was 64 years. At last resection, 171 tumors were invasive (at least pT2), and 87 were not. Median follow-up was 51 months. At multivariate analysis, initial location of the tumor and absence of lymphadenectomy were associated with tumor recurrence (P = 0.03 and P = 0.005, respectively) and specific mortality (P = 0.005 and 0.001, respectively). The main limitation of the study is its retrospective design, which is due to the rarity of this situation. Cancer-specific and recurrence-free survival rates were 89 and 85%, respectively, at 5 years and 82 and 80%, respectively, at 10 years. CONCLUSIONS: Despite acceptable oncological outcomes, patients with a pT0 tumor at the time of RC are still at risk of recurrence and progression and should not be considered to be entirely cured. In this population, stringent follow-up according to current recommendations should be effective. PMID- 21647763 TI - A 10-year trend analysis of sentinel lymph node frozen section and completion axillary dissection for breast cancer: are these procedures becoming obsolete? AB - BACKGROUND: Recent results from the ACOSOG Z0011 trial question the use of intraoperative frozen section (FS) during sentinel lymph node (SLN) biopsy and the role of axillary dissection (ALND) for SLN-positive breast cancer patients. Here we present a 10-year trend analysis of SLN-FS and ALND in our practice. METHODS: We reviewed our prospective SLN database over 10 years (1997-2006, 7509 SLN procedures) for time trends and variation between surgeons in the use of SLN FS and ALND in patients with cN0 invasive breast cancer. RESULTS: Use of SLN-FS decreased from 100% to 62% (P < 0.0001) and varied widely by surgeon (66% to 95%). There were no statistically significant trends in the performance of ALND for patients with SLN metastases detected by FS (n = 1370, 99-99%) or routine hematoxylin and eosin (H&E) (n = 333; 69-77%), but only for those detected by serial section H&E with or without immunohistochemistry (n = 438; 73-48%; P = 0.0054) or immunohistochemistry only (n = 294; 48-28%; P < 0.0001). These trends coincided with an increase in the proportion of completion versus immediate ALND (30-40%; P = 0.0710). CONCLUSIONS: Over 10 years, we have observed a diminishing rate of SLN-FS and, for patients with low-volume SLN metastases, fewer ALND, trends that suggest a more nuanced approach to axillary management. If the Z0011 selection criteria had been applied to our cohort, 66% of SLN-FS (4159 of 6327) and 48% of ALND (939 of 1953) would have been avoided, sparing 13% of all patients the morbidity of ALND. PMID- 21647764 TI - Stroke prevention-surgical and interventional approaches to carotid stenosis. AB - Extracranial carotidartery stenosis is an important cause of stroke that often needs treatment with carotid revascularization. To prevent stroke recurrence, carotid endarterectomy has been well-established for many years in treating symptomatic high- and moderate-grade stenosis. Carotid stenting is an appealing, less invasive alternative to carotid endarterectomy, and several recent trials have compared the efficacy of the 2 procedures in patients with carotid stenosis. Carotid artery stenting has emerged as an important mode of therapy for high-risk patients with symtomatic high-grade stenosis. This review focuses on the current data available that will enable the clinician to decide optimal treatment strategies for patients with carotid stenosis. PMID- 21647765 TI - Ginsenoside rd in experimental stroke: superior neuroprotective efficacy with a wide therapeutic window. AB - Ginsenoside Rd (Rd), one of the main active ingredients in Panax ginseng, has been demonstrated to protect against ischemic cerebral damage in vitro and in vivo. In this study, we aimed to further define the preclinical characteristics of Rd. We show that Rd passes the intact blood-brain barrier and exerts protection in both transient and permanent middle cerebral artery occlusion (MCAO) in rats. In the dose-response study, Rd (10-50 mg/Kg) significantly reduced the infarct volume on postoperative days (PODs) 1, 3, and 7. This protection was associated with an improved neurological outcome for as many as 6 weeks after transient MCAO, as assessed by modified neurological severity score, modified sticky-tape test, and corner test. For comparison, Rd was significantly more effective than edaravone and slightly more effective than N-tert-butyl-alpha phenylnitrone (PBN). In the therapeutic window study, Rd exhibited remarkable neuroprotection, even when administered for as many as 4 h after the recirculation of transient MCAO or after the onset of permanent MCAO. Furthermore, in female rats or 16-month-old male rats, the salutary effects of Rd were also observed. These findings suggest Rd is a promising neuroprotectant and provide support for future clinical studies to confirm whether Rd is beneficial in ischemic stroke. PMID- 21647766 TI - Epidural analgesia in open resection of colorectal cancer: is there a clinical benefit? a retrospective study on 1,470 patients. AB - BACKGROUND: Epidural analgesia (EA) is effective for postoperative pain relief and results in an earlier recovery from postoperative paralytic ileus. This study evaluated the influence of epidural analgesia on the postoperative 30-day mortality and morbidity after open colorectal cancer resection. METHODS: A retrospective observational study was performed at a single, tertiary hospital. All patients with an open colorectal cancer surgery between 1991 and 2008 were identified from the hospital database. RESULTS: Of the 1,470 patients included in the study, 838 (57.0%) received an EA. Mortality was lower after EA (1.5% vs. 5.7%, p < 0.001). Risk of pneumonia was reduced after EA (odds ratio (OR), 0.45; 95% confidence interval (CI), 0.28-0.74; p = 0.001), but not the risk of anastomotic leakage (OR, 1.18; 95% CI, 0.76-1.81; p = 0.465) or surgical site infections (OR, 1.09; 95% CI, 0.74-1.60; p = 0.663). A subgroup analysis of 427 patients operated on after 2002 (reflecting improved perioperative management) yielded similar results. However, no significant reduction in mortality was observed in the subgroup analysis. CONCLUSION: For patients with open colorectal cancer surgery, the application of EA leads to a reduction in pneumonia. Although this is only a retrospective study, it strongly supports the use of EA. PMID- 21647767 TI - Peptide absorption after massive proximal small bowel resection: mechanisms of ileal adaptation. AB - BACKGROUND: Protein absorption occurs as di- and tri-peptides via H(+)/peptide co transporter-1 (PepT1). AIM: The aim of this study is to identify mechanisms of ileal adaptation after massive proximal enterectomy. HYPOTHESIS: Ileal adaptation in uptake of peptides is mediated through upregulation of PepT1 gene expression. STUDY DESIGN: Rats underwent 70% jejunoileal resection. Total mucosal cellular levels of messenger RNA (mRNA) and protein and transporter-mediated uptake per centimeter of the di-peptide glycyl-sarcosine (Gly-Sar) were compared in remnant ileum 1 and 4 weeks postoperatively to control and to 1-week sham laparotomy rats. Histomorphology, food consumption, and weights of rats were monitored. RESULTS: After 70% resection, although mRNA per cell for PepT1 decreased at 1 week (p = 0.002), expression of mRNA at 4 weeks and protein at 1 and 4 weeks in remnant ileum were unchanged (p > 0.1). Ileal Gly-Sar uptake (V (max)-nanomoles per centimeter per minute, i.e., number of transporters per centimeter) increased at 1 and 4 weeks compared to control and 1-week sham (p < 0.05 each); K (m) (i.e., transporter function) was unchanged. Villous heights (millimeters) in remnant ileum increased at 1- and 4-week time points over controls (0.45 and 0.57 vs 0.21, resp; p < 0.001). CONCLUSIONS: Ileal adaptation to proximal resection for peptide absorption occurs through cellular proliferation (hyperplasia) and not through cellular upregulation of PepT1 mRNA or protein per enterocyte. PMID- 21647768 TI - TEM as a Platform for NOTES. PMID- 21647769 TI - Indications and techniques of transanal endoscopic microsurgery (TEMS). AB - Transanal endoscopic microsurgery (TEMS) has recently reemerged as a valuable technique for the management of rectal neoplasms - both benign and malignant. Since the original description of this technique in the early 1980s, TEMS has emerged as the approach of choice for most benign rectal tumors because of the excellent views provided and superior dissection techniques possible when compared to traditional transanal excision. Many published reports demonstrate that the lowest rates of recurrence are associated with TEMS probably because of full-thickness excision with negative margins. Increasingly, TEMS is being applied to primary rectal cancer when used alone as a full-thickness excision alone or in combination with additional therapies, depending on tumor stage. There is now a significant evidence base to suggest that this approach should be considered as part of a multidisciplinary approach to rectal cancer. This paper describes indications and techniques for this technology. PMID- 21647770 TI - Primary colorectal lymphoma-clinical outcomes in a population-based series. AB - PURPOSE: The purpose of this study was to investigate the characteristics and the outcomes of primary colorectal lymphomas using a population-based registry. METHODS: All cases of colorectal lymphoma diagnosed between 1980 and 2007 were identified using a provincial cancer registry. Patients meeting Dawson's criteria and having a negative bone marrow biopsy were included. RESULTS: One hundred ten cases of colorectal lymphoma were identified, 43 met the inclusion criteria. The majority of patients was male (86%), and the median age at diagnosis was 62 (range 26-82) years. Tumors were mostly located in the cecum (51.1%) and rectum (20.9%). The 5-year overall survival rate calculated by the Kaplan-Meier method was 57%. Age under 60 was associated with a better median survival time (265 vs 54 months; p < 0.0001). The surgical treatment was associated with a better overall survival compared to medical treatment alone (110 vs 56 months; p = 0.083). Tumors located in the rectum were associated with a decreased overall survival (41 months vs 110 months; p = 0.065). CONCLUSIONS: Primary colorectal lymphoma is a rare disease. The age at diagnosis is an important predictor of outcome. Surgical resection may be associated with improved survival. Rectal lymphoma appears to be associated with a worse outcome and may warrant more aggressive therapy. PMID- 21647771 TI - [Autoinflammatory diseases as cause of wound healing defects]. AB - Ulcerations of the skin and mucosal membranes are a common feature of autoinflammatory diseases. They can give raise to chronic wound healing defects and should be considered in the differential diagnosis of chronic skin ulcers. The increased activation of the innate immune system in the absence of an apparent provocation for inflammation is a hallmark of autoinflammatory diseases. Mutations and alterations of signaling pathways regulating the innate immune response to physical trauma/tissue damage result into an unrestrained activation of the inflammasome, which leads to increased activation of Interleukin-1. Uncontrolled recruitment and activation of myeloid effector cells within the wound site lead to the release of potent proteases that cause the degradation of structural components of the skin. The majority of these diseases respond well to immunosuppressive and immunomodulatory treatment regimes. Therapeutic resistance converts the acute inflammatory response into a chronic and non-resolving inflammatory process that leads to tissue degeneration. In this article we will focus on the review of those autoinflammatory diseases that often display ulcerative cutaneous and aphthous lesions including pyoderma gangrenosum, Behcet disease, PAPA syndrome and hyperimmunoglobulinemia D with periodic fever syndrome (HIDS). Furthermore, the article will be complemented by an overview of those inflammatory diseases that are associated with non-ulcerative cutaneous manifestations. PMID- 21647772 TI - Field observations of fish species susceptible to epizootic ulcerative syndrome in the Zambezi River basin in Sesheke District of Zambia. AB - A field investigation was conducted in the Sesheke District of Zambia along the Zambezi River to determine the fish species susceptible to epizootic ulcerative syndrome (EUS), a newly confirmed disease in Southern Africa. A total of 2,132 fishes were inspected for gross EUS-like lesions, of which 188 (8.82%; 95% CI=7.67-10.1%) were found with typical characteristic lesions of EUS. Of these 188 samples, 156 were found to have mycotic granulomas on histopathological analysis, representing 83.0% (95% CI=76.7-87.9%) of the initially identified in the laboratory through gross examination. The following 16 species of fish were examined and found with EUS lesions; Clarias ngamensis, Clarias gariepinus, Barbus poechii, Tilapia sparrmanii, Serranochromis angusticeps, Brycinus lateralis, Micralestes acutidens, Sargochromis carlottae, Hydrocynus vittatus, Phryngochromis acuticeps, Schilbe intermedius, Hepsetus odoe, Labeo lunatus, Oreochromis andersonii, Barbus unitaeniatus, and Barbus paludinosus. T. sparrmanii did not show any lesions, while the Clarias species were found to be the most afflicted with EUS. These results could be useful to fish farmers and organizations interested in improving aquaculture in the area. PMID- 21647773 TI - Seroprevalence and risk factors for Leptospirosis in goats in Uberlandia, Minas Gerais, Brazil. AB - The prevalence of Leptospirosis in goat herds of the State of Minas Gerais has seldom been studied. The present research had as its objectives: (1) investigate the seroprevalence of Leptospirosis in the county of Uberlandia, MG, (2) verify the Leptospirosis serovars, and (3) identify the risk factors associated with infection on the farms examined. Serum samples from 230 animals in 11 properties were tested using the microscopic agglutination test. An epidemiological examination furnished data for analysis regarding the risk factors. The prevalence of Leptospirosis was found to be 31.3% with variation from 1:100 to 1:800. The most frequent serovars were: Autumnalis (30.30%), Tarassovi (19.20%), Pyrogenes (13.13%), and Icterohaemorrhagiae (11.11%). The ages and races of the animals were among the risk factors found to be significantly correlated (P<0.05) with infection. At the farm level, the intensity of production, use of salaried workers, and association of other animals were all found to be related with the frequency of Leptospirosis. The results demonstrated that inadequate management was a factor which favored the occurrence of infection in the region of the study. PMID- 21647774 TI - Molecular typing of Brucella species isolates from livestock and human. AB - Although host specificity has been observed in different species of Brucella, crossing the animal host boundary is likely to occur at any time. In this study, Bruce ladder PCR and abortus-melitensis-ovis-suis (AMOS) PCR assays were used to characterize 47 Brucella isolates from Indian origin in order to know exact species for understanding epidemiology of brucellosis. Out of them, 28, 14, and 5 isolates were found to be Brucella abortus, Brucella melitensis, and Brucella suis, respectively. Further analysis by AMOS PCR has identified that all the B. abortus isolates belong to any one of the biovar 1, 2, or 4; of the five B. suis isolates, three belong to biovar 1 and two belong to any one of the biovar 2, 3, 4, or 5. Although this multiplex Bruce ladder PCR is useful in differentiating all Brucella species, elaborate study is required to further characterize the isolates at exact biovar level. PMID- 21647775 TI - Redox biochemistry of mammalian metallothioneins. AB - Metallothionein (MT) is a generic name for certain families of structurally rather variable metal-binding proteins. While purely chemical or biological approaches failed to establish a single physiologic function for MTs in any species, a combination of chemical and biological approaches and recent progress in defining the low but significant concentrations of cytosolic free zinc(II) ions have demonstrated that mammalian MTs function in cellular zinc metabolism in specific ways that differ from conventional knowledge about any other metalloprotein. Their thiolate coordination environments make MTs redox-active zinc proteins that exist in different molecular states depending on the availability of cellular zinc and the redox poise. The zinc affinities of MTs cover a range of physiologic zinc(II) ion concentrations and are modulated. Oxidative conditions make more zinc available, while reductive conditions make less zinc available. MTs move from the cytosol to cellular compartments, are secreted from cells, and are taken up by cells. They provide cellular zinc ions in a chemically available form and participate in cellular metal muffling: the combination of physiologic buffering in the steady state and the cellular redistribution and compartmentalization of transiently elevated zinc(II) ion concentrations in the pre-steady state. Cumulative evidence indicates that MTs primarily have a redox-dependent function in zinc metabolism, rather than a zinc dependent function in redox metabolism. PMID- 21647776 TI - Chemistry and biology of mammalian metallothioneins. AB - Metallothioneins (MTs) are a class of ubiquitously occurring low molecular mass, cysteine- and metal-rich proteins containing sulfur-based metal clusters formed with Zn(II), Cd(II), and Cu(I) ions. In mammals, four distinct MT isoforms designated MT-1 through MT-4 exist. The first discovered MT-1/MT-2 are widely expressed isoforms, whose biosynthesis is inducible by a wide range of stimuli, including metals, drugs, and inflammatory mediators. In contrast, MT-3 and MT-4 are noninducible proteins, with their expression primarily confined to the central nervous system and certain squamous epithelia, respectively. MT-1 through MT-3 have been reported to be secreted, suggesting that they may play different biological roles in the intracellular and extracellular space. Recent reports established that these isoforms play an important protective role in brain injury and metal-linked neurodegenerative diseases. In the postgenomic era, it is becoming increasingly clear that MTs fulfill multiple functions, including the involvement in zinc and copper homeostasis, protection against heavy metal toxicity, and oxidative damage. All mammalian MTs are monomeric proteins, containing two metal-thiolate clusters. In this review, after a brief summary of the historical milestones of the MT-1/MT-2 research, the recent advances in the structure, chemistry, and biological function of MT-3 and MT-4 are discussed. PMID- 21647777 TI - A cryo-crystallographic time course for peroxide reduction by rubrerythrin from Pyrococcus furiosus. AB - High-resolution crystal structures of Pyrococcus furiosus rubrerythrin (PfRbr) in the resting (all-ferrous) state and at time points following exposure of the crystals to hydrogen peroxide are reported. This approach was possible because of the relativity slow turnover of PfRbr at room temperature. To this end, we were able to perform time-dependent peroxide treatment of the fully reduced enzyme, under strictly anaerobic conditions, in the crystalline state. In this work we demonstrate, for the first time, that turnover of a thermophilic rubrerythrin results in approximately 2-A movement of one iron atom in the diiron site from a histidine to a carboxylate ligand. These results confirm that, despite the domain swapped architecture, the hyperthermophilic rubrerythrins also utilize the classic combination of iron sites together with redox-dependent iron toggling to selectively reduce hydrogen peroxide over dioxygen. In addition, we have identified previously unobserved intermediates in the reaction cycle and observed structural changes that may explain the enzyme precipitation observed for the all iron form of PfRbr upon oxidation to the all-ferric state. PMID- 21647778 TI - ISC-like [2Fe-2S] ferredoxin (FdxB) dimer from Pseudomonas putida JCM 20004: structural and electron-nuclear double resonance characterization. AB - The crystal structure of the ISC-like [2Fe-2S] ferredoxin (FdxB), probably involved in the de novo iron-sulfur cluster biosynthesis (ISC) system of Pseudomonas putida JCM 20004, was determined at 1.90-A resolution and displayed a novel tail-to-tail dimeric form. P. putida FdxB lacks the consensus free cysteine usually present near the cluster of ISC-like ferredoxins, indicating its primarily electron transfer role in the iron-sulfur cluster. Orientation selective electron-nuclear double resonance spectroscopic analysis of reduced FdxB in conjunction with the crystal structure has identified the innermost Fe2 site with a high positive spin population as the nonreducible iron retaining the Fe(3+) valence and the outermost Fe1 site as the reduced iron with a low negative spin density. The average g (max) direction is skewed, forming an angle of about 27.3 degrees (+/-4 degrees ) with the normal of the [2Fe-2S] plane, whereas the g (int) and g (min) directions are distributed in the cluster plane, presumably tilted by the same angle with respect to this plane. These results are related to those for other [2Fe-2S] proteins in different electron transport chains (e.g. adrenodoxin) and suggest a significant distortion of the electronic structure of the reduced [2Fe-2S] cluster under the influence of the protein environment around each iron site in general. PMID- 21647779 TI - Metabolic effects of pioglitazone in chemically-induced mammary carcinogenesis in rats. AB - In this paper, the effect of peroral antidiabetic pioglitazone, a thiazolidinedione derivate, on selected parameters of carbohydrate and lipid metabolism in N-methyl-N-nitrosourea-induced mammary carcinogenesis in female Sprague-Dawley rats was evaluated. Pioglitazone was administered in the diet at two concentrations (10 ppm and 100 ppm), the chemoprevention was initiated 12 days before carcinogenesis induction and lasted until the termination of the experiment. The experiment was terminated 17 weeks after carcinogenesis induction, selected organs and tissues were removed and weighed and basic metabolic and hormonal parameters were determined. Pioglitazone increased glycemia (without exceeding normal values) and glycogen concentration in both liver and heart muscle without altering insulinemia and increased triacylglycerol concentration in liver, these changes were more prominent in group with higher dose. Pioglitazone also reduced corticosterone serum concentration and attenuated lipid peroxidation. Pioglitazone and other glitazones may be useful in alleviation of unfavourable metabolic changes in cancer patients. PMID- 21647780 TI - Smoking and polymorphisms in xenobiotic metabolism and DNA repair genes are additive risk factors affecting bladder cancer in Northern Tunisia. AB - Cancer epidemiology has undergone marked development since the nineteen-fifties. One of the most spectacular and specific contributions was the demonstration of the massive effect of smoking and genetic polymorphisms on the occurrence of bladder cancer. The tobacco carcinogens are metabolized by various xenobiotic metabolizing enzymes, such as the super-families of N-acetyltransferases (NAT) and glutathione S-transferases (GST). DNA repair is essential to an individual's ability to respond to damage caused by tobacco carcinogens. Alterations in DNA repair genes may affect cancer risk by influencing individual susceptibility to this environmental exposure. Polymorphisms in NAT2, GST and DNA repair genes alter the ability of these enzymes to metabolize carcinogens or to repair alterations caused by this process. We have conducted a case-control study to assess the role of smoking, slow NAT2 variants, GSTM1 and GSTT1 null, and XPC, XPD, XPG nucleotide excision-repair (NER) genotypes in bladder cancer development in North Tunisia. Taken alone, each gene unless NAT2 did not appear to be a factor affecting bladder cancer susceptibility. For the NAT2 slow acetylator genotypes, the NAT2*5/*7 diplotype was found to have a 7-fold increased risk to develop bladder cancer (OR = 7.14; 95% CI: 1.30-51.41). However, in tobacco consumers, we have shown that Null GSTM1, Wild GSTT1, Slow NAT2, XPC (CC) and XPG (CC) are genetic risk factors for the disease. When combined together in susceptible individuals compared to protected individuals these risk factors give an elevated OR (OR = 61). So, we have shown a strong cumulative effect of tobacco and different combinations of studied genetic risk factors which lead to a great susceptibility to bladder cancer. PMID- 21647781 TI - Primary peritoneal serous papillary carcinoma: a clinical and pathological study. AB - Primary peritoneal serous papillary carcinoma (PPSPC) is a rare primary tumor of the peritoneum that found predominantly in elderly and post-menopausal women. The aim of our study is to review the clinical and pathologic information of 22 patients, and then try to summarize clinical behavior and pathological characteristics of PPSPC, in order to be better recognized of this entity in future. We retrospectively reviewed the data from 22 patients with PPSPC treated at our hospital from 1992 to 2008. All paraffin blocks were recut for periodic acid-Schiff diastase and immunohistochemical staining for CD15, cytokeratin7(CK7), cytokeratin20(CK20), S-100 protein, carcinoembryonic antigen (CEA), CA125, estrogen receptor(ER) and progesterone receptor(PR). The median age of the patients at the time of surgical staging was 56 years (range, 32-77 years). The most common presenting symptoms were abdominal distension (59.1%) and ascites (63.6%). Pretreatment CA125 levels were significant elevated in 90.5% patients. Optimal debulking was performed in 18 patients. All patients were consequently treated with platinum-based chemotherapy. Response to treatment is promising, and the median overall survival of all patients was 21.0 months (95% CI 16.9, 25.1 months). The positive rate of immunohistochemical staining was CD15 95.5%, CK7 90.9%, S-100 protein 68.2%, CA125 59.1%, CK20 31.8%, ER 31.8%, CEA 27.3% and PR 9.1%, respectively. Gynecologist should be aware of PPSPC when abdominal distension, gross ascites and a raised level of CA125 in women without ovarian enlargement. Immunohistochemical staining might be helpful as accessory criteria for the differential diagnosis among the PPSPC, peritoneal malignant mesothelioma (PMM), primary epithelial ovarian carcinoma (PEOC) and peritoneal carcinomatosis from the gastrointestinal tumors (SPCGT). Cytoreductive surgery combined with pre/postoperative platinum-based chemotherapy may be effective for PPSPC patients. PMID- 21647782 TI - LipidomeDB data calculation environment: online processing of direct-infusion mass spectral data for lipid profiles. AB - LipidomeDB Data Calculation Environment (DCE) is a web application to quantify complex lipids by processing data acquired after direct infusion of a lipid containing biological extract, to which a cocktail of internal standards has been added, into an electrospray source of a triple quadrupole mass spectrometer. LipidomeDB DCE is located on the public Internet at http://lipidome.bcf.ku.edu:9000/Lipidomics . LipidomeDB DCE supports targeted analyses; analyte information can be entered, or pre-formulated lists of typical plant or animal polar lipid analytes can be selected. LipidomeDB DCE performs isotopic deconvolution and quantification in comparison to internal standard spectral peaks. Multiple precursor or neutral loss spectra from up to 35 samples may be processed simultaneously with data input as Excel files and output as tables viewable on the web and exportable in Excel. The pre-formulated compound lists and web access, used with direct-infusion mass spectrometry, provide a simple approach to lipidomic analysis, particularly for new users. PMID- 21647783 TI - Increasing donor ecosystem productivity decreases terrestrial consumer reliance on a stream resource subsidy. AB - Because nutrient enrichment can increase ecosystem productivity, it may enhance resource flows to adjacent ecosystems as organisms cross ecosystem boundaries and subsidize predators in recipient ecosystems. Here, we quantified the biomass and abundance of aquatic emergence and terrestrial spiders in a reference and treatment stream that had been continuously enriched with nitrogen and phosphorus for 5 years. Because we previously showed that enrichment increased secondary production of stream consumers, we predicted that aquatic emergence flux would be higher in the treatment stream, subsequently increasing the biomass and abundance of terrestrial spiders. Those increases were predicted to be greatest for spiders specializing on aquatic emergence subsidies (e.g., Tetragnathidae). By adding a (15)N stable isotope tracer to both streams, we also quantified nitrogen flow from the stream into the riparian community. Emergence biomass, but not abundance, was higher in the treatment stream. The average body size of emerging adult insects and the relative dominance of Trichoptera adults were also greater in the treatment stream. However, spider biomass did not differ between streams. Spiders also exhibited substantially lower reliance on aquatic emergence nitrogen in the treatment stream. This reduced reliance likely resulted from shifts in the body size distributions and community composition of insect emergence that may have altered predator consumption efficiency in the treatment stream. Despite nutrient enrichment approximately doubling stream productivity and associated cross-ecosystem resource flows, the response of terrestrial predators depended more on the resource subsidy's characteristics that affected the predator's ability to capitalize on such increases. PMID- 21647784 TI - Differential diagnosis of pericardial effusion after stem cell transplantation in acute myeloic leukemia. AB - The pathology underlying a pericardial effusion in a 24-year-old patient, who had suffered from acute myeloic leukemia 5 years previously and undergone chemotherapy followed by whole body radiation prior to allogeneic stem cell transplantation, could be identified by the careful analysis of pericardial cytology and epicardial biopsy guided by flexible pericardioscopy. Molecular, histological, cytochemical and immunological examination of the effusion and the epicardial biopsy for a viral or bacterial infection despite known CMV reactivation, or an effusion induced by radiation or graft-versus-host reaction, could be ruled out as possible causes of pericardial tamponade. The infiltration of CD 117-positive cells in the biopsied cardiac tissue revealed recurrent acute myeloic leukemia now also affecting the heart and the pericardium. An intrapericardial instillation of 1000 mg triamcinolone acetate at day 1 and 50 mg/m(2) cisplatin at day 3 effectively prevented the recurrence of tamponade, but could not prevent a lethal outcome 3 weeks later. PMID- 21647785 TI - Cardiovascular disease in cancer patients... only the tip of the iceberg? PMID- 21647786 TI - Episodic memory updating: the role of context familiarity. AB - We previously demonstrated that spatial context is a powerful reminder that can trigger memory updating (Hupbach, Hardt, Gomez, & Nadel in Learning & Memory, 15, 574-579 2008). In the present study, we asked whether the familiarity of the spatial context modulates the role of spatial context as a reminder. Since context familiarity can be easily manipulated in children, we chose 5-year-olds as study participants. In two experiments, we demonstrated robust memory-updating effects in children. Spatial context triggered incorporation of new information into old memories only when the context was unfamiliar. In highly familiar spatial environments (children's homes), spatial context did not initiate memory updating. Other reminders (the experimenter and a reminder question) became highly effective in familiar contexts. These findings shed further light on the specific conditions that trigger memory updating and support the view that the mechanisms underlying it are similar in children and adults. PMID- 21647787 TI - Predictive value of brain perfusion SPECT for rTMS response in pharmacoresistant depression. AB - PURPOSE: The aim of this study was to determine the predictive value of whole brain voxel-based regional cerebral blood flow (rCBF) for repetitive transcranial magnetic stimulation (rTMS) response in patients with pharmacoresistant depression. METHODS: Thirty-three right-handed patients who met DSM-IV criteria for major depressive disorder (unipolar or bipolar depression) were included before rTMS. rTMS response was defined as at least 50% reduction in the baseline Beck Depression Inventory scores. The predictive value of (99m)Tc-ethyl cysteinate dimer (ECD) single photon emission computed tomography (SPECT) for rTMS response was studied before treatment by comparing rTMS responders to non responders at voxel level using Statistical Parametric Mapping (SPM) (p < 0.001, uncorrected). RESULTS: Of the patients, 18 (54.5%) were responders to rTMS and 15 were non-responders (45.5%). There were no statistically significant differences in demographic and clinical characteristics (p > 0.10). In comparison to responders, non-responders showed significant hypoperfusions (p < 0.001, uncorrected) in the left medial and bilateral superior frontal cortices (BA10), the left uncus/parahippocampal cortex (BA20/BA35) and the right thalamus. The area under the curve for the combination of SPECT clusters to predict rTMS response was 0.89 (p < 0.001). Sensitivity, specificity, positive predictive value and negative predictive value for the combination of clusters were: 94, 73, 81 and 92%, respectively. CONCLUSION: This study shows that, in pharmacoresistant depression, pretreatment rCBF of specific brain regions is a strong predictor for response to rTMS in patients with homogeneous demographic/clinical features. PMID- 21647788 TI - Evaluation of CCK2 receptor binding ligands: the inheritance of Thomas Behr. PMID- 21647789 TI - Dimeric cationic amphiphilic polyproline helices for mitochondrial targeting. AB - PURPOSE: Efficient delivery of therapeutic biopolymers across cell membranes remains a daunting challenge. The development of cell-penetrating peptides (CPPs) has been useful; however, many CPPs are found trapped within endosomes, limiting their use as delivery agents. We optimize a class of CPPs, cationic amphiphilic polyproline helices (CAPHs), for direct transport into cells with mitochondrial localization through dimerization. METHODS: The CAPH P11LRR used for this study has been found to enter cells by two distinct pathways: an endocytotic pathway was favored at low concentrations; internalization by direct transport was observed at higher concentrations. CAPH was dimerized to probe if direct transport within cells may be enhanced through increased association of CAPH with the membrane and through the association of individual peptides within the membrane. RESULTS: The dimerization of the CAPH was found to significantly increase cellular uptake over its monomeric counterpart, with a concomitant lowering of the concentration threshold favoring direct transport. Evidence for direct transport within cells and mitochondrial localization was observed. CONCLUSIONS: CAPH cellular uptake efficiency can be significantly enhanced through peptide dimerization while favoring cell entry via direct transport at low concentration with low cell toxicity. PMID- 21647790 TI - Mechanism-based pharmacokinetic-pharmacodynamic modeling of the dopamine D2 receptor occupancy of olanzapine in rats. AB - PURPOSE: A mechanism-based PK-PD model was developed to predict the time course of dopamine D(2) receptor occupancy (D(2)RO) in rat striatum following administration of olanzapine, an atypical antipsychotic drug. METHODS: A population approach was utilized to quantify both the pharmacokinetics and pharmacodynamics of olanzapine in rats using the exposure (plasma and brain concentration) and D(2)RO profile obtained experimentally at various doses (0.01 40 mg/kg) administered by different routes. A two-compartment pharmacokinetic model was used to describe the plasma pharmacokinetic profile. A hybrid physiology- and mechanism-based model was developed to characterize the D(2) receptor binding in the striatum and was fitted sequentially to the data. The parameters were estimated using nonlinear mixed-effects modeling . RESULTS: Plasma, brain concentration profiles and time course of D(2)RO were well described by the model; validity of the proposed model is supported by good agreement between estimated association and dissociation rate constants and in vitro values from literature. CONCLUSION: This model includes both receptor binding kinetics and pharmacokinetics as the basis for the prediction of the D(2)RO in rats. Moreover, this modeling framework can be applied to scale the in vitro and preclinical information to clinical receptor occupancy. PMID- 21647791 TI - The effect of liposome encapsulation on the pharmacokinetics of recombinant secretory leukocyte protease inhibitor (rSLPI) therapy after local delivery to a guinea pig asthma model. AB - PURPOSE: Inhaled recombinant Secretory Leukocyte Protease Inhibitor (rSLPI) has shown potential for treatment of inflammatory lung conditions. Rapid inactivation of rSLPI by cathepsin L (Cat L) and rapid clearance from the lungs have limited clinical efficacy. Encapsulation of rSLPI within 1,2-Dioleoyl-sn-Glycero-3 [Phospho-L-Serine]:Cholesterol liposomes (DOPS-rSLPI) protects rSLPI against Cat L inactivation in vitro. We aimed to determine the effect of liposomes on rSLPI pharmacokinetics and activity in vitro and after local delivery to the airways in vivo. METHODS: Transport of DOPS-rSLPI and free-rSLPI across a polarised air liquid epithelial monolayer was measured. An asthma guinea pig model was administered either DOPS-rSLPI liposomes or free-rSLPI by intratracheal instillation. RESULTS: Apparent permeability (P(app)) of free-rSLPI was significantly higher at 4.9 x 10-6 cm/s than for DOPS-rSLPI, P(app) of 2.05 x 10 7 cm/s, confirmed by in vivo studies. Plasma rSLPI concentrations were highest in free-rSLPI-treated animals compared with those treated with DOPS-rSLPI; there also appeared to be a trend for higher intracellular rSLPI content in animals dosed with DOPS-rSLPI compared to free-rSLPI. Eosinophil influx was recorded as a measure of inflammation. Pre-dosing with either free-rSLPI or DOPS-rSLPI prevented inflammatory response to antigen challenge to levels comparable to control animals. CONCLUSION: Encapsulation of rSLPI in DOPS:Chol liposomes improves stability, reduces clearance and increases residence time in the lungs after local delivery. PMID- 21647792 TI - Matching children on the autism spectrum to classrooms: a guide for parents and professionals. AB - Meeting the needs of a learner with an autism spectrum disorder requires specialized expertise. Assessing the extent to which a potential program or classroom meets a child's needs is a source of serious challenge for parents and professionals alike. Indeed, identifying, prioritizing and agreeing upon the child's needs are complex questions for which there are no clear and straightforward answers. The process of establishing a match between a student and a placement must explore several primary dimensions: child, setting, and instructor variables, treatment philosophy and strategies, assessment and evaluation, and family needs and involvement. Additionally, there is a great deal of complexity considering how to interpret, integrate and apply empirical research findings and prominent professional opinions to develop sound and practical solutions. Discussion and agreement about the importance of each of these factors and how they apply in a specific situation forms the foundation of an interactive dialogue between service providers and families to create a "best fit" between student and program. PMID- 21647793 TI - Asperger through the looking glass: an exploratory study of self-understanding in people with Asperger's syndrome. AB - Hobson (Autism and the development of mind. Lawrence Erlbaum, Hove, UK 1993) has proposed that the cognitive and linguistic disabilities that characterise autism result from abnormalities in inter-subjective engagement during infancy, which in turn results in impaired reflective self-awareness. The aim of the present study was to test Hobson's hypothesis by examining self-understanding in Asperger's syndrome (AS) using Damon and Hart's (Self-understanding in childhood and adolescence. Cambridge University Press, Cambridge, 1988) model of self-concept. Ten participants with Asperger's syndrome were compared with ten non AS controls using the Self-understanding Interview (Damon and Hart in Self-understanding in Childhood and Adolescence. Cambridge University Press, Cambridge, 1988). The study found that the Asperger's group demonstrated impairment in the "self-as object" and "self-as-subject" domains of the Self-understanding Interview, which supported Hobson's concept of an impaired capacity for self-awareness and self reflection in people with ASD. The results are discussed with reference to previous research regarding the development of self-understanding in people with ASD. PMID- 21647794 TI - Biomaterials advances in patches for congenital heart defect repair. AB - This article reviews current applications and novel candidate biomaterials for use as tissue-engineered scaffolds in pediatric cardiac tissue engineering. This overview of different types of biomaterials includes naturally derived and synthetic polymers and their biological, physical, and biomechanical properties for the use as a patch or baffle for surgical reconstruction of congenital heart defects. Applications and characteristics of composite biomaterials are highlighted with their respective feasibilities in use as cardiac scaffolds. Currently, a wide range of biomaterials has been introduced for cardiovascular reconstruction for complex congenital cardiac defects. However, there are still many remaining challenges for engineered tissue implantations. PMID- 21647795 TI - Multicentric GISCoR Study "intensive clinical follow-up versus surgical radicalization after complete endoscopic polypectomy of a malignant adenoma" (SEC GISCoR). AB - Colorectal cancer screening programs result in an early diagnosis of the disease. In 2007, 250 malignant polyps were identified in Lombardy, out of 1,329 screen detected colorectal carcinomas. The Italian Group for Colorectal Cancer (GISCoR) promoted the multicentric study "Endoscopic Follow-up versus Surgical Radicalization of Malignant Polyps after Complete Endoscopic Polypectomy" (SEC GISCoR). The protocol was a multicentric, prospective, observational, non randomized study. It included patients diagnosed a colorectal malignant adenoma, after complete endoscopic removal. From November 2005 to September 2009, three participating centers enrolled 120 patients with malignant polyps after "complete" endoscopic polypectomy; malignant polyps were classified as "low risk" or "high risk". The study had two arms: "Intensive follow-up" (42 patients: 32 with low-risk and 10 with high-risk polyps) and "Surgical radicalization" (78 patients: 5 with low-risk and 73 with high-risk polyps). Data were collected using an online CRF. Overall, 37/120 polyps (30.8%) were low risk and 83/120 (69.2%) were high risk. 42 out of 120 patients (35%) were enrolled in the "clinical follow-up" arm, while 78/120 (65%) entered the surgery arm. In 15 cases, patients were not enrolled in the correct arm, according to the criteria agreed upon before starting the study. There still is a high incidence (11.5%) of pathological mismatches. No clinical event was reported in 2.9 years of follow up. In conclusion, some differences emerged in the management of patients with malignant polyps among participating centers (p < 0.001), mismatches can be explained by high surgical risk or patient's choice. Only in 5 cases (4.2%), did data analysis not allow to exactly determine the reason for a choice different from protocol criteria. The availability of new risk factors and the evidence of pathological mismatches confirmed the need for future studies on this issue. PMID- 21647796 TI - Recurrent adenoid cystic carcinoma in the liver: a repeated laparoscopic surgical approach. AB - Adenoid cystic carcinoma (ACC) is characterized by a particularly aggressive behavior even many years after resection of primary tumor. The evolution of metastasis dramatically affects the final outcome but resection should always be evaluated. Herein is described a case of aggressive ACC of the parotid gland in a 30-year-old female. She developed local recurrence and lung metastases; then, she also developed two liver metastasis 112 and 132 months after the resection of the primitive cancer of the parotid gland. Both lesions were successfully managed by a laparoscopic approach. Intra-abdominal adhesions after the first surgery were mild, allowing an easier access for the second laparoscopic liver resection. At 1 year follow-up, the patient is liver disease free with a stable lung disease. To our knowledge, this is the first report of a double laparoscopic liver resection for parotid gland's ACC metachronous metastases. Patients with resected ACC need a strict and lifelong follow-up after the resection of the primitive cancer. Also for ACC, a laparoscopic approach to liver metastasis should always be considered as a viable alternative to open surgery. In our experience of over 90 cases, laparoscopic surgery causes less adhesions, allowing an easier approach for repeated resections. PMID- 21647797 TI - Prescription medication borrowing among adult patients at an urban medical center. AB - Prescription medication borrowing can result in adverse health outcomes. We aimed to study the patterns of borrowing prescription medications in an adult urban population seeking healthcare in the outpatient, emergency, and inpatient units of an urban medical center. Participants indicated whether they (1) had a primary care doctor, medical insurance, a prior history of substance abuse, psychiatric disorders, or chronic pain; and (2) had borrowed a prescription medication. If so, they noted the medication obtained, source, frequency of use, and reasons why they had not obtained a prescription from a licensed medical provider. Of the 641 participants, most were African American (75%), urban residents (75%), high school educated or less (71%), and lacked full-time employment (68%). Many had health insurance (90%) and had recently seen their primary medical provider (75%). Eighteen percent reported ever borrowing a prescription medication. On multivariate analysis, history of chronic pain was marginally associated with increased medication borrowing (odds ratio [OR] = 1.58) while having Medicare insurance (OR = 0.436) or a primary care medical provider routinely ask about medication usage (OR = 0.589) were significantly associated with decreased medication borrowing. The most commonly obtained medications were for pain (74%), usually in the form of opioids, and were obtained from a family member (49%) or friend (38%). Thirty-five percent of those who borrowed medications did so more than once a year, with lack of convenient access to medical care the most frequently cited reason for use (67%). Only a third of those who borrowed medications had informed their primary medical providers of the behavior. In conclusion, borrowing prescription medications is a common behavior in the population studied. Further research is warranted into interventions to reduce such use, especially the impact of methods to improve the convenience of contacting licensed medical providers. PMID- 21647798 TI - Housing quality, housing instability, and maternal mental health. AB - Poor housing conditions and residential instability have been associated with distress among women; however, this association could be the result of other social factors related to housing, such as intimate partner violence (IPV) and economic hardship. We examined associations of housing conditions and instability with maternal depression and generalized anxiety disorder (GAD) while accounting for IPV and economic hardship in the Fragile Families and Child Wellbeing Study (N = 2,104). In the third study wave, interviewers rated indoor housing quality, including housing deterioration (e.g., peeling paint and holes in floor) and housing disarray (e.g., dark, crowded, and noisy). Mothers reported whether they had moved more than twice in the past two years, an indicator of housing instability. A screening for depression and GAD was obtained from questions derived from the Composite International Diagnostic Interview-Short Form in the second and third study waves. IPV and economic hardship were assessed through questionnaire. In this sample, 16% of women were classified as having probable depression and 5% as having probable GAD. In adjusted analyses, mothers experiencing housing disarray (odds ratio [OR], 1.3 [95% confidence interval (CI), 1.0, 1.7]) and instability (OR, 1.4 [95% CI, 1.2, 2.3]) were more likely to screen positive for depression. In addition, those experiencing housing instability were more likely to screen positive for GAD (OR 1.9 [95% CI, 1.2, 3.0]) even after adjusting for other social factors. No associations were noted between housing deterioration and maternal mental health. Similar associations were noted when incident cases of probable depression and GAD were examined. Housing instability and disarray, but not deterioration, are associated with screening positive for depression and generalized anxiety among women regardless of other social stressors present in their lives. Housing could potentially present a point of intervention to prevent mental health consequences among mothers and possibly their children. PMID- 21647799 TI - Role of interleukin 28B rs12979860 C/T polymorphism on the histological outcome of chronic hepatitis C: relationship with gender and viral genotype. AB - BACKGROUND: This study aimed to determine whether the single-nucleotide polymorphism (rs12979860 C/T) of the interleukin 28B (IL-28B) gene, which is associated with hepatitis C virus (HCV) clearance, is also associated with fibrosis in chronic HCV infection. METHODS: An RFLP-PCR technique was used to genotype 629 HCV-positive patients (200 with cirrhosis) and 428 healthy control subjects. RESULTS: The genotype frequencies in the controls and chronic hepatitis C patients were as follows: C/C 47.0% vs. 32.6%, C/T 41.8% vs. 52.8% and T/T 11.2% vs. 14.6% (p < 0.0001). The C allele frequency was higher in HCV-2- (0.635) and 3- (0.692) infected patients in comparison to those infected with HCV-1 (0.550) or 4-5 (0.600) (p < 0.001). Infected T/T homozygotes had a mean staging score higher than other patients (3.50 vs. 3.04, p < 0.05). CONCLUSIONS: IL-28B rs12979860 C/T polymorphism is associated with a greater likelihood of HCV persistence, particularly in HCV genotypes 1 and 4. The T allele affects the severity of liver fibrosis. PMID- 21647800 TI - Investigation into stability of poly(vinyl alcohol)-based Opadry(r) II films. AB - Poly(vinyl alcohol) (PVA)-based formulations are used for pharmaceutical tablet coating with numerous advantages. Our objective is to study the stability of PVA based coating films in the presence of acidic additives, alkaline additives, and various common impurities typically found in tablet formulations. Opadry(r) II 85F was used as the model PVA-based coating formulation. The additives and impurities were incorporated into the polymer suspension prior to film casting. Control and test films were analyzed before and after exposure to 40 degrees C/75% relative humidity. Tests included film disintegration, size-exclusion chromatography, thermal analysis, and microscopy. Under stressed conditions, acidic additives (hydrochloric acid (HCl) and ammonium bisulfate (NH(4)HSO(4))) negatively impacted Opadry(r) II 85F film disintegration while NaOH, formaldehyde, and peroxide did not. Absence of PVA species from the disintegration media corresponded to an increase in crystallinity of PVA for reacted films containing HCl. Films with NH(4)HSO(4) exhibited slower rate of reactivity and less elevation in melting temperature with no clear change in melting enthalpy. Acidic additives posed greater risk of compromise in disintegration of PVA-based coatings than alkaline or common impurities. The mechanism of acid-induced reactivity due to the presence of acidic salts (HCl vs. NH(4)HSO(4)) may be different. PMID- 21647801 TI - Seizure and mental status change after surgery for pelvic organ prolapse. AB - The syndrome of inappropriate antidiuretic hormone secretion (SIADH) is a well documented postoperative complication but infrequently seen after gynecologic procedures. Clinical presentation is often insidious, and neurologic sequelae may be severe, especially in women. We report the first published case of postoperative hyponatremia due to SIADH after vaginal reconstructive surgery. Our patient sustained a tonic-clonic seizure on postoperative day 2 in the setting of profound hyponatremia. Early detection and prompt treatment allowed her to achieve a complete recovery. Familiarity with uncommon complications of vaginal surgery is important. Prompt initiation of treatment can prevent potentially fatal consequences. PMID- 21647802 TI - Sample preparation procedure for the determination of polycyclic aromatic hydrocarbons in petroleum vacuum residue and bitumen. AB - This paper describes a novel method of sample preparation for the determination of trace concentrations of polycyclic aromatic hydrocarbons (PAHs) in high boiling petroleum products. Limits of quantitation of the investigated PAHs in materials of this type range from tens of nanograms per kilogram to <20 MUg/kg. The studies revealed that in order to separate most of interferences from the analytes without a significant loss of PAHs, it is necessary to use size exclusion chromatography as the first step of sample preparation, followed by adsorption using normal-phase liquid chromatography. The use of orthogonal separation procedure described in the paper allows the isolation of only a group of unsubstituted and substituted aromatic hydrocarbons with a specific range of molar mass. The lower the required limit of quantitation of PAHs, the larger is the scale of preparative liquid chromatography in both steps of sample preparation needed. The use of internal standard allows quantitative results to be corrected for the degree of recovery of PAHs during the sample preparation step. Final determination can be carried out using HPLC-FLD, GC-MS, or HPLC-UV VIS/DAD. The last technique provides a degree of identification through the acquired UV-VIS spectra. PMID- 21647803 TI - Site-specific protein glycosylation analysis with glycan isomer differentiation. AB - Glycosylation is one of the most common yet diverse post-translational modifications. Information on glycan heterogeneity and glycosite occupancy is increasingly recognized as crucial to understanding glycoprotein structure and function. Yet, no approach currently exists with which to holistically consider both the proteomic and glycomic aspects of a system. Here, we developed a novel method of comprehensive glycosite profiling using nanoflow liquid chromatography/mass spectrometry (nano-LC/MS) that shows glycan isomer-specific differentiation on specific sites. Glycoproteins were digested by controlled non specific proteolysis in order to produce informative glycopeptides. High resolution, isomer-sensitive chromatographic separation of the glycopeptides was achieved using microfluidic chip-based capillaries packed with graphitized carbon. Integrated LC/MS/MS not only confirmed glycopeptide composition but also differentiated glycan and peptide isomers and yielded structural information on both the glycan and peptide moieties. Our analysis identified at least 13 distinct glycans (including isomers) corresponding to five compositions at the single N-glycosylation site on bovine ribonuclease B, 59 distinct glycans at five N-glycosylation sites on bovine lactoferrin, 13 distinct glycans at one N glycosylation site on four subclasses of human immunoglobulin G, and 20 distinct glycans at five O-glycosylation sites on bovine kappa-casein. Porous graphitized carbon provided effective separation of glycopeptide isomers. The integration of nano-LC with MS and MS/MS of non-specifically cleaved glycopeptides allows quantitative, isomer-sensitive, and site-specific glycoprotein analysis. PMID- 21647804 TI - Scrambled eyes? Disrupting scene structure impedes focal processing and increases bottom-up guidance. AB - Previous research has demonstrated that search and memory for items within natural scenes can be disrupted by "scrambling" the images. In the present study, we asked how disrupting the structure of a scene through scrambling might affect the control of eye fixations in either a search task (Experiment 1) or a memory task (Experiment 2). We found that the search decrement in scrambled scenes was associated with poorer guidance of the eyes to the target. Across both tasks, scrambling led to shorter fixations and longer saccades, and more distributed, less selective overt attention, perhaps corresponding to an ambient mode of processing. These results confirm that scene structure has widespread effects on the guidance of eye movements in scenes. Furthermore, the results demonstrate the trade-off between scene structure and visual saliency, with saliency having more of an effect on eye guidance in scrambled scenes. PMID- 21647805 TI - Transjugular intrahepatic porto-systemic shunt placement in a patient with left lateral split-liver transplant and mesenterico-left portal vein by pass placement. AB - This is a report of a successful placement of a transjugular intrahepatic porto systemic shunt in a young patient with previous left-lateral, split-liver transplant and mesenterico-left portal vein by pass placement after posttransplant extrahepatic portal vein thrombosis. PMID- 21647806 TI - Cytological sampling versus forceps biopsy during percutaneous transhepatic biliary drainage and analysis of factors predicting success. AB - PURPOSE: To assess the accuracy of cytological sampling and forceps biopsy in obstructing biliary lesions and to identify factors predictive of success. METHODS: Consecutive patients (n = 119) with suspected malignant inoperable obstructive jaundice treated with percutaneous transhepatic biliary drainage during 7 years were included (60 male; mean age 72.5 years). All patients underwent forceps biopsy plus cytological sampling by washing the forceps device in cytological solution. Patient history, procedural and pathological records, and clinical follow-up were reviewed. Statistical analysis included chi-square test and multivariate regression analysis. RESULTS: Histological diagnosis after forceps biopsy was more successful than cytology: Sensitivity was 78 versus 61%, and negative predictive value was 30 versus 19%. Cytology results were never positive when the forceps biopsy was negative. The cytological sample was negative and forceps sample positive in 2 cases of cholangiocarcinoma, 16 cases of pancreatic carcinoma, and 1 case of benign disease. Diagnostic accuracy was predicted by low bilirubin (p < 0.001), aspartate transaminase (p < 0.05), and white cell count (p <= 0.05). CONCLUSIONS: This technique is safe and effective and is recommended for histological diagnosis during PTBD in patients with inoperable malignant biliary strictures. Diagnostic yield is greater when bilirubin levels are low and there is no sepsis; histological diagnosis by way of forceps biopsy renders cytological sampling unnecessary. PMID- 21647808 TI - Percutaneous transhepatic removal of bile duct stones: results of 261 patients. AB - PURPOSE: To determine the effectiveness of percutaneous transhepatic removal of bile duct stones when the procedure of endoscopic therapy fails for reasons of anatomical anomalies or is rejected by the patient. METHODS: Between April 2001 and May 2010, 261 patients (138 male patients and 123 female patients; age range, 14-92 years; mean age, 64.6 years) with bile duct stones (common bile duct [CBD] stones=248 patients and hepatolithiasis=13 patients) were included in the study. First, percutaneous transhepatic cholangiography was performed and stones were identified. Percutaneous transhepatic balloon dilation of the papilla of Vater was performed. Then stones were pushed out into the duodenum with a Fogarty balloon catheter. If the stone diameter was larger than 15 mm, then basket lithotripsy was performed before balloon dilation. RESULTS: Overall success rate was 95.7%. The procedure was successful in 97.5% of patients with CBD stones and in 61.5% of patients with hepatolithiasis. A total of 18 (6.8%) major complications, including cholangitis (n=7), subcapsular biloma (n=4), subcapsular hematoma (n=1), subcapsular abscess (n=1), bile peritonitis (n=1), duodenal perforation (n=1), CBD perforation (n=1), gastroduodenal artery pseudoaneurysm (n=1), and right hepatic artery transection (n=1), were seen after the procedure. There was no mortality. CONCLUSION: Our experience suggests that percutaneous transhepatic stone expulsion into the duodenum through the papilla is an effective and safe approach in the nonoperative management of the bile duct stones. It is a feasible alternative to surgery when endoscopic extraction fails or is rejected by the patient. PMID- 21647807 TI - Single-session radiofrequency ablation of bilateral lung metastases. AB - PURPOSE: This retrospective study examined the feasibility and efficacy of bilateral lung radiofrequency ablation (RFA) performed in a single session. METHODS: From 2002-2009, patients with bilateral lung metastases were treated by RFA, where possible in a single session under general anesthesia with CT guidance. The second lung was punctured only if no complications occurred after treatment of the first lung. Five lung metastases maximum per patient were treated by RFA and prospectively followed. The primary endpoint was the evaluation of acute and delayed complications. Secondary endpoints were calculation of hospitalization duration, local efficacy, median survival, and median time to tumor progression. Local efficacy was evaluated on CT or positron emission tomography (PET) CT. RESULTS: Sixty-seven patients were treated for bilateral lung metastases with RFA (mean age, 62 years). Single-session treatment was not possible in 40 due to severe pneumothoraces (n = 24), bilateral pleural contact (n = 14), and operational exclusions (n = 2). Twenty-seven (41%) received single-session RFA of lesions in both lungs for 66 metastases overall. Fourteen unilateral and four bilateral pneumothoraces occurred (18 overall, 66.7%). Unilateral (n = 13) and bilateral (n = 2) chest tube drainage was required. Median hospitalization was 3 (range, 2-8) days. Median survival was 26 months (95% confidence interval (CI), 19-33). Four recurrences on RFA sites were observed (4 patients). Median time to tumor progression was 9.5 months (95% CI, 4.2-23.5). CONCLUSIONS: Although performing single-session bilateral lung RFA is not always possible due to pneumothoraces after RFA of first lung, when it is performed, this technique is safe and effective. PMID- 21647809 TI - Influence of CT-based attenuation correction in assessment of left and right ventricular functions with count-based gated blood-pool SPECT. AB - OBJECTIVE: Influence of CT-based attenuation correction (CT-AC) in assessment of left and right ventricular functions with count-based gated blood-pool SPECT (GBPS) was evaluated in a mixed population. METHODS: Thirty-two patients (81% male; mean age 56 +/- 12) referred for various symptoms or heart diseases were prospectively included. Data from 32 GBPS acquisitions were reconstructed using an iterative algorithm with (IRAC) and without (IRNC) CT-AC and analyzed using previously described segmentation software based on the watershed algorithm. LV and RV EF and volumes were assessed with and without CT-AC and compared. RESULTS: EF and volumes were correlated (P < .001 for all parameters with r = 0.97 for LV and RV EF; r = 0.96 for LV EDV; r = 0.98 for LV ESV; r = 0.96 for RV EDV and ESV). The mean values using IRAC and IRNC were different for all parameters with lower EF (respectively, 49% +/- 19% vs 51 %+/- 18%; P = .002 for LV EF and 50% +/ 14% vs 54%+/-15%; P < .001 for RV EF) and higher volumes (respectively, 142 +/- 41 mL vs 133 +/- 40 mL; P < .001 and 79 +/- 45 mL vs 71 +/- 42 mL; P < .001 for LV EDV and ESV; 91 +/- 32 mL vs 86 +/- 31 mL; P = .003 and 48 +/- 28 mL vs 43 +/- 26 mL; P < .001 for RV EDV and ESV). Limits of agreement were -11% to 6% and -11% to 4% for LV and RV EF. We found wider limits of agreement for LV volumes (-13 to 32 mL for EDV and -10 to 27 mL for ESV) than for RV volumes (-13 to 23 mL for EDV and -9 to 20 mL for ESV). Taking into account all volumes, we found a trend with a significant positive correlation between means and differences in volumes assessed with and without CT-AC. CONCLUSION: Assessment of both left and right ventricular functions by count-based GBPS with CT-AC showed higher volumes and lower EF. Differences were slight, especially for the range of normal to subnormal ventricular volumes. PMID- 21647810 TI - Source-constrained retrieval influences the encoding of new information. AB - Jacoby, Shimizu, Daniels, and Rhodes (Psychonomic Bulletin & Review, 12, 852-857, 2005) showed that new words presented as foils among a list of old words that had been deeply encoded were themselves subsequently better recognized than new words presented as foils among a list of old words that had been shallowly encoded. In Experiment 1, by substituting a deep-versus-shallow imagery manipulation for the levels-of-processing manipulation, we demonstrated that the effect is robust and that it generalizes, also occurring with a different type of encoding. In Experiment 2, we provided more direct evidence for context-related encoding during tests of deeply encoded words, showing enhanced priming for foils presented among deeply encoded targets when participants made the same deep encoding judgments on those items as had been made on the targets during study. In Experiment 3, we established that the findings from Experiment 2 are restricted to this specific deep judgment task and are not a general consequence of these foils being associated with deeply encoded items. These findings provide support for the source-constrained retrieval hypothesis of Jacoby, Shimizu, Daniels, and Rhodes: New information can be influenced by how surrounding items are encoded and retrieved, as long as the surrounding items recruit a coherent mode of processing. PMID- 21647811 TI - Evaluation of the mental health benefits of yoga in a secondary school: a preliminary randomized controlled trial. AB - The goal of this study was to evaluate potential mental health benefits of yoga for adolescents in secondary school. Students were randomly assigned to either regular physical education classes or to 11 weeks of yoga sessions based upon the Yoga Ed program over a single semester. Students completed baseline and end program self-report measures of mood, anxiety, perceived stress, resilience, and other mental health variables. Independent evaluation of individual outcome measures revealed that yoga participants showed statistically significant differences over time relative to controls on measures of anger control and fatigue/inertia. Most outcome measures exhibited a pattern of worsening in the control group over time, whereas changes in the yoga group over time were either minimal or showed slight improvements. These preliminary results suggest that implementation of yoga is acceptable and feasible in a secondary school setting and has the potential of playing a protective or preventive role in maintaining mental health. PMID- 21647812 TI - The implementation of smoking cessation counseling in substance abuse treatment. AB - Research on the implementation of smoking cessation counseling within substance abuse treatment organizations is limited. This study examines associations among counselors' implementation of therapy sessions dedicated to smoking cessation, organizational factors, and counselor-level variables. A two-level hierarchical linear model including organization- and counselor-level variables was estimated using survey data collected from 1,794 counselors working in 359 treatment organizations. Overall implementation of smoking cessation counseling was low. In the final model, implementation was positively associated with counselors' knowledge of the Public Health Service's clinical practice guideline, perceived managerial support, and belief that smoking cessation had a positive impact on recovery. Private versus public funding and presence of a formal smoking cessation program were organization-level variables which interacted with these counselor-level effects. These results highlight the importance of organizational contexts as well as counselors' knowledge and attitudes for effective implementation of smoking cessation counseling in substance abuse treatment organizations. PMID- 21647813 TI - Effect of training and instrument type on performance in single-incision laparoscopy: results of a randomized comparison using a surgical simulator. AB - PURPOSE: Single-incision laparoscopy (SIL) is potentially less invasive compared with standard laparoscopic surgery (LAP); however, it may be more technically challenging and have a longer learning curve. A two-phase study was conducted to examine the performance of standardized tasks on a surgical simulator by novices during a distributed training period. Phase 1 examined the effect of LAP-specific or SIL-specific training on skill acquisition for both techniques. Phase 2 compared the effectiveness and learning curves of additional instrument types for SIL (straight [STR] vs. dynamic articulating [D-ART]). METHODS: Medical students without previous surgical experience were randomized to LAP-specific training or SIL-specific training, using static articulating instruments [S-ART] for SIL. LAP and SIL scores on the peg transfer (PEG) and pattern cutting (CIRCLE) tasks from the Fundamentals of Laparoscopic Surgery (FLS) were measured at baseline and after four training sessions. In phase 2, a new group of subjects were randomized to SIL training using STR or D-ART instruments, with similar baseline and post training testing. FLS task scores were calculated and compared according to training regimen and instrument type. RESULTS: Forty-five subjects completed the study. All scores improved significantly during the training period. Improvement in LAP score was similar between LAP-trained and SIL-trained groups. Improvement of SIL score was better for the SIL-trained group. Final scores were better and the learning curve was shorter for LAP versus SIL technique, with no differences in SIL scores according to instrument type. CONCLUSIONS: LAP technique results in superior task performance with a shorter learning curve compared with SIL technique during a standardized training period. SIL-specific simulator training is better than LAP training alone to improve SIL performance. Neither S-ART nor D ART instruments for SIL are associated with improved performance or shorter learning curve compared with STR instruments. PMID- 21647814 TI - Risk factors for recurrence after transanal endoscopic microsurgery for rectal malignant neoplasm. AB - BACKGROUND: Indications and results of local excision of rectal lesions are currently under debate. Transanal endoscopic microsurgery (TEM), allowing a precise, full-thickness excision, could improve oncological results in early rectal tumors. METHODS: A prospective database was analyzed with the intent to identify risk factors for recurrence after TEM. RESULTS: Among 355 patients subjected to TEM, 107 had an adenocarcinoma: 48 pT1, 43 pT2, and 16 pT3. Comparing pre- and postoperative data, histological discrepancy was 20% and staging discrepancy was 34%. Mortality was nil, morbidity was 9%. Mean follow-up was 54.2 months (range = 12-164), follow-up rate was 100%. The 5-year disease free survival rate was 85.9, 78.4, and 49.4% for pT1, pT2, and pT3, respectively (p = 0.006). Recurrence rate was 0% (0/26) in pT1sm1 cancers and 22.7% (5/22) in sm2-3 (p < 0.05). A submucosal infiltration represented a significant risk factor for recurrences: 0% sm1, 16.7% sm2, and 30% sm3. Recurrence in pT2 was 0% in patients who had neoadjuvant therapy and 26% in the others. At univariate analysis, diameter, sm stage, pT stage, tumor grading, margin infiltration, and lymphovascular invasion demonstrated statistical significance. Multivariate analysis indicated sm stage, pT stage, and tumor grading as independent predictors of recurrence. CONCLUSIONS: TEM represents an effective curative treatment for pT1 sm1 rectal malignancies. pT1 sm2-3 patients should be considered high-risk cases if treated only by TEM. A consistent improvement in the preoperative assessment of the risk factors identified by the present study will be a crucial development for optimal treatment of early rectal cancers. PMID- 21647815 TI - The aldehyde dehydrogenase enzyme 7A1 is functionally involved in prostate cancer bone metastasis. AB - High aldehyde dehydrogenase (ALDH) activity can be used to identify tumor initiating and metastasis-initiating cells in various human carcinomas, including prostate cancer. To date, the functional importance of ALDH enzymes in prostate carcinogenesis, progression and metastasis has remained elusive. Previously we identified strong expression of ALDH7A1 in human prostate cancer cell lines, primary tumors and matched bone metastases. In this study, we evaluated whether ALDH7A1 is required for the acquisition of a metastatic stem/progenitor cell phenotype in human prostate cancer. Knockdown of ALDH7A1 expression resulted in a decrease of the alpha2(hi)/alphav(hi)/CD44(+) stem/progenitor cell subpopulation in the human prostate cancer cell line PC-3M-Pro4. In addition, ALDH7A1 knockdown significantly inhibited the clonogenic and migratory ability of human prostate cancer cells in vitro. Furthermore, a number of genes/factors involved in migration, invasion and metastasis were affected including transcription factors (snail, snail2, and twist) and osteopontin, an ECM molecule involved in metastasis. Knockdown of ALDH7A1 resulted in decreased intra-bone growth and inhibited experimentally induced (bone) metastasis, while intra-prostatic growth was not affected. In line with these observations, evidence is presented that TGF beta, a key player in cancer invasiveness and bone metastasis, strongly induced ALDH activity while BMP7 (an antagonist of TGF-beta signaling) down-regulated ALDH activity. Our findings show, for the first time, that the ALDH7A1 enzyme is functionally involved in the formation of bone metastases and that the effect appeared dependent on the microenvironment, i.e., bone versus prostate. PMID- 21647816 TI - The relationship between work-related stress and boundary-related stress within the clerical profession. AB - The concepts of work-to-family conflict and family-to-work conflict have been studied extensively in recent years. We propose a different means of understanding clergy work and family stressors because the boundaries between family and work are blurred within the clerical profession. We suggest, therefore, that the stressors associated with ordained ministry can be better analyzed if separated into two related, but distinct categories: (1) stressors stemming from the demands of the work (work-related stress) and (2) stressors stemming from the way the work impinges upon clerical family boundaries (boundary related stress). Utilizing the Pulpit and Pew Clergy Leadership Survey of 2001, we explore the association between these two forms of stress. Correlation analysis and simple and multiple regression models are used. A strong association between work-related stress and boundary-related stress is detected; length of time spent in the profession is related to reduced levels of boundary- and work related stress. PMID- 21647817 TI - Transaminitis: the lab test that has inflammation.... PMID- 21647821 TI - The adverse effects of flow-diverter stent-like devices on the flow pattern of saccular intracranial aneurysm models: computational fluid dynamics study. AB - BACKGROUND: Stent deployment across the aneurysmal neck has been established as one of the endovascular methods to treat intracranial aneurysms with or without coils. OBJECTIVE: The purpose is to study the possible adverse effects of deployment of the new flow-diverter stent-like devices (FD) on the flow characteristics of saccular aneurysm models. METHODS: Numerical simulations of the blood flow patterns in the artificial models of three aneurysms were studied. One model was designed without an FD stent, the second model with one FD stent, and the third model with two stents. Numerical simulation for incompressible laminar blood flow was conducted in the three artificial cerebral aneurysm models by means of computational fluid dynamics. RESULTS: There was a noticeable increase in the values of the circumferential pressure distributed on the walls of the aneurysm after stent deployment; this led to an increase the tension of the aneurysm surface and was considered to be an adverse effect. This pressure increase was further aggravated by the deployment of another stent. However, there is a beneficial effect of using FD stents, translating into the reduction of the flow velocity inside the aneurysm and wall shear stress at the inflow zone. This reduction decreases further with the deployment of another stent. CONCLUSION: Aneurysms become tenser after the deployment of one flow-diverter stent and (more tense still) after after the deployment of another stent. This principle should be kept in mind when choosing which group of aneurysms is the best candidate for such a treatment strategy. This study recommends deploying several FD stents during endovascular procedures until complete arrest of the blood flow occurs during the procedure; otherwise, the aneurysm may become tenser and dangerous if a slow blood flow jet still exists inside it at the end of the procedure. PMID- 21647818 TI - Sex differences in the recognition of emotional prosody in late childhood and adolescence. AB - We examined sex-related differences in the ability to recognize emotional prosody in late childhood (9-12 year olds) and adolescence (13-15 year olds) in relation to salivary testosterone levels. In order to examine both the accuracy and the sensitivity in labeling emotional prosody expressions, five intensities (20, 40, 60, 80, and 100%) for each of three emotion categories were used as stimuli. Totals of 25 male and 22 female children and 28 male and 28 female adolescents were tested on their recognition of happy, angry and sad prosody at the different intensities. The results showed that adolescent females were more sensitive to happy and sad prosody than males but not to angry prosody, whereas there were no sex-related differences in emotional prosody in late childhood for any of the emotional categories. Furthermore, salivary testosterone levels were higher in males than females in adolescence, but not in late childhood, suggesting that the sex differences for emotional prosody recognition emerges in adolescence during which testosterone levels become higher in males than females. PMID- 21647819 TI - Acute and population level toxicity of imidacloprid and fenpyroximate on an important egg parasitoid, Trichogramma cacoeciae (Hymenoptera: Trichogrammatidae). AB - One focus of integrated pest management (IPM) is the use of biological and chemical control in an optimal way. The availability of selective pesticides is important as is information about both lethal and sublethal effects of pesticides on biocontrol agents. Acute and sublethal effects of imidacloprid and fenpyroximate exposure were studied on adult stage of egg parasitoid Trichogramma cacoeciae Marchal and the emergence rate and life table parameters were determined. The adult wasps were exposed to field recommended concentration (FRC) of the pesticides on glass plates. Field rates of imidacloprid and fenpyroximate caused 100 and 32% adult mortality, respectively. Based on concentration-response experiments, the LC(50) values of imidacloprid and fenpyroximate were 6.25 and 1,949 ppm, respectively. The effect of imidacloprid and fenpyroximate on larvae, prepupae and pupae of the parasitoid was tested by exposing parasitized eggs of Sitotroga cerealella Olivier or Cydia pomonella L. to the FRC. Imidacloprid and fenpyroximate reduced adult emergence by 10.7 and 29%, respectively, when S. cerealella eggs were used as the host and 10.9 and 24.9%, respectively, when C. pomonella eggs were used as the host. Population parameters of emerged adults from treated pre-imaginal stages by FRC of the pesticides were also studied. The parameters were longevity and progeny production of emergent adults and also intrinsic rate of increase (r ( m )), generation time (T) and doubling time (DT). Longevity and progeny production of the emergent adults was not affected by pesticide exposure in comparison to the control. In addition, none of population parameters such as r ( m ), T and DT were affected by pesticide exposure. The intrinsic rate of increase for the control, fenpyroximate and imidacloprid exposed populations were 0.388, 0.374, and 0.372 female offspring per female per day, respectively. Overall, results of this study suggest a relative compatibility between fenpyroximate and T. cacoeciae, but imidacloprid showed deleterious effects on adults of the parasitoid. PMID- 21647820 TI - Single-item screens identified patients with elevated levels of depressive and somatization symptoms in outpatient physical therapy. AB - OBJECTIVE: Develop efficient and accurate screening tools to identify elevated levels of depressive or somatization symptoms, which can adversely affect functional status outcomes. METHODS: We conducted a secondary analysis of prospectively collected depressive and somatization symptoms (Symptom Checklist 90-Revised) data from 10,920 patients receiving outpatient physical therapy for a variety of neuromusculoskeletal diagnoses. Item response theory methods were used to analyze data, with particular emphasis on differential item functioning among groups of patients, and to identify potential screening items. Screening item accuracy for identifying patients with elevated symptoms was assessed with receiver-operating characteristic analyses. RESULTS: Seven items for depressive and 10 items for somatization symptoms represented unidimensional scales. Differential item functioning was negligible for demographic and clinical variables known to affect functional status outcomes. Items providing maximum information at the 88th percentile for depressive and 77th percentile for somatization scales accurately dichotomized patients into elevated versus not elevated symptom levels. CONCLUSIONS: Lack of differential item functioning suggested depressive and somatization screening could be useful in routine clinical practice and allowed the development of single-item screens that accurately identified patients with elevated depressive or somatization symptoms. Item response theory-based single-item screens may facilitate evaluation and management of heterogeneous populations receiving outpatient physical therapy. PMID- 21647822 TI - Development of basal endosperm transfer cells in Sorghum bicolor (L.) Moench and its relationship with caryopsis growth. AB - During sorghum caryopsis development, endosperm epidermal cells near the basal main vascular bundle are specialized by depositing wall ingrowths, differentiating into basal endosperm transfer cells (BETCs). All the BETCs together compose the basal endosperm transfer layer (BETL). BETCs are the first cell type to become histologically differentiated during endosperm development. The initiation and subsequent development of BETCs shows the pattern of temporal and spatial gradient. The developmental process of BETL can be divided into four stages: initiation, differentiation, functional, and apoptosis stage. A placental sac full of nutrient solutions would emerge, enlarge, and eventually disappear between the outmost layer of BETL and nucellar cells during caryopsis development. BETCs have dense cytoplasm rich in mitochondria, lamellar rough endoplasmic reticulum, Golgi bodies, and their secretory vesicles. They show a series of typical characteristics of senescence such as nuclei distortion and subcellular organelle deterioration during their specialization. BETCs probably play an active role in nutrient transfer into the starchy endosperm and embryo. The occurrence, development, and apoptosis of BETCs are in close relation to the caryopsis growth and maturation especially the enrichment of endosperm and the growth of embryo. The timing when BETL is fully developed, composed of three to four layers in radial direction and 70 to 80 rows in tangential direction, consists with the timing when average daily gain of caryopsis dry weight reaches its maximum. It is conceivable that measures that delay the senescence and death of BETCs would help to increase the crop yield. PMID- 21647823 TI - Development and evaluation of a small and mobile Magneto Alert Sensor (MALSE) to support safety requirements for magnetic resonance imaging. AB - OBJECTIVE: The purpose of this study is to (i) design a small and mobile Magnetic field ALert SEnsor (MALSE), (ii) to carefully evaluate its sensors to their consistency of activation/deactivation and sensitivity to magnetic fields, and (iii) to demonstrate the applicability of MALSE in 1.5 T, 3.0 T and 7.0 T MR fringe field environments. METHODS: MALSE comprises a set of reed sensors, which activate in response to their exposure to a magnetic field. The activation/deactivation of reed sensors was examined by moving them in/out of the fringe field generated by 7TMR. RESULTS: The consistency with which individual reed sensors would activate at the same field strength was found to be 100% for the setup used. All of the reed switches investigated required a substantial drop in ambient magnetic field strength before they deactivated. CONCLUSIONS: MALSE is a simple concept for alerting MRI staff to a ferromagnetic object being brought into fringe magnetic fields which exceeds MALSEs activation magnetic field. MALSE can easily be attached to ferromagnetic objects within the vicinity of a scanner, thus creating a barrier for hazardous situations induced by ferromagnetic parts which should not enter the vicinity of an MR-system to occur. PMID- 21647824 TI - Gender-specific association between ACE gene I/D polymorphism and blood pressure response to hydrochlorothiazide in Han Chinese hypertensive patients. AB - To evaluate the interaction between the angiotensin-converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism and gender with individual blood pressure response to hydrochlorothiazide (HCTZ) in hypertensives, we enrolled 829 mild moderate hypertensive patients. All subjects were given HCTZ (12.5 mg) orally each day for 6 weeks. A total of 776 patients completed the study. There was statistically significant interaction between the effects of genotype and gender on systolic (P = 0.002) and diastolic (P = 0.048) response after adjusting for covariables. Moreover, in each gender, the genotype that was associated with the greatest blood pressure response to HCTZ (DD homozygotes in men and II homozygotes in women) was also associated with the greatest increase in serum ACE activity in response to HCTZ. The results suggest that the I/D polymorphism of the ACE gene is associated with interindividual differences in the blood pressure response to a low dose of a diuretic in a gender-specific manner in the Han Chinese population with hypertension. PMID- 21647826 TI - [Skull base surgery in Germany. From microscope to endoscope and from radicality to functionality]. PMID- 21647825 TI - Hepatitis B virus X gene and hepatocarcinogenesis. AB - Chronic hepatitis B virus (HBV) infection has been identified as a major risk factor in hepatocellular carcinoma (HCC), which is one of the most common cancers worldwide. The pathogenesis of HBV-mediated hepatocarcinogenesis is, however, incompletely understood. Evidence suggests that the HBV X protein (HBx) plays a crucial role in HCC development. HBx is a multifunctional regulator that modulates transcription, signal transduction, cell cycle progression, apoptosis, protein degradation pathways, and genetic stability through interaction with host factors. This review describes the current state of knowledge of the molecular pathogenesis of HBV-induced HCC, with a focus on the role of HBx in hepatocarcinogenesis. PMID- 21647828 TI - [Interdisciplinary skull base surgery. From explorative crater cavity surgery to predictive tunnel surgery?]. AB - Total endoscopic or endoscopy-assisted surgical procedures are becoming more and more important for skull base surgery. In particular at the frontal skull base endoscopic procedures with planned opening of the subarachnoidal space have mostly replaced the microscope because of the good experiences in functional endoscopic sinus surgery (FESS). The advantages for the patients of endoscopic procedures with planned opening of the subarachnoidal space measured by criteria, such as mortality, morbidity or recurrence rate are unclear due to a lack of valid study data according to the criteria of evidence-based medicine. PMID- 21647827 TI - [Reconstruction of complex midfacial defects with individualized titanium implants]. AB - BACKGROUND: Loss of hard and soft tissue structures of the midface due to resection or trauma is associated with substancial functional and aesthetic deficits. Besides reconstruction of bony contours for preservation of orbit position and facial symmetry, reconstruction often requires simultaneous transplantation of soft tissue flaps for separation of nasal and oral cavities and refilling of soft tissue volume deficits. PATIENTS AND METHODS: A well established procedure of our institution will be demonstrated in 10 exemplary patients, in which titanium meshes are customized for individual defect situations using computer-assisted techniques in combination with soft tissue transfer if required. RESULTS: According to our experience, this procedure provides satisfactory results in functional as well as in aesthetic respects. Especially in patients with loss of bony structures of the orbit and preservation of orbital contents, this procedure forms optimal preconditions for prevention of enophthalmos and diplopia by preservation of the original orbital volume. CONCLUSION: Individualized titanium implants should be used more frequently in clinical routine for reconstruction of complex midfacial defects. PMID- 21647829 TI - [Modern skull base surgery from the perspective of neurosurgeons]. AB - At present, modern skull base surgery is a highly sophisticated interdisciplinary collaboration of various diagnostic and therapeutic disciplines. The overall goal is the treatment of complex tumorous, traumatic, vascular and inflammatory processes or developmental disorders of the skull base with preservation of function. The paper presents modern concepts, procedures and minimally invasive strategies in skull base surgery and also critically discusses the current trend to endoscopic and robot-assisted surgical techniques. PMID- 21647830 TI - [Neuroradiologic diagnostic and interventional procedures for diseases of the skull base]. AB - Besides image-guided biopsy techniques, the emphasis in the interdisciplinary cooperation between head and neck surgery and neuroradiology is on vessel occluding and preserving measures. Knowledge of dangerous anastomoses between extracranial and intracranial vessels is crucial. The principles of vessel occluding procedures including materials are presented and illustrated with case examples. Embolization of glomus tumors or epistaxis and preoperative permanent vessel occlusion techniques are demonstrated as well as vessel-preserving therapies, such as placement of covered stents for improving tumor resectability or after iatrogenic laceration of the internal carotid artery. PMID- 21647831 TI - [Surgical extirpation. The therapy of choice for chondrosarcoma of the larynx]. PMID- 21647832 TI - [Chondrosarcoma of the larynx. Experience gained in Freiburg since 1999]. AB - BACKGROUND: Chondrosarcomas are rare tumors of the head and neck. Nevertheless, they display the most common non-epithelial malignancy of the larynx. MATERIALS AND METHODS: Between 1999 and February 2010 we treated six patients with laryngeal chondrosarcoma. The group included two female and four male patients ranging in age from 54 to 82 years. RESULTS: An 82-year-old female patient died 3 months after diagnosis and tracheostomy due to other underlying diseases. An 82 year-old male patient underwent primary radiation therapy. In the other patients, we performed a modified hemilaryngektomy in three cases and a laryngectomy in one. In those four cases, there were no signs of recurrent disease 50, 85, 87 and 95 months after surgery, respectively. There were no local or distant metastases. CONCLUSIONS: Chondrosarcomas of the larynx are slow growing neoplasms. Metastases occur in less than 3% of cases. Complete resection is the therapy of choice. Function-preserving surgical approaches should be favoured. PMID- 21647833 TI - [Ultrasound microscopy in the upper aerodigestive tract. Initial clinical experiences]. AB - PURPOSE: Initial clinical experiences with ultrasound microscopy in the upper aerodigestive tract are described. MATERIALS AND METHODS: In the present pilot study, 20 healthy probands and 10 patients with lesions of the oral cavity, pharynx and larynx undergoing surgery were examined using a new prototype of an ultrasound microscope. RESULTS: A total of 24 normal and six pathological findings of the upper aerodigestive tract were assessed. These included normal mucosa of the floor of the mouth, the inner cheek, the palate and the vocal fold. In addition, invasive carcinoma of the floor of the mouth, the aryepiglottic fold and the vocal fold was found. Furthermore, a papilloma of the palatal arch and two epiglottic cysts were examined. CONCLUSION: Our results demonstrate that ultrasound microscopy is also viable in hollow organs. Pathological lesions differed clearly from normal mucosa. However, to recognize the regular pattern of different lesions of the upper aerodigestive tract, further investigations need to be carried out with a larger number of patients. PMID- 21647834 TI - [Auditory processing disorders--differential diagnosis. Guidelines of the German Society for Phoniatry and Pedaudiology]. AB - One prerequisite of diagnosing an auditory processing disorder (APD) is the differential diagnostic exclusion of language comprehension disorders, attention deficit hyperactivity disorders, cognitive impairment, as well as autistic-type diseases. This issue is discussed in detail in the updated guidelines for APD, as well as the logical consequences resulting thereof in terms of interpreting individual test results. This update is based in terms of content on the preceding guidelines of the German Society for Phoniatry and Pedaudiology (DGPP) and aligns itself closely with the Californian Speech-Language-Hearing Association, as well as guidelines of the American Academy of Audiology. PMID- 21647836 TI - Abstracts of the 10th World Congress on Inflammation. June 25-29, 2011. Paris, France. PMID- 21647835 TI - [Principles of cholesteatoma surgery]. AB - As a potentially life-threatening disease, cholesteatoma of the petrous bone generally requires a surgical approach. Surgery aims to produce an ear that is easy to care for and is free of recurrent or residual cholesteatoma. Hearing improvement is of secondary importance. Nevertheless the sound conduction system is generally reconstructed as a single-stage procedure. There are various possible methods of dealing with a cholesteatoma: (i) preserving or reconstructing the posterior meatal wall with an aerated mastoid (closed technique) or obliteration of the mastoid completely or partially after removal of the posterior wall (closed technique); and (ii) leaving the cavity open for inspection (open technique). In our opinion any technique where the mastoid is not open for inspection should be called a closed technique. Both techniques have their specific advantages and disadvantages. PMID- 21647837 TI - Combining NMR and X-ray crystallography in fragment-based drug discovery: discovery of highly potent and selective BACE-1 inhibitors. AB - Fragment-based drug discovery (FBDD) has become increasingly popular over the last decade. We review here how we have used highly structure-driven fragment based approaches to complement more traditional lead discovery to tackle high priority targets and those struggling for leads. Combining biomolecular nuclear magnetic resonance (NMR), X-ray crystallography, and molecular modeling with structure-assisted chemistry and innovative biology as an integrated approach for FBDD can solve very difficult problems, as illustrated in this chapter. Here, a successful FBDD campaign is described that has allowed the development of a clinical candidate for BACE-1, a challenging CNS drug target. Crucial to this achievement were the initial identification of a ligand-efficient isothiourea fragment through target-based NMR screening and the determination of its X-ray crystal structure in complex with BACE-1, which revealed an extensive H-bond network with the two active site aspartate residues. This detailed 3D structural information then enabled the design and validation of novel, chemically stable and accessible heterocyclic acylguanidines as aspartic acid protease inhibitor cores. Structure-assisted fragment hit-to-lead optimization yielded iminoheterocyclic BACE-1 inhibitors that possess desirable molecular properties as potential therapeutic agents to test the amyloid hypothesis of Alzheimer's disease in a clinical setting. PMID- 21647841 TI - Cationic bismuth-catalyzed hydroamination and direct substitution of the hydroxy group in alcohols with amides. AB - Bismuth-catalyzed hydroamination and direct substitution of the hydroxy group in alcohols are described in this chapter. Intermolecular 1:1 hydroamination of 1,3 dienes with carbamates, sulfonamides, and carboxamides was promoted by a combination of Bi(OTf)(3) and Cu(CH(3)CN)(4)PF(6). The mechanistic studies suggested that a cationic bismuth species would be an active species, which selectively promotes 1:1 hydroamination to give allylic amides in up to 96% yield. The cationic bismuth species was also applicable for hydroamination of vinyl arenes. The combination of Bi(OTf)(3) and KPF(6) was an excellent catalyst for direct substitution of the hydroxy group in allylic, propargylic, and benzylic alcohols with carbamates, sulfonamides, and carboxamides, giving allylic, propargylic, and benzylic amides, respectively, in up to 99% yield in one step. PMID- 21647840 TI - New applications for bismuth(III) salts in organic synthesis: from bulk chemicals to steroid and terpene chemistry. AB - Bismuth(III) salts are currently considered efficient and "ecofriendly" reagents and catalysts for the development of new applications in organic synthesis. The preparation of bismuth(III) triflate and its analogues is reviewed as well as some of their applications to the synthesis of bulk chemicals via electrophilic addition and cyclization reactions. The use of bismuth(III) salts in the development of new chemical processes involving steroids and terpenes as substrates is also discussed. PMID- 21647838 TI - Hsp90 inhibitors and drugs from fragment and virtual screening. AB - We have previously reported the structure-based optimisation of a number of series of potent compounds progressed as clinical candidates for oncology through inhibition of the ATPase activity of the molecular chaperone, Hsp90. The starting point for these candidates was compounds discovered using a combination of structure-based hit identification methods. This chapter summarises the overall story of how these methods were applied. Virtual screening of commercially available compounds identified a number of classes of compounds. At the same time, an initial fragment screen identified 17 fragments of various classes that bound to the N-terminal domain of Hsp90 with weak (0.5-10 mM) affinity. A subsequent screen identified a total of 60 compounds. This collection of fragments and virtual screening hits were progressed in a number of ways. Although two fragments could be observed binding together in the active site, the synthetic effort required to link these fragments was judged too high. For the resorcinol class of fragments, limited library synthesis generated compounds in the 1-10 MUM range. In addition, the resorcinol substructure was used to select commercially available compounds that were filtered using focussed docking in the Hsp90 active site to select further sets of compounds for assay. This identified structural motifs that were exploited during lead optimisation to generate AUY922, currently in Phase II clinical trials. In a separate campaign, features identified in the structures of fragments, evolved fragments and virtual screening hits bound to Hsp90 were combined to generate an oral series of compounds, progressed to preclinical candidates. The crystal structures were determined of many of the fragments bound to Hsp90 and provide examples of both maintenance and change of protein conformation on fragment binding. Finally, we analyse the extent to which our initial set of fragments recapitulates the key structural features of the Hsp90 inhibitors published to date. PMID- 21647839 TI - Synthesis of polypeptides by ring-opening polymerization of alpha-amino acid N carboxyanhydrides. AB - This chapter summarizes methods for the synthesis of polypeptides by ring-opening polymerization. Traditional and recently improved methods used to polymerize alpha-amino acid N-carboxyanhydrides (NCAs) for the synthesis of homopolypeptides are described. Use of these methods and strategies for the preparation of block copolypeptides and side-chain-functionalized polypeptides are also presented, as well as an analysis of the synthetic scope of different approaches. Finally, issues relating to obtaining highly functional polypeptides in pure form are detailed. PMID- 21647842 TI - Liver lead concentrations in raptors in New Jersey, USA, 2008-2010. AB - Lead exposure in New Jersey raptors was assessed by analyzing liver samples from carcasses obtained from wildlife rehabilitators. Samples were collected from 221 individuals representing 13 species. Concentrations were within the range of normal background exposure in 12 species. One red-tailed hawk had a liver lead concentration consistent with clinical poisoning (7.4 MUg/g wet weight), which represents an incidence of 1% (1/104) in that species and 0.5% (1/221) in the overall sample. A second red-tailed hawk had a liver lead concentration consistent with subclinical exposure (2.1 MUg/g wet weight). The combined incidence of elevated exposure (subclinical exposure + clinical poisoning) was 2% (2/104) in red-tailed hawks and 1% (2/221) in the overall sample. PMID- 21647843 TI - MicroRNAs in the human pituitary. AB - MicroRNAs (miRNAs) represent a novel class of small RNA molecules that play a crucial role as post-transcriptional regulators of gene expression. As evidence for the involvement of miRNAs in various cellular processes increases, it is important to examine how miRNAs regulate gene expression. In the pituitary, aberrant miRNA expression is strongly linked with neoplasia, thus suggesting they play a role in the control of cell proliferation in adenomas. Research has built fundamental connections between aberrant miRNA expression and clinicopathological features of pituitary adenomas. Moreover, deregulated expression of miRNA target genes is often implicated in important biological pathways and thus provides significant insight into the role of miRNAs in tumorigenesis. This review will assess the significance of miRNAs in pituitary pathology. PMID- 21647844 TI - Well-differentiated thyroid carcinoma with concomitant Hashimoto's thyroiditis present with less aggressive clinical stage and low recurrence. AB - Papillary thyroid carcinoma (PTC) and follicular thyroid carcinoma (FTC) are the most common differentiated thyroid cancers. Previous studies report that Hashimoto's thyroiditis (HT) concomitant with PTC is unusual and improves prognosis compared to classical PTC. Few previous studies address FTC concomitant with HT. In this study, we retrospectively analyzed data from one institution and compared clinical presentations and results of treatment of PTC and FTC with and without HT. In addition, studies comparing presentation and long term follow-up prognosis in classical PTC and FTC were conducted. A total of 1,788 PTC patients and 209 FTC patients underwent thyroidectomy with or without lymph node dissection and follow-up at Chang Gung Medical Center in Linkou, Taiwan. All thyroid carcinomas were pathologically classified according to World Health Organization criteria. Histological patterns of PTC were categorized as classical PTC, or PTC with HT. Follicular thyroid carcinoma patients were categorized as FTC or FTC with HT. The dataset contained a total of 1,703 PTC cases categorized as classical PTC, 85 cases of PTC with HT, 201 cases of FTC and eight cases of FTC with HT. Analysis of Classification of Malignant Tumors (TNM) stage revealed a higher percentage of classical PTC in stage IV than HT group (12.03% vs. 4.70%). Mean tumor size of classical PTC was larger than HT group. Although 42.3% of FTC cases presented with distant metastases, no cases of FTC with HT presented with distant metastasis. Cancer-specific mortality was higher in classical PTC group than in PTC with HT. There was 53.2% of FTC without HT assigned recurrent status, and six of them died of thyroid cancer. No cancer mortality or recurrence in HT with FTC. PTC and FTC with HT presented with better clinical stage and better prognosis after same therapeutic modality. In conclusions, both PTC and FTC with HT have less aggressive clinical presentation and better prognosis. PMID- 21647846 TI - Topical anesthetic EMLA for postoperative wound pain in stereotactic gamma knife radiosurgery: a perspective, randomized, placebo-controlled study. AB - BACKGROUND: Patients who undergo stereotactic gamma knife radiosurgery (GKRS) need a rigid frame fixation for the stereotactic procedures. Many patients suffered from postoperative wound pain after frame removal. The present study investigated whether an additional application of a topical anesthetic prior to frame removal could reduce this discomfort. PATIENTS AND METHODS: 60 patients who underwent GKRS were enrolled in this study. Of these 60 patients, 30 were treated with a topical application of EMLA, a eutectic mixture of 2.5% lidocaine and 2.5% prilocaine; the remaining 30 were treated with a placebo. The nurses explained the definition of the visual analogue scale (VAS, scored from 0 to 10), and the patients evaluated their pain at 7 time points during the GKRS procedure by using the VAS. After each of these evaluations, the patients' vital signs (blood pressure, heart rate, and respiratory rate) were measured. RESULTS: There was no significant difference in the patients' age, gender, duration of frame fixation, and types of the lesions between the EMLA and placebo groups. The EMLA group reported significantly lower pain scores 20 and 60 min after frame removal than the placebo group (p=0.001 and p<0.001, respectively). Additionally, patients in the placebo group had significantly higher blood pressure readings compared with baseline data, during and after frame removal, thus indicating that postoperative wound pain caused them more discomfort after frame removal. CONCLUSION: EMLA when applied 60 min before frame removal has an anesthetic effect of reducing the postoperative wound pain in patients who undergo GKRS. PMID- 21647847 TI - Acute encephalopathy in a patient with Dravet syndrome. AB - Dravet syndrome (severe myoclonic epilepsy in infancy) is an epileptic syndrome with various types of seizures that begin in the first year of life and may result in intellectual impairment. Mutations of the SCN1A gene are the most prevalent genetic cause of Dravet syndrome. In this study, we report a 12-year old girl with Dravet syndrome carrying an SCN1A mutation, c.2785Cdel (L929del fsX934). She had an episode of status epilepticus and persistent lethargy after 48 h of acute febrile illness that was preceded by an annual flu vaccination. Low voltage activities detected by electroencephalogram and elevated neuron-specific enolase/interleukin-6 concentrations in the cerebrospinal fluid suggested acute encephalopathy. MRI showed abnormalities in the bilateral thalami, cerebellum and brainstem. These abnormalities were protracted over a month. The biochemical and MRI characteristics of this case are different from any known type of encephalopathy, and may suggest a vulnerability of neurons expressing mutant SCN1A in the brain. PMID- 21647845 TI - Effects of high-frequency oscillatory ventilation on systemic and cerebral hemodynamics and tissue oxygenation: an experimental study in pigs. AB - BACKGROUND: In this study, we compare the effects of high frequency oscillatory ventilation (HFOV) with those of lung-protective volume-controlled ventilation (VCV) on cerebral perfusion, tissue oxygenation, and cardiac function with and without acute intracranial hypertension (AICH). METHODS: Eight pigs with healthy lungs were studied during VCV with low tidal volume (V(T): 6 ml kg(-1)) at four PEEP levels (5, 10, 15, 20 cm H(2)O) followed by HFOV at corresponding transpulmonary pressures, first with normal ICP and then with AICH. Systemic and pulmonary hemodynamics, cardiac function, cerebral perfusion pressure (CPP), cerebral blood flow (CBF), cerebral tissue oxygenation, and blood gases were measured after 10 min at each level. Transpulmonary pressures (TPP) were calculated at each PEEP level. The measurements were repeated with HFOV using continuous distending pressures (CDP) set at TPP plus 5 cm H(2)O for the corresponding PEEP level. Both measurement series were repeated after intracranial pressure (ICP) had been raised to 30-40 cm H(2)O with an intracranial balloon catheter. RESULTS: Cardiac output, stroke volume, MAP, CPP, and CBF were significantly higher during HFOV at normal ICP. Systemic and cerebral hemodynamics was significantly altered by AICH, but there were no differences attributable to the ventilatory mode. CONCLUSION: HFOV is associated with less hemodynamic compromise than VCV, even when using small tidal volumes and low mean airway pressures. It does not impair cerebral perfusion or tissue oxygenation in animals with AICH, and could, therefore, be a useful ventilatory strategy to prevent lung failure in patients with traumatic brain injury. PMID- 21647850 TI - [Predictors of participation in medical rehabilitation follow-up in working patients with chronic back pain]. AB - BACKGROUND AND AIM: Outpatient follow-up programmes aim to increase the sustainability of rehabilitation. However, the factors influencing participation in follow-up programmes are still unclear. The aim of this study was to examine participation in the MERENA follow-up programme of the Rhineland-Palatinate German Pension Fund. METHOD: The study was conducted in 12 outpatient rehabilitation centres with 192 working patients who were being treated for chronic back pain at the time the survey was conducted. Both patients and physicians completed a written survey at the outset of rehabilitation, on completion, and again (if applicable) at the end of the follow-up programme. The data collected mainly concerned the patients' health and factors related to their occupational situation (e. g. functional capacity, working capacity) and reasons for refusing to participate in the MERENA follow-up programme. Predictors for participation in the follow-up programme were determined using binary logistic regression analysis. RESULTS: On completion of rehabilitation, nearly all patients were given the recommendation to participate in the follow-up programme. Half of these patients took advantage of the programme. The most frequently given reason for refusal to participate was that participation in the programme was not compatible with their duties at work. Low functional capacity and continued work disability increased the probability that a patient would take part in a follow up programme after rehabilitation. In contrast, a longer commute to the centre was an obstacle to participation. Women were more likely to participate in the programme than men. CONCLUSIONS: The results indicate that participation in a follow-up programme is often not compatible with employment. We could not satisfactorily explain why women were more likely to participate in the programme. This result could have been related to women's more flexible time schedules. An improvement of the current situation could be achieved by having follow-up programmes closer to the home, by flexible follow-up offerings, alternative follow-up services (e. g. in certified physiotherapy centres and sport clubs) as well as by integrating companies in follow-up planning and implementation. PMID- 21647849 TI - [What are the causes of death in patients with spinal cord injury today?--a descriptive analysis of 102 cases]. AB - As there are only few reliable data concerning mortality of SCI patients, this retrospective monocentric cohort study was carried out. Despite essential improvements in intensive medical care from the accident scene to clinic life, comprehensive rehabilitation, and implementation of a lifelong aftercare system, the life expectancy of SCI patients is still reduced. Especially patients with high tetraplegia die significantly earlier from pulmonary complications. The longer the onset of SCI is survived, the more patients die from age-related diseases. In old paraplegic patients, pressure sores are the only major SCI related complication. Successful social reintegration and professional care are the most important factors for an expanded lifespan after occurrence of a SCI. Hence, the special impact of lifelong treatment of SCI patients ("comprehensive care") is confirmed. PMID- 21647848 TI - The plasminogen activator inhibitor-1 gene polymorphism in determining the risk of pediatric ischemic stroke--case control and family-based study. AB - Pediatric ischemic stroke, though relatively rare, remains an important medical problem since 20-40% of patients have recurrent strokes and 50-85% of them suffer from long-term neurological deficits. Approximately 20-50% of the affected children have prothrombotic disorders, therefore upon looking for possible genetic causes of the disease we focused on the plasminogen activator inhibitor (PAI-1)--the major inhibitor of fibrinolysis. The aim of the present study was to investigate a possible association between the -675_-674insG PAI-1 gene polymorphism and pediatric ischemic stroke. The study population consisted of 343 individuals: 70 children with ischemic stroke, 140 their biological parents and 133 control children. The PAI-1 gene polymorphism was genotyped using the restriction fragment length polymorphism and was visualized by AgNO3 staining. The transmission/disequilibrium test showed exactly the same transmission of alleles from parents to the affected children (37:37). The case-control model also did not reveal any statistical significance in alleles and genotypes distribution between patients and control children. The obtained results suggest that the 4 G/5 G polymorphism of the PAI-I gene is not a risk factor of ischemic stroke in Polish children. PMID- 21647851 TI - [The development of an instrument assessing the relevance of rehabilitation outcomes for patient participation: retest reliability and descriptive results]. AB - OBJECTIVE: In the context of an increasing patient orientation in health care, surveying patients on the success of treatment has become highly relevant. The objective of the study was to develop and test an instrument for surveying the patients' perceptions of the participation relevance of changes in health status after inpatient medical rehabilitation with respect to various areas of life for 3 diagnoses: breast cancer, chronic ischemic heart disease, and chronic back pain. METHODS: The instrument refers to certain disease-specific areas of health (e. g., mobility) with regard to which the patients are asked whether they perceived an improvement and if so, what improvement in participation had ensued as a result. A questionnaire study (measuring time points: end of rehab and 6 months after the end of rehab) was conducted in 3 samples (breast cancer: n=297, chronic ischemic heart disease: n=295, chronic back pain: n=186) to test the methodology. The share of missing values and undesirable ceiling/floor effects were assessed to analyze the distribution characteristics. In a partial sub sample, a third measuring time point was introduced approx. 10 days after the second measuring time point, and the retest reliability of the assessment of participation relevance is determined using the ICC. We also present descriptive results relative to the participation relevance from the patient perspective. RESULTS: The instrument shows good distribution characteristics for the items. Regarding retest reliability, there were satisfactory values for all diagnoses in all areas of health. The descriptive analyses showed that patients who perceived a noticeable improvement in their health status at the end of rehabilitation or 6 months after inpatient rehab generally assessed the participation relevance of this improvement in health to be "moderate" to "strong". DISCUSSION: The concept of assessing the participation relevance from the patient perspective is a satisfactorily reliable construct. By including the subjective significance of treatment success it takes into account the concept of clinical significance from the patient's perspective, a concept which has thus far been considered only insufficiently. Results show that patients ascribe the health improvements achieved by rehabilitation a noticeable relevance for their participation. PMID- 21647852 TI - [The factor structure of direct and indirect methods for measuring change in medical rehabilitation--analyses on item level]. AB - AIM OF THE STUDY: The aim of the study was to examine the direct and the indirect change measurement; previous studies comparing the 2 methods demonstrated that they agree only to a limited extent. Analyses of the factorial structure have shown that indicators of direct change measurement were assigned regardless of the content of a dimension common to them. This paper examined whether previous results obtained on scale level were also valid on item level. METHODS: Empirical data were obtained from 3 rehabilitation patient samples (n=466, n=194, n=610). The factorial structures of the direct and indirect measurements of change were assessed and compared using principal components analysis with oblique rotation. Additionally, analyses were performed stratified across clinical indications and only for items of the direct change measurement. RESULTS: The factorial structure of 3 data sets showed that the previously published scale-level results were also valid on item level. Analyses demonstrated only low to medium agreement between the 2 methods of change measurement. The indicators of direct change measurement loaded regardless of their content on one or a few common dimensions, which did not agree with those of the indirect change measurement. CONCLUSION: In considering the 2 methods of measuring change, it is apparent that the standard method of change measurement (indirect change measurement) is suitable to reflect a comprehensive and multi-dimensional representation of changes within the field of medical rehabilitation evaluation. PMID- 21647853 TI - [Rehabilitation from the patient perspective--a qualitative analysis of the treatment beliefs of patients in inpatient orthopedic and psychosomatic rehabilitation]. AB - PURPOSE: As the common sense self-regulation model (CSM) was further developed in the late 1990s, its focus was, along with subjective illness perceptions, the treatment representations of patients, in particular medication-related perceptions. However until now, only few studies have dealt with subjective concepts regarding non-medication treatment. The objective of this study was to explore the core content areas of the treatment concept of rehabilitation patients as a basis for developing a questionnaire to survey rehabilitation related treatment beliefs. METHODS: In 5 inpatient rehabilitation centres for psychosomatic and musculoskeletal diseases, guided focus groups were conducted with a total of n=25 rehabilitation patients. Some 56% of the participants were female; the average age was 52.8 years (SD=10.8). The interviews were transcribed and analyzed using the computer programme Atlas.ti based on the method recommended by Mayring in a multi-stage qualitative content analysis procedure. RESULTS: In the analysis of the discussion transcriptions, a total of 579 patient statements were categorized with 49 codes that were assigned to the areas (1) expectations and beliefs with respect to the rehabilitation process, (2) expectations of results, and (3) fears associated with rehabilitation. The process expectations were the most frequently discussed topic, constituting 58.2% of all patient statements. It was found that the patients associate not only numerous individual treatments with rehabilitation but also have concrete perceptions about the specific form rehabilitation should have, their own role in the rehabilitation process, and features of the overall rehabilitation concept. The outcome expectations are related to the areas activities of daily life and job, psyche, soma, and effectiveness of rehabilitation. Fears with respect to rehabilitation generally played only a subordinate role for those surveyed. The indication-specific analyses showed that the perceptions regarding the realization and form of rehabilitation differed among the various diagnosis groups, but the patients named similar categories for expectations of results and fears, with only some variation in importance. CONCLUSION: The focus groups allowed good insights into the patient perspectives of rehabilitation. Simultaneously, a good basis was created for generating contents of items for a questionnaire on the rehabilitative treatment concept, so that along with literature analyses, the qualitative method proved to be a suitable approach and good source for developing a questionnaire. PMID- 21647854 TI - [Ambulant vocational rehabilitation of persons with mental illness]. AB - The composition of participants and the success of rehabilitation of an ambulant vocational rehabilitation programme for persons with mental illness was examined (Integration Seminar; 1994-2009; n=703). The programme applies an individualized concept of rehabilitation. The findings show that average age and share of participants with psychotic illness had increased over the years. The share of long-term unemployed persons in all was very high and had increased strongly in the 2002-2005 period. Depending on the year of programme completion, the share of successful participants ranged between 50% and 60% a year. The success of rehabilitation turned out relatively stable over all measurement times (r (s)=0.52-0.83) and correlates with resilience, age and duration of unemployment. This implies the necessity of an early beginning of vocational rehabilitation, as the probability of success decreases with increasing duration of prior unemployment. Overall, the concept of individualized ambulant vocational rehabilitation turns out very successful. PMID- 21647855 TI - [Goal setting in inpatient medical rehabilitation: exploring the current practice]. AB - OBJECTIVE: Goal setting is regarded as a key element in the rehabilitation process. Information about current goal setting practice is limited. However, there is potential for further improvement, e. g. as far as patient participation in the goal setting process is concerned. The study reports results of a survey on the current practice of goal setting in medical rehabilitation, focusing on development potential. The study examines acceptance and requirements for goal setting as well as patients' and professionals' experiences regarding goal setting during the rehabilitation process, with an emphasis on patient participation. METHODS: A total of n=40 rehabilitation professionals and n=210 inpatients with a diagnosis of chronic back pain, diabetes mellitus type 2 or coronary heart disease were surveyed at six rehabilitation centres using a questionnaire. Questionnaires contained predominantly closed-ended items regarding the current practice of goal setting. Several items were identical for both groups. In addition to the quantitative evaluation by means of frequency analyses free text data were evaluated. RESULTS: Rehabilitation professionals saw various benefits in goal setting with patients. Many professionals experienced patients to have personal goals for their rehabilitation, and most of them believed that patient participation in goal setting is possible. Also, barriers and requirements for goal setting were identified. A consistent concept for goal setting, involvement of and exchange between the different occupational groups and an elaborate handling of information in the context of goal setting seem to be realised at least partially. Professionals and patients reported similar frequencies of talks on goal setting at different times during the rehabilitation course. Patient participation seems to be realised to some extent but not comprehensively. The results show slight discrepancies between patients' and professionals' statements. DISCUSSION: Current practice of goal setting in inpatient rehabilitation of patients with the diagnoses named above is altogether on a relatively advanced level in the rehabilitation centres taking part in the study. The study identifies potential for development concerning a comprehensive implementation of goal setting, the realisation of patient participation, goal documentation and their integration into rehabilitation. Moreover, deeper involvement of the rehabilitation team and a stronger conceptual integration of the subject within the centres seem desirable. This can be taken up within the scope of interventions. PMID- 21647856 TI - Expanding refractory rectus sheath hematoma: a therapeutic dilemma. AB - Rectus sheath hematoma is an uncommon but well-described complication of a tussive paroxysm. It is an accumulation of blood within the sheath of the rectus abdominis secondary to disruption of the epigastric vessels or the rectus muscle and is often misdiagnosed as acute abdomen. Increases in the number of elderly patients and the use of therapeutic anticoagulation may increase the prevalence and severity of rectus sheath hematomas encountered in clinical practice. Expanding rectus sheath hematomas are occasionally refractory to conservative treatment and may require hemostatic intervention. Here, we describe the case of an 87-year-old woman who presented with two separate rectus sheath hematomas that were precipitated by a paroxysm of coughing. Repeated computed tomography showed two separate expanding rectus sheath hematomas, which were not accompanied by obvious contrast extravasation on angiography. Empiric left inferior epigastric artery embolization resulted in rapid hemodynamic stabilization, and the hematomas shrank gradually. Early empiric transcatheter arterial embolization may be appropriate for patients who are poor surgical candidates and have enlarging hematomas that are refractory to conservative treatment. PMID- 21647857 TI - The relationship between the myocardial T2* value and left ventricular volumetric and functional parameters in thalassemia major patients. AB - PURPOSE: Cardiac involvement in thalassemia major (TM) is mainly characterized by left ventricular dysfunction caused by iron overload. Cardiovascular magnetic resonance imaging (MRI) including myocardial T2* measurement is becoming increasingly popular for quantitatively evaluating myocardial iron overload. The aim of this study was to evaluate the relationship between the myocardial T2* value and left ventricular functional parameters and to examine the associations between the degree of cardiac iron load and various clinical parameters. MATERIALS AND METHODS: A retrospective analysis of 47 patients (25 males and 22 females; mean age, 23.0+/-5.4 years) with TM was performed. Myocardial iron load was assessed by T2* measurements, and volumetric functions were analyzed using the steady state free precession sequence. RESULTS In patients with myocardial iron deposition (T2* < 20 ms), the mean left ventricular ejection fraction (LVEF) was 64.73+/-4.94%. The LVEF of patients with myocardial siderosis was significantly lower than that of patients without myocardial siderosis (r=0.35, P = 0.014). Inverse and significant correlations between both the left ventricular (LV) end-systolic volume index and the LV end-diastolic volume index and the myocardial T2* value (r=-0.32, P = 0.027 and r=-0.29, P = 0.046, respectively) were observed. There was an inverse correlation between the myocardial T2* value and the liver iron concentration (r=-0.31, P = 0.037). Cardiac T2* was not associated with serum ferritin levels, pre-transfusion hemoglobin levels or the annual red cell consumption rate. CONCLUSION: Myocardial iron load assessed by cardiac MRI (T2*) is associated with deterioration in left ventricular function. Thalassemia major patients with myocardial siderosis may have LVEF values within normal limits, but this result must be interpreted cautiously. PMID- 21647858 TI - Bias associated with mining electronic health records. AB - Large-scale electronic health record research introduces biases compared to traditional manually curated retrospective research. We used data from a community-acquired pneumonia study for which we had a gold standard to illustrate such biases. The challenges include data inaccuracy, incompleteness, and complexity, and they can produce in distorted results. We found that a naive approach approximated the gold standard, but errors on a minority of cases shifted mortality substantially. Manual review revealed errors in both selecting and characterizing the cohort, and narrowing the cohort improved the result. Nevertheless, a significantly narrowed cohort might contain its own biases that would be difficult to estimate. PMID- 21647860 TI - Quantitative magnetic resonance imaging evidence of synovial proliferation is associated with radiographic severity of knee osteoarthritis. AB - OBJECTIVE: To evaluate the relationships between both quantitative and semiquantitative assessments of the degree of knee synovitis on 3T magnetic resonance imaging (MRI) and the severity of knee osteoarthritis (OA) on radiography. METHODS: Fifty-eight patients with knee OA underwent nonfluoroscopic fixed-flexion knee radiography. In addition, dynamic contrast-enhanced 3T MRI of the knees was performed, before and after gadolinium administration, to quantify synovial membrane volume (SV) as a measure of synovial proliferation (expressed as the quantitative SV), and semiquantitative measures of synovitis were also applied using both contrast-enhanced and unenhanced images. Two radiologists scored the knee radiographs using the Osteoarthritis Research Society International atlas; interreader agreement was assessed using kappa statistics and concordance correlation coefficients. Multiple linear and logistic regression analyses were used to assess associations among variables, while controlling for the effects of age, body mass index, sex, and meniscal extrusion. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated for measures of disease activity. RESULTS: The Kellgren/Lawrence (K/L) grade of radiographic knee OA severity (beta=0.78), the diseased compartment joint space width (dcJSW) (beta=-0.22), and the diseased compartment joint space narrowing (dcJSN) score (beta=0.53) were each significantly associated with the quantitative SV (P=0.0001, P=0.0003, and P=0.0001, respectively). Furthermore, the quantitative SV strongly correlated with the total volume of subchondral bone marrow lesions (BMLs) (beta=0.22, P=0.0003). The K/L grade, dcJSW, and dcJSN score were each significantly associated with the semiquantitative Boston Leeds Osteoarthritis Knee Score (BLOKS) for the extent of infrapatellar synovitis (OR 9.05 [95% CI 1.94, 42.3] for K/L grade; OR 0.75 [95% CI 0.54, 1.03] for dcJSW; and OR 2.22 [95% CI 1.15, 4.31] for dcJSN score) and extent of joint effusion (OR 5.75 [95% CI 1.23, 26.8] for K/L grade; OR 0.70 [95% CI 0.50, 0.98] for dcJSW; and OR 1.96 [95% CI 1.02, 3.74] for dcJSN score). In addition, the semiquantitative synovitis grade on contrast-enhanced MRI was significantly associated with the K/L grade (beta=0.036, P=0.0040) and dcJSN score (beta=0.015, P=0.0266), and also significantly associated with the BLOKS synovitis score. CONCLUSION: Synovitis is a characteristic feature of advancing knee OA and is significantly associated with the K/L grade, JSW, JSN score, and total volume of BMLs on radiographs. Furthermore, BLOKS scoring of synovitis on unenhanced MRI is associated with measurements of synovitis on contrast-enhanced MRI. PMID- 21647862 TI - Clinical images: Extensive calcinosis in lupus panniculitis. PMID- 21647861 TI - Morphologic differences between the hips of Chinese women and white women: could they account for the ethnic difference in the prevalence of hip osteoarthritis? AB - OBJECTIVE: Hip osteoarthritis (OA) is a common disabling disease, which has a much higher prevalence in whites than in Asians. The reasons for this ethnic difference in prevalence are unknown. Hip OA is often thought to be secondary to morphologic abnormalities. This study was undertaken to examine whether particular abnormalities predisposing to hip OA occur more frequently in whites and whether these differences in hip shape account for differences in the prevalence of OA. METHODS: A morphometric study was performed on 400 hips of 200 female participants without OA from 2 studies, the Beijing OA Study and the Study of Osteoporotic Fractures from the US. We focused on measures of hip dysplasia and impingement (lateral center-edge angle, impingement angle, acetabular slope, femoral head-to-femoral neck ratio, and the crossover sign) and compared data from the hips of Chinese and white women. RESULTS: Compared with their Chinese counterparts, white women had a lower mean impingement angle (83.6 degrees versus 87.0 degrees ; P=0.03) and were more likely to have center-edge angles suggestive of impingement (>35 degrees ; 11% of hips in Chinese versus 23% of hips in whites, P=0.008). In contrast, low center-edge angles suggesting dysplasia (<20 degrees ) were found more often in Chinese women (22% of hips in Chinese versus 7% of hips in whites, P=0.005). CONCLUSION: In a study of elderly women without signs of OA, the morphometry of impingement and asphericity was more common in the hips of white women compared with Chinese women. Our findings suggest that whites may be at higher risk of hip OA than Chinese because of morphologic findings that predispose whites to femoroacetabular impingement. PMID- 21647863 TI - Role of mesenchymal stem cells in reestablishing immunologic tolerance in autoimmune rheumatic diseases. PMID- 21647864 TI - Therapeutic approaches in the treatment of juvenile dermatomyositis in patients with recent-onset disease and in those experiencing disease flare: an international multicenter PRINTO study. AB - OBJECTIVE: To evaluate response to therapy over a 24-month period in a large prospective international cohort of patients with juvenile dermatomyositis (DM). METHODS: The study included 145 patients with recent-onset juvenile DM and 130 juvenile DM patients experiencing disease flare, all of whom were <18 years old. Disease activity parameters and therapeutic approaches in 4 geographic areas were analyzed at baseline and at 6, 12, and 24 months. Response was assessed according to the Pediatric Rheumatology International Trials Organization (PRINTO) juvenile DM response criteria, and data were reported "as observed" and in the intent-to treat (ITT) population. RESULTS: Patients with recent-onset juvenile DM at baseline had higher baseline disease activity and greater improvement over 24 months when compared to juvenile DM patients experiencing disease flare at baseline. Methotrexate (MTX) or high-dose corticosteroids were administered more frequently to patients with recent-onset juvenile DM, compared to juvenile DM patients experiencing disease flare, who were more likely to receive cyclosporine. Compared to patients from Western and Eastern Europe, a higher proportion of patients from South and Central America and North America received pulse steroids, and the average steroid dosage was higher in the North American and South and Central American patients. The use of MTX was similar in all 4 regions, while cyclosporin A was more frequently used in Western Europe. In the "as observed" analysis, 57.9% of the patients with recent-onset juvenile DM and 36.4% of the patients experiencing disease flare (P<0.001) reached at least a 70% response by PRINTO criteria at 6 months; these proportions had increased at month 24 to 78.4% and 51.2%, respectively (P<0.001). Corresponding results of the ITT analysis were much lower, with only one-third of the patients able to maintain the initial assigned therapy over 24 months. CONCLUSION: Patients with recent onset juvenile DM are more likely to achieve significant clinical improvement over 24 months, when compared to patients experiencing flares of juvenile DM. Internationally, various therapeutic approaches are used to treat this disease. PMID- 21647865 TI - Cigarette smoking is not a risk factor for systemic sclerosis. AB - OBJECTIVE: To investigate the association of cigarette smoking with susceptibility to systemic sclerosis (SSc) in a large, well-defined patient population. METHODS: We conducted a review of 1,379 patients with SSc enrolled in the Scleroderma Family Registry and DNA Repository and/or the Genetics versus Environment in Scleroderma Outcome Study cohort. Smoking history was obtained from chart review or via telephone interview. Patients with SSc were subsequently categorized as never smokers or ever smokers. Patients with SSc for whom smoking data were available were matched 2:1 by age, sex, ethnicity, and state of residence to control subjects, using the Behavioral Risk Factor Surveillance System. RESULTS: The majority of patients were white (74.2%), with Hispanics and blacks representing 11.3% and 9.7%, respectively. Most patients had limited cutaneous involvement (54%). For our comparative analyses, 621 patients were matched with control subjects. There was no significant difference in age, sex, ethnicity, and SSc subtype between matched versus unmatched patients. The majority of patients had never smoked (57%), while 43% of patients were classified as ever smokers. The patients with SSc did not differ from control subjects in terms of their smoking behavior (odds ratio [OR] 1.020, 95% confidence interval [95% CI] 0.839-1.240, P=0.842). Anti-topoisomerase I antibody positive patients were more likely to be never smokers (OR 0.648, 95% CI 0.421 0.998, P=0.049), whereas no such association was observed with anticentromere and anti-RNA polymerase III antibodies. CONCLUSION: Unlike its role in rheumatoid arthritis, smoking does not confer a risk for development of SSc, although it may impact disease severity. PMID- 21647866 TI - Characterization of interleukin-7 and interleukin-7 receptor in the pathogenesis of rheumatoid arthritis. AB - OBJECTIVE: To characterize the expression of interleukin-7 (IL-7) and IL-7 receptor (IL-7R) in rheumatoid arthritis (RA) synovial tissue and to examine their regulation and pathogenic role in macrophages, endothelial cells, and synovial tissue fibroblasts in RA. METHODS: Expression of IL-7 and IL-7R in RA and normal synovial tissue was demonstrated by immunohistochemistry. Expression and regulation of IL-7 and IL-7R in RA peripheral blood in vitro-differentiated macrophages, RA synovial tissue fibroblasts, and human microvascular endothelial cells (HMVECs) were determined by real-time reverse transcription-polymerase chain reaction and/or flow cytometry. Enzyme-linked immunosorbent assay was used to examine production of proangiogenic factors by IL-7-activated macrophages, RA fibroblasts, and endothelial cells. RESULTS: IL-7 and IL-7R were coexpressed on RA synovial tissue lining and sublining macrophages and endothelial cells. Expression of IL-7 and its receptor was significantly elevated in RA synovial fluid and peripheral blood macrophages as well as RA fibroblasts, compared to normal cells. Toll-like receptor 4 ligation (with lipopolysaccharide) and tumor necrosis factor alpha (TNFalpha) stimulation modulated expression of IL-7 and IL 7R on RA macrophages and HMVECs. However, in RA fibroblasts, lipopolysaccharide and TNFalpha activation increased expression of IL-7R only. IL-7 also mediated RA pathogenesis by inducing production of potent proangiogenic factors from macrophages and endothelial cells. CONCLUSION: We have identified, for the first time, regulators of IL-7 and IL-7R expression in RA fibroblasts, RA peripheral blood in vitro-differentiated macrophages, and endothelial cells. Our results document a novel role of IL-7 in RA angiogenesis. PMID- 21647867 TI - Implementation of a treat-to-target strategy in very early rheumatoid arthritis: results of the Dutch Rheumatoid Arthritis Monitoring remission induction cohort study. AB - OBJECTIVE: Clinical remission is the ultimate therapeutic goal in rheumatoid arthritis (RA). Although clinical trials have proven this to be a realistic goal, the concept of targeting at remission has not yet been implemented. The objective of this study was to develop, implement, and evaluate a treat-to-target strategy aimed at achieving remission in very early RA in daily clinical practice. METHODS: Five hundred thirty-four patients with a clinical diagnosis of very early RA were included in the Dutch Rheumatoid Arthritis Monitoring remission induction cohort study. Treatment adjustments were based on the Disease Activity Score in 28 joints (DAS28), aiming at a DAS28 of <2.6 (methotrexate, followed by the addition of sulfasalazine, and exchange of sulfasalazine with biologic agents in case of persistent disease activity). The primary outcome was disease activity after 6 months and 12 months of followup, according to the DAS28, the European League Against Rheumatism (EULAR) response criteria, and the modified American College of Rheumatology (ACR) remission criteria. Secondary outcomes were time to first DAS28 remission and outcome of radiography. RESULTS: Six-month and 12-month followup data were available for 491 and 389 patients, respectively. At 6 months, 47.0% of patients achieved DAS28 remission, 57.6% had a good EULAR response, and 32.0% satisfied the ACR remission criteria. At 12 months, 58.1% of patients achieved DAS28 remission, 67.9% had a good EULAR response, and 46.4% achieved ACR remission. The median time to first remission was 25.3 weeks (interquartile range 13.0-52.0). The majority of patients did not have clinically relevant radiographic progression after 1 year. CONCLUSION: The successful implementation of this treat-to-target strategy aiming at remission demonstrated that achieving remission in daily clinical practice is a realistic goal. PMID- 21647868 TI - Impact of ineffective oesophageal motility and wrap type on dysphagia after laparoscopic fundoplication. AB - BACKGROUND: Laparoscopic 360 degrees fundoplication is the most common operation for gastro-oesophageal reflux disease, but is associated with postoperative dysphagia in some patients. Patients with ineffective oesophageal motility may have a higher risk of developing postoperative dysphagia, but this remains unclear. METHODS: From 1991 to 2010, 2040 patients underwent primary laparoscopic fundoplication for gastro-oesophageal reflux disease and met the study inclusion criteria; 343 had a 90 degrees , 498 a 180 degrees and 1199 a 360 degrees fundoplication. Primary peristalsis and distal contraction amplitude during oesophageal manometry were determined for 1354 patients. Postoperative dysphagia scores (range 0-45) were recorded at 3 and 12 months, then annually. Oesophageal dilatations and/or reoperations for dysphagia were recorded. RESULTS: Preoperative oesophageal motility did not influence postoperative dysphagia scores, the need for dilatation and/or reoperation up to 6 years. Three-month dysphagia scores were lower after 90 degrees and 180 degrees compared with 360 degrees fundoplication (mean(s.e.m.) 8.0(0.6) and 9.8(0.5) respectively versus 11.9(0.4); P < 0.001 and P = 0.003), but these differences diminished after 6 years of follow-up. The incidence of dilatation and reoperation for dysphagia was lower after 90 degrees (2.6 and 0.6 per cent respectively) and 180 degrees (4.4 and 1.0 per cent) fundoplications than with a 360 degrees wrap (9.8 and 6.8 per cent; both P < 0.001 versus 90 degrees and 180 degrees groups). CONCLUSION: Tailoring the degree of fundoplication according to preoperative oesophageal motility by standard manometric parameters has no long-term impact on postoperative dysphagia. There is, however, a proportionate increase in short term dysphagia scores with increasing degree of wrap, and a corresponding proportionate increase in dilatations and reoperations for dysphagia. These differences in dysphagia scores diminish with time. PMID- 21647869 TI - The CAPRA-S score: A straightforward tool for improved prediction of outcomes after radical prostatectomy. AB - BACKGROUND: The authors previously developed and validated the Cancer of the Prostate Risk Assessment (CAPRA) score to predict prostate cancer recurrence based on pretreatment clinical data. They aimed to develop a similar postsurgical score with improved accuracy via incorporation of pathologic data. METHODS: A total of 3837 prostatectomy patients in the Cancer of the Prostate Strategic Urologic Research Endeavor (CaPSURETM) national disease registry were analyzed. Cox regression was used to determine the predictive power of preoperative prostate-specific antigen (PSA), pathologic Gleason score (pGS), surgical margins (SM), extracapsular extension (ECE), seminal vesicle invasion (SVI), and lymph node invasion (LNI). Points were assigned based on the relative weights of these variables in predicting recurrence. The new postsurgical score (CAPRA-S) was tested and compared with a commonly cited nomogram with proportional hazards analysis, concordance (c) index, calibration plots, and decision-curve analysis. RESULTS: Recurrence appeared in 16.8% of the men; actuarial progression-free probability at 5 years was 78.0%. The CAPRA-S was determined by adding up to 3 points for PSA, up to 3 points for pGS, 1 point each for ECE and LNI, and 2 points each for SM and SVI. The hazard ratio for each point increase in CAPRA-S score was 1.54 (95% confidence interval, 1.49-1.59), indicating a 2.4-fold increase in risk for each 2-point increase in score. The CAPRA-S c-index was 0.77, substantially higher than 0.66 for the pretreatment CAPRA score and comparable to 0.76 for the nomogram. The CAPRA-S score performed better in both calibration and decision curve analyses. CONCLUSIONS: The CAPRA-S offers good discriminatory accuracy, calibration, and ease of calculation for clinical and research settings. PMID- 21647870 TI - Making and evaluating a statistical prediction model for the absolute risk of prostate cancer recurrence. PMID- 21647872 TI - Single amino acid arginine starvation efficiently sensitizes cancer cells to canavanine treatment and irradiation. AB - Single amino acid arginine deprivation is a promising strategy in modern metabolic anticancer therapy. Its potency to inhibit tumor growth warrants the search for rational chemo- and radio-therapeutic approaches to be co-applied. In this report, we evaluated, for the first time, the efficacy of arginine deprivation as anticancer therapy in three-dimensional (3D) cultures of human tumor cells, and propose a new combinatorial metabolic-chemo-radio-treatment regime based on arginine starvation, low doses of arginine natural analog canavanine and irradiation. A sophisticated experimental setup was designed to evaluate the impact of arginine starvation on four human epithelial cancer cell lines in 2D monolayer and 3D spheroid culture. Radioresponse was assessed in colony formation assays and by monitoring spheroid regrowth probability following single dose irradiation using a standardized spheroid-based test platform. Surviving fraction at 2 Gy (SF(2Gy)) and spheroid control dose(50) (SCD(50) ) were calculated as analytical endpoints. Cancer cells in spheroids are much more resistant to arginine starvation than in 2D culture. Spheroid volume stagnated during arginine deprivation, but even after 10 days of starvation, 100% of the spheroids regrew. Combination treatment, however, was remarkably efficient. In particular, pretreatment of cancer cells with the arginine-degrading enzyme arginase combined with or without low concentration of canavanine substantially enhanced cell radioresponse reflected by a loss in spheroid regrowth probability and SCD(50) values reduced by a factor of 1.5-3. Our data strongly suggest that arginine withdrawal alone or in combination with canavanine is a promising antitumor strategy with potential to enhance cancer cure by irradiation. PMID- 21647871 TI - A phase 1b trial of the combination of the antiangiogenic agent sunitinib and radiation therapy for patients with primary and metastatic central nervous system malignancies. AB - BACKGROUND: In this phase 1 trial, the authors evaluated sunitinib combined with radiation therapy (RT) for the treatment of primary or metastatic central nervous system (CNS) malignancies. METHODS: Eligible patients had CNS malignancies that required a (minimum) 2-week course of RT. Sunitinib (37.5 mg) was administered daily for the duration of RT with optional treatment extension of 1 month. Urine was collected at 3 time points for correlative biomarker studies. The primary endpoint was acute toxicity defined according to Common Toxicity Criteria version 3. RESULTS: Fifteen patients were enrolled (12 with CNS metastasis and 3 with primary tumors). RT doses ranged from 14 Gray (Gy) to 70 Gy (1.8-3.5 Gy per fraction). Acute toxicities included hematologic, nausea, hyperglycemia, fatigue, hypocalcemia, and diarrhea. Six patients (40%) developed grade <= 2 toxicities. Grade 3 toxicities occurred in 7 patients (47%) and included hematologic toxicity, fatigue, deep vein thrombosis, dysphasia, hyperglycemia, and hyponatremia. No grade 3 through 5 hypertensive events or intracerebral hemorrhages occurred. Two grade 5 adverse events attributed to disease progression occurred. The median follow-up was 34.2 months. Two patients (13%) achieved a partial response, 9 patients (60%) had stable disease, and 2 patients (13%) patients had progressive disease. The 6-month progression-free survival rate for patients who had brain metastasis was 58%. Grade 3 hematologic toxicity was correlated with greater changes in vascular endothelial growth factor levels changes between baseline and the completion of RT. CONCLUSIONS: Continuous 37.5 mg sunitinib combined with RT in patients who had CNS malignancies yielded acceptable toxicities and adverse events. The current results indicated that changes in urine vascular endothelial growth factor levels are associated with hematologic toxicity, and this association should be analyzed in a larger cohort. The feasibility, safety, and early response results warrant a phase 2 trial. PMID- 21647873 TI - MDM2 promoter SNP309 is associated with an increased susceptibility to chronic lymphocytic leukemia and correlates with MDM2 mRNA expression in Chinese patients with CLL. AB - A single nucleotide polymorphism (SNP) at position 309 in the promoter region of MDM2 leading to increased expression of MDM2 and attenuated function of p53 has recently been suggested as an unfavorable prognostic marker in chronic lymphocytic leukemia (CLL) although this has been questioned. The MDM2 SNP309 genotypes in 173 CLL patients and 260 healthy controls were detected by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method, which was confirmed by direct DNA sequencing. Compared with the T/T genotype, the SNP309 G/G genotype instead of T/G heterozygote was associated with a significantly increased risk of CLL (OR = 2.84; 95% CI 1.61-5.03; p < 0.001). Age at onset of CLL was similar irrespective of MDM2 status. MDM2 mRNA expression within CLL of G/G genotype was significantly higher than that in T/G (p = 0.009) and T/T genotypes (p < 0.001). Excluding patients with p53 deletions or mutations enhanced the significance of the findings (G/G vs. T/T, p < 0.001; G/G vs. T/G p = 0.001), which prompted us to study the role of the polymorphism in p53 wild type individuals. In the p53 wild-type groups, survival analysis showed that the patients with MDM2 SNP309 G/G and T/G genotypes both had significantly shorter treatment-free survival (TFS) than SNP309 T/T genotype. Notably, univariate and multivariate analyses showed that MDM2 SNP309 genotypes were associated with TFS. These data show that MDM2 309G polymorphisms contribute to the risk of developing CLL. The unfavorable MDM2 SNP309 G/G genotype was associated with an increase of MDM2 mRNA expression. MDM2 SNP309 was found to be associated with TFS in p53 wild type Chinese CLL populations. PMID- 21647874 TI - alphaL beta2 integrin is indispensable for CD8+ T-cell recruitment in experimental pancreatic and hepatocellular cancer. AB - Recruitment of activated leukocytes from peripheral blood into the tumor tissue is a crucial step of the immune response, which is controlled by the interaction between specific adhesion molecules such as endothelial ICAM-1 and leukocyte beta(2) -integrins. Although attenuated expression of adhesion molecules on tumor endothelium has been proposed to represent a mechanism, which suppresses the intratumoral leukocyte infiltration, the relevance of adhesion molecules for leukocyte recruitment in tumor tissue is poorly understood. The present study is the first investigation of the role of ICAM-1 and beta(2) -integrins in leukocyte recruitment in pancreatic and hepatocellular cancer in vivo, which was studied using knockout mice, intravital time-lapse microscopy and immunohistochemistry. We found that tumor tissue of both pancreatic and hepatocellular cancer was infiltrated with numerous active lymphoid and myeloid leukocytes, although the leukocyte extravasation rate in tumor blood vessels was very low. The knockout of LFA-1 (also known as alpha(L) beta(2) integrin) strongly suppressed recruitment of CD8(+) T cells whereas no significant differences of leukocyte adhesion and infiltration were found in ICAM-1(-/-) and Mac-1(-/-) mice. Analysis of the interstitial leukocyte migration demonstrated that intratumoral leukocytes used haptokinetic type of migration, however, no significant differences of leukocyte migration between any knockout strains were found. We concluded that leukocyte recruitment in pancreatic and hepatocellular cancer is a slow-going process whose dynamics clearly contrasts to a high-speed leukocyte recruitment during acute inflammation. In contrast to acute inflammatory reaction, only LFA-1 controls recruitment of CD8(+) T-cells in both pancreatic and hepatocellular cancer, whereas ICAM-1 and Mac-1 are dispensable. PMID- 21647875 TI - ARFGAP3, an androgen target gene, promotes prostate cancer cell proliferation and migration. AB - ADP ribosylation factor GTPase-activating protein 3 (ARFGAP3) is a GTPase activating protein that associates with the Golgi apparatus and regulates the vesicular trafficking pathway. In the present study, we examined the contribution of ARFGAP3 to prostate cancer cell biology. We showed that ARFGAP3 expression was induced by 100 nM of dihydrotestosterone (DHT) at both the mRNA and protein levels in androgen-sensitive LNCaP cells. We generated stable transfectants of LNCaP cells with FLAG-tagged ARFGAP3 or a control empty vector and showed that ARFGAP3 overexpression promoted cell proliferation and migration compared with control cells. We found that ARFGAP3 interacted with paxillin, a focal adhesion adaptor protein that is important for cell mobility and migration. Small interfering RNA (siRNA)-mediated knockdown of ARFGAP3 showed that ARFGAP3 siRNA markedly reduced LNCaP cell growth. Androgen receptor (AR)-dependent transactivation activity on prostate-specific antigen (PSA) enhancer was synergistically promoted by exogenous ARFGAP3 and paxillin expression, as shown by luciferase assay in LNCaP cells. Thus, our results suggest that ARFGAP3 is a novel androgen-regulated gene that can promote prostate cancer cell proliferation and migration in collaboration with paxillin. PMID- 21647876 TI - Prescribing sunshine: a cross-sectional survey of 500 Australian general practitioners' practices and attitudes about vitamin D. AB - This study aimed to assess the attitudes, practices and knowledge of general practitioners (GPs) with regards to vitamin D. A cross-sectional survey of a random sample of GPs stratified by location of practice (rural/remote or metropolitan) and employment status (full-time or part-time) in New South Wales (NSW), Australia was conducted. Of 500 respondents, 58.1% (95% CI 53.8-62.4) reported that up to 39% of their tested patients showed vitamin D deficiency or insufficiency and a further 37.7% (95% CI 33.5-41.9) of respondents said that over 40% of their patients were vitamin D insufficient. Vitamin D supplementation and advice to receive more natural sunlight were the most common ways vitamin D insufficiency was managed (97.1%; 95% CI 95.6-98.6 and 82%, 95% CI 78.6-85.4, respectively). Some gaps in knowledge were identified. Most respondents (64%; 95% CI 59.8-68.2) believed that a person of average sun sensitivity required 10 min of direct sun exposure during summer in peak UV time and a further 21.6% (95% CI 18.0-25.2) believed that people required 30 min of direct sun. A third of respondents (33.1%; 95% CI 29.0-37.2) advised their patients to use sun protection at all times during winter. In general, the attitude items showed that respondents expressed greater concern about vitamin D deficiency than skin cancer. The results reveal some confusion in general practice regarding vitamin D, sun exposure, sun protection and skin cancer risk. Some of the advice that GPs are offering may needlessly increase their patients' risk for vitamin D insufficiency or skin cancer. PMID- 21647877 TI - Deregulation of Snai2 is associated with metastasis and poor prognosis in tongue squamous cell carcinoma. AB - The members of the Snail superfamily of zinc-finger transcription factors, including Snai1 and Snai2, are involved in essential biological processes, such as epithelial-mesenchymal transition (EMT). Although Snai1 has been investigated in a number of cancers, our knowledge on Snai2 and its role(s) in squamous cell carcinoma of oral tongue (SCCOT) is limited. In this study, we confirmed the previous observation that over-expression of Snai2 is a frequent event in SCCOT. We further demonstrated that Snai2 over-expression is associated with lymph node metastasis in two independent SCCOT patient cohorts (total n = 129). Statistical analysis revealed that Snai2 over-expression was correlated with reduced overall survival. Furthermore, over-expression of Snai2 was correlated with reduced E cadherin expression and enhanced Vimentin expression, suggesting a functional role of Snai2 in EMT. These observations were confirmed in vitro, in which knockdown of Snai2 induced a switch from a mesenchymal-like morphology to an epithelial-like morphology in SCCOT cell lines, and suppressed the cell invasion and migration. In contrast, ectopic transfection of Snai2 led to enhanced cell invasion and migration. Furthermore, Snai2 knockdown attenuated TGFbeta1-induced EMT in SCCOT cell lines. Taken together, these data suggest that Snai2 plays major roles in EMT and the progression of SCCOT and may serve as a therapeutic target for patients at risk of metastasis. PMID- 21647878 TI - Mesenchymal-epithelial transitions: spontaneous and cumulative syntheses of epithelial marker molecules and their assemblies to novel cell junctions connecting human hematopoietic tumor cells to carcinomatoid tissue structures. AB - Using biochemical as well as light- and electron-microscopic immunolocalization methods, in cultures of unicellular human blood tumor cells, we have studied the phenomenon of spontaneous and cumulative syntheses of certain epithelial proteins and glycoproteins and their assemblies to two major kinds of novel cell-cell junctions, adhering junctions (AJs) and junctions based on the epithelial cell adhesion molecule (EpCAM). More than two decades, we have selected and characterized clonal sublines of multipotential hematopoietic K562 cells, which are enriched in newly formed AJs based on cis-clusters of desmoglein Dsg2, in some sublines accompanied by desmocollin Dsc2. Both desmosomal cadherins can be anchored in a submembranous plaque containing plakoglobin and plakophilins Pkp2 and Pkp3, with or without other armadillo proteins and desmoplakin. Also, these cells are often connected by an additional, extended junction system, in which the transmembrane epithelial glycoprotein EpCAM is associated with a cytoplasmic plaque rich in several actin-binding proteins such as afadin, alpha-actinin, ezrin and vinculin. Both kinds of junctions contribute to connections of K562 cells into epithelioid monolayers or even three-dimensional, tissue-like structures, thus markedly changing the cell biological nature and behavior of the resulting tumor subforms (mesenchymal-epithelial transitions). We discuss molecular mechanisms involved in the formation and function of these junctions, also with respect to tumor spread and metastasis, as well as diagnostic and therapeutic consequences. PMID- 21647879 TI - Reactivation of p53 mutants by prima-1 [corrected] in thyroid cancer cells. AB - Most undifferentiated thyroid carcinomas express p53 mutants and thereafter, are very resistant to chemotherapy. p53 reactivation and induction of massive apoptosis (Prima-1) is a compound restoring the tumor-suppressor activity of p53 mutants. We tested the effect of Prima-1 in thyroid cancer cells harboring p53 mutations. Increasing doses of Prima-1 reduced viability of thyroid cancer cells at a variable extent (range 20-80%). Prima-1 up-regulated p53 target genes (p21(WAF1) , BCL2-associated X protein (Bax), and murine double minute 2 (MDM2)), in BC-PAP and Hth-74 cells (expressing D259Y/K286E and K286E p53 mutants) but had no effect in SW1736 (p53 null) and TPC-1 (expressing wild-type p53) thyroid cancer cells. Prima-1 also increased the cytotoxic effects of either doxorubicin or cisplatin in thyroid cancer cells, including the chemo-resistant 8305C, Hth-74 and BC-PAP cells. Moreover, real-time PCR and Western blot indicated that Prima-1 increases the mRNA of thyroid-specific differentiation markers in thyroid cancer cells. Fluorescence-activated cell sorting analysis revealed that Prima-1 effect on thyroid cancer cells occurs via the enhancement of both cell cycle arrest and apoptosis. Small interfering RNA experiments indicated that Prima-1 effect is mediated by p53 mutants but not by the p53 paralog p73. Moreover, in C-643 thyroid cancer cells, forced to ectopically express wild-type p53, Prima-1 prevented the dominant negative effect of double K248Q/K286E p53 mutant. Finally, co-IP experiments indicated that in Hth-74 cells Prima-1 prevents the ability of p53 mutants to sequestrate the p53 paralog TAp73. These in vitro studies imply that p53 mutant reactivation by small compounds may become a novel anticancer therapy in undifferentiated thyroid carcinomas. PMID- 21647880 TI - Pleural malignant mesothelioma epidemic: incidence, modalities of asbestos exposure and occupations involved from the Italian National Register. AB - Due to the large scale use of asbestos (more than 3.5 million tons produced or imported until its definitive banning in 1992), a specific national surveillance system of mesothelioma incident cases is active in Italy, with direct and individual anamnestic etiological investigation. In the period between 1993 and 2004, a case-list of 8,868 pleural MM was recorded by the Italian National Register (ReNaM) and the modalities of exposure to asbestos fibres have been investigated for 6,603 of them. Standardized incidence rates are 3.49 (per 100,000 inhabitants) for men and 1.25 for women, with a wide regional variability. Occupational asbestos exposure was in 69.3% of interviewed subjects (N = 4,577 cases), while 4.4% was due to cohabitation with someone (generally, the husband) occupationally exposed, 4.7% by environmental exposure from living near a contamination source and 1.6% during a leisure activity. In the male group, 81.5% of interviewed subjects exhibit an occupational exposure. In the exposed workers, the median year of first exposure was 1957, and mean latency was 43.7 years. The analysis of exposures by industrial sector focuses on a decreasing trend for those traditionally signaled as "at risk" (asbestos-cement industry, shipbuilding and repair and railway carriages maintenance) and an increasing trend for the building construction sector. The systematic mesothelioma surveillance system is relevant for the prevention of the disease and for supporting an efficient compensation system. The existing experience on all-too-predictable asbestos effects should be transferred to developing countries where asbestos use is spreading. PMID- 21647881 TI - Mindfulness as a moderator in expressive writing. AB - Randomized, controlled studies have documented positive physical and psychological effects of writing about traumatic stress. Some of these studies have shown that individual differences play an important role, with participants responding differently to the intervention based on their personal characteristics. In the present expressive writing experiment, the trait of mindfulness was examined as a potential moderator. Seventy-six undergraduates were randomly assigned to either expressive writing (n = 37) or a control group (n = 39). Main effects favoring expressive writing were found, and these were qualified by significant interactions with mindfulness. Specifically, individuals with higher mindfulness scores responded better to expressive writing, experiencing greater physical and psychological benefits than individuals with lower mindfulness scores. PMID- 21647883 TI - The psychotherapy of psychotherapists. AB - This article introduces a special issue of the Journal of Clinical Psychology: In Session that provides 6 psychologists' narrative accounts of their own personal therapies and a practice-friendly research review on the characteristics of therapist-patients and their own treatment experiences. In response to a standard set of questions, highly experienced psychotherapists hailing from diverse theoretical commitments wrote the accounts. Their accounts illuminate subtle nuances of the therapeutic relationship and treatment outcome, perhaps more fully than other sources of data. Much of value can be learned from these essays and the research findings about the linkages between receiving and conducting psychotherapy and about the technical and emotional challenges that arise when treating a patient who shares the same profession. PMID- 21647884 TI - My experience with psychotherapy, existential analysis and Jungian analysis: Rollo May and beyond. AB - This article describes my initial psychotherapy experience with a psychologist who combined a client-centered/rational-emotive approach, my existential analysis with Rollo May, and then concludes briefly portraying my current Jungian analysis. I explain how I came to each of these experiences, what I learned from them, and the limitations I have recognized in them. I elaborate on the existential analysis with Rollo May, as it marked a major turning point in my life and thinking. I have been able to describe that experience with the benefit of hindsight, which I do not have with the Jungian analysis. In closing, I offer some suggestions about the role of therapy/analysis in the training of the psychotherapist. PMID- 21647882 TI - Pain and emotion: a biopsychosocial review of recent research. AB - OBJECTIVE AND METHOD: Research on emotion and pain has burgeoned. We review the last decade's literature, focusing on links between emotional processes and persistent pain. RESULTS: Neurobiological research documents the neural processes that distinguish affective from sensory pain dimensions, link emotion and pain, and generate central nervous system pain sensitization. Psychological research demonstrates that greater pain is related to emotional stress and limited emotional awareness, expression, and processing. Social research shows the potential importance of emotional communication, empathy, attachment, and rejection. CONCLUSIONS: Emotions are integral to the conceptualization, assessment, and treatment of persistent pain. Research should clarify when to eliminate or attenuate negative emotions, and when to access, experience, and express them. Theory and practice should integrate emotion into cognitive behavioral models of persistent pain. PMID- 21647885 TI - Lessons from the deep study of rare tumours. AB - Next-generation sequencing technologies are having an enormous impact on mapping mutations in cancer. However, it is unclear to what extent mutations in genes are shared between cancer types, and to what extent these are unique to different cancers. While the mapping of mutations is almost saturated in common cancer types, the study of rare tumours offers surprising insights into pathways that are deranged in cancer. The paper by Amary et al. in this issue of the Journal of Pathology illustrates the value of studying uncommon cancer types. The authors report on a startlingly high incidence of IDH1/2 mutations in cartilaginous tumours. This finding not only represent a major step forward in mapping the molecular pathogenesis of these tumours, but provides further evidence of the intriguing roles of metabolic pathways in carcinogenesis. PMID- 21647887 TI - Interaction between Eudragit(r) E100 and anionic drugs: addition of anionic polyelectrolytes and their influence on drug release performance. AB - In this work, we report results concerning the study of solid complexes compounded by a cationic polymethacrylate (Eudragit(r) E100, Eu) and mesalazine (M) (Eu-M(x) complex). The influence of an anionic polyacrylic acid polymer (carbomer, C) on dissolution behavior of M from the complex was evaluated (Eu M(x) C(y) complex). The dissolution profiles and solvent front movements of solid matrices in different media (water, buffer pH 7.4, 0.9% NaCl) were investigated and ionic interactions among Eu, M, and C were determined through Fourier transform infrared (FT-IR) spectroscopy. For Eu-M(x) complexes, the affinity between M and Eu modulated the delivery of free M in solution, with the dissolution media affecting the delivery rate mainly due to an ionic interchange process between M and anionic electrolytes (i.e., Cl(-)). FTIR spectroscopy allowed the ionic interaction between Eu and M to be verified. The addition of C (Eu-M(x) C(y) ) influenced the dissolution behavior of these matrices. As the amount of C was increased, the release mechanism changed from diffusion (Eu-M(50) ) or anomalous (Eu-M(100)) to zero order (Eu-M(x) C(50)). This variation in rate delivery was also affected by the dissolution media, as occurred with Eu-M(x) complexes. The formation of the gel layer during the dissolution process, as consequence of Eu-M(x) C(y) matrices hydration, was influenced by C amount and dissolution media. PMID- 21647886 TI - Multidimensional methods for the formulation of biopharmaceuticals and vaccines. AB - Determining and preserving the higher order structural integrity and conformational stability of proteins, plasmid DNA, and macromolecular complexes such as viruses, virus-like particles, and adjuvanted antigens are often a significant barrier to the successful stabilization and formulation of biopharmaceutical drugs and vaccines. These properties typically must be investigated with multiple lower resolution experimental methods because each technique monitors only a narrow aspect of the overall conformational state of a macromolecular system. This review describes the use of empirical phase diagrams (EPDs) to combine large amounts of data from multiple high-throughput instruments and construct a map of a target macromolecule's physical state as a function of temperature, solvent conditions, and other stress variables. We present a tutorial on the mathematical methodology, an overview of some of the experimental methods typically used, and examples of some of the previous major formulation applications. We also explore novel applications of EPDs including potential new mathematical approaches as well as possible new biopharmaceutical applications such as analytical comparability, chemical stability, and protein dynamics. PMID- 21647888 TI - Analytical characterization of a novel degradation product in a PEGylated recombinant protein. AB - We report the identification and characterization of a novel degradation product associated with PEGylation of a recombinant protein. After several months of storage at 2 degrees C-8 degrees C, an unexpected increase was observed in the proportion of an impurity that eluted with the native unPEGylated protein by size exclusion chromatography--from less than 0.01% at the start of storage to more than 0.25% at 12 months. An investigation into the nature of the impurity determined the presence of an N-terminal adduction with a mass increase of +58 Da over the native unPEGylated protein species, demonstrating that this impurity was the result of degradation. The impurity was subjected to thorough analytical characterization using orthogonal methods to establish its identity, and a mechanistic model proposed for its formation. The data implicate the presence of a monomethoxy polyethylene glycol (mPEG)-acetal aldehyde impurity in the mPEG aldehyde raw material, indicating the need for diligent raw material testing prior to use. PMID- 21647889 TI - Cochlear reimplantation with same device: surgical and audiologic results. AB - OBJECTIVES/HYPOTHESIS: To study to what extent it is possible to achieve identical insertion depths and to maintain the same performance after cochlear reimplantation. STUDY DESIGN: Outcome research on a retrospective case series in a tertiary university referral center. METHODS: Data were collected for 12 adults and three children who underwent reimplantation during the last 3 years with a new HiRes90K device with HiFocus 1J electrode owing to failure of the feed through seal. Multislice computed tomography scans were used to compare positions of the original and newly placed electrode arrays. The speech-perception scores on a consonant-vowel-consonant word test before and after reimplantation were compared. RESULTS: All reimplantations were successfully performed by two experienced cochlear implantation surgeons, and no complications were observed. Postoperative imaging showed that the average displacement of the new implant was only 0.59 mm. Reactivation of the implant gave immediate open set speech understanding in all patients, and speech perception rapidly returned to the previous level obtained with the original implant within weeks; it was even significantly better at the 3-month follow-up. No relation was found between changes in performance and the amount of displacement of the electrode array. CONCLUSIONS: After cochlear reimplantation with the same device, electrode-array position can be accurately replicated and speech perception can be regained or even improved within weeks. PMID- 21647890 TI - Perioperative incidence and management of hyponatremia in vWD patients undergoing adenotonsillectomy. AB - OBJECTIVES/HYPOTHESIS: To analyze the incidence and severity of hyponatremia in patients receiving synthetic desmopressin (DDAVP) in the perioperative setting of oropharyngeal surgery in the treatment of von Willebrand disease and to propose a standardized protocol for perioperative fluid resuscitation and postoperative sodium monitoring after DDAVP administration. STUDY DESIGN: Retrospective medical record review. METHODS: A retrospective medical record review in an academic pediatric medical center was conducted. From October 1, 2002, to February 1, 2009, all patients undergoing adenotonsillectomy and receiving DDAVP preoperatively for the treatment of von Willebrand disease were identified. A total of 76 patients were identified by initial database review; 63 patients were included in the study, and 13 patients were excluded secondary to incomplete data. DDAVP dose and timing, perioperative fluid volume and composition, and postoperative sodium levels were collected. Extreme adverse events related to hyponatremia were recorded. RESULTS: Forty-seven of 63 (74.6%) patients developed some degree of hyponatremia after DDAVP administration, and six of 63 (9.5%) patients developed extreme hyponatremia, with the degree of hyponatremia related to the volume of perioperative fluid resuscitation. The sodium nadir occurred within 9 to 20 hours after DDAVP administration. No serious adverse events related to hyponatremia were recorded during the study period. CONCLUSIONS: The incidence of hyponatremia in children receiving DDAVP for prophylaxis of intraoperative bleeding following oropharyngeal surgery is high. The degree of hyponatremia is related to the perioperative fluid volume administered. A protocol for DDAVP administration, perioperative fluid resuscitation, and postoperative sodium monitoring that aims to reduce the incidence of hyponatremia in this population is proposed. PMID- 21647891 TI - The effects of verbal reaction time in Alzheimer's disease. AB - OBJECTIVES/HYPOTHESIS: Verbal fluency deteriorates with normal aging, but is much more severe in Alzheimer's Disease (AD). Verbal functions were analyzed to find differences between normal aging subjects in patients with mild cognitive impairment (MCI), and in patients with early and moderate stages of AD. This study measured the verbal response time in patients with AD, MCI, and in control subjects STUDY DESIGN: This study measured the verbal response time in patients with AD, MCI, and in control subjects METHODS: Fifteen patients with MCI, 15 patients with early AD, 8 patients with moderate AD, and 15 subjects for controls were included in the study. Word length in milliseconds, reaction time to a phoneme, word, or sentence and acoustic analysis of voice quality and speech diadochokinetic rate (DDK) were measured. RESULTS: Reaction time for a phoneme, word, or sentence especially the initiation period for them were longer in patients with early AD compared to patients with MCI (P < .001). The mean DDK rate was lower with increased severity of the disease, and was much more severe in patients with moderate AD. CONCLUSIONS: Clinical discrimination of the early stages of AD and MCI is challenging. Unfortunately, there are no laboratory markers present for the diagnosis of preclinical cases of AD. With the results of this study, the assessments of verbal reaction time may helpful for diagnosis of early AD. PMID- 21647892 TI - Prevention of buccal mucosa scarring with transforming growth factor beta3. AB - OBJECTIVES/HYPOTHESIS: In the treatment of tumorous diseases, scarring often forms after resection or irradiation. Scarring of the buccal mucosa causes difficulty in opening the mouth and mastication, decreasing quality of life. Transforming growth factor (TGF) beta3 is an isoform of TGFbeta1 that is known to accelerate scarring, although it has different effects on wound healing. TGFbeta3 administration into wounds has been associated with improvement in the quality of healing skin in vivo. TGFbeta3 is also considered to be an important anti scarring factor in buccal mucosa. The present study aimed to examine whether TGFbeta3 is effective for prevention and treatment of buccal mucosa scarring. STUDY DESIGN: Prospective study using an animal model. METHODS: Thirty Sprague Dawley rats were involved in this study. We injected 0.5 mL of TGFbeta3 (0.005 MUg/mL, 0.05 MUg/mL, 0.5 MUg/mL, 5 MUg/mL) or saline was injected into the buccal submucosa. Fifteen minutes after the injection, the mucosa was removed down to the masseter muscle or orbicularis oris muscle layer using a 6-mm biopsy punch. Six weeks after the operation, the buccal mucosae were harvested after euthanasia. Morphologic and histologic examinations were performed. RESULTS: The administration of 0.5 MUg/mL TGFbeta3 induced rapid re-epithelialization and suppressed scar formation. In the submucosal layer, favorable restoration of hyaluronic acid and elastin was seen in the TGFbeta3-treated groups compared to the saline-treated group. CONCLUSIONS: TGFbeta3 is considered to be effective for better restoration of extracellular matrices of injured buccal mucosa, suggesting a preventative effect of buccal mucosa scarring. PMID- 21647893 TI - Follow-up after gamma knife radiosurgery for vestibular schwannomas: volumetric and axial control rates. AB - OBJECTIVES/HYPOTHESIS: A prospective long-term follow-up study was conducted to evaluate the results of gamma knife radiosurgery (GKRS) for vestibular schwannoma (VS) patients. Both axial and volumetric measurements are used to determine tumor size during follow-up. STUDY DESIGN: Individual prospective cohort study. METHODS: A total of 110 VS patients were referred for radiosurgery between 2002 and 2007. All patients were treated with a Leksell 4C gamma knife. There were 12.5 to 13 Gy prescribed to the isodose covering 90% of the tumor volume. The resulting marginal dose was on average 11.0 Gy (range, 9.3-12.5 Gy). Tumor size and tumor volume were determined before and after gamma knife treatment at regular intervals. The minimal follow-up period was 2 years. RESULTS: There were 100 patients included in the study. Eight patients needed additional treatment after a mean follow-up period of 38 months. One patient experienced a temporary facial nerve deficit. No growth pattern could be recognized for tumor growth after GKRS. Based on the measurements of the largest extrameatal diameter, the tumor size would have decreased or remained stable in 94%. Based on volumetric measurement, the tumor size was decreased or remained stable in 79%. CONCLUSIONS: High tumor control and low complication rates make GKRS a good therapy for VS. If tumor growth occurs after GKRS, a conservative management can be considered because continued tumor growth is uncertain. The extrameatal diameter on axial magnetic resonance imaging seems to be a reliable parameter of the size of a VS. Volumetry is the preferred method to assess the dimensions of a VS, although the consequences of strong volumetric increase, especially in small tumors, can be different depending on individual differences in tumor size. PMID- 21647894 TI - Robotic-assisted lingual tonsillectomy. PMID- 21647895 TI - Dual immunofluorescence staining of proteoglycans in human temporal bones. AB - OBJECTIVES/HYPOTHESIS: Immunofluorescence staining methods have been developed to study the distribution of macromolecules in archival formalin-fixed celloidin embedded human temporal bone tissues. The aim of this study was to investigate the feasibility of utilizing this approach to evaluate the codistribution of more than one molecule of interest in a single tissue section. STUDY DESIGN: Retrospective study of proteoglycan codistribution in archival human temporal bone tissues. METHODS: The chondroitin sulfate and keratan sulfate proteoglycans were selected for evaluating this methodology. Human tissues with known proteoglycan staining patterns were studied as controls. Thirty-one formalin fixed celloidin-embedded archival human temporal bones were evaluated, and the observations in 11 specimens are described. A dual immunofluorescence staining method was developed using primary antibodies of differing isotypes and secondary antibodies labeled with fluorophores having nonoverlapping emission characteristics. RESULTS: The specificity of the dual immunofluorescence technique for chondroitin sulfate and keratan sulfate proteoglycans was demonstrated in control tissues and confirmed through inhibition studies. The normal human tectorial membrane exhibited intense chondroitin sulfate staining. Cochlear and vestibular hair cells exhibited predominantly keratan sulfate staining. Keratan sulfate staining predominated in spiral ganglion cell bodies and fibers. Alterations in the normal distribution pattern of proteoglycans were observed in cases of presbycusis and otosclerosis. CONCLUSIONS: The dual immunofluorescence staining methodology can be used to study archival formalin fixed celloidin-embedded human temporal bone tissues. This technique may be applied to the evaluation of other molecules in archival human temporal bone tissues and lead to improvement in our understanding of the function of these molecules and their role in disease processes. PMID- 21647896 TI - In reference to An electron microscopic study--correlation of gastroesophageal reflux disease and laryngopharyngeal reflux. PMID- 21647897 TI - MRI scanning in patients implanted with a Vibrant Soundbridge. AB - OBJECTIVE: To observe the in vivo effects of MRI scanning on the Vibrant Soundbridge system. STUDY DESIGN: Retrospective questionnaire. METHOD: Sixty three implantees answered a retrospective questionnaire covering their medical/otological and physical conditions pre-, intra-, and post-magnetic resonance imaging scanning (MRI). Bone conduction (BC) thresholds were measured after MRI scanning and compared with the prescan BC. RESULTS: Thirteen implantees (20.6%) underwent 19 MRI scans (1; 1.5 T) for different medical indications (e.g., exclusion of a brain tumour, lumbar disc herniation etc.). Scanner-related impulse noise, pain in the middle ear, or pressure at the receiver bed, as well as changes of the transfer function of the floating mass transducer (FMT) are observed frequent effects of MRI scanning. Two patients required transtympanal repositioning of the FMT. A subjectively reported or objectively documented sensorineural hearing loss was not found in any of our patients in this series. CONCLUSION: MRI scanning with an implanted Vibrant Soundbridge has possible major side effects, but did not affect cochlear function in this series. PMID- 21647899 TI - Oncological results after surgical treatment of squamous cell cancer of the lateral wall of the oropharynx. AB - OBJECTIVES: The gold standard of treatment of cancer of the lateral wall of the oropharynx continues to be unclear, especially in advanced stages. In this study, we report our experience with surgical treatment of these cancers and describe the functional and oncological results of the procedures. STUDY DESIGN: Retrospective review. METHODS: A total of 155 previously untreated patients with squamous cell carcinoma of the lateral wall of the oropharynx who underwent a surgical resection of the lesion at our department from January 1990 to January 2008 were included. Sixty-seven percent of these patients received postoperative radiotherapy. The records of these patients were reviewed to obtain measures such as local and regional control, disease-specific survival, and speech and swallowing function. RESULTS: Six patients had a stage I disease, 15 had a stage II disease, 31 had a stage III disease, 86 had a stage IVA, and 17 had stage IVB disease. The overall recurrence rate was 60%, and the local recurrence rate was 40%. The 5-year overall survival and disease-specific survival rates were 33% and 43%, respectively. Five-year disease-specific survival rates by stage were as follows: 100%, 59%, 57%, 31%, and 33% for stages I to IVB, respectively. Multivariate analysis showed two parameters that were independent predictors of a reduced disease-specific survival: cervical lymph node metastases pN2-3 (P = .027) and primary tumor classified as pT3-4 (P = .029). In 122 patients, a tracheotomy was performed, and it couldn't be sealed in 23% of them. Oral alimentation was successfully recovered in 93% of the patients. CONCLUSIONS: Surgical treatment of cancer of the lateral wall of the oropharynx provides acceptable oncological and functional results, especially in early and moderately advanced stages (stages I-III). In advanced stages (stage IV), we obtained good functional preservation rates but poor oncological outcomes. Consequently, these groups of patients could be considered for another treatment modality, such as radiochemotherapy. PMID- 21647900 TI - Four-handed, two-surgeon microsurgery in neurotology. AB - INTRODUCTION: Since the early 1960s, microsurgical removal has been the standard for curative treatment of vestibular schwannomas and neurotologic skull base lesions. The narrow operative corridor with only one surgeon actively operating sometimes interferes with simultaneous application of standard surgical principles such as suction, counter tension, and dissection and makes attempts at "co-surgery" into "sequential" surgery. Surgical efficiency is often delayed by the need to change surgical instruments and reorient the surgical field. METHODS: In this four-handed technique, the microscope is arranged with binocular microscope heads and the surgeons facing each other at the head of the patient and operating simultaneously. RESULTS: Since 2006, the authors have operated 97 lesions using the four-handed technique. The distinct visualization advantage of the 180 degrees binocular arrangement over use of a side arm is that both surgeons have the same stereoscopic perspective. Specific ergonomic advantages were identified for rapid debulking in large tumors, simultaneous neurologic mapping for sharp facial nerve dissection, counter traction for tumor dissection, neurologic tissue retraction/protection for safe bipolar cautery, and real-time intraoperative consultation/collaboration. CONCLUSIONS: Specific technical and microscope arrangements are necessary to utilize the four-handed technique; however, distinct advantages in tissue handling and surgical efficiency are facilitated. PMID- 21647901 TI - A model for training and evaluation of myringotomy and tube placement skills. AB - OBJECTIVES/HYPOTHESIS: Simulation is emerging as a mandatory component of surgical training and a means of demonstrating surgical competency. We designed a cost-effective, low-fidelity model to further acquisition of technical skills related to myringotomy and ventilation tube insertion (M&T). The purpose of the study was to examine the skills trainer as a method of assessment to evaluate competency, timeliness, and procedure confidence in junior residents. STUDY DESIGN: Prospective, randomized. METHODS: A simplistic M&T skills box was developed. General surgery interns (n = 20) with no prior procedure training were randomized to receive either didactic instruction or skills training using the model. One hour of lecture or technical skills training was provided to each group. A blinded examiner evaluated the subjects in both groups before and after training. Performance was measured using a global rating scale, task-specific checklist, and time-to-completion. Pre- and postsession questionnaires assessed procedure confidence. RESULTS: Analysis revealed a trend toward improvement in global rating scores between groups. There was a statistically significant difference in time improvement between groups (P = .0211). The skills lab group felt they could perform the procedure faster and with improved abilities, as compared to the didactic group (P = .0069 and 0.0007, respectively). CONCLUSIONS: Junior surgical residents performed an M&T procedure using a novel, low-cost model. This study demonstrated the skills lab's positive effect on training as measured by global rating scale, time-to-completion, and overall resident confidence. We anticipate its application to be valuable not only in training residents but also in assessing competency. PMID- 21647903 TI - Nasal tip projection and facial attractiveness. AB - OBJECTIVES/HYPOTHESIS: Six nasal tip projection (NTP) ratios from Goode, Simons, Baum, Powell, and Crumley guide clinical and academic practice on quantifying NTP, but none have been empirically correlated with facial attractiveness. This study's objectives were to determine: 1) if there is a correlation between these ratios and facial attractiveness; and 2) which of the six ratios has the greatest linkage to overall facial attractiveness. STUDY DESIGN: Basic research study. METHODS: There were 300 digital portraits of women (ages 18-25 years) randomly paired and morphed to create 300 synthetic lateral facial images rated by 78 raters in the community. NTP ratios were measured in each portrait. RESULTS: None of the ratios correlated with facial attractiveness. For the Baum, Powell, and Simons ratio, facial attractiveness increased as NTP deviated 1 and 2 standard deviations from the ideal, whereas facial attractiveness decreased as NTP deviated from the Goode and Crumley ideal ratios. The most attractive faces had NTP ratios consistent with previous expert opinion findings. CONCLUSIONS: To our knowledge, this is the first study to empirically correlate these six landmark NTP ratios with facial attractiveness. Although there was no correlation with any of the six ratios, the ideal ratios proposed by Goode and Crumley impacted facial aesthetics the most. Although the ideal ratios are useful in establishing rhinoplasty guidelines, they should only be used as a part of the management in achieving an aesthetic face on the whole, as they may not be robust enough to correlate with overall facial attractiveness. PMID- 21647904 TI - Staphylococcus aureus biofilms: Nemesis of endoscopic sinus surgery. AB - OBJECTIVES/HYPOTHESIS: Chronic rhinosinusitis (CRS) patients with biofilms have persistent postoperative symptoms, ongoing mucosal inflammation, and recurrent infections. Recent evidence suggests that biofilms of differing species confer varying disease profiles in CRS patients. We aimed to prospectively investigate the effects of Staphylococcus aureus, Pseudomonas aeruginosa, Haemophilus influenzae, and fungal biofilms on outcomes following endoscopic sinus surgery (ESS). STUDY DESIGN: Prospective blinded study. METHODS: In this prospective blinded study, 39 patients undergoing ESS for CRS assessed their symptoms preoperatively using internationally accepted standardized symptom scoring systems and quality-of-life measures (10-point visual analog scale, Sino-Nasal Outcome Test-20, global severity of CRS). Their sinonasal mucosa was graded (Lund Kennedy scale) and extent of radiologic disease on computed tomography scans scored (Lund-McKay scale). Random sinonasal tissue samples were assessed for different bacterial species forming biofilms by using fluorescent in-situ hybridization and confocal laser microscopy. For 12 months after surgery, CRS symptoms, quality of life, and objective evidence of persisting disease were assessed by using the preoperative tools. RESULTS: Different bacterial species combinations were found in 30 of 39 patients; 60% of these 30 biofilms were polymicrobial biofilms and 70% had S aureus biofilms. Preoperative nasendoscopy and radiologic disease severity were significantly worse in patients with multiple biofilms (P = .02 and P = .01, respectively), and they had worse postsurgery mucosal outcomes on endoscopy (P = .01) requiring significantly more postoperative visits (P = .04). Those with S aureus biofilms progressed poorly with their symptom scores and quality-of-life outcomes, with significant differences in nasendoscopy scores (P = .007). CONCLUSIONS: S. aureus biofilms play a dominant role in negatively affecting outcomes of ESS with persisting postoperative symptoms, ongoing mucosal inflammation, and infections. PMID- 21647905 TI - Postoperative pain after adult tonsillectomy with PlasmaKnife compared to monopolar electrocautery. AB - OBJECTIVES/HYPOTHESIS: Monopolar cautery is the most commonly used technique for tonsillectomy. The aim of the present study is to compare postoperative pain using a new technology, PlasmaKnife tonsillectomy (PKT), in comparison with monopolar cautery tonsillectomy (MCT). STUDY DESIGN: Prospective, randomized, single-blinded, self-controlled study using paired organs. METHODS: Thirty-two adult patients, aged 18 to 30 years, scheduled for tonsillectomy for recurrent tonsillitis were included. Patients were randomly assigned to undergo PKT on one side and MCT on the opposite side. The primary outcome was self-rated daily pain assessed by using a 10-point scale. Patients were provided 21-day pain diaries and were phoned twice weekly by a research assistant to assess pain and remind them to complete diaries. Secondary outcomes included comparisons of operative time, blood loss, and postoperative complications. RESULTS: Repeated measures analysis of variance comparing PKT to MCT during the 21-day postoperative period revealed no difference in postoperative pain between the two groups (P = .131). In addition, total operative time (P = .276) and blood loss (P = .418) did not differ significantly between PKT and MCT. CONCLUSIONS: Adult subjects undergoing PKT do not experience less postoperative pain in comparison to MCT. PMID- 21647906 TI - Outcomes and adverse events of enlarged tracheoesophageal puncture after total laryngectomy. AB - OBJECTIVES/HYPOTHESIS: Enlargement of the tracheoesophageal puncture (TEP) results in aspiration around the voice prosthesis (VP) and may lead to pneumonia. The primary objective was to summarize control of leakage around the VP after conservative management of enlarged TEP. STUDY DESIGN: Retrospective cohort study. METHODS: This 5-year cohort included 194 patients who underwent total laryngectomy (with or without pharyngectomy) and TEP at the University of Texas MD Anderson Cancer Center. Control of leakage around the VP was analyzed at last follow-up after enlarged TEP. Adverse events were compared in patients with and without enlarged TEP. RESULTS: The incidence of enlarged TEP was 18.6% (36 of 194, 95% confidence interval [CI]: 13.0%-24.1%). Conservative methods commonly attempted in lieu of complete TEP closure included placement of an enlarged flange VP (34 of 36, 94%), temporary VP removal (14 of 36, 39%), and TEP-site injection (8 of 36, 22%). At last follow-up, conservative methods controlled leakage around the VP in 81% (29 of 36) of patients. Only two patients required complete TEP closure due to persistent leakage after enlarged TEP. Unresolved leakage was more common in patients with recurrent cancer after laryngectomy (P = .081) and irregular TEP contour (P = .003). Relative to controls without TEP enlargement, patients with enlarged TEP had a three-fold higher risk of pneumonia (relative risk: 3.4, 95% CI: 1.9-6.2) and aspiration of the prosthesis (relative risk: 3.3, 95% CI: 0.8-14.1). CONCLUSIONS: Although the rate of enlarged TEP is relatively low, the complication significantly elevates risk of pneumonia. Prosthetic leakage related to TEP enlargement can often be managed conservatively to avoid complete closure of the TEP. PMID- 21647907 TI - Immunohistochemical detection of pepsin in laryngeal mucosa for diagnosing laryngopharyngeal reflux. AB - OBJECTIVES/HYPOTHESIS: To investigate whether the pepsin immunohistochemical (IHC) staining of the laryngeal mucosa epithelia is an available test for diagnosing laryngopharyngeal reflux (LPR) in clinic. STUDY DESIGN: Prospective case series. METHODS: Biopsy specimens from interarytenoid mucosa of LPR patients (seven acid LPR and eight nonacid LPR) and 21 sex- and age-matched normal controls were obtained for pepsin IHC staining. The diagnosis of LPR was based on 24-hour combined multichannel intraluminal esophageal impedance pH monitoring. The results of IHC staining were semiquantitatively analyzed and scored as negative (-), weakly positive (+), moderately positive (++), and strongly positive (+++). RESULTS: Six of seven acid LPR (85.7%) and six of eight nonacid LPR (75%) mucosa samples were moderate to strongly positive for intracellular pepsin. By contrast, only three of 21 normal controls (14.3%) were moderately positive. The difference in intracellular pepsin between LPR and the normal laryngeal mucosa was statistically significant (P < .01). No significant difference in intracellular pepsin was observed between the acid and nonacid LPR mucosal samples (P = .453). Using weak positivity (+) as a cutpoint, the presence of intracellular pepsin in the laryngeal mucosa had a sensitivity of 100% and a specificity of 47.6% in detecting LPR (P < .05). However, using the moderate positivity (++) as the cutpoint, the pepsin had a slightly decreased sensitivity of 80% but a sharply increased specificity of 85.7% (P < .05) in the detection of LPR. CONCLUSIONS: Pepsin IHC staining of the laryngeal mucosa appears to be a sensitive and specific test for diagnosing LPR in a clinical application. PMID- 21647908 TI - Telemedicine--an efficient and cost-effective approach in parathyroid surgery. AB - OBJECTIVES/HYPOTHESIS: To demonstrate the effectiveness and cost benefit of using telemedicine for the postoperative visit in patients undergoing parathyroidectomy for primary hyperparathyroidism. STUDY DESIGN: Prospective noncontrolled study at a tertiary medical center of a cohort of 39 patients undergoing postoperative care after parathyroidectomy through TeleHealth at a number of sites at various distances from the primary surgical facility. METHODS: From October 2006 through January 2010, 149 patients underwent parathyroidectomy for primary hyperparathyroidism at one tertiary medical center by a single surgeon. Age, sex, distance from the patient's home to the surgical center and to the TeleHealth site, effective completion of the TeleHealth visit, and postoperative complications were recorded. RESULTS: Of the 149 patients who underwent parathyroidectomy, 39 had their postoperative visit using TeleHealth (26%). There were 26 females (67%) and 13 (33%) males. Mean age was 64 years. All visits were effectively carried out and completed with a nurse and the patient at a remote TeleHealth site and the surgeon at the surgical center site. There were no postoperative surgical complications noted with the visits. Average round distance travel saved was 119 miles. The travel distance saved translated into an average savings of $357.00 per patient (which included estimations of transportation costs and lost work time), with further immeasurable benefits to the patient and healthcare system. CONCLUSIONS: TeleHealth is a cost-effective and efficient way to follow-up with patients who have undergone parathyroidectomy, with significant convenience and financial benefits for the patient and healthcare system. PMID- 21647909 TI - CO2 laser ablation and balloon dilation for acquired nasopharyngeal stenosis: a novel technique. AB - OBJECTIVES/HYPOTHESIS: Although acquired nasopharyngeal stenosis (NPS) is frequently attributed to infectious and granulomatous processes, it can also occur secondary to external beam radiation therapy for head and neck cancer. NPS can be treated with local flaps, laser excision, nasal stenting, and combinations thereof. Unfortunately, the postoperative course is frequently complicated by scarring and restenosis that often necessitates multiple revision surgeries. The objective of this study is to report a novel endoscopic approach, employing the use of a flexible CO(2) laser in combination with balloon dilation and mitomycin C application, allowing for the successful treatment of acquired NPS with lasting results. STUDY DESIGN: Retrospective review of case series. METHODS: An endoscope with a working channel is passed transnasally to the location of nasopharyngeal stenosis. A flexible CO(2) laser fiber is then inserted via the working channel of the scope. Precise radial incisions are made on the stenosis using the laser under direct visualization. The laser is removed, and a controlled radial expansion balloon dilation device is inserted, advanced to span the segment of stenosis, and inflated to achieve adequate dilation. Mitomycin-C is then applied topically to the area of dilation. RESULTS: Three patients with severe NPS were treated using this novel technique. All patients had successful long-lasting dilation of NPS without complications over a follow-up period ranging from 12 to 18 months. CONCLUSIONS: Acquired NPS can be successfully treated with durable results with radial laser incisions and controlled radial expansion balloon dilation. PMID- 21647910 TI - Percutaneous dilatational tracheotomy using the tracheotomy endoscope. PMID- 21647911 TI - The effects of unilateral cochlear implantation on the tinnitus handicap inventory and the influence on quality of life. AB - OBJECTIVES/HYPOTHESIS: Cochlear implantation is now the standard of care in patients with significant sensorineural hearing loss. It is well known that patients with severe hearing loss also experience disabling tinnitus. The purpose of this study was to assess the effects of cochlear implants on the perception of tinnitus using the Tinnitus Handicap Inventory (THI). STUDY DESIGN: Prospective, longitudinal study of 142 cochlear implant patients. METHODS: The THI was administered to 142 patients pre- and postimplantation. Outcome measures were obtained 12 months after the implantation. Secondary analyses to examine the correlation between changes in THI scores and outcome measures such as Hearing Handicap Inventory, Hearing in Noise Test (HINT), and short-form 36 (SF-36) quality-of-life scores were performed. RESULTS: Patients demonstrated statistically significant reduction of the THI scores including its subscales (P < .001). Prior to implantation, 37% of patients described their tinnitus as moderate to severe. Postoperatively, this percentage decreased to 10%. Cochlear implantation resulted in complete tinnitus suppression in 37% and tinnitus reduction in another 29% of patients. THI scores significantly correlated with three domains of the SF-36 quality-of-life questionnaire, namely social, emotional, and general health domains. CONCLUSIONS: Cochlear implants have a significant suppressive effect on tinnitus in 66% of implant users. Although the reduction in the subjectively perceived tinnitus was statistically significant, it did not correlate with HINT; however, it did correlate with three quality-of life domains, more significantly for those whose pretreatment conditions were moderate or worse. PMID- 21647912 TI - Intraoperative photodynamic detection of normal parathyroid glands using 5 aminolevulinic acid. AB - OBJECTIVE: It is important to identify and save the normal parathyroid glands during head and neck surgery because of their role in regulating the blood calcium level, yet it is often difficult to localize normal parathyroid glands during surgery. Fluorescence-guided parathyroidectomy in patients with hyperparathyroidism has already proved useful. However, there are few reports of fluorescence-guided localization of normal parathyroid glands in humans. We investigated the utility of fluorescence-guided localization of normal parathyroid glands during thyroidectomy and completed a spectral fluorescence analysis of the accumulation of 5-aminolevulinic acid metabolites in the parathyroid glands. METHODS: Eight patients with benign thyroid disease and five with malignant thyroid tumors were given 20 mg/kg body weight of 5-aminolevulinic acid orally 5 hours before surgery. After the posterior surface of the thyroid gland was exposed and the recurrent laryngeal nerve was identified, we illuminated the area with a violet-blue light of 405 nm. Tissues showing red fluorescence were biopsied to analyze the spectral fluorescence. RESULTS: Under the violet-blue light, normal parathyroid glands showed red fluorescence, while the surrounding structures such as the thyroid gland, muscles, and fat remained nonfluorescent. The spectral peak was observed at 635 nm indicating 5 aminolevulinic acid metabolites. Histopathologically, the biopsied tissue corresponded to normal parathyroid glands. CONCLUSIONS: 5-Aminolevulinic acid is useful to localize the normal parathyroid glands during thyroid surgery in humans. PMID- 21647913 TI - Disruption of ephrin B/Eph B interaction results in abnormal cochlear innervation patterns. AB - OBJECTIVE: To determine the expression patterns of B ephrins and Ephs in the cochlea and identify functional consequences of disruption of ephrin B/Eph B interactions in both cultured spiral ganglion neurons and in the cochlea of live animals. STUDY DESIGN: The expression patterns of various B ephrins and Ephs were determined in mice with Lac-Z mutation. Mice with null function of individual B ephrin and Eph proteins and those with multiple knockouts were studied for cochlear innervation patterns. METHODS: Mice with B ephrins and Ephs disrupted with the beta-galactosidase gene were sacrificed at P6, and their cochleae isolated and processed for Lac-Z staining to determine expression of these proteins in cochlear tissue. Spiral ganglion cells from wild-type as well as ephrin B1 knockout mice were isolated and cocultured with Eph B2 expressing Cos1 cells and neurite lengths were determined. Fluorescent lipophillic dyes were used to label spiral ganglion cell nerve fibers to determine cochlear innervation patterns in wild-type and knockout mice. RESULTS: Eph B1, B2, and ephrin B2 but not B3 was expressed in the cochlea. Eph B2 inhibited outgrowth of spiral ganglion cell axons from wild-type mice, but not from ephrin B1 knockout mice in culture. Knockout mice with null function of ephrin B1 alone or Eph B1, Eph B2, Eph B3 in combination demonstrated abnormal innervation patterns in the organ of Corti. CONCLUSIONS: Disruption of B ephrins and Ephs results in functional consequences in spiral ganglion cells, suggesting that these proteins play a role in establishing normal innervation patterns in the cochlea. PMID- 21647914 TI - Improved growth curve measurements after supraglottoplasty. AB - OBJECTIVES/HYPOTHESIS: To evaluate changes in growth curve measurements after supraglottoplasty and compare these patterns to patients with laryngomalacia treated conservatively. STUDY DESIGN: Retrospective case-control study. METHODS: Children treated surgically for laryngomalacia were compared to control groups of patients who underwent observation or medical treatment only. Body weight the day of surgery and on follow-up visits was plotted on a standardized growth curve and the z-score calculated. RESULTS: A total of 15 patients were included that were observed (group 1), 71 patients that were treated with medical therapy only (group 2), and 29 patients that underwent surgery (group 3). The average z-score at the time of surgery was significantly worse in group 3 (-0.854 +/- 1.080), compared to the z-score at the time of diagnosis in group 1 (-0.086 +/- 0.834), and the time when antireflux therapy was initiated in group 2 (-0.120 +/- 0.979). Within 3 months from surgery, significant improvement on the growth curve was seen in group 3 (P = .009). After 12 months, all three groups approached the mean on the growth curve. CONCLUSIONS: In this series, patients with severe laryngomalacia who warranted surgery presented with significantly lower percentile weight on a standardized growth curve than patients who required only medical or conservative treatment. Substantial improvement in growth curve percentile was seen in the immediate months following supraglottoplasty. Long term follow-up after supraglottoplasty shows that infants return to a mean weight on the growth curve. PMID- 21647915 TI - Transmastoid extradural-intracranial approach for repair of transtemporal meningoencephalocele: a review of 31 consecutive cases. AB - OBJECTIVE: To review the clinical presentation, surgical techniques, and outcomes of the transmastoid extradural-intracranial (TMEDIC) approach for the treatment of transtemporal meningoencephalocele. HYPOTHESIS: The TMEDIC is a safe and effective approach to repair meningoencephalocele originating from the middle or posterior cranial fossa. STUDY DESIGN: Retrospective chart review. SETTING: Academic neurotologic tertiary referral center. PATIENTS: Thirty-one consecutive patients diagnosed with transpetrous meningo(encephalo)cele, with or without cerebrospinal fluid leak, between January of 2003 and October of 2010. INTERVENTION: TMEDIC approach for repairing herniated neural tissue through the tegmen or posterior fossa plate using the combination of autologous cartilage, fascia, and tissue sealant. MAIN OUTCOME MEASURES: Anatomic location, size, and number of defects, presence of herniated brain tissue, pre- and postoperative hearing thresholds, and failure rate. RESULTS: Mean age was 62 +/- 26 years. The etiology was spontaneous in 25/31 (80%), congenital in 3/31 (10%), chronic otitis media in 2/31 (6%), and posttraumatic in 1/31 (4%). Posttympanostomy tube clear otorrhea was the presenting sign in 21/31 (68%) of patients. The mean duration of symptoms was 26 months (range: 1-240). The defect involved the middle fossa (MF) floor in 25/31 (90%). Both the tegmen tympani and mastoideum were involved in 12/31 (39%) of patients and multiple dehiscences were seen in 7/31 (22%). In 17/31 (55%) of cases the size exceeded 1 cm. No recurrences were seen. CONCLUSION: The TMEDIC is a safe and effective method to repair transtemporal meningoencephalocele obviating the need for a middle fossa craniotomy in certain cases. PMID- 21647916 TI - Olfactory outcomes after endoscopic transsphenoidal pituitary surgery. AB - BACKGROUND: Olfaction has been demonstrated to have a great impact on patients' lives. Transsphenoidal endoscopic pituitary surgery is associated with potentially significant damage to olfactory tissues, but to date this issue has been only poorly documented in the literature. STUDY DESIGN: Prospective cohort study comparing olfactory outcomes pre- and postpituitary surgery. METHOD: Patients were administered the University of Pennsylvania Smell Identification Test (UPSIT) preoperatively and again at 6 months postoperatively. The endoscopic transsphenoidal pituitary surgery was carried out using a full middle turbinate preservation protocol. A Hadad-Bassagasteguy (HB) vascularized septal flap was raised in each case. Secondary outcomes included Lund-Kennedy endoscopy scores (LKES) and patient self-report of olfactory disturbance. The results were analysed using a paired t-tests. RESULTS: Seventeen patients met inclusion criteria for the study. Mean preoperative UPSIT value was 37.2 (normosmia), and mean postoperative UPSIT value was 30.8 (moderate hyposmia) (P < .001). All patients were fully healed with normal LKES scores by 6 months. All patients complained of their olfactory dysfunction. CONCLUSIONS: This study is the first to describe postoperative olfactory perturbations suffered by patients undergoing endoscopic transsphenoidal pituitary surgery. We hypothesize that olfactory impairment results from use of the HB flap. We recommend that the possibility of permanent olfactory changes be added to routine patient counseling and consent for this procedure, and that HB flaps be raised judiciously during trannssphenoidal endoscopic procedures. PMID- 21647917 TI - A case of isolated congenital ductus arteriosus aneurysm detected by fetal echocardiography at 38 weeks of gestation. AB - A 30 year-old pregnant woman (G1P0) was diagnosed with fetal ductus arteriosus aneurysm (DAA) at 38 weeks of gestation. The three-vessel view of the heart and the sagittal view of the ductal arch showed a 12-mm fusiform dilatation of the ductus arteriosus. Turbulent flow was detected in it by color Doppler. DAA was confirmed by postnatal echocardiography within 24 hours after birth and it spontaneously closed in the neonatal period. Congenital DAA, usually developed in the third trimester, is potentially fatal due to the possible complications such as spontaneous rupture, dissection, and thromboembolism. PMID- 21647918 TI - Evaluation of the vessels of the cardinal ligament by transrectal ultrasonography with color Doppler imaging. AB - PURPOSE: Radical hysterectomy is the standard treatment for early-stage cervical cancer. Although during surgery the vessels of the cardinal ligament are divided one by one until the splanchnic nerve is identified before they are dissected, there are individual differences in the numbers of vessels. In this preliminary study, we evaluated the accuracy of preoperative detection of the vessels of the cardinal ligament by transrectal sonography with color Doppler imaging (TR-CDUS) with respect to the number of vessels detected intraoperatively. METHODS: We included patients that underwent radical hysterectomy for cervical cancer in our hospital. The vessels of the cardinal ligament were detected preoperatively by TR CDUS. Radical hysterectomy was subsequently performed and the number of vessels found at surgery was recorded. RESULTS: A total of 21 patients were included. The median number of the left vessels detected by TR-CDUS and intraoperatively were 2 (range, 1-4) and 2 (range, 1-5), respectively. The median numbers of the right vessels detected were 3 (range, 2-4) and 3 (range, 2-4), respectively. The accuracy rates were 90.5% for the left cardinal ligament, 85.7% for the right ligament, and 81.0% for both cardinal ligaments. CONCLUSIONS: The results suggest that TR-CDUS may be useful for preoperative detection of the number of the vessels of the cardinal ligament. PMID- 21647919 TI - Contrast-enhanced sonographic diagnosis of unsuspected internal iliac vein thrombosis. AB - We report a rare case of an isolated and unsuspected thrombosis in the right internal iliac vein. The presentation was atypical with regard to the clinical manifestations and the thrombus location. The patient was admitted for an acute abdominal pain in the right iliac fossa. The thrombosis was suspected on the basis of a portal-phase contrast-enhanced abdominal CT but the lack of a delayed phase did not allow confirmation. Contrast-enhanced ultrasound confirmed the diagnosis, by displaying a well-defined filling defect. PMID- 21647920 TI - Cesarean scar pregnancy: a rare cause of uterine arteriovenous malformation. AB - A 38-year-old gravida 4, para 2 woman with a history of two Cesarean sections and one curettage was referred to our hospital, because of painless vaginal bleeding and 6 weeks + 2 days of amenorrhea. The first diagnosis was Cesarean scar pregnancy, managed with methotrexate. Subsequently, an arteriovenous malformation developed, which was diagnosed with color Doppler imaging. The diagnosis was confirmed with angiography. Successful bilateral uterine artery embolization was performed with ethylene vinyl alcohol copolymer (Onyx), n-butyl-2-cyanoacrylate (Histoacryl), and gelfoam. PMID- 21647921 TI - Serine/threonine-protein phosphatase 1 alpha levels are paralleling olfactory memory formation in the CD1 mouse. AB - Although olfactory discrimination has already been studied in several mouse strains, data on protein levels linked to olfactory memory are limited. Wild mouse strains Mus musculus musculus, Mus musculus domesticus and CD1 laboratory outbred mice were tested in a conditioned odor preference task and trained to discriminate between two odors, Rose and Lemon, by pairing one odor with a sugar reward. Six hours following the final test, mice were sacrificed and olfactory bulbs (OB) were taken for gel-based proteomics analyses and immunoblotting. OB proteins were extracted, separated by 2-DE and quantified using specific software (Proteomweaver). Odor-trained mice showed a preference for the previously rewarded odor suggesting that conditioned odor preference occurred. In CD1 mice levels, one out of 482 protein spots was significantly increased in odor-trained mice as compared with the control group; it was in-gel digested by trypsin and chymotrypsin and analyzed by tandem mass spectrometry (nano-ESI-LC-MS/MS). The spot was unambiguously identified as serine/threonine-protein phosphatase PP1 alpha catalytic subunit (PP-1A) and differential levels observed in gel-based proteomic studies were verified by immunoblotting. PP-1A is a key signalling element in synaptic plasticity and memory processes and is herein shown to be paralleling olfactory discrimination representing olfactory memory. PMID- 21647922 TI - Native multimer analysis of plasma and platelet von Willebrand factor compared to denaturing separation: implication for the interpretation of satellite bands. AB - Blue native electrophoresis (BNE) was applied to analyze the von Willebrand factor (vWF) multimers in their native state and to present a methodology to perform blue native electrophoresis on human plasma proteins, which has not been done before. The major difference between this method and the commonly used SDS agarose gel electrophoresis is the lack of satellite bands in the high-resolution native gel. To further analyze this phenomenon, a second dimension was performed under denaturing conditions. Thereby, we obtained a pattern in which each protein sub-unit from the first dimension dissociates into three distinct sub-bands. These bands confirm the triplet structure, which consists of an intermediate band and two satellite bands. By introducing the second dimension, our novel method separates the triplet structure into a higher resolution than the commonly used SDS-agarose gel electrophoresis does. This helps considerably in the classification of ambiguous von Willebrand's disease subtypes. In addition, our method has the additional advantage of being able to resolve the triplet structure of platelet vWF multimers, which has not been identified previously through conventional SDS-agarose electrophoresis multimer analysis. This potential enables us to compare the triplet structure from platelet and plasmatic vWF, and may help to find out whether structural abnormalities concern the vWF molecule in the platelet itself, or whether they are due to the physiological processing of vWF shed into circulation. Owing to its resolution and sensitivity, this native separation technique offers a promising tool for the analysis and detection of von Willebrand disorder, and for the classification of von Willebrand's disease subtypes. PMID- 21647923 TI - Immunoaffinity capillary electrophoresis: a new versatile tool for determining protein biomarkers in inflammatory processes. AB - Many diseases caused by inflammatory processes can progress to a chronic state causing deterioration in the quality of life and a poor prognosis for long-term survival. To address inflammatory diseases effectively, early detection and novel therapeutics are required. However, this can be challenging, in part because of the lack of early predictive biomarkers and the limited availability of adequate technologies capable of the identification/characterization of key predictive biomarkers present in biological materials, especially those found at picomolar concentrations and below. This review highlights the need for state-of-the art methodologies, with high-sensitivity and high-throughput capabilities, for determination of multiple biomarkers. Although many new biomarkers have been discovered recently, existing technology has failed to successfully bring this advancement to the patient's bedside. We present an overview of the various advances available today to extend the discovery of predictive biomarkers of inflammatory diseases; in particular, we review the technology of immunoaffinity capillary electrophoresis (IACE), which combines the use of antibodies as highly selective capture agents with the high resolving power of capillary electrophoresis. This two-dimensional hybrid technology permits the quantification and characterization of several protein biomarkers simultaneously, including subtle structural changes such as variants, isoforms, peptide fragments, and post-translational modifications. Furthermore, the results are rapid, sensitive, can be performed at a relatively low cost, without the introduction of false positive or false negative data. The IACE instrumentation can have relevance to medical, pharmaceutical, environmental, military, cultural heritage (authenticity of art work), forensic science, industrial and research fields, and in particular as a point-of-care biomarker analyzer in translational medicine. PMID- 21647924 TI - Recent developments in capillary and chip electrophoresis of bioparticles: Viruses, organelles, and cells. AB - In appropriate aqueous buffer solutions, biological particles usually exhibit a particular electric surface charge due to exposed charged or chargeable functional groups (amino acid residues, acidic carbohydrate moieties, etc.). Consequently, these bioparticles can migrate in solution under the influence of an electric field allowing separation according to their electrophoretic mobilities or their pI values. Based on these properties, electromigration methods are of eminent interest for the characterization, separation, and detection of such particles. The present review discusses the research papers published between 2008 and 2010 dealing with isoelectric focusing and zone electrophoresis of viruses, organelles and microorganisms (bacteria and yeast cells) in the capillary and the chip format. PMID- 21647925 TI - Concentration and fractionation by isoelectric trapping in a micropreparative scale multicompartmental electrolyzer having orthogonal pH gradients. Part 1. AB - A multicompartmental electrolyzer called ConFrac has been developed and tested for micropreparative-scale isoelectric trapping separations. ConFrac contains n separate, minimalistic isoelectric trapping core units, each with a separate anode compartment, anodic flow-through compartment, collection compartment, cathodic flow-through compartment and a shared cathode compartment. The collection compartments are all isolated from each other and have volumes of 100 MUL each. The liquid held in the collection compartments is stagnant. The respective anodic and cathodic flow-through compartments are hydraulically serially connected to each other by flexible, minimum-length, narrow internal diameter tubes. The respective feed solutions whose volumes are larger and variable are recirculated through the serially connected flow-through compartments. Poly(vinyl alcohol)-based buffering membranes are placed between the anode compartments, anodic flow-through compartments, collection compartments, cathodic flow-through compartments and cathode compartment. The membranes establish two orthogonal pH gradients in ConFrac. The primary pH gradient is parallel with the direction of the recirculating flows and orthogonal to that of the electric field. The secondary pH gradient is parallel with the direction of the electric field and orthogonal to that of the recirculating flows. Since the recirculating liquids are kept in thermostated reservoirs and the residence times in the flow-through compartments are shorter than 2 s, ConFrac can tolerate power loads as high as 2 W without overheating the solutions. The operation and performance of ConFrac has been quantitatively characterized: four 25 MUM ampholytic components were isolated from 5 mL of feed solution in 20 min and their concentration increased approximately 50-fold. PMID- 21647926 TI - Concentration and fractionation by isoelectric trapping in a micropreparative scale multicompartmental electrolyzer having orthogonal pH gradients. Part 2. AB - A micropreparative-scale multicompartmental electrolyzer called ConFrac has been developed and tested for isoelectric trapping separations. ConFrac can be operated in pass-by-pass mode or recirculating mode, using either asymmetrical feeding (feed enters only the anodic or the cathodic flow-through compartment) or symmetrical feeding (feed enters both the anodic and the cathodic flow-through compartment). Symmetrical feeding results in higher processing rates and is the preferred operation mode. Residence time in the flow-through compartments is set as a compromise between processing rate and temperature rise in the effluent. Ampholytic components have been isolated from 5 to 50 mL volumes of micromolar feed solutions and hundredfold concentrated into 100-MUL collection compartments. Samples containing ampholytic analytes in highly conducting salt solutions were readily desalted and fractionated in ConFrac in one operation. pH transients formerly observed in other isoelectric trapping devices were observed in ConFrac as well. The pH transients were caused by the unequal ion transmission rates of the anodic- and cathodic-side buffering membranes. PMID- 21647927 TI - Combining size-exclusion chromatography and fully automated chip-based nanoelectrospray quadrupole time-of-flight tandem mass spectrometry for structural analysis of chondroitin/dermatan sulfate in human decorin. AB - Chondroitin/dermatan sulfate (CS/DS) chain of decorin (DCN) from human skin fibroblasts (HSk) was released by reductive beta-elimination reaction and digested with chondroitin AC I lyase. Enzymatic hydrolysis mixture of CS/DS chains was separated by size-exclusion chromatography (SEC). Collected octasaccharide fraction was subjected to fully automated chip-based nanoelectrospray (nanoESI) quadrupole time-of-flight (QTOF) MS and tandem MS (MS/MS). MS of human skin fibroblasts DCN CS/DS displayed a high complexity due to the large variety of glycoforms, which under chip-nanoESI MS readily ionized to form multiply charged ions. Except for the regularly tetrasulfated octasaccharide, the investigated fraction contained four additional octasaccharides of atypical sulfation status. Two new oversulfated glycoforms and two undersulfated species were identified. Remarkably, the series of decasaccharides discovered in the same SEC pool was found to encompass a trisulfated and a novel hexasulfated [4,5-Delta-GlcAGalNAc(IdoAGalNAc)4] species. MS/MS by collision-induced dissociation (CID) on the [M-4H]4 ion corresponding to the previously not reported [4,5-Delta-GlcAGalNAc(IdoAGalNAc)3](5S) corroborated for a novel motif in which three N-acetylgalactosamine (GalNAc) moieties are monosulfated, 4,5-Delta-GlcA and the first IdoA from the non-reducing end bear one sulfate group each, while the second N-acetylgalactosamine from the reducing end is unsulfated. PMID- 21647928 TI - Indexing molecules with chemical graph identifiers. AB - Fast and robust algorithms for indexing molecules have been historically considered strategic tools for the management and storage of large chemical libraries. This work introduces a modified and further extended version of the molecular equivalence number naming adaptation of the Morgan algorithm (J Chem Inf Comput Sci 2001, 41, 181-185) for the generation of a chemical graph identifier (CGI). This new version corrects for the collisions recognized in the original adaptation and includes the ability to deal with graph canonicalization, ensembles (salts), and isomerism (tautomerism, regioisomerism, optical isomerism, and geometrical isomerism) in a flexible manner. Validation of the current CGI implementation was performed on the open NCI database and the drug-like subset of the ZINC database containing 260,071 and 5,348,089 structures, respectively. The results were compared with those obtained with some of the most widely used indexing codes, such as the CACTVS hash code and the new InChIKey. The analyses emphasize the fact that compound management activities, like duplicate analysis of chemical libraries, are sensitive to the exact definition of compound uniqueness and thus still depend, to a minor extent, on the type and flexibility of the molecular index being used. PMID- 21647929 TI - Combining a polarizable force-field and a coarse-grained polarizable solvent model. II. Accounting for hydrophobic effects. AB - A revised and improved version of our efficient polarizable force-field/coarse grained solvent combined approach (Masella, Borgis, and Cuniasse, J. Comput. Chem. 2008, 29, 1707) is described. The polarizable pseudo-particle solvent model represents the macroscopic solvent polarization by induced dipoles placed on mobile pseudo-particles. In this study, we propose a new formulation of the energy term handling the nonelectrostatic interactions among the pseudo particles. This term is now able to reproduce the energetic and structural response of liquid water due to the presence of a hydrophobic spherical cavity. Accordingly, the parameters of the energy term handling the nonpolar solute/solvent interactions have been refined to reproduce the free-solvation energy of small solutes, based on a standard thermodynamic integration scheme. The reliability of this new approach has been checked for the properties of solvated methane and of the solvated methane dimer, as well as by performing 10 * 20 ns molecular dynamics (MD) trajectories for three solvated proteins. A long time stability of the protein structures along the trajectories is observed. Moreover, our method still provides a measure of the protein solvation thermodynamic at the same accuracy as standard Poisson-Boltzman continuum methods. These results show the relevance of our approach and its applicability to massively coupled MD schemes to accurately and intensively explore solvated macromolecule potential energy surfaces. PMID- 21647930 TI - Gradients for configuration interaction energies with spin-orbit coupling in a semiempirical framework. AB - We present a method for the analytical evaluation of the molecular energy gradient for a semiempirical configuration interaction (CI) wavefunction, taking into account the spin-orbit coupling. We show how to proceed in the simplest case where all the wavefunctions belonging to the CI subspace considered are mixed by the spin-orbit interaction, and we develop an original procedure for the more complex case where only a limited number of CI eigenvectors of the spin-free Hamiltonian are mixed. PMID- 21647931 TI - Theoretical study on the unimolecular decomposition of thiophenol. AB - The potential energy surface for the unimolecular decomposition of thiophenol (C(6)H(5)SH) is mapped out at two theoretical levels; BB1K/GTlarge and QCISD(T)/6 311+G(2d,p)//MP2/6-31G(d,p). Calculated reaction rate constants at the high pressure limit indicate that the major initial channel is the formation of C(6)H(6)S at all temperatures. Above 1000 K, the contribution from direct fission of the S-H bond becomes important. Other decomposition channels, including expulsion of H(2) and H(2)S are of negligible importance. The formation of C(6)H(6)S is predicted to be strong-pressure dependent above 900 K. Further decomposition of C(6)H(6)S produces CS and C(5)H(6). Overall, despite the significant difference in bond dissociation, i.e., 8-9 kcal/mol between the S-H bond in thiophenol and the O-H bond in phenol, H migration at the ortho position in the two molecules represents the most accessible initial channel. PMID- 21647932 TI - Cognitive and physiological responses in humans exposed to a TETRA base station signal in relation to perceived electromagnetic hypersensitivity. AB - Terrestrial Trunked Radio (TETRA) technology ("Airwave") has led to public concern because of its potential interference with electrical activity in the brain. The present study is the first to examine whether acute exposure to a TETRA base station signal has an impact on cognitive functioning and physiological responses. Participants were exposed to a 420 MHz TETRA signal at a power flux density of 10 mW/m(2) as well as sham (no signal) under double-blind conditions. Fifty-one people who reported a perceived sensitivity to electromagnetic fields as well as 132 controls participated in a double-blind provocation study. Forty-eight sensitive and 132 control participants completed all three sessions. Measures of short-term memory, working memory, and attention were administered while physiological responses (blood volume pulse, heart rate, skin conductance) were monitored. After applying exclusion criteria based on task performance for each aforementioned cognitive measure, data were analyzed for 36, 43, and 48 sensitive participants for these respective tasks and, likewise, 107,125, and 129 controls. We observed no differences in cognitive performance between sham and TETRA exposure in either group; physiological response also did not differ between the exposure conditions. These findings are similar to previous double-blind studies with other mobile phone signals (900-2100 MHz), which could not establish any clear evidence that mobile phone signals affect health or cognitive function. PMID- 21647933 TI - Analysis of the effect of mobile phone base station antenna loading on localized SAR and its consequences for measurements. AB - In this work, the effect of antenna element loading on the localized specific absorption rate (SAR) has been analyzed for base station antennas. The analysis was conducted in order to determine whether localized SAR measurements of large multi-element base station antennas can be conducted using standardized procedures and commercially available equipment. More specifically, it was investigated if the antenna shifting measurement procedure, specified in the European base station exposure assessment standard EN 50383, will produce accurate localized SAR results for base station antennas larger than the specified measurement phantom. The obtained results show that SAR accuracy is affected by the presence of lossy material within distances of one wavelength from the tested antennas as a consequence of coupling and redistribution of transmitted power among the antenna elements. It was also found that the existing standardized phantom is not optimal for SAR measurements of large base station antennas. A new methodology is instead proposed based on a larger, box-shaped, whole-body phantom. PMID- 21647934 TI - The expression of Tousled kinases in CaP cell lines and its relation to radiation response and DSB repair. AB - BACKGROUND: The Tousled-like kinases (TLKs) function in processes of chromatin assembly, including replication, transcription, repair, and chromosome segregation. TLK1/1B interacts specifically with the chromatin assembly factor Asf1, a histone H3-H4 chaperone, and with Rad9, a protein involved in DNA repair, and these interactions are believed to be responsible for the action of TLKs in double-strand break repair and radioprotection. METHODS: Western blotting and RT PCR were used to analyze the expression of TLK1, TLK1B, and TLK2 in a panel of prostate cancer (CaP) cell lines. The pattern of radiotolerance in the cell lines was analyzed in parallel. DU145 and PC-3 cells were also probed with assays utilizing transfected plasmids that could be cleaved in vivo with adeno-expressed HO nuclease to assess the potential contribution of TLK1/1B in DSB repair. RESULTS: This is the first report of TLKs' expression in a panel of CaP cell lines and their relationship to radioresistance. Furthermore, expression of TLK1B in non-expressing PC-3 cells rendered them highly resistant to radiation, and conversely, knockdown to TLK1/1B in expressing DU145 reduced their radiotolerance. CONCLUSIONS: TLKs appear to be intimately linked to the pattern of resistance to DNA damage, and specifically DSBs, a finding that was not reported before for any cell lines, and certainly not systematically for human prostate cell lines. PMID- 21647935 TI - Two-sample density-based empirical likelihood tests for incomplete data in application to a pneumonia study. AB - In clinical trials examining the incidence of pneumonia it is a common practice to measure infection via both invasive and non-invasive procedures. In the context of a recently completed randomized trial comparing two treatments the invasive procedure was only utilized in certain scenarios due to the added risk involved, and given that the level of the non-invasive procedure surpassed a given threshold. Hence, what was observed was bivariate data with a pattern of missingness in the invasive variable dependent upon the value of the observed non invasive observation within a given pair. In order to compare two treatments with bivariate observed data exhibiting this pattern of missingness we developed a semi-parametric methodology utilizing the density-based empirical likelihood approach in order to provide a non-parametric approximation to Neyman-Pearson type test statistics. This novel empirical likelihood approach has both a parametric and non-parametric components. The non-parametric component utilizes the observations for the non-missing cases, while the parametric component is utilized to tackle the case where observations are missing with respect to the invasive variable. The method is illustrated through its application to the actual data obtained in the pneumonia study and is shown to be an efficient and practical method. PMID- 21647936 TI - Differential effect of oxidative or excitotoxic stress on the transcriptional profile of amyotrophic lateral sclerosis-linked mutant SOD1 cultured neurons. AB - Amyotrophic lateral sclerosis (ALS) is a progressive, lethal, degenerative disorder of motor neurons. The causes of most cases of ALS are as yet undefined. In a previous study, it was shown that N-methyl-D-aspartate (NMDA) and H(2)O(2) stimuli reduce neuronal survival in cortical neurons in culture (Boutahar et al., 2008). To identify variations in gene expression in response to these neurotoxins in transgenic vs. control cortical neurons cultures, both microarray and RT-PCR analysis were performed. High-density oligonucleotide microarrays showed changes in the expression of about 600 genes involved in protein degradation, neurotrophic factors pathway, cell cycle, inflammation, cytoskeleton, cell adhesion, transcription, or signalling. The most up-regulated genes following H(2)O(2) treatment were involved in cytoskeletal organization and axonal transport, such as ARAP2, KIF17, and DKK2, or in trophic factors pathways, such as insulin-like growth factor-binding protein 4 (IGFBP4), FGF17, and serpin2. The most down-regulated genes were involved in ion transport, such as TRPV1. After NMDA treatment, the most up-regulated genes were involved in protein degradation, such as ubiquitin-conjugating enzyme E2I and cathepsin H, and the most down regulated genes were involved in ion transport, such as SCN7A. We conclude that these neurotoxins act through different transcriptional inductions, and these changes may reflect an adaptative cellular response to the cellular stress induced by the neurotoxins involved in ALS in the presence of mutant human SOD1. PMID- 21647937 TI - Synaptic failure and adenosine triphosphate imbalance induced by amyloid-beta aggregates are prevented by blueberry-enriched polyphenols extract. AB - The potential neuroprotective properties of fruits have been widely recognized. In this study, we evaluated the protective properties of a blueberry extract (BB 4), rich in polyphenols, in a neurodegenerative model induced by amyloid-beta peptide (Abeta). Chronic treatment with Abeta drastically reduced synaptic transmission and the extent of secretory vesicles, which were recovered partially with BB-4. Also, the extract recovered Ca(2+) transients in hippocampal neurons preincubated with Abeta (0.5 and 5 MUM) by about 25% +/- 3% and 30% +/- 2, respectively. In this work, we demonstrate a novel effect of the BB-4 extract on Abeta-induced ATP leakage, in which this extract was able to antagonize the acute ATP leakage but not chronic ATP depletion. On the other hand, BB-4 prevented the uncoupling of mitochondrial function induced by FCCP by about 85%, but it was unable to modify the uncoupling induced by Abeta. The present results strongly indicate that BB-4 plays a role in the process of Abeta aggregation by reducing the toxic species (i.e., 40 kDa). These findings suggest that a blueberry extract can protect neuronal tissue from Abeta toxicity mainly through its antiaggregation property, and its antioxidant properties and mitochondrial membrane potential capacities are secondary mechanisms important in chronic stages. Our work suggests that BB-4 could be an important nutritional complement to neuronal health as well as a potential nutraceutical formulation useful as a dietary supplement in the elderly. PMID- 21647938 TI - ERK1/2 and ERK5 have distinct roles in the regulation of brain-derived neurotrophic factor expression. AB - Neurotrophins play essential roles in the development, differentiation, and survival of neuronal and nonneuronal cells. Alterations in neurotrophin expression have been implicated in a variety of neurodegenerative disorders. Dysregulation of brain-derived neurotrophic factor (BDNF) has been implicated in deficits of long-term potentiation and cognition and may contribute to the development of Alzheimer's disease (AD). In this study, we used complementary pharmacological and molecular approaches to evaluate the role of ERK1/2 and ERK5, two members of the MAPK pathway associated with neuroprotection, in regulating BDNF expression in C6 glial cells and primary astrocytes. Our data revealed that U0126, an inhibitor of both ERK5 and ERK1/2, increased the levels of BDNF mRNA, whereas the MEK1/2-specific inhibitor PD184352 did not, suggesting that ERK5 exerts negative control over BDNF expression. This was supported by experiments in which RNAi-mediated depletion of ERK5 led to an increase in BDNF. In contrast, transfection with constitutively active MEK5 resulted in an inhibition of BDNF expression, confirming the inhibitory role of ERK5 in the regulation of BDNF. Interestingly, transfection with the dominant active mutant of MEK1 (MEKR4F), the upstream activator of ERK1/2, resulted in a modest increase in BDNF levels. Collectively, our data suggest that ERK5 and ERK1/2 exert opposite effects on BDNF expression and support the hypothesis that an imbalance of these two signaling pathways may contribute to the pathology of diseases in which neurotrophin dysregulation is noted. PMID- 21647939 TI - Brain-derived neurotrophic factor modulates antiretroviral-induced mechanical allodynia in the mouse. AB - Nucleoside reverse transcriptase inhibitors (NRTIs) are key components of HIV/AIDS treatment to reduce viral load. However, these drugs can induce chronic neuropathic pain, leading to increased morbidity in HIV patients. This study examines the role of brain-derived neurotrophic factor (BDNF) in the spinal dorsal horn (SDH) in development of mechanical allodynia in male C57BL/6J mice treated with the NRTI stavudine (d4T). After d4T administration, mice developed increased neuronal activity and BDNF expression in the SDH and hind paw mechanical allodynia that was exacerbated by intrathecal BDNF administration. Intrathecal BDNF alone also increased neuronal activity and caused mechanical allodynia. Because excess BDNF amplified d4T-induced mechanical allodynia and neuronal activity, the impact of decreasing BDNF in the SDH was investigated. After d4T, BDNF heterozygous mice were less allodynic than wild-type littermates, which was negated by intrathecal BDNF administration. Finally, pretreatment with intrathecal trkB-Fc chimera prior to d4T or administration of the tyrosine kinase inhibitor K252a 3 days after d4T blocked BDNF-mediated signaling, significantly attenuated the development of mechanical allodynia (trkB-Fc), and decreased neuronal activity (trkB-Fc and K252a). Taken together, these findings provide evidence that BDNF in the SDH contributes to the development of NRTI-induced painful peripheral neuropathy and may represent a new therapeutic opportunity. PMID- 21647940 TI - Synthesis of 4-hydroxycoumarin heteroarylhybrids as potential antimicrobial agents. AB - A new series of 4-hydroxycoumarin derivatives 3a-d was synthesized by the reaction of 3-bromo-4-hydroxy coumarin 1 with various heteroaldehydes 2a-d in good yields. The synthesized compounds were characterized on the basis of their elemental and spectral (IR, (1)H-NMR and mass spectrometry) analysis. All target compounds were evaluated for their in-vitro antimicrobial activity against Streptococcus pyogenes, methicillin-resistant Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumonia, and Escherichia coli bacterial strains and fungal cultures of Candida albicans, Aspergillus fumigatus, Trichophyton mentagrophytes, and Penicillium marneffei by disk diffusion assay with slight modifications. The minimum inhibitory concentration (MIC) was determined for the test compounds as well as for reference standards. Among the tested compounds, 3a has shown the most potent antibacterial as well as antifungal activities. PMID- 21647941 TI - Coordination of DNA replication and recombination activities in the maintenance of genome stability. AB - Across the evolutionary spectrum, living organisms depend on high-fidelity DNA replication and recombination mechanisms to maintain genome stability and thus to avoid mutation and disease. The repair of severe lesions in the DNA such as double-strand breaks or stalled replication forks requires the coordinated activities of both the homologous recombination (HR) and DNA replication machineries. Growing evidence indicates that so-called "accessory proteins" in both systems are essential for the effective coupling of recombination to replication which is necessary to restore genome integrity following severe DNA damage. In this article we review the major processes of homology-directed DNA repair (HDR), including the double Holliday Junction (dHJ), synthesis-dependent strand annealing (SDSA), break-induced replication (BIR), and error-free lesion bypass pathways. Each of these pathways involves the coupling of a HR event to DNA synthesis. We highlight two major classes of accessory proteins in recombination and replication that facilitate HDR: Recombination mediator proteins exemplified by T4 UvsY, Saccharomyces cerevisiae Rad52, and human BRCA2; and DNA helicases/translocases exemplified by T4 Gp41/Gp59, E. coli DnaB and PriA, and eukaryotic Mcm2-7, Rad54, and Mph1. We illustrate how these factors help to direct the flow of DNA and protein-DNA intermediates on the pathway from a double-strand break or stalled replication fork to a high-fidelity recombination-dependent replication apparatus that can accurately repair the damage. PMID- 21647942 TI - Phytoestrogen calycosin-7-O-beta-D-glucopyranoside ameliorates advanced glycation end products-induced HUVEC damage. AB - Vasculopathy including endothelial cell (EC) apoptosis and inflammation contributes to the high incidence of stroke and myocardial infarction in diabetic patients. The aim of the present study was to investigate the effect of calycosin 7-O-beta-D-glucopyranoside (CG), a phytoestrogen, on advanced glycation end products (AGEs)-induced HUVEC damage. We observed that CG can significantly ameliorate AGEs-induced HUVEC oxidative stress and apoptosis. The ratio of SOD/MDA was significantly increased to the normal level by CG pretreatment. CG preincubation dramatically increased anti-apoptotic Bcl-2 while decreased pro apoptotic Bax and Bad expressions as detected by immunocytochemistry. Moreover, CG ameliorated macrophage migration and adhesion to HUVEC; the monocyte chemotactic protein-1 and interleukin-6 levels in the culture supernatant were dramatically reduced by CG as determined by ELISA; the expressions of inflammatory proteins including ICAM-1, TGF-beta1, and RAGE in both protein and mRNA levels were significantly reduced to the normal level by CG pretreatment as determined by immunocytochemistry and real-time RT-PCR. The intracellular investigation suggests that CG can reverse AGEs-activated ERK1/2 and NF-kappaB phosphorylation, in which estrogen receptors were involved in. Our results strongly indicate that CG can modulate EC dysfunction by ameliorating AGEs induced cell apoptosis and inflammation. PMID- 21647943 TI - Single-substance and mixture toxicity of five pharmaceuticals and personal care products to marine periphyton communities. AB - The single-substance and mixture toxicity of five pharmaceuticals and personal care products (fluoxetine, propranolol, triclosan, zinc-pyrithione, and clotrimazole) to marine microalgal communities (periphyton) was investigated. All compounds proved to be toxic, with median effective concentration values (EC50s) between 1,800 nmol/L (triclosan) and 7.2 nmol/L (Zn-pyrithione). With an EC50 of 356 nmol/L, the toxicity of the mixture falls into this span, indicating the absence of strong synergisms or antagonisms. In fact, a comparison with mixture toxicity predictions by the classical mixture concepts of concentration addition and independent action showed a good predictability in the upper effect range. However, the mixture provoked stimulating effects (hormesis) in the lower effect range, hampering the application of either concept. An independent repetition of the mixture experiment resulted in a principally similar concentration-response curve, again with clear hormesis effects in the lower range of test concentrations. However, the curve was shifted toward higher effect concentrations (EC50 1,070 nmol/L), which likely is due to changes in the initial species composition. Clear mixture effects were observed even when all five components were present only at their individual no-observed-effect concentrations (NOECs). These results show that, even with respect to mixtures of chemically and functionally dissimilar compounds, such as the five pharmaceuticals and personal care products investigated, environmental quality standards must take possible mixture effects from low-effect concentrations of individual compounds into consideration. PMID- 21647944 TI - Dechlorane Plus in house dust from E-waste recycling and urban areas in South China: sources, degradation, and human exposure. AB - Dechlorane Plus (DP) was measured in house dust from e-waste recycling and from urban and rural areas of South China, with geometric mean concentrations of 604, 14.5, and 2.89 ng/g, respectively. Dechlorane Plus in house dust in the e-waste area originated from e-waste recycling activities, whereas household appliances served as a major source of DP in urban house dust. The isomer ratios (f(anti) ) of DP in most dust samples from the e-waste area were significantly lower than those in the urban and rural dust samples and the commercial mixture. Several [ 1Cl + H] and [-2Cl + 2H] dechloro-DPs were identified in house dust from the e waste area, and an a-Cl(11) DP was qualified with concentrations of <55.1 ng/g. Photolytic degradation experiments were conducted by exposing anti-DP, syn-DP, and commercial DP solutions to ultraviolet (UV) light. The slight difference in isomeric half-life derived by photodegradation, as well as the lower f(anti) values in the e-waste combusted residue, suggest a significant influence of isomer-specific thermal degradation of DP during e-waste burning on isomer composition in house dust in the e-waste area. The average estimated daily intakes (EDIs) of DP via house dust ranged from 0.06 to 30.2 ng/d for adults and 0.14 to 121 ng/d for toddlers in the studied area. The average EDIs of a-Cl(11) DP for adults and toddlers in the e-waste area were 0.07 and 0.18 ng/d, respectively. PMID- 21647945 TI - Evidence for behavioral preference toward environmental concentrations of urban use herbicides in a model adult fish. AB - Fish live in waters of contaminant flux. In three urban, fish-bearing waterways of British Columbia, Canada, we found the active ingredients of WeedEx, KillEx, and Roundup herbicide formulations (2,4-D, dicamba, glyphosate, and mecoprop) at low to high ng/L concentrations (0.26 to 309 ng/L) in routine conditions, i.e., no rain for at least one week. Following rain, these concentrations increased by an average of eightfold, suggesting runoff as a major route of herbicide introduction in these waterways. To determine whether fish might be able to limit point-source exposures through sensory-driven behaviors, we introduced pulses of representative herbicide mixtures to individual adult zebrafish (a model species) in flow-through tanks. Fish did the opposite of limit exposure; they chose to spend more time in pulses of herbicide mixtures representative of those that may occur with rain events. This attraction response was not altered by a previous 4 d exposure to lower concentrations of the mixtures, suggesting fish will not learn from previous exposures. However, previous exposures did alter an attraction response to an amino acid prevalent in food (L-alanine). The present study demonstrates that fish living within urban waterways may elect to place themselves in herbicide-contaminated environments and that these exposures may alter their behavioral responses to cues necessary for survival. PMID- 21647946 TI - Landscape-level patterns of mercury contamination of fish in North Texas, USA. AB - Mercury (Hg) is a toxic metal that is found in aquatic food webs and is hazardous to humans. An emerging conceptual model predicts that the areas of the landscape that have the potential to contain food webs with elevated concentrations of Hg are those that receive high amounts of Hg and sulfate deposition and have high coverage of forests and wetlands and low coverage of agriculture. The objective of the present study was to test this conceptual model using concentrations of Hg in largemouth bass (Micropterus salmoides) from 145 reservoirs in four ecoregions of North Texas. The highest level of Hg contamination in fish was in the South Central Plains, the ecoregion that receives the highest levels of Hg and sulfate deposition and contains extensive forest and wetland habitat and little agriculture. The present study has important implications for other areas of the United States, because the South Central Plains extend into parts of Oklahoma, Louisiana, and Arkansas, covering a total area of 152,132 km(2) of the southern United States. PMID- 21647947 TI - Effects of the antihistamine diphenhydramine on selected aquatic organisms. AB - In recent years pharmaceuticals have been detected in aquatic systems receiving discharges of municipal and industrial effluents. Although diphenhydramine (DPH) has been reported in water, sediment, and fish tissue, an understanding of its impacts on aquatic organisms is lacking. Diphenhydramine has multiple modes of action (MOA) targeting the histamine H1, acetylcholine (ACh), and 5-HT reuptake transporter receptors, and as such is used in hundreds of pharmaceutical formulations. The primary objective of this study was to develop a baseline aquatic toxicological understanding of DPH using standard acute and subchronic methodologies with common aquatic plant, invertebrate, and fish models. A secondary objective was to test the utility of leveraging mammalian pharmacology information to predict aquatic toxicity thresholds. The plant model, Lemna gibba, was not adversely affected at exposures as high as 10 mg/L. In the fish model, Pimephales promelas, pH affected acute toxicity thresholds and feeding behavior was more sensitive (no-observed-effect concentration = 2.8 ug/L) than standardized survival or growth endpoints. This response threshold was slightly underpredicted using a novel plasma partitioning approach and a mammalian pharmacological potency model. Interestingly, results from both acute mortality and subchronic reproduction studies indicated that the model aquatic invertebrate, Daphnia magna, was more sensitive to DPH than the fish model. These responses suggest that DPH may exert toxicity in Daphnia through ACh and histamine MOAs. The D. magna reproduction no-observed-effect concentration of 0.8 ug/L is environmentally relevant and suggests that additional studies of more potent antihistamines and antihistamine mixtures are warranted. PMID- 21647948 TI - Using ptilochronology to determine daily mercury deposition in feathers of nestling waterbirds. AB - Feathers are commonly used biomarkers of mercury (Hg) contamination in waterbird species. Most studies that analyze waterbird feathers for Hg content report concentrations on a per-unit mass basis. While this is appropriate for intraspecific comparisons, we suggest a more effective method for studies comparing multiple species of similar size and with similar foraging habits. Ptilochronology is a technique for determining the rate of feather growth in individuals based on their nutritional condition. When paired with Hg analysis, feather growth rates can be used to calculate the average daily Hg deposition into a feather. In this study we used this technique in comparison with the commonly used metric of Hg per-unit feather mass in two waterbird species. Average daily Hg deposition into feathers was 26.7% more sensitive to differences in Hg between the two species, suggesting that this may be a more biologically meaningful metric to use in interspecific comparisons. PMID- 21647949 TI - PCB and organochlorine pesticides in northern fulmars (Fulmarus glacialis) from a High Arctic colony: chemical exposure, fate, and transfer to predators. AB - Organochlorine contaminant concentrations, associated fugacities, and stable isotopes of nitrogen (delta(15) N) are reported for liver, whole body homogenate, and opportunistically collected samples of prey (amphipods), stomach oils, digestive tract contents, and guano for northern fulmars (Fulmarus glacialis) collected at Cape Vera, Devon Island in the Canadian High Arctic. Liver concentrations of polychlorinated biphenyls (SigmaPCB) and SigmaDDT were on average 49.9 +/- 35.4 ng g(-1) and 29.9 +/- 25.2 ng g(-1) wet weight, respectively. Whole body homogenate concentrations of SigmaPCB and SigmaDDT were 637 +/- 293 ng g(-1) and 365 +/- 212 ng g(-1) wet weight, respectively. A mass and energy balance showed that whole body contaminant concentrations, which are seldom reported for Arctic seabirds, are critical in determining contaminant exposure and associated risk to predators such as the Arctic fox (Alopex lagopus). Biomagnification in the fulmars is evident, because concentrations and fugacities of contaminants were generally one to three orders of magnitude higher than those of likely prey items. The fate of diet-derived contaminants along the digestive tract is discussed, in particular with respect to stomach oils, which are used to feed chicks and for defensive purposes. The benefits of considering both concentrations and fugacities are demonstrated and provide information on the absorption and distribution of chemicals within the fulmars and contaminant transfer to offspring and predators. PMID- 21647950 TI - Hematopoietic progenitor cell count, but not immature platelet fraction value, predicts successful harvest of autologous peripheral blood stem cells. AB - Mobilized stem cells in the peripheral blood (PB) must be efficiently harvested at the appropriate time before autologous PB stem cell (PBSC) transplantation. Enumeration of CD34+ cells in the PB before apheresis predicts the number of PBSCs that can be collected, but the cytometric techniques used are complex and expensive. Therefore, it is necessary to identify an alternative to the CD34+ cell count in PBSC harvest-time monitoring. Fully automated flow cytometry using blood cell counters now allows reliable quantification of immature myeloid cells in the PB, referred to as hematopoietic progenitor cells (HPC), and reticulated platelets, expressed as the immature platelet fraction (IPF). Immature or reticulated platelets are thought to correlate with thrombopoietic activity of the marrow. Following a chemotherapy nadir, the recovery of white blood cell and platelet counts has been used to determine the right time for apheresis. Therefore, we examined whether the HPC count and IPF value could be used to predict PBSC mobilization in 20 patients with hematological malignancies. The HPC count was found to be correlated with the CD34+ cell count (r = 0.84, P < 0.01), whereas the IPF value was not (r = 0.37, P = 0.44). Therefore, the HPC count, but not the IPF value, is a possible predictor of the timing of autologous stem cell transplantation. PMID- 21647951 TI - Predicting hematopoietic stem cell mobilization failure in patients with multiple myeloma: a simple method using day 1 CD34+ cell yield. AB - Early and reliable prediction of the likelihood of achieving adequate stem cell collection for autologous stem cell transplantation (ASCT) in patients with multiple myeloma (MM) would improve collection efficiency, prevent unnecessary aphereses, and permit appropriate treatment alterations. No previous study has reported a threshold CD34+ cell collection quantity on Day 1 or 2 of leukapheresis that could predict successful stem cell collection. We performed a retrospective analysis of all MM patients undergoing first attempt of stem cell collection at our institution from 2001 through 2008. Recursive partitioning analysis was used to identify Day 1 or Day 1+2 CD34+ collection quantity that predicted failure to reach target >= 2 * 10(6) CD34+ cells/kg within five days of collection. Totally, 172 patients were included in the analysis. Patients underwent mobilization with G-CSF or G-CSF+ chemotherapy. 23 of 172 patients (13.4%) failed to collect sufficient (>= 2 * 10(6) CD34+ cells/kg) CD34+ cells after five days of apheresis: 22 of 29 who collected <= 0.70 * 10(6) CD34+ cells/kg and 1 of 143 who collected > 0.70 * 10(6) CD34+ cells/kg (75.9% vs. 0.7%, P < 0.001) on Day 1. Collection failure occurred in 23 of 30 patients who collected <= 1.54 * 10(6) CD34+ cells/kg and 0 of 142 who collected >1.54 * 10(6) CD34+ cells/kg (76.7% vs. 0%, P < 0.001) on Days 1 + 2. Day 1 CD34+ cell collection quantity identifies patients unlikely to achieve adequate collection for ASCT. Patients who collect <= 0.70 * 10(6) CD34+ cells/kg on day 1 could be considered for treatment modifications to improve CD34+ collection, such as early administration of plerixafor or large volume apheresis. PMID- 21647952 TI - Update on extracorporeal photopheresis in heart and lung transplantation. AB - Transplant rejection of solid organs remains a threat to thousands of patients despite modern immunosuppressive regimens. The currently available drugs are associated with severe complications such as hypertension, diabetes mellitus, renal failure, risk of infections, and malignancies among many others and, often enough, still allow episodes of rejection. New and less-toxic immunologic measures are desperately needed to accomplish the desired tolerance to the transplant without the undesirable side effects. Extracorporeal photopheresis (ECP) has been shown to benefit especially patients with cardiac transplants, but also those who received lung allografts. ECP likely modulates the recipient's antigen-specific immune responses and inflammation in transplantation by in vivo generation of apoptotic leukocytes. This review will highlight the need for ECP, how it is thought to act, and the published evidence for its role in cardiac and pulmonary transplantation. PMID- 21647953 TI - Stroke in a young patient treated by alteplase heralding an acquired thrombotic thrombocytopenic purpura. AB - BACKGROUND AND PURPOSE: Thrombotic thrombocytopenic purpura (TTP) is a life threatening multisystem disorder characterized by thrombocytopenia and fluctuating neurological symptoms due to microinfarcts. In rare cases, large cerebral arteries can be occluded. SUMMARY OF THE CASE: We report on a 30-year old woman with a first-ever acute stroke related to a right proximal MCA M1 occlusion. Platelet count was normal at admission and progressively decreased 6 days after intravenous thrombolysis with the occurrence of a hemolytic anemia with schistocytes. Most biological anomalies reversed after plasma exchange. No hemorrhagic complication occurred. Diagnosis of initial TTP was confirmed by low ADAMTS13 activity and positivity of anti-ADAMTS13 antibody. CONCLUSION: This observation highlights the fact that even if platelet count and hemoglobin rate are normal in the beginning, an acute ischemic stroke in a young patient can be related to TTP. Faced with subsequent thrombopenia, practitioners should be aware of acquired TTP, and, thus, schistocytes, haptoglobin, and LDH assays should be performed. Early diagnosis is paramount to start the life-saving plasma exchanges. PMID- 21647954 TI - Management of cyclosporine overdose in a hematopoietic stem cell transplant patient with sequential plasma exchange and red blood cell exchange. AB - Cyclosporine is commonly used as an immunosuppressive agent in both solid organ and bone marrow transplant. While used for graft rejection in organ transplantation, cyclosporine has been used to enable tolerance and for prevention of acute graft-versus-host disease in bone marrow transplant [Ratanatharathorn et al., Blood 1998;92:2303-2314]. Cyclosporine has a narrow therapeutic window, and many patients develop some level of toxicity even within the therapeutic range. Common toxicities include hypertension, nephrotoxicity, electrolyte abnormalities, hyperglycemia, and neurotoxicity [Woo et al., Bone Marrow Transplant 1997;20:1095-1098]. Management of cyclosporine toxicity is not clearly defined and is primarily supportive in nature. In cases of significant elevations of cyclosporine levels, limited data are available but suggest that whole blood exchange may be effective [Kwon et al., J Heart Lung Transplant 2006;25:483-485; Leitner et al., Transplantation 2003;75:1764-1765]. We present a case of successful rapid clearance of cyclosporine utilizing a combined approach of red cell exchange and plasma exchange. PMID- 21647955 TI - Comparative analysis of serum proteomes of degenerative scoliosis. AB - Degenerative scoliosis (DS) is an important degenerative lumbar disease causing spinal dysfunction and affecting the quality of life of the elderly, and is associated not only with severe back or leg pain but also with complicated surgical outcomes. The pathogenesis of DS is still unknown. Therefore, it is very important to ascertain the etiology of degenerative scoliosis and establish related molecular markers predicting and controlling the scoliosis. For the first time, we used two-dimensional fluorescence DIGE to compare the serum proteome profiles of 12 DS patients and controls. Proteins found to be differentially expressed were identified by MALDI-TOF mass spectrometric analysis, coupled with database interrogation. Eleven spots that were differentially expressed in the sera of DS patients were found, and eight gene products were identified among these spots. Clusterin, CLU cDNA FLJ57622, ALB cDNA FLJ50830, Hypothetical short protein, HLA-A MHC class 1 antigen. (Fragment), ALB 23 kDa protein, Isoform 1 of G protein-regulated inducer of neurite outgrowth 1 (GPRIN I)and Ficolin-3 were down-regulated in the sera of DS patients. The decreased levels of Clusterin and Ficolin-3 were confirmed by Western blot. The information obtained with this proteomic analysis will be very useful in understanding the pathophysiology of DS as well as in finding candidates as drug targets of DS. These results may provide a novel approach for the pathogenesis study of DS. PMID- 21647956 TI - Lubricin distribution in the torn human anterior cruciate ligament and meniscus. AB - The objective of this study was to: (1) determine the distribution of lubricin in the human torn anterior cruciate ligament (ACL) and meniscus; (2) determine the distribution of lubricin in the human intact ACL and meniscus; (3) and identify potential cellular sources of lubricin in these tissues. Ten torn ACLs and six torn menisci were obtained from surgeries; for comparison, 11 intact ACLs and 13 intact menisci were obtained from total knee replacements. Samples were formalin fixed and processed for immunohistochemical staining with a monoclonal antibody for lubricin. In torn ACLs and menisci, lubricin was generally found as a discrete layer covering the torn surface. No surface lubricin staining was found on the transected edges produced during excision. Lubricin was also found on the native surfaces of intact ACLs and menisci. In all tissues, lubricin was found in the matrix and intracellularly. The surface layer of lubricin coating torn edges of ACLs and menisci may interfere with the integrative healing process needed for repair. PMID- 21647957 TI - Biomechanical examination of the thoracic spine--the axial rotation moment and vertical loading capacity of the transverse process. AB - Our objective was to examine the load-bearing capacity of the transverse processes of human cadaveric thoracic vertebrae to vertical loads and axial rotation moments (i.e., moment applied in the transverse plane). A secondary objective was to examine the effect of the attached rib stumps. We wanted to demonstrate that the transverse process is durable enough to support the CAB hook -a complementary hook to the CD system--and can handle the vertical load or axial rotation moment during correction of scoliosis. We used 107 thoracic vertebrae removed from 10 cadavers. They were prepared in vertebral pairs, and were fixed into a material testing apparatus. Superoinferior vertical loads and axial rotation moments were applied to the transverse process using the CAB hooks at a rate of 30 mm/min and 8.5 degrees /s respectively until it fractured. We recorded 142 measurements, 99 were for vertical load and 43 for axial rotation moment. The average ultimate vertical load was 338 (SD = 128) N and the average ultimate axial rotation moment was 14.4 (SD = 4.52) Nm. The ultimate axial rotation moment for specimens with rib stumps attached was significantly greater than for specimens without rib stumps 15.9 (SD = 4.1) Nm versus 12.5 (SD = 4.4) Nm. Our results showed that both the vertical and axial rotation loading capability of the transverse process are large enough to withstand significant correctional forces, without fracture, through the CAB hooks. PMID- 21647958 TI - Microstructure of the vocal fold in elderly humans. AB - Significant changes in the voice occur after the age of 50 years. Changes in the structure of the vocal fold (VF) can interfere with the voice. The aim of this study is to investigate the structure of the VF of elderly people that may contribute to the tendency of the human voice to deteriorate. Larynges were obtained from eight embalmed cadavers aged 72-98 years. The middle portion of each vocal fold was removed and placed in 4% buffered formalin. Tissue blocks were then processed and embedded in wax. Four to six micron coronal sections were cut and stained with Haematoxylin and Eosin (H&E), Masson's Trichrome (MTS), Elastin van Gieson (EVG), Herovici (HERO), picrosirius and CD31 immunohistochemical marker in order to study the collagen fibers, elastic fibers and microvasculature of the VF. The maturity of collagen fibers within the VF were noted to increase from the superficial to the deep layer of the lamina propria (LP). Contrary to current literature, the amount of elastic fibers was sparse in the superficial layer of the LP in the vocal tissue of elderly cadavers. Numerous cross-sectioned blood vessels were seen in the lamina propria near the free edge, and near the superior and inferior surfaces of the VF. The presence of lymphatic vessels was confirmed in the VF of elderly subjects. This study revealed that the collagenous component of the deep layer of the VF LP was made up of mature fibers whilst immature collagen fibers made up the superficial layer of the LP. There was a notable scarcity of elastic fibers in the superficial layer of the LP. Lymphatics were seen and were orientated differently in the geriatric vocal folds. PMID- 21647959 TI - Supernumerary and absent limbs and digits of the lower limb: a review of the literature. AB - Anatomical history over centuries includes description of a wide variety of malformations involving the lower limbs. This article offers an organized review of these diverse abnormalities, including new understanding of mechanisms through recent discoveries in genetics and molecular biology. In 19th century Europe, a number of unique anomalies were reported, as well as evidence of foot amputations occurring in ancient Peruvian culture. Embryologically, the limbs develop early, with the lower limb being recognizable for the first time at stage 13 of development. By stage 23, the toes are clearly defined and by birth, although the legs appear bowed, the tibia and fibula are straight. Removal of the apical ectodermal ridge results in cessation of limb development, conversely, a second apical ectodermal ridge results in duplication of distal structures. Supernumerary limbs have been documented to occur as part of a teratoma with unique morphology and accompanying blood supply. Additionally, many examples of polydactyly occur in the foot postulating that deletion of chromosome 22q11 is involved in postaxial polydactyly. Such deletions occur near the middle of the chromosome at a location designated q11.2 (i.e., on the long arm of one of the pair of chromosomes 22) and this syndrome is also referred to as DiGeorge syndrome, which has a prevalence estimated at 1:4,000. Absence of the lower limbs has also been noted, with hypoplasia of the fibula being the most common manifestation of congenital bone absences in the lower limb. In addition to fibular aplasia, cases of tibial aplasia have been reported. This article is important for surgeons attempting correctional repair of lower limb anomalies, as well as providing analysis of the historical, anatomical and clinical aspects of supernumerary and absent limbs and digits for the lower limb. PMID- 21647960 TI - Influence of food consistency on growth and morphology of the mandibular condyle. AB - The objective of this study was to determine if variation in the shape and mineralization of the mandibular condyle are the result of natural adaptation in response to different functional loading demands. Eight female Kuni Kuni piglets were randomly assigned to two groups of four, receiving either a soft or hard diet. Each animal was given three separate doses of vital stains intravenously at set time points during the study. At 8.5 months, animals were euthanized and temporomandibular joints (TMJs) were excised. Histological analysis was used to measure the amount of new bone deposition in the anterior, central, and posterior regions of the mandibular condyle. Backscatter electron (BSE) imaging was used as a semiquantitative estimate of bone mineralization in these two diet groups. Histology revealed that the degree of new bone deposition in the hard-diet group was significantly (n = 4, P < 0.001, paired t-test) higher than that of the soft diet group. Also, the majority (87%) of animals fed a hard diet tended to show greater new bone deposition on the leftside in comparison to the right, indicating a chewing preference for the left side. In both groups, the degree of new bone deposition was significantly (P < 0.01) higher in the posterior area than in other regions. BSE imaging corroborated basic histology results, with significantly (P < 0.01) higher mineralization levels detected in the hard-diet group. These findings indicate that diet consistency has a small but significant effect on the rate of bone deposition in the mandibular condyle. PMID- 21647961 TI - Fetal anatomy of the lower cervical and upper thoracic fasciae with special reference to the prevertebral fascial structures including the suprapleural membrane. AB - The aim of this study was to find basic rules governing the fetal anatomy of the deep cervical fasciae and their connections to the mediastinal fasciae. We examined the histology of paraffin-embedded preparations of 18 mid-term fetuses (5 between 9 and 12 weeks of gestation, 3 between 15 and 18 weeks, and 10 between 20 and 25 weeks). The prevertebral lamina of the deep cervical fasciae (PLDCF) developed as an intermediate aponeurosis for the bilateral bellies of the longus colli muscles. In contrast, the alar fascia developed as a connecting band between the bilateral adventitiae of the common carotid artery. The retropharyngeal fascia became evident much later than the latter two fasciae. The fascia covering the thymus was thicker than the fascia for the strap muscles (the pretracheal lamina of the cervical fascia). The primitive suprapleural membrane, or Sibson's fascia, contained veins and fatty tissues, and was composed of the alar fascia rather than the PLDCF, tranversalis fascia, or endothoracic fascia. The prevertebral two-laminar configuration was rather evident in the early stages of development because, in the later stages, the fasciae together provided a multilaminar structure, especially in the lateral area in front of the longus colli, which suspended the cupula pleurae. To consider a continuation from the base of the neck to the upper mediastinum, the alar fascia seems to be a key structure for connecting the vascular sheath to the parietal pleura. PMID- 21647962 TI - A comprehensive analysis of muscle recruitment patterns during shoulder flexion: an electromyographic study. AB - Although flexion is a common component of the routine clinical assessment of the shoulder the muscle recruitment patterns during this movement are not clearly understood making valid interpretation of potential muscle dysfunction problematic. The purpose of this study was to comprehensively examine shoulder muscle activity during flexion in order to compare the activity levels and recruitment patterns of shoulder flexor, scapular lateral rotator and rotator cuff muscles. Electromyographic (EMG) data were recorded from 12 shoulder muscles sites in 15 volunteers. Flexion was performed in standing in the sagittal plane at no load, 20%, and 60% of each subject's maximum load. EMG data were normalized to maximum values obtained during maximum voluntary contractions. Results indicated that anterior deltoid, pectoralis major, supraspinatus, infraspinatus, serratus anterior, upper, and lower trapezius were activated at similar moderate levels. However, subscapularis was activated at low levels and significantly lower than supraspinatus and infraspinatus. Similar activity patterns across time were demonstrated in the muscles that produce flexion torque, laterally rotate the scapula, as well as supraspinatus and infraspinatus, and did not change as flexion load increased. The onset of activity in supraspinatus and anterior deltoid occurred at the same time and prior to movement of the limb at all loads with infraspinatus activity also occurring prior to movement onset at the medium and high load conditions only. Posterior rotator cuff muscles appear to be counterbalancing anterior translational forces produced during flexion and it would appear that supraspinatus is one of the muscles that consistently "initiates" flexion. PMID- 21647963 TI - Persistent median artery: cadaveric study and review of the literature. AB - The persistent median artery (PMA) may compress the median nerve (MN) and may be a significant supply of blood to the hand. Two cases of unilateral PMA (4%) were detected during the dissection of 50 upper limbs. The first case was a 75-year old, right-handed male who suffered from chronic pain in both upper limbs, especially the left side. A dissection of his left upper limb revealed a PMA piercing both the MN and the medial branch of the anterior interosseous nerve. This artery coursed distally, deep to the transverse carpal ligament (TCL), forming a median-ulnar pattern for the superficial palmar arch (SPA). The PMA was superficial to two nerves at the distal edge of the TCL; the extraligamentous recurrent thenar (RT) branch of the MN and the third common digital nerve (TCDN). The second case was from the left side of an 80-year-old female found to have a high origin of the radial artery with trifurcation of the latter into PMA, common interosseous, and ulnar arteries. The PMA passed deep to the TCL forming a radial median-ulnar pattern of SPA. Both the transligamentous RT branch of the MN and the TCDN passed deep to the PMA inside the carpal tunnel, before the abnormal crossing of the latter nerve ventral to the SPA on its way to the digits. The relationships of the PMA to various MN branches may have important implications regarding the diagnosis and treatment of MN compressive neuropathies. PMID- 21647964 TI - Use of MRI images to measure tissue thickness over the ischial tuberosity at different hip flexion. AB - The goal of this experiment was to investigate changes in the thickness of the soft tissue overlying the ischial tuberosity (IT) due to changes in hip flexion angle and the addition of a sitting load. Eleven healthy subjects were tested. An apparatus constructed from foam blocks and an air bladder was used to position the subjects in different postures within an MRI tube. MRI images of the buttocks and thigh were obtained for four postures: Supine, 45 degrees Hip Flexion, Non Weight-Bearing 90 degrees Hip-Flexion, and Weight-Bearing 90 degrees Hip Flexion. The thickness of muscle, adipose tissue, and skin was measured between the IT tip and skin surface, perpendicular to the cushion placed beneath the thighs. The tissue overlying the IT was found to be significantly (P < 0.001) thinner in 90 degrees Hip-Flexion (73.8 +/- 9.0 mm) than in the supine position (135.9 +/- 8.1 mm). Muscle thickness decreased significantly from Supine to Non Weight-Bearing 90 degrees Hip-Flexion (59.1 +/- 8.5%, P < 0.001), and further decreased from Non-Weight-Bearing to Weight-Bearing 90 degrees Hip-Flexion (46.2 +/- 7.9%, P < 0.001). Under Weight-Bearing 90 degrees Hip-Flexion, the muscle tissue deformed significantly (P < 0.001) more than the adipose tissue and skin. We concluded that the tissue thickness covering the IT significantly decreased with hip flexion, and further decreased by nearly half during loading caused by sitting. In addition, the muscle tissue experienced the largest deformation during sitting. The results of this study may improve our understanding of risk factors for pressure ulcer development due to changes in tissue padding over the IT in different postures. PMID- 21647965 TI - Nerve terminal distribution in the human tongue intrinsic muscles: An immunohistochemical study using midterm fetuses. AB - Intrinsic tongue muscles, especially the transverse and vertical (T&V) muscles, regulate the shape of the tongue. However, little information is available on the nerve distribution pattern in human T&V muscles. Using S100 protein immunohistochemistry for paraffin-embedded histology, we investigated semiserial sagittal or frontal sections of eight human fetal tongues (180-240 mm crown-rump length: CRL). The height of the T&V muscle bundle showed a threefold difference between specimens with a small and a large CRL. Thus, the T&V muscles were still growing at the stages examined. In the intrinsic longitudinal muscles and all extrinsic tongue muscles, we observed the typical motor endplate band. In lower magnification views, the T&V muscles also appeared to carry the band in the lateral part of the tongue, where the genioglossus muscle fibers did not cross these muscles. However, in higher magnification views, the nerve terminal distribution in the T&V muscles showed a unique rule: the nerve terminal for the transverse muscle bundle was located distantly from that of the adjacent vertical muscle bundle. This pattern seemed to be established during the stages examined. To provide such "distantly separated nerve terminals," thin nerve twigs took a highly curved course oblique to the T&V muscle bundles. We hypothesize that the unique nerve course and terminal distribution in the T&V muscles are a result of sorting to provide a good functional match between the nerve fiber and the muscle bundle. After sorting, the T&V muscle cells may initiate proliferation to increase the muscle bundle. PMID- 21647966 TI - A proposed explanation for the development of the torus palatinus. PMID- 21647967 TI - Lumbar shape characterization of the neural arch and vertebral body in spondylolysis: A comparative skeletal study. AB - The lumbar vertebral shape is an important causative factor in spondylolysis (SP). However, a complete characterization of this shape, that would shed light on its pathophysiology enhance its earlier diagnosis, is still missing. The aim of this study was to evaluate the shape distinctiveness of the lumbar (L1-L5) vertebral body (VB) and neural arch (isthmus, lamina, vertebral canal, spinous, and transverse processes) in SP. Using a three-dimensional (3D) digitizer, the VB length, width, height, and sagittal wedging as well as the lengths of the isthmus, lamina spinous, and transverse processes of all lumbar vertebrae (L1-L5) were measured from 115 male skeletons with bilateral SP at L5 and compared with 120 normal ones. Compared with the normal group, the following results were indicated in SP [analysis of variance (ANOVA), P < 0.05]: the VB's are significantly longer at the superior surfaces of L1-L4 and inferior surfaces of L1-L3 (total mean difference = 7.61 mm and 3.94 mm, respectively), and wider at the superior surfaces of L1-L4 and inferior surfaces of L1-L5 (total mean difference = 10.06 mm and 12.90 mm, respectively); The L5 VB is significantly shorter posteriorly (mean difference = 3.05 mm) and more lordotic (+1 degrees of dorsal wedging); L5 is manifested with longer lamina (mean difference = 1.85-2.18 mm), longer isthmus (mean difference = 3.24-4.69 mm), and longer and wider vertebral canal (mean differences = 3.64 mm and 1.13 mm, respectively); The L2-L5 spinous processes are significantly longer (total mean difference = 7.45 mm). Clinically, detecting the aforementioned unique lumbar vertebral morphometric features in individuals suffering from chronic low back pain may facilitate the early diagnosis of SP. PMID- 21647968 TI - Topographical anatomy of central venous system in extremely low-birth weight neonates less than 1000 grams and the effect of central venous catheter placement. AB - Central venous catheterization is widely used in neonatology. Although ultrasonic guidance for central venous catheter placement is available, complications occur significantly more frequently in infants, especially neonates, than in adults. This study seeks to determine the characteristics, topographical conditions, regional relationships, and diameters of the venous structures of the upper extremity and the thoracic central venous system in extremely small preterm neonates (mean: 900 g). Nine formaldehyde-fixed preterm stillborns were prepared (mean 27 2/7 weeks' gestational age). The anatomical preparation involved the complete thoracic wall, neck and shoulder region, and preparation of the upper extremities. It was shown that the course of the internal jugular vein can be influenced by rotation of the head. Maximum head rotation (80 degrees ) to the contralateral side leads the internal jugular vein to overlap the common carotid artery and sharpens the confluence angle of the internal jugular into the brachiocephalic vein. We propose that this has the potential to result in dislocation of the catheter. Less rotation of the head (<30 degrees ) is favorable as the internal jugular vein and common carotid artery run in parallel. Commonly used central venous catheters (2F-4F) may not occlude the vascular lumen completely. Small central venous cannulation using a single-orifice catheter through arm veins (1F) may also not occlude peripheral vessels of the upper extremity (cephalic and basilic veins). The right internal jugular vein has a straight course, appears suitable for central venous access and less hazardous, especially when using stiff catheters. The use of small straight wire guides is recommended. PMID- 21647969 TI - Topography of the arteries supplying the masseter muscle: Using dissection and Sihler's method. AB - Various surgical procedures require surgeons to have detailed knowledge of the course of blood vessels in the masseter muscle, such as masseter muscle flap formation, mandibular angle resection, parotidectomy, and mandibular ramus osteotomy. Without this knowledge serious complications can occur, endangering the lives of patients. Occasionally, during routine dissections we sometimes encounter an additional branch. The purpose of this study was to provide a comprehensive detailed anatomic description of the blood supply of the masseter muscle. This will provide critical information for various surgical procedures. Twenty-five Korean cadavers were dissected and subjected to modified Sihler's method to reveal the branching patterns of the arteries surrounding the masseter muscle, and its intramuscular blood supply. The masseter can be supplied by seven branches from the external carotid artery. Among these, the masseteric branch from the deep temporal artery has not been described previously. This previously undescribed branch enters the medial surface of the masseter, turning medially around the anterior border of the ramus immediately after the branching point of the deep temporal artery. The branch originating from the transverse facial artery was the largest, and the branches originating from the external carotid artery and deep temporal artery were the smallest. Examination of intramuscular patterns revealed that the branches of the transverse facial artery were the most widely distributed. When considering arterial diameters and distribution areas, the branches of the transverse facial artery can be considered the main components of the seven branches supplying the masseter muscle. PMID- 21647971 TI - Materials forum. PMID- 21647970 TI - Post-transcriptional up-regulation of miR-21 by type I collagen. AB - Composition of extracellular matrix (ECM) is crucial to the establishment and maintenance of epithelial apical-basolateral polarity. Increased ECM rigidity caused by deposition of fibrillar collagen, for example, collagen type I (Col-1), promotes loss of epithelial polarity and tumor progression. microRNAs are small non-coding RNAs that regulate gene expression and fundamental cellular processes. The current study explored a link between microRNAs and Col-1 using organotypic three-dimensional culture in which epithelial cells are embedded within Matrigel, a mimic of basement membrane matrix (Matrigel 3-D). Matrigel 3-D culture of A549, MCF-7, and mK-ras-LE cells (lung and mammary epithelial cell lines) gave rise to acinus, an in vitro equivalent of apical-basolateral polarity that consists of a polarized monolayer of epithelial cells facing a central lumen. Supplementation of Col-1 disrupted acinus. Moreover, Col-1 up-regulated the expression of miR-21, a well-documented oncogenic microRNA, via a post-transcriptional mechanism. Similar post-transcriptional up-regulation of miR-21 correlated with deposition of Col-1 in a murine model of lung fibrogenesis. In summary, our findings link altered ECM composition/rigidity and the expression of oncogenic microRNAs. The current study also suggests a novel post-transcriptional mechanism for regulation of miR-21 expression at maturation from pre-miR-21 to mature miR-21. PMID- 21647972 TI - What does materials science learn from conjugated oligomers? PMID- 21647973 TI - Polymer electrolytes for lithium-ion batteries. AB - The motivation for lithium battery development and a discussion of ion conducting polymers as separators begin this review, which includes a short history of polymer electrolyte research, a summary of the major parameters that determine lithium ion transport in polymer matrices, and consequences for solid polymer electrolyte development. Two major strategies for the application of ion conducting polymers as separators in lithium batteries are identified: One is the development of highly conductive materials via the crosslinking of mobile chains to form networks, which are then swollen by lithium salt solutions ("gel electrolytes"). The other is the construction of solid polymer electrolytes (SPEs) with supramolecular architectures, which intrinsically give rise to much enhanced mechanical strength. These materials as yet exhibit relatively common conductivity levels but may be applied as very thin films. Molecular composites based on poly(p-phenylene)- (PPP)-reinforced SPEs are a striking example of this direction. Neither strategy has as yet led to a "breakthrough" with respect to technical application, at least not for electrically powered vehicles. Before being used as separators, the gel electrolytes must be strengthened, while the molecularly reinforced solid polymer electrolytes must demonstrate improved conductivity. PMID- 21647974 TI - Solid-State (2) H NMR Determination of Poly(aniline) Conformation Within a MoO(3) Nanocomposite. PMID- 21647975 TI - Chemical contrast on a microphase-separated block copolymer surface observed by scanning force microscopy. PMID- 21647976 TI - Nature of the metallic state in conducting polypyrrole. PMID- 21647977 TI - Fluorescence chemosensor for metal ions using conjugated polymers. PMID- 21647978 TI - Oxidized thin films of c(60) : a new humidity-sensing material. PMID- 21647979 TI - Metal Alkoxide/Hexa-2,4-diyne-1,6-diol Hybrid Polymers: Synthesis and Use as Precursors to Metal Carbides and Nitrides. PMID- 21647980 TI - Self-assembly of an operating electrical circuit based on shape complementarity and the hydrophobic effect. PMID- 21647981 TI - Controlled mineralization and assembly of hydrolysis-based nanoparticles in organic solvents combining polymer micelles and microwave techniques. PMID- 21647982 TI - The Effect of Terminal Hydrogen Bonding on the Structure and Dynamics of Nanoparticle Self-Assembled Monolayers (SAMs): An NMR Dynamics Study. PMID- 21647983 TI - Control of the Photonic Crystal Properties of fcc-Packed Submicrometer SiO(2) Spheres by Sintering. PMID- 21647984 TI - The synthesis of mesoporous copper silicates using organofunctional silicon alkoxides. PMID- 21647985 TI - Controlled drying: the key to better quality porous semiconductors. PMID- 21647986 TI - Soft and wet conducting polymers for artificial muscles. PMID- 21647987 TI - From mesoscopic to nanoscopic surface structures: lithography with colloid monolayers. PMID- 21647988 TI - Mechanistic studies of Pd-catalyzed regioselective aryl C-H bond functionalization with strained alkenes: origin of regioselectivity. AB - Mechanistic studies of a palladium-catalyzed regioselective aryl C-H functionalization of 2-pyrrole phenyl iodide with norbornene are presented. Kinetic and spectroscopic analyses together with crystallographic data provide evidence for intermediates in a proposed stepwise mechanism. On the basis of the mechanistic studies, the origin of the regioselectivity is due to a ligand exchange between I(-) and HO(-) on the norbornyl palladium complex. These mechanistic studies also implicate that either alkoxide or water is responsible for the formation of the palladacycle, but a reversible ring-opening-ring-closing process of the palladacycle with HX can retard the rate of reaction of a key intermediate. The significant aspects of the proposed mechanism are discussed in detail. PMID- 21647989 TI - Ligand effects in the formation of tertiary carbanions from substituted tertiary aromatic amides. AB - Reaction of 2-isopropyl-(N,N-diisopropyl)-benzamide 5 with tBuLi in ether results in ortho deprotonation and the formation of a hemisolvate based on a tetranuclear dimer of (5-Li(o))(2)?Et(2)O. The solid-state structure exhibits a dimer core in which the amide oxygen atoms fail to stabilize the metal ions but are instead available for interaction with two metalated monomers that reside peripheral to the core. Reaction of 5 with tBuLi in the presence of the tridentate Lewis base PMDTA (N,N,N',N'',N''-pentamethyldiethylenetriamine) takes a different course. In spite of the tertiary aliphatic group at the 2-position in 5, X-ray crystallography revealed that a remarkable benzylic (lateral) deprotonation had occurred, giving the tertiary benzyllithium 5-Li(l)?PMDTA. The solid-state structure reveals that amide coordination and solvation by PMDTA combine to distance the Li(+) ion from the deprotonated alpha-C of the 2-iPr group (3.859(4) A), thus giving an essentially flat tertiary carbanion and a highly distorted aromatic system. DFT analysis suggests that the metal ion resides closer to the carbanion center in solution. In line with this, the same (benzylic) deprotonation is noted if the reaction is attempted in the presence of tridentate diglyme, with X-ray crystallography revealing that the metal is now closer to the tertiary carbanion (2.497(4) A). Electrophilic quenches of lithiated 5 have allowed, for the first time, the formation of quaternary benzylic substituents by lateral lithiation. PMID- 21647990 TI - Asymmetric cycloaddition of beta,gamma-unsaturated alpha-ketoesters with electron rich alkenes catalyzed by a chiral Er(OTf)3/N,N'-dioxide complex: highly enantioselective synthesis of 3,4-dihydro-2H-pyrans. AB - The asymmetric inverse-electron-demand hetero-Diels-Alder (HDA) reactions of beta,gamma-unsaturated alpha-ketoesters with electron-rich alkenes were investigated, with an N,N'-dioxide/erbium(III) complex employed as the catalyst. Quantitative conversion of the beta,gamma-unsaturated alpha-ketoesters and excellent enantioselectivities (up to >99% ee) and diastereoselectivities (up to >99:1 d.r.) were observed for a broad range of substrates by using a 0.5-0.05 mol% catalyst loading under mild reaction conditions. The reaction could be scaled up to the gram scale with the same results. In addition, this was the first application of Er(OTf)(3) to the asymmetric inverse-electron-demand HDA reaction and it behaved as an efficient catalyst. Moreover, the synthetic utility of this methodology was demonstrated with the synthesis of key intermediates of some natural products. PMID- 21647991 TI - Highly trifluoromethylated platinum compounds. AB - The homoleptic, square-planar organoplatinum(II) compound [NBu(4)](2) [Pt(CF(3))(4)] (1) undergoes oxidative addition of CF(3) I under mild conditions to give rise to the octahedral organoplatinum(IV) complex [NBu(4)](2) [Pt(CF(3))(5)I] (2). This highly trifluoromethylated species reacts with Ag(+) salts of weakly coordinating anions in Me(2)CO under a wet-air stream to afford the aquo derivative [NBu(4)][Pt(CF(3))(5) (OH(2))] (4) in around 75% yield. When the reaction of 2 with the same Ag(+) salts is carried out in MeCN, the solvento compound [NBu(4) ][Pt(CF(3))(5)(NCMe)] (5) is obtained in around 80% yield. The aquo ligand in 4 as well as the MeCN ligand in 5 are labile and can be cleanly replaced by neutral and anionic ligands to furnish a series of pentakis(trifluoromethyl)platinate(IV) compounds with formulae [NBu(4)][Pt(CF(3))(5) (L)] (L=CO (6), pyridine (py; 7), tetrahydrothiophene (tht; 8)) and [NBu(4)](2) [Pt(CF(3))(5)X] (X=Cl (9), Br (10)). The unusual carbonyl platinum(IV) derivative [NBu(4)][Pt(CF(3))(5) (CO)] (6) is thermally stable and has a nu(CO) of 2194 cm(-1). The crystal structures of 2?CH(2)Cl(2), 5, [PPh(4) ][Pt(CF(3))(5)(CO)] (6'), and 7 have been established by X-ray diffraction methods. Compound 2 has shown itself to be a convenient entry to the chemistry of highly trifluoromethylated platinum compounds. To the best of our knowledge, compounds 2 and 4-10 are the organoelement compounds with the highest CF(3) content to have been isolated and adequately characterized to date. PMID- 21647992 TI - A zinc-sensing glucose-based naphthyl imino conjugate as a detecting agent for inorganic and organic phosphates, including DNA. PMID- 21647993 TI - Columnar benzoperylene-hexa- and tetracarboxylic imides and esters: synthesis, mesophase stabilisation and observation of charge-transfer interactions between electron-donating esters and electron-accepting imides. AB - Benzo[ghi]perylene 1,2,4,5,10,11-hexacarboxylic trialkylimide and dialkylimido dialkyl ester derivatives, displaying a thermodynamically stable hexagonal columnar liquid-crystalline phase at room temperature, have been obtained by the use of previously unexplored chiral racemic alpha-branched alkylimide functions. One of the trialkylimides described here is the first room temperature columnar solely oligo-alkylimide-substituted arene, and thus constitutes a prototype case of self-assembling organic acceptor materials. As the related hexacarboxylic hexaesters are found to exhibit only a weak tendency to form columnar mesophases, benzo[ghi]perylene 1,2,5,10-tetracarboxylic tetraalkyl esters have been synthesized by regioselective oxidative Diels-Alder addition of maleic anhydride to 3,10-dicyanoperylene, and a room temperature hexagonal columnar mesophase was obtained with branched alkyl chains. The acceptor-type electronic properties of the tri- and diimides have been found to be considerably more pronounced than those of the hexa- and tetracarboxylic esters, and to approach those of the prototype acceptor material C(60). The formation of bathochromically absorbing donor-acceptor complexes was observed with a di- or triimide as acceptor and a tetraester as donor, but not with a hexaester as donor. Exploiting the non negligible differences in reduction and oxidation potentials between all four types of materials, the minimum HOMO energy difference necessary for charge transfer-complex formation has been determined to lie between 0.29 and 0.35 eV. PMID- 21647994 TI - Efficient epoxidation of electron-deficient alkenes with hydrogen peroxide catalyzed by [gamma-PW10O38V2(MU-OH)2]3-. AB - A divanadium-substituted phosphotungstate, [gamma-PW(10)O(38)V(2)(MU-OH)(2)](3-) (I), showed the highest catalytic activity for the H(2)O(2)-based epoxidation of allyl acetate among vanadium and tungsten complexes with a turnover number of 210. In the presence of I, various kinds of electron-deficient alkenes with acetate, ether, carbonyl, and chloro groups at the allylic positions could chemoselectively be oxidized to the corresponding epoxides in high yields with only an equimolar amount of H(2)O(2) with respect to the substrates. Even acrylonitrile and methacrylonitrile could be epoxidized without formation of the corresponding amides. In addition, I could rapidly (<=10 min) catalyze epoxidation of various kinds of terminal, internal, and cyclic alkenes with H(2)O(2) under the stoichiometric conditions. The mechanistic, spectroscopic, and kinetic studies showed that the I-catalyzed epoxidation consists of the following three steps: 1) The reaction of I with H(2)O(2) leads to reversible formation of a hydroperoxo species [gamma-PW(10)O(38)V(2)(MU-OH)(MU-OOH)](3-) (II), 2) the successive dehydration of II forms an active oxygen species with a peroxo group [gamma-PW(10)O(38)V(2)(MU-eta(2):eta(2)-O(2))](3-) (III), and 3) III reacts with alkene to form the corresponding epoxide. The kinetic studies showed that the present epoxidation proceeds via III. Catalytic activities of divanadium substituted polyoxotungstates for epoxidation with H(2)O(2) were dependent on the different kinds of the heteroatoms (i.e., Si or P) in the catalyst and I was more active than [gamma-SiW(10)O(38)V(2)(MU-OH)(2)](4-). On the basis of the kinetic, spectroscopic, and computational results, including those of [gamma SiW(10)O(38)V(2)(MU-OH)(2)](4-), the acidity of the hydroperoxo species in II would play an important role in the dehydration reactivity (i.e., k(3)). The largest k(3) value of I leads to a significant increase in the catalytic activity of I under the more concentrated conditions. PMID- 21647995 TI - Self-organizing domino-like superlattices through stereochemical recognition match at the organic-inorganic interface in solution. PMID- 21647996 TI - Axial preferences in allylations via the Zimmerman-Traxler transition state. PMID- 21647997 TI - Polygoni Rhizoma inhibits inflammatory response through inactivation of nuclear factor-kappaB and mitogen activated protein kinase signaling pathways in RAW264.7 mouse macrophage cells. AB - The objective of this study was to determine the antiinflammatory effects of Polygoni Rhizoma (PR), an Oriental medicinal herb, in interleukin 1 beta (IL 1beta) and lipopolysaccharide (LPS)-stimulated RAW264.7 mouse macrophage cells. PR significantly reduced the production of pro-inflammatory cytokines such as IL 6, tumor necrosis factor alpha (TNF-alpha) and pro-inflammatory mediators such as inducible nitric oxide synthase (iNOS), nitric oxide (NO), cyclooxygenase 2 (COX 2) and prostaglandin E2 (PGE2) even at a concentration of 1 ug/mL in the cells. In addition, PR inhibited the transcriptional activity of NF-kappaB as well as the degradation and phosphorylation of inhibitory kappa B alpha (IkappaBalpha). Furthermore, PR suppressed the phosphorylation of extracellular signal regulated kinase 1/2 (ERK1/2), p38 and c-Jun N-terminal kinase 1/2 (JNK1/2) in IL-1beta and LPS-treated RAW264.7. The results suggest that PR exerts an antiinflammatory property by inhibiting iNOS, COX-2, TNF-alpha and IL-6 production in association with inactivation of the NF-kappaB and MAPK signaling pathways in RAW 264.7 cells. PMID- 21647998 TI - Regulation of obesity and lipid disorders by extracts from Angelica acutiloba root in high-fat diet-induced obese rats. AB - The aim of this study was to investigate the antiobesity and antihyperlipidemic effects of Angelica acutiloba root (Japanese Dong Quai). High-fat diet (HFD) induced obese rats were treated orally with the polyphenolic-rich extract of Angelica acutiloba root (AARE) once daily for 8 weeks. The AARE (300 mg/kg per day) supplementation significantly lowered body weight gain, visceral fat-pad weights and plasma lipid levels, as well as the coronary artery risk index and the atherogenic index of HFD-fed rats. The AARE caused dose related reductions in the hepatic triglyceride and cholesterol contents, as well as lowered hepatic lipid droplet accumulation and epididymal adipocyte size in the HFD-fed rats. The AARE reversed the HFD-induced down-regulation of the hepatic peroxisome proliferator activated receptor-alpha (PPARalpha). The HFD-induced decreases of the hepatic protein level of acyl-CoA oxidase (ACO), and the cytochrome P450 isoform 4A1 (CYP4A1) was up-regulated by AARE. The elevated expressions of hepatic sterol regulatory element binding proteins (SREBPs) of HFD-fed rats were lowered by AARE. These results suggest that AARE attenuated visceral fat accumulation and improved hyperlipidemia in HFD-induced obesity by increasing lipid metabolism through the down-regulation of SREBPs and enhanced the expression of ACO and CYP4A1 in the liver, which was likely mediated by up regulation of the expression of hepatic PPARalpha. PMID- 21647999 TI - Schizencephaly prevalence, prenatal diagnosis and clues to etiology: a register based study. AB - OBJECTIVES: To establish the prevalence and antenatal diagnosis of schizencephaly in the UK. METHODS: Data on schizencephaly were extracted from six regional congenital anomaly registers. RESULTS: Thirty-eight cases of schizencephaly were identified in 2 567 165 livebirths and stillbirths, giving a total prevalence of 1.48/100 000 births (95% CI, 1.01-1.95). Eighteen (47% (95% CI, 31-63%)) of the 38 cases were identified antenatally. No affected fetus had an abnormal karyotype identified. A high proportion of cases of schizencephaly occurred in younger mothers: 63% were aged 24 years or less, significantly higher (P < 0.0001) than the corresponding proportion (26%) of mothers in England and Wales. The majority of cases were not identified until after 22 weeks of pregnancy. Additional anomalies associated with vascular disruption sequences were found in eight cases which had septo-optic dysplasia or absent septum pellucidum, one of which also had gastroschisis. CONCLUSIONS: Schizencephaly occurs more frequently in the fetuses of younger mothers. It is often associated with septo-optic dysplasia, suggesting that the two conditions may share a common origin, arising as a result of destructive processes that cause changes in the brain which only become apparent on ultrasound in the second half of pregnancy. PMID- 21648000 TI - The role of calcium ions in the stability and instability of a thermolysin-like protease. AB - Thermolysin and other secreted broad-specificity proteases, such as subtilisin or alpha-lytic protease, are produced as pre-pro-proteins that stay at least partially unfolded while in the cytosol. After secretion, the pro-proteases fold to their active conformations in a process that includes the autolytic removal of the pro-peptide. We review the life cycle of the thermolysin-like protease from Bacillus stearothermophilus in light of the calcium dependent stability and instability of the N-terminal domain. The protease binds calcium ions in the regions that are involved in the autolytic maturation process. It is generally assumed that the calcium ions contribute to the extreme stability of the protease, but experimental evidence for TLP-ste indicates that at least one of the calcium ions plays a regulatory role. We hypothesize that this calcium ion plays an important role as a switch that modulates the protease between stable and unstable states as appropriate to the biological need. PMID- 21648001 TI - Intermolecular versus intramolecular interactions of the vinculin binding site 33 of talin. AB - The cytoskeletal proteins talin and vinculin are localized at cell-matrix junctions and are key regulators of cell signaling, adhesion, and migration. Talin couples integrins via its FERM domain to F-actin and is an important regulator of integrin activation and clustering. The 220 kDa talin rod domain comprises several four- and five-helix bundles that harbor amphipathic alpha helical vinculin binding sites (VBSs). In its inactive state, the hydrophobic VBS residues involved in binding to vinculin are buried within these helix bundles, and the mechanical force emanating from bound integrin receptors is thought necessary for their release and binding to vinculin. The crystal structure of a four-helix bundle of talin that harbors one of these VBSs, coined VBS33, was recently determined. Here we report the crystal structure of VBS33 in complex with vinculin at 2 A resolution. Notably, comparison of the apo and vinculin bound structures shows that intermolecular interactions of the VBS33 alpha-helix with vinculin are more extensive than the intramolecular interactions of the VBS33 within the talin four-helix bundle. PMID- 21648002 TI - Remarkable change in fluorescence emission of poly(diphenylacetylene) film via in situ desilylation reaction: correlation with variations in microporous structure, chain conformation, and lamellar layer distance. AB - Fluorescence (FL) emission properties, microporous structures, energy-minimized chain conformations, and lamellar layer structures of the silicon-containing poly(diphenylacetylene) derivative of p-PTMSDPA before and after desilylation were investigated. The nitrogen-adsorption isotherms of p-PTMSDPA film before and after desilylation were typical of type I, indicating microporous structures. The BET surface area and pore volume of the p-PTMSDPA film were significantly reduced after the desilylation reaction, simultaneously, its FL emission intensity remarkably decreased. The theoretical calculation on both model compounds of p PTMSDPA and its desilylated polymer, PDPA, showed a remarkable difference in chain conformation: The side phenyl rings of p-PTMSDPA are discontinuously arranged in a zig-zag pattern, while the PDPA is continuously coiled in a helical manner. The lamellar layer distance (LLD) in the p-PTMSDPA film significantly decreased after the desilylation reaction. PMID- 21648003 TI - Synthesis and polymerization of renewable 1,3-cyclohexadiene using metathesis, isomerization, and cascade reactions with late-metal catalysts. AB - Synthesis and subsequent polymerization of renewable 1,3-cyclohexadiene (1,3-CHD) from plant oils is reported via metathesis and isomerization reactions. The metathesis reaction required no plant oil purification, minimal catalyst loading, no organic solvents, and simple product recovery by distillation. After treating soybean oil with a ruthenium metathesis catalyst, the resulting 1,4 cyclohexadiene (1,4-CHD) was isomerized with RuHCl(CO)(PPh3)3. The isomerization reaction was conducted for 1 h in neat 1,4-CHD with [1,4-CHD]/[RuHCl(CO)(PPh3)3] ratios as high as 5000. The isomerization and subsequent polymerization of the renewable 1,3-CHD was examined as a two-step sequence and as a one-step cascade reaction. The polymerization was catalyzed with nickel(II)acetylacetonate/methaluminoxane in neat monomer, hydrogenated d limonene, and toluene. The resulting polymers were characterized by FTIR, DSC, and TGA. PMID- 21648004 TI - Temperature- and redox-directed multiple self assembly of poly(N isopropylacrylamide) grafted dextran nanogels. AB - Poly(N-isopropylacrylamide) (PNIPAAm) grafted dextran nanogels with dodecyl and thiol end groups have been synthesized by RAFT process. Dodecyl-terminated polymers (DexPNI) can be readily dissolved in water and further self assemble into ordered stable nanostructures through direct noncovalent interactions at room temperature. SEM, AFM and DLS measurements confirm the formation of spherical nanogels at hundred-nanometer scales. The elevation of environment temperature will indirectly result in the formation of collapsed nanostructures due to the LCST phase transition of PNIPAAm side chains. Turbidimetry results show that the phase transition behaviors of DexPNI are greatly dependent on PNIPAAm chain length and polymer concentration: increasing PNIPAAm chain length and polymer concentration both lead to lower LCSTs and sharper phase transitions. Moreover, the dodecyl-terminated polymers can transform into thiol-terminated versions by aminolysis of trithiocarbonate groups, and further into chemical (disulfide) cross-linked versions (SS-DexPNI) by oxidation. SS-DexPNI nanogels have "doubled" chain length of PNIPAAm, and hence sharper phase transitions. In situ DLS measurements of the evolution of hydrodynamic radius attest that the self assembly of SS-DexPNI nanogels can be selectively directed by the change in either external temperature or redox potential. These nanogels thus are promising candidates for triggered intracellular delivery of encapsulated cargo. We can also expect that the polymer can be noncovalently (by dodecyl end groups) or covalently (by thiol end groups) coated on a series of nanomaterials (e.g., carbon nanotubes, graphene, gold nanomaterials) to build a variety of novel smart, and robust nanomaterials. PMID- 21648005 TI - Design of bimodal PCL and PCL-HA nanocomposite scaffolds by two step depressurization during solid-state supercritical CO(2) foaming. AB - This communication reports the design and fabrication of porous scaffolds of poly(epsilon-caprolactone) (PCL) and PCL loaded with hydroxyapatite (HA) nanoparticles with bimodal pore size distributions by a two step depressurization solid-state supercritical CO(2) (scCO(2) ) foaming process. Results show that the pore structure features of the scaffolds are strongly affected by the thermal history of the starting polymeric materials and by the depressurization profile. In particular, PCL and PCL-HA nanocomposite scaffolds with bimodal and uniform pore size distributions are fabricated by quenching molten samples in liquid N(2) , solubilizing the scCO(2) at 37 degrees C and 20 MPa, and further releasing the blowing agent in two steps: (1) from 20 to 10 MPa at a slow depressurization rate, and (2) from 10 MPa to the ambient pressure at a fast depressurization rate. The biocompatibility of the bimodal scaffolds is finally evaluated by the in vitro culture of human mesenchymal stem cells (MSCs), in order to assess their potential for tissue engineering applications. PMID- 21648006 TI - "Click" synthesis of fatty acid derivatives as fast-degrading polyanhydride precursors. AB - Fast-degrading linear and branched polyanhydrides are obtained by melt condensation of novel di- and tri-carboxylic acid monomers based on oleic and undecylenic acid synthesized using photoinitiated thiol-ene click chemistry. (1)H NMR spectroscopy, size exclusion chromatography, differential scanning calorimetry, thermogravimetric analysis, and FT-IR spectroscopy have been used to fully characterize these polymers. The hydrolytic degradation of these polymers was studied by means of weight loss, anhydride bond loss, and changes in molecular weight, showing fast degrading properties. Drug release studies from the synthesized polyanhydrides have also been conducted, using rhodamine B as a hydrophobic model drug, to evaluate the potential of these polymers in biomedical applications. PMID- 21648007 TI - Low temperature aqueous living/controlled (RAFT) polymerization of carboxybetaine methacrylamide up to high molecular weights. AB - Among the class of zwitterionic polymers poly(carboxybetaine)s (poly(CB)s) are unique, emerging as the only ultra-low fouling materials known allowing the preparation of biosensors, fouling resistant nanoparticles, and non-adhesive surfaces for bacteria. Poly(carboxybetaine methacrylate) and poly(carboxybetaine acrylamide) have been prepared via atom transfer radical polymerization (ATRP), however a polymerization with living characteristics has not been achieved yet. Herein, the first successful living/controlled reversible addition fragmentation transfer (RAFT) polymerization of (3-methacryloylamino-propyl)-(2-carboxy-ethyl) dimethyl-ammonium (carboxybetaine methacrylamide) (CBMAA-3) in acetate buffer (pH 5.2) at 70 and 37 degrees C is reported. The polymerization afforded very high molecular weight polymers (determined by absolute size exclusion chromatography, close to 250,000 g.mol(-1) in less than 6 h) with low PDI (<1.3) at 70 degrees C. The polymerization was additionally carried out at 37 degrees C allowing to achieve yet lower PDIs (1.06 <= PDI <= 1.15) even at 90% conversion, demonstrating the suitability of the polymerization conditions for bioconjugate grafting. The living character of the polymerization is additionally evidenced by chain extending poly(CBMAA-3) at 70 and 37 degrees C. Block copolymerization from biologically relevant poly[N-(2-hydroxypropyl)methacrylamide] macroCTAs was additionally performed. PMID- 21648008 TI - Frontal polymerization: polymerization induced destabilization of peracrylates. AB - Frontal photopolymerization has attracted much attention in the last decade as it allows the curing of thick films. Unfortunately, the use of peroxides, which feature appropriate storage stability, also requires inappropriately high initiation temperatures. Here, a new approach involving a copolymerisation induced destabilization of (meth)acrylate-based peroxides that allows lower front temperatures is presented. The increasing degree of branching next to the carbonyl group lowers the decomposition temperature by at least 20 degrees C. In classical monomer formulations, sufficient storage stability is confirmed. PMID- 21648009 TI - Continuous production of functionalized polymer particles employing the phase separation in polymer blend films. AB - This study reports a continuous prepartion of spherical or hemispherical polymer particles simply utilizing the phase separation in polymer blend films during the coating process. We took an advantage of the strong phase separation between a water-soluble crystalline polymer as a matrix and hydrophobic polymers as minor components. We demonstrated the prepartion of water-soluble polystyrene (PS) particles, nitrilotriacetic acid (NTA)-functionalized PS particles for protein separation, and semiconducting poly(3-hexylthiophene) (P3HT) particles. The sizes of the particles could be controlled by adjusting the film thickness and weight fraction of the minor component polymers in the blend film. It provides a simple facile way to prepare polymer particles in a continous process. PMID- 21648010 TI - Step-growth radical addition-coupling polymerization (RACP) for synthesis of alternating copolymers. AB - We report a new type of step-growth radical addition-coupling polymerization (RACP) involving consecutive addition of carbon-centered radical derived from alpha,alpha'-dibromo dibasic ester to N?O double bond of C-nitroso compound followed by cross-coupling of carbon-centered radical and in situ formed nitroxyl radical, which produces alternating copolymers with high molecular weight and unimodal molecular weight distribution from saturated and unsaturated monomers. PMID- 21648011 TI - The serum leptin level and body mass index in Melanesian and Micronesian Solomon Islanders: focus on genetic factors and urbanization. AB - OBJECTIVES: This study examined the association between the serum leptin level and body mass index (BMI) and the effects of urbanization and polymorphisms of leptin (LEP) or leptin receptor (LEPR) genes on the leptin level in three Solomon Islands populations. METHODS: A Melanesian population living in a remote area (participants: 106 males and 106 females, ages: 18-74 years), a Melanesian population in an urban area (89 and 94, 18-79 years), and a Micronesian population who migrated to a peri-urban area in the 1960s (84 and 69, 18-71 years) were studied. Anthropometric and serum leptin measurements and genotyping for LEP G-2548A and LEPR K109R and Q223R were performed. RESULTS: The prevalence of obesity (BMI >= 30 kg/m(2)) was the highest in the Micronesian population (30.1%), followed by the urban (18.6%) and the rural (2.4%) Melanesian population. The serum leptin concentration was the highest in the urban Melanesian, followed by the Micronesian and the rural Melanesian populations (P < 0.05). Interestingly, the parameter coefficients of the leptin concentrations on the BMIs were nearly identical in the urban and rural Melanesians after adjusting for age and gender. The LEPR 223Q/Q genotype was associated with an increased leptin level only in the Micronesian population after adjusting for BMI (P = 0.0008 and 0.0016 referenced to the Q/R and the R/R types, respectively). CONCLUSIONS: These observations suggest that the increase in obesity in the Micronesians had a genetic component while that in Melanesians might have been related with the urbanization. PMID- 21648012 TI - Influence of the use of functional foods enriched with phytosterols/-stanols on adherence to statin therapy. AB - PURPOSE: Subjects using functional foods with approved health claims may be more likely to be non-adherent with prescribed drug therapy. This study aimed to assess the influence of the use of phytosterol/-stanol-enriched functional foods on adherence to statin therapy among patients initiating treatment. METHODS: We used data from the statin intervention research project, a randomized controlled trial aimed at improving adherence to statins. In the trial, new statin users were randomized to receive either usual care or extensive pharmaceutical care consisting of five individual counseling sessions. Customary use of phytosterol/ stanol-enriched products was identified by questionnaires filled out by all participants. Automated pharmacy-dispensing records were used to assess adherence in terms of discontinuation of therapy and the medication possession ratio. Analyses were performed for the overall population, as well as stratified for receiving pharmaceutical or usual care. RESULTS: The use of functional foods enriched with phytosterols/-stanols was not related to discontinuation of statin therapy, neither in the overall population (overall population adjusted hazard rate ratio (HR(adj)): 0.80 [95%CI: 0.59-1.08]), nor when stratified by randomization arm (pharmaceutical care HR(adj): 0.77 [95%CI: 0.49-1.23]); usual care HR(adj): 0.81 [95%CI: 0.54-1.21]). The median medication possession ratio was significantly lower in users of phytosterols/-stanols in the usual care group, but the difference was not clinically relevant. CONCLUSIONS: Customary use of phytosterol/-stanol-enriched functional foods did not affect adherence to statins in new users that are well informed on the beneficial effects of statin therapy. In daily medical practice, general practitioners and pharmacists should urge subjects not to take phytosterol/-stanol-enriched functional foods as replacement for their prescribed medication. PMID- 21648013 TI - Ownership and financial sustainability of German acute care hospitals. AB - This paper considers the role of ownership form for the financial sustainability of German acute care hospitals over time. We measure financial sustainability by a hospital-specific yearly probability of default (PD) trying to mirror the ability of hospitals to survive in the market in the long run. The results show that private ownership is associated with significantly lower PDs than public ownership. Moreover, path dependence in the PD is substantial but far from 100%, indicating a large number of improvements and deteriorations in financial sustainability over time. Yet, the general public hospitals have the highest path dependence. Overall, this indicates that public hospitals, which are in a poor financial standing, remain in that state or even deteriorate over time, which may be conflicting with financial sustainability. PMID- 21648014 TI - Estimating adverse selection and moral hazard effects with hospital invoices data in a government-controlled healthcare system. AB - We use invoices for hospital services from a regional hospital in Croatia to test for adverse selection and moral hazard. There are three categories of patients: with no supplemental insurance, who bought it, and who are entitled to it for free. Our identification procedure relies on the premise that the difference in the observed medical care consumption between the patients who bought the insurance and those entitled to free insurance is caused by pure selection effect, whereas the difference in healthcare consumption between the group that received the free insurance and the group that has no insurance is due to moral hazard. Results show favorable selection for patients in 20- to 30-year-old cohort and significant moral hazard for all age cohorts. The selection effect reverses its sign in older cohorts explained by the differences in risk aversion across cohorts caused by the timing of transition from socialism to market economy. PMID- 21648015 TI - Preparation for the end of life in patients with advanced cancer and association with communication with professional caregivers. AB - OBJECTIVE: Previous studies regarding patients' end of life (EOL) preparation have focused mainly on practical tasks, such as advance directives. In this study, we investigate the relational and personal aspects of EOL preparation, using a patient-completed questionnaire, and examine associations with clinician patient communication (CPC) and other variables. METHODS: Patients with advanced cancer but with good performance status were recruited from 24 medical oncology clinics, to participate in a cluster-randomised controlled trial of early palliative care intervention. Measures included the Quality of Life at the End of Life preparation for EOL subscale, and measures of CPC, functional status, comorbidity, spiritual well-being and symptom severity. Using chi-squared tests, t-tests and multivariate regression analyses, we examined the variables associated with preparation for EOL. We also examined the frequency distributions of individual EOL preparation items and used logistic regression to examine their associations with adequacy of CPC. RESULTS: In the 469 patients, characteristics associated with better EOL preparation were better CPC, older age, living alone, less symptom burden and better spiritual well-being. Thirty-one per cent agreed that they worried 'quite a bit' or 'completely' about their family's preparation to cope with the future, and 27% agreed that they would be a burden to their family. All preparation items except regrets about life were associated with adequacy of communication. CONCLUSIONS: A substantial minority of patients with advanced cancer but with good performance status are concerned about EOL preparation, particularly in relation to their families. Better CPC may help patients prepare not only practically but also personally and socially in relation to the dying process and the welfare of their families. PMID- 21648016 TI - Understanding sleep disturbances in African-American breast cancer survivors: a pilot study. AB - OBJECTIVES: The goals of this study were (i) to report the prevalence and nature of sleep disturbances, as determined by clinically significant insomnia symptoms, in a sample of African-American breast cancer survivors; (ii) to assess the extent to which intrusive thoughts about breast cancer and fear of recurrence contributes to insomnia symptoms; and (iii) to assess the extent to which insomnia symptoms contribute to fatigue. METHODS: African-American breast cancer survivors completed surveys pertaining to demographics, medical history, insomnia symptoms, and intrusive thoughts about breast cancer, fear of cancer recurrence, and fatigue. Hierarchical regression models were performed to investigate the degree to which intrusive thoughts and concerns of cancer recurrence accounted for the severity of insomnia symptoms and insomnia symptom severity's association with fatigue. RESULTS: Forty-three percent of the sample was classified as having clinically significant sleep disturbances. The most commonly identified sleep complaints among participants were sleep maintenance, dissatisfaction with sleep, difficulty falling asleep, and early morning awakenings. Intrusive thoughts about breast cancer were a significant predictor of insomnia symptoms accounting for 12% of the variance in insomnia symptom severity. After adjusting for covariates, it was found that insomnia symptom severity was independently associated with fatigue accounting for 8% of variance. CONCLUSIONS: A moderate proportion of African-American breast cancer survivors reported significant problems with sleep. Sleep disturbance was influenced by intrusive thoughts about breast cancer, and fatigue was associated with the severity of participants' insomnia symptoms. This study provides new information about sleep-related issues in African-American breast cancer survivors. PMID- 21648018 TI - Macrophage and microglia ontogeny in the mouse visual system can be traced by the expression of Cathepsins B and D. AB - Here, we show a detailed chronotopographical analysis of cathepsin B and D expression during development of the mouse visual system. Both proteases were detected in large rounded/ameboid cells usually located in close relationship with prominent sites of extensive physiological cell death. In concordance with their morphological features and topographical distribution, we demonstrate that expressing cells corresponded with macrophages and microglial precursors. We found that as microglial precursors differentiated the expression of both cathepsins was down-regulated. Of interest, cathepsin B and D transcripts were never observed in degenerating cells. Our findings point to a role for cathepsin D and B in cell debris degradation after apoptotic processes rather than promoting cell death, as proposed for other developmental models. Additionally their pattern of expression suggests a role in the maturation of the microglial precursors. PMID- 21648017 TI - Gene expression profile of the regeneration epithelium during axolotl limb regeneration. AB - Urodele amphibians are unique among adult vertebrates in their ability to regenerate missing limbs. The process of limb regeneration requires several key tissues including a regeneration-competent wound epidermis called the regeneration epithelium (RE). We used microarray analysis to profile gene expression of the RE in the axolotl, a Mexican salamander. A list of 125 genes and expressed sequence tags (ESTs) showed a >=1.5-fold expression in the RE than in a wound epidermis covering a lateral cuff wound. A subset of the RE ESTs and genes were further characterized for expression level changes over the time course of regeneration. This study provides the first large scale identification of specific gene expression in the RE. PMID- 21648019 TI - Concise review: challenging the pluripotency of human testis-derived ESC-like cells. AB - Several research groups have claimed to have successfully generated pluripotent or multipotent cells from human testis. However, the pluripotent character of those cells with respect to gene expression profile and ability to generate teratomas has been called into question. Here, we critically review these reports and provide insight to guide future studies on the derivation of human pluripotent cells from testicular tissue. PMID- 21648020 TI - Haplotype-based analysis of ulcerative colitis risk loci identifies both IL2 and IL21 as susceptibility genes in Han Chinese. AB - BACKGROUND: The incidence of ulcerative colitis (UC) varies between Western and Eastern ethnicities. A distinct genetic background may play a role in the differences. Until now, very little was known of the UC genetics in Asian populations. Here we performed a haplotype-based analysis of six known UC susceptibility loci in Han Chinese patients. METHODS: In all, 245 UC patients and 300 healthy controls of Han Chinese descent were genotyped for 27 single nucleotide polymorphisms (SNPs), which cover the major haplotypes of the chromosome regions containing IL10, IL2/IL21, MYO9B, ECM1, MST1, and IL23R in Han Chinese. RESULTS: In contrast to the tight linkage disequilibrium (LD) block of the IL2/IL21 region in Caucasians, IL2 and IL21 reside in two independent LD blocks in Han Chinese. The IL2 SNP rs2069762 (P = 7.0 * 10(-4) , odds ratio [OR] = 1.54, 95% confidence interval [CI] 1.20-1.99) and the IL21 SNP rs2055979 (P = 1.2 * 10(-4) , OR = 1.50, 95% CI 1.17-1.92) were independently associated with UC. We identified one risk haplotype in IL2 and another independent risk haplotype in IL21. In addition to the IL2/IL21 locus, we observed association of the TT genotype of SNP rs1545620 in MYO9B with UC (P = 0.0169; OR = 0.29, 95% CI 0.11-0.78) and association of rs17375018 in IL23R with pancolitis in Chinese UC patients (P = 0.002; OR = 2.38, 95% CI 1.41-4.02). CONCLUSIONS: Our study confirmed the association of the IL2/IL21 region with UC in Han Chinese patients, and further implied both IL2 and IL21 as genetic risk factors for UC. Han Chinese UC patients share part of their genetic susceptibility with Caucasian patients. PMID- 21648021 TI - Cancer prevention in ulcerative colitis: long-term outcome following fluorescence guided colonoscopy. AB - BACKGROUND: Patients with long-standing ulcerative colitis require repeated endoscopies for early detection of neoplasias, which, however, are frequently missed by standard colonoscopy. Fluorescence-guided colonoscopy is known to improve the detection rate but the long-term effects of fluorescence-guided colonoscopy are unknown. METHODS: Colitis patients with negative findings at index fluorescence-guided colonoscopy entered a prospective long-term study with conventional colonoscopies at 2-year intervals. Risk and time to progression were evaluated. The positive predictive value was assessed in patients with neoplasias at index fluorescence-guided colonoscopy who underwent immediate total colectomy. RESULTS: Thirty-one patients with negative fluorescence-guided colonoscopy were surveyed for a mean of 7.8 +/- 0.9 years. Neoplasia was observed in only two of them (6%) after 7 and 8 years of follow-up, respectively. Neoplasia at index fluorescence-guided colonoscopy was observed in 10 patients. In all of them, multiple flat low-grade intraepithelial neoplasia was diagnosed. At immediate colectomy performed in eight of them, the diagnosis of flat low-grade intraepithelial neoplasia was confirmed, corresponding to a positive predictive value of 100%. However, synchronous more advanced neoplasia was detected in three of the eight patients (38%). All patients, those with and those without neoplasia, were alive at the end of the study. CONCLUSIONS: Fluorescence-guided colonoscopy misses, in contrast to standard colonoscopy, few, if any, patients with neoplasia. Most neoplasia-negative patients remain negative during prolonged follow-up. However, when low-grade dysplasia is diagnosed by fluorescence-guided colonoscopy, colectomy is recommended because more than a third of the patients harbor synchronous, more advanced neoplasia. PMID- 21648022 TI - Abdominal phlegmons in Crohn's disease: outcomes following antitumor necrosis factor therapy. AB - BACKGROUND: An abdominal phlegmon is an inflammatory mass that can develop in the setting of penetrating Crohn's disease (CD). Anti-tumor necrosis factor (TNF) antibody therapy is typically avoided in CD complicated by phlegmon because of concern for peritoneal infection but may offer an effective alternative to surgical resection after infection has been controlled with antibiotics. The aim of this study was to examine outcomes for patients with CD who developed an abdominal phlegmon that was subsequently treated with an anti-TNF antibody. METHODS: We retrospectively reviewed the records of all CD patients attending Beth Israel Deaconess Medical Center between 2004 and 2010 with an abdominal phlegmon who were treated with an anti-TNF antibody in order to evaluate the safety and efficacy of this treatment regimen. RESULTS: There were 13 patients with CD complicated by a phlegmon treated with antibiotics and an anti-TNF antibody at our center between 2004 and 2010. Ten were male. Median time (interquartile range) from diagnosis to development of the phlegmon was 5.9 (1.9 22.7) years. The phlegmon was associated with an abscess in 12 patients. In addition to anti-TNF therapy, all patients were treated with broad-spectrum antibiotics. Anti-TNF therapy was effective without complications in all subjects. Two patients eventually had surgery more than a year after initiating anti-TNF treatment. CONCLUSIONS: Penetrating CD complicated by phlegmon formation may be safely and effectively managed with a combination of antibiotics and anti TNF therapy. Larger, prospective trials are required to confirm these initial findings. PMID- 21648023 TI - Human herpes virus-6 chromosomal integration misled the management of Crohn's disease. PMID- 21648024 TI - Hormones, heart disease, and health: individualized medicine versus throwing the baby out with the bathwater. AB - It is increasingly axiomatic that depression has widespread adverse physiological effects and, conversely, that a variety of physiological systems impact the risk for developing depression. This convergence of depression and altered physiology is particularly dramatic during midlife--a time during which reproductive failure presages dramatic increases in prevalence of both heart disease and depression. The potentially meaningful and illuminating links between estrogen deficiency, cardiovascular disease (CVD), and depression have largely been obscured, first by assertions, subsequently repudiated, that the perimenopause was not a time of increased risk of depression, and more recently by the denegration of hormone replacement therapy by initial reports of the Women's Health Initiative. Increasingly, however, research has led to unavoidable conclusions that CVD and depression share common, mediating pathogenic processes and that these same processes are dramatically altered by the presence or absence of estrogen (E2). This review summarizes data supporting these contentions with the intent of placing depression and estrogen therapy in their proper physiologic context. PMID- 21648027 TI - Selective reduction of carboxylic acid derivatives by catalytic hydrosilylation. AB - In the last decade, an increasing number of useful catalytic reductions of carboxylic acid derivatives with hydrosilanes have been developed. Notably, the combination of an appropriate silane and catalyst enables unprecedented chemoselectivity that is not possible with traditional organometallic hydrides or hydrogenation catalysts. For example, amides and esters can be reduced preferentially in the presence of ketones or even aldehydes. We believe that catalytic hydrosilylations will be used more often in the future in challenging organic syntheses, as the reaction procedures are straightforward, and the reactivity of the silane can be fine-tuned. So far, the synthetic potential of these processes has clearly been underestimated. They even hold promise for industrial applications, as inexpensive and readily available silanes, such as polymethylhydrosiloxane, offer useful possibilities on a larger scale. PMID- 21648028 TI - Fast multidimensional NMR spectroscopy using compressed sensing. PMID- 21648029 TI - Synthesis of gold nano-hexapods with controllable arm lengths and their tunable optical properties. PMID- 21648030 TI - Protein chemical synthesis by ligation of peptide hydrazides. PMID- 21648031 TI - Superoleophobic coatings with ultralow sliding angles based on silicone nanofilaments. PMID- 21648032 TI - Synthesis of anionic iron polyphosphides by reaction of white phosphorus with "Cp*Fe-". PMID- 21648033 TI - Asymmetric binary acid catalysis: a regioselectivity switch between enantioselective 1,2- and 1,4-addition through different counteranions of In(III). PMID- 21648034 TI - Asymmetrically functionalized graphene for photodependent diode rectifying behavior. PMID- 21648035 TI - A metal-free "threading-followed-by-shrinking" protocol for rotaxane synthesis. PMID- 21648036 TI - A potent trivalent sialic acid inhibitor of adenovirus type 37 infection of human corneal cells. PMID- 21648037 TI - Separation of metallic and semiconducting single-walled carbon nanotube arrays by "scotch tape". PMID- 21648038 TI - Snapshots of complete nitrogen atom transfer from an iron(IV) nitrido complex. PMID- 21648039 TI - Pure alpha-metallated benzyllithium from a single-crystal-to-single-crystal transition. PMID- 21648040 TI - Dopamine as a carbon source: the controlled synthesis of hollow carbon spheres and yolk-structured carbon nanocomposites. PMID- 21648041 TI - La/Ag heterobimetallic cooperative catalysis: a catalytic asymmetric Conia-ene reaction. PMID- 21648042 TI - Conversion of cobalt(II) porphyrin into a helical cobalt(III) complex of acyclic pentapyrrole. PMID- 21648043 TI - Giant negative thermal expansion in the iron perovskite SrCu3Fe4O12. PMID- 21648044 TI - Gold-catalyzed oxidative cyclization of 1,5-enynes using external oxidants. PMID- 21648045 TI - Spectroscopy of dibenzorubicene: experimental data for a search in interstellar spectra. AB - We report on the characterization of dibenzo[cde,opq]rubicene (C(30)H(14)). The molecule was studied in solution at room temperature with absorption spectroscopy in the visible (vis) and ultraviolet (UV) wavelength ranges, and with emission spectroscopy. The infrared (IR), visible, ultraviolet, and vacuum ultraviolet (VUV) absorption spectra of a thin film were measured also at room temperature. In addition, the UV/vis absorption spectrum was measured at cryogenic temperatures using the matrix isolation spectroscopy technique. The interpretation of spectra was supported by theoretical calculations based on semiempirical and ab initio models, as well as on density functional theory. Finally, the results of the laboratory study were compared with interstellar spectra. PMID- 21648046 TI - Experimental and theoretical investigation of vibrational spectra of coordination polymers based on TCE-TTF. AB - The room-temperature infrared and Raman spectra of a series of four isostructural polymeric salts of 2,3,6,7-tetrakis(2-cyanoethylthio)-tetrathiafulvalene (TCE TTF) with paramagnetic (Co(II), Mn(II)) and diamagnetic (Zn(II), Cd(II)) ions, together with BF(4)(-) or ClO(4)(-) anions are reported. Infrared and Raman active modes are identified and assigned based on theoretical calculations for neutral and ionized TCE-TTF using density functional theory (DFT) methods. It is confirmed that the TCE-TTF molecules in all the materials investigated are fully ionized and interact in the crystal structure through cyanoethylthio groups. The vibrational modes related to the C=C stretching vibrations of TCE-TTF are analyzed assuming the occurrence of electron-molecular vibration coupling (EMV). The presence of the antisymmetric C=C dimeric mode provides evidence that charge transfer takes place between TCE-TTF molecules belonging to neighboring polymeric networks. PMID- 21648047 TI - Components of sleep quality and sleep fragmentation in rheumatoid arthritis and osteoarthritis. AB - OBJECTIVES: Poor sleep is increasingly recognized as contributing to a decreased quality of life, increased morbidity/mortality and heightened pain perception. The purpose of the present study was to assess components of sleep quality and self-identified contributors to sleep fragmentation in rheumatoid arthritis (RA) and osteoarthritis (OA) patient populations. METHODS: Consecutive RA and OA clinic patients were invited to participate in a self-administered questionnaire study which included the validated multi-domain Pittsburgh Sleep Quality Index (PSQI), visual analogue scales for pain, fatigue, global functioning, modified Health Assessment Questionnaire (mHAQ), stress scores, the Centre for Epidemiologic Studies-Depression (CES-D) score, the 36-item short form (SF-36) quality of life measure, the Rheumatoid Arthritis Disease Activity Index (RADAI), the Epworth Sleepiness Scale (ESS), Berlin score for obstructive sleep apnoea (OSA) risk and the International Restless Legs Syndrome Study Group (IRLSSG) diagnostic criteria. RESULTS: The study population included 145 RA and 78 OA patients. PSQI global scores were >5 in 62% of RA and 67% of OA patients. Multivariate analysis identified global functioning and the CES-D to be independent predictors for higher global PSQI scores in RA patients, whereas in OA patients predictors were the mHAQ and SF-36 mental component summary. Abnormalities in subjective sleep assessment, sleep latency, sleep duration, sleep efficiency, daytime dysfunction and increased sleep-aid medication use were observed in both populations. The most common abnormality reported by both RA and OA patients was increased sleep fragmentation. The most frequent self-identified cause for sleep disturbance was 'need to use the washroom' by 51% of RA and 49% of OA patients, and, second most common, 'pain' was identified as a cause for awakening by 33% of RA and 45% of OA patients. CONCLUSIONS: A high prevalence of abnormal sleep quality in both RA and OA patient populations was observed. The most common abnormality was sleep fragmentation, with an increased sleep disturbance score. 'Need to use the washroom' and 'pain' were the most common self-identified reasons for awakening from sleep. A review of sleep hygiene, optimization of urological status, and rheumatological disease symptomatic control may prove beneficial in terms of sleep health. PMID- 21648048 TI - Treatment of aortocoronary dissection complicating anomalous origin right coronary artery and chronic total intervention with intravascular ultrasound guided stenting. AB - Acute dissection of the ascending aorta is a rare complication of percutaneous coronary intervention (PCI). Its mechanism involves disruption of the coronary intima by mechanical trauma, followed by subintimal injection of contrast, which, in turn, contributes to subsequent extension of the dissection. In contrast to spontaneous aortic dissection of ascending aorta, which mandates immediate surgical intervention, the appropriate therapy and outcome of this rare entity are not well established. We report a case of iatrogenic aortocoronary dissection, complicating transradial PCI for recanalization of anomalous origin right coronary artery (RCA) from the left coronary cusp with chronic total occlusion. The intimal tear was created by a balloon rupture in the proximal RCA, with propagation of dissection into the ostium and the coronary sinus of Valsalva. Intravascular ultrasound (IVUS) guided coronary stenting was performed to seal the entry port, and to break down the dissection route. This case indicates that IVUS can be a useful tool to ensure complete coverage of the intracoronary dissection and precise placement of the stent to fully cover the ostium of the culprit vessel. This can be particularly important in difficult situations, such as anomalous origin of RCA from the left cusp, with acute downward anterior angulation. PMID- 21648049 TI - Very late thrombosis in a bare metal stent: mechanistic analysis from optical coherence tomography. AB - Recent publications have emphasized the concept that restenosis of a bare metal stent (BMS) is not as clinically benign as previously believed. We present a case of very late stent thrombosis 9 years after implantation of a BMS is described. Optical coherence tomography shows plaque rupture in the stent. These findings suggest that the formation and progression of a neoatheroma over a previously healed intrastent vascular intima may represent a "new" mechanism for stent thrombosis. Evidence from the current case supports this hypothesis. PMID- 21648050 TI - Retrograde access via the popliteal artery to facilitate the re-entry technique for recalcitrant superficial femoral artery chronic total occlusions. AB - Subintimal recanalization is beneficial in selected patients with peripheral chronic total occlusions (CTO). However, in complex cases, re-entry into the true arterial lumen may prove to be unsuccessful with a conventional guidewire or a re entry catheter when using standard femoral artery access. Our case series describes these technical dilemmas along with strategies that can be utilized to overcome these challenges. PMID- 21648051 TI - Emergency manual systemic-to-pulmonary artery autotransfusion for severe cyanotic spell. AB - Cyanotic spell is an important complication of tetralogy of Fallot. We report a simple, innovative method that bailed out a patient with life-threatening, refractory cyanotic spell in the catheterization laboratory. Manual autotransfusion of blood from aorta into the pulmonary arteries resulted in significant clinical improvement. PMID- 21648052 TI - Objective performance goals of safety and blood pressure efficacy for clinical trials of renal artery bare metal stents in hypertensive patients with atherosclerotic renal artery stenosis. AB - OBJECTIVE: To provide safety and performance goals for prospective single-arm trials of bare metal renal artery stenting in patients with resistant hypertension associated with high grade atherosclerotic renal artery stenosis. BACKGROUND: To date, there have been no US Pre-Market Approval (PMA) bare metal renal stent device trials which have focused on improvement of blood pressure control as a primary effectiveness endpoint. METHODS: Analysis of subject-level data from three large industry sponsored pre-market approval (PMA) trials was performed. Hypertensive patients (>= 155 mmHg) with a >= 50% atherosclerotic renal artery stenosis were included. Thirty day and 9-month systolic and diastolic blood pressure measurements, renal function and 9-month duplex ultrasound assessment of renal artery patency were analyzed. RESULTS: Initial data analysis of 600 patients from the 3 PMA trials identified 286 patients who met inclusion criteria. The mean baseline systolic blood pressure was 177.8 +/- 19.3 mmHg with a mean 68.1% diameter renal artery stenosis. Nine months after successful stenting, the mean SBP was 156.7 +/- 24.1 mmHg; the 9 month restenosis rate was 14.4%. CONCLUSION: Based on the statistical modeling of these data and a priori established performance criteria, the co-primary endpoints of 9 month reduction in blood pressure and in-stent restenosis are proposed. The reduction in blood pressure will be analyzed as a continuous variable and will be compared to this performance goal. PMID- 21648053 TI - The results of transcatheter closure of VSD using Amplatzer(r) device and Nit Occlud(r) Le coil. AB - OBJECTIVE: We retrospectively reviewed the result of patients who underwent transcatheter closure of ventricular septal defect (VSD) using Amplatzer(r) Perimembranous or Amplatzer(r) muscular VSD device (the Amplatzer(r) group) and Nit Occlud(r) Le VSD Coil (the Pfm group). BACKGROUND: Perimembranous type (PmVSD) and doubly committed subarterial (DCSA) VSD were the major isolated congenital heart defects in Thai children. Transcatheter device closure technique for both types of VSD has emerged as an alternative treatment to surgery. METHODS: Retrospectively, data was reviewed between 2003 and 2009. RESULTS: 116 patients were enrolled. Device or coil was successfully implanted in 94%. Complete closure at 24 hr is slightly higher in the Amplatzer(r) group. The average size of VSD in the Amplatzer(r) group was larger than the Pfm group (P = 0.001). The Pfm coil was primarily deployed in DCSA VSD when compared with the Amplatzer(r) group (P < 0.01). At 6 months follow-up the residual shunt was comparable (P = 0.054). There was only one transient AV block (AVB) in the Pfm group and 5 AVB in the Amplatzer(r) group. Four pacemakers were placed in the Amplatzer(r) group. CONCLUSIONS: Transcatheter closure of VSD in both Pm VSD and DCSA can be achieved by using either of the device. The Amplatzer(r) VSD device had the advantage of closure of larger defects with immediate less residual shunt but appeared to have a significant number of 3 degrees AVB, which required pacemaker implantation. The Nit Occlud(r) Le VSD Coil had the advantage of closure of both types of defects, in particular DCSA VSD with only small residual shunt. PMID- 21648054 TI - Collateral blood flow can predict myocardial blush grade in primary coronary intervention. AB - OBJECTIVES: Our aim was to investigate whether collateral flow may predict myocardial blush grade (MBG) in acute myocardial infarction patients undergoing primary percutaneous coronary intervention. BACKGROUND: No-reflow is a well known phenomenon associated with increased morbidity and mortality due to underperfused myocardium; therefore early prediction of no-reflow is of major importance. We have observed that in patients with good collateral filling of the infarct related artery as seen prior to primary angioplasty, the clearance of the contrast medium from the myocardium may be impaired. METHODS: We retrospectively analyzed the MBG as observed by collateral filling in 81 patients and correlated it with the final MBG. Patients were divided into two groups-those with collateral MBG 0 or 1 (34) and those with myocardial blush 2 or 3 (47). RESULTS: Of the 34 patients in the first group 71% remained in the same MBG group after primary percutaneous coronary intervention and the rest improved. Of the 47 individuals with collateral MBG 2 or 3, 87% remained in the same group following primary percutaneous coronary intervention, and the rest deteriorated (P < 0.01 for both groups). CONCLUSIONS: Collaterals may predict MBG in acute myocardial infarction patients undergoing primary percutaneous coronary intervention. PMID- 21648056 TI - Maternal use of hot tub and major structural birth defects. AB - BACKGROUND: Previous studies on the associations between hot tub use during early pregnancy and birth defects have found an increased risk of neural tube defects, but no increase in risk of cardiac defects. No previous studies have assessed the association between maternal hot tub use and other types of noncardiac birth defects. METHODS: We included mothers of infants with birth defects (n = 10,825) and mothers of infants without birth defects (n = 6795) who participated in the multisite National Birth Defects Prevention Study between 1997 and 2005. Odds ratios were adjusted for maternal ethnicity and education. RESULTS: Analysis of 17 birth defects revealed that mothers of infants with gastroschisis and anencephaly were significantly more likely to report any use of a hot tub in early pregnancy: adjusted odd ratios were 1.54 (95% confidence interval [CI], 1.10-2.17) and 1.68 (95% CI, 1.05-2.70), respectively. Among the mothers who reported using a hot tub more than once in the exposure period and remaining in it for more than 30 min, we found significantly elevated odds ratios (>=2.0) for esophageal atresia, omphalocele, and gastroschisis and a nonsignificant elevation (>=2.0) for spina bifida and anencephaly. CONCLUSIONS: These results suggest that women who use hot tubs more than once during early pregnancy and for long periods of time have an increased risk of certain birth defect phenotypes, particularly anencephaly and gastroschisis. Because of multiple statistical tests and small sample sizes, we cannot exclude the possibility that some of these elevated associations may be due to chance. PMID- 21648055 TI - The role of imprinted genes in fetal growth abnormalities. AB - Epigenetics, and in particular imprinted genes, have a critical role in the development and function of the placenta, which in turn has a central role in the regulation of fetal growth and development. A unique characteristic of imprinted genes is their expression from only one allele, maternal or paternal and dependent on parent of origin. This unique expression pattern may have arisen as a mechanism to control the flow of nutrients from the mother to the fetus, with maternally expressed imprinted genes reducing the flow of resources and paternally expressed genes increasing resources to the fetus. As a result, any epigenetic deregulation affecting this balance can result in fetal growth abnormalities. Imprinting-associated disorders in humans, such as Beckwith Wiedemann and Angelman syndrome, support the role of imprinted genes in fetal growth. Similarly, assisted reproductive technologies in animals have been shown to affect the epigenome of the early embryo and the expression of imprinted genes. Their role in disorders such as intrauterine growth restriction appears to be more complex, in that imprinted gene expression can be seen as both causative and protective of fetal growth restriction. This protective or compensatory effect needs to be explored more fully. PMID- 21648057 TI - Rhabdomyomatous differentiation in primary Wilms tumor and hepatic metastases after chemotherapy and all-trans-retinoic acid in combination with interferon alpha. PMID- 21648058 TI - Portland cement for bone tissue engineering: Effects of processing and metakaolin blends. AB - The need for a suitable scaffolding material for load bearing bone tissue engineering still has yet to be met satisfactorily. In this study, Portland cement and Portland cement/metakaolin (MK) blends were processed to render them biologically and mechanically suitable for such application. Portland cement was mixed with MK at different ratios. The slurries were hydrated under atmospheric (noncarbonated samples) and high-CO2 conditions (carbonated samples). The mechanical properties were characterized via compressive tests. The bioactivity was analyzed in a simulated body fluid solution. Scanning electron microscopy and energy dispersive spectroscopy were used to evaluate sample morphology and chemistry. The cytocompatibility (direct contact assay, MTT test, and alkaline phosphatase activity) was tested using human osteoblast-like cells. Cell responses were observed via conventional and electron microscopy. The results showed that the implementation of MK did not significantly influence the mechanical properties. All the samples evidenced bioactive behavior. Cell experiments confirmed a highly cytotoxic response to the noncarbonated specimens. The introduction of MK as well as the CO2 pretreatment significantly improved the cytocompatibility of the specimens. These results show that properly processed Portland cement and Portland cement/MK blends could present suitable properties for the development of load-bearing scaffolding structures in bone tissue engineering applications. PMID- 21648059 TI - Development of a biodegradable foam for use in negative pressure wound therapy. AB - Treatment of wounds using negative pressure wound therapy (NPWT) uses a nondegradable polyvinyl alcohol (PVA) foam in the application of negative pressures typically for 1-3 days. The purpose of this study was to construct and test biodegradable poly(epsilon-caprolactone) (PCL) foam as a substitute for the PVA foam. Such a foam would be left within the wound until healing was achieved and form a biodegradable matrix into which tissue would grow. The use of such foam would obviate the need for any serial foam changes and a final foam removal, thus making patient care much easier and more economical. PCL foams were prepared by salt leaching and phase separation. Morphological and mechanical properties of the foams were characterized and compared to PVA foam. PCL and PVA foams were tested on the uncut surface of a pig liver maintained in a hydration chamber continuously replenished with saline under the conditions of negative pressure of 50 mm Hg for 72 h. The results demonstrated that PCL foam made from phase separation had the similar properties and function as the PVA foam. The results demonstrate that PCL foam is an appropriate substitute for currently used nondegradable PVA foam in NPWT applications. PMID- 21648060 TI - Complement activation by candidate biomaterials of an implantable microfabricated medical device. AB - Implantable devices realized by microfabrication have introduced a new class of potential biomaterials whose properties would need to be assessed. Such devices include sensors for measuring biological substances like glucose. Thus, 14 different candidate materials intended for design of such a device were investigated with respect to their complement activation potential in human serum. The fluid-phase activation was measured by the products C4d, Bb, C3bc, and the terminal complement complex (TCC), whereas solid-phase activation was measured by deposition of TCC on the material surfaces. No fluid-phase activation was found for materials related to the capsule, carrier, or sealing. Fluid-phase activation was, however, triggered to a various extent in three of the four nanoporous membranes (cellulose, polyamide, and aluminium oxide), whereas polycarbonate was rendered inactive. Solid-phase activation discriminated more sensitively between all the materials, revealing that the capsule candidate polydimethylsiloxane and sealing candidate silicone 3140 were highly compatible, showing significantly lower TCC deposition than the negative control (p < 0.01). Three of the candidate materials were indifferent, whereas the remaining nine showed significantly higher deposition of TCC than the negative control (p < 0.01). In conclusion, complement activation, in particular when examined on the solid phase, discriminated well between the different candidate materials tested and could be used as a guide for the selection of the best-suited materials for further investigation and development of the device. PMID- 21648061 TI - Cell adhesion and proliferation studies on semi-interpenetrating polymeric networks (semi-IPNs) of polyacrylamide and gelatin. AB - In this study, the effect of feed composition, degree of hydrophilicity, and internal morphology has been investigated for cell proliferation potential of the polyacrylamide/gelatin (PAm/G) semi-interpenetrating polymeric network (semi IPNs). Polycaprolactone diacrylate was used to cross-link polyacrylamide chains. Scanning electron microscopy (SEM) micrographs demonstrate uniformly distributed porous structure with internal diameter in the range of 75-175 MUm, dependent on matrix compositions. Water-air contact angle was found in the range of 49 degrees +/- 0.22 to 89 degrees +/- 0.14 (p < 0.02) suggesting varying degree of hydrophilicity of the hydrogel surface. In addition, protein adsorption study showed 45 +/- 0.14 MUg to 64 +/- 0.12 MUg (p < 0.01) of protein adsorbed per cm2 of hydrogel. Quantitative estimation of cell adhesion and proliferation was carried out by DNA quantification using fluorimetric assay method (p < 0.02). Microscopic images of proliferative cells on semi-IPNs by fluorescent and inverted phase contrast supported the findings of DNA quantification. Contact angle in the range of 63-69 degrees in association with 52-59 MUg/cm2 protein absorption and 115-150 MUm pore size was found optimum for fibroblast proliferation on PAm/G semi-IPN scaffolds. The newly developed semi interpenetrating network may serve as a potential scaffold for soft tissue engineering applications. PMID- 21648062 TI - Spatiotemporal control of proliferation and differentiation of bone marrow derived mesenchymal stem cells recruited using collagen hydrogel for repair of articular cartilage defects. AB - Articular cartilage has a poor healing capacity, and cartilage regeneration is not always warranted to achieve healing. On the other hand, collagen scaffolds have been shown to support regeneration of articular cartilage defects in animal models, whereas bone morphogenetic protein-2 (BMP-2) is known to cause chondrogenic differentiation of marrow-derived mesenchymal stem cells (MSCs). The purpose of this study was to evaluate the effectiveness of intra-articular administration of BMP-2 into bone marrow-derived MSCs recruited to defects using original collagen hydrogel in rabbits at various time points. Full-thickness defects were created in both knees, then collagen hydrogels were transplanted, and BMP-2 was supplied for 1-week periods, as follows. BMP-2 was administered immediately after the operation for 1 week (BMP0-1 group), and BMP-2 was administered between weeks 1 and 2 after the operation (BMP1-2 group). BMP2 was administered between weeks 2 and 3 (BMP2-3 group). Specimens were then obtained, and bromodeoxyuridine (BrdU)-positive cells were enumerated and histologic grading was also performed. In addition, the gene expression analysis was performed using quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) assays. Enumeration of BrdU-positive cells showed a significant increase in the BMP0-1 group compared with the other groups. Similarly, histologic scores in the BMP0-1 group were superior for up to 8 weeks. Finally, RT-PCR findings revealed that immediate BMP-2 administration enhanced chondrogenic differentiation. PMID- 21648063 TI - Electrochemical investigation of chromium oxide-coated Ti-6Al-4V and Co-Cr-Mo alloy substrates. AB - Hard coatings for articulating surfaces of total joint replacements may improve the overall wear resistance. However, any coating approach must take account of changes in corrosion behavior. This preliminary assessment analyzes the corrosion kinetics, impedance and mechanical-electrochemical stability of 100 MUm thick plasma sprayed chromium oxide (Cr2O3) coatings on bearing surfaces in comparison to the native alloy oxide films on Co-Cr-Mo and Ti-6Al-6V. Cyclic potentiodynamic polarization, electrochemical impedance spectroscopy, and mechanical abrasion under potentiostatic conditions were performed on coated and substrate surfaces in physiological saline. SEM analysis characterized the coating morphology. The results showed that the corrosion current density values of chromium oxide coatings (0.4-1.2 MUA/cm2) were of the same order of magnitude as Ti-6Al-4V alloy. Mechanical abrasion did not increase corrosion rates of chromium oxide coatings but did for uncoated Co-Cr-Mo and Ti-6Al-4V. The impedance response of chromium oxide coatings was very different than Co-Cr-Mo and Ti-6Al-4V native oxides characterized by a defected coating model. More of a frequency-independent purely resistive response was seen in mid-frequency range for the coatings (CPE(coat) : 40-280 nF/cm2 (rad/s)(1-alpha) , alpha: 0.67-0.83) whereas a more capacitive character is seen for Co-Cr-Mo and Ti-6Al-4V (CPE(ox) around 20 MUF/cm2 (rad/s)(1-alpha) , alpha around 0.9). Pores, interparticle gaps and incomplete fusion typical for thermal spray coatings were present in these oxides which could have influenced corrosion resistance. The coating microstructure could have allowed some fluid penetration. Overall, these coatings appear to have suitable corrosion properties for wear surfaces. PMID- 21648064 TI - Deproteinized dentin: a favorable substrate to self-bonding resin cements? AB - The adhesive performance on deproteinized dentin of different self-adhesive resin cements was evaluated through microtensile bond strength (MUTBS) analysis and scanning electron microscopy (SEM). Occlusal dentin of human molars were distributed into different groups, according to the categories: adhesive cementation with two-step bonding systems-control Groups (Adper Single Bond 2 + RelyX ARC/3M ESPE; One Step Plus + Duolink/Bisco; Excite + Variolink I/Ivoclar Vivadent) and self-adhesive cementation-experimental groups (Rely X Unicem/3M ESPE; Biscem/Bisco; MultiLink Sprint/Ivoclar Vivadent). Each group was subdivided according to the dentin approach to: alpha, maintenance of collagen fibers and beta, deproteinization. The mean values were obtained, and submitted to ANOVA and Tukey test. Statistical differences were obtained to the RelyX Unicem groups (alpha = 13.59 MPa; beta = 30.19 MPa). All the BIS Group specimens failed before the mechanical tests. Dentinal deproteinization provided an improved bond performance for the self-adhesive cement Rely X Unicem, and had no negative effect on the other cementing systems studied. PMID- 21648065 TI - Surface characteristics and biological studies of hydroxyapatite coating by a new method. AB - Hydroxyapatite was coated on the Ti implants by a new coating method called low temperature high speed collision (LTHSC). Higher roughness values were estimated in the formed thick coating on the implant surface. While the roughness value was lower than plasma sprayed HA coatings, LTHSC HA-coated surface showed higher hydrophilicity. The cell proliferation of oseteoblastic cells evaluated by MTT assay showed that HA-coated surface had significantly higher cell viability than the control. The HA-coated surface showed higher mean total protein production even though there was no statistical difference between two surfaces. Osteoblast differentiation, ALP activity and expression of differentiation marker genes such as osteopontin and osteocalcin were elevated in the cells cultured on HA-coated surface and this surface also showed higher collagen protein expression than titanium control. Moreover, in experimental group, BIC values were significantly increased after 12 weeks of implantation and about 30-50% cortical bone-implant contact was observed. PMID- 21648066 TI - The intrinsically high pitting corrosion resistance of mechanically polished nitinol in simulated physiological solutions. AB - Nitinol wires have been widely used in many biomedical applications, such as cardiovascular stent due to their superelasticity and shape memory effect. However, their corrosion properties and the related biocompatibility are not well understood, and the reported results are controversial. In this study, we evaluate the pitting corrosion property of nitinol, titanium, nickel, and 316L stainless steel (316LSS) wires with different surface roughnesses in a saline solution at 37 degrees C. The cyclic potentiodynamic polarization results show that mechanically polished nitinol and Ti wires are highly resistant to pitting corrosion, while Ni and 316LSS wires are susceptible to pitting corrosion. Electrochemical impedance spectroscopy is used to study the interface of oxide film/solution and all mechanically polished nitinol wires are covered by 2-3 nm thick films formed under open circuit potential. Furthermore, the electronic structures and semiconducting properties of passive films on nitinol, Ti and Ni wires are studied by Mott-Schottky analysis. Passive films formed on nitinol and Ti exhibit n-type semiconducting characteristics, whereas films on Ni show p-type semiconducting characteristics. Scanning Kelvin Microscopy is used to measure the surface potential difference between common inclusions from the nitinol matrix and the results indicate that the inclusions are more electrochemically noble than the nitinol matrix. Band energy theory is used to model the electrochemical interface between the passive films of nitinol and the solution under different applied potential conditions. A mechanism for the strong pitting corrosion resistance of nitinol in saline solution is proposed. PMID- 21648067 TI - Different patterns of bone fixation with hydroxyapatite and resorbable CaP coatings in the rabbit tibia at 6, 12, and 52 weeks. AB - Applying bioactive coatings on orthopedic implants can increase the fixation and long-term implant survival. In our study, we compared a resorbable electrochemically deposited calcium phosphate coating (Bonit(r)) to a thin (40 MUm) plasma-sprayed hydroxyapatite (HA) coating, applied on grit-blasted screw shaped Ti-6Al-4V implants in the cortical region of rabbit tibia, implanted for 6, 12, and 52 weeks. The removal torque results demonstrated stronger bone-to implant fixation for the HA than Bonit-coated screws at 6 and 12 weeks. After 52 weeks, the fixation was in favor of the Bonit-coated screws, but the difference was statistically insignificant. Coat flaking and delamination of the HA with multinucleated giant cell activity and bone resorption observed histologically seemed to preclude any significant increase in fixation comparing the HA implants at 6 versus 12 weeks and 12 versus 52 weeks. The Bonit-coated implants exhibited increasing fixation from 6 to 12 weeks and from 12 to 52 weeks, and the coat was resorbed within 6 weeks, with minimal activity of multinucleated giant cells or bone resorption. A different fixation pattern was observed for the two coatings with a sharper but time limited increase in fixation for the HA-coated screws, and a slower but more steadily increasing fixation pattern for the Bonit-coated screws. The side effects were more serious for the HA coating and limiting the expected increase in fixation with time. PMID- 21648068 TI - Effect of N-(p-coumaroyl)serotonin and N-feruloylserotonin, major anti atherogenic polyphenols in safflower seed, on vasodilation, proliferation and migration of vascular smooth muscle cells. AB - SCOPE: The objective of this study is to investigate a vascular effect of N-(p coumaroyl)serotonin (CS) and N-feruloylserotonin (FS), major antioxidative indolic polyphenols in safflower seeds with anti-atherogenic properties, with emphasis on effects on vascular smooth muscle cells (VSMCs). METHODS AND RESULTS: Both CS and FS (each 10 to 100 MUM) relaxed rat femoral arteries, which were pre contracted by 10(-5) M phenylephrine or 50 mM KCl, independently of their endothelium. Both CS and FS also concentration-dependently inhibited the increase of cytosolic free Ca(2+) concentration ([Ca(2+) ](i) ) that was induced by KCl or 5-hydroxytryptamine in cultured rat VSMCs. Next, we examined the effects of CS and FS on platelet-derived growth factor (PDGF)-BB-evoked proliferation and migration of the VSMCs. Both CS and FS inhibited PDGF-BB-evoked proliferation and migration of the VSMCs in a concentration-dependent manner. They also inhibited PDGF-BB-induced phosphorylation of PDGF receptor beta and ERK1/2, and Ca(2+) release from sarcoplasmic reticulum in the VSMCs in a concentration-dependent fashion. CONCLUSION: These results indicated a possible vascular effect of CS/FS to inhibit the activation of VSMCs by blocking the increase of [Ca(2+) ](i) and/or blocking PDGF signaling. These may explain a part of anti-atherogenic mechanism that underlies their ability to improve vascular distensibility and to inhibit aortic hyperplasia. PMID- 21648069 TI - Role of membrane dynamics processes and exogenous molecules in cellular resveratrol uptake: consequences in bioavailability and activities. AB - In the fields of nutrition prevention and therapy treatment, numerous studies have reported interesting properties of trans-resveratrol (RSV), a natural polyphenol against pathologies such as vascular diseases, cancers, viral infections and neurodegenerative processes. These beneficial effects are supported by more studies showing the pleiotropic actions of RSV. Nevertheless, a crucial question concerning these effects is how the polyphenol, when applied to an organism, gains access to its targets. In this review, we focus on the biochemical and biological parameters involved in RSV transport, particularly the role of the phospholipid bilayer in RSV uptake (passive diffusion, carrier mediated transport) and of exogenous molecules modulating RSV transport and effects. The dynamic processes of the plasma membrane reveal the importance of the role of lipid composition in the fluidity, the lipid rafts in RSV endocytosis and the ATP-binding cassette transporters in RSV efflux. Specific membrane receptors such as integrin alphavbeta3 contribute to RSV uptake and to activate signalling pathways involved in apoptosis. We discuss the role of intracellular receptors (i.e. aryl-hydrocarbon and estrogen receptors). In addition, circulating molecules (i.e. albumin, haemoglobin, fatty acids, lipoproteins) play a role as RSV carriers. Finally, we developed a hypothesis concerning the relation between RSV uptake and its biological activities. PMID- 21648070 TI - Extrusion cooking with glucose supplementation of fumonisin-contaminated corn grits protects against nephrotoxicity and disrupted sphingolipid metabolism in rats. AB - SCOPE: Fumonisin B1 (FB1) is a mycotoxin found in maize and maize-based foods. It causes animal diseases and is a suspected risk factor for cancer and birth defects in humans. Extrusion cooking reduces FB1 concentrations in maize however toxicity caused by unknown degradation or FB1-matrix reaction products might persist. METHODS AND RESULTS: To test the efficacy of extrusion to reduce FB1 toxicity, Fusarium verticillioides fermented corn (= maize) grits (Batch-1= 9.7 ppm FB1; Batch-2= 50 ppm FB1) were extruded without (Batch-1E; Batch-2E) or with 10% glucose supplementation (Batch-1EG; Batch-2EG). FB1 concentrations were reduced 64% (Batch-2E) to 94% (Batch-1EG) after cooking. When the uncooked and processed grits were fed (50% w/w in rodent chow) to rats for up to 8 weeks, FB1 intakes averaged 354, 103, and 25.1 cg/kg body weight/day for Batch-1, Batch-1E and Batch-1EG and 1804, 698, and 222 cg/kg body weight/day for the Batch-2, Batch 2E and Batch-2EG, respectively. Nephrotoxicity including apoptotic lesions and elevated sphingoid base concentrations decreased in a dose-dependent manner in groups fed Batch-1, Batch-1E, Batch-2, Batch-2E, or Batch-2EG and was absent in the Batch-1EG group. CONCLUSION: Extrusion cooking, especially with glucose supplementation, is potentially useful to reduce FB1 concentrations and toxicity of FB1-contaminated maize. PMID- 21648072 TI - Self-assembly of hydrophilic homopolymers: a matter of RAFT end groups. AB - Unusual self-assembly behavior is observed for a range of hydrophilic homopolymers. This self-assembly behavior is contrary to the expected behavior of such hydrophilic polymers and instead mimics more commonly reported amphiphilic block copolymers. It is proposed that the unique combination of hydrophobic end groups at both the alpha and omega chain end accounts for this unusual self assembly behavior. Complex internal polymer micelles are spontaneously formed when hydrophilic homopolymer polyelectrolytes and neutral polymers (with a weight fraction of the hydrophobic end groups <10 wt%) are directly dissolved in water. The homopolymers, poly[2-(diethylamino)ethyl methacrylate], poly(N isopropylacrylamide), and poly(ethoxyethylacrylate) are synthesized by reversible addition-fragmentation chain-transfer (RAFT) polymerization using S'-1-dodecyl (S')-(alpha,alpha'-dimethyl-alpha"-acetic acid) trithiocarbonate (DDMAT) and its derivatives as chain transfer agents (CTAs). A range of polyelectrolyte homopolymers with different terminal groups are designed and synthesized, which under acidic aqueous solution direct the self-assembly to form well-defined nanostructures. This assembly behavior was also observed for neutral polymers, and it was determined that the structure of the hydrophobic end groups (and thus choice of RAFT CTA) are very important in facilitating this unusual self-assembly behavior of hydrophilic homopolymers. It is proposed that the functionality of commonly used CTAs such as DDMAT, can affect the solution association of the resultant homopolymers and can in fact afford ABA' type polymers, which can undergo self-assembly to form higher-order nanostructures. PMID- 21648073 TI - Measurement of the intrinsic thermal conductivity of a multiwalled carbon nanotube and its contact thermal resistance with the substrate. AB - The intrinsic thermal conductivity of an individual carbon nanotube and its contact thermal resistance with the heat source/sink can be extracted simultaneously through multiple measurements with different lengths of the tube between the heat source and the heat sink. Experimental results on a 66-nm diameter multiwalled carbon nanotube show that above 100 K, contact thermal resistance can contribute up to 50% of the total measured thermal resistance; therefore, the intrinsic thermal conductivity of the nanotube can be significantly higher than the effective thermal conductivity derived from a single measurement without eliminating the contact thermal resistance. At 300 K, the contact thermal resistance between the tube and the substrate for a unit area is 2.2 * 10(-8) m(2) K W(-1) , which is on the lower end among several published data. Results also indicate that for nanotubes of relatively high thermal conductance, electron-beam-induced gold deposition at the tube-substrate contacts may not reduce the contact thermal resistance to a negligible level. These results provide insights into the long-lasting issue of the contact thermal resistance in nanotube/nanowire thermal conductity measurements and have important implications for further understanding thermal transport through carbon nanotubes and using carbon nanotube arrays as thermal interface materials. PMID- 21648071 TI - The inhibitory effects of 5-hydroxy-3,6,7,8,3',4'-hexamethoxyflavone on human colon cancer cells. AB - SCOPE: Previously, we reported that 5-hydroxy-3,6,7,8,3',4'-hexamethoxyflavone (5HHMF), a polymethoxyflavone found in citrus peels, potently inhibited the growth of multiple human colon cancer cells. Herein, we further investigated the anti-cancer mechanisms of 5HHMF in human colon cancer cells. METHODS AND RESULTS: Colony formation assay revealed that 5HHMF dose dependently inhibited colony formation of multiple colon cancer cells. Western blot analysis demonstrated 5HHMF decreased nuclear beta-catenin levels and increased the E-cadherin level in a dose-dependent manner. 5HHMF also modified plasma membrane-associated proteins, such as K-Ras, EGFR, and their downstream effectors, such as Akt. Moreover, treatments with 5HHMF inhibited nuclear translocation of NF-kappaB, which may contribute to its anti-cancer effects. Add-back study showed that the inhibitory effect of 5HHMF was not associated with the production of reactive oxygen species (ROS). In addition, 5HHMF treatment inhibited the capillary tube formation of human umbilical vein endothelial cells (HUVECs) on matrigel, suggesting a potential anti-tumor angiogenesis effect. CONCLUSION: Our results demonstrated that 5HHMF suppressed multiple oncogenic molecular events in colon cancer cells. PMID- 21648074 TI - Engineered multifunctional nanocarriers for cancer diagnosis and therapeutics. AB - This article reviews advances in the design and development of multifunctional carbon-based and/or magnetic nanoparticle systems (or simply 'nanocarriers') for early cancer diagnosis and spatially and temporally controlled therapy. The critical issues in cancer diagnosis and treatment are addressed based on novel nanotechnologies such as real-time in-vivo imaging, drug storage and release, and specific cancer-cell targeting. The implementation of nanocarriers into animal models and the subsequent effectiveness in treating tumors is also reviewed. Recommendations for future research are given. PMID- 21648075 TI - Acrylate-facilitated cellular uptake of gold nanoparticles. PMID- 21648076 TI - Image-guided prostate cancer therapy using aptamer-functionalized thermally cross linked superparamagnetic iron oxide nanoparticles. AB - CG-rich duplex containing prostate-specific membrane antigen (PSMA) aptamer conjugated thermally cross-linked superparamagnetic iron oxide nanoparticles (TCL SPIONs) is reported as prostate cancer-specific nanotheranostic agents. These agents are capable of prostate tumor detection in vivo by magnetic resonance imaging (MRI) and selective delivery of drugs to the tumor tissue, simultaneously. The prepared PSMA-functionalized TCL-SPION via a hybridization method (Apt-hybr-TCL-SPION) exhibited preferential binding towards target prostate-cancer cells (LNCaP, PSMA+) in both in vitro and in vivo when analyzed by T(2) -weighted MRI. After Dox molecules were loaded onto the Apt-hybr-TCL SPION through the intercalation of Dox to the CG-rich duplex containing PSMA aptamer as well as electrostatic interaction between the Dox-and-polymer coating layer of the nanoparticles, the resulting Dox@Apt-hybr-TCL-SPION showed selective drug-delivery efficacy in the LNCaP xenograft mouse model. These results suggest that Dox@Apt-hybr-TCL-SPION has potential for use as novel prostate cancer specific nanotheranostics. PMID- 21648077 TI - Liquid chromatography-mass spectrometric method for determination of the non imidazole H3-receptor antagonist UPR1056 in rat plasma. AB - The non-imidazole H3 receptor antagonist UPR1056 was dosed in plasma samples from rats individually administered with a single i.p. dose of 1.25 mg/kg by means of a newly validated HPLC-MS method. UPR1056 was extracted from rat plasma by protein precipitation with acetonitrile and was separated by linear gradient elution, employing water and methanol both additioned with 0.05% trifluoroacetic acid as mobile phases. UPR1056 was detected in MS using an electrospray ion source operating in positive ion mode. Acquisition was performed in single ion monitoring mode at m/z=349.3. The method was validated over the concentration range of 17.43-1743 ng/mL (50-5000 pmol/mL). Within- and between-run precision for the low, mid and high quality controls (QC) levels were 6.75% or less and accuracy ranged from 95.8 to 107.6%. The lower limit of quantification was 17.43 ng/mL. The analysis of the time course of UPR1056 concentrations over the 24-h period revealed a C(max) of 1147 ng/mL after 2 h from peripheral administration of the non-imidazole H(3)-receptor antagonist, with a prolonged elimination half life of over 9 h. PMID- 21648078 TI - Comparison of monolithic and 1.8-MUm RP-18 silica capillary columns using chromatographic data and mass spectrometric identification scores for proteins. AB - A goal in proteomics is the analysis of proteins by LC-MS. The proteins are enzymatically digested and the resulting peptides are chromatographically separated and introduced into a tandem MS. The obtained MS data are used for a search in sequence databases, providing identification scores for the proteins. A method to improve that score is to increase the chromatographic separation and peak capacity. In this study, the chromatographic conditions were optimized for a relatively large gradient time by varying the flow rate and gradient composition. The influence of the monolithic column length (15 and 64 cm) and particle diameter (1.8 MUm; 15-cm length) on the sample peak capacity, productivity and identification score was studied. For comparison of gradient systems, a scaling factor was introduced to normalize the properties/performance of columns for material, diameter and length. As model proteins/digests, a simple (myoglobin) and a larger (BSA) protein were used. The smallest peak width, highest identification scores (54 and 89% for BSA and myoglobin, respectively) and productivity (5.0 and 4.0, respectively) were obtained for the 15-cm particulate column. The study also demonstrates that a further increase in the chromatographic performance is beneficial for BSA but hardly increases the identification score for the relatively small myoglobin. PMID- 21648079 TI - Morphology-transport relationships for silica monoliths: from physical reconstruction to pore-scale simulations. AB - This work describes individual steps of an approach toward quantitative correlations between morphological and mass transport properties of capillary silica monoliths. The macropore space morphology of the central core region of the capillary monolith is visualized by a fast, nondestructive, and quantitative method using three-dimensional reconstruction from confocal laser scanning microscopy images. The reconstructed 60 MUm*60 MUm*12 MUm monolith domain consisted of 1.6*10(9) cubic voxels with 30 nm edge length. The received morphological data were chord length distributions for the bulk macropore space and skeleton of the monolith, which we characterized by k-gamma distributions. This analysis provides parameters that can be correlated with the mass transport properties obtained by macropore-scale simulations of flow and transport in the reconstructed monolith. These simulations were realized on a supercomputing platform and comprised the lattice-Boltzmann method for fluid flow and a random walk particle-tracking method for advective-diffusive mass transport. The characteristic length scales of eddy dispersion correlate with the statistical measures of the chord length distributions. Simulated plate height curves demonstrate that the bulk monolith is very homogeneous, and that the intraskeleton transport properties and a stochastic variation of macropore space characteristics can be neglected compared with the importance of reducing column radial heterogeneity in chromatographic practice. PMID- 21648080 TI - Simultaneous extraction of acidic and basic drugs from urine using mixed-mode monolithic silica spin column bonded with octadecyl and cation-exchange group. AB - A method coupling spin column extraction with gas chromatography-mass spectrometry was developed for the simultaneous extraction of acidic and basic drugs from urine. Benzodiazepines, local anaesthetics, antidepressants, and barbiturates were used as model drugs. Sample loading, washing, and elution of the target drugs were accomplished by centrifugation of the column. In this study, mixed-mode monolithic silica bonded with a C18 reversed-phase and a strong cation exchange phase was packed in a spin column. The pH of a urine sample (0.2 mL) was adjusted to 3 and the analytes adsorbed onto the column were eluted with 0.1 mL of MeOH containing 2% NH3; all the tested drugs were simultaneously extracted from urine. The recovery of the tested drugs was 65-123%. Up to a concentration of 2500 ng/mL of the target drugs in urine, a linear curve was observed (r(2)>0.996). The intra- and interday RSDs at three different concentrations in urine were 2.1-14.7%. For RSDs lower than 15%, the limits of detection were 1-25 ng/mL. The proposed method was successfully applied for clinical and forensic cases and the results thus obtained were in good agreement with those obtained by conventional methods. PMID- 21648081 TI - Monoliths with immobilized zirconium ions for selective enrichment of phosphopeptides. AB - To meet the demands of protein phosphorylation study, immobilized zirconium ion affinity chromatography (Zr(4+)-IMAC) monolith was prepared by combining UV initiated polymerization of monolithic support and subsequent photografting in both capillary columns and microchannels. Hydrophilic poly(2-hydroxyethyl methacrylate (HEMA)-co-ethylene dimethacrylate (EDMA)) monolithic support was prepared under UV irradiation at the wavelength of 365 nm with monomer HEMA, crosslinker EDMA and 2,2-dimethoxy-2-phenylacetophenone as photoinitiator in 1 decanol solution, which provides good biocompatibility and permeability for biomolecule analysis. To introduce chelating ligands, such as phosphate groups, on the pore surface of monolith for metal ion immobilization, photografting of ethylene glycol methacrylate phosphate with benzophenone as the photoinitiator was performed at 254 nm for 300 s. The grafting process and metal ion immobilization can be monitored by measuring the electroosmotic flow produced by the modified monolith, providing a quantitative evaluation of post-modification. This new method for the preparation of Zr(4+)-IMAC monolith simplifies the optimization of monolith preparation and avoids the time-consuming chemical modification process. Additionally, advantages include facile preparation in microdevices, easy regenerability and good reproducibility. After optimization, the microchip-based Zr(4+)-IMAC monolith was used for phosphopeptide analysis and showed good selectivity in phosphopeptide enrichment with matrix-assisted laser desorption ionization mass spectrometry detection. PMID- 21648082 TI - Optimization of lytic phage manufacturing in bioreactor using monolithic supports. AB - A process for manufacturing large quantities of lytic bacteriophages was developed. Determination of cultivation termination was found to be essential to achieve high phage quantity and purity. When optimal cultivation termination is missed, phage fraction was found to be highly contaminated with deoxyribonucleic acid released from Escherichia coli cells. Besides, an already established method for monitoring of phage cultivation based on optical density, where its peak indicates point when maximal phage titer is achieved, a new indirect chromatographic method using methacrylate monoliths is proposed for on-line estimation of phage titer. It is based on the measurement of released E. coli deoxyribonucleic acid and shows high correlation with phage titer obtained from plaque assay. Its main advantage is that the information is obtained within few minutes. In addition, the same method can also be used to determine purity of a final phage fraction. Two strategies to obtain highly pure phage fractions are proposed: an immediate purification of phage lysate using monolithic columns or an addition of EDTA before chromatographic purification. The developed protocol was shown to give phage purity above 90% and it is completed within one working day including cultivation and phage titer in the final formulation using developed chromatographic method. PMID- 21648083 TI - Preparation of macroporous cryostructurated gel monoliths, their characterization and main applications. AB - Cryostructuration platform renders it possible to form macroporous materials (known as cryogels) with a broad range of porosity, from structures with combination of meso- and macropores to structures with 100-MUm sized macropores. When these materials are formed in the shape of monoliths (monolithic cryogels), they present a unique monolithic stationary medium for specific applications. This review summarizes the recent research on the preparation and characterization of cryostructurated monolithic cryogels for (bio)separation and points to some future perspectives. PMID- 21648086 TI - Best practice in statistical reporting. PMID- 21648087 TI - Biomarker discovery from the top down: Protein biomarkers for efficient virus transmission by insects (Homoptera: Aphididae) discovered by coupling genetics and 2-D DIGE. AB - Yellow dwarf viruses cause the most economically important virus diseases of cereal crops worldwide and are vectored by aphids. The identification of vector proteins mediating virus transmission is critical to develop sustainable virus management practices and to understand viral strategies for circulative movement in all insect vectors. Previously, we applied 2-D DIGE to an aphid filial generation 2 population to identify proteins correlated with the transmission phenotype that were stably inherited and expressed in the absence of the virus. In the present study, we examined the expression of the DIGE candidates in previously unstudied, field-collected aphid populations. We hypothesized that the expression of proteins involved in virus transmission could be clinically validated in unrelated, virus transmission-competent, field-collected aphid populations. All putative biomarkers were expressed in the field-collected biotypes, and the expression of nine of these aligned with the virus transmission competent phenotype. The strong conservation of the expression of the biomarkers in multiple field-collected populations facilitates new and testable hypotheses concerning the genetics and biochemistry of virus transmission. Integration of these biomarkers into current aphid-scouting methodologies will enable rational strategies for vector control aimed at judicious use and development of precision pest control methods that reduce plant virus infection. PMID- 21648090 TI - The role of dimension in multivalent binding events: structure-activity relationship of dendritic polyglycerol sulfate binding to L-selectin in correlation with size and surface charge density. AB - L-, P-, and E-Selectin are cell adhesion molecules that play a crucial role in leukocyte recruitment from the blood stream to the afflicted tissue in an acute and chronic inflammatory setting. Since selectins mediate the initial contact of leukocytes to the vascular endothelium, they have evolved as a valuable therapeutic target in diseases related to inflammation by inhibition of the physiological selectin-ligand interactions. In a previous study, it was demonstrated that dPGS, a fully synthetic heparin analogue, works as an efficient inhibitor towards L- and P-selectin in vitro as well as in vivo. Herein, the focus is directed towards the effect of size and charge density of the polyanion. The efficiency of L-selectin inhibition via an SPR-based in vitro assay and a cell-based flow chamber assay is investigated with dPGS ranging from approximately 4 to 2000 kDa. SPR measurements show that the inhibitory potential of highly sulfated dPGS increases with size and charge density. Thereby, IC(50) values from the micromolar to the low picomolar range are determined. The same tendency could be observed in a cell-based flow chamber assay with three representative dPGS samples. This structure-affinity relationship of dPGS suggests that the strong inhibitory potential of dPGS is not only based on the strong electrostatic interaction with areas of cationic surface potential on L selectin but is also due to a steric shielding of the carbohydrate binding site by large, flexible dPGS particles. PMID- 21648091 TI - Adaptation of the "Dynamic Method" for measuring the specific respiration rate in oxygen transfer systems through diffusion membrane. AB - Monitoring the specific respiration rate (Q(O2)) is a valuable tool to evaluate cell growth and physiology. However, for low Q(O2) values the accuracy may depend on the measurement methodology, as it is the case in animal cell culture. The widely used "Dynamic Method" imposes serious difficulties concerning oxygen transfer cancellation, especially through membrane oxygenation. This paper presents an improved procedure to this method, through an automated control of the gas inlet composition that can minimize the residual oxygen transfer driving force during the Q(O2) measurement phase. The improved technique was applied to animal cell cultivation, particularly three recombinant S2 (Drosophila melanogaster) insect cell lines grown in a membrane aeration bioreactor. The average measurements of the proposed method reached 98% of stationary liquid phase balance method, taken as a reference, compared to 21% when the traditional method was used. Furthermore, this methodology does not require knowledge of the volumetric transfer coefficient k(L)a, which may vary during growth. PMID- 21648092 TI - A three-dimensional colocalization RNA interference screening platform to elucidate the alternative lengthening of telomeres pathway. AB - A high-content colocalization RNA interference screen based on automatic three color confocal fluorescence microscopy was developed to analyze the alternative lengthening of telomeres (ALT) pathway. Via this pathway telomerase-negative cancer cells can maintain their telomeres and with it their unlimited proliferative potential. A hallmark of ALT cells is the colocalization of promyelocytic leukemia (PML) nuclear bodies with telomeres to form ALT-associated PML nuclear bodies (APBs). In our screen, the presence of APBs was used as a marker to identify proteins required for the ALT mechanism. A cell-based assay and an automatic confocal image acquisition procedure were established. Using automatic image analysis based on 3D parametric intensity models to identify APBs, we conducted an unbiased and quantitative analysis of nine different candidate genes. A comparison with the literature and manual analysis of the gene knockdown demonstrates the reliability of our approach. It extends the available repertoire of high-content screening to studies of cellular colocalizations and allows the identification of candidate genes for the ALT mechanism that represent possible targets for cancer therapy. PMID- 21648093 TI - Minimal cells: relevance and interplay of physical and biochemical factors. AB - Research on the construction of minimal cell-like systems is continuously progressing. The aim is to assemble a synthetic or semi-synthetic cell by encapsulating the minimal set of different macromolecules into a lipid vesicle (liposome). Synthetic cells have their relevance as new biotechnological tool for use in synthetic biology and in research into the origin of life. In recent years, several technical advances have been reported and reviewed, but most deal with the biochemical and molecular biology of protein synthesis inside liposomes, whereas a discussion on the importance and the interplay of some physical factors has not been discussed. In this short review, we comment on physical aspects, such as compartment formation and solute entrapment, and on the nature of lipid membrane. Emphasis is given to their relevance for the technology of construction of synthetic cells, and for new aspects of vesicle population studies. PMID- 21648094 TI - Genetically engineered light sensors for control of bacterial gene expression. AB - Light of different wavelengths can serve as a transient, noninvasive means of regulating gene expression for biotechnological purposes. Implementation of advanced gene regulatory circuits will require orthogonal transcriptional systems that can be simultaneously controlled and that can produce several different control states. Fully genetically encoded light sensors take advantage of the favorable characteristics of light, do not need the supplementation of any chemical inducers or co-factors, and have been demonstrated to control gene expression in Escherichia coli. Herein, we review engineered light-sensor systems with potential for in vivo regulation of gene expression in bacteria, and highlight different means of extending the range of available light input and transcriptional output signals. Furthermore, we discuss advances in multiplexing different light sensors for achieving multichromatic control of gene expression and indicate developments that could facilitate the construction of efficient systems for light-regulated, multistate control of gene expression. PMID- 21648095 TI - Efficient one-step synthesis of bis-spiroketals from diynediols by pi-Lewis acid catalyzed hydroalkoxylation/hydration. PMID- 21648096 TI - Cobalt monolayer islands on Ag(111) for ORR catalysis. AB - The design of a catalyst for one of the most important electrocatalytic reactions, the oxygen reduction reaction (ORR), was done following the most recent guidelines of theoretical studies on this topic. Aim of this work was to achieve a synergic effect of two different metals acting on different steps of the ORR. The catalytic activity of Ag, already known and characterized, was enhanced by the presence of a monolayer of cobalt subdivided into nanosized islands. To obtain such a controlled nanostructure, a novel method utilizing self assembled monolayers (SAMs) as templates was employed. In a recent study, we were able to perform a confined electrodeposition of cobalt onto Ag(111) in a template formed by selectively desorbing a short-chain thiol (3-mercaptopropionic acid, MPA) from binary SAMs using 1-dodecanthiols (DDT). This method allows for an excellent control of the morphology of the deposit by varying the molar ratio of the two thiols. Because cobalt does not deposit on silver at an underpotential, the alternative approach of surface limited redox replacement (SLRR) was used. This method, recently developed by Adzic et al., consists of the use of a monolayer of a third metal, which can be deposited at an underpotential, as a template for the spontaneous deposition of a more noble metal. Herein, we choose zinc as template for the deposition of cobalt. Ag(111) crystals were covered by monolayer islands consisting of cobalt, with the surface atomic ratios ranging from 12 to 39% for cobalt. The catalytic activity of such samples towards ORR was evaluated and the best improvement in activity was found to be that of the sample with a cobalt percentage of approximately 30% with respect to the bare silver, which is in good agreement with theoretical hypotheses. PMID- 21648101 TI - Effects of IMAC specific peptide tags on the stability of recombinant green fluorescent protein. AB - Immobilized metal ion affinity chromatography (IMAC) using peptide affinity tags has become a popular tool for protein purification. An important feature dictating the use of a specific affinity tag is whether its structure influences the properties of the target protein to which it is attached. In this work we have studied the influence on protein stability of two novel peptide affinity tags, namely NT1A and HIT2, and compared their effect to the commonly used hexa histidine tag, all attached to the C-terminus of a enhanced green fluorescent protein (eGFP). A comparison of the influence of C- or N-terminal orientation of the tags was also carried out by studying the NT1A tag attached at either terminus of the eGFP. Protein stability was studied utilising guanidine hydrochloride equilibrium unfolding procedures and CD and fluorescence spectroscopy. The novel peptide affinity tags, NT1A and HIT2, and the His6 tag were found to not affect the stability of eGFP. Although these results are protein specific, they highlight, nevertheless, the need to employ suitable characterisation tools if the impact of a specific peptide tag on the folded status or stability of a recombinant tagged protein, purified by immobilized metal ion affinity chromatographic methods, are to be rigorously evaluated and the appropriate choice of peptide tag made. PMID- 21648102 TI - Effects of temperature and pH on growth and antioxidant content of the microalga Scenedesmus obliquus. AB - Reactive forms of oxygen can damage DNA (among other molecules), thus triggering, e.g., atherogenesis and carcinogenesis. However, such dietary antioxidants as lutein and beta-carotene can effectively inactivate them; these compounds were found to high levels in a novel strain (M2-1) of the microalga Scenedesmus obliquus. The independent and combined effects of pH and temperature on its rates of growth and production of antioxidants were experimentally assessed, via a full factorial experimental design; the effects of each parameter independently, and of their interactions were accordingly quantified by ANOVA. Our results indicated that temperature plays a more important role on the maximum specific growth rate than pH; in terms of antioxidant content, pH and, to a lesser extent, temperature also have relevant effects. Consequently, the highest rate of biomass specific growth (0.294 +/- 0.013 day(-1)) and biomass productivity (0.837 +/- 0.054 mg L( 1) day(-1)) were associated with relatively low pH (6) and relatively high temperature (30 degrees C). Conversely, the antioxidant production rate increased with pH; hence, the highest productivity (0.638 mg L(-1) day(-1)) was attained at pH 8 and 30 degrees C. At the best operating conditions for antioxidant content, the levels of lutein and beta-carotene were 203.57 +/- 1.41 and 18.20 +/- 0.33 mg mL(-1), respectively; the maximum production of either one occurred at the early exponential phase. PMID- 21648103 TI - ["Unclassified" renal cell carcinomas]. PMID- 21648104 TI - [Online physician search of the statutory health insurance. How how can benefit from this site]. PMID- 21648105 TI - [Controversial "irritable bowel" topic. No incidental diagnosis, but a diagnosis by exclusion]. PMID- 21648106 TI - [Epidemiology and aetiology of female urinary incontinence]. PMID- 21648107 TI - [Better medical care in nursing homes is possible, but remuneration should be appropriate (interview by Dr. med. Horst Gross)]. PMID- 21648108 TI - [The impact of lifestyle factors on male reproductive health]. PMID- 21648109 TI - ["Beautiful new world of health care": apps also conquer medicine]. PMID- 21648110 TI - [Neurologic "daily bread" for the family physician. With hammer, needle and fingertip sensitivity toward diagnosis]. PMID- 21648111 TI - [Calcifying tendinitis of the shoulder]. PMID- 21648112 TI - [Use in general practice also appealing. The pc tablet: flexible solutions for the physician]. PMID- 21648113 TI - [Therapy of knee osteoarthritis. From elevated shoe sole to endoprosthesis]. PMID- 21648114 TI - Hot, blue, suspicious: comment on "Processing sentinel nodes in breast cancer". PMID- 21648115 TI - Clarity, confusion, or conundrum: comment on "Trends in diverticulitis management in the United States from 2002 to 2007". PMID- 21648116 TI - Sex differences for traumatic brain injury outcomes: comment on "Protection from traumatic brain injury in hormonally active women vs men of a similar age". PMID- 21648117 TI - Serum vitamin D concentrations and associated severity of acute lower respiratory tract infections in Japanese hospitalized children. AB - BACKGROUND: Vitamin D is an immunomodulatory molecule related to innate immunity that may contribute to the increased occurrence of acute lower respiratory infection (ALRI) in children, one of the most common reasons for hospitalization and intensive care unit admission. In the present study, the association between vitamin D deficiency and the severity of respiratory infection was evaluated by determining serum concentrations of 25-hydroxyvitamin D (25(OH)D) in a group of hospitalized children with ALRI. METHODS: Of the 28 children admitted to Nihon University Nerima-Hikarigaoka Hospital with ALRI over the period November 2008 May 2009, 26 were diagnosed as having bronchiolitis and two were found to have pneumonia. A competitive protein binding radioimmunoassay was used to determine serum 25(OH)D concentrations. RESULTS: Mean 25(OH)D concentrations in breast-fed children with ALRI (n = 7) were significantly lower than those in children with ALRI who were bottle fed/weaned (n = 6) or on a regular diet (n = 15; 14.6 +/- 9.7, 28.9 +/- 6.9 and 24.6 +/- 8.8 ng/mL, respectively). There was a significant correlation between vitamin D deficiency (<15 ng/mL) and the need for supplementary oxygen and ventilator management. CONCLUSION: Significantly more children with ALRI who needed supplementary oxygen and ventilator management were vitamin D deficient. These findings suggest that the immunomodulatory properties of vitamin D may influence the severity of ALRI. PMID- 21648118 TI - Japanese Guidelines for the Management of Respiratory Infectious Diseases in Children 2007 with focus on pneumonia. AB - Members of the Japanese Society of Pediatric Pulmonology and the Japanese Society for Pediatric Infectious Diseases developed the Guidelines for the Management of Respiratory Infectious Diseases in Children with the objective of facilitating the appropriate diagnosis and treatment of childhood respiratory infections. To date, a first edition (2004) and a revised edition (2007) have been issued. Many problems complicate the diagnosis of the pathogens responsible for bronchopulmonary infections in children. The Guidelines were the first pediatric guidelines in the world to recommend treatment with antimicrobials suited to causative pathogens as identified from cultures of sputum and other clinical specimens collected from infection sites and satisfying assessment criteria. The major causative microorganisms for pneumonia in infants and children were revealed to be Streptococcus pneumoniae, Haemophilus influenzae and Mycoplasma pneumoniae. This manuscript describes the Guidelines for the Management of Respiratory Infectious Diseases in Children in Japan 2007, with a focus on pneumonia. PMID- 21648119 TI - Response to letters regarding article, "Predictors of outcomes in medically treated patients with acute coronary syndromes after angiographic triage: an Acute Catheterization and Urgent Intervention Triage Strategy (ACUITY) substudy". PMID- 21648120 TI - China's organ transplant system in transition. PMID- 21648121 TI - Ethiopia gears up for more major health reforms. PMID- 21648122 TI - The quiet art revisited. PMID- 21648123 TI - The prevalence and clinical characteristics of punding in Parkinson's disease. AB - BACKGROUND: Punding (the display of stereotyped, repetitive behaviors) is a relatively recently discovered feature of Parkinson's disease (PD). Little is known about the prevalence and clinical characteristics of punding in PD. METHODS: In this review, four large scientific databases were comprehensively searched for literature in relation to punding prevalence and clinical correlates in the context of PD. RESULTS: Prevalence was found to vary greatly (between 0.34 to 14%), although there were large disparities in study populations, assessment methods, and criteria. We observed an association between punding, dopaminergic medications, and impulse control disorder. Other characteristics, which may be more common among punders, include a higher severity of dyskinesia, younger age of disease onset, longer disease duration, and male gender. DISCUSSION: More research in large clinical datasets is required in many areas before conclusions are drawn. The pathophysiology behind the punding phenomenon is also poorly understood at present, rendering it difficult to develop targeted therapy. The current mainstay of treatment is the reduction in the dose of dopaminergic medications, the evidence for other suggested therapies being purely empirical. PMID- 21648124 TI - Progression of premanifest and early Huntington's disease detectable after 1 year does TRACK-HD open the door to disease-modifying trials in HD and beyond? PMID- 21648125 TI - International consortium identifies new genetic risk factors for Parkinson's disease. PMID- 21648126 TI - Community acquired pneumonia in Port Harcourt Rivers State of Nigeria. AB - BACKGROUND: Community acquired pneumonia (CAP) is a major cause of death world wide. Knowledge of the likely pathogens and their sensitivity/resistance pattern can help in the choice of antibiotic therapy and improve outcome. OBJECTIVE: To identify the seasonal variation; age and sex distribution; bacteriology; antimicrobial sensitivity pattern of isolates; haematological data; radiology and clinicaloutcome of community-acquired pneumonia (CAP) in adult patients admitted to hospital in Port Harcourt, Nigeria. DESIGN: Prospective study. SETTING: University of Port Harcourt Teaching Hospital, and Braithwaite Memorial Hospital, Port Harcourt, Rivers State. PATIENTS: Adults admitted to the hospitals with CAP between 1 May 2002, and 30 April 2003. INTERVENTIONS: A diagnostic strategy using regular collection of sputum samples for gram stain, bacteriological culture and chest radiography were done. Blood cultures were done in severe cases. Antibiotic sensitivity testing was done on the positive cultures. RESULTS: During a 12 month period, 54 patients aged 16 to 82 years (mean 38.1 years) were evaluated. A total of 944 medical admissions were seen during the same period. This gives a prevalence rate of 5.7%. Twenty six classes of bacteria were isolated from the sputum of 23 patients (yield, 42.6%). Streptococcus pneumoniae, the most common pathogen, was isolated in nine cases (34.6%), followed by Klebsiella pneumoniae which was present in eight (30.8%). Other isolates included, Escherichia coli, four cases (15.4%), Pseudomonas aeruginosa, three cases (11.5%), and Staphylococcus aureus, two cases (7.7%). The commonest radiological pattern was lobar consolidation (49.9%) with no Distinct pattern associated with any conventional bacterial pathogen. The isolates showed good sensitivity to the newer and more expensive antibiotics (quinolines and cephalosporines) with marked resistance to the older and cheaper ones. However, clinical responses to benzyl penicillin and gentamycin were found to be good. Out of 54 patients evaluated, four (7.4%) died. CONCLUSION: Streptococcus pneumoniae and Klebsiella were common aetiological organisms of CAP in Port Harcourt. Treatment of CAP with benzyl penicillin remains an appropriate first line choice in this environment while the more expensive quinolines and cephalosporines can be used as backups. PMID- 21648127 TI - [Multiple suppurative arthritis, intervertebral discitis, musculus iliopsoas abscess, bacteremia caused by G group hemolytic streptococcus]. PMID- 21648128 TI - Last call: HIV patients endangered by smoking, drinking alcohol. HIV + age + risk behaviors = heart, lung disease. PMID- 21648129 TI - Special report: HIV smoking and drinking. A striking majority in HIV cohort smoke. Five A plan: ask, advise, assess, assist, arrange. PMID- 21648130 TI - Special report: HIV smoking and drinking. Pregnant pause: HIV and alcohol don't mix. Alcohol a transport molecule for HIV. PMID- 21648132 TI - Female condoms: Empowered prevention. Market rapidly expands, use triples. PMID- 21648131 TI - CDC issues new guidelines for community centers. Routine HIV testing, counseling update. PMID- 21648133 TI - Nearing 30th anniversary of HIV/AIDS in the U.S. Reflection, sharing encouraged. PMID- 21648134 TI - FDA notifications. Abacavir sulfate tablets tentatively approved. PMID- 21648135 TI - [Generalized exanthema in a 14-year-old boy: is it contagious?]. PMID- 21648136 TI - [Current strategies for the treatment of allergies. Well protected against pollen?]. PMID- 21648137 TI - [New medicinal treatment options for inhalation allergies of the upper airways]. PMID- 21648138 TI - [Cluster immunotherapy in allergic rhinoconjunctivitis]. PMID- 21648139 TI - [Emergency checklist: Hypothermia]. PMID- 21648140 TI - [Pseudoxanthoma elasticum--case report]. PMID- 21648141 TI - [Mature cystic teratoma--two decades of pain]. PMID- 21648142 TI - [Complementary medicine and asthma patients]. PMID- 21648143 TI - [Therapy with pentaerithrityltetranitrate (PETN). Established effectiveness, once again on trial]. PMID- 21648144 TI - [Contemporary possibilities of artificial vision in blind patients using artificial neuro-prosthesis--review]. AB - The authors present the knowledge summarization of vision substitute using artificial retinal neuroprostheses. The overview was compiled from available literature found in the Medline and Embase databases. The text focuses on global summarizing of the whole issue since the beginning of the development to the state up to date. In individual types of neuroprostheses, the advantages, disadvantages, and possible obstacles to their use in clinical practice are discussed. Furthermore, the brain plasticity and functional changes of the brain in blinds are considered. The aim of the review is not to present all information in detail, but complex overview with relevant literature sources. PMID- 21648145 TI - [Long term efficiency of a deep sclerectomy with T-flux implant]. AB - PURPOSE: To evaluate a long term efficiency of a deep sclerectomy with T-Flux implant on intraocular pressure METHODS: Retrospective noncomparative analysis of glaucomatic patients from our ambulance who underwent deep sclerectomy combined with T-flux implantation. The main postoperative outcome measures were intraocular pressure (IOP), number of treatments and Nd:YAG goniopunctures. RESULTS: Mean preoperative IOP was 26.7 (+/- 0.8) mmHg, mean postoperative IOP was 17.4 (+/- 0.8) at month 6, 16.5 (+/- 0.7) at month 12, 16.9 (+/- 0.9) at month 24, 17.2 (+/- 1.0) at month 36, 18.6 (+/- 1,1) at month 48, 17.6 (+/- 1.1) at month 60 and 18.3 (+/- 1.8) at month 72. Mean preoperative number of medications per patient was reduced from 2.5 (+/- 0.13) to 0.73 (+/- 0.13) at month 6, 1.02 (+/- 0,13) at month 12, 1.1 (+/- 0.15) at month 24, 1.0 (+/- 0.18) at month 36, 1.22 (+/- 0.18) at month 48, 1.68 (+/- 0,2) at month 60, 1.8 (+/- 0.3) at month 72. Four patients (10%) underwent early goniopuncture and six patients (15%) late goniopuncture. CONCLUSION: Deep sclerectomy combined with T Flux implant appears to be a relatively safe antiglaucomatic surgery with a minimum number of complications, high predictability of the result and a very good long term effectivity. PMID- 21648146 TI - [Cardiovascular diseases occurrence in patients with pseudoexfoliative syndrome and pseudoexfoliative glaucoma]. AB - A retrospective evaluation of cardiovascular disease (CV disease) in patients with pseudoexfoliative syndrome (PEX syndrome) and pseudoexfoliative glaucoma (PEX glaucoma) and their relationship to morphological and functional eye parameters. The main objective of this comparison was to identify parameters, that are among the groups with severe and mild CV disease statistically significantly different. METHODS AND RESULTS: Retrospective analysis of patients with PEX syndrome and PEX glaucoma. Evaluated were 46 eyes of 46 patients equally divided into two groups. In the group with PEX glaucoma and in the group with PEX syndrome was evaluated as an equivalent number of 23 patients. Monitored and statistically evaluated parameters were age, sex, the value of best corrected visual acuity, intraocular pressure, central corneal thickness, the cup/disc ratio, perimetry parameters MD (Mean deviation) and PSD (Pattern standard deviation). Age, sex of patients and the average values of monitored parameters are displayed on descriptive evaluative chart. Using the Mann-Whitney test were compared the observed signs among patients with severe and mild CV disease in a group of PEX syndrome and PEX glaucoma. CV disease was classified as mild (typically middle and medically well-controlled hypertension or heart disease with no associated events such as myocardial infarction, stroke, heart failure) or severe (CV disease-related event such as myocardial infarction, cerebrovascular stroke, heart failure or venous occlusion). CONCLUSION: We did not find a statistically significant difference in the incidence of mild and severe CV diseases among groups of patients with PEX syndrome and PEX glaucoma. Also, we did not find statistical difference in the monitored parameters of visual acuity, intraocular pressure, central corneal thickness, cup/disc ratio and perimetric parameters MD and PSD between the groups with mild and severe CV problems. The exception was the cup/disc ratio parameter in the group of patients with PEX glaucoma, where we found a lower value of cup/disc ratio in patients with severe CV diseases than patients with mild CV diseases. PMID- 21648147 TI - [Electronic system for correct head position control after some vitreoretinal surgeries]. AB - Team of authors consisting of vitreoretinal surgeons and biomedical engineers developed and tested an electronic system helping the patients after some vitreoretinal surgeries to keep the recommended head position. The authors describe the principle of this system and its use in clinical practice. PMID- 21648148 TI - [Suggested low vision care for visually impaired children in Slovakia]. AB - INTRODUCTION: There is currently no system of registration for visually impaired children in Slovakia and the current prevalence of visual impairment (VI), low vision and blindness is unknown for this population. We propose a template for a process of registration of visually impaired children in Slovakia as well as a system for the Low Vision Health Core for this population. MATERIAL AND METHODS: Based on a literature search, we report our data of the estimated prevalence of VI in children in Slovakia and the number of registered students with VI. We have created a registration form and suggested a template of registration for VI children as well as the Health Care System for this population. RESULTS: In industrialized countries, the prevalence of VI, including blindness is 10 22/10,000 in children aged less than 16 years. Extrapolating these figures to the Slovak population, we estimate that there will be between 1500 to 3200 VI children under the age of 19 years. Only 752 students with VI of this age were recorded in Slovakia in 2009/2010. We suggest that three Low Vision Centres for VI children should be adequate to cater for the VI population, each of which should provide all levels of care and that ophthalmologists should register patients with VI by filling the proposed registration form. CONCLUSION: The number of VI children in Slovakia appears to be very low. The only way of accurately assessing the prevalence is to introduce a VI registration system into the country, to be carried out by the ophthalmologists. We suggest that the Low vision service provided by the health authority needs to be divided into primary, secondary and tertiary care (including visual rehabilitation by low vision aids). Only if the Health Insurance will adequately remunerate the Ophthalmologists for the individual procedures will they be motivated enough to provide this level of health care to VI patients. PMID- 21648149 TI - [Cataract after lightning injury--case report]. AB - Lightning injuries are dangerous and often deadly trauma. We present a case of 41 year old woman who suffered an indirect lightning injury, dropping into unconsciousness followed by amnesia. Initial irritation of her left eyeball was followed by subtle changes in retinal pigment epithelium without any drop in visual acuity for a period of three months. Subsequently there began a slow deterioration of visual acuity and progression of cataract causing BCVA to drop to 5/50. Patient underwent uncomplicated cataract phacoemulsification with IOL implantation which resulted in restoring BCVA to 5/5 postoperatively. Such cases are scarce in literature. PMID- 21648150 TI - [Value of McKenzie examination in clinical diagnosis of back pain caused by infection or metastasis]. AB - INTRODUCTION: Back pain caused by metastases and infections are rare and are called "red flags". The aim of this study conducted for physiotherapy needs was to answer questions asked during taking a subjective examination among patients with serious spine pathology. MATERIAL AND METHODS: Examinations conducted according to McKenzie method were compared. 17 patients were evaluated. In I group were 7 persons with metastasis, in II group 10 patients with infection of the spine. RESULTS: Results were statistically evaluated by Chi-square test. Exact Fisher test for small groups was used. The most characteristic for patients with metastasis were: constant pain, increasing of pain during movement, decreasing during lying, and poor health and weight loosing. For infection typical signs was fever accompanying pain. CONCLUSIONS: (1) Clinical examination plays key role in diagnostic. (2) Own surveys confirmed that McKenzie subjective examination enables physiotherapists to avoid traps in diagnosis of red flags. (3) Constant pain, weight loosening are characteristic for metastasis, and for infection constant pain and fever. PMID- 21648151 TI - [The value of femoral anteversion angle measured clinically and on radiographs]. AB - The purpose of the study was to analyze the value of clinical and radiological measurements of femoral anteversion. There were 49 patients (39 girls, 10 boys) with the mean age of 9.1 years (range from 4 to 15 years) at last follow-up. Among 98 analyzed hips 59 had IIb hip dysplasia according to Graf at childhood. Clinically femoral anteversion angle was measured using the trochanteric prominence angle test. On radiographs femoral anteversion angle was measured according to Strzyzewski method. Femoral anteversion wasn't significantly increased in joints type IIb. The correlation of clinical and radiographic determination of femoral anteversion was good (r=71) and it is recommended for routine examination. Hip rotation in flexion (r=59) better correlated with radiographic anteversion than in extension (r=46) and their combination increased correlation (r=62). Clinical examination in hips flexed 90 degrees allows for better assessment of femoral anteversion than examination done in hip extension. PMID- 21648152 TI - [Usefulness of HA+beta-TCP in bone defects repair during revision hip and knee arthroplasty]. AB - BACKGROUND: Bone deficiency can present a surgical challenge during revision total hip and knee arthroplasty. The amount of bone grafts available for surgical purposes is insufficient. Synthetic bone substitutes can eliminate the risk of infection transmission. The purpose of the present study was to evaluate the clinical and radiographic outcomes of revision hip and knee arthroplasty with use of an impaction bone-grafting with HA+beta-TCP in reconstruction of bone defects. MATERIALS AND METHODS: 20 total hip revision (THR) and 10 total knee revision (TKR) were performed in 30 patients where impaction bone-grafting technique with HA+beta-TCP was used. Clinical, radiographic and CT results were assessed. Mean follow-up was 21 (11-48) months for THR and 22 (10-46) months. RESULTS: Loosening was seen in 2 cups. In one case the reoperation was performed. In the second due to massive bone loss the prosthesis was removed. No another acetabular and stem components required revision surgery. There were no knee implant migration or loosening observed. The mean total HSS score was 45.3 preoperatively and 71.5 postoperatively. The mean CRS score was 35.7 preoperatively and 73.4 postoperatively. CONCLUSIONS: The use of HA+beta-TCP with bone grafts impaction is a good method of bone defect reconstruction and can provide good short-term clinical results in revision hip and knee arthroplasty. PMID- 21648153 TI - [Use of bulky, femoral head allografts in revision hip arthroplasty for acetabular component aseptic loosening]. AB - The aim of the study was to analyze results in patients treated with frozen, bulky femoral head allografts without reinforcement ring for significant bone lost after acetabular component aseptic loosening. Retrospective analysis was done on 19 patients in the average age of 58 years. There were 13 women in this group. Acetabular bone lost was classified as IIIa or IIIb according to Paprosky. For revision 10 cemented and 9 uncemented cups were used. Mean coverage of acetabular component by graft on antero-posterior radiographs was 52% (from 30% to 100%). The mean follow-up was 4.2 years. Seven of our patients required another revision for aseptic loosening of the cup. The remaining 12 patients had satisfactory clinical and radiological result. The function improved from 35 points before operation to 76 at last follow-up according to Harris grading system (minimum improvement was 20 points). Two of the patients had radiographic signs of osteolysis around implant, without symptoms of loosening. Coverage of acetabular cup by the graft was 65% in cases of loosening and 42% in those patient without loosening (p <0.01). There was no statistical relationship between age and frequency of loosening (p > 0.05). Bulky, femoral head allografts are passive scaffold and may lose the mechanical strength. In cases of big bone defects of supero-lateral part of acetabulum use of metal reinforcement rings should be considered. PMID- 21648154 TI - [Total hip replacement for acetabular protrusion in patients with osteoarthritis]. AB - BACKGROUND: Total hip replacement (THR) is at present an accepted treatment in patients with severe deformity of the hip and acetabular protrusion. The aim of this paper is to evaluate the results of THR in patients suffering from acetabular protrusion, operated from 1991 to 2006 in the Orthopaedic and Traumatologic Department of Poznan University of Medical Sciences. MATERIAL: Material included 36 patients, 34 females and 2 males, on whom 51 THR were performed, lateral approach was used in all cases. At the time of operation, the age of patients ranged from 41 to 78 years (mean 63). Follow-up ranged from 4 to 19 years (mean 9.9 years). The operative treatment was a multistage process (during one operation only one joint was replaced). Cemented total hip arthroplasty was used during 28 of the THR, 22 of them were cementless and 1 as a hybrid. METHOD: The patients were clinically and radiologically evaluated preoperatively, postoperatively, and at final examination. The clinical state was evaluated with Harris hip score and WOMAC scale. We based our radiological examination on Hip Society system. RESULTS: The average preoperative Harris score for the group of patients was 29, WOMAC score 77. After an average of 9 years follow-up all hips were considered excellent, with average Harris score of 90, WOMAC Score of 6. All patients had increased function and decreased pain. The radiograms of all patients revealed that the acetabular and femoral components were correctly positioned with no radiographic evidence of loosening in the last examination. The inclination angle of the acetabular component was 30-55 degrees (mean: 40 degrees) and the acetabular opening angle was 2-15 (mean: 4 degrees). The stem was valgus-oriented in 8 hips, varus-oriented in 13 hips and neutral oriented in 30 hips. No ectopic ossification concentrations were found. CONCLUSION: Clinical and radiological evaluation of our material showed that total hip replacement in the treatment of severe deformity of the hip caused by osteoarthritis with acetabular protrusion allows regaining good lower limb function, which helps the patients staying less dependant on the surrounding environment. The results of THR are good regardless of the type of prosthesis and the type of fixation. Ectopic ossification is not the clinical problem during THR in patients who suffer from acetabular protrusion. We found a significant acetabular remodeling with the decreasing of the protrusion after THR. PMID- 21648155 TI - [Arthroscopic fracture fixation of intercondylar eminence in children using instrumentarium for the reconstruction of anterior cruciate ligament]. AB - In our article we introduce a proposal of intercondylar tibial eminence fracture in children management. When dealing with II and III type fracture according to Mes and McKeever classification, we would like to suggest artroscopic fracture fixation with the help of a tension band wiring technique using single bundle reconstruction set of anterior cruciate ligament.The method mentioned above was presented on two cases managed in our ward. Stable fixation of this type fracture allows for quick mobilization and physiotherapy of a patient. It appears to be the key element to full recovery in articular surface fracture. PMID- 21648156 TI - [Survivorship of the cemented hip prosthesis]. AB - OBJECTIVES: To asses a survival-time of the cemented hip prostheses and to find factors causing its loosening. MATERIALS AND METHODS: 211 patients (282 hip prostheses) were analyzed, of the 1693 patients (2096 hip prostheses) treated between 1970-2000. The follow-up ranged from 7 to 34 years (mean: 12.2 ++/- 5.4 years). The age of the patients at the operation procedure ranged from 25 to 74 years (mean 52 +/- 12 years). Only aseptic prosthesis loosening were considered and Kaplan-Meyer method was used for the prosthesis survivorship assessment. RESULTS: Of the 282 prostheses, implant exchange was performed in 39 cases, where in 15 cases only cup was revised, in 5 cases the stem exclusively, and in 19 patients the whole implant was exchanged. Only 1.1% of implants were exchanged during first 5 years after surgery but from the 6th year the percentage of loosening increased several percents each year. Respectively 88.6, 70 and 65% survivorship of prosthesis was found after 10, 15, 20 years of follow-up. In the group of bilateral prosthesis implantation, the first implanted prosthesis was more susceptible for loosening. Seven cases of aseptic loosening (5.4%) of the 129 prostheses implanted between 1974 and 1995, occurred within 7 years after initial surgery where only one prosthesis (0.7%) was revised of the 152 implanted between 1996-2000 in the same time of follow-up. The age, growth, weight, BMI and sex did not influence the risk of implant loosening. CONCLUSION: The mean survivorship of the hip prosthesis is 65% in 20- years follow up. When a proper initial fixation of the cemented hip prosthesis is performed, the risk of implant loosening increases gradually especially from the 6th year after surgery. The modern implants are less susceptible for loosening. PMID- 21648157 TI - [Total knee replacement in joints with severe varus and bone deficiency]. AB - INTRODUCTION: Total knee replacement in knees with severe valgus and defects of the medial tibial condyle are at higher risk complications than in knees with a correct axial allignement. MATERIAL AND METHODS: The study group included 10 patients (11 knees) with with severe valgus and defects of the medial tibial condyle in which TKR was performed. In 8 cases bone defects were filled with bone autograft in 3 cases defects were filled with metal augments. The mean follow-up was 37 months (26 months to 4 years). Clinical results were measured with the knee society score, the level of activity was evaluated with the UCLA score. Radiographic evaluation was based on radiograms taken preoperatively, postoperatively and at the follow-up examination. RESULTS: In All cases an improvement in clinical results was noted, the mean KSS result rised from 25.3 preoperatively to 87 postoperatively and the level of activity rised from 3.4 to 5.4 postoperatively. No signs of loosening were found in radiographic evaluation. CONCLUSIONS: TKR in joint with severe varus and medial tibial condyle defects creates a need for the reconstruction of defects. In lesser defects reconstruction with bone autografts is sufficient, in severe cases filling the defect with metal augments is needed. When indicitions are followed both methods provide good clinical results. PMID- 21648158 TI - [The results of the treatment of thumb carpometacarpal arthritis by trapezium resection and modified thumb metacarpal suspension arthroplasty]. AB - Arthritis of the thumb carpometacarpal joint is a fairly common disorder of unclear etiology, affecting mostly women in the postmenopausal age. The results of the operative treatment of 21 patients, 20 women and 1 men in a mean age of 59 years with the disorder classified in 3rd and 4th grade in Eaton-Littler scale are presented. Operation consisted in resection of the trapezium followed by thumb metacarpal suspension arthroplasty with flexor carpi radialis tendon, using a slip of the palmaris longus tendon. At a mean follow-up of 14 months, patients achieved significant reduction of the pain at the movement of the thumb (VAS from a mean of 8.1 to 2.9), improvement in thumb's opposition (Kapandji test from 7.6 to 9.5), power of the hand increased form 42 to the 68% of the other side and function of the hand in DASH score improved form 58 to 12 points. Pinch strength did not improved after surgery. The results show effectiveness of the modified technique in the treatment of thumb carpometacarpal arthritis. PMID- 21648159 TI - [The usefulness of ultrasonography in the diagnosis of carpal tunnel syndrome--a review]. AB - Ultrasonography has had relatively long history (approximately 20 years) in the diagnosing carpal tunnel syndrome, but as late as in last decade has gained greater popularity and has been applied in the clinic. Numerous studies revealed that the most accurate parameter indicating the compression of the median nerve in the carpal tunnel is the cross sectional area of the nerve at the inlet level. However, contrary to the nerve conduction studies, sonographic measurements are characterised by wide range of normal, physiological (a mean of 4.8 to 9.7 mm2), pathological, indicating compression of the nerve values (a mean of 10.7 to 16.8 mm2) and cut-off coefficients between normal state and pathology (a mean of 6.5 to 14 mm2). Sensitivity and specificity of the method, calculated for different cut-off values is estimated of 60-90%, hence, is around 10% lower than the same parameters of electrophysiological tests. Difficulties in standardisation of cross-sectional nerve area (considered a classical parameter) motivated investigators to searching other indicators of the nerve compression, e.g. "wrist forearm" ratio, which is a quotient of the cross sectional area of the median nerve at the carpal tunnel inlet and 12-15 proximally at the forearm level. Some studies showed greater accuracy of this ratio, allowing to obtain the sensitivity and specificity of more than 95%. After review of the studies, authors critically conclude that actual state of art does not justify considering ultrasonography a valuable additional test in diagnosing carpal tunnel syndrome and for routine use this technique in typical cases. Ultrasonography may be useful in patients with doubtful clinical picture, as a screening test, as well as in suspicion of intra tunnel pathology. However, in atypical clinical situation, nerve conduction studies provide significantly more information on the function of the median nerve, presence of more than one compression sites or other pathology. PMID- 21648160 TI - [Swing-through gait from the perspective of biomechanics and kinesiology. Critical analysis of the current state of knowledge and the idea behind the research]. AB - The study defines the idea behind the research project which analyzes the swing through gait from the biomechanical and kinesiological perspective. In the preliminary phase, the authors performed a synthetic analysis of the state of knowledge, created a description of the general kinematic structure of the swing through gait as a form of locomotion with the use of crutches, proposed definitions. The problem was described with the use of time characteristics of vertical and horizontal anterior-posterior as well as lateral components of ground reaction forces, measured from under the supporting limb and the crutches. Presenting the idea behind the research project, the authors defined in detail the purpose of the study, the assumptions, research methodology--including a description of methods used and the measurement channels which consisted of: 2 AMTI force platforms integrated into a measurement walkway, a set of 6 optoelectronic cameras of the BTS System as well as a multichannel kinesiologic electromyography performed with the use of the NORAXON System. All phases of the research were characterized, presenting the research protocol in its entirety. The research will be conducted in the Laboratory of the Department of Biomechanics of USPS in Poznan, which possesses the ISO 9001:2008 quality management system certificate. PMID- 21648161 TI - [Patient controlled sedation]. AB - Adequate sedation is essential for successful performance of painful and uncomfortable interventional radiological, endoscopic or oncological procedures. Mild sedation is usually provided by the interventionists but an anaesthetic team is needed for deep sedation. A new alternative for intravenous sedation provided by a physician is patient controlled sedation (PCS). In PCS the patient can administer him- or herself small amounts of propofol or its combination to opioid. Spontaneous ventilation is maintained and communication with the patient is most often possible. PCS needs to be monitored adequately by an anaesthetic nurse. The advantages of PCS are appropriate level of sedation and rapid recovery. PCS has proved to be safe and effective during the change of burn injury dressings, endoscopic and radiological procedures. In Helsinki University Central Hospital the preliminary experience in over 1000 patients in the endoscopic unit has been very encouraging. PMID- 21648162 TI - [Maintenance therapy in bipolar affective disorder]. AB - In bipolar affective disorder, the patients exhibit life-long susceptibility to periodic episodes of depression, mania, hypomania, including mixed phases. Maintenance therapy aims to prevent new episodes of affective disorder and associated self-destructive behavior, to prevent milder symptoms occurring between the actual episodes and to maintain functional capacity. Maintenance therapy is always initiated upon the diagnosis of bipolar affective disorder. In a type II disorder with mild symptoms, gradual termination of maintenance therapy can be considered while monitoring the status of the patient. The drug is chosen on the basis of individual benefit/risk ratio. PMID- 21648163 TI - [Indirect comparison and network meta-analyses--new tools for the assessment of evidence on the relative efficacy of drugs]. AB - Meta-analysis allows the quantitative combination of results of multiple studies that address similar research questions. Traditional meta-analysis of studies involving a direct comparison of two treatment alternatives can be applied to estimate the overall relative efficacy of these two treatment alternatives. All treatment options relevant to practical treatment decisions are, however, not always compared directly against each other in clinical studies, but indirect comparison via a common comparator may be possible. To use all relevant evidence from both direct and indirect comparisons of treatment options, advanced methods of meta-analysis have been developed. These so-called network meta-analyses extend the traditional meta-analysis to cases where a network of studies enables different pair-wise direct and indirect comparisons between multiple treatment alternatives, thereby forming a network of relevant evidence. PMID- 21648164 TI - [Oxytocin, a neuropeptide regulating affection and social behavior]. AB - Oxytocin has been reported to modulate human behavior in many social interactions of which attachment between mother and child is the child's first social relationship. Quality of the attachment might affect the sensitivity of child's stress regulation system by modulating the development of the child's oxytocin system. This might affect social behavior in adulthood and could be one of the risk or resilience factors for mental disorders and coronary artery diseases. Oxytocin has been linked to conditions like autism and depression that significantly disturb social interaction capabilities and coping with daily life activities. Better treatment and preventative methods might be found to those diseases by investigating the effects of oxytocin to the formation of the mother child attachment and child's development. PMID- 21648165 TI - [New diagnostic criteria for pediatric osteoporosis--spinal compression fractures are an underdiagnosed problem]. AB - Osteoporosis may be associated with many pediatric conditions. Chronically ill children constitute a special risk group. Diagnosis of osteoporosis in children is challenging; contrary to adults, the diagnosis requires proof of susceptibility to fractures in addition to low bone mineral density. Vertebral fractures without high-energy injury in children are usually indicative of increased bone fragility. In chronically ill children, these fractures are surprisingly common and often asymptomatic. PMID- 21648166 TI - [Patient with high fever--the possibility of heatstroke should be kept in mind also in Finland]. AB - In heatstroke, excessive exposure to heat causes the rise of core body temperature above 40 degrees C. A disturbed state of the central nervous system is regarded as another diagnostic criterion, varying from mild disorientation to coma. The recognition of this rare state is important also in Finland, since as many as 50 to 70% of cases are fatal. In addition to direct cellular damage caused by elevated temperature, an inflammation arising in the body will in the worst case lead to endothelial injury, microthromboses and eventually to multiorgan failure. We present a fatal case of heatstroke. PMID- 21648167 TI - [Implementation of preoperative physiotherapy and recovery after shoulder impingement surgery]. AB - BACKGROUND: Subacromial impingement syndrome is a common orthopedic condition. METHODS: Patients operated due to shoulder impingement (n=104) were sent a questionnaire inquiring rehabilitation procedures before and after surgery, pain and shoulder function. RESULTS: Response rate was 68%, mean age of the patients was 53 years. Before surgery, 50% of the patients had received physiotherapy. The symptoms disappeared after surgery in the majority of the patients. However, 13% of the patients had significant shoulder pain (VAS over 30 mm) one year postoperatively and 15% had considerable functional deficit. CONCLUSIONS: To avoid unnecessary operations for shoulder impingement proper conservative treatment must be provided before surgery. PMID- 21648168 TI - [Update on current care guidelines: treatment of Crohn's disease]. AB - The incidence of Crohn's disease in Finland is 9/100,000/year and prevalence 120 200/100,000. Diagnosis is confirmed through a combination of clinical evaluation, alongside endoscopic and histological investigations. Ileocolonoscopy is the first-line diagnostic method. Radiation exposure should be considered when selecting the examination type; capsule endoscopy and MRI are therefore preferred for diagnosing diseases of the small bowel. Faecal calprotectin can be used to assess the disease's level of activity. Upon diagnosis, the course of Crohn's disease can be predicted by clinical factors (including young age, perianal disease and the initial need for corticosteroids). Early immunomodulative therapy can prevent complications, hospitalisation and operations. PMID- 21648169 TI - [Large scale questionnaire surveillance concerning invasive infections group C and G Streptococci]. PMID- 21648170 TI - Study on 99mTc-MAG3 and 99mTc-DMSA renal accumulation using in vitro cellular model. AB - Mercaptoacetyltriglycine (MAG3) and dimercaptosuccinic acid (DMSA) labelled with technetium-99m belongs to standard renal radiodiagnostics. However, the renal transport mechanisms responsible for their high renal uptake have not been fully explained. In addition, no in vitro experimental study comparing the renal uptake of these radiopharmaceuticals at the cellular level has not been performed. The investigation compared the 99mTc-MAG3 and 99mTc-DMSA renal uptake using primary rat renal cells and evaluated contribution of active and passive transport processes to the renal accumulation. The renal cells were isolated from the rat kidneys by means of the two-phase collagenase perfusion method. The used experimental model showed to be useful tool for such type of investigation. The results documented significant quantitative and qualitative differences in the accumulation of 99mTc-DMSA and 99mTc-MAG3 in the rat isolated cells. The found experimental data indicated several times higher uptake of 99mTc-MAG3 than that found in 99mTc-DMSA. 99mTc-MAG3 cellular uptake was substantially decreased when active, energy-dependent processes were inhibited. However, 99mTc-DMSA accumulation in the renal cells demonstrated only a minor dependency on energy. These findings demonstrate a very different character of the membrane transport determining 99mTc-DMSA and 99mTc-MAG3 renal accumulation. PMID- 21648172 TI - The time factor. PMID- 21648171 TI - Energy evaluation of the compaction process of directly compressible isomalt. AB - The paper compares the compressibility of two directly compressible isomalts, galenIQ 720 and galenIQ721, using the energy evaluation of the compaction process by means of the force--displacement profiles. It evaluates the energies for friction, energies accumulated by the tablet, energy of decompression, energy of compaction and plasticity in pure dry binders, in dry binders with lubricants (0.5 and 1% of magnesium stearate and sodium stearyl fumarate) and further in the tableting materials containing the model ingredients acetylsalicylic acid and ascorbic acid. The results of the study have revealed that lower values of the energy for friction and compaction with the identical compression force are found by the substance galenIQ 720, which is therefore better compressible than the substance galenIQ 721. PMID- 21648173 TI - Prucalopride. In chronic constipation: poorly documented risks. AB - Constipation is a frequent complaint, especially in women and the elderly. It is sometimes drug-induced, and is only occasionally secondary to a functional or organic disorder. The risks associated with constipation are often overestimated. Prucalopride, a 5-HT4 serotonin receptor agonist, chemically related to some neuroleptics, has been authorised in the European Union for symptomatic treatment of chronic constipation in women dissatisfied with laxatives. A combined analysis of 3 randomised double-blind trials in a total of 1999 patients (87.9% women) complaining of chronic constipation showed that about 36% of women considered it effective at a dose of 2 or 4 mg/day, versus 18% of women receiving placebo. Normal bowel movements resumed in respectively 23.6% and 24.7% of patients taking 2 and 4 mg/day prucalopride, versus 11.3% of patients on placebo (p < 0.001). No statistically significant difference was found between the 2 doses of prucalopride. Palpitations were more frequent in patients treated with prucalopride. The incidence of ischaemic cardiovascular events was 0.2% with prucalopride versus 0.1% with placebo. Increases in heart rate and blood pressure were observed in pigs and dogs treated with prucalopride. Prucalopride seems to increase prolactin levels. Tumours of the liver and thyroid were observed in rats. Prucalopride also carries a risk of poorly defined pharmacokinetic and pharmacodynamic interactions. Prucalopride may reduce the efficacy of oral contraceptives. Miscarriages were reported in clinical trials. Prucalopride should not be taken during pregnancy. In addition, all women of child-bearing age should use effective contraception while taking prucalopride. In practice, prucalopride should be avoided. It is better to focus on lifestyle and behavioural changes, and rational use of laxatives. PMID- 21648174 TI - INN common stem: -prost-. PMID- 21648175 TI - Quadrivalent papillomavirus vaccine in women over age 26. Cervical screening must remain a priority. PMID- 21648176 TI - Autologous chondrocytes. Autologous chondrocyte implantation: more data needed. AB - There is no standard surgical treatment for young adults with persistent, incapacitating symptoms of knee cartilage damage. ChondroCelect is the first cell therapy product to be authorised in the European Union. It contains a dense suspension of chondrocytes cultured from a biopsy of the patient's knee cartilage for 4 weeks before being reimplanted. Clinical evaluation of Chondro-Celect only includes one trial, versus subchondral microfracture, in 118 patients. After 3 years of follow-up, there was no difference in the symptom score between the groups. Histological outcome was better after autologous chondrocyte implantation, but methodological problems make it difficult to interpret the observed difference. Long-term functional outcomes remain to be determined. More joint complications occurred after autologous chondrocyte implantation than after subchondral bone microfracture: more frequently symptomatic cartilage hypertrophy (27% versus 13%, possibly related to the implantation technique), joint swelling (22% versus 6.6%), joint effusion (24% versus 9.8%), and joint crepitations (18% versus 6.6%). Autologous chondrocyte implantation was sometimes associated with flu-like syndrome (in 7.8% of patients), which did not occur with the microfracture technique. Autologous chondrocyte implantation is more complex than microfracture. During routine use, there is a risk that one patient will inadvertently receive chondrocytes collected from another patient, leading to a risk of rejection. In practice, this autologous chondrocyte product should only be used by highly specialised teams, and its assessment must continue. PMID- 21648177 TI - Nimesulide: patients still exposed to a risk of severe hepatitis. AB - Nimesulide, a nonsteroidal antiinflammatory drug (NSAID) that has been marketed in France since 1998, is neither more effective nor better tolerated than other NSAIDs. Many reports and reviews published by drug regulatory agencies in Spain, Ireland and Italy have warned of the hepatic adverse effects of nimesulide. In early 2008, 17 cases of nimesulide-induced liver damage requiring transplantation had been reported in Ireland, Italy, Spain, Finland and France. An Italian retrospective study conducted between 1997 and 2001, including about 400 000 patients exposed to NSAIDs, showed that the risk of severe liver damage was twice as high with nimesulide as with other NSAIDs. The European pharmacovigilance database shows that nimesulide is associated with more cases of severe liver damage than other NSAIDs, as well as more cases of liver damage than with cox-2 inhibitors. Young women are particularly at risk. In the vast majority of cases of liver damage, the dose of nimesulide used was that recommended in the summary of product characteristics (SPC). Liver damage occurred within 15 days after taking the first dose of nimesulide in one-third of cases. It is unacceptable that nimesulide still remains on the market in France and other countries: reports of severe liver damage continue to accumulate, and many other therapeutic options are available. PMID- 21648178 TI - Azathioprine and mercaptopurine: lymphoma. AB - Prospective follow-up for a median of 35 months of a French cohort of 19 486 patients with inflammatory bowel disease showed a nearly 4-fold increase in the risk of lymphoma in patients exposed to azathioprine or mercaptopurine (relative risk 3.75; 95% confidence interval 1.59 to 8.85). This risk should be taken into account when weighing the likely benefits of these drugs in treatment of patients with severe chronic inflammatory bowel disease no longer responding to first-line treatment. PMID- 21648179 TI - Poppers: methaemoglobinaemia; respiratory, cardiac and visual disorders. PMID- 21648180 TI - Treatment-resistant depression: no panacea, many uncertainties. Adverse effects are a major factor in treatment choice. AB - At least 50% of patients with depression do not enter remission after several weeks of antidepressant therapy. To determine the treatment options and their respective risk-benefit balances in this setting, we reviewed the literature using the standard Prescrire methodology. Clinical trials and epidemiological studies show that depression should only be considered drug-resistant after at least 6 weeks of therapy. After assessing residual symptoms and their impact on the patient's quality of life, a search should be made for factors responsible for the persistence of depression, such as the patient's environment, a psychiatric or somatic disorder, and drug intake or addiction. Increasing the dose of the first-line antidepressant is only based on weak evidence. Trials comparing continuing the first-line antidepressant versus switching to another pharmacological class have yielded conflicting results. A switch may benefit some patients, but the elimination half-life of the discontinued drug must be taken into account to limit the risk of interactions during the transition. Combining two antidepressants mainly increases the risk of adverse effects, without a tangible clinical benefit. Two meta-analyses suggest that adding a so-called atypical neuroleptic to ongoing antidepressant therapy leads to 1 extra remission per 7 to 10 treated patients, but also to treatment cessation due to adverse effects in 8% to 9% of cases. Older neuroleptics have not been properly evaluated in this setting. Comparative trials suggest that lithium may have a certain antidepressant effect in this setting, but there is no firm evidence that adding lithium increases the chances of remission. Lithium has a narrow therapeutic margin and overdose can be fatal; the blood lithium concentration must therefore be monitored. Adding an antiepileptic or a psychostimulant is more harmful than beneficial. Adding a thyroid hormone, a benzodiazepine, buspirone or pindolol has no proven antidepressive effect. Four trials, each including fewer than 20 patients, have assessed the efficacy of psychotherapy in patients with treatment resistant depression. Two of them provided positive results. Electroconvulsive therapy is probably effective for some patients with refractory depression but it necessitates general anaesthesia and carries a risk of memory disorders. Vagal nerve electrostimulation has no proven efficacy. Transcranial magnetic stimulation seems to have some efficacy and few adverse effects, but its optimal modalities remain to be determined. In practice, when the patient and doctor decide to attempt second-line therapy for treatment-resistant depression, adverse effects must be taken into account in the choice of drug(s). Maintaining a good quality relationship between patient and doctor may be more important than attempting to obtain remission "at any cost". PMID- 21648181 TI - The finances of Association Mieux Prescrire. 2010 Prescrire annual report. PMID- 21648183 TI - Patient education: keep the best, avoid the rest. PMID- 21648182 TI - Prescrire's proposals to France's national conference on medicines policy: patients' interests and public health should come first. PMID- 21648184 TI - Neuroendocrine activation as a target of modern chronic heart failure pharmacotherapy. AB - At present, a constant progress in pathophysiology understanding and treatment of the chronic heart failure (CHF) is arising. The current CHF pharmacotherapy is complex, involving factors affecting the renin-angiotensin-aldosterone system (RAAS), beta-blockers, diuretics and vasodilatators. There are also significant efforts to introduce in CHF pharmacology novel therapeutic strategies, based on the other neurohormonal mechanisms activated in CHF. They include vasopressin receptor antagonists (VRA; vaptans), endothelin receptor antagonists (ERA; sentans), agents relating to the natriuretic peptides system (neutral endopeptidase inhibitors; NEPI and vasopeptidase inhibitors; VPI) and anticytokines agents (anti TNF-alpha immunoglobulin or TNF-alpha scavenger receptor; Etanercept). In this article we briefly describe the modem approach to CHF systemic treatment. PMID- 21648185 TI - Comparison of classic and derivative UV spectrophotometric methods for quantification of meloxicam and mefenamic acid in pharmaceutical preparations. AB - The methods for quantitative determination of meloxicam and mefenamic acid in pharmaceuticals by classic spectrophotometry - zero order derivative, first and second order derivatives spectrophotometry is described, using "peak - peak" (P P) and "peak - zero" (P-O) measurements. The calibration curves are linear within the concentration range of 4.0 - 14.0 microg/mL for meloxicam and 14.0 - 24.0 microg/mL for mefenamic acid. The procedure is simple, rapid and the results are reliable. PMID- 21648186 TI - Determination of carvedilol by its quenching effect on the luminescence of terbium complex in dosage form. AB - A new, simple, sensitive luminescence methods for the determination of carvedilol have been developed and validated. Carvedilol was remarkably quenching the luminescence intensity of the Tb(III) ion in the new terbium complex with 1-butyl 4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxylic acid-(4-methyl-pyridin-2-yl) amide (R) in aqueous solutions containing urotropine buffer (pH 7.5) at lambda(ex) = 317 nm and lambda(em) = 545 nm. Under optimal conditions, the quenching of luminescence intensity was found to be proportional to the concentration of carvedilol in the range of 0.5-400 microg/mL. The detection limit was 0.16 microg/mL. This method was applied for the determination of carvedilol in tablets "Coryol". PMID- 21648187 TI - Combination treatment with 5-fluorouracil and isothiocyanates shows an antagonistic effect in Chinese hamster fibroblast cells line-V79. AB - The isothiocyanates present in the cruciferous plants were proved to have the antiproliferative and cytotoxic effect on cancer cell lines. Natural compounds in combination with chemotherapy agents enhance anticancer activities of drugs and reduce their toxicity. The aim of the presented study was to determine an effect of isothiocyanates and 5-fluorouracil used alone or in combination (in sequential or co-administrative treatments) on normal cell lines-V79. There were compared abilities of three isothiocyanates to interact with 5-fluorouracil. There was also investigated the mechanism of interaction and influence of isothiocyanates on 5-fluorouracil. The cell survival was evaluated with MTT assay. Combination effects between isothiocyanates and 5-fluorouracil were estimated in the way described by Chou and Talalay. The cycle progression and the living cells number were determined with flow cytometry. The type of cell death was detected with a confocal microscope. There was observed an antagonistic effect which was mainly dependent on the cell cycle distribution e.g., sulforaphane increased the cell number in the G2/M phase, whereas 5-fluorouracil and combination of these two compounds increased the cell number in the S phase. If each compound blocked the S phase of the cell cycle, their combination increased the cell number in the S phase, but the increase was not statistically significant when compared with single substance treatments. The highest antagonistic effect in normal cells was obtained for co-administrated 5-fluorouracil and 2-oxoheptyl isothiocyanate at the fraction affected at 0.5 and 0.75. Isothiocyanates did not affect 5 fluorouracil cytotoxicity in normal cell lines-V79. PMID- 21648188 TI - Synthesis and anti-HIV activity of novel 3-substituted phenyl-6,7-dimethoxy-3a,4 dihydro-3H-indeno[1,2-c]isoxazole analogues. AB - A series of novel 3-(substituted phenyl)-6,7-dimethoxy-3a,4-dihydro-3H-indeno[1,2 c]isoxazole analogues were synthesized by the reaction of 5,6-dimethoxy-2-[(E)-1 phenylmethylidene]-1-indanone with hydroxylamine hydrochloride. The title compounds were tested for their in vitro anti-HIV activity. Among the compounds, (4g) showed a promising anti-HIV activity in the in vitro testing against IIIB and ROD strains. The IC50 of both IIIB and ROD were found to be 9.05 microM and > 125 microM, respectively. PMID- 21648189 TI - Synthesis and antiproliferative activity in vitro of new pyrido[1,4-b]diazepine derivatives and imidazo[4,5-b]pyridine. AB - A novel series of esters 8-10 and hydrazones 4-6 was synthesized from 4-aryl-2 phenacylidene-1,3,4,5-tetrahydropyrido[2,3-b][1,4]diazepine (1-3). Subsequent treatment of hydrazone 4 with p-chlorbenzaldehyde furnished azine 7. Long standing heating of ester 8 with hydrazine hydrate afforded 3-[1-(p chlorophenylene)-2-(5-phenyl-1H-pyrazol-3-yl)-ethyl]-1,3-dihydroimidazo[4,5 b]pyridin-2-one (11). The structures of 4-6 and 8-10 were identified by the results of elemental analysis and their IR, 1H-NMR and MS spectra. Additionally, the structure of 11 was confirmed by X-ray diffraction method. Compounds 8-10 and 11 were examined for their antiproliferative activity in vitro against the cells of 5 human cancer cell lines, using SRB or MTT technique. Among tested compounds, only 11 revealed cytotoxic activity in vitro against all cell lines applied with ID50 (inhibitory dose 50%) values lower than 4 microg/mL, which is an international activity criterion for synthetic compounds. All compounds inhibit the proliferation of HL-60 human promyelocytic leukemia cell line. PMID- 21648190 TI - Synthesis, antitumor activity and molecular docking study of novel benzofuran-2 yl pyrazole pyrimidine derivatives. AB - A new series of (benzofuran-2-yl)-1-phenyl-1H-pyrazol-4-yl) pyrimidine derivatives were synthesized from 3-(benzofuran-2-yl)-1-phenyl-1H-pyrazol-4 carbaldehyde (1) through different routes of cyclocondensation reactions. Condensation of 1 with active methylene compounds afforded compounds 2-8. The cyclization of 2 with chloroacetic acid, ortho substituted benzoic acid and/or ethanolamine gave compounds 9-12. Also condensation of 2 with hydrazine hydrate followed by cyclocondensation afforded corresponding triazines and pyrazole derivatives 18-27. Some docking studies of the newly prepared compounds as thymidylate synthase inhibitors have been done. Also the cytotoxic activity of some of the prepared compounds as a representative examples was evaluated against HEPG2 (human liver carcinoma cell line) in comparison with 5-fluorouracil (5-Fu). PMID- 21648191 TI - Synthesis, characterization and in vivo anticonvulsant and neurotoxicity screening of Schiff bases of phthalimide. AB - A series of Schiff bases of phthalimide (4a-l) were prepared in satisfactory yields and evaluated for their anticonvulsant and neurotoxicity activities. The structures of all the compounds were in good agreement with elemental analysis and spectral data. All the compounds were active in MES screen and less neurotoxic than phenytoin. Compound 41 having nitro substitution at ortho position of the distal aryl ring emerged as most promising anticonvulsant agent with low neurotoxicity. PMID- 21648192 TI - Synthesis of some new 2-(substituted-phenyl)-5-(N,N-diphenylaminomethyl)-1,3,4 oxadiazoles: a safer anti-inflammatory and analgesic agents. AB - A series of 2-(substituted-phenyl)-5-(N,N-diphenylaminomethyl)-1,3,4-oxadiazoles (3-15) were synthesized. The compounds were evaluated for their anti inflammatory, analgesic, ulcerogenic and lipid peroxidation actions. The percentage inhibition in edema at different time intervals indicated that compounds 8, 11, 12, 14 and 15 exhibited good anti-inflammatory potential. The results illustrate that 2-(2-acetoxyphenyl)-5-(N,N-diphenylaminomethyl)-1,3,4 oxadiazole (15) and 2-(3,4-dimethoxyphenyl)-5-(N,N-diphenylaminomethyl)-1,3,4 oxadiazole (12) showed best anti-inflammatory activity among the series tested. Furthermore, activity is higher in case of chloro substitution as compared to methyl substitution. The compounds synthesized were also evaluated for their ulcerogenic and lipid peroxidation action and showed superior GI safety profile along with reduction in lipid peroxidation as compared to that of ibuprofen. PMID- 21648193 TI - Synthesis of P-triazinylphosphonium salts--hybrid molecules with potential antimicrobial activity. AB - The new hybrid drugs combining in a single molecule triazine ring attached to phosphonium salt were prepared and their bactericidal activity against Gram positive bacteria Staphylococcus aureus ATCC43300 (MRSA - methycyline resistant Staphylococcus aureus strain) two Gram-negative bacteria Escherichia coli CCUG31997 serotype O153 (EPEC - enteropathogenic Escherichia coli strain), Proteus mirabilis 1784 (MDR - multidrug resistant clinical strain) were determined using microdilution method. PMID- 21648194 TI - New steroidal lactones and homomonoterpenic glucoside from fruits of Malva sylvestris L. AB - Phytochemical investigation of the ethanolic extract of defatted fruits of Malva sylvestris Linn. (Malvaceae) led to the isolation of six new steroidal lactones and a homomonoterpenic glucoside along with beta-sitosterol-3-beta-D glucopyranoside. The structures of new phytoconstituents have been elucidated as cholest-5-en-3a-ol-18(21)-olide (sylvestrosterol A), cholest-9(11)-en-3alpha-ol 18(21)-olide (sylvestrosterol B), cholest-4,6,22-trien-3alpha-ol-18(21)-olide (sylvestrosterol C), 2-methyl-6-methylene-n-decan-2-olyl- 3beta-D-glucopyranoside (malvanoyl glucoside), cholest-7-en-18(21)-olide-3alpha-olyl-3beta-D glucopyranoside (sylvestrogenin A), cholest-9(11)-en-18(21)-olide-3alpha-olyl 3beta-D-glucopyranoside (sylvestrogenin B) and cholest-5-en-8(21)-olide-3alpha olyl-3beta-D-glucopyranoside (sylvestrogenin C).The structures of all these phytoconstituents have been established on the basis of spectral data analysis and chemical reactions. PMID- 21648195 TI - Flavonoids from the flowers of Aesculus hippocastanum. AB - The flavonoids, kaempferol derivatives: 3-O-alpha-arabinofuranoside, 3-O-beta glucopyranoside, 3-O-alpha-rhamnopyranoside, 3-O-alpha-rhamnopyranosyl (1 --> 6) O-beta-glucopyranoside and quercetin derivatives: 3-O-alpha-arabinofuranoside, 3 O-beta-glucopyranoside, 3-O-alpha-rhamnopyranosyl (1 --> 6)-O-beta glucopyranoside, were isolated from the flowers of Aesculus hippocastanum and identified. The structures of these compounds were confirmed by a chemical analysis and spectrophotometric methods (UV, 1H-, 13C-NMR, ESI-MS). The presence of free aglycones: kaempferol and quercetin was confirmed chromatographically by comparison with standards. PMID- 21648196 TI - Preparation and in vitro dissolution profile of zidovudine loaded microspheres made of Eudragit RS 100, RL 100 and their combinations. AB - The objective of present investigation was to evaluate the entrapment efficiency of the anti-HIV drug, zidovudine, using two Eudragit polymers of different permeability characteristics and to study the effect of this entrapment on the drug release properties. In order to increase the entrapment efficiency optimum concentration of polymer solutions were prepared in acetone using magnesium stearate as droplet stabilizer. The morphology of the microspheres was evaluated using a scanning electron microscope, which showed a spherical shape with smooth surface. The mean sphere diameter was between 1000-3000 microm and the entrapment efficiencies ranged from 56.4-87.1%. Polymers were used separately and in combination to prepare different microspheres. The prepared microspheres were studied for drug release behavior in phosphate buffer at pH 7.4, because the Eudragit polymers are independent of the pH of the dissolution medium. The release profiles and entrapment efficiencies depended strongly on the structure of the polymers used as wall materials. The release rate of zidovudine from Eudragit RS 100 microspheres was much lower than that from Eudragit RL 100 microspheres. Evaluation of release data reveals that release of zidovudine from Eudragit RL 100 microspheres followed the Higuchi rule, whereas Eudragit RS 100 microspheres exhibited an initial burst release, a lag period for entry of surrounding dissolution medium into polymer matrix and finally, diffusion of drug through the wall material. PMID- 21648197 TI - Alternative and generalized approach to in vitro-in vivo correlation. AB - Evaluation of in vitro-in vivo correlation (IVIVC) plays important role in securing therapeutic effect if a dosage form undergoes technological modifications. Similarity (closeness) of dissolution profiles of the original and modified dosage forms has been traditionally considered to be sufficient for similar in vivo responses. This may be true if the IVIVC model (dependence between the dissolution and corresponding absorption profiles) is given by a linear straight line with the unit slope. The paper presents an alternative and generalized approach to IVIVC evaluation. Influences of pre-systemic processes (disintegration, dissolution, absorption) on the system response (concentration time profile C(t), bioavailability BD and other) are analyzed and evaluated. Both the magnitude and sign of IVIVC are then derived from the magnitudes and signs of these influences. The underlining idea is that pre-systemic processes do not correlate with the system response, (e.g., plasmatic concentration) if small modifications of the former do not induce significant changes of the later. If this is so, the therapeutic effects of the modified and original dosage forms may be considered equal or at least similar. In this way the problem of IVIVC is not only exactly mathematically founded but modifications of pre-systemic processes are directly projected to the system output-- the time profile of plasmatic concentration. Moreover, the approach is applicable to virtually any dosage form. Its feasibility was validated in vivo. PMID- 21648198 TI - Evaluation of drug-excipient interaction in the formulation of celecoxib tablets. AB - In the present study, the possible interactions between celecoxib and some excipients (colloidal silicon dioxide (Aerosil), microcrystalline cellulose (Avicel PH 102), lactose anhydrous, magnesium stearate, cross-povidone and talc) were evaluated by examining the pure drug or drug-excipient powder mixtures which were stored under different conditions (25 +/- 2 degrees C, 60% RH +/- 5% RH or 40 + 2 degrees C, 75% RH +/- 5% RH) and different period (30 or 60 days) using DSC, FT-IR and HPLC. In order to investigate the possibility of celecoxib excipient interaction in aqueous medium, dispersions of the pure drug or drug in physical powder mixture (1:1 w/w) in water (1%, w/v) were also prepared and evaluated by FT-IR and HPLC at day 0 and day 7 (40 +/- 2 degrees C). The interaction between celecoxib and magnesium stearate or colloidal silicon dioxide were determined in the aqueous dispersions by FT-IR. Different tablet formulations with or without excipients tested were prepared, and assessed for drug dissolution and permeability. PMID- 21648199 TI - Combination of aripiprazole and ethanol attenuates marble-burying behavior in mice. AB - Obsessive-Compulsive Disorder (OCD) is characterized by absurd, recurrent thoughts (obsessions) followed by certain stereotyped actions (compulsions). 5 Hydroxytryptamine (5-HT) abnormalities may be involved in OCD, and further, cause changes in serotonergic transmission that may have direct or indirect effects on the neuronal firing of other neuromodulators affecting thoughts, feelings and behaviors. Serotonin-related genes that are found in OCD include those coding for the 5-HT transporter (5-HTT) and receptors (5-HT(2A), 5-HT(2B), 5-HT(2C) and 5 HT(1B)) as well the 5-HT enzyme tryptophan hydroxylase. OCD can impair all areas of brain functioning and produce devastating effects on patients and their families. Marble-burying behavior of mice has been employed to study anxiety disorders, including the OCD. The aim of this study was to test the efficacy of aripiprazole and alcohol per se and in combination on marble-burying behavior of mice. A total of 114 male Swiss mice divided in 19 groups were studied. Aripiprazole (0.1 mg/kg, i.p.) per se as well as ethanol (0.1% w/v) per se did not show any anti-compulsive activity. But the combination comprising of ineffective doses of aripiprazole (0.1 mg/kg, i.p.) and ethanol (0.1% w/v) showed significant anti-compulsive activity as reflected by inhibition of marble-burying behavior. PMID- 21648200 TI - Synthesis of biologically active heterocycles from coumarin thiocyanate. PMID- 21648201 TI - QSAR studies on human 5alpha-reductase inhibitors: unsaturated 3-carboxysteroids. PMID- 21648202 TI - Spectrofluorimetric estimation of puerarin in Pueraria tuberosa. PMID- 21648203 TI - Part 1. Short-term effects of air pollution on mortality: results from a time series analysis in Chennai, India. AB - This report describes the results of a time-series analysis of the effect of short-term exposure to particulate matter with an aerodynamic diameter < or = 10 pm (PM10) on mortality in metropolitan Chennai, India (formerly Madras). This was one of three sites in India chosen by HEI as part of its Public Health and Air Pollution in Asia (PAPA) initiative. The study involved integration and analysis of retrospective data for the years 2002 through 2004. The data were obtained from relevant government agencies in charge of routine data collection. Data on meteorologic confounders (including temperature, relative humidity, and dew point) were available on all days of the study period. Data on mortality were also available on all days, but information on cause-of-death (including accidental deaths) could not be reliably ascertained. Hence, only all-cause daily mortality was used as the major outcome for the time-series analyses. Data on PM10, nitrogen dioxide (NO2), and sulfur dioxide (SO2) were limited to a much smaller number of days, but spanned the full study period. Data limitations resulting from low sensitivity of gaseous pollutant measurements led to using only PM10 in the main analysis. Of the eight operational ambient air quality monitor (AQM) stations in the city, seven met the selection criteria set forth in the common protocol developed for the three PAPA studies in India. In addition, all raw data used in the analysis were subjected to additional quality assurance (QA) and quality control (QC) criteria to ensure the validity of the measurements. Two salient features of the PM10 data set in Chennai were a high percentage of missing readings and a low correlation among daily data recorded by the AQMs. The latter resulted partly because each AQM had a small footprint (approximate area over which the air pollutant measurements recorded in the AQM are considered valid), and partly because of differences in source profiles among the 10 zones within the city. The zones were defined by the Chennai Corporation based on population density. Alternative exposure series were developed to control for these data features. We first developed exposure series based on data from single AQMs and multiple AQMs. Because neither was found to satisfactorily represent population exposures, we subsequently developed an exposure series that disaggregated pollutant data to individual zones within the city boundary. The zonal series, despite some uncertainties, was found to best represent population exposures among other available choices. The core model was thus a zonal model developed using disaggregated mortality and pollutant data from individual zones. We used quasi-Poisson generalized additive models (GAMs) with smooth functions of time, temperature, and relative humidity modeled using penalized splines. The degrees of freedom (df) for these confounders were selected to maximize the precision with which the relative risk for PM10 was estimated. This is a deviation from the traditional approaches to degrees of freedom selection, which usually aim to optimize overall model fit. Our approach led to the use of 8 df/year for time, 6 df/year for temperature, and 5 df/year for relative humidity. The core model estimated a 0.44% (95% confidence interval [CI] = 0.17 to 0.71) increase in daily all-cause mortality per 10-pg/m3 increase in daily average PM10 concentrations. Extensive sensitivity analyses compared models constructed using alternative exposure series and contributions of model parameters to the core model with regard to confounder degrees of freedom, alternative lags for exposure and meteorologic confounders, inclusion of outliers, seasonality, inclusion of multiple pollutants, and stratification by sex and age. The sensitivity analyses showed that our estimates were robust to a range of specifications and were also comparable to estimates reported in previous time-series studies: PAPA, the National Morbidity, Mortality, and Air Pollution Study (NMMAPS), Air Pollution and Health: A European Approach (APHEA), and Air Pollution and Health: A European and North American Approach (APHENA). While the approaches developed in previous studies served as the basis for our model development, the present study has new refinements that have allowed us to address specific data limitations (such as missing measurements and small footprints of air pollution monitors). The methods developed in the study may allow better use of routine data for time-series analysis in a broad range of settings where similar exposure and data-related issues prevail. We hope that the estimates derived in this study, although somewhat tentative, will facilitate local environmental management initiatives and spur future studies. PMID- 21648204 TI - Part 2. Time-series study on air pollution and mortality in Delhi. AB - INTRODUCTION: Air pollution concentrations in most of the megacities in India exceed the air quality guidelines recommended by the World Health Organization and may adversely affect human health in these cities. Particulate matter (PM) is the pollutant of concern in many Indian cities, particularly in the capital city of Delhi, In recent years, several actions have been taken to address the growing air pollution problem in Delhi and other Indian cities; however, few studies have been designed to assess the health effects of air pollution in Indian cities. To bridge the gap in scientific knowledge and add evidence to the ongoing studies in other Asian cities, a retrospective time-series study on air pollution and mortality in Delhi was initiated under the HEI Public Health and Air Pollution in Asia (PAPA) program. APPROACH: The study used retrospective time-series data of air quality and of naturally-occurring deaths recorded in Delhi to identify changes in the daily all-natural-cause mortality rate that could be attributed to changes in air quality. The 3-year study period included the years 2002 through 2004. The methodology involved: (1) collecting data on ambient air quality for major pollutants from all monitoring stations in Delhi; (2) collecting meteorologic data (temperature, humidity, and visibility); (3) collecting daily mortality records from the Registrar of Births and Deaths; (4) statistically analyzing the data using the common protocol for Indian PAPA studies, which included city-specific modifications. RESULTS AND IMPLICATIONS: The study findings showed that increased concentrations of PM with an aerodynamic diameter < or = 10 microg/m3 (PM10) and of nitrogen dioxide (NO2) were associated with increased all-natural-cause mortality. It was found that every 10-microg/m3 change in PM10 was associated with only a 0.15% increase in total all-natural cause mortality. When NO2 alone was considered in the model, daily all-natural cause mortality increased 0.84% for every 10-microg/m3 increase in NO2 concentration. No significant effect was observed for changes in sulfur dioxide (SO2) concentrations. The study provides insight into the link between air pollution and mortality in local populations and contributes information to the existing body of knowledge. PMID- 21648205 TI - We want our money back! PMID- 21648206 TI - Mifamurtide: osteosarcoma: ineffective and harmful. AB - The standard treatment for children and young adults with osteosarcoma consists of surgery, preceded and followed by methotrexate-based chemotherapy. Mifamurtide is an immunostimulant derived from a bacterial cell wall component. It is authorised in the European Union as an adjunct to combination chemotherapy after complete excision of non-metastatic osteosarcoma. Only one comparative, unblinded trial has been published, and its design was particularly complex. In a study population of 678 patients, adding mifamurtide to chemotherapy after tumour excision did not prolong the overall 6-year survival rate, which was about 75% with both treatments. Only serious adverse effects were collected, and they were not systematically recorded. Hypersensitivity reactions occurred in clinical trials, along with pleural and pericardial effusions, seizures, and muscle spasms. Severe hearing loss occurred in 12% of the patients treated with mifamurtide in the comparative trial, versus 7% of the other patients. In practice, given the lack of any survival benefit and the risk of serious adverse effects, it is better not to add mifamurtide to chemotherapy regimens used for treatment of osteosarcoma. PMID- 21648207 TI - Omalizumab: a second look in severe persistent asthma: new adverse effects. AB - When severe asthma persists despite high-dose inhaled corticosteroid therapy, it is better to switch to oral corticosteroid therapy than to add another asthma drug. In 2007, we concluded that omalizumab had a negative risk-benefit balance in this setting. In mid-2010, we found no new data on the efficacy of omalizumab. At best, omalizumab prevents 1 emergency room visit approximately every 2.5 years. Post-marketing follow-up data confirm the infectious adverse effects and hypersensitivity reactions associated with omalizumab, and show that anaphylactic reactions can occur several days after an injection. A possible increase in the risk of cancer has not yet been ruled out. New adverse effects associated with omalizumab have emerged, including serious cardiac effects and severe thrombocytopenia. In practice, despite longer followup, the efficacy of omalizumab remains uncertain, while new, sometimes serious adverse effects have emerged. It is better to adjust standard treatments to the individual's situation than to risk using omalizumab. PMID- 21648208 TI - Omalizumab in children: a negative risk-benefit balance in severe persistent asthma. PMID- 21648209 TI - Trabectedin and ovarian cancer: statistical trickery. PMID- 21648210 TI - Rosuvastatin and primary cardiovascular prevention. Continue to use pravastatin or simvastatin. PMID- 21648211 TI - Bradycardia due to cholinesterase inhibitors: identify adverse effects and take them into account. AB - The cholinesterase inhibitors donepezil, rivastigmine and galantamine have a modest and transient benefit in Alzheimer's disease. Their known adverse effects include bradycardia. A Canadian case-control study conducted between 2003 and 2008 showed a statistically significant increase in the risk of hospitalisation for bradycardia among patients who had been taking a cholinesterase inhibitor for less than 3 months, compared with patients who had stopped taking a cholinesterase inhibitor more than 6 months previously. After hospital discharge, more than half of these patients were again prescribed a cholinesterase inhibitor, and 4% of them were re-admitted for bradycardia. In practice, when an adverse effect has been identified and treated, this information must be shared and taken into account by all those involved in the patient's subsequent management. PMID- 21648212 TI - Bisphosphonates and atrial fibrillation: clinical trial data suggest possible link. AB - In 2007, an article describing an American placebo-controlled clinical trial of yearly zoledronic acid infusion at a dose of 5 mg mentioned a statistically significant excess of severe atrial fibrillation in postmenopausal women. Bisphosphonates had not previously been linked to this adverse effect. Several meta-analyses of clinical trials have been published, but they covered only a small number of the hundreds of comparative trials evaluating bisphosphonates, few of which mentioned atrial fibrillation. In 2010, a meta-analysis of placebo controlled trials of bisphosphonates, in a total of 26 126 patients with osteoporosis, showed an increased risk of severe atrial fibrillation. A larger meta-analysis (40104 women) showed a nonsignificant increase in the risk of atrial fibrillation, regardless of severity. A meta-analysis published in 2010 included 7 placebo-controlled observational studies of bisphosphonates in patients with osteoporosis, two of which showed an increased risk of severe atrial fibrillation. This meta-analysis showed no statistically significant increase in the overall risk of severe atrial fibrillation. Our literature search, up to mid-2010, found no studies concerning the risk of atrial fibrillation in cancer patients treated with bisphosphonates. No cardiac adverse effects were mentioned in a report analysing two trials in patients with Paget's disease. Pending the publication of more data, the potential risk of severe atrial fibrillation in some patients treated with bisphosphonates should be taken into account. There is no evidence of an increased risk with a specific bisphosphonate, route of administration, patient subpopulation, or treatment duration. PMID- 21648213 TI - Upper gastrointestinal bleeding and spironolactone. AB - Spironolactone is a potassium-sparing diuretic marketed since the 1960s that is known to cause the same adverse effects as other diuretics as well as some effects of its own, such as hyperkalaemia, gynaecomastia and menstrual disturbances. A cohort study and 2 case-control studies concurred to show an approximately twofold, dose-dependent increase in the incidence of upper gastrointestinal bleeding in patients receiving spironolactone. The postulated mechanism is that spironolactone's aldosterone antagonist activity delays gastroduodenal healing. However, gastrointestinal bleeding is not a common adverse effect of aldosterone antagonists. In practice, although these studies have a low level of evidence, the role of spironolactone should be suspected in the event of a gastrointestinal bleed and caution should be exercised with spironolactone in patients who have predisposing factors to gastrointestinal bleeding. PMID- 21648214 TI - Buflomedil: still too many fatal cardiac and neurological events. PMID- 21648215 TI - Drug interactions with cannabis. PMID- 21648216 TI - First-line antiretroviral treatment of HIV-infected children. A choice largely based on adult data. AB - About 2 million children worldwide are infected with HIV.There are about 1500 HIV infected children in France. This review examines the choice of first-line antiretroviral treatment for children under 12 years of age, based on a review of the literature using the standard Prescrire methodology. In children under 5 years of age, the percentage of CD4+ T lymphocytes among all circulating lymphocytes is more stable than the absolute count. The CD4+ T cell percentage is therefore used as a marker of immunological status in this age group. The decision to begin antiretroviral treatment in a child depends on the short or medium-term risk of progression to AIDS or death. Treatment is warranted for infants under 1 year of age, children under 6 years of age whose CD4+ T cell percentage is below 25%, and older children with a CD4+ T cell count below 350 per mm3. In Africa, antiretroviral treatment seems justified for all infected children under 2 years of age. First-line treatment for children is based on a combination of at least 3 antiretroviral drugs. Zidovudine and lamivudine remain the first-choice nucleoside reverse transcriptase inhibitors. The lopinavir + ritonavir combination is the first-choice HIV protease inhibitor, but the oral solution is poorly accepted by children because of its unpleasant taste and the high ethanol content of the ritonavir oral solution. As for the non-nucleoside reverse transcriptase inhibitors, efavirenz has not been assessed in children under 3 years of age; nevirapine can be used, but it has suboptimal antiviral activity. Antiretroviral drugs seem to have similar adverse effects in children and adults. Long-term data are lacking, however, especially on possible cardiac and metabolic effects. In view of the large number of children infected worldwide, more efforts are urgently needed to adapt first-line antiretroviral drugs to paediatric use. This means developing and assessing specific paediatric formulations and fixed-dose combinations. PMID- 21648217 TI - New drugs and indications in 2010: inadequate assessment; patients at risk. AB - In 2010, we rated 97 new drugs or new indications in our French edition la revue Prescrire, only 4 of which provided a therapeutic advantage. However, 19 others (1 in 5) were approved despite having more harms than benefits. More paediatric products were released in 2010 than in previous years, but few of them made any real difference and many had not been properly evaluated. Drug regulatory agencies can protect patients from exposure to dangerous drugs by refusing to grant market approval or by demanding their market withdrawal.Yet they are failing to fulfil this responsibility: so-called risk management plans and modifications to the wording in the SPC are only half-measures. Too often the authorities put companies' short-term financial interests above patients' well being by granting premature marketing authorisation, by agreeing to high levels of reimbursement that fail to take added therapeutic value into account, and by allowing the development of "umbrella" ranges. The European authorities' questionable plans for pharmacovigilance and advertising of prescription-only drugs were restricted after public mobilisation, but they are still likely to undermine healthcare quality. Decision-makers must make patients' well-being their top priority. PMID- 21648218 TI - Drugs to avoid. PMID- 21648219 TI - Medicines agencies too often under the influence of drug companies. PMID- 21648220 TI - Finding solutions, along with patients. PMID- 21648221 TI - [Expression and utilization of 3AB nonstructural protein of foot-and-mouth disease virus in Escherichia coli]. AB - To develop a sensitive and specific ELISA for detection of antibodies to the nonstructural protein of FMDV. We cloned and expressed FMDV nonstructural protein 3AB in Escherichia coli expression system. The recombinant protein 3AB was purified with Ni-NTA HisBind Resins and characterized by Western blotting. An indirect ELISA based on purified protein 3AB as a coating antigen was established. The specificity and sensitivity of this assay were evaluated by comparison with a commercial 3ABC-ELISA kit in detecion of serum samples. The results showed that the recombinant protein 3AB was expressed as a formation of inclusion bodies in Escherichia coli. The purified protein could specificially react with FMDV infection antibodies in Western blotting assay, but no reaction with the immune antibodies induced with vaccine. Two assays were no significant differences in specificity and sensitivity for detection of field samples (P>0.05). Therefore, we speculated that the recombinant protein 3AB is a promising molecular marker, which may effectively differentiate FMD-infected from vaccinated animals in a herd. PMID- 21648222 TI - Decisions that put patients at risk. PMID- 21648223 TI - Imatinib and inoperable or metastatic gastrointestinal stromal tumours. Longer follow-up confirms the overall survival benefit. AB - In 2002, patients with inoperable or metastatic gastrointestinal stromal tumours had a median overall survival time of about 19 months with available treatment options. When it was first marketed in this setting in 2002, the efficacy of imatinib, a tyrosine kinase inhibitor, was mainly based on one trial evaluating a surrogate endpoint. This review examines new data, focusing on overall survival. Longer-term data have been published for 56 patients enrolled in a trial comparing imatinib doses of 400 mg and 600 mg per day. The median overall survival time was 57 months and the median progression-free survival time was about 24 months, with no statistically significant difference between the two doses. Two randomised unblinded trials with similar designs compared imatinib doses of 400 mg and 800 mg/day in a total of more than 1600 patients. Combined analysis of the two trials showed no statistically significant difference in median overall survival between the two doses (48.8 months). In contrast, median progression-free survival was significantly longer with the higher dose, by about 4 months. Another trial compared continued treatment versus imatinib withdrawal after a year of treatment in patients whose tumour had stabilised on imatinib 400 mg/day. Imatinib withdrawal was associated with a higher risk of progression. However, when treatment was resumed at a dose of 400 mg/day after disease progression in 26 patients, the median overall survival time was similar to that in patients receiving continuous treatment. Initially, clinical evaluation showed that imatinib provoked many adverse effects, some of which were potentially severe. Cases of heart failure, secondary malignancies, bone remodelling and hepatic and ovarian disorders have since been reported. In the two trials comparing daily imatinib doses of 400 mg and 800 mg, about half of the patients experienced a serious adverse effect. The frequency of serious adverse effects was significantly higher with a daily dose of 800 mg than with 400 mg. The mortality rate was higher with imatinib 800 mg/day. In practice, compared with cytotoxic chemotherapy, imatinib provides a tangible overall survival benefit in patients with inoperable or metastatic gastrointestinal stromal tumours, at a cost of varied, frequent and potentially life-threatening adverse effects. PMID- 21648224 TI - Pazopanib. Kidney cancer: many risks, but is there a benefit for patients? AB - Chemotherapy has little impact on renal carcinoma. Interferon alfa is the standard treatment, in the absence of a better alternative, but the median survival time is less than a year among patients with advanced-stage or metastatic disease. Pazopanib inhibits multiple receptor tyrosine kinases, including the tyrosine kinase coupled to the vascular endothelial growth factor receptor (VEGFR), thus inhibiting angiogenesis. It is authorised in the European Union for the treatment of advanced-stage renal carcinoma. Clinical evaluation of pazopanib is based on a double-blind placebo-controlled trial in 435 patients. Follow-up is currently too short to determine whether pazopanib improves overall survival. The median progression-free survival time, based on radiological criteria, was 5 months longer with pazopanib: 9.2 months versus 4.2 months with placebo. This is similar to the efficacy obtained in trials of other cytotoxic drugs such as sunitinib. The adverse-effect profile of pazopanib is unfavourable: it includes disorders linked to its antiangiogenic activity (hypertension, arterial thrombosis, myocardial infarction, haemorrhage, etc.), as well as gastrointestinal disorders and hand-foot syndrome. In clinical trials including a total of 593 patients, 2 patients receiving pazopanib developed torsades de pointes and 3 other patients died of liver failure. Numerous pharmacokinetic interactions are likely, notably involving cytochrome P450 isoenzyme CYP 3A4. The risk-benefit balance of pazopanib is currently too unfavourable to justify its use in patients with advanced-stage or metastatic kidney cancer. PMID- 21648225 TI - Amlodipine + valsartan + hydrochlorothiazide. A 3-drug fixed-dose combination for hypertension: too many drawbacks. PMID- 21648226 TI - Botulinum toxin type A in children. New indication for limb spasticity. PMID- 21648227 TI - Inhibition of lactation: risks associated with dopamine agonists. PMID- 21648228 TI - Advantages of adverse-effect reporting by patients. PMID- 21648229 TI - Methods for improving the reporting of adverse effects. PMID- 21648230 TI - Preventing neonatal group B streptococcal infection. Intrapartum antibiotic prophylaxis in some high-risk situations. AB - Group B streptococci (GBS) are the leading cause of life-threatening neonatal bacterial infections in developed countries. The newborn is initially colonised during passage through the birth canal. Maternal vaginal carriage is usually asymptomatic. How safe and effective are strategies aimed at preventing severe neonatal GBS infection? To answer these questions, we conducted a review of the literature using the standard Prescrire methodology. In France, Group B streptococci are present in the vagina of about 10-15% of women towards the end of pregnancy. Mother-to-child transmission can occur when the membranes rupture, or during delivery. About one-third of infants born to mothers who carry Group B streptococci are colonised at birth. GBS colonisation rarely affects the newborn's health but, during the first 7 days of life, about 3% of colonised children develop a serious early-onset infection, particularly meningitis, which may be fatal or leave sequelae. Late-onset infections (after 7 days of life) do not appear to be linked to intrapartum colonisation. The risk of early neonatal GBS infection increases in cases of preterm delivery, maternal fever during delivery, and membrane rupture more than 18 hours before delivery. These situations account for 50% to 75% of early neonatal GBS infections. Several randomised trials suggest that intravenous antibiotic prophylaxis in women who carry Group B streptococci, from the onset of labour until delivery, reduces the risk of early-onset neonatal GBS infection from 4.7% to 0.4% (p = 0.02). Other antibiotic strategies are less well assessed or appear to be less effective. Penicillin G (benzylpenicillin) is the antibiotic of choice, while penicillin A (ampicillin or amoxicillin) is an alternative. In case of penicillin allergy, erythromycin or clindamycin are generally active against Group B streptococci and carry no particular risks for the infant. The greatest risk associated with penicillin, especially injectable forms, is an anaphylactic reaction, which can have severe consequences for both mother and child. The estimated frequency is about 5 cases per 10 000 treatments. The possible long-term adverse effects of antibiotic exposure during delivery are poorly documented in children. We found no randomised trials of standard vaginal screening for Group B streptococci between 35 and 38 weeks of gestation, or of a rapid PCR (polymerase chain reaction) test at the time of delivery. In a retrospective study conducted in the US, screening followed by antibiotic therapy if the results were positive was associated with a lower risk of early neonatal infection, but the methodology does not allow firm conclusions to be drawn. The risk-benefit balance has not been determined in terms of neonatal mortality. Widespread implementation of screening guidelines coincided with a decline in the incidence of early neonatal GBS infections in several countries. In the United Kingdom, systematic antibiotic therapy is recommended in high-risk situations, without systematic screening for Group B streptococci. In practice, the first priority is to identify situations in which there is a high risk of neonatal GBS infection, and to administer antibiotics during labour, after screening for GBS carriage, if possible. Outside of these situations, the risk of an anaphylactic reaction must be minimised by choosing the prophylactic antibiotic based on maternal allergy history, and by avoiding antibiotic prophylaxis altogether if the mother has a history of anaphylaxis, whatever the cause. PMID- 21648231 TI - Type 2 diabetes: targeting blood pressure below 140/80 mmHg. PMID- 21648232 TI - Informing patients of the long-term effects of treatments for localised prostate cancer. PMID- 21648233 TI - 1981-2010: 30 years of Prescrire Drug Awards. PMID- 21648234 TI - 2010 Prescrire Drug Awards. PMID- 21648236 TI - 2010 Prescrire Information Awards. PMID- 21648235 TI - 2010 Prescrire Packaging Awards. PMID- 21648237 TI - Spatial and temporal patterns of scleractinian coral, soft coral, and zoanthid disease on a remote, near-pristine coral reef (Palmyra Atoll, central Pacific). AB - There is an urgent need for accurate baselines of coral disease prevalence across our oceans in order for sudden or unnatural changes to be recognized. Palmyra Atoll allows us to study disease dynamics under near-pristine, functionally intact conditions. We examined disease prevalence among all known species of scleractinian coral, soft coral and zoanthid (Palythoa) at a variety of coral reef habitats at Palmyra over a 2 yr period. In 2008, overall disease prevalence across the atoll was low (0.33%), but higher on the shallower backreef (0.88%) and reef terrace (0.80%) than on the deeper forereef (0.09%). Scleractinian coral disease prevalence was higher (0.30%) than were soft coral and zoanthid disease (0.03% combined). Growth anomalies (GAs) were the most commonly encountered lesions, with scleractinian species in the genera Astreopora (2.12%), Acropora (1.30%), and Montipora (0.98%) showing the highest prevalence atoll-wide. Discoloration necrosis (DN) was most prevalent in the zoanthid Palythoa tuberculosa (1.18%), although the soft coral Sinulana and Montipora also had a prevalence of 0.44 and 0.01%, respectively. Overall disease prevalence within permanently marked transects increased from 0.65% in 2008 to 0.79% in 2009. Palythoa DN contributed most to this increased prevalence, which coincided with rising temperatures during the 2009 El Nino. GAs on the majority of susceptible genera at Palmyra increased in number over time, and led to tissue death. Host distribution and environmental factors (e.g., temperature) appear to be important for determining spatiotemporal patterns of disease at Palmyra. More sophisticated analyses are required to tease apart the likely inter-correlated proximate drivers of disease occurrence on remote, near-pristine reefs. PMID- 21648238 TI - Comparison of sensitivity between real-time detection of a TaqMan assay for Batrachochytrium dendrobatidis and conventional detection. AB - A sensitive and quantitative TaqMan assay for the causative agent of chytridiomycosis in amphibians (Batrachochytrium dendrobatidis) has been developed and is routinely used in diagnostic laboratories. We assessed whether the real time detection of the TaqMan assay was as sensitive as the detection of the PCR product by agarose gel electrophoresis and ethidium bromide staining. We found, for practical purposes, that gel-based detection of the diagnostic fragment produced by means of the TaqMan assay or by conventional PCR that used a different polymerase and reaction mix was as sensitive as the real-time detection of the TaqMan assay. We recommend the qualified use of conventional PCR amplification combined with agarose gel electrophoresis and ethidium bromide staining for studies where only prevalence data are required, funding for equipment is limited or the acquisition of a real-time system is not cost effective. PMID- 21648239 TI - Detection of the oyster herpesvirus in commercial bivalve in northern California, USA: conventional and quantitative PCR. AB - The ostreid herpesvirus (OsHV-1) and related oyster herpesviruses (OsHV) are associated with world-wide mortalities of larval and juvenile bivalves. To quantify OsHV viral loads in mollusc tissues, we developed a SYBR Green quantitative PCR (qPCR) based on the A-region of the OsHV-1 genome. Reaction efficiency and precision were demonstrated using a plasmid standard curve. The analytical sensitivity is 1 copy per reaction. We collected Crassostrea gigas, C. sikamea, C. virginica, Ostrea edulis, O. lurida, Mytilus galloprovincialis, and Venerupis phillipinarum from Tomales Bay (TB), and C. gigas from Drakes Estero (DE), California, U.S.A., and initially used conventional PCR (cPCR) to test for presence of OsHV DNA. Subsequently, viral loads were quantified in selected samples of all tested bivalves except O. lurida. Copy numbers were low in each species tested but were significantly greater in C. gigas (p < 0.0001) compared to all other species, suggesting a higher level of infection. OsHV DNA was detected with cPCR and/or qPCR and confirmed by sequencing in C. gigas, C. sikamea, C. virginica, O. edulis, M. galloprovincialis, and V phillipinarum from TB and C. gigas from DE. These data indicate that multiple bivalve species may act as reservoirs for OsHV in TB. A lack of histological abnormalities in potential reservoirs requires alternative methods for their identification. Further investigation is needed to determine the host-parasite relationship for each potential reservoir, including characterization of viral loads and their relationship with infection (via in situ hybridization), assessments of mortality, and host responses. PMID- 21648240 TI - Short-term infection of striped bass Morone saxatilis with Mycobacterium marinum. AB - Striped bass Morone saxatilis were studied in order to characterize their immune responses over the short term following challenge with Mycobacterium marinum. The expression of immunity-related genes (IL-1beta, TNF-alpha, Nramp and TGF-beta) quickly increased following infection with M. marinum, but these genes were subsequently down-regulated despite the fact that bacterial counts remained high. The number of monocytes and neutrophils also initially increased at 1 d postinfection. This confirms the importance of these types of cells in initial inflammation and mycobacterial infection in striped bass. The phagocytic index of splenic leukocytes over these same time frames did not change significantly following infection. The discrete window in which inflammatory mechanisms were stimulated in striped bass may be related to the intracellular nature of this pathogen. PMID- 21648241 TI - Histochemical and ultrastructural analysis of pathology and cell responses in gills of channel catfish affected with proliferative gill disease. AB - Pond-reared channel catfish Ictalurus punctatus with proliferative gill disease (PGD), caused by the myxozoan parasite Henneguya spp., were examined with light and transmission electron microscopy to better characterize the inflammatory response during infection. The early stages of disease are characterized by the destruction of collagen in the matrix of the gill filament cartilage causing weakness and breaks within the gill filaments. These early lesions lacked a notable inflammatory response around the disrupted cartilage, a chondrocyte response was not apparent, and the parasite was not present, suggesting that the cartilage breaks occur prior to inflammation and arrival of the parasite in the gill. In later lesions, a significant inflammatory response was generated in areas of disrupted cartilage, and the inflammatory infiltrate was composed of a mixed population of granulocytes including neutrophils and cells that resembled eosinophils. The majority of eosinophil-like cells demonstrated evidence of degranulation. Trophozoites of Henneguya spp. were surrounded by a uniform population of cells believed to be neutrophils. The granulocytes were infiltrated within the dense collagen layer of the gill filament cartilage and often appeared within chondrocyte lacunae in place of the chondrocyte. The gill lamellae adjacent to the lesions were fused and contained an inflammatory infiltrate containing granulocytes and cells with pericentriolar granules that resembled previous descriptions of Langerhans-like cells. These cells were abundant within damaged lamellar epithelium, but were only rarely found within the gill filament. Lesions that appeared to be recovering lacked the dense collagenous layer around the cartilage and contained hyperplastic and hypertrophic chondrocytes that formed a callus. Other chondrocytes in the lesions had ultrastructural features indicative of cell death. PMID- 21648242 TI - Identification of differentially expressed genes in parasitic phase Miamiensis avidus (Ciliophora: Scuticociliatia) using suppression subtractive hybridization. AB - Miamiensis avidus, a causative agent of scuticociliatosis in cultured marine fish, can live not only in seawater as a free-living organism but also in fish as a parasite. In this study, a cDNA library of representative mRNAs more specific to parasitic phase M. avidus was generated using suppression subtractive hybridization (SSH), and 520 clones selected from the SSH library were single-run sequenced. The differential gene expression patterns were confirmed by semi quantitative reverse-transcription PCR. Of the 510 SSH clones, 21 clones of 6 putative genes did not match sequences in the public database. The expectation values (E-values) of 117 clones encoding 9 putative genes were greater than 1 x 10(-5). The other 372 clones that met the criterion of E value <1 x 10-5 were matched to 26 known sequences in the database. Genes associated with signal transduction, cell proliferation, membrane transportation, protein translocation, and transcription regulation were preferentially expressed in parasitic phase M. avidus. The differential gene expression may be needed for the ciliates to survive in the host fish, and the corresponding proteins might be used as antigen candidates for development of scuticociliatosis vaccines. PMID- 21648243 TI - Histopathological survey of lesions and infections affecting sick ornamental fish in pet shops in New South Wales, Australia. AB - The objective of this study was to describe the frequency of histopathological lesions and categorize histopathologically evident infections in sick ornamental fish from pet shops in New South Wales, Australia. We examined 108 fish that had evidence of morbidity or mortality, including 67 cyprinids, 25 osphronemids, 11 poeciliids, 4 characids and 1 cichlid, sourced from 24 retail outlets. Conditions frequently observed in the study population included branchitis (62/86, 72.1%), visceral granulomas (41/108, 38.0%), dermatitis (17/55, 30.9%), wasting (31/108, 28.7%), and intestinal coccidiosis (18/104, 17.4 %). Branchitis and dermatitis were usually due to monogenean flukes, or flagellate or ciliate protozoa. Intralesional Microsporidia (16/41, 39.0%), mycobacteria (7/41, 17.%), or Myxosporidia (5/41, 12.2%) were identified in the majority of fish with visceral granulomas; however, special stains were critical in their identification. The proportion of histologically evident infections was remarkably high (77/108, 71.3%), and parasitic infections predominated. Many pathogens identified in the study have low host specificity and/or direct life cycles which would facilitate transmission to exposed naive fish populations, potentially posing a threat to native and commercial fish populations. Those caring for sick ornamental fish should take appropriate steps to investigate infectious disease and should take precautions that prevent the spread of pathogens. PMID- 21648244 TI - Review of Panulirus argus virus 1--a decade after its discovery. AB - In 2000, a pathogenic virus was discovered in juvenile Caribbean spiny lobsters Panulirus argus from the Florida Keys, U.S.A. Panulirus argus virus 1 (PaV1) is the first naturally occurring pathogenic virus reported from lobsters, and it profoundly affects their ecology and physiology. PaV1 is widespread in the Caribbean with infections reported in Florida (U.S.A.), St. Croix, St. Kitts, Yucatan (Mexico), Belize, and Cuba. It is most prevalent and nearly always lethal in the smallest juvenile lobsters, but this declines with increasing lobster size; adults harbor the virus, but do not present the characteristic signs of the disease. No other PaV1 hosts are known. The prevalence of PaV1 in juvenile lobsters from the Florida Keys has been stable since 1999, but has risen to nearly 11% in the eastern Yucatan since 2001. Heavily infected lobsters become sedentary, cease feeding, and die of metabolic exhaustion. Experimental routes of viral transmission include ingestion, contact, and for newly settled juveniles, free virus particles in seawater. Prior to infectiousness, healthy lobsters tend to avoid diseased lobsters and so infected juvenile lobsters mostly dwell alone, which appears to reduce disease transmission. However, avoidance of diseased individuals may result in increased shelter competition between healthy and diseased lobsters, and greater predation on infected lobsters. Little is known about PaV1 outside of Mexico and the USA, but it poses a potential threat to P. argus fisheries throughout the Caribbean. PMID- 21648245 TI - Microsatellite marker development in the protozoan parasite Perkinsus olseni. AB - The analysis of an enriched partial genomic library and of public expressed sequence tag (EST) resources allowed the characterization of the first microsatellite loci in the protozoan parasite Perkinsus olseni. Clonal cultures from laboratory isolates derived from infected clams Ruditapes decussatus (from Spain), R. philippinarum (from Spain and Japan), and Austrovenus stutchburyi (from New Zealand) were used for the characterization of 12 microsatellites. Low variation was detected at most loci, with the number of alleles at polymorphic loci ranging from 2 to 7 (average 3.20 +/- 0.51) and gene diversity from 0.11 to 0.79 (average 0.40 +/- 0.07). Preliminary results show that (1) isolates of P. olseni are diploid cells, and (2) multiple infections can occur within a single host. Eight of the loci analyzed successfully cross-amplified in the congeneric species P. mediterraneus. These microsatellite markers will be useful to analyze in detail the population genetic structure of P. olseni, crucial for the efficient management of this parasitic disease. PMID- 21648246 TI - In vitro treatments for the theront stage of the ciliate protozoan Cryptocaryon irritans. AB - The ciliate protozoan Cryptocaryon irritans Brown, 1951, the 'marine white spot', causes one of the most important parasitic fish diseases, with extensive losses every year in mariculture and in the ornamental fish industry. In the present study, we explore the in vitro use of 8 different compounds against the theront (infective) stage of C. irritans; these compounds include extracts of natural products (epigallocatechin gallate (EGCG), L-DOPA, papain), peracetic acid-based compounds (Proxitane 5:23 and 15% peracetic acid, PAA), quinine-based compounds (quinacrine hydrochloride and chloroquine diphosphate) and hydrogen peroxide. All of these compounds had an effect on theront survival; however, only EGCG caused significant theront mortality when applied in doses > or =50 mg l(-1) and over a period of 3 h; papain caused a maximum theront mortality of <50%. We discuss the type of application and potential utility of the compounds tested as part of a management control strategy for C. irritans infections in marine aquaculture and the ornamental fish industry. PMID- 21648247 TI - Children of misfortune: early adversity and cumulative inequality in perceived life trajectories. AB - Adversity early in life may alter pathways of aging, but what interpretive processes can soften the blow of early insults? Drawing from cumulative inequality theory, the authors analyze trajectories of life evaluations and then consider whether early adversity offsets favorable expectations for the future. Results reveal that early adversity contributes to more negative views of the past but rising expectations for the future. Early adversity also has enduring effects on life evaluations, offsetting the influence of buoyant expectations. The findings draw attention to the limits of human agency under the constraints of early adversity-a process described as biographical structuration. PMID- 21648248 TI - Income inequality and income segregation. AB - This article investigates how the growth in income inequality from 1970 to 2000 affected patterns of income segregation along three dimensions: the spatial segregation of poverty and affluence, race-specific patterns of income segregation, and the geographic scale of income segregation. The evidence reveals a robust relationship between income inequality and income segregation, an effect that is larger for black families than for white families. In addition, income inequality affects income segregation primarily through its effect on the large scale spatial segregation of affluence rather than by affecting the spatial segregation of poverty or by altering small-scale patterns of income segregation. PMID- 21648249 TI - Moving teenagers out of high-risk neighborhoods: how girls fare better than boys. AB - Moving to Opportunity (MTO) offered public housing residents the opportunity to move to low-poverty neighborhoods. Several years later, boys in the experimental group fared no better on measures of risk behavior than their control group counterparts, whereas girls in the experimental group engaged in lower-risk behavior than control group girls. The authors explore these differences by analyzing data from in-depth interviews conducted with 86 teens in Baltimore and Chicago. They find that daily routines, fitting in with neighborhood norms, neighborhood navigation strategies, interactions with peers, friendship making, and distance from father figures may contribute to how girls who moved via MTO benefited more than boys. PMID- 21648250 TI - Cultural mechanisms and the persistence of neighborhood violence. AB - Sociologists have given considerable attention to identifying the neighborhood level social-interactional mechanisms that influence outcomes such as crime, educational attainment, and health. Yet, cultural mechanisms are often overlooked in quantitative studies of neighborhood effects. This paper adds a cultural dimension to neighborhood effects research by exploring the consequences of legal cynicism. Legal cynicism refers to a cultural frame in which people perceive the law as illegitimate, unresponsive, and ill equipped to ensure public safety. The authors find that legal cynicism explains why homicide persisted in certain Chicago neighborhoods during the 1990s despite declines in poverty and declines in violence city-wide. PMID- 21648251 TI - Segregation in social networks based on acquaintanceship and trust. AB - Using 2006 General Social Survey data, the authors compare levels of segregation by race and along other dimensions of potential social cleavage in the contemporary United States. Americans are not as isolated as the most extreme recent estimates suggest. However, hopes that "bridging" social capital is more common in broader acquaintanceship networks than in core networks are not supported. Instead, the entire acquaintanceship network is perceived by Americans to be about as segregated as the much smaller network of close ties. People do not always know the religiosity, political ideology, family behaviors, or socioeconomic status of their acquaintances, but perceived social divisions on these dimensions are high, sometimes rivaling racial segregation in acquaintanceship networks. The major challenge to social integration today comes from the tendency of many Americans to isolate themselves from others who differ on race, political ideology, level of religiosity, and other salient aspects of social identity. PMID- 21648252 TI - Protestant responses to Darwinism in Denmark, 1859-1914. PMID- 21648253 TI - Transformation of a profession: an overview of the 20th century. PMID- 21648254 TI - Lost in the patent office--a whimsical journey through pharmaceutical patents. Episode two: poison bottle stoppers. PMID- 21648255 TI - Our future starts here. PMID- 21648256 TI - Pediatrics looks ahead to the medical home. PMID- 21648257 TI - Your business is our business: pediatrics, public health. PMID- 21648258 TI - The sisters knew: a child needs a home. PMID- 21648259 TI - Mercy adapts services to region's needs. PMID- 21648260 TI - Parents and young patients as partners in care. PMID- 21648261 TI - Hospital Care--a child's-eye view. PMID- 21648262 TI - Coordinated perinatal care pays off for mother and child. PMID- 21648263 TI - Program puts 'physical' back in education. PMID- 21648264 TI - Childhood obesity is a family fight--one hospital's way. PMID- 21648265 TI - Pediatric safety focus earns kudos. PMID- 21648266 TI - Beyond borders: pediatric care goes overseas. PMID- 21648267 TI - Team-teaching about palliative care for kids. PMID- 21648268 TI - Child trafficking: what Catholic health care needs to know. PMID- 21648269 TI - Religion, spirituality and the search for common ground. PMID- 21648270 TI - IRS Form 990, Schedule H: address these questions up front. PMID- 21648271 TI - IRS Form 990, Schedule H: a deep dive: ready, set, go. PMID- 21648272 TI - Beyond the IRS. How does your public report stack up? PMID- 21648273 TI - Breaking bonds at what expense? PMID- 21648274 TI - At mother's bedside, lessons hit home. PMID- 21648275 TI - What brings a child to the doctor? A lesson from hyperthyroidism. PMID- 21648276 TI - Hypothalamic hamartoma causing precocious puberty in a female child. PMID- 21648277 TI - Adiponectin moderates the relationship between adiposity and leptin in adolescents regardless of gender or race. AB - OBJECTIVES: To determine gender or race differences in associations between adiposity and leptin, and whether adiponectin moderates these relationships. METHODS: Subjects were 441 adolescents, 14-18 years old (44% black, 56% white; 50% female, 50% male). Percent body fat (%BF) was measured with dual-energy X-ray absorptiometry. Leptin and adiponectin were measured using immunoassays. RESULTS: Among the four groups (white boys, white girls, black boys and black girls), white girls had the highest adiponectin (p = 0.0017) and black girls had the highest leptin (p = 0.0164). Percent BF and leptin were positively correlated (p = 0.0164). The %BF-leptin relationship was stronger in boys than girls (p < 0.0001). Those with lower adiponectin had a stronger %BF-leptin relationship than those with high adiponectin in the entire sample (p = 0.0220). Statistical models were adjusted for age, race, gender and the interaction between race and gender. CONCLUSION: Our data suggest a protective metabolic interaction for adiponectin and lend additional support for obesity prevention strategies in adolescents. PMID- 21648278 TI - Obesity in children and adolescents: relationship to growth, pubarche, menarche, and voice break. AB - OBJECTIVE: The relationships between obesity, pubertal development, and height are controversial. Therefore, we compared the prevalence of pubarche, menarche, and voice break between a large collective of obese and normal-weight children and adolescents aged 10-16 years. METHODS: We assessed weight, height, pubarche, menarche, and voice break in 1383 obese German children and in 6615 children of a representative national German cohort aged 10-16 years. In all obese children, gonadotropins were determined and birth weight data were collected. RESULTS: Independently of gender, the height standard deviation score (SDS) was significantly greater (0.3-1.0) in obese children <14 years compared to the reference cohort. Independently of age, the percentage of obese boys with pubarche was significantly lower compared to age-matched normal-weight boys. In girls <13 years, the prevalence of obese girls with pubarche was significantly lower compared to age-matched normal-weight girls. In boys > or =11 years, the percentage of obese boys with change of voice was significantly lower compared to age-matched normal-weight boys. In girls > or =11 years, the prevalence of obese girls with menarche was significantly lower compared to age-matched normal-weight girls. Birth weight had no impact on pubarche in the obese children. Luteinizing hormone was > 0.3 IU/L in 86% of the children > or =10 years with pubarche. CONCLUSIONS: Obese children are taller than normal-weight children up to the age of 14 years. Since obese children demonstrated pubarche, menarche, and voice break later than their normal-weight peers, the increase in height in obese children does not seem to be attributable to earlier onset of puberty. PMID- 21648279 TI - Studies of different female rat models of hypothalamic obesity. AB - Hypothalamic obesity (HO) is a major and unsolved problem in patients with medial hypothalamic lesions and is associated with hyperinsulinemia and hyperleptinemia. The purpose of this study was to create a rodent model that mimics metabolic changes in HO for use in therapeutic testing. Female Sprague-Dawley rats were used to test the individual and combined effects of two types of medial hypothalamic lesions: arcuate nucleus (ARC) lesions by injection of monosodium glutamate at neonatal age, and ventromedial nucleus (VMN) lesions by passing an anodal current through an electrode placed in the VMN at age 80 days. Adiposity in ARC-lesioned animals was associated with decreased food intake and stunted growth, while VMN lesions were associated with hyperphagia but not reduced growth. The greatest weight gain (weight at age 200 days 712 +/- 65 vs. 451 +/- 19 g in controls), hyperphagia (food intake 10 days following surgery 33 +/- 0.8 vs. 18.5 +/- 0.7 g/day in sham-treated rats), hyperinsulinemia and hyperleptinemia occurred in rats that received both ARC and VMN lesions. Thus, the combined medial hypothalamic lesions result in an obesity phenotype similar to that of patients that suffer from HO and are consequently more suitable for testing potential therapeutics for this disorder than lesions of single hypothalamic nuclei. PMID- 21648280 TI - Frequency of congenital hypothyroidism in neonates in the Konya region, Turkey. AB - Congenital hypothyroidism (CH) resulting from deficient production of thyroid hormone is one of the most commonly encountered diseases in pediatric endocrinology. Thyroid hormones play a crucial role in normal cerebral and growth maturation. These harmful effects on the cerebral and growth maturation can be prevented by early diagnosis and sufficient treatment in the first weeks of life. Diagnosis must be determined immediately within days after birth and effective treatment must begin. Unfortunately, despite the presence of national neonatal screening programs, CH cases are still rarely seen. In our study, it was aimed to assess the outcome of having determined an early diagnosis of CH and initiating treatment with thyroid stimulating hormone (TSH) screening test on live born babies over a period of 7 years in our hospital. With this aim, 93,897 live births were evaluated in the Doctor Faruk Sukan Obstetrics and Pediatrics Hospital between the years of 1999 and 2007. All neonates were screened with the TSH test. CH was determined in 43 (1/2183) of all cases and treatment was begun. The importance of this test was emphasized in that the test should be performed routinely on all neonates to obtain an early diagnosis and so that treatment for CH can begin. PMID- 21648281 TI - Thyroid dysfunctions in children detected in mass screening for congenital hypothyroidism. AB - BACKGROUND: Congenital hypothyroidism (CH) affects approximately 1:3000-1:4000 infants. OBJECTIVES: To determine the prevalence of CH and isolated hyperthyrotropinemia (IHT) in newborns selected in mass screening for CH. METHODS: Mass screening of 233,120 neonates born in southeastern Poland was carried out and CH-suspected children were identified. Serum thyroid-stimulating hormone and free thyroxine levels were determined during first confirmation and diagnosis re-evaluation in 118 and 34 children, respectively. Additionally, the patients were subjected to thyroid ultrasonography (n=53) and/or scintiscan (n=28). RESULTS: Out of 118 children, first confirmation indicated CH in 58 neonates and IHT in 4 neonates. Out of these, 34 were re-evaluated with regard to diagnosis. A final diagnosis of permanent CH was reported in 34 children with thyroid dysgenesis (n=27) or dyshormonogenesis (n=7), transient CH affected 15 children, and permanent IHT was diagnosed in 6 children. CH prevalence was 1:4570 (permanent 1:6475, transient 1:38,853) and permanent IHT 1:38,853. CONCLUSIONS: The prevalence of CH and IHT corresponds to the prevalence of the condition in iodine-sufficient and borderline iodine-deficient areas. PMID- 21648282 TI - A nutritional intervention study with hydrolyzed collagen in pre-pubertal spanish children: influence on bone modeling biomarkers. AB - AIM: The aim of the study was to investigate the influence of dietary intake of commercial hydrolyzed collagen (Gelatine Royal) on bone remodeling in pre pubertal children. METHODS: A randomized double-blind study was carried out in 60 children (9.42 +/- 1.31 years) divided into three groups according to the amount of partially hydrolyzed collagen taken daily for 4 months: placebo (G-I, n=18), collagen (G-II, n=20) and collagen+calcium (G-III, n=22) groups. Analyses of the following biochemical markers were carried out: total and bone alkaline phosphatase (tALP and bALP), osteocalcin, tartrate-resistant acid phosphatase (TRAP), type I collagen carboxy-terminal telopeptide, lipids, calcium, 25 hydroxyvitamin D, insulin-like growth factor-1 (IGF-1), thyroid-stimulating hormone, free thyroxin and intact parathormone. RESULTS: There was a significantly greater increase in serum IGF-1 in G-III than in G-II (p < 0.01) or G-I (p < 0.05) during the study period, and a significantly greater increase in plasma tALP in G-III than in G-I (p < 0.05). Serum bALP behavior significantly (p < 0.05) differed between G-II (increase) and G-I (decrease). Plasma TRAP behavior significantly differed between G-II and G-I (p < 0.01) and between G-III and G-II (p < 0.05). CONCLUSION: Daily dietary intake of hydrolyzed collagen seems to have a potential role in enhancing bone remodeling at key stages of growth and development. PMID- 21648283 TI - Neonatal screening for congenital adrenal hyperplasia: transitory elevation of 17 hydroxyprogesterone. AB - AIM: The aim of the study was to identify patients with transitory elevation (TE) of 17-hydroxyprogesterone (17-OHP) using neonatal screening for congenital adrenal hyperplasia due to 21-hydroxylase deficiency (21-OHD) and to compare them with patients with 21-OHD. METHODS: This was a retrospective study of patients with high 17-OHP levels detected during newborn screening in Madrid, Spain. RESULTS: 17-OHP levels were significantly higher in the 33 21-OHD patients, who tended to present hyponatraemia and hyperkalemia. The TE-17-OHP group was characterized by normal initial physical examination (88.8% vs. 39.4%), lower gestational age and a higher number of stressful perinatal factors. 17-OHP levels decreased spontaneously in this group. Molecular diagnosis allowed us to discard the most frequent mutations associated with 21-OHD. CONCLUSIONS: Newborns with slightly increased 17-OHP levels and normal results for physical examination, acid-base equilibrium, glycemia, electrolytes and perinatal stress factors should be carefully evaluated. Decisions on treatment should be postponed until these results are available. PMID- 21648284 TI - Evidence of insulin-like growth factor binding protein-3 proteolysis during growth hormone stimulation testing. AB - OBJECTIVES: The ternary complex is composed of insulin-like growth factor (IGF) I, IGF binding protein (IGFBP)-3 and acid labile subunit (ALS). Growth hormone (GH) promotes IGFBP-3 proteolysis to release free IGF-I, ALS, and IGFBP-3 fragments. Our aim was to determine whether elevated GH levels during GH stimulation testing would trigger IGFBP-3 proteolysis. DESIGN: This prospective study of 10 short prepubertal children (height standard deviation score -2.37 +/- 0.31) used arginine and GH releasing hormone stimulation to study dynamic changes in the ternary complex moieties. IGFBP-3 was measured in two assays: a radioimmunoassay (RIA) that detects both cleaved and intact IGFBP-3; and an immunochemiluminescence assay (ICMA) that detects only intact IGFBP-3. RESULTS: IGFBP-3 measured by RIA increased by 19% (p < 0.05), while IGFBP-3 measured by ICMA did not significantly increase (6.1%). CONCLUSION: The significant increase in IGFBP-3 measured by RIA, but not ICMA, provides evidence of IGFBP-3 proteolysis during acute GH stimulation. PMID- 21648285 TI - Pediatric 25-hydroxyvitamin D concentrations in neurofibromatosis type 1. AB - OBJECTIVE: Low 25-hydroxyvitamin D (25OHD) concentrations have been associated with tumors and osteopenia or fractures in adults with neurofibromatosis type 1 (NF1). We report 25OHD concentrations in 109 children with NF1 and 218 controls matched for age, sex, geographic location, and time of year. METHODS: Children with NF1 were recruited (n=109; 2-17 years), and clinical data and dual-energy X ray absorptiometry measurements were obtained. 25OHD concentrations were measured in subjects and controls. RESULTS: More NF1 individuals (50%) were in the 25OHD insufficient or deficient range (<30 ng/mL) (1 ng/mL = 2.496 nmol/L) compared to controls (36%) (p = 0.0129). 25OHD concentrations were higher in individuals with neurofibromas after controlling for age (p = 0.0393), and were negatively associated with whole-body subtotal bone mineral density (BMD) z-scores (p = 0.0385). CONCLUSIONS: More children with NF1 had 25OHD concentrations <30 ng/mL, potentially because of increased pigmentation and/or decreased sunlight exposure. In contrast to adults, decreased 25OHD concentrations were not associated with neurofibromas, and there was no positive association between 25OHD and BMD. PMID- 21648286 TI - Temperature-responsive and biodegradable PVA:PVP k30:poloxamer 407 hydrogel for controlled delivery of human growth hormone (hGH). AB - Recombinant human growth hormone (rhGH) is used for replacement therapy of pediatric hypopituitary dwarfism. Growth rate in children was observed to be better on the daily injection schedule compared with the currently used therapeutic regimen of thrice a week injection. Thus, a controlled release formulation would overcome the drawback of traditional rhGH therapy such as the need for multiple injections. Poloxamers are a family of triblock copolymers consisting of two hydrophilic blocks of polyoxyethylene separated by a hydrophobic block of polyoxypropylene, which form micelles at low concentrations and form clear thermally reversible gels at high concentrations. We used poloxamer gels to develop a controlled release formulation of hGH. The objective of this study was to develop an in situ gel forming drug delivery system for hGH using the minimum possible ratio of poloxamer 407 (P407). Decreasing the concentration of poloxamer could reduce the risk of hypertriglyceridemia induction. Different additives were added to the poloxamer formulations. It was observed that among different additives polyvinylpyrrolidone k30 (PVP k30) and polyvinyl alcohol (PVA) decrease poloxamer concentration required to form in situ gelation from 18% to 10%. The dynamic viscoelastic properties of the samples were determined. Both the storage modulus and the loss modulus of the samples increased abruptly as the temperature passed a certain point. It can be concluded that combining P407 and PVP and PVA could be a promising strategy for preparation of thermally reversible in situ gel forming delivery systems of hGH with low poloxamer concentration. PMID- 21648287 TI - A novel EIF2AK3 mutation leading to Wolcott-Rallison syndrome in a Chinese child. AB - OBJECTIVE: Mutations in the EIF2AK3 gene are known to cause Wolcott-Rallison syndrome (WRS), a rare recessive disorder characterized by early-onset diabetes, skeletal abnormalities, and liver dysfunction. To date, studies on WRS have revealed several mutation types leading to onset of the disease. In the present study, we analyzed the EIF2AK3 gene in a 10-year-old WRS patient and his parents to study the clinical features and the mechanism for genetic onset of WRS. METHOD: A patient diagnosed with WRS and his parents were chosen as research subjects. PCR techniques were used to amplify the 17 exons of the EIF2AK3 gene and DNA direct assay techniques were used for gene mutation analysis. RESULT: Gene mutation analysis revealed a 1798 A/T heterozygous mutation in exon 9 of the patient's EIF2AK3 gene. This nonsense mutation can lead to a C-stop and result in a truncated protein of 532 amino acid residues in length (C532STOP). The patient's parents are nonconsanguineous and the patient's father carries the same mutation, while the mother carries no EIF2AK3 mutation. CONCLUSION: ETF2AK3 gene mutations can lead to the onset of WRS. The study results provide knowledge that furthers our understanding of the genetic mechanism of WRS. PMID- 21648288 TI - Neurological Wilson's disease with refractory rickets. AB - Wilson's disease rarely presents with isolated neurological complaints without any hepatic involvement. Refractory rickets with Wilson's disease has been infrequently reported in literature. We are reporting a case of isolated neurological Wilson's disease associated with refractory rickets which on complete evaluation was diagnosed as familial hypophosphatemic rickets. PMID- 21648289 TI - HNF1A mutation presenting with fetal macrosomia and hypoglycemia in childhood prior to onset of overt diabetes. AB - BACKGROUND: HNF1A-MODY (MODY3) is a common subtype of autosomal dominant diabetes. Unlike HNF4-MODY where fetal macrosomia and early postnatal hyperinsulinemic hypoglycemia have been reported, history of transient insulin overproduction has not yet been recognized in individuals with HNF1A-MODY. CASE REPORT: Here, we report on a 40-year-old male patient with HNF1A mutation p.Arg272His (c.815G>A) having a history of fetal macrosomia (4750 g, 59 cm), and, at least, one attack of symptomatic hypoglycemia in childhood. Diabetes was subsequently diagnosed at 19 years of age. The proband's daughter who developed diabetes at 16 years carries the same mutation, but her birth weight and length were in the upper normal range, and she never experienced hypoglycemic symptoms. CONCLUSION: The observation of fetal macrosomia and hypoglycemia in childhood is indicative of a biphasic impact of the HNF1A mutation on p-cell function over the lifespan, leading from inappropriate insulin oversecretion to final clinical diabetes. PMID- 21648290 TI - Pediatric Cushing's disease due to pituitary hyperplasia. PMID- 21648291 TI - A 12-year-old chinese girl with Cushing syndrome and virilization due to adrenocortical carcinoma. AB - A Chinese adolescent girl presented with secondary amenorrhea. During follow-up, she gradually developed Cushingoid features and virilization. After a series of endocrine investigations, including urinary steroid profiling, a diagnosis of adrenocortical carcinoma was made. The treatment and prognosis of the disease are discussed. PMID- 21648292 TI - Primary pigmented nodular adrenocortical disease: a case report in a 7-year-old girl. AB - Primary pigmented nodular adrenocortical disease (PPNAD) is a rare cause of Cushing syndrome in children, often occurring in association with Carney complex. We report a case of Cushing syndrome due to isolated non-familial PPNAD. The child presented with typical clinical characteristics, growth retardation and obesity. Liddle's test was positive but micronodular appearance was not evident on CT scan and MRI; selective venous sampling revealed higher cortisol concentrations in the right adrenal vein. The patient underwent a laparoscopic right adrenalectomy. Postoperatively, hypercortisolism signs disappeared but after the second year a slight increase in urinary cortisol was noted and the patient developed osteopenia. Although significant catch-up growth occurred postoperatively, height did not normalize over the next 2 years. When she entered puberty, treatment with a luteinizing-hormone-releasing hormone agonist was initiated and growth hormone was added. Almost 5 years later a left adrenalectomy was also performed. Thereafter, complete disease remission was observed, the patient's growth accelerated and her osteopenia reversed. PMID- 21648293 TI - Normal adrenal function in an infant following a pregnancy complicated by maternal adrenal cortical carcinoma and mitotane exposure. AB - Maternal adrenal cortical carcinoma in pregnancy is rare. We report a case of an infant born to a mother with a history of adrenal cortical carcinoma. The pregnancy was complicated by fetal exposure to mitotane and dexamethasone. Despite the potential teratogenic exposures, there was no evidence of adrenal dysfunction in the infant. Growth and development at 12 months of age are normal and prognosis appears favorable. The long-term impact of fetal exposure to mitotane and glucocorticoid requires further investigation. PMID- 21648294 TI - Transient hyperthyroidism due to acute suppurative thyroiditis in an adolescent female. PMID- 21648295 TI - High frequency of cardiac and behavioral complaints as presenting symptoms of hyperthyroidism in children. AB - OBJECTIVE: Descriptive data characterizing the frequency of various chief complaints and presenting symptomatology in children with hyperthyroidism are lacking. Furthermore, difficulties in recognizing atypical presentations may delay diagnosis and increase morbidity. METHODS: We performed a retrospective review of the medical records of 76 children with hyperthyroidism to characterize their chief complaints at initial presentation to care and document other presenting symptomatology. RESULTS: Cardiac symptoms were the most frequent chief complaint, accounting for 23% of presenting complaints. Major mood and behavior disturbances were also frequently present (21%), but were infrequently cited as the chief complaint (6%). CONCLUSIONS: This study is the first to describe chief complaints separately from the other signs and symptoms of hyperthyroidism noted at the time of presentation to medical attention. Cardiac complaints were the most common: however, complaints associated with behavioral and mood disorders also occurred frequently. Clinicians should be aware of these presentations so that hyperthyroidism is diagnosed promptly to avoid morbidity associated with delayed diagnosis. PMID- 21648296 TI - Aplasia cutis congenita in surviving co-twin after propylthiouracil exposure in utero. AB - AIM: Aplasia cutis congenita (ACC) has been observed after fetal exposure to the antithyroid drug methimazole (MMI), but not reported after propylthiouracil (PTU), the current antithyroid drug of choice during pregnancy. This occurrence has implications for patient information and causal research. CASE REPORT: We describe a surviving term co-twin to a mother with hyperthyroidism exposed to PTU from conception to 34 weeks of gestation presenting with ACC at birth. DISCUSSION: The association between PTU exposure and ACC is clinically relevant and allows speculation on the etiology. A similar mechanism to the classical MMI induced ACC is postulated, unless a vascular etiology suggested by a vanishing twin or maternal hyperthyroidism itself is causal. Coincidence of PTU exposure and ACC seems unlikely. CONCLUSION: ACC in a newborn after PTU exposure during pregnancy hitherto observed only after MMI strongly encourages further reports of similar cases that may remain clinically underdiagnosed or unreported. Such confirmation could have significant implications for maternal treatment of hyperthyroidism, common in women of childbearing age. PMID- 21648297 TI - Cushing syndrome due to ectopic adrenocorticotropic hormone secretion in a 3-year old child. AB - Ectopic adrenocorticotrophic hormone (ACTH) secretion is a rare cause of Cushing syndrome in paediatric age, due to tumours arising from different tissues. To date, only 11 reports of ACTH-secreting pancreatic tumours in children and adolescents exist in the literature. We present a paediatric case of Cushing syndrome caused by ectopic ACTH secretion. This was caused by a large acinar cell carcinoma that developed in the pancreas of a 3-year-old girl. PMID- 21648298 TI - Familial occurrence of Turner syndrome: casual event or increased risk? AB - AIM: The prevalence of Turner syndrome (TS) at birth has been estimated as approximately 1 in 2500 live female births. An increased risk of TS in subsequent pregnancies for couples who already have a daughter with TS has not been reported. METHODS: We reviewed the records of 140 patients to evaluate the presence of familial TS occurrence. RESULTS: Recurrence of TS was observed in 1.4% of our case series, which represents a 35-fold increased probability of having a second child with TS compared to no recurrence. CONCLUSION: This observation suggests that a risk of recurrence is possible, even though it is generally assumed that the likelihood of recurrent pregnancies with TS is similar to that in the general population. A wider study would be useful to confirm these data to improve genetic counseling for families with a daughter with TS. PMID- 21648299 TI - Mature teratoma, a rare cause of virilization in adolescence. AB - Virilization in an adolescent patient can occur for multiple reasons (ovarian, suprarenal or exogenous reasons). We describe a 14-year-old patient with 1-year secondary amenorrhea, who had an ovarian mature teratoma as a cause of her clinical history. PMID- 21648300 TI - Co-existence of ADHD, autoimmune hypothyroidism and pituitary macroadenoma presenting in a behaviour clinic: a case report and brief review of the literature. PMID- 21648301 TI - Virilizing para-adrenocortical adenoma associated with idiopathic-acquired generalized anhidrosis in an adolescent girl. AB - Adrenocortical tumors are rare in childhood and adolescence. Virilization, alone or in combination with signs of overproduction of other adrenal hormones, is the most common clinical presentation. Here we report an unusual case of an African American female adolescent presenting with idiopathic acquired generalized anhidrosis, dysregulation of body temperature, absence of adult body odor and dry skin in the face of a virilizing para-adrenocortical adenoma. Virilization signs regressed soon after removal of the tumor, but normalization of the 3alpha androstenediol glucuronide (3alpha-AG) took longer compared to other measurable androgens; accompanied by anhidrosis. The association of remitting anhidrosis with normalized levels of 3alpha-AG suggests it might be a possible mechanism for anhidrosis. High 3alpha-AG levels might implicate the increased peripheral conversion of weak pro-androgens with different biochemical structure. We recommend obtaining 3alpha-AG beside other androgens in virilized patients with atypical dermatological symptoms in the face of hyperandrogenism. PMID- 21648302 TI - Concurrent occurrence of chronic lymphocytic thyroiditis with hypothyroidism and growth hormone deficiency in a Turner's syndrome patient. AB - BACKGROUND: Concurrent deficiencies of three hormones in patients with Turner's syndrome (TS) have rarely been reported. Here, we describe a case of a young girl who had Turner's syndrome with concomitant chronic lymphocytic thyroiditis, growth hormone deficiency, and hypothyroidism with cardiopericarditis. CASE: An 11-year-old girl was referred to the outpatient clinic because of short stature, ochriasis, and cardiopalmus. Her ultrasound revealed absence of ovarian tissue. Karyotype examination suggested Turner's syndrome with sex hormone deficiency. She was found to have an abnormal thyroid gland and elevated thyroid stimulating hormone (TSH). A positive thyroid autoantibody titer confirmed the diagnosis of chronic lymphocytic thyroiditis with hypothyroidism. Furthermore, her growth hormone levels were well below normal. DIAGNOSIS: A multi-endocrine disorder, i.e., Turner's syndrome with chronic lymphocytic thyroiditis, growth hormone deficiency, and hypothyroidism with cardiopericarditis was diagnosed. Growth hormone and thyroxin substitution therapy was suggested. PMID- 21648303 TI - Patient report: sacral agenesis with hypopituitarism. AB - We report an association of sacral agenesis and hypopituitarism in a child born of a diabetic mother. The child presented with short stature and evaluation revealed sacral agenesis, growth hormone deficiency (GHD) and adrenal insufficiency. This association might be important as short stature in children with sacral agenesis is usually attributed to mechanical factors. Early detection of GHD and treatment might help the child in gaining height. PMID- 21648304 TI - Stimulation of functional vision in children with perinatal brain damage. AB - Cerebral visual impairment (CVI) is one of the most common causes of bilateral visual loss, which frequently occurs due to perinatal brain injury. Vision in early life has great impact on acquisition of basic comprehensions which are fundamental for further development. Therefore, early detection of visual problems and early intervention is necessary. The aim of the present study is to determine specific visual functioning of children with perinatal brain damage and the influence of visual stimulation on development of functional vision at early age of life. We initially assessed 30 children with perinatal brain damage up to 3 years of age, who were reffered to our pediatric low vision cabinet in "Little house" from child neurologists, ophthalmologists Type and degree of visual impairment was determined according to functional vision assessment of each child. On the bases of those assessments different kind of visual stimulations were carried out with children who have been identified to have a certain visual impairment. Through visual stimulation program some of the children were stimulated with light stimulus, some with different materials under the ultraviolet (UV) light, and some with bright color and high contrast materials. Children were also involved in program of early stimulation of overall sensory motor development. Goals and methods of therapy were determined individually, based on observation of child's possibilities and need. After one year of program, reassessment was done. Results for visual functions and functional vision were compared to evaluate the improvement of the vision development. These results have shown that there was significant improvement in functional vision, especially in visual attention and visual communication. PMID- 21648305 TI - Arylsulfatase a gene polymorphisms in relapsing remitting multiple sclerosis: genotype-phenotype correlation and estimation of disease progression. AB - Arylsulfatase A (ASA) is a lysosomal enzyme involved in catabolism of cerebroside sulfate, major lipid constituent of oligodendrocyte membranes. Various polymorphisms in ASA gene have been described, leading to different levels of enzyme deficiency. Progressive demyelination occurs in metachromatic leukodystrophy (MLD), while the condition of ASA-pseudodeficiency (ASA-PD) is suggested to contribute to complex pathogenesis of multiple sclerosis (MS). This work presents usefulness of genotype-phenotype correlation in estimation of disease severity and progression. The presence of two most common mutations associated with ASA-PD was analyzed in 56 patients with diagnosis of relapsing remitting multiple sclerosis, by polymerase chain reaction restriction fragment length polymorphism method. In MS patients confirmed as ASA-PD mutations carriers, arylsulfatase activity was determined in leukocyte homogenates by spectrophotometry. To determine whether there is a difference between disability level and/or disease progression in patients with or without mutations we have estimated disability level using Expanded disability status scale (EDSS) and disease progression using Multiple sclerosis severity score (MSSS). Correlation of genotypes and disease progression was statistically analyzed by Kruskal-Wallis test. Patients showing higher MSSS score and found to be carriers of both analyzed ASA-PD mutations were additionally examined using conventional magnetic resonance (MR) techniques. The presence of either one or both mutations was determined in 13 patients. Lower ASA activities were observed in all MS patients carrying the mutations. Nine of the mutations carriers had mild disability (EDSS=0-4.0), 1 had moderate disability (EDSS=4.5-5.5), and 3 had severe disability (EDSS > or = 6.0). On the other hand, only 3 MS patients who were mutation carriers showed MSSS values lower than 5.000 while in other MS patients mutation carriers the MSSS values ranged from 5.267 to 9.453. Comparison of MR findings between MS patients, mutations carrier vs. non-carrier, matched for sex, age and disease duration, showed that the total number of lesions and the number of hypointense lesions on T1-weighted images was greater in MS patient carrying the ASA-PD mutations. Our results on genotype-phenotype correlation analysis indicate a possible contribution of detected arylsulfatase A gene polymorphisms to the clinical severity of multiple sclerosis, estimated by EDSS, MSSS and MR findings. The MSSS proved to be more appropriate indicator of disease progression and should be more frequently used in clinical practice especially for comparison of disease progression in different groups of patients and identification of factors that may influence disease progression such as the presence of gene polymorphisms. PMID- 21648306 TI - Gaze differences in processing pictures with emotional content. AB - The International Affective Picture System (IAPS) is a set of standardized emotionally evocative color photographs developed by NIMH Center for Emotion and Attention at the University of Florida. It contains more than 900 emotional pictures indexed by emotional valence, arousal and dominance. However, when IAPS pictures were used in studying emotions with the event-related potentials, the results have shown a great deal of variation and inconsistency. In this research arousal and dominance of pictures were controlled while emotional valence was manipulated as 3 categories, pleasant, neutral and unpleasant pictures. Two experiments were conducted with an eye-tracker in order to determine to what the participants turn their gaze. Participants were 25 psychology students with normal vision. Every participant saw all pictures in color and same pictures in black/white version. This makes 200 analyzed units for color pictures and 200 for black and white pictures. Every picture was divided into figure and ground. Considering that perception can be influenced by color, edges, luminosity and contrast and since all those factors are collapsed on the pictures in IAPS, we compared color pictures with same black and white pictures. In first eye-tracking IAPS research we analyzed 12 emotional pictures and showed that participants have higher number of fixations for ground on neutral and unpleasant pictures and for figure on pleasant pictures. Second experiment was conducted with 4 sets of emotional complementary pictures (pleasant/unpleasant) which differ only on the content in the figure area and it was shown that participants were more focused on the figure area than on the ground area. Future ERP (event related potential) research with IAPS pictures should take into consideration these findings and to either choose pictures with blank ground or adjust pictures in the way that ground is blank. For the following experiments suggestion is to put emotional content in the figure area and to use different non complementary pictures to see if there is difference between different emotional categories. PMID- 21648307 TI - Dendritic morphology and spine density is not altered in motor cortex and dentate granular cells in mice lacking the ganglioside biosynthetic gene B4galnt1 - A quantitative Golgi cox study. AB - Gangliosides are characteristic plasma membrane constituents of vertebrate brain used as milestones of neuronal development. As neuronal morphology is a good indicator of neuronal differentiation, we analyzed how lack of the ganglioside biosynthetic gene Galgt1 whose product is critical for production of four major adult mammalian brain complex gangliosides (GM1, GD1a, GD1b and GT1b) affects neuronal maturation in vivo. To define maturation of cortical neurons in mice lacking B4galnt1 we performed a morphological analysis of Golgi-Cox impregnated pyramidal neurons in primary motor cortex and granular cells of dentate gyrus in 3, 21 and 150 days old B4galnt1-null and wild type mice. Quantitative analysis of basal dendritic tree on layer III pyramidal neurons in the motor cortex showed very immature dendritic picture in both mice at postnatal day 3. At postnatal day 21 both mice reached adult values in dendritic length, complexity and spine density. No quantitative differences were found between B4galnt1-null and wild type mice in pyramidal cells of motor cortex or granular cells of dentate gyrus at any examined age. In addition, the general structural and neuronal organization of all brain structures, qualitatively observed on Nissl and Golgi Cox, were similar Our results demonstrate that neurons can develop normal dendritic complexity and length without presence of complex gangliosides in vivo. Therefore, behavioral differences observed in B4galnt1-null mice may be attributed to functional rather than morphological level of dendrites and spines of cortical pyramidal neurons. PMID- 21648308 TI - Cognitive abilities and language comprehension in preschool children with perinatal brain lesion. AB - Perinatal brain lesion is a risk factor for development, making parents of such children particularly worried about consequences it may have on the child's cognitive and language development. Although literature findings on the outcome of perinatal brain lesion are inconsistent, most of the studies have found a positive general outcome, but also subtle deficits that affect the child's academic success. Since language comprehension and cognitive abilities influence learning abilities at school, we wanted to know how six-year olds who were selected based on pathological ultrasonographical findings (ischemic or hemorrhagic brain lesion) would perform on subtests of Wechsler battery (WISC) and language comprehension measures (Reynell Developmental Language Scale and Peabody Picture Vocabulary Test), compared with controls. The second issue we investigated was whether in children who suffered a perinatal brain lesion cognitive abilities predicted the level of language comprehension in the same way as in children without perinatal brain lesion. The relation between cognitive and linguistic abilities is still a controversial one, and a different relation would mean that these two groups of children have different structure of abilities probably due to perinatal brain lesion. Forty children who suffered a perinatal brain lesion and forty age-matched children without perinatal risk factors were examined. Our results showed that the groups differed more in linguistic than in cognitive variables. Also, the two groups showed different relation patterns between cognitive abilities and language comprehension. Cognitive abilities were statistically significantly associated with language comprehension in children who suffered a perinatal brain lesion, while this association was not statistically significant within the control group. Since a number of participants with perinatal brain lesion had language difficulties, it is presumed that they rely on cognitive abilities in order to overcome and compensate for language shortcomings. PMID- 21648309 TI - Correlation of cognitive functions with some aspects of illness, treatment and social functioning in recurrently hospitalized schizophrenic patients. AB - Cognitive deficits are found to be contributors to poorer psychosocial functioning, rehabilitation outcome and lack of treatment success in schizophrenia. Aim of the study was to examine correlation of cognitive functions with some aspects of illness, treatment and social functioning in a group of recurrently hospitalized schizophrenic patients (N=60). Deficient results on psychomotor processing speed, verbal fluency and verbal learning correlated with the longer duration of illness, higher number of hospitalizations and shorter duration of regular antipsychotic treatment. Deficient results on verbal fluency correlated with the younger age of onset, poor functional autonomy and organizational skills, whereas deficient results on psychomotor processing and verbal learning correlated with poor organizational skills alone. Score on verbal fluency was predictive of social skills impairment, whereas score on psychomotor processing was predictive of functional autonomy and organizational skills impairment. Functioning of different cognitive domains could be predictive of functioning in different social domains. Interplay of specific cognitive deficit and social functioning could be responsible for recurrent hospitalizations and unfavorable treatment choices. PMID- 21648310 TI - The application of ultrasound in neuroendoscopic procedures: first results with the new tool "NECUP-2". AB - In this paper, our experience with originally constructed Neurosurgical Endoscopic Contact Ultrasound Probe "NECUP-2" in neuroendoscopy is reported. Between June 1997 and June 2007, 132 neuroendoscopic procedures have been performed: 102 endoscopic thrid ventriculostomies (ETV), 15 arachnoid cysts and 5 intraventricular tumours operations. The "NECUP-2" was applied effectively in all cases in which blunt perforation was not possible: 38/102 ETY, 10/10 septostomies, 15/15 arachnoid cysts. In five cases of intraventricular tumours, neuroendoscopic procedure was combined with open microsurgery for tumour removal with preservation of vascular structures. There were no "NECUP-2" related complications. Of postoperative complications, we had liquorrhea (9 patients), and symptoms of meningitis (6 patients). In the follow-up period (6 months to 6 years), we had a patency rate of 80% (50/63 patients). All patients improved in clinical status. According to the first results, it seems that ultrasonic contact probe NECUP-2 presents a new device in neurosurgical armamentarium that can be used in various fields of neurosurgery. With minimal and controlled lesion that is produced at the tip of the probe, it can be used in highly demanding operations such as third ventriculostomy and tumour resection. PMID- 21648311 TI - Physical characteristics in the new model of the cerebrospinal fluid system. AB - It is unknown which factors determine the changes in cerebrospinal fluid (CSF) pressure inside the craniospinal system during the changes of the body position. To test this, we have developed a new model of the CSF system, which by its biophysical characteristics and dimensions imitates the CSF system in cats. The results obtained on a model were compared to those in animals observed during changes of body position. A new model was constructed from two parts with different physical characteristics. The "cranial" part is developed from a plastic tube with unchangeable volume, while the "spinal" part is made of a rubber baloon, with modulus of elasticity similar to that of animal spinal dura. In upright position, in the "cranial" part of the model the negative pressure appears without any measurable changes in the fluid volume, while in "spinal" part the fluid pressure is positive. All of the observed changes are in accordance to the law of the fluid mechanics. Alterations of the CSF pressure in cats during the changes of the body position are not significantly different compared to those observed on our new model. This suggests that the CSF pressure changes are related to the fluid mechanics, and do not depend on CSF secretion and circulation. It seems that in all body positions the cranial volume of blood and CSF remains constant, which enables a good blood brain perfusion. PMID- 21648312 TI - Effects of vibrotactile stimulation on the control of muscle tone and movement facilitation in children with cerebral injury. AB - Afferent signals from the muscle's proprioceptors play important role in the control of muscle tone and in the facilitation of movements. Peripheral afferent pathway enables the restoration of connections with supraspinal structures and so includes mechanism of synaptic inhibition in the performance of normal movement. Different sensory stimuli, as vibrotactile stimulation, excite muscle's proprioceptors which then send sensorimotor information via spinal cord. In this way afferent signals promote cortical control and modulation of movements. The goal of this study is to evaluate the effects of vibrotactile stimulation on the spasticity and motor performance in children with cerebral injury. Subjects included in this study were 13 children who were developing the classification of spastic cerebral palsy. For all children perinatal brain damage was documented by medical reports and neonatal brain ultrasound scan. At the mean age of 3 years and 6 months subject underwent the assessment of motor development by Gross Motor Function Measurement (GMFM-88). Gross Motor Classification System (GMFCS) has been used to classify functions of lower extremities. Therapeutic intervention was conducted once a week during 3 months. All subjects were stimulated with vibrotactile stimuli of 40Hz in duration of 20 minutes in order to reduce spasticity. After the ending of the treatment subjects underwent second assessment of motor performance and the classification of lower extremities functions. The results have shown that there was a significant improvement in motor performance, what has been seen in the facilitation of rotations, better postural trunk stability and head control and in greater selectivity of movements. Further randomized, control trial investigations with bigger sample and included spasm scale are needed to gain better insight in the role of vibrotactile stimulation in the facilitation of normal movements. PMID- 21648313 TI - Writing and drawing with both hands as indicators of hemispheric dominance. AB - Brain lateralization is a common term used to describe dominance of one brain hemisphere over another for a specific function. The right hand dominance in writing, controlled by the left hemisphere, is preceded by development of communicative gesticulation and followed by development of speech in the same hemisphere. We assumed that some people are not aware of their own capability of using the other hand for tasks involving fine motor sequential movements. To prove this hypothesis, the participants were asked to perform one trained task (writing) and one less-trained task (drawing) with a dominant and a non-dominant hand. The final sample was comprised of 1189 children from 14 elementary schools and 8 high schools in the Osijek area, of which 685 elementary school children were attending 1st to 4th grade and 504 high school children were attending 3rd and 4th grade. The participants were asked to write two words, draw a specific object (a vase with flowers) and fill out a questionnaire with 10 questions concerning the classification of handedness and cerebral hemisphere dominance. The self-reported cerebral lateralization assessed in the questionnaire was compared with the drawing and the writing performance. The self-reported and objectively measured hand dominance deviated in the cases of the ambidextrous who consider themselves right-handers. Given the fact that the number of ambidextrous persons was greater in elementary schools than in high schools, we concluded how training of the right hand decreases the ability of using both hands equally for either of the tested functions - writing and drawing. PMID- 21648314 TI - Potential error in ventriculocisternal perfusion method for determination of cerebrospinal fluid formation rate in cats. AB - The cerebrospinal fluid (CSF) formation rate (Vf) has been extensively studied by the ventriculocisternal perfusion, a method still regarded as the most precise one. This method as well as the equation for the calculation of the CSF formation rate (Vf) was established by Heisey et al on indicator dilution in perfusate. They assumed that the dilution of the indicator substance in perfusion is a consequence of newly formed CSF i.e. a higher CSF formation rate would result in a higher degree of dilution of the indicator substance. Therefore, such method is indirect and any mistake in the interpretation of the degree of indicator dilution would lead to questionable and often contradictory results regarding CSF formation rates. According to Heisey's equation, Vf shoud not depend on the rate of ventriculocisternal perfusion. However it has been shown that Vf is perfusion dependt value, and also that during perfusion the indicator substance is partially absorbed into surrounding tissue. It is possible that obtained Vf dependence on perfusion rate was caused by observed absorption of indicator substances. For that reason, in anaesthetised cats ventriculocisternal perfusion was performed at higher (252.0 microL/min) and at lower perfusion rate (65.5 microL/min) and Vf was calculated at both experimental and corrected (just for absorbed amount) values of indicator substance. Since (inspite of the correction) the difference of 12.4 microL/min between lower (15.0 microL/min) and higher perfusion rate (27.4 microL/min) was obtained, it is obvious that ventriculocisternal perfusion method cannot be considered reliable for measuring CSF formation rate. PMID- 21648315 TI - Association study of a functional catechol-o-methyltransferase polymorphism and cognitive function in patients with dementia. AB - A functional catechol-o-methyltransferase (COMT Val158/108Met) polymorphism, a valine (Val) to methionine (Met) substitution, has been associated with cognitive processing in the normal brain, older age, mild cognitive impairment and in various dementias. COMT is involved in the breakdown of dopamine and other catecholamines, especially in the frontal cortex; hence the carriers of Met allele, with the lower enzymatic activity, are expected to perform better on particular neuro-cognitive tests. The study included 46 patients with dementia and 65 healthy older subjects. The neurological status was assessed, using the Mini Mental Status Examination (MMSE), and the batery of different neurological tests. In DNA samples COMT polymorphism was genotyped. Patients with dementia exhibited significant genotype-induced differences in scores for MMSE, Visual Association Test (VAT) duration of numbers test, VAT time of response to numbers test, VAT average response to numbers test and WPLCR/PPLR unanswered. Carriers of Met/Met genotype had significantly lower scores of MMSE, significantly longer time to respond to VAT duration of numbers test, VAT time of response to numbers test and VAT average response to numbers test, and significantly greater number of unanswered questions to WPLCR/PPLR when compared to Met/Val or Val/Val genotypes. Our preliminary data showed significantly impaired performance in several neuro-cognitive tests in carriers of Met/Met genotype in patients with dementia compared to either Met/Val or Val/Val genotype carriers. Although Met/Met genotype with more dopamine available in the frontal cortex should be associated with better neuro-cognitive test results than Met/Val or Val/Val genotype, our data on patients with dementia did not confirm this hypothesis. Further study on larger sample of patients is needed to clarify the role of COMT polymorphism in cognitive functions. PMID- 21648316 TI - Effect of olive oil on brain's lipid and calcium content after partial hepatectomy in mice. AB - The aim of this study was to investigate the influence of olive oil (00) enriched diet on the lipid content of mice brain during the early phase of liver regeneration and to test a relationship of these changes with calcium content. C57BI mice were fed over 21 days with diet enriched with olive oil, containing predominantly oleic acid (18:1n-9). The animals were one-third partially hepatectomised (pHx) under aether anaesthesia. Total lipids were extracted from tissue samples with a chloroform-methanol (2:1, v/v) mixture according to Folch et al. Mineral concentration was determined by inductively coupled plasma atomic emission spectroscopy (ICP-AES) after microwave brain tissue digestion. The diet containing 00 increased both total lipid content and the calcium concentration in brain during the early phase of liver regeneration (12hrs post pHx), suggesting that monounsaturated oleic acid might interact with some metal-dependent activities that control changes in the brain during liver regeneration. PMID- 21648317 TI - Rat sciatic nerve crush injury and recovery tracked by plantar test and immunohistochemistry analysis. AB - An experimental crush injury to the sciatic nerve, with a crush force of 49.2 N (pressure p=1.98x10(8) Pa), was inflicted in 30 male rats (Wistar). A control group (sham), with the same number of rats, was also operated upon exactly as the experimental group but without the crush injury. We tested the sensory and motor recovery of the sciatic nerve with Hargreaves method, using an apparatus from Ugo Basile, Italy. Testing was continued for both legs of each rat, injured and uninjured, starting preoperatively (0 day), and then 1, 7, 14, 21, and 28 days postoperatively. The same experiment was run simultaneously with the sham group. The Plantar test showed recovery of the sensory and motor function of the sciatic nerve, though not complete recovery, by 28 days. An immunohistochemical experiment was run in parallel with the plantar test on L3-L6 segments of the spinal cord from where the sciatic nerve extends. We used antibodies for Myelin associated glycoprotein (MAG), and gangliosides GD1a and GT1b on the aforesaid part of the spinal cord. The immunohistochemical methods showed changes in sensory and motor axons in the spinal cord segment L3-L6 which suggest correspondence with the results of the Plantar test, in terms of recovery of the sensory and motor function after injury of the sciatic nerve. The immunohistochemical results also show ipsilateral and contralateral changes following injury. Results of the plantar test are suggestive that the rat shows compensation for an injury in its contralateral leg. PMID- 21648318 TI - AXIN-1 protein expression and localization in glioblastoma. AB - The etiology and pathogenesis of tumors of the central nervous system are still inadequately explained. In the present study the expression patterns of a critical molecular component of wnt signaling pathway - axin I was investigated in 42 patients with glioblastoma, the most aggressive form of glial tumors. Immunostaining and image analysis revealed the quantity and localization of the protein. Downregulation of this tumor suppressor expression was observed in 31% of tumors when compared to the levels of axin in healthy brain tissues. Axin was observed in the cytoplasm in 69% of glioblastoma samples, in 21.4% in both the cytoplasm and nucleus and 9.5% had expression solely in the nucleus. Mean values of relative axin's expression obtained by image analysis showed that the highest relative quantity of axin was measured when the protein was in the nucleus and the lowest relative quantity of axin when the protein was localized in the cytoplasm. Investigation on axin's existence at the subcellular level in glioblastomas suggests that axin's expression and spatial regulation is a dynamic process. Despite increasing knowledge on glioma biology and genetics, the prognostic tools for glioblastoma still need improvement. Our findings on expression of axin 1 may contribute to better understanding of glioblastoma molecular profile. PMID- 21648319 TI - Methods for measuring acoustic power of an ultrasonic neurosurgical device. AB - Measurement of the acoustic power in high-energy ultrasonic devices is complex due to occurrence of the strong cavitation in front of the sonotrode tip. In our research we used three methods for characterization of our new ultrasonic probe for neuroendoscopic procedures. The first method is based on the electromechanical characterization of the device measuring the displacement of the sonotrode tip and input electrical impedance around excitation frequency with different amounts of the applied electrical power The second method is based on measuring the spatial pressure magnitude distribution of an ultrasound surgical device produced in an anechoic tank. The acoustic reciprocity principle is used to determinate the derived acoustic power of equivalent ultrasound sources at frequency components present in the spectrum of radiated ultrasonic waves. The third method is based on measuring the total absorbed acoustic power in the restricted volume of water using the calorimetric method. In the electromechanical characterization, calculated electroacoustic efficiency factor from equivalent electrical circuits is between 40-60%, the same as one obtained measuring the derived acoustic power in an anechoic tank when there is no cavitation. When cavitation activity is present in the front of the sonotrode tip the bubble cloud has a significant influence on the derived acoustic power and decreases electroacoustic efficiency. The measured output acoustic power using calorimetric method is greater then derived acoustic power, due to a large amount of heat energy released in the cavitation process. PMID- 21648320 TI - Serum lipid levels in patients with Alzheimer's disease. AB - The role of lipids in the aetiology and progress of Alzheimer's disease (AD) is still unclear High lipid levels could be one of the risk factors for AD, but no association or even protective effects of high cholesterol levels in the development of the AD were also found. The aim of the study was to determine serum levels of total cholesterol, high-density-lipoprotein cholesterol (HDL-C), low-density-lipoprotein cholesterol (LDL-C) and triglycerides (TG) in female patients with AD and in healthy elderly controls. The 50 patients met the diagnostic criteria of probable AD according to the NINDS-ADRDA and DSM-IV criteria. Cognitive impairment was evaluated using the Mini Mental State Examination (MMSE). Patients were subdivided into two groups of 19 patients in the middle (MMSE 10-19) and 31 patients in the late (MMSE 0-9) phase ofAD. Psychotic and non-psychotic features, evaluated by means of Neuropsychiatric Inventory, were presented in 13 and 37patients with AD, respectively. Control group consisted of 58 subjects without cognitive impairment (MMSE >27) and with lipid levels within normal range. Serum lipid levels were determined by the enzymatic colour tests and by the enzymatic clearance assay. Significantly lower lipid levels were found in patients with AD, than in controls. Patients in the late phase of AD had significantly lower entire lipid profile than controls and significantly lower cholesterol and LDL-C levels than patients in the middle stage ofAD. There was no difference in lipid levels between patients with and without psychotic features. The significant positive correlations were found between MMSE scores and cholesterol, LDL-C levels and age in all AD patients. The results support the presumption that lipid profile might be connected with the aetiology and progress of AD and showed the association between low serum cholesterol and LDL-C levels and cognitive decline in patients with AD. Further studies are needed to confirm the relationship between lipid levels and cognition, and to validate the lipid profile as a biological marker for the progress of AD. PMID- 21648321 TI - Distribution of major brain gangliosides in olfactory tract of frogs. AB - Gangliosides are major cell-surface determinants in the central nervous system (CNS) of vertebrates, found both in neuronal and glial cell membranes. Together with cholesterol and glycosylphosphatidylinositol (GPI) - anchored proteins, gangliosides are involved in organization of plasma membrane microdomains. Based on biochemical studies, frog brain was previously described as having low quantities of gangliosides and their distribution pattern in specific brain regions was unknown. Using highly specific monoclonal antibodies generated against four major brain gangliosides (GM1, GD1a, GD1b and GT1b), we examined the distribution of these molecules in CNS of four different species of frogs (Rana esculenta, Rana temporaria, Bufo bufo and Bufo viridis). We also studied the distribution of myelin- associated glycoprotein (MAG), an inhibitor of axonal regeneration, which is a ligand for gangliosides GD1a and GT1b. Our results show that ganglioside GDla is expressed in neurons of olfactory bulb in all studied animals. In the brain of Rana sp., GD1a is expressed in the entire olfactory pathway, from olfactory bulbs to amygdala, while in Bufo sp. GD1a is restricted to the main olfactory bulb. Furthermore, we found that most of myelinated pathways in frogs express MAG, but do not express GD1a, which could be one of the reasons for better axon regeneration of neural pathways after CNS injury in amphibians in comparison to mammals. PMID- 21648322 TI - Super-ego in patients with coronary artery disease. AB - In this paper, we explored the super-ego ofpatients with coronary artery disease. Research results have confirmed the initial hypothesis that a significant number of patients with coronary artery disease has rigorous super-ego. Among patients with coronary artery disease (N=50), and control group (N=50), we have found significant differences in the quality of super-ego and ego attitude towards the demands of the super-ego. The results of this research contribute to understanding the impact of psychological factors in coronary artery disease. PMID- 21648323 TI - Blood pressure dipping and salivary cortisol as markers of fatigue and sleep deprivation in staff anesthesiologists. AB - Anesthesiologists often work extended duty shifts that result in acute and chronic sleep loss and circadian disruption. Stress caused by sleep deprivation, together with excessive workload could contribute to acute increases in blood pressure (BP) and sympathetic nervous system activity. Non-dipping pattern of BP is considered an additional risk factor for cardiovascular events and target organ damage. We hypothesized that there would be significant changes of cardiovascular parameters when comparing work on call during the 24-hour in hospital shift (24-HD) versus ordinary working day (8-HD) combined with changes of dipping pattern and altered diurnal cortisol secretion, measured by salivary cortisol (SC). Following local Medical Ethics Committee approval, 12 out of 36 staff anesthesiologists (8 male, 4 female), 33-61 years old, participated in this study. Ambulatory BP monitor was used for noninvasive 24-hour ambulatory BP and heart rate (HR) monitoring. Each participant was monitored continuously during the 8-HD, as well as during the 24-HD. Saliva for analysis of cortisol levels was collected six times a day (at 8 am, 11 am, 2 pm, 5pm, 8pm, and 11 pm) both during 8-HD and on 24-HD. There was a significant decrease in number of diastolic dippers on call vs. diastolic dippers on ordinary working day (4/12 vs. 10/12, p=0.036), and non significant decrease of systolic dippers (3/12 vs. 7/12, p =0.214). There were no significant differences in SC values between 8-HD and 24 HD at all observed time points. However, the SC values measured during the night were markedly elevated on both days compared with reference values and the shapes of SC curves were altered. The lack of diastolic BP dipping could be more sensitive indicator of stress among staff anesthesiologists than systolic BP dipping. The shape of SC diurnal curve in terms of elevated night values could be another indicator of their chronic fatigue. PMID- 21648324 TI - Age-developmental stage and severity of trauma related symptoms, anxiety and depressive symptoms in participants who lost their fathers during the war in Croatia. AB - Children of different ages will experience a traumatic event in a different ways. The most important in the generalization of research findings is recognizing that children of different ages think differently, act differently and have different emotional functioning. Experiences that are extremely traumatic to an adult may be perceived by a young child as something that is not so frightening. The fear that the child feels will more frequently be a reflection of that of the adult rather than generated by the child's own perception of the event. So, the individual experience of the trauma is age dependent. Our study focused on children who lost their fathers in conditions of war The aim was to explore the association between age-developmental stages and the severity of trauma related symptoms, anxiety and depressive symptoms in participants who lost their fathers during the war. The study included 103 people who lost their fathers during the war in Croatia, who came to the physical and psychiatric examination organized by the Ministry of Family, War Veterans and Intergenerational Solidarity. The sample was consisted of the participants who were children, or not born yet, at the time when they lost their fathers during the war in Croatia. At the time of interview, the participants were aged between 15 and 35 years old. Data was collected using a structured clinical interview which also included socio-demographic data. Data about former and current psychiatric symptoms were collected using the following instruments: Clinician- Administrated PTSD Scale (CAPS), Hamilton anxiety scale (HAMA), Hamilton depression scale (HAMD). Results showed that there was significant correlation between age and results on used scales. The participants who lost their fathers at a very young age or even before they were born showed less trauma symptoms (r=0.249; p < 0.05) less anxiety (r=0.374; p < 0.01) and depressive (r=0.384; p<0.01) symptoms than participants who lost their fathers at an older age. The study confirmed that the individual experience of the trauma of losing a father in war circumstances is associated with age. PMID- 21648325 TI - Changes in brain metabolites measured with magnetic resonance spectroscopy in antidepressant responders with comorbid major depression and posttraumatic stress disorder. AB - In a present pilot study, performed on 11 subjects, we studied proton magnetic resonance spectroscopy (1H-MRS) changes in early to intermediate (3-6 weeks) responders to antidepressant treatment with selective serotonin reuptake inhibitors (SSRIs). All subjects had diagnosis of major recurrent depression comorbid to posttraumatic stress disorder (PTSD). Magnetic spectroscopy was done in the region of dorsolateral prefrontal cortex on a 3T MRI-unit. Participants were selected out of the larger sample due to an early response to antidepressant treatment within 3-6 weeks, measured with Beck Depression Inventory (BDI). We measured levels of neuronal marker N-acetyl-aspartate (NAA), choline (CHO) and creatine (Cr). There was no difference in NAA/Cr ratios between the first and the second spectroscopic scans (p= 0.751). However, CHO/Cr ratios showed increasing trend with mean value at the first scan of 1.09 (SD =0.22) while mean value at second scan was 1.25 (SD=0.24), displaying statistically significant difference (p=0.015). In conclusion, significant increase in choline to creatine ratio from the first to the second spectroscopic scan during the antidepressant treatment, compared to almost identical values of NAA to creatine ratio, suggests increased turnover of cell membranes as a mechanism of the early response to the antidepressant drug therapy. PMID- 21648326 TI - Lenticulostriatal vasculopathy - a marker for congenital cytomegalovirus infection? AB - Lenticulostriate vasculopathy (LSV) is an ultrasound (US) visible lesion of the brain, which appears as echogenic streaks or spots in the arteries of thalamus and basal ganglia. LSV has varied etiology. Transfontanelar Color Doppler (TFCD) can easily display lenticulostriatal blood flow and assess: stage I LSV with present flow within echogenic changes and stage II LSV in which the flow disappears, despite a presence of streaks and spots, which at this stage most probably correspond to calcification. The objectives of this study are to determine: (1) Whether there are differences in distribution (unilateral or bilateral) and presence (during first year of age) of TFCD flow between congenital CMV infection positive and negative group of children with LSV (2) Could US and TFCD findings of LSV be an indication for further investigation of possible congenital CMV infection, because of their variable and often adverse neurodevelopmental outcome? We examined and followed-up 98 infants with LSV One group (37/98) with congenital CMV infection and second (61/98) negative. All infants had clinical signs of neuromotor delay and ultrasound and TFCD markers of LSV Our study shows that most of the patients from both groups had TFCD visible flow at the age of 0-4 months. In majority of them in both groups, at the age of 5-8 months, there was no more visible flow. TFCD showed no statistically significant difference among congenital CMV infection positive group and negative group, nor in youngest age period (0-4 months), nor in later course of flow in LSV unilaterally or bilaterally. Although the LSV presents nonspecific marker for intracranial infection (ICI), all infants presenting with LSV should be evaluated for possible ICI. Thus, the Doppler findings of LSV in infants require a detailed examination, monitoring and follow-up of neuromotor outcome. PMID- 21648327 TI - Characteristics of the hearing loss in unilateral cleft lip and palate-influence on communication. AB - Etiology of otitis media with effusion (OME) is still unclear and often described as multi-factorial. It is very usual finding in cleft palate population. We tested relationship between the hearing level, audiometric frequencies, aging and ear side in unilateral cleft lip and palate 101 children (UCLP) and subgroups of left (UCLP)(L) and right cleft side (UCLP)(R). Group of left ears is prone to higher frequency and more severe hearing disturbances than groups of right ears, with less chance of normalizing hearing level with aging. Characteristics of hearing loss level and its improvement, in UCLP children depend of cleft type, ear side and age group. PMID- 21648328 TI - 1-H MRS changes in dorsolateral prefrontal cortex after donepezil treatment in patients with mild to moderate Alzheimer's disease. AB - Magnetic resonance spectroscopy (MRS) noninvasively provides information on the concentration of some cerebral metabolites in vivo. Among those measurable by proton magnetic resonance spectroscopy (1H-MRS), N-acetyl-aspartate (NAA) is decreased, and myo-inositol (ml) and choline (Cho) levels are increased in patients with Alzheimer's disease (AD). Donepezil, an acetylcholinesteraze inhibitor, has proven effect on cognitive symptoms in patients with AD. In previous studies, treatment response was associated with an increase of NAA and NAA/Cr in the parietal lobe and hippocampi. Correlation of longitudinal changes of 1H-MRS detectable metabolites in dorsolateral prefrontal cortex (DLPFC) with clinically observable changes is a poorly researched topic. The objective of this non-interventional study is to assess whether changes in 1H-MRS measurable metabolites correlate with clinical outcome after donepezil treatment. Twelve patients with mild to moderate AD were evaluated during 26 weeks of donepezil treatment. 1H-MRS parameters in DLPFC were assessed before and after 26 weeks of donepezil treatment. Cognition was assessed with Alzheimer's Disease Assessment Scale cognitive subscale (ADAS-Cog). A significant increase in NAA/Cr ratio and significantly lower decrease in mI/Cr ratio were found in AD patients with positive treatment response. The results of this study indicate possible modest donepezil effect on prevention of neuronal functional deterioration in DLPFC which correlates with clinical outcome and point the use of 1HMRS as technique of help in assessment of drug effect. PMID- 21648329 TI - Comparison of emotion recognition from facial expression and music. AB - The recognition of basic emotions in everyday communication involves interpretation of different visual and auditory clues. The ability to recognize emotions is not clearly determined as their presentation is usually very short (micro expressions), whereas the recognition itself does not have to be a conscious process. We assumed that the recognition from facial expressions is selected over the recognition of emotions communicated through music. In order to compare the success rate in recognizing emotions presented as facial expressions or in classical music works we conducted a survey which included 90 elementary school and 87 high school students from Osijek (Croatia). The participants had to match 8 photographs of different emotions expressed on the face and 8 pieces of classical music works with 8 offered emotions. The recognition of emotions expressed through classical music pieces was significantly less successful than the recognition of emotional facial expressions. The high school students were significantly better at recognizing facial emotions than the elementary school students, whereas girls were better than boys. The success rate in recognizing emotions from music pieces was associated with higher grades in mathematics. Basic emotions are far better recognized if presented on human faces than in music, possibly because the understanding of facial emotions is one of the oldest communication skills in human society. Female advantage in emotion recognition was selected due to the necessity of their communication with the newborns during early development. The proficiency in recognizing emotional content of music and mathematical skills probably share some general cognitive skills like attention, memory and motivation. Music pieces were differently processed in brain than facial expressions and consequently, probably differently evaluated as relevant emotional clues. PMID- 21648330 TI - Could a neurological disease be a part of Mozart's pathography? AB - As expected, since we recently celebrated the 250th anniversary of birth of Wolfgang Amadeus Mozart, there has been again a renewal of interest in his short but intensive life, as well as in the true reason of his untimely dead. Mozart lived and died in time when the medical knowledge was based mostly on subjective observations, without the established basics of standardized medical terminology and methodology. This leaves a great space for hypothesizing about his health problems, as well as about the cause of his death. The medical academic community attributed to Mozart approximately 150 different medical diagnoses. There is much speculation on the possible causes of Mozart's death: uremia, infection, rheumatic fever, trichinellosis, etc. Recently some authors have raised the question about a possible concomitant neurological disease. According to available records, Mozart has shown some elements of cyclotimic disorder, epilepsy and Gilles de la Tourette syndrome. Furthermore, the finding of a temporal fracture on (allegedly) Mozart's skull, gives a way to speculations about the possibility of a chronic subdural hematoma and its compressive effect on the temporal lobe. Despite numerous theories on Mozart's pathography that also include a concomitant neurological disorder, the medical and history records about Mozart's health status indicate that he probably had suffered from an infective illness, followed most likely by the reactivation of rheumatic fever, which was followed by strong immunologic reaction in the last days of his life. Taking all the above into consideration, it is reasonably to conclude that Mozart's neurological disturbances were caused by the intensity of the infective disease, and not primarily by a neurological disease. PMID- 21648331 TI - Correlation of sleep disturbances, anxiety and depression in Croatian war veterans with posttraumatic stress disorder. AB - The aim of the study was to examine the relationships between global sleep quality and its specific components and Posttraumatic Stress Disorder (PTSD) symptom severity questionnaire. We also researched whether sleep quality and sleep disturbances differed among groups of PTSD based on symptom severity categories. This study was conducted on the sample of 120 Croatian war veterans with PTSD. The following self-report instruments were used: Pittsburgh Sleep Quality Index, the Pittsburgh Sleep Quality Index Addendum for PTSD, the Mississippi Scale for Combat-Related PTSD, the Spielberger State and Trait Anxiety Inventory and the Beck Depression Inventory. There were statistically significant differences between the three PTSD severity groups on general nervousness (PSQI-A variable), where patients with extremely severe PTSD have more symptoms of general nervousness than groups with severe or moderate PTSD. Differences were found between PTSD severity groups in episodes of terror and acting-out dreams, where patients with extremely severe PTSD have more symptoms of episodes of terror and acting-out dreams than groups with severe or moderate PTSD. Sleep quality was significantly correlated with state anxiety, trait anxiety, and depression, indicating that with decrease of anxiety and depression, sleep quality improves. Sleep latency was positively correlated with both state and trait anxiety. There wasn't any significant correlation between sleep latency and depression. Study suggests that sleep disturbances are equally severe across groups of veterans based on PTSD severity and that the severity of sleep disturbances is significantly related to severity of anxiety and depression symptoms. PMID- 21648332 TI - Succinylcholine use in adult anesthesia - A multinational questionnaire survey. AB - There are no definitive evidence based standards regarding use of succinylcholine (SCh) for anesthesia induction. However, there is a global trend toward eliminating SCh not only in elective, but also in emergency surgery in adults. The aim of the study was to survey the use of SCh in adult elective and emergency anesthesia practice in several European countries and the United States by questionnaire. One hundred and seventy anesthesiologists out of 201 possible, from six institutions in five countries (Croatia, Bosnia and Herzegovina, Hungary, United Kingdom, and the United States) anonymously completed the questionnaire about their use of SCh. The questionnaire was structured to assess the respondents': frequency of use of SCh in adult surgery (elective and emergency), reasons for use or rejection of SCh, positive and negative attributes of SCh, and observed side effects in their practice. Differences in use were tested using X2-test when appropriate. There was a significant difference in the use of SCh between countries for elective surgery with the lower use in UK and Hungary (chi2=45.8, p <0.001). One hundred and seventeen (69%) use it regularly. In emergency surgery 165 (97%) anesthesiologists use it without any significant difference among countries (chi2=2.13, p<0.711). The top indications for SCh use were anticipated difficult intubation/ventilation (74%), caesarean section (54%), and obesity and/or hiatus hernia (49%). The top reasons against SCh use were adequate substitutes (87%), fear of arrhythmias (45%), and anaphylaxis (19%). The most desirable reported drug features were: rapid onset (88%), short duration (64%), and effective relaxation (61%). Forty-six per cent of the surveyed anesthesiologists stated they had never experienced a complication with its use. The most frequently reported side effects were myalgias (47%), bradycardias (42%), and prolonged blockade (39%). Allergic reactions were reported by 13%, and asystole by 12% of physicians. From our survey it is possible to conclude that succinylcholine is still regularly used, at least by surveyed anesthesiologists in Europe and USA, in adult anesthesia practice, especially in elective surgery for which it may be least suited. This reflects the discrepancies between the international guidelines for the use of SCh and the clinical practice of many anesthesiologists in different countries. The regional differences in SCh usage may be considered through anesthesia cultures and practice variations depending on country. PMID- 21648333 TI - Spatial and temporal measurements of eye movement in children with dyslexia. AB - This paper presents the first reading data in Croatian collected with an eye tracking device. The eye-tracking method allows for research into two crucial levels underlying reading: the visual and the cognitive. The aim of this paper is to show the differences in eye movements in children with dyslexia using the principles of cognitive-control view. Despite the well-known definitions and vast literature on dyslexia, the neural basis of dyslexia varies greatly on the individual level. The three children studied in this paper were tested behaviorally using set of language tests for language behavior assessment on all language levels: phonology, morphology, syntax, lexicon and pragmatics. Two children had low scores on most language tests, and all three children had poor reading and writing level. Each of the children had to read two texts silently while their eye movements were recorded by means of an infrared eye-tracking system. We analyzed the number, position, and duration of fixations and the number and position of regressive (or back) saccades. Our results show intergroup differences (between a typically developing child and the three children with dyslexia), and intragroup differences (among all three children with dyslexia). The great number of fixations, longer duration of fixations, and great number of regression saccades are the main features that differentiate the children with dyslexia form the typically developing child. The only difference found between language and visual subtypes of dyslexia was a shorter duration of fixations for the child with a visual processing disorder. PMID- 21648334 TI - Self-assessment of well-being as an indicator of quality of life of former war prisoners - A Croatian study. AB - The impact of war on the population is vast, especially when it comes to those who were directly affected by war, among other things as concentration camp detainees. Because of the specific war experience of this population it is important to better understand the possible contribution of key socio-demographic variables, war traumatization and acute disturbances in mental health to their subjective assessment of their own well-being, which represents a psychological category and is based on a subjective assessment. The starting point is a theoretical precept according to which individual characteristics, together with war experience, can have repercussions on mental health, and eventually on the general well-being of an individual and their quality of life. The study comprised 184participants who had given their informed consent for participation and filled out complete questionnaires. The participants were a convenience sample of male persons who had survived war captivity in the Homeland War in the period from 1991 to 1995. The study was conducted as part of the physical examinations at the University Hospital "Fran Mihaljevic" in Zagreb. The data was collected using several self-evaluation measuring instruments one of which served to collect socio-demographic data, two to collect data on the participants' mental health, one for the data on the participants' combat and war experiences and one to assess the participants' well-being. The data obtained suggest that only avoidance and arousal symptoms and psychosomatic difficulties are predictors of the well-being of persons who have experienced war captivity. PMID- 21648335 TI - Subjective assessment of quality of life of persons whose fathers had died in the homeland war. AB - The aim of this paper is to show the current subjective assessment of the quality of life of persons whose fathers had died in war, and compare it with regard to some of their socio-demographic characteristics. The study included 494 participants who had come for a physical and psychiatric examination to one of the health institutions in Zagreb, Rijeka, Osijek or Split in which examinations were arranged. The inclusion criterion was growing up in a single-parent family as a consequence of the father dying in war. Data were collected using a structured clinical interview which also included socio-demographic data: the age and gender of the participant, educational status, marital status, employment status, household income. Also, participants were asked to fill out the World Health Organization Quality of Life Questionnaire - short form (WHOQOL-BREF). Data obtained from this study are descriptive, and are in line with the data obtained on the general population when it comes to comparisons of the assessment of quality of life and certain demographic characteristics. Special emphasis should be given to the link between age and overall satisfaction with the quality of life, with younger persons being more satisfied with the total quality of life than the older study participants. In conclusion, all participants of this study had a specific traumatic experience during the war - their father's death, therefore more data regarding the quality of life of this population can be expected upon a more detailed analysis and establishment of the contribution of traumatization, socio-demographic variables and current mental health to the explanation of the subjective satisfaction with life. A more detailed analysis of the collected data will be available in subsequent papers. PMID- 21648336 TI - Infantile spasms in children with Down syndrome. AB - Down syndrome (DS) is the most common genetic cause of mental retardation. It is estimated that 5-13% of persons affected by DS have seizures. Infantile spasms are the most common type of seizures and usually are well controlled with steroids and antiepileptic drugs. We present 11 children at the age of 3 years and 4 months to 10 years and 7 months with DS and infantile spasms, treated at Children's Hospital Zagreb from January 2000 until July 2009. Infantile spasms began at the age of 5 to 10.5 months in 10 children, in one child at the age of 16 months. Only one child had perinatal risk factors for the development of IS. Changes in EEG correlated to hypsarrhythmia. Infantile spasms were treated initially with antiepileptic drugs, most often with valproic acid. Treatment was inefficient in 10/11 patients. After application of ACTH, infantile spasms stopped between 7 and 15 days in 6 patients, until 28th day in 4 patients. Hypsarrhythmia vanished in all children. During follow-up period (2 years and 7 months to 9 years and 5 months) none of the children developed another type of seizures. No major epileptogenic changes were registered in EEG. Antiepileptic therapy was discontinued in 4 children (aged 4 years and 2 months to 5 years). In this group is the boy who died of heart failure. Infantile spasms associated with DS are categorized into symptomatic group. The existence of cerebral pathology and delayed psycho-motor development precedes occurrence of seizures. It is possible to achieve good control of seizures and disappearance of hypsarrhythmia with application of ACTH and antiepileptic drugs. PMID- 21648337 TI - Serum cholesterol concentration and structured individual psychoanalytic psychotherapy in suicidal and non-suicidal male patients suffering from borderline personality disorder. AB - Findings from numerous studies suggest an association between low cholesterol levels and suicidal behavior in patients with different psychiatric diagnoses. The aims of this case-control study were to test whether cholesterol levels in male suicidal patients (N=20) with borderline personality disorder (BPD) are lower than in male non-suicidal patients (N=20) with BPD and male healthy control group (N=20), and to evaluate the influence of structured individual psychoanalytic psychotherapy on suicidal behavior. The groups were matched for age and body mass index (BMI). Results showed that serum cholesterol levels did not differ significantly between suicidal and non-suicidal BPD patients and healthy controls. The level of psychopathology (measured by Brief Psychiatric Rating Scale and Hamilton Depression Rating Scale) was significantly higher in the group of suicidal patients, which indicates the importance of evaluating particular clinical symptoms in BPD, in order to prevent suicidal behavior. Non suicidal male patients suffering from BPD received more frequently structured individual psychoanalytic psychotherapy prior to the hospitalization than suicidal group. These results emphasized the role of this type of psychotherapy in preventing suicidal behavior in BPD patients. PMID- 21648338 TI - Anxiety and depression in pregnant women with previous history of spontaneous abortion. AB - Spontaneous abortion is the most common adverse pregnancy outcome, and stress has been suggested to be important factor. We hypothesized that those female pregnant women with previous spontaneous abortion will have higher anxiety and depression than female pregnant women without previous spontaneous abortion and controls (healthy non-pregnant women). Age and socio-demographic parameters did not differ significantly between the three groups of participants. Anxiety and depression levels were assessed with the Beck Depression Inventory and the Beck Anxiety Inventory. There were no significant differences in the scores on Beck Depression Inventory between three groups of participants at the week 10 of gestation-1st assessment and week 20 of gestation-2nd assessment. Contrary of these results, pregnant women with history of spontaneous abortion have had statistically significant higher anxiety score than pregnant women without history of spontaneous abortion and control group-non-pregnant women, on both assessment of anxiety. Negative correlations between months passed form the last spontaneous abortion and anxiety on both assessment, emphasize the role of psychological support for the women who have experienced spontaneous abortion. PMID- 21648339 TI - Malformations of cortical development in children with congenital cytomegalovirus infection - A study of nine children with proven congenital cytomegalovirus infection. AB - Congenital cytomegalovirus (CMV) infection is the most common vertically transmitted disease with the rate of the infection ranging from 0.2 to 2.4% in newborn infants. Congenital CMV infection causes multiorgan affection, but the most severe and permanent sequelae are those affecting central nervous system such as mental retardation, cerebral palsy, sensorineural hearing loss, chorioretinitis and seizures as a result of direct interference of the virus with neurogenesis. The time of acquiring infection is strongly connected to the level of child's disability. Infection in early pregnancy results in severe neurological sequelae, while later infection has less prominent signs. Radiological findings show connection between onset of infection and brain imaging, from lissencephaly, pachygyria, polymicrogyria, schizencephaly, calcification, cerebellar hypoplasia and/or hypoplasia/agenesis of corpus callosum as a result of an early infection, to white matter abnormalities including disturbed myelination as a result of a late infection. We present nine patients with proven congenital CMV infection and malformations of cortical development and their computed tomography/magnetic resonance (CT/MRI) findings along with clinical assessments. According to CT/MRI results we assume that two of our children with lissencephaly had an early onset of infection. The other seven with less severe cortical dysplasia in form of pachy/polymicrogyria were probably infected later Cerebellar hypoplasia and/or calcifications in our patients also confirm an early onset of infection. Developmental outcome in all of our children was poor: moderate to severe psychomotor retardation has been diagnosed in all children; five of them have developed cerebral palsy (four have bilateral spastic and one dyskinetic) and one is estimated to have minor motor dysfunction. Seven out of nine developed epilepsy, chorioretinitis was found in three of them and sensorineural deafness in two of them. All of our children, except one, were presented by symptomatic infection, yet only four of them were recognized at birth. Therefore, congenital CMV infection should be considered as one of the reasons for childhood disability more often. PMID- 21648340 TI - A validation study of appropriate phonological verbal fluency stimulus letters for use with croatian speaking individuals. AB - The aim of this study is to determine the word frequency for all thirty letters of the Croatian alphabet and to collect normative data for the letter fluency task in Croatian speakers. Ninety two healthy participants were given each of the Croatian letters, and asked to generate as many words as possible in 60 seconds for each letter Results suggested that participants generated most frequently words starting with the letters as follows: "K", "P", "S" and "M". PMID- 21648341 TI - Cognitive auditory evoked potentials in children with special language impairment. AB - Perception and discrimination of auditory and speech stimuli in children aged 7-9 years with either receptive (n=6) or expressive (n=5) type of special language impairment and 7 healthy age-matched controls was investigated using evoked potential technique. The measurements were performed with a 32-channel Neuroscan electroencephalographic system. Two types of stimuli were applied, pure tones (1 kHz and 2 kHz) and double syllabi consisting of one consonant and one vocal characteristic of Croatian language. The stimuli were presented in an oddball paradigm, requiring a conscious reaction for the subjects. Latencies and amplitudes of P1, N1, P2, N2, P3, N4, and SW waves were analized, as well as the reaction time and number of responses. There were found no statistically significant difference between children with special language impairment and the control group in average response time and number of responses to tone burst or double syllable. Analysis of variance of all used variables showed a statistically significant difference in P3 and Sw wave latencies after double syllable stimulation, P3 and N4 waves latencies after target stimulation, P2 and Sw wave amplitude; and in N1 wave amplitude after pure tone stimulation. Our study showed that children with speech and language disorder take longer time to perceive and discriminate between either tonal or speech auditory stimuli than children with typical speech and language development. PMID- 21648342 TI - Comparison of hippocampal volumes in schizophrenia, schizoaffective and bipolar disorder. AB - The reduction of hippocampal volume was frequently reported in schizophrenia, but not in bipolar disorder This volume reduction is associated with clinical features of schizophrenia, in particular with working and verbal memory impairments. Schizoaffective disorder, as a specific disorder sharing clinical features of both schizophrenia and bipolar disorder is rarely analyzed as a separate disorder in neurobiological studies. The aim of this study was to compare hippocampal volumes in separate groups of patients with schizophrenia, schizoaffective and bipolar disorder. Hippocampal volumes were estimated using high resolution magnetic resonance imaging in 60 subjects, 15 subjects in each patient and one healthy volunteer (control) group. There were no significant differences in hippocampal volume between bipolar disorder and control group. Hippocampal volume was statistically significantly reduced in the group of patients with schizophrenia and schizoaffective disorder, compared to either bipolar disorder or control group, thus supporting the hypothesis that hippocampal volume reduction could be considered as a possible neurobiological basis for clinical aspects of schizophrenia and schizoaffective disorder associated with working and verbal memory impairment. PMID- 21648343 TI - Effect of environmental enrichment on morphology of deep layer III and layer V pyramidal cells of occipital cortex in oldest-old rat - A quantitative golgi cox study. AB - Dendrites and dendritic spine density regress extensively during aging in rats housed under standard conditions (SC), which can be ameliorated by housing in the enriched environment (EE). This event is particularly pronounced on neurons where high rates of plasticity are conceivable, such as on projection neurons of archicortical regions of dentate gyrus'. However, effects of EE on neocortical projection neurons are still poorly understood. Therefore, we investigated the effect of EE housing on a deep layer III (L3) and layer V pyramidal cell (L5) morphology in the associative occipital neocortex of male Sprague-Dawley rats at 24 months of age. Rats were randomly distributed in two groups and reared under either SC (n=5) or EE conditions (n=6) for 26 days. In depth quantitative analysis of dendritic tree morphology and spine density on occipital projection neurons, from Golgi-Cox stained sections, showed similar trend in both EE occipital layers L3 and L5. Significant increase was found in total number of dendritic segments (L3 - 37.5 %, L5 - 33 %) and in dendritic diameter of intermediate segments (for more than 20 %), while increase in total spine number was around the level of significance (p>0.55; L3 - 30 %, L5 - 64 %). These findings suggest an outgrowth of new dendritic segments, When compared to archicortical region of dentate gyrus, effects of aging in the associative occipital cortex were less pronounced. Taken together, these findings suggest that structures being more affected by the aging process are more susceptible to the environmental enrichment in old age. PMID- 21648344 TI - Hepatic and pancreatic glycosphingolipid phenotypes of the neurological different rat strains. AB - Among three commonly used strains of laboratory rats, Wistar rats perform more neurological tasks better then Lewis and Sprague-Dawley (SD) rats. Liver is the main site of insulin-like growth factor (IGF) production and pancreas is the exclusive site of insulin production. Insulin stimulates neuronal development and appropriate IGF-I input is critical in brain growth. Glycosphingolipids (GSLs) are important mediators of insulin secretion and action. Therefore, this study investigated GSL phenotypes of liver and pancreas with hypothesis that they are different in three rat strains. Total GSL fractions (neutral and gangliosides) were analysed by high performance thin-layer chromatography (HPTLC). Complex gangliosides were detected by HPTLC immunostaining using cholera toxin B subunit after neuraminidase pretreatment. Wistar rats had the highest liver weight/body weight ratio and SD rats had the highest pancreas weight/body weight ratio. Ganglioside GM3 was more expressed in the liver of Wistar compared to Lewis and SD rats. SD rats contained scarce quantities of GD1a and b-series gangliosides in the liver compared to Wistar and Lewis rats. Pancreatic b-series ganglioside content was also the lowest in SD rats. This study represents differences in the hepatic and pancreatic ganglioside phenotypes of three rat strains that could influence IGF and insulin secretion and action. PMID- 21648345 TI - Aggresiveness in institutionalised schizophrenic patients and the selection of antipsychotics. AB - The selection of antipsychotics as medications used primarily for treating schizophrenia and disorders similar to schizophrenia is an important aspect of the treatment of forensicpatients. This study examines the effect of antipsychotics selection (typical or atipycal) on the level of aggressiveness, side effects and the hospitalisation length. The research is conducted on 98 psychiatric patients diagnosed with schizophrenia or similar disorders (F 20-F 29) in two forensic psychiatric institutions. The patients committed aggressive criminal offence in state of insanity. The patients are currently treated in inpatient psychiatric institutions. The research was conducted by using the Aggressiveness Questionnaire (AG-87), the Simpson-Angus Scale for the assessment of extrapyramidal side effects, the Barnes Akathisia Rating Scale for the assesment of akathisia and the Abnormal Involuntary Movement Scale. The results show no significant difference between the groups of patients treated with typical and atypical antipsychotics in all the variables. PMID- 21648346 TI - EEG polysomnographic study of maturational differences between twins. AB - The purpose of this pilot study is to asses the effects of multiple pregnancies on the maturation of the developing brain using the polysomnographic EEG recordings. Data from prospectively recorded 10 twin sets, born prematurely (mean 36 gestational week; range 33 - 38 GW) in the Split University Hospital Center, were analysed. We compared sleep architecture parameters in the twins at 37h and 44th postmenstrual age (PMA) with parameters that were expected at that PMA. The same parameters were compared within each twin pair using the Man Whitne test. At first measurement indeterminate sleep (IS) proportion was greater in the first twin than in the second one. The IS sleep proportion was 1.6 fold greater in the first twin (p=0.028), and 1,8 fold less percentage of quite sleep (QS) than the second twin (p=0.054). The length of sleep stages among the twins was similar at the second measurement. Measures of sleep architecture were not significantly different within the twins in second recording. The results of this study obtained on a relatively small number of twins (longer IS and shorter QS in the first twin at the 38th week recordings), showed that the maturational differences among twins exist in utero and shortly after birth, and then disappear until the end of the first month of the postnatal life. PMID- 21648347 TI - Hydroxyapatite ceramics in multilevel cervical interbody fusion - is there a role? AB - The aim of this study is to evaluate the efficacy of hydroxyapatite grafts in multilevel cervical interbody fusion during the one year follow-up. A total of 86 patients with degenerative cervical disc disease underwent all together 224 cervical interbody fusion procedures in which either Smith-Robinson or Cloward type hydroxyapatite grafts were used. The surgeries included radiculopathy in 38 cases, myelopathy in 20 cases and myeloradicuopathy in 28 patients. In 65 out of 86 patients, fusion was followed by an anterior instrumentation (plating). Postoperatively, patients were followed for a mean of 15.64 (range 11-23.3) months. All patients underwent radiography to evaluate fusion and the axis curvature. Excellent clinical results (86%), described as a complete or partial relief of symptoms with full return to preop activity, were obtained in patients with radiculopathy. There were 5 grafts mobilizations and one graft fracture. Two grafts extruded in non-instrumented patients and required repeated surgery. There were other three reoperations due to the hardware problems. One year fusion rate was obtained at 86% for two-level surgery, 80.1% for three-level surgery and 74% for four-level surgery. The mean (SD) hospital stay was 3.8 (0.7) days. A hydroxyapatite cheramic can be a very effective synthetic material for multilevel cervical interbody fusion. It is characterized by a high fusion rate and a small percentage of graft-related complications, especially when fusion procedure is followed by plating. PMID- 21648348 TI - Quality of life in families of Croatian veterans 15 years after the war. AB - Exposure to war trauma with its consequences such as post traumatic stress disorder (PTSD) and disability due to combat injuries poses a significant problem for modern Croatian society. However, this is also a public health problem requiring continuous study of effective treatment strategies to achieve an increase in quality of life of most war-affected groups. Aim of this study was to examine the quality of life of population most affected by war - families of Croatian veterans. Present study included 126 female participants, who agreed to complete physical and psychiatric examination organized by the Ministry of Family, War Veterans and Intergenerational Solidarity. Included were participants with status of either wife of war veteran suffering from PTSD, wife who lost her husband in war circumstances or wife of war veteran with physical disabilities resulting from war activities. All three groups were asked to fill out the World Health Organization Quality of Life Questionnaire - short form (WHOQOL-BREF). Results indicate that assumed intensity of secondary trauma is not associated with quality of life. Namely, the highest level of satisfaction was found in wives of the most seriously affected invalids of war (M=3.77; sd=0.741), folowed by the wives of deceased soldiers (M=3.5; sd= 0.697), while the lowest quality of life results were found in wives of veterans suffering from PTSD (M=3.12; sd=0.608). Our results confirm that, nearly 15 years after the war, wives of disabled or killed Croatian soldiers have a (comparatively) satisfactory quality of their everyday lives, compared to the wives of veterans suffering from PTSD. PMID- 21648349 TI - Differential diagnostic relevance of high resolution magnetic resonance in patients with possible multiple system atrophy (MSA) - A case report. AB - Multiple system atrophy (MSA) is sporadic, progressive neurodegenerative disorder characterized clinically by autonomic dysfunction, Parkinsonism (MSA-P), and cerebellar ataxia (MSA-C) in any combination. Parkinsonism is present in the majority of patients (80%). Early in the course of the disease autonomic dysfunctions are present in approximately 40% of patients, while the domination of cerebellar symptoms is present in 20% of all patients. According to second consensus statement on diagnosis of MSA, to make the diagnosis of possible MSA, except Parkinsonism or a cerebellar syndrome, there must be one feature involving autonomic dysfunction plus one other additional that can include findings on history, clinical examination or changes in structural or functional imaging. We present a case of 60-year old male with Parkinsonism and cerebellar symptoms accompanied with signs of autonomic nervous system involvement. Level of autonomic dysfunction was not the level required for the diagnosis of probable MSA. On initially performed 1.5T MRI, the most prominent neurodegenerative feature of brain stem, cerebellum and basal ganglia was atrophy, however features like "hot-cross bun" sign, "slit-like" putaminal rim and middle cerebellar peduncle hyperintensities were detected only after MR imaging on higher resolution (3T) device. Our case points to the possibility that some typical structural changes that can help in diagnostic process may not be clearly visible on 1.5 T MRI devices. In such cases we suggest using 3T MRI device, if feasible, in order to demonstrate findings that may help in establishing the diagnosis of possible MSA. PMID- 21648350 TI - Misdiagnosis and exacerbation of unusual obsessive-compulsive disorder presentation with risperidone and clozapine in an adolescent girl - A case report. AB - Obsessive-compulsive disorder (OCD) is a heterogenous disorder with different clinical presentations. The most common symptoms are those that involve contamination, possible harm, ordering/symmetry, aggressive/sexual/religious concerns and hoarding. A variety of less common symptoms have been described. Unusual OCD symptoms may lead to misdiagnosis, inappropriate treatment with possible serious side effects. In this report we present a case of an adolescent girl in which unusual OCD presentation and symptoms were misinterpreted to represent psychosis and exacerbation of OCD symptoms with risperidone and clozapine treatment. We discuss the possible pathophysiological mechanisms of OCD symptom exacerbation, clinical implications, and successful management of this case, with fluvoxamine therapy. This case may represent the first report of musical obsessions successfully managed with fluvoxamine therapy. PMID- 21648351 TI - The treatment of autistic children with risperidone. AB - Autism is a pervasive developmental disorder characterised by impairment in social interaction and communication, with unusual behavior. In some cases the pharmacotherapy is prescribed and the most studied antipshychotic drugs include haloperidol and risperidone. In this paper we displayed the treatment of two cases of autism in boy and girl with risperidone. With the use of risperidone in girl, we have achieved reduction of psychomotor symptoms and reduction of hetero aggressive and self-destructive behavior, and in boy we have also achieved reduction of psychomotoric symptoms, with improvement in contact with his surrounding, he had less learning problems and he has felt familiar not only with his mother, but with other persons. Research on the use of risperidone in the treatment of autistic disorders among children in Croatia are rare, given the limited use of risperidone in children younger than 15years, the question arises about the need to expand the scope of application of risperidone in younger age groups. PMID- 21648352 TI - Coexistance of cerebral sinovenous thrombosis and Dandy Walker malformation in newborn. AB - Cerebral sinovenous thrombosis in neonatal period may cause neurological impairment, epilepsy, and lead to stroke. It is caused primarily by coagulopathy of numerous reasons, occasionally perinatal asphyxia, traumatic delivery and hyperhomocysteinemia. Dandy-Walker malformation is characterized by agenesis or hypoplasia of the cerebellar vermis, cystic dilatation of the fourth ventricle, and enlargement of the posterior fossa. Dandy-Walker malformation, variant, and mega cisterna magna represent a spectrum of developmental anomalies. Insults to developing cerebellar hemispheres and the fourth ventricle are believed to be the cause of malformation. Our patient was born from noncomplicated pregnancy, noncomplicated nontraumatic vaginal delivery at term, excellent Apgar scores, without peculiarities in clinical status. She was brest-fed by the 42nd hour of life when she had rightsided seizures during sleep that repeated for five times in next 24 hours. Brain Ultrasound (US) revealed clot in left lateral ventricle, slight dilatation of left ventricle, both sided periventricular echodensity, ischemia, slight enlargement of forth ventricle and a bit smaller cerebellum. There was no visible flow through left transverse, superior sagittal and straight sinus. Magnetic Resonance (MRI) confirmed the finding and showed thrombosis of left and right transverse venous sinuses and confluence of sinuses. Electroencephalogram (EEG) showed leftsided focal changes. The newborn was treated with phenobarbiton for 8 days and had no convulsions during that period. All coagulation parameters, homocistein, lipoproteins (a) and D-dimers were normal. There were no mutations on FV R506Q, PT 20210A, MTHFR 677C/T. No antiphospholipides were found. Heart US showed no structural anomalies. No other patology or risk factors were present at the time. Before discharge, US showed hydrocephalus. Flow in affected sinuses was visible with color Doppler. MRI showed recanalization of affected sinuses, also hydrocephalus and presentation of Dandy Walker On EEG there was borderline finding. Due to progression of hydrocephalus ventriculo-peritoneal shunt was placed. In age of 1 year EEG was slower for age but without focus. Neurological development was normal for age. The question is whether this child had intrauterine insult and inception of Dandy Walker with further postnatal progress of thrombosis and evolution to full picture of Dandy Walker with hydrocephalus OR thrombosis that led to development of hydrocephalus and Dandy Walker malformation in this child were accidental coexistance. PMID- 21648353 TI - Optimizing the function of upstanding activities in adult patients with acquired lesions of the central nervous system by using the Bobath concept approach - A case report. AB - Nonspecific medical gymnastic therapy may help patients after stroke achieve certain results in terms of efficiency but not in terms of quality of movement. The goal of treatment by Bobath concept is development of movement (effectiveness) and optimization of movement (efficiency). This article presents the case of a 62-year old patient who had experienced a stroke and has difficulties with standing up activities. It underscores the importance of not only recovery of function but also optimization of the function in such patients. PMID- 21648354 TI - Tremor, seizures and psychosis as presenting symptoms in a patient with chronic lyme neuroborreliosis (LNB). AB - Lyme borreliosis is a multisystem disorder caused by Borrelia burgdorferi (Bb). Neurological symptoms such as lymphocytic meningoradiculoneuritis (Bannwart's syndrome), cranial neuritis (II,III,IV,V,VI), encephalitis, transverse myelitis are found in about 10% of cases during the second phase of the disease. In the chronic stage, many months or years after the initial infection, other neurologic complications may occur, such as encephalomyelitis, epileptic crises, cognitive impairment, peripheral neuropathy and psychiatric disturbances such as depression, anxiety, panicc attacks, catatonia, psychosis etc. Some patient continue to experience symptoms of fatigue, insomnia or psychiatric disorder in the post borrelia syndrome. We describe here a patient with a triad of unusual symptoms in chronic LNB including tremor, seizures and psychosis. Standardized medical interview, neurologic examination, neuroimaging, serum and CSF serology as well as EEG and EMNG evaluation were performed. The patient was treated with intravenous ceftriaxone and doxycycline and responded with rapid clinical and functional improvement.Newertheless, he suffered from multiple systemic and neurologic sequelas that influenced his daily activities in post treatment period. Emphasis is placed on the atypical onset and evolution, the difficulties encountered in formulating diagnosis, early treatment and the uncertainties concerning the sequelae after treatment. In patients with non-specific long lasting symptoms in the absence of overt clinical signs suggesting CNS involvement, routine treatment with i.v. ceftriaxone is not to be encouraged. PMID- 21648355 TI - Coexistence of intradural spinal arteriovenous malformation and associated developmental anomalies - report of two cases. AB - Spinal arteriovenous malformations (AVM) have been devided into dural (Type I), intramedullary glomus (Type II), juvenile (Type III), and perimedullary direct arteriovenous fistulae (Type IV). AVMs are usually associated with subacute myelopathy in what has been known as Foix-Alajouanine syndrome. We presented two patients with two intradural spinal arteriovenous malformations associated in what we call Foix-Alajouanine syndrome. The both patient developed acute back pain and paresthesias, followed by paraplegia and incontinence. The clinical status of one patient has been improved after particle embolization for a 17 years when he deteriorated up to paraplegia after spinal angiography for follow up. Clinical status in another patient deteriorated, because particle emoblisation cannot be performed due to very descrete presentation of the feeding artery. Extensive neuroradiological examination in both patients revealed coexistence of numerous associated developmental anomalies in both patients. We conclude that arteriovenous malformations occasionally are associated with other vascular and nonvascular developmental anomalies elsewhere in the body. These findings rise attention about keep in mind the suspicion of mutual etiopathogenesis and congenital origin of these anomalies. Early timing of the diagnostic and therapeutic interventiosn are stressed to prevent or delay irreversible ishaemic myellopathy or haemorrhage. For the definitive diagnosis of spinal arteriovenous malformations and evaluation of its occlusion grade after the therapy spinal angiography is needed PMID- 21648356 TI - Fragile X-premutation tremor/ataxia syndrome (FXTAS) in a young woman: clinical, genetics, MRI and 1H-MR spectroscopy correlates. AB - It is generally thought that fragile X-associated tremor/ataxia syndrome (FXTAS) represents a late-onset neurodegenerative disorder occuring in male carriers of a premutation expansion (55-200 CGG repeats) in the fragile X mental retardation 1 (FMR 1) gene. However, several female patients with FXTAS have also been reported recently. Here, we describe a 23-year old woman with positive family history of mental retardation and autism who presented clinically with action tremor, ataxia, emotional disturbances and cognitive dysfunction. Magnetic resonance imaging (MRI) of the brain showed diffuse cortical atrophy, while 1H-MR spectroscopy (MRS) revealed decreased levels of N-acetylaspartate (NAA) in the cerebellum, basal ganglia, and pons. Genetic testing confirmed heterozygous FMR 1 gene premutation of 100 CGG repeats in the abnormal allele and 29 CGG repeats in the normal allele. We concluded that FXTAS may be an under-recognized disorder, particularly in women. PMID- 21648357 TI - Symptomatic capillary telangiectasia of the pons and intracerebral developmental venous anomaly - A rare association. AB - Various combinations of vascular malformations of the brain in one lesion have been reported, while others seem to be very rare. In this report, the authors discuss the case of a coexistence of an capillary telangiectasia of the pons and intracerebral venous anomaly. To our knowledge, this is the first report of coexistence of a capillary telangiectasia of the pons and intracerebral venous anomaly apparted from each other. These discrete vascular malformations of the brain raise attention on possible interrelations in the pathogenesis of these entities. We report a case of pontine capillary telangiectasia and intracerebral venous anomaly in a 42-year-old woman with a right side facial palsy. Hight field magnetic resonance imaging suggested presence of a capillary telangiectasia of the pons. Another lesion in the left frontal gyrus was attributable to the venous anomaly. Along with neuroradiological findings, results of the somatosensor evoked potentials, brain stem auditory potentials, laboratory analysis including blood, cerebrospinal fluid and urine investigation are demonstrated. Awareness of the magnetic resonance imaging finding of the capillary telangiectasias and of the venous anomalies may help in defining clinical correlates of this vascular malformations, while the follow up of these malformations might help to asses risk of vascular rupture. We and others previously selects capillary telangiectasia and venous anomaly in two discrete entities. Coexistence of these malformations in the brain apparted from each other appear to be very rare and raise attention on possible interactions in their natural history and pathogenesis. PMID- 21648358 TI - Aggressive intestinal schwannoma malignum mimicking gynecological pathology - A case report. AB - Primary malignant schwannoma of the small and large intestine is an extremely rare disease. Therefore, we are going to report an aggressive multifocal malignant intestinal schwannoma in a 66-year old female patient, that was primarily diagnosed as the gynecological tumor that, even after the surgical treatment, had a very quickly recurrence. Small intestine tumors may show images similar to an adnexal tumor, so it is difficult to differentiate one from another prior to the surgery. The patient did not suffer from neurofibromatosis type 1 (NF-1), disease that increases occurrence of malignant schwannoma in comparison with general population. These tumors are often diagnosed late, and radical surgical intervention does not guarantee longer survival. After surgical removal of macroscopically visible tumor masses from this patient, tumor formation within one month after the operation had reached the sizes of 83x66 mm and 85x75 mm respectively, with the occurrence of metastases in the liver, and thereafter the patient died. In differential diagnosis of adnexal tumor small intestine tumor has to be considered, especially if nonspecific symptoms are present. PMID- 21648359 TI - Jelena Krmpotic-Nemanic (1921-2008): contributions to human neuroanatomy. AB - Jelena Krmpotic-Nemanic (1921-2008) was a world-famous anatomist, internationally distinguished otolaryngologist, a member of the Croatian Academy of Sciences & Arts and appreciated professor at the School of Medicine University of Zagreb. The founding influence in her scientific career came from her mentor Drago Perovid who was a student of Ferdinand Hochstetter, the leading authority in the field of human developmental neuroanatomy and embryology. Such an influence was obviously important in early shaping of the research agenda of Jelena Krmpotic Nemanic, and it remains important in a long series of studies on developing human telencephalon initiated by Ivica Kostovic and his collaborators - with an always present and active support of Jelena Krmpotic-Nemanic. The aim of this mini review is to briefly describe her numerous contributions to the anatomy of the human peripheral and central nervous system. PMID- 21648385 TI - Microwave spectrum and conformational composition of 1-vinylimidazole. AB - The microwave spectrum of 1-vinylimidazole has been investigated in the 21-80 GHz spectral region. The spectra of two conformers have been assigned. One of these forms is planar, while the other is nonplanar with the imidazole ring and the vinyl group forming an angle of 15(4) degrees from coplanarity. The planar form is found to be 5.7(7) kJ/mol more stable than the nonplanar rotamer by relative intensity measurements. The spectra of 10 vibrationally excited states of the planar form and one excited-state spectrum of the nonplanar form were assigned. The vibrational frequencies of several of these states were determined by relative intensity measurements. The microwave work has been augmented by quantum chemical calculations at the CCSD/cc-pVTZ, MP2/cc-pVTZ, and B3LYP/cc-pVTZ levels of theory. The B3LYP calculations predict erroneously that both forms of 1 vinylimidazole are planar, whereas the MP2 and CCSD calculations correctly predict the existence of a planar and a nonplanar conformer of this compound. PMID- 21648386 TI - Specific photodynamics in thymine clusters: the role of hydrogen bonding. AB - A photoionization detected IR study of thymine and 1-methylthymine monohydrates and of their homodimers was carried out to shed some light on the structure of the thymine clusters whose complex photodynamics has recently been the subject of great interest. Under supersonic jet conditions, thymine forms doubly H-bonded cyclic clusters with water or another base preferentially via its N1-H group and the adjacent carbonyl group. This hydrate is of no biological relevance since the N1-H group is the sugar binding site in thymidine. On the other hand, 1 methylthymine forms the donor H-bonds only via the N3-H group. Hence, properties of the N1-H and the N3-H bound clusters of thymine can be studied using thymine and 1-methylthymine molecules, respectively. No biologically relevant conformations of the dimers and hydrates of thymine, contrary to those of 1 methylthymine, are observed under supersonic jet conditions. Thymine homodimer, which extensively fragments upon UV ionization by formation of a protonated monomer, exhibits two N1-H...O?C2 hydrogen bonds. The photodynamics of hydrated thymines is found to be extremely sensitive to the hydration site: ranging from an ultrafast relaxation in less than 100 fs up to formation of a dark state with the lifetime on the microsecond time scale. PMID- 21648387 TI - Dimer radical cations of indole and indole-3-carbinol: localized and delocalized radical cations of diindolylmethane. AB - Extending our previous study on the title species (J. Phys. Chem. A2010, 114, 6787), we investigated the dimer cations that are formed on oxidation of the glucobrassin derivatives indole-3-carbinol (I3C) and diindolylmethane (DIM) and of parent indole (I). Radiolysis in ionic liquid and Ar matrices shows that, at sufficiently high concentrations and/or on annealing the solid glasses, intense intermolecular charge-resonance (CR) absorption bands in the NIR herald the formation of sandwich-type dimer cations. The molecular and electronic structure of these species is modeled by calculations with the double-hybrid B2-PLYP-D density functional method which yields predictions in good accord with experiment. The radical cation of DIM also shows a CR band, but unlike in the case of I and I3C, its occurrence is not dependent on the concentration but instead on the solvent: in ionic liquid the CR band is initially absent and arises only on annealing, whereas in Ar matrices it is present from the outset and undergoes blue shifting and sharpening on annealing. These puzzling findings are rationalized on the basis of B2-PLYP-D calculations which predict that neutral DIM exists in the form of two conformers, present in different relative amounts in the two experiments, which on vertical ionization form distinct radical cations, a nonsymmetric one where the odd electron is largely localized on one of the two indole moieties and one with C(2) symmetry where charge and spin are completely delocalized over both halves of the molecule, thus giving rise to an intramolecular CR transition. On annealing, the nonsymmetric cation relaxes to a similarly delocalized structure with C(s) symmetry, thus explaining the observed increase and the shift of the CR band. We believe that DIM(*+) represents the first example of a radical cation which can exist under the same conditions as a localized and a delocalized complex cation. PMID- 21648388 TI - Time-resolved resonance Raman and computational investigation of the influence of 4-acetamido and 4-N-methylacetamido substituents on the chemistry of phenylnitrene. AB - A time-resolved resonance Raman (TR(3)) and computational investigation of the photochemistry of 4-acetamidophenyl azide and 4-N-methylacetamidophenyl azide in acetonitrile is presented. Photolysis of 4-acetamidophenyl azide appears to initially produce singlet 4-acetamidophenylnitrene which undergoes fast intersystem crossing (ISC) to form triplet 4-acetamidophenylnitrene. The latter species formally produces 4,4'-bisacetamidoazobenzene. RI-CC2/TZVP and TD B3LYP/TZVP calculations predict the formation of the singlet nitrene from the photogenerated S(1) surface of the azide excited state. The triplet 4 acetamidophenylnitrene and 4,4'-bisacetamidoazobenzene species are both clearly observed on the nanosecond to microsecond time-scale in TR(3) experiments. In contrast, only one species can be observed in analogous TR(3) experiments after photolysis of 4-N-methylacetamidophenyl azide in acetonitrile, and this species is tentatively assigned to the compound resulting from dimerization of a 1,2 didehydroazepine. The different photochemical reaction outcomes for the photolysis of 4-acetamidophenyl azide and 4-N-methylacetamidophenyl azide molecules indicate that the 4-acetamido group has a substantial influence on the ISC rate of the corresponding substituted singlet phenylnitrene, but the 4-N methylacetamido group does not. CASSCF analyses predict that both singlet nitrenes have open-shell electronic configurations and concluded that the dissimilarity in the photochemistry is probably due to differential geometrical distortions between the states. We briefly discuss the probable implications of this intriguing substitution effect on the photochemistry of phenyl azides and the chemistry of the related nitrenes. PMID- 21648389 TI - Effect of oxygen on the electronic structure of highly crystalline picene films. AB - The electronic structure of highly crystalline picene films with a standing-up orientation grown epitaxially on the Ag(110) surface was investigated. Upon exposure to oxgen gas, O(2) molecules incorporate at the interstitial sites within the a-b plane of the film. Features related to the highest three occupied molecular orbitals shift toward a lower binding energy which results in the inactivation of traps and the reduction of the charge injection barrier by about 1 eV. It is suggested that the highest two picene orbitals are inverted due to the strong interactions between the singly occupied oxygen pi orbital and the highest occupied orbital of picene. PMID- 21648390 TI - Templated construction of a Zn-selective protein dimerization motif. AB - Here, we report that the approach of metal-templated ligand synthesis can be applied to construct a dimeric protein assembly ((BMOE)RIDC1(2)), which is stabilized by noncovalent interactions and flexible covalent cross-linkers around the Zn templates. Despite its flexibility, (BMOE)RIDC1(2) selectively binds Zn(II) over other divalent metals and undergoes dimerization upon metal binding. Such simultaneous fulfillment of plasticity and selectivity is a hallmark of cellular signaling events that involve ligand/metal-induced protein dimerization. PMID- 21648391 TI - Suppression of inhomogeneous segregation in graphene growth on epitaxial metal films. AB - Large-scale uniform graphene growth was achieved by suppressing inhomogeneous carbon segregation using a single domain Ru film epitaxially grown on a sapphire substrate. An investigation of how the metal thickness affected growth and a comparative study on metals with different crystal structures have revealed that locally enhanced carbon segregation at stacking domain boundaries of metal is the origin of inhomogeneous graphene growth. Single domain Ru film has no stacking domain boundary, and the graphene growth on it is mainly caused not by segregation but by a surface catalytic reaction. Suppression of local segregation is essential for uniform graphene growth on epitaxial metal films. PMID- 21648392 TI - Redox potential dependence of peptide structure studied using surface enhanced Raman spectroscopy. AB - We describe a novel surface enhanced Raman spectroscopy (SERS) sensing approach utilizing modified gold nanoshells and demonstrate its application to analysis of critical redox-potential dependent changes in antigen structure that are implicated in the initiation of a human autoimmune disease. In Goodpasture's disease, an autoimmune reaction is thought to arise from incomplete proteolysis of the autoantigen, alpha3(IV)NC1(67-85) by proteases including Cathepsin D. We have used SERS to study conformational changes in the antigen that correlate with its oxidation state and to show that the antigen must be in the reduced state in order to undergo proteolysis. Our results demonstrate that a redox potential of ~ 200 mV was sufficient for reduction and subsequent productive processing of the antigenic fragment alpha3(IV)NC1(67-85). Moreover, we demonstrate that the peptide bonds subsequently cleaved by Cathepsisn D can be identified by comparison with a SERS library of short synthetic peptides. PMID- 21648393 TI - Radiation engineering of optical antennas for maximum field enhancement. AB - Optical antennas have generated much interest in recent years due to their ability to focus optical energy beyond the diffraction limit, benefiting a broad range of applications such as sensitive photodetection, magnetic storage, and surface-enhanced Raman spectroscopy. To achieve the maximum field enhancement for an optical antenna, parameters such as the antenna dimensions, loading conditions, and coupling efficiency have been previously studied. Here, we present a framework, based on coupled-mode theory, to achieve maximum field enhancement in optical antennas through optimization of optical antennas' radiation characteristics. We demonstrate that the optimum condition is achieved when the radiation quality factor (Q(rad)) of optical antennas is matched to their absorption quality factor (Q(abs)). We achieve this condition experimentally by fabricating the optical antennas on a dielectric (SiO(2)) coated ground plane (metal substrate) and controlling the antenna radiation through optimizing the dielectric thickness. The dielectric thickness at which the matching condition occurs is approximately half of the quarter-wavelength thickness, typically used to achieve constructive interference, and leads to ~20% higher field enhancement relative to a quarter-wavelength thick dielectric layer. PMID- 21648394 TI - Tuning structure and properties of graded triblock terpolymer-based mesoporous and hybrid films. AB - Despite considerable efforts toward fabricating ordered, water-permeable, mesoporous films from block copolymers, fine control over pore dimensions, structural characteristics, and mechanical behavior of graded structures remains a major challenge. To this end, we describe the fabrication and performance characteristics of graded mesoporous and hybrid films derived from the newly synthesized triblock terpolymer, poly(isoprene-b-styrene-b-4-vinylpyridine). A unique morphology, unachievable in diblock copolymer systems, with enhanced mechanical integrity is evidenced. The film structure comprises a thin selective layer containing vertically aligned and nearly monodisperse mesopores at a density of more than 10(14) per m(2) above a graded macroporous layer. Hybridization via homopolymer blending enables tuning of pore size within the range of 16 to 30 nm. Solvent flow and solute separation experiments demonstrate that the terpolymer films have permeabilities comparable to commercial membranes, are stimuli-responsive, and contain pores with a nearly monodisperse diameter. These results suggest that moving to multiblock polymers and their hybrids may open new paths to produce high-performance graded membranes for filtration, separations, nanofluidics, catalysis, and drug delivery. PMID- 21648395 TI - Peptide/laccase cocatalyzed asymmetric alpha-oxyamination of aldehydes. AB - An asymmetric alpha-oxyamination could be successfully performed by a peptide catalyst and laccase. The combination of peptide catalysis and enzymatic air oxidation promoted the reaction smoothly in water without employing a metal reagent. The oxyaminated compounds could be obtained as both aldehyde and carboxylic acid products depending on the reaction conditions. PMID- 21648396 TI - A concise route to dihydrobenzo[b]furans: formal total synthesis of (+) lithospermic acid. AB - A sequence of Sonogashira coupling, Pd(II)-catalyzed carbonylative annulation, and benzofuran reduction (Mg, MeOH, NH(4)Cl) provides a convergent and modular synthetic route to trans-2-aryl-2,3-dihydrobenzo[b]furan-3-carboxylates, which are a structural feature of numerous biologically active natural products. This versatile strategy was applied to the formal total synthesis of the anti-HIV natural product (+)-lithospermic acid. PMID- 21648397 TI - Excitation and emission spectra of jet-cooled naphthylmethyl radicals. AB - Gas phase excitation and emission spectra of three naphthylmethyl radical chromophores are presented. These resonance-stabilized species, 1-naphthylmethyl, 2-naphthylmethyl, and alpha-acenaphthenyl, each possessing an sp(2) carbon adjacent to a naphthalene moiety, are studied by resonant two-color two-photon ionization, laser induced fluorescence, and dispersed fluorescence spectroscopy. Identification of the radicals is made through a combination of dispersed fluorescence and density functional theory calculations. All three species possess spectra in the 580 nm region. The possible relevance to unidentified spectroscopic features such as the diffuse interstellar bands and emission from the Red Rectangle nebula is discussed. PMID- 21648398 TI - Syntheses of 5-formyl- and 5-carboxyl-dC containing DNA oligos as potential oxidation products of 5-hydroxymethylcytosine in DNA. AB - To investigate the potential oxidation products of 5-hydroxymethylcytosine (5 hmC)-containing DNA, we present here efficient syntheses of 5-formyl- and 5 methoxycarbonyl-2'-deoxycytidine phosphoramidites. The 5-formyl group in III was easy to introduce and was compatible with phosphoramidite and DNA syntheses. An additional treatment of ODN1 with NaBH(4) produced the corresponding ODN2 quantitatively. Phosphoramidite V was also incorporated into DNA, and the methyl ester could be hydrolyzed under mild basic conditions to afford ODN3. PMID- 21648399 TI - DFT and CASPT2 study on the mechanism of ethylene dimerization over Cr(II)OH+ cation. AB - In this work, the ethylene coordination and dimerization mechanism over Cr(II)OH(+) cation were systematically investigated using density functional theory (DFT) and complete active space second-order perturbation theory (CASPT2). It was found that Cr(II)OH(+) cation can coordinate with up to four ethylene molecules which gives seven possible stable Cr(II)OH(+).(C(2)H(4))(n) (n = 1-4) pi-complexes. We investigated whether ethylene dimerization over Cr(II)OH(+) cation proceeds through either a carbene mechanism or a metallacycle mechanism. The potential energy surfaces were characterized using four different functionals (M06L, BLYP, B3LYP, and M06). It was found that the potential energy profiles calculated at the M06 level agreed well with the CASPT2 energy profiles. Since the intermediates involved in the proposed catalytic cycles showed different ground spin states, a reaction pathway involving a spin crossing between two potential energy surfaces was observed. The minimum-energy crossing points (MECPs) that connect the two potential energy surfaces were successfully located. The two-state metallacycle reaction pathway with the formation of chromacyclopentane as the rate-determining step was found to be energetically more favorable than the carbene reaction pathway. 1-Butene was formed from the chromacyclopentane by a two-step reductive elimination pathway through a chromium(IV) hydride intermediate. PMID- 21648400 TI - Calorimetric study on coordination of tridentate imidazolyl calix[6]arene ligands to zinc ion in organic solvents. AB - Complexation of three kinds of tris(imidazolyl)calix[6]arenes containing alternate p-substituents (Calix-tBu, R(1) = R(2) = tert-butyl; Calix-NH(2), R(1) = tert-butyl, R(2) = NH(2); Calix-NO(2), R(1) = tert-butyl, R(2) = NO(2)) with Zn(ClO(4))(2)(H(2)O)(6) in acetonitrile, methanol, and THF was investigated via isothermal titration calorimetry (ITC). For the coordination of these calixarene ligands to Zn(II) in acetonitrile, typical one-phase exothermic titration curves were obtained, indicating the formation of 1:1 ligand-Zn(II) complexes accompanied by large conformational changes of the ligands. In contrast, the complexation in methanol was endothermic and dominated by favorable entropy changes. The entropy gains were achieved by extensive desolvation from both Zn(II) and the ligands. ITC measurements suggest a 2:1 ligand-Zn(II) complex formation in THF in the presence of excess ligands (Calix-NH(2) and Calix-NO(2)). The 2:1 complexes were converted to 1:1 complexes upon further addition of Zn(ClO(4))(2)(H(2)O)(6). The results indicate the important role of a coordinating solvent (acetonitrile) for direct formation of the 1:1 complexes under the conditions of excess ligand. Complexation of a ditopic ligand (Calix Tri) with three triazole moieties on the wider rim was also studied via ITC. The first coordination of the imidazole moieties to Zn(II) was an exothermic process. This was followed by the entropically favorable coordination of the triazole moieties to the divalent cation. We have also investigated exchange of the fourth ligand (H(2)O) of the Zn(II) complex of Calix-NH(2) with butylamine, heptylamine, acetonitrile, and acetamide in a noncompetitive solvent, THF. The DeltaH(0) tended to decrease upon increasing the electron-pair-donating ability of the guest ligand, whereas it was also affected by an entropic term due to restricted rotation of the guest ligand inside the calixarene cavity. PMID- 21648401 TI - Two new neptunyl(V) selenites: a novel cation-cation interaction framework in (NpO2)3(OH)(SeO3)(H2O)2 3H2O and a uranophane-type sheet in Na(NpO2)(SeO3)(H2O). AB - Dark green crystals of (NpO(2))(3)(OH)(SeO(3))(H(2)O)(2).H(2)O (1) have been prepared by a hydrothermal reaction of neptunyl(V) and Na(2)SeO(4) in an aqueous solution at 150 degrees C, while green plates of Na(NpO(2))(SeO(3))(H(2)O) (2) have been synthesized by evaporation of a solution of neptunyl(V), H(2)SeO(4), and NaOH at room temperature. Both compounds have been characterized by single crystal X-ray diffraction. The structure of compound contains three crystallographically unique Np atoms that are bonded to two O atoms to form a nearly linear O?Np?O NpO(2)(+) cation. Neighboring Np(5+) ions connect to each other through a bridging oxo ion from the neptunyl unit, a configuration known as cation-cation interactions (CCIs), to build a complex three-dimensional network. More specifically, each Np(1)O(2)(+), Np(2)O(2)(+), and Np(3)O(2)(+) cation is involved in three, five, and four CCIs with other units, respectively. The framework of neptunyl(V) pentagonal bipyramids is decorated by selenite trigonal pyramids with one-dimensional open channels where uncoordinated waters are trapped via hydrogen bonding interactions. Compound adopts uranophane-type [(NpO(2))(SeO(3))](-) layers, which are separated by Na(+) cations and water molecules. Within each layer, neptunyl(V) pentagonal bipyramids share equatorial edges with each other to form a single chain that is further connected by both monodentate and bidentate selenite trigonal pyramids. Crystallographic data: compound, monoclinic, P2(1)/c, Z = 4, a = 6.6363(8) A, b = 15.440(2) A, c = 11.583(1) A, beta = 103.549(1) degrees , V = 1153.8(2) A(3), R(F) = 0.0387 for I > 2sigma(I); compound (2), monoclinic, C2/m, Z = 4, a = 14.874(4) A, b = 7.271(2) A, c = 6.758(2) A, beta = 112.005(4) degrees , V = 677.7(3) A(3), R(F) = 0.0477 for I > 2sigma(I). PMID- 21648402 TI - Atom-economical chemoselective synthesis of 1,4-diynes and polysubstituted furans/pyrroles from propargyl alcohols and terminal alkynes. AB - Under different conditions, the reaction of propargyl alcohols and terminal alkynes leads to the selective formation of 1,4-diynes and polysubstituted furans/pyrroles. Water is the only byproduct in the selective synthesis of 1,4 diynes and pyrroles, and the strategy for the furan synthesis is of 100% atom economy. PMID- 21648403 TI - Palladium catalyzed C-C coupling reactions of 3,5-dichloro-4H-1,2,6-thiadiazin-4 one. AB - Palladium catalyzed Suzuki-Miyaura, Stille, and Sonogashira coupling reactions are reported for the electron-deficient heterocyclic scaffold 3,5-dichloro-4H 1,2,6-thiadiazin-4-one (1). Furthermore, 3,5-di(thien-2-yl)-4H-1,2,6-thiadiazin-4 one (7m) is further elaborated to afford the tetrathienyl 3,5-bis[(2,2'-bithien) 5-yl]-4H-1,2,6-thiadiazin-4-one (9). All compounds are fully characterized. PMID- 21648404 TI - Spirodiepoxide-based cascades: direct access to diverse motifs. AB - Allene epoxide formation/opening reaction sequences enabled direct access to diverse products. Described here are a single flask procedure for allene preparation and allene oxidation/derivatization reactions that give, among others, diendiol, diyndiol, alpha'-hydroxy-gamma-enone, dihydrofuranone, butenolide, and delta-lactone products. PMID- 21648405 TI - Moderately reactive molecules forming stable ionic compounds with superhalogens. AB - An explanation of the ability of selected moderately reactive molecules to form stable systems with superhalogens (AlF(4) and AlCl(4)) is provided on the basis of theoretical considerations supported by ab initio calculations. It is demonstrated that even the molecules possessing high ionization potentials (such as SiO(2), NH(3), CHCl(3), CCl(2)F(2)) should form stable and strongly bound ionic compounds when combined with a properly chosen superhalogen system (acting as an oxidizing agent). The conclusion is supported by providing the structural parameters and interaction energies for the SiO(2)AlF(4), NH(3)AlF(4), CHCl(3)AlF(4), CCl(2)F(2)AlF(4), SiO(2)AlCl(4), and NH(3)AlCl(4) compounds obtained at the CCSD(T)/6-311++G(d,p)//MP2/6-311++G(d,p) level. On the other hand, the AlF(4) and AlCl(4) superhalogens were found to be incapable of reacting with molecules whose ionization potentials (IP) exceed 13 eV (e.g., CO(2), CH(4)). Finally, it is demonstrated that the competition between the electron binding energy of the superhalogen system and the IP of the molecule the superhalogen is combined with is a key factor for predicting the stability of certain species. PMID- 21648406 TI - Uncoupling and antioxidant effects of ursolic acid in isolated rat heart mitochondria. AB - Ursolic acid (1), a pentacyclic triterpene acid, is one of the major components of certain traditional medicinal plants and possesses a wide range of biological effects, such as anti-inflammatory, antioxidative, and cytotoxic activities. Furthermore, 1, when present at 1.6-5 ng/mL concentrations in commercial herbal preparations used for patients with cardiac disorders, may also exert pro-cardiac activities. There are several indirect suggestions that the cardioprotective mechanism of ursolic acid could involve the mitochondria; however the mechanism of action is still not known. Therefore, the effects of 0.4-200 ng/mL ursolic acid (1) on the functions of isolated rat heart mitochondria oxidizing either pyruvate and malate, succinate, or palmitoyl-l-carnitine plus malate were investigated. It was found that 1 induced a statistically significant uncoupling of oxidative phosphorylation. A statistically significant decrease in H2O2 production in the mitochondria was observed after incubation with 5 ng/mL 1. This effect was comparable to the effectiveness of the classical uncoupler carbonyl cyanide 3-chlorophenylhydrazone. Since mild mitochondrial uncoupling has been proposed as one of the mechanisms of cardioprotection, the present results indicate that ursolic acid (1) has potential use as a cardioprotective compound. PMID- 21648407 TI - Bioorthogonal chemistry for site-specific labeling and surface immobilization of proteins. AB - Understanding protein structure and function is essential for uncovering the secrets of biology, but it remains extremely challenging because of the high complexity of protein networks and their wiring. The daunting task of elucidating these interconnections requires the concerted application of methods emerging from different disciplines. Chemical biology integrates chemistry, biology, and pharmacology and has provided novel techniques and approaches to the investigation of biological processes. Among these, site-specific protein labeling with functional groups such as fluorophors, spin probes, and affinity tags has greatly facilitated both in vitro and in vivo studies of protein structure and function. Bioorthogonal chemical reactions, which enable chemo- and regioselective attachment of small-molecule probes to proteins, are particularly attractive and relevant for site-specific protein labeling. The introduction of powerful labeling techniques also has inspired the development of novel strategies for surface immobilization of proteins to create protein biochips for in vitro characterization of biochemical activities or interactions between proteins. Because this process requires the efficient immobilization of proteins on surfaces while maintaining structure and activity, tailored methods for protein immobilization based on bioorthogonal chemical reactions are in high demand. In this Account, we summarize recent developments and applications of site-specific protein labeling and surface immobilization of proteins, with a special focus on our contributions to these fields. We begin with the Staudinger ligation, which involves the formation of a stable amide bond after the reaction of a preinstalled azide with a triaryl phosphine reagent. We then examine the Diels-Alder reaction, which requires the protein of interest to be functionalized with a diene, enabling conjugation to a variety of dienophiles under physiological conditions. In the oxime ligation, an oxyamine is condensed with either an aldehyde or a ketone to form an oxime; we successfully pursued the inverse of the standard technique by attaching the oxyamine, rather than the aldehyde, to the protein. The click sulfonamide reaction, which involves the Cu(I)-catalyzed reaction of sulfonylazides with terminal alkynes, is then discussed. Finally, we consider in detail the photochemical thiol-ene reaction, in which a thiol adds to an ene group after free radical initiation. Each of these methods has been successfully developed as a bioorthogonal transformation for oriented protein immobilization on chips and for site-specific protein labeling under physiological conditions. Despite the tremendous progress in developing such transformations over the past decade, however, the demand for new bioorthogonal methods with improved kinetics and selectivities remains high. PMID- 21648408 TI - Divergent synthesis of unsymmetrical annulated biheterocyclic compound libraries: benzimidazole linked indolo-benzodiazepines/quinoxaline. AB - Diversity-oriented synthesis of novel benzimidazole linked indolo benzodiazepine/quinoxaline ring systems using poly(ethylene glycol) as soluble polymer support is described. Commercially available 4-fluoro-3-nitrobenzoic acid and indoline were utilized for the construction of these annulated biheterocyclic compound libraries having multiple privileged structures with three-point structural diversity. A reagent based diversification approach coupled with the Pictet-Spengler-type condensation was used to construct the tetracyclic indolo benzodiazepines/quinoxalines on substituted benzimidazoles. PMID- 21648410 TI - 1,3-Bis(thiophosphinoyl)indene: a unique and versatile scaffold for original polymetallic complexes. AB - The coordination of tridentate ligands featuring lateral coordination sites prone to acting as bridging ligands was explored with the aim of obtaining original polymetallic species in a straightforward and controlled manner. Accordingly, the 2-indenylidene chloropalladate [{Ind(Ph(2)P?S)(2)}PdCl](-) was found to behave as a kappa(2)-C,S bidentate ligand toward metal fragments, giving access to homo- and heteropolymetallic complexes. X-ray diffraction analyses reveal the presence of short metal-metal contacts in all of these complexes. Density functional theory calculations unambiguously substantiate that the metals engage in unusual d(8)...d(8) interactions with a quasi-perpendicular arrangement of their coordination planes. PMID- 21648411 TI - Biosynthetic chlorination of the piperazate residue in kutzneride biosynthesis by KthP. AB - Kutznerides 2 and 8 of the cyclic hexadepsipeptide family of antifungal natural products from the soil actinomycete Kutzneria sp. 744 contain two sets of chlorinated residues, a 6,7-dichlorohexahydropyrroloindole moiety derived from dichlorotryptophan and a 5-chloropiperazate moiety, as well as a methylcyclopropylglycine residue that may arise from isoleucine via a cryptic chlorination pathway. Previous studies identified KtzD, KtzQ, and KtzR as three halogenases in the kutzneride pathway but left no candidate for installing the C5 chlorine on piperazate. On the basis of analysis of the complete genome sequence of Kutzneria, we now identify a fourth halogenase in the pathway whose gene is separated from the defined kutzneride cluster by 12 open reading frames. KthP (kutzneride halogenase for piperazate) is a mononuclear nonheme iron halogenase that acts on the piperazyl ring tethered by a thioester linkage to the holo forms of thiolation domains. MS analysis of the protein-bound product confirmed chlorination of the piperazate framework from the (3S)- but not the (3R) piperazyl-S-pantetheinyl thiolation proteins. After thioesterase-mediated release, nuclear magnetic resonance was used to assign the free imino acid as (3S,5S)-5-chloropiperazate, distinct from the 3S,5R stereoisomer reported in the mature kutznerides. These results demonstrate that a fourth halogenase, KthP, is active in the kutzneride biosynthetic pathway and suggest further processing of the (3S,5S)-5-chloropiperazate during subsequent incorporation into the kutzneride depsipeptide frameworks. PMID- 21648412 TI - Kinetics and mechanism of the oxidation of amaranth with hypochlorite. AB - The reaction mechanism of the oxidation of Amaranth dye (2-hydroxy-1-(4-sulfonato 1-naphthylazo) naphthalene-3,6-disulfonate) with hypochlorite under varied pH conditions was elucidated by a kinetic approach. Under excess concentration of oxidant, the reaction followed pseudo-first-order kinetics with respect to Amaranth, and the oxidation was found to occur through two competitive reactions, initiated by hypochlorite and hypochlorous acid. The reaction order with respect to both OCl(-) ion and HOCl was unity. While the latter reaction was fast, the significance of the oxidation paths depended on the relative concentration of the two oxidizing species, which was dictated by the reaction pH. The role of the H(+) ion in the reaction was established. For the hypochlorite ion and hypochlorous acid facilitated reactions, the second-order rate coefficients were 1.9 and 23.2 M(-1) s(-1), respectively. The energy parameters were E(a) = 33.7 kJ mol(-1), DeltaH(?) = 31.2 kJ mol(-1) and DeltaS(?) = -190.6 J K(-1) mol(-1) for the OCl(-) ion-driven oxidation, and E(a) = 26.9 kJ mol(-1), DeltaH(?) = 24.3 kJ mol(-1) and DeltaS(?) = -222.8 J K(-1) mol(-1) for the reaction with HOCl initiated oxidation. The major oxidation products for both the pathways were 3,4 dihydroxy naphthalene-2,7-disulfonic sodium salt (P(1)), dichloro-1,4 naphthoquione (P(2)) and naphtha(2,3)oxirene-2, 3-dione (P(3)). On the basis of the primary salt effect and other kinetic data, the rate law for the overall reaction and probable reaction mechanism was elucidated. The proposed mechanism was validated by simulations using Simkine-2. PMID- 21648413 TI - The vaporization enthalpies of 2- and 4-(N,N-dimethylamino)pyridine, 1,5 diazabicyclo[4.3.0]non-5-ene, 1,8-diazabicyclo[5.4.0]undec-7-ene, imidazo[1,2 a]pyridine and 1,2,4-triazolo[1,5-a]pyrimidine by correlation-gas chromatography. AB - The vaporization enthalpies of a series of heterocyclic compounds some suspected of showing evidence of a higher degree of self-association in comparison to many other nitrogen heterocycles have been measured both by correlation gas chromatography (CGC) and for comparison by transpiration. The compounds share some structural similarities. They include 2- and 4-(N,N-dimethylamino)pyridine, 1,5-diazabicyclo[4.3.0]non-5-ene, 1,8-diazabicyclo[5.4.0]undec-7-ene, 1,2,4 triazolo[1,5-a]pyrimidine and imidazo[1,2-a]pyridine. The vaporization enthalpies of the first three of the compounds are well reproduced by a standard series of reference compounds, mainly pyridine derivatives. The latter two compounds and 4 (N,N-dimethylamino)pyridine exhibit vaporization enthalpies approximately 7 kJ.mol(-1) larger. Their values are reproduced by using standards previously shown to exhibit stronger but similar self-association properties. The crystal structure of 1,2,4-triazolo[1,5-a]pyrimidine has been determined in an effort to understand the nature of the self-association. The molecule crystallizes in planar stacks slightly offset with a stacking distance of 3.24 A. PMID- 21648414 TI - Spontaneous dissociation of xenon tetroxide: phase and structural changes. AB - XeO(4) is a noble gas compound remarkable for its high explosiveness in the crystalline state and spontaneous explosion at melting temperature. Both phenomena are studied by analyzing potential energy surfaces corresponding to elementary dissociation acts. It is shown that a spontaneous explosion of xenon tetroxide can be explained by a phase transition associated with structural T(d) > D(2h) change and be triggered by rearrangement of electron levels due to the Jahn-Teller effect. PMID- 21648415 TI - Mitochondria are the main target organelle for trivalent monomethylarsonous acid (MMA(III))-induced cytotoxicity. AB - Excessive generation of reactive oxygen species (ROS) is considered to play an important role in arsenic-induced carcinogenicity in the liver, lungs, and urinary bladder. However, little is known about the mechanism of ROS-based carcinogenicity, including where the ROS are generated, and which arsenic species are the most effective ROS inducers. In order to better understand the mechanism of arsenic toxicity, rat liver RLC-16 cells were exposed to arsenite (iAs(III)) and its intermediate metabolites [i.e., monomethylarsonous acid (MMA(III)) and dimethylarsinous acid (DMA(III))]. MMA(III) (IC(50) = 1 MUM) was found to be the most toxic form, followed by DMA(III) (IC(50) = 2 MUM) and iAs(III) (IC(50) = 18 MUM). Following exposure to MMA(III), ROS were found to be generated primarily in the mitochondria. DMA(III) exposure resulted in ROS generation in other organelles, while no ROS generation was seen following exposures to low levels of iAs(III). This suggests the mechanisms of induction of ROS are different among the three arsenicals. The effects of iAs(III), MMA(III), and DMA(III) on activities of complexes I-IV in the electron transport chain (ETC) of rat liver submitochondrial particles and on the stimulation of ROS production in intact mitochondria were also studied. Activities of complexes II and IV were significantly inhibited by MMA(III), but only the activity of complexes II was inhibited by DMA(III). Incubation with iAs(III) had no inhibitory effects on any of the four complexes. Generation of ROS in intact mitochondria was significantly increased following incubation with MMA(III), while low levels of ROS generation were observed following incubation with DMA(III). ROS was not produced in mitochondria following exposure to iAs(III). The mechanism underlying cell death is different among As(III), MMA(III), and DMA(III), with mitochondria being one of the primary target organelles for MMA(III)-induced cytotoxicity. PMID- 21648416 TI - Open-shell singlet character of stable derivatives of nonacene, hexacene and teranthene. AB - The electronic ground states of the recently synthesized stable nonacene derivatives (J. Am. Chem. Soc. 2010, 132, 1261) are open-shell singlets with a polyradical nature instead of closed-shell singlets as originally assumed, according to the unrestricted broken spin-symmetry density functional theory (UBS DFT) computations (at B3LYP/6-31G*). It is the bulky protecting groups, not the transfer from the open-shell singlet to closed-shell singlet ground state, that stabilizes these longest characterized acenes. Similar analyses also confirmed the open-shell singlet character of the hexacene and teranthene derivatives. PMID- 21648417 TI - Efficient and regioselective 9-endo cyclization of alpha-carbamoyl radicals. AB - With the promotion of Lewis acid BF(3)*OEt(2), various N-(hex-5-enyl)-2 iodoalkanamides underwent efficient and regioselective 9-endo iodine-atom transfer radical cyclization reactions at room temperature. The cyclized products were readily converted to the corresponding azonan-2-ones by reduction with Bu(3)SnH or to hexahydroindolizin-3(5H)-ones by treatment with aqueous Na(2)CO(3) in a one-pot, two-stage manner. PMID- 21648418 TI - Unprecedented robust antiferromagnetism in fluorinated hexagonal perovskites. AB - The diversification of antiferromagnetic (AFM) oxides with high Neel temperature is of fundamental as well as technical interest if one considers the need for robust AFM in the field of spin-tronics (exchange bias, multiferroics, etc.). Within the broad series of so-called hexagonal perovskites (HP), the existence of face-sharing octahedral units drastically lowers the strength of magnetic exchanges as compared to corner-sharing octahedral edifices. Here, we show that the partial introduction of F(-) in several Fe-based HP types leads to a drastic increase of the AFM ordering close to the highest values reported in iron oxides (T(N) ~ 700 K). Our experimental results are supported by ab initio calculations. The T(N) increase is explained by the structural effect of the aliovalent F(-) for O(2-) substitution occurring in preferred anionic positions: it leads to local changes of the Fe-O-Fe connectivity and to chemical reduction into predominant Fe(3+), both responsible for drastic magnetic changes. PMID- 21648419 TI - Scalable fabrication of self-aligned graphene transistors and circuits on glass. AB - Graphene transistors are of considerable interest for radio frequency (rf) applications. High-frequency graphene transistors with the intrinsic cutoff frequency up to 300 GHz have been demonstrated. However, the graphene transistors reported to date only exhibit a limited extrinsic cutoff frequency up to about 10 GHz, and functional graphene circuits demonstrated so far can merely operate in the tens of megahertz regime, far from the potential the graphene transistors could offer. Here we report a scalable approach to fabricate self-aligned graphene transistors with the extrinsic cutoff frequency exceeding 50 GHz and graphene circuits that can operate in the 1-10 GHz regime. The devices are fabricated on a glass substrate through a self-aligned process by using chemical vapor deposition (CVD) grown graphene and a dielectrophoretic assembled nanowire gate array. The self-aligned process allows the achievement of unprecedented performance in CVD graphene transistors with a highest transconductance of 0.36 mS/MUm. The use of an insulating substrate minimizes the parasitic capacitance and has therefore enabled graphene transistors with a record-high extrinsic cutoff frequency (> 50 GHz) achieved to date. The excellent extrinsic cutoff frequency readily allows configuring the graphene transistors into frequency doubling or mixing circuits functioning in the 1-10 GHz regime, a significant advancement over previous reports (~20 MHz). The studies open a pathway to scalable fabrication of high-speed graphene transistors and functional circuits and represent a significant step forward to graphene based radio frequency devices. PMID- 21648420 TI - Comparison of lysophospholipid levels in rat feces with those in a standard chow. AB - Although lysophospholipids have attracted much attention due to their diverse physiological activities through their specific receptors, little is known about their metabolic fates in mammalian digestive systems after their ingestion as a minor food component. In this study, we analyzed five lysophospholipids in lipid extracts of a standard rat chow and feces of rats fed the chow by two-dimensional thin layer chromatography and liquid chromatography-tandem mass spectrometry. The most abundant lysophospholipid in the rat chow was lysophosphatidylcholine followed by lysophosphatidylethanolamine, lysophosphatidic acid (LPA), lysophosphatidylinositol and lysophosphatidylserine (LPS) in an increasing order, but their concentrations were very low in rat feces. Among the molecular species of LPS in the chow, only saturated species were detected in the feces in significant amounts. In addition, several molecular species of LPA remained in the feces in variable portions (saturated > monounsaturated > polyunsaturated). These results suggest that a portion of ingested LPA and LPS reach the rat large intestine, affecting physiological colon functions. PMID- 21648421 TI - Gene expression profiles of yeast Saccharomyces cerevisiae sod1 caused by patulin toxicity and evaluation of recovery potential of ascorbic acid. AB - Patulin (PAT) is a fungal secondary metabolite and exhibits various toxicities including DNA damage and oxidative stress. These toxicities are eased by ascorbic acid (AsA). Although a number of studies regarding the mitigating effect of AsA against PAT toxicity have been reported, a comprehensive study about gene expressions is currently underway. Here, we carried out a detailed evaluation of PAT toxicity by co-incubation with AsA using the superoxide dismutase (SOD) mutant. DNA microarray results extracted the alterations in iron transporter and Fe/S cluster assembly genes; some of the genes that constitute the cellular iron transporter systems remained dysfunctional even in the presence of AsA. Meanwhile, AsA treatment reduced the alterations of G1/S phase cell cycle regulation genes. These results suggest that oxidative stress-derived DNA damage still exists, although AsA treatment effectively reduces PAT toxicity. This implies that a combined condition is required for complete blockade of PAT toxicity. PMID- 21648422 TI - Electronic energy migration in solid versus liquid host matrices for concentrated perylenediimide dye solutions. AB - In this paper, we continue our evaluation of Forster-type theories of exciton diffusion in disordered environments. The perylenediimide dye Lumogen Red is used as a donor molecule in two different liquids, CHCl(3) and dimethylformamide, and the energy transfer to the acceptor molecule Rhodamine 700 is measured using time resolved fluorescence decays. The exciton motion is measured over Lumogen Red concentrations ranging from 1 * 10(-4) to 5 * 10(-2) M, and the results are compared to previous results for exciton diffusion in a solid polymer. Depending on the theoretical approach used to analyze the data, we find that the energy migration in the liquids is a factor of 2-3 faster than in the solid polymer, even after taking molecular translation into account. Measurements for a Lumogen Red concentration of 10 mM in the different host environments yield diffusion constants ranging from 2.2 to 3.1 nm(2)/ns in the liquids, as compared to 1.1-1.2 nm(2)/ns in solid poly(methyl methacrylate) (PMMA). The results in the liquids are in good agreement with theoretical predictions and numerical simulations of previous workers, while the results in solid PMMA are 2-3 times slower. This discrepancy is discussed in the context of the rapid energetic averaging present in the liquid environments but absent in the solid matrix, where unfavorable configurations and low energy trapping sites are frozen in by the static disorder. PMID- 21648423 TI - Exploring the sources of the magnetic anisotropy in a family of cyanide-bridged Ni9Mo6 and Ni9W6 systems: a density functional theory study. AB - A density functional theory (DFT) study of the magnetic coupling interactions and magnetic anisotropy in a family of experimentally synthesized Ni(9)Mo(V) and Ni(9)W(V) systems is presented. Our calculations show that for all of our selected Ni(9)M(6) systems, the intramolecular magnetic coupling interactions are ferromagnetic, and the ground-state spins are 12. All of the D values of Ni(9)W(6) systems come mainly from the contribution of the D(i) of W(6)(CN)(48)Ni extracted from Ni(9)W(6), and the influence of the eight surrounding Ni including the ligands on their magnetic anisotropy is very small. Although the surrounding Ni bounded by different ligands have a small influence on all D values for our selected complexes, they decide on the core structures of W(6)(CN)(48)Ni, which dominate their magnetic anisotropy. Thus, to obtain a Ni(9)W(6) system having a large negative D, we can use different ligands bound to Ni to obtain a good core structure of W(6)(CN)(48)Ni with a large negative D value. All D values of Ni(9)Mo(6) systems also come mainly from the contribution of D(i) of the Mo(6)(CN)(48)Ni, which is positive or negative but very small; most of these systems do not behave as single-molecule magnets. PMID- 21648424 TI - Surfactant self-assembly nanostructures in protic ionic liquids. AB - The existence and properties of mesoscopic self-assembly structures formed by surfactants in protic ionic liquid solutions are reported. Micellar aggregates of n-alkyltrimethylammonium (n = 10, 12, 14, 16) chlorides and bromides and of n alkylpyridinium (n = 12, 16) chlorides in ethylammonium nitrate and propylammonium nitrate were observed by means of several experimental techniques, including surface tension, transmission electron micrography, dynamic light scattering, and potentiometry using surfactant-selective electrodes. The effect of the alkyl chain length of both solute and solvent molecules on the critical micelle concentration is discussed, and a Stauff-Klevens law is seen to apply to surfactant solutions in both protic ionic liquids. The counterion role is also a matter of study in the case of alkyltrimethylammonium-based surfactants, and the presently reported evidence suggests that the place of the surfactant counterion in the Hoffmeister's series could determine its effect on micellization in IL solution. The size distribution of the aggregates is also analyzed together with the Gibbs free energies of micellization and the minimum surface area per monomer in all of the studied cases. All of the hereby reported evidence suggests that the negative entropic contribution arising from the release of the solvent layer upon micellization is also the driving force of conventional surfactant self association in protic ionic liquids. PMID- 21648425 TI - Modeling of chemical reactions in micelle: water-mediated keto-enol interconversion as a case study. AB - The effect of a zwitterionic micelle environment on the efficiency of the keto enol interconversion of 2-phenylacetylthiophene has been investigated by means of a joint application of experimental and theoretical/computational approaches. Results have revealed a reduction of the reaction rate constant if compared with bulk water essentially because of the different solvation conditions experienced by the reactant species, including water molecules, in the micelle environment. The slight inhibiting effect due to the application of a static electric field has also been theoretically investigated and presented. PMID- 21648426 TI - Regiodivergent annulation of alkynyl indoles to construct spiro-pseudoindoxyl and tetrahydro-beta-carbolines. AB - Regiodivergent annulations of 3-phenoxy alkynyl indoles have been developed and tuned by protective groups through gold catalysis. With electron-donating protective groups, the substrate followed a C3-selective annulation and gave structurally interesting tetrahydro-beta-carboline derivatives possessing potential bioactivity. Using electron-withdrawing protective groups, the substrate underwent a C2-selective annulation and afforded the structurally useful spiro-pseudoindoxyl found in natural indole alkaloids. Notably, an interesting and unusual 1, 2-migration of the phenoxy group was found in the C3 selective process. PMID- 21648427 TI - MicroRNA delivery by cationic lipoplexes for lung cancer therapy. AB - Lung cancer is the leading cause of cancer deaths in western countries and carries a poor overall five year survival rate. Several studies demonstrate that microRNAs (miRNAs or miRs) are actively involved in tumor development by serving as tumor suppressors, oncogenes or both. In lung cancer, miRNAs may serve as both diagnostic and prognostic biomarkers as well as regulate in vitro and in vivo tumor progression. However, miRNA-based therapy is faced with several challenges including lack of tissue specificity, lack of optimal delivery systems, poor cellular uptake and risk of systemic toxicity. Here, we report a cationic lipid based miRNA delivery system to address some of these challenges. Among many lung cancer related miRNAs, miR-133b, a tumor suppressor, was selected as a therapeutic target because it directly targets the prosurvival gene MCL-1 thus regulating cell survival and sensitivity of lung cancer cells to chemotherapeutic agents. The efficacy of pre-miR-133b containing lipoplexes was evaluated in A549 non-small cell lung cancer (NSCLC) cells. Compared with siPORT NeoFX transfection agent, lipoplexes delivered pre-miR-133b in a more efficient manner with ~2.3 fold increase in mature miR-133b expression and ~1.8-fold difference in MCL-1 protein downregulation in vitro. In the in vivo biodistribution study, lipoplexes achieved ~30% accumulation in lung tissue, which was ~50-fold higher than siPORT NeoFX transfection agent. Mice treated with pre-miR-133b containing lipoplexes had mature miR-133b expression in lung ~52-fold higher than untreated mice. Our results demonstrated that cationic lipoplexes are a promising carrier system for the development of miRNA-based therapeutics in lung cancer treatment. PMID- 21648428 TI - A chiral perazamacrocyclic fluorescent sensor for cascade recognition of Cu(II) and the unmodified alpha-amino acids in protic solutions. AB - A novel chiral Perazamacrocyclic fluorescent sensor (1) was designed and synthesized. It can serve as a fluorescent turn-off sensor with high selectivity toward Cu(II) among 14 metal ions. Furthermore, though 1 exhibits no enantioselectivity, after adding Cu(II), the in situ generated Cu(II)-containing complex of 1 (Cu(II)-1) can exhibit remarkable fluorescent enhancement responses and considerable enantioselectivities toward unmodified alpha-amino acids in protic solutions via a ligand displacement mechanism; i.e. a cascade recognition of Cu(II) and unmodified alpha-amino acids has been achieved. PMID- 21648429 TI - Nanoscale metal-organic frameworks for biomedical imaging and drug delivery. AB - Metal-organic frameworks (MOFs), a class of hybrid materials formed by the self assembly of polydentate bridging ligands and metal-connecting points, have been studied for a variety of applications. Recently, these materials have been scaled down to nanometer sizes, and this Account details the development of nanoscale metal-organic frameworks (NMOFs) for biomedical applications. NMOFs possess several potential advantages over conventional nanomedicines such as their structural and chemical diversity, their high loading capacity, and their intrinsic biodegradability. Under relatively mild conditions, NMOFs can be obtained as either crystalline or amorphous materials. The particle composition, size, and morphology can be easily tuned to optimize the final particle properties. Researchers have employed two general strategies to deliver active agents using NMOFs: by incorporating active agents into the frameworks or by loading active agents into the pores and channels of the NMOFs. The modification of NMOF surfaces with either silica coatings or organic polymers improves NMOF stability, fine-tunes their properties, and imparts additional functionality. Preliminary biomedical applications of NMOFs have focused on their use as delivery vehicles for imaging contrast agents and molecular therapeutics. Because NMOFs can carry large amounts of paramagnetic metal ions, they have been extensively explored as magnetic resonance imaging (MRI) contrast agents. Both Gd(3+)- and Mn(2+)-containing NMOFs have shown excellent efficacy as T(1) weighted contrast agents with large per metal- and per particle-based MR relaxivities. Fe(3+)-containing NMOFs have demonstrated excellent T(2)-weighted contrast enhancement. Upon intravenous injection of iron carboxylate NMOFs in Wistar rats, researchers observed negative signal enhancement in the liver and spleen, which dissipated over time, indicating the degradation and clearance of the NMOF. Through the incorporation of luminescent or high Z element building blocks, NMOFs have also served as viable contrast agents for optical imaging or X ray computed tomography (CT) imaging. Incorporation of membrane impermeable dyes into NMOFs allowed for their uptake by cancer cells and for their controlled release as the framework decomposed. NMOFs have been used to deliver anticancer drugs and other chemotherapeutics. Cisplatin prodrugs were incorporated within NMOFs at exceptionally high levels, either through use of the prodrug as the building block or through attachment of the prodrug onto the framework after synthesis. These NMOFs were encapsulated within a silica shell and targeted to cancer cells. In vitro assays revealed that the targeted NMOFs possessed similar efficacy to cisplatin, while the nontargeted NMOFs were less active. Several different therapeutic molecules were loaded within porous iron-carboxylate NMOFs at unprecedented levels. The NMOF showed sustained drug release with no burst effect, and in vitro assays revealed that the nanoencapsulated drug possessed similar efficacy to the free drug. Although still at a very early stage of development, NMOFs have already shown great promise as a novel platform for nanomedicine. The compositional tunability and mild synthetic conditions used to produce NMOFs should allow for the incorporation of other imaging and therapeutic agents and their effective delivery to targeted cells in vivo. PMID- 21648430 TI - Functionalization of inorganic nanoparticles for bioimaging applications. AB - Modern biomedical imaging technologies have led to significant advances in diagnosis and therapy. Because most disease processes occur at the molecular and cellular levels, researchers continue to face challenges in viewing and understanding these processes precisely and in real time. The ideal imaging resolution would be in nanometers, because most biological processes take place on this length scale. Therefore, the functionalization of nanoparticles (NPs) and their use in therapeutic and diagnostic applications are of great interest. Molecular and cellular imaging agents made from inorganic NPs have been developed to probe such biological events noninvasively. The conjugation of tiny NPs with specific biomolecules allows researchers to target the desired location, reduce overall toxicity, and boost the efficiency of the imaging probes. In this Account, we review recent research on the functionalization of NPs for bioimaging applications. Several types of NPs have been employed for bioimaging applications, including metal (Au, Ag), metal oxide (Fe(3)O(4)), and semiconductor nanocrystals (e.g. quantum dots (QDs) and magnetic quantum dots (MQDs)). The preparation of NPs for bioimaging applications can include a variety of steps: synthesis, coating, surface functionalization, and bioconjugation. The most common strategies of engineering NP surfaces involve physical adsorption or chemisorption of the desired ligands onto the surface. Chemisorption or covalent linkages are preferred, and the coated NPs should possess high colloidal stability, biocompatibility, water solubility, and functional groups for further bioconjugation. Many of the functionalization techniques that have been reported in the literature suffer from limitations such as complex synthesis steps, poor biocompatibility, low stability, and hydrophobic products. Coating strategies based on chemisorption and ligand exchange often provide a better way to tailor the surface properties of NPs. After conjugation with the appropriate targeting ligands, antibodies, or proteins, the NPs may exhibit highly selective binding, making them useful for fluorescence imaging, magnetic resonance imaging (MRI), positron emission tomography (PET) imaging, and multimodal imaging. PMID- 21648431 TI - Redox and pH microenvironments within Shewanella oneidensis MR-1 biofilms reveal an electron transfer mechanism. AB - The goal of this research was to quantify the variations in redox potential and pH in Shewanella oneidensis MR-1 biofilms respiring on electrodes. We grew S. oneidensis MR-1 on a graphite electrode, which was used to accept electrons for microbial respiration. We modified well-known redox and pH microelectrodes with a built-in reference electrode so that they could operate near polarized surfaces and quantified the redox potential and pH profiles in these biofilms. In addition, we used a ferri-/ferrocyanide redox system in which electrons were only transferred by mediated electron transfer to explain the observed redox potential profiles in biofilms. We found that regardless of the polarization potential of the biofilm electrode, the redox potential decreased toward the bottom of the biofilm. In a fully redox-mediated control system (ferri-/ferrocyanide redox system), the redox potential increased toward the bottom when the electrode was the electron acceptor. The opposite behavior of redox profiles in biofilms and the redox-controlled system is explained by S. oneidensis MR-1 biofilms not being redox-controlled when they respire on electrodes. The lack of a significant variation in pH implies that there is no proton transfer limitation in S. oneidensis MR-1 biofilms and that redox potential profiles are not caused by pH. PMID- 21648433 TI - Highly linear self-assembled porphyrin wires. AB - An efficient noncovalent assembly process involving high geometrical control was applied to a linear bis(imidazolyl zinc porphyrin) 7Zn, bearing C(18) substitutents, to generate linear multiporphyrin wires. The association process is based on imidazole recognition within the cavity of the phenanthroline strapped zinc porphyrin. In chlorinated solvents, discrete soluble oligomers were obtained after (7Zn)(n) was end-capped with a terminal single imidazolyl zinc porphyrin derivative 4Zn. These soluble species, as well as their destabilization in the presence of protic solvents, were studied by UV-visible and time-resolved luminescence. In the solid state, assemblies as long as 480 nm, which corresponds to 190 iterative units or a total of 380 porphyrins, were observed by atomic force microscopy measurements on mica. The length and linearity of the porphyrin wires obtained illustrate the potential of phenanthroline-strapped porphyrins for the directional control of self-assembly processes. PMID- 21648432 TI - Structure of the HIV-1 frameshift site RNA bound to a small molecule inhibitor of viral replication. AB - Programmed -1 translational frameshifting is an essential event in the replication cycle of HIV. Frameshifting is required for expression of the viral Pol proteins, and drug-like molecules that target this process may inhibit HIV replication. A small molecule stimulator of HIV-1 frameshifting and inhibitor of viral replication, DB213 (RG501), was previously discovered from a high throughput screen. However, the mechanistic basis for this compound's effects was unknown, and to date no structural information exists for small molecule effectors of frameshifting. Here, we investigate the binding of DB213 to the frameshift site RNA and have determined the structure of this complex by NMR. Binding of DB213 stabilizes the RNA and increases its melting temperature by 10 degrees C. The ligand binds to a primary site on the RNA stem-loop, although nonspecific interactions are also detected. The compound binds in the major groove and spans a distance of 9 base pairs. DB213 hydrogen bonds to phosphate groups on opposite sides of the major groove and alters the conformation of a conserved GGA bulge in the RNA. This study may provide a starting point for structure-based optimization of compounds targeting the HIV-1 frameshift site RNA. PMID- 21648434 TI - Nickel phase partitioning and toxicity in field-deployed sediments. AB - The pool of bioavailable metal in sediments can be much smaller than total metal concentration due to complexation and precipitation with ligands. Metal bioavailability and toxicity in sediment is often predicted from models of simultaneous extracted metal and acid volatile sulfide (SEM-AVS); however, studies of the applicability of these models for Ni-contaminated sediments have been conducted primarily in laboratory settings. We investigated the utility of the SEM-AVS models under field conditions: Five lotic sediments with a range of sulfide and organic carbon contents were amended with four concentrations of Ni, deployed in streams for eight weeks, and examined for colonizing macroinvertebrates. After four weeks, colonizing macroinvertebrates showed a strong negative response to the Ni-treated sediments and SEM-AVS models of bioavailability differentiated between toxic and nontoxic conditions. By Week 8, relationships deteriorated between colonizing macroinvertebrates and SEM-AVS model predictions. Total Ni in the sediment did not change through time; however, Ni partitioning shifted from being dominated by organic cabon at deployment to associations with Fe and Mn. Combined geochemical and toxicity results suggest that Fe and Mn oxides in surface sediments resulted in Ni being less available to biota. This implies that current SEM-AVS models may overestimate bioavailable Ni in sediments with oxic surface layers and sufficient Fe and Mn. PMID- 21648435 TI - In situ calibration of a passive sampling device for selected illicit drugs and their metabolites in wastewater, and subsequent year-long assessment of community drug usage. AB - Polar organic chemical integrative samplers (POCIS) were calibrated in situ for selected illicit drugs and their metabolites at a sewage treatment works. Eleven out of 13 target compounds were detected and eight of those exhibited linear uptake kinetics with sampling rates between 0.035 and 0.150 L d(-1). Subsequently POCIS were deployed for 2 week periods over the course of a whole year, in order to examine trends in drug usage. Amphetamine and methamphetamine showed several similar peaks in concentration during the course of the year as did cocaine and two of its metabolites. Low levels of ecstasy were observed, with a prominent peak in May and a steady increase toward the end of the year. The antihistamine Cetirizine showed a clear increase in use during the summer months as expected and back calculation of the yearly dosage from POCIS accumulations yielded very similar results to that registered in the Norwegian prescription database. Estimations of cocaine usage using the parent compound averaged between 0.31 and 2.8 g d(-1) per 1000 inhabitants. POCIS is a cost-effective technique for the long-term monitoring of drug usage of a defined population and may overcome the difficulties of representative sampling associated with autosampling equipment. PMID- 21648436 TI - Competitive microbially and Mn oxide mediated redox processes controlling arsenic speciation and partitioning. AB - The speciation and partitioning of arsenic (As) in surface and subsurface environments are controlled, in part, by redox processes. Within soils and sediments, redox gradients resulting from mass transfer limitations lead to competitive reduction-oxidation reactions that drive the fate of As. Accordingly, the objective of this study was to determine the fate and redox cycling of As at the interface of birnessite (a strong oxidant in soil with a nominal formula of MnO(x), where x ~ 2) and dissimilatory As(V)-reducing bacteria (strong reductant). Here, we investigate As reduction-oxidation dynamics in a diffusively controlled system using a Donnan reactor where birnessite and Shewanella sp. ANA 3 are isolated by a semipermeable membrane through which As migrates. Arsenic(III) injected into the reaction cell containing birnessite is rapidly oxidized to As(V). Arsenic(V) diffusing into the Shewanella chamber is then reduced to As(III), which subsequently diffuses back to the birnessite chamber, undergoing oxidation, and establishing a continuous cycling of As. However, we observe a rapid decline in the rate of As(III) oxidation owing to passivation of the birnessite surface. Modeling and experimental results show that high [Mn(II)] combined with increasing [CO(3)(2-)] from microbial respiration leads to the precipitation of rhodochrosite, which eventually passivates the Mn oxide surface, inhibiting further As(III) oxidation. Our results show that despite the initial capacity of birnessite to rapidly oxidize As(III), the synergistic effect of intense As(V) reduction by microorganisms and the buildup of reactive metabolites capable of passivating reactive mineral surfaces-here, birnessite-will produce (bio)geochemical conditions outside of those based on thermodynamic predictions. PMID- 21648437 TI - Haloacetic acids in swimming pools: swimmer and worker exposure. AB - For the first time, the exposure of swimmers and workers to haloacetic acids (HAAs) in indoor and outdoor pools was evaluated through the analysis of urine samples. The subjects of this study, 49 volunteers, were male and female workers as well as swimmers (adults and children) who regularly attended an indoor pool (January-June) and an outdoor one (July and August). The results showed that HAAs appeared 20-30 min after exposure and were eliminated within 3 h. After 2 h exposure, urine samples taken from workers contained dichloroacetic (DCAA) and trichloroacetic (TCAA) acids at ~300 and ~120 ng/L levels since HAAs were aerosolized in the indoor ambient, whereas only DCAA was found in some workers' urine samples from the outdoor pool but at ~50 ng/L levels, despite the fact that the outdoor pools generally had somewhat higher levels of HAAs than the indoor pools. After 1 h swimming TCAA, DCAA and MCAA were present at concentrations of ~4400, ~2300, and ~560 ng/L, respectively, in the swimmers' urine in the indoor pool; similar results were obtained from the swimmers in the outdoor pool due to accidental ingestion. Finally, exposure estimates indicate that ingestion is the major route of exposure (~94%), followed by inhalation (~5%) and dermal contribution (~1%). PMID- 21648439 TI - Lattice vibrational modes and their frequency shifts in semiconductor nanowires. AB - We have performed first-principles calculations to study the lattice vibrational modes and their Raman activities in silicon nanowires (SiNWs). Two types of characteristic vibrational modes are examined: high-frequency optical modes and low-frequency confined modes. Their frequencies have opposite size dependence with a red shift for the optical modes and a blue shift for the confined modes as the diameter of SiNWs decreases. In addition, our calculations show that these vibrational modes can be detected by Raman scattering measurements, providing an efficient way to estimate the size of SiNWs. PMID- 21648438 TI - Time-dependent fifth-order bands in nominally third-order 2D IR vibrational echo spectra. AB - Progress in the field of 2D IR vibrational spectroscopy has been bolstered by the production of intense mid-IR laser pulses. As higher-energy pulses are employed, a concomitant increase occurs in the likelihood of fifth-order contributions to the 2D IR spectra. We report the appearance of fifth-order signals in 2D IR spectra of CO bound to the active site of the enzyme cytochrome P450(cam) with the substrate norcamphor. Two bands with novel time dependences, one on the diagonal and one off-diagonal, are not accounted for by normal third-order interactions. These bands are associated with a nu = 1-2 vibrational transition frequency. Both bands decay to 0 and then grow back in with opposite sign. The diagonal band is positive at short time, decays to 0, reappears with negative sign, before eventually decaying to 0. The off-diagonal band is negative at short time, decays to 0, reappears positive, and then decays to 0. The appearance and time dependence of these bands are characterized. Understanding these fifth-order bands is useful because they may be misidentified with time-dependent bands that arise from other processes, such as chemical exchange, vibrational coupling, or energy transfer. The presence and unusual time dependences of the fifth-order bands are reproduced with model calculations that account for the fact that vibrational relaxation from the nu = 2 to 1 level is approximately a factor of 2 faster than that from the nu = 1 to 0 level. PMID- 21648440 TI - Effects of extending the computational model on DNA-protein T-shaped interactions: the case of adenine-histidine dimers. AB - The MP2/6-31G*(0.25) pi-pi or pi(+)-pi T-shaped (edge-to-face) interactions between neutral or protonated histidine and adenine were considered using computational models of varying size to determine the effects of the protein and DNA backbones on the preferred dimer structure and binding strength. The overall consequences of the backbones are reasonably subtle for the neutral adenine histidine T-shaped dimers. Furthermore, the minor changes in the binding strengths of these dimers upon model extension arise from additional (attractive) backbone-pi (bb-pi) contacts and changes in the preferred pi-pi orientations, which is verified by the quantum theory of atoms in molecules (QTAIM). Since the binding strength of the extended dimer equals the sum of the individual backbone pi and pi-pi contributions, the pi-pi component is not appreciably affected by polarization of the ring upon inclusion of the biological backbone. In contrast, the larger effect of the backbone on the protonated histidine dimers cannot simply be predicted as the sum of changes in the pi-pi and bb-pi components regardless of the dimer type or model. This suggests, and QTAIM qualitatively supports, that the magnitude of the pi(+)-pi contribution changes, which is likely due to alterations in the electrostatic landscape of the monomer rings upon inclusion of the biological backbone that largely affect T-shaped dimers. These findings differ from those previously reported for (neutral) pi-pi stacked and (metallic) cation-pi interactions, which highlights the distinct properties of each (pi-pi, pi(+)-pi, and cation-pi) classification of noncovalent interaction. Furthermore, these results emphasize the importance of considering backbone-pi interactions when analyzing contacts that appear in experimental crystal structures and cautions the use of truncated models when evaluating the magnitude of the pi(+)-pi contribution present in large biological complexes. PMID- 21648441 TI - Absorption properties of metal-semiconductor hybrid nanoparticles. AB - The optical response of hybrid metal-semiconductor nanoparticles exhibits different behaviors due to the proximity between the disparate materials. For some hybrid systems, such as CdS-Au matchstick-shaped hybrids, the particles essentially retain the optical properties of their original components, with minor changes. Other systems, such as CdSe-Au dumbbell-shaped nanoparticles, exhibit significant change in the optical properties due to strong coupling between the two materials. Here, we study the absorption of these hybrids by comparing experimental results with simulations using the discrete dipole approximation method (DDA) employing dielectric functions of the bare components as inputs. For CdS-Au nanoparticles, the DDA simulation provides insights on the gold tip shape and its interface with the semiconductor, information that is difficult to acquire by experimental means alone. Furthermore, the qualitative agreement between DDA simulations and experimental data for CdS-Au implies that most effects influencing the absorption of this hybrid system are well described by local dielectric functions obtained separately for bare gold and CdS nanoparticles. For dumbbell shaped CdSe-Au, we find a shortcoming of the electrodynamic model, as it does not predict the "washing out" of the optical features of the semiconductor and the metal observed experimentally. The difference between experiment and theory is ascribed to strong interaction of the metal and semiconductor excitations, which spectrally overlap in the CdSe case. The present study exemplifies the employment of theoretical approaches used to describe the optical properties of semiconductors and metal nanoparticles, to achieve better understanding of the behavior of metal-semiconductor hybrid nanoparticles. PMID- 21648442 TI - Enhanced sensing of nonpolar volatile organic compounds by silicon nanowire field effect transistors. AB - Silicon nanowire field effect transistors (Si NW FETs) are emerging as powerful sensors for direct detection of biological and chemical species. However, the low sensitivity of the Si NW FET sensors toward nonpolar volatile organic compounds (VOCs) is problematic for many applications. In this study, we show that modifying Si NW FETs with a silane monolayer having a low fraction of Si-O-Si bonds between the adjacent molecules greatly enhances the sensitivity toward nonpolar VOCs. This can be explained in terms of an indirect sensor-VOC interaction, whereby the nonpolar VOC molecules induce conformational changes in the organic monolayer, affecting (i) the dielectric constant and/or effective dipole moment of the organic monolayer and/or (ii) the density of charged surface states at the SiO(2)/monolayer interface. In contrast, polar VOCs are sensed directly via VOC-induced changes in the Si NW charge carriers, most probably due to electrostatic interaction between the Si NW and polar VOCs. A semiempirical model for the VOC-induced conductivity changes in the Si NW FETs is presented and discussed. PMID- 21648444 TI - Calcium-decorated carbyne networks as hydrogen storage media. AB - Among the carbon allotropes, carbyne chains appear outstandingly accessible for sorption and very light. Hydrogen adsorption on calcium-decorated carbyne chain was studied using ab initio density functional calculations. The estimation of surface area of carbyne gives the value four times larger than that of graphene, which makes carbyne attractive as a storage scaffold medium. Furthermore, calculations show that a Ca-decorated carbyne can adsorb up to 6 H(2) molecules per Ca atom with a binding energy of ~0.2 eV, desirable for reversible storage, and the hydrogen storage capacity can exceed ~8 wt %. Unlike recently reported transition metal-decorated carbon nanostructures, which suffer from the metal clustering diminishing the storage capacity, the clustering of Ca atoms on carbyne is energetically unfavorable. Thermodynamics of adsorption of H(2) molecules on the Ca atom was also investigated using equilibrium grand partition function. PMID- 21648445 TI - A study on applications of N-substituted main-chain NHC-palladium polymers as recyclable self-supported catalysts for the Suzuki-Miyaura coupling of aryl chlorides in water. AB - The preparation and characterization of a number of main-chain organometallic polymers (NHC-Pd MCOP) with different N-alkyl substituted groups such as benzyl (3a), n-hexyl (3b), and n-dodecyl (3c) are described. Among these polymers, 3c bearing the more lipophilic group n-dodecyl was found to be a more reactive and recoverable catalytic system in the Suzuki-Miyaura cross-coupling reaction of chloroarenes, including both deactivated and hindered aryl chlorides with different types of arylboronic acids under aqueous conditions. While the catalysts seem to be highly recyclable, on the contrary, we have provided much compelling evidence, such as kinetic monitoring, poisoning experiments, and average molecular weight determination before and after catalysis, that shows that the described organometallic polymers might be indeed the source of production of active soluble Pd species in the form of either Pd nanoparticles or fragmented NHC-Pd complexes. Our studies showed that in order to assess whether the catalysts are functioning in a heterogeneous pathway or they are simply a source of production of active Pd species, it is crucial to devise a suitable and highly efficient poison that could capture essentially soluble catalytic species. In this regard, we interestingly found that among a variety of well-known catalyst poisons such as Hg(0), SBA-15-PrSH, and cross-linked poly(4 vinylpyridine) (PVP), only PVP could efficiently quench catalysis, thus providing clear evidence of the formation soluble Pd species in our protocol. In addition, several experiments such as bright-field microscopy, dynamic light scattering (DLS) of the reaction mixture, and kinetic monitoring of the reaction at an early stage confirm not only that the described organometallic polymers could be a source of production of trace amounts of Pd nanoparticles but the capsular structures of these lipophilic polymers in water provides a means of entrapment of nanoclusters in a hydrophobic region, thus accelerating the reaction in pure water in the absence of any co-organic solvent. PMID- 21648446 TI - Water molecule-induced stiffening in ZnO nanobelts. AB - We report the observation of remarkable water molecule-induced stiffening in ZnO nanobelts using atomic force microscopy three-point bending test. It was found that the elastic modulus of ZnO nanobelts could increase significantly from 40 GPa under ambient condition up to 88 GPa at the relative humidity level of 80%. The physical mechanism for this phenomenon was explained in terms of increasing surface stress induced by water molecule adsorption on ZnO nanobelt surface. Our first-principles density functional theory calculations revealed that the water molecules adsorbed on the ZnO surface would attract surface Zn atoms to move outward and hence increase the value of surface stress of ZnO surface. PMID- 21648447 TI - Component dynamics in miscible mixtures of water and methanol. AB - In binary mixtures with hydrophilic substances, water is usually the more mobile component and its relaxation time is shorter than those of the other components. An exception is the case of the mixture of 1-propanol with 45 mol % water, where the alpha-relaxation of water is slower than the alpha-relaxation of 1-propanol and even slower than the local relaxation of water confined in various spaces of nanometer size. This unusual result, so far obtained in a mixture of 1-propanol with water at a single composition, deserves confirmation by experiments in another mixture at more than one composition. Toward this goal, we have chosen mixtures of methanol with water at concentrations of water ranging from 10 to 40 mol % and investigated the dynamics of the slower water and the faster methanol components by broad-band dielectric relaxation measurements. The alpha-relaxation time of the water component becomes shorter with increasing content of the faster methanol component in the mixture as expected and is much shorter than in the mixture of 1-propanol with 45 mol % water. In mixtures with lower water contents of 10-20 mol %, the alpha-relaxation of the methanol component has a narrow frequency dispersion and no resolved Johari-Goldstein beta-relaxation, indicating a low degree of intermolecular coupling or cooperativity of methanol. An increase of the content of the slower water component effectively enhances intermolecular coupling of the methanol component. Consequently, the alpha-relaxation of the methanol component becomes more cooperative, as evidenced by broadening of its frequency dispersion and the appearance of a resolved Johari-Goldstein beta relaxation of methanol when the water concentration is higher than 30 mol %. The observations are rationalized by application of the coupling model. PMID- 21648448 TI - Surface interaction forces of cellulose nanocrystals grafted with thermoresponsive polymer brushes. AB - The colloidal stability and thermoresponsive behavior of poly(N isopropylacrylamide) brushes grafted from cellulose nanocrystals (CNCs) of varying graft densities and molecular weights was investigated. Indication of the grafted polymer brushes was obtained after AFM imaging of CNCs adsorbed on silica. Also, aggregation of the nanoparticles carrying grafts of high degree of polymerization was observed. The responsiveness of grafted CNCs in aqueous dispersions and as an ultrathin film was evaluated by using light scattering, viscosimetry, and colloidal probe microscopy (CPM). Light transmittance measurements showed temperature-dependent aggregation originating from the different graft densities and molecular weights. The lower critical solution temperature (LCST) of grafted poly(NiPAAm) brushes was found to decrease with the ionic strength, as is the case for free poly(NiPAAm) in aqueous solution. Thermal responsive behavior of grafted CNCs in aqueous dispersions was observed by a sharp increase in dispersion viscosity as the temperature approached the LCST. CPM in liquid media for asymmetric systems consisting of ultrathin films of CNCs and a colloidal silica probe showed the distinctive effects of the grafted polymer brushes on interaction and adhesive forces. The origin of such forces was found to be mainly electrostatic and steric in the case of bare and grafted CNCs, respectively. A decrease in the onset of attractive and adhesion forces of grafted CNCs films were observed with the ionic strength of the aqueous solution. The decreased mobility of polymer brushes upon partial collapse and decreased availability of hydrogen bonding sites with higher electrolyte concentration were hypothesized as the main reasons for the less prominent polymer bridging between interacting surfaces. PMID- 21648449 TI - The [MoFe3S4]2+ oxidation state: synthesis, substitution reactions, and structures of phosphine-ligated cubane-type clusters with the S=2 ground state. AB - The cluster [(Tp)MoFe(3)S(4)(PEt(3))(3)](1+) containing the cubane-type [MoFe(3)(MU(3)-S)(4)](2+) reduced core undergoes facile ligand substitution reactions at the iron sites leading to an extensive set of mono- and disubstituted species [(Tp)MoFe(3)S(4)(PEt(3))(3-n)L(n)](1-n) with L = halide, N(3)(-), PhS(-), PhSe(-), R(3)SiO(-), and R(3)SiS(-) and n = 1 and 2. Structures of 10 members of the set are reported. For two representative clusters, Curie behavior at 2-20 K indicates a spin-quintet ground state. Zero-field Mossbauer spectra consist of two doublets in a 2:1 intensity ratio. (57)Fe isomer shifts are consistent with the mean oxidation state Fe(3)(2.33+) arising from electron delocalization of the mixed-valence oxidation state description [Mo(3+)Fe(3+)Fe(2+)(2)]. Reaction of [(Tp)MoFe(3)S(4)(PEt(3))(2)Cl] with (Me(3)Si)(2)S affords [(Tp)MoFe(3)S(4)(PEt(3))(2)(SSiMe(3))], a likely first intermediate in the formation of the tricluster compound {[(Tp)MoFe(3)S(4)(PEt)(2)](3)S}(BPh(4)) from the reaction of [(Tp)MoFe(3)S(4)(PEt(3))(3)](BPh(4)) and NaSSiMe(3) in tetrahydrofuran (THF). The tricluster consists of three cluster units bound to a central MU(3)-S atom in a species of overall C(3) symmetry. Relatively few clusters in the [MoFe(3)S(4)](2+) oxidation state have been prepared compared to the abundance of clusters in the oxidized [MoFe(3)S(4)](3+) state. This work is the first comprehensive study of the [MoFe(3)S(4)](2+) state, one conspicuous feature of which is its ability to bind hard and soft sigma-donors and strong to weak pi acid ligands. (Tp = tris(pyrazolyl)hydroborate(1-)). PMID- 21648450 TI - Chemical reactivity and electrochemistry of metal-metal-bonded zincocenes. AB - Attempts to prepare mixed-ligand zinc-zinc-bonded compounds that contain bulky C(5)Me(5) and terphenyl groups, [Zn(2)(C(5)Me(5))(Ar')], lead to disproportionation. The resulting half-sandwich Zn(II) complexes [(eta(5) C(5)Me(5))ZnAr'] (Ar' = 2,6-(2,6-(i)Pr(2)C(6)H(3))(2)-C(6)H(3), 2; 2,6-(2,6 Me(2)C(6)H(3))(2)-C(6)H(3), 3) can also be obtained from the reaction of [Zn(C(5)Me(5))(2)] with the corresponding LiAr'. In the presence of pyr-py (4 pyrrolidinopyridine) or DBU (1,8-diazabicyclo[5.4.0]undec-7-ene), [Zn(2)(eta(5) C(5)Me(5))(2)] reacts with C(5)Me(5)OH to afford the tetrametallic complexes [Zn(2)(eta(5)-C(5)Me(5))L(MU-OC(5)Me(5))](2) (L = pyr-py, 6; DBU, 8), respectively. The bulkier terphenyloxide Ar(Mes)O(-) group (Ar(Mes) = 2,6-(2,4,6 Me(3)C(6)H(2))(2)-C(6)H(3)) gives instead the dimetallic compound [Zn(2)(eta(5) C(5)Me(5))(OAr(Mes))(pyr-py)(2)], 7, that features a terminal Zn-OAr(Mes) bond. DFT calculations on models of 6-8 and also on the Zn-Zn-bonded complexes [Zn(2)(eta(5)-C(5)H(5))(OC(5)H(5))(py)(2)] and [(eta(5)-C(5)H(5))ZnZn(py)(3)](+) have been performed and reveal the nonsymmetric nature of the Zn-Zn bond with lower charge and higher participation of the s orbital of the zinc atom coordinated to the cyclopentadienyl ligand with respect to the metal within the pseudo-ZnL(3) fragment. Cyclic voltammetric studies on [Zn(2)(eta(5) C(5)Me(5))(2)] have been also carried out and the results compared with the behavior of [Zn(C(5)Me(5))(2)] and related magnesium and calcium metallocenes. PMID- 21648451 TI - Investigating the quartz (1010)/water interface using classical and ab initio molecular dynamics. AB - Two different terminations of the (1010) surface of quartz (alpha and beta) interacting with water are simulated by classical (CMD) (using two different force fields) and ab initio molecular dynamics (AIMD) and compared with previously published X-ray reflectivity (XR) experiments. Radial distribution functions between hydroxyl and water show good agreement between AIMD and CMD using the ClayFF force field for both terminations. The Lopes et al. (Lopes, P. E. M.; Murashov, V.; Tazi, M.; Demchuk, E.; MacKerell, A. D. J. Phys. Chem. B2006, 110, 2782-2792) force field (LFF), however, underestimates the extent of hydroxyl-water hydrogen bonding. The beta termination is found to contain hydroxyl-hydroxyl hydrogen bonds; the quartz surface hydroxyl hydrogens and oxygens that hydrogen bond with each other exhibit greatly reduced hydrogen bonding to water. Conversely, the hydroxyl hydrogen and oxygens that are not hydrogen bonded to other surface hydroxyls but are connected to those that are show a considerable amount of hydrogen bonding to water. The electron density distribution of an annealed surface of quartz (1010) obtained by XR is in qualitative agreement with electron densities calculated by CMD and AIMD. In all simulation methods, the interfacial water peak appears farther from the surface than observed by XR. Agreement among AIMD, LFF, and XR is observed for the relaxation of the near-surface atoms; however, ClayFF shows a larger discrepancy. Overall, results show that for both terminations of (1010), LFF treats the near surface structure more accurately whereas ClayFF treats the interfacial water structure more accurately. It is shown that the number of hydroxyl and water hydrogen bonds to the bridging Si-O-Si oxygens connecting the surface silica groups to the rest of the crystal is much greater for the alpha than the beta termination. It is suggested that this may play a role in the greater resistance to dissolution of the beta termination than that of the alpha termination. PMID- 21648452 TI - Graphene spintronic devices with molecular nanomagnets. AB - The possibility to graft nano-objects directly on its surface makes graphene particularly appealing for device and sensing applications. Here we report the design and the realization of a novel device made by a graphene nanoconstriction decorated with TbPc(2) magnetic molecules (Pc = phthalocyananine), to electrically detect the magnetization reversal of the molecules in proximity with graphene. A magnetoconductivity signal as high as 20% is found for the spin reversal, revealing the uniaxial magnetic anisotropy of the TbPc(2) quantum magnets. These results depict the behavior of multiple-field-effect nanotransistors with sensitivity at the single-molecule level. PMID- 21648453 TI - beta-Olefination of 2-alkynoates leading to trisubstituted 1,3-dienes. AB - A phosphine-mediated olefination of 2-alkynoates with aldehydes forming 1,3 dienes with high E-selectivity and up to 88% yield is described. Reaction conditions are optimized and reactions are demonstrated for various aryl, alkyl, and alkenyl aldehydes and for ethyl 2-alkynoates with different substituents in the delta-position. Proof of concept is shown for the generation of a beta,gamma unsaturated lactone by intramolecular olefination, and furthermore the use of the generated 1,3-dienes in the Diels-Alder reaction has been demonstrated. PMID- 21648454 TI - Fusing triazoles: toward extending aromaticity. AB - A novel method to extend aromaticity by one benzene and two triazole rings was developed and optimized. This two-step route employs the copper-catalyzed azide haloalkyne cycloaddition reaction of an ortho-bis(iodoacetylene) system and the subsequent intramolecular homocoupling fusion of the neighboring iodotriazoles, a process in which an additional benzene ring is formed. This versatile methodology allows one to extend the core size of chromophores and, consequently, to tune the material's properties. PMID- 21648455 TI - Ring strain-promoted allylic transposition of cyclic silyl ethers. AB - Relief of the ring strain of medium-sized rings promotes a regioselective allylic transposition of a C-O bond when catalyzed by rhenium oxide. Through the allylic transposition, eight-membered cyclic silyl ethers undergo ring contraction to the corresponding six-membered siloxacycles. PMID- 21648456 TI - Synthesis, characterization, and photophysical study of fluorescent N-substituted benzo[ghi]perylene "swallow tail" monoimides. AB - A set of N-substituted benzoperylene monoimide (BPI) fluorophores was synthesized and characterized structurally and photophysically. Condensation of benzo[ghi]perylene-1,2-dicarboxylic anhydride in the presence of "swallow tail" alkyl amines produced fluorophores that are soluble in a range of organic solvents, highly absorbing in the near-UV (epsilon(334) = 79,000 M(-1) cm(-1)), and fluorescent in the visible range. Photophysical behavior of the compounds was studied with steady-state and time-correlated single photon counting. The synthesized BPIs exhibit positive solvachromatic emission (lambda(em) (hexane) = 469 nm; lambda(em) (ethanol) = 550 nm) as a function of solvent polarity with little change in their excited-state lifetime (9.6-6.5 ns) and fluorescence quantum yield (0.27-0.44) over the polarity range studied. Solvachromatic shifts were analyzed using the Lippert-Mataga approach. In nonpolar hydrocarbon solvents evidence of dual emission from closely spaced (562 cm(-1)) S(1) and S(2) excited states is observed. Preliminary peak assignments for the anomalous S(2) emission are made. PMID- 21648457 TI - Sacrificial template-directed fabrication of superparamagnetic polymer microcontainers for pH-activated controlled release of Daunorubicin. AB - Magnetic pH-sensitive microcontainers were produced by a four-step process. The first step involves the synthesis of citrate-modified magnetic nanoparticles via the coprecipitation method. The second step consists of the encapsulation of magnetic nanoparticles in non-cross-linked poly(methacrylic acid) (PMAA) microspheres through distillation precipitation polymerization, resulting in a core/shell structure. The third step concerns the formation of a poly(N,N' methylenebis(acrylamide)-co-mathacrylic acid) (P(MBAAm-co-MAA)) layer on the surface of magnetic PMAA microspheres by second distillation precipitation polymerization in order to produce a trilayer hybrid microsphere. The last step deals with the removal of PMAA layer in ethanol and formation of a stable P(MBAAm co-MAA) microcontainer with magnetic nanoparticles entrapped inside the formed cavity. This process is simple and leads to the formation of superparamagnetic pH sensitive microcontainers. The structure and properties of the magnetic microcontainers were investigated by X-ray diffraction (XRD), transmission electron microscopy (TEM), scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), vibrating sample magnetometry (VSM), and dynamic light scattering (DLS) to determine the functionalities of the hybrid structure. The magnetic pH-sensitive microcontainers were loaded with Daunorubicin and tested with respect to release rate at different pH values in order to evaluate their functionality as controlled release system. PMID- 21648458 TI - Effect of grain size on uranium(VI) surface complexation kinetics and adsorption additivity. AB - The contribution of variable grain sizes to uranium adsorption/desorption was studied using a sediment from the US DOE Hanford site. The sediment was wet sieved into four size fractions: coarse sand (1-2 mm), medium sand (0.2-1 mm), fine sand (0.053-0.2 mm), and clay/silt fraction (<0.053 mm). For each size fraction and their composite (sediment), batch and flow-cell experiments were performed to determine uranium adsorption isotherms and kinetic uranium adsorption and subsequent desorption. The results showed that uranium adsorption isotherms and adsorption/desorption kinetics were size specific, reflecting the effects of size-specific adsorption site concentration and kinetic rate constants. The larger-size fraction had a larger mass percentage in the sediment but with a smaller adsorption site concentration and generally a slower uranium adsorption/desorption rate. The same equilibrium surface complexation reaction and reaction constant could describe uranium adsorption isotherms for all size fractions and the composite after accounting for the effect of adsorption site concentration. Mass-weighted, linear additivity was observed for both uranium adsorption isotherms and adsorption/desorption kinetics in the composite. One important implication of this study is that grain-size distribution may be used to estimate uranium adsorption site and adsorption/desorption kinetic rates in heterogeneous sediments from a common location. PMID- 21648459 TI - Conformational study of 9-dehydro-9-trifluoromethyl cinchona alkaloids via 19F NMR spectroscopy: emergence of trifluoromethyl moiety as a conformational stabilizer and a probe. AB - The trifluoromethyl substituent has been incorporated into quinidine as a conformational stabilizer and a probe to provide straightforward insight into the conformational behavior of cinchona alkaloids. By significantly decreasing the rotation rate of the quinoline-carbinol bond, the relatively bulky CF(3) group enables the NMR signals of the syn and anti conformers to be differentiated at room temperature. In addition to the stabilizing effect, the introduction of the fluorinated moiety also facilitates the application of (19)F NMR spectroscopy, thereby allowing conformational studies under various conditions without the use of deuterated solvents.